PMID- 10361147 TI - Interaction of a lipid-membrane destabilizing enzyme with PEG-liposomes. AB - Polymer grafted PEG-liposomes have come into use as drug-delivery systems with improved therapeutic profiles. However, very little is known about the morphological instability of PEG-liposomes due to enzymatic degradation. To gain further insight into the effect of PEG lipopolymer-concentration on the catalytic activity of a liposome-degrading enzyme, phospholipase A2 (PLA2)-catalyzed phospholipid hydrolysis of PEG-liposomes has been investigated. The temperature dependence of the PLA2 lag-time, denoting the time required before a sudden increase in enzymatic activity takes place, has been determined for submicellar amounts of dipalmitoylphosphatidylethanolaminyl-poly-(ethylene glycol) (DPPE PEG2000) incorporated into unilamellar dipalmitoylphosphatidylcholine (DPPC) liposomes. The measurements demonstrate a significant reduction in the lag-time over broad temperature ranges. The results suggest that a close relationship exists between PLA2 catalyzed lipid hydrolysis and lipid-membrane composition, which moreover is of major importance for the overall morphological stability and the release of encapsulated material from the polymer-grafted PEG-liposomes. PMID- 10361148 TI - Stabilization of polymer-based gene delivery systems. AB - To preserve the size and transfection potential of polymer-plasmid complexes. Freeze-drying and freeze-thawing were used for stabilization of these complexes. The concentration of the sugars is an important factor affecting both the size and transfection capability of the complexes after freeze-drying and freeze thawing. However, the type of lyoprotectant (sugar) used is of minor importance. It is also shown that when damage to polymer-plasmid complexes occurs, it results from the drying process but is not due to the freezing step. PMID- 10361149 TI - The use of aqueous PEG/dextran phase separation for the preparation of dextran microspheres. AB - A novel procedure to prepare dextran microspheres, without the use of organic solvents was developed. The method is based on phase separation which occurs in aqueous solutions of PEG and methacrylated dextran (dexMA). After stirring this two phase system a water-in-water emulsion is formed. When dexMA forms the discontinuous phase, dextran microspheres can be obtained by polymerization of the methacryloyl groups attached to dextran. The aim of this study was to gain insight into the formulation parameters that affect the particle characteristics. Therefore, it was necessary to establish dexMA/PEG/water phase diagrams. Lower polymer molecular weights and higher degrees of MA substitution resulted in less pronounced phase separation (binodal shifts to higher concentrations). The volume weight mean microsphere diameter varied between 2.5 and 20 microm, depending on the viscosities of both phases and the PEG/dexMA volume ratio. A more viscous continuous phase and/or a less viscous discontinuous phase resulted in smaller microspheres. Furthermore, the particle size increased with decreasing PEG/dexMA volume ratios. The particle characteristics, like cross-link density, initial water content and size can be tailored by adjusting the formulation parameters. PMID- 10361150 TI - Liposomes as cytokine-supplement in tumor cell-based vaccines. AB - Subcutaneous vaccination of C57bl/6 mice with irradiated B16 melanoma cells supplemented with liposomal interleukin-2 (IL2) or murine interferon-gamma (mIFNgamma), resulted in systemic protection in 50% of the animals, against a subsequent tumor cell challenge in a dose dependent manner. The protective efficacy was comparable to the efficacy of cytokine gene-modified cells as tumor vaccine, whereas irradiated B16 cells supplemented with soluble cytokine did not result in protective responses. In vivo evidence was obtained that the beneficial effects mediated by liposome incorporation of the cytokine are the result of a depot function of the liposomal cytokine supplement at the vaccination site. In can be concluded that liposomal delivery of cytokines offers an attractive alternative to cytokine-gene transfection of tumor cells for therapeutic vaccination protocols. PMID- 10361151 TI - Role of macrophages in the localisation of liposomes in lymph nodes after subcutaneous administration. AB - The macrophage 'suicide' technique, based on the ability of clodronate-containing liposomes to deplete lymph nodes of macrophages, was used to study the role of macrophages in lymph node localisation of subcutaneous (s.c.) administered liposomes. Reduced liposome localisation in macrophage depleted lymph nodes confirmed that phagocytosis by macrophages is an important mechanism for lymph node localisation of large (non-sized) liposomes. Depletion of macrophages had less effect on the lymph node localisation of small (about 0.1 microm) liposomes; small liposomes may reach macrophages in regions of lymph nodes not reached by large liposomes. Small liposomes may also be taken up by cells other than macrophages, such as endothelial cells lining the lymph node sinuses. Remarkably, inclusion of poly(ethyleneglycol)-distearoylethanolamine (PEG-PE) into liposomes did not reduce the degree of lymph node localisation in control lymph nodes. As macrophage depletion had a strong negative effect on the lymph node localisation of PEG-liposomes, it is concluded that, despite the presence of a PEG-coating, PEG-liposomes retained by lymph nodes are to a large extent taken up by lymph node macrophages. PMID- 10361152 TI - Characterization of DNA-lipid complexes commonly used for gene delivery. AB - Cationic liposomes are used to deliver genes into cells. Here we describe some poorly understood basic features of DNA-lipid complexes (lipoplexes), especially the electrostatics, stability and DNA structure of lipoplexes, and their effects on transfection (lipofection). Use of the lipophilic, pH-sensitive fluorophore 4 heptadecyl-7-hydroxycoumarin, in combination with Gouy-Chapman calculations, showed that cationic liposomes had a large positive surface potential (180-240 mV) and a high pH (10-11.5) at the location of the probe on the liposomal surface in 20 mM Hepes buffer (pH 7.4). Other electrostatic characteristics were also found, such as the existence of protonable groups of cationic or helper lipids or salt bridges between those. Addition of DNA resulted in neutralization of cationic lipids, which was lower than expected and depending on the type of lipid and the DNA/cationic lipid ratio. The liposomes containing DOTAP (N-(1-(2,3 dioleoyloxy)propyl)-N,N, N-trimethylammonium chloride) were unstable upon dilution, probably due to the high critical micellar concentration of DOTAP, 7x10(-5) M. Large instability expressed as continuous size increase was demonstrated by the time-dependent static changes in light-scattering monitored following the mixing of cationic liposomes and DNA at DNA/cationic lipid molar ratios between 0.2 and 0.8. Addition of cationic liposomes composed of 100% DOTAP or DOTAP/DOPE (1/1) liposomes, induced instantaneous transition of the plasmid DNA from the B- toward a partial C-type conformation as shown by circular dichroism (CD) spectroscopy and at certain conditions Psi--DNA could be found as well. The Psi--DNA is characterized by inter-helical interaction between parallel helices. The highest lipofection was obtained under conditions of lipoplex instability, and when the DNA was partially dehydrated and had a partial Psi-- structure. PMID- 10361153 TI - Inverse toroidal vesicles: precursors of tubules in sorbitan monostearate organogels. AB - Sorbitan monostearate organogels are opaque, thermoreversible semi-solids whose microstructure consists of surfactant tubules dispersed in the organic continuous phase. Inverse toroidal vesicles are the precursors of the surfactant tubules. The gelation process was observed as an isotropic sol phase of sorbitan monostearate in isopropyl myristate was cooled using hot-stage light microscopy. At the gelation temperature, inverse toroidal vesicular structures were seen to grow in the organic phase. These toroids are thought to be analogous to other well-known vesicles, liposomes and niosomes, except for their toroidal (rather than spherical) shape and their inverse nature. They are rather short-lived structures: on further cooling of the sol phase, tubules form in the organic medium: it is speculated that the toroids elongate into tubular shapes or split into rod-shaped segments. PMID- 10361154 TI - Distribution of a lipidic 2.5 nm diameter dendrimer carrier after oral administration. AB - The biodistribution of a lipidic peptide dendrimer has been studied after oral administration to female Sprague-Dawley rats (180 g, 9 weeks old). Uptake by gut epithelial tissue of the radiolabelled dendrimer molecule (mol. wt. 6300; diameter 2.5 nm; log P=1.24) was studied in rats after a single oral dose by gavage (14 mg/kg). The maximum levels of dendrimer observed were 3% (blood), 1.5% (liver), 0.1% (spleen), 0.5% (kidneys), 15% (small intestine) and 5% (large intestine). Approximately 6% of a single administered dose (28 mg/kg) was recovered from the entire gastrointestinal tract while 1% was absorbed via the small intestine lymphoid tissue after 3 h; after 12 h, 0.1% was detected. The maximum uptake by the non-lymphoid small intestine was 4% of the dose after 3 h. After 12 h, 0.3 and 4% dendrimer was measured in the lymphoid large intestine and the non-lymphoid large intestine, respectively. The total percentage of the administered dose absorbed through the lymphoid tissue was comparatively greater than through the non-lymphoid tissue of the small intestine with respect to organ weight after 3 and 24 h. PMID- 10361155 TI - In vitro/in vivo characterisation of polyhedral niosomes. AB - Non-ionic surfactant vesicles (niosomes) formed by a hexadecyl diglycerol ether (C16G2) and a series of polyoxyethylene alkyl ethers exhibit a variety of shapes dependent on their membrane composition. These surfactants form with an equimolar amount of cholesterol a mixture of largely spherical and tubular niosomes. In the absence of cholesterol, they form faceted polyhedral structures. The physicochemical and biological differences between polyhedral and spherical/tubular niosomes were studied. Polyhedral niosomes undergo a reversible shape transformation into spherical structures on heating above their phase transition temperature (Tm). The viscosity of polyhedral niosomes at room temperature is higher than their spherical counterparts due to their faceted and relatively rigid shape, and is more dependent on temperature due to shape transformation. At room temperature, polyhedral niosomes possess more rigid gel phase membranes and are less osmotically sensitive; however, they are more permeable because of a lack of or low levels of cholesterol in their membranes. Polyhedral niosomes loaded with luteinising hormone releasing hormone (LHRH), nonetheless, slow the release of drug compared to solution, albeit to a small extent. PMID- 10361156 TI - Physico-chemical characterization of insulin-loaded poly(isobutylcyanoacrylate) nanocapsules obtained by interfacial polymerization. AB - Insulin could be encapsulated very efficiently in oily containing poly(isobutylcyanoacrylate) nanocapsules obtained by interfacial polymerization. In addition, these nanocapsules showed unexpected biological activity after intragastric administration. The hypoglycemic effect was characterized by a lag time period of 2 days and a prolonged effect over a period of 20 days. To explain, the high encapsulation rate of insulin achieved in these nanocapsules and the biological effect, this work was focused on the characterization of the nanocapsules and on the study of the mechanism of nanocapsule formation. Results showed that insulin was found unmodified during the nanoencapsulation process. This was due to the large amount of ethanol used in the preparation of the nanocapsules that initiated the polymerization of isobutylcyanoacrylate preserving the peptide from a reaction with the monomer. Results also showed that insulin was located inside the core of the nanocapsules and not simply adsorbed onto their surface. PMID- 10361157 TI - Optimization of the encapsulation and release of beta-lactoglobulin entrapped poly(DL-lactide-co-glycolide) microspheres. AB - The goal of the present paper was to optimize the encapsulation of beta lactoglobulin (BLG) within poly(lactide-co-glycolide) (PLGA) microparticles prepared by the multiple emulsion solvent evaporation method. The role of the pH of the external phase and the introduction of the surfactant Tween 20, in the modulation of the entrapment and release of BLG from microparticles, was studied. Reducing the solubility of BLG by decreasing the pH of the external phase to a value close to the pI of BLG resulted in a better encapsulation with, however, a larger burst release effect. By contrast, Tween 20 was shown to increase the encapsulation efficiency of BLG and reduce considerably the burst release effect. In fact, Tween 20 was shown to be responsible for removing the BLG molecules that were adsorbed on the particle surface. In addition, Tween 20 reduced the number of aqueous channels between the internal aqueous droplets as well as those communicating with the external medium. Thus, the more dense structure of BLG microspheres could explain the decrease in the burst release. These results constitute a step ahead in the improvement of an existing technology in controlling protein encapsulation and delivery from microspheres prepared by the multiple emulsion solvent evaporation method. PMID- 10361158 TI - In vivo fate and immune pulmonary response after nasal administration of microspheres loaded with phosphorylcholine-thyroglobulin. AB - Phosphorylcholine is a widely occurring hapten which is present in the cell wall of many prokaryotes. It is, therefore, an attractive candidate for the development of a vaccine against many bacterial diseases. Poly(D,L-lactide-co glycolide) microspheres loaded with phosphorylcholine linked to thyroglobulin (PC Thyr) as protein carrier were prepared. The effect of the protein concentration on antigen encapsulation and release as well as on microsphere morphology has been investigated. When administered intranasally, PC-Thyr-loaded microspheres were taken up by epithelial cells of the nasopharyngeal associated lymphoid tissue and induced a specific IgA and IgG response in pulmonary secretions as well as a strong systemic immune response in BALB/c mice. PMID- 10361159 TI - Localized paclitaxel delivery. AB - While the search for new antineoplastic agents is in progress, optimization of delivery for existing drugs will remarkably improve the current scenario in the management of cancer. Paclitaxel, a new antineoplastic agent, is one such drug deserving attention in the field of regional drug delivery, offering immense pharmacokinetic as well as therapeutic advantage via localized delivery. The antiangiogenic activity of paclitaxel has been demonstrated using the chick chorioallantoic membrane model (CAM). This review focuses on the antiangiogenic activity of paclitaxel supported by the evidence that angiogenesis inhibitors display potential synergism with cytotoxic agents in the treatment of primary and metastatic cancers. Preclinical trials have confirmed that the biological and cytotoxic effects of paclitaxel on several tumor cell lines are enhanced by the increase in both the drug concentration and the duration of exposure. Sufficient experimental evidence has accumulated to state that localized delivery will exploit the multiple pharmacological effects of paclitaxel in the treatment of refractory and metastatic cancerous diseases. The drug delivery systems, namely, microspheres, surgical pastes and implants, fabricated for localized paclitaxel delivery are reviewed explaining the concept of increased tumor burden alleviating body burden as a consequence of such delivery systems. Some of the preclinical trials are very encouraging and speculate a promising future for these devices in the battle against solid tumors. Finally, the review briefs on the possibilities for better paclitaxel delivery and the future drug delivery systems for localized cancer chemotherapy. PMID- 10361160 TI - Vesicle formation from hexasubstituted cyclophosphazenic derivatives. AB - Hexakis[butoxytris(ethoxy)]cyclophosphazene (3a), hexakis[dodecyloxytetrakis (ethoxy)]cyclophosphazene (3b) and hexakis[hexadecyloxyeicosanekis(ethoxy)]cyclophosphazene+ ++ (3c) were synthesised and their ability to form niosomes was studied. All synthesised compounds in the presence of cholesterol were shown to form vesicles, which aggregated strongly. To prevent aggregation, dicetylphosphate was used. The capacity of the sonicated and unsonicated niosomes to encapsulate hydrophile and lipophile molecules was also studied using carboxyfluorescein and diphenylhexatriene. PMID- 10361161 TI - Photodegradation of amiloride in aqueous solution. AB - The photodegradation of amiloride hydrochloride in deaerated aqueous solution at 30 degrees C in the pH range 4.5-11.0 was studied by spectrophotometry and reversed-phase HPLC. The neutral form of the drug present in alkaline solution degraded approximately 3-fold faster than the cationic form. The quantum yields for photodegradation of neutral amiloride and its conjugate acid were determined using ferrioxalate actinometry to be 0.023+/-0.002 and 0. 009+/-0.001, respectively. The initial photoreaction involves dechlorination of amiloride and the major product is N-amidino-3, 5-diamino-6-hydroxylpyrazine-carboxamide, established by UV, 1H and 13C NMR, and chemical ionization-mass spectrometry. The mechanism of photolysis is postulated to involve cation radical formation that facilitates the dechlorination step. The photosensitizing activity of amiloride hydrochloride was tested by measuring (a) the rate of oxygen uptake in the presence of singlet oxygen substrates, 2, 5-dimethylfuran or l-histidine, and (b) the rate of free radical polymerization of acrylamide, at 30 degrees C in aqueous solution. Photosensitization by amiloride was concluded to occur predominantly via singlet oxygen rather than a free radical mechanism. However, amiloride is a much weaker photosensitizer than other diuretics such as frusemide and hydrochlorothiazide, tested under the same experimental conditions. PMID- 10361162 TI - Transdermally delivered peroxovanadium can lower blood glucose levels in diabetic rats. AB - The element vanadium can have insulin mimetic properties and therefore has been suggested as a possible therapeutic agent for treatment of diabetes. A series of peroxovanadium compounds that are more potent at lowering blood glucose levels than sodium metavanadate, sodium orthovanadate and vanadyl sulfate have recently been synthesized. These compounds probably will not be orally active so transdermal administration is a potential option. A patch containing either the peroxovanadium compound [VO(O2)2 1-10 phenanthroline], abbreviated bpV(phen), or placebo was placed on the back of streptozotocin induced diabetic rats and was delivered either passively (16 h) or iontophoretically (0.5 mA/cm2 for 4 h). Blood samples were analyzed for glucose and vanadium levels. Mean blood glucose levels were 83+/-1% and 109+/-1% of the starting values for animals iontophoretically treated with bpV(phen) and vehicle, respectively. The compound's insulin mimetic properties were evident within 60 min of current initiation. Blood glucose levels were reduced to 74+/-14% of the original level after 16 h of passive treatment. The compound was ineffective when fed to animals. Transdermal delivery of bpV(phen) resulted in significantly greater blood levels of vanadium than the orally delivered compound (P<0.05). Overall these experiments demonstrate that peroxovanadium delivered through the skin can lower blood glucose levels in rats. Further experiments are warranted to better characterize the nature of the response and to determine the potential for using these compounds in humans. PMID- 10361163 TI - Enhanced rectal absorption of insulin-loaded Pluronic F-127 gels containing unsaturated fatty acids. AB - The objective of this study was to prepare and to evaluate Pluronic F-127 (PF127) gel containing unsaturated fatty acids such as oleic acid (18:1), eicosapentaenoic acid (20:5) and docosahexaenoic acid (22:6) as a potential formulation for rectal delivery of insulin. The hypoglycemic effect of insulin was examined following rectal administration of the various formulations in normal rats. Rectal insulin absorption was markedly enhanced, and marked hypoglycemia was induced by all PF127 gels (insulin dose, 5 U/kg) containing different unsaturated fatty acids. PF127 gels containing unsaturated fatty acids presented low tmax mean values indicating that the absorption of insulin occurred very rapidly in the rectum. The relative hypoglycemic efficacy of PF127 gel formulations containing fatty acids such as oleic acid, eicosapentaenoic (EPA) and docosahexaenoic (DHA) were 28.4+/-8.1, 26.8+/-14.3 and 23.1+/-5.7%, respectively. The finding demonstrated that 20% PF127 gels containing unsaturated fatty acids are potential formulations for rectal delivery of insulin. PMID- 10361164 TI - Description and preliminary evaluation of a new ultrasonic atomizer for spray congealing processes. AB - A new atomizer that operates with ultrasonic energy is described. This apparatus is intended to obtain microparticulate drug delivery systems through spray congealing or spray-drying technologies. In this work, some experimental results are reported on model systems submitted to spray-congealing. The formulations under examination contained theophylline and fenbufen as model drugs and stearic acid, carnauba wax, Cutina HR(R) and Compritol 888 ATO(R) as low melting excipients. Non-aggregate and spherical-shaped microparticles were obtained with all the materials tested; moreover, they had smooth surface and good flowability. The particle sizes depend on the amount of drug present and in each case the maximum size value of the distribution frequency was found to be 375 mu. In vitro release of the drug depends on its solubility and on the excipient lipophilicity. The results suggest that the ultrasound-assisted atomizer could be proposed as a possible alternative to traditional atomizers used for spray-congealing in the pharmaceutical field. PMID- 10361165 TI - Phospholipid-based microemulsions of flurbiprofen by the spontaneous emulsification process. AB - The purpose of this study was to investigate the possibility for parenteral delivery of flurbiprofen without chemical modification using a phospholipid-based microemulsion system. Microemulsions composed of ethyl oleate, lecithin and distearoylphosphatidyl-ethanolamine-N-poly(ethyleneglycol) 2000 (DSPE-PEG) were prepared using ethanol as a cosolvent. The effect of formulation variables on the particle size of the microemulsion was investigated. Flurbiprofen concentrations in plasma and various organs after the intravenous administration of flurbiprofen loaded microemulsion were measured and compared with those after the intravenous administration of flurbiprofen axetil-entrapped emulsion (Lipfen(R), 50 mg/5 ml as flurbiprofen axetil) and flurbiprofen solution. Phospholipid-based microemulsions could solubilize more than 10 mg ml-1 of flurbiprofen at the ratio of vehicle to drug at least 10:1, if the oil contents (10 or 20%) of common parenteral emulsions were used. The half-life, AUC and MRT of flurbiprofen loaded in microemulsion (ethyl oleate:lecithin:DSPE-PEG:flurbiprofen=8:3:1:1.2) increased significantly. The biodistribution of flurbiprofen loaded in this microemulsion was quite different from others. Reticuloendothelial uptake of flurbiprofen loaded in microemulsion decreased compared with that in solution or Lipfen(R). It is concluded that the current microemulsion system might be applicable to formulate the parenteral dosage form of poorly water-soluble flurbiprofen without chemical modification. PMID- 10361166 TI - Evaluation of melt agglomeration properties of polyethylene glycols using a mixer torque rheometer. AB - Lactose was melt agglomerated in a mixer torque rheometer with polyethylene glycol (PEG) 2000, 3000, 6000, 8000, 10 000, or 20 000 as meltable binder. A longer massing time caused an increase in mean torque until a maximum value after which the torque decreased. A smaller particle size of the PEG gave rise to a faster initial rise in mean torque. The higher viscosity of the PEG 20 000 resulted in a higher mean torque, whereas no clear difference in mean torque was obtained with the other PEGs. The binder concentration could be varied within a rather wide range without causing overwetting, the range being wider with PEG 3000 than with PEG 20 000. The mean torque values obtained were found to be related to the liquid saturation of the agglomerates. The reproducibility of the experiments was found to be very dependent on the experimental conditions, the highest binder viscosities and binder concentrations giving rise to a poor reproducibility. The results were compared with a few melt agglomeration experiments with PEG 3000 in a high shear mixer. The mixer torque rheometer was found not to be suitable for predicting melt agglomeration properties in the high shear mixer because of a marked difference in the shear forces in the two mixers. PMID- 10361167 TI - Influence of the level of ceramides on the permeability of stratum corneum lipid liposomes caused by a C12-betaine/sodium dodecyl sulfate mixture. AB - The sublytic interactions of a mixture of N-dodecyl-N, N-dimethylbetaine dodecyl betaine (C12-Bet)/sodium dodecyl sulfate (SDS) (mole fraction of the zwitterionic surfactant=0.6) with stratum corneum (SC) lipid liposomes varying the proportion of ceramides type III (Cer) were investigated. The surfactant/lipid molar ratios (Re) and the bilayer/aqueous phase partition coefficients (K) were determined by monitoring the changes in the fluorescence intensity of liposomes due to the 5(6) carboxyfluorescein (CF) released from the interior of vesicles. The fact that the free surfactant mixture concentration was always lower than its critical micelle concentration indicates that permeability changes were ruled by the action of surfactant monomers in all cases. Higher and lower Cer proportions than that of the SC lipids led to a fall and to a rise in the activity of the surfactant mixture on these bilayer structures. However, the surfactant partitioning into liposomes (or affinity with these bilayer structures) increased as the proportion of Cer increased up to the highest value was achieved for a Cer proportion similar to that in the SC lipids (about 40-45%). Thus, at low Cer proportions the ability of the surfactant mixture to alter the permeability of these bilayer structures was higher than that for liposomes approximating the SC lipid composition despite their reduced partitioning into liposomes. These findings are in agreement with the recently reported dependencies of the level of ceramides in skin lipids and function barrier abnormalities and could explain in part these dependencies. PMID- 10361168 TI - Physicochemical properties of chitosan-lipid emulsions and their stability during the autoclaving process. AB - A new positively charged, submicronized fat emulsion with appropriate stability during the autoclaving process was developed. Only the emulsions prepared with a combination of ABA block co-polymer (F68) and chitosan were stable enough to resist the thermic shock induced by autoclaving sterilization. The results indicate that a mixed film consisting of the ABA block co-polymer and chitosan molecules was formed at the o/w interface with an overall positive surface charge. Conversely, a combination between chitosan with phospholipids and/or with a mixture of phospholipids with ABA block co-polymer showed a phase separation during autoclaving. A chitosan type with a low viscosity was used which was intended for a possible use in the ocular and parenteral application. An experimental factorial design 32 was used to investigate the effect of chitosan and F68 concentrations on the physicochemical properties of the system and consequently their influence on the stability of emulsions during autoclaving. Both size and surface charge of emulsions were significantly affected as a function of the chitosan concentration. Formulation with a mean particle size ranging from 125 to 130 nm and with a positive surface charge of 20-23 mV was achieved. Moreover, the chitosan emulsions were autoclaved without a significant change in their particle size. However, increasing the concentration of chitosan needs a higher amount of F68 in order to achieve stable emulsions during autoclaving. This may be due to the interaction between the positively-charged chitosan and the negatively-charged free fatty acids, which are contained in the oil phase (castor oil). PMID- 10361169 TI - Development of dried liposomes containing beta-galactosidase for the digestion of lactose in milk. AB - The hydrolyzed-lactose milk for lactase-deficient subjects has a sweeter taste than whole milk, and some subjects dislike its taste. In order to cope with this shortcoming, we examined whether beta-galactosidase, which hydrolyzes lactose, added to the whole milk in the form of dried liposomes, would be able to digest lactose in milk following the lysis of liposomes in the presence of bile salts. Dried liposomes containing beta-galactosidase were prepared in the presence of trehalose by the dehydration-rehydration vesicle method to overcome the instability of the conventional liposome suspension. The stability of liposomal membranes was evaluated by measuring the activity of entrapped beta-galactosidase under various storage conditions. By treating liposomes with trehalose, which was found to prevent the fusion of liposomes and the leakage of entrapped drug, the entrapping efficiency increased up to fourfold. Over 95% of dried liposomes which had been stored at 17 degrees C for 60 days were reconstituted to liposomes upon rehydration process. From the stability study, dried liposomes were found to retain 87% of beta-galactosidase activity at 17 degrees C after 60 days and to be more stable than the multilamellar vesicle suspension prepared without trehalose. The lysis study showed that dried liposomes were hardly lyzed in the simulated gastric fluid with pepsin, but lyzed immediately more than 90% in 0.01 M deoxycholic acid. Lactose hydrolysis in the presence of deoxycholic acid after the addition of dried liposome-entrapped beta-galactosidase to whole milk was proportional to the quantity of entrapped beta-galactosidase and the amount of dried liposomes added. These results demonstrate that beta-galactosidase entrapped in liposome is stable and reconstituted mostly upon rehydration, and can digest lactose in milk after the efficient lysis of liposomes in the presence of bile salts. This study implies that beta-galactosidase entrapped in liposome may be applied to whole milk for lactase-deficient subjects. PMID- 10361170 TI - Physico-chemical characterisation and transfection efficiency of lipid-based gene delivery complexes. AB - Cationic liposomes spontaneously interact with negatively charged plasmid DNA to form a transfection competent complex capable of promoting the expression of a therapeutic gene. This work aims to improve the understanding of the poorly defined mechanisms and structural rearrangements associated with the lipid-DNA interaction. Specifically, dimethyl dioctadecylammonium bromide (DDAB):dioleoyl phosphatidylethanolamine (DOPE) and 1,2-dioleoyl-3-trimethylammonium propane (DOTAP) liposomes were mixed with a reporter plasmid (pADbeta or pCMVbeta) to form lipid-DNA complexes. The size and charge characteristics of the complexes as determined by photon correlation spectroscopy and microelectrophoresis were found to be dependent on the lipid:DNA ratio, with both DDAB:DOPE-DNA and DOTAP-DNA complexes aggregating at around neutral zeta potential. Negative stain transmission electron microscopy demonstrated at least three distinct complex structures being formed at the same DOTAP:DNA ratio. We postulate that two of these aggregates are structural moieties involved in the formation of the efficient transfection particle. Gel electrophoresis was used to determine the efficiency and extent of lipid-DNA complex formation. Results showed that only DOTAP liposomes were capable of preventing ethidium bromide intercalation with DNA and protecting the enclosed plasmid from nuclease digestion. When a range of lipid-DNA complexes were transfected into in vitro cell lines, the efficiency of reporter gene (beta-galactosidase) expression was found to depend on the type of liposome used in the complex, the ratio of lipid:DNA and the transfected cell line. Our results challenge the requirement for DOPE to be included in the formulation of cationic lipid vectors, especially in the case of DOTAP containing liposomes. PMID- 10361171 TI - Inguinal hernia repair. PMID- 10361172 TI - Obscure lower gastrointestinal tract bleeding. PMID- 10361173 TI - Endoleak after stent-graft treatment of abdominal aortic aneurysm: a meta analysis of clinical studies. AB - BACKGROUND: Endoleak is the major complication after endovascular treatment of abdominal aortic aneurysm (AAA) and its incidence seems to remain significant. Little is known about the association of device type and configuration with respect to the incidence, location, time of onset and fate of endoleakage. METHODS: A meta-analysis was performed via a Medline search of clinical studies after 1995 dealing with the endovascular treatment of AAA. Details of number of patients treated, configuration and type of endovascular device were collected. Data concerning site of origin, time of occurrence and fate of the endoleak were retrieved, along with information on change in diameter of the aneurysm with time. RESULTS: The 23 publications included reported on 1189 patients. The 1118 patients with successfully inserted transfemoral endovascular grafts experienced 270 endoleaks (24 per cent). The majority arose from the distal stent attachment site (36 per cent), were present immediately after stent-graft placement (66 per cent) and were persistent in time (37 per cent). Tube grafts were more frequently affected by endoleakage (35 per cent; P < 0.0001), especially at the distal stent attachment site (51 per cent), than bifurcated grafts (18 per cent; P = 0.004) and aortounilateral devices (20 per cent; P = 0.70). Self- expandable stent grafts were more frequently associated with endoleaks (25 per cent) than balloon expandable stent-grafts (17 per cent) (P = 0.037). CONCLUSION: Endovascular treatment of AAA is an evolving field. Even after the initial learning curve and attention to device-related problems, it is still accompanied by a significant number of endoleaks. Uniform presentation of results of treatment is necessary for analysing the effect of differences between patients, aneurysm morphology and device type. PMID- 10361174 TI - Periampullary diverticula and pancreaticobiliary disease. AB - BACKGROUND: Periampullary diverticula (PAD) are extraluminal outpouchings of the duodenum arising within a radius of 2-3 cm from the ampulla of Vater. They are frequently encountered in elderly patients undergoing endoscopic retrograde cholangiopancreatography (ERCP) and contribute to failure of ERCP. This review details the relationship of PAD to pancreaticobiliary disease and the influence of PAD on the management of patients. METHODS: The United States National Library of Medicine Medline database was searched for articles on and related to PAD published in English within the past 15 years. Major earlier works were also reviewed. RESULTS: The prevalence of PAD increases with age and could be as high as 27 per cent. PAD are associated with an incompetent sphincter of Oddi and colonization of bile duct with beta-glucuronidase-producing organisms. PAD are implicated in the pathogenesis of pigment common bile duct stones, but there is no conclusive evidence to associate them with cholecystolithiasis or pancreatitis. PAD are a major cause of failure of ERCP, but success rates of more than 90 per cent have been achieved in specialist centres. CONCLUSION: With an ageing population, there will be an increase in elderly patients with PAD and symptomatic pancreaticobiliary disease. Continuing improvements in radiological and endoscopic techniques should enable this vulnerable group to be treated effectively and safely. PMID- 10361176 TI - Digest PMID- 10361175 TI - Two articles for comparison. Article A: APACHE II score in massive upper gastrointestinal haemorrhage from peptic ulcer: prognostic value and potential clinical applications. Article B: APACHE II score in massive upper gastrointestinal haemorrhage from peptic ulcer. PMID- 10361177 TI - Prospective randomized comparison between pylorus-preserving and standard pancreaticoduodenectomy. AB - BACKGROUND: Pancreaticoduodenectomy, with either gastrectomy (Whipple procedure) or pylorus-preserving pancreaticoduodenectomy (PPPD), is a complex procedure. Technical diversity, variation and sampling bias exist among surgeons. Previous reports comparing these two procedures are retrospective and not randomized. These factors should be considered seriously and eliminated in comparisons between the two procedures. METHODS: From August 1994 to August 1997, a prospective randomized comparison was conducted between the Whipple procedure and PPPD performed by the same surgeon with the same approach and same anastomotic fashion for periampullary cancer. After exclusion of seven patients, 31 patients were eligible for the study, 16 receiving PPPD and 15 a Whipple procedure. No significant difference in the age, sex distribution, tumour localization or staging was noted between the two groups. RESULTS: One operative death after PPPD and no operative death after the Whipple procedure resulted in a 3 per cent mortality rate in the 31 patients. Median duration of the Whipple operation was 235 (range 195-305) min, with a median blood loss of 500 (range 230-3100) ml and a median blood transfusion of 0 (range 0-10) units. In the patients who had PPPD, median operating time was 230 (range 170-275) min, median blood loss was 350 (range 100-1200) ml and median blood transfusion was 0 (range 0-4) units. There were two minor leaks from the pancreaticojejunostomy after the Whipple procedure and no leakage after PPPD, resulting in 6 per cent minor leakage in 31 patients. These outcomes were not significantly different. Delayed gastric emptying was observed more frequently after PPPD (six of 16 patients) than after the Whipple procedure (one of 15 patients), with marginal significance (P = 0.08, two-sided Fisher's exact test). CONCLUSION: In this prospective randomized study, both PPPD and the Whipple procedure were associated with low mortality and operative morbidity rates. There was no significant difference between PPPD and Whipple resection in terms of operative mortality and morbidity, operating time, blood loss and blood transfusion. PPPD was associated with more frequent delayed gastric emptying, although study of more patients is needed to confirm this. PMID- 10361178 TI - Randomized clinical trial of ultrasonic dissector or conventional division in distal pancreatectomy for non-fibrotic pancreas. AB - BACKGROUND: Resection of the non-fibrotic pancreas is prone to postoperative pancreatic fistula because of well preserved exocrine secretions and easily crushed soft parenchyma. The purpose of this study was to evaluate ultrasonic dissection for division of the non-fibrotic pancreas in distal pancreatectomy. METHODS: All pancreata included in this study were soft on direct palpation and their main ducts had no dilatation, at least proximally from the transection line. Fifty-eight patients with gastric cancer or pancreatic disease were randomly assigned to the two groups. In the ultrasonic dissection (UD) group (n = 27), all pancreatic ducts were identified and ligated securely. The stump was left open without parenchymal suturing. In the conventional (CV) group (n = 31), the pancreas was cut with a knife and the stump was oversewn in mattress fashion. The main pancreatic duct was ligated in all patients in both groups. Pancreatic fistula was defined as a pancreatic fluid discharge for more than 7 days after operation diagnosed according to amylase concentration in the drainage fluid. RESULTS: In the UD group, approximately 20-30 tubes including a mean(s.d.) 5.2(0.8) (range 4-6) pancreatic ducts were skeletonized and ligated per patient. There were nine pancreatic fistulas (16 per cent); one in the UD group and eight in the CV group (P = 0.020). CONCLUSION: In distal pancreatectomy for the non fibrotic pancreas, ultrasonic dissection without suture closure of the stump reduced the incidence of pancreatic fistula compared with conventional division and suture, in this randomized trial. PMID- 10361179 TI - Randomized trial of open versus closed day-case haemorrhoidectomy. PMID- 10361180 TI - Randomized trial to study the effect of fundic mobilization on long-term results of Nissen fundoplication. AB - BACKGROUND: It has been suggested that fundic mobilization in Nissen fundoplication decreases adverse postoperative symptoms and increases the durability of the fundic wrap. There are no previous randomized, prospection long term studies assessing this. This study addresses the question. METHODS: Fifty consecutive patients were randomized to undergo open Nissen-Rossetti fundoplication with total fundic mobilization (n == 26) or an identical procedure without mobilization (n == 24). After a median of 36 (range 6-53) months 49 had endoscopy and a personal interview. RESULTS: Oesophagitis was cured in 20 of 21 patients who had fundic mobilization and 18 of 19 who did not. a defective fundic wrap was observed in five and recurrent reflux symptoms in six of those who had fundic mobilization compared with two and one respectively of those who did not (P not significant). Recurrent sliding hiatal hernia was observed in nine of those with a mobilized fundus and one of those without (P == 0.02). There was no significant difference in incidence of new-onset long-term dysphagia, subjective belching ability, amount of flatus or bloating between the study groups. CONCLUSION: Fundic mobilization did not give any apparent advantage regarding postoperative adverse effects. Instead, it was associated with a higher rate of recurrent hiatal hernia. PMID- 10361181 TI - Follow-up of patients with a thin melanoma. AB - BACKGROUND: The aim of this study was to determine the value of follow-up in two subgroups of patients with a thin melanoma less than 0.76 mm and 0.76-1.5 mm thick. METHODS: The study group comprised all patients presenting to the Cardiff Melanoma Clinic from its introduction in 1976 to the end of 1994. All patients attend follow-up according to a strict protocol, determined by the thickness of the original melanoma (less than 0.76 mm, annually; more than 0.76 mm, every 2 months for 2 years, 3 monthly for 2 years, 4 monthly for 1 year and then annually). RESULTS: In total there were 306 patients with a thin melanoma: 178 with a melanoma thinner than 0.76 mm (group 1) and 128 with a melanoma of 0.76 1.5 mm (group 2). The groups were well matched for age (mean 50.6 (range 8-87) versus 50.6 (range 19-89) years respectively) and length of follow-up (mean 88.6 (range 4-296) versus 80.4 (range 2-296) months). Four patients (2.2 per cent) developed recurrence in group 1 and 16 (12.5 per cent) in group 2. The mean time to recurrence was 84.5 (range 49-143) months in group 1 and 45.3 (range 2-74) months in group 2. All patients in group 1 and 14 of 16 in group 2 died from recurrent disease. CONCLUSION: Follow-up of patients with a melanoma less than 0.76 mm thick is not worthwhile. All recurrences would have been detected by annual review for 7 years in patients with melanomas between 0.76 and 1.5 mm thick. PMID- 10361182 TI - Treatment of gallbladder cancer by radical resection. AB - BACKGROUND: The use of radical resection for gallbladder cancer is controversial. This study evaluated results of resection for gallbladder cancer and analysed prognostic factors. METHODS: A retrospective review of 135 patients who underwent surgical resection for gallbladder cancer between 1976 and 1998 was performed. Of these, 123 patients underwent radical resection and the remaining 12 had palliative resection. The resections included 32 hepatopancreatoduodenectomies and 57 with adjuvant radiotherapy. Twelve prognostic factors were analysed. A subset of 96 patients with stage IV disease was analysed separately with respect to residual tumour level and adjuvant radiotherapy. RESULTS: Surgical resection was associated with a 5-year survival rate of 36 per cent, with a mean follow-up time of 870 days. Twenty-two patients have survived more than 5 years including three with stage IV disease. Overall operative morbidity and mortality rates were 13 and 4 per cent respectively. The 5-year survival rate decreased with disease stage: 100, 78, 69 and 11 per cent for stages I (n = 13), II (n = 19), III (n = 7) and IV (n = 96) respectively. Performance status, jaundice, histopathological type and grade, primary tumour, lymph node, distant metastasis, stage grouping, residual tumour level and adjuvant radiotherapy were significant prognostic factors. CONCLUSION: With careful patient selection, radical resection for gallbladder cancer improves the prognosis with acceptable operative mortality and morbidity rates, even for stage IV disease, provided that complete gross tumour resection is combined with radiotherapy. PMID- 10361183 TI - Characterization of the Kupffer cell response to exogenous endotoxin in a rodent model of obstructive jaundice. AB - BACKGROUND: Sepsis and endotoxaemia occur frequently in biliary obstruction. Impaired Kupffer cell endocytosis is implicated in these events. Tumour necrosis factor and interleukin 6, secreted by Kupffer cells, are important mediators of sepsis. Kupffer cell clearance of endotoxin and secretion of cytokines in experimental obstructive jaundice were investigated. METHODS: Wistar rats were randomized to bile duct ligation, sham operation or control. Groups (n = 8) were studied 1 and 3 weeks after operation. Kupffer cell function was assessed using in situ hepatic perfusion. RESULTS: Clearance of endotoxin was significantly depressed 1 week (median (interquartile range) 20.3 (10.5-27.1) per cent) and 3 weeks (22.1 (20.2-23.2) per cent) after bile duct ligation compared with that in respective sham animals (35.5 (29.9-41.6) and 40.9 (37.7-47.0) per cent) and controls (39.5 (37.3-46.8) per cent). Secretion of tumour necrosis factor was significantly greater 1 week (1113.7 (706.5-1436. 8) pg/ml) and 3 weeks (1118.2 (775.7-1484.1) pg/ml) following bile duct ligation compared with that in respective sham animals (114.3 (0-178.5) and 107.6 (63.7-166.4) pg/ml) and controls (0 (0-20.7) pg/ml). Interleukin 6 was not secreted by sham or control animals but was present in the perfusate from jaundiced animals at 1 and 3 weeks (52.5 (9.9-89.5) and 66.2 (60.2-193.1) pg/ml). CONCLUSION: These data demonstrate simultaneous impairment of Kupffer cell clearance of endotoxin and increased secretion of proinflammatory cytokines in experimental obstructive jaundice. These diverse responses may contribute to the development of sepsis-related complications in biliary obstruction. PMID- 10361184 TI - Immunoglobulin A antibodies against Chlamydia pneumoniae are associated with expansion of abdominal aortic aneurysm. AB - BACKGROUND: The aim of this study was to examine the possible association between the progression of small abdominal aortic aneurysm (AAA) and chronic infection with Chlamydia pneumoniae. METHODS: Patients from a hospital-based mass screening programme for AAA with annual follow-up (mean 2.7 years) were included. After initial interview, 139 men aged 65-73 years with a small AAA underwent examination and blood sampling. Immunoglobulin (Ig) G and IgA titres against C. pneumoniae were measured by a microimmunofluorescence test. RESULTS: Some 83 (95 per cent confidence interval 74-93) per cent of the men had an IgA titre of 20 or more, or an IgG titre of 32 or more. Men with an IgA titre of 20 or more had a 48 per cent higher AAA expansion rate than those with a titre of less than 20 (3.1 versus 2.1 mm/year; P < 0.05). Multiple linear and logistic regression analyses showed that an IgA titre of 20 or more was a significant independent predictor of increased AAA expansion, adjusted for known risk factors of expansion. Initial AAA size and serum total cholesterol level were also predictors of expansion. CONCLUSION: A high proportion of men with a small AAA had signs of chronic infection with C. pneumoniae. Aneurysm progression correlated with evidence of chronic C. pneumoniae infection. PMID- 10361185 TI - Contribution of diet, tumour volume and patient-related factors to weight loss in patients with colorectal liver metastases. AB - BACKGROUND: One of the difficulties in assessing the contribution of tumour related factors to cancer cachexia is measurement of the extent of disease where dissemination to multiple organ sites has occurred. METHODS: In this study the extent of tumour (both tumour volume and increase in marker levels), diet and patient-related factors (appetite, metabolic hormones, immune activation, liver function and quality of life) were compared in patients with colorectal liver metastases who had lost at least 1 kg in body-weight (weight loss) and patients who had not lost 1 kg in body weight (stable weight) during the preceding month. RESULTS: Forty patients (22 men; 14 with weight loss) were studied. Liver metastasis volume was significantly greater in patients who lost weight than in those whose weight was stable (median (interquartile range) 1179 (245-1517) versus 119 (23-523) ml; P = 0.003). The prevalence of patients with raised levels of serum immune products was significantly greater in the weight loss group for soluble interleukin (IL) 2 receptor alpha (sIL2ralpha) (P = 0.03) and IL-6 (P = 0.05), but not for soluble tumour necrosis factor receptor 1 (sTNFr1) or neopterin. There were significant correlations between serum C-reactive protein and sIL2ralpha (rs = 0.68, P < 0.0001) and IL-6 (rs = 0.46, P = 0.008) but not sTNFr1 or neopterin levels. Significant differences in appetite, nausea, diet, energy intake, liver function tests and serum levels of metabolic hormones were not detected. CONCLUSION: Weight loss in patients with colorectal liver metastases was not explained by changes in diet, quality of life, or hormones, but activation of the innate and incomplete activation of the acquired immune systems may be involved. Agents that attenuate either the acute-phase inflammatory response or T lymphocyte IL-2 receptor upregulation might reduce weight loss in patients with metastatic disease. PMID- 10361186 TI - Use of a unilateral pudendal thigh flap in the treatment of complex rectovaginal fistula. PMID- 10361187 TI - Comparative study of two sites of colonic conduit placement in the treatment of constipation due to rectal evacuatory disorders. AB - BACKGROUND: Chronic constipation may be treated by antegrade colonic irrigation via a colonic conduit. METHODS: Two alternative sites of colonic conduit construction were evaluated for their effect on the symptoms of 21 consecutive women with intractable constipation primarily due to rectal evacuatory disorders. The conduit was constructed in the sigmoid colon in the first 11 patients and in the transverse colon in the subsequent ten. Symptomatic outcome was evaluated clinically and by questionnaires, with a prospective quality of life assessment in the transverse group. RESULTS: During a median follow-up of 12 (range 6-60) months, reflux or stenosis necessitated revision or dilatation in six patients. Irrigation with a median of 1.3 (0.8-2.0) litres of water achieved evacuation in all patients. Improvements in abdominal pain and bloating were reported by seven of the ten patients in the transverse conduit group, but benefit was found in only three of 11 in the sigmoid group. There was no significant improvement in quality of life scores. In the medium term, seven patients retained a transverse conduit compared with three with a sigmoid conduit. CONCLUSION: The transverse colonic conduit offers better relief from the symptoms of constipation due to rectal evacuatory dysfunction than the sigmoid conduit. PMID- 10361188 TI - Controlled digital anal dilatation under total neuromuscular blockade for chronic anal fissure: a justifiable procedure. AB - BACKGROUND: There is widespread antipathy to digital dilatation of the anus (DDA) for medically resistant anal fissure. A retrospective study was therefore undertaken to test the validity of the criticism of this technique. METHODS: Some 273 patients who underwent DDA for fissure between November 1982 and July 1997 were sent a questionnaire and/or telephoned. Those with impaired control were offered investigation. In addition, routine clinic follow-up data were scrutinized in the 302 available notes of the 307 patients who had undergone DDA for fissure to determine its efficacy. RESULTS: Some 241 patients (88.3 per cent) were contacted successfully a median of 7.8 years after operation. Follow-up records showed the fissure to have healed in 89.1 per cent of 302 patients. No patient was rendered incontinent. Fifteen patients indicated persistently impaired control in the questionnaire, nine (3.8 per cent) as a result of the DDA and six preceding it. All 23 patients who had experienced either temporary or permanent impairment, whether or not pre-existing, were invited to attend for ultrasonography and manometric measurements, of whom 18 accepted. No sphincteric fragmentation was seen, and resting and squeeze pressures did not differ from normal. CONCLUSION: A single DDA appears to heal 89 per cent of chronic anal fissures. Consequent impairment of control is infrequent and minor if the procedure is performed carefully and with the patient paralysed. PMID- 10361189 TI - Comparison of open posterior versus transperitoneal laparoscopic adrenalectomy. AB - BACKGROUND: This study reviewed the results of initial experiences of open posterior adrenalectomy and transperitoneal laparoscopic adrenalectomy in 46 patients. METHODS: Twenty-three adrenalectomies were performed using the open posterior approach. Detailed records of the patients' operative and postoperative progress were compared with those of the first 36 laparoscopic adrenalectomies undertaken for a similar range of conditions. RESULTS: Conversion to laparotomy was necessary in one of 23 open posterior adrenalectomies and five of 36 laparoscopic adrenalectomies. The mean operating time for laparoscopic unilateral adrenalectomy was nearly double that for open surgery (158 versus 85 min). Postoperative complications occurred more frequently in the open adrenalectomy series (12 of 23 versus two of 36) but one late unexplained death followed bilateral laparoscopic adrenalectomy. A mean reduction in hospital stay of 5 days was recorded after laparoscopic adrenalectomy (range 2-5 days for laparoscopic versus 6-11 days for open operation). CONCLUSION: Transperitoneal laparoscopic adrenalectomy was attended by a lower morbidity rate than open adrenalectomy and patients were discharged from hospital more quickly. PMID- 10361190 TI - Epidemiology of Warthin's tumour of the parotid gland in an Asian population. AB - BACKGROUND: Recent studies have documented a number of changing demographic features in the occurrence of Warthin's tumour (adenolymphoma) of the parotid gland. In order to analyse its epidemiology in an Asian population, a retrospective study was performed on all parotid neoplasms (n = 289) operated on between 1988 and 1998. PATIENTS AND METHODS: A total of 209 consecutive patients were selected for study, 136 with pleomorphic adenomas (one bilateral) and 73 with Warthin's tumours (seven bilateral). Patients were analysed with regard to tumour incidence, age, sex and race. Smoking as an aetiological factor in the development of Warthin's tumour was also studied. RESULTS: Warthin's tumour formed 25 per cent of parotid tumours and its ratio to pleomorphic adenomas was 1 : 1.9. Multicentricity was found in 14 patients (19 per cent). The male : female ratio for Warthin's tumours was 4.6 : 1. The proportion of Warthin's tumours did not show any increasing trend relative to pleomorphic adenomas. The racial distribution of Warthin's tumours showed an increased incidence among Chinese and a reduced incidence among Malays and Indians. The adjusted odds ratio for sex and age favouring an association between smoking and Warthin's tumour was 39.5 (95 per cent confidence interval 10.5-149. 0; P < 0.0001). CONCLUSION: The incidence of Warthin's tumour is considerable among Asians although there is still male predominance. There is no rising incidence of Warthin's tumour; the trend parallels the declining smoking rate in the population. The lower incidence among ethnic groups with dark skin seems to suggest concomitant genetic factors other than environmental factors alone in histogenesis. Smokers have a 40-fold greater risk than non-smokers of developing a Warthin's tumour. PMID- 10361191 TI - Case-controlled study of the epidemiological risk factors for breast cancer in Nigeria. AB - BACKGROUND: The incidence of breast cancer is increasing worldwide, more rapidly in societies that hitherto enjoyed a low incidence of the disease, such as most African countries. Most of the epidemiological data on breast cancer from Africa have been retrospective studies with propensity for bias. METHODS: This was a case-controlled study of 250 consecutive patients with breast cancer diagnosed between April 1992 and December 1995. An age- and sex-matched control group of patients with non-oncological and non-endocrine diseases was compared. RESULTS: The peak age incidence of breast cancer in the sample studied was 43 years. There was a statistically significant difference in the height and weight of the patients compared with the controls. Patients also tended to be older at first pregnancy and at first lactation, and had a higher mean number of pregnancies. The patients also tended to be of an early birth order, to have lactated less often, to have used contraceptives and to have abused alcohol compared with the controls. CONCLUSION: The incidence of breast cancer in this environment is increasing. This is partly a result of the changing demographic profile, acquisition of 'western' lifestyle, and the changing socioeconomic profile of the country. PMID- 10361192 TI - Unique distribution patterns of metastatic lymph nodes in patients with superficial carcinoma of the thoracic oesophagus. AB - BACKGROUND: Lymph node metastasis is commonly found in carcinoma of the thoracic oesophagus, even when the tumour invades only the submucosa. Although lymph node status greatly influences the outcome, the pattern of early lymphatic spread has not been investigated, and the role of lymph node dissection is still a matter of controversy. METHODS: A series of 110 patients with superficial carcinoma who underwent systematic extended lymph node dissection was investigated retrospectively. RESULTS: Lymph node involvement was found in 0 per cent (none of nine), 23 per cent (five of 22) and 49 per cent (38 of 78) of tumours that invaded the lamina propria, muscularis mucosa and submucosa respectively. Anatomically distant lymph nodes (recurrent nerve nodes and perigastric nodes) were involved more frequently than other intrathoracic nodes adjacent to the main tumour. Only three patients had involvement limited to the intrathoracic group, and in carcinoma that invaded only the muscularis mucosae, all metastatic nodes were located at the thoracocervical junction or in the abdomen. The 5-year survival rate was 89 per cent in the node-negative group and 54 per cent in the node-positive group (P < 0.0003). CONCLUSION: The recurrent nerve nodes and perigastric nodes are the principal proximal regional lymph nodes involved in superficial carcinoma of the thoracic oesophagus. Systematic lymph node dissection, which included these nodes, yielded an acceptable and favourable outcome in patients with node-positive superficial carcinoma. PMID- 10361193 TI - Clinical significance of telomerase activity in the non-cancerous epithelial region of oesophageal squamous cell carcinoma. AB - BACKGROUND: The aim of this study was to examine telomerase activity in affected and adjacent tissue in patients with oesophageal squamous cell carcinoma (SCC). METHODS: Telomerase activity was measured in oesophageal SCC cells, normal oesophageal culture cells, primary cancer tissues and adjacent normal tissues from patients with oesophageal SCC using a polymerase chain reaction-based assay. RESULTS: All oesophageal SCC cells had telomerase activity, whereas normal cultured cells showed no activity. All 57 cancer tissues showed telomerase activity compared with only five (10 per cent) of 50 normal tissues. Cancer cells infiltrating the vessels of mucosal or submucosal tissues in non-cancerous regions were detected in four of five telomerase-positive normal tissues, whereas such infiltration was detected in only three of 45 telomerase-negative normal tissues. CONCLUSION: In patients with oesophageal SCC, measurement of telomerase activity in normal epithelium is a highly sensitive method of detecting the microinvasion of cancer cells. PMID- 10361194 TI - Impaired healing of extraperitoneal intestinal anastomoses. AB - BACKGROUND: The extra-anatomical position of a cervical oesophagogastrostomy might be a reason for impaired anastomotic healing. METHODS: This hypothesis was tested in a rat model. Jejunal resection with an end-to-end jejunojejunostomy was placed intra-abdominally in group 1 (n = 24) and subcutaneously in group 2 (n = 30). Jejunum without anastomosis was placed subcutaneously in group 3 (n = 12). After 3, 7 or 14 days the rats were killed; the bursting pressure of the anastomosis or jejunum was measured and the hydroxyproline level was determined. RESULTS: Two of 24 rats in group 1 and eight of 30 in group 2 died following anastomotic leakage (P not significant) and were excluded from other measurements. Bursting pressure was decreased after 3 days in group 1 (mean(s.e.) 62(10) mmHg) and group 2 (57(10) mmHg) compared with that in group 3 (204(17) mmHg) (P < 0.001). After 7 days, it was in the normal range in group 1 (200(14) mmHg), but lower in group 2 (104(15) mmHg) compared with that in group 3 (230(8) mmHg) (P < 0.001). Differences in hydroxyproline levels were not statistically significant between the groups after 3, 7 and 14 days. CONCLUSION: Healing of jejunojejunostomies is impaired in an extraperitoneal position compared with an intra-abdominal position. PMID- 10361195 TI - Clinical evaluation of lymph node metastasis in gastric cancer defined by the fifth edition of the TNM classification in comparison with the Japanese system. AB - BACKGROUND: This study compared the classification of lymph node metastasis according to the number of involved nodes based on the new tumour node metastasis (TNM) system (fifth edition) with the classification by the Japanese Research Society for Gastric Cancer from an anatomical perspective. METHODS: The two classifications were related to long-term results in 1489 patients with gastric cancer who underwent gastrectomy with systematic extended lymphadenectomy. RESULTS: Both classifications performed well as prognostic indicators (5-year survival rates: pathological (p) N0, 89 per cent; pN1, 66 per cent; pN2, 34 per cent; pN3, nil; and M1, 10 per cent by the TNM classification; n0, 89 per cent; n1, 63 per cent; n2, 46 per cent; n3, 20 per cent; and n4, 8 per cent by the Japanese classification). For regional lymph nodes, the TNM classification was a better index of the prognosis. Significant survival differences were observed among patients with M1 disease according to the number of involved lymph nodes (between one and six nodes, 48 per cent; seven to 15 nodes, 12 per cent; more than 15 nodes, 2 per cent), indicating that patients with distant metastatic lymph nodes (M1) should also be classified by the number of involved nodes. On the other hand, the Japanese classification has the added benefit of being a good indicator of the anatomical extent of lymphadenectomy. CONCLUSION: The new TNM classification provided a better index of the prognosis of patients who underwent systematic lymph node dissection. However, both classifications have specific benefits in the surgical treatment of gastric cancer. PMID- 10361196 TI - Vascular surgical society of great britain and ireland: twenty-three years of experience of carotid endarterectomy AB - BACKGROUND: Carotid endarterectomy (CEA) has been an evidence-based treatment for symptomatic severe carotid stenosis since 1991. Surgical techniques and patient selection have changed over the years. The results of CEA in a single centre over a 23-year period were reviewed. METHODS: Prospectively gathered preoperative, operative, postoperative and long-term follow-up data were analysed. Routine intraoperative shunting and patch closure has been used since 1988. Data were analysed using the chi2 test or by logistic regression, adjusting for age at operation and date of operation. RESULTS: Five hundred and seventy-three CEAs (37 bilateral and three repeat procedures) were carried out on 533 patients. Trainees performed an increasing proportion of CEAs from 1996 to 1998 (15, 50 and 56 per cent respectively). The perioperative death rate was 0.8 per cent and the rate of any perioperative neurological deficit was 6.9 per cent. Other causes of morbidity included nerve injury (5.1 per cent) of which the commonest was to the hypoglossal nerve (2.7 per cent). During follow-up (median 4 (range 0-22) years) there were 81 neurological events (15.9 per cent) which included 35 ipsilateral (6.6 per cent) and 18 contralateral (3.4 per cent) strokes. There was no significant difference in outcome for grade of surgeon, intraoperative shunting or patch closure. Major causes of death were cardiac death (74; 14.6 per cent) followed by stroke (23; 4.5 per cent) and cancer (20; 3.9 per cent). CONCLUSION: The introduction of routine intraoperative shunting and patch closure, as well as allowing surgical trainees to perform supervised CEAs, has not affected perioperative morbidity and mortality rates or long-term outcome. PMID- 10361197 TI - Vascular surgical society of great britain and ireland: thrombelastography can differentiate ischaemic from haemorrhagic stroke AB - BACKGROUND: Thrombolysis could be a major step forward in the treatment of acute ischaemic stroke. Early treatment is essential to maximize therapeutic benefit. Imaging by computed tomography (CT) or magnetic resonance imaging is mandatory to exclude haemorrhagic stroke before initiation of therapy. Thrombelastography (TEG), a test of global haemostasis, produces a characteristic tracing over 15-30 min. The potential of TEG to differentiate patients with ischaemic from haemorrhagic stroke was investigated. METHODS: Fifteen patients with a clinical diagnosis of acute stroke were studied. Fibrinogen levels and lipid profiles were measured, and CT of the brain, coagulation screens and TEG were performed. The CT scans were interpreted by a single radiologist. TEG data were classified as normal (index less than 2), hypercoagulable (index 2-3) or profoundly hypercoagulable (index greater than 3). RESULTS: There was no correlation between any of the parameters studied other than TEG with CT findings. Both patients with a haemorrhagic stroke showed normal findings on TEG. Eleven of the 12 patients with ischaemic stroke were hypercoagulable or profoundly hypercoagulable. CONCLUSION: TEG is capable of differentiating haemorrhagic from ischaemic stroke. It has the potential to target thrombolytic therapy for those patients most likely to benefit. PMID- 10361198 TI - Vascular surgical society of great britain and ireland: transcranial doppler directed dextran therapy in the prevention of postoperative carotid thrombosis AB - BACKGROUND: Evidence suggests that embolization precedes carotid thrombosis, a previously unpredictable event complicating 2-3 per cent of all carotid endarterectomies. It was hypothesized that dextran 40 therapy might prevent progression to complete thrombosis in high-risk patients. METHODS: Between October 1995 and July 1998, 400 consecutive patients were monitored following carotid endarterectomy using transcranial Doppler ultrasonography. Those with sustained embolization (more than 25 in 10 min) or those with emboli that distorted the middle cerebral artery waveform were commenced on an incremental dextran 40 infusion. RESULTS: Two hundred and sixteen patients (54 per cent) had one or more emboli detected (96 per cent within 2 h of flow restoration) but only 15 (4 per cent) required dextran therapy. Embolization ceased in each case although the dextran dose had to be adjusted in four. In one of the latter patients, embolization recurred on day 5 but was again controlled with high-dose dextran. Overall, the death and any stroke rate was 2 per cent and no patient suffered a stroke due to carotid thrombosis. CONCLUSION: A few patients develop sustained embolization following carotid endarterectomy which, in previous studies, has been shown to be highly predictive of carotid thrombosis. The authors' experience to date suggests that dextran can stop this phase of embolization and prevent progression to complete carotid thrombosis. However, the dose of dextran has to be adjusted in 25 per cent of patients (i.e. blind administration of dextran may not be effective) and, very rarely, embolization may recur later. PMID- 10361199 TI - Vascular surgical society of great britain and ireland: deaths from ruptured abdominal aortic aneurysm in wales AB - BACKGROUND: The aim was to determine the true incidence and operative mortality rate of patients with ruptured abdominal aortic aneurysm (AAA) who reach hospital alive in Wales. METHODS: Patients presenting with a ruptured AAA between September 1996 and August 1997 were analysed. The data were collected prospectively by an independent body, observing strict confidentiality. RESULTS: Two hundred and thirty-three patients with confirmed ruptured AAA were identified. One hundred and thirty-three patients (57 per cent) underwent attempted operative repair. Eighty-five (64 per cent) died within 30 days. All 100 patients who received no operation died. Of the 233 patients, 92 were admitted under vascular surgeons (VSs) and 141 under non-vascular surgeons (NVSs). VSs operated on 82 patients (89 per cent) of whom 50 (61 per cent) died; NVSs operated on 51 (36 per cent) of whom 35 (69 per cent) died. CONCLUSION: This study is the only independent prospective study of death among patients with ruptured AAA who reached hospital alive. Some 57 per cent of the patients with a ruptured AAA were operated on. The operative mortality rate was 64 per cent and the overall mortality rate was 79 per cent. VSs were significantly more aggressive (89 per cent) in the management of ruptured AAA (i.e. more likely to operate) than NVSs (36 per cent) (P < 0.0001). Despite this, the operative mortality rate for VSs was 61 per cent, whereas for NVSs it was 69 per cent (P = 0.372). The overall mortality rate (including operated and non-operated patients) for NVSs (89 per cent) was significantly higher than that for VSs (65 per cent) (P < 0.0001). In Conclusion:, ruptured AAA is common in Wales and associated with a high mortality rate even when managed by VSs. PMID- 10361200 TI - Vascular surgical society of great britain and ireland: distal venous arterialization for non-reconstructable arterial disease AB - BACKGROUND: Despite advances in vascular surgical and interventional radiological techniques, 14-20 per cent of patients with critical lower limb ischaemia will not be suitable for distal arterial reconstruction owing to occlusion of crural and pedal vessels. Following investigation into the venous anatomy of the foot, the disease-free venous bed was used as an alternative conduit for perfusion of peripheral tissues. METHODS: For 22 months, venous arterialization was performed for all patients with non-reconstructable disease but potentially salvageable feet. There were 15 patients; 14 had surgery for chronic ischaemia and one for acute embolization from a popliteal aneurysm after unsuccessful thrombolysis. The inflow vessel was the common femoral artery in 12 and the popliteal artery in three patients. Reversed contralateral long saphenous vein was used in five, a composite graft in three and a polytetrafluoroethylene graft with vein patch in the remaining seven. The distal anastomosis was to the dorsal venous arch (13 cases) and in two to vena comitantes of the posterior tibial vein owing to extensive tissue loss. Various techniques were employed to destroy the valve. RESULTS: Three procedures failed immediately after operation and major amputation was required. The remaining 12 patent grafts were monitored by duplex scan; two required intervention to correct distal anastomotic stenosis by angioplasty and atherectomy (one patient each). The limb salvage rate at 2-22 months of follow-up was 80 per cent. CONCLUSION: Distal venous arterialization is a unique and promising operation which merits further evaluation. PMID- 10361201 TI - Vascular surgical society of great britain and ireland: In situ replacement of infected aortic grafts with rifampicin-bonded prostheses AB - BACKGROUND: Prosthetic graft infection following abdominal aortic aneurysm (AAA) surgery is life threatening. Treatment options include total graft excision and extra-anatomic bypass or in situ replacement of the graft. The latter option is gaining popularity but the long-term outcome remains uncertain, particularly in light of the increasing prevalence of methicillin-resistant Staphylococcus aureus (MRSA) infection. A retrospective study was undertaken to assess the outcome after graft excision and in situ replacement with a rifampicin-bonded prosthesis for the treatment of major aortic graft infection. The prevalence of MRSA and its influence on outcome was also assessed. METHODS: Over 6 years between January 1992 and December 1997, 11 patients (eight men) with major aortic graft infection underwent total graft excision and in situ replacement with a rifampicin-bonded prosthesis. RESULTS: The median age was 66 (range 49-78) years. Four patients presented with haemorrhage from an aortoenteric fistula, three had retroperitoneal abscess, two had graft occlusion, one had a perigraft collection on computed tomography and one presented with a ruptured suprarenal false aneurysm. Organisms were cultured from ten patients. Staphylococcus epidermidis and Streptococcus faecalis were predominant. MRSA was isolated from two patients, both of whom had originally undergone ruptured AAA repair. Two patients died and three suffered non-fatal complications. Two patients died during follow-up, one from suspected ongoing MRSA infection and the other from recurrent graft infection. Seven patients remain alive and clinically free from infection, although two were lost to follow-up at 12 and 15 months. CONCLUSION: The long term results following total graft excision and in situ replacement with a rifampicin-bonded prosthesis appear to be favourable. MRSA infection seems to be associated with a poor prognosis. PMID- 10361202 TI - Vascular surgical society of great britain and ireland: total abdominal approach for repair of type IV thoracoabdominal aortic aneurysm AB - BACKGROUND: The total transabdominal approach for thoracoabdominal aneurysm (TAA) was described in 1995 and it was suggested that outcome might be improved if the chest was not opened. This study reports early results of the technique with respect to operative morbidity and mortality rates for patients with abdominal aortic aneurysm extending to the diaphragm. METHODS: Between 1995 and 1998, 26 patients (median age 71 (range 52-84) years) underwent repair of a type IV TAA using a total abdominal approach and medial visceral rotation. RESULTS: Three patients presented with a contained leak. All survived but one developed paraplegia. Other complications included chest infection (five patients), myocardial infarction (three), reoperation for bleeding (three) and temporary dialysis in one patient. There were three perioperative deaths, two from myocardial infarction and one from multisystem organ failure. CONCLUSION: The total abdominal approach for the repair of type IV TAA is a reasonable alternative to a full thoracoabdominal incision. Thoracic complications are minimized, renal and visceral ischaemia times are low, and the perioperative mortality rate is acceptable. PMID- 10361203 TI - Vascular surgical society of great britain and ireland: changes in proximal aortic neck dimensions following endovascular repair of abdominal aortic aneurysm AB - BACKGROUND: Dilatation of the proximal neck following conventional open repair of abdominal aortic aneurysm (AAA) has been reported. Such continued dilatation following endovascular repair (EVR) could potentially be a disaster resulting in graft slippage, endoleak and aneurysm rupture. The aim of this study was to detect any change in proximal neck diameter following EVR of AAAs. METHODS: One hundred patients had undergone EVR of an AAA over a 4-year period in whom contrast-enhanced spiral computed tomography was performed both before and after EVR (1 week, 3, 6 and 12 months and annually thereafter). Change in aortic proximal neck diameter, change in maximum aortic diameter, presence of endoleaks, and change in length from the lowest renal artery to the aortic bifurcation was sought. RESULTS: The median anteroposterior and transverse diameter decreased from 64 mm before operation to 56 and 54 mm respectively after operation. This trend in reduction in maximum diameter was not seen in patients with endoleaks. There was no significant change in proximal neck diameter when measured at 5-mm intervals following EVR. There was also no significant change in the aortic length following EVR. CONCLUSION: There was no evidence of proximal neck dilatation or aneurysm length reduction following EVR of AAAs. PMID- 10361204 TI - Vascular surgical society of great britain and ireland: immunoglobulin A antibodies against chlamydia pneumoniae are associated with expansion of small abdominal aortic aneurysms and declining ankle blood pressure AB - BACKGROUND: The potential correlation between chronic infection with Chlamydia pneumoniae and the progression of small abdominal aortic aneurysms (AAAs) and lower limb atherosclerosis was studied. METHODS: Mass screening for AAA was carried out in outdoor clinics at all hospitals in the county. Some 139 men (aged 65-73 years) with a 3.0-4.9-cm AAA were followed prospectively for 1-3 (mean 2.7) years. Initially, an interview and examination was performed, and blood samples were taken. RESULTS: Some 62 per cent (53-71 per cent) had an immunoglobulin (Ig) A level of 40 or more, or an IgG level of 64 or above. Some 83 per cent (74-93 per cent) had an IgA level of 20 or more, or an IgG level of 32 or more. Men with an IgA level of 20 or more had 51 per cent greater AAA expansion and men with an IgA level of 40 or above had 24 per cent more expansion. An IgA level of 20 or more, or IgA of 40 or greater, were significant independent predictors of AAA expansion adjusted for age, smoking, initial AAA size, steroid treatment, diastolic blood pressure, pulmonary function and other plasma factors. The ankle blood pressure index (ABI) of the IgA-seropositive men decreased 11 per cent, while the ABI decreased by 5 per cent among IgA-seronegative men (P < 0.05). The significant difference persisted after adjusting for age, smoking, initial systolic ankle blood pressure, initial brachial systolic or diastolic blood pressure, but disappeared after adjusting for low-density lipoprotein (LDL) levels. CONCLUSION: A high proportion of men with a small AAA have signs of chronic C. pneumoniae infection. The progression of AAAs and lower limb atherosclerosis seems to be correlated to chronic infection with C. pneumoniae. PMID- 10361205 TI - Vascular surgical society of great britain and ireland: management of coexisting intra-abdominal disease in aortic surgery AB - BACKGROUND: The management of coexisting intra-abdominal disease in aortic surgery is controversial. A staged repair is preferred by many and, in general, the symptomatic lesion is treated first. Twenty-one years ago this vascular unit elected a policy of treating such lesions synchronously. This was on the theoretical premises that the benefits of a single operation in these patients would outweigh the potential risks of graft infection and increased complication rates from a prolonged procedure. METHODS: The case records of 676 patients undergoing aortic grafting for aneurysmal or occlusive disease between 1978 and 1998 were analysed retrospectively. RESULTS: Fifty-five patients (8 per cent) had coexisting intra-abdominal diseases treated at the time of aortic graft surgery. These included biliary disease (26), and gastric (12), intestinal (13), urological (two), hepatic (one) and splenic (one) pathology. The median age at presentation was 72 (range 46-90) years. There were three hospital deaths and five patients required early reoperation, three for lower limb ischaemia and two for intra-abdominal bleeding. One patient developed a subphrenic abscess and there were three superficial wound infections. There were no graft infections in this group of 55 patients. CONCLUSION: This large single-centre experience with synchronous intra-abdominal surgery and aortic grafting demonstrates that it is safe and does not predispose to an increased risk of graft infection or perioperative haemorrhage. PMID- 10361206 TI - Vascular surgical society of great britain and ireland: outcome after ligation of infected false femoral aneurysms in intravenous drug abusers AB - BACKGROUND: Infected false femoral aneurysm (IFFA) is a life-threatening complication of intravenous drug abuse and presents a difficult management problem for the vascular surgeon. Controversy exists regarding the best management. The choice lies between ligation and excision with immediate revascularization, and ligation and excision with observation; reconstruction is reserved for critical ischaemia. METHODS: After disappointing results with the former method it was decided to perform ligation and excision with observation as the initial treatment of IFFA. A 9-year experience of 28 patients treated at this hospital is reviewed. RESULTS: In 26 cases of primary ligation and excision of an IFFA there were no amputations and patients described claudication only in follow up. In two cases of a second IFFA in the same limb, repeat ligation and excision resulted in one viable limb with claudication only and one above-knee amputation for non-viability. At 9-year follow-up (80 per cent complete), over 90 per cent of the patients were still drug abusers and therefore not suitable for revascularization. There were two deaths, both of which were drug related. CONCLUSION: Ligation and excision of an IFFA is simple, effective and safe, and is the most appropriate method of dealing with these challenging patients. PMID- 10361207 TI - Vascular surgical society of great britain and ireland: mid-term results of arterial allograft below-knee bypasses for limb salvage AB - BACKGROUND: Arterial allografts (AAs) have been recently reconsidered in the treatment of critical limb ischaemia when vein material is absent, because of the disappointing results with artificial grafts. The aim of this study was to report the results observed in three centres where AAs were used for infrainguinal reconstruction in critical limb ischaemia. METHODS: Between 1991 and 1997, 165 AA bypasses were performed in 148 patients (90 men) with a mean age of 70 (range 20 93) years. Indications for operation were rest pain in 54 patients and tissue loss in 111. Mean resting ankle pressure was 53 mmHg in 96 non-diabetic patients and mean transcutaneous partial pressure of oxygen was 10 mmHg in 52 diabetic patients. AAs were obtained from cadaveric donors. The distal anastomosis was to the below-knee popliteal artery in 34 cases, to a tibial artery in 114 and to a pedal artery in 17. RESULTS: At 30 days, the mortality rate was 3 per cent, primary patency 83 per cent, secondary patency 90 per cent and limb salvage rate 98 per cent. During follow-up (mean 31 months) 65 grafts failed primarily. Causes of primary failure were thought to be progression of the distal disease in 15 cases, myointimal hyperplasia in 16, graft degradation in ten (four dilatations, three stenoses, two ruptures and one dissection), other causes in eight and not known in 16. Primary and secondary patency rates at 3 years were 35(s.e. 9) per cent and 42(s. e. 10) per cent. The limb salvage rate at 3 years was 76(s.e. 8) per cent. CONCLUSION: AAs lead to a good foot salvage rate but poor patency rates. The results are similar to those obtained with polytetrafluoroethylene grafts. PMID- 10361208 TI - Vascular surgical society of great britain and ireland: contribution of malnutrition to postoperative morbidity in vascular surgical patients AB - BACKGROUND: In an elderly population of surgical patients, poor mobility, poor diet and chronic disease contribute to a significant risk of malnutrition. Malnutrition is associated with muscle weakness, fatigue, poor wound healing and immunological dysfunction. The aim of the study was to establish the prevalence of malnutrition in vascular surgical patients and to compare postoperative infection rates in well nourished and malnourished patients. METHODS: A nutritional assessment was performed on 71 patients (49 men; median age 65 (range 26-85) years) attending preassessment for vascular surgical procedures. Nutritional status was measured using validated indicators of malnutrition: estimated weight changes over 3 months; body mass index; mid-arm muscle circumference (MAMC) calculated using triceps skin fold thickness (TSF) and mid arm circumference (MAC) (MAMC = MAC - (3.14 x TSF)); and serum albumin concentration. Fifty-nine patients were followed after vascular surgery. The incidence of postoperative infections was related to preoperative nutritional status. RESULTS: Nineteen patients (27 per cent) had normal values for all nutritional indicators examined. The remaining 52 patients (73 per cent) had one (37), two (12), three (two) or four (one) nutritional indicators within the range for malnutrition. Among the 59 patients who underwent surgery there were five chest infections, seven wound infections, one urinary tract infection and one infected central line in 13 patients following six femorodistal bypasses, four abdominal aortic aneurysm repairs and three miscellaneous arterial procedures. The incidence of septic complications was zero in 14 patients with normal nutritional indicators and 41 per cent (13 of 32) in patients with indicators of malnutrition (P < 0.05, Fisher's exact test). CONCLUSION: Malnutrition is prevalent among vascular patients and may contribute to postoperative morbidity. Malnourished patients should be identified and referred to the dietician at the earliest opportunity to minimize the morbid effects of undernutrition. PMID- 10361209 TI - Vascular surgical society of great britain and ireland: integrated care pathways for vascular surgery AB - BACKGROUND: Integrated care pathways (ICPs) represent a multidisciplinary approach to clinical patient care. METHODS: A 1-year prospective trial of the use of ICPs for elective vascular surgical procedures was undertaken. A multidisciplinary group constructed ICPs for patients admitted for open repair of abdominal aortic aneurysm, carotid endarterectomy or femoropopliteal bypass grafting. Patient management followed ICPs on a daily basis with signatures required to confirm that each action had been taken. Variances from the ICPs were carefully recorded. Audit of variance data allowed subsequent revision of the ICPs and hence provision of the best possible nursing and clinical practice. METHODS: A total of 33 patients were entered into the study; 16 had a femoropopliteal bypass graft, eight carotid endarterectomy and nine open repair of an abdominal aortic aneurysm. ICPs were well received by patients and staff. They improved communication, promoted an appreciation of each health group's role in patient care, increased nursing autonomy, reduced calls to junior medical staff, improved patient education and confidence, and caused a marked reduction in length of hospital stay. Overall, patients were discharged 13 per cent earlier after open abdominal aortic aneurysm repair, 22 per cent earlier after carotid endarterectomy and 38 per cent earlier after femoropopliteal bypass grafting. CONCLUSION: ICPs have clear benefits. They improve overall clinical efficiency and enhance clinical governance. Successful use of ICPs depends upon 'clinical champions' and effective project management. Sufficient resources and training are essential. PMID- 10361210 TI - Vascular surgical society of great britain and ireland: systematic review of intra-arterial thrombolytic therapy for peripheral vascular occlusions AB - BACKGROUND: A systematic review of intra-arterial thrombolytic therapy for peripheral vascular occlusions was undertaken. METHODS: The four major electronic databases (Medline, Cinhal, Embase and The Cochrane Library) were searched to identify randomized controlled trials of thrombolytic therapy in the treatment of limb ischaemia. The search was limited to English language articles, or those that provided a sufficiently detailed English summary, and to articles published after 1980. Trials were considered for inclusion if they were randomized controlled trials, systematic reviews or meta-analyses. Data were extracted independently by two reviewers and aggregate outcomes were obtained using a random effects meta-analysis. RESULTS: A total of 32 articles were found but only 12 were assessed to be of sufficient quality to be included in the review. There was a distinct lack of large randomized controlled trials comparing thrombolysis with surgical management. The exceptions were the Thrombolysis or Peripheral Arterial Surgery (TOPAS) and Surgery versus Thrombolysis for Ischemia of the Lower Extremity (STILE) trials, but both had some methodological flaws. The STILE trial did not achieve the sample size originally determined and the TOPAS trials had a large number of contributing centres (113) compared with the sample size (548). Meta-analysis showed no significant differences between thrombolysis and surgery in terms of major amputation (relative risk (RR) 0.9 (95 per cent confidence interval 0.6-1.4)) and death (RR 1.2 (0.8-1.9)). However, there was an increased risk of residual ischaemia with thrombolysis (RR 1.8 (1.2-2.5)) and haemorrhage (RR 2.9 (1.1-7.9)). Patients who were shown to benefit from thrombolysis were those with short-duration ischaemia (RR reduction, 72 per cent; numbers needed to treat, three) and occluded grafts (RR reduction, 58 per cent; numbers needed to treat, four). CONCLUSION: Despite the theoretical advantages of thrombolysis, there is still insufficient evidence to justify its widespread use except in patients with graft occlusions and short-duration ischaemia. PMID- 10361211 TI - Vascular surgical society of great britain and ireland: duplex surveillance does not enhance infrainguinal prosthetic bypass graft patency AB - BACKGROUND: All patients discharged from this unit with a patent infrainguinal polytetrafluoroethylene (PTFE) bypass are entered into a graft surveillance programme. Previous internal audit had suggested that duplex surveillance was not worthwhile in this patient group. The aim of this study was to compare the fate of infrainguinal PTFE grafts before and after stopping duplex surveillance. METHODS: Between January 1986 and December 1994, 220 grafts (141 above-knee popliteal, 69 below-knee popliteal, ten tibial) were entered into the duplex surveillance programme. Between January 1995 and June 1996, 56 further grafts (29 above-knee popliteal, 21 below-knee popliteal, six tibial) were followed prospectively without duplex scans. RESULTS: During the first study interval, an 'abnormal' scan was reported in 66 of 220 grafts. For clinical reasons (no further reconstruction feasible), no intervention was undertaken in 56 patients. Of these, 34 grafts occluded and 17 amputations were performed. An intervention to maintain patency was performed in ten patients. In 154 patients with 'normal' scans, 53 grafts occluded and 21 amputations were performed. During the second study interval, 20 grafts occluded and 12 amputations were performed. In six patients, an attempt was made to re-establish patency and this was successful in two. Kaplan-Meier 36-month primary and secondary patency rates were 48 and 51 per cent respectively for the group that underwent duplex surveillance, and 58 and 60 per cent for the group that was followed without duplex imaging. CONCLUSION: Prospective duplex surveillance of infrainguinal PTFE bypass grafts does not enhance graft patency and cannot be justified. PMID- 10361212 TI - Vascular surgical society of great britain and ireland: isolated femoral profundoplasty with endarterectomized superficial femoral artery for limb salvage in the elderly AB - BACKGROUND: This longitudinal observational study was designed to show that isolated profundoplasty to relieve a significant stenosis is a valid procedure for limb salvage in the elderly. METHODS: Twenty-seven patients with critical limb ischaemia underwent isolated profundoplasty using endarterectomized superficial femoral artery as an arterial patch. Nineteen patients were men. The mean age was 72 (range 65-79) years. All patients had rest pain with or without an ischaemic foot ulcer or pedal gangrene. RESULTS: There was no operative death or immediate operative failure. All 27 limbs were improved, with relief of rest pain, healing of ulcers and healing after minor foot amputations. The 27 patients underwent a mean of 30 (range 12-45) months of periodic observation in an outpatient clinic, either to the present time or until death. Two patients required late amputations and one a femorodistal bypass. CONCLUSION: Isolated profundoplasty has a place in leg revascularization in the high-risk elderly patient. Endarterectomized superficial femoral artery as a patch has the advantages of almost universal availability and anatomical convenience. PMID- 10361213 TI - Vascular surgical society of great britain and ireland: two-year follow-up after acute thromboembolic lower limb ischaemia: the importance of continuing warfarin treatment AB - BACKGROUND: The Vascular Surgical Society carried out an audit of follow-up after acute thromboembolic lower limb ischaemia. METHODS: The audit of acute lower limb ischaemia was done between 1 January and 31 March 1996 and included 474 patients. This report describes a 2-year follow-up of patients with a diagnosis of thrombosis or embolism who survived 30 days. Details were obtained for 214 (75 per cent) of 287 patients: half (107) were men, and the age range was 21-96 (mean 74) years. RESULTS: Amputation of the leg affected initially had been required (after 30 days) in 12 per cent. Further acute leg ischaemia had occurred in 11 per cent, and 9 per cent had undergone arterial reconstruction (11), angioplasty (seven) or thrombolysis (two). Major medical events were reported in 31 per cent and 35 per cent had died during the 2-year period. After the episode of acute ischaemia, warfarin was started in 57 per cent of patients, but was continued in only 43 per cent. Reasons for stopping were seldom known, nor was duration of treatment (range 1-22 (median 6) months in reported cases). Significantly more patients who were not on warfarin suffered recurrent acute limb ischaemia (19 versus 3 per cent; P < 0.01). No significant association was found between recurrent acute ischaemia and initial diagnosis of embolism, thrombosis or atrial fibrillation (but numbers of recurrences in each group were small). CONCLUSION: The incidence of recurrent acute thromboembolic limb ischaemia up to 2 years was significantly lower in patients treated with warfarin. Warfarin treatment is often stopped for no clear reason, especially in the elderly. Explicit advice about long-term warfarin therapy should be given when these patients are discharged from hospital. PMID- 10361214 TI - Vascular surgical society of great britain and ireland: patients with a failed infrainguinal bypass graft have abnormal lipid metabolism AB - BACKGROUND: Dyslipidaemias adversely affect vascular tone, endothelial function and platelet activation. Abnormal lipid metabolism has not been established as a risk factor for infrainguinal bypass graft failure. Lipid metabolism was evaluated prospectively in patients with patent and occluded grafts. METHODS: Twenty-eight patients with failed infrainguinal grafts (group 1) were identified from a prospective computerized database. Twenty matched controls with functioning grafts (group 2) were recruited from a graft surveillance programme. Fasting blood samples were analysed for triglyceride (TG), high-density lipoprotein (HDL) and cholesterol. A newly devised rapid TG tolerance test was conducted with analysis of TG at 1, 2 and 2.5 h. Endothelial lipoprotein lipase (LPL) was measured 30 min after intravenous administration of heparin 50 units kg 1. RESULTS: The cholesterol : HDL ratio was significantly higher in group 1 than group 2 (median 6.0 (range 3. 1-10.0) versus 4.7 (2.3-9.4); P = 0.0008, Mann Whitney test) as was fasting TG (2.6 (1.0-6.3) versus 1.7 (0.6-4.7) mmol l-1; P = 0.03). The area under the curve of the TG tolerance test was not significantly different (P = 0.08); however, the 2.5-h levels of TG were significantly different between the groups (group 1, 2.0 (0.5-6. 6) mmol l-1; group 2, 1.2 (0.3 6.8) mmol l-1; P = 0.01). LPL was significantly lower in group 1 (52.4 (2.2 235.9) versus 86.8 (28. 6-281.5) mmol/l; P = 0.04). CONCLUSION: These data suggest that abnormal lipid metabolism is a significant risk factor for infrainguinal graft occlusion. This study provides a rationale for randomized trials of lipid-modifying therapies in patients undergoing arterial reconstruction for peripheral vascular disease. PMID- 10361215 TI - Vascular surgical society of great britain and ireland: platelet function during carotid endarterectomy and the antiplatelet effect of dextran 40 AB - BACKGROUND: Dextran 40 has been shown to reduce cerebral embolization following carotid endarterectomy (CEA). This study aimed to determine changes in platelet function during CEA and the antiplatelet effect of dextran 40. METHODS: Platelet function was measured by whole-blood flow cytometry in the peripheral arterial blood of patients during CEA and 24 h later. The binding index of P-selectin and PAC-1 expression were measured as markers of activation and aggregation. Patients were kept on aspirin 75-150 mg until the day of surgery and received an intravenous bolus of 5000 units unfractionated heparin before carotid artery clamping. High-intensity transient signals (HITS) in the ipsilateral middle cerebral artery were measured with transcranial Doppler (TCD) ultrasonography before, during and after operation. Results are presented as median (interquartile range) and statistical significance was determined using the Mann Whitney U test. RESULTS: Thirty-eight patients undergoing CEA were studied. The P selectin binding index rose significantly from incision (0.9 (0.2-2.7)) after carotid clamping (1.5 (0.6-3.6); P < 0.01), clamp release (1.7 (0. 3-3.0); P < 0.01), 1 h after operation (1.5 (0.3-2.6); P < 0.05) and 24 h after operation (1.3 (0.6-2.5); P < 0.05). PAC-1 binding index increased from incision (0.4 (0.1 0.8)) after carotid clamping (2.0 (0.4-4.2); P < 0.01) and clamp release (1.8 (0.3-2.9); P < 0.05). TCD monitoring showed an increase in preoperative HITS per 30 min (2 (0-3)) during dissection (8 (1-15); P < 0.05), after clamp release (16 (2-27); P < 0.01) and during recovery (10 (2-29); P < 0.01). After operation, patients with more than 50 HITS per 30 min were started on an infusion of dextran 40. Six patients had a dextran 40 infusion. The P-selectin binding index decreased from 1.6 (0.7-1.9) to 0.6 (0.3-1.5) 1 h after dextran (P < 0.05) and 0.1 (0.1-0.2) 24 h after dextran (P < 0.05). PAC-1 expression decreased from 0.4 (0.3-0. 5) to 0.1 (0.1-0.1) 24 h after dextran (P < 0.05). CONCLUSION: Significant platelet activation and aggregation occurs during CEA despite the use of antiplatelet treatment. A simultaneous increase in HITS was demonstrated with TCD. Dextran 40 was shown to have an antiplatelet effect after CEA; this is further evidence that it may have a role in reducing thromboembolic complications. PMID- 10361216 TI - Vascular surgical society of great britain and ireland: scottish audit of surgical mortality, 1997 AB - BACKGROUND: A national audit of surgical deaths can be seen as the final step in what has been termed the 'journey of care' for both the individual patient and for the population as a whole. METHODS: The Scottish Audit of Surgical Mortality (SASM) examines all hospital deaths in Scotland occurring while under the care of a surgical specialist and all deaths within 30 days of an operation. RESULTS: Compliance for the completion of forms relating to vascular deaths during the first 4 years of the SASM (1994-1997) has remained over 92 per cent. In 1997 the compliance was 97 per cent. These figures compare favourably with the other surgical specialties where the overall compliance for 1997 was 92 per cent. Some 14 per cent of the 4408 deaths reported in 1997 occurred in patients admitted with vascular disease, the majority (83 per cent) being emergency admissions. Of these 507 vascular deaths, 293 were postoperative; the remaining 214 patients did not undergo operation. This represents an improvement over the years 1994-1996 when vascular deaths numbered 554, 573 and 524 respectively. The mean age of the patients who died from vascular disease in 1997 was 73 (range 19-92) years. A consultant surgeon made the decision to operate in 98 per cent of the patients with vascular disease who died at operation and was the principal surgeon in 70 per cent of cases. Vascular consultants carried out a higher percentage of operations than their counterparts in other surgical specialties (general surgery, 63 per cent; mean for other subspecialties, 60 per cent). In the non operative group of vascular deaths in 1997, a vascular consultant was in charge in 66 per cent of cases; the figure for operated cases was 87 per cent. Adverse factors were believed to contribute to death in 10 per cent of cases, but were considered a direct cause of death in fewer than 1 per cent. CONCLUSION: These figures reflect the high quality of vascular services in Scotland, where there is a considerable consultant presence in the management of high-risk patients. This consultant involvement is higher than in other subspecialties and, bearing in mind the high percentage of emergencies, has significant resource implications for the delivery of vascular services. PMID- 10361217 TI - Vascular surgical society of great britain and ireland: RETA: the registry of endovascular treatment of abdominal aortic aneurysms AB - BACKGROUND: RETA is a voluntary registry within the UK established as a joint venture between radiologists and surgeons. The aim is to evaluate carefully the endovascular treatment of abdominal aortic aneurysms during introduction into clinical practice. METHODS: Some 367 patients have been included in the registry from 23 centres since its inception on 1 January 1996. In the majority (82 per cent) an elective repair was performed for an asymptomatic aneurysm. The mean age of the patients was 71.9 years and 28 per cent were deemed unsuitable for conventional repair. One hundred and ninety-nine bifurcated (54 per cent), 18 tube (5 per cent) and 148 aortouni-iliac with crossover (40 per cent) stent graft types were placed. RESULTS: At the end of the procedure 77 per cent of aneurysms were excluded with no complications. Twenty-one patients (6 per cent) required conversion to open repair. Forty-four (12 per cent) required additional endovascular procedures. At 30 days 87 per cent of the aneurysms were excluded or repaired. The 30-day mortality rate was 7 per cent (n = 24) with a significant improvement between 1996 and 1997, from 11 to 4 per cent. There were 25 persistent leaks (7 per cent). The death rate was related to the fitness of patient, device used and requirement for conversion. One-year follow-up data are available for 140 patients. There have been 13 deaths (9 per cent), three ruptures (2 per cent) and some form of complication in 20 per cent of cases overall. CONCLUSION: Improving results have been demonstrated for the primary procedure. At present there is a relatively high complication rate and ruptures have occurred. Rigorous continued follow-up is required. PMID- 10361218 TI - D-lactate as an early marker of intestinal ischaemia after ruptured abdominal aortic aneurysm repair. PMID- 10361219 TI - Repair of ventral hernias with expanded polytetrafluoroethylene patch Author's reply PMID- 10361220 TI - Water-soluble contrast study predicts the need for early surgery in adhesive small bowel obstruction. PMID- 10361221 TI - Water-soluble contrast study predicts the need for early surgery in adhesive small bowel obstruction Authors' reply PMID- 10361222 TI - Laparoscopic subtotal cholecystectomy in patients with complicated acute cholecystitis or fibrosis Authors' reply PMID- 10361223 TI - The role of apoptosis in the initiation of the autoimmune response in Sjogren's syndrome. PMID- 10361224 TI - Studies of the mechanism of cytolysis by tumour-infiltrating lymphocytes. AB - In order to determine the mechanism of tumour destruction by tumour-infiltrating lymphocytes (TIL), we examined the ability of both CD4+ and CD8+ effector TIL, and TIL clones, to manifest granzyme-mediated and Fas-mediated destruction of tumour targets. In many in vitro studies TIL have been shown to manifest anti tumour reactivity, yet many tumours escape immunological destruction. To investigate the role of Fas expression and the concomitant sensitivity to the inducibility of apoptotic death, we derived TIL from four melanomas and one glioma. The glioma, and all but one of the melanomas, expressed Fas, but Fas mediated apoptosis could only be detected if the targets were treated with cyclohexamide. The melanomas and the glioma all expressed detectable cytoplasmic Bcl-2 protein, known to exert anti-apoptotic activity. Lysis of tumours by CD8 enriched cultures and CD8+ clones was Ca2+-dependent and could not be modified by an anti-Fas MoAb. In CD4-enriched cultures or CD4+ clones with cytotoxic potential against tumour cells, cytotoxicity was also Ca2+-dependent. As Ca2+ dependent cytotoxicity is usually the result of secretion of perforin/granzyme-B, we investigated the presence of perforin in cytotoxic CD4+ clones and demonstrated the presence of granular deposits of this enzyme in some of the CD4+ clones. Although an anti-Fas MoAb did not block the lysis of melanoma targets by CD4+ clones, the examination of Fas-dependent targets demonstrated that these clones also had the potential to kill by the Fas/Fas ligand system. These data suggest that the predominant mechanism in tumour killing by TIL appears to be perforin-granzyme-dependent, and that the solid tumour cell lines we studied are less susceptible to Fas-mediated apoptosis. As non-apoptotic pathways may enhance tumour immunogenicity, exploitation of the perforin-granzyme-dependent cytotoxic T lymphocyte (CTL) pathways may be important for achieving successful anti-tumour responses. PMID- 10361225 TI - Sex-limited protein: in vitro and in vivo functions. AB - Mouse complement component C4 exists in two isoforms, C4 and a protein with expression restricted to male animals called sex-limited protein (Slp). Although Slp is about 95% homologous to C4, it is generally believed to be non-functional, at least in conventional haemolytic complement assays. In a previous study, however, we showed that Slp is haemolytically active in a C1-inhibitor (C1INH) regulated, EDTA-resistant mouse complement activation pathway. To study other possible implications of this finding, we generated constitutively expressing Slp transgenic mice. The transgene was crossed into otherwise Slp-deficient C57Bl/6J and NZB mice. Members of the third backcross generation of C57Bl/6J mice were tested for functional Slp and classical and alternative complement pathway activities (CH50 and AP50 levels, respectively). Slp-transgenic C57Bl/6J mice showed enhanced CH50, but normal AP50 levels when compared with non-transgenic littermates. To discover a possible protective role for Slp in spontaneous systemic lupus erythematosus (SLE) in NZBxNZW (NZBxW) mice, the third backcross generation of Slp-transgenic NZB mice was mated with NZW mice and the development of SLE in the female offspring was followed. In these introductory experiments, Slp-transgenic NZBxW animals presented with a significantly extended life span. Our results imply that Slp is a mouse complement component with functions which partially resemble some of those of human C4A. PMID- 10361226 TI - Complement activation and expression of membrane regulators in the middle ear mucosa in otitis media with effusion. AB - The aetiopathogenesis of chronic otitis media with effusion (OME) in children is not yet fully understood. OME is characterized by metaplasia of the epithelium and accumulation of sticky, glue-like effusion in the middle ear containing different mediators of inflammation, including activation fragments of the complement system. Here we examined whether the fluid phase complement activation is reflected in the middle ear mucosa and how the mucosa is protected against the cytolytic activity of complement. Mucosal biopsies from 18 middle ears of children with a history of chronic OME were taken. The biopsies were analysed by immunofluorescence microscopy after staining for complement fragments iC3b/C3c, C3d and C9, and regulators membrane cofactor protein (MCP; CD46), decay accelerating factor (DAF; CD55) and protectin (CD59). There was a strong staining for iC3b/C3c, and a weaker one for C3d and C9 on the surface of the middle ear epithelial cells of OME patients but not in controls without OME. MCP was expressed on the hyperplastic three to four outer cell layers of the epithelium, while CD59 was expressed throughout the middle ear mucosa. The results suggest a strong ongoing complement activation and consequent inflammation in the middle ear cavity. Unrestricted complement damage of the epithelial lining is prevented by the strong expression of MCP and CD59. PMID- 10361227 TI - Characterization of phenotype and cytokine profiles of T cell lines derived from vitreous humour in ocular inflammation in man. AB - Intermediate uveitis (IU) and Fuchs' heterochromic cyclitis (FHC) are two chronic ocular inflammatory disorders. They differ considerably in ocular morbidity, which is higher in IU. T cell lines were derived from the vitreous humour (VH) and peripheral blood (PB) of 10 patients with IU and four patients with FHC. There was a predominance of CD8+ in all the lines. However, there was a significantly higher percentage of CD4+ T cells in the T cell lines derived from VH of IU (32.0 +/- 8.6%) compared with FHC patients (19. 2 +/- 8.9%) (P = 0.04). The VH-derived T cell lines (VDTC) produced significantly higher levels of IL-2, interferon-gamma (IFN-gamma) and IL-10, but not IL-4, compared with PB-derived T cell lines (PBDTC) in both entities. There was significantly higher IL-2 production by VDTC from IU when compared with FHC patients (1810 +/- 220 pg/ml versus 518 +/- 94 pg/ml; P = 0.009), which could account for the more aggressive clinical features of this condition. In contrast IL-10 production was significantly higher by the VDTC from FHC compared with IU patients. The high IL 10 production by T cells infiltrating VH of FHC patients could down-regulate the inflammatory responses, thereby contributing to the benign clinical course seen in these patients. The accumulation of T cells with differing cytokine profiles in the VH suggests an important role for these cytokines in the pathogenesis of these chronic uveitides. PMID- 10361229 TI - Increased collagenase and dipeptidyl peptidase I activity in leucocytes from healthy elderly people. AB - The incidence of infectious diseases increases with ageing. The enzymatic activity of leucocytes may have a relevant role in the morbidity and mortality due to infections in the elderly. In this study we have compared the activity of enzymes involved in the inflammatory response in leucocytes from young and elderly women. A total of 35 healthy females was studied, 20 volunteers aged 78 98 years (mean 89.1 years) and 15 young controls aged 19-34 years (mean 26 years). All of them were in good clinical condition, without any acute or chronic disease. Intracellular enzyme activity was analysed by flow cytometry in leucocytes from young and elderly women. The enzyme substrates employed were for oxidative burst, L-aminopeptidase, collagenase, cathepsin B, C, D and, G and dipeptidyl peptidase I. The intracellular enzyme activity assessed by flow cytometry in leucocytes from young and elderly women was similar, as far as oxidative burst, L-aminopeptidase, cathepsin B, C, D and G are concerned. An increased collagenase activity was detected in granulocytes from elders. The mean fluorescence channels for this enzyme corresponded to 86 +/- 23 and 60 +/- 15 in cells from elders and controls, respectively (P = 0.01224). An increased dipeptidyl peptidase I activity was detected in lymphocytes from elderly women. The corresponding values for this enzyme in elders and the young were 65.9 +/- 43.3 and 17.3 +/- 5, respectively (P = 0. 0036). The proper functional activity of intracellular enzymes involved in inflammatory responses is likely to be determinant for successful ageing. PMID- 10361228 TI - Tumour necrosis factor-alpha (TNF-alpha) transcription and translation in the CD4+ T cell-transplanted scid mouse model of colitis. AB - The adoptive transfer of activated CD4+ alpha/beta T cell blasts from the spleens of immunocompetent C.B-17+/+ or BALB/cdm2 mice into C.B-17scid/scid (scid) mice induces a colitis in the scid recipient within 8 weeks, which progresses to severe disease within 16 weeks. T cells isolated from recipient colon show a Th1 cytokine phenotype. We have examined the relationship between the phenotype of the cellular infiltrate and the transcription and translation of the proinflammatory cytokine TNF-alpha. The techniques of double indirect immunohistology and in situ hybridization using digoxigenin-labelled riboprobes were used. The prominent myeloid cell infiltrate in diseased tissues comprised F4/80+, Mac-l+ macrophages, neutrophils, dendritic cells and activated macrophages. TNF-alpha transcription and translation were associated with activated macrophages in the lamina propria. Activated macrophages transcribing and translating TNF-alpha were clustered in areas of tissue destruction. Crypt epithelium of inflamed tissues transcribed TNF-alpha at a very early stage of the disease process, but translation of TNF-alpha protein could only be found in advanced epithelial dysplasia. This indicates differential post-transcriptional control of TNF-alpha in activated macrophages and the epithelium. PMID- 10361230 TI - B and T cell immunity in patients with lysinuric protein intolerance. AB - Lysinuric protein intolerance (LPI) is characterized by defective cellular transport of the dibasic amino acids, secondary dysfunction of the urea cycle, aversion to dietary protein, failure to thrive, hepatosplenomegaly and osteoporosis. Because several patients have suffered from recurrent respiratory infections and/or severe generalized varicella, and a few have developed systemic lupus, vasculitis or other autoimmune diseases, we have now evaluated the function of patients' immune systems. Serum concentrations of one to three IgG subclasses were decreased in 10 of the 12 patients studied. Antibody titres against diphtheria, tetanus and Haemophilus influenzae (Hib) were below the detection limit of the assay in four, three and eight of the 11 patients examined, respectively. (Re)vaccination of these 11 patients led to satisfactory responses against tetanus, but two patients still failed to develop measurable antibodies against diphtheria, two against Hib and six against one or more of the three serotypes of 23-valent pneumococcus vaccine. The proportions of T cells of all lymphocytes and the proliferative responses of the peripheral blood mononuclear cells were normal. In conclusion, humoral immune responses in some patients with LPI are defective and these patients may benefit from intravenous immunoglobulin therapy. PMID- 10361231 TI - Modulation of neopterin formation and tryptophan degradation by Th1- and Th2 derived cytokines in human monocytic cells. AB - In order to examine the regulatory effects of major Th1-derived cytokines, such as IL-12, and Th2 cytokines, IL-4 and IL-10, on the formation of neopterin and degradation of tryptophan, two metabolic pathways induced by interferon-gamma (IFN-gamma) in human monocytes/macrophages, we investigated the human monocytic cell line THP-1, primary human macrophages, and peripheral blood mononuclear cells (PBMC). Neopterin formation and tryptophan degradation were induced similarly by IFN-gamma in all three cell types investigated, but the effects of interleukins were different between THP-1, primary macrophages and PBMC. In PBMC, but not in THP-1 cells and primary macrophages, IL-12 was found to be additive to the effects of IFN-gamma to superinduce neopterin formation and tryptophan degradation. IL-4 and IL-10 reduced the effects of IFN-gamma on monocytic cells, and both cytokines were additively antagonistic to IFN-gamma in PBMC and THP-1 cells. Finally, on preincubation, but not on addition of IL-12, the effects of IL 4 and IL-10 on PBMC could be abrogated, whereas no such effect was seen in THP-1 cells. The results show that IL-12 up-regulates neopterin formation and tryptophan degradation by inducing additional IFN-gamma production by Th1 cells, while a direct effect of IL-12 on monocytes/macrophages appears to be absent. Similarly, IL-4 and IL-10 inhibit neopterin production and tryptophan degradation in PBMC by down-regulating Th1-type cytokine production and possibly also via direct deactivation of IFN-gamma effects towards monocytes/macrophages. The results clearly show how Th1 cell-mediated immunity may be up- or down-regulated by endogenous cytokine production. PMID- 10361233 TI - Role of decidual natural killer (NK) cells in patients with missed abortion: differences between cases with normal and abnormal chromosome. AB - In order to study the mechanism of abortion, the proportions of NK cells in the peripheral blood and decidual lymphocytes were evaluated in both chromosomally normal and abnormal missed abortions. In normal pregnancy, CD56+16-3- NK cells are a major element of decidual lymphocytes. The percentages of CD56+16-3-NK cells of peripheral lymphocytes in normal pregnancies were not statistically significantly different from those of chromosomally normal and abnormal abortions. In the decidua, the percentages of CD56+16-3- NK cells of decidual lymphocytes showed no statistically significant differences between normal pregnancies and chromosomally abnormal abortions. However, the percentages of CD56+16-3-NK cells of chromosomally normal abortions were lower than those of chromosomally abnormal (P = 0.0025). Moreover, the percentages of CD56+16- NK cells in abortions with normal chromosomes were lower than those in normal pregnancies or abortions with abnormal chromosomes (P = 0.0037, P = 0.0025). However, when the proportion of CD56+ NK cells expressing CD16 was evaluated, there were no statistically significant differences in the percentages of CD56+16+ NK cells in normal pregnancies and missed abortions with normal chromosomes and abnormal chromosomes. We conclude that the expression of decidual CD56+16-3- NK cells in missed abortions with normal chromosomes is different from abortions with abnormal chromosomes and that this phenomenon may depend on an abnormal immune response of the maternal side. PMID- 10361232 TI - Human B cells accumulate immunoglobulin V gene somatic mutations in a cell contact-dependent manner in cultures supported by activated T cells but not in cultures supported by CD40 ligand. AB - The acquisition of somatic mutations in the rearranged immunoglobulin V regions in B cells occurs within the tightly regulated microenvironment of a germinal centre. The precise mechanism responsible for turning on the mutational process is unknown. To dissect the role of different components of the germinal centre in this mechanism, we have used in vitro cultures of normal human IgD+ peripheral blood B lymphocytes co-cultured with activated CD4+ T cells, or with resting CD4+ T cells, or with CD40 ligand and IL-4. We observed that if the cultures included activated CD4+ T cells, then up to 100% of VH transcripts on day 14 were somatically mutated. Transcripts were found to carry from one to 36 substitutions (median five). In contrast, in the absence of activated T cells, transcripts contained only background levels of somatic mutation irrespective of the presence of resting T cells or CD40 ligand and IL-4. Cell-cell contact was required for mutation because mutations were not detected when B cells were separated from activated T cells by a membrane. PMID- 10361234 TI - Porins and lipopolysaccharide (LPS) from Salmonella typhimurium induce leucocyte transmigration through human endothelial cells in vitro. AB - Bacteria or bacterial products may constitute important inducers of surface molecule expression on endothelial cells and leucocytes. This study was undertaken to determine the effects of the Salmonella typhimurium porins, LPS-S and LPS-R on the transendothelial migration of leucocytes through human umbilical vein endothelial cells (HUVEC). Treatment of the HUVEC with either porins or LPS S or LPS-R increased the transmigration of different leucocyte populations, in particular that of neutrophils. The maximal increase occurred using LPS-S treatment, whereas porin stimulation fell between LPS-S and LPS-R. The transmigration increase was dose-dependent and reached its maximum at about 100 1000 ng/ml of stimulus. Optimal endothelial activation occurred after 2-4 h and 4 6 h using LPS and porin, respectively. Stimulation of leucocytes with either porins or LPS slightly increased their transmigration through non-activated endothelial cells. Transmigration increased remarkably during the simultaneous stimulation of endothelial cells by IL-1ss together with either porins or LPS. To assess participation of E-selectin, intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and leucocyte adhesion complex (CD11/18) in porin- or LPS-mediated leucocyte migration, blocking MoAbs were used. Each blocking MoAb partially and selectively decreased leucocyte transmigration. The obtained results contribute to clarify some aspects of the inflammatory process at sites of infection. PMID- 10361235 TI - Depletion of liver and splenic macrophages reduces the lethality of Shiga toxin-2 in a mouse model. AB - The haemolytic uraemic syndrome (HUS) is a clinical syndrome consisting of haemolytic anaemia, thrombocytopenia, and acute renal insufficiency. HUS is the most frequent cause of acute renal failure in childhood. It has been previously suggested that the presence of Shiga toxin (Stx) is necessary but not sufficient for HUS development, and cytokines such as tumour necrosis factor-alpha (TNF alpha) and IL-1beta appear to be necessary to develop the syndrome. Since the mononuclear phagocytic system (MPS) is the major source of these cytokines, macrophages might be one of the relevant targets for Stx action in the pathophysiology of HUS. In this study our objective was to examine the role of the hepatic and splenic macrophages in a mouse model of HUS induced by injection of Shiga toxin type-2 (Stx2) or Stx2 plus lipopolysaccharide (LPS). For this purpose, depletion of mice macrophages by liposome-encapsulated clodronate (lip clod), followed by injection of STx2 or Stx2 plus LPS, was assayed. In this study we show that depletion of hepatic and splenic macrophages by clodronate treatment induces a survival of 50% in animals treated with Stx2 alone or in presence of LPS. This maximal effect was observed when lip-clod was injected 48-72 h before Stx2 injection. Biochemical and histological parameters show characteristics of the lesion produced by Stx2, discarding non-specific damage due to LPS or lip clod. In addition, we determined that the toxic action of Stx2 is similar in BALB/c and N:NIH nude mice, indicating the T cell compartment is not involved in the Stx2 toxicity. Briefly, we demonstrate that macrophages play a central role in the pathophysiology of HUS, and that the systemic production of cytokines by liver and/or spleen is for Stx2 to manifest its full cytotoxic effect. In addition, the toxicity of Stx2 alone, or in presence of LPS, is independent of the T cell compartment. PMID- 10361236 TI - Analysis of Epstein-Barr virus (EBV) receptor CD21 on peripheral B lymphocytes of long-term EBV- adults. AB - Primary infections with EBV are rarely observed after the age of 20. Some individuals even remain seronegative all their lives. Previously, a lack of EBV receptors on B cells of persistently EBV- adults was described as a reason for long-term EBV-seronegativity. The present study examined the CD21 receptor status of 20 repeatedly EBV- healthy adults and 32 EBV+ volunteers by means of flow cytometry. CD21 molecules on the surface of CD19+ B cells were quantified using anti-IgG-coated microbeads. The percentage of CD19+/CD21+ B lymphocytes was slightly lower in the peripheral blood of EBV- donors, but the CD21 antibody binding capacity on CD19+ B cells showed no significant differences between EBV- and EBV+ adults. In vitro studies showed an equally good EBV transformability of peripheral B lymphocytes of EBV- and EBV+ donors. Since HLA-DR was recently described as a co-receptor for EBV infection of B cells, we also determined HLA DRB1 alleles in the EBV- group. We found a significant negative association of EBV-seronegativity with HLA-DR13 in comparison with 111 healthy blood donors. In summary, a biologically significant lack of the EBV receptor CD21 on peripheral B lymphocytes of persistently EBV- adults was excluded as a reason for long-term EBV-seronegativity. PMID- 10361237 TI - Proviral load and immune function in blood and lymph node during HIV-1 and HIV-2 infection. AB - Proviral load as well as lymphocyte phenotype and function were compared in peripheral blood and lymph node compartments of 17 HIV-1, 12 HIV-2 and three dually infected patients with lymphadenopathy. The mean percentage (95% confidence interval (CI)) of CD4+ cells was higher in lymph node mononuclear cells (LNMC) than in peripheral blood mononuclear cells (PBMC) in both infections, being 26.7% (21. 1%, 32.3%) and 15.3% (10.4%, 20.2%), respectively, for HIV-1-infected patients (P = 0.0001) and 32.3% (22.7%, 41.9%) and 22. 1% (13.6%, 30.6%), respectively, for HIV-2-infected patients (P = 0. 02). In both types of infection, proviral load adjusted for number of CD4+ cells was higher in LNMC than in PBMC: the geometric mean (95% CI) was 8937 (4991; 16 003) and 4384 (2260; 8503), respectively, for HIV-1 patients (P = 0.02) and 1624 (382; 6898) and 551 (147; 2058) DNA copies, respectively, for HIV-2 patients (P = 0.05). Proviral load in both compartments was closely correlated (HIV-1, r = 0.60, P = 0.01; and HIV-2, r = 0.83, P = 0.0003). In both infections, proliferation and interferon-gamma (IFN-gamma) production in response to purified protein derivative (PPD) was lower in LNMC than in PBMC, both of which, in turn, were lower than in healthy controls. These results indicate that in HIV-2 as in HIV-1 infection, infected cells have a tropism for the lymph nodes resulting in higher viral load in this compartment and lower lymphocyte responses to the recall antigen PPD which may increase susceptibility to tuberculosis. PMID- 10361238 TI - CD8+ T cell-mediated enhancement of tumour necrosis factor-alpha (TNF-alpha) production and HIV-1 LTR-driven gene expression in human monocytic cells is pertussis toxin-sensitive. AB - HIV replication and LTR-mediated gene expression can be modulated by CD8+ T cells in a cell type-dependent manner. We have previously shown that supernatants of activated CD8+ T cells of HIV-infected individuals greatly enhanced p24 levels in human macrophages infected with NSI or SI primary isolates of HIV-1. Here we have examined the effect of culture with CD8+ T cell supernatants on HIV-1 LTR mediated gene expression in monocytic cells. CD8+ T cell supernatants enhanced LTR-mediated gene expression in U38 cells activated with Tat in the absence or presence of phorbol myristate acetate (PMA) and ionomycin or TNF-alpha. Further, enhancement of LTR-mediated gene expression and virus replication in U38 cells and U1 cells, respectively, was pertussis toxin-sensitive. The enhancement of gene expression and virus replication was associated with increased levels of TNF alpha and was significantly abrogated by antibody to TNF-alpha. In contrast, the suppression of LTR-mediated gene expression by CD8+ T cell supernatants in Jurkat T cells was not pertussis toxin-sensitive and TNF-alpha levels were not affected. These results demonstrate that factors produced by CD8+ T cells utilize different cellular pathways to mediate their effects on HIV transcription and replication in different cell types. PMID- 10361239 TI - Immunological changes in peripheral blood and in lymphoid tissue after treatment of HIV-infected subjects with highly active anti-retroviral therapy (HAART) or HAART + IL-2. AB - This study presents the immunophenotypic and functional analysis of lymphocyte subsets obtained from peripheral blood and lymphoid tissue from HIV+ individuals treated with highly active anti-retroviral therapy (HAART) alone or in combination with 6 million units international (MUI) s.c. IL-2. Before treatment, the HIV+ patients had reduced CD4 and increased CD8 values in the peripheral blood and lymphoid tissue and impaired cytokine production by peripheral blood mononuclear cells (PBMC). After 24 weeks of treatment, all the HIV+ patients demonstrated increased CD4 values in peripheral blood and lymphoid tissue. The use of IL-2 did not promote an additional CD4 expansion compared with HAART alone; increased 'naive' and CD26+ CD4 cells and reduced CD8 cells were found in the peripheral blood and lymphoid tissue of the IL-2-treated, but not of the HAART-treated patients. Both types of treatment induced a significant reduction of the CD8/CD38+ cells. While HAART alone had negligible effects on cytokine production by PBMC, the combined use of HAART + IL-2 was unable to increase the endogenous production of IL-2, but caused an increase of IL-4, IL-13 and interferon-gamma (IFN-gamma) and a reduction of monocyte chemoattractant protein 1 (MCP-1) production. These data suggest that, although in this schedule IL-2 has minimal efficacy on CD4 recovery when compared with HAART alone, it produces an increase of 'naive' and CD26+ CD4 cells and a partial restoration of cytokine production. These data may be used to better define clinical trials aiming to improve the IL-2-dependent immunological reconstitution of HIV-infected subjects. PMID- 10361240 TI - Antibodies to a non-repeat region of Plasmodium falciparum antigen Pf155/RESA in individuals from malaria-endemic areas. AB - Human antibodies to the repeat regions of the Plasmodium falciparum asexual blood stage antigen Pf155/RESA interfere with parasite growth in vitro, but the significance in this respect of antibodies to non-repetitive epitopes is less clear. In this study the levels of antibodies to a non-repetitive part of Pf155/RESA (residue 199-221) in malaria-exposed individuals were analysed, as was the parasite-inhibitory capacity of such antibodies. Residue 199-221 is of particular interest since it includes a sequence homologous to a cytoadherence related motif from band 3. Sera from donors in Liberia and Tanzania were analysed for reactivity in ELISA with synthetic peptides together overlapping this part of Pf155/RESA. High antibody reactivity was observed in most of the sera with two peptides including residues 199-211 and 202-214, respectively. Specific antibodies were affinity-purified from selected sera using these peptide sequences and were shown to react with Pf155/RESA by immunofluorescence and Western blotting. The purified antibodies were furthermore shown to inhibit parasite growth in vitro. The results suggest that both repeat and non-repeat epitopes in Pf155/RESA elicit antibodies with potential to protect against malaria infection. PMID- 10361242 TI - The significance of IgG subclasses and mannan-binding lectin (MBL) for susceptibility to infection in apparently healthy adults with IgA deficiency. AB - The aim of this study was to investigate the significance of IgG subclasses and MBL for susceptibility to infection in association with IgA deficiency. The study population consisted of 139 apparently healthy adult blood donors with IgA deficiency and normal serum levels of IgG and IgM, and an increased susceptibility to infection demonstrated at a population level. Additionally, 216 controls matched for age and sex were investigated. IgG4 deficiency was more common and the mean level of IgG4 lower in persons with IgA deficiency than in the controls. No significant associations could be demonstrated between overt IgG subclass deficiencies and increased susceptibility to infection. However, when the mean concentrations of IgG subclasses were analysed with regard to medical history, that of IgG1 was lower in persons who reported recurrent viral respiratory infections, that of IgG3 in persons who had episodes of severe infection in their history, and that of IgG4 in persons who had recurrent mild respiratory infections, compared with those who had no particular history of infections. In contrast, MBL deficiency-alone or combined with that of the IgG subclass-was not associated with increased susceptibility to infection in persons with IgA deficiency. The results indicate that the proneness to infections observed in a population of otherwise healthy persons with IgA deficiency can only for a small part be accounted for by concomitant deficiencies of IgG subclasses. Contrary to expectations, no synergism between the deficiencies of IgA and MBL could be demonstrated. PMID- 10361241 TI - Leishmania-specific T cells expressing interferon-gamma (IFN-gamma) and IL-10 upon activation are expanded in individuals cured of visceral leishmaniasis. AB - Peripheral blood mononuclear cells (PBMC) from patients who have recovered from visceral leishmaniasis often respond to Leishmania antigens in vitro by production of both IL-4, IFN-gamma and IL-10. In order to establish the cellular sources of these cytokines, we activated cells from individuals with a history of visceral leishmaniasis with Leishmania antigen for 6 days in culture, and identified cytokine production at the single-cell level by flow cytometry. The cytokines were only found in CD3+ cells and among these mainly within the CD4+ subset. The percentage of cytokine-producing cells was compared in Leishmania activated PBMC cultures from the previous patients and from individuals living in a village where leishmaniasis does not occur. The percentage of IL-10- and IFN gamma-containing cells was significantly higher in the previous patients than in the controls, indicating that Leishmania-specific T cells producing IL-10 and/or IFN-gamma had been expanded as a result of the infection. The cytokine-producing cells in the previous patients could be divided into three types: (i) cells producing IFN-gamma only; (ii) cells producing IL-4 only; and (iii) cells producing IFN-gamma and IL-10 simultaneously. The first and second group of cells can be described as Th1- and Th2-type cells, respectively. The third group could be a regulatory subset of T cells important for maintaining a balance between Th1 and Th2-type cells in these individuals. PMID- 10361243 TI - In vivo modulation of cytokine synthesis by intravenous immunoglobulin. AB - We examined the effects of intravenous immunoglobulin (IVIG) on cytokine regulation in vivo using samples taken before and after replacement-dose (200-400 mg/kg) IVIG in a group of patients with common variable immunodeficiency (CVID) and X-linked agammaglobulinaemia (XLA). The intracellular cytokine content of CD4+ and CD8+ lymphocytes, and their CD28+/- subsets, were measured following in vitro activation with phorbol myristate acetate (PMA) and ionomycin. The cytokines IL-2, interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha), and the early activation marker CD69, were assessed by four-colour flow cytometry of whole blood cultures taken before and after IVIG infusion. There was a significant increase in IL-2 expression in CD4+ (and CD4+28-) cells and an increase in TNF-alpha expression in CD8+28- cells following IVIG in CVID, but not in XLA patients. IFN-gamma and CD69 expression were not affected by IVIG infusion. This increase in TNF-alpha and IL-2, combined with unchanged IFN-gamma expression, is evidence against the putative 'anti-inflammatory' role of IVIG, and may explain the failure of resolution of granulomata in CVID patients treated with IVIG alone. PMID- 10361244 TI - HLA class II homozygosity confers susceptibility to common variable immunodeficiency (CVID). AB - Most cases of CVID occur sporadically, but familial cases do also occur and 15% of the patients with the disease have first degree relatives with IgA deficiency (IgAD). Our purpose was to study CVID association with HLA class II alleles and to ascertain whether this disease shares a common genetic background with IgAD in our population. Patients with CVID (n = 42), were typed using gene amplification and sequence-specific oligonucleotide probing for HLA-DRB1, DRB3, DQA1 and DQB1 loci and their typing compared with that of 96 IgAD and 334 healthy controls. We observed a positive association between non-Asp residues at position 57 of the HLA-DQbeta chain and CVID, although much weaker than in IgAD. Further, we found an association between CVID and homozygosity for genes encoding HLA class II molecules, especially HLA-DQ, not seen in IgAD. The data support the hypothesis that a restricted diversity of HLA class II molecules may contribute to susceptibility to CVID. PMID- 10361245 TI - Mode of delivery directs the phagocyte functions of infants for the first 6 months of life. AB - Factors that direct the immune responsiveness of the newborn beyond the immediate post-natal period are not known. We investigated the influence of mode of delivery and type of feeding on the phagocyte activity during the first 6 months of life. Sixty-four healthy infants (34 delivered vaginally and 30 by elective Caesarean section) were studied at birth and at the ages of 2 and 6 months. Phagocyte functions were studied by measuring the chemiluminescence (CL) activity of whole blood and isolated leucocytes and by investigating the expression of phagocyte receptors (FcgammaRI (CD64), FcgammaRII (CD32), FcgammaRIII (CD16), CR1 (CD35), CR3 (CD11b) and FcalphaR (CD89)) on neutrophils, monocytes and eosinophils by using receptor-specific MoAbs and immunofluorescence flow cytometry. Infants born by elective Caesarean section had significantly higher CL activity than those delivered vaginally during the entire 6-month follow up. In addition, infants who received formula feeds had significantly higher CL activity at 6 months of age and higher expression of FcgammaRI-, Fcalpha- and CR3 receptors on neutrophils than infants exclusively breast-fed. We suggest that stress reaction associated with labour influences the phagocytic activity measured in the cord blood but later during infancy the intraluminal antigens, gut microflora and diet, become important determinants in immune programming of human individuals. PMID- 10361246 TI - Rapid conversion of naive to effector T cell function counteracts diminished primary human newborn T cell responses. AB - The reduced incidence of graft versus host disease following the use of human cord blood as a source of stem cells for bone marrow reconstitution challenges our understanding of the immunocompetence of newborn T cells. Newborn CD4+ T cells express mainly the CD45RA phenotype and have been considered to respond comparably to adult CD4+ T cells exhibiting the CD45RA phenotype. We compared the in vitro kinetics of phenotypic conversion of newborn and adult CD4+CD45RA+ T cells to CD4+CD45RO+ T cells. The cytokine profile and B cell helper activity of the converted CD4+CD45RO+ T cell population were also determined. Newborn CD4+CD45RA+ T cells were converted to CD4+CD45RO+ with significantly faster time kinetics than adult CD4+CD45RA+ T cells, following either phytohaemagglutinin (PHA) or anti-CD2 activation. Freshly purified newborn naive T cells did not produce IL-2, IL-4 or interferon-gamma (IFN-gamma) following stimulation, whereas adult naive T cells secreted IL-2 and adult-derived CD4+CD45RO+ T cells secreted all three cytokines under the same stimulatory conditions. However, newborn and adult CD4+CD45RA+ T cells, following primary stimulation and maturation in vitro, acquired the ability to secrete a Th1-type cytokine profile of IL-2 and IFN-gamma after secondary stimulation. Newborn CD4+ naive T cells that acquired the CD45RO phenotype in vitro also gained B cell helper activity equivalent to that of adult in vitro matured CD4+ naive T cells. These findings suggest that newborn and adult CD4+CD45RA+ T cell subsets are differentially responsive to various stimuli. They show that newborn CD4+CD45RA+ naive T cells can transform more quickly than their adult counterparts into functionally equivalent CD4+CD45RO+ T cells, a process that may be important to counteract the immature immune environment which exists in the newborn. PMID- 10361248 TI - Elevated serum levels of soluble membrane cofactor protein (CD46, MCP) in patients with systemic lupus erythematosus (SLE). AB - Membrane cofactor protein (MCP, CD46) is a cell surface complement regulatory protein which acts as a cofactor for the factor I-mediated cleavage of the activated complement components C3b/C4b. To evaluate the clinical usefulness of serum soluble CD46 as a marker of disease activity in patients with SLE, serum levels of sCD46 were measured by ELISA, using two MoAbs (M160 and M177), each of which recognized two different epitopes on CD46 molecule in SLE, other autoimmune diseases and healthy controls. Serum sCD46 levels in active SLE patients (30.5 +/ 14.1 ng/ml) were significantly higher than those of inactive SLE (5.8 +/- 7.1 ng/ml; P = 0.0003), rheumatoid arthritis (14.9 +/- 11.6 ng/ml; P = 0.0218), primary Sjogren's syndrome (12.3 +/- 11.6 ng/ml; P = 0.0039) and normal controls (7.3 +/- 3.6 ng/ml; P = 0.0005). The elevated serum sCD46 levels in active SLE patients significantly decreased from 30.5 +/- 14.1 ng/ml to 8.0 +/- 6.3 ng/ml after effective corticosteroid and immunosuppressant therapy (P = 0.018). Additionally, we found a significant negative association between increasing concentration of sCD46 and decreasing levels of CH50 in SLE (r = -0.598, P = 0.0009). These results suggest that sCD46 reflects in vivo activation of complement system and provides an additional useful serum parameter of active SLE. PMID- 10361247 TI - Beneficial effect of the inosine monophosphate dehydrogenase inhibitor mycophenolate mofetil on survival and severity of glomerulonephritis in systemic lupus erythematosus (SLE)-prone MRLlpr/lpr mice. AB - The aim of the present study was to evaluate the therapeutic effect of mycophenolate mofetil (MMF) on the course of disease in SLE-prone MRLlpr/lpr mice. Three-months-old mice displaying clinical symptoms of glomerulonephritis were given MMF (100 mg/kg per day) orally via the drinking water. Control mice received i.p. injections of cyclophosphamide (CYC) (1.8 mg/mouse per week) or saline. Survival, albuminuria and haematuria, immunoglobulin levels and anti dsDNA antibodies in serum, frequencies of immunoglobulin-producing B lymphocytes and glomerular deposits of immunoglobulin and C3 were analysed. The results showed that MMF treatment significantly prolonged survival and reduced the occurrence of albuminuria and haematuria in MRLlpr/lpr mice. In addition, the number of immunoglobulin-producing B cells and serum levels of IgG and IgG anti dsDNA antibodies were reduced after MMF and CYC treatment. MMF treatment significantly reduced the extent of deposition of C3 in glomeruli. We conclude that the reduced severity of glomerulonephritis following treatment of lupus prone mice with MMF was as efficacious as that of CYC. These results warrant clinical trials of MMF in SLE patients with glomerulonephritis. PMID- 10361249 TI - Antibody-independent classical complement pathway activation and homologous C3 deposition in xeroderma pigmentosum cell lines. AB - Of human malignantly transformed cell lines, xeroderma pigmentosum (XP) cell lines were found to be highly susceptible to homologous complement (C): cells were opsonized by C3 fragments on incubation with diluted normal human serum. C3 fragment deposition on XP cells was Ca2+-dependent and occurred on live cells but not UV-irradiated apoptotic cells. (Ca2+ is required for activation of the classical C pathway via C1q and the lactin pathway via mannose binding lectin (MBL), and the surface of apoptotic cells usually activates the alternative C pathway.) In this study we tested which of the pathways participates in XP cell C3 deposition. In seven cell lines that allowed C3 deposition (i), Clq was shown to be essential but MBL played no role in C activation, (ii) Cls but not MASP bound XP cells for activation, (iii) no antibodies recognizing XP cells were required for homologous C3 deposition, and (iv) the alternative pathway barely participated in C3 deposition. Furthermore, the levels of C-regulatory proteins for host cell protection against C, decay-accelerating factor (DAF, CD55) and membrane cofactor protein (MCP, CD46), were found to be relatively low in almost all XP cell lines compared with normal cells. These results indicate that XP cells activate the classical C pathway in an antibody-independent manner through the expression of a molecule which directly attracts C1q in a C-activating form, and that relatively low levels of DAF and MCP on XP cells facilitate effective C3 deposition. The possible relationship between the pathogenesis of XP and our findings is discussed. PMID- 10361251 TI - Involvement of ribosomal proteins in regulating cell growth and apoptosis: translational modulation or recruitment for extraribosomal activity? AB - Gene recruitment is a mechanism of molecular evolution whereby a gene product can function in more than one distinct capacity. The 'one gene-dual function' phenomenon is well illustrated by crystallins, structural proteins that play both specialized roles in the eye lens and also 'housekeeping' enzyme roles. Ribosomal proteins are integral components of the basal cellular machinery involved in protein synthesis, whose roles have been regarded collectively as important, but individually somewhat mundane. However, various individual ribosomal proteins and also translation initiation and elongation factors have been found to play roles in regulating cell growth, transformation and death, giving rise to increasing speculation that components of the translational apparatus can act as multifunctional proteins. Recently, we have shown that ribosomal protein S3a (RPS3a) plays important roles in cell transformation and death, whereby constitutively or transiently enhanced RPS3a expression can be regarded as 'priming' a cell for apoptosis and suppression of such enhanced expression as 'execution'. While it is unclear whether RPS3a acts in a capacity mechanistically distinct from that in translation, such a possibility is discussed in this article in the light of recent, although not exhaustively reviewed, findings implicating the involvement of other individual ribosomal proteins in modulating and/or effecting changes in cellular responses and growth patterns in an extraribosomal capacity independent of their conventional role in translation. PMID- 10361250 TI - Decreased interferon-gamma (IFN-gamma)-producing T cells in patients with acute Kawasaki disease. AB - Kawasaki disease (KD) is an acute febrile illness of early childhood, in which the activation of monocytes/macrophages plays a central role in the development of vasculitis during the acute stage of disease. In this study we investigated peripheral blood T cells of 10 patients with KD, focusing on the Th1 and Th2 imbalance, using intracellular cytokine staining and analysis of the cytokine producing T cells by flow cytometry. We observed a decrease in the numbers of IFN gamma-producing, but not IL-4-producing, CD3+ T cells, during the acute stage. Our results suggest that there is an imbalance of Th1 and Th2 subsets during the acute stage of KD. PMID- 10361252 TI - Perforin-dependent nuclear targeting of granzymes: A central role in the nuclear events of granule-exocytosis-mediated apoptosis? AB - Programmed cell death, apoptosis, involves very distinctive changes within the target cell nucleus, including margination of the chromatin, DNA fragmentation and breakdown of the nuclear envelope. Cytolytic granule-mediated target cell apoptosis is effected, in part, through synergistic action of the membrane-acting protein perforin and serine proteases, such as granzymes A or B. Recent work using confocal laser scanning microscopy as well as other techniques supports the idea that perforin-dependent translocation of granzymes to the nucleus of target cells plays a central role in effecting the nuclear changes associated with apoptosis. In vitro experiments indicate that granzyme nuclear import follows a novel pathway, being independent of ATP, not inhibitable by non-hydrolysable GTP analogues and involving binding within the nucleus, unlike conventional signal- dependent nuclear protein import. In intact cells, perforin-dependent nuclear entry of granzymes precedes the nuclear events of apoptosis such as DNA fragmentation and nuclear envelope breakdown; prevention of granzyme nuclear translocation through bcl2 overexpression or treatment of target cells with inhibitors of caspase activation blocks these events. Nuclear localization of granzymes thus appears to be central to induction of the nuclear changes associated with cytolytic granule-mediated apoptosis. PMID- 10361253 TI - Activation-induced apoptosis of autoreactive and alloreactive T lymphocytes in the target organ as a major mechanism of tolerance. AB - Normal individuals have mature T lymphocytes that are capable of reacting to self antigens and can be activated by cross-reacting environmental antigens. The mechanism that maintains immune tolerance and prevents these activated autoreactive T cells from causing autoimmune disease is unclear. We have previously hypothesized that activation-induced apoptosis of previously activated autoreactive T cells in the target organ is a major mechanism for maintaining tolerance. Here I review the current evidence to support this hypothesis. It is proposed that when activated autoreactive T cells enter the target organ, they are reactivated mainly by non-professional antigen-presenting cells (APC) and deleted by activation-induced apoptosis through the Fas (CD95) pathway before producing significant target organ damage. This apoptosis occurs because the reactivated T cells do not receive sufficient costimulation from the non professional APC to up-regulate their expression of Bcl-2-related anti-apoptotic proteins, which inhibit the CD95 pro-apoptotic pathway. This is in contrast to the situation in peripheral lymphoid organs, where reactivation of T cells by professional APC results in sufficient costimulation-induced up-regulation of Bcl 2-related proteins to inhibit the CD95 pathway and allow T cell proliferation and survival as memory T cells. Activation-induced apoptosis of alloreactive T cells in allografts can similarly account for spontaneous allograft acceptance, as occurs after MHC-mismatched liver transplantation. PMID- 10361254 TI - Exposure of N-acetylglucosamine decreases early in dexamethasone-induced apoptosis in thymocytes, demonstrated by flow cytometry using wheat germ agglutinin and pokeweed mitogen. AB - In the present paper we describe changes in the exposure of oligosaccharides containing N-acetylglucosamine (Glc-NAc) during apoptosis of mouse thymocytes. The structures containing this sugar were probed with fluorescein isothiocyanate labelled lectins, wheat germ agglutinin and pokeweed mitogen in flow cytometric assays. Both lectins bind to structures containing Glc-NAc. The present report describes experiments in which two different dual-staining techniques were used to simultaneously identify apoptotic cells and measure their lectin exposure. In these experiments, we observed an early and substantial decrease in the exposure of Glc-NAc-containing structures associated with the onset of apoptosis, before or simultaneously with phosphatidylserine exposure. This was followed by an increase in the exposure of Glc-NAc-containing structures after longer incubation times, when a large proportion of cells was demonstrated to have fragmented DNA. These results provide evidence for major changes in the structure of plasma membrane oligosaccharides during apoptosis. The initial decrease may be a by product of the hydrolysis of glycosphingolipids to yield ceramide for apoptotic signalling or a deliberate process related to the removal of cell adhesion signalling structures, associated with the separation of the apoptotic cell from its neighbours. The later increase in Glc-NAc-containing structures may be the result of the incorporation of internal membranes into the plasma membrane or a deliberate production of prophagocytic signals by a still-functioning Golgi apparatus. PMID- 10361255 TI - Viral modulation of the host response via crmA/SPI-2 expression. AB - Viruses have evolved numerous strategies to modulate the host response to infection. Poxviruses cause acute infections and need to replicate quickly to promote efficient transmission. Consequently, it is not surprising to learn that poxviruses encode a large number of proteins designed to target various arms of the host inflammatory response. One of the earliest described and most well studied viral modulatory proteins is crmA/SPI-2. While the biochemical targets and possible modes of action have been well characterized in vitro, the role that crmA/SPI-2 plays during natural infection is less clear. It may have effects in modulating host responses involving apoptosis and inflammation. It is important to further understand the precise mode of action of viral proteins, such as crmA/SPI-2, because this may lead to better therapeutic strategies to combat a range of inflammatory and autoimmune diseases. PMID- 10361256 TI - Rapamycin inhibits didemnin B-induced apoptosis in human HL-60 cells: evidence for the possible involvement of FK506-binding protein 25. AB - In the present paper we show that the immunosuppressant rapamycin inhibits the induction of apoptosis by didemnin B in human promyeloid HL-60 cells. The mechanism of this inhibition is investigated using FK506, which competes with rapamycin for binding to their common target FK506-binding protein (FKBP)12. The lack of competition for rapamycin-mediated inhibition of didemnin B-induced apoptosis by FK506 suggests that rapamycin inhibits apoptosis through some mechanism other than inhibition of p70 S6 kinase activation. The lack of inhibition of didemnin B-induced apoptosis by inhibitors of phosphatidylinositol 3-kinase and mitogen-activated protein (MAP) kinase kinase further supports the conclusion that rapamycin does not inhibit didemnin B-induced apoptosis through inhibition of the MAP kinase pathway. Furthermore, didemnin B-induced apoptosis is not inhibited by the inhibitors of cyclin-dependent kinase, roscovitine and olomoucine. This indicates that rapamycin does not act through inhibition of cyclin-dependent kinases. Together with the lack of competition for the effect of rapamycin by FK506, our data suggest the possible involvement of the FK506 binding protein, FKBP25, which is localized in the nucleus. This interpretation of our data gains support from the fact that didemnin B does not induce apoptosis in enucleated HL-60 cells, which supports the possible involvement of FKBP25 in the inhibition of apoptosis by rapamycin. PMID- 10361257 TI - Elevated plasminogen receptor expression occurs as a degradative phase event in cellular apoptosis. AB - Plasminogen activation (PA) is involved in a variety of extracellular proteolytic events, such as fibrinolysis, cell migration (e.g. angiogenesis, tumour cell invasion, inflammation, wound healing, bacterial invasion), ovulation, tissue remodelling and the activation of other protease classes and growth factors. These diverse roles are due to the specific localization of components of the PA system to extracellular matrices, basement membranes, fibrin and cell surfaces. We have previously reported that PA is dramatically elevated during cycloheximide (CHX)-induced apoptosis in U937 cells due to a concomitant increase in both plasminogen receptors (PLG-R; i.e. specific PLG binding) and cell-surface urokinase plasminogen activator. We now extend this study by showing that the increase in PLG-R (resulting in an increase in specific PLG binding) is a late apoptotic event coincident with propidium iodide uptake and internucleosomal DNA fragmentation but occurring after elevations in phosphatidylserine exposure. Plasminogen was also observed to dramatically increase the rate of CHX-induced apoptosis. We conclude that PA may play a role in the degradative (i.e. late stage) events of cellular apoptosis. PMID- 10361258 TI - The IGF axis and programmed cell death. AB - Insulin-like growth factors (IGF) are mitogenic peptides that have been implicated as positive regulators of cellular proliferation. In recent years, several studies have suggested an additional role for the IGF axis in the regulation of apoptosis. Signalling through the IGF receptor has been shown to have a potent survival function and protect cells from a variety of apoptotic stimuli. The actions of IGF are regulated by a family of high-affinity IGF binding proteins (IGFBP), which sequester the IGF from the IGF receptor. However, there is some evidence that one of these binding proteins, IGFBP-3, may have its own pro-apoptotic effects that are independent of its ability to modulate IGF bioavailability. In addition, it has been suggested that the tumour suppressor p53, a crucial mediator of apoptosis in response to cellular stress, may elicit several of its apoptotic effects through manipulation of components of the IGF axis. This review summarizes what is currently known about the role of the IGF system in the regulation of apoptosis, highlighting its implications in the context of tumorigenesis. PMID- 10361259 TI - Molecular mechanisms of ionizing radiation-induced apoptosis. AB - Ionizing radiation activates not only signalling pathways in the nucleus as a result of DNA damage, but also signalling pathways initiated at the level of the plasma membrane. Proteins involved in DNA damage recognition include poly(ADP ribose) polymerase (PARP), DNA-dependent protein kinase, p53 and ataxia- telangiectasia mutated (ATM). Many of these proteins are inactivated by caspases during the execution phase of apoptosis. Signalling pathways outside the nucleus involve tyrosine kinases such as stress-activated protein kinase (SAPK)/c-Jun N terminal kinase (JNK), protein kinase C, ceramide and reactive oxygen species. Recent evidence shows that tumour cells resistant to ionizing radiation-induced apoptosis have defective ceramide signalling. How these signalling pathways converge to activate the caspases is presently unknown, although in some cell types a role for calpain has been suggested. PMID- 10361260 TI - Regulation of caspase activation and apoptosis by cellular zinc fluxes and zinc deprivation: A review. AB - Non-toxic agents that target intracellular signalling pathways in apoptosis may have potential therapeutic use in many diseases. One such agent is the transition metal Zn, a dietary cytoprotectant and anti-oxidant, which stimulates cell proliferation and suppresses apoptosis. Zn is maintained in discrete subcellular pools that are critical for the functional and structural integrity of cells. The present review initially describes the current state of knowledge on the cellular biology of Zn, especially the critical free or loosely bound (labile) pools of Zn, which are thought to regulate apoptosis. We then review the evidence relating Zn to apoptosis, including studies from our laboratory showing potent synergy between intracellular Zn deficiency and the short chain fatty acid butyrate in induction of caspase activation and the downstream events of apoptosis. Our studies have also reported the suppressive effects of micromolar concentrations of Zn on caspase-3 activation in cell-free models. Other key issues that will be discussed include the identification of the putative molecular targets of Zn and the evidence that systemic changes in labile Zn levels are sufficient to alter susceptibility to apoptosis and lead to physiopathological changes in the human body. Finally, we propose that labile Zn may serve as a coordinate regulator of mitosis and apoptosis to regulate tissue growth. PMID- 10361261 TI - Bcl-2 genes and growth factors in the pathology of ischaemic acute renal failure. AB - For the past decade, an attempt has been made by many research groups to define the roles of the growing number of Bcl-2 gene family proteins in the apoptotic process. The Bcl-2 family consists of pro-apoptotic (or cell death) and anti apoptotic (or cell survival) genes and it is the balance in expression between these gene lineages that may determine the death or survival of a cell. The majority of studies have analysed the role/s of the Bcl-2 genes in cancer development. Equally important is their role in normal tissue development, homeostasis and non-cancer disease states. Bcl-2 is crucial for normal development in the kidney, with a deficiency in Bcl-2 producing such malformation that renal failure and death result. As a corollary, its role in renal disease states in the adult has been sought. Ischaemia is one of the most common causes of both acute and chronic renal failure. The section of the kidney that is most susceptible to ischaemic damage is the outer zone of the outer medulla. Within this zone the proximal tubules are most sensitive and often die by necrosis or desquamate. In the distal nephron, apoptosis is the more common form of cell death. Recent results from our laboratory have indicated that ischaemia-induced acute renal failure is associated with up-regulation of two anti-apoptotic Bcl-2 proteins (Bcl-2 and Bcl-XL) in the damaged distal tubule and occasional up regulation of Bax in the proximal tubule. The distal tubule is a known reservoir for several growth factors important to renal growth and repair, such as insulin like growth factor-1 (IGF-1) and epidermal growth factor (EGF). One of the likely possibilities for the anti-cell death action of the Bcl-2 genes is that the protected distal cells may be able to produce growth factors that have a further reparative or protective role via an autocrine mechanism in the distal segment and a paracrine mechanism in the proximal cells. Both EGF and IGF-1 are also up regulated in the surviving distal tubules and are detected in the surviving proximal tubules, where these growth factors are not usually synthesized. As a result, we have been using in vitro methods to test: (i) the relative sensitivities of renal distal and proximal epithelial cell populations to injury caused by mechanisms known to act in ischaemia-reperfusion; (ii) whether a Bcl-2 anti-apoptotic mechanism acts in these cells; and (iii) whether an autocrine and/or paracrine growth factor mechanism is initiated. The following review discusses the background to these studies as well as some of our preliminary results. PMID- 10361262 TI - Development of a chronic chromoblastomycosis model in immunocompetent mice. AB - An experimental model to study chromoblastomycosis caused by Fonsecaea pedrosoi was developed in immunocompetent BALB/c mice. Unlike previous models of chromoblastomycosis, in this model a chronic, progressive infection mimicking the infection in humans was developed. This model may be suitable for use in experimental studies of chromoblastomycosis. PMID- 10361263 TI - Cloning and use of the WdURA5 gene as a hisG cassette selection marker for potentially disrupting multiple genes in Wangiella dermatitidis. AB - A genomic clone encoding the Wangiella dermatitidis orotidine monophosphate pyrophosphorylase gene (WdURA5) was isolated by screening a subgenomic plasmid DNA library of this phaeohyphomycotic agent using a PCR amplification product of the gene as a probe. When plasmid DNA containing the cloned WdURA5 gene was introduced by electroporation into a wdura5 auxotrophic recipient strain derived previously by selection with 5-fluoroorotic acid (5-FOA), an apparent gene repair event occurred at high frequency without any evidence of integration of the plasmid DNA. Therefore, the hygromycin B resistance gene (the hph gene) was used as a dominant selective marker for the disruption of WdURA5 to generate a new, more stable, wdura5 auxotrophic strain. Transformation of this strain was then achieved with high efficiency and high frequency by site-specific integration using WdURA5 as a selective marker. To initiate attempts to use this marker repeatedly for multiple chitin synthase (WdCHS) gene disruptions in single strains of W. dermatitidis, a hisG_WdURA5_hisG cassette was constructed and used to disrupt WdCHS2. The WdURA5 gene in the disruptant was then successfully recycled under selection for resistance to 5-FOA. PMID- 10361264 TI - A randomized, comparative trial of treatment of kerion celsi with griseofulvin plus oral prednisolone vs. griseofulvin alone. AB - Glucocorticoids are often recommended along with oral antifungals in the treatment of kerion celsi. In this randomized study, the efficacy of combination therapy with oral griseofulvin and oral prednisolone (n =17) was compared to oral griseofulvin alone (n=13) in the treatment of kerion celsi. Both groups were treated with oral griseofulvin for 8 weeks whereas oral prednisolone was given in tapering doses for 3-4 weeks to the first group only. The final evaluation at week 12 showed a cure rate of 100% in both groups without any significant difference in terms of clinical or mycological cure (P>0.05). No adverse events were noted in either group. In our opinion the combination of oral prednisolone with griseofulvin does not result in additional objective or subjective improvement compared to griseofulvin alone in cases with kerion celsi. PMID- 10361265 TI - Rapid identification of Debaryomyces hansenii/Candida famata by polymerase chain reaction. AB - Primers designed in this study were used in a polymerase chain reaction test to amplify a species-specific fragment of approximately 340 bp of the large subunit ribosomal DNA of Debaryomyces hansenii/Candida famata. None of the other medically relevant yeasts including C. guilliermondii, and also the related species, D. nepalensis and C. saitoana, were amplified by this primer pair. PMID- 10361266 TI - Phylogeny and taxonomy of the family Arthrodermataceae (dermatophytes) using sequence analysis of the ribosomal ITS region. AB - The internal transcribed spacer (ITS) region, covering the ITS1, ITS2 and 5.8S ribosomal DNA was used to evaluate phylogenetic relationships within the fungal family Arthrodermataceae. Sequences of variable length, ranging between 522 and 684 base pairs were aligned. An unrooted consensus tree based on parsimony analysis showed Trichophyton to be polyphyletic, and Microsporum to be paraphyletic. Non-monophyly of these two genera is in conflict with traditional classification. But this relation is not strongly supported by bootstrap analysis. Phylogenetic analysis showed that the two known members of the genus Epidermophyton grouped widely apart from each other. Within Trichophyton, our results suggest a separation of human pathogenic species and primarily geophilic species. Bootstrap support for these two groups is fairly high and both groups are recognized by current taxonomy. Three lineages were revealed within the T. mentagrophytes species complex. Microsporum canis, M. audouinii and M. equinum were found to be closely related. The topology of the tree was robust to various methods of analysis (parsimony and distance) and a different weighting scheme. Weighting of transversions over transitions did not improve the status of poorly supported branches of the tree. PMID- 10361267 TI - Identification, N-terminal region sequencing and similarity analysis of differentially expressed proteins in Paracoccidioides brasiliensis. AB - Paracoccidioides brasiliensis is the causal agent of paracoccidioidomycosis, which is a systemic mycosis in Latin America. This human pathogen is a dimorphic fungus existing as mycelium (26 degrees C) and in infected tissues as a yeast form (36 degrees C). The in vitro differentiation process is reversible and dependent on temperature shift. In the present study, the total proteins from both forms of P. brasiliensis (isolate Pb01) were analysed by two-dimensional electrophoresis. Differentially expressed proteins were identified. Two of these proteins, PbM46 (mycelium) and PbY20 (yeast), were submitted to automated protein sequencing of their N-terminal regions. The 15 amino acid residue sequence of PbM46, AITKIFALKVYDSSG, is similar to enolases from several sources, and specially those from Saccharomyces cerevisiae (80%) and Candida albicans (67%), when compared to the NR database at NCBI using the BLASTP program. The 34 amino acid residue sequence of PbY20, APKIAIVFYSLYGHIQKLAEAQKKGIEAAGGTAD, could probably represent an allergen protein since it is very similar (90%) to the minor allergen protein of Alternaria alternata and 82% similar to the allergen protein of Cladosporium herbarum. This comparative analysis of proteins from mycelium and yeast forms has allowed the identification and characterization of differentially expressed proteins, probably related to differential gene expression in P. brasiliensis. PMID- 10361269 TI - Exserohilum rostratum causing keratitis in India. AB - A case of mycotic keratitis due to atypical Exserohilum rostratum is reported in a 42-year-old male with Hansens disease. PMID- 10361268 TI - Humoral and cellular immune response to a crude exo-antigen and purified keratinase of Microsporum canis in experimentally infected guinea pigs. AB - In order to understand better the host-parasite relationship and to compare with previous observations in Microsporum canis naturally infected cats, the humoral and cellular immune responses to both a crude exo-antigen and a 31.5 kDa purified keratinase were evaluated in 12 M. canis experimentally infected guinea pigs. Humoral and cellular responses were assessed by ELISA from days 0 to 56 postinfection (PI) and by measurement of delayed-type hypersensitivity (DTH) responses on days 14 and 57 PI, respectively. Additionally, immunohistochemical staining was performed and demonstrated that the keratinase was produced in infected guinea pig skin, as previously reported in cats. Despite a marked interindividual variation, all the guinea pigs produced specific IgG to the crude exo-antigen from day 21 PI onwards, but no anti-keratinase IgG was detected. Strongly positive DTH responses to the exo-antigen were observed on both dates, whereas the keratinase elicited no and weak DTH on days 14 and 57 PI, respectively. These results are in agreement with those previously described for naturally infected cats, and indicate that the 31.5 kDa keratinase is not a major antigen in M. canis infection. PMID- 10361270 TI - Microsphaeropsis olivacea as an etiological agent of human skin infection. AB - Microsphaeropsis olivacea is reported as the agent of a case of human skin infection in an otherwise healthy woman. This fungus has not been reported previously as causing disease in humans. It was identified on the basis of its production of pycnidia and conidial structures typical of the Coelomycetes, and by its light brown, ellipsoid to cylindrical and thick walled conidia. The in vitro inhibitory activity of amphotericin B, fluconazole, flucytosine, itraconazole, ketoconazole and miconazole was determined. PMID- 10361271 TI - Ultrastructural characterization of the agent of systemic yeast infection of owl monkeys. AB - Systemic infection by an unclassified yeast-like organism has been encountered sporadically in wild-caught owl monkeys (Aotus sp.) from South America. The infection is presumably acquired in the wild; the incubation period ranges from months to years. The disease is indolent and clinical signs are non-specific. The diagnosis is based on histopathologic observation of yeast-like cells in multiple internal organs. Most cells appear to be phagocytized by macrophages, however, many are apparently free in the extracellular space. Other inflammatory infiltrates, including neutrophils, lymphocytes, plasma cells, and multinucleated giant cells, are conspicuously absent. Cells are thick-walled, globose to oval, range from 5 to 8 micron in diameter, and reproduce by narrow-based budding of single daughter cells. Attempts to cultivate the organism on artificial media have failed. Yeast cell ultrastructure was studied using transmission electron microscopy. The cell wall is multilayered, and the internal structure is markedly heterogenous. In some cells, the cytoplasm is lightly electron-opaque, finely granular and lacks recognizable organelles or nuclei. In others, the cytoplasm is electron dense and contains mitochondria, ribosome-like granules, and a multilobulated nucleus. This organism differs from other recognized pathogenic yeast in its combined light microscopic appearance, organ involvement and host response. Ultrastructurally, it most closely resembles Loboa loboi. PMID- 10361272 TI - The protein kinase C-mediated MAP kinase pathway involved in the maintenance of cellular integrity in Saccharomyces cerevisiae. AB - Signal transduction mediated by the single yeast isozyme of protein kinase C (Pkc1p) is essential for the maintenance of cellular integrity in this model eukaryote. The past few years have seen a dramatic increase in our knowledge of the upstream regulatory factors that modulate Pkc1p activity (e.g. Tor2p, Rom1p, Rom2p, Rho1p, Slg1p, Mid2p) and of the downstream targets of the MAP kinase cascade triggered by it (e.g. Rlm1p, SBF complex). The picture that has emerged connects this pathway to a variety of other cellular processes, such as cell cycle progression (Cdc28p, Swi4p), mating (Ste20p), nutrient sensing (Ira1p), calcium homeostasis (calcineurin, Mid2p, Fks2p) and the structural dynamics of the cytoskeleton (Spa1p, Bni1p). PMID- 10361273 TI - Introns are necessary for mRNA accumulation in Schizophyllum commune. AB - The cDNA coding sequence of the Agaricus bisporus hydrophobin gene ABH1 under the regulation sequences of the Schizophyllum commune SC3 hydrophobin gene gave no expression in S. commune. In contrast, the genomic coding sequence (containing three introns) produced high levels of ABH1 mRNA when transformed to S. commune in the same configuration. Apparently, introns were needed for the accumulation of mRNAs from the ABH1 gene. When the effect of intron deletion on expression of the homologous genes SC3 and SC6 was examined, it was observed that only the genomic coding sequences were expressed in S. commune. Run-on analysis with nuclei harbouring intron-containing and intronless SC6 showed that this effect did not occur at the level of transcription initiation: genomic and cDNA sequences were equally active in this respect. When a 50 bp artificial intron containing the consensus splice and branch sites of S. commune introns, in addition to random-generated sequences, was introduced in the right orientation into the intronless SC3 transcriptional unit, accumulation of SC3 mRNA was restored. By polymerase chain reaction amplification, no unspliced SC3 mRNA species could be detected. Furthermore, the addition of an intron into the transcriptional unit of the gene for green fluorescent protein (GFP) effected clear fluorescence of the transgenic hyphae. Apparently, splicing is required for the normal processing of primary transcripts in S. commune. PMID- 10361274 TI - Bacterial secreted proteins are required for the internaliztion of Campylobacter jejuni into cultured mammalian cells. AB - Presented here is the first evidence that Campylobacter jejuni secrete proteins upon co-cultivation with host cells and in INT 407 cell-conditioned medium. A C. jejuni gene designated ciaB for Campylobacter invasion antigen B was identified, using a differential screening technique, which is required for this secretion process and the efficient entry of this bacterium into a host cell. The C. jejuni ciaB gene encodes a protein of 610 amino acids with a calculated molecular mass of 73 154 Da. The deduced amino acid sequence of the CiaB protein shares similarity with type III secreted proteins associated with the invasion of host cells from other more extensively characterized bacterial pathogens. In vitro binding and internalization assays revealed that the binding of C. jejuni ciaB null mutants was indistinguishable from that of the parental isolate, whereas a significant reduction was noted in internalization. Confocal microscopic examination of C. jejuni-infected cells revealed that CiaB was translocated into the cytoplasm of the host cells. Culturing C. jejuni with INT 407 cells or in INT 407-conditioned medium resulted in the secretion of at least eight proteins, ranging in size from 12.8 to 108 kDa, into the culture medium. C. jejuni ciaB null mutants were deficient in the secretion of all eight proteins, indicating that CiaB is required for the secretion process. The identification of the C. jejuni ciaB gene represents a significant advance in understanding the molecular mechanism of C. jejuni internalization and the pathogenesis of C. jejuni-mediated enteritis. PMID- 10361275 TI - Characterization of a sugar-binding protein from Azospirillum brasilense mediating chemotaxis to and uptake of sugars. AB - Our approach to the isolation of plant-inducible bacterial genes of Azospirillum brasilense, based on the analysis of protein patterns of bacteria grown in the presence and in the absence of plant root exudates, led to the identification of an acidic 40 kDa protein. Cloning and sequencing analysis of the corresponding coding DNA region revealed the presence of two open reading frames transcribed in the same orientation. The deduced ORF1 protein, which corresponds to the 40 kDa protein, is very similar to the periplasmic ChvE protein, identified in Agrobacterium tumefaciens and involved in enhanced virulence. The deduced ORF2 protein shows homology to members of the LysR family of transcriptional regulators. The function of the ChvE-like protein in A. brasilense was investigated further. The protein, designated as SbpA (sugar binding protein A), is involved in the uptake of D-galactose and functions in the chemotaxis of A. brasilense towards several sugars, including D-galactose, L-arabinose and D fucose. Expression of the sbpA gene requires the presence of the same sugars in the growth medium and is enhanced further in combination with carbon starvation of A. brasilense cells. PMID- 10361276 TI - Characterization of the dual start motif of a class II holin gene. AB - Holins are small membrane proteins that, at a genetically programmed time in a bacteriophage infective cycle, allow bacteriolytic enzymes, or endolysins, to escape to the periplasm and to attack the cell wall. Most holins fall into two sequence classes, I and II, based on the number of potential transmembrane domains (three for class I and two for class II). The prototype class I holin gene, S lambda, has a dual start motif and encodes not only the effector holin, Slambda105, but also an inhibitor, Slambda107, with a Met-Lys ...extension at the terminus. The prototype class II holin gene of phage 21, S 21, begins with the motif Met-Lys-Ser-Met ..., and a potential RNA secondary structure overlaps the Shine-Dalgarno sequence. Here, we demonstrate that (i) two protein products are elaborated from S 21, S2171 and S2168; (ii) the shorter product is required for lysis; (iii) the longer product, S2171, inhibits S 21 function; and (iv) the Lys 2 residue is important for the inhibitor function. Moreover, the RNA stem-loop structure is involved in the downregulation of S2171 synthesis. However, our results suggest that, in S 21, different segments of the single consensus Shine Dalgarno sequence serve the two translational starts. These results show that the dual start motifs of class II holin genes are functionally homologous to those of class I holin genes. PMID- 10361277 TI - Heat-induced cell cycle arrest of Saccharomyces cerevisiae: involvement of the RAD6/UBC2 and WSC2 genes in its reversal. AB - The Saccharomyces cerevisiae RAD6 (UBC2 ) gene encodes a ubiquitin-conjugating enzyme that is involved in a wide range of cellular processes including DNA repair, sporulation and N-end rule protein degradation. Under mild heat stress conditions (37-38 degrees C) rad6 null and rad6-149 mutant cells are unable to grow. The molecular basis for this failure to grow is unknown. Here we show that the heat sensitivity of rad6 mutants is not due to cell death but to an inability to progress in the cell cycle. The temperature-induced cell cycle arrest of these mutants is due to a block in a branch of the RAD6 pathway distinct from the DNA repair and the N-end rule protein degradation pathways. Wild-type cells heated to 38 degrees C arrest transiently in the late G1 phase and then resume growth. At 38 degrees C rad6 mutant cells arrest in late G1 but, unlike wild-type cells, are unable to resume cell cycle progression. In both wild-type and in rad6 mutant cells, CLN1 and CLN2 transcript levels fall sharply upon temperature increase. In wild-type cells levels of these transcripts recover rapidly, whereas in the rad6 mutant they recover slowly. As rad6 cells remain arrested even after CLN1 and CLN2 mRNAs regain their preheat stress levels, factors additional to reduced G1 cyclin gene expression must cause the temperature-induced cell cycle block of the mutant. To identify genes involved in the relief of the cell cycle arrest under heat stress, we screened a multicopy yeast genomic library for clones that restore the growth of the rad6-149 mutant. A plasmid was isolated carrying the WSC2 gene, which is closely related to WSC1/SLG1/HCS77, a putative membrane heat sensor. Overexpression of WSC2 reverses the heat-induced cell cycle arrest of rad6-149 but not of rad6 null mutants. Taken together the findings point to the existence of an unidentified heat stress-activated cell cycle checkpoint pathway, which is antagonized by Rad6p by a mechanism also involving Wsc2p. PMID- 10361278 TI - The product of the SNF2/SWI2 paralogue INO80 of Saccharomyces cerevisiae required for efficient expression of various yeast structural genes is part of a high molecular-weight protein complex. AB - Structural genes of phospholipid biosynthesis in the yeast Saccharomyces cerevisiae are activated by the Ino2p/Ino4p transcription factor that binds to ICRE promoter motifs and mediates maximal gene expression in the absence of inositol. We identified the ino80 mutation causing inositol auxotrophy as a result of a defect in ICRE-dependent gene activation. The product of the corresponding wild-type gene INO80 (= YGL150C) shows significant similarity to the Snf2p family of DNA-dependent ATPases. Nevertheless, SNF2 in increased gene dosage did not suppress ino80 mutant phenotypes. Mutation of the Ino80p lysine residue corresponding to the NTP binding site of Snf2p led to a non-functional protein. In ino80 null mutants, gene activation mediated by an ICRE decreased to 16% of the wild-type level. Maximal expression of PHO5, GAL1, CYC1 and ICL1 was also significantly reduced. Thus, Ino80p affects several transcription factors involved in unrelated pathways. As demonstrated by gel filtration, Ino80p is part of a high-molecular-weight complex of more than 1 MDa. Similar to what was found for Snf2p, the Ino80p-containing complex may influence the transcriptional level of several unrelated structural genes by functioning as an ATPase that possibly acts on chromatin. PMID- 10361279 TI - In budding yeast, reactive oxygen species induce both RAS-dependent and RAS independent cell cycle-specific arrest. AB - The role of mild oxidative stresses elicited by diethylmaleate (DEM)-induced glutathione depletion in the progression of the yeast cell cycle has been investigated. We found that different wild-type strains are sensitive to oxidative stresses induced by similar DEM doses: approximately 1 mM on YPD plates, 5-10 mM in shaken flasks. At lower doses, DEM caused a transient decrease in growth rate, largely because of a decreased G1-to-S transition. Treatment with higher DEM doses leads to complete growth arrest, with most cells found in the unbudded G1 phase of the cell cycle. DEM treatment resulted in transcriptional induction of stress-responsive element (STRE)-controlled genes and was relieved by treatment with the antioxidant N-acetyl cysteine. Reciprocal shift experiments with cdc25 and cdc28 mutants showed that the major cell cycle arrest point was located in the Start area, at or near the CDC25-mediated step, before the step mediated by the CDC28 cyclin-dependent kinase. The DEM-induced G1 arrest requires a properly regulated RAS pathway and can be bypassed by overexpressing the G1 specific cyclin CLN2. However, cells with either a deregulated RAS pathway or overexpressing CLN2 failed to grow and arrested as budded cells, indicating that a second DEM-sensitive cell cycle step exists. PMID- 10361280 TI - Poly(A) polymerase I of Escherichia coli: characterization of the catalytic domain, an RNA binding site and regions for the interaction with proteins involved in mRNA degradation. AB - Poly(A) polymerase I (PAP I) of Escherichia coli is a member of the nucleotidyltransferase (Ntr) superfamily that includes the eukaryotic PAPs and all the known tRNA CCA-adding enzymes. Five highly conserved aspartic acids in the putative catalytic site of PAP I were changed to either alanine or proline, demonstrating their importance for polymerase activity. A glycine that is absolutely conserved in all Ntrs was also changed yielding a novel mutant protein in which ATP was wastefully hydrolysed in a primer-independent reaction. This is the first work to characterize the catalytic site of a eubacterial PAP and, despite the conservation of certain sequences, we predict that the overall architecture of the eukaryotic and eubacterial active sites is likely to be different. Binding sites for RNase E, a component of the RNA degradosome, and RNA were mapped by North-western and Far-western blotting using truncated forms of PAP I. Additional protein-protein interactions were detected between PAP I and CsdA, RhlE and SrmB, suggesting an unexpected connection between PAP I and these E. coli DEAD box RNA helicases. These results show that the functional organization of PAP I is similar to the eukaryotic PAPs with an N-terminal catalytic domain, a C-terminal RNA binding domain and sites for the interaction with other protein factors. PMID- 10361281 TI - The role of the trehalose system in regulating the maltose regulon of Escherichia coli. AB - The maltose regulon consists of 10 genes encoding an ABC transporter for maltose and maltodextrins as well as enzymes necessary for their degradation. MalK, the energy-transducing subunit of the transport system, acts phenotypically as a repressor of MalT, the transcriptional activator of the mal genes. Using MacConkey maltose indicator plates we isolated an insertion mutation that strongly reduced the repressing effect of overproduced MalK. The insertion had occurred in treR encoding the repressor of the trehalose system. The loss of TreR function led to derepression of treB encoding an enzymeIITre of the PTS for trehalose and of treC encoding TreC, the cytoplasmic trehalose-6-phosphate hydrolase. Further analysis revealed that maltose can enter the cell by facilitated diffusion through enzymeIITre, thus causing induction of the maltose system. In addition, derepression of TreC by itself caused induction of the maltose system, and a mutant lacking TreC was reduced in the uninduced level of mal gene expression indicating synthesis of endogenous inducer by TreC. Extracts containing TreC transformed [14C]-maltose into another 14C-labelled compound (preliminarily identified as maltose 1-phosphate) that is likely to be an alternative inducer of the maltose system. PMID- 10361282 TI - The stationary-phase morphogene bolA from Escherichia coli is induced by stress during early stages of growth. AB - The Escherichia coli morphogene bolA causes round morphology when overexpressed. The expression of bolA is mainly regulated by a sigmas-dependent gearbox promoter bolA1p. Such regulation results in increased relative levels of expression at slow growth rates, as seen with those attained at the onset of stationary phase. We demonstrate that bolA1p is also induced during early logarithmic growth in response to several forms of stress, and that this induction can be partially sigmas independent. Sudden carbon starvation results in a 17-fold increase in mRNA levels derived from bolA1p 1 h after stress imposition. Increased osmolarity results in a more than 20-fold increase after the same period. Considerable increases in bolA1p mRNA levels were also detected as a result of heat shock, acidic stress and oxidative stress, which has been shown to inhibit sigmas translation. The orders of magnitude of bolA1p induction in log phase due to sudden starvation, osmotic shock and oxidative stress surpass the levels reached in stationary phase. Under sudden carbon starvation and osmotic shock, the cells changed their morphology, resembling those cells in which bolA is overexpressed in stationary phase. Increased expression and morphological changes due to sudden carbon starvation and osmotic shock still occur when sigmaS is not present in a rpoS- background. The results show that expression of bolA is not confined to stationary phase, but it can also play an important role in general stress response. We propose that bolA1p stress induction overrides the normal regulation imposed by growth rate, which is strictly the result of sigmaS-directed transcription. PMID- 10361283 TI - Mutational analysis of ComS: evidence for the interaction of ComS and MecA in the regulation of competence development in Bacillus subtilis. AB - The development of Bacillus subtilis genetic competence is a highly regulated adaptive response to stationary-phase stress. A key step in competence development is the activation of the transcriptional regulator ComK, which is required for the expression of genes encoding the products that function in DNA uptake. In log-phase cultures, ComK is trapped in a complex composed of MecA and ClpC, in which it is rendered inactive. The comS gene, contained within the srf operon, is induced in response to high culture cell density and nutritional stress. Its product functions to release active ComK from the complex, allowing ComK to stimulate the transcription initiation of its own gene as well as that of the late competence operons. Western analysis showed that ComS accumulates to maximal levels between T3 and T4, mirroring the pattern of competence cell development and late competence gene expression. Experiments to examine the target of ComS activity in vitro showed that ComS binds to MecA. This is further supported by coimmunoprecipitation using anti-MecA antiserum. To clarify the role of ComS in competence regulation, a system for evaluating the effect of comS and mutant derivatives on the expression of comG, one of the late competence operons, was constructed. comS mutations, created by alanine-scanning mutagenesis, that significantly reduced comG-lacZ expression were clustered within two regions, one at the N-terminus and the other at the C-terminus of ComS. ComSI13 --> A and ComSW43 --> A were selected for further analysis as representative mutants for both regions required for ComS activity. We observed that ComSI13 --> A showed significantly reduced affinity for MecA, whereas ComSW43 --> A showed near normal binding affinity for MecA. The results show that binding to MecA is critical for ComS function, but do not rule out the possibility that ComS possesses other activities. PMID- 10361284 TI - P-type ATPase spf1 mutants show a novel resistance mechanism for the killer toxin SMKT. AB - SMKT, a killer toxin produced by the halotolerant yeast Pichia farinosa KK1, consists of alpha and beta subunits with folding remarkably similar to that of the fungal killer toxin KP4, a Ca2+ channel inhibitor. The budding yeast Saccharomyces cerevisiae is sensitive to SMKT. To understand the killing mechanism of SMKT, we isolated SMKT-resistant mutants of S. cerevisiae and characterized them. Five spf mutants (sensitivity to the P. farinosa killer toxin) fell into a single genetic complementation group, designated spf1. The SPF1 gene was cloned by complementation of the mutant phenotype. The SPF1 gene encodes a putative P-type ATPase of 1215 amino acid residues that contains 12 membrane-spanning regions. Gene disruption revealed that the SPF1 gene is not essential for viability but is required for the sensitivity to SMKT. The spf1 disruptant showed some phenotypes characteristic of glycosylation-defective mutants and secreted underglycosylated invertase. Fluorescence-activated cell sorting analysis and indirect immunofluorescence microscopy showed that SMKT interacts with the cell surface of the resistant cells but not with that of sensitive cells, suggesting a novel resistance mechanism for this toxin. The glycosylation-defective phenotype and possible killer-resistant mechanisms are discussed in comparison with the Golgi Ca2+ pump Pmr1p. PMID- 10361285 TI - ZapA, the IgA-degrading metalloprotease of Proteus mirabilis, is a virulence factor expressed specifically in swarmer cells. AB - The IgA-degrading metalloprotease, ZapA, of the urinary tract pathogen Proteus mirabilis is co-ordinately expressed along with other proteins and virulence factors during swarmer cell differentiation. In this communication, we have used zapA to monitor IgA protease expression during the differentiation of vegetative swimmer cells to fully differentiated swarmer cells. Northern blot analysis of wild-type cells and beta-galactosidase measurements using a zapA:lacZ fusion strain indicate that zapA is fully expressed only in differentiated swarmer cells. Moreover, the expression of zapA on nutrient agar medium is co-ordinately regulated in concert with the cycles of cellular differentiation, swarm migration and consolidation that produce the bull's-eye colonies typically associated with P. mirabilis. ZapA activity is not required for swarmer cell differentiation or swarming behaviour, as ZapA- strains produce wild-type colony patterns. ZapA- strains fail to degrade IgA and show decreased survival compared with the wild type cells during infection in a mouse model of ascending urinary tract infection (UTI). These data underscore the importance of the P. mirabilis IgA-degrading metalloprotease in UTI. Analysis of the nucleotide sequences adjacent to zapA reveals four additional genes, zapE, zapB, zapC and zapD, which appear to possess functions required for ZapA activity and IgA proteolysis. Based on homology to other known proteins, these genes encode a second metalloprotease, ZapE, as well as a ZapA-specific ABC transporter system (ZapB, ZapC and ZapD). A model describing the function and interaction of each of these five proteins in the degradation of host IgA during UTI is presented. PMID- 10361286 TI - Bacterial genes induced within the nodule during the Rhizobium-legume symbiosis. AB - During the symbiosis between the bacterium Rhizobium meliloti and plants such as alfalfa, the bacteria elicit the formation of nodules on the roots of host plants. The bacteria infect the nodule, enter the cytoplasm of plant cells and differentiate into a distinct cell type called a bacteroid, which is capable of fixing atmospheric nitrogen. To discover bacterial genes involved in the infection and differentiation stages of symbiosis, we obtained genes expressed at the appropriate time and place in the nodule by identifying promoters that are able to direct expression of the bacA gene, which is required for bacteroid differentiation. We identified 230 fusions that are expressed predominantly in the nodule. Analysis of 23 sequences indicated that only three encode proteins known to be involved in the Rhizobium-legume symbiosis, six encode proteins with homology to proteins not previously associated with symbiosis, and 14 have no significant similarity to proteins of known function. Disruption of a locus that encodes a protein with homology to a cell adhesion molecule led to a defect in the formation of nitrogen-fixing nodules, resulting in an increased number of nitrogen-starved plants. Our isolation of a large number of nodule-expressed genes will help to open the intermediate stages of nodulation to molecular analysis. PMID- 10361287 TI - In vivo and in vitro studies of major surface loop deletion mutants of the Escherichia coli K-12 maltoporin: contribution to maltose and maltooligosaccharide transport and binding. AB - The trimeric protein LamB of Escherichia coli K-12 (maltoporin) specifically facilitates the diffusion of maltose and maltooligosaccharides through the outer membrane. Each monomer consists of an 18-stranded antiparallel beta-barrel with nine surface loops (L1 to L9). The effects on transport and binding of the deletion of some of the surface loops or of combinations of several of them were studied in vivo and in vitro. In vivo, single-, DeltaL4, DeltaL5, DeltaL6, and double-loop deletions, DeltaL4 + DeltaL5 and DeltaL5 + DeltaL6, abolished maltoporin functions, but not the double deletion DeltaL4 + DeltaL6 and the triple deletion DeltaL4 + DeltaL5 + DeltaL6. While deletion of the central variable portion of loop L9 (DeltaL9v) affected maltoporin function only moderately, the combination of DeltaL9v with the double deletion of loops L4 and L6 (triple deletion DeltaL4 + DeltaL6 + DeltaL9v) strongly impaired maltoporin function and resulted in sensitivity to large hydrophilic antibiotics without change in channel size as measured in vitro. In vitro, the carbohydrate-binding properties of the different loop mutants were studied in titration experiments using the asymmetric and symmetric addition of the mutant porins and of the carbohydrates to one or both sides of the lipid bilayer membranes. The deletion of loop L9v alone (LamBDeltaL9v), of two loops L4 and L6 (LamBDeltaL4 + DeltaL6), of three loops L4, L5 and L6 (LamBDeltaL4 + DeltaL5 + DeltaL6) or of L4, L6 and L9v (LamBDeltaL4 + DeltaL6 + DeltaL9v) had relatively little influence on the carbohydrate-binding properties of the mutant channels, and they had approximately similar binding properties for carbohydrate addition to both sides compared with only one side. The deletion of one of the loops L4 (LamBDeltaL4) or L6 (LamBDeltaL6) resulted in an asymmetric carbohydrate binding. The in vivo and in vitro results, together with those of the purification across the starch column, suggest that maltooligosaccharides enter the LamB channel from the cell surface side with the non-reducing end in advance. The absence of some of the loops leads to obstruction of the channel from the outside, which results in a considerable difference in the on-rate of carbohydrate binding from the extracellular side compared with that from the periplasmic side. PMID- 10361288 TI - The expression of the trpD, trpC and trpBA genes of Streptomyces coelicolor A3(2) is regulated by growth rate and growth phase but not by feedback repression. AB - Transformation of tryptophan auxotrophs of Streptomyces coelicolor A3(2) and subsequent analysis have allowed the identification of four tryptophan biosynthetic genes. Subcloning, complementation of trp strains, nucleotide sequencing of 5.1 kb and 1.95 kb of DNA and subsequent homology comparisons identified the trpC, trpB and trpA genes and trpD gene respectively. The arrangement of genes in the trpCBA cluster is unusual in that trpC is separated by a small open reading frame, trpX, from the potentially translationally coupled trpB and trpA genes. Sequence analysis of the trpD gene revealed the presence of a large mRNA loop structure directly upstream of the trpD-coding region. S1 nuclease mapping studies of trpCXBA have revealed two major potential transcription start points, one just upstream of the trpC gene and the other located upstream of the trpX gene. S1 nuclease mapping of the trpD region revealed four fragment end-points. Quantitative S1 nuclease protection assays and a promoterless catechol dioxygenase reporter gene have revealed that the expression of all these genes is growth phase dependent and growth rate dependent, expression being maximal during early exponential phase and dropping off sharply in late exponential phase. This growth phase-dependent and growth rate-dependent regulation is the first reported in streptomycete primary metabolism. PMID- 10361289 TI - Penicillin tolerance in Streptococcus pneumoniae, autolysis and the Psa ATP binding cassette (ABC) manganese permease. PMID- 10361290 TI - Identification of putative chromosomal origins of replication in Archaea. PMID- 10361291 TI - Whole-genome sequence annotation: 'Going wrong with confidence'. PMID- 10361292 TI - The Escherichia coli ABC transporters: an update. PMID- 10361293 TI - The ipdC promoter auxin-responsive element of Azospirillum brasilense, a prokaryotic ancestral form of the plant AuxRE? PMID- 10361294 TI - Evolutionary relationships among photosynthetic prokaryotes (Heliobacterium chlorum, Chloroflexus aurantiacus, cyanobacteria, Chlorobium tepidum and proteobacteria): implications regarding the origin of photosynthesis. AB - The presence of shared conserved insertions or deletions in proteins (referred to as signature sequences) provides a powerful means to deduce the evolutionary relationships among prokaryotic organisms. This approach was used in the present work to deduce the branching orders of various eubacterial taxa consisting of photosynthetic organisms. For this purpose, portions of the Hsp60 and Hsp70 genes, covering known signature sequence regions, were PCR-amplified and sequenced from Heliobacterium chlorum, Chloroflexus aurantiacus and Chlorobium tepidum. This information was integrated with sequence data for several other proteins from numerous species to deduce the branching orders of different photosynthetic taxa. Based on signature sequences that are present in different proteins, it is possible to infer that the various eubacterial phyla evolved from a common ancestor in the following order: low G+C Gram-positive (H. chlorum) --> high G+C Gram-positive --> Deinococcus-Thermus --> green non-sulphur bacteria (Cf. aurantiacus ) --> cyanobacteria --> spirochaetes --> Chlamydia-Cytophaga Aquifex-flavobacteria-green sulphur bacteria (Cb. tepidum) --> proteobacteria (alpha, delta and epsilon) and --> proteobacteria (beta and gamma). The members of the Heliobacteriaceae family that contain a Fe-S type of reaction centre (RC 1) and represent the sole photosynthetic phylum from the Gram-positive or monoderm group of prokaryotes are indicated to be the most ancestral of the photosynthetic lineages. Among the Gram-negative bacteria or diderm prokaryotes, green non-sulphur bacteria such as Cf. aurantiacus, which contains a pheophytin quinone type of reaction centre (RC-2), are indicated to have evolved very early. Thus, the organisms containing either RC-1 or RC-2 existed before the evolution of cyanobacteria, which contain both these reaction centres to carry out oxygenic photosynthesis. The eubacterial divisions consisting of green sulphur bacteria and proteobacteria are indicated to have diverged after cyanobacteria. Some implications of these results concerning the origin of photosynthesis and the earliest prokaryotic fossils are discussed. PMID- 10361295 TI - Genetic organization of sulphur-controlled aryl desulphonation in Pseudomonas putida S-313. AB - Pseudomonas putida S-313 is able to desulphonate a broad range of aromatic sulphonates to provide sulphur for growth by monooxygenolytic cleavage to yield the corresponding phenol. After miniTn5 transposon mutagenesis of this strain, 11 mutants were isolated that were no longer able to utilize benzenesulphonate as a sulphur source. Three of these mutants were defective in the utilization of all aromatic sulphonates tested, but they grew normally with other sulphur sources. These strains contained independent insertions in the novel 4.2 kb asfRABC gene cluster, encoding a putative reductase (AsfA), a ferredoxin (AsfB), a putative periplasmic binding protein (AsfC), which was localized to the periplasm using alkaline phosphatase fusions, and a divergently oriented fourth gene, asfR, that encoded a LysR-type regulator protein. A further mutant was interrupted in the ssu locus, which includes the gene for a putative desulphonative monooxygenase. Transformation of Pseudomonas aeruginosa with the asfRAB genes was sufficient to allow arylsulphonate utilization by this species, which does not normally use these compounds, suggesting that the AsfAB proteins may constitute an arylsulphonate-specific electron transport system that interacts with a less specific oxygenase. Expression of the asfABC genes in P. putida was induced by benzenesulphonate or toluenesulphonate, and it was repressed in the presence of sulphate in the growth medium. AsfR was a negative regulator of asfABC expression, and toluenesulphonate induced expression of these genes indirectly by reducing the expression of the asfR gene. PMID- 10361296 TI - Characterization of the Pseudomonas syringae pv. tomato AvrRpt2 protein: demonstration of secretion and processing during bacterial pathogenesis. AB - Pseudomonas syringae pv. tomato strain DC3000 (Pst DC3000) expressing avrRpt2 is specifically recognized by plant cells expressing RPS2 activity, resulting in localized cell death and plant resistance. Furthermore, transient expression of this bacterial avrRpt2 gene in plant cells results in RPS2-dependent cell death. This indicates that the AvrRpt2 protein is recognized inside RPS2 plant cells and is sufficient for the activation of disease resistance-mediated cell death in planta. We explored the possibility that Pst DC3000 delivers AvrRpt2 protein to plant cells via the hrp (type III) secretion pathway. We now provide direct evidence that mature AvrRpt2 protein is secreted from Pst DC3000 and that secretion is hrp dependent. We also show that AvrRpt2 is N-terminally processed when Arabidopsis thaliana plants are infected with Pst DC3000 expressing avrRpt2. Similar N-terminal processing of AvrRpt2 occurred when avrRpt2 was stably expressed in A. thaliana. No cleavage of AvrRpt2 was detected in bacteria expressing avrRpt2 in culture or in the plant extracellular fluids. The N terminus of AvrRpt2 was not required for RPS2 recognition in planta. However, this region of AvrRpt2 was essential for Pst DC3000-mediated elicitation of RPS2 dependent cell death in A. thaliana leaves. PMID- 10361297 TI - Multiple lysophosphatidic acid acyltransferases in Neisseria meningitidis. AB - Lysophosphatidic acid (LPA) and phosphatidic acid (PA) are critical phospholipid intermediates in the biosynthesis of cell membranes. In Escherichia coli, LPA acyltransferase (1-acyl-sn-glycerol-3-phosphate acyltransferase; EC 2.3.1.51) catalyses the transfer of an acyl chain from either acyl-coenzyme A or acyl-acyl carrier protein onto LPA to produce PA. While E. coli possesses one essential LPA acyltransferase (PlsC), Neisseria meningitidis possesses at least two LPA acyltransferases. This study describes the identification and characterization of nlaB (neisserial LPA acyltransferase B), the second LPA acyltransferase identified in N. meningitidis. The gene was located downstream of the Tn916 insertion in N. meningitidis mutant 469 and differed in nucleotide and predicted amino acid sequence from the previously characterized neisserial LPA acyltransferase homologue nlaA. NlaB has specific LPA acyltransferase activity, as demonstrated by complementation of an E. coli plsC(Ts) mutant in trans, by decreased levels of LPA acyltransferase activity in nlaB mutants and by lack of complementation of E. coli plsB26,X50, a mutant defective in the first acyltransferase step in phospholipid biosynthesis. Meningococcal nlaA mutants accumulated LPA and demonstrated alterations in membrane phospholipid composition, yet retained LPA acyltransferase activity. In contrast, meningococcal nlaB mutants exhibited decreased LPA acyltransferase activity, but did not accumulate LPA or display any other observable membrane changes. We propose that N. meningitidis possesses at least two LPA acyltransferases to provide for the production of a greater diversity of membrane phospholipids. PMID- 10361298 TI - The identification of three biologically relevant globotriaosyl ceramide receptor binding sites on the Verotoxin 1 B subunit. AB - The Verotoxin 1 (VT1) B subunit binds to the glycosphingolipid receptor globotriaosylceramide (Gb3). Receptor-binding specificity is associated with the terminally linked Galalpha(1-4) Galbeta disaccharide sequence of the receptor. Recently, three globotriose (Galalpha[1-4] Galbeta [1-4] Glcbeta) binding sites per B-subunit monomer were identified by crystallography. Two of these sites (sites I and II) are located adjacent to phenylalanine-30. Site I was originally predicted as a potential Gb3 binding site on the basis of sequence conservation, and site II was additionally predicted based on computer modelling and receptor docking. The third (site III) was also identified by crystallography and is located at the N-terminal end of the alpha-helix. To determine the biological significance of sites II and III, and to support our previous findings of the significance of site I, we examined the binding properties and cytotoxicity of VT1 mutants designed to block Gb3 binding at each site selectively. The Scatchard analysis of saturation-binding data for each mutant revealed that only the amino acid substitutions predicted to affect site I (D-17E) or site II (G-62T) caused reductions in the binding affinity and capacity of VT1 for Gb3. Similarly, those mutations at sites I and II also caused significant reductions in both Vero and MRC-5 cell cytotoxicity (by seven and five logs, respectively, for G-62T and by four and two logs, respectively, for D-17E). In contrast, the substitution of alanine for W-34 at site III did not reduce the high-affinity binding of the B subunit, despite causing a fourfold reduction in the receptor-binding capacity. The corresponding mutant W-34A holotoxin had a two-log reduction in cytotoxicity on Vero cells and no statistically significant reduction on MRC-5 cells. We conclude that the high-affinity receptor binding most relevant for cell cytotoxicity occurs at sites I and II. In contrast, site III appears to mediate the recognition of additional Gb3 receptor epitopes but with lower affinity. Our results support the significance of the indole ring of W-34 for binding at this site. PMID- 10361299 TI - The V-antigen of Yersinia is surface exposed before target cell contact and involved in virulence protein translocation. AB - Type III-mediated translocation of Yop effectors is an essential virulence mechanism of pathogenic Yersinia. LcrV is the only protein secreted by the type III secretion system that induces protective immunity. LcrV also plays a significant role in the regulation of Yop expression and secretion. The role of LcrV in the virulence process has, however, remained elusive on account of its pleiotropic effects. Here, we show that anti-LcrV antibodies can block the delivery of Yop effectors into the target cell cytosol. This argues strongly for a critical role of LcrV in the Yop translocation process. Additional evidence supporting this role was obtained by genetic analysis. LcrV was found to be present on the bacterial surface before the establishment of bacteria target cell contact. These findings suggest that LcrV serves an important role in the initiation of the translocation process and provides one possible explanation for the mechanism of LcrV-induced protective immunity. PMID- 10361300 TI - Phase variation of HpuAB and HmbR, two distinct haemoglobin receptors of Neisseria meningitidis DNM2. AB - We have previously described HpuAB, a two-component receptor that mediates binding to haemoglobin (Hb), haemoglobin-haptoglobin (Hb-Hp) and apo-haptoglobin (Hp). In this communication, we constructed non-polar mutations in the hpuA and hpuB loci to examine the individual roles of HpuA and HpuB. Our results indicate that both HpuA and HpuB are required for the acquisition of Fe from Hb and Hb-Hp. We isolated Hb utilization-positive (Hb+) variants of our Hb utilization-negative (Hb-) hpu mutants at a frequency of 10(-3) and demonstrated that the Hb+ phenotype resulted from the expression of a second Hb receptor, HmbR. Expression of HmbR in DNM2 was found to be controlled by translational frameshifting involving a polyguanine (G) tract located within the hmbR locus. The hpuA locus also contains a poly(G) tract, which suggested that meningococci could phase vary each Hb receptor independently by slip-strand mispairing in the poly(G) tracts found in hpuA and hmbR. Thus, we isolated a naturally occurring Hb- variant of DNM2, designated DNM2 Hb-, which did not express either HpuAB or HmbR. Hb+ variants of DNM2Hb- were selected and examined for HpuAB and HmbR expression. In each instance, acquisition of HpuAB or HmbR expression was correlated with phase variation in the poly(G) tract of each Hb receptor. PMID- 10361301 TI - Pore-forming activity is not sufficient for Legionella pneumophila phagosome trafficking and intracellular growth. AB - Bacterial pathogens often subvert eukaryotic cellular processes in order to establish a replicative niche and evade host immunity. Inhibition of phagosome lysosome fusion is a strategy used by several intracellular bacteria that grow within mammalian cells. It was shown recently that Legionella pneumophila possesses a cytolytic activity that results from the insertion of pores in the macrophage membrane upon contact, and that this activity requires the dot/icm gene products, which are necessary for intracellular growth and phagosome trafficking. Other bacteria that inhibit phagosome lysosome fusion, such as Mycobacterium tuberculosis, demonstrate similar cytolytic activities, which suggests that formation of pores in the phagosome membrane may account for the defects observed in phagosome trafficking. In this study, we identify a new class of L. pneumophila mutant that retains the pore-forming activity found in virulent bacteria, but is defective in phagosome lysosome fusion inhibition and intracellular growth. These data indicate that cytolytic activity is not sufficient for L. pneumophila-induced alterations in phagosome trafficking. Rather, the pore may be a vehicle that facilitates delivery of bacterial-derived effector molecules to the host cell cytoplasm. PMID- 10361302 TI - A Saccharomyces cerevisiae G-protein coupled receptor, Gpr1, is specifically required for glucose activation of the cAMP pathway during the transition to growth on glucose. AB - In the yeast Saccharomyces cerevisiae the accumulation of cAMP is controlled by an elaborate pathway. Only two triggers of the Ras adenylate cyclase pathway are known. Intracellular acidification induces a Ras-mediated long-lasting cAMP increase. Addition of glucose to cells grown on a non-fermentable carbon source or to stationary-phase cells triggers a transient burst in the intracellular cAMP level. This glucose-induced cAMP signal is dependent on the G alpha-protein Gpa2. We show that the G-protein coupled receptor (GPCR) Gpr1 interacts with Gpa2 and is required for stimulation of cAMP synthesis by glucose. Gpr1 displays sequence homology to GPCRs of higher organisms. The absence of Gpr1 is rescued by the constitutively activated Gpa2Val-132 allele. In addition, we isolated a mutant allele of GPR1, named fil2, in a screen for mutants deficient in glucose-induced loss of heat resistance, which is consistent with its lack of glucose-induced cAMP activation. Apparently, Gpr1 together with Gpa2 constitute a glucose-sensing system for activation of the cAMP pathway. Deletion of Gpr1 and/or Gpa2 affected cAPK-controlled features (levels of trehalose, glycogen, heat resistance, expression of STRE-controlled genes and ribosomal protein genes) specifically during the transition to growth on glucose. Hence, an alternative glucose-sensing system must signal glucose availability for the Sch9-dependent pathway during growth on glucose. This appears to be the first example of a GPCR system activated by a nutrient in eukaryotic cells. Hence, a subfamily of GPCRs might be involved in nutrient sensing. PMID- 10361303 TI - The mitosome, a novel organelle related to mitochondria in the amitochondrial parasite Entamoeba histolytica. AB - Ultrastructural analysis of Entamoeba histolytica reveals that this intestinal human pathogen lacks recognizable mitochondria, but the presence in its genome of genes encoding proteins of mitochondrial origin suggests the existence of a mitochondrially derived compartment. We have cloned the full-length E. histolytica gene encoding one such protein, chaperonin CPN60, and have characterized its structure and expression. Using an affinity-purified antibody raised against recombinant protein, we have localized native E. histolytica CPN60 to a previously undescribed organelle of putative mitochondrial origin, the mitosome. Most cells contain only one mitosome, as determined by immunofluorescence studies. Entamoeba histolytica CPN60 has an amino-terminal extension reminiscent of known mitochondrial and hydrogenosomal targeting signals. Deletion of the first 15 amino acids of CPN60 leads to an accumulation of the truncated protein in the cytoplasm. However, this mutant phenotype can be reversed by replacement of the deleted amino acids with a mitochondrial targeting signal from Trypanosoma cruzi HSP70. The observed functional conservation between mitochondrial import in trypanosomes and mitosome import in Entamoeba is strong evidence that the E. histolytica organelle housing chaperonin CPN60 represents a mitochondrial remnant. PMID- 10361304 TI - Evidence for a system of general protein glycosylation in Campylobacter jejuni. AB - A genetic locus from Campylobacter jejuni 81-176 (O:23, 36) has been characterized that appears to be involved in glycosylation of multiple proteins, including flagellin. The lipopolysaccharide (LPS) core of Escherichia coli DH5alpha containing some of these genes is modified such that it becomes immunoreactive with O:23 and O:36 antisera and loses reactivity with the lectin wheat germ agglutinin (WGA). Site-specific mutation of one of these genes in the E. coli host causes loss of O:23 and O:36 antibody reactivity and restores reactivity with WGA. However, site-specific mutation of each of the seven genes in 81-176 failed to show any detectable changes in LPS. Multiple proteins from various cellular fractions of each mutant showed altered reactivity by Western blot analyses using O:23 and O:36 antisera. The changes in protein antigenicity could be restored in one of the mutants by the presence of the corresponding wild type allele in trans on a shuttle vector. Flagellin, which is known to be a glycoprotein, was one of the proteins that showed altered reactivity with O:23 and O:36 antiserum in the mutants. Chemical deglycosylation of protein fractions from the 81-176 wild type suggests that the other proteins with altered antigenicity in the mutants are also glycosylated. PMID- 10361305 TI - C-terminal interactions between the XerC and XerD site-specific recombinases. AB - Studies of the site-specific recombinase Cre suggest a key role for interactions between the C-terminus of the protein and a region located about 30 residues from the C-terminus in linking in a cyclical manner the four recombinase monomers present in a recombination complex, and in controlling the catalytic activity of each monomer. By extrapolating the Cre DNA recombinase structure to the related site-specific recombinases XerC and XerD, it is predicted that the extreme C termini of XerC and XerD interact with alpha-helix M in XerD and the equivalent region of XerC respectively. Consequently, XerC and XerD recombinases deleted for C-terminal residues, and mutated XerD proteins containing single amino acid substitutions in alphaM or in the C-terminal residues were analysed. Deletion of C-terminal residues of XerD has no measurable effect on co-operative interactions with XerC in DNA-binding assays to the recombination site dif, whereas deletion of 5 or 10 residues of XerC reduces co-operativity with XerD some 20-fold. Co operative interactions between pairs of truncated proteins during dif DNA binding are reduced 20- to 30-fold. All of the XerD mutants, except one, were catalytically proficient in vitro; nevertheless, many failed to mediate a recombination reaction on supercoiled plasmid in vivo or in vitro, implying that the ability to form a productive recombination complex and/or mediate a controlled recombination reaction is impaired. PMID- 10361306 TI - PlcR is a pleiotropic regulator of extracellular virulence factor gene expression in Bacillus thuringiensis. AB - Members of the Bacillus cereus group (B. anthracis, B. cereus, B. mycoides and B. thuringiensis) are well-known pathogens of mammals (B. anthracis and B. cereus) and insects (B. thuringiensis). The specific diseases they cause depend on their capacity to produce specific virulence factors, such as the lethal toxin of B. anthracis and the Cry toxins of B. thuringiensis. However, these Bacillus spp. also produce a variety of proteins, such as phospholipases C, which are known to act as virulence factors in various pathogenic bacteria. Few genes encoding these virulence factors have been characterized in pathogenic Bacillus spp. and little is known about the regulation of their expression. We had previously reported that in B. thuringiensis expression of the phosphatidylinositol-specific phospholipase C gene is regulated by the transcriptional activator PlcR. Here we report the identification of several extracellular virulence factor genes by the virtue of their PlcR-regulated expression. These PlcR-regulated genes encode degradative enzymes, cell-surface proteins and enterotoxins. The PlcR-regulated genes are widely dispersed on the chromosome and therefore do not constitute a pathogenic island. Analysis of the promoter region of the PlcR-regulated genes revealed the presence of a highly conserved palindromic region (TATGNAN4TNCATA), which is presumably the specific recognition target for PlcR activation. We found that the plcR gene is also present in and probably restricted to all the members of the B. cereus group. However, although the polypeptide encoded by the B. cereus PlcR gene is functionally equivalent to the B. thuringiensis regulator, the polypeptide encoded by the B. anthracis gene is truncated and not active as a transcriptional activator. PlcR is the first example described of a pleiotropic regulator involved in the control of extracellular virulence factor expression in pathogenic Bacillus spp. These results have implications for the taxonomic relationships among members of the B. cereus group, the virulence properties of these bacteria and the safety of B. thuringiensis-based biopesticides. PMID- 10361307 TI - ADP-ribosylation of oncogenic Ras proteins by pseudomonas aeruginosa exoenzyme S in vivo. AB - The exoenzyme S (ExoS)-producing Pseudomonas aeruginosa strain, 388, and corresponding ExoS knock-out strain, 388deltaexoS, were used in a bacterial and mammalian co-culture system as a model for the contact-dependent delivery of ExoS into host cells. Examination of DNA synthesis and Ras ADP-ribosylation in tumour cell lines expressing normal and mutant Ras revealed a decrease in DNA synthesis concomitant with ADP-ribosylation of Ras proteins after exposure to ExoS producing bacteria, but not after exposure to non-ExoS-producing bacteria. Examination of normal H-Ras, K-Ras and N-Ras by two-dimensional electrophoresis after exposure to bacteria revealed differences in the degree of ADP-ribosylation by ExoS, with H-Ras being modified most extensively. ADP-ribosylation of oncogenic forms of Ras was examined in vivo using cancer lines expressing mutant forms of H-, N- or K-Ras. The mutant Ras proteins were modified in a manner qualitatively similar to their normal counterparts. Using Ras/Raf-1 co immunoprecipitation after co-culture, it was found that exposure to ExoS producing bacteria caused a decrease in the amount of Raf-1 associated with EGF activated Ras and oncogenic Ras. The results from this study indicate that ExoS ADP-ribosylates both normal and mutant Ras proteins in vivo and inhibits signalling through Ras. PMID- 10361308 TI - Neurospora crassa ro-10 and ro-11 genes encode novel proteins required for nuclear distribution. AB - Movement and distribution of nuclei in fungi have been shown to be dependent on cytoplasmic microtubules and the microtubule-associated motor cytoplasmic dynein. We have isolated hundreds of Neurospora crassa mutants, known as ropy, that are defective in nuclear distribution. Three of the ro genes, ro-1, ro-3 and ro-4, have been shown to encode subunits of either cytoplasmic dynein or the dynein activator complex, dynactin. In this report, we describe the isolation and initial characterization of two additional ro genes, ro-10 and ro-11. ro-10 and ro-11 are non-essential genes that encode novel 24 kDa and 75 kDa proteins respectively. Both ro-10 and ro-11 mutants retain the ability to generate long cytoplasmic microtubule tracks, suggesting that the nuclear distribution defect is not caused by a gross defect in the microtubule cytoskeleton. RO10, as well as RO4 (actin-related protein ARP1, the most abundant subunit of dynactin), appears to be required for the stability of RO3 (p150Glued), the largest subunit of dynactin. We propose that ro-10 mutants lack proper nuclear distribution, because RO10 is either a subunit of dynactin and required for dynactin activity or essential for assembly of the dynactin complex. ro-11 mutations have no effect on RO1 or RO3 levels and have only a very slight effect on the localization pattern of cytoplasmic dynein and dynactin. The role of RO11 in the movement and distribution of nuclei in N. crassa hyphae remains unknown. PMID- 10361309 TI - Hierarchical gene regulatory systems arising from fortuitous gene associations: controlling quorum sensing by the opine regulon in Agrobacterium. AB - Conjugation of the Agrobacterium Ti plasmid pTiC58 is regulated by a hierarchy involving induction by the opines agrocinopines A and B and a quorum-sensing system. Regulation by the opines is mediated by the repressor AccR, while quorum sensing is effected by the transcriptional activator TraR and its ligand, the acyl-homoserine lactone signal molecule Agrobacterium autoinducer (AAI). These last two elements combine to activate expression of the tra system at high population densities. Sequence analysis indicated that traR is the fourth gene of an operon, which we named arc, that is transcribed divergently from accR. Complementation analysis of mutations in the genes 5' to traR showed that the other members of the arc operon are not required for conjugation. Analysis of lacZ reporter fusions demonstrated that traR expression is regulated directly by AccR. Deletion analysis showed that AccR-regulated expression of traR initiates from a promoter located in the intergenic region between accR and orfA, the first gene of the arc operon. Reverse transcriptase-polymerase chain reaction (RT-PCR) and primer extension analyses indicated that the arc transcript initiates upstream of orfA and proceeds uninterrupted through traR. These results are consistent with a model in which quorum sensing is subordinate to the opine regulon because traR has become associated with an operon controlled by the opine responsive transcriptional regulator. PMID- 10361310 TI - Multiple hok genes on the chromosome of Escherichia coli. AB - The hok/sok locus of plasmid R1 mediates plasmid stabilization by the killing of plasmid-free cells. Many bacterial plasmids carry similar loci. For example, the F plasmid carries two hok homologues, flm and srnB, that mediate plasmid stabilization by this specialized type of programmed cell death. Here, we show that the chromosome of E. coli K-12 codes for five hok homologous loci, all of which specify Hok-like toxins. Three of the loci appear to be inactivated by the insertion elements IS150 or IS186 located close to but not in the toxin-encoding reading frames (i.e. hokA, hokC and hokE), one system is probably inactivated by point mutation (hokB), whereas the fifth system is inactivated by a major genetic rearrangement (hokD). In the ECOR collection of wild-type E. coli strains, we identified hokA and hokC loci without IS elements. A molecular and a genetic analysis show that the hokA and hokC loci specify unstable antisense RNAs and stable toxin-encoding mRNAs that are processed at their 3' ends. An alignment of the mRNA sequences reveals all the regulatory elements known to be required for correct folding and refolding of the plasmid-encoded mRNAs. The conserved elements include fbi that ensure a long-range interaction in the full-length mRNAs, and tac and antisense RNA target stem-loops that are required for translation and rapid antisense RNA binding of the processed mRNAs. Consistently, we find that the chromosome-encoded mRNAs are processed at their 3' ends, resulting in the presumed translationally active mRNAs. Despite the presence of all of the regulatory elements, the chromosome-encoded loci do not mediate plasmid stabilization by killing of plasmid-free cells. The chromosome-encoded mRNAs are poorly translated in vitro, thus yielding an explanation for the lacking phenotype. These observations suggest that the chromosomal hok-like genes may be induced by an as yet unknown signal. PMID- 10361311 TI - A hydrogenosome with pyruvate formate-lyase: anaerobic chytrid fungi use an alternative route for pyruvate catabolism. AB - The chytrid fungi Piromyces sp. E2 and Neocallimastix sp. L2 are obligatory amitochondriate anaerobes that possess hydrogenosomes. Hydrogenosomes are highly specialized organelles engaged in anaerobic carbon metabolism; they generate molecular hydrogen and ATP. Here, we show for the first time that chytrid hydrogenosomes use pyruvate formate-lyase (PFL) and not pyruvate:ferredoxin oxidoreductase (PFO) for pyruvate catabolism, unlike all other hydrogenosomes studied to date. Chytrid PFLs are encoded by a multigene family and are abundantly expressed in Piromyces sp. E2 and Neocallimastix sp. L2. Western blotting after cellular fractionation, proteinase K protection assays and determinations of enzyme activities reveal that PFL is present in the hydrogenosomes of Piromyces sp. E2. The main route of the hydrogenosomal carbon metabolism involves PFL; the formation of equimolar amounts of formate and acetate by isolated hydrogenosomes excludes a significant contribution by PFO. Our data support the assumption that chytrid hydrogenosomes are unique and argue for a polyphyletic origin of these organelles. PMID- 10361312 TI - Vascular surgical society of great britain and ireland: audit of surgical technique during carotid endarterectomy by intraoperative duplex ultrasonography: trainees compared with consultant AB - BACKGROUND: There has been increasing interest in audit of outcome following surgery. A previous study reported a significant difference in clinical outcome between consultant and trainees but there is no information on technical factors. Duplex ultrasonography before wound closure was used to compare clinical and technical outcome for consultant and trainees. METHODS: Patients underwent endarterectomy (89 by consultant, 60 by six trainees). In the consultant group nine vessels were patched and 36 patients underwent shunting compared with seven and 31 respectively for trainees. RESULTS: In the consultant group there were two deaths, one stroke, one transient ischaemic attack (TIA) and two cranial nerve injuries. The neurological event rate was 2 per cent, and overall stroke and death rate 3 per cent. There were ten residual flaps (11 per cent) (three re explored) and eight kinks (9 per cent). The residual stenosis rate was 10 per cent (nine of 89) and following re-exploration this reduced to 7 per cent. In the trainee group there was one death, two strokes, one TIA and one cranial nerve injury. The neurological event rate was 5 per cent, and the overall stroke and death rate 3 per cent. There were nine flaps (15 per cent) (four re-explored) and 13 kinks (22 per cent). The residual stenosis rate was 22 per cent (13 of 60) which reduced to 15 per cent (nine of 60) following re-exploration. There was no significant difference in clinical outcome between consultant and trainees but there was an increased incidence of technical problems among the trainees (t = 2.12, P < 0.05). CONCLUSION: Intraoperative duplex ultrasonography is a valuable method for assessing surgical technique; it gives immediate feedback to the surgeon, enables corrective measures to be taken and may facilitate training. PMID- 10361313 TI - Vascular surgical society of great britain and ireland: carotid angiography is used more selectively in the asymptomatic carotid surgery trial AB - BACKGROUND: The ongoing Asymptomatic Carotid Surgery Trial (ACST) has randomized more than 1900 patients to determine whether carotid endarterectomy prolongs stroke-free survival compared with best medical treatment alone. Previously the Asymptomatic Carotid Atherosclerosis Study demonstrated that preoperative angiography caused stroke or death in 1 per cent of patients, and many centres have now reduced or abandoned this practice. This study determined the changing practice of carotid angiography in the ACST. METHODS: Collaborating surgeons completed questionnaires annually on their use and method of angiography. Information on patients in the ACST who had angiography at randomization was also obtained. RESULTS: In 1993, 77 per cent of responding collaborators always performed preoperative angiography and 23 per cent used angiography selectively. This trend has reversed: by 1997, 26 per cent always used preoperative angiography, 70 per cent of respondents used preoperative angiography selectively and 4 per cent had abandoned angiography (P < 0.001, chi2 for trend). Information on carotid angiography at randomization has to date been obtained on 1141 patients in the ACST. Some 44 per cent (497 of 1141) had carotid angiography at randomization. Surgical patients had angiography more commonly than those in the medical group (49 versus 42 per cent; P < 0.03, chi2 test). Changes in carotid angiography were analysed by year of randomization. In 1993, 61 per cent of patients randomized had carotid angiography compared with 42 per cent in 1996 and 1997 (P < 0.001, chi2 for trend). The use of carotid angiography was not related to degree of stenosis estimated by Doppler ultrasonography. CONCLUSION: In the ongoing ACST, there is increasingly selective use of carotid angiography. Less than half the patients in this study had carotid angiography and the use of angiography is now decreasing. This has important implications for validation of carotid duplex in this trial and in future studies. PMID- 10361314 TI - Vascular surgical society of great britain and ireland: impact of spontaneous embolization on cognitive function AB - BACKGROUND: Although microembolization may not be associated with focal neurological events following carotid endarterectomy (CEA), it might contribute towards cognitive impairment. METHODS: Forty-nine patients undergoing CEA underwent 30 min of preoperative ipsilateral transcranial Doppler (TCD) monitoring and a battery of tests of cognitive function, and the results were compared with those of age-matched normal controls. RESULTS: Seven patients who were spontaneously embolizing had significantly worse preoperative cognitive function scores than 42 patients with no recorded emboli (P = 0.03). Overall, patients undergoing CEA had a significant cognitive deficit compared with age matched normal controls. CONCLUSION: Patients who have evidence of spontaneous preoperative embolization on TCD monitoring have a greater degree of preoperative cognitive impairment. This finding underlines the potential clinical importance of microembolization and suggests that this phenomenon should be investigated further. PMID- 10361315 TI - Vascular surgical society of great britain and ireland: Re-evaluation of criteria for reoperation in carotid endarterectomy using intraoperative duplex imaging AB - BACKGROUND: The aim of this study was to compare data obtained during surgery with data collected at 6 weeks after operation to evaluate appropriate criteria for reoperation. METHODS: One hundred and twenty consecutive patients undergoing carotid endarterectomy had duplex scans at operation and 6 weeks later. Neurological evaluation was also documented. RESULTS: Of 96 patients who had a normal intraoperative duplex scan by standard criteria, 91 had normal scans at 6 weeks. One occluded an internal carotid artery (ICA) at 6 weeks with no symptoms. Four had kinks or high-grade contralateral lesions leading to velocity enhancement but no filling defect. All were asymptomatic. Twenty-four patients had abnormal intraoperative scans. Thirteen patients had visible kinking of the ICA or reperfusion hyperaemia; 12 of these patients had normal 6-week scans and one had a mild residual kink but no symptoms. Eleven patients had visible colour filling defects and significant velocity enhancement. Nine of these were reopened and refashioned. Subsequent duplex imaging was satisfactory in all cases and 6 week scans were normal. One patient had an occluded ICA at operation and developed a dense stroke after operation. Another had residual raised velocities distally which remained at 6 weeks. This patient had no symptoms. CONCLUSION: Intraoperative velocity measurements alone cannot be relied upon as an indication for reoperation. Significant velocity enhancement combined with a visible filling defect appears to represent a satisfactory criterion for reoperation. There were no complications as a result of reoperation. There was no early restenosis in the whole group and there were no neurological sequelae in any patient with a satisfactory scan using the above criteria. PMID- 10361316 TI - Vascular surgical society of great britain and ireland: near-infrared spectroscopic monitoring of patients undergoing carotid endarterectomy under locoregional anaesthesia AB - BACKGROUND: The level of cerebral desaturation, which is associated with a change in level of consciousness during carotid endarterectomy, was measured by near infrared spectroscopy. METHODS: Patients were recruited in two centres over 24 months. Surgery was performed under deep and superficial cervical block using 0.5 per cent bupivacaine, with temazepam as a premedication. Cerebral oxygenation was measured by Critikon 2020 near-infrared spectrophotometers (Johnson and Johnson Medical, Newport, UK). RESULTS: Forty-nine procedures were performed on 45 patients (39 men; age range 52-84 (mean 68) years). Recordings were made from the ipsilateral frontal site in 38 patients, from the ipsilateral temporal site in 23 and bifrontally in eight patients. Monitoring failed in three subjects. Percentage changes in regional cerebral oxygen saturation are detailed below. CONCLUSION: Significantly different levels of cerebral desaturation occur in patients with neurological compromise during carotid endarterectomy compared with those who are unaffected. PMID- 10361317 TI - Vascular surgical society of great britain and ireland: transcranial doppler ultrasonography as a predictor of haemodynamically significant carotid stenosis AB - BACKGROUND: Transcranial Doppler (TCD) ultrasonography can detect evidence of collateral flow across the anterior communicating artery and/or the posterior communicating artery, which occurs when there is significant alteration of 'inflow' to the brain. The aim of the study was to determine the blood flow velocity produced by a carotid stenosis which produces this haemodynamic effect on the cerebral circulation and evokes collateral circulation. METHODS: Forty eight patients with varying degrees of carotid stenosis (10 per cent to occlusion) who underwent both carotid duplex and TCD examination were reviewed. An ATL HDI 3000 ultrasound system was used for the carotid and TCD studies. The carotid examination recorded peak-systolic velocity (PSV) and end-diastolic velocity (EDV) in the carotid systems bilaterally. TCD recorded Doppler spectra from the bilateral middle cerebral, anterior cerebral, posterior cerebral, intracranial vertebral and basilar arteries. Collateral flow was assessed in two ways: 'intracranial crossover' collateral and 'posterior to anterior' collateral. Each internal carotid artery (ICA), together with the ipsilateral hemisphere, was analysed for the presence or absence of collateral flow. Data were expressed as mean(s.e.m.). RESULTS: The PSV of the group with collateral circulation was 472(14) cm s-1 and that of the group without collateral flow was 164(3) cm s-1 (P < 0.0001, Mann-Whitney test). The respective EDVs were 158(13) and 58(7) cm s-1 (P < 0.0001). CONCLUSION: PSVs and EDVs in the ICA, in conjunction with collateral flow measured by TCD, are indicators of a haemodynamically significant carotid lesion, and provide more information than two-dimensional imaging studies. In the future, parameters set by combining carotid duplex and TCD investigations may represent the 'gold standard' for evaluation of cerebral blood flow. PMID- 10361318 TI - Vascular surgical society of great britain and ireland: limb outcome following failed femoropopliteal polytetrafluoroethylene bypass for intermittent claudication AB - BACKGROUND: Femoropopliteal (FP) bypass using polytetrafluoroethylene (PTFE) is still considered by many surgeons to be a reasonable procedure for severe intermittent claudication (IC) without limb-threatening ischaemia. The consequences of FP graft failure were examined. METHODS: Over 8 years, 54 patients had 55 FP grafts (that subsequently occluded) inserted for severe IC (42 PTFE and 13 vein grafts) above (30) or below (25) the knee. There were no operative deaths. During the same interval a total of 191 FP grafts were placed, 100 of which were vein grafts. Patient demography and risk factor analysis was similar for both groups. RESULTS: Nineteen patients required amputation subsequent to a failed graft, all of these following PTFE grafts. Mean time to occlusion was 12.2 (range 0-79) months. For PTFE grafts, the mean(s. d.) ankle index rose from 0.51(0.14) to 0.95(0.15) after operation but fell to 0.25(0.15) after occlusion, confirming a highly significant deterioration from preoperative levels, which was not seen in vein graft occlusions. CONCLUSION: Long-term FP bypass patency rates with vein are superior to those obtained with PTFE. Failed PTFE grafts show a significant deterioration in pressure indices compared with preoperative values. FP grafts for IC carry an intrinsic risk of limb loss which is much greater when vein is not used (P < 0.001). PMID- 10361319 TI - Vascular surgical society of great britain and ireland: superficial femoral angioplasty AB - BACKGROUND: Superficial femoral angioplasty (percutaneous transluminal angioplasty, PTA) is a widely performed therapeutic modality used throughout the UK in the treatment of intermittent claudication. However, there is still concern over its efficacy in the management of atherosclerotic occlusive disease. Long term outcome was examined in patients undergoing PTA for short (less than 10 cm) occlusions or stenoses. METHODS: Data were collected prospectively for 413 patients undergoing femoral angioplasty and entered into a database for long-term outcome analysis. Patients were seen at 3, 6 and 12 months and at yearly intervals. Doppler ultrasonography and clinical assessment were performed in all patients and duplex imaging was carried out in those in whom there was doubt about patency. Finally, surviving patients were simply questioned as to whether they felt the original PTA to have been worthwhile. RESULTS: Mean follow-up time was 7 (range 2-11) years. Excluding an initial technical failure rate of 8 per cent, cumulative primary patency at 1, 2, 3, 4 and 5 years was 64, 55, 36, 21 and 14 per cent respectively. Moreover, clinical assessment revealed improvement of the presenting complaint in only 40, 29, 17, 9 and 7 per cent respectively. CONCLUSION: Superficial femoral PTA does not appear to be effective in the management of intermittent claudication in most patients. PMID- 10361320 TI - Vascular surgical society of great britain and ireland: prospective randomized double-blind placebo-controlled crossover study to assess the effect of sublingual glyceryl trinitrate in patients with intermittent claudication AB - BACKGROUND: The effect of sublingual glyceryl trinitrate (GTN) on the claudication distance (CD) and maximum walking distance (MWD) of patients with intermittent claudication was assessed. METHODS: Inclusion criteria were: history of intermittent claudication; resting ankle : brachial pressure index (ABPI) of 1.00 or less; fall in ABPI of more than 0.1 following exercise; and patient not taking nitrates. In the first study 22 patients (median age 69 (range 60-73) years, 16 men, five diabetic, median resting ABPI 0.57 (range 0.1-0.64)) had their CD and MWD measured on a treadmill set at 3.2 km h-1 and 10 per cent gradient. They were then randomized to either GTN or placebo spray, and the distances were remeasured. The crossover portion of the study was then completed. In the second study 28 patients (median age 68 (range 45-84) years, 20 men, six diabetic, median resting ABPI 0.57 (range 0.13-0.98)) were randomized to either GTN or placebo and walked at their own pace along a flat corridor for 15 min. Following a rest of 15 min, the crossover portion of the study was completed. Statistical analysis was by the Wilcoxon matched pairs signed rank test. RESULTS: CONCLUSION: GTN can increase the MWD by 19 per cent when patients with intermittent claudication are walked on a treadmill and by 9 per cent when walking at their own pace on a flat gradient. PMID- 10361321 TI - Vascular surgical society of great britain and ireland: analysis of cold provocation thermography in the objective diagnosis of the hand-arm vibration syndrome AB - BACKGROUND: The hand-arm vibration syndrome (HAVS) is the commonest prescribed disease in the UK. Presently the diagnosis is subjective and the need for an objective investigation to support the diagnosis has been highlighted. This study analyses the potential of cold provocation thermography (CPT) to fulfil this role. METHODS: CPT was performed in ten controls (five men, five women; median age 35 (range 24-78) years) and 21 patients with HAVS (20 men, one woman; median age 45 (range 29-81) years). With an infrared camera, a precooling (PC) image was taken and then, following hand cooling in water at a temperature of 5 degrees C for 1 min, further rewarming images were taken every minute for 10 min. RESULTS: Patient finger tip temperatures were significantly cooler than control temperatures at all time points (P < 0.01, Student's t test). The following Table shows the sensitivity, specificity and PPV of CPT. CONCLUSION: CPT provides strong objective evidence to support the clinical diagnosis of HAVS. PMID- 10361322 TI - Vascular surgical society of great britain and ireland: inhibition of systemic fibrinolysis is associated with myocardial injury in patients operated on for ruptured abdominal aortic aneurysm AB - BACKGROUND: Previous work has demonstrated that ruptured abdominal aortic aneurysm (AAA) is associated with systemic thrombin generation and inhibition of systemic fibrinolysis. The procoagulant and hypofibrinolytic state associated with ruptured AAA predisposes to microvascular and macrovascular thrombosis and subsequent myocardial injury. The aim of this study was to determine the relationship between haemostatic derangement and biochemical evidence of myocardial injury in patients operated on for ruptured AAA. METHODS: Ten patients undergoing repair of ruptured AAA were studied. Tissue plasminogen activator (tPA) activity, plasminogen activator inhibitor (PAI) activity, prothrombin fragment (PF) 1 + 2, D-dimer and fibrinogen levels were measured before operation, and immediately before and 5 min after aortic clamp release. Plasma levels of cardiac troponin (cTn) I were measured before operation, and 6 and 24 h after aortic clamp release. RESULTS: There was no relationship between tPA activity, PF 1 + 2, D-dimer or fibrinogen and cTn-I levels at any sampling point. There was, however, a significant positive correlation (Spearman rank test) between PAI activity immediately before (median 38.6 (range 13.0-39.4) units ml 1) and 5 min after (37.2 (10.6-39.4) units ml-1) aortic clamp release, and cTn-I at 6 h (median 3.17 (range less than 0.5 to 71.1) ng ml-1) and 24 h (5.55 (range less than 0.5 to 110) ng ml-1) after aortic clamp release. CONCLUSION: These data strongly support the hypothesis that the inhibition of systemic fibrinolysis which occurs in response to ischaemia and reperfusion during ruptured AAA repair contributes to the development of subsequent myocardial injury. PMID- 10361323 TI - Vascular surgical society of great britain and ireland: long-term survival following repair of ruptured abdominal aortic aneurysm in patients over 75 years of age AB - BACKGROUND: Previous studies have indicated that patients over the age of 75 years have an increased in-hospital mortality rate following repair of ruptured abdominal aortic aneurysm (AAA). The long-term survival of this patient group has not been reported previously, even though this information may have a strong influence on the decision to operate. METHODS: Interrogation of a prospective database identified 272 patients aged 75 years or over (median age 78 (range 75 93) years) admitted between 1983 and 1995. Long-term patient survival data and survival curves for an age- and sex-matched population were obtained from the General Register Office through the Information and Statistics Division. RESULTS: Sixty-nine (25 per cent) of the 272 patients did not undergo operation. Eighty eight (43 per cent) of the 203 patients who had surgery died within the same hospital admission. The 1-, 5- and 10-year survival rates for the remaining 115 patients (median age 77 (range 75-85) years) were 88, 59 and 26 per cent respectively, at a median follow-up of 54 (range 1-157) months. The median life expectancy for this patient group was 69 months. These data are comparable to those of an age- and sex-matched population. CONCLUSION: Many patients over the age of 75 years who present with ruptured AAA are considered unfit for surgery. These data demonstrate that, even in this elderly population, survivors of ruptured AAA repair enjoy a near-normal life expectancy. PMID- 10361324 TI - Vascular surgical society of great britain and ireland: perioperative renal function following endovascular repair of abdominal aortic aneurysm with suprarenal and infrarenal stents AB - BACKGROUND: Endovascular repair (EVR) of abdominal aortic aneurysm (AAA) is feasible for selected patients. Placing an uncovered stent across the origins of the renal arteries may improve fixation and seal of the proximal end of the stent graft. However, this has potential for problems (e.g. renal artery stenosis or microembolization). This study aimed to evaluate the short-term effect of a suprarenal stent on the function of the individual kidney. METHODS: In 30 patients undergoing EVR for AAA, renal function was assessed before and after operation by 99mTc-radiolabelled diethylenetriamine penta-acetate radionuclide renography and daily measurement of serum creatinine levels. Eleven patients had infrarenal stent-grafts using an aorta uni-iliac system (group 1); 19 patients had the device with an uncovered suprarenal stent (modified Gianturco Z stent), ten of which were aorta uni-iliac and nine bifurcated systems (group 2). Individual kidney function was expressed as the whole kidney transit time (WKTT). In addition, glomerular filtration rate (GFR) was measured from serial blood samples following renography. RESULTS: [Table: see text] CONCLUSION: No result reached statistical significance. Placing an uncovered stent over the origins of the renal arteries does not appear to impair kidney function in the short term. PMID- 10361325 TI - Vascular surgical society of great britain and ireland: prevention of lumbar artery endoleaks following endovascular abdominal aortic aneurysm repair with the selective use of absorbable gelatin sponge AB - BACKGROUND: The aim of endovascular repair of an abdominal aortic aneurysm is to exclude the aneurysm from the systemic circulation in order to prevent aneurysm expansion and rupture. Lumbar artery (LA) endoleaks have been associated with continued expansion of the aneurysm sac and occur with a reported incidence of approximately 10 per cent. The aim of this study was to reduce the incidence of LA or inferior mesenteric artery (IMA) endoleaks. METHODS: Following deployment of an aortic uni-iliac graft an aneurysmograph was performed by injecting contrast into the aneurysm sac to look for patent aortic side branches. Patients with no visible side branches had an occluder deployed in the contralateral iliac artery. Patients with patent side branches had an absorbable gelatin sponge inserted into the aneurysm sac before occluder deployment. Patients had contrast enhanced spiral computed tomography (CT) during follow-up at 1 week, and 3, 6 and 12 months. RESULTS: Fifty-four patients were studied. Eleven were excluded (two perioperative deaths, seven top endoleaks and two occluder endoleaks). Median follow-up was 2 (range 11-7) months. Of the 17 patients in whom patent vessels were demonstrated on an aneurysmograph, one had a patent IMA only, three had a patent LA and IMA, and the remainder had a patent LA. No patient within the study has had a LA or IMA endoleak identified at follow-up CT. CONCLUSION: An intraoperative aneurysmograph can help select patients for aneurysm sac packing with gelatin sponge and has been successful in the short term in preventing LA and IMA endoleaks. PMID- 10361326 TI - Vascular surgical society of great britain and ireland: endovascular aortic aneurysm repair reduces mortality: a physiological analysis AB - BACKGROUND: Endovascular techniques are gaining acceptance as minimally invasive methods for abdominal aortic aneurysm repair, although there have been few studies that provide direct comparison with conventional techniques. A prospective study that compares morbidity and mortality rates following abdominal aortic aneurysm repair, using a physiological scoring system, is presented. METHODS: Between December 1994 and November 1997, 104 elective open aneurysm repairs and 49 endovascular aneurysm repairs were performed at this institution. These patient cohorts were compared using the Portsmouth predictor equation scoring system, which has been shown to predict outcome in vascular surgery. Prospective data from patient notes were used to obtain physiological and operative severity scores that were analysed to predict expected (E) to observed (O) mortality and morbidity rates. RESULTS: Although physiological variables were equivalent in both groups, conventional surgery had a higher mortality (P < 0.05) and morbidity (P < 0.001) rate than endovascular surgery, which was related to the operative severity scores. CONCLUSION: These data suggest that open aortic aneurysm repair has a higher operative severity than endovascular repair which may be reflected in increased mortality rates. PMID- 10361327 TI - Vascular surgical society of great britain and ireland: autologous transfusion reduces blood transfusion requirements in aortic surgery AB - BACKGROUND: Elective aortic surgery is associated with a blood loss that warrants a routine blood crossmatch of 4-6 units. Autologous transfusion strategies to reduce blood requirements were evaluated in a pilot study involving six hospitals in the North West. METHODS: Eighty patients undergoing elective aortic surgery were randomized to either autologous (a combination of acute normovolaemic haemodilution and intraoperative cell salvage) or homologous transfusion. The transfusion trigger, in the absence of pressing clinical need, was 8 g dl-1 haemoglobin for both groups. RESULTS: Randomization achieved two groups well matched for age, aneurysm or occlusive surgery, aspirin intake, estimated blood volume, preoperative haemoglobin and aneurysm size. In the 'best' hospitals (n = 49) mean(s.d.) blood loss (630(49) ml) was significantly lower (P < 0.01) than that in the 'worst' hospitals (1077(110) ml, n = 31) and fewer patients required transfusion (nine of 42 versus 15 of 30; P < 0.05). No significant differences were found between homologous and autologous groups for all variables measured in the 'best' hospitals. In the 'worst' hospitals blood requirements were significantly higher (P < 0.05) for the homologous group (800(112) ml) compared with the autologous group (489(65) ml), although blood loss was similar (1239(195) versus 915(92) ml respectively). CONCLUSION: Autologous transfusion techniques significantly reduced homologous blood requirements in aortic surgery where blood loss exceeded 800 ml. PMID- 10361328 TI - Vascular surgical society of great britain and ireland: randomized double-blind study of dopexamine versus placebo in aortic surgery AB - BACKGROUND: Mechanisms involved in the development of colon- ic ischaemia are not fully understood and there are conflicting reports regarding predisposing factors. The aim of this study was to evaluate the effect of dopexamine hydrochloride on the incidence of colonic ischaemia following aortic surgery and to correlate immunohistochemical markers of inflammatory activation in its pathogenesis. METHODS: Thirty patients, of mean age 65 (range 46-84) years, undergoing elective infrarenal aortic surgery were randomized to receive a perioperative infusion of either dopexamine 2 &mgr;g kg-1 min-1 (n = 12) or 0.9 per cent saline placebo (n = 18). All patients underwent colonoscopy and biopsy following induction of anaesthesia and at 1 week after operation. Sections were stained with haematoxylin and eosin, and for mast cell tryptase (MCT), myeloperoxidase (MPO) and both the inducible (iNOS) and endothelial (eNOS) isoforms of nitric oxide synthase. Sections were analysed blindly and independently by two histopathologists. Patient and operative data were collected and stored separately. RESULTS: Colonic ischaemia was noted in nine patients based on microscopic findings. Endoscopy alone had a sensitivity of 56 per cent. There was a significantly lower incidence of colonic ischaemia in patients receiving dopexamine compared with placebo (P < 0.05). One death resulted from colonic infarction in the placebo group 11 days after operation. There was increased MPO and MCT expression in patients with histological evidence of ischaemia (P < 0.05); iNOS staining within the vascular (P = 0.001) and lamina propria (P < 0.05) components of the mucosa was also significantly greater. No association was found with eNOS. CONCLUSION: Perioperative dopexamine infusion confers a degree of protection to colonic mucosa following aortic surgery, possibly through an anti-inflammatory effect. PMID- 10361329 TI - Vascular surgical society of great britain and ireland: first year of a fast track carotid duplex service AB - BACKGROUND: The risk of a major stroke after a transient ischaemic attack (TIA) is highest in the first 3 months after the onset of symptoms. Urgent endarterectomy in appropriate cases is recommended through a fast-track one-stop assessment clinic. METHODS: Local general practitioners (GPs) were informed that a duplex scan would be guaranteed within 14 days of referral of any patient who had a recent clearly documented TIA or amaurosis fugax. Referral letters were faxed and the scan was authorized by a consultant surgeon. Those with significant disease were seen in the clinic in preparation for operation. Non-significant results were conveyed by post to the GPs with no further action. RESULTS: In the first 12 months of the service, 90 scans were performed through the fast track. In the same interval 490 non-fast-track scans were done after request by a physician (38 per cent), geriatrician (24 per cent), neurologist (14 per cent), vascular surgeon (11 per cent), ophthalmologist (8 per cent) or others (4 per cent). Thirteen (14 per cent) of 90 patients in the fast-track group had carotid endarterectomy, with a median period between referral and operation of 30 (range 20-45) days and median time between onset of symptoms and surgery of 7 (range 4 58) weeks. Endarterectomy was carried out in 12 (2 per cent) of 490 patients in the routine group with a median duration between referral and operation of 127 (range 64-184) days. CONCLUSION: A fast-track service can significantly reduce the time between referral and operation, and increase the number of endarterectomies. Urgent and appropriate referral from the GPs is vital for the service to work efficiently. PMID- 10361330 TI - Vascular surgical society of great britain and ireland: influence of contralateral stenosis on long-term mortality rate following carotid surgery AB - BACKGROUND: Recent evidence suggests that high-grade contralateral stenosis has an adverse effect on perioperative morbidity in carotid endarterectomy (CEA). The relationship between contralateral high-grade stenosis and long-term survival after CEA was examined. METHODS: Three hundred and twenty-seven patients undergoing 333 CEA operations were entered prospectively into a database and long term follow-up was instituted. Cardiac and stroke risk factors were identified before operation and correlated with long-term survival and cause of death. RESULTS: Mean age at operation was 68 (range 42-86) years. Median follow-up was 2 (range 1-8) years. There were 45 deaths (seven perioperative), 17 myocardial, 16 from stroke (four perioperative), five from neoplasia, three respiratory and four others. The cumulative 5-year survival rate was 75 per cent. Patients with high grade contralateral stenosis (greater than 80 per cent) had a significantly reduced life expectancy after CEA (P < 0. 05). CONCLUSION: Severe contralateral carotid disease is not only an adverse perioperative risk factor but also has negative implications for survival of the patient in the longer term. Following CEA, patients in general have a lower life expectancy than a normal age-matched population. PMID- 10361331 TI - Vascular surgical society of great britain and ireland: systemic effects of exercise in claudicants are associated with neutrophil activation AB - BACKGROUND: Claudication induces potentially harmful systemic effects which may be mediated through free radicals and neutrophil activation. This study examined the impact of these mechanisms on renal tubular function. METHODS: Flow cytometry was used to determine CD11b expression by neutrophils and monocytes, and expression of P selectin (CD62P) by platelets, in 16 patients with intermittent claudication and eight matched controls before and at intervals after exercise. Total antioxidant capacity (TAC) and von Willebrand factor (vWF) were measured at similar intervals. Renal tubular function was assessed before and 60 min after exercise by assay of retinol binding protein : creatinine ratio (RBP : Cr) in urine. All patients and none of the controls had a significant exercise-induced fall in ankle pressure. Fluorescein isothiocyanate-labelled monoclonal antibodies against CD11b and CD62P were added to blood and analysis was performed on the flow cytometer within 72 h. RESULTS: There were no significant differences in monocytes, platelets or vWF between patients and controls over time. There was a significant change in RBP : Cr following exercise in patients (median 4.2 (95 per cent confidence interval 2.4-85.2); P 28 days (n = 9) (p = 0.002). Multivariate analysis was applied to identify possible prognostic factors for local control. OTT (p = 0.02) was the only variable that significantly influenced local control. The incidence of radiation ulcer was 33.3% (7/21). Significant indicators of ulceration were fraction size (>> 10 Gy) and NTD (alpha/beta = 3) (>> 130 Gy) (p < 0.05). These results indicate that prolonged OTT was the major reason for the failure of IOC radiotherapy to control local disease and that the relatively high rate of ulceration was due to large fraction size and high NTD (alpha/beta = 3). PMID- 10361415 TI - [Time-resolved three-dimensional contrast-enhanced MR angiography of the carotid artery]. AB - Contrast-enhanced MR angiography (ceMRA) allows practical carotid arteriography without venous enhancement. However, it requires some intricate preparation such as a test bolus of the contrast agent or determination of the tracking volume even in the automatic triggering Smartprep system. The purpose of this study was to obtain carotid ceMRA without any preparation by means of a repeated multiple ultrashort three-dimensional MRA sequence (e3d56), i.e., time-resolved MRA (trMRA). Twenty-three patients underwent sagittal trMRA using a 1.0-Tesla superconducting unit. Multiple projection angiograms are acquired in three contiguous phases with a time resolution of 6 seconds per slab, including 10 partitions, after a bolus injection of 10 ml of Gd-DTPA followed by 20 ml of saline at 2 ml/sec. In all patients, the signal from the arteries could be separated from that of the veins in at least one phase. Carotid trMRA with 6-sec temporal resolution is a reliable technique for selective arteriography, avoiding the necessity of timing the contrast agent bolus. PMID- 10361416 TI - [Evaluation of two-dimensional thick-slice MR DSA: preliminary study]. AB - We have used two-dimensional contrast-enhanced MR angiography for a single thick slice and called it MR DSA. This technique realizes nearly subsecond scanning per image and is therefore superior to other types of contrast-enhanced MR angiography in temporal resolution. To determine the optimal parameters of this technique, we calculated SSR (the signal of brain to the signal of contrast agent ratio) in various settings. We also obtained MR DSA images of 26 cases. We believe that MR DSA may play a supportive role in conventional MR imaging or angiography, since its high temporal resolution is of value in the evaluation of cerebrovascular diseases. PMID- 10361417 TI - [Effects of chronic exercise on renal function in 5/6 nephrectomized spontaneously hypertensive rats (SHR)]. AB - We assessed the renal effects of chronic treadmill exercise in the remnant kidney model of chronic renal failure. Eight-week-old spontaneously hypertensive rats (SHR) were subjected to 5/6 nephrectomy by removal of the left kidney and infarction of two thirds of the right kidney. We performed two series of experiments. Firstly, we investigated the renal effects of chronic mild treadmill exercise in 5/6 nephrectomized SHR. The SHR were divided into 2 groups: a non exercising group (Non-Ex) and a group conducting mild treadmill running at 20 m/min 0 degree grade for 30 min (Mild-Ex) 5 times/week for 4 weeks. Secondly, we investigated the effects of moderate or severe treadmill exercise in the rats. The SHR were divided into 3 groups: a non-exercising group (Non-Ex), a group conducting moderate treadmill running at 20 m/min 0 degree grade for 60 min (Moderate-Ex) and, a group conducting severe treadmill running at 35 m/min 0 degree grade for 60 min (Severe-Ex) 5 times/week for 4 weeks. Chronic treadmill exercise significantly attenuated the increase in proteinuria and serum total cholesterol levels intensity-dependently. These results were confirmed by morphological analysis of the kidneys. Moderate-Ex provided significantly effective protection against an increase in focal glomerular sclerosis. These results indicate that exercise did not worsen renal function and in contrast, suggest a renal-protective action in this rat model. PMID- 10361418 TI - [The role of nitric oxide in deoxycorticosterone acetate-salt hypertensive rats]. AB - The relationship between nitric oxide (NO) and blood pressure (BP) in deoxycorticosterone acetate-salt hypertensive rats (DOC) was investigated. Although urinary NO2- + NO3- (NOx) excretion (UNOxV) increased 2 weeks after surgery (2W-DOC), UNOxV decreased 4 weeks after surgery (4W-DOC) compared with that of the control. BP and UNOxV did not change in DOC after treatment with L arginine (Arg-DOC). Aorta from 4W-DOC and Arg-DOC had significantly decreased relaxation responses to acetylcholine. Deendothelialized aorta from 4W-DOC and Arg-DOC had significantly decreased relaxation responses to lipopolysaccharide. These data suggest that: 1) transient increase of NO synthesis is accompanied by elevation of BP, but long-term elevation of BP decreases NO synthesis in endothelium and smooth muscle cells; 2) L-arginine supplement has no effect on the development of hypertension nor on NO production by endothelium and smooth muscle cells in DOC. PMID- 10361419 TI - [Clinicopathological and morphometrical analysis of 5 cases from 4 families of fibronectin glomerulopathy]. AB - Fibronectin glomerulopathy (FNG) is an inherited disease, characterized by massive fibronectin (FN) deposits in the glomeruli. We semiquantitatively analyzed glomerular lesions and their progression in 5 cases with FNG from 4 different families: a 4 year-old male, a 19 year-old female, a 27 year-old male, a 58 year-old male (the father of the former case) and a 75 year-old male. All subjects showed a 201 times higher value of mean glomerular-tuft area (GA) and a 2.0 times higher mean number of mesangial cells (No. of MC) relative to control (p < 0.001). Strong positive correlations were observed between GA and the No. of MC (r = 0.86, p < 0.001). The younger cases showed markedly higher value of GA and No. of MC than the older cases. Mean individual capillary luminal area was decreased in all but one case and the mean total capillary luminal area, which is roughly estimated as the glomerular filtration area, was less changed compared with the control. The number of capillary loops tended to increase, indicating elongation of the capillary loops. The fractional area of FN (%FN), collagen IV (%Coll. IV) and laminin (%Lam) were high in all cases except for the first Bx in the 27 year-old case. The %FN strongly correlated with %Coll. IV and %Lam (r = 0.86, r = 0.69, p < 0.001, respectively). Serial biopsy (Bx) with a 10 year interval was examined in the 27 year-old case and his father: GA and No. of MC were increased 1.4 and 1.9 times in the son, compared with his first Bx (p < 0.001, respectively), while no change was observed in the father. The %FN, %Coll. IV and %Lam were significantly increased in their second Bx (p < 0.001). These results suggest that 1) enlargement of the glomeruli in FNG is caused by intraglomerular accumulation of FN, Coll. IV and Lam and proliferated mesangial cells, 2) there is a strong influence from the aging factor, and 3) compensatory elongation of the loops (increase in the capillary luminal area) may maintain the glomerular filtration. PMID- 10361420 TI - [Evaluation of thrombomodulin and tumor necrosis factor-alpha levels in patients with hemolytic uremic syndrome caused by enterohemorrhagic Escherichia coli O157:H7 infection]. AB - To investigate the role of thrombomodulin (TM) and tumor necrosis factor-alpha (TNF-alpha) in hemolytic uremic syndrome (HUS), serum and urinary levels of TM and TNF-alpha were determined in patients with hemorrhagic enterocolitis (HC) of enterohemorrhagic E. coli O157:H7 infection. These patients were divided into two groups: an HUS group consisting of patients with HUS; an HC group consisting of patients without HUS. In the 10 days after the onset of diarrhea, the serum TM and TNF-alpha levels in the HUS group were significantly elevated compared with those in the control group but decreased after 11 days. The serum TM and TNF alpha levels in the HC group were not elevated compared with those in the control group except for one case of TNF-alpha. It is suggested that the serum TM elevation showed the severity of endothelial cell damage. Urinary TM and TNF alpha levels in the HUS group were significantly elevated in the first 10 days. The urinary TNF-alpha levels rapidly decreased after 11 days, while the urinary TM levels were persistently high. From these results, the sustained elevation of the urinary TM levels suggested the persistent presence of renal endothelial cell damage, and the decrement of urinary TNF-alpha levels suggested that TNF-alpha acted as a trigger of the renal endothelial cell damage in the first 10 days. PMID- 10361421 TI - [Assessment of adrenal function on steroid using renal diseases in children using the modified original ACTH tolerance test: comparing nephrotic syndrome with non nephrotic syndrome]. AB - Steroid therapy occupies a very important position in various types of kidney diseases in children. The period of steroid therapy in kidney disease tends to extend over a long time and the amount of medication tends to be high. Inhibition of the adrenal function because of steroid therapy is one of the major side effects requiring considerable care. In the present investigation, we examined the inhibition of adrenal function in various pediatric renal diseases using synthetic ACTH. In this study, we checked the serum 11-OHCS and cortisol levels and found that in nephrotic syndrome, the inhibition already existed at sideration. In other diseases we found inhibition of adrenal function using steroid and improvement of the function on reduction of the steroid dose. Patients who used steroid every other day showed better improvement. Therefore, we suggest that in nephrotic syndrome, the inhibition of adrenal function may participate in sideration in the syndrome. It was also found that reduction of the steroid given every other day inhibited the adrenal function to a lesser extent. We found that our challenge test using ACTH is safe and very useful for determining adrenal function. PMID- 10361422 TI - [Factors related to the QT prolongation in chronic renal failure]. AB - QT prolongation, a risk factor for arrhythmia and cardiac death, is observed in uremic patients. Though hypocalcemia, autonomic nerve dysfunction and cardiac hypertrophy are assumed to cause the uremic QT prolongation, the exact mechanism remains unspecified. We therefore examined factors related to the QT interval in chronic renal failure (CRF). Corrected QT interval (QTc) was significantly prolonged in CRF just before the induction of dialysis therapy (group A) compared with nephrotic syndrome with the intact or mildly impaired renal function (group B). QTc was also prolonged in acute renal failure (group C). Cardio-thoracic ratio, serum albumin and Ca correlated with QTc in group A, but not in B or C. A single HD session in group A failed to shorten QTc, despite a significant increase in serum Ca++. Autonomic dysfunction did not appear to be a major determinant of QT prolongation, since QTc was not different between diabetics and non-diabetics in group A and in chronic HD patients (group D). In group D, QTc did not correlate with SV1 + RV5 on ECG or left ventricular wall thickness (LVWT) on echocardiography. In another group of chronic HD patients (group E), there was no significant correlation between QTc and the parameters of left ventricular mass, plasma brain natriuretic peptide (BNP). However, in the patients subjected to repeated echocardiography in group D, QTc and LVWT changed in parallel. In a retrospective analysis of QTc in group D, QTc was maximally prolonged at the time of starting HD therapy, and gradually improved in the following 1-5 years in both diabetics and non-diabetics. In contrast, chronic CAPD patients (group F) revealed no improvement of QTc. Thus, uremic QT prolongation cannot be explained simply by any of the previously assumed factors, but appears to be affected by multiple factors, which are partially correctable by chronic HD therapy. PMID- 10361423 TI - [A case of nephrotic syndrome associated with myasthenia gravis and malignant thymoma]. AB - A 26-year-old woman who presented facial and lower leg edema associated with massive proteinuria was admitted to our hospital in February 1992. Nine months before this admission, she exhibited myasthenia gravis and malignant thymoma, and underwent total thymectomy. On admission, there was no symptom of myasthenia gravis. She was diagnosed as having nephrotic syndrome and the first renal biopsy was performed. The histological findings showed membranous nephropathy. Immunofluorescent microscopy revealed that IgG and C3 were stained in a granular pattern in the periphery, and subepithelial deposits were observed in the basement membrane of the glomerulus by electron microscopy. With the administration of prednisolone, proteinuria disappeared and the nephrotic syndrome remitted. She was admitted again in January 1993 due to proteinuria and lower leg edema following cystitis. The findings of the second renal biopsy were unremarkable. She was administered cyclosporin A to improve the nephrotic syndrome and to reduce the side effects of prednisolone. The proteinuria disappeared again and this effect was dependent on the dose of cyclosporin A. Since the first administration, no symptoms of myasthenia gravis or malignant thymoma have been observed. The relationships among myasthenia gravis, malignant thymoma and nephrotic syndrome were examined. Although the first renal biopsy findings showed membranous nephropathy, from the therapeutic responses of both prednisolone and cyclosporin A, the main course of proteinuria in this case may have been due to minimal change nephrotic syndrome. We consider this case of nephrotic syndrome to be important considering its etiology and the relationship between the histological findings and its clinical course. PMID- 10361424 TI - [Focal segmental glomerulosclerosis associated with type C virus hepatitis and decrement of proteinuria by interferon-alpha therapy]. AB - Focal segmental glomerulosclerosis (FSGS) associated with type C virus (HCV) hepatitis has not been described in the literature to date. However, we experienced a 30-year-old man, who had had HCV hepatitis, developed nephrotic syndrome and was admitted to our hospital. The first renal biopsy showed FSGS which was diagnosed by light, immunofluorescent, and electron microscopic study. FSGS diagnosis was based upon the findings of focal segmental glomerular sclerosis associated with hyalinosis and foam cells, segmental deposition of IgM and C3 on glomeruli, and epithelial cell vacuolization in the Bowman's space. HCV hepatitis was treated with interferon-alpha (INF-alpha) over 6 months. The treatment brought the disappearance of not only HCV-RNA from the blood, but also the manifestation of nephrotic syndrome. Therefore, the second renal biopsy was performed, but did not reveal any great pathological improvement. Five months later after the remission, he again had an elevated HCV-RNA level and a relapse of nephrotic syndrome. He was retreated with the same therapy and achieved a second remission of nephrotic syndrome. FSGS associated with HCV hepatitis is described first and the implication of INF-therapy in the improvement of proteinuria is discussed. PMID- 10361425 TI - [A case of interstitial nephritis induced by a super antigen produced by methicillin-resistant Staphylococcus aureus (MRSA) presenting as acute renal failure]. AB - We report the case of a 21-year-old man who had been developing acute renal failure with Methicillin-resistant Staphylococcus aureus (MRSA) colitis and sepsis. He was admitted for consciousness disturbance, nausea, vomiting, and diarrhea. Oliguria was also observed and his serum creatinine level was elevated to 10 mg/dl. Urinary protein was positive and an abundance of hyaline cast were seen in urinary sedimentation. Diarrhea and pyrexia were prolonged and serum C reactive proteins were elevated, but lymphocyte and leukocyte counts temporarily decreased from the 3rd to the 6th hospital day and remained low until normalizing after the 14th day. His clinical symptoms improved with hemodialysis (HD) and effective antibiotic therapies. An MRSA strain producing toxic shock syndrome toxin-1 (TSST-1), a super antigen which specifically stimulates human V beta 2 positive T cells, was separated from his feces and blood. To ascertain the cause of his renal dysfunction, a renal biopsy was performed on the 8th day. His renal histology revealed acute interstitial nephritis with severe inflammatory cell infiltration around the medullary areas without glomerular changes. Most of the infiltrated cells were small monocytes, and lymphoid cells were rich in the interstitium. With immunohistochemical staining, over 70% of T-cells were V beta 2-positive. TSST-1-producing MRSA was detected in his blood specimen. Furthermore, V beta 2-positive T cells were accumulated in the renal intersititium, and transient lymphocytopenia was observed. These data suggested the following possible pathogenesis for interstitial nephritis: TSST-1 acts as a super antigen in the renal interstitium where major histocompatibility complex (MHC) is class-2-positive, thereby resulting in interstitial nephritis with T cell migration. PMID- 10361426 TI - [Pathophysiology of the renin-angiotensin system]. AB - Angiotensin II is a potent vasoconstrictor as well as a growth stimulator. Angiotensin type 1 (AT1) and type 2 (AT2) receptors have been cloned, and molecular mechanisms by which angiotensin II stimulates cell growth have been also clarified by molecular biological techniques including gene targeting and transgenic animals. AT1 promotes hypertrophy and hyperplasia through G protein associated pathway and G-protein-independent pathway consisted of tyrosine kinase, JAK/STAT, and growth hormone signal transduction. Function of AT2 is not fully understood, but recently has been reported to reverse the effect of AT1. This is a brief review on angiotensin II functions and its signal transduction, including the aspect of the constitutively active AT1 mutant receptor. PMID- 10361427 TI - [Angiotensin II receptor antagonists: a review of the development and future perspective]. AB - Pharmacological blockade of the renin-angiotensin system has been found to be a safe and efficacious way to treat hypertension and congestive heart failure. The success of the angiotensin converting enzyme inhibitors has led to interest in alternative ways to block the renin-angiotensin system. Angiotensin II receptor antagonists are a new class of anti-hypertensive drugs that provide a specific blockade of the effects of angiotensin II. Losartan potassium, the first compound of this class, has recently been approved in Japan. It seems likely that the angiotensin receptor antagonists will be suitable to first-line therapy and use of this class for treatment of hypertension will dramatically increase in Japan because of the excellent clinical and laboratory safety profiles. PMID- 10361428 TI - [Gene loci and polymorphisms of angiotensin II receptor]. AB - The gene encoding angiotensin II type 1 receptor (AT1) is mapped on 3q21-q25 region, and a polymorphism, A1166C, is located at 3'untranslated region (UTR). A1166C is associated with increased risk for hypertension, aortic stiffness, left ventricular hypertrophy and diabetic nephropathy, and synergistically increases the risk for ischemic heart disease with DD polymorphism of angiotensin converting enzyme gene. However, these results were still in controversy. On the other hand, the gene encoding angiotensin II type 2 receptor (AT2) gene is mapped on Xq22-q23 region, and a polymorphism, C3123A, is identified at 3'UTR of AT2 gene. However, any significant association with C3123A has not been obtained in case control studies yet. PMID- 10361429 TI - [Distribution and function of angiotensin II receptor subtypes--central nervous system]. AB - AT1 receptors are predominant in the brain of monkeys and rabbits, while AT2 receptors are relatively abundant in the rat brain. In the human brain, all of the angiotensin II receptors in the forebrain, midbrain, pons, medulla and spinal cord are AT1 receptors, and AT2 receptors are found only in the cerebellum. Angiotensin II in the brain increases water and sodium intake, raises blood pressure, attenuates baro-reflex function, and increases vasopressin secretion. These cardiovascular actions of angiotensin II are exclusively mediated by AT1 receptors. Since the mice whose AT2 receptors are knocked out show the increase in blood pressure, the decrease in body temperature, and some alterations in behavior, these receptors may also play roles in the central nervous system. PMID- 10361430 TI - [Distribution and function of angiotensin receptor subtypes in cardiovascular system]. AB - Angiotensin II (AII) participates in regulation of arterial blood pressure through its binding to AII receptors distributing among its target organs. In addition, locally produced AII appears to play a major role in the pathogenesis of cardiovascular hypertrophy via mechanism not related to blood pressure. Two subtypes of AII receptors, AT1 and AT2, are recognized as distinct in both molecular and pharmacological basis. In adult, AT1 is a dominant subtype in cardiovascular system, and mediates virtually all the previously known actions of AII, including vasoconstriction, production of growth factors, hypertrophy of smooth muscle and cardiomyocyte, proliferation of smooth muscle and fibroblast, production of extracellular matrix and so on. Recently, upregulation of AT2 expression is revealed under certain pathological conditions, such as vascular injury, myocardial infarction, and heart failure. Biological significance of AT2 are still under investigation, however, countering actions against AT1 are often suggested. PMID- 10361431 TI - [Kidney]. AB - Kidney is one of the major target organ for angiotensin II (AT-II). The presence of three types of AT-II receptors, AT-II type 1, type 2, type 4 receptor (AT1, AT2, and AT4 receptor, respectively) were reported in kidney. AT1 receptor is the main type of receptor in kidney and mediates most of physiological effects of AT II in kidney. The population of AT2 receptor is small in kidney. However, AT2 receptor also seems to play an important role for development and apoptosis in kidney. AT4 receptor is the receptor for angiotensin IV, but the physiological function is still unknown. PMID- 10361432 TI - [Angiotensin II receptor subtype in human adrenal glands]. AB - Although adrenal gland is one of the major target organs of angiotensin II (Ang II), the pathophysiological significance of the its receptor subtype has not been elucidated. We demonstrated by reverse transcription-polymerase chain reaction with Southern blot analysis mRNA expression of both AT1 and AT2 in human adrenal tissues of normal adrenocortical tissues, aldosterone-producing adenoma, Cushing's syndrome, and pheochromocytoma. Ang II-induced aldosterone secretion in vitro was suppressed only by 50% in the presence of selective AT1 antagonist CV 11974, while AT2 agonist CGP-42112 increased aldosterone secretion by 55% over the control. Ang II or CGP-42112 did not affect cortisol secretion. In addition, Ang II could stimulate aldosterone secretion in AT1a knockout mice both in the presence and absence of CV-11974. These results suggest that non-AT1 receptor subtype(s) including AT2, as well as AT1, is involved in the stimulation of aldosterone secretion from human adrenals. PMID- 10361433 TI - [Angiotensin receptor in the lung]. AB - In situ hybridization of angiotensin receptor mRNA and ligand-binding assay showed that main subtype of angiotensin receptor in the lung was type 1(AT1) in pulmonary vessel, whereas type 2(AT2) was not detectable. AT1 induces the pulmonary artery contraction through inositol phosphate-protein kinase C pathway, therefore the non-peptide AT1 antagonist was applied to animal model of pulmonary hypertension (PH). AT1 antagonist improved pulmonary arterial remodeling and right ventricular hypertrophy in rat hypoxia-induced PH but not in rat monocrotaline-induced PH. Less effectiveness of AT1 antagonist for PH might be no AT2 stimualtion under increased angiotensin II level in blood and lung tissue response to AT1 antagonist treatment. PMID- 10361434 TI - [Modulations by stimulation of angiotensin II type 1 receptor of gastrointestinal functions]. AB - Electrolytes and water are bidirectionally transported across the epithelial cells of mammalian small and large intestines. In animals with chronic renal failure, plasma levels of angiotensin II and specific 125I-angiotensin II binding sites increase and result in enhanced excretions of K+, Cl- and urate from distal colon, in an AT1-receptor sensitive manner. Angiotensin-converting enzyme activity and mRNA levels for renin-2 and AT1A-receptor are blunted or lowered transiently by gastric intake of low Na+ in rodent, suggesting that angiotensin II serves as a humoral mediator for "gastric Na+ monitor". Angiotensin II causes also very potent contractions of gastrointestinal smooth muscles as shown in vascular smooth muscles. This review is focused on understanding potential roles of angiotensin II in gastrointestinal functions. PMID- 10361435 TI - [Renin-angiotensin-aldosterone system in the reproductive system]. AB - Besides the circulating renin-angiotensin-aldosterone (R-A-A) system, the tissue R-A-A system has been elucidated to play important roles as autocrines and/or paracrines. The components of R-A-A system are expressed in the ovary, uterus and placenta, indicating the existence of the tissue R-A-A system in these organs. The data indicating the involvement of R-A-A system of these tissues into reproduction have been accumulated. AT2 receptors might modulate the initiation and progression of follicle atresia involving granulosa cell apoptosis. AT2 receptors are expressed abundantly in the uterus and decreased during pregnancy. The placental renin are shown to be secreted into the maternal circulation and elevate blood pressure. It is expected to elucidate the significance of the R-A-A system in the reproductive system. PMID- 10361436 TI - [Structure and function of angiotensin II receptor]. PMID- 10361437 TI - [Signal transduction systems of angiotensin II receptors]. AB - Angiotensin (A) II is a potent constrictor as well as growth stimulant of vascular smooth muscle cell caused by activation of AT1 receptor signal transduction systems. There are two major signal systems of AT1 receptor: one leads to an increase in cytosolic free calcium levels causing smooth muscle contraction which may result in high blood pressure, and the other leads to smooth muscle proliferation and inflammation which may result in atherosclerosis. AT1 receptor activation induces phosphinositide hydrolysis by phospholipase C and creates an inositol phosphate, which release calcium from cytosolic calcium pools. Cytosolic calcium can also be elevated by activation of calcium channel via a link between AT1 receptor and a G protein. Protein phosphorylation triggered by AT1 receptor is important for cell growth, in which tyrosine kinase, serine/threonine kinase and protein kinase C are involved. Free radicals are generated by NADH/NADPH oxidase in response to AT1 receptor activation, causing expression of genes leading to atherosclerosis. On the other hand, activation of AT2 receptor is shown to play a role of lowering blood pressure. Some phosphatases and NO/cyclic GMP would be involved in the mechanism. In renal vasculature, endothelium dependent epoxygenase products are synthesized by AT2 receptor stimulation causing vasorelaxation. In summary, AT1 receptor signals are vasopressive and evoke atherosclerosis, whereas AT2 receptor signals may possibly be vasodilatory. PMID- 10361438 TI - [Role of angiotensin II-forming enzymes, angiotensin-converting enzyme and chymase]. AB - Angiotensin (Ang) II plays a crucial role in regulation of blood pressure and proliferation of vascular tissues. Recent studies have demonstrated that the AngII-forming enzymes, ACE and chymase, are observed in heart and vascular tissues. In isolated human arteries, chymase predominantly converted Ang I to AngII rather than ACE. In hypertensive models, AngII formation by ACE in vascular tissues plays an important role in maintaining hypertension, while that by chymase hardly does. Chymase-dependent AngII formation induces vascular diseases such as neointima formation after balloon injury. AngII receptor antagonists block AngII formation by chymase in addition to ACE and may be useful for cardiovascular diseases rather than ACE inhibitors. PMID- 10361439 TI - [Angiotensin II receptor-mediated function unmasked by gene-engineered animals]. AB - The receptors for angiotensin (Ang) II are classified into two subtypes (AT1-R and AT2-R) by the discovery of non-peptidic ligands and AT1-R mediates most of the cardiovascular actions of Ang II. AT2-R is expressed at very high levels in the developing fetus, whereas in the adult its expression in the cardiovascular system is very low. Cardiac myocyte- or vascular smooth muscle-specific overexpression mice of AT2-R display an inhibitory effect on Ang II-induced chronotropic or pressor actions, suggesting the role of AT2-R on the activity of cardiac pacemaker cells or maintenance of vascular resistance. AT2-R also activates the kinin/nitric oxide/cGMP system in the cardiovascular and renal system, resulting in the AT2-R-mediated cardioprotection, vasodilation and pressure natriuresis. These effects transmitted by AT2-R are mainly exerted by stimulation of protein tyrosine or serine/threonine phosphatases in Gi-protein dependent manner. The expression level of AT2-R is much higher in human hearts than in those of rodents, and the AT2-R-mediated actions are likely enhanced, especially by clinical application of AT1-R antagonists. PMID- 10361440 TI - [The role of angiotensin II in the development of cardiovascular remodeling]. AB - We examined the role of AT1 in the development of cardiovascular remodeling using AT1a knockout (KO) mice. 1. Pressure overload and mechanical stretch induced hypertrophic responses in KO and wild type (WT) cardiomyocytes (CM). Stretch activated MAPK through PKC in WT CM and through tyrosine kinase in KO CM. 2. The number of ventricular premature beats and tachycardia was larger in WT mice than KO mice. 3. Left ventricular remodeling after myocardial infarction was more remarkable in WT mice than KO mice. 4. Vascular injury induced neointimal formation in KO mice as well as in WT mice. PMID- 10361441 TI - [Effects of angiotensin II on the cerebral circulation and function]. AB - The effect of angiotensin II (A II) on cerebral arteries is variable; A II would constrict or dilate cerebral arteries depending on species, the size of vessels examined, preexisting contractile levels, and the presence of functional endothelium. Hence exogenously applied A II may increase or decrease cerebral blood flow. The physiological role played by endogenous A II is even unclear in the cerebral circulation. Endogenous A II is unlikely to modulate cerebral blood flow. However, it may play some roles in the determination of the autoregulation of cerebral blood flow, at least judging from the effect of angiotensin converting enzyme inhibitors. Circulating A II elevates blood pressure, and promotes water and sodium intake. A II derived from the central renin-angiotensin system would play diverse roles as a neurotransmitter or a local hormone. PMID- 10361442 TI - [Angiotensin II and the kidney]. AB - Angiotensin II (AngII) plays a central role for maintenance of GFR and Na balance particularly in volume depletion, when AngII preferentially increases the resistance of efferent arterioles as compared to afferent arterioles, enhancing the glomerular perfusion pressure. In addition, AngII enhances tubular reabsorption of sodium in proximal tubules directly and indirectly as a consequence of glomerulotubular balance. AngII also stimulates Na reabsorption in the collecting ducts by stimulating the release of aldosterone from the adrenal cortex. AngII augments tubuloglomerular feedback. Recently, it has been shown that AngII has a variety of non-hemodynamic effects on cell growth and differentiation as well as inflammatory responses, and it has been speculated that the increased renin-angiotensin, not only systemic but local one, may be responsible for the pathogenesis of a number of renal diseases. PMID- 10361443 TI - [Interaction between angiotensin II and other local vasoactive substances]. AB - Recent advances in the molecular characterization for angiotensin II (A II) related to nitric oxide, endothelin-1, prostaglandin, and adrenomedullin are reviewed. A II, the main biological active peptide of the renin-angiotensin system, plays an important role in cardiovascular homeostasis such as regulation of blood pressure and tissue remodeling. A II produces vasoconstriction by a direct action on smooth muscle cells via AT1 receptor. This action can be due to circulating A II but also to A II produced within the tissues. Nitric oxide and adrenomedullin are potent vasorelaxant substances. These substances may play a role as local antimigration factors, and antagonize the effect of A II. Whereas endothelin-1 and thromboxane A2 are vasoconstrictor substances, and may have a role as growth factors. A II and these vasoactive substances mutually exert several biological actions in cardiovascular diseases. PMID- 10361444 TI - [Angiotensin II and apoptosis]. AB - Angiotensin II (AII), the effector octapeptide of the renin-angiotensin system, exerts a multitude of actions, including vascular contraction, aldosterone secretion, catecholamine release, glycogenolysis, and decreased renal filtration. These diverse actions are mediated through AII receptor subtypes present in a variety of target tissues. Molecular cloning studies have identified two major types of mammalian AII receptors, designated AT1 and AT2, which are classified as a typical family of seven transmembrane guanyl nucleotide-binding protein (G protein) coupled receptors from hydropathy analyses. Mainly, if not all, of the actions of AII are mediated by AT1 that has been well characterized. Recently, AT2 is found to exert growth inhibitory and proapoptotic effects, but its physiological role is still unclear. The molecular mechanism and the physiological importance of signaling pathways via these receptors remain to be elucidated. PMID- 10361445 TI - [Pharmacological properties and its significance in clinical practice]. AB - Losartan is a potent non-peptide, selective angiotensin II (AngII) type 1 (AT1) receptor antagonist. Losartan has been worldwide marketed as the first orally active AT1 receptor antagonist with once-daily dosing for treatment of hypertension. In a study of patients with heart failure, the mortality appeared to be lower with losartan than with the ACE inhibitor captopril. In healthy subjects, losartan produced a dose-dependent reduction in serum uric acid. The mechanism of action is considered to be the inhibition of reabsorption of uric acid in the proximal tubules of the kidney. Furthermore, it was recently reported that losartan has moderate affinity for the thromboxane (TX) A2 receptor in a competitive-inhibition manner in the platelets and vascular smooth muscle. The efficacy of losartan with regard to not only AT1 receptor blockade, but also the reduction of serum uric acid and the blockade of TXA2 receptors, may be advantageous to patients with hypertension having these cardiovascular risk factors. PMID- 10361446 TI - [Pharmacological profiles of candesartan cilexetil (TCV-116)]. AB - Candesartan cilexetil has shown potent and long-lasting antihypertensive effects in clinical trials and in several hypertensive animal models. In rabbit aortic preparation, candesartan, active form of candesartan cilexetil, decreased the maximal contractile response of angiotensin II (insurmountable inhibition). This inhibitory mode was different from that of other angiotensin II receptor blockers, and showed a shift to the right in the angiotensin II-induced contraction curve. In kinetic studies using bovine adrenal cortical membrane and tritiated candesartan, both receptor association and dissociation were found to be slow. The insurmountable inhibition of candesartan in vascular contraction is the result of its tight binding and slow dissociation from AT1 receptors. These characteristics are related to the potency and long duration of action in candesartan cilexetil. PMID- 10361447 TI - [The effects of angiotensin II receptor antagonists on glucose, lipid, and uric acid metabolism in essential hypertensives]. AB - Many of hypertensive individuals have glucose intolerance, dyslipidemia and hyperuricemia. It is important to take care of these metabolic disease for not only the progression hypertension itself but also the prevention of atherosclerosis. We reviewed the effects of angiotensin II receptor antagonists on glucose, lipid, and uric acid metabolism in essential hypertensives. PMID- 10361448 TI - [The usefulness of a new class antihypertensive drug, angiotensin II receptor antagonist, for essential hypertension]. AB - Nonpeptide angiotensin II type 1-receptor antagonists, AT1 receptor antagonists, are newly developed and useful drugs for essential hypertension. In Japan, the efficacy and safty of losartan and candesartan cilexetil in patients with essential hypertension have been evaluated by the double-blind, parallel group comparison study using enelaprol maleate as control drug. Both trials revealed that these drugs showed a hypotensive effect comparable to that of enalapril with a high safety since the adverse drug reaction of cough was recognized in very few patients. Since the blood pressure normalizes only in the patient of about 50%, it is often required to add low-dose hydrochlorothiazide or calcium antagonist. Combination therapies further decreased blood pressure without any increases in side effects of the drugs. AT1 receptor antagonists in both mono-therapy and combination therapy with diuretics/Ca antagonists are very useful and safe in the hypertension treatment. PMID- 10361449 TI - [Efficacy of angiotensin II receptor antagonists as a novel drug for the treatment of chronic heart failure--in comparison with ACE inhibitors]. AB - Since 1) renin-angiotensin-aldosterone systems play an critical role in the development and progression of chronic heart failure, and 2) inhibitors of angiotensin converting enzyme (ACEIs) are proved to be effective for the treatment of chronic heart failure, angiotensin II receptor antagonists may be more effective than ACEIs. This is because angiotensin II receptor antagonists can inhibit the effects of angiotensin II via ACE-independent pathways, e.g., chymase. On the other hand, ACEIs can increase bradykinin, and thus, nitric oxide, which may cause potent cardioprotection. Therefore, angiotensin II receptor antagonists and ACEIs may mediate cardioprotection via different mechanisms, which may hint the combination therapy of both drugs in the pathophysiology of chronic heart failure. Angiotensin II receptor antagonists may open a new era for the treatment of chronic heart failure. PMID- 10361450 TI - [Vascular remodeling and angiotensin II]. AB - Numerous pharmacological approaches have failed to modify the high incidence of restenosis after balloon coronary angioplasty. This inability to alter the restenosis process was caused in part by our incomplete understanding of its pathology. Neointimal formation and geometric remodeling cause the restenosis after angioplasty. The development of restenosis involves vascular smooth muscle cells (VSMCs) migration and proliferation. Angiotensin II has been shown to stimulate the growth of VSMCs via an action on the angiotensin II AT1 receptor subtype. Angiotensin II AT1 receptor antagonists can block the angiotensin II induced these actions. Angiotensin II AT1 receptor antagonists may block the restenosis after balloon coronary angioplasty. PMID- 10361451 TI - [Potential role of angiotensin receptor antagonists in renal protection]. AB - Recent clinical studies have established an important role of angiotensin converting enzyme inhibitors (ACE-I) as a tool for renal protection. Although angiotensin receptor antagonists (AII-A) share the common property with ACE-I with regard to blockade of angiotensin activity via angiotensin type 1 receptors (AT1), AII-A is also reported to stimulate AT2 that plausibly activates nitric oxide production within renal medulla and augments synthesis of vasodilatory P450 metabolites in renal afferent arterioles. In contrast, AII-A is reported to have no effect on bradykinin activity. Results obtained in experimental animals indicate that AII-A effectively prevents the progression of renal injury. Several clinical studies are in progress, and the preliminary results suggest that AII-A has potent renal protective action in a variety of renal disorders. PMID- 10361452 TI - [Angiotensin and arteriosclerosis: an approach from transgenic and knockout mice]. AB - This brief review was designed to summarize the relationship between the renin angiotensin system and arteriosclerosis. Tsukuba hypertensive mice (THM) carrying both the human renin and angiotensinogen genes and C57BL/6J control mice 2 to 3 months of age were fed with either a western or normal diet for 14 weeks. Compared with controls, microscopic analyses revealed accelerated damage of cellular structure in the aortic root in THM fed with the western diet. Remarkably, the surface area of arteriosclerotic lesion in THM was shown to be 4 times larger than that in C57BL/6J on the same western diet. These findings suggested that hypertension induced by the activated renin-angiotensin system is involved in the development of arteriosclerotic lesions. PMID- 10361453 TI - [The effects of angiotensin II receptor antagonists on insulin resistance]. AB - Many of hypertensive individuals have glucose intolerance and dyslipidemia, and insulin resistance is common disorder on the basis of these diseases. It is important to take care of these metabolic disease for not only the control of hypertension, blood glucose and hyperlipidemia, but also the prevention of atherosclerosis. We reviewed the effects of angiotensin II receptor antagonists on insulin resistant syndrome. PMID- 10361454 TI - [Comparison with other antihypertensive drugs, especially with ACEI]. AB - In this review angiotensin II receptor antagonists (Angiotensin antagonists are discussed on the efficacy and safety in the treatment of essential hypertension. Angiotensin antagonists are more complete renin angiotensin system blockade, and are potent as ACE inhibitors, but they have rarely troublesome dry cough specific to ACE inhibitors. Angiotensin antagonists have demonstrated an excellent tolerability profile. Angiotensin antagonists will have potentially greater protection from end-organ damage, since they have provided end-organ protection in animal experiments. These agents will be considered for first-line therapies in very near future. PMID- 10361455 TI - [Drug-interactions and adverse effects of losartan potassium, an angiotensin II receptor antagonist]. AB - Losartan potassium is mainly metabolized by P450 chiefly in the liver. A P450 inducer, phenobarbital, has no significant effects on the pharmacokinetics of losartan. Cimetidine, known to inhibit P450 activity, has no remarkable effects on the metabolism of losartan. Side effects of losartan has been very few in the clinical trials of this drug on several thousands of patients so far. The rate was as low as that of placebo. Losartan may cause hyperkalemia when used with potassium-sparing diuretics, such as spironolactone or triamterene. Angioedema and acute hepatitis had been reported in 3 patients among thousands of millions of hypertensives under losartan treatment. The etiology was unclear, simple coincidence may not be ruled out. Losartan should not be administered to pregnant women and breast-feeding mothers, because it may disturb the fetal growth or may be harmful to the newborn. It should be cautiously used in patients with renal failure or liver disfunction. PMID- 10361456 TI - [Estrogen receptor and selective estrogen receptor modulators (SERMs)]. AB - The incidence of osteoporosis and of cardiovascular disease increases in women after menopause. Although theses diseases can be prevented by estrogen replacement therapy, this treatment is associated with an increased risk of endometrial cancer and perhaps also with an increased risk of breast cancer. Thus, a therapy that could prevent postmenopausal bone loss and lower serum cholesterol concentrations without stimulating reproductive tissues would be desirable. Selective estrogen receptor modulators (SERMs), such as raloxifene and tamoxifen, produce beneficial estrogen-like effects on bone and lipid metabolism, while antagonizing estrogen in reproductive tissue. Both agonist and antagonist activities are mediated via high affinity interaction with the estrogen receptor (ER). Both types of ER (alpha and beta) may be involved in the mechanism by which SERMs produce tissue-selective pharmacology. This review will discuss the roles of ER alpha and ER beta in novel signal transduction pathways. PMID- 10361457 TI - [Implantation of the artificial retina]. AB - In some degenerative retinal diseases, e.g., retinitis pigmentosa and age-related macular degeneration, the photoreceptors are destroyed to cause serious visual defects. Recent studies on blind human subjects revealed that a large number of ganglion cells remains intact and is capable of transmitting signals to the brain to evoke partial visual perception. This provided hope to compensate for the visual defects with retinal prostheses. The recent progress of microfabrication technique made it possible to implement the Vary Large Scale Integrated circuit, the artificial retina, which emulates a part of retinal function. The idea of implanting the artificial retina to the patients was proposed recently and experiments using animals have been put into practice. This article surveys the front line of the artificial retina implantation. PMID- 10361458 TI - [Health and pain]. PMID- 10361459 TI - [Some relations between pain and identity]. PMID- 10361460 TI - [Migration and pain. Need for epidemiologic research]. PMID- 10361461 TI - [Homo dolens. Grievances of a generalist concerning the persistent-- said "somatoform"--pain syndrome]. PMID- 10361462 TI - [The provoking dimension of pain. From somatoform pain disorder to a phenomenological approach to illness]. PMID- 10361463 TI - [Certificates for disability insurance and for refugees: similarities]. PMID- 10361464 TI - [Homage to Jean Wertheimer]. PMID- 10361465 TI - [Biology, psychology stakes for the psychogeriatrics of tomorrow]. PMID- 10361466 TI - [Confused states in the elderly]. PMID- 10361467 TI - [Therapeutic approach to depressive disorders in the elderly person]. PMID- 10361468 TI - [For a new management of the life ages: example of the age of retirement]. PMID- 10361469 TI - [Progress achieved in the prevention and therapy of Alzheimer's disease]. PMID- 10361470 TI - [The use of screening tools in psychiatry of the elderly person]. PMID- 10361471 TI - [Early decline in an epidemic: evolution of the prevalences of HIV, hepatitis B, hepatitis C and aminotransferases in intravenous drug abusers in Strasbourg between 1980 and 1990]. AB - OBJECTIVE: In order to study the prevalence of hepatitis B and C as well as HIV virus and disruptive factors of aminotransferase enzyme among intravenous drug users. METHOD: We conducted retrospective study of 169 patients who were hospitalised in a clinic where they were weaned from drug use between 1980 and 1990. RESULTS: The prevalence of HIV is 13.2%, and that of hepatitis B is 67.5%. When comparing the years 1980-1987 and 1987-1990, we noticed that the prevalence of HIV drops from 25.9% to 7.8% (p = 0.006), and that of hepatitis B from 75.9% to 60% (p = 0.027). Since November 1989, we were able to detect hepatitis C virus on a regular basis, and it showed a prevalence of 67.4%. The prevalence of aminotransferase enzyme disorders is 52.7%. CONCLUSION: We compare these results with those of other material on the same subject and we discuss various ideas to explain the unexpected decrease of HIV prevalence. PMID- 10361472 TI - [Hormones and voice or does Orpheus have an age?]. PMID- 10361473 TI - [Emergencies by telephone]. PMID- 10361474 TI - [Deceptive publicity]. PMID- 10361475 TI - [Homeopathic "point of view"]. PMID- 10361476 TI - The bird flu--what lies ahead? PMID- 10361477 TI - "Not to be ministered unto, but to minister". PMID- 10361478 TI - Case mix--for better or for worse? PMID- 10361479 TI - Consultation length and case mix in a general practice clinic. AB - OBJECTIVE: This study was conducted to determine the mean consultation length in a general practice clinic for all cases as well as for acute and chronic conditions. METHODS: The main diagnosis or reason for encounter in a general practice clinic from 25 April to 15 May 1994 was coded for all consultations using a customized clinic management software. RESULTS: The case mix of the clinic was comparable to the general practice pattern described in the 1993 Morbidity Survey of Outpatients. The overall mean consultation length was 9.3 minutes, the median was 6.0 minutes and the mode was 3.0 minutes. The mean consultation length for representative acute, chronic, and chronic relapsing and remitting conditions were 7.1 minutes (acute upper respiratory tract infection), 7.6 minutes (hypertension), and 9.9 minutes (bronchial asthma), respectively. CONCLUSION: Consultation length for a practice is dependent on the case mix of the practice, which is in turn dependent on the number of tasks required. PMID- 10361480 TI - An audit of endoscopic sinus surgery. AB - AIM OF STUDY: A prospective study was carried out to evaluate the results of endoscopic sinus surgery based on symptom score and endoscopic findings and to evaluate the prognostic factors using an audit form designed by IS Mackay of London. METHODS: Consecutive cases undergoing endoscopic sinus surgery were entered into the study. Pre-operative symptom, computer tomographic and endoscopic scores were recorded. Operations were also evaluated objectively; post operative symptom and endoscopic scores were recorded at 3, 6 and 12 months. Cases lost to follow-up were interviewed over the telephone and offered free endoscopic assessments. Those who required revision surgery were considered failures and re-entered into the study. RESULTS: The study comprised 113 cases (108 patients, 60 males and 48 females) with ages ranging from 14 to 80 years (mean 40.4). There were 52 with chronic rhinosinusitis, 46 with nasal polyps, 7 with tumours/inverted papillomata, 5 with acute complicated sinusitis and 3 with fungal sinusitis. Of these, 81.9% of patients with chronic rhinosinusitis or polyps showed an overall improvement of symptoms. Nasal obstruction showed the greatest improvement, followed by rhinorrhoea, loss of smell and headaches and facial pain. Endoscopic improvement was greatest in polyps (83.3%) and oedema (87.0%) but less for discharge (59.4%). CONCLUSIONS: Our results compared well with international series. Computer tomography scoring and the number of revision operations emerged as potential prognostic indicators requiring further evaluation. PMID- 10361481 TI - Congenital hypothyroid screening using cord blood TSH. AB - BACKGROUND: Clinical diagnosis of congenital hypothyroidism (CH) is difficult at birth without neonatal screening. In line with the priorities of the national health services in Malaysia towards preventive medicine, early diagnosis and treatment of CH is emphasised. We conducted a pilot study at Kuala Lumpur's Maternity Hospital between April 1995 and November 1995 to estimate the incidence of CH and also evaluated the problems associated with large-scale neonatal screening using a commercial TSH kit on cord bloodspots. PATIENTS: A total of 11,000 newborns were screened using cord blood spots taken at birth. RESULTS: Two hundred and fifty newborns (2.27%) hand cord TSH > 20 mlU/L and had to be recalled for re-evaluation. Of these, 4 had cord TSH of > 100 mlU/L; three were confirmed to have congenital hypothyroidism and one had transient hyperthyrotropinaemia. Our study estimated the incidence of CH to be one in 3,666 live births in Kuala Lumpur, Malaysia. Clinical features of hypothyroidism are subtle during the early weeks of life. However, prolonged neonatal jaundice (3/3), widely opened posterior fontanelle (3/3) and dry skin (3/3) were the common features in all our cases by 2-6 weeks of life. CONCLUSION: This study suffered a high dropout rate. Twenty-six percent of the patients were not traceable after discharge and 48% did not respond to our recall. We stress the importance of public education and awareness in contributing to the cost effectiveness of the screening program. PMID- 10361482 TI - Non-contrast high resolution fast spin echo magnetic resonance imaging of acoustic schwannoma. AB - AIM OF STUDY: The current gold-standard of examination for the exclusion of acoustic schwannomas is contrast-enhanced magnetic resonance (MR) imaging. Many patients however, still cannot afford to pay for the cost of this examination. As a result, many clinicians still resort to contrast-enhanced computed tomography (CT) scan; an examination which could miss small intracanalicular acoustic schwannomas. The aim of this study was to report on our experience on the usage of the more affordable high-resolution fast spin-echo (FSE) MR imaging for the diagnosis of acoustic schwannoma. METHODOLOGY: A study involving 123 patients with symptoms of sensorineural hearing loss, vertigo and tinnitus was carried out between August 1996 and March 1997. All cases were scanned with a 1.5 T MR unit using a quadrate head coil. The section thickness was 1.5 mm, with TR/TE of 4000/96. Any mass arising from the vestibulocochlear nerve was considered to be an acoustic schwannoma, and most of these positive cases underwent further contrast-enhanced MR imaging to confirm the diagnosis. RESULTS: A total of 7 acoustic schwannomas were detected on high resolution FSE MR imaging and for 5 of these cases, the diagnosis was confirmed with contrast-enhanced MR imaging. Three of the 7 positive patients had contrast-enhanced CT scan done just prior to the MR study and the tumours were not detected on CT scan. The smallest acoustic schwannoma detected on high-resolution FSE MR imaging had a dimension of 0.5 x 0.4 x 0.5 cm. CONCLUSION: High resolution FSE MR imaging is more sensitive than contrast-enhanced CT scan in the diagnosis of acoustic schwannoma. Although the sensitivity is less than that of contrast-enhanced MR imaging, high resolution FSE MR is more affordable and therefore can play a role in the screening for acoustic schwannomas in selected groups of patients. PMID- 10361483 TI - A study of newly diagnosed epilepsy in Malaysia. AB - BACKGROUND/AIM OF STUDY: To determine the characteristics of newly diagnosed epilepsy in the multiracial population of Malaysia. METHODS: This is a prospective study of 165 consecutive newly diagnosed cases of epilepsy presenting to the neurology laboratory of the University Hospital, Kuala Lumpur. The inclusion criteria were: two or more seizures with interval of > 24 hours, age > 1 month, residents of Klang Valley. All the patients underwent an awake and sleep EEG. RESULTS: One hundred and sixty-five cases were collected over 1992-1994. Their ethnic origin was: Chinese (36%), Indian (35%), Malay (29%). The mean age of onset of epilepsy was 18.7 years. Localisation related epilepsies accounted for 57.6% of cases while the remaining 42.4% were generalised epilepsies. Of the generalised epilepsies, subclassification was as follows: idiopathic generalised epilepsy 28.5%, juvenile myoclonic epilepsy 5.5%, childhood absence epilepsy 3.6%, West syndrome 3%, Lennox Gastaut syndrome 1.2% and photosensitive epilepsy 0.6%. Twenty-two percent of the cases were symptomatic and 78% were cryptogenic/idiopathic. The patients had a mean of 3.9 other siblings. Only 0.76% of the close relatives (parents and siblings) had a history of epilepsy. CONCLUSION: The characteristics of epilepsy in Malaysia is largely similar to those reported elsewhere. Genetic factors may be playing a relatively minor role in causing epilepsy in this community. PMID- 10361484 TI - Management of adenocarcinoma in situ (ACIS) of the uteri cervix--a clinical dilemma. AB - OBJECTIVE: We retrospectively reviewed 24 cases of adenocarcinoma in-situ (ACIS) of the cervix, managed at KK Hospital, with the objective of determining our local approach to its treatment, the consequent clinical outcome and problems encountered. METHODS: Except for one case, all patients were treated between 1991 1996. Nineteen cone biopsies (17 laser and 2 cold knife) and eleven hysterectomies were performed. The mean follow-up duration was 20.5 months (range: 1-75 months). RESULTS: The mean age was 44.2 years (range: 32-68) with 80% of the cohort being more than 35 years old and the mean parity was 2.2. Six (25%) patients were symptomatic. Majority (21/23) had an abnormal initial Pap smear. Glandular lesions were found in 39% (9/23) of Pap smear, 28% (6/21) of colposcopy, 58% (8/14) of cervical biopsies and in 3 of 4 endocervical curettage. Of the 17 laser cone biopsy specimens, lesion involved the surgical margin in 6 patients (35%). ACIS was found in conjunction with CIN in 14 patients (58%). Five hysterectomies were performed for involved surgical margin and one for dubious surgical margin of the prior cone biopsy, of which 3 had residual ACIS. At the time of the study, there was no case of recurrent ACIS or overt adenocarcinoma developing following cone biopsy. CONCLUSION: Preconisation diagnosis of ACIS using Pap smear, colposcopy and cervical biopsy was found to be difficult in our series. Concurrent CIN occurred in a sizeable portion of patients. Laser cone biopsy was the preferred method employed. Total hysterectomy was frequently employed following cone biopsy for treating possible residual disease. We recommend greater vigilance for this condition especially in patients with CIN and the need for regular endocervical sampling in the follow-up of patients treated by cone biopsy. PMID- 10361485 TI - Clinical evaluation of risperidone in Asian patients with schizophrenia in Singapore. AB - OBJECTIVE: To evaluate the short-term efficacy and safety of risperidone in a group of Asian patients with schizophrenia in an 8-week open-label, prospective study. METHODS: Patients with DSM-IV schizophrenia were recruited from Woodbridge Hospital. After a washout period, they were started on a 56-day trial of risperidone. Outcome was assessed with the positive and negative syndrome scale (PANSS), the clinical global impression scale (CGI) and the extrapyramidal symptom rating scale (ESRS). RESULTS: The mean daily risperidone dose at end point was 5.6 mg (range, 3 to 8 md/day). Mean PANSS scores were reduced significantly from 78 +/- 15.1 at baseline to 56.6 +/- 10.9 at end point. Seventeen patients (85%) who were treatment responders, showed at least a 20% reduction in total PANSS scores at end point while nine patients (45%) had a greater than 50% reduction in the total PANSS scores. According to the CGI scale, 85% improved at end point. The severity of extrapyramidal symptoms (mean ESRS scores) were significantly lower at end point than at baseline. CONCLUSIONS: Risperidone was effective in the treatment of positive and negative symptoms of schizophrenia. PMID- 10361486 TI - Cutaneous tuberculosis mimicking cellulitis in an immunosuppressed patient. AB - A 28-year-old lady suffering from systemic lupus erythomatosus (SLE) with diffuse proliferative glomerulonephritis (DPGN) and who was on oral cyclophosphamide and prednisolone presented with left lower limb 'cellulitis'. The 'cellulitis' of the left lower limb failed to respond to usual antibiotics which prompted evaluation of the clinical diagnosis. The diagnosis is made based on the presence of granulomas, multinucleated giant cells and acid fast bacilli on the skin biopsy. PMID- 10361487 TI - Co-existing left atrial thrombus and myxoma in mitral stenosis--a diagnostic challenge. AB - We report an unusual case of an adult who underwent a mitral valve replacement with concomitant excision of the left atrial myxoma and thrombus. Echocardiography showed the presence of a large "thrombus" within the left atrial appendage, body and atrial septum. There was difficulty in trying to distinguish between the atrial thrombus and myxoma due to their morphological similarities. At time of surgery, frozen section confirmed the atrial septal component of the thrombus to be an atrial myxoma and the atrial septum was excised to obtain a clear margin. PMID- 10361488 TI - Stroke in Singapore--an overview. AB - Stroke is Singapore's third leading cause of death. The number of deaths and admissions to Singapore hospitals for stroke has been rising; when standardised for age, however, mortality rates for stroke for both genders have fallen. There has been a fall in the prevalence of hypertension, smoking and hyperlipidemia in the general population, with a rise in the prevalence of diabetes mellitus. The frequency of haemorrhagic stroke is higher than among Caucasian populations. While stroke patients tend to arrive to hospital early, the level of stroke awareness among stroke patients is poor. Inpatient and outpatient rehabilitation services are available. Many stroke patients are still disabled after their stroke. A national-level support group has been established. Over the years, the number of stroke victims and disabled stroke survivors will continue to rise. There is a need to persist with public education programs and risk factor screening, and to further develop hospital and community resources to meet this challenge. PMID- 10361489 TI - Clinics in diagnostic imaging (33). Missed testicular torsion. AB - A 13-year-old boy presented with a painful scrotal swelling. On examination, the left testis was enlarged and tender. Ultrasound scan showed diffuse hypoechogenicity, with absent intra-testicular but increased peri-testicular blood flow. The diagnosis of missed testicular torsion was confirmed at surgery. The role of imaging in differentiating among other causes of painful scrotal swelling, such as infection, trauma and tumour, are discussed. PMID- 10361490 TI - Abnormal electrocardiographic patterns in renal failure. Ventricular tachycardia. AB - A 9-year-old girl was diagnosed to have renal failure. However, her family has not been compliant with medical treatment and opted for traditional therapy instead. She was admitted in an ill state with fluid overload. What is the ECG diagnosis? PMID- 10361491 TI - What you need to know--hearing loss and inner ear diseases--can they be cured? PMID- 10361492 TI - Uricase-catalyzed oxidation of uric acid using an artificial electron acceptor and fabrication of amperometric uric acid sensors with use of a redox ladder polymer. AB - Electrochemical oxidation of uric acid catalyzed by uricase (uric acid oxidase, UOx; EC 1.7.3.3) was studied using several redox compounds including 5 methylphenazinium (MP) and 1-methoxy-5-methylphenazinium (MMP) as electron acceptors for UOx, which does not contain any redox cofactor. It was found that MP and MMP were useful to mediate electrons from UOx to an electrode in the enzymatic oxidation of uric acid. A novel redox polymer, poly(N-methyl-o phenylenediamine)(poly-MPD), containing the MP units was also found to possess the mediation ability for UOx, and poly-MPD was immobilized together with UOx onto an electrode substrate covered with a self-assembled monolayer of 2 aminoethanethiolate with use of glutaraldehyde as a binding agent. The resulting electrode (poly-MPD/UOx/Au) exhibited amperometric responses to uric acid with very fast response of approximately 30 s, allowing reagentless amperometric determination in a concentration range covering that in the blood of a healthy human being. Kinetic parameters of the apparent Michaelis constant and the maximum current response obtained at the poly-MPD/UOx/Au suggested that electrochemical oxidation of uric acid was controlled by diffusion of uric acid into the enzyme film and that the redox polymer worked well in mediating between active sites of UOx molecules and the electrode substrate. PMID- 10361493 TI - Sol-gel thin-film immobilized soybean peroxidase biosensor for the amperometric determination of hydrogen peroxide in acid medium. AB - An acid-stable soybean-peroxidase biosensor was developed by immobilizing the enzyme in a sol-gel thin film. Methylene blue was used as a mediator because of its high electron-transfer efficiency. The sol-gel thin film and enzyme membrane were characterized by FT-IR, and the effects of pH, operating potential, and temperature were explored for optimum analytical performance by using the amperometric method. The H2O2 sensor exhibited a fast response (5 s), high sensitivity (27.5 microA/mM), as well as good thermostability and long-term stability. In addition, the performance of the biosensor was investigated using flow-injection analysis (FIA). PMID- 10361494 TI - Electrochemical sensor for measurement of urea and creatinine in serum based on ac impedance measurement of enzyme-catalyzed polymer transformation. AB - Enzyme-catalyzed polymer transformation with electrochemical ac impedance detection has been employed for the measurement of urea and creatinine in serum samples. A polymer, based on poly(methylvinyl ether)/maleic anhydride modified by esterification with n-octanol, which is stable at pH 7.4 and which is transformed rapidly in response to alkaline pH changes, was linked to enzymatic reactions between urease and urea or creatinine deiminase and creatinine to produce a disposable sensor system. The polymer was screen-printed onto interdigitated screen-printed carbon electrodes and the electrodes overlaid with absorbent pads containing the relevant enzyme. Application of serum samples, "spiked" with either urea or creatinine, resulted in rapid polymer transformation, and resultant changes in the capacitance of the polymer-coated electrodes were analyte-concentration dependent. Additional information on the mechanisms of polymer transformation was obtained from dynamic quartz crystal microbalance measurements. PMID- 10361495 TI - Simultaneous enantiomeric determination of dansyl-D,L-phenylalanine by fluorescence spectroscopy in the presence of alpha-acid glycoprotein. AB - Few techniques are amenable to real-time analysis of enantiomers. In this paper, total complexation by alpha-acid glycoprotein (AGP) is shown to discriminate between enantiomers of dansyl-D,L-phenylalanine (DPs) by changing the local environment of the D and L enantiomers (DDP and DLP, respectively) from hydrophilic to hydrophobic. DDP and DLP show the same native fluorescence at lambda ex/lambda em = 200/544 nm in the absence of AGP, but show shifted emissions with a component at lambda ex/lambda em = 220/497 nm in the presence of AGP and in lipophilic solutions. The conditions for an analytical determination have been optimized, and the method has been used to measure the enantiomeric composition of DDP/DLP mixtures with concentration ratios varying over 2 orders of magnitude. The mechanism of chiral recognition for DDP and DLP by AGP is discussed and should be equally applicable to other dansyl-derivative amino acid enantiomers. The association constants for AGP with DDP and with DLP have been determined to be 1.33 x 10(2) L g-1 and 2.29 x 10(2) L g-1, respectively. PMID- 10361496 TI - Colloidal gold filtrates as metal substrates for surface-enhanced infrared absorption spectroscopy. AB - A new method for obtaining surface-enhanced infrared absorption (SEIRA) spectra of antibodies and antibody/antigen complexes has been developed. Antibodies attached to colloidal gold particles and then collected by filtration onto porous polyethylene membranes show enhanced spectral bands at 1080 and 990 cm-1 regardless of the antibody specificity. Attachment of a model antigen, glucose oxidase, to its specific antibody/colloid complex prior to collection produces enhanced bands at 1540, 1395, and 1250 cm-1. Similarly, when the antigen Salmonella is attached to its specific antibody/colloid complex prior to collection, a new enhanced band is observed at 1015 cm-1. Similarities and differences of the SEIRA spectra obtained on gold colloid are compared to previous work on gold films. PMID- 10361497 TI - Acetonitrile chemical ionization tandem mass spectrometry to locate double bonds in polyunsaturated fatty acid methyl esters. AB - A rapid method is presented for determining the location of double bonds in polyunsaturated fatty acid methyl esters (FAME) using an ion-trap mass spectrometer. The mass spectrum of the chemical ionization reagent acetonitrile in an ion trap includes a m/z 54 ion, identified previously as 1-methyleneimino-1 ethenylium ion. We show that it reacts with double bonds of polyunsaturated FAME to yield a series of covalent product ions all appearing at (M + 54)+. Collisional dissociation of these ions yields diagnostic fragments, permitting unambiguous localization of double bonds. For methylene-interrupted and conjugated FAME, one of these fragments results from loss of the hydrocarbon end of the chain, while the other involves loss of the methyl ester. Major diagnostic fragment ions for monoene and diene FAME occur as a result of cleavage adjacent to either allylic sites or double bonds in the original analyte and appear at one mass unit above the mass expected for homolytic cleavage. Fragmentation of polyene FAME yields major diagnostic ions resulting from cleavage between double bonds that appear one mass unit lower. The method is shown to produce highly characteristic spectra for FAME with 1 to 6 double bonds. Identification of double-bond position in highly unsaturated fatty acids is demonstrated in a mixture of unknown polyunsaturated FAME from an extract of cultured Y79 human retinoblastoma cells. PMID- 10361498 TI - Monitoring the growth of a bacteria culture by MALDI-MS of whole cells. AB - We have probed the time evolution of a growing bacteria culture by extracting samples periodically and performing matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) on whole cells. The mass spectra generated by this method contain tens of peaks in the 3-11-kDa mass range. Cultures of E. coli strain K-12 were grown in two types of containers and at two nutrient concentrations and sampled periodically from 6 to 84 h after inoculation. The relative intensities of several of the stronger peaks vary quite dramatically as a function of time. These temporal characteristics must be taken into account when MALDI-MS is applied to identify bacteria. The results also suggest that MALDI-MS can be used to follow the aging of a bacteria culture. PMID- 10361499 TI - Differentiation of diastereomeric N-acetylhexosamine monosaccharides using ion trap tandem mass spectrometry. AB - A quadrupole ion trap mass spectrometer equipped with electrospray ionization was used to distinguish three diastereomeric monosaccharides, N-acetylglucosamine, N acetylgalactosamine, and N-acetylmannosamine. The saccharides were derivatized to form the metal complex [CoIII(DAP)2HexNAc]Cl3 which, when collisionally activated, produced dramatically different product ion spectra. The product ion spectra generated for the three monosaccharide diastereomers were then used to confirm the stereochemistry of N-acetylhexosamines from a hydrolyzed oligosaccharide. Finally, the origin of each product ion was determined through isotopic labeling studies, and mechanisms were proposed which explain each resulting dissociation. PMID- 10361500 TI - A general method for producing bioaffinity MALDI probes. AB - A bioaffinity probe based on the idea of immobilizing avidin on the probe surface to extract biotinylated oligosaccharide is described. The probe is produced by taking advantage of the natural affinity of proteins for hydrophobic polymer films. The avidin is immobilized by simply drying the solution on a polymer film surface. This produces a bioaffinity probe that shows enhanced activity for biotin-labeled oligosaccharides. The probe is produced in a matter of minutes but is highly effective for concentrating biotinylated oligosaccharide on the surface. The best matrix for the analysis is DHB, and the best film for the probe is a polyester material commonly used for transparency film. The efficacy of the probe is illustrated with neutral and anionic oligosaccharides. Oligosaccharides derivatized with biotin are retained while those that are unlabeled are washed away. No trace of the unlabeled oligosaccharide is observed in the mass spectrum. PMID- 10361501 TI - A validated stable isotope dilution liquid chromatography tandem mass spectrometry assay for the trace analysis of cocaine and its major metabolites in plasma. AB - A validated method has been developed for the simultaneous quantitation of cocaine and its major metabolites (ecgonine methyl ester, benzoylecgonine, and norcocaine) in rat plasma. The method is based upon the use of stable isotope dilution liquid chromatography/atmospheric pressure chemical ionization/tandem mass spectrometry. Previously reported methods do not have the sensitivity and specificity that can be attained with this method. Plasma samples required no cleanup apart from protein precipitation, and no derivatization was required. Selected reaction monitoring was performed on the transitions of m/z 200 to m/z 182 (ecgonine methyl ester), m/z 290 to m/z 168 (benzoylecgonine), m/z 304 to m/z 182 (cocaine), and m/z 290 to m/z 168 (norcocaine). The standard curves were linear over the range from 2 ng/mL (benzoylecgonine, cocaine, and norcocaine) or 5 ng/mL (ecgonine methyl ester) to 1000 ng/mL in rat plasma. The lower limit of quantitation (LLQ) for benzoylecgonine, cocaine, and norcocaine was 2 ng/mL, and for ecgonine methyl ester, the LLQ was 5 ng/mL for plasma. This simple, rapid, reliable, and sensitive method of quantitation had excellent accuracy and precision for the four analytes. The method was sensitive enough to permit a detailed study of the pharmacokinetics of cocaine and its metabolites after administration of a bolus intravenous dose to rats. PMID- 10361502 TI - Analysis of common cold virus (human rhinovirus serotype 2) by capillary zone electrophoresis: the problem of peak identification. AB - Different preparations of human rhinovirus serotype 2 (HRV2), a common cold virus, were analyzed by capillary zone electrophoresis (CZE) in untreated fused silica capillaries using borate buffer (100 mmol/L, pH 8.3) and sodium dodecyl sulfate (10 mmol/L) as additive to prevent wall adsorption. The electropherograms showed one major peak at 205- and 254-nm detection wavelengths. The identity of the peak as originating from native virus was confirmed by several indirect methods. Heating to 56 degrees C is known to lead to release of the genomic RNA from the viral capsid; this treatment resulted in the disappearance of the major peak and the emergence of a new predominant peak that was identified as RNA by enzymatic digestion. As expected, RNase treatment of the unheated sample remained without effect as the viral genome is inaccessible in the native viral shell. A monoclonal, virus-aggregating antibody was used for immunodepletion of native virus; again, the major peak disappeared upon removal of viral aggregates by centrifugation prior to CZE analysis. In combination, these results allowed for the unambiguous identification of the main peak as native HRV2 and of the minor peaks as contaminants present in various amounts in the different viral preparations. It is demonstrated that CZE allows for an extremely easy and rapid assessment of conformational state and purity of virions in a given viral preparation. PMID- 10361503 TI - Electrophoretic separation of small DNA fragments in the presence of electroosmotic flow using poly(ethylene oxide) solutions. AB - A new and simple method was demonstrated for separating phi X-174/Hae III DNA restriction fragments and DNA markers V and VI, respectively, without filling capillaries with polymer solutions prior to analysis. Using this novel method, poly(ethylene oxide) (PEO) solutions containing ethidium bromide migrated into capillaries by electroosmotic flow (EOF) during the separation. Two DNA fragments (123 and 124 bp) in markers V and VI were well-resolved. RSD values for the separation of phi X-174/Hae III DNA restriction fragments were less than 0.52% for 3 runs using a single 75-micron capillary and less than 3.96% using three different 75-micron capillaries. A highly viscous polymer solution prepared from 3% PEO was also used for separation of DNA markers V and VI. Theoretical plates up to 11.91 million/m and separation times of less than 7 min were achieved in the separation of phi X-174/Hae III DNA restriction fragments using a 10-micron capillary and a 2% PEO solution. Advantages of this method include simplicity, short separation times, the ability to use highly viscous polymer solutions for separating small DNA fragments, and the possibility of introducing several different polymer solutions into capillaries to extend the DNA separation range. PMID- 10361504 TI - Symmetry breaking during the formation of beta-cyclodextrin-imidazole inclusion compounds: capillary electrophoresis study. AB - A mathematical model was developed for the estimation of binding constants by capillary electrophoresis. The effective electrophoretic mobility in a solute mixture is dependent on the cyclodextrin concentration in the background electrolyte (BGE) as well as the stoichiometry and the binding constant of the guest-cyclodextrin complex. As well, a determination of the degree of complexation, nc (the percent of complexed guest) could be carried out. The model was applied to a series of imidazole derivatives. Thermodynamic data for the solute complexation mechanism were calculated. Different van't Hoff plot shapes of the degree of complexation were observed with different BGE pH values, indicating a change in the solute complexation mechanism. Enthalpy-entropy compensation revealed that the solute complexation mechanism was independent of the imidazole derivative molecular structure, the same at pH = 4.5, 5.0, 5.5, 6.0, and 6.5 but changed at pH = 7.0 and 7.5. Topological defects formed during a symmetry-breaking transition could be responsible for the modification of the structure of the cyclodextrin cavity and explained the nc variations in relation to pH and temperature. PMID- 10361505 TI - Medical photographer's work documents treatment of severe polio. PMID- 10361506 TI - Heather Angel named 1998 Louis Schmidt Laureate. PMID- 10361507 TI - Use of peripheral blood blasts vs bone marrow blasts for diagnosis of acute leukemia. AB - Acute leukemia can be diagnosed when blasts constitute 30% or more of the nucleated cells in a patient's peripheral blood (PB) sample. To determine whether in such cases bone marrow (BM) aspirates are still necessary, we compared the results of diagnostic studies performed on PB samples with blast counts of 30% or more with those performed on the same patients' BM samples. We found no differences in morphologic features, cytochemistry, or immunophenotype between the blasts in PB and BM samples in any of 30 cases studied. However, in 10 (23%) of 44 cases in which cytogenetic analysis was performed, PB but not BM samples were insufficient for analysis. The converse never occurred. Five of the 10 cases had acute lymphoblastic leukemia and 5 had acute myeloid leukemia (41% of the patients with acute lymphoblastic leukemia and 17% of the patients with acute myeloid leukemia). In cases with adequate metaphases, there was strong correlation between the cytogenetic results for PB and BM samples. Some PB samples with blast counts of 30% or more are adequate for diagnosis of acute leukemia, especially when therapy can be delayed until it is known that an adequate number of analyzable metaphases are recovered from the PB samples. PMID- 10361508 TI - MUC1 expression in hematopoietic tissues. AB - The MUC1 gene encodes the core protein of episialin, which is recognized by several antibodies. Reverse transcription-polymerase chain reaction (RT-PCR) detection of MUC1 transcripts has been proposed for the detection of micrometastases from breast cancers. MUC1 expression in hematopoietic tissues has been reported but not confirmed. Our preliminary RT-PCR studies confirmed MUC1 expression by MDA-231 breast cancer cells. Western blots of MDA-231 proteins stained with anti-MUC1 core gave one 68-kd (core protein) band, with an additional high molecular weight (HMW) band in blots stained with anti-epithelial membrane antigen (EMA). MUC1 expression was detectable by RT-PCR in 4 samples each of peripheral blood, bone marrow, and lymph node. MUC1 expression was detectable by Western blot analysis using anti-MUC1 core and anti-EMA in 2 peripheral blood samples and all bone marrow samples. Western blots from all lymph node samples stained positively with anti-EMA for the HMW product, but the 68-kd product was less prominent. Separated peripheral blood lymphocytes and granulocytes showed similar levels of MUC1 expression. RT-PCR studies demonstrated MUC1 expression in various hematopoietic cell lines. Western blots showed the 68-kd and HMW products in a granulopoietic line, with only the 68-kd product in 3 lymphoblastoid lines. MUC1 is expressed ubiquitously in hematopoietic tissues and is unsuitable for use as a marker for epithelial micrometastases. PMID- 10361509 TI - A lysis, storage, and transportation buffer for long-term, room-temperature preservation of human clinical lymphoid tissue samples yielding high molecular weight genomic DNA suitable for molecular diagnosis. AB - Molecular diagnosis of malignant lymphoma depends on the ability to extract high molecular weight genomic DNA. However, collection, storage, and transportation of frozen tissue is time consuming and expensive. We used a simple, low-cost lysis, storage, and transportation buffer (LST) to maintain clinical tissue samples at room temperature for up to 4 weeks before molecular analysis. Immersion of lymphoid tissue in LST at room temperature for 2 to 4 weeks was compared with snap-freezing in liquid nitrogen followed by storage at -75 degrees C. Southern blot analysis using an immunoglobulin heavy chain JH probe yielded identical results in 5 clonal and 6 nonclonal samples. The DNA recovered from the LST of a 12th sample was too degraded to be analyzed; however, the tissue had large zones of geographic necrosis. We also demonstrated that DNA extracted from tissue stored in LST is suitable for amplification by the polymerase chain reaction. Results from 4 of the snap-frozen and LST samples analyzed for rearrangements at the immunoglobulin heavy chain VDJ locus were identical. LST can be used in a clinical laboratory for storing tissue samples at room temperature up to 4 weeks before molecular analysis. PMID- 10361510 TI - Comparison of touch imprints with aspirate smears for evaluating bone marrow specimens. AB - We compared the differential counts of normal and abnormal bone marrow from touch imprints with those from aspirate smears to determine whether the touch imprint was reliable for independent routine use in the examination of bone marrow and the classification of hematologic abnormalities. Normocellular bone marrow specimens were obtained from 87 patients without hematologic abnormality. Abnormal bone marrow specimens were obtained from 173 patients with treated or untreated neoplastic hematologic disease, including acute myeloid leukemia, myelodysplastic syndrome, chronic lymphocytic leukemia, non-Hodgkin lymphoma, hairy cell leukemia, myeloma, and acute lymphoblastic leukemia. We found no diagnostic difference in the differential counts from touch imprints and aspirate smears of normocellular bone marrow, and although we found some difference between the differential counts in certain cases of diseased bone marrow, the touch imprint proved to be a reliable diagnostic tool for determining the cellular composition of normal bone marrow and more reliable for the diagnosis of bone marrow involved by a neoplastic hematologic disease. Our findings suggest that evaluating touch imprints should be considered a standard practice in examining bone marrow. PMID- 10361511 TI - Posttherapy surveillance of B-cell precursor acute lymphoblastic leukemia. Value of polymerase chain reaction and limitations of flow cytometry. AB - Flow cytometric immunophenotypic analysis is critical in diagnosis and classification of acute leukemia and has been used after therapy to monitor for minimal residual disease. However, the presence of normal B-cell precursors, hematogones, particularly in the context of treated pediatric B-cell precursor acute lymphoblastic leukemia (BP-ALL), may confound such evaluation. In this study, the value of more specific genotypic markers (polymerase chain reaction evaluation of 2 antigen receptor genes) was assessed to resolve this issue. Flow cytometric analysis of enriched mononuclear cells revealed 1% to 20% precursor B cells (PBCs), based on expression of 1 or more pan-B cell antigens in addition to CD10, CD34, and terminal deoxynucleotidyl transferase in all 14 patients studied. Inasmuch as this mimicked the immunophenotype of the original leukemic clone, PBCs, in isolation, were considered suspicious for minimal residual disease. However, 11 of the 14 posttherapy specimens (79%) revealed no monoclonally rearranged antigen receptor genes, and 7 of these 11 patients had trackable genotypic markers at presentation. Accordingly, by PCR these 7 patients had complete molecular remission, supported by clinical follow up of 16 to 73 months. Among the remaining 4 patients with PCR-negative disease, 3 continue in remission, confirming the interpretation of false-positive flow cytometric analysis. In conclusion, flow cytometric monitoring of posttherapy bone marrow specimens from patients with BP-ALL may be misleading, if considered in isolation, in falsely suggesting the presence of minimal residual disease. Rather, PCR for antigen receptor gene rearrangements is a valuable and specific tool, helpful in differentiating hematogones from minimal residual disease in patients with treated BP-ALL whose bone marrow harbors increased PBCs. PMID- 10361512 TI - Acquired activated protein C resistance in postmenopausal women is dependent on factor VIII:c levels. AB - Activated protein C (APC) resistance is an established risk factor for venous thromboembolism. In 5% to 10% of patients with venous thromboembolism, the APC resistance phenotype is observed in the absence of factor V Leiden mutation. Moreover, some physiologic and pathologic conditions are associated with an "acquired" APC resistance, not caused by the Leiden mutation, such as inflammatory diseases, pregnancy, or oral contraceptive therapy. Several studies have demonstrated the effect of menopause on the hemostatic system, but no data are available about APC resistance. We found a high prevalence of APC resistance in postmenopausal women, not associated with factor V Leiden mutation. The mechanism that underlies this acquired APC resistance may be related to the higher levels of factor VIII, which showed a strong inverse correlation with APC resistance, whereas no correlation was found between the normalized APC ratio, factor V levels, and protein S values. Higher levels of factor VIII correlated with a marker of coagulation activation, such as prothrombin fragments 1 plus 2. Therefore, to identify women receiving hormone replacement therapy who have a greater risk for deep venous thrombosis, the APC resistance coagulation test should be used instead of the genetic study. PMID- 10361513 TI - The feasibility of using blood bank-stored (4 degrees C) cord blood, unmatched for HLA for marrow transplantation. AB - The main purpose for storing large numbers of umbilical cord blood (CB) units by cryopreservation is to obtain a close HLA match for use in bone marrow transplantation. The use of partially matched or unmatched CB has been suggested, and publications about the success of 3 antigen mismatches have given some credence to this suggestion. Graft vs host disease still is considered a major barrier for successful CB transplantation. The cost per frozen CB unit of approximately $15,000 considerably limits its availability in developing countries. Eleven human umbilical cord specimens were stored in gas-permeable bags at 4 degrees C for up to 3 weeks. Clonal growth, replating efficiency in methylcellulose cultures, differential count, and flow cytometric immunophenotyping results were examined at intervals up to 21 days. Mixed lymphocyte cultures were evaluated on 13 similarly stored specimens at intervals up to 14 days. When plated at 1, 10, and 21 days, the combined percentage of the more primitive colonies increased on days 10 and 21. Replating efficiency of blast cell colonies when stem cell factor was added was 81.2% and 67.8% on days 10 and 21, respectively. When mononuclear cells were immunophenotyped, the mean percentage of CD34+ and CD117+ cells, considered primitive stem cell markers, increased significantly from day 1 to day 21. The ability of stored CB cells to respond to phytohemagglutinin or alloantigens decreased progressively from day 1 to day 14. By day 14, the reactivity of CB responder cells, in mixed lymphocyte cultures, to fresh allogeneic CB stimulator cells declined significantly. These findings suggest that CB can be stored in existing blood bank facilities and retain its hematopoietic potential for transplantation. Furthermore, it may be feasible to combine individual CB samples to provide a sufficient number of viable stem cells for transplantation, substantially expanding the number of potential recipients. PMID- 10361514 TI - Histologic transformation of polymorphous low-grade adenocarcinoma of salivary gland. AB - Polymorphous low-grade adenocarcinoma of salivary gland origin (PLGA) was initially described in 1984 and has since become an established clinicopathologic entity. Owing to the indolent nature of PLGA and its relatively recent description, the full clinicopathologic spectrum of this entity has not been elucidated fully. Transformation to a histologically different-appearing lesion or progression to a higher histologic grade has not been reported. We describe 2 PLGAs arising in the palate and associated with multiple locoregional recurrences that were treated with excision and radiation therapy. This was followed by histologic transformation to a higher grade neoplasm after 17 and 26 years, respectively. The histologic appearance after transformation was characterized by a predominantly solid and cystic growth pattern, nuclear atypia with prominent nucleoli, and foci of necrosis. High-grade transformation of PLGA may occur after a protracted clinical course with multiple recurrences of typical PLGA. The possible role of radiation therapy as an initiator of this transformation merits further study. Tumors with these histologic features should not be included under the rubric of typical PLGA. Segregation of these neoplasms will allow further study of their biologic potential, particularly with regard to possible increased rates of local recurrence and metastasis. PMID- 10361515 TI - Virtual microscopy and the Internet as telepathology consultation tools. A study of gastrointestinal biopsy specimens. AB - Telepathology (TP) is the practice of pathology at a distance using videomicroscopy and telecommunication tools. We explore the use of "virtual microscopy" techniques and the Internet as tools for TP gastrointestinal biopsy consultations. Thirty-five gastrointestinal biopsy specimens were photographed in Los Angeles by using a high-resolution digital camera, a light microscope, and a Pentium 166 microcomputer. Several (2-8) digital photomicrographs were collected at 40x or 100x optical magnification, using 2,700 x 3,400 pixel resolution. The photomicrographs illustrated all the tissue fragments present in 1 of the biopsy levels. They were saved in medium compression JPEG image format. These images can be magnified digitally up to 600% without visible degradation and scrolled at different magnifications on a video monitor, simulating examination under a light microscope. The images files (281 to 3,324 KB) were attached to e-mail messages containing patient information and sent through the Internet to Michigan for interpretation using a Power Macintosh 7100 system. The e-mail process was successful in 100% of instances; 2 files were corrupted owing to user error and had to be resent. Additional photos were requested in 1 case. In 33 of 35 cases, there was diagnostic concordance between the original and the TP diagnoses. The 2 discrepancies were due to diagnostic disagreement. This technology offers pathologists relatively inexpensive and effective tools for gastrointestinal TP consultations. PMID- 10361516 TI - Renal oncocytoma and chromophobe renal cell carcinoma. A comparison of colloidal iron staining and electron microscopy. AB - Chromophobe renal cell carcinoma (RCC) is characterized by diffuse cytoplasmic staining with Hale colloidal iron (HCI) and the presence of numerous microvesicles. The eosinophilic variant morphologically may resemble renal oncocytoma. The latter commonly shows focal cytoplasmic HCI reactivity, but microvesicles have not been previously reported. We examined 19 chromophobe RCCs and 28 oncocytomas for their HCI staining patterns. Electron microscopy was performed on 13 chromophobe RCCs and 10 oncocytomas. In all cases of chromophobe RCC, more than 75% of cells showed a diffuse cytoplasmic HCI positivity, whereas a variable proportion of cells in 20 oncocytomas showed focal cytoplasmic staining, in a perimembranous, apical, or perinuclear pattern. Ultrastructurally, chromophobe RCCs contained abundant microvesicles with varying numbers of mitochondria, whereas all oncocytomas contained abundant mitochondria with focal collections of microvesicles. The microvesicles, in perimembranous, apical, or perinuclear clusters or singly scattered throughout the cytoplasm, were morphologically indistinguishable from those in chromophobe RCCs. In most cases, the microvesicle location and HCI staining pattern correlated. Chromophobe RCC and oncocytoma have distinctive morphologic features that usually allow their recognition. In difficult cases, HCI staining and electron microscopy may help, but the presence of HCI positivity or microvesicles in an eosinophilic renal tumor does not rule out oncocytoma. PMID- 10361517 TI - Chronic peripheral separation of placenta. The significance of diffuse chorioamnionic hemosiderosis. AB - Diffuse nonmeconium-related pigment was observed in the chorioamnion of 36 of 1,023 placentas over 4 years and evaluated by iron staining. Stains were negative in 13 cases and positive in chorionic plate and membranes (diffuse chorioamnionic hemosiderosis [DCH]) in 23 cases (3/1,000 deliveries; 25/1,000 placentas). Gestational age at delivery was lower in DCH and was inversely proportional to the magnitude of iron staining. Placentas with DCH were more likely to show circumvallation, old peripheral blood clots, increased chorionic-villous macrophages, and green discoloration. To evaluate demographic, obstetric, and perinatal factors associated with DCH, 2 gestational age-matched controls were selected for each DCH case. Multiparity, smoking, and chronic vaginal bleeding all were increased significantly with DCH, while intrauterine growth retardation and oligohydramnios were increased but did not achieve statistical significance. Gestational hypertension and advanced maternal age were significantly decreased with DCH, and cocaine abuse was uncommon (3 cases). Long-term neurologic sequelae of DCH were evaluated in a separate series of gestational age-matched very-low birth-weight infants with and without neurologic impairment at 2 years of age. No increased risk of neurologic impairment was found in patients with DCH. PMID- 10361518 TI - Serum alpha-fetoprotein levels in patients with chronic hepatitis C. Relationships with serum alanine aminotransferase values, histologic activity index, and hepatocyte MIB-1 scores. AB - Patients with chronic viral hepatitis and cirrhosis often have elevated serum alpha-fetoprotein (AFP) values, the causes of which are unclear. We studied 81 patients with chronic hepatitis C and the relationships of serum AFP and alanine aminotransferase (ALT) values, hepatic histologic features, and hepatocyte proliferation activity scores. Twenty-two patients had nil to mild fibrosis, 34 had moderate fibrosis, and 25 had marked fibrosis-cirrhosis. The mean serum AFP value was significantly greater in patients with more fibrosis. Serum ALT values were slightly greater in the marked fibrosis-cirrhosis patient group. The differences in the HAI and in hepatocyte MIB-1 scores were not significant. Among all patients, increasing serum AFP values significantly correlated with increasing ALT values. However, there were no significant correlations with serum ALT or HAI and serum AFP values. There was no association between serum AFP values and immunohistochemical staining for AFP within hepatocytes. These results suggest that elevated serum AFP values are the result of altered hepatocyte hepatocyte interaction and loss of normal architectural arrangements. The presence of marked fibrosis or cirrhosis, a state of significant altered hepatocyte architecture, may be the underlying cause of increased serum AFP, rather than necrosis or active regeneration. PMID- 10361519 TI - Pituitary adenomas and granular cell tumors. Incidence, cell type, and location of tumor in 100 pituitary glands at autopsy. AB - Incidentally detected pituitary adenomas were investigated in 100 pituitary glands at autopsy to determine the number, cell type, and location of tumors, and the presence of coexisting granular cell tumors in the neurohypophysis. Pituitary glands were sagittally sectioned at 1.5-mm intervals in toto and embedded in 1 cassette to orient location of each tumor. Twenty-four pituitary glands harbored adenomas, most smaller than 3 mm and the largest 6 x 5 x 4 mm. Two pituitary glands contained double adenomas of immunocytochemically different cell types. Of the 26 adenomas, 10 had lactotrophs, 2 had mixed lactotrophs-somatotrophs, 1 had mixed lactotrophs-luteinizing hormone cells, and 12 were nonfunctioning. One adenoma with adenocorticotropic hormone cells was also detected. Thus 25 of 26 (96%) adenomas were either lactotrophic or nonfunctioning; this percentage is much higher than that of surgically resected tumors. Twenty-two tumors were contiguous with or adjacent to the capsule from which the adenomas originated. Nine granular cell tumors were noted in the neurohypophysis; 3 coexisted with pituitary adenomas. Fourteen additional cases revealed small granular cell nests. Thus the incidental finding of nonfunctioning pituitary adenomas is relatively common in adults (24% of cases in this study), and the coexistence of pituitary adenomas and granular cell tumors may suggest a possible histogenic connection between anterior and posterior pituitary tumorigenesis. PMID- 10361520 TI - Thymoma, atypical thymoma, and thymic carcinoma. A novel conceptual approach to the classification of thymic epithelial neoplasms. AB - Primary thymic epithelial neoplasms have been the subject of much controversy in recent years owing to the difficulties posed by these tumors for precise histopathologic typing and prognostication. A number of classification schemes using different terminology have been proposed, none of which has satisfactorily managed to address all the problems and concerns related to these tumors. We present a proposal for a novel approach to the histologic classification of primary thymic epithelial neoplasms that is based on morphologic features of differentiation. The principle behind this classification scheme is that the determination of the cytologic degree of atypia and the identification of the organotypical features of thymic differentiation may permit accurate classification of these neoplasms into 3 simple and reproducible diagnostic categories: thymoma, atypical thymoma, and thymic carcinoma. We further reiterate the traditional concept that tumor staging, not histopathologic typing, has a more crucial role for accurate and reliable prognostication for the better differentiated forms of these tumors. PMID- 10361521 TI - Risk assessment in breast conserving therapy. PMID- 10361522 TI - Cost effectiveness of AFB staining in tissue specimens. PMID- 10361524 TI - Changes from within: improving lifestyle habits using personality type. PMID- 10361523 TI - Dieting readiness test fails to predict enrollment in a weight loss program. PMID- 10361525 TI - The personality of dietetics. PMID- 10361526 TI - HCFA releases proposed rule for diabetes education. PMID- 10361527 TI - Federal role in nutrition education, research, and food assistance for women and their families. PMID- 10361528 TI - Dietary applications of the stages of change model. PMID- 10361529 TI - How can stages of change be best used in dietary interventions? PMID- 10361530 TI - Dietary advice to reduce fat intake is more successful when it does not restrict habitual eating patterns. AB - OBJECTIVE: To determine the relative and combined effects of dietary advice to lower fat intake and alter eating frequency on nutrient intake and plasma lipid levels in men with recently diagnosed hyperlipidemia. DESIGN: In this 4-week dietary intervention trial, a 7-day food diary was used in the week preceding the intervention and on the third week of the intervention to determine eating frequency. A 7-day dietary history and lipid profile (after subjects had fasted) were taken at baseline and at the end of the 4 weeks of dietary intervention. SUBJECTS: Eighty nonobese, free-living men with newly diagnosed primary hypercholesterolemia. INTERVENTION: Group 1: fat intake was reduced and eating frequency decreased. Group 2: Fat intake was reduced and eating frequency maintained. Group 3: Fat intake was maintained and eating frequency decreased. Group 4: Fat intake was maintained and eating frequency increased. MAIN OUTCOME MEASURES: Changes in lipid profiles of the subjects and evaluation of subjects' perception of the dietary advice they received. STATISTICAL ANALYSES PERFORMED: Baseline and intervention values were compared using a paired t test; 2-way analysis of variance was used across treatment groups. RESULTS: Dietary advice to lower fat intake significantly lowered total and low-density lipoprotein cholesterol levels; this advice also decreased intake of total fat and saturated fatty acids significantly, but less so in the group asked to restrict eating occasions to 3 per day. Advice to alter eating frequency was found to be difficult to follow. APPLICATIONS: Lipid-lowering dietary advice should be incorporated into the existing eating frequency pattern of each person. PMID- 10361531 TI - Sensory responses to fat are not affected by varying dietary energy intake from fat and saturated fat over ranges common in the American diet. AB - OBJECTIVE: To examine the effects of manipulating dietary fat in foods on sensitivity and hedonic response to fat in selected foods. DESIGN: Twenty subjects were randomly assigned to a sequence of three 8-week experimental diets (average American diet, step 1 diet, low-saturated-fat diet) that varied in energy from fat (37%, 30%, and 26%, respectively) and saturated fat (17%, 10%, and 6%, respectively). Subjects participated in sensory tests designed to assess their sensitivity to and liking for fat in several foods, before the study (baseline), after consumption of each diet, and after the study (washout). SUBJECTS/SETTING: Subjects were participants in the Dietary Effects on Lipoprotein and Thrombogenic Activity (DELTA) study. RESULTS: No significant differences were found among diets for difference thresholds (i.e., just noticeable differences) for fat in milk and pudding, ad libitum mixing of low- and high-fat samples of milk and soup, and hedonic scaling of fat concentrations in milk and muffins and of cheese, mayonnaise, hot dog, and pastry samples. APPLICATIONS/CONCLUSIONS: Within the dietary fat ranges and for the fat stimuli tested in this study, dietary fat as percentage of energy from fat and saturated fat was not a significant determinant of sensitivity to and/or liking for fat. Sensory factors should not be a barrier to the implementation of low-fat diets such as the step 1 and low-saturated-fat diets. PMID- 10361532 TI - The Diet Quality Index revised: a measurement instrument for populations. AB - OBJECTIVE: To evaluate a revision of the Diet Quality Index called the Diet Quality Index Revised (DQI-R). DESIGN: The original Diet Quality Index was revised to reflect current dietary guidance, to incorporate improved methods of estimating food servings, and to develop and incorporate measures of dietary variety and moderation. The scoring of the original scale was reversed in direction and expanded to a 100-point scale to improve interpretability. METHODS/SUBJECTS: Data from the 1994 Continuing Survey of Food Intakes by Individuals were used. A sample of 3,202 adults aged 18 and older contributed 2 days of dietary intake data based on 24-hour recalls for the development and revision of various components of the DQI-R. STATISTICAL ANALYSES: Pearson correlation analyses, ordinary least squares regression analyses, and a nonparametric test to determine trends across ordered groups were used. RESULTS: The mean DQI-R score for the 1994 sample was 63.4 of a possible 100-point score. Sample respondents were more likely to have met dietary guidance in the areas of dietary cholesterol (66.9% met goal) and iron intakes (59.6% met goal) relative to the Recommended Dietary Allowances but less likely to have met goals related to fruit servings (19.6% met goal), grain servings (23.1% met goal), and calcium intakes (16.6% met goal) relative to the Recommended Dietary Allowance. There is a statistically significant quantitative and qualitative improvement in all components of the DQI-R as one moves from the lowest grouping of scores to the highest. For example, persons with DQI-R scores less than 40 consumed 43.9% of energy from fat, 72% of the Adequate Intake for calcium, and 6.7% of the recommended servings of fruit per day. In contrast, those with DQI-R scores greater than 80 consumed 24.2% of energy from fat, 101% of the Adequate Intake for calcium, and 137% of the recommended servings of fruit per day. APPLICATIONS: The DQI-R reflects the dietary guidance principles of macronutrient distribution, moderation, variety, and proportionality. Although the index was designed to monitor dietary changes in populations rather than individuals, each index component reflects an aspect of national dietary guidance. Calculation of DQI-R scores for an individual should provide an estimate of diet quality relative to national guidelines, and differences in scores over time should suggest improvement or decline in overall diet quality. PMID- 10361533 TI - Innovative newsletter interventions improve fruit and vegetable consumption in healthy adults. AB - OBJECTIVE: To evaluate the effectiveness of computer-tailored newsletter interventions in improving the number and variety of fruits and vegetables eaten by adults. DESIGN: The 4-group randomized trial with pre- and postintervention measures consisted of a control group and 3 intervention groups receiving nontailored newsletters, computer-tailored newsletters, or tailored newsletters with tailored goal-setting information. Intervention groups received 1 newsletter each month for 4 months. SUBJECTS: Baseline surveys were completed by 710 health maintenance organization clients. Postintervention surveys administered 6 months after baseline were completed by 573 participants (80.8%). INTERVENTION: All newsletters contained strategies for improving fruit and vegetable consumption. Tailored newsletters used computer algorithms to match a person's baseline survey information with the most relevant newsletter messages for promoting dietary change. MAIN OUTCOME MEASURES: Daily intake and weekly variety of fruits and vegetables were measured using a food frequency questionnaire. STATISTICAL ANALYSES PERFORMED: Analysis of covariance and Tukey's honestly significant difference test were used to assess differences in the number and variety of fruits and vegetables consumed among intervention groups. RESULTS: For persons completing postintervention surveys (n = 573), all 3 newsletter groups had significantly higher daily intake and variety scores compared with the control group. Although there was a trend of improved intake and variety with each added newsletter element, there were no significant differences at follow-up among the newsletter groups. CONCLUSIONS: Newsletters can be effective in improving the fruit and vegetable consumption of adults. In this study, a computer-tailoring system did not significantly enhance the effect of the nutrition newsletters on fruit and vegetable intake. PMID- 10361534 TI - Dietary intake and body mass index of adults in 2 Ojibwe communities. AB - OBJECTIVE: To describe and compare dietary intake and prevalence of overweight in a sample of adults in 2 Ojibwe communities in Mille Lacs, Minn, and Lac Courte Oreilles, Wis. DESIGN: Cross-sectional survey based on interviews that included a 24-hour recall, food frequency questionnaire, and a sociocultural questionnaire. SUBJECTS/SETTING: One hundred four adult Band (tribe) members were selected randomly from current housing lists; pregnant and lactating women were excluded. STATISTICAL ANALYSES PERFORMED: Nonparametric Wilcoxon rank sum tests were used to determine differences in absolute nutrient intakes where normality could not be assumed. Two-tailed t tests were conducted to test for differences between nutrient densities. A significance level of alpha = .05 was used; procedurewise adjustments were made using the Bonferroni method when adjusting for multiple comparisons. RESULTS: The importance of the traditional food system was evident: at least 50% of respondents engaged in hunting and fishing practices. Prevalence of overweight was 47%. Mean nutrient intakes were below the Recommended Dietary Allowance for women for vitamin A, folate, calcium, iron, and zinc and for men for vitamin A and calcium, despite energy intakes that met the Recommended Dietary Allowance. Nutrient densities were lower than those in the third National Health and Nutrition Examination Survey for women for carbohydrate, vitamin C, folate, calcium, and dietary fiber and for men for folate and dietary fiber. Fat contributed 37% (for men) and 40% (for women) of energy intake. APPLICATIONS/CONCLUSIONS: Areas of focus for culturally relevant education programs (eg, promotion of traditional foods that provide nutrients of low intake status and low-fat traditional food preparation methods) and research needs are suggested to reduce risks for nutrition-related chronic disease among Native Americans. PMID- 10361535 TI - Using 3 data sources and methods to shape a nutrition campaign. AB - The first objective of this research was to define a target population of African American women more clearly. The second was to provide specific information about the needs and preferences of that population in order to design an effective, culturally relevant, community-based communications campaign to promote more healthful lifestyles. Data collection and analysis included the following: interviews with 10 community nutritionists and the director of the State Office of Nutrition, 6 focus groups with a total of 47 members of the target population, and direct observation and documentation of key community resources. This approach, called "triangulation," permits more in-depth understanding of issues, provides different perspectives on the problem, and helps ensure accuracy of conclusions. Interviews with nutritionists identified young African-American women as the appropriate target population for the campaign. These interviews and the focus-group discussions confirmed the acceptability of higher weight and better body-esteem among African-American women than among white women. Both the nutritionists and the focus-group members identified the need and desire for information and skills related to food preparation and provided specific direction for program content. Community observation confirmed the need for food markets with merchandise of consistently high quality, especially in the fresh and frozen produce sections. Observation also helped identify community services and programs. The 3 sets of data, which augmented a comprehensive literature review, provided a firm foundation for the campaign's design and development. Dietitians and nutritionists working in community settings can use triangulation to gain a better understanding of their populations in order to develop more effective interventions. PMID- 10361536 TI - Practitioners' guide to meeting the vitamin B-12 recommended dietary allowance for people aged 51 years and older. AB - In response to research findings that 10% to 30% of people aged 51 years and older may have protein-bound vitamin B-12 malabsorption, the National Academy of Sciences' Institute of Medicine recommends that these people consume a majority of the new Recommended Dietary Allowance (RDA) of 2.4 micrograms/day in its synthetic form rather than in its food form. Protein-bound vitamin B-12 malabsorption in older adults has been attributed to reduced pepsin activity and gastric acid secretion, which interfere with cleavage of vitamin B-12 from dietary protein before absorption. Unlike patients with pernicious anemia, most people with protein-bound vitamin B-12 malabsorption produce intrinsic factor and have the ability to absorb synthetic vitamin B-12 normally. Early diagnosis is necessary to prevent the untoward effects of vitamin B-12 deficiency. A thorough assessment of vitamin B-12 status entails measurement of multiple biochemical assessment indexes, including serum vitamin B-12, methylmalonic acid, and homocysteine concentrations. Dietitians and other health care professionals should be aware of the prevalence of vitamin B-12 deficiency in older adults and be familiar with sources of synthetic vitamin B-12 to facilitate implementation of the new RDA. PMID- 10361537 TI - Stages of change for reducing fat and increasing fiber among dietitians and adults with a diet-related chronic disease. PMID- 10361538 TI - Fat content of restaurant meals: comparison between menu and experimental values. PMID- 10361539 TI - Serving meals of reduced portion size did not improve appetite among elderly in a personal-care section of a long-term-care community. PMID- 10361540 TI - Ethnic differences in body image attitudes and perceptions among women infected with human immunodeficiency virus. PMID- 10361541 TI - Position of the American Dietetic Association and Dietitians of Canada: women's health and nutrition. AB - Within the past 10 years women's health has evolved to a much broader paradigm, beyond reproductive issues. From a physiological perspective, women's health now refers to the prevention, diagnosis, and management of conditions or diseases that may be unique to women, be more prevalent in women, or manifest differently in women than men. Women's health encompasses emotional, social, cultural, spiritual, and physical well-being. It is determined by the social, political, and economic context of women's lives. Nutrition is involved in the etiology or treatment of half of the 10 leading causes of death in women. The incidence of osteoporosis and extremes in body weight are approaching epidemic proportions in women. This position reviews the following health problems: cardiovascular disease, cancer, osteoporosis, weight, and diabetes mellitus. Dietetics professionals are in the perfect position to understand the issues surrounding women's health in order to deliver a message to women that will allow them to make wise decisions regarding their health. Nutrition is a critical component of risk reduction and treatment, and must be included in clinical and preventive services for women. Dietetics professionals must work to increase their knowledge about women's health issues, to promote health and education programs, to influence policy makers, to deliver the highest-quality medical nutrition therapy, and to be proactive in documenting the effectiveness of outcomes-based research. PMID- 10361542 TI - Recent advances in imaging of cerebrovascular disease. AB - The recent advances made in CT and MR imaging have led to increased accuracy in making a number of diagnoses in the emergency room setting. Increasingly, radiologists are asked to perform these studies and accurately interpret the findings, which often have a dramatic impact on triaging and treatment of the patient. Future trials need to address further the relative merits of each of the techniques outlined previously in specific settings. In addition, given the increasing number of means of obtaining diagnostic information, cost effectiveness studies are needed to better formulate an appropriate algorithm for each diagnosis. PMID- 10361543 TI - Helical CT and three-dimensional CT of facial and orbital injury. AB - Knowledge of the regions of the face and their buttresses and knowledge of the types of facial injuries frequently encountered simplifies the diagnostic task. The indications for CT include detection of suspected fractures and preoperative planning. The cost of facial CT to the hospital has declined and is little different than the cost of plain films. CT may become the screening modality of choice depending on the cost structure at any given hospital. PMID- 10361544 TI - Helical CT of cervical spine and soft tissue injuries of the neck. AB - Helical CT of the neck has revolutionized the diagnostic evaluation of trauma and emergency room patients. This comprehensive examination, with high resolution and fast acquisition times, allows the radiologist to make expeditious diagnoses concerning cervical spine fractures, vascular injuries, and aero-digestive tract lesions. This allows for the more rapid triaging and treatment of various injuries resulting in improved patient priate radiographic examination for each clinical scenario. PMID- 10361545 TI - Imaging diagnosis of nonaortic thoracic injury. AB - Chest radiographs remain the initial imaging modality to rapidly screen patients with blunt chest trauma. Spiral CT is more sensitive and specific in diagnosing most thoracic pathology seen in blunt trauma patients. This article reviews the major clinical and radiologic findings that occur with blunt injuries to the chest, excluding mediastinal vascular injuries. PMID- 10361546 TI - Helical CT grading of traumatic aortic injuries. Impact on clinical guidelines for medical and surgical management. AB - Helical CT is a reliable method for screening patients with blunt chest trauma for vascular and visceral injuries. Thoracic aortic injuries detected by CT examination affect the immediate clinical management and patient triage. This article describes the clinical indications and imaging protocols for helical CT of the chest used to detect aortic injuries, provides a grading system of the range of aortic injuries demonstrated by CT, and discusses the clinical management decisions that should be considered based on the CT grade of the aortic injury. PMID- 10361547 TI - Helical (spiral) CT in the evaluation of emergent thoracic aortic syndromes. Traumatic aortic rupture, aortic aneurysm, aortic dissection, intramural hematoma, and penetrating atherosclerotic ulcer. AB - For the near future, CT will play the critical and dominant role in the evaluation of patients presenting with emergent aortic syndromes. Its convenience, accuracy, and utility in the rapid evaluation of not just the aorta, but the entire thorax, make it ideally suited for use in emergency settings. Further benefits are likely to be realized in speed and resolution with multislice CT, although it is as yet not widely available. PMID- 10361548 TI - Helical CT of abdominal trauma. AB - CT has revolutionized the diagnostic work-up of trauma patients with suspected abdominal injuries. A wide range of intraperitoneal and retroperitoneal organ injuries can be quickly and accurately diagnosed with CT. Today, helical CT technology permits even faster examinations, with improved intravenous contrast opacification of parenchymal organs and vascular structures and reduced CT artifacts caused by patient motion, respiration, and arterial pulsation. Severely injured and potentially unstable patients, who might not have been able to tolerate the long CT examinations of the past, may be quickly evaluated today with helical CT. Accurate diagnosis requires high quality CT examinations that are performed with optimum CT protocols. This article reviews the currently recommended helical CT protocols for evaluating patients with suspected abdominal injuries, and the CT findings when injuries are present. PMID- 10361549 TI - The biology of hormone refractory prostate cancer. Why does it develop? AB - Androgen ablation therapy has been an important modality for the treatment of disseminating prostatic cancer for nearly 60 years. Unfortunately, when given alone, such therapy is rarely curative. The failure of this therapy to cure prostate tumors, even though it can induce an initially positive response, is not the result of a change in the systemic effectiveness of such treatment. Instead, the development of resistance to therapy is related to changes in the tumor. Experiments by a large number of investigators have identified several of the important tumor cell and host factors involved in these changes. Through the identification of these factors, the concept has evolved that there may be multiple pathways for the development of resistance to hormonal therapy based on a stem cell model for the normal prostate. Although such pathways can be described in phenomenological terms, the detailed molecular biology of such a process is still unknown. The essential feature of the development of androgen resistance is the emergence of androgen-independent or sensitive cancer cells. The critical question for that must be answered by future studies is exactly how such androgen-independent cells develop. An explanation may make it possible to design therapies to prevent the development of these independent tumor cells. Under such conditions, androgen ablation therapy used as a single modality could become potentially curative. Even if therapeutic means can be developed to prevent the emergence of androgen-independent or sensitive tumor cells, to be effective, this type of blocking therapy would have to be performed before such development had already occurred. Therefore, before such therapy is begun, some type of clinical test would be required to determine that the tumor did not already have some androgen-independent or sensitive tumor cells present (i.e., the tumor was not already heterogeneous androgen-sensitive). Because, currently, neither a method for determining the homogeneous versus heterogeneous nature of the androgen requirements of a particular tumor nor a method for the prevention of the development of androgen-independent or sensitive tumor cells from dependent prostate cancer cells is available, these should be critical areas for extensive future study. Any advancement in either of these areas would have profound consequences on the more effective issue of androgen ablation therapy. Until these advancements are made, androgen ablation therapy can be used in combination with other modalities of treatment (e.g., radiation and chemotherapy), which are specifically targeted at the androgen-independent or sensitive cells either initially present or developing during androgen ablation therapy. Standard antiproliferative chemotherapeutic agents may be ineffective against such androgen-independent or sensitive prostatic cancers because these cancers have a low proliferative rate. Berges and co-workers demonstrated that the median daily proliferative rate of prostate cancer cells within lymph nodes or bone metastases was less than 3.0% per day. Newer agents are needed to target the greater than 95% of prostate cancer cells within a given metastatic site that are not immediately proliferating. One such approach that has been recently proposed is the use of potent and selective inhibitors of the endoplasmic reticulum Ca2+ ATP-dependent pump. In such combination approaches, it will be critical to evaluate the importance of both the timing (early versus late) and the order (sequential versus simultaneous) of androgen therapy in relation to the other modalities used. PMID- 10361550 TI - Rational use of chemotherapy. It is not just rat poison. AB - The role of chemotherapy in the management of advanced prostate cancer is expanding. Multiple regimens that use a variety of drugs have been developed. A chemotherapy regimen may appear to be a collection of randomly chosen agents. This article presents the theoretic foundation for the development of combination chemotherapy. PMID- 10361551 TI - Serologic tumor markers, clinical biology, and therapy of prostatic carcinoma. AB - PSA has been a valuable tool in enhancing our understanding of the prevalence and virulence of prostate cancer. PSA also has contributed to the understanding of important phenomena related to the androgen regulation of the cancer; however, it has not been useful in detecting some forms of androgen-independent (neuroendocrine) progression and is of limited prognostic value in androgen independent prostate cancer. PSA also has been valuable in the accelerated development of therapies for prostate cancer; however, it must be used cautiously for this purpose, because it may not reflect the most relevant clone. In addition, some agents may directly affect PSA release independent of their antitumor activity. Most importantly, before PSA is adopted as a surrogate end point in clinical trials in prostate cancer, it must be prospectively validated. Future studies must focus on the development of prospective serologic tumor markers that can predict virulence of disease and to reflect androgen-independent progression. PMID- 10361552 TI - Prostate-specific antigen and other markers of therapeutic response. AB - Several new agents and combinations demonstrate significant activity in the treatment of patients with hormone refractory prostate cancer. Prostate-specific antigen (PSA) is being used increasingly as the key marker of a therapeutic response in trials of new agents. This article reviews data that support this marker as a surrogate endpoint, and it discusses the issues around the appropriateness of PSA as an intermediate marker with evolving noncytotoxic agents. Other biomarkers of prostate cancer progression are not uniformly elevated in men with advanced disease; to date, they are of limited clinical use. This article also discusses the rationale and results of novel and alternative biomarkers of prostate cancer progression. PMID- 10361553 TI - Prognostic indicators in hormone refractory prostate cancer. AB - Prognostic factors in hormone refractory prostate cancer currently are of limited use to clinicians. Although studies have identified several factors that predict for poor survival in patients, most are either retrospective, or nonrandomized. Therefore, large prospective, randomized trials are needed to validate the significance of these factors. In addition, these indicators are largely descriptive of the patients' condition or the extent of disease. As more treatment options are developed for these patients, functionally relevant and prognostic molecular markers are needed to direct their care. PMID- 10361554 TI - Secondary hormonal manipulations in hormone refractory prostate cancer. AB - Hormone refractory prostate cancer is clinically heterogeneous, and many patients retain sensitivity to subsequent hormonal manipulations, even after combined androgen blockage. Antiandrogen withdrawal is a mandatory first step. Subsequent treatment with an alternate antiandrogen, adrenal androgen inhibitor (such as ketoconazole), or glucocorticoid may provide both subjective and objective clinical benefit in up to 65% of patients. PMID- 10361555 TI - Chemotherapy for hormone refractory prostate cancer. AB - New combinations have been developed that show significant activity in therapy for hormone refractory prostate cancer. Several of these are designed to address specific cellular targets unique to prostate cancer. To date, the major benefits of these therapies have been palliative in nature, resulting in an improvement in quality of life, particularly with the combination of mitoxantrone and prednisone. None of these agents or regimens have been shown to affect survival significantly, and none can be considered to be standard therapy for this disease. Nonetheless, the success of these regimens in inducing response has challenged the skepticism concerning the appropriateness of chemotherapy for patients with advanced prostate cancer. The ability to slow and even reverse the growth of far advanced disease raises the possibility that the application of these regimens earlier in the course of the disease will have a more significant impact on the morbidity and, in the long run, on the mortality of prostate cancer. It is hoped that the enrollment of patients into properly designed clinical trials of new agents and combinations will result in the development of therapy with proven efficacy in the near future. PMID- 10361556 TI - Oral chemotherapy for hormone refractory prostate cancer. The University of Michigan experience. AB - The use of oral chemotherapy for the treatment of malignant disease is expanding. The authors' experience with oral chemotherapy for hormone-refractory prostate cancer continues to grow. These therapies are well-tolerated and effective. Already, these regimens are being improved by hybridizing them with intravenous agents such as paclitaxel. Also, oral novel agents are being tested that may offer new options for the treatment of hormone-refractory prostate cancer. PMID- 10361557 TI - Differentiating agents and nontoxic therapies. AB - The discovery of the oncogene and the mechanism by which these genetic changes create malignant transformation has provided new opportunities for drug development. Suramin is the first drug shown to exert its anticancer activity by blocking autocrine loops involved in malignant transformation. Phenylacetate and related aromatic fatty acids are potent inducers of differentiation in normal and malignant cells. Arachidonate, a fatty acid, plays a role in prostate cancer survival, growth, invasiveness, and immunosuppression. The actions of arachidonic acid can be moderated by diet or blocked by pharmacologic agents. Other agents that promise low toxicity include vitamin D and its analogs, genistein and related isoflavones, green tea polyphonols, and retinoic acid analogs. PMID- 10361558 TI - Gene therapy for prostate cancer. New perspectives on an old problem. AB - Recent advances in molecular biology have made the prospect of gene therapy for prostate cancer a reality. A wide variety of genetic strategies, vector designs, and delivery modalities are currently in use. This article examines the state of the art prostate cancer gene therapy and details the various options available to clinicians. PMID- 10361559 TI - Vaccine therapy for prostate cancer. AB - Vaccine therapy may provide an alternative for prostate cancer patients whose disease no longer responds to hormone therapy. Administration of dendritic cells pulsed with prostate-specific membrane antigen (PSMA) induces cellular immune responses against the tumor with virtually no adverse effects. About 30% of the evaluable patients were identified as partial responders, based on the National Prostate Cancer Project (NPCP) criteria. In addition, there was a 50% decrease of serum prostate-specific antigen or resolution of previously measurable lesions on imaging. Dendritic cell vaccine therapy may have a synergistic effect, when combined with other therapies. PMID- 10361560 TI - Supportive care, pain management, and quality of life in advanced prostate cancer. AB - Despite achievements in the area of providing care for patients with advanced prostate cancer, ample work remains. Additional research is needed regarding the control of pain from bone metastases and the management of fatigue and urinary symptoms. Investigators have only begun to explore the area of quality of life research in patients with prostate cancer. Other issues not addressed in this article that are significant to the care of these patients include caregiver burden and end-of-life care. These areas significantly affect quality of life. The supportive care, pain management, and quality of life issues discussed herein present many challenges to health care providers. Close attention to what patients tell us about their care will make the challenge more attainable and the caregiving more satisfying. PMID- 10361561 TI - Local and systemic radiation for palliation of metastatic disease. AB - Many radiotherapeutic treatment options are available for the palliation of patients with metastatic prostate cancer. These include local field radiotherapy to symptomatic sites of metastasis and the use of radioisotope therapy either alone or in combination with local field radiotherapy. To date, the majority of patients treated with radioisotope therapy have been treated with 89Sr. Other agents, such as 153Sm-EDTMP are available now, also. Combined radioisotope therapy, cytotoxic chemotherapy, and biphosphonates hold great promise. PMID- 10361562 TI - Nutrition and prostate cancer. AB - Scientific evidence suggests that differences in the diet may, in large part, account for the variability of prostate cancer rates around the world. Epidemiologic studies and animal experiments have yielded compelling results to warrant clinical intervention studies on nutrition from scientists who work on the prevention and treatment of prostate cancer. This article reviews the most recent evidence as to possible mechanisms of action of various dietary constituents, and explores evidence of various nutritional strategies for the prevention of prostate cancer progression. PMID- 10361563 TI - Alternative therapies for advanced prostate cancer. What should I tell my patients? AB - Alternative medicine use in the general population has continued to dramatically increase in this decade. The research supporting the use of acupuncture treatment in other areas of medicine is compelling. In addition, herbal therapies, such as PC-SPES and St. John's Wort, may play important roles in this disease. Clinicians dealing with advanced prostate cancer need to be introduced to some of these newer treatments so they can be discussed objectively with patients. PMID- 10361564 TI - A transvaginal sling procedure with bone anchor fixation. AB - Urethral sling procedures have become increasingly popular in the treatment of female stress incontinence. Until recently, the use of this procedure in the urologic community has been limited by technical difficulties and complications (e.g., urinary retention, urethral injury, urge frequency). Many modifications of the original sling procedure recently have been described to decrease surgical morbidity. This article describes a minimally invasive sling procedure performed completely through the vagina, with the aid of bone anchor fixation. The potential advantages of this operation include a rapid return to full activity and normal voiding. It should be noted that neither the long-term safety nor the efficacy has been established. PMID- 10361565 TI - National Health Service funding for complementary medicine research: the NHS acupuncture trials. PMID- 10361566 TI - Acupuncture for migraine and headache in primary care: a protocol for a pragmatic, randomized trial. AB - This paper presents the protocol for a randomized trial of acupuncture for migraine and headache. SUBJECTS: Four hundred patients with migraine or headache will be recruited from GP practices. INCLUSION CRITERIA: Eighteen to 65 years old, contractable by telephone, onset at least 1 year prior at age less than 50, two headaches per month in the previous 6 months, adequate data completion and headache severity during pre-randomization baseline. EXCLUSION CRITERIA: Pregnancy or malignancy, cluster headache, serious pathological aetiology, cranial neuralgia, acupuncture treatment in the past year. DESIGN: Following a 4 week baseline, patients will be allocated to acupuncture or control by minimized randomization. Up to 12 acupuncture treatments will be provided by advanced members of the Acupuncture Association of Chartered Physiotherapists. The type of acupuncture given will be recorded. STUDY MEASURES: Outcome will be assessed by headache diary, medication diary and SF36 at 3 months and 1 year. Resource use and days off sick will be assessed by quarterly questionnaire. Adverse events will be monitored by self-report. The primary outcome measure will be the change in mean daily headache score between baseline and the 1 year follow-up. An economic evaluation will also be undertaken. PMID- 10361567 TI - Reviewer bias against the unconventional? A randomized double-blind study of peer review. AB - OBJECTIVE: To test the hypothesis that there is a reviewer bias against publication of a test of an unconventional drug. DESIGN: Randomized, controlled, double-blind study of peer review. PARTICIPANTS: Convenience sample of 291 medical doctors from a wide variety of specialties drawn from a list of conference participants. METHODS: Reviewers were randomly assigned to receive one of two versions of a manuscript. Version M related to an in-vitro experiment on a mainstream drug (Metoprolol). The otherwise identical version V used a highly unconventional drug (beef spleen cell extract) for the same experiment. Reviewers were asked to complete a standardised evaluation sheet including visual analogue scales (VASs) on a set of predefined quality criteria. All participants were debriefed after completion of the study. RESULTS: The response rate was 61%. There were no significant differences in VAS ratings between the two versions of the manuscript. Ratings covered the entire range of the VASs. CONCLUSION: In the present setting, there was no evidence for a reviewer-bias against testing an unconventional drug. The low inter-rater reliability, however, suggested inadequate validity of peer review. PMID- 10361569 TI - 'Complementary' or 'alternative'? It makes a difference in cancer care. PMID- 10361568 TI - Efficacy of a potentized homoeopathic drug (Arsenicum Album-30) in reducing toxic effects produced by arsenic trioxide in mice: II. On alterations in body weight, tissue weight and total protein. AB - OBJECTIVE: To study the alterations in body weight, tissue weight and total protein in mice, caused by a single sublethal injection of arsenic trioxide and to investigate whether treatment by microdoses of arsenic has any antidotal effect. METHODS: For each fixation interval, altogether 36 individuals of Swiss albino mice, Mus musculus, were used, 27 were injected with As2O3 in a single sub lethal dose (@1.0 mg/kg body weight) and were divided into three batches. One batch was fed with diluted potentized alcohol (Alcohol control), one batch was fed with potentized homoeopathic drug Ars.Alb-30 (Active treatment), while the remaining one neither fed with potenized alcohol nor with the potentized homoeopathic drug (As-intoxicated control). The remaining batch of nine mice were injected with normal saline which served as negative control (Saline control). The mean body weights before and after injections and weights of different tissues like liver, kidney, spleen and testis were recorded at seven fixation intervals, 12 hours, 24 hours, 48 hours, 7 days, 21 days, 30 days, and 90 days. RESULTS: In arsenic treated mice orally administered with the homoeopathic drug statistically significant increases were noted in the weights of individual tissue weight, protein content as well as the mean body weight as compared to their respective controls. CONCLUSIONS: Arsenicum album can be considered as an antidote to arsenic poisoning. PMID- 10361570 TI - Familiarizing medical students with complementary and alternative medicine: encouraging new attitudes and ideas. PMID- 10361571 TI - Improving research quality and its use in service development. PMID- 10361572 TI - The function of small GTPases in signaling by immune recognition and other leukocyte receptors. PMID- 10361573 TI - Function of the CD3 subunits of the pre-TCR and TCR complexes during T cell development. PMID- 10361574 TI - Inhibitory pathways triggered by ITIM-containing receptors. PMID- 10361575 TI - ATM in lymphoid development and tumorigenesis. PMID- 10361576 TI - Comparison of intact antibody structures and the implications for effector function. PMID- 10361577 TI - Lymphocyte trafficking and regional immunity. PMID- 10361578 TI - Dendritic cells. PMID- 10361579 TI - Integrins in the immune system. PMID- 10361580 TI - Women's Health: Occupation, Cancer, and Reproduction. A conference overview. PMID- 10361581 TI - Occupational cancer among women: research status and methodologic considerations. AB - Occupational causes of cancer have not been well-evaluated among women. An increase in the number of women in the work force in jobs with potentially hazardous exposures during the past few decades raises the question as to whether there is a need to enhance our efforts in this area. The inability to evaluate occupational causes of female gynecologic tumors in studies of men, plus the potential for variation in outcome responses between men and women because of gender-based exposure and susceptibility differences, underscore the need for investigations specifically focused on women. Investigations of occupational exposures and cancer risk among women may require design considerations that differ somewhat from studies of men. Issues to consider include the impact of studying outcomes with high survival (e.g., breast cancer), gender-specific exposure patterns and toxicokinetic processing of some chemicals, special limitations in the use of the general population as the referent, and the need to control for established risk factors for gynecologic tumors. PMID- 10361582 TI - Women's reproductive health: some recent developments in occupational epidemiology. AB - Epidemiological research on occupational hazards and reproductive health is an expanding and strongly developing area. This article focuses on some recent areas of occupational reproductive epidemiology that are or seem to be important for the future. Interest in the research on fertility has increased during the past decade, and time to pregnancy has proved to be a useful measure of fertility. The research on menstrual function or early fetal loss is still limited, and further research is desirable. It is important to chart the advantages and disadvantages of various methods for measuring these outcomes. Recently developed methods of exposure assessment provide new possibilities to improve the validity of exposure data. Biological exposure markers can also provide useful dosimeters for reproductive studies. Research on the reproductive effects of job stress and individual susceptibility to reproductive toxicants is also gaining in importance. PMID- 10361583 TI - Measuring aspects of women's life and work for the study of variations in health. AB - BACKGROUND: Studies of occupation and health commonly examine only men. This paper draws on reviews of inequalities in health carried out by the Office for National Statistics in London which in recent years have focused more on women. METHODS: Many sources of official statistics such as censuses, surveys, vital registration and health service records are used to explore different ways of measuring and monitoring economic activity, education, socio-economic, and family status and their impact on fertility and health among women. RESULTS: Taking a life cycle approach we first look at fertility and family formation, the time around childbirth, age 15-49, and finally 50+. Some relevant health issues are used to illustrate how we use available data to describe and monitor inequalities in health. CONCLUSIONS: In Britain, there is a wealth of information and linked data sources which enable us to analyze patterns and trends in fertility, morbidity, and mortality. Nevertheless, none of the traditional data sources or methods of analysis are suitable for all purposes nor for keeping up with changes in society. Sources of official statistics and ways of linking and analyzing need then to be continuously developed. PMID- 10361584 TI - Women's occupational health in globalization and development. AB - BACKGROUND: The article describes the current process of globalization and its implications for development generally and for women, their work, and health. METHODS: The article outlines positive impacts in terms of enhancing employment opportunities in nontraditional spheres, and negative impacts in the growth of poor quality, insecure jobs with weakened social support systems. The case study of women's work within export processing zones is used to explore these conditions and their health impacts. RESULTS: The case study and other evidence provides a profile of work-related health that arises from a mix of patterns of employment, work processes, living conditions, and reproductive rights. CONCLUSIONS: The observed problems are poorly recognized, poorly studied for their combined causes and outcomes, and poorly regulated. The article explores and reviews how the patterns of female employment affect women's ability to collectively confront the causes of ill health and the challenges to improving women's occupational health in these conditions. PMID- 10361585 TI - Breast cancer risk among relatively young women employed in solvent-using industries. AB - BACKGROUND: Breast cancer is the most common tumor among women, and the causes remain almost unknown apart from changes in the reproductive pattern. Based on experimental evidence, some organic solvents may have carcinogenic properties to the female breast. METHODS: We used a comprehensive national data linkage to examine the adjusted breast cancer risk among relatively young (20-55 years) Danish women employed in industries with extensive use of organic solvents (i.e., the metal product, wood and furniture, printing, chemical, and textile and clothing industries). Relative risks (OR) were estimated from a matched case control study on 7,802 women with breast cancer (1970-1989). Potential exposure to organic solvents was accessed from the duration of employment within the selected industries and reconstructed from the files of a nationwide compulsory pension fund. Socioeconomic status and the individual reproductive pattern were obtained from the central person registry. RESULTS: The adjusted OR for breast cancer after 15 years latency was increased in each of the selected industrial groups (from 1.4 to 2.4). For the entire group with over 10 years of employment, the OR was significantly elevated (twofold). CONCLUSIONS: This study supports the observation that long-term occupational exposure to organic solvents may play a role in breast cancer risk. However, some residual confounding may exist, and further studies are required to identify specific carcinogenic organic solvents. PMID- 10361586 TI - Breast cancer and occupational exposures in women in Finland. AB - BACKGROUND: The etiology of breast cancer is not fully understood. Environmental and occupational exposures may contribute to breast cancer risk. METHODS: We linked 324 job titles from the 1970 census of 892,591 Finnish women with incidence of breast cancer (23,638 cases) during 1971-1995. We converted job titles to 31 chemical and two ergonomic agents through a measurement-based, period-specific, national job-exposure matrix. Poisson regression models were fit to the data, with adjustment for birth cohort, follow-up period, socioeconomic status, mean number of children, mean age at first delivery, and turnover rate. RESULTS: For premenopausal breast cancer, medium/high level of occupational exposure to ionizing radiation was associated with a standardized incidence ratio (SIR) of 1.3 (95% confidence interval (CI) 0.7-2.5; trend P = 0.03). For postmenopausal breast cancer, we found on SIR of 1.2 (1.1-1.3) for low level and 1.4 (1.1-1.8) for medium/high level of ionizing radiation (trend P = 0.001); and an SIR 1.3 (1.1-1.7) for medium/high levels of both asbestos and man-made vitreous fibers. Aromatic hydrocarbon solvents showed a significant trend for a modest excess of postmenopausal breast cancer. CONCLUSIONS: Our study indicates that occupational exposure to ionizing radiation may be associated with an increased risk of female breast cancer. High-quality studies on environmental and occupational etiology of breast cancer are needed for further elucidation of risk factors. PMID- 10361587 TI - Gender differences in risk of renal cell carcinoma and occupational exposures to chlorinated aliphatic hydrocarbons. AB - BACKGROUND: Organic solvents have been associated with renal cell cancer; however, the risk by gender and type of solvents is nuclear. METHODS: We evaluated the risk of renal cell carcinoma among men and women exposed to all organic solvents-combined, all chlorinated aliphatic hydrocarbons (CAHC) combined, and nine individual CAHC using a priori job exposure matrices developed by NCI in a population-based case-control study in Minnesota, U.S. We interviewed 438 renal cell cancer cases (273 men and 165 women) and 687 controls (462 men and 225 women). RESULTS: Overall, 34% of male cases and 21% of female cases were exposed to organic solvents in general. The risk of renal cell carcinoma was significantly elevated among women exposed to all organic solvents combined (OR = 2.3; 95% CI = 1.3-4.2), to CAHC combined (OR = 2.1; 95% CI = 1.1-3.9), and to trichloroethylene (TCE) (OR = 2.0; 95% CI = 1.0-4.0). Among men, no significant excess risk was observed among men exposed to any of these nine individual CAHCs, all CAHCs-combined, or all organic solvents-combined. DISCUSSION: These observed gender differences in risk of renal cell carcinoma in relation to exposure to organic solvents may be explained by chance based on small numbers, or by the differences in body fat content, metabolic activity, the rate of elimination of xenobiotics from the body, or by differences in the level of exposure between men and women, even though they have the same job title. PMID- 10361588 TI - Occupational, environmental, and life-style factors associated with the risk of hematolymphopoietic malignancies in women. AB - BACKGROUND: The etiology of lymphomas, leukemias, and multiple myeloma is still largely unknown. The known risk factors (ionizing radiation, solvent exposure, pesticide exposure, immunosuppression) explain only a small proportion of the cases that occur. METHODS: We conducted a multicenter population-based case control study on hematolymphopoietic malignancies in Italy and interviewed 2,011 women (1,183 cases and 828 controls). RESULTS: There was a suggestion of a positive association between smoking and the risk of non-Hodgkin's lymphoma + chronic lymphocytic leukemia. A slight increased risk of leukemias was observed among women using permanent hair dye. Housewives were at increased risk for leukemia and multiple myeloma. The risk of non-Hodgkin's lymphomas + chronic lymphocytic leukemia, leukemias, multiple myeloma, and Hodgkin's disease increased among women employed as hairdressers and textile workers. Teachers were at increased risk for non-Hodgkin's lymphomas + chronic lymphocytic leukemia, leukemias, and Hodgkin's disease. CONCLUSIONS: These results confirm previous associations and may provide additional clues to some determinants of hematolymphopoietic malignancies in women. PMID- 10361589 TI - Occupational risk factors for cancer of the central nervous system (CNS) among US women. AB - BACKGROUND: In a recent report, we found an elevated risk of cancer of the central nervous system (CNS) in several occupations and industries, and a modest association with exposure to solvents and to contact with the public. METHODS: To further explore the occupational risk of CNS cancer among women, we extended the analysis of the previous death certificate-based case-control study, including 12,980 female cases (ICD-9 codes 191 and 192) in 24 US states in 1984-1992 and 51,920 female controls who died from diseases other than malignancies and neurological disorders. We applied newly designed job-exposure matrices for 11 occupational hazards, previously reported as brain cancer risk factors, to the occupation and industry codes in the death certificates. We also conducted a separate analysis of 161 meningioma cases (ICD-9 codes 192.1 and 192.3), a tumor more frequent among women, particularly in the postmenopausal age group. RESULTS: Overall, CNS cancer risk showed a 20-30% increase among women exposed to electromagnetic fields (EMF), methylene chloride, insecticides and fungicides, and contact with the public. Risk for meningioma was elevated among women exposed to lead (OR = 1.9; 95% CI 1.0, 3.9). CNS cancer did not show a clear pattern of risk increase by probability and intensity of exposure to any of the explored risk factors. Cross-classification by probability and intensity of exposure did not reveal any significant trend. Cases were too few to explore trends of meningioma by probability and intensity of exposure to lead. CONCLUSIONS: We did not find evidence of a strong contribution of 11 occupational hazards to the etiology of CNS cancer. However, limitations of the occupational information might have reduced our ability to detect clear patterns of risk. PMID- 10361590 TI - Smoking as a confounder in case-control studies of occupational bladder cancer in women. AB - BACKGROUND: In studies in men, risk estimates on occupation and bladder cancer are distorted by about 10% when not adjusting for smoking. We examined the degree to which occupational risk estimates for bladder cancer in women are confounded by smoking, and the degree of residual confounding by inadequate control of this effect. METHODS: Primary data of 11 case-control studies on occupation and bladder cancer from Denmark, France, Germany, Greece, Italy, and Spain were pooled. Information for smoking and lifetime occupational history for 700 female cases and 2,425 female controls ages 30-79 was abstracted and recoded. Logistic regression was used to calculate odds ratios (OR) by occupation, applying five models which differed in their degree of adjustment for smoking. RESULTS: In major occupational groups, risk estimates were distorted by less than 10% when not adjusting for smoking. A statistically significant excess risk for bladder cancer was found in 13 specific occupations and industries. In most occupations, adjustment for smoking led the ORs towards the null value, but all statistically significant associations were maintained after adjustment. In three occupations (lathe operators, field crop workers, and wood manufacturers), a statistically significant excess risk was masked when not adjusting for smoking. In six occupations, estimates were distorted by more than 10% (-22% up to +40%). In occupations where smoking acted as a positive confounder, the proportion of confounding removed using a dichotomous smoking variable (ever/never) was around 60%. In one occupation (buyers), controlling for smoking status (ever, never) led to over-adjustment, because the percentage of smokers was high but the quantity smoked was low. PMID- 10361591 TI - Ovarian cancer and occupational exposures in Finland. AB - BACKGROUND: No single occupational or environmental agent has been established as causing ovarian cancer, existing studies often being based on ecologic or proportional mortality data in which potential confounders related to reproductive history have not been taken into account. METHODS: This study linked 324 job titles of occupationally active Finnish women (n = 892,591) at 1970 census with incidence of ovarian cancer (Finnish Cancer Registry, 5,072 cases) during 1971-1995 (over 15 million person-years). The job titles were converted into indicators of exposure to 33 agents, using a national job-exposure matrix based on measurements and surveys (FINJEM). Poisson regression analyses were performed with stratification by birth cohort, follow-up period, and socioeconomic status, and adjusted for mean number of children, mean age at first delivery, and turnover rate for each job title. RESULTS: We found indications of elevated risks for aromatic hydrocarbon solvents (standardized incidence ratio 1.3 (95% CI 1.0-1.7), leather dust (1.4; 0.7-2.7), man-made vitreous fibers (1.3; 0.9-1.8), and high levels of asbestos (1.3; 0.9-1.8), and diesel (1.7; 0.7-4.1), and gasoline (1.5; 1.0-2.0) engine exhausts). Previously reported findings for hairdressers and women in the printing industry were supported in our data, but not for women in dry cleaning jobs. CONCLUSIONS: Given the various drawbacks in linkage studies and job-exposure matrices, the excesses found in this study need confirmation in individual-level studies. PMID- 10361592 TI - Occupational risk factors for lung cancer in women: results of a case-control study in Germany. AB - BACKGROUND: To investigate the association between lung cancer and occupational factors in women. METHODS: Six hundred eighty-six women with lung cancer and 712 controls matched for age and region were interviewed to gather occupational histories and information about other risk factors and confounders. Odds ratios (OR) and 95%-confidence intervals (CI) were calculated. RESULTS: There were 11 cases and 2 controls who reported occupational asbestos exposure. Significantly elevated risks (P < 0.05, smoking-adjusted), were observed in the following industries: chemical, oil (OR 3.7), pottery, glass (OR 2.5), engine, vehicle building (OR 2.2), paper, wood, print (OR 1.9), cleaning service, hairdressing, housekeeping, waste disposal (OR 1.5); and occupations: assemblers, unskilled metal workers (OR 2.5), stock clerks, etc. (OR 1.6), restaurant owners and hoteliers (OR 2.7), as well as waitresses and travel attendants (OR 2.6). CONCLUSIONS: The study provides evidence that both occupations previously observed as hazardous in males, and occupations of particular significance for women only, play a role in the risk of lung cancer in women. PMID- 10361593 TI - Sinonasal cancer, occupation, and tobacco smoking in European women and men. AB - BACKGROUND: In this analysis of European case-control studies on sinonasal cancer, we examined the risk for occupation and smoking, by gender and histological type. METHODS: The pooled data included 104 female and 451 male cases, and 241 female and 1,464 male controls. Lifetime smoking and occupational history were recoded following uniform criteria, and job-exposure matrices were applied for wood and leather dust. RESULTS: Wood dust exposure was associated with an excess risk in men (OR = 2.36, 95% CI 1.75-3.2) but not in women (OR = 1.17, 95% CI 0.31-4.47). Exposure to leather dust was associated with an excess risk in both genders. Both wood and leather dust were associated with adenocarcinomas rather than squamous cell carcinomas. Excess risks for smoking were higher for squamous cell carcinomas and higher in men than in women. CONCLUSIONS: In these European populations, occupation was associated with about 11% of all sinonasal cancers in women and 39% in men. This difference can, in part, be attributed to variation in exposure patterns between genders. PMID- 10361594 TI - Cancer incidence among women in the Norwegian pulp and paper industry. AB - BACKGROUND: The aim of the present study was to investigate cancer risk among women working in the Norwegian pulp and paper industry. The cohort included a total of 4,247 workers employed for at least one year between 1920 and 1993 (108,095 person-years), 85% of them as paper or administration workers. METHODS: The follow-up period for cancer was from 1953-1993. No data of exposure measurements were available. The analyses were based on comparisons of standard incidence ratios. The expected numbers of cancer cases were calculated using the five-year age-specific incidence rates for the entire female population. RESULTS: During the follow-up period, 380 new cases of cancer were observed vs. 322 expected (SIR = 1.2, 95% CI = 1.07-1.30). An excess risk of ovarian cancer was found (SIR = 1.5, 95% CI = 1.07-2.09). The SIR was highest among those younger than 55 years, and mostly among those working in paper departments. Short-term workers showed increased risk of lung and bladder cancer. CONCLUSIONS: Based on results from the present study, the increased risk of ovarian cancer is difficult to interpret, since existing knowledge of its etiology is limited. However, these women might have been exposed to various work-related agents such as talc, microbes, and different types of paper dust. PMID- 10361595 TI - Issues and findings in the evaluation of occupational risk among women high nickel alloys workers. AB - BACKGROUND: We present the mortality experience for a cohort of women (n = 2,877) from a large epidemiologic study of production and fabrication high nickel alloys workers (n = 31,165). All the plants were located within the United States and cohort eligibility required some work experience within the period of the late 1940s through the mid 1960s. METHODS: Vital status follow-up was through the end of 1988 and incorporated information from multiple sources. Cause-specific mortality was evaluated by comparing cohort mortality to the general United States female population and to local populations in geographic proximity to the plants. Relative risk estimates were determined for 62 cause of death categories using the Standardized Mortality Ratio (SMR) and were adjusted for age, race, gender, and calendar time by the indirect method using a modified life table technique. RESULTS: Relative risks for all causes (0.98), all cancers (0.90), lung cancer (1.34), and breast cancer (0.96) were nonsignificant when mortality was compared to the US female population. No relationship between mortality and length of time employed in the industry or work area was identified. DISCUSSION: Although there were some difficulties in tracing women due to name changes, comprehensive follow-up was obtained when using multiple sources of information. Our strategy resulted in resolving vital status for over 95% of the women, which is comparable to that of the male cohort. The type of jobs and work activities differed between genders. Females were employed predominantly in two work areas (allocated services, 87%, and grinding, 46%), whereas males were employed in several work areas (allocated services, 76%, grinding, 27%, hot working, 20%, and cold working, 17%). Considerable variation was noted among the study plants with respect to the percent of female production workers in the workforce. Generally, the patterns of relative risks derived for the total cohort and various subgroups are similar across the different comparison populations. Estimated elevated risks are usually lower when cohort mortality is compared to that of local populations. No increased risks were identified for any site-specific cancers or nonmalignant causes of death. PMID- 10361596 TI - Cancer mortality in women with probable exposure to silica: a death certificate study in 24 states of the U.S. AB - BACKGROUND: Silica exposure is known to cause an increased risk of pneumoconiosis and some types of cancers. Exposure to silica is becoming an increasingly common occupational hazard for women. Studies contradict each other on whether or not women suffer more occupational pneumoconiosis than men, but no studies have evaluated cancer risks among women exposed to silica. METHODS: Death certificate data on occupation and industry from 24 states in the U.S. between 1984 and 1993 were used to calculate proportional mortality ratios (PMRs) for workers exposed to silica. RESULTS: Over 20,000 deaths (4% of all deaths in persons with possible work-related silica-exposure) occurred among women. The PMR for pneumoconiosis among women working in occupations or industries with possible silica exposure was 13.6 (95% CI: 7.2-23.2), for men 3.8 (CI: 3.7-4.0). Both men and women had higher than expected PMRs for respiratory diseases, lung and esophageal cancers, and external causes of death. In the group with probable silica exposure (both occupation and industry associated with silica), women had elevated PMRs for thyroid cancer (PMR = 5.5), multiple myeloma (PMR = 1.3), digestive organ cancers (PMR = 1.2), whereas men had no increased PMRs for these cancers. Both genders had significantly decreased PMRs for breast cancer, cerebrovascular diseases, nervous system diseases, and brain and other central nervous system cancers. CONCLUSIONS: An in depth look at the types of silica exposures (specific work duties) and adjustment for confounders is warranted to determine the importance of these gender-specific excess mortalities associated with possible silica exposure. PMID- 10361597 TI - Cohort mortality study of women compensated for asbestosis in Italy. AB - BACKGROUND: The carcinogenic effect of asbestos is accepted for lung cancer and mesothelioma, while conflicting opinions exist for other cancer sites. The aim of the present investigation is to study cause-specific mortality of women compensated for asbestosis who had certainly been exposed to high levels of asbestos fibers. METHODS: The cause-specific mortality of all Italian women compensated for asbestosis and alive December 31, 1979, was investigated through October 30, 1997. In the total cohort, which included 631 subjects, 277 deaths occurred. Cause-specific SMRs (Standardized Mortality Ratio) were computed using the national rates for comparison. RESULTS: A significantly increased mortality for all diseases related to asbestos exposure was observed. Mortality for all causes, all neoplasms, lung cancer, uterine cancer, ovarian cancer, and non neoplastic respiratory diseases was significantly increased. Separate analyses for textile (n = 276) and asbestos-cement (n = 278) workers were performed. Women employed in the textile industry, mainly exposed to chrysotile, who are compensated at a younger age, showed higher SMRs for lung cancer and asbestosis. Women in the asbestos-cement industry, mainly exposed to crocidolite containing asbestos mixtures, experienced higher mortality for pleural malignancies. CONCLUSIONS: The role of asbestos exposure in the development of gastrointestinal and genital neoplasms is discussed. PMID- 10361598 TI - Cancer risk among female agricultural workers: a multi-center case-control study. AB - BACKGROUND: Cancer risk among women engaged in farming has been poorly investigated. This group of female workers is of particular interest, however, since they may experience exposure to several potential agricultural hazards. METHODS: A hospital-based case-control study was conducted in five Italian rural areas to examine the association between cancer and farming among women. The areas selected were located in three different regions (i.e., Piedmont, Tuscany, and Emilia-Romagna). The following cancer sites were selected for the study: stomach, colon, rectum, lung, skin melanoma, skin non-melanoma, breast, cervix and corpus uteri, ovary, bladder, kidney. Cases of non-Hodgkin's lymphoma were also included. Altogether, 1,044 newly diagnosed cases aged 20-75 years were ascertained from hospital records from March 1990 to September 1992, and for 945 of them detailed information was collected by a standard questionnaire. The analyses of data were performed comparing each case series to a reference group drawn from among the other sites. Unconditional logistic regression models were used in the statistical analyses. RESULTS: Statistically significant increased risks in association with farming were estimated for skin melanoma (OR 2.7, 95% CI 1.2-5.8) and bladder cancer (OR 2.7, 95% CI 1.2-6.1). Lung cancer was also found increased but not at a statistically significant level (OR 1.7, 95% CI 0.7 4.4). An OR lower than unity was observed for postmenopausal breast cancer (OR 0.4, 95% CI 0.3-0.7). CONCLUSIONS: The present study suggests that women in farming might experience increased risk of cancers, not usually found in excess among male farmers, as well as a protective effect for postmenopausal breast cancer. The role of different patterns of exposure or gender specific responses should be considered in further studies. PMID- 10361599 TI - Mortality among male licensed pesticide users and their wives. AB - BACKGROUND: We evaluated the mortality pattern of male licensed pesticide users and their wives in central Italy. METHODS: The cohort consisted of 2978 male farmers licensed for buying and handling toxic pesticides during the period 1971 1973 and 2586 farmers' wives. The Standardized Mortality Ratio (SMRs) and their 95% Confidence Intervals (95% CI) were computed on the basis of regional death rates. RESULTS: We found a lower than expected overall and cancer mortality. Non Hodgkin's lymphoma was increased among women (SMR = 2.29, 0.62-5.86) but not in male farmers (SMR = 0.90, 0.24-2.30), while both sexes were characterized by an increased risk of leukemia (men: SMR = 1.44, 0.69-2.64; women: SMR = 2.41, 1.04 4.76), mainly due to myeloid leukemia (men: SMR = 2.43, 0.98-5.00; women: SMR = 3.14, 1.02-7.33). CONCLUSIONS: Men and women tend to share the same mortality profile. The statistically significant increase of leukemia with a threefold increased risk of the myeloid subtype only among women suggests that different pattern of exposure or biological differences between genders should be considered in evaluating health risks in agricultural settings. PMID- 10361600 TI - Incidence of breast cancer in a Norwegian cohort of women with potential workplace exposure to 50 Hz magnetic fields. AB - BACKGROUND: The risk of breast cancer was investigated in a large dynamic population-based cohort of all 1.1 million economically active women in Norway with potential exposure to 50 Hz magnetic fields at the censuses of 1960, 1970, and 1980. METHODS: The follow-up period for the cohort was 1961-1992. For each woman, date of birth and census information on occupation and socioeconomic status were ascertained. These data were linked to the breast cancer morbidity information in the Cancer Registry of Norway. Exposure to magnetic fields was assessed a priori using two different approaches. In the first approach, hours per week in a potential magnetic field above background level (0.1 microT) were classified by an expert panel. In the second approach, measured magnetic fields from a separate study of men at work were allocated to the women's census job titles. In both approaches, exposure was cumulated over the years of employment (work hours and microT-years, respectively). RESULTS: The Poisson regression analysis showed a risk ratio (RR) of 1.14 (95% confidence interval (CI) = 1.10 1.19) in the highest exposure category compared to the lowest when using the first approach, and the corresponding RR was 1.08 (95% CI = 1.01-1.16) when using the second approach. For women younger than 50 years, RR was 1.20 (95% CI = 1.11 1.29) and 1.12 (95% CI = 0.98-1.28), respectively. CONCLUSIONS: The results give some support to the hypothesis that exposure to 50 Hz magnetic fields may increase the risk of breast cancer. However, since no direct information on exposure was available, no firm conclusions can be drawn. PMID- 10361601 TI - Cancer mortality in health and science technicians. AB - BACKGROUND: Nearly one million U.S. women are employed as health or science technicians with various chemical and biological exposures, but few studies have looked at their health outcomes. METHODS: Using 1984-1995 mortality data with coded occupation information, we calculated race- and age-adjusted proportionate cancer mortality ratios (PCMRs) and 95% confidence intervals for two age groups for black and white women with occupations of clinical laboratory (CLT), radiologic, and science technician. RESULTS: For CLTs, the PCMRs for breast cancer were borderline significantly elevated. The PCMRs for leukemia were significantly elevated, particularly for myeloid leukemia. Radiologic technicians had no significantly elevated PCMRs. Science technicians had significantly elevated PCMRs for non-Hodgkin's lymphoma and multiple myeloma in the younger age group. DISCUSSION: The elevated risks for lymphatic and hematopoietic neoplasms in CLTs and science technicians may be associated with occupational exposures. PMID- 10361602 TI - Cancer mortality among women employed in health care occupations in 24 U.S. states, 1984-1993. AB - BACKGROUND: Health care workers are potentially exposed to a number of carcinogens. Studies among women in this field have focused on white nurses; however, workers in many health care occupations share exposures experienced by nurses. METHODS: Cancer mortality was examined among female health care workers using death certificate data collected in 24 U.S. states from 1984 through 1993. Cancer mortality odds ratios (MORs) were calculated by race (white, black) and age group. RESULTS: White nurses had a 30% elevation of mortality due to liver cancer and myeloid leukemia. White registered nurses (RNs) had a small excess and white licensed practical nurses (LPNs) had a small deficit of mortality due to breast cancer. Ovarian cancer was in excess among RNs, but decreased among LPNs. Among black nurses, excesses of death due to kidney cancer (MOR = 1.7) and multiple myeloma (MOR = 1.3), and a significant 50% deficit in mortality due to cancer of the esophagus were found. Black RNs, but not LPNs, had an excess of breast cancer (MOR = 1.3; 95% CI = 1.0-1.5). Ovarian cancer was elevated by 30% in both RNs and LPNs. Excess deaths due to cancers of the breast, ovary, and uterus occurred among white physicians. Among black physicians, lung cancer was significantly elevated (MOR = 2.8). White pharmacists had significant excesses of breast (MOR = 1.5) and ovarian (MOR = 2.4) cancers, and myeloid leukemia (MOR = 2.0). White clinical laboratory technicians had excess deaths from several cancers. The greatest excess was for myeloid leukemia (MOR = 2.3; 95% CI = 1.5 3.4). Excesses among radiologic technologists included cancers of the lung, pancreas, breast, uterus, and ovary. CONCLUSION: Several findings reported here warrant further investigation. In particular, excesses of myeloid leukemia among nurses, pharmacists, and clinical laboratory technicians and liver cancer among nurses should be investigated in studies with data on occupational and other exposures. Patterns of mortality from breast and ovarian cancer found in this study must be evaluated further in studies with data on reproductive history. PMID- 10361603 TI - Cancer mortality among women in the Russian printing industry. AB - BACKGROUND: This study evaluates cancer mortality among women employed in two large printing plants in Moscow. METHODS: A total of 3,473 women who were actively employed as of December 31, 1978, with a minimum of 2 years employment were followed from 1 January 1979 to 31 December 1993. There were 47,791 person years observed, with only 51 women lost to follow-up (1.5%). Standardized mortality ratios (SMRs) were calculated using the population of Moscow to generate expected numbers. Analyses by job (compositors, press operators, and bookbinders), age hired, latency, and duration of employment were conducted. RESULTS: Among women employed in the two printing plants, there was a significant excess of esophageal cancer, based on seven deaths (expected = 2.7, SMR = 2.7, 95% CI = 1.1-5.4). Four of the seven esophageal cancer deaths occurred among bookbinders (expected = 1.0, SMR = 4.1, 95% CI = 1.1-10.4), all among workers hired before 1957 (expected = 0.6, SMR = 7.1, 95% CI = 1.9-18.3), the last year benzene was used in bookbinding. Ovarian cancer was also significantly elevated among bookbinders (12 observed, 4.2 expected, SMR = 2.9, 95% CI = 1.5-5.0), which, along with one death from mesothelioma of the abdomen, might be related to the use of asbestos-contaminated talc fillers in paper. Press operators had significantly elevated mortality from stomach cancer (observed = 9, expected = 4.1, SMR = 2.2, 95% CI = 1.0-4.2) and, based on two deaths each, melanoma and bladder cancer. CONCLUSIONS: Women in this printing industry cohort experienced excess mortality of cancer of the esophagus and stomach, with suggested increases of melanoma and bladder cancer. Further follow-up of this cohort, which would allow more in-depth analysis of rare cancer sites, latency, and duration of employment, is warranted. Gender comparisons within the cohort should also be conducted to clarify the role of occupational and lifestyle factors in the etiology of cancer among workers in the printing industry. PMID- 10361604 TI - Women's occupation and cancer: preliminary analysis of cancer registrations in England and Wales, 1971-1990. AB - BACKGROUND: In recent years increasing attention has been paid to examining the relation between women's health and their own occupation. The findings presented here relate to an analysis of 381,915 cancers in women registered in England and Wales over the 20-year period 1971-1990. METHODS: To explore the value of the data for epidemiological research, five sites (pleura, bladder, stomach, lung, and breast) and two occupations (agriculture and textile) were selected. Associations between cancer and occupation were evaluated using age and social class adjusted proportional registration ratios (PRRs) and 95% confidence intervals (CI). RESULTS: Pleural cancer risk was increased in a range of occupations with a potential for asbestos exposure (e.g., PRR 608, 95% CI 381-921 for craft and other production process workers), and the well-established link between bladder cancer and employment in the rubber industry was confirmed (PRR 303, 95% CI 188-464). Associations between "dusty" occupations and stomach cancer were noted (e.g., PRR 198, 95% CI 126-298 for ceramic decorators and finishers), and possible links with lung cancer and smoke in the work environment were highlighted (e.g., PRR 167, 95% CI 147-189 for publicans). As a group, agricultural workers were found to be at increased risk of non-Hodgkin's lymphoma (PRR 164, 95% CI 126-211), and textile workers at increased risk of pleural cancer (PRR 145, 95% CI 111-185). No evidence for an occupational aetiology for breast cancer was found within these data. CONCLUSIONS: Occupational information collected at cancer registration in the United Kingdom can be reliably used to identify groups that may be at increased risk of disease. PMID- 10361605 TI - Cancer mortality among women employed in fast-growing U.S. occupations. AB - Our study examined cancer mortality before the age of 65 for women employed in the fastest growing and/or traditionally female occupations. Analysis of mortality data from 28 U.S. states for 1984-1995 revealed elevated proportionate cancer mortality ratios (PCMRs). The highest PCMRs observed were thyroid cancer among health aides, lymphatic and multiple myeloma among computer programmers, and brain cancer among actresses and directresses. Some of the excess mortality occurred for occupations that have been previously cited. These included elevated breast and ovarian cancer among teachers, Hodgkin's disease among hairdressers and cosmetologists, and thyroid cancer among health aides and therapists. A few of the associations were new, i.e., had not been previously observed. These included cancer of the connective tissue and lymphatic system among computer programmers, ovarian cancer and leukemia among secretaries, and lymphatic cancer and multiple myeloma among child care workers. These findings should be further investigated with epidemiologic and environmental studies. PMID- 10361606 TI - Is there a healthy worker effect for cancer incidence among women in Sweden? AB - BACKGROUND: Our aim was to evaluate whether there is a healthy worker effect (HWE) for cancer incidence among women. HWE is a bias found in occupational studies that compare rates of disease among employed people to disease rates for the general population, which includes unemployed people (who may be less healthy than those who are employed). METHODS: Data from the 1960 and 1970 Swedish censuses were used to identify all 1,659,940 Swedish women who were employed in either year. They were followed during 1971-1989 through linkages to the national cancer and death registers. Standardized incidence ratios (SIRs) were computed comparing employed women to the 1,627,873 women who were not employed in either 1960 or 1970. RESULTS: For the 545,857 women employed in both 1960 and 1970, the SIR for all cancers combined was 1.05 (1.04-1.06). When specific cancer sites were analyzed separately, the highest cancer risks were for cancers of the lung and bladder (SIR = 1.2) and reproductive organs (breast, ovary, endometrium, and cervix SIR = 1.1). Overall cancer risks were highest among full-time workers, younger workers, urban workers, and workers with the highest socioeconomic status (based on the woman's job title). CONCLUSIONS: These results show no general HWE for cancer incidence among employed Swedish women. PMID- 10361607 TI - Heavy physical work during pregnancy--a risk factor for small-for-gestational-age babies in Poland. AB - BACKGROUND: Heavy physical work is still considered one of the most prevalent risk factors of negative pregnancy outcome. The present study was undertaken to evaluate the impact of heavy physical work during pregnancy, based on subjective assessment of workload, on the risk of delivering a small-for-gestational-age (SGA) baby. METHODS: Job characteristics were compiled for 1,064 working women from the Lodz region (8% population sample) based on questionnaire responses. Energy expenditure during daily work was estimated. SGA was diagnosed when the birth weight was below the 10th percentile of the standard curves for central Poland. The SGA group comprised 78 women who delivered SGA babies, while the non SGA group consisted of 986 women with eutrophic newborns. RESULTS: An excessive risk of SGA was found in the group reporting heavy physical effort at work. The physical effort perceived as heavy by pregnant women appeared to be a better predictor of SGA manifestation than the estimated energy expenditure at work. CONCLUSIONS: The information obtained from a pregnant worker should constitute an essential decisive factor for determining the time of work cessation during pregnancy. PMID- 10361608 TI - Reduced fertility among female wood workers exposed to formaldehyde. AB - BACKGROUND: The aim of the study was to investigate whether exposure to formaldehyde, organic solvents or other chemicals in the wood-processing industry affects the fertility of women. METHODS: For this purpose, a retrospective study on time to pregnancy was conducted among female wood workers who had given birth during 1985-1995. Data on pregnancy history, time to pregnancy, occupational exposures, and potential confounders were collected by a questionnaire; 64% (699/1,094) participated. The exposure assessment was conducted by an occupational hygienist. The data on time to pregnancy were analyzed with the discrete proportional hazards regression. RESULTS: Exposure to formaldehyde was significantly associated with delayed conception: adjusted fecundability density ratio, FDR, was 0.64 (95% CI 0.43-0.92). At high exposure if no gloves were used, the FDR was 0.51 (% CI 0.28-0.92). Exposure to phenols, dusts, wood dusts, or organic solvents was not related to the time to pregnancy. Additionally, an association was observed between exposure to formaldehyde and an increased risk of spontaneous abortion (concerning previous spontaneous abortions, reported by the women). Associations between exposure to formaldehyde or to organic solvents and endometriosis, and between exposure to organic solvents or to dusts and salpingo-oophoritis were also suggested. CONCLUSIONS: The study suggests that a woman's occupational exposure to formaldehyde has an adverse effect on fertility. PMID- 10361609 TI - Maternal occupational exposure to chemical substances and the risk of infants small-for-gestational-age. AB - BACKGROUND: The association between maternal occupational exposure to specific chemical substances (organic solvents, carbon tetrachloride, herbicides, chlorophenols, polychlorinated biphenyls, aromatic amines, lead and lead compounds, mercury and mercury compounds) and birth of small-for-gestational-age (SGA) infants was evaluated using data from a prospective cohort study of 3,946 pregnant women in West Germany from 1987 to 1988. METHODS: Occupational, medical, and psychosocial information was gathered through a questionnaire from pregnant women who were recruited between 15 and 28 gestational weeks. Exposure to chemical substances at the current workplace was assessed by a job-exposure matrix constructed by Pannett in 1985 and weighted for the number of working hours per week. Women not working at the time of the interview, women with multiple births, and women with stillbirths were excluded from analysis. Data were analyzed using dichotomous and polytomous logistic regression to control for age, smoking status, alcohol consumption, body mass index, and number of former births. RESULTS: The results of the dichotomous logistic regression analysis suggest that leather work might be associated with the birth of infants small-for gestational-age through exposure to chlorophenols (P = 0.02) and aromatic amines (P = 0.05). In the polytomous logistic regression analysis, only the association between exposure to mercury and growth retardation reached statistical significance (P = 0.02); however, the power of the study is limited. Further adjustment for income, shift work, and heavy physical work had no substantial effect on the results. CONCLUSIONS: These findings suggest that maternal exposure to specific chemicals at work may be a risk factor for the birth of SGA infants. PMID- 10361610 TI - Trends in myocardial perfusion single-photon emission technique. PMID- 10361612 TI - Common slow modulation of respiration, arterial blood pressure and cortical activity during sleep onset while napping. AB - Using healthy subjects, concomitant 30- to 60-s modulations of respiration, arterial blood pressure and EEG activity were found in 21 experiments about napping. Although mean arterial pressure (MAP) modulations above and below 1/30 Hz increased, in respiratory amplitude (RA) only the lower frequency components augmented significantly. This slow modulation of RA was found to be asymmetrical in time, the duration of RA decreasing parts in the modulation waves being 42% longer than the duration of RA increasing parts. The concomitance of the slow modulations in the different organ systems is accounted for by the influence of the common brainstem system (CBS), which regulates and integrates respiratory, cardiovascular and somatomotor systems and the adjustment of central nervous activity (vigilance). The described common, slow modulations outline the importance of dampening influences during sleep onset. They may provide an important tool for the investigation of the regulatory systems during sleep onset, as well as for investigations about sleep apnoea syndrome and Cheyne Stokes breathing. PMID- 10361611 TI - Prostaglandin inhibition causes an increase in reactive hyperaemia after ischaemic exercise in human forearm. AB - The hypothesis that prostaglandins contribute to the reactive hyperaemia after 5 min of ischaemia or 5 min of ischaemic exercise was investigated in six men by inhibiting prostaglandin production with ibuprofen (1800 mg) and indomethacin (225 mg) over 24 h before testing. Blood flow was measured continuously in the baseline and after ischaemia by combined pulsed and echo Doppler as the product of velocity and cross-sectional area. After 5 min of ischaemia, there were no differences in blood flow between placebo and the two drug conditions, except at 5 and 10 s when flow with indomethacin was greater than both placebo and ibuprofen. After 5 min of ischaemic exercise, blood flow was significantly greater as a consequence of increased vascular conductance in each of ibuprofen and indomethacin than placebo from 5 until 90 s of recovery. We conclude that prostaglandin inhibition had little or no effect on reactive hyperaemia after 5 min of circulatory occlusion alone, but that blood flow after ischaemic exercise was elevated due to increased vascular conductance when prostaglandin synthesis was inhibited. PMID- 10361613 TI - Non-invasive cardiac output assessment during moderate exercise: pulse contour compared with CO2 rebreathing. AB - The arterial pulse contour method called Modelflow 2.1 calculates stroke volume continuously, beat to beat, from the non-invasive blood pressure signal measured by Finapres or Portapres. Portapres is the portable version of Finapres. The purpose of this study was to compare cardiac output (CO) calculated using Modelflow 2.1 (COmf) with CO obtained by the CO2 rebreathing method (COre) during steady state at moderate exercise levels. Twelve subjects visited the laboratory twice and performed submaximal exercise on a bicycle ergometer at 20%, 40% and 60% of their individual peak power output (POpeak). The averaged correlation between COmf and COre gives an r-value of 0.69, whereas the slope and intercept of the regression line were 1.06 and 1.65 respectively. The averaged difference between COmf and COre was 2.27 +/- 3.91 min-1 (mean +/- standard deviation). However, the test-retest difference between COmf and COre was 2.5 +/- 3.1 and 0.5 +/- 1.31 min-1 respectively. These results suggest that Modelflow 2.1 is not an accurate method for estimating CO from non-invasive blood pressure data collected by Portapres during exercise at up to 60% of the individual POpeak corresponding with daily life activity. PMID- 10361614 TI - Influence of orthopaedic metal and high-density detection on body composition as assessed by dual-energy X-ray absorptiometry. AB - We examined the influence of orthopaedic material and computerized high-density detection (HDD) on analysis of bone mass and soft tissue composition performed by dual-energy X-ray absorptiometry (DXA). Measurements of total and regional bone area, bone mineral content (BMC), areal bone mineral density (BMD), lean tissue mass (LTM) and fat tissue mass (FTM) were made using a Norland XR-26 DXA scanner with dynamically changing samarium filtration. Twenty-one subjects who were free of metal implants were measured without and with a Biomet femoral prosthesis (titanium) placed on the proximal part of the femoral region. Twenty-one women with an endogenous prosthesis in the proximal femur were measured once. Analyses of tissue composition were performed without and with HDD using software provided by the manufacturer. Measurements were considerably affected by exogenous metal with overestimation of LTM and underestimation of FTM and bone area. BMC and BMD were over- or underestimated depending on the anatomical region. Enabling the HDD mode, values of bone area and tissue mass came closer to the expected values ( metal/-HDD) but were in general still significantly different from these. For the total body, the following significant changes were found after application of metal (+metal/-HDD vs. +metal/+HDD, mean values): bone area -19.8% vs. -6.9%, BMC +1.1% vs. -2.1%, BMD +26.5% vs. +4.7%, LTM +12.4% vs. +3.7%, FTM -15.8% vs. 7.0%. A similar pattern of change in tissue composition and bone area was found for the subregions of the body. Changes in tissue composition after HDD were similar in subjects with exogenous and endogenous metal, indicating that the experimental model was appropriate. In conclusion, measurements of tissue composition were substantially influenced by orthopaedic metal. HDD partly corrected for the artefacts induced by the metal. PMID- 10361615 TI - Interstitial lactate levels in human skin at rest and during an oral glucose load: a microdialysis study. AB - In vitro data have suggested that the skin is a significant lactate source. The purpose of the present study was to measure lactate and glucose concentrations in intact human skin in vivo using the microdialysis technique. Microdialysis fibres of 216 microns were inserted intradermally and perfused at a rate of 3 microliters min-1. In the first experimental protocol, dialysis fibres were calibrated by the method of no net flux in eight subjects. Skin lactate concentrations of 2.48 +/- 0.17 mmol l-1 were significantly greater than lactate concentrations of 0.84 +/- 0.15 mmol l-1 in venous plasma (P < 0.01). Glucose concentrations in skin and venous plasma were similar (5.49 +/- 0.18 vs. 5.26 +/- 0.24 mmol l-1). In the second experimental protocol, changes in lactate and glucose levels were studied in 10 subjects after an oral glucose tolerance test (OGTT). After the OGTT, plasma glucose and lactate levels increased by 54% and 39% to peak levels at 30 and 60 min respectively. In comparison, skin glucose and lactate increased by 41% and 18% at 60 and 90 min. No changes in skin blood flow were observed during the OGTT. The data suggest that resting skin is a significant lactate source with no significant lactate production during OGTT. The cellular source of lactate in the skin remains undetermined to date. PMID- 10361616 TI - Effects of breathing exercises on breathing patterns in obese and non-obese subjects. AB - Chest physiotherapy in connection with abdominal surgery includes different deep breathing exercises to prevent post-operative pulmonary complications. The therapy is effective in preventing pulmonary complications, especially in high risk patients such as obese persons. The mechanisms behind the effect is unclear, but part of the effect may be explained by the changes in breathing patterns. The aim of this study was therefore to describe and to analyse the breathing patterns in obese and non-obese subjects during three different breathing techniques frequently used in the treatment of post-operative patients. Twenty-one severely obese [body mass index (BMI) > 40] and 21 non-obese (BMI 19-25) subjects were studied. All persons denied having any lung disease and were non-smokers. The breathing techniques investigated were: deep breaths without any resistance (DB), positive expiratory pressure (PEP) with an airway resistance of approximately +15 cmH2O (1.5 kPa) during expiration, inspiratory resistance positive expiratory pressure (IR-PEP) with a pressure of approximately -10 cmH2O (-1.0 kPa) during inspiration. Expiratory resistance as for PEP. Volume against time was monitored while the subjects were sitting in a body plethysmograph. Variables for volume and flow during the breathing cycle were determined. Tidal volume and alveolar ventilation were highest during DB, and peak inspiratory volume was significantly higher than during PEP and IR-PEP in the group of obese subjects. The breathing cycles were prolonged in all techniques but were most prolonged in PEP and IR PEP. The functional residual capacity (FRC) was significantly lower during DB than during PEP and IR-PEP in the group of obese subjects. FRC as determined within 2 min of finishing each breathing technique was identical to before the breathing manoeuvres. PMID- 10361617 TI - Reproducibility of abnormal heart rate variability indices: the case of hypertensive sleep apnoea syndrome. AB - Before heart rate variability can be used as an investigational tool in the clinical setting, its reproducibility must be known. We studied heart rate variability four times during 44 weeks in 15 hypertensive patients with sleep apnoea syndrome. Time and frequency domain analytical approaches were used during the spontaneous and controlled breathing tests, orthostatic manoeuvre and the cold pressor test. Alterations in resting heart rate were taken into account using the coefficient of component variance. In general, the response of heart rate variability was abnormal and variability was reduced in the hypertensive patients with sleep apnoea syndrome compared with reference data. Time domain measures of heart rate variability demonstrated generally better reproducibility over four recordings than frequency domain measures in these hypertensive patients with sleep apnoea syndrome. On the other hand, the reproducibility of frequency domain measures depended on the specific conditions: during orthostatic manoeuvre and cold pressor test the best reproducibility was found using normalized units. In the reference data set, there were no significant differences between the two heart rate variability recordings during any of the autonomic nervous function tests. In this follow-up study we found that simple procedures such as the controlled breathing test show good reproducibility. More complex tests such as the orthostatic manoeuvre require special attention in order to obtain acceptable reproducibility of heart rate variability measurements. Quantitatively minor changes in the variability indices when the overall variability is reduced exert major effects on the results. Therefore we suggest that reproducibility data obtained in healthy volunteers are not valid for the interpretation of data in patients with damaged cardiovascular autonomic control. PMID- 10361618 TI - The effect of breathing frequency on deposition of drug aerosol using an inhalation-synchronized dosimeter in healthy adults. AB - The deposition of inhaled drug aerosol between the tongue, the upper and lower respiratory tract, the lungs and the gastrointestinal tract (GI tract) in 11 healthy adults was studied by using a nebulizer with an inhalation-synchronized dosimeter. The effect of breathing frequency on deposition was studied using radioaerosol (mixture of salbutamol and technetium bound to diethylenetriamine pentacetate, [99mTc]DTPA) and a gamma-camera. In healthy subjects who were breathing at their own frequency (16 +/- 5 breaths min-1, mean +/- SD), the proportion of inhaled aerosol deposited in the lungs was 48 +/- 14 (mean percentage +/- SD). The proportion deposited in the upper airway tract and the GI tract was 19 +/- 13 and 25 +/- 9 respectively, and the remainder was deposited on the tongue (6 +/- 4) and in the lower airway tract (3 +/- 2). Guided, slower breathing frequency (11 +/- 5 breaths min-1) changed the deposition remarkably. The proportion of the pulmonary deposition of the inhaled dose increased significantly (P < 0.004) to 60 +/- 17, and the proportion of the upper airway tract deposition decreased significantly (P < 0.005) by half of the initial deposition. We conclude that a slow controlled breathing frequency is an important factor if we want to increase the drug deposition in the lungs. It is also essential in decreasing the variation in the deposition of the lungs. PMID- 10361619 TI - Both leg blood flow and the femoral blood flow pattern are related to left ventricular mass in elderly men. AB - Left ventricular mass (LVM) was significantly related to both the leg blood flow (r = -0.31, P < 0.05) and a Doppler-derived index of femoral arterial stiffness (r = 0.35, P < 0.05) in a sample of elderly men. Together with blood pressure, these two characteristics of peripheral blood flow explained 42% of the variation in LVM. PMID- 10361620 TI - Interaction between broad-spectrum antibiotics and the combined oral contraceptive pill. A literature review. AB - There is considerable variation in opinion about the importance of drug interactions between the combined oral contraceptive pill (COCP) and broad spectrum antibiotics. Clinical practice varies widely, especially between doctors in Europe and those in the US. Rifampicin and griseofulvin induce hepatic enzymes and do appear to have a genuine interaction with the COCP, leading to reduced efficacy. The situation with the broad-spectrum antibiotics is less clear. There are relatively few prospective studies of the pharmacokinetics of concurrent COCP and antibiotic use and few, if any, demonstrate a convincing basis for any reduced contraceptive efficacy. There is evidence, however, that variable contraceptive steroid handling could make some women, at some times, more susceptible to COCP failure. Given the serious consequences of unwanted pregnancy, the cautious approach of using additional or alternative contraception during short courses of broad-spectrum antibiotics and the initial weeks of long term antibiotic administration may be justified to safeguard the few unidentifiable women who may be at risk. Conflicting opinion and advice is potentially confusing to both professionals and patients, and instructions for additional precautions during and after concurrent COCP and antibiotic use are complicated. Many women are ignorant of, or confused about, the circumstances that can cause OC to fail. Health professionals who prescribe the COCP must continue to strive to educate women about the mode of action and about the times when there is the greatest danger of failure. Professionals who feel that concurrent antibiotic use represents a real threat to contraceptive efficacy of the COCP should be prepared to present the advice for additional contraceptive precautions in a simple and consistent way, backed up with written information and reinforced at regular intervals. PMID- 10361621 TI - The risk of venous thromboembolism in users of postcoital contraceptive pills. AB - Postcoital contraceptive pills (PCP) have recently been approved for use as emergency contraception in the United States. The objective of this study was to assess the risk of idiopathic venous thromboembolism (VTE) in relation to exposure to PCP, and to better quantify the risk of idiopathic VTE associated with current oral contraceptive (OC) use and pregnancy. A population-based cohort study with a nested case-control analysis was conducted using women from the General Practice Research Database. There were no women with an outcome of idiopathic VTE with current exposure to PCP. The incidence rates for various exposures were 3.0/100,000 person-years for the unexposed, 5.3/100,000 person years for second generation OC, 10.7/100,000 person-years for third generation OC, and 15.5/100,000 person-years in pregnant (or postpartum) women. The relative risk estimates were 1.7 (95% CI 0.3-10.5) for second generation OC, 4.4 (95% CI 1.0-18.7) for third generation OC, and 6.3 (95% CI 1.2-33.5) for pregnancy. Short term use of PCP is not associated with a substantially increased risk for developing VTE. PMID- 10361622 TI - Contraceptive status and sexual function of climacteric Chinese women. AB - The objective of the present survey was to assess the contraceptive status and sexual function of climacteric Chinese women. One cross-sectional study randomly recruited 742 premenopausal, perimenopausal, and naturally menopausal women aged 45-55 years from Beijing. Contraceptive methods were used by 75.6% of premenopausal and 54.2% of perimenopausal women. The primary methods were the IUD and barrier method. The women's choices of methods were related to parity and frequency of sexual activities. Sexual activity was related to the satisfaction of contraceptive methods. Perimenopausal and postmenopausal women were about half as likely to enjoy sexual activity and to experience orgasms than premenopausal women. Women of higher socioeconomic status had a lower risk for decreased sexual functioning. The IUD was the most popular and the most appropriate contraceptive method for perimenopausal women. Sexual function was associated with the women's satisfaction with the contraceptive method used, their menopausal status, and their socioeconomic class. PMID- 10361623 TI - A vaginal fluid simulant. AB - A fluid medium was developed to simulate the fluid produced in the human vagina. The composition of the medium was based on an extensive review of the literature on constituents of human vaginal secretions. In choosing the ingredients for this medium, the goal was to emphasize properties that influence interactions of vaginal fluid with topical contraceptive, prophylactic, or therapeutic products. Among these properties, pH and osmolarity play a dominant role in physicochemical processes that govern drug release and distribution. PMID- 10361624 TI - Use of sexually transmitted disease risk assessment algorithms for selection of intrauterine device candidates. AB - Sexually transmitted diseases (STD) are an important contraindication for intrauterine device (IUD) insertion. Nevertheless, laboratory testing for STD is not possible in many settings. The objective of this study is to evaluate the use of risk assessment algorithms to predict STD and subsequent IUD-related complications among IUD candidates. Among 615 IUD users in Kenya, the following algorithms were evaluated: 1) an STD algorithm based on US Agency for International Development (USAID) Technical Working Group guidelines: 2) a Centers for Disease Control and Prevention (CDC) algorithm for management of chlamydia; and 3) a data-derived algorithm modeled from study data. Algorithms were evaluated for prediction of chlamydial and gonococcal infection at 1 month and complications (pelvic inflammatory disease [PID], IUD removals, and IUD expulsions) over 4 months. Women with STD were more likely to develop complications than women without STD (19% vs 6%; risk ratio = 2.9; 95% CI 1.3 6.5). For STD prediction, the USAID algorithm was 75% sensitive and 48% specific, with a positive likelihood ratio (LR+) of 1.4. The CDC algorithm was 44% sensitive and 72% specific, LR+ = 1.6. The data-derived algorithm was 91% sensitive and 56% specific, with LR+ = 2.0 and LR- = 0.2. Category-specific LR for this algorithm identified women with very low (< 1%) and very high (29%) infection probabilities. The data-derived algorithm was also the best predictor of IUD-related complications. These results suggest that use of STD algorithms may improve selection of IUD users. Women at high risk for STD could be counseled to avoid IUD, whereas women at moderate risk should be monitored closely and counseled to use condoms. PMID- 10361625 TI - Side effects of mifepristone-misoprostol abortion versus surgical abortion. Data from a trial in China, Cuba, and India. AB - Although serious adverse events of early abortion have been studied, little attention has been paid to the more common side effects experienced by early medical or surgical abortion clients. Using data from a multicenter comparative trial of women < or = 56 days' gestation in China, Cuba, and India (n = 1373), side effects experienced by mifepristone-misoprostol medical abortion and surgical abortion clients were analyzed at the different stages of their abortions. Data on side effects came from women's reports at each clinic visit, providers' observations during the clinic visits, and symptom diaries maintained during the study period. Medical abortion clients at all sites experienced more side effects than their surgical counterparts. The disparity between the two groups was particularly pronounced for bleeding and pain. Despite more reports of side effects among medical abortion clients, however, assessments of well-being and reports of satisfaction at the exit interview were similar in both treatment groups. PMID- 10361627 TI - A longitudinal study of changes in endometrial microvascular density in Norplant implant users. AB - This study aimed to investigate the effects of the subdermal levonorgestrel contraceptive implant Norplant on endometrial vascular density at different durations of exposure, and the relationship between endometrial histology, vascular density, and bleeding patterns. A prospective controlled trial of Norplant implant users compared endometrial vascular density in biopsies taken before and after Norplant implant insertion. A total of 34 women with regular menstrual cycles requesting long-term contraception were recruited at the Sydney Centre for Reproductive Health Research, Australia. A significant increase in mean endometrial microvascular density was observed from as early as 3 weeks after insertion of Norplant implants. Vascular density was increased from a control secretory phase value of 189.6 (7.0 vessels/mm2 (+/- SEM) to 253.80 +/- 7 vessels/mm2 at 2-13 weeks of Norplant implant exposure (t ratio = 2.08, p = 0.01) and 212.7 +/- 12.9 vessels/mm2 at 14-42 weeks of exposure (t ratio = 2.03, p = 0.02). In those with atrophic endometrium, or in whom myometrium and basalis only were found in biopsies (20 of 66, 30%), mean endometrial vascular density was increased at 273.1 +/- 16.1 vessels/mm2 compared with 210.9 +/- 11.7 vessels/mm2 in other histological groups (F ratio = 9.74, p = 0.0028). Bleeding and spotting in the previous 30 days were less common in those with this histological appearance at a mean of 4.95 days compared with 8.22 days. This is the first study to assess endometrial vascular density in the early months of Norplant implant use. The findings suggest that the endometrial vasculature is profoundly altered in the early months of Norplant implant exposure when bleeding problems are most common. PMID- 10361626 TI - Use of a single implant of elcometrine (ST-1435), a nonorally active progestin, as a long acting contraceptive for postpartum nursing women. AB - Because of its unique features, the contraceptive effectiveness and tolerance during breast-feeding of 16-methylene-17 alpha-acetoxy-19-nor-4-pregnene-3,20 dione (elcometrine), delivered within a single subdermal capsule of medical grade polydimethylsiloxane, was investigated. Unlike other progestational steroids, elcometrine has no affinity for androgen and estrogen receptors and is inactive by the oral route. A total of 66 breast-feeding women receiving elcometrine by the subdermal route were enrolled in the study, and 69 women who elected to use Copper-T380 intrauterine devices (IUD) served as control subjects. The women and their infants were observed until the end of the first postpartum year. There were no significant differences in growth and development measurements among the infants in the elcometrine and control groups. The percentage of infants continuing to breast-feed at 3 and 6 months was significantly higher in the elcometrine group. There were no significant differences between the concentration of elcometrine in the mother's blood and milk. At 75 days, blood levels of elcometrine in the infants were near the undetectable and were significantly lower than the levels in maternal blood or milk (p < 0.01). In 15 of 25 infants, blood levels of elcometrine were at the limit of assay sensitivity or undetectable. Two pregnancies occurred in women using IUD, whereas none occurred in those using implants. There were menstrual bleeding irregularities in both groups. A single elcometrine capsule placed subcutaneously at 6-monthly intervals appears to be an effective method of contraception for lactating women and results in blood concentrations of nursing infants at or near undetectable levels. PMID- 10361628 TI - Fertilin beta peptides inhibit sperm binding to zona-free eggs in a homologous baboon in vitro fertilization system. AB - This study developed a baboon in vitro fertilization system that can be used in testing defined gamete antigens for fertility effects. A laparoscopic procedure that proved very valuable in retrieving eggs from female baboons for in vitro studies was developed. On average, 30 +/- 5 (SD) eggs were harvested per female baboon per cycle. Micromolar quantities of hexapeptides or a 28 aa residue peptide, all corresponding to fertilin beta disintegrin domain, competitively inhibited the binding of zona-free baboon eggs by baboon sperm in vitro. This study demonstrated that fertilin beta has a required role in baboon fertilization. PMID- 10361630 TI - The upper extremity trauma patient. AB - Trauma patients are predominantly young men. Urban trauma patients frequently are uninsured, and often are victims of intentionally inflicted penetrating injury, usually caused by firearms. Substance use is a common underlying factor in both urban and rural trauma. This article introduces urban and rural patients. It discusses their past medical and social histories, which are required to help the decision-making process when treating these patients. PMID- 10361631 TI - The management of penetrating trauma to the hand. AB - Penetrating gunshot wounds to the hand are becoming more common with the increase of gang violence and the availability of firearms. Hand surgeons treating victims of gunshot wounds must be familiar with the peculiarities of the penetrating missile, the special structural considerations of the hand, and the unique demands and rehabilitation needs of this difficult patient population. This article discusses penetrating trauma to the hand and includes a review of ballistics. PMID- 10361629 TI - Ultrastructural changes in the vas deferens of langur monkeys Presbytis entellus entellus after vas occlusion with styrene maleic anhydride and after its reversal. AB - Ultrastructural changes in the vas deferens of langur monkeys after 150 days of vas occlusion with styrene maleic anhydride (SMA) and after 150 days of noninvasive reversal are reported. The vas deferens of the sham-operated control animals revealed active secretory and absorptive functions. The basal cells showed prominent nucleus and sparse cytoplasmic organelles, and the principal cells characterized by oval or irregular nucleus, well developed mitochondria, Golgi bodies, rough endoplasmic reticulum, secretory granules in the Golgi area, free ribosomes, and glycogen granules in the supranuclear region suggesting secretory function. Vesicles and stereocilia in the apex region suggested absorptive functions of the vas deferens. Vas occlusion by SMA resulted in exfoliation of epithelial cells, pyknotic nuclei, and vacuolated cytoplasm virtually devoid of cytoplasmic organelles and stereocilia. After noninvasive reversal, the vas epithelium regained a state of normalcy as evidenced by prominent plasma membrane, nucleus, cytoplasmic organelles, and stereocilia. The results suggest that the exfoliation of the epithelium due to vas occlusion by SMA regains normalcy after 150 days of noninvasive reversal. PMID- 10361632 TI - Management of farm-related injuries to the upper extremity. AB - This article familiarizes physicians with common farm equipment that can cause devastating hand injuries. The focus is on farm-related injuries to the hand and stresses the degree of soft tissue injuries, discusses avulsion injuries, and acute management principles. Types of injuries, their pathophysiology, case reports, and treatments also are reviewed. This article also stresses how these injuries differ from the urban trauma patient. PMID- 10361633 TI - The treatment of forearm fractures from penetrating trauma. AB - Penetrating trauma is an endemic urban disease. The initial assessment requires a comprehensive examination and assessment of wound morphology. Preliminary splinting and radiographs of the injured extremity are necessary to plan fracture fixation. The surgical approach must appreciate the zone of injury and respect the soft tissues. Open or indirect reduction and internal fixation are the preferred techniques to stabilize the fracture. PMID- 10361634 TI - Peripheral nerve injuries secondary to missiles. AB - Peripheral nerve injuries secondary to missiles can present some of the most challenging problems faced by hand surgeons. This article reviews the pertinent neural anatomy, injury classifications, and repair techniques. Options in the management of nerve gaps are presented including the use of vascularized nerve grafts. The results are discussed and a treatment algorithm is presented. PMID- 10361635 TI - High-pressure injection injuries to the hand. AB - High-pressure injection injuries of the hand can lead to devastating outcomes. Amputation rates as high as 48% have been reported. The authors review the history of high-pressure injection injuries, with attention to the materials injected, the clinical presentation of these injuries, emergency room and subsequent definitive surgical management of these injuries, the bacteriology and recommendations for antibiotic use in these injuries, and postoperative management. PMID- 10361636 TI - Treatment of vascular injuries from penetrating and nonpenetrating trauma. AB - The treatment algorithm for vascular injury after trauma, the role of vascular studies, the role of immediate exploration, and other treatments are discussed. Case examples are provided and include direct vascular repair or a vein graft. The potential complications including compartment syndrome also is discussed. This article focuses on the trauma patient who has a vascular injury. PMID- 10361637 TI - Physical therapy after hand injuries. AB - The nuances of physical therapy necessary in the trauma patient are discussed. This article also discusses either the treatment of fractures via therapy or the treatment of nerve, tendon, or arterial injuries. It also describes physical therapy guidelines relevant to the patient with hand trauma and reviews communication between the physician and therapist in managing these patients. Intervention concepts are illustrated through case studies of patients with complex hand injuries. PMID- 10361638 TI - The role of emergency free flaps for hand trauma. AB - Primary closure of a wound with free flap requires minimizing the risk of infection in an effort to cover vital exposed structures. Careful patient selection, radical debridement of the wound, and an experienced microsurgical team are important for the routine application of this technique. Primary reconstruction of all injured structures is an extension of this technique that enables efficient management of severe upper extremity problems in a single setting. PMID- 10361639 TI - Management principles to treat nerve loss after violent trauma to the upper extremity. AB - This article discusses nerve loss treatment options. The role of cable graft versus early tendon transfers are delineated. Vascularized grafts and the timing of nerve grafting also are discussed. PMID- 10361640 TI - The management of bone defects of the forearm after trauma. AB - In summary, the authors believe that traumatic bone defects of the diaphyseal forearm, fewer than 6 cm in length, can be well managed with corticocancellous bone grafting, provided the patient has an adequate soft-tissue envelope. In cases of soft-tissue compromise, primary bone grafting is still the treatment of choice, combined with simultaneous soft-tissue coverage. This does not exclude the use of immediate primary shortening. Traumatic bone defects larger than 6 cm can best be managed with osteoseptocutaneous free fibular graft with excellent functional and cosmetic results. The creation of a one-bone forearm is rarely necessary, but remains a treatment option for an otherwise unsalvageable extremity. Osteo-articular defects can be more difficult to manage. Proximal osteo-articular defects of the radial head can be treated with excision or placement of a titanium radial head prosthesis. Distal osteo-articular defects may be better addressed in the case of radial bone loss by primary wrist fusion and, in the ulna, by a primary Darrach or Suave-Kapandji procedure. PMID- 10361641 TI - Treatment of composite tissue loss following hand and forearm trauma. AB - Restoration of satisfactory function following composite tissue loss in the hand and forearm requires reconstruction of soft tissue defect, nerve and tendon injury, and skeletal fixation when fracture occurs. Microsurgical techniques often are required, but single-donor composite tissue transfers seldom are necessary. Reconstructive strategy requires careful planning. Decision-making principles and surgical alternatives are discussed at the end of the article. PMID- 10361642 TI - The role of arthroplasty and arthrodesis following trauma to the upper extremity. AB - Intra-articular fractures in the hand and wrist are common. Prompt treatment with adequate fixation to allow early motion provides the best chance for a successful outcome. Arthrodesis and arthroplasty are the two main options for treatment of residual pain, deformity, or instability in the hand. Management of late wrist symptoms may include proximal row carpectomy, limited wrist fusion, or complete wrist fusion. PMID- 10361643 TI - Microvascular reconstruction of the traumatized thumb. AB - Microvascular reconstruction of the thumb is technically exacting and requires a firm grasp of anatomic variations of toe blood supply. Careful preoperative planning and meticulous microsurgical technique result in a low primary failure rate and maximize functional and aesthetic results. PMID- 10361644 TI - Hand infections in the trauma patient. AB - Infections are a frequent, unfortunate complication seen in orthopedic trauma patients. The specific anatomy of the upper extremity makes it an easy target for opportunistic organisms, especially after an acute traumatic event. In addition, pathogenic factors and host factors can contribute to the severity of the infection. Meticulous wound management, identification of the offending organism(s), and tailoring of care to the special needs of the very sick trauma patient limit complications. PMID- 10361645 TI - Central and peripheral adaptations after 12 weeks of exercise training in post coronary artery bypass surgery patients. AB - PURPOSE: Training adaptations in patients with coronary artery disease (CAD) have been reported previously, but little is known about central and peripheral adaptations in those recovering from coronary artery bypass graft surgery (CABG). The purpose of this study was to examine the effects of 12 weeks of endurance exercise training on exercise performance and left ventricular and peripheral vascular reserve in a group of uncomplicated CABG patients. METHODS: Thirty-one patients were recruited and began training 8 to 10 weeks after uncomplicated CABG. Patients underwent progressive exercise training consisting of walking and jogging, at 75% to 80% maximal oxygen intake (VO2max). Measures of left ventricular function included ejection fraction (EF), ventricular volumes, and the pressure volume ratio, an index of contractility. Peak ischemic exercise calf blood flow and vascular conductance was determined using strain-gauge plethysmography. Maximal oxygen intake and submaximal blood lactate concentration also was determined. RESULTS: A significant improvement in VO2max (1497 +/- 60 mL/min versus 1691 +/- 71 mL/min) was observed after training. This change was accompanied by an increase in the EF during submaximal exercise (60 +/- 3% versus 63 +/- 2% at 40% VO2max; 61 +/- 3% versus 64 +/- 3% at 70% VO2max) (P < 0.05), and the change in EF from rest to exercise (delta EF). No changes were observed for ventricular volumes during exercise, although there was a trend for a higher stroke volume at 70% VO2max. A significant increase (18%) was observed for peak ischemic exercise calf blood flow and vascular conductance. In addition, submaximal blood lactate concentration was lower after training. CONCLUSIONS: These data indicate that exercise training for 12 weeks in patients recovering from CABG can elicit significant improvements in functional capacity that, for the most part, are secondary to peripheral adaptations, with limited support for improvement in left ventricular function. PMID- 10361646 TI - The psychology of men and women recovering from coronary artery bypass surgery. AB - BACKGROUND: Little is known about the differences between men's and women's cardiac rehabilitation processes. What helps men during recovery may not necessarily aid women's recovery. Psychosocial variables are known to impact recovery in positive and negative ways. Unfortunately, it is not clear what variables are the most effective predictors of recovery outcomes for men and women. METHODS: Ninety coronary artery bypass graft patients (60 men, 30 women) completed a battery of psychological questionnaires on or after the third day after surgery. RESULTS: Results showed that women reported significantly more depressive symptoms than men. For women, pain was correlated positively with depressive symptomatology and functional impairment. For men, pain and functional impairment were correlated negatively with social support. In addition, the results of a multiple regression of pain on age, severity of disease, and two psychosocial variables (depressive symptomatology and social support) for the women showed that after controlling for age and severity of disease, depressive symptomatology and social support accounted for a significant 43% increment in the variance in pain. The psychosocial variables accounted for much less variance in pain in men. However, in a multiple regression of functional impairment on the same variables, depressive symptomatology and social support accounted for a significant 14% increment in the variance in pain in men but a nonsignificant increment for the women. CONCLUSIONS: The results support the notion that psychosocial variables play different roles in the recovery paths of men and women. In consequence, cardiac rehabilitation programs would be more effective with gender-specific tailoring. PMID- 10361647 TI - Body fat distribution's impact on physiologic outcomes during cardiac rehabilitation. AB - BACKGROUND: High waist-to-hip ratios (WHRs) predispose individuals to metabolic syndromes that may affect outcome responses to cardiac rehabilitation programs. METHODS: A total of 101 male patients who had undergone coronary artery revascularization surgery and completed 12 weeks of cardiac rehabilitation were divided into lower (LOWHR, n = 51) and higher (HIWHR, n = 50) waist-to-hip groups. Outcomes were measured at week 1 and week 12 of cardiac rehabilitation. RESULTS: Waist-to-hip ratio and body weight were greatest for HIWHR (P < 0.001) with no between-group differences in the amount of change from week 1 to 12. Triceps and subscapular skin-folds were greater for HIWHR (P < 0.001) with no difference in the amount of change between groups. Caloric expenditure during exercise class was higher for LOWHR (P = 0.022). Daily caloric expenditure was greater for LOWHR (P = 0.034) as was daily caloric intake (P < 0.001). There were no group differences for VO2peak and ventilatory anaerobic threshold (VAT) with nonsignificant trends for greater increases in LOWHR. CONCLUSIONS: Cardiac rehabilitation patients with greater WHRs expend less calories during exercise classes. To enhance overall caloric expenditure and obtain positive outcomes, cardiac rehabilitation professionals must emphasize greater activity with less sedentary time throughout the patients normal daily routine. The validity of using self-reported caloric intake and expenditure values in the cardiac rehabilitation population is questionable. PMID- 10361648 TI - Effects of a videotape information intervention at discharge on diet and exercise compliance after coronary bypass surgery. AB - BACKGROUND: This study evaluated the relative effects on compliance with recommended lifestyle changes of two experimental videotapes that involved different approaches for preparing coronary artery bypass graft (CABG) patients for the posthospital recovery period. The tapes differed in the extent to which they portrayed the recovery period as a steady, forward progression versus a series of "ups and downs." METHODS: Two hundred sixteen male and female CABG patients were assigned randomly either to view one of the two videotapes before discharge from the hospital or to receive only the standard discharge preparation provided by the hospital. All patients completed measures of anxiety and self efficacy at discharge, 1 month and 3 months after discharge from the hospital. Patients also completed measures of dietary fat consumption and activity level 1 and 3 months after discharge. RESULTS: Relative to controls, patients who viewed either of the videotapes before hospital release reported higher self-efficacy for adhering to the recommended low-fat diet both at discharge and 1 month after surgery. Viewing either of the videotapes also resulted in significantly less dietary fat intake 1 month after hospital release compared with controls. Patients who viewed the tape that portrayed the recovery period as consisting of ups and downs also reported significantly more frequent moderate exercise at 1 month and more frequent strenuous exercise 3 months after discharge. CONCLUSIONS: The experimental videotapes proved to be an effective method for increasing dietary and exercise compliance during the first 3 months after CABG. PMID- 10361649 TI - Effect of relaxation therapy on cardiac events after myocardial infarction: a 5 year follow-up study. AB - BACKGROUND: Evidence suggests that breathing and relaxation therapy may influence cardiac events in persons after acute myocardial infarction (MI). The authors studied the effects of breathing and relaxation therapy on rates of cardiac events and cost effectiveness in past MI patients. METHODS: Patients (n = 156) were chosen randomly to receive either exercise training plus relaxation therapy (relaxation group; n = 76) or exercise training only (control group; n = 80). The occurrence of major cardiac events and cardiac rehospitalizations in the two treatment groups was compared. RESULTS: At 5-year follow-up, 12 cardiac deaths had occurred, 5 in the relaxation group and 7 in the control group, reinfarction occurred in 10 and 12 patients, and cardiac surgery was performed in 2 and 11, respectively. In total, 15 (20%) and 26 (33%) patients, respectively, experienced at least one cardiac event (odds ratio [OR] for the relaxation group: 0.55, 95% confidence interval [CI] 0.29-1.05; adjusted for risk factors OR 0.52, 95% CI 0.28-0.99). Regarding all cardiac rehospitalizations, in the relaxation group, 30 patients (39%) had experienced 52 cardiac events, for which the patients were hospitalized for a total of 476 days. In the control group, 38 patients (48%) had experienced 78 cardiac events (OR 0.72; 95% CI 0.38-1.36), comprising 719 days of hospitalization. The total number of hospitalizations was reduced by 31% as a result of relaxation instruction. CONCLUSIONS: In the long-term, the disease course after myocardial infarction is influenced favorably by giving relaxation therapy in addition to cardiac rehabilitation. The extra costs of the therapy are compensated by a decrease in hospitalization for cardiac problems. PMID- 10361650 TI - Case studies of cycle exercise early after cardiothoracic surgery. PMID- 10361651 TI - Injuries and muscle soreness during the one repetition maximum assessment in a cardiac rehabilitation population. PMID- 10361652 TI - The Bone and Joint Decade, 2000-2010, for Prevention and Treatment of Musculoskeletal Disorders. PMID- 10361653 TI - Does spinal instrumentation influence the healing process of posterolateral spinal fusion? An in vivo animal model. AB - STUDY DESIGN: An in vivo sheep model was used to investigate the effect of spinal instrumentation on the healing process of posterolateral spinal fusion. OBJECTIVES: To examine the role of spinal instrumentation during the healing process of posterolateral fusion. SUMMARY OF BACKGROUND DATA: In long bone fractures, internal fixation improves the union rate but does not accelerate the healing process. Spinal instrumentation also improves the fusion rate in spinal arthrodesis. However, it remains unclear whether the use of spinal instrumentation expedites the healing process of spinal fusion. METHODS: Sixteen sheep underwent posterolateral spinal arthrodeses at L2-L3 and L4-L5 using equal amounts of autologous bone. One of those segments was selected randomly to be augmented with transpedicular screw fixation (Texas Scottish Rite Hospital spinal system). The animals were killed at 8 weeks or 16 weeks after surgery. Fusion status was evaluated by biomechanical testing, manual palpation, plain radiography, computed tomography, and histology. RESULTS: Instrumented fusion segments demonstrated significantly higher stiffness than did uninstrumented fusions at 8 weeks after surgery. Radiographic assessment and manual palpation showed that the use of spinal instrumentation improved the fusion rate at 8 weeks (47% versus 38% in radiographs, 86% versus 57% in manual palpation). Histologically, the instrumented fusions consisted of more woven bone than the uninstrumented fusions at 8 weeks after surgery. The 16-week-old fusion mass was diagnosed biomechanically, radiographically, and histologically as solid, regardless of pedicle screw augmentation. CONCLUSION: The current study's results demonstrated that spinal instrumentation creates a stable mechanical environment to enhance the early bone healing of spinal fusion. PMID- 10361654 TI - Serial changes in the rate of proteoglycan synthesis after chemonucleolysis of rabbit intervertebral discs. AB - STUDY DESIGN: Serial changes in the rate of proteoglycan synthesis in rabbit discs after chemonucleolytic treatment with chymopapain and chondroitinase ABC were measured using an in vitro method. OBJECTIVES: To determine the retained ability of the intervertebral disc to synthesize proteoglycans after chemonucleolytic treatment. SUMMARY OF BACKGROUND DATA: Most previous studies describe radiologic and histologic changes that occur after chemonucleolytic treatment. However, in humans it is not clear whether reconstitution of the disc space with normal nucleus proteoglycans can occur with time. METHODS: Twenty-five rabbits were treated with chymopapain (10 units/0.1 mL/disc) and chondroitinase ABC (5 units/0.1 mL/disc) by intradisc injection. Five rabbits were killed at each interval, 1, 2, 4, 8 and 12 weeks after injection. Radiologic changes in the disc height were noted, and the rate of proteoglycan synthesis was determined biochemically. RESULTS: After injection, no significant recovery of disc height was seen in either enzyme group after the initial disc narrowing. The average rate of proteoglycan synthesis in control rabbit intervertebral discs, those which had not been surgically treated, was 27.1 (x 10(-6) mmols sulphate/hour/dry weight). Twelve weeks after injection, the values were 21.6 in the saline group, 8.9 in the chondroitinase ABC group, and 8.2 in the chymopapain group. CONCLUSIONS: Doses within the therapeutic range can damage disc cells, at least in the rabbit, so that proteoglycan synthesis declined to 30% of control rates, and no significant recovery of disc height was observed. PMID- 10361655 TI - Action of chondroitinase ABC on epidurally transplanted nucleus pulposus in the rabbit. AB - STUDY DESIGN: After autotransplanting the nucleus pulposus into the epidural space of rabbits, chondroitinase ABC was administered, and the effect of chondroitinase ABC was examined. OBJECTIVES: To examine whether chondroitinase ABC accelerates resolution of the nucleus pulposus transplanted into the epidural space. SUMMARY OF BACKGROUND DATA: No previous reports exist on the effect of chondroitinase ABC on the nucleus pulposus in the epidural space. METHODS: In Study 1, autotransplantation of the nucleus pulposus into the epidural space was performed in rabbits. Histologic change was observed. In Study 2, dry weight, deoxyribonucleic acid content, and the amount of glycosaminoglycans of autotransplanted nucleus pulposus were quantified and compared with the respective values in the control group, chondroitinase-ABC-injected group, and phosphate-buffered saline-injected group. In Study 3, granulocytes obtained from the blood of a rabbit treated with chondroitinase ABC or phosphate-buffered saline were added to the nucleus pulposus taken from the same rabbit. RESULTS: In chondroitinase ABC group, inflammatory cells tended to infiltrate earlier than those in the control group (Study 1). The dry weight of recollected nucleus pulposus in the chondroitinase ABC group was significantly less than in the other groups. Deoxyribonucleic acid content in the nucleus pulposus tended to be larger in the chondroitinase ABC group, although no significant difference in content compared with that in the other groups was found. Regarding the residual glycosaminoglycans in the transplanted nucleus pulposus, the amount of chondroitin sulfate markedly decreased in the chondroitinase ABC group (Study 2). The number of granulocytes infiltrating the nucleus pulposus was distinctly large in the chondroitinase ABC group (Study 3). CONCLUSION: Chondroitinase ABC can enhance resolution of the nucleus pulposus in the epidural space. PMID- 10361656 TI - Characteristics of pedicle screw loading. Effect of sagittal insertion angle on intrapedicular bending moments. AB - STUDY DESIGN: A bending analysis of pedicle screws inserted into vertebral body analogues. Intravertebral and intrapedicular pedicle screw bending moments were studied as a function of sagittal insertion angle. OBJECTIVES: To determine how the pedicle screw bending moment is affected by changes in the insertion angle. SUMMARY OF BACKGROUND DATA: There is a significant incidence of failure when pedicle screws are used to instrument unstable spinal segments. Extrinsic factors that affect screw bending failure have been poorly characterized. Previous work has demonstrated that intrapedicular pedicle screw bending moments are significantly affected by the sagittal location and depth of pedicle screw placement. METHODS: Pedicle screw transducers were inserted in analogue vertebrae at one of three orientations: 7 degrees cephalad (toward the superior endplate), 7 degrees caudal (toward the inferior endplate), or parallel to the superior endplate (control). An axial load was applied to the superior endplate of the vertebra, and screw bending moments were recorded directly from the transducers. RESULTS: Screws angled 7 degrees cephalad developed significantly greater mean intrapedicular bending moments compared with screws inserted caudal or control screws. There was no significant difference in bending moments realized within the vertebral body for the three screw positions. CONCLUSIONS: Angulating pedicle screws toward the superior endplate increased bending moments within the pedicle. If attention to optimal screw insertion technique can reduce bending moments and potential for screw failure without increasing morbidity, surgical risk, or operative time, then proper insertion technique takes on new importance. PMID- 10361657 TI - Clinical validity and stability of active and passive cervical range of motion with regard to total and unilateral uniplanar motion. AB - STUDY DESIGN: A study of inter- and intra-examiner reliability and clinical validity using two instruments for assessment of spinal range of motion in healthy individuals. OBJECTIVE: To assess the clinical validity, stability, and normative values for active and passive cervical range of motion as measured by the CA-6000 (Orthopedic Systems Inc., Union City, CA), an electrogoniometer. SUMMARY OF BACKGROUND DATA: The authors' early trials with the electrogoniometer yielded values that differed substantially from those in other reports. The authors sought to resolve those discrepancies and understand their sources. METHODS: Axial rotations along the transverse, coronal, and frontal planes were measured as half-cycles (i.e., left-right or flexion-extension) that were repeated seven times per trial. Test-retest data were collected on the same healthy individuals for active and passive motion using men and women aged 20-39 years. For validity, simultaneous digital dual inclinometry and electrogoniometry were performed twice over a 1-week interval. In addition, a bench test was performed for validation of axial rotation. RESULTS: Clinical reliability of the CA-6000 was high for inter- and intra-examiner studies of total active motion, and validity was high when compared with that obtained with dual inclinometry. Total range of motion had less between-trial variability than half-cycles, axial rotation and lateral bending measurements had greater reliability than did flexion-extension measurements, and active motion was more reliable than passive motion. CONCLUSION: The CA-6000 provides valid and reliable measures of cervical range of motion. Discrepancies reported elsewhere appear to have arisen from several sources, as discussed in this article. PMID- 10361658 TI - The association between cigarette smoking and back pain in adults. AB - STUDY DESIGN: A retrospective cohort study of adolescent idiopathic scoliosis. A comparison group of persons without scoliosis was also selected randomly from the general population. OBJECTIVES: To estimate the association between level of cigarette smoking and the prevalence and severity of back pain. METHODS: A postal questionnaire was used to elicit information on smoking histories, a variety of indices of low back pain, and potential confounding factors. The association between smoking and back pain was estimated separately for men and women in the cohort and in the comparison group using ordinal regression models. RESULTS: The questionnaire was completed by 1287 women and 184 men who had adolescent idiopathic scoliosis and by 1130 women and 621 men in the comparison population who did not have scoliosis. Statistically significant associations between back pain and current cigarette smoking were found in the two groups of women and men with scoliosis, but not among men selected from the general population. In the three former groups, proportional odds ratios comparing current smokers to persons who never smoked ranged from 1.4 to 1.9. Among current smokers, the prevalence of back pain increased with cigarette consumption, and the proportional odds ratios ranged from 1.2 to 1.8 per 10 pack-years (no. of cigarettes smoked per day x no. of years/20). In these three groups, intensity, frequency, and duration of episodes of back pain also were found to increase with smoking consumption. CONCLUSION: The finding that smokers have more frequent episodes of back pain may imply that smoking exacerbates back pain, and the observation that stronger associations between back pain and smoking were found in the scoliosis cohort suggests that smoking may have a greater impact on persons with damaged spines. PMID- 10361659 TI - Cross-cultural adaptation of the Roland-Morris questionnaire for German-speaking patients with low back pain. AB - STUDY DESIGN: Cross-cultural adaptation and cross-sectional psychometric testing. OBJECTIVES: To develop and validate a cross-cultural version of the Roland-Morris Questionnaire for use in German-speaking patients with low back pain. SUMMARY OF THE BACKGROUND DATA: Clinical research related to the management of back pain would be facilitated enormously if a small number of patient-oriented questionnaires became widely used. If the transposition of a questionnaire from its original cultural context is done by simple translation, it is unlikely to be successful because of language and cultural differences. Therefore, a simple direct translation of a questionnaire from one language to another does not permit its use in clinical trials. METHODS: The instrument was translated and back-translated, pretested, and reviewed by a committee. The German version of the Roland-Morris Questionnaire was tested in 125 patients with low back pain. The study was conducted at the spa resort at Senftenberg, Austria, which is visited by patients from all countries of German-speaking Europe. Reliability and concurrent construct validity were assessed with Pearson's correlation coefficient on the Roland-Morris Questionnaire scores compared with the scales of the Medical Outcome Study Short Form-36 questionnaire. RESULTS: Pearson's correlation coefficient for test-retest reliability of the German version was r = 0.82 (P = 0.0001), and Cronbach's alpha was 0.81. The concurrent validity was r = 0.81 (Roland-Morris Questionnaire/pain rating; P = 0.0001), r = 0.48 (Roland Morris Questionnaire/forward bending; P = 0.0001), and r = -0.47 (Roland-Morris Questionnaire/lateral bending; P = 0.0001). Correlation between the functional scales of the Medical Outcome Study Short Form-36 questionnaire and the Roland Morris Questionnaire sum scores ranged from r = -0.29 (emotional limitations; P = 0.0011) to r = -0.71 (physical limitations; P = 0.0001). CONCLUSION: Because the German version of the Roland-Morris Questionnaire seems to be reliable and valid for the assessment of the functional status in German-speaking patients with low back pain, the use of this translated instrument can be recommended in future clinical trials. PMID- 10361660 TI - Evaluation of two time-specific back pain outcome measures. AB - STUDY DESIGN: Postal questionnaire to individuals with back pain. OBJECTIVE: To assess the acceptability, validity, and reliability of two existing back pain outcome measures, the Roland-Morris Questionnaire and the Von Korff scales, modified to measure the preceding 4 weeks. SUMMARY OF BACKGROUND DATA: The ideal outcome measure for studies of low back pain and disability remains elusive. Most existing measures assess current pain and disability. Measuring these factors over a preceding 4-week period may be more appropriate. METHODS: Individuals with back pain identified in a community survey were asked to complete the modified questionnaires. Validity was assessed by comparison with the Medical Outcome Study Short Form 36 and two general comparator questions on self-reported pain and disability. Repeatability was assessed using retest questionnaires. RESULTS: Completed questionnaires were returned by 95 individuals with chronic back pain. The modified Roland-Morris Questionnaire and Von Korff pain and Von Korff disability scales were completed satisfactorily by 83 (87%), 89 (94%), and 87 (92%) participants, respectively. Mean scores of the modified measures changed significantly and in a predictable manner with increasing ratings of pain and disability. They also correlated with aspects of the Medical Outcome Study Short Form 36 questionnaire. Retest data suggest that these measures are repeatable. The modified Roland-Morris Questionnaire provided adequate analyzable data only if missing values were imputed, and it explained less of the variance in the comparator questions than the modified Von Korff scales. CONCLUSIONS: The modified Von Korff scales were completed easily and appear to be valid and repeatable in this format. PMID- 10361661 TI - The effectiveness of acupuncture in the management of acute and chronic low back pain. A systematic review within the framework of the Cochrane Collaboration Back Review Group. AB - STUDY DESIGN: A systematic review of randomized controlled trials. OBJECTIVES: To evaluate the efficacy and effectiveness of acupuncture for the management of nonspecific low back pain. SUMMARY OF BACKGROUND DATA: Acupuncture is one of the oldest forms of therapy, but little is known about the effectiveness of acupuncture for low back pain. METHODS: Randomized controlled trials were done to assess the effectiveness of all types of acupuncture treatment, which involves needling for subjects with nonspecific low back pain. Two reviewers blinded with respect to authors, institution, and journal independently assessed the methodologic quality of the studies. Because data were statistically and clinically too heterogeneous, a qualitative review was performed. The evidence was classified into four levels: strong, moderate, limited, or no evidence. RESULTS: Eleven randomized controlled trials were included. Overall, the methodologic quality was low. Only two studies met the preset "high quality" level for this review. No study clearly evaluated acupuncture for acute low back pain. The results indicate that there was no evidence showing acupuncture to be more effective than no treatment. There was moderate evidence indicating that acupuncture is not more effective than trigger-point injection or transcutaneous electrical nerve stimulation, and there was limited evidence that acupuncture is not more effective than placebo or sham acupuncture for the management of chronic low back pain. CONCLUSIONS: Because this systematic review did not clearly indicate that acupuncture is effective in the management of back pain, the authors would not recommend acupuncture as a regular treatment for patients with low back pain. There clearly is a need for more high-quality randomized controlled trials. PMID- 10361662 TI - The effect of industrial back belts and breathing technique on trunk and pelvic coordination during a lifting task. AB - STUDY DESIGN: Relative phase angle was used to study segmental motion patterns during a lifting and lowering task. OBJECTIVES: To investigate the effect of back belts, breathing technique, and their interaction on lumbar and pelvic motion patterns. SUMMARY OF BACKGROUND DATA: Trunk and pelvic coordination has been investigated in healthy and low back pain populations. Back belts have been shown to alter range of motion and intra-abdominal pressure. Little has been reported about belts and coordination during lifting and lowering. Phase angle has been used for quantifying segmental coordination. METHODS: Six individuals performed lifting/lowering tasks with a 23-kg load under elastic, rigid, and no belt conditions. During a second session, individuals were trained in Valsalva's maneuver and repeated the protocol. Cinematography was used to track trunk and pelvis displacements. RESULTS: Segmental coordination during lowering generally was found to be the inverse of lifting. Significant differences in the relation between lumbar and pelvis phase angles were found during the initial stage of lifting because of the interaction of belt use and breathing. Lumbar range of motion decreased significantly with belt use during lifting and lowering. No significant change in pelvis range of motion was observed. CONCLUSIONS: Back belt use and breathing technique interacted during the initial stage of lifting to significantly alter the lumbar and pelvis phase angles. The change in segmental kinematics was similar to that previously reported for patients with a history of low back pain. Lumbar range of motion significantly decreased with belt use during both lifting and lowering. PMID- 10361663 TI - Intrathecal morphine. Double-blind evaluation of optimal dosage for analgesia after major lumbar spinal surgery. AB - STUDY DESIGN: A prospective, randomized, double-blind study. OBJECTIVES: To evaluate the efficacy and safety of three different dosages of intrathecal morphine sulfate for postoperative analgesia after lumbar spinal fusion. SUMMARY OF BACKGROUND DATA: Analgesia and respiratory depression after intrathecal morphine sulfate injection are dose related. The optimal dose to use for major spinal surgery is not known. METHODS: Sixty patients undergoing posterolateral lumbar fusion with or without decompression were divided randomly into 3 groups of 20 patients each. Anesthesia, monitoring, and surgery were similar for all patients. Just before closing of the wound, morphine sulfate was injected into the dural sack under direct visualization. Patients in groups 1-3 received 0.2 mg, 0.3 mg, and 0.4 mg morphine, respectively. Routine analgesia, consisting of diclofenac, was prescribed to use if necessary. Measurements were made and compared between the groups at zero hours (on admission to the Intensive Care Unit), 6 hours, 12 hours, 18 hours, and 24 hours after surgery. RESULTS: At zero hours and at 12 hours after surgery, pain levels were similar in groups 2 and 3, but were significantly higher in group 1 (P < 0.05). The respiratory rate was significantly lower in group 3 than in the other two groups (P < 0.05), and the arterial CO2 tension (PaCO2) was consistently higher in group 3. PaCO2 decreased in all three groups over the first 24 hours. Pruritus and nausea did not differ among the three groups. CONCLUSIONS: For adult patients undergoing posterolateral lumbar fusion, 0.3 mg (0.004 mg/kg) is probably the optimal dose of intrathecal morphine to manage pain. PMID- 10361664 TI - Hardware failure in an unconstrained lumbar pedicle screw system. A 2-year follow up study. AB - STUDY DESIGN: A consecutive study of patients who underwent lumbar spinal arthrodesis with an unconstrained pedicle screw system. OBJECTIVES: To determine the rate of arthrodesis and of clinical success and to examine and characterize the cases of hardware failure with the AO/Dynamic Compression Plate system (Synthes, Paoli, PA). SUMMARY OF BACKGROUND DATA: Although the advantages and disadvantages of nonconstrained versus constrained systems have been studied extensively, instrumentation failure has not. Additionally, the association between pseudarthrosis and hardware failure per se is unclear. METHODS: Seventy four consecutive cases of lumbar spinal fusion are reviewed. Standard outcome scores based on pain relief and medication usage were tabulated, along with pertinent demographic data. The patients were observed at five intervals after surgery for at least 2 years (range, 24 to 35 months; mean, 27 months). Standard statistical analyses were used to analyze data. Status of the arthrodesis was determined by standard radiographic criteria. RESULTS: The overall fusion rate was 61%. At final follow-up, 60% of patients believed that their back pain had improved, whereas 70% believed that their limb pain had improved. The presence of a solid fusion (r = 3.3, P = 0.010) was correlated positively with a successful clinical outcome; the presence of pseudarthrosis and preoperative narcotic use were negatively correlated with a successful clinical outcome. Twenty-two percent of patients (16) experienced hardware failure. Twelve of the 16 had pseudarthrosis; in the majority of these patients, hardware failure occurred at the level of the pseudarthrosis. CONCLUSIONS: The results of this study demonstrate an extremely high rate of hardware failure and pseudarthrosis using an unconstrained pedicle screw system. Interestingly, the initial rate of pain relief was higher and declined over time and was quite possibly associated with loosening of the hardware. Based on these data, it is difficult to recommend the use of an unconstrained fixation system in the lumbar spine. PMID- 10361665 TI - Neurologic compromise after an isolated laminar fracture of the cervical spine. AB - STUDY DESIGN: Report of a rare fracture of the cervical spine. OBJECTIVES: To illustrate the importance of the cervical spinolaminar line in the diagnosis of this unusual injury and to comment on appropriate investigations, management, and outcome. SUMMARY OF BACKGROUND DATA: Laminar fractures of the cervical spine are uncommon and are often missed. They usually occur after a hyperextension injury. It is unusual for these injuries to cause neurologic compromise. The injury reported here differs in that it was a result of direct trauma to the posterior aspect of the neck, and there was a significant neurologic deficit. METHODS: The clinical findings, roentgenographic appearance, treatment, complications, and follow-up assessment are presented and discussed. RESULTS: Initial neurologic examination revealed a right hemiparesis. Radiographs showed disruption of the spinolaminar line at C5 and a computed tomography scan revealed a fracture of the lamina of C5 with spinal canal encroachment. Management included high-dose corticosteroid administration and a posterior spinal decompression. The patient's initial postoperative course was complicated by acute pulmonary edema, which responded well to intravenous Furosemide and ventilation. Follow-up assessment showed significant neurologic improvement. CONCLUSIONS: The satisfactory outcome in the case of this rare injury was the result of a prompt, accurate diagnosis and appropriate management. PMID- 10361666 TI - Membranocystic lesion in lumbar yellow ligament. AB - STUDY DESIGN: An examination of surgical cases of membranocystic lesions in the lumbar yellow ligament between the fourth and fifth lumbar vertebrae. OBJECTIVES: To report the incidence and pathogenesis of membranocystic lesions of the yellow ligament in surgical specimens. SUMMARY OF BACKGROUND DATA: The membranocystic lesion has been observed not only in membranous lipodystrophy, but also in other conditions. However, there has been no report concerning this lesion in the yellow ligament. METHODS: Forty-four yellow ligaments excised in surgery were histologically reviewed. In eight cases, S-100 expression was investigated. In two cases, ultrastructural findings were examined. RESULTS: Membranocystic lesions were present in 8 (18%) of the 44 cases. The cases with the lesions had undergone surgery for spondylolisthesis (4 cases), sequestration-type disc herniation (2 cases), postradiation status (1 case), and spinal stenosis (1 case). Histologically, in all 8 cases, fibrosis was present around the lesion. In 5 of the 8 cases, chondrocytes were observed adjacent to the lesion, and nuclei positive for S-100 protein were observed in 4 cases. Nuclei of chondrocytes adjacent to the lesion also were positive for S-100 protein. Ultrastructurally, irregularly shaped cystic spaces surrounded by an electron-dense membranous structure, which were identical to those of membranous lipodystrophy, were observed. CONCLUSIONS: These results suggest that degeneration of the chondrocytes has a causal relationship to the formation of membranocystic lesions in the yellow ligament. PMID- 10361667 TI - Seat belt fracture with late development of an enterocolic fistula in a child. A case report. AB - STUDY DESIGN: A case report of a 9-year-old boy treated at a pediatric trauma center for a flexion-extension spiral fracture with late development of an enterocolic fistula subsequent to a high-velocity motor vehicle accident. OBJECTIVES: To increase the awareness of possible delayed bowel complications associated with flexion-distraction injuries of the spine in children. SUMMARY OF BACKGROUND DATA: Flexion-distraction fractures of the spine in children wearing lap seat belts, so-called "Chance" fractures, are an increasingly common result of high-velocity collisions. This type of fracture, referred to as a seat-belt fracture, is often associated with duodenal or jejunal tears. Although such intra abdominal injuries are common in such fractures secondary to this type of trauma, the occurrence of an enterocolic fistula has never been reported. METHODS: A review of all pediatric Chance fractures managed at the Children's Hospital of Eastern Ontario, as well as a literature review of all reported series of flexion distraction injuries to the spine in children, were performed. RESULTS: The subtle and prolonged symptomatology of this lesion and its similarity to a cast syndrome is emphasized. CONCLUSION: Because the orthopedic surgeon is usually the primary care-giver for children with this type of seat-belt trauma, an appreciation of the possibility of a delayed onset enterocolic fistula with its symptomatology is essential to avoid prolonged morbidity. PMID- 10361668 TI - Metastatic cardiac angiosarcoma of the cervical spine. Case report. AB - STUDY DESIGN: A case report of metastatic cardiac angiosarcoma of the cervical spine. OBJECTIVES: To show that this rare spine tumor behaves in the same manner as an arteriovenous malformation and embolization, which can allow for successful spine surgery, and to discuss the natural history and rarity of this tumor. SUMMARY OF BACKGROUND DATA: Primary angiosarcoma of the heart is a very rare tumor, with fewer than 200 reports in the English literature and nothing reported in the spine literature. RESULTS: The patient in this study initially sought treatment for neck pain, left arm pain, and weakness 17 months after cardiac surgery and subsequent chemotherapy. A cervical computed tomography scan demonstrated a C5 lytic vertebral body tumor with intracannilicular extension and cord compression. An anterior cervical approach was made, but the tumor was too vascular to resect, and surgery was aborted. The C5 vascular vertebral body metastasis subsequently was embolized successfully by an interventional neuroradiologist. Reoperation via an anterior approach with corpectomy, cadaveric fibula, and anterior locking plate internal fixation was successful, producing marked improvement in the patients' symptoms. CONCLUSION: Spinal involvement by primary cardiac angiosarcoma is very rare, and this is only the second operative case ever reported. The vascular nature of this tumor makes it behave in a manner similar to that of a high-flow arteriovenous malformation. Surgery should not be undertaken before preoperative angiography and embolization. The dismal prognosis for this rare malignancy is discussed. PMID- 10361669 TI - The modern hippocratic tradition. Some messages for contemporary medicine. AB - Hippocrates (5th century B.C.), the most prominent physician of antiquity, was born in the small Greek island of Kos, which is near the coast of Asia Minor. Before his era, medicine was practiced as an empirical art and had a religious nature. Hippocratic medicine represents the landmark for the evolution of Western medicine. This "father" of rational medicine assimilated the accumulated knowledge of the past and formed a diagnostic system based on clinical observation and logical reasoning. The great physician attributed diseases to natural causes, believed in the healing power of nature, and gave special emphasis to the prevention and prognosis of illnesses. He treated patients as psychosomatic entities (a holistic medical approach) in relation to their natural environment. In his treatises, Hippocrates defined the ethical principles guiding medical practice. His entire work was inspired by humanistic ideals and an undeviating dedication to the patient. Modern medicine can derive valuable lessons from the Hippocratic tradition. For the coming 21st century, medicine more than ever senses the need to combine the concepts of humanistic values and the Hippocratic messages with the technologic "imperative" (power). This bond is necessary to the improvement of medicine in the future because, currently, the enormous biomedical technology so far has contributed little to the traditionally human fields of psychosomatic and functional disturbances, posing new dilemmas and threatening scientific problems. PMID- 10361670 TI - The influence of occupation on lumbar degeneration. AB - In many countries, back problems have been defined as occupational injuries. The belief underlying this injury model is that back symptoms are caused primarily by work-related mechanical factors that damage the structures of the spine, either through a single incident or repeated loading. Although the etiopathogenesis of degenerative findings in the disc and their relation to pain are poorly understood, changes in the disc are suspected of underlying many back symptoms. The focus of this article is on examining the relation between occupational factors and disc degeneration. Occupational factors suspected of accelerating spinal degeneration include accident-related trauma; heavy physical loading and materials handling, including lifting, bending, and twisting; prolonged sitting; and sustained nonneutral work postures and vehicular driving. There is evidence to suggest that occupational exposures have an effect on disc degeneration. However, these factors explain little of the variability in degeneration found in the adult population. Furthermore, the lack of a clear dose-response relation between time spent in various occupational loading conditions and degenerative findings adds to doubts about a strong causal link. The contribution of suspected occupational risk factors appears to be particularly modest when compared with familial influences, which reflect the combined effects of genes and early childhood environment. These findings challenge the dominant role assumed for occupational loading in disc degeneration and associated back problems, and suggest a more complex etiology. PMID- 10361671 TI - Somatosensory- and motor-evoked potential monitoring without a wake-up test during idiopathic scoliosis surgery: an accepted standard of care. PMID- 10361672 TI - Cervical epidural steroid injection with intrinsic spinal cord damage. PMID- 10361673 TI - Whiplash associated disorders: redefining whiplash and its management. PMID- 10361674 TI - Time-dependent changes of amino acids in the serum, liver, brain and urine of rats administered with theanine. AB - Time-dependent changes of theanine (gamma-glutamylethylamide) and other amino acids in various tissues of rats were investigated during the 24 hrs after theanine administration. When theanine (4 g/kg of body weight) was intragastrically administered to rats, the concentrations of theanine in the serum, liver and brain were significantly increased 1 hr after its administration, and thereafter gradually decreased, but reached the maximum level in the brain after 5 hrs. Theanine in these tissues had completely disappeared 24 hrs after its administration. In contrast, the administration of theanine resulted in the concentrations of theanine, urea, ethylamine and glutamic acid in the urine being significantly enhanced. These results suggest that theanine might be degraded via glutamic acid. PMID- 10361675 TI - Complementary effects of bifidogenic growth stimulators and ammonium sulfate in natural rubber serum powder on Bifidobacterium bifidum. AB - Natural rubber serum powder, rich in crude protein and carbohydrates, had a strong growth-stimulating activity for Bifidobacterium bifidum JCM 1254, which was unable to grow in a fully synthetic medium, B12 assay medium. Natural rubber serum powder was fractionated by ultrafiltration (molecular weight cutoff 1000). The active ultrafiltrate was further concentrated and desalted with an adsorptive microconcentrator, which adsorbs virtually all amino acids and peptides. Through this purification step, it was found that the adsorbed fraction obtained did not stimulate growth independently but acted complementarily with a small amount of ammonium sulfate. The adsorbed fraction was subsequently analyzed on reversed phase high pressure liquid chromatography, and the activities of the eluates were measured on B12 assay medium with ammonium sulfate. Consequently, it was proved that several peptidic ingredients in the adsorbed fraction increased the growth of B. bifidum. PMID- 10361676 TI - Two cytosolic cyclophilin genes of Arabidopsis thaliana differently regulated in temporal- and organ-specific expression. AB - We have previously isolated two closely related genes (ATCYP1 and ATCYP2) each encoding a cytosolic cyclophilin of Arabidopsis thaliana. Here we tested expression patterns of these two genes by Northern analysis and by histochemical analysis with transgenic plants carrying the promoter: beta-glucuronidase (GUS) fusion. The results showed that ATCYP1 is predominantly transcribed in vascular tissue and flowers, but ATCYP2 is at higher levels in younger leaves. The different expression patterns seemed to be conferred by the quite different promoter structures carrying various cis elements. Our finding suggests that the two cyclophilins have different roles in Arabidopsis thaliana cells. PMID- 10361677 TI - Preparation and application of anti-idiotypic antibody against anti-gibberellin A4 antibody. AB - A monoclonal anti-idiotypic antibody was raised against anti-gibberellin A4 (GA4) antibody, which recognizes biologically active gibberellins such as GA1 and GA4 specifically. Amino acid sequences of variable regions of both anti-GA4 and anti idiotypic antibodies were analyzed. By using the property of the anti-idiotypic antibody to compete with GA1/4 in binding to the anti-GA4 antibody, we successfully applied the anti-idiotypic antibody to ELISA as a tracer for measuring GA1/4. The single-chain Fv (scFv) gene of the anti-idiotypic antibody was constructed, and scFv expressed in E. coli showed binding activity to anti GA4 antibody. These results suggest the possible application of anti-idiotypic antibody as a handy and stable source of an enzymatic tracer for ELISA by production of fusion protein of the scFv and an appropriate enzyme. PMID- 10361678 TI - Effects of DFA IV in rats: calcium absorption and metabolism of DFA IV by intestinal microorganisms. AB - Di-D-fructose-2,6':6,2'-dianhydride (DFA IV) is a disaccharide consisting of two fructose residues that can be prepared from levan by levan fructotransferase from Arthrobacter nicotinovorans GS-9, and it can be expected to have novel physiological functions from its unique structure. In this study, the effects of DFA IV on calcium absorption and the metabolism of DFA IV by intestinal microorganisms were studied in rats to examine the physiological functions of DFA IV. The apparent calcium absorption in rats fed with DFA IV was significantly higher than that in the control rats, and it seems that calcium absorption had almost been completed at the end of the small intestine. DFA IV also increased the calcium absorption in in vitro experiments, using everted jejunal and ileal sacs, and this result supports the finding obtained in the in vivo experiments. These results indicate that DFA IV may have a function for increasing the calcium absorption in the small intestine of rats. However, the effect in the large intestine could not be clearly observed because of the lack of calcium that reached there. The results of analyses of organic acids in the cecal and colonic contents and of DFA IV in the fecal, cecal, and colonic contents showed that the metabolism of DFA IV by microorganisms in the large intestine progressed gradually, and that DFA IV was converted mainly to acetate, butyrate, and lactate. PMID- 10361679 TI - Expression of elongation factor 1 beta' in Escherichia coli and its interaction with elongation factor 1 alpha from silk gland. AB - Silk gland elongation factor 1 (EF-1) consists of four subunits: alpha, beta, beta', and gamma. EF-1 beta beta' gamma catalyzes the exchange of GDP for GTP on EF-1 alpha and stimulates the binding of EF-1 alpha-dependent aminoacyl-tRNA to ribosomes. The carboxy-terminal regions of the EF-1 beta subunits from various species are highly conserved. We examined the region of EF-1 beta' that binds to EF-1 alpha by in vitro binding assays, and examined the GDP/GTP exchange activity using deletion mutants of a GST-EF1 beta' fusion protein. We thereby suggested a pivotal amino acid region, residues 189-222, of EF-1 beta' for binding to EF-1 alpha. PMID- 10361680 TI - Construction of an L-isoleucine overproducing strain of Escherichia coli K-12. AB - The genes for a threonine deaminase that is resistant to feedback inhibition by L isoleucine and for an active acetohydroxyacid synthase II were introduced by a plasmid into a L-threonine-producing recombinant strain of Escherichia coli K-12. Analysis of culture broth of the strain using 13C nuclear magnetic resonance suggested that alpha, beta-dihydroxy-beta-methylvalerate (DHMV) and alpha-keto beta-methylvalerate (KMV), the third and the fourth intermediates in the L isoleucine biosynthetic pathway from L-threonine, respectively, accumulated in the medium in amounts comparable to that of L-isoleucine. The ratio of accumulated L-isoleucine:DHMV:KMV were approximately 2:1:1. The concentration of accumulated L-isoleucine increased by twofold after the additional introduction of the genes for dihyroxyacid dehydratase (DH) and transaminase-B (TA-B), and the intermediates no longer accumulated. The resultant strain TVD5 accumulated 10 g/l of L-isoleucine from 40 g/l of glucose. PMID- 10361682 TI - Elevated body fat in rats by the dietary nitric oxide synthase inhibitor, L-N omega nitroarginine. AB - The influence of the dietary nitric oxide (NO) synthase inhibitor, L-N omega nitroarginine (L-NNA) on body fat was examined in rats. In experiment 1, all rats were fed with the same amount of diet with or without 0.02% L-NNA for 8 wk. L-NNA intake caused elevations in serum triglyceride and body fat, and reduction in serum nitrate (a metabolite of nitric oxide). The activity of hepatic carnitine palmitoyltransferase was reduced by L-NNA. In experiment 2, rats were fed for 8 wk with the same amount of diets with or without 0.02% L-NNA supplemented or not with 4% L-arginine. The elevation in body fat, and the reductions in serum nitrate and in the activity of hepatic carnitine palmitoyltransferase by L-NNA were all suppressed by supplemental L-arginine. The results suggest that lower NO generation elevated not only serum triglyceride, but also body fat by reduced fatty acid oxidation. PMID- 10361681 TI - A new carboxylesterase from Brevibacterium linens IFO 12171 responsible for the conversion of 1,4-butanediol diacrylate to 4-hydroxybutyl acrylate: purification, characterization, gene cloning, and gene expression in Escherichia coli. AB - A carboxylesterase that is responsible for conversion of 1,4-butanediol diacrylate (BDA) to 4-hydroxybutyl acrylate (4HBA) was found in Brevibacterium lines IFO 12171, and purified to homogeneity. The purified enzyme was active toward a variety of diesters of ethylene glycol, 1,4-butanediol, and 1,6 hexanediol. The K(m) and kcat of the enzyme for BDA were 3.04 mM and 203,000 s-1, respectively. The reaction with the purified enzyme gave 98 mM 4HBA from 100 mM BDA for 60 min. The enzyme gene was cloned from the chromosomal DNA of the bacterium. The open reading frame encoding the enzyme was 1176 bp long, corresponding to a protein of 393 amino acid residues (molecular mass = 42,569 Da). The deduced amino acid sequence contained the tetra peptide motif sequence, STTK, and the serine residue was confirmed to be the catalytic center of BDA esterase by site-directed mutagenesis for several amino acid residues. The gene was expressed in Escherichia coli under the control of the lac promoter, and the gene product (a fusion protein with 6 amino acid residues from beta galactosidase) showed the same catalytic properties as the enzyme from the parent strain. PMID- 10361683 TI - Production of recombinant Der fI (a major mite allergen) by Aspergillus oryzae. AB - Der fI is a major mite allergen. To produce Der fI by Aspergillus oryzae, we placed a DNA fragment encoding precursor-type recombinant Der fI E(-1)K (reDer fI E(-1) K), which had the C-terminal amino acid of the pro-sequence (Glu) changed to Lys, downstream of the glaA gene promoter and introduced it into Aspergillus oryzae. In liquid culture, most of the reDer fI E(-1)K produced by the transformants was degraded when culture was shaken vigorously. However, the degradation of reDer fI E(-1)K was suppressed when it was shaken gently. The processed reDer fI E(-1)K could be obtained after lysylendopeptidase and endoglycosidase Hf (Endo Hf) treatment. The yield of processed reDer fI E(-1)K was 8 mg/l. When the transformant was grown on a wheat bran culture, the yield of processed reDer fI E(-1)K reached 48 mg/kg. Because processed reDer fI E(-1)Ks obtained from both cultures had almost the same IgE-binding activity and elicited the same skin reaction as native Der fI, they could be very useful for diagnostic purposes or immunotherapy. PMID- 10361684 TI - High-multiplicity of chitinase genes in Streptomyces coelicolor A3(2). AB - Six different genes for chitinase from ordered cosmids of the chromosome of Streptomyces coelicolor A3(2) were identified by hybridization, using the chitinase genes from other Streptomyces spp. as probes, and cloned. The genes were sequenced and analyzed. The genes, together with an additional chitinase gene obtained from the data bank, can be classified into either family 18 or family 19 of the glycosyl hydrolase classification. The five chitinases that fall into family 18 show diversity in their multiple domain structures as well as in the amino acid sequences of their catalytic domains. The remaining two chitinases are members of family 19 chitinases, since their C-terminus shares more than 70% identity with the catalytic domain of ChiC of Streptomyces griseus, the sole gene for family 19 chitinase so far found in an organism other than higher plants. PMID- 10361685 TI - Phloretin-induced apoptosis in B16 melanoma 4A5 cells and HL60 human leukemia cells. AB - The dihydrochalcone phloretin induced apoptosis in B16 mouse melanoma 4A5 cells and HL60 human leukemia cells. Phloretin was suggested to induce apoptosis in B16 cells mainly through the inhibition of glucose transmembrane transport. The phloretin-induced apoptosis in B16 cells was inhibited by actinomycin D, Ac-YVAD CHO caspase-1-like inhibitor, and Ac-DEVD-CHO caspase-3-like inhibitor. During the induction of apoptosis by phloretin, the expression of Bax protein in B16 cells increased and the levels of p53, Bcl-2, and Bcl-XL proteins did not change. Our results suggested that phloretin induced apoptosis through the promotion of Bax protein expression and caspases activation. On the other hand, phloretin may induce apoptosis in HL60 cells through the inhibition of protein kinase C activity because phloretin inhibited protein kinase C activity in HL60 cells more than that in B16 cells. The phloretin induced-apoptosis in HL60 cells was not inhibited by actinomycin D and the caspase-1-like inhibitor, but slightly inhibited by the caspase-3-like inhibitor. Phloretin reduced the level of caspase 3 protein in HL60 cells, but not the level of the Bcl-2 protein. Phloretin did not increase the level of Bax protein. Phloretin was suggested to induce apoptosis in HL60 cells through the inhibition of protein kinase C activity, followed by the pathway, which is different from that in B16 cells. PMID- 10361686 TI - (2E,6R)-8-hydroxy-2,6-dimethyl-2-octenoic acid, a novel anti-osteoporotic monoterpene, isolated from Cistanche salsa. AB - (2E,6R)-8-Hydroxy-2,6-dimethyl-2-octenoic acid [(R)-HDOA], a novel monoterpene from Cistanche salsa, a Chinese herb, was found to be an anti-osteoporotic compound. The extract of Cistanche salsa significantly suppressed the bone weight loss in ovariectomized mice, a postmenopausal osteoporosis model. The active substance was then purified by using this osteoporotic model and the chemical structure was determined. The active compound from Cistanche salsa, (R)-HDOA, suppressed the decrease of bone weight and the mechanical strength in the ovariectomized mice. Furthermore, (R)- and (S)-HDOA were synthesized and the activity of each was evaluated. (R)-HDOA suppressed the bone weight loss, although (S)-HDOA did not showed any activity. PMID- 10361687 TI - Potentiation of GABAA receptors expressed in Xenopus oocytes by perfume and phytoncid. AB - To study the effects of perfume and phytoncid on GABAA receptors, ionotropic GABAA receptors were expressed in Xenopus oocytes by injecting mRNAs that had been prepared from rat whole brain. Essential oil, perfume and such phytoncid as leaf alcohol, hinokitiol, pinene, eugenol, citronellol and citronellal potentiated the response in the presence of GABA at low concentrations (10 and 30 microM), possibly because they bound to the potentiation-site in GABAA receptors and increased the affinity of GABA to the receptors. Since it is known that the potentiation of GABAA receptors by benzodiazepine, barbiturate, steroids and anesthetics induces the anxiolytic, anticonvulsant and sedative activity or anesthetic effect, these results suggest the possibility that the intake of perfume or phytoncid through the lungs, the skin or the intestines modulates the neural transmission in the brain through ionotropic GABAA receptors and changes the frame of the human mind, as alcohol or tobacco does. PMID- 10361688 TI - Effect of media constituents on the formation by halophilic yeast of the 2 (or 5) ethyl-5 (or 2)-methyl-4-hydroxy-3 (2H)-furanone aroma component specific to miso. AB - The formation of HEMF [2 (or 5)-ethyl-5 (or 2)-methyl-4-hydroxy-3 (2H)-furanone] by yeast was examined in an attempt to investigate its mechanism and involved factors. HEMF formation was promoted by yeast cultivation in a heat-sterilized medium which included glucose, ribose, and a nitrogenous compound such as an extract of shoyu koji, poly-peptone, casamino acid, or an amino acid (glutamic acid, threonine, serine, or alanine). PMID- 10361689 TI - Improved suppression by dietary taurine of the fecal excretion of bile acids from hypothyroid rats. AB - The effect of dietary taurine, 2-aminoethanesulfonic acid, on hypercholesterolemia caused by thiouracil-induced hypothyroidism was investigated in hypothyroid rats. Serum total- and HDL-cholesterol were significantly increased, and the excretion of fecal bile acids was significantly decreased. Taurine did not change the hypercholesterolemia, but significantly recovered the excretion of bile acids. PMID- 10361690 TI - Isolation and structural determination of a novel bicyclic taxane diterpene from needles of the Chinese yew, Taxus mairei. AB - A novel bicyclic taxane diterpene with a rare 12-membered ring was isolated from needles of the Chinese yew, Taxus mairei, and its structure was established as (3E, 8E)-2 alpha, 7 beta, 9, 10 beta, 13 alpha, 20-hexaacetoxy-5(2'-acetoxy cinnamoyloxy)-3,8-secotaxa -3,8,11-triene (1) with the help of 1D and 2D NMR data. The relative stereochemistry was deduced from a NOESY experiment. PMID- 10361691 TI - Differential accumulation of transcripts encoding sulfur assimilation enzymes upon sulfur and/or nitrogen deprivation in Arabidopsis thaliana. AB - Expression of nine genes encoding enzymes involved in the sulfur assimilation pathway was examined by RNA blot hybridization. Significantly increased levels of transcripts encoding ATP sulfurylase and APS reductase were apparent under sulfur deprivation. However, in the absence of nitrogen, their responsiveness to sulfur deprivation was markedly reduced. Results suggest that the sulfur assimilation pathway is regulated at the transcriptional level by both nitrogen and sulfur sources. PMID- 10361693 TI - Detection and analysis of translation elongation factor 3 genes from various yeasts. AB - Yeast translation requires a unique elongation factor, EF-3. However, information about EF-3 genes has been limited to only a few yeast species. Here, we developed a PCR-based system to detect the EF-3 genes specifically, and identified EF-3 gene fragments from various yeast species in which EF-3 genes have not yet been found. PMID- 10361692 TI - Sequence of a cDNA encoding mouse F1F0-ATP synthase g subunit. AB - A pregnant-induced clone was identified by differential screening from a cDNA library of mouse mammary gland. The clone was identified as a full-length cDNA encoding the F1F0-ATP synthase g subunit. Comparison of the deduced amino acid sequences of mouse ATP synthase g subunit with those of bovine species showed 86% identity. The high levels of ATP synthase g subunit mRNA were detected in heart and uterine tissues. PMID- 10361694 TI - Construction and characterization of Escherichia coli disruptants defective in the yaeM gene. AB - Escherichia coli disruptants defective in the yaeM gene, which is located at 4.2 min on the chromosome map, were constructed and characterized. The disruptants showed auxotrophy for 2-C-methylerythritol, a free alcohol of 2-C-methyl-D erythritol 4-phosphate that is a biosynthetic precursor in the nonmevalonate pathway. This result clearly shows that the yaeM gene is indeed involved in this pathway in E. coli. PMID- 10361695 TI - Expression of a functional single-chain antibody against GA24/19 in transgenic tobacco. AB - An anti-gibberellin A24/19 single-chain Fv gene was constructed from gamma and kappa genes cloned from a hybridoma cell line producing monoclonal antibody against gibberellin A24/19, biosynthetic precursors of gibberellin A4/1 which are biologically active per se. The single-chain Fv gene was introduced into tobacco plants after the binding activity of the single-chain Fv expressed in Escherichia coli was confirmed. When the single-chain Fv expression is targeted to endoplasmic reticulum, the plants could accumulate the single-chain Fv protein with the antigen binding activity up to 3.6% of the total soluble protein. On the other hand, when the expression is targeted to cytosol, accumulation of the single-chain Fv protein was not detected at all. The dwarf phenotype of the transgenic plants expressing the single-chain Fv protein, together with the preliminary analytical data indicating a decreased level of gibberellin A1 in the dwarf transgenics, suggested that the single-chain Fv decreased the concentration of bioactive gibberellins by trapping and inhibiting the metabolism of gibberellin A24 and/or A19 to gibberellin A4 and/or A1. PMID- 10361696 TI - Synthesis and cytotoxic activity of 1-alkoxy- and 1-amino-2-hydroxy-1,2 dihydroacronycine derivatives. AB - Sixteen new derivatives of the natural alkaloid acronycine, bearing 1-alkoxy or 1 amino and 2-hydroxy groups, were synthesized in order to clarify the role of the C-1 substitution. Studies on the cytotoxic activity of compounds 4-19 were carried out in vitro on L-1210 cells. Structure-activity relationships are discussed. PMID- 10361697 TI - Regiospecific synthesis of pyrido[3,4-b]- and pyrido[4,3-b]carbazole-5,11-dione derivatives. Evaluation of their in vitro antifungal or antiprotozoological activities. AB - Hetero Diels-Alder reactions between 2- or 3-bromocarbazolequinones 1a or 1b and azadiene 5 afford regiospecifically pyrido[3,4-b]- and pyrido[4,3-b]carbazole 3,5,11-triones 6a and 6b. The regiochemistry of the cycloadditions is controlled by the position of the bromine atom at C-2 or C-3 of the bromoquinone. The corresponding N- and O-methyl derivatives 7 and 8 are prepared. Structural assignment of the regioisomers is made by 1H-NMR nuclear Overhauser effect difference experiments performed on a diacetoxy derivative of pyrido[4,3 b]carbazole 9b. The in vitro antifungal and antiprotozoological activities of some prepared derivatives have been evaluated against Candida albicans, Candida krusei, Cryptococcus neoformans and Trichomonas vaginalis. None of the tested compounds have shown significant activity towards the yeasts or protozoa. PMID- 10361699 TI - Identification of photolabeled peptides for the acceptor substrate binding domain of beta 1,4-galactosyltransferase. AB - We successfully applied a carbene-generating N-acetylglucosamine derivative carrying a biotinyl group to the radioisotope-free identification of peptides within bovine UDP-galactose: N-acetylglucosamine beta 1,4-galactosyltransferase (GalT, EC 2.4.1.38) catalytic domain. Owing to the low yield of cross-linking, conventional photoaffinity labeling experiments usually encounter a thorny problem in attempting to isolate labeled components from very complex mixtures. A biotin tag introduced with our photoaffinity probe enabled us to separate the photolabeled protein from a large amount of coexisting unlabeled GalT. The introduction of biotin was also useful for the radioisotope-free detection of a labeled protein based on a highly sensitive chemiluminescent technique. We developed a novel poly(vinylidene difluoride) membrane for the identification of labeled peptides in a simple dot blot assay. Using this membrane, we successfully identified biotinyl peptides among a number of HPLC separated fragments derived from the protease digestion of photolabeled GalT proteins. The sequence analysis revealed that the biotin tag was incorporated within a tryptic GalT fragment of Y197-R208. Our approach yields, for the first time, information on the acceptor substrate binding-site fragment in this enzyme, that has been difficult to obtain using other approaches. These data are consistent with previous suggestions concerning the GalT acceptor site and clearly demonstrate the effectiveness of our approach for rapid identification of photolabeled peptides. PMID- 10361700 TI - Synthesis and biological evaluation of 1,2,3,4-tetrahydroisoquinoline derivatives as potent and selective M2 muscarinic receptor antagonists. AB - A series of 1,2,3,4-tetrahydroisoquinoline derivatives containing the 5,11 dihydro-6H-pyrido[2,3-b][1,4]benzodiazepin-6-one skeleton were prepared and evaluated for their in vitro binding affinities to muscarinic receptors and for antagonism of bradycardia in vivo. Among them, compound 3f had the highest affinity for M2 muscarinic receptors in the heart (pKi = 9.1) with low affinity for M3 muscarinic receptors in the submandibular gland. A structure-activity relationship (SAR) study suggested that the benzene ring fused piperidine and the alkyl linker chain length are crucially important for increased M2 affinity. PMID- 10361701 TI - HPLC profile analysis of hepatoprotective oleanene-glucuronides in Puerariae Flos. AB - In order to confirm the constitution of hepatoprotective oleanene glucuronide (OG), HPLC profile analyses of the total OG fractions of both Puerariae Thomsonii Flos (the flowers of Pueraria thomsonii) and Puerariae Lobatae Flos (the flowers of P. lobata) were performed. No remarkable difference in the HPLC profiles with respect to OGs in the flowers was shown, in contrast to those of the roots. By repeated chromatography of the total OG fraction of Puerariae Thomsonii Flos, soyasaponin I (1), kaikasaponin III (2) and kakkasaponin I (3), which had been already isolated from Puerariae Lobatae Flos, were obtained. The hepatoprotective activity of 2 towards immunologically induced liver injury was significantly more effective than that of 1. This information supported previously obtained structure-hepatoprotective relationship data which was measured on another model. The structure-activity relationship information which suggested that the hydroxymethyl group of the galactosyl unit would enhance the hepatoprotective activity was also substantiated. PMID- 10361702 TI - Biotransformation of hinesol isolated from the crude drug Atractylodes lancea by Aspergillus niger and Aspergillus cellulosae. AB - Biotransformation of the sesquiterpene alcohol hinesol (1) with spasmolytic activity, which was prepared from the rhizome of Atractylodes lancea, was carried out by Aspergillus niger and Aspergillus cellulosae IFO 4040. Compound 1 was easily converted to compounds 2-9 by A. niger, and compounds 10 and 11 by A. cellulosae, respectively. Their stereostructures were established by a combination of high-resolution NMR spectral analysis, X-ray crystallographic analysis, and chemical reactions such as epoxydation. PMID- 10361703 TI - Production of pectic enzymes in yeasts. AB - When grown in the appropriate medium, several yeast species produce pectinases able to degrade pectic substances. It is mainly exocellular endopolygalacturonases that break pectins or pectate down by hydrolysis of alpha 1,4-glycosidic linkages in a random way. Biochemical characterisation of these enzymes has shown that they have an optimal pH in the acidic region and an optimal temperature between 40 and 55 degrees C. Their production by yeasts is a constitutive feature and is repressed by the glucose concentration and aeration. Pectic substances and their hydrolysis products are used as carbon sources by a limited number of yeasts and hence these enzymes must be involved in the colonisation of different parts of plants, including fruits. The first yeast pectic enzyme (encoded by the PSE3 gene) was cloned from Tichosporon penicillatum. Recently, a polygalacturonase-encoding gene from Saccharomyces cerevisiae has been cloned and overexpressed in several strains and the gene for an extracellular endopolygalacturonase from Kluyveromyces marxianus has also been described. Taking all the results together, the idea is now emerging that this type of yeast enzyme could offer an alternative to fungal enzymes for industrial applications. PMID- 10361704 TI - Plasmids of corynebacteria. AB - Corynebacteria are pleomorphic, asporogenous, Gram-positive bacteria. Included in this group are nonpathogenic soil corynebacteria, which are widely used for the industrial production of amino acids and detergents, and in biotransformation of steroids. Other members of this group are plant and animal pathogens. This review summarizes the current information available about the plasmids of corynebacteria. The emphasis is mainly on the small plasmids, which have been used for construction of vectors for expression of genes in these bacteria. Moreover, considerable information is now available on their nucleotide sequence, gene organization and modes of replication, which would make it possible to further manipulate these plasmids. Other plasmid properties, such as incompatibility and host range, are also discussed. Finally, use of these plasmids as cloning vectors for the expression of heterologous proteins using corynebacteria as hosts is also summarized to highlight the potential of these bacteria as hosts for recombinant DNA. PMID- 10361705 TI - Development of a transposon mutagenesis system for Mycobacterium avium subsp. paratuberculosis. AB - Mycobacterium avium subspecies paratuberculosis, a slow-growing Mycobacterium, is the causative agent of Johne's disease. Although M. paratuberculosis is difficult to manipulate genetically, our laboratory has recently demonstrated the ability to introduce DNA into these bacteria by transformation and phage infection. In the current study we develop the first transposon mutagenesis system for M. paratuberculosis using the conditionally replicating mycobacteriophage phAE94 to introduce the mycobacterial transposon Tn5367. Southern blotting and sequence analysis demonstrated that the transposon insertion sites are distributed relatively randomly throughout the M. paratuberculosis genome. We constructed a comprehensive bank of 5620 insertion mutants using this transposon. The transposition frequency obtained using this delivery system was 1.0 x 10(-6) transposition events per recipient cell. Auxotrophic mutants were observed in this library at a frequency of 0.3%. PMID- 10361706 TI - Characterisation of a chloramphenicol acetyltransferase determinant found in the chromosome of Pseudomonas aeruginosa. AB - The open reading frame (ORF) in the Pseudomonas aeruginosa chromosome, whose product resembles the chloramphenicol acetyltransferases (CAT) belonging to the CATB family, was cloned and shown to confer resistance to chloramphenicol (Cm) in Escherichia coli. The determinant was therefore named catB7 and the corresponding protein CATB7. When the copy number and expression signals were identical, the catB7 gene conferred resistance to Cm at a level slightly lower than those of three other catB genes. CATB7 resembles other CATBs in that it acetylates Cm but not 1-acetoxy-Cm. For CATB7, the K(m) values for acetyl-CoA and Cm were 5.0-5.4 fold higher than the corresponding values for each of the three other CATB proteins (CATB1, CATB3 and CATB5) examined and the Vmax was 5-6 fold lower. Using PCR, the catB7 gene was found in all six P. aeruginosa strains examined but not in any other species of pseudomonad tested. Weak CAT activity was detected in crude cell extracts from five of the six P. aeruginosa strains. However, this activity did not correlate with the Cm susceptibility of the strains, indicating that catB7 is not likely to be the major determinant of intrinsic Cm resistance in P. aeruginosa. PMID- 10361707 TI - The respiratory responses to cyanide of a cyanide-resistant Klebsiella oxytoca bacterial strain. AB - The respiratory system of a cyanide-resistant Klebsiella oxytoca was analyzed by monitoring the changes in the cytochrome contents in response to various inhibitors in the presence of various concentrations of cyanide. The cells grown in the medium without cyanide (KCN) have two terminal oxidases, cytochrome d (Ki = 10(-5) M KCN) and o (Ki = 10(-3) M KCN). When cells were grown on medium with 1 mM KCN, the expression of both b-type cytochrome and cytochrome d in the plasma membranes of the cell decreased by more than 50%, while cytochrome o increased by 70%, as compared with the cells grown in the absence of KCN. Two terminal oxidases with Ki values of about 10(-3) M and 1.7 x 10(-2) M KCN were observed in the plasma membrane fractions of the cells growing on KCN enriched medium. 2-n Heptyl-4-hydroxyquinoline-N-oxide markedly inhibited the oxidation of NADH by the plasma membranes from the cells grown in the medium without KCN, but not in those plasma membranes from KCN-grown cells. The NADH oxidases in plasma membranes of K. oxytoca grown with and without KCN were equally sensitive to UV irradiation. Adding freshly isolated quinone to the UV-damaged plasma membranes restored the NADH oxidase activity from both types of plasma membranes. From these results, we propose the presence of a non-heme type of terminal oxidase to account for the KCN resistance in K. oxytoca. PMID- 10361708 TI - Several highly divergent histone H3 genes are present in the hypotrichous ciliate Stylonychia lemnae. AB - In the protozoan Stylonychia lemnae 10 different histone H3 genes were discovered by polymerase chain reaction (PCR) amplification and sequence analysis. One of them is interrupted by a short intron sequence. These genes code for nine divergent histone H3 proteins. The genetic distances between some of these variants are very high. Most of the substitutions, as well as insertions/deletions, were found in the amino-terminal region. One variant shows an extremely elongated and altered N-terminus, which did not allow an unambiguous alignment with other histone H3 variants in this region. Hybridization experiments using the different H3 genes as probes indicate that even more histone H3 variants must exist in this species. PMID- 10361709 TI - Genome size and macrorestriction map of Xanthomonas campestris pv. glycines YR32 chromosome. AB - Xanthomonas campestris is an important plant pathogenic bacterium which causes severe diseases in a wide variety of plant species. We have generated a macrorestriction map of the X. campestris (axonopodis) pv. glycines chromosome employing pulsed-field gel electrophoresis (PFGE). Restriction endonucleases PacI (5'-TTAATTAA), PmeI (5'-GTTTAAAC) and SwaI (5'-ATTTAAAT) digested the chromosomal DNA into three, five, and five fragments, respectively. In addition, intron encoded restriction endonuclease I-CeuI was employed to locate the position of the 23S rRNA genes (rrlA and rrlB). All of the generated restriction fragments were aligned along the chromosome using multiple restriction enzyme digestion and two-dimensional PFGE (2-D PFGE) in conjunction with Southern hybridization analysis. This physical map construction has revealed a single circular chromosome with a size of approximately 5 Mb. Two rRNA genes were localized on the chromosome map. Several genes involved in pathogenesis (xpsD, opsX, and pat) as well as genes involved in the biosynthesis of xanthan gum (xanAB, rfbCDAB) were also localized. PMID- 10361710 TI - The expression of the flagellum of Legionella pneumophila is modulated by different environmental factors. AB - Legionella pneumophila, the aetiologic agent of legionnaires' disease, contains a single, monopolar flagellum which is composed of one major subunit, the FlaA protein. Expression studies using a reporter gene fusion of the flaA promoter with the luxAB gene revealed that the flaA expression is not only temperature regulated but is also influenced by the growth phase, the viscosity and the osmolarity of the medium, and by amino acids. PMID- 10361711 TI - The Mycobacterium marinum G13 promoter is a strong sigma 70-like promoter that is expressed in Escherichia coli and mycobacteria species. AB - A Mycobacterium marinum promoter, designated G13, was isolated from a promoter trap library as a constitutive producer of the mutant green fluorescent protein. Sequence analysis, primer extension analysis, and computer promoter prediction analysis indicate that the G13 promoter is very similar to Escherichia coli consensus sigma 70 promoters. Expression of the green fluorescent protein from the G13 promoter in M. marinum is, however, up to 40 times higher than that seen from the mycobacterial hsp60 promoter during exponential growth. Further, expression from this promoter does not appear to affect the growth of the organism in culture media or in macrophages. The strong expression of the G13 promoter allows it to be developed as a useful molecular tool for high level expression of markers in vitro. PMID- 10361713 TI - A new double-stranded RNA mycovirus from Botrytis cinerea. AB - A simple double-stranded RNA mycovirus was detected in a wild-type Botrytis cinerea 55k strain. The virus was located in the fungus cytoplasm as free particles of approximately 28 nm in diameter. The mycovirus possesses a single double-stranded genome segment of 1.8 kilobase pairs (kbp) encapsidated within an isometric protein coat whose main structural component is a polypeptide of 68 kDa. Cells infected with this virus showed an important degree of cellular degeneration. PMID- 10361712 TI - Differential effect of Cryptococcus neoformans on the production of IL-12p40 and IL-10 by murine macrophages stimulated with lipopolysaccharide and gamma interferon. AB - In the present study, we examined the in vitro effect of Cryptococcus neoformans on the production of interleukin-12 (IL-12) and IL-10 by murine macrophages. At a dose of 1 x 10(5), 1 x 10(6) or 1 x 10(7) ml-1, a highly virulent strain of C. neoformans (strain YC-11) suppressed the production of IL-12p40 by a murine macrophage cell line, J774.1 stimulated with lipopolysaccharide (LPS) and interferon (IFN)-gamma, while the production of IL-10 was not inhibited, but rather slightly augmented. The suppression of IL-12p40 production did not change by neutralizing anti-IL-10 mAb. A direct contact of C. neoformans with macrophages was largely involved in this inhibitory effect, since placement of a 0.45 micron pore membrane between the organism and macrophages prevented such effect. On the other hand, the culture supernatant of YC-11 did not inhibit macrophage IL-12p40 production when used at a lower dose, which contained an equivalent amount of capsular polysaccharide to that in the supernatant of YC-11 cultured at 1 x 10(5) or 1 x 10(6) ml-1, although it showed a small suppression at higher doses. Our results suggest that C. neoformans may suppress the induction of Th1 responses by inhibiting macrophage IL-12 production predominantly through a direct contact-dependent mechanism and to a lesser extent by a certain soluble factor(s) released from this microorganism. PMID- 10361714 TI - Molecular cloning of an immunogenic and acid-induced isocitrate dehydrogenase gene from Coxiella burnetii. AB - The Coxiella burnetii icd gene encoding an immunogenic dimeric NADP(+)-dependent isocitrate dehydrogenase (IDH) was cloned by screening a C. burnetii genomic library with a human positive serum and sequenced. The predicted gene product consists of 427 amino acids (M(r) = 46,600) and showed high identity to the IDHs of Escherichia coli (74%), Salmonella enterica (73%) and IDH-I of Vibrio sp. (71%). The cloned gene complemented an icd-defective E. coli mutant producing a recombinant IDH that had the same biochemical properties as the enzyme from purified C. burnetii. Unlike the homologs from other bacteria, the cloned enzyme was expressed to the highest level in low pH conditions. This distinct property of the cloned IDH suggests that C. burnetii icd gene may have a role in the adaptation of the organism to the harsh acidic environment of the eucaryotic phagolysosomes. PMID- 10361715 TI - Colony formation by Helicobacter pylori after long-term incubation under anaerobic conditions. AB - To evaluate the viability of Helicobacter pylori cultured under anaerobic conditions, H. pylori strain TK1029 was grown on blood agar in a microaerophilic environment at 37 degrees C for 4 days, and subsequently cultured under anaerobic conditions for 1 to 35 days. Colony formation by bacteria on blood agar plates cultured under anaerobic conditions was observed only for up to 4 days of microaerophilic incubation. By Gram staining, the morphological form of the bacteria was shown to be predominantly coccoid. However, bacteria cultured under anaerobic conditions for 15 to 35 days formed colonies on blood agar after pre incubation of bacteria with PBS, but not without pre-incubation. These results suggest that H. pylori survives long-term culture under anaerobic conditions and that both pre-incubation in non-nutrient solution and high density of bacterial concentration might be important for recovery of H. pylori cultured for a prolonged time under anaerobic conditions. PMID- 10361716 TI - Construction and immunologic evaluation of a Porphyromonas gingivalis subsequence peptide fused to hepatitis B virus core antigen. AB - Several proteins of Porphyromonas gingivalis contain multiple copies of a 47 amino acid conserved repeated sequence. A fusion protein was constructed in which the P. gingivalis peptide was fused to the carboxy terminus of the hepatitis B core protein. This fusion protein was expressed in Escherichia coli, purified, and used to vaccinate mice that were later challenged with P. gingivalis W83 using the mouse abscess model. Although the mice were not protected against bacterial challenge, Western blot analysis showed that sera from the mice and from rabbits immunized with the fusion protein reacted with a number of vesicle proteins from P. gingivalis W83. These data suggested that this peptide is recognized by the host's immune system but that the antibodies are not protective. PMID- 10361717 TI - Degradation of 2,4-dichlorophenol and pentachlorophenol by two brown rot fungi. AB - Wheat straw cultures of the brown rot fungi Gloeophyllum striatum and G. trabeum degraded 2,4-dichlorophenol and pentachorophenol. Up to 54% and 27% 14CO2, respectively, were liberated from uniformly 14C-labeled substrates within 6 weeks. Under identical conditions Trametes versicolor, a typical white rot species employed as reference, evolved up to 42% and 43% 14CO2 and expressed high activities of laccase, manganese peroxidase, and manganese-independent peroxidase. No such activity could be detected in straw or liquid cultures of Gloeophyllum. Moreover, G. striatum degraded both chlorophenols most efficiently under non-cometabolic conditions, i.e. on a defined mineral medium lacking sources of carbon, nitrogen and phosphate. PMID- 10361718 TI - Endogenous cytokines during a lethal infection with Listeria monocytogenes in mice. AB - It has been demonstrated that endogenous cytokines including gamma interferon (IFN-gamma), tumour necrosis factor-alpha (TNF-alpha), and interleukin-6 (IL-6) play protective roles but that IL-4 and IL-10 play detrimental roles in nonlethal Listeria monocytogenes infection in mice. In this paper, we studied the roles of endogenous cytokines in a lethal infection with L. monocytogenes in mice. TNF alpha and IL-6 titres in the bloodstreams, spleens and livers paralleled bacterial numbers in the organs, and both these cytokines and the bacterial numbers peaked just before the mice died. The high titres of TNF-alpha notably detected in the circulation in lethal infection were different from those in nonlethal infection. The maximum production of IFN-gamma was observed before the peaks of TNF-alpha and IL-6, and IFN-gamma almost disappeared from the bloodstreams and organs just before the mice died. No notable difference of IFN gamma titres between lethal infection and nonlethal infection in the specimens obtained from mice was observed. IL-10 was also detected in the bloodstreams earlier than the peaks of TNF-alpha and IL-6 during lethal infection, while IL-4 was never detected in the sera. The administration of monoclonal antibodies (mAbs) against TNF-alpha, IFN-gamma, IL-6, IL-4 or IL-10 failed to rescue mice from lethal L. monocytogenes infection, whereas anti-TNF-alpha mAb and anti-IFN gamma mAb prevented mice from lethality by high-dose endotoxin shock. These results suggest that lethality in L. monocytogenes infection might not be determined solely by these cytokines. PMID- 10361719 TI - Candida albicans exoglucanase as a reporter gene in Schizosaccharomyces pombe. AB - The Candida albicans XOG1 gene, previously shown to be a good reporter gene in Saccharomyces cerevisiae and C. albicans, was tested in Schizosaccharomyces pombe. Unlike the budding yeast, S. pombe does not produce exoglucanase activity and hence this system would be applicable to any given strain of this organism. The XOG1 gene was located under the control of the nmt1 promoter and its functionality could be demonstrated even at high temperatures (37 degrees C). The exoglucanase activity can be measured both in vivo and in vitro by either a simple biochemical reaction (on cells or media) or by flow cytometry, because the cells remain viable after the assay. PMID- 10361720 TI - Development of a serum-free medium for the production of erythropoietin by suspension culture of recombinant Chinese hamster ovary cells using a statistical design. AB - In order to develop a serum-free (SF) medium for the production of erythropoietin (EPO) by suspension culture of recombinant Chinese hamster ovary (rCHO) cells, a statistical optimization approach based on a Plackett-Burman design was adopted. A basal medium was prepared by supplementing Iscove's modified Dulbecco's medium (IMDM) with Fe(NO3)3.9H2O, CuCl2 and ZnSO4.7H2O which are generally contained in SF medium formulations. Insulin, transferrin and ethanolamine were also supplemented to the basal medium to determine their optimal concentrations. From this statistical analysis, glutamate, serine, methionine, phosphatidylcholine, hydrocortisone and pluronic F68 were identified as positive determinants for cell growth. The SF medium was formulated by supplementing the basal medium with components showing positive effects on cell growth in suspension culture. An EPO titer in this optimized SF medium was 79% of that in IMDM supplemented with 5% dialyzed fetal bovine serum (dFBS). Furthermore, the in vitro and in vivo biological activities of EPO produced in the SF medium were comparable to those produced in the serum-supplemented medium. Taken together, the results obtained here show that a Plackett-Burman design facilitates the development of SF media for the production of EPO by suspension culture of rCHO cells. PMID- 10361721 TI - Detection of the changing substrate requirements of cultured animal cells by stoichiometric growth equations validated by enthalpy balances. AB - As part of an overall aim to base the feeding of substrates to cultured animal cells on their actual metabolic needs, we have developed a stoichiometric approach centred on the macronutrients in the medium. Heat flux records the overall metabolic activity and therefore was the sensitive indicator of changing metabolic requirement. Analyses were made of the experimental measurements on two engineered cell lines in batch culture, the 2C11-12 macrophage hybridoma cell capable of the respiratory burst and the CHO320 constitutively producing human interferon-gamma. The crux was to construct simplified stoichiometric equations for the growth reactions to represent metabolic activity as it changed with time. Beforehand, it was essential to select the appropriate components for the equations. The choice was then justified by constructing enthalpy balances in which the ratio of heat flux to enthalpy flux must be close to unity for validation. By combining the stoichiometric approach with heat flow measurements, it was shown both theoretically and experimentally that the set of stoichiometric coefficients constituting a validated growth equation has a one-to-one corresponding relationship to the metabolic activity of the average cell population. Thus, a strategy was established for feeding the cells at any one time with the correct ratio of the major substrates, glucose and glutamine, in response to metabolic requirements that change with time. PMID- 10361722 TI - Generation of a new protein purification matrix by loading ceramic hydroxyapatite with metal ions--demonstration with poly-histidine tagged green fluorescent protein. AB - The gene encoding the green fluorescent protein (GFP) from the jellyfish Aequorea victoria, was inserted under transcriptional control of the polyhedrin promoter of the Autographa californica nuclear polyhedrosis virus and expressed in the Spodoptera frugiperda insect cell line Sf9 during viral infection. The baculovirus transfervector pBlueBacHisB was used for constructing the recombinant baculovirus, so that the green fluorescent protein could be tagged with a poly histidine tail. This fusion protein was utilized as a marker for evaluating the properties of metal ion loaded ceramic hydroxyapatite as a matrix in protein purification. Ceramic hydroxyapatite loaded with Zn(II) was the best choice for purifying this poly-histidine tagged GFP, followed by Fe(III) of the metal ions tested. Ni(II) that is superior especially in many poly-histidine purification systems did not, when loaded to hydroxyapatite, have binding properties comparable to Zn(II) or Fe(III). Elution of poly-histidine tagged GFP was best performed with phosphate buffers or EDTA that could compete with the phosphate molecules in hydroxyapatite or complexly bind the metal ions, respectively. PMID- 10361723 TI - Glucose 1- and 2-oxidases from fungal strains: isolation and production of monoclonal antibodies. AB - Monoclonal antibodies (Mabs) against purified glucose 2-oxidase (EC 1.1.3.10) from Coriolus versicolor were raised by hybridoma technology using Sp2/0 myeloma cells as a fusion partner. Hybrid growth was observed in 42% of culture wells and 30% of these (i.e. 30 culture wells) contained anti-glucose 2-oxidase activity. Three positive wells containing hybrid cell lines were selected and cloned twice by the limiting dilution method and two hybridoma clones (E1A5 and E1A6) secreting Mabs were selected at random for purification and characterisation purposes. Both cell lines secreted Mabs of IgM class which were purified by gel filtration chromatography on a Sephacryl S-200 column with a final recovery of 80% and a purification factor of 16. The purified preparations were apparently homogeneous on native PAGE running with a M(r) of 950 kDa. Mabs were highly specific for glucose 2-oxidase as determined by Western blotting. These Mabs also crossreacted with glucose 1- and 2-oxidases from other fungal sources (Phanerochaeta chrysosporium, Penicillium amagasakiense and Aspergillus niger) as determined by Western blotting and by ELISA. Both glucose 1- and 2-oxidases from C. versicolor, P. chrysosporium, P. amagasakiense and A. niger were purified by hydrophobic interaction chromatography on Sepharose 4B-triazine dye with a recovery of enzyme activity in the range 85-92%. Purified preparations of glucose oxidases from fungal strains were apparently homogeneous on native PAGE. Glucose 2-oxidases were more hydrophobic than glucose 1-oxidases as determined by their chomatographic behaviour on Sepharose 4B-Cibacron Red G-E which could be used to study their roles in lignin biodegradation. PMID- 10361724 TI - Effects of R-(+)-limonene on submerged cultures of the terpene transforming basidiomycete Pleurotus sapidus. AB - The biotransformation of limonene by the basidiomycete Pleurotus sapidus yielded cis/trans-carveol and carvone as the main products. The transformation period was extended from 4 days after direct addition to 12 days by gas phase addition of the substrate. After 2 days of transformation, 97% of the substrate had accumulated in the mycelium, while only 3% were present in the culture medium. Substrate toxicity led to a decrease of dry matter. Adaptation of the precultures with small amounts of substrate doubled the concentration of carveol and increased the concentration of carvone by a factor of 3-4. Total product concentrations of > 100 mg l-1 were reached. PMID- 10361725 TI - A recombinant Fab neutralizes dengue virus in vitro. AB - A recombinant Fab that recognizes a neutralizing epitope located in the (296-400) region of protein E of dengue virus was obtained from cloned hybridoma cells secreting the mouse monoclonal antibody (mAb) 4E11. The Fd and light chain antibody genes were amplified by polymerase chain reaction, cloned into the phagemid vector pMad, expressed in bacteria to produce Fab fragments and sequenced. The mAb 4E11, in particular its light chain complementary-determining regions, shared homologies with two other anti-viral mAbs. The affinity of the parental mAb and the cloned Fab to the MalE-E(296-400) fusion protein were shown to be of the same magnitude, i.e. nanomolar. Fab 4E11 neutralization capacity was found between 8 and 4-times or less lower than that of mAb 4E11, depending on serotypes, thus the Fab could have a smaller antiviral activity than the mAb in vitro. PMID- 10361726 TI - Rapid structural characterisation of a murine monoclonal IgA alpha chain: heterogeneity in the oligosaccharide structures at a specific site in samples produced in different bioreactor systems. AB - A murine hybridoma cell line which secretes a monoclonal IgA antibody, directed against the LPS antigen of Vibrio cholerae, was grown in either a continuous stirred tank reactor, a fluidised bed reactor or a hollow fibre reactor. Different methods were used for the structural characterisation of the IgA alpha chains. A classical approach consisted of Edman sequencing and mass determination of peptides separated by reversed phase HPLC. Alternatively, peptides and glycopeptides from a tryptic digest of each alpha chain were identified directly by MALDI-TOF mass spectrometry. A detailed analysis of the oligosaccharide structures at an unique site on the alpha chain was made by labelling the oligosaccharides released by N-glycosidase F with 1-(p-methoxy)phenyl-3-methyl-5 pyrazolone. After separation by HPLC, mass measurements were made using matrix assisted laser desorption time of flight mass spectrometry before and after digestion with specific exoglycosidases. The primary structure of the alpha chain of IgA was not affected by different cell culture conditions; in contrast, significant variations could be detected in the pattern of N-linked oligosaccharide structures, most prominently in the degree of sialylation. The efficiency of the analytical techniques in providing quality control of the identity, integrity and consistency of the glycoprotein is shown and discussed. PMID- 10361727 TI - The isolation of strains of Saccharomyces cerevisiae showing altered plasmid stability characteristics by means of selective continuous culture. AB - A recombinant strain of Saccharomyces cerevisiae containing a plasmid-encoded lacZ gene from Escherichia coli was grown for 420 generations under selective conditions in glucose-limited continuous culture. A ura3-based auxotrophic system was used to apply selection in favour of plasmid-containing organisms. A similar strategy had previously proved successful at evolving clones of Bacillus subtilis, showing improved plasmid stability characteristics. In this study a series of clones were isolated which exhibited large variation in their ability to retain the recombinant plasmid. Clones showed both significantly increased and reduced capacity to maintain the recombinant plasmid. The probabilities of obtaining clones in either category were essentially equal so that selection was not seen to enrich for more stable clones. Periodic selection events appeared to exert a greater influence on the distribution of stability characteristics amongst clones than did the applied selective pressure. Alterations in plasmid retention characteristics could be associated with host or plasmid. The most stable clone isolated exhibited a approximately 30% improvement of its overall stability (sigma(N+)) and an 80% improvement in productivity, when compared to the parental strain CGpLG. This improved stability was associated with alterations in the plasmid genome. PMID- 10361728 TI - Changes during subclone development and ageing of human antibody-producing recombinant CHO cells. AB - Some of the problems encountered with human or human-mouse heterohybridomas, such as low growth rates and high serum requirements, have led to the increased use of recombinant cell lines for production of human antibodies. To evaluate the suitability of such alternative cell lines for the production of human antibodies we have analysed several subclones with differing specific production rates of a recombinant CHO cell line. Gene copy number and site of chromosomal integration for the light and heavy chain and the dhfr gene were determined by in-situ hybridisation. Specific mRNA content was analysed by Northern blot. In addition the intracellular content in light and heavy chain was measured by flow cytometry and the specific secretion rates were determined. The stability of gene expression was followed in the highest producing subclone for over a year. As previously seen in heterohybridoma cells a high expression rate of light chain is beneficial in speeding up secretion rates of whole antibody. When grown in the presence of G418 and methotrexate the amplified gene copies in the genome of recombinant CHO cells were stable over more than 100 passages. However, the expression of light chain, and with it the secretion rate, decreased with time. The low intracellular concentration of light chain resulted in accumulation of heavy chain in the endoplasmic reticulum due to retention by chaperones. The specific secretion rate decreased by 50% after 100 passages. When no G418 or methotrexate were present 75% of the gene copies were lost after 100 passages. PMID- 10361729 TI - Pharmacophore fingerprinting. 1. Application to QSAR and focused library design. AB - A new method of rapid pharmacophore fingerprinting (PharmPrint method) has been developed. A basis set of 10,549 three-point pharmacophores has been constructed by enumerating several distance ranges and pharmacophoric features. Software has been developed to assign pharmacophoric types to atoms in chemical structures, generate multiple conformations, and construct the binary fingerprint according to the pharmacophores that result. The fingerprint is used as a descriptor for developing a quantitative structure-activity relationship (QSAR) model using partial least squares. An example is given using sets of ligands for the estrogen receptor (ER). The result is compared with previously published results on the same data to show the superiority of a full 3D, conformationally flexible approach. The QSAR model can be readily interpreted in structural/chemical terms. Further examples are given using binary activity data and some of our novel in house compounds, which show the value of the model when crossing compound classes. PMID- 10361730 TI - Development of three-dimensional descriptors represented by tensors: free energy of hydration density tensor. AB - In order to describe the degree of interaction of a molecule with its environments by descriptors, several three-dimensional descriptors have been proposed. With the physical properties calculated around a molecule, scalar, vector, and tensor (zeroth, first, and second moments) of the physical properties were calculated and were used as descriptors for calculating the similarity index between the molecules. The tensors contain the information on the spatial distribution of those physical properties around the molecule. Hydration Free Energy Density (HFED) proposed by No et al. was used to calculate HFED tensor. The descriptors were used for the similarity index calculations between substituted benzenes and between lead compounds of HIV-1 protease inhibitors. The substituted benzenes are grouped according to the similarity indices. The grouping seems reasonable from the viewpoint of a chemical sense. The lead fragments of the HIV-1 protease inhibitors have a high similarity among themselves though their chemical formulas are not very similar, the lead fragments are diverse. Although the chemical formulas are diverse, the spatial distribution of the physical properties around the molecules is similar. The descriptors have high discriminating power in the similarity calculation between the molecules. PMID- 10361731 TI - Assembly of the paraflagellar rod and the flagellum attachment zone complex during the Trypanosoma brucei cell cycle. AB - Trypanosomes possess a single flagellum that is attached to their cell body via the flagellum attachment zone (FAZ). The FAZ is composed of two structures: a cytoplasmic filament complex and four microtubules situated next to it. There is a complex transmembrane crosslinking of this FAZ to the paraflagellar rod (PFR) and axoneme within the flagellum. We have partially purified the FAZ complex and have produced monoclonal antibodies both against the FAZ and the paraflagellar rod. The two antibodies against the FAZ (L3B2 and L6B3) recognise the cytoplasmic filament in immunofluorescence and in immunoelectron microscopy. On western blot, they detect a doublet of high molecular weight (M(r) 200,000). Two anti-PFR antibodies (L13D6 and L8C4) recognise the paraflagellar rod in immunofluorescence, but show a difference on Western blot: L13D6 recognises both major PFR proteins, whereas L8C4 is specific for only one of them. Using these new antibodies we have shown that although the growth of both cytoplasmic FAZ filament and external PFR are related, their growth initiates at different time points during the cell cycle and the two structures elongate at distinct rates. PMID- 10361732 TI - Encephalitozoon cuniculi: light and electron microscopic evidence for di-, tetra , and octosporous sporogony and a note on the molecular phylogeny of encephalitozoonidae. AB - We demonstrate, based on the light, electron microscopic, and immunofluorescence studies carried out on two isolates of Encephalitozoon cuniculi established in culture, that E. cuniculi exhibits di-, tri-, tetra- and octosporous sporogony. We therefore propose that the generic characters of Encephalitozoon should be amended to include tetra-sporous sporogony as generic features. Additionally, the molecular phylogenetic analysis indicates that E. cuniculi, E. hellem, and E. (Septata) intestinalis form a cohesive group. PMID- 10361733 TI - Critical analysis of eukaryotic phylogeny: a case study based on the HSP70 family. AB - Trichomonads, together with diplomonads and microsporidia, emerge at the base of the eukaryotic tree, on the basis of the small subunit rRNA phylogeny. However, phylogenies based on protein sequences such as tubulin are markedly different with these protists emerging much later. We have investigated 70 kDa heat-shock protein (HSP70), which could be a reliable phylogenetic marker. In eukaryotes, HSP70s are found in cytosol, endoplasmic reticulum, and organelles (mitochondria and chloroplasts). In Trichomonas vaginalis we identified nine different HSP70 encoding genes and sequenced three nearly complete cDNAs corresponding to cytosolic, endoplasmic reticulum, and mitochondrial-type HSP70. Phylogenies of eukaryotes were reconstructed using the classical methods while varying the number of species and characters considered. Almost all the undoubtedly monophyletic groups, defined by ultrastructural characters, were recovered. However, due to the long branch attraction phenomenon, the evolutionary rates were the main factor determining the position of species, even with the use of a close outgroup, which is an important advantage of HSP70 with respect to many other markers. Numerous variable sites are peculiar to Trichomonas and probably generated the artefactual placement of this species at the base of the eukaryotes or as the sister group of fast-evolving species. The inter-phyla relationships were not well supported and were sensitive to the reconstruction method, the number of species; and the quantity of information used. This lack of resolution could be explained by the very rapid diversification of eukaryotes, likely after the mitochondrial endosymbiosis. PMID- 10361734 TI - Rumen ciliates of white-tailed deer (Odocoileus virginianus), axis deer (Axis axis), sika deer (Cervus nippon) and fallow deer (Dama dama) from Texas. AB - Samples of rumen contents from 33 white-tailed deer (Odocoileus virginianus), 31 axis deer (Axis axis), 26 sika deer (Cervus nippon), and 25 fallow deer (Dama dama) were collected from four study areas in central Texas. The geometric mean concentration of total protozoa was 50.2 x 10(4) per ml, with no differences between species (P > 0.36). White-tailed deer had a higher percentage of Entodinium and lower percentage of Diplodiniinae (P < 0.01) than the other deer species, which were not different from each other. Occurrence of Epidinium, Isotricha, and Dasytricha was sporadic and did not differ among deer species. Numerous new host records of protozoan species were observed: white-tailed deer- four; axis deer--five; sika deer--five; fallow deer--four. This brings the total number of protozoan species identified in each deer species to: white-tailed- eight; axis--12; sika--15; fallow--16. For all species combined, protozoan concentration were 7.5 to 11-fold higher (P < 0.01) from Area 4, which differed from the other three areas by having a stream that allowed deer to have free access to water. Criteria used for identification of medium-size Eudiplodinium species were evaluated. PMID- 10361735 TI - Affinity-purification of concanavalin A-binding ciliary glycoconjugates of starved and feeding Tetrahymena thermophila. AB - Development of mating competency in Tetrahymena thermophila requires starvation for at least 70 min in low ionic strength buffer. Pair formation between conjugating cells is blocked at early stages by the lectin Concanavalin A (Con A). To investigate the role of Con A-binding proteins in this induced cellular change and pairing, and to confirm and extend an earlier study from our laboratory, a method was developed for preparation of Con A-binding proteins from ciliary membrane-rich fractions of T. thermophila. Con A-binding ciliary proteins were prepared from non-starved and starved cells from two wild type strains and a mating mutant, RH179E1. Comparison of these proteins by SDS-PAGE revealed on overall reduction in number of wild-type bands after starvation. In particular, a major band at 28 kDa was present in non-starved cells and absent in starved cells. However, in the mating mutant, no change in banding profile was seen after starvation: the 28 kDa band was present in both non-starved and starved cells. This, Con A-binding ciliary membrane proteins undergo a major change during starvation-induced development of mating competency in wild-type T. thermophila. In contrast, the mutant differed from wild-type in overall composition of its ciliary Con A-binding glycoproteins and in the response of these proteins to starvation, suggesting that it may be deficient in its ability to be initiated by starvation. Our results are consistent with the hypothesis that a change affecting ciliary membrane Con A-binding proteins is essential for the cellular response to mating signals. PMID- 10361736 TI - New axonemal dynein heavy chains from Tetrahymena thermophila. AB - Two dyneins can be extracted from Tetrahymena ciliary axonemes. The 22S dynein contains three heavy chains (HC), sediments at 22S in a sucrose gradient, and makes up the outer arms. The 14S dynein contains two to six HCs, sediments at 14S, and is thought to contribute to formation of the inner arms. We have identified two large proteins that are extracted from Tetrahymena axonemes with high salt and that sediment together at approximately 18S. The two large proteins cleave when subjected to UV light in the presence of ATP and vanadate, suggesting both proteins are dynein HC. Antibodies against one of the 18S HCs do not recognize 22S dynein HCs. Antibodies to 22S dynein HC do not bind appreciably to 18S dynein photocleavage fragments. Taken together, these results indicate that the large proteins that sediment at 18S are axonemal dynein heavy chains. PMID- 10361737 TI - Identification of Trypanosoma cruzi zymodemes by kinetoplast DNA probes. AB - Analysis of zymograms of extracts of Trypanosoma cruzi isolated from different hosts in Argentina allowed characterization of 12 zymodemes or "isozymic strains," only six of which were found in human patients. Two of these six zymodemes (Z1 and Z12) were widely distributed and found in more than 80% of human patients. These two "major natural clones" differed significantly in pathogenic activity. Because the groupings obtained by studying enzymes and kinetoplast DNA (kDNA) were similar, it is possible to identify the zymodeme by analyzing kDNA. A 290-bp fragment was amplified by PCR using primers for the sequences flanking the hypervariable regions of kDNA minicircles. Labeled probes for this fragment, prepared from Z1 and Z12 reference stocks, hybridized specifically with PCR-amplified kDNA from parasite stocks, allowing identification of zymodemes. PMID- 10361738 TI - Cloning and expression of a cDNA encoding a Vorticella convallaria spasmin: an EF hand calcium-binding protein. AB - The stalked, ciliated protozoan Vorticella convallaria possesses a highly contractile cytoskeleton consisting of spasmonemes and myonemes. The major component of these contractile organelles is the calcium-binding protein(s) called spasmin. Cloning and characterization of spasmin would help elucidate this contractile system. Therefore, enriched spasmoneme protein preparations from these contractile stalks were used to produce a monoclonal antibody to spasmin. A monoclonal antibody, 1F5, was obtained that immunolocalized specifically to the spasmonemes and the myonemes and recognized a 20-kD calcium-binding protein in spasmoneme protein preparations. A putative spasmin cDNA was obtained from a V. convallaria cDNA library and the derived amino acid sequence of this cDNA revealed an acidic, 20-kD protein with calcium-binding helix-loop-helix domains. The physical properties of the putative spasmin were assessed by characterization of a recombinantly-produced spasmin protein. The recombinant spasmin protein was shown to bind calcium using calcium gel-shift assays and was recognized by the anti-spasmin antibody. Therefore, a V. convallaria spasmin was cloned and shown to be a member of the EF-hand superfamily of calcium-binding proteins. PMID- 10361739 TI - The phylogenetic position of Amoebophrya sp. infecting Gymnodinium sanguineum. AB - The small-subunit rRNA sequence of a species of Amoebophrya infecting Gymnodinium sanguineum in Chesapeake Bay was obtained and compared to the small subunit rRNA sequences of other protists. Phylogenetic trees constructed with the new sequence place Amoebophrya between the remaining dinoflagellates and other protists. PMID- 10361740 TI - Characterization of the Euplotes crassus macronuclear rDNA and its potential as a DNA transformation vehicle. AB - We have cloned the macronuclear linear DNA molecule carrying the ribosomal RNA genes from the ciliated protozoan Euplotes crassus. DNA sequence analysis was carried out to locate coding regions and to determine whether sequences that have been mutated to confer antibiotic resistance are conserved in the E. crassus genes. The beginning and end of the primary transcript were mapped. In order to determine whether conserved sequences that might serve as replication origins were present, the 5' and 3' non-coding sequences from E. crassus were compared to the corresponding sequences from the macronuclear linear rDNA molecules from the following euplotid species: Euplotes vannus, Euplotes minuta, Euplotes raikovii and Euplotes rariseta. A DNA transformation construct was made by generating a putative anisomycin resistant mutation along with a mutation generating a restriction site polymorphism. Microinjection of the construct into the developing macronucleus of mated cells resulted in exconjugant cell lines with increased resistance to anisomycin. The injected rDNA with the restriction site polymorphism is detectable in the anisomycin resistant cells and appears to represent a minor fraction of the rDNA. PMID- 10361741 TI - A molecular study of euglenoid phylogeny using small subunit rDNA. AB - The euglenoids are an ancient and extremely diverse lineage of eukaryotic flagellates with unclear relationships among taxa. Synapomorphies for the euglenoids include a surface pellicle and a closed mitosis with a series of separate sub-spindles. The taxonomy currently in use is inconsistent with the available data and needs revision. Most euglenoid phylogenies are largely intuitive reconstructions based on a limited number of morphological characters. Therefore, we have added molecular characters from the Small Subunit (SSU) rDNA to generate an overall phylogenetic framework for the euglenoids. SSU rDNA sequences from photosynthetic, osmotrophic, and phagotrophic euglenoids were aligned based on secondary structure. Phylogenetic analysis using the conserved areas of the sequence was performed using parsimony, maximum likelihood, and distance methods. Trees derived using different criteria are in agreement. The euglenoids form a distinct monophyletic clade with phagotrophic members diverging prior to the phototrophic and osmotrophic members. Among photosynthetic members, the biflagellate form diverged prior to the uniflagellate form. Additionally, the genus Euglena appears to be paraphyletic, with osmotrophic taxa, such as Astasia and Khawkinea, diverging independently within the clade containing the photosynthetic genus Euglena. PMID- 10361742 TI - [Myringoplasty in children: a comparison with an adult population]. AB - The indications for myringoplasty in children has always been a controversial subject since many authors feel the high frequency of the phlogistic auricular processes in children constitute an unfavorable prognostic factor to success of the procedure. The authors present the results obtained in 23 patients under 17 years of age who had undergone myringoplasty for simple perforation of the tympanum. In cases of posterior and inferior perforations, surgery was performed using the underlay technique and a transmeatal approach; in all other cases the overlay technique was used with a retroauricular approach. An average 30 month follow-up (range 12-55 months) revealed new perforations in only 2 cases (9%). From the functional point of view the average air/bone conduction gap was reduced to 10 dB. As a control, the results were compared to those obtained in 150 patients over 16 years of age, again affected by simple perforation of the tympanic membrane and treated by myringoplasty using the same methods. In the adults, 22 new perforations were found (15%) while the functional results were analogous to those obtained in the children group. In this light, it can be asserted that myringoplasty can be considered a safe procedure to be used in children and it does not appear essential to wait until they have finished growing before performing this procedure. PMID- 10361743 TI - [Canalith repositioning maneuver: proposal of a new therapy for benign paroxysmal positional vertigo of the posterior semicircular canal]. AB - A new therapy is proposed for the treatment of Benign Paroxysmal Positional Vertigo (B.P.P.V.) of the Posterior Semicircular Canal (P.S.C.): the Canalith Repositioning Maneuver (C.R.M.). The need for a new maneuver to treat B.P.P.V. of the P.S.C. arises from the difficulties encountered in daily practice, under particular conditions (i.e. elderly, obese, traumatized patients and in the presence of rachis pain, etc.), to perform the most common rehabilitative techniques such as the Semont Maneuver and Epley's Canalith Repositioning Procedure (C.R.P.). The results achieved using this new technique on a group of 47 consecutive out-patients are presented and compared to those achieved using the Semont Maneuver in an analogous group of 23 patients. C.R.M. and the Semont Maneuver were performed once per treatment session and all patients were checked every 3 days until the symptoms of B.P.P.V. disappeared. Thereafter they were invited to return for check-up if signs of vertigo returned (follow-up 6-25 months). The two techniques proved equally effective: 87.5% of the cases were resolved with C.R.M., 82.6% with the Semont Maneuver. However, the C.R.M. provided other advantages as it resolved the problem immediately (i.e. in a single session) in 81% of the cases vs. 68.4% for the Semont Maneuver. In view of the Canalith theory, the action mechanism envisaged for all three maneuvers- C.R.M., the Semont Maneuver and the Epley C.R.P.--can be explained assuming that the canalith passes from the ampullar to the non ampullar branch of the (P.S.C.) passing finally through the Common Duct and into the Utricle. C.R.M. is a specific treatment for the B.P.P.V. of the P.S.C. and is simple to perform, well tolerated and quite effective. It is indicated in all cases of B.P.P.V. of the P.S.C. both as initial treatment and as alternative to other treatment methods which have proved ineffective or difficult to perform. Indeed, in therapy it is best to be quite skilled in more than one technique, availing oneself of a full range possibilities; in this way the cure can be tailored to the patient in each individual case and not vice versa. PMID- 10361744 TI - [The treatment of choanal atresia]. AB - Choanal atresia occurs in approximately 1 out of every 8000 live births. In about 60% of these cases it is unilateral and is often associated with other major craniofacial anomalies or visceral malformations such as the so-called CHARGE syndrome (i.e. Choloboma, Heart defects, choanal Atresia, Retarded growth and development, Genital anomaly, and Ear defect with deafness) first described by Pagon in 1981. In newborns the clinical manifestations of choanal atresia consist of respiratory distress strictly related to the entity of nasal obstruction. Total bilateral congenial choanal atresia must, therefore, be considered a surgical emergency since nasal obstruction impedes the suction mechanism and hence normal feeding in the newborn. For this reason, prompt surgery is mandatory. The surgical approach employed in choanal atresia consists of both the trans-nasal and trans-palatal approaches. Between 1985 and 1997 31 patients with choanal atresia were treated in our department (16 males, 15 females; age range 2 days-5.5 years; mean 11.4 months). In 25 cases (80.6%) the malformation was bilateral. Associated anomalies were seen in 8 patients (25.8% of cases). All of these patients were treated using the trans-nasal approach, under general anesthesia, and endoscopic control. In 27 cases a trocar was employed to perforate the bony atresia and a stent was then positioned. Three patients underwent contact-laser resection of the stenosis without requiring any further stent and without any complications. None of these patients developed total recurrences, although after varying amounts of time, in 8 of the 27 patients operated using the trocar (29.6% of cases) a partial reduction of the airway occurred, the substenosis requiring dilatation with progressive Hegar dilators. In the remaining cases contact-laser therapy was associated with dilation. Three of the patients were treated by laser-therapy alone and none developed a recurrence. One patient dropped out of the follow-up for other serious malformations. PMID- 10361745 TI - [Surgical reconstruction of the external nose: personal experience]. AB - Because of the ensuring malformations, most often vascular in nature, malignant tumors of the nasal pyramid often require partial or total exeresis of the nasal structures. These tumors are most commonly basal cell carcinoma (85-90%) and squamous cell carcinoma (7-8%). A recent epidemiological survey performed in Italy showed that, in this country, such carcinomas account for approximately 13% of all malignant cutaneous neoplasms of the head and neck region. This work presents the author's personal experience in 8 cases of partial, subtotal and total reconstruction of the nose, performed over 5 years of clinical activity. The cases include 8 patients (4 males and 4 females, age range 48-95 years, mean age 77.3 years): 7 cases of malignant cutaneous nasal tumors (5 basal cell, 1 squamous cell and 1 basal-squamous cell carcinoma) and 1 case of iatrogeneous perforation through the entire thickness of the right nasal wall, approximately 7 8 mm in diameter, the outcome of sclerotizing treatments initiated 30 years before for angioma. On the whole, including the case of iatrogeneous nasal perforation the following procedures were used: 7 frontal fascio-cutaneous flaps and 1 nasal-genienic flap associated with a median of upper lip muscle. As regards the frontal flaps, in 5 cases a variation was used where the inset base was set more medially and with dimensions different from up-and-down Converse flaps. In 2 other cases a median frontal flap was used. Except for one case of partial necrosis of the flap, healed by subsequent surgery, complications were not particularly important from the clinical point of view and occurred in the broadest exeresis, where reconstruction of the structures adjacent to the nose and the mucosa lining proved necessary. On the whole, considering the initial stages of the lesions and the often advanced age of the patients, the surgical treatment gave good results from the oncological, aesthetic and functional points of view. PMID- 10361746 TI - [Wegener's and Stewart's granulomatosis: a case report of Stewart's granulomatosis]. AB - Wegener's and Stewart's diseases are two rare pathologies of unknown origin; both can cause disruptive facial lesions. Wegener's disease is a systemic pathology and generally involves the kidneys and lungs. Granulomatosis lesions are characterized by the necrotizing vasculitis involving the small vessels. From a diagnostic point of view the systemic features of Wegener's disease and the specific immunological findings (i.e. IgG autoantibodies vs. monocyte and neutrophil cytoplasm) make it possible to diagnose the disease precisely. Stewart's disease can be differentiated from Wegener's disease by the absence of any systemic lesions and the lack of necrotizing vasculitis. Pathogenesis of the disease is still unknown although immunohistochemical findings indicate that it is related to extranodal Tcell lymphomas. Stewart's disease is very aggressive with massive destruction of the midface tissues and prognosis is very poor (from a few months to 23 years). Surgery is generally ineffective in such disorders. The treatment for Wegener's disease includes the use of systemic steroids, immunosuppressive drugs and the sulfametoxazole-thrimethoprime association while radiotherapy associated with chemotherapy appears most effective in Stewart's disease. The authors describe a case of Stewart's disease prevalently involving the nasal cavities, ethmoid and paranasal sinuses. Diagnosis was made on the basis of immunohistochemical, histomorphological and immunological data. Treatment--based initially on systemic steroids with the association thrimethoprimsulfometoxazole--induced significant disease remission. Subsequent use of cytostatic drugs has made it possible to control the disease progression to date. PMID- 10361747 TI - [Septic thrombophlebitis of the internal jugular vein due to Fusobacterium necrophorum (Lemierre's syndrome): case report and review of literature]. AB - Lemierre's syndrome is an uncommon clinical entity. It is characterized by oropharyngeal infection followed by septic thrombophlebitis of the jugular vein with embolization to the lungs and other organs. It is usually due to Fusobacterium necrophorum. Although Lemierre's syndrome is rare, it is potentially fatal. It is important for clinicians to recognize and treat it appropriately. With prompt recognition, abscess drainage with possible ligation of the internal jugular vein and appropriate antibiotic coverage complete recovery can be achieved in most patients. PMID- 10361748 TI - [Basaloid-squamous cell carcinoma of the larynx: a new morphologic entity. Case report]. AB - Basaloid-Squamous Cell Carcinoma (BSCC) is considered a biologically aggressive variation of squamous cell carcinoma (SCC). Wain et al. first reported BSCC as an independent neoplasm in 1986. In the past, it was most likely misinterpreted as atypical SCC. To date approximately one hundred cases have been reported in the literature. The pathognomonic characteristic of BSCC is an intimate association between basaloid and squamous patterns: for this reason a small biopsy specimen may be insufficient for correct diagnosis. In such cases, knowledge of the biological behavior of these tumors (i.e. local aggressiveness and tendency to spread both regionally and at a distance) may prove useful. Prognosis and survival rates are certainly worse than in SCC. Within the upper aero-digestive tract, BSCC is most likely to arise in the supraglottic larynx, the pyriform sinus and the base of the tongue. Treatment should include surgery of the primary tumor and of the neck lymph nodes, followed by radiotherapy. The authors present a new case report indicating the typical feature of this tumor (aggressiveness, synchronous basaloid and squamous histology, supraglottic involvement. PMID- 10361749 TI - [Guidelines on interventional and diagnostic or/and therapeutic procedures applicable in day surgery in otolaryngology and cervical-facial surgery. Italian Society of Otorhinolaryngology and Cervico-Facial Surgery]. PMID- 10361750 TI - Treat tobacco dependence and "bend the trend". PMID- 10361751 TI - Strong bones in later life. PMID- 10361752 TI - Kala-azar epidemic in Varanasi district, India. AB - Reports at the Sir Sunder Lal Hospital, Banaras Hindu University, of a large number of kala-azar cases from one particular village in Varanasi district, Uttar Pradesh, led us to carry out an epidemiological study of the situation using standard techniques. The overall prevalence and case fatality of the disease were 12.9% and 10.5%, respectively. A history of fever and hepatosplenomegaly was noted for all the cases. The case definition was the presence of parasites in bone marrow or splenic aspirate smears. The disease was more prevalent among adults, but occurred also among children. However, there was no clear linear relationship between the prevalence of the disease and age group. Kala-azar occurred among males and females, and its prevalence did not correlate significantly with income. Since the disease vector continues to be present in the study area, the health authorities should take strong steps to control the disease. PMID- 10361753 TI - Seasonal diarrhoeal mortality among Mexican children. AB - The study investigated the effects on diarrhoeal deaths among under-5-year-old Mexican children of the following variables: season (summer or winter), region (north versus south), age group, and place of death. Examination of death certificates indicated that the distribution of deaths in 1989-90 was bimodal, with one peak during the winter and a more pronounced one during the summer. In 1993-94, however, the winter peak was higher than that in the summer (odds ratio (OR) = 2.04). These findings were due mostly to deaths among children aged 1-23 months (OR = 1.86). Diarrhoeal mortality was highest among children aged 6-11 months (OR = 2.23). During the winter, there was a significant increase in the number of deaths that occurred in medical care units and among children who had been seen by a physician before they died, but deaths occurring at home showed no seasonal variation. In the northern states, the reduction in diarrhoeal mortality was less in winter than in summer (OR = 2.62). In the southern states, the proportional reduction during the winter was similar to that in the summer. PMID- 10361754 TI - Effect of breastfeeding education on the feeding pattern and health of infants in their first 4 months in the Islamic Republic of Iran. AB - This quasi-experimental study was conducted in Shiraz, the Islamic Republic of Iran, on 120 pairs of mothers and infants in a maternity hospital that had a rooming-in programme. All 59 mothers in the study group received breastfeeding education, face-to-face, after delivery and during follow-up for 4 months in the mother and child health (MCH) centre or in their homes; the remaining 61 mothers comprised the control group. Exclusive breastfeeding rates were significantly higher in the study group (54%) than in the control group (6.5%), but 5% and 18% of infants, respectively, in the study and control groups had stopped breastfeeding by the age of 4 months. The mean number of days of diarrhoea experienced by infants in the study group were significantly lower (P < or = 0.004) than in the control group. At the end of 4 months, the mean weight and length of the infants were significantly higher (both P < 0.05) in the study group than in the control group. The findings indicate that rooming-in is very important for promoting exclusive breastfeeding and that there is a need for continuous breastfeeding education of mothers. PMID- 10361755 TI - True status of smear-positive pulmonary tuberculosis defaulters in Malawi. AB - The article reports the results of a study to determine the true outcome of 8 months of treatment received by smear-positive pulmonary tuberculosis (PTB) patients who had been registered as defaulters in the Queen Elizabeth Central Hospital (QECH) and Mlambe Mission Hospital (MMH), Blantyre, Malawi. The treatment outcomes were documented from the tuberculosis registers of all patients registered between 1 October 1994 and 30 September 1995. The true treatment outcome for patients who had been registered as defaulters was determined by making personal inquiries at the treatment units and the residences of patients or relatives and, in a few cases, by writing to the appropriate postal address. Interviews were carried out with patients who had defaulted and were still alive and with matched, fully compliant PTB patients who had successfully completed the treatment to determine the factors associated with defaulter status. Of the 1099 patients, 126 (11.5%) had been registered as defaulters, and the true treatment outcome was determined for 101 (80%) of the latter; only 22 were true defaulters, 31 had completed the treatment, 31 had died during the treatment period, and 17 had left the area. A total of 8 of the 22 true defaulters were still alive and were compared with the compliant patients. Two significant characteristics were associated with the defaulters; they were unmarried; and they did not know the correct duration of antituberculosis treatment. Many of the smear-positive tuberculosis patients who had been registered as defaulters in the Blantyre district were found to have different treatment outcomes, without defaulting. The quality of reporting in the health facilities must therefore be improved in order to exclude individuals who are not true defaulters. PMID- 10361756 TI - In vitro sensitivity of Plasmodium falciparum to artesunate in Thailand. AB - Reported are the in vitro susceptibilities of Plasmodium falciparum to artesunate, mefloquine, quinine and chloroquine of 86 isolates and to dihydroartemisinin of 45 isolates collected from areas of high resistance to mefloquine within Thailand near the borders with Myanmar and Cambodia, and from southern Thailand where P. falciparum is generally still sensitive to mefloquine. All the isolates were highly sensitive to artesunate, but the geometric mean IC50S were higher in isolates from the Thai-Myanmar and Thai-Cambodian borders than in those from southern Thailand. The IC50S for mefloquine and artesunate were strongly correlated (Pearson r = 0.605; n = 86; P < 0.00001). As expected, the in vitro sensitivities to dihydroartemisinin and artesunate were similar and strongly correlated (at IC50, Pearson r = 0.695; n = 45; P < 0.00002). The correlation between the activity of mefloquine and artesunate requires further investigation in order to determine the potential for development of cross resistance in nature. Our results suggest that combination with mefloquine is not the ideal way of protecting the usefulness of artemisinin and its derivatives. A search for more suitable partner drugs to these compounds and careful regulation of their use are necessary in the interest of ensuring their long therapeutic life span. PMID- 10361757 TI - Organizing delivery care: what works for safe motherhood? AB - The various means of delivering essential obstetric services are described for settings in which the maternal mortality ratio is relatively low. This review yields four basic models of care, which are best described by organizational characteristics relating to where women give birth and who performs deliveries. In Model 1, deliveries are conducted at home by a community member who has received brief training. In Model 2, delivery takes place at home but is performed by a professional. In Model 3, delivery is performed by a professional in a basic essential obstetric care facility, and in Model 4 all women give birth in a comprehensive essential obstetric care facility with the help of professionals. In each of these models it is assumed that providers do not increase the risk to women, either iatrogenically or through traditional practices. Although there have been some successes with Model 1, there is no evidence that it can provide a maternal mortality ratio under 100 per 100,000 live births. If strong referral mechanisms are in place the introduction of a professional attendant can lead to a marked reduction in the maternal mortality ratio. Countries using Models 2-4, involving the use of professional attendants at delivery, have reduced maternal mortality ratios to 50 or less per 100,000. However, Model 4, although arguably the most advanced, does not necessarily reduce the maternal mortality ratio to less than 100 per 100,000. It appears that not all countries are ready to adopt Model 4, and its affordability by many developing countries is doubtful. There are few data making it possible to determine which configuration with professional attendance is the most cost effective, and what the constraints are with respect to training, skill maintenance, supervision, regulation, acceptability to women, and other criteria. A successful transition to Models 2-4 requires strong links with the community through either traditional providers or popular demand. PMID- 10361758 TI - Nutrition advocacy and national development: the PROFILES programme and its application. AB - Investment in nutritional programmes can contribute to economic growth and is cost-effective in improving child survival and development. In order to communicate this to decision-makers, the PROFILES nutrition advocacy and policy development programme was applied in certain developing countries. Effective advocacy is necessary to generate financial and political support for scaling up from small pilot projects and maintaining successful national programmes. The programme uses scientific knowledge to estimate development indicators such as mortality, morbidity, fertility, school performance and labour productivity from the size and nutritional condition of populations. Changes in nutritional condition are estimated from the costs, coverage and effectiveness of proposed programmes. In Bangladesh this approach helped to gain approval and funding for a major nutrition programme. PROFILES helped to promote the nutrition component of an early childhood development programme in the Philippines, and to make nutrition a top priority in Ghana's new national child survival strategy. The application of PROFILES in these and other countries has been supported by the United States Agency for International Development, the United Nations Children's Fund, the World Bank, the Asian Development Bank, the Micronutrient Initiative and other bodies. PMID- 10361759 TI - Strong bones in later life: luxury or necessity? AB - Osteoporosis is a global problem which will increase in significance as the population of the world both increases and ages. This report looks at how the demographic changes in different countries of the world will be reflected in the incidence and cost of osteoporotic disease. Comparisons are made between the data collected by the European Union's Report on Osteoporosis in the European Community, issued in June 1998, and some of the data available from other parts of the world. The importance of prevention, early detection and appropriate treatment is stressed, as well as the need for national health services to provide reimbursement of the costs of prevention, diagnosis and treatment for high-risk groups. PMID- 10361760 TI - Reducing osteoporosis: prevention during childhood and adolescence. PMID- 10361761 TI - Osteoporosis: a global perspective. PMID- 10361762 TI - Strong bones in later life: luxury or necessity? The view from Brazil:desirable but not yet feasible. PMID- 10361763 TI - Strong bones in later life: luxury or necessity? The view from Tunisia: need for an inclusive approach. PMID- 10361764 TI - Osteoporosis care in Hungary. PMID- 10361765 TI - Vertebral and hip fractures in Japan. PMID- 10361766 TI - Strategies for osteoporosis treatment. PMID- 10361767 TI - Strong bones in later life: luxury or necessity? A patient's perspective. PMID- 10361768 TI - Strong bones in later life: luxury or necessity? The problem of reimbursement. PMID- 10361769 TI - Pioneering community-oriented primary care. PMID- 10361770 TI - The Pholela Health Centre. A progress report. 1952. PMID- 10361771 TI - Simpler screening for cervical cancer. PMID- 10361772 TI - Preventing acute confusion in the elderly. PMID- 10361773 TI - Hip fractures to treble by the year 2030. PMID- 10361774 TI - Antismoking campaigns fail in industrialized countries. PMID- 10361775 TI - Carbamazepine and methylphenidate. PMID- 10361776 TI - Bupropion, periodic limb movement disorder, and ADHD. PMID- 10361777 TI - Looking at the future through orange-colored glasses. PMID- 10361778 TI - Gender identity disorder. PMID- 10361779 TI - French adolescent smoking and perception of quitting. PMID- 10361780 TI - Evolutionary psychology. PMID- 10361781 TI - Psychopathology of childhood social phobia. AB - OBJECTIVE: To describe the clinical syndrome of social phobia in preadolescent children. METHOD: Fifty children with DSM-IV social phobia were assessed with semistructured diagnostic interviews, self-report instruments, parental and teacher ratings, a behavioral assessment, and daily diary recordings. In addition, the behaviors of these children were compared with those of a sample of normal peers. RESULTS: Children with social phobia had a high level of general emotional over-responsiveness, social fear and inhibition, dysphoria, loneliness, and general fearfulness. Sixty percent suffered from a second, concurrent disorder. Socially distressing events occurred quite frequently and were accompanied by maladaptive coping behaviors. In addition, children with social phobia had significantly poorer social skills. There were few differences based on gender or race. CONCLUSIONS: Children with social phobia suffer pervasive and serious functional impairment. In addition, the clinical presentation suggests specific avenues for psychosocial interventions. PMID- 10361782 TI - Suicidal ideation and attempts: a longitudinal investigation of children of depressed and well mothers. AB - OBJECTIVE: This study uses a prospective longitudinal design to examine suicidality (ideation, plans, attempts, and completions) in children and adolescents, to compare suicidality in the offspring of depressed and well mothers, and to identify correlates and predictors of suicidality. METHOD: Two children (n = 192) from each of the families in an ongoing longitudinal study of the offspring of mothers with major depressive disorder (n = 42), with bipolar disorder (n = 26), or without past or current psychiatric diagnosis (n = 30) were studied. Assessment of suicidality, based on diagnostic interviews, was made when the younger of the sibling pairs were approximately 6, 9, and 14 years of age and older siblings were approximately 6, 9, 13, and 18 years of age. RESULTS: Children of depressed mothers were more likely to report suicidal thoughts or behaviors than were children of well mothers (particularly the older sibling cohort). Developmental trajectories of suicidality differed for offspring of mothers with major depressive disorder and bipolar disorder. Links were found between lifetime reports of suicidality and the adolescent's mood problems (e.g., hypomanic behavior), coping strategies, and parental rejection. Also, child's and mother's suicidality were related. CONCLUSIONS: These findings have implications for planning interventions targeted at preventing suicide in youth. PMID- 10361783 TI - Suicide attempts among formerly hospitalized adolescents: a prospective naturalistic study of risk during the first 5 years after discharge. AB - OBJECTIVE: To examine risk for suicide attempts among 180 consecutively referred adolescents during the first 5 years after discharge from an inpatient psychiatry unit. METHOD: In a prospective naturalistic study, adolescents were assessed at psychiatric hospitalization and semiannually thereafter for up to 5 years with semistructured psychiatric diagnostic interviews and self-report questionnaires. RESULTS: Approximately 25% of the adolescents attempted suicide and no adolescents completed suicide within the first 5 years after discharge. The first 6 months to 1 year after discharge represented the period of highest risk. The number of prior attempts was the strongest predictor of posthospitalization attempts. Affective disorders by themselves did not predict later suicide attempts but were related to posthospitalization attempts when accompanied by a history of past suicide attempts. Independent of psychiatric diagnoses, severity of depressive symptoms and trait anxiety also predicted suicide attempts. Similar to the effect with affective disorders, depressive symptoms were most strongly related to posthospitalization suicidality among adolescents with a prior history of suicide attempts. CONCLUSIONS: Particularly among youths with prior suicidal behavior, clinicians should be alert to the above constellation of psychiatric predictors of posthospitalization suicidal behavior. PMID- 10361784 TI - Suicide-bereaved children and adolescents: a controlled longitudinal examination. AB - OBJECTIVE: The current study examined emotional and behavioral sequelae in children who have experienced parental suicide by completing a secondary analysis of data from the Grief Research Study, a longitudinal study of childhood bereavement. METHOD: Twenty-six suicide-bereaved (SB) children, aged 5 to 17 years, were compared with 332 children bereaved from parental death not caused by suicide (NSB) in interviews 1, 6, 13, and 25 months after the death. Children's emotional reactions to the death, psychiatric symptomatology, and psychosocial functioning after the parent's death were determined. RESULTS: Grief emotions were common in both groups. SB children were more likely to experience anxiety, anger, and shame than NSB children. SB children were more likely to have preexisting behavioral problems and more behavioral and anxiety symptoms throughout the first 2 years compared with NSB children. Indices of depression, suicidality, and psychosocial functioning differed minimally between groups. CONCLUSIONS: SB children experience some "common" elements of bereavement. In addition, they demonstrate some lifetime risk factors as well as subsequent pathology that suggests a negative behavioral trajectory. As these cohorts have not yet passed through the age of risk, long-term follow-up is critical. PMID- 10361785 TI - Risk for substance use disorders in youths with child- and adolescent-onset bipolar disorder. AB - OBJECTIVE: Previous work in adults has suggested that early-onset bipolar disorder (BPD) is associated with an elevated risk for substance use disorders (SUD). To this end, the authors assessed the risk for SUD in child- versus adolescent-onset BPD with attention to comorbid psychopathology. METHOD: All youths (aged 13-18 years) with available structured psychiatric interviews were studied systematically. From clinic subjects (N = 333), 86 subjects with DSM-III R BPD were identified. To evaluate the risk for SUD and BPD while attending to developmental issues, the authors stratified the BPD sample into those with child onset BPD (< or = 12 years of age, n = 50) and those with adolescent-onset BPD (13-18 years of age, n = 36). RESULTS: In mid-adolescence, youths with adolescent onset BPD were at significantly increased risk for SUD relative to those with child-onset BPD (39% versus 8%; p = .001). Compared with those with child-onset BPD, those with adolescent-onset BPD had 8.8 times the risk for SUD (95% confidence interval = 2.2-34.7; chi 7(2) = 9.7, p = .002). The presence of conduct disorder or other comorbid psychopathology within BPD did not account for the risk for SUD. CONCLUSIONS: Adolescent-onset BPD is associated with a much higher risk for SUD than child-onset BPD, which was not accounted for by conduct disorder or other comorbid psychopathology. Youths with adolescent-onset BPD should be monitored and educated about SUD risk. The identification and treatment of manic symptomatology may offer therapeutic opportunities to decrease the risk for SUD in these high-risk youths. PMID- 10361786 TI - Relationship of child psychopathology to parental alcoholism and antisocial personality disorder. AB - OBJECTIVE: To evaluate the contributions of familial factors, including parental diagnoses of alcoholism and/or antisocial personality disorder (ASPD), to the risk of developing various child psychiatric diagnoses. METHOD: Four hundred sixty-three children and their biological parents were interviewed with adult and child versions of the Semi-Structured Assessment for the Genetics of Alcoholism. Demographic and psychiatric data were compared across 3 groups of children on the basis of the presence of parental alcoholism and ASPD (no other parental diagnoses were examined). Generalized estimating equations analyses allowed the inclusion of multiple children from each family in the analyses. RESULTS: Among offspring, parental alcoholism was associated with increased risks for attention deficit hyperactivity disorder, conduct disorder (CD), and overanxious disorder. Parental alcoholism plus ASPD was associated with increased risk for oppositional defiant disorder. Dysfunctional parenting style was associated with increased risks for CD, alcohol abuse, and marijuana abuse. Low family socioeconomic status was associated with increased risk for CD. CONCLUSIONS: Parental diagnoses of alcoholism and ASPD were associated with increased risks for a variety of childhood psychiatric disorders, and dysfunctional parenting style was associated with the diagnoses of CD, alcohol abuse, and marijuana abuse. PMID- 10361787 TI - Psychiatric comorbidity among adolescents with substance use disorders: findings from the MECA Study. AB - OBJECTIVE: To investigate the extent to which adolescents in the community with current substance use disorders (SUD) experience co-occurring psychiatric disorders. METHOD: Diagnostic data were obtained from probability samples of 401 children and adolescents, aged 14 to 17 years, and their mothers/caretakers, who participated in the Methods for the Epidemiology of Child and Adolescent Mental Disorders (MECA) Study. RESULTS: The rates of mood and disruptive behavior disorders are much higher among adolescents with current SUD than among adolescents without SUD. Comparison with adult samples suggests that the rates of current comorbidity of SUD with psychiatric disorders are the same among adolescents as adults, and lower for lifetime disruptive disorders/antisocial personality disorder among adolescents than adults. CONCLUSIONS: The high rate of coexisting psychiatric disorders among adolescents with SUD in the community needs to be taken into account in prevention and treatment programs. PMID- 10361788 TI - Long-term stability of Child Behavior Checklist profile types in a child psychiatric clinic population. AB - OBJECTIVE: To study the long-term stability of Child Behavior Checklist (CBCL) profile types, which represent children's overall patterns of single and comorbid scale elevations. METHOD: Profile types were determined for 623 outpatient children at referral and then at mean follow-up 4.8 years later, and their continuity was determined. RESULTS: At baseline 37.5% of the children were classified by a profile type, and 41.9% of these originally classified children continued to be classified at follow-up. The average odds ratio for a child continuing as a specific CBCL profile type from baseline to follow-up was 8.2. When children changed from one specific profile type to another, they usually continued in the same broad externalizing or internalizing category. Children who were not classified by a profile type at baseline generally remained unclassified. CONCLUSIONS: Stability findings for CBCL profile types appeared good and were similar to past longitudinal results for CBCL scales and DSM diagnoses. These profile types may prove an important empirical method for addressing the problem of comorbid clinical pictures. PMID- 10361789 TI - Prevalence and risk factors of behavioral and emotional problems among Chinese children aged 6 through 11 years. AB - OBJECTIVE: To examine the prevalence and risk factors of behavioral and emotional problems in Chinese children. METHOD: A sample of 2,940 children aged 6 through 11 years was randomly drawn from household registers in Shandong Province of China. Parents completed the Child Behavior Checklist (CBCL) and a structured self-rating questionnaire. RESULTS: The mean CBCL Total Problems score was 16.1 (SD = 14.0). There was no significant age effect on the Total Problems score; boys scored significantly higher than girls (17.2 versus 15.0; F = 24.94, p < .01). The overall prevalence rates of behavioral problems were 12.5% for boys and 8.3% for girls (chi 2 = 14.23, p < .01). Logistic regression analysis showed that a number of parental, prenatal, perinatal, and postnatal factors were significantly associated with increased risk of children's behavioral problems. CONCLUSIONS: The prevalence of parent-reported behavioral problems in Chinese children is lower than those found in other countries. Of multiple psychosocial and biological factors associated with children's behavioral problems, separation or divorce of parents is the most significant factor. PMID- 10361790 TI - ADHD in a school sample of Brazilian adolescents: a study of prevalence, comorbid conditions, and impairments. AB - OBJECTIVE: To evaluate the prevalence, comorbid conditions, and impairments of attention-deficit/hyperactivity disorder (ADHD) among young adolescents in Porto Alegre, Brazil. METHOD: 1,013 students aged 12 to 14 years were evaluated at 64 state schools, using a screening instrument based on the 18 DSM-IV ADHD symptoms. All positive screened students (n = 99) and a random subset of negative screened subjects (n = 92) had a psychiatric evaluation carried out within a hospital setting or at home. RESULTS: The prevalence of ADHD was estimated to be 5.8% (95% confidence interval = 3.2-10.6), and the comorbidity with other disruptive behavior disorders was high (47.8%). Youths with ADHD (n = 23) had significantly higher rates of school repetitions, suspensions, and expulsions (p < .01) than controls (n = 168). No association was identified between ADHD and alcohol, marijuana, and inhalant use. CONCLUSION: The results extend to adolescents well documented findings in children, indicating that ADHD is quite prevalent in early adolescence and affected youths are at high risk for impairment and dysfunction in multiple domains. PMID- 10361791 TI - Mental health and social adjustment in young refugee children 3 1/2 years after their arrival in Sweden. AB - OBJECTIVE: To investigate the relative importance of various risk and protective factors for mental health and social adjustment in young refugee children. METHOD: Of 50 Iranian refugee preschool children who were first evaluated 12 months after arriving in Sweden, 39 were reevaluated in a follow-up study 2 1/2 years later. The effect of exposure to organized violence, age, gender, individual vulnerability, parental functioning, and peer relationships on the children's well-being and adjustment was investigated using multiple and logistic regression analyses. RESULTS: Exposure to war and political violence and individual vulnerability before traumatic stress exposure were important risk factors for long-lasting post-traumatic stress symptomatology in children. Mothers' emotional well-being predicted emotional well-being in children, whereas children's social adjustment and self-worth were mainly predicted by the quality of their peer relationships. CONCLUSIONS: The results underline the fact that refugee children's adaptation is the result of a complex process involving several interacting risk and protective factors. For many refugee children, current life circumstances in receiving host countries, such as peer relationships and exposure to bullying, are of equal or greater importance than previous exposure to organized violence. PMID- 10361792 TI - Problems and competencies reported by parents of Vietnamese children in Hanoi. AB - OBJECTIVE: To determine the distribution of behavioral and emotional problems and competencies among a sample of Vietnamese children aged 4 through 18 years living in Hanoi. METHOD: A representative community sample of 1,526 children and adolescents was selected from 2 precincts in Hanoi. Problems and competencies were assessed with the Child Behavior Checklist (CBCL). RESULTS: Vietnamese children had lower mean raw scores than U.S. norms on the CBCL's Total, Externalizing, Internalizing, and Competence scales. Boys were reported to have more externalizing problems and girls more internalizing problems. Girls' levels of internalizing problems increased significantly with age. CONCLUSION: The lower levels of problems and competencies reported in Vietnamese children may represent differences in the prevalence of psychiatric disorders, in parental perceptions of what constitutes deviant behavior, or in parental comfort with reporting psychopathological behaviors. Further research is needed to clarify the relationship between the reported behavioral and emotional problems of Vietnamese children and the presence of psychiatric disorders. From a clinical perspective, the study's results suggest that levels of problems and competencies may vary significantly between different ethnic and cultural groups. Specific clinical cutoffs used to identify children requiring further psychiatric assessment need to be established separately for different ethnic groups. PMID- 10361793 TI - Domains of social communication handicap in autism spectrum disorder. AB - OBJECTIVE: To investigate whether specific "social communication" handicaps could be identified in autism spectrum disorder using the Autism Diagnostic Observation Schedule and to compare the results with those found in a previous factor analysis study using the Autism Diagnostic Interview-Revised. METHOD: All subjects were evaluated with both instruments. J.R. and P.E.T. independently diagnosed autism, Asperger's disorder, or pervasive developmental disorder--not otherwise specified in 51 children. Items from the Autism Diagnostic Observation Schedule that represented social communication behaviors were factor-analyzed. RESULTS: Three factors were identified: joint attention, affective reciprocity, and theory of mind. These are the same social communication domains that were identified in the previous study. CONCLUSIONS: These 3 social communication domains have been discussed in the literature regarding normal development and in previous research on autism spectrum disorders. If these domains are replicated in larger sample sizes, they could be used to monitor the results of pharmacological and psychotherapeutic interventions in autism spectrum disorders. PMID- 10361794 TI - Familial factors influence level of functioning in pervasive developmental disorder. AB - OBJECTIVE: To determine whether siblings with pervasive developmental disorders (PDD) tend to have the same type and number of PDD symptoms or a similar level of functioning. METHOD: The familial correlations for PDD subtype, symptom totals, adaptive behaviors, and nonverbal IQ were calculated for 94 children with PDD from 46 families. RESULTS: On variables measuring PDD symptoms, only impairments in nonverbal communication and verbal/nonverbal status tended to run true within families. There was no familial aggregation of PDD subtype. In contrast, measures of nonverbal IQ and adaptive behaviors in socialization and communication showed a moderate degree of familial resemblance. The degree of familial resemblance did not change if the analysis was restricted only to those families in which both affected children met criteria for autism. CONCLUSION: Insofar as the familial resemblance seen in PDD is due to genetic factors, these data provide some evidence that higher- and lower-functioning PDD children may arise from separate genetic mechanisms. Current gene-mapping studies of PDD may need to take this evidence of genetic heterogeneity into account. PMID- 10361795 TI - Overweight, weight concerns, and bulimic behaviors among girls and boys. AB - OBJECTIVE: To assess the prevalence rates and correlates of overweight, concern with weight, and bulimic behaviors. METHOD: A survey was completed by a population-based sample of 16,114 boys and girls aged 9 to 14 years. RESULTS: Although fewer girls (19%) than boys (26%) were overweight, more girls (25% versus 22%) perceived themselves as overweight (p < .001). The proportion of girls reporting trying to lose weight increased with age (p < .001). The prevalence of binge eating at least monthly increased with age among the girls, but remained stable among the boys. The prevalence of purging was low (< or = 1%) and comparable between genders until age 13. Among the 13- and 14-year-olds, girls were significantly more likely than boys to report using laxatives or vomiting to control weight (p < or = .001). Purging was independently positively associated with stage of pubertal development (girls: odds ratio [OR] = 2.1, 95% confidence interval [CI] 1.6-2.7; boys: OR = 1.5, 95% CI 1.0-2.2) and overweight (girls: OR = 1.9, 95% CI 1.2-3.0; boys: OR = 2.7, 95% CI 1.4-5.1). CONCLUSIONS: Misperception of being overweight and concern with weight were common. Purging was a very rare behavior, but increased with pubertal development. Among the girls, the prevalence increased sharply around the onset of adolescence. PMID- 10361796 TI - Behavior and personality characteristics of children and young adults with Prader Willi syndrome: a controlled study. AB - OBJECTIVE: To analyze (1) which behavior and personality characteristics in Prader-Willi syndrome (PWS) are primarily linked to the syndrome and not to mental retardation or being overweight, (2) how early in life such traits appear, and (3) whether current therapies affect behavior. METHOD: Parents of a group of 44 individuals with PWS and of a comparison group were interviewed and completed questionnaires about their children's behavior and personality. RESULTS: Individuals with PWS had more behavior problems than those in the comparison group. Some behaviors were specific to PWS. Younger PWS cases had fewer behavior problems than older PWS cases. Treated individuals had approximately the same degree of behavior problems as those untreated, even though a few symptoms occurred at lower rates. CONCLUSIONS: PWS is associated with behavior correlates that are not related to weight or IQ. In the first few years of life, children with PWS do not demonstrate the characteristic profile of preoccupation with food, ritualism, irritability, temper tantrums, and skin-picking which is typical of older individuals with PWS. Current therapies (including treatment with growth hormone) do not seem to radically affect the behavioral expression of the disorder, even though some problems tended to abate with treatment. PMID- 10361797 TI - Neuropsychological factors associated with borderline pathology in children. AB - OBJECTIVE: To determine whether children with borderline pathology have a specific pattern of neuropsychological risk factors. METHOD: The subjects were 94 school-age children in day treatment, divided into borderline (n = 41) and nonborderline (n = 53) groups according to results of the Child version of the Diagnostic Interview for Borderlines. All children were assessed with the Child Behavior Checklist, the Schedule for Affective Disorders and Schizophrenia for School-Age Children, and a neuropsychological battery. RESULTS: Children with borderline pathology had abnormal scores on the Wisconsin Card Sorting Test and on the Continuous Performance Test, both of which suggested problems with executive function. Although borderline pathology was highly comorbid with conduct disorder, most results were independent of this comorbidity. CONCLUSIONS: Borderline pathology in children has a unique pattern of neuropsychological risk factors that may reflect a diathesis for this syndrome. PMID- 10361798 TI - Case series: PTSD symptoms in adolescent survivors of "ethnic cleansing." Results from a 1-year follow-up study. AB - OBJECTIVE: The authors describe the psychiatric sequelae of "ethnic cleansing" in adolescent Bosnian refugees, via a 1-year follow-up study. METHOD: Ten Bosnian adolescent refugees from the war in Bosnia-Herzegovina received a baseline assessment within the first year after their resettlement and a follow-up assessment 1 year later. Evaluations included an assessment scale for posttraumatic stress disorder (PTSD) symptom severity. RESULTS: At baseline, 3 subjects met criteria for PTSD. At follow-up, this diagnosis persisted in none of these subjects, though 1 subject met criteria at follow-up only. For the group, mean PTSD severity scores at baseline and at follow-up were 8.9 and 4.0, respectively. At baseline, reexperiencing symptoms were present 43% of the time, avoidance symptoms were present 33% of the time, and hyperarousal symptoms were present 33% of the time; at follow-up, these proportions were 35%, 16%, and 18%, respectively. CONCLUSIONS: Overall, rates of PTSD symptoms diminished during the 1-year follow-up interval, suggesting that they may be transient and not representative of enduring psychopathology. This finding may reflect the relative resiliency of adolescents, as well as a variety of factors that facilitated adaptation in our particular group of adolescent refugees. PMID- 10361799 TI - Cultural influences on symptom presentation in childhood. PMID- 10361800 TI - Research problems and variables. PMID- 10361801 TI - Genetics of childhood disorders: III. Genetics and intelligence. PMID- 10361802 TI - A dynamic model of drug initiation: implications for treatment and drug control. AB - We set up a time-continuous version of the first-order difference equation model of cocaine use introduced by Everingham and Rydell [S.S. Everingham, C.P. Rydell, Modeling the Demand for Cocaine, MR-332-ONDCP/A/DPRC, RAND, Santa Monica, CA, 1994] and extend it by making initiation an endogenous function of prevalence. This function reflects both the epidemic spread of drug use as users 'infect' non users and Musto's [D.F. Musto, The American Disease: Origins of Narcotic Control, Oxford University, New York, 1987] hypothesis that drug epidemics die out when a new generation is deterred from initiating drug use by observing the ill effects manifest among heavy users. Analyzing the model's dynamics suggests that drug prevention can temper drug prevalence and consumption, but that drug treatment's effectiveness depends critically on the stage in the epidemic in which it is employed. Reducing the number of heavy users in the early stages of an epidemic can be counter-productive if it masks the risks of drug use and, thereby, removes a disincentive to initiation. This strong dependence of an intervention's effectiveness on the state of the dynamic system illustrates the pitfalls of applying a static control policy in a dynamic context. PMID- 10361803 TI - Test of mathematical assumptions behind the 'incidence function' estimation process of metapopulations' dynamic parameters. AB - I question Hanski's [I. Hanski, A practical model of metapopulation dynamics, J. Animal Ecol. 63 (1994) 151] assumption that incidence functions are relevant approximations of the equilibrium dynamics of stochastic metapopulation models to estimate models' parameters based on snapshot data. Based on ten different metapopulation models, this assumption is found to be at least partly unjustified when referring to the asymptotic behaviour of the models. This leads me to recommend the use of explicit extinction-colonisation transition probabilities and process data (rather than snapshot data) in the estimation process of metapopulation models. PMID- 10361804 TI - The kinetics of anaerobic metabolism following the initiation of high-intensity exercise. AB - A mathematical study is made of the kinetics of anaerobic metabolism following the initiation of high-intensity exercise. Power and energy relationships are proposed for oxygen-independent glycolysis, phosphocreatine utilisation and the utilisation of endogenous ATP. The power relations consist of two components, one describing the build-up phase, the other the controlled-utilisation phase. The controlled-utilisation phase of oxygen-independent glycolysis and the build-up of aerobic metabolism are shown to be closely inter-related. The theoretical relations display trends consistent with published experimental results. Some property values are derived, but because of the scatter of the experimental results, the values are, in general, to be regarded as tentative. PMID- 10361805 TI - A stochastic model of temporally regulated generation of oligodendrocytes in cell culture. AB - The results of our previous analyses suggest that O-2A progenitor cells become competent for differentiation in vitro after they complete a certain number of critical mitotic cycles. The number of critical cycles varies from clone to clone and should be thought of as a random variable. We propose an approach to the analysis of oligodendrocyte generation in vitro based on a stochastic model allowing for an arbitrary distribution of this random variable with a finite support. When applied to experimental data on clonal growth and differentiation of purified O-2A progenitor cells obtained from optic nerves of 1 and 7 day-old rats, the model provides a good quantitative description not only of the first two moments (mean and variance) of the number of O-2A progenitor cells and oligodendrocytes at different times after the start of experiment, but of the corresponding distributions as well. As our estimates show, there are scarcely any O-2A progenitor cells that divide in vitro more than twice before they acquire the competence for differentiation. Those O-2A cells that have undergone the critical divisions differentiate into an oligodendrocyte in each of the subsequent mitotic cycles with a certain probability. We give estimates of this probability for O-2A cells under different growth conditions. Our analysis suggests that the effect of thyroid hormone is twofold: it reduces the mean duration of the mitotic cycle for progenitor cells, and it increases the probability of their transformation into oligodendrocytes. PMID- 10361806 TI - Some properties of an estimator for the basic reproduction number of the general epidemic model. AB - In this paper some properties of a convenient estimator, derived from a martingale estimating function, for the basic reproduction number of the general epidemic model are given for both finite and large samples. These properties give some guidelines for using this convenient estimator. It is shown that it underestimates the parameter and that the bias tends to zero when the population size and the initial number of infectives are increased simultaneously. The bias cannot be removed for a fixed number of introductory infectives. However, the estimator is asymptotically unbiased, conditional on a major outbreak. A simulation study shows that the central limit theorem applies for moderate population sizes. PMID- 10361807 TI - Growth hormone response to hexarelin, growth hormone-releasing hormone plus pyridostigmine and arginine plus estrogen in prepubertal and early pubertal short children. AB - BACKGROUND: Hexarelin (HEX), a synthetic hexapeptide with a strong GH-stimulating activity, has been suggested as a stimulus for evaluating GH secretion. However, in childhood it has never been compared with other stimuli capable to reduce the effect of the somatostatinergic tone and of the low production of gonadal steroids. METHODS: We evaluated GH response (expressed as the maximum value after stimulus [Cmax] and as area under the curve [AUC], mean +/- SD) to HEX at a dose of 2 micrograms/kg i.v., in comparison with those obtained after GHRH (1 microgram/kg i.v.) + pyridostigmine (PD, 60 mg p.o.) and arginine + ethynylestradiol (E2, 1 mg/day p.o. for 3 days before the test), in 5 subjects with familial short stature (FSS), 11 with constitutional growth delay (CGD), prepubertal (Tanner's stage I) and early pubertal (stage II), and in 8 healthy children age-matched as controls. RESULTS: HEX induced a Cmax of 31.9 +/- 18.4 micrograms/l and an AUC of 1511 +/- 923 micrograms/min x l in stage I, of 36.7 +/ 12.3 micrograms/l and 1938 +/- 903 micrograms/min x l in stage II (ns). GHRH + PD induced a Cmax of 33.8 +/- 14.6 micrograms/l and an AUC of 2072 +/- 1233 micrograms/min x l in stage I, of 29.6 +/- 15.6 micrograms/l and 1901 +/- 1252 micrograms/min x l in stage II (ns). ARG + E2 induced a Cmax of 17.8 +/- 7 micrograms/l and an AUC of 1157 +/- 505 micrograms/min x l in stage I, of 15.6 +/ 11.6 micrograms/l and 649 +/- 452 micrograms/min x l in stage II (ns). The Cmax of HEX was higher than that of ARG + E2 in both stages I and II (p < 0.05); AUC of HEX, was higher than that of ARG + E2 only in stage II (p < 0.01); the Cmax and the AUC of GHRH + PD were higher than those of ARG + E2 both in stage I (p < 0.01 and p < 0.05, respectively) and in stage II (p < 0.05). No difference, neither in the extent of GH response to HEX and GHRH + PD nor in that to stimuli between subjects and controls, was found. HEX has given 32% false positives in stage I and 17% in stage II, GHRH + PD 12% and 15%, while ARG + E2 provided 20% in stage I and 32% in stage II. On the whole, specificity was 76% for HEX and ARG + E2 and 89% for GHRH + PD. CONCLUSIONS: HEX induced greater GH response than that of ARG + E2 but similar to that of GHRH + PD and its specificity was not different to that of ARG + E2 and lower than that of GHRH + PD: then its use does not show a diagnostic advantage in respect to the other two stimuli in peripubertal age. PMID- 10361808 TI - [Leptin and neuropeptide Y serum levls in young obese during weight loss]. AB - BACKGROUND AND AIM: Correlations between serum leptin (LEP) and BMI and the percentage of fat mass (FM), as well as differences between male and female serum levels and their behaviour during weight loss have already been extensively described in adult obesity, whereas few cases have been examined in child and adolescent obesity. There are also few studies of the alterations in NPY in peripheral blood in obese subjects during weight loss. METHODS: This study aimed to evaluate the correlations between LEP and BMI, FM% and NPY in 72 obese subjects, with BMI > 35 (29 males and 43 females) aged between 9.6 and 19.8 years old, during weight loss together with any differences between the sexes. RESULTS: LEP was positively correlated in both sexes with BMI and FM%, whereas no correlation emerged with NPY; LEP levels decreased gradually during weight loss, whereas no changes were observed in NPY except during the first phases of weight loss in males when the decrease was significant. LEP concentrations were significantly higher in females, who also showed a higher FM% with equal BMI. No difference was observed between NPY levels in both sexes. CONCLUSIONS: The authors conclude that: 1) the behaviour of LEP in child-adolescent obesity is broadly comparable to that described in adult obesity; 2) the highest LEP concentrations with equal BMI in females appear to reflect the different body composition of the two sexes given that females have a higher FM%; 3) the control exerted by LEP on hypothalamic NPY cannot be seen in peripheral blood and no differences emerged between the two sexes. PMID- 10361809 TI - [Prevalence of simple sporadic goiter in relation to other non-neoplastic thyroid pathologies in an internal medicine department]. AB - BACKGROUND AND AIM: Thyroid pathologies, in particular goiter, are commonly found in everyday clinical practice in hospitalised patients. METHODS: The authors carried out a prospective type survey lasting 80 months in a mountain community with about 30,000 inhabitants with uniform socio-demographical characteristics. A total of 8,034 subjects (4,131 males and 3,903 females) were included in the study who were hospitalised in an Internal Medicine Department. All patients underwent screening using standard diagnostic criteria. RESULTS: Thyroid pathology was identified and classified into its various forms. There was a net prevalence of goiter which, owing to the unique features identified by the study, was defined as the sporadic form. The study demonstrated a marked prevalence in females, the familial pattern, the homogeneity of distribution throughout the area, the progression of disease with age and its evolution into immersed forms, the prevalence in both sexes in adults, the frequent use of surgery and its inherent problems, the inadequacy of treatment. CONCLUSIONS: The authors affirm that this pathology is regularly underestimated because it is not adequately identified, treated and monitored. They suggest a clinical and methodological approach based on a correct nosological definition, adequate diagnostic and prognostic stratification, an appropriate therapeutic protocol and clinical controls performed over time. PMID- 10361810 TI - [New developments in the treatment of medullary thyroid carcinoma]. AB - Medullary thyroid carcinoma arises from the C cells, which produce a characteristic hormone, calcitonin. At present, surgery is the main treatment modality. Medullary thyroid carcinoma is usually treated with total thyroidectomy and with removal of nodes in the central portion of the neck and upper mediastinum. Cervical nodes dissection may be required for cancers affecting lateral neck nodes. Substitutive therapy with levo-thyroxine is indicated after surgery. External beam radiation therapy is not effective against advanced medullary thyroid carcinoma, while chemotherapy has a marginal activity. Biological therapy induces its anti-tumour activity through the inhibition of tumour cell growth without cytolysis and stimulating the antitumour immune response, in the absence of relevant side effects. On these bases, it can be suggested that chemo-refractory tumours could be still responsive to biological agents. In the last years somatostatin analogues and interferon have been used in the therapy of advanced and symptomatic medullary thyroid carcinoma, demonstrating an efficacious effect on neuroendocrine symptoms and on the production of calcitonin and an improving in quality of life. Even if there are no consistent data on the effects of biological agents on the reduction of tumour mass, the combined use of chemotherapy and biological therapy needs to be experimented in medullary thyroid carcinoma. PMID- 10361811 TI - [Anthropometric and nutritional study in young adults. Evaluation of submandibular skinfold thickness]. AB - OBJECTIVE: To describe the main anthropometric characteristics of a young adult population, to compare some measurements obtained with different formulas, and to compare these results with a reference population. MATERIAL AND METHODS: We have studied 72 healthy female university students aged 19.0 to 20.9 years (mean age 19.63). We have measured weight, height, body mass index (BMI), cervical and arm circumferences, and biceps, triceps, and submandibular skinfolds. We have calculated total arm area, fat arm area and muscular arm area using traditional formulas and others recently published. Means were compared using Student's t test and we also calculated Pearson correlations. RESULTS: Concerning anthropometric measurements, they were similar to those of the reference population. Arm areas were different when calculated with different formulas (p < 0.000). All the anthropometric characteristics measured correlated with each other, except with height. Submandibular skinfold correlated with biceps skinfold (0.467), triceps skinfold (0.513), BMI (0.503), weight (0.476), cervical circumference (0.511) and arm circumference (0.505). Submandibular skinfold also showed a significant correlation with arm measurements, especially fat area (0.519, p < 0.001). CONCLUSIONS: Fat and muscular arm areas were different depending on the different formulas used to be calculated. Submandibular skinfold is easy to obtain and is related with the other indexes of body fat. This measure could be included in nutritional surveys. PMID- 10361812 TI - [Effect of dietary fiber on the nutritional utilization of proteins and minerals]. AB - In the last years, many experimental studies performed in humans and in animals show the relationship between dietary fibre (DF) and lipemia. All agree that fibre supplements are recommendable as a complement during treatments with hypocholesterolemic drug. Nevertheless dietary fibre interacts with the absorption of some nutrients in the diet, and thus on one hand the consumption in adequate amounts may induce the desired effect of lipemia but on the other it may not be advisable from other viewpoints. PMID- 10361813 TI - [Critical analysis of the evolution of commercial preparations for enteral nutrition during 1988-1996]. AB - Enteral nutrition is a form of nutritional support that is continually growing and expanding, and within this area especially the enteral preparations or formulae. The object of the present is to analyze the evolution of the commercially available nutritionally complete enteral preparations between 1988 and 1997, illustrating the variations that have occurred both from the quantitative and form the qualitative points of view. A progressive increase is seen in the absolute number of available enteral formulae, with the increase in both polymeric formulae with dietary fiber, and that of formulae designed for specific diseases being significant (p < 0.05). The standard polymeric formulae and the oligomeric formulae decrease, but not in a significant manner. There is a special discussion or the characteristic and the usefulness of the polymeric formulae with dietary fiber ad the formulae designed especially for specific diseases. The clinical efficacy of most of these special formulae is controversial, with there not being sufficient objective clinical evidence at present that justifies their routine use. PMID- 10361814 TI - [Non-metabolic application of indirect calorimetry]. AB - INTRODUCTION: The metabolic monitoring of the critical patient by means of indirect calorimetry is a technique that is used more and more often in ICU's. OBJECTIVES: To establish the methodological basis for the application of indirect calorimetry in the ventilatory monitoring of the critical patient. METHODS: 20 critical patients with complete support ventilation, who because of their clinical condition required an increased or decreased ventilatory minute volume, are monitored with indirect calorimetry (Deltatrac). The dead space was calculated (Vd/Vt ratio) using the formula; Vd/Vt = 1 - (0.863 x VeCO2/PaCO2 x Vm). The changes in carbon dioxide exhalation (VeCO2) were measured, and so was the dead space after changing the ventilation parameters, and the stabilization time of both parameters was measured also. RESULTS: The average oxygen use (VO2) was 265 +/- 45 ml/min, the average baseline VeCO2 was 219 +/- 38 ml/min, and the mean baseline PaCO2 was 37.8 +/- 8 mm Hg. The mean initial minute volume (Vm) was 10.8 +/- 3.2 l/min; in 10 patients this increased in 74 +/- 20%, and in the rest it decreased by 42 +/- 7%. The time required to reach the new equilibrium was less when the minute volume increased than if this decreased: 45.6 +/- 10 vs. 74.2 +/- 7 min (p < 0.01) for the VeCO2, and 45.4 +/- 7 vs. 76 +/- 6.8 min (p < 0.01) for the PaCO2. Both the PaCO2 and the VeCO2 reached the new equilibrium in similar times. The Vd/Vt prior to the ventilatory change was 0.5 +/- 0.12 and after the change the ratio was 0.49 +/- 0.1; in patients in whom the Vm decreased, the Vd/Vt also decreased (p < 0.01), but in contrast, when the Vm increased, the dead space did too (p < 0.01). CONCLUSIONS: On one hand indirect calorimetry permits monitoring of the metabolic equilibrium, and on the other hand in can monitor the patient's hemodynamics (Fick method) and finally, as has been show by this study, it allows a monitoring of the ventilatory situation of the critical patient with complete supported ventilation. PMID- 10361815 TI - [Evaluation of the body composition by anthropometry and bioelectric impedance in a group of elderly patients recovering from cerebrovascular accidents]. AB - Body composition was assessed by bioelectrical impedance and anthropometry in 25 subjects, 13 men and 12 women aged 68 +/- 9 with approximately 1 year of recovering from stroke. Most of them with a high independence in their diary activities. The main purpose of this study is to know the body composition of elderly patients with this pathology and how affects the two compartments, fat mass and fat free mass when they are measured by two different techniques anthropometry and BIA. Body Mass Index was higher in women than in men and correlation coefficient (r = 0.6) with body fat per cent was similar with both methods: BIA and anthropometry. The body fat per cent values obtained by BIA showed the same trend to be lower for men than for women and in general were higher than the anthropometric values; the high correlation between the body fat per cent by anthropometry and by BIA support this tendency (r = 0.748, p < 0.01). The comparative studies of ours results in elderly subjects recovered from stroke and the literature data in healthy elderly subjects suggests that this pathology do not lean to important changes in body composition. However, further research is necessary to confirm these results. PMID- 10361816 TI - [Evaluation of anorexia in patients with bile duct obstruction]. AB - INTRODUCTION: Obstructive jaundice is often accompanied by protein-caloric malnutrition. The objective of the present study is to analyze the incidence and the degree of alterations in the food ingestion of patients with obstructive jaundice. MATERIAL AND METHODS: In a prospective, cross-sectional study 50 patients with obstructive jaundice (19 benign and 31 malignant) were evaluated. The anorexia was evaluated using Welch's test (subjective evaluation) and by means of quantifying the caloric ingestion. An anthropometric parameter (ideal weigh < 95%) and two biochemical ones (albumin < 3.5 g/dl and pre-albumin < 17 mg/dl) were used to define the degree of malnutrition. RESULTS: 96% of the patients presented alterations in the Welch test and in 72% of the patients the caloric ingestion was below the estimated needs. Overall, the ingestion of food was reduced by 76.3 +/- 30% of the estimated needs (84.7 +/- 28% in the benign cases and 70.9 +/- 32% in the malignant cases). Both the Welch test (r = 0.59; p = 0.01) and the caloric ingestion (r = 0.53; p < 0.001) were inversely correlated with the serum bilirubin. In patients with malnutrition criteria, the caloric ingestion was reduced by 30% against the 12% reduction in the non-malnourished patients (p < 0.05). There was a direct correlation between the two methods used in the assessment of the anorexia (r = 0.71; p < 0.001). CONCLUSIONS: Obstructive jaundice is associated with an important reduction in the caloric ingestion, and this is manifested in both biliary obstructions of a benign origin, and in those of neoplasic origins. PMID- 10361817 TI - [Analysis of artificial nutrition in Catalonia during 1989-19993]. AB - BACKGROUND: To study the evolution of Enteral Nutrition (EN) and Parenteral Nutrition (PN) in Catalonia, Spain, between 1989-1993. METHODS: SAMPLE: all hospitals (60) of level A/B, B, B/C and C level of Catalonia. A mailed questionnaire was used. RESULTS: Between 1989-1993, The Hospital Enteral Nutrition (HEN) increased 72%, 2.15% the PN, 507% the Home Enteral Nutrition (HON) and more then 200% the patients who went out to the hospital with EN. The services that assume the clinical control and the responsibles to prepare the HEN are among a wide narrow. 87% of the hospitals use pharmaceuticals phormulas and 41% of them use only their phormulary, and 75% of the hospitals use feed pumps. The HON was supervised by many different professionals. Important differences were detected between the Higher level and the other levels hospitals in the control, dispensation and administration of the NE. CONCLUSIONS: In 1993 there is not a proper settlement and protocol for the HEN in Catalonia. However, the trends make us a better situation of HEN in the near future. The creation of a multidisciplinary team for nutrition support in those hospitals that are not able to have a specific service is encouraged. PMID- 10361818 TI - [Recurrence. Introduction]. PMID- 10361819 TI - [Genetic factors]. PMID- 10361820 TI - [Posture and behavior factors]. PMID- 10361821 TI - [Therapeutic factors or recurrence of iatrogenic origin]. PMID- 10361822 TI - [Post-therapeutic factors]. PMID- 10361823 TI - [Dental and alveolar dysmorphism]. PMID- 10361824 TI - [Bone dysmorphism]. PMID- 10361825 TI - [The Begg and Tip-Edge technic]. PMID- 10361826 TI - [The straight arch-wire technic]. PMID- 10361827 TI - [The bioprogressive concept and recurrence]. PMID- 10361828 TI - [The Tweed-Merrifield technic]. PMID- 10361829 TI - [Retention: when and how?]. PMID- 10361830 TI - [National survey on the concept of retention]. PMID- 10361831 TI - [The development of concepts and technics]. PMID- 10361832 TI - [Citizen forums: a contribution to the "health democracy"?]. PMID- 10361833 TI - [The potential effects of alcohol on bone mass in menopausal women: review of the literature]. AB - The steady rise in post-menopausal osteoporosis makes it a significant public health problem. The knowledge of risk factors of the illness may allow for developing preventive strategies to implement. Many studies have shown that alcohol consumption is associated with osteoporosis or with fractures, when combined either with falls or poor nutrition. Yet, recent publications have shown that alcohol consumption is associated with a greater bone mass among menopausal women. But there is not much literature relative to this risk factor, and what does exist, is contradictory. This article aims at synthesising the information published on this subject. PMID- 10361834 TI - [Sleep and vigilance in students]. AB - A cross-sectional survey was carried out among students with the higher education system in Paris in November, 1992, during a medical exam carried out during their first year of study. The objective was to better determine the characteristics of sleep, vigilance and prevalence of sleep problems. This survey concerned 3152 students, 52% of them girls. The average age was 20 years +/- 2.38% of students felt they don't sleep sufficiently. Twenty one percent of students maintained that they experience sleep difficulties. Ten percent of students said they are sleepy during the day. Four percent of students take medications for sleeping. 3% of students snore regularly. The practice of a sport, living environment, the duration of daily transportation, remunerated work, and the consumption of stimulants all impact on sleep and/or daytime vigilance. PMID- 10361836 TI - [Screening and management of visual disorders in adolescents: impact on social inequality]. AB - A retrospective cohort study was carried out on a sample of 2625 Belgian's children, examined at 12 and 15 year, to analyze the impact of school visual screening on social inequalities for visual acuity. Although uncorrected visual acuity decreased with increase in social class, visual acuity tested with the child's usual correction increased with social class. This social inequalities became more pronounced 2 years after the school visual screening. The lack of accessibility at some levels of health care cancels the reduction of inequalities expected from the school screening. PMID- 10361835 TI - [How are palliative care needs estimated in short-stay establishments? Apropos of an experience in Cote d'Or]. AB - The evaluation of needs and means concerning the care of patients in palliative treatment is among the problems seen as priority by the High Committee for Public Health. We have thus tried to characterise, in a specific health sector, the patients receiving palliative care in short-stay establishments in order to evaluate their care needs. We carried out an exhaustive descriptive survey among all public and private short-stay establishments in the Cote d'Or region. Of the 2116 patients in the hospital on the day of the survey, roughly 30% were considered as eligible for palliative care (patients suffering from serious, chronic and progressive illnesses). The average age of these patients is 63.9 years (standard deviation 19.7). They mainly suffer from tumours (50%), circulatory pathologies (15%), mental illness (7%), or neurological illnesses (6%). Among the patients that may need palliative care, 38% say they feel pain despite treatment with pain-killers among 25.3% of them. The personnel providing care is insufficiently trained in palliative care or in pain, as only a maximum of 18% of nurses and 5% of doctors in short-stay establishments have been trained in these areas. PMID- 10361837 TI - [The private hospital system before and after the 24 April 1996 regulations]. AB - The hospital system has seen itself transformed by the regulations of April 24, 1996. The appearance of new structures and a new logic modified the links between actors: recourse to the contract became an instrument of regulation, the regionalization through the creation of ARH (regional hospitalisation agencies) defined in a new manner the responsibilities in matters of hospital management, the implementation of the ANAES is accompanied by the implementation of an accreditation procedure and pursues the missions of evaluations that, until now, were ensured by the ANDEM. PMID- 10361838 TI - [A feasibility study of an Internet site on health conferences and regional health programs]. AB - With the regulations of April 1996, national and regional health conferences, as well as regional health programmes were created and became mandatory within the health system. In order to promote this multi-disciplinary approach to public health, the Director General for Health and the National School of Public Health wish to distribute, via Internet, all documents developed during these meetings. This data base will be a tool for the groups of regional health programmes and a source of information for both actors in health and consumers. It will be developed in two steps, with the development of a model throughout four regions and then with an overview of the country. This article describes the project approach, the difficulties experienced and the prospects for this data base. PMID- 10361839 TI - [Diagnosis of health in a population and humanitarian actions: a practical guide]. AB - The identification of priority public health problems is the objective of a population health diagnosis. We propose four stages for its development. 1. A description of the situation gives information about the socio-demographic characteristics, sociological, economic, and health indicators, and cultural and economic determinants that influence the health of the population in question. 2. The analysis of consultations among the population and professionals gives indications as to the existence, or lack, of significant collective health problems (diagnosis of perception). 3. From the available statistics, the study of epidemiological data may corroborate the previously obtained results (objectified diagnosis). 4. The outcome of these two approaches may lead to a combination of the diagnosis of perception and the objectified diagnosis and permits, with the help of priority criteria, to determine the priority problems. The approach and the tool presented here were tested among thirty teams responsible for humanitarian programmes and are the result of three years of collaboration with Medecins du Monde. The objective of this approach is to provide the base for conceptual reflection on humanitarian interventions as community action and to propose some elements of methods. PMID- 10361841 TI - Application of two neural network paradigms to the study of voluntary employee turnover. AB - Two neural network paradigms--multilayer perceptron and learning vector quantization--were used to study voluntary employee turnover with a sample of 577 hospital employees. The objectives of the study were twofold. The 1st was to assess whether neural computing techniques offered greater predictive accuracy than did conventional turnover methodologies. The 2nd was to explore whether computer models of turnover based on neural network technologies offered new insights into turnover processes. When compared with logistic regression analysis, both neural network paradigms provided considerably more accurate predictions of turnover behavior, particularly with respect to the correct classification of leavers. In addition, these neural network paradigms captured nonlinear relationships that are relevant for theory development. Results are discussed in terms of their implications for future research. PMID- 10361840 TI - [Contempt and hatred: sociologic indications for an approach to current juvenile violence]. AB - Violence is present in all forms of society that impose limits that must not be surpassed (murder, incest ...) in order for social life to be possible. But these limits are always broken. Youth are, in this respect, in a difficult situation, especially those from marginalized areas. Victims of contempt and exclusion, they react through violent acts; to violence experienced because of excessive constraints in school, they respond with violence. This juvenile violence calls for macro-social responses that return credibility to the institutional system often corrupt and responsible for hidden social violence. It also calls for micro social actions, especially within the community. These actions may allow for awareness of violence generating phenomena, the discovery of obstacles to exercising rights, and the make-up of conflicts and their resolution in negotiation procedures. PMID- 10361842 TI - Evaluating sex discrimination claims: the mediating role of attributions. AB - The role of attributions in judgments of sex discrimination was examined in 2 laboratory experiments. In Study 1, participants read 1 of 12 brief scenarios in which limited information about the strength of evidence against a fictitious corporation and occupational gender stereotype were manipulated. Results suggested that attributions mediated the relationships between participants' gender, strength of evidence, and discrimination judgments. In Study 2, participants were provided with 1 of 3 detailed, typewritten summaries of evidence presented in a sex discrimination trial. Results indicated that jurors' gender was again significantly related to attributions and to sex discrimination judgments even in the face of substantial objective information related to the case. The variance in observers' judgments associated with gender, however, appeared to be greatest when information about the organization's guilt or innocence was equivocal. PMID- 10361843 TI - Ultrasonographic evaluation of the common bile duct in biliary acute pancreatitis patients: comparison with endoscopic retrograde cholangiopancreatography. AB - We compared the morphologic findings of the common bile duct by ultrasonography and endoscopic retrograde cholangiopancreatography in patients with biliary acute pancreatitis. Forty-five patients were studied. The diagnosis of acute pancreatitis was based on the presence of characteristic abdominal pain associated with an elevation of serum amylase and lipase concentrations. All patients underwent ultrasonography and subsequently urgent endoscopic retrograde cholangiopancreatography and eventually endoscopic sphincterotomy. Ultrasonography showed gallstones in 33 patients and sludge of the gallbladder in seven patients. In the common bile duct, lithiasis was found in two patients and sludge in 25. Endoscopic retrograde cholangiopancreatography showed choledocolithiasis in eight patients and sludge of the common bile duct in 32. In 27 cases (60%) concordance occurred between ultrasonographic and endoscopic retrograde cholangiopancreatographic detection of lithiasis or sludge of the common bile duct. The average diameter of the common bile duct determined by sonography was significantly smaller (P < 0.001) than that obtained by endoscopic retrograde cholangiopancreatography. The evaluation of this parameter indicated that a good correlation existed between the values obtained with the two techniques (r(s) = 0.765, P < 0.001). Both ultrasonography and endoscopic retrograde cholangiopancreatography can provide reliable measurements of the common bile duct diameter. Ultrasonography is the technique of choice in the initial investigation of patients with biliary acute pancreatitis. PMID- 10361844 TI - Ultrasonographic arterial portography with second harmonic imaging: evaluation of hepatic parenchymal enhancement with portal venous flow. AB - Ultrasonographic arterial portography was evaluated with second harmonic and conventional gray scale imaging after the administration of 0.001 to 0.1 ml/kg of FS069 (Optison) in 10 dogs (four dogs with ligation of the portal vein branch) and two woodchucks with hepatocellular carcinomas. Harmonic imaging was required to obtain good liver parenchymal enhancement for ultrasonographic arterial portography to be useful. The tumors were visible as regions of greater enhancement after intravenous injection and as hypoechoic regions after superior mesenteric artery injection. The segments with portal vein ligation were not detected after intravenous injection but were clearly seen after superior mesenteric artery injection. Doppler signal measurement verified a significant difference between the portal vein and hepatic vein after superior mesenteric artery injection and in the femoral artery after intravenous versus superior mesenteric artery injection, demonstrating that minimal levels of FS069 pass through the liver. PMID- 10361845 TI - Ring-down artifacts posterior to the right hemidiaphragm on abdominal sonography: sign of pulmonary parenchymal abnormalities. AB - The aim of our study was to verify whether ring-down artifacts posterior to the right hemidiaphragm on abdominal sonography reflected pulmonary parenchymal abnormalities. Forty patients (group 1) with abdominal diseases and 32 patients (group 2) with proved various pulmonary abnormalities involving the right lung base underwent abdominal sonography with 2-4 MHz transducers. In these two groups, the presence and number of ring-down artifacts were assessed and correlated with peridiaphragmatic lung findings on chest radiographs or computed tomographic scans. In 21 patients (group 3) with multiple (more than five) or numerous (10 or more) ring-down artifacts, chest radiographs were reviewed to see if any peridiaphragmatic pulmonary abnormalities were present. In group 1, one or several (less than five) ring-down artifacts were shown in 27 of 40 (68%) patients. In these patients, computed tomography showed insignificant focal intra and interlobular septal thickening in the peridiaphragmatic right lung. In group 2, 31 of 32 (97%) patients showed multiple or numerous ring-down artifacts. In group 3, chest radiographs showed various pulmonary abnormalities in 20 of 21 (95%) patients, including emphysema, idiopathic interstitial pneumonia, bronchopneumonia, and interstitial edema. Although nonspecific, ring-down artifacts posterior to the right diaphragm on abdominal sonography may be used to predict pulmonary abnormalities when encountered on abdominal sonography in patients without specific pulmonary symptoms. PMID- 10361846 TI - Ultrasonographic diagnosis and color flow Doppler sonography of internal jugular venous ectasia in children. AB - We investigated the diagnostic utility of ultrasonography in the diagnosis of internal jugular venous ectasia. Eight children (six boys, two girls) were recruited into this prospective study. Sonography of internal jugular venous ectasia in these patients revealed fusiform dilation of the internal jugular vein, and the possibility of thrombus and external compression could be ruled out. Marked variation in size of ectatic jugular veins during respiration was demonstrated under real-time sonography. The mean anteroposterior diameter of these dilated internal jugular veins was 0.79+/-0.18 mm (mean+/-standard deviation), which increased to 1.58+/-0.27 mm with Valsalva maneuver. Our study showed that the anteroposterior diameters of the internal jugular veins in cases of ectasia were greater than those of contralateral jugular veins in same patients as well as those in normal children, and they showed greater increase after Valsalva maneuver. Under color Doppler flow studies, turbulent vascular flows were demonstrated in these patients with jugular venous ectasia. No progression of venous ectasia was found in any of our patients during a 6 month follow-up period. We conclude that internal jugular venous ectasia in children is a benign condition, which usually does not require surgical intervention. Ultrasonography is a good diagnostic modality for the diagnosis of internal jugular venous ectasia. Color Doppler ultrasonography demonstrate the turbulent flow in jugular venous ectasia. PMID- 10361847 TI - Portal venous system aneurysms: report of five cases. AB - Until recently aneurysms in the portal venous system were considered to be very rare lesions. This opinion has largely been changed by the increasing number of cases reported in recent years. In this paper we report the cases of five patients with portal venous system aneurysms, including one with splenic vein aneurysm. One patient had associated portal hypertension. The reexamination of two patients 2 years later showed no change in the aneurysms. The sonographic features and related literature are reviewed. In the light of this series and the information in the literature, we recommend that portal venous system aneurysms should no longer be considered exceptionally rare entities. PMID- 10361848 TI - Umbilical venous pulsation indicating tight cord entanglement in monoamniotic twin pregnancy. PMID- 10361849 TI - Prenatal diagnosis of a coronary fistula in a fetus with pulmonary atresia with intact ventricular septum and trisomy 18. PMID- 10361850 TI - Sonographic diagnosis of recurrent ulnar nerve compression by ganglion cysts. PMID- 10361851 TI - Congenital splenorenal venous shunt detected by prenatal ultrasonography. PMID- 10361852 TI - Vesicouterine fistula after cesarean section: ultrasonographic findings in two cases. PMID- 10361853 TI - Expression of the MRP2 gene-encoded conjugate export pump in human kidney proximal tubules and in renal cell carcinoma. AB - Human kidney proximal tubule epithelia express the ATP-dependent export pump for anionic conjugates encoded by the MRP2 (cMRP/cMOAT) gene (symbol ABCC2). MRP2, the apical isoform of the multidrug resistance protein, is an integral membrane glycoprotein with a molecular mass of approximately 190 kD that was originally cloned from liver and localized to the canalicular (apical) membrane domain of hepatocytes. In this study, MRP2 was detected in human kidney cortex by reverse transcription-PCR followed by sequencing of a 826-bp cDNA fragment and by immunoblotting using two different antibodies. Human MRP2 was localized to the apical brush-border membrane domain of proximal tubules by double and triple immunofluorescence microscopy including laser scanning microscopy. The expression of MRP2 in renal cell carcinoma was studied by reverse transcription-PCR and immunoblotting in samples from patients undergoing tumor-nephrectomy without prior chemotherapy. Clear-cell carcinomas, originating from the proximal tubule epithelium, expressed MRP2 in 95% (18 of 19) of cases. Immunofluorescence microscopy of MRP2 in clear-cell carcinoma showed a lack of a distinct apical-to basolateral tumor cell polarity and an additional localization of MRP2 on intracellular membranes. MRP2, the first cloned ATP-dependent export pump for anionic conjugates detected in human kidney, may be involved in renal excretion of various anionic endogenous substances, xenobiotics, and cytotoxic drugs. This conjugate-transporting ATPase encoded by the MRP2 gene has a similar substrate specificity as the multidrug resistance protein MRP1, and may contribute to the multidrug resistance of renal clear-cell carcinomas. PMID- 10361854 TI - Neutral lipid from proteinuric rat urine is a novel inhibitor of the red blood cell calcium pump. AB - Proteinuria may be associated with hypertension and progression of renal insufficiency, which in turn may accompany abnormalities in cell calcium homeostasis. Therefore, urine from rats made proteinuric by puromycin aminoglycoside administration was analyzed, in a search for factors affecting cellular calcium transport. Proteinuric urine was fractionated by thin-layer chromatography and HPLC, and the effects of the fractions on the plasma membrane calcium pump in human red blood cells were assessed. Proteinuric urine contained a powerful specific inhibitor of the calcium pump that had little or no effect on the Na+/K+- or Mg2+-ATPases. The inhibitor was characterized as a neutral lipid, migrating as a single band, that inhibited 45Ca2+ efflux. To confirm the presence of an inhibitor in other proteinuric states, the urine from two patients with proteinuria was examined and subjected to chromatography as in the rat studies. These thin-layer chromatographic fractions contained a very strong inhibitor of the red blood cell calcium pump, suggesting that this substance may have relevance for the pathogenesis of proteinuric renal disease in human patients. Rat proximal tubule cells in tissue culture, when challenged with lipid-replete albumin, secreted an inhibitor of the calcium pump that migrated in the same chromatographic band as the urine factor. Therefore, the processing of fatty acids borne by albumin into endocytosing proximal tubular epithelium results in the synthesis and release of a previously unknown lipid modulator of the calcium pump, an effect that may predispose kidney tissue toward elevations in cytosolic calcium levels in target cells. PMID- 10361855 TI - Neuropeptide Y and ATP interact to control renovascular resistance in the rat. AB - Neuropeptide Y (NPY) and ATP are cotransmitters of norepinephrine (NE). Modulation of ATP-mediated purinergic neurotransmission by NPY was investigated in rat perfused kidney. Beta,gamma-Methylene-L-ATP (beta,gamma-mATP; 1.0 to 1.5 microM, n = 8), NE (0.1 microM, n = 8), and NPY (0.1 microM, n = 14) increased perfusion pressure by maximally 12 +/- 1, 17 +/- 2, and 9 +/- 1 mmHg, respectively. In the presence of NPY, responses to ATP and NE were dramatically enhanced. Renal nerve stimulation in the presence of the alpha-adrenoceptor antagonist phentolamine (1 microM) induced pressor responses of 54 +/- 5 mmHg (n = 6). Alpha-blockade-resistant responses were abolished by the P2-purinoceptor blocker suramin (300 microM) and thus mediated by ATP. Purinergic responses were also reduced significantly (50%) by the NPY-Y1 receptor blocker BIBP 3226 (1 microM). NPY (0.1 microM) potentiated purinergic pressor responses and enhanced ATP release from 0.7 +/- 0.2 to 4.1 +/- 0.9 pmol (n = 4) associated with a significant increase of soluble ATPase activity. All NPY effects were prevented by BIBP 3226. Pressor responses to renal nerve stimulation delivered at short time intervals, mimicking enhanced sympathetic drive to the kidney, were not constant but showed a progressive rise, which was prevented by BIBP 3226. In this study, it is suggested that purinergic vasoconstriction in rat kidney depends on concomitantly released NPY. NPY by itself is only a weak vasoconstrictor but acts as a modulator of renal vascular resistance by enhancing the effects of its sympathetic cotransmitters, especially during sympathetic overactivity. PMID- 10361856 TI - Evidence of a role for Ki-Ras in the stimulated proliferation of renal fibroblasts. AB - Progressive renal fibrosis is driven by a range of cytokines that act via membrane receptors and intracellular signaling cascades to evoke gene transcription events and related responses. The Ras family of GTPases has been implicated in many of these signaling cascades in model systems such as 3T3 fibroblasts. However, the roleof the specific Ras isoforms Ki, Ha, and N in the stimulation of renal fibroblasts has not been defined. In this study, Ras has been inhibited in primate renal fibroblasts (vero cells) using specific phosphorothioate oligodeoxynucleotides (oligos) targeting the three isoforms. Lipofectin transfection with 200 to 400 nM Ki-Ras oligo inhibited the epidermal growth factor- and fibroblast growth factor-stimulated proliferation of vero cells by 25 to 35% with a lesser effect on serum-stimulated growth. Oligos against Ha-Ras and N-Ras were inactive with respect to control oligo. Total cellular Ras protein (estimated by Western blotting) was reduced by 60 to 90% 24 h after transfection with Ki-Ras oligo. N-Ras, Ha-Ras, and control oligos were inactive. Total Ras synthesis over 4 h measured using [35S]-cys/met pulse chase was reduced by approximately 70% by Ki-Ras oligo and not altered by other oligos. The fractional prenylation of Ras was quantified from the discrete bands on polyacrylamide gel electrophoresis and was increased by the Ki-Ras oligo alone. These data demonstrate that these renal fibroblasts predominantly express the Ki isoform of Ras and that this GTPase plays a role in the stimulated proliferation of these cells. Ras GTPases may be a target for the inhibition of processes leading to renal fibrosis. PMID- 10361857 TI - Effect of high glucose on mesangial cell protein kinase C-delta and -epsilon is polyol pathway-dependent. AB - In diabetes mellitus, enhanced activity of mesangial cell protein kinase C (PKC) may contribute to nephropathy. The purpose of this study was to determine whether high glucose alters mesangial cell diacylglycerol-sensitive PKC-alpha, -beta2, delta, and -epsilon content, cellular distribution, and activity through polyol pathway activation. Primary cultured rat mesangial cells (passage 10) were growth arrested in 0.5% fetal bovine serum and cultured in 5.6 mM glucose (NG) or 30 mM glucose (HG) for 48 h, with or without the aldose reductase inhibitor tolrestat or ARI-509. PKC isoform content in total cell lysates, or cytosol, membrane (Triton X-soluble), and particulate (sodium dodecyl sulfate-soluble) fractions was analyzed by immunoblotting, and band density in HG was expressed as a percentage of corresponding NG values. In HG at 48 h, increased total PKC-alpha (222 +/- 17% of NG, P < 0.001), -beta2 (209 +/- 12%, P < 0.001), and -epsilon (195 +/- 19%, P < 0.001) were observed. L-Glucose had no effect on total PKC isoform content. HG caused increased membrane- and particulate-associated PKC alpha (257 +/- 87 and 327 +/- 66%, respectively, P < 0.05), membrane-associated PKC-delta (143 +/- 10%, P < 0.05), and membrane-associated PKC-epsilon (186 +/- 11%, P < 0.001), with no change in cytosol contents. The HG effects were not mimicked by L-glucose. In NG or HG, PKC-beta2 was not detected in the cytosol fraction, and membrane and particulate association were unchanged with phorbol ester stimulation. Confocal immunofluorescence imaging revealed that in HG, PKC alpha, -delta, and -epsilon translocate to the nucleus and plasma membrane. Total PKC activity measured by in situ 32P-phosphorylation of the epidermal growth factor receptor substrate increased from 18 +/- 1 pmol/min per mg cell protein in NG to 33 +/- 3 pmol/min per mg cell protein in HG (P < 0.002 versus NG). In NG, tolrestat and ARI-509 exposure caused increased PKC activity, enhanced accumulation of total PKC-alpha and -beta2, with no change in total or fractional recovery of PKC-delta or -epsilon. In HG, tolrestat and ARI-509 prevented the increase in total PKC-epsilon and membrane-associated PKC-delta and -epsilon. It is concluded that within 48 h of HG, enhanced mesangial cell PKC activity is associated with accumulation and cellular redistribution of diacylglycerol sensitive PKC isoforms, and that increased PKC-epsilon content and membrane associated PKC-delta and -epsilon are dependent on polyol pathway activation. PMID- 10361858 TI - Induction of monocyte chemoattractant protein-1 by albumin is mediated by nuclear factor kappaB in proximal tubule cells. AB - The transcription and translation of monocyte chemoattractant protein-1 (MCP-1), a CC chemokine, are increased in proximal tubule epithelial cells (PTC) stimulated with pathophysiologically relevant concentrations of albumin. The purpose of this study was to investigate whether nuclear factor kappaB (NFkappaB)/Rel proteins play a role in albumin-induced MCP-1 transcription. Confluent monolayers of rat PTC in primary culture were stimulated with delipidated bovine serum albumin. NFkappaB, the NFkappaB inhibitory protein (IkappaB), and MCP-1 transcription were assessed using electrophoretic mobility shift assays, Western immunoblotting, semiquantitative reverse transcription-PCR, and ribonuclease protection assays. Activation of NFkappaB by delipidated bovine serum albumin (15 mg/ml) was detectable within 2 h, maximal after 8 h, and maintained for at least 16 h of continuous exposure. Supershift analysis showed that the activated proteins were composed of p50/p50, p50/p65, and p50/c-Rel dimers. dimers. Cytoplasmic IkappaBalpha levels were decreased 30 min after stimulation and returned to unstimulated levels by 4 to 8 h. IkappaBbeta levels were decreased at 2 h and there was no recovery until 8 h. Inhibition of NFkappaB with pharmacologic agents (N-tosyl-phenylalanine chloromethyl ketone and dexamethasone) and an antisense oligonucleotide to the rat p65 subunit of NFkappaB significantly reduced MCP-1 transcription. The 3.6-kb 5' flanking region of the rat MCP-1 gene was cloned and sequenced, and two putative kappaB binding sites were identified within the enhancer region. Therefore, albumin increased NFkappaB and reduced IkappaB levels in PTC, and MCP-1 expression was dependent on NFkappaB activation. It is concluded that the activation of NFkappaB/Rel proteins modulates chemokine production in PTC in response to albumin and is likely to have an important role in the mediation of tubulointerstitial injury in proteinuric renal disease. PMID- 10361859 TI - Binding of the renal epithelial cell line LLC-PK1 to laminin is regulated by protein kinase C. AB - The alpha6beta1 integrin heterodimer has been implicated in the mediation of renal epithelial cell binding to laminin, and it has been suggested that this binding is important for renal morphogenesis and development. Studies of nonrenal cells have suggested that the functional activity of alpha6beta1 integrin is regulated by protein kinase C (PKC) activity. In this study, the binding of a renal epithelial cell line, LLC-PK1, to laminin was characterized and the role of PKC activity in the modulation of binding was investigated. LLC-PK1 cells bound to laminin-coated surfaces in a time- and laminin concentration-dependent manner. Binding was strongly inhibited by anti-beta1 integrin antibodies and by anti alpha6 integrin antibodies. Antibodies against alpha2 integrin and a3 integrin had little inhibitory effect. Cells bound to both whole laminin and laminin fragment E8, i.e., the fragment to which the alpha6beta1 integrin heterodimer binds. Exposure of cells to PKC activators for as little as 2 h enhanced cell binding to laminin approximately twofold, in a protein synthesis-dependent manner. PKC inhibitors antagonized this effect. PKC-stimulated binding was also inhibited by anti-beta1 integrin and anti-alpha6 integrin antibodies. PKC activation did not alter expression of beta1 integrin subunits at the cell surface after short time periods (2 to 4 h), but expression was increased after longer time periods (24 h). These results indicate that the renal epithelial cell line LLC-PK1 binds to laminin via the alpha6betal integrin heterodimer and binding is enhanced by PKC activation. The PKC-mediated enhancement of binding requires protein synthesis and is mediated in part by activation of surface alpha6beta1 integrin. PMID- 10361860 TI - A large subset of neutrophils expressing membrane proteinase 3 is a risk factor for vasculitis and rheumatoid arthritis. AB - It has been shown previously that proteinase 3 (PR3), a neutrophil intracellular protease that is the main antigen of antineutrophil cytoplasm (ANCA) autoantibodies, is present on the plasma membrane of a subset of freshly isolated neutrophils. This study shows that the size of this subset of membrane PR3 positive (mPR3+) neutrophils is a stable feature of a given individual, most likely genetically controlled. It ranges from 0 to 100% of neutrophils and allows us to define a new polymorphism in the healthy population, with three discrete phenotypes corresponding respectively to less than 20% mPR3 + neutrophils (mPR3low) or to a mean percentage of 47% (mPR3intermediate) and 71.5% (mPR3high) mPR3+ neutrophils. The frequency of the mPR3high phenotype was significantly increased in patients with ANCA-associated vasculitis (85% versus 55% in healthy subjects). The percentage of mPR3+ neutrophils was not affected by disease activity, relapses, or therapy, and did not reflect in vivo cell activation. In addition, mPR3+ phenotypes were normally distributed in cystic fibrosis patients, indicating that infection and/or inflammation per se do not lead to a high percentage of mPR3+ neutrophils. The frequency of the mPR3high phenotype was not related to anti-PR3 autoimmunization, since it was increased in vasculitic patients regardless of the ANCA specificity (anti-PR3, anti-myeloperoxidase, or unknown). Interestingly, the frequency of the mPR3high phenotype was also increased in patients with rheumatoid arthritis. It was normal in type I diabetes, a T cell-dependent autoimmune disease. It is proposed here that a high proportion of membrane PR3-positive neutrophils could favor the occurrence or the progression of chronic inflammatory diseases. PMID- 10361861 TI - Unrecognized pattern of von Willebrand factor abnormalities in hemolytic uremic syndrome and thrombotic thrombocytopenic purpura. AB - Heterogeneous abnormalities in multimeric structure and fragmentation of endothelial-derived von Willebrand factor (vWF) have been reported in hemolytic uremic syndrome (HUS) and thrombotic thrombocytopenic purpura (TTP). This study was conducted to establish whether different patterns of vWF abnormalities were associated with different clinical syndromes. Plasmatic levels of vWF antigen (vWF:Ag), vWF release from endothelial cells (EC) exposed to patient sera, and vWF multimeric pattern were studied during episodes and again in remission in three groups of patients with severe forms of HUS and TTP paradigmatic of the most common clinical patterns of disease presentation: (1) plasma-responsive; (2) plasma-resistant; and (3) frequently relapsing. Plasma vWF:Ag and serum-induced vWF release from EC were increased in the acute phase of either plasma-responsive and plasma-resistant HUS and TTP, but normalized at remission only in plasma responsive cases. Both indices were persistently normal in the relapsing forms. Enhanced vWF fragmentation as defined by disappearance of high molecular weight and increase in low molecular weight forms was a consistent finding of the acute phases, and always normalized in remission in all three groups. Unusually large vWF multimers were found exclusively in plasma of relapsing forms of HUS and TTP both during and between relapses. Enhanced levels of vWF:Ag and serum capability to induce vWF release in vitro are markers of disease activity and may reflect systemic endothelial injury and consequent activation. Their presence discriminates acute single-episode cases from relapsing forms and, when failing to normalize with plasma therapy, predicts plasma resistance. Enhanced low molecular weight multimers that closely paralleled disease activity suggest a permissive role of fragmented vWF in the formation of microvascular thrombi. Finally, finding of unusually large multimers exclusively in relapsing forms of HUS and TTP even between relapses, when no other clinical signs of disease activity could be detected, suggests that they cannot be the only factor in microvascular thrombosis. PMID- 10361862 TI - The terminal sequence of complement plays an essential role in antibody-mediated renal cell apoptosis. AB - Mesangial cell (MC) injury is a characteristic feature in the early phase of Thy.1 nephritis. The present study investigates the contribution of complement to MC apoptosis in this experimental model of kidney disease in rats. Thy.1 nephritis was induced by injection of mouse anti-Thy.1 monoclonal antibody (ER4G). To assess the contribution of the terminal sequence of complement on apoptosis, the studies were performed in complement-sufficient PVG/c (PVG/c+) rats and in rats deficient in complement C6 (PVG/c-). Apoptosis was monitored by assessment of the number of condensed nuclei in kidney sections stained with periodic acid-Schiff (PAS) and by the terminal deoxynucleotidyl transferase mediated nick end labeling (TUNEL) method and expressed as number of apoptotic cells per 50 glomerular cross sections. In the PAS method, 1 h after intravenous injection of ER4G, PVG/c+ rats exhibited 160.9 +/- 49.5 apoptotic cells, whereas PVG/c- rats had only 3.2 +/- 1.4 apoptotic cells. Control rats exhibited 0.9 +/- 0.6 apoptotic cells. These findings were confirmed with the TUNEL method. In PVG/c- rats, a maximum number of 8.8 +/- 3.1 TUNEL-positive (TUNEL+) cells was found at 6 h followed by a decline thereafter. In PVG/c+ rats, apoptosis was associated with deposition of C6 and C5b-9. Restoration of the complement system of PVG/c- rats with purified human C6 resulted in an increase of apoptosis at 1 h after injection of ER4G from minimal numbers to 239.9 +/- 52.4 TUNEL+ cells. These studies appear to indicate for the first time that the terminal sequence of complement is involved in induction of apoptosis. PMID- 10361863 TI - Expansion of cortical interstitium is limited by converting enzyme inhibition in type 2 diabetic patients with glomerulosclerosis. The Diabiopsies Group. AB - Renal interstitial expansion is now considered a useful marker of progression of several nephropathies. This study describes a multicenter, prospective, double blind, placebo-controlled, randomized trial of the effects of Perindopril (4 mg/d) on kidney structure and function over 2 yr in 26 type 2 diabetic patients with proteinuria ranging from 70 to 4210 mg/d and relatively preserved GFR (creatinine clearance >60 ml/min). All patients underwent baseline renal biopsy, but four (15%) were not randomized because of the presence of nondiabetic nephropathy. The remaining 22 were randomized ( 11 to Perindopril [PE], 11 to placebo [PO]), and 19 (9 PE, 10 PO) underwent follow-up biopsy at 2 yr. BP was controlled equally in both groups throughout. Proteinuria increased in PO patients (+1562 mg/d) but declined in PE patients (-156 mg/d) (P < 0.05). Morphometric analysis was performed by light microscopy using a Biocom computer. Over the 2 yr, mean cortical interstitial fractional volume identical at baseline increased significantly in PO patients (31.7 +/- 5.3 versus 40.2 +/- 11.1%; P = 0.001) but was unchanged in PE patients (33.8 +/- 4.9 versus 34.7 +/- 6.6%; P = 0.50). It is concluded that: (1) nondiabetic nephropathy is present in approximately 15% of albuminuric type 2 diabetic patients; and (2) Perindopril prevents interstitial expansion in hypertensive patients with biopsy-proven diabetic glomerulopathy. These results support a role of angiotensin II in the progression of interstitial changes in type 2 diabetic patients with nephropathy. PMID- 10361864 TI - Acute and chronic renal effects of recombinant human TGF-beta2 in the rat. AB - The expression of transforming growth factor-beta (TGF-beta) correlates with the incidence of renal glomerular and interstitial injury, however, nothing is known of the effect of these proteins on renal hemodynamics. This study examines the renal hemodynamic and morphologic effects of recombinant human TGF-beta2 in normal male Sprague Dawley rats. Acute infusion of TGF-beta (1.2 microg/kg per min) induced no hemodynamic changes, except for a modest though significant fall in mean arterial pressure. Administering TGF-beta2 at varying doses (20, 100, and 400 microg/kg) for 9 wk caused modest increases in systolic BP and proteinuria and minimal tubular interstitial fibrosis, however, renal hemodynamic end points were not significantly altered. TGF-beta2 (800 microg/kg) was also administered to volume-depleted rats for 7 consecutive days. In contrast to the findings in volume-replete animals, administration of TGF-beta2 to volume-depleted rats caused a marked reduction in GFR and medullary blood flow. Histologic fibrosis of the medullary vasa recta and cortical interstitium was seen, but glomeruli were unaffected. Thus, acute and short-term chronic TGF-beta2 administration did not induce major renal changes in the volume-replete state, however, TGF-beta2 combined with volume depletion caused medullary hypoperfusion and reduced GFR. PMID- 10361865 TI - New phosphate binding agents: ferric compounds. AB - Several prior studies suggest that ferric compounds bind dietary phosphate and possess clinical potential as phosphate binding agents. Therefore, this study was conducted to measure the effect of several ferric compounds on intestinal phosphate binding and absorption. Balance studies lasting 2 to 4 wk were performed in normal and azotemic (achieved by subtotal nephrectomy) rats maintained on a 1.02% phosphorus diet supplemented with ferric salts (formulated to 0.95% Fe) or no ferric salt (control). In rats with normal renal function (average creatinine clearance, 4.0 ml/min per kg), the average net intestinal absorption of phosphate over all balance periods was 103.3 mg/d for the control group versus 84.7 mg/d for the ferric citrate group (P < 0.005). In the azotemic rats (average creatinine clearance, 3.3 ml/min per kg), the average net intestinal absorption of phosphate over all balance periods was significantly lower for the three ferric groups than the control groups (P < or = 0.02): 95.3 mg/d for the control group versus 75.6 mg/d for the ferric ammonium citrate treated group (P = 0.058), 77.0 mg/d for the ferric citrate-treated group (P = 0.057), and 62.5 mg/d for the ferric chloride-treated group (P < 0.002). Urinary phosphate excretion fell, sometimes to an even greater extent than did intestinal absorption, yielding no net reduction in phosphate balance in these growing, young animals with relatively preserved renal function. Calcium balance was largely unaffected by the ferric compounds. There were trends toward decreased serum phosphorus and parathyroid hormone concentrations and increased iron and hematocrit in the ferric-treated azotemic groups. All tested ferric compounds were well tolerated, but animal growth was stunted in the ferric chloride animals compared with the control group. Phosphate binding was estimated at 85 to 180 mg per gram of elemental iron, which is comparable to other phosphate binding agents. Ferric salts decrease net intestinal phosphate absorption and hold promise for the treatment of phosphate retention in patients with renal failure. PMID- 10361866 TI - Prevalence, predictors, and consequences of late nephrology referral at a tertiary care center. AB - Despite improvements in dialysis care, mortality of patients with end-stage renal disease (ESRD) remains high. One factor that has thus far received little attention, but might contribute to morbidity and mortality, is the timing of referral to the nephrologist. This study examines the hypothesis that late referral of patients to the nephrologist might lead to suboptimal pre-ESRD care. Clinical and laboratory data were obtained from the patient records and electronic databases of New England Medical Center, its affiliated dialysis unit (Dialysis Clinics, Inc., Boston), and the office records of the outpatient nephrology clinic. Early (ER) and late (LR) referral were defined by the time of first nephrology encounter greater than or less than 4 mo, respectively, before initiation of dialysis. Multivariate models were built to explore factors associated with LR, and whether LR is associated with hypoalbuminemia or late initiation of dialysis. Of the 135 patients, 30 (22%) were referred late. There were no differences in age, gender, race, and cause of ESRD between ER and LR patients. However, there were significant differences in insurance coverage between these two groups. In the multivariate analysis, patients covered by health maintenance organizations were more likely to be referred late (odds ratio = 4.5) than patients covered by Medicare. Compared to ER, LR patients were more likely to have hypoalbuminemia (56% versus 80%), hematocrit <28% (33% versus 55%), and predicted GFR <5 ml/min per 1.73 m2 (17% versus 40%) at the start of dialysis, and less likely to have received erythropoietin (40% versus 17%) or have a functioning permanent vascular access for the first hemodialysis (40% versus 4%). It is concluded that late referral to the nephrologist is common in the United States and is associated with poor pre-ESRD care. Pre-ESRD care of patients treated by nephrologists was also less than ideal. The patient-, physician-, and system-related factors behind this observation are unclear. Meanwhile, pre-ESRD educational efforts need to target patients, generalists, and nephrologists. PMID- 10361867 TI - Effects of high-dose folic acid and pyridoxine on plasma and erythrocyte sulfur amino acids in hemodialysis patients. AB - In this investigation, sulfur amino acids (sAA) and sulfhydryls were determined in the plasma and erythrocytes (RBC) of 10 uremic patients on regular hemodialysis (HD) treatment and 10 healthy subjects, before and after supplementation with 15 mg/d of folic acid and 200 mg/d of pyridoxine for 4 wk. The basal total plasma concentrations of homocysteine (Hcy), cysteine (Cys), cysteinylglycine (Cys-Gly), gamma-glutamylcysteine (gamma-Glu-Cys), glutathione (GSH), and free cysteinesulfinic acid (CSA) were significantly higher in HD patients when compared to healthy subjects, whereas methionine (Met) and taurine (Tau) concentrations were the same in the two groups. HD patients showed significantly higher RBC levels of Hcy and Cys-Gly, whereas the RBC concentrations of Met, Cys, Tau, and GSH were not different from those in the healthy subjects. The plasma concentrations of sAA and sulfhydryls differed compared with RBC levels in the healthy subjects and HD patients. In both groups, supplementation with high doses of folic acid and pyridoxine reduced the plasma Hcy concentration. In addition, increased plasma concentrations of Cys-Gly and GSH were found in the HD patients and of CSA in the healthy subjects. After vitamin supplementation, the RBC concentrations of Hcy, Cys, and GSH increased and that of Tau decreased in healthy subjects. The only significant finding in RBC of HD patients was an increase in GSH levels after supplementation. This study shows several RBC and plasma sAA and sulfhydryl abnormalities in HD patients, which confirms earlier findings that RBC and plasma pools play independent roles in interorgan amino acid transport and metabolism. Moreover, high-dose supplementation with folic acid and pyridoxine significantly reduced Hcy levels, but did not restore the sAA and sulfhydryl abnormalities to normal levels. The increase that was observed in GSH after vitamin supplementation may have a beneficial effect in improving blood antioxidant status in uremic patients. Finally, the findings of elevated plasma Cys levels correlating to the elevated plasma Hcy levels in the presence of elevated plasma CSA levels, both before and after vitamin supplementation, led to the hypothesis that a block in decarboxylation of CSA is linked to hyperhomocysteinemia in end-stage renal failure. PMID- 10361868 TI - QT dispersion in patients with end-stage renal failure and during hemodialysis. AB - Interlead variability of the QT interval in surface electrocardiogram (ECG), i.e., QT dispersion, reflects regional differences in ventricular recovery time, and it has been linked to the occurrence of malignant arrhythmias in different cardiac diseases. The purpose of the study was to assess the effect of hemodialysis on QT and corrected QT (QTc) interval and dispersion in chronic hemodialyzed patients. Data of 34 nondiabetic patients (male/female = 21/13; mean age, 54 +/- 15 yr) on chronic hemodialysis were studied. Polysulfone capillaries and bicarbonate dialysate containing (in mEq/L) 135 Na+, 2.0 K+, 1.5 Ca2+, and 1.0 Mg2+ were used. Simultaneous 12-lead ECG were recorded before and after hemodialysis in a standard setting. The QT intervals for each lead were measured manually on enlarged (x3) ECG by one observer using calipers. Each QT interval was corrected for patient heart rate: QTc = QT/square root of RR (in milliseconds [ms]). The average cycle intervals were 853 +/- 152 ms predialysis and 830 +/- 173 ms postdialysis; the difference was not significant. The maximal QT interval changed significantly from 449 +/- 43 to 469 +/- 41 ms (P < 0.01). The corrected maximal QT interval increased significantly from 482 +/- 42 to 519 +/- 33 ms (P < 0.01). The QT dispersion changed from 56 +/- 15 to 85 +/- 12 ms (P < 0.001) and the corrected QT interval dispersion from 62 +/- 18 to 95 +/- 17 ms (P < 0.001). During hemodialysis, the serum potassium and phosphate levels decreased from 5.5 +/- 0.8 to 3.9 +/- 0.5 (mM) and from 2.3 +/- 0.5 to 1.6 +/- 0.4 (mM), respectively, whereas calcium increased from 2.2 +/- 0.23 to 2.5 +/- 0.22 (mM). It is concluded that hemodialysis increases the QT and QTc interval and QT and QTc dispersion in patients with end-stage renal failure. Thus, it may be stated that the nonhomogeneity of regional ventricular repolarization increases during hemodialysis. Measurement of QT and QTc dispersion is a simple bedside method that can be used for analyzing ventricular repolarization during hemodialysis. PMID- 10361869 TI - Effect of intravenous saline, albumin, or hydroxyethylstarch on blood volume during combined ultrafiltration and hemodialysis. AB - It is generally advocated to use saline or albumin infusions during symptomatic hypotension during dialysis. However, because of their side effects and/or costs, they are of limited use. Hydroxyethylstarch (HES), a synthetic colloid with a long-standing volume effect, is used in the management of hypovolemia. In this study, the efficacy of three fluids (isotonic saline [0.9%], albumin [20%], and HES [10%]) was assessed during three treatment sessions with combined ultrafiltration and hemodialysis, which differed in the type of fluid given intravenously. Changes in relative blood volume (BV), systolic BP (SBP), and vascular reactivity (venous tone [VT]) were compared. An intravenous infusion of 100 ml of fluid was given when the decrease in BV versus baseline was more than 10% as measured by a continuous optical reflection method. The ultrafiltration was continued. BV decreased significantly versus baseline independent of the intravenous fluid administration in all three treatment sessions. However, when we compared BV values at the end of the dialysis session with those at the time of infusion, BV continued to decrease significantly with saline (change in BV 4.56 +/- 2.75%; P < 0.05) and albumin (change in BV -2.13 +/- 2.51%; P < 0.05), but not with HES (change in BV -0.15 +/- 2.17%; NS). Between albumin and HES there were no significant differences in changes in BV (NS), whereas between HES and saline (P < 0.05) and between albumin and saline (P < 0.05) the differences in BV changes were significant. SBP remained unchanged within each session. Although SBP tended to decrease more with saline compared to albumin and HES, the difference was not significant. The higher decrease in BV and SBP with saline was counterbalanced by a significantly higher increase in VT, while VT remained unchanged in the other two sessions. It is concluded that HES is a promising fluid in preserving blood volume, comparable to albumin, but superior to saline. PMID- 10361870 TI - Hematocrit levels and hospitalization risks in hemodialysis patients. AB - The association between hematocrit level and future hospitalization risks in hemodialysis patients has not been fully investigated on a national level. A total of 71,717 prevalent Medicare hemodialysis patients who survived a 6-mo entry period from July 1 through December 31, 1993 were studied, and their risk of hospitalizations was evaluated the next year. Five hematocrit groups were defined from Medicare recombinant human erythropoietin-treated patients: <27%, 27 to <30%, 30 to <33%, 33 to <36%, and > or =36%. A Cox regression model was used to investigate the association between hematocrit level and the risk of first hospitalization, and the Andersen-Gill regression model evaluated multiple hospitalizations during the next year, adjusting for patient comorbidity and severity of disease. Compared with the baseline group of 30 to <33%, patients with hematocrit levels <30% had a 14 to 30% increased risk of hospitalization without disease severity adjustment (p = 0.0001) and a 7 to 18% increased risk with disease severity adjustment (p = 0.0001). Patients in the 33 to <36% group had the lowest risk at 0.93 and 0.88 (p = 0.0001), with and without adjustment for disease severity. It is concluded that patients with hematocrits of <30% have an increased risk of future hospitalization, with hematocrit levels between 33 and 36% having the lowest associated risks. PMID- 10361871 TI - Influence of donor factors on early function of graft kidneys. AB - Factors which influence graft function can be divided into donor factors that affect both kidneys from the same donor equally and postdonor factors that affect each kidney individually. This study assessed the influence of donor factors on graft function early after transplantation. Sixty-one donors who provided kidneys that were transplanted locally into two separate recipients were identified. Recipient creatinine clearance values were estimated from serum creatinine concentrations using a computer model. Pairwise ANOVA showed that donor factors accounted for 35 to 45% of the variation in recipient creatinine clearance from 2 d to 2 wk posttransplantation. Although donor factors had a large aggregate effect during this period, individual factors that influenced graft function could not be identified from analysis of donor medical records. At 6 mo after transplantation, the effect of donor factors on graft function was no longer discernible. These results show that the condition of the donor exerts an important influence on graft function early after transplantation. More detailed study is required to identify individual factors that contribute to this effect. PMID- 10361872 TI - Antihypertensive medication and renal allograft failure: a North American Pediatric Renal Transplant Cooperative Study report. AB - Hypertension after renal transplantation occurs commonly and, in adults, is associated with decreased graft survival. The North American Pediatric Renal Transplant Cooperative Study database was analyzed to determine: (1) the percent use of antihypertensive (anti-HTN) medication based on donor type, race, age, and acute rejection status; and (2) whether use of anti-HTN medication is associated with higher rates of subsequent graft failure. Data regarding anti-HTN medication use was available in 5251 renal allografts (4821 patients) with >30 d graft function. Posttransplant follow-up data were collected at 30 d, 6 mo, 12 mo, and then annually for 5 yr. At each follow-up, patients were selected for further analysis if the graft was functioning at that visit and subsequent follow-up data were available. Overall, anti-HTN medication use was 79% on day 30 and 58% at 5 yr. At each follow-up, anti-HTN medication use was higher (P < 0.01) for cadaveric donor versus living related donor, blacks versus whites, age >12 versus <12 yr, and > or = 1 versus 0 acute rejection episodes. Anti-HTN medication use at each annual follow-up was associated with significantly higher rates of subsequent graft failure. Multiple regression analysis controlling for all factors associated with increased use of anti-HTN medications revealed a relative risk of graft failure for use of anti-HTN medication of greater than 1.4 (P < 0.001). In recipients of cadaveric allografts, only acute rejection status predicted subsequent graft failure more strongly than use of anti-HTN medications. These data suggest that hypertension after renal transplantation in children, as evidenced by use of anti-HTN medications, is associated with increased rates of subsequent graft failure. PMID- 10361874 TI - Transmembrane proteins in the tight junction barrier. AB - Three types of transmembrane proteins have been identified within the tight junction, but it remains to be determined how they provide the molecular basis for regulating the paracellular permeability for water, solutes, and immune cells. Several of these proteins localize specifically within the continuous cell to-cell contacts of the tight junction. One of these, occludin, is a cell adhesion molecule that has been demonstrated to influence ion and solute permeability. The claudins are a family of four-membrane spanning proteins; unexpectedly, other members of this family have already been characterized without recognizing their relationship to tight junctions. Junction adhesion molecule, the most recently identified tight junction component, is a member of the Ig superfamily and influences the paracellular transmigration of immune cells. A plaque of cytoplasmic proteins under the junction may be responsible for scaffolding the transmembrane proteins, creating a link to the perijunctional actin cytoskeleton and transducing regulatory signals that control the paracellular barrier. PMID- 10361873 TI - Effect of danaparoid sodium on proteinuria, von Willebrand factor, and hard exudates in patients with diabetes mellitus type 2. AB - In diabetic nephropathy, heparan sulfate glycosaminoglycan side chains are reduced in glomerular basement membranes proportionally to the degree of proteinuria. Recently, it was demonstrated that additional therapy with danaparoid sodium, a mixture of sulfated glycosaminoglycans with mainly heparan sulfate, lowered proteinuria in type 1 diabetes patients with diabetic nephropathy. A randomized placebo-controlled parallel study was performed with 750 anti-Xa units of danaparoid sodium once daily in type 2 diabetes patients with severe proteinuria. The aim of the study was to evaluate the possible effects of danaparoid sodium on proteinuria, endothelial dysfunction, and hard exudates in the retina and to determine the safety/tolerability of this drug. Twenty-two patients completed the study, and one patient had to stop prematurely after 6 wk of danaparoid sodium treatment because of urticaria at the injection sites. Apart from a small decrease of hemoglobin and minor skin hematomas at the injection site in five patients in the danaparoid sodium group, no other safety parameters showed any clinically or statistically significant difference between and within groups. The relative change in time of both the urinary albumin and protein excretion rate corrected for creatinine did not differ between both treatment arms (P = 0.2 and 0.49, respectively). No retinal complications or changes of hard exudates occurred. von Willebrand factor was elevated in both groups, but was not influenced by either treatment modality. Contrary to the beneficial effects that occurred in type 1 diabetes patients with diabetic nephropathy, treatment for 8 wk with 750 anti-Xa units of danaparoid sodium gave no reduction of proteinuria, hard exudates, and von Willebrand factor. PMID- 10361875 TI - HLA-derived peptides as novel immunomodulatory therapeutics. AB - A growing body of experimental evidence demonstrates that synthetic peptides corresponding to linear sequences of MHC (HLA in humans) proteins have immunomodulatory effects in vitro and in vivo in animal models and in humans. Although the original concept was that these peptides inhibited antigen recognition at the MHC-T cell receptor interface via physical blockade, it is now clear that the mechanisms responsible for the myriad of functional effects are more complex. Recent findings show that some peptides affect signal transduction and cell cycle progression. Fragments of MHC molecules can dampen or downregulate immune responses via a variety of mechanisms. Some soluble MHC molecules or synthetic peptides are capable of inducing and maintaining immunologic tolerance in animals. This information suggests that synthetic peptides themselves or drugs mimicking their effects may represent a new class of immunotherapeutics. PMID- 10361876 TI - T cell costimulatory blockade: new therapies for transplant rejection. AB - Optimal T cell responses occur when T cells receive both antigen-specific signals through the T cell receptor and non-antigen-specific costimulatory signals through accessory cell surface molecules. The best understood costimulatory receptor is CD28. Signals through the T cell receptor and CD28 cooperatively induce cytokine gene expression and promote T cell proliferation and survival. Negative signals delivered through a related cell surface receptor, cytotoxic T lymphocyte antigen (CTLA-4), act to terminate immune responses and are required for normal immune homeostasis. This article reviews T cell costimulation, including the CD28/CTLA-4 system and other potential costimulatory pathways (such as CD40/CD154), the role of these pathways in normal immune responses, and the potential for the inhibition of these pathways to induce transplantation tolerance. PMID- 10361877 TI - Newer immunosuppressive drugs: a review. AB - In recent years, many new immunosuppressive drugs have been discovered and developed for clinical use in transplantation. This review focuses on those drugs (leflunomide, mycophenolate mofetil, sirolimus, tacrolimus) that have been shown to have immunosuppressive activity in patients. Different anti-interleukin-2 receptor antibodies are also reviewed as an example of a resurgence of development in the area of monoclonal antibodies. The price for reducing the incidence of allograft rejection by improved immunosuppression was thought to be a proportional increase in the incidence of infection and malignancy. Data from Phase III clinical trials of new immunosuppressants, however, show a statistically significant reduction in the incidence of acute rejection produced by these new drugs, which has not been accompanied by increases in infection and malignancy rates. The wide array of new drugs offers the opportunity to use combinations that block different pathways of immune activation while at the same time selecting drug combinations with nonoverlapping toxicity profiles so that doses of each single drug can be reduced below toxicity levels. The immunosuppressive therapy for patients can be tailored according to their individual needs. PMID- 10361878 TI - Sexual dysfunction in uremia. AB - In summary, sexual dysfunction is a common finding in both men and women with chronic renal failure. Common disturbances include erectile dysfunction in men, menstrual abnormalities in women, and decreased libido and fertility in both sexes. These abnormalities are primarily organic in nature and are related to uremia as well as the other comorbid conditions that frequently accompany the chronic renal failure patient. Fatigue and psychosocial factors related to the presence of a chronic disease are also contributory factors. Disturbances in the hypothalamic-pituitary-gonadal axis can be detected before the need for dialysis but continue to worsen once dialytic therapy is initiated. Impaired gonadal function is prominent in uremic men, whereas the disturbances in the hypothalamicpituitary axis are more subtle. By contrast, central disturbances are more prominent in uremic women. Therapy is initially directed toward optimizing the delivery of dialysis, correcting anemia with recombinant erythropoietin, and controlling the degree of secondary hyperparathyroidism with vitamin D. For many practicing nephrologists, sildenafil has become the first-line therapy in the treatment of impotence. In the hypogonadal man whose only complaint is decreased libido, testosterone may be of benefit. Regular gynecologic follow-up is required in uremic women to guard against potential complications of unopposed estrogen effect. Uremic women should be advised against pregnancy while on dialysis. Successful transplantation is the most effective means of restoring normal sexual function in both men and women with chronic renal failure. PMID- 10361879 TI - The role of anti-glomerular basement membrane antibody in the pathogenesis of human glomerulonephritis. PMID- 10361880 TI - Pharmacology and cardiovascular implications of the kinin-kallikrein system. AB - Kinins are peptide hormones that can exert a significant influence on the regulation of blood pressure and vascular tone due to their vasodilatatory, natriuretic and growth modulating activity. Their cardiovascular involvement in physiological and pathophysiological situations has been studied intensively since inhibitors for angiotensin I-converting enzyme and selective receptor antagonists have become available for pharmacologically potentiating or inhibiting kinin-mediated reactions. Molecular biological analysis and the establishment of genetically modified animal models have also allowed newer information to be acquired on this subject. In this review, the components and cardiovascularly relevant mechanisms of the kinin-kallikrein system shall be described. Organ-specific effects concerning the kidneys, the vascular system, the heart and nervous tissue shall also be illustrated. On this issue, the physiological functions and pathophysiological implications of the kinin kallikrein system should be clearly distinguished from the many, mostly endothelium-mediated protective effects which occur during ACE inhibition due to the potentiation of kinin effects. Finally, a view shall also be cast upon newly discovered targets of action, which could be exploited for therapeutically altering the kinin-kallikrein system. PMID- 10361881 TI - Recent advances in neuropharmacology of cutaneous nociceptors. AB - Cutaneous nociceptors are peripheral receptive endings of primary sensory neurons activated by noxious stimuli. Nociceptors detect and signal the presence of tissue-damaging stimuli or the existence of tissue damage. In this short review, we will focus on the molecular mechanism of maintenance, activation, inhibition and sensitization in cutaneous nociceptors. Neurotrophic factors are essential to the development of nociceptors during embryogenesis. Recent evidences have indicated that nociceptors in the adult are maintained by either nerve growth factor (NGF) or glial cell line-derived neurotrophic factor (GDNF). A selective activator of nociceptors is capsaicin, natural product of capsicum peppers. Recently, the receptor for capsaicin (the vanilloid receptor 1: VR1) has been cloned, identified and characterized. VR1 seems to play an important role in the activation and sensitization of nociceptors. In contrast, peripheral endogenous cannabinoids such as anandamide are novel candidates for mediators that inhibit the excitation of nociceptors. Intracellular messengers and the mechanisms of signal transduction in nociceptors have also been studied. Our recent findings provide evidences demonstrate that an activation of both cAMP- and cGMP-second messenger systems is required to induce the sensitization of nociceptors. Such emerging evidences reviewed here would make a significant contribution to further understanding of the molecular mechanism of nociceptors. PMID- 10361882 TI - Alendronate induces antinociception in mice, not related with its effects in bone. AB - The antinociceptive effect of alendronate was studied. The bisphosphonate was i.p. administered and two tests were carried out: acetic acid in mice and formalin test in rats. In the acetic acid test, alendronate induced a dose dependent antinociceptive effect that was statistically significant for the doses of 10, 20 and 40 mg/kg, and could be detected 48 hr after its administration. In the formalin test, however, alendronate, at the doses of 10 and 20 mg/kg, did not modify the pain score nor the number of flinches, when it was administered either 30 or 60 min before the test. However it must be noted that doses inducing analgesic effect are close to those inducing toxicity. PMID- 10361883 TI - Effect of a novel non-steroidal anti-inflammatory drug (M-5011) on cytokine levels in rats with monosodium urate crystal- induced pleurisy. AB - We evaluated the effects of a new non-steroidal anti-inflammatory drug (NSAID), d 2-[4-(3-methyl-2-thienyl)phenyl]propionic acid (M-5011), and indomethacin on the production of arachidonate metabolites and pro-inflammatory cytokines in male Sprague-Dawley rats with monosodium urate crystal (MSU)-induced pleurisy. Levels of tumor necrosis factor (TNF), interleukin (IL)-1 and IL-6 in the pleural exudate were determined by biological assays, while prostaglandin E2 (PGE2), leukotriene B4 (LTB4) and cytokine-induced chemoattractant-1 (CINC-1) levels were quantified by enzyme immunoassays. Orally administered M-5011 (5 mg/kg) decreased the pleural exudate volume at 3 and 4 hr after MSU injection. Indomethacin (10 mg/kg) decreased the volume at 3-5 hr. These drugs reduced the number of leukocytes in the pleural cavity at 6 hr. Both NSAIDs also reduced the content of PGE2 in the exudate without affecting LTB4 levels. Increased productions of both IL-6 and CINC-1 in the exudate were reduced by pretreatment with M-5011 or indomethacin, and TNF levels in the exudate were increased by pretreatment of these drugs. Thus, M-5011 inhibits the production of both IL-6 and CINC-1 at lower doses than those of indomethacin, and the inhibitory effect of M-5011 on CINC-1, but not IL-6, may partly contribute to the inhibition of leukocyte infiltration in rats with MSU-induced pleurisy. PMID- 10361884 TI - Naftopidil, a novel alpha1-adrenoceptor antagonist, displays selective inhibition of canine prostatic pressure and high affinity binding to cloned human alpha1 adrenoceptors. AB - The pharmacological profiles of the alpha1-adrenoceptor antagonists naftopidil, tamsulosin and prazosin were studied in an anesthetized dog model that allowed the simultaneous assessment of their antagonist potency against phenylephrine mediated increases in prostatic pressure and mean blood pressure. The intravenous administration of each of these compounds dose-dependently inhibited phenylephrine-induced increases in prostatic pressure and mean blood pressure. To further assess the ability of the three compounds to inhibit phenylephrine induced responses, the doses required to produce a 50% inhibition of the phenylephrine-induced increases in prostatic and mean blood pressure and the selectivity index obtained from the ratio of those two doses were determined for each test compound. Forty minutes after the intravenous administration of naftopidil, the selectivity index was 3.76, and those of tamsulosin and prazosin were 1.23 and 0.61, respectively. These findings demonstrated that naftopidil selectively inhibited the phenylephrine-induced increase in prostatic pressure compared with mean blood pressure in the anesthetized dog model. The selectivity of naftopidil for prostatic pressure was the most potent among the test compounds. In addition, using cloned human alpha1-adrenoceptor subtypes, naftopidil was selective for the alpha1d-adrenoceptor with approximately 3- and 17-fold higher affinity than for the alpha1a- and alpha1b-adrenoceptor subtypes, respectively. The selectivity of naftopidil for prostatic pressure may be attributable to its high binding affinity for alpha1a- and alpha1d-adrenoceptor subtypes. PMID- 10361885 TI - Angiotensin II type 1 receptor antagonist, TCV-116, prevents neointima formation in injured arteries in the dog. AB - We investigated the effect of an angiotensin (Ang) II antagonist, (+/-)-1 (cyclohexyloxycarbonyloxy)-ethyl 2-ethoxy- 1- [[2'-(1H-tetrazol-5-yl)biphenyl-4 yl]methyl]- 1H-benzimidazole-7-carboxylate (TCV-116), on neointima formation in dog artery injured by a balloon catheter. Dogs were orally treated with 10 mg/kg TCV-116 or placebo twice a day for 5 weeks. After treatment with these drugs for 1 week, the right carotid artery was injured by a balloon catheter. The left carotid artery was regarded as the control. In the group treated with placebo, neointima formation in the injured arteries was observed. The activities of angiotensin converting enzyme (ACE) and chymase in the injured carotid arteries were increased 2.56- and 3.26-fold compared with those in the non-injured arteries, respectively. The neointimal area in dogs treated with placebo and TCV 116 were 0.51 +/-0.07 and 0.21 +/-0.07 mm2, respectively, and this difference was significant. In conclusion, an Ang II antagonist, TCV-116, prevented neointima formation by blocking the action of Ang II generated by both ACE and chymase in the injured arteries. PMID- 10361886 TI - Effects of imipramine, an uptake inhibitor, on double-peaked constrictor responses to periarterial nerve stimulation in isolated, perfused canine splenic arteries. AB - Using a cannula insertion method, periarterial nerve electrical stimulations were performed at 1 and 10 Hz in the isolated, perfused canine splenic artery. Electrical nerve stimulation readily caused double-peaked vasoconstrictions. The 1st-peak response at 1 Hz was not influenced by treatment with imipramine but the 2nd one was significantly enhanced by it. The 2nd-peak response was markedly blocked by prazosin. An additional treatment with alpha,beta-methylene ATP, a P2X purinoceptor desensitizer, abolished electrical stimulation-induced vascular responses that remained. At 10 Hz, the responses to electrical stimulation were not significantly influenced by imipramine. On the other hand, the imipramine treatment inhibited the tyramine-induced vasoconstriction but potentiated the noradrenaline-induced one. ATP-induced responses were not modified by imipramine. From these results, it is concluded that 1) the 1st-peaked constriction is mainly due to a P2X-purinoceptor-dependent mechanism, 2) the 2nd one is mainly due to an alpha1-adrenoceptor-dependent mechanism, and 3) presynaptic uptake mechanisms may perform an important role in the regulation of vascular reactivity, especially at a low frequency. PMID- 10361887 TI - Bacterial expression and functional characterization of a rat thyroid hormone sulfotransferase, ST1B1. AB - At least three forms of phenol sulfotransferase (ST) ST1B1, ST1A1 and ST1C1 are contained in rat livers. To identify the form contributing to the metabolism of 3,3',5-triiodothyronine (T3), functional characterization of these forms was performed by expression in Escherichia coli. ST1B1 and ST1C1 were shown to be active on sulfation towards T3 with high affinity (Km: 44.4 and 25.8 microM, respectively), whereas ST1A1 had low affinity. In Western blotting using antibodies raised against the individual ST, hepatic contents of each ST were quantitatively determined. ST1B1 showed no clear sex-difference, whereas the level of ST1C1 was higher in adult males than adult females. The content of ST1B1 was 1.4, 6.8 and 10 times higher than that of ST1C1 in adult males, adult females and both sexes of immature rats, respectively. The developmental pattern of ST1B1 was similar to that of ST1A1, but differed from that of ST1C1. These results indicate that ST1B1 and ST1C1 are involved in T3 metabolism in rats and ST1B1 is the constitutive form across sexes and ages. PMID- 10361889 TI - The protective effect of cyclosporine A on anti-Fas antibody-induced hepatitis in mice. AB - The effect of cyclosporine A (CsA) on anti-Fas antibody-induced hepatitis was studied. The administration of anti-Fas antibody (250 microg/kg) to mice elevated plasma alanine aminotransferase (ALT) activity at 3 hr. This anti-Fas antibody induced elevation of ALT was inhibited by treatment with CsA (10, 30 and 100 mg/kg) in a dose-dependent manner. Anti-Fas antibody administration elevated CPP32-like protease activity at 3 hr in mouse liver, and this elevation of CPP32 like activity was inhibited by treatment with CsA. The present results show that CsA treatment inhibits the anti-Fas antibody-induced apoptotic process of hepatitis, at least in part, by affecting a reaction upstream of CPP32-like protease activation. PMID- 10361888 TI - Effects of XT-44, a phosphodiesterase 4 inhibitor, in osteoblastgenesis and osteoclastgenesis in culture and its therapeutic effects in rat osteopenia models. AB - We have reported that denbufylline, a phosphodiesterase 4 (PDE4) inhibitor, inhibits bone loss in Walker256/S tumor-bearing rats, suggesting therapeutic potentiality of a PDE4 inhibitor in osteopenia. In the present study, effects of a new PDE4 inhibitor, 1-n-butyl-3-n-propylxanthine (XT-44), in bone were evaluated in cell cultures and animal experiments. In rat bone marrow culture, XT 44 stimulated mineralized-nodule formation, whereas it inhibited osteoclast-like cell formation in mouse bone marrow culture. In Walker256/S-bearing rats (6-week old female Wistar Imamichi rats), rapid decrease in bone mineral density (BMD) was prominent, and oral administration of XT-44 (0.3 mg/kg, every 2 days) inhibited the decrease in BMD. In the second animal experiment, female Wistar rats (6-week-old) were sciatic neurectomized, and XT-44 was orally administered to these rats every 2 days for 4 weeks. XT-44 administration (0.3 mg/kg) recovered BMD in these neurectomized animals. Furthermore, 19-week-old, female Wistar rats were ovariectomized (OVX), and 15 weeks after surgery, these rats were orally administered XT-44 every 2 days for 8 weeks. XT-44 treatment (1 mg/kg) increased the BMD of OVX rats. These results indicate that XT-44 could be a candidate as a therapeutic drug for treating osteopenia including osteoporosis. PMID- 10361890 TI - Inhibitory effects of Sasa senanensis Rehder extract (SE) on calcium-ionophore A23187-induced histamine release from rat peritoneal exudate cells. AB - The effects of Sasa senanensis Rehder extract (SE) by alkaline hydrolysis on histamine release from rat peritoneal exudate cells (PECs) were examined. Preincubation with SE for 5 min suppressed calcium-ionophore A23187-induced histamine release in a concentration-dependent manner. A23187 evoked a quick increase in cytoplasmic free calcium ([Ca2+]i) levels in the presence of extracellular calcium. Preincubation with SE also had an inhibitory effect on calcium influx increases induced by A23187. These results indicate that SE prevents degranulation from rat PECs by inhibiting [Ca2+]i level elevation. PMID- 10361891 TI - Ability of mosapride to bind to 5-HT4 receptor in the human stomach. AB - Ability of mosapride, a gastrokinetic agent, to bind to 5-HT4 receptor was examined in the stomach of human and guinea pig by in vitro receptor autoradiography. [125I]SB207710 binding sites were detected in the muscle layer including the myenteric plexus of the stomach from both humans and guinea pigs, although the binding was observed more clearly and densely in the stomach of guinea pigs than humans. Mosapride as well as SB204070 inhibited the binding of [125I]SB207710. Thus, mosapride possesses the ability to bind to 5-HT4 receptors of human stomach and may modulate the motility, as in the case of guinea pig stomach. PMID- 10361892 TI - Studies on the anxiolytic activity of Eurycoma longifolia Jack roots in mice. AB - The anxiolytic effect of Eurycoma longifolia Jack in mice was examined. Fractions of E. longifolia Jack extract produced a significant increase in the number of squares crossed (controls= 118.2 +/- 10.2 squares), but significantly decreased both the immobility (controls = 39.4+/- 4.0 sec) and fecal pellets (controls= 12.3 +/-2.1 fecal pellets) when compared with control mice in the open-field test; they significantly increased the number of entries (controls=6.7+/-0.5 entries) and time spent (controls=42.9+/-0.1 sec) in the open arms, but decreased both the number of entries (controls= 13.2+/-0.7 entries) and time spent (controls= 193.4+/-0.7 sec) when compared with the control mice in the closed arms of the elevated plus-maze test. Furthermore, fractions of E. longifolia Jack extract decreased the fighting episodes significantly (controls= 18.0+/-0.4 fighting episodes) when compared with control mice. In addition, these results were found to be consistent with anxiolytic effect produced by diazepam. Hence, this study supports the medicinal use of this plant for anxiety therapy. PMID- 10361893 TI - IPD-1151T (suplatast tosilate) inhibits interleukin (IL)-13 release but not IL-4 release from basophils. AB - The effect of suplatast tosilate (IPD-1151T), which is known to suppress interleukin (IL)-4 release from T cells, on the release of IL-4 and IL-13 from human peripheral basophils was investigated. Basophils were obtained from 16 mite sensitive atopic asthmatic patients. IPD-1151T clearly inhibited the antigen induced release of IL-13 but not IL-4. These results suggest that IPD-1151T possesses different activity for the regulation of cytokine release in basophils and T cells. PMID- 10361894 TI - Activation of in vivo Kupffer cell function by oral administration of Cordyceps sinensis in rats. AB - We investigated the effect of water extracts of Cordyceps sinensis (WECS) on Kupffer cell function in rats. Rats were received a single i.v. injection of a colloidal carbon solution and then the clearance rate from the blood were measured. The rats had been daily administered with WECS, p.o. at a dose of 200 mg/kg for 25 days until the day before the injection of colloidal carbon. The half-life of the colloidal carbon in the blood of rats administered WECS 200 mg/kg was significantly shorter than that of the control rats. This suggests that accelerated function of Kupffer cells is partially involved in the anti metastatic action of WECS. PMID- 10361895 TI - Fecal occult blood tests in occult gastrointestinal bleeding. AB - Occult gastrointestinal bleeding is diagnosed by using one of the commercially available fecal occult blood tests (FOBTs). Guaiac-based slide tests are most frequently used, although the more specific immunochemical methods are promising. The guaiac tests are inexpensive, nonspecific, qualitative measures of stool blood, and their use requires dietary and drug restrictions. Clinicians need to be aware of the causes of false-positive and false-negative test results. Although specific for the presence of human blood, immunochemical tests are more expensive and tend to react also to physiological quantities of blood in fecal specimens. Whichever test is chosen, it must be processed and read correctly. Annual FOBT screening for colorectal cancer, combined with periodic flexible sigmoidoscopy, is a cost-effective method of detecting early, curable colorectal cancer. PMID- 10361896 TI - Gastrointestinal tract evaluation in patients with iron deficiency anemia. AB - Iron deficiency anemia is the most common form of anemia encountered in clinical practice and is an extremely common manifestation of chronic occult gastrointestinal bleeding. Current evidence suggests that a large proportion of men and postmenopausal women with iron deficiency anemia harbor significant gastrointestinal tract pathological lesions as the source of blood loss. As such, the evaluation of patients with iron deficiency anemia is generally focused on the gastrointestinal tract. Importantly, the diagnosis of iron deficiency anemia should be firmly established before an extensive evaluation is undertaken. Management strategies for patients with iron deficiency anemia are reviewed; an important general point is that clinical features (ie, symptoms) may help direct specific investigation. The role of small-intestinal investigation in patients with iron deficiency anemia is controversial and should probably be reserved for patients with iron deficiency anemia and persistent gastrointestinal symptoms or those who fail to respond to appropriate therapy. The treatment and prognosis of patients with iron deficiency anemia and the majority of gastrointestinal tract lesions are straightforward. However, patients with vascular ectasias as the source of blood loss can represent a true management challenge. PMID- 10361897 TI - Small-bowel investigation in occult gastrointestinal bleeding. AB - Obscure gastrointestinal bleeding after careful endoscopy of the upper and lower gastrointestinal tract is predominantly of small-bowel origin. Patients presenting with overt blood loss account for a select subpopulation of those with small-bowel bleeding. Although relatively rare, these patients often require repeated blood transfusions, investigation, and hospitalization before a diagnosis is reached. These events have a considerable negative impact on the patient's quality of life. Standard evaluation using enteroclysis, tagged red cell studies, and angiography are proven to be of limited value in this context. Push enteroscopy has significant advantages in this patient group, with the ability to deliver endoscopic therapy. Sonde enteroscopy is now reserved for a few patients to guide decisions on surgery, particularly in those with significant medical comorbidity. Definitive evaluation may require perioperative enteroscopy, but many patients can be managed without the need for surgery. A team approach by physician, radiologist, and surgeon following locally agreed algorithms is essential for the successful management of this challenging clinical problem. PMID- 10361898 TI - Medical and hormonal therapy in occult gastrointestinal bleeding. AB - In this age of modern technology and aggressive but noninvasive therapies, the idea of treating an identifiable but discrete bleeding lesion with systemic medical therapy seems an anachronism. But medical therapy can be the treatment of choice for some bleeding vascular lesions of the gut. Though most vascular lesions appear similar endoscopically and are a cause of gastrointestinal bleeding, they consist of various pathologic identities. These different lesions have not only different pathologic appearances, but also different prognoses. The natural history of many of these lesions remains largely unknown. Long-term success in controlling bleeding must be measured in the context of the responsible lesion's frequency of occurrence and recurrence. Medical therapy can include hopeful watchful waiting, routine blood transfusions, or specific medications. Medical therapy has been pursued along two lines. The most common form of medical therapy has been simple supportive care. This may include iron therapy and avoidance of aspirin and other anticoagulants. Transfusions may be necessary, occasionally or on a regular basis. The second form of medical therapy has been the use of estrogens. There have been other medical attempts to control bleeding from intestinal vascular lesions. Somatostatin has been used in an uncontrolled fashion, as has aminocaproic acid. Vascular lesions of the bowel are not all the same. Medical therapy of vascular lesions is contrary to general present practice. Endoscopic or surgical therapy is presently considered best because of its ease, relatively good long-term results, and the lack of a clearly effective, well-tolerated medical therapy. Medical therapy is usually reserved for diffuse vascular diseases of the bowel, for vascular lesions located in relatively inaccessible locations, for patients with continued bleeding despite endoscopic or surgical management, and for patients who are not candidates for either endoscopic or surgical therapy. PMID- 10361899 TI - The role of surgery in occult gastrointestinal bleeding. AB - The surgeon is frequently involved in the management of patients with occult gastrointestinal bleeding. It is important to have a systematic approach to these patients to avoid the "looking for a needle in a haystack" approach to this problem. These are a group of patients who have undergone extensive standard gastroendoscopic evaluation and continue to bleed. Five percent of gastrointestinal bleeding occurs between the ligament of Treitz and the ileocecal valve. Therapeutic management may be guided by the age of the patient. Patients aged younger than 50 years will usually bleed from readily identifiable palpable lesions, such as leiomyoma, Meckel's diverticulum, or other small-bowel tumors, whereas the patients aged older than 50 years most commonly bleed from angiodysplasias or arteriovenous malformations that are not palpable, frequently multiple, and may be evanescent. PMID- 10361900 TI - It is the time to ask what adenosine can do for cardioprotection in ischemic heart disease. PMID- 10361901 TI - A new approach for the treatment of acute porphyria. PMID- 10361902 TI - Carbonyl stress in the pathogenesis of diabetic nephropathy. AB - Diabetic nephropathy is a major chronic complication of diabetes mellitus and an important cause of increased morbidity and mortality in diabetic patients. Although several lines of evidence have suggested that poor glycemic control undoubtedly plays a significant role, the metabolic events responsible for its development are not understood well. Possible mediators of untowards effects of hyperglycemia include the advanced glycation end products (AGEs). AGEs, carboxymethyllysine and pentosidine, whose formation is closely linked to oxidation, accumulate in the characteristic diabetic glomerular lesions, such as the expanded mesangial matrix and nodular lesions, in co-localization with other oxidation-specific protein adducts, such as malondialdehyde-lysine, 4 hydroxynonenal-protein adduct, and acrolein-protein adduct. These five biomarkers are formed under oxidative stress by carbonyl amine chemistry between protein amino group and carbonyl compounds derived from carbohydrates, lipids, and amino acids. This article focuses on new aspects of the pathology of diabetic nephropathy, implicating an increased oxidative stress and carbonyl modification of proteins by autoxidation products of carbohydrates, lipids, and amino acids in diabetic glomerular tissue damage ("carbonyl stress"). PMID- 10361903 TI - What have we learned from gene targeting studies for the renin angiotensin system of the kidney? AB - Over the last decade, gene targeting technologies have provided investigators with powerful new tools to study the physiology and pathophysiology of the kidney. In that, the renin-angiotensin system (RAS) has been a subject of intense investigation. Detailed analyses of mutant mice have not only confirmed notions already suggested by other studies, but also shed a new light on previously unrecognized functions of RAS. In this review, we will focus on what we have learned from these gene targeted animals in particular relevance to nephrology. PMID- 10361904 TI - Adenosine 5'-triphosphate induced dilation of human coronary microvessels in vivo. AB - OBJECT: This study was performed to compare the coronary microvascular response to adenosine 5'-triphosphate (ATP) with the response to adenosine in humans. METHODS: Coronary blood flow velocity was determined using a Doppler flow wire. After intracoronary nitroglycerin infusion, intracoronary bolus injections of adenosine (20 microg) and ATP (20 microg) were performed to induce reactive hyperemia. PATIENTS: Twenty-nine patients (23 men and 6 women, mean age: 63+/-9 years) with coronary artery disease and risk factors for coronary atherosclerosis were studied. RESULTS: Coronary flow reserve in response to ATP was similar to that for adenosine (2.7+/-0.7 vs. 2.7+/-0.7). However, the duration of ATP induced vasodilation was longer than that of adenosine-induced dilation (39+/-25 seconds vs. 26+/-12 seconds, p<0.0001). The coronary flow reserve obtained with either ATP or adenosine was significantly reduced in the interventioned arteries compared with non-stenosed arteries. The coronary flow reserve obtained with ATP was similar to that obtained with adenosine in both artery groups. The duration of the vasodilator effect of ATP was significantly greater than that of adenosine in both artery groups. CONCLUSION: These results suggest that ATP induces maximal dilation of coronary microvessels, most likely through an endothelium-independent mechanism. The degradation of ATP to adenosine 5'-monophosphate (AMP) and adenosine, as well as the direct action of ATP on A2-adenosine receptors may be responsible for the dilation. We conclude that coronary flow reserve can be determined safely with intracoronary ATP administration. PMID- 10361905 TI - Secondary prevention with lipid lowering therapy in familial hypercholesterolemia: a correlation between new evolution of stenotic lesion and achieved cholesterol levels after revascularization procedures. AB - OBJECT: To assess the value of secondary prevention with lipid lowering therapy following either balloon angioplasty (PTCA) or bypass surgery (CABG) in familial hypercholesterolemia patients, the correlation of the new evolution of stenotic lesions and therapeutically achieved cholesterol levels was studied in 50 patients. METHODS: All surviving patients were followed angiographically after 5 years, and findings were correlated with the annually determined total serum cholesterol (TC) levels. RESULTS: New coronary atherosclerotic plaques were not observed in 18 patients in whom the TC was controlled to <220 mg/dl but in 19 of 32 patients in whom the TC was >220 mg/dl, a new evolution of stenotic lesions was observed angiographically. CONCLUSION: The new evolution of stenotic lesions following revascularization in patients with FH can be controlled significantly by lipid lowering therapy to maintain a TC level of <220 mg/dl, and if diet alone can not achieve it, aggressive medication and even LDL apheresis might be justified. PMID- 10361906 TI - Association of human leukocyte antigen class II genes with autoantibody profiles, but not with disease susceptibility in Japanese patients with systemic sclerosis. AB - OBJECT: To examine the role of human leukocyte antigen (HLA) class II genes in the development of systemic sclerosis (SSc) as well as in the clinical and serologic expression of SSc in patients. METHODS: HLA-DRB1, DRB3, DRB4, DQB1, and DPB1 alleles were determined by genotyping; and serum antinuclear antibodies were identified using indirect immunofluorescence, double immunodiffusion and immunoprecipitation. PATIENTS: One hundred and five Japanese patients with SSc and 104 race-matched healthy controls. RESULTS: Frequencies of DRB1 and DQB1 alleles were not different between SSc patients and healthy controls, while DPB1*0901 was marginally increased in SSc patients. In contrast, SSc-related autoantibodies were closely associated with the clinical features. HLA class II genes were detected as follows: anti-DNA topoisomerase I antibody with diffuse cutaneous involvement, pulmonary fibrosis, and DRB1*1502-DQB1*0601-DPB1*0901; anti-U1RNP antibody with overlapping features of lupus and/or myositis and DRB1*0401/*0802-DQB1*0302; and anticentromere antibody with limited cutaneous involvement and DRB1*0101-DQB1*0501-DPB1*0402. In the analysis of the association of HLA class II and the clinical features in SSc patients significant differences were obtained only for the increased frequencies of arthritis and rheumatoid factor in patients with DRB1*0405 compared to those without. CONCLUSION: HLA class II genes strongly influence the production of SSc-related autoantibodies rather than the development of SSc. In addition, SSc is a composite disease of distinctive subsets defined by serum autoantibodies, which have specific clinical and HLA class II associations. PMID- 10361907 TI - Tuberous sclerosis associated with multiple hepatic lipomatous tumors and hemorrhagic renal angiomyolipoma. AB - We report a case of tuberous sclerosis associated with hepatic lipomatous tumors and renal angiomyolipomas. Abdominal ultrasonography revealed a high echoic large tumor in the left kidney. A provisional diagnosis of angiomyolipomas of the kidney was made based on computed tomography. Subsequent laparotomy revealed that the extracted tumor was renal angiomyolipoma. It was also revealed that there was an association with hepatic lipomatous tumors thought to be lipomas or angiomyolipomas by liver biopsy. Nearly half of all cases of angiomyolipoma in the kidney are reported as occasional association with tuberous sclerosis complex, but lipomatous tumors in the liver are rare. PMID- 10361908 TI - Malignant pheochromocytoma with multiple hepatic metastases treated by chemotherapy and transcatheter arterial embolization. AB - A 62-year-old Japanese male developed multiple hepatic metastases two years after resection of pheochromocytoma of the right adrenal gland. Transcatheter arterial embolization (TAE) was performed for the purpose of the treatment of hepatic metastases resistant to 27 cycles of combined chemotherapy consisting of cyclophosphamide, vincristine, and dacarbazine. After TAE, the hepatic metastatic lesions decreased in size and hypertension passed its crisis. The present case suggests the utility of TAE for multiple hepatic metastases under careful blood pressure monitoring. PMID- 10361909 TI - Localized pericarditis with calcifications mimicking a pericardial tumor. AB - A 62-year-old man was admitted with increasing palpitations. Radiography of the chest demonstrated a calcified mass. Magnetic resonance imaging revealed compression of the right ventricle by a tumor. At the time of cardiac catheterization, the coronary arteries were found not to supply blood flow of the mass, and no dip-and-plateau pattern was seen in the right ventricular pressure measurements. At the time of surgery, the mass was found to be a focal calcified thickening of the pericardium containing only pus. The thickening resembled an oval pericardial tumor. Microbiologic examination of the pus revealed Propionibacterium acnes. PMID- 10361910 TI - Idiopathic verapamil-sensitive left ventricular tachycardia complicated by right ventricular outflow tract ventricular tachycardia and ventricular fibrillation. AB - Idiopathic ventricular tachycardias (VTs) are generally divided into those arising from the right ventricle and those arising from the left ventricle. There has been few reports of two morphologically distinct VT occurring in patients with no apparent structural heart disease. We report a patient with verapamil sensitive left VT with a right bundle branch block pattern that spontaneously changed to VT with a left bundle branch block pattern. Ventricular fibrillation was induced by the application of programmed stimulation. Although it is unclear if our patient with pleomorphic VT has ventricular vulnerability, it is necessary to investigate further and follow him carefully. PMID- 10361911 TI - Prevention of premenstrual exacerbation of hereditary coproporphyria by gonadotropin-releasing hormone analogue. AB - A 20-year-old Japanese female needed frequent hospitalization due to premenstrual exacerbation of hereditary coproporphyria (HCP). Intranasal buserelin acetate, a gonadotropin-releasing hormone analogue, was given to suppress her menstrual cycles. Her porphyric symptoms subsided dramatically as she became amenorrhoeic. Urinary excretion of porphyrin derivatives fell significantly. She has been free from recurrent attacks, but suffers a minor porphyric attack once in 5 years. However, borderline osteopenia secondary to hypoestrogenism has been noted. Although these analogues are potent in suppressing estrogen-induced porphyric symptoms, due precautions should be taken to avoid bone demineralization in the long-term use. PMID- 10361912 TI - Bilateral pleuritis caused by Legionella micdadei. AB - A 58-year-old woman was hospitalized because of progressive respiratory distress. She had a history of myasthenia gravis and invasive thymoma. After thymectomy, she had been administered oral prednisolone and intrathoracic anti-cancer drugs postoperatively. Her chest radiograph revealed bilateral pleural effusions. Legionella micdadei (L. micdadei) was isolated from the pleural effusions, and she was diagnosed as pleuritis caused by L. micdadei. She died despite intensive therapy with mechanical ventilation, drainage tube in the chest and intravenous erythromycin. Although only two cases of Legionellosis caused by L. micdadei have been reported in Japan, clinicians should be aware of L. micdadei as one of the candidates for infection in immunosuppressed hosts. PMID- 10361913 TI - Intrabronchial Aspergillus nidulans infection in an immunocompetent man. AB - We describe the first report of intrabronchial Aspergillus nidulans infection in an immunocompetent patient, which fit the description of bronchocentric granulomatosis. The patient had a history of accidental aspiration of light grade oil. Fiberoptic bronchoscopy revealed that the right B4aii alpha was obstructed. Endobronchial biopsy specimens contained fungal hyphae. The fungus was confirmed to be Aspergillus nidulans by culture. We suspected that aspiration of light grade oil had injured the bronchial mucosa, after which airborne Aspergillus nidulans had entered the lesion and multiplied. Intrabronchial fungal infection can occur in a healthy person without immunologic abnormalities, if a bronchial lesion provides an entry portal. PMID- 10361914 TI - Successful treatment of adult-onset Henoch-Schonlein purpura nephritis with high dose immunoglobulins. AB - A 26-year-old woman was admitted for the evaluation of edema and massive proteinuria. She had a history of purpura of the lower extremities, abdominal pain and melena. Laboratory investigations showed hypoalbuminemia, hypercholesterolemia and proteinuria of over 10 g/day. Renal biopsy showed moderate proliferative glomerulonephritis with mesangial immunoglobulin A (IgA) deposition. She was diagnosed as having Henoch-Schonlein purpura nephritis. Oral prednisolone, dipyridamole and intravenous heparin treatment were not effective. Steroid pulse therapy induced a partial improvement of proteinuria to 2-3 g/day. High-dose intravenous immunoglobulin (i.v.-IG) treatment was introduced and a dramatic improvement of proteinuria was noted. I.v.-IG should be fully considered in patients with steroid-resistant Henoch-Schonlein purpura nephritis. PMID- 10361915 TI - A care package for managing female sexual assault in genitourinary medicine. AB - This paper describes the development of a designated in-house service for the management of adult female victims of sexual assault within the Department of Genitourinary Medicine (GUM) at St Mary's Hospital, London. This was set up in 1994 as a need was identified by medical, nursing, psychological and health advising staff for an appropriate streamlined service which would provide comprehensive sexual health screening, psychological support and therapy and adequate medico-legal documentation within the limitations of a busy GUM clinic. A structured package of care consisting of medical and psychological protocols with training for relevant staff and a specialist in-house referral clinic was introduced. Fifty-four patients were seen during the first 17 months of the service, the notes of 48 of these were examined and relevant epidemiological and audit data are presented here. By auditing the quality of documentation before and after the introduction of the protocols specifically looking at the appropriateness and comprehensiveness of the sexually transmitted diseases screen and the medico-legal documentation it was clear that the quality of care to these patients was improved. We present here the development of these protocols, a detailed description of the protocols themselves and the method of their implementation. PMID- 10361916 TI - Neisseria gonorrhoeae in Newcastle upon Tyne 1995-1997: increase in ciprofloxacin resistance. AB - Fluoroquinolones and third generation cephalosporins are the most effective antimicrobial agents for the treatment of gonorrhoea. However, clinically significant resistance to fluoroquinolones in Neisseria gonorrhoeae has been reported worldwide including Britain. The aim of this analysis was to study the factors relating to ciprofloxacin resistance and treatment failure. A total of 201 patients attending the Newcastle Genitourinary Medicine (GUM) clinic from 1995-1997 who were diagnosed with culture positive gonorrhoea was analysed. Treatment failure rates for ciprofloxacin were determined and the minimum inhibitory concentration (MIC) was measured for all cases of treatment failure. The case notes of all patients who had strains with MICs of ciprofloxacin in the resistant range (>0.05 microg/ml) were reviewed to determine the clinical outcome. The ciprofloxacin resistance with treatment failure was seen in 5% (8/160). All the 8 cases of treatment failure were heterosexual and had isolates resistant to penicillin and 4 cases (50%) were also resistant to tetracycline. All were sensitive to spectinomycin and ceftriaxone. Most of the cases probably acquired their infection from the Far East. As ciprofloxacin resistance seems to be associated with overseas exposure, changes in the standard treatment of gonorrhoea are not justified but consideration should be given to appropriate alternatives when the infection may have arisen from where such resistant strains are endemic. Monitoring fluoroquinolone resistance is now essential for ensuring adequate treatment of infections with resistant strains and for maximizing the time of usage of fluoroquinolones to treat gonorrhoea. PMID- 10361917 TI - Trends in HIV-related consultation in Dutch general practice. AB - New medicine against AIDS and a possible changing attitude towards AIDS will affect the role of the general practitioner (GP). We aim to explore and assess the changing role of the GP in preventing and detecting AIDS, which will be done by providing insight into the changing numbers and content of HIV-related consultations in the general practice. Since 1988 a representative sample of 63 Dutch GPs have participated in a network. They recorded all face-to-face consultations with non HIV-infected patients in which the subject of AIDS was brought up. Timetrend analysis is used to investigate variations over time in the number and content of the consultations, GPs' actions and patients' characteristics. The influence of a rural or urban setting and the characteristics of the patients who are involved are also taken into account. Until 1994 a significant increase was found in the number of consultations. In highly urban areas the number of consultations is higher and still growing, whereas physicians in rural areas see fewer patients every year. The most important topic of conversation was the request for an HIV test (74%). This figure grew over the years, as did the number of tests performed. GPs became less passive and restrictive in advising tests. The group of patients has also changed, e.g. patients do not mostly belong to traditional risk groups anymore, and are significantly younger. AIDS seems to have become more familiar to patients and doctors. A lot of general information is available from different sources. Because of this change in attitude and knowledge of patients, the GPs' role as it relates to AIDS is becoming more specific in tracing infected patients and giving customized information to individuals. Patients visit their physicians less often because of concerns about AIDS, but the GP continues to fulfil a very important role in the prevention and detection of AIDS. PMID- 10361918 TI - Seroprevalence of HIV, HBV, and syphilis and associated risk behaviours in male transvestites (Hijras) in Karachi, Pakistan. AB - In Pakistan, male sex workers are predominantly transvestites and transsexuals known as Hijras. In 1998 in Karachi, Pakistan, we studied the seroprevalence of HIV, HBsAg and syphilis and associated risk factors in Hijras. Study subjects were verbally administered a structured questionnaire that determined risk behaviours for sexually transmitted and blood-borne diseases and knowledge of AIDS/STDs. After pre-test counselling, verbal consent was taken for serological testing. The results were provided on a one-on-one post-test counselling session. Three hundred male transvestites were approached; all agreed to answer the questionnaire, 208 consented to blood testing. Of 300, 81% acknowledged commercial sex with men. Of 208, prevalence of syphilis was 37%; HBsAg 3.4%; HIV 0%. The prevalence of HIV and hepatitis B virus (HBV) is low in transvestite sex workers but that of syphilis is high. Intervention programmes implemented at this stage can have an impact on HIV and STD prevention. PMID- 10361919 TI - Bacterial vaginosis in lesbians: evidence for lack of sexual transmission. AB - The effect of non-heterosexual factors on the vaginal flora has been studied. Ninety-one lesbians attending a specialist genitourinary medicine service for lesbians were studied. Bacterial vaginosis (BV) was diagnosed in 51.6% of them. While most of the women had previously had a male sexual partner, the presence of BV was not associated with a male sexual partner in the previous 12 months. A detailed analysis of lesbian sexual practices in the group did not relate BV to any sexual practice which would have the propensity to pass vaginal secretions from one to the other. PMID- 10361920 TI - Treatment delay and reliance on private physicians among patients with sexually transmitted diseases in China. AB - We examined health-care seeking practices among patients with sexually transmitted diseases (STDs) in south China. In 1995, we recruited a consecutive sample of 939 STD patients attending the STD clinics of the Municipal STD Control Centers of Guangzhou and Shenzhen, 'special economic zones' near Hong Kong. Attending physicians interviewed patients face-to-face using a standard survey questionnaire. Twenty-seven per cent of all subjects had sought treatment elsewhere for their presenting complaints, before visiting a study clinic. The main sources of prior treatment were private physicians followed by public clinics and drugstores. Women were more likely than men to delay in presenting their current symptoms to a study clinic (32% vs 25%, P=0.046). Factors associated with treatment delay differed by gender. Among men, seeking prior treatment from private physicians (OR=3.31; 95% CI=1.70, 6.43), having no urethral discharge (OR=4.00; 95% CI=2.33, 6.85), having engaged in sex trade (OR=1.64; 95% CI=1.03, 2.63), or being a resident in Shenzhen (OR=1.80; 95% CI=1.12, 2.89) were more likely to delay seeking treatment. Among women, only living in Shenzhen (OR=2.86; 95% CI=1.56, 5.25) was associated with treatment delay. Promotion of appropriate health-seeking behaviours and better management of STDs must be a top priority to slow a rapid spread of STD/HIV in China. Health education, improvement of STD care in the public and private sectors, and regulations of unauthorized private physicians, may help with STD control and HIV prevention. PMID- 10361921 TI - An epidemiological evaluation of the use of microbiological tools for identifying gonorrhoea infection networks. AB - We aimed to assess the utility of various techniques for identifying gonorrhoea infection networks. All residents of a non-metropolitan North Carolina county visiting a sexually transmitted disease (STD) clinic during a 17-month period were screened for gonorrhoea. Infection networks were estimated by serovar type combined with antibiotic resistance, arbitrarily primed polymerase chain reaction (AP-PCR), or temporal clustering. The residential addresses of infected patients were geocoded and mapped. Among 2 serovar types, the presence of distinguishing characteristics of a network, based on questionnaire data, was assessed with prevalence ratios and 95% confidence intervals (CIs) relative to those not in the network. Twenty-five serovar types were identified among 759 gonorrhoea infections. In one serovar, the networks further delineated by temporal clusters correlated with particular AP-PCR types. In most instances, however, different typing techniques painted different network pictures. No refined serovar network stood out as having a particular set of characteristics that could be used to shape intervention. Teasing out an individual infection network with unique characteristics will require the development and use of other microbiological tools. PMID- 10361922 TI - Inadequate treatment for sexually transmitted diseases in the South African private health sector. AB - Correct management of sexually transmitted diseases (STDs) is important for their control, and to reduce HIV transmission. Guidelines on syndromic management of STDs were introduced by the provincial Department of Health in KwaZulu/Natal (KZN) in South Africa in 1995. The drug treatment provided for STDs by the 11 private general practitioners in one rural district was assessed and compared with provincial guidelines. Information was gathered through semistructured interviews which asked the 11 doctors, who all dispense prescribed drugs as part of the consultation fee, how they would treat 3 hypothetical cases of STD syndromes. In all 33 prescriptions, the treatment did not correspond exactly with provincial recommendations and only 3 (9%) were adequate. All other prescriptions were inadequate because dose or duration was incorrect in 6 (18%), or because incorrect drugs were prescribed in 24 (73%) of cases. Eight of the 11 doctors did not provide adequate treatment for any of their cases. A continuing medical education programme for the doctors and their staff was devised to improve the STD treatment in the private sector in this South African district. PMID- 10361923 TI - Routine reporting or sentinel surveys for HIV/AIDS surveillance in resource-poor settings: experience in South Africa, 1991-97. AB - Information from routine and sentinel surveillance was used to monitor the HIV/AIDS epidemic in KwaZulu-Natal, South Africa between 1991 and 1997. Comparisons were made between data obtained from (1) sentinel surveillance for antenatal HIV infection, pulmonary tuberculosis (PTB), and AIDS in a single health district and (2) province-wide sentinel surveillance for antenatal HIV infection, legally required notification of cases of PTB, and voluntary notification of AIDS cases. HIV prevalence among antenatal clinic attenders in the sentinel district rose rapidly and at similar rates to provincial figures: 4.2% vs 4.8% in 1992 to 25.9% vs 26.9% in 1997. PTB incidence increased four-fold in the sentinel district over the study period, whereas provincial PTB figures from passive surveillance fluctuated widely and showed no clear increase (Chi square for trend 425.5, P<0.00001). AIDS incidence in the sentinel district increased dramatically while provincial data from the voluntary reporting system showed a less consistent and much slower rise (Chi-square for trend 9.07, P=0.003). Incidence of AIDS in 1997 was estimated as 437/10(5) in the sentinel district compared to 32/10(5) in the provincial figures. Routine disease notification and voluntary reporting systems are likely to underestimate the impact of the HIV/AIDS epidemic in resource-poor settings. Sentinel surveillance at representative sites should be developed to validate or replace passive surveillance systems. PMID- 10361925 TI - A case of cat scratch disease masquerading as lymphogranuloma venereum. PMID- 10361924 TI - Chlamydia trachomatis infection in women with and without cervical intraepithelial neoplasia. AB - The aim of the study was to compare the frequency of Chlamydia trachomatis infection in patients with cervical intraepithelial neoplasia (CIN) and in women without cervical pathology. In a study group of 423 patients with histologically proven CIN and in 108 controls with normal cervical smear, cytological material for direct immunofluorescence analysis was obtained. Among 423 patients, 24 (5.7%) had CIN 1, 108 (25.5%) CIN 2 and 291 (68.8%) CIN 3. Among all patients with CIN, 27 (6.4%) were C. trachomatis positive and 396 (93.6%) C. trachomatis negative. In the control group 6 (5.6%) were C. trachomatis positive and 102 (94.4%) C. trachomatis negative. The difference between C. trachomatis infection incidence in patients with CIN and in women without cervical pathology was not significant (chi2=0.29; P>0.05). In this study, no difference in C. trachomatis infection incidence was detected between patients with CIN and women with normal cervical smears. The impact of C. trachomatis infection seems not to interfere with the development or even the promotion of CIN. PMID- 10361926 TI - Simultaneous occurrence of HTLV-I associated myelopathy, uveitis and smouldering adult T cell leukaemia. GIPH (Interdisciplinary HTLV-I/II Research Group). PMID- 10361927 TI - HIV/AIDS in Bangladesh: an assessment of biomedical risk factors for transmission. AB - Behavioural risk factors for HIV/AIDS in Bangladesh were reviewed in a preceding article in this journal. Omitted from that review was a discussion of potential biomedical risk factors including: (i) an unregulated blood supply system in which blood used in transfusions is not screened for HIV and is donated primarily by professional donors: (ii) unsterile injections in non-formal and formal health care settings; and (iii) a high prevalence in high-risk groups of other sexually transmitted diseases (STDs) which may function as co-factors for HIV transmission, particularly if chronically untreated. Studies elsewhere in the world suggest that the unregulated blood supply system, in particular, poses a serious danger in terms of the spread of the HIV epidemic. While certain socio cultural factors may be contributing to low levels of HIV in Bangladesh, the prevalence of biomedical and behavioural risk factors suggest the importance of implementing targeted cost-effective interventions now. PMID- 10361928 TI - Prevention of pelvic infection: room for improvement. PMID- 10361929 TI - Outcome of inflammatory smears managed in a genitourinary medicine clinic. PMID- 10361930 TI - Autonomic regulation of lymphatic flow in the lower extremity demonstrated on lymphoscintigraphy in patients with reflex sympathetic dystrophy. AB - PURPOSE: Nuclear medicine techniques were used to show that the peripheral lymphatics are under autonomic control in much the same way as the blood vessels that supply the same anatomic region. METHODS: Three patients with complex regional pain syndrome type 1 (reflex sympathetic dystrophy) involving a lower extremity were evaluated using three-phase bone scintigraphy and peripheral lymphoscintigraphy. Each patient was treated with ipsilateral chemical lumbar sympathectomy, and lymphoscintigraphy was repeated within several days of the procedure. RESULTS: All three patients had evidence of decreased flow (compared with the contralateral extremity) to normal flow after ipsilateral sympathectomy. Bone scintigraphy, before and after sympathectomy, was difficult to interpret because of the effects of altered weight bearing. Two patients who had unilateral peripheral edema showed marked improvement after sympathectomy and increased lymphatic flow. CONCLUSIONS: Peripheral lymphatic function is controlled by the autonomic nervous system. In reflex sympathetic dystrophy, peripheral edema may be caused by an increased sympathetic stimulus to the lymphatics. Further study of this phenomenon may show that nuclear medicine studies, such as bone scintigraphy and lymphoscintigraphy, can be used to distinguish patients who will benefit from sympathectomy from those who will not, thereby obviating invasive testing and unnecessary invasive treatment. PMID- 10361931 TI - Direct lymphatic drainage from a melanoma on the back to paravertebral lymph nodes in the thorax. AB - Preoperative lymphoscintigraphy with Tc-99m antimony sulfide colloid was performed in a patient with cutaneous melanoma on the lower back just to the right of the midline. There was direct lymphatic drainage to paravertebral nodes in the chest on the right side at the level of the sixth and seventh thoracic vertebrae. There was also drainage directly to the right groin and via a series of interval nodes to the right axilla. Knowledge of the presence of such drainage may influence the surgical management of patients. PMID- 10361932 TI - Sentinel node detection and definition may depend on the imaging agent and timing. AB - PURPOSE: Two cases of sentinel lymph node imaging are presented in which the results are exceptions to what the literature generally defines as sentinel lymph nodes. In one case, Tc-99m antimony trisulfide colloid produced significantly different results than did Tc-99m tin colloid. In the second case, the results bring into question the definition of a sentinel node as the first in a lymphatic drainage pathway. MATERIALS AND METHODS: In one patient, lymphoscintigraphy was performed initially using Tc-99m antimony trisulfide colloid injected intradermally around a melanoma excision site. Repeated lymphoscintigraphy 1 month later, 1 hour before sentinel node excision, was done using Tc-99m tin colloid, a larger particle than antimony trisulfide colloid. The second patient, with a melanoma biopsied only, had sentinel node imaging performed using Tc-99m sulfide colloid, a particulate also larger than antimony trisulfide colloid and also 1 hour before sentinel node excision. RESULTS: In the first patient, imaging with the smaller antimony trisulfide colloid showed more lymphatic pathways and more sentinel nodes than with tin colloid. In the second patient, the first focus of retention of the imaging agent in the lymphatic pathway seen showed less intense accumulation than the next focus in the pathway, contrary to published reports that the sentinel node shows more intense accumulation than do nodes further downstream in a lymphatic pathway. CONCLUSIONS: There are exceptions to published characteristics of sentinel node lymphoscintigraphy, so care must be exercised in localizing sentinel nodes. PMID- 10361933 TI - Bone SPECT of the back after lumbar surgery. AB - PURPOSE: It may be difficult to evaluate back pain in patients who have undergone spinal surgery, because symptoms may be secondary to all the possible abnormalities in patients who have not had surgery plus postoperative complications, including infection, unstable fusion sites, or transfer of biomechanical stresses to other regions. MATERIALS AND METHODS: Sixty-three patients with back pain and a history of lumbar spinal surgery had bone SPECT examinations. Twenty-eight patients had laminectomies, 10 had laminectomies with fusion, 10 had laminectomies with fusion and metallic stabilization devices (3 of which were removed), 7 had fusion without laminectomy, 7 had discectomies, and 1 had a fusion with metallic stabilization but no laminectomy. Eighty-seven percent of the fusions were posterior. The results of SPECT scanning were correlated with surgery, clinical information, and diagnostic radiologic studies. RESULTS: Patients with fusions tended to be scanned further out from the time of surgery than were patients with laminectomy alone or especially discectomy. Bone SPECT excluded bony abnormalities in the operative site in 7 of 63 patients. One hundred thirty-two lesions were uncovered, with facet abnormalities (n = 51) the most common followed by disc space-centered conditions (n = 29), pseudarthrosis (n = 20), sacroiliac joint (n = 18), vertebral body lesions (n = 9), and miscellaneous sites (n = 5). Sixty percent of the abnormalities located in the facets, disc spaces, and vertebral bodies were located in the operative field, whereas 29% were above and 11% were below it. CONCLUSION: Bone SPECT was useful in evaluating these patients to exclude bony lesions or to identify pseudarthrosis, abnormal facets, disc space-centered lesions, and sacroilitis. PMID- 10361934 TI - Anterior chest wall pain in postpartum costochondritis. AB - Costochondritis is a common diagnosis in patients with anterior chest wall pain in whom serious disease has been excluded. The diagnosis is usually made on clinical grounds, because laboratory and imaging investigations usually provide little information. The authors describe a young woman with postpartum costochondritis and discuss the role of bone scintigraphy in confirming the clinical diagnosis. PMID- 10361935 TI - The benefit of SPECT when added to planar scintigraphy in patients with bone metastases in the spine. AB - PURPOSE: This study compared the efficiency of SPECT with planar bone scans in differentiating malignant from benign lesions and in detecting metastases to the spine. METHODS: Planar scintigraphy and SPECT were performed in 37 patients with low back pain without known malignancy and in 38 patients with confirmed malignancy. The type, location, and intensity of tracer accumulation were compared on the planar and SPECT scans. The malignant or benign nature of lesions was proved by radiologic methods, histologic findings, 6 month follow-up, or all of these. RESULTS: More metastases were detected by SPECT (SPECT, 58 of 64; planar, 42 of 64; P < 0.01). In three of seven patients with known malignancy who had a normal result of planar scan, only SPECT detected metastases. Fifty-nine metastases were radiologically mainly osteolytic, one was osteoblastic and four were mixed. Most lesions showed increased radioactivity (40 of 42 on planar scans vs. 45 of 58 on SPECT) and 2 of 42 (5%) vs. 12 of 58 (21%) were cold with marginally increased uptake. One of 58 metastases was a cold lesion seen on SPECT only. Lesions were more often malignant than benign when seen on SPECT in a pedicle (n = 5; malignant = 3, benign = 2), in the body and pedicle (n = 22; malignant = 14, benign = 8), within the vertebral body (n = 5; malignant = 4, benign = 1) and in the whole vertebra (n = 6; malignant = 4, benign = 2). The lesion to background ratio was higher on SPECT than on planar scans (SPECT, 2.26; planar scans, 1.86; P < 0.05 in malignant lesions). CONCLUSIONS: SPECT of the spine improved the diagnostic accuracy of bone scans when added to a planar scan in patients with known malignancy and clinical suspicion of spinal metastases when the planar scan was borderline abnormal. It helps in differentiating between benign and malignant lesions of the spine. PMID- 10361936 TI - Resolved splenic accumulation of Tc-99m HMDP after recovery of disseminated intravascular coagulation in a patient with rhabdomyosarcoma. AB - Unusual, intense splenic radioactivity was seen on bone scintigraphy with Tc-99m HMDP in a 14-year-old boy with alveolar rhabdomyosarcoma complicated by disseminated intravascular coagulation. Abnormal splenic radioactivity was resolved after recovery from the disseminated intravascular coagulation. During treatment of disseminated intravascular coagulation and tumors, the patient received repeated blood transfusions, resulting in iron overload, but this did not prevent the abnormal splenic uptake from resolving. This case indicates that disseminated intravascular coagulation may be a cause of splenic accumulation of bone-seeking agents, and that abnormal splenic uptake can be resolved. PMID- 10361937 TI - I-123 MIBG cardiac imaging in diabetic neuropathy before and after epalrestat therapy. AB - I-123 metaiodobenzylguanidine (MIBG) scintigraphy is a new method to evaluate cardiac sympathetic nerve disturbance in patients with diabetes mellitus. Epalrestat specifically inhibits aldose reductase and improves diabetic neuropathy. The authors report a case of improvement in cardiac sympathetic dysfunction using MIBG scintigraphy with epalrestat therapy. In this case, epalrestat effectively reversed diabetic neuropathy, and MIBG scintigraphy was useful to evaluate the effect of epalrestat. PMID- 10361938 TI - Familial occurrence of congenital hypothyroidism due to lingual thyroid gland. AB - Two sisters who presented with midline neck masses proved to be biochemically hypothyroid. Thyroid scintigraphy supplemented with perchlorate discharge testing showed lingual thyroid glands and ruled out the expected autosomal recessive organification defect. The related literature is reviewed. PMID- 10361939 TI - Tc-99m dextran: a new and sensitive general purpose scintigraphic agent for diagnosing intestinal inflammation. AB - PURPOSE: This feasibility study was undertaken to compare the sensitivity of Tc 99m dextran with that of Tc-99m human immunoglobulin G to diagnose ulcerative colitis, and to explore its possible role in disease follow-up. MATERIALS AND METHODS: Twenty-six patients with active disease and six patients in remission underwent serial Tc-99m dextran scanning for as long as 3 hours or more after injection. Eight of the patients with active disease also underwent Tc-99m human immunoglobulin G imaging. RESULTS: Twenty-four of 26 (92%) patients with active ulcerative colitis had a positive result of the Tc-99m dextran study, mainly within 1 hour. In comparison, Tc-99m human immunoglobulin G accumulated abnormally in four of eight (50%) patients and had a relatively poor target localization with high persisting background even after 6 hours. Four of the six patients in remission still had a positive result of the dextran scan, but the abnormal uptake was less than that in the patients with active disease. The disease has recurred already in one of these patients. A patient with pancolitis who was receiving steroid enema therapy had intense uptake of Tc-99m dextran in the ascending colon, probability because it was outside the range of the enema. CONCLUSIONS: Tc-99m dextran is a sensitive and cost-effective agent to diagnose ulcerative colitis, and it may have a role in disease follow-up. PMID- 10361940 TI - Use of Tc-99m DTPA galactosyl human serum albumin to predict postoperative residual liver function. AB - PURPOSE: Residual liver uptake at 15 minutes (RLU15), a new index for predicting residual liver function after excision of hepatocellular carcinoma, was evaluated using dynamic liver images and SPECT by Tc-99m DTPA galactosyl human serum albumin. MATERIALS AND METHODS: In 13 cases of hepatectomy, RLU15 was compared with postoperative serum prothrombin levels, serum bilirubin levels, and ascites. RESULTS: RLU15 showed good correlation with both the prothrombin activity and the serum bilirubin, with correlation coefficients of r = 0.829 and r = -0.757. CONCLUSION: This new index appears useful for predicting residual liver function after hepatectomy. PMID- 10361941 TI - Tc-99m DTPA used as reference imaging to evaluate the distribution of other tumor seeking tracers in tumors associated with neurofibromatosis. AB - Two patients with multiple benign and malignant tumors associated with neurofibromatosis underwent radionuclide imaging with Tc-99m DTPA, Tl-201, and Ga 67. In these patients, Tc-99m DTPA accumulated intensively in both the benign and malignant tumors and localized and defined the extent of every tumor. In contrast, Ga-67 and Tl-201 uptake was seen only in focal areas of tumor where there was malignant transformation or at sites that showed progressive tumor growth. Tc-99m DTPA imaging accurately demonstrated areas of neoplastic involvement and identified the areas that would be seen with the other two tracers in individual tumors. Tc-99m DTPA may not always be used for the differential diagnosis of malignant and benign tumors of neurofibromatosis, but it can provide a reference pattern for imaging to evaluate accurately the distribution of Tl-201 and Ga-67 by mapping out the anatomic extent of these tumors. PMID- 10361942 TI - Ureteral leak after renal transplantation. AB - In the course of radionuclide dynamic studies in 187 consecutive patients who had renal transplants, leakage from a ureter was identified in five cases (an incidence of less than 3%). Demonstrated patterns fell into three classes: 1) perirenal collection (n = 2), 2) peribladder activity (n = 2), and 3) downward dissection (n = 1 case). The sensitivity and specificity of the radionuclide study for detecting ureteral escape of urine after renal transplantation is unknown. However, this study suggests that both indices may be high. PMID- 10361943 TI - Asymptomatic fatigue fracture of the proximal ulna associated with symptomatic fatigue fractures of the lower limbs. PMID- 10361944 TI - Symmetric avascular necrosis of the shoulders and hips in sickle cell disease. PMID- 10361945 TI - Bone scintigraphy in polyostotic fibrous dysplasia. PMID- 10361946 TI - Transient right-to-left shunt in acute pulmonary embolism. PMID- 10361947 TI - Ga-67 citrate uptake in bilateral endogenous endophthalmitis. PMID- 10361948 TI - Focal nodular hyperplasia of the liver: scintigraphic demonstration using three hepatic imaging agents. PMID- 10361949 TI - Tc-99m MIBI uptake by a male breast lymphoma accompanied by diffuse bone marrow metastases. PMID- 10361950 TI - Incidental finding of hiatal hernia uptake on Tc-99m sestamibi breast imaging. PMID- 10361951 TI - Detection of tubular intestinal duplication by a Meckel's scan. PMID- 10361952 TI - Tc-99m phytate images in a patient with a wandering spleen. PMID- 10361953 TI - In-111 pentetreotide scintigraphy to monitor nonsecreting neuroendocrine tumors. PMID- 10361954 TI - Intrathoracic goiter. PMID- 10361955 TI - Testicular I-131 6beta-iodomethyl-19-norcholesterol uptake in a patient with adrenogenital syndrome. PMID- 10361956 TI - Avid Ga-67 uptake in active celiac disease. PMID- 10361957 TI - Tc-99m sestamibi uptake in a patient with gynecomastia: a potential pitfall in the diagnosis of breast cancer. PMID- 10361958 TI - Current readings in nuclear medicine. PMID- 10361959 TI - Social phobia: diagnosis, severity and implications for treatment. AB - Social phobia has a direct effect on the ability of the individual to interact with others in social or work situations and as a result is associated with a high level of dysfunction. The level of impairment is as severe as that found in other chronic disorders such as depression and is increased by a cascade of comorbidity, which complicates management. Efficacy in social phobia was reported with the MAOIs and this led to the investigation of reversible inhibitors of monoamine oxidase-A (RIMA) which offer a safer alternative. The efficacy of moclobemide has been shown in social phobia and the effect is more clear cut in patients with more severe symptoms. The better effect in severe social phobia has also been reported with the SSRI paroxetine. Effective treatment of social phobia appears to be able to overcome the substantial chronicity of illness which is frequent in sufferers. The introduction of these treatments should encourage people with social phobia to seek medical help for this hitherto much neglected disorder. PMID- 10361960 TI - Moclobemide and fluoxetine for panic disorder. International Panic Disorder Study Group. AB - An international, multicentre, double blind parallel group study compared the tolerability and efficacy of moclobemide with the selective serotonin reuptake inhibitor (SSRI) fluoxetine for panic disorder. SSRIs have been shown effective for panic. The target dose of moclobemide was 450 mg and of fluoxetine was 20 mg. There were two consecutive studies. An eight week study of acute adverse events, tolerability and efficacy was followed by a long-term extension study to 1 year. The efficacy data showed no significant difference between moclobemide and fluoxetine. Both had acute efficacy, with 63% moclobemide and 70% fluoxetine patients (ns) panic free at 8 weeks. Both agents were well tolerated to 8 weeks, but moclobemide had fewer severe adverse events (5) than fluoxetine (9). There were no severe adverse events in the extension phase with either drug, and almost all patients completing 1 year extension treatment (moclobemide 61 patients, fluoxetine 65) were much or very much improved. These data suggest moclobemide and fluoxetine are tolerated and effective for both acute panic treatment and maintenance therapy. PMID- 10361961 TI - The neurobiology of social phobia. AB - Although Social Phobia has been recognised for centuries in comparison with other anxiety disorders, relatively little work has been done to understand its neural basis. The present review attempts to redress this balance by giving an overview of the current state of knowledge in this disorder. By putting together data from the treatment responses to specific agents, the effects of chemical challenges which have been used in other anxiety disorders and by reviewing data on central and peripheral neurotransmitter and endochrine abnormalities, it is possible to begin to generate some potentially testable theories of aetiology and mechanisms. Finally, we review the potential use of neuroimaging techniques to better detail the brain circuits and possibly neurotransmitters involved in social phobia, showing some of our preliminary work using (15)O water blood flow PET activation studies to determine the brain circuits in which metabolism is changed during the experience of social anxiety. PMID- 10361963 TI - Posterior fossa tumors in children: indications for ventricular drainage and for VP shunting. PMID- 10361962 TI - The efficacy and safety of moclobemide compared to clomipramine in the treatment of panic disorder. AB - The primary objectives of this multicenter study were to determine the efficacy and safety of moclobemide, a selective reversible inhibitor of monoamino oxidase A, as drug treatment in DSM-III-R panic disorder with and without agoraphobia. In a comparative double-blind, randomized parallel-group design with fixed-flexible dose moclobemide 450 mg per day was compared to clomipramine 150 mg per day, as that drug was considered standard treatment of panic disorder in Europe. 135 patients were randomized and treated for a period of eight weeks. No other treatment was given. By the end of week 8, 49% of the patients treated with moclobemide and 53% of those treated with clomipramine were seen as treatment responders since they were without panic attacks. 78% of the patients in the moclobemide and 88% in the clomipramine group were considered responders according to clinical global impression of change. No significant differences were found between the two treatments at week 8. Adverse events were observed with significantly higher frequency among patients treated with clomipramine, particularly due to anticholinergic side effects. Close to 20% of those treated with moclobemide experienced headache, dizziness, nausea, insomnia, or dry mouth, but other adverse effects were infrequent. In conclusion, moclobemide in a dose of 450 mg per day seems to be a good drug alternative for treatment of panic disorder with and without agoraphobia. PMID- 10361964 TI - Presurgical strategies and epilepsy surgery in children: comparison of literature and personal experiences. AB - We reviewed 41 studies on epilepsy surgery in 1645 children and adolescents. The available data vary greatly from one paper to another, particularly as regards the age of the patients included, selection criteria, presurgical methodology, and the type of surgical intervention. Moreover, the surgical results are classified according to very different criteria. Our experience in Grenoble concerning 55 children (age < or = 16 years; postoperative follow-up > 18 months) is discussed in the light of data in the literature. The need is stressed for a consensus on the type of information that must be supplied by well-detailed studies on homogeneous groups of epilepsy surgery patients. PMID- 10361965 TI - Hydrocephalus and epileptic seizures. AB - Is there an association between shunted hydrocephalus and the development of epileptic seizures? To answer this question a retrospective review of the medical records of 197 patients with shunted hydrocephalus was undertaken. In this series 17% of patients with hydrocephalus developed seizures. No correlation was found between the occurrence of epileptic seizures and a shunt malfunction, the number of shunts placed, the age of the patient at the initial shunt procedure or the location of the shunt. Patients with hydrocephalus who had significant cognitive delay or significant motor disability were significantly more likely to develop seizures than patients who did not. The findings of this review support the hypothesis that the occurrence of seizures in children with hydrocephalus is related to an underlying diffuse encephalopathy and not to the hydrocephalus or to procedures related to the treatment of this disorder. PMID- 10361966 TI - Congenital torticollis in association with craniosynostosis. AB - The incidence of congenital torticollis in association with plagiocephaly is 1 in 300 newborns, with the torticollis resulting from pathologically sustained contraction of the sternocleidomastoid. Such conditions as facial asymmetries, craniovertebral anomalies, cervical hemivertebra, and mono- or polydysostoses may also be associated with torticollis diagnosed during the neonatal period. With particular reference to synostotic (coronal and/or lambdoidal) plagiocephaly, a clear distinction is made in this paper between posterior neurocranial flattening secondary to the sustained rotation of the skull resulting from torticollis and that seen in synostotic plagiocephaly. The rarity of torticollis with sustained contraction of the sternocleidomastoid muscle relative to the frequency of occipital-parietal flattening in newborn kept in the supine position has not been discussed in the literature and is therefore of clinical importance. In light of the fact that the prognosis and, consequently, the treatment plan vary directly with the presence or absence of synostoses, clinical evaluation also includes cephalometrics, plain skull X-rays, and CT imaging. If the torticollis is associated with neurocranial deformity but synostosis is absent, cervical traction and physiotherapy resolve the symptoms. When, however, the clinical picture is complicated by synostotic plagiocephaly, corrective surgery is necessary, though cervical traction and physiotherapy are essential to provide early and complete cure of the torticollis. PMID- 10361967 TI - Epidermoid cysts of the pineal region. AB - Localization of epidermoid cysts to the pineal region is rare. The 7-year-old boy now reported presented with an 18-month history of progressive ataxia. CT and MRI scans demonstrated a 2.5x2.5 cm cyst at the pineal region with obstructive hydrocephalus. At surgery via an occipital transtentorial approach, a characteristic "pearly tumour" was encountered, and complete resection was achieved. We present the management of this child with pineal region epidermoid cyst and review 11 cases reported in the literature since 1968. In all, 8 of the 12 patients were males. The age at the time of diagnosis ranged from 7 years to 69 years. Parinaud's syndrome and hydrocephalus are the most common presenting findings. All but 1 patient underwent direct surgical resection; 1 had stereotactic decompression. Surgical treatment brought about complete resolution of the presenting symptoms and signs in 10 of the 12 cases. One patient had persistent upgaze palsy. One patient died from progression of the pineal region mass. This patient presented with hemiparesis, which is a marker of clinical aggressiveness. The authors advocate direct surgical attack as opposed to stereotactic diagnosis and aspiration to: (1) obtain maximal resection and thereby limit the potential for recurrence and delayed complications of the cyst; (2) possibly avoid shunt placement in patients who present with hydrocephalus; and (3) decrease the likelihood of sampling error. PMID- 10361968 TI - A simple method of predicting hormonal outcome in children with intracranial germinoma. AB - Even a small intracranial germinoma (GE) frequently provokes pituitary hypofunction. We evaluated the relationships between preoperative hormonal status and hormonal outcome and between size of suprasellar mass and hormonal outcome in 22 children with intracranial pure GE to determine whether or not these can reflect the degree of hypothalamo-pituitary axis destruction. Preoperative hormonal status was graded from I to IV according to the serum prolactin level (s PRL) and thyroid function (TF). The hormonal outcome was estimated by the type and the number of hormonal replacement medications taken on the basis of the triple load test and endocrinological examinations at the time of the last follow up (median period: 43 months). Fifteen of the 22 patients had suprasellar lesions. All 13 patients who had diabetes insipidus (DI) at presentation needed desmopressin acetate (DDAVP) during the follow-up period. The correlation between increment of preoperative grade and the increment in the number of hormones to be replaced was statistically significant (P<0.05). This preoperative grading was a more reliable predictor than the size of suprasellar tumor. In conclusion, preoperative grading by s-PRL and TF test is useful for the prediction of posttreatment hormonal replacement in children with intracranial pure GE. PMID- 10361969 TI - Treatment of intracranial nongerminomatous malignant germ cell tumor in children: the role of each treatment modality. AB - To investigate the role of surgical tumor resection, radiotherapy and chemotherapy, the outcome of treatment in 17 children with nongerminomatous malignant germ cell tumor (NG-MGCT) was reviewed. The median follow-up period was 38 months after diagnosis, and the overall 3-year survival rate was 75%. Eleven patients who underwent craniospinal radiation (CSRT) did not receive chemotherapy. In 4 of them more than 90% of the tumor was removed, and they were free of disease at 16, 30, 93 and 111 months after surgery. Among the other 7, who did not undergo tumor resection (n=5) or had considerable residual tumor (n=2), 2 were disease-free at 73 and 88 months after diagnosis, and 5 died of recurrences. Of 6 patients who received cisplatin and etoposide chemotherapy in addition to CSRT, none showed intracranial recurrence, regardless of the extent of removal. The authors believe that multimodal treatment is the preferred choice and that chemotherapy plays an important role, especially when a significant amount of tumor remains after surgery. CSRT plays a major role at least in some patients. If chemotherapy is not feasible, radical removal plus CSRT seems to be an alternative. PMID- 10361970 TI - Endoscopy-assisted sural nerve harvest in infants. AB - A minimally invasive method of endoscopy-assisted sural nerve (SN) harvest in infants with obstetric brachial plexus lesions requiring nerve grafting procedures was applied to reduce the skin incision size and scarring at the donor site. Endoscopic visualization was achieved using a flexible and steerable Neuroview neuronavigational endoscope (Promedics, NL) 2.3 mm in diameter and 12 or 18 cm long in a peelaway sheath (700-9F) attached to a video camera. Through three 1.5-cm skin incisions the SN could be dissected free using a 2.5-mm diameter nerve stripper, pituitary curette or pituitary scissors under endoscopic vision from the opposite direction. To prevent any central nociceptive pain behavior the sural nerve was blocked by lidocaine, and sectioned first proximally in the popliteal fossa then distally at the lateral malleolus. PMID- 10361971 TI - Congenital glioblastoma diagnosed by fetal sonography. AB - Congenital brain tumors are very rare, and 2-9% of them are accounted for by glioblastomas. We encountered a case of congenital glioblastoma detected at the 39th week of gestation by fetal sonography, which revealed a large echogenic mass in the left temporo-parietal area of the fetal brain with significant midline shift and dilatation of the contralateral lateral ventricle. A detailed sonogram obtained 7 h later showed that the mass had increased in size, and this suggested an expanding hematoma. An emergency cesarean section was performed. Postnatal MRI demonstrated an enhancing mass with a large hematoma. Biopsy revealed a malignant brain tumor. Further management was refused and the boy died 6 days after birth. The postmortem pathological diagnosis was glioblastoma. When fetal sonography demonstrates an echogenic mass, a congenital brain tumor should be considered. The mode of delivery should be determined by the nature of the mass and the condition of the fetus. PMID- 10361972 TI - Dermal sinus and intramedullary spinal cord abscess. Report of two cases and review of the literature. AB - Intramedullary abscesses of the spinal cord are uncommon. Most of them occur in association with heart, pulmonary or urogenital infections. We report two cases of intramedullary spinal cord abscesses secondary to congenital dermal sinus. Only 14 cases of such an association have previously been reported. In our cases, dermal sinus was associated with an epidermoid tumour. The clinical presentation, pathogenesis, magnetic resonance imaging findings, surgical management and outcome are discussed. PMID- 10361973 TI - Anterior sacral meningocele completely occupied by an epidermoid tumor. AB - A 2-year-old girl presented with an anterior sacral meningocele completely occupied by an epidermoid tumor. Preoperative magnetic resonance imaging had shown the meningocele with contents of the same intensity as cerebrospinal fluid. Surgery via a posterior sacral approach disclosed the tumor beneath an unexpected membrane inside the meningocele. Additionally, the presence of pus inside epidermoid tumor suggested that possible episodes of asymptomatic meningitis or other infection might have occurred before treatment, these being the major complication in anterior sacral meningocele. Therefore, we recommend that surgical treatment should be performed at the earliest possible stage in childhood, once the diagnosis is established, and dural plasty carried out to prevent infectious complications. PMID- 10361974 TI - Clozapine: dopamine D1 receptor agonism in the prefrontal cortex as the code to decipher a Rosetta stone of antipsychotic drugs. AB - A large number of ligand binding studies have shown that clozapine has a number of receptor affinities, including those of the dopamine (DA) D1 and D2 receptor families. The study of intrinsic efficacy at these receptors is less straight forward. In the experiments summarised here, evidence is presented that clozapine behaves as an agonist at DA D1 receptors. Thus, the hypothermia produced by clozapine (2.5 mg kg(-1)) in the rat is fully antagonised by either of the selective DA D1 receptor antagonists SCH-23390 (0.1 mg kg(-1)) or NNC-687 (4 mg kg(-1)). These results provide an intriguing explanation for the clinical profile of clozapine as an atypical antipsychotic drug. Thus, there are supporting clinical and laboratory observations implicating DA D1 receptors in the prefrontal cortex in cognitive functions. Finally, clozapine displays features with regard to extrapyramidal motor mechanisms, and seizure thresholds, that could be explained by its properties as a DA D1 receptor agonist. PMID- 10361975 TI - The effect of in vitro exposure to white spirit on [Ca2+i] in synaptosomes from rats exposed prenatally to white spirit. AB - Female rats were exposed to white spirit (400 and 800 ppm for 6 hr/day) at day 7 20 during pregnancy. Thirty-five days after birth all female offspring were sacrificed, the brains removed, and the synaptosomal fractions prepared for in vitro studies. The cytosolic calcium concentration was measured using the FURA-2 technique. The results show that cytosolic calcium was increased in synaptosomes from rats exposed to white spirit prenatally compared to synaptosomes from unexposed rats. When synaptosomes were exposed to white spirit in vitro, the cytosolic calcium concentration changes were identical in all groups of rats. The membrane leakage measured as FURA-2 leakage from the synaptosomes identical in all three groups of animals. The results suggest that prenatal exposure to white spirit induces long-lasting and possibly irreversible changes in calcium homeostasis in the rat nervous system. PMID- 10361976 TI - Regulation of cytochrome P4501A metabolism by glutathione. AB - Gene expression of cytochrome P4501A (CYP1A) in the rainbow trout Oncorhynchus mykiss is dependent on aromatic hydrocarbon receptor signal transduction, and is markedly sensitive to tissue thiol status. Tissue glutathione (GSH) status was manipulated by exogenous GSH, L-buthionine-[S,R]-sulfoximine (BSO), lipoate or 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU). Tissue GSH contents were significantly elevated in GSH- and lipoate-supplemented trout. Hepatic, renal and plasma GSH levels were markedly arrested in BSO-treated trout. Oxidized glutathione (oxidized GSH) levels were significantly elevated in the BCNU supplemented group. Both BCNU treatment and BSO-induced GSH deficiency increased steady-state levels of hepatic CYPIA mRNA. Additional exposure to 0.1 mg/kg 3,3',4,4'-tetrachlorobiphenyl marginally suppressed the tetrachlorobiphenyl dependent CYP1A induction in BSO-treated livers compared with the respective thiol treatment groups. Tetrachlorobiphenyl exposures altered efficiencies of thiol treatments and increased oxidized GSH content in all but the BSO-treated groups. However, exposure to 5 mg/kg tetrachlorobiphenyl altered effects of thiol treatments on CYP1A mRNA to a small extent, but catalytic activity of CYP1A was many times suppressed in BSO-treated and lipoate-supplemented fish. These results suggest that thiol status interferes with CYPIA metabolism in a two-way mode of action and provide further evidence for a cross-talk between cytochrome P4501A and glutathione. PMID- 10361978 TI - In vitro cytotoxicity of the nitric oxide donor, S-nitroso-N-acetyl penicillamine, towards cells from human oral tissue. AB - The cytotoxicity of the nitric oxide donor, S-nitroso-N-acetyl-penicillamine (SNAP), towards cultured human cells from oral tissue was evaluated. The toxicity of SNAP to Smulow-Glickman gingival epithelial cells was correlated with the liberation of nitric oxide, as N-acetyl-D,L-penicillamine, the SNAP metabolites, N-acetyl-D,L-penicillamine disulfide and nitrite, and preincubated (denitrosylated) SNAP did not affect viability. Comparing equimolar concentrations of various nitric oxide donors, cytotoxicity appeared to be inversely related to the relative stability (i.e., half-life) of the test compound; the sequence of cytotoxicity for a 4 hr exposure was S nitrosoglutathione>>spermine NONOate> SNAP>DPTA NONOate>>DETA NONOate. Intracellular reduced glutathione (GSH) was lowered in S-G cells exposed to SNAP. Pretreatment of the cells with the GSH depleter, 1,3-bis-(chloroethyl)-1 nitrosourea (BCNU), enhanced the toxicity of SNAP Similar findings of enhanced sensitivity to SNAP were noted with gingival fibroblasts and periodontal ligament cells pretreated with BCNU. The toxicity of SNAP towards the gingival epithelial cells was decreased by cotreatment with the antioxidants, N-acetyl-L-cysteine, L ascorbic acid, and (+)-catechin. Cells exposed to SNAP exhibited nuclear aberrations, including multilobed nuclei and multinucleation. SNAP-induced cell death was apparently by apoptosis, as noted by fluorescence microscopy and DNA agarose gel electrophoresis. PMID- 10361977 TI - The effect of subcutaneous tetrathiomolybdate administration on copper and iron metabolism, including their regional redistribution in the brain, in the Long Evans Cinnamon rat, a bona fide animal model for Wilson's disease. AB - The present work was performed to examine the effect of tetrathiomolybdate on Cu and Fe metabolism, especially redistribution of Cu and Fe in the brains of Long Evans Cinnamon rats, with inherently abnormal Cu deposition in the liver. The drug was injected subcutaneously at 5 mg/kg of body weight twice a week for 65 days (total dose of 20 mg) into 40-day-old Long-Evans Cinnamon rats. In Long Evans Cinnamon rats treated with tetrathiomolybdate, the hepatic Cu concentration was 60 microg/g wet weight, compared to 170 microg/g in untreated rats. In seven brain regions (cerebellum, medulla oblongata, hypothalamus, striatum, midbrain, hippocampus and cortex) of the Long-Evans Cinnamon rats treated with tetrathiomolybdate. the Cu concentration (1.5 to 2.3 microg/g) was slightly lower (1.6 to 2.7 microg/g) than in untreated rats. A significant difference between the two groups was found only in the midbrain. Brain Fe concentrations in regions other than the striatum were not changed significantly by the tetrathiomolybdate injections. The hepatic Fe concentration was about 120 microg/g in Long-Evans Cinnamon rats without tetrathiomolybdate. Tetrathiomolybdate injection further increased the concentration to about 250 microg/g. Our results indicated that subcutaneous tetrathiomolybdate injection did not have an effect that stimulated redistribution of Cu and Fe in the seven brain regions examined, although hepatic Cu was markedly decreased and the removed Cu was deposited in kidneys, spleen and testes. The increased hepatic Fe level should be taken into account when considering side effects of the compound. PMID- 10361979 TI - The neuronal selective nitric oxide inhibitor AR-R 17477, blocks some effects of phencyclidine, while having no observable behavioural effects when given alone. AB - We have previously shown that the non-specific nitric oxide synthase inhibitor L NAME blocks the behavioural effects of phencyclidine, but not d-amphetamine. To characterise the specificity of these effects, we used the specific neuronal nitric oxide synthase inhibitor AR-R 17477 in two rat models of psychosis: the prepulse inhibition of the acoustic startle response and locomotor activity. In biochemical assays, AR-R 17477 was shown to be selective for the neuronal nitric oxide synthase isoform. Test drugs were given subcutaneously. AR-R 17477 (0.5, 1 and 5 mg/kg) antagonised phencyclidine-induced hyperlocomotion, while higher doses (10 and 20 mg/kg) were less efficaceous. AR-R 17477 (1 mg/kg) antagonised phencyclidine-induced deficit in prepulse inhibition of the acoustic startle response, while a higher dose (15 mg/kg) was less active. AR-R 17477 did not affect startle amplitude or prepulse inhibition of the acoustic startle response, did not affect locomotion and did not induce any changes in gross behaviour (sniffing, rearing, etc.) as determined in a subjective observation study. AR-R 17477 (1 mg/kg) did not alter the effect of d-amphetamine in prepulse inhibition of the acoustic startle response. Using radiotelemetry in rats, L-NAME (10 mg/kg subcutaneously) increased blood pressure and decreased heart rate while AR-R 17477 (10 mg/kg) did not have any significant effect on these parameters. The results show that a neuronal nitric oxide synthase inhibitor antagonises the effects of phencyclidine on prepulse inhibition of the acoustic startle response and locomotor activity, without exhibiting significant behavioural effects of its own and suggest that our earlier results with L-NAME depended upon an inhibition of neuronal nitric oxide synthase and not on an inhibition of endothelial nitric oxide synthase or inducible nitric oxide synthase. The observed effects are unlikely to be related to an effect on cardiovascular function. PMID- 10361980 TI - The relationship between activation of phosphoinositide-specific phospholipase C and growth stimulation by Ca2+-mobilizing hormones in hepatocytes. AB - Previous studies have shown that while vasopressin and angiotensin II are markedly more effective than norepinephrine and prostaglandin F2alpha in eliciting an acute elevation of inositol 1,4,5-trisphosphate (IP3), norepinephrine and prostaglandin F2alpha produce larger enhancement of DNA synthesis. This suggests that the initial activation of phosphoinositide-specific phospholipase C is not a common factor for the growth response to these agonists, but does not exclude a role of the integral of phospholipase C activity over a prolonged part of the prereplicative period, during which agonist-specific changes in responsiveness might occur. We show that vasopressin and angiotensin II also cause a prolonged elevation of cellular IP3 levels. which remain elevated for at least 60 min., while norepinephrine and prostaglandin F2alpha elevate IP3 levels slightly and transiently For vasopressin the dose-effect curves for IP3 accumulation and stimulation of DNA synthesis were closely parallel, while this was not the case for angiotensin II, norepinephrine, or prostaglandin F2alpha. After cultivation of the hepatocytes, hormone-stimulated IP3 accumulation rapidly declined, particularly in response to norepinephrine and prostaglandin F2alpha. When the IP3 response to norepinephrine and prostaglandin F2alpha was completely down-regulated, these agonists still enhanced the DNA synthesis. These results suggest that other mechanisms in addition to IP3 accumulation and Ca2+ release are likely to be involved in the growth stimulatory effects of the Ca2+ mobilizing agonists studied here, in particular for angiotensin II, norepinephrine, and prostaglandin F2alpha. PMID- 10361981 TI - Biology of presenilins as causative molecules for Alzheimer disease. AB - Many missense mutations in the presenilins are associated with autosomal dominant forms of familial Alzheimer disease (AD). Presenilin genes encode polytopic transmembrane proteins, which are processed by proteolytic cleavage and form high molecular-weight complexes under physiological conditions. The presenilins have been suggested to be functionally involved in developmental morphogenesis, apoptosis signal pathways, and processing of selected proteins including beta amyloid precursor protein. Although the underlying mechanism in which presenilin mutations lead to development of AD remains elusive, one consistent mutational effect is an overproduction of long-tailed amyloid beta-peptides. Furthermore, presenilins interact with beta-catenin to form presenilin complexes and presenilin mutations effect beta-catenin signalling pathways. PMID- 10361982 TI - Genetic landmarks through philately--a brief history of diabetes mellitus. PMID- 10361983 TI - The assent of a nation: genethics and Iceland. AB - The Icelandic parliament passed legislation authorizing the establishment of a national health sector database which will be sponsored financially by private enterprises through DeCode Genetics Inc. Health related data will be gathered from patients, without their informed consent, from all points of contact with Icelandic public and private health care providers. A centralized data curator will 'non-personalize' the identity of the subjects in a one-way coding system which the government and DeCode Genetics argue overrides the need for informed consent. This legislation is in conflict with the European Data Protection Act, which requires informed consent for the collection of personal data. The law raises many ethical questions regarding the central tenets of informed consent, the power of government, the rights of the human subject, and finally, the responsibility of the clinician balancing commitments of the patient and research. PMID- 10361984 TI - Autosomal dominant cataracts and Peters anomaly in a large Australian family. AB - Peters anomaly is a congenital corneal opacity with underlying defects in the posterior stroma, Descemets membrane and corneal endothelium. It is a disorder resulting from abnormal migration or function of neural crest cells and may include abnormalities of other anterior segment structures, such as the lens and iris. We report a family in which anterior segment abnormalities, including Peters anomaly and cataracts, were inherited in an autosomal dominant fashion. Although the PAX6 gene on chromosome 11 has been shown to be involved in some cases of anterior segment developmental defects, we found no evidence that the condition in this family is linked to the PAX6 gene. Identification of this gene will indicate another gene with major involvement in the development of the anterior segment of the eye. PMID- 10361985 TI - Linkage disequilibrium mapping of the Nova Scotia variant of Niemann-Pick disease. AB - Niemann-Pick type D (NPD) disease is a severe degenerative disorder of the nervous system characterized by the accumulation of tissue cholesterol and sphingomyelin. Because of a founder effect, it is unusually common in southwestern Nova Scotia, Canada. We have confirmed that almost all patients from 20 affected sibships descended on both sides from a small group of Acadians who settled in this region in about the year 1767. Previously using classic linkage analysis of this large kindred, we defined the critical gene region to a 13-cM chromosome segment between D18S869 and D18S66. Seven ESTs have been positioned within this interval. Carstea et al. (Niemann Pick C disease gene: homology to mediators of cholesterol homeostasis. Science 1997: 277: 232-235) recently demonstrated that one of these ESTs is the Niemann-Pick type C (NPCI) gene, the gene disrupted in most patients with NPC disease, and we have shown that a G3097- >T mutation in the NPC1 gene is also responsible for NPD. Here we report the development of five new polymorphic microsatellite markers and the testing for complete linkage disequilibrium in our single large NPD kindred that allowed us to reduce the NPD critical region to a 1-cM (1.3-1.6 Mb) interval between D18S1398 and D18S1108. In contrast, Carstea et al., using classic linkage analysis, required more than 18 unrelated NPC families to reduce the NPC1 critical region to a 5-cM interval. Our work supports the finding that NPD is an allelic variant of NPC1, and illustrates the power of large kindreds, which are common in Atlantic Canada and other relatively isolated areas, for gene mapping and identification. PMID- 10361986 TI - A proven case of materno-foetal transfusion determined by cytogenetic and DNA analysis. AB - We report a case of materno-foetal transfusion in a phenotypically normal male foetus after death in utero at the 35th week of gestation. We have used cytogenetic and polymerase chain reaction (PCR) microsatellite analysis to determine the presence of maternal cells in foetal blood collected by intracardiac puncture. In the intracardiac blood sample, maternal cells were estimated to comprise between 5 and 10% of nucleated foetal blood cells. When there is a suspicion of foetal genetic pathology, it is necessary to be aware that the foetal blood karyotype may be misrepresentative, as the analysed blood cells can indeed be of maternal origin. PMID- 10361987 TI - Procedure to protect confidentiality of familial data in community genetics and genomic research. AB - The collection of familial data is an essential step for community genetics programs or genetic research. Ethical issues concerning privacy and confidentiality present a major challenge in such programs. In order to keep familial data confidential, we have developed a family-based numerical coding procedure which allows the use of confidential data and the determination of familial relationships without risk of disclosure. This procedure is composed of two parts: the physical separation of identifying information and individual data; and the use of a code containing all the information required to build family trees. This procedure has been used in Eastern Quebec since 1995, mainly for screening, genetic counseling, research on familial dyslipidemias, public health intervention, and research projects on the genetics of complex traits, such as arterial hypertension and coronary artery disease. PMID- 10361988 TI - An unbalanced half-cryptic translocation involving the 6q subtelomeric region and 2p25.3 in a child with mental retardation: uses and limitations of fluorescence in situ hybridization. AB - We report on a 5-year-old boy with minor anomalies, growth retardation, and developmental delay carrying an extra chromatin material on the terminal band of the long arm of chromosome 6. To determine the origin of this extra material, whole chromosome fluorescence in situ hybridization (FISH) was used initially. Results showed fully painted 6qs, excluding the possibility of a derivative. However, maternal cytogenetic investigation suggested the presence of a possible half-cryptic balanced translocation that was further assessed using specific subtelomeric FISH probes of chromosome 6. Results showed that the 6q subtelomeric region was translocated on an A-group chromosome that was ultimately characterized, using FISH, as chromosome 2. This illustrates the use of specific subtelomeric regions and the limitations of whole chromosome FISH to identify the origin of a subtle chromosomal abnormality. PMID- 10361989 TI - Partial DiGeorge syndrome in two patients with a 10p rearrangement. AB - We describe 2 patients with a partial DiGeorge syndrome (facial dysmorphism, hypoparathyroidism, renal agenesis, mental retardation) and a rearrangement of chromosome 10p. The first patient carries a complex chromosomal rearrangement, with a reciprocal insertional translocation between the short arm of chromosome 10 and the long arm of chromosome 8, with karyotype 46, XY ins(8;10) (8pter 8q13::10p15-->10p14::8q24.1-->8qter) ins(10:8) (10pter--> 10p15::8q24.1- >8q13::10p14-->10qter). The karyotype of the second patient shows a terminal deletion of the short arm of chromosome 10. In both patients, the breakpoints on chromosome 10p reside outside the previously determined DiGeorge critical region II (DGCRII). This is in agreement with previous reports of patients with a terminal deletion of 10p with breakpoints distal to the DGCRII and renal malformations/hypoparathyroidism, and thus adds to evidence that these features may be caused by haploinsufficiency of one or more genes distal to the DGCRII. PMID- 10361990 TI - Clinical characteristics of Angelman syndrome patients with a non-IC-deleted imprinting mutation. PMID- 10361991 TI - Identification of a common N540K mutation in 8/18 Taiwanese hypochondroplasia patients: further evidence for genetic heterogeneity. PMID- 10361992 TI - Fok I polymorphism at the human vitamin D receptor gene locus in Europeans and Africans. PMID- 10361993 TI - Tetrasomy 21pter-q22.11: molecular, cytogenetic, and clinical findings. PMID- 10361994 TI - Spreading of metastases into cranial tumors: metastasis of a breast carcinoma to a pilocytic astrocytoma. AB - The case of a 29-year-old patient with metastasis of a breast carcinoma to a pilocytic astrocytoma is presented. Review of this exceptional occurrence is given. Discussing the possible causes of this coincidence we suggest that the phenomenon is probably an accidental event. PMID- 10361995 TI - Rapidly progressive multifocal leukoencephalopathy with substantial cell-mediated inflammatory response and with cognitive decline of non-Alzheimer type in a 75 year-old female patient. AB - Autopsy findings of rapidly progressive and widespread multifocal leukoencephalopathy (PML) in a 75-year-old woman with no known predisposing disease are demonstrated. Originally she was given a clinical working diagnosis of syndrome of progressive supranuclear palsy (PSP). The neuropathological investigation revealed widespread white and gray matter changes consistent with PML, and the JC virus was verified by EM, in situ hybridization and immunohistochemistry. In contrast to the few chronic inflammatory cells generally seen in PML in this case there was a substantial cell-mediated inflammatory response reflected in numerous T-helper and T-killer cells. The uncommon, widespread distribution of lesions and substantial cell-mediated response reported might indicate that the rearrangement of viral genome, previously suggested of importance for viral growth in the central nervous system (CNS), is also important for viral spread within the CNS, infectivity of glial cells and for the activation of cell-mediated immunity. PMID- 10361996 TI - Age-related adipose tissue mRNA expression of ADD1/SREBP1, PPARgamma, lipoprotein lipase, and GLUT4 glucose transporter in rhesus monkeys. AB - Aging has been shown to have an effect on the capacity to differentiate preadipocytes and on the expression of some genes expressed in adipose tissue. The mRNA levels of adipocyte differentiation-related genes were examined in rhesus monkeys (Macaca Mulatta) ranging in age from 7 to 30 years. The effect of aging on the expression of peroxisome proliferator activated receptor gamma (PPARgamma), adipocyte determination- and differentiation-dependent factor 1/sterol regulatory element binding protein 1 (ADD1/SREBP1), CCAAT/enhancer binding protein alpha (C/EBPalpha), lipoprotein lipase (LPL), GLUT4 glucose transporter, and adipsin were examined by slot blot analysis. Significant inverse correlations were observed between age and the mRNA levels of PPARgamma, ADD1/SREBP1, LPL, and GLUT4. The coordinate downregulation of these genes may be linked to the declining fat mass of senescent animals. There was no correlation between age and the mRNA levels of adipsin. The mRNA levels of these genes were not correlated to body weight orfasting plasma insulin. These findings indicate that aging may have an effect on the adipocyte differentiation program and this effect appears to be gene specific. PMID- 10361997 TI - Commentary: aging, differentiation-dependent gene expression, and fat cell function. PMID- 10361998 TI - The effects of nutritional manipulation and laboratory selection on lifespan in Drosophila melanogaster. AB - There are parallels between the effects of laboratory selection and nutritional manipulation on the expression of lifespan and other fitness-related characters in Drosophila melanogaster. However, little is known about the effects of laboratory selection and nutritional manipulation when applied simultaneously. Given that D. melanogaster is one of the major model organisms for testing theories of aging, simultaneous application of laboratory selection and nutritional manipulation is of considerable interest. To that end we developed six groups of five fold replicated populations selected for either early or late fertility. Each of these groups was maintained on either high- or low-nutrition diets. Comparisons among the groups showed that nutrition is neutral in selecting for lifespan. Moreover, the dietary-restriction response can be broken by simultaneous selection and nutritional manipulation. Finally, characters that respond in a parallel manner under selection or nutritional manipulation may not when the two are applied simultaneously. PMID- 10361999 TI - Effect of aging and dietary restriction on rat testicular germ cell apoptosis. AB - The purpose of this study was to determine whether apoptotic cell death of rat testicular germ cells varies with increased age or with dietary restriction. Slide-mounted cross-sections of formalin-fixed testis from 4-, 13-, and 23-month old ad-libitum-fed and 40% dietary-restricted male F344 rats were used in an in situ assay for apoptotic germ cells. Results show that in ad-libitum-fed rats, increased age caused a significant increase in the percentage of seminiferous tubules containing apoptotic cells, but a significant decrease in the number of apoptotic cells per apoptosis-positive tubule. In the 23-month-old rats, dietary restriction significantly increased both the percentage of apoptotic positive tubules and the number of apoptotic cells per apoptosis-positive tubule above ad libitum-fed values. Both ad-libitum and diet-restricted groups demonstrated a decrease in the percentage of apoptotic spermatogonia concomitant with a significant increase in the percentage of apoptotic primary spermatocytes with advanced age. The significant age-related increase in total numbers of apoptotic germ cells may reflect elimination of defective germ cells, which occurs with increased frequency in advanced age. Dietary restriction may induce enhanced screening against defective germ cells above that seen in the ad-libitum-fed rats. These results have potential implications in aging studies as well as research involving perturbation of normal germ cell homeostasis. PMID- 10362000 TI - Muscle quality and age: cross-sectional and longitudinal comparisons. AB - We addressed whether muscle quality (force per unit muscle mass) changes with age in cross-sectional and longitudinal analyses from three groups from the Baltimore Longitudinal Study of Aging: (1) Isometric arm strength studied cross-sectionally in 617 subjects with muscle mass estimated by cross-sectional area (CSA) from arm circumference and by 24-hour urinary creatinine excretion (CREAT); (2) longitudinal study for 10 to 25 years in 412 men using the same measures as the first group; and (3) isometric knee extensor strength studied cross-sectionally in 675 subjects; muscle mass estimated by CREAT, CSA from thigh circumference, and leg nonosseous fat free mass (FFM) from dual energy x-ray absorptiometry. Muscle quality declined in both arm and leg with age in cross-sectional analyses using CSA and FFM, but not CREAT. No age-associated arm muscle quality declines were observed longitudinally using CREAT or CSA. The relationship between muscle quality and age is dependent on how muscle mass is estimated and on whether subjects are studied cross-sectionally or longitudinally. In addition, CREAT may measure a muscle property not accounted for by CSA or FFM. PMID- 10362001 TI - Differential effects of aging on heart rate variability and blood pressure variability. AB - Previous studies investigating autonomic cardiovascular control in elderly persons usually included analysis of R-R interval but not of blood pressure variability. "Physiological" blood pressure rise during the aging process was not accounted for as a possible confounding factor. This study was designed to characterize the relationship between age and short-term heart rate (HR) and blood pressure (BP) variability, independently of the "physiological" rise in BP associated to aging. The study was carried out in 65 "normotensive" (BP< or =140/80 mm Hg) healthy subjects, ranging in age from 18 to 80 years. BP and HR were recorded at rest with a Finapres device. Low-frequency (LF = 0.066 to 0.129 Hz) and high-frequency (HF = respiratory peak +/-0.05 Hz) components of HR and BP variability were assessed using fast-Fourier spectral analysis. Transfer-function analysis between systolic BP and HR variability permitted the calculation of the gain of baroreflex sensitivity. Significant results of this study include a continuous and linear decline with age of normalized LF spectral power of HR in the standing position and of normalized HF spectral power of HR during paced breathing. No correlation was found between age and BP variability, except for LF diastolic BP spectral power in the standing position. The baroreflex gain was negatively correlated with age. The effect of aging on autonomic nervous system cardiac control is progressive and continuous throughout an 18-80 years age range. Although the aging process diminished HR variability and diastolic BP variability, it had no influence on systolic BP variability. PMID- 10362002 TI - Cardiovascular oscillations: from a physiological curiosity to a probe of aging and health? PMID- 10362003 TI - Aging alteration of cardiac vagosympathetic balance assessed through the tone entropy analysis. AB - BACKGROUND: Age-associated alterations of cardiac autonomic functions have been investigated intensively through heart rate variability analyses. However, changes with age in cardiac vagosympathetic balance remain to be elucidated. METHODS: We studied 142 male subjects (30-69 yr) at a health care center in Kyoto, Japan. Electrocardiographic data were collected from subjects in a recumbent position for 10 min in the morning. Analysis was done by classifying the subjects into four age groups. In a pharmacological experiment carried out in December 1996, tone was verified to reflect the cardiac vagosympathetic balance, and entropy the total autonomic neural efferent activity. We assessed the heart rate variability through the tone-entropy (T-E) analysis that was described previously. In essence, two indexes, tone and entropy, were defined on a distribution of successive variations of heart period. RESULTS: No significant differences were detected in clinical conditions among the four age groups. Tone increased and entropy decreased significantly with age. The aging process was expressed as a curvilinear path in T-E space. Compared to the pharmacological experiment, it was observed that aging degenerated the heart function from an ordinary to a denervated one. The same aging path was superimposed in the opposite direction on the heart recovery path after dynamic exercise in which cardiac vagosympathetic balance tended toward vagus division, corresponding with restoration of total autonomic activity. CONCLUSIONS: An age-related reduction in cardiac autonomic function was consistent with previous reports. The vagosympathetic balance was altered with this reduction: vagal predominance was impaired significantly in proportion to the withdrawal of total cardiac autonomic activity. PMID- 10362004 TI - Nursing home resident use of care directives. AB - BACKGROUND: The Patient Self-Determination Act of 1991 requires that nursing homes reimbursed by Medicare or Medicaid inform all residents upon admission of their rights to enact care directives in the event of terminal illness. This study investigated the relationship between care directive use and resident functional status. METHODS: We analyzed a version of the Minimum Data Set (MDS+) from a single state. We selected residents who were admitted to a nursing home in the first half of 1993 and followed them in the nursing home through the end of 1994. We created logistic models to examine independent correlates associated with having an advance directive or a do-not-resuscitate (DNR) order on admission. We then created similar logistic models to examine independent correlates associated with writing an advance directive or DNR order subsequent to admission. RESULTS: Of the 2,780 residents, 11% (292) had advance directives and 17% (466) had DNR orders upon admission. Of those without care directives upon admission, 6% (143) subsequently had an advance directive and 15% (339) subsequently had a DNR order. Cross-sectionally, older individuals and whites were more likely to have a care directive. Having poor cognitive and physical function was associated with having a DNR order upon admission. Longitudinally, longer stayers and whites were more likely to have an advance directive. Residents who lost physical function were more likely to have an advance directive and those who lost cognitive function were more likely to have a DNR order. CONCLUSIONS: Care directive use is influenced by a number of sociodemographic and functional characteristics. PMID- 10362005 TI - Reported and measured physical functioning in older inner-city diabetic African Americans. AB - BACKGROUND: The impact of diabetes on disability and physical functioning in older African Americans and potential causes of the excessive disability associated with diabetes in other studies have been inadequately investigated. METHODS: A population-based survey was performed comparing 116 self-reported diabetic inner-city African Americans aged 70 years and older to 522 nondiabetic persons from the same population. A subsample (n = 168) received a physical examination focused on body habitus, upper and lower body strength, balance, and timed physical performance tasks. Blood tests were obtained from 173 subjects. RESULTS: Diabetic individuals reported worse general health (p = .01), instrumental activities of daily living (p = .02), and modified versions of the Rosow-Breslau scale (p<.001) and the Stanford Health Assessment Questionnaire (p = .002). Diabetic persons also reported more falls (0.59 per person vs. 0.20, p = .019) and injurious falls (12% vs. 6%, p = .025). There were minimal differences in the strength, balance, and timed performance measures (analyzed separately by gender). In multivariable analyses, impairments in visual function and pain and light touch perception appeared to explain some of the association between diabetic status and poor general health, disability, and falls, with lesser contribution from the number of medical problems, number of medications, and glycemic control. CONCLUSIONS: Older inner-city diabetic blacks demonstrated worse general health, excess disability, and more falls compared to controls, although deficits in strength, balance, and timed performance could not be demonstrated. The cause of decreased functional status in diabetic elders deserves additional investigation, focusing especially on sensory function, glycemic control, and contribution from specific medical problems and medications. PMID- 10362006 TI - Sexual function in 1,202 aging males: differentiating aspects. AB - BACKGROUND: Late-life sexuality is an important quality-of-life issue that has been minimally explored. This survey seeks to extend our knowledge of the relationship of sexual attitudes and preferences to sexual functioning of a large group of older, community-dwelling men. METHODS: Older men aged 58-94 (N = 1,202) were surveyed with an anonymous self-administered questionnaire including 63 items regarding present and past, actual and desired sexual practices and attitudes. RESULTS: Although age correlated consistently with increased erectile dysfunction and decreased sexual activity, a substantial number of older men continued active sexual behaviors supported by positive attitudes toward sexual function. It was found that both health status and perceived partner's responsiveness are prominent moderators of the age effect. CONCLUSION: In the absence of social isolation and health issues, many older men show persistently active sexual lifestyles as evidenced in their interest and participation in sexual activities. These findings negate a portion of the starkly negative imagery of sexual expression in aging males. PMID- 10362007 TI - Exercise: effects on physical functional performance in independent older adults. AB - BACKGROUND: Age-related loss in physiologic capacities contributes to the decline in physical function in the elderly population. Despite the beneficial effects of exercise interventions on maximal physiologic capacity measures, the functional benefits have not been shown in independently living older adults. The objective of this study was to evaluate exercise in independent older adults for significant and meaningful improvements in physical function, not detected by commonly used measures of physical function. METHODS: In a randomized controlled study, 49 independently living men and women were assigned to either a nonexercise control group (Control; n = 26) or an exercise training group (Exercise; n = 23). Participants (age = 76+/-4) in good general health were recruited from retirement communities or apartments. The combined endurance and strength training was performed at 75% to 80% intensity; the groups met 3 times/week for 6 months of supervised sessions. Outcome measures included physical capacity, health status, and physical function using a newly developed performance test--the Continuous Scale-Physical Functional Performance test (CS PFP). RESULTS: Compared to the Control group, the Exercise group showed significant increases in maximal oxygen consumption (11%) and muscle strength (33%). No significant differences were found between groups for changes in the Sickness Impact Profile, SF-36 scales, or the 6-minute walk. However, the CS-PFP score improved significantly in the Exercise group (14%, effect size 0.80). CONCLUSIONS: Independent older adults gain meaningful functional benefits from several months of exercise training. The public health importance of physical activity may relate not just to its role in preventing decline, but also to its role in enhancing physical function. PMID- 10362008 TI - Older adults exhibit a reduced ability to fully activate their biceps brachii muscle. AB - BACKGROUND: Voluntary muscle strength declines significantly in older adults. One contributing factor to the strength loss is muscle atrophy developed in old age. Whether the ability to maximally activate the muscle decreases with age, however, is unknown. This study was intended to determine if the central nervous system command to maximally activate the biceps brachii muscle deteriorates with age. METHODS: Electrical stimulation pulses were applied to the skin overlying the biceps brachii muscle during maximal voluntary elbow-flexion contractions. The magnitude of force evoked on the maximal voluntary force was measured to determine the activation level (AL) of the muscle. RESULTS: The AL was 94% for the elderly group and 97% for the young group (100% AL indicates complete activation). The AL for both the elderly and young groups was significantly (p<.05) lower than 100%. The AL of the elderly group was significantly (p<.05) lower than that of the young group. CONCLUSIONS: The loss of voluntary strength in older adults is a mixed result of muscle atrophy and a reduced ability to fully activate muscle. PMID- 10362009 TI - The risk of hospitalization for acute myocardial infarction among older adults. AB - BACKGROUND: The purpose of this study was to prospectively examine the risk of hospitalization for acute myocardial infarction (AMI) in a large, nationally representative sample of very old men and women. METHODS: We utilized secondary analysis of the Longitudinal Study of Aging. Baseline (1984) in-person interview data were linked to Medicare hospitalization records for 1984-1991. Subjects were 6,071 noninstitutionalized adults 70 years old or older at baseline. Hospitalization for AMI was defined as having primary discharge diagnoses containing ICD9-CM 410 codes. Multivariable proportional hazards regression was used to evaluate the epidemiologic risks for all persons, and separately for women, men, self-respondents, those with no previous AMIs, and those with no history of coronary heart disease. RESULTS: Of the sample, 357 persons (5.9%; 172 women and 185 men) had at least one primary discharge diagnosis of AMI. Significant (p<.05) risk factors for being hospitalized with an AMI (adjusted hazards ratios in parentheses) from the pooled analysis were male gender (1.86), having no more than a grade school education (1.35), atherosclerosis (1.43), hypertension (1.29), coronary heart disease (1.63), angina (1.60), previous AMI (1.52), diabetes (1.89), and four or more lower body limitations (1.43). The gender-specific analyses, however, revealed that hypertension, angina, diabetes, and lower body limitations were risk factors only for women, and that having no more than a grade school education was a risk factor only for men. CONCLUSION: Men, especially those with low education, women with diabetes, angina, hypertension, or lower body limitations, and either men or women with previous AMIs, coronary heart disease, or atherosclerosis have elevated risks for AMI resulting in hospitalization, and they should be considered for evaluation and monitoring. Current protocols for therapeutic management should be adopted, and compliance should be encouraged. PMID- 10362010 TI - Preexisting medical conditions in adult day services: an examination of nonmetropolitan and metropolitan admissions. AB - BACKGROUND: It is not known what health conditions are being managed by day services staff because, to date, there is virtually no research on the types of preexisting medical conditions that clients bring to these community-based settings. Furthermore, it is not known whether or how nonmetropolitan clients differ from their metropolitan counterparts. METHODS: Census data for 1,448 individuals who were admitted to adult day services in Maryland during 1993 were examined. Variables were included for client characteristics, medical diagnoses at admission (based upon ICD-9-CM categories), and adult day center location (metropolitan vs. nonmetropolitan). RESULTS: Persons admitted to nonmetropolitan centers were more likely than those entering metropolitan centers to be diagnosed as having musculoskeletal, respiratory, cardiovascular, and endocrine conditions and as having a malignant neoplasm. Admissions to metropolitan centers were significantly more likely to be diagnosed with Alzheimer's disease. After using multiple logistic regression to control for a number of client characteristics, location of the facility remained significantly associated with all listed admission diagnoses except musculoskeletal conditions. CONCLUSION: Nonmetropolitan centers are caring for a distinctly different type of long-term care client than metropolitan centers, perhaps because few other long-term care options are available to families in sparsely populated settings. The differences in the medical conditions of their clients should affect most aspects of the day services program, including budgets for appropriate professional staff, staff qualifications, resources for client/family education and counseling, and expected outcomes. PMID- 10362011 TI - Hospitalization and Alzheimer's disease: results from a community-based study. AB - BACKGROUND: Prior studies offer conflicting findings on whether Alzheimer's disease (AD) is associated with an increased risk of hospitalization. METHODS: We investigated AD and hospitalization in the Washington Heights-Inwood Columbia Aging Project (WHICAP), a community-based study of 2,334 elders in New York City. In 1996, an electronic medical records system was established that allows an e mail alert to be sent to the research team whenever WHICAP subjects are admitted to Columbia-Presbyterian Medical Center (CPMC), the site of hospital care for the majority of subjects. RESULTS: Of the WHICAP cohort, 13.1% was admitted to CPMC in 21 months of follow-up; 17.5% of AD patients and 11.9% of unaffected subjects were admitted (p<.01). Multivariate logistic regression models showed that more advanced AD (Clinical Dementia Rating scale 3+) was a significant risk factor for hospitalization independently of age, gender, education, comorbid medical conditions, and death in the follow-up period (OR 2.3; 95% CI: 1.1, 4.6); subjects with mild or moderate AD did not show a significantly elevated risk. The prevalence of psychiatric symptoms did not differ between AD subjects who were hospitalized in the reporting period and AD subjects who were not hospitalized. Infectious disease was a more common discharge diagnosis for subjects with AD (p<.05). CONCLUSIONS: In this community-based cohort, subjects with severe AD were more likely to be hospitalized than unaffected subjects. The increased use of hospital care by these AD patients appears to be specific to AD but is not a result of psychiatric morbidity or end-of-life care. Rather, a greater risk of medical complications that require hospital care, especially infections, appears to be characteristic of severe AD. PMID- 10362012 TI - Myostatin, a transforming growth factor-beta superfamily member, is expressed in heart muscle and is upregulated in cardiomyocytes after infarct. AB - Myostatin is a secreted growth and differentiating factor (GDF-8) that belongs to the transforming growth factor-beta (TGF-beta) superfamily. Targeted disruption of the myostatin gene in mice and a mutation in the third exon of the myostatin gene in double-muscled Belgian Blue cattle breed result in skeletal muscle hyperplasia. Hence, myostatin has been shown to be involved in the regulation of skeletal muscle mass in both mice and cattle. Previous published reports utilizing Northern hybridization had shown that myostatin expression was seen exclusively in skeletal muscle. A significantly lower level of myostatin mRNA was also reported in adipose tissue. Using a sensitive reverse transcription polymerase chain reaction (RT-PCR) technique and Western blotting with anti myostatin antibodies, we show that myostatin mRNA and protein are not restricted to skeletal muscle. We also show that myostatin expression is detected in the muscle of both fetal and adult hearts. Sequence analysis reveals that the Belgian Blue heart myostatin cDNA sequence contains an 11 nucleotide deletion in the third exon that causes a frameshift that eliminates virtually all of the mature, active region of the protein. Anti-myostatin immunostaining on heart sections also demonstrates that myostatin protein is localized in Purkinje fibers and cardiomyocytes in heart tissue. Furthermore, following myocardial infarction, myostatin expression is upregulated in the cardiomyocytes surrounding the infarct area. Given that myostatin is expressed in fetal and adult hearts and that myostatin expression is upregulated in cardiomyocytes after the infarction, myostatin could play an important role in cardiac development and physiology. PMID- 10362013 TI - Role of telomerase in cellular proliferation and cancer. AB - Telomerase is a cellular reverse transcriptase that helps to provide genomic stability in highly proliferative normal, immortal, and tumor cells by maintaining the integrity of the chromosome ends, the telomeres. The activity of telomerase is associated with the majority of malignant human cancers. Telomerase or another mechanism for telomere maintenance is required for continuous tumor cell proliferation. Telomerase-positive cells that exit the cell cycle via quiescence downregulate telomerase through a transcriptional repression pathway. In the case of cell cycle exit via terminal differentiation, proteolysis of telomerase may also be involved. In response to mitogenic or growth factor signaling, telomerase-competent quiescent cells reenter the cell cycle and express telomerase activity independent of DNA synthesis. Under normal growth conditions, inhibition of telomerase activity in tumor-derived cells results in continued cell division coupled with telomere shortening, eventually followed by cellular senescence or death. Thus, repression of telomerase activity may be a novel adjuvant therapy for the treatment of human cancer and detection of telomerase activity may be important for cancer diagnostics. PMID- 10362014 TI - Microvillar Ca++ signaling: a new view of an old problem. AB - Proceeding from the recent finding that the main components of the Ca++ signal pathway are located in small membrane protrusions on the surface of differentiated cells, called microvilli, a novel concept of cellular Ca++ signaling was developed. The main features of this concept can be summarized as follows: Microvilli are formed on the cell surface of differentiating or resting cells from exocytic membrane domains, growing out from the cell surface by elongation of an internal bundle of actin filaments. The microvillar tip membranes contain all functional important proteins synthesized such as ion channels and transporters for energy-providing substrates and structural components, which are, in rapidly growing undifferentiated cells, distributed over the whole cell surface by lateral diffusion. The microvillar shaft structure, a bundle of actin filaments, forms a dense cytoskeletal matrix tightly covered by the microvillar lipid membrane and represents an effective diffusion barrier separating the microvillar tip compartment (entrance compartment) from the cytoplasm. This diffusion barrier prevents the passage of low molecular components such as Ca++ glucose and other relevant substrates from the entrance compartment into the cytoplasm. The effectiveness of the actin-based diffusion barrier is modulated by various signal pathways and effectors, most importantly, by the actin-depolymerizing/reorganizing activity of the phospholipase C (PLC) coupled Ca++ signaling. Moreover, the microvillar bundle of actin filaments plays a dual role in Ca++ signaling. It combines the function of a diffusion barrier, preventing Ca++ influx into the resting cell, with that of a high-affinity, ATP dependent, and IP3-sensitive Ca++ store. Activation of Ca++ signaling via PLC coupled receptors simultaneously empties Ca++ stores and activates the influx of external Ca++. The presented concept of Ca++ signaling is compatible with all established data on Ca++ signaling. Properties of Ca++ signaling, that could not be reconciled with the basic principles of the current hypothesis, are intrinsic properties of the new concept. Quantal Ca++ release, Ca(++)-induced Ca++ release (CICR), the coupling phenomen between the filling state of the Ca++ store and the activity of the Ca++ influx pathway, as well as the various yet unexplained complex kinetics of Ca++ uptake and release can be explained on a common mechanistic basis. PMID- 10362015 TI - Bone morphogenetic protein-2 acts synergistically with transforming growth factor beta3 during endothelial-mesenchymal transformation in the developing chick heart. AB - In the early embryonic heart, endothelial cells in atrioventricular (AV) and outflow tract (OT) regions are transformed into the invasive mesenchymal cells that form endocardial cushion tissue (endothelial-mesenchymal transformation). It has been reported that bone morphogenetic proteins (BMPs) are transcribed in the AV and OT regions of the embryonic mouse heart. We previously reported that transforming growth factor beta 3 (TGFbeta3) triggers the initial phenotypic changes seen in endothelial-mesenchymal transformation. We cloned BMP2 from embryonic chick hearts and examined its functional role during endocardial cushion tissue formation. In situ hybridization showed BMP2 transcripts in the myocardium of the AV and OT regions, but not in endothelial/mesenchymal cells. Antisense oligodeoxynucleotides to BMP2 inhibited mesenchyme formation in AV endocardium cocultured with associated myocardium. This inhibitory effect was reversed by the addition of recombinant BMP2. In cultured AV endothelial monolayers, recombinant BMP2 did not induce any cellular phenotypic changes characteristic of endothelial-mesenchymal transformation. However, BMP2 enhanced the TGFbeta-induced initial phenotypic changes associated with endothelial mesenchymal transformation. These results suggest that BMP2 1) plays an important role in the formation of endocardial cushion tissue and 2) acts synergistically with TGFbeta3 in the regulation of this developmental event. PMID- 10362016 TI - Telomerase activation in human fibroblasts during escape from crisis. AB - The immortalization of human diploid fibroblasts requires the circumvention of both the senescence (M1) and crisis (M2) mechanisms of growth control. Cells expressing the SV40 T antigen virtually always bypass senescence, but only rarely escape crisis. The low frequency of this latter event indicates that cellular mutations are necessary to escape crisis. Thirteen subpopulations of T antigen expressing human fibroblasts were cultured into crisis. Colonies that appeared to resume growth were assayed for telomerase activity, telomere maintenance, and the immortal phenotype. Our results show that 33 of 35 colonies were telomerase negative and were not immortal. Two colonies were telomerase positive when assayed in the first approximately 15 population doublings after crisis. The first was strongly positive, maintained telomeres at a stable short length, and was later determined to be immortal. The second initially had a weak telomerase signal, grew extremely slowly, and when examined had greatly elongated telomeres consistent with the ALT (alternative lengthening of telomeres) mechanism of telomere maintenance. These cells eventually grew faster and were later determined to be immortal. Additionally, two subpopulations had initially weak and later strong telomerase activity and the cells never entered a defined crisis period. We observed a perfect correlation between telomere maintenance and escape from crisis, supporting the hypothesis that the lack of stable telomeres causes crisis and that the ability to maintain telomeres abrogates crisis. PMID- 10362017 TI - FGF9 is an autocrine and paracrine prostatic growth factor expressed by prostatic stromal cells. AB - Polypeptide growth factors, including members of the fibroblast growth factor (FGF) family, play an important role in the growth and maintenance of the normal prostate. We have found that FGF9 is expressed at high levels in the normal peripheral and transition zone of the human prostate. Analysis of FGF9 production by primary cultures of prostatic epithelial and stromal cells has shown that FGF9 is produced and secreted by the prostatic stromal cells. Neither of these processes appears to be modulated by androgens. Production of FGF9 by stromal cells in vivo was confirmed by immunohistochemistry. FGF9 is a potent mitogen for both prostatic epithelial and stromal cells in culture and is a more potent mitogen for these cells than either FGF2 or FGF7, two other FGFs expressed in the human prostate. FGF9 is an abundant secreted growth factor that can act as both a paracrine mitogen for epithelial cells and an autocrine mitogen for stromal cells. Western blot analysis of tissue extracts from the normal and hyperplastic transition zone shows that FGF9 is present at two to threefold higher levels in the hyperplastic transition zone. Overexpression of this paracrine and autocrine growth factor may play an important role in the epithelial and stromal proliferation in benign prostatic hyperplasia. PMID- 10362018 TI - Insulin-like growth factor-II/mannose 6 phosphate receptors facilitate the matrix effects of latent transforming growth factor-beta1 released from genetically modified keratinocytes in a fibroblast/keratinocyte co-culture system. AB - This study was conducted to explore the mechanism of activation of transforming growth factor-beta1 (TGF-beta1) which is critical to its role in many physiological and pathological conditions. To date, almost all reports concerning TGF-beta1 activation delineated that release of mature TGF-beta1 from latency associated protein (LAP) is required for its activation. We report that latent TGF-beta1 (LTGF-beta1) released from TGF-beta1 genetically modified keratinocytes grown in the top chamber of a co-culture system functions as a fibrogenic factor through interaction with insulin-like growth factor-II/mannose-6-phosphate (IGF II/M6P) receptors of human dermal fibroblasts grown in the lower chamber of this system. Following successful transduction, the pLin-LTGF-beta1 vector was amplified in PA31 7 packaging cells which possess viral structural proteins for vector in the presence of neomycin. Conditioned medium derived from packaging cells containing competent viral particles was then used to transduce either keratinocytes or fibroblasts grown in the upper chamber of a co-culture system, in which a 0.4 microm porous membrane separates the two chambers. In this way, LTGF-beta1 produced by transduced cells in the upper chamber is released and diffuses into the lower chambers where dermal fibroblasts are grown. Conditioned medium from the lower chamber was removed 3 days later and used to evaluate the latency and bioactivity of TGF-beta1 using enzyme-linked immunosorbent assay (ELISA) and mink lung (Mv1 Lu) epithelial growth inhibition assay. Cells were also harvested and used for RNA extraction. The results of these experiments showed that 1) the TGF-beta1-LAP complex, which was latent in traditionally used mink lung growth inhibition assay, directly modulated the expression of collagenase, type I, and type III collagen mRNA by dermal fibroblasts; 2) this stimulation was inhibited by M6P in a dose-dependent manner; 3) the TGF-beta1-LAP inhibits Mv1Lu epithelial cells only when this complex was incubated with cell membranes isolated from dermal fibroblasts; and 4) LTGF-beta1 activation seems to occur through a conformational alteration rather than by release of the mature TGF-beta1 from LAP in our co-cultured system. This conformational alteration seems to occur through the interaction of the TGF-beta1-LAP complex with the IGF II/M6P receptors. Thus, the quantity of IGF-II/M6P receptors is important in cellular response to LTGF-beta1 in any physiological and pathological conditions. PMID- 10362019 TI - Activation of the cAMP signaling pathway increases apoptosis in human B-precursor cells and is associated with downregulation of Mcl-1 expression. AB - During B- and T-cell ontogeny, extensive apoptosis occurs at distinct stages of development. Agents that increase intracellular levels of cAMP induce apoptosis in thymocytes and mature B cells, prompting us to investigate the role of cAMP signaling in human CD10+ B-precursor cells. We show for the first time that forskolin (which increases intracellular levels of cAMP) increases apoptosis in the CD10- cells in a dose-dependent manner (19%-94% with 0-1,000 microM forskolin after 48 hours incubation, IC50 = 150 microM). High levels of apoptosis were also obtained by exposing the cells to the cAMP analogue 8-chlorophenylthio-cAMP (8 CPT-cAMP). Specific involvement of cAMP-dependent protein kinase (PKA) was demonstrated by the ability of a cAMP antagonist, Rp-isomer of 8-bromo-adenosine- 3', 5'- monophosphorothioate (Rp-8-Br-cAMPS), to reverse the apoptosis increasing effect of the complementary cAMP agonist, Sp-8-Br-cAMPS. Furthermore, we investigated the expression of Bcl-2 family proteins. We found that treatment of the cells with forskolin or 8-CPT-cAMP for 48 hours resulted in a fourfold decline in the expression of Mcl-1 (n = 6, P = 0.002) compared to control cells. The expression of Bcl-2, Bcl-xL, or Bax was largely unaffected. Mature peripheral blood B cells showed a smaller increase in the percentage of apoptotic cells in response to 8-CPT-cAMP (1.3-fold, n = 6, P = 0.045) compared to B-precursor cells, and a smaller decrease in Mcl-1 levels (1.5-fold, n = 4, P = 0.014). Taken together, these findings show that cAMP is important in the regulation of apoptosis in B-progenitor and mature B cells and suggest that cAMP-increased apoptosis could be mediated, at least in part, by a decrease in Mcl-1 levels. PMID- 10362020 TI - Hepatocyte growth factor induces tubulogenesis of primary renal proximal tubular epithelial cells. AB - Hepatocyte growth factor (HGF)-induced tubulogenesis has been demonstrated with renal epithelial cell lines grown in collagen gels but not with primary cultured renal proximal tubular epithelial cells (RPTEs). We show that HGF selectively induces proliferation and branching morphogenesis of primary cultured rat RPTEs. Additional growth factors including fibroblast growth factor (FGF)-1, epidermal growth factor (EGF), FGF-7, or insulin-like growth factor-1 (IGF-1) did not selectively induce tubulogenesis. However, when administered in combination, these factors initiated branching morphogenesis comparable to HGF alone and greatly augmented HGF-induced proliferation and branching. Microscopic analysis revealed that branching RPTEs were undergoing tubulogenesis and formed a polarized epithelium. TGF-beta1 blocked HGF- or growth factor cocktail (GFC; HGF, FGF-1, EGF, IGF-1)-induced proliferation and branching morphogenesis. Adding TGF beta1 after GFC-induced tubulogenesis had occurred caused a progressive regression of the tubular structures, a response associated with an increase in apoptosis of the RPTEs. Primary cultured RPTEs are capable of undergoing HGF induced tubulogenesis. Unlike cell lines, combinations of growth factors differentially augment the response. PMID- 10362021 TI - Changes in mitochondrial mass, membrane potential, and cellular adenosine triphosphate content during the cell cycle of human leukemic (HL-60) cells. AB - Oxidative phosphorylation within the inner mitochondrial membrane generates the majority of cellular adenosine triphosphate (ATP) required for normal physiological functions (including regulation of cell volume and solute concentration, maintenance of cellular architecture, and synthesis of essential macromolecules). Its efficient functioning depends on the maintenance of an electrochemical gradient and is tightly coupled to the energetic demands of the cell and/or tissue. Commitment to and completion of the cell division cycle are sensitive to changes in the availability of mitochondrially derived ATP, although the relationship between cell cycle and mitochondrial physiology is poorly understood. Using vital, mitochondrial-specific fluorochromes to differentiate between mitochondrial mass (10-N-nonyl acridine orange) and mitochondrial membrane potential (Rhodamine 123), together with a quantification of total cellular ATP levels, it was possible to generate profiles of these mitochondrial characteristics in HL-60 cells at different stages of their cell cycle. The data suggest that the availability of ATP changes in a cell cycle-specific manner and cannot be predicted by changes in mitochondrial mass or membrane potential. Furthermore, transition points in the cell cycle where ATP availability is low with respect to the amount of functional inner mitochondrial membrane have been observed. We suggest that these cell cycle phase transitions are sensitive to inhibition of mitochondrial activity because the basal levels of available ATP at these points are nearer to a theoretical "minimal threshold" below which cell cycle progression is inhibited. PMID- 10362022 TI - Glucose-regulated stresses cause degradation of DNA topoisomerase IIalpha by inducing nuclear proteasome during G1 cell cycle arrest in cancer cells. AB - The glucose-regulated stress response of cancer cells leads to a decreased expression of DNA topoisomerase IIalpha (topo IIalpha) and a cell cycle arrest at the G1 phase. In this study, we found that the topo IIalpha decrease occurred specifically during the G1 arrest in human colon adenocarcinoma HT-29 cells. The intracelluar level of topo IIalpha in HT-29 cells was relatively constant regardless of cell cycle position in the exponentially growing state, determined using a centrifugal elutriation technique and synchronizing the cells with a mitotic inhibitor nocodazole. Interestingly, when the cell cycle was arrested in the M phase by nocodazole, the topo IIalpha level remained high even in stressed cells. After the stressed cells were released from the M phase, topo IIalpha steeply decreased along with cell cycle progression followed by the next G1 arrest. This decrease in nuclear topo IIalpha protein was completely inhibited by selective inhibitors for proteasome. Furthermore, we found that proteasome activity was elevated three to fourfold in the nuclear extract of stressed cells over unstressed cells. Accordingly, there were increased amounts of nuclear proteasome subunits, although total intracellular content of the subunits did not change in stressed cells. These findings indicate that the expression of topo IIalpha in stressed cells is downregulated at the G1 phase by proteasome-mediated degradation and that the proteolysis of topo IIalpha can be facilitated by the nuclear accumulation of proteasome. PMID- 10362023 TI - Differential metal response and regulation of human heavy metal-inducible genes. AB - A number of heavy metal-inducible genes have been reported, but their ranges of response to various metal species are not well known. It is also unclear if these genes are regulated through common mechanisms. To answer these questions, we compared induction kinetics of human metal-inducible genes including the MT-IIA (coding for a metallothionein isoform), hsp70 (coding for the 70-kDa heat-shock protein), and c-fos genes in HeLa cells exposed to Zn, Cd, Ag, Hg, Cu(II), Co, or Ni ions. Transcripts from these three genes were increased after exposure to wide ranges of metals, but each gene was unique in its induction kinetics. Generally, induction was observed at lower metal concentrations in the order of MT-IIA, hsp70, and c-fos. These genes also showed differential responses in time course: more rapid induction was observed in the order of c-fos, hsp70, and MT-IIA after exposure to Zn or Cd. Since the metal-responsive element (MRE) and heat shock element (HSE) of the MT-IIA and hsp70 genes, respectively, are thought to be the cis-acting DNA elements that mediate metal response, we compared the properties of proteins that specifically bind to these elements. MRE-binding activity was detected only in the extract from cells exposed to Zn. By contrast, HSE-binding activity was detected in extracts from cells treated with Zn, Cd, Ag, and Cu. The former was also activated by Zn in vitro, while the latter was not. Each of these DNA-binding activities showed no affinity to the recognition sequence of the other. These results demonstrate that the human metal-inducible genes have broad ranges of response to a variety of heavy metals, but suggest that they are probably regulated through independent mechanisms. PMID- 10362024 TI - Differences in electron transport potential, antioxidant defenses, and oxidant generation in young and senescent fetal lung fibroblasts (WI-38). AB - The activities and mRNA abundances of enzymes that regulate the rate of electron flow through the electron transport chain (ETC), including NADH dehydrogenase, succinate dehydrogenase, and cytochrome c oxidase, were examined in young and senescent fetal lung fibroblasts (WI-38). We also determined the activities and mRNA abundances of antioxidant defenses including superoxide dismutase, catalase, and glutathione peroxidase. We confirmed our previous report of a senescence related increase in the abundance of ND4, a mitochondrially encoded subunit of NADH dehydrogenase. The activities of cytochrome c oxidase and NADH dehydrogenase were also elevated in senescent cultures. No differences were observed in the mRNA abundances of COX-1, a mitochondrially encoded subunit of cytochrome c oxidase or of nuclearly encoded subunits of various electron transport components (SD, COX-4, and ND 51). Lucigenin-detected chemiluminescence and H2O2 generation were both elevated in senescent cells. Catalase activity was also elevated in senescent fibroblasts. However, no differences in catalase mRNA abundance were observed. A small decrease in GSH peroxidase (GPx) mRNA abundance was observed in senescent cells. No other changes in the activities or mRNA abundances of any of the antioxidant defenses were observed in early and late passage cultures. The relationships between oxidant generation, mitochondrial enzyme activities, and antioxidant defense observed during proliferative senescence are dissimilar to those detected between fetal and postnatal fibroblasts as well as those found between fibroblast lines obtained from young and old individuals. The relevance of the differences between these models is discussed. PMID- 10362025 TI - Cellular proliferation potential during aging and caloric restriction in rhesus monkeys (Macaca mulatta). AB - Caloric restriction (CR) is the most successful method of extending both median and maximal lifespans in rodents and other short-lived species. It is not yet clear whether this method of life extension will be successful in longer-lived species, possibly including humans; however, trials in rhesus monkeys are underway. We have examined the cellular proliferative potential of cells from CR and AL (ad libitum fed) monkey skin cells using two different bioassays: colony size analysis (CSA) of dermal fibroblasts isolated and cloned directly from the skin and beta-galactosidase staining at pH 6.0 (BG-6.0) of epidermal cells in frozen sections of skin. Decreases in both proliferative markers occurred with age, but no differences were observed between CR and AL animals. Skin biopsies were obtained from AL and CR rhesus monkeys from two different aging colonies, one at the National Institute on Aging (NIA) and one at the University of Maryland-Baltimore (UMB). These biopsies were used as a source of tissue sections and cells for two biomarkers of aging assays. The CR monkeys had been maintained for 9-12 years on approximately 70% of the caloric intake of control AL animals. In the CSA studies, the fraction of small clones increased significantly and the fraction of large clones decreased significantly with increasing age in AL monkeys. The frequency of epidermal BG-6.0 staining cells increased with age in older (>22 years) AL monkeys, but most predominately in those of the UMB colony, which were somewhat heavier than the NIH AL controls. Old monkeys on CR tended to have fewer BG-6.0-positive cells relative to old AL-derived epidermis, but this effect was not significant. These results indicate that cellular proliferative potential declined with age in Macaca mulatta, but was not significantly altered by CR under these conditions. Although these experiments are consistent with an absence of effect of CR on monkey skin cell proliferative potential, we have found in previous experiments with mice that a longer duration of CR (as a fraction of total lifespan) was needed to demonstrate CR-related improvement in clone size in mice. Further studies on the now mid-aged monkeys will be needed as their age exceeds 20 years to conclusively rule out an effect of CR on proliferative potential of skin cells from these primates. PMID- 10362027 TI - Fatigue and sarcoidosis. PMID- 10362026 TI - Biosynthesis of alpha2(IV) and alpha1(IV) chains of collagen IV and interactions with matrix metalloproteinase-9. AB - In vitro binding studies with latent matrix metalloproteinase-9 (pro-MMP-9) have revealed the existence of nondisulfide-bonded alpha2(IV) chains on the cell surface capable of forming a high-affinity complex with the enzyme. Here we investigated the biosynthesis and cellular distribution of alpha2(IV) and alpha1(IV) chains in breast epithelial (MCF10A and MDA-MB-231) and fibrosarcoma (HT1080) cells by pulse-chase analysis followed by immunoprecipitation with chain specific monoclonal antibodies (mAb). These studies showed that whereas the alpha1(IV) chain remained in the intracellular compartment, nondisulfide-bonded alpha2(IV) chains were secreted into the media in a stable form. Consistently, only alpha2(IV) was detected on the cell surface by surface biotinylation or indirect immunofluorescence. In agreement with the pulse-chase analysis, media subjected to co-precipitation experiments with pro-MMP-9 or pro-MMP-9-affinity chromatography followed by immunoblotting with chain-specific mAbs resulted in the detection of alpha2(IV). A preferential secretion of nondisulfide-bonded alpha2(IV) chains was also observed in CHO-K1 cells transiently transfected with full-length mouse alpha2(IV) or alpha (IV) cDNAs. However, a complex of mouse alpha1(IV) with pro-MMP-9 was coprecipitated with exogenous enzyme from lysates of CHO-K1 cells transfected with mouse alpha1(IV), suggesting that under overexpression conditions the enzyme can also interact with the alpha1 (IV) chain. Collectively, these studies further demonstrate the interactions of pro MMP-9 with collagen IV chains and a unique processing and targeting of nondisulfide-bonded alpha2(IV) chains that may play a role in the surface/matrix association of pro-MMP-9. PMID- 10362028 TI - Radiation-induced injury in the "nonirradiated" lung. PMID- 10362029 TI - Association of fatigue with an acute phase response in sarcoidosis. AB - The pathophysiological explanation for fatigue, one of the most common symptoms in sarcoidosis, still has to be elucidated. It was hypothesized that the presence of fatigue is associated with an acute phase response in sarcoidosis. A cross sectional study was performed in 38 sarcoidosis patients. Resting energy expenditure (REE) was measured in the fasting state by indirect calorimetry using a ventilated hood and adjusted for fat-free mass (FFM). Patients with fatigue (n=25) also suffered more frequently from other symptoms, such as exercise intolerance (p=0.01), the need for sleep (p=0.02) and weight loss (p=0.01), compared to those without fatigue (n=13). However, no relationship was found between fatigue and serum angiotensin-converting enzyme (sACE) or lung function impairment. Patients with fatigue had higher levels of C-reactive protein (CRP) (11.4+/-6.8 microg x mL(-1), p<0.0001) and REE adjusted for FFM (33.0+/-3.7 kcal x kg FFM(-1), p<0.003) compared to those without fatigue (3.2+/-2.2 mg x mL(-1); 29.2+/-2.8 kcal x kg FF(-1)). Furthermore, REE/FFM was significantly related to CRP (r=0.54, p=0.001). This study confirms the presence of an acute phase response as indicated by metabolic derangements and a moderate increase in C reactive protein levels in sarcoidosis, particularly in those patients with constitutional symptoms. Future studies should focus on the clinical relevance and therapeutic implications of these findings. PMID- 10362030 TI - The angiotensin-converting enzyme DD gene is associated with poor prognosis in Finnish sarcoidosis patients. AB - Angiotensin-converting enzyme (ACE) genotypes may reflect prognosis in sarcoidosis. They were determined in 59 Finnish sarcoidosis patients and 70 healthy control subjects. The prognosis of the sarcoidosis patients was determined after follow-up for 1, 2, 3, 5 and >5 yrs and classified as good (normal chest radiograph and lung function, no signs of extrapulmonary disease activity within 2 yrs from diagnosis), intermediate (neither good nor poor) or poor (persisting unstable pulmonary infiltrates, vital capacity and diffusing capacity of the lung for carbon monoxide <50% predicted and/or extrapulmonary disease activity after >5 yrs follow-up). The DD, ID and II genotypes were found in 31 and 27%, in 54 and 49%, and in 15 and 24% of patients and control subjects respectively. The odds ratio (DD+ID to II) was 1.45 (95% confidence interval 0.60 3.49). The D alelle was found more often in patients (58%) and in control subjects (51%) than the I allele but the difference was not statistically significant. Statistically significantly more patients with the DD genotype had a poor prognosis compared with patients with II homozygotes and ID heterozygotes. Among 11 patients with Lofgren's syndrome (bilateral hilar lymphadenopathy and erythema nodosum), four had the DD genotype. Three of these patients had a prognosis despite presenting a clinical picture usually associated with a good prognosis. The angiotensin-converting enzyme genotype may be a prognostic marker in sarcoidosis and larger studies are warranted to define its clinical utility. PMID- 10362031 TI - Bilateral lymphocytic alveolitis: a common reaction after unilateral thoracic irradiation. AB - The main aim of the present study was to assess the early diagnostic value of bronchoalveolar lavage (BAL) in radiation-induced lung injury in patients with breast carcinoma. Twenty-six females receiving postoperative radiotherapy for breast cancer were evaluated before and 0, 15, 30, 60, and 180 days after radiotherapy. History, physical examination, chest radiographs, and pulmonary function tests were obtained. BAL, including lymphocyte subsets analysis, was limited to the second evaluation after radiotherapy. A group of 21 healthy females were used as control. Findings after radiotherapy in asymptomatic patients were compared with findings in a group of patients with radiation pneumonitis. Irradiated patients showed a significantly (p<0.01) greater percentage (29.5+/-15.7%) of BAL lymphocytes than controls (6.2+/-3.3%). No statistical differences existed in BAL findings between the irradiated and unirradiated sides of the chest. Percentages of BAL lymphocytes did not differ significantly between patients who developed subsequent pneumonitis (24.5+/ 13.5%) and those who did not develop pneumonitis (32.8+/-16.5%). Patients with pneumonitis at the time of BAL had significantly higher (p<0.05) alveolar CD4 subset cells (24.8+/-10.2%) than asymptomatic patients (15.2+/-8.9%). Maximal reductions in total lung capacity (p<0.01), and residual volume (p<0.05) occurred 60 days after irradiation. The early lymphocytic alveolitis induced by unilateral thoracic radiotherapy in most patients with breast cancer is always bilateral and does not predict the subsequent development of radiological evidence of pneumonitis. PMID- 10362032 TI - Soluble intercellular adhesion molecule-1 as an early detection marker for radiation pneumonitis. AB - To investigate the role of intercellular adhesion molecule-1 (ICAM-1) in the pathogenesis of radiation pneumonitis and to determine whether the measurement of soluble ICAM-1 (sICAM-1) levels is useful for predicting the onset of pneumonitis, the levels of sICAM-1 were measured in serum and bronchoalveolar lavage (BAL) fluids from patients with lung malignancy who received radiotherapy. A total of 30 patients were irradiated with a total dose of approximately 60 Gy. Blood samples were taken before, midway and after radiotherapy. BAL was also performed before and after radiotherapy in seven cases. The sICAM-1 concentration was measured using an enzyme-linked immunosorbent assay kit with two different monoclonal antibodies. Twelve out of 30 cases developed radiation pneumonitis (pneumonitis group), and the other cases did not (nonpneumonitis group). Serum levels of sICAM-1 after radiotherapy were significantly elevated in the pneumonitis group, but not in the nonpneumonitis group. In some of the cases in the pneumonitis group, sICAM-1 levels began to increase at an early phase of irradiation. In one case of pneumonitis in which BAL was performed, the total cell count and the number of lymphocytes increased markedly, as did the level of sICAM-1 in BAL fluid. These findings suggest that intercellular adhesion molecule 1 may play an important role in the development of radiation pneumonitis and that soluble intercellular adhesion molecule-1 may be a useful marker for the early detection of radiation pneumonitis. PMID- 10362034 TI - Fresh fruit intake and asthma symptoms in young British adults: confounding or effect modification by smoking? AB - Antioxidant vitamins have been postulated as a protective factor in asthma. The associations between the frequency of fresh fruit consumption in summer, and the prevalence of self-reported asthma symptoms were investigated. The analysis was based on 5,582 males and 5,770 females, born in England, Wales and Scotland between March 3-9, 1958 and aged 33 yrs at the time of survey. The 12-month period prevalence of wheeze and frequent wheeze were inversely associated with frequent intakes of fresh fruit and salad/raw vegetables and positively associated with smoking and lower social class. After adjustment for mutual confounding and sex, associations with smoking persisted, but those with social class and salad/raw vegetable consumption lost significance. The frequency of fresh fruit intake was no longer associated with wheeze after adjustment, but was inversely associated with frequent wheeze and speech-limiting attacks. The association with frequent wheeze differed significantly between smoking groups (never, former, current) and appeared to be confined to exsmokers and current smokers. These findings support postulated associations between infrequent fresh fruit consumption and the prevalence of frequent or severe asthma symptoms in adults. Associations appeared to be restricted to smokers, with effect modification as a more likely explanation of this pattern than residual confounding by smoking. PMID- 10362033 TI - Increased nitric oxide in expired air in patients with Sjogren's syndrome. BHR study group. Bronchial hyperresponsiveness. AB - Nitric oxide has an important role in the regulation of airway function and can have pro-inflammatory effects. Bronchial hyperresponsiveness (BHR) and respiratory symptoms are common in patients with Sjogren's syndrome (SS). The aim of this study was to determine whether patients with SS have an increased amount of exhaled NO and whether this NO correlates with respiratory symptoms and BHR. Exhaled NO was measured in 18 patients with SS and 13 normal subjects on three different occasions with intervals of at least 3 days using a chemiluminescence method. Airway responsiveness was assessed with methacholine provocation. Serum levels of myeloperoxidase (MPO), human neutrophil lipocalin (HNL), eosinophil cationic protein (ECP) and eosinophil peroxidase (EPO) were measured. Exhaled NO was significantly higher in patients with SS than in controls (147+/-82 versus 88+/-52 nL x min(-1); mean+/-SD; p=0.041). Exhaled NO was correlated with age (partial r=0.52, p=0.006) and serum HNL (partial r=0.46, p=0.014). There were no significant correlations between exhaled NO and respiratory symptoms, BHR or serum MPO, ECP or EPO. Disease duration was negatively associated with serum MPO (r=-0.47, p=0.043). In patients with SS, a positive correlation was found between symptom score and serum ECP (partial r=0.65, p=0.003) and EPO (partial r=0.62, p=0.004) and a negative correlation with age (partial r=-0.60, p=0.005). In conclusion, elevated levels of exhaled nitric oxide in patients with Sjogren's syndrome were demonstrated. The mechanism underlying this increase in exhaled nitric oxide in Sjogren's syndrome is not known. PMID- 10362035 TI - Asthma symptoms: influence of personality versus clinical status. AB - The hypothesis that symptom-reporting in asthmatics does not necessarily correspond with clinical status, but is related to negative affectivity was investigated. One hundred and sixteen asthmatic patients filled out the Asthma Symptom Checklist (ASC), the Negative Emotionality Scale (NEM), and the McMaster Asthma Quality-of-Life Questionnaire (AQLQ). The patients were grouped as either hospitalized, outpatient previously hospitalized or outpatient not previously hospitalized for asthma. Lung function data and Asthma Severity Scores (ASS) were also collected. The hospitalized group was retested after 3 months. The hospitalized group had lower AQLQ scores, higher ASS scores and worse lung function than both outpatient groups. However, the hospitalized group and the outpatients previously hospitalized group had higher ASC scores and NEM scores than the outpatients not previously hospitalized group. After discharge, when the hospitalized group had become clinically stable, their ASC and NEM scores remained comparable to those of the outpatient previously hospitalized group. Symptom-reporting in asthmatics is not necessarily in accordance with clinical status, but may be directly or indirectly mediated by personality, such as negative affectivity. PMID- 10362036 TI - Exhaled carbon monoxide levels during treatment of acute asthma. AB - Carbon monoxide is known to be present in measurable quantities in the exhaled air of normal subjects and at higher concentrations in asthmatic patients not treated with glucocorticoids. To examine whether exhaled CO is useful in monitoring asthma control, time course changes in peak expiratory flow rate (PEFR) and exhaled CO concentration before and after treatment of acute asthma exacerbations were measured in 20 asthmatic patients. Exhaled CO was measured in triplicate by a portable CO analyser. Exhaled CO was reproducible at all time points. Asthma exacerbations caused a fall in PEFR and a rise in exhaled CO (towards an average of 3.3 parts per million (ppm)) in all patients, and treatment with oral glucocorticoids reversed these changes in both parameters. An improvement of PEFR was closely associated with a reduction of exhaled CO (to an average of 1.5 ppm) after treatment. The maximal exhaled CO concentration significantly correlated with recovery time of PEFR after treatment with oral glucocorticoids (p<0.01). The present study suggests that exhaled CO may be a useful noninvasive means of monitoring the control of asthma. PMID- 10362037 TI - Assessment of bronchial reactivity by forced oscillation admittance avoids the upper airway artefact. AB - The forced oscillation technique (FOT) allows easy assessment of bronchial reactivity. The use of a standard FOT generator (SG) results in changes in respiratory system resistance (delta Rrs,SG) which are affected by an artefact caused by the extrathoracic upper airway (EUA). The aim was to improve the FOT assessment of bronchial reactivity with the SG by computing the change in FOT admittance (delta Ars,SG), which is theoretically unaffected by this artefact. Delta Rrs,SG and delta Ars,SG after bronchial challenge in 17 children were compared with the values measured with a head generator (HG) FOT setup (delta Rrs,HG and delta Ars,HG, respectively), which were taken as a reference, since HG provides data virtually freed from the EUA artefact. At 10 Hz, the SG significantly underestimated the resistance change: delta Rrs,SG=1.77+/-0.62 versus delta Rrs,HG=6.09+/-1.23 hPa x L(-1) x s. Delta Rrs,SG and delta Rrs,HG did not show a significant correlation. By contrast, the amplitude of the change in admittance measured by SG was close to the one obtained with the reference HG: /delta Ars,SG/=29.5+/-4.6 versus /delta Ars,HG/=32.7+/-3.9 mL x hPa(-1) x s(-1). /Delta Ars,SG/ and /delta Ars,HG/ showed a significant correlation (r=0.65, p>0.01). Similar results were found up to 20 Hz. The extrathoracic upper airway artefact was minimized when computing the change in admittance with the standard generator. This forced oscillation technique index may improve the sensitivity in assessing bronchial reactivity with the standard generator setup, which is the most common and easiest to use method for routine lung function testing. PMID- 10362038 TI - Effects of endothelin-1 on airway and parenchymal mechanics in guinea-pigs. AB - The contributions of the airways and the parenchyma to the overall lung mechanical response to endothelin-1 (ET-1) have not been systematically studied. In this investigation, the ET-1 induced changes on lung mechanics in guinea-pigs were separated into airway and parenchymal components. Pulmonary impedance (ZL) data were collected between 0.5 and 21 Hz in six anaesthetized, paralysed, open chest animals by introducing small-amplitude pseudorandom oscillations into the trachea through a wave tube. ZL was calculated before and following intravenous boluses of ET-1, with doses doubled from 0.125-2 microg x kg of body weight(-1). A model containing an airway resistance (Raw) and inertance (Iaw) and tissue damping (G) and elastance (H) was fitted to the ZL spectra in each condition. Parenchymal hysteresis (eta) was calculated as G/H. After each dose, ET-1 induced significant increases in Raw (at peak response mean+/-SEM: 424+/-129%), G (400+/ 80%), H (95+/-22%) and eta (156+/-33%), whereas Iaw decreased following the two highest doses (-291+/-77%). These data suggest that the parenchymal constriction was accompanied by inhomogeneous constriction of the peripheral airways. PMID- 10362039 TI - Role of nitric oxide released from iNANC neurons in airway responsiveness in cats. AB - The precise role of inhibitory nonadrenergic noncholinergic (iNANC) neurons and nitric oxide in airway hyperresponsiveness remains uncertain. The role of NO in the regulation of airway responsiveness was studied in anaesthetized and mechanically ventilated cats. To assess airway responsiveness, the changes in total pulmonary resistance (RL) produced by delivering serotonin aerosol to the airways were measured before and after N(omega)-nitro-L-arginine methyl ester (L NAME), or a ganglionic blocker, hexamethonium, which has been reported to block iNANC. Serotonin was chosen because it causes bronchoconstriction in part by neural reflex. To further clarify the mechanism(s) involved, the effect of inhaled capsaicin was also determined in animals with sustained bronchoconstriction induced by serotonin after treatment with atropine and propranolol. Inhibition of NO synthase by L-NAME or blockade of iNANC neurons by hexamethonium significantly increased airway responsiveness. However, addition of L-NAME did not further increase airway responsiveness in animals treated with hexamethonium. In the presence of atropine and propranolol, inhaled capsaicin caused a marked bronchodilation during serotonin-induced sustained bronchoconstriction. The bronchodilation induced by capsaicin was significantly suppressed by hexamethonium and by L-NAME. These results suggest that the nitric oxide released from inhibitory nonadrenergic noncholinergic neurons is important in modulating the airway responsiveness of cats in vivo. PMID- 10362040 TI - Subjective differentiation of normal and pathological bronchi on thin-section CT: impact of observer training. AB - The effect of observer training on sensitivity, specificity and interobserver agreement in the differentiation between normal and pathological bronchi on computed tomography (CT) was studied. The wall thickness of bronchi with normal walls and with pathologically thickened walls were subjectively scored by three independent observers before and after a training period of 2 weeks. Sensitivity, specificity and interobserver agreement were calculated for reading sessions before and after training. Increase and decrease in agreement after training were determined. There was a statistically significant difference (p=0.001) between objectively measured wall thickness of normal and pathological bronchi, both for reference bronchi and for bronchi used for reading sessions. While training increased interobserver agreement, it had no effect on sensitivity (0.46 versus 0.44 after training) and specificity (0.71 versus 0.72 after training) in detecting pathological bronchi. Increased agreement after training was significantly (p=0.001) more frequent than decreased agreement. There is a discrepancy between the effect of training on interobserver agreement and on sensitivity and specificity in the subjective differentiation between normal and pathological bronchi. Interobserver agreement alone is not a reliable indicator of a beneficial effect of training in the evaluation of this parameter. PMID- 10362041 TI - Variability of aerosol delivery via spacer devices in young asthmatic children in daily life. AB - Pressurized metered dose inhalers (pMDI) are widely used together with spacers for the treatment of asthma in children. However, the variability of daily medication dose for pMDI/spacer combinations is not known. Electrostatic charge is a potential source of dose variability. Metal spacers have no static charge. This study assessed and compared within-subject variability of aerosol delivery of metal and plastic spacers. This was a randomized, crossover study in children with stable asthma aged 1-4 (group I, n=17) and 5-8 (group II, n=16) yrs. In both groups the amount of drug delivered to the mouth by a metal spacer (Nebuchamber) and one of two plastic (polycarbonate) spacers, i.e. Babyhaler in group I and Volumatic in group II was measured. The metal and plastic spacers were tested at home in a randomized order for 7 days each, using budesonide (200 microg b.i.d.). Aerosol was collected on a filter positioned between spacer and facemask or mouth. Budesonide on the filter was assessed by high performance liquid chromatography. The mean filter dose for each child (mean+/-SD) during the 7 days was expressed as a percentage of the nominal dose. Within-subject variability was expressed as coefficient of variation (CV). Mean filter dose in group I was 41.7+/-10.1% for Nebuchamber and 26.0+/-4.0% for Babyhaler (p<0.001). Mean filter dose in group II was 50.2+/-9.2% for Nebuchamber and 19.4+/-7.2% for Volumatic (p<0.001). Mean CV in group I was 34% for Nebuchamber and 37% for Babyhaler (p=0.44). Mean CV in group II was 23% for Nebuchamber and 34% for Volumatic (p=0.003). There was substantial within-subject dose variability in aerosol delivery in children using a pMDI/spacer at home. This variability was lower for the metal than for the plastic spacer in children 5-8 yrs of age. The dose delivered to the mouth was about two-fold higher for the metal than the plastic spacer independent of age. PMID- 10362042 TI - Measurement of airway resistance using the interrupter technique in preschool children in the ambulatory setting. AB - This study describes the feasibility, repeatability, and interrater reliability of the measurement of airway resistance by the interrupter technique (Rint) in children 2-5 yrs of age, and examines whether reversibility to bronchodilator can be demonstrated in wheezy children. The mean of six Rint values was taken as a measurement. If subjects could complete one measurement and then a second 15 min after bronchodilator, baseline testing and reversibility testing were considered feasible. To measure repeatability, two measurements 30 s apart and measurements before and 15 min after placebo bronchodilator were compared. Measurements by two testers were compared for interrater reliability. Change in Rint in wheezy children was measured after bronchodilator. Fifty-six per cent of 2-3-yr-olds (n=79), 81% of 3-4-yr-olds (n=104) and 95% of 4-5-yr-olds (n=88) completed baseline testing, and 53%, 71% and 91% completed reversibility testing. Baseline measurements were 0.47-2.56 kPa x L(-1) x s. Repeatabilities (2 SD of the mean differences between measurements) at 30 s in the three age bands were 0.21, 0.17 and 0.15 kPa x L(-1) x s and 0.19 kPa x L(-1) x s after placebo. Using 0.21 kPa x L(-1) x s as the threshold for reversibility, reversibility was demonstrated in most wheezy children. Interrater reliability was 0.15 kPa x L(-1) x s. Preschool children can undertake measurements of airway resistance by the interrupter technique in ambulatory settings and reversibility to bronchodilator in wheezy children can be demonstrated. This technique promises to be a useful clinical and research tool. PMID- 10362043 TI - Differences in nasal cellular infiltrates between allergic children and age matched controls. AB - Little is known about the cellular infiltrates in the nasal mucosa of children. This study was set up to compare the nasal cellular infiltrates in biopsy specimens from allergic children and controls. Atopic children were distinguished from controls on the basis of symptoms of allergic rhinitis and/or asthma, total serum immunoglobulin (Ig)E, family history and specific serum IgE to food and aeroallergens. Fifteen allergic patients (median age 4.3 yrs) and 15 age-matched nonallergic control subjects were evaluated. The number of cells positive for CD1a, CD4, CD8, CD19, CD68, chymase, tryptase, IgE and major basic protein was determined in the mucosa of the inferior turbinate. A significantly higher number of IgE-positive cells and mast cells was found in the epithelia of the allergic group. In the lamina propria, higher numbers of IgE-positive cells and eosinophils were found. Langerhans' cells positive for IgE were only seen in allergic children with specific serum IgE against aeroallergens. These children also had a higher number of IgE-positive mast cells compared to controls and atopic children without specific serum IgE. These results show that the nasal cellular infiltrates of allergic children differ from nonallergic control subjects. Prior to the detection of specific serum immunoglobulin E, cellular changes can be found in the nasal mucosa of atopic children. PMID- 10362044 TI - Energy balance during acute respiratory exacerbations in children with cystic fibrosis. AB - Acute respiratory exacerbations have been proposed to contribute to the negative energy balance which causes undernutrition in cystic fibrosis. However, no studies have measured their effect on all components of energy balance. The aim of this study was to measure the effect of an acute respiratory exacerbation on energy balance. Fourteen children (six females, eight males, mean+/-SD age 9.9+/ 2.4 yrs) were studied when well and during the course of an acute respiratory exacerbation treated with intravenous antimicrobial therapy. The total energy expenditure was measured using the doubly-labelled water method, resting energy expenditure by ventilated hood indirect calorimetry, energy intake by household measures records, and fat malabsorption from measurements of dietary fat intake and faecal fat output. The exacerbation was associated with a significant reduction in energy intake (mean paired difference 47 kJ x kg of body weight(-1) x day(-1), p<0.01). Changes in fat malabsorption and resting energy expenditure were negligible. The absence of significant changes in body weight and composition, together with the trend towards lower total energy expenditure, suggested no marked negative energy balance during the exacerbation. In conclusion, treatment of acute respiratory exacerbation with intravenous antimicrobial therapy represents a relatively minor challenge to energy balance and nutritional status in children with cystic fibrosis. PMID- 10362045 TI - Soluble intercellular adhesion molecule-1 in the bronchoalveolar lavage fluid of normal children exposed to parental cigarette smoke. AB - This study sought to test the hypothesis that normal children exposed to parental cigarette smoke have increased bronchoalveolar lavage (BAL) fluid levels of soluble intercellular adhesion molecule (sICAM)-1. Cells and solutes from the lower airway of normal children were obtained by nonbronchoscopic BAL using three aliquots of 1 mL x kg body weight(-1) normal saline, prior to elective orthopaedic surgery. Children with evidence of recent or ongoing infection, atopic disease, previous history of wheeze, and chronic respiratory symptoms were excluded. Twelve children with parents who smoked (group 1) were paired with 12 age- and weight-matched controls with self-reported nonsmoking parents (group 2). There was no significant difference (group 1 versus 2) in the volume of BAL fluid recovered (median 29.0 versus 28.7 mL), the percentage of alveolar macrophages (92.5 versus 91.8%), neutrophils (1.1 versus 2.1%), lymphocytes (5.3 versus 5.6%) and eosinophils (0 versus 0%), and the total BAL fluid leukocyte concentration (80 versus 61 x 10(3) cells x mL(-1)). BAL fluid albumin concentration was similar between the two groups (0.033 versus 0.020 mg x mL(-1)). sICAM-1 was detected in all BAL fluid samples, and was significantly increased in group 1 (39.2 versus 22.5 ng x mL(-1), p<0.01). It was concluded that exposure of children to parental cigarette smoke is associated with increased soluble intercellular adhesion molecule-1 concentrations in the bronchoalveolar lavage fluid and this may reflect an altered activation of pulmonary immune cells. PMID- 10362046 TI - Reduced expression of the alphabeta T-cell antigen receptor by alveolar T-cells. AB - A previous study revealed that reduced expression (modulation) of the CD3 antigen is a common characteristic of alveolar T-cells in health and disease. As CD3 molecules are noncovalently bound to T-cell antigen receptors (TCR), it was hypothesized that modulation of TCR was also a feature of alveolar T-cells. To demonstrate this, lymphocytes from bronchoalveolar lavage fluid were stained with an anti-alphabeta TCR antibody and analysed by flow cytometry. The expression of alphabeta TCR by alveolar T-cells was evaluated by calculating mean fluorescence intensity (MFI) and was compared with alphabeta TCR expression by autologous blood T-cells. As anticipated from a previous study, modulation of TCR was observed not only in healthy volunteers but also in patients with pulmonary sarcoidosis, other pulmonary diseases, and nonpulmonary diseases. There were no significant differences in MFI of alveolar T-cells among the study groups. The degree of modulation assessed by the difference of MFI between blood and alveolar T-cells was greater for CD4+ cells than for CD8+ cells owing to the higher MFI of CD4+ blood T-cells. Coculture of alveolar macrophages with blood T-cells in vitro induced partial modulation of TCR. These results demonstrate the ubiquity of modulation of T-cell receptors on alveolar T-cells and suggest, in contrast to a previous report by other investigators that it is caused by some nonantigenic mechanism possibly inherent in the alveolar milieu. The implications of this phenomenon in in vivo immune responses of the lung need to be examined. PMID- 10362047 TI - Monocyte chemotactic factors released from type II pneumocyte-like cells in response to TNF-alpha and IL-1alpha. AB - It has been reported that tumour necrosis factor (TNF)-alpha and interleukin (IL) 1 induce the release of monocyte chemotactic factors (MCF), including chemokines, from A549 cells, an alveolar type II cell line. However, the relative contribution of these chemokines to MCF is still uncertain. In the present study, the relative contribution of various chemokines released from A549 cells acting as MCF upon stimulation by TNF-alpha and IL-1alpha, was evaluated. TNF-alpha and IL-1alpha induced the release of MCF in a dose- and time-dependent manner (p<0.001). The release of MCF was inhibited by cycloheximide and lipoxygenase inhibitors. Molecular sieve column chromatography revealed multiple peaks of MCF (near 60 kDa, 25-22 kDa, 15-13 kDa, 8 kDa, and 400 Da). Leukotriene B4 (LTB4) receptor-antagonists inhibited MCF by 50% after 24 h and 30% after 72 h. Monocyte chemoattractant protein-1 (MCP-1), transforming growth factor (TGF)-beta, "regulated on activation, normal T-cells, expressed and secreted" (RANTES), and granulocyte-macrophage colony- stimulating factor (GM-CSF) were released significantly in response to IL-1alpha and TNF-alpha, and antibodies to MCP-1, GM CSF, and RANTES inhibited MCF activity by 40, 5 and 20% after 24 h, and by 50, 20, and 10% after 72 h, respectively. Each antibody or LTB4 receptor-antagonist inhibited the corresponding column chromatography-separated molecular weight peak of MCF. These data suggest that A549 cells release monocyte chemoattractant protein-1 as the predominant monocyte chemotactic factor rather than granulocyte macrophage colony-stimulating factor, RANTES, and transforming growth factor beta, and that leukotriene B4 is constitutively released as a monocyte chemotactic factor. PMID- 10362048 TI - N-isobutyrylcysteine, a donor of systemic thiols, does not reduce the exacerbation rate in chronic bronchitis. AB - N-isobutyrylcysteine (NIC), a new thiol compound that is not rapidly hydrolysed to give higher levels of free thiols in the body than N-acetylcysteine (NAC), was used to test if the effect of NAC on exacerbations in chronic bronchitis was an effect of the unhydrolysed thiol compound. Smokers or exsmokers with chronic bronchitis forced expiratory volume in one second (FEV1) >40% and reversibility < or = 10% predicted were treated with oral NIC 300 mg b.i.d. or placebo for 24 weeks. Steroids, NAC, antibiotics, and nonsteroid anti-inflammatory drugs use were restricted. Exacerbations were recorded by a respiratory symptom diary card and the time to onset of the first exacerbation after the start of treatment was measured using life-table analysis. Spirometry was performed at each visit. Six hundred and thirty-seven patients were randomized to treatment with NIC (n=316) or placebo (n=321). NIC did not prolong the time to first exacerbation (life table analysis, p=0.59) and no increase in FEV1 or forced vital capacity was observed. Altered taste perception, taste loss and anosmia occurred more often in the NIC group (p<0.001). In conclusion, N-isobutyrylcysteine, a N-acetylcysteine like drug with a greater bioavailability has, contrary to N-acetylcysteine, no effect on exacerbations in chronic bronchitis. This suggests that the effect of N acetylcysteine on exacerbations in chronic bronchitis is not due to the relatively low free thiol levels (other than glutathione) produced by N acetylcysteine therapy. PMID- 10362049 TI - Antibiotic use in patients admitted with acute exacerbations of chronic obstructive pulmonary disease. AB - The objective of this report was to document the pattern of initial antibiotic prescribing in acute exacerbations of chronic obstructive pulmonary disease (COPD) in a hospital setting. All episodes of acute exacerbation of COPD, as diagnosed by the admitting doctor, in one hospital in the period January to May 1996, were identified. Case notes were reviewed retrospectively. Cases of radiographic pneumonia, bronchiectasis and incorrectly coded admissions were excluded. Symptoms, microbial cultures and initial antibiotic therapies were recorded. One hundred and fifty-nine patient episodes were identified; 40 were excluded yielding a sample of 119. Nineteen case notes were unavailable leaving a sample of 100 (84%) episodes. Eighty were treated with antibiotics on admission; amoxycillin was the most frequently prescribed, in 46 (58%) episodes. Of the antibiotic treated group, 42 (53%) patients were given dual therapy, most commonly a macrolide antibiotic with either amoxycillin or a cephalosporin. Intravenous treatment was used in 22 (28%) cases. The duration of intravenous treatment was >48 h in 12 (15%) cases. A total of 76 sputum samples were analysed from 55 patient episodes: 34 (45%) were culture positive. In 15 (27%) patient episodes, antibiotic therapy was changed or instituted on the basis of culture results. These data suggest that antibiotic treatment is not optimal, with overuse of antibiotics, especially intravenous and dual therapy. PMID- 10362050 TI - The cellular composition of induced sputum in chronic obstructive pulmonary disease. AB - Asthma and chronic obstructive pulmonary disease are characterized by airway inflammation, which can be assessed by bronchoscopic techniques as well as by the analysis of induced sputum. A method to induce sputum with inhaled hypertonic saline was adapted for use in 21 chronic obstructive pulmonary disease (COPD) patients (mean baseline forced expiratory volume in one second (FEV1) 1.60 L, or 54% predicted) and in 16 healthy volunteers. The success rate and safety of the method, were investigated along with the reproducibility of cell counts and differences in cell counts between both groups. All subjects produced adequate samples and the procedure did not alter spirometric values. A marked sputum neutrophilia was noted in patients with COPD (74.9+/-4.7%), whereas mainly macrophages were seen in healthy volunteers (74.0+/-4.0%). Reliability of the cell counts was high, both within investigators (r=0.99 neutrophils, r=0.99 macrophages) and between investigators (r=0.95 neutrophils, r=0.77 macrophages). In patients with COPD, an inverse correlation was noted between percentage of neutrophils and FEV1 (r(s)=-0.48, p<0.05). Immunostaining revealed a large proportion of activated macrophages in both groups. It was concluded that induction of sputum is a safe and reproducible method to study the composition of airway secretions in patients with chronic obstructive pulmonary disease. PMID- 10362051 TI - Effect of temperature on lung function and symptoms in chronic obstructive pulmonary disease. AB - The present study investigated whether falls in environmental temperature increase morbidity from chronic obstructive pulmonary disease (COPD). Daily lung function and symptom data were collected over 12 months from 76 COPD patients living in East London and related to outdoor and bedroom temperature. Questionnaires were administered which asked primarily about the nature of night time heating. A fall in outdoor or bedroom temperature was associated with increased frequency of exacerbation, and decline in lung function, irrespective of whether periods of exacerbation were excluded. Forced expiratory volume in one second (FEV1) and forced vital capacity (FVC) fell markedly by a median of 45 mL (95% percentile range: -113-229 mL) and 74 mL (-454-991 mL), respectively, between the warmest and coolest week of the study. The questionnaire revealed that 10% had bedrooms <13 degrees C for 25% of the year, possibly because only 21% heated their bedrooms and 48% kept their windows open in November. Temperature-related reduction in lung function, and increase in exacerbations may contribute to the high level of cold-related morbidity from chronic obstructive pulmonary disease. PMID- 10362052 TI - Altered expression of myosin heavy chain in the vastus lateralis muscle in patients with COPD. AB - This study was designed to further characterize peripheral skeletal muscle alterations in patients with chronic obstructive pulmonary disease (COPD) and to evaluate the possible relationship between myosin heavy chain (MyoHC) isoform expression and exercise tolerance in these individuals. MyoHC composition from biopsy of the vastus lateralis muscle was examined in 12 COPD patients (forced expiratory volume in one second (FEV1)=31+/-9% predicted, peak oxygen consumption (V'O2)=15+/-4 mL x kg(-1) x min(-1)) and 10 age-matched normal male subjects (peak V'O2=20+/-5 mL x kg(-1) x min(-1)). The proportion of MyoHC type I was smaller in COPD than in normals (27+/-17% versus 41+/-9%, p<0.05) with an increase in MyoHC type IIa (51+/-15% versus 39+/-9%, p<0.05) and the proportion of MyoHC type IIx being comparable between both groups. A significant relationship was found between peak V'Oo2 mL x kg(-1) x min(-1) and FEV1 % pred (r=0.91, p<0.0001) and the percentage of MyoHC type I (r=0.61, p=0.016). In stepwise multiple regression, only FEV1 % pred was found to be a significant determinant of peak V'O2 (p<0.0001). This variable explained 83% of the total variance of peak V'O2. In summary, this study showed considerable modifications in the phenotypic expression of the myosin heavy chain in the vastus lateralis muscle in patients with chronic obstructive pulmonary disease. An independent effect of myosin heavy chain expression on exercise capacity was not found. These results suggest that chronic inactivity and muscle deconditioning may not be the sole factors explaining peripheral muscle dysfunction in patients with chronic obstructive pulmonary disease. PMID- 10362053 TI - Respiratory rehabilitation in chronic obstructive pulmonary disease: predictors of nonadherence. AB - Rehabilitation is now an integral part of chronic obstructive pulmonary disease (COPD) management. The objective of the study was to determine predictors of nonadherence to a COPD rehabilitation programme. Patients attending a COPD clinic were invited to participate in a 4 week, hospital-based, outpatient, COPD rehabilitation programme conducted predominantly by respiratory physiotherapists. All potential participants undertook an interviewer administered questionnaire addressing social, economic, psychological and healthcare factors, and underwent baseline physiological measures. Subsequently they were classified as: 1) "adherent" group who completed the total programme (n=55) or 2) "nonadherent" group who refused or began but did not complete the programme (n=36). The nonadherent group compared to the adherent group were more likely to be divorced (22 versus 2%, p<0.005), live alone (39 versus 14%, p<0.02), and to live in rented accommodation (31 versus 6%, p<0.005). There were no differences between the two groups in terms of baseline physiological parameters (forced expiratory volume in one second, forced vital capacity, 6-min walk distance, oxygen saturation, perceived dyspnoea), quality of life domains (Chronic Respiratory Disease Questionnaire), or indices of COPD-related morbidity. The nonadherent group were more likely to be current smokers (28 versus 8%, p<0.02) and less likely to use inhaled corticosteroids (16 versus 42%, p<0.005). The nonadherent group was not significantly likely to be depressed, anxious, prone to hyperventilation or to have had previous emotional counselling and was more likely to be dissatisfied with disease-specific social support (51 versus 2%, p<0.001). In conclusion, a substantial proportion of eligible subjects who did not participate in a chronic obstructive pulmonary disease rehabilitation programme were not more physiologically impaired, but were more likely to be: socially isolated, lack chronic obstructive pulmonary disease-related social support, still be smoking and be less compliant with other healthcare activities. Identification of one or more of these factors reliably allows prediction for nonadherence to a rehabilitation programme. PMID- 10362055 TI - Changes in airway resistance induced by nasal or oral intermittent positive pressure ventilation in normal individuals. AB - Nasal intermittent positive-pressure ventilation (nIPPV) is used for the treatment of respiratory failure in patients with neuromuscular disease. The aim of the present study was to demonstrate that nIPPV may activate nose receptors, the consequence of which being reflex changes in lung resistance. The changes in interrupter resistances (Rint) in response to nIPPV were tested before and after local anaesthesia of the nasal mucosa in normal subjects. They were compared to the Rint changes induced by oral intermittent positive-pressure ventilation (oIPPV) in the same individuals. Rint was measured during 10-min periods of nIPPV or oIPPV at a constant rate (15 L x min(-1)), but at two different stroke volumes (0.8 and 1.2 L). Inspired temperature and relative humidity were held constant. nIPPV with 1.2 L (17 mL x kg(-1)) significantly increased the Rint value (+22%). This effect disappeared after nose anaesthesia or after inhalation of a cholinergic antagonist. oIPPV never changed Rint, even though the associated hypocapnia was present and more accentuated than during nIPPV. Adding CO2 to the inspired gas during nIPPV and oIPPV trials suppressed the Rint changes. The present study suggests the existence of a nasopulmonary bronchoconstrictor reflex elicited through the stimulation of nasal mechanoreceptors, their activity being markedly influenced by the changes in expired CO2 concentration. PMID- 10362056 TI - Effects of chemical feedback on respiratory motor and ventilatory output during different modes of assisted mechanical ventilation. AB - The purpose of the study was to examine the effects of chemical feedback on respiratory motor and ventilatory output in conscious subjects ventilated on various modes of assisted mechanical ventilation. Seven subjects were connected to a ventilator and randomly ventilated on assist-volume control (AVC), pressure support (PS) or proportional assist ventilation (PAV). On each mode, the assist level was set to the highest comfortable level. Airway and oesophageal (Poes) pressures, tidal volume, respiratory frequency (fR) and end-tidal carbon dioxide tension (PET,CO2) were measured breath-by-breath. When the subjects were stable on each mode, the fraction of inspired carbon dioxide (FI,CO2) was increased stepwise, and changes in minute ventilation (V'E) and respiratory motor output, estimated by the pressure-time product of all the respiratory muscles per breath (PTPrm) and per minute (PTPminute), were observed. At zero FI,CO2, PTPminute/PET,CO2 did not differ between modes, while V'E/ PTPminute was significantly lower with PAV than that with PS and AVC. As a result V'E/PET,CO2 was significantly lower with PAV, preventing, unlike AVC and PS, a significant drop in PET,CO2. With PAV, independent of CO2, V'E/PTPminute remained constant, while it decreased significantly with increasing CO2 stimulus with PS and AVC. At high PET,CO2 respiratory effort was significantly lower with PAV than that with PS and AVC. In conclusion, the mode of mechanical ventilation modifies the effects of chemical feedback on respiratory motor and ventilatory output. At all carbon dioxide stimulus levels neuroventilatory coupling was better preserved with proportional assist ventilation than with pressure support and assist-volume control ventilation. PMID- 10362054 TI - Influence of immersion on respiratory requirements during 30-min cycling exercise. AB - Immersion is considered to facilitate exercise-based rehabilitation. However, the drag effect of moving limbs in water, likely to increase the respiratory requirements at exercise, is not mentioned in many reports. The energetic and ventilatory requirements of 30 min steady state cycling exercise performed by healthy male subjects in air and during immersion up to the xiphoid in 33 degrees C water were compared. In the first experimental series nine subjects exercised at the same 60% maximal oxygen consumption (V'O2,max) in air and water. In the two ambient conditions, ventilatory variables had similar values, but the ergometric setting had to be reduced during water immersion so that the workload rated only 69+/-20 W (mean+/-SD) in water versus 121+/-32 W (p<0.001) in air. In the second experimental series, the same ergometric work load (122 W) was achieved by nine subjects with an average V'O2 of 2,210+/-300 mL x min(-1) in air versus 2,868+/-268 mL x min(-1) in water (p<0.001). Resting water immersion caused a marked trend for decreasing vital capacity (p=0.06), but no modification of other ventilatory variables. During exercise at similar V'O2, the average values of minute ventilation (V'E), tidal volume (VT), respiratory frequency (fR), tidal inspiratory time (VT/tI) were not different between water and air. However, at similar ergometric workload, V'E, VT, fR, VT/tI and plasma lactate levels were significantly higher in water than in air. Such consequences of the drag effect of water upon limb movements have not been reported in previous studies relying on shorter exercise bouts. Thus, maintaining steady exercise levels in water either led to a decrease in the workload or required a 25% higher oxygen consumption than in air. These findings may be relevant to the prescription of water immersion rehabilitation programmes. PMID- 10362057 TI - Severe respiratory failure requiring ICU admission in bone marrow transplant recipients. AB - Two groups of bone marrow transplant (BMT) recipients with febrile noncardiogenic respiratory failure requiring intensive care unit (ICU) admission, in the early phase of bone marrow transplantation were compared: those who had proven infectious pneumonia and those in whom bronchoalveolar lavage (BAL) failed to establish a diagnosis. Thirty-eight consecutive neutropenic BMT recipients admitted to an ICU with febrile noncardiogenic respiratory failure were enrolled. All of them underwent a BAL with viral, fungal, bacterial, and histopathological examinations. Lung biopsies were performed in nonsurviving patients in order to compare with BAL results. Haematological, biological, respiratory failure and other organ failure parameters, infectious results, outcome, and lung biopsy results were evaluated. BAL allowed an infectious diagnosis to be established in 16 BMT recipients. No aetiology was proven in 22 patients. Without a significant difference in respiratory failure parameters on ICU admission, noninvasive continuous positive airway pressure ventilation, which was given to 11 patients in each group, was significantly more successful in patients with proven infectious pneumonia (6 of 11 versus 0 of 11 patients) and enabled endotracheal intubation to be avoided in significantly more patients with infectious disease (10 of 16 versus 22 of 22 patients). The evolution of patients without diagnosis was significantly different with more frequent renal failure, hepatic failure, and death (20 of 22 versus 9 of 16 patients). Post mortem biopsies confirmed the absence of micro-organisms, but endothelial damage and fibrosis was found in 14 of the 22 patients. In conclusion, in the early phase of bone marrow transplantation the recipients without proven aetiology of pneumonia have a worse outcome than grafted patients with proven infectious pneumonia. PMID- 10362058 TI - Multidisciplinary approach to management of postintubation tracheal stenoses. AB - The optimal management of postintubation tracheal stenosis is not well defined. A therapeutic algorithm was designed by thoracic surgeons, ear, nose and throat (ENT) surgeons, anaesthetists and pulmonologists. Rigid bronchoscopy with neodymium-yttrium aluminium garnet (Nd-YAG) laser resection or stent implantation (removable stent) was proposed as first-line treatment, depending on the type of stenosis (web-like versus complex stenosis). In patients with web-like stenoses, sleeve resection was proposed when laser treatment (up to three sessions) failed. In patients with complex stenoses, operability was assessed 6 months after stent implantation. If the patient was judged operable, the stent was removed and the patient underwent surgery if the stenosis recurred. This algorithm was validated prospectively in a series of 32 consecutive patients. Three patients died from severe coexistent illness shortly after the first bronchoscopy. Of the 15 patients with web-like stenosis, laser resection was curative in 10 (66%). Among the 17 patients with complex stenoses, three remained symptom-free after stent removal. Bronchoscopy alone was thus curative in more than one-third of the patients. Six patients underwent surgery, two after failure of laser resection and four after failure of temporary stenting. Surgery was always performed with the patient in good operative condition. Palliative stenting was the definitive treatment in nine cases. Tracheostomy was the definitive solution in two cases. This approach, including an initial conservative treatment, depending on the type of the stenosis, appears to be applicable to almost all patients and allows secondary surgery to be performed with the patient in good condition. PMID- 10362059 TI - Two-colour flow-cytometric analysis of pulmonary alveolar macrophages from smokers. AB - The study of alveolar macrophages (AM) from smokers by flow cytometry (FCM) has been limited by strong autofluorescence and the lack of reliable markers to identify macrophages. Crystal violet quenching was reported to be effective in reducing autofluorescence of AM. CD68 is a marker for macrophages in immunohistochemistry, but has been less useful in FCM because of poor surface expression. This study evaluated the effectiveness of a method for two-colour FCM analysis of AM combined with membrane permeabilization and crystal violet quenching. Bronchoalveolar lavage cells, fixed in 4% paraformaldehyde and permeabilized using 0.5% Triton X100, were incubated with fluorescent-labelled antibodies for 30 min and quenched with a saturated crystal violet solution. Two colour FCM was then performed using forward/side scatter gating to select AM. Autofluorescence at 525 nm (fluorescein isothiocyanate) and 575 nm (phycoerythrin) markedly decreased after quenching. After permeabilization, 97.1+/-2.8% of the gated cells were CD68+, while 53.9+/-18.6% of the AM were positive without permeabilization. CD68+ cells were sorted and proved to be AM morphologically. Analysis of CD71 (transferrin receptor) expression by FCM correlated with immunocytochemistry (r=0.77, p<0.05). The permeabilization/quenching technique, therefore, represents a satisfactory means to evaluate alveolar macrophages by flow cytometry. PMID- 10362061 TI - Outcome of asthma: longitudinal changes in lung function. AB - Current knowledge about factors determining outcome of asthma is limited, but observations over the last few decades suggest that active asthma has a negative impact on the longitudinal changes in lung function. This review aims to give an overview of the present knowledge concerning longitudinal changes in lung function, including clinical markers for distinctly poor outcome with regard to lung function, in children and adults suffering from asthma. The majority of patients with asthma have a good prognosis. However, some patients with asthma, especially those with more severe disease, are at risk of impaired growth of lung function during childhood, a lower maximally attained level of lung function and excessive decline in lung function in adulthood, which may lead to life threatening lung function impairment. Clinical markers of poorly controlled airway inflammation appear to have a negative impact on the longitudinal changes in lung function, and disease progression to nonreversible airflow obstruction may be observed in a minority of patients with asthma. Early intervention with anti-inflammatory therapy may improve the short-term outcome of asthma, but long term controlled studies are clearly needed in order to verify whether or not treatment, especially with inhaled corticosteroids, according to the current international guidelines alters the natural history of asthma, i.e. disease progression with regard to changes in lung function and possible development of nonreversible airflow obstruction. PMID- 10362060 TI - Prospective multicentre study on the evaluation of antituberculosis treatment results in Italy: comparison of the culture- versus the smear-based methods. National AIPO Tuberculosis Study Group. AB - Cohort analysis of treatment outcomes is the most informative technique to evaluate the tuberculosis (TB) control programme. The aim of the study was to assess treatment outcomes comparing the smear- versus the culture-based methods, using data on TB patients treated under programme conditions in Italy. This was a prospective monitoring study based on the standardized collection of forms from a representative sample of Italian TB Units. The forms, with individual data, were reviewed and analysed on a quarterly basis according to the principles of cohort analysis, using both the smear- and culture-based methods. The complete bacteriological profile of patients was analysed at diagnosis and at completion of treatment. Nine hundred and ninety-two TB cases were notified. Among 681 pulmonary cases, 368 cases were culture-confirmed at diagnosis (333 new and 35 retreatment cases, 293 being sputum smear positive, 79.6%). At the end of treatment, out of the 333 new culture-confirmed cases, 136 (40.8%) were defined "cured" using the culture-based method and 108 (32.4%) using the smear-based method (p<0.05, chi2 test). The culture-based method is the recommended tool to evaluate pulmonary tuberculosis treatment results. Culture allows a more precise definition of a "cured" patient in both sputum smear positive and negative tuberculosis cases. PMID- 10362062 TI - Surgical aspects and techniques of lung volume reduction surgery for severe emphysema. AB - Lung volume reduction surgery (LVRS) has become an accepted procedure for palliative treatment of diffuse, nonbullous emphysema. Single or multiple peripheral segmental wedge resections of the most destroyed areas of the lungs are performed with the use of stapling devices, in order to decrease hyperinflation and restore diaphragmatic function. Median sternotomy, videoendoscopy or anterior muscle sparing thoracotomies have been used as surgical approaches. The functional improvement after bilateral resections exceed those after a unilateral approach. LVRS has demonstrated its potential as an alternative to transplantation, and with growing experience, the indications for the procedure have been widened. In selected patients with peripheral lung cancer who have been considered unsuitable for a surgical resection, the combination of both tumour resection and LVRS has successfully been performed. In contrast to LVRS, laser surgery of the emphysematous lung has been abandoned in most institutions. PMID- 10362064 TI - Systemic lupus erythematosus, eosinophilia and Loffler's endocarditis. An unusual association. AB - A 24-yr-old male, known since the age of 11 to have a nonerosive arthritis and later diagnosed as having systemic lupus erythematosus (SLE), developed subacute heart failure with diffuse lung infiltrates and died suddenly after having presented a moderate hypereosinophilia for 6 months for which no other causes besides the SLE were found. A post mortem examination revealed Loffler's endocarditis (endocarditis parietalis fibroplastica) with acute pulmonary capillaritis. This represents Loffler's endocarditis in the setting of SLE. To the best of the authors' knowledge, this association has not been reported before. PMID- 10362063 TI - Bronchiolitis obliterans organizing pneumonia associated with polymyalgia rheumatica. AB - The association of bronchiolitis obliterans organizing pneumonia (BOOP) with polymyalgia rheumatica is rare, and only one case has previously been described. This study reports on the case of an 80 yr-old male who presented with malaise, nonproductive cough and exertional dyspnoea for several weeks, along with a history of bilateral shoulder and pelvic girdle pain of several months' duration. The chest radiograph revealed a pneumonic infiltrate in the right lower lobe, which was unresponsive to antibiotics. Bronchoscopy, bronchoalveolar lavage and a transbronchial lung biopsy established the diagnosis of BOOP. The patient improved consistently on steroids. As in other connective diseases, organizing pneumonia may be one of the early manifestations of polymyalgia rheumatica. PMID- 10362065 TI - Constrictive bronchiolitis obliterans and paraneoplastic pemphigus. AB - Constrictive bronchiolitis obliterans is rare, and the pathogenesis of the disease often remains unknown. This study reports on the case of a 38 yr-old female with constrictive bronchiolitis obliterans and paraneoplastic pemphigus associated with malignant lymphoma. The patient developed progressive obstructive lung disease. The chest radiograph showed almost normal lungs. Paraneoplastic pemphigus is a newly described syndrome in which patients have autoantibodies binding to some epithelia, including in the respiratory tract. The disease develops in association with non-Hodgkin's lymphomas or other malignant neoplasms. The case presented here suggests that constrictive bronchiolitis obliterans associated with paraneoplastic pemphigus may be one of the facets of autoimmune responses in this context. PMID- 10362066 TI - Plasma exchange: an option for the treatment of life-threatening status asthmaticus in pregnancy. PMID- 10362067 TI - Five-year follow-up of Algerian victims of the "Ardystil syndrome". PMID- 10362068 TI - In memory of the late Professor Noburo Kamiya. PMID- 10362069 TI - A 60 kDa plasma membrane protein changes its localization to autophagosome and autolysosome membranes during induction of autophagy in rat hepatoma cell line, H 4-II-E cells. AB - We previously reported the preparation and characterization of an antibody against membrane fraction of autolysosomes from rat liver (J. Histochem. Cytochem. 38, 1571-1581, 1990). Immunoblot analyses of total membrane fraction of a rat hepatoma cell line, H-4-II-E cells by this antibody suggested that H-4-II-E cells expressed several autolysosomal proteins, including a protein with apparent molecular weight of 60 kDa. It was suggested that this 60 kDa protein was a peripheral membrane protein, because it was eluted from the membrane by sodium carbonate treatment. We prepared an antibody against this 60 kDa protein by affinity purification method, and examined its behavior during induction of autophagy. Autophagy was induced by transferring the cells from Dulbecco's modified Eagle medium (DMEM) containing 12% fetal calf serum into Hanks' balance salt solution. In DMEM, the 60 kDa protein showed diffused immunofluorescence pattern, and immunoelectron microscopy suggested that this protein was located on the extracellular side of the plasma membrane. After inducing autophagy, the immunofluorescence configuration of the 60 kDa protein changed from the diffused pattern to a granulous one. Immunoelectron microscopy showed that the 60 kDa protein was localized on the luminal side of the limiting membrane of autolysosomes and endosomes. In the presence of bafilomycin A1 which prevents fusion between autophagosomes and lysosomes, the 60 kDa protein was localized on the limiting membrane of the autophagosomes and endosomes. These results suggest that the 60 kDa protein is transported from the plasma membrane to the autophagosome membrane through the endosomes. PMID- 10362070 TI - Co-localization of urokinase and its receptor on established human umbilical vein endothelial cell. AB - Vascular endothelial cells possess antithrombotic properties, which are determined by the balance between plasminogen activators (PAs) and PA inhibitors (PAls). A cell line, TKM-33, has been established and cloned from human umbilical vein endothelial cells, was previously reported to produce a large amount of urokinase-type PA (u-PA) and small amounts of tissue-type plasminogen activator (t-PA) and PA inhibitor-1 (PAI-1). Moreover, TKM-33 expressed the u-PA receptor (u-PAR) which plays an important role in the localization of fibrinolytic activity on cell surface. In the present study, we investigated the localization of u-PA, t-PA, PAI-1 and u-PAR in TKM-33 by using immunofluorescence staining technique. The endothelial cells were strongly stained with anti-PAI-1, anti-u-PA and anti-u-PAR IgGs, and slightly with anti-t-PA IgG. The double immunofluorescence staining with mouse anti-u-PA IgG and rabbit anti-u-PAR IgG followed by rhodamine-conjugated anti-mouse IgG and FITC-conjugated anti-rabbit IgG showed the co-localization of u-PA and u-PAR on the same section of endothelial cells. Although u-PA antigen also existed in the cytoplasm of endothelial cells, u-PAR antigen did not. The treatment of endothelial cells with phorbol-myristate-acetate (PMA) upregulated the expression of u-PA and u-PAR antigens. In this stimulation, u-PAR antigen was detected not only on the surface of the cells but also in the cytoplasm. Thus, the binding of u-PA to u-PAR was confirmed by double immunofluorescence staining. PMID- 10362071 TI - Carnosine stimulates vimentin expression in cultured rat fibroblasts. AB - Two-dimensional electrophoretic gel profiles were compared between rat 3Y1 fibroblasts cultured in the presence and absence of 30 mM L-carnosine (beta alanyl-L-histidine) for one week without any replenishment of medium. While a number of cellular proteins changed their expression levels by the addition of carnosine, we identified one of the most prominently varied proteins as vimentin. Immunoblot analysis with anti-vimentin antibody demonstrated that the vimentin levels increased about 2-fold after one-week culture in the presence of carnosine. We also confirmed that the increase of vimentin expression was dependent on the concentration of carnosine added to the medium. Moreover, when cultured cells were stained with anti-vimentin antibody and observed by light microscopy, most cells grown in the presence of carnosine were found to have markedly developed vimentin filaments. The increase of vimentin expression was also observed by adding with carnosine related dipeptides, N-acetylcarnosine and anserine. PMID- 10362072 TI - Proliferation and differentiation of unilocular fat cells in the bone marrow. AB - Fat cells contribute not only to systemic lipid metabolism, but also to osteogenesis and hemopoiesis in the bone marrow. The present study represents the first attempt to culture mature unilocular fat cells of the bone marrow. Two methods devised in our laboratory were employed: one is the "ceiling culture method" that utilizes the floating property of the cells; the other, a three dimensional collagen gel matrix culture that captures unilocular fat cells in the gel matrix. Using these methods, the proliferation of unilocular fat cells from the bovine metacarpal bone marrow was demonstrated. First, we confirmed the proliferative ability of unilocular fat cells derived from the bone marrow, using autoradiography to study 3H-thymidine incorporation into the nuclei. The unilocular fat cells de-differentiated into fibroblast-like fat cells and then proceeded to proliferate. When they underwent a contact inhibition of growth, re differentiation from fibroblast-like fat cells into unilocular fat cells occurred at a high rate. A specific enzymatic marker of the fat cell, alpha glycerophosphate dehydrogenase activity related to lipogenesis, was then demonstrated in the cultured fat cells. We examined the functional reactivity of the fat cells by treatment with insulin and cyclic-AMP, and both lipogenesis and lipolysis were also confirmed in them. We concluded that unilocular fat cells from the bone marrow de-differentiated, proliferated and re-differentiated in culture. The present results may help to clarify the various functions of fat cells in the bone marrow. PMID- 10362073 TI - The effects of various GTP analogues on microtubule assembly. AB - We synthesized 27 GTP analogues with modification or substitution at positions C2, C6, C8 and ribose moiety to investigate their effect on microtubule (Mt) assembly. It was found that C2 and C6 are both functional for the analogues supporting Mt assembly. It was surprising to find that 2-amino- ATP (n2ATP) substantially supports assembly, and that the appearance of the assembled Mts was indistinguishable from those assembled in the standard GTP assembly buffer solution. Furthermore, 2-amino dATP and dGTP are even more potent than GTP in supporting assembly. The substitution of oxo group at C6 with reactive thiol largely reduced the activity of the analogue to support assembly. When free rotation of the glycosidic linkage of GTP was blocked by the introduction of sulfur atom between C8 and C2' of ribose moiety, it resulted in total suppression of assembly. Purine nucleoside triphosphate was found to support assembly better than GTP, and even more efficient was 2-amino purine nucleoside triphosphate. Interestingly, their deoxy-type analogues were totally inhibitory. Although 2 amino 8-hydroxy ATP and other analogues supported assembly much better than did GTP, their diphosphate analogues were totally incapable of supporting assembly. Finally, bulky fluorescent probes were introduced at C3' of ribose moiety (Mant-8 Br-GTP or Mant-GTP) to visualize the fluorescent signal in assembled Mts. Even in this case, the number of most protofilaments was found to be 14, consistent with that found in Mts assembled in GTP standard buffer solution. PMID- 10362074 TI - Pharmacokinetics of endobronchial tolazoline administration in dogs. AB - Tolazoline is a potent vasodilator of arteries and veins and has a powerful effect on the pulmonary vasculature, reducing hypoxic pulmonary vasoconstriction and lowering pulmonary artery pressure. Intravenous tolazoline lowers the mean pulmonary arterial pressure and resistance and increases the cardiac index when given to infants with persistent pulmonary hypertension (PPHN). Endotracheally administered tolazoline decreases mean pulmonary arterial pressure and pulmonary vascular resistance, and improves oxygenation without the harmful decline in the systemic arterial pressure. The purpose of our study was to examine the pharmacokinetic and pharmacodynamic characteristics of endobronchial tolazoline to determine the relationship between endobronchial tolazoline administration, plasma concentration, and its effects on the cardiovascular and respiratory systems. Tolazoline was administered endobronchially to seven dogs, and its serum concentration and the hemodynamic parameters were monitored for 270 min postdelivery. It was found that 15 sec after dosing, tolazoline plasma concentrations started to increase significantly above baseline levels, reaching a maximum of 9.3+/-8.0 microg x mL(-1) The volume of distribution was 1657+/-321 mL x kg(-1) after 1 2.4+/-1 6.6 min. The extent of tolazoline absorption was 319+/-38 microg x min(-1) mL(-1). The total body clearance was 10.9 +/-4.8 mL x min(-1) x Kg(-1) and the elimination half-life was 156+/-81 min. Endobronchial tolazoline produced an initial short-lived decrease in the mean blood pressure in all the dogs, but thereafter the blood pressure increased gradually above baseline levels. Immediately following endobronchial tolazoline a significant tachycardia developed, peaking at 90 min. Subsequently, the heart rate gradually decreased and stabilized at values above baseline for 200 min. We conclude that an endobronchial bolus dose of tolazoline is effectively absorbed, produces measurable pharmacological effects, and may be beneficial in the therapy of persistent pulmonary hypertension of the newborn. PMID- 10362075 TI - Variations in regional cerebral blood volume in neonates associated with nursery care events. AB - This pilot study investigated the frequency of events that cause cerebral oxygenation disturbances in ventilator-dependant neonates in the neonatal intensive care unit. Continuous, noninvasive, near infrared spectrophotometry measurements (changes in oxy and deoxy hemoglobin concentration, and cytochrome C oxidase redox status) were made at half-second intervals on 10 ventilator dependent neonates (30.5 weeks average corrected gestation age, 1051 g average weight) and annotated to nursery events at the bedside. Examples of disturbances affecting cerebral oxygenation were opening incubator doors, handling, heel stabs, conversation, blanket tucking, and steno-paging. These events produced 7 40% changes in blood volume for durations of 5-60 sec, and occurred at a rate of up to 45 events within a 2 hr period. Spectrometry detected 63% more events than were observed and documented clinically. Noninvasive monitoring of cerebral oxygenation status could give new insight into managing the high-risk infant environment. PMID- 10362076 TI - The heat moisture exchange device (HME) in neonatal ventilation. AB - The objective of this study was to compare the safety and efficacy of the Heat Moisture Exchange (HME) device with conventional humidification in neonates. Sixty-four neonates were randomized at intubation to receive conventional humidification (CH) (n = 34) or HME, via a Neoaid device (n = 30). Groups were compared for intrinsic characteristics and outcome variables. Data were assumed nonparametric and analyzed by Mann-Whitney and Fisher's Exact test. No significant differences were found in group characteristics or outcome variables. Trends were noted for documented patent ductus arteriosus (PDA), endotracheal tube blockage, and positive endotracheal aspirate culture. Rate of pneumothorax 2/34 (CH) versus 4/30 (HME) (p = 0.4); rate of tube blockage 3/34 (CH) versus 2/30 (HME) (p = 1.0); rate of PDA 8/34 (CH) versus 14/30 (HME) (p = 0.093) and rate of endotracheal colonisation 1 7/34 (CH) versus 9/30 (HME) (p = 0.17). No significant difference was found for duration of ventilation or period in greater than 40% oxygen between the two groups. There were no significant outcome differences between CH and HME. The HME device was cost-effective and simple to use. A larger multicenter trial is warranted to confirm the efficacy of HME. PMID- 10362077 TI - Transient hypertrophic cardiomyopathy in the newborn following multiple doses of antenatal corticosteroids. AB - Postnatal exposure to steroids has been associated with hypertrophic cardiomyopathy (HCM) in the newborn. Such an effect has not been described in infants born to mothers who received antenatal steroids. We report three newborns whose mothers were treated with betamethasone prenatally in different doses, duration of time, and who developed various degrees of HCM diagnosed by echocardiography. There was no maternal evidence of diabetes except for one infant whose mother had a normal fasting and post-prandial blood glucose prior to steroid therapy, but an abnormal one hour postprandial glucose after 8 weeks of betamethasone therapy, with a normal HbA1 C level. There was no family history of HCM, no history of maternal intake of other relevant medications, and no hypertension in all three newborns. Follow-up echocardiography revealed complete resolution of the HCM changes in all infants. We suggest that repeated antenatal maternal steroid intake may cause changes of HCM in the newborn. These changes appear to be dose- and duration-related and are mostly reversible. Further prospective controlled studies to evaluate these observations and to investigate potential mechanisms are warranted. PMID- 10362078 TI - Forceps and vacuum delivery: expectations of residency and fellowship training program directors. AB - The objective of this study is to compare current forceps training practices in North American obstetrical residency training programs with that in maternal fetal medicine fellowship programs. We sent a survey to all obstetrics and gynecology residency training programs and to all maternal-fetal medicine fellowship programs in North America. After sending out 354 questionnaires, 219 were returned for a response rate of 62%. The response rate for fellowship programs (52 of 59; 88%) was significantly greater than that of residency training programs (167 of 295; 56.6%) (p < 0.05). All fellowship training programs were using the 1988 ACOG forceps classification system, as were 98% of the residency training programs. Eighty-five percent of fellowship directors and 80% of residency directors felt the same system should be used for vacuum deliveries. All residency and fellowship directors expected proficiency with both instruments for outlet deliveries. For low deliveries requiring < or =45 degrees of rotation, at least 92% expected proficiency with both instruments. For low forceps deliveries with >45 degrees of rotation, 82% of fellowship directors and 80% of residency directors expected proficiency. For low-vacuum deliveries with >45 degrees of rotation, 80% of fellowship directors and 76% of residency directors expected proficiency. Significantly more fellowship directors expected midforceps proficiency (47%) than did residency program directors (38%) (p < 0.05). Midvacuum proficiency was expected by 73% of fellowship directors and 69% of residency directors. The ACOG 1988 forceps classification system has now achieved wide acceptance and is taught by both residency and fellowship program directors. Most program directors favor using the same classification system for vacuum extraction deliveries. In general, the expectations of the residency program directors mirror those of maternal-fetal medicine fellowship directors. While outlet and low operations with < or =45 degrees of rotation are taught and proficiency is expected, most programs no longer expect proficiency in midforceps delivery, but do expect proficiency in midvacuum delivery. Proficiency in low operations with rotations < or =45 degrees is still expected. PMID- 10362079 TI - Prader-Willi syndrome associated with fetal goiter: a case report. AB - We describe a unique case of a newborn with Prader-Willi syndrome who presented with fetal goiter as well as neonatal thyroid abnormalities, marked hypotonia, and thrombocytopenia. These new clinical observations may correlate with the uniparental monodisomy form of inheritance of this genetic condition. PMID- 10362080 TI - Obstetrical determinants of neonatal neurological morbidity in < or = 1000-gram infants. AB - The purpose of this study is to identify obstetrical factors associated with adverse neurological outcome in < or =1000-g infants. In a 1-year (1992-1993) observational study, the NICHD MFMU Network collected obstetrical risk factors for 486 infants who weighed < or =1000 g at birth and who survived > 2 days. Infants' records were abstracted for seizures, intraventricular hemorrhage, and an abnormal neurological evaluation. Seventy-nine (16%) infants had a Grade III or IV intraventricular hemorrhage, 46 (9%) developed seizures and 57 (14%) had an abnormal neurological evaluation. Both lower birth weight and earlier gestational age correlated (P <0.01) with an increasing incidence of all three outcomes. Several other factors appeared to be associated with neurological morbidity, however, after controlling for potential confounders in the multivariate analyses, most of these factors were no longer significant. African-American race, odds ratio (OR) 0.6 (0.3-1.0), and severe preeclampsia, OR 0.2 (0.1-0.7), were protective against intraventricular hemorrhage. Maternal treatment with corticosteroids did not impact neurological outcome in this study population. We conclude that, in a population of < or =1000-g infants, lower birth weight and earlier gestational age were the only consistently significant predictors of all three adverse neurological outcomes. PMID- 10362081 TI - Sequential peripartum herpes simplex virus type 2 disease in parents and their newborn mimicking intrafamily spread of common viruses. AB - Herpes simplex type 2 (HSV2) disease developed sequentially among two parents and their newborn. The father first became ill with upper-respiratory symptoms and fever. Then, 5 days later, shortly after delivery, the mother had fever, pharyngitis, and diarrhea. Subsequently, the infant developed undifferentiated febrile illness at the age of 3 days. HSV etiology was recognized by incidental isolation of HSV2 from the newborn naospharynx. The father never developed genital lesions and the mother's symptoms remained nonspecific for several days prior to the onset of genital manifestations. The sequential emergence and manifestations of these infections could have been misconstrued for an intrafamily spread of respiratory or enteric viruses. This cluster illustrates that HSV2 may cause sequential symptomatic disease in susceptible individuals mimicking other viruses. PMID- 10362082 TI - Prenatal diagnosis and perinatal management of fetal sacrococcygeal teratoma. AB - Sacrococcygeal teratoma is the most common fetal neoplasm, with an incidence of 1 in 40,000 births. Fetuses with this malformation are at risk for significant perinatal morbidity and mortality. We identified nine fetuses with sacrococcygeal teratomas that were diagnosed antenatally and managed at the University of North Carolina Hospitals over a 7-year period. We retrospectively reviewed the charts of mothers and infants and recorded data concerning perinatal and surgical management. Six infants survived the neonatal period. All infants diagnosed after 20 weeks' gestation survived. Fetal hydrops developed in three fetuses, all of whom died. Inadequate ventilation secondary to prematurity was a contributing factor in each lethal case. Diagnosis at an early gestational age, development of fetal hydrops, and premature delivery predicted a poor prognosis. When possible, we recommend that delivery be delayed to allow for fetal development. Stabilization of the infant should be attempted before resection of the teratoma. PMID- 10362083 TI - Surgery and cancer: opinion, evidence, and proof. PMID- 10362084 TI - Prognostic factors in superficial adult soft tissue sarcomas: analysis of a series of 105 patients. AB - BACKGROUND AND OBJECTIVES: This study was undertaken to study the behavior of superficial soft tissue sarcomas (STS) and determine the factors related to prognosis. METHODS: The clinical records of 105 adults (56 men, 49 women, mean age: 56.4 years) were retrospectively analyzed. Univariate analysis was performed on the entire group for overall survival (OS), and metastasis-free survival (MFS). Local recurrence-free survival (LRFS) was studied only on patients first treated in our institute. RESULTS: With a median follow-up of 111.9 months, 66 (62.9%) patients were alive; 25 (23.8%) had died of their disease. For the entire series, 10-year OS and MFS were 62.5% and 71.9% respectively. For fifty-two patients treated for their sarcoma at the Institution since the first tumor occurrence event, 10-year LRFS was 80%. Tumor grade was the only factor correlated with OS and MFS, while tumor size was the main factor correlated with LRFS. CONCLUSION: Tumor size affects local control in STS while tumor grade is correlated with OS and MFS. PMID- 10362085 TI - Cellular manifestations of human papillomavirus infection in the oral mucosa. AB - BACKGROUND AND OBJECTIVES: Human papillomavirus infection has been suggested to play a role in the development of epithelial carcinomas, particularly those of the uterine cervix. Less information is available on the role of the virus in oral lesions. It has been proposed that the viral oncoproteins specifically complex with the products of cellular tumor suppressor gene, namely E6 with p53 and E7 with retinoblastoma gene product. Inactivation or mutation in p53 gene is also known to result in loss of control over the cell cycle and increases in tumor proliferation rates. The present study examines the role of HPV infection in relation to p53 and the extent of the tumor proliferative compartment reflected by cyclin D1 and Ki-67 expression during various phases of tumor progression in the oral epithelium. METHOD: Nonisotopic in situ hybridization (NISH) was performed to detect HPV 6/11 and 16/18. Expression of p53, cyclin D1, Ki-67, and the HPV 16/18 E6 protein were detected by immunocytochemistry. RESULTS: There was significant correlation between the extent of histological abnormality and HPV infection. A correlation (r = 0.250, P = 0.0089) was evident between the presence of HPV 16 and occurrence of invasive cancer. Expression of the tumor suppressor p53 protein also showed significant positive correlation with histology (r = 0.475, P = 0.00004). The tumor proliferative fraction also increased with the extent of histological abnormality (r = 0.387, P = 0.0003 for cyclin D1 and r = 0.463, P = 0.0001 for Ki 67). Accumulation of p53 and increase in tumor proliferation also correlated to the presence of HPV infection (r = 0.511, P = 0.00003 for p53; r = 0.478, P = 0.00002 for cyclin D1 and r = 0.521, P = 0.00004 for Ki-67). CONCLUSIONS: The present study thus demonstrates the importance of HPV infection in oral tissue. Expression of the high-risk HPV 16/18 E6 protein also appears to be a critical event along with aberrant p53 expression. These results are of significance to the molecular epidemiology of oral cancer and may also be used to supplement and elaborate the diagnosis of oral lesions. PMID- 10362086 TI - Induction of human leukocyte antigen (HLA)-A2-restricted and MAGE-3-gene-derived peptide-specific cytolytic T lymphocytes using cultured dendritic cells from an HLA-A2 esophageal cancer patient. AB - BACKGROUND AND OBJECTIVES: Using peripheral blood mononuclear cells (PBMCs) from a 10-year survivor with established human leukocyte antigen (HLA)-A2(+) and MAGE 3(+) esophageal cancer cell line (KYSE-170), we examined the induction of HLA-A2 restricted and MAGE-3-gene-derived peptide (FLWGPRALV, amino acids 271-279) specific cytolytic T lymphocytes (CTLs). METHODS: Autologous dendritic cells (DCs) cultured with granulocyte-macrophage colony stimulating factor and interleukin-4 were used as antigen presenting cells. PBMCs were stimulated by peptide-pulsed DCs in vitro. RESULTS: PBMC cocultured with FLWGPRALV-pulsed DCs could induce the relevant peptide-specific CTLs, which had tumor necrosis factor production and specific cytotoxicity against relevant peptide-pulsed autologous DCs (34%, effector:target ratio = 40:1). Moreover, they showed specific cytotoxicity against the autologous esophageal cancer cell line KYSE-170 (17%, effector:target ratio = 40:1). CONCLUSIONS: These results suggest that FLWGPRALV pulsed cultured DCs would be a potent candidate for peptide vaccine against HLA A2(+) and MAGE-3(+) esophageal cancer. PMID- 10362088 TI - Bronchogenic carcinoma in young patients. AB - BACKGROUND AND OBJECTIVES: Some investigators have suggested that lung cancer in young patients has a more aggressive course and poorer prognosis than lung cancer in older patients. METHODS: A retrospective review is presented of patients less than 40 years of age with bronchogenic carcinoma treated at Roswell Park Cancer Institute between 1984 and 1994, with comparison to a cohort of patients treated in the previous decade. RESULTS: There were 76 patients (41 male and 35 female). Mean age was 35 years (range, 26-39). Adenocarcinoma in 33 patients (43%) and undifferentiated large-cell carcinoma in 22 patients (29%) were the predominant histologic types. Stage IIIa or greater disease was present in 63 (83%) patients. Treatment consisted of chemotherapy (55 patients), radiation therapy (54 patients), and surgery (33 patients). Surgical procedures included pneumonectomy (14 patients), lobectomy (11 patients), wedge resection (1 patient), and thoracotomy only for unresectable disease (7 patients). Operative mortality was 6% (two patients who had radical pneumonectomy for T4 cancer). Median survival for the entire group of patients was 10.4 months, and 5-year survival was 8%. Univariate analysis identified acute presentation (P = 0.02), no resection (P = 0.0001), and higher stage (P = 0.0001) as negative prognostic factors. On multivariate analysis, stage of disease was the only independent predictor of survival (P = 0.005). Resectability was slightly higher (34%, 26/76, vs. 21%, 19/89; P = 0.06) and survival was marginally better (median 10.4 vs. 7.5 months; P = 0.05) than that seen at our institution in the previous decade. CONCLUSIONS: Young patients with lung cancer often have advanced disease at the time of presentation. Nevertheless, they should be treated in accordance with standard stage-specific treatment guidelines. PMID- 10362087 TI - Prognostic significance of CD44 and nm23 expression in patients with stage II and stage IIIA gastric carcinoma. AB - BACKGROUND AND OBJECTIVES: Predicting the prognosis in gastric carcinoma patients with intermediate stages is difficult. We investigated the prognostic impacts of CD44 and nm23 expression in a homogeneous group of patients with stage II and IIIA gastric carcinoma who had undergone curative resections. METHODS: A total of 261 paraffin-embedded gastric carcinomas were stained with the monoclonal antibodies CD44 and nm23 using the labeled streptovidin biotin method. RESULTS: The expression of CD44 and nm23 was detected, respectively, in 31.0% (81/261) and 70.1% (183/261) of all tumors. There was no correlation between CD44 expression and clinicopathological variables. However, nm23 was more frequently expressed in older patients with differentiated adenocarcinoma. A significant difference in 5 year survival rates was found between patients with CD44-positive (43.2%) and CD44-negative tumors (63.4%), (P = 0.0018). However, there was no significant difference in 5-year survival rates between patients with nm23-positive (54.7%) and nm23-negative tumors (62.7%) (P = 0.2734). CONCLUSIONS: CD44 expression was a significant adverse prognostic factor in gastric carcinoma and may be a predictor of metastatic potential of the primary tumor. By contrast, immunohistochemical detection of nm23 expression was not a predictor of outcome of patients with gastric carcinoma. PMID- 10362089 TI - Prognostic factors in retroperitoneal sarcomas: ploidy of DNA as a predictor of clinical outcome. AB - BACKGROUND AND OBJECTIVES: Radical surgery is the best mode of treatment of retroperitoneal sarcomas (RS); however, common recurrences are unpredictable. METHODS: For the better understanding of outcomes and possibilities of treatment retrospective analysis of different factors, including DNA content, was performed based on 70 patients. RESULTS: Leiomyosarcoma and liposarcoma were most common histologic types of classified sarcomas. Different kinds of resection were successfully performed in 51 patients (73%) and 35 of their available DNA specimens were analyzed. The actuarial 5- and 10-year survival rates in the resection group were 53% and 40%, respectively, with the median survival of 57 months. Patients with diploid resected tumors had a better 10-year survival rate (58%), than those patients with aneuploid tumors (25%,)--P<0.005. Those patients with low-grade sarcomas had a significantly longer survival than those with high grade sarcomas (10-year survival rate: 44% compared to 29%). In the univariate analysis, adjuvant therapy, type of histology, type of surgery, location of tumor, and S-phase fraction had no influence on survival. In the multivariate analysis (Cox), only ploidy was an independent prognostic variable. Relative risk of death was over three times higher for aneuploid than for diploid tumors. CONCLUSION: Tumor ploidy should be analyzed in every case of retroperitoneal sarcoma for better assessment of prognosis and possible indication for adjuvant therapy. PMID- 10362090 TI - MCA106 fibrosarcoma cells transduced with granulocyte/macrophage colony stimulating factor are not superior to the wild-type cells in suppressing the growth of hepatic metastases. AB - BACKGROUND AND OBJECTIVES: Vaccination with cytokine gene-modified tumor cells augments the immune response against established tumors and protects against tumor challenges. In this study, we investigated the vaccine potential of GM-CSF transduced MCA106 fibrosarcoma (MCA-GMCSF) cells in the C57BL/6 (B6) murine hepatic metastasis model. METHODS: Experimental mice received one to three weekly vaccines (subcutaneous/intramuscular, s.c./i.m.) of irradiated, parental, or GM CSF-transduced MCA106 tumor cells. One week after the last immunization, hepatic metastases were established through the intrasplenic injection of live MCA106 parental (MCA106P) tumor cells. The animals were then sacrificed 3-4 weeks after surgery for evaluation of hepatic tumor burden. RESULTS: Based on in vivo experiments, both GM-CSF-modified and parental MCA106 tumor cell vaccines induced strong protection against hepatic tumor growth with grossly visible tumors rarely identified. This protection was evident even at a single vaccine dose of as low as 1x10(5) irradiated cells. Unimmunized control mice, on the other hand, consistently developed substantial hepatic tumors. Cytotoxicity assays on splenocytes (cultured in vitro for 4-5 days) showed that both groups of vaccinated mice developed strong tumor-specific cytotoxic T-lymphocyte (CTL) responses. Immunohistochemical analysis of injection sites showed infiltration of dendritic cells (DCs) and macrophages into subcutaneously injected MCA-GMCSF cells. Mostly macrophages, however, were seen at the injection site of MCA106P cells. Furthermore, the MCA106P cells expressed high levels of MHC class I antigens and the level of expression was not significantly altered by transduction with the GM-CSF gene. The high expression of MHC class I antigens probably contributed to the strong immunogenicity of the MCA106P cell vaccine. CONCLUSIONS: This study demonstrates that MCA106 parental cells are as effective as the GM-CSF-transduced cells in suppressing the growth of hepatic metastases. The cellular immune responses induced by these two vaccines, however, are probably mediated by different subsets of host effector cells. These results have important implications for the use of GM-CSF-transduced cell vaccines in the immunotherapy of tumors that have the propensity to metastasize through the lymphatic channels and the circulatory system. PMID- 10362091 TI - Accuracy of biopsy and cytology for the preoperative diagnosis of colorectal adenocarcinoma. AB - BACKGROUND AND OBJECTIVES: Endoscopic biopsy for the diagnosis of colorectal adenocarcinoma is not accurate in every case. Brush cytology can increase the sensitivity for the diagnosis of gastroesophageal lesions when combined with biopsy, but very little information is available for these techniques in the diagnosis of colorectal adenocarcinoma. METHODS: A retrospective medical records review of 110 patients was performed. All patients underwent a colorectal resection for primary adenocarcinoma after a diagnostic endoscopy. Biopsy and brush cytology was evaluated for their respective sensitivity. Seventy-three patients had both biopsy and cytology. RESULTS: The sensitivity of biopsy was 83.6% (92/110); for cytology, 78.1% (57/73; P = 0.44). From the 73 patients who had both diagnostic techniques, 68.5% (50/73) had both positive biopsy and cytology, 12.3% (9/73) only a positive biopsy, and 9.6% (7/73) only a positive cytology. The two techniques combined were not significantly superior to biopsy alone (90.4%, 66/73, vs. 80.8%, 59/73, respectively; P = 0.16), but tended to be superior to cytology alone (P = 0.07). CONCLUSIONS: Cytology and biopsy have a comparable sensitivity. The combination of the two techniques compares favorably, but does not significantly increase the sensitivity of biopsy alone. Both techniques should be used whenever there are any uncertainties concerning the diagnosis of colorectal adenocarcinoma. PMID- 10362092 TI - Incidental liposarcomas identified during hernia repair operations. AB - BACKGROUND AND OBJECTIVES: Since the inguinal region communicates with the retroperitoneum, both retroperitoneal as well as de novo spermatic cord liposarcomas may be detected during hernia repair operations. We assessed the incidence of liposarcomas presenting at hernia repair in our hospital. METHODS: We performed a clerical review of pathology reports on adult tissue accessioned during hernia repair operations and reviewed operating room logs to obtain information concerning the total number of hernia repair operations (since some operations afford no accessioned tissue). RESULTS: Between 1992 and 1997, 1,736 adult hernia repair specimens were accessioned from approximately 2,000 operations. Among these, 22% had an associated cord lipoma; 2 cases were well differentiated liposarcomas. These were from males aged 56 and 64 years in contrast to the mean age of 35 years for cord lipoma and measured 13 and 10 cm, whereas the mean size for cord lipomas was 5.5 cm. One of the liposarcomas had radiographic evidence of extension from a retroperitoneal lesion; the other appeared confined to the groin. On surgical exploration, the lesion was restricted to the spermatic cord region in both cases despite the suggestion of retroperitoneal extension/involvement in one. CONCLUSIONS: Incidental liposarcomas identified during hernia operations are rare (<0.1% at our institution) but their presence merits histologic evaluation of adipose tissue from these cases. However, if efforts to contain costs are implemented and histologic review of such tissue is deemed generally unrewarding, large (>10 cm) fatty masses from this area should still be sampled. PMID- 10362093 TI - Esophageal mucoepidermoid carcinoma containing signet-ring cells: three case reports and a literature review. AB - We report 3 cases of esophageal signet-ring cell carcinoma which were found in a set of 505 resected esophageal tumors. The incidence of esophageal signet-ring cell carcinoma was 0.6%. All of the signet-ring cell carcinomas were histologically mixed with squamous cell carcinoma (mucoepidermoid carcinoma). The signet-ring cells had intracellular mucin, which was positive for both periodic acid-Schiff (PAS) and alcian blue at pH 2.5. At the time of presentation, extensive extraesophageal tumor spread and local extension were found in all cases. All of the patients died within 2 years after the esophagectomy irrespective of whether they received chemotherapy or radiotherapy. Our results, and those previously reported, suggest that most esophageal carcinomas containing signet-ring cell carcinoma are aggressive neoplasms associated with a poor prognosis after esophagectomy. PMID- 10362094 TI - Tumor-induced osteomalacia and symptomatic looser zones secondary to mesenchymal chondrosarcoma. AB - Tumor-induced osteomalacia is a rare clinical entity that is associated with soft tissue or skeletal tumors. We present a case report of a patient with a chest wall mesenchymal chondrosarcoma who presented with bone pain. The patient had skeletal changes in the femoral neck and fibula consistent with osteomalacia and laboratory values suggesting phosphate diabetes. The patient was treated with tumor resection and phosphate supplementation with reversal of the signs and symptoms of osteomalacia. Tumor-induced osteomalacia is vitamin-D-resistant and often reversed by complete removal of the tumor. Most commonly, the causative tumors are of vascular, mesenchymal, or fibrous origin. The osteomalacia is associated with bone pain, muscle weakness, and radiographic changes. Tumor induced humoral factors have been implicated in causing the osteomalacia, but the definite etiology has yet to be determined. Current treatment includes complete tumor resection and electrolyte supplementation. PMID- 10362095 TI - Reconstruction after larynx-preserving extensive tracheal resection with axillofemoral bypass grafting and free skin flap implantation. PMID- 10362096 TI - Modified hemipelvectomy. PMID- 10362097 TI - Helicobacter pylori and gastric cancer: time for mega-trials? PMID- 10362098 TI - Endogenous sex hormones and prostate cancer: a quantitative review of prospective studies. AB - This paper presents a quantitative review of the data from eight prospective epidemiological studies, comparing mean serum concentrations of sex hormones in men who subsequently developed prostate cancer with those in men who remained cancer free. The hormones reviewed have been postulated to be involved in the aetiology of prostate cancer: androgens and their metabolites testosterone (T), non-SHBG-bound testosterone (non-SHBG-bound T), di-hydrotestosterone (DHT), androstanediol glucuronide (A-diol-g), androstenedione (A-dione), dehydroepiandrosterone sulphate (DHEAS), sex hormone binding globulin (SHBG), the oestrogens, oestrone and oestradiol, luteinizing hormone (LH) and prolactin. The ratio of the mean hormone concentration in prostate cancer cases to that of controls (and its 95% confidence interval (CI)) was calculated for each study, and the results summarized by calculating the weighted average of the log ratios. No differences in the average concentrations of the hormones were found between prostate cancer cases and controls, with the possible exception of A-diol-g which exhibited a 5% higher mean serum concentration among cases relative to controls (ratio 1.05, 95% CI 1.00-1.11), based on 644 cases and 1048 controls. These data suggest that there are no large differences in circulating hormones between men who subsequently go on to develop prostate cancer and those who remain free of the disease. Further research is needed to substantiate the small difference found in A-diol-g concentrations between prostate cancer cases and controls. PMID- 10362099 TI - Induction of matrix metalloprotease-1 gene expression by retinoic acid in the human pancreatic tumour cell line Dan-G. AB - We have investigated the effects of retinoic acid (RA) on matrix metalloprotease 1 (MMP-1) gene expression in the human pancreatic tumour cell line Dan-G. 13-cis RA results in a time- and dose-dependent increase of MMP-1 protein concentration. These stimulatory effects were paralleled by a time- and dose-dependent increase of MMP-1 mRNA steady-state concentrations. Nuclear run-on analysis revealed that the increase of MMP-1 mRNA was partially due to an increase of MMP-1 gene transcription. In addition, 13-cis RA treatment results in an increase of MMP-1 mRNA stability. These data demonstrate that RA stimulates MMP-1 gene expression in human pancreatic carcinoma cells by transcriptional and post-transcriptional mechanisms. PMID- 10362100 TI - Expression levels of the DNA repair enzyme HAP1 do not correlate with the radiosensitivities of human or HAP1-transfected rat cell lines. AB - Apurinic/apyrimidinic (AP) sites in DNA are potentially lethal and mutagenic. They can arise spontaneously or following DNA damage from reactive oxygen species or alkylating agents, and they constitute a significant product of DNA damage following cellular exposure to ionizing radiation. The major AP endonuclease responsible for initiating the repair of these and other DNA lesions in human cells is HAP1, which also possesses a redox function. We have determined the cellular levels of this enzyme in 11 human tumour and fibroblast cell lines in relation to clonogenic survival following ionizing radiation. Cellular HAP1 levels and surviving fraction at 2 Gy (SF2) varied five- and tenfold respectively. However, no correlation was found between these two parameters following exposure to gamma-irradiation at low (1.1 cGy per min) or high (108 cGy per min) dose rates. To examine this further, wild-type and mutant versions of HAP1 were overexpressed, using an inducible HAP1 cDNA expression vector system, in the rat C6 glioma cell line which has low endogenous AP endonuclease activity. Induction of wild-type HAP1 expression caused a > fivefold increase in the capacity of cellular extracts to cleave an oligonucleotide substrate containing a single abasic site, but increased expression did not confer increased resistance to gamma-irradiation at high- or low-dose rates, or to the methylating agent methyl methanesulphonate (MMS). Expression in C6 cell lines of mutant forms of HAP1 deleted for either the redox activator or DNA repair functions displayed no apparent titrational or dominant negative effects. These studies suggest that the levels of endogenous AP endonuclease activities in the various cell lines examined are not limiting for efficient repair in cells following exposure to ionizing radiation or MMS. This contrasts with the correlation we have found between HAP1 levels and radiosensitivity in cervix carcinomas (Herring et al (1998) Br J Cancer 78: 1128-1133), indicating that HAP1 levels in this case assume a critical survival role and hence that established cell lines might not be a suitable model for such studies. PMID- 10362101 TI - BPD-MA-mediated photosensitization in vitro and in vivo: cellular adhesion and beta1 integrin expression in ovarian cancer cells. AB - Benzoporphyrin derivative monoacid (BPD-MA) photosensitization was examined for its effects on cellular adhesion of a human ovarian cancer cell line, OVCAR 3, to extracellular matrix (ECM) components. Mild BPD-MA photosensitization (approximately 85% cell survival) of OVCAR 3 transiently decreased adhesion to collagen IV, fibronectin, laminin and vitronectin to a greater extent than could be attributed to cell death. The loss in adhesiveness was accompanied by a loss of beta1 integrin-containing focal adhesion plaques (FAPs), although beta1 subunits were still recognized by monoclonal antibody directed against human beta1 subunits. In vivo BPD-MA photosensitization decreased OVCAR 3 adhesiveness as well. Photosensitized adhesion was reduced in the presence of sodium azide and enhanced in deuterium oxide, suggesting mediation by singlet oxygen. Co localization studies of BPD-MA and Rhodamine 123 showed that the photosensitizer was largely mitochondrial, but also exhibited extramitochondrial, intracellullar, diffuse cytosolic fluorescence. Taken together, these data show that intracellular damage mediated by BPD-PDT remote from the FAP site can affect cellular-ECM interactions and result in loss of FAP formation. This may have an impact on long-term effects of photodynamic therapy. The topic merits further investigation. PMID- 10362102 TI - Synergistic induction of ICAM-1 expression by cisplatin and 5-fluorouracil in a cancer cell line via a NF-kappaB independent pathway. AB - Cisplatin (CDDP) and 5-fluorouracil (5-FU) are common anti-tumour agents, and the anti-tumour effect of CDDP and 5-FU are synergistically enhanced by combined treatment. To clarify the mechanisms of this synergism, we examined the effect of CDDP and 5-FU on the expression of cell adhesion molecules involved in recognition of cancer cells by T lymphocytes. When NA cells, a squamous cell carcinoma cell line, were exposed to CDDP and 5-FU for 18 h, the expression of intercellular adhesion molecule-1 (ICAM-1) was synergistically induced, whereas CDDP or 5-FU alone did not induce the expression of ICAM-1, as determined by flow cytometry. Expression of ICAM-2 and ICAM-3, which are recognized by the same counter receptor on T-cells, were not up-regulated by CDDP and 5-FU. RT-PCR analysis showed that the induction of ICAM-1 on NA cells might be due to transcriptional induction of ICAM-1 mRNA. Treatment with genistein, a protein tyrosine kinase (PTK) inhibitor, inhibited the induction of ICAM-1 on NA cells by CDDP and 5-FU, whereas staurosporin, a protein kinase C inhibitor, did not. Although CDDP and 5-FU induced binding at the nuclear factor kappa B (NF-kappaB) site in the ICAM-1 promoter, pretreatment with genistein did not prevent CDDP and 5-FU-induced binding at the NF-kappaB site. Moreover, a NF-kappaB nuclear translocation inhibitor did not inhibit the induction of ICAM-1 expression by treatment with CDDP and 5-FU. The synergistic effect of CDDP and 5-FU was not specific to NA cells, since ICAM-1 was synergistically induced by CDDP and 5-FU on HSC-4 cells, a squamous cell carcinoma cell line. These findings indicate that treatment with CDDP and 5-FU induces ICAM-1 expression by a NF-kappaB independent regulatory mechanism involving PTK. PMID- 10362103 TI - Enhanced delivery of carboplatin into brain tumours with intravenous Cereport (RMP-7): dramatic differences and insight gained from dosing parameters. AB - Cereport (RMP-7) is a selective bradykinin B2 receptor agonist which increases the permeability of the 'blood-brain tumour barrier' (BBTB) to increase delivery of chemotherapeutic agents to brain tumours. A series of experiments was performed in an RG2 rodent model of glioma to evaluate and refine intravenous (i.v.) parameters to optimize Cereport's clinical utility. The first experiment demonstrated that while carboplatin levels were increased by twofold when given as a bolus during the Cereport infusion, no increase in carboplatin levels were seen when Cereport and carboplatin were simultaneously co-infused for 15 min. A subsequent experiment established that a major factor responsible for the lack of an effect with the co-infusion paradigm was tachyphylaxis to Cereport during the 15 min infusion, for a progressively diminished response to Cereport occurred over that time frame, as plasma levels of carboplatin were rising. A final experiment adjusted the timing of the Cereport and carboplatin infusions so that higher plasma carboplatin levels were achieved prior to initiating the Cereport infusion. Significant uptake effects were achieved when the carboplatin infusion preceded the Cereport infusion by 10 min (i.e. 5 min overlap in the delivery of the two agents). Collectively, these data provide the first systematic evaluation of dosing parameters involving receptor-mediated changes in BBTB permeability and provide new information regarding the pharmacodynamics and potential clinical use of Cereport. PMID- 10362104 TI - Mode of action of thiocoraline, a natural marine compound with anti-tumour activity. AB - Thiocoraline, a new anticancer agent derived from the marine actinomycete Micromonospora marina, was found to induce profound perturbations of the cell cycle. On both LoVo and SW620 human colon cancer cell lines, thiocoraline caused an arrest in G1 phase of the cell cycle and a decrease in the rate of S phase progression towards G2/M phases, as assessed by using bromodeoxyuridine/DNA biparametric flow cytometric analysis. Thiocoraline does not inhibit DNA topoisomerase II enzymes in vitro, nor does it induce DNA breakage in cells exposed to effective drug concentrations. The cell cycle effects observed after exposure to thiocoraline appear related to the inhibition of DNA replication. By using a primer extension assay it was found that thiocoraline inhibited DNA elongation by DNA polymerase alpha at concentrations that inhibited cell cycle progression and clonogenicity. These studies indicate that the new anticancer drug thiocoraline probably acts by inhibiting DNA polymerase alpha activity. PMID- 10362105 TI - Mechanisms of synergism between cisplatin and gemcitabine in ovarian and non small-cell lung cancer cell lines. AB - 2',2'-Difluorodeoxycytidine (gemcitabine, dFdC) and cis-diammine-dichloroplatinum (cisplatin, CDDP) are active agents against ovarian cancer and non-small-cell lung cancer (NSCLC). CDDP acts by formation of platinum (Pt)-DNA adducts; dFdC by dFdCTP incorporation into DNA, subsequently leading to inhibition of exonuclease and DNA repair. Previously, synergism between both compounds was found in several human and murine cancer cell lines when cells were treated with these drugs in a constant ratio. In the present study we used different combinations of both drugs (one drug at its IC25 and the other in a concentration range) in the human ovarian cancer cell line A2780, its CDDP-resistant variant ADDP, its dFdC resistant variant AG6000 and two NSCLC cell lines, H322 (human) and Lewis lung (LL) (murine). Cells were exposed for 4, 24 and 72 h with a total culture time of 96 h, and possible synergism was evaluated by median drug effect analysis by calculating a combination index (CI; CI < 1 indicates synergism). With CDDP at its IC25, the average CIs calculated at the IC50, IC75 IC90 and IC95 after 4, 24 and 72 h of exposure were < 1 for all cell lines, indicating synergism, except for the CI after 4 h exposure in the LL cell line which showed an additive effect. With dFdC at its IC25, the CIs for the combination with CDDP after 24 h were < 1 in all cell lines, except for the CIs after 4 h exposure in the LL and H322 cell lines which showed an additive effect. At 72 h exposure all CIs were < 1. CDDP did not significantly affect dFdCTP accumulation in all cell lines. CDDP increased dFdC incorporation into both DNA and RNA of the A2780 cell lines 33- and 79-fold (P < 0.01) respectively, and tended to increase the dFdC incorporation into RNA in all cell lines. In the AG6000 and LL cell lines, CDDP and dFdC induced > 25% more DNA strand breaks (DSB) than each drug alone; however, in the other cell lines no effect, or even a decrease in DSB, was observed. dFdC increased the cellular Pt accumulation after 24 h incubation only in the ADDP cell line. However, dFdC did enhance the Pt-DNA adduct formation in the A2780, AG6000, ADDP and LL cell lines (1.6-, 1.4-, 2.9- and 1.6-fold respectively). This increase in Pt-DNA adduct formation seems to be related to the incorporation of dFdC into DNA (r = 0.91). No increase in DNA platination was found in the H322 cell line. dFdC only increased Pt-DNA adduct retention in the A2780 and LL cell lines, but decreased the Pt-DNA adduct retention in the AG6000 cell line. In conclusion, the synergism between dFdC and CDDP appears to be mainly due to an increase in Pt-DNA adduct formation possibly related to changes in DNA due to dFdC incorporation into DNA. PMID- 10362106 TI - Alpha-bromoacryloyl derivative of distamycin A (PNU 151807): a new non-covalent minor groove DNA binder with antineoplastic activity. AB - PNU 151807 is a new synthetic alpha-bromoacryloyl derivative of distamycin A. In the present study we investigated the DNA interaction and the mechanism of action of this compound in parallel with the distamycin alkylating derivative, tallimustine. PNU 151807 possesses a good cytotoxic activity in in vitro growing cancer cells, even superior to that found for tallimustine. By footprinting experiments we found that PNU 151807 and tallimustine interact non-covalently with the same AT-rich DNA regions. However, differently from tallimustine, PNU 151807 failed to produce any DNA alkylation as assessed by Taq stop assay and N3 or N7-adenine alkylation assay in different DNA sequences. PNU 151807, like tallimustine, is able to induce an activation of p53, and consequently of p21 and BAX in a human ovarian cancer cell line (A2780) expressing wild-type p53. However, disruption of p53 function by HPV16-E6 does not significantly modify the cytotoxic activity of the compound. Flow cytometric analysis of cells treated with equitoxic concentrations of PNU 151807 and tallimustine showed a similar induction of accumulation of cells in the G2 phase of the cell cycle but with a different time course. When tested against recombinant proteins, only the compound PNU 151807 (and not tallimustine or distamycin A) is able to abolish the in vitro kinase activity of CDK2-cyclin A, CDK2-cyclin E and cdc2-cyclin B complexes. The results obtained showed that PNU 151807 seems to have a mechanism of action completely different from that of its parent compound tallimustine, possibly involving the inhibition of cyclin-dependent kinases activity, and clearly indicate PNU 151807 as a new non-covalent minor groove binder with cytotoxic activity against cancer cells. PMID- 10362107 TI - Delta-aminolaevulinic acid-induced photodynamic therapy inhibits protoporphyrin IX biosynthesis and reduces subsequent treatment efficacy in vitro. AB - Recently, considerable interest has been given to photodynamic therapy of cancer using delta-aminolaevulinic acid to induce protoporphyrin IX as the cell photosensitizer. One advantage of this modality is that protoporphyrin IX is cleared from tissue within 24 h after delta-aminolaevulinic acid administration. This could allow for multiple treatment regimens because of little concern regarding the accumulation of the photosensitizer in normal tissues. However, the haem biosynthetic pathway would have to be fully functional after the first course of therapy to allow for subsequent treatments. Photosensitization of cultured R3230AC rat mammary adenocarcinoma cells with delta-aminolaevulinic acid induced protoporphyrin IX resulted in the inhibition of porphobilinogen deaminase, an enzyme in the haem biosynthetic pathway, and a concomitant decrease in protoporphyrin IX levels. Cultured R3230AC cells exposed to 0.5 mM delta aminolaevulinic acid for 27 h accumulated 6.07 x 10(-16) mol of protoporphyrin IX per cell and had a porphobilinogen deaminase activity of 0.046 fmol uroporphyrin per 30 min per cell. Cells cultured under the same incubation conditions but exposed to 30 mJ cm(-2) irradiation after a 3-h incubation with delta aminolaevulinic acid showed a significant reduction in protoporphyrin IX, 2.28 x 10(-16) mol per cell, and an 80% reduction in porphobilinogen deaminase activity to 0.0088 fmol uroporphyrin per 30 min per cell. Similar effects were evident in irradiated cells incubated with delta-aminolaevulinic acid immediately after, or following a 24 h interval, post-irradiation. There was little gain in efficacy from a second treatment regimen applied within 24 h of the initial treatment, probably a result of initial metabolic damage leading to reduced levels of protoporphyrin IX. These findings suggest that a correlation may exist between the delta-aminolaevulinic acid induction of porphobilinogen deaminase activity and the increase in intracellular protoporphyrin IX accumulation. PMID- 10362108 TI - Enhancement of chemotherapy by manipulation of tumour pH. AB - The extracellular (interstitial) pH (pHe) of solid tumours is significantly more acidic compared to normal tissues. In-vitro, low pH reduces the uptake of weakly basic chemotherapeutic drugs and, hence, reduces their cytotoxicity. This phenomenon has been postulated to contribute to a 'physiological' resistance to weakly basic drugs in vivo. Doxorubicin is a weak base chemotherapeutic agent that is commonly used in combination chemotherapy to clinically treat breast cancers. This report demonstrates that MCF-7 human breast cancer cells in vitro are more susceptible to doxorubicin toxicity at pH 7.4, compared to pH 6.8. Furthermore 31P-magnetic resonance spectroscopy (MRS) has shown that the pHe of MCF-7 human breast cancer xenografts can be effectively and significantly raised with sodium bicarbonate in drinking water. The bicarbonate-induced extracellular alkalinization leads to significant improvements in the therapeutic effectiveness of doxorubicin against MCF-7 xenografts in vivo. Although physiological resistance to weakly basic chemotherapeutics is well-documented in vitro and in theory, these data represent the first in vivo demonstration of this important phenomenon. PMID- 10362109 TI - Amphiregulin acts as an autocrine growth factor in two human polarizing colon cancer lines that exhibit domain selective EGF receptor mitogenesis. AB - Colonic enterocytes, like many epithelial cells in vivo, are polarized with functionally distinct apical and basolateral membrane domains. The aims of this study were to characterize the endogenous epidermal growth factor (EGF)-like ligands expressed in two polarizing colon cancer cell lines, HCA-7 Colony 29 (HCA 7) and Caco-2, and to examine the effects of cell polarity on EGF receptor mediated mitogenesis. HCA-7 and Caco-2 cells were grown on plastic, or as a polarized monolayer on Transwell filters. Cell proliferation was measured by 3H thymidine incorporation and EGF receptor (EGFR) binding was assessed by Scatchard analysis. EGFR ligand expression was determined by Northern blot analysis, reverse transcription polymerase chain reaction, metabolic labelling and confocal microscopy. We found that amphiregulin (AR) was the most abundant EGFR ligand expressed in HCA-7 and Caco-2 cells. AR was localized to the basolateral surface and detected in basolateral-conditioned medium. Basolateral administration of neutralizing AR antibodies significantly reduced basal DNA replication. A single class of high-affinity EGFRs was detected in the basolateral compartment, whereas the apical compartment of polarized cells, and cells cultured on plastic, displayed two classes of receptor affinity. Basolateral administration of transforming growth factor alpha (TGF-alpha) or an EGFR neutralizing antibody also resulted in a dose-dependent stimulation or attenuation, respectively, of DNA replication. However, no mitogenic response was observed when these agents were added to the apical compartment or to confluent cells cultured on plastic. We conclude that amphiregulin acts as an autocrine growth factor in HCA-7 and Caco-2 cells, and EGFR ligand-induced proliferation is influenced by cellular polarity. PMID- 10362110 TI - Antisense oligonucleotides to class III beta-tubulin sensitize drug-resistant cells to Taxol. AB - A major impediment to the successful use of Taxol in the treatment of cancer is the development of drug resistance. The major cellular target of Taxol is the microtubule that is comprised of alpha- and beta-tubulin heterodimers. Binding sites for Taxol have been delineated on the beta-tubulin subunit that has six isotypes. We have recently described increased expression of the brain-specific human class III beta-tubulin isotype, encoded by the Hbeta4 gene, in both Taxol resistant ovarian tumours and non-small-cell lung cancer cell lines. To evaluate directly the role of the class III beta-tubulin isotype in mediating Taxol resistance, antisense phosphorothioate oligodeoxynucleotides (ODN) targeted against various regions of the Hbeta4 gene have been designed and examined for their efficacy in reducing Hbeta4 gene and protein expression. Taxol-resistant lung cancer cells, A549-T24, which are 17-fold resistant to Taxol and display a fourfold increase in Hbeta4 expression compared to the parental A549 cells, were treated with 1 microM antisense ODNs. Two ODNs, AS1 and AS3, were found to reduce mRNA expression by 40-50%, as determined by reverse transcription polymerase chain reaction. A concentration-dependent reduction in Hbeta4 mRNA expression was demonstrated with AS1 ODN. Immunofluorescence staining of cells treated with AS1 ODN revealed a decrease in class III protein expression which corresponded to a 39% increase in sensitivity to Taxol (P < 0.005). These findings support an important role for Hbeta4 (class III) beta-tubulin expression in Taxol resistance and have potential implications for the treatment of Taxol-resistant tumours. PMID- 10362111 TI - [Arg6,D-Trp7,9,NmePhe8]-substance P (6-11) activates JNK and induces apoptosis in small cell lung cancer cells via an oxidant-dependent mechanism. AB - [Arg6,D-Trp7,9,NmePhe8]-substance P (6-11) (antagonist G) is a novel class of anti-cancer agent that inhibits small-cell lung cancer (SCLC) cell growth in vitro and in vivo and is entering phase II clinical investigation for the treatment of SCLC. Although antagonist G blocks SCLC cell growth (IC50 = 24.5 +/- 1.5 and 38.5 +/- 1.5 microM for the H69 and H510 cell lines respectively), its exact mechanism of action is unclear. This study shows that antagonist G stimulates apoptosis as assessed by morphology (EC50 = 5.9 +/- 0.1 and 15.2 +/- 2.7 microM for the H69 and H510 cell lines respectively) and stimulates c-jun-N terminal kinase (JNK) activity in SCLC cells (EC50 = 3.2 +/- 0.1 and 15.2 +/- 2.7 microM). This activity is neuropeptide-independent, but dependent on the generation of reactive oxygen species (ROS) and is inhibited by the free radical scavenger n-acetyl cysteine. Furthermore, antagonist G itself induces inflammation (59% increase in oedema volume compared to control) and potentiates (by 35-40%) bradykinin-induced oedema formation in vivo. In view of these results we show that, as well as acting as a 'broad-spectrum' neuropeptide antagonist, antagonist G stimulates basal G-protein activity in SCLC cell membranes (81 +/- 12% stimulation at 10 microM), thereby displaying a unique ability to stimulate certain signal transduction pathways by activating G-proteins. This novel activity may be instrumental for full anti-cancer activity in SCLC cells and may also account for antagonist G activity in non-neuropeptide-dependent cancers. PMID- 10362112 TI - Magnetic resonance detects metabolic changes associated with chemotherapy-induced apoptosis. AB - Apoptosis was induced by treating L1210 leukaemia cells with mechlorethamine, and SW620 colorectal cells with doxorubicin. The onset and progression of apoptosis were monitored by assessing caspase activation, mitochondrial transmembrane potential, phosphatidylserine externalization, DNA fragmentation and cell morphology. In parallel, 31P magnetic resonance (MR) spectra of cell extracts were recorded. In L1210 cells, caspase activation was detected at 4 h. By 3 h, the MR spectra showed a steady decrease in NTP and NAD, and a significant build up of fructose 1,6-bisphosphate (F-1,6-P) dihydroxyacetonephosphate and glycerol 3-phosphate, indicating modulation of glycolysis. Treatment with iodoacetate also induced a build-up of F-1,6-P, while preincubation with two poly(ADP-ribose) polymerase inhibitors, 3-aminobenzamide and nicotinamide, prevented the drop in NAD and the build-up of glycolytic intermediates. This suggested that our results were due to inhibition of glyceraldehyde-3-phosphate dehydrogenase, possibly as a consequence of NAD depletion following poly(ADP-ribose) polymerase activation. Doxorubicin treatment of the adherent SW620 cells caused cells committed to apoptosis to detach. F-1,6-P was observed in detached cells, but not in treated cells that remained attached. This indicated that our observations were not cell line- or treatment-specific, but were correlated with the appearance of apoptotic cells following drug treatment. The 31P MR spectrum of tumours responding to chemotherapy could be modulated by similar effects. PMID- 10362113 TI - Breast cancer risk in ataxia telangiectasia (AT) heterozygotes: haplotype study in French AT families. AB - Epidemiological studies in ataxia telangiectasia (AT) families have suggested that AT heterozygotes could have an increased cancer risk, especially breast cancer (BC) in women. It has also been suggested that an increased sensibility of AT heterozygotes to the effect of ionizing radiation could be responsible for the increased BC risk. BC relative risk (RR) estimation in AT heterozygotes within families ascertained through AT children is presented here. Family data collected included demographic characteristics, occurrence of cancers, past radiation exposures and blood samples. DNA samples were studied using seven ATM linked microsatellites markers allowing AT haplotypes reconstitution. The relative risk of BC was assessed using French estimated incidence rates. A significant increase risk of BC is found among obligate ATM heterozygotes with a point estimate of 3.32 (P = 0.002). BC relative risk calculated according to age is significantly increased among the obligate ATM heterozygotes female relatives with an age < or = 44 years (RR = 4.55, P = 0.005). The BC relative risk is statistically borderline among the obligate ATM heterozygote female relatives with an age > or = 45 years (RR = 2.48, P = 0.08). The estimated BC relative risk among ATM heterozygotes is consistent with previously published data. However, the increased risk is only a little higher than classical reproductive risk factors and similar to the risk associated with a first-degree relative affected by BC. PMID- 10362114 TI - Cadherin-catenin expression in primary colorectal cancer: a survival analysis. AB - Both cell adhesion and cell signalling events are mediated by components of the cadherin-catenin complex. Loss of expression of the components of this complex have been shown to correlate with invasive behaviour in many tumour types although their exact role in colorectal cancer remains unclear. Immunohistochemical analysis of the expression of components of the cadherin catenin complex in colorectal cancers from 60 patients was undertaken. Loss of memberanous expression of E-cadherin, alpha-catenin and beta-catenin was demonstrated in 52%, 85% and 40% of tumours respectively. Focal nuclear expression of beta-catenin (< 75% of cells per section), usually associated with cytoplasmic expression, was clearly demonstrated in 19 (32%) tumours while widespread nuclear expression (> 75% of tumour cells per section) was seen in 11 (18%) tumours. Loss of membranous alpha-catenin expression significantly correlated with tumour de-differentiation (P = 0.009). There was a trend towards an association between advanced tumour stage and loss of membranous expression of alpha-catenin or beta-catenin, although these associations were not statistically significant. Univariate analysis revealed that advanced Dukes' stage, tumour de differentiation, loss of membranous beta-catenin expression, cytoplasmic beta catenin expression and widespread nuclear expression of beta-catenin all correlated with short survival following apparently curative resection of the primary tumour. However, only Dukes' stage (P = 0.002), tumour grade (P = 0.02) and widespread nuclear expression of beta-catenin (P = 0.002) were independent predictors of short survival. Disturbed growth signalling events in colorectal tumours are thought to result in nuclear accumulation of beta-catenin. Consequently, tumours with widespread nuclear expression of beta-catenin are likely to have severely abnormal growth characteristics, and which therefore might be predictive of short survival in these patients. PMID- 10362115 TI - Phase I study of a biweekly schedule of a fixed dose of cisplatin with increasing doses of paclitaxel in patients with advanced oesophageal cancer. AB - We performed this dose-finding study with a fixed dose of cisplatin and increasing doses of paclitaxel given every 2 weeks to determine the maximum tolerable dose of this schedule. Sixty-four patients with advanced oesophageal cancer were treated with a cisplatin dose of 60 mg m(-2) and increasing doses of paclitaxel from 100 mg m(-2) up to 200 mg m(-2) both administered over 3 h for a maximum of six cycles in patients with stable disease or eight cycles in responding patients. Patients were retreated when the granulocytes were > 0.75 x 10(9) l(-1) and the platelets > 75 x 10(9) l(-1). The dose of paclitaxel could be increased to 200 mg m(-2) without encountering dose limiting haematological toxicity. At the dose levels 190 mg m(-2) and 200 mg m(-2) of paclitaxel cumulative sensory neurotoxicity became the dose-limiting toxicity. The dose intensity of paclitaxel calculated over six cycles rose from 50 mg m(-2) per week to 85 mg m(-2) per week. Only three episodes of granulocytopenic fever were encountered out of a total of 362 cycles of treatment. Of the 59 patients evaluable for response, 31 (52%) had a partial or complete response. In a biweekly schedule with a fixed dose of 60 mg m(-2) cisplatin it is possible to increase the dose of paclitaxel to 180 mg m(-2). At higher dose levels, neurotoxicity becomes the dose-limiting toxicity. The observed response rate warrants further investigation of this schedule. PMID- 10362116 TI - Double-blind randomized study on the myeloprotective effect of melatonin in combination with carboplatin and etoposide in advanced lung cancer. AB - A significant myeloprotective effect of melatonin in mice treated with etoposide, cyclophosphamide or carboplatin has been reported. The present study was designed to evaluate if the same effect could be observed in patients receiving chemotherapy. Twenty previously untreated patients with inoperable lung cancer received two cycles of carboplatin (given at area under the curve 5 by the Calvert formula) on day 1 and etoposide (150 mg m(-2) i.v.) on days 1-3 every 4 weeks. Melatonin 40 mg or placebo (double-blind) was given orally in the evening for 21 consecutive days, starting 2 days before chemotherapy. Patients were randomized to receive melatonin either with the first or the second cycle. Complete blood cell count with differential was done three times per week for 3 weeks. The median age of the cohort was 60 years (range 42-69), 16 patients had non-small cell and four patients small-cell lung cancer, 12 stage III and eight stage IV disease. In a multivariate analysis including age, sex, diagnosis, stage, performance status, doses of carboplatin and etoposide, and concomitant treatment with melatonin or placebo, the haematological parameters--depth and duration of toxicity for haemoglobin, platelets and neutrophils (ANC)--were not significantly different between cycles with/without melatonin. The mean ANC nadir and the mean number of days with ANC < 0.5 x 10(9) l(-1) were 0.5 x 10(9) l(-1) and 2.5 days, respectively, with/without melatonin. We concluded that, in patients with lung cancer, melatonin given orally at a dose of 40 mg per day for 21 days in the evening, does not protect against the myelotoxic effect of carboplatin and etoposide. PMID- 10362117 TI - Growth dysregulation and p53 accumulation in human primary colorectal cancer. AB - p53 accumulation is common in colorectal cancer, but effects on growth homeostasis are unclear. In this study, DNA content, cell cycle phase fractions and DNA strand-breaks consistent with apoptosis were assessed by flow cytometry in 42 fresh primary colorectal tumours and matched normal mucosa. p53 accumulation was assessed in 37 fixed tumour sections, by immunohistochemistry. In normal mucosa, 10.3 +/- 6.6% (mean +/- s.d.) cells were in DNA synthesis phase while 28.7 +/- 17.9% showed apoptosis. A relationship suggestive of growth homeostasis, was observed between these parameters (r = 0.8; P < 0.05). In cancers, a greater number of cells were in DNA synthesis phase (15.6 +/- 12.9% tumour vs mucosa 10.3 +/- 6.6%; P < 0.02) while fewer showed apoptosis than normal mucosa (18.5 +/- 17.0% tumour vs mucosa 28.7 +/- 17.9%; P < 0.01). DNA synthesis and apoptosis fractions were unrelated in cancers, suggesting growth dysequilibrium. p53 accumulation was detected in 59% (22/37) tumours and was associated with reduced apoptosis compared to p53-negative tumours or mucosa (14.8 +/- 15% p53 accumulation vs 26.3 +/- 18% p53-negative; P < 0.05; vs 28.7 +/ 17.9% mucosa; P < 0.05). p53 accumulation was unrelated to DNA synthesis phase fractions. p53 accumulation is accompanied by reduced apoptosis which may accentuate growth dysequilibrium in colorectal cancer. PMID- 10362118 TI - Expression of p73, a novel protein related to the p53 tumour suppressor p53, and apoptosis in cholangiocellular carcinoma of the liver. AB - p73, the first homologue of the tumour suppressor protein p53, was recently discovered on chromosome 1p36 and has been shown to induce apoptosis in a p53 like manner. The present study was performed with the aim of investigating the expression of p53, its new homologue p73 and the occurrence of apoptosis in cholangiocellular carcinoma. Protein levels of p73 were examined in 41 patients with curatively (R0-) resected cholangiocellular carcinomas with an antiserum, raised against a peptide in the N-terminal domain of p73. The incidence of mutations in the p53 gene was analysed by direct sequencing and also immunohistochemically. Apoptotic cell death was assessed using in-situ end labelling (ISEL) technique in combination with morphological criteria. The results obtained were correlated with patient survival. Immunostaining of p73 protein was detected in 17/41 carcinomas examined (41%). The immunoreactivity was confined to the cell nucleus. In 15/41 patients (37%), mutations of the p53 gene were observed. Eleven out of these 15 patients stained also positive for p73. In contrast, out of 26 patients without any detectable p53 mutation, only six exhibited p73 immunostaining. We failed to observe a correlation between p73 expression or p53 and apoptosis within a given tumour. Survival analysis including the parameters stage and grade of disease, p73 and p53, and also apoptosis, showed that tumour stage and grade as well as p53 and p73 were significantly related to prognosis. In Cox regression survival analysis, however, only extent of primary tumour and lymph node status had an independent prognostic impact. Our results with a high prevalence of p73 within tumours harbouring mutated p53 gene suggest that p73 could compensate for p53 function. We failed to establish p73 or p53 as independent prognostic factors in cholangiocellular carcinoma of the liver. PMID- 10362119 TI - Bcl-2 expression related to altered p53 protein and its impact on the progression of human pancreatic carcinoma. AB - p53 and Bcl-2 are two important factors related to apoptosis and tumorigenesis. In this study, a series of 52 cases of pancreatic carcinoma (PC) were investigated using an immunohistochemical assay to determine whether altered expression of Bcl-2 and p53 has an impact on the progression of this malignancy. Cytoplasmic immunoreactivity for Bcl-2 and nuclear staining of p53 was found in 12 (23.1%) and 32 (63.5%) cases of PC respectively. Furthermore, an inverse correlation between the expression of p53 and Bcl-2 existed in this series (P < 0.01). In a subgroup, the proportion of tumours showing that p53-positive and Bcl 2-negative staining was increased with increasing histological grade and clinical stage (P < 0.05), and moreover, the survival period of those patients whose tumour had this staining was shorter than those with other staining patterns of combined p53 and Bcl-2 (P < 0.05). Therefore, it is concluded that simultaneously aberrant expression of Bcl-2 and p53 may confer PC with more malignant clinicopathological characteristics. PMID- 10362120 TI - Mutational spectrum of p53 gene in arsenic-related skin cancers from the blackfoot disease endemic area of Taiwan. AB - To understand the role of p53 tumour suppressor gene in the carcinogenesis of arsenic-related skin cancers from the blackfoot disease endemic area of Taiwan, we collected tumour samples from 23 patients with Bowen's disease, seven patients with basal cell carcinomas (BCC) and nine patients with squamous cell carcinomas (SCC). The result showed that p53 gene mutations were found in 39% of cases with Bowen's disease (9/23), 28.6% of cases with BCC (2/7) and 55.6% of cases with SCC (5/9). Most of the mutation sites were located on exon 5 and exon 8. Moreover, the results from direct sequencing indicated that missense mutations were found at codon 149 (C-->T) in one case, codon 175 (G-->A) in three cases, codon 273 (G- >C) in three cases, codon 292 (T-->A) in one case, codon 283 (G-->T) in one case, codon 172 (T-->C) in one case and codon 284 (C-->A) in one case. In addition, silent mutations were also found in four cases. These mutations were located at codons 174, 253, 289 and 298 respectively. In immunohistochemistry analysis, p53 overexpression was found in 43.5% (10/23) of cases with Bowen's disease, 14% (1/7) of cases with BCC and 44% (4/9) of cases with SSC. These findings showed that p53 gene mutation rate in arsenic-related skin cancers from the blackfoot disease endemic area of Taiwan is high and that the mutation types are different from those in UV-induced skin cancers. PMID- 10362121 TI - Expression of matrix metalloproteinase-1, -2 and -3 in squamous cell carcinoma and actinic keratosis. AB - Matrix metalloproteinase (MMP) plays an important role in extracellular matrix degradation associated with cancer invasion. An expression of MMP-1 (interstitial collagenase), MMP-2 (72-kDa type IV collagenase) and MMP-3 (stromelysin-1) was investigated in squamous cell carcinoma (SCC) and its precancerous condition, actinic keratosis (AK), using in situ hybridization techniques. MMP-1 mRNA was detected in tumour cells and/or in stromal cells in all cases of SCC, four of six AKs adjacent to SCC and four of 16 AKs. MMP-2 and MMP-3 mRNAs were detected in SCC but not in AK. The expression of MMP-3 correlated to that of MMP-1 (P = 0.03) localized at the tumour mass and stroma of the invasive area, while MMP-2 mRNA was detected widely throughout the stroma independent of MMP-1 expression. Our results indicated that the expression of MMP-1, -2 and -3 showed different localization patterns, suggesting a unique role of each MMP in tumour progression. Moreover, MMP-1 expression could be an early event in the development of SCC, and AK demonstrating MMP-1 mRNA, might be in a more advanced dysplastic state, progressing to SCC. PMID- 10362122 TI - Exposure to female hormone drugs during pregnancy: effect on malformations and cancer. AB - This study aimed to investigate whether the use of female sex hormone drugs during pregnancy is a risk factor for subsequent breast and other oestrogen dependent cancers among mothers and their children and for genital malformations in the children. A retrospective cohort of 2052 hormone-drug exposed mothers, 2038 control mothers and their 4130 infants was collected from maternity centres in Helsinki from 1954 to 1963. Cancer cases were searched for in national registers through record linkage. Exposures were examined by the type of the drug (oestrogen, progestin only) and by timing (early in pregnancy, only late in pregnancy). There were no statistically significant differences between the groups with regard to mothers' cancer, either in total or in specified hormone dependent cancers. The total number of malformations recorded, as well as malformations of the genitals in male infants, were higher among exposed children. The number of cancers among the offspring was small and none of the differences between groups were statistically significant. The study supports the hypothesis that oestrogen or progestin drug therapy during pregnancy causes malformations among children who were exposed in utero but does not support the hypothesis that it causes cancer later in life in the mother; the power to study cancers in offspring, however, was very low. Non-existence of the risk, negative confounding, weak exposure or low study-power may explain the negative findings. PMID- 10362123 TI - Risk factors for testicular cancer: a case-control study in twins. AB - Early life and anthropometric risk factors for testicular cancer were examined in a case-control study in England and Wales in which affected male twins were compared with their unaffected male co-twins. Questionnaire data was obtained for 60 twin pairs. Significantly raised risk of testicular cancer occurred in twins who had longer arms and legs than their co-twin. There was a significant excess of testicular cancer reported in non-twin brothers, as well as in twin brothers, of cases. Risk was also significantly raised in relation to cryptorchidism. The results on limb length suggest that factors, perhaps nutritional, affecting growth before puberty, may be causes of testicular cancer. The results on risk in brothers add to evidence of a large genetic component in aetiology of the tumour. The risk associated with cryptorchidism in the twins accords with the hypothesis that cryptorchidism is causally associated with testicular cancer because it is a cause of the malignancy, rather than because the same maternal factors experienced in utero cause both conditions. PMID- 10362124 TI - Human herpes virus-6 seroprevalence and leukaemias: a case-control study. GIMEMA (Gruppo Italiano Malattie Ematologiche dell' Adulto). AB - The relationships between acute myeloid leukaemia (AML), acute lymphocytic leukaemia (ALL), chronic myeloid leukaemia (CML) and refractory anaemia with excess of blasts (RAEB) and human herpes virus (HHV)-6 antibody level were investigated in a multicentre case-control study. An association between increased HHV-6 seropositivity and geometric mean titre ratio with AML was shown: P for trend = 0.022, adjusted odds ratio 1.20, 95% confidence interval 1.07-1.33 respectively. No association was found between HHV-6 and ALL, CML or RAEB. PMID- 10362126 TI - Progestins and the endometrium in patients receiving tamoxifen. PMID- 10362125 TI - Animal products, calcium and protein and prostate cancer risk in The Netherlands Cohort Study. AB - Prostate cancer risk in relation to consumption of animal products, and intake of calcium and protein was investigated in the Netherlands Cohort Study. At baseline in 1986, 58,279 men aged 55-69 years completed a self-administered 150-item food frequency questionnaire and a questionnaire on other risk factors for cancer. After 6.3 years of follow-up, 642 prostate cancer cases were available for analysis. In multivariate case-cohort analyses adjusted for age, family history of prostate cancer and socioeconomic status, no associations were found for consumption of fresh meat, fish, cheese and eggs. Positive trends in risk were found for consumption of cured meat and milk products (P-values 0.04 and 0.02 respectively). For calcium and protein intake, no associations were observed. The hypothesis that dietary factors might be more strongly related to advanced prostate tumours could not be confirmed in our study. We conclude that, in this study, animal products are not strongly related to prostate cancer risk. PMID- 10362127 TI - Vascular endothelial growth factor is up-regulated in the early pre-malignant stage of colorectal tumour progression. AB - Angiogenesis is an essential requirement for the development, progression and metastasis of malignant tumours. Studies on transgenic mouse models have shown that angiogenesis begins in the pre-malignant phase of oncogenesis, when dysplastic lesions acquire an increased microvasculature. To investigate the relationship between the expression of vascular endothelial growth factor (VEGF) and colorectal tumour progression, we have studied VEGF expression level and splice variant pattern by semi-quantitative RT-PCR and the cellular source of VEGF expression by in situ hybrization (ISH) in a range of lesions that modelled the tumour-development pathway from normal colon to invasive colorectal adenocarcinomas. Colonic adenomas showed a statistically significant up regulation of VEGF expression over normal tissues, with a further increase during the development of adenocarcinomas. Tumour cells formed the major source of VEGF expression, with a minor contribution from mononuclear cells in the tumour stroma and enhanced expression in tumour cells around necrotic regions. The comparable expression level in both the in situ and invasive components in the same tumours indicated that a high VEGF expression capacity had been acquired prior to establishment of the invasive phenotype. Our findings support activation of VEGF as the molecular basis for the discrete induction of angiogenesis in the pre malignant phase of colorectal tumour development. PMID- 10362129 TI - Fine deletion mapping of chromosome 8p in non-small-cell lung carcinoma. AB - Several somatic genetic alterations have been described in non-small-cell lung carcinomas (NSCLC). Recurrent chromosomal deletions have suggested the presence of tumor-suppressor genes specifically involved in lung carcinogenesis. For one of these, 2 non-overlapping regions have been proposed on the short arm of chromosome 8, encompassing the LPL and NEFL genes. The LPL region has been extensively studied in NSCLC and other cancer types. Two genes, N33 and PRLTS, have been identified, but the small number of mutations excludes their involvement in the vast majority of tumors. In order to delineate a reliable region of deletional overlap on chromosome 8p in NSCLC, a series of 77 NSCLC was studied for 34 microsatellite polymorphisms distributed on chromosome 8p, using multiplex-PCR amplification. After purification of tumor nuclei by flow cytometry based on either the abnormal DNA index or the presence of a high expression of cytokeratin, allelic losses on chromosome 8p were observed in 39% of cases. Measurement of DNA index showed that 62% of tumors were hyperploid; allelic losses were more frequent in hyperploid than in diploid tumors (54% vs. 14%; p < 10(-4)). Deletions of part of the short arm were observed in 7 instances. Our data allow definition of an interval of common deletion, flanked by the loci D8S511 and D8S1992, where the putative tumor-suppressor gene might be localized. PMID- 10362128 TI - Gene mutation of transforming growth factor beta1 type II receptor in hepatocellular carcinoma. AB - Alteration of transforming growth factor beta1 (TGF-beta1) type II receptor (RII) appears to cause unresponsiveness to TGF-beta1 in tumorigenic cells. Defect in the mononucleotide repeat sequence, i.e., poly A region of TGF-beta1RII gene has been reported to be related to replication error-positive cancer cells. We examined if there is any TGF-beta1RII mutation in a coding microsatellite in hepatocellular carcinoma (HCC). Genomic DNAs were extracted from formalin-fixed, paraffin-embedded liver tissues obtained at surgery or autopsy in 3 normal individuals and 96 patients with hepatitis C virus-induced chronic liver disease; 3 with chronic hepatitis, 20 with liver cirrhosis and 73 with HCC. The DNA was PCR-amplified at 2 segments of TGF-beta1RII: poly A region which includes the (A)10 microsatellite sequence, and poly GT region. PCR products were directly sequenced. DNA from normal and patients with chronic liver disease contained the 10 wild-type adenines but 3 cases with liver cirrhosis in whom there were only 9 adenines within poly A tract. This microdeletion of one A resulted in a frameshift and truncated a predicted length of amino acids. In HCC lesions, the same deletion was noted in 4 cases (25%) of well-differentiated type, 10 (40%) of moderately differentiated type, 18 (53%) of poorly differentiated type. None of the lesions had mutations within the GT region. Our findings indicate that one adenine deletion of poly A microsatellite tract within TGF-beta1RII is frequently detected in patients with HCC, and the mutation may cause the abrogation of the function of TGF-beta1RII gene. PMID- 10362131 TI - Immunophenotypic and genotypic characterization of nasal lymphoma with polymorphic reticulosis morphology. AB - Nasal lymphoma with polymorphic reticulosis (PR) morphology is now categorized as T/natural killer (T/NK) cell lymphoma. In this study, immunophenotypes and genotypes of proliferating cells in 21 cases with PR were examined. The patients included 13 men and 8 women ranging in age from 20 to 74 (median 37) years. All patients presented with lesions in the upper respiratory tract, mostly in the nasal cavity. Histological specimens obtained from the primary lesions (19 cases) and metastatic cervical lymph nodes (2 cases) were used for analyses. Histologically, polymorphous proliferation was found in 20 cases, and these were thus diagnosed as PR. A monomorphous pattern was found in the remaining last case. Immunohistochemical analysis revealed that the proliferating cells were CD56 (123C3)+ and/or CD16 (2H7)+, TIA-1+ and frequently stained CD3 epsilon+. Tumor cells were frequently stained positively with monoclonal antibodies (mAbs) for T lymphocytes, but were negative for T-cell receptor (TCR) beta and delta chain expression. In situ hybridization analysis using an Epstein-Barr virus encoded early RNA 1 (EBER-1) probe revealed positive signals in 13 of the 15 cases examined. Southern blotting analysis for clonality of the Epstein-Barr virus (EBV) genome in 12 positive cases confirmed the presence of monoclonal proliferation in 7 cases. The pattern of TCR gamma chain gene rearrangement was examined by PCR analysis of DNA from tumor tissues by the denaturing gradient gel electrophoresis method. The results demonstrated no clonal rearrangement in any of the 21 cases examined, including 7 cases with proven clonal proliferation of EBV-infected cells, indicating the absence of T-cell clones. Our findings strongly suggested that nasal T-cell lymphoma is in fact a NK cell lymphoma. PMID- 10362130 TI - Rapid quantification of HTLV-I provirus load: detection of monoclonal proliferation of HTLV-I-infected cells among blood donors. AB - In this report, we quantified HTLV-I provirus load using the AmpliSensor system, which utilizes fluorescence to measure PCR products. With this method, provirus loads could be measured within 6 hr, and the results obtained correlated well with those obtained by other methods. Samples from 256 blood donors, who were positive for antibodies against HTLV-I, were analyzed, showing that provirus load ranged from less than 0.1% to 56% among carriers. We analyzed the association between provirus load and the biomarkers age and sex and found that it was not influenced by either. Provirus load was better correlated with soluble interleukin-2 receptor (sIL-2R) levels than with antibody titer against the virus. Among 18 blood donors with high provirus load (more than 10%), Southern blotting detected monoclonal integration of HTLV-I in infected cells in 2 cases, both of them showing high sIL-2R levels (more than 900 U/ml). Sequential analyses of provirus load showed stable levels of provirus in the same carriers, suggesting that some factors other than age or sex determined provirus load in infected individuals. Thus, this rapid method is a useful tool for the early detection of adult T-cell leukemia and other HTLV-I-associated diseases. PMID- 10362132 TI - Risk of stomach cancer in relation to consumption of cigarettes, alcohol, tea and coffee in Warsaw, Poland. AB - To identify reasons for the high incidence rates of stomach cancer in Poland, we conducted a population-based case-control study in Warsaw. Cases were residents aged 21 to 79 years who were newly diagnosed with stomach cancer between March 1, 1994, and April 30, 1997. Controls were randomly selected from Warsaw residents registered at the nationwide Polish Electronic System of Residence Evidency, frequency-matched to cases by age and sex. Information on demographic characteristics; consumption of cigarettes, alcohol, tea and coffee; diet; medical history; family history of cancer; occupational history; and living conditions during adolescence was elicited by trained interviewers using a structured questionnaire. Included were 464 cases (90% of eligible) and 480 controls (87% of eligible). Among men, the risk of stomach cancer was significantly elevated among current smokers (OR = 1.7, 95% CI = 1.1-2.7) but not among former smokers. The excess risk was largely confined to long-term and heavy smokers, with significant 2-fold excess risk among men who smoked 40 or more pack years. Among women, an 80% increase in risk was observed in both current and former smokers but dose-response trends were less consistent than among men. Alcohol consumption was not clearly related to risk, and no association was found for drinking regular coffee or herbal tea or using milk/cream in coffee or tea. A significant reduction in risk was linked to daily tea drinking among women, but not among men. Our findings confirm an association with cigarette smoking, which is estimated to account for approximately 20% of stomach cancers diagnosed among Warsaw residents during the study period. PMID- 10362133 TI - Breast cancer in New South Wales in 1972-1995: tumor size and the impact of mammographic screening. AB - To examine the use of mammographic screening in women in New South Wales (NSW), we measured uptake of initial mammograms and estimated the proportions of breast cancers that were screen detected. To see if mammographic screening has been associated with reductions in advanced breast cancers and mortality from breast cancer, we analyzed trends in age-specific and age-standardized breast cancer incidence and mortality from 1972 to 1995 and tumor size in 1986, 1989, 1992 and April to September 1995. Between 1984 and the end of 1995, an estimated 72% of NSW women in their 50s and 67% in their 60s had had at least 1 mammogram and, in 1995, an estimated 39% of invasive breast cancers in women in these age groups were detected by mammography. Before 1989, breast cancer incidence increased only slightly (+1.3% annually) but then, from 1990 to 1995, increased more rapidly (+3.1% annually). Between 1986 and 1995, rates of small cancers (< 1 cm) increased steeply by 2.7 times in women 40-49 years of age and 5.6 times in women 50-69 years of age. The incidence of large breast cancers (3+ cm), after little apparent change to 1992, fell by 17% in women 40-49 years of age and 20% in those 50-69 years of age to 1995. Breast cancer mortality increased slightly between 1972 and 1989 (+0.5% annually) but then fell (-2.3% annually) from 1990 to 1995. We concluded that breast cancer rates had been influenced in expected directions by the introduction of mammographic screening in women resident in NSW. We expect that recent falls in incidence of larger breast cancers and breast cancer mortality will become steeper as screening coverage increases in the second half of the 1990s. PMID- 10362134 TI - Immune response to human papillomavirus 16 L1E7 chimeric virus-like particles: induction of cytotoxic T cells and specific tumor protection. AB - Expression of human papillomavirus type 16 (HPV 16) fusion proteins LI deltaCE7(1 55) and LI deltaCE7(1-60) (carboxy-terminal deletion of LI replaced by 55 or 60 amino-terminal amino acids of E7) leads to formation of chimeric papillomavirus like particles (CVLPs). After "infection" of cells by CVLPs, the chimeric proteins can be detected in the cytosol and the endoplasmic reticulum (ER), suggesting that they are intracellularly processed via the MHC class I pathway and, therefore, able to activate cytotoxic T lymphocytes (CTLs). To investigate the cytotoxic immune response against HPV 16 LI deltaCE7(1-60) and LI deltaCE7(1 55) CVLPs, we immunized C57Bl/6 mice with various CVLP doses without adjuvant. Two weeks after immunization, spleen cells were prepared and stimulated in vitro using HPV 16 E7-expressing transfectants of the tumor cell line RMA. In 51Cr release cytotoxicity assays, spleen cells of mice vaccinated with LI deltaCE7(1 60) CVLPs specifically lysed the RMA-E7 transfectants as well as RMA cells loaded with the peptide E7(49-57), which represents an H2-Db-restricted CTL epitope. This demonstrates that CVLPs induce an E7-specific CTL response in mice in the absence of an adjuvant. Furthermore, immunization with CVLPs prevented outgrowth of E7-expressing tumor cells even if inoculation of cells was performed 2 weeks before vaccination. We conclude from our data that CVLPs show promise for therapy of HPV-associated lesions. PMID- 10362135 TI - Nitric-oxide production by murine mammary adenocarcinoma cells promotes tumor cell invasiveness. AB - The role of nitric oxide (NO) in tumor biology remains controversial and poorly understood. While a few reports indicate that the presence of NO in tumor cells or their micro-environment is detrimental for tumor-cell survival, and consequently their metastatic ability, a large body of data suggests that NO promotes tumor progression. The purpose of this study was to identify the source of NO in the spontaneously metastasizing C3-L5 murine mammary-adenocarcinoma model, the role of tumor-derived NO in tumor-cell invasiveness, and the mechanisms underlying the invasion-stimulating effects of tumor-derived NO. The source of NO was established by immunocytochemical localization of NO synthase (NOS) enzymes in C3-L5 cells in vitro and transplanted tumors in vivo. An in vitro transwell Matrigel invasion assay was used to test the invasiveness of C3 L5 cells in the presence or the absence of NO blocking agents or iNOS inducers (IFN-gamma and LPS). The mechanisms underlying the invasion-stimulating effects of tumor-derived NO were examined by measuring mRNA expression of matrix metalloproteinases (MMP)-2 and -9, and tissue inhibitors of metalloproteinases (TIMP) 1, 2 and 3 in C3-L5 cells in various experimental conditions. Results showed that C3-L5 cells expressed high level of eNOS protein in vitro, and in vivo, both in primary and in metastatic tumors. C3-L5 cells also expressed iNOS mRNA and protein when cultured in the presence of IFN-gamma and LPS. Constitutively produced NO promoted tumor-cell invasiveness in vitro by down regulating TIMP 2 and TIMP 3. In addition, there was up-regulation of MMP-2, when extra NO was induced by IFN-gamma and LPS. In conclusion, NO produced by C3-L5 cells promoted tumor-cell invasiveness by altering the balance between MMP-2 and its inhibitors TIMP-2 and 3. Thus, our earlier observations of anti-tumor and anti-metastatic effects of NO inhibitors in vivo in this tumor model can be explained, at least in part, by reduced tumor-cell invasiveness. PMID- 10362136 TI - Pattern of FHIT gene expression in normal and leukaemic cells. AB - Chromosomal aberrations and inactivation of tumour suppressor genes are frequent in acute leukaemia. To determine whether the FHIT gene is involved in the development of leukaemia, we examined the FHIT transcript in 65 leukaemia cell lines, 5 fresh acute leukaemia patients at diagnosis and in complete remission, normal peripheral blood lymphocytes obtained from 14 healthy volunteers and Epstein-Barr (EB) virus transformed 5 B-cell lines (EB-lines), using nested reverse transcription-polymerase chain reaction and direct sequencing. The transcripts were classified into 4 patterns: pattern I revealed the normal transcripts only, pattern II the altered transcripts in addition to the normal transcripts, pattern III the altered transcripts without the normal transcripts and pattern IV an absence of normal and altered FHIT transcripts. Nineteen cell lines were classified as pattern I, 32 as pattern II, 2 as pattern III and 12 as pattern IV. The frequency of loss of FHIT expression (pattern III or IV) varied in each type of leukaemia cell line; the order ranked from the highest incidence was acute myeloid leukaemia (AML), T-cell acute lymphoblastic leukaemia (T-ALL), B-precursor ALL, B-ALL, and chronic myeloid leukaemia (CML). No genomic rearrangement was found in any samples examined. All of 5 patients showed same pattern II FHIT transcripts at 2 different stages of the disease. All normal peripheral blood lymphocytes and EB-lines were classified as pattern I or II. Our results suggested that patterns III and IV of FHIT transcripts might be associated with the development of a subset of leukaemia, while pattern II which has so far been reported as an aberrant transcript in varieties of malignant tumours might not be associated with leukaemogenesis. PMID- 10362137 TI - Establishment and characterization of 12 human colorectal-carcinoma cell lines. AB - In this article, we describe the characteristics of 12 human colorectal-carcinoma cell lines established from 6 primary tumors and 6 metastatic sites of 11 Korean colorectal-carcinoma patients, including the morphology in vivo and in vitro and mutations of K-ras2, p15, p16, p53, APC, beta-catenin, hMLH1 and hMSH2 genes in vitro. No lines were contaminated with Mycoplasma or bacteria. All lines were proven to be unique by DNA-fingerprinting analysis. All lines expressed the surface carcino-embryonic antigen and secreted it into the supernatant fluid. The morphological correlation between the original tumors and cultured cells suggested that the original tumors showing mucinous adenocarcinoma correlated with floating aggregates in culture, and degree of desmoplasia in the original tumor correlated with attached growth in culture. Five of the cell lines showed mutations in the K-ras2 gene, and 6 of the cell lines showed mutations in the p53 gene. The p15 gene was deleted in 2 cell lines, and the p16 gene was hypermethylated in 3 cell lines. The mutation of mismatch-repair genes (hMLH1 and hMSH2) was found in 4 lines, the APC gene and beta-catenin gene were mutated in 9 and 2 lines respectively. These well-characterized colorectal-cancer cell lines should serve as useful tools for investigating the biological characteristics of colorectal cancer. PMID- 10362138 TI - Binding characteristics and tumor targeting of a covalently linked divalent CC49 single-chain antibody. AB - Multivalency is a recognized means of increasing the functional affinity of single-chain Fvs (scFvs) for optimizing tumor uptake. A unique divalent single chain Fv protein [sc(Fv)2], based on the variable regions of the monoclonal antibody (MAb) CC49, has been generated that differs from other dimeric single chain constructs in that a linker sequence (L) is encoded between the repeated V(L) and V(H) domains (V(L)-L-V(H)-L-V(L)-L-V(H)). This construct was expressed in soluble form in Escherichia coli and purified by ion-exchange and gel filtration chromatography. Purity and immunoreactivity were determined by SDS PAGE, HPLC and competitive RIA. sc(Fv)2 exhibited a relative K(A) (3.34 x 10(7) M(-1)) similar to that of the native IgG (1.14 x 10(8) M(-1)) as determined by BIAcore analysis. Pharmacokinetic studies showed rapid blood clearance for sc(Fv)2, with a T(1/2) less than 40 min. Whole-body clearance analysis also revealed rapid clearance, suggesting no significant retention in the extravascular space or normal tissues. Biodistribution studies of radiolabeled sc(Fv)2 showed tumor uptake greater than 6% ID/g after 30 min, which remained at this level for 6 hr. High tumor uptake and retention of sc(Fv)2 coupled with rapid blood and whole-body clearance makes this dimeric scFv of MAb CC49 a strong candidate for imaging and therapeutic applications. PMID- 10362139 TI - Inhibition of tumor invasion and metastasis by a novel lysophosphatidic acid (cyclic LPA). AB - Fetal calf serum (FCS) and 1-oleoyl lysophosphatidic acid (LPA) were previously found to be potent inducers of invasion (transcellular migration) in an in vitro system. A novel LPA, composed of cyclic phosphate and cyclopropane-containing hexadecanoic acid (PHYLPA), first isolated from myxoamoebae of Physarum polycephalum, and its synthetic derivatives (cLPA) were tested for their ability to inhibit tumor cell invasion and metastasis. Amoung these, Pal-cLPA, which has a palmitoyl moiety, was most potent in inhibiting invasion, with 93.8% inhibition at the concentration of 25 microM. Invasion in vitro by mouse melanoma cells (B16), human pancreatic adenocarcinoma cells (PSN-1), human lung cancer cells (OC 10) and human fibrosarcoma cells (HT-1080) was also inhibited by Pal-cLPA. The stimulation of MMI cells with LPA triggered F-actin formation, which was impaired by the addition of Pal-cLPA at invasion-inhibitory concentration. Pal-cLPA induced a rapid increase in adenosine 3',5'-cyclic monophosphate (cAMP) concentration in MMI cells. The addition of dibutyryl cAMP significantly abrogated LPA-induced invasion by MM1 cells and actin polymerization in the cells. The inhibition of MM1 cell invasion by Pal-cLPA may be ascribed to an increased level of cAMP. Pal-cLPA also suppressed invasion in vitro by MM1 cells induced by FCS dose dependently, without affecting proliferation. It also suppressed the pulmonary metastasis of B16 mouse melanoma cells injected into the tail vein of C57BL/6 mice. Thus, Pal-cLPA is effective in inhibiting invasion and metastasis of a variety of tumor cells. PMID- 10362140 TI - Berberine-induced apoptosis of human leukemia HL-60 cells is associated with down regulation of nucleophosmin/B23 and telomerase activity. AB - The steady-state level of nucleophosmin/B23 mRNA decreased during berberine induced (25 microg/ml, 24 to 96 hr) apoptosis of human leukemia HL-60 cells. A decline in telomerase activity was also observed in HL-60 cells treated with berberine. A stable clone of nucleophosmin/B23 overexpressed in HL-60 cells was selected and found to be less responsive to berberine-induced apoptosis. About 35% to 63% of control vector-transfected cells (pCR3) exhibited morphological characteristics of apoptosis, while about 8% to 45% of nucleophosmin/B23-over expressed cells (pCR3-B23) became apoptotic after incubation with 15 microg/ml berberine for 48 to 96 hr. DNA extracted from pCR3 cells contained more fragmented DNA than pCR3-B23 cells during treatment with 15 microg/ml berberine for 24 to 48 hr. Our results indicate that berberine-induced apoptosis is associated with down-regulation of nucleophosmin/B23 and telomerase activity. We also suggest that nucleophosmin/B23 may play an important role in the control of the cellular response to apoptosis induction. PMID- 10362141 TI - Concurrent deregulation of gelsolin and cyclin D1 in the majority of human and rodent breast cancers. AB - Decreased gelsolin and increased cyclin D1 are among the most common defects found in human and rodent breast cancers. Our purpose was to determine the frequency of concurrence of these 2 alterations in this malignancy. Our results demonstrate that gelsolin protein and mRNA were significantly reduced in 80-100% of rodent mammary carcinomas that developed spontaneously, following oncogene introduction, or after treatment with viral, chemical or hormonal agents. The reduction in gelsolin most likely occurs during the transition from preneoplasia to carcinoma because hyperplasias had normal levels of gelsolin whereas microtumors had reduced expression. Southern analysis revealed no major mutations in the gelsolin gene of tumors with low expression. Cyclin D1 mRNA was increased in 50-100% of these rodent mammary tumors, although the cyclin D1 gene was not amplified. By nuclear runon assay, downregulation of gelsolin in both human and mouse mammary cancer cells involved diminished transcription and, conversely, human breast cancer cells expressing high levels of cyclin D1 had increased initiation of cyclin D1 transcription compared with cyclin D1 low expressors. Thus, alteration in the rate of transcription appears to be an important factor underlying the dysfunction of these genes. According to our data, concurrent deregulation of gelsolin and cyclin D1 is highly prevalent among breast cancers of humans and rodents, with both defects present in 89% of the neoplasms analyzed in this study. In fact, most tumors in every rodent model of mammary tumorigenesis examined had the 2 alterations. PMID- 10362142 TI - Expression of cytokeratin 19 mRNA in human lung cancer cell lines. AB - The present study was designed to clarify the mechanism by which some lung cancer cell lines can produce cytokeratin 19 (CK19) fragment and others cannot. We hypothesized that some lung cancer cell lines which cannot release CK19 express an incomplete sequence of CK19 mRNA. Expression of mRNA was evaluated by RT-PCR using several primer pairs for CK19. CK19 in the culture supernatant was measured by an immuno-radiometric assay. CK19 protein synthesis was evaluated by Western immunoblot and immunohistochemistry. Among 16 lung cancer cell lines, 7 released significant amounts of CK19 in the supernatant. In some cell lines, expression of CK19 mRNA was observed only in some combinations of primers, suggesting that incomplete mRNA was expressed. 3'-RACE analysis detected amplified products of a shorter size compared with normal amplified products in cell lines which expressed incomplete CK19 mRNA, suggesting that 3'-ends of mRNA for CK19 were deleted. Results of Western immunoblot and immuno-histochemical staining using anti-human CK19 monoclonal antibody completely correlated with the results on CK19 levels in culture supernatants as well as with complete expression of mRNA. We conclude that levels of CK19 closely relate to the expression of complete mRNA for CK19. PMID- 10362143 TI - Retrovirus-mediated transfer of anti-MDR1 hammerhead ribozymes into multidrug resistant human leukemia cells: screening for effective target sites. AB - One of the underlying mechanisms of multidrug resistance (MDR) is cellular over production of P-glycoprotein (P-gp), which acts as a drug efflux pump. P-gp is encoded by a small group of related genes termed MDR; only MDR1 is known to confer drug resistance. To overcome P-gp-mediated drug resistance, we have developed two anti-MDR1 hammerhead ribozymes driven by the beta-actin promoter. Upon transduction of the ribozymes into MDR cells, vincristine resistance was decreased. These two ribozymes were constructed, which showed different cleavage activities. In this study, to determine suitable target sites for the anti-MDR1 ribozyme, the exon 1b-intron 1 boundary, the translation-initiation site, the intron 1-exon 2 boundary and the exon 2-intron 2 boundary, codons 179 and 196 of the MDR1 gene were selected as candidates. To improve the ribozyme activity, a retroviral vector containing RNA polymerase III promoter was used. Stable retrovirus producer cells were generated by transfecting the retroviral vector plasmids carrying the ribozyme into the packaging cell line. Retroviral vector transduction of human leukemia cell lines expressing MDR1 was accomplished by co culturing these with virus producer cells. Stably transduced cells were selected by G418 and pooled to determine the efficacy of each ribozyme. These ribozyme transduced cells became vincristine-sensitive concomitant with the decreases in MDR1 expression, P-gp amount and drug efflux pump function. Among the ribozymes tested, the anti-MDR1 ribozyme against the translation-initiation site exhibited the strongest efficacy. This retrovirus-mediated transfer of anti-MDR1 ribozyme may be applicable to the treatment of MDR cells as a specific means to reverse resistance. PMID- 10362144 TI - Influence of MUC18/MCAM/CD146 expression on human melanoma growth and metastasis in SCID mice. AB - The cell surface glycoprotein MUC18MCAM/CD146 was originally defined as a marker of melanoma progression and has been suspected to be directly linked to the metastatic process of this malignancy. In order to address this question, 2 MCAM negative human melanoma cell lines, SK-2 and XP44RO(Mel), were transfected with MCAM-encoding cDNA. Surface MCAM expression on SK-2 and XP44RO(Mel) transfectants was similar to that observed in naturally occurring MCAM positive human melanoma cells and transfectants demonstrated MCAM-dependent increase in homotypic adhesion in vitro. The growth behavior of 7 MCAM transfectants and their respective vector controls was evaluated in SCID mice. Tumor size at 4-5 weeks after s.c. implantation was highly variable, but did not correlate with MCAM expression. Despite massive primary tumor formation at the injection site, no spontaneous metastasis was observed with any of the investigated MCAM transfectants. The influence of MCAM expression on lung metastases formation in an experimental metastasis assay was system dependent, converting only XP44RO(Mel) transfectants into metastatic cells, although increased homotypic adhesion, leading to formation of tumor cell clusters, was observed with transfectants of both cell lines in vitro. Our findings indicate that MCAM expression of human melanoma cells has an influence on later stages of the metastatic process only, namely, extravasation and establishment of new foci of growth, but is per se not sufficient for this process. PMID- 10362145 TI - Potentiation of the malignant phenotype of the undifferentiated ARO thyroid cell line by insertion of the bcl-2 gene. AB - We have reported that bcl-2 is expressed in normal human thyroid epithelium and that its expression is down-regulated in undifferentiated thyroid tumors. Production of IL-6 was concomitantly down-regulated in these forms. Based on these observations, we analyzed whether insertion of bcl-2 would reverse the highly malignant phenotype of a thyroid cell line (ARO) derived from an undifferentiated carcinoma. This cell line fails to produce Bcl-2 and IL-6. By infection with a bcl-2 retroviral vector, ARO cells expressing bcl-2 (ARObcl-2) were obtained. Compared with parental cells, expression of bcl-2 was associated with enhancement of growth potential (DNA synthesis, in vitro proliferation rate, anchorage-independent growth in semi-solid media). Chemotaxis and invasive potential in Boyden chambers were also increased. bcl-2-expressing cells showed a reduced response to apoptotic stimuli (low-serum conditions or anti-neoplastic drugs). Large branched colonies were formed in Matrigel from ARObcl-2 cells but not from parental cells. Finally, ARObcl-2 cells showed a decreased latency of tumor appearance when injected into immunodeficient mice. Potentiation of the malignant phenotype of ARO cells by bcl-2 was not ascribed to altered expression of (i) cytokine/growth factors (IL-4, IL-6, IL-8, IL-10, IL-12, TGF-alpha, TGF beta), (ii) thyroid-specific transcripts (TG, TPO, TSH-R, PIGF, PAX-8) or (iii) genes influencing tumor aggressiveness [VEGF, HMGI (Y), HMGI-C]. Our data indicate that bcl-2 potentiates the malignant phenotype of ARO cells not only by limiting the response to apoptotic stimuli but also by enhancing proliferation and tumor aggressiveness. PMID- 10362146 TI - Rapamycin-resistant phosphorylation of the initiation factor-4E-binding protein (4E-BP1) in v-SRC-transformed hamster fibroblasts. AB - Increased phosphorylation of the translational repressor protein 4E-BP1 was found in the cell line derived from the tumor induced in Syrian hamster by Rous sarcoma virus (RSV). This was accompanied by its dissociation from the complex with initiation factor eIF4E. The ribosomal S6 protein kinase p70S6k is supposed to be regulated by the same or a closely related rapamycin-sensitive signalling pathway to that which modulates 4E-BP1. Phosphorylation and activity of p70S6k were found to be also increased in RSV-transformed H19 cells that express significantly higher amounts of the Src protein (p60src) relative to the non-transformed hamster fibroblasts NIL-2. The increased activity and phosphorylation of p70S6k were blocked by rapamycin, indicating that the rapamycin-sensitive pathway is involved in its regulation in v-src-transformed hamster fibroblasts. In agreement with this, rapamycin reduced the expression of elongation factor eEF1alpha (whose translation is regulated by a rapamycin-sensitive mechanism thought to involve p70S6k) and did not affect the production of a housekeeping protein, alpha tubulin, in these cells. Synthesis of Src protein was also inhibited in cells treated with rapamycin. However, treatment of cells with a concentration of rapamycin sufficient to completely inhibit the activity and phosphorylation of p70S6k resulted in only partial de-phosphorylation of 4E-BP1 and its re association with eIF4E in the transformed cells, indicating that additional rapamycin-insensitive mechanisms/pathways are implicated in the control of 4E-BP1 phosphorylation in RSV-transformed hamster fibroblasts. Over-expression of eIF4E favours cell proliferation and can lead to a transformed phenotype, while over expression of 4E-BP1 has the opposite effect. The altered signalling to the phosphorylation of 4E-BP1 in RSV-transformed cells, which leads to its dissociation from eIF4E and thus relief of inhibition of eIF4E function, may therefore represent an important regulatory mechanism in malignant cell growth. PMID- 10362147 TI - Identification of a novel zinc finger gene, zf5-3, as a potential mediator of neuroblastoma differentiation. AB - We established a unique parental neuroblastoma cell line, NUB-7, which mimics the bipotentiality of neuroblastoma in vivo along neuronal and Schwann cell lineages following dibutyryl cAMP and retinoic acid treatments, respectively. Differential display identified a putative novel zinc finger gene as a potential differentiation-responsive gene coincident with retinoic acid treatment of NUB-7. This cDNA clone, now designated zf5-3, was mapped to chromosome 19 using somatic cell hybrids, and a larger cDNA clone further localized this gene to band 13.1 13.2 by fluorescent in situ hybridization. zf5-3 possesses 4 characteristic zinc finger DNA-binding motifs as determined by its nucleic acid and proposed amino acid sequence. Expression of zf5-3 is restricted to fetal neuronal, hepatic and renal tissues and their tumor-derived cell lines, including 8/9 neuroblastomas and 2/2 malignant rhabdoid tumors of kidney. The restricted expression in the kidney of zf5-3 to collecting tubules and ureter epithelium is suggestive of an ectodermal histogenesis of malignant rhabdoid tumors of kidney. During development of the fetal human brain, high levels of zf5-3 mRNA are restricted to the mitotically active, undifferentiated neuroblasts. Morphological evidence of overt differentiation was generally accompanied by a marked loss in zf5-3 expression. Therefore, the neuronal tissue expression profile and the down regulation coincident with retinoic acid-induced neuroblastoma maturation implicate zf5-3 as a potential mediator of their differentiation. PMID- 10362148 TI - Cytotoxic T lymphocytes define multiple peptide isoforms derived from the melanoma-associated antigen MART-1/Melan-A. AB - Peptides derived from the melanoma-associated MART-1/Melan-A antigen are currently implemented in immunotherapy for inducing or augmenting T-cell responses directed against peptides expressed by autologous tumor cells in HLA A2+ patients with melanoma. Here, we describe the specificity of the T-cell clone SK29-FFM1.1, which secretes GM-CSF in response to a panel of synthetic MART 1/Melan-A-derived peptides, including the naturally presented ILTVILGVL(32-40), but exhibits cytotoxicity and IFN-gamma secretion exclusively to the MART-1/Melan A derived peptide AAGIGILTV(27-35). In addition, cytotoxic T-lymphocyte (CTL) clone SK29-FFM1.1 recognizes 3 different naturally processed and presented peptides on HLA-A2+ MART-1/Melan-A+ melanoma cells, as defined by cytotoxicity and IFN-gamma and GM-CSF secretion. Processing and presentation of MART-1/Melan-A peptides appears to be different in cells of non-melanocytic origin, as shown by the characterization of naturally presented peptides displayed by HLA-A2+ colorectal cancer cells transduced with a MART-1/Melan-A gene-containing retrovirus. Our data suggest that multiple epitopes, including ILTVILGVL and different isoforms of AAGIGILTV derived from MART-1/Melan-A may be naturally presented by melanoma cells to the immune system. PMID- 10362150 TI - Poststerilization regret: findings from the United States Collaborative Review of Sterilization. AB - OBJECTIVE: To evaluate the cumulative probability of regret after tubal sterilization, and to identify risk factors for regret that are identifiable before sterilization. METHODS: We used a prospective, multicenter cohort study to evaluate the cumulative probability of regret within 14 years after tubal sterilization. Participants included 11,232 women aged 18-44 years who had tubal sterilizations between 1978 and 1987. Actuarial life tables and Cox proportional hazards models were used to identify those groups at greatest risk of experiencing regret. RESULTS: The cumulative probability of expressing regret during a follow-up interview within 14 years after tubal sterilization was 20.3% for women aged 30 or younger at the time of sterilization and 5.9% for women over age 30 at sterilization (adjusted relative risk [RR] 1.9; 95% confidence interval [CI] 1.6, 2.3). For the former group, the cumulative probability of regret was similar for women sterilized during the postpartum period (after cesarean, 20.3%, 95% CI 14.5, 26.0; after vaginal delivery, 23.7%, 95% CI 17.6, 29.8) and for women sterilized within 1 year after the birth of their youngest child (22.3%, 95% CI 16.4, 28.2). For women aged 30 or younger at sterilization, the cumulative probability of regret decreased as time since the birth of the youngest child increased (2-3 years, 16.2%, 95% CI 11.4, 21.0; 4-7 years, 11.3%, 95% CI 7.8, 14.8; 8 or more years, 8.3%, 95% CI 5.1, 11.4) and was lowest among women who had no previous births (6.3%, 95% CI 3.1, 9.4). CONCLUSION: Although most women expressed no regret after tubal sterilization, women 30 years of age and younger at the time of sterilization had an increased probability of expressing regret during follow-up interviews within 14 years after the procedure. PMID- 10362149 TI - Cytotoxicity with Auger electron-emitting radionuclides delivered by antibodies. AB - We investigated the in vitro cytotoxic potential of Auger electron-emitting radionuclides delivered to the cytoplasm or, more specifically, to lysosomes, via antibodies. The antibody (Ab) used was LL1, which is specific for CD74, an epitope of the major histocompatibility complex (MHC) class II antigen invariant chain, Ii, present on the cell surface. It is taken up in large amounts, approximately 10(7) Ab molecules per cell per day, and delivered to lysosomes. The radioisotopes tested included (111)In, 99mTc and 125I. With sufficient specific activity, approximately 10 mCi/mg Ab, all of these isotopes were potent cytotoxic agents. 125I was active only if a "residualizing" form was used, meaning a form that is trapped within cells after catabolism of the Ab to which it was conjugated (conventional oxidative iodination produces a non-residualizing label). The conjugates of (111)In and 99mTc used are known to be residualizing. One hundred percent cell kill in vitro was obtained with (111)In and 125I, under conditions in which a non-reactive control Ab, conjugated in the same way, produced no significant toxicity. 99mTc was also potent and specific, but appeared somewhat less active than the other isotopes under the conditions evaluated. Although few Abs are accreted by cells at the same rate as LL1, it may be possible to use other Abs to deliver similar amounts of radioactivity, if Abs with higher specific activity can be produced. Such conjugated radioisotopes may be useful for attacking tumor cells in vivo, particularly for single cells or micrometastases. PMID- 10362151 TI - Contraceptive effectiveness of two spermicides: a randomized trial. AB - OBJECTIVE: We conducted a multinational randomized trial to determine whether a spermicidal film containing 72 mg of nonoxynol-9 per film was at least as effective in preventing pregnancy as a foaming tablet containing 100 mg of nonoxynol-9 per tablet. METHODS: Between September 1995 and July 1997, 765 women aged 18-35 years who had no evidence of subfecundity were randomly assigned to use one of the two spermicides as their only contraceptive method at every coital act for 28 weeks. Participants were asked to keep coital diaries throughout the study period. Pregnancy tests were performed on a scheduled basis. Each participant was followed for 28 weeks or until she stopped considering the spermicide as her primary method of contraception. RESULTS: The Kaplan-Meier estimate of the 6-month probability of pregnancy during typical use of the spermicide was 28.0% in the tablet group and 24.9% in the film group (P = .78, one-tailed test). The study had nearly 75% power to have detected a difference of seven percentage points between groups. Results were almost identical when the analysis included only months when the participants reported use of the spermicide during every coital act. Reported levels of sexual activity and compliance with use of the spermicide were high in both groups. CONCLUSION: The contraceptive effectiveness of these two spermicidal products appeared similar. Both products were associated with a fairly high risk of pregnancy in this young, highly sexually active population. PMID- 10362152 TI - Methotrexate compared with mercaptopurine for early induced abortion. AB - OBJECTIVE: To compare the antimetabolites methotrexate and 6-mercaptopurine as single-agent medical abortifacients using clinical and immunohistochemical analyses. METHODS: Twenty-seven women with gestations less than 7 weeks from the last menstrual period (LMP) were randomized to receive intramuscular methotrexate, 50 mg/m2, or oral 6-mercaptopurine, 200 mg. Forty-six additional women received methotrexate after randomization was discontinued. Women returned at 2-week intervals. Those without fetal cardiac activity were followed until complete abortion. Those with fetal cardiac activity were considered failures and underwent suction abortions. Tissue collected at the time of suction abortion was analyzed with the cell-proliferation immunohistochemical assay Ki-67. RESULTS: All 12 women in the 6-mercaptopurine group had fetal cardiac activity at follow up and underwent suction abortion; therefore, this arm of the study was discontinued. Six of the 61 women who received methotrexate had fetal cardiac activity at follow-up and also underwent suction abortion. Fetal cardiac activity was present after methotrexate in three of 55 women at less than 6 weeks from the LMP and in three of six between 6 and 7 weeks from the LMP (P < .01). Women who aborted after methotrexate started bleeding on day 19 (standard deviation [SD] 7.8), bled for 9 days (SD 4.0), and used minimal pain medications. Tissues exposed to methotrexate showed decreased Ki-67 activity compared with tissues exposed to 6-mercaptopurine (P = .003). CONCLUSION: In oral doses of 200 mg, 6 mercaptopurine did not induce early abortion. A single intramuscular dose of methotrexate used without prostaglandins induced abortion in most women at gestational ages of less than 6 weeks. Ki-67 activity was lower in a small sample of fetal tissues exposed to methotrexate than in tissues exposed to 6 mercaptopurine. PMID- 10362153 TI - Patients' understanding of medical risks: implications for genetic counseling. AB - OBJECTIVE: To assess patients' ability to compare magnitudes of Down syndrome risk at maternal ages of 35 and 40 years, expressed as rates or as proportions. METHODS: We used a self-administered, anonymous questionnaire that posed the same comparison in two different formats: 2.6 versus 8.9 per 1000 women (rates) and one in 384 versus one in 112 women (proportions). The study setting included several university-affiliated obstetrics and gynecology outpatient clinics in San Francisco, California. A total of 633 women, whose primary languages were English, Spanish, or Chinese, participated. The main outcome measure was correct identification of the larger of two risks. RESULTS: Women were more successful with rates (463 of 633 respondents, 73%) than with proportions (353 of 633 respondents, 56%). A paired analysis, in which each woman served as her own control, found risk assessment to be significantly better with rates than with proportions (P < .001). Women with little formal education had difficulty understanding risks framed either way. CONCLUSION: The traditional use of proportions to express risk in genetic counseling lacks scientific basis. Rates were easier to understand than proportions, regardless of respondents' age, language, and education. PMID- 10362154 TI - Quality of life among women undergoing hysterectomies. AB - OBJECTIVE: To measure the association between gynecologic conditions and quality of life in women before hysterectomy. METHODS: We retrospectively identified 482 women who had hysterectomies for nononcologic and nonemergency indications in one of nine capitated medical groups in Southern California between 1993 and 1995. Their symptoms and quality of life before hysterectomy were assessed by medical record review and telephone interview. Women were placed into four symptom-based groups (pain, bleeding, pelvic discomfort, and asymptomatic groups) and compared across six quality-of-life scales. RESULTS: Women with primary pain conditions reported the highest average role impairment compared with women with primary bleeding, pelvic discomfort, or asymptomatic conditions (8.6 days/month versus 5.0, 2.5, and 1.9 days/month, respectively; P < .05). On the five 0 to 100-point quality-of-life scales, women with primary pain conditions, compared with women with bleeding, pelvic discomfort, or asymptomatic conditions, had the highest mean levels of sexual impairment (71.5 versus 54.1, 29.6, and 17.9, respectively; P < .05) and mood impairment (55.2 versus 45.2, 34.6, and 38.1, respectively; P < .05), the poorest perception of general health (74.4 versus 60.7, 44.1, and 49.4, respectively; P < .05), and the greatest increase in severity of symptoms before hysterectomy (77.2 versus 68.7, 61.5, and 57.1, respectively; P < .05). CONCLUSION: Women's primary symptoms before hysterectomy are associated differentially with varying levels of impairment. Standardized measurement of quality of life among women with gynecologic complaints that lead to hysterectomy might help in the development of treatment guidelines and in the assessment of appropriateness and outcomes of care for those women. PMID- 10362156 TI - Predictive value for infection of febrile morbidity after vaginal surgery. AB - OBJECTIVE: To determine the screening value of febrile morbidity for detecting infections after vaginal surgery. METHODS: A cohort of 431 consecutive women had vaginal surgery at the M. S. Hershey Medical Center from September 1988 through June 1995. Outcomes of febrile morbidity and infection were analyzed. RESULTS: Fifty-four of 431 patients (12.5%) had febrile morbidity. Thirty-five infections (8.1%) were identified, of which only 13 were accompanied by febrile morbidity. Forty-one patients (9.5%) had unexplained fevers. The sensitivity of febrile morbidity for postoperative infection was 40%, specificity was 98%, positive predictive value was 26%, and negative predictive value was 94%. Stepwise logistic regression found blood loss (odds ratio 1.001/mL; confidence interval 1.0001-1.0035), uterine weight (0.987/g; 0.976-0.999), and parity (1.570; 1.146 2.050) as significant independent variables for developing fever. Patient weight (0.984/lb; 0.971-0.998) and type of procedure (2.16; 2.12-6.38) were confirmed as significant independent variables for postsurgical infections. CONCLUSION: Febrile morbidity had limited value as a screening test for postoperative infection, with poor sensitivity and positive predictive value after vaginal surgery. PMID- 10362155 TI - Delay in gynecologic surgical treatment: a comparison of patients in managed care and fee-for-service plans. AB - OBJECTIVE: To determine whether membership in a managed care organization is associated with a delay in receiving definitive surgical treatment for benign gynecologic or gynecologic oncologic diseases. METHODS: Four hundred patients who had definitive surgery between 1994 and 1997 were divided into those with benign gynecologic (n = 207) and gynecologic oncologic diagnoses (n = 193). Each group was subdivided into managed care patients and fee-for-service patients. Subgroups were analyzed for delay in surgical treatment, emergency room visits, length of stay, age, clinic visits, prior evaluation, prior treatment, second opinions, operating room time, estimated blood loss, and surgical complications. RESULTS: There were 122 managed care and 85 fee-for-service patients with benign gynecologic diagnoses. The time from initial presentation to the date of definitive surgery was significantly longer for the managed care patients (133.7 +/- 21 days compared with 84.9 +/- 12.8 days, P = .03). Of the 193 patients with gynecologic cancer 96 were in the managed care group and 97 were under fee-for service arrangements. There was no significant difference in the time from initial presentation to the date of definitive surgery between these two groups (35.7 +/- 7.4 days compared with 20.5 +/- 2.5 days, P = .29). There were no significant differences between groups in emergency room or clinic visits, prior evaluations or treatments, or surgical complications when stratified by diagnosis. The mean age of managed care patients was significantly lower than that of fee-for-service patients for gynecologic diagnoses (46.4 +/- 9.7 years compared with 56.5 +/- 14.9 years, P < .001), and gynecologic oncologic diagnoses (47.5 +/- 13.2 years compared with 60.9 +/- 15.8 years, P < .001). CONCLUSION: Membership in a managed care organization is associated with a delay in receiving definitive surgical care for benign gynecologic, but not gynecologic oncologic, diseases. PMID- 10362157 TI - The external urethral barrier for stress incontinence: a multicenter trial of safety and efficacy. Miniguard Investigators Group. AB - OBJECTIVE: To assess the efficacy and safety of an external urethral barrier for the management of mild to moderate stress urinary incontinence in adult women. METHODS: Four hundred eleven women with the symptom of stress urinary incontinence in 12 United States centers participated. Additional inclusion and exclusion criteria were applied before protocol device use, and ultimately 390 subjects began device use. Outcome measures for efficacy and safety were assessed. Efficacy was evaluated by the number of leakage episodes using a voiding diary, subjective urinary leakage severity, incontinence impact scores, and pad testing. Safety was evaluated by symptom assessment, urinalysis, urine culture, measurement of postvoid residual urine volume, vulvar cytology, vaginal culture, and (n = 81) cystometric testing. RESULTS: Efficacy was indicated by statistically significant reductions in the number of leakage episodes, subjective leakage severity scores, incontinence impact scores, and pad-test loss during device use. The data also indicated that the device was safe, as evidenced by the lack of statistically significant changes in the percentage of subjects with urinary tract infections during device use or in postvoid residual urine volume and cystometric indices. Symptoms of vulvar irritation or lower urinary tract discomfort occurred in a small percentage of subjects but were generally transient, and only three women discontinued using the device. CONCLUSION: The external urethral barrier appears to be a safe nonsurgical alternative to absorbent products for the management of mild to moderate stress urinary incontinence in adult women. PMID- 10362159 TI - Birth weight for gestational age of Mexican American infants born in the United States. AB - OBJECTIVE: To develop a reference for birth weight for gestational age to identify Mexican American infants born in the United States who are small or large for gestational age. METHODS: Reference percentiles were developed for Mexican American and non-Hispanic white births, using national vital statistics from 1992-1994 for Mexican Americans (n = 1,197,916) and 1994 for non-Hispanic whites (n = 2,238,457). Birth weights and gestation from the last menstrual period were taken from birth certificates. Smoothed curves were fit, using unweighted fourth-degree polynomial equations, for the tenth, 50th, and 90th percentiles by gender and parity. RESULTS: Mexican American infants were heavier than non-Hispanic white infants between 30 and 37 weeks' gestation for all parities and both genders. However, at term there was consistent crossover. Non Hispanic white infants were heavier at or after 37 through 42 weeks' gestation, whereas the growth of Mexican American infants appeared to slow. Beginning at 37 weeks, the differences in weights of infants of primiparas increased to more than 100 g by 40 weeks; the differences were only slightly less for infants of multiparas. CONCLUSION: Given differences in distribution of birth weights for gestational age between Mexican Americans and non-Hispanic whites, the ability to recognize fetal growth restriction (FGR) or excessive growth is questionable. These data provide a reference for Mexican Americans for clinical use and for future studies in identifying infants at risk for FGR or overgrowth. PMID- 10362158 TI - A bladder-neck support prosthesis for women with stress and mixed incontinence. AB - OBJECTIVE: To evaluate the safety and efficacy of a bladder-neck support prosthesis, a vaginal device designed to support the bladder neck, in women with genuine stress and mixed incontinence. METHODS: For enrollment, incontinent women underwent a history, physical examination including cotton-swab test, urinalysis, postvoid residual urine, and multichannel urodynamic testing. Those with genuine stress incontinence or mixed incontinence and urethral hypermobility completed 7 day bladder diaries, the Incontinence Impact Questionnaire, and underwent standardized pad tests. They were fitted with a prosthesis and seen weekly to optimize fit and efficacy. At week 5, they underwent repeat evaluations with the best-fitting prosthesis in place. RESULTS: Seventy women were enrolled and 53 completed the 1-month study (29 genuine stress incontinence, 24 mixed incontinence). The mean ages were 50.4 years for genuine stress incontinence (range 24-76) and 55.7 years for mixed incontinence (range 30-88). A statistically significant reduction in incontinence was noted on pad testing (genuine stress incontinence, mean 46.6-16.6 g; mixed incontinence, mean 31.9-6.8 g) and in the bladder diary (genuine stress incontinence, mean 28.6-7.8 losses per week; mixed incontinence, mean 30.2-15 losses per week). Quality-of-life scores improved in both groups. With the device in place, urodynamic testing indicated normalization of urethral function without evidence of outflow obstruction. Subjects found the device comfortable, easy to use, and convenient. Side effects included five urinary tract infections and 23 cases of vaginal mucosal soreness or mild irritation. CONCLUSION: The bladder-neck support prosthesis significantly reduced involuntary urine loss in women with stress and mixed incontinence. PMID- 10362160 TI - Cardiac compliance in fetuses of diabetic women. AB - OBJECTIVE: To examine possible changes in cardiac function in fetuses of pregestational diabetic mothers. METHODS: We conducted a prospective longitudinal study of 31 women whose pregnancies were between 22 weeks' gestation and term, and who had pregestational diabetes. All diabetic women included in the study had glycosylated hemoglobin lower than 6.5%. All patients included in the study had an early ultrasound confirming gestational age. Doppler studies of the blood flow through the mitral and tricuspid valves were done every 4 weeks using a pulsed wave Doppler ultrasound device with a 3.5- or 5-MHz transducer. The following indices were calculated from the flow velocity waveforms: the peak velocity during the rapid ventricular filling (E wave) and during the atrial systole (A wave), and the ratio between these velocities (E/A ratio); and the velocity time integral of the atrioventricular blood flow (this integral correlates with volume flow). A comparison between the Doppler indices obtained in fetuses of diabetic women and of normal women was made by using the Mann-Whitney test. RESULTS: Each patient had four to five fetal echocardiographic examinations at 22, 26, 30, 34, and 38 weeks' gestation. The E/A ratio of the mitral and tricuspid valves did not increase in fetuses of diabetic women during the third trimester and was significantly higher in fetuses of nondiabetic women compared with fetuses of diabetic women at 34 and 38 weeks' gestation. The velocity time integral of the mitral and tricuspid valves multiplied by heart rate was higher, but not significantly, in fetuses of nondiabetic women compared with fetuses of diabetic women at 34 and 38 weeks' gestation. The E-wave of the mitral and tricuspid valves increased in both groups throughout gestation. The A-wave of the mitral and tricuspid valves increased only in fetuses of diabetic women throughout the third trimester and was significantly higher at 34 and 38 weeks' gestation compared with fetuses of nondiabetic women. CONCLUSION: Differences in atrioventricular blood flow patterns between fetuses of diabetic women and normal fetuses do not necessarily result from differences in cardiac compliance. PMID- 10362161 TI - Neonatal lupus erythematosus: results of maternal corticosteroid therapy. AB - OBJECTIVE: To assess the possibility of preventing cardiac or cutaneous manifestations of neonatal lupus erythematosus or treating the fetus with congenital heart block by administering corticosteroid therapy to the mother. METHODS: Eighty-seven offspring of 40 anti-Ro/SSA-positive mothers, followed up from 1979 to 1996, were evaluated. Autoantibodies against Ro/SSA and La/SSB antigens were detected by immunodiffusion and enzyme-linked immunosorbent assay. RESULTS: None of 26 neonates whose mothers received corticosteroid maintenance therapy initiated before 16 weeks' gestation demonstrated congenital heart block, whereas 15 of 61 neonates whose mothers received no corticosteroids during pregnancy or began receiving steroid therapy after 16 weeks' gestation had congenital heart block. Complete congenital heart block, once developed, did not respond to corticosteroid treatment in utero. Four infants whose mothers received steroid treatment before 16 weeks' gestation had skin lesions of neonatal lupus erythematosus. CONCLUSION: Once established, complete congenital heart block was irreversible and maternal corticosteroid therapy did not effectively prevent cutaneous lupus erythematosus. However, prenatal maintenance therapy with prednisolone or betamethasone given to the mother starting early in pregnancy (before 16 weeks' gestation) might reduce the risk of developing antibody mediated congenital heart block in the offspring. PMID- 10362162 TI - Nitric oxide in the uteroplacental, fetoplacental, and peripheral circulations in preeclampsia. AB - OBJECTIVE: Altered production of nitric oxide by the vascular endothelium may influence the pathogenesis of preeclampsia. The aim of this study was to measure circulating levels of nitric oxide metabolites (nitrites) in the uteroplacental, fetoplacental, and peripheral circulation of preeclamptic pregnancies compared with normotensive controls. METHODS: Fifteen women with preeclampsia were compared with 16 women with normotensive pregnancies. At cesarean, blood samples were taken from the uterine vein draining the placental site, the umbilical vein, and the antecubital vein after delivery of the baby but before delivery of the placenta. Plasma nitrites were measured using the Greiss reaction after conversion of plasma nitrates to nitrites using nitrate reductase. RESULTS: Nitric oxide metabolites were higher in the uteroplacental (P < .01), fetoplacental (P < .001), and peripheral (P < .02) circulations in samples from preeclamptic pregnancies compared with control pregnancies. In samples from the fetoplacental circulation only, nitric oxide metabolite levels were negatively correlated with gestational age (r = -.489, P < .01) and birth weight (r = -.544, P < .004). Nitric oxide metabolite levels were not significantly correlated with blood pressure, placental weight, or maternal age. CONCLUSION: In established preeclampsia, production of nitric oxide was higher in the uteroplacental, fetoplacental, and peripheral circulation than in normotensive pregnancies. This increase may be part of a compensatory mechanism to offset the pathologic effects of preeclampsia. PMID- 10362163 TI - Amniotic fluid concentrations of adrenomedullin in preterm labor. AB - OBJECTIVE: To determine whether adrenomedullin levels in amniotic fluid were associated with preterm labor. METHODS: We measured immunoreactive adrenomedullin in amniotic fluid collected by amniocentesis from 36 women with clinical diagnosis of preterm labor or preterm premature rupture of membranes (PROM) and from 18 normal pregnant women. RESULTS: Amniotic fluid from cases of PROM and failure to respond to tocolysis were associated significantly with higher amniotic fluid adrenomedullin concentrations (177.0 +/- 22.5 pg/mL and 182.7 +/- 22.0 pg/mL, respectively, P < .01) than that from uncomplicated pregnancies (101.2 +/- 28.1 pg/mL) or preterm labor responsive to tocolysis (102.3 +/- 26.8 pg/mL). CONCLUSION: Amniotic fluid adrenomedullin is higher than normal in cases of PROM and preterm labor unresponsive to tocolysis, perhaps indicating enhanced synthesis from placenta or fetal membranes being stimulated by bacterial products. PMID- 10362164 TI - Clearance of fetal products and subsequent immunoreactivity of blood salvaged at cesarean delivery. AB - OBJECTIVE: To determine if fetal products can be detected after postplacental, intraoperative blood salvage, and if the product is immunoreactive with maternal serum. METHODS: We suctioned the shed blood of 27 term gravidas with intact membranes who had cesareans, beginning 4 minutes after placenta removal, into a COBE BRAT-2 salvage system (COBE Cardiovascular, Arvada, CO). Preoperative maternal and fetal cord blood samples were collected. Preprocessing and postprocessing salvaged blood was analyzed for alpha-fetoprotein (AFP), hemoglobin, hematocrit, and plasma-free hemoglobin. Papanicolaou smears and immunodiffusion using Ouchterlony methods for detection of protein-protein interactions were run on maternal serum. Postprocess salvaged blood was subjected to Kleihauer-Bethke tests, typed, and crossmatched with maternal serum, including mixed fields. No women were transfused. RESULTS: Ten of 27 women shed enough postprocess salvaged blood for analysis. Alpha-fetoprotein was cleared, but Kleihauer-Bethke analyses were positive in all postprocessing specimens. Anucleate squamous cells were detected by Papanicolaou smears in four of ten preprocessed specimens, with one cleared by processing. No antigen-antibody reaction between maternal and preprocessed or postprocessed salvaged blood was found by the Ouchterlony method. Crossmatching of the final product with maternal serum was successful, with negative mixed fields in all cases. CONCLUSION: Fetal debris was present in blood salvaged 4 minutes after removal of placenta. Despite clearance of humoral material, fetal blood cells were detectable in all postprocess salvaged blood. The product was compatible with maternal blood by crossmatching and its supernate did not immunoreact with maternal serum. PMID- 10362165 TI - Amniotic fluid embolism: decreased mortality in a population-based study. AB - OBJECTIVE: To examine the risk factors and pregnancy outcomes associated with 53 cases of amniotic fluid embolism that occurred in California during the 2-year period January 1, 1994 to December 31, 1995. METHODS: Data were obtained from a computerized database that contains linked records from the vital statistics birth certificate and hospital discharge summaries of both mother and newborn. This database covered all singleton deliveries that occurred in 328 civilian acute-care hospitals in California, which represented 98% of all deliveries in California. All cases of amniotic fluid embolism were examined for other pregnancy complications. RESULTS: There were 1,094,248 deliveries during that 2 year period. Fifty-three singleton gestations had the diagnosis of amniotic fluid embolism, for a population frequency of one per 20,646 deliveries. Fourteen women with amniotic fluid embolism died, for a maternal mortality rate of 26.4%. There were 35 (66%) diagnoses of disseminated intravascular coagulation (DIC), 38 (72%) diagnoses of hemorrhage, and 25 (47%) diagnoses of obstetric shock. Among the 14 women who died, the frequency of DIC (79%) and hemorrhage (71%) was not different compared with that of the survivors (62% and 72%, respectively), but obstetric shock was higher (86%, P = .02) than in survivors (33%). The average maternal length of stay for survivors was 6.5 days (range 3-27 days, median 5 days). The cesarean rate was 60% and the frequency of fetal distress was 49%. CONCLUSION: In this population-based study of reported cases of amniotic fluid embolism, the maternal mortality rate (26.4%) was significantly less than previously reported and might reflect a more accurate population frequency. In addition, patients who survived and patients who died had similar pregnancy complications, suggesting that amniotic fluid embolism was present in all cases and not limited to those who died. PMID- 10362166 TI - Dietary therapy for gestational diabetes: how long is long enough? AB - OBJECTIVE: To determine the length of time required for dietary therapy alone to effect good glycemic control and whether the need for insulin treatment can be predicted at diagnosis of gestational diabetes mellitus (GDM). METHODS: Women with GDM were treated with dietary therapy for 4 weeks. Each measured her blood glucose using a memory-based reflectance glucometer, and those in poor glycemic control (mean glucose exceeding 105 mg/dL) after 4 weeks of dietary therapy were prescribed insulin. Women were stratified by fasting plasma glucose value of 3 hour glucose tolerance tests (GTTs). RESULTS: Women with fasting glucose at or below 95 mg/dL were significantly more likely to achieve good glycemic control after 2 weeks of dietary therapy than were those with values above 95 mg/dL whose control did not improve during the study. Receiver operating characteristic (ROC) analysis determined that fasting values of GTT between 91 and 95 mg/dL best predicted that insulin would be needed for good glycemic control. CONCLUSION: Women with GDM should be prescribed dietary therapy alone for at least 2 weeks before they are prescribed insulin. In those with fasting glucose above 95 mg/dL, insulin may be prescribed after 1 week of dietary therapy, or at diagnosis. PMID- 10362167 TI - Hepatitis B vaccination in pregnancy: factors influencing efficacy. AB - OBJECTIVE: To determine seroprotective antibody response after hepatitis B vaccination during pregnancy and to assess factors influencing the rate of maternal seroprotection. METHODS: Records of 80 healthy gravidas who elected hepatitis B vaccination during pregnancy, after being identified as hepatitis B surface antigen (HbsAg) and antibody (HbsAb) negative on initial prenatal screen, were analyzed retrospectively. Each gravida was begun on a series of three recombinant hepatitis B vaccines at 0, 1, and 6 months. At 36-40 weeks' gestation, all gravidas were rescreened for seroprotective levels of HbsAb using qualitative enzyme-linked immunosorbent assay analysis. The women were grouped by maternal age (less than 25 years or at least 25 years), smoking history, maternal weight, body mass index (BMI) (less than 30, at least 30, less than 34, or at least 34), number of vaccinations received, race-ethnicity, gestational age at vaccination, and vaccination-to-rescreening interval. Data were compared by t test, chi2 test, or Fisher exact test. Stepwise logistic regression analysis was done. RESULTS: At rescreening, 39 (49%) of the 80 women had seroprotective HbsAb conversion. After two vaccinations, obese women (BMI at least 30) (P = .04), women at least 25 years old (P = .04), and women with smoking histories (P = .005) were significantly less likely to respond to the vaccine. Logistic regression analysis for predicting failure of seroprotective response after two vaccinations showed significantly increased odds for severe obesity with BMI at least 34 (odds ratio [OR] 16.2; 95% confidence interval [CI] 1.7, 154.7), smoking history (OR 7.5; 95% CI 2.0, 27.7), and age at least 25 years (OR 3.9; 95% CI 1.1, 14.4). CONCLUSION: Maternal obesity, advancing age, and smoking have negative influences on the efficacy of hepatitis B vaccination in pregnant women. PMID- 10362169 TI - Epidural analgesia and active management of labor: effects on length of labor and mode of delivery. AB - OBJECTIVE: To determine whether cervical dilatation at the time of placement of patient-requested epidural affects cesarean rates or lengths of labors in actively managed parturients. METHODS: The charts of 255 women randomized to active management of labor (n = 125) or control protocols (n = 130) were reviewed and stratified to early epidural placement (up to 4 cm cervical dilatation) versus late placement (more than 4 cm). RESULTS: Women with early epidural placement had shorter labors than those with late placement (11.6 +/- 4.6 versus 13.2 +/- 5.6 hours; P = .02). Active management reduced the length of labor compared with controls regardless of epidural timing, with a reduction of 1.4 hours in early epidural placement (10.9 +/- 4.7 versus 12.3 +/- 4.3 hours; P = .04) and 3.6 hours in those with later placement (11.0 +/- 3.6 versus 14.6 +/- 6.2 hours; P = .004). Cesarean rates did not vary significantly (early 14.5% versus late 7.9%; P = .21). Early epidural placement did not lengthen the second stage of labor or increase operative vaginal delivery rates. CONCLUSION: Early epidural placement did not affect lengths of labor or cesarean rates and was actually associated with shorter labor compared with late epidural placement. Women managed actively in labor, regardless of timing of epidural placement, had shorter labors than controls. PMID- 10362168 TI - Nitric oxide-mediated effects on myometrial contractility at term during prelabor and labor. AB - OBJECTIVE: To assess the existence of a nitric oxide (NO) system in the human myometrium and the effects of mediators of this system on contractile activity in vitro. METHODS: Myometrial tissue was obtained before the onset of labor and during labor at term. Production of NO was assessed by the use of nicotinamide dinucleotide phosphate diaphorase staining and by immunoblots for NO. Effects of NO were examined by adding L-arginine (the substrate for NO synthesis); N(G) nitro-L-arginine methyl ester (an inhibitor of NO synthase); two NO donors, sodium nitroprusside and spermine NONOate; as well as 8-bromo cyclic guanosine monophosphate (8-bromo cGMP) (a second messenger analogue) to organ baths. RESULTS: Myometrial NO production was indicated by positive nicotinamide adenine dinucleotide phosphate diaphorase staining. Immunoblots detected endothelial NO synthase, whereas only a weak signal for inducible NO synthase was seen. The addition of L-arginine (10(-4)-10(-3) mol/L) did not result in any change of contractility. N(G)-nitro-L-arginine methyl ester (10(-3) mol/L) caused a minor increase of contractility in half of the specimens. Sodium nitroprusside, spermine NONOate, and 8-bromo cGMP resulted in a concentration-dependent inhibition of contractility (10(-7)-10(-6) mol/L for sodium nitroprusside, 10(-6) 10(-5) mol/L for spermine NONOate, and 10(-5)-10(-3) mol/L for 8-bromo cGMP). However, at 10(-5)-10(-4) mol/L, sodium nitroprusside exhibited a dose-dependent increase in the frequency of contractions. Women in prelabor did not differ from those in active labor. CONCLUSION: The myometrium produces NO at term. Nitric oxide inhibits myometrial contractile activity. The responsiveness to NO is similar in nonlaboring and laboring women. PMID- 10362170 TI - Racial differences in hormone replacement therapy prescriptions. AB - OBJECTIVE: To examine racial differences in hormone replacement therapy (HRT) use by analyzing the relative risks and rates of HRT prescriptions for black and white women. METHODS: Data on visits to hospital outpatient departments and office-based physicians by black and white women aged 45-64 years were obtained from 25,203 visits sampled in the 1993-1995 National Ambulatory Medical Care Surveys and National Hospital Ambulatory Medical Care Surveys. The relative effect of race on the provision of an HRT prescription at an ambulatory visit was estimated by controlling confounders using logistic regression. Population-based rates of physician visits and visits with HRT prescriptions were also calculated to address issues involving access to care. RESULTS: Approximately 98,787,000 annual visits were made by black and white women 45-64 years of age, 9.2% of which involved prescriptions for HRT. The percentage of visits by black women in which prescriptions for HRT were reported (4.5%) was roughly half that of white women (9.7%). The association persisted after controlling for type of physician, practice type, geographic region, payment source, and non-HRT prescription(s) (odds ratio 2.1; 95% confidence interval 1.5, 2.9). The rate of ambulatory care among black women (3.82 visits per year per woman) was virtually identical to that of white women (3.94 visits per year), whereas the rate of visits with HRT prescriptions for white women was twice as high as for black women (0.38 and 0.17 visits per year, respectively). CONCLUSION: Apparent racial differences in HRT use persist after controlling for physician and visit factors not explored in previous studies. PMID- 10362171 TI - Endometrial thickness in tamoxifen-treated patients: an independent predictor of endometrial disease. AB - OBJECTIVE: To assess the independent contribution of transvaginal ultrasound in identifying women at risk for endometrial disorders, and determine whether a cutoff value identifies women who need endometrial histologic assessment. METHODS: Postmenopausal women with breast cancer who were receiving tamoxifen, with ultrasonographic endometrial thickness greater than 4 mm or vaginal bleeding, had hysteroscopy with selective endometrial biopsies. Endometrial thickness, duration of tamoxifen therapy, and endometrial histology were studied. Parametric and nonparametric tests and logistic regression and receiver operating characteristic curves were used for statistical analysis. RESULTS: The study population consisted of 163 women, 46 with vaginal bleeding. The proportion of women with abnormal histologic findings was greater among those with endometrial thicknesses exceeding 9 mm compared with those with endometrial thicknesses 9 mm or less (60% versus 6.1%, P < .001) and among women who received tamoxifen for more than 27 months than those who received it for less time (46% versus 16%, P < .005). Logistic regression showed that endometrial thickness greater than 9 mm and vaginal bleeding were independent predictors of abnormal findings at hysteroscopy. CONCLUSION: In women taking tamoxifen, sonographic endometrial thickness exceeding 9 mm and the presence of vaginal bleeding are independent predictors of endometrial disease. If either exists, hysteroscopy and biopsy should be done. PMID- 10362172 TI - Bone density effects of continuous estrone sulfate and varying doses of medroxyprogesterone acetate. Ogen/Provera Study Group. AB - OBJECTIVE: To establish the optimum oral daily dose of medroxyprogesterone acetate with estrone sulfate for 2 years to maintain bone density. METHODS: A multicenter, double-blind study involved 568 postmenopausal women given estrone sulfate, 1.25 mg, and randomized to receive 2.5, 5, or 10 mg of medroxyprogesterone acetate. Bone density analyses of the lumbar spine and femoral neck were done at baseline and 12 and 24 months. RESULTS: There was a significant increase from baseline to 24 months in mean lumbar spine (4.0% +/- 0.27%) and femoral neck (3.2% +/- 0.28%) bone density, with no significant differences between the treatment groups. Factors most influencing bone density changes were baseline bone density and treatment duration. Significant increases were seen in the spine over 2 years; in the hip, those occurred in the first 12 months only. In both sites, lower baseline bone density resulted in greater increases. In the spine only, no previous hormone replacement therapy, higher body mass index, more than 2 years postmenopause, and nonsmoking resulted in greater gains. Once those covariates and center-to-center variations were corrected for, in the spine, the 10-mg group had smaller increases than the other groups. Changes were unrelated to age, parity, calcium, and alcohol intakes in either site. CONCLUSION: Daily estrone sulfate, 1.25 mg, with 2.5, 5, or 10 mg medroxyprogesterone acetate was effective for preventing bone loss in postmenopausal women. PMID- 10362173 TI - Logistic regression models in obstetrics and gynecology literature. AB - OBJECTIVE: To evaluate the reporting of multivariable logistic regression analyses and assess variations in quality over time in the obstetrics and gynecology literature. METHODS: Methodologic criteria for reporting logistic regression analyses were developed to identify problems affecting accuracy, precision, and interpretation of this approach to multivariable statistical analysis. These criteria were applied to 193 articles that reported multivariable logistic regression in the issues of four generic obstetrics and gynecology journals in 1985, 1990, and 1995. Rates of compliance with the methodologic criteria and their time trends were analyzed. RESULTS: The proportion of articles using logistic regression analysis increased over time: 1.7% in 1985, 2.8% in 1990, and 6.5% in 1995 (P < .001 for trend). Violations and omissions of methodologic criteria for reporting logistic models were common. The research question, in terms of dependent and independent variables, was not clearly reported in 32.1%. The process of variable selection was inadequately described in 51.8% of the articles. Among articles with ranked independent variables, 85.1% did not report assessment of conformity to linear gradient. Tests for goodness of fit were not given in 93.2% of articles. The contribution of the independent variables could not be evaluated in 36.2% of the articles because of a lack of coding of the variables. Interactions between variables were not assessed in 86.4% of articles. Analysis of variations in the quality of logistic regression analyses over time showed no increase in reporting of the criteria concerning variable selection and goodness of fit. However, the proportion of articles reporting one quality criterion concerning interpretation of the substantive significance of independent variables showed a trend toward improvement: 42.3% in 1985, 73.6% in 1990, and 75.4% in 1995 (P = .004 for trend). CONCLUSION: The reporting of multivariable logistic regression models in the obstetrics and gynecology literature is poor, and the time trends of improvement in quality of reporting are not particularly encouraging. PMID- 10362174 TI - Risk of repetition of a severe perineal laceration. AB - OBJECTIVE: To compare the outcome of subsequent delivery in women with a history of a third- or fourth-degree laceration with outcomes in women without such a history. METHODS: This retrospective study used a perinatal database and chart review from 1978 to 1995. Only women whose first delivery was at our institution at more than 36 weeks' gestation, vaginal singleton, vertex presentation, and birth weight greater than 2500 g, with a subsequent delivery were included. The women were grouped by presence or absence of a third- or fourth-degree (severe) perineal laceration in their first delivery. The subsequent delivery was analyzed for maternal age, weight, birth weight, gestational age, method of delivery, use of episiotomy, and occurrence of a severe laceration. Comparison of data was by Fisher exact and t tests. RESULTS: Four thousand fifteen women met our starting criteria. In their first delivery, the average birth weight, use of instrumentation, and episiotomy rate were significantly higher in those women sustaining a severe laceration. When compared with women without a history of severe perineal laceration, women with such a history were at more than twice the risk for another in their subsequent delivery. The women at highest risk (21.4%) were those sustaining a laceration in their first delivery who underwent instrumental vaginal delivery with episiotomy in their subsequent delivery. When episiotomy or instrumental delivery was performed in the second vaginal birth, 52 (11.6%) of 449 women with a history of a severe perineal laceration sustained another, compared with 98 (6.5%) of 1509 without such a history (P < .001, odds ratio 1.9, 95% confidence interval 1.3, 2.7). CONCLUSION: Women delivering their second baby, and in whom episiotomy or instrumentation is used, are at increased risk of severe perineal laceration compared with women delivery spontaneously. PMID- 10362175 TI - Risk adjustment for interhospital comparison of primary cesarean rates. AB - OBJECTIVE: To create a method of controlling for case mix so that inferences could be made about variation in cesarean rates among hospitals. METHODS: A total of 160,753 births from 1991 Illinois birth certificate data were analyzed. A multivariate model of characteristics independently associated with cesarean delivery was developed from a random 25% sample, validated on the other 75%, and used to create a probability of cesarean delivery for each woman. The validated model was used to calculate a predicted primary cesarean delivery rate for the 154 hospitals in Illinois that did at least 100 deliveries per year. RESULTS: The final model included both medical and sociodemographic risk factors and predicted primary cesarean rates accurately over a full range of rates. Thirty-five hospitals (23%) had actual rates that were higher than their individual predicted 95% confidence interval (CI). Eighty-nine hospitals (58%) had actual rates within predicted CIs. Thirty hospitals (20%) had actual rates that were lower than the predicted 95% CI. Twenty-three percent of hospitals with actual rates greater than predicted rates were not in the top quartile of actual rates. Twenty-seven percent of hospitals with actual rates in the top quartile were doing cesarean deliveries appropriate for the risk status of the population served. CONCLUSION: Risk adjusting for hospital case mix more accurately identifies outlier hospitals than raw, unadjusted primary cesarean delivery rates. We believe that risk adjusting should be the first step in understanding variations in primary cesarean delivery rates. PMID- 10362176 TI - Predictors of cesarean delivery after prelabor rupture of membranes at term. AB - OBJECTIVE: To identify the significant predictors of cesarean delivery after prelabor rupture of membranes (PROM) at term. METHODS: In a multicenter study involving 72 institutions in six countries, 5041 women were randomized to induction of labor with oxytocin or prostaglandins or to expectant management. We did univariate and multivariate logistic regression analyses to determine the statistically significant independent predictors of cesarean delivery (P < .05). RESULTS: The following variables were found to be significantly associated with cesarean delivery: delivery in Israel, versus Canada (odds ratio [OR] 0.34); delivery in Australia, versus Canada (OR 1.93); nulliparity (OR 2.81); labor lasting more than 12 hours, versus less than 6 hours (OR 2.78); labor lasting 6 12 hours, versus less than 6 hours (OR 1.66); previous cesarean delivery (OR 2.75); epidural anesthesia (OR 2.66); clinical chorioamnionitis (OR 2.42); internal fetal heart rate monitoring (OR 2.19); birth weight of at least 4000 g (OR 2.07); use of oxytocin (OR 1.97); maternal age of at least 35 years (OR 1.44); latent period of at least 12 hours (OR 1.41); and meconium staining (OR 1.41). CONCLUSION: Strong predictors of cesarean delivery after PROM at term were country of birth, nulliparity, long labor, previous cesarean delivery, and epidural anesthesia. PMID- 10362177 TI - Fetal pulse oximetry: duration of desaturation and intrapartum outcome. AB - OBJECTIVE: To analyze labor outcomes in relation to masked fetal arterial oxyhemoglobin saturation values above or below 30%. METHODS: Consenting gravidas with uncomplicated pregnancies at or beyond 36 weeks' gestation underwent continuous fetal pulse oximetry. Pregnancy outcomes were compared between two groups: women with fetuses with at least one epoch of arterial oxyhemoglobin saturation below 30% (10 seconds or longer) and women with fetuses without such an episode. We also attempted to ascertain whether duration of saturation below 30% correlated with fetal compromise. RESULTS: We measured arterial oxyhemoglobin saturation in 129 fetuses, 69 (53%) of whom had at least one epoch of saturation below 30%. There were no statistically significant differences in labor and delivery outcomes between the high-saturation and low-saturation groups (eg, cesarean delivery: 13 versus 9%, P = .41; umbilical artery [UA] pH less than 7.20: 10 versus 9%, P > .999). However, as duration of fetal arterial oxyhemoglobin saturation below 30% increased from 10 seconds to longer than 9 consecutive minutes, the incidence of fetal compromise (considered present when at least one of the following criteria was met: cesarean delivery for nonreassuring fetal heart rate pattern, UA pH less than 7.20, admission to the special care nursery, or 5-minute Apgar score not more than 3) increased significantly (P = .002). The threshold duration of fetal arterial oxyhemoglobin saturation below 30% associated with increased fetal compromise was 2 minutes. CONCLUSION: Transient fetal arterial oxyhemoglobin saturation values below 30% are common during normal labor and did not predict fetal compromise. Fetal arterial oxyhemoglobin saturation values less than 30% for 2 minutes or longer might be associated with fetal compromise. PMID- 10362178 TI - Fluoroscopically guided hysteroscopic division of adhesions in severe Asherman syndrome. AB - BACKGROUND: Severe Asherman syndrome that is stage III disease according to the American Fertility Society, with obliteration of the uterine cavity and the inability to visualize isolated pockets of the intrauterine cavity, makes safe and effective hysteroscopic division of adhesions difficult, if not impossible. TECHNIQUE: A 16-gauge, 80-mm Tuohy needle is introduced into the endocervical canal alongside a 5-mm diagnostic hysteroscope. The surgeon probes the area beyond the adhesion with the needle. Ultravist 76.9% is injected through the needle under fluoroscopic and hysteroscopic control. Hidden pockets of endometrium can be located radiographically, a passageway is created using the needle, and subsequent division of adhesions is performed under direct vision with hysteroscopic scissors. EXPERIENCE: Since 1984, approximately 55 women with severe Asherman syndrome have undergone this procedure. All patients required at least two procedures, and one woman required six. There have been two cases of uneventful perforation with the Tuohy needle, and all women resumed menstruation. No serious complications have occurred. CONCLUSION: This technique provides an intraoperative fluoroscopic view of pockets of endometrium behind an otherwise blind-ending endocervical canal in women with severe Asherman syndrome, allowing guided division of adhesions and reducing the likelihood of perforation and formation of false passageways. PMID- 10362179 TI - A breast clinic in a department of obstetrics and gynecology. AB - In 1988, the Department of Obstetrics and Gynecology of the University of Southern California School of Medicine; created its own Breast Diagnostic Center for training resident physicians and providing breast care for outpatients and inpatients of Women's and Children's Hospital, Los Angeles, California. The structure and function of the Breast Diagnostic Center allow residents to be directly involved in and responsible for evaluation and care of benign breast problems and allow comprehensive breast-care education and integration of referral breast services for residents and patients. Direct faculty supervision, uniform history and physical records on printed forms, fine-needle aspirations and breast biopsies, and staff assistance with follow-up and patient tracking maximize resident physician education and experience. This departmental approach to resident physician training in breast care can be adapted to the resources and logistics of any department of obstetrics and gynecology. PMID- 10362180 TI - 50th anniversary historical article. Thrombolytic therapy in acute myocardial infarction. PMID- 10362181 TI - The multicenter study of enhanced external counterpulsation (MUST-EECP): effect of EECP on exercise-induced myocardial ischemia and anginal episodes. AB - OBJECTIVES: The purpose of this study was to assess safety and efficacy of enhanced external counterpulsation (EECP). BACKGROUND: Case series have shown that EECP can improve exercise tolerance, symptoms and myocardial perfusion in stable angina pectoris. METHODS: A multicenter, prospective, randomized, blinded, controlled trial was conducted in seven university hospitals in 139 outpatients with angina, documented coronary artery disease (CAD) and positive exercise treadmill test. Patients were given 35 h of active counterpulsation (active CP) or inactive counterpulsation (inactive CP) over a four- to seven-week period. Outcome measures were exercise duration and time to > or =1-mm ST-segment depression, average daily anginal attack count and nitroglycerin usage. RESULTS: Exercise duration increased in both groups, but the between-group difference was not significant (p > 0.3). Time to > or =1-mm ST-segment depression increased significantly from baseline in active CP compared with inactive CP (p = 0.01). More active-CP patients saw a decrease and fewer experienced an increase in angina episodes as compared with inactive-CP patients (p < 0.05). Nitroglycerin usage decreased in active CP but did not change in the inactive-CP group. The between-group difference was not significant (p > 0.7). CONCLUSIONS: Enhanced external counterpulsation reduces angina and extends time to exercise-induced ischemia in patients with symptomatic CAD. Treatment was relatively well tolerated and free of limiting side effects in most patients. PMID- 10362182 TI - EECP-enhanced external counterpulsation. PMID- 10362183 TI - Immediate exercise testing of low risk patients with known coronary artery disease presenting to the emergency department with chest pain. AB - OBJECTIVES: The purpose of this study was to demonstrate the safety and utility of immediate exercise treadmill testing (IETT) of low risk patients presenting to the emergency department with known coronary artery disease (CAD). BACKGROUND: More than 70% of the two million patients admitted to U.S. hospitals annually for suspected acute myocardial infarction (AMI) are found not to have had a cardiac event. We have previously demonstrated the safety and efficacy of IETT of selected low risk patients without known CAD presenting to the emergency department with chest pain. This study extends this approach to selected patients with a history of CAD. METHODS: One hundred patients evaluated by the chest pain emergency room to rule out AMI underwent IETT using a modified Bruce protocol upon admission to the hospital (median time <1 h). RESULTS: Twenty-three patients (23%) had positive exercise electrocardiograms (ExECGs); an uncomplicated non-Q wave AMI was diagnosed in two patients. Thirty-eight patients (38%) had negative ExECGs and 39 patients (39%) had nondiagnostic ExECGs. Of these 100 patients, 64 were discharged immediately after IETT, 19 were discharged in less than 24 h after negative serial cardiac enzymes and stable electrocardiograms and 17 were discharged after further evaluation and treatment. There were no complications from exercise testing and no late deaths or AMI during six-month follow-up. CONCLUSIONS: Immediate exercise treadmill testing of low risk patients with chest pain and known CAD is effective in further stratifying this group into patients who can be safely discharged and those who require hospital admission. PMID- 10362184 TI - Myocardial viability on echocardiography predicts long-term survival after revascularization in patients with ischemic congestive heart failure. AB - OBJECTIVES: This study was conducted to evaluate the effect of revascularization on survival in patients with congestive heart failure (CHF) due to ischemic left ventricular (LV) systolic dysfunction based on the presence of myocardial viability (MV). BACKGROUND: There are insufficient data regarding the survival benefit of revascularization in patients with CHF due to ischemic LV systolic dysfunction. METHODS: Follow-up was obtained in 87 consecutive patients with CHF due to ischemic LV systolic dysfunction (New York Heart Association [NYHA] class II-IV; LV ejection fraction <0.35) who underwent low-dose dobutamine echocardiography (DE). MV within each of 12 myocardial segments representing the LV was defined as having either: 1) normal function or mild dyssynergy at rest; 2) severe resting dyssynergy that improved on DE, or 3) worsening of function on DE except in the case of akinesia. RESULTS: At a mean follow-up of 40+/-17 months, 37 patients had received revascularization on the basis of clinical grounds, and there were 22 (25%) cardiac-related deaths. Multivariate Cox regression analysis revealed that when patients with at least five segments showing MV underwent revascularization, mortality was reduced by an average of 93% (confidence interval of 22% to 99%), which was associated with improvement in NYHA class as well as LV ejection fraction. Patients with less than five segments showing MV who underwent revascularization (and thus, showing mostly scar), and those with at least 5 segments demonstrating MV who were treated medically, had a much higher mortality. CONCLUSIONS: Revascularization produces a clear survival benefit in patients with CHF due to ischemic LV systolic dysfunction who have a significant region of the LV demonstrating MV. These data may have wide-ranging implications in the management of patients with coronary artery disease whose main clinical presentation is CHF. PMID- 10362185 TI - Low hot pain threshold predicts shorter time to exercise-induced angina: results from the psychophysiological investigations of myocardial ischemia (PIMI) study. AB - OBJECTIVES: The purpose of this study was to test whether cutaneous thermal pain thresholds are related to anginal pain perception. BACKGROUND: Few ischemic episodes are associated with angina; symptoms have been related to pain perception thresholds. METHODS: A total of 196 patients with documented coronary artery disease underwent bicycle exercise testing and thermal pain testing. The Marstock test of cutaneous sensory perception was administered at baseline after 30 min of rest on two days and after exercise and mental stress. Resting hot pain thresholds (HPTs) were averaged for the two baseline visits and divided into two groups: 1) average HPT <41 degrees C, and 2) average HPT > or =41 degrees C, to be clearly indicative of abnormal hypersensitivity to noxious heat. RESULTS: Patients with HPT <41 degrees C had significantly shorter time to angina onset on exercise testing than patients with HPT > or =41 degrees C (p < 0.04, log-rank test). Heart rates, systolic blood pressure and rate-pressure product at peak exercise were not different for the two groups. Resting plasma beta-endorphin levels were significantly higher in the HPT <41 degrees C group (5.9+/-3.7 pmol/liter vs. 4.7+/-2.8 pmol/liter, p = 0.02). Using a Cox proportional hazards model, patients with HPT <41 degrees C had an increased risk of angina (p = 0.03, rate ratio = 2.0). These differences persisted after adjustment for age, gender, depression, anxiety and history of diabetes or hypertension (p < 0.01). CONCLUSIONS: Occurrence of angina and timing of angina onset on an exercise test are related to overall hot pain sensory perception. The mechanism of this relationship requires further study. PMID- 10362186 TI - Effects of naloxone on myocardial ischemic preconditioning in humans. AB - OBJECTIVES: We attempted to establish whether naloxone, an opioid receptor antagonist, abolishes the adaptation to ischemia observed in humans during coronary angioplasty after repeated balloon inflations. BACKGROUND: Experimental studies indicate that myocardial opioid receptors are involved in ischemic preconditioning. METHODS: Twenty patients undergoing angioplasty for an isolated stenosis of a major epicardial coronary artery were randomized to receive intravenous infusion of naloxone or placebo during the procedure. Intracoronary electrocardiogram and cardiac pain (using a 100-mm visual analog scale) were determined at the end of the first two balloon inflations. Average peak velocity in the contralateral coronary artery during balloon occlusion, an index of collateral recruitment, was also assessed by using a Doppler guide wire in the six patients of each group with a stenosis on the left anterior descending coronary artery. RESULTS: In naloxone-treated patients, ST-segment changes and cardiac pain severity during the second inflation were similar to those observed during the first inflation (12+/-6 vs. 11+/-7 mm, p = 0.3, and 58+/-13 vs. 56+/ 12 mm, p = 0.3, respectively), whereas in placebo-treated patients, they were significantly less (6+/-3 vs. 13+/-6 mm, p = 0.002 and 31+/-21 vs. 55+/-22 mm, p = 0.008, respectively). In both naloxone- and placebo-treated patients, average peak velocity significantly increased from baseline to the end of the first inflation (p = 0.04 and p = 0.02, respectively), but it did not show any further increase during the second inflation. CONCLUSIONS: The adaptation to ischemia observed in humans after two sequential coronary balloon inflations is abolished by naloxone and is independent of collateral recruitment. Thus, it is due to ischemic preconditioning and is, at least partially, mediated by opioid receptors, suggesting their presence in the human heart. PMID- 10362187 TI - Clinical validation of intravascular ultrasound imaging for assessment of coronary stenosis severity: comparison with stress myocardial perfusion imaging. AB - OBJECTIVES: To validate intravascular ultrasound (IVUS) measurements for differentiating functionally significant from nonsignificant coronary stenosis. BACKGROUND: To date, there are no validated criteria for the definition of a flow limiting coronary artery stenosis by IVUS. METHODS: Preinterventional IVUS imaging (30-MHz imaging catheter) of 70 de novo coronary lesions was performed. The lesion lumen area and three IVUS-derived stenosis indixes comparing lesion lumen area with the lesion external elastic lamina (EEL) area, the mean reference lumen area and the mean reference EEL area were compared with the results of stress myocardial perfusion imaging. RESULTS: The lesion lumen area and three IVUS-derived stenosis indexes showed sensitivities and specificities ranging between 80% and 90% using stress myocardial perfusion imaging as the gold standard. The lesion lumen area < or =4 mm2 is a simple and highly accurate criterion for significant coronary narrowing. CONCLUSIONS: Quantitative IVUS indices can be reliably used for identifying significant epicardial coronary artery stenoses. PMID- 10362189 TI - A comparison of the national registry of myocardial infarction 2 with the cooperative cardiovascular project. AB - OBJECTIVES: This study was performed to evaluate whether or not the simpler case identification and data abstraction processes used in National Registry of Myocardial Infarction two (NRMI 2) are comparable with the more rigorous processes utilized in the Cooperative Cardiovascular Project (CCP). BACKGROUND: The increased demand for quality of care and outcomes data in hospitalized patients has resulted in a proliferation of databases of varying quality. For patients admitted with myocardial infarction, there are two national databases that attempt to capture critical process and outcome data using different case identification and abstraction processes. METHODS: We compared case ascertainment and data elements collected in Medicare-eligible patients included in the industry-sponsored NRMI 2 with Medicare enrollees included in the Health Care Financing Administration-sponsored CCP who were admitted during identical enrollment periods. Internal and external validity of NRMI 2 was defined using the CCP as the "gold standard." RESULTS: Demographic and procedure use data obtained independently in each database were nearly identical. There was a tendency for NRMI 2 to identify past medical histories such as prior infarct (29% vs. 31%, p < 0.001) or heart failure (21% vs. 25%, p < 0.001) less frequently than the CCP. Hospital mortality was calculated to be higher in NRMI 2 (19.7% vs. 18.1%, p < 0.001) due mostly to the inclusion of noninsured patients 65 years and older in NRMI 2. CONCLUSIONS: We conclude that the simpler case ascertainment and data collection strategies employed by NRMI 2 result in process and outcome measures that are comparable to the more rigorous methods utilized by the CCP. Outcomes that are more difficult to measure from retrospective chart review such as stroke and recurrent myocardial infarction must be interpreted cautiously. PMID- 10362188 TI - A multicenter, randomized study of argatroban versus heparin as adjunct to tissue plasminogen activator (TPA) in acute myocardial infarction: myocardial infarction with novastan and TPA (MINT) study. AB - OBJECTIVES: This study examined the effect of a small-molecule, direct thrombin inhibitor, argatroban, on reperfusion induced by tissue plasminogen activator (TPA) in patients with acute myocardial infarction (AMI). BACKGROUND: Thrombin plays a crucial role in thrombosis and thrombolysis. In vitro and in vivo studies have shown that argatroban has advantages over heparin for the inhibition of clot bound thrombin and for the enhancement of thrombolysis with TPA. METHODS: One hundred and twenty-five patients with AMI within 6 h were randomized to heparin, low-dose argatroban or high-dose argatroban in addition to TPA. The primary end point was the rate of thrombolysis in myocardial infarction (TIMI) grade 3 flow at 90 min. RESULTS: TIMI grade 3 flow was achieved in 42.1% of heparin, 56.8% of low-dose argatroban (p = 0.20 vs. heparin) and 58.7% of high-dose argatroban patients (p = 0.13 vs. heparin). In patients presenting after 3 h, TIMI grade 3 flow was significantly more frequent in high-dose argatroban versus heparin patients: 57.1% versus 20.0% (p = 0.03 vs. heparin). Major bleeding was observed in 10.0% of heparin, and in 2.6% and 4.3% of low-dose and high-dose argatroban patients, respectively. The composite of death, recurrent myocardial infarction, cardiogenic shock or congestive heart failure, revascularization and recurrent ischemia at 30 days occurred in 37.5% of heparin, 32.0% of low-dose argatroban and 25.5% of high-dose argatroban patients (p = 0.23). CONCLUSIONS: Argatroban, as compared with heparin, appears to enhance reperfusion with TPA in patients with AMI, particularly in those patients with delayed presentation. The incidences of major bleeding and adverse clinical outcome were lower in the patients receiving argatroban. PMID- 10362190 TI - Prevalence, characteristics and prognostic value during long-term follow-up of nonsustained ventricular tachycardia after myocardial infarction in the thrombolytic era. AB - OBJECTIVES: The purpose of this study was to determine the prevalence, characteristics and the predictive value of nonsustained ventricular tachycardia (VT) for subsequent death and arrhythmic events after acute myocardial infarction (AMI). BACKGROUND: Nonsustained VT has been linked to an increased risk for sudden death in coronary patients. It is unknown whether this parameter can be used for selection of high-risk patients to receive an implantable defibrillator for primary prevention of sudden death in patients shortly after AMI. METHODS: In 325 consecutive infarct survivors, 24-h Holter monitoring was performed 10+/-6 days after AMI. All patients underwent coronary angiography, determination of left ventricular function and assessment of heart rate variability (HRV). Mean follow-up was 30+/-22 months. RESULTS: There was a low prevalence (9%) of nonsustained VT shortly after AMI. Nonsustained VT together with depressed left ventricular ejection fraction (LVEF) was found in only 2.4% of patients. During follow-up, 25 patients reached one of the prospectively defined end points (primary composite end point of cardiac death, sustained VT or resuscitated ventricular fibrillation; secondary end point: arrhythmic events). Kaplan Meier event probability analyses revealed that only HRV, LVEF and status of the infarct related artery were univariate predictors of death or arrhythmic events. The presence of nonsustained VT carried a relative risk of 2.6 for the primary study end point but was not a significant predictor if only arrhythmic events were considered. On multivariate analysis, only HRV, LVEF and the status of the infarct artery were found to be independently related to the primary study end point. CONCLUSIONS: There is a low prevalence of nonsustained VT shortly after AMI. Only 2% to 3% of all infarct survivors treated according to contemporary guidelines demonstrate both depressed LVEF and nonsustained VT. The predictive value of nonsustained VT for subsequent mortality and arrhythmic events is inferior to that of impaired autonomic tone, LVEF or infarct-related artery patency. Accordingly, the use of nonsustained VT to select patients for primary implantable cardioverter/defibrillator prevention trials shortly after AMI appears to be limited. PMID- 10362191 TI - Improved survival rates support left ventricular assist device implantation early after myocardial infarction. AB - OBJECTIVES: Implantation of left ventricular assist devices (LVADs) early after acute myocardial infarction (MI) has traditionally been thought to be associated with high mortality rates due to technical limitations and severe end-organ dysfunction. At some experienced centers, doctors have refrained from earlier operation after MI to allow for a period of hemodynamic and end-organ stabilization. METHODS: We retrospectively investigated the effect of preoperative MI on the survival rates of 25 patients who received a Thermocardiosystems Incorporated LVAD either <2 weeks (Early) (n = 15) or >2 weeks (Late) (n = 10) after MI. Outcome variables included perioperative right ventricular assistance (and right-sided circulatory failure), hemodynamic indexes, percent transplanted or explanted, and mortality. RESULTS: No statistically significant differences were demonstrated between demographic, perioperative or hemodynamic variables between the Early and Late groups. Patients in the Early group demonstrated a lower rate of perioperative mechanical right ventricular assistance, but had a higher rate of perioperative inhaled nitric oxide use. In addition, 67% of patients in the Early group survived to transplantation and 7% to explantation, findings comparable to those in the Late group (60% and 0% respectively). CONCLUSIONS: This clinical experience suggests that patients may have comparable outcomes whether implanted early or late after acute MI. These data therefore support the early identification and timely application of this modality in post-MI LVAD candidates, as this strategy may also reveal a subgroup of patients for whom post-MI temporary LVAD insertion may allow for full ventricular recovery. PMID- 10362192 TI - The significance of stress-induced ST segment depression in patients with inferior Q wave myocardial infarction. AB - OBJECTIVES: This study was conducted to evaluate the relationship between ST segment depression (STD) during dobutamine stress tests in different electrocardiogram (ECG) leads and myocardial ischemia assessed by simultaneous single photon emission computed tomography (SPECT) imaging in patients with inferior Q wave myocardial infarction. BACKGROUND: STD is a standard electrocardiographic sign of myocardial ischemia. Although STD may represent reciprocal changes in patients with previous myocardial infarction, studies of reciprocal changes during stress tests are scarce. METHODS: Dobutamine (up to 40 microg/kg/min) stress and rest myocardial perfusion scintigraphy using technetium SPECT imaging was performed in 125 patients >3 months after Q wave inferior myocardial infarction. The location of STD at the ECG was defined as anterior (V1 4), high lateral (I, aVL) and lateral (V5,6). Ischemia was defined as reversible perfusion abnormalities. RESULTS: STD occurred in the high lateral leads in 20 patients, in the anterior leads in 12 patients and in the lateral leads in 2 patients. ST segment elevation occurred in 25 patients in the inferior leads. High lateral STD was associated with inferior ST elevation in 16 patients (80%). There was a significant inverse linear correlation between the magnitude of ST segment shift from rest to peak stress in the inferior and the high lateral leads (r = -0.8, p < 0.0005), whereas no significant correlation was found between ST segment shift in the inferior and the anterior leads (r = -0.1, p = NS) or between the inferior and the lateral leads (r = 0.15, p = NS). Ischemia was detected in 45% of patients with and in 42% of patients without high lateral STD (p = NS). Patients with high lateral STD had a higher prevalence of fixed perfusion defects in the inferior wall (100% vs. 70%) and in the posterolateral wall (55% vs. 29%) compared with other patients (both p < 0.05). Ischemia was more prevalent in patients with anterior STD than without (75% vs. 39%, p < 0.05). CONCLUSIONS: In patients with inferior Q wave myocardial infarction, stress-induced STD in high lateral leads should be recognized as a reciprocal change for ST elevation in the inferior leads, and therefore, should be interpreted with the consideration of the significance of ST elevation if present, rather than being indicative of myocardial ischemia on its own. The STD found in the anterior leads appears to be a sign of myocardial ischemia. These findings should be considered in the definition of a positive ECG stress test and in establishing the criteria for the termination of stress test. PMID- 10362193 TI - Increased winter mortality from acute myocardial infarction and stroke: the effect of age. AB - OBJECTIVES: We examined seasonal variations in mortality from acute myocardial infarction (AMI) and stroke by age using 300,000 deaths in the Canadian Mortality Database for the years 1980 to 1982 and 1990 to 1992. BACKGROUND: The effect of age on environmental determinants of AMI and stroke is not well understood. METHODS: Seasonal variations were analyzed by month and for the four seasons (winter beginning in December). A chi-square test was used to test for homogeneity at p < 0.01, and relative risk ratios (RRs) for high and low periods were determined in relation to the overall mean. For each of four age subgroups, the magnitude of the seasonal variation was reported as the difference in mortality between the highest and lowest frequency seasons. RESULTS: By month, AMI deaths were highest in January (RR = 1.090) and lowest in September (RR = 0.904), a relative risk difference of 18.6%. The seasonal mortality variation in AMI deaths (winter vs. summer) increased with increasing age: 5.8% for <65, 8.3% for 65 to 74, 13.4% for 75 to 84 and 15.8% for >85 years (p < 0.005 for trend). Stroke mortality peaked in January (RR = 1.113) and had a trough in September (RR = 0.914), a relative risk difference of 19.9%. The seasonal variation in stroke mortality also increased with age. Seasonal variations were not seen in those aged <65 years, compared with 11.6% for 65 to 74, 15.2% for 75 to 84 and 19.3% for >85 years (p < 0.005 for trend). CONCLUSIONS: The elderly demonstrate a greater winter increase in AMI and stroke mortality than younger individuals. An understanding of these seasonal patterns may provide novel avenues for research in cardiovascular disease prevention. PMID- 10362194 TI - Aspirin and mortality in patients treated with angiotensin-converting enzyme inhibitors: a cohort study of 11,575 patients with coronary artery disease. AB - OBJECTIVES: The purpose of this study was to investigate the significance of the possible negative interaction between aspirin and angiotensin-converting enzyme (ACE) inhibitors. BACKGROUND: Several provocative reports have recently suggested that aspirin is unsafe in patients with heart failure and has negative interaction with ACE inhibitors that might attenuate their beneficial effects upon survival. METHODS: We analyzed mortality data of 11,575 patients with coronary artery disease screened for the Bezafibrate Infarction Prevention trial. A total of 1,247 patients (11%) were treated with ACE inhibitors. Of them, 618 patients (50%) used aspirin. RESULTS: Five-year mortality was lower among patients on ACE inhibitors and aspirin than patients on ACE inhibitors without aspirin (19% vs. 27%; p < 0.001). After adjusting for confounders, treatment with aspirin and ACE inhibitors remained associated with lower mortality risk than using ACE inhibitors only (relative risk [RR] = 0.71; 95% confidence interval [CI] = 0.56 to 0.91). Subgroup analysis of 464 patients with congestive heart failure treated with ACE inhibitors revealed 221 patients (48%) on aspirin and 243 patients not on aspirin. Although clinical characteristics and therapy were similar, patients taking aspirin experienced lower mortality than patients who did not (24% vs. 34%; p = 0.001). After adjustment, treatment with aspirin was still associated with lower mortality (RR = 0.70; 95% CI = 0.49 to 0.99). CONCLUSIONS: Among coronary artery disease patients with and without heart failure who are treated with ACE inhibitors, the use of aspirin was associated with lower mortality than treatment without aspirin. Our findings contradict the claim that aspirin attenuates the beneficial effect of ACE inhibitors and supports its use in patients with coronary artery disease treated with ACE inhibitors. PMID- 10362196 TI - Natural variability of circulating levels of cytokines and cytokine receptors in patients with heart failure: implications for clinical trials. AB - OBJECTIVES: The purpose of this study was to examine the variability in cytokines and cytokine receptors in patients with heart failure in comparison with a group of healthy control subjects who were free of cardiovascular disease. BACKGROUND: Despite increasing interest in cytokines as mediators of disease progression in heart failure and the recent interest in suppressing cytokines in clinical studies, the extent of variability in cytokines and cytokine receptors is largely unknown. This information is important for interpreting the results of studies in which changes in cytokine levels are measured in response to a specific form of therapy. METHODS: Circulating levels of tumor necrosis factor-alpha (TNF-alpha), and soluble TNF receptors (types 1 and 2), as well as interleukin (IL)-6 and IL-6 receptor were measured on a daily, weekly and monthly basis in heart failure patients (New York Heart Association class IIIa and IIIb; n = 10) and healthy volunteer subjects (n = 10). Measurements of cytokines and cytokine receptors were performed on plasma samples by enzyme-linked immunoassay. The daily, weekly and monthly degree of variability in cytokine and cytokine receptor levels was assessed by determining the coefficient of variation each point in time. RESULTS: The coefficient of variation for TNF-alpha and IL-6 levels increased over time in patients with heart failure; moreover, the coefficient of variation in heart failure subjects was significantly greater for IL-6 than for TNF-alpha. The coefficient of variation in cytokine receptor levels was minimal, and did not differ significantly between heart failure and control subjects. CONCLUSIONS: In patients with heart failure the degree of natural variability in circulating cytokine levels increases with time, and is greater for IL-6 than for TNF-alpha. Accordingly, the results of the present study suggest that the sample size needed to show a statistically significant change in the circulating level of a given cytokine will vary depending on the specific cytokine that is being measured, as well as the time period over which that cytokine is being assayed. PMID- 10362195 TI - Long-term effects of carvedilol in idiopathic dilated cardiomyopathy with persistent left ventricular dysfunction despite chronic metoprolol. The Heart Muscle Disease Study Group. AB - OBJECTIVES: The purpose of this study was to analyze whether long-term treatment with the nonselective beta-adrenergic blocking agent carvedilol may have beneficial effects in patients with dilated cardiomyopathy (DCM), who are poor responders in terms of left ventricular (LV) function and exercise tolerance to chronic treatment with the selective beta-blocker metoprolol. BACKGROUND: Although metoprolol has been proven to be beneficial in the majority of patients with heart failure, a subset of the remaining patients shows long-term survival without satisfactory clinical improvement. METHODS: Thirty consecutive DCM patients with persistent LV dysfunction (ejection fraction < or =40%) and reduced exercise tolerance (peak oxygen consumption <25 ml/kg/min) despite chronic (>1 year) tailored treatment with metoprolol and angiotensin-converting enzyme inhibitors were enrolled in a 12-month, open-label, parallel trial and were randomized either to continue on metoprolol (n = 16, mean dosage 142+/-44 mg/day) or to cross over to maximum tolerated dosage of carvedilol (n = 14, mean dosage 74+/-23 mg/day). RESULTS: At 12 months, patients on carvedilol, compared with those continuing on metoprolol, showed a decrease in LV dimensions (end-diastolic volume -8+/-7 vs. +7+/-6 ml/m2, p = 0.053; end-systolic volume -7+/-5 vs. +6+/-4 ml/m2, p = 0.047), an improvement in LV ejection fraction (+7+/-3% vs. -1+/-2%, p = 0.045), a reduction in ventricular ectopic beats (-12+/-9 vs. +62+/-50 n/h, p = 0.05) and couplets (-0.5+/-0.4 vs. +1.5+/-0.6 n/h, p = 0.048), no significant benefit on symptoms and quality of life and a negative effect on peak oxygen consumption (-0.6+/-0.6 vs. +1.3+/-0.5 ml/kg/min, p = 0.03). CONCLUSIONS: In DCM patients who were poor responders to chronic metoprolol, carvedilol treatment was associated with favorable effects on LV systolic function and remodeling as well as on ventricular arrhythmias, whereas it had a negative effect on peak oxygen consumption. PMID- 10362197 TI - Relationship between exertional symptoms and functional capacity in patients with heart failure. AB - OBJECTIVES: The present study was undertaken to investigate the relationship over time between exertional symptoms in heart failure and functional capacity. BACKGROUND: Most clinicians rely on exertional symptoms rather than on exercise testing to assess functional capacity in heart failure. However, it remains uncertain whether the subjective symptoms reported by patients provide a reliable index of functional capacity. METHODS: Fifty patients with heart failure underwent serial cardiopulmonary exercise testing and evaluation of exertional fatigue and dyspnea over a period of one to four years. Exercise testing was performed using the Naughton treadmill protocol and a MedGraphics metabolic cart. Fatigue and dyspnea were each scored from 0 to 3 (p = none, 1 = mild, 2 = moderate, 3 = severe). A composite symptom score was determined by adding together the fatigue and dyspnea scores. RESULTS: Patients underwent a total of 185 tests at an average interval of 4.3 months (average tests/patient = 3.7). Composite symptom scores noted at the time of exercise testing correlated significantly with peak exercise minute oxygen consumption (VO2) (r = 0.47, p < 0.01). In addition, the change in symptoms scores and change in peak VO2 noted between the baseline and final exercise test correlated significantly (r = 0.50, p < 0.01). However, patients reported few or no symptoms (symptom score < or =2) 45% of the time when peak VO2 was <14 ml/min/kg, consistent with a severe functional disability, and 72% of the time when peak VO2 was 14 to 18 ml/min/kg, consistent with moderate functional disability. CONCLUSIONS: Exertional symptoms reported by patients with heart failure generally correlate with maximal exercise capacity. However, exertional symptoms frequently underestimate the severity of functional disability. Cardiopulmonary exercise testing rather than symptoms should be used to assess functional capacity in heart failure. PMID- 10362198 TI - Congestive heart failure in subjects with normal versus reduced left ventricular ejection fraction: prevalence and mortality in a population-based cohort. AB - OBJECTIVES: The purpose of this study was to assess the relative proportions of normal versus impaired left ventricular (LV) systolic function among persons with congestive heart failure (CHF) in the community and to compare their long-term mortality during follow-up. BACKGROUND: Several hospital-based investigations have reported that a high proportion of subjects with CHF have normal LV systolic function. The prevalence and prognosis of CHF with normal LV systolic function in the community are not known. METHODS: We evaluated the echocardiograms of 73 Framingham Heart Study subjects with CHF (33 women, 40 men, mean age 73 years) and 146 age- and gender-matched control subjects (nested case-control study). Impaired LV systolic function was defined as an LV ejection fraction (LVEF) <0.50. RESULTS: Thirty-seven CHF cases (51%) had a normal LVEF; 36 (49%) had a reduced LVEF. Women predominated in the former group (65%), whereas men constituted 75% of the latter group. During a median follow-up of 6.2 years, CHF cases with normal LVEF experienced an annual mortality of 8.7% versus 3.0% for matched control subjects (adjusted hazards ratio = 4.06, 95% confidence interval 1.61 to 10.26). Congestive heart failure cases with reduced LVEF had an annual mortality of 18.9% versus 4.1% for matched control subjects (adjusted hazards ratio = 4.31, 95% confidence interval 1.98 to 9.36). CONCLUSIONS: Normal LV systolic function is often found in persons with CHF in the community and is more common in women than in men. Although CHF cases with normal LVEF have a lower mortality risk than cases with reduced LVEF, they have a fourfold mortality risk compared with control subjects who are free of CHF. PMID- 10362199 TI - Capillary density of skeletal muscle: a contributing mechanism for exercise intolerance in class II-III chronic heart failure independent of other peripheral alterations. AB - OBJECTIVES: The study was conducted to determine if the capillary density of skeletal muscle is a potential contributor to exercise intolerance in class II III chronic heart failure (CHF). BACKGROUND: Previous studies suggest that abnormalities in skeletal muscle histology, contractile protein content and enzymology contribute to exercise intolerance in CHF. METHODS: The present study examined skeletal muscle biopsies from 22 male patients with CHF compared with 10 age-matched normal male control patients. Aerobic capacities, myosin heavy chain (MHC) isoforms, enzymes, and capillary density were measured. RESULTS: The patients with CHF demonstrated a reduced peak oxygen consumption when compared to controls (15.0+/-2.5 vs. 19.8+/-5.0 ml x kg(-1) x min(-1), p <0.05). Using cell specific antibodies to directly assess vascular density, there was a reduction in capillary density in CHF measured as the number of endothelial cells/fiber (1.42+/-0.28 vs. 1.74+/-0.35, p = 0.02). In CHF, capillary density was inversely related to maximal oxygen consumption (r = 0.479, p = 0.02). The MHC IIx isoform was found to be higher in patients with CHF versus normal subjects (28.5+/-13.6 vs. 19.5+/-9.4, p <0.05). CONCLUSIONS: There was a significant reduction in microvascular density in patients with CHF compared with the control group, without major differences in other usual histologic and biochemical aerobic markers. The inverse relationship with peak oxygen consumption seen in the CHF group suggests that a reduction in microvascular density of skeletal muscle may precede other skeletal muscle alterations and play a critical role in the exercise intolerance characteristic of patients with CHF. PMID- 10362200 TI - Outcome of patients with nonischemic dilated cardiomyopathy and unexplained syncope treated with an implantable defibrillator. AB - OBJECTIVES: The purpose of this study was to determine the outcome of patients with nonischemic dilated cardiomyopathy, unexplained syncope and a negative electrophysiology test who are treated with an implantable defibrillator. BACKGROUND: Patients with nonischemic cardiomyopathy and unexplained syncope may be at high risk for sudden cardiac death, and they are sometimes treated with an implantable defibrillator. METHODS: This study prospectively determined the outcome of 14 consecutive patients who had a nonischemic cardiomyopathy, unexplained syncope and a negative electrophysiology test and who underwent defibrillator implantation (Syncope Group). Nineteen consecutive patients with a nonischemic cardiomyopathy and a cardiac arrest who were treated with a defibrillator (Arrest Group) served as a control group. RESULTS: Seven of 14 patients (50%) in the Syncope Group received appropriate shocks for ventricular arrhythmias during a mean follow-up of 24+/-13 months, compared with 8 of 19 patients (42%) in the Arrest Group during a mean follow-up of 45+/-40 months (p = 0.1). The mean duration from device implantation until the first appropriate shock was 32+/-7 months (95% confidence interval [CI], 18 to 45 months) in the Syncope Group compared to 72+/-12 months (95% CI, 48 to 96 months) in the Arrest Group (p = 0.1). Among patients who received appropriate shocks, the mean time from defibrillator implantation to the first appropriate shock was 10+/-14 months in the Syncope Group, compared with 48+/-47 months in the Arrest Group (p = 0.06). Recurrent syncope was always associated with ventricular tachyarrhythmias. CONCLUSIONS: The high incidence of appropriate defibrillator shocks and the association of recurrent syncope with ventricular arrhythmias support the treatment of patients with nonischemic cardiomyopathy, unexplained syncope and a negative electrophysiology test with an implantable defibrillator. PMID- 10362201 TI - Should ICDs be implanted in all patients with dilated cardiomyopathy and unexplained syncope? PMID- 10362202 TI - Implantable atrial defibrillator with a single-pass dual-electrode lead. AB - OBJECTIVES: We examined the feasibility and efficacy of using a single-pass, dual electrode (Solo) lead for atrial fibrillation (AF) detection and defibrillation. BACKGROUND: The efficacy and safety of an implantable atrial defibrillator (IAD) has been extensively studied; however, separate right atrial (RA) and coronary sinus (CS) defibrillation leads are used for the present system. METHODS: We studied the use of the Solo lead for AF detection and defibrillation in 17 patients who underwent cardioversion of chronic AF. The Solo lead with a proximal 6-cm RA electrode and a distal 6-cm spiral-shaped CS electrode were positioned into the CS with the RA electrode against the anterolateral RA wall. The RA-CS electrogram signal amplitudes were measured and the efficacy of the Solo lead for AF detection and defibrillation was assessed by using an external version of the IAD. RESULTS: The leads were inserted in all patients without complication (mean fluoroscopy time: 13.3+/-6.8 min). The mean RA-CS signal amplitude was 484+/-229 microV during sinus rhythm and 274+/-88 microV during AF (p < 0.05). All patients had satisfactory atrial signal amplitude to allow accurate detection of sinus rhythm. Successful cardioversion was achieved in 16/17 (94%) patients with an atrial defibrillation threshold of 320+/-70 V (5.5+/-2.7 J). Insufficient interelectrode spacing resulted in suboptimal electrode locations, associated with a lower atrial signal amplitude, a higher atrial defibrillation threshold and diaphragmatic stimulation. CONCLUSIONS: These results suggest a simplified lead configuration with optimal interelectrode spacing can be used with an IAD for AF detection and defibrillation. PMID- 10362203 TI - Evaluation of right atrial and biatrial temporary pacing for the prevention of atrial fibrillation after coronary artery bypass surgery. AB - OBJECTIVES: The purpose of this study was to determine if atrial pacing is effective in reducing postoperative atrial fibrillation (AF). BACKGROUND: Atrial fibrillation after coronary artery bypass grafting (CABG) is a common problem for which medical management has been disappointing. Atrial-based pacing has become an attractive nonpharmacologic therapy for the prevention of AF. METHODS: Sixty one post-CABG patients (mean age = 65 years) were randomized to one of three groups: no atrial pacing (NAP), right atrial pacing (RAP) or biatrial pacing (BAP). Each patient had one set of atrial wires attached to both the right and left atria, respectively, at the conclusion of surgery. Patients in the RAP and BAP groups were continuously paced at a rate of 100 pulses per minute for 96 h or until the onset of sustained AF (>10 min). All patients were monitored with Holter monitors or full disclosure telemetry to identify the onset of AF. The primary end point of the study was the first onset of sustained AF. RESULTS: There was no significant difference in the proportion of patients developing AF in the three groups (NAP = 33%; RAP = 29%; BAP = 37%; p > 0.7). However, for the subset of patients on beta-adrenergic blocking agents after CABG, there was a trend toward less AF in the paced groups. There were no serious complications related to pacing, although in three patients the pacemaker appeared to induce AF by pacing during atrial repolarization. CONCLUSIONS: Continuous right or biatrial pacing in the postoperative setting is safe and well tolerated. We did not find that post-CABG pacing prevented AF in this pilot study; however, the role of combined pacing and beta-blockade merits further study. PMID- 10362204 TI - Long-term reproducibility of electrophysiologically guided therapy with sotalol in patients with ventricular tachyarrhythmias. AB - OBJECTIVES: Goal of this study was to assess the long-term reproducibility of electrophysiologic drug testing in patients with ventricular tachyarrhythmias (VT/VF). BACKGROUND: Programmed ventricular stimulation (PVS) is still widely used to guide antiarrhythmic therapy in patients with sustained ventricular tachycardia/fibrillation (VT/VF). Sotalol is considered as one of the most effective drugs for VT/VF. Because there is no proof of long-term reproducibility of a successful drug test with sotalol, we investigated the long-term reproducibility of drug testing with sotalol. METHODS: Thirty patients with VT/VF (age: 57+/-11 years, 20 patients with coronary heart disease, 7 patients with no structural heart disease, 3 with others) and reproducible induction of VT/VF (28 patients VT, two patients VF) in a baseline PVS, were suppressible with sotalol (mean dosage 395+/-137 mg) in a subsequent PVS. After a mean follow-up of 13+/-10 months a PVS was again performed in patients, who had no evidence of progressive cardiac disease, who did not experience any arrhythmia recurrences or who were drug compliant. Irrespective of the inducibility after long-term therapy with sotalol, all patients were kept on the initial sotalol regimen. All 30 patients had a stable cardiac condition, were free of VT/VF recurrences and were drug compliant. RESULTS: Despite the clinical efficacy of sotalol, in 12 patients (40%) VT/VF could again be induced after 13+/-10.2 months. Inducibility was independent of age, heart disease, ejection fraction and follow-up time. During a further follow-up of 22.1+/-10.9 months, five patients experienced nonfatal VT recurrences independently of the prior inducibility. CONCLUSIONS: This study shows a lacking long-term reproducibility of an initial effective PVS with sotalol. Despite an uneventful clinical follow-up, late electrophysiologic testing showed a VT/VF inducibility in a high portion of patients. Hence, electrophysiologic testing performed late after the initial drug test may no longer be predictive of outcome. PMID- 10362205 TI - Partial cavotricuspid isthmus block before ablation in patients with typical atrial flutter. AB - OBJECTIVES: The purpose of this study was to prospectively evaluate preexisting partial isthmus block in the context of an electrophysiologically directed linear ablation strategy for typical atrial flutter (AF). BACKGROUND: Double potentials (DPs) separated by an isoelectric interval have been recognized as markers of local block. However, the presence and significance of DPs in the cavotricuspid isthmus during AF before ablation have not been evaluated. METHODS: Thirty consecutive patients with AF (counterclockwise: 24, clockwise: 6) were studied during AF. Sequential withdrawal mapping was performed in the cavotricuspid isthmus from the tricuspid valve (TV) to the inferior vena cava (IVC) edge with electrograms coinciding with the center of the surface electrocardiographic plateau during counterclockwise AF or with the initial downslope of the positive flutter wave during clockwise AF. Atrial electrograms along this line were categorized as double, single or fractionated potentials (SPs or FPs). After demarcation of the zone of contiguous DPs, radiofrequency (RF) catheter ablation was performed during AF only at sites with SPs or FPs (other than DPs) on the mapped line. If isthmus conduction still persisted after AF termination, additional RF applications were delivered using the same electrophysiologic strategy of avoiding DPs with an isoelectric interval during low lateral right atrial pacing for filling in the gap of residual conduction. RESULTS: Before ablation, no DPs were recorded in the isthmus in 19 patients (63%); DPs were recorded only at the IVC edge in five patients, and only at the TV edge in one patient. A contiguous line of DPs extending through more than half the isthmus to the IVC edge was documented in five patients (17%: group DP). In group DP, AF was terminated with 1.4+/-0.5 applications (vs. 5.8+/-3.5 in the remaining patients: p < 0.01). Complete isthmus block was achieved with a total of 3.4+/-0.5 applications (vs. 12+/-6 in the remaining patients: p < 0.01). CONCLUSIONS: Seventeen percent of patients undergoing ablation of AF have preexisting partial isthmus block indicated by a large contiguous zone of DPs separated by an isoelectric interval. Electrophysiologically directed linear ablation avoiding confluent DPs can prevent unnecessary applications for effective cure of AF. PMID- 10362206 TI - The natural history of aortic valve disease after mitral valve surgery. AB - OBJECTIVES: The present study evaluates the long-term course of aortic valve disease and the need for aortic valve surgery in patients with rheumatic mitral valve disease who underwent mitral valve surgery. BACKGROUND: Little is known about the natural history of aortic valve disease in patients undergoing mitral valve surgery for rheumatic mitral valve disease. In addition there is no firm policy regarding the appropriate treatment of mild aortic valve disease while replacing the mitral valve. METHODS: One-hundred thirty-one patients (44 male, 87 female; mean age 61+/-13 yr, range 35 to 89) were followed after mitral valve surgery for a mean period of 13+/-7 years. All patients had rheumatic heart disease. Aortic valve function was assessed preoperatively by cardiac catheterization and during follow-up by transthoracic echocardiography. RESULTS: At the time of mitral valve surgery, 59 patients (45%) had mild aortic valve disease: 7 (5%) aortic stenosis (AS), 58 (44%) aortic regurgitation (AR). At the end of follow-up, 96 patients (73%) had aortic valve disease: 33 AS (mild or moderate except in two cases) and 90 AR (mild or moderate except in one case). Among patients without aortic valve disease at the time of the mitral valve surgery, only three patients developed significant aortic valve disease after 25 years of follow-up procedures. Disease progression was noted in three of the seven patients with AS (2 to severe) and in six of the fifty eight with AR (1 to severe). Fifty two (90%) with mild AR remained stable after a mean follow-up period of 16 years. In only three patients (2%) the aortic valve disease progressed significantly after 9, 17 and 22 years. In only six patients of the entire cohort (5%), aortic valve replacement was needed after a mean period of 21 years (range 15 to 33). In four of them the primary indication for the second surgery was dysfunction of the prosthetic mitral valve. CONCLUSIONS: Our findings indicate that, among patients with rheumatic heart disease, a considerable number of patients have mild aortic valve disease at the time of mitral valve surgery. Yet most do not progress to severe disease, and aortic valve replacement is rarely needed after a long follow-up period. Thus, prophylactic valve replacement is not indicated in these cases. PMID- 10362207 TI - "Prophylactic" valve replacement for mild aortic valve disease at time of surgery for other cardiovascular disease?...No. PMID- 10362208 TI - The Mitral Regurgitation Index: an echocardiographic guide to severity. AB - OBJECTIVES: The purpose of this study was to develop a semiquantitative index of mitral regurgitation severity suitable for use in daily clinical practice and research. BACKGROUND: There is no simple method for quantification of mitral regurgitation (MR). The MR Index is a semiquantitative guide to MR severity. The MR Index is a composite of six echocardiographic variables: color Doppler regurgitant jet penetration and proximal isovelocity surface area, continuous wave Doppler characteristics of the regurgitant jet and tricuspid regurgitant jet derived pulmonary artery pressure, pulse wave Doppler pulmonary venous flow pattern and two-dimensional echocardiographic estimation of left atrial size. METHODS: Consecutive patients (n = 103) with varying grades of MR, seen in the Adult Echocardiography Laboratory at UCSF, were analyzed retrospectively. All patients were evaluated for the six variables, each variable being scored on a four point scale from 0 to 3. The reference standards for MR were qualitative echocardiographic evaluation by an expert and quantitation of regurgitant fraction using two-dimensional and Doppler echocardiography. A subgroup of patients with low ejection fraction (EF < 50%) were also analyzed. RESULTS: The MR Index increased in proportion to MR severity with a significant difference among the three grades in both normal and low EF groups (F = 130 and F = 42, respectively, p < 0.0001). The MR Index correlated with regurgitant fraction (r = 0.76, p < 0.0001). An MR Index > or =2.2 identified 26/29 patients with severe MR (sensitivity = 90%, specificity = 88%, PPV = 79%). No patient with severe MR had an MR Index <1.8 and no patient with mild MR had an MR Index >1.7. CONCLUSIONS: The MR Index is a simple semiquantitative estimate of MR severity, which seems to be useful in evaluating MR in patients with a low EF. PMID- 10362209 TI - Value and limitations of the Duke criteria for the diagnosis of infective endocarditis. AB - OBJECTIVES: The purpose of this study was to assess the value and limitations of Duke criteria for the diagnosis of infective endocarditis (IE). BACKGROUND: Duke criteria have been shown to be more sensitive in diagnosing IE than the von Reyn criteria, but the diagnosis of IE remains difficult in some patients. METHODS: Both classifications were applied in 93 consecutive patients with pathologically proven IE. Blood cultures, and transthoracic and transesophageal echocardiography were performed in all patients. RESULTS: Sensitivities for the diagnosis of IE were 56% and 76% for von Reyn and Duke criteria, respectively. Fifty-two patients were correctly classified as "probable IE" by von Reyn and "definite IE" by Duke criteria (group 1). However, discrepancies were observed in 41 patients. Eleven patients (group 2) were misclassified as "rejected" by von Reyn, but were "definite IE" by Duke criteria; this difference could be explained by negative blood cultures and positive echocardiogram in all patients. In eight patients (group 3), the diagnosis of IE was "possible" by von Reyn but "definite" by Duke criteria. This difference was essentially explained by the failure of the von Reyn classification to consider echocardiographic abnormalities as major criteria. Twenty-two patients (group 4) were misclassified as possible IE using Duke criteria, being false negative of this classification. Echocardiographic major criteria were present in 19 patients, but blood cultures were negative in 21 patients. The cause of negative blood cultures was prior antibiotic therapy in 11 patients and Q-fever endocarditis diagnosed by positive serology in three cases. CONCLUSIONS: Twenty-four percent of patients with proved IE remain misclassified as "possible IE" despite the use of Duke criteria, especially in cases of culture-negative and Q-fever IE. Increasing the diagnostic value of echographic criteria in patients with prior antibiotic therapy and typical echocardiographic findings and considering the serologic diagnosis of Q fever as a major criterion would further improve the clinical diagnosis of IE. PMID- 10362210 TI - Individual and combined effects of estrogen/progestin therapy and lovastatin on lipids and flow-mediated vasodilation in postmenopausal women with coronary artery disease. AB - OBJECTIVES: We sought to examine the individual and combined effects of estrogen/progestin therapy versus lovastatin on lipids and flow-mediated vasodilation in postmenopausal women with heart disease. BACKGROUND: Little information is available regarding the relative benefits of estrogen replacement therapy versus reductase inhibitors and the potential utility of their combination as lipid-lowering therapy for postmenopausal women. METHODS: We conducted a randomized, double-blind, crossover trial in 24 postmenopausal women, each of whom received the following drug regimens during three consecutive six week treatment periods: 1) hormone replacement (oral dose of 0.625 mg/day conjugated equine estrogens and 2.5 mg/day medroxyprogesterone acetate); 2) 20 mg lovastatin/day and 3) hormone replacement plus lovastatin. RESULTS: Total and low density lipoprotein (LDL) cholesterol were significantly lowered and high density lipoprotein (HDL) cholesterol was significantly increased by all three regimens compared with baseline (p < 0.05). Lovastatin was more effective than estrogen/progestin in reducing LDL (p < 0.001), but estrogen/progestin was slightly more effective in increasing HDL. The hormone replacement and lovastatin regimen blocked the estrogen-associated increase in triglycerides. Hormone replacement (alone and with lovastatin) resulted in increases in brachial artery flow-mediated vasodilator capacity (p = 0.01 for both regimens) and the area under the curve (p = 0.016 and p = 0.005, respectively) compared with baseline. Percent dilation was greatest after the hormone replacement regimen, whereas the area under the curve was greatest after hormone replacement plus lovastatin (69% improvement vs. baseline). CONCLUSIONS: In postmenopausal women with coronary disease and hyperlipidemia, conjugated equine estrogen produced significant improvements in lipids and vasodilator responses despite the concurrent administration of low dose medroxyprogesterone acetate. Low dose lovastatin produced greater reductions in LDL, but less dramatic improvements in vasodilator responses. Estrogen/progestin plus lovastatin may provide additional benefits via a greater reduction in the LDL/HDL ratio and attenuation of estrogen-associated hypertriglyceridemia. More information is needed about the safety and efficacy of such combinations of hormone replacement and reductase inhibitor therapy. PMID- 10362211 TI - Impaired brachial artery endothelial function is not predicted by elevated triglycerides. AB - OBJECTIVES: The purpose of this study was to determine if patients with modest hyperlipidemia, and no other risk factors for coronary artery disease (CAD), have impaired endothelium-dependent (ED) vasoactivity. BACKGROUND: Hypercholesterolemia impairs ED vasodilation, but the impact of elevated triglycerides on endothelial function is not as well established. METHODS: High resolution ultrasound was used to determine flow-mediated dilation (FMD) in the brachial artery (BA) after a 5-min arterial occlusion (endothelium-dependent stimulus) and nitroglycerin-induced dilation (endothelium-independent stimulus). We studied 40 healthy controls (Group 1), 38 patients with elevated low-density lipoprotein (LDL) cholesterol (Group 2) and 35 patients with elevated triglycerides (Group 3). Patients were excluded if they had known CAD or other risk factors for CAD, or if they were receiving lipid-lowering or vasoactive medications. RESULTS: Control patients (Group 1) had normal LDL cholesterol (2.6+/-0.8 mmol/liter) and triglyceride levels (1.0+/-0.5 mmol/liter) compared with Group 2 (5.2+/-1.2 mmol/liter, 1.8+/-0.6 mmol/liter) and Group 3 (3.5+/-0.9 mmol/liter, 4.2+/-2.5 mmol/liter) subjects (p < 0.001). Baseline BA diameters were the same across the three groups. There was no significant attenuation of flow-mediated vasodilation (FMD) in either of the hyperlipidemic groups (Group 1: 10.9+/-5.0% vs. Group 2: 8.6+/-6.1% vs. Group 3: 9.4+/-3.9%; p = 0.14). However, nitroglycerin-induced vasodilation was mildly reduced (Group 1: 21.0+/-5.0% vs. 16.9+/-7.6% vs. 17.3+/-7.7%; p = 0.01). By multivariate analysis, after controlling for baseline diameters, only the ratio of LDL/high-density lipoprotein predicted a minor impairment in FMD. CONCLUSIONS: In patients free from other cardiac risk factors, modest elevations of triglycerides or LDL cholesterol do not significantly attenuate BA endothelial-dependent vasodilation. Synergism with other cardiac risk factors may be required to significantly impair endothelial function in these patients. PMID- 10362212 TI - Prognostic value of systemic blood pressure response during exercise in a community-based patient population with hypertrophic cardiomyopathy. AB - OBJECTIVES: The present study was designed to prospectively evaluate the prognostic relevance of abnormal blood pressure response to exercise (ABPR), defined as hypotension or failed blood pressure increase (<20 mm Hg) with exercise, in a community-based hypertrophic cardiomyopathy (HCM) population representative of the overall disease spectrum. BACKGROUND: Abnormal blood pressure response to exercise has been proposed as a marker for hemodynamic instability and increased risk for disease-related mortality in highly selected patient populations with HCM. METHODS: The study population comprised 126 patients (aged 42+/-14 years) who underwent maximal symptom-limited cycloergometer exercise testing as part of the standard evaluation at our institution, and who were followed systematically for 4.7+/-3.7 years after testing. RESULTS: Of the 126 study patients, 98 (78%) had a normal blood pressure response during exercise, whereas the other 28 (22%) had ABPR, including nine with hypotension and 19 with failed blood pressure rise. During the follow-up period, nine patients (7%) died of HCM-related causes (three suddenly and six heart failure-related), of whom four had ABPR. In those patients aged < or =50 years, survival analysis after exercise testing showed a significantly increased risk for cardiovascular mortality associated with ABPR compared with a normal exercise response (p = 0.04), with an odds ratio of 4.5 (95% confidence interval: 1.1, 20.1). However, ABPR showed low positive predictive accuracy for cardiovascular mortality (i.e., 14%), whereas negative predictive accuracy was high (i.e., 95%). CONCLUSIONS: A hypotensive blood pressure response during exercise occurred in over 20% of a community-based patient cohort with HCM, and was associated with adverse long-term prognosis in patients <50 years old. However, the positive predictive accuracy of this blood pressure response is too low to justify modifications of clinical management or to allow identification of the high-risk patient based solely on an abnormal test result. By virtue of its high negative predictive accuracy for HCM-related mortality, the blood pressure response to exercise appears to be most valuable (in conjunction with the absence of other well recognized risk factors) as a screening test for the identification of low-risk subsets of patients. PMID- 10362213 TI - Bubble contrast echocardiography in detecting pulmonary arteriovenous shunting in children with univentricular heart after cavopulmonary anastomosis. AB - OBJECTIVES: We sought to compare bubble contrast echocardiography and pulmonary angiography in detecting pulmonary arteriovenous malformation (PAVM) in children with cavopulmonary anastomosis (CPA), and to examine anatomic and physiologic variables associated with the development of PAVM. BACKGROUND: Development of PAVM in patients with CPA may cause profound cyanosis. Pulmonary arteriovenous malformation has been traditionally diagnosed by pulmonary angiography with reported incidence of 20% to 25% in patients with CPA. METHODS: Fourteen patients (age 1.1 to 12.6 years) with any forms of CPA and normal pulmonary venous drainage formed the study population. All patients underwent cardiac catheterization and pulmonary angiography. Bubble contrast echocardiographic studies were performed with injection of 10 ml of agitated saline solution into branch pulmonary arteries. Transthoracic echocardiograms using an apical view were performed to assess the appearance of bubble contrast in the systemic ventricles. We compared the results of pulmonary angiograms and contrast echocardiograms, and findings of contrast echocardiograms between lungs with hepatic venous blood flow and lungs without hepatic venous blood. RESULTS: Ten of the 14 patients (71%) had positive contrast echocardiographic studies, compared with three (21%) detected by pulmonary angiograms (p = 0.01). No difference was found in pulmonary artery pressure, transpulmonary gradient or presence of heterotaxy syndrome between patients with positive contrast echocardiographic studies and patients with negative studies. However, patients with positive contrast echocardiograms tended to have lower oxygen saturation (81%) and higher hemoglobin (16.4 g/dl) compared with patients with negative studies (88% and 14.7 g/dl, p = 0.10 and p = 0.18 respectively). Patients with Glenn shunt or unidirectional Fontan had higher incidence of PAVM (10/11) compared with patients with classic or lateral tunnel Fontan (0/3, p = 0.01). All 12 lungs with no perfusion of hepatic venous blood had positive contrast echocardiographic studies. Lungs with no hepatic venous blood flow were more likely to develop PAVM compared with lungs with hepatic venous blood flow (12/12 and 3/16 respectively, p < 0.01). CONCLUSIONS: Bubble contrast echocardiography is more sensitive in detecting PAVM compared with pulmonary angiography. The prevalence of PAVM in patients with CPA may be much higher than what had been reported previously. Lungs with no hepatic venous blood flow are more likely to develop PAVM than lungs with hepatic venous blood flow. PMID- 10362214 TI - Neurally mediated cardiac syncope: autonomic modulation after normal saline infusion. AB - OBJECTIVES: This study assessed the heart variability response to orthostatic stress during tilt table testing before and after normal saline administration. BACKGROUND: The efficacy of sodium chloride and mineralocortoid in the treatment of neurally mediated cardiac syncope is attributed to intravascular volume expansion; however, their modulation of autonomic nervous system activity has not been evaluated. METHODS: Heart rate variability analysis was performed on 12 adolescents with a history of syncope or presyncope (mean age 15.2+/-0.7 years) during tilt table testing. Subjects were upright 80 degrees for 30 min or until syncope. After normal saline administration, the patient was returned upright for 30 min. Heart rate variability analysis data were analyzed by an autoregression model (Burg method). RESULTS: All subjects reproducibly developed syncope during control tilt table testing; median time to syncope was 9.4+/-2.1 min. After normal saline infusion, none of the subjects developed syncope after 30 min upright. In the control tilt, there was an initial increase followed by a progressive decrease in low frequency power until syncope. Repeat tilt after normal saline administration demonstrates that low frequency power increased but the magnitude of initial change was blunted when compared with control. In addition, low frequency power increased during normal saline tilt sequence compared with the control tilt, during which it decreased. CONCLUSIONS: Normal saline blunted low frequency power stimulation and prevented paradoxical low frequency power (sympathetic) withdrawal. Increasing intravascular volume with normal saline alters autonomic responses that may trigger neurally mediated syncope reflexes. PMID- 10362215 TI - Idiopathic ventricular tachycardia in infancy and childhood: a multicenter study on clinical profile and outcome. Working Group on Dysrhythmias and Electrophysiology of the Association for European Pediatric Cardiology. AB - OBJECTIVES: The present study intended to evaluate the clinical profile and outcome in a large cohort of pediatric patients with idiopathic ventricular tachycardia (VT). BACKGROUND: Ventricular tachycardia (VT) without underlying heart disease is rare in childhood. Limited information is available with regard to outcome and indications for long-term antiarrhythmic treatment. METHODS: A retrospective multicenter study was conducted. Patient data were obtained from the individual centers using a standardized questionnaire. RESULTS: Ninety-eight pediatric patients with episodes of VT in the absence of structural heart disease were included. Mean age at first manifestation of the arrhythmia was 5.4 years (range 0.1 to 15.1), with 27% of the patients having had VT already in infancy. Clinical symptoms or echocardiographic signs of left ventricular dysfunction were observed initially in 36% of the patients, of which one third (12% of the whole population) presented with severe symptoms (heart failure or syncope). After a mean follow-up of 47 months (range 12 to 182), no patient had died. Twenty-five patients had never been treated with antiarrhythmic drugs. Sixty-three patients were free of VT and did not take antiarrhythmic drugs at last follow-up. Prognosis was better when VT occurred during the first year of life (VT resolution in 89%) compared with VT occurrence beyond the first year of life (VT resolution in 56%: p < 0.01). The clinical profile was more favorable for patients with presumed right VT (VT resolution in 76%, symptoms in 25% of patients) compared with patients with presumed left VT, where VT resolution occurred in 37% and symptoms in 67% of patients (p < 0.01). CONCLUSIONS: VT in children with a normal heart carried a good prognosis. Outcome was better after onset of VT during infancy and when VT originated in the right ventricle. A restrictive use of antiarrhythmic agents might be justified in a large proportion of these patients. PMID- 10362217 TI - Lipid-lowering therapy for the primary prevention of coronary heart disease. PMID- 10362216 TI - Persistent primary coronary dilation induced by transatrial delivery of nitroglycerin into the pericardial space: a novel approach for local cardiac drug delivery. AB - OBJECTIVES: We compared the effects of intrapericardial and intracoronary nitroglycerin on coronary cross-sectional area as assessed by intravascular ultrasound and demonstrated the feasibility of local cardiac drug delivery by a newly developed method to access the normal pericardial space through the right atrial appendage. BACKGROUND: Studies of nitric oxide (NO) donors have suggested that their antiarrhythmic and antiproliferative properties are more effective when administered by the intrapericardial rather than intravascular route. We postulated that NO donors delivered intrapericardially would also cause sustained coronary vasodilation without significant systemic hypotension. METHODS: Intrapericardial nitroglycerin (200 microg) was administered in five Yorkshire pigs. Coronary cross-sectional luminal area was measured with intravascular ultrasound at various time intervals. The effects of intracoronary nitroglycerin on coronary luminal area were used for comparison. RESULTS: Transatrial pericardial access required 1 to 3 min in all animals. Intrapericardial nitroglycerin was associated with a mean 31.7% increase in luminal area at 5 min (p < 0.001). Vasodilation peaked between 5 and 10 min and persisted for 15 min. In contrast, intracoronary nitroglycerin was associated with a smaller mean increase in luminal area (20.3% at 5 min, p < 0.01) that disappeared by 10 min. Significant systemic hypotension was observed at 3 min with intracoronary but not with intrapericardial nitroglycerin. CONCLUSIONS: Sustained coronary vasodilation can be achieved with intrapericardial delivery of nitroglycerin without systemic hypotension. Nitric oxide donors with longer half-lives could prove beneficial in the treatment of myocardial ischemic syndromes when administered through this route. Transatrial pericardial access offers a novel route for local cardiac drug delivery. PMID- 10362218 TI - Editorial independence--what did we learn from the Journal of the American Medical Association? PMID- 10362219 TI - President's page: the ACC and subspecialty societies: the beauty of the quilt. PMID- 10362220 TI - Rescue angioplasty after failed thrombolysis--high mortality despite successful procedure. PMID- 10362221 TI - Transfer delay for primary PTCA: does it influence clinical outcome? Percutaneous transluminal coronary angioplasty. PMID- 10362222 TI - Seasonal distribution of myocardial infarction and seasonal mood changes. PMID- 10362223 TI - Left ventricular hypertrophy and sudden death. PMID- 10362224 TI - To trust or not to trust. PMID- 10362225 TI - ACC/AHA/ACP-ASIM guidelines for the management of patients with chronic stable angina: a report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines (Committee on Management of Patients With Chronic Stable Angina). PMID- 10362226 TI - Serum paraoxonase activity and phenotype distribution in Turkish subjects with coronary heart disease and its relationship to serum lipids and lipoproteins. AB - Recently, biochemical studies of paraoxonase in the serum of humans have shown that much of this enzymes' activity is associated with high-density lipoprotein (HDL) and paraoxonase may play a role in lipid metabolism preventing the accumulation of the lipoperoxides. In this study, a possible relationship between coronary heart disease (CHD) and paraoxonase activity were investigated. Serum triglycerides, total cholesterol, HDL-cholesterol and paraoxonase activity were measured in unrelated healthy donors and CHD patients. It was found that paraoxonase activity was trimodally distributed in both groups but no statistically significant difference was found between phenotype distributions of controls and CHD patients (gene frequencies; 0.632 and 0.382 of controls, 0.702 and 0.298 of patients for the A and B alleles, respectively). However, in CHD group, a high possibility was found to be phenotype A compared with the control group. A relative risk of 1.48 (95% confidence intervals (CI), 0.986-2.227) was found for the relation between CHD and the paraoxonase activity. Patients' HDL cholesterol values were lower and triglycerides were higher than controls (P<0.001). It may be concluded from the present study that although no statistically significant difference was found between paraoxonase phenotype distributions of controls and CHD patients, a decrease in paraoxonase activity could become a risk factor for this disease. PMID- 10362227 TI - Quinolinic acid, alpha-picolinic acid, fusaric acid, and 2,6-pyridinedicarboxylic acid enhance the Fenton reaction in phosphate buffer. AB - Quinolinic acid, alpha-picolinic acid, fusaric acid, and 2,6-pyridinedicarboxylic acid enhanced the Fenton reaction in phosphate buffer, respectively. The enhancement by quinolinic acid, alpha-picolinic acid, fusaric acid, and 2,6 pyridinedicarboxylic acid of the Fenton reaction may be partly related to their respective actions in the biological systems such as a neurotoxic effect (quinolinic acid), a marked growth-inhibitory action on rice seeding (alpha picolinic acid and fusaric acid), and an antiseptic (2,6-pyridinedicarboxylic acid). The ultraviolet-visible absorption spectrum of the mixture of alpha picolinic acid with ferrous ion showed a characteristic visible absorbance band with a lambda(max) at 443 nm, suggesting that alpha-picolinic acid chelate of Fe2+ ion forms in the solution. Similar characteristic visible absorbance band was also observed for the mixture of Fe2+ ion with quinolinic acid (or fusaric acid, or 2,6-pyridinedicarboxylic acid). The chelation seems to be related to the enhancement by quinolinic acid, alpha-picolinic acid, fusaric acid, and 2,6 pyridinedicarboxylic acid of the Fenton reaction. alpha-Picolinic acid was reported to be a toxic substance isolated from the culture liquids of blast mould (Piricularia oryzae CAVARA). On the other hand, it has also been known that chlorogenic acid protects rice plants from the blast disease. The chlorogenic acid inhibited the formation of the hydroxyl radical in the reaction mixture of alpha-picolinic acid, FeSO4(NH4)2SO4, and H2O2. Thus the inhibition may be a possible mechanism of the protective action of the chlorogenic acid against the blast disease. PMID- 10362228 TI - Influence of beta-adrenergic antagonists on cell proliferation rates in the kidney of untreated and diethylnitrosamine-treated male F344 rats. AB - Some nongenotoxic chemicals which cause kidney tumors have been shown to stimulate tubular cell proliferation. The aim of this study was to evaluate the effects of two beta-adrenoreceptor blocking agents, propranolol and atenolol, on cell proliferation rates in the kidneys of male F344 rats. Immunohistochemical expression of proliferating cell nuclear antigen (PCNA) and mitotic index have been examined in formalin-stored kidneys from F344 rats used in an initiation promotion study of carcinogenesis. Cell proliferation rate was quantified in the proximal tubule epithelium. Non-initiated rats and rats initiated with a single dose of diethylnitrosamine (DEN, 200 mg/kg, i.p.) were continuously treated with propranolol (75-100 mg/kg) or atenolol (300 mg/kg) by gavage and were sacrificed after 2, 4, 8 or 21 months of experimentation. There were two control groups, one untreated (D1) and one given distilled water by gavage (D1). Control group D1 showed significantly lower cell proliferation rates than the D0 group. In non initiated rats, propranolol had a weak enhancing effect on cell proliferation, most evident after 4 months, while atenolol had a clear enhancing effect most evident after 8 months of promoting regimen. Treatment with DENalone resulted in a significant increase in cell proliferation rate as compared to group D1. In DEN initiated rats given propranolol, there was a borderline significant increase in cell proliferation rates, compared to rats given DEN alone, after 8 months of promoting regimen. Atenolol had no effect. Because of the differences in body weight gain and food consumption observed among the various groups, it is suggested that the state of nutrition may have obscured the effects of beta blockers on cell proliferation rates. PMID- 10362229 TI - Characterization of a renal medium chain acyl-CoA synthetase responsible for glycine conjugation in mouse kidney mitochondria. AB - Glycine conjugation of a series of benzoic acid derivatives was investigated in mouse kidney mitochondria. The chlorine and methyl substitutions in the para- and meta-positions of the benzene ring yielded an increase in glycine conjugation. The acids with a methoxy group showed a low degree of glycine conjugation. In addition, the acids with nitro or amino groups were conjugated to a slight extent with glycine. The in vitro conjugation of salicylic acid with glycine occurred not in liver but in kidney. The specificity of the renal medium chain acyl-CoA synthetase catalyzing the first reaction of glycine conjugation was also examined. The enzyme accepted not only medium chain fatty acids but also aromatic and arylacetic acids. The highest activity was shown with hexanoic acid. High activities were observed for benzoic acid derivatives with alkyl and alkoxyl groups in the para- and meta-positions of the benzene ring. An ortho-substituted acid exhibited no activity. In addition, the enzyme was less active with valproic acid, tranexamic acid, indomethacin and ketoprofen. The enzyme was inhibited by diflunisal, 2-hydroxydodecanoic acid and salicylic acid, which did not act as substrates. There was a poor correlation between the activity of the medium chain acyl-CoA synthetase and glycine conjugation of eleven substituted benzoic acids. These findings suggest that the present medium chain acyl-CoA synthetase is involved in glycine conjugation of the substituted acids in mouse kidney mitochondria, but there may be a larger contribution of another isoenzyme. PMID- 10362230 TI - The profile of urinary metabolites of 4-(methylnitrosamino)-1-(3-pyridyl)-1 butanone in rats is determined by its pulmonary metabolism. AB - Metabolism of the tobacco specific nitrosamine 4-(methylnitrosamino)-1-(3 pyridyl)-1-butanone (NNK) in rats was compared to metabolism in primary lung and liver cells. Untreated rats and rats pretreated with phenobarbital, acetone or phenethyl isothiocyanate (PEITC) were used for all experiments. Also the influence of [-]-1-methyl-2-[3-pyridyl]-pyrrolidine (nicotine) administered concomitantly with NNK, or incubated with isolated cells, upon NNK metabolism was investigated and found to be only marginal upon alpha-hydroxylation and pyridine N-oxidation in vivo. In hepatocytes nicotine inhibited NNK pyridine N-oxidation, alpha-hydroxylation and glucuronidation of 4-(methylnitrosamino)-1-(3-pyridyl)-1 butanol (NNAL), whereas in lung cells the influence of nicotine was not as pronounced. In vivo phenobarbital induced alpha-hydroxylation and pyridine N oxidation. In vitro the effects of the modulators were most pronounced upon hepatocytes, where phenobarbital greatly induced pyridine N-oxidation and PEITC inhibited alpha-hydroxylation. NNAL was conjugated to its beta-glucuronide in lung cells at four times higher rates than in hepatocytes. The ratios of the sum of N-oxides to the sum of alpha-hydroxylation products in vivo were similar to those in lung cells, especially at low NNK concentrations (1 microM), while in hepatocytes alpha-hydroxylation was more pronounced. The same correlation of metabolism in isolated lung cells with whole rats was observed if oxidative NNAL metabolism was related to oxidative NNK metabolism. Here hepatocytes showed a much higher formation of NNAL oxidation products than either lung cells formed, or rats excreted in urine. This was true despite a lower rate of metabolism in the lung than in liver if based on cell number, the rate based on mg protein was four times higher in lung than liver. Only after phenobarbital treatment was the contribution of hepatic metabolism to excreted metabolites important. In conclusion the lung which is also the target of NNK carcinogenesis, and not the liver, is the organ with the most important contribution to NNK and NNAL metabolism at concentrations relevant to human exposure. PMID- 10362231 TI - Monoarticular synovial lesions: radiologic pictorial essay with pathologic illustration. AB - A wide variety of common focal monoarticular synovial lesions may be encountered on imaging studies. A multi-modality approach to these lesions, with appreciation of the differing appearances, can often yield the correct diagnosis. This pictorial essay demonstrates and describes the imaging with illustration of pathologic findings in a spectrum of conditions. PMID- 10362232 TI - The effect of hormone replacement therapy on the sensitivity of screening mammograms. AB - AIM: The use of hormone replacement therapy (HRT) can lead to various changes on the mammogram including increasing density. The object of this study was to assess the effect of HRT on the sensitivity of mammographic screening by comparing HRT usage in women with screen detected breast cancers with HRT usage at the time of screening in women presenting with interval cancers. METHODS: The West of Scotland Breast Screening Programme serves a population of 180,000 women aged 50-64 years old. Between May 1988 and December 1995, 1461 breast cancers were detected by the screening programme in 1441 women over the age of 50 and 372 interval breast cancers presented in 371 women screened between these dates. HRT usage at the time of screening was noted with details of age, postcode and the time between screening and diagnosis in the case of the women with interval cancers. RESULTS: Among women under 65 years old, screened between 1988-1993, 12.3% of women with screen detected cancers and 22.2% of women with interval cancers were using HRT (P<0.001). Further analysis demonstrates that interval cancer rate is related to age as well as HRT use. After adjusting for age at time of screening, deprivation category and year of screening, the relative risk of a woman using HRT having an interval cancer compared with that of a woman not using HRT is 1.79. The relative risk of an interval cancer arising in the first year after screening for a woman on HRT is 2.27. CONCLUSION: The use of HRT and being of an age below 60 years are both risk factors for presenting with an interval cancer after mammographic breast screening. Our results indicate that the use of HRT leads to a decrease in the sensitivity of mammographic screening. PMID- 10362233 TI - Palatal lymphoepitheliomas and a review of head and neck lymphoepitheliomas. AB - AIM: Lymphoepithelioma is principally a tumour of the nasopharynx with only sporadic cases arising elsewhere in the head and neck. We describe the clinical and imaging features of a group of rare lymphoepitheliomas related to the palate. PATIENTS AND METHODS: Four patients with lymphoepithelioma of the palate are described. In each case we retrospectively reviewed the clinical records, laboratory results, and imaging which consisted of computed tomography (CT) and ultrasound in all four cases and magnetic resonance imaging (MRI) in two patients. RESULTS: All four patients were ethnic Chinese (non-smokers, non drinkers). All cases were Epstein-Barr virus (EBV) related. Tumour was related to the palate in two cases and extended into the nasal cavity in one patient. The fourth patient had a tumour in the floor of the nasal cavity with invasion of the palate on biopsy but not imaging. Cervical lymphadenopathy was seen in two cases, and the nasopharynx was normal in all the four patients. CONCLUSION: Lymphoepitheliomas occur in the region of the palate, where they are also EBV related in southern Chinese. Compared with the keratinizing squamous cell carcinomas, patients with lymphoepitheliomas have a better prognosis and these tumours are not tobacco or alcohol related. They should not be misdiagnosed as metastatic nasopharyngeal carcinoma (NPC), particularly since the nasopharynx is invariably normal on imaging and adequate nasopharyngeal biopsy is negative for malignancy. PMID- 10362234 TI - Detection and follow-up of important extra-arterial lesions with helical CT angiography. AB - AIM: To determine the prevalence and significance of extra-arterial findings detected prospectively on helical computed tomographic angiography (CTA). SUBJECTS AND METHODS: The official reports of 802 consecutive CTAs performed over a 4.5-year period on 624 patients and the reports of all radiographic follow-up studies were reviewed for identification of important extra-arterial findings. Medical records and imaging studies of all patients with previously unknown extra arterial findings on CTA were reviewed to assess follow-up. In cases where follow up was not indicated in the medical record, referring physicians were contacted directly. RESULTS: Important, previously unknown, extra-arterial findings were detected on 35 CTAs (4.4% of all CTAs, 5.6% of all patients), with 33 of 35 detected prospectively. Six lesions were consistent with and/or proven to be malignant. Important non-tumoural lesions were discovered on nine CTAs. Of 13 lesions with imaging features that were suspicious for malignancy. Five of these lesions proved to be benign, but radiographic and/or clinical follow-up was not obtained or could not be documented in eight patients. CONCLUSION: With the increasing use of CTA as a replacement for conventional angiography, careful attention should be paid to the visualized extra-arterial structures. Extra vascular findings that are believed to be significant, may not be adequately followed-up by referring vascular specialists. PMID- 10362235 TI - Somatomotor fMRI in the pre-surgical evaluation of a case of focal epilepsy. AB - A child with intractable partial epilepsy who was found to have a focal lesion in the motor cortex underwent detailed pre-surgical and intraoperative investigations which enabled curative surgery without morbidity by minimizing a targeted resection. The pre-surgical assessment included mapping motor cortical function with functional magnetic resonance imaging (fMRI). This was subsequently correlated with the results of pre-surgical and intraoperative invasive corticography. PMID- 10362236 TI - MR imaging of thyroglossal duct cysts in adults. AB - AIM: To describe the magnetic resonance (MR) features of thyroglossal duct cysts (TDC) in adults. PATIENTS AND METHODS: Sixteen patients with TDC underwent MR imaging to obtain T1- and T2-weighted images and T2-weighted fat saturation images. In addition, contrast enhanced images were obtained in five patients. RESULTS: The signal intensity of TDC was of that of a simple cyst in seven (44%) patients, yielding high signal intensity on T2- and low signal intensity on T1 weighted images. In nine (56%), the signal intensity was either intermediate or high on T1-weighted images, the T2 signal intensity in these cases being high (7), intermediate (1) or low (1). Enhancement of the wall of the cyst was present in three of five (60%) patients. All 16 TDC were located at or just to one side of the midline and 12 were embedded in the strap muscles. All TDC were infrahyoid in location but 11 also extended superiorly to be directly related to the hyoid. At the hyoid the cystic component was immediately posterior (6) anterior (3) or anterior and posterior (2) to the bone. Intralaryngeal extension was present in eight (50%) patients. A suprahyoid tract was identified in three patients. The thyroid gland was in a normal location in all patients. CONCLUSION: Thyroglossal duct cysts are most commonly of high T1 signal intensity consistent with high protein content. The tract leading to the base of the tongue is infrequently seen, the diagnosis being determined by the intimate relationship to the hyoid and strap muscles. Intralaryngeal extension in adult patients with TDC is more frequent than reported previously. PMID- 10362237 TI - Two cases of seminal vesicle fistula. AB - Two cases of fistulation into the seminal vesicles are described. One related to Crohn's disease and the other following surgery for carcinoma of the rectum. Both cases were diagnosed by CT sinography. This technique is described and is recommended when attempting to demonstrate the internal communications of difficult perineal fistulae when standard techniques of fistulography fail. PMID- 10362238 TI - Does a joint ultrasound guided cytology clinic optimize the cytological evaluation of head and neck masses? AB - AIM: To compare the results of fine-needle aspiration (FNA) of head and neck masses performed in an ultrasound-guided cytology clinic (USGCC) staffed by a radiologist and pathologist to those obtained with specimens sent from other sources. METHODS: Comparison of broad-category FNA diagnoses (malignant, uncertain, benign or inadequate) with the patient's ultimate clinical or pathological outcome. Because FNA outcomes are semi-quantitative, accuracy of the procedure (the proportion of all tests resulting in a true- positive or negative fine-needle aspirate) is a better measure than sensitivity or specificity. Specimens (n = 292) from the first 2 years of the USGCC are compared with 600 specimens received from other sources over the previous 4 years. RESULTS: Accuracy was 23.4% better for specimens from the USGCC compared with those obtained by clinician guided aspiration (83.9%, 95% CI 79.7-88.1%, vs 60.5%, 95% CI 56.6-64.4%). There was an 84% reduction in inadequate specimens (from 21.5% to 3.4%). The proportion resulting in an uncertain result did not alter; 12.0% for USGCC and 11.9% for clinician-derived specimens. Improvement in accurate identification of salivary gland, lymph node, soft tissue and thyroid pathology was 27.0%, 21.2%, 18.3% and 15.8% respectively. CONCLUSIONS: The common practice of FNA performed by clinicians produces sub-optimal results in head and neck masses. A combined approach of ultrasound-guided fine-needle aspiration of head and neck masses, with immediate assessment of the material by a pathologist, is more accurate than with specimens obtained in other ways. If the results of FNA are to be incorporated into clinical decision making, the samples are best obtained using the USGCC model. PMID- 10362239 TI - Scrotal trauma: a cause of testicular atrophy. AB - Scrotal trauma is often mentioned as a cause of testicular atrophy yet there have been few studies documenting the effect of scrotal trauma on testicular size months or years following injury. We performed clinical and sonographic examinations in 10 patients who had suffered blunt scrotal trauma. A significant reduction in volume of the injured testis was observed in 5/10 cases at follow-up sonography. In two cases the affected testis was heterogeneous and colour flow Doppler examination showed reduced flow. In three cases the testis was homogeneous but reduced in volume and in the remaining five cases the affected testis appeared normal. We conclude that testicular atrophy is a sequel of scrotal trauma and occurred in 50% of patients in this study. PMID- 10362240 TI - With advances in medical imaging can the radiologist reliably diagnose Wilms' tumours? AB - AIM: To evaluate whether radiologists can accurately differentiate Wilms' tumours from other paediatric abdominal masses with renal involvement using modern imaging methods alone. METHODS: From February 1993 to June 1997, 23 patients presented to the Paediatric Oncology Service at The Royal Hospital for Sick Children, Edinburgh with an intra-abdominal mass which had renal involvement. Nine patients had Wilms' tumours, 12 had neuroblastomas, one patient had xanthogranulomatous pyelonephritis and there was a single case of a mesoblastic nephroma. In each case, two radiologists retrospectively reviewed the initial imaging examinations and independently reached a radiological diagnosis. RESULTS: The radiologists were concordant and reached the correct diagnosis in 20/23 cases (87%), unsure of the diagnosis in one case (4.3 %) and discordant in two cases (8.7 %). Radiologists should be aware that a mesoblastic nephroma can have identical imaging features to a Wilms' tumour. In most cases, ultrasound and a chest X-ray were sufficient to reach the correct diagnosis although computed tomography (CT) and magnetic resonance imaging were superior for demonstrating the relationship of the mass to the great vessels, retroperitoneum and spinal canal. Inferior vena cava invasion was strongly predictive of a Wilms' tumour. Displacement of the great vessels, extension of the mass across the mid-line, renal displacement and tumour calcification on CT were more suggestive of a neuroblastoma although these features were also seen in a significant number of patients with Wilms' tumours. Encasement of vessels by tumour, a paravertebral mass and spinal canal invasion were highly predictive of neuroblastoma. CONCLUSION: In this study, radiologists were accurate at diagnosing Wilms' tumours using modern imaging methods, however, care should be taken in children who are less than 1 year of age as a mesoblastic nephroma may have identical imaging characteristics. PMID- 10362241 TI - Large needle core biopsy--avoidance of biopsy table bombardment by the biopsy needle in stereotactic guided breast biopsy. AB - Table bombardment by automated biopsy needle is an occasional technical problem during stereotactic core biopsy of the breast. It is most commonly encountered in small breasts and when lesions are close to the biopsy table. A reference chart is devised for stereotactic core biopsy using an add-on erect stereotactic biopsy table and automated spring-loaded biopsy gun. The chart serves as a reference for pre-biopsy assessment of the feasibility of core biopsy in the vertical approach due to the constraints of lesion depth and breast thickness. It allows easy and quick reference for the required needle length input for the stereotactic system to prevent table bombardment. PMID- 10362242 TI - Solitary caecal ulcer syndrome, a benign condition which mimics the CT appearance of caecal carcinoma. PMID- 10362243 TI - Unilateral appearance of markedly dilated Virchow-Robin spaces. PMID- 10362244 TI - Conservative surgical removal of a submandibular duct calculus following interventional sialography. PMID- 10362245 TI - Retrorectal cystic hamartoma: diagnosis using endorectal ultrasound. PMID- 10362246 TI - Venous reflux in body computed tomography. PMID- 10362247 TI - Day case angiography of anticoagulated patients using the Angio-Seal haemostatic puncture closure device. PMID- 10362248 TI - Thymoliposarcoma: CT and pathologic findings. PMID- 10362249 TI - p21Waf1/Cip1/Sdi1 induces permanent growth arrest with markers of replicative senescence in human tumor cells lacking functional p53. AB - We have shown previously that wild type p53 can rapidly induce replicative senescence in EJ human bladder carcinoma cells lacking functional p53. A major effector of p53 functions is p21Waf1/Cip1/Sdi1, a potent cyclin-dependent kinase inhibitor. p21Waf1/Cip1/Sdi1 has been shown to be involved in both p53 dependent and independent control of cell proliferation, differentiation and death. To directly investigate the effects of p21Waf1/Cip1/Sdi1 in the p53 response observed in EJ tumor cells, we established p21Waf1/Cip1/Sdi1 inducible lines using the tetracycline-regulatable vector system. p21Waf1/Cip1/Sdi1 induction caused irreversible cell cycle arrest in both G1 and G2/M, and diminished Cdk2 kinase activity. In addition, p21Waf1/Cip1/Sdi1 induction led to morphological alterations characteristic of cells undergoing replicative senescence with morphological, biochemical and ultrastructural markers of the senescent phenotype. Furthermore, sustained p21Waf1/Cip1/Sdi1 induction sensitized EJ cells to apoptotic cell death induced by mitomycin C, a cross-linking DNA damaging agent. These findings support the function of p21Waf1/Cip1/Sdi1 as an inducer of replicative senescence and a major mediator of this phenomenon in response to p53. Moreover, our results imply that therapeutic intervention in human cancers might be aimed at sustained elevation of p21Waf1/Cip1/Sdi1 expression. PMID- 10362250 TI - Molecular mechanisms underlying interferon-alpha-induced G0/G1 arrest: CKI mediated regulation of G1 Cdk-complexes and activation of pocket proteins. AB - One prominent effect of IFNs is their cell growth-inhibitory activity. The mechanism behind this inhibition of proliferation is still not fully understood. In this study, the effect of IFN-alpha treatment on cell cycle progression has been analysed in three lymphoid cell lines, Daudi, U-266 and H9. Examination of the growth-arrested cell populations shows that Daudi cells accumulate in a G0 like state, whereas U-266 cells arrest later in G1. H9 cells are completely resistant to IFN-alpha's cell growth-inhibitory effects. The G0/G1-phase arrest is preceded by a rapid induction of the cyclin-dependent kinase inhibitors (CKIs), p21 and p15. In parallel, the activities of the G1 Cdks are significantly reduced. In addition to p21/p15 induction, IFN-alpha regulates the expression of another CKI, p27, presumably by a post-transcriptional mechanism. In the G1 Cdk complexes, there is first an increased binding of p21 and p15 to their respective kinases. At longer exposure times, when Cdk-bound p15 and p21 decline, p27 starts to accumulate. Furthermore, we found that IFN-alpha not only suppresses the phosphorylation of pRb, but also alters the phosphorylation and expression of the other pocket proteins p130 and p107. These data suggest that induction of p21/p15 is involved in the primary IFN-alpha response inhibiting G1 Cdk activity, whereas increased p27 expression is part of a second set of events which keep these Cdks in their inactive form. Moreover, elevated levels of p27 correlated with a dissociation of cyclin E/Cdk2-p130 or p107 complexes to yield cyclin E/Cdk2-p27 complexes. In resistant H9 cells, which possess a homozygous deletion of the p15/p16 genes and lack p21 protein expression, IFN-alpha causes no detectable changes in p27 expression and, furthermore, no effects are observed on either pocket proteins in this cell line. Taken together, these data suggest that the early decline in G1 Cdk activity, subsequent changes in phosphorylation of pocket proteins, and G1/G0 arrest following IFN-alpha treatment, is not primarily due to loss of the G1 kinase components, but result from the inhibitory action of CKIs on these complexes. PMID- 10362251 TI - Isolation and characterization of mammalian homologues of Caenorhabditis elegans lin-7: localization at cell-cell junctions. AB - In Caenorhabditis elegans, the vulval induction is mediated by the let-23 receptor tyrosine kinase (RTK)/ Ras signaling pathway. The precise localization of the let-23 RTK at the epithelial junctions is essential for the vulval induction, and requires three genes including lin-2, -7, and -10. The mammalian homologue of lin-2 has been identified as a protein interacting with a neuronal adhesion molecule, neurexin, and named CASK. CASK has recently been reported to interact with syndecans and an actin-binding protein, band 4.1, at epithelial and synaptic junctions, and to play central roles in the formation of cell-cell junctions. The product of C. elegans lin-7 directly interacts with let-23 RTK and localize it at epithelial junctions. Here, we report three rat homologues of lin 7 ubiquitously expressed in various tissues. These homologues are accumulated at the junctional complex region in cultured Madin-Darby canine kidney cells, and are also localized at the synaptic junctions in neurons. The mammalian homologues of lin-7 may be implicated in the formation of cell-cell junctions. PMID- 10362252 TI - Down-regulation of cyclin B1 gene transcription in terminally differentiated skeletal muscle cells is associated with loss of functional CCAAT-binding NF-Y complex. AB - The observation that cyclin B1 protein and mRNAs are down-regulated in terminally differentiated (TD) C2C12 cells, suggested us to investigate the transcriptional regulation of the cyclin B1 gene in these cells. Transfections of cyclin B1 promoter constructs indicate that two CCAAT boxes support cyclin B1 promoter activity in proliferating cells. EMSAs demonstrate that both CCAAT boxes are recognized by the trimeric NF-Y complex in proliferating but not in TD cells. Transfecting a dominant-negative mutant of NF-YA we provide evidence that NF-Y is required for maximal promoter activity. Addition of recombinant NF-YA to TD C2C12 nuclear extracts restores binding activity in vitro, thus indicating that the loss of NF-YA in TD cells is responsible for the lack of the NF-Y binding to the CCAAT boxes. Consistent with this, we found that the NF-YA protein is absent in TD C2C12 cells. In conclusion, our data demonstrate that NF-Y is required for cyclin B1 promoter activity. We also demonstrate that cyclin B1 expression is regulated at the transcriptional level in TD C2C12 cells and that the switch-off of cyclin B1 promoter activity in differentiated cells depends upon the loss of a functional NF-Y complex. In particular the loss of NF-YA protein is most likely responsible for its inactivation. PMID- 10362253 TI - The extracellular-signal-regulated protein kinases (Erks) are required for UV induced AP-1 activation in JB6 cells. AB - Mitogen activated protein (MAP) kinase belongs to a large family of serine/threonine protein kinases, including extracellular-signal-regulated protein kinases (Erks), P38 kinase and c-Jun N-terminal kinases (JNKs). Although previous work has shown that both Erks and JNKs are activated in cells in response to ultraviolet (UV) irradiation, most studies have focused only on the role of JNKs in UV-induced AP-1 activation. Hence, the role of Erks in UV-induced AP-1 activity is not well defined. We here have investigated this issue by using MAP kinase kinase (MEK1) inhibitor PD098059 and a dominant negative Erk2, as well as wild-type Erk2, in a JB6 cell model. PD098059 inhibited UVB- or UVC-induced AP 1 activity and phosphorylation of MEK1 and Erks, but not JNKs, in JB6 Cl 41 cells. Overexpression of wild-type Erk2 in Cl 30.7b cells that contain small amounts of Erks caused a 46.6- or 138.1-fold increase of AP-1 activity by UVB and UVC, respectively; introduction of a dominant negative Erk2 into Cl 41 cells significantly blocked the UV-induced Erks activation as well as the AP-1 activation. In contrast, overexpression of wild-type Erk2 in Cl 30.7b cells and dominant negative Erk2 in Cl 41 cells did not show a marked influence on the phosphorylation of JNKs. These results demonstrate that activation of Erks, in addition to the previously reported JNKs, is required for UV-induced AP-1 activation. PMID- 10362254 TI - In vivo analysis of the state of the human uPA enhancer following stimulation by TPA. AB - We have analysed in vivo the -2.0 kb enhancer of the human urokinase-type plasminogen activator (uPA) gene in HepG2 cells, in which gene expression can be induced by phorbol esters. The results reveal that, within the regulatory region, the enhancer, the silencer and the minimal promoter become hypersensitive to deoxyribonuclease I (DNase I) upon induction of transcription. The hypersensitivity of the enhancer can be reversed after removal of the inducer. In vivo footprinting analysis indicates that all the cis-acting elements of the enhancer, previously identified in vitro, are occupied in vivo upon 12-O tetradecanoyl-phorbol-13-acetate (TPA) stimulation of HepG2 cells. Micrococcal nuclease (MNase) cleavage of this region fails to reveal discrete nucleosomal boundaries in vivo in close proximity of the enhancer, either before or after stimulation. Furthermore, this region does not lose its nucleosomal configuration after TPA induction of transcription. An approximately 600 bp long region around the enhancer becomes more, but not fully, accessible to restriction endonucleases upon stimulation. A time-course experiment shows that this accessibility reaches a plateau after a 1 h TPA treatment suggesting the persistent presence of nucleosomes. These results indicate that TPA induces the binding of transcription factors to the uPA enhancer without chromatin remodelling of this region. PMID- 10362255 TI - Instabilities in phosphorylation-dephosphorylation cascades and cell cycle checkpoints. AB - The G2-M checkpoint in the cell cycle is identified with a set of phosphorylation dephosphorylation (PD) cycles involving Cdc25 and the maturation-promoting factor (MPF); these PD cycles are coupled in a way that generates an instability. This instability arises out of a transcritical bifurcation which could be exploited by the G2 DNA damage checkpoint pathway in order to arrest or delay entry into mitosis. The coupling between PD cycles involving Wee1 and MPF does not lead to an instability and therefore Wee1 may not be a crucial target of the checkpoint pathway. A set of PD cycles exhibiting transcritical bifurcation also possesses the integrative ability of a checkpoint for 'checking' that prerequisites are satisfied prior to the next cell cycle event. Such a set of coupled PD cycles is suggested to be a core mechanism of cell cycle checkpoints. PMID- 10362256 TI - Molecular analysis of an unstable genomic region at chromosome band 11q23 reveals a disruption of the gene encoding the alpha2 subunit of platelet-activating factor acetylhydrolase (Pafah1a2) in human lymphoma. AB - A region of 150 kb has been analysed around a previously isolated, lymphoma associated, translocation breakpoint located at chromosome band 11q23. This balanced and reciprocal translocation, t(11;14)(q32;q23), has been shown to result in the fusion between chromosome 11 specific sequence and the switch gamma4 region of the IGH locus. The LPC gene, encoding a novel proprotein convertase belonging to the furin family, has been identified in this region. In order to characterize further the region surrounding the translocation, we have determined the detailed structure of LPC. Here we show that LPC consists of at least 16 exons covering 25 kb, and that there is a partial duplication, involving mobile genetic elements and containing LPC exons 13-17 in a tail-tail configuration at 65 kb downstream. Since the chromosomal breakpoint lay between these two structures, the intervening region was further analysed and shown to contain at least two unrelated genes. The previously known SM22 gene was localized close to the 3' tail of LPC. Furthermore, we identified the gene encoding the alpha2 subunit of platelet-activating factor acetylhydrolase (Pafah1a2) at the chromosomal breakpoint. The position of another previously identified breakpoint was also located to within the first intron of this gene. Altogether, our results give evidence of a genomic instability of this area of 11q23 and show that Pafah1a2 and not LPC is the gene disrupted by the translocation, suggesting that deregulated Pafah1a2 may have a role in lymphomagenesis. PMID- 10362257 TI - The proapoptotic effect of hepatitis B virus HBx protein correlates with its transactivation activity in stably transfected cell lines. AB - The role of hepatitis B virus HBx protein in the carcinogenesis associated with chronic viral infection remains ill-defined. Indeed, pleiotropic effects have been ascribed to HBx: in addition to its well-documented ability to indirectly stimulate transcription, the protein has been reported to affect cell growth, signal transduction, DNA repair and apoptosis. In this work, we generated Chang (CCL-13)-derived cell lines constitutively expressing wild type or mutant HBx, as a model of HBx-host cell interaction closer to the chronic infection setting, than the classically used transient expression systems. We document the potentiation by HBx of the apoptotic cell death pathway in the recipient cells. This effect is unlikely to rely on p53 activity since the protein is functionally inactivated in CCL-13. In addition, antioxidants and cyclosporin A failed to reduce the apoptotic response back to the normal level, suggesting that production of reactive oxygen species and calcineurin activation are not directly involved in the proapoptotic effect of HBx. In contrast, our data show that transactivation and stimulation of apoptosis are tightly linked HBx activities. Finally, expression of transactivation-active protein did not result in detectable change in the pattern of MAP kinases phosphorylation nor did it affect the ability of the host cell to repair in vitro irradiated plasmid DNA. PMID- 10362258 TI - Cooperation of Sp1 and p300 in the induction of the CDK inhibitor p21WAF1/CIP1 during NGF-mediated neuronal differentiation. AB - Addition of nerve growth factor (NGF) to PC12 cells promotes neuronal differentiation while inhibiting cell proliferation. In order to understand how NGF exerts its antimitogenic effect during differentiation, we have studied the mechanism by which this factor activates the promoter of the CDK inhibitor p21W4F1/CIP1. The minimal region of the p21 promoter required for the NGF induction was mapped to a contiguous stretch of 10 bp located 83 bases upstream of the transcription initiation site. This GC-rich region was shown to interact specifically with the transcription factor Sp1 and the related protein Sp3, in either exponentially-growing or NGF-treated PC12 cells. The addition of NGF resulted in an accumulation of the transcriptional co-activator p300 in complexes associated with the NGF-responsive region. Transcriptional activity of Sp1, Sp3 and p300 was specifically induced by NGF in a Gal4-fusion assay, indicating that induction of p21 during neuronal differentiation may involve regulation of the activity of these factors by NGF. Furthermore, p300 was able to act as a co activator for Sp1-mediated transcriptional activation in PC12 cells, suggesting that p300 and Sp1 may cooperate in activating p21 transcription during the withdrawal of neuronal precursors from the cell cycle. This hypothesis is supported by experiments showing that p300 and Sp1 form complexes in PC12 cells. PMID- 10362259 TI - The metalloproteinase matrilysin is a target of beta-catenin transactivation in intestinal tumors. AB - Matrilysin is a matrix metalloproteinase expressed in the tumor cells of greater than 80% of intestinal adenomas. The majority of these intestinal tumors are associated with the accumulation of beta-catenin, a component of the cadherin adhesion complex and, through its association with the T Cell Factor (Tcf) DNA binding proteins, a regulator in the Wnt signal transduction pathway. In murine intestinal tumors, matrilysin transcripts show striking overlap with the accumulation of beta-catenin protein. The matrilysin promoter is upregulated as much as 12-fold by beta-catenin in colon tumor cell lines in a manner inversely proportional to the endogenous levels of beta-catenin/Tcf complex and is dependent upon a single optimal Tcf-4 recognition site. Coexpression of the E cadherin cytoplasmic domain blocked this induction and reduced basal promoter activity in every colon cancer cell line tested. Inactivation of the Tcf binding site increased promoter activity and overexpression of the Tcf factor, LEF-1, significantly downregulated matrilysin promoter activity, suggesting that beta catenin transactivates the matrilysin promoter by virtue of its ability to abrogate Tcf-mediated repression. Because genetic ablation of matrilysin decreases tumor formation in multiple intestinal neoplasia (Min) mice, we propose that regulation of matrilysin production by beta-catenin accumulation is a contributing factor to intestinal tumorigenesis. PMID- 10362260 TI - Association with Cdc2 and inhibition of Cdc2/Cyclin B1 kinase activity by the p53 regulated protein Gadd45. AB - Recently Gadd45, a p53-regulated stress protein, has been implicated in the activation of a G2/M checkpoint after damage by UV radiation and alkylating agents. While inhibitory phosphorylation of Cdc2 and suppression of cyclin B1 levels are known to be involved in G2 delays after genotoxic stress, Gadd45 has now been found to directly inhibit the activity of Cdc2/Cyclin B1 complex, while it had no appreciable effect on Cdk2/ Cyclin E activity even at very high levels of Gadd45. In contrast, p21CiP1/Waf1 is an universal cdk/cyclin inhibitor and inhibited both of the cyclin complexes tested here. Gadd45 was also able to physically interact with Cdc2, but not Cyclin B1. Addition of Gadd45 to immunoprecipitated Cdc2/Cyclin B1 in vitro led to a dissociation of this complex, and thus may represent a new checkpoint mechanism whereby Cdc2/Cyclin B1 can be inhibited. With the use of an antisense approach, reduced Gadd45 expression attenuated the suppression of Cdc2/Cyclin B1 activity in UV-irradiated human cells. Taken together, these results implicate Gadd45 in the control of G2/M cell cycle progression after certain stresses. PMID- 10362261 TI - Accentuated apoptosis in normally developing p53 knockout mouse embryos following genotoxic stress. AB - In order to identify the alternative pathways which may substitute for the p53 function during embryogenesis, we have focused our studies on p53 -/- normally developing mouse embryos that survived a genotoxic stress. We assumed that under these conditions p53-independent pathways, which physiologically control genomic stability, are enhanced. We found that while p53 +/+ mouse embryos elicited, as expected, a p53-dependent apoptosis, p53-/- normally developing mice exhibited an accentuated p53-independent apoptotic response. The p53-dependent apoptosis detected in p53+/+ embryos, was an immediate reaction mostly detected in the brain, whereas the p53-independent apoptosis was a delayed reaction with a prominent pattern observed in epithelial cells of most organs in the p53 deficient mice only. These results suggest that in the absence of p53-dependent apoptosis, which is a fast response to damaged DNA, p53-independent apoptotic pathways, with slower kinetics, are turned on to secure genome stability. PMID- 10362262 TI - The proto-oncogene c-Cbl is a negative regulator of DNA synthesis initiated by both receptor and cytoplasmic tyrosine kinases. AB - In C. elegans, genetic and biochemical data indicate that the Cbl homolog Sli-1 attenuates Let-23 (EGFR) signaling. To investigate whether c-Cbhl might have a role in mammalian growth factor-mediated mitogenic signaling, we microinjected NIH3T3 mouse fibroblasts with expression plasmids encoding wt and G306ECbl (a 'loss of function' mutant identified in C. elegans). We observed inhibition of PDGF BB- and EGF-induced DNA synthesis by wt Cbl but not the mutant. Microinjection of two different affinity purified polyclonal antisera against Cbl boosted a suboptimal PDGF-stimulated mitogenic response. The inhibition of both PDGF BB- and EGF-induced DNA synthesis by wt Cbl was reversed by co-expression with Myc but not with Fos. DNA synthesis initiated by constitutively activated Src was also blocked by Cbl expression, but curiously by the G306E mutant as well. These data are all consistent with the proposition that Cbl negatively affects mitogenic signaling in mammalian fibroblasts. PMID- 10362263 TI - Combined chelation therapy in reducing tissue lead concentrations in suckling rats. AB - The very young are more prone to lead poisoning than adults, and the treatment with chelating agents, either as monotherapy or combined treatment, is still a matter of dispute. The purpose of this work was to evaluate the efficiency of three chelating agents administered either as monotherapies or as combined treatments in sucklings. Lead acetate (5 mg Pb kg(-1) i.p.) was administered to the 7-day-old rat pups in eight litters on experimental day 1 and chelating agents on experimental days 2 and 3. Pups were divided into six groups: (1) untreated control; (2) EDTA (calcium disodium ethylendiaminetetraacetate, 0.3 mmol kg(-1) i.p. at 4 p.m.); (3) meso-DMSA (meso-2,3-dimeracaptosuccinic acid, 0.5 mmol kg(-1) p.o. at 10 a.m.); (4) rac-DMSA (racemic-2,3-dimeracaptosuccinic acid, 0.5 mmol kg(-1) p.o. at 10 a.m.); (5) EDTA+meso-DMSA; and (6) EDTA+rac DMSA. Rats were killed on experimental day 5. Tissue element concentrations were analyzed by atomic absorption spectrometry. Treatment with EDTA did not affect tissue Pb, but it reduced Zn in the carcass and liver. Meso-DMSA reduced Pb in the kidneys and brain, and it did not affect organ essential elements. Rac-DMSA most efficiently reduced Pb concentrations in the carcass, kidneys and brain, but it also reduced Zn and Cu in the liver and Zn in the kidneys. Combined treatments with EDTA never improved the efficiency of either DMSA isoform in decreasing tissue Pb but they did reduce tissue Zn concentrations. All treatments caused the same decrease in the carcass Ca concentrations. The results do not support combined treatment in this age group, which is especially sensitive to trace element deficiencies, and suggest that meso-DMSA might be the treatment of choice in acute lead poisoning in infants. PMID- 10362264 TI - Cytokine endpoints for the local lymph node assay: consideration of interferon gamma and interleukin 12. AB - The murine local lymph node assay (LLNA) is a method for the prospective identification of contact allergens. Skin sensitization potential is assessed as a function of induced proliferative responses in lymph nodes draining the site of topical exposure measured in situ by incorporation of radiolabelled thymidine ([3H]thymidine). The results of previous investigations have demonstrated that the analysis of [3H]thymidine incorporation represents a robust and reliable endpoint for the LLNA for the assessment of skin sensitizing activity for strong and moderate allergens and, in addition, many weaker sensitizers. The aim of the current experiments was to explore the utility of the production of the cytokines interferon-gamma (IFN-gamma) and interleukin 12 (IL-12) by draining lymph node cells (LNC) as alternative readouts for the LLNA. Animals were exposed to a range of skin sensitizers at two application concentrations. The first of these was chosen on the basis of results from previous investigations to stimulate a strong proliferative response (tenfold or greater increase in proliferation compared with concurrent vehicle controls). The second concentration of test material in each case was the amount of chemical estimated to be necessary mathematically for elicitation of a stimulation index of 3 (EC3 value); the induction of a threefold or greater increase in proliferation is the current criterion for a positive response in the LLNA. In addition, analyses were conducted with para-aminobenzoic acid (PABA), a non-sensitizing chemical shown previously not to induce LLNA responses. Secretion of IFN-gamma and the p40 subunit of IL-12 by draining LNC was measured by cytokine-specific enzyme-linked immunosorbent assay. In parallel experiments, LNC activity was assessed as a function of [3H]thymidine incorporation in situ. All the chemical allergens tested provoked robust proliferative responses, with the stimulation indices recorded at both test concentrations reflecting only small changes in activity compared with previously recorded data. Exposure to vehicle (4:1 acetone:olive oil, AOO) alone resulted in detectable, although variable, expression of both IFN-gamma and IL-12. Treatment with chemical allergen in each case caused a marked increase in IFN-gamma secretion, with particularly vigorous production of cytokine being stimulated following exposure to oxazolone or hexyl cinnamic aldehyde. In contrast, application of chemical allergens was not generally associated with elevated IL 12 p40 secretion. Exposure of mice to PABA did not result in increased IFN-gamma or IL-12 production compared with vehicle-treated controls. In general, however, cytokine secretion did not correlate closely with the induction of LNC proliferation. These data indicate that expression by allergen-activated LNC of IFN-gamma or IL-12 does not provide a reliable or sufficiently sensitive endpoint for the LLNA compared with [3H]thymidine incorporation in situ. PMID- 10362265 TI - Pharmacokinetics of antipyrine, warfarin and paracetamol in the brushtail possum. AB - The plasma pharmacokinetics of antipyrine, warfarin and paracetamol have been studied in the Australian brushtail possum (Trichosurus vulpecula). The plasma elimination half-lives (t1/2) were 1.2 h for antipyrine, 11.9 h for warfarin and 5.2-12.9 h for paracetamol. Our data indicate that the clearance of these three xenobiotics in the possum is similar to that reported in eutherian mammals. There was no dose-dependent increase in paracetamol plasma t1/2 over the dose range 100 1000 mg kg(-1), indicating a lack of capacity saturation. This observation may in part explain the unusual resistance of the possum to the hepatotoxic effect of high doses of paracetamol. PMID- 10362266 TI - Comparison of cytotoxicity of various surfactants tested on normal human fibroblast cultures using the neutral red test, MTT assay and LDH release. AB - We used the neutral red test, MTT assay and lactate dehydrogenase (LDH) release to compare the potential cytotoxicity of six surfactants belonging to different classes--three non-ionic surfactants (Triton x100, octylphenoxypolyethoxy alcohol, from Orion; Tween 60, polyoxyethylene (20) sorbitan monostearate, from ICI Speciality Chemicals; Tween 80, polyoxyethylene (20) sorbitan monolaurate, from Labosi), two anionic surfactants (Texapon K1298, sodium lauryl sulphate, from Henkel; Texapon N40, sodium laurylether sulphate, from Henkel) and one cationic surfactant (benzethonium chloride, from Siber Hegner)--on human fibroblast cultures. According to the LC50 (microg ml(-1)), the tested surfactants can be classified in the following order of increasing cytotoxicity: Tween 80 < Texapon N40 < Tween 60 < Texapon K1298 < Triton x100 < benzethonium chloride. PMID- 10362267 TI - Neurodevelopmental evaluation of 9-month-old infants exposed to low levels of lead in utero: involvement of monoamine neurotransmitters. AB - The objective of this work is to investigate the neurotoxicty of low-level lead exposure in utero on infants and the possible involvement of dopaminergic and serotonergic neurotransmitters. The correlation analysis for cord blood lead level, the concentrations of dopamine metabolite homovanillic acid (HVA) and serotonin metabolite 5-hydroxyindoleacetic acid (5-HIAA) in cord plasma and the neurodevelopmental scales of infants were conducted on 244 9-month-old children. Both score of sociability subscale and 5-HIAA concentration were correlated with cord blood lead level. The sociability score was negatively correlated with the concentration of HVA, whereas both the coordination score and the global score were negatively correlated with the concentration of 5-HIAA. With partial correlation analysis, after taking HVA into account, the significant negative correlation between the sociability score and the cord blood lead level that existed in the linear correlation analysis disappeared, and the score of global scale correlated negatively with lead level in cord blood. When taking 5-HIAA into account, the scores of all the neurodevelopmental subscales except the language subscale were significantly negatively correlated with lead level in cord blood. The results indicated that low-level lead exposure in utero could produce a neurotoxic effect on the developing serotonergic system in infants. The neurotoxicity of low-level lead exposure in utero may affect the sociability of infants. Serotonergic activity was shown to have a potential effect on neurodevelopmental assessment. It may interfere with the association between low level lead exposure in utero and other neurodevelopmental performances of 9-month old children. PMID- 10362268 TI - In vitro and in vivo comparison of sulfur donors as antidotes to acute cyanide intoxication. AB - Antidotes for cyanide (CN) intoxication include the use of sulfane sulfur donors (SSDs), such as thiosulfate, which increase the conversion of CN to thiocyanate by the enzyme rhodanese. To develop pretreatments that might be useful against CN, SSDs with greater lipophilicity than thiosulfate were synthesized and assessed. The ability of SSDs to protect mice against 2LD50 of sodium cyanide (NaCN) administered either 15 or 60 min following administration of an SSD was assessed. To study the mechanism of action of the SSD, the candidate compounds were examined in vitro for their effect on rhodanese and 3-mercaptopyruvate sulfurtransferase (MST) activity under increasing SSD concentrations. Tests were conducted on nine candidate SSDs: ICD1021 (3-hydroxypyridin-2-yl N-[(N-methyl-3 aminopropyl)]-2-aminoethyl disulfide dihydrochloride), ICD1022, (3-hydroxypyridin 2-yl N-[(N-methyl-3-aminopropyl)]-2-aminoethyl disulfide trihydrochloride), ICD1584 (diethyl tetrasulfide), ICD1585 (diallyl tetrasulfide), ICD1587 (diisopropyl tetrasulfide); ICD1738 (N-(3-aminopropyl)-2-aminoethyl 2-oxopropyl disulfide dihydrochloride), ICD1816 (3,3'-tetrathiobis-N-acctyl-L-alanine), ICD2214 (2-aminoethyl 4-methoxyphenyl disulfide hydrochloride) and ICD2467 (bis(4 methoxyphenyl) disulfide). These tests demonstrated that altering the chemical substituent of the longer chain sulfide modified the ability of the candidate SSD to protect against CN toxicity. At least two of the SSDs at selected doses provided 100% protection against 2LD50 of NaCN, normally an LD99. All compounds were evaluated using locomotor activity as a measure of potential adverse behavioral effects. Positive hypoactivity relationships were found with several disulfides but none was found with ICD1584, a tetrasulfide. Separate studies suggest that the chemical reaction of potassium cyanide (KCN) and cystine forms the toxic metabolite 2-iminothiazolidine-4-carboxylic acid. An alternative detoxification pathway, one not primarily involving the sulfur transferases. may be important in pretreatment for CN intoxication. Although studies to elucidate the precise mechanisms are needed. it is clear that these newly synthesized compounds provide a new rationale for anti-CN drugs, with fewer side-effects than the methemoglobin formers. PMID- 10362269 TI - Assessment of molybdenum toxicity in humans. AB - In an attempt to define a tolerable daily intake (TDI) for molybdenum based on a toxicological risk analysis approach, a large literature survey was conducted. In man, absorption of molybdenum after oral intake is in the range of 28-77% and urinary excretion is 17-80% of the total dose. A low order of toxicity of molybdenum compounds has been observed in humans. However, with the available data, it is not possible to calculate any dose-response or dose-effect relationships. Because molybdenum toxicity is associated with copper intake or depleted copper stores in the body, humans who have an inadequate intake of dietary copper or some dysfunction in their copper metabolism that makes them copper-deficient could be at greater risk of molybdenum toxicity. In the absence of relevant human studies, animal studies were evaluated for the derivation of the TDI. Effects of Mo on reproduction and foetal development were found to be critical effects observed in rats and mice. A dose-response relationship was observed in a study by Fungwe et al., with a 'no observed adverse effect' level (NOAEL) and a 'lowest observed adverse effect' level (LOAEL) of 0.9 and 1.6 mg Mo kg(-1) day(-1), respectively. Applying uncertainty factors of 10 for intraspecies and 10 for interspecies differences to the NOAEL, a TDI of 0.009 mg Mo kg(-1) day(-1) was calculated. The TDI is given a medium confidence rating. This TDI is more than double the upper limit of adequate intake for adolescents and adults that was derived from the Mo content of the average diet in the USA. PMID- 10362270 TI - Metabolism-dependent hepatotoxicity of methimazole in mice depleted of glutathione. AB - Methimazole (MMI) (>0.1 mmol kg(-1), p.o.) given in combination with DL buthionine sulphoximine (BSO) (3 mmol kg(-1), i.p., 1 h before MMI administration), an inhibitor of glutathione (GSH) synthesis, caused liver injury in mice. The injury was characterized by centrilobular necrosis of hepatocytes and an increase in serum alanine transaminase (ALT) activity. Methionazole (2 mmol kg(-1)) alone resulted in only a marginal increase in serum ALT activity, but produced no histopathological changes in the liver. Pretreatment with hepatic cytochrome P-450 monooxygenase inhibitors--cobalt chloride, isosafrole, methoxsalen, metyrapone and piperonyl butoxide-prevented or tended to suppress the hepatotoxicity induced by MMI in combination with BSO. Treatment with N,N dimethylaniline and ethyl methyl sulphide, competitive substrates of flavin containing monooxygenases (FMO), also resulted in remarkable suppression of the hepatotoxicity caused by MMI in combination with BSO. These results suggest that MMI is activated by reactions mediated by both cytochrome P-450 monooxygenases and FMO, and that the inadequate rates of detoxification of the resulting metabolite are responsible for the hepatotoxicity in GSH-depleted mice. PMID- 10362271 TI - Effect of aluminosilicates and bentonite on aflatoxin-induced developmental toxicity in rat. AB - Numerous studies have established that aflatoxin is a potent developmental toxin in animals. Previous research has demonstrated that a phyllosilicate clay, hydrated sodium calcium aluminosilicate (HSCAS or Novasil), tightly binds and immobilizes aflatoxins in the gastrointestinal tract of animals and markedly reduces the bioavailability and toxicity of aflatoxin. Our objective in this study was to utilize the pregnant rat as an in vivo model to compare the potential of HSCAS and bentonite to prevent the developmental toxicity of aflatoxin. Aluminosilicates (HSCAS) and bentonite were added to the diet at a level of 0.5% (w/w) and fed to the pregnant rat throughout pregnancy (i.e. days 0 20). Test animals were fed an aflatoxin-contaminated diet (2.5 mg kg(-1) diet) with or without sorbents during gestation days 6-15. Evaluations of toxicity were performed on day 20. These included maternal (mortality, body weights, feed intake and litter weights), developmental (embryonic resorptions and fetal body weights) and biochemical (ALT, AST and AP) evaluations. Sorbents alone were not toxic and aflatoxin alone resulted in significant maternal and developmental toxicity. Animals treated with phyllosilicate (plus aflatoxin) were comparable to controls following evaluations for resorptions, live fetuses and fetal body weights, as well as biochemical parameters. While bentonite plus aflatoxin resulted in significant reduction in fetal body weight, none of the fetuses from HSCAS or bentonite plus aflatoxin-treated groups had any gross, internal soft tissue or major skeletal malformations. PMID- 10362272 TI - Subchronic feeding study of antimony trioxide in rats. AB - Diets containing antimony trioxide were fed to male and female Wistar rats of the Alpk:APSD strain over 90 days. Dose levels were 0 (control), 1000, 5000 and 20,000 ppm (equivalent to mean daily doses of 84, 421 and 1686 mg kg(-1) in males and 97, 494 and 1879 mg kg(-1) in females). There was no effect of compound on growth or growth rate, food consumption or clinical signs. Minor changes in haematology and urine biochemistry were considered incidental to treatment. Small reductions in plasma alkaline phosphatase activity and increases in aspartate aminotransferase activity at the high dose, together with a small (ca. 10%) increase in liver weight, could be indicative of a minor effect on the liver, but in the absence of any histological effects these changes are also considered incidental to treatment. This study confirms the inert nature of antimony trioxide. PMID- 10362273 TI - Effects of in vitro cadmium exposure on ovarian steroidogenesis in rats. AB - The purpose of this study was to evaluate the direct effect(s) of in vitro cadmium (Cd) exposure on steroidogenesis in rat ovaries during different reproductive states. Sprague-Dawley rats were killed on the day of proestrus, or on gestation day 6 or 16. Ovaries were removed, placed in medium and minced. Culture from each ovary was incubated with Cd2+ ions in concentrations of 0, 100, 500, 1000, 1500, or 2000 microM. One-hour whole-ovary production of progesterone (P4), testosterone and estradiol (E2) in culture medium was evaluated in the absence and presence of human chorionic gonadotropin (hCG) or hCG plus pregnenolone by specific radioimmunoassay. Under in vitro Cd exposure the most affected were productions of P4 and testosterone in proestrus rats and less in pregnant dams, whereas E2 was not affected at all. Cadmium appears to interfere with the ovarian steroidogenic pathway in rats at more than one site. PMID- 10362274 TI - Biophysical shunt theory for neuropsychopathology: biphasal homeostatic dysregulation. AB - OBJECTIVE: We challenge Freud's psychodynamic theory using a systematic modus operandi which has been outlined in detail in a succession of articles. Here, we deal with Freud's first assumption of human psychological primacy in forming goal directed behavior. According to our theory, biphasal homeostatic dysregulation is the underlying mechanism of clinical phenomenology. MODEL: Evolutionary neurobiology has provided humans with a precise technical solution for optimal organismic survival. Humans are armed with an accurate negative feedback mechanism that operates within the alternating upper and lower thresholds of biphasal homeostatic maintenance and is coupled with a basal indicator of individual sensation of the degree of the given organismic well-being in any unit of time. This originates the organismic pleasure principle (OPP). The latter is achieved by a straightforward quantal injection of endorphins according to one of eight possible body operational regimens. Thanks to the essential duality of the dynamic interactions, stipulated by the complex harmonics of term-dependent and event-dependent adaptation when one or more of the essential elements for homeostasis goes above or below its predetermined threshold, certain branches of the organismic defense system (ODS) are 'turned on' in the second phase of homeostasis. The individual then adapts behavioral modifications directed toward a long-lasting search for the optimal resources needed for normal survival. This evolutionary biphasal homeostatic design has an intrinsic, methodical expression that confirms changes and correctly informs the individual about them, further imposing behavioral modifications, when necessary. In cases of a homeostatic derangement, the OPP is replaced by an erratic inclusion of pain, tension or depression, all components of the alarm system of the ODS, which may lead to disordered behavioral patterns. CONCLUSIONS: The underlying biological mechanism of goal-directed assignments for biphasal homeostatic maintenance is described. The intrinsic rules and regulations that guide both normal and abnormal survival may be clinically manifest. Normal survival behavior is necessary to regain organismic homeostasis. PMID- 10362275 TI - Biophysical shunt theory for neuropsychopathology: pentaphasic lipid-induced model for schizophrenia. AB - We propose a five-phase model for schizophrenia: (A) Imperfect diet-induced fatty acid combinations are mobilized from reservoirs during stress exposure into the bloodstream; (B) Increased brain-blood barrier permeability, with a further penetration of fat into the brain matter; (C) Rearrangement in the regional neuronal membrane fatty acids with a further physical compression of the adjacent membrane protein structures causing conformational alterations; (D) Cytokines generate further physical changes, impairing single- or multi-dispersed ion channels placed on the same neuron; (E) Ion channels are blocked, with a subsequent biophysical ion shunt-to-ion flow propagation, which results in neuronal misfiring and the appearance of schizophrenic signs and symptoms. PMID- 10362276 TI - Biophysical shunt theory for neuropsychopathology: neuroimmunological causality for the suicide condition. AB - This article describes a working hypothesis of the nature of the 'suicide condition' (SC). The authors contend that the SC emerges as a specialized result of two-phase impairment in the neuroimmunological 'inherited schematic representation' (ISR) involving: (a) the formation of a 'microcellular suicide' phenomenon; and (b) the establishment of a 'macro-organismic suicide' program. Our hypothesis, unlike earlier ones, is based on scientific evidence spanning diverse clinical diagnostic areas indicating that the SC is induced by a local histoincompatibility across distinct tissue structures and/or a remodeling of one or more neurimmunological ISR designs due to biophysical ion shunt bypass and neglect, whereas a normal neuroimmunological ISR complex produces an all embracing organismic histocompatible tissue-syntonic-to-ego-syntonic expression. The presence of abnormal 'microcellular suicide' assemblies leads to cell syntonic-to-cell dystonic transformation and initiates the first modification phase by contaminating certain neurimmunological ISR programs which in turn trigger the onset of partial ego syntonic-to-ego-dystonic conversion. This is translated by the self-conscious experience as partial self-to-alien tissue translocation. These formations accumulate at a rate, arriving at the second phase in SC establishment when they reach a magnitude resolution that surpasses the organismic suicide threshold level and increase the amount of ego-syntonic-to ego-dystonic inclusion. This is then translated by the self-conscious experience as a 'foreseeable and inescapable death' that is based on severe self-to-alien multiorgan substitution. The latter overcomes the rules and regulations prescribed by impulse-induced inner events, regardless of outer psychosocial events, and leads to an irrevocable drive to suicide. PMID- 10362277 TI - The role of brain insulin in the neurophysiology of serious mental disorders: review. AB - The purpose of this review is to indicate the role insulin plays in normal brain neurophysiology, together with the role insulin may play in the regulation of regional cerebral blood flow (rCBF). The relationship between sustained elevation of the inflammatory cytokines and brain insulin dysregulation, with respect to the serious mental disorders, is also discussed. It has been observed that, as the inflammatory cytokines increase, they exert a synergistic influence on insulin and somatostatin, by initially increasing and then decreasing insulin secretion. In the brain, increased levels of insulin result in increased glucose utilization and over-stimulation of the autonomic nervous system (ANS), while the inhibition of insulin secretion results in decreased glucose utilization and dysregulation of the hypothalamo-pituitary-adrenal (HPA) axis. It will further be argued that these alterations in brain insulin influence rCBF in the serious mental disorders such as schizophrenia and the affective disorders. It is hypothesized that insulin regulates rCBF either directly, or indirectly via GLUT4 in the hypothalamus now considered the glucose-sensing, insulin-sensing mechanism of the brain and the body. Thus, we shall propose that insulin plays an important role in normal neurophysiology and that sustained elevation of the inflammatory cytokines dysregulates insulin secretion, rCBF, ANS and the HPA-axis in serious mental disorders. PMID- 10362279 TI - Mouth bacteria as the cause of Paget's disease of bone. AB - The many viruses associated with Pagetic osteoclasts could be opportunistic rather than causative. Some mouth bacteria can lyse bone. One (Actinobacillus actinomycetemcomitans) can grow and even multiply inside human cell lines in culture, producing osteolytic materials -- one 62 kDa protein having a potency in the picomolar range. A small focus of this, or of one of the other periodontitis causing bacteria, in a bone might gradually spread its influence to activate osteoclasts -- the first stage in Paget's disease. The focus in each bone might be small and easily overlooked, as other intracellular bacteria have been in the past. PMID- 10362280 TI - From terra firma to terra plana--danger is shaking the foundations: deconstructing the 'immune system'. AB - The paradigm of an immune system presumes that a system arose specifically to combat infection -- hence its name. This paradigm gained credibility with the discovery of antibodies and anamnestic immunity, even though these are relatively late arrivals in evolution. Another presumption has been that thymus-dependent T cells are responsible for discriminating self from non-self. Subsequent opinion has crystallized around these presumptions. This paradigm is flawed. Transforming it into a morphostatic system resolves the problems. There is, arguably, no such thing as an immune system. PMID- 10362278 TI - Chromosomal fragility may be indicative of altered higher-order DNA organization as the underlying genetic diathesis in complex neurobehavioural disorders. AB - Preliminary observations concerning increased chromosomal fragility in association with certain behavioural disorders in humans allow an opportunity to suggest a cohesive theory regarding the possible importance of higher-order DNA modification in the coordination of gene function in brain evolution and during development. Visible or submicroscopic acentric chromosomal fragments are formed as an accompaniment to chromosomal breakage and are associated with sequence amplification. During genomic reintegration of such extra chromosomally amplified repeat sequence elements, functional consequences could include unequal crossing over with gain-of-function, and/or deletion with loss-of-function. This process could result in regulatory changes in gene function in association with normal coding regions, since fragile sites appear to be located at or near upstream DNase-I hypersensitive areas. Earlier research on chromosomal breakage in relation to transposon behaviour in maize has set a precedent by which many elements in a network could be coordinately controlled, a principle which may allow transcriptional control over multiple areas in the genome simultaneously. The hypothesis proposed in this paper implies that a small number of fundamental higher order changes may be responsible for influencing a wide range of genetic alterations leading to complex phenotypes, sometimes segregating as distinct entities within pedigrees, or alternatively, and perhaps more commonly, presenting with several overlapping phenotypes in some other families. Current emphasis on the investigation of only pure multiplex families in psychiatric genetics may assist with identification of a number of discrete behaviour modifying genes, but may not be sufficient for an understanding of the broad underlying genetic diathesis in these, and perhaps other 'multifactorial type' disorders. Validation of a role for altered fragile site expression and the molecular consequences thereof as proposed in this paper may offer additional avenues for gene therapy based either on preferential integration of exogenous DNA at fragile sites, or utilizing the acentric fragments formed during chromosome breakage to modify sequence amplification extrachromosomally. PMID- 10362281 TI - Azelaic acid: potential as a general antitumoural agent. AB - Azelaic acid is a naturally occurring straight-chained 9-carbon atom dicarboxylic acid which is non-toxic, non-teratogenic, and non-mutagenic. Its antiproliferative and cytotoxic effect on a variety of tumoural cell lines in culture, due to inhibition of mitochondrial oxidoreductases of the respiratory chain and of enzymes concerned with DNA synthesis is well established; normal cells are unaffected at similar dosages and times of exposure. Human melanoma cells xenotransplanted onto athymic nude mice are significantly affected by administration of azelaic acid. Clinically, in humans, it has already been shown to cause regression of melanoma in situ and primary invasive malignant melanoma. These results rank azelaic acid as a potential general antitumoural agent. It can be administered topically, focally, orally, intravenously, intra-arterially, and intralymphatically, all without local or general ill-effects, and is metabolized without harmful side-products. Simultaneous administration by different routes can ensure delivery of high concentrations at lesional sites and for sustained periods. Courses can be repeated. In addition to melanoma, cutaneous and bronchial squamous cell carcinoma, bladder and breast cancers, and leukaemia would seem to be ideal candidates for further clinical investigation and trial of the anti-cancer potential of azelaic acid, as prime, adjuvant, and palliative therapy, and for disseminated disease. PMID- 10362282 TI - Further appreciation of a control system for chemical reactions residing in virtual energy flows through the bio-system. AB - The argument has been advanced in previous papers that the radiative field of classical electromagnetics is a special case of envelopment of space itself (termed here 'non-reap' or 'imaginary energy') by waves of this field according to the formalism of Clark Maxwell (termed here 'real energy') and that, in its non-enveloped state, the virtual field is of special significance for the functioning bio-system. This paper summarizes evidence from a preceding paper from the areas of physical chemistry and theoretical physics to validate the presence of just such a radiationless, non-enveloped virtual field of space elements dissipating matter of inanimate and animate types. As discussed, the non enveloped field is subject to envelopment by magnetic fields and by the heat part of the electromagnetic spectrum. It is reasoned that the envelopment process itself is concerned with a thrust-delivering atomic and molecular movement so familiar in bio-system function. The virtual stream develops from the polyatom and polymer state so that wavelengths of its components are greater than those wavelengths associated with individual atoms or molecules. The division between the two wavelength types is set for purposes of this quasi-mechanical approach to space elements at the heat part of the spectrum. Properties of control and stability over atomic and molecular systems derive from this wavelength advantage. Irrespective of wavelength, the virtual system is operative free of heat and of the magnetic field. In the presence of appropriate threshold values, these enveloping agents then manifest an arrangement of space elements in real energy or radiative terms. Events during these transitions confer important physical properties on all matter but they are more highly developed and the control they exercise more subtle in animate matter. PMID- 10362283 TI - Does maximizing programmed cell death necessarily yield an optimum clinical advantage? AB - Generally it has been believed that a maximum therapeutic induction of programmed cell death in cancer cells is universally desirable. As a corollary, the presence of Bcl-2, a major anti-programmed cell death protein, is considered an unfavorable prognostic sign. The latter is not and the former may not be universally true. PMID- 10362284 TI - Three molecular mechanisms to explain some biological effects of electromagnetic fields and hypogravity. AB - There are many reports about the biological effects of electromagnetic fields and hypogravity and there have been many attempts to develop a theoretical explanation of this phenomenon. In this work, a mechanism is described based on the action of these physical environmental factors on single electrically charged groups from amino acids and considering the elongation stage of the protein synthesis as one of the main targets for both factors. For some rapid bioeffects after short exposures, a direct action on the conformation of the binding site of proteins is postulated. The other mechanism described here is based on the effect of these factors on the motion of the ionized calcium at the extracellular fluid. Many reports about the influences of electromagnetic and gravitational fields on gene expression, enzyme activity, bone mineralization, and oncogenesis are discussed, taking into account the new molecular mechanisms. PMID- 10362285 TI - Could bone marrow transplantation cure AIDS?: review. AB - Despite recent use of potent antiretroviral drugs, the goal of eradication of human immunodeficiency virus (HIV) and restitution of the immune system has not been achieved. The present work reviews the literature on syngeneic or allogeneic bone marrow transplantation (BMT) in patients infected with HIV and in patients with acquired immune deficiency syndrome (AIDS). Thirty-two such attempts have been reported between 1982 and 1996. In two cases, HIV was eradicated and previously positive polymerase chain reaction (PCR) for viral DNA and RNA became negative. One patient had transient disappearance of HIV by PCR and several more, including one who underwent xenotransplantation of baboon bone marrow, experienced clinical and laboratory improvement. These results correlate with use of intensive pre-transplant cytoablative conditioning with chemotherapy and radiotherapy. Curative mechanisms of conditioning include elimination of several cell populations: infected cells, cells susceptible to infection, uninfected chronically activated cells responsible for autoimmune phenomena, and exhausted haemopoietic and lymphopoietic progenitors. They are repopulated by donor-derived cells that could mount a successful antiviral response through cytotoxic T lymphocytes. Non-specific graft-versus-host effect of allogeneic bone marrow would also help eliminate residual reservoirs of virus, especially macrophages. Protection of repopulating cells from infection could be achieved by a combination of antiretroviral agents and gene therapy strategies may be of additional benefit. PMID- 10362286 TI - A role for oxygen-induced osmosis in hyperbaric oxygen therapy. AB - The principles of gas-induced osmosis, demonstrated in the 1970s, have been applied to the very large steady-state gradients of O2 arising between arterial blood and hypoxic tissue during hyperbaric oxygen (HBO) therapy to produce a fluid 'pump' in the desired direction for resolving accompanying oedema. Thus, in soft-tissue injuries, an oxygen-induced fluid pump would break the vicious cycle between ischaemia, hypoxia and oedema at the point of oedema rather than hypoxia, as hitherto assumed. This osmotic mechanism enables the successes of HBO therapy in hypoxic disorders to be reconciled with early failures in such areas as hyperbaric radiotherapy, where substitution of O2 for N2 in inspired air was clearly not reflected at the tissue level. This argument also applies to the success of HBO in treating air embolism and decompression sickness over simple compression. The oxygen pump would seem to offer a more plausible explanation for the success of HBO therapy than theories based upon O2 delivery by the circulation, especially when considering cardiovascular reflexes to elevated PaO2 and the marginal increase in blood O2 content upon switching to HBO from normobaric oxygen breathing. PMID- 10362287 TI - Mechanism of induction of follicular atresia after equine chorionic gonadotrophin (eCG) antisera treatment in hypophysectomized eCG-injected hamster model: possible involvement of c-myc and cdc25. AB - An intracellular mechanism of induction of apoptosis of granulosa cells implicated in the initiation of experimentally induced atresia of ovarian follicle in hypophysectomized eCG-injected hamster followed by eCG-antisera treatment is proposed. Induction of atresia after withdrawal of injected eCG by eCG-antisera treatment is possibly caused by inadequate level of the gonadotropin induced growth factor that results in apoptosis of granulosa cells associated with the activation of c-myc requiring cdc25A. PMID- 10362288 TI - Melatonin-dependent infertility. AB - Melatonin may be a key factor in the regulation of seasonal variation in gonadal activity. The circadian disturbances related to reproduction are probably subsequent to the seasonal change. Moreover, melatonin might also be considered essential for both spermatogenesis and folliculogenesis. Exposure to bright light, suppressing the concentration of melatonin in circulation, is hypothesized to be useful in treatment of both male and female infertility in couples with abnormal melatonin metabolism. PMID- 10362289 TI - Depletion in serotonin decreases neurogenesis in the dentate gyrus and the subventricular zone of adult rats. AB - During adulthood, neuronal precursor cells persist in two discrete regions, the subventricular zone and the hippocampal subgranular zone, as recently demonstrated in primates. To date, a few factors such as adrenal steroids and trophic factors are known to regulate adult neurogenesis. Since neuronal activity may also influence cellular development and plasticity in brain, we investigated the effects of serotonin depletion on cell proliferation occurring in these regions. Indeed, in addition to its role as a neurotransmitter, 5 hydroxytryptamine (serotonin) is considered as a developmental regulatory signal. Prenatal depletion in 5-hydroxytryptamine delays the onset of neurogenesis in 5 hydroxytryptamine target regions and 5-hydroxytryptamine promotes the differentiation of cortical and hippocampal neurons. Although in the adult brain, a few studies have suggested that 5-hydroxytryptamine may play a role in neuronal plasticity by maintaining the synaptic connections in the cortex and hippocampus, no information is actually available concerning the influence of 5 hydroxytryptamine on adult neurogenesis. If further work confirms that new neurons can be produced in the adult human brain as is the case for a variety of species, it is particularly relevant to determine the influence of 5 hydroxytryptamine on neurogenesis in the hippocampal formation, a part of the brain largely implicated in learning and memory processes. Indeed, lack of 5 hydroxytryptamine in the hippocampus has been associated with cognitive disorders, such as depression, schizophrenia and Alzheimer's disease. In the present study, we demonstrated that both inhibition of 5-hydroxytryptamine synthesis and selective lesions of 5-hydroxytryptamine neurons are associated with decreases in the number of newly generated cells in the dentate gyrus, as well as in the subventricular zone. PMID- 10362290 TI - Reduced chaos of interspike interval of midbrain dopaminergic neurons in aged rats. AB - In this study, the nonlinear prediction method combined with Gaussian-scaled surrogate data was used to quantify, as a first goal, the chaotic behavior of the interspike interval of ventral tegmental area dopaminergic neurons, extracellularly recorded in vivo, in anesthetized rats. The second goal was to determine the differences in chaotic content as a function of age. Comparisons were made among three different groups of rats: young (two to four weeks of age), adult (three to four months of age) and aged (16-19 months of age). It has been found that the degree of complexity of action potential trains is reduced with aging. The chaotic content of ventral tegmental area dopamine neurons within each group and the decrease of chaos with aging cannot be explained in terms of standard characteristics of neuronal activity (firing rate, bursting activity). These data can be rationalized in the light of recent findings on the role of deterministic chaos in the functional behavior of complex biological systems, and suggests that nonlinear analysis may provide an additional method in characterizing neuronal activity. PMID- 10362291 TI - The basal ganglia: a vertebrate solution to the selection problem? AB - A selection problem arises whenever two or more competing systems seek simultaneous access to a restricted resource. Consideration of several selection architectures suggests there are significant advantages for systems which incorporate a central switching mechanism. We propose that the vertebrate basal ganglia have evolved as a centralized selection device, specialized to resolve conflicts over access to limited motor and cognitive resources. Analysis of basal ganglia functional architecture and its position within a wider anatomical framework suggests it can satisfy many of the requirements expected of an efficient selection mechanism. PMID- 10362292 TI - Spontaneous activity of the perirhinal cortex in behaving cats. AB - The perirhinal cortex lies at the interface between the neocortex and allocortex. Whether the perirhinal cortex expresses spontaneous electroencephalographic rhythms that are characteristic of the allocortex and/or of the neocortex is unknown. Thus, the present investigation was undertaken to characterize the activity of the perirhinal cortex with respect to various electroencephalographic rhythms that are displayed by neocortical areas or the entorhino-hippocampal system during different behavioral states of vigilance in chronically-implanted cats. Although perirhinal and neocortical electroencephalograms underwent similar state-dependent changes in amplitude, the ubiquitous neocortical sleep spindles were absent from the perirhinal cortex. In addition, while the slow sleep oscillation (0.5-1 Hz), which is pervasive in the neocortex, was present in the perirhinal cortex, its temporal relation to the neocortical oscillation was highly variable. In contrast, a high degree of correlation was found between perirhinal and entorhinal electroencephalographic activities in all behavioral states. In particular, during waking and paradoxical sleep, multiple simultaneously recorded entorhinal and perirhinal sites displayed an oscillation in the theta range which was highly correlated. To rule out the possibility that the perirhinal theta oscillation reflected volume conduction from neighboring structures, single-unit recordings were performed. Spike-triggered averages and peri-event histograms revealed that perirhinal cells displayed a statistically significant theta-related modulation of their spontaneous activity, albeit weaker than that observed in the entorhinal cortex. Thus, from the standpoint of spontaneous electroencephalographic rhythms, the perirhinal cortex is more closely related to the entorhino-hippocampal system than to the neocortex. PMID- 10362293 TI - Alpha2-adrenoceptor modulation of cortical acetylcholine release in vivo. AB - Acetylcholine release in the rat cortex in vivo has been shown to be modulated by alpha2-adrenoceptor ligands. We have previously reported that the systemic administration of selective alpha2-antagonists including (+)-efaroxan increase, while alpha2-adrenoceptor agonists such as UK-14304 reduce the release of acetylcholine in the medial prefrontal cortex of conscious rats as measured by microdialysis. To evaluate the extent to which noradrenergic afferent inputs are required for the expression of these different effects, the present study examined the drug-induced changes in cortical acetylcholine release in rats which had undergone prior noradrenergic deafferentation. Rats were pretreated with the noradrenergic neurotoxin N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (40 mg/kg, i.p.), which after three days had reduced noradrenaline levels in the medial prefrontal cortex by 84%. At that time, slices of cortex were incubated with [3H]choline, superfused and stimulated by consecutive exposures to increasing concentrations of K+. In N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine pretreated tissue, the [3H] outflows evoked by 20, 35 and 45 mM K+ were lower by 12%, 22% and 43%, respectively, in comparison to slices prepared from vehicle-pretreated control animals. For in vivo microdialysis experiments, rats were pretreated as above with N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine, or prepared seven to eight days in advance with bilateral 6-hydroxydopamine lesions of the locus coeruleus. Neither of these lesioning procedures significantly affected the basal outflow of endogenous acetylcholine in the cortex. In control rats, cortical acetylcholine outflow was increased by up to 300% of baseline values by (+) efaroxan (0.63 mg/kg, i.p.), and was reduced to 21% of baseline by UK-14304 (2.5 mg/kg, i.p.), confirming our previous findings. In N-(2-chloroethyl)-N-ethyl-2 bromobenzylamine pretreated rats, the inhibitory effect of UK-14304 on acetylcholine outflow persisted, while the ability of (+)-efaroxan to increase outflow was essentially eliminated. In locus coeruleus-lesioned rats, where cortical noradrenaline levels were reduced by 64%, (+)-efaroxan still increased acetylcholine outflow, but this effect was significantly attenuated and less sustained in comparison to sham-operated control rats. Viewed together with complimentary biochemical, electrophysiological and neuroanatomical evidence in the literature, a model is presented to account for these findings, and indicates that alpha2-adrenoceptors both on noradrenergic neurons (autoreceptors) and on non-noradrenergic cells (heteroreceptors) can participate in mediating drug induced changes in medial prefrontal cortical acetylcholine release in vivo. The acetylcholine release-enhancing effect of (+)-efaroxan appears to be dependent on at least a partially intact cortical noradrenergic innervation. PMID- 10362294 TI - Involvement of interleukin-1beta in the mechanism of human immunodeficiency virus type 1 (HIV-1) recombinant protein gp120-induced apoptosis in the neocortex of rat. AB - The effect of subchronic intracerebroventricular injection of the human immunodeficiency virus type 1 (HIV-1) recombinant protein gp120 (100 ng, given daily for up to seven consecutive days) on interleukin-1beta expression was studied by immunohistochemistry in the brain of adult rats. In comparison to control, bovine serum albumin (300 ng, given intracerebroventricularly for up to seven days) -treated animals (n=6), interleukin-1beta immunoreactivity increased in the brain cortex and hippocampus of rats (n=6) receiving a single injection of the viral protein 24 h before analysis with more substantial increases being observed in these regions of the brain (n=6) after seven days treatment. Double labelling immunofluorescence experiments support a neuronal and, possibly, a microglial cell origin for gp120-enhanced interleukin-1beta expression. Transmission electron microscopy analysis of brain tissue sections revealed that combination treatments (given intracerebroventricularly daily for seven days) with gp120 (100 ng) and interleukin-1 receptor antagonist (80 ng) or with the interleukin converting enzyme inhibitor II (100 pmol), but not with leupeptin (100 pmol), prevented apoptotic death of rat (n=6/group) brain cortical cells typically elicited by the viral protein. These data demonstrate that gp120 enhances interleukin-1beta expression in the brain and this may be involved in the mechanism underlying apoptosis induced by gp120 in the brain cortex of rat. Further support to this hypothesis comes from the evidence that intracerebroventricular injection of murine recombinant interleukin-1beta (200 U, given daily for seven consecutive days) produces DNA fragmentation in the brain cortex of rat (n=6). Interestingly, the latter treatment enhanced nerve growth factor level in the hippocampus but not in the cerebral cortex and this coincides with a similar effect recently reported in identical brain areas of rats treated likewise with gp120. In conclusion, the present data demonstrate that treatment with gp120 enhances interleukin-1beta expression and this participates in the mechanism of apoptotic cell death in the brain cortex of rat. By contrast, in the hippocampus, gp120-enhanced interleukin-1beta expression elevates nerve growth factor that may prevent or delay apoptosis in this plastic region of the rat brain. PMID- 10362295 TI - Self-stimulation rewarding experience induced alterations in dendritic spine density in CA3 hippocampal and layer V motor cortical pyramidal neurons. AB - Self-stimulation rewarding experience induced alterations in the numerical density of spines in CA3 hippocampal and layer V motor cortical pyramidal neurons in adult male Wistar rats was evaluated. Self-stimulation experience was provided 1 h daily over a period of 10 days through stereotaxically implanted bipolar stainless steel electrodes bilaterally in lateral hypothalamus and substantia nigra-ventral tegmental area. After 10 days, rats were killed and the hippocampus and motor cortex were processed for rapid Golgi staining procedure. The dendritic spine densities were studied in CA3 hippocampal and layer V motor cortical pyramidal neurons. The spine densities were quantified in five successive segments of 15.2 microm up to a distance of 76 microm. Apical dendrites were classified as mainshaft, sub branch, oblique shaft-I, oblique shaft-II, primary branch; and basal dendrites as main shaft, primary branch and secondary branch. A grand total of 864 CA3 hippocampal and 1008 layer V motor cortical dendrites were analysed for spine counting in different groups of rats. The results revealed a significant (P<0.001; ANOVA, F-test) increase in the number of spines in all the categories of dendrites in apical and basal regions in both hippocampal and motor cortical neurons in self-stimulation group of rats. Such changes were not observed either in sham control, experimenter-administered or normal control groups of rats. The self-stimulation induced increase in the spine density suggests an increase in the postsynaptic receptive field in CA3 hippocampal and layer V motor cortical neurons. This might enhance the efficacy of synaptic transmission in these neurons. Our study clearly demonstrated the self stimulation rewarding experience induced postsynaptic plasticity in hippocampal and motor cortical pyramidal neurons. PMID- 10362296 TI - Ligand and subfield specificity of corticoid-induced neuronal loss in the rat hippocampal formation. AB - Adult male rats were treated chronically with the selective type II corticosteroid receptor agonist dexamethasone, with dexamethasone plus aldosterone, a selective type I receptor agonist, and with a supraphysiological dose of corticosterone sufficient to occupy both type I and type II receptors; injection-free and oil (vehicle)-treated rats served as controls. Following one month of treatment, the animals were killed and their brains were processed for stereological assessment of volumes and total numbers of neurons in the hippocampal formation. Dexamethasone treatment resulted in significant reductions in the total number of dentate granule and the CA3 pyramidal cells and in the volumes of some layers of these subfields; however, this steroid did not influence any morphometric parameter in the CA1 subfield, and the number of hilar cells was also unaltered. In contrast to the results obtained with dexamethasone, the other two groups of corticoid injected animals did not reveal changes in total cell numbers in any of the subfields of the hippocampal formation, although in the corticosterone-treated group a reduction in the volumes of the hilus and of the stratum radiatum of the CA3 subfield was observed. The present data show that the exclusive activation of type II corticosteroid receptors results in subfield-specific neuronal loss in the hippocampal formation of rats. This type II receptor-mediated neuronal loss can, however, be abrogated by the simultaneous stimulation of type I corticosteroid receptors. Together, these findings extend and support previous studies which suggested that activation of type I corticosteroid receptors may promote neuronal survival and that neurodegeneration may be triggered by type II corticosteroid receptor stimulation. An important implication of this result is that elevated levels of the endogenous corticosteroid receptor ligands (e.g., during stress) is unlikely to cause severe structural damage to the hippocampal formation due to the contemporaneous occupation of type I receptors. PMID- 10362297 TI - Expression of neurotrophins in hippocampal interneurons immunoreactive for the neuropeptides somatostatin, neuropeptide-Y, vasoactive intestinal polypeptide and cholecystokinin. AB - Using a double detection method, which combines in situ hybridization for the detection of neurotrophin messenger RNA with immunocytochemistry against the neuropeptides somatostatin, neuropeptide Y, vasoactive intestinal polypeptide and cholecystokinin, we have analysed the expression of the neurotrophins, nerve growth factor, brain-derived neurotrophic factor and neurotrophin-3, in distinct populations of neuropeptide-immunoreactive hippocampal interneurons. Nerve growth factor messenger RNA expression was found in subsets of the four subpopulations of neuropeptide-immunoreactive interneurons. The highest degree of co localization was observed in the neuropeptide-Y-positive cells (up to 70%) and in somatostatin-immunoreactive cells (48%). Only small subsets of cholecystokinin- and vasoactive intestinal polypeptide-positive neurons (21% and 10%, respectively) displayed nerve growth factor hybridization signals. In contrast, expression of neurotrophin-3 messenger RNA was exclusively observed in 26% of neuropeptide-Y-immunoreactive cells. Brain-derived neurotrophic factor hybridization signals were never detected in the neuropeptide-positive hippocampal interneurons. Morphological analysis of neuropeptide-immunoreactive interneurons that express or lack nerve growth factor messenger RNA revealed that most perisomatic inhibitory neurons, such as large vasoactive intestinal polypeptide/ cholecystokinin-immunoreactive cells, showed positive nerve growth factor hybridization signals. In addition, some somatostatin/neuropeptide-Y immunoreactive interneurons, which are responsible for dendritic inhibition of principal hippocampal neurons, expressed nerve growth factor messenger RNA. In contrast, interneurons specialized to innervate other GABAergic cells, such as small vasoactive intestinal polypeptide-positive cells, lacked nerve growth factor expression. All these data indicate that expression of neurotrophins is differentially regulated in functionally distinct classes of hippocampal interneurons immunoreactive for neuropeptides. We also analysed whether neuropeptide-immunoreactive interneurons expressing neurotrophins were targets of the GABAergic septohippocampal pathway. We used a triple detection method, combining anterograde tracing of this connection, with in situ hybridization for the detection of neurotrophin mRNA, and immunocytochemistry against neuropeptides. Our data showed that the four populations of hippocampal interneurons studied (somatostatin, neuropeptide-Y, vasoactive intestinal polypeptide and cholescystokinin) received GABAergic afferents from the septum. However, no preference for neuropeptide-immunoreactive cells expressing neurotrophins was observed, compared to neuropeptide-positive neurons lacking neurotrophin expression. PMID- 10362298 TI - Taurine release modified by nitric oxide-generating compounds in the developing and adult mouse hippocampus. AB - The effects of the nitric oxide-generating compounds hydroxylamine, sodium nitroprusside and S-nitroso-N-acetylpenicillamine, and the nitric oxide synthase inhibitors nitroarginine and 7-nitroindazole on taurine release from hippocampal slices from adult (three-month-old) and developing (seven-day-old) mice were characterized using a superfusion system. The basal release of [3H]taurine was enhanced when the nitric oxide donors were added at the beginning of superfusion, more markedly in the adult than in the immature hippocampus. The effect of hydroxylamine was clearly concentration-dependent. Hydroxylamine also markedly enhanced the release of endogenous taurine. The K+-stimulated (50 mM) release of taurine was generally inhibited by the nitric oxide-generating compounds in both age groups. Nitric oxide is thus able to act directly at presynaptic terminals, modulating taurine release as a retrograde messenger. The N-methyl-D-aspartate evoked taurine release was reduced by the nitric oxide donors, particularly by sodium nitroprusside, in the adult hippocampus, while hydroxylamine and S-nitroso N-acetylpenicillamine markedly potentiated the release in developing mice. In the immature hippocampus the hydroxylamine-enhanced taurine release seems to involve a Ca2+-independent, Na+-dependent and carrier-mediated process while in adult mice only a part of the hydroxylamine-enhanced release is mediated by the same mechanism. The results show that nitric oxide-generating compounds modify the basal, K+- and N-methyl-D-aspartate-evoked releases of taurine in both adult and immature hippocampus. The enhanced N-methyl-D-aspartate receptor-evoked release may be an important mechanism protecting the immature brain against excitotoxicity. PMID- 10362300 TI - Leukemia inhibitory factor regulates galanin/galanin message-associated peptide expression in cultured mouse dorsal root ganglia; with a note on in situ hybridization methodology. AB - After transection of the sciatic nerve there is a dramatic increase in both galanin/galanin message-associated peptide-like immunoreactivities and preprogalanin messenger RNA levels in rat and mouse lumbar 4 and 5 dorsal root ganglion neurons. There is strong evidence that after nerve injury leukemia inhibitor factor is a key molecule in the control of peptide expression both in sympathetic neurons and in dorsal root ganglion neurons, although the cells of origin of endogenous leukemia inhibitory factor remain to be established. We have therefore studied the effect of leukemia inhibitory factor on galanin expression in 72 h cultured dorsal root ganglion neurons from normal mice, leukemia inhibitory factor-deficient and heterozygous mice with immunohistochemistry and in situ hybridization. In cultures of leukemia inhibitory factor-deficient (-/-) mice only 13% of the dorsal root ganglion neurons expressed galanin message associated peptide and in cultures from heterozygous (+/-) and wild-type (+/+) mice the corresponding figures were, respectively, 24 and 40%. After addition of leukemia inhibitory factor (10 or 50 ng/ml) to the culture medium, the number of neurons expressing galanin message-associated peptide was increased (up to 41%) in cultures from (-/-) animals after the high concentration and reached similar values in cultures from heterozygous animals incubated with the low concentration. These findings were supported by parallel analysis of prepro galanin messenger RNA levels, where similar transcript levels and effects in the various cultures were observed in the non-radioactive in situ hybridization experiments. These results support the hypothesis that leukemia inhibitory factor is an important regulator of galanin/galanin message-associated peptide expression following axotomy, and may therefore be involved in the defence mechanisms against neuropathic pain at the level of dorsal root ganglion neurons. PMID- 10362299 TI - Leukaemia inhibitory factor prevents loss of p75-nerve growth factor receptor immunoreactivity in medial septal neurons following fimbria-fornix lesions. AB - Transection of the fimbria-fornix leads to retrograde degeneration of axotomized septal cholinergic neurons as manifested by loss of choline acetyltransferase and low-affinity nerve growth factor receptor (p75NGFR) immunoreactivity. Nerve growth factor administered into cerebral ventricles at the time of axotomy can prevent these changes, while ciliary neurotrophic factor can prevent the loss of p75NGFR immunostaining. Leukaemia inhibitory factor shares structural homologies with ciliary neurotrophic factor and has similar actions in the nervous system. Both proteins share the same signalling pathways, which involve the interleukin-6 transducing receptor components leukaemia inhibitory factor receptor beta and gp130. In this study, we compared the effects of leukaemia inhibitory factor, ciliary neurotrophic factor and nerve growth factor, administered into cerebral ventricles, on p75NGFR and choline acetyltransferase immunoreactivity in septal neurons after fimbria-fornix transection. We found that leukaemia inhibitory factor, like ciliary neurotrophic factor, prevents the loss of p75NGFR-stained medial septal neurons after fimbria-fornix axotomy, without maintaining choline acetyltransferase expression in these neurons. In addition, p75NGFR-immunostained neurons had significantly smaller mean diameter after axotomy in leukaemia inhibitory factor- and ciliary neurotrophic factor-treated animals as compared with either nerve growth factor-treated or unlesioned animals. These findings suggest that both leukaemia inhibitory factor and ciliary neurotrophic factor can prevent the axotomy-induced cell death of septal cholinergic neurons, but that, in contrast to nerve growth factor, these growth factors do not maintain the expression of choline acetyltransferase or the normal neuronal size of these injured neurons. PMID- 10362301 TI - Serotonin and acetylcholine release response in the rat hippocampus during a spatial memory task. AB - By using in vivo microdialysis we monitored the extracellular levels of acetylcholine and serotonin in the hippocampus of rats performing a spatial memory task. After rats were trained for 10 consecutive days to master a food reinforced radial-arm maze task, they were implanted with a microdialysis probe in the dorsal hippocampus. On day 12, rats were tested in the maze and acetylcholine and serotonin outputs were monitored before the test, during the waiting phase and while performing the trials. In trained, food-rewarded rats, hippocampal acetylcholine levels increased during the waiting period (181 +/- 90 of baseline) and further increased during the radial-maze performance to 236 +/- 13% of baseline values, while serotonin levels did not change during the waiting period but increased to 142 +/- 3% during the maze performance. To discriminate whether the increase of acetylcholine and serotonin levels during the testing was associated with memory performance or with food consumption, we monitored hippocampal acetylcholine and serotonin release in rats that were trained, but not food rewarded, or in rats that were not trained, but rewarded only on the test day. In the trained, non-rewarded group, acetylcholine release increased during the waiting phase to 168 +/- 6%, but did not increase further during the task performance. In contrast, no change in serotonin release was observed in this group in any phase of the test. In rats which were not trained, but food rewarded, acetylcholine increased only during the maze period (150 +/- 5%). Serotonin increased gradually and become significant at the end of the trials. (130 +/- 3%). While both neurotransmitters could be implicated in feeding behaviour, only activation of cholinergic neurotransmission appears to be associated with memory function. Our results support the following hypotheses: (i) hippocampal acetylcholine could be involved in attentional and cognitive functions underlying motivational processes; (ii) serotonin could be implicated in non-cognitive processes (i.e. in the control of motor and feeding behaviour). Since serotonin and acetylcholine neurotransmission is simultaneously activated during the spatial memory task, this suggests that these neurotransmitter systems regulate behavioural and cognitive functions. PMID- 10362302 TI - Learning-induced plasticity of N-methyl-D-aspartate receptors is task and region specific. AB - Changes in binding of [3H]dizocilpine maleate to N-methyl-D-aspartate-sensitive ion channel receptors were evaluated after learning in order to specify brain regions which might be involved in memory formation. Rats were trained in a five trial session of 40 min, to discriminate among three odours to obtain food reinforcement. Another group was trained in an eight-arm maze to choose always the same three arms to obtain food reinforcement (nine trials over 150 min). In rats killed 30 min after odour discrimination learning, dizocilpine maleate binding was significantly reduced in hippocampal sub-regions CA3, CA1 and fascia dentata and in frontal cortex. After spatial learning, changes in binding were limited to the amygdala, where a decrease was also observed. These results indicate that functional changes occur in specific brain regions after learning and suggest anatomical loci for further study of synaptic changes at a morphological level, after spatial learning or odour discrimination. PMID- 10362303 TI - Nitric oxide synthase (NADPH-diaphorase) content in brain neurons of neonatal rats after inhibitory learning and intervention into nitric oxide metabolism. AB - Four-day-old rat pups were taught to avoid an electrified grid under the influence of increased nitric oxide availability in brain (by a nitric oxide substrate L-arginine) that alleviated learning or decreased nitric oxide (due to the action of a blocker of nitric oxide synthase nitro-L-arginine) that impaired learning. Three hours after criteria meeting, the pups were killed for analysis of nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase content in brain cells and neuropil. In the cingulate gyrus, NADPH-diaphorase-positive staining was increased after L-arginine, but an opposite picture was observed in hippocampus and basal ganglia, i.e. an increase after the blocker nitro-L arginine. A noteworthy accumulation of NADPH-diaphorase in hippocampal cells might be tentatively explained by the blocking effect of nitro-L-arginine not allowing NADPH-diaphorase to leave the cells. Application of L-arginine or nitro L-arginine provoked only minor changes in the studied structures of non-learned pups with the exception of hippocampus where nitro-L-arginine increased the width of neuropil, but to a lesser degree than in learned animals. These results clearly show that both manipulations, i.e. drug application and learning, only have a significant effect on the changes in NADPH-diaphorase positivity in brain neurons. PMID- 10362304 TI - Forskolin and dopamine D1 receptor activation increase huntingtin's association with endosomes in immortalized neuronal cells of striatal origin. AB - Huntingtin is a cytoplasmic protein of unknown function that associates with vesicle membranes and microtubules. Its protein interactions suggest that huntingtin has a role in endocytosis and organelle transport. In this study we sought to identify factors that regulate the transport of huntingtin in striatal neurons, which are the cells most affected in Huntington's disease. In clonal striatal cells derived from fusions of neuroblastoma and embryonic striatal neurons, huntingtin localization is diffuse and slightly punctate in the cytoplasm. When these neurons were differentiated by treatment with forskolin, huntingtin redistributed to perinuclear regions, discrete puncta along plasma membranes, and branch points and terminal growth cones in neurites. Huntingtin staining overlapped with clathrin, a coat protein involved in endocytosis. Immunoblot analysis of subcellular membrane fractions separated by differential centrifugation confirmed that huntingtin immunoreactivity in differentiated neurons markedly increased in membrane fractions enriched with clathrin and with huntingtin-interacting protein 1. Dopamine treatment altered the subcellular localization of huntingtin and increased its expression in clathrin-enriched membrane fractions. The dopamine-induced changes were blocked by the D1 antagonist SCH 23390 and were absent in a clonal cell line lacking D1 receptors. Results suggest that the transport of huntingtin and its co-expression in clathrin and huntingtin-interacting protein 1-enriched membranes is influenced by activation of adenylyl cyclase and stimulation of dopamine D1 receptors. PMID- 10362305 TI - N-methyl-D-aspartate receptor blockade affects polysialylated neural cell adhesion molecule expression and synaptic density during striatal development. AB - Glutamatergic neurons innervate the striatum and form asymmetric synapses with the dendritic spines of striatal efferent neurons. The role of glutamate in striatal development, however, remains largely unknown. Previous studies have shown a dramatic increase in the density of asymmetric synapses in the rat striatum during the third postnatal week, followed by a decrease to adult levels by postnatal day 25. At the same time, the highly polysialylated form of the neural cell adhesion molecule becomes progressively restricted to synaptic regions and then disappears. We have now examined the effects of antagonists of the N-methyl-D-aspartate subtype of glutamatergic receptors on the expression of the polysialylated form of the neural cell adhesion molecule and on synaptic density during this late period of striatal development. Peripheral administration of the N-methyl-D-aspartate receptor antagonist dizocilpine maleate markedly decreased immunoreactivity for the highly polysialylated form of the neural cell adhesion molecule in the dorsolateral striatum and cerebral cortex when drug treatment included postnatal day 20, but not earlier in development. This effect was regionally specific and loss of the polysialylated neural cell adhesion molecule in the striatum was reproduced by the local administration of dizocilpine maleate, DL-2-amino-5-phosphonovalerate or ketamine on postnatal day 20. Quantitative ultrastructural studies of synaptic density with the physical disector method performed after one of the regimens inducing loss of the polysialylated neural cell adhesion molecule (postnatal days 18-20) revealed a 30% decrease in asymmetric synapses in the dorsolateral striatum of treated rats. Symmetric synapses, which presumably do not use glutamate, were not affected. The data indicate that N-methyl-D-aspartate receptors play a role in the late stages of synaptogenesis in the striatum and suggest that a subset of synapses expressing immunoreactivity for the highly polysialylated form of the neural cell adhesion molecule may be dependent on N-methyl-D-aspartate receptor stimulation during a critical period of striatal development. PMID- 10362306 TI - Increased striatal expression of glutamate decarboxylase 67 after priming of 6 hydroxydopamine-lesioned rats. AB - Previous single exposure (priming) to a dopamine receptor agonist greatly enhances the contralateral turning behaviour elicited by dopamine D1 receptor agonists in unilaterally 6-hydroxydopamine lesioned rats. In the present study we have investigated the levels of glutamate decarboxylase 67 and glutamate decarboxylase 65 messenger RNA in the striatum of 6-hydroxydopamine-lesioned rats primed with L-3,4-dihydroxyphenylalanine (L-DOPA) and challenged with the D1 receptor agonist SKF 38393, three days thereafter. As previously reported, levels of glutamate decarboxylase 67 messenger RNA increased in the striatum denervated by the 6-hydroxydopamine lesion as compared with the intact one. Striatal glutamate decarboxylase 67 messenger RNA levels, measured three days after priming with L-DOPA (50 mg/kg), further increased in the lesioned striatum while were not modified in the intact one. Administration of SKF 38393 (3 mg/kg) elicited a more intense contralateral turning behaviour in primed than in drug naive 6-hydroxydopamine-lesioned rats but did not induce any change in striatal glutamate decarboxylase 67 messenger RNA. In contrast, striatal levels of glutamate decarboxylase 65 messenger RNA were not modified by either 6 hydroxydopamine lesions or priming with L-DOPA. The results show that priming with L-DOPA induces long-lasting changes in GABAergic neurons of the 6 hydroxydopamine-lesioned striatum. These changes might play a role in the increased behavioural response of striatal D1 receptors induced by priming. PMID- 10362307 TI - Thalamic inputs to striatal interneurons in monkeys: synaptic organization and co localization of calcium binding proteins. AB - Recent studies indicate that extrinsic inputs from sensorimotor regions of the cerebral cortex and the centromedian intralaminar thalamic nucleus terminate preferentially upon specific subpopulations of striatal output neurons in monkeys. The objective of the present study was to verify whether this specificity of innervation also characterizes the synaptic interactions between thalamic inputs from the centromedian nucleus and the four major populations of striatal interneurons. This was achieved by double labelling techniques at the electron microscope level, combining the anterograde transport of biotinylated dextran amine with the immunostaining for specific markers of striatal interneurons (somatostatin, parvalbumin, choline acetyltransferase and calretinin). Injections of biotinylated-dextran amine in the centromedian nucleus led to dense bands of anterograde labelling which, in double immunostained sections, largely overlapped with the four populations of interneurons in the post-commissural region of the putamen. In the electron microscope, biotinylated dextran amine-containing terminals formed asymmetric axo-dendritic synapses with somatostatin-, parvalbumin-, and choline acetyltransferase-containing elements. However, synapses between anterogradely labelled terminals and calretinin positive neurons were not found. In sections processed to localize biotinylated dextran amine and parvalbumin or calretinin, double-labelled terminals (biotinylated-dextran amine/parvalbumin and biotinylated-dextran amine/calretinin), morphologically similar to thalamostriatal boutons, were found in the striatum indicating that calcium binding proteins may be expressed by thalamostriatal neurons. To test this possibility, we combined the retrograde transport of lectin-conjugated horseradish peroxidase from the putamen with parvalbumin and calretinin immunostaining and found that, indeed, most of the retrogradely labelled cells in the centromedian nucleus displayed parvalbumin and calretinin immunoreactivity. Moreover, co-localization studies revealed that calretinin and parvalbumin co-exist in single neurons of the centromedian nucleus. In conclusion, striatal interneurons immunoreactive for somatostatin, parvalbumin and choline acetyltransferase, but not those containing calretinin, receive strong inputs from the centromedian nucleus in monkeys. Moreover, our findings indicate that parvalbumin and calretinin co-exist in individual thalamostriatal neurons. In combination with our previous data, these results suggest that thalamic information may be conveyed to striatal projection neurons both, directly via excitatory synaptic inputs, or indirectly via striatal interneurons. The relative importance of those direct and indirect thalamic influences upon the activity of striatal output neurons remains to be established. PMID- 10362308 TI - Different sensitivity of in vivo acetylcholine transmission to D1 receptor stimulation in shell and core of nucleus accumbens. AB - We investigated whether D1 dopaminergic receptors modulate in vivo acetylcholine output in the shell and core areas of rat nucleus accumbens using the microdialysis technique. Subcutaneous injection (1, 2 and 3 mg/kg) of the D1 agonist SKF 82958 enhanced acetylcholine output in both areas of the nucleus accumbens while the selective D1 antagonist SCH 39166 (0.15 and 0.30 mg/kg, s.c.) lowered it. Both SKF 82958 and SCH 39166 were more effective in the shell than in the core region. The increase in acetylcholine release induced by SKF 82958 in the shell was tetrodotoxin-sensitive. The dopamine release inducer d-amphetamine (1 and 2mg/kg, s.c.) and the dopamine uptake inhibitor cocaine (10 and 20 mg/kg, i.p.) dose-dependently raised acetylcholine release in the shell and core areas. The dopaminergic stimulants, like the direct-acting D1 compounds, were more effective in the shell than in the core compartment of the nucleus accumbens. The acetylcholine increases in the shell induced by d-amphetamine (2 mg/kg), cocaine (20 mg/kg) and SKF 82958 (3 mg/kg) were antagonized by the D1 antagonists SCH 39166 (5 microM) and SCH 23390 (10 microM), applied locally by reverse dialysis. The results suggest that dopamine acting at the D1 receptors exerts a tonic stimulatory control over the cholinergic function of the shell and core compartments of the nucleus accumbens with the shell being more strongly influenced. PMID- 10362309 TI - Endogenous opioids implicated in the dynamics of experimental drug addiction: an in vivo autoradiographic analysis. AB - Endogenous opioids have been implicated in the neurobiological mechanisms underlying drug addiction. Although some information is available concerning effects of abused drugs on the endogenous opioid systems, the interpretation of these effects is hampered because data on the actual changes in the endogenous opioids during the dynamics of the drug addiction are lacking. The present report deals with changes in endogenous opioid activity before and after the daily self administration session in rats offered cocaine or ethanol, using an in vivo autoradiographic receptor occupancy procedure. In separate saline-controlled experiments drug-naive rats were allowed to intravenously self-administer cocaine (30 microg/infusion) and ethanol (0.05%) for five consecutive daily sessions of 6 h. Immediately following the last session on day 5 or just before a scheduled next daily session on day 6, the rats were injected with [3H]diprenorphine and subsequently prepared for autoradiography. Decreased [3H]diprenorphine binding was observed throughout the subcortical brain after the daily session in cocaine, but hardly in animals self-administering ethanol. These changes are thought to reflect a direct or an indirect effect of the drug on endogenous opioid systems. Before the daily session, the [3H]diprenorphine binding was decreased in restricted areas of the mesocorticolimbic system and of the thalamus in both cocaine and ethanol self-administering animals. These data suggest that release of endogenous opioids at the time the desire for cocaine or ethanol is high, which may be pertinent for drug-induced craving and relapse of drug addicts. PMID- 10362310 TI - Increased activity in the form of endurance training increases calcitonin gene related peptide content in lumbar motoneuron cell bodies and in sciatic nerve in the rat. AB - The relative content of calcitonin gene-related peptide in lumbar motoneuron cell bodies (semiquantitative immunohistochemistry) and sciatic nerve was examined in rats who had previously undergone a 16-week period of endurance training on a motor-driven treadmill. Soleus motoneurons were identified in the spinal cord by their fluorescence following injection of FluoroGold into the muscle one week before killing. In sedentary rats, calcitonin gene-related peptide was detectable in 76-90% of motoneurons, with no difference in the proportions of negative cells, or in the mean staining intensity of positive cells, between soleus and neighbouring (presumptive fast hindlimb muscle) unlabelled moto-neurons. In endurance-trained rats, the estimated content of calcitonin gene-related peptide was significantly increased (90%) in cell bodies of soleus and neighbouring motoneurons, with no training-induced alterations in the proportions of calcitonin gene-related peptide-positive cells in either sample. The content of the neuropeptide was also significantly higher (37%) in sciatic nerve of endurance-trained rats. Relative accumulation of calcitonin gene-related peptide proximal to a sciatic nerve ligature applied 4 h before, however, was unchanged. The increases in calcitonin gene-related peptide in motoneuron cell bodies and sciatic nerve axons following endurance training may indicate an up-regulation of the synthesis, transport and terminal release of this neuropeptide, which could play a significant role in other morphological and functional adaptations which are known to occur at the neuromuscular junction following this chronic change in activity level. PMID- 10362311 TI - Hypothalamic serotonergic activity correlates better with brain temperature than with sleep-wake cycle and muscle tone in rats. AB - The activity of the serotonergic system varies in phase with the sleep-wake cycle, which is associated with changes in several physiological functions, including electroencephalographic activity, brain temperature, and locomotion. The aim of the present study was to clarify which of these parameters correlates better with serotonergic activity in spontaneous conditions. Voltammetric recordings by telemetry of serotonergic metabolism in the medial preoptic area and polygraphic recordings of sleep-wake activity (by means of electroencephalographic delta band, brain cortical temperature and neck electromyographic activity recordings) were simultaneously performed in freely moving rats. Univariate analyses of variance revealed that each variable under investigation was statistically correlated with serotonergic metabolism. When the variables were entered into the model simultaneously, both partial correlation and step-wise multiple regression analyses indicated that the highest correlation exists between serotonergic metabolism and brain cortical temperature. The present data show that serotonergic activity in the medial preoptic area is closely linked to physiological changes in brain temperature. PMID- 10362312 TI - Tonic adrenergic and serotonergic inhibition of a withdrawal reflex in rabbits subjected to different levels of surgical preparation. AB - The excitability of the heel-gastrocnemius withdrawal reflex pathway has been monitored in rabbits undergoing surgical preparation for electrophysiological experimentation under Saffan anaesthesia. Reflexes were evoked by percutaneous electrodes inserted at the heel and recorded as electromyograph signals from the ipsilateral medial gastrocnemius muscle. Two levels of surgery were carried out. The "full surgical" preparation was performed under deep Saffan anaesthesia. The trachea, carotid artery, jugular vein and intrathecal space (via a small laminectomy at L1) were cannulated, the animals were decerebrated by suction, and the left hindlimb was immobilized by screw clamps applied to the tibia and the femur. The sciatic nerve and its branches were exposed by bisection of the posterior biceps muscle and the anaesthetic was withdrawn. In the "reduced surgery" preparation, procedures were carried out with a lighter level of Saffan anaesthesia and operated tissues were infiltrated with local anaesthetic. Only the cannulations were performed in these animals. The excitability of the heel gastrocnemius reflex declined throughout the full surgical preparation, with the median threshold increasing from 0.8 to 4.2 mA (n=19) and responses to suprathreshold stimuli reducing in size. Most of this effect was reversed after surgery was complete and anaesthesia withdrawn subsequent to decerebration. There were no significant changes in reflex excitability during the reduced surgery preparation (n = 15). Animals prepared by each of these protocols were given increasing intrathecal doses of either the selective alpha2-adrenoceptor antagonist RX 821002 (0.3 to 300 microg) or the serotonin/5-hydroxytryptamine (5 HT)1A-receptor antagonist WAY-100635 (0.01 to 30 microg). Both drugs caused significant, dose-dependent increases in reflex responses, to four to six times pre-drug control in both groups of animals. There were no differences in the effects on reflexes of either drug between the preparations. Thus, surgical preparation of decerebrated rabbits for electrophysiological recording results in depression of hindlimb withdrawal reflexes, although much of this effect did not persist beyond the completion of surgery. Tonic monoaminergic inhibition of reflexes was present to the same extent in both preparations investigated and is not therefore an epiphenomenon of the way in which the animals were prepared. PMID- 10362313 TI - Involvement of neurokinins in antidromic vasodilatation in hairy and hairless skin of the rat hindlimb. AB - By intravenous application of the specific neurokininl receptor antagonist SR 140333 and the specific calcitonin gene-related peptide receptor antagonist CGRP8 37 we tested to what extent neurokinins (substance P, neurokinin A) and calcitonin gene-related peptide are involved in mediating antidromic vasodilatation in skin of anaesthetized Wistar rats. The lumbar sympathetic chain was sectioned bilaterally between ganglia L2 and L3 to remove ongoing vasoconstrictor activity to the hindquarter. The left dorsal root L5 was stimulated electrically at 1 Hz with 20 pulses supramaximal for activating C fibres to evoke antidromic vasodilatation which was measured with laser Doppler flowmetry on the glabrous plantar skin and the hairy skin of the lower hindlimb within the left L5 territory. Stimulation-induced vasodilatation was tested after applying SR 140333 (0.1 mg/kg) and CGRP8-37 (0.3 mg/kg) alone or in combination. SR 140333 delayed the onset of the vasodilatation, but did not change its amplitude. CGRP8-37 reduced the amplitude and duration of the vasodilatation, but did not affect the latency of its onset. In combination, SR 140333 potentiated the effect of CGRP8-37 on the amplitude of the vasodilatation in glabrous but not in hairy skin and CGRP8-37 potentiated the delayed onset produced by SR 140333 in both cutaneous tissues. Antidromic vasodilatation in glabrous skin was almost totally blocked by SR 140333 (0.1 mg/kg) in combination with CGRP8-37 (0.45 mg/kg), but a substantial dilatation remained in hairy skin. It is concluded that in rat glabrous skin the vasodilatation evoked by a low level of activity in small-diameter primary afferents is likely to result from the release and synergistic action of neurokinins (substance P and/or neurokinin A) and calcitonin gene-related peptide, while in hairy skin neurokinins are involved to a minor extent only. PMID- 10362315 TI - Differential expression of retinoid receptors in the adult mouse central nervous system. AB - The immunocytochemical distribution of retinoid receptors has been analysed in the mouse adult central nervous system. All retinoic acid receptors (alpha, beta and gamma) and retinoid X receptors (alpha, beta and gamma) were detected and found to exhibit specific patterns of expression in various areas of the telencephalon, diencephalon and rhombencephalon. The protein localization of several retinoic acid receptors and retinoid X receptors did not correlate with the distribution of the corresponding RNA transcripts, as studied by in situ hybridization and RNase protection assays. This suggests that the expression of retinoid receptors could be post-transcriptionally regulated, which may contribute to their specific localization in the adult nervous system. PMID- 10362314 TI - Neurokinin-3 receptor distribution in rat and human brain: an immunohistochemical study. AB - Autoradiographic and immunohistochemical studies have shown that the neurokinin-3 receptor is widely distributed in the rodent CNS. Expression of the neurokinin-3 receptor in human brain, however, has been debated. These conflicting findings, as well as the poor resolution of autoradiographic images, prompted us to develop a polyclonal antibody against an oligopeptide derived from the carboxy-terminus consensus sequence of both the rat and human neurokinin-3 receptor ([C]ASTTSSFISSPYTSVDEYS, amino acids 434-452 of the rat neurokinin-3 receptor). Western blot analysis of both human and rat brain tissue revealed a major band in the molecular weight range 65,000-67,000, the proposed molecular weight of the neurokinin-3 receptor based on its amino acid sequence and presumed glycosylation state. The distribution of selective high affinity neurokinin-3 receptor agonist [3H]senktide binding and neurokinin-3 receptor immunoreactivity were virtually identical in the brains of male Fischer 344 rats. The highest concentrations of neurokinin-3 receptors were observed in cortical layers IV-V; the basolateral amygdaloid nucleus; the hypothalamic paraventricular, perifornical and supraoptic nuclei; the zona incerta; and the entopeduncular and interpeduncular nuclei. [3H]senktide binding and neurokinin-3 receptor immunoreactivity were compared in homologous cortical areas of the human and rat brain. In contrast to the rat, autoradiographic analysis of normal control human brains (35-75 years) revealed a distinct and predominant superficial cortical labeling in the glia limitans and the cortical layer I. However, neurokinin-3 receptor immunoreactivity could be found not only in the superficial cortical layers, but also on pyramidal neurons and astrocytes in the neuropil and white matter. These findings suggest species differences in both the cellular and anatomical distribution of the neurokinin-3 receptor. PMID- 10362316 TI - Carboxypeptidase D is a potential candidate to carry out redundant processing functions of carboxypeptidase E based on comparative distribution studies in the rat central nervous system. AB - Post-translational processing is essential for the biological activation of many proteins and peptides. After precursor cleavage at specific single residues or pairs of basic residues by the proprotein convertases, the C-terminal basic residues are removed. Carboxypeptidase E was thought to be the only enzyme responsible. Recent studies with carboxypeptidase E-deficient mice, Cpe(fat)/Cpe(fat), indicated the existence of carboxypeptidase E-like carboxypeptidases, such as carboxypeptidase D. In order to define potential redundant functions in vivo, we compared the distributions of both carboxypeptidases in the rat central nervous system and selected endocrine tissues. Carboxypeptidase D messenger RNA was abundantly expressed in glial cells in the gray and white matter, while neurons in several brain regions, such as the piriform cortex, basolateral amygdala and hippocampus, also expressed high levels of carboxypeptidase D messenger RNA. Co-localization of carboxypeptidases E and D messenger RNAs was observed in many brain regions, the spinal cord and endocrine tissues. Immunohistochemistry showed the intracellular distribution of carboxypeptidase D with a perinuclear pattern. The extensive distribution of carboxypeptidase D in both glial and neuronal cells indicates the important role of carboxypeptidase D in peptide processing, possibly working together with furin, a ubiquitously expressed proprotein convertase. The co-localization of carboxypeptidases D and E suggests that carboxypeptidase D may, at least partially, compensate for carboxypeptidase E processing functions in Cpe(fat)/Cpe(fat) mice. PMID- 10362317 TI - Distribution of neurons projecting to the rostral ventrolateral medullary pressor region that are activated by sustained hypotension. AB - Hypotension produces a reflex increase in the activity of sympathetic vasomotor and cardiac nerves. It is believed that the reflex sympathoexcitation is due largely to disinhibition of sympathoexcitatory neurons in the rostral ventrolateral medulla, but it is possible that it may also be mediated by excitatory inputs from interneurons that are activated by a fall in blood pressure. The aim of this study in conscious rabbits was to identify and map neurons with properties that are characteristic of interneurons conveying excitatory inputs to the rostral ventrolateral medullary pressor region in response to hypotension. In a preliminary operation, a retrogradely-transported tracer, fluorescent-labelled microspheres, was injected into the functionally identified pressor region in the rostral ventrolateral medulla. After a waiting period of at least one week, a moderate hypotension (decrease in arterial pressure of approximately 20 mmHg) was induced in conscious rabbits for 60 min by the continuous infusion of sodium nitroprusside. In confirmation of a previous study from our laboratory, [Li and Dampney (1994) Neuroscience 61, 613634] hypotension resulted in the expression of Fos (the protein product of c-fos, a marker of neuronal activation) in many neurons in several distinct regions in the brainstem and hypothalamus. Some of these regions (nucleus tractus solitarius, area postrema, caudal and intermediate ventrolateral medulla, parabrachial complex in the pons, and paraventricular nucleus in the hypothalamus) also contained large numbers of retrogradely-labelled cells. Approximately 10% of the Fos-positive neurons in the nucleus tractus solitarius, and 15-20% of Fos positive neurons in the caudal and intermediate ventrolateral medulla were also retrogradely-labelled from the rostral ventrolateral medullary pressor region. In other brain regions, very few double-labelled neurons were found. In previous studies from our laboratory, we have determined the distribution of neurons in the brainstem that project to the rostral ventrolateral medullary pressor region and that are also activated by hypertension [Polson et al. (1995) Neuroscience 67, 107-123] or by hypoxia. [Hirooka et al. (1997) Neuroscience 80, 1209-1224] Comparison of the present results with those from these previous studies indicate that although hypotension and hypoxia both elicit powerful reflex sympathoexcitatory responses, the central pathways subserving these effects in conscious animals are fundamentally different. Hypoxia activates rostral ventrolateral medullary sympathoexcitatory neurons mainly via a major direct excitatory projection from the nucleus tractus solitarius, as well as from the Kolliker-Fuse nucleus in the pons, while in contrast the activation of these neurons in response to hypotension appears to be due mainly to disinhibition, mediated via inhibitory interneurons. In addition, however, inputs originating from excitatory interneurons in the nucleus tractus solitarius and caudal and intermediate parts of the ventrolateral medulla appear to contribute to the hypotension-evoked activation of sympathoexcitatory neurons in the rostral ventrolateral medulla. PMID- 10362318 TI - Medial prefrontal cortex depressor response: role of the solitary tract nucleus in the rat. AB - The depressor response elicited by unilateral low intensity electrical stimulation of the rat ventral medial prefrontal cortex may be mediated by a connection with the solitary tract nucleus. We tested this hypothesis by (i) examining the influence of medial prefrontal cortex stimulation on the induction of Fos-like immunoreactivity in neurons in the medulla oblongata, and (ii) by testing the effect of inhibition of solitary tract nucleus neurons on the medial prefrontal cortex stimulation-evoked depressor response. Depressor responses (>10 mmHg) were elicited by electrical stimulation of the medial prefrontal cortex every minute for 1 h ('Stimulated' group). Control animals were treated identically but did not receive electrical stimulation ('Unstimulated' group). Neurons exhibiting Fos-like immunoreactivity were abundant at the stimulation site which included the infralimbic area, and dorsal peduncular cortex. Medullary Fos-like immunoreactivity observed in the 'Stimulated' and 'Unstimulated' groups exceeded levels observed in untreated rats and was detected in the rostral, caudal and intermediate areas of the ventrolateral medulla, and the commissural, intermediate, medial and lateral regions of the solitary tract nucleus, as well as the medial vestibular nucleus, and the dorsal motor nucleus of the vagus. The number of neurons displaying Fos-like immunoreactivity in the ipsilateral solitary tract nucleus and caudal ventrolateral medulla of the 'Stimulated' group was found to be significantly elevated compared to the contralateral side (P<0.05), and the 'Unstimulated' group bilaterally. Inhibition of solitary tract nucleus neurons using bilateral injections of the GABA(A) receptor agonist muscimol (44 pmol/25 nl) inhibited the sympathetic vasomotor baroreflex and attenuated the depressor and sympathoinhibitory response to medial prefrontal cortex stimulation by 62% and 65%, respectively. These findings suggest that the projection from the medial prefrontal cortex to the solitary tract nucleus is excitatory and support the hypothesis that the depressor response elicited by medial prefrontal cortex stimulation is mediated, in part, by a cortico-solitary projection which activates the intramedullary baroreflex pathway. PMID- 10362319 TI - Electrophysiological and morphological characterization of cytochemically-defined neurons in the caudal nucleus of tractus solitarius of the rat. AB - Morphological and electrophysiological properties of calbindin D-28k-, GABA- and dopamine-beta-hydroxylase-immunopositive neurons were investigated in the caudal nucleus of tractus solitarius of rats, using a patch-clamp whole-cell recording combined with intracellular staining and immunocytochemistry. Calbindin D-28K- and GABA-positive neurons had a small cell body (10.9+/-0.3 microm in diameter) and were distributed throughout the caudal nucleus of tractus solitarius. Double fluorescence immunocytochemistry revealed that calbindin- and GABA-positive neurons formed distinct subpopulations. Calbindin- and GABA-positive neurons double stained for biocytin showed extensive axon collaterals within the nucleus of tractus solitarius and some calbindin-positive, but not GABA-positive neurons, had also projection axons leaving the nucleus of tractus solitarius. Dopamine beta-hydroxylase-immunopositive neurons had a small (10.8+/-0.3 microm) or large (17.2+/-0.4 microm) cell body. Neurons with a small cell body were observed in the dorsomedial nucleus at the level of the area postrema, and in the area postrema, while neurons with a large cell body were observed in the medial nucleus throughout the caudal nucleus of tractus solitarius. Double fluorescence immunocytochemistry revealed that almost all small dopamine-beta-hydroxylase positive neurons were also immunoreactive for calbindin, while large dopamine beta-hydroxylase-positive neurons were not. Double staining for dopamine-beta hydroxylase and biocytin showed that neurons with a small cell body had moderate axon collaterals. On the contrary, neurons with a large cell body had few, if any, axon collaterals and a projection axon which could leave the nucleus of tractus solitarius. Following stimulation of the tractus solitarius, all neurons with a small cell body exhibited a polysynaptic excitatory response (type I neurons), while dopamine-beta-hydroxylase-immunopositive neurons with a large cell body exhibited a monosynaptic excitatory response (type II neurons) or an excitatory followed by an inhibitory response (type III neurons). Spontaneous and evoked excitatory postsynaptic currents of (type I neurons) calbindin- or GABA positive neurons were reversibly blocked by 6-cyano-7-nitroquinoxaline-2,3-dione. Spontaneous and evoked inhibitory postsynaptic currents of type III neurons were reversibly blocked by bicuculline. Type II neurons showed no spontaneous excitatory nor inhibitory postsynaptic currents. It was concluded that the three kinds of chemically-defined neurons formed distinct neuronal subpopulations in the caudal nucleus of tractus solitarius in terms of synaptic responses and morphological characteristics such as cell size and axonal trajectory. PMID- 10362320 TI - Detection of hypoxic cells with the 2-nitroimidazole, EF5, correlates with early redox changes in rat brain after perinatal hypoxia-ischemia. AB - The hypoxia-dependent activation of nitroheterocyclic drugs by cellular nitroreductases leads to the formation of intracellular adducts between the drugs and cellular macromolecules. Because this covalent binding is maximal in the absence of oxygen, detection of bound adducts provides an assay for estimating the degree of cellular hypoxia in vivo. Using a pentafluorintated derivative of etanidazole called EF5, we studied the distribution of EF5 adducts in seven-day old rats subjected to different treatments which decrease the level of oxygen in the brain. EF5 solution was administered intraperitoneally 30 min prior to each treatment. The effect of acute and chronic hypoxia on EF5 adduct formation (binding) was studied in the brain of newborn rats exposed to global hypoxia (8% O2 for 30, 90 or 150 min) and in the brain of chronically hypoxic rat pups with congenital cardiac defects (Wistar Kyoto). The effect of combined hypoxia ischemia was investigated in rat pups subjected to right carotid coagulation and concurrent exposure to 8% O2 for 30, 90 or 150 min. Brains were frozen immediately at the end of each treatment. Using a Cy3-conjugated monoclonal mouse antibody (ELK3-51) raised against EF5 adducts, hypoxic cells within brain regions were visualized by fluorescence immunocytochemistry. Brains from controls or vehicle-injected animals showed no EF5 binding. Notably, brains from animals which were chronically hypoxemic as a result of congenital cardiac defects also showed no EF5 binding. A short exposure (30 min) to hypoxia or to combined hypoxia-ischemia resulted in increased background stain and few scattered cells with low-intensity immunostaining. Acute hypoxia exposure of at least 90-150 min, which in this age animal does not result in frank cellular damage, produced patchy areas of low- to moderate-intensity fluorescence scattered throughout the brain. In contrast, 90-150 min of hypoxia-ischemia was associated with intense immunofluorescence in the hemisphere ipsilateral to the carotid occlusion, with a pattern similar to that reported previously for the histological damage seen in this model. This study provides a sensitive method for the evaluation of the level of oxygen depletion in brain tissue after neonatal hypoxia-ischemia at times much earlier than any method demonstrates apoptotic or necrotic cell death Since the level of in vivo formation of macromolecular adducts of EF5 depends on the degree of oxygen depletion in a tissue, intracellular EF5 binding may serve as a useful marker of regional cellular vulnerability and redox state after brain injury resulting from hypoxia-ischemia. PMID- 10362321 TI - Heterogeneity of the microglial response in photochemically induced focal ischemia of the rat cerebral cortex. AB - This study examined microglial responses after photochemically induced focal ischemia of the rat cortex. Microglial activation exceeded by far the area of the ischemic lesion. Based on morphological criteria and expression of immunomolecules three distinct patterns could be distinguished. (1) In the infarct core and the border zone microglia transformed into phagocytes and removed debris with the aid of hematogeneous macrophages. Exclusively in this area a subpopulation of CD8+ microglia/mnacrophages was present. (2) In secondarily degenerating fibre tracts and nuclei with retrograde neuronal loss, microglia were activated with a delay of days and showed increased expression of complement receptor 3, major histocompatibility complex class II and CD4 molecules, but only low phagocytic activity. (3) In remote ipsilateral cortex devoid of neuronal damage, microglia transiently responded by increased complement receptor 3, but not by major histocompatibility complex class II and CD4 expression. Furthermore, the total number of microglia had increased. This remote response could partly be blocked by dizocilpine maleate, a non-competitive N-methyl-D-aspartate receptor antagonist, implicating a functional role of spreading depression. Taken together, our findings point to a tight and differential regulation of microglial responses in the infarct core, degenerating fibre tracts and remote brain regions without neuronal loss. PMID- 10362323 TI - Chloride permeation across the Deiters' neuron plasma membrane: activation by GABA on the membrane cytoplasmic side. AB - Single plasma membranes were microdissected from Deiters' neurons freshly obtained from the lateral vestibular nucleus of the rabbit and their chloride permeability was studied in a microchamber system. The basal in-->out 36Cl- permeation initially found was brought to zero by Zn2+, 4,4' diisothiocyanatostilbene-2,2'-disulphonic acid and iodide. GABA on the membrane cytoplasmic side resulted in a measurable in-->out 36Cl- passage, which was blocked by the GABA(A) antagonists bicuculline and picrotoxin. This effect peaked at 1 microM GABA on the inner side of the membrane. At higher GABA concentrations, a strong desensitization of the effect was found. Stimulation of Cl- permeability by GABA on the extracellular side of the membrane peaked at much higher GABA concentrations, 10-100 microM. This excludes an effect due to passage of the neurotransmitter from the inner to the outer compartment in our microchamber device. Moreover, this possibility is also dismissed by the fact that 1 microM GABA on the membrane outside did not evoke any 36Cl- in-->out permeation. In addition, pentobarbitone by itself could also stimulate 36Cl- in- >out permeation when added on the cytoplasmic side of Deiters' membrane. On these bases and in agreement with our previous reports, we propose that structures behaving pharmacologically as GABA(A) receptors respond to low levels of GABA on the cytoplasmic side of these neurons' membranes. We suggest that these structures are devices that, at the expense of ATP consumed in their phosphorylation, extrude Cl- after postsynaptic GABA uptake into the Deiters' neuron. PMID- 10362322 TI - Induction of potassium channels in mouse brain microglia: cells acquire responsiveness to pneumococcal cell wall components during late development. AB - Lipopolysaccharides derived from cell walls of Gram-negative bacteria have proven a useful tool to simulate bacterial infection of the central nervous system. Rapid activation of microglia within the brain parenchyma as well as in vitro has thereby been shown to be an early event upon bacterial or lipopolysaccharide challenges. Less is known about microglial responses to a contact with Gram positive bacteria, such as Streptococcus pneumoniae, a lethal pathogen causing meningitis with a 30% mortality rate. In the present study, we compared lipopolysaccharide-induced microglial activation in vitro with that induced by preparations of pneumococcal cell walls. As a readout of microglial activation, we studied by patch-clamp recording the expression of outward rectifying potassium currents (IK+OR), which are known to be induced by lipopolysaccharide. We found that pneumococcal cell walls and lipopolysaccharide induced a similar type of IK+OR. Stimulation of IK+OR by pneumococcal cell walls and lipopolysaccharide involved protein synthesis since it was not induced in the presence of cycloheximide. Pharmacological characterization of the pneumococcal cell wall- and lipopolysaccharide-induced currents with specific ion channel blockers indicated for both cases expression of the charybdotoxin/margatoxin sensitive Kv1.3 subtype of the Shaker family of voltage-dependent potassium channels. Activation of the outward currents by pneumococcal cell walls depended on the developmental stage: while lipopolysaccharide triggered IK+OR in both embryonal and postnatal microglial cells, pneumococcal cell walls had only a marginal effect on embryonal cells. This, however, does not imply that embryonic microglial cells are unresponsive to pneumococcal cell walls. In both embryonic and postnatal cells, (i) the amplitude of the constitutively expressed inward rectifying potassium current was significantly reduced, (ii) tumor necrosis factor-a was released and (iii) the cells changed their morphology, similarly as it was induced by lipopolysaccharide treatment. Thus, embryonic microglial cells are sensitive to pneumococcal cell wall challenges, but respond with a distinctly different pattern of physiological reactions. The expression of IK+OR could thus be a suitable tool to study signalling cascades selectively involved in the activation of microglia by Gram-negative and -positive cell wall components and to functionally distinguish between populations of microglial cells. PMID- 10362324 TI - Long-term outcome after postmastectomy radiation therapy for the treatment of ductal carcinoma in situ of the breast. AB - Postmastectomy radiation therapy may be recommended for patients with a high risk for local recurrence after mastectomy for ductal carcinoma in situ (DCIS). However, long-term outcomes after postmastectomy radiation therapy are not well described. This study was performed to determine long-term outcomes in patients treated with radiation therapy after mastectomy for DCIS. The authors reviewed the records of all patients with breast cancer treated with postmastectomy radiation therapy between 1978 and 1992. Of 287 total patients treated, three (1%) were for DCIS. These three patients had diffuse microcalcifications on screening mammography. The reason for postmastectomy radiation therapy was a potentially increased risk for local recurrence because of a positive resection margin after mastectomy for DCIS. Surgery consisted of a total mastectomy (n = 2) or a modified radical mastectomy (n = 1). Radiation therapy consisted of 4275 5000 cGy to the chest wall in 200-225 cGy fractions. The energy used was 6-MV photons (n = 2) or 15-MV photons (n = 1). No regional nodal irradiation was used. Bolus was applied to the chest wall every other day in one of the three patients. One patient was treated with a scar boost after chest wall irradiation (boost dose, 1000 cGy; total dose, 5275 cGy). The median age for the three patients was 46 years (range, 41-68 years). No patient received adjuvant chemotherapy or hormonal therapy. With a minimum follow-up of 7.1 years (median, 7.4 years; range, 7.1-19.4 years), no local-regional recurrence or evidence of metastatic disease developed in any of the patients. No long-term complication from radiation therapy was noted, and no contralateral breast cancer developed. All patients were alive and free of relapse at the last follow-up. The use of radiation therapy in this group of three patients has shown no evidence of relapse with a minimum of 7.1 years of follow-up. The authors conclude that radiation therapy may be indicated after mastectomy for DCIS to reduce the risk of recurrence for high-risk patients. PMID- 10362325 TI - Phase II trial of topotecan in advanced breast cancer: a Cancer and Leukemia Group B study. AB - The topoisomerase I inhibitor topotecan had demonstrated good antitumor activity in several murine tumor systems and in human clonogenic assays by 1993. In that year, the Cancer and Leukemia Group B (CALGB) began a phase II trial to determine its activity in patients with breast cancer who had previously received one course of chemotherapy for advanced breast cancer. Between April 1993 and June 1994, 53 patients of performance status 0-2 entered the study, of whom 47 were eligible and 40 were evaluable. Topotecan was given at a dose of 1.5 mg/m2 over 30 minutes daily for 5 days every 21 days. In the absence of progression or withdrawal of consent, therapy was continued indefinitely. The median age was 58 years (range 30-79). There were no complete responses and four partial responses, resulting in an objective response rate of 10% (95% CI: 3-24%). Responses were noted in lymph nodes, liver, and skin. The median duration of response was 5 months. The median survival was 12 months. Life-threatening toxicities were almost exclusively hematologic. However, myelosuppression was not cumulative. It was concluded that topotecan has only modest activity among women with advanced breast cancer who have previously received one course of chemotherapy. Given its modest activity and predominant hematologic toxicity, it does not appear to be a promising drug for either single-agent or combination chemotherapy in the salvage setting of advanced breast cancer. PMID- 10362326 TI - Renal cell carcinoma: a paradigm of lanthanic disease. AB - Renal cell carcinoma is characterized by varied and sometimes obscure manifestations, which include unusual metastatic sites and paraneoplastic and vascular syndromes. In this review, uncommon metastatic sites and their clinical significance are discussed, particularly the thyroid, nasal structures, vagina, and gastrointestinal sites. Paraneoplastic syndromes appear to be related predominantly to cytokines or immunologic mechanisms. Vascular syndromes are related to the tendency of the tumor to spread by direct venous extension and to complications related to the vascularity of the tumor or its metastases. The recognition of unusual manifestations of renal cell carcinoma is important because these syndromes may lead to the diagnosis. Moreover, paraneoplastic syndromes and vascular findings may not indicate unresectability or incurability. PMID- 10362327 TI - Bulky, barrel-shaped cervical carcinoma (stages IB, IIA, IIB): the prognostic factors for pelvic control and treatment outcome. AB - The purpose of this study was to assess the prognostic factors for pelvic control and the treatment outcome in bulky, barrel-shaped cervical carcinomas. Between September 1980 and December 1992, 65 patients with stage IB or stage IIA-B carcinoma of the uterine cervix classified as barrel-shaped or concentrically expanded (i.e., at least 5 cm in greatest diameter) were treated with curative intent. Forty patients had stage IB or stage IIA carcinoma (according to the classification of the International Federation of Gynecology and Obstetrics [FIGO]), and 25 patients had FIGO stage IIB carcinoma. Seventy-two percent of the patients were treated with radiotherapy (RT) alone and 28% with radiotherapy followed by extrafascial hysterectomy (RT + S). The median follow-up time of surviving patients was 68 months (range 33-172). Survival and control rates were calculated by the Kaplan-Meier method. The 10-year actuarial pelvic control rate was 75% for all patients. The likelihood of pelvic control was not affected by FIGO stage, tumor size, patient's age, histologic features, or treatment modality (RT vs. RT + S). The extent of tumor regression following external beam radiotherapy correlated with the likelihood of local control (p = 0.02). For patients treated with RT alone, increased brachytherapy dose was associated with an increased likelihood of local control. The 10-year actuarial overall and cause specific survival rates were 53% and 68%, respectively, and did not differ significantly between treatment groups. It is concluded that for most patients with bulky cervical carcinoma, RT alone provides good local control and survival. However, for patients with tumors that respond poorly to external beam radiotherapy, local control and survival are poor. More aggressive treatment protocols should be considered for these patients. The routine use of adjuvant hysterectomy is not recommended. PMID- 10362328 TI - Mature results from a phase II trial of accelerated induction chemoradiotherapy and surgery for poor prognosis stage III non-small-cell lung cancer. AB - Mature results are reported from a phase II trial of accelerated induction chemoradiotherapy and surgical resection for stage III non-small-cell lung cancer whose prognosis is poor. Surgically staged patients with poor prognosis stage III non-small-cell lung cancer were eligible for this study. Four-day continuous intravenous infusions of cisplatin 20 mg/m2/day, 5-fluorouracil 1,000 mg/m2/day, and etoposide 75 mg/m2/day were given concurrently with accelerated fractionation radiation therapy, 1.5 Gy twice a day, to a total dose of 27 Gy. Surgical resection followed in 4 weeks. Identical postoperative chemotherapy and concurrent radiation to a total dose of 40 to 63 Gy was subsequently given. Between February 1991 and June 1994, 42 eligible and evaluable patients, 23 with stage IIIA disease and 19 with stage IIIB disease, were entered in this trial. Treatment was well tolerated. The pathologic response rate was 40%. This response was complete in 5%. With a median follow-up of 54 months, the Kaplan-Meier 4-year survival estimate is 19%: 26% for stage IIIA and 11% for stage IIIB patients. Patients with a pathologic response, resectable disease, or pathologic downstaging to stage 0, I, or II had a better survival. The 4-year estimates of locoregional and distant disease control are 70% and 19%, respectively. It is concluded that although the ultimate role of concurrent chemoradiotherapy and surgery in stage III non-small-cell lung cancer must await the results of phase III clinical trials, survival and locoregional control in this study appear improved in comparison with historical experience. There is a subset of patients, able to undergo resection with pathologic downstaging, who have a projected survival equivalent to that of patients with more limited disease. Clinical or pathologic tools to identify these patients before treatment would be highly useful. PMID- 10362329 TI - Diffuse malignant leptomeningeal gliomatosis in a child: a case report and review of the literature. AB - Diffuse leptomeningeal gliomatosis is a rare condition characterized by glioma in the leptomeninges without a dominant mass lesion. The difficulty in diagnosis of this condition, its rarity. and its extensive nature have hampered its successful treatment. Most cases of primary diffuse leptomeningeal gliomatosis have occurred in adults. Reported here is a case of this condition in a 9-year-old girl; to the authors' knowledge, she is the youngest patient with diffuse leptomeningeal gliomatosis and the longest survivor of the malignant variety. PMID- 10362330 TI - Marimastat in patients with advanced pancreatic cancer: a dose-finding study. AB - Patients with solid tumors, including carcinoma of the pancreas, express high levels of matrix metalloproteinases (MMP), and these enzymes are believed to be important for the growth, spread, and dissemination of most solid malignant tumors. Marimastat is the first orally available MMP inhibitor (MMPI) to be tested in humans and has been shown to inhibit the spread and growth of pancreatic cancer in animal models. The purpose of the present study was to define the toxicities, safety, and tolerance of various doses of marimastat and also to get an early indication of potential biologic activity in patients with advanced pancreatic cancer. The authors prospectively studied 64 patients with advanced carcinoma of the pancreas in whom standard treatments had failed. Eligible patients had a progressive rise in CA 19/9 levels of >25% over the 4 week period preceding their entry into the study. Patients were studied in groups of 8 to 10, with each group receiving escalating dosages ranging from 5 mg twice daily to 75 mg twice daily and 10 to 25 mg daily. Patients were considered for long-term (beyond 4 weeks) continuation treatment if clinical benefit, in the view of the investigator, was derived. Study endpoints were safety, tolerance, and changes in the rate of rise of CA 19/9, which were used as surrogate markers for disease progression. Marimastat was well tolerated. Musculoskeletal pain, stiffness, and tenderness emerged as dose-limiting toxicity. No other dose related toxicities were observed. A reduced rate of rise of CA 19/9 was observed at dose levels of 5, 10, and 25 mg twice daily. The overall median survival was 160 days, with a 1-year survival of 21%. Marimastat was associated with an acceptable toxicity profile, and these preliminary data suggest that long-term oral administration is feasible and safe. Doses of 5, 10, and 25 mg twice daily were identified as the optimal doses to be tested in larger randomized studies. PMID- 10362331 TI - Adjuvant (cisplatin, etoposide, and 5-fluorouracil) chemotherapy after curative resection of gastric adenocarcinomas involving the esophagogastric junction. AB - Gastric adenocarcinomas involving the esophagogastric junction represent a particular therapeutic problem because they lie in the border area between two body cavities: the thorax and the abdomen. The prognosis of gastric adenocarcinomas involving esophagogastric junction is poor because there is widespread lymphatic metastasis, making curative resection difficult. Even in patients with localized disease who are potentially curable, the 5-year survival rate is approximately 20% with curative resection only, somewhat lower than for those with cancer elsewhere in the stomach. The authors conducted a pilot study to evaluate the safety and possible efficacy of adjuvant chemotherapy with cisplatin, etoposide, and 5-fluorouracil (PEF) after curative resection of gastric adenocarcinoma involving esophagogastric junction. Three cycles of adjuvant PEF chemotherapy with cisplatin (20 mg/m2/day intravenously on days 1 5), etoposide (100 mg/m2/day intravenously on days 1, 3, and 5), and 5 fluorouracil (800 mg/m2/day continuous intravenous infusion on days 1-5) were given every 3 weeks after curative resection of gastric adenocarcinoma involving the esophagogastric junction. Between November 1989 and June 1995, a total of 50 patients with postoperative stage II, IIIA, or IIIB disease entered this trial. In 14 of 50 patients (28%), the disease recurred during the follow-up of 4-83 months (median 26 months). Disease-free survival was 4-83+ months (median 48 months), and the actuarial 5-year disease-free survival rate was 48% (95% CI: 41% to 55%). Overall survival was 4-83+ months (median 62 months), and the actuarial 5-year survival rate was 54% (95% CI: 40% to 68%). The prognostic factor analysis showed that the postoperative N stage and the interval between surgery and chemotherapy affected disease-free survival and overall survival. The toxicities of PEF adjuvant chemotherapy were leukopenia, nausea/vomiting, and alopecia, but they were mostly mild and reversible except in one patient who died because of treatment-related sepsis. Adjuvant chemotherapy with three cycles of PEF regimen was well tolerated and seems to be a promising treatment for gastric adenocarcinoma involving the esophagopstric junction, in comparison with previous treatments. To define the efficacy of adjuvant PEF chemotherapy for gastric adenocarcinoma involving esophagogastric junction, prospective randomized trials are warranted. PMID- 10362332 TI - Phase III controlled evaluation of glutamine for decreasing stomatitis in patients receiving fluorouracil (5-FU)-based chemotherapy. AB - Mucositis is a prominent dose-limiting toxicity associated with 5-FU-based chemotherapy. On the basis of preliminary data suggesting that the amino acid glutamine could alleviate this problem, the authors developed this trial. Patients scheduled to receive their first 5-FU-based chemotherapy regimen were selected for study. Following stratification, patients were randomized, in a double-blind manner, to receive oral glutamine or a placebo preparation in a prophylactic manner. Patients in both groups were given oral cryotherapy before chemotherapy and were evaluated for mucositis by standard physicians' evaluation and by a self-report instrument. Sixty-six patients were randomized to receive glutamine and 68 to receive the placebo preparation. There were no significant differences or substantial trends in the mucositis scores between the two study arms as measured by either the physicians or the patients. It was concluded that the dose and schedule of glutamine used in this clinical trial does not alleviate 5-FU-induced mucositis. PMID- 10362333 TI - Epirubicin, folinic acid, fluorouracil, and etoposide in the treatment of advanced gastric cancer: phase II study of the Southern Italy Oncology Group (GOIM). AB - In the authors' previous experience, the addition of epidoxorubicin to the FA-FU regimen obtained a better response rate than that of FA-FU alone in patients with advanced gastric cancer. Furthermore, considering the good efficacy and mild toxicity observed with the addition of etoposide to the FA-FU combination in the German study, the authors conducted a trial to explore the efficacy and tolerability of the ELFE regimen (epirubicin, folinic acid, fluorouracil, and etoposide) in previously untreated advanced gastric cancer patients. Of the 55 patients entered, 51 were evaluable for efficacy. Four complete responses (8%) and 21 partial responses (41%) were observed, with an overall response rate of 49% (95% CI: 35-63%). The median duration of response and survival were 6 and 8 months, respectively. Responder patients showed a significantly better median survival duration than nonresponders (12 vs. 4 months, respectively; p < 0.0001). Toxicity was evaluated in all patients: only 1 patient refused to continue therapy despite low toxicity. As expected, the major toxicities observed were gastrointestinal disturbance, leukopenia, and loss of hair. In conclusion, the ELFE combination regimen appears to be effective and well tolerated for the treatment of advanced gastric cancer patients. PMID- 10362334 TI - Phase I-II trial of intensification of the MAID regimen with support of lenograstim (rHuG-CSF) in patients with advanced soft-tissue sarcoma (STS). AB - This study was conducted to determine the maximum tolerated dose of an intensified MAID (mesna, adriamycin, ifosfamide, dacarbazine) regimen with the support of lenograstim in patients with advanced soft tissue sarcomas. Following 1 cycle of MAID at the standard dose, four patients were to be treated at each of five dosage levels: +25%, +45%, +65%, +85%, +100%. Sixteen patients were treated. Because there were no significant differences in hematologic toxicity between patients receiving lenograstim 5 or 10 microg/kg/day (levels 1-5 and 1-10), the data were pooled for comparison with level 2. The median duration of absolute neutrophil count < 0.5 x 10(9)/l was 3 days at level 1 and 7 days at level 2 (p < 0.01). The median platelet nadir was 25 x 10(9)/l at level 1 and 10 x 10(9)/l at level 2 (p < 0.01). The median duration of toxicity-related hospitalization was 3.5 days and 11 days at levels 1 and 2, respectively, (p < 0.001). Mucositis > or = grade III occurred after 3/29 cycles at level 1 and 10/15 cycles at level 2 (p < 0.001). After 4 cycles at level 1, 8/8 patients still had performance status scores < or = 2, and only 4/8 had performance status scores < or = 2 after the second cycle at level 2. Lenograstim enabled an increase of 25% of the MAID regimen. At higher dose levels, severe mucositis and deterioration in performance status were dose limiting. PMID- 10362335 TI - First-line treatment with mitoxantrone, methotrexate, vincristine, and carboplatine (MIMOC) plus cyclical hormonotherapy with tamoxifen and megestrol acetate in advanced breast cancer. AB - Fifty patients with stage IIIB and IV breast cancer entered a prospective study receiving combination chemotherapy consisting of mitoxantrone (8 mg/m2) on day 1, methotrexate (30 mg/m2) on day 1, vincristine (1 mg/m2) on day 2, and carboplatine (250 mg/m2) on day 2 (MIMOC), plus cyclical hormonotherapy with tamoxifen (20 mg daily, days 1-10) and megestrol acetate (160 mg daily, days 11 21). The regimen was repeated every 3 weeks. None had received chemotherapy for advanced disease, although 17 patients had previously received adjuvant chemotherapy and 21 had received adjuvant hormonotherapy with tamoxifen. Twenty seven patients had positive estrogen receptor (ER+) status, and 23 negative estrogen receptor (ER-) status. Response was observed in 31 (62%) of the 50 analyzed patients (95% CI: 48.5-75.4%), with 5 complete responses (10%). A significantly better response rate was observed in ER+ patients (p = 0.03). The median duration of response was 16 months, and the median time to disease progression was 18 months. The median overall survival was 19 months (27 for responders and 7 for nonresponders). ER+ patients had a higher probability of survival (p = 0.02). Toxicity was moderate. Nausea/vomiting and myelotoxicity were the main side effects. In conclusion, MIMOC plus cyclical hormonotherapy represents a well-tolerated and effective first-line treatment for advanced breast cancer. The observed difference in response and survival in favor of ER+ patients warrants further investigation. PMID- 10362336 TI - Variations of p53 in cultured fibroblasts of patients with lung cancer who have a presumed genetic predisposition. AB - The authors studied cultured skin fibroblasts of 64 patients with lung cancer for constitutive mutations of the p53 tumor suppressor gene by using polymerase chain reaction and single-strand conformation polymorphism covering the entire coding region. The patients were considered to be genetically predisposed because lung cancer had developed in 25 of them before age 46 and because 42 of them had at least one first-degree relative with lung cancer. One mutation was detected at position 235 coding for serine instead of asparagine in the conserved DNA binding domain. The Pro/Pro genotype at codon 72 of p53, considered to harbor an increased risk for lung cancer, could not be detected with increased frequency in this study's patients. From these data, the authors conclude that constitutive variations of the p53 gene do not represent a major genetic determinant for lung cancer. PMID- 10362337 TI - Non-Hodgkin lymphoma and coexisting primary cancers: a retrospective clinical analysis of 10 patients. AB - The simultaneous occurrence of non-Hodgkin lymphoma (NHL) and primary cancers is rare, and the treatment strategy for both malignancies is unclear. The authors analyzed the clinical records of 10 patients with NHL and coexisting primary cancers. All patients initially had symptoms of NHL, and all carcinomas were found at the initial workup of NHL by chance. The most common primary sites of coexisting cancers were the stomach (six patients) and the colon (two). Histologically, the majority of NHLs were intermediate grade, and all lesions were B-cell type. All primary cancers were adenocarcinoma. Initially, NHL was treated with radiotherapy or chemotherapy. Six primary cancers were resected surgically or endoscopically after the remission of NHL. The remaining four patients received no treatment for primary cancers because of advanced stages or early relapse of NHL. Three patients died of NHL, one died of cancer, and six were still alive, five without evidence of disease and one with disease. The authors conclude that early detection of a coexisting cancer and appropriate treatment after the remission of NHL may increase the possibility of a cure. PMID- 10362338 TI - Palliative treatment of epidemic Kaposi sarcoma of the feet. AB - Limited information is available in the medical literature on epidemic Kaposi sarcoma (EKS) of the foot. Patients with EKS of the foot can experience severe discomfort that makes it difficult to ambulate and even wear shoes. Between 1985 and 1996, 36 patients with EKS of the foot were treated with palliative intent. Most patients were referred for radiation therapy because of foot discomfort or marked difficulty with ambulation. From the pool of 36 patients, data were available at completion of treatment for 46 sites, and at 1 month for 44 sites. Morbidity was assessed for 35 sites. The median follow-up time for the 44 sites with at least 1 month follow-up was 8 months. The most frequently used regimen was a novel fractionation schedule of three fractions a week at 3.5 Gy/fx to a total dose of 21.0 Gy. The overall response rate and complete response rate for the 44 sites with at least 1 month follow-up were 91% and 80%, respectively. The 46 treated sites evaluated at the completion of treatment had a complete response rate of only 13% and an overall response rate of 63%. Of the 35 sites assessed for acute toxicity, 63% experienced discomfort related to the radiation therapy. This discomfort usually resolved without intervention within 2 weeks of completion of radiation therapy. For patients with and without a history of opportunistic infections, complete responses were observed in 8 of 12 sites (67%) and 25 of 27 sites (93%), respectively (p = 0.06). Radiation therapy for EKS of the foot yields excellent response rates, comparable with responses seen in other cutaneous sites with EKS. Appropriate patient education and support are needed because initial responses to radiation therapy are often disappointing and pedal discomfort can be exacerbated transiently. However, the discomfort resolves and complete response occurs in most patients. The 3.5-Gy triweekly fractionation schedule is a convenient and effective regimen and minimizes treatment visits for patients with ambulatory discomfort. A history of opportunistic infections appears to be a poor prognosticator of response to radiation treatments. PMID- 10362339 TI - Phase II evaluation of treatment of complete resection of hepatic metastases from colorectal cancer and adjuvant hepatic arterial infusion of floxuridine: an Eastern Cooperative Oncology Group Study (PB083). AB - This study was designed to evaluate hepatic arterial infusion of floxuridine (FUDR) in patients with resected hepatic metastases from colorectal cancer. Patients who met eligibility criteria had an Infusaid pump (Infusaid Corporation, Sharon, MA, U.S.A.) implanted for intraarterial administration of chemotherapy. After complete surgical resection of hepatic metastases, FUDR (0.2 mg/kg/day) was given in 28-day cycles consisting of 14 days of treatment followed by 14 days of rest. Of 11 patients enrolled, one was ineligible, one received no treatment because of a blocked pump, and nine were treated per protocol. Of the nine treated patients, all are dead: one from hepatic toxicity, one from unrelated causes, and seven from progressive disease. Grade 3-4 toxicity included three cases of gastritis and two cases of hepatotoxicity from FUDR. Although this regimen was not successful, in part because of toxicity, the patient population studied here should be considered for future studies of adjuvant therapy. PMID- 10362340 TI - Short-term intensive consolidation therapy after all-trans retinoic acid in acute promyelocytic leukemia. AB - Complete remission induced by all-trans retinoic acid (ATRA) in acute promyelocytic leukemia is short lived, and several consolidation chemotherapy courses usually are given to reduce the relapse rate. To assess the value of short-term intensive consolidation, 38 patients with newly diagnosed acute promyelocytic leukemia entered a prospective study in which induction therapy with ATRA immediately was followed by a single course of mitoxantrone plus high dose cytarabine (3 g/m2 every 12 hours, days 1-4), with no further treatment. Complete remission was achieved in 31 patients (81.6%) after a median time of 49 days of ATRA (to which chemotherapy was added at entry in 10 patients with leukocytosis). Thirty patients received the planned consolidation course. After a median follow-up of 36 months, four of these patients have relapsed and 24 are still in first complete remission, for an estimated disease-free survival of 75% at 60 months. The authors conclude that this single course consolidation of ATRA induced remission provides excellent long-term control of acute promyelocytic leukemia. PMID- 10362341 TI - Combination cisplatin-vinorelbine for relapsed and chemotherapy-pretreated metastatic breast cancer. AB - The purpose of this study was to evaluate the combination of cisplatin and vinorelbine (PVn) for relapsed and chemotherapy-pretreated metastatic breast cancer. Twenty-three patients with metastatic breast cancer and prior chemotherapy were entered in a phase II study between June 1993 and December 1994. Eleven patients were premenopausal and 12 were postmenopausal. Follow-up data up to June 1997 are presented. All patients received cisplatin at a dose of 90 mg/m2 divided over 3 days as 30 mg/m2 infused over 4 hours. Intravenous vinorelbine 25 mg/m2 was given on days 1 and 8 or 15 according to patients' blood counts. Cycles were given every 3 to 4 weeks. An overall response rate of 61% (16/23 patients) was observed. Complete remission was obtained in six patients (26%) and partial remission was obtained in nine patients (35%). The duration of response ranged from 3 to 9 months, with an average of 4 months. Stable disease was noted in 29.1% and progressive disease in 8.3%. Overall survival at 12 months was 50%, and at 36 months it was 8%. Five of 12 patients (42%) who had prior doxorubicin therapy responded well to cisplatin-vinorelbine. Of those 12, seven were refractory and progressive on a doxorubicin-containing regimen, one had complete remission, and four had partial remission. Hematologic toxicity was acceptable. Treatment was delayed because of neutropenia in nine cycles (9.2%) and grade 2 leukopenia occurred in 54% of cycles. Febrile neutropenia occurred in seven cycles (7.1%), and five cycles were complicated by documented sepsis (5.1%). No treatment-related mortality occurred. Thrombocytopenia (grade 3) was seen in 27% of cycles, with no patient having a platelet count below 50,000 or bleeding episodes. Other toxicities were not major or dose-limiting. In conclusion, the combination of cisplatin and vinorelbine produced good responses: 61% response rate (16 of 23 patients) in relapsed, refractory, and heavily pretreated metastatic breast cancer, with 50% survival at 1 year, 12% at 2 years, and 8% at 3 years. In addition, a response rate of 42% (5 of 12 patients) was seen in patients resistant to anthracyclines. PMID- 10362342 TI - Bone marrow relapse in primary mucinous carcinoma of skin. AB - Primary mucinous carcinoma of skin is a rare adnexal tumor arising from the eccrine sweat gland. The tumors grow slowly and have low rates of local recurrence and rare chances of distant metastasis. The authors report a 70-year old man with primary mucinous skin carcinoma who had a relapse in bone marrow 19 months after initial treatment. PMID- 10362343 TI - Cisplatin ototoxicity: the importance of baseline audiometry. AB - The role and optimal use of audiometry in monitoring for cisplatin ototoxicity are incompletely defined. Audiograms were obtained from 217 patients before treatment with cisplatin-based chemotherapy for cancers of the esophagus, lung, or head and neck. Posttreatment audiometry then was conducted in 53 of these patients. Chemotherapy consisted of two (87%) or three (13%) courses of cisplatin at a dose of 20 mg/m2/day given as a continuous intravenous infusion over 4 days. Simultaneous 5-fluorouracil or paclitaxel also was given, and 38% received concurrent radiation therapy to the head and neck. Air-conduction thresholds for each ear were obtained at 250, 500, 1000, 2000, 4000, 6000, and 8000 Hz. Three three-frequency pure-tone averages (PTA) also were calculated. Framingham gender specific, age-adjusted norms were used, beginning at age 60 to correct for presbycusis, and the upper limit of normal was calculated as the greater of the Framingham mean plus twice the standard error, or 25 dB. Hearing abnormality was defined as a threshold >10 dB above the norm for any PTA, or >20 dB above the norm for any individual frequency. Hearing loss was defined as an elevation over baseline threshold of >10 dB for any PTA or >20 dB for any individual frequency. Of the 217 patients who underwent baseline testing, 57 (26%) were found to have hearing abnormality in excess of the expected presbycusis. Post-cisplatin audiograms demonstrated hearing loss in 19 of the 53 retested patients (36%) when compared with their own baseline. As determined by tympanometry, none of these subjects had a conductive component to their hearing loss. These observations were independent of the duration of follow-up after treatment and of the total dose of cisplatin administered. The authors conclude that significant preexisting hearing abnormality is common in this patient population and that, even after low dose cisplatin administration, additional hearing loss occurs frequently. Baseline testing is mandatory if follow-up studies are to be adequately interpreted. PMID- 10362344 TI - Combined chemoradiotherapy for unresectable pancreatic cancer. AB - This study was undertaken to evaluate the efficacy of a regimen of combined chemoradiotherapy in patients with unresectable adenocarcinoma of the pancreas. An analysis was undertaken on 27 patients from January 1992 to May 1996. Patients had a median age of 70 years (range, 40-78) and Eastern Cooperative Oncology Group Performance Status of 0-2. Eighteen patients had locoregional disease (T2 T3, N0-N1, M0), and nine had metastatic disease. Chemotherapy consisted of four cycles of 5-fluorouracil 1 gm/m2/day as a continuous infusion over 110 hours, streptozotocin 300 mg/m2/day over 30 minutes on days 2-4, and cisplatin 100 mg/m2 over 2 hours on day 4 only, followed by a maintenance regimen of 5-fluorouracil and leucovorin every 2 weeks. The radiotherapy was administered as a split course concurrently with chemotherapy to a total dose of 6000 cGy. Toxicity was frequent, but there were no treatment-related deaths. Grade III and IV toxicity was primarily limited to myelosuppression, stomatitis, and gastrointestinal side effects. Fifteen patients (56%) were able to complete either three or four cycles of chemoradiotherapy. All patients were evaluable for toxicity, response, and survival. Nine patients (33%) had an objective response (four complete response 5 partial response), two remained stable, and 16 (59%) had disease progression. Median survival for the entire group was 19 weeks (2-139), and the median survival for overall responders was 56 weeks (15-139). No patient with localized disease underwent subsequent surgical resection. The authors conclude that those patients who are able to tolerate the entire treatment regimen may achieve a useful prolongation of time to tumor progression. PMID- 10362345 TI - Extrahepatic regional chemotherapy: use of technetium-99m labeled macroaggregated albumin. AB - The purpose of this study was to verify the applicability of nuclear techniques with technetium-99m labeled macroaggregated albumin (Tc-99m-MAA) in extrahepatic regional chemotherapy. Of 98 patients in whom arterial Port-a-caths were implanted by transcutaneous access, 13 were treated by regional extrahepatic chemotherapy (breast, one; pancreas, four; kidney, one; uterus, three; vagina, two; bladder, two). In all 13 patients, Tc-99m-MAA was slowly infused intraarterially. The examination showed the perfusion of the area with the neoplasm and excluded the presence of important misperfusions of Tc-99m-MAA to the nearest areas. To detect the presence of an arteriovenous shunt with systemic misperfusion, an anterior image of the thorax was obtained in all patients and an index of misperfusion was calculated. In 12 patients, the index was < 5%; in one patient it was about 40%. In conclusion, our preliminary experience concerns the monitoring of intraarterial infusion chemotherapy of extrahepatic districts. In all 13 patients, we evaluated the correct positioning of the intraarterial catheter and the distribution pattern of the arterial flow, with a semiquantitative indication of arteriovenous shunting. This method gave us an instrument of study that was inexpensive, harmless, and free of collateral complications. PMID- 10362346 TI - Carcinoma of the stomach metastatic to the liver that progressed after hepatic arterial infusion of cisplatin plus 5-floxuridine, and then dramatically regressed after chemoembolization based on positive chromogranin staining. PMID- 10362347 TI - Cardiac glycosides in the next millennium. AB - Despite the documented efficacy of cardiac glycosides in improving symptoms in patients with heart failure caused by systolic ventricular dysfunction, considerable debate continues as to whether the use of this class of drugs should continue into the next millennium. In this review, the authors briefly examine the basic pharmacology of these drugs relevant to the treatment of heart failure, emphasizing their role in reducing sympathetic nervous system activity in patients with advanced heart failure. Next, withdrawal trials and the Digoxin Investigation Group dataset are reviewed in some detail. Despite these important additional data on the safety and efficacy of digitalis use in heart failure that became available in the 1990s, considerable controversy remains. Perhaps most importantly, if the mechanism by which these drugs improve symptoms in patients with heart failure is principally mediated by sympatholytic activity, do they remain relevant as beta-adrenergic antagonists become standard therapy for this disease? PMID- 10362348 TI - The role of organic nitrates in the treatment of heart failure. AB - Nitrates have been widely used in the treatment of patients with chronic congestive heart failure. Although the use of these drugs has not been approved by the Food and Drug Administration, multiple studies have shown their favorable effects. Organic nitrates have been shown to have a beneficial effect on ischemia, hemodynamic profile, magnitude of a mitral regurgitation, endothelial function, and cardiac remodeling. These drugs, when used in combination with hydralazine, have improved exercise capacity and survival. Recent studies have shown that the use of nitrates in patients already treated with standard heart failure therapy, including angiotensin converting enzyme (ACE) inhibitors, resulted in hemodynamic improvement, marked enhancement of exercise tolerance, reduction of left ventricular size, and augmentation of systolic function. These data suggest a role for organic nitrates as an adjunctive therapy to ACE inhibitors in patients with chronic heart failure and for nitrates in combination with hydralazine as an alternative treatment in patients who are intolerant to ACE inhibitors. PMID- 10362349 TI - Angiotensin-converting enzyme inhibitors and angiotensin II receptor antagonists in the treatment of heart failure caused by left ventricular systolic dysfunction. AB - Activation of the renin-angiotensin-aldosterone system (RAAS) in left ventricular systolic dysfunction is a critically important determinant in the pathophysiologic processes that lead to progression of heart failure and sudden death. Angiotensin II, acting at the specific angiotensin receptor (AT1-R), activates a series of intracellular signaling sequences which are ultimately expressed within the cardiovascular system as vasoconstriction and associated vascular hypertrophy and remodeling. Angiotensin converting enzyme (ACE) inhibition leads to increases in the vasodilatory peptides bradykinin and substance P and at least an initial reduction in angiotensin II concentrations. AT1-R blocking drugs prevent access of angiotensin II to the AT1-R and thus prevent cellular activation. ACE inhibitors have clearly been demonstrated through a large number of clinical trials to increase survival in congestive heart failure, primarily by reducing the rate of progression of left ventricular dilatation and decompensation. However, this beneficial effect diminishes over time. Preliminary short-term clinical studies evaluating the efficacy of AT1-R blocking drugs in the treatment of heart failure have suggested that they elicit similar hemodynamic and neuroendocrine effects as do the ACE inhibitors. The combination ACE inhibitors and AT1-R blocking drugs offer the theoretical advantage of increasing bradykinin while blocking the actions of angiotensin II, and thus possibly show a synergistic effect. Again, preliminary studies have yielded encouraging results that are difficult to interpret because neither ACE inhibitor nor the AT1-R blocking drug doses were titrated to tolerance. Pharmacological manipulation of the RAAS has led to better understanding of its role in heart failure and improved clinical outcomes. PMID- 10362350 TI - Beta blocker treatment in heart failure. AB - The sympathetic nervous system occupies a prominent role in heart failure both as a marker of severity of disease and also as an important factor in its progression. Beta blocker therapy, once thought heretical in heart failure, has consistently improved cardiac function and slowed progression of disease. Large clinical trials of mild to moderate heart failure show improved survival as well as reduction in hospitalization. Beta blockers now have stronger data in heart failure than converting enzyme inhibitors, and should be considered standard therapy in mild-moderate heart failure. Ongoing trials are addressing beta blocker therapy in advanced heart failure and comparisons between agents. PMID- 10362351 TI - Inhibition of E6 induced degradation of p53 is not sufficient for stabilization of p53 protein in cervical tumour derived cell lines. AB - The E6 proteins derived from tumour associated papillomavirus types target the cellular tumour suppressor protein p53 for ubiquitin mediated degradation. In cell lines derived from cervical tumours the p53 protein is present in very low amounts, but it can be activated by appropriate DNA damaging agents, indicating that functional p53 is present within these lines. Recent studies have also shown that different polymorphic forms of the p53 protein are differentially susceptible to E6 mediated degradation. Therefore we have been interested in analysing the effects of different HPV E6 proteins upon p53 levels in a variety of cervical tumour derived cell lines. We show that inhibition of E6 mediated degradation of p53 frequently results in increased levels of p53 expression. However, there are notable exceptions to this where increased p53 levels are only obtained following DNA damage and proteasome inhibition. We also show in E6 expressing cells, that as well as p53 being targeted for degradation, the localization of p53 to the nucleus is also inhibited, consistent with previous observations which indicate that degradation of p53 is not essential for E6 mediated inhibition of p53 function. These results have important implications for any potential therapies which might aim to block E6 mediated degradation of p53. PMID- 10362352 TI - The Bcl-3 oncoprotein acts as a bridging factor between NF-kappaB/Rel and nuclear co-regulators. AB - The proto-oncoprotein Bcl-3 is a member of the IkappaB family and is present predominantly in the nucleus. To gain insight into specific nuclear functions of Bcl-3 we have isolated proteins that interact with its ankyrin repeat domain. Using the yeast two-hybrid-system we identified four novel binding partners of Bcl-3 in addition to NF-kappaB p50 and p52, previously known to associate with Bcl-3. The novel Bcl-3 interactors Jab1, Pirin, Tip60 and Bard1 are nuclear proteins which also bind to other transcription factors including c-Jun, nuclear factor I (NFI), HIV-1 Tat or the tumor suppressor and PolII holoenzyme component Brca1, respectively. Bcl-3, p50, and either Bard1, Tip60 or Pirin are sequestered into quarternary complexes on NF-kappaB DNA binding sites, whereas Jab1 enhances p50-Bcl-3-DNA complex formation. Furthermore, the histone acetylase Tip60 enhances Bcl-3-p50 activated transcription through an NF-kappaB binding site, indicating that quarternary complexes containing Bcl-3 interactors modulate NF kappaB driven gene expression. These data implicate Bcl-3 as an adaptor between NF-kappaB p50/p52 and other transcription regulators and suggest that its gene activation function may at least in part be due to recruitment of the Tip60 histone actetylase. PMID- 10362354 TI - Regulation and function of protein kinase B and MAP kinase activation by the IL 5/GM-CSF/IL-3 receptor. AB - Interleukin (IL)-3, IL-5 and granulocyte-macrophage colony-stimulating factor (GM CSF) regulate proliferation, differentiation and apoptosis of target cells. Receptors for these cytokines consist of a cytokine-specific alpha subunit and a common shared beta c subunit. Tyrosine phosphorylation of the beta c is thought to play a critical role in mediating signal transduction events. We have examined the effect of mutation of beta c tyrosines on the activation of multiple signal transduction pathways. Activation of protein kinase B (PKB) required JAK2 and was inhibited by dominant-negative phosphatidylinositol 3-kinase (P13K). Overexpression of JAK2 was sufficient to activate both protein kinase B (PKB) and extracellular regulated kinase-1 (ERK1). Tyrosine 577 and 612 were found to be critical for the activation of PKB and ERK1, but not activation of STAT transcription factors. Activation of both PKB and ERK have been implicated in the regulation of proliferation and apoptosis. We generated GM-CSFR stable cell lines expressing receptor mutants to evaluate their effect on these processes. Activation of both PKB and ERK was perturbed, while STAT activation remained unaffected. Tyrosines 577 and 612 were necessary for optimal proliferation, however, mutation of these tyrosine residues did not affect GM-CSF mediated rescue from apoptosis. These data demonstrate that while phosphorylation of beta c tyrosine residues 577 and 612 are important for optimal cell proliferation, rescue from apoptosis can be mediated by alternative signalling routes apparently independent of PKB or ERK activation. PMID- 10362353 TI - Evidence for a p23 caspase-cleaved form of p27[KIP1] involved in G1 growth arrest. AB - p27[KIP1] (p27) is a cyclin dependent kinase inhibitor, involved in the negative regulation of G1 progression in response to a number of anti-proliferative signals. In this study we show, in growing mouse hybridoma (7TD1) and human myeloma (U266) cell lines, that p27 is highly expressed but slightly upregulated when cells are arrested, regardless to the phases of the cell cycle. In contrast, the specific blockade of these cells in early G1 phase reveals the induction of a protein of 23 kDa (p23) specifically recognized by polyclonal anti-p27 antibodies raised against the NH2 terminal part of p27 but not by anti-p21[CIP1] antibodies. Experiments using caspase inhibitors strongly suggest that p23 results from the proteolysis of p27 by a 'caspase-3-like' protease. This cleavage leads to the cytosolic sequestration of p23 but does not alter its binding properties to CDK2 and CDK4 kinases. Indeed, p23 associated in vivo with high molecular weight complexes and coprecipitated with CDK2 and CDK4. We demonstrate by transfection experiments in SaOS-2 cells that p23 induces a G1 phase growth arrest by inhibition of cyclin/CDK2 activity. In summary we describe here a caspase-cleaved form of p27, induced in absence of detectable apoptosis and likely involved in cell cycle regulation. PMID- 10362355 TI - Involvement of the adapter protein CRKL in integrin-mediated adhesion. AB - CRKL, an SH2-SH3-SH3 adapter protein, is one of the major tyrosine phosphoproteins detected in primary leukemic neutrophils from patients with CML. CRKL binds directly to BCR/ABL through its N-terminal SH3 domain, suggesting it may be involved in BCR/ABL signal transduction. However, the biological function of CRKL in either normal or leukemic cells is still largely unknown. In this study, we have examined the effects of overexpressing full length or deletion mutants of CRKL in hematopoietic cell lines. Full length, SH2- and SH3(N)-domain deletion mutants of CRKL were transfected into an interleukin-3-dependent hematopoietic cell line, Ba/F3, and 3-5 individual sublines which stably overexpressed each transgene were obtained [Ba/F-CRKL, Ba/F-CRKL deltaSH2, and Ba/F-CRKL deltaSH3(N)]. The growth properties of these transfected cells in the presence or absence of IL-3 were not different from mock transfected or untransfected Ba/F3 cells. However, Ba/F3 cells overexpressing full length CRKL, but not deletion mutants of CRKL, were found to have an increase in their ability to bind to fibronectin-coated surfaces. Further, expression of full length, but not deltaSH2- or deltaSH3-CRKL deletion mutants, was found to alter cell morphology on fibronectin-coated plates, an effect which was further enhanced by certain kinds of stress stimuli, such as ionizing radiation. Similar results were obtained when CRKL was transiently overexpressed in Ba/F3 cells, and were also obtained in a second IL-3 dependent hematopoietic cell line, 32Dcl3. Adhesion to fibronectin was blocked by anti-beta1 integrin monoclonal antibody, but overexpression of CRKL did not affect surface expression of beta1 integrins, nor did it spontaneously induce expression of the beta1 integrin 'activation' epitope recognized by the 9EG7 monoclonal antibody. These data suggest a role for CRKL in signaling pathways which regulate adhesion to fibronectin. PMID- 10362356 TI - Contribution of Src and Ras pathways in FGF-2 induced endothelial cell differentiation. AB - We have examined fibroblast growth factor (FGF) receptor-1 mediated signal transduction in differentiation of endothelial cells (EC). The activated FGFR-1 couples to Ras through two adaptor proteins, FRS2 and Shc. In FGF-2 treated proliferating EC, FRS2 as well as Shc are tyrosine phosphorylated and interact with Grb2. In contrast, in FGF-2 treated differentiating cells, Shc, but not FRS2, is engaged in Grb2-interactions. Sustained MAP kinase activity has previously been implicated in differentiation. In FGF stimulated proliferating and differentiating endothelial cells, the MAP kinase Erk2 is activated in a sustained manner. Inhibition of MEK and MAP kinase activity by PD98059 treatment of cells, still allows EC tube formation. The FGFR-1 mediates activation of protein kinase C (PKC) through direct binding and activation of phospholipase C gamma (PLC-gamma), and has also been shown to activate the cytoplasmic tyrosine kinase Src. Treatment of the cells with the PKC inhibitor bisindolylmaleimide does not prevent tube formation. In contrast, Src kinase activity is a prerequisite for EC differentiation, since treatment of the cells with PP1, a Src family specific inhibitor, abrogates tube formation. In differentiating EC, FGF-2 induces complex formation between Src and focal adhesion kinase (FAK). These data indicate that the Ras pathway is initiated via Shc or FRS2, dependent on the cellular program. Blocking the function of Src family kinases, attenuates differentiation. PMID- 10362357 TI - cbl-3: a new mammalian cbl family protein. AB - We have cloned a new human gene, cbl-3, which encodes a protein with marked homology to the cbl family of proteins. The predicted protein encoded by this gene retains the conserved phosphotyrosine binding domain (PTB) in the N-terminal and the zinc finger but is significantly shorter (MW 52.5 kDa) than the other mammalian cbl proteins. The protein lacks the extensive proline rich domain and leucine zipper seen in c-cbl and cbl-b and structurally most resembles the C. elegans and Drosophila cbl proteins. The gene is ubiquitously expressed with highest expression in the aerodigestive tract, prostate, adrenal gland, and salivary gland. The protein is phosphorylated and recruited to the EGFR upon EGF stimulation and inhibits EGF stimulated MAP kinase activation. In comparison to the other mammalian cbl proteins (e.g. cbl-b), cbl-3 interacts with a restricted range of proteins containing Src Homology 3 regions. An alternatively spliced form of the cbl-3 protein was also identified which deletes a critical region of the PTB domain and which does not interact with the EGFR nor inhibit EGF stimulated MAP kinase activation. These data demonstrate that cbl-3, a novel mammalian cbl protein, is a regulator of EGFR mediated signal transduction. PMID- 10362359 TI - Repression of an alternative mechanism for lengthening of telomeres in somatic cell hybrids. AB - Some immortalized cell lines maintain their telomeres in the absence of detectable telomerase activity by an alternative (ALT) mechanism. To study how telomere maintenance is controlled in ALT cells, we have fused an ALT cell line GM847 (SV40 immortalized human skin fibroblasts) with normal fibroblasts or with telomerase positive immortal human cell lines and have examined their proliferative potential and telomere dynamics. The telomeres in ALT cells are characteristically very heterogeneous in length, ranging from very short to very long. The ALT x normal hybrids underwent a rapid reduction in telomeric DNA and entered a senescence-like state. Immortal segregants rapidly reverted to the ALT telomere phenotype. Fusion of ALT cells to telomerase-positive immortal cells in the same immortalization complementation group resulted in hybrids that appeared immortal and also exhibited repression of the ALT telomere phenotype. In these hybrids, which were all telomerase-positive, we observed an initial rapid loss of most long telomeres, followed either by gradual loss of the remaining long telomeres at a rate similar to the rate of telomere shortening in normal telomerase-negative cells, or by maintenance of shortened telomeres. These data indicate the existence of a mechanism of rapid telomere deletion in human cells. They also demonstrate that normal cells and at least some telomerase-positive immortal cells contain repressors of the ALT telomere phenotype. PMID- 10362358 TI - Constitutive activation of stimulatory guanine nucleotide binding protein (G(S)alphaQL)-mediated signaling increases invasiveness and tumorigenicity of PC 3M prostate cancer cells. AB - An abnormal stimulation of cAMP signaling cascade has been implicated in various human carcinomas. Since the agents activating G(S)alpha-mediated signaling pathways have been shown to increase in vitro proliferation of prostate cancer cells, present studies examined the G(S)alpha-mediated signaling in tumorigenicity and invasiveness of PC-3M prostate cancer cells. PC-3M cells were stably transfected with plasmids containing either wild type (G(S)alpha-WT) or constitutively active (gsp mutant of G(S)alpha or G(S)alpha-QL) cDNAs. The stable transfectants were then tested for: (1) colony formation in soft agar; (2) cell migration and penetration of basement matrix in an in vitro invasion assay; and (3) the ability to form tumors and metastases in nude mice. PC-3M cells expressing G(S)alpha-QL protein displayed 15-fold increase in their ability to migrate and penetrate the basement membrane as compared to parental PC-3M cells or those expressing G(S)alpha-WT. G(S)alpha-QL transfectants also displayed a dramatically greater rate of growth in soft agar, and greater tumorigenicity and metastasis forming ability when orthotopically implanted in nude mice. All mice receiving PC-3M cells produced primary tumors within 5 weeks after implantation. However, the cells expressing G(S)alpha-QL displayed a significantly faster tumor growth as assessed by prostate weight (greater than 20-fold as compared to PC-3M cells), and produced metastases in kidneys, lymph nodes, blood vessels, bowel mesentery and intestine. Interestingly, expression of G(S)alpha-WT reduced the ability of PC-3M cells to form tumors in nude mice. These results suggest that persistent activation of G(S)alpha-mediated signaling cascade can dramatically accelerate tumorigenesis and metastasizing ability of prostate cancer cells. PMID- 10362360 TI - Etk/Bmx, a PH-domain containing tyrosine kinase, protects prostate cancer cells from apoptosis induced by photodynamic therapy or thapsigargin. AB - Prostate carcinoma (PCA) is the most frequently diagnosed malignancy in American men. PCA at advanced stages can both proliferate abnormally and resist apoptosis. Among the many known signal transduction pathways, phosphatidylinositide-3'OH kinase (PI3-kinase) has been shown to play an important role in cell survival and resistance to apoptosis. In this study, we investigate the involvement of Etk/Bmx, a newly discovered tyrosine kinase that is a substrate of PI3-kinase, in protection of prostate cancer cells from apoptosis. Parental LNCaP cells and two derivative cell lines, one overexpressing wild type Etk (Etkwt) and the other expressing a dominant negative Etk (EtkDN), were used to study the function of Etk. The cells were treated with photodynamic therapy (PDT), a newly approved cancer treatment which employs a photosensitizer and visible light to produce an oxidative stress in cells, often leading to apoptosis. Our results indicate that PDT induces apoptosis in LNCaP cells, as measured by DNA fragmentation and by cleavage of poly(ADP-ribose) polymerase (PARP), and moreover, the extent of apoptosis was much reduced in Etkwt cells as compared to LNCaP or EtkDN cells. Assay of overall cell viability confirmed that Etkwt cells were considerably less sensitive to PDT than were the parental LNCaP or EtkDN cells. Similar results were found in response to thapsigargin (TG). A specific inhibitor of PI3-kinase, LY294002, abolished Etk activity and markedly increased TG-induced PARP cleavage. The results suggest that Etk/Bmx is an efficient effector of PI3-kinase and that the newly described PI3-kinase/Etk pathway is involved in the protection of prostate carcinoma cells from apoptosis in response to PDT or TG. PMID- 10362361 TI - Differential signaling by alternative HGF isoforms through c-Met: activation of both MAP kinase and PI 3-kinase pathways is insufficient for mitogenesis. AB - HGF/NK2, a naturally occurring truncated HGF isoform, antagonizes the mitogenic and morphogenic activities of full length HGF, but stimulates cell scatter, or the motogenic response to HGF. We studied postreceptor signaling by these HGF isoforms in the human breast epithelial cell line 184B5, and in murine myeloid progenitor 32D cells transfected with c-Met, the human HGF receptor (32D/c-Met). HGF stimulated DNA synthesis in 184B5 and 32D/c-Met cells, while HGF/NK2 was mitogenically inactive, despite the ability of HGF/NK2 to stimulate c-Met autophosphorylation, mitogen-activated protein kinase (MAPK), and phosphatidylinositol 3-kinase (PI3K) in both cell systems. In 184B5 cells, HGF stimulated sustained MAPK activation, while activation by HGF/NK2 declined rapidly. In contrast, both isoforms activated MAPK with rapidly attenuated kinetics in 32D/c-Met cells. In both cell systems the increased motility observed in response to either HGF or HGF/NK2 treatment was more potently blocked by the PI3 kinase inhibitor wortmannin, than by PD98059, an inhibitor of MAPK kinase (MEK1). These data suggest that (1) alternative HGF isoforms signaling through c Met generate both common and distinct biological responses, (2) the extent and duration of ligand-stimulated c-Met and MAPK activities are dependent on the cellular context and are not predictive of mitogenic signaling, and (3) in at least some HGF target cells, the activation of both MAPK and PI3K signaling pathways is insufficient for mitogenesis elicited through c-Met. PMID- 10362362 TI - Telomerase activity in ductal carcinoma in situ and invasive breast cancer. AB - The increasing number of breast carcinoma in situ detected by screening procedures makes it imperative to develop improved markers to stratify the risk of invasive cancer. Telomerase is detectable in invasive cancer, but not in normal tissues. We have microdissected frozen tissue blocks containing both DCIS and invasive cancer to assay the telomerase activity of these two lesions. The 46 available cases of concurrent DCIS and invasive breast cancer resulted in 43 DCIS samples and 38 invasive cancer samples adequate for analysis. Seventy per cent of the DCIS and all invasive cancer samples tested had detectable telomerase activity. In addition, we analysed telomerase activity in ten cases of DCIS that were not associated with invasive cancer, and detected telomerase activity in seven (70%). Mixing experiments showed no evidence of telomerase inhibitors in telomerase negative samples. Furthermore, periductal inflammatory infiltrates were shown to be a potential confounding source of telomerase activity. Since DCIS lesions appear to be heterogeneous with respect to telomerase activity, and telomerase activation appears to precede the development of invasive cancer, telomerase activity may be a useful adjunct in stratifying the risk of developing invasive breast cancer in patients with DCIS. PMID- 10362363 TI - Mutational analysis of p73 and p53 in human cancer cell lines. AB - p73 is a candidate tumor suppressor gene with substantial DNA and protein homology to the p53 tumor suppressor gene. We have investigated two hypotheses: (a) p73 is mutated in diverse types of human cancer, and (b) p73 is functionally redundant with p53 in carcinogenesis so that mutations would be exclusive in these two genes. The entire coding region and intronic splice junctions of p73 were examined in 54 cancer cell lines. Three lung cancer cell lines contained mutations that affected the amino acid sequence. One amino acid substitution was in a region with homology to the specific DNA binding region of p53 and two microdeletions were outside the region of homology. Two of the cell lines with p73 mutations also carried p53 mutations. Although our results are inconsistent with the two hypotheses tested, p73 mutations may contribute infrequently to the molecular pathogenesis of human lung cancer. PMID- 10362364 TI - Mutations of a novel human RAD54 homologue, RAD54B, in primary cancer. AB - Association of breast tumor susceptibility gene products BRCA1 and BRCA2 with the RAD51 recombination protein suggested that cancer could arise through defects in recombination. The identification of NBS1, responsible for Nijmegen breakage syndrome, from the MRE11/RAD50 recombination protein complex also supports this hypothesis. However, our mutation analysis revealed that known members of the RAD52 epistasis group are rarely mutated in human primary cancer. Here we describe the isolation of a novel member of the SNF2 superfamily, characterized with sequence motifs similar to those in DNA and RNA helicases. The gene, designated RAD54B, is significantly homologous to the RAD54 recombination gene. The expression of RAD54B was high in testis and spleen, which are active in meiotic and mitotic recombination. These findings suggest that RAD54B may play an active role in recombination processes in concert with other members of the RAD52 epistasis group. RAD54B maps to human chromosome 8q21.3-q22 in a region associated with cancer-related chromosomal abnormalities. Homozygous mutations at highly conserved positions of RAD54B were observed in human primary lymphoma and colon cancer. These findings suggest that some cancers arise through alterations of the RAD54B function. PMID- 10362365 TI - Mutations in the RAD54 recombination gene in primary cancers. AB - Association of a recombinational repair protein RAD51 with tumor suppressors BRCA1 and BRCA2 suggests that defects in homologous recombination are responsible for tumor formation. Also recent findings that a protein associated with the MRE11/RAD50 repair complex is mutated in Nijmegen breakage syndrome characterized by increased cancer incidence and ionizing radiation sensitivity strongly support this idea. However, the direct roles of BRCA proteins and the protein responsible for NBS in recombinational repair are not clear though they are associated with the recombinational repair complexes. Since RAD51 forms a complex with other members of the RAD52 epistasis group and with BRCA proteins, it is reasonable to ask if alterations of members of the RAD52 epistasis group lead to tumor development. Here we describe missense mutations at functional regions of RAD54 and the absence of the wild-type RAD54 expression resulting from aberrant splicing in primary cancers. Since RAD54 is a recombinational protein associated with RAD51, this is the first genetic evidence that cancer arises from a defect in repair processes involving homologous recombination. PMID- 10362366 TI - Plasma vasoactive peptides after acute myocardial infarction in relation to left ventricular dysfunction. AB - We measured plasma concentrations of vasoactive peptides in 32 patients with acute myocardial infarction and evaluated their value as markers of left ventricular dysfunction. Plasma levels of atrial natriuretic peptide (ANP), the N terminal fragment of proANP (NT-proANP), B-type natriuretic peptide (BNP) and endothelin-1 were measured serially by radioimmunoassays. The infarct size was estimated from the creatine kinase MB release curve. Coronary angiography and left ventricular cineangiography were performed in all patients during hospitalization and 6 months later in 15 patients. Myocardial infarction caused an increase in vasoactive peptides, the highest values for ANP (36.5+/-6.79 pmol/l), NT-proANP (1130+/-170 pmol/l) and endothelin-1 (9.72+/-0.68 pmol/l) being found on admission and those for BNP (56.0+/-7.13 pmol/l) on Day 2. Plasma levels of natriuretic peptides were dependent on infarct size, its location and degree of myocardial dysfunction and that of BNP also on infarct artery patency. Plasma endothelin-1 level was higher in patients with TIMI 3 than TIMI 0-2 flow. Plasma vasoactive peptides remained elevated during the 6-month follow-up period and they were dependent on the degree of myocardial dysfunction. BNP measured on any day of hospitalization showed the best correlation with ejection fraction measured during the acute phase of infarction or at 6 months. The results show that BNP is the best indicator of left ventricular dysfunction after myocardial infarction and its reliability is not dependent on the time point of measurement. PMID- 10362367 TI - Great expectations from a different approach to the treatment of acute myocardial infarction : cytoprotection. PMID- 10362368 TI - Coronary artery flow ten weeks after myocardial infarction or unstable angina: effects of combined warfarin and aspirin therapy. AB - Forty-three patients presenting with unstable angina or myocardial infarction were randomised double blind to warfarin [target international normalised ratio (INR), 2.0 to 2.5] and aspirin (150 mg) daily or placebo plus aspirin (150 mg) daily. Coronary flow was assessed with the thrombolysis in myocardial infarction (TIMI) flow grade and corrected TIMI frame count (CTFC). Coronary artery flow was reduced (higher CTFC) at baseline in culprit arteries (mean +/-SD, 37.1+/-15.4 frames) compared to nonculprit arteries (22.5+/-6.7 frames, P<0.0001). In patients with a patent artery at follow-up, coronary flow was unchanged after ten weeks of warfarin and aspirin (-2.0+/-19.9 frames) or aspirin alone (3.8+/-10.4 frames, P = 0.20). Patients randomised to aspirin alone were more likely to progress to total occlusion [aspirin, 7 of 19 (37%) vs. warfarin and aspirin, 1 of 24 (4%); P = 0.01). Higher baseline culprit artery CTFC was also associated with an increased risk of late occlusion [+10 frames; odds ratio (OR), 1.65; 95% CI, 1.01 to 2.33]. Coronary flow remained impaired ten weeks after presentation with myocardial infarction or unstable angina. Combination warfarin and aspirin therapy did not improve flow in vessels that remained patent but did reduce the risk of progression to occlusion. PMID- 10362369 TI - Balloon dilatation for critical pulmonary stenosis. AB - This study was conducted to investigate the outcome of balloon valvuloplasty for critical pulmonary stenosis in young infants. During a 6.2-year period between January 1992 and February 1998, 34 infants with critical pulmonary stenosis, aged 1 to 58 days (16.8+/-16.6 days), underwent attempted balloon valvuloplasty in this institution. The procedure was accomplished in 28 patients, but failed in six. Surgical pulmonary valvotomy was performed in the six patients with one mortality. Immediately following valvuloplasty, the mean right ventricular systolic pressure decreased from 109.2+/-28.6 to 55.1+/-23.6 mm Hg in the 28 patients (P<0.01). The mean pressure gradient decreased from 85.6+/-29.4 to 26+/ 21.4 mm Hg (P<0.01). However, one who had a severely hypoplastic right ventricle requiring prolonged prostaglandin E1 infusion after valvuloplasty underwent a right ventricular outflow tract patch. After a follow-up period ranging from 2 months to 6.4 years (30.5+/-19.1 months), one patient developed recurrent pulmonary stenosis and underwent a repeated balloon valvuloplasty. Of the 27 patients (79%) with a definitive success of balloon valvuloplasty, the mean pressure gradient estimated with Doppler echocardiography at most recent follow up was 15.2+/-6.8 mm Hg. Therefore, balloon valvuloplasty is the procedure-of choice for critical pulmonary stenosis. Surgery should be reserved for those with unsuccessful balloon valvuloplasty. PMID- 10362370 TI - Age, body mass index and 2-hour plasma glucose are the major determinants of blood pressure in Chinese women newly diagnosed to have glucose intolerance. AB - Hypertension and diabetes frequently coexist and greatly increase cardiovascular risk. There are relatively few data on the relationship between blood pressure and plasma glucose in newly diagnosed diabetic subjects especially in Chinese. We examined the glycaemic status, blood pressure profiles and other clinical and biochemical characteristics in 1298 Hong Kong Chinese women. These women were referred to the Diabetes and Endocrine Center of the Prince of Wales Hospital for screening of diabetes. The reasons for referral included a positive family history of diabetes or a history of gestational diabetes. Of the 1298 subjects, 836 (64.4%) had normal oral glucose tolerance test, 284 (21.9%) had impaired glucose tolerance and 178 (13.7%) had diabetes. Compared to non-diabetic subjects, the odds ratio (95% confidence interval) of having hypertension in subjects with impaired glucose tolerance or diabetes were 2.83 (1.90, 4.23) (P<0.001) and 5.94 (3.94, 8.96) (P<0.001), respectively. When analyzed as a continuous variable by age-adjusted partial correlation coefficients, systolic blood pressure was correlated with body mass index, fasting and 2-h plasma glucose, while diastolic blood pressure was correlated with body mass index. Using age, body mass index, fasting and 2-h plasma glucose, glycated hemoglobin, cholesterol, triglyceride and smoking as independent variables in multivariate analysis, hypertension was independently related to age, body mass index and 2-h plasma glucose. In conclusion, increased blood pressure was common in Hong Kong Chinese women who were newly diagnosed to have glucose intolerance. Apart from age and body mass index, plasma glucose was an independent determinant for blood pressure in these subjects. PMID- 10362371 TI - Quantitation of Doppler color flow jet areas for mitral regurgitation in children: angiographic correlation. AB - Eighteen patients with chronic isolated rheumatic mitral regurgitation aged between 7 and 19 years (mean age +/-SD, 12.69+/-3.47 years) were analyzed with color Doppler imaging. Sixteen patients were performed cardiac catheterization within 24 h. Jets were classified as eccentric and central. Regurgitant jet area and its ratio to left atrial area and body surface area were measured by Doppler color flow imaging. Regurgitant volume and regurgitant fractions were calculated with angiography. There was a good correlation between regurgitant jet area and angiographic grade of mitral regurgitation (P<0.01). The correlation between regurgitant jet area/left atrial area ratios and angiographic grade of mitral regurgitation was limited (P<0.01). There was excellent correlation between regurgitant jet area/body surface area and angiographic regurgitant fraction (r = 0.85; P<0.001). There was also a good correlation between regurgitant jet area and regurgitant fraction (r = 0.82; P<0.001). However, the relation of regurgitant jet area/left atrial area to regurgitant fraction was weak (r = 0.72; P<0.01). In conclusion, the measurement of regurgitant fraction and its ratios to left atrial area and body surface area by color Doppler flow imaging can predict the angiographic severity in children who have even eccentric regurgitant jets. PMID- 10362372 TI - Atrial fibrillation after coronary artery bypass surgery: P wave signal averaged ECG, clinical and angiographic variables in risk assessment. AB - BACKGROUND: Atrial fibrillation (AF) is a commonly encountered arrhythmia and occurs in up to 40% of patients after coronary artery bypass surgery (CABG). The preoperative signal averaged ECG (SAECG) P wave may be useful indicator of AF after CABG. We prospectively analyzed the predictive value of SAECG P wave compared to clinical variables. METHODS: Fifty-three patients with coronary artery disease undergoing first elective CABG were enrolled. All patients had P wave specific SAECG, standard 12 lead ECG, ejection fraction and left atrial posteroanterior diameter from the echocardiogram within the 24 h before surgery. From the SAECG P wave, filtered P wave duration was measured. Lead II P wave duration, left atrial enlargement and left ventricular hypertrophy were determined from standard ECG. Patients were continuously monitored during their postoperative period and serial ECGs were taken. RESULTS: During an observation period of up to 16 days, 19 (35.8%) patients developed AF 2.8+/-1.3 days after CABG. Patients with AF more often had left atrial enlargement (LAE) on ECG (P = 0.041) and right coronary artery (RCA) lesion (P = 0.0034). The filtered P wave duration on the SAECG was significantly longer in the AF patients than those without AF (129.7+/-13.2 ms versus 113.9+/-9.0 ms, P = 0.001). Logistic regression analysis identified independent predictors, estimated adjusted relative risk (95% confidence interval) of AF: with LAE, the relative risk was 2.72 (1.13-5.82), RCA lesion, the relative risk was 3.06 (1.45-6.45) and SAECG P wave duration >122.3 ms, the relative risk was 4.58 (2.11-9.97). The occurrence of AF was predicted by electrocardiographically determined left atrial enlargement with a sensitivity of 36%, specificity of 88%, positive predictive accuracy of 63%, negative predictive accuracy of 71%. If presence of right coronary artery lesion was evaluated these values were 63%, 79%, 63%, 79% subsequently. P wave duration >122.3 ms had a sensitivity of 68%, specificity of 88%, positive predictive accuracy of 76%, negative predictive accuracy of 83%. If both P wave >122.3 ms and presence of right coronary artery lesion were combined, these values were 47%, 94%, 81%, 76% subsequently. CONCLUSION: The predictors of AF after CABG were left atrial enlargement on standard 12 lead ECG, RCA lesion and SAECG P wave duration. Among these predictors, SAECG P wave duration was the best predictor of AF after CABG. PMID- 10362373 TI - Transarterial occlusion of patent ductus arteriosus with Gianturco coils in pediatric patients: a preliminary result in central Taiwan. AB - OBJECTIVE: We wish to present the preliminary result of transarterial occlusion of patent ductus arteriosus (PDA) with Gianturco coils in pediatric patients in central Taiwan. MATERIALS AND METHODS: We attempted occlusion of PDA with Gianturco coils in a total of 26 consecutive patients, 13 infants and 13 children, 23 female and three male, between July 1 1997 to September 30 1998. Median patient age was 2.57 years (from 0.25 to 14.02 years old). Median patient weight was 10.8 kg (4.0 to 36.0 kg). Premature babies with PDA, full-term babies who were less than three months old and patients who had other congenital heart disease were not included in this study. All PDAs were approached transarterially from the femoral artery. Coils were selected to provide a helical diameter that was twice or more the minimum ductus diameter and a length approximating five loops. In five patients who had a PDA diameter > or =3.5 mm, we used a snare technique to assist coil delivery beforehand, and to test coil stability, or to retrieve coil that had migrated to the pulmonary artery afterwards. Physical auscultation, chest radiographs and echocardiography with color Doppler were done in all patients within 24 h, and one, two, three, six and 12 months after coil occlusion. RESULTS: The median ductus minimum diameter was 2.3 mm (range, 1.0 to 4.7 mm). Fifteen patients had the megaphone type (type A), four had the window type (type B), five had the tubular type (type C), one had the aneurysmal type (type D) and one had the elongated conical type (type E). Twenty-one patients underwent single coil occlusion and five had multiple coils occlusion. Twenty-one patients had immediate angiographic closure of the ductus and disappearance of heart murmur at 15 min after the procedure. Dark-brown urine (hemoglobinuria) was found in one patient, 10 h after the first procedure, due to a mild residual ductal shunt. Two more coils were implanted in a second procedure that was performed within 24 h, and the ductus was completely occluded. The dark-brown urine regressed. At one month follow-up, four patients had mild residual ductal shunts, which were completely occluded by one more coil in three patients and by two more coils in the other patient. Malpositioned coils were deployed in five patients immediately after the procedure. In total, the closure rate at 15 min, within 24 h, and at one, two, three, six and 12 months were 81, 85, 85, 100, 100, 100 and 100%, respectively. In one year of follow-up, there was no instance of coil migration, ductus reopening or stenosis of the left pulmonary artery. CONCLUSIONS: Transarterial occlusion of PDA, with a Gianturco coil having approximately five loops, can be effectively achieved in patients with a minimum ductus diameter up to 4.7 mm. In patients with a ductus of more than 3.5 mm, the snare-assisted technique was employed advantageously to control coil delivery with accuracy and stability. Coil malposition or migration can be easily retrieved using a 10-mm Nitnol snare catheter. Hemoglobinuria, due to intravascular hemolysis, may regress within 24 h after the second attempt at coil implantation. PMID- 10362374 TI - Coronary artery aneurysm formation after balloon angioplasty and stent implantation. AB - This study describes coronary angiographic and intravascular ultrasound evaluation of late coronary artery aneurysms after percutaneous balloon angioplasty and bailout stent implantation. Intravascular ultrasound distinguishes true aneurysms from pseudoaneurysms. The discussion is focused on the etiology and prognosis of this rare complication. PMID- 10362375 TI - Do we have a less severe form of Kawasaki disease or is it the gammaglobulin effect? AB - This study was undertaken to evaluate the incidence of coronary artery aneurysms (CAA) in Kawasaki disease (KD). We reviewed the clinical and echocardiographic findings of 135 children who presented to our center with KD between December 1986 and December 1997. The age of onset ranged between 3months to 13 years (median 2 years). The male to female ratio was 1.54:1. All patients received intravenous Gammaglobulin (IVGG) during the acute stage. The echocardiogram, which was done between 2-3 weeks of the onset of fever, was normal in 106 patients (78.5%). Follow-up studies over a period of 6 months to 1 year remained normal. Minimal right or left coronary artery wall ectasia without dilatation or aneurysm formation was seen in 16 (11.85%). Follow-up of these patients showed disappearance of these changes over 6 weeks to 6 months. One patient (0.74%) had generalised dilatation of all the coronary arteries during the acute stage. This has normalized over a period of 9 months. A total of 10 (7.4%) had CAA during the acute stage. On follow-up of 8 of these patients for an average 3 months to 1.5 years all CAA regressed completely. One patient had residual Giant CAA after 1 year follow-up. One patient with CAA was lost to follow-up. One patient (0.74%) had pericardial effusion and another one (0.74%) had mitral incompetence during the acute stage only, both had no coronary involvement. None of our patients had cardiac failure, arrhythmia, myocardial infarction or death. We conclude that coronary artery changes due to KD are less common and less severe in our patients than those seen in other studies. We speculate that this can be related partly to the early administration of IVGG. The difference in incidence of CAA secondary to KD among different racial groups warrants more detailed genetic studies. PMID- 10362376 TI - The changes of circulating tumor necrosis factor levels in patients with congestive heart failure influenced by therapy. AB - Recent studies suggest that tumor necrosis factor-alpha (TNF-alpha) plays an important role in the pathogenesis of congestive heart failure and that drugs used in the treatment of heart failure have modulation effects on the production of TNF-alpha. To examine an alteration of circulating TNF-alpha concentration in patients with severe chronic heart failure after improving heart function and investigate the influence of agents on circulating TNF-alpha concentrations, we measured the plasma levels of TNF-alpha by enzyme linked immunoabsorbent assay in 31 patients and evaluated their heart functions before and after 72 h of therapy. The results showed that circulating TNF-alpha concentrations significantly decreased after therapy (from 124.36+/-14.85 pg/ml to 93.84+/-13.57 pg/ml, P<0.001). The circulating TNF-alpha concentrations of patients (n = 22) whose heart function was improved one class or more after therapy declined significantly (from 127.51+/-20.78 pg/ml to 91.54+/-18.56 pg/ml, P<0.01) but this situation did not exist in patients (n = 9) whose heart functions had no or little improvement. All patients were divided into three groups according to their management: 'group A' (n = 14) who received milrinone and angiotensin converting enzyme inhibitors (ACEI), 'group B' (n = 6) who received milrinone but not ACEI and 'group C' (n = 11) who received ACEI and dobutamine but not milrinone. The circulating TNF-alpha concentration of patients in group A significantly declined (from 126.68+/-26.04 pg/ml to 95.92+/-24.79 pg/ml, P<0.01). No statistical significance of change of TNF-alpha concentration was found in patients in group B or group C, although a tendency of decline existed (from 119.92+/-34.72 pg/ml to 84.33+/-30.70 pg/ml and from 123.83+/-19.50 pg/ml to 96.37+/-16.62 pg/ml, respectively). These findings support that decreased plasma TNF-alpha level accompanies the improvement of heart function. This phenomenon may be explained by the special abilities of agents, such as ACEI and milrinone, to inhibit the TNF-alpha production. PMID- 10362377 TI - Acute and chronic phase platelet aggregability studies in Chinese patients after implantation of a permanent transvenous pacemaker. AB - It has been suggested that the incidence of thromboembolic events always increases in patients after insertion of a transvenous pacemaker. Blood samples from twenty consecutive patients (fifteen males and five females) before and after pacemaker implantation was retained for platelet aggregability studies which were analyzed separately with ADP, collagen, epinephrine and arachidonic acid. The maximal amplitude of platelet aggregatory curve was detected by an aggregometer. The samples collected the day before pacemaker implantation (day 0) were used as self-control. Day 1 and day 3 after pacemaker implantation were defined as the acute phase, while day 30 was defined as the chronic phase. The maximal amplitude of platelet aggregatory curve was observed to be lowest on day 1 and then return to normal on day 3 and day 30. The results of platelet aggregability, however, showed no significant difference (P>0.05) between self control and post-implantation samples. In conclusion, there was no significant change in platelet aggregability for either acute or chronic phases after pacemaker implantation. Antiplatelet medications may not be necessary for the prevention of thromboembolic events after the implantation of a pacemaker. PMID- 10362378 TI - Acquired thromboatheromatous coarctation of the aorta: acquired coarctation of the aorta. AB - BACKGROUND: Atheromatosis of the thoracic aorta and aortic arch is a well established source of systemic embolism. Acquired atheromatous coarctation of the aortic arch is a rare finding and not well documentated so far. CASE REPORT AND FINDINGS: Two patients presenting with intermittent claudication of the lower extremities were identified as having thromboatheromatous coarctation of the aortic arch as visualized by magnetic resonance tomography, fast CT scan, transesophageal echocardiography, cardiac catheterization and aortography. All findings including invasive hemodynamics resembled congenital coarctation of the aorta. One patient was treated surgically, while the other refused surgery and received long-term anticoagulation. CONCLUSION: Atheromatosis of the thoracic aorta and aortic arch not only cause systemic embolism, but may lead to the clinical and hemodynamic picture of coarctation of the aortic arch. PMID- 10362379 TI - Combined dynamic cardiomyoplasty and transvenous implantable cardioverter defibrillator system. AB - A transvenous implantable cardioverter defibrillator (ICD) was implanted in a patient with drug refractory ventricular fibrillation who had undergone latissimus dorsi cardiomyoplasty. The skeletal muscle was stimulated by a pulse train generator (cardiostimulator) implanted at the time of cardiomyoplasty. With proper programming of the devices neither adverse ICD-cardiostimulator interactions nor device malfunction were observed. Thus, the combined implantation of a cardiostimulator and an ICD is a feasible and safe therapeutic option. PMID- 10362380 TI - Endocarditis due to Acinetobacter lwoffi on native mitral valve. AB - Endocarditis due to Acinetobacter is a rare pathology with high mortality, reported mainly in hospitalized patients with predisposing risk factors. This is the second case of endocarditis due to Acinetobacter reported in our country in the last 10 years. PMID- 10362381 TI - Atrial septal aneurysm. Is it a benign finding? AB - We report our clinical experience in a single centre with 7 cases of atrial septal aneurysm (ASA) diagnosed by transthoracic echocardiography (TTE) between 1989-1996. They did not present any clinical event compatible with cardiogenic embolism after a five years mean follow-up period. ASA is recognized as a potential source of cardiogenic embolism [2] based on some retrospective and selection biased studies. PMID- 10362382 TI - Double-lumen aortic arch with anomalous left pulmonary artery origin from the main pulmonary artery--bilateral persistent fifth aortic arch--a case report. AB - Both double-lumen aortic arch (i.e., persistent fifth arch) and anomalous origin of the left pulmonary artery from the ascending aorta (failure of the development of the sixth arch) are rare diseases. They are frequently associated with cardiovascular anomalies. However, the co-occurrence of these two diseases has not been previously reported. We report such a case in a female baby with facial anomalies similar to conotruncal anomaly face syndrome. PMID- 10362383 TI - Treatment of chronic heart failure with perindopril in ethnic Sri Lankan patients. PMID- 10362384 TI - Acute and residual effects of vibratory stimulation on explosive strength in elite and amateur athletes. AB - Fourteen elite and 14 amateur athletes were subjected to vibratory stimulation during bilateral biceps curl exercises of explosive strength exertion. The athletes performed two separate series of three sets of exercises in random order. The second set of one series was administered with superimposed vibration of 44 Hz and an acceleration of about 30 m x s(-2) transmitted through the two arms handle to the arm muscles. The mechanical power of each repetition was measured by the 'Power Teach' instrument. The maximal and mean power values for each set were automatically recorded and shown on the screen. The acute effect was evaluated as the difference between the mean and peak power output in the second (with vibratory stimulation) and first (without vibratory stimulation) sets. Similarly, the residual effect was taken to be the difference between the power values of the third (after vibratory stimulation) and the first (before vibratory stimulation) sets. The results were subjected to a repeated-measures analysis of variance with group as a between-participants factor. The results showed that exercise mode (with vs without vibratory stimulation) resulted in a significant immediate effect for mean power and for maximal power. The factor group (elite vs amateurs) resulted in a significant effect for maximal power only. The increase in explosive strength exertion attributed to vibratory stimulation was 30.1 and 29.8 W (10.4% and 10.2%) for maximal and mean power respectively in the elite group, and 20.0 and 25.9 W (7.9% and 10.7%) respectively in the amateur athletes. Vibratory stimulation resulted in an insignificant residual effect. PMID- 10362385 TI - Perceived discomfort in running: scale development and theoretical considerations. AB - The aim of this study was to develop a discomfort questionnaire to elicit the feelings and thoughts of people engaged in running activities. Ten runners who completed a particularly demanding 9-km run were asked to express their feelings and thoughts during the run they had just completed. These responses were recorded and later used as the first pool of items (k = 36). The questionnaire was then given to 171 runners in different distance races throughout the 1995 competitive season. These responses were analysed using exploratory factor analytic techniques and Rasch probabilistic analysis, as well as traditional reliability and validity procedures. The final version of the questionnaire consisted of 32 items divided into eight correlated subscales: proprioceptive symptoms, leg symptoms, respiratory difficulties, disorientation, dryness and heat, task completion thoughts, mental toughness, and head or stomach symptoms. These eight categories can be collapsed into three global categories suggested by researchers of pain: sensory-discriminative, motivational-affective and cognitive evaluative. Rasch analysis suggested that the motivational-affective and cognitive-evaluative dimensions (i.e. the psychological) are the most experienced (i.e. rated highest). The eight subscales have ecological validity and were found to alter with the demands of different running distances. PMID- 10362386 TI - The effect of a prophylactic dose of flurbiprofen on muscle soreness and sprinting performance in trained subjects. AB - The aim of this study was to examine the effects of a prophylactic dose of a local, transcutaneously administered, non-steroidal anti-inflammatory drug on muscle soreness, muscle damage and sprinting performance in young trained males. Twenty-five subjects aged 19+/-3 years, actively participating in rugby union and field hockey, were familiarized with the test procedure and then divided at random into an experimental group (n = 13) and a control group (n = 12). The experimental group received two patches, each containing 40 mg flurbiprofen (TransAct LAT), 12 h before an exercise bout designed to produce delayed-onset soreness (DOMS). The control group received identical non-medicated placebo patches at the same time. Delayed-onset muscle soreness was induced by an exercise protocol consisting of drop jumps (seven sets of 10 repetitions). Serum creatine kinase activity, muscle soreness, muscle girth and acceleration in a maximal sprint over 30 m were measured before the induction of DOMS and at 12, 24, 48 and 72 h thereafter. Plasma lactate concentration was measured 3 min after the 30-m sprint tests. Subjects in both groups had significantly more pain at 24 and 48 h compared with at 12 and 72 h (P < 0.05; Friedman two-way analysis of variance). Thigh girth and serum creatine kinase did not change throughout the experiment. Although plasma lactate concentrations were elevated after the 30-m sprint, there were no differences between groups or as a result of DOMS. The greatest acceleration occurred between 5 and 10 m. This was not affected by the anti-inflammatory drug or DOMS. In conclusion, the aetiology of the DOMS induced in the trained subjects in this study seems to be independent of inflammatory processes or, more specifically, of increases in prostaglandin synthesis in the muscles. PMID- 10362387 TI - Perceptions of the sport psychologist by female university athletes. AB - In this study we explored the existence of a favourable attitude towards sport psychologists by female athletes in relation to other sport-oriented and mental health professionals. Ninety female student athletes made judgements of similarity between 11 practitioner terms using the triad method. A rank-order task was also completed, where the 11 professionals were ranked on three expertise variables in sporting, mental and physical issues. The results were analysed using (1) the metric scaling procedure of correspondence analysis, (2) cultural consensus analysis and (3) PROperty FITting analysis. A two-dimensional solution provided the best interpretation of the similarity judgements. The correspondence analysis configuration positioned the sport psychologist centrally between a sport-oriented pair and the cluster of mental health professionals. Participants reported adequate consensus on all three expertise variables, which is consistent with the assumptions of Cultural Consensus Theory. Consistent with earlier research, the three variables were salient in the participants' similarity judgements of sport and mental health professionals. Our results suggest the existence of a more favourable perception of the sport psychologist and a distancing from a direct association with mental health practitioners. However, the centrality of the term may indicate a more cloudy distinction as to where the sport psychologist exists in relation to other professionals. PMID- 10362388 TI - Achievement goals, beliefs about the causes of success and reported emotion in post-16 physical education. AB - The main aim of this study was to examine whether goal orientations of male and female adolescents involved in an optional post-16 physical education (PE) programme were related in a conceptually consistent manner with their beliefs about the causes of success in PE. We also determined relationships between these achievement goal-belief dimensions and reported enjoyment and boredom within PE classes. Participants (n = 171) in a sixth-form college PE programme completed an inventory assessing their task and ego goal orientations, beliefs about the determinants of success in PE, and emotion in PE activities at college. Separate factor analyses of goal orientations and beliefs for male and female students revealed two goal-belief dimensions. The first dimension showed ego orientation was linked to the view that ability and deceptive tactics lead to success. The second dimension suggested task orientation was associated with the belief that success is the result of hard work and effort. This task goal-belief factor was found to be more strongly correlated with enjoyment in PE among female students than among males. For boys, the task goal-belief factor was correlated significantly and negatively with boredom in PE, but this was not the case for girls. No significant relationships emerged between the ego goal-belief factor and reported emotion in PE among the male and female participants. Facilitating task involvement and beliefs about causes of success that are fundamentally under personal control may, therefore, promote positive affective experiences in sixth form PE, especially among female students. PMID- 10362389 TI - Horizontal-to-vertical velocity conversion in the triple jump. AB - The aim of this study was to determine the effects of selected factors on horizontal-to-vertical velocity conversion in the triple jump. An understanding of this conversion is important not only for studies on the techniques of the triple jump, but also for other jumping events. Ten elite jumpers were studied. Three-dimensional kinematic data were collected for at least four complete trials in the same competition for each athlete. The loss in horizontal velocity and the gain in vertical velocity during each support phase were calculated for each trial. The loss in horizontal velocity was found to be a linear function of the gain in vertical velocity. The slope of this linear function, A1, is referred to as the horizontal-to-vertical velocity conversion coefficient. The loss in horizontal velocity increased as the gain in vertical velocity increased. The sensitivity of the loss in horizontal velocity to the gain in vertical velocity increased as the magnitude of A1 increased. Further studies are required on the optimum techniques of the triple jump. PMID- 10362390 TI - Effects of cold water immersion on the symptoms of exercise-induced muscle damage. AB - Cryotherapy is an effective treatment for acute sports injury to soft tissue, although the effect of cryotherapy on exercise-induced muscle damage is unclear. The aim of this study was to assess the effects of cold water immersion on the symptoms of exercise-induced muscle damage following strenuous eccentric exercise. After performing a bout of damage-inducing eccentric exercise (eight sets of five maximal reciprocal contractions at 0.58 rad x s(-1)) of the elbow flexors on an isokinetic dynamometer, 15 females aged 22.0+/-2.0 years (mean +/- s) were allocated to a control group (no treatment, n = 7) or a cryotherapy group (n = 8). Subjects in the cryotherapy group immersed their exercised arm in cold water (15 degrees C) for 15 min immediately after eccentric exercise and then every 12 h for 15 min for a total of seven sessions. Muscle tenderness, plasma creatine kinase activity, relaxed elbow angle, isometric strength and swelling (upper arm circumference) were measured immediately before and for 3 days after eccentric exercise. Analysis of variance revealed significant (P < 0.05) main effects for time for all variables, with increases in muscle tenderness, creatine kinase activity and upper arm circumference, and decreases in isometric strength and relaxed elbow angle. There were significant interactions (P<0.05) of group x time for relaxed elbow angle and creatine kinase activity. Relaxed elbow angle was greater and creatine kinase activity lower for the cryotherapy group than the controls on days 2 and 3 following the eccentric exercise. We conclude that although cold water immersion may reduce muscle stiffness and the amount of post exercise damage after strenuous eccentric activity, there appears to be no effect on the perception of tenderness and strength loss, which is characteristic after this form of activity. PMID- 10362391 TI - Effect of endurance training on blood lactate clearance after maximal exercise. AB - The aim of this study was to measure serial changes in the rate of blood lactate clearance (gamma2) in response to sequential periods of training and detraining in four male triathletes aged 22-44 years. There were two major phases of training and taper, each lasting 4-5 weeks (training 1 = 5 weeks, taper 1 = 2 weeks, training 2 = 4 weeks and taper 2 = 2 weeks), in preparation for a triathlon competition. The training stimulus absorbed by each subject was carefully quantified from the duration and intensity of the training exercise. A serial weekly measure of each trainee's physical response to training was evaluated as the peak power, termed a 'criterion performance', developed by a subject during a 30 W x min(-1) ramp cycle ergometer test to exhaustion each week. During 30 min of recovery after this test, 13 samples of venous blood were drawn sequentially from a subject to measure the blood lactate recovery curve. The rate constant of blood lactate clearance was estimated by a non-linear least squares regression technique. In addition, the concurrent time to peak lactate concentration and the peak lactate concentration were also estimated to help define changing lactate kinetics. The criterion performance generally declined throughout each period of incremental training and improved during each taper period, rising iteratively in this way to be clearly above baseline by the end of the second taper. The blood lactate clearance rate increased transiently in early training before declining from the middle of the first training period to the middle of the first taper; thereafter, gamma2 increased above baseline in each trainee throughout the remaining first taper and the major portion of the second training period, decreasing only in the final criterion performance test. The time to peak lactate declined from baseline throughout all phases of training and taper. Peak blood lactate increased in all subjects to the end of the first taper before declining by the end of the second training period, rising again to baseline levels during the second taper. The change in gamma2 was examined relative to the work rate achieved in cycle ergometry above an initial baseline score (deltaCP) and against concurrent peak blood lactate. There was a clear upward shift in gamma2 above baseline throughout the first and second training and taper in two subjects; this was less clear in the remaining two subjects, each of whom had a lower deltaCP. We conclude that this indicates improved lactate clearance, manifest by the change in gamma2 induced by endurance training. PMID- 10362392 TI - Measuring running speed using photocells. AB - Photocell timing systems are used routinely to measure running speeds. In this study, the accuracy of such systems was evaluated using centre of mass speed estimates from three-dimensional video analysis as criteria. One subject ran at five nominal speeds (5-9 m x s(-1)) for each of five separations (1.6-2.4 m) between consecutive photocells. Running speeds were calculated from the photocell data using single beam and double beam systems. For single beam systems, the start of the first break of a beam and the start of the longest break of a beam were used as trigger criteria. For double beam systems, the first occurrence of both beams being broken and the start of the longest double break were used as trigger criteria. Root mean square speed errors were smaller for the double beam systems. The longest break criterion gave smaller root mean square errors than the first break criterion. In general, errors in speed were smaller for greater photocell separations. An error of 0.1 m x s(-1) was achieved using a single beam system set at hip height with a longest break criterion for photocell separations of around two stride lengths. The advantage of using a double beam system is that it achieves this accuracy without the need to adjust photocell separation for different stride lengths. PMID- 10362393 TI - Bird's-eye view of nosocomial infections in medical ICU: blue bugs, fungi, and device-days. PMID- 10362394 TI - Use of nitric oxide synthase inhibitors to treat septic shock: the light has changed from yellow to red. PMID- 10362395 TI - Nitric oxide synthase inhibitors--a mechanism-based treatment of septic shock. PMID- 10362396 TI - Augmented enhancement of in vitro production of inflammatory cytokines... PMID- 10362397 TI - Polymerase chain reaction: a new chapter in critical care diagnosis. PMID- 10362398 TI - Capnography in critical care: accurate assessment of ARDS therapy? PMID- 10362399 TI - Moon phases and ocean tides: the relationship between the inspiratory-expiratory phases of mechanical ventilation and right ventricular function. PMID- 10362400 TI - ATP and sepsis. PMID- 10362401 TI - How low can you go? Blood pressure control after intracranial hemorrhage. PMID- 10362402 TI - Measurement inside the box. PMID- 10362403 TI - Low-dose inhaled nitric oxide and oxygenation in pediatric hypoxic respiratory failure. Wrong bullet, wrong target. PMID- 10362404 TI - Critical care medicine, terrorism and disasters: are we ready? PMID- 10362405 TI - Buffer treatment for cardiac resuscitation: putting the cart before the horse? PMID- 10362406 TI - Management of postoperative chylothorax with nitric oxide: a critical review. PMID- 10362407 TI - Arterial puncture during central venous catheter insertion. PMID- 10362409 TI - Nosocomial infections in medical intensive care units in the United States. National Nosocomial Infections Surveillance System. AB - OBJECTIVE: To describe the epidemiology of nosocomial infections in medical intensive care units (ICUs) in the United States. DESIGN: Analysis of ICU surveillance data collected through the National Nosocomial Infections Surveillance (NNIS) System between 1992 and 1997. SETTING: Medical ICUs in the United States. PATIENTS: A total of 181,993 patients. MEASUREMENTS AND MAIN RESULTS: Nosocomial infections were analyzed by infection site and pathogen distribution. Urinary tract infections were most frequent (31%), followed by pneumonia (27%) and primary bloodstream infections (19%). Eighty-seven percent of primary bloodstream infections were associated with central lines, 86% of nosocomial pneumonia was associated with mechanical ventilation, and 95% of urinary tract infections were associated with urinary catheters. Coagulase negative staphylococci (36%) were the most common bloodstream infection isolates, followed by enterococci (16%) and Staphylococcus aureus (13%). Twelve percent of bloodstream isolates were fungi. The most frequent isolates from pneumonia were Gram-negative aerobic organisms (64%). Pseudomonas aeruginosa (21%) was the most frequently isolated of these. S. aureus (20%) was isolated with similar frequency. Candida albicans was the most common single pathogen isolated from urine and made up just over half of the fungal isolates. Fungal urinary infections were associated with asymptomatic funguria rather than symptomatic urinary tract infections (p < .0001). Certain pathogens were associated with device use: coagulase-negative staphylococci with central lines, P. aeruginosa and Acinetobacter species with ventilators, and fungal infections with urinary catheters. Patient nosocomial infection rates for the major sites correlated strongly with device use. Device exposure was controlled for by calculating device-associated infection rates for bloodstream infections, pneumonia, and urinary tract infections by dividing the number of device-associated infections by the number of days of device use. There was no association between these device-associated infection rates and number of hospital beds, number of ICU beds, or length of stay. There is a considerable variation within the distribution of each of these infection rates. CONCLUSIONS: The distribution of sites of infection in medical ICUs differed from that previously reported in NNIS ICU surveillance studies, largely as a result of anticipated low rates of surgical site infections. Primary bloodstream infections, pneumonia, and urinary tract infections associated with invasive devices made up the great majority of nosocomial infections. Coagulase-negative staphylococci were more frequently associated with primary bloodstream infections than reported from NNIS ICUs of all types in the 1980s, and enterococci were a more frequent isolate from bloodstream infections than S. aureus. Fungal urinary tract infections, often asymptomatic and associated with catheter use, were considerably more frequent than previously reported. Invasive device-associated infections were associated with specific pathogens. Although device-associated site-specific infection rates are currently our most useful rates for performing comparisons between ICUs, the considerable variation in these rates between ICUs indicates the need for further risk adjustment. PMID- 10362408 TI - Increase in endotoxin-induced mucosal permeability is related to increased nitric oxide synthase activity using the Ussing chamber. AB - OBJECTIVE: To determine if nitric oxide production is associated with increased intestinal permeability after endotoxin challenge using the ex vivo Ussing chamber. SUBJECTS: Ileal mucosal membranes harvested from normal rats weighing 300 to 420 g. INTERVENTIONS: Endotoxin (lipopolysaccharide), 1, 10, 100 microg/ mL, or saline was placed on the serosal side of ileal mucosal membranes mounted in Ussing chambers after 10(9) Escherichia coli C-25 had been placed on the mucosal side of the ileal membranes (n = 6-7/group). In a second set of experiments, ileal membranes were exposed to 100 microg/mL lipopolysaccharide with or without the addition of the nitric oxide synthase inhibitor, N(G) monomethyl-L-arginine at a concentration of 10 mM (n = 7-8/group). MAIN OUTCOME MEASURE: Bacterial translocation of E. coli C-25 from the mucosal to the serosal side of the ileal membrane was measured every hour during the 3-hr experimental period, as were serial measurements of the potential difference and resistance values of the ileal membranes. At the conclusion of the 3-hr period, the ileal membranes were harvested and levels of inducible nitric oxide synthase and constitutive nitric oxide synthase activity were measured. RESULTS: The incidence of E. coli C-25 passage across the ileal membranes mounted in the Ussing chambers was significantly increased in the ileal membranes exposed to 10 or 100 microg/mL of lipopolysaccharide (71% and 86%, respectively) vs. the control membranes (0%) or the membranes exposed to 1 microg/mL of lipopolysaccharide (0%) (p < .05). This increase in E. coli C-25 passage in the ileal membranes exposed to 10 or 100 microg/mL of lipopolysaccharide was associated with a decrease in ileal membrane resistance and an increase in inducible nitric oxide synthase activity (p < .05). The addition of N(G)-monomethyl-L-arginine protected against lipopolysaccharide induced bacterial translocation and prevented the lipopolysaccharide-induced increase in ileal membrane inducible nitric oxide synthase activity. CONCLUSION: These results indicate that lipopolysaccharide induction of increased ileal inducible nitric oxide synthase activity is necessary for lipopolysaccharide induced E. coli C-25 translocation to occur in normal ileal mucosal membranes tested in the Ussing chamber system. PMID- 10362410 TI - Effects of epinephrine, norepinephrine, or the combination of norepinephrine and dobutamine on gastric mucosa in septic shock. AB - OBJECTIVES: To compare in the same patient with septic shock, respective effects of epinephrine, norepinephrine, and the combination of norepinephrine and dobutamine (5 microg/kg/min) on systemic hemodynamic parameters and gastric mucosal perfusion using gastric tonometry and laser-Doppler flowmetry techniques. DESIGN: Prospective, controlled, randomized, crossover study. SETTING: University hospital intensive care unit. PATIENTS: Twelve patients with septic shock. INTERVENTIONS: Each patient received in a random succession epinephrine, norepinephrine, and norepinephrine plus dobutamine. Dosages of epinephrine and norepinephrine were adjusted to achieve a mean arterial pressure between 70 and 80 mm Hg. A laser-Doppler probe and a tonometer were introduced into the gastric lumen. MEASUREMENTS AND MAIN RESULTS: The increase in gastric mucosal perfusion detected by laser-Doppler flowmetry was higher with epinephrine and the combination of norepinephrine and dobutamine than with norepinephrine alone (p < .05). In addition, the ratio of gastric mucosal perfusion (local oxygen delivery) to systemic oxygen delivery was increased after norepinephrine plus dobutamine as compared with norepinephrine alone and epinephrine (p< .05). Although values of intramucosal pH and gastroarterial PCO2 tended to be higher with norepinephrine plus dobutamine compared with those obtained with norepinephrine and epinephrine, differences were not statistically significant. CONCLUSIONS: For the same mean arterial pressure in patients with septic shock, our study showed that administration of epinephrine increased gastric mucosal perfusion more than norepinephrine administration alone. Addition of dobutamine (5 microg/kg/ min) to norepinephrine improved gastric mucosal perfusion. This result could be explained by a vasodilating effect of dobutamine on gastric mucosal microcirculation. PMID- 10362411 TI - Prognostic usefulness of scoring systems in critically ill patients with severe acute pancreatitis. AB - OBJECTIVE: To compare prognostic scoring systems in a retrospective series of patients with severe acute pancreatitis admitted to a surgical intensive care unit (ICU). METHOD: Between January 1992 and December 1996, the charts of all patients with a discharge code of acute pancreatitis were reviewed. There were 273 charts reviewed. Of these, 12 were admitted to the surgical ICU with a diagnosis of severe acute pancreatitis. A preliminary analysis of the data considers descriptive summary statistics, such as the mean and the range. The Spearman's rank-correlation test was computed to assess concordance between the following: a) length of stay and Ranson criteria; b) length of stay and Acute Physiology and Chronic Health Evaluation (APACHE) III score; and c) length of stay and modified Glasgow Coma score. Also, an unpaired t-test was used to obtain concordance between the following: a) death and Ranson; b) death and APACHE III; and c) death and modified Glasgow Coma score. RESULTS: The prognostic score for APACHE III, Ranson criteria, and modified Glasgow Coma score were compared with the patients' length of stay. Patients who had >5 Ranson criteria, modified Glasgow Coma scores of >4, and APACHE III scores of >30 at 96 hrs (mean 71+/-16 [SD]; p < .0) subsequently died. These two patients were excluded from the Spearman's rank-correlation tests. The mean length of stay in our sample was 61.8 (range, 7-201) days. The mean Ransom criteria was 4.3 (range, 1-9). The mean 96 hr APACHE III score was 33.3 (range, 0-83). The Spearman's rank-correlation between length of stay and Ranson criteria was 0.68, with a corresponding p value of .03. Similar results were observed for the length of stay and APACHE III at 96 hrs (correlation, 0.77; p = .0098) and the length of stay and the modified (correlation, 0.78; p = .007). These data reveal that the magnitude of correlation between the length of stay and the 96-hr APACHE III and modified Imrie is larger than that between length of stay and Ranson criteria. CONCLUSIONS: Once a patient is admitted to the surgical ICU, several predictors of mortality or complications that will require long hospitalization times are evident. In this sample of patients, APACHE III scores >30 at 96 hrs, 5 or more Ranson criteria, and a modified Imrie (Glasgow) score of >3 predicted those who died or had multiple complications. Those patients with combined 48-hr and 96-hr APACHE III scores of >60 either died or had hospitalizations of >60 days. These patients had major pancreatic complications that included pancreatic necrosis, pancreatic abscess, pseudocyst, hemorrhagic pancreatitis, and pancreatic ascites. PMID- 10362412 TI - Effect of the nitric oxide inhibitor, L-N(G)-monomethylarginine, on accumulation of interleukin-6 and interleukin-8, and nuclear factor-kappaB activity in a human endothelial cell line. AB - OBJECTIVE: To determine the effect of the nitric oxide synthase inhibitor, L-N(G) monomethylarginine, on interleukin-6 and interleukin-8 accumulation, and nuclear factor-kappaB expression in an endothelial cell model of sepsis. DESIGN: Controlled cell culture experiments examining the immunomodulatory effects of nitric oxide synthase inhibition. SUBJECTS: A human endothelial cell line (EA.hy926). MEASUREMENTS AND RESULTS: Cells were incubated with tumor necrosis factor-alpha and interleukin (IL)-1beta in the presence of L-N(G) monomethylarginine (L-NMMA). IL-6 and IL-8 were measured in culture supernatants using enzyme immunoassay. Nuclear factor-kappaB was measured using electrophoretic mobility shift assay and was quantified using phosphorimaging. IL 6 accumulation was decreased (p < .05) and IL-8 accumulation increased (p < .01) with L-NMMA. Increased nuclear factor-kappaB expression in stimulated cells was unaltered on exposure to L-NMMA. Cell viability was unaffected. CONCLUSIONS: Excessive production of nitric oxide has been implicated in septic shock, and the use of nitric oxide synthase inhibitors has been suggested. The immunoregulatory actions of nitric oxide synthase inhibitors affects the profile of cytokine release. This effect is not mediated through modulation of nuclear factor-kappaB. These findings have implications for the use of nitric oxide synthase inhibiting agents in septic shock. PMID- 10362413 TI - An open-label dose escalation study of the nitric oxide synthase inhibitor, N(G) methyl-L-arginine hydrochloride (546C88), in patients with septic shock. Glaxo Wellcome International Septic Shock Study Group. AB - OBJECTIVE: To assess the effects of the nitric oxide synthase inhibitor, 546C88, in patients with septic shock and to evaluate the range of dose rates that sustain mean arterial pressure (MAP) of > or =70 mmHg. DESIGN: Multicenter, open label, uncontrolled, dose range finding study. SETTING: Ten intensive care units in Europe and the United States. PATIENTS: Thirty-two patients with septic shock diagnosed within <24 hrs. INTERVENTIONS: Patients received a fixed dose rate of 546C88 at either 1(n = 6), 2.5 (n = 6), 5 (n = 4), 10 (n = 5), or 20 mg/kg/hr (n = 11) by intravenous infusion for up to 8 hrs. Conventional vasoactive therapy was restricted to norepinephrine and/or dobutamine. During 546C88 therapy, MAP was to be maintained between 70 and 90 mm Hg, while attempting to withdraw any concurrent norepinephrine. MEASUREMENTS AND MAIN RESULTS: Systemic and pulmonary hemodynamics, blood gases, and plasma nitrate were assessed. Infusion of 546C88 (1-20 mg/kg/hr) for up to 8 hrs enabled a 60 to 80% reduction of the norepinephrine dose rate in all cohorts, while MAP was sustained at >70 mm Hg. There was an increase in vascular tone and a decrease in cardiac index within the 1st hr of therapy. Systemic vascular resistance returned toward baseline with reduction of concomitant administration of norepinephrine. The decline in oxygen delivery was associated with an increase in extraction and, therefore, the maintenance of oxygen consumption. There was a sustained reduction of venous admixture within the 1st hr of therapy. 546C88 was not associated with any major or dose-dependent adverse effect on pulmonary, hepatic, or renal function. CONCLUSIONS: Treatment with the nitric oxide synthase inhibitor, 546C88, can restore the balance of vasomotor tone, thereby, maintaining blood pressure and reducing or eliminating the requirement for norepinephrine therapy in patients with septic shock. Infusion of 546C88 (1-20 mg/kg/hr) appears to have a satisfactory overall safety profile. PMID- 10362414 TI - Do the components of heat and moisture exchanger filters affect their humidifying efficacy and the incidence of nosocomial pneumonia? AB - OBJECTIVES: To compare the efficiency of two heat and moisture exchange filters (HMEFs) of different compositions of the humidifying capacity and the rate of bronchial colonization and ventilator-associated pneumonia in patients in the intensive care unit (ICU). DESIGN: Prospective, randomized study. SETTING: ICU of a university hospital. PATIENTS: All patients who required mechanical ventilation for 24 hrs or more during the study period. INTERVENTIONS: At admission to the ICU, patients were randomly assigned to one of two groups. In one group, the patients were ventilated with Humid-Vent Filter Light HMEF. The condensation surface was made of paper impregnated with CaCl2. The filter membrane was made of polypropylene. In the other group, the patients were ventilated with the Clear ThermAl HMEF (Intersurgical, France). The condensation surface was made of plastic foam impregnated with AlCl2. The filter membrane was made of two polymer fibers (modacrylic and polypropylene). In both groups, HMEFs were changed daily. MEASUREMENTS AND MAIN RESULTS: Seventy-seven patients were ventilated for 19+/-7 days with the Humid-Vent Filter Light HMEF and 63 patients for 17+/-6 days with the Clear ThermAl HMEF. Patients ventilated with the Humid-Vent Filter Light underwent 8.7+/-3.7 tracheal aspirations and 1.2+/-2.0 instillations per day and those with the Clear ThermAl, 8.2+/-3.9 and 1.5+/-2.4 per day, respectively (NS). The abundance of tracheal secretions and the presence of blood and viscosity, as evaluated by semiquantitative scales, were similar in both groups. One episode of tracheal tube occlusion was observed with the Humid-Vent Filter Light HMEF and none with the other HMEF (NS). Tracheal colonization was observed at a rate of 91% with the Humid-Vent Filter Light and 97% with the Clear ThermAl (NS). The rate of ventilator-associated pneumonia was similar in both groups (35%). Bacteria responsible for tracheal colonization and pneumonia were similar in both groups. CONCLUSIONS: Despite differences in their components, the two HMEFs that were tested achieved similar performances in terms of humidification and heating of inspired gases. Only one episode of endotracheal tube occlusion was detected, and very few patients (three in each group) had to be switched to an active heated humidifier. No difference was observed either in the rate of tracheal colonization or of ventilator-associated pneumonia. These data show that the Humid-Vent Filter Light and the Clear ThermAl HMEFs are suited for use with ICU patients. PMID- 10362415 TI - Augmented enhancement of in vitro production of inflammatory cytokines in peripheral blood mononuclear cells in patients undergoing simultaneous resection of the liver and gastrointestinal tract. AB - OBJECTIVE: To determine changes in the production of inflammatory cytokines and acute-phase proteins, and in the priming of peripheral blood mononuclear cells (PBMC), as mechanisms for the high incidence of postoperative complications in patients who have undergone hepatectomy simultaneously with resection of the gastrointestinal tract. DESIGN: Prospective, clinical study for 3 wks after operation. SETTING: A surgical department in a university hospital. PATIENTS: Twenty-one consecutive adult patients with synchronous and metachronous hepatic metastases from gastrointestinal malignancies, curatively resected by simultaneous resection (group A, n = 9) or by hepatectomy alone (group B, n = 12), and 15 patients with gastrointestinal malignancies undergoing curative resection (group C). INTERVENTION: Peripheral venous blood samples collected before operation and on days 1, 3, 5, 7, 10, 14, and 21 after operation. MEASUREMENTS AND MAIN RESULTS: The serum and plasma levels of acute-phase proteins, interleukin (IL)-1beta, IL-6, tumor necrosis factor (TNF)-alpha, and endotoxin were measured. The in vitro production of IL-1beta and TNF-alpha by PBMC was also determined by the stimulation of lipopolysaccharide. The incidence of postoperative complications was significantly higher in group A than in groups B and C. The serum levels of IL-6 increased significantly, with a peak at postoperative day 1 in all groups, and the peak levels of IL-6 in groups A and B were significantly higher than that in group C. The serum levels of all acute phase proteins measured in this study (alpha1-antitrypsin, haptoglobin, and C reactive protein) increased markedly after operation in group C (p < .05). In group A, only C-reactive protein increased after operation, but its peak level was lower than in groups B and C (p < .05). Although IL-1beta and TNF-alpha in the serum were not detectable in any of the groups during the entire study period, the lipopolysaccharide-induced in vitro production of IL-1beta and TNF alpha by PBMC in all groups was significantly elevated after operation, with a peak at days 1 and 3 after operation, respectively. In addition, the elevation of the in vitro production of IL-1beta and TNF-alpha in group A was significantly greater than that in group C, lasting until postoperative day 5 (IL-1beta) and postoperative day 10 (TNF-alpha). The levels of plasma endotoxin increased significantly in all groups, with a peak at day 1 after operation, and the peak levels were significantly higher in group A than in groups B and C. There was a significant correlation between the peak levels of in vitro TNF-alpha production and the peak levels of plasma endotoxin (r2 = .331, p< .01). CONCLUSIONS: The augmented enhancement of the priming of PBMC as a result of surgery in patients undergoing simultaneous resection of the liver and gastrointestinal tract, together with the reduced synthesis of the acute-phase reactants and impaired host defense mechanisms, might be responsible for the high incidence of postoperative complications, possibly because subsequent exposure of primed macrophages/monocytes to triggering substances such as endotoxin and bacterial components after operation results in inappropriate production of inflammatory cytokines. PMID- 10362416 TI - The use of polymerase chain reaction to detect septicemia in critically ill patients. AB - OBJECTIVE: To describe the use of bacterial DNA amplification of conserved bacterial 16S ribosomal DNA nucleotide sequences by polymerase chain reaction (PCR) to detect the presence of septicemia in critically ill septic patients. DESIGN: Case series of blood samples from septic patients comparing the PCR results with conventional blood culture results. SETTING: A general intensive care unit in a tertiary referral hospital. PATIENTS: Two sets of samples (n = 101 and n = 55) from patients diagnosed as clinically septic and requiring blood cultures. They were classified by internationally accepted criteria into systemic inflammatory response syndrome, severe sepsis, and septic shock groups. INTERVENTIONS: Blood samples taken in a sterile fashion concurrently for blood culture, and PCR of the bacterial 16S ribosomal RNA gene in leukocytes and plasma. Two different DNA extraction techniques for PCR were tried sequentially. MEASUREMENTS AND MAIN RESULTS: Blood culture and PCR positivity were measured in relation to the clinical classification of severity of sepsis. Using the initial extraction method (n = 101), ten patients were positive by both PCR and blood culture, eight patients were PCR positive and blood culture negative, and seven patients were blood culture positive and PCR negative. From the clinical criteria, PCR detected at least six true positives that had been missed on blood culture and missed four true Gram-positive bacteremias. When the initial code was broken, this deficiency was rectified using the improved extraction technique (n = 55), in which ten patients were positive by PCR and blood culture, 29 patients were PCR positive and blood culture negative, and two patients were PCR negative and PCR positive. CONCLUSIONS: We conclude that the use of PCR (for the 16S ribosomal DNA in the plasma) was significantly more sensitive than the use of conventional blood culturing techniques for the detection of bacteremia in seriously ill patients. This could prove to be a valuable adjunct to conventional blood cultures. PMID- 10362417 TI - Prolonged partial liquid ventilation in spontaneously breathing awake animals. AB - OBJECTIVE: To date, studies of partial liquid ventilation (PLV) have examined its effects acutely in anesthetized and mechanically ventilated subjects. We set out to develop a model of prolonged PLV in awake, spontaneously breathing animals. DESIGN: Animal case series SETTING: Cardiopulmonary physiology laboratory. SUBJECTS: Fifteen New Zealand white rabbits (3.2+/-0.39 kg). INTERVENTIONS: Animals were anesthetized and instrumented with a novel technique allowing percutaneously assisted placement of an intratracheal catheter with a subcutaneously tunneled externalized free end. After anesthetic recovery, PLV was performed in spontaneously breathing unsedated animals. MEASUREMENTS AND MAIN RESULTS: Evaporative losses were determined using a single 10 mL/kg perflubron dose (n = 5); hourly radiographs were obtained until residual perflubron was minimal. For prolonged PLV (n = 10), a 10-mL/kg initial perflubron dose was followed every 4 hrs with 5-mL/kg supplements. Radiographs were obtained immediately before and after perflubron administration and were scored (0-5) by a radiologist blinded to dosing regimen and time interval. Data were analyzed with ANOVA and Student's t-test with correction for multiple comparisons. Initial filling was nearly complete (score = 4.8+/-0.42); lungs were maintained approximately half-filled through 4 hrs (score = 2.53+/-0.71). By 6 hrs, the majority of perflubron had evaporated (score = 1.75+/-0.53). Over 24 hrs, radiographs documented continuous perflubron exposure (postffill = 4.53+/-0.64, prefill = 3.40+/-0.71, average = 3.97+/-0.43); scores were comparatively higher after filling (after = 4.53+/-0.64, before = 3.4+/-0.71, p< .001). Left and right lung volumes were equivalent (left = 4.06+/-0.47, right = 3.89+/-0.39, p = .027). All animals survived the 24 hrs of PLV. CONCLUSIONS: Percutaneously assisted intratracheal cannulation with catheter exteriorization permits prolonged PLV in spontaneously breathing, unsedated animals. Continuous perfluorocarbon exposure with this method is reproducible, consistent, and well tolerated for 24 hrs in uninjured animals. PMID- 10362418 TI - Utility of deadspace and capnometry measurements in determination of surfactant efficacy in surfactant-depleted lungs. AB - OBJECTIVE: To investigate if bronchoalveolar lavage leads to increased alveolar and physiologic deadspace or a deadspace/ tidal volume ratio and if surfactant replacement restores the lung to its prelavage condition. DESIGN: Prospective, animal cohort study. SETTING: An animal laboratory in a university medical center. SUBJECTS: Seven adult rabbits receiving artificial ventilation. METHODS: Our single-breath CO2 analysis station contained the following equipment: pneumotachometer Ventrak 1550, a mainstream capnometer Capnogard 1265, a signal processor, and computer software. Repeated bronchoalveolar lavage was performed in seven adult rabbits to simulate acute respiratory distress syndrome. Surfactant therapy was administered after bronchoalveolar lavage induced a 20% reduction in baseline arterial PO2. The calculated parameters of alveolar and physiologic deadspace and the deadspace/tidal volume ratio were derived from the single-breath CO2 plot by Ventrak 1550 in combination with the Capnogard 1265. The arterial end-tidal Pco2 difference, the alveolar-arterial PO2 difference, and the arterial/alveolar PO2 ratio were obtained by capnography and arterial blood gas analysis. Measurements of these parameters were performed before bronchoalveolar lavage, during bronchoalveolar lavage, and after surfactant application. MEASUREMENTS AND MAIN RESULTS: The alveolar and physiologic deadspace and the deadspace/tidal volume ratio were significantly higher in lavaged animals. After application of natural surfactant, these parameters were significantly reduced but the baseline values could not be reached. Bronchoalveolar lavage led to a significant fall in the arterial/alveolar PO2 ratio, which increased after surfactant therapy. There was a negative correlation between the arterial/alveolar PO2 ratio and the deadspace/tidal volume ratio. The alveolar and physiologic deadspace and the deadspace/tidal volume ratio correlated with the arterial end-tidal Pco2 difference. The best correlation was obtained between the arterial end-tidal Pco2 difference and the alveolar deadspace/tidal volume ratio (r = 0.98). CONCLUSIONS: Bronchoalveolar lavage elevates the alveolar and physiologic deadspace and the deadspace/tidal volume ratios and is combined with a fall in the arterial/alveolar PO2 ratio. Surfactant treatment improves the gas exchange but does not restore the lung to its prebronchoalveolar lavage condition, which indicates that the exogenous surfactant affects only partly the recruitment of the atelectatic areas. PMID- 10362419 TI - Phase-related changes in right ventricular cardiac output under volume-controlled mechanical ventilation with positive end-expiratory pressure. AB - OBJECTIVE: To examine determinants of right ventricular function throughout the ventilatory cycle under volume-controlled mechanical ventilation with various positive end-expiratory pressure (PEEP) stages. DESIGN: Prospective observational animal pilot study. SETTING: Animal research laboratory at a university hospital. SUBJECTS: Eight healthy swine under volume- controlled mechanical ventilation. INTERVENTIONS: Flow probes were implanted in eight swine in order to continuously measure blood flow in the pulmonary artery and inferior vena cava. After a recovery phase of 14 days, the swine were subjected to various PEEP stages (0, 5, 10 cm H2O) during volume-controlled positive pressure ventilation. MEASUREMENTS AND MAIN RESULTS: Continuous flow measurement took place in the pulmonary artery and inferior vena cava. Data on standard hemodynamic parameters were additionally acquired. Respiration-phase-specific analysis of right ventricular cardiac output and of additional hemodynamic function parameters followed, after calculation of mean values throughout five respiration cycles. PEEP at 5 cm H2O led to significant decreases in inferior vena cava flow (4.1%), and in right ventricular cardiac output (5.2%); the respective decreases at PEEP 10 cm H2O were 13.9% and 18.3%. In the inspiration phase at PEEP 10 cm H2O, results revealed an overproportionally pronounced decrease in comparison with the expiration phase in inferior vena cava flow (-24.6% vs. -10%) and right ventricular cardiac output ( 35% vs. -13.5%). This phenomenon is presumably caused by a PEEP-related increase in mean airway pressure by the amount of 10.7 cm H2O in inspiration. CONCLUSIONS: Increases in PEEP during volume-controlled mechanical ventilation leads to respiration-phase-specific reduction of right ventricular cardiac output, with a significantly pronounced decrease during the inspiration phase. This decrease in cardiac output should be taken into particular consideration for patients with already critically reduced cardiac output. PMID- 10362420 TI - Salutary effects of ATP-MgCl2 on the depressed endothelium-dependent relaxation during hyperdynamic sepsis. AB - OBJECTIVES: Studies have shown that endothelium-dependent relaxation (mediated by endothelium-derived nitric oxide) is depressed during the early, hyperdynamic stage of sepsis. Although it is known that ATP-MgCl2 produces beneficial effects following various adverse circulatory conditions, it remains unknown whether this agent attenuates the depressed endothelium-dependent relaxation during early sepsis. The aim of this study, therefore, was to determine whether or not the administration of ATP-MgCl2 early after the onset of sepsis improves or maintains endothelium-dependent relaxation. DESIGN: Prospective, controlled animals study. SETTING: A university research laboratory. SUBJECTS: Adult male Sprague-Dawley rats were subjected to polymicrobial sepsis by cecal ligation and puncture (CLP), followed by administration of 3 mL/100 g body weight normal saline to these and sham-operated rats. INTERVENTIONS: At 1 hr after CLP, ATP-MgCl2 (50 micromol/kg body weight) or an equivalent volume of normal saline was infused intravenously over 90 mins. MEASUREMENTS AND MAIN RESULTS: At 5 hrs or 10 hrs after CLP (i.e., the early, hyperdynamic stage of sepsis), the thoracic aorta was isolated, cut into rings, and placed in organ chambers. Norepinephrine was used to preconstrict vessel rings. Dose responses for an endothelium-dependent vasodilator, acetylcholine (ACh, via endothelium-dependent nitric oxide), and an endothelium independent vasodilator, nitroglycerin, were determined. These results indicate that endothelium-dependent relaxation induced by ACh was significantly depressed at 5 and 10 hrs after CLP. Administration of ATP-MgCl2 after the onset of sepsis, however, maintained ACh-induced vascular relaxation. In contrast, no significant difference in nitroglycerin-induced vascular relaxation as well as norepinephrine induced contraction was observed, irrespective of administration of ATP-MgCl2. CONCLUSION: Since administration of ATP-MgCl2 prevents the impaired vascular relaxation to the endothelium-dependent vasodilator ACh, this agent may be a useful adjunct for maintaining endothelial cell function during the hyperdynamic stage of sepsis. PMID- 10362421 TI - Pharmacologic reduction of mean arterial pressure does not adversely affect regional cerebral blood flow and intracranial pressure in experimental intracerebral hemorrhage. AB - OBJECTIVE: To determine the effect of mean arterial pressure (MAP) reduction on regional cerebral blood flow and intracranial pressure (ICP) in intracerebral hemorrhage. We tested the hypothesis that there is ischemia in the perihematoma region after intracerebral hemorrhage, which can be exacerbated by a pharmacologic reduction of MAP. DESIGN: Prospective, controlled, laboratory trial. SETTING: Animal research laboratory. SUBJECTS: Eighteen mongrel dogs, weighing 15 to 25 kg. INTERVENTIONS: We introduced intracerebral hemorrhage in 12 anesthetized dogs by autologous blood injection under arterial pressure in the deep white matter adjacent to the left caudate region. We measured serial regional cerebral blood flow using radiolabeled microspheres in animals with two different volumes of injected blood (2.8 mL [Group A, n = 6] and 4.4 mL [Group B, n = 6]) and compared them with control animals (n = 6). Intravenous labetalol was administered 90 mins after administration of hematoma, while maintaining cerebral perfusion pressure >65 mm Hg. Regional cerebral blood flow measurements were repeated 10 and 30 mins after labetalol administration. MAP and ICP were monitored continuously using intra-arterial and cisterna magna catheters, respectively. MEASUREMENTS AND MAIN RESULTS: Compared with control animals, significant elevation in ICP was observed in Groups A and B and elevation in MAP was observed in Group B at 45 mins after injection of blood. These hemodynamic alterations were not accompanied by any significant differences in regional cerebral blood flow in any group. Administration of labetalol resulted in a decrease in MAP (mm Hg+/-SEM) in Groups A (119.0+/-9.2 to 103.0+/-9.1) and B (124.5+/-7.4 to 100.5+/-4.8) and controls (103.5+/-4.3 to 85.0+/-8.0). No differences were observed in regional cerebral blood flow after MAP reduction in both Groups A and B compared with controls in regions around or distant to the hematoma. There were no changes in ICP in Groups A and B both at 10 and 30 mins after reduction in MAP compared with pretreatment values. CONCLUSIONS: In our model, pharmacologic reduction of MAP within the normal autoregulatory limits of cerebral perfusion pressure, 90 mins after onset, had no adverse effect on ICP and regional cerebral blood flow in regions around or distant to the hematoma. These results support the controlled use of antihypertensive treatment in intracerebral hemorrhage in the initial time period. PMID- 10362422 TI - Subarachnoid hemorrhage in rats: effect of singular or sustained hemodilution with alpha-alpha diaspirin crosslinked hemoglobin on cerebral hypoperfusion. AB - OBJECTIVE: To evaluate the effect of singular or sustained hemodilution, with alpha-alpha diaspirin crosslinked hemoglobin (DCLHb), on the area of hypoperfusion after subarachnoid hemorrhage. DESIGN: Prospective animal study. SETTING: Animal research laboratory. SUBJECTS: Isoflurane anesthetized, mechanically ventilated rats. INTERVENTIONS: Subarachnoid hemorrhage was induced by injecting 0.3 mL of blood into the cisterna magna. The animals were randomly assigned to one of the following groups (n = 16 in each hemodilution group; eight animals received a single treatment of hemodilution after subarachnoid hemorrhage; and, for eight animals, treatment was sustained for 48 hrs): control group (n = 8), no hematocrit (45%) manipulation; DCLHb group (n = 16), hematocrit decreased to 30% with DCLHb; or Alb group (n = 16), hematocrit decreased to 30% with human serum albumin. After 48 hrs, the area of hypoperfusion (cerebral blood flow < 40 ml/100g/min) was determined with 14C-iodoantipyrine in five coronal brain sections. MEASUREMENTS AND MAIN RESULTS: For both singular and sustained treatment, the area of hypoperfusion was less in both hemodilution groups than in the control group (p<.05). For four of the five coronal brain sections, no differences were found between the DCLHb and Alb groups within a given hemodilution protocol. In addition, in four of the five coronal brain sections for the DCLHb hemodilution groups and in all five sections for the albumin hemodilution groups, the area of hypoperfusion was less for rats that received sustained hemodilution compared with their respective groups in the singular treatment protocol (p<.05). CONCLUSIONS: These data support the hypothesis that hemodilution with molecular hemoglobin decreases hypoperfusion after subarachnoid hemorrhage and that sustained hemodilution is more effective than singular treatment. The data do not support the notion that intravascular DCLHb has an adverse effect on cerebral ischemia after subarachnoid hemorrhage. PMID- 10362423 TI - Brain eigenfrequency shifting as a sensitive index of cerebral compliance in an experimental model of epidural hematoma in the rabbit: preliminary study. AB - OBJECTIVE: To verify brain eigenfrequency shifting after the occurrence of a lesion producing mass effect into the cranial vault. DESIGN: Experimental animal study. SETTING: Laboratory of experimental surgery affiliated with a university critical care department. SUBJECTS: Six adult male New Zealand white rabbits. INTERVENTIONS: A Camino ICP monitor was placed in the parenchyma, and a 5-Fr balloon-tipped catheter and accelerometer were placed into the epidural space. MEASUREMENTS: Before and after the introduction of successive 0.1-mL increments of autologous blood into the balloon, intracranial pressure (ICP) was recorded along with the accelerometer signal obtained during free vibration of the skull triggered by a calibrated hammer. Fast Fourier transformation of the digitized signal provided the eigenfrequency spectrum. The eigenfrequency showing the sharpest decrease after the initial 0.1-mL volume addition was considered as the best frequency, and its variation in response to subsequent 0.1-mL increments represents the brain eigenfrequency shifting. MAIN RESULTS: Brain eigenfrequency shifting to lower values occurs for small blood volume increments (up to 0.2 mL). When volume addition becomes >0.3 mL, brain eigenfrequency shifting to higher values is exhibited. The decrease in best frequency after the initial introduction of 0.1 mL is statistically significant (p = .003), in a range of volume in which no significant intracranial pressure difference appears. The respective variation of ICP is explained using a quadratic curve. For volumes of 0 to 0.1 mL, the change in ICP is not statistically significant (p = .08). CONCLUSIONS: Changes of the brain's physical characteristics by mass addition in the cranial vault can be expressed by brain eigenfrequency shifting. The method seems advantageous because it reliably detects mass additions at low levels where no ICP change occurs. Additionally, it provides serial measurements, and it is less invasive than the currently used methods for intracranial compliance. PMID- 10362424 TI - The Pediatric Risk of Mortality (PRISM) Score and Injury Severity Score (ISS) for predicting resource utilization and outcome of intensive care in pediatric trauma. AB - OBJECTIVE: Mortality prediction in trauma is assessed using the Injury Severity Score (ISS) and Revised Trauma Score using Trauma Injury Severity Score (TRISS) methodology. The Pediatric Risk of Mortality (PRISM) score assesses mortality risk in critically ill children. We compared the ability of PRISM and ISS (using TRISS methodology) to predict resource utilization and outcome in pediatric trauma. DESIGN: Retrospective chart and database review. SETTING: Pediatric intensive care unit (PICU). PATIENTS: Consecutive admissions to a PICU over a 2 yr period. MEASUREMENTS AND MAIN RESULTS: Demographic data including PICU resource utilization and outcome were recorded. Data were recorded on 1,052 admissions (31 deaths), including 125 pediatric trauma patients (11 deaths). Patients were stratified into low- and high-risk categories based on PRISM and ISS scores. Patients with PRISM scores <6 and ISS scores <10 were classified as low risk. While both low-risk PRISM and ISS scores readily identified survivors, PRISM was the more sensitive indicator of resource utilization. PRISM, however, performed less well in determining risk-adjusted mortality as compared with ISS. CONCLUSION: PRISM readily stratifies pediatric trauma patients for resource utilization. PRISM appears to underestimate mortality in pediatric trauma as compared with ISS using TRISS methodology. PMID- 10362426 TI - Molecular biology for the critical care physician. Part II: where are we now? AB - The extraordinary technical developments in molecular biology are having a profound impact in clinical medicine. The contribution of recombinant DNA technology in defining the molecular pathology of common disorders and of diagnostic molecular techniques for detection of infectious organisms are used as examples to demonstrate the clinical relevance of these developments. Finally, the potential use of DNA as a therapeutic drug (gene therapy) is addressed. PMID- 10362425 TI - Low-dose inhaled nitric oxide improves the oxygenation and ventilation of infants and children with acute, hypoxemic respiratory failure. AB - OBJECTIVE: To describe the effects of inhaled nitric oxide on oxygenation and ventilation in patients with acute, hypoxic respiratory failure and to characterize those who respond to low doses with a significant improvement in PaO2. DESIGN: Prospective dose response trial of inhaled nitric oxide. Patients who demonstrated a > or =15% improvement in PaO2 were randomized to receive conventional mechanical ventilation with or without prolonged inhaled nitric oxide. SETTING: Pediatric intensive care unit of a tertiary care children's hospital serving as a regional referral center for respiratory failure. PATIENTS: Pediatric patients with an acute parenchymal lung disease requiring mechanical ventilation, an F(IO2) of > or =0.5, a positive end-expiratory pressure of > or =7 cm H2O, and whose PaO2/FIO2 ratio was < or =160. INTERVENTIONS: PaO2, PaCO2, pH, heart rate, blood pressure, and methemoglobin were recorded at baseline and after inhaling 1, 5, 10, and 20 ppm of nitric oxide. Peak expiratory flow rate and mean airway resistance were measured while subjects received 0 and 20 ppm of inhaled nitric oxide. Patients were followed up until extubation or death. MEASUREMENTS AND MAIN RESULTS: Twenty-six patients (median age, 2.6 yrs [range, 1 mo-18.2 yrs]) were enrolled in the study. PaO2 increased (p< .001) and Pa(CO2) fell (p< .0001) from baseline with the administration of inhaled nitric oxide. There was no statistical difference among 1, 5, 10, and 20 ppm with regard to effects on oxygenation. Sixteen patients (62%) responded to inhaled nitric oxide with a > or =15% improvement in PaO2; 14 of these responses occurred at a dose of 1 or 5 ppm. Response to inhaled nitric oxide was not associated with age, length of intubation, presence of primary lung disease, chest radiograph, or illness severity. Among patients weighing < or =20 kg, responders showed a greater fall in mean airway resistance (p < .05) than nonresponders. Mortality was not influenced by prolonged inhaled nitric oxide when analyzed by intention to treat. Patients receiving prolonged inhaled nitric oxide at doses of < or =20 ppm maintained methemoglobin levels of <3.0% and circuit concentrations of NO2 of <1 ppm. CONCLUSIONS: Inhaled nitric oxide at doses of < or =5 ppm improves the oxygenation and (to a lesser extent) ventilation of most children with acute, hypoxic respiratory failure. The unpredictable response of patients necessitates individualized dosing of inhaled nitric oxide, starting at concentrations of < or =1 ppm. Inhaled nitric oxide at < or =20 ppm may exert a small salutary effect on bronchial tone. The benefits of prolonged inhaled nitric oxide remain unknown. PMID- 10362427 TI - Preparing hospitals for toxicological mass casualties events. AB - OBJECTIVE: For most hospital staffs, treatment of chemical casualties presents an obscure and even frightening situation. We report our unique experience from hospital drills in order to improve hospital preparedness for patient management under mass casualty conditions involving hazardous chemicals. SETTING: Twenty-one major hospitals in Israel. INTERVENTIONS: A unique hospital deployment plan for the management of chemical casualties was developed, and hospitals were required to have a full chemical practice drill every 3 to 5 yrs. These drills were designed as realistically as possible, and all included the use of personal protective equipment, decontamination, and treatment of simulated patients. Twenty-five percent of these patients, simulating children and adults, required intensive care and ventilation support. Hospitals were inspected and reviewed on the quality of treatment given and the overall continuity of care as well as on their administrative performance. RESULTS: Between 1986 to 1994, 30 full chemical practice drills were conducted in 21 major hospitals. Each drill included treatment of 100 to 400 simulated patients. The lessons from the hospital drills are described and were incorporated in the proposed revised hospital deployment plan. All hospitals significantly improved their ability to respond appropriately to these incidents. CONCLUSIONS: The level of preparedness for a chemical mass casualty scenario should be established according to the existing threat and the available resources. The proposed plan can serve as a basis for hospital planning and staff training worldwide, thus facilitating optimal care in the event of an incident involving toxic chemicals. A cost-effective scale for hospital preparation levels according to the existing threat is suggested. PMID- 10362428 TI - Tribonat--a comprehensive summary of its properties. AB - OBJECTIVE: To review available investigations describing the properties of the buffer mixture Tribonat. DATA SOURCES: Original reports published in peer reviewed medical journals. STUDY SELECTION: Review of 76 citations, including four original studies on the effect of Tribonat performed by or supervised by the author, and six original studies concerning Tribonat originating from the institution to which the author is affiliated. DATA EXTRACTION: Computer search of the literature regarding treatment with alkaline buffers during cardiopulmonary resuscitation. DATA SYNTHESIS: Routine buffering of acidosis has been questioned, but clinical situations still exist where such treatment is regarded as indicated. In such cases, a buffer with advantageous qualities and few side-effects is desirable. The hitherto commonly used buffers do not always fulfill these requirements, and a more profound knowledge of the alternative Tribonat may therefore be warranted. CONCLUSIONS: The reviewed articles support the assumption that Tribonat may offer important advantages over previously used buffers in situations where administration of an alkalinizing agent is indicated. PMID- 10362429 TI - An acute inflammatory response to the use of granulocyte colony-stimulating factor to prevent infections in patients with brain injury: what about the brain? PMID- 10362430 TI - Counterregulatory control of the acute inflammatory response: granulocyte colony stimulating factor has anti-inflammatory properties. PMID- 10362431 TI - Management of postoperative chylothorax with nitric oxide: a case report. AB - OBJECTIVE: To describe the use of inhaled nitric oxide in the management of refractory postoperative chylothorax. DESIGN: Case report. SETTING: A pediatric intensive care unit of a tertiary care children's hospital. PATIENT: A neonate with refractory chylothoraces complicated by moderate pulmonary hypertension after a complicated arterial switch operation. INTERVENTIONS: Administration of inhaled nitric oxide through a ventilator circuit. MEASUREMENTS AND MAIN RESULTS: The institution of inhaled nitric oxide at 20 ppm resulted in a marked reduction in chest tube drainage and a decrease in echocardiographically estimated pulmonary artery pressure from 50%-75% systemic to 30%-50% systemic. Chest tube drainage doubled when the nitric oxide was decreased to 10 ppm and, again, dramatically decreased after raising nitric oxide back to 20 ppm. After 8 days of nitric oxide therapy, the chest tube drainage ceased. Nitric oxide therapy was successfully discontinued 19 days after initiation, with no recurrence of chylothorax. There was no effect of nitric oxide on systemic blood pressure. Methemoglobin levels while on NO remained <1.7%. CONCLUSION: Consideration may be given to the use of inhaled nitric oxide in the therapy of refractory chylothoraces complicated by central venous hypertension. PMID- 10362432 TI - Central venous access: accidental arterial puncture in a patient with right-sided aortic arch. AB - OBJECTIVE: To describe an unusual case of accidental insertion of a central line into an anomalous right-sided aortic arch. DESIGN: Case report, clinical. SETTINGS: Community hospital, university-affiliated. CONCLUSIONS: Intraoperative radioscopy, chest radiographs, and pressure transducer monitoring usually allow for the prompt recognition of the accidental insertion of venous catheters into the arterial system. However, in the presence of a right-sided aortic arch, a central line could be erroneously inserted into the arterial system and the radiologic findings can give the false impression of a correct placement in the superior vena cava. PMID- 10362433 TI - Mergers and acquisitions among psychotropics: antidepressant takeover of anxiety may now be complete. PMID- 10362434 TI - Medication treatment for the severely and persistently mentally ill: the Texas Medication Algorithm Project. AB - This article provides an overview of the issues involved in developing, using, and evaluating specific medication guidelines for patients with psychiatric disorders. The potential advantages and disadvantages, as well as the essential elements in the structure of algorithms, are illustrated by experience to date with the Texas Medication Algorithm Project, a public-academic collaboration. Phase 1 entailed assembling research findings on the efficacy of medications for schizophrenic, bipolar, and major depressive disorders. This knowledge was evaluated for its quality and relevance, integrated with expert clinical judgment as well as input by practicing clinicians, family advocates, and patients. Phase 1 (the design and development of the algorithms) was followed by a feasibility test (Phase 2). Phase 3 is an ongoing evaluation comparing the clinical and economic effects of using specific medication guidelines (algorithms) versus treatment as usual in public sector patients with severe and persistent mental illnesses. PMID- 10362435 TI - Quetiapine, a novel antipsychotic: experience in elderly patients with psychotic disorders. Seroquel Trial 48 Study Group. AB - BACKGROUND: This uncontrolled trial examines the safety and effects of quetiapine, a new atypical antipsychotic, in elderly patients with psychotic disorders. METHOD: This is an ongoing, multicenter, open-label, 52-week trial of quetiapine in men and women at least 65 years old with DSM-IV psychotic disorders. Patients received quetiapine, 25 to 800 mg/day. Assessments included the 18-item Brief Psychiatric Rating Scale (BPRS), the Clinical Global Impressions scale (CGI), the Simpson-Angus Neurologic Rating Scale, and the Abnormal Involuntary Movement Scale (AIMS). RESULTS: An interim analysis was performed at 12 weeks with results from 151 patients. The median total daily dose was 100 mg/day. The most common adverse events were somnolence (32%), dizziness (14%), postural hypotension (13%), and agitation (11%). Extrapyramidal symptom adverse events occurred in 6% of patients. Mean Simpson-Angus Scale total score showed significant (p<.0001) improvement at endpoint; there were no changes in AIMS scores. BPRS total and CGI-Severity of Illness scores showed significant (p<.0001 and p<.01, respectively) improvement at endpoint. No clinically important effects were reported for hematologic or liver function test variables; small changes in mean free levorotatory thyroxine (T4) levels were not associated with substantial changes in mean thyroid-stimulating hormone concentration. Mean corrected QT interval (QTc) was unchanged, but a slight increase in mean heart rate was noted. CONCLUSION: Quetiapine was well tolerated in a nonrandomized study of elderly patients and was associated with improvement in symptoms. PMID- 10362436 TI - Once-weekly dosing of fluoxetine in the maintenance of remission in panic disorder. AB - BACKGROUND: Fluoxetine and its active metabolite norfluoxetine have long half lives of 4-6 days and 4-16 days, respectively. We postulated that, owing to the long elimination half-life, patients diagnosed with panic disorder might be maintained on fluoxetine taken once a week, after being treated initially with daily doses of fluoxetine. METHOD: Ten patients with DSM-III-R panic disorder were treated openly with fluoxetine, 20-40 mg daily. Once panic free, these patients were switched to once-weekly dosing of fluoxetine, and dosage was titrated as needed. RESULTS: All 10 patients successfully switched to once-weekly dosing. One patient reported recurrence of panic attacks 18 months after the switch. After a brief treatment for 4 weeks with benzodiazepines and daily fluoxetine, the patient was once again maintained on once-weekly dosing when rechallenged. Patients have been maintained in a panic-free state for up to 26 months with a single weekly dose of fluoxetine ranging from 10 to 60 mg. The medication was well tolerated. CONCLUSION: Fluoxetine at doses ranging from 10 to 60 mg administered once weekly appears to be effective maintenance treatment for patients with panic disorder who were initially treated successfully with daily fluoxetine. A once-weekly regimen may allow for considerable cost savings and may serve as a convenient alternative method for treating panic disorder. PMID- 10362437 TI - Study of impulse-control disorders among alcohol-dependent patients. AB - BACKGROUND: Impulse-control disorders (ICDs) include intermittent explosive disorder, kleptomania, trichotillomania, pyromania, and pathological gambling. Several studies have suggested that the incidence of pathological gambling is substantially higher in alcoholics than in the general population. The rate of co occurrence of other ICDs and alcohol dependence has never been systematically investigated. In our study, we assessed the frequency of all ICDs in a population of alcohol-dependent patients. We also examined the possibility that the presence of an ICD can correspond to earlier onset and more severe forms of alcoholism, which have a greater association with antisocial personality. METHOD: All patients hospitalized at our psychiatric unit for detoxification between January and August 1997 met DSM-IV criteria for alcohol dependence and were included in this study. Diagnosis of alcohol dependence was confirmed with the Mini International Neuropsychiatric Interview. ICDs were investigated using the Minnesota Impulsive Disorders Interview. All patients completed the Michigan Alcoholism Screening Test. RESULTS: Among the 79 patients included in the study, 30 (38.0%) met criteria for an ICD. Included in the study were 19 cases of intermittent explosive disorder, 7 cases of pathological gambling, 3 cases of kleptomania, and 1 case of trichotillomania. Patients with co-occurring ICDs were significantly younger than patients without an ICD (mean age = 40.7 vs. 44.5 years; p = .03). Patients with co-occurring pathological gambling were significantly younger at the onset of alcohol dependence than patients without ICDs (mean age = 19.5 vs. 25.9 years; p = .0008). Pathological gamblers had significantly longer duration of alcohol dependence compared with patients without ICDs (26.0 vs. 17.9 years; p = .02). Patients with co-occurring intermittent explosive disorder had the shortest duration of alcohol dependence of all patients (9.9 years). Prevalence of antisocial personality disorder was no different in patients with or without co-occurring ICDs. CONCLUSION: Thirty-eight percent of the alcohol-dependent patients studied presented with an ICD. Patients with ICDs were younger than those without an ICD. The presence of an ICD was not associated with a specific form of alcohol dependence or with antisocial personality. Co-occurrence of pathological gambling, however, was associated with lower age at onset of alcohol dependence, a higher number of detoxifications, and a longer duration of alcohol dependence than was absence of an ICD. PMID- 10362438 TI - Effects of chronic lithium treatment on the peripheral nervous system. AB - BACKGROUND: Although lithium carbonate is widely used in the treatment of mood disorders, symptoms suggesting toxic effects on the peripheral nervous system may emerge even in subjects whose serum lithium levels remain within the recommended therapeutic range. METHOD: Electroneuronographic (ENG) parameters (motor nerve conduction velocity of peroneal and median nerves, sensory nerve conduction velocity of sural and median nerves, amplitude of motor potential of peroneal and median nerves, and amplitude of sensory action potential of the median nerve at the wrist and the sural nerve) were investigated in 2 groups (N = 34) of patients suffering from bipolar affective disorder (DSM-III-R, DSM-IV) undergoing maintenance treatment with lithium carbonate for at least 1 year (mean = 2.06 years) in monotherapy. For 12 patients, ENG results were compared with pretreatment values, whereas in the other 22 cases, only data relevant to posttreatment were available. Fifty-four healthy subjects and 20 patients with recurrent major affective disorder (unipolar and bipolar) never treated with lithium made up the comparison groups. RESULTS: Compared with the 2 comparison groups, patients on chronic lithium treatment showed significant reduction of motor nerve conduction velocity of peroneal and median nerves, sensory nerve conduction velocity of sural and median nerves, amplitude of motor potential of peroneal and median nerves, and amplitude of sensory action potential of the median nerve at the wrist and the sural nerve. The comparison with the assessment made prior to lithium treatment also showed significant changes; after a period of treatment with lithium varying from 2 to 8 years (mean = 5.2 years), significant reductions were found on motor and sensory nerve conduction velocity and on amplitude motor potentials and sensory action potentials. CONCLUSION: Chronic maintenance treatment with lithium affects the peripheral nerves, even if the impairment rarely is such as to warrant discontinuation of treatment. Monitoring of ENG results could be useful for the early detection of neurotoxicity of lithium. PMID- 10362439 TI - Psychotic subtyping of major depressive disorder and posttraumatic stress disorder. AB - BACKGROUND: Many studies have established that a large percentage of patients with posttraumatic stress disorder (PTSD) have comorbid major depressive disorder. Other studies have found that patients with PTSD or a history of childhood trauma have an increased rate of psychotic symptoms. In the present report from the Rhode Island Methods to Improve Diagnosis and Services project, we examine whether an association exists between psychotic subtyping of major depressive disorder and PTSD. METHOD: Five hundred psychiatric outpatients were interviewed with the Structured Clinical Interview for DSM-IV. RESULTS: Almost half of the 500 patients had nonbipolar major depressive disorder (N = 235, 47.0%), 45 (19.1%) of whom had PTSD. Nineteen patients had psychotic depression, 216 had nonpsychotic depression. Compared with patients with nonpsychotic depression, the patients with psychotic depression were nearly 4 times more likely to have PTSD (57.9% vs. 15.7%, Fisher exact test, p = .0001). CONCLUSION: The results of the present study suggest that the presence of psychosis in psychiatric outpatients with major depressive disorder is associated with concurrent PTSD. It is hypothesized that the poorer longitudinal course of psychotic versus nonpsychotic depression may be due to the underrecognition of PTSD in psychotically depressed patients. PMID- 10362440 TI - Risperidone and associated amenorrhea: a report of 5 cases. AB - BACKGROUND: We report a 5-case series in which risperidone use in usual or lower than-usual doses was unexpectedly associated with amenorrhea. CASE REPORTS: On regimens of risperidone (1-8 mg/day), 5 psychiatric patients with various diagnoses developed amenorrhea with elevated serum prolactin levels (mean = 121.7 ng/mL; range, 61.2-229.8 ng/mL). In 4 of 5 cases, menstruation resumed only when risperidone was withdrawn, and in 1 case, menstruation restarted when the dose was tapered. Follow-up serum prolactin levels dropped to a mean of 17.2 ng/mL (range, 6.4-37.6 ng/mL). CONCLUSION: These findings indicate that the occurrence of amenorrhea during risperidone treatment may be related to elevated serum prolactin levels. This phenomenon may be due either to the dopamine D2 blocking effect of risperidone or to the large individual variability in the rate at which risperidone is metabolized. PMID- 10362441 TI - The effects of metrifonate on the cognitive, behavioral, and functional performance of Alzheimer's disease patients. Metrifonate Study Group. AB - BACKGROUND: The objective of this study was to evaluate the efficacy and safety of metrifonate, a long-acting acetylcholinesterase inhibitor, in patients clinically diagnosed with probable Alzheimer's disease of mild-to-moderate severity. METHOD: This was a prospective, multicenter, 26-week, double-blind, parallel group study. The 264 randomized patients met diagnostic criteria of the National Institute of Neurological and Communicative Diseases and Stroke and the Alzheimer's Disease and Related Disorders Association for probable Alzheimer's disease. Patients had Mini-Mental State Examination (MMSE) scores of 10-26 and ischemic scores (Rosen modification) of <4. Metrifonate-treated patients received a single 50-mg dose once daily. The efficacy of metrifonate was investigated with respect to 3 symptom domains. Cognitive performance was analyzed using the Alzheimer's Disease Assessment Scale-Cognitive Subscale (ADAS-Cog) and the MMSE. Psychiatric and behavioral disturbances were analyzed using the Neuropsychiatric Inventory (NPI) and the ADAS-Noncognitive subscale (ADAS-Noncog). The ability to perform instrumental and basic activities of daily living was evaluated using the Disability Assessment for Dementia (DAD) scale. Additionally, global state was assessed using the Clinician Interview-Based Impression of Change with Caregiver Input (CIBIC-Plus) scale. RESULTS: After 26 weeks of metrifonate therapy, a statistically significant benefit of metrifonate was observed in the cognitive performance of Alzheimer's disease patients (ADAS-Cog, t = 2.55, df = 237, p = .012; MMSE, t = 4.60, df = 237, p = .0001). Metrifonate also significantly attenuated the deterioration in activities of daily living of the patients (DAD total score, t = -2.11, df = 233, p = .036) and relieved patients' psychiatric and behavioral disturbances (NPI total score, t = 2.51, df = 233, p = .013). In addition, metrifonate significantly improved the scores for the global state of the patients (CIBIC-Plus, t = 2.07, df = 232, p = .039). Metrifonate was well tolerated; adverse events were predominantly mild in intensity, and no hepatotoxicity was observed. CONCLUSION: In this study, metrifonate was safe and well tolerated. It benefited the cognitive decline, psychiatric and behavioral disturbances, impaired ability to perform instrumental and basic activities of daily living, and global state of patients diagnosed with mild-to-moderate Alzheimer's disease. PMID- 10362442 TI - Efficacy of SSRIs and newer antidepressants in severe depression: comparison with TCAs. AB - BACKGROUND: The significant morbidity and mortality associated with severe depression and its psychotic or melancholic subtypes necessitate effective and well-tolerated therapy. This review evaluates antidepressant treatments for patients with severe depression. DATA SOURCES: Comparative clinical trials conducted on patients with severe depression were found by an English-language MEDLINE search (1985 to present). Additional studies were identified in article bibliographies. Search terms included depressive disorders, depression and severe, hospitalized, melancholic or melancholia, psychotic, and endogenous. STUDY FINDINGS: Evidence for efficacy of SSRIs in severe or melancholic depression comes from a small but growing number of controlled studies with adequate samples, as well as meta-analyses and retrospective subgroup analysis of premarketing trials. In studies that defined response as a 50% or greater reduction in Hamilton Rating Scale for Depression (HAM-D) scores, response rates ranged from 53% to 64% for SSRIs and 43% to 70% for TCAs. In separate trials on severe depression, venlafaxine and mirtazapine were both more effective than placebo and an active comparator. Nefazodone and bupropion were each found to be more effective than placebo in studies of severe depression. Venlafaxine and mirtazapine have been found to be more effective than fluoxetine. CONCLUSION: SSRIs and TCAs are comparably effective for the treatment of severe or melancholic depression. SSRIs and other newer agents appear to be better tolerated than TCAs, specifically lacking adverse anticholinergic and cardiovascular effects that may limit the use of TCAs. Emerging data with venlafaxine and mirtazapine in severely depressed patients with or without melancholia support the efficacy of these treatments. Nefazodone and bupropion were found to be effective in hospitalized depressed patients. Electroconvulsive therapy (ECT) or combined antidepressant therapy may be useful in some patients with severe depression. Patients with severe psychotic depression may respond better to an antipsychotic-antidepressant combination. PMID- 10362443 TI - Weight gain and antipsychotic medications. PMID- 10362444 TI - Ileus as a possible result of bupropion in an elderly woman. PMID- 10362445 TI - Methamphetamine-associated obsessional symptoms and effective risperidone treatment: a case report. PMID- 10362446 TI - Comparing the efficacy and safety of fluoxetine and venlafaxine in outpatient depression. PMID- 10362447 TI - Relationship between antidepressant treatment and suicide. PMID- 10362448 TI - Risperidone treatment of tardive dyskinesia and dystonia. PMID- 10362449 TI - Patterns of remission and relapse in obsessive-compulsive disorder: a 2-year prospective study. AB - OBJECTIVE: This study examined the course of illness in patients with obsessive compulsive disorder (OCD) over a 2-year period. METHOD: Sixty-six patients with a primary diagnosis of DSM-III-R OCD were followed prospectively for 2 years. Baseline information was collected on demographic characteristics, Axis I and II diagnoses, and severity of OCD symptoms. Follow-up measures obtained at 3, 6, 12, and 24 months after baseline assessment included information on symptomatic and diagnostic status as well as behavioral and somatic treatments received. RESULTS: The probability of full remission from OCD over the 2-year period was 12%. The probability of partial remission was 47%. After achieving remission from OCD, the probability of relapse was 48%. No factors were identified that significantly predicted full or partial remission. Seventy-seven percent (N = 51) of the subjects received a serotonin reuptake inhibitor (SRI) for > or =12 weeks, and 68% (N = 45) received medium-to-high doses of SRIs for > or =12 weeks. Only 18% received a full trial of behavior therapy. CONCLUSION: Despite exposure to at least 1 adequate trial of an SRI, the likelihood of full remission of OCD in this study was low. Results of this study also suggest that behavior therapy may be under-utilized. PMID- 10362450 TI - The effects of zinc depletion on peak force and total work of knee and shoulder extensor and flexor muscles. AB - In this study we tested the effect of zinc (Zn) on muscle function in human. After receiving 12 mg Zn/day for 17 days, 8 male subjects received 0.3 mg Zn/day for either 33 or 41 days. Subjects were divided into two groups for repletion. Group A subjects received overnight infusions of 66 mg Zn on Days 1 and 10 and then were fed 12 mg Zn/day for another 16 days. Group B subjects were fed 12 mg Zn/day for 3 weeks. Peak force and total work capacity of the knee and shoulder extensor and flexor muscle groups were assessed using an isokinetic dynamometer at baseline, at two points during depletion, and at repletion. Plasma Zn declined significantly during depletion and remained below baseline levels after repletion. The peak force of the muscle groups tested was not affected by acute Zn depletion, however, total work capacity for the knee extensor muscles and shoulder extensor and flexor muscles declined significantly. The data suggest that acute Zn depletion alters the total work capacity of skeletal muscle. PMID- 10362451 TI - Creatine supplementation differentially affects maximal isometric strength and time to fatigue in large and small muscle groups. AB - Ten physically active, untrained, college-aged males (26.4 +/- 5. 8 years old) received creatine (CR, 5 g creatine monohydrate + 3 g dextrose) and placebo (PLA, 7 g dextrose) supplementation four times per day for 5 days in a double-blind, randomized, balanced, crossover design. Performance was assessed during maximal and three repeated submaximal bouts of isometric knee extension and handgrip exercise. CR supplementation significantly increased (p <.05) maximal isometric strength during knee extension but not during handgrip exercise. CR supplementation increased time to fatigue during each of the three bouts of submaximal knee extension and handgrip exercise when compared to the PLA trials. These findings suggest that CR supplementation can increase maximal strength and time to fatigue during isometric exercise. However, the improvements in maximal isometric strength following CR supplementation appear to be restricted to movements performed with a large muscle mass. PMID- 10362452 TI - Effects of in-season (5 weeks) creatine and pyruvate supplementation on anaerobic performance and body composition in American football players. AB - The purpose of this investigation was to study the efficacy of two dietary supplements on measures of body mass, body composition, and performance in 42 American football players. Group CM (n = 9) received creatine monohydrate, Group P (n = 11) received calcium pyruvate, Group COM (n = 11) received a combination of calcium pyruvate (60%) and creatine (40%), and Group PL received a placebo. Tests were performed before (T1) and after (T2) the 50 week supplementation period, during which the subjects continued their normal training schedules. Compared to P and PL, CM and COM showed significantly greater increases for body mass, lean body mass, 1 repetition maximum (RM) bench press, combined 1 RM squat and bench press, and static vertical jump (SVJ) power output. Peak rate of force development for SVJ was significantly greater for CM compared to P and PL. Creatine and the combination supplement enhanced training adaptations associated with body mass/composition, maximum strength, and SVJ; however, pyruvate supplementation alone was ineffective. PMID- 10362453 TI - Effect of Q10 supplementation on tissue Q10 levels and adenine nucleotide catabolism during high-intensity exercise. AB - The aim of the present study was to investigate the concentration of ubiquinone 10 (Q10), at rest, in human skeletal muscle and blood plasma before and after a period of high-intensity training with or without Q10 supplementation. Another aim was to explore whether adenine nucleotide catabolism, lipid peroxidation, and mitochondrial function were affected by Q10 treatment. Seventeen young healthy men were assigned to either a control (placebo) or Q10-supplementation (120 mg/day) group. Q10 supplementation resulted in a significantly higher plasma Q10/total cholesterol level on Days 11 and 20 compared with Day 1. There was no significant change in the concentration of Q10 in skeletal muscle or in isolated skeletal muscle mitochondria in either group. Plasma hypoxanthine and uric acid concentrations increased markedly after each exercise test session in both groups. After the training period, the postexercise increase in plasma hypoxanthine was markedly reduced in both groups, but the response was partially reversed after the recovery period. It was concluded that Q10 supplementation increases the concentration of Q10 in plasma but not in skeletal muscle. PMID- 10362454 TI - Metabolic effects of a protein-supplemented carbohydrate drink in marathon runners. AB - A field study was performed to investigate the acute influence of a milk protein hydrolysate supplemented drink (CHO+PRO) on metabolism during and after a marathon run compared to the same drink without protein (CHO). Carbohydrate metabolites and hormones were not influenced by CHO+PRO. Levels of plasma free fatty acids were significantly lower and levels of urea and most amino acids were significantly higher with CHO+PRO. Sweat urea and ammonia nitrogen excretion during the run as well as urinary 3-methylhistidine excretion during the entire exercise day was similar in both treatments. Urinary total nitrogen was significantly increased and urinary pH decreased with CHO+PRO. It was concluded that the supplemented protein was absorbed and probably at least partially oxidized during the run and that no obvious negative metabolic effects occurred. CHO+PRO did not acutely affect myofibrillar protein breakdown as assessed by the 3-methylhistidine method; however, total body protein breakdown was not measured. PMID- 10362455 TI - Effective nutritional ergogenic aids. AB - Athletes use a variety of nutritional ergogenic aids to enhance performance. Most nutritional aids can be categorized as a potential energy source, an anabolic enhancer, a cellular component, or a recovery aid. Studies have consistently shown that carbohydrates consumed immediately before or after exercise enhance performance by increasing glycogen stores and delaying fatigue. Protein and amino acid supplementation may serve an anabolic role by optimizing body composition crucial in strength-related sports. Dietary antioxidants, such as vitamins C and E and carotenes, may prevent oxidative stress that occurs with intense exercise. Performance during high-intensity exercise, such as sprinting, may be improved with short-term creatine loading, and high effort exercise lasting 1-7 min may be improved through bicarbonate loading immediately prior to activity. Caffeine dosing before exercise delays fatigue and may enhance performance of high intensity exercise. PMID- 10362456 TI - Peroxisome proliferator activated receptor-gamma, leptin and tumor necrosis factor-alpha mRNA expression during very low calorie diet in subcutaneous adipose tissue in obese women. AB - BACKGROUND: PPAR gamma, leptin and TNF alpha are three major factors that play a key role in influencing adipocyte differentiation and both adipose tissue function and metabolism. However, the regulation of these three genes during a dynamic period of weight loss is unknown. We therefore investigated the concomitant regulation of the mRNA expression of PPAR gamma, leptin and TNF alpha in adipose tissue during a 21-day very low calorie diet (VLCD) in 12 non-diabetic obese women. METHODS: The mRNA levels of PPAR gamma, leptin and TNF alpha were quantified by quantitative RT-competitive PCR in abdominal subcutaneous adipose tissue before and during VLCD (940 kcal/day). RESULTS: VLCD induced weight loss (approximately 6 kg) and improved insulin sensitivity. Simultaneously, VLCD induced the reduction in the adipose tissue mRNA abundances of PPAR gamma (-13%, p < 0.05) and of leptin (-58%, p < 0.005), whereas TNF alpha mRNA levels increased (+78%, p < 0.005). PPAR gamma and leptin mRNA levels were correlated before (r = 0.778, p < 0.01) and after VLCD (r = 0.797, p < 0.01). Serum HDL cholesterol concentrations were positively associated with PPAR gamma (r = 0.696, p < 0.03) and leptin (r = 0.806, p < 0.01) mRNA levels. CONCLUSIONS: The increase in TNF alpha mRNA levels suggested that a local increased expression of this cytokine in adipose tissue might play a role in the control of the fat mass during weight loss. PPAR gamma and leptin mRNA levels were positively associated both before and after VLCD, suggesting that common regulatory mechanism(s) might control their expression. More strikingly, we found strong positive correlations between circulating HDL-cholesterol and both PPAR gamma and leptin mRNA levels, suggesting the existence of physiological links between circulating lipoprotein metabolism and adipose tissue function. PMID- 10362457 TI - Arterial hypertension and glycemia in non-diabetic subjects: is there an association independent of obesity? AB - BACKGROUND: A possible association of glycemia with arterial hypertension has been suggested by the frequent co-occurrence of impaired glucose tolerance or Type 2 diabetes mellitus with arterial hypertension. The objective was to examine the relationship of glycated hemoglobin (HbA1c) concentration with arterial hypertension status in non-diabetic subjects. METHODS: A cross-sectional analysis of baseline data from the EPIC-Potsdam Cohort Study, Germany, was performed. The study population comprised 1846 non-diabetic subjects, 772 men and 1074 women, age 35-65. Blood pressure was measured three times consecutively. Level of HbA1c was determined by an assay based on monoclonal antibodies. Body height, weight and circumferences were obtained. Arterial hypertension status was either determined through blood pressure measurement (blood pressure > or = 160/95 mmHg) or based on antihypertensive drug use. HbA1c was divided into sex-specific quintiles and logistic regression was used to estimate the odds of being hypertensive and the corresponding confidence intervals. RESULTS: The highest compared to the lowest quintiles of HbA1c were in univariate analysis associated with being hypertensive. Adjustment for age and body mass index completely removed any significant association with arterial hypertension status. The odds ratio in men was 1.1 (95% CI 0.7-1.8), and in women it was 0.9 (95% CI 0.5-1.4). Repeating the analysis with systolic and diastolic blood pressure among untreated hypertensives yielded similar results. CONCLUSION: Unlike previous studies, our data do not support an association of HbA1c with arterial hypertension that is statistically independent of age and body mass index. Whether these established arterial hypertension risk factors are truly confounders of the HbA1c-arterial hypertension association or rather potentially antecedent factors requires further study. PMID- 10362458 TI - Changes induced by sucrose administration upon the morphology and function of pancreatic islets in the normal hamster. AB - BACKGROUND: This report documents sequential changes in islet morphology (cell replication and islet neogenesis) and glucose-induced insulin secretion in young normal male Syrian hamsters. METHODS: Three-week-old animals received a control standard commercial diet or this diet supplemented with sucrose--10% (w/v) solution in drinking water, a treatment that stimulated pancreatic growth and function--for 5 (C5/S5) or 21 (C21/S21) weeks. Insulin secretion and content were measured in isolated islets, while several biochemical parameters were assessed in serum. Different morphological features were analysed in the endocrine pancreas by quantitative immunocytochemistry. RESULTS: Serum glucose, triglycerides and total cholesterol levels were comparable among the groups, whereas serum- and pancreatic-insulin levels were higher in the S hamsters. Islets from S21 hamsters released more insulin than those from C21 animals at all glucose concentrations tested. The volume densities of the total endocrine pancreas (1.9 +/- 0.2 vs 1.2 +/- 0.2; p < 0.02) of the beta-cell subpopulation, the islet number per unit area (2.4 +/- 0.1 vs 1.2 +/- 0.1; p < 0.0004) and the beta-cell mass (4.2 +/- 0.5 vs 2.3 +/- 0.5; p < 0.01) were significantly higher in S5 vs C5 animals. Conversely, the islet volume and the number of beta cells/islets were significantly smaller in S5 than in C5 animals. The beta-cell replication rate in S5 hamsters was 10-fold that of C5 animals. All these parameters had comparable values in S21 and C21 animals. We detected cytokeratin labelled cells located at the islet periphery (in alpha cells) and among the ductular cells, only in the S5 hamsters. CONCLUSIONS: Sucrose administration to young hamsters causes time-dependent pancreatic modifications, with morphological changes (increase in islet- and in beta-cell mass with incremented beta-cell replication rate and evidence of islet neogenesis) occurring at 5 weeks and insulin secretion (increase in insulin sensitivity to glucose) being mainly affected at 21 weeks. This experimental model could prove useful for studying the mechanisms underlying the control of islet-cell population distribution and for developing new strategies in preventing cell damage. PMID- 10362460 TI - New insights into the epidemiology of type 1 diabetes in Mediterranean countries. AB - In Mediterranean countries, the incidence (per 100,000 per year) of Type 1 diabetes in children aged under 15 years shows wide variation from country to country, ranging from 2.45 in Macedonia to 34.4 in Sardinia. By interacting with environmental factors such as diet, toxins or viral infections, the HLA plus non HLA genes of susceptibility or resistance to Type 1 diabetes so far identified are the strongest determinants of the disease as far as incidence, age at onset and sex ratio are concerned. The distribution of these genes in the Mediterranean region is still not completely known. Sardinians are the most suitable population for studying such phenomena for three main reasons: their genetic features have been repeatedly analysed in depth; their incidence rate of Type 1 diabetes is by far the highest in the Mediterranean area; and considerable colonies of Sardinian emigrants settled in neighbouring low-incidence Italian regions. Studies on Spaniards and Jews have also contributed to a better understanding of the respective roles of genetic and environmental factors. From a body of research on the Mediterranean populations, it can be concluded that great genetic heterogeneity accounts for the wide variation in incidence of Type 1 diabetes, while rather ubiquitious environmental factors trigger the disease in genetically predisposed individuals. PMID- 10362459 TI - Migrant populations and the incidence of type 1 diabetes mellitus: an overview of the literature with a focus on the Spanish-heritage countries in Latin America. AB - Type 1 diabetes mellitus (DM) is a 'chronic' autoimmune disorder leading to the destruction of the pancreatic beta cell. The natural history of diabetes includes a long subclinical (prediabetes) period. The pathogenesis is multifactorial and characterized by the interaction of environmental factors, with predisposing genes, most of which are associated with the HLA DR DQ loci. The relatively recent development of worldwide incidence registries for Type 1 DM has allowed us to compare the epidemiological results obtained in most parts of the world. This approach is particularly valuable in analysing the effects of migration of populations from one area of the world where the incidence of Type 1 DM is different (usually lower) to a new geographic setting. Properly designed migrant studies may be valuable in uncovering whether the genetic background remains more important than the new 'exposure' as illustrated by the Sardinian migration to Lazio and Lombardy. The presence of some putative 'protective' environmental exposures or the absence of those prevalent in the country of origin may explain the usually lower Type 1 DM incidence observed in most countries (Chile, Peru, Mexico) sharing a 'Spanish caucasoid genetic pool', and even in relatively genetically homogeneous groups such as Japanese populations migrating to Hawaii. In fact, the disease is caused by both genetic and environmental factors and to convince the scientific community of this fact is a primary responsibility for epidemiologists. PMID- 10362461 TI - Immune markers for monitoring the progression of autoimmune disease. AB - The incidence of immune-mediated diseases is increasing worldwide. Reliable immune markers for monitoring disease progression and also the effect of therapeutic interventions are urgently needed in order to investigate preventive or therapeutic measures effectively. At a recent workshop held on 5 December 1998 in Copenhagen, the state of research on surrogate markers in Type 1 diabetes was discussed and compared with the experience in multiple sclerosis, inflammatory bowel disease and transplantation. PMID- 10362462 TI - The management of hypertension in a diabetic pregnancy. AB - Pregnancy in a woman with Type 1 diabetes poses several clinical challenges. In addition to meticulous glycaemic control, careful attention must be paid to the management of developing and pre-existing diabetic complications which may progress in severity during pregnancy. Pregnancy-induced hypertension is more common in women with diabetes and especially in those with diabetes of long duration. Diabetic renal disease is associated with hypertension which often deteriorates during pregnancy. The management of hypertension is difficult because of limited therapeutic options and the need to consider the implications for the developing fetus as well as the mother. This case report details the clinical management of a young woman with Type 1 diabetes whose pregnancy was complicated by the development of hypertension. PMID- 10362463 TI - Diabetes: a strategic plan for the 21st century. PMID- 10362464 TI - Ultrasound-assisted analysis of total carbohydrates in environmental and food samples. AB - Analytical methods to determine total carbohydrates in environmental and food samples usually require a preliminary chemical hydrolytic procedure to convert polysaccharides into monosaccharides prior to detection by colorimetric or chromatographic techniques. In this paper, an alternative hydrolytic procedure is presented. The method for hydrolysis of polysaccharides is based on the application of ultrasound at room temperature. The advantages of the method proposed here are reduced time required for analysis (4-5 h) and the improvement of the analytical accuracy due to the absence of the most common reactions of oxidation. The proposed method was applied to the determination of total carbohydrates in several environmental samples (seawater and marine mucilage) and starchy food samples. Results for total carbohydrate obtained using the ultrasound procedure for hydrolysis of samples agreed with those found when applying other published procedures. PMID- 10362465 TI - Numerical Solution of the Extended Nernst-Planck Model. AB - The main features of a numerical model aiming at predicting the drift of ions in an electrolytic solution upon a chemical potential gradient are presented. The mechanisms of ionic diffusion are described by solving the extended Nernst-Planck system of equations. The electrical coupling between the various ionic fluxes is accounted for by the Poisson equation. Furthermore, chemical activity effects are considered in the model. The whole system of nonlinear equations is solved using the finite-element method. Results yielded by the model for simple test cases are compared to those obtained using an analytical solution. Applications of the model to more complex problems are also presented and discussed. Copyright 1999 Academic Press. PMID- 10362466 TI - The Micelle-Induced Interaction between Ninhydrin and Tryptophan. AB - The effects of micelles of cetyltrimethylammonium bromide (CTAB) and cetylpyridinium bromide (CPB) on the observed pseudo-first-order rate constants for the interaction of ninhydrin with tryptophan were studied. The influence of different parameters was considered, i.e., reactant concentration, surfactant concentration, temperature, and effect of added salts. The data are explained in terms of the pseudo-phase model of the micelles. Copyright 1999 Academic Press. PMID- 10362467 TI - Micellar Binding of alpha-Diimine-Chromium(III) Complexes (Cr(NN)3+3) to Sodium Dodecyl Sulfate (SDS). AB - Binding interactions of alpha-diimine-chromium(III) complexes with sodium dodecyl sulfate (SDS) were studied in air-saturated solution and in N2-purged solution using lifetime measurements. The lifetime titration curves consisted of two regions with different slopes: one of them negative and more pronounced at low concentrations of SDS until reaching a minimum and the other, at a later stage, with a less pronounced positive slope till a plateau was reached. This biphasic behavior of lifetime-SDS concentration data revealed the presence of premicellar aggregates at low SDS concentration and the formation of normal micelles at high surfactant concentration. The results were analyzed with a model that includes binding of the sensitizer to micelles and to small premicellar aggregates. Binding constants obtained by fitting of lifetime titration curves are in agreement with the electrostatic interactions between the complexes and the surfactant and with the hydrophobic interactions between the ligands of the complexes and the formed micelle. Copyright 1999 Academic Press. PMID- 10362468 TI - The Structure of Amorphous Bulk and Silica-Supported Copper(II) Hydroxides. AB - The data obtained show that at pH 7 copper(II) ions are adsorbed on a SiO2 surface as polymeric species of hydroxide nature. The structure of these species is similar to that of the bulk amorphous copper hydroxide. The amorphous state of supported Cu(OH)2 is caused by a small (ca. 11 A) size of the surface particles. In contrast, the overstoicheometric water molecules seem to act as "amorphizers" of the bulk copper hydroxide. The structures of the bulk and dispersed amorphous copper(II) hydroxide were determined. The amorphous Cu(OH)2 has a layered structure close to the structure of the crystalline hydroxide, but the layers in the amorphous hydroxide are shifted toward one another approximately for 14 of the "c" period of the lattice. Copyright 1999 Academic Press. PMID- 10362469 TI - Marginal Regeneration and the Marangoni Effect. AB - On the basis of experimental observations described earlier, we have proposed that marginal regeneration is caused by surface tension gradients at the borders of mobile foam films. Marginal regeneration is the rate-determining mechanism in the drainage of such films, and, as such, a determining factor in the persistence (or long-term stability) of foams. Marangoni flows are caused by surface tension gradients, and these set off the exchange of thicker for thin film elements along the borders, while the total film area remains the same. In this paper we present simulations of the drainage of liquid in a vertical soap film, and show that it is realistic to expect large surface tension gradients along the lower border of the film under the conditions which lead to marginal regeneration. Copyright 1999 Academic Press. PMID- 10362470 TI - Adsorption of Telechelic Poly(ethylene oxide) on Colloids: Influence on Colloid Stability. AB - Adsorption of telechelic polymers (hydrophobically end-capped poly(ethylene oxide)) on charged colloidal particles modifies the characteristics of the electrical double layer. These interfaces were found to be able to protect colloids against aggregation or to induce colloid destabilization depending on the molecular weight and type of surface coating. Colloid stability is observed when the polymer layer is thinner than the electrical double layer and when the adsorbed polymer adopts a tethered brush structure. Aggregation of coated latex particles induced by formation of interparticle polymer links is observed when (i) the distance of the random walk of the dangling paraffinic end group is larger than the thickness of the electrical double layer and (ii) the adsorbed polymer brush adopts a loopy structure. Aggregation is attributed to adsorption of hydrophobic end chains on two different latex particles. Copyright 1999 Academic Press. PMID- 10362471 TI - Influence of the Fractal Character of Model Substances on their Reactivity at Solid-Liquid Interfaces. AB - The fractal theory has been applied to study surface complexity where surface aspects influence physical properties and play a role in controlling heterogeneous reactions at interfaces. In this work, the influence of the fractal character of some selected antacids was investigated in regard to their neutralizing activity. The materials used were magnesium trisilicate, magnesium hydroxide, and heavy and light magnesium oxide, each from three different manufacturers. Surface area, total pore volume, and particle size were measured. Fractal dimension was determined from gas adsorption data according to pore size distribution, the Frenkel-Halsey-Hill, and thermodynamic methods. The results obtained show a correlation between neutralization activity and fractal character rather than total surface area or particle size. Also, the effect of porosity in terms of total pore volume was modified by the structure of the porous network. The complexity of the pore network played a major role in controlling reactivity. However, the effect of surface roughness was only demonstrated when adsorption on the surface was a rate-limiting step in the reaction. Copyright 1999 Academic Press. PMID- 10362472 TI - Effect of Pore Clogging on Kinetics of Lead Uptake by Clinoptilolite. AB - The kinetics of lead-sodium ion exchange using pretreated natural clinoptilolite are investigated, more specifically the influence of agitation (0, 210, and 650 rpm) on the limiting step of the overall process, for particle sizes of 0.63-0.8 and 0.8-1 mm at ambient temperature and initial lead solutions of 500 mg l-1 without pH adjustment. The isotopic exchange model is found to fit the ion exchange process. Particle diffusion is shown to be the controlling step for both particle sizes under agitation, while in the absence of agitation film diffusion is shown to control. The ion exchange process effective diffusion coefficients are calculated and found to depend strongly on particle size in the case of agitation at 210 rpm and only slightly on particle size at 650 rpm. Lead uptake rates are higher for smaller particles only at rigorous agitation, while at mild agitation the results are reversed. These facts are due to partial clogging of the pores of the mineral during the grinding process. This is verified through comparison of lead uptake rates for two samples of the same particle size, one of which is rigorously washed for a certain time before being exposed to the ion exchange. Copyright 1999 Academic Press. PMID- 10362473 TI - Micelle Formation of Anionic Surfactant with Divalent Counterion of Separate Electric Charge. AB - The critical micelle concentration (CMC) of the anionic surfactant, 1,1'-(1,omega decanediyl) bispyridinium hexadecane-1-sulfonate (C10BP(C16)2) was determined by electrical conductivity measurements at various temperatures. The degree of counterion binding to micelles was evaluated from the change in CMC with total counterion concentration. The molecular weight of the micelles was determined by static light scattering. The mass action model was applied to micelle formation in order to calculate the three micellization parameters, the micellization constant, the micelle aggregation number, and the number of counterions per micelle. Thermodynamic parameters (DeltaG0, DeltaH0, -TDeltaS0) for the micellization were evaluated by their temperature dependence. The findings were: (1) Micelle formation was entropy-driven over the whole temperature range examined. (2) C10BP(C16)2 had a higher degree of counterion binding to micelles compared with those of monovalent counterion. (3) The plots of log CMC against the carbon number of the homologous surfactant ions gave a straight line, indicating that free energy change per methylene group for micelle formation was 1.18RT for surfactant ions. Copyright 1999 Academic Press. PMID- 10362474 TI - Reactions of p-Nitrophenyl Diphenyl Phosphinate with Fluoride and Hydroxide Ion in Nonionic Micelles: Kinetic Salt Effects. AB - Reactions of p-nitrophenyl diphenyl phosphinate (p-NPDP) with OH- and F- are inhibited by micelles of dodecyl polyoxyethylene ethers C12E10 and C12E23. Rate constants decrease sharply in very dilute surfactant and become approximately constant as p-NPDP becomes fully micellar bound, with inhibition by approximately an order of magnitude. The first-formed fluoridate is detected by NMR spectroscopy in reaction with F- in DMSO-d6:D2O 85:15 v/v, but not in other conditions. Kinetic salt effects on the micellar reactions are ion specific. Bulky anions, ClO-4 and naphthalene-2-sulfonate ion (ONs-), inhibit reactions much more than small ions, and (C7H15)4N+ accelerates reaction of F-. Micellar incorporation of ONs- in the micelles is demonstrated by changes in 1H-NMR spectra of the surfactants. Comparison of rate constants of reaction of F- in PEG (MW 10(4)) and in nonionic micelles shows that the micellar inhibition is due largely to lower ionic concentrations in the micelles than in water. Copyright 1999 Academic Press. PMID- 10362475 TI - Size Distributions out of Static Light Scattering: Inclusion of Distortions from the Experimental Setup, e.g., a SOFICA-type Goniometer. AB - In this paper the estimation of sphere size distributions of polymer latex with static light scattering is investigated. For the calculation of the scattering data a model is proposed which describes a SOFICA-type goniometer that was used for the light scattering experiments. From the comparison of this model with a model that is based on Mie's Theory only, conclusions about the reliability of the estimation of size distributions from uni- and bimodal colloidal suspensions with static light scattering could be drawn. Furthermore, the contribution of multiple scattering was investigated, and a method is suggested which further improves the obtained results. Both simulated and experimental data were examined. For data analysis, a method based on Tikhonov regularization was used. Copyright 1999 Academic Press. PMID- 10362476 TI - A Lagrangian Stochastic Model for Heavy Particle Deposition. AB - A Lagrangian stochastic model for the deposition of heavy particles from turbulent flows is presented. Heavy particles are treated as tracer particles moving in a virtual fluid having heavy particle velocity statistics. These velocity statistics are deduced from the particle momentum equation. The model satisfies the well-mixed condition for this virtual fluid and is consistent with similarity theory. Model agreement with experimental data for the "inertia moderated" regime and the upper half of the "diffusion-impaction" regime (nondimensional particle relaxation time, tau+p > 5) is excellent. In accord with the results of direct numerical simulations for the diffusion-impaction regime, the model predicts a build-up of particle concentration in the viscous sublayer. Such a build-up concentration is not predicted by current Lagrangian stochastic models nor by models utilizing the "diffusion/free-flight" concept. Copyright 1999 Academic Press. PMID- 10362477 TI - Surface Chemistry Studies of Photosystem II. AB - The monolayer systems of PS II membranes, PS II core complex particles, and the mixture of monogalactosyldiacylglycerol (MGDG) with PS II core complex particles were constructed by means of the Langmuir technique. The surface pressure and surface potential properties of these systems were analyzed. The apparent particle sizes were determined from surface pressure-area isotherms, which were 200 nm2 for PS II membranes and 320 nm2 for PS II core complex particles. The surface pressure isotherm analysis of the mixed monolayer of MGDG and PSII core complex particles shows that the mixed monolayer has good miscibility at all surface pressures. Copyright 1999 Academic Press. PMID- 10362478 TI - Rheological Behavior of Suspensions Flocculated by Weak Bridging of Polymer Coils. AB - A polymer coil with weak affinity for the solid surface makes a flexible bridge between particles by reversible adsorption. When the coil size is comparable to particle diameter in solution, the suspensions flocculated by reversible bridging show shear-thickening flow in a narrow range of shear rates. The force generated by the rapid extension of bridges within the lifetime is responsible for the shear thickening. Although it is expected that the bridges become more flexible with increasing coil size, the polymers with high molecular weights do not necessarily give rise to shear thickening. The striking shear thickening is induced by polymers with molecular weights, Mw, of 2.5 x 10(5) and 4.5 x 10(5). The viscosity enhancement in the shear-thickening region is considerably suppressed for polymers with Mw = 7.5 x 10(5). From the sedimentation experiments, a large portion of the segments of a coil with Mw = 7.5 x 10(5) is adsorbed in train on the particle surface. The addition of small amounts of surfactant leads to an increase in the fraction of loops at the expense of trains and the bridges become more flexible. As a result, the flow of suspensions in the presence of surfactant becomes shear thickening. The rheology of suspensions flocculated by reversible bridging can be modified by a balance between loop and train fractions in a bridge. Copyright 1999 Academic Press. PMID- 10362479 TI - Structure Formation in Polymer-Modified Liquid Crystals. AB - The objective is the investigation of the influence of two different polymers in a lamellar liquid crystalline sodium dodecyl sulfate (SDS)/decanol/water system. The polymers are, on the one hand, uncharged poly(ethylene glycol) (PEG), whose molecular weight is varied, and, on the other hand, a polycation with differing charge densities made of diallyldimethylammonium chloride (DADMAC) and N-methyl-N vinylacetamide (NMVA). Applied methods were mainly small-angle X-ray diffraction (SAXS), rheology, and electron microscopy. Our major results are the observation of two coexisting lamellar structures of which one is a "nonswelling" and the other a "swelling" structure, which form multilamellar spherical structures on a supramolecular level. However, the variation of the molecular weight of PEG can be neglected, whereas the alteration of the charge density has a pronounced influence on the structure. Generally, one can conclude that the phenomena observed here can be understood on the basis of polymer-polymer interactions in contrast to polymer-surfactant interactions of preliminary electrostatic nature. Copyright 1999 Academic Press. PMID- 10362480 TI - Small-Angle Neutron Scattering and Fluorescence Studies of Mixed Surfactants with Dodecyl Tails. AB - The mixed micelles formed in solutions of the anionic surfactant sodium dodecylsulfate (SDS) and the nonionic surfactant dodecylmalono-bis-N methylglucamide (DBNMG) were studied by surface tension, fluorescence, and small angle neutron scattering. Measurements of the critical micelle concentration (CMC) by surface tension and fluorescence show that these two surfactants mix nonideally. The experimental values of the CMCs for surfactant mixtures for all compositions are less than the ideal prediction proposed by Clint (J. Chem. Soc. 71, 1327 (1975)). Regular solution theory (for example, Penfold et al., Langmuir 11, 2498 (1995)) can be used to calculate the composition of the micelle at the CMC. The micellar composition thus calculated differs significantly from that at higher total surfactant concentrations, the latter being much closer to the stoichiometric composition. The polarity of the micellar environment, as perceived by a solubilized fluorescence probe, is identical for both the single component and the mixed micelles. The scattering data show that the mixed micelles are comparable in size to the pure nonionic micelle and slightly larger than the anionic micelle. Parallel studies with deuterated SDS showed no evidence of segregation or local ordering within the mixed micelle. The effective charge on the mixed micelle is determined by the amount of ionic surfactant within the micelle. Addition of 0.1 M NaCl to these systems has no effect on the CMC, micelle ellipticity, and aggregation number-a behavior very different from that observed for simple SDS solutions. Copyright 1999 Academic Press. PMID- 10362481 TI - The Influence of Interfacial Structuring on Gibbsite Interactions in Synthetic Bayer Liquors. AB - The interaction forces between gibbsite particles aging in synthetic, supersaturated sodium and potassium aluminate solutions (Bayer liquors) have been investigated. Compliance with DLVO-type behavior was observed in dilute electrolyte. Under synthetic Bayer conditions, non-DLVO, repulsive steric forces exist. Upon aging, this steric repulsion gradually diminishes and disappears. A structured interfacial layer, displaying inner and outer regions of different compressibility, develops as aging occurs and interparticle adhesion increases. At high particle approach velocities, hydrodynamic forces exist which are dependent upon the viscosity of solution. The steric, adhesive, and hydrodynamic interactions are all greater in NaAl(OH)4 compared with KAl(OH)4 liquors of the same concentrations and degree of aging. Solution Al(OH)-4 ion-mediated interfacial structuring is apparently responsible for the time-dependent repulsive and adhesive forces. These interfacial phenomena are perceived to be precursors to, or part of, gibbsite crystal growth and agglomeration mechanisms. Copyright 1999 Academic Press. PMID- 10362482 TI - Comparison of the Poisson-Boltzmann Model and the Donnan Equilibrium of a Polyelectrolyte in Salt Solution. AB - A sequence of charged parallel flat plates immersed in a salt solution is used to model the swelling behavior of a polyelectrolyte. The solution of the nonlinear Poisson-Boltzmann equation gives rise to an analytical expression of the plate distance as a function of swelling pressure (p), plate charge density (varsigma), and salt concentration (n). The exact treatment of the system also yields the connection between polyelectrolyte concentration nP, p, varsigma, and n. A formula for the electric potential difference between polyelectrolyte and salt solutions is derived. On the other hand, the relation between nP, p, and n is known from the Donnan equilibrium. It is shown that the Donnan potential, UD(nP, n), and swelling pressure, pD(nP, n), are always larger than the equivalent quantities in the Poisson-Boltzmann theory (UPB(varsigma, nP, n) and pPB(varsigma, nP, n), respectively). The transition from the Poisson-Boltzmann theory to the Donnan model is achieved by the limiting process varsigma --> 0 which reveals the intrinsic linkage between the two theories. pD(nP, n) = limvarsigma-->0 pPB(varsigma, nP, n) and UD(nP, n) = limvarsigma-->0 UPB(varsigma, nP, n). Copyright 1999 Academic Press. PMID- 10362483 TI - Sorption of Metal Ions on Clay Minerals. AB - The local structural environment of Co sorbed on hectorite (a magnesian smectite) has been investigated by polarized EXAFS (P-EXAFS) spectroscopy on a self supporting film of Co-sorbed hectorite. This sorption sample was prepared by contacting Co and hectorite at pH 6.5 and at high ionic strength (0.3 M NaNO3) to favor pH-dependent sorption reaction over cation exchange. A self-supporting film was elaborated after 120 h of reacting time, when apparent quasi-equilibrium conditions were attained. The half-width at half maximum of the orientation distribution of c* axis of individual clay platelets off the film normal was determined by quantitative texture analysis, and found to be equal to 18.9 degrees. Co K-edge P-EXAFS spectra were recorded at angles between the incident beam and the film normal equal to 0 degrees, 35 degrees, 50 degrees, and 60 degrees; the 90 degrees spectrum was obtained by extrapolation. Spectral analysis led to the identification of the two nearest cationic subshells containing 1.6 +/ 0.4 Mg at 3.03 A and 2.2 +/- 0.5 Si at 3.27 A. These distances are respectively characteristic of edge-sharing linkages between Mg and Co octahedra and of corner sharing linkages between Co octahedra and Si tetrahedra, as in clay structures. The angular dependence of the Co-Mg and Co-Si contributions indicates that Co-Mg pairs are oriented parallel to the film plane, whereas Co-Si pairs are not. These results are interpreted by the formation of Co inner-sphere mononuclear surface complexes located at the edges of hectorite platelets, in the continuity of the (Mg, Li) octahedral sheet. Copyright 1999 Academic Press. PMID- 10362484 TI - Synthesis of Platinum Ultrafine Particles in AOT Reverse Micelles. AB - The synthesis of platinum ultrafine particles by the reduction of H2PtCl6 with hydrazine in AOT/isooctane reverse micellar solutions has been studied. By high resolution electron microscope, electron diffraction pattern, and XRD analyses, the resultant particles have been found to be pure platinum of fcc structure. Their sizes were observed to increase with the increases in the molar ratio of water to AOT (omegaO) and in the concentration of H2PtCl6, while they decreased with the increase of hydrazine concentration. At a constant omegaO value, the size of platinum ultrafine particles was not affected significantly when the concentration ratio of hydrazine to H2PtCl6 was above 10, the AOT concentration increased from 0.1 to 0.6 M, and the temperature varied from 15 to 35 degrees C. Furthermore, the kinetic study of particle formation indicated that the nucleation time needed several minutes. The time for the growth of platinum ultrafine particles to their final size after nucleation was about one to several hours. It was observed that the formation rates increased with the increase of omegaO value and the concentrations of AOT and H2PtCl6, but they were not affected by hydrazine concentration when the concentration ratio of hydrazine to H2PtCl6 was above 10. Copyright 1999 Academic Press. PMID- 10362485 TI - XPS Analysis of Carbon Fiber Surfaces-Anodized and Interfacial Effects in Fiber Epoxy Composites. AB - High strength carbon fibers were electrochemically treated in an aqueous ammonium carbonate solution, with increasing electric current density, using an original continuous treatment process. The electrochemical treatment induced a modification of the surface properties, i.e., surface functional groups as observed by XPS. As a result, the O1s/C1s and the N1s/C1s ratios of the fiber surfaces increased with the increase of current intensity of the electrochemical treatment in the interval of about 30-50 A m-2. However, no significant change in the surface functional activity characters was observed for strong treatments. Hence, a moderate treatment (30 A m-2) was sufficient to obtain optimum (O1s + N1s)/C1s ratios in this system. This treatment is possibly suitable for carbon fibers to be incorporated in a polar organic matrix, resulting in increasing the interlaminar shear strength (ILSS) of the resulting composites. Copyright 1999 Academic Press. PMID- 10362486 TI - A Critique of the Mathematical Coherence of Acid/Base Interfacial Free Energy Theory. AB - Acid/base theory has, over the last decade or so, been developed to describe interfacial free energies, or tensions, in wetting theory. An approach put forward by van Oss and co-workers, involving van der Waals/Lifshitz and Lewis electron acceptor/donor contributions to surface/interfacial free energies, has often been employed. The present study considers use of this theory for evaluating surface data for various polymeric surfaces employing known, characterized liquid probes for obtaining contact angle data. Results are analyzed using extended matrix analyses, originally proposed for treating the dispersive/polar interpretation of wetting results, and good agreement with literature values is obtained. By "inverting" the system, i.e., by treating the known solids as probes and rederiving surface data for liquids, inconsistencies are found to arise. Results for wetting of the same polymers and mica, using a two-liquid system (n-octane/water), are exploited to attempt to rederive the surface characteristics of water. Again, serious incoherence is manifest. Despite the conceptual interest of acid/base theory, clearly the mathematical formulation is presently inadequate. Copyright 1999 Academic Press. PMID- 10362487 TI - Synthesis and Properties of Sub-50-nm Europium Oxide Nanoparticles. AB - Eu2O3 nanocrystals are synthesized by a colloidal precipitation route in the size range 2-40 nm. The nanocrystals are passivated with a surface layer of trioctyl phosphine oxide (TOPO) in order to eliminate surface recombination effects. When pumped at 254 nm (4f --> 5d transition) the nanocrystals exhibit red luminescence characteristic of 5D0 --> 7Fn Eu3+ transitions within the cubic form of Eu2O3. The efficiency of the luminescence is increased by a factor of five as the particle size drops below 10 nm; it is suggested that confinement of the long lifetime Eu3+ excitation within the nanocrystal is responsible for this effect. Copyright 1999 Academic Press. PMID- 10362488 TI - Influence of the Level of Cholesteryl Sulfate Present in Stratum Corneum Lipid Liposomes on Their Stability Against Triton X-100. AB - The interaction of Triton X-100 (TX-100) with stratum corneum (SC) lipid liposomes varying the proportion of cholesteryl sulfate (Chol-sulf) was investigated. The surfactant/lipid molar ratios and the bilayer/aqueous phase partition coefficients were determined at sublytic level by monitoring the changes in the fluorescence intensity of liposomes due to the 5(6) carboxyfluorescein released from the interior of vesicles. The fact that the free surfactant concentration was always lower than the surfactant CMC indicates that permeability changes were mainly ruled by the action of surfactant monomers in all cases. The lowest surfactant ability to alter the permeability of SC liposomes and highest surfactant affinity with these bilayer structures was reached when the proportion of Chol-sulf in bilayers was 10%. Futhermore, the highest resistance of SC liposomes to be solubilized by TX-100 (via mixed micelle formation) also occurred at this Chol-sulf proportion, which corresponds to that existing in the intercellular SC lipids. These surfactant effects may be related to the reported dependencies between the level of Chol-sulf in the intercellular lipids and the abnormalities in the skin properties as the barrier function. Copyright 1999 Academic Press. PMID- 10362490 TI - Spontaneous Thermal Convection in the Vicinity of Electrolyte/Water Interfaces. AB - We present theoretical evidence for transient roll-cell convection induced in the vicinity of freshly prepared electrolyte/water interfaces. The mechanism relies on a hitherto unrecognized type of double-diffusive convection. Electrolyte diffusion down the strong concentration gradient at the interface produces local heating just below the interface as the electrolyte becomes more dilute. If the concentration gradient at the interface is sufficiently large, temperature gradients on the order of one Kelvin per centimeter can be induced by this mechanism. We assume the horizontal electrolyte/water interface is contained in a long prism with a square vertical cross section and that the electrolyte solution is denser than the supernatant water layer. Provided that the sidewalls of the prism are thermally conducting, convection commences at the junction of the interface (at half the height of the square section) and these walls. In each vertical semi-cross-section, two roll cells evolve at the interface and the lower cell rapidly gains intensity at the expense of the counter-rotating upper cell. After a short period of time the lower cell migrates diagonally downward to lie just beneath the interface and the associated region of local heating. Although the fluid velocities associated with this roll cell initially increase with time, they do not appear to be sufficiently strong to distort the vertical stratification of the electrolyte's concentration profile. In a cylindrical annulus standing on its circular base the dominant roll cell takes the form of a toroidal vortex resembling a Taylor vortex. Copyright 1999 Academic Press. PMID- 10362489 TI - Magnetite Nanoparticles Prepared by Precipitation from Partially Reduced Ferric Chloride Aqueous Solutions. AB - Spherical magnetite particles with mean diameter of about 10 nm or less were prepared by means of a reduction-precipitation method with ferric chloride as starting material, which was partially reduced to ferrous salts by Na2SO3 before alkalinizing with ammonia. It was found that the nature of the product depends strongly on the initial ratio R0 = [Fe3+]0/[SO2-3]0. The most appropriate ratio was 3 as proved by X-ray diffraction analysis on the samples although the initial concentration of ferric chloride can be different. Particle diameter increased from ca. 3 to ca. 11 nm with an increase of the concentration of aqueous ferric chloride from 0.075 to 0.45 mol dm-3. The advantage of this method lies in the formation of a red intermediate during the reduction process, which enables us to prevent the reoxidation of the ferrous ions by adding precipitation agents at the end of the reduction reaction without the protection of nitrogen or argon. Copyright 1999 Academic Press. PMID- 10362491 TI - Menisci in a Wedge and a Slit for Incomplete Wetting Conditions. AB - This paper presents a closed form analytical solution to the augmented Young Laplace equation for the meniscus profile in two-dimensional wedge- and slit shaped capillaries. The solution is valid for conditions of complete and incomplete wetting and for any form of the disjoining pressure function. Copyright 1999 Academic Press. PMID- 10362492 TI - Determination of a putative phosphate-containing peptide in calreticulin. AB - Calreticulin is an abundant endo/sarcoplasmic reticulum (ER/SR) protein that may carry out multiple functions inside cells. Except for calreticulin, all of the major ER/SR Ca2+-binding proteins are substrates for protein kinase CK2 in vitro, which led us to hypothesize that native calreticulin might exist in the phosphorylated form. To investigate this possibility, we purified calreticulin from cardiac microsomes and verified its identity by immunoblot analysis and sequencing of tryptic peptides. Purified calreticulin, like cardiac calsequestrin, contained endogenous phosphate as determined by a Malachite green assay for phosphate. Previous analyses of cardiac calsequestrin have localized phosphate to a single tryptic peptide containing serine phosphate on sites phosphorylated by protein kinase CK2. Using a similar procedure, we analyzed calreticulin tryptic peptides with Malachite green, localizing phosphate binding to a single calreticulin peptide 367LKEEEEDKK. As this peptide contains no phosphorylatable residues, our results suggest that calreticulin may tightly bind phosphate or a phosphate-containing molecule at this site. PMID- 10362493 TI - Superoxide anion-induced formation of inositol phosphates involves tyrosine kinase activation in smooth muscle cells from rat mesenteric artery. AB - Our previous studies have demonstrated an enhanced production of inositol phosphates (IPs) induced by superoxide in smooth muscle cells (SMCs). The mechanisms for this effect, however, remained largely unknown. In the present study, it was found that superoxide increased IP production in SMCs from rat mesenteric arteries in a time-dependent manner. The effect of superoxide on IP formation was significantly inhibited by the antioxidants n-acetylcysteine or alpha-lipoic acid. Genistein and tyrphostin A25, two tyrosine kinase inhibitors, also inhibited the superoxide-induced IP formation. The application of monoclonal antibody against phospholipase Cgamma (PLCgamma) significantly inhibited the superoxide-induced IP formation. Finally, the expression level of PLCgamma proteins was increased 6 hrs after exposing SMCs to superoxide. The present findings demonstrate that superoxide activates the tyrosine kinase pathway and suggest that the tyrosine kinase-mediated IP formation may represent a novel mechanism underlying the signalling role of superoxide in rat mesenteric artery SMCs. PMID- 10362494 TI - Selenocysteine-containing thioredoxin reductase in C. elegans. AB - Mammalian thioredoxin reductases contain a TGA-encoded C-terminal penultimate selenocysteine (Sec) residue, and show little homology to bacterial, yeast, and plant thioredoxin reductases. Here we show that the nematode, Caenorhabditis elegans, contains two homologs related to the mammalian thioredoxin reductase family. The gene for one of these homologs contains a cysteine codon in place of TGA, and its product, designated TR-S, was previously suggested to function as thioredoxin reductase. The other gene contains TGA and its product is designated TR-Se. This Sec-containing thioredoxin reductase lacks a canonical Sec insertion sequence element in the 3'-untranslated area of the gene. TR-Se shows greater sequence similarity to mammalian thioredoxin reductase isozymes TR1 and TR2, whereas TR-S is more similar to TR3. TR-Se was identified as a thioredoxin reductase selenoprotein by labeling C. elegans with 75Se and characterizing the resulting 75Se-labeled protein by affinity and other column chromatography and gel-electrophoresis. TR-Se was expressed in Escherichia coli as a selenoprotein when a bacterial SECIS element was introduced downstream of the Sec TGA codon. The data show that TR-Se is the major naturally occurring selenoprotein in C. elegans, and suggest an important role for selenium and the thioredoxin system in this organism. PMID- 10362495 TI - M-type 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase is the product of a late muscle differentiation gene. AB - Genes that are expressed in adult muscle, but not in myotubes, are useful markers of the last steps of muscle maturation. We have investigated at what stage of differentiation the muscle-specific (M) promoter of a gene that codes for 6 phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFK2) becomes functional. M PFK2 mRNA, which is present in adult muscle, did not appear during differentiation of L6 myoblasts into myotubes induced by growth factor withdrawal and hormonal treatment, even when this differentiation was stimulated by expression of transgenes coding for myf-5 or Rb. A comparison with the expression pattern of muscle genes showed that M-PFK2 is a marker of mature skeletal muscle. We also found that M-PFK2 is expressed in both types (slow-twitch and fast twitch) of adult muscle. Thus, the M-PFK2 promoter is a novel model for studying the transcriptional control of the final steps of muscle differentiation that are common to the two types of myofibers. PMID- 10362496 TI - Subclass specificity of autoantibodies against myosin in patients with idiopathic dilated cardiomyopathy: pro-inflammatory antibodies in DCM patients. AB - Detection of antimyosin antibodies in non-inflammatory cardiac disease undermines their disease specificity as a sensitive marker of damage in dilated cardiomyopathy (DCM) patients. Antibody subclass specificity could provide a more sensitive marker of disease and possibly discriminate the humoral autoimmune responses in different cardiac diseases. Frequency and reactivity of autoantibodies against alpha- and beta-isoforms of myosin heavy chain (mhc) were evaluated by ELISA for IgG, IgM, and subclasses IgG1, IgG2, and IgG3 in patients with DCM (NYHA III/IV, n = 82), end stage ischemic heart disease (E-IHD: NYHA III/IV, n = 62), mild ischemic heart disease (NYHA I/II, n = 27), and controls (n = 54). Autoantibodies against atrial and ventricular myosin were raised in heart failure patients compared to mild-IHD and controls but with different antigen affinities. Reactivity in E-IHD was significantly raised against (ventricular) beta-mhc compared with only mild-IHD patients, suggesting a relative increase in ventricular specific antibodies in IHD patients with a higher NYHA class. IgG subclass analysis for IgG1, IgG2, and IgG3 against alpha- and beta-mhc showed statistically raised levels of IgG3 only in DCM patients and a significantly higher reactivity of IgG2 in heart failure patients versus controls. The results demonstrate immunological heterogeneity of antimyosin antibodies developed in different clinical entities. Pro-inflammatory characteristics of IgG3 antibodies in a select group of patients with DCM may contribute to autoimmune mechanisms of injury in these patients. PMID- 10362497 TI - Characterization of a 3'-5' exonuclease associated with VDJP. AB - VDJP (V(D)J RSS Dependent DNA Joining Protein) was cloned based on binding to the nonamer portion of the V(D)J recombinational signal sequence (RSS), and genetic analysis revealed that VDJP is encoded by the same gene as the large subunit of Replication Factor C (RF-C). Recombinant VDJP has a site directed DNA joining activity and is capable of forming a covalent bond between DNA fragments containing an RSS element near their ends and exhibits 3' to 5' exonuclease activity. In this report, we examine the biochemical properties of the VDJP exonuclease activity such as directionality of nuclease action (3' to 5' or 5' to 3'), single-strand substrate preference, cleavage products, dependence on cofactors and metal cations, and optimal reaction conditions. From this analysis, we conclude that VDJP has an intrinsic 3'-5' exonuclease activity that produces mononucleotide products. PMID- 10362498 TI - Mass spectrometric determination of a novel modification of the N-terminus of histidine-tagged proteins expressed in bacteria. AB - Two proteins, FKBP, and Spo0F, were expressed in bacteria as histidine-tagged fusion proteins and isolated under native conditions. MALDI-TOF-MS analysis revealed that each protein preparation contained two components, neither of which corresponded to the molecular weights predicted from DNA sequences. The difference in molecular weight between the two FKBP components and two Spo0F components was approximately 178 +/- 14 Da. Site-specific proteolytic cleavage resulted in the release of histidine-tagged peptide from the recombinant proteins. MALDI mass spectra of the cleaved proteins showed a single molecular ion peak for each species with the predicted molecular weights. The histidine tagged peptide released from both fusion proteins displayed two distinct peaks by MALDI-FT-MS corresponding to monoisotopic molecular weights of 2269. 027 Da and 2447.087 Da, respectively, which were both inconsistent with the predicted peptide sequence M-G-H-H-H-H-H-H-H-H-H-H-S-S-G-H-I-E-G-R of 2400.055 Da. The peptide at 2269.027 Da was sequenced by ESI-MS-MS and found to be a truncated histidine tag resulting from an initiator methionine deletion. ESI-MS-MS analysis of the peptide at 2447.087 Da indicated a moiety of 178.0 Da attached to the second residue glycine of the histidine tag. This alteration of the N-terminus does not fit any known modifications. A synthetic peptide with the identical sequence of the isolated his-tag M-G-H-H-H-H-H-H-H-H-H-H remained unmodified during the protein purification process, suggesting that modification of the initiator methionine was carried out in vivo, rather than the result of a chemical reaction from the isolation procedure. PMID- 10362499 TI - Calcium-induced p56(Lck) phosphorylation in human T lymphocytes via calmodulin dependent kinase. AB - We report that stimulation of both primary human and Jurkat T lymphocytes with the calcium ionophore ionomycin, or A23187, results in the phosphorylation of p56(Lck) as determined by shifts in mobility of p56(Lck) on immunoblots. The shifts in the mobility of p56(Lck) induced by ionomycin could be blocked by preincubation of the cells with EGTA, demonstrating the requirement for extracellular calcium in this response. Although increases in intracellular calcium have been shown to modulate CD45 activity, phosphorylation of p56(Lck) was not mediated via CD45. Ionomycin stimulation of J45. 01 cells, a CD45 negative Jurkat cell derivative, also resulted in p56(Lck) mobility shifts. Instead, this response appears to be mediated via a calmodulin-dependent kinase. This response could be blocked by calmidazolium, an inhibitor of calmodulin, and KN-93, an inhibitor of calmodulin-dependent kinases (CaM-Kinase). KN-92, an inactive analog of KN-93, failed to block this response. These studies demonstrate a new role for calcium and CaM-Kinase in human T-lymphocytes and describe a novel mechanism by which p56(Lck) can be modulated. PMID- 10362500 TI - Engagement of spectrin and actin in capping of FcgammaRII revealed by studies on permeabilized U937 cells. AB - Plasma membrane receptors can undergo translocation in the plane of plasma membrane after binding of polyvalent ligands. Ligand/receptor clusters, named patches, can collect into a polar cap, presumably due to their association with the submembrane actin-based cytoskeleton. We found that the assembly of Fcgamma receptor II caps in human monocytic U937 cells was accompanied by the accumulation of spectrin and actin in the cap region. Permeabilization of cells with streptolysin O rendered capping sensitive to inhibition by phalloidin, an actin filament stabilizing agent. A rabbit antibody directed against the chicken erythrocyte alpha-subunit of spectrin, an actin- and membrane-binding protein, also blocked the capping in a dose dependent manner. The inhibition reached approximately 50% after 20 minutes of cell treatment with the antibody. Anti alpha-spectrin targeted specifically its submembrane antigen, in contrast to unspecific antibodies which remained dispersed in the cell interior and had no influence on the cap assembly. Our results indicate an active engagement of spectrin and actin filaments in the capping of Fcgamma receptor II. PMID- 10362501 TI - Unique features of dendritic cells in IFN-gamma transgenic mice: relevance to cancer development and therapeutic implications. AB - Although induction of interferon gamma (IFN-gamma) and activation of antigen presenting dendritic cells (DCs) are two vital events during an immune response, the impact of endogenous IFN-gamma on DC function has yet to be clarified. The phenotype and function of DCs isolated from mice with high (IFN-gamma-transgenic mouse [Tg]) and undetectable levels of circulating IFN-gamma (normal mice [NM]) were therefore compared. The capacity to stimulate allogenic (p < 0.05) and antigen-specific T lymphocytes (p < 0.05), as well as the ability to produce IL 12 (p < 0.05) and to process soluble protein antigens (p < 0.05) was found to be significantly higher in DCs from the Tg mice compared to the NM case. The presence of activated DCs in a microenvironment of endogenous IFN-gamma suggests that the IFN-gamma-Tg mouse is a suitable animal model to study cancer immunotherapy in vivo. PMID- 10362502 TI - Role of the 78-kDa glucose-regulated protein as an activity modulator of protein phosphatase1gamma2. AB - We have previously found the 78-kDa glucose-regulated protein (Grp78) to be a subunit of protein phosphatase1(PP1)gamma2. To determine the role of Grp78 in PP1gamma2 holoenzyme, we compared the two forms of this enzyme, PP1gamma2 holoenzyme containing Grp78 and Grp78-dissociated PP1gamma2 in rat testes in terms of their kinetic constants and sensitivities to inhibitors of this enzyme. The enzymatic activity of the Grp78-dissociated enzyme was much lower at whole range of concentrations of a substrate (phosphorylase a) than that of the holoenzyme; the Km value was about ten-fold higher in Grp78-dissociated enzyme than in holoenzyme, while the Vmax was similar. IC50s of the Grp78-dissociated enzyme for three inhibitors (microcystin-LR, inhibitor-2, and okadaic acid) were more than ten-fold higher than those of the holoenzyme. These results indicate that the Grp78 subunit modulates the activity of PP1gamma2 through its actions to control the binding of substrates or inhibitors to PP1gamma2. PMID- 10362504 TI - Detection of apolipoprotein B mRNA editing by peptide nucleic acid mediated PCR clamping. AB - Apolipoprotein B (apoB) mRNA editing leads to a single base change in its mRNA and the production of apoB-48. Currently, the degree of apoB mRNA editing is analyzed by the RT-PCR primer extension method. While this method is quantitative, it is labor intensive, utilizes radioactivity for labeling and may not be sensitive enough to discriminate between low levels of editing and inherent assay background levels. Peptide nucleic acid (PNA) oligonucletides have been used in single point mutation detection through PCR clamping. In the present work, we developed a PCR based assay which can detect the single base change responsible for the apoB-48 production. We found that as low as 0.5% of the edited form can be clearly detected by PNA mediated PCR clamping. When combined with the primer extension assay, an approximately 180-fold enrichment of the edited percentage is observed, reflecting selected PCR amplification of templates containing the edited base. PMID- 10362503 TI - Cloning of a cDNA encoding a novel cytochrome P450 from the insect Locusta migratoria: CYP6H1, a putative ecdysone 20-hydroxylase. AB - The biosynthesis of the steroidal molting hormone, 20-hydroxyecdysone, of arthropods involves a series of cytochrome P450-catalyzed hydroxylations. None of the many sequences of insect cytochromes P450, known to date, is related to ecdysteroid pathways. Here, we report the cloning and sequencing of a full-length cDNA of a new cytochrome P450, classified as CYP6H1, from malpighian tubules of the locust, Locusta migratoria. The 1854 bp DNA contained an open reading frame coding for a protein of 542 amino acids, a 5'-leader sequence and a 3' untranslated region containing a polyadenylation signal and a poly(A) tail. The encoded protein had been isolated as an ecdysone-binding cytochrome P450 from microsomes of the same tissue in previous work. The closest homolog of CYP6H1 was CYP6A2 from Drosophila with 42.1% identity. According to Northern analysis, CYP6H1 is predominantly expressed at larval instars and in malpighian tubules. Evidence is presented for a functional assignment of CYP6H1 to microsomal ecdysone 20-hydroxylase of the locust. PMID- 10362505 TI - Monocytic differentiation of HL-60 cells is characterized by the nuclear translocation of phosphatidylinositol 3-kinase and of definite phosphatidylinositol-specific phospholipase C isoforms. AB - Immunochemical and immunocytochemical data indicate that nuclei of HL-60 cells contain different enzymes involved in the phosphoinositide cycle, such as PI 3-K and the phosphatidylinositol-specific PLC isoforms beta3, gamma1 and gamma2. These enzymes translocate differently to the nuclear fraction when HL-60 cells are treated with differentiating doses of vitamin D3: PI 3-K translocated progressively to the nucleus in parallel with full differentiation until 96 hours. PLC beta3 increased until 72 hours of treatment and then lowered its intranuclear amount and PLC gamma1 was unchanged at all the examined times. PLC gamma2 nuclear translocation increased progressively until 96 hours of vitamin D3 administration. A fourth PLC isozyme, beta2, present in the cytoplasm of untreated cells, translocates to the cytoplasm after vitamin D3 addition and reaches the highest concentration at the end of monocytic differentiation. Terminal monocytic differentiation was characterized at the nuclear level by high levels of PI 3-K and PLC gamma2 and by the novel expression of PLC beta2. We then observed that the xi isoform of PKC, constitutively present in nuclei of HL-60 cells, translocated to the nucleus when cells were induced to differentiate along the monocytic lineage, but the nuclear translocation of PKC xi was blocked as a consequence of PI 3-K inhibition by Wortmannin. These findings indicate that the main components of the noncanonical and canonical inositol lipid signal transduction pathways, including PI 3-K, PLC beta2 and beta3, PLC gamma2, undergo nuclear translocation and may therefore play a relevant role during monocytic differentiation at the nuclear level. Furthermore, PKC xi nuclear translocation appears to be related to PI 3-K activity. PMID- 10362506 TI - Stimulation of prolactin release by prolactin-releasing peptide in rats. AB - We have previously reported a hypothalamic peptide that shows specific prolactin (PRL)-releasing activity in vitro, named prolactin-releasing peptide (PrRP). However, its activity in vivo has not yet been shown. In this study, we examined whether PrRP could induce specific PRL release in vivo using normal cycling female and male rats. Intravenous injection of PrRP31 increased plasma PRL levels in rats in a dose-dependent manner. PrRP31 (50 nmol/kg i.v.) significantly (P < 0.05) stimulated plasma PRL levels within 25 min after injection in rats in proestrus, estrus, and metestrus. A higher dose of PrRP31 (500 nmol/kg i.v.) was necessary for a significant increase in plasma PRL levels in male rats. These results clearly indicate that female rats, especially at proestrus, are more sensitive to PrRP-induced PRL secretion than male rats. The effect of PrRP on PRL release is affected considerably by the estrous cycle and sex, which suggests that PrRP sensitivity is controlled by the endogenous hormonal milieu, such as estrogen levels. PrRP31 did not affect other pituitary hormone secretions. The results indicate that PrRP shows specific PRL-releasing activity in vivo as well as in vitro and suggest that it plays an important role in the regulation of PRL release under certain physiological conditions. PMID- 10362507 TI - Modulation of cytochrome c-mediated extramitochondrial NADH oxidation by contact site density. AB - Data presented in previous reports suggest that in rat liver mitochondria a "bi trans-membrane" electron transport pathway is present which promotes the transfer of reducing equivalents directly from cytosolic NADH to molecular oxygen inside the mitochondria. Here we show that the oxidation of external NADH is stimulated by atractylate + ADP and greatly inhibited by glycerol. These two conditions have been documented to promote the increase and the decrease respectively of the frequency of "contact sites" between the two mitochondrial membranes. NADH oxidation is not affected at all by glycerol and atractylate + ADP when TMPD and endogenous cytochrome c are utilized as electron carriers. The results obtained are consistent with the proposal that the bi-trans-membrane electron transport chain might be localized at the level of respiratory contact sites having the function of promoting the oxidation of the surplus amount of cytosolic NADH. This electron transport pathway has been suggested to play a decisive role in the early stages of apoptosis [Biochem. Biophys. Res. Commun. 246, 556-561, 1998]. PMID- 10362508 TI - Detection of multiple patterns of oscillatory oxygen consumption in single mouse islets of Langerhans. AB - A novel oxygen microsensor was used to measure oxygen levels in single mouse islets as a function of glucose concentration. Oxygen consumption of individual islets was 5.99 +/- 1.17, 9.21 +/- 2.15, and 12.22 +/- 2.16 pmol/min at 3, 10, and 20 mM glucose, respectively (mean +/- SEM, n = 10). Consumption of oxygen was islet-size dependent as larger islets consumed more oxygen than smaller islets but smaller islets consumed more oxygen per unit volume than larger islets. Elevating glucose levels from 3 to 10 mM induced pronounced fast oscillations in oxygen level (period of 12.1 +/- 1.7 s, n = 6) superimposed on top of large slow oscillations (period of 3.3 +/- 0.6 min, n = 6). The fast oscillations could be completely abolished by treatment with the L-type Ca2+-channel blocker nifedipine (40 microM) with a lesser effect on slow oscillations. Slow oscillations were almost completely dependent upon extracellular Ca2+. The oxygen patterns closely mimic those that have previously been reported for intracellular Ca2+ levels and are suggestive of an important role for Ca2+ in amplifying metabolic oscillations. PMID- 10362509 TI - Clusterin prevents aggregation of neuropeptide 106-126 in vitro. AB - The prion/amyloid neuropeptide 106-126 spontaneously aggregates to form fibrillar structures in vitro. The aggregation in vitro could be prevented in a dose related manner by clusterin, and the specificity of this action was confirmed by reversal with antibody to clusterin. The relevance of these observations is discussed in relation to previous observations that clusterin and PrPBSE colocalise in naturally occurring cases of BSE. PMID- 10362510 TI - Platelet surface membranes are highly mitogenic for coronary artery smooth muscle cells--A novel mechanism for sustained proliferation after vessel injury? AB - The effects of isolated platelet surface membranes on DNA synthesis and proliferation of bovine coronary artery smooth muscle cells (SMC) were studied. Platelet membranes were very potent mitogens for SMC. The potency was about 10 fold higher than the maximum effects of platelet-derived growth factor-BB (PDGF). Platelet membrane-induced mitogenesis was inhibited by rapamycin, wortmannin or heating for 15 min at 70 degrees C but not by the PDGF receptor antagonist SCH 13.929 or by neutralizing PDGF antibodies. Only a partial (30%) inhibition was seen with PD 98059. In contrast, PDGF-induced SMC mitogenesis was heat-stable but sensitive to SCH 13. 929, PDGF antibodies, and PD 98059. These findings provide evidence for a novel mechanism for platelet-induced SMC proliferation that is independent of PDGF secretion. Platelet membranes, attached to or incorporated into the vessel wall, could maintain sustained SMC proliferation following injury. PMID- 10362511 TI - A novel polymorphism in the MCP-1 gene regulatory region that influences MCP-1 expression. AB - Two novel polymorphisms in the distal regulatory region of the MCP-1 gene were identified by directly sequencing PCR amplified genomic DNA. These polymorphisms are located at positions -2518 (G or A) and -2076 (A or T) relative to the major transcriptional start site of the gene. To examine the effect of these polymorphisms on MCP-1 transcription, polymorphic variants of the MCP-1 distal regulatory region were placed upstream of a luciferase reporter gene and transfected into A172 cells. IL-1beta-induced luciferase activity was significantly greater from cells transfected with constructs containing G at position -2518. The polymorphism at -2076 did not affect MCP-1 transcription. IL 1beta-treated peripheral blood mononuclear cells from individuals heterozygous or homozygous for G at -2518 produced more MCP-1 than cells from individuals homozygous for A at -2518. These data identify a polymorphism in the MCP-1 distal regulatory region that affects the level of MCP-1 expression in response to an inflammatory stimulus. PMID- 10362512 TI - A novel gene (oculomedin) induced by mechanical stretching in human trabecular cells of the eye. AB - To understand molecular mechanisms underlying the response to pressure in human trabecular cells of the eye, genes induced by cyclic mechanical stretching were isolated by subtractive hybridization assisted by polymerase chain reaction. A novel gene containing an Alu repetitive element in the 5' untranslated region was identified, and its expression was confirmed by Northern blot analysis to be stretch-specific in trabecular cells. The gene was also expressed in the retina, but not in the other tissues, including the brain. The gene encoded a putative small protein with 44 amino acids, which showed homology with neuromedin K. The putative novel protein was named as "oculomedin," and would be used as a candidate gene for glaucoma. PMID- 10362513 TI - Selective oxidation of methionine residues in prion proteins. AB - Prion proteins are central to the pathogenesis of several neurodegenerative diseases through the postulated conversion of the endogenous cellular isoform (PrPc) into a pathogenic isoform (PrPSc). Although the cellular function of normal prion protein remains unresolved a number of studies have shown that prion proteins may be involved in the cellular response to oxidative stress. Here, using purified recombinant sources of mouse and chicken PrP refolded in the presence of copper (II) we show that the methionine residues of the protein are uniquely susceptible to oxidation. We suggest that Met residues may form an essential part of the mechanism of the antioxidant activity exhibited by normal prion protein. PMID- 10362514 TI - Identification of phosphorylation sites in the PKD1-encoded protein C-terminal domain. AB - The PKD1-encoded protein, "polycystin-1", has a large N-terminal extracellular portion, multiple transmembrane domains, and a short intracellular C-terminal tail with four tyrosine residues and two putative sites for serine phosphorylation. Its function in kidney development and autosomal dominant polycystic kidney disease (ADPKD) is still unknown. We have subcloned the cDNA encoding the polycystin-1 C-terminal domain (PKD1-CTD) into a prokaryotic expression vector, and site-directed mutagenesis was performed to target the four tyrosine residues and four serine residues in two putative phosphorylation sites. In vitro phosphorylation assays were conducted on both wild type and mutant PKD1 CTD fusion proteins. It was found that the wild type PKD1-CTD and all mutant fusion proteins, except S4251G/S4252G, could be phosphorylated by lysates from cultured normal human renal collecting tubule (NHCT) cells, as well as by commercially purified cAMP-dependent protein kinase (PKA). The phosphorylation of the PKD1-CTD fusion protein by NHCT lysates was greatly enhanced by cAMP and its analog 8-Br-cAMP, and inhibited by the specific PKA inhibitors PKI(6-22) and H 89. Activators and inhibitors of protein kinase C (PKC) had no effects on the phosphorylation of the PKD1-CTD fusion protein. Using commercially purified pp60(c-src) (c-src) it was also shown that the PKD1-CTD fusion protein could be phosphorylated by c-src in vitro, and that this phosphorylation could be abolished by a mutation Y4237F. By comparing the amino acid sequence at 4249-4253 (RRSSR) with the consensus sequence for PKA phosphorylation (RRXSX), we suggest that the serine residue at 4252 is the target of phosphorylation by a cAMP dependent protein kinase in NHCT cell lysates. In addition, we suggest that Y4237 might be phosphorylated by c-src in living cells. PMID- 10362515 TI - Synergistic activation of the human Btk promoter by transcription factors Sp1/3 and PU.1. AB - Analysis of the human Bruton's agammaglobulinemia tyrosine kinase (Btk) gene promoter revealed that 280 bp upstream of the transcriptional start site is sufficient for a cell restricted expression pattern. Here, the interplay of the transcription factors Sp1, Sp3, and PU.1 binding to this promoter area was analysed. All three proteins are able to independently activate the promoter in Drosophila Schneider (SL2) cells lacking endogenous Sp- or PU.1-like activities. Furthermore, PU.1 is able to act synergistically with Sp1 as well as Sp3 to transactivate the promoter. This transactivation is mediated through adjacent binding sites rather than through the more distant Sp binding site, suggesting a possible direct interaction between PU.1 and Sp1/3. Expression of Btk was found in ES cells and levels of expression were the same as in ES cells with a targeted deletion of the Sp1 gene, suggesting that Sp3 acts as a positive regulator of Btk in vivo, in the absence of Sp1. PMID- 10362516 TI - Activity in monomers of human cytomegalovirus protease. AB - Herpesvirus proteases require dimerization for activity, although crystallographic data indicate that each monomeric subunit possesses a well separated and complete active site. This suggests that dimerization stabilizes the monomeric protease subunits in an active conformation. Chemical cross-linking with disuccinimidyl glutarate was used to capture human cytomegalovirus protease in its various conformations. The cross-linked protease retained activity under mildly chaotropic conditions (0.25 to 0.75 M urea) in contrast to non-cross linked protease which lost activity. Identification of active protease species by incorporation of radioactive diisopropylfluorophosphate showed that in addition to cross-linked dimers, cross-linked protease monomers were responsible for a significant fraction of the total protease activity. These results are consistent with the hypothesis that herpesvirus protease activation occurs by stabilization of an active conformer in the dimer. PMID- 10362517 TI - Discrepancies in the measurement of UVC-induced 8-oxo-2'-deoxyguanosine: implications for the analysis of oxidative DNA damage. AB - Ultraviolet (UV) light-induced indirect, oxidative damage to DNA has received increasing attention with respect to the mutagenic and carcinogenic effects of solar radiation. An oxidative lesion that has raised particular interest because of its qualitative and quantitative importance is 8-oxo-2'-deoxyguanosine. This deoxynucleoside lesion is most frequently measured by high performance liquid chromatography with electrochemical detection (HPLC-EC) following enzymatic hydrolysis of DNA or as the base equivalent, 8-oxoguanine, by gas chromatography mass spectrometry (GC-MS) following acid hydrolysis of DNA. We have noted a discrepancy in the literature whereby the levels of 8-oxo-2'-deoxyguanosine measured by HPLC-EC in UVC-irradiated DNA are significantly higher than when 8 oxoguanine is measured by GC-MS. By making use of the availability of both HPLC EC and stable-isotope dilution GC-MS methodologies in our laboratory we have confirmed the discrepancy noted in the literature by parallel analysis of the same UVC-irradiated calf thymus DNA samples. Furthermore, analysis of the UVC induced product by UV-visible spectrophotometry, voltammetry and its detection by a monoclonal antibody which recognises 8-oxo-2'-deoxyguanosine strongly suggests that the product is indeed 8-oxo-2'-deoxyguanosine. Partial explanation for this discrepancy could be an inordinate resistance of UVC-irradiated DNA to formic acid hydrolysis. However, we cannot completely exclude the possibility that there is a formic acid-labile species which co-elutes with 8-oxo-2'-deoxyguanosine in enzymatically digested UVC-irradiated DNA. Whether this phenomenon is unique to UV-irradiation damage or occurs with other systems that cause oxidative damage to DNA awaits further investigation. Irrespective of the exact mechanism, there will be significant implications for the analysis of oxidative DNA damage. PMID- 10362518 TI - Antiproliferative and GH-inhibitory activity of chimeric peptides consisting of GHRP-6 and somatostatin. AB - Chimeric peptides consisting of growth hormone releasing peptide (GHRP-6) linked to somatostatin (6-11) via an amide bond to provide the effector parts of both the peptides were synthesized. The anti-proliferative, cytotoxic, and GH inhibitory activities of these chimeric peptides were determined in vitro in the rat pituitary adenoma cell line GH3. One of the chimeric peptides, GSD, exhibited significantly greater (p < 0.001) anti-neoplastic and GH-inhibitory activity, as compared to RC-160. The hybrid peptides displayed high affinity binding to somatostatin receptors on GH3 cells. The bioactivity of GSD was found to be mediated by the stimulation of tyrosine phosphatase, involving a cGMP-dependent pathway, through pertussis toxin-sensitive G-proteins. Such potent GH-inhibitory chimeric peptides may be of potential importance in the therapy of acromegaly, as well as provide novel tools to study the regulation of GH secretion by GHRP and somatostatin. PMID- 10362519 TI - Two divergent signaling pathways for TGF-beta separated by a mutation of its type II receptor gene. AB - Transforming growth factor beta (TGF-beta) can inhibit epithelial cell growth and induce extracellular matrix formation through signal transduction via its two receptors and its downstream intracellular Smad proteins. We recently reported a germline mutation, i.e., substitution of methionine for threonine at codon 315 in the kinase subdomain IV, of the TGF-beta type II receptor gene in a kindred of hereditary nonpolyposis colorectal cancer without microsatellite instability and found that the mutant receptor abolished the signal transduction for growth inhibition by TGF-beta. In this study, we performed further functional analysis of this mutant receptor. The results showed that, in contrast to its failure to mediate growth inhibition by TGF-beta, the mutant receptor still retained the ability to induce one of the extracellular matrix proteins, plasminogen activator inhibitor type 1, upon TGF-beta treatment. However, coincident with its failure to mediate growth inhibition by TGF-beta, the mutant receptor failed to transcriptionally upregulate one of the cyclin-dependent kinase inhibitors, p15(INK4B), in response to TGF-beta. These data suggest that threonine 315 of the TGF-beta type II receptor is dispensable for extracellular matrix protein production, but is essential for the growth inhibition by TGF-beta, and that the lack of growth inhibition due to the mutant receptor is possibly mediated through its failure to upregulate p15(INK4B). PMID- 10362520 TI - RNase treatment of yeast and mammalian cell extracts affects in vitro substrate methylation by type I protein arginine N-methyltransferases. AB - Type I protein arginine N-methyltransferases catalyze the formation of omega-NG monomethylarginine and asymmetric omega-NG, NG-dimethylarginine residues using S adenosyl-l-methionine as the methyl donor. In vitro these enzymes can modify a number of soluble methyl-accepting substrates in yeast and mammalian cell extracts including several species that interact with RNA. We treated normal and hypomethylated Saccharomyces cerevisiae and RAT1 cell extracts with RNase prior to in vitro methylation by recombinant protein N-arginine methyltransferases and found that the methylation of certain polypeptides is enhanced up to 12-fold whereas that of others is diminished. 2-D gel electrophoresis of RNase-treated yeast extracts allowed us to tentatively identify the glycine- and arginine-rich (GAR) domain-containing proteins Gar1, Nop1, Sbp1, and Npl3 as major methyl acceptors based on their known isoelectric points and apparent molecular weights. These results suggest that the methylation and RNA-binding of GAR domain containing proteins in vivo may regulate protein-nucleic acid or protein-protein interactions. PMID- 10362521 TI - Human cathepsins F and W: A new subgroup of cathepsins. AB - Human cathepsin F is a recently described papain-like cysteine protease of unknown function. To investigate the evolutionary relatedness to other human cathepsins, we determined the genomic organization and the chromosomal localization of cathepsin F and isolated its putative promoter region. The gene of human cathepsin F (CTSF) is composed of twelve exons and eleven introns and was found to be similar to that of cathepsin W but different from the cathepsins K, S, L, O, B, and C. The splice sites of nine out of the eleven introns were identical to those determined in the cathepsin W gene (CTSW), whereas introns one and ten were unique for CTSF. The 4. 7 kb gene was mapped to the long arm of chromosome 11 at position q13.1-3, a locus shared with CTSW. Phylogenetic analysis of human cathepsin protein sequences demonstrated that (i) cathepsins F and W are evolutionarily separated from other human cathepsins, and (ii) cysteine proteases closely related to human cathepsin W and F are also expressed in parasites and mammals. Based on these phylogenetic findings, on the presence of a particular protein motif ("ERFNAQ") in the propeptides of cathepsins F and W as well as the genomic organization and chromosomal localization of their genes, we concluded that F and W form a novel subgroup of cathepsin proteases. We suggest the naming "cathepsin F-like" proteases distinct from the previously described cathepsins "L- and B-like" subgroups. PMID- 10362522 TI - Precise identification of gene products in hearts after in vivo gene transfection, using Sendai virus-coated proteoliposomes. AB - Both efficient gene transfer and the exact identification of gene product are required for gene therapy. Gene transfection of green fluorescence protein (GFP) might be useful for the reporter. After in vivo cotransfection of GFP and beta galactosidase (beta-Gal) genes in Sendai virus-coated proteoliposomes to rat hearts, we compared the sensitivity and specificity of three methods: GFP detection, histochemical staining (HC) of beta-Gal activity, and immunostaining (IS) of the beta-Gal protein. Fluorescence microscopy and double staining of HC and IS revealed that both GFP and IS were equally sensitive and fourfold superior to HC at the peak of gene expression. However, different from skeletal muscle, the GFP of transfected cardiomyocytes showed two demerits: the fluorescence quenching due to the intense staining of beta-Gal activity, and nonspecific autofluorescence from myocardium. Thus, specific IS would be so far the most reliable to identify the gene product in heart. PMID- 10362524 TI - A model of the complex between cyclin-dependent kinase 5 and the activation domain of neuronal Cdk5 activator. AB - Tau protein kinase II (TPKII) is a heterodimer comprising a catalytic cyclin dependent kinase subunit (Cdk5) and a regulatory protein called neuronal Cdk5 activator (Nck5a). TPKII is somewhat reminiscent, therefore, of the Cdk2-cyclin complex important in cell cycle regulation. In fact, although the amino acid sequence of Nck5a has little similarity to those of cyclins, recent experimental results obtained by site-directed mutagenesis studies have indicated that its activation domain, Nck5a*, may adopt a conformation of the cyclin-fold structure. Based on this structural inference, a 3-dimensional model of the Cdk5-Nck5a*-ATP complex was derived from the X-ray structure of Cdk2-cyclinA-ATP complex. The computed structure for TPKII is fully compatible with experimental data derived from studies of the Cdk5-Nck5a system, and also predicts which amino acid residues might be involved in formation of the Cdk5-Nck5a* interface and ATP binding pocket in TPKII. The computational structure also shows the interactive region of Nck5a* and the T-loop of Cdk5, a critical region in TPKII which functions as a gate-control-lever of the catalytic cleft. Furthermore, a physical mechanism is put forth to explain why the activation of TPKII is not dependent upon phosphorylation of the Cdk5 subunit, a puzzle long-standing in this area. These findings provide a model with which to consider design of compounds which might serve as inhibitors of TPKII. PMID- 10362523 TI - A HMG-CoA reductase inhibitor improved regression of atherosclerosis in the rabbit aorta without affecting serum lipid levels: possible relevance of up regulation of endothelial NO synthase mRNA. AB - We determined the role of Fluvastatin: HMG-CoA reductase inhibitor on the regression of atherosclerosis following removal of dietary cholesterol. Male rabbits fed a 0.5% cholesterol diet for 12 weeks were divided into three groups: A1, hypercholesterolemic; A2, fed a regular diet for an 12 additional weeks; and A3, fed a regular diet with fluvastatin (2 mg/kg/day). Fluvastatin treatment (A3) did not affect serum lipid levels compared with A2. However, it decreased the atherosclerotic area in the aortic arch and decreased total and esterified cholesterol concentrations in the descending aorta. Tone-related basal NO release in the thoracic aorta was larger in A3 than in A2. eNOS mRNA in vessel was determined by competitive RT-PCR assay. It increased in A1, compared with normal aorta and decreased in A2; however, it did not decrease in A3. This is the first report of a decrease in eNOS mRNA in atherosclerosis after removal of dietary cholesterol and a reversal of it by a HMG-CoA reductase inhibitor, which may contribute to the regression of atherosclerosis. PMID- 10362525 TI - Alteration of hMSH2 and DNA polymerase beta genes in breast carcinomas and fibroadenomas. AB - Genomic stability is preserved by error-free DNA replication, post-replicative proofreading, DNA repair, and recombinational events. In essence, DNA repair genes are recognized to play key roles in such stability. We report evidence for expression of the wild-type and a truncated form of DNA polymerase beta (polbeta) proteins, a base-excision repair gene, in breast carcinomas and fibroadenomas, a benign breast disease. An 87-bp deleted variant of polbeta was identified to be prevalent in microsatellite unstable breast tumors and fibroadenomas. A large deletion of 1476 bp, as well as point mutations in human MutS homolog 2 (hMSH2) cDNA, was revealed in breast carcinomas. The protein truncation assay confirmed the 1476-bp deletion as a premature protein. This is the first evidence for variant forms of hMSH2 that are associated with breast cancer. Genomic instability in the hMSH2 and polbeta genes may facilitate the occurrence of mutator phenotype in breast cancer. PMID- 10362526 TI - Posttranscriptional regulation of mammalian methionine synthase by B12. AB - Methionine synthase is one of two key enzymes involved in the removal of the metabolite, homocysteine. Elevated homocysteine levels constitute a risk factor for cardiovascular diseases and for neural tube defects. In cell culture, the activity of methionine synthase is enhanced several-fold by supplementation with its cofactor, B12. The mechanism of this regulation is unknown, although it has been ascribed to a shift from apoenzyme to holoenzyme. Using sensitive assay techniques as well as a combination of Northern and Western analyses, we demonstrate that the effect of B12 on induction of methionine synthase activity is paralleled by an increase in the level of the enzyme. These studies exclude conversion of apoenzyme to holoenzyme as a basis for activation that had been described previously. Since the mRNA levels do not change during the same period that the methionine synthase levels increase, regulation of this protein by its cofactor must be exerted posttranscriptionally. PMID- 10362527 TI - Effects of promoter mutations on the in vivo regulation of the cop operon of Enterococcus hirae by copper(I) and copper(II). AB - The cop operon of Enterococcus hirae encodes a repressor, CopY, a copper chaperone, CopZ, and two copper ATPases, CopA and CopB. Regulation of the cop operon is bi-phasic, with copper addition as well as copper chelation leading to induction. Using a plasmid-borne system with a reporter gene, induction of wild type and mutant cop promoters by high and low copper conditions was investigated. Only mutations that impaired the interaction of CopY with both DNA binding sites had a marked effect on regulation, leading to hyperinduction by copper(I) or copper(II). Chelation of copper(II), but not copper(I), also induced the operon, but induction by copper chelation was not significantly affected by the mutations. E. hirae mutants with reduced extracellular copper reductase activity exhibited the same induction kinetics as wild-type cells. These results show that copper addition and copper chelation induce the cop operon by different routes. PMID- 10362528 TI - Reactivation and refolding of a partially folded creatine kinase modified by 5,5' dithio-bis(2-nitrobenzoic acid). AB - Creatine kinase with its thiol groups modified by 5, 5'-dithio-bis(2-nitrobenzoic acid) has been shown to be partially folded in a monomeric state using fluorescence, circular dichroism, proteolysis, and size exclusion chromatography studies. In the presence of DTT, the partially folded modified creatine kinase can be reactivated and refolded following a biphasic course, suggesting the existence of a monomeric intermediate during the refolding of CK. The results provide evidence for our previously suggested model of the refolding pathway of urea-denatured creatine kinase. PMID- 10362529 TI - Tentative novel mechanism of the bystander effect in glioma gene therapy with HSV TK/GCV system. AB - Although many works support gap junctional intercellular communication (GJIC) having a close relation to bystander cell killing in herpes simplex virus thymidine kinase (HSV-TK) gene and ganciclovir (GCV) treatment, our previous work suggested that other factors involved in bystander effect besides GJIC exist. To confirm our primary work, we evaluated the mode of the bystander cell (C6) co cultured with TK-positive cells (TF10.2) in our designed "insert plates" in which two cell lines could be separated but share the same medium. Another method that we used was adding the supernatant from the medium of GCV-treated TF10.2 cells to the wild type C6. Growth inhibition of the bystander cells was observed despite the absence of GJIC. In addition, apoptotic cell death of TK+ cells and bystander cells was obvious. These studies suggested that other pathways besides cell-cell contacts may play a role in bystander cell killing; the factors released from TK positive cells could induce apoptosis of bystander cells. PMID- 10362530 TI - Peroxynitrite reacts with biological nitrogen-containing cyclic molecules by a radical pathway, as demonstrated by ultraweak luminescence coupled with ESR technique. AB - Ultraweak luminescence (uwCL) was coupled with electron spin resonance to study the reactions of 3 heterocyclic compounds (tryptophan, serotonin and imidazole) with peroxynitrite at pH 8.7. Tryptophan and serotonin reacted with emission of a flash peak of light (5 s) followed by a long-living light emission of +/- 80 s. Addition of the spin trap 4-POBN at different intervals, after the beginning of reaction revealed that a short-living free radical was produced in the case of serotonin and imidazole, but that with tryptophan, the initial radical rearranged into a relatively long-living radical, which was still formed when 4-POBN was added after 55 s (decreasing phase of uwCL). PMID- 10362531 TI - Activation of the neutrophil NADPH oxidase is inhibited by SB 203580, a specific inhibitor of SAPK2/p38. AB - Activation of the neutrophil NADPH oxidase by either the bacterial peptide fMLP or phorbol myristate acetate (PMA) is partially suppressed by SB 203580, a specific inhibitor of the MAP kinase family member, SAPK2/p38. The concentration of SB 203580 that suppresses activation of NADPH oxidase is similar to that which inhibits SAPK2/p38 in vitro, and both fMLP and PMA induce an extremely rapid and potent activation of SAPK2/p38 in neutrophils. SB 203580 does not exert its effect by preventing the neutrophil priming reaction, by suppressing the phosphorylation of p47phax, or by preventing the translocation of p47phax/p67phax to the plasma membrane. PMID- 10362532 TI - Functional domains of template-activating factor-I as a protein phosphatase 2A inhibitor. AB - Template-Activating Factor-I (TAF-I) alpha and beta, chromatin remodeling factors, were identified as the stimulatory factor for replication of the adenovirus DNA complexed with viral basic core proteins. Recently, two cellular inhibitors for protein phosphatase 2A (PP2A) have been isolated. One of these inhibitors, designated IPP2A2, is a truncated version of TAF-Ibeta. Here, it is shown using recombinant TAF-I proteins that both TAF-Ialpha and beta have the PP2A inhibitor activity. The N-terminal region but not the C-terminal acidic region, the latter of which is essential for the chromatin remodeling activity, is shown to be required for the PP2A inhibitor activity. Roles of TAF-Ialpha- and beta-specific regions, the C-terminal acidic region, and other regions of TAF-I for the PP2A inhibitor activity are also discussed. PMID- 10362533 TI - Regulation by interleukin-1beta of gene expression of bradykinin B1 receptor in MH-S murine alveolar macrophage cell line. AB - The effects of recombinant murine interleukin (IL)-1beta on gene expression of murine bradykinin B1 receptor (BDKRB1) in MH-S murine alveolar macrophage cell line were evaluated. BDKRB1 mRNA expression in MH-S cells was increased by IL 1beta (1, 3, and 10 ng/ml) in a time-dependent manner, peaking at 3-4 h by 100 1000 fold. IL-1beta (5 ng/ml, 24h) also induced significant binding to [3H]-des Arg10-kallidin with a dissociation constant (Kd) of 2.95 nM and a maximal binding density (Bmax) of 670 sites/cell. Des-Arg10-kallidin (10 microM), a BDKRB1 agonist, increased intracellular calcium ion ([Ca2+]i) in IL-1beta (5 ng/ml, 24 h)-exposed cells, an increase not observed in the cells not exposed to IL-1beta. A significant increase of tumor necrosis factor (TNF)-alpha secretion occurred in the IL-1beta (5 ng/ml, 24 h)-exposed cells following addition of des-Arg10 kallidin (the IL-1beta-exposed group: 57. 8 +/- 13.7 vs. the vehicle-exposed group: 16.7 +/- 4.3 pg/ml, p < 0.05 after a 100 nM des-Arg10-kallidin for 8 h), with an optimal effect at 3-100 nM. These data suggest that IL-1beta may up regulate BDKRB1-mediated functions of alveolar macrophages via an induction of BDKRB1 gene expression. PMID- 10362534 TI - Escherichia coli cafA gene encodes a novel RNase, designated as RNase G, involved in processing of the 5' end of 16S rRNA. AB - We found that the Escherichia coli cafA::cat mutant accumulated a precursor of 16S rRNA. This precursor migrated to the same position with 16.3S precursor found in the BUMMER strain that is known to be deficient in the 5' end processing of 16S rRNA. Accumulation of 16. 3S rRNA in the BUMMER mutant was complemented by introduction of a plasmid carrying the cafA gene. The mutant type cafA gene cloned from the BUMMER strain had a 11-bp deletion in its coding region. A small amount of the mature 16S rRNA was still formed in the cafA::cat mutant. This residual activity was found to be due to RNase E encoded by the rne/ams gene by rifampicin-chase experiments of the cafA::cat ams1 double mutant. These results indicated that the cafA gene encodes a novel RNase responsible for processing of the 5' end of 16S rRNA. PMID- 10362535 TI - Human estrogenic 17beta-hydroxysteroid dehydrogenase: predominance of estrone reduction and its induction by NADPH. AB - Human estrogenic 17beta-hydroxysteroid dehydrogenase (17beta-HSD1) plays a crucial role in the last step of the synthesis of estrogens. A detailed kinetic study demonstrated that the enzyme shows about 240 fold higher specificity towards estrone reduction than estradiol oxidation at physiological pH using tri phosphate cofactors. The kcat/Km values are 96 +/- 10 and 0.4 +/- 0.1 s-1 (microM)-1 respectively for the above two reactions. However, it has been shown that this difference is closely linked to the use of NADPH and NADP cofactors. A binding study using equilibrium dialysis indicated similar KD (equilibrium dissociation constant) of 11 +/- 1 and 4.7 +/- 0.9 microM for estrone and estradiol, respectively. The binding affinity of 17beta-HSD1 to estrone was significantly increased with a KD of 1.6 +/- 0.2 microM in the presence of NADP, the latter used as an analogue of the NADPH. The results of binding studies agree with the steady-state kinetics, which showed that the Km of estrone is 12-fold lower when using NADPH as a cofactor than when using NADH. These results strongly suggest that the cofactor plays a crucial role in the stimulation of the specificity for estrogen reduction. PMID- 10362536 TI - A functional role for ezrin during Shigella flexneri entry into epithelial cells. AB - Shigella flexneri is an enteroinvasive bacterium responsible for bacillary dysentery in humans. Bacterial entry into epithelial cells is a crucial step for the establishment of the infection. It is characterized by a transient reorganization of the host cell cytoskeleton at the site of bacterial interaction with the cell membrane, which leads to bacterial engulfment by a macropinocytic process. We show in this study that the membrane-cytoskeleton linker, ezrin, a member of the ERM (ezrin, radixin, moesin) family, plays an active role in the process of Shigella uptake. Ezrin is highly enriched in cellular protrusions induced by the bacterium and is found in close association with the plasma membrane. In addition, Shigella entry is significantly reduced in cells transfected with a dominant negative allele of ezrin with entry foci showing much shorter cellular protrusions. These results indicate that ezrin not only acts as a membrane-cytoskeleton linker, but may also mediate extension of cellular projections in the presence of signals such as those elicited by invading microorganisms. PMID- 10362537 TI - Rho family GTPases control entry of Shigella flexneri into epithelial cells but not intracellular motility. AB - Shigella flexneri, an invasive bacterial pathogen, promotes formation of two cytoskeletal structures: the entry focus that mediates bacterial uptake into epithelial cells and the actin-comet tail that enables the bacteria to spread intracellularly. During the entry step, secretion of bacterial invasins causes a massive burst of subcortical actin polymerization leading the formation of localised membrane projections. Fusion of these membrane ruffles leads to bacterial internalization. Inside the cytoplasm, polar expression of the IcsA protein on the bacterial surface allows polymerization of actin filaments and their organization into an actin-comet tail leading to bacterial spread. The Rho family of small GTPases plays an essential role in the organization and regulation of cellular cytoskeletal structures (i.e. filopodia, lamellipodia, adherence plaques and intercellular junctions). We show here that induction of Shigella entry foci is controlled by the Cdc42, Rac and Rho GTPases, but not by RhoG. In contrast, actin-driven intracellular motility of Shigella does not require Rho GTPases. Therefore, Shigella appears to manipulate the epithelial cell cytoskeleton both by Rho GTPase-dependent and -independent processes. PMID- 10362538 TI - Pervanadate-mediated tyrosine phosphorylation of keratins 8 and 19 via a p38 mitogen-activated protein kinase-dependent pathway. AB - Glandular epithelia express the keratin intermediate filament (IF) polypeptides 8, 18 and 19 (K8/18/19). These proteins undergo significant serine phosphorylation upon stimulation with growth factors and during mitosis, with subsequent modulation of their organization and interaction with associated proteins. Here we demonstrate reversible and dynamic tyrosine phosphorylation of K8 and K19, but not K18, upon exposure of intact mouse colon or cultured human cells to pervanadate. K8/19 tyrosine phosphorylation was confirmed by metabolic 32PO4-labeling followed by phosphoamino acid analysis, and by immunoblotting with anti-phosphotyrosine antibodies. Pervanadate treatment increases keratin solubility and also indirectly increases K8/18 serine phosphorylation at several known sites, some of which were previously shown to be associated with EGF stimulation, extracellular signal-regulated kinase (ERK), or p38 kinase activation. However, K8/19 tyrosine phosphorylation is independent of EGF signaling or ERK activation while inhibition of p38 kinase activity blocks pervanadate-induced K8/19 tyrosine phosphorylation. Our results demonstrate tyrosine phosphatase inhibitor-mediated in vivo tyrosine phosphorylation of K8/19, but not K18, and suggest that tyrosine phosphorylation may be a general modification of other IF proteins. K8/19 tyrosine phosphorylation involves a pathway that utilizes the p38 mitogen-activated protein kinase, but appears independent of EGF signaling or ERK kinase activation. PMID- 10362539 TI - Processing of CFTR bearing the P574H mutation differs from wild-type and deltaF508-CFTR. AB - Cystic fibrosis transmembrane conductance regulator (CFTR) containing the deltaF508 mutation is retained in the endoplasmic reticulum (ER). This defect can be partially overcome by a reduction in temperature which allows some of the deltaF508 protein to exit the ER and move to the cell surface. Earlier studies showed that the CF-associated mutants, P574H and A455E, were also misprocessed. In this study, we found that processing of P574H and A455E was also temperature sensitive; at 26 degrees C, some of the protein matured. In contrast to other CFTR mutants, P574H accumulated in punctate cytoplasmic bodies that colocalized with endoplasmic reticulum (ER) markers. At 26 degrees C, these bodies were no longer present. P574H showed a prolonged association with Hsp70 and also colocalized with Hsp70. We used brefeldin A (BFA) to determine which processing step(s) was altered by reduced temperature. Unlike wild-type CFTR, which was converted into an intermediate that was stable in the presence of BFA at 37 degrees C, deltaF508 and P574H produced the intermediate only when the temperature was reduced to 26 degrees C. Furthermore the wild-type intermediate was not associated with Hsp70. These data suggest that formation of the stable intermediate is a key temperature-sensitive step and appears to be coincident with release of the wild-type protein from Hsp70. PMID- 10362540 TI - Intermediate filament interactions can be altered by HSP27 and alphaB-crystallin. AB - HSP27 and alphaB-crystallin are both members of the small heat shock protein family. alphaB-crystalllin has been proposed to modulate intermediate filaments and recently a mutation in alphaB-crystallin has been identified as the genetic basis of desmin related myopathy. This disease is characterised in its pathology by aggregates of intermediate filaments associated with alphaB-crystallin. Here we report that HSP27 like alphaB-crystallin is associated with glial fibrillary acidic protein and vimentin intermediate filament networks in unstressed U373MG astrocytoma cells. HSP27 is also associated with keratin filaments in MCF7 cells, indicating that this association is not restricted to a particular intermediate filament type. The association of sHSPs with both the soluble and filamentous intermediate filament fractions of U373 cells was demonstrated biochemically. Heat shock or drug treatments induced a co-collapse of intermediate filaments and associated small heat shock proteins. These data show that the presence of HSP27 or alphaB-crystallin could not prevent filament collapse and suggest that the purpose of this association is more than just filament binding. Indeed, in U373MG cells the intermediate filament association with small heat shock proteins is similar to that observed for another protein chaperone, HSC70. In order to discern the effect of different chaperone classes on intermediate filament network formation and maintenance, several in vitro assays were assessed. Of these, falling ball viscometry revealed a specific activity of small heat shock proteins compared to HSC70 that was apparently inactive in this assay. Intermediate filaments form a gel in the absence of small heat shock proteins. In contrast, inclusion of alphaB-crystallin or HSP27 prevented gel formation but not filament assembly. The transient transfection of GFAP into MCF7 cells was used to show that the induction of a completely separate network of intermediate filaments resulted in the specific association of the endogenous HSP27 with these new GFAP filaments. These data lead us to propose that one of the major functions of the association of small heat shock proteins with intermediate filaments is to help manage the interactions that occur between filaments in their cellular networks. This is achieved by protecting filaments against those non-covalent interactions that result when they come into very close proximity as seen from the viscosity experiments and which have the potential to induce intermediate filament aggregation as seen in some disease pathologies. PMID- 10362541 TI - Insect cuticle, an in vivo model of protein trafficking. AB - In the course of this study more than 20 proteins have been isolated from the larval cuticle of Manduca sexta. Synthesis, secretion, transport and accumulation of four particular proteins, representative members of four characteristic groups, were followed during metamorphosis by immunoblot and immuncytochemical methods and are described in detail in this paper. We established that only some of the proteins of the soft cuticle of Lepidopteran larvae are synthesized in epidermal cells at the beginning of the larval stages and are digested during the moulting period (MsCP29). Other proteins (MsCP30/11) are secreted into the cuticle by the epidermal cells in different forms during various developmental stages. Some proteins are secreted apically during the feeding period, but before ecdysis they are then taken up by epidermal cells and transported in a basolateral direction back into the hemolymph and saved in an immunologically intact form by the fat body cells (MsCP12.3). Some cuticle proteins have a non epidermal origin. They are transported from the hemolymph into the cuticle. Before and during ecdysis these molecules reappear in the hemolymph and are detectable again in the pupal cuticle (MsCP78). Our data prove that the cuticle is not a non-living part of the insect body: it is not only an inert, protective armor, but maintains a continuous and dynamic metabolic connection with the other organs of the organism. PMID- 10362542 TI - Domain analysis of supervillin, an F-actin bundling plasma membrane protein with functional nuclear localization signals. AB - A growing number of actin-associated membrane proteins have been implicated in motile processes, adhesive interactions, and signal transduction to the cell nucleus. We report here that supervillin, an F-actin binding protein originally isolated from bovine neutrophil plasma membranes, contains functional nuclear targeting signals and localizes at or near vinculin-containing focal adhesion plaques in COS7-2 and CV1 cells. Overexpression of full-length supervillin in these cells disrupts the integrity of focal adhesion plaques and results in increased levels of F-actin and vinculin. Localization studies of chimeric proteins containing supervillin sequences fused with the enhanced green fluorescent protein indicate that: (1) the amino terminus promotes F-actin binding, targeting to focal adhesions, and limited nuclear localization; (2) the dominant nuclear targeting signal is in the center of the protein; and (3) the carboxy-terminal villin/gelsolin homology domain of supervillin does not, by itself, bind tightly to the actin cytoskeleton in vivo. Overexpression of chimeras containing both the amino-terminal F-actin binding site(s) and the dominant nuclear targeting signal results in the formation of large nuclear bundles containing F-actin, supervillin, and lamin. These results suggest that supervillin may contribute to cytoarchitecture in the nucleus, as well as at the plasma membrane. PMID- 10362543 TI - Localisation of presenilin 2 in human and rodent pancreatic islet beta-cells; Met239Val presenilin 2 variant is not associated with diabetes in man. AB - Mutations in presenilin 1 and 2 are causative factors for early onset familial Alzheimer's disease and possible roles for presenilins include protein trafficking, regulation of apoptosis and/or calcium homeostasis. Presenilin 2 mRNA is expressed in brain, muscle and pancreas but the role of pancreatic presenilin 2 and its relationship to diabetes are unknown. Presenilin 2 immunoreactivity was localised in human and rodent pancreas to islet cells and found in granules of beta-cells. Presenilin 2 was identified in primitive islet and duct cells of human foetal pancreas and in proliferating exocrine duct cells in human pancreatitis but not found in islet amyloid deposits in Type 2 diabetic subjects. Full length, approximately 50 kDa, and the approximately 30 kDa N terminal fragment of presenilin 2 were identified by western blotting in extracted rodent pancreas but only the 30 kDa fragment was detected in mouse islets and human insulinoma. Post-mortem pancreatic morphology was normal in a subject with the presenilin 2 Met239Val variant and early onset familial Alzheimer's disease. Oral glucose tolerance tests on subjects with the presenilin 2 Met239Val mutation unaffected by early onset familial Alzheimer's disease (mean age 35 years) and on their first-degree relatives without the mutation demonstrated no evidence of glucose intolerance or increased proinsulin secretion. PS2 is a novel &bgr;-cell protein with potential roles in development or protein processing but pancreatic islet structure and function appear to be unaffected by the Met239Val mutation. PMID- 10362544 TI - 3-D organization of ribosomal transcription units after DRB inhibition of RNA polymerase II transcription. AB - In each bead of the nucleolar necklace, using adenosine analog DRB-treated PtK1 cells, we investigated the three components of rDNA transcription, i.e. the gene, transcription factor UBF and transcripts. In situ hybridization revealed the unraveling and 3-D dispersion of most of the rDNA coding sequences within the nucleus. The signals were small, of similar intensity and tandemly organized in the necklace. This observation is compatible with the fact that they might correspond to single gene units. Active transcription was visualized in these units, demonstrating that they were active functional units. Transcript labeling was not similar for each unit, contrary to UBF labeling. UBF and rRNA transcripts were only partially colocalized, as demonstrated by 3-D image analysis and quantification. As visualized by electron microscopy, the necklace was composed of a small fibrillar center partially surrounded by a dense fibrillar component. The 3-D arrangement of this individual unit in the necklace, investigated both by confocal and electron microscopy in the same cells, showed that the individual beads were linked by a dense fibrillar component. The reversibility of this organization after removal of DRB indicated that the beads in the necklace are certainly the elementary functional domain of the nucleolus. In addition, these results lead us to suggest that the organization of a functional domain, presumably corresponding to a single gene, can be studied by in situ approaches. PMID- 10362545 TI - Molecular architecture of the lens fiber cell basal membrane complex. AB - Lens fiber cells are transparent, highly elongated, epithelial cells. Because of their unusual length these cells represent a novel model system to investigate aspects of epithelial cell polarity. In this study, we examined the fiber cell basal membrane complex (BMC). The BMC anchors fiber cells to the lens capsule and facilitates their migration across the capsule. Confocal microscopy revealed that bundled actin filaments converge beneath the center of each BMC and insert into the lateral membrane at points enriched in N-cadherin. Two other contractile proteins, caldesmon and myosin, were enriched in the BMC, co-localizing with f actin bundles. The actin/N-cadherin complex formed a hexagonal lattice, cradling the posterior face of the lens. Removal of the capsule caused the tips of the fiber cells to break off, remaining attached to the stripped capsule. This provided a method for assaying cell adhesion and purifying BMC components. Fiber cell adhesion required Mg2+ and/or Ca2+ and was disrupted by incubation with beta1 integrin antibody. BMC proteins were compared with samples from the neighboring lateral membrane. Although some components were common to both samples, others were unique to the BMC. Furthermore, some lateral membrane proteins, most notably lens major intrinsic protein (MIP), were excluded from the BMC. Western blotting of BMC preparations identified several structural proteins originally found in focal adhesions and two kinases, FAK and MLCK, previously undescribed in the lens. These data suggest that the BMC constitutes a distinct membrane domain in the lens. The structural organization of the BMC suggests a role in shaping the posterior lens face and hence the refractive properties of the eye. PMID- 10362546 TI - Receptor mediated and fluid phase pathways for internalization of the ER Hsp90 chaperone GRP94 in murine macrophages. AB - Immunization of mice with GRP94, the endoplasmic reticulum (ER) Hsp90, elicits cytotoxic T lymphocyte (CTL) responses to chaperone-bound, source cell-derived peptides. Elicitation of a CTL response requires that GRP94-associated peptides be transferred onto major histocompatability complex (MHC) class I molecules, a process that is postulated to accompany GRP94 internalization by antigen presenting cells, such as macrophages (Mphi) and dendritic cells (DC). In studies of GRP94 uptake in elicited Mphi, we report that Mphi display specific cell surface binding of GRP94, and that surface-bound GRP94 can be internalized via receptor mediated endocytosis. GRP94 internalized by this pathway co-localized predominately with transferrin-positive early endosomes. At time periods of up to 20 minutes, little trafficking of GRP94 to the lysosomal compartment was observed. When GRP94 was present in the medium, and thus accessible to both receptor-mediated and fluid phase internalization pathways, internalization was modestly inhibited in the presence of yeast mannan, a competitive inhibitor of mannose/fucose receptor activity, and substantially inhibited by dimethylamiloride, an inhibitor of macropinocytosis. GRP94 internalized via macropinocytosis did not display prominent co-staining with the lysosomal marker LAMP-2. These data identify multiple pathways of GRP94 internalization and indicate that receptor-dependent uptake of GRP94 is not dependent upon its high mannose oligosaccharide moiety. Most significantly, these data demonstrate the existence of cell surface receptor(s), apparently unique to antigen presenting cells, that function in the binding and internalization of the ER chaperone GRP94. PMID- 10362547 TI - RHO-associated protein kinase alpha potentiates insulin-induced MAP kinase activation in Xenopus oocytes. AB - We recently identified Xenopus Rho-associated protein kinase alpha (xROKalpha) as a Xenopus insulin receptor substrate-1 binding protein and demonstrated that the non-catalytic carboxyl terminus of xROKalpha binds Xenopus insulin receptor substrate-1 and blocks insulin-induced MAP kinase activation and germinal vesicle breakdown in Xenopus oocytes. In the current study we further examined the role of xROKalpha in insulin signal transduction in Xenopus oocytes. We demonstrate that injection of mRNA encoding the xROKalpha kinase domain or full length xROKalpha enhanced insulin-induced MAP kinase activation and germinal vesicle breakdown. In contrast, injection of a kinase-dead mutant of xROKalpha or pre incubation of oocytes with an xROKalpha inhibitor significantly reduced insulin induced MAP kinase activation. To further dissect the mechanism by which xROKalpha may participate in insulin signalling, we explored a potential function of xROKalpha in regulating cellular Ras function, since insulin-induced MAP kinase activation and germinal vesicle breakdown is known to be a Ras-dependent process. We demonstrate that whereas injection of mRNA encoding c-H-Ras alone induced xMAP kinase activation and GVBD in a very low percentage (about 10%) of injected oocytes, co-injection of mRNA encoding xROKalpha and c-H-Ras induced xMAP kinase activation and germinal vesicle breakdown in a significantly higher percentage (50-60%) of injected oocytes. These results suggest a novel function for xROKalpha in insulin signal transduction upstream of cellular Ras function. PMID- 10362548 TI - Isolation of tubulin polyglutamylase from Crithidia; binding to microtubules and tubulin, and glutamylation of mammalian brain alpha- and beta-tubulins. AB - Trypanosomatids have a striking cage-like arrangement of submembraneous microtubules. We previously showed that alpha- and beta- tubulins of these stable microtubules are extensively modified by polyglutamylation. Cytoskeletal microtubular preparations obtained by Triton extraction of Leishmania tarentolae and Crithidia fasciculata retain an enzymatic activity that incorporates radioactive glutamic acid in a Mg2+-ATP-dependent manner into alpha- and beta tubulins. The tubulin polyglutamylase is extracted by 0.25 M salt. The Crithidia enzyme can be purified by ATP-affinity chromatography, glycerol-gradient centrifugation and ion-exchange chromatography. After extraction from the microtubular cytoskeleton the glutamylase forms a complex with alphabeta tubulin, but behaves after removal of tubulin as a globular protein with a molecular mass of 38x10(3). In highly enriched fractions a corresponding band is the major polypeptide visible in SDS-PAGE. The enzyme from Crithidia recognises mammalian brain tubulin, where it incorporates glutamic acid preferentially into the more acidic variants of both alpha- and beta-tubulins. Synthetic peptides with an oligoglutamyl side chain, corresponding to the carboxy-terminal end of brain alpha- and beta-tubulins, are accepted by the enzyme, albeit at low efficiency. The polyglutamylase elongates the side chain by up to 3 and 5 residues, respectively. Other properties of the tubulin polyglutamylase are also discussed. PMID- 10362549 TI - Role of myosin II tail sequences in its function and localization at the cleavage furrow in Dictyostelium. AB - Cytoplasmic myosin II accumulates in the cleavage furrow and provides the force for cytokinesis in animal and amoeboid cells. One model proposes that a specific domain in the myosin II tail is responsible for its localization, possibly by interacting with a factor concentrated in the equatorial region. To test this possibility, we have expressed myosins carrying mutations in the tail domain in a strain of Dictyostelium cells from which the endogenous myosin heavy chain gene has been deleted. The mutations used in this study include four internal tail deletions: Mydelta824-941, Mydelta943-1464, Mydelta943-1194 and Mydelta1156-1464. Contrary to the prediction of the hypothesis, immunofluorescence staining demonstrated that all mutant myosins were able to move toward the furrow region. Chimeric myosins, which consisted of a Dictyostelium myosin head and chicken skeletal myosin tail, also efficiently localized to the cleavage furrow. All these deletion and chimeric mutant myosins, except for Mydelta943-1464, the largest deletion mutant, were able to support cytokinesis in suspension. Our data suggest that there is no single specific domain in the tail of Dictyostelium myosin II that is required for its functioning at and localization to the cleavage furrow. PMID- 10362550 TI - The Drosophila forked protein induces the formation of actin fiber bundles in vertebrate cells. AB - The forked protein is an actin binding protein involved in the formation of large actin fiber bundles in developing Drosophila bristles. These are the largest example of a type of actin bundle characterized by parallel, hexagonally packed actin fibers, also found in intestinal microvilli, kidney proximal tubule microvilli, and stereocilia in the ear. Understanding how these structures are constructed and how that construction is regulated is an important question in cell and developmental biology. Because the timing of forked gene expression coincides with the formation of the actin fiber bundles, and since the forked protein is localized at the site of initiation of these bundles before they form, it has been proposed that the forked protein is an initiator of actin bundle formation. In this paper we show that the forked protein can induce the formation of bundles and increase actin polymerization in vertebrate cells. We use this system to identify regions of the forked protein which are essential for bundle formation and actin co-localization. PMID- 10362551 TI - Overexpression of class I, II or IVb beta-tubulin isotypes in CHO cells is insufficient to confer resistance to paclitaxel. AB - Recent studies have suggested a correlation between increased expression of specific beta-tubulin isotypes and paclitaxel resistance in drug-selected cell lines. In an attempt to establish a causal link, we have transfected Chinese hamster ovary cells with cDNAs encoding epitope-tagged class I, II, and IVb beta tubulins, as well as a class I beta-tubulin with a mutation previously characterized in a paclitaxel resistant mutant. To eliminate possible toxicity that might be associated with overexpression of non-native tubulin, each of the cDNAs was placed under the control of a tetracycline regulated promoter. All transfected cDNAs produced assembly competent tubulin whose synthesis could be turned off or on by the presence or absence of tetracycline. Production of betaI, betaII, or betaIVb tubulin had no effect on the sensitivity of the cells to paclitaxel, but production of the mutant betaI-tubulin conferred clear resistance to the drug. We conclude from these experiments that simple overexpression of class I, II, or IVb isoforms of beta-tubulin is insufficient to confer resistance to paclitaxel. PMID- 10362552 TI - Induction of apoptosis by overexpression of the DNA-binding and DNA-PK-activating protein C1D. AB - Apoptosis is induced in various tumor cell lines by vector-dependent overexpression of the conserved gene C1D that encodes a DNA-binding and DNA-PK activating protein. C1D is physiologically expressed in 50 human tissues tested, which points to its basic cellular function. The expression of this gene must be tightly regulated because elevated levels of C1D protein, e.g. those induced by transient vector-dependent expression, result in apoptotic cell death. Cells transfected with C1D-expressing constructs show terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling of DNA ends. Transfections with constructs in which C1D is expressed in fusion with the (enhanced) green fluorescent protein from A. victoria (EGFP) allow the transfected cells to be identified and the morphological changes induced to be traced. Starting from intense nuclear spots, green fluorescence reflecting C1D expression increases dramatically at 12-24 hours post-transfection. Expression of C1D-EGFP protein is accompanied by morphological changes typical of apoptotic cell death, e.g. cytoplasmic vacuolation, membrane blebbing and nuclear disintegration. Cell shrinkage and detachment from extracellular matrix are observed in monolayer cultures while suspension cells become progressively flattened. The facility to differentiate between transfected and non-transfected cells reveals that non transfected cells co-cultured with transfected cells also show the morphological changes of apoptosis, which points to a bystander effect. C1D-dependent apoptosis is not induced in cells with non-functional p53. Accordingly, C1D-induced apoptosis is discussed in relation to its potential to activate DNA-PK, which has been considered to act as an upstream activator of p53. PMID- 10362553 TI - Analysis of the roles of the head domains of type IV rat neuronal intermediate filament proteins in filament assembly using domain-swapped chimeric proteins. AB - Type IV neuronal intermediate filament proteins consist of alpha-internexin, which can self-assemble into filaments and the neurofilament triplet proteins, which are obligate heteropolymers, at least in rodents. These IF proteins therefore provide good systems for elucidating the mechanism of intermediate filament assembly. To analyze the roles of the head domains of these proteins in contributing to their differential assembly properties, we generated chimeric proteins by swapping the head domains between rat alpha-internexin and either rat NF-L or NF-M and examined their assembly properties in transfected cells that lack their own cytoplasmic intermediate filament network. Lalphaalpha and Malphaalpha, the chimeric proteins generated by replacing the head domain of alpha-internexin with those of NF-L and NF-M, respectively, were unable to self assemble into filaments. In contrast, alphaLL, a chimeric NF-L protein generated by replacing the head domain of NF-L with that of alpha-internexin, was able to self-assemble into filaments, whereas MLL, a chimeric NF-L protein containing the NF-M head domain, was unable to do so. These results demonstrate that the alpha internexin head domain is essential for alpha-internexin's ability to self assemble. While coassembly of Lalphaalpha with NF-M and coassembly of Malphaalpha with NF-L resulted in formation of filaments, coassembly of Lalphaalpha with NF-L and coassembly of Malphaalpha with NF-M yielded punctate patterns. These coassembly results show that heteropolymeric filament formation requires that one partner has the NF-L head domain and the other partner has the NF-M head domain. Thus, the head domains of rat NF-L and NF-M play important roles in determining the obligate heteropolymeric nature of filament formation. The data obtained from these self-assembly and coassembly studies provide some new insights into the mechanism of intermediate filament assembly. PMID- 10362554 TI - Hyaluronan stimulates tumor cell migration by modulating the fibrin fiber architecture. AB - The glycosaminoglycan hyaluronan, which supports tumor cell migration and metastasis, interferes with fibrin polymerization and leads to increased fiber size and porosity of fibrin clots. Here we have studied the proportionate effect of fibrin polymerization on hyaluronan-mediated migration of glioblastoma cells. The structural and physical properties of hyaluronan-containing fibrin gels were analyzed by turbidity measurement, laser scanning microscopy, compaction assay, and calculation of pore size by liquid permeation. When fibrin polymerized in the presence of hyaluronan or dextran, the resulting gels strongly stimulated cell migration, and migration significantly correlated with fiber mass-to-length ratios and pore diameters. In contrast, cell migration was not induced by addition of hyaluronan to supernatants of already polymerized gels. Hyaluronan mediated migration was inhibited in fibrin gels by antibodies to alphav- and beta1integrins and the disintegrin echistatin, but not by antibodies to the hyaluronan receptor CD44 (up to 50 microg/ml). As a control, we show that anti CD44 (10 microg/ml) inhibited cell migration on a pure hyaluronan matrix using a two-dimensional Boyden chamber system. In contrast to three-dimensional migration, the migration of cells on the surfaces of variably structured fibrin gels was not significantly different, indicating that increased gel permeability (porosity) may account for hyaluronan-mediated migration. We conclude that, in complex three-dimensional substrates, the predominant effect of hyaluronan on cell migration might be indirect and requires modulation of fibrin polymerization. PMID- 10362555 TI - Sequential recruitment of NPC proteins to the nuclear periphery at the end of mitosis. AB - Nuclear pore complexes (NPCs) are extremely elaborate structures that mediate the bidirectional movement of macromolecules between the nucleus and cytoplasm. With a mass of about 125 MDa, NPCs are thought to be composed of 50 or more distinct protein subunits, each present in multiple copies. During mitosis in higher cells the nuclear envelope is disassembled and its components, including NPC subunits, are dispersed throughout the mitotic cytoplasm. At the end of mitosis, all of these components are reutilized. Using both conventional and digital confocal immunofluorescence microscopy we have been able to define a time course of post mitotic assembly for a group of NPC components (CAN/Nup214, Nup153, POM121, p62 and Tpr) relative to the integral nuclear membrane protein LAP2 and the NPC membrane glycoprotein gp210. Nup153, a component of the nuclear basket, associates with chromatin towards the end of anaphase, in parallel with the inner nuclear membrane protein, LAP2. However, immunogold labeling suggests that the initial Nup153 chromatin association is membrane-independent. Assembly of the remaining proteins follows that of the nuclear membranes and occurs in the sequence POM121, p62, CAN/Nup214 and gp210/Tpr. Since p62 remains as a complex with three other NPC proteins (p58, 54, 45) during mitosis and CAN/Nup214 maintains a similar interaction with its partner, Nup84, the relative timing of assembly of these additional four proteins may also be inferred. These observations suggest that there is a sequential association of NPC proteins with chromosomes during nuclear envelope reformation and the recruitment of at least eight of these precedes that of gp210. These findings support a model in which it is POM121 rather than gp210 that defines initial membrane-associated NPC assembly intermediates. PMID- 10362584 TI - Molecular mechanisms of cytoadherence in malaria. AB - Microbial pathogens subvert host adhesion molecules to disseminate or to enter host cells to promote their own survival. One such subversion is the cytoadherence of Plasmodium falciparum-infected erythrocytes (IRBC) to vascular endothelium, which protects the parasite from being removed by the spleen. The process results in microcirculatory obstruction and subsequent hypoxia, metabolic disturbances, and multiorgan failure, which are detrimental to the host. Understanding the molecular events involved in these adhesive interactions is therefore critical both in terms of pathogenesis and implications for therapeutic intervention. Under physiological flow conditions, cytoadherence occurs in a stepwise fashion through parasite ligands expressed on the surface of IRBC and the endothelial receptors CD36, intercellular adhesion molecule-1 (ICAM-1), P selectin, and vascular adhesion molecule-1. Moreover, rolling on ICAM-1 and P selectin increases subsequent adhesion to CD36, indicating that receptors can act synergistically. Cytoadherence may activate intracellular signaling pathways in both endothelial cells and IRBC, leading to gene expression of mediators such as cytokines, which could modify the outcome of the infection. PMID- 10362585 TI - Intestinal cell differentiation: cellular mechanisms and the search for the perfect model focus on "involvement of p21(WAF1/Cip1) and p27(Kip1) in intestinal epithelial cell differentiation". PMID- 10362586 TI - Involvement of p21(WAF1/Cip1) and p27(Kip1) in intestinal epithelial cell differentiation. AB - Using the conditionally immortalized human cell line tsFHI, we have investigated the role of cyclin-dependent kinase inhibitors (CKIs) in intestinal epithelial cell differentiation. Expression of cyclins, cyclin-dependent kinases (Cdk), and CKIs was examined under conditions promoting growth, growth arrest, or expression of differentiated traits. Formation of complexes among cell cycle regulatory proteins and their kinase activities were also investigated. The tsFHI cells express three CKIs: p16, p21, and p27. With differentiation, p21 and p27 were strongly induced, but with different kinetics: the p21 increase was rapid but transient and the p27 increase was delayed but sustained. Our results suggest that the function of p16 is primarily to inhibit cyclin D-associated kinases, making tsFHI cells dependent on cyclin E-Cdk2 for pRb phosphorylation and G1/S progression. Furthermore, they indicate that p21 is the main CKI involved in irreversible growth arrest during the early stages of cell differentiation in association with D-type cyclins, cyclin E, and Cdk2, whereas p27 may induce or stabilize expression of differentiated traits acting independently of cyclin-Cdk function. PMID- 10362587 TI - Ca2+-activated Cl- channels: focus on "molecular cloning and transmembrane structure of hCLCA2 from human lung, trachea, and mammary gland". PMID- 10362588 TI - Molecular cloning and transmembrane structure of hCLCA2 from human lung, trachea, and mammary gland. AB - The CLCA family of Ca2+-activated Cl- channels has recently been discovered, with an increasing number of closely related members isolated from different species. Here we report the cloning of the second human homolog, hCLCA2, from a human lung cDNA library. Northern blot and RT-PCR analyses revealed additional expression in trachea and mammary gland. A primary translation product of 120 kDa was cleaved into two cell surface-associated glycoproteins of 86 and 34 kDa in transfected HEK-293 cells. hCLCA2 is the first CLCA homolog for which the transmembrane structure has been systematically studied. Glycosylation site scanning and protease protection assays revealed five transmembrane domains with a large, cysteine-rich, amino-terminal extracellular domain. Whole cell patch-clamp recordings of hCLCA2-transfected HEK-293 cells detected a slightly outwardly rectifying anion conductance that was increased in the presence of the Ca2+ ionophore ionomycin and inhibited by DIDS, dithiothreitol, niflumic acid, and tamoxifen. Expression in human trachea and lung suggests that hCLCA2 may play a role in the complex pathogenesis of cystic fibrosis. PMID- 10362589 TI - Nitric oxide modulation of focal adhesions in endothelial cells. AB - A permissive role of nitric oxide (NO) in endothelial cell migration and angiogenesis promoted by vascular endothelial growth factor (VEGF), endothelin, and substance P has previously been established. The present studies were designed to examine the mechanism(s) involved in the NO effect on focal adhesions. Time-lapse videomicroscopy of human umbilical vein endothelial cells (HUVECs) plated on the silicone rubber substrate revealed that unstimulated cells were constantly remodeling the wrinkling pattern, indicative of changing tractional forces. Application of NO donors reversibly decreased the degree of wrinkling, consistent with the release of tractional forces exerted by focal adhesions and stress fibers. Morphometric and immunocytochemical analyses showed that NO inhibited adhesion and spreading of HUVECs and attenuated recruitment of paxillin to focal adhesions. NO also had a profound dose-dependent effect on the formation of stress fibers by HUVECs. De novo formation of focal adhesions in HUVECs was significantly diminished in the presence of NO donors. Migration of HUVECs showed an absolute requirement for the functional NO synthase. NO donors did not interfere with focal adhesion kinase recruitment to focal adhesions but affected the state of its tyrosine phosphorylation, as judged from the results of immunoprecipitation and immunoblotting experiments. Videomicroscopy of HUVECs presented with VEGF in a micropipette showed that the rate of cell migration was slowed down by NO synthase inhibition as well as by inhibition of tyrosine phosphorylation. Collectively, these data indicate that NO reversibly releases tractional forces exerted by spreading endothelial cells via interference with the de novo formation of focal adhesions, tyrosine phosphorylation of components of focal adhesion complexes, and assembly of stress fibers. PMID- 10362590 TI - Isoproterenol potentiates alpha-adrenergic and muscarinic receptor-mediated Ca2+ response in rat parotid cells. AB - The effects of the cAMP pathway on the Ca2+ response elicited by phospholipase C coupled receptor stimulations were studied in rat parotid cells. Although 1 microM isoproterenol (Iso) itself had no effect on the cytosolic Ca2+ concentration, the pretreatment with Iso potentiated Ca2+ responses evoked by phenylephrine. The potentiating effect of Iso was attributed to a shifting of the concentration-response curves of phenylephrine to the left and an increase in the maximal response. Half-maximal potentiation occurred at 3 nM Iso. Iso also potentiated the Ca2+ response elicited by carbachol. The potentiating effect of Iso was mimicked by forskolin (10 microM) and dibutyryl adenosine 3',5'-cyclic monophosphate (2 mM) and was blocked by 10 microM H-89. Iso potentiated the phenylephrine-induced Ca2+ response in the absence of extracellular Ca2+, but Iso did not increase the inositol trisphosphate (IP3) production induced by phenylephrine. These results suggest that the potentiation of the Ca2+ response can be attributed to a sensitization of IP3 receptors by cAMP-dependent protein kinase. PMID- 10362591 TI - Choline uptake across the ventricular membrane of neonate rat choroid plexus. AB - The uptake of [3H]choline from the cerebrospinal fluid (CSF) side of the rat neonatal choroid plexus was characterized in primary cultures of the choroidal epithelium grown on solid supports. Cell-to-medium concentration ratios were approximately 5 at 1 min and as high as 70 at 30 min. Apical choline uptake was facilitated; the Km was approximately 50 microM. Several organic cations (e.g., hemicholinium-3 and N1-methylnicotinamide) inhibited uptake. The reduction or removal of external Na+ or the addition of 5 mM LiCl had no effect on uptake. However, increasing external K+ concentration from 3 to 30 mM depolarized ventricular membrane potential (-70 to -15 mV) and reduced uptake to 45% of that for the control. Treatment with 1 mM ouabain or 2 mM BaCl2 reduced uptake 45%, and intracellular acidification reduced uptake to approximately 90% of that for controls. These data indicate that the uptake of choline from CSF across the ventricular membrane of the neonatal choroidal epithelium is not directly coupled to Na+ influx but is sensitive to plasma membrane electrical potential. PMID- 10362592 TI - Regulation of total mitochondrial Ca2+ in perfused liver is independent of the permeability transition pore. AB - Triggering of the permeability transition pore (PTP) in isolated mitochondria causes release of matrix Ca2+, ions, and metabolites, and it has been proposed that the PTP mediates mitochondrial Ca2+ release in intact cells. To study the role of the PTP in mitochondrial energy metabolism, the mitochondrial content of Ca2+, Mg2+, ATP, and ADP was determined in hormonally stimulated rat livers perfused with cyclosporin A (CsA). Stimulation of livers perfused in the absence of CsA with glucagon and phenylephrine induced an extensive uptake of Ca2+, Mg2+, and ATP plus ADP by the mitochondria, followed by a release on omission of hormones. In the presence of CsA, the PTP was fully inhibited, but neither the hormone-induced uptake of Ca2+, ATP, or ADP by mitochondria nor their release after washout of hormones was significantly changed. We conclude that the regulation of sustained changes in mitochondrial Ca2+ content induced by hormonal stimulation is independent of the PTP. PMID- 10362593 TI - ATP dependence is not an intrinsic property of Na+/H+ exchanger NHE1: requirement for an ancillary factor. AB - Na+/H+ exchange is a passive process not requiring expenditure of metabolic energy. Nevertheless, depletion of cellular ATP produces a marked inhibition of the antiport. No evidence has been found for direct binding of nucleotide to exchangers or alteration in their state of phosphorylation, suggesting ancillary factors may be involved. This possibility was tested by comparing the activity of dog red blood cells (RBC) and their resealed ghosts. Immunoblotting experiments using isoform-specific polyclonal and monoclonal antibodies indicated RBC membranes express Na+/H+ exchanger isoform 1 (NHE1). In intact RBC, uptake of Na+ was greatly stimulated when the cytosol was acidified. The stimulated uptake was largely eliminated by amiloride and by submicromolar concentrations of the benzoyl guanidinium compound HOE-694, consistent with mediation by NHE1. Although exchange activity could also be elicited by acidification in resealed ghosts containing ATP, the absolute rate of transport was markedly diminished at comparable pH. Dissipation of the pH gradient was ruled out as the cause of diminished transport rate in ghosts. This was accomplished by a "pH clamping" procedure based on continued export of base equivalents by the endogenous anion exchanger. These observations suggest a critical factor required to maintain optimal Na+/H+ exchange activity is lost or inactivated during preparation of ghosts. Depletion of ATP, achieved by incubation with 2-deoxy-D-glucose, inhibited Na+/H+ exchange in intact RBC, as reported for nucleated cells. In contrast, the rate of exchange was similar in control and ATP-depleted resealed ghosts. Interestingly, the residual rate of Na+/H+ exchange in ATP-depleted but otherwise intact cells was similar to the transport rate of ghosts. Therefore, we tentatively conclude that full activation of NHE1 requires both ATP and an additional regulatory factor, which may mediate the action of the nucleotide. Ancillary phosphoproteins or phospholipids or the kinases that mediate their phosphorylation are likely candidates for the regulatory factor(s) that is inactivated or missing in ghosts. PMID- 10362594 TI - Rho controls actin cytoskeletal assembly in renal epithelial cells during ATP depletion and recovery. AB - Actin cytoskeletal disruption is a hallmark of ischemic injury and ATP depletion in a number of cell types, including renal epithelial cells. We manipulated Rho GTPase signaling by transfection and microinjection in LLC-PK proximal tubule epithelial cells and observed actin cytoskeletal organization following ATP depletion or recovery by confocal microscopy and quantitative image analysis. ATP depletion resulted in disruption of stress fibers, cortical F-actin, and apical actin bundles. Constitutively active RhoV14 prevented disruption of stress fibers and cortical F-actin during ATP depletion and enhanced the rate of stress fiber reassembly during recovery. Conversely, the Rho inhibitor C3 or dominant negative RhoN19 prevented recovery of F-actin assemblies upon repletion. Actin bundles in the apical microvilli and cytosolic F-actin were not affected by Rho signaling. Assembly of vinculin and paxillin into focal adhesions was disrupted by ATP depletion, and constitutively active RhoV14, although protecting stress fibers from disassembly, did not prevent dispersion of vinculin and paxillin, resulting in uncoupling of stress fiber and focal adhesion assembly. We propose that ATP depletion causes Rho inactivation during ischemia and that recovery of normal cellular architecture and function requires Rho. PMID- 10362595 TI - Alternate splicing in human Na+-MI cotransporter gene yields differentially regulated transport isoforms. AB - myo-Inositol is a ubiquitous intracellular organic osmolyte and phosphoinositide precursor maintained at millimolar intracellular concentrations through the action of membrane-associated Na+-myo-inositol cotransporters (SMIT). Functional cloning and expression of a canine SMIT cDNA, which conferred SMIT activity in Xenopus oocytes, predicted a 718-amino acid peptide homologous to the Na+-glucose cotransporter with a potential protein kinase A phosphorylation site and multiple protein kinase C phosphorylation sites. A consistent approximately 1.0- to 13.5 kb array of transcripts hybridizing with this cDNA are osmotically induced in a variety of mammalian cells and species, yet SMIT activity appears to vary among different tissues and species. An open reading frame on human chromosome 21 (SLC5A3) homologous to that of the canine cDNA (96.5%) is thought to comprise an intronless human SMIT gene. Recently, this laboratory ascribed multiply sized, osmotically induced SMIT transcripts in human retinal pigment epithelial cells to the alternate utilization of several 3'-untranslated SMIT exons. This article describes an alternate splice donor site within the coding region that extends the open reading frame into the otherwise untranslated 3' exons, potentially generating novel SMIT isoforms. In these isoforms, the last putative transmembrane domain is replaced with intracellular carboxy termini containing a novel potential protein kinase A phosphorylation site and multiple protein kinase C phosphorylation sites, and this could explain the heterogeneity in the regulation and structure of the SMIT. PMID- 10362596 TI - Prepulse-induced mode 2 gating behavior with and without beta-adrenergic stimulation in cardiac L-type Ca channels. AB - Mode 2 gating of L-type Ca channels is characterized by high channel open probability (NPo) and long openings. In cardiac myocytes, this mode is evoked physiologically in two apparently different circumstances: membrane depolarization (prepulse facilitation) and activation of protein kinase A. To examine whether the phosphorylation mechanism is involved during prepulse-induced facilitation of cardiac L-type Ca channels, we used isolated guinea pig ventricular myocytes to analyze depolarization-induced modal gating behavior under different basal levels of phosphorylation. In control, NPo measured at 0 mV was augmented as the duration of prepulse to +100 mV was prolonged from 50 to 400 ms. This was due to the induction of mode 2 gating behavior clustered at the beginning of test pulses. Analysis of open time distribution revealed that the prepulse evoked an extra component, the time constant of which is not dependent on prepulse duration. When isoproterenol (1 microM) was applied to keep Ca channels at an enhanced level of phosphorylation, basal NPo without prepulse was increased by a factor of 3.6 +/- 2.2 (n = 6). Under these conditions, prepulse further increased NPo by promoting long openings with the same kinetics of transition to mode 2 gating (tau congruent with 200 ms at +100 mV). Likewise, recovery from mode 2 gating, as estimated by the decay of averaged unitary current, was not affected after beta-stimulation (tau congruent with 25 ms at 0 mV). The kinetic behavior independent from the basal level of phosphorylation or activity of cAMP-dependent protein kinase suggests that prepulse facilitation of the cardiac Ca channel involves a mechanism directly related to voltage-dependent conformational change rather than voltage-dependent phosphorylation. PMID- 10362597 TI - Effect of subunit composition and Liddle's syndrome mutations on biosynthesis of ENaC. AB - The epithelial Na+ channel (ENaC) is comprised of three homologous subunits: alpha, beta, and gamma, all of which are required for formation of the fully functional channel. This channel is responsible for salt reabsorption in the kidney, the airway, and the large bowel. Mutations in ENaC can cause human disease by increasing channel function in Liddle's syndrome, a form of hereditary hypertension, or by decreasing channel function in pseudohypoaldosteronism type I, a salt-wasting disease of infancy. We previously showed that ENaC is expressed on the cell surface as a minimally glycosylated, Triton-insoluble protein. In the present study we found that ENaC existed initially as a Triton-soluble protein that contained high-mannose glycosylation, presumably in the endoplasmic reticulum. This form of the protein disappeared as the Triton-insoluble, minimally glycosylated form became the more prevalent species. In pulse-chase studies of individually expressed subunits, we found that the Triton-soluble form of beta-ENaC accumulated initially, whereas the Triton-soluble form of alpha-ENaC decreased throughout the time course. However, when all three subunits were coexpressed, the alpha- and beta-subunits showed a similar pattern. The complex became Triton insoluble at some point after the endoplasmic reticulum, as incubation at 15 degrees C blocked the conversion to the insoluble form. Deletion of the carboxy-terminal tail of beta-ENaC causes Liddle's syndrome. This mutation increased the amount of newly synthesized Triton-insoluble ENaC heteromultimers but did not affect the half-life of insoluble protein. Therefore, subunit composition and mutations in individual subunits can influence biosynthesis of the ENaC complex. PMID- 10362598 TI - KGF prevents hyperoxia-induced reduction of active ion transport in alveolar epithelial cells. AB - We evaluated the effects of acute hyperoxic exposure on alveolar epithelial cell (AEC) active ion transport and on expression of Na+ pump (Na+-K+-ATPase) and rat epithelial Na+ channel subunits. Rat AEC were cultivated in minimal defined serum free medium (MDSF) on polycarbonate filters. Beginning on day 5, confluent monolayers were exposed to either 95% air-5% CO2 (normoxia) or 95% O2-5% CO2 (hyperoxia) for 48 h. Transepithelial resistance (Rt) and short-circuit current (Isc) were determined before and after exposure. Na+ channel alpha-, beta-, and gamma-subunit and Na+-K+-ATPase alpha1- and beta1-subunit mRNA levels were quantified by Northern analysis. Na+ pump alpha1- and beta1-subunit protein abundance was quantified by Western blotting. After hyperoxic exposure, Isc across AEC monolayers decreased by approximately 60% at 48 h relative to monolayers maintained under normoxic conditions. Na+ channel beta-subunit mRNA expression was reduced by hyperoxia, whereas alpha- and gamma-subunit mRNA expression was unchanged. Na+ pump alpha1-subunit mRNA was unchanged, whereas beta1-subunit mRNA was decreased approximately 80% by hyperoxia in parallel with a reduction in beta1-subunit protein. Because keratinocyte growth factor (KGF) has recently been shown to upregulate AEC active ion transport and expression of Na+-K+-ATPase under normoxic conditions, we assessed the ability of KGF to prevent hyperoxia-induced changes in active ion transport by supplementing medium with KGF (10 ng/ml) from day 2. The presence of KGF prevented the effects of hyperoxia on ion transport (as measured by Isc) relative to normoxic controls. Levels of beta1 mRNA and protein were relatively preserved in monolayers maintained in MDSF and KGF compared with those cultivated in MDSF alone. These results indicate that AEC net active ion transport is decreased after 48 h of hyperoxia, likely as a result of a decrease in the number of functional Na+ pumps per cell. KGF largely prevents this decrease in active ion transport, at least in part, by preserving Na+ pump expression. PMID- 10362599 TI - Is the chemical gate of connexins voltage sensitive? Behavior of Cx32 wild-type and mutant channels. AB - Connexin channels are gated by transjunctional voltage (Vj) or CO2 via distinct mechanisms. The cytoplasmic loop (CL) and arginines of a COOH-terminal domain (CT1) of connexin32 (Cx32) were shown to determine CO2 sensitivity, and a gating mechanism involving CL-CT1 association-dissociation was proposed. This study reports that Cx32 mutants, tandem, 5R/E, and 5R/N, designed to weaken CL-CT1 interactions, display atypical Vj and CO2 sensitivities when tested heterotypically with Cx32 wild-type channels in Xenopus oocytes. In tandems, two Cx32 monomers are linked NH2-to-COOH terminus. In 5R/E and 5R/N mutants, glutamates or asparagines replace CT1 arginines. On the basis of the intriguing sensitivity of the mutant-32 channel to Vj polarity, the existence of a "slow gate" distinct from the conventional Vj gate is proposed. To a lesser extent the slow gate manifests itself also in homotypic Cx32 channels. Mutant-32 channels are more CO2 sensitive than homotypic Cx32 channels, and CO2-induced chemical gating is reversed with relative depolarization of the mutant oocyte, suggesting Vj sensitivity of chemical gating. A hypothetical pore-plugging model involving an acidic cytosolic protein (possibly calmodulin) is discussed. PMID- 10362600 TI - Regulation of Spi 2.1 and 2.2 gene expression after turpentine inflammation: discordant responses to IL-6. AB - The rat serine protease inhibitor (Spi) 2 gene family includes both positive (Spi 2.2) and negative (Spi 2.1) acute phase reactants, facilitating modeling of regulation of hepatic acute phase response (APR). To examine the role of signal transducer and activation of transcription (STAT) proteins in the divergent regulation of these model genes after induction of APR, we evaluated the proximal promoters of the genes, focusing on STAT binding sites contained in these promoter elements. Induction of APR by turpentine injection includes activation of a STAT3 complex that can bind to a gamma-activated sequence (GAS) in the Spi 2.2 gene promoter, although the Spi 2.2 GAS site can bind STAT1 or STAT5 as well. To create an in vitro model of APR, primary hepatocytes were treated with combinations of cytokines and hormones to mimic the hormonal milieu of the whole animal after APR induction. Incubation of primary rat hepatocytes with interleukin (IL)-6, a critical APR cytokine, leads to activation of STAT3 and a 28-fold induction of a chloramphenicol acetyltransferase reporter construct containing the -319 to +85 region of the Spi 2.2 promoter. This suggests the turpentine-induced increase of Spi 2.2 is mediated primarily by IL-6. In contrast, although turpentine treatment reduces Spi 2.1 mRNA in vivo and IL-6 does not increase Spi 2.1 mRNA in primary rat hepatocytes, treatment of hepatocytes with IL-6 results in a 5. 4-fold induction of Spi 2.1 promoter activity mediated through the paired GAS elements in this promoter. Differential regulation of Spi 2.1 and 2.2 genes is due in part to differences in the promoters of these genes at the GAS sites. IL-6 alone fails to reproduce the pattern of rat Spi 2 gene expression that results from turpentine-induced inflammation. PMID- 10362601 TI - Vasoconstrictors and nitrovasodilators reciprocally regulate the Na+-K+-2Cl- cotransporter in rat aorta. AB - Little is known about the function and regulation of the Na+-K+-2Cl- cotransporter NKCC1 in vascular smooth muscle. The activity of NKCC1 was measured as the bumetanide-sensitive efflux of 86Rb+ from intact smooth muscle of the rat aorta. Hypertonic shrinkage (440 mosmol/kgH2O) rapidly doubled cotransporter activity, consistent with its volume-regulatory function. NKCC1 was also acutely activated by the vasoconstrictors ANG II (52%), phenylephrine (50%), endothelin (53%), and 30 mM KCl (54%). Both nitric oxide and nitroprusside inhibited basal NKCC1 activity (39 and 34%, respectively), and nitroprusside completely reversed the stimulation by phenylephrine. The phosphorylation of NKCC1 was increased by hypertonic shrinkage, phenylephrine, and KCl and was reduced by nitroprusside. The inhibition of NKCC1 significantly reduced the contraction of rat aorta induced by phenylephrine (63% at 10 nM, 26% at 30 nM) but not by KCl. We conclude that the Na+-K+-2Cl- cotransporter in vascular smooth muscle is reciprocally regulated by vasoconstrictors and nitrovasodilators and contributes to smooth muscle contraction, indicating that alterations in NKCC1 could influence vascular smooth muscle tone in vivo. PMID- 10362602 TI - TNF-alpha and insulin, alone and synergistically, induce plasminogen activator inhibitor-1 expression in adipocytes. AB - Obesity is associated with hyperinsulinemia and elevated concentrations of tumor necrosis factor-alpha (TNF-alpha) in adipose tissue. TNF-alpha has been implicated as an inducer of the synthesis of plasminogen activator inhibitor-1 (PAI-1), the primary physiological inhibitor of fibrinolysis, mediated by plasminogen activators in cultured adipocytes. To identify mechanism(s) through which TNF-alpha induces PAI-1, 3T3-L1 preadipocytes were differentiated into adipocytes and exposed to TNF-alpha for 24 h. TNF-alpha selectively increased the synthesis of PAI-1 without increasing activity of plasminogen activators. Both superoxide (generated by xanthine oxidase plus hypoxanthine) and hydrogen peroxide were potent inducers of PAI-1, and hydroxyl radical scavengers completely abolished the TNF-alpha induction of PAI-1. Exposure of adipocytes to TNF-alpha or insulin alone over 5 days increased PAI-1 production. These agonists exert synergistic effects. Results obtained suggest that TNF-alpha stimulates PAI 1 production by adipocytes, an effect potentiated by insulin, and that adipocyte generation of reactive oxygen centered radicals mediates the induction of PAI-1 production by TNF-alpha. Because induction of PAI-1 by TNF-alpha is potentiated synergistically by insulin, both agonists appear likely to contribute to the impairment of fibrinolytic system capacity typical in obese, hyperinsulinemic patients. PMID- 10362603 TI - Metabolic acidosis regulates rat renal Na-Si cotransport activity. AB - Recently, we cloned a cDNA (NaSi-1) localized to rat renal proximal tubules and encoding the brush-border membrane (BBM) Na gradient-dependent inorganic sulfate (Si) transport protein (Na-Si cotransporter). The purpose of the present study was to determine the effect of metabolic acidosis (MA) on Na-Si cotransport activity and NaSi-1 protein and mRNA expression. In rats with MA for 24 h (but not 6 or 12 h), there was a significant increase in the fractional excretion of Si, which was associated with a 2.4-fold decrease in BBM Na-Si cotransport activity. The decrease in Na-Si cotransport correlated with a 2.8-fold decrease in BBM NaSi-1 protein abundance and a 2.2-fold decrease in cortical NaSi-1 mRNA abundance. The inhibitory effect of MA on BBM Na-Si cotransport was also sustained in rats with chronic (10 days) MA. In addition, in Xenopus laevis oocytes injected with mRNA from kidney cortex, there was a significant reduction in the induced Na-Si cotransport in rats with MA compared with control rats, suggesting that MA causes a decrease in the abundance of functional mRNA encoding the NaSi-1 cotransporter. These findings indicate that MA reduces Si reabsorption by causing decreases in BBM Na-Si cotransport activity and that decreases in the expression of NaSi-1 protein and mRNA abundance, at least in part, play an important role in the inhibition of Na-Si cotransport activity during MA. PMID- 10362604 TI - Malignant human gliomas express an amiloride-sensitive Na+ conductance. AB - Human astrocytoma cells were studied using whole cell patch-clamp recording. An inward, amiloride-sensitive Na+ current was identified in four continuous cell lines originally derived from human glioblastoma cells (CH235, CRT, SKMG-1, and U251-MG) and in three primary cultures of cells obtained from glioblastoma multiforme tumors (up to 4 passages). In addition, cells freshly isolated from a resected medulloblastoma tumor displayed this same characteristic inward current. In contrast, amiloride-sensitive currents were not observed in normal human astrocytes, low-grade astrocytomas, or juvenile pilocytic astrocytomas. The only amiloride-sensitive Na+ channels thus far molecularly identified in brain are the brain Na+ channels (BNaCs). RT-PCR analyses demonstrated the presence of mRNA for either BNaC1 or BNaC2 in these tumors and in normal astrocytes. These results indicate that the functional expression of amiloride-sensitive Na+ currents is a characteristic feature of malignant brain tumor cells and that this pathway may be a potentially useful target for therapeutic intervention. PMID- 10362605 TI - Temporal responses of oxidative vs. glycolytic skeletal muscles to K+ deprivation: Na+ pumps and cell cations. AB - When K+ output exceeds input, skeletal muscle releases intracellular fluid K+ to buffer the fall in extracellular fluid (ECF) K+. To investigate the mechanisms and muscle specificity of the K+ shift, rats were fed K+-deficient chow for 2-10 days, and two muscles at phenotypic extremes were studied: slow-twitch oxidative soleus and fast-twitch glycolytic white gastrocnemius (WG). After 2 days of low K+ chow, plasma K+ concentration ([K+]) fell from 4.6 to 3.7 mM, and Na+-K+ ATPase alpha2 (not alpha1) protein levels in both muscles, measured by immunoblotting, decreased 36%. Cell [K+] decreased from 116 to 106 mM in soleus and insignificantly in WG, indicating that alpha2 can decrease before cell [K+]. After 5 days, there were further decreases in alpha2 (70%) and beta2 (22%) in WG, not in soleus, whereas cell [K+] decreased and cell [Na+] increased by 10 mM in both muscles. By 10 days, plasma [K+] fell to 2.9 mM, with further decreases in WG alpha2 (94%) and beta2 (70%); cell [K+] fell 19 mM in soleus and 24 mM in WG compared with the control, and cell [Na+] increased 9 mM in soleus and 15 mM in WG; total homogenate Na+-K+-ATPase activity decreased 19% in WG and insignificantly in soleus. Levels of alpha2, beta1, and beta2 mRNA were unchanged over 10 days. The ratios of alpha2 to alpha1 protein levels in both control muscles were found to be nearly 1 by using the relative changes in alpha-isoforms vs. beta1- (soleus) or beta2-isoforms (WG). We conclude that the patterns of regulation of Na+ pump isoforms in oxidative and glycolytic muscles during K+ deprivation mediated by posttranscriptional regulation of alpha2beta1 and alpha2beta2 are distinct and that decreases in alpha2-isoform pools can occur early enough in both muscles to account for the shift of K+ to the ECF. PMID- 10362606 TI - Differential activation of the SMalphaA promoter in smooth vs. skeletal muscle cells by bHLH factors. AB - E-box/basic helix-loop-helix (bHLH)-dependent regulation of promoters for skeletal muscle-specific genes is well established, but similar regulation of smooth muscle-selective promoters has not been reported. Using transient transfection assays of smooth muscle alpha-actin (SMalphaA) promoter chloramphenicol acetyltransferase (CAT) reporter constructs in rat vascular smooth muscle cells (SMCs) and L6 skeletal myotubes, we identified two activator elements, smE1 and smE2, with sequences corresponding to E-box (5'-CAnnTG-3') motifs. In L6 myotubes, 4-bp mutations of smE1 or smE2 E-box motif alone completely abolished promoter activity. In contrast, mutation of smE1 and smE2 was required to reduce promoter activity in SMCs. Supershift analyses identified a myogenin-containing complex as the predominant smE1 and smE2 binding activity in skeletal muscle, and myogenin overexpression transactivated the promoter. Supershift analyses with SMC extracts demonstrated that the bHLH protein upstream stimulatory factor (USF) bound smE1, and USF overexpression transactivated the promoter in an smE1-dependent manner. In summary, our results provide novel evidence implicating E-box elements in directing expression of the SMalphaA promoter through distinct bHLH factor complexes in skeletal vs. smooth muscle. PMID- 10362607 TI - Effect of coupling on volume-regulatory response of ciliary epithelial cells suggests mechanism for secretion. AB - The ciliary epithelium of the eye secretes the aqueous humor. It is a double epithelium arranged so that the apical surfaces of the nonpigmented ciliary epithelial (NPCE) and pigmented ciliary epithelial (PCE) cells face each other and the basolateral membranes face the inside of the eye and the blood, respectively. We have investigated the volume responses of both single cells and coupled pairs from this tissue to osmotic challenge. Both NPCE and PCE cells undergo regulatory volume increase (RVI) and decrease (RVD) when exposed to hyper and hyposmotic solution, respectively. In hyposmotic solution single cells swell and return to their original volumes within approximately 3 min. In nonpigmented cells RVD could be inhibited by blockers of volume-activated Cl- channels [tamoxifen (100%) > quinidine (87%) > DIDS (84%) > 5-nitro-2-(3 phenylpropylamino)benzoic acid (80%) > SITS (58%)] and K+ channels [Ba2+ (31%)]. However, in PCE cells these inhibitors and additionally tetraethylammonium and Gd3+ were without effect. Only bumetanide, an inhibitor of Na+-K+-2Cl- cotransport, was found to have any effect on RVD in PCE cells. NPCE-PCE cell coupled pairs also underwent RVD, but with altered kinetics. The onset of RVD of the PCE cell in a pair occurred approximately 80 s before that of the NPCE cell, and the peak swell was reduced. This is consistent with fluid movement from the PCE to the NPCE cell. The effect of the volume-activated Cl- channel inhibitor tamoxifen was to eliminate this difference in the times of onset of RVD in coupled cell pairs and to inhibit RVD in both the NPCE and PCE cells partially. On the basis of these observations we suggest that fluid is transferred from the PCE to the NPCE cell in coupled pairs during cell swelling and the subsequent RVD. Furthermore, we speculate that reciprocal RVI-RVD could underlie aqueous humor secretion. PMID- 10362608 TI - Inhibition of P-glycoprotein-mediated transport by a hydrophobic contaminant in commercial gluconate salts. AB - The substitution of gluconate for Cl- is commonly used to characterize Cl- transport or Cl--dependent transport mechanisms. We evaluated the effects of substituting gluconate for Cl- on the transport of the P-glycoprotein substrate rhodamine 123 (R123). The replacement of Ringer solution containing Cl- (Cl- Ringer) with gluconate-Ringer inhibited R123 efflux, whereas the replacement of Cl- by other anions (sulfate or cyclamate) had no effect. The inhibition of R123 efflux by gluconate-Ringer was absent after chloroform extraction of the sodium gluconate salt. The readdition of the sodium gluconate-chloroform extract to the extracted gluconate-Ringer or to cyclamate-Ringer inhibited R123 efflux, whereas its addition to Cl--Ringer had no effect. These observations indicate that the inhibition of P-glycoprotein-mediated R123 transport by gluconate is due to one or more chloroform-soluble contaminants and that the inhibition is absent in the presence of Cl-. The results are consistent with the fact that P-glycoprotein substrates are hydrophobic. Care should be taken when replacing ions to evaluate membrane transport mechanisms because highly pure commercial preparations may still contain potent contaminants that affect transport. PMID- 10362610 TI - Hydration of fat-free body mass: new physiological modeling approach. AB - Water is an essential component of living organisms, and in adult mammals the fraction of fat-free body mass (FFM) as water is remarkably stable at approximately 0.73. The stability of FFM hydration is a cornerstone of the widely used water isotope dilution method of estimating total body fat. At present, the only suggested means of studying FFM hydration is by experimental total body water (TBW) and FFM measurements. Although deviations from the classical hydration constant are recognized, it is unknown if these are explainable physiological aberrations and/or methodological errors. Moreover, many questions related to hydration stability prevail, including body mass and age effects. These unresolved questions and the importance of the TBW-fat estimation method led us to develop a cellular level FFM hydration model. This physiological model reveals that four water-related ratios combine to produce the observed TBW-to-FFM ratio. The mean and range of FFM hydration observed in adult humans can be understood with the proposed physiological model as can variation in the TBW-to FFM ratio over the human life span. An extension of the model to the tissue-organ body composition level confirms on a theoretical basis a small but systematic decrease in hydration observed in mammals ranging in body mass by a factor of 10(5). The present study, the first to advance a physiological hydration model, provides a conceptual framework for the TBW-fat estimation method and identifies important areas that remain to be studied. PMID- 10362609 TI - Molecular mechanism underlying a Cx50-linked congenital cataract. AB - Mutations in gap junctional channels have been linked to certain forms of inherited congenital cataract (D. Mackay, A. Ionides, V. Berry, A. Moore, S. Bhattacharya, and A. Shiels. Am. J. Hum. Genet. 60: 1474-1478, 1997; A. Shiels, D. Mackay, A. Ionides, V. Berry, A. Moore, and S. Bhattacharya. Am. J. Hum. Genet. 62: 526-532, 1998). We used the Xenopus oocyte pair system to investigate the functional properties of a missense mutation in the human connexin 50 gene (P88S) associated with zonular pulverulent cataract. The associated phenotype for the mutation is transmitted in an autosomal dominant fashion. Xenopus oocytes injected with wild-type connexin 50 cRNA developed gap junctional conductances of approximately 5 microS 4-7 h after pairing. In contrast, the P88S mutant connexin failed to form functional gap junctional channels when paired homotypically. Moreover, the P88S mutant functioned in a dominant negative manner as an inhibitor of human connexin 50 gap junctional channels when coinjected with wild type connexin 50 cRNA. Cells injected with 1:5 and 1:11 ratios of P88S mutant to wild-type cRNA exhibited gap junctional coupling of approximately 8% and 39% of wild-type coupling, respectively. Based on these findings, we conclude that only one P88S mutant subunit is necessary per gap junctional channel to abolish channel function. PMID- 10362611 TI - Neuronal actions of oxytocin on the subfornical organ of male rats. AB - The aim of this study was to investigate effects of oxytocin (OT) on electrical neuronal activities in rat subfornical organ (SFO) and compare its action with the well-described excitatory effects of blood-borne angiotensin II (ANG II) on the same SFO neurons. With the use of extracellular recordings from spontaneously active neurons in slice preparations of the SFO of male rats, 11.7% of tested neurons (n = 206) were excited and 9.7% were inhibited by superfusion with 10(-6) M OT. Both excitatory and inhibitory effects of OT were dose dependent with similar threshold concentrations and were blocked by a specific OT-receptor antagonist but not by a vasopressin receptor antagonist. Blocking synaptic transmission with low calcium medium suppressed only inhibitory effects of OT. All but one of the OT-sensitive neurons were also excited by superfusion with ANG II at a concentration much lower than required for OT, suggesting that synaptically released OT rather than blood-borne OT alters the activity of SFO neurons in vivo. The results support the hypothesis that neurally released OT may modulate SFO-mediated functions by acting on OT-sensitive neurons. PMID- 10362612 TI - Dose-response study of GH effects on circulating IGF-I and IGFBP-3 levels in healthy young men and women. AB - The aim of our study was to define the dose-response effect of a short-term treatment with different recombinant human growth hormone (rhGH) doses (1.25, 2.5, 5.0, 10.0, and 20.0 micrograms . kg-1. day-1 for 4 days) on insulin-like growth factor I (IGF-I) and insulin-like growth factor-binding protein (IGFBP)-3 levels in 21 normal young adults of both sexes. The dose of 1.25 microgram/kg rhGH did not modify IGF-I levels. The dose of 2.5 micrograms/kg rhGH significantly increased IGF-I levels in men (P < 0.05) but not in women, whereas the higher doses increased IGF-I levels in both sexes (P < 0.002). IGFBP-3 levels were not modified by 1.25 or 2.5 micrograms/kg rhGH in either sex. On the other hand, 5.0 micrograms/kg increased IGFBP-3 levels in men (P < 0.05) but not in women, whereas the higher doses increased IGFBP-3 levels similarly in both sexes (P < 0.02). In conclusion, our results demonstrate that IGF-I and IGFBP-3 responses to rhGH are dose and sex dependent. However, IGFBP-3 is less sensitive than IGF-I to rhGH stimulation. PMID- 10362613 TI - Protein kinetics during and after long-duration spaceflight on MIR. AB - Human spaceflight is associated with a loss of body protein. Bed rest studies suggest that the reduction in the whole body protein synthesis (PS) rate should be approximately 15%. The objectives of this experiment were to test two hypotheses on astronauts and cosmonauts during long-duration (>3 mo) flights on MIR: that 1) the whole body PS rate will be reduced and 2) dietary intake and the PS rate should be increased postflight because protein accretion is occurring. The 15N glycine method was used for measuring whole body PS rate before, during, and after long-duration spaceflight on the Russian space station MIR. Dietary intake was measured together with the protein kinetics. Results show that subjects lost weight during flight (4.64 +/- 1.0 kg, P < 0.05). Energy intake was decreased inflight (2,854 +/- 268 vs. 2,145 +/- 190 kcal/day, n = 6, P < 0.05), as was the PS rate (226 +/- 24 vs. 97 +/- 11 g protein/day, n = 6, P < 0.01). The reduction in PS correlated with the reduction in energy intake (r2 = 0.86, P < 0.01, n = 6). Postflight energy intake and PS returned to, but were not increased over, the preflight levels. We conclude that the reduction in PS found was greater than predicted from ground-based bed rest experiments because of the shortfall in dietary intake. The expected postflight anabolic state with increases in dietary intake and PS did not occur during the first 2 wk after landing. PMID- 10362614 TI - Prior exercise increases net hepatic glucose uptake during a glucose load. AB - The aim of these studies was to determine whether prior exercise enhances net hepatic glucose uptake (NHGU) during a glucose load. Sampling catheters (carotid artery, portal, hepatic, and iliac veins), infusion catheters (portal vein and vena cava), and Doppler flow probes (portal vein, hepatic and iliac arteries) were implanted. Exercise (150 min; n = 6) or rest (n = 6) was followed by a 30 min control period and a 100-min experimental period (3.5 mg. kg-1. min-1 of glucose in portal vein and as needed in vena cava to clamp arterial blood glucose at approximately 130 mg/dl). Somatostatin was infused, and insulin and glucagon were replaced intraportally at fourfold basal and basal rates, respectively. During experimental period the arterial-portal venous (a-pv) glucose gradient (mg/dl) was -18 +/- 1 in sedentary and -19 +/- 1 in exercised dogs. Arterial insulin and glucagon were similar in the two groups. Net hepatic glucose balance (mg. kg-1. min-1) shifted from 1.9 +/- 0.2 in control period to -1.8 +/- 0.2 (negative rates represent net uptake) during experimental period in sedentary dogs (Delta3.7 +/- 0.5); with prior exercise it shifted from 4.1 +/- 0.3 (P < 0.01 vs. sedentary) in control period to -3.2 +/- 0.4 (P < 0.05 vs. sedentary) during experimental period (Delta7.3 +/- 0.7, P < 0.01 vs. sedentary). Net hindlimb glucose uptake (mg/min) was 4 +/- 1 in sedentary animals in control period and 13 +/- 2 during experimental period; in exercised animals it was 7 +/- 1 in control period (P < 0. 01 vs. sedentary) and 32 +/- 4 (P < 0.01 vs. sedentary) during experimental period. As the total glucose infusion rate (mg. kg 1. min-1) was 7 +/- 1 in sedentary and 11 +/- 1 in exercised dogs, approximately 30% of the added glucose infusion due to prior exercise could be accounted for by the greater NHGU. In conclusion, when determinants of hepatic glucose uptake (insulin, glucagon, a-pv glucose gradient, glycemia) are controlled, prior exercise increases NHGU during a glucose load due to an effect that is intrinsic to the liver. Increased glucose disposal in the postexercise state is therefore due to an improved ability of both liver and muscle to take up glucose. PMID- 10362615 TI - Cytosolic citrate and malonyl-CoA regulation in rat muscle in vivo. AB - In liver, insulin and glucose acutely increase the concentration of malonyl-CoA by dephosphorylating and activating acetyl-CoA carboxylase (ACC). In contrast, in incubated rat skeletal muscle, they appear to act by increasing the cytosolic concentration of citrate, an allosteric activator of ACC, as reflected by increases in the whole cell concentrations of citrate and malate [Saha, A. K., D. Vavvas, T. G. Kurowski, A. Apazidis, L. A. Witters, E. Shafrir, and N. B. Ruderman. Am. J. Physiol. 272 (Endocrinol. Metab. 35): E641-E648, 1997]. We report here that sustained increases in plasma insulin and glucose may also increase the concentration of malonyl-CoA in rat skeletal muscle in vivo by this mechanism. Thus 70 and 125% increases in malonyl-CoA induced in skeletal muscle by infusions of glucose for 1 and 4 days, respectively, and a twofold increase in its concentration during a 90-min euglycemic-hyperinsulinemic clamp were all associated with significant increases in the sum of whole cell concentrations of citrate and/or malate. Similar correlations were observed in muscle of the hyperinsulinemic fa/fa rat, in denervated muscle, and in muscle of rats infused with insulin for 5 h. In muscle of 48-h-starved rats 3 and 24 h after refeeding, increases in malonyl-CoA were not accompanied by consistent increases in the concentrations of malate or citrate. However, they were associated with a decrease in the whole cell concentration of long-chain fatty acyl-CoA (LCFA-CoA), an allosteric inhibitor of ACC. The results suggest that increases in the concentration of malonyl-CoA, caused in rat muscle in vivo by sustained increases in plasma insulin and glucose or denervation, may be due to increases in the cytosolic concentration of citrate. In contrast, during refeeding after starvation, the increase in malonyl-CoA in muscle is probably due to another mechanism. PMID- 10362616 TI - Isradipine and insulin sensitivity in hypertensive rats. AB - The present study was designed to investigate the effect of a reduction in blood pressure, by using the calcium channel antagonist isradipine, on insulin sensitivity and vascular responses to insulin in conscious spontaneously hypertensive male rats (SHR). The rats were instrumented with intravascular catheters and pulsed Doppler flow probes to measure blood pressure, heart rate, and blood flows. Insulin sensitivity was assessed by the euglycemic hyperinsulinemic clamp technique. Two groups of rats received isradipine at a dose of 0.05 or 0.15 mg. kg-1. h-1, whereas a third group received a continuous infusion of vehicle (15% DMSO). Both doses of isradipine were found to decrease mean blood pressure (-25 +/- 4 mmHg at the dose of 0.05 mg. kg-1. h-1 and -20 +/- 2 mmHg at the dose of 0.15 mg. kg-1. h-1) and to improve insulin sensitivity. Moreover, in the rats treated with the low dose of isradipine, we observed vasodilations in renal, superior mesenteric, and hindquarter vascular beds. In the untreated group, the euglycemic infusion of insulin (4 mU. kg-1. min-1) was found to cause vasoconstrictions in superior mesenteric and hindquarter vascular beds, but no changes in mean blood pressure, heart rate, or renal vascular conductance were found. In contrast, in the isradipine-treated groups, the same dose of insulin was found to produce vasodilations in the renal vascular bed and to abolish the vasoconstrictor responses previously observed. We concluded that short-term treatment with isradipine in SHR can lower blood pressure and improve insulin sensitivity, mainly through hemodynamic factors, as supported by experiments with hydralazine as a positive vasodilator control. PMID- 10362617 TI - Effect of GIP and GLP-1 antagonists on insulin release in the rat. AB - Glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide 1 (GLP-1) are potent insulinotropic peptides released from the small intestine. To examine their relative contribution to postprandial insulin release, a specific GIP antagonist (ANTGIP) and a GLP-1 antagonist, exendin-(9-39)-NH2, were infused into rats after an intragastric glucose meal. In control rats, plasma glucose and insulin levels rose gradually during the first 20 min and then decreased. Exendin (9-39)-NH2 administration inhibited postprandial insulin secretion by 32% at 20 min and concomitantly increased plasma glucose concentrations. In contrast, ANTGIP treatment not only induced a 54% decrease in insulin secretion but also a 15% reduction in plasma glucose levels 20 min after the glucose meal. In vivo studies in rats demonstrated that glucose uptake in the upper small intestine was significantly inhibited by the ANTGIP, an effect that might account for the decrease in plasma glucose levels observed in ANTGIP-treated rats. When the two antagonists were administered to rats concomitantly, no potentiating effect on either insulin release or plasma glucose concentration was detected. Glucose meal stimulated GLP-1 release was not affected by ANTGIP administration, whereas postprandial glucagon levels were diminished in rats receiving exendin-(9-39) NH2. The results of these studies suggest that GIP and GLP-1 may share a common mechanism in stimulating pancreatic insulin release. Furthermore, the GIP receptor appears to play a role in facilitating glucose uptake in the small intestine. PMID- 10362618 TI - Acute enhancement of insulin secretion by FFA in humans is lost with prolonged FFA elevation. AB - The in vivo effect of elevated free fatty acids (FFA) on beta-cell function in humans remains extremely controversial. We examined, in healthy young men, the acute (90 min) and chronic (48 h) effects of an approximately twofold elevation of plasma FFA vs. control on glucose-stimulated insulin secretion (GSIS). GSIS was studied in response to a graded intravenous glucose infusion (peak plasma glucose, approximately 10 mmol/l, n = 8) and a two-step hyperglycemic clamp (10 and 20 mmol/l, n = 8). In the acute studies, GSIS was significantly higher, insulin sensitivity index (SI) was lower, and disposition index (DI = insulin sensitivity x insulin secretion) was unchanged with elevated FFA vs. control [2 step clamp: DI = 8.9 +/- 1.4 x 10(-3) l2. kg-1. min-2 in control vs. 10.0 +/- 1.9 x 10(-3) l2. kg-1. min-2 with high FFA, P = nonsignificant (NS)]. In the chronic studies, there was no difference in absolute GSIS between control and high FFA studies, but there was a reduction in SI and a loss of the expected compensatory increase in insulin secretion as assessed by the DI (2-step clamp: DI = 10.0 +/- 1.2 x 10(-3) l2. kg-1. min-2 in control vs. 6.1 +/- 0.7 x 10(-3) l2. kg-1. min-2 with high FFA, P = 0.01). In summary, 1) acute and chronic FFA elevation induces insulin resistance; 2) with acute FFA elevation, this insulin resistance is precisely countered by an FFA-induced increase in insulin secretion, such that DI does not change; and 3) chronic FFA elevation disables this beta-cell compensation. PMID- 10362619 TI - Mechanism of endothelin-1-(1-31)-induced calcium signaling in human coronary artery smooth muscle cells. AB - We have found that human chymase produces a 31-amino acid endothelin [ET-1-(1 31)] from the 38-amino acid precursor (Big ET-1). We examined the mechanism of synthetic ET-1-(1-31)-induced intracellular Ca2+ signaling in cultured human coronary artery smooth muscle cells. ET-1-(1-31) increased the intracellular free Ca2+ concentration ([Ca2+]i) in a concentration-dependent manner (10(-14)-10(-10) M). This ET-1-(1-31)-induced [Ca2+]i increase was not affected by phosphoramidon, Bowman-Birk inhibitor, and thiorphan. The ET-1-(1-31)-induced [Ca2+]i increase was not influenced by removal of extracellular Ca2+ but was inhibited by thapsigargin. ET-1-(1-31) at 10(-12) M did not cause Ca2+ influx, whereas 10(-7) M ET-1-(1-31) evoked marked Ca2+ influx, which was inhibited by nifedipine. ET-1 (1-31) also increased inositol trisphosphate formation. These results suggest that the ET-1-(1-31)-induced [Ca2+]i increase at relatively low concentrations is attributable to the release of Ca2+ from inositol trisphosphate-sensitive intracellular stores, whereas Ca2+ influx into the cells evoked by high concentration of ET-1-(1-31) probably occurs through voltage-dependent Ca2+ channels. We concluded that the physiological activity of ET-1-(1-31) may be attributable to Ca2+ mobilization from intracellular stores rather than influx of Ca2+ from extracellular space. PMID- 10362620 TI - Mitochondrial proton leak in brown adipose tissue mitochondria of Ucp1-deficient mice is GDP insensitive. AB - Mice deficient in mitochondrial uncoupling protein (UCP) 1 are cold sensitive, despite abundant expression of the homologues, Ucp2 and Ucp3 (S. Enerback, A. Jacobsson, E. M. Simpson, C. Guerra, H. Yamashita, M.-E. Harper, and L. P. Kozak. Nature 387: 90-94, 1997). We have analyzed characteristics of mitochondrial proton leak from brown adipose tissue (BAT) of Ucp1-deficient mice and normal controls and conducted the first top-down metabolic control analysis of oxidative phosphorylation in BAT mitochondria. Because purine nucleotides inhibit UCP1 and because UCP2 and the long form of UCP3 have putative purine nucleotide-binding regions, we predicted that proton leak in BAT mitochondria from Ucp1-deficient mice would be sensitive to GDP. On the contrary, although control over mitochondrial oxygen consumption and proton leak reactions at state 4 are strongly affected by 1 mM GDP in mitochondria from normal mice, there is no effect in UCP1-deficient mitochondria. In the presence of GDP, the overall kinetics of proton leak were not significantly different between Ucp1-deficient mice and controls. In its absence, state 4 respiration in normal BAT mitochondria was double that in its presence. Leak-dependent oxygen consumption was higher over a range of membrane potentials in its absence than in its presence. Thus proton leak, potentially including that through UCP2 and UCP3, is GDP insensitive. However, our measurements were made in the presence of albumin and may not allow for the detection of any fatty acid-induced UCP-mediated leak; this possibility requires investigation. PMID- 10362621 TI - A marathon run increases the susceptibility of LDL to oxidation in vitro and modifies plasma antioxidants. AB - Physical activity increases the production of oxygen free radicals, which may consume antioxidants and oxidize low-density lipoprotein (LDL). To determine whether this occurs during strenuous aerobic exercise, we studied 11 well-trained runners who participated in the Helsinki City Marathon. Blood samples were collected before, immediately after, and 4 days after the race to determine its effect on circulating antioxidants and LDL oxidizability in vitro. LDL oxidizability was increased as determined from a reduction in the lag time for formation of conjugated dienes both immediately after (180 +/- 7 vs. 152 +/- 4 min, P < 0.001) and 4 days after (155 +/- 7 min, P < 0.001) the race. No significant changes in lipid-soluble antioxidants in LDL or in the peak LDL particle size were observed after the race. Total peroxyl radical trapping antioxidant capacity of plasma (TRAP) and uric acid concentrations were increased after the race, but, except for TRAP, these changes disappeared within 4 days. Plasma thiol concentrations were reduced after the race. No significant changes were observed in plasma ascorbic acid, alpha-tocopherol, beta-carotene, and retinol concentrations after the marathon race. We conclude that strenuous aerobic exercise increases the susceptibility of LDL to oxidation in vitro for up to 4 days. Although the increase in the concentration of plasma TRAP reflects an increase of plasma antioxidant capacity, it seems insufficient to prevent the increased susceptibility of LDL to oxidation in vitro, which was still observed 4 days after the race. PMID- 10362622 TI - The acute-phase protein response to human immunodeficiency virus infection in human subjects. AB - Although several studies have shown that asymptomatic human immunodeficiency virus infection elicits an increase in whole body protein turnover, it is not known whether this increased protein turnover includes changes in the kinetics of acute-phase proteins (APPs). To answer this question, we measured 1) the plasma concentrations of four positive (C-reactive protein, alpha1-antitrypsin, haptoglobin, and fibrinogen) and four negative APPs [albumin, high-density lipoprotein (HDL)-apolipoprotein (apo) A1, transthyretin, and retinol-binding protein] and 2) the fractional (FSR) and absolute (ASRs) synthesis rates of three positive and three negative APPs using a constant intravenous infusion of [2H5]phenylalanine in five subjects with symptom-free acquired immunodeficiency syndrome (AIDS) and five noninfected control subjects. Compared with the values of the controls, the plasma concentrations, FSRs, and ASRs of most positive APPs were higher in the AIDS group. The negative APPs had faster FSRs in the AIDS group, there was no difference between the ASRs of the two groups, and only HDL apoA1 had a lower plasma concentration. These results suggest that symptom-free AIDS elicits an APP response that is different from bacterial infections, as the higher concentrations and faster rates of synthesis of the positive APPs are not accompanied by lower concentrations and slower rates of synthesis of most of the negative APPs. PMID- 10362623 TI - Passage of leptin across the blood-testis barrier. AB - Leptin is a 17-kDa protein, secreted by fat, that controls adiposity and has been proposed to have numerous effects on reproduction in the mouse. To assess whether the effects of leptin on testicular function are direct, we determined whether leptin can cross the murine blood-testis barrier. Multiple time regression analysis showed that a small amount of blood-borne leptin is able to enter the testis but does so by a nonsaturable process. In addition, no significant expression of leptin receptors was found at the Leydig cells or Sertoli cells of the testis. This compares with the presence of a saturable transport system for leptin at the blood-brain barrier and abundant receptors for leptin at the leptomeninges, neurons, and choroid plexus of the central nervous system (CNS). These results support the hypothesis that the effects of leptin on reproductive function are not mediated at the level of the testis but indirectly, probably through the CNS. PMID- 10362624 TI - Peripheral injection of growth hormone stimulates protein intake in aged male and female Lou rats. AB - It is well established that growth hormone (GH) induces growth rate and food efficiency and stimulates protein accretion in young mammals. Senescence is characterized by metabolic and hormonal disorders, particularly a decrease in protein turnover, which could be correlated to a decrease in GH and insulin-like growth factor I (IGF-I) secretion. We have shown that body weight, protein intake, and IGF-I plasma levels are greatly decreased with aging in Lou/C rats, particularly in males. In order to specify the GH effect on protein intake during aging, males and females (6, 19, and 24 mo) placed on a self-selection regimen were injected daily with a physiological dose of human GH (0.023 mg/rat sc). No GH effect on caloric intake was observed. Nevertheless, GH treatment stimulated body weight in older rats. It also increased protein intake in females and older males (19-24 mo). This stimulating effect was positively correlated with the degree of weight loss in senescent rats, suggesting that the decrease in protein intake observed with aging could be a marker of senescence. PMID- 10362625 TI - Estrogen receptors (ERalpha/ERbeta) in normal and pathological growth of the human myometrium: pregnancy and leiomyoma. AB - The distributions of the mRNAs for estrogen receptors (ERalpha and ERbeta) and their binding properties in myometria of pregnant and nonpregnant women and in leiomyoma were studied. RT-PCR analysis indicated that the term pregnancy myometria had little ERalpha mRNA, whereas the amounts of ERbeta mRNAs in pregnant or nonpregnant myometria appeared to be similar. Both ERalpha and ERbeta mRNA were greater in certain leiomyoma than in normal nonpregnant myometria. The binding kinetics revealed that two specific binding sites (with high or low affinity) for 17beta-estradiol were present in the nonpregnant myometrium. Only the low-affinity binding sites were detectable in late-pregnancy myometria and in leiomyoma, and their capacities were increased two- to threefold (P < 0.001) in leiomyoma. The pregnancy- and leiomyoma-related changes in myometrial ER status, especially the low concentration of ERalpha mRNA and the lack of high-affinity ER in pregnant women, plus the increased ERalpha and ERbeta mRNAs and the increased low-affinity ER in some leiomyoma, suggest that the redistribution of ER subtypes is associated with the pathological and/or normal growth of the myometrium. PMID- 10362626 TI - Effects of troglitazone on substrate storage and utilization in insulin-resistant rats. AB - Elevated serum and tissue lipid stores are associated with skeletal muscle insulin resistance and diminished glucose-stimulated insulin secretion, the hallmarks of type 2 diabetes. We studied the effects of 6-wk treatment with the insulin sensitizer troglitazone on substrate storage and utilization in lean control and Zucker diabetic fatty (ZDF) rats. Troglitazone prevented development of diabetes and lowered serum triglycerides (TG) in ZDF rats. Soleus muscle glycogen and TG content were elevated twofold in untreated ZDF rats, and both were normalized by troglitazone to lean control levels (P < 0.05). Troglitazone also normalized insulin-stimulated glucose uptake as well as basal and insulin stimulated glycogen synthesis, implying increased skeletal muscle glycogen turnover. The proportion of active pyruvate dehydrogenase (PDH) in soleus muscle was reduced in ZDF relative to lean control rat muscle (16 +/- 2 vs. 21 +/- 2%) but was restored by troglitazone treatment (30 +/- 3%). Increased PDH activation was associated with a 70% increase in glucose oxidation. Muscle lipoprotein lipase activity was decreased by 35% in ZDF compared with lean control rats and was increased twofold by troglitazone. Palmitate oxidation and incorporation into TG were higher in ZDF relative to lean control rats but were unaffected by troglitazone treatment. Troglitazone decreased the incorporation of glucose into the acyl group of TG by 60% in ZDF rats. In summary, ZDF rats demonstrate increased skeletal muscle glycogen and TG stores, both of which were reduced by troglitazone treatment. Troglitazone appears to increase both glycogen and TG turnover in skeletal muscle. Normalization of PDH activity and decreased glucose incorporation into acyl TG may underlie the improvements in intracellular substrate utilization and energy stores, which lead to decreased serum TG and glucose. PMID- 10362627 TI - Glucose production during strenuous exercise in humans: role of epinephrine. AB - The increase in hepatic glucose production (HGP) that occurs during intense exercise is accompanied by a simultaneous increase in epinephrine, which suggests that epinephrine may be important in regulating HGP. To further investigate this, six trained men were studied twice. The first trial [control (Con)] consisted of 20 min of cycling at 40 +/- 1% peak oxygen uptake (VO2 peak) followed by 20 min at 80 +/- 2% VO2 peak. During the second trial [epinephrine (Epi)], subjects exercised for 40 min at 41 +/- 2% VO2 peak. Epinephrine was infused during the latter 20 min of exercise and resulted in plasma levels similar to those measured during intense exercise in Con. Glucose kinetics were measured using a primed, continuous infusion of [3-3H]glucose. HGP was similar at rest (Con, 11.0 +/- 0.5 and Epi, 11.1 +/- 0.5 micromol. kg-1. min-1). In Con, HGP increased (P < 0.05) during exercise to 41.0 +/- 5.2 micromol. kg-1. min-1 at 40 min. In Epi, HGP was similar to Con during the first 20 min of exercise. Epinephrine infusion increased (P < 0.05) HGP to 24.0 +/- 2.5 micromol. kg-1. min-1 at 40 min, although this was less (P < 0.05) than the value in Con. The results suggest that epinephrine can increase HGP during exercise in trained men; however, epinephrine during intense exercise cannot fully account for the rise in HGP. Other glucoregulatory factors must contribute to the increase in HGP during intense exercise. PMID- 10362628 TI - Glutamine synthetase expression in muscle is regulated by transcriptional and posttranscriptional mechanisms. AB - Skeletal muscle exports glutamine (Gln) and increases the expression of the enzyme glutamine synthetase (GS) in response to physiological stress. Acute stress or direct glucocorticoid administration raises muscle GS mRNA levels dramatically without a parallel increase in GS protein levels. In the lung, this discrepancy is caused by feedback destabilization of the GS protein by its product Gln. It was hypothesized that muscle GS protein levels increase during stress only when the intracellular Gln pool has been depleted. Adult male rats were injected with the glucocorticoid hormone dexamethasone (DEX) to mimic the acute stress response and with the GS inhibitor methionine sulfoximine (MSO) to deplete muscle Gln stores. DEX increased GS mRNA levels by 2.8-fold but increased GS protein levels by an average of only 40%. MSO diminished muscle GLN levels by 68% and caused GS protein levels to rise in accordance with GS mRNA. Chronic stress was mimicked using 6 days of MSO treatment, which produced anorexia, 23% loss of body weight, and 64% decrease in muscle Gln levels, as well as pronounced increases in both muscle GS mRNA (26-fold) and protein levels (35-fold) without elevation of plasma glucocorticoid levels. Calorie-restricted pair-fed animals exhibited lesser increases in muscle GS mRNA (8-fold) and protein levels (5-fold) without a decline in muscle Gln content. Thus regulation of GS expression in both acute and chronic stress involved both transcriptional and posttranscriptional mechanisms, perhaps affected by muscle Gln content. PMID- 10362629 TI - Mass isotopomer distribution analysis at eight years: theoretical, analytic, and experimental considerations. AB - Mass isotopomer distribution analysis (MIDA) is a technique for measuring the synthesis of biological polymers. First developed approximately eight years ago, MIDA has been used for measuring the synthesis of lipids, carbohydrates, and proteins. The technique involves quantifying by mass spectrometry the relative abundances of molecular species of a polymer differing only in mass (mass isotopomers), after introduction of a stable isotope-labeled precursor. The mass isotopomer pattern, or distribution, is analyzed according to a combinatorial probability model by comparing measured abundances to theoretical distributions predicted from the binomial or multinomial expansion. For combinatorial probabilities to be applicable, a labeled precursor must therefore combine with itself in the form of two or more repeating subunits. MIDA allows dilution in the monomeric (precursor) and polymeric (product) pools to be determined. Kinetic parameters can then be calculated (e.g., replacement rate of the polymer, fractional contribution from the endogenous biosynthetic pathway, absolute rate of biosynthesis). Several issues remain unresolved, however. We consider here the impact of various deviations from the simple combinatorial probability model of biosynthesis and describe the analytic requirements for successful use of MIDA. A formal mathematical algorithm is presented for generating tables and equations (APPENDIX), on the basis of which effects of various confounding factors are simulated. These include variations in natural isotope abundances, isotopic disequilibrium in the precursor pool, more than one biosynthetic precursor pool, incorrect values for number of subunits present, and concurrent measurement of turnover from exogenously labeled polymers. We describe a strategy for testing whether isotopic inhomogeneity (e.g., an isotopic gradient or separate biosynthetic sites) is present in the precursor pool by comparing higher-mass (multiply labeled) to lower-mass (single- and double-labeled) isotopomer patterns. Also, an algebraic correction is presented for calculating fractional synthesis when an incomplete ion spectrum is monitored, and an approach for assessing the sensitivity of biosynthetic parameters to measurement error is described. The different calculation algorithms published for MIDA are compared; all share a common model, use overlapping solutions to computational problems, and generate identical results. Finally, we discuss the major practical issue for using MIDA at present: quantitative inaccuracy of instruments. The nature and causes of analytic inaccuracy, strategies for evaluating instrument performance, and guidelines for optimizing accuracy and reducing impact on biosynthetic parameters are suggested. Adherence to certain analytic guidelines, particularly attention to concentration effects on mass isotopomer ratios and maximizing enrichments in the isotopomers of interest, reduces error. Improving instrument accuracy for quantification of isotopomer ratios is perhaps the highest priority for this field. In conclusion, MIDA remains the "equation for biosynthesis," but attention to potentially confounding factors and analytic performance is required for optimal application. PMID- 10362630 TI - Undermodeling affects minimal model indexes: insights from a two-compartment model. AB - The classic (hereafter cold) and the labeled (hereafter hot) minimal models are powerful tools to investigate glucose metabolism. The cold model provides, from intravenous glucose tolerance test (IVGTT) data, indexes of glucose effectiveness (SG) and insulin sensitivity (SI) that measure the effect of glucose and insulin, respectively, to enhance glucose disappearance and inhibit endogenous glucose production. The hot model provides, from hot IVGTT data, indexes of glucose effectiveness (SG*) and insulin sensitivity (SI*) that, respectively, measure the effects of glucose and insulin on glucose disappearance only. Recent reports call for a reexamination of some of the assumptions of the minimal models. We have previously pointed out the criticality of the single-compartment description of glucose kinetics on which both the minimal models are founded. In this paper we evaluate the impact of single-compartment undermodeling on SG, SI*, and by using a two-compartment model to describe the glucose system. The relationships of the minimal model indexes to the analogous indexes measured with the glucose clamp technique are also examined. Theoretical analysis and simulation studies indicate that cold indexes are more affected than hot indexes by undermodeling. In particular, care must be exercised in the physiological interpretation of SG, because this index is a local descriptor of events taking place in the initial portion of the IVGTT. As a consequence, SG not only reflects glucose effect on glucose uptake and production but also the rapid exchange of glucose between the accessible and nonaccessible glucose pools that occurs in the early part of the test. PMID- 10362631 TI - Whole body protein kinetics using Phe and Tyr tracers: an evaluation of the accuracy of approximated flux values. AB - Phenylalanine (Phe) kinetics are increasingly used in studies of amino acid kinetics, because the metabolic fate of Phe is limited to incorporation into protein (protein synthesis, Sp) and catabolism via hydroxylation (Qpt) to tyrosine (Tyr). Besides an infusion of labeled Phe to measure Phe flux (Qp), a priming dose of Tyr and an independent Tyr tracer are used to measure Tyr flux (Qt) and Qpt. Alternatively, Qt, Qpt, and Sp can be approximated by using equations, based on Phe and Tyr concentrations in body proteins, that eliminate the need for a Tyr tracer. To evaluate the accuracy of this approach, data were obtained from 12 type I diabetic patients and 24 nondiabetic control subjects who were studied with the full complement of tracers both with and without insulin infusion. Sp approximations closely matched measured values in both groups (mean difference <2%, all values <5%), but the agreement was poor for Qpt (error range = -8 to +43%) and Qt (error range -22 to +41%). Insulin status had no effect on these comparisons. The lower approximation error for Sp vs. Qpt is due to the small contribution ( approximately 10%) of Qpt to Qp. Approximation error for Qpt (r > 0.99) can be explained by variability in the ratio of Tyr to Phe coming from protein breakdown, (Qt - Qpt)/Qp. Ideally, all fluxes should be directly measured, but these data suggest that whole body Sp can be approximated with an acceptably small margin of error. However, the same equations do not yield reliably accurate values for Qpt or Qt. PMID- 10362633 TI - Nitric Oxide. V. therapeutic potential of nitric oxide donors and inhibitors. AB - Nitric oxide is a crucial mediator of gastrointestinal mucosal defense, but, paradoxically, it also contributes to mucosal injury in several situations. Inhibitors of nitric oxide synthesis and compounds that release nitric oxide have been useful pharmacological tools for evaluating the role of nitric oxide in gastrointestinal physiology and pathophysiology. Newer inhibitors with selectivity for one of the isoforms of nitric oxide synthase are even more powerful tools and may have utility as therapeutic agents. Also, agents that can scavenge nitric oxide or peroxynitrite are promising as drugs to prevent nitric oxide-associated tissue injury. Compounds that release nitric oxide in small amounts over a prolonged period of time may also be very useful for prevention of gastrointestinal injury associated with shock and with the use of drugs that have ulcerogenic effects. Indeed, the coupling of a nitric oxide-releasing moiety to nonsteroidal anti-inflammatory drugs has proven to be a valid means of substantially reducing the gastrointestinal toxicity of these drugs without decreasing their efficacy. PMID- 10362634 TI - Mucosal immunity and inflammation. II. The yin and yang of T cells in intestinal inflammation: pathogenic and protective roles in a mouse colitis model. AB - Inflammatory bowel disease (IBD) is a multifactorial immune disorder of uncertain etiology. The advent of several mouse models of mucosal inflammation that resemble IBD has provided insight into the mechanisms governing both normal and pathological mucosal immune function. In a widely used adoptive transfer model, the injection into immunodeficient mice of a subset of CD4(+) T lymphocytes, the CD4(+)CD45RBhigh cells, leads to inflammation of the intestine. Pathogenesis is due in part to the secretion of proinflammatory cytokines. The induction of colitis can be prevented by cotransfer of another CD4(+) subpopulation, the CD4(+)CD45RBlow T cells. This population behaves analogously to the CD4(+)CD45RBhigh population in terms of the acquisition of activation markers and homing to the host intestine. However, their lymphokine profile when activated is different, and anti-inflammatory cytokines secreted and/or induced by CD4(+)CD45RBlow T cells prevent colitis. In this themes article, a description of the adoptive transfer model is given, the factors that promote and prevent colitis pathogenesis are discussed, and some controversial aspects of the model are addressed. PMID- 10362635 TI - Effect of heat stress on rabbit esophageal epithelium. AB - Hot beverages expose the esophageal epithelium to temperatures as high as 58 degrees C. To study the impact of such temperatures, rabbit esophageal epithelium was exposed to luminal heat or both luminal and serosal heat while mounted in Ussing chambers. Luminal heat, mimicking exposure to hot beverages, reduced potential difference (PD) and resistance (R) when applied at >/=49 degrees C and reduced short-circuit current (Isc) at >/=60 degrees C. At >/=60 degrees C, subepithelial blisters developed. Higher temperatures reduced R only moderately and reversibly. In contrast, the Isc declined sharply and irreversibly once threshold was reached. Luminal and serosal heat also reduced PD, Isc, and R, although the threshold for reduction in Isc was now similar to that for R. Additionally, luminal and serosal heat reduced Isc more than R for any given temperature and resulted in blisters at lower temperatures (50 degrees C) than luminal heat alone. The heat-induced decline in Isc was attributed in part to inactivation of Na-K-ATPase activity, although other transport systems could have been equally affected, and the decline in R to an increase in paracellular permeability. The latter effect on R also contributed to an increase in tissue sensitivity to luminal acid damage. Consumption of hot beverages exposes the esophagus to temperatures that can negatively impact epithelial structure and function. Impaired barrier function by heat increases the risk of esophageal damage by subsequent contact with (refluxed) gastric acid. These findings help explain in part the association between esophageal disease and consumption of hot beverages. PMID- 10362636 TI - Ca-sensitive Na transport in sheep omasum. AB - Na transport across a preparation of sheep omasum was studied. All tissues exhibited a serosa-positive short-circuit current (Isc), with a range of 1-4 microeq. h-1. cm-2. A Michaelis-Menten-type kinetic was found between the Na concentration and the Isc (Michaelis-Menten constant for transport of Na = 6.7 mM; maximal transport capacity of Na = 4.16 microeq. h-1. cm-2). Mucosal amiloride (1 mM), phenamil (1 or 10 microM), or serosal aldosterone (1 microM for 6 h) did not change Isc. Removal of divalent cations (Ca and Mg) enhanced Isc considerably from 2.61 +/- 0.24 to a peak value of 11.18 +/- 1.1 microeq. h-1. cm 2. The peak Isc (overshoot) immediately declined to a plateau Isc of approximately 6-7 microeq. h-1. cm-2. Na flux measurements showed a close correlation between changes in Isc and Na transport. Transepithelial studies demonstrated that K, Cs, Rb, and Li are transported, indicating putative nonselective cation channels, which are inhibited by divalent cations (including Ca, Mg, Sr, Ba) and by (trivalent) La. Intracellular microelectrode recordings from the luminal side clearly showed changes of voltage divider ratio when mucosal divalent cations were removed. The obtained data support the assumption of a distinct electrogenic Na transport mechanism in sheep omasum. PMID- 10362637 TI - Activation of VEGF and Ras genes in gastric mucosa during angiogenic response to ethanol injury. AB - Our previous studies demonstrated that ethanol injury triggers the angiogenic response in gastric mucosa bordering necrosis. The present study was aimed to determine whether vascular endothelial growth factor (VEGF) (a potent angiogenic peptide selectively acting on endothelial cells) and Ras (a mediator of cell proliferation and a putative regulator of VEGF expression) are involved in gastric angiogenesis after ethanol injury. We studied the angiogenic response and expression of VEGF and Ras in gastric mucosa after ethanol injury. Ethanol damage triggered 1) angiogenesis in the gastric mucosa bordering necrosis, 2) significant increases in VEGF mRNA and protein expression, and 3) significant increases in the expression of Ki-ras mRNA and Ras proteins. Neutralizing anti VEGF antibody significantly reduced (by greater than threefold) the angiogenic response to ethanol-induced injury. Moreover, mevastatin, an inhibitor of Ras activation, completely blocked the induction of VEGF expression in cultured primary endothelial cells. Because, in other tissues, VEGF is one of the most potent angiogenic factors and VEGF expression is dependent on Ras, our data indicate that Ras and VEGF are involved in gastric mucosal angiogenesis after ethanol injury. PMID- 10362638 TI - Roles of IL-1 and TNF in the decreased ileal muscle contractility induced by lipopolysaccharide. AB - Gastrointestinal stasis during sepsis may be associated with gastrointestinal smooth muscle dysfunction. Endotoxin [lipopolysaccharide (LPS)] impairs smooth muscle contraction, in part through inducible nitric oxide synthase (NOS II) and enhanced nitric oxide production. We studied the roles of tumor necrosis factor alpha (TNF) and interleukin-1 (IL-1) in this process by using TNF binding protein (TNFbp) and IL-1 receptor antagonist (IL-1ra). Rats were treated with TNFbp and IL-1ra, or their vehicles, 1 h before receiving LPS or saline. At 5 h after LPS, contractility was measured in strips of ileal longitudinal smooth muscle, and NOS II activity was measured in full-thickness segments of ileum. LPS decreased maximum stress (mean +/- SE) from 508 +/- 55 (control) to 355 +/- 33 g/cm2 (P < 0.05). Pretreatment with TNFbp plus IL-1ra prevented the LPS-induced decrease. Separate studies of TNFbp alone or IL-1ra alone indicated that, at the doses and timing used, TNFbp was more effective. LPS also increased NOS II activity by >10 fold (P < 0.01) over control. This increase was prevented by TNFbp plus IL-1ra (P = not significant vs. control). We conclude that the LPS-induced increase in NOS II activity and the decrease in ileal muscle contractility are mediated by TNF and IL-1. PMID- 10362639 TI - Cell type-specific requirement of the MAPK pathway for the growth factor action of gastrin. AB - Gastrin (G17) has a CCKB receptor-mediated growth-promoting effect on the AR42J rat acinar cell line that is linked to induction of both mitogen-activated protein kinase (MAPK) and c-fos gene expression. We investigated the mechanisms that regulate the growth factor action of G17 on the rat pituitary adenoma cell line GH3. Both AR42J and GH3 cells displayed equal levels of CCKB receptor expression and similar binding kinetics of 125I-labeled G17. G17 stimulation of cell proliferation was identical in both cell lines. G17 stimulation of GH3 cell proliferation was completely blocked by the CCKB receptor antagonist D2 but not by the MEK inhibitor PD-98059 or the protein kinase C inhibitor GF-109203X, which completely inhibited G17 induction of AR42J cell proliferation. G17 induced a c fos SRE-luciferase reporter gene plasmid more than fourfold in the AR42J cells, whereas it had no effect in the GH3 cells. In contrast to what we observed in the AR42J cells, G17 failed to stimulate MAPK activation and Shc tyrosyl phosphorylation and association with the adapter protein Grb2. Epidermal growth factor induced the MAPK pathway in the GH3 cells, demonstrating the integrity of this signaling system. G17 induced Ca2+ mobilization in both the GH3 and AR42J cells. The calmodulin inhibitor N-(6-aminohexyl)-5-chloro-1 naphthalenesulfonamide inhibited AR42J cell proliferation by 20%, whereas it completely blocked G17 induction of GH3 cell growth. The Ca2+ ionophore ionomycin stimulated GH3 cell proliferation to a level similar to that observed in response to G17, but it had no effect on AR42J cell proliferation. Thus there are cell type specific differences in the requirement of the MAPK pathway for the growth factor action of G17. Whereas in the AR42J cells G17 stimulates cell growth through activation of MAPK and c-fos gene expression, in the GH3 cells, G17 fails to activate MAPK, and it induces cell proliferation through Ca2+-dependent signaling pathways. Furthermore, induction of Ca2+ mobilization in the AR42J cells appears not to be sufficient to sustain cell proliferation. PMID- 10362640 TI - ATP is a mediator of the fast inhibitory junction potential in human jejunal circular smooth muscle. AB - The neurotransmitter(s) that generates the fast component of the inhibitory junction potential (IJP-F) in human jejunal circular smooth muscle is not known. The aim of this study was to determine the role of ATP and purinergic receptors in the generation of the IJP-F in human jejunal circular smooth muscle strips. The P2-receptor antagonist suramin (100 microM) reduced the IJP-F by 28%. Apamin (1 microM) reduced the IJP-F by 25%. Desensitization of muscle strips with the putative P2x-receptor agonist alpha, beta-methylene ATP (alpha,beta-MeATP, 100 microM) decreased the IJP-F by 44%, and desensitization with the putative P2y receptor agonist adenosine 5'-O-2-thiodiphosphate (ADPbetaS) completely abolished the IJP-F. Desensitization with the putative P2y-receptor agonist 2-methylthioATP had no effect on the IJP-F. Exogenous ATP evoked a hyperpolarization with a time course that matched the IJP-F. The ATP-evoked hyperpolarization was reduced by apamin and suramin, reduced by desensitization with alpha,beta-MeATP (69% decrease), and abolished by desensitization with ADPbetaS. These data suggest that the IJP-F in human jejunal circular smooth muscle is mediated in part by ATP through an ADPbetaS-sensitive P2 receptor. PMID- 10362641 TI - Uptake of bromosulfophthalein via SO2-4/OH- exchange increases the K+ conductance of rat hepatocytes. AB - In confluent primary cultures of rat hepatocytes, micromolar concentrations of bromosulfophthalein (BSP) lead to a sizeable hyperpolarization of membrane voltage. The effect is a saturable function of BSP concentration yielding an apparent value of 226 micromol/l and a Vmax of -10.3 mV. The BSP-induced membrane hyperpolarization is inhibited by the K+ channel blocker Ba2+, and in cable analysis and ion-substitution experiments it becomes evident that the effect is due to a significant increase in cell membrane K+ conductance. Voltage changes were attenuated by the simultaneous administration of SO2-4, succinate, and cholate (cis-inhibition) and increased after preincubation with SO2-4 and succinate (trans-stimulation), suggesting that the effect occurs via BSP uptake through the known SO2-4/OH- exchanger. Microfluorometric measurements reveal that BSP-induced activation of K+ conductance is not mediated by changes in cell pH, cell Ca2+, or cell volume. It is concluded that K+ channel activation by BSP (as well as by DIDS and indocyanine green) may reflect a physiological mechanism linking the sinusoidal uptake of certain anions to their electrogenic canalicular secretion. PMID- 10362642 TI - Endogenous ATP release regulates Cl- secretion in cultured human and rat biliary epithelial cells. AB - P2Y receptor stimulation increases membrane Cl- permeability in biliary epithelial cells, but the source of extracellular nucleotides and physiological relevance of purinergic signaling to biliary secretion are unknown. Our objectives were to determine whether biliary cells release ATP under physiological conditions and whether extracellular ATP contributes to cell volume regulation and transepithelial secretion. With the use of a sensitive bioluminescence assay, constitutive ATP release was detected from human Mz-ChA-1 cholangiocarcinoma cells and polarized normal rat cholangiocyte monolayers. ATP release increased rapidly during cell swelling induced by hypotonic exposure. In Mz-ChA-1 cells, removal of extracellular ATP (apyrase) and P2 receptor blockade (suramin) reversibly inhibited whole cell Cl- current activation and prevented cell volume recovery during hypotonic stress. Moreover, exposure to apyrase induced cell swelling under isotonic conditions. In intact normal rat cholangiocyte monolayers, hypotonic perfusion activated apical Cl- currents, which were inhibited by addition of apyrase and suramin to bathing media. These findings indicate that modulation of ATP release by the cellular hydration state represents a potential signal coordinating cell volume with membrane Cl- permeability and transepithelial Cl- secretion. PMID- 10362644 TI - Effects of sustained low-flow perfusion on the response to vasoconstrictor agents in postnatal intestine. AB - This laboratory has previously reported that sustained reduction of blood flow in newborn intestine causes a triphasic increase in vascular resistance that occurs over 3-4 h and that these changes are mediated, in part, by loss of endothelial nitric oxide (NO) production. This study examines the effects of exposure to sustained low-flow perfusion on the subsequent response to three contractile agonists: ANG II, norepinephrine (NE), and endothelin-1 (ET-1). Gut loops from 3- and 35-day-old swine were exposed to low-flow conditions in vivo (i.e., reduction of flow to approximately 50% of baseline) for 30 min or 5 h. Thereafter, they were removed to an extracorporeal perfusion circuit for in vitro hemodynamic assessment; alternatively, the mesenteric artery perfusing the gut loop was removed and cut into rings for assessment of isometric tension development. Gut loops from 3-day-old subjects exposed to low-flow conditions demonstrated significantly increased contractile responses to ANG II, NE, and ET-1; also, mesenteric artery rings from these gut loops demonstrated a significant reduction of the ED50 for all three agonists. Similar changes were not observed in intestine or mesenteric artery rings from older subjects. Sustained blockade of endogenous NO synthesis with NG-monomethyl- L-arginine duplicated the effects of exposure to sustained low-flow perfusion. It appears that sustained reduction of blood flow in newborn intestine decreases constitutive NO production, which in turn causes a generalized enhancement of the contractile efficacy of ANG II, NE, and ET-1. PMID- 10362643 TI - CCK receptor dysfunction in muscle membranes from human gallbladders with cholesterol stones. AB - Human gallbladders with cholesterol stones exhibit impaired muscle contraction induced by agonists that act on transmembrane receptors, increased membrane cholesterol content, and abnormal cholesterol-to-phospholipid ratio compared with those with pigment stones. The present study was designed to investigate the functions of the CCK receptor of gallbladder muscle membranes by radioreceptor assay and cross-linking. 125I-labeled CCK-8 binding was time-dependent, competitive, and specific. Scatchard analysis showed that the maximum specific binding (Bmax) was significantly decreased in cholesterol compared with pigment stone gallbladders (0.18 +/- 0. 07 vs. 0.38 +/- 0.05 pmol/mg protein, P < 0.05). In contrast, the affinity for CCK was higher in cholesterol than pigment stone gallbladders (0.18 +/- 0.06 vs. 1.2 +/- 0.23 nM). Similar results were observed in binding studies with the CCK-A receptor antagonist [3H]L-364,718. Cross linking and saturation binding studies also showed significantly less CCK binding in gallbladders with cholesterol stones. These abnormalities were reversible after incubation with cholesterol-free liposomes. The Bmax increased (P < 0.01) and the dissociation constant decreased (P < 0.001) after incubation with cholesterol-free liposomes. In conclusion, human gallbladders with cholesterol stones have impaired CCK receptor binding compared with those with pigment stones. These changes are reversed by removal of the excess membrane cholesterol. These receptor alterations may contribute to the defective contractility of the gallbladder muscle in patients with cholesterol stones. PMID- 10362645 TI - Visual parameters define the phase and the load of contractions in isolated guinea pig ileum. AB - How the movements of the intestinal walls relate to luminal pressures and outflow remains incompletely understood. We triggered the peristaltic reflex in the isolated ileum of the guinea pig and quantified wall movements through computerized measurements of diameter changes. Contractions developed as indentations close to the upstream end of the loop. The indentations deepened and expanded in length. The downstream shoulder of contractions started and stopped to propagate before the upstream shoulder. Shoulders differed in their length and gradient over most of the duration of the contraction, and this gives the contraction an axial asymmetry. Over the course of individual contractions, the length of the indented segment correlated well with the luminal pressure. Contractions in response to large volumes generated long indented segments and high luminal pressures. The onset and the end of pressure waves and of outflow did not necessarily coincide with the onset and end of visual parameters of contractions. These findings indicate that objective visual parameters might be useful to describe and to classify contractions. PMID- 10362646 TI - Transcriptional control of the murine polymeric IgA receptor promoter by glucocorticoids. AB - Glucocorticoids have been implicated as an important regulator of intestinal epithelial cell ontogeny. The polymeric IgA receptor (pIgR) is expressed in the intestinal epithelial layer and is regulated by several mediators, including glucocorticoids. The mechanism of how corticosteroids alter the transcriptional regulation of pIgR expression has not been defined. In this study, we demonstrated that glucocorticoids upregulate steady-state pIgR mRNA levels in the proximal intestine of suckling rats and in the IEC-6 intestinal cell line. We performed functional analysis of the 5'-flanking region in the presence of glucocorticoids and its receptor using the intestinal cell line Caco-2. We screened 4.7 kb of the upstream region of the murine gene and identified the most potent steroid response element to reside between nt -215 and -163 relative to the start of transcription. Substitution mutation analysis of this region identified the location of the putative steroid response element to be between nt -195 and -176. In vitro DNase I footprint analysis using the recombinant glucocorticoid receptor DNA binding domain confirmed a single area of protection that spans the nt identified by mutagenesis analysis. Electrophoretic mobility shift assays of the putative element confirmed the binding of both recombinant and cell synthesized glucocorticoid receptor in a specific manner. In summary, we report the identification and characterization of the glucocorticoid-DNA response element located in the immediate 5'-upstream region of the murine pIgR gene. PMID- 10362648 TI - Modeling intermittent digesta flow to calculate glucose uptake capacity of the bovine small intestine. AB - To test the hypothesis that the uptake capacity of the bovine small intestine for glucose is upregulated to match or slightly exceed glucose delivery, glucose was continuously infused into the proximal duodenum of four cannulated holstein heifers. Every 3 days, infusion rates were increased by an average of 34 mmol/h. A model of glucose disappearance from multiple boluses of intestinal digesta was used to estimate the transporter maximum velocity and functional maximum uptake capacity for the entire small intestine from average ileal glucose flows during the third day of each period. Because of its intermittency, digesta flow remained independent of simulated transit time. For each unit increase in glucose infusion rate, uptake capacity increased by only 0.55 units. Excess capacity for glucose uptake was approximately twofold in forage-fed cattle and declined to below delivery at infusions of >208 mmol/h added glucose, approximately three times the normal load. Calculations for cattle, sheep, and rats indicate that the glucose transport capacity of the small intestine is typically underutilized because of a fraction of time that transporters are not in contact with digesta. PMID- 10362647 TI - Functional characteristics of basolateral peptide transporter in the human intestinal cell line Caco-2. AB - The apical H+-coupled peptide transporter (PEPT1) and basolateral peptide transporter in human intestinal Caco-2 cells were functionally compared by the characterization of [14C]glycylsarcosine transport. The glycylsarcosine uptake via the basolateral peptide transporter was less sensitive to medium pH than uptake via PEPT1 and was not transported against the concentration gradient. Kinetic analysis indicated that glycylsarcosine uptake across the basolateral membranes was apparently mediated by a single peptide transporter. Small peptides and beta-lactam antibiotics inhibited glycylsarcosine uptake by the basolateral peptide transporter, and these inhibitions were revealed to be competitive. Comparison of the inhibition constant values of various beta-lactam antibiotics between PEPT1 and the basolateral peptide transporter suggested that the former had a higher affinity than the latter. A histidine residue modifier, diethyl pyrocarbonate, inhibited glycylsarcosine uptake by both transporters, although the inhibitory effect was greater on PEPT1. These findings suggest that a single facilitative peptide transporter is expressed at the basolateral membranes of Caco-2 cells and that PEPT1 and the basolateral peptide transporter cooperate in the efficient transepithelial transport of small peptides and peptidelike drugs. PMID- 10362649 TI - Rat intestinal alpha1-antitrypsin secretion is regulated by triacylglycerol feeding. AB - alpha1-Antitrypsin (AAT) is secreted by the enterocyte, but its regulation of expression, intramucosal distribution, and functional status are unclear. After corn oil gavage (plus Pluronic L-81 to block chylomicron release), rat intestine was examined for mRNA encoding AAT, immunoreactivity by light and electron microscopy, and protein content by Western blot. Species-specific antisera used were raised against both AAT and surfactant-like particle (SLP), a membrane secreted by the enterocyte in response to fat feeding. Purified luminal SLP was fractionated by Bio-Gel P-200 chromatography to assess its interaction with AAT. Triacylglycerol feeding maximally increased mucosal mRNA-encoding AAT and AAT intracellular protein content by 3 and 5 h, respectively. Immunocytochemistry revealed predominance of AAT in basolateral spaces around enterocytes and Pluronic-blocked extracellular accumulation of AAT, patterns nearly identical to those of secreted SLP. About 10% of AAT was reversibly associated with SLP. Luminal AAT was smaller (51 kDa) than mature AAT (55 kDa) and did not form a complex with pancreatic elastase. When the common bile duct was tied, excluding pancreatic proteases from the lumen, mature AAT that was cleaved by pancreatic elastase was secreted. The luminal secretion of AAT and its reversible association with SLP suggest an intracellular association and a possible role for AAT during lipid digestion and absorption. PMID- 10362650 TI - IL-2-deficient mice raised under germfree conditions develop delayed mild focal intestinal inflammation. AB - Interleukin-2 (IL-2) amplifies immune stimuli and influences B cell differentiation. IL-2-deficient mice spontaneously develop intestinal inflammation if raised under specific pathogen-free (SPF) conditions. We quantitatively determined the aggressiveness and kinetics of gastrointestinal and hepatic inflammation in the presence or absence of viable bacteria in IL-2 deficient mice. Breeding colonies were maintained under SPF and germfree (GF) conditions. Intestinal tissues, serum, and mesenteric lymph nodes were obtained from mice at different ages for blind histological scoring, immunoglobulin measurements, mucosal T cell infiltration, and cytokine secretion. GF IL-2 -/- mice developed mild, focal, and nonlethal intestinal inflammation with delayed onset, whereas the more aggressive inflammation in SPF IL-2 -/- mice led to their death between 28 and 32 wk. Periportal hepatic inflammation was equal in the presence or absence of bacterial colonization. Intestinal immunoglobulin secretion decreased significantly by 13 wk of age in IL-2 -/- mice in both GF and SPF environments. In contrast to other genetically engineered rodents, IL-2 -/- mice develop mild focal gastrointestinal and active portal tract inflammation in the absence of viable bacteria. PMID- 10362651 TI - Effects of substance P on human colonic mucosa in vitro. AB - Previous studies indicated that the peptide substance P (SP) causes Cl--dependent secretion in animal colonic mucosa. We investigated the effects of SP in human colonic mucosa mounted in Ussing chamber. Drugs for pharmacological characterization of SP-induced responses were applied 30 min before SP. Serosal, but not luminal, administration of SP (10(-8) to 10(-6) M) induced a rapid, monophasic concentration and Cl--dependent, bumetanide-sensitive short-circuit current (Isc) increase, which was inhibited by the SP neurokinin 1 (NK1)-receptor antagonist CP-96345, the neuronal blocker TTX, the mast cell stabilizer lodoxamide, the histamine 1-receptor antagonist pyrilamine, and the PG synthesis inhibitor indomethacin. SP caused TTX- and lodoxamide-sensitive histamine release from colonic mucosa. Two-photon microscopy revealed NK1 (SP)-receptor immunoreactivity on nerve cells. The tyrosine kinase inhibitor genistein concentration dependently blocked SP-induced Isc increase without impairing forskolin- and carbachol-mediated Isc increase. We conclude that SP stimulates Cl -dependent secretion in human colon by a pathway(s) involving mucosal nerves, mast cells, and the mast cell product histamine. Our results also indicate that tyrosine kinases may be involved in this SP-induced response. PMID- 10362652 TI - Carbachol activates ERK2 in isolated gastric parietal cells via multiple signaling pathways. AB - We previously reported that both carbachol and epidermal growth factor (EGF) are potent inducers of the extracellular signal-regulated protein kinases (ERKs) in isolated gastric canine parietal cells and that induction of these kinases leads to acute inhibitory and chronic stimulatory effects on gastric acid secretion. In this study we investigated the molecular mechanisms responsible for these effects. Both carbachol (100 microM) and EGF (10 nM) induced Ras activation. The role of Ras in ERK2 induction was examined by transfecting parietal cells with a vector expressing hemoagglutinin (HA)-tagged ERK2 (HA-ERK2) together with a dominantly expressed mutant (inactive) ras gene. HA-ERK2 activity was quantitated by in-gel kinase assays. Dominant negative Ras reduced carbachol induction of HA ERK2 activity by 60% and completely inhibited the stimulatory effect of EGF. Since Ras activation requires the assembly of a multiprotein complex, we examined the effect of carbachol and EGF on tyrosyl phosphorylation of Shc and its association with Grb2 and the guanine nucleotide exchange factor Sos. Western blot analysis of anti-Shc immunoprecipitates with an anti-phosphotyrosine antibody demonstrated that both carbachol and EGF induced tyrosyl phosphorylation of a major 52-kDa shc isoform. Grb2 association with Shc was demonstrated by blotting Grb2 immunoprecipitates with an anti-Shc antibody. Probing of anti-Sos immunoprecipitates with an anti-Grb2 antibody revealed that Sos was constitutively bound to Grb2. To examine the functional role of Sos in ERK2 activation, we transfected parietal cells with the HA-ERK2 vector together with a dominantly expressed mutant (inactive) sos gene. Dominant negative Sos did not affect carbachol stimulation of HA-ERK2 but inhibited the stimulatory effect of EGF by 60%. We then investigated the role of betagamma-subunits in carbachol induction of HA-ERK2. Parietal cells were transfected with the HA-ERK2 vector together with a vector expressing the carboxy terminus of the beta-adrenergic receptor kinase 1, known to block signaling mediated by betagamma-subunits. In the presence of this vector, carbachol induction of HA-ERK2 was inhibited by 40%. Together these data suggest that, in the gastric parietal cells, carbachol activates the ERKs through Ras- and betagamma-dependent mechanisms that require guanine nucleotide exchange factors other than Sos. PMID- 10362653 TI - MRP3, a new ATP-binding cassette protein localized to the canalicular domain of the hepatocyte. AB - Bile secretion in liver is driven in large part by ATP-binding cassette (ABC) type proteins that reside in the canalicular membrane and effect ATP-dependent transport of bile acids, phospholipids, and non-bile acid organic anions. Canalicular ABC-type proteins can be classified into two subfamilies based on membrane topology and sequence identity: MDR1, MDR3, and SPGP resemble the multidrug resistance (MDR) P-glycoprotein, whereas MRP2 is similar in structure and sequence to the multidrug resistance protein MRP1 and transports similar substrates. We now report the isolation of the rMRP3 gene from rat liver, which codes for a protein 1522 amino acids in length that exhibits extensive sequence similarity with MRP1 and MRP2. Northern blot analyses indicate that rMRP3 is expressed in lung and intestine of Sprague-Dawley rats as well as in liver of Eisai hyperbilirubinemic rats and TR- mutant rats, which are deficient in MRP2 expression. rMRP3 expression is also transiently induced in liver shortly after birth and during obstructive cholestasis. Antibodies raised against MRP3 recognize a polypeptide of 190-200 kDa, which is reduced in size to 155-165 kDa after treatment with endoglycosidases. Immunoblot analysis and immunoconfocal microscopy indicate that rMRP3 is present in the canalicular membrane, suggesting that it may play a role in bile formation. PMID- 10362654 TI - Duodenal acid-induced gastric relaxation is mediated by multiple pathways. AB - In this study, we used an in vivo anesthetized rat model to investigate the mechanisms responsible for duodenal acid-induced inhibition of gastric motility. Intraduodenal infusion of HCl produced a rate-dependent decrease in intragastric pressure. Infusion of HCl at 2 ml/h produced a physiological plasma secretin level and elicited a decrease in intragastric pressure of 3.0 +/- 0. 2 cmH20. Infusion of rabbit secretin antiserum reduced the acid-induced inhibition of gastric motility by 85 +/- 5%, suggesting mediation mainly by endogenous secretin. Administration of the cholecystokinin (CCK)-A antagonist MK-329 caused only a modest 10 +/- 3% reduction in gastric relaxation, whereas the serotonin antagonist ICS-205930 had no effect. In contrast, immunoneutralization with the secretin antibody caused only a 15% reduction in the relaxation evoked by a higher rate of HCl infusion (3 ml/h), whereas MK-329 and ICS-205930 caused a 20 +/- 4% reduction and no reduction, respectively. Bilateral truncal vagotomy or perivagal application of capsaicin completely abolished gastric relaxation in response to low rates (1-2 ml/h) of 0.1 N HCl infusion but only partially affected gastric relaxation in response to a higher infusion rate (3 ml/h). These observations indicate that multiple pathways mediate the duodenal acid-induced inhibition of gastric motility. At low rates of HCl infusion, gastric relaxation is mediated primarily by endogenous secretin, which acts through vagal afferent pathways. At higher rates of HCl infusion, gastric relaxation is mediated by endogenous secretin, CCK, and possibly by the direct action of HCl on vagal afferent pathways or yet unidentified neuropathways. PMID- 10362655 TI - Effect of diaspirin cross-linked hemoglobin on normal and postischemic microcirculation of the rat pancreas. AB - Microcirculatory alterations with reduced nutritive supply to the pancreas could be the cause of hyperamylasemia, which occurs in some patients receiving the vasoactive oxygen carrier diaspirin cross-linked hemoglobin (DCLHb) in clinical studies. Therefore, the effects of DCLHb on rat pancreas microcirculation were evaluated. Anesthetized Sprague-Dawley rats received one of the following treatments during baseline conditions (n = 7 rats/group): 10% hydroxyethyl starch (HAES) (0.4 ml/kg), DCLHb (400 mg/kg), or DCLHb (1,400 mg/kg). After 1 h of complete, reversible pancreatic ischemia, other animals received 10% HAES (0.4 ml/kg) or DCLHb (400 mg/kg) during the onset of reperfusion. The number of red blood cell-perfused capillaries (functional capillary density, FCD) and the level of leukocyte adherence in postcapillary venules in the pancreas were assessed by means of intravital microscopy during 2 h after treatment. In the nonischemic groups, FCD was 18% greater after DCLHb (1,400 mg/kg) than after 10% HAES treatment without any increase in leukocyte adherence. In the inschemia reperfusion (I/R) 10% HAES group, FCD was significantly (P < 0.05) lowered, leukocyte adherence enhanced, and mean arterial pressure (MAP) reduced by 31% compared with nonischemic animals. DCLHb treatment in the I/R group resulted in a slight increase in FCD, a significant (P < 0.05) reduction of leukocyte adherence, and a complete restoration of MAP compared with the animals of the I/R control group. Thus our data provide no evidence for a detrimental effect on the pancreatic microcirculation or an enhanced risk of postischemic pancreatitis by DCLHb. PMID- 10362656 TI - PO2 measurements in the rat intestinal microcirculation. AB - Microvascular partial oxygen pressure (PO2) data measured with the quenched phosphorescence of palladium-porphyrin (Pd-porphyrin) with the use of optical fibers have provided new insight into the behavior of the microvascular oxygenation in models of shock. However, the actual microcirculatory compartment measured by this fiber technique has not yet been demonstrated. The purpose of this study was to investigate whether the PO2 of the intestines, as measured using a fiber phosphorometer, reflects the microvascular compartment. To this end, a new intravital phosphorometer with an improved sensitivity to high-PO2 levels (up to 180 mmHg) was developed and validated. With this setup, PO2 values were measured at different inspired oxygen fractions (15, 25, and 50% oxygen) in first-order arterioles, capillaries, and venules of the ileum of rats. Simultaneously, the PO2 was measured with an optical fiber attached to another phosphorometer. PO2 measurements with the fiber phosphorometer show excellent correlation with the PO2 in the capillaries and first-order venule vessels (r2 = 0.94, slope 0.99). We therefore conclude that values measured with the fiber phosphorometer correlate with the capillary and venular PO2. PMID- 10362658 TI - Cardiac myofibrillar and sarcoplasmic reticulum function are not depressed in insulin-resistant JCR:LA-cp rats. AB - Depressed myofibrillar Ca2+-ATPase activity and sarcoplasmic reticulum (SR) Ca2+ uptake are important mechanisms that are responsible for the cardiac dysfunction exhibited by insulin-deficient (type I) diabetic animals. The JCR:LA-cp rat is a model for type II non-insulin-dependent diabetes mellitus (NIDDM). This rat is insulin resistant, obese, and has high levels of circulating glucose, cholesterol, insulin, and triglycerides. The purpose of this study was to determine whether changes in cardiac myofibrillar, SR, and cardiomyocyte function exist in this model of type II diabetes. Myofibrils and SR were isolated from hearts by differential centrifugation. Surprisingly, we found that myofibrillar Ca2+-ATPase activities were unaltered in these animals. Ca2+ uptake in isolated SR fractions was increased in diabetic cp/cp rats, whereas Ca2+-ATPase activity and ryanodine binding were unchanged. Cardiomyocytes isolated from hearts of control and experimental animals had similar active cell shortening and intracellular Ca2+ concentration under basal conditions and in response to caffeine. Our data argue against the presence of a cardiomyopathy in this diabetic model and suggest that insulin may be an important factor in the cardiomyopathy observed in type I diabetic models. PMID- 10362657 TI - NO releases bombesin-like immunoreactivity from enteric synaptosomes by cross activation of protein kinase A. AB - The effect of nitric oxide (NO) on the release of bombesin-like immunoreactivity (BLI) was examined in synaptosomes of rat small intestine. The NO donor S-nitroso N-acetylpenicillamine (SNAP; 10(-7) to 10(-4) M) significantly stimulated BLI release. In the presence of the NO scavenger oxyhemoglobin (10(-3) M) or the guanylate cyclase inhibitor ODQ (10(-5) M), SNAP-induced BLI release was antagonized. In addition, SNAP increased the synaptosomal cGMP content and elevation of cGMP levels by zaprinast (3 x 10(-5) M), an inhibitor of the cGMP specific phosphodiesterase (PDE) type 5, and increased basal and SNAP-induced BLI release. NO-induced BLI release was blocked by Rp-adenosine 3',5'-cyclic monophosphorothioate (3 x 10(-5) M and 10(-4) M), an inhibitor of the cAMP dependent protein kinase A, whereas KT-5823 (3 x 10(-6) M) and Rp-8-(4 chlorophenylthio)-cGMP (5 x 10(-5) M), inhibitors of the cGMP-dependent protein kinase G, had no effect. Because cGMP inhibits the cAMP-specific PDE3, thereby increasing cAMP levels, the role of PDE3 was investigated. Trequinsin (10(-8) M), a specific blocker of PDE3, stimulated basal BLI release but had no additive effect on NO-induced release, suggesting a similar mechanism of action. These data demonstrate that because of a cross-activation of cAMP-dependent protein kinase A by endogenous cGMP BLI can be released by NO from enteric synaptosomes. PMID- 10362659 TI - Active renin and angiotensinogen in cardiac interstitial fluid after myocardial infarction. AB - The renin-angiotensin system promotes cardiac hypertrophy after myocardial infarction. The purpose of this study was to measure renin and angiotensinogen in plasma and myocardium 10 days after myocardial infarction. Infarction involving 45 +/- 4% of left ventricular circumference with accompanying hypertrophy was induced in rats (n = 14). Plasma and myocardial renin were increased after infarction compared with sham controls (n = 8) (27.4 +/- 3.2 vs. 7.5 +/- 1.8 ng ANG I. ml plasma. h-1, P < 0.0002; and 8.8 +/- 1.6 vs. 2. 5 +/- 0.1 ng ANG I. g myocardium-1. h-1, P < 0.008, respectively). After infarction, myocardial renin was correlated with infarct size (r = 0.62, P < 0.02) and plasma renin (r = 0.55, P < 0.04). Plasma angiotensinogen decreased in infarct animals, but myocardial angiotensinogen was not different from shams (1.1 +/- 0.08 vs. 2.03 +/- 0.06 nM/ml plasma, P < 0.002; and 0.081 +/- 0.008 vs. 0.070 +/- 0.004 nM/g myocardium, respectively). In conclusion, myocardial renin increased after infarction in proportion to plasma renin and infarct size, and myocardial angiotensinogen was maintained after infarction despite decreased plasma angiotensinogen and increased levels of myocardial renin. PMID- 10362660 TI - Characterization of a newly found stretch-activated KCa,ATP channel in cultured chick ventricular myocytes. AB - With the use of the patch-clamp technique, five kinds of stretch-activated (SA) ion channels were identified on the basis of their single-channel conductances and ion selectivities in cultured chick ventricular myocytes. Because a high conductance K+-selective channel predominated among these channels, we concentrated on characterizing its properties mostly using excised inside-out patches. With 145 mM KCl solution in the pipette and the bath, the channel had a conductance of 199.8 +/- 8.2 pS (n = 22). The ion selectivities among K+, Na+, Ca2+, and Cl- as estimated from their permeability ratios were PNa/PK = 0.03, PCa/PK = 0.025, and PCl/PK = 0.026. The probability of the channel being open (Po) increased with the Ca2+ concentration in the bath ([Ca2+]b; dissociation constant Kd = 0.51 microM at +30 mV) and membrane potential (voltage at half maximal Po = 39.4 mV at 0.35 microM [Ca2+]b). The channel was blocked by gadolinium, tetraethylammonium, and charybdotoxin from the extracellular surface and, consequently, was identified as a Ca2+-activated K+ (KCa) channel type. The channel was also reversibly activated by ATP applied to the intracellular surface (Kd = 0.74 mM at 0.10 microM [Ca2+]b at +30 mV). From these data taken together, we concluded that the channel is a new type of KCa channel that could be designated as an "SA KCa,ATP channel." To our knowledge, this is the first report of KCa channel in heart cells. PMID- 10362661 TI - Systemic arterial compliance is reduced in young patients with IDDM. AB - Arterial elastic properties are altered with increasing age and in various disease states, including non-insulin-dependent diabetes mellitus (NIDDM). Whether young patients with insulin-dependent diabetes mellitus (IDDM) have reduced arterial compliance before developing endothelial dysfunction or overt micro- and macrovascular disease is unclear. Systemic arterial compliance and endothelium-dependent, flow-mediated vasodilation (FMD) was assessed in 25 individuals with uncomplicated IDDM (23 +/- 4 yr, 14 females and 11 males) and compared with 30 age-matched controls (15 females and 15 males). Arterial compliance was determined via simultaneous measurements of aortic blood flow and carotid arterial pressure. The relationship between arterial compliance and endothelial function (assessed by brachial artery FMD) was also examined. Arterial compliance was 29% lower in IDDM subjects compared with control subjects (0.46 +/- 0.05 vs. 0.65 +/- 0.07 arbitrary compliance units, P < 0.05). Blood pressure, lipid levels, and daily energy expenditure (a measure of physical activity levels) were not different between groups. Compliance in the IDDM group was not related to the integrity of endothelial vasodilator function, disease duration, or degree of glycemic control. Arterial compliance is reduced in young patients with IDDM before the development of overt micro- or macrovascular disease. Early assessment of arterial compliance may be useful in predicting the development of diabetic vascular complications. PMID- 10362662 TI - Adventitia-dependent relaxations of canine basilar arteries transduced with recombinant eNOS gene. AB - We recently reported that expression of recombinant endothelial nitric oxide (NO) synthase (eNOS) gene in adventitial fibroblasts restores NO formation in canine cerebral arteries without endothelium in response to bradykinin ex vivo and in vivo. The present study was designed to further characterize the stimuli that can activate recombinant eNOS enzyme expressed in the adventitia of cerebral arteries. To stimulate recombinant eNOS, we used serum (0. 1-10%), substance P (10(-11)-3 x 10(-9) M), and ANG II (10(-7)-10(-5) M) because they increase intracellular calcium concentrations in fibroblasts. Endothelium-denuded segments of canine basilar arteries were incubated with an adenoviral vector encoding beta galactosidase gene or eNOS gene for 30 min at 37 degrees C. After 24 h, vasomotor activity and cGMP formation in eNOS or beta-galactosidase arteries were examined by isometric force recording and by radioimmunoassay, respectively. In control arteries and beta-galactosidase gene-transduced arteries, serum caused concentration-dependent contractions, whereas in recombinant eNOS gene-transduced arteries, serum produced concentration-dependent relaxations. Substance P and ANG II had no effect on vascular tone in control and beta-galactosidase arteries but caused concentration-dependent relaxations as well as a significant increase in cGMP levels in eNOS arteries. These relaxations were blocked by the NOS inhibitor NG-nitro-L-arginine methyl ester. Chemical treatment or mechanical inactivation of adventitial function significantly attenuated substance P-induced relaxations and ANG II-induced relaxations. These findings demonstrate that serum, substance P, and ANG II cause adventitia-dependent relaxations in cerebral arteries expressing the recombinant eNOS gene. This mechanism of vasodilatation may have beneficial effects in the prevention and treatment of vascular disorders characterized by the diminished bioavailability of NO, such as cerebral vasospasm. PMID- 10362663 TI - Molecular beta-adrenergic signaling abnormalities in failing rabbit hearts after infarction. AB - We studied alterations in the beta-adrenergic receptor (beta-AR) system of rabbit hearts during the development of heart failure (HF) after myocardial infarction (MI) to determine whether the molecular beta-AR abnormalities associated with human HF exist in this animal model. Rabbit HF was established 3 wk after left circumflex coronary artery (LCX) ligation by in vivo physiological measurements, and molecular beta-AR signaling was examined in tissue and cultured ventricular myocytes. We found that there was a significant global reduction in beta-AR density by approximately 50% in both ventricles of MI animals compared with sham operated control animals and that functional beta-AR coupling was significantly reduced. Importantly, as found in human HF, myocardial protein levels and activity of the beta-AR kinase (beta-ARK1) and Galphai were found to be significantly elevated in MI rabbits, suggesting that these molecules are contributing to myocardial dysfunction. Thus the myocardial beta-AR system of this rabbit model of HF shares important biochemical characteristics with human HF and therefore is an ideal laboratory model to investigate novel therapeutic targets for the treatment of HF. PMID- 10362664 TI - Peroxynitrite contributes to spontaneous loss of cardiac efficiency in isolated working rat hearts. AB - We examined the mechanism of the time- and protein synthesis-dependent decline in cardiac mechanical function in isolated working rat hearts. Hearts were perfused with Krebs-Henseleit buffer for 120 min in the presence or absence of the protein synthesis inhibitor cycloheximide (CX; 10 microM). Cardiac work remained stable for 60 min and then spontaneously decreased during 60-120 min of perfusion. This was accompanied by an increase in myocardial inducible nitric oxide synthase (iNOS) and xanthine oxidase (XO) activities and enhanced dityrosine formation in the perfusate, an indicator of peroxynitrite generation. CX markedly attenuated the loss in contractile function and prevented the increase in iNOS and XO activities and dityrosine level. Despite the decline in cardiac work in control hearts, the coupling between tricarboxylic acid (TCA) cycle activity and oxygen consumption remained constant in both groups. ATP, creatine phosphate, and glycogen levels were not different between control and CX groups and did not differ over 120 min of perfusion. We concluded that the delayed and spontaneous loss in myocardial mechanical function in isolated working rat hearts is 1) attenuated by CX treatment, 2) accompanied by a concomitant increase in both iNOS and XO activities and peroxynitrite generation in the heart, and 3) not dependent on a direct impairment in myocardial ATP production, myocardial oxygen consumption, or TCA cycle acetyl-CoA production but may be due to an inefficiency of the heart to utilize ATP for contractile work. PMID- 10362665 TI - Cardioprotective effects of KB-R7943: a novel inhibitor of the reverse mode of Na+/Ca2+ exchanger. AB - The novel inhibitor of the reverse mode of the Na+/Ca2+ exchanger (NCE) KB-R7943 (KB) was tested in isolated rat cardiomyocytes exposed to 80 min of simulated ischemia [substrate-free anoxia, extracellular pH (pHo) of 6.4] and 15 min of reoxygenation (pHo 7.4). At pHo 6.4, 20 micromol/l KB was required for complete inhibition of the reverse mode of NCE. Treatment with 20 micromol/l KB only during anoxia did not influence the onset of rigor contracture and intracellular pH (pHi) (monitored with 2', 7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein) but significantly reduced the cytosolic accumulation of Ca2+ (monitored with fura 2) and Na+ (monitored with sodium-binding benzofuran isophthalate). During reoxygenation, cardiomyocytes developed hypercontracture. This was significantly reduced by anoxic KB treatment. To investigate this protection against reoxygenation-induced injury in the whole heart, we exposed Langendorff-perfused rat hearts to 110 min of anoxia (pHo 6.4) and 50 min of reoxygenation (pHo 7.4). Application of 20 micromol/l KB during anoxia significantly reduced the reoxygenation-induced enzyme release. We conclude that KB offers significant protection of cardiomyocytes against Ca2+ and Na+ overload during anoxia and hypercontracture or enzyme release on reoxygenation. PMID- 10362666 TI - 5-(N-ethylcarboxamido)adenosine desensitizes the A2b-adenosine receptor in lung circulation. AB - The adenosine agonist 5-(N-ethylcarboxamido)adenosine (NECA) induces vasodilation in the pulmonary circulation via A2-adenosine-receptor activation. We addressed whether prolonged treatment with NECA desensitizes in A2-adenosine- receptor function in isolated lung and pulmonary artery smooth muscle cells (PASMC). In lung microcirculation preconstricted with a hypoxic gas, initial administration of NECA caused a 57% vasodilatory response after 3-4 min. Readministration of NECA after 45 min resulted in minimal vasodilation. The highest accumulation of PASMC cAMP occurred 3-5 min after NECA, coincident with NECA-induced vasodilation. In PASMCs treated with NECA for 45 min, cAMP did not increase. Isoproterenol- and indolidan-induced vasodilation remained intact in NECA desensitized lungs. In NECA-desensitized PASMCs, isoproterenol-induced cAMP accumulation was decreased, suggesting a common mechanism of desensitization. cAMP accumulation was decreased in cholera toxin-treated NECA-desensitized PASMCs compared with cholera toxin-treated control PASMCs, demonstrating that Gsalpha adenylyl cyclase signaling contributes to desensitization. The A2a-adenosine receptor agonist CGS-21680C neither increased cAMP accumulation in PASMCs nor attenuated NECA-induced vasodilation. These data support that the A2b-adenosine receptor is responsible for pulmonary vasodilation and desensitization through mechanisms(s) involving Gsalpha-adenylyl cyclase signaling. PMID- 10362667 TI - Activation of visceral afferents by bradykinin and ischemia: independent roles of PKC and prostaglandins. AB - We have shown that the cyclooxygenase (COX) and protein kinase C (PKC) systems both contribute to afferent activation in response to bradykinin (BK) and abdominal ischemia. Because the contribution from PKC to C fiber activation may depend, in part, on prostaglandin production, we hypothesized that an intact COX system is required for PKC-induced activation of ischemically sensitive abdominal visceral afferents by BK and abdominal ischemia. Single-unit activity of abdominal visceral C fibers was recorded from the right thoracic sympathetic chain of anesthetized cats. Three repeated injections of BK (1-2 micrograms/kg ia) produced similar increases in afferent activity from the baseline of 1.32 +/- 0.24, 1.37 +/- 0.32, and 1.41 +/- 0.24 impulses/s (n = 5). In another group of animals (n = 5), the second and third BK injections were performed after COX inhibition (indomethacin; 5 mg/kg iv) and then combined COX + PKC inhibition [PKC (19-36), 20 micrograms/kg iv], respectively. Inhibition of COX reduced (P < 0.05) the afferent response to BK (0.59 +/- 0.12 impulses/s) compared with the unblocked condition (1.14 +/- 0.27 impulses/s), whereas combined COX + PKC inhibition further attenuated the increase from baseline (0.18 +/- 0.09 impulses/s; P < 0.05). Similar results were obtained in a third group of cats when the antagonists were administered in reverse order (n = 7). In a fourth group of cats (n = 9) that were pretreated with indomethacin, ischemia increased afferent activity (0.78 +/- 0.17 impulses/s). However, neural activity was attenuated (0.51 +/- 0.14 impulses/s; P < 0.05) during a second bout of ischemia in the presence of indomethacin + PKC-(19-36). These results suggest that the contribution from PKC to the activation of ischemically sensitive C fibers, particularly by BK, does not require an intact cyclooxygenase system. PMID- 10362668 TI - ATP reduces macromolecule permeability of endothelial monolayers despite increasing [Ca2+]i. AB - We investigated the relationship between the ATP-evoked rise of cytosolic Ca2+ concentration ([Ca2+]i) and barrier function in porcine aortic endothelial monolayers. ATP (0.01-100 microM) induced a transient rise of [Ca2+]i and reduced permeability in a concentration-dependent manner. In contrast, the Ca2+ ionophore ionomycin (1 microM) elicited a rise in [Ca2+]i comparable to that induced by ATP (10 microM), but it increased permeability. For the reduction of permeability, nucleotides were found to be in the following order of potency: ATP = ATPgammaS > ADP = UTP. Blockade of adenosine receptors by 8-phenyltheophylline (10 microM) did not affect ATP (10 microM)-induced reduction of permeability. ATP reduced permeability even in endothelial monolayers that had been loaded with the Ca2+ chelator BAPTA to prevent the rise in [Ca2+]i. U-73122 (1 microM), an inhibitor of phospholipase C (PLC), completely abolished the effect of ATP (10 microM) on permeability. It also abolished the translocation of protein kinase C (PKC) in response to ATP, which could also be achieved by the PKC inhibitors Go-6976 (100 nM) or bisindolylmaleimide I (1 microM). In the presence of PKC inhibitors, however, the permeability effect of ATP was not affected. The presence of inhibitors of adenylate or guanylate cyclase (50 microM SQ-22536 or 20 microM ODQ) prevented changes in cyclic nucleotides but did not affect the permeability effects of ATP. The study shows that ATP reduces macromolecule permeability via a PLC-mediated mechanism that is independent of the concomitant effects of ATP on cytosolic Ca2+, cyclic nucleotides, or PKC. PMID- 10362670 TI - Further evidence for the selective disruption of intercellular communication by heptanol. AB - The lack of selective gap junctional uncoupling agents has hampered evaluation of the contribution of intercellular communication to pharmacomechanical coupling and vascular contractility. Thus we further explored the utility and selectivity of heptanol as a gap junctional uncoupling agent in isolated rat aortic rings. Fifty-two aortic rings were obtained from 15 rats and were precontracted to approximately 75% of maximum with phenylephrine (PE). When contraction achieved steady state (approximately 5 min), a single concentration of heptanol (200 microM) was added to each aortic ring at 1- to 3-min intervals for up to 42 min post-PE addition. At early time points (5-10 min after PE), heptanol elicited an approximately 50% loss of tension (i.e., relaxation). At subsequent time points post-PE, a gradual and time-dependent decrease in the magnitude of the heptanol induced relaxation was observed until, after approximately 40 min, addition of heptanol was associated with little, if any, detectable relaxation. Linear regression analysis of the magnitude of the heptanol-induced relaxation vs. the square root of the elapsed time interval (from addition of PE) revealed a highly significant negative correlation (P < 0.001, R = 0.81). Studies conducted on KCl precontracted aortic rings revealed no detectable heptanol-induced relaxation after development of the steady-state KCl-induced contraction. These data extend our previous observations to further document the potential utility of heptanol as a "relatively selective" uncoupling agent. PMID- 10362669 TI - Activation of GABAA but not GABAB receptors in the NTSblocked bradycardia of chemoreflex in awake rats. AB - In the present study we analyzed effects of bilateral microinjections of muscimol (a GABAA agonist) and baclofen (a GABAB agonist) into the nucleus tractus solitarius (NTS) on bradycardic and pressor responses to chemoreflex activation (potassium cyanide, 40 micrograms/rat iv) in awake rats. Bilateral microinjections of muscimol (25 and 50 pmol/50 nl) into the NTS increased baseline mean arterial pressure (MAP): 119 +/- 8 vs. 107 +/- 2 mmHg (n = 6) and 121 +/- 8 vs. 103 +/- 3 mmHg (n = 6), respectively. Muscimol at 25 pmol/50 nl reduced the bradycardic response to chemoreflex activation 5 min after microinjection; with 50 pmol/50 nl the bradycardic response to chemoreflex activation was reduced 5, 15, 30, and 60 min after microinjection. Neither muscimol dose produced an effect on the pressor response of the chemoreflex. Effects of muscimol (50 pmol/50 nl) on basal MAP and on the bradycardic response of the chemoreflex were prevented by prior microinjection of bicuculline (a GABAA antagonist, 40 pmol/50 nl) into the NTS. Bilateral microinjections of baclofen (12.5 and 25 pmol/50 nl) into the NTS produced an increase in baseline MAP [137 +/- 9 vs. 108 +/- 4 (n = 7) and 145 +/- 5 vs. 105 +/- 2 mmHg (n = 7), respectively], no changes in basal heart rate, and no effects on the bradycardic response; 25 pmol/50 nl only attenuated the pressor response to chemoreflex activation. The data show that activation of GABAA receptors in the NTS produces a significant reduction in the bradycardic response, whereas activation of GABAB receptors produces a significant reduction in the pressor response of the chemoreflex. We conclude that 1) GABAA but not GABAB plays an inhibitory role in neurons of the lateral commissural NTS involved in the parasympathetic component of the chemoreflex and 2) attenuation of the pressor response of the chemoreflex by activation of GABAB receptors may be due to inhibition of sympathoexcitatory neurons in the NTS or may be secondary to the large increase in baseline MAP produced by baclofen. PMID- 10362671 TI - Role of central AT1 and V1 receptors in cardiovascular adaptation to hemorrhage in SD and renin TGR rats. AB - In acute experiments, intracranially applied angiotensin II and vasopressin elicit significant cardiovascular effects. The purpose of the present study was to find out whether chronic intrabrain elevation of these peptides, occurring in the renin transgenic TGR(mRen2)27 (TGR) rats, results in an alteration of the cardiovascular control. Mean arterial blood pressure (MAP) and heart rate responses to hypovolemia were examined in hypertensive TGR and normotensive Sprague-Dawley (SD) rats under control conditions and during blockade of central AT1 or V1 receptors. Both groups received cerebroventricular infusions of either 1) cerebrospinal fluid (series 1), 2) AT1 receptors antagonist (AT1ANT, series 2), or 3) V1 receptors antagonist (V1ANT, series 3). Blockade of AT1 and V1 receptors decreased MAP in TGR but not in SD rats. In SD rats, bleeding elicited a similar decrease of MAP in each series and a transient increase of heart rate in series 3. In TGR, hemorrhage caused bradycardia and decrease of MAP, which was greater than in SD rats. Hemorrhagic hypotension in TGR was abolished by V1ANT and bradycardia by V1ANT or AT1ANT. The results demonstrate remarkable differences in cardiovascular adjustment to hemorrhage in SD and TGR rats and provide evidence for enhanced involvement of central V1 and AT1 receptors in the regulation of blood pressure during hypovolemia in TGR. Central V1 vasopressin receptors play a crucial role in eliciting posthemorrhagic hypotension and bradycardia in this strain. PMID- 10362672 TI - Activation of Akt/protein kinase B after stimulation with angiotensin II in vascular smooth muscle cells. AB - Involvement of Akt/Protein kinase B (PKB), a serine/threonine kinase with a pleckstrin-homology domain, in angiotensin II (ANG II)-induced signal transduction was investigated in cultured vascular smooth muscle cells (VSMC). Stimulation of the cells with ANG II led to a marked increase in the kinase activity of Akt/PKB, which coincided with Ser-473 phosphorylation. ANG II stimulated Akt/PKB activation was rapid, concentration dependent, and inhibited by the AT1-receptor antagonist CV-11974, but not by pertussis toxin. Akt/PKB activity was stimulated by the Ca2+ ionophore ionomycin, suggesting the possible involvement of Ca2+ in ANG II-stimulated Akt/PKB activation. However, blockade of Ca2+ mobilization by BAPTA-AM only partially inhibited ANG II-stimulated Akt/PKB activation. ANG II-stimulated Akt/PKB activation was inhibited by the tyrosine kinase inhibitors genistein and herbimycin A and by the phosphatidylinositol 3 kinase (PI3K) inhibitors wortmannin and LY-294002. These results indicate that ANG II stimulates Akt/PKB activity via AT1 receptors in VSMC and that the activities of tyrosine kinase and PI3K are required for this activation. PMID- 10362673 TI - Vasorelaxing effects of atrial and brain natriuretic peptides on coronary circulation in heart failure. AB - Natriuretic peptide (NP) receptor has been postulated to be downregulated under a high concentration of atrial NP (ANP) in congestive heart failure (CHF), but limited information is available on how the vascular functional responsiveness to NPs is altered in coronary circulation during CHF. We assessed the relaxant effects of ANP, brain NP (BNP), and other vasodilators in isolated coronary arteries obtained from dogs with and without severe CHF induced by rapid right ventricular pacing. In CHF dogs, plasma ANP and cGMP concentrations were elevated compared with control dogs. In CHF arteries the relaxant effects of ANP and BNP (10(-8) and 10(-7) mol/l) were suppressed compared with control arteries. Nitroglycerin, nitric oxide, 8-bromo-cGMP, and beraprost sodium produced similar concentration-response curves in both arteries. The addition of 10(-7) mol/l ANP increased the level of tissue cGMP in control arteries, but not in CHF arteries. We conclude that there was a specific reduction in the relaxant effects of ANP and BNP in isolated coronary arteries in severe CHF dogs, which suggests the possibility of the downregulation of NP receptors coupled to guanylate cyclase. PMID- 10362674 TI - Leukocyte-endothelial cell interactions in nitric oxide synthase-deficient mice. AB - Nitric oxide (NO) is known to be an important endogenous modulator of leukocyte endothelial cell interactions within the microcirculation. We examined leukocyte rolling and adhesion under baseline conditions and following thrombin (0.25 U/ml) superfusion in the mesentery of wild-type, inducible NOS (iNOS)-deficient (-/-), neuronal NOS (nNOS) -/-, and endothelial cell NOS (ecNOS) -/- mice to further our understanding of NO and leukocyte function. Baseline leukocyte rolling (cells/min) was significantly elevated in both the nNOS -/- (30.0 +/- 4.0) and ecNOS -/- mice (67.0 +/- 12.0) compared with wild-type mice (11.0 +/- 1.4). In addition, baseline leukocyte adherence (cells/100 micrometers of vessel) was also significantly elevated in the nNOS -/- (5.2 +/- 1.0) and ecNOS -/- (13.0 +/- 1.3) compared with wild-type animals (1.3 +/- 0.5). Deficiency of iNOS had no effect on baseline leukocyte rolling or adhesion in the mesentery. Baseline surface expression of P-selectin was observed in 68.0 +/- 9.0% of intestinal venules in ecNOS -/- mice compared with 10.0 +/- 2.0% in wild-type mice. Additional studies demonstrated that administration of an anti-P-selectin monoclonal antibody (RB40. 34) or the soluble P-selectin ligand, PSGL-1, completely inhibited the increased rolling and firm adhesion response in nNOS -/- and ecNOS -/- mice. Transmigration of neutrophils into the peritoneum following thioglycollate injection was also significantly augmented in nNOS -/- and ecNOS -/- mice. These studies clearly indicate the NO derived from both nNOS and ecNOS is critical in the regulation of leukocyte-endothelial cell interactions. PMID- 10362675 TI - Nitric oxide in the regulation of vasomotor tone in human skeletal muscle. AB - The role of nitric oxide (NO) as a regulator of vasomotor tone has been investigated in resting and exercising human skeletal muscle. At rest, NO synthase (NOS) inhibition by intra-arterial infusion of NG-monomethyl-L-arginine decreased femoral artery blood flow (FABF, ultrasound Doppler) from 0.39 +/- 0.08 to 0.18 +/- 0.03 l/min (P < 0. 01), i.e., by approximately 52%, and increased leg O2 extraction from 62.1 +/- 9.8 to 100.9 +/- 4.5 ml/l (P < 0.004); thus leg O2 uptake (VO2, 22 +/- 4 ml/min, approximately 0.75 ml. min-1. 100 g-1) was unaltered [not significant (P = NS)]. Mean arterial pressure (MAP) increased by 8 +/- 2 mmHg (P < 0.01). Heart rate (HR, 53 +/- 3 beats/min) was unaltered (P = NS). The NOS inhibition had, however, no effect on the initial rate of rise or the magnitude of FABF (4.8 +/- 0.4 l/min, approximately 163 ml. min-1. 100 g-1), MAP (117 +/- 3 mmHg), HR (98 +/- 5 beats/min), or leg VO2 (704 +/- 55 ml/min, approximately 24 ml. min-1. 100 g-1, P = NS) during submaximal, one-legged, dynamic knee-extensor exercise. Similarly, FABF (7.6 +/- 1.0 l/min, approximately 258 ml. min-1. 100 g-1), MAP (140 +/- 8 mmHg), and leg VO2 (1,173 +/- 139 ml/min, approximately 40 ml. min-1. 100 g-1) were unaffected at termination of peak effort (P = NS). Peak HR (137 +/- 3 beats/min) was, however, lowered by 10% (P < 0.01). During recovery, NOS inhibition reduced FABF by approximately 34% (P < 0.04), which was compensated for by an increase in the leg O2 extraction by approximately 41% (P < 0.04); thus leg VO2 was unaltered (P = NS). In conclusion, these findings indicate that NO is not essential for the initiation or maintenance of active hyperemia in human skeletal muscle but support a role for NO during rest, including recovery from exercise. Moreover, changes in blood flow during rest and recovery caused by NOS inhibition are accompanied by reciprocal changes in O2 extraction, and thus VO2 is maintained. PMID- 10362676 TI - Heat shock protein expression protects against cerebral ischemia and monoamine overload in rat heatstroke. AB - This study attempted to ascertain whether the ischemic damage to neurons and monoamine overload in brain that occur during rat heatstroke can be attenuated by heat shock protein (HSP) 72 induction. Effects of heatstroke on mean arterial pressure (MAP), cerebral blood flow (CBF), brain dopamine (DA) and serotonin (5 HT) release, and neural damage score were assayed in rats 0, 16, or 48 h after heat shock (42 degrees C for 15 min) or chemical stress (5 mg/kg sodium arsenite ip). Brain HSP 72 in rats after heat shock or chemical stress was detected by Western blot, and brain monoamine was determined by a microdialysis probe combined with high-performance liquid chromatography. Heatstroke was induced by exposing the animal to a high ambient temperature (43 degrees C); the moment at which MAP and CBF decreased from their peak values was taken as the time of heatstroke onset. Prior heat shock or chemical stress conferred significant protection against heatstroke-induced hyperthermia, arterial hypotension, cerebral ischemia, cerebral DA and 5-HT overload, and neural damage and correlated with expression of HSP 72 in brain at 16 h. However, at 48 h, when HSP 72 expression returned to basal values, the above responses that occurred during the onset of heatstroke were indistinguishable between the two groups (0 h vs. 48 h). These results lead to the hypothesis that the brain can be preconditioned by thermal or chemical injury, that this preconditioning will induce HSP 72, and that HSP 72 induction will correlate quite well with anatomic, histochemical, and hemodynamic protection in rat heatstroke. PMID- 10362677 TI - Angiotensin II and mechanical stretch induce production of tumor necrosis factor in cardiac fibroblasts. AB - To determine whether ANG II as well as mechanical stress affect the production of tumor necrosis factor (TNF) in the heart, neonatal rat cardiac myocytes and fibroblasts were cultured separately and treated for 6 h with ANG II, lipopolysaccharide (LPS), or cyclic mechanical stretch. LPS induced the production of TNF in cardiac myocytes and fibroblasts. However, TNF synthesis in fibroblasts was 20- to 40-fold higher than in myocytes. ANG II (>/=10(-8) M) and mechanical stretch stimulated the production of TNF in cardiac fibroblasts but not in myocytes. Furthermore, both ANG II and LPS increased the expression of TNF alpha mRNA in cardiac fibroblasts. Isoproterenol inhibited both LPS- and ANG II induced production of TNF in cardiac fibroblasts with increasing intracellular cAMP level. Moreover, both isoproterenol and dibutyryl cAMP inhibited LPS-induced TNF-alpha mRNA expression. Thus activation of the renin-angiotensin system, as well as mechanical stress, can stimulate production of TNF in cardiac fibroblasts. Furthermore, beta-adrenergic receptors may be responsible for the regulation of TNF synthesis at the transcriptional level by elevating intracellular cAMP. PMID- 10362678 TI - Characterization of natriuretic peptide production by adult heart atria. AB - The cardiac polypeptide hormones atrial natriuretic factor (ANF) and brain natriuretic peptide (BNP) are synthesized and costored by atrial cardiocytes and share receptors and many biologic properties. Although some aspects of their synthesis and release are specific for each peptide, it is not clear whether they share intracellular sorting and secretory mechanisms. In the present work we take advantage of a stable isolated rat atrial preparation that allows, for the first time, long-term study of synthesis, trafficking, targeting, and secretion of ANF and BNP by adult atrial muscle. Three model stimuli of secretion were used: increased intra-atrial pressure, endothelin-1 (ET-1), and phenylephrine (PE), representing mechanical, hormonal, and alpha1-adrenergic stimuli, respectively. To gain further insight into the secretory process under basal and agonist induced secretion, we employed agents known to inhibit protein synthesis (cycloheximide) or to interfere with the vectorial transport of protein targeted for secretion (brefeldin A and monensin). All these agents induced significant changes in ANF and BNP release. Cycloheximide decreased natriuretic peptide secretion under basal and stimulated conditions. Brefeldin A dramatically increased basal as well as stimulated secretion of ANF and BNP. Monensin partially decreased basal ANF and BNP secretion and completely blocked stimulated secretion. None of these agents modified proteolytic processing as assessed by reverse-phase HPLC analysis. Double-label pulse-chase experiments using [3H]- and [14C]leucine demonstrated that the secretory response to ET-1, in contrast to the response to muscle stretch, is based on peptide other than newly synthesized or relatively newly stored ANF. It is concluded that, in adult atrial cardiocytes, ANF and BNP are sorted to constitutive and regulated pathways in a manner that is substantially unique for atrial cardiocytes. In particular, it appears that basal and stimulated ANF and BNP secretion may have a large "constitutive-like" component, as previously defined in other endocrine systems. This type of secretion is based on the preferential release of hormone through vesicles arising from immature secretory granules. The capacity of the atria to release ANF and BNP in response to stimuli, therefore, may depend more on stimulation of the rate of formation of immature granules than on the amount of stored hormone. PMID- 10362679 TI - Effect of sinoaortic denervation on frequency-domain estimates of baroreflex sensitivity in conscious cats. AB - In animals and humans, baroreceptor modulation of the sinus node in daily life can be studied by identification of the number of sequences in which systolic blood pressure (SBP) and pulse interval (PI) linearly decrease or increase for several beats. It is also studied by power spectral analysis of SBP and PI in regions where their powers are coherent, although, in contrast to the sequence method, whether this frequency-domain method specifically reflects the baroreceptor-heart rate reflex has not been adequately tested. We recorded intra arterial BP for approximately 3.5 h in eight conscious cats, first intact and then 7-10 days after sinoaortic denervation (SAD). Sensitivity of baroreceptor heart rate reflex was assessed in 120-s segments by the square root of the ratio of PI and SBP spectral powers (alpha) in the regions around 0.1 (MF) and 0.3 (HF) Hz, and coherence between PI and SBP spectral powers in MF and HF regions was computed. SAD increased overall SBP variability and reduced PI variability throughout the frequency range examined. SAD markedly reduced (P < 0.01) both alpha-MF (-65.6%) and alpha-HF (-79. 9%) and consistently reduced the number of coherent segments [i.e., where coherence (K2) > 0.5] and average coherence values in the MF region. In the HF region, however, SAD did not alter the number of coherent segments, and although average coherence value throughout the HF band was reduced, in restricted portions of the band (different between animals), a high coherence value survived denervation. No significant changes were seen in any measured variables in five sham-operated cats. Thus the frequency-domain method specifically reflects baroreflex modulation of heart rate in the MF region only. In the HF region, in contrast, baroreflex and nonbaroreflex influences on the sinus node both contribute to a variable degree to determination of heart rate responses to BP oscillations. If used to study baroreflex function in daily life, this method should use the coefficient derived from MF data. PMID- 10362680 TI - Regional ischemia increases sensitivity of left ventricular relaxation to volume in pigs. AB - Regional ischemia impairs early diastolic filling due, in part, to changes in left ventricular relaxation. This study uses open-chest pigs instrumented with high-fidelity pressure transducers to investigate the effect of regional ischemia on the active component of relaxation independent of the passive effects of filling and the effect of left ventricular filling and stretch on the rate, duration, and extent of relaxation. During regional ischemia, active relaxation was impaired in the nonfilling ventricle, with a slower rate of relaxation. Stretching the myocardium as the ventricle fills slows the rate of relaxation more during regional ischemia than during normal perfusion, reflecting an increased sensitivity to stretch due to filling and an increased dependence of relaxation on volume. The duration of relaxation depends on the effect of regional ischemia on the end-diastolic pressure-volume relation. Stronger baseline contractile function results in an upward shift in the end-diastolic pressure-volume relation during regional ischemia and no net effect on the duration of relaxation. If this curve is shifted upward, the duration of relaxation shortens. All these effects combine to reduce the atrioventricular pressure gradient and left ventricular filling during regional ischemia. PMID- 10362681 TI - Cardiovascular phenotype and temperature control in mice lacking thyroid hormone receptor-beta or both alpha1 and beta. AB - We have used a telemetry system to record heart rate, body temperature, electrocardiogram (ECG), and locomotor activity in awake, freely moving mice lacking thyroid hormone receptor (TR)-beta or TR-alpha1 and -beta (TR alpha1/beta). The TR-alpha1/beta-deficient mice had a reduced heart rate compared with wild-type controls. The TR-beta-deficient mice showed an elevated heart rate, which, however, was unresponsive to thyroid hormone treatment regardless of hormonal serum levels. ECG revealed that the TR-beta-deficient mice had a shortened Q-Tend time in contrast to the TR-alpha1/beta-deficient mice, which exhibited prolonged P-Q and Q-Tend times. Mental or pharmacological stimulation of the sympathetic nervous system resulted in a parallel increase in heart rate in all animals. A single injection of a nonselective beta-adrenergic-receptor blocker resulted in a parallel decrease in all mice. The TR-alpha1/beta-deficient mice also had a 0.4 degrees C lower body temperature than controls, whereas no difference was observed in locomotor activity between the different strains of mice. Our present and previous results support the hypothesis that TR-alpha1 has a major role in determining heart rate under baseline conditions and body temperature and that TR-beta mediates a hormone-induced increase in heart rate. PMID- 10362682 TI - Mechanical activity in heart regulates translation of alpha-myosin heavy chain mRNA but not its localization. AB - Mechanical inactivity depresses protein expression in cardiac muscle tissue and results in atrophy. We explore the mechanical transduction mechanism in spontaneously beating neonatal rat cardiomyocytes expressing the alpha-myosin heavy chain (alpha-MyHC) isoform by interfering with cross-bridge function [2,3 butanedione monoxime (BDM), 7.5 mM] without affecting cell calcium. The polysome content and alpha-MyHC mRNA levels in fractions from a sucrose gradient were analyzed. BDM treatment blocked translation at initiation (162 +/- 12% in the nonpolysomal RNA fraction and 43 +/- 6% in the polysomal fraction, relative to control as 100%; P < 0.05). There was an increase in alpha-MyHC mRNA from the nonpolysomal fraction (120.5 +/- 7.7%; P < 0.05 compared with control) with no significant change in the heavy polysomes. In situ hybridization of alpha-MyHC mRNA was used to estimate message abundance as a function of the distance from the nucleus. The mRNA was dispersed through the cytoplasm in spontaneously beating cells as well as in BDM-treated cells (no significant difference). We conclude that direct inhibition of contractile machinery, but not calcium, regulates initiation of alpha-MyHC mRNA translation. However, calcium, not pure mechanical signals, appears to be important for message localization. PMID- 10362684 TI - Ischemic preconditioning and myocardial hypothermia in rabbits with prolonged coronary artery occlusion. AB - This study tests whether combining regional hypothermia and ischemic preconditioning (IP) provides greater myocardial protection during prolonged coronary artery occlusion (CAO) than either intervention alone, and whether increasing the duration of IP from 5 to 7 min extends the window of protection to include a 2-h CAO. Anesthetized rabbits were randomized to four groups (n = 8 rabbits/group): control (C), hypothermia alone (H), IP alone for two 7-min episodes (IP7), and IP plus hypothermia (H + IP7). To compare differences in IP for 5 versus 7 min, additional rabbits (n = 6) received one 5-min episode of ischemia (IP5). All rabbits got 2 h of CAO and 3 h of reperfusion. In comparison with the infarct size in the control group (72 +/- 4% of the risk zone), infarct size was significantly reduced in H (50 +/- 7%), IP7 (49 +/- 5%), and H + IP7 (42 +/- 6%) (all P < 0.05 vs. control group). IP5 failed to confer protection (67 +/- 5% of the risk zone). Therefore, IP can protect against a 2-h CAO if the IP regimen is increased from 5 to 7 min. The combination therapy significantly improved regional myocardial blood flow in the previously ischemic region to a greater extent than either treatment alone. PMID- 10362683 TI - Cationic amino acid transporter gene expression in cultured vascular smooth muscle cells and in rats. AB - Immunostimulants trigger vascular smooth muscle cells (VSMC) to express the inducible isoform of NO synthase (iNOS) and increased arginine transport activity. Although arginine transport in VSMC is considered to be mediated via the y+ system, we show here that rat VSMC in culture express the cat-1 gene transcript as well as an alternatively spliced transcript of the cat-2 gene. An RT-PCR cloning sequence strategy was used to identify a 141-base nucleotide sequence encoding the low-affinity domain of alternatively spliced CAT-2A and a 138-base nucleotide sequence encoding the high-affinity domain of CAT-2B in VSMC activated with lipopolysaccharide (LPS) in combination with interferon-gamma (IFN). With this sequence as a probe, Northern analyses showed that CAT-1 mRNA and CAT-2B mRNA are constitutively present in VSMC, and the expression of both mRNAs was rapidly stimulated by treatment with LPS-IFN, peaked within 4 h, and decayed to basal levels within 6 h after LPS-IFN. CAT-2A mRNA was not detectable in unstimulated or stimulated VSMC. Arginine transporter activity significantly increased 4-10 h after LPS-IFN. iNOS activity was reduced to almost zero in the absence of extracellular arginine uptake via system y+. Induction of arginine transport seems to be a prerequisite to the enhanced synthesis of NO in VSMC. Moreover, this work demonstrates tissue expression of CAT mRNAs with use of a model of LPS injection in rats. RT-PCR shows that the expression of CAT-1 and CAT 2B mRNA in the lung, heart, and kidney is increased by LPS administration to rats, whereas CAT-2A mRNA is abundantly expressed in the liver independent of LPS treatment. These findings suggest that together CAT-1 and CAT-2B play an important role in providing substrate for high-output NO synthesis in vitro as well as in vivo and implicate a coordinated regulation of intracellular iNOS enzyme activity with membrane arginine transport. PMID- 10362685 TI - Systemic and microcirculatory effects of autologous whole blood resuscitation in severe hemorrhagic shock. AB - Systemic and microcirculatory effects of autologous whole blood resuscitation after 4-h hemorrhagic shock with a mean arterial pressure (MAP) level of 40 mmHg were investigated in 63 conscious Syrian golden hamsters. Microcirculation of skeletal skin muscle and subcutaneous connective tissue was visualized in a dorsal skinfold. Shed blood was retransfused within 30 min after 4 h. Animals were grouped into survivors in good (SG) and poor condition (SP) and nonsurvivors (NS) according to 24-h outcome after resuscitation and studied before shock, during shock (60, 120, and 240 min), and 30 min and 24 h after resuscitation. Microvascular and interstitial PO2 values were determined by phosphorescence decay. Shock caused a significant increase of arterial PO2 and decrease of PCO2, pH, and base excess. In the microcirculation, there was a significant decrease in blood flow (QB), functional capillary density (FCD; capillaries with red blood cell flow), and interstitial PO2 [1.8 +/- 0.8 mmHg (SG), 1.3 +/- 1.3 mmHg (SP), and 0.9 +/- 1.1 mmHg (NS) vs. 23.0 +/- 6.1 mmHg at control]. Blood resuscitation caused immediate MAP recompensation in all animals, whereas metabolic acidosis, hyperventilation, and a significant interstitial PO2 decrease (40-60% of control) persisted. In NS (44.4% of the animals), systemic and microcirculatory alterations were significantly more severe both in shock and after resuscitation than in survivors. Whereas in SG (31.8% of the animals) there was only a slight (15-30%) but still significant impairment of microscopic tissue perfusion (QB, FCD) and oxygenation at 24 h, SP (23.8% of the animals) showed severe metabolic acidosis and substantial decreases (>/=50%) of FCD and interstitial PO2. FCD, interstitial PO2, and metabolic state were the main determinants of shock outcome. PMID- 10362686 TI - Increases in oxygen tension stimulate expression of ICAM-1 and VCAM-1 on human endothelial cells. AB - Leukocyte infiltration plays a major role in ischemia-associated organ dysfunction and damage. A crucial step for extravasation of white blood cells is binding of leukocyte beta-integrins to endothelial adhesion molecules intercellular adhesion molecule-1 (ICAM-1) and vascular adhesion molecule-1 (VCAM 1). To test for direct effects of oxygen on this process we studied ICAM-1 and VCAM-1 expression in human dermal microvascular and umbilical vein endothelial cells (EC) exposed to different oxygen tensions in the absence or presence of tumor necrosis factor-alpha (TNF-alpha). Hypoxia (95% N2-5% CO2) resulted in a downregulation of basal but not TNF-alpha-induced expression of ICAM-1 and VCAM 1. Subsequent rises in oxygen (21, 40, or 95% O2) led to marked increase of ICAM 1 and VCAM-1 cell surface and mRNA expression in both EC types, which after 16 h amounted to about one-third to one-half of maximal TNF-alpha-induced expression. This increase was greatest after 0.5-h hypoxia and was blunted with prolonged hypoxic preincubation. Exposure of cells preincubated under "normoxic" (21% O2) conditions to hyperoxia (40 or 95% O2) also enhanced expression of both adhesion molecules, but the increase was lower than in cells preexposed to hypoxia. The nitric oxide synthesis inhibitor NG-nitro-L-arginine methyl ester (L-NAME) enhanced ICAM-1 and VCAM-1 expression under basal and hypoxic conditions, but in the presence of L-NAME, levels in reoxygenated cells were not higher than basal levels. Moreover, the oxygen-induced rise could be mimicked by addition of H2O2 to normoxic cells, and the oxygen-induced expression of VCAM-1 but not of ICAM-1 was inhibited by addition of the free radical scavengers superoxide dismutase, N acetyl-L-cysteine, and pyrrolidinedithiocarbamate. These data indicate that an increase in oxygen availability stimulates ICAM-1 and VCAM-1 expression on micro- and macrovascular EC, which may contribute to adhesion and transmigration of different leukocyte populations in ischemia-reperfusion injuries. PMID- 10362687 TI - Brain O2 consumption and glutamate release during hypoglycemic coma in piglets are temperature sensitive. AB - Hypoglycemic injury in the mature brain is mediated by excitotoxicity, which is worsened by disordered cellular energy metabolism. The role of excitotoxicity in relation to brain energy metabolism during hypoglycemia has not been studied in the immature brain. Brain oxygen consumption (CMRO2) increases during hypoglycemia in piglets, whereas CMRO2 decreases in adult pig models. We tested the hypothesis that increased CMRO2 during hypoglycemic coma is temperature dependent and coincides with increased excitatory amino acids (EAA). We measured cerebral blood flow (CBF), CMRO2, and cortical microdiaysate EAA in pentobarbital anesthetized piglets during hypoglycemic coma and during 2 h of recovery and in normoglycemic controls. In warmed animals brain temperature was kept normothermic (38.5 degrees C). In unwarmed animals brain temperature was allowed to fall (37.6 degrees C). During hypoglycemia CBF increased similarly in warmed animals and unwarmed animals; CMRO2 increased in warmed animals but not unwarmed animals. Glutamate increased during coma and increased more in warmed animals than unwarmed animals but normalized quickly during recovery. EEG recovered earlier in unwarmed animals. We conclude that during a hypoglycemic coma in the immature brain, CMRO2 and glutamate are increased in a temperature-dependent manner. PMID- 10362688 TI - Effects of calcitonin gene-related peptide on vascular resistance in rats: role of sex steroids. AB - It has been demonstrated in reflex-intact animals that the sensitivity to calcitonin gene-related peptide (CGRP) is increased during pregnancy and that this action is mediated by sex steroids but not by nitric oxide (NO). We assessed the effects of CGRP in the following groups of anesthetized ganglion-blocked rats: 1) pregnant, 2) ovariectomized, and 3) ovariectomized and treated with estradiol and progesterone. Changes in mean arterial pressure (MAP) were assessed after the administration of varying doses of CGRP. Decreases in MAP after CGRP administration were significantly greater in pregnant rats and ovariectomized rats administered sex steroids than in ovariectomized controls. The CGRP antagonist CGRP8-37 produced a pressor response of similar magnitude in both pregnant and ovariectomized rats. We also assessed the effects of CGRP and the modulating role of NO in the isolated uterine vascular bed preparation. CGRP reduced perfusion pressure to a greater degree in ovariectomized animals treated with sex steroids than in ovariectomized animals. This response was attenuated by pretreatment with an NO synthesis inhibitor. CGRP8-37 produced a similar increase in perfusion pressure in both groups. We conclude that 1) the increased vascular sensitivity observed during pregnancy or after treatment with sex steroids is in part mediated by NO, and 2) CGRP8-37 has a vasoconstrictor action of its own. PMID- 10362689 TI - NO modulates myocardial O2 consumption in the nonhuman primate: an additional mechanism of action of amlodipine. AB - Recent evidence from our laboratory and others suggests that nitric oxide (NO) is a modulator of in vivo and in vitro oxygen consumption in the murine and canine heart. Therefore, the goal of our study was twofold: to determine whether NO modulates myocardial oxygen consumption in the nonhuman primate heart in vitro and to evaluate whether the seemingly cardioprotective actions of amlodipine may involve an NO-mediated mechanism. Using a Clark-type O2 electrode, we measured oxygen consumption in cynomologous monkey heart at baseline and after increasing doses of S-nitroso-N-acetylpenicillamine (SNAP; 10(-7)-10(-4) M), bradykinin (10( 7)-10(-4) M), ramiprilat (10(-7)-10(-4) M), and amlodipine (10(-7)-10(-5) M). SNAP (-38 +/- 5.8%), bradykinin (-19 +/- 3.9%), ramiprilat (-28 +/- 2.3%), and amlodipine (-23 +/- 4.5%) each caused significant (P < 0.05) reductions in myocardial oxygen consumption at their highest dose. Preincubation of tissue with nitro-L-arginine methyl ester (10(-4) M) blunted the effects of bradykinin (-5.4 +/- 3.2%), ramiprilat (-4.8 +/- 5.0%), and amlodipine (-5.3 +/- 5.0%) but had no effect on the tissue response to SNAP (-38 +/- 5.8%). Our results indicate that NO can reduce oxygen consumption in the primate myocardium in vitro, and they support a role for the calcium-channel blocker amlodipine as a modulator of myocardial oxygen consumption via a kinin-NO mediated mechanism. PMID- 10362690 TI - Species-dependent hemodynamic effects of adenosine A3-receptor agonists IB-MECA and Cl-IB-MECA. AB - The purpose of this study was to compare the hemodynamic effects of the adenosine A3-receptor agonists N6-(3-iodobenzyl)-9-[5-(methylcarbamoyl)-beta-D ribofuranosyl]aden ine (IB-MECA) and 2-chloro-N6-(3-iodobenzyl)-9-[5 (methylcarbamoyl)-beta-D-ribofu ranosy l]adenine (Cl-IB-MECA) in isolated rat and rabbit hearts and in the intact, open-chest pig. Isolated hearts perfused with Krebs-Henseleit buffer at a constant pressure (70 mmHg) were treated with 50 nM of either IB-MECA or Cl-IB-MECA. Neither IB-MECA nor Cl-IB-MECA altered ventricular function or heart rate in the isolated rat and rabbit hearts, and neither agent altered coronary flow in the rabbit. However, 2 min of IB-MECA treatment in the isolated rat heart increased coronary flow by 25%, an effect that did not exhibit tachyphylaxis. The IB-MECA-induced coronary dilation was only partially attenuated by the adenosine A3-receptor antagonist MRS-1191 (50 nM). IB-MECA-induced coronary dilation was completely blocked by the adenosine A2a-receptor antagonist 7-(2-phenylethyl)-5-amino-2-(2-furyl)-pyrazolo-[4,3-e] 1,2, 4-triazolo[1,5-c]pyrimidine (Sch-58261, 50 nM). Cl-IB-MECA (50 nM) did not increase coronary flow in the rat, but 100 nM did increase flow by 18%. In pentobarbital sodium-anesthetized pigs IB-MECA (5 micrograms/kg iv) decreased systemic blood pressure and increased pulmonary artery pressure, effects that did exhibit tachyphylaxis. These results illustrate that adenosine A3-receptor agonists produce species-dependent effects, which in the rat heart appear to be caused by adenosine A2a-receptor activation. PMID- 10362691 TI - Cannabinoid CB1 receptor of cat cerebral arterial muscle functions to inhibit L type Ca2+ channel current. AB - The CB1 subtype of the cannabinoid receptor is present on neurons in the brain and mediates the perceptual effects of Delta9-tetrahydrocannabinol and other cannabinoids. We found that cat cerebral arterial smooth muscle cells (VSMC) contain the protein for the CB1 receptor and express a cDNA that has >98% amino acid homology to the CB1 cDNA expressed in rat and human neurons. Activation of the CB1 cannabinoid receptor has been shown to decrease the opening of N-type voltage-gated Ca2+ channels in neurons through a pertussis toxin-sensitive GTP binding protein. In the present study we tested the hypothesis that activation of the cannabinoid CB1 receptor in cerebral VSMC inhibits voltage-gated Ca2+ channels and results in cerebral vasodilation. The predominant Ca2+ current identified in cat cerebral VSMC is a voltage-gated, dihydropyridine-sensitive, L type Ca2+ current. The cannabimimetic drug WIN-55,212-2 (10-100 nM) induced concentration-dependent inhibition of peak L-type Ca2+ current, which reached a maximum of 82 +/- 4% at 100 nM (n = 14). This effect was mimicked by the putative endogenous CB1-receptor agonist anandamide, which produced a concentration related reduction of peak L-type Ca2+ current with a maximum inhibition (at 300 nM) of 39 +/- 4% (n = 12). The inhibitory effects of both ligands on peak L-type Ca2+ currents were abolished by pertussis toxin pretreatment and application of the CB1-receptor antagonist SR-141716A (100 nM, n = 5). Both WIN-55,212-2 and anandamide produced concentration-dependent relaxation of preconstricted cerebral arterial segments that was abolished by SR-141716A. These results indicate that the CB1 receptor is expressed in cat cerebral VSMC and that the cerebral vasculature is one of the targets for endogenous cannabinoids. These findings suggest that the CB1 receptor and its endogenous ligand may play a fundamental role in the regulation of cerebral arterial tone and reactivity by modulating the influx of Ca2+ through L-type Ca2+ channels. PMID- 10362692 TI - Lipoxygenase metabolism of arachidonic acid in ischemic preconditioning and PKC induced protection in heart. AB - We tested the hypothesis that activation of the 12-lipoxygenase (12-LO) pathway of arachidonic acid metabolism contributes to the protective effect of protein kinase C (PKC) activation and ischemic preconditioning (PC), and we report, in perfused rat heart, that both PC and the PKC activator 1,2-dioctanoyl-sn-glycerol (DOG) confer a similar protective effect and stimulate a comparable accumulation of 12-LO metabolites. The 12-LO product, 12(S)-hydroxyeicosatetraenoic acid [12(S)-HETE], was increased in DOG-treated (22.8 +/- 4.4 ng/g wet wt) and PC hearts (26.8 +/- 5.5 ng/g wet wt) compared with control (13.8 +/- 2.1 ng/g wet wt, P < 0. 05), and this increase was blocked by 12-LO or PKC inhibitors. Both DOG pretreatment and PC improved recovery of left ventricular developed pressure (LVDP) nearly twofold after 20 min of ischemia; this improvement was blocked by 12-LO inhibitors and was mimicked by infusion of 12-hydroperoxyeicosatetraenoic acid [12(S)-HpETE; 67 +/- 6% recovery of LVDP vs. 35 +/- 3% for untreated hearts]. Also, the protection afforded by 12(S)-HpETE, as well as by PC, was attenuated by the K+-channel blocker 5-hydroxydecanoate, suggesting that the downstream mechanisms of 12(S)-HpETE-mediated protection are similar to PC. Furthermore, PC stimulates 12-LO metabolism in perfused rabbit heart, and 12-LO inhibition blocks PC-induced cardioprotection. Thus the data suggest that 12-LO metabolism plays an important role in cardioprotection. PMID- 10362693 TI - End-systolic myocardial stiffness is a load-independent index of contractility in stage 24 chick embryonic heart. AB - Cardiac morphogenesis and function are interrelated during cardiovascular development. We evaluated the effects of acute alteration of loading condition to chick embryonic ventricular contractility using end-systolic myocardial stiffness based on the incremental elastic modulus concept. End-systolic stress-strain relations including geometric factor and end-systolic myocardial stiffness were determined from the simultaneous measurement of ventricular pressure and chamber dimension in the following four groups of stage 24 White Leghorn chick embryos: volume infusion (n = 9), conotruncal occlusion (n = 9), calcium suffusion (n = 10), and verapamil suffusion (n = 8). The end-systolic stress-strain relationship was linear in each embryo. There was no correlation between end-systolic myocardial stiffness and end-systolic stress. End-systolic myocardial stiffness increased with calcium suffusion (P < 0.05 vs. volume infusion). The geometric factor increased after verapamil suffusion (P < 0.05). End-systolic myocardial stiffness normalized by geometric factor was not changed by alteration of preload or afterload, increased after calcium suffusion, and decreased after verapamil administration (P < 0.05). These results suggest that normalized end-systolic myocardial stiffness is a load-independent index of ventricular contractility in the developing embryonic chick ventricle. PMID- 10362694 TI - Participation of PI3K and atypical PKC in Na+-K+-pump stimulation by IGF-I in VSMC. AB - The activity of the Na+-K+-pump is intricately linked to the maintenance of vascular tone. Here we demonstrate that insulin-like growth factor I (IGF-I) increases Na+-K+-pump activity in the vascular smooth muscle cell (VSMC) clone A7r5 in a time- and dose-dependent manner. This stimulatory effect of IGF-I was prevented by the tyrosine kinase inhibitor genistein (5 microM) and by the specific phosphatidylinositol 3-kinase (PI3K) inhibitors wortmannin (100 nM) and LY-294002 (25 microM). IGF-I activated a wortmannin-sensitive PI3K and its purported effector, the atypical protein kinase C (PKC)-zeta. Stimulation of PKC zeta was prevented by the generic PKC inhibitor GF109203x (bisindolylmaleimide, 10 microM). Downregulation of diacylglycerol-sensitive (conventional and novel) PKCs by 24-h pretreatment with 1 microM phorbol 12-myristate 13-acetate had no effect on IGF-I-stimulated Na+-K+-pump activity. Similarly, inhibition of only conventional and novel PKCs with GF109203x (1 microM) had no effect on IGF-I stimulated Na+-K+-pump activity. In contrast, a concentration of GF109203x (10 microM) that also inhibits the atypical PKCs abolished Na+-K+-pump stimulation by IGF-I. Neither the Na+-K+-2Cl- cotransporter inhibitor bumetanide (100 microM) nor the Na+/H+ exchanger inhibitor HOE-694 (5 microM) affected the Na+-K+-pump stimulation by IGF-I, suggesting that a rise in intracellular Na+ concentration is not necessary for increased Na+-K+-pump activity. These results suggest that IGF-I directly stimulates the Na+-K+ pump via a signaling pathway involving PI3K and atypical PKC (zeta). PMID- 10362695 TI - Rapid tachyphylaxis to hemodynamic effects of PACAP-27 after inhibition of nitric oxide synthesis. AB - The vasodilator effects of pituitary adenylate cyclase-activating polypeptide (PACAP)-27 are subject to tachyphylaxis in rats treated with the nitric oxide (NO) synthase inhibitor NG-nitro-L-arginine methyl ester (L-NAME). We examined whether this tachyphylaxis could be prevented by administration of the putative endothelium-derived nitrosyl factor S-nitroso-L-cysteine (L-SNC) and whether L SNC may exert its effects via increases in cGMP levels in vascular smooth muscle. Five doses of PACAP-27 (2 nmol/kg iv) produced pronounced vasodilator responses in saline-treated rats. These responses were not subject to tachyphylaxis. The first injection of PACAP-27 (2 nmol/kg iv) in L-NAME-treated (50 micromol/kg iv) rats produced vasodilator responses similar to those in saline-treated rats, whereas subsequent injections produced progressively smaller responses. The injection of L-SNC (1,200 nmol/kg iv) before each injection of PACAP-27 prevented tachyphylaxis to the Gs protein-coupled receptor agonist in L-NAME-treated rats, whereas equihypotensive doses of the NO donor sodium nitroprusside (100 micrograms/kg iv) did not. The injection of the membrane-permeant cGMP analog 8 (4-chlorophenylthio)guanosine 3',5'-cyclic monophosphate (8-CPT-cGMP; 30 micromol/kg iv) to L-NAME-treated rats restored resting hemodynamic values to pre L-NAME levels but did not prevent the development of tachyphylaxis to PACAP-27. These results suggest that nitrosyl factors prevent the development of tachyphylaxis to the hemodynamic actions of PACAP-27. These nitrosyl factors may act independently of their ability to generate cGMP in vascular smooth muscle. PMID- 10362696 TI - Naloxone reverses inhibitory effect of electroacupuncture on sympathetic cardiovascular reflex responses. AB - Acupuncture and electroacupuncture (EA) have been used in traditional Chinese medicine to treat a wide range of diseases and conditions, including angina pectoris and myocardial infarction. In a feline model of reflex-induced reversible myocardial ischemia, electrical stimulation of the median nerves to mimic EA (Neiguan acupoint) significantly improved ischemic dysfunction, secondary to an inhibitory effect of EA on reflex pressor effects evoked by bradykinin (BK). The central mechanism of EA's inhibitory effect in this model is unknown. Accordingly, in alpha-chloralose-anesthetized cats, BK (10 micrograms/ml) was applied to the gallbladder to elicit a cardiovascular reflex response that significantly (P < 0.05) increased arterial blood pressure and heart rate; normalized systolic wall thickening (%WTh) of the left ventricle, measured by ultrasonic single-crystal sonomicrometer, increased by 31 +/- 11% (P < 0.05). After ligation of a side branch of the left anterior descending coronary artery, the reflex pressor response to BK resulted in a significant decrease of %WTh (-32 +/- 6%) in the ischemic region. When bilateral EA of the Neiguan acupoints was performed, the pressor response to BK was inhibited and regional myocardial function was significantly improved (+19 +/- 20%). The inhibitory effects of EA on blood pressure and %WTh were reversed by intravenous injection of naloxone (0.4 mg/kg; n = 9) or microinjection of naloxone (10 nM in 0.1 microliter/site; n = 14) into the rostral ventrolateral medulla (rVLM). Thus %WTh with intravenous naloxone was reduced to -13 +/- 29% (P<0.05) during stimulation of the gallbladder. Our results indicate that the inhibitory effect of EA on the BK-induced pressor response and the consequent improvement of ischemic dysfunction is dependent on the activation of opioid receptors, specifically receptors located in the rVLM. PMID- 10362697 TI - Enhanced DNA fragmentation in the thymus of spontaneously hypertensive rats. AB - The mechanisms contributing to organ injury in hypertension have been incompletely defined. The thymus gland of the spontaneously hypertensive rat (SHR) shows significant atrophy at the age of 15 wk compared with its normotensive control, the Wistar-Kyoto rat (WKY). The aim of the present study was to examine the thymus of SHR for evidence of DNA nicking as one of the mechanisms for thymic atrophy. SHR and WKY were subjected to adrenalectomy or sham surgery at 12 wk and studied at 15 wk. Adrenalectomy served to normalize the blood pressure in the SHR. DNA nicking was detected by in situ nick-end labeling (ISEL) of fixed tissue sections. Tissue sections were treated with proteolysis, and terminal deoxyribonucleotidyl transferase was used to incorporate biotinylated deoxynucleotides into DNA nick end in situ. Separately, DNA fragmentation was evaluated by measuring the level of released mono- and oligonucleosomes to the cytoplasm. A higher number of thymic ISEL-positive cells and a higher level of cytoplasmic mono- and oligonucleosomes were observed in SHR than in WKY. After adrenalectomy the enhanced level of ISEL and cytoplasmic mono- and oligonucleosomes in SHR was reduced to the level in WKY. Dexamethasone treatment (0.05 mg. kg-1. day-1) in WKY serves to decrease the thymus weight and significantly elevate the level of mono- and oligonucleosomes. Thus increased DNA fragmentation represents one of the mechanisms associated with thymic atrophy, a feature that reflects immune suppression in SHR. PMID- 10362698 TI - Differential regulation of cardiac expression of IL-6 and TNF-alpha by A2- and A3 adenosine receptors. AB - The proinflammatory cytokines tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 have been implicated in the development of congestive heart failure. Adenosine inhibits the expression of TNF-alpha and IL-6 in macrophages. We determined the effect of adenosine on cytokine expression in rat cardiomyocytes and trabecular muscles obtained from patients with cardiomyopathy. In myocytes, adenosine suppressed TNF-alpha mRNA by 40% (P < 0.05) and induced a 4.7-fold increase in IL-6 mRNA (P < 0.05) with a twofold increase in IL-6 protein release (P < 0.001). The effect on TNF-alpha could be replicated by A2 agonist. The effect on IL-6 could be replicated by A3 agonist, but not by A1 and A2 agonists, and was completely suppressed by A3 antagonist. In human trabecular muscles, A2 agonist suppressed TNF-alpha mRNA by 60% (P < 0.05), but adenosine had no effect on IL-6. In the failing heart, IL-6 was immunolocalized to inflammatory cells. Thus A2 and A3 receptors differentially regulate cardiac expression of TNF-alpha and IL-6. Rat cardiomyocytes and the failing human heart respond differently to adenosine. PMID- 10362699 TI - Cardiac myosin heavy chains lacking the light chain binding domain cause hypertrophic cardiomyopathy in mice. AB - Myosin is a chemomechanical motor that converts chemical energy into the mechanical work of muscle contraction. More than 40 missense mutations in the cardiac myosin heavy chain (MHC) gene and several mutations in the two myosin light chains cause a dominantly inherited heart disease called familial hypertrophic cardiomyopathy. Very little is known about the biochemical defects in these alleles and how the mutations lead to disease. Because removal of the light chain binding domain in the lever arm of MHC should alter myosin's force transmission but not its catalytic function, we tested the hypothesis that such a mutant MHC would act as a dominant mutation in cardiac muscle. Hearts from transgenic mice expressing this mutant myosin are asymmetrically hypertrophied, with increases in mass primarily restricted to the cardiac anterior wall. Histological examination demonstrates marked cellular hypertrophy, myocyte disorganization, small vessel coronary disease, and severe valvular pathology that included thickening and plaque formation. Skinned myocytes and multicellular preparations from transgenic hearts exhibited decreased Ca2+ sensitivity of tension and decreased relaxation rates after flash photolysis of diazo 2. These experiments demonstrate that alterations in myosin force transmission are sufficient to trigger the development of hypertrophic cardiomyopathy. PMID- 10362700 TI - PDGF-A expression correlates with blood pressure and remodeling in 1K1C hypertensive rat arteries. AB - We previously demonstrated remodeling of large and small arteries in angiotensin II-treated rats, paralleled by an increased expression of platelet-derived growth factor (PDGF)-A chain mRNA in large arteries. Both remodeling and PDGF-A expression were associated with elevation of blood pressure rather than a direct effect of angiotensin II. To further delineate the role of PDGF-A and elevated blood pressure, we assessed the level of PDGF-A and -B mRNA and protein in the wall of large as well as small arteries in the one-kidney, one-clip (1K1C) hypertensive rat, a non-renin-dependent model of hypertension. Fourteen days after renal artery stenosis, the thoracic aorta and both femoral arteries were collected from 1K1C rats (n = 8) and uninephrectomized controls (n = 8) and immediately processed for morphological measurement, immunohistochemistry, RT PCR, and Western blotting. Systolic blood pressure was significantly elevated in hypertensive rats (202 +/- 26 mmHg) compared with control rats (122 +/- 7.9 mmHg) and was accompanied by arterial hypertrophy in both aorta and femoral arteries. The mRNA for PDGF-A chain was increased threefold in the thoracic aorta (P < 0.05) of 1K1C rats, whereas the message for PDGF-B was not significantly changed in hypertensive versus control animals. A higher staining of the intima-media was observed by using an anti-PDGF-A chain polyclonal antibody on paraffin-embedded sections. Western blot results indicated an approximately 2-fold increase in PDGF A protein in aortic and femoral wall of the 1K1C rats. The results showed that both the mRNA and protein for PDGF-A chain are increased and well correlated with the blood pressure and wall area, suggesting a direct effect of elevated pressure on PDGF synthesis, which, in turn, may affect the onset and progression of vascular hypertrophy. PMID- 10362701 TI - CaM kinase augments cardiac L-type Ca2+ current: a cellular mechanism for long Q T arrhythmias. AB - Early afterdepolarizations (EAD) caused by L-type Ca2+ current (ICa, L) are thought to initiate long Q-T arrhythmias, but the role of intracellular Ca2+ in these arrhythmias is controversial. Rabbit ventricular myocytes were stimulated with a prolonged EAD-containing action potential-clamp waveform to investigate the role of Ca2+/calmodulin-dependent protein kinase II (CaM kinase) in ICa,L during repolarization. ICa,L was initially augmented, and augmentation was dependent on Ca2+ from the sarcoplasmic reticulum because the augmentation was prevented by ryanodine or thapsigargin. ICa,L augmentation was also dependent on CaM kinase, because it was prevented by dialysis with the inhibitor peptide AC3-I and reconstituted by exogenous constitutively active CaM kinase when Ba2+ was substituted for bath Ca2+. Ultrastructural studies confirmed that endogenous CaM kinase, L-type Ca2+ channels, and ryanodine receptors colocalized near T tubules. EAD induction was significantly reduced in current-clamped cells dialyzed with AC3-I (4/15) compared with cells dialyzed with an inactive control peptide (11/15, P = 0.013). These findings support the hypothesis that EADs are facilitated by CaM kinase. PMID- 10362703 TI - Greater erythrocyte deformability in world-class endurance athletes. AB - Because athletes during endurance events require rapid uptake of oxygen, the ability of red blood cells (RBC) to move through capillaries may limit performance. Using ektacytometry, we determined whether RBC deformability (RCD) differed between elite road cyclists (n = 9) and sedentary controls (n = 5). Density profiles and standard hematological measurements were also performed. The deformability index (DI) was higher in the cyclists (0.723 +/- 0.027) compared with that in controls (0.619 +/- 0.040, P < 0.001). Cyclists also had a larger percentage of low-density RBCs (P < 0. 001), and mean cell volume (MCV) was also higher (P = 0.013). These findings are indicative of a larger proportion of "young" RBCs in the blood of elite cyclists and provide further evidence that the turnover of RBCs in endurance athletes is higher than in the general population. With a younger more deformable RBC population and providing the destruction does not exceed replacement, performance potential should be enhanced. Furthermore, examination of factors that contribute to increased RBC turnover in athletes may help us understand the mechanisms that cause RBC aging. PMID- 10362702 TI - Microvascular endothelial cells remodel cultured adult cardiomyocytes and increase their survival. AB - We investigated the paracrine effect of cardiac microvascular endothelial cells (MVEC) on cultured adult rat cardiomyocytes (ARC). ARC were exposed for 8 days to serum-free medium (CM) conditioned by MVEC. Controls were grown in FCS or FCS free medium. Protein synthesis of CM-stimulated ARC increased twofold versus 5% FCS-stimulated cells until day 8. Seventy-nine percent of CM-treated myocytes survived, whereas only twenty-four percent of FCS-free ARC retained viability. The phenotype of myocytes exposed to CM was different from control. Analysis by confocal laser microscopy of CM-stimulated myocytes showed actin staining throughout the whole cell body up to the peripheral extensions, with concomitant appearance of myomesin in a cross-striated pattern. The reexpression of fetal alpha-smooth muscle actin determined immunohistochemically and by Western blot increased from day 6 in CM-treated cells, whereas ARC grown in up to 20% serum were negative. These effects could not be mimicked by any of the other cardioactive substances tested here, indicating a novel trophic factor in CM. PMID- 10362704 TI - Plasma volume expansion with solutions of hemoglobin, albumin, and Ringer lactate in sheep. AB - We have measured plasma volume expansion (Evans blue and hematocrit changes) and hemodynamic responses in conscious hemorrhaged and normovolemic splenectomized sheep after a 30-min infusion of either 20 ml/kg of diaspirin cross-linked hemoglobin (DCLHb), 20 ml/kg of human albumin (Alb), or 60 ml/kg of a solution of Ringer lactate (RL). All regimens expanded blood volume and increased blood pressure and cardiac output after hemorrhage. However, only 15 +/- 3% of the infused volume of RL was evident as intravascular expansion 10-min postinfusion, compared with 67 +/- 16% and 139 +/- 139% for Alb and DCLHb, respectively. DCLHb infusions were associated with higher blood pressures and lower cardiac outputs compared with RL and Alb infusions, but the increased oxygen content of blood with DCLHb resulted in systemic delivery of oxygen similar to that of the other infusions. These differences in hemodynamics and vascular volume continued for 6 h, and at 24 h vascular volume and all hemodynamics were similar in all three groups. The better volume expansion with DCLHb may be due to greater mobilization of endogenous interstitial protein or reduced transcapillary loss as total intravascular endogenous plasma protein increased after infusion of DCLHb, whereas there was an apparent loss of endogenous intravascular protein after infusions of Alb and RL. Vasoconstriction by DCLHb is one mechanism that could lower blood-to-tissue transport of fluid and protein. In addition to its oxygen carrying capacity and vasoactivity, DCLHb is associated with volume expansion properties out of proportion to its colloid osmotic pressure. PMID- 10362705 TI - Hemodynamic model for analysis of Doppler ultrasound indexes of umbilical blood flow. AB - A hemodynamic model for pulsatile fluid flow in a pressurized thin-walled elastic tube was applied for the computation of volumetric blood flow and velocity profiles for a given set of system parameters at any selected location along the umbilical artery. The velocity profiles over one heart cycle provide the fetal blood flow velocity waveforms (FVW) from which the usual Doppler indexes (DI) can be derived. The model was used for a comprehensive investigation of the correlation between DI and system parameters that reflect the anatomy and physiology of umbilical blood flow. The simulations showed that the radial location of the Doppler measurement is insignificant for the calculated DI, whereas the axial site is important. The analysis showed that decreasing the diameter or increasing the length of the umbilical artery reduces fetal mean blood flow rate and increases the DI. Increasing blood viscosity tends to induce similar patterns, whereas decreasing arterial compliance or increasing blood density decreases the DI with little effect on blood flow rate. Fetal heart rate has a minor effect on both DI and fetal blood flow rate. This study provides insight into the dependence of DI on the anatomic and physiological characteristics of umbilical blood flow. PMID- 10362706 TI - Effect of amiloride analogs on DOCA-salt-induced hypertension in rats. AB - Intracerebroventricular infusions of an amiloride analog, benzamil, reduce blood pressure in several rat models of hypertension. This effect has been attributed to an inhibition of amiloride-sensitive Na+ channels in the brain. This study examines whether intracerebroventricular benzamil would prevent the onset of deoxycorticosterone acetate (DOCA)-salt-induced hypertension in rats and whether this effect correlates with an inhibition of ion transport through the known amiloride-sensitive cation channels at the blood-brain barrier. We also examine whether the effects of benzamil on blood pressure are mediated by a Na+ channel by comparing the effects of different amiloride analogs. Benzamil (0.15 and 0.5 microgram/h icv) did significantly attenuate the increase in blood pressure induced by DOCA treatment. This antihypertensive effect, however, was not associated with an alteration in a blood-brain barrier ion transport as assessed by measurements of blood-to-brain 22Na transport and cerebral spinal fluid Na+ and K+ concentrations. Indeed, intracerebroventricular infusion of dimethyl amiloride, an amiloride analog with low affinity for Na+ channels, also attenuated the increase in blood pressure induced by DOCA-salt treatment. Comparisons of the effects of benzamil, dimethyl amiloride, and 3,4 dichlorobenzamil, another amiloride analog, suggest that these antihypertensive effects are mediated by an inhibition of Na+/Ca2+ exchange in the brain. PMID- 10362707 TI - Parasympathetic modulation of sinoatrial node pacemaker activity in rabbit heart: a unifying model. AB - We have extended our compartmental model [Am. J. Physiol. 266 (Cell Physiol. 35): C832-C852, 1994] of the single rabbit sinoatrial node (SAN) cell so that it can simulate cellular responses to bath applications of ACh and isoprenaline as well as the effects of neuronally released ACh. The model employs three different types of muscarinic receptors to explain the variety of responses observed in mammalian cardiac pacemaking cells subjected to vagal stimulation. The response of greatest interest is the ACh-sensitive change in cycle length that is not accompanied by a change in action potential duration or repolarization or hyperpolarization of the maximum diastolic potential. In this case, an ACh sensitive K+ current is not involved. Membrane hyperpolarization occurs in response to much higher levels of vagal stimulation, and this response is also mimicked by the model. Here, an ACh-sensitive K+ current is involved. The well known phase-resetting response of the SAN cell to single and periodically applied vagal bursts of impulses is also simulated in the presence and absence of the beta-agonist isoprenaline. Finally, the responses of the SAN cell to longer continuous trains of periodic vagal stimulation are simulated, and this can result in the complete cessation of pacemaking. Therefore, this model is 1) applicable over the full range of intensity and pattern of vagal input and 2) can offer biophysically based explanations for many of the phenomena associated with the autonomic control of cardiac pacemaking. PMID- 10362708 TI - Modulation of force-frequency relation by phospholamban in genetically engineered mice. AB - Phospholamban levels regulate cardiac sarcoplasmic reticulum Ca2+ pump activity and myocardial contractility. To determine whether and to what extent phospholamban modulates the force-frequency relation and ventricular relaxation in vivo, we studied transgenic mice overexpressing phospholamban (PLBOE), gene targeted mice without phospholamban (PLBKO), and isogenic wild-type controls. Contractility was assessed by the peak rate of left ventricular (LV) isovolumic contraction (+dP/dtmax), and diastolic function was assessed by both the peak rate (-dP/dtmax) and the time constant (tau) of isovolumic LV relaxation, using a high-fidelity LV catheter. Incremental atrial pacing was used to generate heart rate vs. -dP/dtmax (force-frequency) relations. Biphasic force-frequency relations were produced in all animals, and the critical heart rate (HRcrit) was taken as the heart rate at which dP/dtmax was maximal. The average LV +dP/dtmax increased in both PLBKO and PLBOE compared with their isogenic controls (both P < 0.05). The HRcrit for LV +dP/dtmax was significantly higher in PLBKO (427 +/- 20 beats/min) compared with controls (360 +/- 18 beats/min), whereas the HRcrit in PLBOE (340 +/- 30 beats/min) was significantly lower compared with that in isogenic controls (440 +/- 25 beats/min). The intrinsic heart rates were significantly lower, and the HRcrit and the +/-dP/dtmax at HRcrit were significantly greater in FVB/N than in SvJ control mice. We conclude that 1) the level of phospholamban is a critical negative determinant of the force-frequency relation and myocardial contractility in vivo, and 2) contractile parameters may differ significantly between strains of normal mice. PMID- 10362709 TI - New analytic framework for understanding sympathetic baroreflex control of arterial pressure. AB - The sympathetic baroreflex is an important feedback system in stabilization of arterial pressure. This system can be decomposed into the controlling element (mechanoneural arc) and the controlled element (neuromechanical arc). We hypothesized that the intersection of the two operational curves representing their respective functions on an equilibrium diagram should define the operating point of the arterial baroreflex. Both carotid sinuses were isolated in 16 halothane-anesthetized rats. The vagi and aortic depressor nerves were cut bilaterally. Carotid sinus pressure (CSP) was sequentially altered in 10-mmHg increments from 80 to 160 mmHg while sympathetic efferent nerve activity (SNA) and systemic arterial pressure (SAP) were recorded simultaneously under various hemorrhagic conditions. The mechanoneural arc was characterized by the response of SNA to CSP and the neuromechanical arc by the response of SAP to SNA. We parametrically analyzed the relationship between input and output for each arc using a four-parameter logistic equation model. In baseline states, the two arcs intersected each other at the point at which the instantaneous gain of each arc attained its maximum. Severe hemorrhage lowered the gain and offset of the neuromechanical arc and moved the operating point, whereas the mechanoneural arc remained unchanged. The operating points measured under the closed-loop conditions were indistinguishable from those estimated from the intersections of the two arc curves on the equilibrium diagram. The average root mean square errors of estimate for arterial pressure and SNA were 2 and 3%, respectively. Such an analytic approach could explain a mechanism for the determination of the operating point of the sympathetic baroreflex system and thus helps us integratively understand its function. PMID- 10362710 TI - Branching patterns of intramural coronary vessels determined by microangiography using synchrotron radiation. AB - The intramural coronary artery (IMCA) with a diameter of 50-500 micrometers is critical for blood supply to the inner layers of heart muscle. We introduced digital measurement to microangiography using monochromatic synchrotron radiation and quantified branching patterns of the IMCA, the epicardial coronary artery (EPCA), and the distal ileal artery (DIA). The pre- and postbranching diameters were measured (95-1,275 micrometers) in seven dogs. A typical arterial segment divided into two nearly equivalent branches, and a regression line of daughter-to mother diameter plots was almost identical among the EPCA (y = 0.838x - 16.7 in micrometers), IMCA (y = 0.737x - 2.18), and DIA (y = 0.755x + 8.63). However, a considerable difference was present at a segment where a proximal IMCA branched off from an EPCA (y = 0.182x + 90.2). Moreover, a proximal IMCA diameter had no relationship to the branching order from an EPCA. The precision of this method was confirmed by the good correlation of diameter measurements between two independent observers (r = 0.999, y = 1.02x - 1.07). In conclusion, using digital microangiography we demonstrated that the self-similar branching pattern of coronary arteries was discrete at the connection between the IMCA and EPCA. PMID- 10362711 TI - Modulation of the adaptive response to myocardial ischemia by coexisting disease. PMID- 10362712 TI - Effect of FK506 on ATP-induced intracellular calcium oscillations in cow tracheal epithelium. AB - To elucidate the effect of FK506 on Ca2+ oscillations in airway epithelium, we investigated cultured cow tracheal epithelial cells with a Ca2+ image-analysis system. ATP (1 microM) induced long-lasting Ca2+ oscillations, having nearly constant peak values (300-400 nM) and intervals (20-40 s) in subconfluent cells but not in confluent cells. These responses were gradually attenuated and abolished by the addition of FK506. Rapamycin, which binds the FK506-binding protein (FKBP), likewise inhibited Ca2+ oscillations, whereas cyclosporin A, a calcineurin inhibitor, did not. Treatment of cells with FK506 decreased Ca2+ content in thapsigargin-sensitive stores, suggesting that the partial depletion of the stores causes the inhibition of Ca2+ oscillations. Immunocytochemistry revealed the existence of cytoplasmic FKBP-like immunoreactivities. The expression of a 12-kDa FKBP was greater in subconfluent cells than in confluent cells as determined by Western blotting, suggesting that the 12-kDa FKBP may be one of the factors that regulates Ca2+ oscillations. Therefore, FK506 possesses an inhibitory action on the Ca2+ response via intracellular FKBP but not via calcineurin, which may result in modification of airway epithelial functions. PMID- 10362713 TI - Reactive oxygen intermediates stimulate interleukin-6 production in human bronchial epithelial cells. AB - Reactive oxygen intermediates (ROIs) play an important role in the initiation and progression of lung diseases. In this study, we investigated whether ROIs were involved in the induction of interleukin (IL)-6 in human bronchial epithelial cells. We exposed normal human bronchial epithelial cells as well as a human bronchial epithelial cell line, HS-24, to ROIs. We measured the amount of IL-6 in the culture supernatants using ELISA and the IL-6 mRNA levels using RT-PCR. Superoxide anions (O-2), but not hydrogen peroxide (H2O2), increased IL-6 production. To examine whether it is a cell type-specific mechanism of airway epithelial cells, the experiments were also performed in human lung fibroblasts, WI-38-40. In WI-38-40 cells, neither O-2 nor H2O2 increased IL-6 production. In contrast, tumor necrosis factor (TNF)-alpha (200 U/ml) induced IL-6 at the protein and mRNA levels in both airway epithelial cells and lung fibroblasts. This cytokine-induced IL-6 production was significantly suppressed by several antioxidants, including dimethyl sulfoxide (DMSO), in airway epithelial cells. In WI-38-40 cells, DMSO was not able to suppress IL-6 production induced by TNF alpha. Pretreatment with DMSO recovered the TNF-alpha-induced depletion of intracellular reduced glutathione in HS-24 cells. These findings indicate that oxidant stress specifically induces IL-6 production in human bronchial epithelial cells and that in these cells ROIs may be involved in IL-6 production after stimulation with cytokines such as TNF-alpha. Presumably, ROIs participate in the local immune response in lung diseases via IL-6 release from bronchial epithelial cells. PMID- 10362714 TI - Acute hypoxia increases alveolar macrophage tumor necrosis factor activity and alters NF-kappaB expression. AB - Alterations in alveolar macrophage (AM) function during sepsis-induced hypoxia may influence tumor necrosis factor (TNF) secretion and the progression of acute lung injury. Nuclear factor (NF)-kappaB is thought to regulate the expression of endotoxin [lipopolysaccharide (LPS)]-induced inflammatory cytokines such as TNF, and NF-kappaB may also be influenced by changes in O2 tension. It is thus proposed that acute changes in O2 tension surrounding AMs alter NF-kappaB activation and TNF secretion in these lung cells. AM-derived TNF secretion and NF kappaB expression were determined after acute hypoxic exposure of isolated Sprague-Dawley rat AMs. Adhered AMs (10(6)/ml) were incubated (37 degrees C at 5% CO2) for 2 h with LPS (Pseudomonas aeruginosa, 1 microgram/ml) in normoxia (21% O2-5% CO2) or hypoxia (1.8% O2-5% CO2). AM-derived TNF activity was measured with a TNF-specific cytotoxicity assay. Electrophoretic mobility shift and supershift assays were used to determine NF-kappaB activation and to identify NF-kappaB isoforms in AM extracts. In addition, mRNAs for selected AM proteins were determined with RNase protection assays. LPS-exposed AMs in hypoxia had higher levels of TNF (P < 0.05) and enhanced expression of NF-kappaB (P < 0.05); the predominant isoforms were p65 and c-Rel. Increased mRNA bands for TNF-alpha, interleukin-1alpha, and interleukin-1beta were also observed in the hypoxic AMs. These results suggest that acute hypoxia in the lung may induce enhanced NF kappaB activation in AMs, which may result in increased production and release of inflammatory cytokines such as TNF. PMID- 10362715 TI - SP-A 3'-UTR is involved in the glucocorticoid inhibition of human SP-A gene expression. AB - The synthetic glucocorticoid dexamethasone has a major inhibitory effect on human surfactant protein A1 (SP-A1) and SP-A2 gene expression that occurs at both the transcriptional and posttranscriptional levels. Toward the identification of cis acting elements that may be involved in the dexamethasone regulation of SP-A mRNA stability, chimeric chloramphenicol acetyltransferase (CAT) constructs that contained various portions of SP-A1 or SP-A2 cDNA in place of the native CAT 3' untranslated region (UTR) were transiently transfected into the lung adenocarcinoma cell line NCI-H441. CAT activity was reduced in NCI-H441 cells by exposure to 100 nM dexamethasone only for the chimeric CAT constructs that contained the SP-A 3'-UTR. Moreover, the inhibitory response seen with dexamethasone was greater for the 3'-UTR derived from the SP-A1 allele 6A3 than with the 3'-UTR derived from either the SP-A1 allele 6A2 or SP-A2 allele 1A0, indicating differential regulation between SP-A genes and/or alleles. PMID- 10362716 TI - NO causes perinatal pulmonary vasodilation through K+-channel activation and intracellular Ca2+ release. AB - Evidence suggests that nitric oxide (NO) causes perinatal pulmonary vasodilation through K+-channel activation. We hypothesized that this effect worked through cGMP-dependent kinase-mediated activation of Ca2+-activated K+ channel that requires release of intracellular Ca2+ from a ryanodine-sensitive store. We studied the effects of 1) K+-channel blockade with tetraethylammonium, 4 aminopyridine, a voltage-dependent K+-channel blocker, or glibenclamide, an ATP sensitive K+-channel blocker; 2) cyclic nucleotide-sensitive kinase blockade with either KT-5823, a guanylate-sensitive kinase blocker, or H-89, an adenylate sensitive kinase blocker; and 3) blockade of intracellular Ca2+ release with ryanodine on NO-induced pulmonary vasodilation in acutely prepared late-gestation fetal lambs. N-nitro-L-arginine, a competitive inhibitor of endothelium-derived NO synthase, was infused into the left pulmonary artery, and tracheotomy was placed. The animals were ventilated with 100% oxygen for 20 min, followed by ventilation with 100% oxygen and inhaled NO at 20 parts/million (ppm) for 20 min. This represents the control period. In separate protocols, the animals received an intrapulmonary infusion of the different blockers and were ventilated as above. Tetraethylammonium (n = 6 animals) and KT-5823 (n = 4 animals) attenuated the response, whereas ryanodine (n = 5 animals) blocked NO-induced perinatal pulmonary vasodilation. 4-Aminopyridine (n = 5 animals), glibenclamide (n = 5 animals), and H-89 (n = 4 animals) did not affect NO-induced pulmonary vasodilation. We conclude that NO causes perinatal pulmonary vasodilation through cGMP-dependent kinase-mediated activation of Ca2+-activated K+ channels and release of Ca2+ from ryanodine-sensitive stores. PMID- 10362717 TI - Respiratory epithelial cells demonstrate lactoferrin receptors that increase after metal exposure. AB - Human airway epithelial cells can increase expression of both lactoferrin and ferritin after exposure to catalytically active metal. These proteins transport and store metal, with coordination sites fully complexed, and therefore can diminish the oxidative stress. The intracellular transport of lactoferrin results in a transfer of complexed metal to ferritin, where it is stored in a less reactive form. This effort to control the injurious properties of metals would be facilitated by lactoferrin receptors (LfRs) on airway epithelial cells. We tested the hypotheses that 1) LfRs exist on respiratory epithelial cells and 2) exposure to both an air pollution particle, which has abundant concentrations of metals, and individual metal salts increase the expression of LfRs. Before exposure to either the particle or metals, incubation of BEAS-2B cells with varying concentrations of 125I-labeled lactoferrin demonstrated lactoferrin binding that was saturable. Measurement of 125I-lactoferrin binding after the inclusion of 100 micrograms/ml of oil fly ash in the incubation medium demonstrated increased binding within 5 min of exposure, which reached a maximal value at 45 min. Inclusion of 1.0 mM deferoxamine in the incubation of BEAS-2B cells with 100 micrograms/ml of oil fly ash decreased lactoferrin binding. Comparable to the particle, exposure of BEAS-2B cells to either 1.0 mM vanadyl sulfate or 1.0 mM iron (III) sulfate, but not to nickel sulfate, for 45 min elevated LfR activity. We conclude that LfRs on respiratory epithelial cells increased after exposure to metal. LfRs could participate in decreasing the oxidative stress presented to the lower respiratory tract by complexing catalytically active metals. PMID- 10362718 TI - Bleomycin stimulates lung epithelial cells to release neutrophil and monocyte chemotactic activities. AB - Although bleomycin, an antineoplastic drug, is used in the treatment of a variety of tumors, the mechanisms of bleomycin-induced lung injury and fibrosis are not fully elucidated. We postulated that bleomycin might stimulate A549 cells, a type II pneumocyte cell line, to release neutrophil and monocyte chemotactic activities (NCA and MCA, respectively). To test this hypothesis, A549 cell supernatant fluids were harvested and evaluated for NCA and MCA. A549 cell supernatant fluids showed NCA and MCA in response to bleomycin in a dose- and time-dependent manner (P < 0.05). Checkerboard analysis revealed that both NCA and MCA were predominantly chemotactic. Partial characterization of the released NCA and MCA showed that the activities were partially heat labile, trypsin digested, and predominantly ethyl acetate extractable. Lipoxygenase inhibitors and cycloheximide inhibited the release of chemotactic activities significantly. Molecular-sieve column chromatography revealed that the released activities were heterogeneous. However, low-molecular-weight activity was prominent. Leukotriene B4-receptor antagonist, anti-interleukin-8, anti-granulocyte colony-stimulating factor, and anti-monocyte chemoattractant protein-1 antibodies attenuated the chemotactic activities. Immunoreactive leukotriene B4 receptor, interleukin-8, granulocyte colony-stimulating factor, and monocyte chemoattractant protein-1 significantly increased in supernatant fluids in response to bleomycin. These data demonstrate that bleomycin stimulates type II epithelial cells to release chemotactic activities and plays a role in inflammatory cell recruitment into the lung. PMID- 10362719 TI - Cholinomimetic action of macrolide antibiotics on airway gland electrolyte secretion. AB - We investigated the acute effects of erythromycin (EM) and its derivatives on ionic currents in airway glands from feline tracheae. Therapeutic concentrations of EM or clarithromycin (CAM) attenuated the whole cell currents evoked by ACh in a competitive manner. The maximally stimulated inward Cl- currents were reduced to 54 and 83% and the outward K+ currents to 55 and 84% of control values by EM and CAM, respectively, whereas the responses induced by phenylephrine, norepinephrine, caffeine, or ionomycin were unaffected by EM, CAM, or EM523, a synthetic derivative of EM. K+ channels in excised outside-out patches were not influenced by macrolides. Although therapeutic concentrations of macrolides showed no effect on the baseline currents, high concentrations of macrolides alone evoked currents mimicking the ACh response, which were abolished completely by atropine. We concluded that macrolides act as a partial agonist on cholinergic receptors, resulting in a reduction of Cl- secretion at pharmacological doses of the agents, which may exhibit a pronounced effectiveness on hypertrophied and/or cholinergically sensitized submucosal glands in pathological airways. PMID- 10362720 TI - Segmental microvascular permeability in ischemia-reperfusion injury in rat lung. AB - Segmental microvascular permeabilities were measured using pre- and postalveolar vessel capillary filtration coefficient (Kfc) values (ml. min-1. cmH2O-1. 100 g 1) in isolated rat lungs subjected to ischemia-reperfusion (I/R). Total Kfc values measured in flowing and nonflowing lungs were highly correlated (r = 0.98, P < 0.0001). Kfc values were then measured in another group of lungs under no flow conditions when airway pressure was increased to 20 cmH2O and either the arterial or venous pressure was elevated to 7-8 cmH2O to measure the prealveolar and postalveolar Kfc values. Control total and postalveolar Kfc values were 0.0225 +/- 0.001 and 0.0219 +/- 0.001 ml. min-1. cmH2O-1. 100 g-1, respectively, and the prealveolar permeability was extremely small (0.00003 +/- 0.00005 ml. min 1. cmH2O-1. 100 g-1). Kfc values were again made in nonflowing lungs that had been subjected to 45 min of ischemia followed by 30 min of reperfusion. After I/R, the total membrane Kfc increased 10-fold to 0.2597 +/- 0.006 ml. min-1. cmH2O-1. 100 g-1, the prealveolar Kfc increased to 0.0677 +/- 0.003 ml. min-1. cmH2O-1. 100 g-1, and the postalveolar Kfc increased to 0.1354 +/- 0.008 ml. min 1. cmH2O-1. 100 g-1 (P < 0.05 for all I/R values). These data indicate that normal solvent microvascular permeability was predominantly postalveolar, and after I/R damage, the postalveolar (venular) permeability comprised 52% of the total, whereas the prealveolar and alveolar vessels comprised only 27 and 23%, respectively, of the total Kfc. PMID- 10362721 TI - Role of alveolar epithelial cell intercellular adhesion molecule-1 in host defense against Klebsiella pneumoniae. AB - Intercellular adhesion molecule-1 (ICAM-1) is expressed at high levels on type I alveolar epithelial cells (AEC) in the normal alveolar space. We postulate that AEC ICAM-1 enhances the antimicrobial activity of macrophages and neutrophils in the alveolar space. Wild-type and mutant mice deficient in ICAM-1 were inoculated intratracheally with Klebsiella pneumoniae. After 10 days, 43% of the ICAM-1 mutant mice had died compared with 14% of the wild-type controls (P = 0.003). Significantly more bacteria were isolated from lungs of ICAM-1 mutant mice than controls 24 h after inoculation (log colony-forming units 5.14 +/- 0.21 vs. 3.46 +/- 0. 16, P = 0.001). However, neutrophil recruitment to the lung was not different. In similar experiments in the rat, inhibition of alveolar ICAM-1 by intratracheal administration of antibody resulted in significantly impaired clearance of K. pneumoniae. The role of phagocyte interactions with AEC ICAM-1 for antimicrobial activity was investigated in vitro using primary cultures of rat AEC that express abundant ICAM-1. Alveolar macrophage phagocytosis and killing of K. pneumoniae were increased significantly in the presence of AEC; these effects were inhibited significantly (47.5 and 52%, respectively) when AEC ICAM-1 was blocked. Similarly, neutrophil phagocytic activity for K. pneumoniae in the presence of AEC in vitro was decreased when ICAM-1 on the AEC surface was blocked. Thus in the absence of ICAM-1, there is impaired ability to clear K. pneumoniae from the lungs, resulting in increased mortality. These studies indicate that AEC ICAM-1 plays an important role in host defense against K. pneumoniae by determining the antimicrobial activity of phagocytes within the lung. PMID- 10362722 TI - Expression of the Na+/H+ and Cl-/HCO-3 exchanger isoforms in proximal and distal human airways. AB - Recent studies have indicated the presence of Na+/H+ and Cl-/HCO-3 exchange activities in lung alveolar and tracheal tissues of various species. To date, the identity of the Na+/H+ (NHE) and Cl-/HCO-3 (AE) exchanger isoforms and their regional distribution in human airways are not known. Molecular species of the NHE and AE gene families and their relative abundance in the human airway regions were assessed utilizing RT-PCR and the RNase protection assay, respectively. Organ donor lung epithelia from various bronchial regions (small, medium, and large bronchi and trachea) were harvested for RNA extraction. Gene-specific primers for the human NHE and AE isoforms were utilized for RT-PCR. Our results demonstrated that NHE1, AE2, and brain AE3 isoforms were expressed in all regions of the human airways, whereas NHE2, NHE3, AE1, and cardiac AE3 were not detected. RNase protection studies for NHE1 and AE2, utilizing glyceraldehyde-3-phosphate dehydrogenase as an internal standard, demonstrated that there were regional differences in the NHE1 mRNA levels in human airways. In contrast, the levels of AE2 mRNA remained unchanged. Differential expression of these isoforms in the human airways may have functional significance related to the airway absorption and secretion of electrolytes. PMID- 10362723 TI - Silica-induced chemokine expression in alveolar type II cells is mediated by TNF alpha-induced oxidant stress. AB - We have shown previously that epithelial cells may contribute to the inflammatory response in the lung after exposure to crystalline silica through the production of and response to specific chemokines and cytokines. However, the exact cellular and molecular responses of epithelial cells to silica exposure remain unclear. We hypothesize that non-oxidant-mediated silica-cell interactions lead to the upregulation of tumor necrosis factor-alpha (TNF-alpha), whereby TNF-alpha induced generation of reactive oxygen species (ROS) leads to the activation of the monocyte chemotactic protein (MCP)-1 and macrophage inflammatory protein (MIP)-2 genes. Using a murine alveolar type II cell line, murine lung epithelial (MLE)-15, we measured the early changes in TNF-alpha, MCP-1, and MIP-2 mRNA species after exposure of the cells to 18 micrograms/cm2 silica (cristobalite) in combination with various antioxidants. Total mRNA was isolated and assayed using an RNase protection assay after 6 h of particle exposure. We found that extracellular GSH could completely attenuate the cristobalite-induced expression of MCP-1 and MIP-2 mRNAs, whereas TNF-alpha mRNA levels were unaltered. We also found using the oxidant-sensitive dye 6-carboxy-2', 7'-dichlorodihydrofluorescein diacetate di(acetoxymethyl ester) that treatment of MLE-15 cells with cristobalite and TNF-alpha (1 ng/ml) resulted in ROS production. This ROS production could be inhibited with extracellular GSH treatment, and in the case of cristobalite-induced ROS, inhibition was also achieved with an anti-TNF-alpha antibody. The results support the hypothesis that TNF-alpha mediates cristobalite induced MCP-1 and MIP-2 expression through the generation of ROS. PMID- 10362724 TI - Regulation of endothelial cell myosin light chain kinase by Rho, cortactin, and p60(src). AB - Inflammatory diseases of the lung are characterized by increases in vascular permeability and enhanced leukocyte infiltration, reflecting compromise of the endothelial cell (EC) barrier. We examined potential molecular mechanisms that underlie these alterations and assessed the effects of diperoxovanadate (DPV), a potent tyrosine kinase activator and phosphatase inhibitor, on EC contractile events. Confocal immunofluorescent microscopy confirmed dramatic increases in stress-fiber formation and colocalization of EC myosin light chain (MLC) kinase (MLCK) with the actin cytoskeleton, findings consistent with activation of the endothelial contractile apparatus. DPV produced significant time-dependent increases in MLC phosphorylation that were significantly attenuated but not abolished by EC MLCK inhibition with KT-5926. Pretreatment with the Rho GTPase inhibitory C3 exotoxin completely abolished DPV-induced MLC phosphorylation, consistent with Rho-mediated MLC phosphatase inhibition and novel regulation of EC MLCK activity. Immunoprecipitation of EC MLCK after DPV challenge revealed dramatic time-dependent tyrosine phosphorylation of the kinase in association with increased MLCK activity and a stable association of MLCK with the p85 actin binding protein cortactin and p60(src). Translocation of immunoreactive cortactin from the cytosol to the cytoskeleton was noted after DPV in concert with cortactin tyrosine phosphorylation. These studies indicate that DPV activates the endothelial contractile apparatus in a Rho GTPase-dependent fashion and suggests that p60(src)-induced tyrosine phosphorylation of MLCK and cortactin may be important features of contractile complex assembly. PMID- 10362725 TI - Brief 95% O2 exposure effects on surfactant protein and mRNA in rat alveolar and bronchiolar epithelium. AB - In acute lung injury, a disturbed surfactant system may impair gas exchange. Previous evaluations of hyperoxia effects on surfactant proteins (SPs) followed exposures >1-2 days. To evaluate the effects of brief exposure to hyperoxia on the SP system, we exposed adult male rats to 95% O2 or air for 12, 36, and 60 h. SP-A, -B, and -C mRNAs were analyzed by Northern blot and semiquantitative in situ hybridization (ISH). SP-A and -B were analyzed in whole lung homogenates, lung lavage fluid, and fixed tissue by semiquantitative immunohistochemistry (IHC). All SP mRNAs were diminished at 12 h and rose to or exceeded control by 60 h as determined by Northern blot and ISH. These effects were seen mainly in the intensity of ISH signal per cell in both type II and bronchiolar epithelial (Clara) cells and to a lesser extent on numbers of positively labeled cells. SP-B declined to 50% of control in lavage at 12 h, but no changes in total lung SP-A and -B were seen. The number of SP-A positively labeled cells did not change, but SP-A label intensity measured by IHC in type II cells showed parallel results to Northern blots and ISH. The response of SP-A in Clara cells was similar. SP-B immunolabeling intensity rose in both type II and Clara cells throughout the exposure. SP-C ISH intensity fell at 12 h and was increased to two times control by 60 h of hyperoxia. Sharp declines in SP expression occurred by 12 h of 95% O2 and may affect local alveolar stability. PMID- 10362726 TI - Mechanisms regulating cAMP-mediated growth of bovine neonatal pulmonary artery smooth muscle cells. AB - Neonatal pulmonary artery smooth muscle cells (PASMCs) exhibit enhanced growth capacity and increased growth responses to mitogenic stimuli compared with adult PASMCs. Because intracellular signals mediating enhanced growth responses in neonatal PASMCs are incompletely understood, we questioned whether 1) Gq agonists increase cAMP content and 2) increased cAMP is proproliferative. Endothelin-1 and angiotensin II increased both cAMP content and proliferation in neonatal but not in adult PASMCs. Inhibition of protein kinase C and protein kinase A activity nearly eliminated the endothelin-1- and angiotensin II-induced growth of neonatal PASMCs. Moreover, cAMP increased proliferation in neonatal but not in adult cells. Protein kinase C-stimulated adenylyl cyclase was expressed in both cell types, suggesting that insensitivity to stimulation of cAMP in adult cells was not due to decreased enzyme expression. Our data collectively indicate that protein kinase C stimulation of cAMP is a critical signal mediating proliferation of neonatal PASMCs that is absent in adult PASMCs and therefore may contribute to the unique proproliferative phenotype of these neonatal cells. PMID- 10362727 TI - Inhibition of gap junction communication in alveolar epithelial cells by 18alpha glycyrrhetinic acid. AB - Cultured alveolar epithelial cells exhibit gap junction intercellular communication (GJIC) and express regulated levels of connexin (Cx) 43 mRNA and protein. Newly synthesized radiolabeled Cx43 protein equilibrates with phosphorylated Cx43 isoforms; these species assemble to form both connexons and functional gap junction plaques. The saponin 18alpha-glycyrrhetinic acid (GA) rapidly and reversibly blocks GJIC at low concentrations (5 microM). Extended exposure to 18alpha-GA at higher concentrations causes inhibition of GJIC and time- and dose-dependent reductions in both Cx43 protein and mRNA expression. The latter toxic effects are paralleled by disassembly of gap junction plaques and are reversed less readily than acute effects on GJIC. These observations demonstrate 18alpha-GA-sensitive regulation of intercellular communication in epithelial cells from the mammalian lung and suggest a role for Cx43 expression and phosphorylation in acute and chronic regulation of GJIC between alveolar epithelial cells. PMID- 10362729 TI - Effects of a perfluorochemical emulsion on the fate of circulating Pseudomonas aeruginosa. AB - Because mononuclear phagocytes take up perfluorochemical emulsions (PFCE), we examined how prior treatment with PFCE affects the fate of circulating bacteria. Rats were preinjected with three daily intravenous injections of PFCE (2.0 ml/100 g) containing 12.5% (vol/vol) of a 4:1 mixture of F-dimethyl adamantane and F trimethylbicyclo-nonane, 2.5% (wt/vol) Pluronic F-68 as the emulsifying agent, and 3% (wt/vol) hydroxyethyl starch as the oncotic agent. Pseudomonas aeruginosa or Staphylococcus aureus were injected 4 h after the third PFCE injection. PFCE pretreatment decreased the rate and extent of vascular clearance of P. aeruginosa, with decreased uptake by the liver. Importantly, there were significant decreases in killing of P. aeruginosa in the liver, lungs, spleen, and kidneys of PFCE animals. PFCE did not alter the clearance of S. aureus from the circulation. However, hepatic uptake was reduced, with concomitant increases in lung and kidney uptake. Ultrastructure of Kupffer cells revealed PFCE inclusions and extensive vacuolization. These experiments demonstrate that the clearance kinetics and organ distribution of circulating P. aeruginosa and their subsequent killing are altered by PFCE. Diminished hepatic phagocyte function leads to a decrease in vascular clearance of circulating bacteria, increased uptake in other reticuloendothelial organs, and decreased bactericidal activity versus P. aeruginosa. PMID- 10362728 TI - A novel developmentally regulated gene in lung mesenchyme: homology to a tumor derived trypsin inhibitor. AB - We used differential display-PCR (DD-PCR) to identify glucocorticoid-inducible genes that regulate lung development in late gestation. DD-PCR, a method to screen for differentially expressed genes, is based on a comparison of mRNAs isolated from a subset of two or more cell populations by analysis of RT-PCR products on DNA-sequencing gels. We isolated cDNA probes representing mRNAs expressed in primary cultures of rat lung fibroblasts, but not in epithelial cells, on fetal day 20. A day 20 glucocorticoid-treated fibroblast cDNA library was screened with a single probe to isolate the 3.1-kb cDNA late-gestation lung 1 (LGL1; GenBank accession no. AF109674) encoding a deduced polypeptide of 188 amino acids. Northern analysis confirmed that LGL1 is expressed in human, rat, and mouse fetal lungs, induced by glucocorticoid, developmentally regulated in fibroblasts but not detectable in epithelium. In situ hybridization confirmed LGL1 expression in the mesenchyme, but not in the epithelium, of fetal rat lung, kidney, and gut. The predicted LGL1 gene product (lgl1) showed 81% homology to P25TI, a polypeptide trypsin inhibitor recently identified in human glioblastoma and neuroblastoma cells but not detected in normal human tissues. Both lgl1 and P25TI belong to the CRISP family of cysteine-rich extracellular proteins. Trypsin is produced by both normal bronchial epithelial and lung adenocarcinoma cells. Although additional studies will be necessary to clearly establish a functional role for lgl1, we propose that lgl1 has a role in normal lung development that is likely to be via regulation of extracellular matrix degradation. PMID- 10362731 TI - Renal endothelial and macula densa NOS: integrated response to changes in extracellular fluid volume. AB - If, only 20 years ago, anyone had postulated that the absence of nitric oxide gas (NO) would lead to severe hypertension and destruction of the vascular bed of the kidney within weeks, it is not unlikely that smiles of pity would have appeared on the faces of fellow researchers. By now, this has become common knowledge, and hundreds of reports have appeared on the regulation of vascular and renal function by nitric oxide. The amount of information complicates the design of a concept on how NO participates in control of extracellular fluid volume (ECFV) by the kidney. This review analyzes the function of endothelial and macula densa NO synthase (NOS) in the regulation of renal function. From this analysis, endothelial NOS (eNOS)-derived NO is considered a modulator of vascular responses and of renal autoregulation in particular. Increases in renal perfusion pressure and sodium loading will increase eNOS activity, resulting in vasodilatation and depression of tubuloglomerular feedback system responsiveness. Endothelium derived NO seems important to buffer minute-to-minute variations in perfusion pressure and rapid changes in ANG II activity. In contrast, macula densa NOS is proposed to drive adaptations to long-term changes in distal delivery and is considered a mediator of renin formation. Increases in perfusion pressure and distal delivery will depress the activity and expression of the enzyme that coincides with, and possibly mediates, diminished renin activity. Together, the opposite responses of eNOS and macula densa NOS-derived NO to changes in ECFV lead to an appropriate response to restore sodium balance. The concept that the two enzymes with different localizations in the kidney and in the cell are producing the same product, displaying contrasting responses to the same stimulus but nevertheless exhibiting an integrated response to perturbation of the most important regulated variable by the kidney, i.e., the ECFV, may be applicable to other tissues. PMID- 10362730 TI - Antisense oligonucleotides against the alpha-subunit of ENaC decrease lung epithelial cation-channel activity. AB - Amiloride-sensitive Na+ transport by lung epithelia plays a critical role in maintaining alveolar Na+ and water balance. It has been generally assumed that Na+ transport is mediated by the amiloride-sensitive epithelial Na+ channel (ENaC) because molecular biology studies have confirmed the presence of ENaC subunits alpha, beta, and gamma in lung epithelia. However, the predominant Na+ transporting channel reported from electrophysiological studies by most laboratories is a nonselective, high-conductance channel that is very different from the highly selective, low-conductance ENaC reported in other tissues. In our laboratory, single-channel recordings from apical membrane patches from rat alveolar type II (ATII) cells in primary culture reveal a nonselective cation channel with a conductance of 20.6 +/- 1.1 pS and an Na+-to-K+ selectivity of 0.97 +/- 0.07. This channel is inhibited by submicromolar concentrations of amiloride. Thus there is some question about the relationship between the gene product observed with single-channel methods and the cloned ENaC subunits. We have employed antisense oligonucleotide methods to block the synthesis of individual ENaC subunit proteins (alpha, beta, and gamma) and determined the effect of a reduction in the subunit expression on the density of the nonselective cation channel observed in apical membrane patches on ATII cells. Treatment of ATII cells with antisense oligonucleotides inhibited the production of each subunit protein; however, single-channel recordings showed that only the antisense oligonucleotide targeting the alpha-subunit resulted in a significant decrease in the density of nonselective cation channels. Inhibition of the beta- and gamma-subunit proteins alone or together did not cause any changes in the observed channel density. There were no changes in open probability or other channel characteristics. These results support the hypothesis that the alpha subunit of ENaC alone or in combination with some protein other than the beta- or gamma-subunit protein is the major component of lung alveolar epithelial cation channels. PMID- 10362732 TI - Effect of nitric oxide synthase inhibitors on short-term appetite and food intake in humans. AB - Animal studies suggest that nitric oxide (NO) may be a physiological regulator of appetite; NO synthase (NOS) inhibition suppresses food intake in rats, mice, and chickens. It is not known whether NO has any effect on appetite in humans. We have used NG-monomethyl-L-arginine (L-NMMA) and NG-nitro-L-arginine methyl ester (L-NAME), both competitive, nonselective inhibitors of NOS, in two separate studies to evaluate the role of NO in the short-term regulation of appetite in humans. In study I, 13 men (18-25 yr) underwent paired studies, in randomized, double-blind fashion, after an overnight fast. L-NMMA (4 mg. kg-1. h-1) or saline (0.9%) was infused intravenously at a rate of 40 ml/h for 1.5 h. In study II, eight men (18-26 yr) underwent three randomized, double-blind studies after an overnight fast. L-NAME (75 or 180 micrograms . kg-1. h-1) or saline (0.9%) was infused intravenously at a rate of 20 ml/h for 120 min. Hunger and fullness were measured using visual analog scales; blood pressure and heart rate were monitored, and 30 min before the end of the infusion, subjects were offered a cold buffet meal. Total caloric intake and the macronutrient composition of the meal were determined. Both L-NMMA (P = 0.052) and L-NAME (P < 0.05; both doses) decreased heart rate, L-NMMA increased diastolic blood pressure (P < 0.01), and L NAME increased systolic blood pressure (P = 0.052). Neither drug had any effect on caloric intake or sensations of hunger or fullness. Despite having significant effects on cardiovascular function in the doses used, neither L-NMMA nor L-NAME had any effect on feeding, suggesting that NO does not affect short-term appetite or food intake in humans. PMID- 10362733 TI - CNS origins of the sympathetic nervous system outflow to brown adipose tissue. AB - Brown adipose tissue (BAT) plays a critical role in cold- and diet-induced thermogenesis. Although BAT is densely innervated by the sympathetic nervous system (SNS), little is known about the central nervous system (CNS) origins of this innervation. The purpose of the present experiment was to determine the neuroanatomic chain of functionally connected neurons from the CNS to BAT. A transneuronal viral tract tracer, Bartha's K strain of the pseudorabies virus (PRV), was injected into the interscapular BAT of Siberian hamsters. The animals were killed 4 and 6 days postinjection, and the infected neurons were visualized by immunocytochemistry. PRV-infected neurons were found in the spinal cord, brain stem, midbrain, and forebrain. The intensity of labeled neurons in the forebrain varied from heavy infections in the medial preoptic area and paraventricular hypothalamic nucleus to few infections in the ventromedial hypothalamic nucleus, with moderate infections in the suprachiasmatic and lateral hypothalamic nuclei. These results define the SNS outflow from the brain to BAT for the first time in any species. PMID- 10362734 TI - Osmolality: a physiological long-term regulator of lumbar sympathetic nerve activity and arterial pressure. AB - Acute infusion of hypertonic fluid increases mean arterial pressure (MAP) in part by elevating nonrenal sympathetic activity. However, it is not known whether chronic, physiological increases in osmolality also increase sympathetic activity. To test this hypothesis, MAP, heart rate (HR), and lumbar sympathetic nerve activity (LSNA) were measured in conscious, 48-h water-deprived rats (WD) during a progressive reduction in osmolality produced by a 2-h systemic infusion (0.12 ml/min) of 5% dextrose in water (5DW). Water deprivation significantly increased osmolality (308 +/- 2 vs. 290 +/- 2 mosmol/kgH2O, P < 0.001), HR (453 +/- 7 vs. 421 +/- 10 beats/min, P < 0.05), and LSNA (63.5 +/- 1.8 vs. 51.9 +/- 3.8% baroreflex maximum, P < 0.01). Two hours of 5DW infusion reduced osmolality (-15 +/- 5 mosmol/kgH2O), LSNA (-23 +/- 3% baseline), and MAP (-10 +/- 1 mmHg). To evaluate the role of vasopressin in these changes, rats were pretreated with a V1-vasopressin receptor antagonist. The antagonist lowered MAP (-5 +/- 1 mmHg) and elevated HR (32 +/- 7 beats/min) and LSNA (11 +/- 3% baseline) in WD (P < 0. 05), but not in water-replete, rats. 5DW infusion had a similar cumulative effect on all variables in V1-blocked WD rats, but had no effect in water-replete rats. Infusion of the same volume of normal saline in WD rats did not change osmolality, LSNA or MAP. Together these data indicate that, in dehydrated rats, vasopressin supports MAP and suppresses LSNA and HR and that physiological changes in osmolality directly influence sympathetic activity and blood pressure independently of changes in vasopressin and blood volume. PMID- 10362735 TI - Vestigial respiratory burst activity in wound macrophages. AB - Macrophages from experimental wounds in rats were tested for their capacity to generate reactive oxygen intermediates. Measurements of superoxide and H2O2 release, O-2-dependent lucigenin chemiluminescence, oxygen consumption, hexose monophosphate shunt flux, and NADPH oxidase activity in cell lysates indicated, at best, the presence of a vestigial respiratory burst response in these cells. The inability of wound cells to release O-2 was not rekindled by priming with endotoxin or interferon-gamma in vivo or in vitro. NADPH oxidase activity in a cell-free system demonstrated that wound macrophage membranes, but not their cytosols, were capable of sustaining maximal rates of O-2 production when mixed with their corresponding counterparts from human neutrophils. Immune detection experiments showed wound macrophages to be particularly deficient in the cytosolic component of the NADPH oxidase p47-phox. Addition of recombinant p47 phox to the human neutrophil-cell membrane/wound macrophage cytosol cell-free oxidase assay, however, failed to support O-2 production. Present findings indicate an unexpected deficit of wound macrophages in their capacity to generate reactive oxygen intermediates. PMID- 10362736 TI - Brain adaptation to acute hyponatremia in young rats. AB - Brain swelling after acute hyponatremia in prepubescent rats, in contrast to adults, has recently been associated with an increase in brain sodium and a high mortality that could be prevented by preadministration of testosterone. To reexamine the effect of acute hyponatremia in young brain, we measured brain water and solute content in prepubescent rats after induction of hyponatremia over 4 h with water and arginine vasopressin. An 18% decrease in plasma sodium was associated with a 13% increase in brain water and a decrease in brain sodium and glutamate contents. No animals died. To assess the effect of sex hormones on brain adaptation, prepubescent rats were pretreated with estrogen or testosterone before acute hyponatremia. Brain sodium and potassium contents were significantly reduced in comparison to normonatremia in testosterone-pretreated but not estrogen-pretreated animals. However, there was no difference between estrogen pretreated and testosterone-pretreated groups in mortality or in brain contents of water, electrolytes, or major organic osmolytes. In conclusion, we found that brain adaptation to acute hyponatremia in prepubescent rats is similar to that observed in adults. PMID- 10362737 TI - Dietary salt intake alters cardiovascular responses evoked from the rostral ventrolateral medulla. AB - The present experiments examined whether in rats consuming diets with either high NaCl content (8%) or low Na+ content (0.01%) for 2 wk excitatory inputs to the rostral ventrolateral medulla (RVLM) would be altered. In chloralose-anesthetized rats, injection of glutamate into the RVLM elicited a pressor response that, compared with rats fed a control diet, was 50% larger in rats fed a diet containing 8% NaCl and was 25% smaller in rats fed a diet containing 0.01% Na+. Pressor responses produced by electrical stimulation of sciatic nerve afferents, as well as by microinjections into the RVLM of L-dihydroxyphenylalanine or carbachol, were all potentiated by high dietary salt intake and reduced by low dietary salt intake. Dietary salt intake had no effect on pressor responses produced by intravenous injection of phenylephrine, indicating that salt-related alterations in cardiovascular responses produced by central activation could not be accounted for by changes in peripheral vascular reactivity. The decrease in arterial pressure produced by injection of glutamate into the nucleus of the solitary tract was also potentiated by the high salt diet, suggesting that the sensitivity of central baroreceptor reflex pathways may be altered by dietary NaCl. These results indicate that the amount of NaCl consumed in the diet can change the sensitivity of RVLM sympathoexcitatory neurons, and this change in sensitivity is not restricted to any particular class of cell surface receptors. PMID- 10362738 TI - Cardiac- and noncardiac-related coherence between sympathetic drives to muscles of different human limbs. AB - Partial coherence analysis was used to evaluate the extent to which coherence between resting muscle sympathetic activity (MSA) in different pairs of limbs in humans is explained by the common baroreceptor input and by other noncardiac related factors. Multiunit MSA in two or three nerves, arterial blood pressure, and electrocardiogram were recorded simultaneously. Correlated MSA consisted of a sharp periodic component at the heart rate and a wideband component of relatively low power distributed between 0 and 2-2.5 Hz. Quantitative analysis revealed stronger coupling between MSAs in close limbs than in distant limbs (peak coherence leg-leg, 0. 94 +/- 0.03; arm-leg, 0.76 +/- 0.11). Furthermore, the wideband component, unaffected by partialization with circulatory signals, was significantly stronger between leg-leg (0.67 +/- 0.10) than between arm-leg pairs (0.29 +/- 0.10), i.e., noncardiac-related components explained 71% of leg-leg and 38% of arm-leg coherences at the frequency of the heart. Our results indicate that nonuniform relationship exists between resting sympathetic outflow to muscles in close and distant extremities which is, however, partially masked by the effect of the common rhythmic baroreceptor input. PMID- 10362739 TI - Capsaicin-sensitive fibers are required for the anorexic action of systemic but not central bombesin. AB - Bombesin (BN) suppresses food intake in rats whether given centrally or systemically. Although the brain BN-sensitive receptors are known to be essential for the anorexic effect of systemic BN, the mode of communication between the gut and the brain remains unclear. This study assessed whether the anorexic effect of systemic BN is mediated humorally or via neural circuits. Afferent neurons were lesioned using capsaicin (50 mg/kg sc) on postnatal day 2, and responses to BN were assessed during adulthood. Capsaicin treatment decreased body weight gain significantly from postnatal age 4-7 wk. Peripheral BN (4-16 micrograms/kg ip) dose dependently suppressed food intake in control animals. However, this effect was completely blocked in capsaicin-treated rats. In contrast to systemic effects, feeding-suppressant effects of centrally administered BN (0.01-0.5 microgram icv) were not affected by capsaicin treatment. This research suggests that peripheral BN communicates with the brain via a neuronal system(s) whose afferent arm is constituted of capsaicin-sensitive C and/or Adelta-fibers, whereas the efferent arm of this satiety- and/or anorexia-mediating circuitry is capsaicin resistant. PMID- 10362740 TI - Gut vagal afferent lesions increase meal size but do not block gastric preload induced feeding suppression. AB - Subdiaphragmatic vagal afferent (SVA) signals arising from gut sites may provide critical feedback for the control of food intake within a meal. To evaluate the role of SVAs in both spontaneous and scheduled meals, food intake was assessed in two paradigms in male Sprague-Dawley rats. In the first study, control (Con) rats (n = 6) and rats with subdiaphragmatic vagal deafferentation (SDA) (n = 7) had 12 h nightly access to Ensure liquid diet (1 kcal/ml). SDA rats had larger and fewer meals and maintained initial rapid rates of licking, yet total numbers of licks were unaffected. In the second study, Con (n = 8) and SDA (n = 7) rats had scheduled access to 12. 5% liquid glucose after overnight food deprivation. Glucose intake was assessed after 5-ml gastric preloads of 0.9% saline or glucose, peptone, and Intralipid solutions at three concentrations (0.5, 1, and 2 kcal/ml). Glucose and peptone preloads suppressed intake similarly in Con and SDA rats, whereas Intralipid was ineffective. These results suggest that meal-related SVA signals 1) are not critical in determining preload-induced feeding suppression after deprivation, yet 2) contribute to satiety during spontaneous meals. PMID- 10362741 TI - Functional evidence for subfornical organ-intrinsic conversion of angiotensin I to angiotensin II. AB - Using extracellular electrophysiological recording in an in vitro slice preparation, we investigated whether ANG I can be locally converted to the functionally active ANG II within the rat subfornical organ (SFO). ANG I and ANG II (10(-8)-10(-7) M) excited approximately 75% of all neurons tested with both peptides (n = 25); the remainder were insensitive. The increase in firing rate and the duration and the latency of the responses of identical neurons, superfused with equimolar concentrations of ANG I and ANG II, were not different. The threshold concentrations of the ANG I- and ANG II-induced excitations were both 10(-9) M. Inhibition of the angiotensin-converting enzyme by captopril (10( 4) M; n = 8) completely blocked the ANG I-induced excitation, a 10-fold lower dose was only effective in two of four neurons. The AT1-receptor antagonist losartan (10(-5) M; n = 6) abolished the excitation caused by ANG I and ANG II. Subcutaneous injections of equimolar doses of ANG I and ANG II (200 microliters; 2 x 10(-4) M) in water-sated rats similarly increased water intake by 2.4 +/- 0.5 (n = 16) and 2. 7 +/- 0.4 ml (n = 20) after 1 h, respectively. Control rats receiving saline drank 0.07 +/- 0.06 ml under these conditions. Pretreatment with a low dose of captopril (2.3 x 10(-3) M) 10 min before the injection of ANG I caused a water intake of 2.8 +/- 0.5 ml (n = 10), whereas a high dose of captopril (4.6 x 10(-1) M) suppressed the dipsogenic response of ANG I entirely (n = 11). These data provide direct functional evidence for an SFO-intrinsic renin-angiotensin system (RAS) and underline the importance of the SFO as a central nervous interface connecting the peripheral with the central RAS. PMID- 10362742 TI - Pressor reflex evoked by static muscle contraction: role of nitric oxide in the dorsal horn. AB - In this study, we tested the hypothesis that nitric oxide (NO) production in the dorsal horn is involved in producing the pressor reflex elicited by static contraction of skeletal muscle. Cats were anesthetized with alpha-chloralose (80 mg/kg) and urethane (100 mg/kg), and a laminectomy was performed. With the exception of the L7 dorsal root, the dorsal and ventral roots from L5 to S2 were sectioned on one side and static contraction of the ipsilateral triceps surae muscle was evoked by electrically stimulating the peripheral ends of the L7 and S1 ventral roots. Dialysis of the NO synthase inhibitor NG-nitro-L-arginine methyl ester (L-NAME; 50 mmol/l syringe concentration, based upon dose-response data) into the dorsal horn at L6 and S1 failed to attenuate the peak change in mean arterial pressure (MAP) evoked by static contraction (DeltaMAP in mmHg: 57 +/- 5 before and 50 +/- 6 after 2 h of L-NAME). However, this dialysis of L-NAME reduced the magnitude of the initial pressor response as the MAP at 10 s of the contraction fell from 27 +/- 4 to 17 +/- 4 mmHg. On the other hand, 2 h of L arginine dialysis (50 mmol/l) shifted the curve representing the time course of the pressor response upward and increased the peak pressor response to static contraction from 51 +/- 9 to 68 +/- 9 mmHg. A 2-h dialysis of D-NAME (50 mmol/l), the inactive enantiomer of L-NAME, had no effect on the time course or the peak pressor response (DeltaMAP in mmHg: 78 +/- 12 before and 72 +/- 15 after). These data suggest that NO production in the dorsal horn has a modulatory influence on the pressor reflex evoked by static contraction of skeletal muscle and that increasing the level of NO in the dorsal horn enhances the excitability of dorsal horn cells to muscle afferent input. PMID- 10362743 TI - Ontogeny of estrogen sulfatase activity in ovine fetal hypothalamus, hippocampus, and brain stem. AB - Ovine parturition is initiated by increases in fetal hypothalamus-pituitary adrenal (HPA) axis activity, which in turn increase placental estrogen biosynthesis and ultimately increase uterine contractility. In addition to the action in the uterus, estrogens augment fetal ACTH secretion. In late gestation, estrone sulfate is more abundant in fetal plasma than is unconjugated estrone. We studied hypothalamus, hippocampus, and brain stem tissue from fetal, neonatal, and adult sheep to test the hypothesis that the ovine brain contains estrogen sulfatase activity. We found that the activity in the hippocampus was significantly increased in late-gestation fetuses compared with both younger and older animals. No significant change in either hypothalamus or brain stem was revealed; however, the activity in all brain areas was high. Immunohistochemistry revealed the presence of estrogen sulfatase in the paraventricular nucleus of the hypothalamus, the nucleus of the solitary tract, and the rostral ventrolateral medulla. We conclude that ovine fetal hypothalamus, hippocampus, and brain stem contain estrogen sulfatase activity and that the activity in the hippocampus is developmentally regulated. PMID- 10362744 TI - Exposure to febrile temperature upregulates expression of pyrogenic cytokines in endotoxin-challenged mice. AB - Fever is a phylogenetically ancient response that is associated with improved survival in acute infections. In endothermic animals, fever is induced by a set of pyrogenic cytokines [tumor necrosis factor-alpha (TNF-alpha), interleukin (IL) 1, and IL-6] that are also essential for survival in acute infections. We studied the influence of core temperature on cytokine expression using an anesthetized mouse model in which core temperature was adjusted by immersion in water baths. We showed that raising core temperature from basal (36.5-37.5 degrees C) to febrile (39.5-40 degrees C) levels increased peak plasma TNF-alpha and IL-6 levels by 4.1- and 2. 7-fold, respectively, and changed the kinetics of IL-1beta expression in response to lipopolysaccharide challenge. TNF-alpha levels were increased predominantly in liver, IL-1beta levels were higher in lung, and IL-6 levels were widely increased in multiple organs in the warmer mice. This demonstrates that the thermal component of fever may directly contribute to shaping the host response by regulating the timing, magnitude, and tissue distribution of cytokine generation during the acute-phase response. PMID- 10362745 TI - Acetazolamide-induced cerebral and ocular vasodilation in humans is independent of nitric oxide. AB - Acetazolamide, a carbonic anhydrase inhibitor, is used orally in the treatment of primary and secondary open-angle glaucoma and induces ocular and cerebral vasodilation. Several in vitro studies have shown that carbonic anhydrase pharmacology and the L-arginine-nitric oxide (NO) pathway are closely related. We investigated the role of NO in acetazolamide-induced vasodilation on cerebral and ocular vessels in 12 healthy subjects in the presence or absence of NG-monomethyl L-arginine (L-NMMA), a NO synthase inhibitor, and in the presence or absence of L arginine, the precursor of NO. Acetazolamide was administered after pretreatment with either L-NMMA or placebo and either L-arginine or placebo. Pulsatile choroidal blood flow was assessed with laser interferometric measurement of fundus pulsation. In addition, mean blood flow velocity (MFV) in the middle cerebral artery (MCA) and ophthalmic artery (OA) was measured with Doppler sonography. Acetazolamide increased ocular fundus pulsation amplitude (FPA; +27%, P < 0.001) and MFV in the MCA (+38%, P < 0.001) and in the OA (+19%, P = 0.003). Administration of L-NMMA alone reduced FPA (-21%, P < 0.001) and MFV in the MCA ( 11%, P = 0. 030) but did not change MFV in the OA. All hemodynamic effects of L NMMA were reversed by L-arginine. However, neither L-NMMA nor L-arginine altered acetazolamide-induced changes in cerebral or ocular hemodynamic parameters. The present data indicate that acetazolamide-induced hemodynamic changes are not mediated by NO. Which mediators other than NO are involved in the hemodynamic effects as induced by carbonic anhydrase inhibitors remains to be elucidated. PMID- 10362746 TI - Effects of slow and rapid cooling on catecholamine concentration in arterial plasma and the skin. AB - Norepinephrine (NE) and epinephrine (Epi) concentrations in arterial plasma and in skin tissue were measured chromatographically before and after external cooling. Urethan-anesthetized rats were cooled either slowly (0.004-0.006 degrees C/s) or rapidly (0.03- 0.05 degrees C/s). Blood samples were drawn three times from each animal: 1) before cooling and at a rectal temperature decreased 2) by 0.5 degrees C and 3) by 3-4 degrees C. Skin samples were taken from controls and from rapidly or slowly cooled rats at a rectal temperature lowered by 0.5 degrees C. The resting mean values were 36.7 +/- 0.3 degrees C for rectal temperature, 0.62 +/- 0.079 and 1. 09 +/- 0.203 ng/ml for plasma NE and Epi, and 85.6 +/- 4.1 and 137.6 +/- 34.3 ng/g for skin NE and Epi. A decrease in rectal temperature by 0.5 degrees C at rapid cooling produced a 2.6-fold increase of NE and a 2.8-fold increase of Epi in plasma. Concomitantly, there was a significant decrease in skin NE concentration by 28% and Epi by 86%. At a rectal temperature decreased by 0.5 degrees C after slow cooling, plasma catecholamines did not change; at unaltered skin NE concentration, there was a reduction in skin Epi concentration (60%). When rectal temperature was lowered by 3-4 degrees C, the increase in plasma NE was virtually the same at both cooling rates and only plasma Epi increased more after deep rapid cooling than slow cooling. Thus the sympathoadrenal system may be differently activated depending on cooling rate. Rapid cooling, when the dynamic activity of the skin cold receptors is involved in the cold response, may provide conditions for an earlier activation of the sympathoadrenal system. This may evidence the functional significance of the dynamic activity of the skin cold receptors in the formation of the cold defense responses. PMID- 10362747 TI - Basolateral regulation of pHi in isolated snake renal proximal tubules in presence and absence of bicarbonate. AB - Intracellular pH (pHi) and its basolateral regulation were studied in isolated proximal-proximal and distal-proximal segments of garter snake (Thamnophis spp.) renal tubules with oil-filled lumens in HEPES-buffered and in HEPES-HCO-3 buffered media (pH 7.4 at 25 degrees C). pHi was measured with the pH-sensitive fluorescent dye 2',7'-bis(2-carboxyethyl)-5,6-carboxyfluorescein (BCECF) under resting conditions and in response to NH4Cl pulse. Resting pHi (approximately 7.1 7.2) and its response to and rate of recovery (dpHi/dt) from an NH4Cl pulse were not affected by the presence or absence of HCO-3 in either segment. Rate of recovery was depressed by Na+ removal in distal-proximal segments only and only in HEPES buffer. It was not affected by removal of Cl- or of both Na+ and Cl- or by reduction in membrane potential through addition of Ba2+ (5 mM) or high K+ (75 mM) in either segment in either HEPES or HEPES-HCO-3 buffer. The Na+/H+ exchange inhibitor ethylisopropylamiloride (EIPA) (100 microM) and the anion exchange inhibitor DIDS (100 microM) reduced dpHi/dt in the distal-proximal segments only and only in HEPES-HCO-3 buffer. The H+-ATPase inhibitor bafilomycin (1 microM), H+-K+-ATPase and K+/NH+4 exchange inhibitor Schering 28080 (10-100 microM), organic cation efflux inhibitor tetrapentylammonium (25 microM-20 mM), and K+ channel blocker tetraethylammonium (20 mM) had no effect on dpHi/dt in either segment. These data do not clearly support basolateral regulation of pHi in snake proximal renal tubules by commonly recognized Na+-dependent or Na+-independent acid or base transporters. PMID- 10362748 TI - Comparative analysis of NMR and NIRS measurements of intracellular PO2 in human skeletal muscle. AB - 1H NMR has detected both the deoxygenated proximal histidyl NdeltaH signals of myoglobin (deoxyMb) and deoxygenated Hb (deoxyHb) from human gastrocnemius muscle. Exercising the muscle or pressure cuffing the leg to reduce blood flow elicits the appearance of the deoxyMb signal, which increases in intensity as cellular PO2 decreases. The deoxyMb signal is detected with a 45-s time resolution and reaches a steady-state level within 5 min of pressure cuffing. Its desaturation kinetics match those observed in the near-infrared spectroscopy (NIRS) experiments, implying that the NIRS signals are actually monitoring Mb desaturation. That interpretation is consistent with the signal intensity and desaturation of the deoxyHb proximal histidyl NdeltaH signal from the beta subunit at 73 parts per million. The experimental results establish the feasibility and methodology to observe the deoxyMb and Hb signals in skeletal muscle, help clarify the origin of the NIRS signal, and set a stage for continuing study of O2 regulation in skeletal muscle. PMID- 10362749 TI - Kinetic profile of the rat CYP4A isoforms: arachidonic acid metabolism and isoform-specific inhibitors. AB - 20-Hydroxyeicosatetraenoic acid (HETE), the cytochrome P-450 (CYP) 4A omega hydroxylation product of arachidonic acid, has potent biological effects on renal tubular and vascular functions and on the control of arterial pressure. We have expressed high levels of the rat CYP4A1, -4A2, -4A3, and -4A8 cDNAs, using baculovirus and Sf 9 insect cells. Arachidonic acid omega- and omega-1 hydroxylations were catalyzed by three of the CYP4A isoforms; the highest catalytic efficiency of 947 nM-1. min-1 for CYP4A1 was followed by 72 and 22 nM 1. min-1 for CYP4A2 and CYP4A3, respectively. CYP4A2 and CYP4A3 exhibited an additional arachidonate 11,12-epoxidation activity, whereas CYP4A1 operated solely as an omega-hydroxylase. CYP4A8 did not catalyze arachidonic or linoleic acid but did have a detectable lauric acid omega-hydroxylation activity. The inhibitory activity of various acetylenic and olefinic fatty acid analogs revealed differences and indicated isoform-specific inhibition. These studies suggest that CYP4A1, despite its low expression in extrahepatic tissues, may constitute the major source of 20-HETE synthesis. Moreover, the ability of CYP4A2 and -4A3 to catalyze the formation of two opposing biologically active metabolites, 20-HETE and 11, 12-epoxyeicosatrienoic acid, may be of great significance to the regulation of vascular tone. PMID- 10362750 TI - Role of cholecystokinin in the anorexia produced by duodenal delivery of peptone in rats. AB - We used the cholecystokinin receptor antagonist devazepide to assess the importance of CCK in mediating the anorexia produced by 2-h duodenal infusions of peptone, a protein digest, at dark onset in nonfasted rats. Peptone alone (0.14 2.24 g/h) suppressed food intake dose dependently by 18-96%, with an approximate half-maximal dose of 1 g/h. Peptone-induced reductions in caloric ingestion were comparable to the caloric loads infused. Devazepide alone (30-1,000 microgram/kg) stimulated food intake dose dependently by 30-73%, with a minimal effective dose of 100 micrograms/kg. Devazepide appeared to reverse the anorexic response to peptone (1.1 g/h) dose dependently by 29-65%, with a minimal effective dose of 30 micrograms/kg. The magnitudes of these devazepide-induced effects were similar to, and in some cases were larger than, those produced when the same doses of devazepide were administered alone. Coadministration of devazepide (1,000 micrograms/kg) and a lower peptone dose (0.8 g/h) produced similar results. These results suggest that an essential CCK mechanism plays a significant role in mediating the satiety response to duodenal delivery of protein. PMID- 10362751 TI - Conditioned immunosuppression makes subtherapeutic cyclosporin effective via splenic innervation. AB - The present study investigated the mechanisms by which conditioned immunosuppression enhances the effectiveness of cyclosporin A (CsA) treatment in prolonging heart allograft survival. Dark Agouti rats that were administered subtherapeutic CsA (7 x 2 mg/kg on alternate days) rejected heart allografts at the same time as non-CsA-treated rats. The addition of a behavioral conditioning regimen (conditioned stimulus, saccharin; unconditioned stimulus, 20 mg/kg CsA) to the subtherapeutic CsA protocol produced a significant prolongation of graft survival, including long-term survival (>100 days) in 20% of the animals. Prior sympathetic denervation of the spleen completely blocked this effect. In nontransplanted rats both conditioning and CsA treatment reduce interleukin-2 and interferon (IFN)-gamma in the supernatant of proliferating splenocytes. Additionally, therapeutic CsA treatment decreased the number of IFN-gamma producing CD4(+) naive and memory T cells in the spleen. In contrast, behavioral conditioning increased that number. These data indicate that behavioral conditioning prolongs heart allograft survival by inhibiting the release of these cytokines in the spleen via sympathetic innervation, supplementing the inhibited cytokine production induced by CsA treatment. PMID- 10362752 TI - Role of nitric oxide in the early renal hemodynamic response after unilateral nephrectomy. AB - We evaluated the involvement of nitric oxide (NO) in the early hemodynamic response to uninephrectomy (UNX) in rats. Animals were uninephrectomized, and 48 h after removal of the kidney, the effect of infusing NG-nitro-L-arginine methyl ester (L-NAME) on renal function was studied. Glomeruli were isolated, and glomerular nitrite and cGMP productions were measured. In addition, endothelial constitutive NO synthase (NOS III) and inducible NO synthase (iNOS) were assessed by Western blot and by measuring the conversion of arginine to citrulline. UNX animals showed an increase in renal plasma flow that was inhibited by L-NAME in a higher proportion than in sham-operated (SO) animals. No differences were observed in systemic NO-dependent vascular tone, since mean arterial pressure showed similar increments in SO and UNX rats. Glomeruli from UNX animals showed an increase in glomerular nitrite production that was blunted by L-NAME addition. Also, cGMP levels were increased in glomeruli from UNX animals, and this increase was inhibited by L-NAME. Western blot analysis showed no differences in NOS III but a higher iNOS amount in glomeruli from UNX than in those from SO rats. No significant differences between UNX and SO rats were found in calcium-dependent NOS enzymatic activity in the renal cortex. However, calcium-independent enzymatic activity was markedly higher in the renal cortex of UNX than in those from SO animals. In conclusion, glomeruli from rats 48 h after UNX had a greater production of NO than those from SO animals. This increased glomerular NO production is based on an increase in the iNOS isoform. Increased glomerular NO synthesis seems to play a role in the decreased renal vascular resistance observed after unilateral nephrectomy in rats. PMID- 10362753 TI - Aging and spectral characteristics of the nonharmonic component of 24-h heart rate variability. AB - To examine whether heart rate variability (HRV) during daily life shows power law behavior independently of age and interindividual difference in the total power, log-log scaled coarse-graining spectra of the nonharmonic component of 24-h HRV were studied in 62 healthy men (age 21-79 yr). The spectra declined with increasing frequency in all subjects, but they appeared as broken lines slightly bending downward, particularly in young subjects with a large total power. Regression of the spectrum by a broken line with a single break point revealed that the spectral exponent (beta) was greater in the region below than above the break point (1.63 +/- 0. 23 vs. 0.96 +/- 0.21, P < 0.001). The break point frequency increased with age (r = 0.51, P < 0.001) and beta correlated with age negatively below the break point (r = 0.39) and positively above the break point (r = 0.70). The contribution to interindividual difference in total power was greater from the differences in the power spectral density at frequencies closer to both ends of the frequency axis and minimal from that at -3.25 log(Hz), suggesting hingelike movement of the spectral shape at this frequency with the difference in total power. These characteristics of the 24-h HRV spectrum were simulated by an artificial signal generated by adding two noises with different beta values. Given that the power law assumption is fundamental to the analysis of dynamics through the log-log scaled spectrum, our observations are substantial for physiological and clinical studies of the heartbeat dynamic during daily life and suggest that the nonharmonic component of HRV in normal subjects during daily life may include at least two 1/fbeta fluctuations that differ in dynamics and age dependency. PMID- 10362754 TI - Amiloride-sensitive sodium signals and salt appetite: multiple gustatory pathways. AB - In the rat, the ionic specificity of Na+ appetite is thought to rely on amiloride sensitive Na+ signals conveyed by the chorda tympani (CT) nerve. We evaluated whether robust Na+ appetite relies exclusively on CT-mediated amiloride-sensitive Na+ signals. Amiloride dramatically reduced sham drinking of NaCl (41.9 +/- 9.0 vs. 6.9 +/- 3.7 ml, 0.1 M NaCl without vs. with 100 microM amiloride), which resulted in intake that was not different from intake of a non-Na+ salt solution (8.8 +/- 2.3 ml, 0.15 M KCl). In addition, intake of 0.1 M NaCl in CT-transected (CTX) rats was reduced (35.8 +/- 13.3 vs. 8.67 +/- 3.4 ml, sham-operated vs. CTX rats), but the addition of amiloride (100 microM) further reduced intake in CTX rats (0.5 +/- 0.29 ml). These data support the idea that amiloride-sensitive Na+ channels are the critical gustatory substrate for Na+ identification during Na+ appetite in the rat. However, the data indicate that these amiloride-sensitive signals are not conveyed exclusively by the CT nerve but by an additional afferent pathway. PMID- 10362755 TI - Energy expenditure and balance during spaceflight on the space shuttle. AB - The objectives of this study were as follows: 1) to measure human energy expenditure (EE) during spaceflight on a shuttle mission by using the doubly labeled water (DLW) method; 2) to determine whether the astronauts were in negative energy balance during spaceflight; 3) to use the comparison of change in body fat as measured by the intake DLW EE, 18O dilution, and dual energy X-ray absorptiometry (DEXA) to validate the DLW method for spaceflight; and 4) to compare EE during spaceflight against that found with bed rest. Two experiments were conducted: a flight experiment (n = 4) on the 16-day 1996 life and microgravity sciences shuttle mission and a 6 degrees head-down tilt bed rest study with controlled dietary intake (n = 8). The bed rest study was designed to simulate the flight experiment and included exercise. Two EE determinations were done before flight (bed rest), during flight (bed rest), and after flight (recovery). Energy intake and N balance were monitored for the entire period. Results were that body weight, water, fat, and energy balance were unchanged with bed rest. For the flight experiment, decreases in weight (2.6 +/- 0.4 kg, P < 0.05) and N retention (-2. 37 +/- 0.45 g N/day, P < 0.05) were found. Dietary intake for the four astronauts was reduced in flight (3,025 +/- 180 vs. 1,943 +/- 179 kcal/day, P < 0.05). EE in flight was 3,320 +/- 155 kcal/day, resulting in a negative energy balance of 1,355 +/- 80 kcal/day (-15. 7 +/- 1.0 kcal. kg-1. day 1, P < 0.05). This corresponded to a loss of 2.1 +/- 0.4 kg body fat, which was within experimental error of the fat loss determined by 18O dilution (-1.4 +/- 0.5 kg) and DEXA (-2.4 +/- 0.4 kg). All three methods showed no change in body fat with bed rest. In conclusion, 1) the DLW method for measuring EE during spaceflight is valid, 2) the astronauts were in severe negative energy balance and oxidized body fat, and 3) in-flight energy (E) requirements can be predicted from the equation: E = 1.40 x resting metabolic rate + exercise. PMID- 10362756 TI - Regulation of P-450 4A activity in the glomerulus of the rat. AB - We recently reported that an enzyme of the cytochrome P-450 4A family is expressed in the glomerulus, but there is no evidence that 20 hydroxyeicosatetraenoic acid (20-HETE) can be produced by this tissue. The purpose of present study was to determine whether glomeruli isolated from the kidney of rats can produce 20-HETE and whether the production of this metabolite is regulated by nitric oxide (NO) and dietary salt intake. Isolated glomeruli produced 20-HETE, dihydroxyeicosatrienoic acids, and 12-hydroxyeicosatetraenoic acid (4.13 +/- 0.38, 4.20 +/- 0.38, and 2. 10 +/- 0.20 pmol. min-1. mg protein-1, respectively) when incubated with arachidonic acid (10 microM). The formation of 20-HETE was dependent on the availability of NADPH and the PO2 of the incubation medium. The formation of 20-HETE was inhibited by NO donors in a concentration dependent manner. The production of 20-HETE was greater in glomeruli isolated from the kidneys of rats fed a low-salt diet than in kidneys of rats fed a high salt diet (5.67 +/- 0.32 vs. 2.83 +/- 0.32 pmol. min-1. mg protein-1). Immunoblot experiments indicated that the expression of P-450 4A protein in glomeruli from the kidneys of rats fed a low-salt diet was sixfold higher than in kidneys of rats fed a high-salt diet. These results indicate that arachidonic acid is primarily metabolized to 20-HETE and dihydroxyeicosatrienoic acids in glomeruli and that glomerular P-450 activity is modulated by NO and dietary salt intake. PMID- 10362757 TI - Primary and adaptive changes of A-type K+ currents in sympathetic neurons from hypertensive rats. AB - The A-type K+ current (IA) of superior cervical ganglion neurons acutely isolated from spontaneously hypertensive (SHR) and age-matched Wistar-Kyoto (WKY) rats was compared under whole cell voltage clamp. Activation parameters were similar in each strain. Steady-state inactivation was shifted approximately -6 mV in SHR, where one-half inactivation occurred at -81 mV vs. -75 mV in WKY rats. The shift was not present in prehypertensive SHR but remained in adult enalapril-treated SHR and, therefore, may represent a primary alteration of IA properties. IA amplitudes evoked from physiological potentials were similar, despite inactivation of a greater fraction of the current in the SHR. Comparing maximal IA densities revealed that current density is elevated in the SHR, which compensates for the inactivation shift. Current density decreased with age in WKY neurons but did not significantly decline in SHR neurons unless hypertension was prevented with enalapril. Thus adult SHR neurons may retain a high IA density as an adaptive response to offset potential hyperexcitability resulting from the hyperpolarized IA inactivation. PMID- 10362758 TI - Identification of barosensitive neurons in the mediobasal forebrain using juxtacellular labeling. AB - Previous investigations suggest a possible role in cardiovascular regulation for neurons of the mediobasal forebrain. The present study was designed to determine the location and morphology of basal forebrain neurons that respond to acute changes in arterial blood pressure. Extracellular recordings of single units were done in alpha-chloralose- or urethan-anesthetized rats. The effect of cardiovascular pressor (phenylephrine, 1-2 microgram/kg iv) and depressor (sodium nitroprusside, 0.5-1 microgram/kg iv) events on the discharge rates of units was determined. Some of the neurons tested were subsequently filled with biocytin using the juxtacellular method. Brain sections were processed using the avidin biotin complex reaction to reveal a Golgi-like appearance of the neuron. Of 32 neurons located in the horizontal limb of the diagonal band of Broca (hDB), 13 (41%) were found to be excited by depressor events. Barosensitive biocytin labeled cells were located in all regions of the hDB and had small- to medium sized cell bodies with sparse and simple dendritic morphology. Only 2 of 47 neurons tested in the region of the olfactory tubercle, islands of Calleja (IC), and ventral pallidum responded to changes in arterial blood pressure. The results of the present investigation suggest a role in the regulation of cardiovascular function for neurons of the hDB. The findings also suggest that most neurons in the olfactory tubercle, including the IC complex, do not respond to acute changes in arterial blood pressure. PMID- 10362759 TI - Effect of dorsomedial hypothalamic nuclei knife cuts on ingestive behavior. AB - Previous findings show that rats with electrolytic or excitotoxic lesions in the dorsomedial hypothalamic nucleus (DMN) are hypophagic and hypodipsic and have reduced ponderal and linear growth but normal body composition. DMN-lesioned (DMNL) rats also show altered ingestive responses to naloxone. The present study investigated the intrahypothalamic nerve pathways involved in these DMNL effects and the response of the pathways to deprivation challenges by placing knife cuts posterior (Post), lateral (Lat), ventral (Vent), dorsal, or anterior to the DMN or by administering sham operations. One major finding was that rats with Post or Vent were hypophagic (P < 0. 05) and had reduced body weight but responded normally to deprivation challenges. Post and Lat groups were hypodipsic (P < 0. 05), but plasma Na+, K+, and osmolality and 24-h post-water-deprivation drinking responses were similar in all groups. Naloxone did not suppress the intake of Post rats. It appears that the hypophagia and the reduced body weight after DMNL involve fibers entering or leaving the DMN from ventral and posterior directions, and they may be part of an opioid feeding system. PMID- 10362760 TI - Visceral afferent activation-induced changes in sympathetic nerve activity and baroreflex sensitivity. AB - The following experiments were done to determine whether changes in baroreflex sensitivity evoked by cervical vagus nerve stimulation are due to sympathoexcitation mediated by the parabrachial nucleus. The relative contribution of cardiopulmonary and general gastric afferents within the cervical vagus nerve to the depression in baroreflex sensitivity are also investigated. Male Sprague-Dawley rats anesthetized with thiobutabarbital sodium (50 mg/kg) were instrumented to measure blood pressure and heart rate or for the continuous monitoring of renal sympathetic nerve activity. Baroreflex sensitivity was measured using bolus injections of phenylephrine. Electrical stimulation of the cervical vagus (with or without the aortic depressor nerve) or the abdominal vagus nerve produced a significant increase in renal nerve activity and a decrease in baroreflex sensitivity. Both of these effects were blocked after the microinjection of lidocaine into the parabrachial nucleus before nerve stimulation. Therefore, we conclude that an increase in the activity of cardiac, pulmonary, or general gastric afferents mediated the increased sympathetic output and decreased baroreflex sensitivity via a pathway involving the parabrachial nucleus. PMID- 10362762 TI - Can gender differences during exercise-heat stress be assessed by the physiological strain index? AB - A physiological strain index (PSI) based on rectal temperature (Tre) and heart rate (HR) was recently suggested to evaluate exercise-heat stress. The purpose of this study was to evaluate PSI for gender differences under various combinations of exercise intensity and climate. Two groups of eight men each were formed according to maximal rate of O2 consumption (VO2 max). The first group of men (M) was matched to a group of nine women (W) with similar (P > 0.001) VO2 max (46.1 +/- 2.0 and 43.6 +/- 2.9 ml. kg-1. min-1, respectively). The second group of men (MF) was significantly (P < 0. 001) more fit than M or W with VO2 max of 59.1 +/- 1.8 ml. kg-1. min-1. Subjects completed a matrix of nine experimental combinations consisting of three different exercise intensities for 60 min [low, moderate, and high (300, 500, and 650 W, respectively)] each at three climates (comfortable, hot wet, and hot dry [20 degrees C 50% relative humidity (RH), 35 degrees C 70% RH, and 40 degrees C 35% RH, respectively]). No significant differences (P > 0.05) were found between matched genders (M and W) at the same exposure for sweat rate, relative VO2 max (%VO2 max), and PSI. However, MF had significantly (P < 0.05) lower strain than M and W as reflected by %VO2 max and PSI. In summary, PSI applicability was extended for exercise-heat stress and gender. This index continues to show potential for wide acceptance and application. PMID- 10362761 TI - Neonatal rabbit proximal tubule basolateral membrane Na+/H+ antiporter and Cl /base exchange. AB - The present in vitro microperfusion study examined the maturation of Na+/H+ antiporter and Cl-/base exchanger on the basolateral membrane of rabbit superficial proximal straight tubules (PST). Intracellular pH (pHi) was measured with the pH-sensitive fluorescent dye 2', 7'-bis(2-carboxyethyl)-5(6) carboxyfluorescein in neonatal and adult superficial PST. Na+/H+ antiporter activity was examined after basolateral Na+ addition in tubules initially perfused and bathed without Na+. Neonatal Na+/H+ antiporter activity was approximately 40% that of adult segment (9.7 +/- 1.5 vs. 23.7 +/- 3.2 pmol. mm-1. min-1; P < 0.001). The effect of bath Cl- removal on pHi was used to assess the rates of basolateral Cl-/base exchange. In both neonatal and adult PST, the Cl /base exchange activity was significantly higher in the presence of 25 mM HCO-3 than in the absence of HCO-3 and was inhibited by cyanide and acetazolamide, consistent with Cl-/HCO-3 exchange. The proton flux rates in the presence of bicarbonate in neonatal and adult tubules were 14.1 +/- 3.6 and 19.5 +/- 3.5 pmol. mm-1min-1, respectively (P = NS), consistent with a mature rate of Cl-/HCO 3 exchanger activity in neonatal tubules. Basolateral Cl-/base exchange activity in the absence of CO2 and HCO-3, with luminal and bath cyanide and acetazolamide, was greater in adult than in neonatal PST and inhibited by bath DIDS consistent with a maturational increase in Cl-/OH- exchange. We have previously shown that the rates of the apical membrane Na+/H+ antiporter and Cl-/base exchanger were approximately fivefold lower in neonatal compared with adult rabbit superficial PST. These data demonstrate that neonatal PST basolateral membrane Na+/H+ antiporter and Cl-/base exchanger activities are relatively more mature than the Na+/H+ antiporter and Cl-/base exchangers on the apical membrane. PMID- 10362763 TI - Metabolic and cardiorespiratory responses to hypoxia in fetal sheep: adenosine receptor blockade. AB - 8-Phenyltheophylline (PT), a potent and specific inhibitor of adenosine receptors, was infused intra-arterially into unanesthetized fetal sheep to determine the role of adenosine in hypoxic inhibition of fetal breathing. PT in normoxic fetuses increased heart rate and the incidence of low-voltage electrocortical activity, rapid eye movements (REM), and breathing. Mean breath amplitude increased by 44%. Hypoxia (preductal arterial PO2 = 14 Torr) induced a metabolic acidemia, a transient bradycardia, and hypertension while virtually eliminating REM and breathing. PT administration during hypoxia enhanced the metabolic acidemia, blocked the bradycardia and hypertension, increased the incidence of REM and breathing, and elevated mean breath amplitude. The results indicate that 1) adenosine is involved in fetal glycolytic and cardiovascular responses to hypoxia, 2) activation of central adenosine receptors mediates about one-half the inhibitory effects of hypoxia on REM and breathing, and 3) the depression of breathing may critically depend on a hypoxia-induced reduction in phasic REM sleep. PMID- 10362764 TI - Nuclear factor-kappaB-like activity increases in murine cerebral cortex after sleep deprivation. AB - Several well-defined sleep regulatory substances, e.g., interleukin-1beta, activate the heterodimeric transcription factor nuclear factor-kappaB (NF kappaB). Several substances that inhibit sleep, e.g., interleukin-4, inhibit NF kappaB activation. NF-kappaB activation promotes production of several additional substances thought to be involved in sleep regulation, e.g., nitric oxide. We investigated, therefore, whether there are diurnal rhythms of NF-kappaB activation in brain and changes in the activation after sleep deprivation. Mice were kept on a 12:12-h light-dark cycle. In one experiment, groups of mice were killed every 3 h across the 24-h cycle. In another experiment, mice were killed at 1500 after 6 h of sleep deprivation, and a group of control mice were killed at the same time. Nuclear proteins were extracted from each brain tissue sample, and NF-kappaB-like activity was determined with an electrophoretic mobility shift assay. In cerebral cortex, but not other areas of brain, there was a diurnal rhythm in NF-kappaB-like activation; highest levels were found during the light period. NF-kappaB-like activation was higher in cerebral cortex after sleep deprivation compared with values obtained from control mice. The results are consistent with the hypothesis that sleep regulation involves multiple gene events, some of which include enhanced production of sleep regulatory substances, the actions of which involve NF-kappaB activation. PMID- 10362765 TI - Threshold for efferent bladder nerve firing in the rat. AB - In this study, the mechanism involved in the initiation of voiding was investigated. Bladder pressure and bladder and urethral nerve activity were recorded in the anesthetized rat. Bladder nerve activity was resolved into afferent and efferent activity by means of a theoretical model. The beginning of an active bladder contraction was defined as the onset of bladder efferent firing at a certain time (t0). From t0 onward, bladder efferent activity increased linearly during deltat seconds (rise time) to a maximum. The pressure at t0 was 1.0 +/- 0.4 kPa, the afferent nerve activity at t0 was 2.0 +/- 0.6 microV (53 +/- 15% of maximum total nerve activity), and deltat was 11 +/- 13 s. Between contractions the afferent activity at t0 was never exceeded. Urethral afferent nerve activity started at bladder pressures of 2.1 +/- 1.1 kPa. Therefore, we concluded that urethral afferent nerve activity does not play a role in the initiation of bladder contractions; voiding contractions presumably are initiated by bladder afferent nerve activity exceeding a certain threshold. PMID- 10362766 TI - Role of glucocorticoids in the maturation of renal cortical Na+-K+-ATPase during fetal life in sheep. AB - Glucocorticoid levels increase greatly at the time of birth in humans and sheep, coinciding with an increased ability of the kidney to reabsorb sodium. Cortisol induces proximal tubule apical membrane Na+/H+ exchanger maturation in near-term fetal sheep. Proximal tubule salt transport is ultimately dependent on Na+ pump activity, so we studied the effects of cortisol treatment on renal cortical Na+ K+-ATPase. We first looked at six 140 day gestation fetal sheep (term is 145) and compared their renal cortical Na+-K+-ATPase to that of six 1-day-old lambs. Na+ K+-ATPase activity increased 80% after birth. Then nine pairs of twin fetal sheep were chronically instrumented at 127 days gestation. After 72 h recovery, one twin was given a 48-h continuous intraperitoneal infusion of cortisol. Both twins were then killed, and their renal cortices were studied. Na+-K+-ATPase activity increased 122% with cortisol treatment; activity equaled that of 1-day-old lambs. Protein abundance of the alpha1-subunit of the Na+-K+-ATPase increased 19%; the beta1-subunit increased 39% with cortisol treatment. mRNA abundance of the alpha1 subunit increased 58%; the beta1-subunit increased 72%. These results indicate that cortisol matures Na+-K+-ATPase activity. PMID- 10362767 TI - Cold face test demonstrates parasympathetic cardiac dysfunction in familial dysautonomia. AB - In familial dysautonomia (FD), i.e., Riley-Day syndrome, parasympathetic dysfunction has not been sufficiently evaluated. The cold face test is a noninvasive method of activating trigeminal brain stem cardiovagal and sympathetic pathways and can be performed in patients with limited cooperation. We performed cold face tests in 11 FD patients and 15 controls. For 60 s, cold compresses (0-1 degrees C) were applied to the cheeks and forehead while we monitored heart rate, respiration, beat-to-beat radial artery blood pressure, and laser-Doppler skin blood flow at the first toe pulp. From these measurements heart rate variability parameters were calculated: root mean square of successive differences (RMSSD), coefficient of variation (CV), low- and high-frequency (LF and HF, respectively) power spectra of the electrocardiogram, and the LF transfer function gain between blood pressure and heart rate. All patients perceived cold stimulation and acknowledged discomfort. In controls, heart rate and skin blood flow decreased significantly during cold face test; in patients, both parameters decreased only briefly and not significantly. In controls, blood pressure, RMSSD, CV, and heart rate HF-power spectra increased but remained unchanged in patients. Respiration, as well as heart rate LF power spectra, did not change in either group. In controls, LF transfer function gain between blood pressure and heart rate indicated that bradycardia was not secondary to blood pressure increase. We conclude that the cold face test demonstrated that patients with FD have a reduced cardiac parasympathetic response, which implies efferent parasympathetic dysfunction. PMID- 10362768 TI - Blood-borne, albumin-bound prostaglandin E2 may be involved in fever. AB - Although the involvement of blood-borne PGE2 in fever has been hypothesized by several authors and has substantial experimental support, the current literature often rejects this hypothesis because several attempts to induce fever by a peripheral PGE2 failed. However, it is usually ignored that the amphipathic molecules of PGE2 are readily self-associating and that such an aggregation could have prevented the peripherally administered PGE2 (free form) from expressing its pyrogenic activity, thus leading to false negative results. To ensure disaggregation of PGE2, we prepared its complex within a carrier protein, human serum albumin (HSA). HSA was purified with activated charcoal and polymixin B polyacrylamide gel and incubated with PGE2 for 1 h at 37 degrees C. Adult Chinchilla rabbits were injected intravenously with PGE2-HSA complex in either the higher (75 micrograms/kg PGE2:30 mg/kg HSA) or the lower (15 micrograms/kg:6 mg/kg) dose, and the rectal temperature (Tr) was measured. In the controls, either the ligand alone or the carrier alone was administered. At the higher dose, neither free PGE2 nor albumin alone was pyrogenic, whereas the PGE2-HSA complex produced a fever characterized by a short latency (<10 min) and a maximal Tr rise of 0.7 +/- 0.2 degrees C. At the lower dose, none of the substances affected the Tr. This study demonstrates a marked pyrogenic activity of the intravenous PGE2-HSA, but not of the free PGE2. Administration of a preformed complex may be more physiologically relevant than administration of the free ligand because of the ligand's disaggregation, protection from enzymatic degradation, and facilitated delivery to targets. Our study supports the hypothesis that peripheral PGE2 is involved in fever genesis. PMID- 10362769 TI - H+-K+-ATPases: regulation and role in pathophysiological states. AB - Molecular cloning experiments have identified the existence of two H+-K+-ATPases (HKAs), colonic and gastric. Recent functional and molecular studies indicate the presence of both transporters in the kidney, which are presumed to mediate the exchange of intracellular H+ for extracellular K+. On the basis of these studies, a picture is evolving that indicates differential regulation of HKAs at the molecular level in acid-base and electrolyte disorders. Of the two transporters, gastric HKA is expressed constitutively along the length of the collecting duct and is responsible for H+ secretion and K+ reabsorption under normal conditions and may be stimulated with acid-base perturbations and/or K+ depletion. This regulation may be species specific. To date there are no data to indicate that the colonic HKA (HKAc) plays a role in H+ secretion or K+ reabsorption under normal conditions. However, HKAc shows adaptive regulation in pathophysiological conditions such as K+ depletion, NaCl deficiency, and proximal renal tubular acidosis, suggesting an important role for this exchanger in potassium, HCO-3, and sodium (or chloride) reabsorption in disease states. The purpose of this review is to summarize recent functional and molecular studies on the regulation of HKAs in physiological and pathophysiological states. Possible signals responsible for regulation of HKAs in these conditions will be discussed. Furthermore, the role of these transporters in acid-base and electrolyte homeostasis will be evaluated in the context of genetically altered animals deficient in HKAc. PMID- 10362770 TI - The nongastric H+-K+-ATPases: molecular and functional properties. AB - The Na-K/H-K-ATPase gene family is divided in three subgroups including the Na-K ATPases, mainly involved in whole body and cellular ion homeostasis, the gastric H-K-ATPase involved in gastric fluid acidification, and the newly described nongastric H-K-ATPases for which the identification of physiological roles is still in its infancy. The first member of this last subfamily was first identified in 1992, rapidly followed by the molecular cloning of several other members. The relationship between each member remains unclear. The functional properties of these H-K-ATPases have been studied after their ex vivo expression in various functional expression systems, including the Xenopus laevis oocyte, the insect Sf9 cell line, and the human HEK 293 cells. All these H-K-ATPase alpha subunits appear to encode H-K-ATPases when exogenously expressed in such expression systems. Recent data suggest that these H-K-ATPases could also transport Na+ in exchange for K+, revealing a complex cation transport selectivity. Moreover, they display a unique pharmacological profile compared with the canonical Na-K-ATPases or the gastric H-K-ATPase. In addition to their molecular and functional characterizations, a major goal is to correlate the molecular expression of these cloned H-K-ATPases with the native K-ATPases activities described in vivo. This appears to be more complex than anticipated. The discrepancies between the functional data obtained by exogenous expression of the nongastric H-K-ATPases and the physiological data obtained in native organs could have several explanations as discussed in the present review. Extensive studies will be required in the future to better understand the physiological role of these H-K-ATPases, especially in disease processes including ionic or acid-base disorders. PMID- 10362771 TI - Developmental expression of ROMK in rat kidney. AB - The apical secretory K+ (SK) channel in the principal cell represents the rate limiting step for K+ secretion across the cortical collecting duct (CCD). Patch clamp analysis of maturing rabbit principal cells identifies an increase in number of conducting SK channels after the 2nd week of life [L. M. Satlin and L. G. Palmer. Am. J. Physiol. 272 (Renal Physiol. 41): F397-F404, 1997], approximately 1 wk after an increase in activity of the amiloride-sensitive epithelial Na+ channel (ENaC) is detected. To correlate the postnatal increase in channel activity with developmental expression of ROMK, the molecular correlate of the SK channel, we used gene-specific probes to show a developmental increase in abundance of renal ROMK mRNA and a ROMK-specific antibody to examine the nephron distribution, localization, and abundance of this protein in developing rat kidney. Using antibodies directed against aquaporin-3 (AQP-3) and Tamm Horsfall protein (THP), we confirmed that ROMK was expressed along the apical membranes of principal cells and thick ascending limbs of Henle (TALH) in adult kidney. Within the midcortex of the neonatal kidney, ROMK-positive segments revealed weak coincident staining with the TALH-specific antibody but did not colabel with an antibody directed against distal and connecting tubule (CNT) specific kallikrein or the lectin Dolichos biflorus agglutinin (DBA), which labels proximal tubules and collecting ducts. In inner cortex and outer medulla of kidneys from 1-wk-old animals, ROMK protein was identified in medullary TALH (MTALH) and DBA-positive collecting ducts. By 3 wk of age, coincident ROMK and DBA expression was detected in midcortical and outer cortical CNTs and CCDs. Immunoblot analysis of plasma membrane-enriched fractions of maturing rat kidney revealed a developmental increase in a approximately 40-kDa band, the expected size for ROMK. Immunolocalization of alpha-ENaC showed apical staining of a majority of cells in distal nephron segments after the 1st week of postnatal life. The beta- and gamma-ENaC subunit expression was routinely detected in a mostly cytoplasmic distribution immediately after birth, albeit in low abundance; gamma-ENaC showed some apical polarization. These results suggest that the postnatal increases in a principal cell apical SK and Na+ channel activity are mediated, at least in part, by increases in abundance of ROMK message and protein and ENaC subunit proteins. PMID- 10362772 TI - Partial ATP depletion induces Fas- and caspase-mediated apoptosis in MDCK cells. AB - Brief periods of in vitro hypoxia/ischemia induce apoptosis of cultured renal epithelial cells, but the underlying mechanisms remain unknown. We show that partial ATP depletion (approximately 10-65% of control) results in a duration dependent induction of apoptosis in Madin-Darby canine kidney (MDCK) cells, as evidenced by internucleosomal DNA cleavage (DNA laddering and in situ nick end labeling), morphological changes (cell shrinkage), and plasma membrane alterations (externalization of phosphatidylserine). The ATP-depleted cells display a significant upregulation of Fas, Fas ligand, and the Fas-associating protein with death domain (FADD). Exogenous application of stimulatory Fas monoclonal antibodies also induces apoptosis in nonischemic MDCK cells, indicating that they retain Fas-dependent pathways of programmed cell death. Furthermore, cleavage of poly(ADP)ribose polymerase (PARP) is evident after ATP depletion, indicating activation of caspases. Indeed, the apoptotic cells display a significant increase in caspase-8 (FLICE) activity. Finally, apoptosis induced by ATP depletion is ameliorated by pretreatment with inhibitors of caspase-8 (IETD), caspase-1 (YVAD), or caspase-3 (DEVD) but is not affected by inhibitors of serine proteases (TPCK). Our results indicate that partial ATP depletion of MDCK cells results in apoptosis and that Fas- and caspase-mediated pathways may play a critical role. PMID- 10362773 TI - Pathways for HCO-3 exit across the basolateral membrane in rat thick limbs. AB - We studied the pathways for HCO-3 transport in basolateral membrane vesicles (BLMV) purified from rat medullary thick ascending limbs (MTAL). An inward HCO-3 gradient in the presence of an inside-positive potential stimulated the rate of 22Na uptake minimally and did not induce a 22Na overshoot, arguing against the presence of electrogenic Na+-HCO-3 cotransport in these membranes. An inside-acid pH gradient stimulated to the same degree uptake of 86Rb+ (a K+ analog) with or without HCO-3. Conversely, applying an outward K+ gradient caused a modest intracellular pH (pHi) decrease of approximately 0.38 pH units/min, as monitored by quenching of carboxyfluorescein; its rate was unaffected by HCO-3, indicating the absence of appreciable K+-HCO-3 cotransport. On the other hand, imposing an inward Cl- gradient in the presence of HCO-3 caused a marked pHi decrease of approximately 1.68 pH units/min; its rate was inhibited by a stilbene derivative. Finally, we could not demonstrate the presence of a HCO-3/lactate exchanger in BLMV. In conclusion, the presence of significant Na+-, K+-, or lactate-linked HCO 3 transport could not be demonstrated. These and other data suggest that basolateral Cl-/HCO-3 exchange could be the major pathway for HCO-3 transport in the MTAL. PMID- 10362774 TI - D-Serine is reabsorbed in rat renal pars recta. AB - D-Serine normally contributes up to 3% to total plasma serine and up to 23% in chronic renal failure. D-Serine is metabolized by tubular D-amino acid oxidase (D AAO), and high D-serine plasma levels are nephrotoxic; both events are localized in the straight part of the proximal tubule. We therefore investigated if and how D-serine is reabsorbed there. We microinfused 14C-labeled D- or -L-serine + [3H]inulin into early proximal (EP), late proximal (LP), or early distal (ED) tubule sections of superficial nephrons and into long loops of Henle (LLH) of rats in vivo and in situ. The fractional reabsorption (FR) of the 14C label was determined from the 14C:3H ratio in the final urine. At 0.36 mM, FR of D [14C]serine was 86% (EP), 90% (LP), and approximately 0 (ED, LLH). FR of D-serine could be saturated and inhibited by L-serine (and vice versa). D-methionine, but not D-glutamate or D-arginine, blocked FR of D-serine (LP). We conlude that filtered D-serine is able to enter the pars recta cells, thereby getting access to D-AAO. The uptake carrier has a very low stereospecificity and is, therefore, different from that in the proximal convolution. The colocalization of exclusive reabsorption and metabolism makes the pars recta the tubule site for the recycling of the carbon structure of D-amino acids and, at the same time, the target of D-serine nephrotoxicity. PMID- 10362775 TI - Localization of an organic anion transporter-GFP fusion construct (rROAT1-GFP) in intact proximal tubules. AB - The organic anion transporter, rROAT1, is a dicarboxylate/organic anion exchanger, a function associated with the basolateral membrane in rat proximal tubule. To directly establish the subcellular localization of rROAT1 in renal epithelia, we made a rROAT1-green fluorescent protein (GFP) fusion construct (rROAT1-GFP). Plasma membrane-associated fluorescence was observed in rROAT1-GFP expressing Xenopus oocytes examined by confocal microscopy. Uptake of 3H-labeled p-aminohippurate (PAH) increased 2. 5-fold in rROAT1-GFP-expressing Xenopus oocytes, and this increase was abolished by 1 mM probenecid. Thus the construct was capable of specific organic anion transport. Cultured renal epithelial cell lines (MDCK and LLC-PK1) transfected with the vector pEGFP-C3 showed a diffuse, evenly distributed cytoplasmic signal. However, when transfected with pEGFP C3/rROAT1 (vector coding for rROAT1-GFP), both cell lines showed predominantly plasma membrane fluorescence. The expression and distribution of rROAT1-GFP in intact renal proximal tubules was also investigated. Isolated killifish (Fundulus heteroclitus) renal tubules transfected with pEGFP-C3/rROAT1 showed marked basal and lateral membrane-associated fluorescence, but no detectable signal in the nucleus or the apical pole of tubule cells. Tubules transfected with pEGFP-C3 showed diffuse cytoplasmic fluorescence. Function of the rROAT1-GFP construct was demonstrated in transfected killifish tubules by fluorescein transport assay. These results demonstrate the basolateral subcellular localization of rROAT1 in polarized renal epithelia and validate a new technique for localizing cloned transporters within intact renal tubules. PMID- 10362776 TI - Quantification of nitric oxide synthase activity in microdissected segments of the rat kidney. AB - This study was designed to quantify nitric oxide synthase (NOS) activity in microdissected glomeruli (Glm), pars convoluta, pars recta, cortical collecting duct, cortical thick ascending limb, outer medullary collecting duct, medullary thick ascending limb and thin limb, inner medullary collecting duct (IMCD) and thin limb, and vasa recta (VR). Total protein from microdissected segments was incubated with L-[3H]arginine and appropriate cofactors, and the L-arginine and converted L-citrulline were separated by reverse-phase HPLC and radiochemically quantitated. NOS activity was found to be greatest in IMCD (11.5 +/- 1.0 fmol citrulline. mm-1. h-1) and moderate in Glm (1.9 +/- 0.3 fmol. glomerulus-1. h-1) and VR (3.2 +/- 0.8 fmol. mm-1. h-1). All other renal structures studied exhibited significantly less NOS activity. The mRNA for NOS isoforms in the NOS activity-positive segments was then identified by RT-PCR. The IMCD contained mRNA for neuronal (nNOS), endothelial (eNOS), and inducible NOS (iNOS), but Glm and VR only expressed the mRNA for nNOS and eNOS. These experiments demonstrate that the greatest enzymatic activity for NO production in the kidney is in the IMCD, three to sixfold less activity is present in the Glm and VR, and minimal NOS activity is found in other segments studied. PMID- 10362777 TI - Structure and activity of OK-GC: a kidney receptor guanylate cyclase activated by guanylin peptides. AB - Uroguanylin, guanylin, and lymphoguanylin are small peptides that activate renal and intestinal receptor guanylate cyclases (GC). They are structurally similar to bacterial heat-stable enterotoxins (ST) that cause secretory diarrhea. Uroguanylin, guanylin, and ST elicit natriuresis, kaliuresis, and diuresis by direct actions on kidney GC receptors. A 3,762-bp cDNA characterizing a uroguanylin/guanylin/ST receptor was isolated from opossum kidney (OK) cell RNA/cDNA. This kidney cDNA (OK-GC) encodes a mature protein containing 1,049 residues sharing 72.4-75.8% identity with rat, human, and porcine forms of intestinal GC-C receptors. COS or HEK-293 cells expressing OK-GC receptor protein were activated by uroguanylin, guanylin, or ST13 peptides. The 3.8-kb OK-GC mRNA transcript is most abundant in the kidney cortex and intestinal mucosa, with lower mRNA levels observed in urinary bladder, adrenal gland, and myocardium and with no detectable transcripts in skin or stomach mucosa. We propose that OK-GC receptor GC participates in a renal mechanism of action for uroguanylin and/or guanylin in the physiological regulation of urinary sodium, potassium, and water excretion. This renal tubular receptor GC may be a target for circulating uroguanylin in an endocrine link between the intestine and kidney and/or participate in an intrarenal paracrine mechanism for regulation of kidney function via the intracellular second messenger, cGMP. PMID- 10362779 TI - Characterization of Na+/HCO-3 cotransporter isoform NBC-3. AB - Na+-HCO-3 cotransporters mediate the transport of HCO-3 into or out of the cell. Two Na+-HCO-3 cotransporters (NBC) have been identified previously, which are referred to as NBC-1 and NBC-2. A cDNA library from uninduced human NT-2 cells was screened with an NBC-2 cDNA probe. Several clones were identified and isolated. Sequence analysis of these clones identified a partial coding region (2 kb) of a novel NBC (called here NBC-3), which showed 53% and 72% identity with NBC-1 and NBC-2, respectively. Northern blot analysis revealed that NBC-3 encodes a 4.4-kb mRNA with a tissue distribution pattern distinct from NBC-1 and NBC-2. NBC-3 is highly expressed in brain and spinal column, with moderate levels in trachea, thyroid, and kidney. In contrast with NBC-1, NBC-3 shows low levels of expression in pancreas and kidney cortex. In the kidney, NBC-3 expression is predominantly limited to the medulla. Cultured mouse inner medullary collecting duct (mIMCD-3) cells showed high levels of NBC-1 and low levels of NBC-3 mRNA expression. Subjecting the mutagenized mIMCD-3 cells to sublethal acid stress decreased the mRNA expression of NBC-1 by approximately 90% but increased the Na+ dependent HCO-3 cotransport activity by approximately 7-fold (as assayed by DIDS sensitive, Na+-dependent, HCO-3-mediated intracellular pH recovery). This increase was associated with approximately 5.5-fold enhancement of NBC-3 mRNA levels. NBC showed significant affinity for Li+ in the mutant but not the parent mIMCD-3 cells. On the basis of the widespread distribution of NBC-3, we propose that this isoform is likely involved in cell pH regulation by transporting HCO-3 from blood to the cell. We further propose that enhanced expression of NBC-3 in severe acid stress could play an important role in cell survival by mediating the influx of HCO-3 into the cells. PMID- 10362778 TI - Ultrastructural model for size selectivity in glomerular filtration. AB - A theoretical model was developed to relate the size selectivity of the glomerular barrier to the structural characteristics of the individual layers of the capillary wall. Thicknesses and other linear dimensions were evaluated, where possible, from previous electron microscopic studies. The glomerular basement membrane (GBM) was represented as a homogeneous material characterized by a Darcy permeability and by size-dependent hindrance coefficients for diffusion and convection, respectively; those coefficients were estimated from recent data obtained with isolated rat GBM. The filtration slit diaphragm was modeled as a single row of cylindrical fibers of equal radius but nonuniform spacing. The resistances of the remainder of the slit channel, and of the endothelial fenestrae, to macromolecule movement were calculated to be negligible. The slit diaphragm was found to be the most restrictive part of the barrier. Because of that, macromolecule concentrations in the GBM increased, rather than decreased, in the direction of flow. Thus the overall sieving coefficient (ratio of Bowman's space concentration to that in plasma) was predicted to be larger for the intact capillary wall than for a hypothetical structure with no GBM. In other words, because the slit diaphragm and GBM do not act independently, the overall sieving coefficient is not simply the product of those for GBM alone and the slit diaphragm alone. Whereas the calculated sieving coefficients were sensitive to the structural features of the slit diaphragm and to the GBM hindrance coefficients, variations in GBM thickness or filtration slit frequency were predicted to have little effect. The ability of the ultrastructural model to represent fractional clearance data in vivo was at least equal to that of conventional pore models with the same number of adjustable parameters. The main strength of the present approach, however, is that it provides a framework for relating structural findings to the size selectivity of the glomerular barrier. PMID- 10362780 TI - HCO-3 reabsorption in renal collecting duct of NHE-3-deficient mouse: a compensatory response. AB - Mice with a targeted disruption of Na+/H+ exchanger NHE-3 gene show significant reduction in HCO-3 reabsorption in proximal tubule, consistent with the absence of NHE-3. Serum HCO-3, however, is only mildly decreased (P. Schulties, L. L. Clarke, P. Meneton, M. L. Miller, M. Soleimani, L. R. Gawenis, T. M. Riddle, J. J. Duffy, T. Doetschman, T. Wang, G. Giebisch, P. S. Aronson, J. N. Lorenz, and G. E. Shull. Nature Genet. 19: 282-285, 1998), indicating possible adaptive upregulation of HCO-3-absorbing transporters in collecting duct of NHE-3 deficient (NHE-3 -/-) mice. Cortical collecting duct (CCD) and outer medullary collecting duct (OMCD) were perfused, and total CO2 (net HCO-3 flux, JtCO2) was measured in the presence of 10 microM Schering 28080 (SCH, inhibitor of gastric H+-K+-ATPase) or 50 microM diethylestilbestrol (DES, inhibitor of H+-ATPase) in both mutant and wild-type (WT) animals. In CCD, JtCO2 increased in NHE-3 mutant mice (3.42 +/- 0.28 in WT to 5.71 +/- 0.39 pmol. min-1. mm tubule-1 in mutants, P < 0.001). The SCH-sensitive net HCO-3 flux remained unchanged, whereas the DES sensitive HCO-3 flux increased in the CCD of NHE-3 mutant animals. In OMCD, JtCO2 increased in NHE-3 mutant mice (8.8 +/- 0.7 in WT to 14.2 +/- 0.6 pmol. min-1. mm tubule-1 in mutants, P < 0.001). Both the SCH-sensitive and the DES-sensitive HCO 3 fluxes increased in the OMCD of NHE-3 mutant animals. Northern hybridizations demonstrated enhanced expression of the basolateral Cl-/HCO-3 exchanger (AE-1) mRNA in the cortex. The gastric H+-K+-ATPase mRNA showed upregulation in the medulla but not the cortex of NHE-3 mutant mice. Our results indicate that HCO-3 reabsorption is enhanced in CCD and OMCD of NHE-3-deficient mice. In CCD, H+ ATPase, and in the OMCD, both H+-ATPase and gastric H+-K+-ATPase contribute to the enhanced compensatory HCO-3 reabsorption in NHE-3-deficient animals. PMID- 10362781 TI - Serotonin 5-HT2A receptor induces TGF-beta1 expression in mesangial cells via ERK: proliferative and fibrotic signals. AB - We examined the links between fibrotic and proliferative pathways for the 5-HT2A receptor in rat mesangial cells. Serotonin (5-hydroxytryptamine, 5-HT) induced transforming growth factor-beta1 (TGF-beta1) mRNA in a concentration-dependent (peak at 30 nM 5-HT) and time-dependent fashion. For 10 nM 5-HT, the effect was noticeable at 1 h and maximal by 6 h. Inhibition of 1) protein kinase C (PKC), 2) mitogen- and extracellular signal-regulated kinase kinase (MEK1) with 2'-amino-3' methoxyflavone (PD-90859), and 3) extracellular signal-regulated kinase (ERK) with apigenin attenuated this effect. The effect was blocked by antioxidants, N acetyl-L-cysteine (NAC) and alpha-lipoic acid, and mimicked by direct application of H2O2. TGF-beta1 mRNA induction was also blocked by diphenyleneiodonium and 4 (2-aminoethyl)-benzenesulfonyl fluoride, which inhibit NAD(P)H oxidase, a source of oxidants. 5-HT increased the amount of TGF-beta1 protein, validating the mRNA studies and demonstrating that 5-HT potently activates ERK and induces TGF-beta1 mRNA and protein in mesangial cells. Mapping studies strongly supported relative positions of the components of the signaling cascade as follow: 5-HT2A receptor - > PKC --> NAD(P)H oxidase/reactive oxygen species --> MEK --> ERK --> TGF-beta1 mRNA. These studies demonstrate that mitogenic signaling components (PKC, MEK, and oxidants) are directly linked to the regulation of TGF-beta1, a key mediator of fibrosis. Thus a single stimulus can direct both proliferative and fibrotic signals in renal mesangial cells. PMID- 10362782 TI - A numerical model of the renal distal tubule. AB - A numerical model of the rat distal tubule was developed to simulate water and solute transport in this nephron segment. This model incorporates the following: 1) Na-Cl cotransporter, K-Cl cotransporter, Na channel, K channel, and Cl channel in the luminal membrane; 2) Na-K-ATPase, K channel, and Cl channel in the basolateral membrane; and 3) conductances for Na, K, and Cl in the paracellular pathway. Transport rates were calculated using kinetic equations. Axial heterogeneity was represented by partitioning the model into two subsegments with different sets of model parameters. Model equations derived from the principles of mass conservation and electrical neutrality were solved numerically. Values of the model parameters were adjusted to minimize a penalty function that was devised to quantify the difference between model predictions and experimental results. The developed model could simulate the water and solute transport of the distal tubule in the normal state, as well as in conditions including thiazide or amiloride application and various levels of sodium load and tubular flow rate. PMID- 10362783 TI - A kinetic model of the thiazide-sensitive Na-Cl cotransporter. AB - The aim of this study was to construct a numerical model of the thiazide sensitive Na-Cl cotransporter (TSC) that can predict kinetics of thiazide binding and substrate transport of TSC. We hypothesized that the mechanisms underlying these kinetic properties can be approximated by a state diagram in which the transporter has two binding sites, one for sodium and another for chloride and thiazide. On the basis of the state diagram, a system of linear equations that should be satisfied in the steady state was postulated. Numerical solution of these equations yielded model prediction of kinetics of thiazide binding and substrate transport. Rate constants, which determine transitional rates between states, were systematically adjusted to minimize a penalty function that was devised to quantitatively estimate the difference between model predictions and experimental results. With the resultant rate constants, the model could simulate the following experimental results: 1) dissociation constant of thiazide in the absence of sodium and chloride; 2) inhibitory effect of chloride on thiazide binding; 3) stimulatory effect of sodium on thiazide binding; 4) combined effects of sodium and chloride on thiazide binding; 5) dependence of sodium influx on extracellular sodium and chloride; and 6) inhibition of sodium influx by extracellular thiazide. We conclude that known kinetic properties of TSC can be predicted by a model which is based on a state diagram. PMID- 10362784 TI - Genetic instability and the mutator phenotype. Studies in ulcerative colitis. PMID- 10362785 TI - Microglia in Alzheimer's disease and transgenic models. How close the fit? PMID- 10362786 TI - Alport syndrome with diffuse leiomyomatosis. When and when not? PMID- 10362788 TI - Activating c-kit gene mutations in human germ cell tumors. AB - The c-kit gene encodes a tyrosine kinase receptor (KIT) that is required in normal spermatogenesis and is expressed in seminomas and dysgerminomas, a subset of human germ cell tumors (GCTs). To determine whether activating mutations of the c-kit gene occur in GCTs, primary tissue samples of 33 testicular and ovarian tumors were examined for mutations in the juxtamembrane and phosphotransferase domains by polymerase chain reaction amplification and DNA sequencing. A novel missense mutation (D816H) was found in the phosphotransferase domain in tumors of seminoma/dysgerminoma differentiation. The c-kit alleles in nonneoplastic tissues from these patients were wild type, suggesting that the mutant alleles were acquired and selected for during malignant transformation. In cell transfection experiments, the D816H mutant protein was a constitutively activated kinase and was constitutively phosphorylated on tyrosine residues. This is the first description of an activating c-kit mutation in GCTs and is evidence that the KIT signal transduction pathway is important in the pathogenesis of neoplasms with seminoma differentiation. PMID- 10362787 TI - BAT-26 and BAT-40 instability in colorectal adenomas and carcinomas and germline polymorphisms. AB - Analysis of the mononucleotide repeats BAT-26 and BAT-40 has reportedly revealed significant microsatellite instability in sporadic colorectal adenomas, whereas studies with dinucleotide and tetranucleotide repeats have not. In addition, BAT 26 has been reported to be "quasimonomorphic" in the germline. We evaluated BAT 26 and BAT-40 in a series of colorectal tumors previously analyzed with a panel of tetranucleotide repeats. Instability in BAT-26 or BAT-40 was significantly associated with tetranucleotide repeat instability in sporadic adenomas and carcinomas (P < 0.0001) and was similarly much less common in adenomas than in carcinomas. Germline polymorphisms in both BAT-40 and BAT-26 were identified, and the frequency of BAT-26 polymorphisms was significantly higher in African Americans than in Caucasians (7.7% versus 0.08%, P < 0.001). BAT-26 and BAT-40 may be very useful in evaluating instability in small tumors, as sufficient DNA to be amplified by large panels of microsatellites is not always available from such lesions. Polymorphisms in these microsatellites, however, limit their utility in determinations of microsatellite instability without corresponding normal DNA. PMID- 10362789 TI - Stable expression in Chinese hamster ovary cells of mutated tau genes causing frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17). AB - Extensive neuronal loss and aggregation of tau as cytoplasmic inclusions in neurons and glial cells in selected cortical and subcortical regions is the most striking characteristic of frontotemporal dementia and parkinsonism linked to chromosome 17, which is caused by exonic or intronic mutations in the tau gene. Here, we examined the effects of four exonic mutations in four-repeat tau using stably transfected Chinese hamster ovary cells. The proportion of polymerized tubulin was the largest in the P301L transfectant. G272V and P301L transfectants showed greater instability of microtubules in the presence of Colcemid than wild type tau, V337M, or R406W transfectants. Thus no distinct phenotypes were shared by the mutant tau transfectants with regard to microtubule assembly and stability. Unexpectedly, R406W showed low and negligible levels of phosphorylation at Thr 231 and Ser 396, respectively, in the transfectant. This presents a sharp contrast to the observation that tau aggregates in R406W affected brains are heavily phosphorylated at these two sites. This result suggests that hyperphosphorylation at these sites cannot occur in the tau R406W bound to microtubules, and thus that the hyperphosphorylated species of tau may be generated only after disruption of microtubules. PMID- 10362790 TI - Biochemical detection of novel anaplastic lymphoma kinase proteins in tissue sections of anaplastic large cell lymphoma. AB - The (2;5) translocation, found in many T-cell and null cell anaplastic large cell lymphomas (ALCLs), creates a hybrid gene encoding the 80-kd NPM-ALK protein. Typically neoplastic cells show labeling of both nucleus and cytoplasm for anaplastic lymphoma kinase (ALK) and for the N-terminus of nucleophosmin (NPM). However, 10-20% of cases exhibit cytoplasmic labeling only for ALK, indicating the probable presence of variants of the classical (2;5) translocation that do not involve the NPM gene. We report the detection (using Western blotting and an in vitro kinase assay) in seven such ALCL cases, of ALK proteins with molecular masses of 85 kd, 97 kd (one case exhibiting a (2;3)(p23;q21) translocation), 104 kd (one case carried a (1;2)(q21;p23) translocation), and 113 kd. Tyrosine kinase activity was detected in four of these proteins, but the N-terminal portion of NPM could not be detected. These results show how ALCL cases that express ALK proteins other than NPM-ALK can be detected by sensitive biochemical techniques using routine cryostat sections. PMID- 10362791 TI - Frequent loss of KAI1 expression in squamous and lymphoid neoplasms. An immunohistochemical study of archival tissues. AB - The metastasis suppressor gene KAI1 was identified by its ability to inhibit the formation of pulmonary metastases in experimental models for prostatic carcinoma. Down-regulation of this gene may be correlated with the invasive phenotype in melanomas and colon and bladder carcinomas and with the metastatic phenotype in carcinomas of the lung, breast, prostate, and pancreas. The goal of our study was to establish an immunohistochemical method to detect KAI1 expression in archival tissues. Using cell lines with known KAI1 levels and paraffin-embedded KAI1 positive tissues as controls, we observed strong membrane staining in lymphoid follicular centers and squamous epithelia. We then demonstrated the utility of our assay by studying KAI1 expression in 34 lymphoid and 57 squamous lesions. All eight reactive lymph nodes were KAI1 positive. In contrast, three of 13 follicular small cleaved and five of 13 diffuse large cell lymphomas were KAI1 negative. Seventy-nine percent (37 of 47) of invasive squamous cell carcinomas from the lung (n = 15), head and neck (n = 18), and cervix (n = 14) showed extensive KAI1 down-regulation. Loss of KAI1 expression was also found in a subset of 10 high-grade cervical dysplasias. Our data show that (i) immunohistochemistry is a suitable technique for evaluating KAI1 expression in archival tissues; (ii) KAI1 was not expressed in a subset of both low-grade and high-grade lymphomas; and (iii) there was extensive down-regulation of KAI1 in squamous cell carcinomas, suggestive of an important role of the gene in the suppression of invasion in these malignancies. PMID- 10362792 TI - Association of microglia with amyloid plaques in brains of APP23 transgenic mice. AB - Microglia are a key component of the inflammatory response in the brain and are associated with senile plaques in Alzheimer's disease (AD). Although there is evidence that microglial activation is important for the pathogenesis of AD, the role of microglia in cerebral amyloidosis remains obscure. The present study was undertaken to investigate the relationship between beta-amyloid deposition and microglia activation in APP23 transgenic mice which express human mutated amyloid beta precursor protein (betaPP) under the control of a neuron-specific promoter element. Light microscopic analysis revealed that the majority of the amyloid plaques in neocortex and hippocampus of 14- to 18- month-old APP23 mice are congophilic and associated with clusters of hypertrophic microglia with intensely stained Mac-1- and phosphotyrosine-positive processes. No association of such activated microglia was observed with diffuse plaques. In young APP23 mice, early amyloid deposits were already of dense core nature and were associated with a strong microglial response. Ultrastructurally, bundles of amyloid fibrils, sometimes surrounded by an incomplete membrane, were observed within the microglial cytoplasm. However, microglia with the typical characteristics of phagocytosis were associated more frequently with dystrophic neurites than with amyloid fibrils. Although the present observations cannot unequivocally determine whether microglia are causal, contributory, or consequential to cerebral amyloidosis, our results suggest that microglia are involved in cerebral amyloidosis either by participating in the processing of neuron-derived betaPP into amyloid fibrils and/or by ingesting amyloid fibrils via an uncommon phagocytotic mechanism. In any case, our observations demonstrate that neuron derived betaPP is sufficient to induce not only amyloid plaque formation but also amyloid-associated microglial activation similar to that reported in AD. Moreover, our results are consistent with the idea that microglia activation may be important for the amyloid-associated neuron loss previously reported in these mice. PMID- 10362793 TI - High expression of the CC chemokine TARC in Reed-Sternberg cells. A possible explanation for the characteristic T-cell infiltratein Hodgkin's lymphoma. AB - Hodgkin's lymphoma is characterized by the combination of Reed-Sternberg (R-S) cells and a prominent inflammatory cell infiltrate. One of the intriguing questions regarding this disease is what is causing the influx of T lymphocytes into the involved tissues. We applied the serial analysis of gene expression (SAGE) technique on the Hodgkin's lymphoma-derived cell line L428 and on an Epstein-Barr virus (EBV)-transformed lymphoblastoid B-cell line. A frequently expressed tag in L428 corresponded to the T-cell-directed CC chemokine TARC. Reverse transcription polymerase chain reaction analyses demonstrated expression of TARC in nodular sclerosis (NS) and mixed cellularity (MC) classical Hodgkin's lymphomas but not in NLP Hodgkin's lymphoma, anaplastic large-cell lymphomas, and large-B-cell lymphomas with CD30 positivity. Two of five cases of T-cell-rich B cell lymphoma (TCRBCL) were TARC positive. RNA in situ hybridization (ISH) showed a strong signal for TARC in the cytoplasm of R-S cells, and immunohistochemical staining confirmed the presence of the TARC protein in the R-S cells of NS and MC Hodgkin's lymphomas. The lymphocytic and histiocytic (L&H)-type cells of nodular lymphocyte predominance Hodgkin's lymphoma and the neoplastic cells of non Hodgkin's lymphomas with the exception of two cases of TCRBCL did not stain for TARC. TARC is known to bind to the CCR4 receptor, which is expressed on activated Th2 lymphocytes. The immunophenotype of lymphocytes surrounding R-S cells is indeed Th2-like, and by RNA ISH these lymphocytes showed a positive signal for the chemokine receptor CCR4. The findings suggest that production of TARC by the R-S cells may explain the characteristic T-cell infiltrate in classical Hodgkin's lymphoma. PMID- 10362794 TI - Acceleration of c-myc-induced hepatocarcinogenesis by Co-expression of transforming growth factor (TGF)-alpha in transgenic mice is associated with TGF beta1 signaling disruption. AB - We have previously shown in transgenic mice that transforming growth factor (TGF) alpha dramatically enhances c-myc-induced hepatocarcinogenesis by promoting proliferation and survival of hepatocellular carcinoma (HCC) cells. As transgenic livers display increased levels of mature TGF-beta1 from the early stages of hepatocarcinogenesis, we have now assessed whether impairment of TGF-beta1 signaling contributes to the deregulation of cell cycle progression and apoptosis observed during this process. Focal preneoplastic lesions lacking expression of TGF-beta receptor type II (TbetaRII) were detected in c-myc/TGF-alpha but not in c-myc livers. In c-myc/TGF-alpha mice, 40% (2/5) of adenomas and 90% (27/30) of HCCs showed down-regulation of TbetaRII expression in comparison with 11% (2/18) of adenomas and 47% (14/30) of HCCs in c-myc mice. Down-regulation of the TGF beta1-inducible p15(INK4B) mRNA and reduced apoptotic rates in TbetaRII-negative HCCs further indicated the disruption of TGF-beta1 signaling. Furthermore, both TbetaRII-negative and -positive c-myc TGF-alpha HCCs, but not c-myc HCCs, were characterized by decreased levels of the cell cycle inhibitor p27. These results suggest 1) an inverse correlation of decreased p27 expression with the particularly strong expression of TGF-alpha in these lesions, consistent with the capacity of TGF-alpha signaling to post-transcriptionally regulate p27, and 2) the presence of alternative, downstream defects of TGF-beta1 signaling in c myc/TGF-alpha HCCs that may impair the growth-inhibitory response to TGF-beta1. Thus, the accelerated neoplastic development in c-myc/TGF-alpha mice is associated with an early and frequent occurrence of TbetaRII-negative lesions and with reduced levels of p27 in HCC cells, indicating that disruption of TGF-beta1 responsiveness may play a crucial role in the enhancement of c-myc-induced hepatocarcinogenesis by TGF-alpha. PMID- 10362795 TI - Analysis of intracytoplasmic hyaline bodies in a hepatocellular carcinoma. Demonstration of p62 as major constituent. AB - Intracytoplasmic hyaline bodies (IHBs) resemble inclusions in hepatocellular carcinoma cells, which so far have escaped further characterization. A relationship to Mallory bodies was suggested on the basis of light microscopy and filamentous ultrastructure. A hepatocellular carcinoma containing numerous IHBs was studied. Our studies revealed immunoreactivity of IHBs with the monoclonal antibodies SMI 31 and MPM-2, which recognize hyperphosphorylated epitopes present on paired helical filaments in Alzheimer's disease brains (SMI 31) or on diverse proteins hyperphosphorylated by mitotic kinases in the M-phase of the cell cycle (MPM-2). One- and two-dimensional gel electrophoresis of tumor extracts followed by immunoblotting with SMI 31 and MPM-2 antibodies revealed a major immunoreactive protein with an apparent molecular weight between 62 and 65 kd, which was resolved into several highly acidic (pH 4.5) protein components in two dimensional gels. This protein was undetectable in non-neoplastic liver tissue. Sequence analysis identified the SMI 31 and MPM-2 immunoreactive material as p62, indicating that p62 is a major constituent of IHBs. p62 is an only recently discovered protein that is a phosphotyrosine-independent ligand of the SH2 domain of p56(lck), a member of the c-src family of cytoplasmic kinases. Moreover, p62 binds ubiquitin and may act as an adapter linking ubiquitinated species to other proteins. These features suggest a role of p62 in signal transduction and possibly also carcinogenesis. IHBs observed in the hepatocellular carcinoma cells presented are the first indications of a role of p62 in disease. PMID- 10362796 TI - Enhanced expression of CD80 (B7-1), CD86 (B7-2), and CD40 and their ligands CD28 and CD154 in fulminant hepatic failure. AB - To define a possible role for changes in the regulation of antigen presentation in fulminant hepatic failure (FHF), we studied the expression of co-stimulatory molecules CD80 (B7-1), CD86 (B7-2), and CD40 along with their ligands CD28 and CD154. We analyzed the liver tissue from patients with FHF (n = 18), chronic liver disease (n = 30), and acute hepatitis (n = 3) and from normal controls (n = 9) by immunohistochemistry and examined the temporal relationship between CD80/CD86 and CD40 expression and disease in the mouse models of galactosamine lipopolysaccharide and galactosamine-tumor-necrosis-factor-induced FHF. In human controls, faint CD80/CD86 immunoreactivity was restricted to Kupffer cells, and CD40 expression was expressed on bile ducts, macrophages, and sinusoidal endothelial cells (SECs). In FHF, immunoreactivity for CD80 and CD86 was observed on significantly higher numbers of cells, including SECs. Increased CD80/CD86 expression corresponded to increased numbers of CD28-positive lymphocytes. The expression of CD40 was also clearly elevated on virtually all cell types in FHF. In both murine models, CD40 and CD80/CD86 expression was up-regulated before tissue damage could be detected. Our data suggest that up-regulated expression of co-stimulatory molecules might lead to an excessive antigen presentation in FHF as an early step in the pathogenesis before the onset of tissue damage. PMID- 10362797 TI - Coordinate expression of the autosomal dominant polycystic kidney disease proteins, polycystin-2 and polycystin-1, in normal and cystic tissue. AB - A second gene for autosomal dominant polycystic kidney disease (ADPKD), PKD2, has been recently identified. Using antisera raised to the human PKD2 protein, polycystin-2, we describe for the first time its distribution in human fetal tissues, as well as its expression in adult kidney and polycystic PKD2 tissues. Its expression pattern is correlated with that of the PKD1 protein, polycystin-1. In normal kidney, expression of polycystin-2 strikingly parallels that of polycystin-1, with prominent expression by maturing proximal and distal tubules during development, but with a more pronounced distal pattern in adult life. In nonrenal tissues expression of both polycystin molecules is identical and especially notable in the developing epithelial structures of the pancreas, liver, lung, bowel, brain, reproductive organs, placenta, and thymus. Of interest, nonepithelial cell types such as vascular smooth muscle, skeletal muscle, myocardial cells, and neurons also express both proteins. In PKD2 cystic kidney and liver, we find polycystin-2 expression in the majority of cysts, although a significant minority are negative, a pattern mirrored by the PKD1 protein. The continued expression of polycystin-2 in PKD2 cysts is similar to that seen by polycystin-1 in PKD1 cysts, but contrasts with the reported absence of polycystin-2 expression in the renal cysts of Pkd2+/- mice. These results suggest that if a two-hit mechanism is required for cyst formation in PKD2 there is a high rate of somatic missense mutation. The coordinate presence or loss of both polycystin molecules in the same cysts supports previous experimental evidence that heterotypic interactions may stabilize these proteins. PMID- 10362798 TI - Role and localization of urokinase receptor in the formation of new microvascular structures in fibrin matrices. AB - Fibrin or a fibrinous exudate can facilitate angiogenesis in many pathological conditions. In vitro, the outgrowth of capillary-like structures in fibrin can be mimicked by exposing human microvascular endothelial cells (hMVECs) to an angiogenic growth factor and tumor necrosis factor (TNF)-alpha. Urokinase-type plasminogen activator (u-PA) and plasmin activities are required for this angiogenic process. This study focuses on the role and localization of the u-PA receptor (u-PAR) in newly formed microvascular structures. The u-PAR-blocking monoclonal antibody (MAb) H-2 completely inhibited the formation of capillary like tubular structures induced by exposure of hMVECs to basic fibroblast growth factor and TNF-alpha. This was accompanied by a several-fold increase in u-PA accumulation in the conditioned medium. The effect of MAb H-2 was not caused by blocking cellular activation by u-PA/u-PAR interaction, as the amino-terminal fragment (ATF) of u-PA, which also activates u-PAR, prevented tube formation. In addition, the inhibition by MAb H-2 was not due to an effect of the antibody on u PAR-vitronectin binding. These data show that inhibition of tube formation can be caused not only by inhibition of u-PA or plasmin activities but also by unavailability of the u-PAR for cell-bound proteolysis. Immunohistochemical analysis showed that in in vitro angiogenesis u-PAR and u-PA were localized on the invading, tube-forming hMVECs and not on the endothelial cells that are located on top of the fibrin matrix. u-PAR and u-PA were also prominently expressed on endothelial cells of neovessels present in an atherosclerotic plaque. These data may give more insight into the role of u-PAR in repair associated angiogenesis. PMID- 10362799 TI - Dramatic inhibition of retinal and choroidal neovascularization by oral administration of a kinase inhibitor. AB - The most common cause of new blindness in young patients is retinal neovascularization, and in the elderly is choroidal neovascularization. Therefore, there has been a great deal of attention focused on the development of new treatments for these disease processes. Previous studies have demonstrated partial inhibition of retinal neovascularization in animal models using antagonists of vascular endothelial growth factor or other signaling molecules implicated in the angiogenesis cascade. These studies have indicated potential for drug treatment, but have left many questions unanswered. Is it possible to completely inhibit retinal neovascularization using drug treatment with a mode of administration that is feasible to use in patients? Do agents that inhibit retinal neovascularization have any effect on choroidal neovascularization? In this study, we demonstrate complete inhibition of retinal neovascularization in mice with oxygen-induced ischemic retinopathy by oral administration of a partially selective kinase inhibitor that blocks several members of the protein kinase C family, along with vascular endothelial growth factor and platelet derived growth factor receptor tyrosine kinases. The drug also blocks normal vascularization of the retina during development but has no identifiable adverse effects on mature retinal vessels. In addition, the kinase inhibitor causes dramatic inhibition of choroidal neovascularization in a laser-induced murine model. These data provide proof of concept that pharmacological treatment is a viable approach for therapy of both retinal and choroidal neovascularization. PMID- 10362800 TI - Pro-inflammatory cytokines induce expression of matrix-metabolizing enzymes in human cervical smooth muscle cells. AB - The process of cervical ripening has been likened to an inflammatory reaction associated with the catabolism of cervical extracellular matrix by enzymes released from infiltrating leukocytes. We hypothesized that smooth muscle cells in the cervix also participate in this process and that pro-inflammatory cytokines act on cervical smooth muscle cells (CSMC) to provoke the expression of matrix-degrading enzymes. We treated primary cultures of human CSMC with tumor necrosis factor-alpha (TNF-alpha) and examined expression of the elastinolytic enzyme, cathepsin S, the collagen metabolizing matrix metalloproteinases (MMP)-1, -3, -9, and the tissue inhibitor of metalloproteinase (TIMP)-1 and -2. A time course analysis revealed that 10 ng/ml of TNF-alpha induced cathepsin S, MMP-1, 3, and -9 mRNA expression with the maximal response observed after 24-48 hours. TNF-alpha induced cathepsin S, MMP-1, -3, and -9 mRNA expression in a dose dependent manner: the maximal effect was observed at a concentration of 10 ng/ml, with appreciable increases observed at concentrations of 0.1 to 1.0 ng/ml. In contrast, TIMP-1 and -2 mRNAs were not significantly increased by TNF-alpha treatment. Interleukin-1beta produced a pattern of gene expression in the CSMC similar to that observed following TNF-alpha treatment. Western blot analysis and zymography confirmed the induction of proMMP-1, -3, and -9 in response to TNF alpha, but MMP-2 immunoreactivity and zymographic activity were unaffected. TNF alpha increased secretion of procathepsin S, but did not affect TIMP-1 and reduced TIMP-2 production. We conclude that CSMC are targets of pro-inflammatory cytokines, which induce a repertoire of enzymes capable of degrading the cervical extracellular matrix. The induction of these enzymes may facilitate the normal ripening of the cervix at term and participate in the premature cervical changes associated with preterm labor. PMID- 10362801 TI - Emphysematous lesions, inflammation, and fibrosis in the lungs of transgenic mice overexpressing platelet-derived growth factor. AB - Because of its expression pattern and its potent effects on mesenchymal cells, platelet-derived growth factor (PDGF) has been implicated as an important factor in epithelial-mesenchymal cell interactions during normal lung development and in the pathogenesis of fibrotic lung disease. To further explore the role of PDGF in these processes, we have developed transgenic mice that express the PDGF-B gene from the lung-specific surfactant protein C (SPC) promoter. Adult SPC-PDGFB transgenic mice exhibited lung pathology characterized by enlarged airspaces, inflammation, and fibrosis. Emphysematous changes frequently occurred throughout the lung, but inflammation and fibrotic lesions were usually confined to focal areas. The severity of this phenotype varied significantly among individual mice within the same SPC-PDGFB transgenic lineage. A pathology similar to that observed in adult mice was noted in lungs from transgenic mice as young as 1 week of age. Neonatal transgenic mice exhibited enlarged saccules and thickened primary septa. Results of these studies indicated that overexpression of PDGF-B induced distinct abnormalities in the developing and adult lung and led to a complex phenotype that encompassed aspects of both emphysema and fibrotic lung disease. PMID- 10362802 TI - Amplification and overexpression of p40 subunit of eukaryotic translation initiation factor 3 in breast and prostate cancer. AB - Amplification at the long arm of chromosome 8 occurs in a large fraction of breast and prostate cancers. To clone the target genes for this amplification, we used suppression subtraction hybridization to identify overexpressed genes in the breast cancer cell line SK-Br-3, which harbors amplification at 8q (8q21 and 8q23 q24). A differentially expressed gene identified by SSH, the p40 subunit of eukaryotic translation initiation factor 3 (eIF3), was localized to 8q23 and found to be highly amplified and overexpressed in the breast and prostate cancer cell lines studied. High-level amplification of eIF3-p40 was found in 30% of hormone-refractory prostate tumors and in 18% of untreated primary breast tumors. In the vast majority of the cases, p40 and c-myc were amplified with equal copy numbers. Tumors with higher copy numbers of p40 than c-myc were also found. Expression of p40 mRNA was analyzed with in situ hybridization. The amplification of eIF3-p40 gene was associated with overexpression of its mRNA, as expected for a functional target gene of the amplification. These results imply that genomic aberrations of translation initiation factors, such as eIF3-p40, may contribute to the pathogenesis of breast and prostate cancer. PMID- 10362803 TI - Alterations of Fas (Apo-1/CD95) gene in cutaneous malignant melanoma. AB - Fas (Apo-1/CD95) is a cell-surface receptor involved in cell death signaling. The key role of the Fas system in negative growth regulation has been studied mostly within the immune system, and somatic mutations of Fas gene in cancer patients have been described solely in lymphoid-lineage malignancies. However, many nonlymphoid tumor cells have been found to be resistant to Fas-mediated apoptosis, which suggests that Fas mutations, one of the possible mechanisms for Fas resistance, may be involved in the pathogenesis of nonlymphoid malignancies as well. In this study, we have analyzed the entire coding region and all splice sites of the Fas gene for the detection of the gene mutations in 44 human malignant melanomas in skin by polymerase chain reaction, single-strand conformation polymorphism, and DNA sequencing. Overall, 3 tumors (6.8%) were found to have the Fas mutations, which were all missense variants and identified in the cytoplasmic region (death domain) known to be involved in the transduction of an apoptotic signal. The data presented here suggest that somatic alterations of the Fas gene might lead to the loss of its apoptotic function and contribute to the pathogenesis of some human malignant melanomas. PMID- 10362804 TI - Immunohistological analysis of in situ expression of mycobacterial antigens in skin lesions of leprosy patients across the histopathological spectrum. Association of Mycobacterial lipoarabinomannan (LAM) and Mycobacterium leprae phenolic glycolipid-I (PGL-I) with leprosy reactions. AB - The presence of mycobacterial antigens in leprosy skin lesions was studied by immunohistological methods using monoclonal antibodies (MAbs) to Mycobacterium leprae-specific phenolic glycolipid I (PGL-I) and to cross-reactive mycobacterial antigens of 36 kd, 65 kd, and lipoarabinomannan (LAM). The staining patterns with MAb to 36 kd and 65 kd were heterogeneous and were also seen in the lesions of other skin diseases. The in situ staining of PGL-I and LAM was seen only in leprosy. Both antigens were abundantly present in infiltrating macrophages in the lesions of untreated multibacillary (MB) patients, whereas only PGL-I was occasionally seen in scattered macrophages in untreated paucibacillary lesions. During treatment, clearance of PGL-I from granulomas in MB lesions occurred before that of LAM, although the former persisted in scattered macrophages in some treated patients. This persistence of PGL-I in the lesions paralleled high serum anti-PGL-I antibody titers but was not indicative for the presence of viable bacilli in the lesions. Interestingly, we also observed a differential expression pattern of PGL-I and LAM in the lesions of MB patients with reactions during the course of the disease as compared with those without reactions. In conclusion, the in situ expression pattern of PGL-I and LAM in MB patients may assist in early diagnosis of reactions versus relapse. PMID- 10362805 TI - High prevalence of activated intraepithelial cytotoxic T lymphocytes and increased neoplastic cell apoptosis in colorectal carcinomas with microsatellite instability. AB - Microsatellite instability (MSI) characterizes colorectal carcinomas (CRCs) in hereditary nonpolyposis colorectal cancer (HNPCC) syndrome and a proportion of sporadic CRCs. These MSI+ CRCs share several clinicopathological features, including a reputation for better survival rates than MSI- cases and a pronounced stromal inflammatory reaction of still undefined nature. In the present study, the presence, spatial distribution, and activation status of infiltrating cytotoxic effectors were investigated comparatively in 18 MSI+ and 37 MSI- CRCs by immunohistochemistry. The frequency of apoptosis was also evaluated by morphology and in situ end-labeling. MSI+ cases carried significantly higher numbers of cytotoxic lymphocytes infiltrating within neoplastic epithelial structures, as shown by immunostaining for CD3 (15.1 +/- 6.2 versus 4.6 +/- 4.1, P < 0.001), CD8 (13 +/- 6.4 versus 3.7 +/- 3.8, P < 0.001), and TIA-1 (11.2 +/- 6.5 versus 1.9 +/- 1.7, P < 0.001). These cytotoxic effectors were globally more activated in MSI+ than in MSI- tumors, as revealed by the expression of granzyme B (5.3 +/- 4.5 versus 0.6 +/- 1.3, P < 0.001). In MSI+ CRCs, the number of intraepithelial activated cytotoxic lymphocytes was significantly correlated with the proximal location of the tumor, a poorly differentiated phenotype, and the presence of peritumor lymphoid nodules. Multivariate analysis revealed that MSI was the major determinant of the presence of activated cytotoxic intraepithelial lymphocytes. Moreover, MSI+ CRCs also showed a significantly higher percentage of tumor cells undergoing apoptotic cell death (4.1 +/- 2.1 versus 2.6 +/- 1.1, P < 0.0001, by the TUNEL method), often located in close proximity of activated cytotoxic lymphocytes. These results are consistent with the presence of anti tumor cytotoxic immune responses in most of MSI+ CRCs, a phenomenon that may at least in part contribute to the survival advantage ascribed to these patients. PMID- 10362806 TI - Colorectal adenoma and cancer divergence. Evidence of multilineage progression. AB - Colorectal cancer progression involves changes in phenotype and genotype. Although usually illustrated as a linear process, more complex underlying pathways have not been excluded. The object of this paper is to apply modern quantitative principles of molecular evolution to multistep tumor progression. To reconstruct progression lineages, the genotypes of two adjacent adenoma-cancer pairs were determined by serial dilution and polymerase chain reaction at 28-30 microsatellite (MS) loci and then traced back to their most recent common ancestor. The tumors were mismatch repair deficient, and therefore relatively large numbers of MS mutations should accumulate during progression. As expected, the MS genotypes were similar (correlation coefficients >0.9) between different parts of the same adenoma or cancer, but very different (correlation coefficients <0. 2) between unrelated metachronous adenoma-cancer pairs. Unexpectedly, the genotypes of the adjacent adenoma-cancer pairs were also very different (correlation coefficients of 0.30 and 0.36), consistent with early adenoma-cancer divergence rather than direct linear progression. More than 60% of the divisions occurred after this early adenoma-cancer divergence. Therefore, the tumor phylogenies were not consistent with sequential stepwise selection along a single most "fit" and frequent lineage from adenoma to cancer. Instead, one effective early progression strategy creates and maintains multiple evolving candidate lineages, which are subsequently selected for terminal clonal expansion. PMID- 10362807 TI - Genomic instability is an early event during the progression pathway of ulcerative-colitis-related neoplasia. AB - Ulcerative colitis (UC) is a chronic inflammatory disease of the colon associated with a high risk of colorectal cancer. This increased cancer risk is thought to result from the cellular damage induced by the inflammatory field. The aim of this study was to determine the pattern and time course of genomic instability occurring in UC-related neoplasia. Sites of cancer, dysplasia, and nondysplasia from 14 UC colectomy cases containing cancer were analyzed for chromosomal alterations by comparative genomic hybridization (CGH) and for microsatellite instability using a series of 10 microsatellite markers. Clonal chromosomal alterations were present in 85% of cancer sites, 86% of dysplasia sites, and 36% of nondysplasia sites. Losses of chromosome 18 or 18q and chromosome 5 or 5q were common in cancer and dysplasia and were occasionally detected in nondysplasia. High-level microsatellite instability was detected in the cancer and dysplasia of two cases. Samples that demonstrated high-level microsatellite instability were unlikely to have chromosomal alterations demonstrable by CGH. These studies suggest that the predominant type of genomic instability in UC-related neoplasia is associated with chromosomal alterations and that this type of genomic instability frequently occurs before the development of histologically defined dysplasia. PMID- 10362808 TI - Colossal crypts bordering colon adenomas in Apc(Min) mice express full-length Apc. AB - Enlarged but nondysplastic crypts are frequently observed at the margins of colon tumors, forming what has been called a transitional epithelium. It is now thought that this is a reactive state and not a preneoplastic condition as previously suggested. We have used the mouse familial adenomatous polyposis model, ApcMin, to study these abnormal adenoma-associated crypts. We report that these nondysplastic crypts are enormous (as much as 10 times normal length) and branch more frequently than normal crypts. They express wild-type Apc protein and display the wild-type Apc allele. We conclude that the colossal crypts at adenoma margins have normal Apc gene function, consistent with the suggestion that their phenotype is a reactive state. The cause remains an open question, but the dramatic epithelial response hints at the presence of potent epithelial trophic factors in the vicinity of colon tumors. PMID- 10362809 TI - Germline and somatic mutations of the STK11/LKB1 Peutz-Jeghers gene in pancreatic and biliary cancers. AB - Peutz-Jeghers syndrome (PJS) is an autosomal-dominant disorder characterized by hamartomatous polyps in the gastrointestinal tract and by pigmented macules of the lips, buccal mucosa, and digits. Less appreciated is the fact that PJS also predisposes patients to an increased risk of gastrointestinal cancer, and pancreatic cancer has been reported in many PJS patients. It was recently shown that germline mutations of the STK11/LKB1 gene are responsible for PJS. We investigated the role of STK11/LKB1 in the development of pancreatic and biliary cancer in patients with and without the PJS. In a PJS patient having a germline splice site mutation in the STK11/LKB1 gene, sequencing analysis of an intestinal polyp and pancreatic cancer from this patient revealed loss of the wild-type allele of the STK11/LKB1 gene in the cancer. Inactivation of STK11/LKB1, by homozygous deletions or somatic sequence mutations coupled with loss of heterozygosity, was also demonstrated in 4-6% of 127 sporadic pancreatic and biliary adenocarcinomas. Our results demonstrate that germline and somatic genetic alterations of the STK11/LKB1 gene may play a causal role in carcinogenesis and that the same gene contributes to the development of both sporadic and familial forms of cancer. PMID- 10362810 TI - Correlation between clinicopathological features and karyotype in spindle cell sarcomas. A report of 130 cases from the CHAMP study group. AB - Soft-tissue tumors have proved to be a fruitful area for the identification of reproducible cytogenetic aberrations, especially among pediatric round-cell sarcomas and lipomatous tumors. Thus far, however, data regarding sarcomas of monomorphic spindle cell type have been limited and somewhat disappointing, with the notable exception of synovial sarcoma. As part of an ongoing international collaborative study, 130 karyotyped spindle-cell sarcomas were reviewed and classified histologically, without knowledge of the clinical and karyotypic data, with the aim of identifying objective correlations between morphology, karyotype, and clinical parameters. Clonal chromosomal abnormalities were identified in 82 cases studied (63%), but only in the group of synovial sarcomas was there clear correlation between the cytogenetic findings, in the form of a consistent t(X;18)(p11;q11), and morphology. Among leiomyosarcomas (41 cases) and malignant peripheral nerve sheath tumors (MPNSTs; 27 cases) as well as in individual examples of rarer entities, there was a general tendency for karyotypic complexity associated with frequent loss or rearrangement of chromosome arms 1p, 10p, 11q, 12q, 17p, and 22q. Rearrangements of 17q (the region of the NF1 gene) were seen in 9/27 (33%) of MPNSTs. Among nine cases of solitary fibrous tumor (in which previous cytogenetic data are very limited) no consistent aberrations were identified. We conclude that, with the exception of synovial sarcoma, most spindle-cell sarcomas share with pleomorphic sarcomas the tendency for karyotypic complexity. There was no indication (in most of these lesions) that detectable cytogenetic aberrations could either facilitate their diagnosis or help to determine prognosis. There is a clear need to further study and understand the significance of multiple chromosomal abnormalities in this group of mesenchymal neoplasms with the particular goal of determining their role in the process of tumor development. PMID- 10362811 TI - Molecular genetic evidence supporting the clonality and appendiceal origin of Pseudomyxoma peritonei in women. AB - Pseudomyxoma peritonei (PMP) is a poorly understood condition characterized by mucinous ascites and multifocal peritoneal mucinous tumors. Women with PMP often have mucinous tumors involving both the appendix and the ovaries. Several previous histopathological and immunohistochemical studies of PMP have suggested that most, if not all, cases of PMP in women are derived from mucinous adenomas of the appendix rather than from primary ovarian tumors. A few studies of the molecular genetics of PMP have been recently reported. However, these studies analyzed only a small number of cases and some included a heterogeneous group of mucinous tumors, including both benign and malignant appendiceal and ovarian tumors. We analyzed K-ras mutations and allelic losses of chromosomes 18q, 17p, 5q, and 6q in a substantial number of morphologically uniform cases of PMP with synchronous ovarian and appendiceal tumors as well as in appendiceal mucinous adenomas (MAs) and ovarian mucinous tumors of low malignant potential (MLMPs) unassociated with PMP. Each of the 16 PMP cases (100%) analyzed demonstrated identical K-ras mutations in the appendiceal adenoma and corresponding synchronous ovarian tumor. K-ras mutations were identified in 11 of 16 (69%) appendiceal MAs unassociated with PMP and in 12 of 16 (75%) ovarian MLMPs unassociated with PMP. Two PMP cases showed identical allelic losses in the matched ovarian and appendiceal tumors. A discordant pattern of allelic loss between the ovarian and appendiceal tumors at one or two of the loci tested was observed in six PMP cases. In all but one instance, LOH was observed in the ovarian tumor, whereas both alleles were retained in the matched appendiceal lesion, suggesting tumor progression in a secondary (metastatic) site. Our findings strongly support the conclusion that mucinous tumors involving the appendix and ovaries in women with PMP are clonal and derived from a single site, most likely the appendix. PMID- 10362813 TI - Evidence for control of nitric oxide synthesis by intracellular transforming growth factor-beta1 in tumor cells. Implications for tumor development. AB - Transforming growth factor-beta1 (TGF-beta1) has been shown to down-regulate NO synthesis in a variety of normal cells. In the present study, we investigated the influence of TGF-beta1 upon NO production in tumor cells and its consequences for tumor development. During the growth of PROb colon carcinoma cells intraperitoneally injected in syngeneic BDIX rats, intratumoral concentration of TGF-beta1 increases while NO concentration stays very low. Tumor regression induced by intraperitoneal injections of a lipid A is associated with a decrease in TGF-beta1 and an increase in NO intratumoral concentration. In these tumors, PROb tumor cells are the NO- and TGF-beta1-secreting cells. Using PROb cells transfected with an expression vector coding for TGF-beta1 antisense mRNA, we demonstrate in vitro that there is an inverse correlation between the amount of TGF-beta1 secreted and the ability of PROb cells to secrete NO. As the same results were obtained in the presence of an anti-TGF-beta type II receptor neutralizing antibody, and as exogenous TGF-beta1 is without any effect on NO secretion by PROb cells, TGF-beta1 apparently down-regulates NO synthesis in PROb cells by an intracellular mechanism. These results suggest that endogenous TGF beta1 constitutes a potential target in a search for new antitumoral agents. PMID- 10362814 TI - Histological damage in chronic hepatitis C is not related to the extent of infection in the liver. AB - It has not been completely elucidated whether the liver injury induced by the hepatitis C virus (HCV) is due to direct cytopathic damage or to an immune mediated response against HCV-infected hepatocytes. In this work, we have determined the percentage of HCV-infected hepatocytes, the histological activity index, and the viremia levels in chronically HCV-infected patients with different grades of liver injury to investigate any possible correlation between them. For that purpose, liver biopsies from 27 patients with HCV chronic hepatitis were analyzed by in situ hybridization. This technique revealed that the percentage of infected hepatocytes ranged from 0.04% to 83.6%. Regarding the viremia levels, HCV RNA concentration ranged from 1.8 x 10(3) to 1.4 x 10(6) genome copies/ml. A significant correlation (r = 0.54; P = 0.003) between the percentage of infected hepatocytes and the viremia levels was found. In contrast, no correlation was observed between the percentage of HCV-infected hepatocytes or the viremia levels and the histological activity index. In conclusion, we have shown that the HCV viremia reflects the extent of the infection in the liver and that the liver injury in chronic HCV infection is not directly related to either the number of infected hepatocytes or the serum HCV RNA concentration. PMID- 10362812 TI - Composite low grade B-cell lymphomas with two immunophenotypically distinct cell populations are true biclonal lymphomas. A molecular analysis using laser capture microdissection. AB - Low grade B-cell lymphomas comprise several well defined, clinically and immunophenotypically distinct disease entities. Composite lymphomas showing phenotypic characteristics of more than one of these tumor subtypes in the same site are rare, and both common and separate clonal origins of the two tumor parts have been reported for cases studied by molecular methods. We describe the detailed immunohistochemical and molecular findings in three cases with features of composite low grade B-cell non-Hodgkin's lymphoma (B-NHL). All three neoplasms contained morphologically distinct but interwoven compartments of different cell types, which exhibited discordant expression of several markers, including CD5, CD10, CD43, and cyclin D1. According to their morphology and phenotypes, they were classified as mantle cell lymphoma and follicular lymphoma (Case 1), follicular lymphoma and small lymphocytic lymphoma (Case 2), and mantle cell lymphoma and chronic lymphocytic leukemia/small lymphocytic lymphoma (Case 3). PCR analysis of DNA obtained from whole tissue sections failed to reveal evidence for biclonality in any of the cases. We therefore isolated cell populations with different antigen expression patterns by laser capture microdissection and analyzed them by polymerase chain reaction amplification and sequencing of clonal immunoglobulin heavy chain gene rearrangements and oncogene rearrangements. Sequence analysis revealed unrelated clonal rearrangements in each of the two tumor parts in all three cases, suggesting distinct clonal origins. In addition, Case 1 showed a bcl-2 rearrangement present only in the follicular lymphoma part. Our findings suggest that low grade B-NHL with two distinct morphological and immunophenotypic patterns in the same anatomical site are frequently biclonal. This is in keeping with current classification schemes, which recognize subtypes of low grade B-NHL as separate disease entities. Furthermore, our analysis demonstrates the power of laser capture microdissection in revealing molecular microheterogeneity in complex neoplasms. PMID- 10362815 TI - Absence of the alpha6(IV) chain of collagen type IV in Alport syndrome is related to a failure at the protein assembly level and does not result in diffuse leiomyomatosis. AB - X-linked Alport syndrome is a progressive nephropathy associated with mutations in the COL4A5 gene. The kidney usually lacks the alpha3-alpha6 chains of collagen type IV, although each is coded by a separate gene. The molecular basis for this loss remains unclear. In canine X-linked hereditary nephritis, a model for X linked Alport syndrome, a COL4A5 mutation results in reduced mRNA levels for the alpha3, alpha4, and alpha5 chains in the kidney, implying a mechanism coordinating the production of these 3 chains. To examine whether production of alpha6 chain is under the same control, we studied smooth muscle cells from this animal model. We determined the canine COL4A5 and COL4A6 genes are separated by 435 bp, with two first exons for COL4A6 separated by 978 bp. These two regions are >/= 78% identical to the human sequences that have promoter activity. Despite this potential basis for coordinated transcription of the COL4A5 and COL4A6 genes, the alpha6 mRNA level remained normal in affected male dog smooth muscle while the alpha5 mRNA level was markedly reduced. However, both alpha5 and alpha6 chains were absent at the protein level. Our results suggest that production of the alpha6 chain is under a control mechanism separate from that coordinating the alpha3-alpha5 chains and that the lack of the alpha6 chain in Alport syndrome is related to a failure at the protein assembly level, raising the possibility that the alpha5 and alpha6 chains are present in the same network. The lack of the alpha6 chain does not obviously result in disease, in particular leiomyomatosis, as is seen in Alport patients with deletions involving the COL4A5 and COL4A6 genes. PMID- 10362816 TI - McArdle's disease. The unsolved mystery of the reappearing enzyme. AB - We assessed the frequency of muscle fibers showing histochemical phosphorylase activity in 27 muscle biopsies from 25 unrelated patients with McArdle's disease and studied by immunohistochemistry and in situ hybridization whether the muscle specific isoform was expressed. Positive phosphorylase fibers were observed in 19% of our series of biopsies. We show that the enzyme isoform expressed in regenerating fibers differs according to the genotype of patients: the muscle specific isoform is transcribed and translated in patients with none of the described mutations in at least one allele of the myophosphorylase gene, whereas it is neither transcribed nor translated in patients with identified mutations in both alleles. PMID- 10362817 TI - Calcitriol directly sensitizes renal tubular cells to ATP-depletion- and iron mediated attack. AB - Vitamin Ds have been reported to have diverse effects on cell homeostasis, leading to suggestions that they have therapeutic applications extending beyond their traditional actions on the Ca2+/parathyroid/bone axis. As some of these potential indications carry an inherent risk of acute renal failure (ARF; eg, cancer chemotherapy and organ transplantation), the goal of this study was to assess whether vitamin Ds directly affect renal tubule injury responses. Cultured human proximal tubular (HK-2) cells were exposed to physiological or pharmacological doses of either calcitriol (D3) or a synthetic vitamin D2 analogue (19-nor) for 3 to 48 hours. Their impact on cell integrity (percent lactate dehydrogenase (LDH) release and tetrazolium dye MTT uptake) under basal conditions and during superimposed injuries (ATP depletion/Ca2+ ionophore or iron mediated oxidant stress) were determined. As vitamin Ds can be anti proliferative, cell outgrowth ([3H]thymidine uptake and crystal violet staining) was also tested. Finally, the action of D3 on in vivo ARF (glycerol-induced myoglobinuria) and isolated proximal tubule injury responses were assessed. D3 induced a rapid, dose-dependent increase in HK-2 susceptibility to both ATP depletion/Ca2+-ionophore- and Fe-mediated attack without independently affecting cell integrity or proliferative responses. In contrast, D2 negatively affected only Fe toxicity and only after relatively prolonged exposure (48 hours). D3 dramatically potentiated in vivo ARF (two- to threefold increase in azotemia), suggesting potential in vivo relevance of the above HK-2 cell results. Proximal tubules, isolated from these glycerol-exposed mice, suggested that D3 can worsen tubule injury despite a parodoxic suppression of H2O2 production. In contrast, D3 had a mild negative impact on cellular energetics (depressed ATP/ADP ratios), and it accentuated plasma membrane phospholipid breakdown. The latter was observed in both glycerol-treated and control tubules, suggesting a primary role in the injury- potentiation effect of D3. Vitamins D(s) may directly, and differentially, increase proximal tubule cell susceptibility to superimposed attack. This property should be considered as new uses for these agents are defined. PMID- 10362818 TI - Shifts in lung lymphocyte profiles correlate with the sequential development of acute allergic and chronic tolerant stages in a murine asthma model. AB - T lymphocytes have a central regulatory role in the pathogenesis of asthma. We delineated the participation of lymphocytes in the acute allergic and chronic tolerant stages of a murine model of asthma by characterizing the various subsets of lymphocytes in bronchoalveolar lavage and lung tissue associated with these responses. Acute (10-day) aerosol challenge of immunized C57BL/6J mice with ovalbumin resulted in airway eosinophilia, histological evidence of peribronchial and perivascular airway inflammation, clusters of B cells and TCRgammadelta cells in lung tissue, increased serum IgE levels, and airway hyperresponsiveness to methacholine. In mice subjected to chronic (6-week) aerosol challenge with ovalbumin, airway inflammation and serum IgE levels were significantly attenuated and airway hyperresponsiveness was absent. The marked increases in lung B and T cell populations seen in the acute stage were also significantly reduced in the chronic stage of this model. Thus, acute ovalbumin challenge resulted in airway sensitization characteristic of asthma, whereas chronic ovalbumin challenge elicited a suppressed or tolerant state. The transition from antigenic sensitization to tolerance was accompanied by shifts in lymphocyte profiles in the lung and bronchoalveolar lavage fluid. PMID- 10362819 TI - A nonhuman primate model for the selective elimination of CD8+ lymphocytes using a mouse-human chimeric monoclonal antibody. AB - Nonhuman primates provide valuable animal models for human diseases. However, studies assessing the role of cell-mediated immune responses have been difficult to perform in nonhuman primates. We have shown that CD8+ lymphocyte-mediated immunity in rhesus monkeys can be selectively eliminated using the mouse-human chimeric anti-CD8 monoclonal antibody cM-T807. In vitro, this antibody completely blocked antigen-specific expansion of cytotoxic T cells and decreased major histocompatibility complex class I-restricted, antigen-specific lysis of target cells but did not mediate complement-dependent cell lysis. In vivo administration of cM-T807 in rhesus monkeys resulted in near total depletion of CD8+ T cells from the blood and lymph nodes for up to 6 weeks. This depletion was not solely complement-dependent and persisted longer in adults than in juveniles. Preservation of B cell and CD4+ T cell function in monkeys depleted of CD8+ lymphocytes was demonstrated by their ability to develop humoral immune responses to the administered chimeric monoclonal antibody. Furthermore, during CD8+ lymphocyte depletion, monkeys developed delayed-type hypersensitivity reactions comprised only of CD4+ T cells but not CD8+ T cells. This CD8+ lymphocyte depletion model should prove useful in defining the role of cell-mediated immune responses in controlling infectious diseases in nonhuman primates. PMID- 10362820 TI - Effect of cigar smoking on the risk of cardiovascular disease, chronic obstructive pulmonary disease, and cancer in men. AB - BACKGROUND: The sale of cigars in the United States has been increasing for the past six years. Cigar smoking is a known risk factor for certain cancers and for chronic obstructive pulmonary disease (COPD). However, unlike the relation between cigarette smoking and cardiovascular disease, the association between cigar smoking and cardiovascular disease has not been clearly established. METHODS: We performed a cohort study among 17,774 men 30 to 85 years of age at base line (from 1964 through 1973) who were enrolled in the Kaiser Permanente health plan and who reported that they had never smoked cigarettes and did not currently smoke a pipe. Those who smoked cigars (1546 men) and those who did not (16,228) were followed from 1971 through the end of 1995 for a first hospitalization for or death from a major cardiovascular disease or COPD, and through the end of 1996 for a diagnosis of cancer. RESULTS: In multivariate analysis, cigar smokers, as compared with nonsmokers, were at higher risk for coronary heart disease (relative risk, 1.27; 95 percent confidence interval, 1.12 to 1.45), COPD (relative risk, 1.45; 95 percent confidence interval, 1.10 to 1.91), and cancers of the upper aerodigestive tract (relative risk, 2.02; 95 percent confidence interval, 1.01 to 4.06) and lung (relative risk, 2.14; 95 percent confidence interval, 1.12 to 4.11), with evidence of dose-response effects. There appeared to be a synergistic relation between cigar smoking and alcohol consumption with respect to the risk of oropharyngeal cancers and cancers of the upper aerodigestive tract. CONCLUSIONS: Independently of other risk factors, regular cigar smoking can increase the risk of coronary heart disease, COPD, and cancers of the upper aerodigestive tract and lung. PMID- 10362821 TI - Effects of tissue plasminogen activator for acute ischemic stroke at one year. National Institute of Neurological Disorders and Stroke Recombinant Tissue Plasminogen Activator Stroke Study Group. AB - BACKGROUND: In 1995, the two-part National Institute of Neurological Disorders and Stroke (NINDS) Recombinant Tissue Plasminogen Activator Stroke Trial found that patients who were treated with tissue plasminogen activator (t-PA) within three hours after the onset of symptoms of acute ischemic stroke were at least 30 percent more likely than patients given placebo to have minimal or no disability three months after the stroke. It was unknown, however, whether the benefit would be sustained for longer periods. METHODS: In the NINDS Trial, a total of 624 patients with stroke were randomly assigned to receive either t-PA or placebo. We collected outcome data over a period of 12 months after the occurrence of stroke. The primary outcome measure was a "favorable outcome," defined as minimal or no disability as measured by the Barthel index, the modified Rankin Scale, and the Glasgow Outcome Scale. We assessed the treatment effect using a global statistic. RESULTS: Using an intention-to-treat analysis for the combined results of the two parts of the trial at 6 months and 12 months, we found that the global statistic favored the t-PA group (odds ratio for a favorable outcome at 6 months, 1.7; 95 percent confidence interval, 1.3 to 2.3; odds ratio at 12 months, 95 percent confidence interval, 1.7; 1.2 to 2.3). The patients treated with t-PA were at least 30 percent more likely to have minimal or no disability at 12 months than were the placebo-treated patients (absolute increase in the proportion with a favorable outcome, 11 to 13 percentage points). There was no significant difference in mortality at 12 months between the t-PA group and the placebo group (24 percent vs. 28 percent, P=0.29). There was no interaction between the type of stroke identified at base line and treatment with respect to the long-term response. The rate of recurrent stroke at 12 months was similar in the two groups. CONCLUSIONS: During 12 months of follow-up, the patients with acute ischemic stroke who were treated with t-PA within three hours after the onset of symptoms were more likely to have minimal or no disability, than the patients given placebo. These results indicate a sustained benefit of t-PA for such patients. PMID- 10362823 TI - Time of implantation of the conceptus and loss of pregnancy. AB - BACKGROUND: Implantation of the conceptus is a key step in pregnancy, but little is known about the time of implantation or the relation between the time of implantation and the outcome of pregnancy. METHODS: We collected daily urine samples for up to six months from 221 women attempting to conceive after ceasing to use contraception. Ovulation was identified on the basis of the ratio of urinary estrogen metabolites to progesterone metabolites, which changes rapidly with luteinization of the ovarian follicle. The time of implantation was defined by the appearance of chorionic gonadotropin in maternal urine. RESULTS: There were 199 conceptions, for 95 percent of which (189) we had sufficient data for analysis. Of these 189 pregnancies, 141 (75 percent) lasted at least six weeks past the last menstrual period, and the remaining 48 pregnancies (25 percent) ended in early loss. Among the pregnancies that lasted six weeks or more, the first appearance of chorionic gonadotropin occurred 6 to 12 days after ovulation; 118 women (84 percent) had implantation on day 8, 9, or 10. The risk of early pregnancy loss increased with later implantation (P<0.001). Among the 102 conceptuses that implanted by the ninth day, 13 percent ended in early loss. This proportion rose to 26 percent with implantation on day 10, to 52 percent on day 11, and to 82 percent after day 11. CONCLUSIONS: In most successful human pregnancies, the conceptus implants 8 to 10 days after ovulation. The risk of early pregnancy loss increases with later implantation. PMID- 10362822 TI - A serologic marker of paraneoplastic limbic and brain-stem encephalitis in patients with testicular cancer. AB - BACKGROUND: In patients with cancer, symptoms of limbic and brain-stem dysfunction may result from a paraneoplastic disorder. Paraneoplastic limbic or brain-stem encephalitis occurs more frequently with testicular cancer than with most other cancers. We sought antineuronal antibodies that might be used in a diagnostic test for this syndrome. METHODS: Immunohistochemical and immunoblotting techniques were used to detect serum and cerebrospinal fluid antibodies. Serologic screening of a complementary DNA library and Northern blotting were used to clone the target antigen and determine which tissues expressed it. RESULTS: Of 13 patients with testicular cancer and paraneoplastic limbic or brain-stem encephalitis (or both), 10 had antibodies in serum and cerebrospinal fluid against a 40-kd neuronal protein. These antibodies were used to clone a gene that we call Ma2, which codes for a protein (Ma2) that was recognized by serum from the 10 patients, but not by serum from 344 control subjects. Ma2 was selectively expressed by normal brain tissue and by the testicular tumors of the patients. Ma2 shares homology with Ma1, a "brain-testis cancer" gene related to other paraneoplastic syndromes and tumors. CONCLUSIONS: The serum of patients with subacute limbic and brain-stem dysfunction and testicular cancer contains antibodies against a protein found in normal brain and in testicular tumors. Detection of these antibodies supports the paraneoplastic origin of the neurologic disorder and could be of diagnostic importance. PMID- 10362824 TI - Images in clinical medicine. Hereditary hemorrhagic telangiectasia (Osler-Weber Rendu disease). PMID- 10362825 TI - The protective effects of estrogen on the cardiovascular system. PMID- 10362826 TI - Arteriovenous malformations of the brain in adults. PMID- 10362827 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 17-1999. A 42-year-old asplenic man with gram-negative sepsis. PMID- 10362828 TI - The NIH "E-biomed" proposal--a potential threat to the evaluation and orderly dissemination of new clinical studies. PMID- 10362829 TI - Cigars and public health. PMID- 10362830 TI - The importance of defining the paraneoplastic neurologic disorders. PMID- 10362831 TI - The toll of innate immunity on microbial pathogens. PMID- 10362832 TI - Analysis of the expression and enzymatic properties of alpha1- >3fucosyltransferase from human lung carcinoma NCI-H69 and PC9 cells. AB - An analysis of alpha1-->3fucosyltransferase expression and enzyme properties has been conducted in human lung carcinoma NCI-H69 and PC9 cells. The results indicate that multiple forms of alpha1-->3 fucosyltransferase are found in these cells. RT-PCR experiments using total RNA from NCI-H69 and PC9 cells amplified transcripts for three of these enzymes, FucT-IV, -VI, and -VII. Fucose transfer into glycolipid acceptors mediated by truncated chimeric and full length recombinant FucT-IV and -VI enzymes was examined. Both enzymes were found to be type 2 chain specific, but only FucT-VI efficiently transferred fucose to both neutral and sialylated acceptors. A truncated recombinant form of FucT-VI was capable of fucose transfer to the internal Glc residue of a variety of glycolipid acceptors. This property was not observed with the recombinant full length enzyme, suggesting the N-terminal portion of the protein, composed of the intracellular domain, transmembrane domain, and a part of the stem region, is involved in interactions with glycolipid acceptors. Using taurodeoxycholate as the detergent, the distribution of initial fucose transfer into nLc6catalyzed by recombinant full length enzyme indicated 34% of the mono-fucosyl product was fucosylated at the III-GlcNAc and 66% at the V-GlcNAc for FucT-IV, and almost all of the FucT-VI mono-fucosyl product was III-GlcNAc fucosylated. Similar experiments with VI2NeuAcnLc6as the acceptor resulted in predominantly III-GlcNAc monofucosylation, although detectable V-GlcNAc monofucosylation was obtained with FucT-VI. When the cationic detergent G-3634-A was used, substantially greater initial transfer into the V-GlcNAc of both neutral and sialylated acceptors with FucT-VI was observed. Using nonsialylated acceptors, total alpha1-->3 fucosyltransferase activity in NCI-H69 cells was analyzed and found to be diminished 25-30% by exposure to 30 mM NEM, which can be attributed to FucT-VI inactivation. The remaining 70-75% of NEM-resistant activity is attributed to FucT-IV, an NEM-resistant enzyme form capable of fucosylating nonsialylated acceptors. These results suggest that multiple forms of alpha1- >3fucosyltransferase are expressed in NCI-H69 and PC9 cells, which may account for the observed properties of enzyme derived from these cell lines. PMID- 10362833 TI - Use of inhibitors to characterize intermediates in the processing of N-glycans synthesized by insect cells: a metabolic study with Sf9 cell line. AB - The most frequent type of N-glycan synthesized by lepidopteran Sf9 cells appears to be fucosylated Man3GlcNAc2,and this has been a limitation for a large scale production and utilization of therapeutic glycoproteins in cultured insect cells. The current knowledge of the protein glycosylation pathway derived from structural studies on recombinant glyco-proteins expressed by using baculovirus vectors. In this work we provide more direct evidence for the sequential events occurring in the processing of endogenous N-glycoproteins of noninfected Sf9 cells. By metabolic labeling with radioactive mannose, we characterized the glycan structures which accumulated in the presence of processing inhibitors (castanospermine and swainsonine) and in the presence of an intracellular trafficking inhibitor (monensin). We thus demonstrated that from the glycan precursor Glc3Man9GlcNAc2 to GlcNAcMan5(Fuc)GlcNAc2 intermediate, the processing pathway in Sf9 cells paralleled the one demonstrated in mammalian cells. By using monensin, we demonstrated the formation of Man3(Fuc)GlcNAc2 from GlcNAcMan3(Fuc)GlcNAc2, a reaction which has not been described in mammalian cells. Our results support the idea that the hexosaminidase activity is of physiological relevance to the glycosylation pathway and is Golgi located. PMID- 10362834 TI - Evidence for the presence of major peripheral myelin glycoprotein P0 in mammalian spinal cord and a change of its glycosylation state during aging. AB - Glycoproteins, which react with Lens culinaris agglutinin, in the membrane preparation of various portions of brains and spinal cords, obtained from 9-week old rats and 29-month-old rats, were comparatively analyzed by SDS-polyacrylamide gel electrophoresis. In contrast to the samples from brain, which showed similar staining patterns in the two different age groups, the glycoprotein patterns of spinal cords showed marked differences by the age of donors. The most prominent evidence is that a glycoprotein with an apparent molecular weight of 30 kDa (gp30) was detected in the aged rats, but not in the young adult rats. Based on the amino acid sequence data around the glycosylation site, the gp30 was identified as P0, which is a member of immunoglobulin superfamily and a major structural component of mammalian peripheral nerve myelin. This is the first report indicating that P0, which has been considered as a peripheral nerve specific glycoprotein, occurs also in the spinal cord of mammals. In addition, nonglycosylated P0 molecule could be detected in the spinal cord of young adult rats by anti-P0 polyclonal antibody. These results indicate that the glycosylation state of the P0 molecule in the spinal cord changes during aging. PMID- 10362835 TI - Characterization of the substrate specificity of alpha1,3galactosyltransferase utilizing modified N-acetyllactosamine disaccharides. AB - alpha1,3galactosyltransferase (alpha1,3GalT) catalyzes the synthesis of a range of glycoconjugates containing the Galalpha1,3Gal epitope which is recognized by the naturally occurring human antibody, anti-Gal. This enzyme may be a useful synthetic tool to produce a range of compounds to further investigate the binding site of anti-Gal and other proteins with a Galalpha1,3Gal binding site. Thus, the enzyme has been probed with a series of type 2 disaccharide-C8(Galbeta1-4GlcNAc C8) analogs. The enzyme tolerated acceptors with modifications at C2 and C3 of the N-acetylglucosamine residue, producing a family of compounds with a nonreducing alpha1,3 linked galactose. Compounds that did not serve as acceptors were evaluated as inhibitors. Interestingly, the type 1 disaccharide-C8, Galbeta1 3GlcNAc-C8, was a good inhibitor of the enzyme (Ki = 270 microM vs. Km = 190 microM for Galbeta1-4GlcNAc-C8). A potential photoprobe, based on a modified type 2 disaccharide (octyl 3-amino-3-deoxy-3-N-(2-diazo-3, 3, 3-trifluoropropionyl beta-D-galactopyranosyl-(1, 4)-2-acetamindo-2-deoxy-beta-D-glycopyranoside, (DTFP LacNAc-C8)), was evaluated as an inhibitor of alpha1,3GalT. alpha1,3GalT bound DTFP-LacNAc-C8 with an affinity (Ki = 300 microM) similar to that displayed by the enzyme for LacNAc-C8. Additional studies were done to determine the enzyme's ability to transfer a range of sugars from UDP-sugar donors. The results of these experiments demonstrated that alpha1,3GalT has a strict specificity for UDP-Gal. Finally, inactivation studies with various amino acid modifiers were done to obtain information on the importance of different types of amino acids for alpha1,3GalT activity. PMID- 10362836 TI - NMR and molecular modeling studies on two glycopeptides from the carbohydrate protein linkage region of connective tissue proteoglycans. AB - Complete 1H and 13C NMR assignments are reported for two glycopeptides representing the carbohydrate-protein linkage region of connective tissue proteoglycans. These glycopeptides are the octasaccharide hexapeptide, Ser(GlcpAbeta(1-->3) Galpbeta(1-->3)Galpbeta(1-->4)Xylpbeta)-Gly-Ser-Gly-Se r (GlcpAbeta(1-->3)Galpbeta(1-->3)Galpbeta(1-->4)Xylp beta)-Gly (1), and the tetrasaccharide dipeptide, Ser(GlcpAbeta(1-->3)Galpbeta(1-->3)Galpbeta(1-->4)X ylpbeta)-Gly (2). The vicinal coupling constant data show that the monosaccharide residues adopt4 C 1 chair conformations. Distance geometry/simulated annealing calculations using 2D NOESY derived distance constraints yielded a single family of structures for the tetrasaccharide moiety, with well defined interglycosidic linkage conformations. The straight phi torsion angles of the glycosidic C1'-O1 bonds showed a strict preference for the -sc range whereas the psi torsion angles (O1-Cn) exhibited dependence upon the interglycosidic linkage position (-ac for beta(1-->3) linkage, +ac for beta(1-->4) linkage). The predominant conformation about the glycopeptide bond is straight phi = -sc and psi = +ac. The presence of strong daN (i, i+1) NOE contacts, and the general absence of dNN (i, i+1) contacts (except for a weak Ser-5/Gly-6 dNN contact) and the dbN (i, i+1) contacts (except for Ser-1/Gly-2) in the ROESY spectrum, suggest that the backbone for 1 is predominantly in an extended conformation. A comparison of the ROESY data for 1 with those obtained from the unglycosylated hexapeptide (3) of the same sequence suggests that glycosylation has only a marginal influence on the backbone conformation of the hexapeptide. PMID- 10362837 TI - The O-linked fucose glycosylation pathway: identification and characterization of a uridine diphosphoglucose: fucose-beta1,3-glucosyltransferase activity from Chinese hamster ovary cells. AB - O-Linked fucose is an unusual carbohydrate modification in which fucose is linked directly to the hydroxyl groups of serines or threonines. It has been found on the epidermal growth factor-like modules of several secreted proteins involved in blood coagulation and fibrinolysis. We have recently reported the existence of an elongated form of O-linked fucose in Chinese hamster ovary cells consisting of a glucose linked to the 3'-hydroxyl of fucose (Glcbeta1,3Fuc- O-Ser/Thr). This structure is highly unusual for two reasons. First, in mammalian systems fucose is usually a terminal modification of N - and O-linked oligosaccharides. Here the fucose is internal. Secondly, terminal beta-linked glucose is extremely rare on mammalian glycoconjugates. Thus, the Glcbeta1,3Fuc structure is a very unique mammalian carbohydrate structure. Here we report the identification and initial characterization of a novel enzyme activity capable of forming this unique linkage: UDP-glucose: O-linked fucose beta1,3 glucosyltransferase. The enzyme utilizes UDP-glucose as the high energy donor and transfers glucose to alpha linked fucose residues. The activity is linearly dependent on time, enzyme, and substrate concentrations and is enhanced in the presence of manganese ions. Activity is present in extracts of cultured cells from a variety of species (hamster, human, mouse, rat, chicken) and is enriched in brain and spleen of a normal adult rat. Thus, while this glycosyltransferase appears to be widespread in biology, it forms a very unique linkage, and it represents the first mammalian enzyme identified capable of elongating fucose. PMID- 10362838 TI - Reevaluating the effect of Brefeldin A (BFA) on ganglioside synthesis: the location of GM2 synthase cannot be deduced from the inhibition of GM2 synthesis by BFA. AB - Brefeldin A reversibly disassembles the Golgi complex, causing mixing of the Golgi cisternae with the ER while the trans Golgi network persists as part of a separate endosomal membrane system. Because of this compartmental separation, Brefeldin A treatment has been used to map the sub-Golgi locations of several Golgi enzymes including GM2 synthase. We previously proposed that GM2 synthase might be located in a distal portion of the Golgi complex which in the presence of Brefeldin A would be separated from the substrate ganglioside GM3 present in the mixed ER-Golgi membrane system. In the present study we show using GM2 synthase chimeras that GM2 synthesis was blocked by Brefeldin A when GM2 synthase was distributed throughout all Golgi subcompartments or even when it was restricted to the medial Golgi. Because these findings opposed our speculation regarding a distal location of this enzyme, we sought an alternative explanation for the inhibition of ganglioside synthesis by Brefeldin A. However, Brefeldin A did not degrade GM2 synthase, prevent its homodimerization, or inhibit its in vitro activity. Brefeldin A did result in the conversion of a portion of membrane bound GM2 synthase into a soluble form which has minimal capability to produce GM2 in whole cells. However, this conversion was not sufficient to explain the nearly total loss of GM2 production in intact cells in the presence of Brefeldin A. Nevertheless, the results of this study indicate that Brefeldin A-induced inhibition of ganglioside synthesis cannot be used to deduce the location of GM2 synthase. PMID- 10362839 TI - Identification and characterization of a novel UDP-GalNAc:GlcAbeta-R alpha1,4-N acetylgalactosaminyltransferase from a human sarcoma cell line. AB - We recently discovered a novel alpha-N-acetylgalactosaminyltransferase in fetal bovine serum (Kitagawa et al., J. Biol. Chem., 270, 22190-22195, 1995) and also in mouse mast cytoma cells (Lidholt et al., Glycoconjugate J., 14, 737-742, 1997), which catalyzed the transfer of an alpha-GalNAc residue to the linkage tetrasaccharide-serine, GlcAbeta1-3Galbeta1-3Galbeta1-4Xylbeta1-O-Ser, derived from proteoglycans. In this study, we characterized this enzyme using a preparation obtained from the serum-free culture medium of a human sarcoma (malignant fibrous histiocytoma) cell line by phenyl-Sepharose chromatography. Structural characterization by1H NMR spectroscopy of the reaction product using the linkage tetrasaccharide-serine, GlcAbeta1-3Galbeta1-3Galbeta1-4Xylbeta1-O Ser, as a substrate demonstrated that the enzyme was a UDP-GalNAc:GlcAbeta1-R alpha1,4-N -acetylgalactosaminyltransferase. This is the first identification of an alpha1,4-N-acetylgalactosaminyltransferase. Using N -acetylchondrosine GlcAbeta1-3GalNAc as an alternative substrate, the enzyme required divalent cations for the transferase reaction, with maximal activity at 20 mM Mn2+and exhibited a dual optimum at pH 6.5 and pH 7.4 depending upon the buffers used, with the highest activity in a 50 mM 2-( N -morpholino)ethanesulfonic acid buffer at pH 6.5. The apparent Km values obtained for N -acetylchondrosine, the linkage tetrasaccharide-serine, and UDP-GalNAc were 1060 microM, 188 microM, and 27 microM, respectively. This suggested that the linkage tetrasaccharide-serine was a good acceptor substrate for the enzyme. In addition, the enzyme utilized glucuronylneolactotetraosylceramide GlcAbeta1-3Galbeta1-4GlcNAcbeta1-3Galbeta1-4G lcbeta1-1Cer but not sulfoglucuronylneolactotetraosylceramide GlcA(3-O sulfate)beta1-3Galbeta1-4GlcNAcbeta1-3Galbeta1-4Gl cbeta1-1Cer as acceptor substrates. The possibility of involvement of this enzyme in the biosynthesis of glycosaminoglycan as well as other GlcA-containing glycoconjugates is discussed. PMID- 10362840 TI - Structural features in heparin that interact with VEGF165 and modulate its biological activity. AB - The 165 amino acid form of vascular endothelial growth factor (VEGF165) is a heparin-binding growth factor with mitogenic activity for vascular endothelial cells. We examined activities of various heparin derivatives toward their interactions with VEGF165 using an enzyme-linked immunosorbent assay and elucidated the structural features in heparin for the interactions. Native heparin interacted with VEGF165, whereas N-desulfated, N-acetylated (N-DS, N-Ac-) heparin, and 6-O-desulfated (6-O-DS-) heparin did not. The 2-O-desulfated (2-O-DS ) heparin retained the ability for the interaction with VEGF165. In contrast, the 2-O-DS-heparin exhibited no ability for the interaction with FGF-2 and HGF. Thus, structural requirements in heparin for the specific interaction with VEGF165 are distinct from those with FGF-2 and HGF which require a high content of 2-O sulfate groups. In a cell proliferation assay, native heparin and 2-O-DS-heparin exhibited inhibitory abilities for VEGF165-induced proliferation of human umbilical vein endothelial cells (HUVECs) with their high concentrations (more than 64 microg/ml), while only native heparin could enhance the proliferation of the chlorate-treated cells. These results suggested that a high content of 2-O sulfate groups is not required for the specific interaction with VEGF165alone, although it is essential for the mitogenic activity of the growth factor. PMID- 10362841 TI - Fold recognition study of alpha3-galactosyltransferase and molecular modeling of the nucleotide sugar-binding domain. AB - The structure and fold of the enzyme responsible for the biosynthesis of the xenotransplantation antigen, namely pig alpha3 galactosyltransferase, has been studied by means of computational methods. Secondary structure predictions indicated that alpha3-galactosyltransferase and related protein family members, including blood group A and B transferases and Forssman synthase, are likely to consist of alternating alpha-helices and beta-strands. Fold recognition studies predicted that alpha3-galactosyltransferase shares the same fold as the T4 phage DNA-modifying enzyme beta-glucosyltransferase. This latter enzyme displays a strong structural resemblance with the core of glycogen phosphorylase b. By using the three-dimensional structure of beta-glucosyltransferase and of several glycogen phosphorylases, the nucleotide binding domain of pig alpha3 galactosyltransferase was built by knowledge-based methods. Both the UDP galactose ligand and a divalent cation were included in the model during the refinement procedure. The final three-dimensional model is in agreement with our present knowledge of the biochemistry and mechanism of alpha3 galactosyltransferases. PMID- 10362842 TI - Characterization of fibroblast growth factor 1 binding heparan sulfate domain. AB - Fibroblast growth factors FGF-1 and FGF-2 mediate their biological effects via heparan sulfate-dependent interactions with cell surface FGF receptors. While the specific heparan sulfate domain binding to FGF-2 has been elucidated in some detail, limited information has been available concerning heparan sulfate structures involved in the recognition of FGF-1. In the current study we present evidence that the minimal FGF-1 binding heparan sulfate sequence comprises 5-7 monosaccharide units and contains a critical trisulfated IdoA(2-OSO3)-GlcNSO3(6 OSO3) disaccharide unit. N-Sulfated heparan sulfate decasaccharides depleted of FGF-1 binding domains showed dose-dependent and saturable binding to FGF-2. These data indicate that the FGF-1 binding domain is distinct from the minimal FGF-2 binding site, previously shown to contain an IdoA(2-OSO3) residue but no 6-O sulfate groups. We further show that the FGF-1 binding heparan sulfate domain is expressed in human aorta heparan sulfate in an age-related manner in contrast to the constitutively expressed FGF-2 binding domain. Reduction of heparan sulfate O sulfation by chlorate treatment of cells selectively impedes binding to FGF-1. The present data implicate the 6-O-sulfation of IdoA(2-OSO3)-GlcNSO3 units in cellular heparan sulfate in the control of the biological activity of FGF-1. PMID- 10362843 TI - O-Glycosylation of the V2 vasopressin receptor. AB - The human V2 vasopressin receptor contains one consensus site for N-linked glycosylation at asparagine 22 in the predicted extracellular amino terminal segment of the protein. This segment also contains clusters of serines and threonines that are potential sites for O-glycosylation. Mutagenesis of asparagine 22 to glutamine abolished N-linked glycosylation of the V2 receptor (N22Q-V2R), without altering its function or level of expression. The N22Q-V2R expressed in transfected cells migrated in denaturing acrylamide gels as two protein bands with a difference of 7000 Da. Protein labeling experiments demonstrated that the faster band could be chase to the slower one suggesting the presence of O-linked sugars. Sialidase treatment of membranes from cells expressing the N22Q-V2R or of immunoprecipitated metabolically labeled V2R accelerated the migration of the protein in acrylamide gels demonstrating the existence of O-glycosylation, the first time this type of glycosylation has been found in a G protein coupled receptor. Synthesis of metabolically labeled receptor in the presence of 1 mM phenyl-N-acetyl-alpha-D-galactosaminide, a competitive inhibitor of N-acetyl-alpha-D-galactose and N-acetylneuraminic acid transferases, also produced a receptor that migrated faster in denaturing gels. Serines and threonines present in the amino terminus were analyzed by alanine scanning mutagenesis to identify the acceptor sites. O-glycosylation was found at most serines and threonines present in the amino terminus. Because the disappearance of a site opened the availability of others to the transferases, the exact identification of the acceptor sites was not feasible. The wild type V2R expressed in HEK 293, COS, or MDCK cells underwent N- and O-linked glycosylation. The mutant V2R bearing all serine/threonine substitutions by alanine at the amino terminus yielded a receptor functionally indistinguishable from the wild type protein, whose mobility in polyacrylamide gels was no longer affected by sialidase treatment. PMID- 10362844 TI - A study of the intracellular and secreted forms of the MUC2 mucin from the PC/AA intestinal cell line. AB - In this study we present data on the entire population of MUC2 molecules secreted from and within the cell layer of an intestinal cell line. The molecular size distribution of the extracted molecules and their reactivity with two different MUC2 polypeptide antibodies indicated the presence of precursor and mature forms of the mucin. Oligomerized forms of the mucin were found in both the cell layer and medium; however, precursor forms were confined to the cell layer. Isopycnic density gradient centrifugation gave good resolution of mature and precursor forms of MUC2 as assessed by agarose gel electrophoresis. Three different populations of MUC2 were identified: one at low density (>1.3 g/ml) containing the N-glycosylated, non-O-glycosylated polypeptide; a second at intermediate density (1.3-1.35 g/ml) which may represent partially O-glycosylated intermediates; and a third at high density (1.36-1.48 g/ml) containing the mature MUC2 mucins. Rate-zonal centrifugation and agarose electrophoretic analysis of the low-density fraction indicated that the N-glycosylated MUC2 polypeptide was present as putative monomer and dimer/oligomer species. The combination of isopycnic density gradient centrifugation with agarose electrophoresis provides a new and simple approach that allows us to follow the MUC2 gene product from polypeptide through to the mature glycosylated mucin. PMID- 10362845 TI - The Joan Mott Prize Lecture. The integrated response to hypoxia: from circulation to cells. PMID- 10362846 TI - Effect of extracellular cations on the inward rectifying K+ channels Kir2.1 and Kir3.1/Kir3.4. AB - The effects of Ba2+, Mg2+, Ca2+ and Na+ as blocking ions were investigated in 90 and 10 mM extracellular K+ solutions on the cloned inward rectifying K+ channel Kir2.1 expressed in Xenopus oocytes. Some data were also obtained using another inward rectifying K+ channel Kir3.1/Kir3.4. The addition of Ba2+ caused a concentration-, voltage- and time-dependent block of both channels. Decreasing the extracellular K+ concentration augmented the block. The data suggest that Ba2+ blocks the channels by binding to a site within the channel pore and that the electrical binding distance, delta, of the site is significantly different for Kir2.1 and Kir3. 1/Kir3.4 (0.38 and 0.22, respectively). Mg2+ and Ca2+ caused an instantaneous concentration- and voltage-dependent block of both channels. With Kir2.1, decreasing the K+ concentration augmented the block. The voltage dependence of the block was less than that of Ba2+ ([delta], 0.1), indicating a more superficial binding site for these ions within the channel pore. The affinity of the channels for Mg2+ and Ca2+ was 1000-fold lower than that for Ba2+. Addition of Na+ resulted in a concentration-, voltage- and time-dependent block of Kir2.1, similar to that observed with Ba2+. The competition between the blocking cations (for Kir2.1: Ba2+, Mg2+, Ca2+; for Kir3. 1/Kir3.4: Ba2+) and extracellular K+ suggests that the binding sites for the blocking cations may be sites to which K+ binds as part of the normal passage of K+ through the channels. It is possible that under normal physiological conditions naturally occurring extracellular cations may partly block the two inward rectifying K+ channels. PMID- 10362847 TI - Activation of ionic channels by deoxycholate in frog and human cell lines. AB - Humans, after extensive ileal resection, frequently suffer from diarrhoea, which may be due to an increased delivery of deoxycholate (DOC) to the large intestine. In the frog skin the addition of DOC (0.5 mM) to the apical side induced the activation of amiloride-sensitive Na+ channels and an increase in the unidirectional Cl- fluxes. Here we used two established cell lines (A6 and Caco2) to study the effect of DOC on ion channels at cell and membrane level using the patch-clamp technique. In A6 cells subcultured directly on Petri dishes and studied in the whole-cell configuration, DOC induced an increase in cell conductance of 110.3 +/- 4 pS pF-1 (N = 8) which was reduced to 89 +/- 14 pS pF-1 (N = 8) by the addition of DIDS (0.5 mM), The absolute values of these two effects were not statistically different (P < 0.2). In Caco2 cells, the addition of DOC (0.5 mM) induced, after 1 min, an increase in cell conductance of 583 +/- 16 pS pF-1 (N = 8) which was reduced to 560.4 +/- 16 pS pF-1 (N = 8) by DIDS (0.5 mM) and N-phenylanthranilic acid (DPC; 0.5 mM). The two values were not statistically different (P < 0.4). In Caco2 cells subcultured under the same conditions, DOC induced an increase in cell conductance of 1710 +/- 64 pS pF-1 (N = 6). Subsequent addition of amiloride (0.1 mM) reduced the cell conductance to 1558 +/- 33 pS pF-1 (N = 6). These two mean values were statistically different allowing for an error of the second kind < 0. 05. In cells in which DOC produced a conductance increase of 1010 +/- 10 pS pF-1, gadolinium (0.5 mM) induced a fall in cell conductance of 1800 +/- 10 pS pF-1. In Caco2 cells, addition of DOC (0.5 mM) to the bath reversibly induced the appearance of or an increase in channel activity in patches studied in cell-attached and excised inside-out configuration. In inside-out experiments (N = 13) DOC (0. 5 mM) induced the appearance of channel activity with conductances and reversal potentials (Er) of 27.7 +/- 1.9 pS and 0.8 +/- 5.7 mV, respectively. In cell-attached patches (N = 13) these values were 24.9 +/- 4.4 pS and -18.1 +/- 6.4 mV. In excised inside-out patches from Caco2 cells, subjected to electrochemical gradients for Na+, K+ and Cl-, (+85, -85 and 0 mV, respectively), addition of DOC also induced an increase in the baseline conductance and a shift in the reversal potential from values around +25 mV to values around 0 mV. Bile salts activated both anionic and cationic channels and did not require the presence of intracellular factors for these effects. We suggest that they act at the membrane level. PMID- 10362848 TI - High activity K+ channels in rat hippocampal neurones maintained in culture. AB - A channel was identified in cell-attached recordings in rat hippocampal neurones maintained in culture. This channel, which was highly active at the resting membrane potential, was present in most (73 %) patches studied. The channel was characterized by long duration openings and a high open probability (Po, mean value 0.73 at -70 mV) at negative patch potentials with mild voltage dependence over the range -40 to -120 mV. It showed inward rectification. There were up to five active channels in cell-attached recordings in experiments where the cells were bathed in sodium-containing Locke solution. The single channel conductances in cell-attached recordings with 140 or 40 mM K+ in the patch pipette were 26 and 12 pS, respectively. The channel was therefore selective for K+ over Na+. The channel was not permeable to Rb+ ions. The single channel conductance was 24 pS in excised inside-out patches bathed in symmetrical K+ (140 mM) solutions. Examination of the channel kinetics revealed that both the open and closed time distributions could be fitted by the sum of three exponentials, there being no pronounced voltage sensitivity between -60 and -120 mV. The 26 pS K+ channel was insensitive to extracellular TEA, apamin, 4-AP and dequalinium. Neither was it sensitive to intracellular Ca2+. Extracellular Ba2+ was effective in reversibly blocking the channel, the IC50 being 2.0 mM. There was no obvious effect of bath application of the K+ channel opener, lemakalim, or a cAMP analogue. This channel appears to contribute a significant proportion (at least 30 %) of the resting conductance in these neurones. PMID- 10362849 TI - Chemoattractant- and mitogen-induced generation of reactive oxygen species in human lymphocytes: the role of calcium. AB - This study examined the role of calcium in the generation of reactive oxygen species (ROS) in human lymphocytes activated by the chemoattractant formyl-Met Leu-Phe (fMLP) and the T-cell mitogen phytohaemagglutinin (PHA). The concentrations of cytosolic calcium ([Ca2+]i) and ROS were monitored simultaneously with a fluorescence spectrophotometer after the cells had been incubated in fura-2 (calcium-sensitive dye) and 2',7'-dichlorofluorescein diacetate (DCF-DA, ROS-sensitive dye). The lymphocytes were stimulated with fMLP (200 nmol l-1) or PHA (10 micromol l-1) in the absence and presence of extracellular calcium. A dose-response test was also conducted for extracellular calcium. fMLP and PHA significantly increased both [Ca2+]i (P < 0.001) and ROS concentrations (P < 0. 001) above the control levels in the presence of extracellular calcium. However, such increases were abolished in the absence of extracellular calcium, suggesting total dependence of the responses to both fMLP and PHA on transplasma-membrane calcium influx. There were also graded increases in ROS with increasing concentrations of extracellular calcium. The results show that transplasma-membrane calcium influx is essential for fMLP- and PHA-induced generation of reactive oxygen species in human lymphocytes. PMID- 10362850 TI - The role of Na+-H+ exchange in fluid and solute transport in the rat efferent ducts. AB - In vivo microperfusion techniques were used to investigate the role of Na+-H+ exchange in the efferent ducts of the rat. Individual efferent ducts were perfused with a Krebs-Ringer bicarbonate solution (KRB) containing 0, 1, 3, 5 or 7.5 mM amiloride. Concentrations of 1-5 mM amiloride inhibited fluid reabsorption from the efferent ducts in a linear dose-dependent manner with an apparent Km of 3 mM. Inhibition was maximal at 5 mM with reabsorption reduced by about 70 %. The effects of amiloride were completely reversible and there was little effect of amiloride on luminal osmolality and concentrations of Na+, Cl- or K+. It is concluded that Na+-H+ exchange is one of the principal mechanisms responsible for fluid and electrolyte reabsorption in the efferent ducts and offers a means by which the efferent ducts are able to achieve flow-dependent, autoregulated fluid reabsorption. PMID- 10362851 TI - Effects of changes in pH and PCO2 on wall tension in isolated rat intrapulmonary arteries. AB - We examined mean ( S.E.M.) changes in wall tension in isolated rat intrapulmonary arteries on switching from control conditions (pH 7.38 +/- 0.01; PCO2, 34.4 +/- 0.5 mmHg) to hypercapnic acidosis (pH change, -0.24 +/- 0.01; PCO2 change, +27.5 +/- 0.9 mmHg), isohydric hypercapnia (pH change, -0.02 +/- 0.01; PCO2 change, +28.5 +/- 0.8 mmHg) and normocapnic acidosis (pH change, -0.24 +/- 0.01; PCO2 change, -0.5 +/- 0.3). Arteries were submaximally preconstricted with prostaglandin F2 and changes in tension are expressed as a percentage of the 80 mM KCl-induced contraction (%Po). Mean changes in wall tension on switching to hypercapnic acidosis (+4.4 +/- 3.7 %Po), isohydric hypercapnia (+1.9 +/- 2.2 %Po) and normocapnic acidosis (-1.5 +/- 1.9 %Po) were not significantly different from the change observed on switching to control conditions (+3.5 +/- 1.1 %Po), and were unaltered by endothelial removal. In isolated carotid preparations, the change in tension in isohydric hypercapnia (-6.8 +/- 7.1 %Po) was not significantly different from that observed in control switches (+8.6 +/- 3.2 %Po). Significant reductions in tension (P < 0.001) were observed in hypercapnic (-42.9 +/- 7.8 %Po) and normocapnic acidosis (-36.4 +/- 9.0 %Po). These data suggest that intrapulmonary arteries are resistant to the vasodilator effects of extracellular acidosis observed in systemic (carotid) vessels. PMID- 10362852 TI - Inhibition of mucosal glycylsarcosine uptake by acetate in rat distal small intestine. AB - Acetate deriving from microbial fermentation may occur at considerable concentrations in the distal small intestine, where it appears to be absorbed by two different mechanisms: acetate-HCO3- exchange and non-ionic diffusion. Whether acetate affects absorption of other nutrients at this intestinal site is not known. Therefore the influence of acetate (30 mmol l-1) on oligopeptide absorption was studied using an in vitro mucosal uptake technique allowing measurement of substrate uptake across the brush border membrane (BBM). Acetate significantly inhibited mucosal uptake of 14C-labelled glycylsarcosine (Gly-Sar) at pH 6 and pH 7 in the presence of sodium. No inhibition occurred in the absence of sodium. Both acetate and the absence of sodium decreased Vmax of mucosal Gly Sar uptake without substantially affecting the apparent Km value. Since it is well established that Vmax of peptide transport across the intestinal BBM depends on the size of the transmembrane H+ gradient as a driving force the present findings are in accordance with the assumption that acetate inhibits peptide absorption by attenuating the H+ gradient across the BBM, which depends on the presence of sodium. PMID- 10362853 TI - Insulin sensitivity, clearance and release in kininogen-deficient rats. AB - Insulin sensitivity of kininogen-deficient rats was compared with that of normal rats using euglycaemic hyperinsulinaemic glucose clamping. Anaesthetized animals were infused with 2-50 mU kg-1 min-1 of insulin and the glucose infusion rates needed to maintain euglycaemia were determined. Maximum glucose uptake, insulin sensitivity index and insulin clearance were reduced in kininogen-deficient rats. Captopril increased the amount of glucose needed to maintain euglycaemia during infusion of 2 and 10 mU kg-1 min-1 of insulin in normal rats, but had no effect in kininogen-deficient rats. Anaesthetized rats of both strains were given an intraperitoneal injection of glucose and the evolution of blood glucose was followed for 120 min. The peak increase was higher in kininogen-deficient rats. Similar larger increases in blood glucose were observed after glucose injection in normal rats previously treated with HOE 140, a bradykinin B2 receptor antagonist. After glucose injection, plasma insulin increased in both groups of rats but reached lower levels in kininogen-deficient animals. These results suggest that bradykinin is involved not only in the clearance of glucose and insulin by the tissues during insulin infusion but also that bradykinin can affect the release of insulin after a glucose load. PMID- 10362854 TI - Vagotomy suppresses cephalic phase insulin release in sheep. AB - The effect of selective vagotomy of the abomasum, pylorus, duodenum and liver on insulin release during the cephalic phase of digestion was investigated in wethers and lactating ewes. Electrical stimulation of the cervical vagus nerves was carried out to test the completeness of the vagotomies performed. In experiment 1, using wethers, the abomasal, pyloric and duodenal branches (ADV; n = 7) or the hepatic, abomasal, pyloric and duodenal branches (HADV; n = 10) of the ventral and/or dorsal vagus nerves were cut; a third group of wethers underwent sham-operation (SO; n = 8). In experiment 2, vagotomy (ADV; n = 5) or sham-operations (SO; n = 5) were carried out in lactating ewes. Jugular blood was drawn before and after presentation of food for glucose and insulin determination (experiments 1 and 2) or before, during and after the electrical stimulation of the peripheral ends of the cut cervical vagus nerves in randomly selected lactating ewes (experiment 3: ADV = 3, SO = 3) and wethers (experiment 4: ADV = 4, HADV = 4, SO = 4), for determination of insulin only. Presentation of food caused an immediate and significant (P < 0.05) rise in plasma insulin levels in SO animals compared with ADV or HADV wethers (experiment 1) or ADV ewes (experiment 2) without any significant change in blood glucose concentrations. In comparison with the SO group the baseline-corrected areas under the insulin response curve were significantly (P < 0.05) smaller for the respective vagotomized groups for periods 1-2, 2-4 and 4-6 min (experiment 1) and 1-2 and 2 4 min (experiment 2) after presentation of food. Total area under the response curve for 10 min was significantly (P < 0.05) lower (experiment 1) and tended (P < 0.10) to be lower (experiment 2) for the vagotomized groups compared with that of the control groups. Direct electrical stimulation of the cervical vagus nerves raised plasma insulin concentrations to significantly (P < 0.05) higher levels in the SO ewes but not in the ADV ewes (experiment 3). It was also evident that in experiment 1, HADV did not have any additive effect over that achieved by ADV alone. These results indicate that the vagal innervation of the gut mediates insulin release during the cephalic phase of feeding in sheep. It is concluded that insulin secretion from the pancreatic -cells in response to either food related reflex activation of the vagal nuclei in the hypothalamus or direct cervical vagus nerve stimulation is mediated through the vagal efferent fibres carried in the abomasal, pyloric and duodenal branches of the vagus nerves in sheep. PMID- 10362855 TI - Cardiovascular changes associated with dehydration and drinking in unrestrained, lactating goats. AB - The aim of this study was to investigate if the alertness connected with seeing water increased arterial blood pressure and heart rate to the same extent as the act of drinking, and if ingestion of warm water caused a different effect compared with ingestion of cool water on these cardiovascular variables. Seven goats of the Swedish domestic breed (Capra hircus) were used in a cross-over design. The animals were dehydrated for 24 h. They were allowed to watch water being prepared for 11-16 min, after which they were given access to warm (35 degrees C) or cool (15 degrees C) water. The goats drank 6.86 +/- 0.36 l of the warm water and 4.54 +/- 0.35 l of the cool water (P < 0.05) within the first hour. The arterial blood pressure, heart rate and activity of the animals were registered by an implanted telemetric device. Dehydration did not affect the cardiovascular variables, except before feeding in the morning, when the heart rate accelerated faster in dehydrated goats. Heart rate increased abruptly when dehydrated goats saw water being prepared, remained at the increased level during drinking and then slowly declined. It increased again during the afternoon feeding, to a level similar to that on control days, but between 18.00 and 06.00 h the heart rate was higher than during control nights. Blood pressure did not change when the goats saw water, but increased when they drank. On the morning following rehydration, the rise in heart rate in response to feeding was delayed compared with that during control and dehydration periods. It is concluded that seeing water caused arousal in the goats, resulting in an accelerated heart rate. The additional rise in blood pressure during the act of drinking appears to be a combination of excitement and sensory inputs from the pharyngeal region, causing a temporary activation of the sympathetic nervous system. PMID- 10362856 TI - Effects of variations in live weight gain on bone growth and composition and on markers of bone turnover in lambs. AB - Growing lambs were fed the same diet at intakes supporting mean live weight gains of 0.1, 0.2 and 0.3 kg day-1, representing slow, intermediate and fast growth groups, respectively. The effects on bone growth and composition, and on blood and urinary bone marker concentrations or excretion rates were monitored. Compared with the slow-growing lambs, the higher intake group grew twice as fast, had higher rates of bone growth (indicated by external metatarsal length), and larger and heavier bones at slaughter. Bones from fast-growing animals had higher collagen and deoxypyridinoline concentrations, and lower Ca:collagen, Ca :P and pyridinoline : deoxypyridinoline ratios, indicating a less mature bone compared with the slow-growing lambs. Bone growth rate had no effect on plasma osteocalcin, bone-specific alkaline phosphatase or growth hormone concentrations, nor on the urinary excretion of pyridinoline and deoxypyridinoline. The results for plasma markers may be explained by an increase in blood volume linked with increased body weight. PMID- 10362857 TI - Anaerobic power of the arms and legs of young and older men. AB - The purpose of this study was to examine differences in the anaerobic exercise performance of young and older men. Eight healthy, active older (68.5 +/- 2.4 years old, mean S.D.) and eight healthy, active young (30.6 +/- 4.5 years old) subjects were assessed for peak and mean power output (PP and MP, respectively) of the legs and arms, during 30 s Wingate tests. PP during leg exercise was significantly (P < 0.05) higher in the young (14.6 +/- 1.6 W kg-1) compared with the older (10.7 +/- 1.0 W kg-1) group. MP of the legs was also greater in the young subjects (10.7 +/- 0.7 vs. 7.4 +/- 0.9 W kg-1). These differences in PP and MP remained significant when expressed relative to lean leg volume. PP during arm cranking was significantly greater in the young subjects (8.9 +/- 0.7 vs. 7.5 +/- 0.6 W kg-1) as was MP (6.4 +/- 0.7 vs. 5.0 +/- 0.7 W kg-1). Post-exercise blood lactate concentration in the older group (7.0 +/- 1.6 mmol l-1) was less (P < 0.05) than in the young group (10.6 +/- 2.0 mmol l-1), for leg work only. The significant loss of anaerobic power in the older group could not be explained by a difference in muscle mass. Power output was also lower in the arms, but to a lesser extent. The results of this study suggest that a reduction in the ability to perform high intensity exercise may be an inevitable consequence of ageing. The extent, however, of this decline varies with different muscle groups. PMID- 10362858 TI - [Results of surgery of the endolymphatic sac]. AB - Surgery of the endolymphatic sac is controversial. Some consider it a placebo and others consider it the surgical treatment of choice in Meniere's disease. We studied the medical records of 87 patients who underwent surgery between 1978 and 1996. Simple decompression was practiced in 89% and a House shunt in 27.6%. The 1 year results were improvement or recovery from vertigo in 65.4%, no improvement in 25%, and reoperation in 9.5%. Tinnitus improved in only 11.9% and hearing loss improved in 9.5% and worsened in 19%. We reviewed the cases of 50 patients with a 5-year follow-up(60%). Vertigo improved in 72%,hearing loss worsened in 72%, and tinnitus remained unchanged in 78%. In view of the good results, scant complications and simplicity of the surgical procedure, we consider endolymphatic saccule surgery to be the first choice in Meniere's disease that is unresponsive to medical treatment. PMID- 10362859 TI - [Stapedectomy. Study of variables with special interest for the surgical technique]. AB - A study was made of the variables that influence primary and secondary stapedectomy. We studied stapedectomies performed in the last 5 years at our hospital. The only variable that significantly influenced outcome was technique. PMID- 10362860 TI - [Prognostic value of the stapedius muscle and electroneurography in facial paralysis a frigore, or Bell's palsy]. AB - A retrospective study was made of 170 patients diagnosed as Bell's palsy in a 7 year period (August 1990 to April 1997). We evaluated two prognostic factors: stapedial reflex and electroneurography findings. The condition was classified by nerve affection into three types: less than 10%, 10 to 30%, and over 30%. The evolution of our patients was: complete recovery in 86.4%, slight impairment in 2.9%, major impairment in 3.5%, and unknown in 7%. The stapedial reflex was present in 87 patients, absent then present in 26 patients, and absent in 5 patients. Electroneurography revealed less than 10% nerve involvement in 5 patients, 10-30% in 18 patients, and over 30% in 63 patients. PMID- 10362861 TI - [Bimastoid leads and bilateral stimulation in the recording of auditory-evoked brain-stem potentials]. AB - The recording of auditory brain stem potentials (ABR) with bimastoid leads and bilateral stimulation makes it possible to evaluate relative cochlear function and the type of hearing loss. The theoretical background for this technique and the results obtained in patients with disease of the internal-middle ear, cochlea, and auditory nerve are discussed. PMID- 10362862 TI - [Rendu-Osler disease. Follow-up of 6 patients treated by embolization. Nasal telecobaltotherapy in a refractory case]. AB - Patients with Rendu-Osler-Weber disease can present severe nose bleeding. The treatment of 6 patients with this condition is reported. Supraselective embolization was the treatment of choice. One patient showed no improvement with embolization and suffered massive nosebleed requiring blood transfusion. After a review of the literature, the nasal cavity was irradiated.s. PMID- 10362863 TI - [Study by S-100 polyclonal antibodies of Langerhans cells in vocal cord polyps]. AB - In 10 patients (5 smokers) with benign laryngeal polyp of a single vocal cord, a biopsy was performed. Five normal control specimens were obtained from the larynx of recently deceased patients who underwent postmortem study. Langerhans cells were counted in specimens of histologically normal and pathological vocal cord mucosa after identification with S-100 polyclonal antibody. Langerhans cells were 11.5-fold more common in the epithelium of vocal cord polyps than in the mucosa of normal vocal cords. PMID- 10362864 TI - [Hypothyroidism in patients treated for laryngeal cancer: preliminary results]. AB - OBJECTIVE: The reported incidence of hypothyroidism following surgery and/or irradiation for laryngeal cancer varies widely and the condition often is misdiagnosed. This study examines the incidence of thyroid dysfunction in patients with laryngeal cancer. MATERIAL AND METHODS: Thyroid function tests were carried out in 75 patients with stage III and IV laryngeal carcinoma who were treated in our center with surgery (13 cases), radiotherapy (13 cases), or surgery and radiotherapy (49 cases) at least 18 months earlier. The clinical and histological variables recorded included T4 and TSH concentrations. Univariate and multivariate analysis was carried out with the BMDP program from UCLA (1995 version) to examine the relationship between hypothyroidism and clinical and pathological factors. RESULTS: Twenty-nine patients (38.6%) had high TSH or low T4 concentrations and were diagnosed as having hypothyroidism. Hypothyroidism was significantly related with date of surgery (before 1993) and the treatment used on the neck. Thyroid function was rarely affected in patients who underwent functional neck dissection, but radical neck dissection and irradiation of the neck always were followed by hypothyroidism. CONCLUSIONS: Thyroid testing should be performed routinely in the follow-up of laryngeal cancer. Many psychological symptoms attributed to total laryngectomy may be due to hypothyroidism, an easily treated condition. PMID- 10362865 TI - [Rhabdomyosarcoma of the neck in adults]. AB - The clinical case of a 59-year-old male with neck rhabdomyosarcoma is reported. The literature is reviewed for the epidemiology, diagnosis, prognosis, and treatment of this infrequent sarcoma. PMID- 10362866 TI - [Postoperative complication in stapedectomy: excessive introduction of the prothesis in the oval window]. AB - Postoperative failures and complications detected in patients who undergo surgery for otosclerotic disease are not uncommon in stapes surgery. Prosthesis displacement and incus necrosis are the most common findings in review stapedectomy. We report the case of a patient who had tinnitus, vertigo, and non recovery of air conduction thresholds without neurosensorial lesions after stapes surgery. The suspected diagnosis of excessive introduction of the prosthesis in the oval window was confirmed by computed tomography, which showed the radio opaque image of the McGee metal prosthesis. The prosthesis replacement and literature review are discussed. PMID- 10362867 TI - [Gushers. Perilymphatic leaks]. AB - We report an unusual complication of stapedectomy called <>. It was resolved and the patient's hearing level improved without vertigo. PMID- 10362868 TI - [Otorhinolaryngological manifestations of varicella-zoster virus]. AB - Otological complications of varicella-zoster syndrome (Ramsay Hunt syndrome) include facial paralysis, tinnitus, hearing loss, vertigo, dysgeusia, and skin rash. The lower cranial nerves sometimes are affected by this neuritis. A case is reported of a woman without immune-system impairment who had cranial multineuritis with unilateral involvement of the VII, VIII, IX and X cranial nerves after infection with varicella-zoster virus without herpetic lesions. PMID- 10362869 TI - [Secondary brain herniation. A case report]. AB - A 47-year-old woman with left temporal bone herniation had a history of treatment of chronic cholesteatomatous otitis media. After clinical and radiological diagnosis, the defect was repaired using a middle fossa craniotomy approach and a Vicryl-collagen(R) plate. There were no postoperative surgical complications and the repair was successful. The sequellae of herniation of the temporal lobe and dura are potentially lethal and difficult to correct surgically. PMID- 10362870 TI - [Heterotopic brain tissue in the pharynx]. AB - The presence of heterotopic brain tissue in the head and neck region is exceptional. Most reported cases have been observed in the nasal region and are called nasal gliomas. The case of a 10-month-old boy with heterotopic brain tissue on the soft palate and nasopharynx is reported. Surgical treatment was successful and no complications or recurrences have been observed in 7 years of follow-up. The main pathogenic theories and nomenclature are analyzed. PMID- 10362871 TI - [Inflammatory pseudotumor of the larynx]. AB - Inflammatory pseudotumors are rare and usually located in lung, although they can develop in any organ. They are unusual in head and neck and very rare in the larynx. A case of laryngeal pseudotumor in a patient with odinophagia and neck pain is reported. After diagnosis, the patient was treated with steroids, which produced complete resolution. These lesions can simulate malignant neoplasms and must be considered in the differential diagnosis. PMID- 10362872 TI - [Recurrential paralysis as the first manifestation of chordoma of the petrous apex]. AB - We report the case of a 67-year-old woman with paralysis of the right recurrent nerve. Imaging studies showed a mass in the right petrous apex and jugular foramen. Surgery was performed using a lateral approach to the skull base. The tumor excised from the petrous apex was a chondroid chordoma. Chordomas are rare dysontogenic neoplasms that arise from notochord remnants, generally on the midline. It should be included in the differential diagnosis of tumors of the skull base. PMID- 10362873 TI - [Hyalinizing trabecular adenoma of the thyroid]. AB - Hyalinizing trabecular adenoma (HTA) is a follicle-derived tumor of the thyroid gland that has been described recently. The differential diagnosis includes medullar carcinoma, papillary carcinoma, and paraganglioma of the thyroid. We report a case of thyroid HTA in a 25-year-old woman. The tumor appeared as a <> of the left thyroid in radionuclide scans. Histologically, the tumor showed tumor cells arranged in trabeculae and a prominent hyaline stroma. The neoplastic cells were focally immunoreactive for thyroglobulin and negative for calcitonin, chromogranin, and S-100 protein. Three years after hemithyroidectomy, the patient is alive and free of disease. PMID- 10362874 TI - [Detection of fecal occult blood and colorectal cancer]. PMID- 10362876 TI - Iron-deficiency anemia due to chronic gastrointestinal bleeding. AB - AIMS: chronic gastrointestinal bleeding is the most common cause of iron deficiency anemia (IDA) in the general population. The objectives of this study were to determine the most frequent gastrointestinal lesions in IDA, the frequency and localization of potentially bleeding lesions, the value of the clinical history in diagnosis, the value of fecal occult blood testing, and the most appropriate diagnostic procedure for these patients. METHODS: we prospectively studied 80 patients older than 40 years with IDA, using upper gastrointestinal tract (GI) endoscopy and colonoscopy, beginning with the former (group A) or the latter (group B) depending on the clinical findings. Barium enema was done when colonoscopy was incomplete or unsatisfactory. If all these tests were negative, conventional barium contrast study of the small intestine and arteriography were done, if necessary. RESULTS: upper GI endoscopy found at least one lesion in 50 patients (72%), 13 in association with a colonic lesion (26%). Colonoscopy detected at least one lesion in 31 patients (45%), among whom 11 had another upper GI lesion (35.5%). Barium enema was positive in 4 out of 24 patients (17%). Barium contrast study of the small intestine detected lesions in 1 out of 7 patients (14%), and arteriography in 1 out of 4 patients (25%). The most common upper GI lesions were of peptic origin (esophagitis in 10, gastroduodenal erosions in 10, and peptic ulcer in 8). Neoplasms (17 cancers and 3 polyps) were the most common colonic lesion. Thirteen out of 38 patients (34%) with a potentially bleeding benign upper GI lesion had another lesion in the colon. The fecal occult blood test was positive in 9 out of 10 patients with colonic cancer and in 5 out of 9 with gastric cancer (74% positive predictive value). Nonsteroid antiinflammatory drug use did not correlate with the presence, location or type of lesion. The reliability of the clinically suspected origin of bleeding was 96% sensitivity, 43% specificity and 74% positive predictive value in group A, and 34%, 93% and 80% respectively in group B. CONCLUSIONS: lesions that cause chronic bleeding were more frequently located in the upper digestive tract than in the colon. There was a high prevalence of neoplasms in patients with IDA. One-third of the patients with a potentially bleeding benign lesion in the upper digestive tract had another lesion in the colon. A positive fecal occult blood test correlated highly with neoplastic lesions, and the presence of blood in the stool did not indicate whether bleeding originated in the upper or lower GI tract. Clinical history was of limited value in predicting the location of a bleeding lesion, but can be suggestive of a prior upper GI tract exploration. These patients need a complete study of both the upper and lower GI tracts. In patients in whom the aforementioned explorations are negative, the small bowel should be studied. PMID- 10362875 TI - Colorectal cancer screening through detection of fecal occult blood in a controlled health zone. AB - AIM: to determine the viability and utility of fecal occult blood (FOB) screening (Hemoccult II) as a method for the early diagnosis of colorectal cancer in a well controlled health area. METHODS: between February 1994 and September 1996, personal letters were sent to all persons in the Casas Ibanez health area (Albacete) aged between 50 and 75 years (4986 persons). All the tests were read by the same team member. A clinical history was taken of persons with positive test results, and they were offered colonoscopy. If a growth was found which was suitable for endoscopic resection, this was done. If the growth was totally resected an annual check-up was advised. Malignant and partially resected premalignant growths were referred for surgical treatment. We analyzed percent participation, attendance for testing, negative and positive tests, growths diagnosed, false-positive tests and positive predictive value. RESULTS: the a participation rate was 56.25%. In the 157 colonoscopies performed, 39 neoplasic polyps and 9 cancers were found (7 of which were Dukes stage A and 2 of which were Dukes stage C-D). Sensitivity was 97% and specificity was 96%. The positive predictive value was 30.37%. The positivity rate was 6.08%, while the predictive value for cancer was 5.38% and the predictive value for adenoma was 23.35%. CONCLUSIONS: FOB screening, in our setting, proved to have a high recruitment capacity with a positive predictive value for colorectal carcinoma which was slightly higher than the value obtained in other countries of similar socioeconomic status. Furthermore, we found a significant improvement in the diagnostic stage of colorectal carcinoma at the time of resection. PMID- 10362877 TI - Laparoscopic cholecystectomy: analysis of risk factors for predicting conversion to open cholecystectomy. AB - AIM: to assess the usefulness of different clinical and ultrasound parameters to identify patients at high risk of conversion from laparoscopic to open cholecystectomy. METHODS: we retrospectively reviewed the clinical records and preoperative ultrasonographic images of 80 patients who underwent laparoscopic cholecystectomy. RESULTS: eight clinical and nine ultrasound parameters were assessed. Our statistical analysis showed that ultrasound imaging indicated two risk factors for conversion from laparoscopic to open cholecystectomy: a scleroatrophic gallbladder and dilation of the intrahepatic biliary ducts. A gallbladder wall thicker than 6 mm was also considered a risk factor, although this difference was not statistically significant. CONCLUSIONS: our results suggest that preoperative ultrasonography is useful in selecting patients who are highly likely to require conversion from laparoscopic to open surgery. PMID- 10362878 TI - Hepatic cytochrome oxidase in rats with microsurgical cholestasis or portocaval shunt. AB - AIMS: portocaval shunt and extrahepatic cholestasis are experimental models of chronic hepatic insufficiency of different etiology and histological characteristics, and which probably also differ in the mechanism of impairment of oxidative metabolism. To test this hypothesis we measured hepatic cytochrome oxidase. METHODS: cytochrome oxidase was assayed with a histochemical technique in three groups of Wistar rats: A (n = 8) control; B (n = 8) microsurgical extrahepatic cholestasis; and C (n = 8) end-to-side portocaval shunt. RESULTS: cytochrome oxidase activity was lowest in group B, both in the left middle (p = 0.00019) and in the inferior caudate (p = 0.00014) hepatic lobes, and was highest in group C in both hepatic lobes, especially in the left middle lobe (p = 0.0029). CONCLUSION: the decrease in cytochrome oxidase activity in the liver of rats with extrahepatic cholestasis and the increase in animals subjected to portal flow deprivation demonstrate the different nature of the impairment in hepatic oxidative metabolism in these two pathological conditions. PMID- 10362879 TI - [Abdominosacral resection for locoregional recurrence rectal cancer]. AB - Local recurrence of rectal cancer occurs in up to 30% after radical surgical treatment and it represents a formidable challenge to surgeons and oncologist, presenting most of times within two years after proper therapy have been provided. Although chemoradiation therapy reduces the rate of it, it has no any impact in survival. On the other hand, it has been proved that almost 50% of recurrences are without evidence of systemic disease and amenable to surgical resection, by the time of diagnose. For this reason there are a number of authors currently arguing a more agressive treatment for this entity in order to improve survival and reduces recurrence rate. Radical pelvic surgery for recurrent rectal cancer should be performed primarily with curative intent in patients without evidence of extrapelvic or distant spread. Abdominosacral resection represents a therapeutic option for patients with specific type of pelvic recurrence providing, according to figures from the most experienced groups, an actuarial survival rates of almost 33% at four years in a group of patients with a life expectancy, by other means, round seven months. We present our experience with this surgical procedure in Surgical Oncology Department at Roger Williams Cancer Center in Providence, leads by HJ. Wanebo. PMID- 10362880 TI - [Metallic stents in recurrent benign biliary stricture]. PMID- 10362881 TI - [Cystoadenocarcinoma of the biliary tree: an uncommon malignant tumor]. PMID- 10362882 TI - [The importance of angiography of the surgical resection piece in the diagnosis of angiodysplastic lesions]. PMID- 10362883 TI - [Subcutaneous emphysema as postoperative complication of a colostomy closure]. PMID- 10362884 TI - Measuring postural changes in blood pressure in the healthy elderly. AB - BACKGROUND: Background The reproducibility of postural changes in blood pressure of a healthy elderly population determined using standard clinical measurements is not known. OBJECTIVE: To assess the differences in reproducibility of postural changes in blood pressure in healthy elderly subjects 1 and 3 min after standing within a day and between visits spaced 6 weeks apart. METHODS: Casual readings of blood pressures of supine and standing subjects were measured twice during the day by the same observer on two occasions 6 weeks apart using a semi-automatic syphgmomanometer. Twenty-two subjects with no known risk factors for orthostatic hypotension (13 men) aged 69+/-3 years (mean+/-SD) with a mean initial screening supine blood pressure of 153+/-19/88+/-11 mmHg were recruited. RESULTS: There were significant differences(P<0.001) between the postural changes both for systolic and for diastolic blood pressure between 1 and 3 min of standing, the largest falls occurring after 1 min of standing, though we found no variation between morning and afternoon measurements and between visits. The coefficients of reproducibility between visits for the postural changes in blood pressure after 1 and 3 min of standing were large both for systolic and for diastolic blood pressure, ranging from 9.8 to 29.3 mmHg. CONCLUSIONS: There was a consistent and significant difference between the postural changes in blood pressure after 1 and 3 min of standing for this healthy elderly population, but there was no significant difference between the postural changes in morning and afternoon measurements and between visits. This marked variability in the postural change in blood pressure with duration of standing must be taken into account when assessing the prevalence of orthostatic hypotension and the effects of treatment in patients with orthostatic falls in blood pressure. PMID- 10362885 TI - Individualized versus standardized analysis of ambulatory blood pressure profile: relationship with left ventricular characteristics. AB - OBJECTIVE: Objective To determine whether the use of patients' individual awake/asleep patterns instead of fixed day/night intervals would influence the correlations between blood pressure values and left ventricular morpho-functional characteristics. METHODS: We enrolled 167 never-treated hypertensives (clinic blood pressures >160 mmHg systolic or 90 mmHg diastolic, or both): 32 had 24h blood pressures <130/80 mmHg [white-coat hypertensives (WCH)] and 135 had 24h blood pressures >130 mmHg systolic or 80 mmHg diastolic, or both (hypertensives). Each patient underwent left ventricular echocardiographic examination and 24h ambulatory blood pressure monitoring, evaluated twice, using standard day/night intervals (daytime 0700-2200 h, night-time 2200-0700 h) and using the patient's individual awake/asleep pattern (an individualized scheme). RESULTS: Daytime and night-time blood pressures in WCH and daytime and night-time diastolic blood pressures in hypertensives were not affected by choice of using individualized or standard intervals; daytime systolic blood pressure in hypertensives was significantly higher and night-time systolic blood pressure lower with individualized intervals. The non-dippers (nocturnal decrease in blood pressure <10% of daytime blood pressure) were 31 hypertensives and six WCH with standard day/night intervals and 25 hypertensives and four WCH with individualized intervals; nocturnal falls in systolic and diastolic blood pressures were significantly greater with individualized intervals for both groups. Left ventricular hypertrophy was present in 68 hypertensives and seven WCH; left ventricular systolic function was normal in all and left ventricular diastolic function was impaired in 53 hypertensives and seven WCH. Left ventricular characteristics of WCH were not correlated to blood pressure parameters; left ventricular mass index of hypertensives was directly correlated to 24h, daytime and night-time systolic blood pressures, whereas left ventricular diastolic function was inversely correlated to night-time systolic and diastolic blood pressures. The correlations were not affected by choice of using individual awake/asleep patterns. CONCLUSIONS: Timing day and night in an individualized way seems to improve the evaluation of nocturnal fall in blood pressure, but does not improve the ability to predict the left ventricle's involvement with ambulatory blood pressure monitoring. PMID- 10362886 TI - Can structural equation models inform blood pressure research? AB - OBJECTIVE: To show how structural equation models might be used to better understand the ways in which risk factors influence blood pressure. METHODS: Nine measurements on 2009 women and 1518 men for whom there was complete data both at time 1 and at time 8 of the Framingham Heart Study were used to test a hypothetical model of how risk factors such as age, obesity, smoking, vital capacity, and heart rate influence each other and blood pressure. The hypothetical model was translated into structural equations and tested against the data. RESULTS: The hypothetical model fits the data for women at time 1 very well with a chi2=15.41 which, with 14 degrees of freedom, has P=0.32 and indicates there is no difference between the covariance structure generated by the hypothetical model and the covariance structure generated by the data. The same model was tested for women at time 8 and for men at times 1 and 8 also and fit almost as well. Age and percentage of ideal weight of subjects exert the strongest influence on systolic blood pressure, whereas the effect of age on diastolic blood pressure seems less consistent. Smoking has no direct effect on blood pressure, but it does have a small effect on heart rate and a negative effect on obesity, suggesting, perhaps, that, while it has no direct effect, it does play an indirect role. CONCLUSIONS: Structural equation models can be used by researchers trying to understand how risk factors can influence blood pressure in complex ways. The methodology is especially appropriate for testing competing conceptual models. PMID- 10362887 TI - Validation of two devices for self-measurement of blood pressure by elderly patients according to the revised British Hypertension Society protocol: the Omron HEM-722C and HEM-735C. AB - BACKGROUND: The validation of self-measurement devices for clinical use by elderly patients has been recommended. The Omron HEM-722C device has recently been validated according to the British Hypertension Society (BHS) protocol for use for general populations and the Omron HEM-735C is a new fully automatic device with a high capacity for storage of measurements that is integrated with a personal computer. OBJECTIVE: To perform a clinical validation for use by elderly people of the Omron HEM-722C and HEM-735C devices according to the revised protocol of the BHS and the criteria of the Association for the Advancement of Medical Instrumentation (AAMI). METHODS: We carried out a main validation test according to the revised BHS protocol for validation procedures for special groups on two groups of 30 subjects aged more than 65 years (29 men and 31 women), 11 of 30 with systolic blood pressures (SBP) <110mmHg, 10 of 30 with SBP >200 mmHg, 15 of 30 with diastolic blood pressures (DBP( <70 mmHg and 10 of 30 with DBP >110 mmHg. The results were graded according to the BHS system from A to D. RESULTS: The Omron HEM 722C achieved an overall A/A grading and satisfied the AAMI criteria for accuracy whereas the Omron HEM-735C achieved an overall B/A grading and satisfied the AAMI criteria for accuracy. The sphygmomanometer measurements were 147 +/- 31/79 +/- 15 and 144 +/- 30/78 +/- 15 mmHg (means+/-SD) respectively, for the models 722C and 735C. The average differences between mercury sphygmomanometer and HEM-722C readings for SBP and DBP were, respectively, 0.76+/-5 and 0.41+/-8 mmHg; those for HEM-735C were, respectively, 0.24+/-8 and 0.9+/-8 mmHg. Readings of the HEM-722C device differed by less than 5 mmHg for 76% of systolic readings and 96% of the readings differed by less than 10 mmHg. Diastolic measurements differed by less than 5 mmHg for 71% and less than 10 mmHg for 71 and 87% of all readings. Readings of the HEM-735C device differed by less than 5 mmHg for 68% of systolic readings and 74% of the readings differed by less than 10 mmHg. Diastolic measurements differed by less than 5 mmHg in 74% and less than 10 mmHg in 88 and 87% of all readings. CONCLUSIONS: On the basis of these results, for elderly subjects both self-measurement devices (Omron HEM-722C and HEM-735C) satisfied the validation criteria of the BHS and therefore can be recommended for the clinical measurement of blood pressure in elderly patients. PMID- 10362888 TI - Assessment of the performances of three oscillometric blood pressure monitors for neonates using a simulator. AB - BACKGROUND: The majority of monitors for non-invasive measurement of blood pressure in neonates and infants use the oscillometric method. OBJECTIVE: To use a simulator to investigate the overall system accuracy for three oscillometric blood pressure monitors. The tested devices are DINAMAP 8100 (Critikon), SpaceLabs M90426 (SpaceLabs Medical)and the module HP M 1008B (Hewlett-Packard). METHODS: The blood pressure values obtained by the three devices were compared with those of the invasive reference. A blood pressure simulator was used for testing the performance of the three blood pressure monitors. RESULTS: The Dinamap and SpaceLabs readings are generally in good agreement with the invasive reference. In contrast, the HP M 1008B readings for the diastolic and mean arterial pressures are inaccurate (the mean errors are 21 and 15mmHg, respectively). CONCLUSION: These results contradict our previously published data obtained when a different simulator (Cufflink)was used. These findings are important, because blood pressure values that are too low can result in a different therapeutic approach being applied, for newborn babies have only a small range of mean arterial pressure for the autoregulation of the cerebral blood flow. PMID- 10362889 TI - Evaluation of the Schiller BR-102 ambulatory blood pressure system according to the protocols of the British Hypertension Society and the Association for the Advancement of Medical Instrumentation. AB - OBJECTIVE: To evaluate the Schiller BR-102 monitor for ambulatory blood pressure measurement according to the protocols of the British Hypertension Society (BHS) and the Association for the Advancement of Medical Instrumentation (AAMI). DESIGN: The BHS protocol is divided into two parts. Part I, which is the part applicable to this study, comprises the main validation procedure and has five phases: before-use device calibration; in-use (field) phase; after-use device calibration; static device validation; and report of evaluation. METHOD: Three Schiller BR-102 recorders passed the before-use device calibration test, after which they entered the in-use (field) assessment phase during which the three recorders were each worn by 10 subjects for 24 h, after which calibration was again assessed. Because there was no difference in results of calibration testing among the three devices, one was selected randomly and the main validation test was carried out on 85 subjects with a wide range of blood pressures both for the auscultatory mode and for the oscillometric mode using the Sphygmocorder. The results were analysed according to the BHS grading system from A to D. The data were also analysed according to the standard of the Association for the Advancement of Medical Instrumentation (AAMI), which stipulates that the mean difference between the test device and the standard shall be 160/100 mmHg) the Schiller BR 102 was less accurate in the high pressure range for diastolic blood pressure but more accurate for systolic blood pressure, achieving A/C grading, while satisfying the AAMI criteria both for systolic and for diastolic blood pressure in the auscultatory mode. In the oscillometric mode the device performed less accurately in the high-pressure range, achieving grade D/C, while failing to satisfy the AAMI criteria both for systolic and for diastolic blood pressure. The means+/-SD of the first mercury sphygmomanometer measurements were 143+/-32 mmHg for systolic blood pressure and 88+/-21 mmHg for diastolic blood pressure. Acceptability to subjects was good and the manufacturer's manual was satisfactory. CONCLUSION: On the basis of these results, the Schiller BR-102 can be recommended for ambulatory blood pressure measurement in clinical practice using the auscultatory mode, but the oscillometric mode, which operates only if the device fails in the auscultatory mode, does not provide accurate measurements. PMID- 10362890 TI - Determination of accuracies of 10 models of home blood pressure monitors using an oscillometric simulator. AB - BACKGROUND: Patients and doctors often use home blood pressure monitoring (HBPM) to assess the control of hypertension. Despite its popularity there has always been some uncertainty with regard to its accuracy, reliability, reproducibility, and comparability. Although there are pre-market HBPM standards of accuracy, there are no standards to assure accuracy of individual HBPM units after they have been brought home. OBJECTIVE: Determination of reliability, reproducibility, and comparability of 10 models of home blood pressure monitors. METHODS: We used a Biotek BP Pump as an oscillometric simulator of systolic and diastolic blood pressures to determine reliability, reproducibility, and comparability of 10 devices. RESULTS: All of the units tested, except the Pollonex BP1000, produced reproducible readings with the pooled SD of four blood pressure settings less than 3.10 mmHg both for systolic and for diastolic measurements. The oscillometric blood pressure pump method was found to be very reproducible, with pooled differences of less than 2 mmHg and SD of less than 0.5 mmHg for a repeated series of measurements using the same monitor. Different machines of the same model were also very comparable, with pooled differences of less than 3.6 mmHg and pooled SD less than 0.7 mmHg both for systolic and for diastolic readings. There were 11-14 mmHg differences between models for all of the simulated blood pressure readings except that a 27 mmHg difference was measured at the 200 mmHg systolic blood pressure level. These differences will not necessarily be the same for measurements with humans instead of oscillometric signal generation. A system for grading the accuracy of the tested HBPM that defines accuracy of HBPM as within +/-2 SD of the average of 85 measurements is described. CONCLUSION: All models of home blood pressure monitors tested, with the exceptions of the Pollonex BP1000, produced reproducible readings and different machines of the same model were comparable. PMID- 10362891 TI - Modification of blood-pressure-measuring devices and the protocol of the british hypertension society AB - The market for blood-pressure-measuring devices is enormous and growing. Devices are required in a number of areas in hospitals, in doctors' offices, in paramedical areas and in the transport of patients; and there is a vast demand for self-measuring devices from the public. In Germany, for example, 12 million such devices are sold annually [1]. Only a fraction of the many hundreds of models available worldwide has been subjected to independent validation, though the number of devi E> 1359-5237 4 1 1999 February 1999 Paper alert 55 56 EN George A Mansoor Section of Hypertension and Clinical Pharmacology, University of Connecticut Health Center, Farmington, Connecticut, USA EDITORIAL http://www.cardiosite.com http://www. cardiosite.com PMID- 10362892 TI - Neurology in the electronic information age. AB - This review discusses the state of neurology and the Internet at the turn of the millennium. First, some basic definitions about the Internet and its component protocols are presented. Next, ways neurologists and patients can use the Internet are enumerated. Internet resources or applications are available or are being created that can aid in the successful fulfillment of a neurologist's core professional activities: clinical care, teaching, research, and practice issues. Currently, the most useful categories of Internet resources for neurologists are electronic communication and access to knowledge bases. They fulfill needs that are not met by traditional, non-electronic media. There are many other types of Internet applications that supplement traditional medical methodologies. Finally, some problems and prospects concerning medical uses of the Internet are discussed: technological infrastructure including usability, security, meaning, validity/quality, value, outcomes, and responsibility. These issues must be successfully addressed if Internet computing is to become truly useful 'just in time' at the point of medical care. Solutions are actively under development today. The prospects are bright for neurology, and medicine in general, on the Internet. The Internet will become an essential medical device in the near future. PMID- 10362893 TI - Apolipoprotein E phenotypes in demented and cognitively impaired patients with and without cerebrovascular disease. AB - Controversy exists regarding the apolipoprotein E (ApoE) epsilon4 allele association with vascular dementia (VaD), ranging from increased epsilon4 frequency, similar to that found for Alzheimer's disease (AD), to no association between the epsilon4 allele and VaD. To clarify further the relationship between ApoE alleles polymorphism and cerebrovascular disease (CVD) in demented and cognitively impaired patients, we examined the ApoE phenotypes in a sample of 280 patients: 155 with AD, 21 with VaD, 32 with mixed dementia (MD), 45 with mild cognitive impairment (MCI) but without CVD, and 27 in which vascular disease was the most probable cause of cognitive decline [vascular mild cognitive impairment (VMCI)]. Our results show that the frequency of the ApoE epsilon4 allele in patients over 70 years old with clinically diagnosed VaD and VMCI does not differ significantly from that of controls. In contrast, ApoE epsilon4 allele-bearing individuals had greater risk of having late-onset AD (OR = 8.8; 95% CI 3.7-21.0), or non-vascular cognitive impairment (OR = 7.0; 95% CI 2.5-19.0). PMID- 10362894 TI - A phase II study in patients with Alzheimer's disease to assess the preliminary efficacy and maximum tolerated dose of rivastigmine (Exelon). AB - Rivastigmine is a carbamate acetylcholinesterase (AChE) inhibitor with central selectivity. Early studies showed that daily doses up to 6 mg/day have some efficacy in patients with dementia of the Alzheimer type (DAT). The present study was designed to assess the safety, tolerability and efficacy of rivastigmine at doses up to 12 mg/day. A total of 114 patients with mild-moderate DAT were randomly assigned to either rivastigmine (b.i.d. (twice daily) or t.i.d. (three times daily)) or placebo in a double-blind fashion titrated to their maximum tolerated dose over 10 weeks followed by an eight-week maintenance phase. The mean maximum tolerated dose was approximately 10 mg/day (b.i.d. or t.i.d.). Gastrointestinal complaints, the majority of which were mild to moderate, were the most frequently reported adverse events. No clinically relevant changes in vital signs, haematology or organ function were detected. Significantly more patients taking rivastigmine b.i.d. were considered improved according to the Clinicians' Interview-Based Impression of Change-Plus (CIBIC-Plus) vs. placebo (57% vs. 16%, respectively; P = 0.027). The Nurses' Observation Scale for Geriatric Patients (NOSGER) (memory component) and the Alzheimer's Disease Assessment Scale-cognitive subscale (ADAS-cog) also improved in the rivastigmine b.i.d. group vs. placebo (mean change from baseline on NOSGER = -0.7 vs. +1.3, respectively; P = 0.037: mean change from baseline on ADAS-cog = -2.7 vs. +0.2, respectively; P = 0.054). Despite the relatively small size and limited duration of the study, the finding that rivastigmine induced changes in the same (positive) direction in all three dimensions measured suggests that rivastigmine at doses of up to 12 mg/day has useful efficacy in patients with mild-moderate DAT. Reports from larger phase III studies confirm this finding. The results of this study also suggest that b.i.d. is the more efficacious regimen and has comparable tolerability to the t.i.d. regimen. PMID- 10362895 TI - Positive association between an estrogen receptor gene polymorphism and Parkinson's disease with dementia. AB - Parkinson's disease (PD) is a common cause of dementia in the elderly. Dementia in Alzheimer's disease (AD) and PD share common biologic and clinical features. The estrogen receptor (ER) gene is one of the susceptibility genes for AD. In order to test the hypothesis that the overlap between AD and PD may have a genetic basis, we determined ER gene polymorphisms in 13 PD patients with dementia (PDD) (age +/- SD; 71.9 +/- 5.5 years), 71 PD patients without dementia (PDND) (68.4 +/- 7.5 years), 86 AD patients (76.8 +/- 8.0 years) and 51 control subjects (CTL) (74.9 +/- 6.9 years). ER genotypes were classified as a P or p allele on the basis of a Pvu II-RFLP, and X and x on the basis of a Xba I-RFLP. The frequency of the P allele in the PDD group as well as the AD group was higher than that in CTL. There was no significant difference in the distribution of the P allele between CTL and PDND. There were no significant differences in the distribution of the X allele among the PDD, PDND and CTL groups, whereas a higher incidence was found in AD. We conclude that the ER gene may be a common susceptibility gene for dementia in PD as well as AD. PMID- 10362896 TI - High levels of IL-10 secreting cells are present in blood in cerebrovascular diseases. AB - Ischemic stroke is associated with altered systemic immune responses both early after the onset and in the recovery phase. Interleukin (IL)-10, a Th2 related cytokine, has multiple effects on different cell types, including T and B lymphocytes, monocytes, neutrophils and mast cells. IL-4 is another Th2 cytokine that inhibits the synthesis of pro-inflammatory cytokines by Th1 clones. We used enzyme-linked immunospot assays to detect and enumerate blood mononuclear cells (MNC) secreting IL-10 and IL-4 spontaneously as well as after stimulation with myelin basic protein (MBP), considered to be an autoantigen of possible pathogenic importance in, for example, multiple sclerosis, to evaluate the involvement of anti-inflammatory cytokines in ischemic stroke. All patients with ischemic stroke and cerebral hemorrhage had strongly elevated numbers of IL-10 secreting blood MNC compared with healthy individuals. Numbers of MBP-reactive IL 10 secreting blood MNC were also elevated in a proportion of the patients with stroke and hemorrhage. Levels of IL-4 secreting blood MNC did not differ in ischemic stroke versus healthy individuals. The anti-inflammatory IL-10 could play a pivotal role in ischemic stroke as well as cerebral hemorrhage. PMID- 10362898 TI - Direct costs during the first year after intracerebral hemorrhage. AB - OBJECTIVES: Knowledge of resource use and associated costs of treatment, care and rehabilitation at hospitals and in the health care and social service sectors is limited. This study presents data on the total resource use during the first year after spontaneous intracerebral hemorrhage. METHODS: All patients hospitalized because of stroke at a university hospital in Copenhagen, Denmark, during a 1 year period 1994-1995 were included in a database. The patients were followed until 1 year after the stroke, and data on resource use during and after the hospital stay were collected prospectively. This study investigates a subgroup comprising 90 patients with intracerebral hemorrhage. Resource use is described and costs are calculated. RESULTS: The cost of the hospital stay including acute care and rehabilitation had a mean of 90200 DKK (US$16100). The total cost of health care and social services during the first year had a mean of 123200 DKK (US$22000). Costs decreased significantly with age, but when differences of 30 days case fatality between age groups were considered, the association between costs and age disappeared. CONCLUSIONS: The mean cost of treatment, care and rehabilitation during the first year after intracerebral hemorrhage was 123200 DKK, of which the primary hospital stay constituted 73%. PMID- 10362897 TI - Contrast enhanced pulsed Doppler and colour-coded duplex studies of the cranial vasculature. AB - INTRODUCTION: The aim of this study was to assess the effect of Albunex, a vascular contrast agent based on albumin-coated air microbubbles, on pulsed Doppler and colour-coded duplex sonography of the cranial vasculature. METHODS: Twenty healthy male volunteers received intravenous injections of contrast in single doses ranging from 0.08 to 0.30 ml/kg. Pulsed wave Doppler sonography examination and colour-coded duplex sonography were carried out in the right internal carotid artery (ICA) and middle cerebral artery (MCA) before and after i.v. contrast. The relative intensity increase of the Doppler signal was measured in decibels. RESULTS: Transpulmonary passage of contrast occurred in sufficient amounts to enhance the intensity of the Doppler signal significantly, but the duration of this effect was short. Contrast enhancement also improved visualization of both the ICA and MCA in all subjects. For the transcranial examinations, this resulted in visualization of a greater length of the middle cerebral arteries and additional vessels in the Circle of Willis. CONCLUSIONS: These results confirm that contrast enhancement can significantly improve the quality of Doppler examination and colour-coded duplex sonography of both the intracranial and extracranial vessels. However, the use of Albunex in neurosonology will be of limited value due to its relatively short duration. PMID- 10362899 TI - Magnetic resonance outcome of new enhancing lesions in patients with relapsing remitting multiple sclerosis. AB - The aim of the study was to monitor the natural history of new enhancing lesions in multiple sclerosis (MS) by means of serial gadolinium-enhanced magnetic resonance imaging (MRI). Out of the 63 new enhancing lesions seen on the baseline scan, belonging to 26 relapsing-remitting MS patients, 26 (40%), nine (14%) and four (6%) lesions showed persisting enhancement at first, second and third follow up scan, respectively. At the end of 5 months of follow-up, 58 (92%) of the new enhancing lesions were detected as T2 hyperintensities, 24 (38%) as T1 hypointensities ('black holes'), and five lesions (8%) disappeared in both T2 and T1 weighted images. Duration of gadolinium enhancement of at least two consecutive scans significantly influenced the development of 'black holes'. No significant correlation was observed between volume, location, configuration of enhancement at baseline and final outcome of the lesion. In individual cases, different evolution of new enhancing lesions was observed at the same time. In conclusion, this study documented that different outcomes of new lesions are unrelated either to the individual patient or to the baseline MRI characteristics. However, prolonged blood-brain-barrier disruption as shown by persisting enhancement significantly influences the lesion outcome. PMID- 10362900 TI - Serial SFEMG studies of orbicularis oculi muscle after the first administration of botulinum toxin. AB - Serial single fiber electromyography (SFEMG) examinations of orbicularis oculi muscle in patients with blepharospasm or hemifacial spasm treated with botulinum toxin injections were performed. The aim of the study was to evaluate the impairment of neuromuscular transmission, to follow reinnervation after botulinum toxin administration and to find out whether there was a relationship between SFEMG parameters and clinical symptoms. Examinations were performed before injection, during early and late remission of symptoms, and after recurrence of the involuntary movement. Severe impairment of neuromuscular transmission, as revealed by increased jitter and increased presence of abnormal potential pairs and pairs with blocking, was found in early remission, but fiber density remained unchanged when compared with pretreatment values. In late remission, increased fiber density was registered for the first time. The recurrence of involuntary movements was related to the further increase of fiber density and tendency to normalization of jitter parameters. The study therefore suggests that formation of new neuromuscular junctions and their functional maturation is responsible for muscle recovery after botulinum toxin administration. PMID- 10362901 TI - Glucocorticoid receptor concentrations in muscle biopsies from patients with neuromuscular diseases. AB - Increases in circulating glucocorticoids promote catabolism, particularly in skeletal muscle. The sensitivity of the muscle to glucocorticoids can be altered by a change in the number of glucocorticoid receptors in the muscle, or by a change in the proportions of activated receptors (between binders IB and II). We have investigated the concentration of glucocorticoid receptors, and the proportions of types IB and II, in healthy and diseased muscle. We found significantly reduced concentrations of glucocorticoid receptors in the group of inflammatory myopathies (51% reduction; P < 0.05, Wilcoxon signed rank test). No significant changes in the relative proportions of binders IB and II were found in pathological muscle, although the proportion of binder IB tended towards elevated values (especially in the diabetic neuropathies, with a 17% increase). We conclude that the sensitivity of muscle to glucocorticoids can be reduced in neuromuscular diseases, especially in myositis, by a reduction in the number of glucocorticoid receptors in the tissue, but that no relevant shift in the relation between activated receptor types is present. This could be important in relation to the risk of a secondary steroid myopathy and catabolism of skeletal muscle in the treatment of inflammatory myopathies with glucocorticoids. PMID- 10362902 TI - Psychogenic pseudoepileptic seizures: clinical and electroencephalogram (EEG) video-tape recordings. AB - This paper presents a clinical and electrophysiological analysis of type and duration of seizures recorded by means of long-term video electroencephalogram (EEG) monitoring, a method which enables accurate diagnosis of psychogenic pseudoepileptic seizures occurring with or without epileptic seizures. Analysis is based on 1083 patients, hospitalized at our department between 1990 and 1997, with a preliminary diagnosis of epilepsy. Psychogenic pseudoepileptic seizures were diagnosed in 85 patients (7.8%). In 48 patients, pseudoepileptic seizures alone were diagnosed (group 1), whereas 37 patients had a mixed condition in which pseudoepileptic seizures were accompanied by epileptic seizures (group 2). For comparison of duration of pseudo- and epileptic seizures a control group (group 3), consisting of 55 patients randomly selected from the population of patients suffering from epileptic seizures alone, was parceled out. Long-term video EEG monitoring was performed in 70 patients. In 55 (79%) of these patients 230 seizures (221 pseudoepileptic and nine epileptic) were recorded. In 30 patients (32%), the diagnosis was based on clinical observation of the seizures and on the number of EEG recordings, including activating procedures such as sleep deprivation, photostimulation, hyperventilation and anti-epileptic drug withdrawal. We found that the duration of epileptic seizures was significantly shorter than the duration of psychogenic pseudoepileptic seizures. Our study has exposed the difficulties involved in the diagnosis of psychogenic pseudoepileptic seizures and the negligible value of neuroimaging techniques and interictal EEG recordings in the differential diagnosis of epileptic versus nonepileptic seizures. In this study, psychogenic seizures were significantly more frequent in women than in men; patient history analysis did not confirm the hypothesis that sexual abuse may cause psychogenic seizures. PMID- 10362903 TI - Infarct volume and functional outcome after pre- and postoperative administration of metyrapone, a steroid synthesis inhibitor, in focal brain ischemia in the rat. AB - High blood levels of glucocorticoids are associated with increased mortality, confusion and poor functional outcome in stroke patients. It has been proposed that inhibition of glucocorticoids in acute stroke might be beneficial, but experimental data are conflicting and no long-term follow-up study has been reported. We have studied whether pre- or postoperative administration of metyrapone, a steroid synthesis inhibitor, can influence long-term outcome after ligation of the right middle cerebral artery (MCA) distal to the striatal branches in hypertensive rats. Metyrapone (200 mg/kg) was administered either 30 min before or 1, 12 and 24 h after MCA occlusion. Limb placements and ability to traverse a rotating pole were evaluated pre- and postoperatively. Infarct size, histology and GFAP immunoreactivity were evaluated on 5 microm coronal sections from brains perfused in situ 4 weeks after the ischemic event. Pretreatment did not influence outcome, whereas postoperative administration of metyrapone significantly increased infarct volume (P < 0.05). Post-treated rats performed significantly worse than vehicle-treated rats on the rotating pole 3 weeks after the operation (P < 0.05). Our results do not support the hypothesis that inhibition of glucocorticoid synthesis improves outcome after cerebral ischemia. PMID- 10362904 TI - Quality of life in Alzheimer's disease. PMID- 10362905 TI - CTLA-4 dimorphisms in gammopathy-associated peripheral neuropathy. PMID- 10362906 TI - Serum levels of beta-carotene, alpha-carotene and vitamin A in patients with Alzheimer's disease. AB - To elucidate the possible role of carotenoids and vitamin A as risk factors for Alzheimer's disease (AD), we compared serum levels of beta-carotene and alpha carotene, and vitamin A, measured by isocratic high performance liquid chromatography, of 38 AD patients and 42 controls. The serum levels of alpha carotene did not differ significantly between AD patients and control groups. However, the serum levels of beta-carotene and vitamin A were significantly lower in the AD-patient group. These values did not correlate to age, age at onset or score on the MiniMental State Examination. Weight and body mass index were significantly lower in AD patients than in controls. These results suggest that low serum beta-carotene concentrations in AD patients could be related to a deficiency in dietary intake of this provitamin, although its possible relationship with risk for AD could not be excluded. PMID- 10362907 TI - Familial Guillain-Barre syndrome. AB - There are few reports of Guillain-Barre Syndrome (GBS) occurring in families. We have encountered a mother, who developed acute inflammatory demyelinating polyradiculoneuropathy at age 35 years, whose son developed the bulbar form of GBS 7 years later. Both shared HLA DR2. PMID- 10362908 TI - A patient with multiple sclerosis and Down's syndrome with a rare paroxysmal symptom at onset. AB - Down's syndrome (DS) is often associated with autoimmune diseases, although an association with multiple sclerosis (MS) has not been previously reported. A 49 year-old male with DS experienced progressively worsening gait and bladder dysfunction. Following Poser criteria, the patient was diagnosed with laboratory supported definite MS. Ten days following diagnosis the patient experienced dysestetic paroxysmal pain at the pelvic level (an uncommon complaint in MS) which was initially addressed with carbamazepine, resulting in mild relief and adverse effects consisting of increased motor deficit and decreased daytime alertness. A titration combination of lamotrigine and gabapentin, two relatively new antiepileptic drugs which have been utilized individually for a number of neurological symptoms, resulted in significant reduction in pain frequency and intensity, with no adverse effects. This case study presents details of the first reported association of DS and MS, between which the pathogenetic relationship remains unclear. The presence of a rare symptom complaint in MS, as well as the effective combination of lamotrigine and gabapentin for treating this symptom, without adverse effects is an additional interesting aspect of this case. PMID- 10362909 TI - Generalized freezing in Hallervorden-Spatz syndrome: case report. AB - We report the case of a female patient who was exhibiting slowly progressive, severe, generalized freezing of voluntary movement and disequilibrium. Brain magnetic resonance imaging showed a very low signal intensity in both pallidal nuclei, with a high signal intensity in the central portion, the so called 'eye of-the-tiger' sign. Despite the unusual clinical features, we believe that this is a case of Hallervorden-Spatz syndrome, although without neuropathological examination we were unable to confirm the diagnosis with certainty. This case further demonstrates the complex role of the pallidum in voluntary movement. PMID- 10362910 TI - Primary progressive aphasia: a case report. AB - We report a 69-year-old male patient whose motor aphasia started at the age of 61. The language disability remained isolated and progressed over a period of eight years without any additional cognitive deficits. Computed tomography (CT) and magnetic resonance imaging (MRI) showed moderate cortical atrophy with frontal dominance. Single photon emission tomography (SPECT) showed hypoperfusion in the frontotemporoparietal region, positron emission tomography (PET) demonstrated a global cortical reduction of glucose utilization with a lesser decrement in the occipital lobes. The clinical symptoms and the neuropsychological findings fit the diagnosis of primary progressive aphasia. PMID- 10362911 TI - Transient amnesia triggered by acute marijuana intoxication. AB - We report an unusual case of sudden isolated transient amnesia triggered by acute marijuana use. The memory disorder, apart from the long duration, had the characteristics of a transient global amnesia-like episode. Acute marijuana intoxication can affect memory more globally and severely than previously reported. PMID- 10362912 TI - Issues relating to the assessment of migraine recurrence following triptan therapy. PMID- 10362913 TI - Spontaneous erections and libido increase associated with venlafaxine. PMID- 10362914 TI - We've come a long way, 'ladies': mentoring and nurturing nursing leaders. PMID- 10362915 TI - Crossing state lines: are interstate licenses in nursing's future? PMID- 10362916 TI - Getting into nursing research: dropping in on AWHONN's Continence for Women researchers. PMID- 10362917 TI - Cesarean births: reducing incidence while improving outcomes. PMID- 10362918 TI - How competent are you (or your staff) with shoulder dystocia? AB - Shoulder dystocia-when the fetal head retracts or recoils against the maternal perineum ("turtle sign") and external rotation is not accomplished-occurs in approximately 1 of every 200 deliveries. It's often diagnosed after the emergence of the fetal head when delivery is prevented by impaction of the fetal shoulders within or above the maternal pelvis. When it occurs, shoulder dystocia is an obstetric emergency. PMID- 10362919 TI - Childhood sexual abuse: surveying its impact on primary care. PMID- 10362920 TI - Working with nursing students. Do nurses really eat their young? PMID- 10362921 TI - Surfing the consumer Web. PMID- 10362922 TI - Mentoring: building nursing's future now. PMID- 10362923 TI - Battered nurses: new research shows those giving care may need it most. PMID- 10362924 TI - Health care for elderly people should be free. PMID- 10362925 TI - Do newly qualified nurses have the clinical skills? PMID- 10362926 TI - Health care should be rationed. PMID- 10362927 TI - Health care should not be rationed. PMID- 10362928 TI - Rationalizing the purchase and use of gloves in health care. AB - The last two decades have seen a substantial increase in the use of protective gloves in healthcare establishments. This is attributed to the widespread publicity regarding the risks of transmission of human immunodeficiency virus and hepatitis B and C from blood and body fluid contact. Further, directives from official agencies stressing the need to protect staff from blood-borne viruses in the workplace led to the introduction of a variety of disposable gloves, each claiming total or adequate protective ability. The resulting sensitization to latex in some users has caused concern. This article reports on the process used within one NHS trust to explore glove use and determine the level of latex sensitization among staff. The audit identified an irrational approach to glove purchasing as well as inappropriate use of different types of gloves. The audit was followed by a trial aimed at rationalizing both the purchase and use of gloves. PMID- 10362929 TI - Acute surgical wound care. 4: The importance of documentation. AB - This article, the last in a series of four, discusses the importance of documenting wound care. Studies have shown that nurses do not document wound care as often, or as accurately, as they should in order to comply with the UKCC's (1998) Guidelines for Records and Record Keeping. Although some wound assessment charts have been published and are in use, there is still concern about the validity or reliability of some of these charts. Studies show that further research is necessary in order to validate the charts that are currently in use. An increase in litigation has placed more emphasis on accurate record keeping which shows, in detail, the wound care that is given to each patient. Patients also want to be more informed about their treatment, and this can be done through the use of clinical pathways or multidisciplinary documents. This article also discusses the factors that have to be considered when putting a wound care chart together and gives some examples of existing charts. PMID- 10362931 TI - The patient-centred care model: holistic/multiprofessional/reflective. AB - Holistic patient-centred care (PCC) is often regarded as the quintessence of nursing practice. However, closer examination reveals that provision of care for patients with a physical illness or disability is centred around their compromised physical condition rather than their individual needs. Often, little attention is given to 'total care' and negotiation with the patient is largely excluded. This is evident in the continued use of medical models of care and the assumption that physical needs are the priority for practice. This article presents an alternative--the PCC model. This is a multiprofessional, reflective model which facilitates an unbiased and non-presumptuous approach, thereby overriding the tendency of healthcare professionals to rely on paternalistic and habitual practice. It empowers patients, allowing them to determine their own needs, and encourages reflection, self-awareness and personal and professional growth in the healthcare professionals who adopt it. Although the model was devised and piloted within a hospice it has the potential to be adapted for use in any healthcare setting. PMID- 10362930 TI - Best practice guidelines. AB - Best practice guidelines and multidisciplinary pathways of care are becoming an established and essential feature of clinical practice. They can be seen in a wide variety of clinical settings ranging right across the primary, secondary and tertiary health and social care spectrums. The 1997 White Paper places strong emphasis on quality and consistency of care delivery and gives assurances of performance measurements, integrated care (Wilson, 1996) and clinical governance. It suggests making healthcare delivery against national standards a local responsibility and quality of care the driving force for decision making at every level of the service to ensure excellence for patients no matter where the care is provided. A number of controversial issues surround the use of guidelines. Some argue that they are a fetter on clinical discretion, clinical freedom and can lead to the practice of 'cookbook medicine'. Others maintain that they are an essential aid to providing safe and appropriate medical and nursing care. PMID- 10362932 TI - Intuition in nursing relationships: the result of 'skills' or 'qualities'? AB - Maintaining the scientific orientation of health and nursing care systems has resulted in a lack of focus on nurses' intuitive knowledge base. Consequently, confusion exists as to whether intuition in nursing is the result of acquired skills, or is dependent upon the inherent qualities of the individual. In deconstructing the apparently intuitive process of building rapport, neuro linguistic programming (NLP) can offer insight, suggesting that rapport building depends on learnt skills that have moved into unconscious competence. The examination of nurses' intuitive knowledge, in order to identify underlying skills, needs to be just as rigorous as the acquisition of scientific knowledge. Examining intuitive knowledge can be achieved within a framework of reflective practice to counter the inherent dangers of an increasingly scientific approach operating within a caring profession. PMID- 10362933 TI - Pressure sore formation in the operating theatre: 2. AB - The incidence of pressure sores is seen as a key quality indicator by the Department of Health (1993). The effects of pressure are dependent on its intensity and duration and are widely acknowledged as contributing to pressure sore formation. Therefore, all patients undergoing surgery should be regarded as being at risk. The first article in this two-part series, outlined the pathophysiology of pressure sores and the contributory factors present within the operating theatre (Vol 8(4): 211-17). This article suggests nursing interventions to reduce the incidence of pressure sore formation during the perioperative period. PMID- 10362934 TI - Algosteril calcium alginate dressing for moderate/high exudate. AB - Algosteril is a new alginate dressing manufactured by Les Laboratoires BROTHIER and distributed by Beiersdorf Medical. It is a natural, pure, non-woven dressing made from calcium alginate fibres. It complements other products in the Beiersdorf wound care family such as Cutinova, Cutifilm and Cutisorb. Algosteril rapidly absorbs and retains wound fluid to form an integral gellified structure, thereby maintaining an ideal moist wound healing environment. It traps and immobilizes pathogenic bacteria in the network of gellified fibres, stimulates macrophage activity and activates platelets, resulting in haemostasis and accelerated wound healing. PMID- 10362935 TI - Self-concept and cancer: understanding the nursing role. AB - An experience of cancer can be personally challenging and devastating. Nurses are the ideal health professionals to guide patients through this period in their life because they are in the unique position of being an immediate source of support in patient care. It is therefore essential for nurses to understand the challenge that cancer presents to patients' self-conception in order to facilitate coping and adaptation. This article explores challenges such as investigation and diagnosis of cancer through to treatment, and guides the nurse in aiding the patient in self-conceptual adaptation using Roy's Adaptation Model as a blueprint for care. It also draws upon research evidence and personal views. PMID- 10362936 TI - Nursing is not positively portrayed to children. PMID- 10362937 TI - Before you touch that keyboard.... AB - New technology often gives rise to new health and safety concerns. The Model-T didn't come equipped with a seatbelt; now it is standard equipment. For years, bike helmets were purely optional; now many jurisdictions legislate their use. We wear seatbelts and helmets to prevent injuries, yet we spend hours each day at a potentially hazardous activity--keyboarding--with absolutely no thought to our own health and safety. This is starting to take a toll. PMID- 10362938 TI - Nursing on the 'Net. AB - Nurses must prepare for a vastly different health care delivery system in the next millennium. The new system will be consumer focused, consumer driven and technology enabled. Many changes are already underway, with minimal nursing involvement. One such change is Web-based health care. I envision internet nursing, an integral component within an internet health care environment, as an important next step. PMID- 10362939 TI - The Calgary Conjoint Nursing Program. Part I: Spirit of collaboration. AB - On January 26, 1993, a small but significant gathering in Calgary celebrated the government's approval of the Calgary Conjoint Nursing Program. In attendance were faculty and staff of the nursing education programs of the Foothills Hospital School of Nursing, Mount Royal College and the University of Calgary, as well as senior administrators from each of those sites, practising nurses and a member of Alberta's Legislative Assembly. The group was celebrating the culmination of six years of working together toward the realization of a co-operative, collaborative baccalaureate nursing education program. PMID- 10362941 TI - [Cholera++ epidemic in Kenya]. AB - A cholera epidemic in 1997 followed on the heels of the 1992 epidemic that claimed thousands of victims in Kenya. This time, it emerged in the Migori district near the Tanzanian border, when a woman who had married in Tanzania brought her five-month-old baby to visit her parents. The infant, who contracted serious diarrhea and vomiting, died before the mother could reach a dispensary. During the funeral, a perfect opportunity for the disease to spread, those attending observed the traditional ritual of touching the corpse, and then ate and drank with the next of kin. Many developed symptoms of cholera, and several died in the next few days, even before first aid could be administered. At the Public Health Laboratory in Nairobi, analyses confirmed the presence of the Ogawa strain of Vibrio cholerae. Given the global reputation of Medecins Sans Frontieres [Doctors Without Borders] in the field of cholera, the head of public health for Homa Bay District issued a call for help in August 1997, asking the team to provide preventive solutions and assess the gravity of the situation. Despite intense logistical, technical and health initiatives, the epidemic spread like wildfire. Five months after the initial outbreak, in February 1998, two Canadian nurses working for MSF in Homa Bay hurriedly surveyed the situation in Nyanza Province, which has a population of three million. The author accompanied one of these nurses, Joceline Roy, a Quebecer in her forties, on a tour that lasted more than 15 hours. Roy worked conscientiously, with great precision and energy. This narrative conveys much more than the fatigue and hazards of travel in the developing world; it tells the story of an important, but little publicized, aspect of nursing. PMID- 10362940 TI - Health information telephone service for seniors. AB - The use of the telephone as a medium for providing health information and support specifically to seniors is relatively new. Of the more than 80 Canadian services, apparently only two are dedicated exclusively to older people. Recently, nursing faculty from The University of Western Ontario in London, Ontario, conducted a survey to assess the need for, and probable use of, a free health information telephone service for seniors in the community. PMID- 10362942 TI - Defamation actions. AB - Registered nurses working in today's fast paced, high-tech health care environments are aware of their professional accountability and that, at some point, they may be involved in legal proceedings related to their practice. A civil lawsuit alleging negligence is the predominant concern nurses have when they turn their minds to this issue. PMID- 10362944 TI - Challenges in pediatric home care. AB - One of the significant challenges today in home care for people of any age group, is to keep the level of care realistic and manageable for the whole family. As nurses, we need to be aware of the changes in home care, such as who needs it and what their requirements are, and we should further examine the benefits and limitations of using newer technology in the home. PMID- 10362943 TI - Therapeutic touch in an acute care community hospital. AB - Therapeutic touch now has a 25-year history of practice and research. Based on the idea that within all living things there is a drive toward growth, order and wholeness--and that healing is a natural part of this process--therapeutic touch has been shown to induce relaxation, decrease anxiety and speed healing. It also enables caregivers to embrace their compassion and to touch people with effect. PMID- 10362945 TI - "One who gives comfort to others". Interview by Barbara Sibbald. PMID- 10362947 TI - Cancerlink: helping people help themselves. PMID- 10362946 TI - WHO Programme on Cancer Control: developing a global strategy for cancer, a review. PMID- 10362948 TI - The Dutch Cancer Society. PMID- 10362949 TI - A correlational evaluation of tiredness and lack of energy in survivors of haematological malignancies. AB - Data about tiredness and lack of energy from 91 bone marrow transplant (BMT) survivors and 73 patients receiving maintenance chemotherapy were collected. A correlational evaluation revealed that these two distressing symptoms were associated with physical, psychological, cognitive and social dimensions of quality of life (QOL). A stepwise regression model for BMT survivors showed that both tiredness and lack of energy could be predicted by the combined effect of difficulty concentrating and overall psycho-social adjustment. In addition, dizziness was also influencing tiredness. Lack of energy was predicted in the chemotherapy patients with the combined effects of adjustment to social environment, shortness of breath and psychological symptom distress (R2 = 0.80). In the same group of patients, tiredness was explained by a model consisting mainly of physical symptoms and cognitive symptoms, associated probably with the chemotherapy they were receiving, together with social adjustment (R2 = 0.86). The identification of the reasons behind tiredness and lack of energy in cancer patients, broadly defining fatigue and commonly experienced by them, will have implications for both patient education and the design of appropriate interventions to combat fatigue. PMID- 10362950 TI - Methadone titration in opioid-resistant cancer pain. AB - AIM: To assess the use of methadone in patients with cancer pain who fail to respond to increasing doses of other opioids or experience intolerable side effects from them. METHOD: Inpatients of a specialist palliative care unit were titrated onto oral methadone. The dose was calculated as 10% of the previous morphine equivalent dose, up to maximum of 40 mg, given every 3 h as required for analgesia. When daily requirements were stable it was divided into two regular doses. Pain was assessed on a five-point verbal rating score (VRS): a good response was defined as a fall in VRS of two points or more. Results are expressed as median (range). RESULTS: Thirty-three patients (13 men, 20 women, age 61 (34-91) years), 26 with inadequate analgesia and seven with intolerable opioid related side-effects, were converted to methadone from diamorphine (12), morphine (19) or fentanyl (two). Morphine equivalent dose was 480 (20-1200) mg/day prior to titration. Pain was neuropathic (11), nociceptive (three) or mixed (19). Stabilisation on methadone was complete in 3 (2-18) days in 29 (88%) patients at 80 (20-360) mg/day. Twenty-six (78%) had a good response. Four (12%) patients were withdrawn during titration (three entered terminal phase, one failed to respond). During follow-up 15 (45%) required alteration of methadone dose. Twenty-three (70%) patients were discharged home at 12 (4-26) days. In all cases the stable dose of methadone was less than the previous morphine equivalent, and there was a weak correlation between them. CONCLUSIONS: This method of methadone titration often results in improved pain control in patients with morphine resistance or intolerance. It requires careful titration in a specialist inpatient unit as there is no reliable formula for dose equivalence. PMID- 10362951 TI - Teaching a research-based approach to the management of breathlessness in patients with lung cancer. AB - Breathlessness is a common but complex symptom experienced by patients with lung cancer. Corner and colleagues have developed a therapeutic nursing intervention for the management of breathlessness. The Macmillan Practice Development Unit (MPDU) have recently undertaken a multi-centre evaluation of the intervention. The authors were involved in this multi-centre study as clinical practitioners offering the intervention in a randomised control trial. There is strong evidence to suggest that much research fails to influence clinical practice. A commitment to practice development led us to explore the reasons for this research-practice gap. This paper considers the factors that facilitate implementation of research and, in particular, how this applies to the breathlessness intervention. Practitioners need to feel competent in the skills described in the intervention to implement the research findings into their practice. Therefore, we developed a course to teach the breathlessness intervention. A description of the course is presented, together with the theoretical basis for the course design and the tools used to evaluate it. PMID- 10362953 TI - The Palliative Care Education Group for Gloucestershire (PEGG): an integrated model of multidisciplinary education in palliative care. AB - Multidisciplinary teamwork is an essential part of palliative care. Traditionally, in the UK, education of health care professionals has been carried out within single professional disciplines. The Palliative Care Education Group (PEGG) for Gloucestershire, a county in the south-west region of England, UK, is a multidisciplinary group, which promotes and integrates palliative care education across the county. This report describes the evolution of the group, its aims and objectives and the methods used to evaluate its impact on clinical practice. PEGG is a practical model of multidisciplinary education, which could be adopted in other areas. PMID- 10362952 TI - The experience of breathlessness in lung cancer. AB - Breathlessness is a common problem in advanced cancer ranked amongst the 10 most common symptoms in patients admitted to palliative care units. Alongside coughing, it is the most commonly reported symptom in lung cancer. Despite the prevalence of breathlessness, little research has been undertaken on the experience of the symptom, or on the restrictions it imposes on daily life. The data reported in this paper were collected as part of a study piloting new non pharmacological intervention for patients with breathlessness as a result of lung cancer. Data on the experience of breathlessness from assessment notes recorded by nurses during conversations with 52 patients with lung cancer, were analysed using content analysis. Both physical and emotional sensations were associated with descriptions of breathlessness, such as the feeling of being unable to get enough breath, or of panic or impending death. Breathlessness was only continuous in eight patients, the remainder (85%) had an intermittent pattern of the symptom, usually triggered by exertion or, less commonly, emotion. Numerous restrictions on activity were reported as a result of breathlessness, on functioning inside and outside the home, to social life, and its implications for feelings about oneself. Most patients had attempted to find ways of managing the problems for themselves since, prior to receiving nursing intervention, little or no help had been forthcoming from health professionals. PMID- 10362954 TI - Continuing medical education by satellite: implications for oncology education. AB - The need for rapid dissemination of medical information is linked with the requirement for training and career grade doctors to be up-to-date with their knowledge. Over 220 sites in Europe receive EuroTransMed oncology programmes, during the academic terms. The non-profit foundation brings together expert panels, providing the latest evidence for best practice. Viewers are able to interact during the programme with the panel members using phone, fax, ISDN and e mail. Increasing use of telematics has helped in the development of this important quality learning initiative for doctors in Europe, and addressed the continuing education need of oncology health professionals. PMID- 10362955 TI - Forum for Applied Cancer Education and Training. Critical appraisal. PMID- 10362956 TI - Another 10 tips to think more critically. PMID- 10362957 TI - HHNA. Psychiatric home care nursing position statement. PMID- 10362958 TI - New 1999-2000 standard: protecting the integrity of clinical decision-making. PMID- 10362959 TI - Welcome to OASIS. Dos and don'ts. PMID- 10362960 TI - Congress's report card on home care progress. PMID- 10362961 TI - Identifying and evaluating wound infection. AB - Wound infection is a serious problem in the home care patient. Early detection of infection, prompt treatment, and continuous surveillance are pivotal to elimination of the infection and prevention of complications. This article addresses identification of wound infection in postsurgical and chronic wounds based on data obtained from the patient's history, physical examination, and diagnostic tests. Appreciating the overt and subtle signs and symptoms of wound infection and acting on them is critical to timely diagnosis and treatment. PMID- 10362962 TI - Remediating the effects of sarcopenia in the elderly client's plan of care. AB - Because home care nurses are in close contact with frail elders, it is important that they recognize sarcopenia and understand options available to remediate the dangerous consequences of this condition. This article describes an overview of human aging and muscle mass loss, the effects of regular exercise on muscle mass, muscle strength, and functional capacity, and implications for creating interdisciplinary plans of care that impact outcomes. PMID- 10362963 TI - Stories at the hour of our death. AB - Storytelling through reminiscence and life review can be an important activity for individuals who are dying. Benefits include catharsis, integration, meaning making, and the enhancement of relationships. Specific tools for facilitating life stories--taped autobiographies, journals, "reminiscentia," and scrapbooks- are discussed. PMID- 10362964 TI - The Neal theory: implications for practice and administration. AB - New research demonstrates that home health nurses proceed through a three-stage process to become successful and autonomous in their practice. It also indicates that they must be adaptable in order to function effectively and successfully in home care. This research has many implications for clinicians, managers, and administrators. PMID- 10362965 TI - The oldest profession. PMID- 10362966 TI - Documentation tips. PMID- 10362967 TI - Listen to the messages within your agency. PMID- 10362968 TI - Celebrating nursing research. PMID- 10362969 TI - Katz Index of Independence in Activities of Daily Living. PMID- 10362970 TI - Caregiver distress. Related to disruptive behaviors on special care units versus traditional long-term care units. AB - The link between staff stress and exposure to disruptive behaviors is an important issue in long-term care settings. This study compared the perceptions of two groups of formal caregivers (staff) regarding their distress from the behaviors of residents in their care. Staff on special care units for dementia were less distressed with disruptive behaviors than comparable staff on traditional units, although they reported higher exposure to these behaviors. These results were related to different perceptions of intent to harm and expectations of physical aggression as "part of the job." Implications for nursing include education and support for staff to enhance the quality of life for residents and staff on units where disruptive behaviors occur. PMID- 10362971 TI - Nursing interventions for depression in newly admitted nursing home residents. PMID- 10362972 TI - Developing individualized care in nursing homes: integrating the views of nurses and certified nurse aides. AB - Despite recent attention devoted to the development of individualized care in nursing homes, empirical research assessing changes in practice is quite limited, and very few studies have explored specifically the experiences and perceptions of certified nurse aides (CNAs). This study reports findings from a comparative analysis conducted on a data set including quantitative and qualitative data from CNAs (N = 289) and nurses in Connecticut (N = 245). Measures of obstacles to individualized care and needs for future supports were explored. A number of significant differences in perceptions of obstacles to providing individualized care were found. The nurses were significantly more likely to identify the following impediments to change: cost (p < .0001), concepts not integrated into work (p < .0001), lack of administrative support (p < .10), and staff attitudes (p < .10). The CNAs were significantly more likely to report inadequate staffing (p < .001), lack of interdisciplinary teams (p < .001), and resident and family attitudes (p < .01) as problematic. These findings suggest substantial discordance among nurses and CNAs on a number of important issues surrounding individualized care. Such disparate perceptions pose challenges to nursing homes committed to the implementation of individualized care alternatives. Successful approaches must consider the various vantage points of caregivers and administrators. PMID- 10362973 TI - Bladder diaries and voiding patterns in older adults. AB - Urinary incontinence is one chronic health problem that many older adults experience. Assessment is the first step in approaching a urinary elimination problem and forms the basis for determining potential management modalities. The bladder diary, one type of fluid intake and output record, is helpful in describing an individual's voiding pattern and is an integral component of urinary assessment. Bladder diaries also can be used to evaluate the effectiveness of interventions initiated to manage incontinence. This article describes the procedure for using bladder diaries within the context of a research study conducted with residents of retirement settings. PMID- 10362974 TI - Evaluation and management of orthostatic hypotension in elderly individuals. PMID- 10362975 TI - How do you attempt to bring some normalization into your residents' lives (e.g., promotion of the familiar, allowing some control in decision-making)? PMID- 10362977 TI - Licensed practical/vocational (male) nurse ... umm ... I like the sound of that! PMID- 10362976 TI - Obtaining a positive relationship with your supervisor. AB - Obtaining a satisfactory relationship with the supervisor is essential for the LP/VN who has just taken a job. Both personal and job satisfaction will result when a compatible trusting relationship can be obtained. How to effectively disagree with a supervisor is a skill every employee should have. It is especially important for the LP/VN who may feel the supervisor's idea may adversely affect patient care, agency function, or personal effectiveness in performance. Strategies have been described for obtaining a positive working relationship with the supervisor. PMID- 10362978 TI - Benevolent power. AB - The nurse's ability to use "benevolent power" can be a very useful tool in relating to patients. Use of "benevolent power" takes advantage of the nurse's power and authority by virtue of positions However, rather than focusing on the nurse's needs or manipulating the patient in a negative way, use of "benevolent power" allows the nurse to respond to the patient situation in a way which is beneficial and positive for the patient. PMID- 10362979 TI - Freedom from the chain of septic flow: handwashing in infection control. PMID- 10362980 TI - Ischemic venous thrombosis: its hidden agenda. AB - Ischemic venous thrombosis (IVT) is the most severe form of venous thrombosis. The purpose of this article is to describe this rare clinical condition and to review 14 cases of IVT that involve the lower extremities, emphasizing its association with cancer. Between September 1995 and November 1997, 9 women and 5 men, ranging in age from 37 to 75 years (mean 58.6 years), were treated for IVT. Malignancy was the most prevalent risk factor, occurring in 86% (12 of 14) of the patients, and was not diagnosed in 58% (7 of 12) of the cases until after IVT occurred. A postoperative state and a previous history of thrombophlebitis were the next most common risk factors and were equal in frequency (39%). Venous gangrene was present in 79% (11 of 14) of the cases, resulting in 11 amputations in 9 patients. The mortality rate was 71% (10 of 14) of the patients, with 40% of the deaths occurring within 2 months of the diagnosis of IVT. These data support the belief that a strong association exists between IVT and cancer and that IVT may precede the diagnosis of cancer, representing a potential marker for an undiagnosed malignancy. Furthermore, the combination of extensive venous gangrene and disseminated cancer has a poor prognosis. PMID- 10362981 TI - Venous ulcers: etiology and care of patients treated with human skin equivalent grafts. AB - Care for a patient with a venous ulcer is complex, necessitating collaboration of a multidisciplinary team to achieve the goal of providing comprehensive wound care and optimally managing complications, current conditions, and healing time. Patients often have venous ulcers for a long time, and they frequently have multiple diagnoses. Chronic ulcers that do not heal necessitate closure with a graft. Apligraf, (Novartis Pharmaceutical Corp, East Hanover, NJ), a human skin equivalent, is often used in nonhealing or difficult-to-heal ulcers. Knowledge of how to care for the grafted wound and protection of the grafted site is essential. PMID- 10362982 TI - Tissue perfusion as a key underlying concept of pressure ulcer development and treatment. AB - The purpose of this article is to refine and advance the theory that tissue perfusion is the key concept in the development and delayed healing of pressure ulcers. The person likely to have (be at risk for) pressure ulcers is at greater risk for inadequate tissue perfusion generally and specifically at pressure points. Accordingly, the tissue perfusion theory of pressure ulcer development states that the factors that contribute to inadequate tissue perfusion should be used to predict (identify risk factors for) pressure ulcer development and delayed healing. Factors influencing a person's adequacy of tissue perfusion need to be assessed to identify risk for pressure ulcers. In addition, adequate tissue perfusion needs to be maintained to provide for healing of such wounds. Current beliefs about the causes and prevention of pressure ulcers are described. Physiologic components of the tissue perfusion theory are discussed: cellular exchange of nutrients and wastes, autoregulation of blood flow at the cellular level, and regulatory mechanisms that affect tissue perfusion when it is significantly compromised. The North American Nursing Diagnosis Association (NANDA) framework is used to classify or group examples of common pathophysiologic, treatment-related, situational, and maturational factors. Implications for research, practice, and education also are discussed. PMID- 10362983 TI - Effect of povidone-iodine on wound healing: a review. AB - For the purpose of providing a summary of current clinical trials to determine whether povidone-iodine is beneficial or detrimental to wound healing, an integrated review was completed. Clinical trials were defined as any study that uses some concentration and form of povidone-iodine in a comparison or evaluation with other products or treatments resulting in an impact of povidone-iodine on wounds. The use of povidone-iodine for cleansing, irrigating, and dressing wounds is controversial. Wound healing is complex and requires safe and effective treatment modalities. Numerous in vitro and in vivo studies have been done with conflicting results on bactericidal effects and cytotoxicity of this antimicrobial agent. Human and animal in vivo studies in the last 10 years were used for this review because often the relevance of in vitro data in clinical conditions are questioned. The varied studies provide evidence that in most instances, povidone-iodine did not effectively promote good wound healing; in fact, most studies showed either impaired wound healing, reduced wound strength, or infection. PMID- 10362984 TI - Assessing practice in preventing pressure damage. PMID- 10362985 TI - Pressure sore survey. Part 3: Locus of control. AB - This is the third in a three-part article which investigates the prevalence, knowledge and attitudes to pressure sores in one NHS trust. This study describes the methodology used in choosing and developing attitude scales to explore whether there are any relationships between the locus of control and pressure sore prevention. Factors to do with attitude and the value associated with pressure sore prevention have a central role. Attitudes and beliefs affect what we do and may contribute to pressure sore development. PMID- 10362986 TI - Haemorrhagic cellulitis. PMID- 10362988 TI - Managing MRSA. PMID- 10362987 TI - A systematic review of laser therapy for venous leg ulcers. AB - A systematic review of randomised controlled trials (RCTs) was conducted to establish the effectiveness of low-level laser therapy as a treatment for venous leg ulcers. Wound-care journals, conference proceedings and electronic databases (including Medline and Cinahl) were searched up to October 1997 for RCTs comparing low-level laser therapy with sham laser, no laser, or non-coherent light of other wavelengths. In addition, companies who manufacture or distribute therapeutic lasers were contacted for any unpublished or ongoing studies. Results from searches were scrutinised by one reviewer to identify possible RCTs and full reports of these were obtained. Details of eligible studies were extracted and summarised using a data extraction sheet. Data extraction was checked by a second reviewer. Meta-analysis was used to combine the results of trials where the interventions and outcome measures were sufficiently similar. A total of four eligible RCTs were identified. Two compared treatment with laser therapy to sham or placebo laser treatment. One study compared laser therapy with ultraviolet therapy. The fourth was a three-armed study which compared the effects of laser therapy alone, laser therapy plus infrared light, and non-coherent unpolarised red light. The comparisons of laser therapy with placebo, and laser therapy with ultraviolet therapy, showed no significant difference between treatments with regard to ulcer healing rates. The comparison of laser with red light showed a significant increase in complete healing at nine months for the combination of laser and infrared light compared to non-coherent unpolarised red light. We have not found any evidence of the benefit of low-level laser therapy per se on venous leg ulcer healing. It appears that a combination of HeNe laser and infrared light may promote the healing of venous ulcers, however more research is needed. PMID- 10362989 TI - A seating assessment tool for community use. PMID- 10362990 TI - Assessing the seated patient for the risk of pressure damage. PMID- 10362991 TI - Pressure sore prevention in hospital patients: a clinical audit. AB - Pressure sores cause significant mortality and morbidity as well as being a financial burden on health-care services. Reduction of pressure sore incidence is a Department of Health priority. Pressure sores are accepted as largely preventable complications of illness and disability and the means to achieve prevention are available. The aim of this clinical audit was to identify potential contributing factors to pressure sore acquisition in an acute hospital setting. The results suggest that substantial changes in the approach to clinical management may be needed. PMID- 10362992 TI - Graduated compression and the prevention of deep vein thrombosis (Part 3). PMID- 10362993 TI - Enterocutaneous fistulae. PMID- 10362994 TI - Delayed primary closure in the management of gunshot wounds. PMID- 10362995 TI - An introduction to hemoglobin physiology. AB - Hemoglobin plays an important physiologic role, one that begins early in fetal development. The focus of this article is hemoglobin physiology. Developmental erythropoiesis, developmental stages of hemoglobin, and postnatal erythropoiesis and hemoglobin production are discussed. The function of hemoglobin, its affinity for oxygen, and the clinical significance of the oxyhemoglobin dissociation curve are also explored. PMID- 10362997 TI - Assessment of the newborn with dysmorphic features. AB - Accurate assessment of all newborns is essential for identifying those who will need more thorough examination, medical or support services, or family counseling. External assessment of dysmorphic features offers clues to the presence of internal anomalies. This article describes a systematic approach to the assessment of dysmorphic features in the newborn. PMID- 10362996 TI - Exstrophy of the bladder. AB - Exstrophy of the bladder is a rare congenital defect that occurs when the abdominal wall and underlying structures, including the ventral wall of the bladder, fail to fuse in utero. As a result, the lower urinary tract is exposed, and the everted bladder appears through the abdominal opening. Various surgical interventions have been employed with variable success in the hope of achieving complete dryness, full control over delivery of urine, freedom from catheters and external appliances, and a protected upper urinary tract. The most popular surgical approach is the primary bladder closure with secondary bladder neck reconstruction. Comprehensive nursing, medical, and surgical care are necessary to preserve renal and sexual function. The many complex problems experienced by these infants and their families call for a multidisciplinary approach. This article reviews occurrence, clinical presentation, and management of exstrophy of the bladder. PMID- 10362998 TI - Retinopathy of prematurity in the 1990s. AB - Infants diagnosed with retinopathy of prematurity (ROP) are at risk for developing visual impairments. The International Classification of ROP (ICROP) developed the most useful tool available for communicating the location and extent of the condition, with three zones to describe the location and five stages to describe the extent. Despite current knowledge and treatment, however, 500 infants are blinded each year by ROP. Infants at highest risk are the smallest and most immature. The nurse must understand why and how ROP occurs and the forms of management used to treat ROP. This article reports on the incidence, pathophysiology, risk factors, screening, treatment, and long-term complications of ROP in the 1990s. PMID- 10363000 TI - Perinatal substance abuse: working with neonates and families. PMID- 10362999 TI - Camptomelic dysplasia: a case study. AB - Camptomelic dysplasia (CMD) is a congenital short-limb skeletal dysplasia characterized by prenatal bowing of the lower limbs in association with additional anomalies of the tracheobronchial tree or genitourinary tract. Perinatal and early neonatal death from respiratory failure is common. Diagnosis and long-term management of the infant with CMD require a coordinated effort among many specialists. This article presents a general overview of skeletal dysplasias and camptomelic dysplasia. It concludes with a case study illustrating the many problems infants with CMD may have and the complex treatment and follow up services they require. PMID- 10363001 TI - Prospects for PD in the future. PMID- 10363002 TI - PD nursing: what lies ahead for the next millenium? PMID- 10363003 TI - Using subjective global assessment to identify malnutrition in the ESRD patient. PMID- 10363004 TI - Renal community looking to take control of vascular access. PMID- 10363005 TI - From the facility to the network: DxBase moves on to SIMS and VISION to help track patient outcomes. PMID- 10363006 TI - Targeted intervention to improve compliance in an urban hemodialysis population. AB - Noncompliance impacts the successful delivery of care to the "urban" ESRD population. It is our belief that noncompliance can be an extension of the dysfunctional coping mechanisms of patients who lack adequate psychosocial support. In this article, we have described a pilot program providing on-site, targeted intervention for patients identified at increased risk for poor clinical outcomes, based on failure to comply with the prescribed dialysis treatment prescription. This collaboration between our "nephrology practice," an outpatient "HD facility," and a "C-L psychiatry service" has resulted in extremely low no show rates in an urban dialysis population. We have also discovered an opportunity for reducing hospitalizations. Targeted intervention can improve compliance and prove to be cost effective. PMID- 10363007 TI - Legislative challenges facing transplantation in the year ahead. PMID- 10363008 TI - Consider exchanging organ donation for prison time. PMID- 10363009 TI - Why Medicare should cover the kidney-pancreas transplant. PMID- 10363010 TI - A homecoming for hemodialysis? PMID- 10363012 TI - The future for nurse executives. PMID- 10363011 TI - The future of nurse executives. PMID- 10363013 TI - Advancing science forces change for everyone in the health care system. AB - This article discusses expected technological developments in the nursing field. It also provides sources for keeping aware of the developments in science. PMID- 10363014 TI - What is the future for nurse executives? AB - This article describes the future role of nurse executives as designers of health care rather than respondents to internal and external environmental forces. The successful executives will be proactive, dedicated, and politically astute. These individuals will engineer vertical organizations that are empowered, fluid, and flexible. These new organizational skills will be developed through innovative executive educational programs designed at the master's and doctoral levels. Well established executives from business, education, public health, medicine, administration, health systems, and education will become the faculty preparing these individuals. These programs will integrate other health care disciplines through an integrated and flexible approach to learning. The development of these roles will influence policy and decision making at all levels within health care. PMID- 10363015 TI - An innovative professional nursing practice model. AB - Nursing, as with health care delivery, is changing to meet the greater challenges brought about by managed care and the accompanying external forces in the marketplace. Nurses, with their vast resources of knowledge and experience, are crucial in achieving optimal quality care. Given the opportunity to redefine and strengthen the impact of nursing practice at Piedmont Medical Center in Atlanta, Georgia, a group of advanced practice nurses began developing a unique practice model. The resulting Professional Nursing Practice Model--a theoretical framework created for nurses by nurses--presents a common vision of human beings, health, and nursing in accordance with the values and beliefs of nursing. This model demonstrates the importance of a person-centered, value-driven nursing practice across the continuum of health from birth to death. PMID- 10363016 TI - The role of the chief nurse executive in the knowledge-intense organization of the future. AB - The health care world is being upended. Nurse executives, like other leaders, must develop new roles or risk being displaced or discarded. The health care organizations of the future are likely to look and function much more like "knowledge-intense organizations," peopled by knowledge professionals working in sophisticated interdisciplinary groups or teams. Nurse executives are well positioned to create new leadership roles in these knowledge-intense organizations. PMID- 10363017 TI - Health care managers' and administrators' roles, functions, and responsibilities. AB - Health care managers and administrators are increasingly assuming leadership and management responsibilities for multiple units and departments. To determine key roles, functions, and responsibilities of nursing and other health care managers and administrators, a survey was conducted to identify the perceived importance and amount of time these individuals devote to selected activities. Although the amount of time and perceived importance of selected activities varied with each environmental context, some key roles, functions, and responsibilities across practice disciplines did emerge. Regardless of the environmental context, administrators and managers at all levels in health care organizations spend significant amounts of time and place high value on communication, problem solving and decision making, collaboration with other disciplines, people development, and cost containment. PMID- 10363018 TI - How other members of the top management team see the nurse executive. AB - This study compared how top-level organizational groups within Veterans Affairs Medical Center (VAMC) top management teams perceived nurse executives' participation in strategic decision making. Nurse executives' perceptions of their own involvement in strategic decision making generally agreed with the views of both chiefs of staff and directors. However, the associate director's perceptions of the nurse executive's involvement often differed significantly with those of the nurse. Nurse executives will be better prepared to function effectively within the top management team if they are aware of patterns in how they are perceived by other members. PMID- 10363019 TI - Effectiveness of nurse executives: measurement of role factors and attitudes. AB - This article describes the outcome of a survey of 40 nurse executives and 56 influential colleagues. Both groups agreed that leadership was the most important quality for the executive role. The nurses' primary focus was resolution of patient care problems. Influential colleagues stated resource allocation and initiation of change were two qualities needing improvement. The nurses rated themselves high in confidence about their job responsibilities, scored relationships as the most satisfying attribute, and cited provision of quality health care as the greatest advantage of their position. The greatest disadvantage was lack of administrative support. The results suggest the educational preparation needed for nurse executives. PMID- 10363020 TI - Career with no return: roles, demands, and challenges as perceived by Swedish ward managers. AB - The article describes the role of ward managers at middle-sized hospitals and analyzes that role with respect to motivation, role content, and skills and competencies, as well as challenges and problems. The most striking result of this study was the unwillingness of Swedish executive nurses to return to nursing. Among the findings that explain this "career with no return" are an increased taste for challenges; an irrevocable, new professional identity; and a loss of nursing skills and the ability to subordinate. The article discusses the implications for recruitment and education. PMID- 10363021 TI - A multihospital study of length of stay among acute myocardial infarction patients. PMID- 10363022 TI - The joint commission. The future of the nurse executive: a standards focus. AB - A retrospective of joint Commission changes from it's early days to the present is provided in this column. How the Joint Commission on the Accreditation of Healthcare Organizations evolved from the American College of Surgeons initiative to standardize hospital systems and equipment is discussed. Structural standards developed into important functions that crossed departments within an organization. Discussion of the current initiatives and future challenges to improve care to the public is included. PMID- 10363023 TI - Molecular approaches to achieving control of gene expression by drug intervention at the transcriptional level. AB - In this article we first very briefly review current approaches to the design of drugs that have specificity for the modulation of gene expression and selectivity for target cells at the transcription level by targeting DNA. We focus this review on our approaches to gaining selectivity by drug-induced architectural alteration in DNA structure, selectivity achieved by protein-induced changes in DNA structure or dynamics, and hijacking of nuclear receptors. PMID- 10363024 TI - The role of base excision repair in the repair of DNA adducts formed by a series of nitrogen mustard-containing analogues of distamycin of increasing binding site size. AB - The role of base excision repair in the repair of alkylation damage produced by a series of sequence specific oligopyrrole-containing analogues of distamycin A that tether benzoic acid mustard (BAM) has been examined. Whereas BAM alkylates and cross-links in the major groove of DNA, attachment to pyrrole units produces monoalkylations in the minor groove of DNA at AT tracts. Both sequence specificity of alkylation and cytotoxicity increase from one to three attached pyrrole units (compounds 1-3), and with 3 alkylation is selective for purine-N3 in the sequence 5'-TTTTGPu (where Pu = guanine or adenine). In a model bacterial (Escherichia coli) system repair of the sequence specific minor groove alkylations produced by 2 and 3 does not appear to involve BER, since neither a formamidopyrimidine-DNA glycosylase repair deficient E. coli mutant (BH 20, fpg- mutant) nor a 3-methyladenine-DNA glycosylase repair deficient mutant (GC 4803, tag-alkA- mutant) showed increased cytotoxicity to 2 or 3 compared with the wild type, AB 1157. The monopyrrole compound 1 was, however, approximately 4-fold more cytotoxic to the GC 4803 mutant compared with wild type and BH 20, suggesting a role for the 3-methyladenine-DNA glycosylase in the recognition and excision of the adducts formed by 1. In contrast, increased sensitivity (> 10-fold) was observed for the conventional nitrogen mustard BAM in the BH 20 strain, suggesting a role for the formamidopyrimidine-DNA glycosylase in the repair of the lesions produced by the agent. In a cell-free system the E. coli 3 methyladenine-DNA glycosylase (AlkA) was shown to remove alkylations at 5' TTTTGPu sequences. However, the efficiency in removing the adducts formed by the oligopyrrole compounds decreased dramatically from compound 1 to compound 3. Increasing the size of the DNA adduct formed in the minor groove therefore decreased the efficiency of recognition and removal of the adduct by the DNA glycosylase. PMID- 10363025 TI - Bis-benzimidazole anticancer agents: targeting human tumour helicases. AB - Certain DNA minor groove binding agents, distamycin, netropsin, and a series of anticancer bis-benzimidazoles can block DNA helicase activity by binding to duplex DNA at specific base sequences. DNA helicases are crucial to cell DNA replication, transcription and repair because these enzymes separate double stranded DNA, thereby preparing the strands for enzymatic manipulation. From our studies we have developed a hypothesis that focuses on cellular DNA helicase action as a mechanistic site where these minor groove binders can act. A crucial aspect for modulation of DNA activity by drugs is for specificity and selectivity. A series of DNA-interactive bis-benzimidazole analogues of Hoechst 33258 was also prepared to explore the potential for anticancer activity mediated for certain of the drugs via bioreductive activation by endogenous NADH or NADPH. The biological endpoints examined included intracellular distribution in euoxic and hypoxic conditions observed by fluorescence microscopy; relative efficacy as antimetabolites determined by the MTT [tetrazolium salt, 3-(4,5-dimethylthiazol-2 yl)-2,5-diphenyl tetrazolium bromide] assay in euoxic and hypoxic conditions; and relative inhibitory activities on human DNA helicase, as determined by degree of dissociation of GC B6486 DNA. The intracellular distribution was unique to each of the test compounds. Compounds V-93 and V-153, the respective semiquinone and quinone derivatives, demonstrated the predicted enhanced cytotoxicity and anti helicase activities, supporting the concept that preferential binding of DNA at 5'-CG and TG sequences provides a novel approach to anticancer drug development. PMID- 10363026 TI - 5,7-Disubstituted analogues of the mixed topoisomerase I/II poison N-[2 (dimethylamino)ethyl]acridine-4-carboxamide (DACA): DNA binding and patterns of cytotoxicity. AB - DACA is a DNA-intercalating agent and dual topoisomerase (topo) I/II inhibitor currently in clinical trial as an anticancer drug. Substitutions in the acridine ring of DACA have significant effects on biological activity, with 5-substituted analogues being more potent but relatively less active against cell lines that underexpress topo II, and the converse for 7-substituted analogues. A small series of 5,7-disubstituted analogues was therefore prepared and evaluated. The compounds were prepared by CDI-assisted coupling of the appropriate acridine acids. When these contained no or only one halogen atom, they could be prepared by Al/Hg amalgam reduction of the corresponding acridine acids. However, this method could not be used to prepare dihalogen-substituted acridine acids due to substantial dehalogenation, and these intermediates were synthesized via cyclization of the appropriate aldehydes to give the acridines directly. These compounds showed enhanced DNA binding compared with the parent DACA, indicating that the known favourable influence of 5-substituents on DNA binding is retained. Cell line studies showed that the 5,7-disubstituted compounds retained both the broad-spectrum effectiveness of the 7-monosubstituted analogues and the higher cytotoxic potency of the 5-monosubstituted analogues. The 7-chloro-5-methyl and 5 chloro-7-methyl analogues showed comparable in vivo antitumour activity to DACA in the subcutaneous colon 38 model, but were substantially more potent (optimal doses of 60 mg/kg compared with 200 mg/kg for DACA). PMID- 10363027 TI - Relationships between topoisomerase II inhibition, sequence-specificity and DNA binding mode of dicationic diphenylfuran derivatives. AB - Four diphenylfuran derivatives possessing different dicationic terminal side chains were used to investigate sequence-specific binding to DNA and poisoning of human topoisomerase II. Footprinting experiments with a range of DNA substrates attest that all four drugs bind selectively to AT-rich sequences in DNA. However, the quantitative analysis of the footprinting profiles reveals significant differences in terms of AT-selectivity according to the nature of the basic side chains. Furimidazoline (DB60) shows a reduced capacity to interact selectively with A.T tetrads compared with furamidine (DB75) and the 3-pentyl-substituted diamidine analogue DB226. DB244, for which the two amidine ends are substituted with a cyclopentyl group, exhibits the most pronounced AT specificity. It binds tightly to sites composed of at least four adjacent AT base pairs, such as 5' TAAT, AATT and TTTT. At low concentrations (< 2 microM) DB60 is also capable of forming stable complexes with AT sites but at higher concentrations the binding becomes totally non-specific due to additional intercalation of drug molecules into GC-rich sequences. Nevertheless, DB60 is the only drug is the series which stabilizes DNA-topoisomerase II covalent complexes. This compound effectively promotes DNA cleavage by topoisomerase II whereas DB75, DB226 and DB244 have practically no effect. The topoisomerase II poisoning activity of DB60 correlates with its ability to intercalate into GC sites in DNA whereas the three other diphenylfurans essentially behave as typical AT-selective minor groove binders. The study suggests that the antimicrobial activity of the diphenylfurans, which are active against the Pneumocystis carinii pathogen (PCP), depends essentially on their capacity to recognize AT-rich DNA sequences rather than their ability to interfere with topoisomerase II. In contrast, the cytotoxicity of drugs like DB60 would be connected with the formation of intercalation complexes and the stimulation of DNA cleavage by human topoisomerase II. PMID- 10363028 TI - Polybenzamide mustards: structure-activity relationships for DNA sequence specific alkylation. AB - A series of cytotoxic polybenzamide mustards targeted to the minor groove of DNA were used to define structure-activity relationships for sequence-specific DNA alkylation. Compounds with an annular structure closely matched to the minor groove of DNA, and with concave-facing, potentially H-bonding NH groups, had a strong preference for alkylating adenines in sequences possessing four or more consecutive adenines. Two compounds whose annular structure matched that of the minor groove better when at least one carboxamide NH group faced outwards showed a high specificity for the consensus sequence (A/T)A(G/C) (A/T)N. Several compounds also alkylated specific guanines, presumably at the N3 position. Modelling studies suggest the most important contribution to sequence-specific alkylation is the H-bonds formed between these compounds and DNA, with factors such as the degree and positioning of cationic charge being less influential. PMID- 10363029 TI - PNU 157977: a new potent antitumour agent exhibiting low in vivo toxicity in mice injected with L1210 leukaemia cells. AB - The design, synthesis, in vitro and in vivo activity against L1210 murine leukaemia of the dibromo nitrogen mustard derivative of 2, called PNU 157977, is described and the structure-activity relationship discussed. This dibromo derivative is almost two orders of magnitude more cytotoxic than the dichloro counterpart having the same oligopeptidic chain (IC50 2.7 ng/ml versus 225 ng/ml), and it showed in vivo an increased survival time which is 5- and 3-fold longer than that of tallimustine and 2 (and T/C 750 versus 133 and 213) respectively. Moreover PNU 157977 shows activity against the M5076 solid tumour markedly inferior to that of the closely analogous 2. Footprinting experiments conducted using the oestrogen receptor PCR probe as the footprinting target molecule show that PNU 157977 possesses a different sequence-specific alkylation and greater cleavage activity than either 2 or tallimustine. PMID- 10363030 TI - Comparison of the patterns of DNA alkylation by phenol and amino seco-CBI-TMI compounds: use of a PCR method for the facile preparation of single end-labelled double-stranded DNA. AB - Both 5-hydroxy- and 5-amino-seco-CBI-TMI minor groove alkylators are very potent cytotoxins. The patterns of alkylation of the two enantiomers of both compounds were compared on a section of the gpt gene. All of the compounds alkylated only at adenines, with the amino compounds being slightly more selective. Consensus alkylation sequences for both S (natural) enantiomers were identical, but for the R (unnatural) enantiomers these varied slightly. The consensus sequences suggest that the S enantiomers bind lying in the 3'-->5' direction from the alkylated adenine, but there was no clear indication of which direction the R enantiomers lie on the DNA. Both S enantiomers were 10- to 100-fold more efficient alkylators than the R enantiomers, and the amino compounds were somewhat more efficient than the corresponding phenols. The S enantiomers were more cytotoxic then the R in both the phenol and amino series. The large amounts of end-labelled DNA required for this work was obtained by first end-labelling appropriate primer oligonucleotides, then amplifying by PCR. Compared with other methods in use, this is a simple and flexible one-step procedure for the preparation of labelled DNA of any sequence. An improvement in the synthesis of 5-hydroxy-seco-CBI-TMI is reported. PMID- 10363031 TI - The plasminogen activation system in lung cancer--with special reference to the prognostic role in "non-small cell lung cancer". PMID- 10363032 TI - Advances in liver transplantation: overview and status. PMID- 10363033 TI - Advances in kidney transplantation: renal failure in the United States. PMID- 10363034 TI - Aging, culture and control: setting a new research agenda. AB - In the context of reviewing the current status of research on aging and control, we put forth five propositions: (1) Striving for primary control is a human universal invariant across historical time and diverse cultural settings; (2) the expression of control striving is in part shaped by culture; (3) the field needs to move away from the study of perceived control and its correlates to the study of motivational aspects of control; (4) control should be studied in a life span context and the focus should be on key transitions that redefine opportunities for control striving; and (5) inasmuch as primary control striving is such a central element of human functioning, research on its demise at the end of life should receive high priority. PMID- 10363035 TI - Perceived control as a buffer in the use of health care services. AB - Gerontologists are increasingly interested in the notion of perceived personal control because such perceptions can be threatened by age-related changes such as declining health and the loss of loved ones. Although a great deal is known about the central role of perceived control in healthy, successful aging, less is known about its potential role in specific contexts such as the use of health services. Our study examined the link between perceived control and patterns of health service use among older individuals with arthritis. We assessed perceived control during an interview, using both a domain-specific and a global measure, and considered health service use in the subsequent year. Even after statistically adjusting for age and morbidity, individuals who perceived low levels of control subsequently were found to use more health services than their high-control counterparts; they visited their physicians more often, had more laboratory tests, and stayed longer in the hospital. This was true, however, only for individuals who had also reported that their arthritis restricted the things they were able to do. Various interpretations are considered, including the possibility that patients with low perceived control are inefficient users of health services or that patients with high perceived control experience a deficiency in health care. PMID- 10363036 TI - Cognitive activity in older persons from a geographically defined population. AB - Patterns of cognitive activity, and their relation to cognitive function, were examined in a geographically defined, biracial population of persons aged 65 years and older. Persons (N = 6,162) were given cognitive performance tests and interviewed about their participation in common cognitive activities, like reading a newspaper. Overall, more frequent participation in cognitive activities was associated with younger age, more education, higher family income, female gender, and White race; participation in activities judged to be more cognitively intense was not strongly related to age, but was associated with more education, higher family income, male gender, and White race. Substantial heterogeneity in activity patterns remained after accounting for demographic factors, however. In an analysis controlling for demographic variables, level of cognitive function on performance tests was positively related to composite measures of the frequency and intensity of cognitive activity. Longitudinal studies are needed to assess the relation of cognitive activity patterns to stability and change in cognitive function in older persons. PMID- 10363037 TI - Selective interference with verbal and spatial working memory in young and older adults. AB - Young and older adults were administered digit and location memory span tasks with and without verbal and spatial secondary tasks. Age differences were greater in location span than in digit span; however, there were no age differences in either the magnitude or pattern of effects of secondary tasks. There were also no age differences in the effects of secondary tasks on a combined (digit and location) task. On the digit and location span tasks, both young and older adults showed only domain-specific interference: naming colors selectively interfered with memory for digits, leaving memory for locations unaffected; pointing to matching colors selectively interfered with memory for locations, leaving memory for digits unimpaired. The results of the present study suggest a greater age deficit in spatial working memory than in verbal working memory, but provide no evidence of an age deficit in susceptibility to interference by secondary tasks in either domain. PMID- 10363038 TI - Age differences in the strategic allocation of visual attention. AB - The allocation of visual spatial attention was investigated in two groups of adults, younger (n = 24; M = 19 yrs) and older (n = 24; M = 68 yrs). Two sequential target displays were presented on a computer screen. If a target letter appeared in Display 1, then observers were to identify a target letter in Display 2. Based on accuracy of Display 1 target detection, the older adults had a more restricted range of visual processing than the younger adults. Based on reaction times for Display 2 target identification, older adults appeared to use a spotlight (serial) scanning mechanism, whereas younger adults appeared to use an activity-distribution (parallel) mechanism. Results are consistent with age related cognitive slowing, but also suggest a difference in strategy according to the availability of visual information. PMID- 10363039 TI - Origins of individual differences in episodic memory in the oldest-old: a population-based study of identical and same-sex fraternal twins aged 80 and older. AB - The relative importance of genetic and environmental influences on episodic memory in very late life was studied using a quantitative genetic approach. Identical (n = 125) and same-sex fraternal (n = 157) twin pairs, aged 80 and older (mean age = 83.3; SD = 3.1) and without a diagnosis of dementia were tested with seven memory measures: (1-2) Digit Span Forward and Backwards, (3) Prose Recall, (4) Thurstone's picture memory test, and the Memory in Reality (MIR) test, including the subtasks of (5) free recall, (6) recognition, and (7) relocation. Heritabilities, estimated by structural equation modeling, ranged from .04 to .49. The digit span backward test showed the highest heritability (h2 = .49), while heritabilities were typically lower for the long-term memory measures. The results demonstrate genetic influences on memory in the oldest-old, but suggest that the magnitude of these effects differs across memory measures. PMID- 10363040 TI - Correlates of care quality in long-term care facilities: a multilevel analysis. AB - Hierarchical modeling was employed to explore correlates of the quality of care provided in long-term care facilities. For this purpose, a multilevel analysis offers two advantages over traditional analytical approaches. First, it accounts for the correlated nature of data recorded on multiple residents from the same facility. Second, it enables the investigators to study the influence of both resident and facility characteristics on care quality. The analysis was performed on data from 301 residents randomly sampled from 88 facilities located in the Province of Quebec, Canada. Results revealed that the presence of cognitive deficits was the strongest correlate of the quality of care provided to a resident. However, this relationship was found to vary significantly across facilities. Four facility-level variables were found to influence the relationship between cognitive functioning and care quality: the number of external collaborators the facility has, the type of training the facility manager has, the size of the facility, and the age distribution of its clientele. From these results, we suggest means to improve the quality of care provided to cognitively impaired older adults living in long-term care facilities. PMID- 10363041 TI - Sleep complaints in older women who are family caregivers. AB - Providing care to a family member with dementia has significant psychological and physical consequences. Sleep quality is likely affected by caregiving, yet this domain has received surprisingly little empirical study. In this study, sleep complaints were examined in 90 older women who were family caregivers of adults with dementia. Caregivers reported more sleep complaints than similarly aged healthy adults on all seven components of the Pittsburgh Sleep Quality Index, and a similar level of sleep complaints to those of sleep-impaired women and depressives on 6 and 4 components, respectively. Sleep medication was used by 38% of caregivers in the past month. The most common sleep complaints that occurred at least weekly were waking up in the night or early morning (84%), bathroom needs (83%), and sleep onset difficulties (41%). Sixty percent of the sample reporting nighttime care recipient disruptions stated that these disruptions occurred 3 or more times per week. Caregiver relationship and care recipient diagnosis were unrelated to sleep complaints. Lower levels of education, less internalized anger, care recipient disruptions, and psychological distress were related to poorer overall sleep quality. Sleep complaints are a common yet understudied problem in family caregivers. PMID- 10363042 TI - Effects of age and passage difficulty on listening-rate preferences for time altered speech. AB - When younger and older adults were allowed to adjust the speech rate of time compressed and time-expanded speech passages, older adults tended to select as preferred rates significantly slower speech rates than the younger adults. Both age groups, however, selected slower rates for difficult speech passages (low cloze predictability) than for easy passages (high cloze predictability). Recall performance showed effects of speech rate and passage difficulty, with participants' recall at their selected speech rates comparable to their performance at slower rates. Results suggest that older adults are as effective as the young in their ability to monitor the difficulty of a speech passage as it is being heard and to moderate their listening rate selections accordingly. PMID- 10363043 TI - Admission-related migration by older nursing home residents. AB - OBJECTIVE: To examine recent admissions of older adults to intermediate care facilities in order to identify the factors associated with whether the individual originated in another county, a non-adjacent county, and another state. METHODS: Employing a conceptual framework based upon migration theory and the long-term care decision process and a data set derived from multiple sources, logistic regression was used to examine whether characteristics of the county of residence prior to admission, the admission facility, and the individual are significant net predictors of the three types of mobility. Separate analyses were conducted for married and unmarried individuals. RESULTS: The analytical models tended to have relatively good fit but only moderate predictive accuracy. In general, persons on Medicaid payment status were more likely to move to another county and to a non-adjacent county, whereas Medicaid payment was associated with a lower likelihood of migrating to Virginia from another state. Individuals originating in counties with higher bed rates had lower rates of migration to another county and non-adjacent county, whereas those from counties with higher occupancy rates were more likely to leave their county of origin. CONCLUSION: Examination of factors associated with the distance and patterns of nursing home mobility improves our understanding of the nursing home selection process and helps to illuminate the impact of public policy, market forces, and nursing home staff on who goes where to be admitted to a nursing home. PMID- 10363044 TI - At risk on the cusp of old age: living arrangements and functional status among black, white and Hispanic adults. AB - OBJECTIVES: We examine the relationship between living arrangements and multiple measures of physical, cognitive, and emotional functioning in late midlife. METHODS: Using cross-sectional data from the Health and Retirement Study, we first assess the bivariate relationship between living arrangements and functioning; we then take into account demographic characteristics and measures of household resources and demands. RESULTS: We find evidence of differential functioning among individuals in various living arrangements. Married couples living alone or with children show the highest levels of functioning, whereas single adults living in complex households show the lowest levels. Functional deficits for those in complex households are reduced but not eliminated when we take demographic characteristics and household resources and demands into account. We find few differences by gender and race/ethnicity in the relationship between living arrangements and functioning. DISCUSSION: We show a pattern of poorer functioning among those in arguably the most demanding and least supportive household environments. This points to a vulnerable and risk-filled transition from middle to old age for these persons. Because Blacks and Hispanics show lower levels of functioning than Whites and are more likely to live in complex households, they may be particularly disadvantaged. PMID- 10363045 TI - Financial assistance from middle-aged couples to parents and children: racial ethnic differences. AB - OBJECTIVES: To examine racial-ethnic differences in the allocation of financial transfers to parents, children, and others by middle-aged couples. METHODS: Multinomial specification of alternative recipients of financial transfers, using data from the 1992 Health and Retirement Survey. RESULTS: Transfer patterns are sensitive to parental health and wealth, to children being young or in school, as well as to the donors' health and wealth. Controlling for these and other factors, including family size and structure, Blacks and Whites are the most likely, and Hispanics the least likely, to financially help their parents compared to assisting offspring. Black couples are the most likely to sacrifice their own consumption to assist parents financially. DISCUSSION: Future research on transfers should attempt to capture unmeasured noneconomic sources of variation proxied by the race-ethnicity indicator. PMID- 10363047 TI - Social networks and disability transitions across eight intervals of yearly data in the New Haven EPESE. AB - OBJECTIVES: There is considerable evidence that social networks are strongly related to survival and other health outcomes. However, findings regarding the effect of social networks on disability outcomes have been inconsistent. This study examines this relationship with respect to the risk of developing disability and recovering from disability. METHODS: Data come from a community based sample of the New Haven population aged 65 years and older, with nine annual interviews conducted between 1982 and 1991. Disability was measured by a 6 item index of activities of daily living (ADL), and a 3-item Rosow-Breslau index, with disability defined as impairment in one or more tasks on each measure. Social network variables were constructed for each of four domains of ties: children, relatives, friends, and a confidant, and a summary measure of total social networks. A Markov model was used to estimate one-year disability transitions averaged across all 8 intervals, after controlling for sociodemographic and health-related variables. RESULTS: Total social networks was associated with a significantly reduced risk of developing ADL disability (beta = -0.009, p < .01), and a significantly increased likelihood of ADL recovery (beta = 0.017, p < .01). Emotional and instrumental support did not affect the protective effect of social networks against disability, but partially accounted for their effect on enhanced recovery. Network variables related to relatives and friends were significantly associated with disability and recovery risks, but those related to children or a confidant were not. The associations with disability transitions as measured by the Rosow-Breslau index were generally smaller and nonsignificant. DISCUSSION: The findings lend further support for the role of social relationships in important health outcomes in old age. They suggest that being "embedded" in a social network of relatives and friends reduces risk for ADL disability, and enhances recovery from ADL disability. PMID- 10363046 TI - Cognitive decline and Japanese culture in a cohort of older Japanese Americans in King County, WA: the Kame Project. AB - OBJECTIVES: The prevalence of Alzheimer's disease in studies of Japanese show generally lower rates when compared with those of Caucasians. We hypothesized that among a cohort of Japanese Americans lifestyle differences would act to modify progression of the Alzheimer pathologic process over many years, resulting in a slower cognitive decline among persons whose lifestyle is more characteristically Japanese. METHODS: One thousand, eight hundred and thirty-six nondemented persons were screened with the Cognitive Abilities Screening Instrument (CASI) at baseline, and 1,604 were rescreened 2 years later. Baseline questions included migration status, exposure to Japanese culture in early life and maintenance of such culture in adulthood, and other risk factors. Cognitive decline was defined as a 2-year loss of > or = 5.15 points/100 on CASI. RESULTS: In multivariable logistic regression, variables relating to reading, writing, and speaking Japanese, being born or having lived in Japan in early life, and having friends who are only/mostly Japanese were inversely associated with cognitive decline (odds ratios ranged between 0.28 and 0.64, with p < .05). Two factors emerged in a factor analysis of these variables. The strongest explained 49% of the variance for acculturation and loaded heavily on knowledge of the Japanese language and having spent one's early years in Japan. When this factor was dichotomized into the top 20th percentile, it predicted cognitive decline with an odds ratio of 0.12 (95% CI 0.03-0.49). DISCUSSION: These results show that a Japanese lifestyle may decrease the risk of expressing cognitive decline over a 2 year follow-up period. Lower cardiovascular disease rates among Japanese may also predispose them to lower rates of cognitive decline. The greater social support characteristic of Japanese culture as well as the role that Japanese language and culture may play in neural connectivity during brain development and/or in mental stimulation in adult life may also explain our findings. PMID- 10363048 TI - Volunteering and mortality among older adults: findings from a national sample. AB - OBJECTIVES: Although a number of authors have proposed that older volunteers should benefit in terms of better health and well-being, few researchers have examined the issue empirically to see whether this is true. The purpose of this article is to build on this literature by empirically examining the association between volunteering and mortality among older adults. METHODS: Using data from a nationally representative sample, we use Cox proportional hazards regression to estimate the effects of volunteering on the rate of mortality among persons aged 65 and older. RESULTS: We find that volunteering has a protective effect on mortality among those who volunteered for one organization or for forty hours or less over the past year. We further find that the protective effects of volunteering are strongest for respondents who report low levels of informal social interaction and who do not live alone. DISCUSSION: We discuss the possibility that the curvilinear relationship we observe between volunteering and mortality is due to a combination of factors, including self-identity, role strain, and meaningfulness. Other research using more precise data is needed to determine whether these ideas are supportable. PMID- 10363049 TI - [Arterial embolization: present role of an old method of interventional radiology in the treatment of hemorrhages]. PMID- 10363050 TI - Constitutional osteochondrodysplasias identifiable at birth. A short review on the state of the art in radiodiagnosis in the late 20th century. AB - INTRODUCTION: The value of a systematic radiologic analysis in constitutional osteochondrodysplasias remains underestimated by both neonatologists and radiologists. We report the clinical experience of the Department of Neonatology and Neonatal Intensive Care Unit of St. John Hospital in Rome with constitutional osteochondrodysplasias identifiable at birth. MATERIAL AND METHODS: We reviewed 2120 cases of newborns hospitalized in our unit from January 1996 to August 1998 and here submitted to at least one direct radiograph of chest and abdomen (the so called "babygram"). All the newborns were clinically assisted by the same three neonatologists and radiologically followed by the only pediatric radiologist, the external consultant for diagnostic imaging of the Intensive Care Unit of the Neonatology Department. RESULTS, DISCUSSION AND CONCLUSIONS: We diagnosed 14 cases of constitutional osteochondrodysplasias (.66%): 4 of them (28.27%) belong to the group considered by the European Society of Pediatric Radiology (ESPR) as lethal before or immediately after birth, while the other 10 (71.43%) belong to different groups of the ESPR classification. Thus, we arbitrarily grouped them into a single pathologic condition, based on their two main features: being generally not lethal and always or very often identifiable at birth. These 10 cases were: 1 campomelic dysplasia, 3 achondroplasias, 2 asphyxiant thoracic dysplasias type Jeune, 1 cherubinic dysplasia, 2 osteogenesis imperfectae, 1 osteopetrosis. We justify the relatively high incidence of constitutional osteochondrodysplasias in our study (.66% versus an average incidence of .076% reported in the world population) on the basis of: a) an increasing number of high-risk newborns in our intensive care unit; b) an improvement in our clinical and radiologic diagnostic skills. We conclude that the state of the art of the diagnosis of constitutional osteochondrodysplasias is still based on the first plain X-ray examination performed at birth because of cardiorespiratory and/or abdominal diseases in the newborn. PMID- 10363051 TI - [Comparison of various parameters (pitch 1 and 2) in the study of the lung with spiral computerized tomography]. AB - INTRODUCTION: In Spiral CT, the pitch is the ratio of the distance the tabletop travels per 360 degrees rotation to nominal slice width, expressed in mm. Performing Spiral CT examinations with pitch 2 allows to reduce examination time, exposure and contrast dose, and X-ray tube overload. We investigated the yield of pitch 2 in lung parenchyma studies, particularly relative to diagnostic image quality. MATERIAL AND METHODS: Thirty patients were submitted to Spiral CT with pitch 1 [10 mm slice thickness, 10 mm/s table feed; 10 mm (a') and 5 mm (a") reconstruction index: protocol A] and with pitch 2 [10 mm slice thickness, 20 mm/s table feed; 10 mm (b') and 5 mm (b") reconstruction index: protocol B]. Five expert radiologists evaluated the images separately and blindly, grading noise, bronchial wall resolution and diagnostic yield on a 0-5 point scale. The results of protocol A versus protocol B images were analyzed statistically using the Mann Whitney U-test. RESULTS: The mean scores for each parameter ranged 4.13 (.70 standard deviation) for protocol B with 5 mm reconstruction index (b") to 4.81 (.44 standard deviation) for protocol A with 10 mm reconstruction index (a'). These values (max: 5) indicate very positive results on both protocol A and B images. There were no statistically significant interprotocol differences, except for bronchial wall resolution, in favor of protocol A with 5 mm reconstruction index (a") (p = .025), and for diagnostic yield, in favor of protocol A with 10 mm reconstruction index (a') (p = .018). CONCLUSIONS: Spiral CT with pitch 2 is a reliable tool for lung parenchyma studies which permits to reduce examination time and contrast dose, as well as X-ray tube overload and exposure dose. PMID- 10363052 TI - [Angiography with spiral computerized tomography in the study of aneurysms of the visceral arteries. Techniques and results]. AB - INTRODUCTION: The aneurysms of visceral vessels are characterized by few or no symptoms and the diagnosis is often occasional. We investigated the usefulness of CT angiography in the diagnosis and preoperative assessment of this condition. MATERIALS AND METHODS: From January 1993 to March 1998, twenty-five patients (aged 32-69 years) with 28 aneurysms underwent CT angiography before and after contrast agent injection to study lesion number, site, size, neck, intraluminal thrombosis and wall calcifications. Data from axial images were postprocessed on an external work-station to obtain CT angiograms. CT angiography findings were analyzed with a double blinded method by 2 radiologists comparing CT angiography with digital subtraction angiography images and evaluating the information obtained from Multiplanar (MPR), Maximum Intensity Projection (MIP) and Shaded Surface Display (SSD) reconstructions. Surgical findings were available for 10 patients. RESULTS: There was complete agreement between CT angiography and digital subtraction angiography in the identification of all lesions; the correlation rate was 94.42% for lesion location, 89.28% for lesion size, 85.71% for detection of endoluminal thrombosis and 82.14% for identification of wall calcifications. Axial and MPR images were useful in the assessment of ali parameters, while MIP images accurately demonstrated wall calcifications and the lesion relationships with adjacent structures. No additional information was obtained from SSD reconstructions. CONCLUSIONS: In our experience CT angiography can replace digital subtraction angiography in the diagnosis and preoperative work-up of visceral vessels aneurysms. CT angiography was superior to digital angiography in the evaluation of the lesion exact dimensions in cases with large thrombotic component and diffuse wall calcifications. PMID- 10363053 TI - [Quantitative assessment of portal vein flow in subjects with active chronic hepatitis: comparison of magnetic resonance angiography with bolus tracking with color Doppler ultrasonography]. AB - PURPOSE: To assess the accuracy of time-of-flight MR Angiography (MRA) with bolus tracking in evaluating mean blood velocity and flowrate in the portal vein in patients with chronic hepatitis versus healthy volunteers. MATERIAL AND METHODS: Fifteen patients with clinically-defined post-viral chronic hepatitis (viruses B and C) were examined with bolus tracking MRA and color Doppler US to evaluate portal blood flow. Both examinations were performed before and after a 1500 kcal meal. We evaluated mean blood flow velocity and flowrate in the portal vein. MRA results were compared with color Doppler findings; the results in chronic hepatitis patients were compared with those of healthy volunteers. RESULTS: The correlation between mean portal blood velocity, as measured with MRA and color Doppler US, was r = .82 before and r = .79 after the meal. There was no significant difference in mean velocity between the chronic hepatitis patients and the healthy volunteers. The correlation between portal flowrate, as measured with MRA and color Doppler US, was r = .87 before and r = .91 after the meal. There was no significant difference in mean flowrate between the chronic hepatitis patients and the healthy volunteers. In contrast, there were significant differences in mean velocity and portal flowrate, as measured with MRA before the meal, between the chronic hepatitis patients and the healthy volunteers. DISCUSSION AND CONCLUSIONS: Bolus tracking MRA is superior to color Doppler US in quantitating blood flow in the portal vein and evaluating changes after a meal. Decreased mean velocity and flowrate may indicate impaired function, as it happens in early chronic hepatitis. PMID- 10363054 TI - [Contrast media extravasation in upper abdominal injuries: detection with spiral computerized tomography]. AB - PURPOSE: The possibility of detecting contrast agent extravasation (i.e., active hemorrhage) with dynamic conventional Computed Tomography (CT) in patients with abdominal trauma has already been reported in small series. We report our experience in the demonstration of contrast material extravasation using helical CT; we also investigate the diagnostic and clinical value of this finding. MATERIAL AND METHODS: January 1997 to July 1998, we examined 41 consecutive patients with upper abdominal trauma. Twelve patients (29%) had contrast material extravasation. The examinations were performed with a helical unit and volumetric acquisitions (thickness 8-10 mm, pitch 1, reconstruction interval 5-8 mm). The intravenous contrast medium (350 mgI/mL, 130-140 mL) was administered with rapid infusion (2-2.5 mL/s, 40-50 s acquisition delay from bolus starting) and using a power injector. We reviewed the CT studies and clinical records of these 12 patients. Contrast agent extravasation was considered present when this finding, not recognizable on plain scans, showed equal attenuation to or higher attenuation than the vessels within the same level. Moreover we assessed leak site, CT appearance, the direct visualization of the involved vessel, the evidence of other abdominal or extra-abdominal injuries, the CT signs of hypovolemic shock, clinical and surgical data. For comparison, we finally evaluated 50 examinations performed with a conventional CT scanner in subjects with abdominal trauma. RESULTS: Active hemorrhage involved the abdominal wall in 1 case (intercostal artery), the solid organs in 4 (splenic in 2, hepatic in 1, of the middle hepatic vein in 1), the peritoneal cavity in 3 (splenic, midcolic, and gastroduodenal artery in 1 each), the retroperitoneum in 4 (renal pedicle in 2, renal parenchyma in 1, lumbar artery in 1). In all cases the site of contrast extravasation corresponded at surgery to the site of active bleeding. The pattern was localized in 10 cases and diffuse in 2. The involved vessel could be identified in 5 cases while in the other ones the origin could be inferred from the leakage site. Associated injuries of upper abdominal organs were seen in 11 of 12 patients and extra-abdominal trauma in 6. In 4 cases there were CT features of hypovolemia. One patient died during transport to the operating room and another after surgery, while all the others survived. Contrast extravasation was identified in 9 (18%) of the patients examined with a conventional CT unit. CONCLUSIONS: Active contrast material extravasation can be recognized with conventional CT scanners, though it has been considered a rare finding. Helical CT seems to increase the detection rate and especially to boost the radiologist's confidence in this diagnosis. Though active bleeding is identified in severely injured subjects requiring urgent intervention and may be associated with findings of hypovolemic shock, it should not be considered itself as a negative prognostic factor. Contrast extravasation is due to ongoing hemorrhage and its detection is critical for urgent treatment. Accurate anatomical location permits to choose surgical management or transcatheter embolization and thus decreases time consumption for precise bleeding site identification. PMID- 10363056 TI - [Doppler color in superficial adenopathies]. AB - INTRODUCTION: Superficial lymph nodes are frequently involved in different diseases. Their location makes them suitable for effective assessment with high resolution US and color Doppler has been recently suggested as a tool for increasing sensitivity in lymph node studies. Thus, we investigated the main vascular patterns detectable in abnormal superficial lymph nodes. PATIENTS AND METHODS: We evaluated 260 nodes in 180 adult patients; the nodes were located in the cervicofacial ring (78, 30%), internal jugular stations (104, 40%), and supraclavicular (44, 17%), axillary (21, 8%), and inguinal (13, 5%) stations. Color Doppler was performed with 7.5-13 MHz linear transducers, with parameters adjusted for slow-flow detection (5-6 MHz frequency, 700-900 Hz PRF, 50 Hz band filter, high color persistence). Disease assessment required fine-needle biopsy (95 nodes in 95 patients) and clinical follow-up (165 nodes in 85 patients). RESULTS: Fifty-five nodes (21%) presented acute and 130 (50%) chronic inflammation: 75 nodes (29%) were metastatic. The following vascular patterns were detected: a single vascular pole (type I) was seen in chronic inflammation (72% sensitivity, 86% specificity, 57% positive and 92% negative predictive value); an enlarged single vascular pole, with 2-3 enlarged branches (type II) in acute adenitis (80% sensitivity, 81% specificity, 78% positive and 83% negative predictive value); multiple vascular poles with many deformed and displaced branches converging centrally (type III) in metastases (76% sensitivity, 100% specificity, 100% positive and 91% negative predictive value). CONCLUSIONS: We conclude that color and power Doppler are useful integrations to B-mode US because they can detect specific signs of malignancy such as peripheral vascular poles and intranodal displacement of vessels. PMID- 10363055 TI - [Role of color Doppler ultrasonography with contrast media in the monitoring of hepatocarcinoma after intralesional treatment]. AB - INTRODUCTION: We investigated the accuracy of contrast-enhanced color Doppler US in the assessment of the effectiveness of intralesional treatment of hepatocarcinomas. MATERIAL AND METHODS: Eight cirrhotic patients (HCV+), Child Pugh class B, with a single hepatocarcinoma (< 4 cm O) and ineligible for surgical resection for various reasons (age > 70 years, reduced partial hepatic reserve, esophageal varices at risk, postoperative recurrence, no consent to the operation) were submitted to radiohyperthermia (6 patients) and percutaneous alcoholization (2 patients). The diagnosis was made with alpha-fetoprotein titration. CT, B-mode and color Doppler US with the administration of Levovist (Schering AG, Berlin, Germany). Thirty and 60 days after the treatment, both the alpha-fetoprotein titration and contrast-enhanced color Doppler US were repeated. RESULTS: Baseline color Doppler was carried out before intralesional treatment in the 8 patients and was followed by Levovist color Doppler which showed some intralesional signals, afferent vessels and rich vascularization in all the lesions. At the first follow-up (30 days), no intralesional vascular signals or afferent vessels were detected in any patient, while rich peripheral vascularization persisted in all cases, even after radiofrequency and alcoholization treatments. At 60 days' follow-up, the color Doppler pattern of all cases was the same as at 30 days. CONCLUSIONS: The absence of any intralesional vascular signals in all the treated patients and the possible demonstration of complete tumor necrosis seem to confirm the important role of contrast-enhanced color Doppler US in monitoring focal hepatic lesions after intralesional treatment. PMID- 10363057 TI - [Transluminal treatment in aneurysm of the abdominal aorta. Role of diagnostic imaging]. AB - PURPOSE: To emphasize the importance of diagnostic imaging: a) in the selection of patients to be treated with the transluminal approach b) during stent-graft positioning c) in the follow-up of treated patients. MATERIAL AND METHODS: From January 1997 to May 1998, twenty-five patients with abdominal aortic aneurysms (AAA) were treated with transfemoral stent-grafts (AneuRx Medtronic). All patients were submitted to a preoperative study including digital angiography (DSA) and Spiral CT. Intraoperatively they underwent DSA and intravascular ultrasound (IVUS). Follow-up was performed with Spiral CT. RESULTS: Twenty-four bifurcated and one straight device were inserted in twenty-five patients with AAA. In two cases it was impossible to position the endoprosthesis due to narrow or tortuous artery access. Following the intention to treat criteria, the technical success rate was 92.5%. All the stents were patent and no dislodgement was observed at follow-up. Partial thrombosis of the stent was observed in three patients. Owing to incomplete distal covering, early endoleak occurred in one patient with an aortoiliac aneurysm; the positioning of two cuffs allowed a successful outcome. CT examination performed 6 months after positioning revealed the presence of endoleaks in three patients, due to persistence of lumbar and inferior mesenteric artery patency. The AAA was no more appreciable in five of the ten patients submitted to CT follow-up one year after the procedure. In three of ten cases it was reduced in size and in two patients there was no change. DISCUSSION: Spiral CT plays a basic role in the selection of patients because it helps assess the length and diameter of the proximal neck, thus permitting to choose the device to be inserted. Preoperative DSA is mandatory in the evaluation of size and tortuosity of the iliac arteries. IVUS allows to monitor the site of delivery during the maneuver and to make the final measurements while DSA plays a role in checking the correct positioning of the device and excluding the presence of endoleaks at the end of the procedure. Late follow-up with Spiral CT aims at demonstrating possible malfunctioning of the endoprosthesis and confirms the definitive exclusion of AAA. CONCLUSIONS: Diagnostic imaging plays a basic role in the endovascular treatment of AAA, much more than that required for traditional surgical treatment. In particular, pretreatment planning is critical and requires sophisticated imaging including Spiral CT with 3D reconstruction and angiographic evaluation using catheters with calibrated markers. PMID- 10363058 TI - [Computerized tomography-guided drainage of postoperative abdominal fluid collections]. AB - INTRODUCTION: We report our personal technique and the results of CT-guided percutaneous drainage of postoperative abdominal fluid collections. MATERIAL AND METHODS: January 1990 to March 1998, eighty-three patients were treated for postoperative abdominal fluid collections. Forty-eight patients had undergone bowel resection, 11 laparoscopic cholecystectomy, 3 cholecystectomy, 5 splenectomy, 3 cephalopancreasectomy, 6 hepaticojejunal anastomosis, 4 hepatic resection, 2 laparocele, 1 hysterectomy. The complications had developed few days to about one year postoperatively. The suspicion of abdominal fluid collection was supported by clinical and laboratory findings. All patients were submitted to a preliminary CT scan to locate the fluid collection, assess its morphology and relationships with surrounding structures, and plan the safest access route. After local anesthesia, a trial fine needle (Chiba 20-22 G) aspiration was performed and then the draining tube was inserted into the collection under CT guidance; the tube caliber depended on the fluid amount and viscosity. After drainage, the tube was removed if CT showed complete resolution of the fluid collection; otherwise it was left in place for subsequent washing of the cavity. Based on clinical, laboratory and CT findings, another CT-guided percutaneous drainage was judged necessary in 30 patients, 2-9 days after the first one. Drainage was considered successful when sepsis resolved and no further percutaneous/surgical drainages were needed. RESULTS: CT-guided percutaneous drainage was successful in 61 of 83 patients (73.5%); the fluid collection resolved after one drainage in 26/61 patients, in 2-9 days in 18/61, and after a second CT-guided drainage in 17/61. Drainage was not resolutive in 22 of 83 patients, because major postoperative complications required laparotomic surgery; these complications were fistulas (anastomotic in 12 cases; pancreatic in 5 and biliary in 3) and segmentary bowel necrosis in 2 cases. Intracavitary bleeding as a catheter-related complication occurred only in one patient with an anterior abdominal wall abscess. CONCLUSIONS: CT-guided percutaneous drainage offers many advantages over surgery: it is less invasive, can be repeated and requires no anesthesia; there are no surgery-related risks and lower morbidity and mortality rates. Moreover, subsequent hospitalization is shorter and costs are consequently reduced. We conclude that CT-guided percutaneous drainage is the method of choice in the treatment of postoperative abdominal fluid collections. PMID- 10363059 TI - [Cost analysis in diagnostic senology]. PMID- 10363060 TI - [Unusual application of computerized tomography: the study of musical instruments]. AB - INTRODUCTION: We report on the use of CT in the study of bowed stringed instruments to assess structural defects and/or damage before proceeding to any repair. MATERIAL AND METHODS: Two antique masterpieces from the Castello Sforzesco Museum of Antique Musical Instruments were analyzed with CT. They were an exquisite wood and ivory guitar from Naples (Italy) and a very rare Giuseppe Guarneri "del Gesu" violin from Cremona (Italy), both crafted in the early years of the 18th century. We evaluated the wood thickness, the neck and its heel. In the wood structure we studied the course and thickness of hypo- and hyperdense lines. RESULTS: The examination showed three types of signs: normal wood structure: hypodense, thin, parallel lines; wormholes: hypodense lines with irregular course and variable thickness; previous repair signs: thin or thick more or less parallel hyperdense lines. CONCLUSIONS: The study confirmed that CT is a valuable tool to investigate normal structure, defects and damage, providing accurate information for the evaluation and repair of antique stringed instruments. PMID- 10363061 TI - [Biologic effects of the static magnetic field generated by a 0.5 T magnetic resonance tomograph on the enzyme activity of catalase and creatine kinase in the rat]. AB - PURPOSE: We investigated possible alterations in the enzyme activity of catalase and isozyme MB-creatine kinase induced by prolonged exposure of laboratory rodents to a static magnetic field generated by a .5 T Magnetic Resonance unit. MATERIAL AND METHODS: Thirty Wistar albino mice were divided into two groups of 15 mice, one to be exposed to the static magnetic field for 12 hours and the other to be kept in the same environmental conditions as a control group. Immediately after the exposure a peripheral venous blood sample was collected, the cardiac muscle was removed from the mice and the enzyme activity of catalase and MB-creatine kinase were assayed using the spectrophotometric analysis. RESULTS: No statistically significant variation was detected between the enzyme activity of catalase and MB-creatine kinase in the serum and cardiac muscle of the exposed versus the control mice. In the mice exposed to the static magnetic field the enzyme activity of serum and cardiac muscle catalase were respectively .2154 U/L and .0707 U/L after 10 minutes; they were; .2699 U/L and .0946 U/L after 160 minutes. In the control mice the enzyme activity of serum and cardiac muscle catalase were respectively .1941 U/L and .0707 U/L after 10 minutes; they were .2061 U/L and .1068 U/L after 160 minutes. The enzyme activity of MB creatine kinase in mice was measured in the exposed (80.8 U/L) versus the control (79.6 U/L) group: the difference does not exceed standard deviation. DISCUSSION AND CONCLUSION: Our results seem to exclude any alteration in the activity of catalase and MB-CK after 12 hours' exposure to the static magnetic field. However some homeostatic mechanisms peculiar to pluricellular organisms might act in vivo to adapt to the effects of the static magnetic field during exposure. PMID- 10363062 TI - [Performance assessment of mammographic diagnostic systems: evolution of methods and their application to a digital image study]. AB - INTRODUCTION: "Receiver Operating Characteristic" (ROC) curves are one of the most efficient analysis tools for the complete evaluation of a diagnostic system performance. However this method is limited in visualizing and locating abnormal structures, such as clusters of microcalcifications on mammographic images. Other more refined and complex techniques have also been suggested, where particular statistical hypotheses are assumed, namely the "Free-response ROC" (FROC), the "Alternative FROC" (AFROC) and the "Free-response Forced Error" (FFE) analyses. We studied the theoretical bases of these different methods and their experimental applications to assess the correctness of the hypothetical statistical distributions. MATERIAL AND METHODS: We considered two statistical hypotheses: first, that the false-positive response distribution follows the Poissonian statistics; second, that "signal" and "noise" distributions have a Gaussian trend with different means and variances. Thus, we applied the different methods to the responses given by 8 observers (5 radiologists and 3 medical physicists) who independently evaluated 3 digital mammographic samples. Every sample consisted of 39 images, with 1-15 clusters each (total: 100 clusters). The samples were obtained from 39 images available in an Internet database (sample 1); 2 different digital filters were applied to each image (samples 2 and 3). To collects responses, we provided for two phases: first, every observer visualized and located the clusters at a given confidence level; second, when a false positive response was given, spontaneously or after forcing, the responses were ordered by decreasing conspicuity. Finally, data were analyzed with a "home-made" software by applying the FROC and AFROC analyses to the data collected in phase 1 and the FFE analysis to those collected in phase 2. RESULTS: We considered the area under the AFROC curve as the most important parameter: the values obtained with the 3 types of analysis are well in agreement within their uncertainties. In particular, the FROC-AFROC agreement did not exceed 5.9% (10 of 14 cases within 2.5%), while the FFE analysis had higher standard deviations associated with the area value (about 10%). The interpolated curves from both FROC and AFROC data were very similar. The three methods had various advantages: the FFE is very simple to calculate and makes the most of the information given by the observer; FROC and AFROC can provide true-positive and false-positive responses on the same image, which permits to optimize the evaluation of a diagnostic system performance. The statistical tools used in the simplest methods are usually integrated with the completeness characteristics of the location of multiple signals on mammograms. CONCLUSIONS: In theory, every method is necessary because it provides additional information to validate the statistical hypotheses under investigation. In fact, when the methods are used to evaluate and compare several diagnostic systems, the results of the three techniques are equivalent. Therefore, choosing a specific technique depends on both available resources and response type all the hypothetical statistical distributions in our study proved correct. PMID- 10363063 TI - [A case of atypical stress fracture]. PMID- 10363064 TI - [Villonodular synovitis of the hip. Combined diagnostic imaging of a case]. PMID- 10363065 TI - [Chlamydia pneumoniae pneumonia. Radiographic features in a patient with lung transplantation]. PMID- 10363066 TI - [Giant Spigelian hernia: assessment with computerized tomography and surgical correlations. Report of a case]. PMID- 10363067 TI - [Cystic duplication of the stomach. Ultrasonography compared with CT and MRI in a case]. PMID- 10363068 TI - [Magnetic resonance diagnosis of large cystic lymphangioma of the spleen]. PMID- 10363069 TI - [Cholangiography with ultrasonography and contrast media in the diagnosis of obstructive jaundice caused by ++Klatskin tumor. Report of a case]. PMID- 10363070 TI - [Monitoring of ureteral stent color Doppler ultrasonography. Report of a case]. PMID- 10363071 TI - [The lady of Via della Signora]. PMID- 10363073 TI - Patient compliance with quality of life questionnaires. Italian Group for Evaluation of Outcomes in Oncology (I.G.E.O.). AB - AIMS AND BACKGROUND: To evaluate the rate of cancer patients who do not fill out a quality of life (QL) questionnaire, their characteristics and the reasons for not filling out the QL questionnaire. METHODS: Consecutive cancer patients who were seen in 79 Italian medical oncology and radiotherapy centers over a period of one week were asked to fill out a questionnaire concerning the importance of 46 domains of quality of life, each one scored on 4 levels (not at all, a little, much, and very much). RESULTS: Of 6,918 cancer patients, 820 (11.9%) did not fill out the questionnaire. The most important reasons for not complying were: illiteracy (17.9%), lack of glasses or poor eye-sight (17.4%), poor physical condition (11.9%), poor psychological condition (5.9%), refusal (28.7%). The questionnaires significantly less filled out were those of older patients with low performance status and educational level or with locally advanced or disseminated disease and inpatients. CONCLUSIONS: The results of the study reveal the risk of selection bias in QL assessment in randomized controlled trials and suggest the need for more complete information regarding the aim of QL evaluation and the necessity of a proxy's help to overcome the problem, with the awareness that the proxy's influence could modify the response. The impact of the lack of patient compliance on the QL results still remains to be evaluated. PMID- 10363072 TI - Protection of normal tissues from radiation and cytotoxic therapy: the development of amifostine. AB - Much effort is being made to reduce the iatrogenic toxicity of antineoplastic treatments in order to improve the quality of life of cancer patients. Cytoprotection of healthy tissue by thiol group donors is one of the most promising lines of research. Amifostine is the most extensively studied drug in the category. We reviewed the extensive medical literature on amifostine. The protective effect of amifostine has been demonstrated for cisplatin-induced toxicity in lung and ovarian cancer, with particular regard to nephrotoxicity, neurotoxicity and neutropenia. No protective effect has been seen for tumor cells owing to a selective action of amifostine on healthy tissues. A frequent side effect of amifostine is a transient decreases in blood pressure; it is usually asymptomatic if an easily handled premedication is given. Cytoprotection by amifostine is also well known for alkylating drugs and radiation therapy, whereas it is still the object of study for new drugs, especially taxanes. The present work also includes a cost-benefit analysis and a prospective view on the most promising research lines. PMID- 10363074 TI - A double-blind evaluation of the analgesic efficacy and toxicity of oral ketorolac and diclofenac in cancer pain. The TD/10 recordati Protocol Study Group. AB - AIM: To compare the analgesic efficacy and toxicity of the nonsteroidal anti inflammatory analgesic drug, ketorolac (Toradol, Recordati spa, Milan) 10 mg p.o. (t.i.d.) with diclofenac (Voltaren, Novartis Farma, Origglo, VA) 50 mg p.o. (t.i.d.) in cancer patients with moderate to severe chronic pain. METHODS AND STUDY DESIGN: The study was a multicenter randomized double-blind cross-over trial. Each treatment lasted 7 days, after which the patients crossed over to the other drug. Pain intensity was evaluated by the visual analogue scale (VAS) after the first dose and by the 5-point verbal rating scale (VRS) by the patient and by the physician following the 7-day treatment. RESULTS AND CONCLUSIONS: A total of 138 advanced cancer patients were enrolled in the study. Overall 251 single-dose administrations (117 cross-over observations) and 257 multiple treatments (127 cross-over experiments) were assessable. After a single administration of ketorolac and diclofenac, no significant difference could be observed in analgesic activity, as indicated by the area under the pain-intensity time curve (AUC0-8), in the maximum efficacy, or the duration of efficacy of the two drugs. The Westlake confidence intervals of the AUC0-8 ratio (ketorolac: diclofenac) (1.07; 90% CI, 0.94-1.19), of the maximum efficacy ratio (1.03; 90% CI, 0.92 1.14), and the duration of efficacy ratio (1.05; 90% CI, 0.97-1.11) showed the bioequivalence of the two drugs. Satisfactory pain relief was reported for multiple 7-day treatments, with no significant differences between the two therapies: according to the physician's evaluation, in 93/128 (73%; 95% CI, 65 80%) ketorolac treatments and 91/129 (71%; 95% CI, 63-78%) diclofenac treatments; according to the patient's evaluation, in 83/128 cases (65%; 95% CI, 57-73%) after ketorolac and in 74/129 cases (57%; 95% CI, 49-66%) after diclofenac. Adverse symptoms were acceptable with both drugs. Interestingly, a pronounced sequence effect was found: gastric disturbances after ketorolac were observed mainly (10 out of 15 observed events) when the drug was given to patients pretreated with diclofenac. PMID- 10363075 TI - Prognostic value of clinicopathologic characteristics including neuroectodermal differentiation in osseous Ewing's sarcoma family of tumors in children. AB - AIMS AND BACKGROUND: The aim of the present study was to determine the relationship between clinico-pathologic parameters, including neuroectodermal differentiation, and their impact on survival in a series of pediatric patients with osseous tumors of the Ewing's sarcoma family admitted to the Pediatric Department of the Istituto Nazionale Tumori of Milan. METHODS: Seventy-three patients were enrolled. The variables analyzed were sex, age, site of primary tumor, serum lactate dehydrogenase (LDH) level at diagnosis, involvement of periosseous soft tissues by primary tumor, presence of metastatic disease, status of disease after the treatment plan, as well as the presence of mitoses, morphologic and immunocytochemical neural markers, and neuroendocrine markers in the primary tumor. RESULTS: Neural and neuroendocrine markers were not significantly associated with any of the other parameters. In the univariate analysis, significant risk factors related to unfavorable outcome were elevated LDH, metastatic disease, lack of complete remission after treatment, presence of mitoses and of morphological neural markers; immunocytochemical neural and neuroendocrine markers lacked prognostic value. In the multivariate analysis, only LDH levels and the status of disease following the treatment were retained. CONCLUSIONS: LDH level at diagnosis might be a useful marker to identify different risk levels; neuroectodermal differentiation might have no clear-cut impact on the clinical management of osseous Ewing's sarcoma family of tumors. PMID- 10363076 TI - N-nitroso compounds and Helicobacter pylori in the gastric remnant. AB - AIMS AND BACKGROUND: The aims of this study were to investigate the role of N nitroso compounds (NOC) and Helicobacter pylori (H. pylori) in gastric stump carcinogenesis. METHODS AND STUDY DESIGN: Analyses of biochemical parameters such as pH and NOC concentration were carried out on 65 fasting gastric juice samples obtained at endoscopy from 45 patients previously submitted to partial gastrectomy for benign peptic ulcer disease (23 Billroth I, 22 Billroth II/Reichel-Polya) and 20 normal controls. Biopsy specimens were taken to determine histology and H. pylori status. RESULTS: Significantly higher mean pH values and NOC concentrations were found in partial gastrectomies compared to normal controls. In relation to surgical methods, higher mean pH values and NOC concentrations were observed in the gastric juice of patients with Billroth II compared to Billroth I gastrectomies. Independently of the type of surgical reconstruction, higher mean NOC levels were recorded in patients with more severe histological changes and H. pylori infection. CONCLUSIONS: All these data suggest that high levels of NOC in gastric juice and H. pylori infection could be cofactors in gastric stump carcinogenesis. PMID- 10363077 TI - Gastric non-Hodgkin's lymphoma: analysis of 252 patients from a multicenter study. AB - AIMS AND BACKGROUND: The stomach is the most common site of primary extranodal non-Hodgkin's lymphoma (NHL) and no agreement has been reached so far on the best therapeutic approach. The main objects of this study were to report the long-term results and to evaluate the importance of some possible prognostic factors in a large series of patients. NHL was considered primary gastric if the main symptoms at presentation were those of gastric disease. METHODS AND STUDY DESIGN: We analyzed 252 consecutive patients treated between 1980 and 1993 in five hospitals in north-east Italy. According to the Working Formulation, 98 patients had low grade lymphoma, 59 intermediate grade (D to F), 81 G or high grade and 14 were not classified. The patients were divided into two groups: one including patients with limited disease (localized to the stomach or perigastric lymph nodes: 165 patients) and one including those with advanced disease (87 patients). The treatment consisted of surgery, chemotherapy, radiotherapy or combinations of these. Sixteen patients received only supportive therapy. RESULTS: The five-year overall survival was 65.4%: 80.3% for patients with limited disease and 36.7% for those with advanced disease (P < 0.0001). Among the limited disease patients the five-year survival was 84.4% for those treated with gastrectomy alone and 88.7% for those who received also adjuvant chemotherapy (P = 0.11). However, while chemotherapy did not improve survival in low grade NHL, it seemed to produce a better survival in the intermediate and high grade groups (P = 0.06). Twelve patients were treated with primary chemotherapy and the five-year survival was 71.2%. In multivariate regression analysis the most important variable for overall survival was surgery for the whole group of 252 patients (P < 0.0001), while it was age for the group with limited disease (P = 0.0008). CONCLUSIONS: Surgery alone can be curative for most patients with gastric lymphoma limited to the stomach or to the perigastric lymph nodes; surgery followed by chemotherapy seems to produce better results than surgery alone in intermediate and high grade lymphomas. Also a non-surgical approach with first-line chemotherapy is associated with a high rate of complete remissions and five-year survival. In advanced disease the five-year survival is similar to that of nodal NHL. PMID- 10363078 TI - Clinical approach in patients with metastatic differentiated thyroid carcinoma and negative 131I whole body scintigraphy: importance of 99mTc MIBI scan combined with high resolution neck ultrasonography. AB - AIMS AND BACKGROUND: The aim of this study was to define the clinical impact of MIBI scan combined with neck ultrasonography on the detection of metastates in differentiated thyroid carcinoma (DTC) patients with elevated serum Tg levels but negative 131I scan (non-functioning DTC). METHODS AND STUDY DESIGN: Eighty-two patients with non-functioning DTC, 19 patients with 131I-positive metastases (functioning DTC), and 24 DTC patients who were disease free after therapy (no cancer patients) were enrolled. 131I scan was performed after administration of low diagnostic and high therapeutic tracer doses. Early and delayed images were obtained after MIBI injection. Neck-chest CT scan and/or MRI were also performed in patients with non-functioning DTC. RESULTS: In the group of non-functioning DTC patients, metastatic foci were detected in 71/82 cases: in the cervical lymph nodes in 51 cases (sensitivity 94.1% with MIBI, 90.2% with US, 35.3% with CT/MRI), mediastinal lymph nodes in 31 cases (sensitivity 100% with MIBI, 58% with CT/MRI), lungs in 8 cases (sensitivity 100% with both MIBI and CT/MRI), and bone in 2 cases (sensitivity 50% with MIBI, 100% with MDP bone scan). Among the 19 patients with functioning DTC a close relationship between MIBI and 131I findings was observed. As regards the 24 tumor-free patients, MIBI was correctly negative in all cases, while US visualized enlarged cervical lymph nodes that were suspected to be neoplastic but proved to be inflammatory lesions at cytology in three patients. CONCLUSIONS: On the basis of these data, MIBI scan combined with neck US could be proposed as a first-line diagnostic imaging modality in the follow-up of DTC patients with elevated serum Tg levels and negative 131I scan. PMID- 10363079 TI - Relationship between the shape and size of radiofrequency induced thermal lesions and hepatic vascularization. AB - AIMS AND BACKGROUND: The aim of this study was to evaluate the relationship between hepatic vascularisation and the final size and shape of radiofrequency (RF) induced thermal lesions. METHODS: Series of four RF thermal lesions were created in explanted calf livers and in pig livers maintaining the following experimental conditions throughout the procedure: normal hepatic perfusion, occlusion of the hepatic artery, occlusion of the portal vein, occlusion of both hepatic artery and portal vein (Pringle maneuver) and subtotal occlusion of the hepatic veins. A 14G expandable needle electrode was used to make the thermal lesions. Each lesion was created applying predetermined temperatures ranging between 95 and 115 degrees C and an exposure time of 20 minutes. RESULTS: Occlusion of the hepatic artery during the RF procedure resulted in moderate and not significant increases in thermal lesion diameter compared with those obtained in normally perfused liver (3.0 +/- 0.4 cm vs 3.0 +/- 0.2 cm), while occlusion of the portal vein resulted in larger lesion diameters (3.5 +/- 0.3 cm). In both these cases the diameters of the thermal lesions were smaller than those obtained in explanted calf liver (4.0 +/- 0.3 cm) and their shape showed peripheral irregularities. Thermal lesions larger than those seen in normally perfused liver and equaling those observed in explanted calf liver were created both during the Pringle maneuver (4.0 +/- 0.2 cm) and after subtotal occlusion of the hepatic veins (4.0 +/- 0.3 cm). In both these cases the thermal lesions were regular in shape. CONCLUSIONS: Occlusion of the blood flow during the RF procedure avoids heat loss by convection, resulting in the creation of larger thermal lesions than those obtained in normally vascularized liver using the same electrode, temperatures and exposure time. This technique could therefore be employed in humans to destroy large hepatic tumor nodules. PMID- 10363080 TI - Stromal osseous metaplasia in metastatic adenocarcinoma of the gallbladder. AB - A case of stromal osseous metaplasia in the abdominal scar metastasis of a gallbladder adenocarcinoma is described. The occurrence of stromal osseous metaplasia in carcinomas probably does not affect prognosis; however, it must be recognized to avoid a misdiagnosis of carcinosarcoma. PMID- 10363081 TI - Recurrent primary well-differentiated intrascrotal liposarcoma: case report and review of the literature. AB - Lipoma-like liposarcomas of the scrotal wall are very rarely reported neoplasms in the surgical and histopathological literature. We treated a well differentiated liposarcoma of the inside wall of the scrotum in a 62-year-old man. Following local excision, the tumor recurred after three months, and a funiculoorchidectomy was performed. Today, 24 months following secondary surgery, the patient is completely asymptomatic and there is no evidence of tumor recurrence either on physical examination, ultrasonography or abdominal and pelvic computed tomography. In this paper we present the case and a review of the relevant literature. PMID- 10363082 TI - Long-term survival of a patient with a giant-cell tumor of the sphenoid body: a case report. AB - Giant-cell tumors (osteoclastomas) of the sphenoid body are rare, histologically benign lesions that can grow in different directions within the bony structures of the skull base. To date, the precise role of the different surgical strategies (radical versus partial resection; biopsy) and of supplementary postoperative radiotherapy remains undefined due to the short follow-up of the majority of the reported cases. We present a patient who is alive without symptoms or signs of recurrence 44 years after a macroscopically radical excision of the neoplasm followed by radiotherapy. PMID- 10363083 TI - Lung and aero-digestive cancers in young marijuana smokers. AB - Marijuana has been shown to be one of the commonly abused substances in the world, especially among teenagers and young adults. Although its addictive potential and psychomotor side-effects have been widely publicized, the issue of possible carcinogenicity is not as well perceived. Marijuana smoke contains many of the same organic and inorganic compounds that are carcinogens, co-carcinogens, or tumor promoters found in tobacco smoke. We have encountered several young marijuana users with no history of tobacco smoking or other significant risk factors who were diagnosed to have lung or other aero-digestive cancers in our practice. Although there are several experimental and epidemiological studies suggesting an association of marijuana use as a possible cause of cancers, this issue remains controversial. It is hoped that our case presentation can help to stimulate further awareness and research into this issue. PMID- 10363084 TI - Minimal requirements in prostate cancer irradiation: a consensus document by the AIRO Lombardia Cooperative Group. AB - PURPOSE: With the aim of establishing clinical and technical criteria to homogenize radiotherapy practice, a working group of AIRO-Lombardia (Associazione Italiana di Radioterapia Oncologica--Gruppo regionale della Lombardia) has tried to define minimal requirements for radical and postoperative irradiation in prostate cancer. The document has been structured in such a way as to be also of interest to the urological and medical oncology communities. METHODS: The working group, composed of representatives of most of the regional radiotherapy departments in the Lombardy region, had monthly meetings during 1996 and 1997. The document on minimal requirements has been derived from the participants' combined experience and knowledge, from review of the literature, and from a 1995 regional survey on current practice of prostate irradiation. RESULTS: Minimal requirements for radical and postoperative irradiation of prostate cancer have been defined with respect to treatment strategies, pre-treatment diagnostic evaluation and staging, treatment prescription, preparation and execution, and quality assurance procedures. CONCLUSION: Standards of reference for minimal requirements in prostate cancer irradiation adapted to the regional structures and resources have been defined. PMID- 10363085 TI - Tumori. Chronotherapy Study Group of the European Organization for Research and Treatment of Cancer. PMID- 10363086 TI - HSUS sues FDA to end preventive extralabel use. PMID- 10363087 TI - Applauds increased regulation of antimicrobials. PMID- 10363088 TI - Controversy over pet insurance. PMID- 10363089 TI - Controversy over pet insurance. PMID- 10363090 TI - What is your diagnosis? Bilateral increased uptake of technetium Tc 99m medronate in the region of the tarsal joints. PMID- 10363091 TI - Diagnosis and treatment of copper toxicosis in ruminants. PMID- 10363092 TI - The duty of a veterinary medical association to discharge members. PMID- 10363093 TI - Proposals for increasing the availability of animal drugs for minor species and minor uses. PMID- 10363094 TI - Minor-use animal drug program--a national agricultural program to approve animal drugs for minor species and uses. PMID- 10363095 TI - Wound contamination and antimicrobial susceptibility of bacteria cultured during total ear canal ablation and lateral bulla osteotomy in dogs. AB - OBJECTIVE: To detect contamination of wound sites from surgical handling of excised tissues during total ear canal ablation and lateral bulla osteotomy in dogs, and to compare susceptibility of bacterial isolates to cefazolin with susceptibility to other antimicrobial agents. DESIGN: Prospective clinical study. ANIMALS: 13 dogs. PROCEDURE: Dogs were treated surgically for otitis externa and media via total ear canal ablation and lateral bulla osteotomy. Specimens for aerobic bacterial culture were obtained from SC tissue immediately following skin incision, tissues excised from the osseous bulla (after transection of the horizontal ear canal and lateral bulla osteotomy), and from SC tissue prior to skin closure. Antimicrobial susceptibility of bacterial isolates to various antibiotics was determined by use of a broth dilution assay. RESULTS: There was a significant association between isolation of Streptococcus canis and Escherichia coli from specimens from the osseous bulla and specimens from the SC tissues prior to skin closure, indicating contamination of the SC tissues during surgery. Seventy percent of bacterial isolates were susceptible to cefazolin. CLINICAL IMPLICATIONS: Measures to limit bacterial contamination resulting from tissue handling during total ear canal ablation and lateral bulla osteotomy are necessary. Bacteriologic culture of tissue of the osseous bulla and determination of antimicrobial susceptibility are recommended. Administration of cefazolin alone may not be efficacious for antimicrobial prophylaxis. PMID- 10363096 TI - Sino-orbital aspergillosis in a dog. AB - Fungal rhinitis-sinusitis with orbital involvement was diagnosed in a dog with profuse unilateral ocular and bilateral nasal discharge, enophthalmos, and a corneal descemetocele. The descemetocele was treated with a conjunctival pedicle graft. Frontal sinusotomy was performed, and clotrimazole was infused through catheters placed in the frontal sinuses and nasal cavities to saturate the tissues for 1 hour. Successful resolution of orbital infection may have been aided by a fistula between the frontal sinus and orbit that allowed delivery of clotrimazole to the orbit. The dog retained sight in the affected eye, and clinical signs of infection were not detected 14 months later, although enophthalmos and medial strabismus may have been caused by persistent postinflammatory fibrosis. PMID- 10363097 TI - Fractures of the rostral portion of the mandible and maxilla in horses: 89 cases (1979-1997). AB - OBJECTIVE: To compare fracture locations, repair methods, complications, and outcomes of horses with fractures of the rostral portions of the mandible and maxilla. DESIGN: Retrospective study. ANIMALS: 89 horses with fractures of the rostral portions of the mandible and maxilla. PROCEDURE: Medical records and radiographs were reviewed. Fractures were categorized by fracture location and stability. Postoperative complications and long-term outcome were determined by clinical examination and telephone interviews with horse owners. RESULTS: 4 fracture types were recognized. Fractures involving just the alveolar plate (33%) and those involving the alveolar plate and the body of the bone (32%) were most common and were often repaired by interdental wiring. Unilateral fractures of the mandible (11%) were managed without surgery if stable. Unstable fractures were repaired with wires, a U-shaped bar (U-bar), or a bone plate. Bilateral fractures (24%) were often repaired with orthopedic wires in foals or with a U-bar, acrylic splint, wires, or bone plate in adult horses. In 2 horses, bilateral fractures were managed conservatively. Short-term complications developed in 24 of 89 (27%) horses. Soft tissue infections and wire loosening or failure were the most common short-term and long-term complications. Wire replacement was not required in any horses after release from hospital. Persistent draining tracts were most often associated with bone sequestration. Long-term functional and cosmetic outcomes were favorable for all fracture types and repair methods. CLINICAL IMPLICATIONS: Although complications in horses with fractures of the mandible and maxilla are common, long-term prognoses for functional and cosmetic outcome are favorable. PMID- 10363098 TI - Apical fracture of the proximal sesamoid bone in standardbred horses: 43 cases (1990-1996). AB - OBJECTIVE: To determine whether fracture fragment dimensions, suspensory ligament damage, and racing status at the time of injury were associated with outcome in Standardbred horses with apical fracture of the proximal sesamoid bone. DESIGN: Retrospective study. ANIMALS: 43 Standardbred racehorses. PROCEDURE: Medical records, racing records, and radiographs were reviewed, and ultrasonographic findings were scored. Measurements of the fractured portion of the proximal sesamoid bone were made. RESULTS: Seventy-four percent (32/43) of horses were pacers, and 26% (11/43) were trotters. Statistical differences between trotters and pacers regarding ability to start, number of starts, or amount of money earned after injury were not detected. Females earned significantly more money per start after injury than males. Eighty-six percent (37/43) of fractures involved hind limbs and 14% (6/43) involved forelimbs. Horses with forelimb injuries earned less money per start. Severity of suspensory ligament damage did not affect postinjury racing performance. A higher proportion of horses that had raced before injury returned to racing after surgery than horses that had not raced before injury, although a significant difference between these groups was not detected. Eighty-eight percent of horses that raced before injury raced after injury. Fifty-six percent of horses that did not race before injury raced after injury. Fracture fragment dimensions did not affect outcome. CLINICAL IMPLICATIONS: Dimensions of the apical fracture fragment of the proximal sesamoid bone in Standardbred horses and degree of suspensory ligament damage did not affect outcome. Prognosis for return to racing soundness is good in horses that had raced before injury and fair in horses that had not raced before injury. PMID- 10363099 TI - Nonsurgical treatment of suprascapular nerve injury in horses: 8 cases (1988 1998). AB - OBJECTIVE: To determine the outcome of horses with suprascapular nerve injury treated with stall rest alone. DESIGN: Retrospective case series. ANIMALS: 8 horses. PROCEDURE: Information on signalment, history, limbs affected, severity of lameness, degree of muscle atrophy, gait abnormalities, and results of radiography and electromyography was obtained from medical records. All horses were treated with stall rest. Follow-up information on severity of lameness, gait abnormalities, degree of muscle atrophy, time between injury and resolution of gait abnormalities, and outcome was obtained during reexamination at the hospital or through telephone conversations with owners. RESULTS: In 4 horses, the injury was a result of trauma; in the other 4, the injury was suspected to be a result of trauma. All horses had pronounced instability of the shoulder joint during the weight-bearing phase. Follow-up information was available for 7 horses. Shoulder joint instability resolved in all 7 horses within 3 to 12 months (mean, 7.4 months) after the original injury. Two horses had complete return of the supraspinatus and infraspinatus muscle mass 15 and 18 months after the injury. Two horses used as broodmares before the injury and 4 of 5 horses used for riding or in race training before the injury were able to return to preinjury activities. CLINICAL IMPLICATIONS: Horses with suprascapular nerve injury treated with stall rest alone have a good prognosis for recovery of normal gait and return to performance; however, the recovery period may be prolonged. PMID- 10363100 TI - Risk factors for abomasal displacement in dairy cows. AB - OBJECTIVE: To determine whether various periparturient events or 305-day milk production during the previous lactation period were associated with abomasal displacement in dairy cows. DESIGN: Retrospective, case-control study. ANIMALS: 75 pairs of case and control cows from 3 university-owned and 3 commercial dairy herds. PROCEDURE: Cows with abomasal displacement were matched with control cows on the basis of herd of origin, breed, age, and calving date. Frequency of specific periparturient events during the period from 2 weeks prior to parturition to diagnosis of abomasal displacement, as well as milk production during the preceding lactation period, were compared between case and control cows. RESULTS: Multivariate analyses indicated that case cows were significantly more likely to have had retained placenta, ketosis, a stillborn calf, metritis, twins, or parturient paresis than were control cows. Dystocia, mastitis, and milk production during the previous lactation period were not associated with abomasal displacement. CLINICAL IMPLICATIONS: Results indicated that a variety of periparturient events were associated with development of abomasal displacement among dairy cows. PMID- 10363101 TI - Associations among age, scrotal circumference, and proportion of morphologically normal spermatozoa in young beef bulls during an initial breeding soundness examination. AB - OBJECTIVE: To evaluate age and scrotal circumference as predictors of semen quality in young beef bulls. DESIGN: Cohort study. ANIMALS: 1,173 beef bulls < 15 months old. PROCEDURE: During initial breeding soundness examination, variables for bulls producing > or = 70% morphologically normal spermatozoa were compared with those for bulls producing < 70% morphologically normal spermatozoa. Mean and 95% confidence interval for age and scrotal circumference were constructed to detect differences between groups of bulls over all observations and within the 5 most common breeds. For these 5 breeds, chi 2 analysis was used to evaluate differences in the proportion of bulls that had values less than the population mean for scrotal circumference, age, and percentage of morphologically normal spermatozoa. Multivariate regression was used to quantify variation in the proportion of morphologically normal spermatozoa that could be explained by age and scrotal circumference. RESULTS: Mean (+/- SD) age and scrotal circumference differed significantly for bulls that produced > or = 70% morphologically normal spermatozoa (12.7 +/- 1.1 months and 35.6 +/- 2.7 cm) and bulls that produced < 70% morphologically normal spermatozoa (12.1 +/- 1.2 months and 34.8 +/- 3.3 cm). The proportion of bulls younger than mean age at first examination and the proportion producing > or = 70% morphologically normal spermatozoa differed among breeds. Age and scrotal circumference explained only 11% of the variation in semen quality. CLINICAL IMPLICATIONS: Among young beef bulls, those that were older and had larger testes were more likely to produce > or = 70% morphologically normal spermatozoa. Age and scrotal circumference were not sufficient predictors of semen quality. PMID- 10363102 TI - Use of a theloscopic triangulation technique for endoscopic treatment of teat obstructions in cows. AB - A method for endoscopic resection of obstructing tissue in the area of the teat canal opening into the teat sinus, using a triangulation technique, was developed. Benefits of this approach, compared with existing methods, include excellent observation of obstructing tissue, ease of manipulation of tissue flaps by use of a probe introduced through the teat canal, and precise excision of obstructing tissue by use of a pair of scissors introduced through an instrument portal. Outcomes for 12 cows treated with this technique were determined 2 weeks and 3 months after surgery. Three or more months after surgery, outcome was excellent in 10 of 12 cows, and 11 owners would opt for use of the procedure for other valuable cows in their herds. PMID- 10363103 TI - Ileocecocolic intussusception in a one-month-old llama. AB - A 1-month-old female llama was examined because of signs of acute abdominal pain. Physical and ultrasonographic examination revealed a cylindrical mass in the left caudal quadrant of the abdomen. Results of serum biochemical analyses and CBC were within reference ranges. Exploratory laparotomy revealed an ileocecocolic intussusception. Ileocecal resection and end-to-end ileocolic anastomosis were performed. After surgery, fecal examination revealed many coccidial oocysts, although a direct association between coccidiosis and intussusception could not be determined. The cria recovered without serious complications. PMID- 10363104 TI - Differences in morphology and cytoskeleton of MCF-7 and MX-1 cells after therapy with OH-tamoxifen and the pure estrogen antagonist ZM 182780. An immunofluorescence and scanning electron microscopic study. AB - The adjuvant endocrine therapy of breast cancer with non-steroidal antiestrogens of the triphenylethylene-type such as tamoxifen is clinically well established, and pure steroidal antiestrogens are being introduced in clinical trials to circumvent the probable occurrence of tamoxifen resistance. Nevertheless, there do still remain some unsolved questions about the exact mechanisms of these substances. We therefore investigated the different effects of 4-OH-Tamoxifen (OHT), a non-steroidal antiestrogen, versus ZM 182780, a pure steroidal antiestrogen, on the morphology and on the cytoskeleton of MCF-7 (estrogen receptor-positive) and MX-1 (estrogen receptor-negative) cells. For this purpose cells were treated for 2, 5 and 7 days with OHT, ZM182780 and different concentrations of beta-estradiol. Interestingly, in scanning electron microscopy, MCF-7 cells showed more differentiation by forming three-dimensional structures such as acini or tubule-like structures under ZM 182780 therapy than with OHT. As expected, MX-1 cells showed no effects after ZM 182780-therapy, but OHT led to a decrease in the number of these cells and produced a fibroblast-like appearance of the estrogen receptor-negative MX-1 cells. The following immunocytochemical experiments on the tubulin, vimentin, cytokeratin and actin cytoskeleton surprisingly did not show marked differences within the morphologically differentiated ZM 182780-treated population compared to the control group of MCF 7 cells. Only the OHT-treated cells of both, the ER(+) and the ER(-) cells, showed a rearrangement of actin filaments and cytokeratin which appeared even more pronounced within the ER(-) MX-1 cells. No experimental group showed morphologically detectable changes in tubulin or vimentin distribution. These data suggest a non ER-mediated OHT-effect on the cytoskeleton that also affects the ER(-) cell line MX-1. PMID- 10363105 TI - Scanning electron microscopy study including maceration techniques of an absorbable alloplastic implant designed for the reconstruction of ventral abdominal wall defects. AB - The interposition of synthetic material represents the most generally acknowledged method for achieving a tension-free surgical repair of major abdominal wall defects. As permanent materials are frequently associated with severe complications (rejection, peritonitis, enterocutaneous fistula, erosive invasion into the intestine), a newly designed absorbable prosthetic (polydioxanone, PDS) has been developed. The ellipsoid implant is composed of a knitted envelope and a filling of loosely arranged filaments. Interpositioning of the implant into an artificial circular abdominal wall defect was carried out on 30 Wistar rats. Explantation was performed 21 days post implantationem (p.i.), 42 days p.i. and 180 days p.i. The aim of the study was to evaluate the use of scanning electron microscopy (SEM) combined with maceration techniques (2 N NaOH for 1, 3 and 5 days) for a spatial assessment of the interactions between the implanted material and the ingrowing tissue components. Application of 2 N NaOH caused the complete dissolution of the PDS-material and a concomitant gradual disappearance of cellular and amorphous tissue components, thereby unmasking the remaining collageneous network. SEM of non-macerated specimens demonstrated that 21 days p.i. the entire implant has been filled with connective tissue components. Additionally, the ventral and dorsal surfaces of the implant were covered by a collageneous layer (neo-fascia). 21 days p.i. the PDS-filaments had developed minor clefts, which increased in number and depth 42 days p.i., and were transformed into small remnants 180 days p.i. Alkali treatment revealed the three-dimensional arrangement of collagen fibers, which ensheathed the PDS filaments and formed interconnecting networks between them. At the ventral portion of the implant the fibrous network was more elaborate and densely distributed. 180 days p.i. the implant has been transformed into a membranous structure (neo-membrane) composed of the ventral and dorsal neofascia, remnants of PDS-filaments and a continuous connective tissue layer containing wave-like collagenous structures. Whereas 21 and 42 days p.i. no herniation was observed, 130 days p.i. the implant began to bulge out of the ventral abdominal wall. It is therefore concluded that inspite of the advantages of absorbable materials, longer resorption times may be required to allow a sufficient consolidation of the ingrowing connective tissue to resist the tensile forces of the abdominal muscle coat. SEM combined with 2N NaOH maceration proved to be a usefull tool in addition to conventional histological techniques for a three-dimensional assessment of fibrous connective tissue components ingrowing into alloplastic implants. PMID- 10363106 TI - Germ cell migration and early development of the gonads in the trisomy 16 mouse- an animal model for Down's syndrome. AB - The aneuploid condition of patients with Down's syndrome (trisomy 21) frequently leads to a sub- or infertility of these individuals. Gonads from adults and fetuses with trisomy 21 demonstrated histologically a remarkable reduction in germ cells. Disorders in the germ cell migration, the early development of the gonads as well as meiotic defects are thought to contribute to this pathomorphology. To gain information about premeiotic defects, investigations on the trisomy 16 mouse, an animal model for Down's syndrome, were carried out. By means of morphometric studies a delay in migration and a reduction in primordial germ cells was evaluated in trisomic mice of embryonic day 11 (E11). At day E13 a generalized growth retardation of the developing gonads was obvious in trisomic animals. Additionally performed electron microscopic examinations revealed signs of germ cell demise in trisomy 16 mice. Thus, the mechanisms of a diminished proliferation capacity, impaired migration and premature death of germ cells represent premeiotic disorders that presumably contribute to the pathomorphology observed in the gonads of individuals with Down's syndrome. PMID- 10363107 TI - Scanning and transmission electron microscopic study of visceral and parietal peritoneal regions in the rat. AB - The visceral peritoneum of intraabdominal organs (spleen, stomach, liver, small intestine), omentum majus and the parietal peritoneum of the anterior abdominal wall and the diaphragm were studied in adult Wistar rats by combined scanning and transmission electron microscopy (SEM, TEM). In general, the peritoneal surface consisted of a mesothelium composed of cubic, flat or intermediate cell types delimited by a basal lamina. Cubic mesothelial cells predominated in parenchymal organs (spleen, liver) and were characterized by prominent and indentated nuclei, a cytoplasm richly supplied with organelles, a dense microvillous coat, basal invaginations and elaborate intercellular contacts. Flat mesothelial cells were observed in the intestinal, omental and parietal peritoneum (tendinous diaphragm, abdominal wall) and showed elongated nuclei, scant cytoplasm, a poorly developed organelle apparatus and sparsely distributed microvilli. An intermediate mesothelial cell type was described within the gastric peritoneum characterized by a central cytoplasmic protrusion at the nuclear region containing most of the cytoplasmic organelles and by thin finger-like cytoplasmic processes. The submesothelial connective tissue layer was composed of collagen fiber bundles, fibroblasts and free cells (macrophages, granulocytes, mast cells) and contained blood and lymphatic vessels. In the spleen, elastic fibers formed a membranous structure with intercalated smooth muscle cells. Mesothelial openings were observed as tunnel-like invaginations within the hepatic peritoneum and as clusters of peritoneal stomata within the parietal peritoneum of the anterior abdominal wall and the muscular diaphragm. The round or oval openings of the peritoneal stomata were frequently occluded by overlapping adjacent mesothelial cells and their microvillous coat or obstructed by cellular material. At the side of the peritoneal stomata the mesothelial cell layer was interrupted to allow a direct access to the underlying submesothelial lymphatic system. The mesothelium and lymphatic endothelium shared a common basal lamina. The endothelial cells were discontinuous and displayed valve-like plasmalemmatic interdigitations facilitating an intercellular transport of fluids and corpuscular elements from the peritoneal cavity to the submesothelial lymphatic lacunae. The findings underline the morphological heterogeneity of the peritoneum in visceral and parietal regions, suggesting different functional implications, and further support the presence of extra-diaphragmatic peritoneal stomata. PMID- 10363108 TI - Granin proteins (chromogranin A and secretogranin II C23-3 and C26-3) in the intestine of reptiles. AB - The occurrence, distribution and the possible cellular co-localizations of chromogranin A (CgA) and of two synthetic secretogranin II-peptides (SgIIC23-3 and SgIIC26-3) with several enteric neuropeptides and serotonin have been investigated immunohistochemically in turtles, lizards and snakes. The distribution of CgA-immunoreactivity was restricted only to the enteroendocrine cells in all the reptiles studied. SgII-immunoreactivity--absent in turtle- revealed nerve cells and fibers, besides enteroendocrine cells in lizard and snake guts. Moreover, the two antisera (C23-3 and C26-3) raised against the different regions of the SgII-molecule yielded distinct distribution patterns of immunoreactivity both in the lizard and snake organs. Small amounts of enteric serotonin cells co-stored CgA or SgIIC23-3 in lizards and snakes and only SgIIC26 3-peptide in snakes. CgA was found co-stored with somatostatin in a few enterocytes of the turtle duodenum. In the same gut segment of lizards and throughout the snake organ, neurotensin and the SgIIC23-3-peptide co-existed in a small number of endocrine cells. The pancreatic polypeptide-containing cells were devoid of immunoreactivity both for CgA and SgII. Bombesin immunopositive cells were absent throughout the intestines of the reptiles investigated. The above findings entirely support the heterogenous distribution of granins in neuroendocrine organs and tissues and also within the same neuroendocrine cell population. They further support the concept of a good conservation of granins during phylogeny. PMID- 10363109 TI - Detection of glycosidic residues in carpal glands of wild and domestic pig revealed by basic and lectin histochemistry. AB - Carpal glands are compound tubuloalveolar glands, located on the medial surface of the carpus. This study was carried out on samples from carpal glands of adult wild and domestic pigs of both sexes. We elucidated the glycosidic composition of carpal gland secretion in situ using traditional histochemical methods and lectin histochemistry. Some secretory cells exhibited an intense reaction with PAS in both wild and domestic pigs. Lectin histochemistry showed differences in the localization and composition of glycoconjugates secreted by carpal glands. A cytoplasmic positivity was revealed in the wild pig by the sequence sialidase-PNA and WGA, whereas in the domestic pig the reactivity was localized at the apical surface of some cells. LTA positive cells were found only in the carpal glands of the domestic pig. PMID- 10363110 TI - An alternative preparation of the acellular muscle graft for reconstruction of the injured nerve--morphological and morphometric analysis. AB - The application of cutaneous nerve grafts is accompanied by some disadvantages, including insufficient graft material for the reconstruction of a large mixed nerve. Recently, evacuated muscle autografts have been suggested as possible alternatives to cutaneous nerve grafts. In the present paper we have demonstrated a possible preparation of the evacuated muscle graft using an infiltration of Marcaine. The reinnervation of the distal stump of the rat median nerve was evaluated by morphological and morphometric analysis after application of the muscle acellular grafts prepared by three methods: an ordinary freeze-thawed muscle graft, a Marcaine evacuated muscle graft and a Marcaine treated graft with subsequent freezing and thawing. A comparison of the numbers and diameters of the myelinated axons in the distal nerve stumps revealed very similar conditions for axon regrowth and maturation in Marcaine evacuated and freeze-thawed muscle grafts. The best results with myelinated axon numbers and spectrum of their calibres were obtained when the Marcaine treated graft was repeatedly frozen and thawed. The pre-treatment of the muscle graft by Marcaine prevents it from shrinking and fragmenting, the main disadvantage during freeze-thawing of fresh muscle. The present results demonstrate that infiltration of striated muscles with a myotoxic compound, e.g. Marcaine, with subsequent freezing-thawing is the method of choice for the preparation of an acellular muscle graft used in peripheral nerve reconnection in the experimental model. PMID- 10363111 TI - [Histochemical and morphometric investigations of the musculature of forelimb mass of sheep with reference to its function. 1. Bending and extension of elbow joints]. AB - Muscle tissue was removed from the extensors and flexors of the elbow joint of six male sheep (180 days old) and stained for NADH tetrazolium oxidoreductase and myofibrillar ATPase after preincubation at pH 4.3 in order to identify three fiber types: slow twitch oxidative (STO), fast twitch oxidative (FTO) and fast twitch glycolytic fibers (FTG). The medial head of the M. triceps brachii and the anconaeus muscle had the largest fibers (> 50 and 60 microns). The smallest muscle fibers (35-43 microns) were found in the dorsal part of the long head of the triceps muscle. The medial head of the triceps muscle and the anconaeus muscle possessed a very high percentage of STO-fibers (90 and 100%) and FTG fibers were absent in these muscles. In the other extensors and flexors of the elbow joint the STO-percentage amounted to less than 30%. The dorsal part of the long head of the triceps muscle contained only 13% STO-fibers, but had the highest percentage of FTG-fibers (49%), which is representative of fast-muscles. The muscles of the elbow joint perform both static and dynamic functions. The medial head of the triceps brachii muscle and the anconaeus muscle possess the complement of enzymes which permits them to fulfil the work of extensors in the standing position. Therefore, they are typical of antigravity muscles. The histochemical structure of the other extensors and flexors reflect their function in motion. The lateral and long head of triceps muscle oppose the flexors and extend the elbow joint of the raised limb in the swing phase, during the landing phase they also function to support the other extensors of the elbow. PMID- 10363112 TI - Anatomy of the cranial nerve foramina in the Risso's dolphin (Grampus griseus). AB - In the present study, we examined the cranial nerve foramina of Risso's dolphin (Grampus griseus). There were two distinguishable characters in the cranial nerve foramina compared with terrestrial mammals. One was that the foramen infraorbitale was composed of several holes, but not a single hole. They should therefore be termed foramina infraorbitales. The infraorbital nerves ran through these foramina, went into the 'melon' and then branched in a complicated fashion. The facial nerve innervated the muscles surrounding the melon. A well-developed infraorbital and facial nerve complex may control the melon. Another was the presence of a porus acusticus internus and independent tympano-periotic bone. The separate ear bone forced the vestibulocochlear and facial nerves to exit from the cranial cavity through the porus acusticus internus. The independent ear bone structure may shut off the noise from the cranial bone to the periotic bone with a true receptor of hearing. It may be an adaptation for an acute sense of hearing. Compared with other dolphins, the cranial foramina of Risso's dolphin are definitely separate. The structure of the foramina is similar to that of pilot whales, but not to dolphins, so that Grampus may be closely allied to pilot whales. PMID- 10363113 TI - Shape of the orbital opening: individual characterization and analysis of variability in modern humans, Gorilla gorilla, and Pan troglodytes. AB - The description of the human orbital shape is principally qualitative in the classical literature, and characterised by adjectives such as circular, rectangular or quadrangular. In order to provide a precise quantification and interpretation of this shape, a study based on automatic image analysis and Fourier analysis was carried out on 45 human skulls (30 males, 15 females), and for comparison on 61 skulls of Gorilla gorilla (40 males, 21 females), and 34 skulls of Pan troglodytes (20 males, 14 females). Sexual dimorphism in the shape of the orbital opening was not demonstrated. Its dominant morphological features could be characterized by Fourier analysis; elliptical elongation and quadrangularity were dominant morphological features of the shape of the orbital opening in the three species. Elliptical elongation was more marked in humans and Pan, whereas quadrangularity was particularly emphasized in Gorilla. An intraspecific variability of the shape of the orbital opening existed in humans, Gorilla and Pan, and seemed close in the three species. Interspecific partition between humans, Gorilla and Pan was demonstrated despite the variability observed in the three species studied. Interspecific differences between Gorilla and the Pan-humans group were principally explained by the differences in quadrangularity, and by differences in orientation of triangularity and pentagonality. Differences in the shape of the orbital opening between humans and Pan were principally explained by differences in hexagonality, and by differences in orientation of quadrangularity. A closeness of shape between some humans and some individuals in Pan and, to a lesser degree, with some individuals in Gorilla was observed, demonstrating the existence of a morphological continuum of the shape of the orbital opening in hominoids. PMID- 10363114 TI - ["One must always be true to himself." The proper ethical reservation on so called art anatomy]. PMID- 10363115 TI - Quality assessment of computer-assisted semen analysis (CASA) in the andrology laboratory. AB - If quality is assessed with regard to computer-assisted semen analysis (CASA), the evaluation of seminal fluid in the andrological laboratory has to be considered. Three levels of quality assessment are generally accepted: structure, process and results. Quality of structure mainly concerns the quality of laboratory assessment, in particular the skill of the staff and the equipment used. The quality of the CASA system itself is difficult to assess. Process quality concerns the quality of performing a diagnosis. When the parameter settings of the CASA system and the handling of the sample are defined, the reproducibility of the CASA values is clearly better than that of the visual estimation of motility. CASA systems are also superior to other methods regarding the documentation of laboratory values, as all the values are obtained directly online. Result quality comprises the precision, reliability and reproducibility of measurement as well as the significance of values with respect to their biological relevance. Concluding from the definitions as quoted above and from reports of the literature it may be stated that: (i) in the dimensions of structure and process quality, CASA is superior to other methods of measuring sperm motility; (ii) the evaluation of results and quality of results, however, is highly problematic; (iii) CASA systems do not appear to be superior to the visual estimation of sperm motility with respect to the fertilizing capacity of spermatozoa; (iv) the guidelines of the WHO task force form a basis for sufficient process quality; (v) further efforts should actually focus not on the improvement of investigation technology, but on the improvement in the qualification of investigators. PMID- 10363116 TI - Sperm penetration through the human zona pellucida as a predictor of in vitro fertilization. AB - The aim of this study was to determine the predictive value of sperm penetration into the perivitelline space of human cadaveric oocytes on in vitro fertilization outcome. Forty-two patients with tubal infertility undergoing ovarian stimulation with gonadotropin for in vitro fertilization and embryo transfer participated in the study. The number of spermatozoa bound to the human zona pellucida, the percentage of cadaveric oocytes with one or more spermatozoa in the perivitelline space, and the in vitro fertilization outcome were evaluated. Spermatozoa from 37 of 42 patients were able to penetrate the perivitelline space of cadaveric oocytes as well as to fertilize human oocytes in vitro. In three individuals, no penetration of the perivitelline space of cadaveric oocytes was observed and no in vitro fertilization was detected. Only two patients were able to fertilize the couple's oocytes without penetration of the cadaveric oocytes. Based on these results the specificity and the sensitivity of the assay to predict in vitro fertilization was 100% and 94.1%, respectively. Accordingly, these results suggest that sperm-zona penetration is a useful bioassay to predict male fertility potential in IVF outcome. PMID- 10363117 TI - Serum levels of inhibin B in men with different causes of spermatogenic failure. AB - Inhibin B appears to be the physiological feedback signal for FSH. Herein the determination of serum levels of inhibin B, FSH, LH and testosterone in 148 infertile patients and their association with clinical findings and seminal parameters are reported. A significant negative correlation of FSH and inhibin B (r = -0.60) was found. LH levels showed a significant inverse correlation (r = 0.37), but a weak regression (c0 = 0.01). No correlation with testosterone levels occurred. A significantly positive correlation was observed between testis volume and inhibin levels (r = 0.39) as well as between sperm count and inhibin levels (r = 0.39). To evaluate whether the secretion of inhibin B depends on the nature of damage to the Sertoli cells, inhibin levels in 23 patients with varicocele; eight after cryptorchidism, and 16 after hemiorchiectomy were compared to those of other patients without these diseases, but identical sperm count. No significant differences were found. In 20 men undergoing testicular biopsy, inhibin levels were compared to histology. Although the men with Sertoli-cell only syndrome had significantly lower levels ((15.83 +/- 12.2) pg ml-1) than those with normal spermatogenesis ((183.8 +/- 112.3) pg ml-1), a distinction between patients with hypospermatogenesis from those with normal spermatogenesis was not possible on the basis of inhibin levels. Between these groups, the distinction was better achieved by the FSH levels (sensitivity of 85%). We conclude that inhibin B levels are a serum marker of Sertoli cell function, but the prediction of the quality of spermatogenesis is not superior to that of FSH levels. PMID- 10363118 TI - Sex differences and similarities of hormonal alterations in patients with anorexia nervosa. AB - The aim of the present study was to gain better insight into hormonal disturbances in male patients with anorexia nervosa. It included six men with anorexia nervosa aged 13-26 years, with a mean body weight of 42.83 +/- 8.03 kg, a body mass index of 15.08 +/- 1.26 and an average degree of weight loss 29.98 +/ 4.73%. The results were compared with those of 15 healthy age-matched males and 40 women with anorexia nervosa. Prolactin, growth hormone and the gonadal and thyroid axis were studied in detail. The gonadotropin basal levels and their responses to gonadotropin-releasing hormone in male patients were lower, but not significantly, in comparison with healthy men. The basal levels and the responses of luteinizing hormone in anorexic women were significantly lower in comparison with female controls, but the decreased basal level of follicle-stimulating hormone showed an exaggerated response to gonadotropin-releasing hormone. In male anorexics the testosterone levels (7.1 +/- 10.9 nmol l-1) were significantly lower. The changes in the thyroid axis and in prolactin secretion were almost the same in male and female patients. The data of this study suggest that endocrine disturbances in males are similar to those in females with anorexia nervosa, but differences exist mainly in relation to the gonadal axis. PMID- 10363119 TI - Different cumulative pregnancy rates in patients with repeated IVF- or ICSI cycles: possible influence of a male factor. AB - The low rate of ongoing pregnancies in IVF cycles leads to a high number of repeated cycles in couples with previously failed attempts. Therefore it would be helpful to have a prediction about the chance of becoming pregnant in a repeated cycle. In a retrospective study the data of about 4246 cycles were analysed. Because the pregnancy rates in IVF- and ICSI cycles are generally different, these two groups were distinguished between and the outcome in patients with one, two or more attempts was analysed. The rate of ongoing pregnancies per patient was lower after IVF (24.9%) than after ICSI (32.9%), but was similar or even slightly increased in patients with more than one attempt. On the other hand, there was a high pregnancy rate with ICSI in the first two cycles (35.9%), but patients with more than two ICSI cycles had a significantly lower chance of becoming pregnant (20.7%). Factors that are known to influence the pregnancy rate, such as stimulation protocol, oocyte quality or number of transferred embryos, were similar in all groups. However, significantly reduced embryo quality with successive cycles was only observed in ICSI patients. There might be a negative selection of patients with poor embryo quality and previously failed attempts after ICSI, possibly due to an andrological factor. The differences between IVF- and ICSI patients are based on treatment indications, and andrological diseases are the predominant indication for ICSI. Although no correlation was found between changes in conventional sperm parameters and number of treated cycles, there might be a subgroup of andrological patients selected by repeatedly failed ICSI cycles. Chromosomal or genetic disturbances in spermatozoa of this subgroup could be the reason for failure. PMID- 10363120 TI - Isolation method of Leydig cells from mature male Djungarian hamsters (Phodopus sungorus) and their steroidogenic activity in vitro. AB - For studies addressing the functions of Leydig cells, isolated cells are often better suited than intact animals. Here, the isolation procedure of Leydig cells from adult male Djungarian hamsters (Phodopus sungorus) is described. Cells were isolated using a procedure involving enzymatic dissociation and Percoll-gradient centrifugation. For each experiment, approximately 4.4 x 10(6) Leydig cells from six animals were obtained. The cells showed high steroidogenic responsiveness to physiological (ovine luteinizing hormone (oLH) and human chorionic gonadotropin (hCG)) and nonphysiological (forskolin) stimuli in vitro. Approximately 98% of cells were viable as assessed by trypan blue exclusion, and the purity varied from 80 to 95% as tested by 3 beta-hydroxysteroid dehydrogenase activity. Leydig cells were also identified by a bright yellow halo under phase-contrast microscopy. They contained numerous lipid droplets and showed round nuclei and prominent nucleoli. The cells responded to oLH, hCG and forskolin with an increased testosterone production in a dose-dependent manner. Dose-response curves in these studies suggest that Leydig cells of Djungarian hamsters undergo desensitization, probably due to down regulation of their LH/CG receptors. PMID- 10363121 TI - Repeatability and variance analysis on multiple computer-assisted (IVOS) sperm morphology readings. AB - The repeatability of the Hamilton Thorne Research IVOS (version 10) semen analyser (dimension specific software, version 3) in the evaluation of sperm morphology according to strict criteria was investigated in this study. The repeat measures investigated were cell-cell (300 cells, 3 x each), intraslide (20 slides, 3 x each) and interslide (30 samples, 3 slides each), and their normal sperm morphology outcomes were recorded. Semen samples with varying normal sperm morphology percentages were obtained and sperm morphology slides prepared. The slides were stained with Diff-Quik stain. Agreements between evaluations were determined using the kappa statistic and average coefficients of variation. The predictive probability for an abnormal cell given a prior abnormal cell outcome was 91%, and 89% for a similar prediction of a normal cell. The predictive probabilities for an abnormal or a normal cell given two prior abnormal or two prior normal cell outcomes were 95% and 94%, respectively. No significant bias was obtained between the repeat probabilities for normal and abnormal sperm cells. The average coefficients of variation for the intraslide trial were 9.73% and 8.30% when 100 and 200 sperm cells were evaluated, respectively. The average coefficient of variation for the interslide trial was 15.39%. The technical importance of good sample and slide preparation technique has once again been highlighted by this study. A uniform (spatial homogeneity), high concentration (5 10 cells per computer screen) smear must be made and the cells stained with optimal intensity (maximum contrast). In a trial in which 2000 cells were evaluated, 19 objects (0.95%) were identified as spermatozoa, but were debris. The automated semen analysing system (IVOS) used in this study was shown to maintain a level of repeatability, precision and accuracy acceptable for the application of the system in a routine semen analysis situation. PMID- 10363122 TI - Ongoing pregnancies resulting from intracytoplasmic sperm injection (ICSI) of spermatozoa from frozen-thawed testicular biopsy specimens. AB - Two clinical pregnancies following intracytoplasmic sperm injection of spermatozoa from frozen-thawed testicular biopsies in two azoospermic men are reported. The use of spermatozoa from cryopreserved testicular tissue is therefore a viable option for azoospermic men, as our results indicate that pregnancies is achievable in these cases. PMID- 10363123 TI - Does transcervical intra-fallopian insemination improve pregnancy rates in cases of oligoteratoasthenozoospermia? A prospective, randomized study. AB - The relatively low pregnancy rates (PR) after treatment of patients with oligoteratoasthenozoospermia (OTA) result in a search for different treatment modalities. The objective of this study was to assess the efficacy of transcervical intrafallopian insemination (IFI) with husband's semen in comparison to intrauterine insemination (IUI) in couples with OTA. A prospective, randomized study included 30 couples with OTA-related infertility (according to WHO criteria). The female patients underwent individually adjusted controlled ovarian stimulation by gonadotropins. Spermatozoa was prepared using the Percoll 70% technique and insemination was performed 36-40 h after human chorionic gonadotropin (HCG) administration. The Tomcat Catheter was used for IUI and the Jansen-Anderson Catheter for IFI to the fallopian tube leading to the ovary that contained more dominant follicles. The couples were divided according to sperm count, into group A (9 couples): < 10 mill ml-1 and group B (21 couples): > 10 mill ml-1. Within the groups the patients were randomly assigned for IUI or IFI treatment. Among group B couples, two pregnancies out of 15 IUI cycles (13.3% PR) and two pregnancies out of 18 IFI cycles (11.1% PR) were achieved. Group A patients completed 7 IUI and 9 IFI treatment cycles with no pregnancies observed. These data did not demonstrate a statistically significant advantage for either technique. PMID- 10363124 TI - Recovery of ejaculated spermatozoa for intracytoplasmic sperm injection after anti-inflammatory treatment of an azoospermic patient with genital tract infection: a case report. AB - In this paper our experiences with anti-inflammatory treatment of an infertile patient with azoospermia and concomitant silent genital infection are reported. The patient was referred to our fertility centre with prediagnosed non obstructive azoospermia and no spermatozoa were found in the ejaculate on two occasions. The patient showed leukocytospermia and was suspected to be affected by genital infection. Therefore, anti-inflammatory treatment was initiated and 8 weeks later examination of the ejaculate revealed a decreased number of leukocytes and the presence of few but motile spermatozoa. Subsequently, two ICSI cycles were performed with anti-inflammatory therapy in parallel and a sufficient number of spermatozoa could be retrieved for injection. However, in a third cycle without previous treatment, examination of the ejaculate again revealed azoospermia and leukocytospermia. It is concluded that, in cases of azoospermia and chronic genital infection, some patients will benefit from anti-inflammatory treatment prior to and during ICSI treatment. This may allow retrieval of spermatozoa from the ejaculate and thus may avoid the need for a therapeutic testicular biopsy. Using this approach, sufficient spermatozoa in three out of four ICSI cycles could be retrieved and a pregnancy was finally achieved. PMID- 10363125 TI - Human aneuploidy: lessons from achiasmate segregation in Drosophila melanogaster. AB - Aneuploidy is a crucial issue in human reproductive biology, accounting for both a significant proportion of miscarriages and, among liveborns, multiple congenital malformation syndromes such as Down Syndrome. Although the etiology of human aneuploidy remains poorly understood, recent studies have elucidated certain fundamental correlates of meiotic nondisjunction, such as altered recombination. These features are extraordinarily similar to those associated with chromosome misbehavior in Drosophila melanogaster females. Furthermore, these organisms also share a significant level of achiasmate chromosome nondisjunction. Here we describe in detail the processes of achiasmate chromosome segregation in Drosophila and discuss how they may be most effectively applied to our understanding of the etiology of human aneuploidy. In particular, we examine the possibility that similar "backup" mechanisms of chromosome segregation might function in mammalian meiosis, particularly mammalian females. Drawing upon observations made in flies, we also propose a new model for the segregation of achiasmate chromosomes in humans. PMID- 10363126 TI - Linkage disequilibrium at the cystathionine beta synthase (CBS) locus and the association between genetic variation at the CBS locus and plasma levels of homocysteine. The Ears II Group. European Atherosclerosis Research Study. AB - Cystathionine beta synthase (CBS) is a key enzyme in homocysteine metabolism. We have examined four apparently non-functional polymorphisms in the CBS gene and have determined their frequency, degree of linkage disequilibrium and association with plasma homocysteine levels. The polymorphisms are a 68 bp insertion in exon 8, C699T in exon 8, C1080T in exon 11 and C1985T in the 3' untranslated region. 785 individuals participating in the European Atherosclerosis Research Study II (EARSII), from 11 countries across Europe were genotyped for these polymorphisms. The 68bp insertion had the highest frequency in the UK and in the Middle region, with a lower frequency in the Baltic and the South (p = 0.01), and the exon 11 polymorphism had the highest frequencies of the rare allele in the Baltic (p < 0.05). There was a high degree of linkage disequilibrium between the polymorphisms (p < 0.001 overall), except between C699T and the C1985T, with three common haplotypes accounting for nearly 80% of chromosomes. Examination of the association between these polymorphisms and plasma homocysteine levels revealed that the carriers of the rare alleles of the C699T, C1080T and C1985T polymorphisms had lower plasma homocysteine concentrations than those homozygous for the common alleles, although these differences were not statistically significant. The thermolabile valine variant caused by a substitution of a C for a T at nucleotide 677 in the methylenetetrahydrofolate reductase (MTHFR) has previously been shown to have profound effects on plasma levels of homocysteine in this sample, but the homocysteine-raising effect associated with this thermolabile variant was not seen in carriers of the 68 bp insertion, with this interaction being statistically significant (p < 0.001). These data demonstrate that variation in the CBS gene as detected with these four polymorphisms, had no statistically significant effect on plasma homocysteine levels in these healthy young men. However, the presence of the 68 bp insertion, which is found in approximately 7.5% of individuals in the populations of Europe sampled, abolishes the raising effect of thermolabile MTHFR Val/Val genotype, and may be of importance in the situation of high homocysteine. PMID- 10363128 TI - Segregation analysis of squamous cell carcinoma of the head and neck: evidence for a major gene determining risk. AB - We and others have shown that a family history of squamous cell carcinoma of the head and neck (SCCHN) is a risk factor for this disease. We performed a segregation analysis on a dataset comprised of 1429 first-degree relatives of 242 unselected cases of SCCHN and 934 first-degree relatives of 156 spouse controls. Using the SAGE software, we demonstrated that a Mendelian model was favored and a model postulating a purely environmental cause of SCCHN was rejected. The model suggests that 18% of the population who smoke and drink are susceptible. The lifetime risk for non-smokers and non-drinkers who are heterozygotes for the susceptible allele is close to zero, but for those heterozygotes who smoke and drink the risk is 14% by age 70. These findings suggest that specific genetic factors account for a significant fraction of the risk of SCCHN associated with a family history of SCCHN. PMID- 10363127 TI - Comprehensive mutation analysis of TSC1 using two-dimensional DNA electrophoresis with DGGE. AB - Tuberous sclerosis complex (TSC) is an autosomal dominant disorder characterised by the development of benign tumors in multiple organs often causing serious neurologic impairment. To develop a reliable genetic test for TSC, two dimensional electrophoresis with denaturing gradient gel electrophoresis (2D DGGE) has been developed to detect mutations in TSC1. The 23 exons of TSC1 were amplified using two rounds of PCR. In the first round, all coding regions of TSC1 were amplified in four fragments ranging in size from 7.4 kb to 9.9 kb. In the second round, 32 fragments representing 23 exons were amplified using primers designed to avoid overlapping fragments and with a GC clamp on one end to optimise melting characteristics. These exon fragments were then separated by size in the first dimension using a polyacrylamide gel, and by melting characteristics in the second dimension using a urea/formamide gradient to yield 32 distinct bands. If a mutation is present, four bands instead of one, are typically observed. During the development of this assay, we analysed 63 patient samples with known TSC1 mutations from prior studies. These 63 patients had 68 known mutations or polymorphisms. With DGGE, all 68 of these were identified (45 point mutations, 3 small insertions, 20 small deletions) and an additional 27 single base variants were discovered. To evaluate the assay, we analysed 19 of these samples in a blinded study. In the blinded analysis, 19/20 (95%) known mutations or polymorphisms were detected. The single missed mutation in the blinded analysis could be identified in retrospect and the assay was modified accordingly. During this study, we identified 2 new mutations (exon 8 and exon 15), a new polymorphism (intron 4), and the first variant identified in a non coding exon (exon 2). PMID- 10363129 TI - Carrier detection by microsatellite analysis of Duchenne/Becker muscular dystrophy in Hungarian families. AB - Duchenne and Becker muscular dystrophies are among the most severe and frequent inherited disorders. Being still incurable, medical treatment is concentrated on the carrier diagnosis of the members of the affected families. Here we report the results of the studies of 151 members of 41 Hungarian families, obtained with multiplex PCR amplification of 18 exons as well as the muscle specific promoter region, and haplotype analysis of two polymorphic (CA)n repeat microsatellite loci in introns 45 and 49 of the dystrophin gene. The analysis of 15 deletion type families revealed a frequency of new mutations not differing significantly from that in the other regions of Europe. We also compared the allele distributions of the two microsatellites in randomly selected normal individuals and affected family members. The allele distribution of STRP45 shows interesting differences between the two populations. PMID- 10363130 TI - Genetic linkage analysis using lognormal variance components. AB - Typically genetic studies of continuous traits such as cholesterol levels or blood pressure assume that interindividual variability follows a normal distribution. Here we develop methods to analyze positively skewed data by assuming a lognormal distribution. We develop a variance components approach for identifying such effects from a major gene, residual polygenic factors and nongenetic factors. We compare by a simulation study results from fitting this lognormal model with either applying the log transformation or not transforming the data. We found that the lognormal model provided more precise estimates and more powerful tests than a simple log transformation when analyzing lognormally distributed data. Power varied with sibship size. For the same total number of nonindependent sibpairs, larger sibships were less powerful. However, larger sibships are more economical because they require a smaller sample size to obtain a specified power. To illustrate the application of this lognormal model to real data, we studied evidence for linkage between triglycerides and the lipoprotein lipase gene. PMID- 10363131 TI - Mitochondrial DNA analysis of northwest African populations reveals genetic exchanges with European, near-eastern, and sub-Saharan populations. AB - Genetic studies have emphasized the contrast between North African and sub Saharan populations, but the particular affinities of the North African mtDNA pool to that of Europe, the Near East, and sub-Saharan Africa have not previously been investigated. We have analysed 268 mtDNA control-region sequences from various Northwest African populations including several Senegalese groups and compared these with the mtDNA database. We have identified a few mitochondrial motifs that are geographically specific and likely predate the distribution and diversification of modern language families in North and West Africa. A certain mtDNA motif (16172C, 16219G), previously found in Algerian Berbers at high frequency, is apparently omnipresent in Northwest Africa and may reflect regional continuity of more than 20,000 years. The majority of the maternal ancestors of the Berbers must have come from Europe and the Near East since the Neolithic. The Mauritanians and West-Saharans, in contrast, bear substantial though not dominant mtDNA affinity with sub-Saharans. PMID- 10363132 TI - Assignment of the human ARNO3 gene (PSCD3) to chromosome 7p21 by radiation hybrid mapping. AB - The chromosomal localization of the human ARNO3 gene (PSCD3) was determined using monochromosomal somatic cell hybrid and radiation hybrid mapping panel. PCR analysis of a hybrid DNA panel localized the ARNO3 gene to human chromosome 7. The analysis of the Genebridge4 radiation hybrid panel using PCR amplification located the ARNO3 gene between NIB1822 (9.5 cR) and D7S481 (5.5 cR) with a lod score of > 3.0. This region is located in the human chromosome band 7p21. PMID- 10363133 TI - The biology of breast cancer. AB - This article focuses on the major hormones and growth factors for which a critical role in normal mammary growth has been clearly defined. Certainly other hormonal systems and growth factors could also affect breast cancer initiation and progression, but their exact contribution to normal and/or malignant breast cell growth is poorly delineated. Examples of such factors include somatostatin, mammostatin, mammary-derived growth inhibitor (MDGI), mammary-derived growth factor-1 (MDGF-1), inhibins, activins, androgens, glucocorticoids, vitamin D, thyroid hormones, ecosinoids, and oxytocin. Clearly, the hormonal regulation of breast cancer cell growth and survival is multifaceted and very complex. In particular, the effects of estrogens and anti-estrogens on breast cells may depend on their interaction with a wide variety of other pathways. In addition, these interactions may vary among individual breast tumors depending on other genetic changes in the tumor cells that have not been discussed here, such as oncogene activation and loss of tumor suppressors. A more detailed understanding of how cells circumvent a dependency on these pathways is greatly needed in order to identify new biological targets and to design novel therapies for breast cancers that are resistant to anti-estrogen therapy. Such agents could be used alone or in combination with anti-estrogens to improve response to a second course of hormonal therapy. PMID- 10363134 TI - Advances in breast imaging. AB - Many exciting developments are occurring in breast imaging. Digital mammography holds the promise of telemammography and computer-aided diagnosis. Mammoscintigraphy may be helpful in identifying drug-resistant tumors before therapy. There is renewed interest in evaluating ultrasound as a potential adjunctive screening tool in women with radiographically dense breasts. Finally, contrast-enhanced magnetic resonance imaging may be used more extensively in the monitoring of tumor response to primary chemotherapy and in the preoperative workup of patients being considered for breast conservation therapy. PMID- 10363135 TI - Lymphatic mapping in breast cancer. AB - The most accurate predictor of survival in breast cancer is the presence or absence of lymph node metastases. Lymphatic mapping with sentinel node biopsy is a new technique that provides more accurate nodal staging compared with routine histology for women with breast cancer, but without the morbidity of a complete lymph node dissection. Sentinel lymph node (SLN) biopsy is a more conservative approach to the axilla that requires close collaboration from the surgical team, nuclear medicine, and pathology. National trials are investigating the clinical relevance of the upstaging that occurs with a more intense examination of the SLN. As is the case with breast preservation as a viable alternative to mastectomy for the definitive treatment of the primary node, selective lymphadenectomy has the ability to decrease morbidity without compromising patient care. PMID- 10363136 TI - An update on radiation therapy in breast cancer. AB - Radiation therapy plays an important role in the management of both invasive and noninvasive breast cancer. During the last 20 years, the availability of radiation therapy has made it possible to test the feasibility and safety of breast preservation after the diagnosis of early-stage breast cancer. This article summarizes some of the ongoing controversies concerning the use of radiation therapy in the multidisciplinary management of breast cancer. PMID- 10363137 TI - Adjuvant therapy for resectable breast cancer. AB - The breast cancer mortality rate is falling, most likely because of a combination of early detection, refined surgical and radiation therapy techniques, and improved systemic therapy efficacy. The proper integration and application of these treatment modalities present evolving challenges for clinicians. Systemic therapy, in particular, is changing rapidly with the advent of new chemotherapy drugs, new classes of agents, and new therapeutic regimens. The most recent studies suggest that optimal outcomes are possible through the broad but appropriate use of hormone therapy and chemotherapy to prevent relapse and possibly prevent second primary tumors. The choice of therapy for patients remains a matter for careful consideration and discussion in each individual case. PMID- 10363138 TI - The role of chemotherapy for metastatic breast cancer. AB - Metastatic breast cancer remains a devastating and largely incurable disease. Currently available therapies offer meaningful palliation for many and modest prolongation of survival for some patients. Single-agent hormonal therapy remains the treatment of choice for patients with ER-positive disease, with sequential use of further hormonal agents or cytotoxic chemotherapy at the time of disease progression. Chemotherapy is appropriate as initial therapy for patients with receptor-negative or rapidly progressive visceral disease. Although combination regimens may increase response rates, the lack of survival benefit does not justify the increased toxicity of aggressive combination regimens in most patients. Maintenance chemotherapy deserves consideration in selected well informed patients, especially those with few therapy-related side effects. High dose regimens confer substantial toxicity with no clear therapeutic advantage and cannot be recommended outside of ongoing trials. New chemotherapy agents offer the hope of effective salvage therapy with acceptable toxicity to a larger number of patients. Perhaps most promising, the development of targeted, biologically based therapies such as rhuMAbHER2 offers encouragement for the future. PMID- 10363139 TI - Hormonal therapy for breast cancer. An update. AB - The use of endocrine manipulation for the treatment of breast cancer has been available for 100 years, but in recent years the number of therapeutic options available to patients has increased dramatically. This article considers new developments in the use of hormonal agents for the treatment and prevention of breast cancer. PMID- 10363140 TI - Locally advanced breast cancer. AB - The multidisciplinary management of patients with locally advanced breast cancer (LABC) has evolved over the last three decades. The introduction of chemotherapy (CT) and endocrine therapy in conjunction with surgery and radiation therapy (XRT) have changed the natural history of this disease. The authors discuss the role of the taxanes, novel endocrine therapy, and high-dose CT programs in the management of LABC. Indications for breast-conserving surgery are presented. The role of prognostic and predictive tumor markers in LABC is discussed. PMID- 10363141 TI - [Measurements and clinical usefulness of carboxyterminal propeptide of type I collagen and cross-linked carboxyterminal telopeptide of type I collagen in patients with prostate cancer]. AB - Carboxyterminal propeptide of type I collagen (P1CP) and cross-linked carboxyterminal telopeptide of type I collagen (1CTP) are known as parameters of bone metastasis in patients with prostate cancer (PCa). We measured the serum P1CP and 1CTP levels in 52 PCa patients and evaluated the clinical usefulness of these serum markers. Both serum levels of P1CP and 1CTP were significantly higher in patients with extent of disease (EOD) grade 2 or 3 bone metastases than in patients without bone metastasis. Thus, P1CP and 1CTP are not as useful at first detection of bone metastases as bone scintigram. On the other hand, in the patients who indicated high serum levels of P1CP or 1CTP before initial treatment, the changes in the concentrations of these markers may be helpful in evaluating the response to treatment or the progression of disease. Our results suggest that P1CP and 1CTP are useful markers for monitoring the metastatic burden in the bone of PCa patients, but the efficacy is limited in high EOD grade cases. PMID- 10363142 TI - [Transurethral resection of the prostate for patients with dementia]. AB - During the period from July 1995 to June 1996 we performed transurethral resection of the prostate (TURP) on 824 patients with benign prostatic hyperplasia (BPH). Among them, 13 were dementia patients between 74 and 96 years old; they presented with urinary hesitancy in 6, retention in 4, frequency in 2 and incontinence in 1 patient. Past history included stroke in 7, hypertension in 6, pulmonary tuberculosis in 4, diabetes in 3, asthma in 2, angina pectoris in 1, Parkinson's disease in 1, pneumonia in 1, and hepatitis in 1. Careful preoperative examination revealed that they were proper candidates for TURP. They underwent TURP under spinal anesthesia. The mean operative time was 34 min, ranging from 20 to 60 min. The adenoma resected weighed 24 g on the average, ranging from 7.5 to 48 g. During surgery, although hypotension was noted in 2 patients, there was no serious morbidity. Their mental condition was well controlled with ketamine and diazepam during and after surgery. Postoperative complications included acute myocardial infarction in 1, multiple gastric ulcer in 1, and decubitus in 1. None died within 3 months after TURP, 3 died there after, and 10 patients were alive at the mean follow-up period of 26 months. Six patients reported good urination, 3 reported some improvement in urination after surgery, although requiring intermittent catheterization and 1 developed mild incontinence. In conclusion, TURP appears to provide some benefit in selected patients with dementia and should not be considered to be a contraindication for such patients. PMID- 10363143 TI - [ACTH-independent bilateral macronodular adrenocortical hyperplasia (AIMAH): a case report]. AB - We report a case of adrenocorticotropic hormone (ACTH)-independent macronodular adrenocortical hyperplasia (AIMAH). A 54-year-old Japanese man was admitted to our hospital for further examination of obesity and hypertension. Endocrinological studies showed that plasma cortisol was high (22.5 micrograms/dl) without diurnal rhythm, and plasma ACTH was low. Two or 8 mg of dexamethasone did not suppress the plasma cortisol levels. Abdominal computed tomography revealed nodular hyperplasia of bilateral adrenal glands. Adrenal scintigraphy showed the positive uptake of 131I-adosterol to bilateral adrenal glands. Brain magnetic resonance imaging revealed no abnormalities. He was diagnosed as having Cushing's syndrome with bilateral adrenal hyperplasia, and bilateral adrenalectomy was performed. Left and right adrenal glands were 52 g and 35 g, respectively, and were occupied by yellow nodular lesions. Histologically, hyperplastic lesions were composed of clear cells. Finally he was diagnosed with AIMAH. PMID- 10363144 TI - [A case of idiopathic retroperitoneal fibrosis with renal subcapsular urinoma resolved by steroid therapy]. AB - A case of idiopathic retroperitoneal fibrosis is reported. The patient was a 63 year-old man with the complaint of right flank pain, general fatigue and weight loss. Intravenous pyelography revealed right hydronephrosis and peripelvic extravasation. Abdominal computed tomography showed subcapsular urinoma of the right kidney and mass lesion surrounding the aortic bifurcation. Retrograde pyelography demonstrated a narrow segment at the middle portion of the right ureter through which the ureteral catheter could be easily passed. Magnetic resonance imaging showed a low-intensity mass on T1 weighted images and a heterogeneous mass on T2 weighted images. Steroid therapy was selected under the diagnosis of idiopathic retroperitoneal fibrosis with subcapsular urinoma of the right kidney. Prednisolone was administered for 60 days, resulting in the complete disappearance of the urinoma. PMID- 10363145 TI - [Spontaneous rupture of renal cell carcinoma: a case report]. AB - A 74-year-old man with severe right flank pain and hypochondralgia, was admitted to a hospital where he was found to have an abnormality of the right kidney on computed tomographic (CT) scan. He was referred to our department for further examination and treatment on the next day. Spontaneous rupture of the right renal cell carcinoma was mostly suspected from preoperative clinical findings obtained by ultrasonography. CT scan and angiography. Extravasation was not recognized on angiography. We chose emergent transcatheter arterial embolization prior to radical nephrectomy. The surgical specimen contained a solid and yellowish mass invading into the renal pelvis. Subcapsular rupture was identified. Histopathological diagnosis was renal cell carcinoma consisting of invasive growth of highly atypical epithelial cells with a sarcomatous pattern, and the tumor cells were present in the renal pelvis. He died of lung cancer 26 months after the operation. PMID- 10363146 TI - [A case of synchronous multifocal urothelial tumors in a patient with phenacetin abuse]. AB - A 79-year-old male with phenacetin abuse was admitted to our University Hospital for treatment of asymptomatic gross hematuria. Intravenous urograpdy and computed tomography revealed synchronous right renal pelvic carcinoma and bladder carcinoma. Right nephroureterectomy and transurethral resection of bladder tumor (TUR-Bt) were performed. Histologically, right renal pelvic tumor and bladder tumor were both transitional cell carcinomas of grade 2, pT1, and grade 1 = 2, Ta, respectively. Additionally, pathological examination revealed two distal ureteral tumors, which were transitional cell carcinomas of grade 2, pTa. He also had a history of heavy tobacco-smoking (20 cigarettes per day for 50 years). We discuss the relationship between transitional cell carcinoma and phenacetin abuse as well as the influence of tobacco-smoking, and review the literature. PMID- 10363147 TI - [A case of valve-like structure in ureter associated with pyonephrosis due to Salmonella infection]. AB - A 7-year-old boy had a fever of 39 degrees C. Abdominal computed tomography (CT) revealed marked left hydronephrosis with hydroureter. Percutaneous nephrostomy was performed. Salmonella infanitis was detected from the drainage urine. Cystourethrography after nephrostomy showed bilateral vesicoureteral reflex (VUR). On the left side, ureteropelvic junction (UPJ) stenosis was found. Left fistelography showed hydronephrosis, but the ureter was not visualized. A mechanism like a valve at the left UPJ was suggested; the bladder urine was able to ascend to the pelvis but not to be drained from the pelvis. The left renal function was not expected to recover from the findings of renal scintigram and CT. Left nephroureterectomy and right anti-VUR operation were performed. The extirpated renal and ureteral specimens revealed a nonpapillary tumorous structure like a valve at the left UPJ. The histological examination of the valve like structure revealed the presence of two muscle layers without an adventitia folded at the UPJ. PMID- 10363148 TI - [A case of true carcinosarcoma in bladder diverticulum]. AB - We report a case of carcinosarcoma arising from a bladder diverticulum. A 71-year old male was referred to our hospital for macroscopic hematuria. Two diverticula were identified in the left wall of the urinary bladder, one of which showed a broad-based tumor. The bladder tumor was resected using a transuretheral approach and the tumor was histologically diagnosed as leiomyosarcoma. The patient underwent partial resection of the bladder including the two diverticula and the tumor. Pathological examination revealed that the resected specimen was composed of three elements, transitional cell carcinoma (G3), squamous cell carcinoma, and leiomyosarcoma. Thus, the patient was diagnosed with carcinosarcoma. He died 5 months after surgery to remove the panperitonitis carcinomatosa. This case is the 38th reported case of bladder carcinosarcoma in Japan. PMID- 10363149 TI - [Bilateral breast metastases from prostatic carcinoma: a case report]. AB - We report a case of bilateral breast metastases from prostatic carcinoma. A 49 year-old man with stage D2 prostate cancer, who had been treated by chemoendocrine therapy and radiotherapy for 2 years, complained of bilateral enlarged breasts. Oral administration of diethylstilbestrol diphosphate was started 2 months before the onset of this symptom. A firm mass that was not tender was palpable beneath the skin without fixation on each side. A needle biopsy of the masses showed poorly differentiated adenocarcinoma with positive immunohistopathological staining for prostate-specific antigen. The masses were diagnosed as metastatic adenocarcinoma of prostate gland origin. The patient died 3 months after the diagnosis of breast metastases. Autopsy revealed diffuse lymphogenous metastatic disease. Metastatic prostatic carcinoma to the breast is uncommon. Breast metastases in this patient might be associated with diffuse lymphogenous metastases as well as increased local blood and lymphatic supply caused by extrinsic estrogens. PMID- 10363150 TI - [Metastatic tumor of renal pelvis and ureter from prostatic cancer: a case report]. AB - An 86-year-old man was admitted to our hospital with a complaint of gross hematuria. Urological examination revealed right hydronephrosis and poorly differentiated adenocarcinoma of the prostate with bone metastases. Abdominal computed tomographic scan and retrograde pyelography showed thickening of the ureteral wall with irregular stenosis in the right upper ureter. Right nephroureterectomy demonstrated diffuse lymphatic infiltration of PSA-positive cancer cells in the submucosa and muscle layer of the ureter as well as renal pelvis. This is the 6th reported case of metastatic ureteral tumor from prostate cancer in the Japanese literature. PMID- 10363151 TI - [A case of pure leiomyoma of the prostate]. AB - Pure leiomyoma of the prostate is very rare. A 67-year-old man was hospitalized with gross hematuria and urinary retention. Drip infusion pyelography revealed a defect of the urinary bladder. By radiological examinations, submucosal tumor of the bladder neck was diagnosed. Transrectal biopsy suggested a benign prostatic tumor histologically. Open resection of the tumor was performed. Histological findings were pure leiomyoma of the prostate. The postoperative course was good and urinary retention was improved remarkably. PMID- 10363152 TI - [A case of genital Paget's disease with severe dermal invasion and early dissemination]. AB - A 65-year-old man was referred to our hospital with a complaint of a scrotal mass which he first noticed 8 months ago. The mass was resected saving genital organs. Pathological diagnosis was extramammary Paget's disease with severe dermal invasion and many nuclear mitoses. The tumor was 15 mm in diameter, and 18 mm in thickness. The bilateral inguinal lymph nodes were dissected and multiple metastases were revealed in the right specimen. Since paraaortic lymph node metastases were detected later 5 cycles of chemotherapy consisting of cyclophosphamide, adriamycin and cisplatin (CAP) followed by radiation therapy were performed. However, only a partial response could be obtained, and multiple brain metastases were revealed on computed tomographic scan. He died 22 months after discovery of this disease. Since extramammary Paget's disease tends to grow slowly and horizontally, cases with such severe dermal invasion and early dissemination as in our case are unusual. PMID- 10363153 TI - [Intrascrotal epidermoid cyst: report of two cases]. AB - A 41-year-old man and a 47-year-old man presented with an intrascrotal mass. The routine studies and ultrasonography showed that the mass was not associated with the testis, epididymis or spermatic cord, but the magnetic resonance imaging was the most useful for making a preoperative diagnosis. Both patients received transscrotal resection of the mass, the histopathological diagnosis of which was epidermoid cyst. A total of 24 cases of epidermoid cyst have been reported in the Japanese literature. PMID- 10363154 TI - [Syphilitic orchitis: a case report]. AB - Syphilitic orchitis is recently a rare disease in Japan. A 75-year-old man was referred to our hospital with the complaint of persistent swelling of the left scrotal contents in spite of prior antibiotic therapy. We suspected a testicular tumor because of lack of pain, and performed high orchiectomy. The specimen showed wide-ranged necrosis with a non-specific inflammatory change of the testis on hematoxylin-eosin stain. After performing analysis using a polymerase chain reaction method, we reached the final diagnosis of syphilitic orchitis. PMID- 10363155 TI - [Statistics on the operations at the Department of Urology, Toyota Memorial Hospital during an 11-year period (1987-1997)]. AB - A clinical statistic survey was carried out on the operations performed at our Hospital from 1987 through 1997. The total number of operations was 3,383 and the number of operations excluding extracorporeal shock wave lithotripsy (ESWL) was 1,672, consisting of 227 (13.6%) operations of the kidney and ureter, 194 (11.6%) operations of the bladder, 481 (28.8%) operations of the prostate and urethra, and 705 (42.2%) operations of the penis and scrotal contents. Since new endourological technology and ESWL were developed for clinical application, the mode of operation has dramatically changed during the last 11 years, trending toward minimally invasive surgery. PMID- 10363156 TI - Application of real-time confocal microscopy for observation of living cells in tissue specimens. AB - Measurement of intracellular Ca2+ concentration ([Ca2+]i) has been a fundamental technique in cell biology. However, most investigations have used cultured or isolated cells as an experimental model, and consequently can provide only limited insight into the mechanisms that operate in tissue in situ. Useful information may be obtained by studying intact tissue specimens. High-speed confocal microscopes that can acquire digital images at video rate have recently been developed. These confocal microscopes which can acquire data in real-time enable [Ca2+]i dynamics of individual cells in intact tissue specimens to be observed. The present paper examines the use of fluorescent microscopy and confocal microscopy for [Ca2+]i imaging of living tissue. We analyzed the dynamics of the duodenal gland, lacrimal gland, intestinal smooth muscles, arterioles, myenteric plexus, and dorsal root ganglion. In these specimens, individual cells exhibited different [Ca2+]i dynamics, and the responses to transmitters/modulators were heterogeneous. In conclusion, real-time imaging provides a useful tool for observing dynamic changes in cells in situ, and it may lead to improve understanding tissue physiology. PMID- 10363157 TI - [Control of tumor-related angiogenesis]. AB - Tumor-related angiogenes is expected to become an important target for improving treatment of cancer, because it plays key roles in tumor growth, invasion and metastasis. We considered that the successful development of such angiostatic treatment depended entirely upon the development of useful anti-angiogenic agents, and attempted to find novel angiogenesis inhibitors by using three in vivo assays, based on an idea of ours. As a result we have demonstrated that different types of agents with low molecular weight including microbial metabolites, cell differentiation modulators like retinoids and steroids, exhibit anti-angiogenic activity, anti-metastatic activity and/or antitumor activity. Taken these findings, an ideal anti-angiogenic agent is discussed. PMID- 10363158 TI - Transcription factor ETS-1 as a molecular target for angiogenesis inhibition. AB - Angiogenesis is a complex phenomenon which includes at least four distinct properties of endothelial cells ECs; degradation of vascular basement membrane and interstitial matrices by proteases, migration, proliferation, and tube formation. We are studying the transcriptional regulation of angiogenesis. We observed that angiogenic growth factors including VEGF and bFGF induced transcription factor ETS-1 in ECs, and ETS-1 converts ECs to angiogenic phenotype by inducing u-PA, MMP-1, MMP-3, MMP-9, and integrin beta 2 as target genes. The elimination of the expression of ETS-1 in ECs inhibited angiogenesis. These results indicate that ETS-1 can be a molecular target for the regulation of angiogenesis. PMID- 10363159 TI - [Regulation of angiogenesis-expression of VEGF receptors]. AB - Angiogenesis, the formation of new blood vessels from pre-existing endothelium, is a crucial process for tumor growth, metastasis and inflammation. Therefore, it is focused on the anti-tumor therapy to prohibit angiogenesis in animal model and clinical studies. Eicosapentaenoic acid (EPA 20: 5,n-3) can restrain tumor growth and inflammation. In this paper, we examined the effects of EPA on tube formation. In EPA-pretreated endothelial cells angiogenesis was attenuated and also proliferation induced by VEGF, but not by b-FGF, was suppressed. The reason why EPA suppressed endothelial cell proliferation induced by VEGF was that EPA selectively inhibited the expression of KDR. As we mentioned, the regulation of angiogenesis in vivo may be involved in the expression of VEGF receptors. PMID- 10363160 TI - Myeloperoxidase positive acute lymphoblastic leukemia cell lines, NALM-30, NALM 31 and NALM-32, carrying Philadelphia chromosome with biphenotypic characteristics. AB - We established three sister cell lines, NALM-30, NALM-31 and NALM-32, with biphenotypic features carrying myeloperoxidase mRNA and protein with complex Philadelphia (Ph) chromosome, t(9;22;10)(q34;q11;q22), from a patient with Ph positive acute leukemia in relapse. Epstein-Barr virus nuclear antigen was negative. The morphological appearance of the cell lines is that of immature lymphoid cells. Expression of myeloid- and lymphoid-associated surface membrane antigens on these cells was detected allowing for the classification of "biphenotypic" leukemia. Immunophenotypically, the established cell lines reported here fulfill the European Group for the Immunological Characterization of Leukemias (EGIL) criteria for B-lineage derivation, however, surface and cytoplasmic immunoglobulin chains were negative. Whereas TGF-beta R (CD105), MCSFR (CD115), SCFR (CD117), IL-4R/IL-13R (CD124) and IL-6R (CD126) were not expressed, the cell lines were mostly positive for IFN-gamma R (CD119), IL-7R (CD127) and FLT-3R (CD135). The NALM-30, NALM-31 and NALM-32 cell lines together with their serial sister cell lines NALM-27 and NALM-28 which were established from the same patient at diagnosis provide unprecedented opportunities for studying a multitude of biological aspects related to normal and neoplastic immature B-lymphocytes. PMID- 10363161 TI - [Basic calponin expressing human neuroblastoma cell line of KP-N-YS]. AB - Recent studies have demonstrated that human neuroblastoma (NB) cell lines have at least two morphological appearance of neuroblastic (N-type) and substrate adhesive (S-type) cells. Our previous study revealed that S-type cells expressed alpha-smooth muscle actin and/or desmin, suggesting the smooth muscle cell characteristics of S-type cells. In the present study, a new human NB cell line, KP-N-YS, was established from bone marrow metastasis of a four-year-old boy with advanced NB, originating from the left adrenal gland. Subsequently N-type cell clone (YS1n) and S-type cell clone (YS2s) was isolated from the parent cell line. Parent and clones cell lines were identified as NB by surface membrane antigen and cytoskeletal protein analysis, and these cell lines were demonstrated as common progenitors by chromosomal analysis. Furthermore the presence of basic calponin were determined by indirect immunofluorescence, Western blot, as well as by RT-PCR (reverse transcriptase-polymerase chain reaction). Our demonstration of basic calponin not in N-type cells, but in S-type cells supports the plausible smooth muscle cell characteristics of this NB cell line. PMID- 10363162 TI - Expression of cathepsin B-like enzyme activity in cell lysate from cultured human normal keratinocytes. AB - In the present study, we have found that the cell lysate from cultured human normal keratinocytes from foreskin (HFKs) hydrolyzed alpha-N-benzoyl-DL-arginine beta-naphthylamide (BANA), and the BANA hydrolysis occurred most under conditions of 37 degrees C and pH 6.0. This activity was strongly inhibited by leupeptin, which is an inhibitor to cathepsin B. These results suggested that the cell lysate from cultured HFKs contained cathepsin B-like enzyme activity. This is the first report to demonstrate that cathepsin B-like enzyme activity was expressed in the cell lysate from human normal keratinocytes. PMID- 10363163 TI - The human poly(ADP-ribose) glycohydrolase maps to chromosome 10q11.23-21.1 by fluorescence in situ hybridization. AB - Poly(ADP-ribose) glycohydrolase (PARG) digests poly(ADP-ribose), which is synthesized by poly(ADP-ribose) polymerase (PARP) after DNA damage. We mapped the human poly(ADP-ribose) glycohydrolase gene to chromosome 10q11.23-21.1 by fluorescence in situ hybridization analysis. Since chromosomal rearrangements in thyroid papillary carcinoma and loss of heterozygosity in glioblastoma are frequently observed in this region, genetic alteration of PARG could be implicated in these diseases. PMID- 10363165 TI - Variability of intracellular concentrations of basic fibroblast growth factor in cultured human gliomas. AB - BACKGROUND: Basic fibroblast growth factor (bFGF) is a small peptide with angiogenic and mitogenic properties that supports the growth and proliteration of human malignant glial tumors in vitro and in vivo in an autocrine fashion. The purpose of this study was to examine the potential relationship between intracellular bFGF concentrations and grade of glial malignancy using a monolayer cell culture system. MATERIALS AND METHODS: Samples from 13 histopathologically verified astrocytic brain tumors and two non-tumorous astrocyte specimens were grown in tissue culture and examined both early and late after explantation together with a bFGF-producing reference cell line. RESULTS: Elevated intracellular concentrations of bFGF were noted in the reference line as well as two of five other glioblastoma multiforme-derived cell cultules, three of five anaplastic glioma-derived cell cultures, and two of three astrocytoma-derived cell cultures. The cells derived from non-tumorous astrocyte specimens expressed low concentrations of bFGF. CONCLUSION: This study demonstrates that overlap exists between the grade ot glial malignancy and intracellular bFGF levels. PMID- 10363166 TI - The effect of dexamethasone on tissue fibrinolytic system in male and female rats. AB - BACKGROUND: Dexamethasone in low and high doses affects blood fibrinolytic activity both in animals and humans. In this study the effect of a high dose of dexamethasone on plasminogen activator activity (PAA), t-PA antigen level, plasminogen activator inhibition (PAI) and plasmin inhibition (Pl) in rat heart, brain, liver, lungs and kidneys was investigated in both sexes. MATERIALS AND METHODS: Twenty male and twenty female adult Wistar rats were used. Dexamethasone was administered as a single intraperitoneal injection (3mg/kg/day) in rats, once daily, for a period of 5 consecutive days. t-PA antigen level was assayed by an enzyme-linked immunoabsorbant assay method. PAA, PAI and Pl were determined by spectrophotometric methods. The plasminogen used was isolated from rat plasma. RESULTS: Dexamethasone induced variable changes in the fibrinolytic parameters in rat heart, brain and liver of both sexes; in lungs and kidneys dexamethasone had no effect. CONCLUSION: These changes of PAA, PAI and t-PA antigen level in heart, brain and liver induced by dexamethasone might be of importance regarding the involvement of glucocorticoids and plasminogen activators/plasmin in many pathophysiological conditions. PMID- 10363167 TI - Cancer induction studies using different administrations of benzenediazonium sulfate in mice. AB - Benzenediazonium sulfate (BD) was administered as 10 weekly subcutaneous injections at 25 micrograms/g b.w. and as 52 weekly oral gavages at 100 micrograms/g b.w. to Swiss mice, starting at 6 weeks of age. The subcutaneous administration induced tumors in the subcutis with an incidence of 8% in females. The oral treatment gave rise to lung tumors with incidences of 52% in females and 62% in males. In the untreated control female mice, no subcutaneous tissue tumor was observed, but the incidences of lung tumors were 28% in females and 38% in males. Histopathologically, the neoplasms were classified as fibrosarcomas of the subcutis and adenomas and adenocarcinomas of the lungs. In an earlier experiment, BD induced high incidences of subcutaneous tissue tumors in the same species when it was administered as 26 weekly subcutaneous injections at 10 micrograms/g. This indicates the length of treatment is paramount to the dose of carcinogen. The oral route, even though it was carcinogenic in the lungs, failed to elicit the development of cancer in the glandular stomach. PMID- 10363168 TI - Sonographic estimation of liver tumor development induced by oral administration of thioacetamide in rat. AB - Animal models for various types of cancer are of great help in the study of tumors and antitumor effects. Subcutaneous models have been widely utilized because they can be produced easily by subcutaneously implanting tumor cells into animals. Although subcutaneous models are very convenient for examining tumor development, they are definitely different from clinical manifestations of original tumors. In orthotopic animal models for internal tumors, however, it is difficult to examine tumor development without sacrificing animals. We demonstrate here that the occurrence and growth of liver tumors induced by oral administration of thioacetamide into rats were clearly observable by ultrasonography, and that the sonographic estimation was accurate. It was observed sonographically that the number and volume of liver tumors increased proportionately with TAA treatment periods. These results indicate that sonography is a useful and non-invasive method to investigate liver tumor development in rats. PMID- 10363169 TI - Differences of milk-transmitted murine mammary tumor virus (MMTV) among mouse strains. AB - To clarify the oncological differences of milk-transmitted Murine Mammary Tumor Virus (MMTV) in various strains, BALB/c mice were foster-nursed on C3H/He, GR, DDD, DDD/1-Mtv-2/Mtv-2 and FM strains and observed for the development and morphology of mammary tumors, followed by testing tumors for pregnancy dependence (PD) after transplantation. MMTV were different in tumorigenicity and morphology and PD of induced tumors among the mouse strains. These differences of MMTV appeared not to be parallel with the difference in specificity of superantigens which are encoded by MMTV on the previous report. Interestingly, host factors influenced the properties of MMTV produced by the same endogenous Mtv gene. PMID- 10363170 TI - Acute and late effects of tritium beta particles on rats exposed to tritiated water as infants. AB - Lethality and physiological disorders induced by tritium beta particles were studied in rats. Newborn CD/Crj rats received i.p. HTO saline at 0, 8.14, 16.28, 24.42 and 32.56 MBq/g body weight (BW). At four weeks of age, HTO over 24.42 MBq/g BW was fatal, especially in male rats, but it was not obvious under 16.28 MBq/g BW. Surviving female rats were weaned at four weeks of age and rats were monitored for heart rate and blood pressure (systolic, mean and diastolic) at 13, 23 and 30 weeks of age and sacrificed. The blood pressure significantly increased with the administered doses of HTO. Sclerotic kidneys were frequently observed in hypertensive rats exposed to HTO over 24.42 MBq/g BW. Thickened intermediate layer of artery, infiltrating lymphocytes and atrophic mesangiums were observed in sclerotic kidneys. These results indicate that internal exposure to HTO as infants was induced consequent hypertension in association with sclerotic kidney at sublethal doses. PMID- 10363171 TI - Surgical stress induces a marked and sustained increase of adrenal androgen secretion in postmenopausal women. AB - We analyzed the secretion of adrenal androgens in response to surgical stress in eight (8) postmenopausal women. ACTH, cortisol (F) and adrenal androgens, such as dehydroepiandrosterone (DHEA), delta 4-androstendione (delta 4-A), dehydroepiandrosterone sulfate (DHEA-S) and testosterone (T) were measured at 08:00 and 20:00 hr the day before and for three consecutive days after operation, as well as at 0, 15, 30, 60, 120 minutes during cholocystectomy. Basal levels of ACTH, F, DHEA, delta 4-A, DHEA-S and T were found within the normal range for this age group before surgery. During surgery the ACTH was significantly increased, reaching a peak value at 30 min after surgery initiation. F, DHEA and delta 4-A were significantly increased during and after surgery, returning to pre surgery levels by the third day post-surgery. DHEA-S levels did not increase during surgery but was found significantly increased the day after surgery, returning to presurgical levels two days later. We conclude that surgical stress can induce adrenal androgen hypersecretion during and within the early days post surgery. Because the adrenal androgens levels are declining and respond suboptimally to exogenous corticotropin releasing hormone (CRH) or ACTH bolus injection in aging women, it is conceivable that this remarkable response of adrenal androgens to surgical stress is probably of biological significance and conceivably mediated by a CRH/ACTH-independent mechanism. PMID- 10363172 TI - Estrogen receptor content in adenovirus type 9-induced rat mammary tumors. AB - Hormonal factors play an important role in the induction of mammary tumors and tumor-like lesions in adenovirus type 9-inoculated W/Fu rats. Primary Ad 9 induced fibroadenomas contained significantly higher amounts of estrogen receptor (determined by means of enzyme immunoassay) in comparison to normal breast tissue (p = 0.01**) and "spontaneous" fibroadenomas (p = 0.03*), used as control tissues. The receptor content of serially isografted virus-induced fibroadenomas did not differ significantly from the two types of control tissue. The findings suggest that changes in the estrogen receptor level are of importance in the tumor induction process, but also that additional factors are required for the preservation of tumor characteristics as well as for lipoma induction. PMID- 10363173 TI - Prevention of oral diseases by polyphenols (review). AB - This review summarizes the current data on the effects of natural products on various oral diseases, together with their basic biological activities. We have focused on polyphenols and their secondary metabolites, such as tannins, lignins and flavonoids, and their modulating factors, including saliva proline-rich proteins. These substances are found in a wide variety of plant sources such as vegetables, herbs, nuts and teas, and effectively reduce the incidence of carcinogenesis in the oral cavity, inhibit plaque growth and adsorption of oral bacteria, and inhibit the replication of various viruses. The mechanism of their action includes: the direct inactivation of the bacteria and viruses, inhibition of their replication enzymes, induction of apoptosis in tumor cells, stimulation of monocytes/macrophages to produce cytokines, and stimulation of myeloperoxidase dependent iodination of neutrophiles. Polyphenols showed biphasic actions, acting as antioxidants at lower doses, but acting as prooxidants at higher doses. The development and progression of oral diseases might be modified not only by these natural products, but also by interaction with saliva, proline-rich proteins, antioxidants, metals and dental materials. PMID- 10363174 TI - Detection and clinical significance of plasma glutathione-S-transferases in dogs with lymphoma. AB - BACKGROUND: The objective of this study was to determine if glutathione-S transferases were detectable in the plasma of dogs and to determine if concentrations of the a- and pi-subtypes were related with tumor response to single agent anthracycline (e.g., doxorubicin) chemotherapy in dogs with lymphoma. MATERIALS AND METHODS: Plasma was obtained from 10 healthy, normal dogs and from 11 dogs with lymphoma before treatment, 3 weeks after 1 dose of doxorubicin and every 3 weeks thereafter until relapse (the physical detection of recurrent and enlarged peripheral lymph nodes). Plasma concentration of alpha and pi-GST was determined by use of an ELISA technique with well plates pre-coated with IgG[anti-Canine alpha-GST or anti-Human pi-GST]. RESULTS: Mean plasma alpha GST concentrations did not significantly decline after 1 dose of doxorubicin chemotherapy; however, mean plasma alpha-GST concentrations were markedly increased (p < 0.05) at the time of relapse (the physical detection of recurrent and enlarged peripheral lymph nodes). CONCLUSIONS: In this study we show that a relationship exists between the plasma alpha-GST concentration and the clinical response of dogs with lymphoma to doxorubicin chemotherapy. PMID- 10363175 TI - Increased level of serum hepatocyte growth factor/scatter factor in liver cancer is associated with tumor metastasis. AB - Hepatocyte growth factor/scatter factor (HGF/SF) has been shown to play an important role in tumor migration and metastasis. We therefore investigated the relationship between HGF/SF expression and metastasis of human liver tumors. The serum HGF/SF levels in 41 patients with primary hepatocellular carcinoma (HCC; 22 patients with metastasis, 19 patients without metastasis); 4 patients with benign hepatic tumor, 4 patients with secondary hepatic carcinoma, and 12 healthy blood donors, were measured by sandwich enzyme-linked immunosorbent assay (ELISA) in this study. Our results show that liver tumor patients have significantly higher serum levels of HGF/SF compared to healthy blood donors. However, there is no difference between the serum HGF/SF levels in patients with primary HCC and patients with secondary HCC or benign hepatic tumors. In addition, we measured significantly higher levels of HGF/SF in serum from HCC patients with metastasis compared to HCC patients without metastasis, indicating that the elevations in serum HGF/SF level correlated positively with the tumor metastasis in human HCC. These findings appear to suggest that HGF/SF may be a useful serological biomarker for clinical diagnosis and follow-up of HCC metastasis, and suggest that larger scale studies in patients with hepatomas are warranted. PMID- 10363176 TI - A safe, effective in vivo gene therapy for melanoma using tyrosinase promoter driven cytosine deaminase gene. AB - This study was designed to develop a safe, effective gene therapy for disseminated melanoma. We constructed retroviral vectors containing a tyrosinase promoter-cytosine deaminase expression cassette (Tyr/CD), and demonstrated that the tyrosinase promoter conferred a selective expression of cytosine deaminase (CD) gene in B16 melanoma cells, especially when the Tyr/CD cassette inserted in 3'LTR region of a retroviral vector. In vivo gene therapy for the intraperitoneally disseminated melanoma using Tyr/CD retrovirus-producing cells and 5-fluorocytosine (5-FC) showed that retroviruses produced in situ were capable of infecting tumor xenografts and bone marrow cells in animal model, and survival rates were prolonged significantly as compared with those treated with CD2 retrovirus-producing cells and 5-FC. Importantly, the treatment-related bone marrow suppression was not observed in the former treatment, while profound bone marrow suppression was observed in the latter treatment. In vivo gene therapy using retrovirus-producing cells containing suicide gene under the control of a tissue-specific promoter and 5-FC administration is safer and more effective for the treatment of disseminated melanoma, as compared with retrovirus-producing cells containing the gene under the control of a universal promoter and 5-FC. PMID- 10363177 TI - Detection of SIV DNA in rhesus macaque polymorphonuclear neutrophils. AB - The extent of infection of monkey polymorphonuclear neutrophils (PMN) by simian immunodeficiency virus (SIV) has not yet been determined. Using the polymerase chain reaction (PCR) technique, we detected the presence of SIVmac239 DNA in rhesus macaque-derived PMN after 24 hrs of in vitro incubation of the cells with SIVmac239. Infection by SIVmac239 also down-regulated the expression of the bcl-2 apoptosis-blocking gene in the infected PMN. These SIVmac239-induced PMN intracellular alterations were correlated with an accelerated decrease in PMN viability over a period of 120 hrs compared to non-infected PMN. Evidence of chromatin condensation characteristic of programmed cell death (apoptosis) was also observed in SIVmac239-infected PMN. The results of this study provide a mechanism for the reduced chemotaxis/phagocytosis activities of PMN of SIVmac239 infected macaques and suggest that PMN is one of the target cells for SIVmac239 infection. PMID- 10363178 TI - Tissue levels of chemotherapeutic agents for hepatic metastasis during hepatic arterial and portal injection. AB - Hepatic metastasis is one of the most important prognostic factors in digestive organ cancer, and hepatic arterial infusion is aggressively performed for therapy of nonresectable metastatic liver cancer. Although comparatively high response rates have been attained in some cases, this treatment has been ineffective in not a few cases because these metastatic tumors are frequently hypovascular in nature. To develop better methods of administering chemotherapeutic agents, we performed basic experiments concerning intraportal administration which has been regarded as having a generally negative effect, focusing on a report indicating that portal supply is dominant along the borders of metastatic liver cancer tumors. VX2 carcinoma cells were inoculated into the hepatic parenchyma beneath the capsule of juvenile Japanese white rabbits. Drugs were infused 2 weeks after the inoculation, then tissue and blood were sequentially sampled. Mitomycin C (1.7 mg/kg) was infused either by bolus injection to the hepatic artery (arterial infusion group) or by bolus injection to the portal vein (portal infusion group). Five-fluorouracil (9.5 mg/kg) and Cisplatin (1.6 mg/kg) were likewise infused continuously over 60 min, and tissue levels of the drugs were compared between the two groups. Mitomycin C and 5-fluorouracil levels were measured by HPLC and Cisplatin levels were measured by atomic absorption spectrophotometry. As a result, the levels of every drug in VX2 tumor tissue did not significantly differ between the arterial infusion group and the portal infusion group, while the levels were significantly higher than those in the intravenous infusion group. Using portal infusion, we observed a drug transition which was not inferior to that of arterial infusion, suggesting that an imported antitumoral effect may be obtained with this method compared with intravenous infusion. PMID- 10363179 TI - Memory and therapeutic action. PMID- 10363180 TI - 'Dreams that turn over a page'. Integration dreams with paradoxical regressive content. AB - The subject of this paper is 'dreams that turn over a page', whose primitive anxiety-inducing content frightens the dreamer, although the psychoanalyst sees them as a sign of progress in psychic integration despite their regressive appearance. This thesis is illustrated by clinical examples. Because such anxiety dreams typically occur at a time of integration, the author considers that the analyst must interpret them as showing that the patient is now able to accept hitherto unrepresentable parts of himself, while the dream content proper should be interpreted at a second stage once the anxiety has subsided. The author postulates that these seemingly regressive dreams are a token of progress because they occur at privileged points in the transference when projective identification is giving way to introjection. The reintegration of previously expelled fragments causes anxiety but also gives the dreamer a sense of inner cohesion, while at the same time accounting for the particular clarity and coherence of these dreams. The author compares his concept of dreams that turn over a page with similar notions in the literature and contends that such dreams retrospectively illuminate changes in the dreamer's intrapsychic conflicts on a more elementary level of unconscious fantasy than the classical approach would suggest. PMID- 10363181 TI - Speaking in the claustrum: the psychodynamics of stuttering. AB - The author outlines the psychoanalytic theory of stuttering and, discussing material from the analysis of a stutterer and its transference and countertransference processes, puts forward a new hypothesis of the psychodynamics of stuttering in conjunction with Meltzer's theory of the claustrum. He argues that the stutterer is working out intolerable experiences of separation from the primary object and a resulting catastrophic experience of the oedipal situation through an unconscious fantasy in which anal qualities are conferred on the internal maternal object by a predominating hatred. The intrusive identification of parts of the self in the maternal rectum gives rise to a claustrophobic experiential world in which all obstacles that are encountered between self and object must be eliminated. The anal-sadistic object space of the claustrum is projected on to the external object space and thus also on to the mouth as the origin of the sound envelope, where it produces both a lifeless sound envelope and a torn content, i.e. stuttered sounds, words and sentences. Correspondingly, a dead speech melody and broken words have their parallels in object relations that are characterised by an attack on linking and by psychic withdrawal. PMID- 10363182 TI - A language of dreaming: dreams of an Amazonian insomniac. AB - The author describes a series of psychoanalytic interviews conducted on the banks of an Amazonian stream, in the course of which a lively old Parintintin woman, wife of a chief, acknowledges and comes to terms with her lifelong insomnia. When she sleeps, she is plagued with anxious dreams; and her dreams take her back to her childhood as the daughter of a prominent chief and to memories of the strife between her parents. The way she talks about her dreams illustrates how cultural beliefs may facilitate the communication of dreams and introspective reflection on them, but may at the same time intensify anxieties. This article explores the cultural context of psychoanalytic understanding of dreams: how are dreams shaped by the dreamer's cultural beliefs about them and ways of interpreting them? How are they affected by the social uses the dreamer makes of them? The work also examines relationships formed in anthropological field-work, especially the separation that occurs when the anthropologist leaves (not unlike termination of an analysis)--an issue inadequately discussed in the anthropological literature and in the psychoanalytic literature on cross-cultural research. PMID- 10363183 TI - On discriminating and not discriminating between affect and representation. AB - The topic involves two issues. They can be related either to different clinical pictures or to divergent opinions about the same clinical facts. But the above mentioned opposition can be found in Freud's work. Listening in analysis differentiates between situations where the distinction of affect and representation is blurred within the general frame of communication and others where this distinction imposes itself because of the critical or chronic character of the predominance of affect. Problems about the discrimination between affect and representation in the unconscious are elucidated in the light of their structural differences. The notion of the psychical representative of the drive announces the reference to the further notion of instinctual impulse. Later on, in Freud's theory, the notion of instinctual impulse has encompassed the distinction between affects and representation. Contemporary authors have, in their vast majority, chosen to refer preferably to object relationships. Nevertheless, the problems raised by Freud remain unanswered. It is with the clinical picture of borderline personality disorders that the lack of discrimination between affect and representation becomes evident. A detailed description is presented of the forms of transference where the irrepresentable prevails, accompanied by feelings of being overwhelmed, repetition compulsion, acting out and somatic reactions. In the countertransference, the affects of helplessness, despair and even the impossibility of grasping the meaning of the patient's communication are frequently met in the analyst. In these clinical pictures, intermediary formations, i.e. psychic organisations where primary processes structures the unconscious, are impaired. One is struck by the limitation of the capacities of representation and by the importance of phenomena related to negative hallucinations specially focused on thought processes, The communication is frequently self-contradictory, the transference oscillating between disembodiment and primitive fusion. The object has a paradoxical status; it is at the same time everything and nothing, the transference being incomprehensible and unanalysable. Different hypotheses are presented to explain the importance of destruction. Finally, the author presents a hypothesis for the deficiency of these intermediary formations because of the absence of continuing cathexis from the primary object at certain important moments of the relationship. It is the establishment of theses intermediary formations that will permit the acceptance of the separation from the object, the child now being able to rely on his own internal creations. However important the influences of the relationship to the other in the genesis of affects, the internal orientation towards the body of the psychic processes is what constitutes the core of the primordial psychic world. PMID- 10363184 TI - Moving ahead: integrating influences of affective processes for development and for psychoanalysis. AB - The author argues that psychoanalytic clinical thinking has evolved towards an organisational model of affect and that multidisciplinary research broadens this thinking. Integrative influences of affective processes have been given little attention in psychoanalytic theory. Such influences are reviewed using examples from research in early development. Affective processes are shown to provide integrative influences across systems in an individual's development, facilitating developmental change, as well as developmental continuity. In a related vein, affective processes provide incentives for the development of both individuality and connectedness with others. The notion of an 'affective core of self' is updated and the important psychoanalytic idea of emotion schemas of self in relation to others is reviewed. Integrating influences of affective processes in psychoanalysis operate mainly non-consciously and are in need of further study. PMID- 10363185 TI - Affect, language and communication: loose ends. AB - The author begins by enquiring whether the three elements of his subject matter are intended to be discussed separately or in terms of the links between them. He goes on to consider the role of language in the development of psychoanalysis from Freud on. Notwithstanding the fruitful mutual influences of linguistics and psychoanalysis, he draws attention to the dangers of a one-sidedly linguistic approach to psychoanalytic theory and clinical practice. Turning to affect, the author reviews Freud's early neurophysiological conceptions and the possible distinctions between affect, emotion and feeling as reflected in the contributions of later workers. The aspects of anxiety, trauma, object relations, transference and infant development are touched upon and quantitative and metapsychological considerations are adduced. The author's starting point for his treatment of communication is Freud's well-known telephone simile. He goes on to discuss subsequent work on mother-baby communication in early childhood, communication as semiology, and communication through acting out and enactment; this section ends with a note on the etymology of the word communication. The author concludes by drawing attention to the importance in analysis of experience, in which the three entities of language, affect and communication are combined. Having failed to answer the question with which he began, he is left with the loose ends of his title. PMID- 10363186 TI - Cognitive linguistics. PMID- 10363187 TI - Shall we ever know the whole truth about projective identification? PMID- 10363189 TI - 'The recovered memories controversy'. PMID- 10363188 TI - Who influenced whom? And how?--a brief series of notes on E. Weiss, M. Klein (and I. Svevo) and the so-called 'origins' of 'projective and introjective identification'. PMID- 10363192 TI - 'Psychoanalysis and cognitive-evolutionary psychology: an attempt at integration' by Paolo Migone and Giovanni Liotti. PMID- 10363191 TI - Renik replies to Cavell. PMID- 10363193 TI - Inflammatory bowel disease and osteoporosis. PMID- 10363190 TI - 'The ambulatorium: Freud's free clinic in Vienna'. PMID- 10363194 TI - Helicobacter pylori increases the risk of peptic ulcer bleeding: a case-control study. AB - AIM: Aim of the present case-control study was to establish whether Helicobacter pylori increases the risk of ulcer bleeding. PATIENTS AND METHODS: All patients presenting with upper gastrointestinal bleeding between November 1994 and November 1995 were prospectively investigated and compared with hospital controls matched for age, sex, and race. We evaluated the frequency of Helicobacter pylori infection, intake of aspirin or non-steroidal anti-inflammatory drugs, use of alcohol, and smoking habits in patients and controls. RESULTS: Included in the study were 128 patients. In 72 patients, the source of bleeding was a peptic ulcer (duodenal ulcer: n = 33; gastric ulcer: n = 39). Ulcer patients were more frequently infected by Helicobacter pylori than controls (72% vs 42%; p < 0.001) while the incidence of infection was similar in patients with non-ulcer bleeding and controls (52% vs 46%; p = 0.59). Conditional multiple logistic regression analysis showed that Helicobacter pylori infection (odds ratio, 3.3 [Confidence interval, 1.5 to 7.0]; p = 0.002) and regular use of alcohol (odds ratio, 3.1 [Confidence interval, 1.0 to 9.0]; p = 0.041) increased the risk of peptic ulcer bleeding while previous intake of aspirin (> 100 mg) or non-steroidal anti inflammatory drugs independently increased the risk of bleeding only in the case of gastric ulcer (odds ratio, 8.1 [Confidence interval 1.2 to 56.6]; p = 0.034). CONCLUSIONS: Helicobacter pylori infection increases the risk of peptic ulcer bleeding. Our results suggest that Helicobacter pylori and non-steroidal anti inflammatory drugs are independent risk factors for peptic ulcer bleeding. PMID- 10363195 TI - Helicobacter pylori and non-steroidal anti-inflammatory drugs: ulcers and bleeding ulcers. PMID- 10363196 TI - Limitations of the faecal occult blood test in screening for colorectal cancer. AB - BACKGROUND: A screening test should be acceptable, safe, simple, accurate, reliable, effective, and inexpensive. Screening for colorectal cancer with the faecal occult blood test is done to reduce the incidence and mortality from colorectal cancer. How does this test measure up to these requirements? METHODS: The characteristics of faecal occult blood test are described by means of clinical epidemiology, analysing its compliance, sensitivity, specificity, positive predictive value, and its test performance in Bernoulli trials. A decision tree and a Markov model are used to compare the cost-effectiveness of screening strategies involving faecal occult blood test and colonoscopy. RESULTS: After 5-10 years, patient compliance in a faecal occult blood test screening programme falls below 50%. Over 80% of the patients are likely to leave the programme before its completion. Although the test itself may seem safe and simple, the high rate of false-positive outcomes exposes many subjects to the potential complications of colonoscopy. The high rate of false-negative tests gives patients with colorectal cancer a false sense of security and delays their proper diagnostic work-up. In populations with low prevalence rates of colorectal cancer, faecal occult blood test becomes very inaccurate in diagnosing colorectal cancer, as its positive predictive value falls below 5%. Its long-term test performance is unreliable in that it comes to depend on the frequency, with which the test is repeated. Any negative or positive test result can be achieved by varying the frequency of test repetition. Screening by colonoscopy every five or ten years is more cost-effective than screening by annual faecal occult blood test in preventing the occurrence of colorectal cancer and its associated mortality. CONCLUSIONS: Screening strategies for colorectal cancer involving faecal occult blood test should be abandoned. PMID- 10363197 TI - Screening for colorectal neoplasia. PMID- 10363198 TI - Pilot study of triple antiviral therapy for chronic hepatitis C in interferon alpha non-responders. AB - BACKGROUND: No effective therapy exists for interferon non-responding chronic hepatitis C patients. AIMS: Pilot study evaluating the potential efficacy and safety of triple antiviral therapy in interferon-alpha non-responders. PATIENTS AND METHODS: Twenty consecutive adult patients with chronic hepatitis C who had failed to respond to a 6-month course of interferon alpha were randomly assigned to receive a combination of interferon alpha + oral ribavirin (double therapy), or the same combination + oral amantadine (triple therapy), for 6 months. RESULTS: By the end of therapy, normal alanine transaminase (biochemical response) was obtained in 2 out of 10 patients on double therapy but in 7 out of 10 on triple therapy (p < 0.05), and negative serum hepatitis C virus (HCV) RNA (virological response) occurred in 1 out of 10 patients on double therapy but in 7 out of 10 patients on triple therapy (p < 0.01). Six months after therapy, biochemical response was sustained in 1 (double therapy) and 4 patients (triple therapy), respectively, and the virological response was sustained in no patient on double therapy but in 3 patients on triple therapy. CONCLUSIONS: Triple antiviral therapy seems to be able to induce biochemical and virological responses in interferon alpha non-responders with chronic hepatitis C. PMID- 10363200 TI - Aging and the gastrointestinal tract. AB - The aim of the present review is to summarize the most recent progress in gastroenterological topics, particularly of the upper gastrointestinal tract, which are of special interest in the elderly. The changes in oesophageal function, particularly disorders of motility, may explain, only in part, the unique clinical characteristics of oesophageal pathologies in the elderly. Dysphagia and gastro-oesophageal reflux disease present diagnostic, clinical and therapeutic characteristics that need to be studied with attention to avoid eventual disability, an impairment of nutritional status and a reduction in the quality of life. Aging, per se, does not significantly modify gastric aggressive factors, however, a selective and specific reduction in some gastric defensive mechanisms seems to occur with aging. The prevention of gastric mucosal injury, particularly that due to drugs, requires a better understanding of these age related changes. Helicobacter pylori infection in the elderly presents peculiar epidemiological aspects particularly for subjects living in nursing homes. An understanding of Helicobacter pylori-related histological modifications of the gastric mucosa, particularly intestinal metaplasia, gastric atrophy and gastric cancer, the incidence of which seems to be both age- and Helicobacter pylori related, is greatly needed. Moreover, some diagnostic and therapeutic aspects of Helicobacter pylori infection, i.e., the role of serology and the efficacy, side effects and compliance of drug therapies, are specific for the elderly and require a unique clinical approach. Bleeding is dramatically more frequent in this population. The identification of risk factors, i.e., drugs, pathophysiological mechanisms, i.e., the possible relationship between non steroidal anti-inflammatory drugs and Helicobacter pylori infection, along with the clinical presentation in the elderly patient, must be the foundation of preventive medical care in geriatric gastroenterology. PMID- 10363199 TI - Combination therapy for chronic hepatitis C. PMID- 10363201 TI - What is the current role of endoscopic ultrasonography in oesophageal cancer? AB - Endoscopic ultrasonography is, probably, the most significant progress in gastrointestinal endoscopy of the last few years, and it is on the way to becoming the gold standard for pre-treatment staging of oesophageal carcinoma. Once computed tomography scan has ruled out distant metastases or evident local invasion, endoscopic ultrasonography plays a key role in an adjunctive evaluation and planning of treatment. The diagnostic accuracy of endoscopic ultrasonography in oesophageal cancer is excellent. Its main limitations are: poor differentiation between early mucosal and submucosal cancer, insufficient accuracy in detecting early lymph node invasion, and lack of complete tumour evaluation in cases of high-grade strictures. The role of endoscopic ultrasonography in post-treatment assessment is still limited by some artifacts. However, all these limitations will soon be overcome by some technical improvements. PMID- 10363202 TI - Apoptosis and gastrointestinal tract. AB - The gastrointestinal tract is characterized by a rapid proliferation of stem cells that differentiate to become terminal mature cells and ultimately die through a genetically programmed form of cell death, termed apoptosis, which is responsible for maintaining of tissue size. Apoptosis has also been shown to play an important role in the pathophysiology of several gastrointestinal diseases. The development of many infectious and immune-mediated diseases, such as gastritis, coeliac disease, inflammatory bowel diseases, may be triggered by the prevalence of pro-apoptotic signals, whereas prolonged cell survival, due to apoptosis inhibition, may give rise to neoplastic clones. Elucidation of the biochemical pathways and of specific proteins regulating apoptosis may provide a remarkable opportunity to manipulate the life and death decisions of the gastrointestinal cells and to develop new therapeutic strategies. This review will deal with the mechanisms potentially involved in apoptosis and with the clinical relevance of this phenomenon in gastrointestinal diseases. PMID- 10363203 TI - Gene regulation in hepatic stellate cell. AB - Hepatic stellate cells are now recognized as the major source of extracellular matrix in hepatic fibrosis. Following liver injury the hepatic stellate cell changes from a quiescent to an activated cell. The activation process includes an increased proliferation rate, a phenotypic change to a myofibroblast-like cell, loss of vitamin A stores, increased extra-cellular matrix protein synthesis and contractility. Furthermore, hepatic stellate cells have been implicated in hepatic inflammation through their ability to secrete cytokines and chemokines. Here, we review the literature on the molecular pathogenesis of hepatic stellate cells activation with emphasis on the most recent findings. The reviewed topics include transcriptional and post-transcriptional regulation of the genes encoding type I collagen in hepatic stellate cells; the role of the transcription factor nuclear factor Kappa B in the hepatic stellate cell activation; focal adhesion kinase and integrin-mediated signal transduction in hepatic stellate cell, and apoptosis in hepatic stellate cells. New insight into hepatic stellate cell activation and death may lead to the development of novel therapies for hepatic fibrosis. PMID- 10363204 TI - Laboratory role in the management of hospital acquired infections. AB - The microbiology laboratory has many important roles. It must collaborate with the infection control team on the investigations of outbreaks. During outbreaks, it must save relevant samples, look for reservoirs and undertake typing techniques, all of which should be timely. New technology should be available to detect, identify and characterize micro-organisms. Molecular biological techniques have enhanced the speed and sensitivity of detection methods and have allowed the laboratory to identify organisms that do not grow or grow slowly in culture. Molecular techniques also enable the microbiologist to identify antibiotic resistance genes and to 'fingerprint' hospital organisms, thereby facilitating studies of nosocomial transmission. PMID- 10363205 TI - Assessing the role of prophylactic antibiotics in clean surgery. PMID- 10363206 TI - Microbiological evaluation of four enteral feeding systems which have been deliberately subjected to faulty handling procedures. AB - The present study was designed to investigate the levels of contamination in four currently used 1000 mL, 'ready-to-hang', enteral feeding systems--Osmolite (Ross Ready-To-Hang), Steriflo, Dripac-flex and Easybag, when faulty handling procedures were used during assembly of the systems. The top of the nutrient container and the proximal (container) end of the pump set of each system were touched during assembly by a researcher whose hands had been deliberately contaminated with Klebsiella aerogenes. Once assembled systems were run continuously for 24 h delivering 1000 mL of feed. Feed samples for microbiological analysis were taken from the distal (patient) end of the feeding tube at 0 h and 24 h and from the feed remaining in the nutrient container at the end of administration (24 h). Five systems of each type were run. Five controls were also run for each type of system, where all procedures were carried out wearing sterile gloves. Eighty-seven percent of feed samples collected from the Osmolite systems and 80% of those from the Steriflo systems were found to contain K. aerogenes, with 13% of feed samples from both systems containing > or = 10(4) cfu/mL, a level of contamination, considered by many, as that above which feed is unacceptable for patient consumption. The percentage of feed systems containing the test organism was much lower in the Dripac-flex and Easybag systems, with K. aerogenes being detected in 27% and 13% of samples respectively. No feed samples from either of these systems contained > or = 10(4) cfu/mL. From the results it can be concluded that deviation from the manufacturers instructions when assembling enteral feeding systems can lead to bacterial contamination of these systems. The results also highlight the effect that system design, such as recessed pump set spikes and recessed nutrient container seals (both of which prevent care workers accidentally touching parts of the feeding system which may come into contact with the feed) have on reducing the number of bacteria gaining entry to the feed in the systems. PMID- 10363207 TI - Endemic nosocomial gram-negative bacteraemias resulting from contamination of intravenous heparin infusions. AB - Following several cases of Gram-negative bacteraemia secondary to intravenous heparin infusion contamination, we retrospectively reviewed nosocomial bacteraemias associated with heparin infusions at our institution. Thirty-one episodes of heparin-infusion related bacteraemia occurred in 30 patients over a 23-month period affecting 2% patients receiving heparin infusions for more than 48 h. Gram-negative bacteria were responsible for all bacteraemias. The care of infusions during clinical use was prospectively surveyed, revealing that approximately 20% of lines and cannulae were left for more than 72 h before replacement, and significant discordance occurred between line replacement and syringe and cannula exchange. We concluded that contamination of the infusions was probably extrinsic and secondary to manipulations of the system during use. Prolonged usage and discordant exchange of infusion components were likely important factors in initial contamination and subsequent bacterial proliferation. The problem resolved following the introduction of a policy for routine and simultaneous replacement of lines and syringes at 24-h intervals and upon cannula exchange. PMID- 10363208 TI - Survival of Acinetobacter baumannii on bed rails during an outbreak and during sporadic cases. AB - Genotypic methods showed Acinetobacter baumannii biotype 9 genotype I to be the epidemic strain on an outbreak in an intensive care unit (ICU) which lasted from January to April of 1996. A cohort was established during March in which hospital personnel were assigned exclusively to A. baumannii infected or colonized patients. New patients were not admitted to the ICU until the last infected patient was discharged. However, strain I was isolated during April and vectors other than human carriage were suspected. The ICU comprised four sections; patients and beds were moved within them according the severity of diseases. Strain I was isolated from a bed rail nine days after the infected patient was discharged. This dry vector may explain the transmission of the epidemic strain between sections. The following July, four new infected patients were identified and three different strains, including the epidemic one, were recovered. The two other strains were also isolated from a bed rail. Although this environmental source does not explain by itself the transmission of an epidemic strain, it illustrates that dry vectors can be secondary reservoirs where A. baumannii can survive. PMID- 10363209 TI - Epidemiological analysis of imipenem-resistant Serratia marcescens in hospitalized patients. AB - Our objective was to examine epidemiological characteristics of hospitalized patients with imipenem-resistant Serratia marcescens. We performed a case-control study using data collected from computerized databases and chart review. Molecular typing by pulsed field gel electrophoresis of available isolates was performed. One hundred and ten patients had Serratia spp isolated during the 23 month study period. Twelve were infected or colonized with S. marcescens resistant or of intermediate susceptibility to imipenem. Eleven of the 12 patients were detected during a seven-month period between August 1994 and February 1995, suggesting the possible occurrence of an outbreak. However, the patients were admitted to different wards and services and, in eight patients, imipenem-resistant S. marcescens were isolated within 48 h or admission. None of the patients had epidemiological links within other institutions. The 12 cases were not more likely to have been exposed to beta-lactam antibiotics, including imipenem, than patients with imipenem-susceptible isolates. Six isolates were available for typing by PFGE; three were indistinguishable or closely related whereas each of the other three isolates were unique. In conclusion both the prevalence of imipenem-resistant S. marcescens and its unusual epidemiologic characteristics warrant further study. PMID- 10363210 TI - The influence of methicillin-resistant Staphylococcus aureus (MRSA) carriers in a nursery and transmission of MRSA to their households. AB - We examined two persistent MRSA-carrier nurses in a maternity hospital to elucidate the transmission of methicillin-resistant Staphylococcus aureus (MRSA) from healthcare providers to newborn infants and to the nurses' own families. Genotyping of the MRSA strains was performed by analyzing genomic DNA restriction length polymorphisms from pulsed-field gel electrophoresis (PFGE-RFLPs). The children of these nurses were carrying genotypically identical MRSA strains as their mother. Both MRSA carrier families remained asymptomatic over a two-year follow-up period. Eradication of nasal MRSA carriage from the two nurses resulted in declining MRSA carriage rates among infants in the nursery. Healthcare providers may become transient or persistent MRSA carriers whilst working in hospitals in which MRSA is endemic. They may then become a source of infection for patients as well as their own families. We recommend that healthcare providers should be examined for MRSA if an MRSA epidemic occurs in a hospital. The families of any such carriers should also be examined for MRSA. PMID- 10363211 TI - Nurses failure to appreciate the risks of infection due to needle stick accidents: a hospital based survey. AB - One of the most important occupational risks to healthcare workers is exposure is to blood-borne viruses. This study examined nurses' perceptions of risk of contracting infection following single or multiple exposure to blood or body fluids. Two hundred and ninety nurses were surveyed using a questionnaire. One hundred and thirty-three responded; 85 worked in higher risk areas (ITU, Haematology, Haemodialysis and Neonatal Surgical Units) (Group A) and 48 worked in lower risk areas (medical wards, an orthopaedic and an ENT ward) (Group B). Forty-nine percent of subjects from group A and 60% of subjects from Group B believed that a needle stick injury with a needle contaminated with infected blood was an unlikely source of infection. Fifteen percent from group A and 20% from group B thought that infection with a blood-borne virus following a needle stick injury contaminated with Human Immunodeficiency Virus (HIV) infected blood was very unlikely. Twelve percent from group A and 10% from Group B did not know whether resheathing needles between use can provide protection against HIV. Sixty seven percent from group A and 71% from group B disagreed with the statement that nurses are at higher risk of exposure to HIV/HBV than the other healthcare workers. Thirteen percent from group A and 5% from group B agreed with the statement, whereas 8% from group A and 5% from group B thought that nurses are at less risk. Only 22% from group A and 23% from group B would take more precautions if they knew that the patient had HIV/HBV infection, whilst 11% and 8% respectively admitted that they would take special precautions only when the patient has clinical symptoms of HIV/HBV infection. The findings suggest that these nurses would benefit from further education regarding infection from blood borne viruses. PMID- 10363212 TI - Correlation between surface and air counts of particles carrying aerobic bacteria in operating rooms with turbulent ventilation: an experimental study. AB - Airborne contamination with bacteria-carrying particles (cfu/m3) and their sedimentation rate (cfu/m2/h) was compared in an operating room (OR) equipped with two turbulent ventilation systems. One was a thermally based system with inlet of cool clean air at the floor level and evacuation of the air at the ceiling by convection (17 air changes/h). The other was a conventional plenum pressure system with air supply at the ceiling and evacuation at the floor level (16 air changes/h). The study was made during rigidly standardised sham operations (N = 20) performed in the same OR by the same six member team wearing non-woven disposable or cotton clothing. Airborne contamination in the wound and instrument areas was related to the surface contamination rate in the same areas and in addition, on the patient chest and in the periphery of the OR. With the exception of the periphery of the OR, the surface and air contamination rates were highly correlated in both ventilation systems (P = 0.02-0.0006, r2 = 0.52 0.79). This was also true particularly when disposable clothing was used while the correlation was weaker in cotton clothing experiments. An equation describing the relation between surface and air counts is given. Typically, the surface counts were numerically 16-fold the air counts, i.e., the number of colonies sedimenting on four 14 cm-diameter agar plates during 1 h will almost equal the number of airborne cfu per m3. We propose, that sedimentation plates represent not only a technically easier method than air sampling but when correctly used, are also the most realistic indicator of airborne bacterial OR contamination in areas critical for surgery. PMID- 10363213 TI - Swinging back the MRSA pendulum? PMID- 10363214 TI - Revised guidelines for control of MRSA in hospitals: finding the most useful point. PMID- 10363215 TI - MRSA and hospital control. PMID- 10363216 TI - First report of MRSA from hospitalized patients in Sudan. PMID- 10363217 TI - Testing water quality for automatic washer--disinfectors. PMID- 10363219 TI - Saliva ejectors in dentistry. PMID- 10363218 TI - Water quality for endoscope washer-disinfectors. PMID- 10363220 TI - Informed consent in school health research: why, how, and making it easy. AB - The obligation to obtain informed consent for student participation in health related research creates a complex set of legal, ethical, and administrative responsibilities because the interests of research integrity are delicately balanced against protection of human subjects. Even the term itself sparks a range of responses depending on one's perspective and stake in the process. This paper traces the historical impetus behind obtaining informed consent, identifies key elements comprising informed consent, and reviews types of consent procedures used in schools. The authors suggest 20 ways to boost response rates while providing a realistic level of informed consent for school-based studies. PMID- 10363221 TI - Short-term impact of safer choices: a multicomponent, school-based HIV, other STD, and pregnancy prevention program. AB - This study evaluated the effectiveness of the first year of Safer Choices, a theoretically based, multicomponent HIV, STD, and pregnancy prevention program for high school youth. The study featured a randomized trial involving 20 schools in California and Texas, with a cohort of 3,869 ninth-grade students. Students who completed both the baseline and the first follow-up survey approximately seven months later were included in the analysis (n = 3,677). Safer Choices enhanced 9 of 13 psychosocial variables including knowledge, self efficacy for condom use, normative beliefs and attitudes regarding condom use, perceived barriers to condom use, risk perceptions, and parent-child communication. Safer Choices also reduced selected risk behaviors. Specifically, Safer Choices reduced the frequency of intercourse without a condom in the three months prior to the survey, increased use of condoms at last intercourse, and increased use of selected contraceptives at last intercourse. PMID- 10363223 TI - Psychosocial factors associated with dieting behaviors among female adolescents. AB - This study determined whether female adolescents who were attempting weight loss (dieters) differ from those who were not (nondieters) with respect to a set of psychosocial factors. The sample consisted of 2,536 normal-weight and underweight female adolescents who participated in the National Longitudinal Adolescent Health Survey. Psychosocial factors examined included depression (four measures), self-esteem, trouble in school, school connectedness, family connectedness, sense of community (two measures), grades, autonomy, and protective factors. MANCOVA revealed significant differences between dieters and nondieters. Self-esteem was the strongest contributing factor differentiating dieters and nondieters. These results have implications for health education and health promotion with regard to both primary and secondary prevention. Self-esteem building should be incorporated within the parameters of a comprehensive school health program and certainly should be a component in any nutrition education program aimed at preventing unhealthy dieting behaviors. By understanding the factors associated with these behaviors, it may be easier to identify individuals attempting weight loss despite being of normal or low body weight. PMID- 10363222 TI - Mothers' perceptions of factors influencing violence in schools. AB - This study investigated mothers' perceptions of factors contributing to school violence. Of 345 mothers, 225 (65%) from urban public schools and 120 (35%) from suburban public schools, significant differences in perceptions of school violence were found on the enabling factors subscale for school location. Urban school mothers were significantly more likely than suburban mothers to attribute violence problems at their child's school to the lack of dress codes, violent messages in rap music, and poor parent/teacher communication. Significant differences in perceptions of school violence were found on the reinforcing factor subscale for school location, income, family structure, and race. Mothers of low- and middle-income, single parents, and African Americans were much more optimistic about the possibility that violence prevention programs for students, parents, and teachers would work well to stop or reduce school violence than were higher-income, married, and White mothers. These mothers also were more likely to believe it was acceptable for their child to fight at school than were their counterparts. PMID- 10363224 TI - High school health teachers' perceived self-efficacy in identifying students at risk for suicide. AB - A national random sample of 228 high school health teachers completed a 45-item survey to examine their perceived self-efficacy regarding adolescent suicide. Most respondents were female, White, and held master's degrees. Most believed it was their role to recognize students at risk for suicide, believed that if they did recognize students at risk it would reduce the chances that the student would commit suicide, and believed that one of the most important things they could do would be to prevent a suicidal student from committing suicide. However, only 9% believed they could recognize a student at risk for suicide. High efficacy expectations scores were associated with working at a school that offered an inservice program on adolescent suicide, included teaching about suicide prevention in the curriculum, and had a crisis intervention team. This study suggests that teacher health education programs should spend more time on developing the skills necessary to identify students at risk. In addition, a comprehensive school suicide prevention program is strongly encouraged for all high schools. PMID- 10363225 TI - Why don't you want to "go with" me? The dynamics of interpersonal attraction. PMID- 10363226 TI - Instinctual sadism: a recurrent myth about human nature. PMID- 10363227 TI - The myth of redemptive violence: implications for developmental theory and clinical practice. PMID- 10363228 TI - Developmental metaphors: the dialectic of perception and creation in the construction of a novel. PMID- 10363229 TI - Psychotherapy as a short story: selection and focus in brief dynamic psychotherapy. PMID- 10363230 TI - The illusion of a nonfuture: reflections on psychoanalysis in the twenty-first century. PMID- 10363231 TI - A labyrinth of connections: when the patient generates an analytic community. PMID- 10363232 TI - On the concepts of amae and the mother-group. AB - Notwithstanding Freud's (1921) early linkage of psychoanalytic group psychology with patriarchal themes (i.e., father-leader), more recent findings suggest that the regression in group behavior in general, and in group psychotherapy in particular, tends to evoke frequent maternal ideations. In this connection, this article points to a phenomenological relationship between the individual's universal need to be at one with the internalized mother, as exemplified in the Japanese concept of amae and that of the Mother-Group. With amae serving as an explanatory construct for the group members' interpersonal relations (i.e., identifications and transferences), the idea of the Mother-Group ties in with the group entity's simultaneous supportive functions (i.e., "we are all in the same boat") coupled with its role as a collective arena for the reactivation of unconscious wishes for the nurturing mother of early childhood. In linking a Japanese psychological concept with another, which was developed in the West, I am disproving Rudyard Kipling's often quoted assertion that "never the East and the West shall meet." I was encouraged in this task by Takeo Doi's written comment to me that the idea of the Mother-Group "certainly can be linked with that of Amae. For instance, when the group functions as the Mother-Group, you can say that the participants are experiencing Amae in one way or another" (T. Doi, Sept. 17, 1993, personal communication). In an extended sense, one hopes this essay might even serve to revive our interest in the interrupted well-known early collaboration of psychoanalysts and of anthropologists, which had begun to forge a way to connect personality formation with its cultural context (LeVine, 1974). The insufficient attention paid to the influence of cultural factors on attachment behavior was recently decried by Bretherton (1997). PMID- 10363233 TI - Attitudes of embeddedness and transcendence in psychoanalysis: subjectivity, self experience, and countertransference. PMID- 10363234 TI - Psychodynamic psychotherapy in an AIDS nursing home. PMID- 10363235 TI - Contingency and the unformulated countertransference: a case presentation. PMID- 10363236 TI - Men's love for men: contrasting classical American film with The Crying Game. PMID- 10363237 TI - [Clinical course of idiopathic inflammatory myopathies: complications, survival and prognostic factors]. AB - OBJECTIVES: To describe the clinical and epidemiological characteristics, complications, survival and prognostic factors of a series of patients with idiopathic inflammatory myopathy (IIM) diagnosed with homogeneous criteria in the same center. PATIENTS AND METHODS: Patients with the diagnosis of IIM during an inclusion period of 20 years were studied. They were classified following the criteria of Bohan and Peter, and Dalakas. Epidemiological, clinical and therapeutical data were obtained in all cases. Evolution and survival were analyzed with the Kaplan-Meier and Cox multiple regression models. RESULTS: One hundred thirty-five patients with IIM were included in the study: 32 polymyositis (PM), 90 dermatomyositis (DM) and 13 inclusion body myositis (IBM). Forty-six percent presented some complications attributed to the disease or treatment, and 10 with PM, 29 with DM and 3 with IBM died during the follow-up. The probabilities of survival were 86% the first year, 80% the second year, 71% the fifth year, and 57% the tenth year. Infections and cancer were the main death causes. While survival analyses did not show independent risk factors for PM, advanced age, presence of associated neoplasm, raised erythrocyte sedimentation rate (ESR) and muscle relapse were identified as a poor prognostic indicators for DM, whereas raised ESR and long lasting symptoms prior to diagnosis of the myopathy were for IBM. CONCLUSION: In spite of the therapeutic advances, IIM are still diseases with high mortality and morbidity. PMID- 10363238 TI - [Prognostic value of heterozygosity loss in chromosome 3p in non-microcytic bronchogenic carcinoma]. AB - BACKGROUND: Recent advances on carcinogenesis have led to the recognition of different patterns of behaviour of non-small cell lung cancers apart from those guided by the TNM staging system and the histologic subtype. The frequent genetic loss on chromosome 3p in all kinds of lung carcinoma leads to the suspect of the presence of a tumor suppressor gene located in that place. The aim of this work was to compare the different clinical features and evolution after treatment of the patients with non small cell lung carcinoma with and without loss of heterozygosity (LOH) on 3p. PATIENTS AND METHOD: Forty-five operated on non-small cell lung cancer patients were evaluated. The mean age was 64.4 years and all the patients were male. Seven patients had been previously treated for another epithelial neoplasm. 82.2% of the patients were heavy tobacco smokers. Most of the tumors (62.2%) were squamous cell carcinomas. Samples of tumoral and non tumoral lung tissue were immediately frozen after surgery. DNA from the tissue was extracted; then PCR amplification of microsatellites in regions 3p14 and 3p21 was performed. To determine the LOH in the regions analyzed a polyacrylamide gel electrophoresis was performed. RESULTS: Twenty-five percent of the informative samples for 3p14 and 21.9% for 3p21 showed LOH. There was an statistical relationship between the LOH on 3p14 and the history of tobacco smoking and the adenocarcinoma histologic subtype (p < 0.05). There was a higher number of relapses and a shorter disease-free interval in those patients harboring 3p21 LOH. CONCLUSIONS: LOH on 3p can be detected in non-small cell lung carcinoma. Patients with loss of heterozygosity on 3p21 have a worse evolution after a curative intended surgical resection. PMID- 10363239 TI - [Validation of the Spanish version of the Liebowitz social anxiety scale, social anxiety and distress scale and Sheehan disability inventory for the evaluation of social phobia]. AB - BACKGROUND: Social phobia is an anxiety disorder of increasing interest in clinical psychiatric practice and research. The questionnaires most widely used in the psychometric evaluation of these patients are: Liebowitz Social Anxiety Scale (LSAS), Social Anxiety and Distress Scale (SADS) and Sheehan Disability Inventory (SDI). The objective of this study was to evaluate the validity and reproducibility of the Spanish versions. SUBJECTS AND METHODS: Convergent validity was analysed by correlating patients' scores on the LSAS, SADS and SDI with scores on the Global Activity Evaluation Scale (GAES), the Hamilton Anxiety Scale (HAM-A) and the Visual Analogue Scale (VAS) of the EuroQol. Validity of the internal structure was examined by analysing the correlations between the different sub-escales of the questionnaires. Internal consistency was analysed using Cronbach's alpha and Kuder-Richardson coefficients. Discriminative capability was analysed by comparing LSAS, SADS and SDI scores of patients with social phobia with scores from healthy subjects, and with that obtained on the HAM-A and the EuroQol VAS. Reproducibility was analysed by re-testing patients after 15 days. RESULTS: 57 patients and 57 healthy subjects were recruited in 4 psychiatric centres. The three questionnaires showed an adequate convergent validity with the GAES, the HAM-A and the EuroQol VAS (r = -0.24-0.40 and r = 0.29-0.52). The LSAS and SDI questionnaires showed a homogenous internal structure in terms of correlation between sub-scales (r = -0.61-0.93 for LSAS, and r = -0.04-0.61 for SDI). All the sub-scales of the questionnaires showed an adequate internal consistency (with coefficients between 0.72 and 0.88). The questionnaires discriminated between groups of patients with different levels of symptom severity and self-perceived overall health. They also discriminated between patients with social phobia and healthy subjects (area under the Receiver's Operating Characteristics curves = 0.95-0.99). All sub-scales from the questionnaires showed adequate reproducibility (with intraclass correlation coefficients between 0.63 and 0.88). CONCLUSIONS: The Spanish versions of the LSAS, SADS and SDI questionnaires have shown adequate validity and reproducibility for use in clinical research and the clinical assessment of patients with social phobia in Spain. PMID- 10363240 TI - [Determinants associated wtih the presence of risk behaviors in HIV infected patients]. AB - BACKGROUND: Determinants associated with risk behaviours are evaluated in a known HIV-infected population not belonging to the great metropolitan nuclei. PATIENTS AND METHODS: 110 unselected HIV+ patients were interviewed, including 77 variables. Their association with sharing needles, and unprotected sex is analysed. RESULTS: Sharing needles was associated to: low academic achievement (p = 0.045), no children (p = 0.045), any physical limitation (p = 0.004), previous admission to detoxification unit (p = 0.014), and depression. With unprotected sex were associated: low academic achievement (p = 0.005), lesser time of HIV infection (p = 0.009), no family support (p = 0.005), and scanty information about HIV transmission (p = 0.018). CONCLUSIONS: A cohort of HIV-infected subjects who persist with risk practices is remaining. Some easily recognizable variables may be useful for their early recognition. PMID- 10363241 TI - [Idiopathic myositis: a disease or a syndrome?]. PMID- 10363242 TI - [Confidentiality in human rights]. PMID- 10363243 TI - [Genetic study of a new family with Hippel-Lindau type IIB disease]. AB - BACKGROUND: Von Hippel-Lindau disease is characterized by the variable presence of cerebellar and retinal haemangioblastomas, phaeocromocytomas and hypernephromas, beginning at early stages of life. Von Hippel-Lindau gene has been located in the short arm of chromosome 3 (3p25.5) and has been involved in the regulation of DNA transcription acting as a suppressor gene. More than 500 different mutations have been described. SUBJECTS AND METHODS: We describe a new family with the type IIB Von Hippel-Lindau disease in which, apart from clinical studies, we performed a genetic screening trying to identify germinal mutations. RESULTS: So far, we have point out 6 patients with the G-->A transversion at codon 167 (R167Q). Two of them with overt clinical disease (phaeocromocytoma in case II.1 and haemangioblastoma in the II.2) at the beginning of the study and one with a non-suspected clinical presentation (phaeocromocytoma and renal carcinoma in case I.1) out of 8 family members studied in three generations. CONCLUSIONS: The genetic screening in this family permitted us to identify three subjects before their clinical onset. The absence of the mutation in two of the younger patients will simplify the clinical follow-up of this family. Genetic screening must be generalized in the follow-up of Von Hippel-Lindau disease families, because of economic advantages and clinical efficacy. PMID- 10363244 TI - [Intellectual property, authorship and confidentiality in multicenter studies: the experience of MPTR. Working group of MPTR]. PMID- 10363245 TI - [Bacterial resistance to antibiotics. More than a clinical problem? Vancomycin resistant Enterococcus spp. as an example]. PMID- 10363246 TI - [Resistance to erythromycin in strains of Streptococcus pyogenes isolated in the province of Barcelona]. PMID- 10363247 TI - [The first case of beta-thalassemia by frameshift CD5 (-CT) mutation in Spain]. PMID- 10363248 TI - [Ticlopidine and hepatic adverse reactions. Data from the Spanish drug surveillance system]. PMID- 10363249 TI - [Treatment of community acquired acute uncomplicated pyelonephritis]. PMID- 10363250 TI - [Image of the week. Classic case of polyarteritis nodosa]. PMID- 10363251 TI - [Trigeminal neurinoma]. PMID- 10363252 TI - [Serial change of cerebral blood flow after collagenase induced intracerebral hemorrhage in rats]. AB - In case of intracerebral hemorrhage (ICH), many clinical studies have reported that great alterations in regional cerebral blood flow (rCBF) have been observed not only in the acute but also in the chronic stage. However, due to the absence of reproducible animal model, experimental studies have failed to confirm the evidence. We therefore studied the serial change of rCBF and investigated the possibility of secondary brain damage using our collagenase induced ICH model. A total of 60 adult male rats were examined. 2 microliters artificial cerebrospinal fluid, which was adjusted to a pH 7.4 by a buffer, containing 0.25 units of bacterial collagenase was stereotactically injected into the left caudoputamen in 28 rats. 2 microliters artificial cerebrospinal fluid without collagenase was also injected into 28 control rats under sham operation. The remaining 4 rats were studied as a non treatment group. The rCBF of the frontal cortex, parietal cortex, temporal cortex, occipital cortex, caudoputamen, thalamus and the cerebellum was determined bilaterally by [14C]-iodoantipyrine quantitative autoradiography according to Sakurada's method at 2, 4, 12, 24, 48, 168 and 720 hours after the operation. At 4 hours after injection, significant decrease of rCBF was observed in the ipsilateral hemisphere and also in some contralateral regions. In the ipsilateral frontal cortex and caudoputamen, there was significant increase of the rCBF which reached close to the baseline at 24 hours. After that, the rCBF of those regions decreased significantly again and remained at a low value even 1 month after injection. In the other regions of the ipsilateral hemisphere which were located remotedly from the hematoma, the rCBF recovered from 24 to 48 hours after injection. In the contralateral hemisphere, the rCBF also recovered at 24 hours. We suggest that this alteration of the rCBF observed in perihematomatous regions was the result of secondary brain damage caused by the existence of hematoma. On account of this, we conclude that early removal of hematoma is necessary to obtain a better result in cases of ICH. PMID- 10363253 TI - [Usefulness of galea suturing method for scalp closure]. AB - A method for scalp closure to prevent alopecia along a suture line is described. Only the galea is sutured. By tacking a sufficient width of the galea on both sides with an absorbable suture material, the sutured wound forms a ridge. The outer layer is then closed with skin staples to keep the blood circulation undisturbed. This procedure contrasts with the traditional method in which the galea is sutured with the overlying subcutaneous tissue and consequently the hair follicles are strangled. By adopting the method of suturing the galea, development of alopecia along a suture line has been effectively prevented and scarring has become less conspicuous. PMID- 10363254 TI - [Histological study of vascular structure between the dura mater and the outer membrane in chronic subdural hematoma in an adult]. AB - The authors reported the results of histological study of the vascular structure between the dura mater and the outer membrane of chronic subdural hematoma (CSH) in an adult. Many vascular connections were detected at particular part in CSH. According to the microscopical findings and location, these connective vessels were classified into 3 types. 1) Capillary-like vessels had thin walls and narrow diameters. They originated in the middle layer of the dura mater and were connected to macrocapillaries (MC) in the outer membrane. We could not perceive the flow direction in these vessels. 2) Small veins had thin walls and wide diameters. They gathered blood flow of many MC in the outer membrane and entered into the dura mater. Some of them penetrated through the total layer of the dura mater, then flowed into the paraarterial venous sinus of the middle meningeal artery (mmA). 3) Small artery had a thick wall and was about 50 mu in diameter. Though histological continuity was not proved, it was ascertained that the artery originated from a branch of the mmA, and entered the outer membrane. After entering, the small artery separated into many small branches which spread throughout the whole of the outer membrane. The vascular structure of the outer membrane consisted mainly of MC and small arteries with many branches. In this study. We were not able to find the location where both vessels of the outer membrane connected. PMID- 10363256 TI - [Glioblastoma fed by meningeal branches of the external carotid artery: a case report]. AB - A rare case of glioblastoma fed by meningeal branches of the external carotid artery was reported. A 63-year-old female was transferred to our hospital suffering from gait disturbance and dysarthria. CT and MRI revealed brain tumor and paratumoral hemorrhage with a large cyst that was heterogeneously enhanced and existed in the right fronto-temporal region. Right external carotid arteriography demonstrated the tumor stain markedly fed by the right middle meningeal artery and the accessory meningeal artery. Subtotal removal operation was carried out uneventfully using the right fronto-temporal craniotomy. The histological diagnosis was glioblastoma. After the operation the patient was in good condition, and was transferred to another hospital for the purpose of the synchronized chemoradiotherapy. It is well known that any glioma invades the meninges. However, we rarely encountered an intra-axial glioma fed by a meningeal blood supply. A meningeal-invaded glioma may make difficult its differentiation from meningioma. We concluded that there is necessity for close examination of the intra-axial brain tumors invaded and fed by meningeal blood supply. PMID- 10363255 TI - [Presurgical mapping with functional MRI: comparative study with transcranial magnetic stimulation and intraoperative mapping]. AB - PURPOSE: The accuracy of preoperative mappings in patients with brain tumors near the central sulcus using functional magnetic resonance imaging (fMRI) or transcranical magnetic stimulation (TCS) was evaluated by comparative reference to intraoperative mapping. METHODS: The thumb movement was evoked by TCS for the mapping of the motor cortex. After the placement of the marker determined by TCS on the scalp, fMRI under motor tasks consisting of repetitive grasping was performed. For motor cortex activation, an axial oblique plane to maximize gray matter sampling in the rolandic cortex was employed in order to compare these different mapping techniques more precisely. Sixteen patients with brain tumors were included in this study. RESULTS: In nine patients, fMRI disclosed activation in one restricted gyrus or in the localized area around one restricted sulcus. Of these nine patients, preoperative TCS mapping corresponded closely with fMRI in six, while in the remaining three, the TCS marker fell between 1 and 2 cm apart from the fMRI-activated area. However, in these three patients, intraoperative electrocortical stimulation corresponded with the preoperative mapping with fMRI. In six patients, contigucus two gyri were activated by motor tasks. The TCS marker was disclosed on one of the two activated gyri. Of these six patients, the position of the TCS marker and fMRI-activated site corresponded with each other in four cases. They were found on the same gyrus but there was 1.0-2.0 cm distance between them in two cases. Intraoperative somatosensory evoked potential was monitored in two of these six cases. They corresponded well with the mapping by fMRI and TCS together. In only one patient, no significant activation area was obtained by fMRI because of excessive head motion during motor tasks. The TCS marker in this patient was identical with intraoperative electro-cortical stimulation mapping. CONCLUSION: For presurgical planning in patients with brain tumor near the central sulcus, comparative evaluation with fMRI and TCS is applicable and provides accurate functional mapping. PMID- 10363257 TI - [Two cases of an intracranial large hemangiopericytoma associated with brain swelling in the early operative stage]. AB - Intracranial hemangiopericytomas are hypervascular tumors. Consequently, resection of these tumors must occasionally be stopped owing to profuse bleeding. We report two cases of large intracranial hemangiopericytoma whose resection had to be stopped owing not to bleeding but to brain swelling. These patients were treated with large doses of barbiturate until the intracranial pressure had normalized. After preoperative irradiation therapy, the residual tumors were easily removed without serious complications. We speculate that the brain tissue around a large hemangiopericytoma might be under ischemia because of long standing compression and deviation of blood to the hypervascular tumor. A large amount of blood flowing into these brain tissues due to external and internal decompression inverts cerebral perfusion acutely from ischemia to hyperemia. The brain swelling in the early operative stage would be evoked by this "inverted cerebral hyperemia". We found that brain swelling is sometimes encountered during the resection of a large hemangiopericytoma even in the early operative stage. If swelling occurs, tumor resection should be stopped immediately and intracranial pressure should be controlled. The residual tumor can then be easily removed without complications after preoperative irradiation therapy. PMID- 10363258 TI - [A case of primary cerebral neuroblastoma surviving for eight years]. AB - We report a case of a patient with primary cerebral neuroblastoma who has survived for 8 years. A 10-year-old boy was admitted to our hospital because of headache and nausea. CT scan on admission revealed a large cystic tumor on the right frontal lobe. Subtotal tumor resection was carried out. A second operation was performed for the residual tumors which were removed meticulously with confirmation of the absence of tumor cells on each frozen section. After tumor removal, YAG laser was applied at each local area. Histological diagnosis disclosed primary cerebral neuroblastoma. Because of postsurgical meningitis and parent's refusal, neither chemotherapy nor radiation therapy was performed. There have been no findings of the tumor recurrence during the last eight years, and now the patient is enjoying high school life to the full, without any neurological deficits. In reviewing the literature, outcomes of neuroblastoma cases are very poor. Our case seems to be one of the rare long-survival cases. PMID- 10363259 TI - [Intracerebral hemorrhage associated with long-lasting deficiency of factor XIII]. AB - A 5-year-old boy had an intracerebral hematoma evacuated from the left parieto occipital region at a local hospital. There was a difficulty in hemostasis during the operation. Postoperatively his neurological state gradually deteriorated. CT scans showed a gradual increase in intracerebral hemorrhage at the operated site. He was transferred to our clinic for further treatment. The preoperative blood coagulation profile showed that the activity of factor XIII was only 4%. The hematoma was removed without any trouble under administration of factor XIII. Postoperative course was also uneventful although the plasma level of factor XIII did not go up higher than 26% in spite of its daily administration. Although factor XIII administration was terminated one month after cranioplasty, the patient showed good recovery and the level of factor XIII spontaneously normalized in 6 months. The cause of the extremely low level of factor XIII in this case is not known, but it could be ascribed to massive hemorrhage. PMID- 10363260 TI - [Recanalization of cerebral cortical venous thrombosis: a case report]. AB - Several clinical cases of sinus thrombosis have been reported, but localized cerebral cortical venous thrombosis is a fairly rare clinical entity. We report a case of a 51-year-old woman who presented with right hemiparesis on admission. CT scan revealed subcortical hemorrhage with perifocal edema in the left front parietal lobe. A T2W image revealed a large area of hyperintensity indicating edema, and coronal Gd enhanced T1 weighted image showed a reverse-triangle-shaped enhanced lesion. Left carotid angiogram showed cerebral cortical venous thrombosis in the left frontal lobe, but the superior sagittal sinus was not occluded. A month after admission, right hemiparesis had suddenly improved and the patient could walk without any support. Left carotid angiogram showed recanalization of thrombosed cortical veins and CT scan showed disappearance of any low density lesion indicative of edema. It is an important fact that neurological deficit improved quickly a month after onset, and it was suspected that, even without resorting to fibrinolytic therapy, recanalization of thrombosed cerebral veins would still eventually occur. We found the reverse triangle-shaped enhanced lesion on MRI and we suspected that it was typical finding for cerebral cortical venous thrombosis. PMID- 10363261 TI - [Endodermal cyst of the cerebellopontine angle cistern: case report]. AB - We report a case of a 55-year-old man with endodermal cyst located in the cerebellopontine angle cistern. The patient presented with dizziness. Magnetic resonance imaging (MRI) revealed multilocular cystic lesion at the right cerebellopontine angle. T1-weighted image showed a mass with a low signal intensity, but higher intensity than CSF. Gd-DTPA T1-weighted image showed no enhancement in the mass. Diffusion-weighted image showed a mass with no signal lesion. After successful surgical removal, it was found to be an endodermal cyst. These cysts have usually been found in the spinal canal, and their intracranial occurrence is exceptional. The unusual location of the cyst and its histological features and radiological findings are discussed. PMID- 10363262 TI - [Rendu-Osler-Weber disease with cerebral hemorrhage due to a capillary telangiectasia: a case report]. AB - We encountered a case of acute cerebral hemorrhage secondary to capillary telangiectasia in a 10-year-old female. She was diagnosed as having Rendu-Osler Weber disease (ROW). In this case, the cerebral hematoma did not result in neurologic damage and the final outcome was excellent. ROW is an autosomal dominant disorder characterized by the presence of vascular malformations of varying types in several tissues, including the brain, nasal mucosa, lungs, gastrointestinal tract, and liver. Neurological complications occur in 8 to 12 percent of patients with ROW. The pons is the most common site of capillary telangiectasia, but most of the malformations caused are clinically silent. Massive cerebral hematoma due to capillary telangiectasia is rare. Cerebral hematoma due to hypertension in a child is less than that found in an adult. So in a child it is important to investigate the origin of cerebral hematoma. PMID- 10363263 TI - [Newly identified peroxisomal disorders]. PMID- 10363264 TI - [Striatonigral degeneration and sporadic olivopontocerebellar atrophy: a consideration of the clinical entity of multiple system atrophy]. AB - Striatonigral degeneration (SND) and sporadic olivopontocerebellar atrophy (sOPCA) are categorized under multiple system atrophy (MSA), since these disorders have common clinical and pathological features. However, it is still uncertain whether these disorders are manifestation of the same disease. In this study, we performed both clinical and neuroradiological studies on patients with SND or sOPCA in whom clinical diagnosis was based on a criteria during eight years in our hospital. A total of forty patients had SND and thirty-one patients had sOPCA. The onset ages of patients with SND (60.7 +/- 8.7, mean +/- SD) were significantly higher than those with sOPCA (55.4 +/- 7.9). In both SND and sOPCA patients, about 20% had clinical symptoms suggesting the involvement of multiple systems: pyramidal, cerebellar, extrapyramidal and autonomic symptoms. In 55% of the SND patients, cerebellar symptoms could be observed, and the same percentage of sOPCA patients had parkinsonism. Although, as defined, cerebellar symptoms were predominant in sOPCA patients and parkinsonism was predominant in SND patients, the SND patient group was particularly homogeneous with respect to clinical characteristics. The initial symptoms of SND were parkinsonian gait or tremors. Almost all patients exhibited asymmetrical appearance of the parkinsonian symptoms, such as rigidity, tremors, and bradykinesia. Tremors at rest were observed in two-thirds of the patients with SND during the course of their illness, but dementia was infrequently observed. There was no detectable limitation in horizontal eye movements in patients with SND. The progression of the disability of patients with SND was rapid; according to the clinical rating scale of parkinsonism, the average level of disability deteriorated to Hoehn Yahr's stage III after three years from disease onset, and then deteriorated to stage IV after four years. Neuroradiologically, only a small proportion of patients with SND (27%) showed magnetic resonance image (MRI) findings suggesting OPCA pathology, such as volume loss in the brainstem or cerebellum with/without T2-high signaling of transverse fibers of pons or T2-high signaling of the middle cerebellar peduncle. Simultaneously, a small proportion of the patients with sOPCA (20%) showed MRI findings suggesting putaminal pathology, such as T2-low intensity signals of the putamen with linear T2-high intensity signals around the lateral putamen. Our results suggest that SND and sOPCA can be clearly differentiated, at least from clinical or neuroradiological aspects. Since there is still no evidence indicating that each disorder is a clinical variant of a single disease caused by the same etiology, a differentiation might be important for future pathogenetical studies. PMID- 10363265 TI - [Superiority of intrafascicular neurography over conventional nerve conduction studies in evaluating axonal degeneration]. AB - We investigated conventional motor and sensory nerve conduction studies (MCS & SCS) with regard to the sensitivity in detecting axonopathies. Twelve patients with axonal type of polyneuropathy, 2 vincristin neuropathy and 10 cisplatin neuropathy, were examined by MCS & SCS. Their data were compared with those by intrafascicular microneurography (MNG) of the median nerve. Nerve conduction velocities were within normal limits or slightly reduced to 87-99% of the normal. Amplitude of compound muscle action potential (CMAP) by MCS decreased to 4 or 5 mV in vincristin neuropathy, though cisplatin neuropathy presented normal amplitude. Amplitude of sensory nerve action potential (SNAP) by SCS was undetected in one median nerve and in three sural nerves. While, compound nerve action potential (CNAP) by MNG was all recorded, and presented the amplitude value of below 150 microV in seven patients. The waveform was normal or mild neuropathic pattern. No patients presented normal CNAP amplitude and reduced SNAP amplitude. But there were three patients who had normal SNAP amplitude and reduced CNAP amplitude. In SCS we could recognize abnormal only after CNAP amplitude by MNG dropped to below 100 microV. Cisplatin neuropathy demonstrates reduction of CNAP or SNAP amplitude, and vincristin neuropathy further presents reduction of CMAP amplitude. Evaluation of axonopathy is best achieved by nerve action potential amplitude. Conventional surface electrode methods are available for this purpose, but MNG is more sensitive and is capable of quantitative analysis even in severely damaged nerves. PMID- 10363266 TI - [A clinical study on long-term outcome in patients with syringomyelia associated with Chiari malformation]. AB - To investigate the long-term outcome in patients with syringomyelia associated with Chiari malformation (chronic tonsilar herniation), we investigated the actual factors of the patient's problems by a questionnaire for the patient. Replies to the questionnaires were obtained from 44 patients with syringomyelia who had been treated with expansive suboccipital cranioplasty with dural plasty (with plugging of the central canal in 20 patients). In 25 cases (68%) in whom the motor dysfunction of the upper extremities remained, the symptoms improved in 16 cases (43%) and were unchanged in 9 cases (24%). In 18 cases (64%) in whom the motor dysfunction of the lower extremities remained, the symptoms improved in 10 cases (36%) and were unchanged in 8 cases (29%). In 31 cases (78%) in whom the sensory disturbance remained, the symptoms improved in 19 cases (48%) and were unchanged in 12 cases (30%). The motor dysfunction of the upper extremities persisted significantly in more patients having a duration of illness over 2 years than in patients with those less than 2 years. Sensory disturbance persisted significantly in more patients with a duration of illness over 3 years than in patients with that less than 3 years. These results suggest that firstly, sensory disturbance (disturbance of the posterior horn) is most apt to remain, then motor dysfunction of the upper extremities (disturbance of the anterior horn) and followed by motor dysfunction of the lower extremities (disturbance of the pyramidal tract). We conclude that patients should be treated before having irreversible spinal cord disturbance. PMID- 10363267 TI - [The relationship between coagulation time and bilateral occurrence in chronic subdural hematoma]. AB - The relationship between the bilateral occurrence and coagulation time was analyzed in 137 cases of chronic subdural hematoma. Bilateral chronic subdural hematomas more frequently happened in the patient with coagulation time elongation beyond the normal control. This tendency was significant in the non traumatic group and bilateral subdural hematomas were found in: 45% in prothrombin time (PT)-elongated group vs 10.3% in non-elongated group and 38.9% in activated partial thromboplastin time (aPTT)-elongated group vs 14.6% in non elongated group. The ratio of PT and aPTT coagulation time against the normal control was significantly larger in the bilateral group than in the unilateral group. The ratio was as follows: PT ratio was 1.056 in bilateral group and 0.995 in unilateral group. aPTT ratio was 1.051 in bilateral group and 0.954 in unilateral group. Furthermore the bilateral hematomas tended to happen in hemodialysis cases, and in patients on warfarin or anti-platelet drugs. Bilateral hematomas were found in 41.7%, 37.5% and 33.3% respectively. The ratio of PT and aPTT coagulation time was as follows: PT ratio was 1.040 in hemodialysis cases and 1.082 in warfarin-applicated cases. aPTT ratio was 1.022 in hemodialysis cases and 1.055 in warfarin-applicated cases. These results suggest that the suppressed coagulation ability and platelet function are involved in the genesis of bilateral chronic subdural hematomas. PMID- 10363268 TI - [Location of primary somatosensory area in cerebral arteriovenous malformation involving sensorimotor area]. AB - The purpose of this study was to investigate the cortical reorganization associated with congenital brain lesion such as intracerebral arteriovenous malformation (AVM). Dipole source localization of somatosensory evoked potential (SEP) was performed in five patients with AVM encompassing sensorimotor cortex. Dipole tracing method combined with the scalp-skull-brain head model (Homma et al., 1994) was used to locate dipole source of an early cortical component of SEP elicited by median nerve electrical stimulation. The location of dipole source of SEP, which could be considered as the hand area of primary somatosensory area, was shown in the realistic section of the head and could be easily superimposed on the magnetic resonance imaging. SEP was recorded three times in each patients and the results were reproducible. In 2 patients whose postcentral gyrus was not involved in AVM, the dipole source of SEP was localized in the intact postcentral gyrus. The locations of dipole sources of SEP in the both hemisphere were symmetrical. In 3 patients whose postcentral gyrus was encompassed by AVM, the dipole source of SEP was localized in the surrounding intact gyrus which was distant from the usual region of postcentral gyrus. Somatotopy was different from the normal pattern. The hand area was located more medially than usual observed in normal postcentral gyrus. Despite the paucity of the number of patients and data obtained by dipole source localization, our findings support the existence of reorganization in the cerebral cortex with congenital lesion such as AVM. These findings of aberrant mapping of cortical function may be explained by the plasticity of brain function. The developing brain can inherit function that would normally have been performed by the region of brain involved in the lesion. We demonstrated that dipole tracing of SEP was a noninvasive method used to localize areas of eloquent cortex in patients harboring AVM. This method is of value in treatment planning. PMID- 10363269 TI - [Vein of Galen aneurysmal malformation: multi-staged feeder clipping in two neonatal cases with intractable heart failure]. AB - The authors reported the surgical experience of two cases of vein of Galen aneurysmal malformation in the newborn, whose congestive high-output cardiac failure was intractable. Along with the intensive care to clinical manifestations of the heart failure, multi-staged feeder clipping was carried out to decrease the high-flow shunt of the malformation. As stages going on, heart failure was relieved gradually and cathecolamines were weaned. Although certain retardation became apparent in both cases, they are showing satisfactory development in the long-term follow up. By the recent advancement of the embolization technique, the embolization appears to have already taken place the treatment of choice for this malformation. According to the neonatal evaluation score of Lasjaunias, the embolization would no longer be recommendation in neonates, whose general condition scored less than eight points. The authors believe, based on our two cases, that multi-staged feeder clipping is one of the effective modality of treatment in neonates of the vein of Galen aneurysmal malformation with severe multiorgan failure. PMID- 10363270 TI - [Clinical and neuroradiological features of spontaneous intracranial hypotension: report of two cases]. AB - We report two patients with spontaneous intracranial hypotension (SIH) showing bilateral subdural hematoma. One of the two patients was a 32-year-old woman, and the other was a 27-year-old healthy woman. Both patients presented chronic, intractable, orthostatic headache with dizziness and nausea. In both patients, both general and neurological examinations were normal, and routine laboratory tests were all normal, except for dry taps of the lumbar puncture. Brain CT scans and MRI revealed thin, bilateral subdural hematomas. RI-cisternography and CT myelography disclosed multiple extraspinal CSF leakages along the nerve root sheathes of the cervical segments and early bladder filling of the radionucleotides in both patients. These findings support an emerging hypothesis that the extraspinal CSF leakage may play a role for inducing SIH. Anatomical fragility around the nerve sheath, especially that of the lower cervical segments, may contribute to the pathophysiological mechanism underlying SIH. For making a prompt diagnosis of SIH and for the better understanding of the pathophysiological mechanism of SIH, RI-cisternography of the whole spinal segments is important. PMID- 10363271 TI - [A case of primary Sjogren's syndrome presenting with aseptic meningoencephalitis associated with sarcoidosis]. AB - A 64-year-old Japanese woman with sarcoidosis and primary Sjogren's syndrome was admitted to our hospital in January 1998 for consciousness loss. Physical examination revealed meningeal signs and hyperreflexia in the lower limbs. Laboratory investigations showed hypergammaglobulinemia and high titers of autoantibodies such as RF, ANA, anti SS-A and SS-B antibody. In addition, she had keratoconjunctivitis, and an apple-tree pattern was observed in sialography, but serum ACE was normal. In the cerebrospinal fluid (CSF), the cell count, total protein, and interleukin 6 (IL-6) were increased, but ACE in CSF was normal. We diagnosed the case as primary Sjogren's syndrome presenting with aseptic meningoencephalitis. After treatment with prednisolone, her manifestations and CSF findings including IL-6 resolved. Therefore, the level of IL-6 in CSF may be an indicator for the activity of primary Sjogren's syndrome associated with central nervous manifestations. PMID- 10363272 TI - [Metastatic skull tumors from cancers associated with subcutaneous mass lesions]. AB - The incidence of metastatic brain tumors is increasing because of the recent progress in the detection and management of primary cancer. However, metastatic skull tumors from cancers associated with giant subcutaneous mass lesions are rare. We present four patients with metastatic skull tumors: two from hepatic cancer, one from lung cancer, and one from mamma cancer. In these patients, plain skull X-ray and bone CT showed osteolytic lesions. Angiograms revealed a tumor stain fed by abnormal vessels from the external carotid artery. MRI demonstrated masses with marked homogeneous enhancement with the "dural tail sign" in the dura adjacent to the tumors in three skull tumors from hepatic and mamma cancers, and a mass with slightly enhancement without the "dural tail sign" in a skull tumor from lung cancer. At surgery, hemorrhagic well-demarcated tumors were totally removed. The histological diagnosis was skull metastases from cancers in all cases. In cases with the "dural tail sign" on MRI, no tumor cells were seen in the inner layer of the dura and the dura adjacent to the tumors. It is possible that the "dural tail" is due to increased vascular permeability of the dural vessels. The recurrence of these skull tumors was not observed during the follow up period. Surgical treatment for the metastatic skull tumors from cancers may be indicated to prevent deteriorating neurological symptoms affecting the quality of life. PMID- 10363273 TI - [A case of cerebral vascular malformation with hemiatrophy and paroxysmal dyskinesia]. PMID- 10363274 TI - [Brachial plexus neuropathy (brachial plexopathy)]. PMID- 10363275 TI - [A case of nasopharyngeal carcinoma presenting with various neurological deficits in the course of treatment]. PMID- 10363276 TI - Molecular karyotype diversity in the microsporidian Encephalitozoon cuniculi. AB - The microsporidian Encephalitozoon cuniculi can infect numerous mammals, including man. Three strains of E. cuniculi have been identified so far, the major marker being the number of a tetranucleotide repeats in the rDNA internal transcribed spacer. We investigated diversity at the chromosomal level through the electrophoretic karyotypes obtained from 15 E. cuniculi isolates from 5 different host species. All preparations provided patterns with 9-12 bands within a narrow molecular size range. Six karyotype forms were distinguished, involving subdivision of strain I into 3 types (A, B, C) and strain II into 2 types (D, E). The types A, B and C were mainly associated with isolates from rabbits of different geographical origins. The types D, E and F were characterized by a reduced chromosome size range, 2 of these appearing specific to a carnivorous host species (D in dog and F in blue fox). Hybridization experiments showed that all E. cuniculi isolates possess 11 chromosomes, with a size polymorphism entailing occasional electrophoretic comigration of heterologous chromosomes and differential migration of homologous ones. DNA rearrangements should occur during mitosis and the hypothesis of diploidy for the basic state of E. cuniculi seems likely. PMID- 10363278 TI - Occurrence of Leishmania major in sandfly urine. AB - Promastigotes of Leishmania major were frequently detected in the urine droplets discharged by infected Phlebotomus papatasi and P. duboscqui females during feeding. Parasites were present in the urine of 37.5% P. papatasi and 16.1% P. duboscqi females, even in those with low intensity gut infections. Free-swimming forms (elongated nectomonads, short slender promastigotes and metacyclic forms) predominated in excreted droplets. Viability of excreted parasites was proved by cultivation on blood agar, and the presence of metacyclic forms in urine droplets was confirmed by specific fluorescence assay with 3F12 antibodies. While the release of promatigotes from the anus of the sandfly was frequent, these were rarely egested from the mouth-parts of sandfly females (1.3% for P. duboscqi and 0% for P. papatasi) fed on microcapillaries, even if the females were heavily infected. The possible role and significance of the discharge of parasites in sandfly urine are discussed. PMID- 10363277 TI - High prevalence of Enterocytozoon bieneusi in swine with four genotypes that differ from those identified in humans. AB - The microsporidial species Enterocytozoon bieneusi is found among immunocompromised, particularly HIV-infected, patients with chronic diarrhoea, and rarely also among immunocompetent persons with self-limited diarrhoea. Only recently, E. bieneusi was detected in 4 pigs in Switzerland raising the question of a potential zoonotic nature of this parasite. We examined faecal samples of 109 pigs, 24 cows, horses and red foxes each for the presence of E. bieneusi by PCR and compared these isolates with isolates obtained from stool samples of 13 HIV-infected patients living in Switzerland. In animals, E. bieneusi was only identified in pigs with a prevalence of 35%. Analysis of the rDNA internal transcribed spacer (ITS) sequence allowed the classification of E. bieneusi from 28 pigs into 4 distinct genotypes which grouped very closely (identity 96.3 98.8%) together with 2 of the 3 human-derived E. bieneusi genotypes. Hence, E. bieneusi seems to be a common parasite in swine, but no genotypes were identified that were found in humans. Nevertheless, swine might serve as a new animal model for enterocytozoonosis. PMID- 10363279 TI - Optimization of conditions for growth of wild-type and genetically transformed Trypanosoma cruzi on agarose plates. AB - Growth of Trypanosoma cruzi as colonies on solid medium has not been widely used as an experimental procedure. We therefore sought to establish a reliable and routine plating method. The optimal results were achieved with a matrix of 0.65% low melting point agarose onto which epimasigotes from the mid-to-late logarithmic phase of growth were spread. Colonies could be isolated after incubation for 21 days in a humidified 5% CO2 environment at 28 degrees C. Plating efficiencies in the range of 40% were obtained by this method and clones could be recovered into liquid medium or onto blood-agar slopes with a high success rate. The procedure has also been adapted for the isolation of genetically transformed clones after electroporation of epimastigotes with either plasmid or cosmid vectors. This was best achieved by inclusion of the electroporated cell inoculum in a 0.6% agarose overlay containing G418 as the selective drug, on top of a 0.8% agar base. Transformation efficiencies were as high as 10(-5) cells per microgram of DNA. A reliable plating method for T. cruzi will have many applications and is a significant step towards the use of 'shotgun transformation' to generate libraries of T. cruzi recombinants. PMID- 10363280 TI - Trypanosoma brucei spp. development in the tsetse fly: characterization of the post-mesocyclic stages in the foregut and proboscis. AB - Post-mesocyclic development of Trypanosoma brucei in the tsetse fly in its migration from midgut to salivary glands, was revisited by sequential microdissection, morphometry and DNA-cytofluorometry. This development started by day 6 after the infective feed, with passage of mesocyclic midgut trypomastigotes through proventriculus and upward migration along foregut and proboscis to the salivary gland ducts. Kinetics of salivary gland infection showed that colonization of the salivary glands by epimastigotes occurred only during the time-limited presence of this developmental phase in the foregut and proboscis. Post-mesocyclic trypanosomes in the foregut and proboscis were pleomorphic, with 4 morphological stages in various constant proportions and present all through from proventriculus up to the salivary gland ducts: 67% long trypomastigotes, 27% long epimastigotes, 4% long epimastigotes undergoing asymmetric cell division and 2% short epimastigotes. Measurements of DNA content demonstrated a predominant tetraploidy for 67% of these trypanosomes, the remainder consisting of the homogeneous diploid short epimastigotes and some long epimastigotes. According to the experimental data, the following sequence of trypanosome differentiation in the foregut and proboscis is proposed as the most obvious hypothesis. Incoming mesocyclic trypomastigotes (2N) from the ectoperitrophic anterior midgut start to replicate DNA to a 4N level, are arrested at this point, and differentiate into the long epimastigote (4N) which give rise, by an asymmetric cell division, to 2 unequal, diploid daughter cells: a long, probably dead-end long epimastigote and a short epimastigote. The latter is responsible for the epimastigote colonization of the salivary glands if launched at the vicinity of the gland epithelium by the asymmetric dividing epimastigote. PMID- 10363281 TI - Development of monogenean communities on the gills of roach fry (Rutilus rutilus). AB - The formation and development of monogenean communities on the gills of roach fry was followed in 1992 from early June to October (size range 9 to 47 mm). Roach fry (n = 291) were sampled weekly from the small, humic River Rutajoki in central Finland. A further 209 roach fry were reared in a fish farm supplied by water from the river. Four Dactylogyrus species were found: D. nanus, D. crucifer, D. micracanthus and D. suecicus. Other species found on the gills were Gyrodactylus sp. and Paradiplozoon homoion. The first Dactylogyrus juvenile occurred on a 12 mm long fish fry in late June and the first adult (D. nanus) 1 week later in Tank 1. D. nanus was also the most common parasite in the river. Young fry had high numbers of Gyrodactylus sp. on the gills compared with adult roach in previous studies. Signs of site preference were found; D. nanus and D. micracanthus preferred the 2nd gill-arch and Dactylogyrus juveniles the outmost one. Abundances of monogenean infracommunities increased until the middle of August when, on average, 1.9 species and 3.7 worms per fry were noted and decreased then as the water temperatures fell. A lack of competition between species is suggested; for example, D. crucifer appeared on the gills at that time when abundance of D. nanus was at its highest. The roach fry hatched and reared in the fish farm revealed that larvae of the most common dactylogyrid species were able to disseminate and colonize over 120 m from the river to the farm. Influence of change colonization events was shown by the appearance and development of Gyrodactylus sp. population on roach fry in only 1 of the 2 tanks at the farm. PMID- 10363282 TI - Role of acetylcholinesterase (AChE) secreted by parasitic nematodes on the growth of the cell line from epithelial origin HT29-D4. AB - The excretory-secretory (E-S) products of the parasitic nematodes Trichostrongylus colubriformis and Nematodirus battus were found to modify the in vitro proliferation of the tumorous colic HT29-D4 cell line of epithelial origin. A characteristic feature of these E-S products is the presence of a high level of acetylcholinesterase (AChE) activity, the biological significance of which remains unclear. To determine a possible role of AChE on cell growth, the enzyme was purified from E-S products using edrophonium chloride. Purity was confirmed by polyacrylamide gel electrophoresis, using silver and Karnovsky stains, before assessing its effects on cell proliferation. The purified AChE was incorporated at different concentrations in a culture medium of HT29-D4 cells. A mitogenic effect was shown for low concentrations (0.1-14 units). By contrast, an inhibitory effect was noted at high concentrations (35-1400 units). Furthermore, polyclonal antibodies were prepared and depletion of AChE in E-S products by immunoprecipitation or affinity chromatography resulted in a partial or total disappearance of the stimulatory effect of cell growth. Thus, the results form this in vitro study suggest a modulatory role for AChE secreted by nematode parasites on the proliferation of epithelial cells of the host. PMID- 10363283 TI - Are there temporarily non-infectious dauer stages in entomopathogenic nematode populations: a test of the phased infectivity hypothesis. AB - Many studies of entomopathogenic nematodes (Heterorhabditidae and Steinernematidae) have reported that only a small proportion (typically < 40%) of infective stages (dauers), even under apparently ideal conditions, actually infect a host. The 'phased infectivity hypothesis' is most frequently invoked to explain this pattern of low infection with entomopathogenic nematodes. It proposes that at a given point in time not all individuals are infectious i.e. infectiousness is delayed in some individuals. We tested experimentally several predictions based on this hypothesis. Specifically, if phased infectivity occurs, we should be able to expose dauers to increasing numbers of potential hosts until dauers no longer infect and still be able to recover viable dauers. These recovered dauers which did not infect should be infectious at some later point in time. However, our results do not support the phased infectivity hypothesis for 3 species of Steinernema: most dauers could be recovered in one sampling round when provided with sufficient suitable hosts. In contrast, Heterorhabditis bacteriophora frequently did not infect all available hosts, and infectious dauers were recovered in subsequent sampling rounds. This result is more consistent with the phased infectivity hypotheses, but further research is needed before we can be more confident in the hypothesis. For all species tested, the number of available hosts influenced population levels of nematode infectivity. This suggests that the infection status of hosts can influence whether a dauer infects. Our results indicate that phased infectivity is not a common phenomenon in entomopathogenic nematode dauers, despite the widespread acceptance of this hypothesis. PMID- 10363284 TI - The surface coat of infective larvae of Trichinella spiralis. AB - The surface coat of the infective larvae of the parasitic nematode Trichinella spiralis was characterized with respect to its biophysical properties, morphology and composition. Labelling of larvae with the fluorescent surface probe PKH26 was lost after activation (by incubation in mammalian medium containing trypsin and bile), or following pronase treatment. Electron microscopical examination revealed that pronase treatment resulted in the loss of an amorphous surface layer only, further demonstrating the specificity of PKH26 for the larval surface coat. Surface coat shedding was inhibited by sodium azide and carbonyl cyanide, or by incubation of larvae at 4 degrees C, suggesting the shedding process required metabolic energy. Pre-labelled, unactivated larvae demonstrated continuous slow surface coat shedding and could be re-labelled with PKH26, indicating that the shed coat is replaced in these parasites. However, pre labelled larvae which were activated failed to re-label with the probe, suggesting that activation provides an irreversible trigger for surface changes. PKH26, therefore, is a useful marker for larval activation. Examination of the shed coat material by scanning electron microscopy revealed 2 types of morphologies; one comprising thin multilaminate sheets and the other of amorphous material with ridges producing a fingerprint-like motif. Western- and lectin blotting of the shed coat material demonstrated 2 prominent entities; a 90 kDa glycoprotein, which bound Datura stramonium agglutinin and was resistant to N- and O-glycanase treatment and a 47-60 kDa set of protein(s). Analysis of the surface lipids by electrospray mass spectometry revealed the presence of lysophosphatidic acid (lysoPA, C14:2) and an unidentifiable component of 339.4 Da. These two lipids constituted 36.9% and 36% by mass of surface coat lipids respectively. The presence of lysoPA was confirmed by thin layer chromatography, which also detected phosphatidic acid (PA). The polar lipids detected in solvent rinses of intact parasites by electrospray mass spectrometry were PI (C48:4), PE (C40:4 and C38:4), PS (C40:4), lysoPC (C20:2 and C18:2) and lysoPA (C14:2). These observations are discussed with respect to the role of the surface coat and its shedding in the T. spiralis host-parasite relationship. PMID- 10363286 TI - [Comparative clinico-radiological study of lacunar infarcts of the cerebral hemispheres and brain stem in 110 cases]. AB - INTRODUCTION: Lacunar infarcts (LI) are small deep infarcts due to occlusion of perforating branches. OBJECTIVE: Our objective was to outline the clinical and epidemiological characteristics which differentiate hemispherical lacunar infarcts (HLI) from those of the brain stem (SLI). PATIENTS AND METHODS: We present 110 cases of LI (80 HLI, 30 SLI) analysing risk factors, clinical syndromes, findings on neurological examination (dysarthria, gravity, distribution and proportional paresia), form of clinical presentation, evolution whilst in hospital, site and results of carotid duplex. Diagnosis was made in 72 patients using magnetic resonance (MR) and in 38 patients using computerized axial tomography (CT). RESULTS: The commonest characteristics of SLI, as compared with HLI, with statistical significance (p < 0.05) was the appearance of supranuclear facial paresia (OR = 2.68), severe motor involvement (OR = 4.23), form of presentation with previous TIA (OR = 6.33), fluctuating evolution of the symptoms (OR = 5.78) and progression of the paresia (OR = 6.41). Also, in the pontine LI there was significant correlation between site and gravity: the lower the site of the lesion, the more serious was the paresia. Patients with multiple LI presented with no previous risk factors significantly more frequently than those with a single LI. CONCLUSION: The different clinical profiles may help to establish the subgroups of IL, according to where they occur. PMID- 10363285 TI - Genetic variation and epidemiology of Echinococcus granulosus in Argentina. AB - Polymerase chain reaction-ribosomal ITS-1 DNA (rDNA) restriction fragment length polymorphism (PCR-RFLP) analysis and sequencing of the mitochondrial cytochrome c oxidase subunit 1 (CO1) and NADH dehydrogenase 1 (ND1) genes were used to characterize 33 Echinococcus granulosus isolates collected from different regions and hosts in Argentina, and to determine which genotypes occurred in humans with cystic hydatid disease. The results of the study demonstrated the presence of at least 4 distinct genotypes; the common sheep strain (G1) in sheep from Chubut Province and in humans from Rio Negro Province, the Tasmanian sheep strain (G2) in sheep and 1 human from Tucuman Province, the pig strain (G7) in pigs from Santa Fe Province and the carnel strain (G6) in humans from Rio Negro and Buenos Aires Provinces. The finding that pigs harboured the pig strain and the occurrence of the Tasmanian sheep strain has considerable implications for the implementation of hydatid control programmes due to the shorter maturation time of both strains in dogs compared with the common sheep strain. Furthermore, this is the first report of the presence of the G2 and G6 genotypes in humans which may also have important consequences for human health. PMID- 10363287 TI - [A proposal for application and scoring of the Clock Drawing Test in Alzheimer's disease]. AB - INTRODUCTION: The Clock Drawing Test (CDT) has been used in recent years as a simple neuropsychological instrument to assess cognitive deterioration associated with dementia, even though uniform operative criteria with respect to its application and scoring have not been established. OBJECTIVE: To present application normatives and establish the most relevant psychometric criteria of the CDT in a sample of healthy subjects (HS) and patients with Alzheimer's disease (AD). PATIENTS AND METHODS: 56 patients were selected of which 35 were female and 21 were male. The patients' mean age was 72.7 with a standard deviation of 7.64. All of whom where probable AD patients according to the NINCDS ADRDA criteria at stage 1 CDR. The group of HS was made up of 56 control subjects (34 female, 22 male) with a mean age of 72.14 and a standard deviation of 7.2. The CDT was applied in both its command (COM) and copy (COP) experimental conditions. RESULTS: The main psychometric parameters analysed in the studied series showed the following values: internal consistency (Cronbach's alpha coefficient 0.9029); cut off point CT COM 6 with 92.80, sensitivity; false negatives (FN) 7.2 with a specificity rating 93.48; false positive (FP) 6.52 with 93.16 efficacy; cut off point CT COP 8 with 73.11 sensitivity; FN 26.89 with 90.58 specificity; FP 9.42 with 82.49 efficacy. CONCLUSION: The CDT can be used to discriminate between HS and those in the initial stages of AD in the given sample using the established application and scoring criteria. PMID- 10363288 TI - [Hyperbaric oxygenation in subcortical frontal syndrome due to small artery disorders with leukoaraiosis]. AB - INTRODUCTION: Frontal leukoaraiosis (LA) is a common finding in patients with subcortical small-vessel disease and currently its pathogenesis is attributed to ischemic-hypoxic mechanisms. It associates to a vascular subcortical frontal syndrome (VSFS) for which an effective treatment does not exist. CLINICAL CASES: We present four subjects from a prospective patient-blind controlled pilot trial to study efficacy and safety of hyperbaric oxygen therapy (HBO) vs hyperbaric air in VSFS with LA. All of them had frontal or extended LA on computed tomography scan and lacunes in basal ganglia and centrum ovale, with moderate to severe gait disorders, urinary dysfunction, cognitive impairment, and dependence in the daily living activities. Deficits had begun two to ten years before and had remained stable three months previous to the treatment. Patients were assessed with validated scales and tests one week before and after being administrated ten daily sessions of HBO at 2.5 atmospheres absolute for 45 minutes with a multiplace chamber. Serious adverse effects did not occur. After treatment a noticeable gait, urinary and cognitive improvement was observed in all subjects, increasing their independence. They remained clinically improved during four to five months, after which the previous deficits reappeared. Then, three patients received ten daily sessions of air at 1.1 atmospheres absolute for 45 minutes (controls) and the other a new HBO regimen, which improved as the first time. From the controls, there were no changes in two, while the other did only improve cognitively. CONCLUSION: These patients show that HBO is effective and safety in reversing, at least partially, although at great length, chronic neurological deficits associated to vascular frontal LA, highlighting that a functional reserve therapeutically useful exists. PMID- 10363289 TI - [Discrimination between genetic factors in attention deficit]. AB - INTRODUCTION AND OBJECTIVE: In order to elucidate the genetic and environmental components involved in the susceptibility to develop attention deficit hyperactivity disorder (ADHD), a complex segregation analysis on nuclear families (n = 53) ascertained from affected probands belonging to Medellin, in the Antioquian State, Colombia, was performed. METHODS AND RESULTS: Models of cohort effect (non-inheritance), multifactorial, recessive major gene, non-major gene component and non-transmission of major gene were rejected. Contrarily, dominant and codominant major gene models and non-multifactorial component could not be rejected. Thus, the better model fitting the data was that of the major gene (dominant/codominant). This major gene explains more than 99.99% of the ADHD phenotypic variance (value of heritability in the mixed model equal to 0.007%), which permit to assume a low aport of the environmental component to the phenotype ADHD. Gene frequency of the major gene was 3% in the general population of Antioquia and its penetrance was closed to 30%. CONCLUSION: Some cautions and aspects related to the bias of the interview and diagnosis of the parents are discussed. PMID- 10363290 TI - [Evoked potentials in the sacred baboon: long-term follow-up of intracerebroventricular infusion of nerve growth factor]. AB - INTRODUCTION AND OBJECTIVE: It is well known that in aged animals cognitive deficit occurs, homologous with that occurring in Alzheimer's disease in humans, and as has been shown in others species, this may be attenuated by administration of nerve growth factor (NGF). Therefore the basic aim of this study was to make an electrophysiological evaluation of the repercussion that there might be after long-term administration of this neurotropin in the sacred baboon (Papio hamadryas) comparing aged with young animals. MATERIAL AND METHODS: We studied a six year old male and a 39 year old female, after sedation. Long-term intraventricular administration of NGF was carried out using a continuous infusion pump, at a dose of 2.1 micrograms/kg/day. Recordings were made before installing the pump and 1, 3 and 6 months after insertion. A Neuropack Four-mini set for evoked potentials (Nihon Kohden) was used to record auditory evoked potentials from the brain stem and visual evoked potentials due to flash. RESULTS AND CONCLUSION: In both animals there were modifications of their electrophysiological responses. These reached a maximum after one month, more markedly in the older animal and this could possibly be related to the neuromodulator effect of NGF. PMID- 10363291 TI - [Benign partial convulsions in adolescence]. AB - INTRODUCTION: Loiseau and Orgogozo first described benign partial convulsions of adolescence (BPCA). OBJECTIVE: To analyze the clinical and electroencephalographic characteristics of adolescent patients diagnosed as having BPCA. PATIENTS AND METHODS: We reviewed the clinical histories of 125 patients with onset of partial epilepsy between the ages of 10 and 18 years, followed-up at the J.P. Garrahan Hospital, Buenos Aires, between 1990 and 1997. Fourteen of these patients fulfilled diagnostic criteria for BPCA. We analyzed the following clinical and electroencephalographic parameters: sex, personal and family history, age of onset, semiology, distribution, duration and frequency of crises, neurological examination, neuro-images (brain CT/MR) using conventional techniques, time of follow-up and evolution. RESULTS: Ten patients were boys (71.4%) and four girls (28.6%). The follow-up period was from one to seven years (average 3.1 years). Convulsions had started at between 10 and 16 years old, average 12 years old, with a peak of maximum incidence at between 12 and 13 years old. The crises were of partial simple type in 12 patients (85.7%) and partial complex type in 2 (14.3%); nine cases had secondarily generalized tonic-clonic convulsions (64.2%). Crises were brief in 100%, in 13 cases (93%) they occurred whilst awake and in one case (7%) during sleep. CONCLUSIONS: BPCA occur in a transient condition which affects males more often than females, has a peak onset at between 12 and 13 years of age and is characterized by simple motor partial crises, frequently with secondary generalization, single or inclusters, during waking and is of a benign course. Neurological examination, EEG between crises and neuro-radiological studies were normal. PMID- 10363292 TI - [A system of multidimensional behavior assessment. A scale for parents of children from 6 to 11 years of age. Colombian version]. AB - INTRODUCTION: Behavioral Assessment System for Children (BASC) has demonstrated to be useful in the diagnosis of Attention Deficit Disorder (ADD). PATIENTS AND METHODS: A randomized sample of 120 children, 6 to 11-year-old, participants from the school of the city of Medellin, Colombia, was selected. The sample was stratified by sex and two socioeconomic status (SES). Parents were asked to answer the BASC Parent Rating Scale (PRS) 6-11, authorized Spanish version. RESULTS: Cronbach's alpha coefficient was 0.85 for the clinical scale (9 items). It was 0.75 for the Adaptive Scale (3 items). A scale designed with 4 items to assess ADD (hyperactivity, attention problems, aggression, and conduct problems) showed an alpha coefficient of 0.82. Male children scored significantly higher than female (ANOVA, p < 0.05) in hyperactivity, conduct problems, and atypicality. Children from low SES scored significantly higher than children of high SES on the most of clinical measures (p < 0.05) and lower on the three adaptive measures. Cluster analysis selecting six clusters found a prevalence of 61.6% for normal male children. In the total sample there were a 4% at risk of DDA type II (inattentive) and 14% at risk of DDA type I (combined). CONCLUSIONS: BASC PRS (6-11) showed reliability and validity to assessing the behavior in Spanish speaking Colombian children. PMID- 10363293 TI - [A study of the motor and sensory cortex using functional magnetic resonance: tasks of active and passive movement]. AB - INTRODUCTION AND OBJECTIVE: The objective of this study was to locate the rolandic area (pre- and post-central) by means of functional magnetic resonance imaging (FMRI) and define its correspondence on a Talairach map, whilst active and passive movements of the dominant hand were performed. MATERIAL AND METHODS: Ten healthy volunteers were found, 6 men and 4 women, of an average age of 26 years (range 22-33). Two appropriate tasks were designed: one involving active and one passive movement. The examination was carried out using a 1.5 Tesla (General Electric) MRI apparatus. An echo-sequence of planar echo-gradient (BOLD technique) was used, making sagittal and axial planes, parallel to the AC-PC line (anterior commissure-posterior commissure). Subsequently an anatomofunctional Talairach map was drawn for each subject, to include the information obtained on FMRI. RESULTS: In all subjects central activity was detected in the rolandic area during the tasks involving selected active and passive movements. Overlap was seen between the pre- and post-rolandic areas with both types of tasks. CONCLUSION: There is good correlation between the image obtained of motor-sensory activity in the rolandic zone and the Talairach anatomofunctional map. PMID- 10363295 TI - [Dysphasia and dyslexia in the light of PASS theory]. AB - INTRODUCTION: The PASS theory of intelligence understands the cognitive function as an information process or program that can be differentiated in planning, attention, successive and simultaneous. Every process is linked to an anatomical region: planning to frontal cortex, attention to frontal cortex and subcortical structures, successive to frontal cortex and non-frontal cortex and simultaneous to non-frontal cortex. Effective remediation is possible when a PASS pattern is known. OBJECTIVE: To verify whether dysphasia and dyslexia have a typical PASS pattern. PATIENTS AND METHODS: The subjects were divided in three groups comprising 12 children with dysphasia, 12 children with dyslexia and 10 children with dysphasia and dyslexia, all of them between 6 years and 12 years of age, the majority boys. All children were administered the DN:CAS test to define the PASS pattern. A control group with 45 normal children was used. RESULTS: Dysphasia shows poor successive and simultaneous process. Dyslexia shows poor successive process. The successive deficiency is poorer and in different way in dyslexia than in dysphasia which is inferable from comparative analysis between groups. CONCLUSIONS: Dysphasia and dyslexia show a typical PASS pattern that allows an appropriate remedial training as a neurocognitive approach. The PASS diagnosis is a psychogenetic diagnosis which is different from the usual diagnosis based on semiology or results obtained with tests that explore non-PASS cognitive function. PMID- 10363294 TI - [Cerebral magnetic resonance in the study of West syndrome]. AB - INTRODUCTION AND OBJECTIVE: West's syndrome (WS) is an epileptic encephalopathy of the first year of life, associated with different aetiologies. MRI of the brain allows precise determination of the type and extent of the lesions. The aetiology must be recognised in order to establish the prognosis and a suitable therapeutic approach. The objective of this study is to analyse the aetiologies of a population of children with WS and compare the results of cases diagnosed before and after using MRI. PATIENTS AND METHODS: We analyzed the clinical histories of 448 patients fulfilling the diagnostic criteria for WS (infantile spasms and a EEG with a pattern of hypsarrhythmia), 217 in pre-MRI era (group 1) and 231 in the post-MRI era (group 2). The following parameters were analyzed: type of WS, sex, duration of follow-up, age of onset of infantile spasms and particularly the neuroradiological studies. RESULTS: Group 1: symptomatic WS, 157 patients; cryptogenic WS, 60 patients. Group 2: symptomatic WS, 169 patients; cryptogenic WS, 62 patients. The aetiologies of symptomatic WS were: cortical dysplasias, neurocutaneous disorders, cerebral malformation and prenatal clastic lesions, hypoxic ischaemia, post-infection, metabolic, tumors, Down's syndrome, others and unknown cause. CONCLUSIONS: It is known that cerebral MRI gives better definition of these types of cerebral lesions than cerebral CT does. We emphasize the importance of MRI in patients with symptomatic WS for precise determination of the aetiology, and speculate as to whether some of the 21 cases of unknown aetiology of group 1 could have been diagnosed if studied nowadays. PMID- 10363296 TI - [Depression and Parkinson disease]. AB - OBJECTIVE: Depression occurs more often in Parkinson's disease (PD) than in other chronic illnesses with important disability. The relationship between the depression level and some clinical features of PD remains controversial. Frequency of depression in these patients has been estimated and relationship between this symptom with some clinical features of PD. METHODS: A diagnosis of PD was taken according to the United Kingdom Parkinson's Disease Society Brain Bank criteria. Depression status was rated with Geriatric Depression Scale (GDS). RESULTS: Sixty-two patients (56%), 24 male and 38 female, were depressed at the time of study. The frequency of depression was higher in female (61% vs 39%, p < 0.05) and younger patients with a significant difference (p < 0.001). 53.4% of the patients became depressed previous of beginning PD symptoms, being 71% female (p < 0.05). Patients with depression had had PD longer than patients without depression (7.7 vs 5.3 years old, respectively; p < 0.05). Patients with depression were found to be more affected on motor rating scales (p < 0.01). CONCLUSIONS: Depression was found in 56% of PD patients, with female predominance (61%). Frequency of depression was higher in younger patients. Depression was associated with duration of PD and an inverse relationship between depression and cognitive status was found. PMID- 10363297 TI - [Top of the basilar artery syndrome: clinico-radiological aspects of 25 patients]. AB - INTRODUCTION, PATIENTS AND METHODS: We studied 25 patients, 16 women and 9 men (average age 64.8 years; SD 8.6; range from 36 to 79 years) admitted to our hospital with clinical findings compatible with obstruction at the level of the bifurcation of the basilar artery. The patients were selected on clinical and neuroradiological criteria. RESULTS AND CONCLUSIONS: All patients included in the study had presented with two or more infarcts in the vertebro-basilar territory, related to pathology of the rostral region of the basilar artery. Diagnosis was confirmed on CT and MRI. Infarcts were found in the thalamus, brain-stem, cerebellum and parieto-occipital lobe. Infarct of the thalamus associated with another infarct in a different region was the most frequent lesion. The CT-MRI findings in the 25 cases were: 14 patients had unilateral thalamic infarcts associated with another infarct. There were 4 patients with bilateral thalamic infarcts and 5 with bilateral occipital infarcts. In 11 patients the occipital infarct was associated with another infarct at a different level, and 6 patients each had a cerebellar infarct together with a brain-stem infarct. In 12 patients the lesions were localized to three or more areas. The commonest clinical manifestations were: motor deficit (68%), abnormal eye movements (48%), cerebellar dysfunction (40%), alteration of the level of consciousness (32%), visual field defects (20%), pupil anomalies (16%). The risk factors most frequently associated were: arterial hypertension (64%), a history of cerebrovascular accident (28%), smoking (28%), diabetes mellitus (24%) and atrial fibrillation (20%). Mortality was 8%. Unlike the classical descriptions, motor deficit was the most frequent symptom in our series. PMID- 10363298 TI - [Neuro-cognitive condition of 8 year old children with congenital hypothyroidism treated early]. AB - INTRODUCTION: Congenital hypothyroidism is the most important cause of preventable mental retardation. Since the use of programmes for early detection, mental retardation due to cretinism has been abolished and current investigations seek to detect subtle damage due to foetal hypothyroidism and strategies to minimize such deficits. OBJECTIVE: This study presents the results of neurocognitive assessment of a group of 19 children aged 8 in the Cuban Programme for the early detection of Congenital Hypothyroidism. PATIENTS AND METHODS: The programme is based on the use of umbilical cord serum levels of thyrotropin (TSH), using the Elisa ultramicro method (SUMA) and substitutive treatment with sodium levothyroxine and a programme of neuropsychiatric stimulation until 5 years old. RESULTS: The results showed that the intelligence quotients of children with hypothyroidism were not statistically different from paired controls, and thus demonstrate the efficacy of the programme. On evaluation using computerized techniques for the detection of subtle disorders of the maintenance of attention, deficiencies were found. Analysis of the conventional EEG showed that in 13 cases recordings were normal, 4 had generalized cortical damage, 1 had cortico-subcortical damage and the remainder had signs of focal irritation. On the quantitative EEG, correlation between the attention variables and absolute values (MV2/Hz) were calculated for each derivation and associations found mainly in the right parietal and left frontal regions. CONCLUSION: The effect of stimulating development in these patients is discussed. PMID- 10363299 TI - [Hyperbaric oxygenation: a promising treatment for Binswanger's disease]. PMID- 10363300 TI - [The neuro-radiologist's approach to the diagnosis and treatment of diseases of the nervous system]. PMID- 10363301 TI - [Myotonic dystrophy: DNA instability in monozygotic twins]. AB - INTRODUCTION: Myotonic dystrophy is an autosomal dominant hereditary disorder with variable expression, associated with expansion of the CTG triplet on the gene which codifies myotonia situated on chromosome 19q. We present an unusual case of myotonic dystrophy in a pair of monozygotic twin sisters, determination of CTG expansion in lymphocytes of members of their family and in their father's spermatozoids. CLINICAL CASE: The patients presented the phenomenon of anticipation of paternal transmission with an expansion of triplet CTG of lymphocyte DNA in a range of 300-1,400 identical repetitions in both. DNA of the paternal lymphocytes and spermatozoids showed a similar expansion of 75 repetitions. CONCLUSIONS: CTG expansion is not due to previous expansion of DNA in the paternal gametes but to instability of DNA in the cellular mitoses following formation of the zygote. PMID- 10363302 TI - [Central neurocytoma: analysis of three cases and review of the literature]. AB - INTRODUCTION AND CLINICAL CASES: Three cases of central neurocytoma, confirmed by immunohistochemical study are reported. The central neurocytoma has recently been added to the differential diagnosis of intraventricular tumors. It is more frequent than previously thought, with high incidence in young patients. The positivity for synaptophysin and neuron specific enolase, the negativity for neurofilament protein and glial fibrillary acid protein, and the finding of elements of neuronal differentiation on electron microscopy, are the main pathological features of these tumors. CONCLUSION: The surgical treatment is the election, and radiotherapy is reserved for malignant cases or recurrence. PMID- 10363303 TI - [Polymyositis in childhood]. AB - INTRODUCTION AND OBJECTIVE: Idiopathic inflammatory myopathies are very rare in infancy. We present five cases of polymyositis in children in which the clinical variability and difficulty in diagnosis that occurs with this disorder are clearly seen, and analyze their response to steroid treatment. CLINICAL CASES: We can distinguish two groups of patients: the first is formed of the case of a two and a half month old baby with generalized hypotonia; and the second includes the other four cases, children of between 2 and 8 years old with weakness, which was mainly proximal. One of the second group later developed juvenile chronic arthritis. In all cases there was a raised CPK and a myopathic EMG, with or without spontaneous muscle activity. Muscle biopsy showed inflammatory myopathy. The first group corresponds to so-called infantile polymyositis which is characterized by generalized hypotonia. The second group includes older children in whom the clinical features of the disorder are similar to those in adults. The association of other autoimmune diseases with infantile polymyositis is exceptional. All cases show more or less improvement with corticosteroids. CONCLUSIONS: The polymyositis are extremely rare before puberty and have a broad clinical spectrum. Congenital cases have been described in infants and in older children. The biopsy results are usually unexpected. Treatment with corticosteroids leads to clinical improvement which may be only partial and is less favorable than in the dermatomyositis. PMID- 10363304 TI - [Migrainous infarct in pregnancy]. AB - INTRODUCTION AND CLINICAL CASE: We present the case of a migrainous patient who had a cerebral infarct during a migrainous crisis. She was 26 weeks pregnant. The infarct, detected on MRI was in the right thalamic region. It presented as left hemiparesia and left hemi-hypo-estesia. Laboratory tests were normal. There was full recovery from the episode. CONCLUSIONS: Migraine is considered to be a risk factor per se for stroke, especially in young women. The association of migrainous ictus, which is a diagnosis by exclusion of other aetiologies, and pregnancy is rare, as is apparent on review of the subject. In the Western world, pregnancy is not considered to be a risk factor for ictus. The functional prognosis of migrainous stroke is good, with minimal risk of relapse. PMID- 10363305 TI - [Rol of thallium-201/gallium-67 cerebral tomogammography in the differential diagnosis of cerebral space occupying lesions]. AB - INTRODUCTION: Central nervous system (CNS) neoplasms are 10% of all tumors. A metastasis of an unknown primary neoplasm should be suspected in an adult with a cerebral tumor. In this location, the origin of most of metastases (62%) is lung, breast, skin and kidney. However, a differentiation of CNS focal infection and brain tumor, based on clinical status and morphologic imaging, may be difficult. A positive Tl-201 next to a negative Ga-67 SPECT brain scans is entirely in accord with brain metastatic tumor. CLINICAL CASE: A 72-year-old man, with history of excised bladder cancer, was admitted for neurological symptoms associated with a left occipital mass demonstrated by cranial CT and brain MRI. Clinicoradiological findings suggested a neoplastic process. Two cerebral biopsies just showed inflammatory cells. Tl-201 and Ga-67 SPECT brain scans were performed and their findings, an abnormal uptake of Tl-201 in the left occipital cortex and a negative Ga-67 scan, favored a neoplastic process. Radical exeresis of the lesion showed a metastatic adenosquamous carcinoma of probably lung origin. CONCLUSION: Tl-201 in addition to Ga-67 brain SPECT scans are a valuable tool for differential diagnosis between cerebral infection and brain tumour in patients with a sole cerebral mass lesion, especially when clinicoradiological findings and biopsy results are conflicting. PMID- 10363306 TI - [Multiple progressive occlusions of intracranial arteries]. AB - OBJECTIVES: We present a case of multiple progressive occlusions of intracranial arteries, a variety of Taveras' syndrome, without smoke spirals, which may be confused with other multifocal disorders. PATIENT: A 30 year old woman was admitted to hospital with a clinical picture of fluctuating paresia of her left limbs, blurred vision and urgency of micturition which partially recovered. On examination there was paresia of the left arm, generalized increased reflexes and facial asymmetry. RESULTS: On MRI there were areas of marked hyperintensity at T2. Some of these took up gadolinium at T1 and had a serpiginous pattern, compatible with vasculopathy. On angiography, stenosis and filling defect were seen in the left carotid artery. The anterior cerebral and left Sylvian arteries were filled by the vertebrobasilar system via the posterior communicating artery and an anomalous trigeminal artery. CONCLUSIONS: The clinical picture of multiple progressive occlusions of intracranial arteries is a variety of moya-moya disease, without the typical smoke spirals, which may lead to confusion with clinical pictures of arteritis and demyelinating disorders if angiography is not done, since this is essential for diagnosis. PMID- 10363307 TI - [Reflex anoxic cerebral crises in children]. AB - OBJECTIVE: Reflex anoxic cerebral crises are due to depression of nerve function caused by a vagotonic state or vagal hypersensitivity. In this paper we propose to review the physiopathology, clinical features and diagnostic procedures of these crises. DEVELOPMENT: There are three types of reflex anoxic cerebral crises: asphyxiating anoxic crises, ischemic anoxic crises and asphyxiating ischemic anoxic crises. They are caused by sudden activation of the so-called syncopal reflex. They are characterized by diverse clinical manifestations and may often be mistaken for epileptic crises. They are related to various precipitating factors such as a blow, often to the head, an emotional stimulus or something which upsets or molests, hyperthermia, abdominal pain, headache, physical exertion, prolonged standing-up, exposure to heat, the sight of blood, feeling of fear and other factors. CONCLUSIONS: Important elements for the diagnosis of these crises are the presence of a family history of vagal crises, the presence of precipitating factors, brief duration of the crises (seconds) in most patients, and clinical signs of facial or perioral pallor or cyanosis. The test involving pressure on the eyeball and the tilting table are useful for diagnosis. However, they should be used sensibly. PMID- 10363308 TI - [The problem of epilepsy of late onset]. AB - INTRODUCTION: We have reviewed and analyzed aspects of the concept of late onset epilepsy and its medical significance. DEVELOPMENT: Analysis of the historical progression of the concept and its aetiologies show variations depending on series, geographical region and historical period in which they were carried out. Thus in former times it was considered to be synonymous with a tumorous or traumatic aetiology and more recently there is a predominance of vascular causes. CONCLUSION: Use of the different means of diagnosis for establishing the aetiology of this patient's group is given further consideration. PMID- 10363309 TI - [Magnetic resonance findings in hypertrophic degeneration of the olivary nucleus]. PMID- 10363310 TI - [Post-traumatic anosmia: findings with magnetic resonance]. PMID- 10363311 TI - [Association of parkinsonism, dementia and amyotrophic lateral sclerosis as infrequent finding]. PMID- 10363312 TI - [Intrathecal humoral immune response in pediatric patients with meningoencephalitis due to Coxsackie B5]. AB - INTRODUCTION: Childhood is sensibly affected by viral meningoencephalitis outbreaks. OBJECTIVE: To study the intrathecal humoral immune response in a group of children suffering from Coxsackie B5 meningoencephalitis outbreak. Patients and methods. Forty eight sick children were studied. Serum and cerebrospinal IgA, IgM, IgG and albumin were quantified by radial immunodiffusion. It had been evaluated by reibergrams. RESULTS: Seventeen children has blood-cerebrospinal fluid barrier dysfunction. Four different patterns of intrathecal immune humoral response were observed mainly IgG and three major immunoglobulins class. Mean cell counts was 624 +/- 517 x 10(6) cells/l with a lymphocyte predominance. CONCLUSION: An intrathecal humoral response were reported as an early patterns like in delayed non-diagnostic puncture and in evolutive puncture in adults patterns with viral meningoencephalitis. PMID- 10363313 TI - [Vigabatrin and alterations of the visual field]. AB - INTRODUCTION: Since 1997 when Ecke et al described peripheral constriction of the visual field in three patients taking vigabatrin (VGB), and persistence of the changes after the drug was stopped, study of the visual fields of persons taking this antiepileptic drug has increased. OBJECTIVE: To evaluate the ocular repercussions of monotherapy with carbamazepine (CBZ), valproate (VPA) and VGB in children with epilepsy. PATIENTS AND METHODS: We made a blind retrospective study to evaluate visual side effects using Goldmann's campimetry and direct ophthalmoscopy in 9 children treated for 2.1 years with CBZ, 12 children treated with VPA for 3.1 years and 12 children treated for 2.1 years with VGB, all as monotherapy. In recent years visual evoked potentials (VEP) have also been assessed before starting treatment and every six months during treatment. RESULTS: One child treated with CBZ had changes in vision (blurred vision which disappeared on reducing the dose), 3 treated with VGB (diplopia which disappeared spontaneously in 2 and after reducing the dose in 1) and none of the group treated with VPA had these symptoms. All the campimetry done was found to be normal. Two of the 12 children treated with VGB had reduction in the amplitude of VEP, which became normal later. CONCLUSIONS: Since the changes in vision caused by VGB seem to be irreversible and asymptomatic in most cases, campimetric study is advisable in patients taking VGB, both when starting treatment and whilst this is being continued. PMID- 10363314 TI - [Mathematical model of the dual action of G protein on the Ca2+ current in identified neurons of the snail Helix aspersa]. AB - INTRODUCTION: The effect of the dopamine on the calcium current of identified cells of the snail Helix aspersa consists on an initial decrease, followed by a subsequent increase when the drug is removed. It had been previously demonstrated that this effect is mediated by a G protein, supposing that the decrease of the current would be mediated by the G alpha subunit, while the increase would be produced by the G beta gamma subunit. OBJECTIVE: A mathematical model has been developed with the object to test if the hypothesis of the dual action of the G protein could explain the experimental results. MATERIAL AND METHODS: It has been recording by means of the 'patch-clamp' (whole cell) in identified cells of snail. On the other hand, it has been developed a mathematical model of the calcium current using a Hodgkin-Huxley model, and a simulation of the action of the G protein on this current. RESULTS: Adjusting the kinetic parameters of the channel by means of the experimental data, it has reproduced in a faithful way the behavior of the calcium current. The simulation of the action of dopamine reproduces the decrease and increase of the current by means of the serial action of the G protein's subunits. CONCLUSION: Although a mathematical model cannot demonstrate the logic necessity of the hypotheses on that is based, it is unequivocally demonstrated that the hypothesis is fully compatible with experimental results. PMID- 10363315 TI - [Vascular malformations of the central nervous system in children]. AB - OBJECTIVE: To present the experience of vascular malformations (VM) of the central nervous system (CNS) in children obtained in our centre. Since these lesions do not often present clinically in childhood, there are few series described in the literature. PATIENTS AND METHODS: We made a retrospective review of the clinical histories of patients with VM of the CNS shown on MR, angiography and/or morbid anatomy, who were aged 15 years or under. Patients with Sturge Weber or Von Hippel-Lindau syndromes were excluded. RESULTS: We describe 32 patients with VM of the CNS. Twenty four had arteriovenous malformations (AVM), 5 had cavernous malformations (CM), 2 had malformations of Galeno's vein (MGV) and one patient had a congenital fusiform arterial aneurysm. Twenty two children (68%) presented clinically with hemorrhages. This was the form of presentation in 20 patients with AVM. In 4 of the 5 patients with CM the first sign was epileptic crisis. The two patients with MGV presented with macrocephaly, in one neonate it was associated with heart failure and in on older baby with hydrocephaly. The patient with a congenital arterial aneurysm presented with paroxystic episodes of facial pain. CONCLUSIONS: The AVM are the VM which most frequently cause symptoms in childhood. PMID- 10363316 TI - [Profile of the patient who drops out of neuropediatric follow-up]. AB - INTRODUCTION: Patients who drop out of programmed follow-up cause adverse effects to both themselves and the running of the medical. OBJECTIVE: To obtain information which would permit us to modify guidelines for medical care in cases with particular diagnoses, so as to avoid interrupting follow-up. PATIENTS AND METHODS: We selected all the patients who attended during 1995, and on 31 December 1995 had not been discharged or died. We considered a patient to have dropped out of follow-up when there was failure to attend the centre during two consecutive years without a justifiable reason. Logistic regression analysis was used to determine the profile of the patients who dropped out of follow-up. RESULTS: Of the patients attended during one year and whose treatment should have been continued, 16% dropped out; 70% of those who dropped out of their follow-up treatment were somewhat older than the rest and came in particular from two of the five health districts of the province. They had tics, tension headaches and migraine. Only three independent factors are associated with dropping out: absence of complementary tests, which is favorable, and epilepsy or previous long term follow-up which is unfavorable. CONCLUSION: Reduction in the programmed follow-up of patients with conditions which do not require complementary tests may reduce the drop out rate and improve the functioning of the Neuropaediatric Clinic. PMID- 10363317 TI - [First spontaneous epileptic crisis in childhood: risk of relapse and prognostic factors]. AB - INTRODUCTION: Knowing the risk of relapse after a first epileptic crisis is important because of its repercussion on the patient. The prognosis is uncertain in some types of crisis. OBJECTIVE: To find the probability of relapse and risk factors after a first spontaneous epileptic convulsive crisis, focal or generalized, occurring in childhood or adolescence. PATIENTS AND METHODS: We followed a cohort of 139 patients from the Health District of Hospital General of Albacete who had had this type of crisis before the age of 14. Patients with acute symptomatic, febrile or neonatal convulsions were excluded. The probability of relapse was calculated using Kaplan-Meier curves, and multivariate analysis was carried out. RESULTS: The probability of relapse 24 months after the first epileptic crisis is 70% and after 76 months is 75%. Of a series of variables analyzed, the only one associated with increased risk of relapse was the 'aetiology': the relative risk of relapse in patients with previous symptomatic aetiology is 2.1 as compared with those of idiopathic aetiology (p < 0.001). CONCLUSIONS: Patients who have had a first focal or generalized convulsive epileptic crisis in childhood have a 75% probability of developing epilepsy over the subsequent 6 years; this risk is greater in those with a previous symptomatic aetiology, so this should be taken into account when deciding on the treatment of these patients. PMID- 10363318 TI - [Subjective sleep quality and spectral analysis of electroencephalography during nocturnal sleep]. AB - INTRODUCTION: The relation between subjective sleep quality and the stages of polysomnography is unclear. Computerized sleep analysis provides a reliable tool for the study of sleep dynamics. OBJECTIVE: To study the relation between subjective sleep quality and spectral EEG parameters during nocturnal sleep. MATERIAL AND METHODS: Nocturnal sleep were recorded ambulatory in 26 subjects aged from 8 to 63. The following variables were extracted from EEG signal (Fz-Pz) quantified through the Fast Fourier Transformation (FFT): delta maximum, spectral peak, delta sleep time, efficiency and desynchronized sleep. A sleep diary was administered in the morning to assess the subjective sleep quality. Multiple regression and Anova were used for statistical analysis. RESULTS: Efficiency and delta maximum became significant predictors accounting for 54% of subjective sleep variance. The item 'how was your sleep?' was answered in a different way according to the delta maximum. CONCLUSIONS: Sleep depth, measured by spectral analysis, is an important predictor of the subjective sleep quality. The sleep classification in discrete stages ignores this feature. PMID- 10363319 TI - [Mode of inheritance of idiopathic generalized non-myoclonic epilepsy in families investigated by studying members with idiopathic epilepsy with tonic-clonic crises on waking. Antioquia, Colombia]. AB - In attempt to identify the possible role of mayor genes, multifactorial inheritance, and cohort effects in the susceptibility to idiopathic epilepsy with generalized tonic clonic seizures of the awakening type (GTCS), complex segregation analysis was performed in 196 nuclear families ascertained through affected with probands with idiopathic epilepsy with GTCS belonging to the Paisa community of Antioquia (Colombia). Models postulating no transmission, single mayor locus (dominant and recessive) only, and multifactorial component only, were rejected. The models postulating no polygenic component to transmission, and no transmission of the major effect were also rejected. Thus far, complex segregation analysis suggested that a major autosomal codominant allele together with a multifactorial component (mixed model) best explains clustering of idiopathic epilepsy with GTCS in families of the Paisa community. The deficit of transmission of heterozygotes (0.17) is compatible with the existence of epistasis acting on a major gene whose frequency was estimated to be 0.0211. Its transmission variance accounts for 81% of the susceptibility to idiopathic epilepsy with GTCS. The complementary variance (19%) is due to polygenic component. PMID- 10363320 TI - [Inflammatory demyelinating chronic polyneuropathy: a contribution to the characterization of the disease]. AB - INTRODUCTION AND OBJECTIVE: The chronic inflammatory demyelinating polyneuropathy (CIDP) is a controversial sickness. No doubt it is still a diagnosis without a strict nosologic delimitation until we acquire a greater knowledge about the underlying mechanisms, or whether a specific marker is found. We evaluated its clinical and laboratory findings, to contribute to its characterization in our environment. PATIENTS AND METHODS: We reviewed the records of 37 patients diagnosed with CIDP between 1986 and 1997. They were grouped in as to sex, age, beginning age (BA), evolution time, race, clinical form, symptoms, signs and evolutive profile. We analyzed the cytochemic and immunologic studies of cerebrospinal fluid (CSF) as well as sural nerve biopsy. RESULTS AND CONCLUSIONS: We demonstrated a predominance of BA between the fifth and sixth decades, and the presentation of a mixed polyradiculoneuropathy with motor predominance and a chronic progressive evolution. It was positively correlated with protein concentration levels in CSF, elevated in the 73.5% of the patients. There was an increase in the permeability of the blood brain barrier (BBB) in 50.8%, IgG intrathecal synthesis in 5.08 and oligoclonal bands in 8.8%. We found lost of myelin sheats in 90.6% of sural nerves and onion-bulbs formation in 60.5%, which demonstrated the high sensitivity of this study to confirm the diagnosis of demyelinating process as essential substrate in the illness. PMID- 10363322 TI - [Cerebral gliomatosis with development of multifocal glioblastoma]. AB - INTRODUCTION: Cerebral gliomatosis (CG) is a diffuse infiltrating glial neoplasia which may affect any part of the central nervous system (CNS). Its diffuse infiltrating growth leads to difficulty with clinical suspicion and imaging technique diagnosis. Magnetic resonance (MR) is more sensitive than computerized tomography (CT) for the detection of lesions. However, the extent of the infiltration may be roughly evaluated using current imaging techniques. OBJECTIVE: In this article we describe histological aspects of this rare condition, its biological behavior and correlation with radiological findings, and review the contribution of other techniques (positron emission tomography, immunohistochemical examination) in its diagnosis and delimitation. CLINICAL CASE: We present a case of CG in a 26 year old man. On CT no alterations were seen. On MR there was diffuse involvement of the white matter extending to the cortex. The patient worsened rapidly and later developed two focal masses of glioblastoma multiform in areas with the most neoplastic infiltration. CONCLUSIONS: MR is more useful than CT in establishing the diagnosis and extent of CG. Although it is a rare condition, it should be included in the differential diagnosis of conditions which affect the white matter in a diffuse manner. Poor delimitation between white and grey matter helps in diagnosis of this condition. PMID- 10363321 TI - [Psychosis as the initial manifestation of systemic lupus erythematosus: the role of lupus band test and anti-ribosomal antibodies]. AB - INTRODUCTION: Diffuse brain disease in systemic lupus erythematous (SLE) can be difficult to assess because of the sparse biological expression of the disorder, resulting in usually normal neuroimaging and laboratory findings. For this reason, it is likely that patients without a previous SLE diagnosis that presented initially with a pure psychiatric disorder, as psychosis or depression, can remain undiagnosed, and, in fact, they are exceptionally reported. As a biological marker, antiribosomal P protein antibodies have been closely related to SLE psychosis although their pathogenic role and specificity is under discussion. CLINICAL CASE: A young woman, without relevant medical history, presented with an acute psychotic catatonic picture, suspected organic in nature due to the existence of persistent mild CSF pleocytosis, while neuroimaging and laboratory studies were negative, including antinuclear and antiribosomal antibodies. Finally, a SLE diagnosis could be established because of a strong lupic band phenomenon in skin biopsy. CONCLUSIONS: It is important to maintain a high suspicion of SLE in acute psychotic patients, especially if atypical features are present. As the biological expression of the disorder in such cases may be elusive, we propose the study of lupus band in skin biopsy for these patients, regardless the absence of antiribosomal antibodies. PMID- 10363323 TI - [Clinical and prognostic heterogeneity in Aicardi's syndrome: a description of two cases]. AB - INTRODUCTION: Aicardi's syndrome is characterized by infantile spasms, agenesis of the corpus callosum and ocular lesions. Clinically it presents as severe mental retardation, severe limitation of motor development and of language, with a prognosis of survival for only a few months or years. We present two new cases of this uncommon syndrome and describe the heterogeneity of its clinical and prognostic severity. CLINICAL CASES: Case 1. A ten-month old patient had flexion spasms of the limbs at the age of 4 months, bilateral corioretinal lesions and generalized hypoplasia of the corpus callosum. During the clinical course of the disorder, the epileptic crises were controlled, there was mental retardation, the head was held steady and the baby could sit. Case 2. A nine year old patient had had flexion spasms when aged 2 months, had bilateral retinal lesions and generalized hypoplasia of the corpus callosum. During his clinical course the epileptic crises were controlled, there was severe mental retardation, the patient could pay attention and collaborate, articulate single words, walk on his own and manipulate objects. CONCLUSION: Aicardi's syndrome should be considered to be a syndrome in which the clinical findings and prognosis are heterogeneous, as seen from new cases with less clinical and functional limitation than the patients first described. PMID- 10363324 TI - [The pathological and the compensatory in cerebral ischemia: molecular aspects]. AB - INTRODUCTION: An integral approach of the whole mechanisms involved during ischemic brain damage let us to realize that some phenomena, usually considered as 'pathological' make an integrate and harmonious response directed towards the preservation of neuronal survival and functional integrity of the whole system. DEVELOPMENT: It has been proposed that cellular lysis in a core of lesion may be protective for the surrounding tissue, as well as the development of vasoconstriction and cellular edema, in the face of the loss of intracellular calcium homeostasis. Some recent findings tend to support this hypothesis. Adaptative response to lower oxygen and glucose availability is linked with the inhibition of neuronal activity, and therefore, with their involved functions. Formation of prostanoids and hydrogen peroxide, and release of interleukin and neurotrophic factors are related to the expression of immediate early genes, the enhancement of cellular response during ischemic conditions and also related to the induction of mechanisms of repair and regeneration, where astroglial cells play a key role. CONCLUSIONS: Pharmacological inhibition, hypothermia and hyperbaric oxygenation had been employed with a relative success in the clinical management of ischemic injured patients. That interventions are in agreement with some of these endogenous compensatory mechanisms, which become the brain less vulnerable that it is generally considered. The effectiveness of therapeutics in cerebrovascular disease must be increased if this compensatory response is adequately directed by physicians. PMID- 10363325 TI - [Neuropsychology and cognitive deterioration in epilepsy]. AB - OBJECTIVE: To carry out a critical evaluation of the international literature on the neuropsychology of epilepsy. The variables which lead to cognitive deterioration in epilepsy and the effect on cognition of different treatments available (pharmacological and surgical). DEVELOPMENT: We evaluate the influence of different neurological variables on the higher functions (aetiology, age of onset, type of crises, duration of illness, frequency and anti-epileptic drugs). We also describe the cognitive functions most affected in epilepsy (memory, attention, executive function, language). Studies of the surgery of epilepsy show that this may lead to both beneficial and undesirable effects on cognition. CONCLUSIONS: The variables which in general most affect higher functions are the duration of the illness and the frequency and types of crises. The commonest neuropsychological effects are those of memory deficit. These studies show that neuropsychological studies should be done from the time of onset of the disorder. Finally, from the surgical point of view, young patients with considerable crisis reduction or free of crises, with moderate preoperative deterioration of memory have the greatest possibility of post-operative improvement in cognition. Older persons with intact cognitive function and major surgical resection have more possibilities of post-surgical deterioration. PMID- 10363326 TI - [Motor complications of treatment with levodopa in Parkinson's disease]. AB - INTRODUCTION: Motor complications of treatment with levodopa affect more than 50% of patients after several years' treatment. This has a marked effect on patients with Parkinson's disease since these side effects are undesirable, affect the natural course of the disorder and complicate treatment. DEVELOPMENT: In this review we describe the main epidemiological, physiopathological and therapeutic aspects of the motor complications of levodopa. According to most epidemiological studies, starting treatment at an early age and more severe degree of the disorder are the main risk factors. The physiopathology of motor complications is varied. It has been thought to be due to loss of the buffer effect of the brain on plasma levels of levodopa, alterations in the functional state of the postsynaptic receptors and chronic intermittent use of levodopa. Management includes division and/or reduction of the dose of levodopa, use of slow-release preparations and association of dopaminergic agonists with COMT inhibitors which are the latest drugs to be tried. Various surgical possibilities in specialist centres may also be considered for serious cases of motor complications which do not respond to medical treatment. CONCLUSIONS: Recent advances in understanding the physiopathology and in the medical and surgical treatment permit improved control of the motor complications of chronic treatment with levodopa. PMID- 10363327 TI - [The penumbra area]. AB - INTRODUCTION: Habitually, when one speaks about penumbra area it refers to an ischemic region with the risk of permanent affection but potentially recoverable, that extend during a period of 4 to 6 hours. Nevertheless, with the reperfusion we cannot always get a neurofunctional recovery, or hinder the extension of the infarct. In this work, the author checked the mechanisms that participate in the lesion of penumbra area, as far as extension, duration as well as their relation with the therapeutic windows. DEVELOPMENT: Penumbra is a brain tissue at risk of infarct but is potentially recoverable and receives a variable level of cerebral blood flow (diminished, normal or augmented) which presents a functional alteration principally of its metabolism that is produced by various mechanisms like phenomenon of no reflow, reperfusion injury, hemodynamics disorders, spreading depolarization, delayed neural death, deafferentation (diaschisis), postischemic exofocal neural death, slowly progressive neural damage, among other alteration different a simple lesion by energy failure, these disorders may act during several months. CONCLUSION: Three therapeutics windows could be defined: one for the reperfusion (between 6 and 8 hours), another for the survival of neurons that are within the penumbra area (between 24 hour and 17 days) and a window for the neurofunctional recovery that extends itself to at least three months after a stroke. PMID- 10363328 TI - [Entacapone: is it useful as complimentary treatment with levodopa?]. AB - Entacapone (Comtan) is a potent, selective inhibitor of peripheral catechol-O methyltransferase (COMT) with therapeutic potential as an adjuvant to levodopa therapy in patients with Parkinson's disease. Entacapone decreases peripheral conversion of levodopa to 3-O-methyldopa increasing central extracellular levodopa and consequently striatal dopamine concentrations. At doses of 200 mg 2 to 10 times daily coadministered with levodopa/carbidopa or levodopa/benserazide entacapone may increase the duration of clinical response both after the first single dose and after repeated dosing in patients with end-of-dose fluctuations. At this dosage, it has a time to peak-plasma concentration of 1.2 hours and an elimination half life of 3.4 hours. In two multicentric, long-term (approximately 6 month), randomized and placebo-controlled studies, the duration of 'on' time was increased and the duration of 'off time' was decreased in patients who received adjunctive entacapone therapy. Moreover, patients randomized to entacapone reduced their levodopa requirements. In these and other phase III studies, entacapone was generally well tolerated, with few reported adverse events, mainly dyskinesias and gastrointestinal disorders. The dyskinesias were generally well controlled by decreasing the mean daily levodopa dose. Entacapone appears as a clinically significant and beneficial adjunct to levodopa therapy in Parkinson's disease patients with end-of-dose fluctuations. PMID- 10363329 TI - Exact and asymptotic tests for homogeneity in several 2 x 2 tables. AB - This paper presents the results of a Monte Carlo study comparing the performance, in terms of size and power, of six exact and six asymptotic tests for the homogeneity of odds ratios in several 2 x 2 contingency tables. With a small sample size or sparse data structure, the exact tests performed better than the asymptotic tests because they maintained the nominal size and, in some situations, had slightly higher power. Among the exact tests, we recommend the Zelen, Pearson chi-square and scores tests. Among the asymptotic tests, the Breslow-Day and Pearson chi-square tests were slightly better in some situations than the unconditional and conditional score tests. However, both exact and asymptotic tests had low power for small strata sizes, even with moderate to large heterogeneity of odds ratios. Corroborating previous findings, the asymptotic unconditional likelihood ratio test was too liberal in terms of size. PMID- 10363330 TI - Assessing heterogeneity and correlation of paired failure times with the bivariate frailty model. AB - We consider bivariate survival times for heterogeneous populations, where heterogeneity induces deviations in an individual's risk of an event as well as associations between survival times. The heterogeneity is characterized by a bivariate frailty model. We measure the heterogeneity effects through deviations associated with hazard functions and an association function defined through the conditional hazard functions: the cross-ratio function proposed by Oakes. We show how the deviation and association measures are determined by the frailty distribution. A Gibbs sampling method is developed for Bayesian inferences on regression coefficients, frailty parameters and the heterogeneity measures. The method is applied to a mental health care data set. PMID- 10363331 TI - A hierarchical Bayesian approach to age-specific back-calculation of cancer incidence rates. AB - We propose a Bayesian hierarchical model to estimate age-specific cancer incidence per year from age-specific cancer mortality. The model is based upon the empirical Bayesian approach of Liao and Brookmeyer (1995) and extends that model by consideration of the dependence on age. The incident cases per year are considered as observations from a discrete-time stochastic process following an autoregressive structure within a Poisson regression model. The model assumes that the survival probability among those with cancer is known. We have investigated the sensitivity of the model to the choice of this distribution and have found that this is the most sensitive part of the model. By comparison the predictions of the model are relatively robust to changes in other key areas, such as the number of years an incident case contributes before death, assumptions about parameter equality for identification and the initial prior distributions. The proposed methodology has been investigated using lung cancer mortality data from Scotland. Parameter estimates were obtained through Markov chain Monte Carlo methods, implemented using BUGS. PMID- 10363332 TI - Sample allocation for overlapping domains in a physician survey with a limited population. AB - We consider an analytic survey in which each survey unit can respond with respect to one or more domains to which it belongs. In this situation, the optimal sample allocation is constrained by the number of available units in each stratum defined by a set of domains. We present optimal sample allocation rules for this situation, and other rules that apply when there are additional constraints on sample sizes or variances by domain. We apply these rules to our motivating example, the design of a survey of physicians on their experiences with health plans, in which each physician can only be asked about her experience with one plan regardless of her number of affiliations. PMID- 10363333 TI - A comparison of mixed effects logistic regression models for binary response data with two nested levels of clustering. AB - We compare mixed effects logistic regression models for binary response data with two nested levels of clustering. The comparison of these models occurs in the context of developmental toxicity data sets, for which multiple types of outcomes (first level) are measured on each rat pup (second level) nested within a litter (third level). Because the nested nature of such data is occasionally accommodated by ignoring one level of clustering, we consider three models: (i) a three-level model adjusting for clustering due to both pup and litter (M1); (ii) a two-level model adjusting for just pup (M2); and (iii) another two-level model adjusting for just litter (M3). The three types of effects of interest are: (i) differences among malformation types (first-level effects); (ii) differences among groups of pups (for example, sex of pup, second-level effects); and (iii) differences among groups of litters (for example, dose, third-level effects). Simulations and data analyses suggest that the M3 model leads to more bias than the M1 or M2 models for all three types of effects. In terms of coverage of confidence intervals for fixed effects log odds ratio parameters, the M1 model achieves nominal coverage, whereas the M2 model reduces coverage for the third level effects and the M3 model obtains poor coverage for both first- and second level effects. These reductions in coverage for certain model-parameter combinations worsen as baseline risk increases. The data analyses support these simulation-based conclusions to some extent. PMID- 10363335 TI - A visualization of cross-over data using linear functions. AB - Previous work has shown that cross-over trial data can be analysed using within subject linear functions. Scores that result from linear functions are graphed in quantile comparison plots in order to visualize the differences between factor levels. An example suggests how this visualization can be used to identify outliers or to provide a more specific interpretation of results. Additional examples indicate how this approach can be used to track carry-over differences or interaction effects. This article is a US Government work and is in the public domain in the United States. PMID- 10363334 TI - Planning and analysis of repeated measures at key time-points in clinical trials sponsored by pharmaceutical companies. AB - In this paper we explore the possible reasons why medical papers reporting clinical trials sponsored by the pharmaceutical industry often analyse repeated measures data at certain key time-points instead of employing sophisticated models of repeated measures proposed by many statisticians. A survey indicated that the priority reason in the industry for having repeated measures in clinical trials is to monitor the trial and to utilize the early results for strategic decision making. We discuss what the common statistical methods do and do not offer for analysis of repeated measures in such clinical trials. We advocate the need to improve the understanding of the medical interest in conducting longitudinal trials in the industry, and to plan and analyse the repeated measures accordingly. We address the medical interest by formulating the problem and give illustrative examples for both phases II and III trials. PMID- 10363336 TI - Multivariate non-parametric methods for Mann-Whitney statistics to analyse cross over studies with two treatment sequences. AB - A non-parametric strategy for the analysis of ordinal data from cross-over studies with two treatment sequences and d(> or = 2) periods is examined through Mann-Whitney rank measures of association. For each period, these statistics estimate the probability of larger response for a randomly selected patient in one group relative to a randomly selected patient in the other group. Such estimates are as well formed for comparisons between groups for u pairs of periods with the same treatment. Methods for U-statistics are used to produce a consistent estimate of the covariance matrix for the (d + u) Mann-Whitney estimates. The effects of periods and treatments on the respective Mann-Whitney estimates are evaluated through linear (or log-linear) models. For estimation of the parameters in these models, a modified weighted least squares method is applied through a (2d - 1) < or = (d + u) dimensional basis which effectively addresses potentially near singularities in the estimated covariance matrix of the Mann-Whitney estimates. The proposed methods are applicable to response variables with an interval or an ordered categorical scale. Their scope additionally has capabilities for controlling strata in the design of a cross over study or concomitant variables for which covariance adjustment is of interest for reduction of variance. Applications of the methods are illustrated through three cross-over studies with different specifications for the two sequences of two treatments during two to four periods. PMID- 10363337 TI - Accrual strategies for phase I trials with delayed patient outcome. AB - Phase I dose-finding trials typically are conducted using adaptive rules that select dose levels for successive patient cohorts based on the outcomes of patients treated previously in the trial. When patient outcome cannot be observed immediately after treatment, the problem arises of how to deal with new patients while waiting to observe the current patient cohort's outcomes. We consider two alternative approaches to this problem in the context of a phase I trial conducted using the continual reassessment method. With the first approach, a patient requiring treatment before the next cohort opens is treated off protocol with standard therapy, and otherwise waits until the next cohort opens. The second approach treats each patient immediately upon arrival at the dose recommended based on currently available data. We compare these two approaches by simulation under varying dose--toxicity curves, accrual rates, cohort sizes and early stopping rules. We evaluate patient waiting time, trial duration, number of patients treated off protocol and the probabilities of toxicity and of selecting the correct dose. We also study three strategies for assigning patients to trials when two or more phase I trials may be ongoing simultaneously. Based on our results, we provide practical guidelines for deciding among these approaches and strategies in a given clinical setting. PMID- 10363338 TI - Comparing two prevalence rates in a two-phase design study. AB - An epidemiological study often uses a two-phase design to estimate the prevalence rate of a mental disease. In a two-phase design study, the first phase assesses a large sample with an inexpensive screening test, and then the second phase selects a subsample for a more expensive diagnostic evaluation. Furthermore, disease status may not be ascertained for all subjects who are selected for disease verification because some subjects are unable to be clinically assessed, while others may refuse. Since not all screened subjects are selected for diagnostic assessments, there is potential for verification bias. In this paper, we propose the maximum likelihood (ML) and bootstrap methods to correct for verification bias for estimating and comparing the prevalence rates under the missing-at-random (MAR) assumption for the verification mechanism. We also propose a method to test this MAR assumption. Finally, we apply our methods to a large-scale prevalence study of dementia disorders. PMID- 10363339 TI - A strategic view of randomized trial design in low-incidence paediatric cancer. AB - We use a statistical model to examine the relationship between alpha level, sample size, trial duration, patient accrual rate and therapeutic innovation rate on the increase in treatment efficacy achieved after a series of two-treatment randomized phase III trials. In a setting where the trials include most of the patients in the target population for inference, as in some paediatric cancers, we show that the traditional criteria by which one determines trial size are difficult to justify and apply. In particular, using as a measure of evidence type I error levels larger than the typical 5 per cent for judging treatment differences, and performing smaller trials than one would usually consider feasible, yields on average, over a 25-year research course, larger gains in cure rate. Judicious choice of type I error rate and trial size keeps the chance of worsening treatment efficacy at a low level, even while increasing the chance of making large improvements in cure rate. We propose that a more appropriate view of trial design in low-incidence cancer settings is in the overall context of the research setting and long-term goals rather than in the narrow context of the current single trial. From this viewpoint, insistence on large trials and stringent evidence for accepting new treatments can be counter-productive, in that likely gains in efficacy of treatment will be smaller over the long term. PMID- 10363340 TI - Exact test size and power of a Gaussian error linear model for an internal pilot study. AB - Wittes and Brittain recommended using an 'internal pilot study' to adjust sample size. The approach involves five steps in testing a general linear hypothesis for a general linear univariate model, with Gaussian errors. First, specify the design, hypothesis, desired test size, power, a smallest 'clinically meaningful' effect, and a speculated error variance. Second, conduct a power analysis to choose provisionally a planned sample size. Third, collect a specified proportion of the planned sample as the internal pilot sample, and estimate the variance (but do not test the hypothesis). Fourth, update the power analysis with the variance estimate to adjust the total sample size. Fifth, finish the study and test the hypothesis with all data. We describe methods for computing exact test size and power under this scenario. Our analytic results agree with simulations of Wittes and Brittain. Furthermore, our exact results apply to any general linear univariate model with fixed predictors, which is much more general than the two-sample t-test considered by Wittes and Brittain. In addition, our results allow for examination of the impact on test size of internal pilot studies for more complicated designs in the framework of the general linear model. We examine the impact of (i) small samples, (ii) allowing the planned sample size to decrease, (iii) the choice of internal pilot sample size, and (iv) the maximum allowable size of the second sample. All affect test size, power and expected total sample size. We present a number of examples including one that uses an internal pilot study in a three-group analysis of variance. PMID- 10363341 TI - Estimation and comparison of rates of change in longitudinal studies with informative drop-outs. AB - Many cohort studies and clinical trials have designs which involve repeated measurements of disease markers. One problem in such longitudinal studies, when the primary interest is to estimate and to compare the evolution of a disease marker, is that planned data are not collected because of missing data due to missing visits and/or withdrawal or attrition (for example, death). Several methods to analyse such data are available, provided that the data are missing at random. However, serious biases can occur when missingness is informative. In such cases, one needs to apply methods that simultaneously model the observed data and the missingness process. In this paper we consider the problem of estimation of the rate of change of a disease marker in longitudinal studies, in which some subjects drop out prematurely (informatively) due to attrition, while others experience a non-informative drop-out process (end of study, withdrawal). We propose a method which combines a linear random effects model for the underlying pattern of the marker with a log-normal survival model for the informative drop-out process. Joint estimates are obtained through the restricted iterative generalized least squares method which are equivalent to restricted maximum likelihood estimates. A nested EM algorithm is applied to deal with censored survival data. The advantages of this method are: it provides a unified approach to estimate all the model parameters; it can effectively deal with irregular data (that is, measured at irregular time points), a complicated covariance structure and a complex underlying profile of the response variable; it does not entail such complex computation as would be required to maximize the joint likelihood. The method is illustrated by modelling CD4 count data in a clinical trial in patients with advanced HIV infection while its performance is tested by simulation studies. PMID- 10363342 TI - Interval-censored survival data with informative examination times: parametric models and approximate inference. AB - We develop parametric methods for analysing interval-censored data when examination and survival times are not independent. The hazard function is modelled by introducing individual frailties related to the frequency of examinations. Model parameters may be obtained by direct maximization of the marginal log-likelihood. We develop a simpler approximate method in which the frailties are estimated by empirical Bayes. The two approaches are equivalent asymptotically as the number of examinations on each individual increases. Simulations suggest that the approximate method is adequate for estimating regression parameters even when the number of examinations on each individual is small. The methods are used to estimate age and period effects on HIV incidence in a cohort of repeat attenders at genito-urinary clinics in London, U.K. PMID- 10363343 TI - A discriminant analysis extension to mixed models. AB - Discriminant analysis is commonly used to classify an observation into one of two (or more) populations on the basis of correlated measurements. Classical discriminant analysis approaches require complete data for all observations. Our extension enables the use of all available longitudinal data, regardless of completeness. Traditionally a linear discriminant function assumes a common unstructured covariance matrix for both populations, which may be taken from a multivariate model. Here, we can model the correlated measurements and use a structured covariance in the discriminant function. We illustrate cases in which the estimated covariance structure is either compound symmetric, heterogeneous compound symmetric or heterogeneous autoregressive. Thus a structured covariance is incorporated into the discrimination process in contrast to standard discriminant analysis methodology. Simulations are performed to obtain a true measure of the effect of structure on the error rate. In addition, the usual multivariate expected value structure is altered. The impact on the discrimination process is contrasted when using the multivariate and random effects covariance structures and expected values. The random-effects covariance structure leads to an improvement in the error rate in small samples. To illustrate the procedure we consider repeated measurements data from a clinical trial comparing two active treatments; the goal is to determine if the treatment could be unblinded based on repeated anxiety score measurements. PMID- 10363344 TI - Joint effects on cancer risk of age at childbirth, time since birth and attained age: circumventing the problem of collinearity. AB - In previous studies of female cancer risk, we introduced a new method for circumventing the problem of collinearity in age-adjusted analysis of the joint effects of age at birth and time since birth. The basic idea was to estimate the pure age effect considering nulliparous women, assuming that the age effect is common to all women. However, risk estimates for attained age obtained in this manner may suffer from bias, in particular in small data sets, which may in turn influence risk estimates for reproductive factors among parous women. Certain factors possibly affecting cancer risk among nulliparous women only, for instance biological infertility, might also introduce bias. The purpose of this paper is to investigate the accuracy of risk estimates obtained by the joint approach, and to reveal the extent of bias in traditional separate age-adjusted analyses of age at birth or time since birth among parous women. Results are based on analyses of simulated data sets reflecting reproductive and demographic characteristics of a cohort of 1.1 million Norwegian women. Incidence rate ratios are calculated in Poisson regression analyses of person-years at risk. Our simulations show that the joint analysis in general yields unbiased risk estimates, but the number of cases must be rather high to achieve reliable results. Risk estimates from separate analyses can be seriously biased, although the amount of bias depends on the strength and direction of associations with cancer risk. With a total of 5500 cancer cases, the estimators for age at last birth and time since last birth were 13-78 per cent and 5-66 per cent more efficient in the joint than in the separate analysis, respectively. Significance levels were close to the nominal 5 per cent in the joint analysis, but about twice as high in the separate analysis. Adding an effect of biological infertility on cancer risk among nulliparous women, without taking it into account in the analyses, did not seriously affect risk estimates in the joint model. PMID- 10363345 TI - Process control procedures to augment quality control of leukocyte-reduced red cell blood products. AB - Quality control of leukocyte-reduced packed red cell units (LRprc) produced in blood facilities must conform to regulatory criteria, which state that units may not contain more than 1 x 10(6) to 5 x 10(6) white blood cells (WBC) per unit. The post-filtration WBC content of a total of n = 386 LRprc units was counted with a Nageotte chamber to model the probability that a unit would not meet the regulatory criteria. The distribution of the residual leukocyte counts is close to a negative binomial distribution (NBD) and is independent of the packed red cell volume filtered. The observed probability that a unit of blood has a residual WBC greater than 5 x 10(6) is 2.6 +/- 2.6 x 10(-3). A power analysis of the two-sample Kolmogorov-Smirnov (KS) test in this application shows that a sample size of 20 is sufficient for determining that the process is in control when an out of control process has a k NBD parameter greater than or equal to that of the in control process. The three out of control processes observed to date appear to have this property. A sample of size 80 may be necessary for confirming that process validation data sets conform to the larger 'reference' database (n = 386) for processes that are out of control in such a way that their k NBD parameter is less than the k parameter of the in control process. PMID- 10363346 TI - Attention for action: coordinating bimanual reach-to-grasp movements. AB - Theories of attention have frequently pointed to the finding that there is a significant performance decrement ('cost') to responding to two different objects concurrently. However, much of the research aimed at investigating how attention is 'divided' in such circumstances has adopted response time (RT) as the measure of interest. In this paper we investigate how attention is 'divided' during the execution of concurrent motor responses, by studying bimanual reach-to-grasp movements directed towards two separate target objects. Furthermore, a key aspect of our study is that each hand is required to perform either the same action (congruent reaches) or a different action (incongruent reaches). Thus in Expt 1 we manipulated the movement amplitude of each hand, while in Expt 2 we manipulated object size. The results of this study suggest that while there is an overall cost associated with carrying out two movements simultaneously, kinematic measures are unaffected by whether the actions required of each hand are the same (congruent) or different (incongruent). The problem of executing incongruent bimanual movements appears to be solved by synchronizing each limb to a common movement duration, while movement velocity and grip aperture are independently scaled. These findings are discussed in relation to theories developed to explain the coordination of the reach-and-grasp phases of unimanual prehension, and in the context of recent theories of attention for action. PMID- 10363347 TI - The coordination of two-effector actions: spoon-feeding and intermanual prehension. AB - The aim of this study was to assess whether kinematic characteristics of unimanual (one-effector) prehension movements can also be found in tasks where the transport and grasp components are distributed across separate effectors. Expt 1 compared prehension with spoon-feeding and showed that a number of kinematic landmarks, such as time to peak aperture, were similar. An unexpected difference between the tasks was that the onsets of the transport and grasp components in the two-effector task were not always synchronized. To control for a number of differences between the tasks used in Expt 1, such as the role of haptic and visual information, Expt 2 compared unimanual with intermanual prehension, which involved passing an object from one hand to the other. The results were consistent with those found in Expt 1. In particular, time to peak aperture was similar over the different tasks, despite a lack of consistent coupling between the components at movement onset. We conclude that the one- and two-effector tasks pose largely the same task constraints and are coordinated in a similar way. Intermanual prehension tasks thus provide a useful experimental tool for manipulations that are difficult or impossible to perform with unimanual reaching and grasping. PMID- 10363348 TI - On the perceived time of voluntary actions. AB - When do we think we move when we make a voluntary action? Previous studies have pointed to an anticipatory awareness of action (i.e. we think we move before we actually do), but have not investigated the content or locus of motor awareness. In the experiment reported here the authors localize the time of awareness of the first movement in a sequence within the context of a specific information processing model. The results suggest that our awareness of our own actions is associated with some pre-motor event after the initial intention and preparation of action, but before the assembly and dispatch of the actual motor command to the muscles. PMID- 10363349 TI - Parent-child mediated learning interactions as determinants of cognitive modifiability: recent research and future directions. AB - The main objectives of this article are to describe the effects of mediated learning experience (MLE) strategies in mother-child interactions on the child's cognitive modifiability, the effects of distal factors (e.g., socioeconomic status, mother's intelligence, child's personality) on MLE interactions, and the effects of situational variables on MLE processes. Methodological aspects of measurement of MLE interactions and of cognitive modifiability, using a dynamic assessment approach, are discussed. Studies with infants showed that the quality of mother-infant MLE interactions predict later cognitive functioning and that MLE patterns and children's cognitive performance change as a result of intervention programs. Studies with preschool and school-aged children showed that MLE interactions predict cognitive modifiability and that distal factors predict MLE interactions but not the child's cognitive modifiability. The child's cognitive modifiability was predicted by MLE interactions in a structured but not in a free-play situation. Mediation for transcendence (e.g., teaching rules and generalizations) appeared to be the strongest predictor of children's cognitive modifiability. Discussion of future research includes the consideration of a holistic transactional approach, which refers to MLE processes, personality, and motivational-affective factors, the cultural context of mediation, perception of the whole family as a mediational unit, and the "mediational normative scripts." PMID- 10363350 TI - The negative impact of the cognitive movement on the continued growth of the behavior therapy movement: a historical perspective. AB - In recent years, a growing number of behavior therapists have expressed concern over the current state of the behavioral therapy movement. Some of the major problems raised center on current overload and fractionization, the lack of a coherent overall picture, the loss of identity, and the influx of cognitivism. In an attempt to enhance understanding of the factors responsible for the current crises in the behavior therapy field, the author provides a historical overview of the behavioral movement from its original conception to its current state. An argument is made that the solution to the afore-mentioned problems resides in the readoption of the underlying philosophy of science that originally gave birth and purpose to the field. PMID- 10363351 TI - Consequences of adolescent marijuana use: incompatibility with the assumption of adult roles. AB - This longitudinal study is an examination of the relationship between marijuana use and the assumption of adult roles, as well as the relationship between assuming adult roles and the likelihood of later marijuana use. Data were collected at 5 points in time from childhood through early adulthood (late 20s) by means of a structured questionnaire. Participants' marijuana use and the assumption of adult roles, including employment, marriage, parenthood, and living arrangements, were measured, and the data were analyzed with logistic regression analyses. A history of marijuana use was associated with an increased risk of adopting more unconventional adult roles, such as postponement of marriage, having a child out of wedlock, and unemployment. These results suggest that frequent prior marijuana use may adversely affect one's ability to successfully assume conventional adult roles. Furthermore, controlling for earlier marijuana use, marriage during early adulthood significantly decreased the risk of later marijuana use. PMID- 10363352 TI - Apoptosis and differentiation in the crypt-villus unit of the rat small intestine. AB - This investigation utilized immunocytochemical and fluorescent protocols to analyze the roles of cellular proliferation and apoptosis in the regulation of differentiation and senescence of the rat small intestinal mucosa. Specifically, the study localized apoptotic zones of the villus through the use of enzymatic tags; established the transition point between cell growth and differentiation, i.e. the point of no return where crypt cells differentiate into absorptive cells with barrier functions; and the role that plasmalemmal, cytoskeletal, junctional and extracellular matrix (ECM) elements may play in the regulation of differentiation and migration of epithelial cells from crypt to villus. Apoptosis was relegated to the villus tip forming a prominent 'apoptotic cuff' of cells. Close scrutiny of these cuffs reveals the presence of apoptotic cells adjacent to non-apoptotic (healthy) cells. Mid-villus epithelial cells were non-apoptotic and all cells in the crypt-villus unit expressed Bcl-2 activity. Intestinal lactase expression was prominent in post-mitotic cells along the villus, while cells in the crypt and base were negative for lactase activity. In contrast, all the cells of the crypt-villus unit were intensely reactive for F-actin. Close scrutiny of isolated cells and frozen sections indicates specific localization of actin in the microvillus region, apical cytoplasm, basolateral and lateral plasmalemma which was in close proximity to fibronectin in the basement lamina. Occludin positive junctional networks were prominent at villus tips, where senescent and apoptotic cells were also most prominent, suggesting that tight junctional integrity was essential to barrier, digestive and absorptive functions in all regions of the mucosa. PMID- 10363353 TI - Control of neuron outgrowth by NMDA receptors. AB - The role of N-methyl-D-aspartic acid receptors (NMDARs) of glutamate on neuritogenesis was studied in cultured neurons of chick embryo spinal cord using the NMDAR non-competitive antagonist dizocilpine maleate (MK-801). No cell population was fully prevented from neuritogenesis by MK-801. Different aspects of neuritogenesis were quantitatively evaluated. Neurite initiation, elongation and branching were depressed by MK-801. Inhibition was dose-dependent and reversible. A loss of responsiveness of neuritogenesis to MK-801 was found during the second day of treatment at a concentration of 10 microM, but not at higher concentrations. Our findings support the idea that Ca2+ influx through NMDAR associated channels is one of the possible triggers of a cascade resulting in neuritogenesis. The effects of NMDAR blocking on neuritogenesis occurred before synaptogenesis, suggesting a role of excitatory aminoacids in neuron morphological differentiation at early stages of development. Scanning electron microscopy confirmed a reduction in neurite tree complexity in MK-801 treated cells and showed a production of filopodium-like processes in some of these cells. PMID- 10363355 TI - Pancreas ultrastructural alterations in mice inoculated with Tityus discrepans (Buthidae) venom. AB - The symptoms of scorpionic envenomation in mice appear almost immediately after intraperitoneal injection and are manifested by great agitation, hair bristling, accelerated respiration, salivation and lacrimation, vomits and diarrhoea. In this work we intend to correlate those clinical manifestations appearing in response to the toxic aggression by Tityus discrepans venom, to the cellular or subcellular alterations produced in the mouse pancreas, probably similar to those damages found in envenomed humans. To evaluate pancreas subcellular response to Tityus discrepans venom, male C57/Bl adult mice were randomised into two groups: envenomed were intraperitoneally injected (hypochondrial left region) at a dose of 5 mg/Kg weight and controls received saline solution. Samples after preparation were studied in a Hitachi-300 transmission electron microscope. The most relevant ultrastructural changes in pancreatic tissues were an increase in the nuclear heterochromatin, with a corresponding decrease of euchromatin. In the cytoplasm, rough endoplasmic reticulum exhibited zones of oedema, losing its organised aspect. The secretion granules presented smaller electron density and variability in dimensions. At higher magnification a nucleus with picnotic appearance, with indentation of its perinuclear cistern was observed. There was a mitochondrial degeneration, with destruction of the mitochondrial matrix and autophagic vacuoles in its interior. At 48 h the lesions became intensified, with an evident increase in the intercellular spaces. PMID- 10363354 TI - Molecular pathology of Luft disease and structure and function of mitochondria. AB - Luft disease, studied in detail by Luft et al. (1962) is characterized clinically by hypermetabolism and consequent abnormal transpiration. In their study, Luft and coworkers revealed that the hypermetabolism is caused by extensive uncoupling of mitochondrial respiration in skeletal muscle tissue. They also discovered that the muscle mitochondria had been structurally modified with the cristae assuming a zig-zag conformation. The mitochondrial enzymes functioned normally, the abnormality being confined to the extensive uncoupling of respiration. In an earlier study (Sjostrand et al., 1988) it was revealed that the zig-zag conformation is caused by removal of some tricarboxylic acid cycle enzymes from the cristae, exposing the interior of the cristae to the matrix fluid. Applying a coupling theory proposed by Sjostrand (1990, 1991), leakage of protons from the cristae should impair coupling of respiration and ATP synthesis. The structural damage is conceived of to be caused by a genetic defect preventing proper aggregation of the enzyme molecules in the cristae. Luft disease may therefore be the first known disease caused primarily by a structural disorder at the molecular level. The symptoms of the disease reveal the importance of the compact and ordered aggregation of the enzyme molecules in the cristae for coupling of respiration. PMID- 10363356 TI - Distribution of sodium and potassium channels as well as myelin associated glycoprotein (MAG) during the early stages of Wallerian degeneration. AB - The distribution of sodium and potassium channel proteins and the myelin associated glycoprotein (MAG) was studied by immunofluorescence during the early stages of Wallerian degeneration. Routine electron microscopy was also performed in order to investigate the success of the lesion in producing degeneration and also to evaluate the integrity of the axolemma and cytoskeleton. Sural nerves from Sprague-Dawley rats were submitted to surgical crush and analyzed after 30, 36 and 48 h. The preparations were observed by light microscopy and the amount of labeled and unlabeled sites was quantified using a computer linked microscope. The number of sodium and potassium labeled nodes was dramatically reduced 30 h after crushing. However, a small number of labeled nodes was still present even after 48 h. These remaining nodal channel proteins are probably responsible for the maintenance of the nerve's electrical activity during the first 4 days of Wallerian degeneration. Ultrastructural analyses of longitudinal sections revealed areas of intact axolemma in the presence of a partially or completely disrupted cytoskeleton. These results are in disagreement with the generally accepted view that axolemma and cytoskeleton disruption occur simultaneously in the peripheral nervous system. Our results also concern the mechanism underlying the disappearance of these channel proteins during the degeneration of the peripheral nerve fibres, since in this pathology the axons are affected before myelin and Schwann cells become affected. PMID- 10363357 TI - Regeneration of supraependymal nerve fibers in rat. AB - Factors intrinsic and extrinsic to the neurons regulate the regeneration of axons following axotomy. Elucidation of these mechanisms will provide fundamental information on the regulation of growth in mammalian neurons. To understand the role of environmental factors in nerve regeneration, this study focuses on the axotomy-induced regeneration of supraependymal nerve fibers that inhabit an environment different from other central nervous system neurons and the neurons of the peripheral nervous system. These fibers are ideally located to receive various neuroactive substances that are streaming in the cerebrospinal fluid. Transmission electron microscopic studies reveal the presence of three types of fibers on the ventricular surface. They are fibers with clear synaptic vesicles, those containing small dense core vesicles together with clear synaptic vesicles and fibers containing large dense core granules. To study the regenerative potential of supraependymal nerve fibers, they were axotomized by inflicting mechanical injury to the floor of the third ventricle. Following axotomy, the regenerative ability of the supraependymal nerve fibers was monitored between five and thirty days, by scanning electron microscopy. By the fifth day, there was a noticeable increase in the neuronal network that gradually increased throughout the experimental period. The regenerative ability of supraependymal neurons, as compared with other neurons in the central nervous system, could be due to the differences in their environment. These studies suggest that mammalian cerebral ventricles may be an adequate site for neural transplantation. PMID- 10363358 TI - Scanning electron microscopy of stomach and small intestine of rabbit during foetal and post natal life. AB - The surface pattern of the stomach and the small intestine of the rabbit was examined using SEM in stages ranging from 26 day of foetal to 24 days of post natal life. The stomach glands were evident in the late foetuses and became gradually deeper after birth. The villi of the small intestine appeared to have a gradual development during foetal and post-natal life, being short and thick with rounded tips during foetal life while becoming long, slender and finger-like with increasing age. PMID- 10363359 TI - Lectin binding sites in parotid acinar secretory granules of normal and isoproterenol treated rat. AB - Lectin staining patterns in secretory granules of rat parotid gland acinar cell of untreated and isoproterenol-injected animals were examined by electron microscopy. We used four lectin-gold complexes: Ulex europaeus agglutinin I (UEA I), Helix pomatia agglutinin (HPA), wheat germ agglutinin (WGA), Glycine max agglutinin (SBA). Specimens were low temperature embedded in the hydrophilic Lowicryl K4M resin. The normal acinar cells produced glycoconjugates which were positive for all of the lectins used and with a characteristic topographic distribution in relation to the morphological type of granule. The cells of isoproterenol-treated rat showed marked ultrastructural changes in the size and structure of granules; significant changes in lectin binding sites in the granules were also observed. PMID- 10363360 TI - Ultrastructural localization of anionic sites and lectin-binding sites in sarcoid human alveolar macrophages during interaction with T-lymphocytes. AB - Sarcoidosis alveolitis is caused by an unknown stimulus activating alveolar macrophages (AM) and T-lymphocytes. During antigen presentation, the complex HLA class II molecule/processed peptide, on the surface of sarcoid AM, induces the T lymphocyte to proliferate. Altered glycosylation patterns of cell surface glycoproteins such as class II molecules in inflammatory states, may enhance the antigen-presenting capability of AM. In order to know if anionic sites and lectin binding sites take part in the process of antigen presentation by alveolar macrophages, cells obtained from bronchoalveolar lavage of patients with pulmonary sarcoidosis were incubated with cationized ferritin (CF) and colloidal gold complexed lectins (BSL-I-A4; RCA-I; RCA-II; WGA) for 30 min at 4 degrees C. After incubation, the cells were fixed with 4% paraformaldehyde, 2% glutaraldehyde, postfixed, and Epon embedded. The CF particles were uniformly distributed over the entire cell surface of the lymphocyte, and formed clusters on the surface of the macrophage mainly at the adhesion region between the AM and the lymphocytes. We found enhanced binding of BSL-I-A4 by AM, while WGA and RCA were poorly taken up by these cells. Gold-BSL-I-A4 was distributed randomly on the plasma membrane of the AM, and clustered in the adhesion region with lymphocytes. These results suggest that anionic sites and alpha-D-N-acetyl galactosamine residues labeled with gold-BSL-I-A4 may be involved in the process of antigen presentation by sarcoid alveolar macrophages. PMID- 10363361 TI - Attenuation of Ca paradox injury in guinea pig heart by K+ channel blocker, d sotalol. AB - D-sotalol was shown to prevent Ca overload and intermyocyte uncoupling. The aim of this study was to investigate the effect of d-sotalol in Ca paradox conditions. Guinea pig hearts were perfused at 37 degrees C and constant pressure with oxygenated Tyrode solution. Ca paradox was induced by 10 min Ca free perfusion followed by 10 min Ca repletion. 10(-6) M d-sotalol was administered either during Ca depletion or during Ca repletion period. Electrical activity and ventricular contraction were simultaneously recorded and subcellular alterations were analysed. The contraction terminated in 5 min of Ca free perfusion and electrical activity disappeared within 5 min of Ca repletion. Nonuniform injury of myocardial tissue was observed. The majority of cardiomyocytes were irreversibly injured and profound dissociation of intercellular junctions was detected. Administration of d-sotalol during Ca free period preserved electrical activity and restored ventricular contraction accompanied by apparent protection of the ultrastructure, including intercellular connections. Uniform patterns of sarcomeres reflected synchronous contraction and protection of junctional couplings. In conclusion, d-sotalol attenuates Ca paradox injury. It seems that the protective effect of d-sotalol is most likely related to inhibition of potassium efflux antagonizing Na loading during Ca depletion period, as well as to attenuation of excess of [Ca2+]i via acceleration of sarcoplasmic Ca exchange during Ca repletion. PMID- 10363362 TI - Ultrastructure of the blood-brain barrier in the rabbit. AB - The morphology of the normal blood-brain barrier in the rabbit by thin section and freeze-fracture electron microscopy is reported. Exogenous tracer horseradish peroxidase was injected to visualize the integrity of the blood-brain barrier in New Zealand White rabbits. Freeze-fracture was used to determine the intramembrane architecture of the tight junctions. Thin sections (60-100 nm) of brain capillaries from animals injected with horseradish peroxidase (HRP) possessed few pinocytotic vesicles in the cytoplasm. Junctional profiles between adjoining plasma membranes were present. Thin sections of capillaries containing electron dense HRP reaction product (HRP-RP) in the lumen revealed focal fusions of apposing plasma membranes that occluded reaction product from entering the junctional clefts. Some cytoplasmic vesicles were filled with HRP-RP; however, basal laminae and brain interstitium were free of HRP-RP in all vessel profiles examined. Freeze-fracture electron microscopy revealed tight junctions as an elaborate network of interconnecting strands of intramembrane particles appearing as ridges on the EF face and corresponding grooves on the PF face on platinum replicas. Results of this study demonstrate the architecture of rabbit brain microvessel endothelial junctions (blood-brain barrier) and provide evidence that the tight junctions prevent HRP extravasation. It is concluded that rabbit brain endothelial tight junctions (zonulae occludentes), as in other mammals, form the anatomical basis of the blood-brain barrier. Consequently, the rabbit brain microvasculature can be a useful model for establishing stereotactic radiosurgical procedures to treat brain astrocytomas (tumours). PMID- 10363363 TI - Ultrastructural aspects of the retractor ocular bulbi muscle in the opossum (Didelphis albiventris). AB - In the present study the different types of muscle fibers of the retractor ocular bulbi muscle of the South American opossum were classified according to their ultrastructural characteristics. The tridimensional characteristics of the neuromuscular junctions present in this muscle were also demonstrated by scanning electron microscopy (SEM). Five adult opossums, three males and two females, were perfused with fixative solution through the left ventricle and their right retractor ocular bulbi muscle was prepared for the ultrastructural study of muscle fibers. The contralateral muscle was used for the study of neuromuscular junctions by SEM. Three types of fibers were detected, denoted 1r, 2r and 3r. Only simple neuromuscular junctions of the plate type were visualized by SEM. PMID- 10363364 TI - Primary cilium expression in cells from normal and pathological caprine skin. AB - Primary cilia were detected in keratinocytes and fibroblasts, from the skin of two healthy and five Saanen goats suffering from a severe papillomatosis of the udder, respectively. Single cilia were detected in very few normal and neoplastic keratinocytes; rare biciliated keratinocytes were also seen. A remarkable number of dermal fibroblasts from healthy goats showed single cilia. Similarly, primary cilia were found in fibroblasts from the tumour stroma in all five goats. These data seem to strengthen the statement that ciliation is a peculiar ultrastructural aspect of fibroblasts, which is of interest in the light of the emerging role of fibroblasts in many physiopathological processes. PMID- 10363365 TI - Rice ENOD40: isolation and expression analysis in rice and transgenic soybean root nodules. AB - The early nodulin ENOD40 has been proposed as playing a pivotal role in the organogenesis of legume root nodules. We have isolated the ENOD40 gene homologues ObENOD40 and OsENOD40 from the wild and cultivated rice genotypes Oryza brachyantha and Oryza sativa, respectively. Rice ENOD40s contain a sequence at the 5' end (region I) for encoding an oligopeptide that is highly conserved in all legume ENOD40s. Furthermore, at the 3' end (region II), the nucleotide sequence of rice ENOD40s exhibited a considerable homology to the corresponding region in legume ENOD40s. Among various organs of the rice plant, expression of OsENOD40 was detected only in stems. In situ hybridization studies revealed that, within the stem, transcription of OsENOD40 is confined to parenchyma cells surrounding the protoxylem during the early stages of development of lateral vascular bundles that conjoin an emerging leaf. Expression pattern of OsENOD40 promoter-GUS fusion in nodules developed on transgenic hairy roots of soybean was also found to be restricted to peripheral cells of nodule vascular bundles, thus evidencing that rice ENOD40 promoter activity is essentially the same as that of soybean ENOD40. Taken together, these results strongly suggest that OsENOD40 and legume ENOD40s share common, if not identical, functions in differentiation and/or function of vascular bundles. PMID- 10363366 TI - A chromosomal paracentric inversion associated with T-DNA integration in Arabidopsis. AB - T-DNA integration in the nuclear plant genome may lead to rearrangements of the plant target site. Here we present evidence for a chromosomal inversion of 26 cM bordered by two T-DNAs in direct orientation, which is linked to the mgoun2 mutation. The integration sites of the T-DNAs map at positions 80 and 106 of chromosome I and we show that each T-DNA is bordered by plant sequences from positions 80 and 106, respectively. Although the T-DNAs are physically distant, they are genetically closely linked. In addition, three markers located on the chromosome segment between the two T-DNA integration sites show no recombination with the mgo2 mutation. We show that the inversion cannot be a consequence of a recombination event between the two T-DNAs, but that the integration of the T DNAs and the inversion were two temporally linked events. T-DNA integration mechanisms that could have led to this inversion are discussed. PMID- 10363367 TI - Sucrose transport into developing seeds of Pisum sativum L. AB - The anatomy of developing pea seeds is characterized by transfer cells present in both coats and cotyledons at the maternal/filial interface. To determine the nature and cellular localization of sucrose transporters in pea seeds, a full length clone of a sucrose/H+ symporter (PsSUT1) was isolated from a cotyledon cDNA library. Northern blot analyses of different organs showed that PsSUT1 is expressed in non-seed tissues, including sucrose sinks and sources. Within developing seeds, transcripts of PsSUT1 and PsAHA1 genes were detected in all tissues, while transcripts of a sucrose binding protein (GmSBP) were confined to cotyledon epidermal transfer cells. Signal intensities of PsSUT1 and PsAHA1 transcripts and protein products were most pronounced in the thin-walled parenchyma cells of seed coats and epidermal transfer cells of cotyledons. For cotyledons, the highest transporter densities were localized to those portions of plasma membranes lining the wall ingrowth regions of epidermal transfer cells. Responses of [14C]sucrose influx to metabolic inhibitors indicated that proton coupled sucrose transport was operative in both seed coats and cotyledons. Cotyledon epidermal transfer cells were shown to support the highest sucrose flux. Maximal transport activity was found to account for the sucrose flux differences between seed tissues. Intercellular movement of the symplasmic tracer, 5-(6)-carboxyfluorescein (CF), demonstrated that symplasmic pathways interconnect the vascular tissues to thin-walled parenchyma transfer cells of seed coats and, for cotyledons, epidermal transfer cells to storage parenchyma cells. PMID- 10363368 TI - A minor form of starch branching enzyme in potato (Solanum tuberosum L.) tubers has a major effect on starch structure: cloning and characterisation of multiple forms of SBE A. AB - Full length cDNAs encoding a second starch branching enzyme (SBE A) isoform have been isolated from potato tubers. The predicted protein has a molecular mass of 101 kDa including a transit peptide of 48 amino acids. Multiple forms of the SBE A gene exist which differ mainly in the length of a polyglutamic acid repeat at the C-terminus of the protein. Expression of the mature protein in Escherichia coli demonstrates that the gene encodes an active SBE. Northern analysis demonstrates that SBE A mRNA is expressed at very low levels in tubers but is the predominant isoform in leaves. This expression pattern was confirmed by Western analysis using isoform specific polyclonal antibodies raised against E. coli expressed SBE A. SBE A protein is found predominantly in the soluble phase of tuber extracts, indicating a stromal location within the plastid. Transgenic potato plants expressing an antisense SBE A RNA were generated in which almost complete reductions in SBE A were observed. SBE activity in the leaves of these plants was severely reduced, but tuber activity was largely unaffected. Even so, the composition and structure of tuber starch from these plants was greatly altered. The proportion of linear chains was not significantly increased but the average chain length of amylopectin was greater, resulting in an increase in apparent amylose content as judged by iodine binding. In addition, the starch had much higher levels of phosphorous. PMID- 10363369 TI - The HMG-I/Y protein PF1 stimulates binding of the transcriptional activator GT-2 to the PHYA gene promoter. AB - The DNA-binding proteins PF1 and GT-2 are factors that bind to different functionally defined, positively acting cis-elements in the PHYA genes of oat and rice, respectively. PF1 is an HMG-I/Y protein, with its cognate cis-element being an AT-rich sequence, designated PE1, whereas GT-2 is a transcriptional activator with twin DNA binding domains that recognize a triplet of GT-boxes in a complex motif designated GTE. To further define the DNA-binding activity of PF1 and to explore potential inter-relationships between the two factors, we have performed a series of in vitro DNA-binding experiments with both PE1 and GTE target sites. The data show that, consistent with its membership of the HMG-I/Y protein family, PF1 can bend DNA when bound to PE1. In addition, PF1 can bind promiscuously, with varying affinity, to other AT-containing motifs, including GTE. When co-incubated with GT-2, PF1 enhances the specific DNA-binding activity of GT-2 toward GTE, the first report of such activity for a plant HMG-I/Y protein. This enhancement takes place without demonstrable physical contact between the two proteins, suggesting the possibility of a novel, indirect mechanism of recruitment involving DNA target-site pre-conditioning. The evidence indicates therefore that PF1 and GT-2 do not perform functionally equivalent roles in positively regulating oat and rice PHYA gene expression. However, the data suggest the possibility that PF1 may act as an architectural factor, promiscuously recognizing a spectrum of AT containing elements in plant promoters, with the general function of catalyzing enhanced binding of conventional cognate transcriptional regulators to these elements via DNA bending. PMID- 10363370 TI - Biological functions of proline in morphogenesis and osmotolerance revealed in antisense transgenic Arabidopsis thaliana. AB - Many organisms, including higher plants, accumulate free proline (Pro) in response to osmotic stress. Although various studies have focused on the ability of Pro as a compatible osmolyte involved in osmotolerance, its specific role throughout plant growth is still unclear. It has been reported that Pro is synthesized from Glu catalyzed by a key enzyme, delta 1-pyrroline-5-carboxylate synthetase (P5CS), in plants. To elucidate essential roles of Pro, we generated antisense transgenic Arabidopsis plants with a P5CS cDNA. Several transgenics accumulated Pro at a significantly lower level than wild-type plants, providing direct evidence for a key role of P5CS in Pro production in Arabidopsis. These antisense transgenics showed morphological alterations in leaves and a defect in elongation of inflorescences. Furthermore, transgenic leaves were hypersensitive to osmotic stress. Microscopic analysis of transgenic leaves, in which the mutated phenotype clearly occurred, showed morphological abnormalities of epidermal and parenchymatous cells and retardation of differentiation of vascular systems. These phenotypes were suppressed by exogenous L-Pro but not by D-Pro or other Pro analogues. In addition, Pro deficiency did not broadly affect all proteins but specifically affected structural proteins of cell walls in the antisense transgenic plants. These results indicate that Pro is not just an osmoregulator in stressed plants but has a unique function involved in osmotolerance as well as in morphogenesis as a major constituent of cell wall structural proteins in plants. PMID- 10363371 TI - The Arabidopsis flowering-time gene LUMINIDEPENDENS is expressed primarily in regions of cell proliferation and encodes a nuclear protein that regulates LEAFY expression. AB - Mutations in the LUMINIDEPENDENS (LD) gene of Arabidopsis thaliana (L.) Heynh. (Arabidopsis) confer a late-flowering phenotype, indicating that LD normally functions to promote the floral transition. RNA and protein blot analyses, along with the analysis of transgenic plants containing a fusion between a genomic fragment of LD and the reporter gene uidA (GUS), indicate that LD is expressed primarily ipical proliferative regions of the shoot and root, including the shoot apical meristem and leaf primordia. Subcellular localization studies indicate that LD is a nuclear protein, consistent with its previously proposed transcriptional regulatory role. We have also found that in an apetala1 cauliflower (ap1 cal) background the ld mutation converts the reproductive shoot apex to a more vegetative state, a phenotype that is similar to that seen for the leafy (lfy) mutant. Furthermore, in situ hybridization analysis indicates that LFY levels are drastically reduced at the apex of ld ap1 cal plants after bolting. These data are consistent with the idea that at least one function of LD is to participate in the regulation of LFY. PMID- 10363372 TI - Isolation of a gene encoding Arabidopsis membrane-associated acyl-CoA binding protein and immunolocalization of its gene product. AB - Until recently, only cytosolic acyl-CoA binding proteins (ACBPs) have been characterized. The isolation of an Arabidopsis thaliana cDNA encoding a novel membrane-associated ACBP that accumulates in developing seeds, designated ACBP1, has provided evidence for the existence of membrane-associated forms of ACBPs (Chye, 1998, Plant Mol. Biol. 38, 827-838). We now report on the isolation of its corresponding gene from an A. thaliana Columbia genomic library using the ACBP1 cDNA as a hybridization probe. Nucleotide sequence analysis of Arabidopsis ACBP1 showed that its promoter lacks a TATA box, resembling the promoters of rat, Drosophila and human genes encoding cytosolic ACBP and suggesting that it is a housekeeping gene. We show by Western blot analysis that ACBP1 expression in developing seeds coincides with lipid deposition and that homologues of membrane associated ACBP1 exist in other plants. Using light microscopy, we show that ACBP1 is strongly expressed in the embryo at the cotyledons, hypocotyl, procambium of the axis and in most peripheral cells of the cotyledons and hypocotyl. Immunogold labelling localized ACBP1 to vesicles, to the plasma membrane especially at epidermal cells of heart, torpedo and cotyledonary stage embryos, and to the cell wall of the outer integument cells at the seed coat. Our results suggest that ACBP1 is involved in intermembrane lipid transport from the ER via vesicles to the plasma membrane where it could maintain a membrane associated acyl pool; its immunolocalization to the cell wall of outer integument cells at the seed coat suggests a role in cuticle and cutin formation. PMID- 10363373 TI - The TCP domain: a motif found in proteins regulating plant growth and development. AB - The cycloidea (cyc) and teosinte branched 1 (tb1) genes code for structurally related proteins implicated in the evolution of key morphological traits. However, the biochemical function of CYC and TB1 proteins remains to be demonstrated. To address this problem, we have analysed the predicted secondary structure of regions conserved between CYC and TB1, and looked for related proteins of known function. One of the conserved regions is predicted to form a non-canonical basic-Helix-Loop-Helix (bHLP) structure. This domain is also found in two rice DNA-binding proteins, PCF1 and PCF2, where it has been shown to be involved in DNA-binding and dimerization. This indicates that the conserved domain most probably defines a new family of transcription factors, which we have termed the TCP family after its first characterised members (TB1, CYC and PCFs). Other plant proteins of unknown function also belong to this family. We have studied two of these in Arabidopsis and have shown that they are expressed in rapidly growing floral primordia. This, together with the proposed involvement of cyc and tb1 in influencing meristem growth, suggests that many members of the TCP family may affect cell division. Some of these genes may have been recruited during plant evolution to generate new morphological traits. PMID- 10363374 TI - [Current controversies on kidney transplant from living donor]. PMID- 10363375 TI - [Consensus conference on Peyronie's disease. National Meeting of the Andrology Group. Spanish Urology Association. La Coruna, February 27-28, 1998]. PMID- 10363376 TI - [Relationships between diuresis, urine pH and lithogenesis]. AB - Although an increased water intake is suitable in the pathophysiology and treatment of renal lithiasis, less attention is given to changes caused by water diuresis on urinary pH. The objective of this study was to show the relationship between urinary flow and pH. On two consecutive days, at the beginning and end of a four-hour interval starting at 8:00 am, volume and pH in urine, as well as plasma osmolality, haematocrit and protein concentrations were measured; all specimens were collected under normal conditions except for those collected on the second day at noon, which were collected after a water load equivalent to 1.5% body weight taken between 8:00 and 9:00 am. A decrease in serum osmolality, haematocrit and protein concentrations was noted. Urinary pH changes were different based on baseline values. Baseline urine pH values lower than or equal to 5.8 resulted in increased values, while baseline values greater than or equal to 6.5 gave decreased values. Mean increase in pH as a result of greater water intake was 0.57 units. PMID- 10363377 TI - [Upper urothelium tumor in patients treated with radical cystectomy for transitional carcinoma of the bladder]. AB - OBJECTIVE: To evaluate the incidence and characteristics of tumours in the upper endothelium (TUE) that develop in patients with transitional carcinoma of the bladder treated with radical cystectomy. MATERIAL AND METHOD: Between 1986 and 1996, 160 evaluable patients who underwent cystectomy due to transitional cancer of the bladder were reviewed and found to be infiltrant in 96% cases. Follow-up either until death or to the date of the study, was carried out with intravenous urography (IVU) in the first 6 months with additional urographies at least every two years. RESULTS: Five (3.1%) patients showed progress of the upper endothelium tumour, which was multifocal in 3 patients and infiltrant also in 3. No association was seen in these patients with in situ carcinoma of the bladder, or urethral invasion by the primary tumour: only one patient had tumour involvement of end ureters. After three months from diagnosis, tumour-related mortality was 50%. Incidence of upper endothelium tumours in patients with infiltrant tumour of the bladder was lower (1.9%) than in patients with surface tumour of the bladder (16.6%). CONCLUSION: Based on data from our series, the incidence of TUE was 3.1% with a mean time interval between cystectomy and TUE diagnosis of 25.4 months. IVU was diagnostic only in 40% cases. No risk factors were identified in our patients, and mortality due to advanced stage of TUE at three months was 50%. The high percentage of patients with advanced TUE in our series warrants the addition of an annual IVU in the follow-up of these patients. PMID- 10363378 TI - [C.F.C.-type ureterosigmoidostomy. Results]. AB - This paper presents the results obtained with a C.F.C. type ureterosygmoidostomy technique described by us. A total of 25 patients aged between 52 and 74 years (mean age, 65 years), 23 (23/25) male and 2 (2/25) female were evaluated. 24 of the total 25 patients had infiltrant neoplasia of the bladder (pT2: 8; pT3a: 12; pT3b: 4) which was graded as moderately differentiated (G2: 10) or undifferentiated (G3: 14). The remaining patient, a female, had tuberculous microbladder. Male patients underwent radical cystoprostatectomy (23/25); while in two females traditional cystectomy was performed (2/25). Patients with neoplasia of the bladder (24/25) were performed bilateral lymphadenectomy prior to radical surgery. All 25 patients were performed C.F.C. type ureterosygmoidostomy (Actas Urol Esp 20: 324, 1996). Follow-up of patients was carried out over a mean period of 27 months (July 1994-October 1997). The complications reported were 4 cases of ureterointestinal stricture and one stercoral fistula. The strictures were resolved with endoscopic techniques and the fistula through discharge colostomy. Death (6/25) occurred as a result of the tumour progression, and in no case was related to the surgical technique. All patients showed daytime continence (100%), and 22/25 were also continent during the nighttime (88%): there were occasional leaks in 5/25 (20%). No metabolic disorders were seen in any of the patients. (All patients were given drug therapy to prevent metabolic acidosis). PMID- 10363379 TI - [Management of obstructive uropathy in pregnancy]. AB - INTRODUCTION: Obstructive uropathy in pregnant women is a relatively common condition. It can be difficult to assess due to the frequency with which physiologic dilation of the upper urinary tract is seen in pregnant women. PATIENTS AND METHOD: Over the past 3 years 40 pregnant women with symptomatic obstructive uropathy were seen in our service. RESULTS: Most pregnant women responded to conservative treatment (pain killers and antibiotics). In the remaining group, instrumentation was necessary through the urinary route: double J ureteral catheterism (10 patients: 6 due to uterine compression and 4 to lithiasis), percutaneous nephrostomy (4 patients: 2 due to ureteral catheter obstruction impossible to replace, and two due to urinary sepsis), or ureterorenoscopy (1 patient with lithiasis). CONCLUSIONS: The single most common cause for obstructive uropathy in our experience is ureteral compression due to a gravid uterus. Choice therapy in most cases is conservative treatment. When in spite of conservative treatment signs and symptoms persist, urinary by-pass with antibiotic prophylaxis must be performed. Ureterorenoscpy as a diagnostic and therapeutical approach should be taken into consideration in pregnant women with ureteral lithiasis. Incidence of pre-term labour was not higher than usual. PMID- 10363380 TI - [Economic impact of complications after transurethral resection of the prostate on its cost throughout the process]. AB - INTRODUCTION: Considering that BPH is a highly common condition in the urological practice, that demographic changes further emphasise this feature and that the analysis of costs, funding and health care management are becoming increasingly under scrutiny, it is important to know the cost per procedure of the most commonly used therapeutical approaches and the impact of the complications on such cost. OBJECTIVES: 1. To know the cost per procedure of prostate TUR as PBH treatment. 2. To know the percentage of complications involved and the way in which they impinge on cost. MATERIAL AND METHOD: Considering all data gathered from the annual surgical activity in our service with regard to this procedure and the percentage of complications after studying 105 cases over an evolution period of 6 to 50 months, the Planning and Management control Unit at the H.C.S. reported on the expenditure generated by this activity. RESULTS: The cost per procedure in prostate TUR due to BPH in our environment is 191.030 pesetas at 1997 value. Morbidity affects 23% patients. Diagnosis and treatment of complications increases the overall cost by 7% thus reaching 205.096 pesetas. CONCLUSIONS: The knowledge of the above results provides important data on the expenditure generated by the activity of our Service for funding and management purposes. It also allows us to perceive therapeutical results as a measure of quality control, and to redefine those actions that will allow us to improve our results. PMID- 10363381 TI - [ESWL-resistant lithiasis]. AB - INTRODUCTION: To the extent in which the "lithotripter" improves technically. SWEL experts provide different explanations to the failures of this technique. It will depend on the type of "lithotripter" as well as the calculus and its features (size, number, location, composition, obstruction, impact, absence of expansion chamber, presence of ureteral catheter, ...). Not all facts in SWEL have a clear explanation today. Physically, the "cavitation" phenomena (shock, rebound, negative pressure, explosion, heat, ...) explain almost anything that takes place during SWEL. Certainly, the type of lithotripter has some influence, but the calculus fragility, determined by the chemical composition and the crystalline architecture, could be more determinant. MATERIAL AND METHOD: From a total series of 6,500 SWEL procedures performed in the Lithiasis-Lithotrity Unit at the "Jimenez Diaz" Foundation (JD) (January 1991-July 1998), 20 cases considered as failures after multiple SWELs were analyzed. Also the actual diagnostic tests (X-rays, helicoid CAT, densitometry, ...) were studied to establish a pre-SWEL chemical recognition of the calculi that may determine the behaviour of each case prior to treatment. RESULTS: After multiple SWELs (average 5 sessions) negative results were obtained in 65% cases. These cases were resolved with surgery (38%), ureterorenoscopy and ultrasound lithotrity (23%) or watchful wait in absence of signs and symptoms (39%). 57% were calcium phosphate calculi, 29% calcium oxalate monohydrate (COM) and 14% hypercalciuria calculi. CONCLUSIONS: SWEL resistant cases, either unresolved or undergoing multiple SWELs, demonstrate the existence of calculi that cannot be broken by SWEL, although no coincident or similar reasons can be established in all cases. Calcium phosphate dihydrate (brushite) and calcium oxalate monohydrate (COM) together with cystine are the most difficult to destroy with current shockwaves. Helicoid CAT could recognise chemically each case prior to SWEL, since it basically differentiates the most frequent ones, uric acid, struvite and calcium oxalate. PMID- 10363383 TI - [Hemangioma of the glans penis]. AB - The haemangiomas are benign vascular tumors of frequent apparition in infant, but not very much described in penis glans in the literature. We present a case with location in penis glans, in a short age male, that it was treated through conventional surgery with good results. We accomplish a review of this pathology and we make reference to the different ways of treatment today. PMID- 10363382 TI - [Histologic-ultrasonographic correlation of gland asymmetry and "pseudoruptures" of the prostatic capsule]. AB - An histo-ultrasound correlation was carried out between the information obtained with pre-operative transrectal ultrasound and that obtained with the histopathological study of 29 prostate specimens from patients with mean age 63 years (range, 52 to 71 years), who underwent radical cystoprostatectomy for infiltrant neoplasia of the bladder (22/29) and radical prostatectomy for prostate cancer (7/29). The (extrinsic and intrinsic) ultrasound parameters were analyzed focusing in the study of the prostatic capsule or "pseudocapsule". With this comparative, echographic and histological study the concept of capsular "pseudorupture" that results from the presence of (arterial and/or venous) vessels, nerves and fibromuscular folds at the periprostatic fat tissue is introduced. The interruption of the capsular echogram was seen as an indication of tumoral involvement of the capsule, and extracapsular spread. Our results allow us to suggest that this assertion should be reconsidered since these capsular "interruptions", "irregularities" or "pseudoruptures" are present in the normal prostate. Similarly, glandular asymmetry in relation with an heterogenous growth in one lobe in defined to differentiate it from that seen in neoplastic processes. PMID- 10363384 TI - [Retroperitoneal germ cell tumor]. AB - We report a case of retroperitoneal extragonadal germ-cell tumor in an 17 years old patient who presented with aedema and pain in left inferior extremity asociated with hemopthysis caused by pulmonar metastasis, who was treated with chemotherapy and resection of residual mass and pulmonary nodes. Dyagnosis was stableshed by fine neadle aspiration biopsy of the wass. We comment on the difficult of stableshing differential dyagnosis between retroperitoneal extragonadal germ-cell tumor and metastasis of a testicular tumor. Dyagnosis is stableshed by the finding of a histologically malignant germ-cell tumor with normal testis. We considered physical examination and ecographyc exploration enough for a correct dyagnosis. PMID- 10363385 TI - [Leydig cell tumor: report of 2 cases]. AB - Tumors from the gonadal stroma represent 4% among testicular tumors. Leydig cell tumors are the most common neoplasms among them and account for 1-3% of all testicular tumors. Two cases of testicular Leydig cell tumors in adult patients are presented. Presentation, diagnosis and therapeutic aspects are discussed. Both were treated with radical orchiectomy through an inguinal approach. Cas 1 was diagnosed in a cryptorchid testis and developed hepatic metastasis that were successfully treated with chemotherapy. Cas 2 was incidentally diagnosed on ultrasound. Both of them remain alive and free of disease. PMID- 10363386 TI - [Penile injuries: our experience]. AB - Trauma of the penis is relatively uncommon. Not many centres and even less urologists have extensive experience in the comprehensive management of this type of injury. Revision of the series accrued in our hospital over 20 years, in each case explaining the mechanism of occurrence, treatment applied and results obtained. PMID- 10363387 TI - [Pelvic lipomatosis: a case report with diagnostic and therapeutic approach]. AB - Pelvic lipomatosis is a rare disease of unknown etiology characterized by benign proliferation of fat in the pelvis. We describe a 27-year-old man with pelvic and retroperitoneal lipomatosis causing a severe urinary and fecal obstruction. The diagnosis was evaluated by barium enema, intravenous urogram, computerized tomography and magnetic resonance imaging. The therapeutic approach is described and discussed. PMID- 10363388 TI - [Obstructive uropathy secondary to retrovesical hydatid cyst]. AB - The genitourinary affectation by the hydatidosis, is the third in frequency after the hepatic and pulmonary affectation. The finding of an obstructive uropathy by a retrovesical hydatid cyst is uncommon, but it must be in account in high incidence zones. We present a case of this pathology, accomplishing a review of their etiopathogenic mechanisms, as well as of their arrival way, diagnostic approach and treatment. PMID- 10363389 TI - [Urology images. Cutaneous lymphoma in the perineal scrotum]. PMID- 10363390 TI - [Reflections on self-management. 2 years after its start]. PMID- 10363391 TI - [A descriptive study of community-acquired pneumonia in childhood. A primary care perspective]. AB - OBJECTIVES: MAIN OBJECTIVE: To determine the incidence of child pneumonia in our health district. SECONDARY OBJECTIVES: to establish its clinical and epidemiological characteristics, to establish the percentage resolution of the illness within primary care (PC), to describe the treatments given, and to compare admission rates according to whether the first consultation was in PC or hospital casualty. DESIGN: A retrospective, longitudinal, descriptive study. SETTING: Primary care. PARTICIPANTS: 63 episodes diagnosed between 30-8-96 and 1 9-97 in 1604 children under 15. MEASUREMENTS: Statistical measurements included: 95% confidence intervals, the ji squared test, Fisher's exact test. RESULTS: Incidence was 3.9% (CI: 3-4.9%); female predominance, 53.3% (CI: 40.7-66%); seasonal predominance, autumn-winter 64.5% (CI: 52.6-76.4%). The most common signs were: high temperature, 93.3% (CI: 87-99.6%); cough, 66.1% (CI: 54-78.2%). 90.5% (CI: 83.2-97.7%) of pneumonia cases treated in PC evolved satisfactorily. 9.5% (CI: 2.3-16.8%) needed hospital admission. Patients who attended hospital on their own initiative went into hospital more often than those who attended the PC paediatrics clinic as their first option (p = 0.002). CONCLUSIONS: The incidence of pneumonia among children is lower than in other countries. The majority are treated and resolved properly in PC. Analytic studies are needed to determine whether first attending a hospital casualty department instead of a PC paediatric clinic involves a greater risk of hospital admission. PMID- 10363392 TI - [The lipid profile in the children and adolescents of Caceres Province]. AB - OBJECTIVE: To analyse the lipid profile of the infant and young population of our province, and to compare our findings with those of other similar Spanish studies. DESIGN: A cross-sectional descriptive study. SETTING: School population from 2 to 16 years old in the province of Caceres (N = 91083). PARTICIPANTS: A representative and proportional sample of 2150 children, with the two sexes at 50%. INTERVENTIONS: The total cholesterol (TC), HDL-C (enzymatic technique) and LDL-C (Friedewald formula) were determined. RESULTS: The mean values of TC, LDL-C and HDL-C (mean +/- SD) were 182.77 +/- 28.96, 114.19 +/- 26.54 and 57.31 +/- 13.97, respectively. Comparison of our lipid figures by age and sex with those of other Spanish studies (Fuenlabrada, Nino Jesus, Navarra and RICARDIN) and with the North American LRCP showed, in general, higher means of TC and LDL-C and/or lesser HDL-C levels in our population. CONCLUSIONS: Comparison with earlier studies shows that children from Caceres have a more atherogenic lipid profile. Social and methodological differences may have affected the results. However, we think that, given the time elapsed since previous studies, the differences found may reflect progressive changes in the Spanish diet towards higher consumption of saturated fats. PMID- 10363394 TI - [The impact of an intervention strategy in the prescription of generic drugs in a primary care area]. AB - OBJECTIVE: To examine the effects of specific interventions on generic prescribing in general practices. DESIGN: Non randomized controlled study. SETTING: General practices in 2 health areas of Madrid. PARTICIPANTS: 5 general practices (intervention group) and 5 general practices (control group). INTERVENTIONS: In 1994, intervention group received monthly educational seminars on generic drugs and computer-produced prescribing feedback. The control group received not seminars and not feedback. Prescription monitoring of both groups continued during 1994. MEASUREMENTS AND MAIN RESULTS: Area factor before and during intervention, and intervention factor for both areas (control and intervention) related with prescribing volume and costs of generics were analysis. Prescribing volume of generics in the intervention group was significantly greater than for control group (p < 0.01) by both area and intervention factors. Prescribing costs of generics in the intervention group was significantly greater for control group (p < 0.05) by area factor but both groups were significantly different by intervention factor. CONCLUSION: Educational seminars and feedback information on generics improve generic prescribing but it should be evaluated for broader areas of physicians on prescribing costs. PMID- 10363393 TI - [An episode of the prevention of cardiovascular risk factors by age and sex in the Community of Valencia]. AB - OBJECTIVE: To typify the episodes of early detection of cardiovascular risk factors (CRF) and to calculate their frequency by age and sex groups. DESIGN: An observational, prospective and multi-centred study. SETTING: Twenty health centres distributed in the three provinces of the Community of Valencia. PATIENTS: Episodes of prevention of CRF in users of both sexes aged between 18 and 64 who, in the three years prior to the study, had undergone no CRF early diagnosis protocol. They were included by means of consecutive proposal with informed consent. MEASUREMENTS AND RESULTS: The diagnostic protocol of the Plan for Prevention of Cardiovascular Diseases (PPCVD) was applied. The variables analysed were: age, sex, number and duration of consultations, CRF diagnosed previously, and CRF diagnosed at the intervention. 632 episodes were analysed, with a mean 1.44 (CI: 1.39-1.49) consultations per episode and a mean duration of 10 minutes 53 seconds. At the start of the study 60.3% of the population did not have CRF, but after the intervention only 17.2% had no CRF diagnosed. CRF frequency after the intervention was: diabetes 4.5% (CI: 3.2-12.2), alcohol consumption 5.5% (CI: 2.2-13.1), hypertension 14.9% (CI: 7.7-22.2), obesity 30.8% (CI: 24.3-37.3), tobacco habit 33.2% (CI: 26.8-39.6), lipaemia 42.5% (CI: 36.6 48.5), and sedentary life-style 54.9% (CI: 49.6-60.2). CONCLUSIONS: CRF diagnosed most often after the intervention were: sedentary life-style, tobacco habit and obesity; and the least commonly diagnosed was alcohol consumption. The application of the PPCVD protocol was most effective in the youngest age-groups and women. The "episode" as a unit of analysis is a useful and feasible instrument for investigating the procedures and results of primary care preventive activities. PMID- 10363395 TI - [Functional stabilization versus orthopedic immobility in grade-I-II (mild) ankle sprain]. AB - OBJECTIVE: To proof that functional treatment is better than the orthopedic one in the slight ankle sprain. DESIGN: Clinical trial. SETTING: A primary care center, a outpatient clinic and a teaching hospital in Malaga. PATIENTS: 80 patients treated in emergence room for ankle ligament injuries. They were systematically sampled, 40 in each group of treatment. INTERVENTIONS: Functional stabilization or elastic adhesive bandaging during 7 days without support. Immobilization after the acute phase permitting then the support. Orthopedic stabilization or subsequent ferule during 21 days, followed by ankle rehabilitation. MEASUREMENTS AND MAIN RESULTS: We compared the healing time, treatment acceptance, absent of work time and need of rehabilitation in both groups. Functional stability is more effective than the orthopedic one: 1.39 times better in terms of curing, 7 times in terms of personal hygiene, 4.25 times in terms of need of rest, 5.5 times better in terms of need of rehabilitation. CONCLUSIONS: We confirm the initial hypothesis of the advantage of the functional treatment. PMID- 10363397 TI - [An evaluation of the feasibility and validity of a scale of social assessment of the elderly]. AB - OBJECTIVES: To analyse the reliability and validity of a scale of social evaluation of the elderly. DESIGN: Descriptive, cross-sectional study. SETTING: Primary care. PATIENTS: Sample of 1062 people from the over-65 population. INTERVENTIONS: The scale evaluated has five items (family situation, economic situation, housing, relationships and social support), and an overall score is obtained. Its reliability was evaluated by an interview with two observers, and validity by contrasting the score obtained on the scale with a reference criterion of an independent, blind assessment by a social work expert. MEASUREMENTS AND MAIN RESULTS: The intraclass correlation coefficient (inter observer reliability) was 0.957. The Cronbach alpha coefficient was 0.4467, which denoted moderate to low internal consistency. Sensitivity and specificity were calculated for the validity of the criterion. Nevertheless, to detect social problems in care practice, probability proportions for different levels on the scale were more useful. These ranged from 1 to 23, while in the detection of social risk they ranged from 1 to infinity. CONCLUSIONS: The scale studied by us as a measuring instrument enables risk situations and social problems to be detected with good reliability and acceptable validity. It should be introduced into the care practice of professionals working in the social or health care of the elderly. PMID- 10363396 TI - [The prevalence of antibodies to the rubella virus in pregnant women at a health center]. AB - OBJECTIVE: To know the prevalence of antibodies against rubella in pregnant women of a health centre. DESIGN: Cross-sectional descriptive study, for 4 years. SETTING: Lliria health centre. Zone 9. Area 5. Valencia. PATIENTS: Random sample of 405 pregnant women (15-45 years) of 3500 women in fertile age, extracted from the obstetrics unity. MEASUREMENTS AND RESULTS: We collected the following variable: age of the mother in the last gestation, number of previous gestations (including abortions), determination of antibodies against rubella, degree of schooling and occupation index. The results show a prevalence of antibodies against rubella of the 95.2% (CI 95% 93.1-97.3). We find meaningful differences between the antibodies protective titles and the age (p < 0.0005) and instruction level (p = 0.0263), so when the women are elder and they have a instruction level better the antibodies protective titles are bigger. We don't find meaningful differences with the mother occupational level (p = 0.0945), either with the number of previous gestations (p = 0.2947). The prevalence of antibodies against rubella, according to groups of age in the pregnant women population, varies from 81.8% (CI 95% 59.7-94.8) in the group of 15 to 19 years, until a 100% (CI 95% 66.4-100) in the group of 40-44 years. The relationship between the number of gestations and the lack immunization against rubella is for the first pregnancy 52.6%, second pregnancy 36.8%, several pregnancy 10.5%. CONCLUSIONS: The prevalence of antibodies against rubella in pregnant women is 95%, similar to other published studies. This prevalence increases with the age what suggests the primary paper of the wild virus in the maintenance of the rate of prevalence. It should be to incise more in the immunization after the labour, overcoat, in those pregnant women with less schooling, so corroborate our results. It is necessary to take the adequate measures so that 47.4% of the total of seronegative women (with more than one pregnancy), don't escape to the controls of the sanitary system. PMID- 10363398 TI - [Omeprazole and gastric protection]. PMID- 10363399 TI - [Premalignant lesions of the oral cavity in primary care]. PMID- 10363400 TI - [The checking of the vial-shaking test for the detection of the freezing of toxoid-based vaccines]. PMID- 10363401 TI - [Bromocriptine for the suppression of lactation and severe hypotension]. PMID- 10363402 TI - Intrapericardial caval injury due to blunt trauma. AB - BACKGROUND: Report of diagnosis and treatment of intrapericardial vena caval injury caused by blunt thoracic trauma, an unusual cause of cardiac tamponade. METHODS: A 43-year-old male motor vehicle accident victim suffered a lacerated intrapericardial inferior vena cava leading to cardiac tamponade. Positive clinical findings were hypotension and tachycardia without indication of external chest trauma. RESULTS: Abdominal computed tomography was negative, but ultrasound demonstrated cardiac tamponade and fluid in the abdomen. Pericardiocentesis was performed, and nonclotted blood was aspirated. Laparotomy showed intra-abdominal blood and splenic capsule avulsion. Sternotomy revealed a laceration of the inferior vena cava, which was repaired. CONCLUSIONS: Signs of cardiac tamponade and a history of blunt thoracic trauma caused by deceleration injury suggests intrapericardial inferior vena cava injury. Median sternotomy is the optimal choice for caval repair. PMID- 10363403 TI - Left atrial myxomas presenting with right hand weakness. AB - A 63-year-old man with multiple left atrial myxomas presenting with right hand weakness is described. Two-dimensional echocardiography obtained during the evaluation for his stroke showed a large left atrial mass. At surgery, two separate left atrial myxomas were excised. Pathology confirmed the diagnosis. Although left atrial myxomas are uncommon, they should be included in the differential diagnosis of stroke, especially in patients who present without cardiovascular or cerebrovascular risk factors. The absence of cardiac symptoms and signs does not rule out a cardiac myxoma. PMID- 10363404 TI - Clinical and laboratory evolution of a culture-confirmed case of human granulocytic ehrlichiosis. AB - A 74-year-old man from suburban New York City, who was hospitalized because of chest pain and fever, was diagnosed as having human granulocytic ehrlichiosis on the eighth hospital day. Although leukocyte and platelet counts were normal on admission, they fell to abnormally low values then normalized prior to specific therapy against the human granulocytic ehrlichiosis agent. Intracytoplasmic inclusions suggestive of Ehrlichia were observed in up to six percent of granulocytes, and the human granulocytic ehrlichiosis bacterium was cultured in an HL 60 human promyelocytic cell line. The patient improved dramatically within 24 hours of doxycycline treatment, after failing to improve on various beta lactam antimicrobial agents. He was discharged from the hospital 14 days after admission. Because human granulocytic ehrlichiosis was not diagnosed until his eight hospital day, clinical and laboratory parameters prior to specific treatment were available. This case illustrates the clinical and laboratory evolution of the infection with human granulocytic ehrlichiosis agent in humans. PMID- 10363405 TI - Community acquired Pseudomonas aeruginosa pneumonia. AB - Pseudomonas aeruginosa is an uncommon cause of community acquired pneumonia in immunocompetent hosts. We report two cases that did well once appropriate and prolonged antimicrobial therapy was initiated. They had no evidence of immune deficiency. The initial consideration was pulmonary tuberculosis in both cases given the subacute presentation, significant weight loss, and findings on chest roentgenogram. PMID- 10363406 TI - Continuous infusion of beta-lactam antibiotics. PMID- 10363407 TI - Fraud bounties bad idea for Medicare. OP-ED published in the New Haven Register, 17 March 1999. PMID- 10363408 TI - An ethical dilemma. PMID- 10363409 TI - Adult onset Still's disease presenting as aseptic meningitis in a young healthy female. AB - A 20-year-old white female presented with symptoms of upper respiratory tract infection, meningeal signs, rash, and fever. Initial laboratory data revealed a leukocytosis and abnormal CSF. An initial working diagnosis of the Aseptic Meningitis Syndrome was made. She did not respond to antimicrobial therapy. All culture results and viral titers were negative. One week into her hospital course, the diagnosis of Adult Onset Still's Disease (AOSD) was made. Antibiotics were discontinued and nonsteroidal anti-inflammatory drugs (NSAIDS) begun. The patient showed marked improvement within 24 hours. This case reveals that AOSD is an important consideration in the differential diagnosis of Aseptic Meningitis. Meningeal signs and abnormal cerebrospinal fluid (CSF), both detected in this patient, are very rare occurrences in Still's disease. PMID- 10363410 TI - Medical text books on the World Wide Web. AB - The availability of medical textbooks for physicians on the web is rapidly increasing. Internet sites that maintain a listing of such resources are briefly described, and descriptions of particularly valuable fee based and free medical textbooks web sites are categorized. PMID- 10363411 TI - Radical or conservative? PMID- 10363412 TI - [Management of adnexal masses in gynecology using conservative measures. Is the laparoscopic approach convenient?]. PMID- 10363413 TI - [Struma ovarii: a variety of monodermic teratoma of the ovary. Report of 8 cases]. AB - Inside the germinal neoplasms of the ovary, the monodermic or specializad teratomas constitute a not very frequent variety. The struma ovarii represents 2.7% as the form more common of this teratomas type, constituted by thyroid tissue. Struma ovarii can associate to other tumors of the ovary of germinal type as dermoid cyst, carcinoid tumor or non germinal tumors as type as dermoid cyst, carcinoid tumor or non germinal tumors as mucinous or serous cyst or Brenner tumor. Most of the struma ovarii are benign and 5-10% only are malignant, of these group 40% has only associated to extraovaric dissemination. The objective of the present work is to analyze the clinical, epidemical and biological characteristics of the struma ovarii. Eight cases of struma ovarii are presented, that occupy 4% inside the ovary tumors diagnosed and treated in our hospital. The clinical charac-teristics of these tumors were: age average 45.5 tears, pelvic mass accompanied by distension and abdominal pain. They were managed as anyone ovary tumor, in not any case the diagnosis it was suspected, 4 were presented of the left side and 4 in the right, the weight average was 726.2 g. Only in 4 cases was carried out the transoperative diagnosis and the eight cases confirmed in the postoperative. In not any case hyperthyroid symptoms, neither morphological criteria of malignancy were presented. Only in two cases the struma ovarii was reported as pure, in the 6 remaining it is accompanied with another type of ovarian neoplasm. PMID- 10363414 TI - [Recurrent fetal loss. A multifactorial problem?]. AB - The fetal recurrent loss (P.F.R.), it is a clinical disturbance associated with multiple factors, that it are in a frustrating situation so much for the couples like for the doctor. The objective was to evaluate 39 couples with P.F.R. in order to establish the responsibility of each factor and proceed to treatment I specify, considering as success the get a pregnancy with viable product. The protocol included clinical history, histerosalpingography, ultrasound, analysis cromosomal, antibodies for TORCH and antiphospholipids, test endocrine specifies, genitals cultivation and biopsy of endometrial. 23% it of the fetal losses is due to only factor; 64.2% it is due to multiple factors and 12.8% they don't have apparent factor. The infectious factors, endocrine, anatomical and autoimmunity was the more constants. Pregnancy with viable product in the 82% was achieved of the cases. We concluded that the P.F.R. it is a problem that is due to multiple factors and that it require a diagnosis-therapeutic integral focus. PMID- 10363415 TI - [Vaginally assisted laparoscopic hysterectomy vs. abdominal hysterectomy. Initial experience]. AB - A longitudinal prospective study was performed including 22 patients that were submitted to hysterectomy, for diverse being uterine pathologies, between March 1st. 1995 and until August 31st. 1996, in private practice at Mexico City. The patients were divided into two groups: I) Patients submitted to total abdominal hysterectomy (TAH), 10 patients, and II) Patients submitted to vaginally assisted laparoscopical hysterectomy (VALH), 12 patients. The inclusion criteria for each group depended on the patient's weight: those with an overweight of more than 20% were included in group I, and those with an overweight of less than 20% were included in group II. All the patients were comparable in age, gestational history and surgical abdominal history. In our experience those submitted to TAH (4.2 weeks) or those submitted to VALH converted into laparotomy (3.5 weeks). In group I three patients presented minor complications (postoperative hemoglobin of less than 10 g/dl--two cases--and febrile morbidity secondary to minor lung athelectasis--one case--) that were solved with conventional measures. The mean surgical time was similar between groups I and II (159 and 152 minutes respectively) and longer in the VALH converted into laparotomy (240 minutes). The mean hospital cost was similar for groups I and II (18.6 thousand pesos). The hospital stay was similar in all cases (3.6, 3 and 3.5 days for groups I, II and the VALH converted cases, respectively). Those patients submitted to VALH electively stayed in the hospital for three day, even though their postsurgical progress was evident, so they could have been discharged of the hospital 24 hours after the surgery. PMID- 10363416 TI - [Heterotopic pregnancy with intrauterine dizygotic twins following embryo transfer in the blastocyst phase]. AB - Ectopic pregnancy is a common complication of in vitro fertilization and embryo transfer (IVF-ET). On other hand, heterotopic pregnancy complicates 1-2% of all IVF-ET pregnancies. Tubal damage as reason for treatment and multiple embryo transfer might predispose patients to this complication. We present a successful treated case of an infertile patient that developed simultaneous twin intra- and single extra- uterine pregnancy after blastocyst-stage embryo transfer. In IVF-ET patients presence of an intrauterine gestation not exclude the possibility of a concomitant extrauterine pregnancy. Awareness of the possibility of heterotopic pregnancy after IVF-ET plays an important role in the successful treatment of this reproductive complication. Transfer of good quality embryos can be a risk factor to develop heterotopic pregnancy. PMID- 10363417 TI - [Dilated myocardiopathy and pregnancy. Report of 3 cases and review of the literature]. AB - Pregnancy in association with dilated cardimyopathy is considered to be a rare entity. The incidence is unknown. The dilated cardiomyopathies (DCM) are defined as diseases of the cardiac muscle without a known cause. Three pregnancies with DCM are reported. In this article the possible hazzards of pregnancy and DCM are discussed, including a review of the literature concerning to diagnosis and management is present. PMID- 10363418 TI - [Changes in the thrombophilic status in patients with pre-eclampsia]. AB - The object of this study was to evaluate the changes in fibrinolysis and clotting inhibitors in patients with preeclampsia and to describe the connection between preeclampsia and blood pressure values. Two groups of pregnant women were prospectively studied at delivery: group 1 women without preeclampsia and group 2 patients with preeclampsia. The variables that were registered are: diastolic blood pressure (DBP), systolic blood pressure (SBP), mean blood pressure (MBP), hemoglobin (Hb), platelet count (Plt), lupus like inhibitor, anticardiolipin antibodies (ACA), antinuclear antibodies (ANA), fibronectina, D dimer, protein S (PS), protein C (PC) and vo Willebrand factor (vWF). 62 pregnant women were included. The patients of group 2 presented high values of Hb (p 0.01), fibronectin (p 0.0001), D-dimer (p 0.01) and lower PC (p 0.04). We found an association between fibronectin and higher values of SBP, DBP, MBP and Hb (p 0.0007) versus lower values of VFW and PC (p 0.002). The low values of total PS were associated with high D-dimer and SBP results (p 0.04 and 0.002 respectively). All patients were ACA/ANA negative. In preclampsia there is a increased hemoconcentration and drop in clotting inhibitors (PC), without fibrinolytic compensatory response (lower D-dimer) and remarked vasopressive effect (hig fibronectin). This changes depend on the stratification of blood pressure. Th SBP and MBP values depend on the haemodynamic changes (Hb, fibronectin), while the increase in DBP expresses a non compensated thrombophilic state. PMID- 10363419 TI - [beta-Endorphins and the male reproductive system]. AB - In the antique Chinese culture, it was already known about opioide for therapeutic usage. On 1975, Hughes et al., identificated two endogen pentapetides with potent opiate activity Leu-enkephalin and Met-enkephalin, ever since beta Endorphin have been considered involucrated in many biological functions. beta Endorphin have found in testis, seminal vesicle and prostate in different species. It has been observed a paracrine effect between in Leydig and Sertoli cells and inhibit the basal release of LH in man by inhibiting its pulsatile discharge. In this paper is discussed, in a general way, localization and production sites of beta-Endorphin especially their participation in male reproductive tract-physiology. Finally, it is mentioned the experimental methodology to identify and quantify beta-Endorphin in blood plasma and semen. PMID- 10363420 TI - [Anion gap in perinatal asphyxia]. AB - The aim of this study was to assess the usefulness of the anion gap (AG) for the diagnosis of Perinatal Asphyxia (PA), as well ass to evaluate the relation of AG with umbilical artery cord pH and the outcome of the newborn. We studied 62 infants in whom umbilical artery blood gases were obtained within the first 15 minutes of life. Serum sodium, chloride and potassium were simultaneously measured. AG was calculated according to the following formula: (Na + K) - (Cl + HCO3). Patients were divided in two groups depending on their pH values. Infants with a pH of 7.10 or less, were assigned to the PA group (n = 22); infants with a cord pH higher than this value, were taken as a control group. Additionally, the total group of patients was divided in survivors and non-survivors. Statistical analysis was done with mean comparation tests. Pearson's correlation index, and the sensibility, specificity and predictive values, positive and negative were calculated for a diagnosis test. Both groups were similar in gestational age, birthweight and serum electrolytes. There was significative difference in the Apgar score at one (4 vs 5.5 for PA and control group respectively) and five minutes (6 vs 7.5 respectively), as well as in the pH (6.89 +/- 0.20 vs 7.25 +/- 0.06) and the anion gap (23.4 +/- 8.1 for PA group vs 13.9 +/- 3.1 in control group). 20 mEq/L was considered as the cut-off point for AG normality. The sensibility of the test for diagnosis PA was 0.81, specificity 0.52, positive predictive value 0.48 and negative predictive value 0.84. From 37 infant newborns with a higher value than 20 mEq/L, were 11 nonsurvivors, than from 25 with a lowest measurement was 4 nonsurvivors, without statistically difference between these groups. From the total of 62 patients, were 47 survivors and 15 nonsurvivors, without difference between them in their AG, Apgar score, gestational age or serum ions. We did find statistically significant differences regarding birthweight (p < 0.05), pH (p < 0.05), and bicarbonate (p < 0.001), all these values were higher in the survivors infants. No significant correlation was found between AG and pH, however was positive correlation between AG and HCO3 (r = 0.33, p < 0.01), sodium (r = 0.22, p < 0.01) and chloride (r = 0.33, p < 0.001). PMID- 10363421 TI - [8th National Congress of Gynecology and Obstetrics 26-30 October 1998. Conference on "Philosophy, Bioethics and the Gynecologist"]. PMID- 10363422 TI - [Response of fetal macrophages in the placenta of pregnant HIV-positive patients with and without antiretroviral treatment]. AB - We studied the cellular answer of placentary macrophages in pregnant women seropositive to the virus of human immunodefficiency (VIH-1) treated with zidovudina (AZT) and didanosine (ddl). Twenty eight pregnant women were studies; there were four groups of seven patients each: The control group; the group with seropositive women without treatment; the group given AZT, and the group that recieved AZT and ddl. Placentary specimens were obtained immediately after delivery. One hundred and fifty chorionic vellosities of cells. The control group showed an average of 26 Hofbauer cells; the seropositive women without antiretroviral treatment, was 115; the patients who received only AZT, the average was 65; and the ones who received a combine therapy AZT and ddl, cellular average was 44. There were no differences in the weight of the products in all the groups, nor congenital malformations in the newborns. The use of medication antiretroviral suppress viral replication, and so, there is a significant answer in the amount and size of Hofbauer cells. The administration of two medicaments is more effective in the cellular immune answer. PMID- 10363423 TI - [Balance of the TH1/TH2 immune response in the materno-fetal interphase in normal and preterm pregnancy. Preliminary results]. AB - The subpopulations don's speak directly of the answers Th1 or Th2 in the compartment, as the are only reflexion of prevalent citocines in the microenvironment: However. It is important its exploration to the taken as starting point of this project. PMID- 10363424 TI - [Embryonic quality in the use of urofolitropin vs recombinant FSH for in vitro fertilization]. AB - There were no differences in both groups as tho the age of the patients; received doses of both types of FSH, nor HMG; but there was as to the amount of captured ovocytes, amount, and quality, embrionary, in special 1+ 2+ in favor of the group that received urofolitropine, specially under 35 years of age. In this study there was better qualy and amount, embrionary, obtained with the use of urofolitropine, as compared with FSH recombinant for in vitro fertilization. PMID- 10363425 TI - [Determination of the tumor necrosis factor in the peritoneal fluid of gynecologic patients with intraperitoneal infections and endometriosis]. AB - Tumor necrosis factor (TNF-alpha) is a cytokine which can be found in peritoneal fluid (PF) of patients with endometriosis and pelvic inflammatory disease (PID) as a response of inflammatory disorder and infections diseases. The cytotoxic effect of this cytokine could be participating in the pathology of different gynecologic problem and be accountable of the high immunological response and damage on the tubal epithelium. The objective of this study was determinate the presence of TNF-alpha in PF of endometriosis patients, fallopian tube occlusion (FTO) and PID and their correlation with different isolated bacteria. Ten mililiter PF were collected and cultured in antificial medium and Mc Coy culture cells for isolation of acrobic, and anaerobic bacteria and Chlamydia trachomatis from 73 patients by laparoscopy. The TNF-alpha activity was determined by L-929 cells endometriosis, 30 PID and 4 had miomas and adherences. The 50.7% of patients were cultive positive, fom these, 31.5% were PID. Chlamydia trachomatis (16%) was the most frecuenty isolated bacteria in these patients. 59.4% of FTO patients displayed TNF-alpha activity. However, only 4% showed positive isolation, in conclusion the detection of TNF-alpha could be useful in active infectious and inflammatory diseases in patients which not present simptomatologic characteristic of these illnesses and plus being attended at for sterility clinical as a result of their incapacity to get pregnant. PMID- 10363426 TI - [Osteoporosis in Mexican postmenopausal women. Magnitude of the problem. Multicenter study]. AB - Incidencia of osteoporosis induced fractures increases with age; risk increases exponentially as bone mass decreases. Women are prone to osteoporosis 2 to 3 times more than men, due to lower "peak" bone mass and the accelerated loss that occurs after the menopause. The prevalence of osteoporosis in with Caucasian postmenopausal women varies from 16 to 30% depending upon the number of sites measured (lumbar spine/hip and/or forearm). The aim of this study was to estimate the prevalence of osteopenia and osteoporosis in pre and postmenopausal Mexican women in relation to men, and to that reported in Caucasia women. The study involved 4,821 apparently healthy subjects (without known risk factors for osteoporosis), 4,467 females and 354 males from 11 different centers of Mexican Republic, 20 to 90 years old, using DXA bone densitometry of lumbar spine and hip. Prevalence of osteoporosis in women is twice that in men (P < 0.001), and it increases with age, particularly after the menopause (P < 0.0001). Our study found a prevalence of osteoporosis of the lumbar spine and/or hip in apparently healthy postmenopausal Mexican women over 50 years of age of 16%, increasing to 20% in those women with or without risk factors who attended voluntarily or by suggestion of their physician to be studied. The prevalence values obtained seem to be lower than those reported for Caucasian women, 30%. We found a prevalence of osteoporosis of 16% and of osteopenia of 57% in women 50 years of age and older. We also found a higher prevalence of osteoporosis in women of the south east part of the county. PMID- 10363428 TI - The roles of athletic trainers and physical therapists in sports medicine. PMID- 10363427 TI - [Initial experience with a minidose of methotrexate in the management of unruptured ectopic pregnancy]. AB - Methotrexate has been used in the conservative management of ectopic pregnancy with good results. Due to its great afinity for the trophoblast it was decided to try unique doses of 50 mg i.m. independently of the body surface. Fifteen patients were studied with ectopic pregnancy by ultrasound and hCG in series that didn't require confirmatory laparoscopy. The ectopic pregnancies broken and/or decompensated were discarded. Average gestational age was 6.1 weeks; the maximal dimention of the sacs was 36.8 mm average; the values for hCG average were 6440 mU/ml and the maximal time of negativization was 52 days. One patient required laparoscopy and salpingostomy, lineal, for inminent rupture 24 hour after methotrexate, the other 14 presented with complete remission. There were no colateral effects. From the 15 patients, in 6 tubal permeability was confirmed by laparoscopy or HSG, being positive in 6 patients, it has not been evaluated, two patients with resolved pregnancy, and other on course (two of them with one salpinx). The proposed management seems to be useful in ectopic pregnancy with success, equivalent to surgical management, and other programs of medical management, with out side effects and with greater easiness of administration. PMID- 10363429 TI - A novel treatment of patients with chronic hepatitis C. AB - OBJECTIVES: Interferon alpha-2b therapy for Chronic Hepatitis C patients has been unsatisfactory. Recombinant Granulocyte Macrophage Colony-Stimulating Factor has been shown to have anti-viral effects in vivo and in vitro via cytokines release. Recently its effects on chronic hepatitis B and possibly chronic hepatitis C were reported. We, decided to conduct a pilot study to evaluate the anti-viral effects of recombinant human GM-CSF mono-therapy in patients with chronic hepatitis C and to assess its side effects. METHODS: A total of 10 patients (male/female: 5/5) (age: 34-60, mean: 45) seen in our center between 2/95 to 2/96 were randomly selected to receive recombinant human Granulocyte Macrophage Colony-Stimulating Factor at 125 ug/m2 subcutaneously daily for two weeks followed by three times weekly for another 8 weeks. Biochemical (ALT) and viral (HCV-RNA) responses were measured prior to treatment and at weeks four and eight. Side effects were recorded. RESULTS: Six out of the ten patients treated had significant viral reduction but none became negative. Eight out the ten patients treated showed biochemical improvement and three out of the eight had normalized liver enzymes. Age, sex, stage of the disease did not influence the response but there seems to be a tendency for patients with higher pre-treatment viral level to respond virally. Side effects are minimal and well-tolerated. CONCLUSION: Recombinant human Granulocyte Macrophage Colony-Stimulating-Factor in the dose used has anti viral effects in the majority of the chronic hepatitis C patients studied. Side effects are minimal and well tolerated. Further study with higher doses and longer duration is needed to prove its clinical efficacy in treating patients with chronic hepatitis C. PMID- 10363430 TI - Liver transplantation in Hawaii: the initial five years. PMID- 10363431 TI - Use of complementary and alternative medicine in Hawaii cancer patients. AB - This research investigated complementary and alternative medicine (CAM) use by Hawaii cancer patients. Thirty-six percent of patients used CAM, most commonly religious/spiritual therapy and herbal treatments. CAM use was linked with younger age, female gender, Catholic religion, and more education. More research is needed to inform decision-making. PMID- 10363432 TI - Colorectal cancer: genetics and screening. AB - Colorectal cancer is a common disease in the Western world. Most, if not all, colorectal cancers develop from previously benign adenomas. There are a number of genetic abnormalities including mutations in oncogenes and tumor suppressor genes which either present as a germline, or acquired defects lead to the development of colorectal cancer. Two well-defined hereditary colorectal cancer syndromes exist, hereditary nonpolyposis colorectal cancer syndrome and familial adenomatous polyposis coli, for which genetic testing is possible and advised. Guidelines for screening for colorectal cancer in average, moderate, and high risk patients are available from the American Cancer Society and were updated in 1997. The American Society of Clinical Oncology has published guidelines for genetic testing in a variety of cancers including colorectal cancer. PMID- 10363433 TI - Coronary artery disease in women: a silent killer. AB - Coronary Artery Disease (CAD) is the leading cause of death and disability among post-menopausal women. Contrary to popular belief, women are at a much greater risk for CAD than for breast cancer. For instance, a 50-year-old female faces a 46 percent risk of CAD and 31 percent risk of CAD mortality. In contrast, her probability of developing and dying from breast cancer is only 10 percent and 3 percent, respectively. In comparison to the other cardiovascular diseases such as mitral valve prolapse, peripartum cardiomyopathy, and eclempsia, CAD is most associated with mortality in women. In fact, one in three women die from CAD in this country. PMID- 10363434 TI - Health care fraud enforcement in 1999. AB - In the current legal and political environment, it is apparent that health care providers will be under more scrutiny for fraud and abuse issues than ever before. Fortunately, some of the areas on which government enforcement personnel will be concentrating are known. This article will review the resources to be devoted to fighting health care fraud and discuss the specific areas to be targeted by enforcement officials. PMID- 10363435 TI - Oklahoma notes decline in Haemophilus influenzae: invasive Haemophilus influenzae disease among children aged < 5 years--Oklahoma, 1990-1997. AB - Haemophilus influenzae (Hi) causes many clinical illnesses such as meningitis, bacteremia, epiglottitis, pneumonia, otitis media, sinusitis and tracheobronchitis. Before the introduction of the Haemophilus influenzae type b (Hib) conjugate vaccine in 1988, Hib caused more than 95 percent of invasive Hi disease in developed countries. Conjugate vaccines were licensed for use in children > or = 15 months of age in 1989 and for use in children > or = 2 months of age in 1990. During 1987-1995, the incidence of invasive Hi disease among children < 5 years of age decreased 96 percent in the United States. This report summarizes the trend in invasive disease caused by Hi among Oklahoma children < 5 years of age. The data represents cases reported to the OSDH as part of the infectious disease surveillance system. Invasive Hi disease has been reportable by law in Oklahoma since 1983. PMID- 10363436 TI - Dr. Kelly West and a brief history of the diabetes epidemic of American Indians. PMID- 10363437 TI - [What is your diagnosis? Aneurysm of the abdominal aorta destroying the lumbar spine]. AB - A 58 year old woman suffering from lumbalgia over 3 years is admitted for bilateral cruralgia. Ten years before, she was treated for cervix carcinoma by hysterectomy, pelvic and lumbo-aortic radiotherapy and chemotherapy. On admission, physical examination reveals L2 level hypoesthesia and abolition of deep tendon reflexes. Plain films and CT scan show a lysis of L3, L4 and L5 vertebral bodies and a cuneiform appearance of vertebral body of L2 due to a large abdominal aortic aneurysm. PMID- 10363438 TI - Healing mechanisms in experimental aneurysms. I. Vascular smooth muscle cells and neointima formation. AB - PURPOSE: The purpose of this work is to better define healing phenomena in this model, in an effort to find strategies to improve long term results of endovascular treatment. METHODS: Lateral wall venous pouch aneurysms were constructed on both carotid arteries in 30 pigs. The aneurysms were packed with collagen sponges per-operatively in 25 animals. Angiography, serial histological studies and immuno-histochemistry tests were used to study healing phenomena and measure neointima formation at various time intervals from 1 day to 9 weeks after surgery. GDC embolization was performed in 5 other pigs for comparison with the collagen sponge model. Explants from the neointima at the neck of aneurysms as well as from the parent artery of 8 pigs were prepared in an attempt to grow and to characterize in vitro cells responsible for healing porcine aneurysms using immunocytochemistry and enzymatic assays. To confirm the hypothesis that an analogy exists between cells involved in aneurysmal healing and neointimal cells found in restenosis, explant outgrowths were scored and compared to explants from intact carotid arteries and carotid arteries subjected to angioplasty in 3 other animals. In addition, to test the value of neointima measurements in quantifying results, 6 dogs were analysed to correlate the thickness of the neointima formed at the neck of aneurysms with angiographic results in animals prone to recurrences. RESULTS: Histopathological findings with collagen sponge packing were similar to the ones following coil embolization. Porcine aneurysms had a strong tendency to heal with a thick neointima primarily composed of vascular smooth muscle cells (VSMCs). Aneurysms in dogs did not heal as well and the neointima at the neck of treated lesions was thin. Cells responsible for healing of experimental porcine aneurysms could be cultured in vitro, and are activated VSMCs. These cells, similar to those harvested following balloon injury, had a higher colony forming capacity and an accelerated explant outgrowth rate as compared to cells derived from the parent artery. CONCLUSION: Animals which heal poorly harbor a thin or deficient neointima at the neck of treated aneurysms. Favorable healing in porcine aneurysms involves VSMCs which form a thick neointima. These VSMCs can be cultured in vitro. They share similar outgrowth characteristics with VSMCs recovered after balloon angioplasty. The collagen sponge model may be useful to harvest cells for in vitro experimentation and in the in vivo evaluation of the local delivery of potential therapeutic molecules thought to improve healing following embolization of aneurysms. PMID- 10363439 TI - [Tubercular lesions of the posterior vertebral arch]. PMID- 10363440 TI - [Contribution of spiral CT scan and MRI in spinal tuberculosis]. AB - Spinal tuberculosis is the most frequent skeletal involvement in tuberculosis. The purpose of this study was to demonstrate the importance of the MRI and the helicoidal CT scan in the diagnosis of spinal tuberculosis. A retrospective study was conducted in 23 patients with spinal tuberculosis. The methods of investigation were helicoidal CT scan in 15 patients and MRI in the other eight patients. In the all cases with helicoidal CT scan, the features of the spinal tuberculosis were seen as an anterior vertebral body destruction with a paraspinal or epidural extension in 12 cases and a sequestrum formation in 5 cases. The analysis with helicoidal CT scan showed a gibbous deformity in 5 cases, a disk space narrowing in 14 cases and evaluated the extension very well. The MR imaging features showed one case in an early stage without features on the plain radiographs, and detected 3 cases of intramedullary lesions. A disruption of the longitudinal posterior vertebral ligament was found in one case and skip lesions at the granulomatous stage in 2 cases. MRI is superior to CT scan even helicoidal CT at the early lesion stage, skip lesions and the ligamentous or medullary lesions. For other lesions (bone, disk, extension), the MRI and CT scan are the same with an advantage for CT scan in osseous lesions. PMID- 10363441 TI - [Efficacy of lumbar chemonucleolysis in the treatment of foraminal and extra foraminal hernias]. AB - A retrospective review of 1,200 nucleolyses performed over a three year period from January 1995 to December 1997 showed 116 discal treatments for foraminal and extra-foraminal localizations. The anatomic localization through the foramen and the intense clinical signs typify these anatomoclinical forms of lumbar hernia. In contrast with complex and disruptive surgical cure, percutaneous nucleolysis offers a precise and rapid method which can be performed in a very short time in outpatients with a success rate of 80%. Most localizations concern L3-L4 and L4 L5 hernias which give the best clinical outcomes. PMID- 10363442 TI - [Evolution of angiographic signs of venous hypertension and clinical signs of intracranial hypertension in intracranial dural arteriovenous fistulas]. AB - Dural arteriovenous fistulas (dAVFs) can cause cerebral venous hypertension (VHT). The most common mechanism is due to the fact that some dAVFs can drain retrogradelly in cortical (better defined as leptomeningeal) veins (directly or after drainage in a dural sinus) causing venous engorgement and consequently an impairment of the cerebral venous drainage. However, more rarely, dAVFs without a cortical venous drainage can also be responsible for VHT probably due to dAVF shunts causing insufficient antegrade cerebral venous drainage. In addition, dAVFs are often associated with stenosis and/or thrombosis of dural sinus(es) which can worsen the VHT. Raised pressure within the superior sagittal sinus causes impeded cerebrospinal reabsorption in the arachnoid villi allowing increased intracranial pressure. The venous engorgement in the cortical veins can cause a venous congestive encephalopathy analogous to the venous congestive myelopathy of the spinal dural AVFs. Clinically VHT can cause not only symptoms related to increased intracranial pressure but also seizures, neurological deficits, impairment of the cognitive functions and dementia. An important aspect is the risk of hemorrhage in dAVFs with a leptomeningeal venous drainage leading to VHT. Although the term VHT sensu strictu should be used if venous pressure measurements are performed, angiographic criteria for VHT such as delayed circulation time, venous engorgement and abnormal visualization of the cerebral veins are well established. The purpose of our study was to evaluate the angiographic signs of VHT in patients with dAVF and to study the course of the VHT and of the clinical signs of increased intracranial pressure before and after dAVF endovascular treatment. A retrospective chart analysis of 22 patients (13 males, 9 females) ranging in age from 20 to 87 years (mean: 53 ys.) with a dAVF associated with angiographic signs of VHT was performed. Ten dAVFs were located on the transverse/sigmoid sinus(es), 6 on the superior sagittal sinus, 3 on the petro-tentorial incisura, 1 on the inferior petrosal sinus, 1 on the anterior ethmoidal region and 1 on the Galen vein region. All dAVFs had a retrograde leptomeningeal venous drainage. Stenosis or thrombosis of the dural AVF sinus was observed in 17 cases and stenosis or thrombosis of another sinus(es) and/or of the jugular vein in 8 cases. In 11 patients, the angiographic signs of VHT were global affecting the entire cerebral venous drainage and, in the other 11 patients, the VHT was focal. The VHT caused clinical symptoms of increased intracranial pressure in 18 patients. Other clinical findings included: bruit (11 cases), seizures (3 cases), vertigo (3 cases), visual deficits (2 cases) and impairment of cognitive functions (4 cases). Three patients presented hemorrhage (one parenchymal hematoma, one hemorrhagic infarction and one subarachnoid hemorrhage). The 4 patients without clinical symptoms of increased intracranial pressure presented only bruit in 2 cases, bruit and vertigo in 1 case, bruit and hemorrhagic infarction in another one. The dAVFs were treated by endovascular therapy (arterial approach: 3 cases, venous approach: 6 cases and both arterial and venous approach: 13 cases). Endovascular sessions ranged from 1 to 7 (mean: 2.8) for each patient. After the endovascular treatment, in 12 patients with complete occlusion of the dAVF, the disappearance of angiographic signs of VHT and clinical cure were observed. In 8 patients with partial occlusion of the dAVF, the disappearance of angiographic signs of VHT and clinical cure were observed in 4 cases (almost complete dAVF occlusion in 2 cases); in the other 4 cases, only reduction the angiographic signs of VHT and clinical improvement were obtained. In all 16 patients who were clinically cured angiographic signs of VHT disappeared despite the persistence of dAVF shunts as observed in 4 cases. (ABSTRACT TRUNCATED) PMID- 10363443 TI - [Guidelines for quality control of cerebral magnetic resonance spectrometry (I). Sequences, spectral quality, control populations. French Network of Clinical Cerebral Spectrometry]. PMID- 10363444 TI - MR imaging of pachydermoperiostosis. AB - A case of pachydermoperiostosis who demonstrated the whole syndrome (pachyderma, periostitis, and cutis verticis gyrata) is presented, and the Magnetic Resonance Imaging (MRI) appearances of the long bone and scalp changes are demonstrated. MRI of the cruris demonstrated fluffy periosteal new bone formation that encroached on the medullary cavity as well as expansion of the diaphysis. Cranial changes included thickening of the diploe associated with diminished signal of the intradiploic fat, and thickening of the scalp with furrowing. PMID- 10363445 TI - Spontaneous chronic spinal epidural hematoma of the lumbar spine. AB - We report an exceptional description of a spontaneous chronic spinal epidural hematoma presenting as lumbar radiculitis. The computed tomographic, magnetic resonance imaging, and intraoperative findings are presented. We discuss anatomical and pathophysiological considerations that could lead to such a condition. We estimate that spontaneous spinal epidural hematomas located in the ventral space are in fact premembranous or posterior longitudinal ligament hematomas. PMID- 10363446 TI - Tolosa Hunt syndrome: a case report. Clinical and magnetic resonance imaging findings. AB - A 36-year-old woman was admitted with a left abducens nerve palsy. MR showed enlargement of the left cavernous sinus. The patient was treated with 80 mg oral methyl prednisolone. Clinical findings improved within a month. Two months later, she was readmitted with left oculomotor and right abducens nerve palsy. MR showed significant increase in the volume of the abnormal area in the left cavernous sinus and a new lesion within the right cavernous sinus. After intravenous gadolinium DTPA, there was enhancement in both cavernous sinuses. Methyl prednisolone therapy was again started. After one month of treatment neurological examination was normal. Follow-up MR findings were similar to previous ones. PMID- 10363447 TI - [Epidemiology, clinical study and pathology of vasospasm]. AB - Symptomatic vasospasms or delayed cerebral ischemia associated with arteriographic evidence of arterial constriction is currently the most important cause of morbidity after acute subarachnoid hemorrhage. Symptomatic vasospasm usually develops between 4 and 12 days after subarachnoid hemorrhage. There is typically a gradual deterioration of the level of consciousness accompanied by focal neurological deficits. 30% of patients who survived aneurysmal SAH develop delayed cerebral ischemia secondary to vasospasm. Vasospasm produces cerebral ischemia and infarction by hemodynamic mechanisms. Vasospasm is an important independent predictor of poor outcome after aneurysmal SAH. Other conditions than aneurysmal subarachnoid hemorrhage such as trauma, tumors, unruptured aneurysms, meningitis and ruptured AVM may be associated with vasospasm. PMID- 10363449 TI - [Transcranial Doppler and cerebral vasospasm]. AB - Increase in transcranial Doppler ultrasound flow velocities in the major basal arteries correlates with symptomatic vasospasm. Transcranial Doppler examinations are performed using a pulsed Doppler Probe via the trans temporal approach. Transcranial colour-coded real time sonography can be useful and help to identify the cerebral arteries. Maximum flow velocities of > 200 cm/sec are associated with cerebral ischemia and infarction. A maximum rise in Doppler velocity of more than 50 cm/sec/24 h is correlated with poor outcome. Using a diagnostic cutoff of 130 cm/sec a 87% positive predictive value can be obtained using TCD in the middle cerebral artery. Numerous factors affect Doppler flow velocities and may lead to erroneous conclusions about the presence or absence of vasospasm. Flow velocity is directly related to cerebral blood flow. Intracranial pressure, blood pressure and volume, hematocrite and subarachnoid hemorrhage affect Doppler flow velocities. False-negative examinations of vasospasm using TCD are associated with distal vasospasm, severe spasm of the carotid siphon, chronic high blood pressure and increased intracranial pressure. PMID- 10363448 TI - [Imaging of vasospasm]. AB - Neuroimaging is of paramount importance in the evaluation and management of cerebral vasospasm. Arteriographic demonstration of concentric arterial constriction between 4 and 12 days after subarachnoid hemorrhage is the definition of vasospasm. Assessment of angiographic vasospasm is subjective. Vasospasm must be differentiated from hypoplasia, atherosclerosis and other non specific arteriographic conditions. The development of vasospasm is directly correlated with the presence of thick blood clots in the basal subarachnoid cisterns which can be detected by an early computed tomography scan. MR imaging and evaluation of cerebral blood flow are useful in the early identification of cerebral ischemia. PMID- 10363450 TI - [Pathophysiology and principles of treatment of vasospasm]. AB - The exact mechanism of vasospasm is still unknown. The etiology of cerebral vasospasm is subarachnoid blood clot. Vasospasm is a multifactorial process. Oxyhemoglobin is released by erythrocyte lysis and exert several effects on the endothelium that could lead to vasoconstriction. The production of free radical and superoxide anion radical secondary to hemoglobin degradation stimulates the release of vasoconstricting products. Arterial vasoconstriction secondary to smooth muscle contraction could be related to increase in protein kinase C. Narrowing of cerebral vessels produces cerebral ischemia by hemodynamic mechanisms. Direct hypothalamic insults may be associated. Clot removal and clot lysis have been proposed to prevent vasospasm. Pharmacological treatments are targeted to the vasospasm itself (nicardipine, AT 877) or the prevention of delayed ischemic events (nimodipine, tirilazad). General measures such as the "triple H therapy" (hemodilution, hypertension, hypervolemia) are widely used in the prevention and/or treatment of cerebral vasospasm. PMID- 10363451 TI - [Endovascular treatment of vasospasm]. AB - Cerebral vasospasm remains a leading cause of heightened morbidity and mortality rates following aneurysmal subarachnoid hemorrhage despite the apparent benefit of recent medical therapeutics. Successful resolution of medically refractory angiographically demonstrated vasospasm with concomitant reversal of delayed neurological deficit has been observed after balloon angioplasty. Subsequent reports confirmed these encouraging results but also emphasized the limitations of the technique and the risks of complications. Intraarterial papaverine infusion has been performed for the treatment of diffuse cerebral vasospasm with controversial results and has also been combined with angioplasty either to facilitate balloon navigation or to treat arteries inaccessible to balloon catheterization. All these different endovascular approaches explain the confusion existing about the indications, timing and efficacy of the endovascular treatments. This article reviews several clinical and experimental studies dealing with these questions. PMID- 10363452 TI - [Management of vasospasm from subarachnoid hemorrhage. Attitude of French centers. French Society of Neuroradiology]. PMID- 10363453 TI - [Practical aspects of realization of a functional MRI]. AB - Functional MRI (fMRI) allows cognitive processes to be studied on an individual basis. fMRI is at the crossing of many fields, such as physiology, MRI physics, cognitive neurosciences, data analysis,.... Hence, it requires the collaboration of inter-disciplinary teams, in which neuroradiologists should play an important role. This article provides an overview of the general principles necessary to conduct fMRI studies on a practical matter (choice of magnetic field, patient set up and preparation, paradigm design, data acquisition and analysis). PMID- 10363455 TI - [Brain plasticity during development: physiological bases and functional MRI approach]. AB - Brain functional MRI (fMRI) is a new tool for the study of the development of cognitive functions in healthy children ("natural plasticity"), as well as for the assessment of functional reorganization following brain lesions. However, methodological difficulties related to the pediatric population (movements, cooperation), along with unsolved issues about the influence of physiological parameters of the immature brain on fMRI results, explain the limited number of published studies. Normal brain maturation is characterized by a transient phase of synaptic redundancy followed by selective synaptic regression until adulthood, that forms the neurobiological correlates of both learning and individual variability of cortical anatomy and functional organization, and of the large potential for post-lesional plasticity in children. fMRI in school-age children demonstrated activation patterns comparable to adults during motor, language, and working memory tasks. In neonates and infants, fMRI showed significant differences of visual cortex activation. Post-lesional plasticity is more pronounced in younger children. In motor cortex, activation of ipsilateral hemisphere may be seen in cases of rolandic lesions. Interhemispheric shift of language networks occurs mostly in cases of destructive or large brain lesions, or in cases of early refractory epilepsy. PMID- 10363454 TI - [Influence of individual strategies on brain activation patterns during cognitive tasks]. AB - Functional magnetic resonance imaging (fMRI) at 1.5 T was used to investigate the influence of cognitive strategies on cortical activation during mental calculation. Twenty-nine right-handed subjects performed a serial subtraction of prime numbers. Data were analyzed taking into account whether the spontaneous strategy of subjects was verbal (n = 15) or visual (n = 14). Even thought a common corpus of brain areas was activated during this mental calculation task, i.e. the dorsolateral prefrontal, premotor and parietal cortices, and Broca's area, differences appeared between the two groups of subjects. In subjects using a verbal strategy, the main activation was located in the whole left dorsolateral frontal cortex with a little activation of the inferior parietal cortices. In subjects using a visual strategy, a bilateral activation in the prefrontal cortex and a high activation in the left inferior parietal cortex were observed. These results demonstrate that numbers are processed through a distributed network of cortical areas, the lateralization of which is clearly influenced by subject strategy. Taken together this data reveals a functional interaction between the left inferior parietal cortex and the right prefrontal cortex in the visuo spatial sketchpad for number processing. This network could be involved in sustained selective attention to mental numerical images generated in the left inferior parietal cortex. PMID- 10363456 TI - [Functional MRI: brain plasticity, brain disease and functional recovery]. AB - Brain plasticity may be defined as long-term alteration in behavior related activity of distributed neural systems. Functional imaging, and particularly functional MRI, allows to investigate the mechanisms underlying brain plasticity. Two aspects may be distinguished: one is learning in healthy subjects; the other is functional reorganization following or associated with acute or chronic brain injury. Because functional MRI is totally non invasive, it appears well suited to follow such reorganization over time. This theme will be developed in the following text. PMID- 10363457 TI - [Importance and limitations of the validation of functional MRI of motor function and language using preoperative cortical stimulation]. AB - This chapter describes and discusses the value of the localization of functional areas obtained from functional MRI in brain tumor cases. Correlation method is cortical brain mapping by intraoperative stimulation. The experience reported here is focused on the study of motricity and language. METHODS: Twenty two patients with tumors of the rolandic region (n = 16) or in the temporal lobe (n = 6) underwent functional MR mapping and subsequently cortical mapping before tumor resection. The tasks chosen were a flexion and extension of the fingers or a naming task. We used 3D reconstructed images of the surface of the brain to assess intra and post operatively the functional MRI and stimulation data. RESULTS: For the motor correlation, in each case, the results of direct cortical mapping matched those obtained with functional MRI, both positively and negatively, although the extent of the functional activations was larger than the area required to elicit the corresponding movement during intraoperative brain mapping. For the language correlation and for the task chosen, only the results of the precentral areas matched those of functional MRI. CONCLUSIONS: Functional MRI can be used preoperatively to assess motor functional area in patients with rolandic tumors. More studies are needed to validate intraoperatively the language areas and the real extent of functional MRI activations. Finally, the observed discrepancy between functional MRI and cortical stimulation is likely due to the rather profound differences between both techniques, in terms of neurophysiology, practical applications and statistical analysis. PMID- 10363458 TI - APhA drug treatment protocols: self-care of self-limited pain. PMID- 10363459 TI - Visits to office-based physicians in the United States for medication-related morbidity. AB - OBJECTIVE: To examine the prevalence, nature, demographics, and resource use associated with visits to office-based physicians in the United States during 1995 for medication-related morbidity. DESIGN: A nationwide cross-sectional survey of ambulatory care visits to physician offices, based on data from the National Center for Health Statistics' 1995 National Ambulatory Medical Care Survey. SETTING: Physician office-based settings in the United States. PATIENTS: Patients visiting office-based physicians for principal diagnoses of adverse effect of medications (ICD-9-CM E-code 930.00-947.9). MAIN OUTCOME MEASURES: Weighted measures of prevalence, nature, demographics, and resource use associated with visits related to adverse effects of medications. RESULTS: An estimated 2.01 million (95% confidence interval, 1.69 to 2.34 million) visits for medication-related morbidity were made to office-based physicians in the United States during 1995, representing an annual rate of 7.70 visits per 1,000 persons. Medication-related visit rates were greater in women, in patients between 65 and 74 years of age, and in the Midwest. The most frequently cited reasons for medication-related visits were skin rash, nausea, and shortness of breath. The therapeutic agents responsible for medication-related visits were most often hormone and synthetic substitutes (13.32%), antibiotics (11.55%), and cardiovascular drugs (9.30%). Medication-related visits most often involved diagnostic services and medication therapy. The majority included instructions for a scheduled follow-up, and fewer than 1% resulted in hospital admission. CONCLUSION: Medication-related ambulatory care utilization can pose a significant burden on health care resources unless specific strategies are initiated to control medication-related problems. The provision of pharmaceutical care can play an important role in reducing medication-related problems and associated health care costs. PMID- 10363460 TI - Evaluating the use and quality of pharmacy drive-up services. AB - OBJECTIVE: To evaluate the use and quality of pharmacy drive-up services; specifically, to assess patients' and pharmacists' views of the drive-up and determine the implications for assuring or improving the quality of the services provided. DESIGN: Questionnaires developed for patients and pharmacists addressed aspects of pharmacy drive-up services. The patient questionnaire asked about prescription information and drive-up use, satisfaction with pharmacy services, importance of patronage factors, comfort in using the drive-up, and comparison of drive-up and in-store visits. The pharmacist questionnaire was similar in content, but also contained open-ended questions pertaining to the provision of pharmaceutical care at the drive-up pharmacy. SETTING: Six pharmacies in Central Iowa. PARTICIPANTS: A sample of patients who had received at least one prescription from a participating pharmacy in the previous 30 days; pharmacists from participating pharmacies. MAIN OUTCOME MEASURES: Responses from users and nonusers of the pharmacy drive-up. RESULTS: Both users and nonusers of the drive up indicated that pharmacist-linked services--those that entail direct pharmacist patient contact and are closely associated with the ideals of pharmaceutical care -may be provided better in-store. The pharmacists also indicated that these services were provided better in-store and mentioned other ways in which drive-up services may detract from patient care. CONCLUSION: As pharmacist-linked services become more important in health care, and as drive-up pharmacy services increase in popularity, the compatibility of drive-up service with a greater emphasis by pharmacists on patient care will increasingly become an issue. To allay potential concerns and improve the quality of service, pharmacists need to take additional steps to assess drive-up patients' level of familiarity with their medications, require patients to come into the pharmacy periodically, and, where necessary, provide alternative services at the drive-up. PMID- 10363461 TI - Effect of a change in third party reimbursement rate on prescription gross margin. AB - OBJECTIVE: To measure the effect of a change in an insurance company's reimbursement formula on prescription department gross margins for all prescriptions and subgroups of prescriptions. DESIGN: Retrospective descriptive analysis. SETTING: Wisconsin. PARTICIPANTS: Two units of a chain pharmacy. INTERVENTION: Reimbursement changed from usual and customary price to average wholesale price less 10% plus a $2.00 dispensing fee for single-source products, and maximum allowable cost plus a $2.00 dispensing fee for multisource products. MAIN OUTCOME MEASURE: Gross margins for prescriptions dispensed in the month before and after the reimbursement change. RESULTS: The average estimated gross margin decreased 26.9% after the change in reimbursement, and the effect on the average gross margin for generic prescriptions was nearly twice that of the effect on the average gross margin for brand name prescriptions. The effect of the reimbursement change on different therapeutic classes ranged from an increase of 0.7% in the cardiovascular class to a decrease of 68.2% in the eyes, ears, nose, and throat class. The effect of the reimbursement change was greater for low-cost prescriptions than for high-cost prescriptions. CONCLUSION: The large effect of the reimbursement change, combined with continued growth in third party prescriptions, raises concerns about whether pharmacies can accept third party contracts with low reimbursement rates and still maintain current profitability and service levels. PMID- 10363462 TI - Career commitment: a mediator of the effects of job stress on pharmacists' work related attitudes. AB - OBJECTIVE: The hypothesis for this study was that career commitment mediates the effects of job stress on several work-related attitudes of pharmacists. The effects of job stress, career commitment, met expectations, job satisfaction, and organizational commitment on job turnover intention were also investigated. DESIGN: Cross-sectional mail survey. SETTING: Nationwide sample of licensed pharmacy practitioners in the United States. PARTICIPANTS: 1,088 respondents (full- or part-time chain, independent, or hospital pharmacists) to a previous study. MAIN OUTCOME MEASURES: Rating scales measured each of the study variables. Demographic information was also collected. Data analyses included descriptive statistics, confirmatory factor analysis, and structural equation modeling with latent variables. INTERVENTIONS: None. RESULTS: Responses were received from 653 pharmacists out of 921 contacted (71%). The effects of job stress on job turnover intention were mediated through career commitment, met expectations, organizational commitment, and job satisfaction. Career commitment positively affected met expectations (beta = 0.35), and met expectations positively affected organizational commitment (beta = 0.66) and job satisfaction (beta = 0.78). An increase in the mean level of job satisfaction and organizational commitment decreased the likelihood of job turnover intention. Independent pharmacy owners tended to have the most positive attitudes toward work. CONCLUSION: Strategies should be developed to increase the career commitment of pharmacists. Increased commitment can reduce the negative effects of job stress and improve work-related attitudes. PMID- 10363463 TI - Pharmaceutical care certificate program: assessment of pharmacists' implementation into practice. AB - OBJECTIVE: To measure the effects of a Pharmaceutical Care Certificate Program (PCCP) in community pharmacists. DESIGN: This study compared the effects of the PCCP over time using a repeated measures design. SETTING: Retail, independent, and managed care pharmacy. PARTICIPANTS: 36 pharmacists who participated in the PCCP. INTERVENTIONS: PCCP, which was developed to train pharmacists in (1) practice re-engineering, (2) components of pharmaceutical care, and (3) drug therapy management of disease states. MAIN OUTCOME MEASURES: (1) pharmacist job functions, (2) pharmaceutical care job functions, (3) pharmaceutical care components, (4) pharmacist-perceived barriers to providing pharmaceutical care, and (5) proposed solutions for overcoming barriers. RESULTS: When comparing pharmacists at baseline and after 1 year, pharmacists after 1 year felt significantly better prepared to perform all pharmaceutical care components. CONCLUSION: Although the PCCP was successful in preparing pharmacists to perform the pharmaceutical care components covered in the program, time seems to be one of the major barriers for their actual implementation into practice. One proposed solution is the development of partnerships between pharmacists and schools of pharmacy. PMID- 10363464 TI - Establishing community pharmacy-based anticoagulation education and monitoring programs. AB - OBJECTIVE: To determine the process for establishing community pharmacy-based anticoagulation education and monitoring programs using fingerstick capillary whole blood testing. DESIGN: Pilot community-based intervention study using convenience sample of patients. SETTING: Three community pharmacies with pre established health education centers and laboratories certified for moderate complexity. PARTICIPANTS: 26 patients were referred to the clinics by 2 primary care physicians for each pharmacy, most with a diagnosis of atrial fibrillation. INTERVENTION: Patient assessment, including adherence to prescribed regimens; changes in medication use, including prescription and nonprescription medications, vitamins, health foods, and nutrition supplements; changes in diet and ethanol consumption; assessment of adverse experiences; and needed changes in warfarin dosage. MAIN OUTCOME MEASURES: Percentage of international normalized ratio (INR) values within therapeutic range, major bleeding events, and thrombotic events. RESULTS: A total of 21 patient charts were available for analysis. More than 80% of patients had INR values within their targeted range (+/- 0.2) 60% or more of the time, comparable with values reported for anticoagulation clinics. Of the 235 INR values obtained during the study, 75% were within the individualized targeted therapeutic range (e.g., 2 to 3 +/- 0.2). One patient experienced a major bleeding event related to an underlying cancer. None of the patients experienced a thrombotic event. CONCLUSION: Community pharmacies can effectively implement an anticoagulation education and monitoring program. PMID- 10363465 TI - Stability of amlodipine besylate in two liquid dosage forms. AB - OBJECTIVE: To determine the stability of amlodipine besylate in two liquid dosage forms under refrigeration and at room temperature. DESIGN: Commercially available amlodipine tablets (Norvasc-Pfizer) were used to prepare two suspensions: one in extemporaneously prepared 1% methylcellulose in syrup (1:1), and another in equal volumes of commercially available OraPlus/OraSweet. Each suspension containing amlodipine 1 mg/mL was stored in 10 plastic prescription bottles; 5 were stored at 4 degrees C and 5 at 25 degrees C. Samples were collected immediately after preparation (day 0) and on days 7, 14, 28, 42, 56, 70, and 91. Amlodipine concentration was measured by stability-indicating HPLC method (n = 15). SETTING: Research laboratory at Children's Hospital. MAIN OUTCOME MEASURES: Physical and chemical stability (> 90% of the initial concentration) of amlodipine in the two extemporaneously prepared suspensions during storage in plastic prescription bottles at 4 degrees C and 25 degrees C. RESULTS: Observed mean concentrations exceeded 90% of the initial concentrations in both suspensions for 91 days at 4 degrees C and 56 days at 25 degrees C. No noticeable change in physical appearance or odor was observed; pH changed slightly in the methylcellulose containing formulation stored at 25 degrees C. CONCLUSION: Amlodipine was stable in two suspensions when stored in plastic prescription bottles for 91 days at 4 degrees C or 56 days at 25 degrees C. These formulations may be considered for pediatric or elderly patients who are unable to swallow tablets. The liquid dosage form would also permit accurate administration of amlodipine doses to infants and young children based on their body weight. PMID- 10363466 TI - Eating disorders: current concepts. AB - OBJECTIVE: To review for the community pharmacist common eating disorders and suggest strategies for pharmacist-directed patient education. DATA SOURCES: Current literature. DATA SYNTHESIS: Women are much more likely than men to develop eating disorders, and Western culture's emphasis on thinness contributes to the prevalence of eating disorders in the United States. Regardless of the type, all eating disorders are rooted in emotions, often traced to problems during adolescence. Anorexia nervosa patients continually set and strive to obtain lower goal weights to the point that their general well-being is compromised. Bulimia nervosa is characterized by excessive eating followed by purging that is an apparent attempt to relieve the tension and guilt associated with the initial overeating. Rather than directly dealing with tension and anger, a patient suffering from binge eating overindulges in food. Most overeaters are aware that excess weight is detrimental to their health, but they cannot control their overeating behavior. CONCLUSION: In treating eating disorders, it is important to address patients' emotional and psychological needs as well as physical symptoms. Patients often need encouragement to seek and continue treatment, and pharmacists are in an ideal position to provide that support. PMID- 10363467 TI - Applying managed care performance measures in community pharmacy-based outcomes research. AB - OBJECTIVES: (1) To introduce the National Committee on Quality Assurance's (NCQA's) Health Employer Data and Information Set (HEDIS), a set of managed care performance measures, as a tool for conducting outcomes research in the community pharmacy; (2) To demonstrate the practical application of this tool by describing three outcomes research projects conducted in an independent community pharmacy. DESIGN: Review of three outcomes research projects based on HEDIS: (1) Increasing ACEI Use in Patients with Heart Failure, (2) Cardiovascular Telepharmacy Project, and (3) Pediatric Antibiotic Callback Program. SETTING: Independent community pharmacy. RESULTS: The "Effectiveness of Care" section of HEDIS was a practical tool for designing outcomes research projects in the community setting. Improved clinical and/or service outcomes were observed in two of the projects. The challenges faced in these projects were due to the methods of implementation, rather than to the HEDIS tool itself. These projects led to a number of "lessons learned" in the conduct of outcomes research in the community pharmacy setting. CONCLUSION: Community pharmacists can use NCQA's HEDIS as a tool to help gain acceptance of and reimbursement for pharmaceutical services through the formulation of outcomes projects that are of interest to managed care organizations (MCOs) seeking accreditation status. As pharmacists report their successes to MCOs, they may be better equipped to develop reimbursement strategies for services from MCOs through their demonstration of shared goals. PMID- 10363468 TI - Development of pharmaceutical care in The Netherlands: pharmacy's contemporary focus on the patient. AB - OBJECTIVE: To describe how developments in the pharmacy profession in The Netherlands converged into the current movement toward pharmaceutical care. SETTING: Dutch community pharmacy. DESCRIPTION: Literature was reviewed for key elements of pharmacists' professional development over the last 40 years--the pharmacist-physician relationship, the pharmacist-patient relationship, the education of the pharmacist, provision of information to patients, medication surveillance, clinical pharmacy, and social pharmacy. Consideration was given to how, when and if these elements interacted and contributed to the movement toward pharmaceutical care. RESULTS: During the early years of the 20th century the professional role of the pharmacist, based on preparing medications, declined because of the increased industrial production of drugs. In The Netherlands, a number of developments, starting around 1995, led to a "reprofessionalization" movement in pharmacy, characterized by pharmacists' increased awareness of social and ethical responsibilities with respect to drugs and patients. These developments included an improved relationship between pharmacists and physicians, the implementation of clinical pharmacy and medical surveillance in daily community pharmacy practice in the 1970s and 1980s, and the increased awareness of the rights of patients to quality drug information and counseling in the 1980s and 1990s. By the end of 1980 these trends had coalesced into a professional movement supporting the need for a pharmaceutical care model of practice. CONCLUSION: Dutch pharmacy is gradually implementing pharmaceutical care in daily community practice. However, a proactive attitude, not only from the "front runners," but from all pharmacists, is desirable if pharmaceutical care is to be incorporated into routine community practice. PMID- 10363469 TI - Designing solutions for securing patient privacy--meeting the demands of health care in the 21st century. AB - OBJECTIVES: To define the issues surrounding patient privacy, examine the political context in which debate is taking place, and present a novel technology model for addressing privacy, confidentiality, and security in 21st century health care. SUMMARY: The discussion of privacy addresses one of the basic issues in health care today--the tension between the needs of the individual patient for privacy and confidentiality and the needs of society to effectively manage health care practices and control health care costs. Patient concerns for privacy, confidentiality, and security are legitimate, and can usually be reduced to issues that potentially affect an individual's employment, ability to get and maintain health coverage, and have control over his or her records and care. These concerns, combined with several precipitating events, are forcing the issue of privacy into the political arena, where new health policy decisions will be made. The debate must be framed within a principle-centered approach that focuses on boundaries, security, consumer control, accountability, and public responsibility. A global, distributed electronic health record management model that provides location-independent, secured, authenticated access to relevant patient care records by qualified health care professionals on a need-to-know basis provides solutions. Information asset considerations should be designed to equitably represent the ownership needs of corporate entities, society, and the individual. CONCLUSION: A secure electronic health record structure that systematically ensures a high level of accountability combined with thoughtful dialogue among key stakeholders in the public policy development process can offer the privacy outcomes we seek. PMID- 10363470 TI - Establishing a lipid management service in the pharmacy. PMID- 10363471 TI - Etanercept: a new option in the management of RA. PMID- 10363472 TI - Ginkgo biloba for dementia: a reasonable alternative? PMID- 10363473 TI - Vaccine side effects: separating mirage from reality. PMID- 10363474 TI - Survey of primary care training in Louisiana. LSMS Committee on Medical Education. PMID- 10363475 TI - Preparing Medicare for the 21st century. AB - On July 1, 1966, the federal government offered assistance in paying for healthcare to almost all Americans aged 65 or older. The Medicare program has been an incredibly successful program ever since then, but is now facing severe financial and other challenges that require serious reforms. Healthcare delivery systems and technology have undergone tremendous change during the past three decades, and it is time to modernize the Medicare program to reflect these changes and prepare the program for the next century. PMID- 10363476 TI - ECG of the month. Stop recycling! AV junctional reentrant tachycardia. PMID- 10363477 TI - Elective surgery and the HIV-positive patient: medical, legal, and ethical issues. AB - The ethical and legal issues surrounding the healthcare provider's obligation to provide care for patients have been a topic of debate since the beginning of modern medicine. The human immunodeficiency virus (HIV)-positive patient requesting cosmetic or elective surgery provides yet another situation in which the physician's ethical and legal responsibilities for the patient become a topic of debate. The risks involved to the physician and patient are first discussed, and then current ethical theory and legal decisions are reviewed. Finally, some conclusions are attempted from the varied opinions in the literature surrounding this controversial topic. PMID- 10363479 TI - The journal 150 years ago. May 1849. PMID- 10363478 TI - Radiology case of the month. Nail puncture wound to the foot. Mycobacterium chelonei osteomyelitis. PMID- 10363480 TI - Current treatments of atrial fibrillation. AB - Atrial fibrillation is the most common arrhythmia encountered in medical practice and is responsible for significant morbidity and mortality and consumes much of healthcare's finances. Controlling the patient's ventricular rate becomes the first priority, and digoxin, calcium channel blockers, and beta blockers all come under consideration for the individual patient. Evaluation and treatment of contributing factors such as thyrotoxicosis, acute infarct, or ethanol consumption must also always be addressed. Consideration can then be given to whether a patient should be anticoagulated and cardioverted and to which medications or procedures are appropriate. The special situation of atrial fibrillation after cardiac surgery is also a frequent challenge, with various treatment options available. PMID- 10363481 TI - Coronary interventions approaching the year 2000. AB - Interventional cardiology is only 22 years old, but its successes at the end of this century stand heavily on the shoulders of innovative pioneers who labored earlier in the century. Balloon angioplasty gradually developed and eventually achieved great success in treating America's greatest health risk, coronary artery disease, through the 1980s. Both mortality benefit and symptom improvement have been demonstrated for coronary angioplasty, making it one of the most frequently performed procedures in the world today. In an effort to overcome acute complications and late restenosis, atherectomy devices and stents became useful tools over the past decade. As the generation of baby boomers begins to swell the ranks of the middle aged and elderly in the early 21st century, it is with great hope that molecular biology and the continued development of the technology of interventional cardiology will allow even greater successes in decreasing death and disability from ischemic heart disease. PMID- 10363482 TI - Advances in the pathogenesis and treatment of acute coronary syndromes. AB - The clinical entities of unstable angina, non-Q wave myocardial infarction, and Q wave myocardial infarction share the same pathogenesis and, because of this, are linked under the heading of acute coronary syndromes. Prompt reperfusion in the early phase of acute ST segment elevation myocardial infarction, with thrombolysis or percutaneous transluminal coronary angioplasty, now has an established place in the treatment of this condition. However, thrombolysis has been disappointing and may be harmful in the treatment of unstable angina and non Q wave myocardial infarction. While traditional therapy with morphine, oxygen, nitrates, aspirin, heparin, and beta blockers may be indicated in the treatment of all types of acute coronary syndromes, recent studies have led to advances in the treatment of unstable angina/non-Q wave myocardial infarction patients. In these patients, enoxaparin (a low molecular weight heparin) and the platelet glycoprotein IIb/IIIa receptor antagonists may be particularly effective. PMID- 10363483 TI - Update in cardiac surgery. AB - Cardiac surgery continues to evolve in response to advances in technology, patient demands, and healthcare reimbursement. The development of minimally invasive operative techniques and laser technology has enhanced the therapeutic armamentarium available to the cardiac surgeon. Advances in biomaterial development will see the artificial heart in clinical trials by the millennium. PMID- 10363484 TI - Ventricular tachycardia and sudden cardiac death. AB - As we approach the new millennium, treatment of survivors of cardiac arrest and prevention of sudden cardiac death (SCD) are the two most important problems confronting contemporary cardiology practice. Sudden cardiac death occurs as a result of ventricular tachycardia (VT) degenerating into ventricular fibrillation (VF). Several major arrhythmia treatment trials completed during the last decade have significantly changed the way we treat patients with ventricular arrhythmias. In patients with sustained VT and aborted SCD, only treatment with implantable cardioverter defibrillator (ICD) has been shown to significantly increase survival. Amiodarone and sotalol, though very useful in the treatment of VT and VF, do not improve survival as significantly as ICD therapy. Use of Class I antiarrhythmics may adversely affect survival. Primary prevention of SCD in patients with a recent myocardial infarction (MI) and in patients with cardiomyopathy and congestive heart failure (CHF) is limited by our inability to accurately identify patients at risk of SCD. Among the many tests available to identify patients at risk of SCD, decreased left ventricular ejection fraction (LVEF) and presence of non-sustained VT appear to be most useful. To date, only beta adrenoceptor blockers have been shown to improve survival in post-MI patients as well as in patients with cardiomyopathy and CHF. Use of amiodarone is controversial in these patients. Treatment with ICD of post-MI patients with decreased LVEF and inducible sustained VT at electrophysiology study improves survival. PMID- 10363485 TI - Empirical study of disability, employment policy, and the ADA. PMID- 10363486 TI - [In memory of Dr. Guillermo Dierssen]. PMID- 10363487 TI - [Intranuclear inclusions in the syndromes by dynamic CAG mutations: what is its pathogenic role?]. PMID- 10363488 TI - [The eye-lid reflex in hemifacial spasms. Study of 57 patients]. AB - FUNDAMENTALS: [corrected] The blink reflex is the neurophysiological method for the study of hemifacial spasm. It is a polysynaptic reflex with a primary component or R1, which is of pontine origin, and a secondary component or R2 probably originating on the pons and lateral bulb. OBJECTIVE: To study hemifacial spasm, its clinical, and demographic characteristics and the possible neurophysiological (blink reflex) differences between symptomatic and idiopathic cases. METHODS: The study population included 57 patients (35 women and 22 men) with no axonal damage on the seventh cranial nerve; the control group included 57 healthy people paired up according to age and sex. Patients underwent a CT scan or MRI. Blink reflex was studied on the healthy and damaged sides of the patient group and left side was studied in the control group. After hyperventilation the study was repeated on the affected patient group only. RESULTS: The patients' average age was 53.4 years and the average duration of the disease was 3.7 years. Thirty-four patients had proven structural anomalies and the left side was affected in the same number. When comparing the results between the damaged side, the normal one and the control side it was only significant for the R1 component (p < 0.00000). No differences were found between the symptomatic and the idiopathic cases before or after hyperventilation, that lengthened R1, R2 and R2c latency periods (p < 0.00000; 0.0007 and 0.00000, respectively). CONCLUSIONS: Blink reflex does not enable the establishment of differences between the idiopathic and the symptomatic cases. The rise of the latency periods after hyperventilation signals the beginning of hyperexcitability of the secondary facial motor nucleus due to a peripheral lesion, to loss of supranuclear inhibitory control or both. PMID- 10363489 TI - [Evolution and adverse effects in patients with remittent-recurrent multiple sclerosis treated with interferon beta 1b. The influence of patient's weight and height]. AB - BACKGROUND: The aim of the present study was to determine whether the adverse effects (AE) of interferon beta 1b (IFNb-1b) are related to patient weight, height and body surface area. Moreover, whether the basal incapacity and/or some of these AE may be related to the evolutive prognosis were also studied. METHODS: Twenty-nine females and 14 males with remittent-recurrent multiple sclerosis treated with IFNb-1b were studied. The clinical data and the AE, were compared with the number of outbreaks and the progression of incapacity over the first year of treatment in patients with high weight, height and body surface area with respect to those under the mean. RESULTS: Although the presence of fever was similar in the two groups during the first month (8/19 in the high weight patients and 11/24 in the low weight group), in the following months, overall, the low weight patients continued to present fever. Fever during the first month was associated with a lower number of outbreaks at one year of evolution (0.04 +/ 0.1, n = 19 vs 0.30 +/- 0.5, n = 24; p = 0.04). Furthermore, the patients with lesser incapacity presented a better evolution than those with more incapacity, particularly at 2 years (0.21 +/- 1.3 vs 1.36 +/- 0.9; p = 0.05). CONCLUSIONS: During the first month of treatment fever is very frequent in all the patients regardless of weight and fever was correlated with a lower number of outbreaks at one year of evolution. From the third month fever was less frequent in the lower weight patients group. PMID- 10363490 TI - The PDQ-39 Spanish version: reliability and correlation with the short-form health survey (SF-36). AB - BACKGROUND AND METHODS: The Parkinson's Disease Questionnaire (PDQ-39) was the first specific instrument for evaluation of the "health-related quality of life" (QoL) in Parkinson's disease patients. The PDQ-39 has been subjected to adaptation to Spanish language and culture (PDQ-39 Spanish version, PDQ-39SV) and this version has been validated in aspects of internal consistency and construct validity. The present study assess the test-retest reliability and the convergent validity of the PDQ-39SV with a generic QoL instrument (SF-36). RESULTS: Most of the PDQ-39 dimensions showed an adequate consistency-Cronbach's alpha > 0.7 for six dimensions. As a whole, test-retest reliability resulted satisfactory. Two dimensions-activities of daily living and emotional well-being- showed a low grade significant difference (paired Student t-test, p < 0.05) due to improvement in the second survey (at 10 to 14 days from the first one) perhaps related to adjustments of the treatment at the first visit. A strong association (Spearman r, p < 0.001), indicative of convergent validity, was obtained for the PDQ-39 dimensions and the relevant SF-36 scales, as well as for the physical and mental component summary scores of the SF-36. CONCLUSIONS: Taking into account these results and previous studies, it is concluded that the PDQ-39 SV is a reliable measure that has construct validity. PMID- 10363491 TI - [Quality of life and Parkinson's disease]. PMID- 10363492 TI - [Genetic bases in Huntington disease]. AB - Huntington disease is a neurodegenerative syndrome of late appearance in 80% of the cases. Its clinical course shows motor and cognitive disorders which lead to total incapacity of the individual 10 to 15 years after the appearance of the first syndromes. From a genetic point of view, this disease is of autosomic dominant inheritance with complete penetration. The gene (IT15) is localized in the short arm of chromosome 4 in the telemaric region 4p16.3 and it is known that the mutation is produced by an increase in the number of trinucleotides CAG (glutamine) localized in the 5' end of the gene, in the first exon. In the general population these are repeated in a number of less than 30 repetitions and in the disease population in a number greater than 36 and, in some cases, even greater than 100 repetitions. These sequences are unstable from one generation to another and this fact may explain the phenomenon of genetic anticipation shown in this disease since there is an inversely proportional correlation between the number of repetitions and the age of appearance of the first symptoms. The gene codifies for a protein known as "huntingtina" which is expressed not only in brain but also in different tissues. Although the function remains unknown, it is known that it is bound to other proteins related to the transmission of neuron signals and to the regulation of neuron death (apoptosis) among others. The defect is produced by a gain in function closely related to the number of repetitions. At present, presymptomatic and prenatal diagnosis of the disease may be achieved since the reliability of the studies is practically 100%. Nonetheless, the demand is lower than expected and this may be due to psychologic, social and legal problems, together with the lack of adequate infrastructure for completely guaranteeing the performance of these studies. PMID- 10363493 TI - [Treatment of Alzheimer's disease: acetylcholinesterase inhibitors]. AB - AIM: The cognitive deficiency of Alzheimer's disease is attributed to a dysfunction in the cerebral cholinergic systems. Current drug treatments are directed at stimulating cholinergic transmission. The aim of this study was to evaluate the latest cholinergic drugs available an those about to appear in the market. METHODS: A review of the most recent studies published regarding the physiopathology of Alzheimer disease and the results following treatment with donepezil, rivastigmine and metriphonate was carried out. RESULTS: Donepezil is a specific, reversible acetylcholinesterase inhibitor of close to 100% absorption and a half-life of 70 h, achieving stable concentrations at approximately 3 weeks. Patients treated with a single daily dosis of 5 or 10 mg improve in the ADAS-Cog scale. The medication is initiated with a dosis of 5 mg/day. Rivastigmine is a competitive, pseudoirreversible inhibitor with a half-life of 2 h, although it acts for approximately 10 h. The Adas-Cog scale and the CIBIC-Plus improve in patients treated with a daily dosis of 6 or 12 mg taken in two doses. Administration should be initiated at low doses (3 mg/day) which are progressively increased. Metriphonate is a prodrug of short life which inhibits acetylcholinesterase through a metabolite (DDVP) with a half-life in the circulation of 2 h. Improvement is observed in the ADAS-Cog scale and the CIBIC Plus and in behavior disorders at doses of 0.3 and 0.65 mg/kg/day. Doses between 30 and 60 mg/day are effective. CONCLUSIONS: Different studies carried out with the acetylcholinesterase inhibitors donepezil, rivastigmine and metriphonate have been effective in the control of the cognitive symptoms of Alzheimer disease in the initial or moderate phases of the disease. PMID- 10363494 TI - [Primary orthostatic tremor]. PMID- 10363495 TI - [Symptomatic paroxysmal dystonia (non-kinesigenic forms): two new cases]. AB - We report two new cases of symptomatic paroxysmal non-kinesigenic dystonia. The first is a 68-year-old woman with paroxysmal spontaneous dystonic spasms in her right arm lasting 1 minute. They occurred 1-2/day, a few months after a cerebral infarction (left internal capsule and left lenticular nucleus) which occurred 6 years ago. The second is a 30-year-old woman with a 7-year-history of spontaneous dystonic postures (flexion spasms) in her left arm lasting 15 minutes and occurring monthly. In this case an Arnold-Chiari malformation with cervical syringomyelia was discovered. PMID- 10363496 TI - [Localized continuous muscular activity secondary to radioculopathy S1 with shin hypertrophy and focal myositis]. PMID- 10363497 TI - [Proteasone, ubiquitine and neurodegenerative diseases]. PMID- 10363498 TI - It's a matter of life or death. PMID- 10363500 TI - Necessity is... PMID- 10363499 TI - A legal balancing act. PMID- 10363501 TI - Reconstruction of the severely atrophic maxilla in a young adult with periodontosis. AB - Reconstruction of the atrophic maxilla in a young adult presents unique challenges. This article describes reconstruction in a 30-year-old patient using a two-stage procedure. Bilateral maxillary sinus augmentations with simultaneous corticocancellous grafting to the anterior maxilla and alveolar ridge were performed. Eight endosseous implant fixtures were subsequently placed into the maxilla after a five-month healing phase. Six months later the implants were uncovered, healing abutments placed and a maxillary vestibuloplasty performed. The rationale behind this treatment and a review of the literature are discussed. PMID- 10363502 TI - The experience of a senior dental student in diagnosing pathology. AB - Dental school affords the opportunity to use various dental sciences, that is, radiology, pathology, histology, oral and maxillofacial surgery, in determining normal and pathological conditions. The case presented here describes the diagnosis, treatment and management of a rare intraosseous lesion, calcifying epithelial odontogenic tumor (CEOT), from the perspective of a senior dental student. PMID- 10363503 TI - Bone marrow transplants. Current applications & implications for oral health. AB - The application of bone marrow transplant (BMT) therapy to address a variety of pathologies has increased dramatically in the last decade. The list of diseases treated by this complex technology is quite lengthy. Side effects of BMT include a variety of documented untoward effects on oral health, many of which are age dependent. With the increasing number of people, particularly children, receiving bone marrow transplants, it is entirely possible that these recipients may appear as "routine, healthy" patients in a general practice or other oral health care setting. They will present significant, unusual findings related to their history of BMT treatment. The purpose of this paper is to review this treatment modality, its current applications, and the short- and long-term effects that the oral health care practitioner must identify, understand and address. PMID- 10363504 TI - Current and future approaches for diagnosis of periodontal diseases. AB - Our understanding of the etiology and pathogenesis of periodontal diseases has grown in the recent past, and new findings have led to advances in patient management. This article summarizes important new knowledge and offers a description of traditional and novel diagnostic approaches. These include clinical and radiographic assessments of the periodontal tissues, evaluation of the microbial challenge and the host response, and certain elements of the host genotype that may confer susceptibility to destructive periodontal diseases. PMID- 10363505 TI - UB researchers identify oral bacteria likely to trigger heart attack. PMID- 10363506 TI - [The molecular epidemiology of adenovirus]. PMID- 10363507 TI - [Comparison of stromal remodeling and keratocyte response after corneal incision and photorefractive keratectomy]. AB - PURPOSE: We investigated the keratocyte response and stromal remodeling after corneal incision and photorefractive keratectomy, respectively to learn the difference between the two surgeries histophysiologically and immunohistochemically. METHODS: We performed corneal incision or photorefractive keratectomy in rabbits or rats, and then we chronologically observed the histological changes and the changes in localization of extracellular matrix proteins. RESULTS: In both types of surgery, the keratocyte population in the damaged stroma became sparse, and the cells were undergoing apoptosis immediately after the procedures. After that, activated keratocytes migrated into the acellular zone, and the cells formed multiple layers at the resurfaced subepithelial regions. Deposition of amorphous substances was observed between migrated keratocytes, and stromal remodeling began. Three months after the surgery, corneal structure had recovered to near normal condition in the corneal incisions. In photorefractive keratectomy, however, strong immunoreactivity of extracellular matrix proteins was observed in the subepithelial regions. CONCLUSIONS: These results suggested that stromal wound healing processes were similar in both corneal incision and photorefractive keratectomy. Corneal incision may induce transient keratocyte response during the stromal remodeling, but photorefractive keratectomy may induce sustained keratocyte response. PMID- 10363508 TI - [Morphological changes in rabbit corneal endothelium after surgical injuries]. AB - PURPOSE: To understand the responses of the corneal endothelium to different types of surgery, we chronologically investigated the morphologic changes in the endothelial cells. METHODS: We did a mechanical incision, epithelial ablation, and excimer laser radiation on rabbit corneas and observed the morphologic changes in the endothelial cells for up to 2 weeks after surgery under a light microscope and an electron microscope. RESULTS: Although we observed enlarged intercellular spaces between neighboring endothelial cells, intercellular adhesion kept the cells tightly joined near their apexes immediately after each procedure. No signs of endothelial cell degeneration were observed after the procedures, but we did observe many Golgi apparati, rough-surfaced endoplasmic reticula, and secreted granules, indicating that the cells had been activated. After each procedure, the intercellular junctions and spaces required different amounts of time to return to normal. CONCLUSIONS: These results suggest that the different kinds of surgical injuries affected the corneal endothelium but that the changes were reversible. PMID- 10363510 TI - [Effects of local administration of interferon-beta on proliferation of the retinal pigment epithelium]. AB - PURPOSE: We demonstrated effects of local administration of human interferon (IFN)-beta on the repair process of the rabbit retinal pigment epithelium (RPE). MATERIAL AND METHODS: We used adult pigmented rabbits in this experiment. We measured IFN-beta levels in the ocular tissues after sub-Tenon administration of human IFN-beta by means of enzyme-linked immunosorbent assay (ELISA) methods. Laser photocoagulation in moderate intensity was applied after IFN administration. The repair process of the RPE in laser lesion sites was examined histopathologically. RESULTS: Locally administrated IFN spread by diffusion into the intraocular tissues. The highest IFN level was detected in the choroid. In eyes treated with IFN, RPE cells proliferated vigorously to the center of the photocoagulated lesions on early stages after laser photocoagulation. Proliferation of RPE cells after laser photocoagulation was remarkable in eyes treated with large amounts of IFN. CONCLUSION: It was demonstrated histopathologically that sub-Tenon administration of IFN-beta promoted proliferation of RPE cells during the repair process after laser photocoagulation. PMID- 10363509 TI - [The effect of neurotransmitters on rabbit cornea]. AB - PURPOSE: To ascertain the effect of neurotransmitters added to the culture medium of rabbit corneal epithelium and stromal cells. METHOD: The corneal epithelium and stromal cells were cultured in RCGM medium. Three neurotransmitters were added to the medium : substance P, acetylcholine, and vasoactive-intestinal peptide (VIP). RESULTS: Proliferation of epithelial cells significantly increased after incubation for 24 hours with substance P (p < 0.05). There was no change in proliferation after addition of acetylcholine or VIP. The extension of epithelial cell layer increased after addition of substance P but not after addition of acetylcholine or VIP. No change was induced in proliferation of stromal cells or extension of the stromal cell layer after addition of any one of the three substances. CONCLUSION: Substance P stimulates the proliferation of corneal epithelial cells when added to the culture medium. PMID- 10363511 TI - [Vitreoretinal tomography and foveolar traction in macular hole development and macular pseudohole]. AB - OBJECT: To clarify the morphologic features and foveolar traction in macular hole development and macular pseudohole. METHODS: The vitreoretinal tomography of idiopathic macular holes and macular pseudoholes was observed with optical coherence tomography (OCT). RESULTS: In stage 1 and 2, foveolar intraretinal splitting was evident. The posterior hyaloid membrane was detected in 7 of 10 eyes with stage 1 and 2 holes. Dome-shaped vitreoretinal separation was seen in 6 of 7 eyes in which the posterior hyaloid membranes were detected. A complete posterior vitreous detachment was seen around a stage 3 hole. In the macular pseudohole with preretinal membrane, there was anterior and central displacement of the inner retina in the perifoveal region, resulting in a U-shaped deformation of the macular lesion. In contrast, the structure of the central fovea was virtually intact. CONCLUSIONS: On the basis of the OCT findings, the intraretinal splitting and the cyst formation are important features in the development of a macular hole. The dome-shaped vitreoretinal separation in the early stages of macular holes suggests that the posterior hyaloid membrane may not be taut but slack, and would not cause a tractional force continuously even in early-stage macular holes. PMID- 10363512 TI - [Effectiveness of eyeglasses for protection against ultraviolet rays]. AB - PURPOSE: The relationship between eyeglass size and protection of the eye surface from the effects of solar ultraviolet (UV) rays was investigated. METHODS: Solar UV rays irradiating the eye surface were measured on a mannequin which modeled the standard facial bone structure of a Japanese female. UV sensor chips (photo sensitivity: 260-400 nm) were attached to the ocular surface of the lid fissure. UV measurement was done from 12:00 to 15:00 on a sunny day in March. UV intensity was measured under the following conditions: 1) with or without eyeglasses, 2) wearing sunglasses with side protectors, and 3) wearing a cap with a 7 cm brim. Eyeglasses of four frame sizes (width: 48-57 mm) were put on the mannequin. All lenses were made of plastic and coated so as to be impervious to rays shorter than 400 nm. The refractive power was 0 diopters. At the same time, UV irradiation intensity from all directions (excluding from the earth direction) was measured using a polyhedron type UV sensor with 25 sensor chips. RESULTS: Except for eyeglasses with the smallest frame size, eyeglasses effectively reduced UV exposure to sunlight from the upper front direction. However, protection against rays from the upper temporal direction was extremely poor. Sunlight from the upper back was reflected by the posterior surface of the eyeglasses and reached the eye surface. CONCLUSION: The efficacy of eyeglasses against UV depends on their size. The shape of the eyeglasses and reflection from the posterior lens surface are also of great importance. Small eyeglasses do not offer ideal UV protection for the Japanese face shape. PMID- 10363514 TI - [Influence of myopic disc shape in a classification program of the Heidelberg retina tomograph]. AB - PURPOSE: We investigated the influence of myopic disc shape on the diagnostic capability of a glaucoma diagnostic software (classification program) of the Heidelberg retina tomograph (HRT). SUBJECTS: 66 eyes of 66 normal subjects and 78 eyes of 78 patients with open-angle glaucoma were studied. The criterion of glaucoma was a visual field defect appearing between Aulhorn classification stage II and stage V regardless of the maximum intraocular pressure value. METHODS: The subjects were divided into eyes with a myopic disc and those with a non-myopic disc on the basis of stereo disc fundus photographs without considering the refractive errors. Agreement between the classification program and the clinical diagnosis was evaluated by the calculation of sensitivity, specificity, and diagnostic precision, and the influence of the disc shape on HRT disc shape parameters was also evaluated. RESULTS: Sensitivity, specificity, and diagnostic precision of the classification program were 83%, 95%, and 89% in the eyes with a non-myopic disc, and 71%, 96%, and 83% in the eyes with a myopic disc, respectively. Rim volume, height variation contour, mean RNFL (retinal nerve fiber layer) thickness, and RNFL cross section area were significantly larger in the eyes with a myopic disc than in those with a non-myopic disc regardless of the clinical diagnosis. CONCLUSION: The classification program should be modified to adjust to a myopia-like disc shape in order to improve the capability of the glaucoma predictive diagnosis. PMID- 10363513 TI - [Genome analysis with restriction endonucleases recognizing 4- or 5-base pair sequences of adenovirus type 4]. AB - PURPOSE: To ascertain the chronological change of subgenome types of adenovirus type 4 (Ad 4) through DNA analysis. MATERIALS AND METHODS: We evaluated sixteen Ad 4 strains from patients with acute viral conjunctivitis at four eye clinics in Sapporo, northern Japan. Seven strains were obtained from December 1993 through March 1994 (first period). Nine strains were obtained from March through May in 1995 (second period). These strains were analysed using DNA restriction endonucleases recognizing 4- or 5-base-pair sequences, Taq I and Hin f I. RESULTS: The genome type of sixteen strains was Ad 4 a. Seven strains in the first period showed the same subgenome type. Nine strains in the second period showed two subgenome types: five strains showing the same subgenome type as in the first period, and four showing a different subgenome type. The two subgenome types were similar in DNA pattern, suggesting that both were derivatives of a common subgenome type. All the sixteen strains were a new subgenome type which was different from those previously isolated during 1985-1989. CONCLUSION: The subgenome types of Ad 4 in 1993-1995 have changed from those in 1985-1989. PMID- 10363515 TI - [Causes of visual field defects after vitrectomy]. AB - PURPOSE: An inferotemporal visual field defect sometimes occurs following vitreous surgery for idiopathic macular hole. There is a possibility that this visual field defect is due to damage to the superonasal retina by fluid or air irrigation through an inferonasal infusion port. We tested this hypothesis by placing the infusion port in the inferonasal sector during vitreous surgery. CASES AND METHOD: We performed vitreous surgery on 31 eyes with idiopathic macular hole. The infusion port was placed in the inferonasal sector. The vitreous cavity was replanced either by 20% SF6 or 12% C3F8. We did not abrade the retinal pigment epithelium within the hole. The visual field was assessed before and 1 month after surgery using a Goldmann perimeter. FINDINGS: Three eyes developed a wedge-shaped visual field defect in the inferonasal sector. No visual field defect developed in the other 28 eyes. CONCLUSION: The findings show that visual field defect following surgery for idiopathic macular hole is dependent upon the site of the infusion port. We presume that the visual field defect is consequent to retinal damage caused by the flow of air or fluid during surgery. PMID- 10363517 TI - Ultrasonographic evaluation of tumorous lesions in digital vessels. AB - Ultrasonography has recently been used for evaluation of various conditions in Orthopaedics. Ultrasonographic examination is a noninvasive screening test especially for soft tissue masses. Ultrasonography is also a useful and essential diagnostic tool in cardiovascular disorders because real-time images of heart and vessels can be obtained. However, there have been few reports which describe ultrasonographic evaluation of tumorous lesions in digital vessels. In this paper, such lesions in two cases were evaluated by ultrasonography. An aneurysm of the digital artery is one of the definite candidates for ultrasonographic evaluation. PMID- 10363516 TI - [Hemodynamics of prepapillary vascular loop in hemi-central retinal vein occlusion]. AB - BACKGROUND: It has been shown, by indocyanine green (ICG) videoangiography, that the prepapillary vascular loops in chronic central retinal vein occlusion (CRVO) serve as an anastomosis between the retina and the choroidal venous systems. Similar vascular loops may develop in hemi-central retinal vein occlusion (hemi CRVO). CASES AND METHODS: I performed indocyanine green and fluorescein angiography using a scanning laser ophthalmoscope in 8 eyes of 8 patients with hemi-CRVO to evaluate the angioarchitecture and hemodynamics of the retinochoroidal circulation. All eyes had a prepapillary vascular loop and were identified as ischemic. The patients' ages ranged from 28 to 77 years (mean, 57 years). The interval between onset of hemi-CRVO and angiography ranged from 2 to 42 months (mean, 13 months). Scattered laser photocoagulation had been applied to the affected area in all 8 eyes and vitreous surgery in one eye. RESULTS: Delayed dye flow in the affected retinal veins was present in all 8 eyes. The blood in the affected retinal veins drained through the prepapillary vascular loop into the branch or trunk of the intact central retinal vein. No anastomoses were seen between the vascular loop and the choroidal veins. The prepapillary vascular loop became more dilated during follow-up in one eye. CONCLUSIONS: The prepapillary vascular loop in hemi-CRVO served as a collateral from the affected into intact retinal veins and not into the choroidal venous system. PMID- 10363519 TI - [Tumors of the pancreas]. PMID- 10363518 TI - [A case report of perforated early gastric cancer]. AB - An 83-year-old woman was seen at the First Department of Medicine Fukui Medical School because of upper abdominal pain. A simple chest film taken in the upright position revealed free air under the diaphragm. She was referred to the department and underwent a laparotomy with a diagnosis of acute panperitonitis due to a perforated gastric ulcer. At laparotomy, there was a perforation measuring 5 x 5 mm at the anterior gastric body of the lesser curvature, covered with abdominal wall. Billroth I gastrectomy was carried out with no lymph node dissection. The histological examination of the surgical specimen showed early gastric cancer of type IIc invading the submucosal layer around the ulcer in the gastric wall, composed of well differentiated adenocarcinoma. The perforated early gastric cancers often present difficulty in diagnosis pre or intra operatively. So it is important to examine closely the resected specimen intra operatively and perform frozen section diagnosis whenever possible. Forty-six cases of perforated early gastric cancer collected from the Japanese literature are also discussed. PMID- 10363520 TI - Duodenal obstruction by gallstone: case report of Bouveret's syndrome. AB - Bouveret's syndrome involves gastric outlet obstruction by gallstone. Herein we describe an unusual case of duodenal bulb obstruction by gallstone. An 80-year old woman was hospitalized with a fifteen-day history of vomiting. Computed tomography (CT) showed pneumobilia and a round calcified mass in the second portion of the duodenum. Upper gastrointestinal tract series demonstrated the same sized oval radiolucency between the bulbus and the second portion of the duodenum. Endoscopic examination revealed a round black mass in the second portion of the duodenum, totally occupying the lumen. Endoscopic removal and destruction of the gallstone was attempted using a dye-laser, but the stone was too hard to crush. Eventually surgical enterolithotomy was successfully performed without cholecystectomy or closure of the fistula. Improved preoperative systemic management and prompt examination allowed earlier surgical intervention and reduced the morbidity. Surgical approach whether fistula closure should be performed remains controversial. PMID- 10363521 TI - A patient with a traumatic right diaphragmatic hernia occurring 4 years after sustaining injury--statistical observations of a delayed diaphragmatic hernia caused by uncomplicated injury in Japan. AB - We describe our experience with a patient in whom a traumatic right diaphragmatic hernia developed 4 years after sustaining injury and review cases of delayed diaphragmatic injury reported in Japan. The patient was a 28-year-old man who sustained a severe contusion of the right epigastric region and fractured a right rib in a traffic accident in September 1992. In August 1996, the patient presented with shortness of breath on effort or after meals. A chest roentgenogram revealed intestinal gas in the right side of the thoracic cavity. A right diaphragmatic hernia was diagnosed on the basis of a gastrointestinal series, and the patient was operated on. The hernial orifice extended anteriorly from the central tendon in an 11:00 direction and measured 11 x 6 cm. The small intestine, right side of the colon, and liver were herniated. A total of 297 cases of blunt traumatic diaphragmatic hernia were reported in Japan between 1981 and 1996, including 47 cases (left side, 32 cases; right side, 15 cases) of delayed diaphragmatic hernia, defined as occurring one month or more after injury. Diaphragmatic hernia should be considered as a possible diagnosis in patients with abnormal shadows in the thoracic region who have recently sustained injury or who have a past history of injury. PMID- 10363522 TI - [Pathophysiology and management of diabetes mellitus in the elderly]. PMID- 10363523 TI - [Chemokines and inflammation]. PMID- 10363524 TI - [Mechanism of aging: obesity, non-insulin dependent diabetes mellitus (NIDDM) and abnormal behavior, and CCK-A receptor gene abnormality]. PMID- 10363525 TI - [The molecular mechanisms of estrogen action against senile disease]. PMID- 10363526 TI - [cDNA cloning of ju-myo protein (JP) and elucidation of the molecular mechanisms by which it exerts neurotrophic effects on neurons]. AB - The adult life span in inbred strains of Drosophila melanogaster (Dm) has been found to be controlled by a few major genes (Hereditas 111:207, 1989; Hereditas 117: 251, 1992). A 77 kDa protein, which we named ju-myo (life-span) protein (JP) and is supposed to be the product of the gene on autosomal locus JmA on adult Dm was shown to have life-span prolonging effect when it was supplied in food. However, the knowledge of its structure and molecular mechanisms by which JP exerts its effects on cells is still unclear. Here we show that JP can exert neurotrophic activities on postmitotic fetal rat neurons isolated from cerebral cortical and dopaminergic neurons isolated from the midbrain: it enhanced survival of MAP2-positive cells and tyrosine hydroxylase immunoreactive neurons by approximately 2-fold over the control group. JP did not increase the density of astrocytes nor expression of glial fibrillary acidic protein (GFAP) in the mesencephalic neuron cultures. Amino acid analysis of JP protein showed that JP is identical to the larval serum protein 2, of which sequence and structure were determined in 1997. Our work provides basis for defining the physiological role of JP at the molecular level and for exploring its potential utility as an alternative approach to study mechanisms of aging. PMID- 10363527 TI - [Gene therapy to treat Parkinson's disease]. AB - One recent strategy of gene therapy is to have cells express the lacking substances. Decline in dopamine D2 receptors (D2R) is observed in late-stage Parkinson's disease. We have constructed a replication-deficient adenovirus vector to transfer rat D2R cDNA (AdCMV.DopD2R) to the brain as a possible therapeutic strategy and a replication-deficient adenovirus vector to express nothing (AdCMV.Null) as a control. Using tissue culture cells infected with this vector, we detected D2 R cDNA by Northern analysis and receptor protein in membrane preparations as specific binding of the D2R ligand, [3H] spiperone. In vivo demonstration involved autoradiographic analysis of [3H] spiperone binding in rat striatum, D2R expression was amplified above normal concentrations in the injection site. We investigated the expression and functionality of the adenoviral vector. Comparative analysis of the autoradiographic images from the striatum injected with AdCMV.DopD2R and the contralateral striatum injected with a control vector, AdCMV. Null, in male rats indicated that D2R binding was increased by 40-60% on days 3 and 5 after injection, but then declined to baseline levels by day 21. When injected with apomorphine on days 3 and 7 after vector injection, experimental groups that had received unilateral striatal injections of AdCMV. DopD2R exhibited a distinct and significant laterality in rotational behavior. These results provide the first demonstration of an adenovirally mediated, intracerebral delivery of a functional neurotransmitter receptor. PMID- 10363528 TI - [Apolipoprotein J and Alzheimer's amyloid beta solubility]. PMID- 10363529 TI - [Pathologic evaluation of the main cause of death in Japanese centenarians]. AB - A total of 42 Japanese centenarians (9 males & 33 females) autopsied in Tokyo Metropolitan Geriatric Hospital during 22 years (1975-1996) were clinico pathologically examined to determine details of the main cause of death. The main cause of death of the 42 cases were sepsis (16 cases), pneumonia (14 cases), suffocation (4 cases), heart failure (4 cases), cerebrovascular disorder (2 cases) and malnutrition (2 cases). Most pneumonias were caused aspiration of foreign bodies, and the origins of sepsis were pyelonephritis (7 cases), biliary tract infection (3 cases), necrotic lesions of the intestine due to ileus, ischemia and pseudomembranous colitis (3 cases) and indwelling vein catheter (3 cases). Malignant neoplasms were observed in 16 cases (38%), and 5 of them had 2 or 3 lesions. Thus, the total number of lesions of malignant neoplasms were 22, as follows; colonic cancer (36%), urinary bladder cancer (14%), lung adenocarcinoma (9%), gastric cancer (9%), malignant lymphoma (9%) and others. However, none of these malignant neoplasms were directly related with the cause of death. All 42 centenarians died not of simple "senile decay", but due to diseases. PMID- 10363530 TI - [Morning blood glucose determination in the monitoring of metabolic control in type 2 elderly diabetic cases treated by oral hypoglycemic agents]. AB - Daily blood glucose profiles were measured in 163 Type 2 elderly diabetic cases to evaluate whether a fasting (before breakfast) or a post-prandial (after breakfast) blood glucose concentration is able to predict blood glucose values throughout the day. In the diet-treated alone group (n = 61), the percentage of daily blood glucose profiles having plasma glucose values less than the 08:00 hours (before breakfast) value were as follows: 59.0%, 32.8%, 59.0%, and 55.7% at 18.00 (before supper), 24.00, 03.00, 06.00 hours, respectively. In group treated by oral hypoglycemic agents (OHA) (n = 102), these were as follows: 45.1%, 26.5%, 52.9%, and 67.6%, respectively. In the OHA group, the mean plasma glucose value at 08:00 hours was significantly higher in patients with the lowest plasma glucose levels between 60-79 mg/dl than in patients with these levels between 80 99 mg/dl (103.7 +/- 19.6 vs 118.7 +/- 16.9 mg/dl, p < 0.01), but that at 10:00 hours was similar in the two groups (218.8 +/- 43.9 vs 214.5 +/- 40.1 mg/dl). In patients with lowest plasma glucose levels of between 60-99 mg/dl, the 08:00 hours value correlated positively with that of 24:00 (r = 0.40), 03.00 (r = 0.53), and the 06.00 hours value (r = 0.69), but no correlation was observed with the 18.00 hours value. On the other hand, the 10:00 hours value was not associated with these time-points values. Our results reveal that before breakfast plasma glucose values are more predictive of low blood glucose values in the night during sleep than after-breakfast blood glucose values, but do not predict low blood glucose values before supper in patients on OHA. PMID- 10363531 TI - [Long-term prognosis of patients with initial cerebral thrombosis and the MRI findings]. AB - To clarify the relationship between long-term prognosis of patients with stroke and their MRI findings, 103 patients with initial cerebral thrombosis, who survived more than three months after the ictus, were studied for five years. The mean age of 98 patients (T group), who were followed up completely, was 73.1 years-old and 65 were men. The age-matched controls consisted of two groups: 65 subjects, who had hypertension and/or diabetes without a history of stroke (R group), and 85 subjects, who had any hypertension, diabetes and stroke (N group). MRI findings were divided into six categories: 1) types of causative lesion, 2) grades of periventricular hyperintensity (none, rims/caps, patchy, diffuse PVH), 3) number of spotty lesions, 4) presence of silent infarction. 5) ventricular dilatation, and 6) extents of brain atrophy. Types of causative lesion were subdivided into 3 subtypes; infarction of the perforating artery territory (P type), infarction of the cortical artery territory (C type), and brainstem infarction (B type). The presence of vascular risks and dementia, and the extent of activity of daily living (ADL) were assessed. The P, C, and B types were identified by MRI in 46, 36, and 16 of the T group, respectively. Motor impairment, dementia, and an ADL status of complete dependence at discharge were also seen in 84, 44, and 22, respectively. In the T group, 33 patients died during five years, which resulted in a cumulative mortality rate of 33.7% and an annual mortality rate of 8.2%. Based on log-rank analysis, the survival rate of the T group revealed was significantly lower than those of the R and N groups. The recurrent rate in the T group (annual stroke recurrence rate was 4.0%) was higher than in the R and N groups, but stroke recurrence was not the cause of death and two thirds of deaths were due to aspiration pneumonia and/or asphyxia. Cox hazard regression analysis for death due to respiratory diseases showed that the hazard ratios of infarction, patchy PVH, and more than 4 spotty lesions were 8.87 (p < .001), 0.31 (p = .058), and 0.44 (p = .098), respectively. Compared to the survival group, rates of complete dependence in ADL, dementia, and brain atrophy were significantly higher in the death group with low incidences of the P type and patchy PVH, which indicated small vessel disease. These findings suggested that in patients with cerebral thrombosis, even in the chronic phase, care should be taken to prevent pneumonia and/or asphyxia due to bulbar palsy. Furthermore, no MRI findings were distinct predictors of long-term prognosis, although infarction based on the small vessel disease had rather good outcome in terms of respiratory disease. PMID- 10363532 TI - [An 88-year-old woman with symptoms of intoxication due to a small dose of digoxin]. AB - An 88-year-old woman was admitted to our hospital because of palpitations, dyspnea, orthopnea and appetite loss. On admission, small crackles were heard on her lower back, and her liver was swollen. Chest rentogenogram showed cardiomegaly (cardio-thoracic ratio 65.5%) and bilateral pleural effusion. Electrocardiograms showed atrial fibrillation with an average heart rate of 95 per minute. Echocardiography revealed mitral stenosis. Because the patient was considered to be suffered from heart failure due to mitral stenosis with atrial fibrillation, furosemide (20 mg per day) and digoxin (0.25 mg per day) was started. After digoxin had been raised to a dose of 0.50 mg per day because of sustained rapid ventricular response on the fourth hospital day, she complained of nausea and vomiting. Serum digoxin concentration was 2.55 ng/ml on the next day, and 1.08 ng/ml 96 hours after discontinuing digoxin. There was no complaint after digoxin was restarted with a dose of 0.05 mg per day. She complained of nausea again on the third day when the digoxin was raised to a dose of 0.083 mg in a blinded study. This observation indicates that digoxin intoxication could occur even in the smaller dose of digoxin than usual in the elderly. PMID- 10363533 TI - Interns: do you help 'em or haze 'em? PMID- 10363535 TI - Teaching nursing students to critically evaluate electronic fetal monitor tracings. AB - Evaluation of electronic fetal monitor (EFM) tracings is challenging for nursing students and novice perinatal nurses. Use of a standardized nursing process tool facilitates recognition of electronic fetal monitoring patterns and application of appropriate nursing interventions. Feedback from expert nurses and nursing instructors validates students' critical thinking skills. PMID- 10363534 TI - Maternal phenylketonuria: a new cause for concern. AB - The start of newborn screening for phenylketonuria (PKU) during the early 1970s has given rise to an increasing number of women who have been identified and successfully treated for the disease in childhood and are now preparing to have children of their own. Early detection and initiation of nutritional therapy before conception is key to a successful pregnancy outcome. Nurses who understand the pathophysiology, dietary limitations, and management of PKU in pregnancy can provide the care necessary for optimal maternal and neonatal health. PMID- 10363536 TI - Neonatal skin care: the scientific basis for practice. AB - OBJECTIVE: To review the literature addressing the care of neonatal skin. DATA SOURCES: Computerized searches in MEDLINE and CINAHL, as well as references cited in articles reviewed. Key concepts in the searches included neonatal skin differences; neonatal skin and care practices for skin integrity; neonatal skin and toxicity; permeability; and contact irritant sensitization. STUDY SELECTION: Articles and comprehensive works relevant to key concepts and published after 1963, with an emphasis on new findings from 1993 to 1999. One hundred two citations were identified as useful to this review. DATA EXTRACTION: Data were extracted and organized under the following headings: anatomy and physiology of the skin; physiologic and anatomic differences in neonatal skin; nutritional deficiencies; skin care practices; and care of skin breakdown. DATA SYNTHESIS: Newborns' skin is at risk for disruption of normal barrier function because of trauma. In light of available evidence about differences in neonatal skin development, clinical practice guidelines are suggested for baths, lubrication, antimicrobial skin disinfection, and adhesive removal. In addition, basic care practices are suggested for maintaining skin integrity, reducing exposure to potentially toxic substances, and promoting skin health beyond the neonatal period. Preventive care recommendations are made for reducing trauma, protecting the skin's immature barrier function, and promoting skin integrity. CONCLUSIONS: This review generated evidence with which to create a new and comprehensive practice guideline for clinicians. Evaluation of the guideline is under way at 58 U.S. sites. PMID- 10363537 TI - The effect of relaxation therapy on preterm labor outcomes. AB - OBJECTIVE: To examine the effect of relaxation on preterm labor outcome. DESIGN: Quasi-experimental, with women who experienced preterm labor randomly assigned to a control or experimental group. The experimental group was to do a daily relaxation exercise. A third group was added to the study: women who were originally assigned to the relaxation group but were unable to adhere to the daily practice. Final data were analyzed for three groups: control (n = 40), experimental (n = 44), and nonadherent (n = 23) participants. SETTING: Women were referred to the study from physician offices and a hospital-based obstetric triage clinic in the Northwest. PARTICIPANTS: Total sample was comprised of 107 women with singleton gestations, documented contractions with cervical change, and intact membranes. INTERVENTIONS: The experimental group was instructed in a progressive relaxation exercise. The participants were given tapes of the exercise and instructed to do it daily. OUTCOME MEASURES: Study outcomes included gestational age at birth, rate of pregnancy prolongation, and birth weight. RESULTS: The outcome variables were analyzed using analysis of covariance, with the preterm labor risk score entered as a covariate to compensate statistically for group differences. A positive response to the relaxation intervention was found: The experimental group had significantly longer gestations and larger newborns when compared to the control and nonadherent groups. CONCLUSIONS: Relaxation therapy made a difference in preterm labor outcome. Women who practiced relaxation had larger newborns, longer gestations, and higher rates of pregnancy prolongation. Given the low cost of the intervention, it should be offered to all women at risk for preterm labor. PMID- 10363538 TI - Transitioning preterm infants with nasogastric tube supplementation: increased likelihood of breastfeeding. AB - OBJECTIVE: To compare nasogastric tube and bottle supplementation as two means of transitioning preterm infants to breastfeeding within an established breastfeeding support program. DESIGN: Prospective, randomized controlled trial; mothers and health care providers, who were not blinded. SETTING: Metropolitan private regional perinatal center; 40-bed intensive-care nursery. PARTICIPANTS: Eighty-four preterm breastfed infants whose birth weight was 1,000-2,500 g. MAIN OUTCOME MEASURES: Rates of exclusive and partial breastfeeding at discharge from the intensive-care nursery, and at 3 days, 3 months, and 6 months after discharge. RESULTS: Compared with infants receiving bottle supplements, infants receiving nasogastric tube supplements were more likely to be breastfeeding at discharge and at 3 days, 3 months and 6 months, after adjusting for confounding variables. Odds ratios (confidence intervals = 95%) showed that the group receiving nasogastric supplements was 4.5 times (1.4 to 15) more likely to be breastfed at discharge and 9.4 times more likely to be fully breastfed (3.1 to 28.4). There were significantly fewer apnea and bradycardia episodes in the group receiving nasogastric supplements, although they had more episodes that required stimulation for resolution. Groups were not different with respect to length of hospitalization and infant weight at discharge. CONCLUSIONS: Using nasogastric tube supplementation during transition to oral feedings increases the likelihood of breastfeeding at discharge, 3 days, 3 months, and 6 months. This intervention requires a program with skilled personnel and an environment that allows the mother and infant to be in close physical proximity. Further study should investigate differences in the effects on maternal confidence, imprinting, and suck mechanism when preterm infants are bottle fed and breastfed. PMID- 10363539 TI - Impact of perinatal loss on the subsequent pregnancy and self: women's experiences. AB - OBJECTIVE: To describe the experience of a pregnancy after perinatal loss. DESIGN: Descriptive, open-ended responses to a self-completed questionnaire. SETTING: Questionnaires were distributed at a prenatal visit and completed in the office or at home. PARTICIPANTS: Seventy-two women who were 17 to 28 weeks pregnant, with a history of one or two perinatal losses. MAIN OUTCOME MEASURES: Themes that emerged from the women's responses to the questions. RESULTS: Three main dimensions, Past Pregnancy, Current Pregnancy, and Self constituted the overall framework for the themes of pregnancy anxiety, significant points in time, ways of coping, safe passage, social acceptance, binding-in, and grief and loss. CONCLUSIONS: Pregnancy after perinatal loss is characterized by guarded emotions, anxiety about this pregnancy, marking off the progress of the pregnancy in terms of fetal development and safety, and individual ways of coping to meet the tasks of pregnancy by seeking out or avoiding various behaviors. Women who have experienced perinatal loss would benefit from interventions to help them through these anxiety-filled pregnancies. PMID- 10363540 TI - Women's experiences in coping with abnormal Papanicolaou results and follow-up colposcopy. AB - OBJECTIVES: To delineate the primary concerns women associate with abnormal Papanicolaou (Pap) results and colposcopy and to identify women's strategies for coping with these potential stressors. DESIGN: A longitudinal, descriptive study involving telephone interviews after participants received abnormal Pap results and mailed questionnaires before and after colposcopy. SETTING: Private and public women's health clinics in the midwestern United States. PARTICIPANTS: Seventy-five women who had abnormal Pap results and needed initial colposcopy completed telephone interviews; 40 completed a precolposcopy questionnaire and 35 completed a postcolposcopy questionnaire. MAIN OUTCOME MEASURES: Clients' concerns and coping strategies were assessed after they learned their Pap results were abnormal, the day before colposcopy, and the day after the procedure. RESULTS: Women's primary concerns involved not understanding the Pap results, cancer, or infertility. Coping strategies used most, and rated as helpful throughout the experience, were seeking social support and distraction. CONCLUSIONS: Nursing interventions can be designed to improve women's understanding of the meaning of abnormal Pap results, address concerns about abnormal Pap results, and encourage women to use social support and distraction while awaiting colposcopy. PMID- 10363541 TI - Strategies for reducing the risk of malpractice litigation in perinatal nursing. AB - Perinatal nurses are involved in malpractice litigation most often as employees of a hospital being sued. Contemporary case examples from malpractice claims provide the foundation for examining how perinatal nurses can become the focus of such litigation. Increasing demand for individual nurse accountability, cost containment strategies that require nurses to broaden their scope of practice and to supervise unlicensed assistive personnel, increasing use of medical technologies, and the reality of compromised newborns and unexplained outcomes place perinatal nurses at risk for continued malpractice vulnerability. Specific strategies for risk reduction can be used by the individual nurse and the institution in relation to hospital policies and procedures, application of the nursing process, documentation, birth videos, and delegation of tasks to unlicensed assistive personnel. PMID- 10363542 TI - Professional accountability and legal liability for the team leader and charge nurse. AB - The rapid evolution in health care systems has significantly altered the roles and responsibilities of team leaders and charge nurses. Charge nurses and team leaders are responsible for promoting safe and effective patient care and maintaining high clinical standards in all settings. The Nursing Practice Act, state board of nursing administrative rules and regulations, and resources from professional organizations guide team leaders and charge nurses in these functions. Case law in recent medical malpractice actions also has affirmed the central role that team leaders and charge nurses play in preventing negative patient outcomes. The professional accountability and legal liability of team leaders and charge nurses in perinatal settings are examined. Effective strategies for reducing legal risk also are presented. PMID- 10363543 TI - Fetal surveillance and monitoring legal issues revisited. AB - The issue of obstetric litigation and electronic fetal monitoring (EFM) is revisited. The controversy in the medical literature that suggests that we are in an era fraught with both medical and legal dilemmas in the use of EFM is explored. The role of the nurse expert, the standard of care, and case studies are presented that demonstrate the need for obstetric nurses to be competent in EFM, the physiology and pathophysiology of labor, and the standard of care to which they are held accountable. Suggestions are made for risk management in the intrapartum setting. PMID- 10363544 TI - Legal issues in neonatal nursing: considerations for staff nurses and advanced practice nurses. AB - A neonatal nurse is a professional with special training, skill, and knowledge in the care of newborns and their families. The neonatal nurse is accountable to the patient, profession, and employer. Failure of the neonatal nurse to meet these obligations can result in liability in the profession, liability in the employment, a civil suit, or a criminal conviction. Regardless of the health care setting, professional nurses, whether at the bedside or in advanced practice, are morally, ethically, and legally accountable for their nursing judgments and actions. Although most nurses assume they will never be named in a lawsuit, and it is true that few are, their professional actions can be the focus of a suit. An overview of the legal implications found within neonatal nursing practice is presented. Two recent legal cases are presented and discussed to illustrate neonatal nursing and advanced practice liability. PMID- 10363545 TI - Informed consent issues in assisted reproduction. AB - Given the enormous ethicolegal controversies surrounding the use of assisted reproductive technologies (ART) in the United States, the most important role for nurses may be helping couples and third party participants obtain fully informed consent. The high compensation fees for egg donors may place them at special risk of exploitation. New government and professional guidelines, broader representation on ethics committees, and expanded counseling about risks and benefits can help reduce the potential for litigation and enhance patient autonomy. PMID- 10363546 TI - Efficacy of microsurgical free-tissue transfer in chronic osteomyelitis of the leg and foot: review of 22 cases. AB - The efficacy of free composite tissue transfer for the treatment of chronic osteomyelitis of the leg and foot was evaluated in a retrospective study. Twenty two patients, operated on at the American University of Beirut between January, 1992 and December, 1996, were identified. Infection involved the heel (8), ankle (1), foot (5), and tibia (8). All patients had multiple debridement and prolonged antibiotic treatment prior to presentation. The mean duration of disease was 4.8 years (range: 1 to 25 years). There were five cases of infected tibial non-union and one case of an infected tibial bone defect measuring 15 cm. Following radical debridement, microvascular free-tissue transfer was immediately performed. One latissimus dorsi and 13 rectus abdominis muscle flaps, as well as eight radial forearm fasciocutaneous flaps were used. At a mean follow-up of 3.8 years, there was one rectus abdominis free-flap failure in a Gustilo IIIC tibial fracture, which necessitated secondary amputation; there was no evidence of recurrence of osteomyelitis in the remaining 21 patients during the study period. The patients with tibial nonunion and bone defect healed following resection and bone transport utilizing a callus distraction technique. The results show that free tissue transfer is a safe and viable treatment option in chronic osteomyelitis of the leg and foot. A brief discussion of the history of microvascular free-tissue transfers, as well as their value in modern reconstructive surgery, is also presented. PMID- 10363547 TI - Intravenous bolus infusion of heparin for circulatory insufficiency after finger replantation. AB - When heparin 2,500 to 5,000 U was administered by intravenous bolus infusion to 13 fingers that had developed arterial thrombosis after replantation, complete survival was achieved in seven of 13 fingers. If fingers demonstrating partial necrosis were included, survival was obtained in 11 of 13 fingers. In nine fingers in which survival was obtained and follow-up observation was possible, sensory recovery was favorable, and there was no limitation in the use of the affected fingers, although mild atrophy of the pulp and nail deformity were noted in fingers with partial necrosis. Using a model of arterial thrombosis created in the carotid arteries of Sprague-Dawley rats, the authors investigated how the intravenous bolus infusion of heparin affected thrombolysis. During the early phase of arterial thrombosis when the thrombus is fragile, the intravenous bolus of heparin affected the balance between coagulation and fibrinolysis, facilitating the thrombolytic process, and increasing the rate of recanalization of the occluded arteries. Since an intravenous bolus injection of heparin is an easy procedure, without the risk of any severe side effects, this method should be considered in such cases of arterial thrombosis, with attention paid to the general condition of the patient. PMID- 10363548 TI - Microsurgical reconstruction of extensive scalp defects. AB - Large, full-thickness scalp defects represent a reconstructive challenge that has benefitted greatly from the introduction of microsurgical techniques. The authors review their experience with 16 patients with acquired defects of the scalp for which local or regional reconstructive options were unavailable. The mean age at the time of operation was 44.8 years. Nine patients underwent resection of malignant scalp lesions, followed immediately by free-flap coverage. Six patients required revision procedures for unstable scar as a result of prior trauma (2), old scalp avulsions (2), and multiple intracranial procedures (2). The remaining patient underwent replantation of an acutely avulsed scalp. The free-flap donor sites utilized included latissimus (6), scapular (3), radial forearm (2), rectus abdomnis (2), and omentum (2). Vein grafts were required in four cases. All flaps survived, although one required anastomotic revision and skin grafting for superficial loss. Additional complications were limited to seromas at two latissimus donor sites. Tumor control rates were poor, with all malignancy associated defects having persistent disease or recurring soon after surgery. All patients eventually achieved full defect coverage. The authors conclude that microsurgical reconstruction is a reliable option for providing stable coverage of large, complex, scalp defects. PMID- 10363549 TI - Reconstruction of accessory nerve defects with vascularized long thoracic vs. non vascularized thoracodorsal nerve. AB - Modern techniques of lymph-node neck dissection aim at conserving the accessory nerve. However, its continuity cannot be retained in cases of tumor in its direct neighborhood. In these cases, the accessory nerve must be resected for oncologic reasons. This study reports on neuronal reconstruction with both a vascularized long thoracic nerve transfer and a free thoracodorsalis nerve transfer, and compares the two. Both nerve transfers were removed simultaneously with an osseo myocutaneous scapula-latissimus dorsi transfer. In both cases, morphologic reconstruction in the face and a neuro-functional reconstruction of the shoulder arm region is possible. The vascularized long thoracic nerve transfer was superior to the non-vascularized throacodorsalis transfer for patients who had undergone radiotherapy. It resulted in more rapid healing and an improved motor result in shoulder elevation and maximal arm abduction. The long thoracic nerve transfer should thus be favored in reconstruction of the accessory nerve following tumor resection. PMID- 10363550 TI - Denervation of Pacinian corpuscles: electron microscopic observations in the rat following nerve transection. AB - Nerve transection using rat sciatic nerves was employed to observe morphologic changes in the periodic denervation of Pacinian corpuscles. During periods of from 1 to 20 weeks after surgery, a total of 15 corpuscles were obtained under the operating microscope and processed for morphologic analysis using light and electron microscopes. Based on the morphologic findings, normal corpuscles were composed of an axon terminal and inner and outer core cells. Following nerve transection, the axon terminal immediately disappeared at 1 week, but the original inner and outer core cells were preserved, with macrophage migration occurring in the outer core. After 8 weeks, the circular regularity of the inner and outer core lamellae was occasionally broken. The lamellae had wavy courses and there were many empty spaces in the inner core. Even at 20 weeks after denervation, there was interruption in the continuity of the outer core cells; the membranous structure of the outer core and the lamellar structure of the inner core were well-preserved; the outer and inner core cells were still present. Simultaneously, a small number of collagen fibrils were observed between the inner and outer core cells from the earliest postoperative stage. These fibrils increased in number in later stages. The results suggest that, immediately after destruction of axon terminals, macrophages migrate into the interlamellar spaces where they engulf the debris of the degenerated axon terminals. The degeneration of the corpuscles seems to be similar to that of denervated nerve axons; therefore, the inner core cells, which are continuous with the Schwann cells, maintain their original condition for at least 20 weeks after corpuscle denervation. The outer core cells, which are continuous with the perineurial nerve cells, are assumed to be the main cells producing the collagen fibrils in the denervated Pacinian corpuscles. PMID- 10363551 TI - Schwann cells can induce collateral sprouting from intact axons: experimental study of end-to-side neurorrhaphy using a Y-chamber model. AB - Axonal regeneration across end-to-side neurorrhaphy has recently been reported; however, neither the mechanism by which collateral sprouting from intact axons is elicited, nor the origin of the regenerating axons are known. There has even been controversy over the presence of collateral axonal sprouting from intact axons altogether. This reported experimental study was designed to clarify these questions. A rat sciatic nerve model was used. To avoid any mechanical damage to the donor nerve during the procedure, a Y-shaped silicone chamber was employed instead of direct suture. Axonal regeneration from the intact tibial nerve across the gap into the peroneal nerve was assessed using a retrograde neurotracer and immunohistochemical staining. Axonal regeneration across the gap was observed in 66 percent of the animals. The neurotracer evaluation clearly showed that all regenerating axons were sensory axons from the dorsal root ganglia. The authors concluded that Schwann cells from the distal wallerian degeneration of nerve segments did elicit collateral axonal sprouting from intact sensory axons, but not from motor axons in end-to-side neurorrhaphy. Invasion of the Schwann cells into the epineurial layer was the crucial step for the initiation of collateral axonal sprouting from the intact axons. PMID- 10363552 TI - Strain differences in peripheral-nerve regeneration in rats. AB - Currently, several strains of rats are used for studies of peripheral-nerve injury and repair. The purpose of this study was to determine if significant differences in regeneration between strains exist that might influence comparison of results and interpretation of scientific conclusions. One outbred (Sprague Dawley) and four inbred stains (ACI, Wistar-Furth, Lewis, Brown-Norway) were studied. Animals were randomized to one of two experimental conditions, undergoing either posterior tibial nerve transection and repair, or Silastic conduit repair of the posterior tibial nerve (n=6/group). Endpoint evaluations at 6 and 13 weeks included histomorphometry and walking-track analysis. Evidence of excellent regeneration was noted in all rat strains undergoing primary repair. Generally, no statistically significant differences between strains were noted, regardless of endpoint evaluation used in the primary repair group. Nerve regeneration across the conduits was either poor or not present at 6 weeks, with no regeneration at all noted in any animals in the ACI and Brown-Norway groups, and regeneration in only one or two animals in the other strains. At 13 weeks, between three and five animals in each strain showed regeneration, but functional recovery was poor. Overall, few differences in peripheral-nerve recovery appear to exist between rat strains. It seems that uniform conclusions may be drawn regardless of strain used. PMID- 10363553 TI - Effect of a new immunosuppressant, FTY720, on joint allografts. AB - The immunosuppressive effects of a new agent, FTY720, on joint allografts were studied histologically in a rat model. Favorable results without side effects were obtained at a dose of 3.0 mg/kg/day. The authors believe that this agent is one of the most useful immunosuppressants in the reported experimental model. PMID- 10363554 TI - Gracilis myocutaneous free-flap model in the rabbit. AB - This study was carried out to describe the anatomy and microsurgical free transfer of the gracilis myocutaneous flap in a rabbit model, and to evaluate its usefulness in further reconstructive microsurgical investigations. PMID- 10363555 TI - Shortening of rat teeth prevents autocannibalization of surgical flaps. AB - Self-biting in laboratory rats is an irritating problem that can impede progress and affect the credibility of experimental interventions. Autocannibalization frequently complicates the epigastric skin flap model that is often used to evaluate flaps. In one of the authors' studies, the autocannibalization rate of 55.6 percent in nine rats with unprotected skin flaps of the entire abdominal wall necessitated the application of previously reported plastic collars and protective vests to hinder self-biting. Because the use of these devices resulted in distress symptoms in nine of 11 rats with collars and flap congestion in five of five rats with vests, rat incisor teeth shortening was introduced as a stress free method to prevent chewing. Careful teeth cutting obviated autocannibalization in 19 of 20 rats (p<0.01) during a 5-day follow-up. Rat incisor teeth shortening performed every 3 or 4 days is a simple, inexpensive, and reliable method to prevent autocannibalization in the rat, while maintaining the well-being of the animal. PMID- 10363557 TI - Place therapies for older adults: conceptual and interventive approaches. AB - The study of place transitions-moves between places as well as changes occurring in environments of elders who age in place-is a relatively new, diverse research area of high relevance for adult developmentalists and gerontologists. This article explores the usefulness of a tractable model of environmental stress and proposes three potential "place therapies" that may minimize the negative impacts of place transitions upon older adults. Specifically, a transactional model of environmental stress linked to behavior setting theory is proposed for understanding both positive and negative outcomes associated with different kinds of place transitions. Three distinct "place therapies" are considered as interventions that may hold promise for preventing, ameliorating, and enriching the diverse impacts of place transitions on older adults and their environmental milieu. PMID- 10363556 TI - Cold preservation of nerve grafts decreases expression of ICAM-1 and class II MHC antigens. AB - Cold preservation has previously been shown to decrease the antigenicity of nerve allografts, while Schwann cells remain viable. The expression of intercellular adhesion molecule-1 (ICAM-1) and class II MHC antigens, both of which have been shown to play a major role in initiating graft rejection, was studied in fresh rat nerve, and after 2 and 7 weeks of cold preservation. Ten sciatic nerves harvested from Lewis rats were cut into three segments. One segment was processed immediately, while the other ones were preserved at 5 degrees C for 2 and 7 weeks, respectively, before processing. Immunostains using specific monoclonal antibodies and alkaline phosphatase development were performed on each sample. The relative level of expression of these antigens was compared using computer assisted densitometry. Expression of ICAM-1 was significantly decreased at 7 weeks, as compared to fresh and 2-week groups, with no statistically significant difference between fresh and 2-week nerves. Expression of class II MHC was significantly decreased at 2 and 7 weeks, compared to fresh nerves, with no statistically significant difference between the preserved groups. The decrease in antigenicity of cold-preserved nerve allografts appears to be linked to a down regulation of ICAM-1 and MHC class II expression. PMID- 10363558 TI - Elderly community residents' evaluative criteria and preferences for formal and informal in-home services. AB - This article focuses on the evaluative criteria of elderly community residents regarding their preferences in cases of long-term care decision-making. An overall picture of the evaluative criteria which the elderly use to evaluate various alternatives for long-term care are assessed. Furthermore, we determined which of these evaluative criteria may be considered as the most important by the elderly. A good relationship with informal carers appears almost pre-conditional to a preference for informal support. The desire not to burden acquaintances, as well as a positive previous experience with this type of care, are the most important reasons stated for choosing formal or private services. Insights into criteria that are used to evaluate different care arrangements clarify and refine our perspective on future developments. PMID- 10363559 TI - Custodial grandparenting: stresses, coping skills, and relationships with grandchildren. AB - This cross-sectional study compared three groups of custodial grandparents, those raising problematic grandchildren, those raising "normal" grandchildren, and noncustodial grandparents to identify the unique challenges and expectations faced by custodial grandparents due to their nontraditional roles while attempting to disentangle grandparental role demands from child-specific problems as sources of distress. Those grandparents raising grandchildren demonstrating neurological, physical, emotional, or behavioral problems exhibited the most distress, the most disruption of roles, and the most deteriorated grandparent grandchild relationships. Although custodial grandparents raising apparently normal grandchildren demonstrated less distress, less disruption of roles, and less deterioration of the grandparent-grandchild relationship than those grandparents raising grandchildren displaying problems, in these respects, they still demonstrated higher such levels than did traditional grandparents. PMID- 10363560 TI - Does public and private religiosity have a moderating effect on depression? A bi racial study of elders in the American South. AB - Religious activities are shown to correlate with rates of psychological depression symptoms in a sample of 995 African American and white elderly residents of Nashville. The data, collected in face-to-face interviews, included indicators of both public and private religiosity. Levels of religiosity and perceived social support were higher among the African-American respondents than among others, and among female respondents. Separate regression analyses of the racial groupings, which appeared to have distinctive religious subcultures, generally show that perceptions of social support mediate the relationship between levels of religiosity and symptoms of depression. PMID- 10363561 TI - Individual differences in aging minorities. AB - To fully understand the differences present between various ethnic and racial groups, there must be an understanding of the heterogeneity that is represented within a given ethnic/racial group. The purpose of this article is to discuss the importance of an individual differences approach in studying the ethnic diversity of an aging population. Conceptual, methodological, and design issues are discussed with the goal of better understanding the developmental processes of aging minority elderly populations. PMID- 10363562 TI - Stability of older women's friendships: a commentary on Roberto. PMID- 10363563 TI - Different expression patterns of nitric oxide synthase isozymes in various gynecological cancers. AB - The expression of nitric oxide synthase (NOS) in human gynecological cancers, including ovarian cancers, uterocervical cancers, and endometrial cancers for example, was examined by the reverse transcriptase/polymerase chain reaction, coupled with Southern hybridization and by immunohistochemistry. Nitric oxide synthase II (NOS II), an inducible form, was expressed in more than 90% of the cancers. Nitric oxide synthase I (NOS I), a neuronal form, was expressed in 58% of all the ovarian cancers, in which the serous type is found more frequently (5 out of 7) than the mucinous type (2 out of 6), and in all clear-cell cancers. The frequency of NOS I expression in uterocervical cancers and endometrial cancers was relatively low. Nitric oxide synthase III (NOS III), an endothelial form, was detected in 25% of ovarian and 33% of endometrial cancers, while no expression was detected in uterocervical cancers. In terms of cancer types, all clear-cell adenocarcinomas and most of the serous-type adenocarcinomas expressed both NOS I and NOS II, while most uterine squamous carcinomas and endometrial adenocarcinomas expressed only NOS II. However, there was no correlation between the frequency of NOS expression and patients' age or the clinical stage of the disease. Since NO increases vascular permeability and blood flow, the high frequency of NOS expression in gynecological cancers may serve to stimulate and promote tumor growth. PMID- 10363564 TI - Superoxide dismutases in the human colorectal cancer sequence. AB - PURPOSE: The oxidant-antioxidant balance within tissues is thought to contribute to the development and progression of cancer. Previous investigations have indicated changes in this balance during the colorectal oncogenic process that merit further investigation. The aim of the present study was to evaluate whether the human colorectal cancer sequence is accompanied by changes in the protein and activity levels of the antioxidant enzymes manganese- and copper/zinc-superoxide dismutase (Mn-SOD and Cu/Zn-SOD). PATIENTS AND METHODS: SOD levels were assessed in colorectal adenomas, carcinomas, and liver metastases and were compared with those in the corresponding normal tissues (n = 35 in each group). Mn- and Cu/Zn SOD expression was first evaluated semiquantitatively by electrophoretic activity analysis, immunoblotting, and immunohistochemistry and was subsequently quantified by enzyme-linked immunosorbent assays (ELISAs) and spectrophotometric activity assays. RESULTS: The semiquantitative analyses showed enhanced Mn-SOD levels, primarily localized in (neoplastic) epithelial cells, in carcinomas, and in liver metastases as compared with adenomas and normal mucosa, whereas no consistent pattern was observed for Cu/Zn-SOD. Normal liver tissue expressed the highest levels of both SODs. The quantitative SOD analyses confirmed these observations and revealed that carcinomas and liver metastases expressed 2-4 times more Mn-SOD protein and enzymatic activity (0.0005 < P < 0.01) than did the normal mucosa. Adenomas expressed intermediate Mn-SOD levels, which increased significantly with the diameter and tended to increase with the grade of dysplasia and presence of a villous component. In contrast, adenomas, carcinomas, and the corresponding normal mucosa were found to have a similar Cu/Zn-SOD content, whereas liver metastases contained significantly (P < 0.02) more Cu/Zn SOD as compared with these tissues. In addition, the Cu/Zn-SOD content was not related to any histopathological characteristic of the carcinomas or adenomas. CONCLUSIONS: Our study indicates that the development of neoplasia in the human colorectum is accompanied by major changes in the level and activity of Mn-SOD. This observation illustrates that Mn-SOD might have a functional role in human colorectal carcinogenesis. PMID- 10363565 TI - Characterization of a new potent, in vivo neutralizing monoclonal antibody to human vascular endothelial growth factor. AB - Vascular endothelial growth factor (VEGF) is an important mediator of tumor induced angiogenesis and represents a potential target for anticancer therapy. Therefore, we prepared a panel of monoclonal antibodies (mAb) against both the VEGF121 and VEGF165 isoforms. Three of them completely neutralized the mitogenic stimulation by VEGF of human umbilical vein endothelial cells at mAb concentrations below 0.1 microg/ml. The most potent one, with a dissociation constant (Kd) of 8 pM, inhibited, in a dose-dependent manner, VEGF-induced angiogenesis in a growth factor implant model in mice. A complete inhibition of the angiogenic response was obtained by daily intraperitoneal injections of 10 microg mAb/mouse. Angiogenesis induced by basic fibroblast growth factor was not inhibited by the mAb. Epitope mapping of the mAb, performed by competitive enzyme linked immunosorbent assay and Western blot analysis, showed that it did not bind to the reduced and denatured monomer of VEGF. Substitutions of three residues (Q87R, G88K, Q89K), located on the major surface loop beta5 to beta6 of VEGF, resulted in the complete loss of binding (more than 400-fold reduction). The results suggest that the mAb binds primarily to a conformation-dependent epitope on the VEGF dimeric form, encompassing one of the loop regions involved in KDR receptor binding. The mAb with its strong neutralizing properties represents a useful agent for effective blocking of VEGF-mediated tumor neovascularization. PMID- 10363566 TI - Diagnostic assessment of brain tumours and non-neoplastic brain disorders in vivo using proton nuclear magnetic resonance spectroscopy and artificial neural networks. AB - PURPOSE: Experiments were carried out to assess the potential of artificial neural network (ANN) analysis in the differential diagnosis of brain tumours (low and high-grade gliomas) from non-neoplastic focal brain lesions (tuberculomas and abscesses), using proton magnetic resonance spectroscopy (1H MRS) as input data. METHODS: Single-voxel stimulated echo acquisition mode (STEAM) (echo time of 20 ms) spectra were acquired from 138 subjects including 15 with low-grade gliomas, 47 with high-grade gliomas, 18 with tuberculomas, 18 with abscesses and 40 healthy controls. Two neural networks were constructed using the spectral points from 0.6 to 3.4 parts per million. In the first network construction, the ANN had to differentiate between tumours from infections, while the second network had to differentiate between all five histological classes. RESULTS: ANN analysis gave a histologically correct diagnosis for low- and high-grade gliomas with an accuracy of 73% and 98% respectively. None of the 62 tumours was diagnosed as an infectious lesion. Among the non-neoplastic lesions, ANN classification was correct in 89% of tuberculomas and in 83% of brain abscesses. The specificity of ANN diagnosis was 98%, 92%, 99%, and 100% for low-grade gliomas, high-grade gliomas, tuberculomas and abscesses respectively. CONCLUSION: The present data show the clinical utility of non-invasive 1H MRS by automated ANN analysis in the diagnosis of tumour and non-tumour cerebral disorders. PMID- 10363567 TI - Radiosensitizing effect of natural and recombinant beta-interferons in a human lung carcinoma in vitro. AB - PURPOSE: The enhancing effect of natural interferon-beta (n-IFN-beta) or recombinant interferon-beta (r-IFN-beta) on radiation damage in tumor cells has been evaluated in several sudies. It is not clear whether the different forms of IFN-beta available today are equally efficient in modulating the intrinsic radiosensitivity of tumor cells. The purpose of this study was to compare the radiosensitizing effect of n-IFN-beta, r-IFN-beta1a, and r-IFN-beta1b in one cancer cell line. METHODS: The A549 lung-cancer cell line was grown as a monolayer culture and incubated for 24 h with n-IFN-beta (Fiblaferon), r-IFN beta1a (Betaserin), or r-IFN-beta1b (Betaferon). Thereafter, the cultures were irradiated with single graded doses of 0, 1, 2, 4, and 6 Gy. Cellular survival was counted in a colony-forming assay at 10 days after treatment. Survival curves were established using the linear quadratic model. Statistical comparison of the survival data was performed using Student's t-test for each dose point. Interactions of IFN-beta and radiation were evaluated using isobologram analysis. RESULTS: All three types of IFN-beta enhanced the radiation sensitivity of A549 cells in a similar way as shown in the alteration of the survival curves and the isobologram analysis. The isoeffective concentration of r-IFN-beta1b was 2.7-fold higher than that of r-IFN-beta1a or n-IFN-beta. All three interferons increased the alpha-component of the survival curves in a concentration-dependent way, suggesting an influence on repair of radiation damage. The maximal sensitizing enhancement ratio (SER) obtained with n-IFN-beta or r-IFN-beta1a at 3000 IU/ml was 1.66 and 1.51, respectively. The highest SER, obtained with r-IFN-beta1b, at 8000 IU/ml was 1.93. CONCLUSIONS: All three interferons tested can equally modify the intrinsic radiosensitivity of A549 cells. The isoeffective concentration of r IFN-beta1b is 2.7-fold that of n-IFN-beta or r-IFN-beta1a. PMID- 10363568 TI - The influence of the p53 gene on the in vitro chemosensitivity of colorectal cancer cells. AB - PURPOSE: The p53 gene is considered one of the most important in the control of apoptosis, and its mutations have a close relationship with chemosensitivity. The aim of this work was to investigate the role of p53 in the apoptosis of colorectal cancer cells in vitro, induced by 5-fluorouracil (5-FU) and hydroxy camptothecin (HCPT). METHODS: A total of 39 colorectal cancer samples from patients were treated in vitro with 5-FU (10 microg/ml), 5-FU (10 microg/ml) + leucovorin (5 microg/ml), HCPT (0.1 microg/ml) and HCPT (0.1 microg/ml) + Salvia mitorrhiza (6 microl), using an in situ terminal deoxynucleotidyltransferase assay to detect chemosensitivity. p53 gene mutations from tumor DNA were detected, after amplification by the polymerase chain reaction of exons 5-8, by non-radioactive single-strand conformation polymorphism. RESULTS: p53 gene mutations were observed in 43.6% (17/39) of colorectal carcinomas, when the terminal deoxynucleotidyltransferase assay was used to detect the tumor apoptotic rate. Cells with mutated p53 had lower chemosensitivity than those without (p < 0.01). CONCLUSION: Routine assessment of p53 status may be helpful in selecting patients with the wildtype p53 gene, who have a predictably better response to chemotherapy. PMID- 10363569 TI - The influence of L-triiodothyronine, L-thyroxine, estradiol-17beta, the luteinizing-hormone-releasing hormone, the epidermal growth factor and gastrin on cell proliferation in organ cultures of 35 benign and 13 malignant human thyroid tumors. AB - PURPOSE: To characterize the influence of six factors on human thyroid tissues at the cell-proliferation level. These six factors were the epidermal growth factor (EGF), the luteinizing-hormone-releasing hormone (LHRH), triiodothyronine, thyroxine, estradiol and gastrin. METHODS: Forty-eight human thyroid specimens were obtained from surgical resection and maintained alive for 48 h ex vivo (in vitro) under organotypic culture conditions. These specimens comprised 35 benign cases (17 multinodular goiters and 18 adenomas) and 13 cancers. Cell proliferation in the control and treated conditions (at a 5 nM dose) was assessed by means of the thymidine labeling index, which enables the percentage of cells in the S phase of the cell cycle to be determined in accordance with autoradiographic procedures. RESULTS: The results show that, of the six factors tested here, EGF significantly (P < 0.05 to P < 0.001) increased cell proliferation in the greatest number of cancers as compared to what happened with the remaining five. Each of these six factors significantly increased or decreased proliferative cell activity in some 10%-30% of the cases under study. CONCLUSIONS: Triiodothyronine, thyroxine, LHRH and gastrin may increase or decrease cell proliferation in human thyroid tissues, whether benign or malignant, to the same extent as other hormones and/or growth factors such as thyrotropin, EGF, insulin-like growth factor 1, transforming growth factor beta1 and estradiol the effects of which on thyroid cell proliferation are already well documented in the literature. PMID- 10363570 TI - Results of a randomized double-blind placebo-controlled trial evaluating sequential high-dose cytosine arabinoside/mitoxantrone chemotherapy with or without granulocyte/macrophage-colony-stimulating factor in high-risk myelodysplastic syndromes. AB - A prospective, randomized, double-blind placebo-controlled trial was designed to evaluate the impact of granulocyte/macrophage-colony-stimulating factor (GM-CSF) on the efficacy of sequential high-dose cytosine arabinoside/mitoxantrone chemotherapy (S-HAM) in adult patients with high-risk myelodysplastic syndromes (MDS). GM-CSF or placebo was given subcutaneously once daily at a dose of 250 microg/m2, starting 48 h prior to chemotherapy, and continued until neutrophil recovery. Owing to high toxicity and slow patient recruitement the study was closed and unblinded after 31 patients had been enrolled; 15 were randomized to receive placebo and 16 to receive GM-CSF. A total of 29 patients were evaluable for response; their median age was 57 years. Ten patients achieved a complete remission (34.5%), 9 patients had persistent MDS (31%), 10 patients died within 6 weeks after the onset of treatment (early death) (34.5%). The median remission duration was 190 days (range: 2.5-45 months). Among the 29 evaluable patients no significant differences could be found between the two study arms regarding complete remission rate [GM-CSF: 31% (5/16) versus placebo: 38% (5/13) P = 0.45], rate of persistent MDS [GM-CSF: 25% (4/16) versus 38% (5/13) P = 0.35), early death rate [44% (7/16) versus 23% (3/13) P = 0.22] and remission duration (GM CSF: 87 days versus placebo 221 days). Duration of granulocytopenia (median: 33 days with GM-CSF) versus 35 days with placebo) and frequency of infectious episodes were not significantly influenced by GM-CSF. The small number of patients finally analyzed means that no definite conclusions about the effect of GM-CSF can be reached. PMID- 10363571 TI - Aging and cancer. The Eighth International Symposium of the Hiroshima Cancer Seminar, November 1998, Hiroshima, Japan. PMID- 10363572 TI - Lactoferrin inhibits hepatitis C virus viremia in patients with chronic hepatitis C: a pilot study. AB - Hepatitis C virus (HCV) is associated with the development of cirrhosis and hepatocellular carcinoma. We recently found that bovine lactoferrin, a milk protein belonging to the iron transporter family, effectively prevented HCV infection in cultured human hepatocytes (PH5CH8). We tested the hypothesis that lactoferrin inhibits HCV viremia in patients with chronic hepatitis C. Eleven patients with chronic hepatitis C received an 8-week course of bovine lactoferrin (1.8 or 3.6 g/day). At the end of lactoferrin treatment, a decrease in serum alanine transaminase and HCV RNA concentrations was apparent in 3 (75%) of 4 patients with low pretreatment serum concentrations of HCV RNA. However, 7 patients with high pretreatment concentrations showed no significant changes in these indices. This pilot study suggests that lactoferrin is one potential candidate as an anti-HCV reagent that may be effective for the treatment of patients with chronic hepatitis. PMID- 10363573 TI - Somatic mutation of the APC gene in thyroid carcinoma associated with familial adenomatous polyposis. AB - We report the existence of both germline and somatic mutations of the APC gene in thyroid carcinomas from familial adenomatous polyposis (FAP) patients. One papillary thyroid carcinoma from a 210-year-old woman, with germline mutation of the APC gene (TCA to TGA at codon 1110), showed a somatic mutation of AAAAC deletion between codons 1060 and 1063. Another somatic mutation of CAG to TAG at codon 886 was also found in one of multiple thyroid carcinomas from a 26-year-old woman with attenuated FAP and germline mutation at codon 175 (C deletion). This is the first evidence that total absence of the normal function of the APC gene is involved in development of thyroid carcinomas in FAP. PMID- 10363574 TI - Phytol is a novel tumor promoter on ICR mouse skin. AB - Phytol is a branched, long-chain aliphatic alcohol which has various biological effects. In this study, we examined phytol as a tumor promoter in a mouse skin initiation-promotion model, and compared its promotion activity with that of 12-O tetradecanoyl phorbol-13-acetate (TPA). Female ICR mice, 7 weeks of age, were initiated with 100 microg of 7,12-dimethylbenz(a)anthracene, and were then topically promoted twice a week for 16 weeks with 100 mg of phytol or with 2.5 microg of TPA. In this model 95% of animals treated with phytol developed skin tumors within 16 weeks. The average number of lesions per mouse treated with phytol was significantly lower than that in mice treated with TPA, and this significant difference continued up to 16 weeks after the end of promotion treatment. Characterization of hyperplasia 48 h after topical application of agents showed that epidermal thickness and vertical thickness following topical application of phytol were significantly increased compared with vehicle controls, but were significantly smaller than in animals treated with TPA. Ornithine decarboxylase (ODC) activity following topical application of phytol was increased in a dose-dependent manner and showed a weak, delayed induction (which was maximal 11-12 h after treatment) as compared with the case of TPA. The specific binding of [3H]phorbol-12,13-dibutyrate (PDBU) by JB6 cells was not inhibited by phytol at concentrations up to 1 mM. These results indicate that phytol has a weak tumor promoter activity compared to TPA and is a non-TPA-type tumor promoter in this model of mouse skin carcinogenesis. PMID- 10363575 TI - Role of copper accumulation in spontaneous renal carcinogenesis in Long-Evans Cinnamon rats. AB - Spontaneous renal cell tumors in totals of 223 male and female Long-Evans Cinnamon (LEC) rats of 51-120 weeks old, 157 male F344 rats of 51-120 weeks old, and 14 male Long-Evans Agouti (LEA) rats of 51-70 weeks old were examined histologically. The incidences of renal cell tumors increased with age in male and female LEC rats, but no tumors developed in F344 or LEA rats. Dilated atypical tubules of the kidneys were observed at high incidence in aged LEC rats. Copper staining of LEC rat kidneys showed a positive reaction in proximal tubules of the cortex and the outer stripe of the medulla. The renal copper concentration of LEC rats reached a peak in the period of necrotizing hepatitis with renal tubular necrosis, and was higher than that in F344 rats for up to 106 weeks. In contrast, the renal iron concentration of LEC rats was lower than that in F344 rats except in the period of necrotizing hepatitis. Long-term treatment of LEC rats with D-penicillamine, a copper-chelating agent, inhibited accumulation of copper, but not iron, in the kidneys, and inhibited the development of karyomegaly of proximal tubules and dilated atypical tubules. These results suggest that persistent copper accumulation after toxic necrosis of tubules is the major cause of spontaneous renal carcinogenesis in LEC rats. PMID- 10363576 TI - Effects of a soybean isoflavone mixture on carcinogenesis in prostate and seminal vesicles of F344 rats. AB - Several epidemiological studies have suggested an inverse association between the risk of prostate cancer and intake of soybeans and their products. In vitro data pointing to possible anti-carcinogenic properties of the soybean isoflavone, genistein, led us to investigate the chemopreventive potential of soybean isoflavones in a rat carcinogenesis model induced by 3,2'-dimethyl-4 aminobiphenyl (DMAB) and testosterone propionate (TP). Animals received DMAB s.c. injections at 2-week intervals for the first 20 weeks and implanted silicon tubes containing 40 mg of TP, replaced at 6-week intervals throughout the experiment. The soybean isoflavone mixture consisting of 74% genistein and 21% daidzein was mixed in basal diet (AIN-76A) at concentrations of 100 and 400 ppm and fed to F344 male rats throughout the experiment. Rats treated with carcinogens and administered isoflavone mixture at 100 and 400 ppm developed adenocarcinomas at incidences of 35% and 29%, respectively, in the prostate and seminal vesicles, whereas the figure was 60% for those maintained on control diet. Feeding of the isoflavone mixture at 100 and 400 ppm significantly inhibited the number of argyrophilic nucleolar organizer regions (AgNORs) in adenocarcinomas of the accessory sex glands as compared to those of rats fed control diet. No influence on the development of neoplastic lesions originating in other organs was noted. The results of this study provide evidence that soybean isoflavones may have potential as chemopreventive agents against carcinogenesis in the prostate. PMID- 10363577 TI - Organ-dependent modifying effects of caffeine, and two naturally occurring antioxidants alpha-tocopherol and n-tritriacontane-16,18-dione, on 2-amino-1 methyl-6-phenylimidazo[4,5-b]pyridine (PhIP)-induced mammary and colonic carcinogenesis in female F344 rats. AB - Modifying effects of caffeine, alpha-tocopherol, and n-tritriacontane-16,18-dione (TTAD) on 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP)-induced mammary and colonic carcinogenesis were investigated in female F344 rats. Groups of 20 rats, 6 weeks old, were given 0.02% PhIP (in diet) alone, or together with 0.1% caffeine (in drinking water), 0.5% alpha-tocopherol (in diet) or 0.1% TTAD (in diet) for up to 54 weeks. Groups of 10 females receiving basal diet or one of the test chemicals without PhIP supplementation were also maintained. The final combined incidences (adenomas plus adenocarcinomas) and multiplicity (No./rat) of mammary adenomas and adenocarcinomas were significantly lowered in the PhIP plus caffeine group (10%, 0.10) as compared to the PhIP alone value (40%, (1.50). Incidences of mammary tumors in the PhIP plus alpha-tocopherol or TTAD groups tended to be decreased while their multiplicities were significantly lowered. With regard to colon tumor development, on the other hand, rats given PhIP plus caffeine exhibited an elevated incidence (75% versus 15% in the control), whereas alpha-tocopherol and TTAD had no effect. Surprisingly, metabolic activation of PhIP was inhibited by addition of caffeine in an in vitro assay. The results indicate that caffeine exerts a potent chemopreventive action against PhIP induced mammary carcinogenesis, but acts as a co-carcinogen for PhIP-induced colonic carcinogenesis. PMID- 10363578 TI - Chemopreventive effect of N-(2-cyclohexyloxy-4-nitrophenyl)methane sulfonamide (NS-398), a selective cyclooxygenase-2 inhibitor, in rat colon carcinogenesis induced by azoxymethane. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) such as sulindac and indomethacin inhibit colon carcinogenesis, and selective cyclooxygenase (COX)-2 inhibitors are considered to be potential chemopreventive agents without the side effects of usual NSAIDs. We reported that NS-398, N-(2-cyclohexyloxy-4-nitrophenyl)methane sulfonamide, suppressed the formation of preneoplastic lesions, aberrant crypt foci (ACF), induced by azoxymethane (AOM) in a short-term assay of rat colon carcinogenesis. In this study, we examined the effects of long-term NS-398 administration on rat colon carcinogenesis. After three AOM treatments at weekly intervals, a dose of 10 mg/kg of NS-398 in 5% Arabic gum solution was administered by gavage three times per week in group 2 until the termination of the experiment. Rats in group 1 were fed in a basal diet and given 5% Arabic gum solution alone after AOM treatment. At 40 weeks after the first AOM treatment, all rats were killed and the whole intestines including colon were examined. While the incidences of whole intestinal and colon neoplasms in group 1 were 84.6% and 80.8%, respectively, those in group 2 (given NS-398) were 51.9% and 44.4% respectively (P=0.0177 and P=0.0103 by Fisher's exact test, respectively). The multiplicities in group 2 (0.67+/-0.78 and 0.48+/-0.58) were also decreased significantly compared with those (1.39+/-1.10 and 1.08+/-0.74) in group 1 (P<0.01 by Welch's method and P<0.002 by Student's t test, respectively). In immunohistochemistry for proliferative cell nuclear antigen (PCNA), the PCNA stained cell index (7.40+/-0.5) in group 2 was significantly decreased from that in group 1 (14.03+/-0.82) (P<0.001 by Welch's method). The results suggest that NS-398, a selective COX inhibitor, has a chemopreventive activity against colon carcinogenesis without side-effects such as gastric ulceration. PMID- 10363579 TI - PTEN/MMAC1 mutations in hepatocellular carcinomas: somatic inactivation of both alleles in tumors. AB - Allelic loss of loci on chromosome 10q occurs frequently in hepatocellular carcinomas. Somatic mutations of the PTEN/MMAC1 gene on this chromosome at 10q23 were recently identified in sporadic cancers of the uterus, brain, prostate and breast. To investigate the potential role of PTEN/MMAC1 gene in the genesis of hepatocellular carcinomas, we examined 96 tumors for allelic loss on 10q and also for subtle mutations anywhere within the coding region of PTEN/MMAC1 gene. Allelic loss was identified in 25 of the 89 (27%) tumors that were informative for polymorphic markers in the region. Somatic mutations were identified in five of those tumors: three frameshift mutations, a 1-bp insertion at codon 83-84 in exon 4 and two 4-bp deletions, both at codon 318-319 in exon 8; two C-to-G transversion mutation, both at -9 bp from the initiation codon in the 5' non coding region of exon 1. No missense mutation was observed in this panel of tumors. In most of the informative tumors carrying intragenic mutations of one allele, we were able to detect loss of heterozygosity as well. These findings suggest that two alleles of the PTEN/MMAC1 gene may be inactivated by a combination of intragenic point mutation on one allele and loss of chromosomal material on the other allele in some of these tumors. PMID- 10363580 TI - Increased choline kinase activity and elevated phosphocholine levels in human colon cancer. AB - Nuclear magnetic resonance spectroscopy has detected elevated phosphocholine levels in human tumor tissues and cells, and in cells that were transformed with the activated Ha-ras gene and stimulated in vitro with growth-promoting factors such as platelet-derived growth factor, epidermal growth factor, and phorbol ester. However, the mechanism of the elevation and the function of the increased phosphocholine levels have not been clearly demonstrated. We studied phosphocholine levels enzymatically and analyzed the activity of choline kinase, which catalyzes the phosphorylation of choline to produce phosphocholine, in human colon cancer and adenoma. Both choline kinase activity and phosphocholine levels were increased in colon cancer and adenoma tissue. The activation of choline kinase and the increased levels of choline kinase alpha were partly responsible for the elevated phosphocholine levels. This study suggests that choline kinase might play a role in growth promotion or signal transduction in carcinogenesis. PMID- 10363581 TI - Inhibition of experimental metastasis of human fibrosarcoma cells by anti recombinant 37-kDa laminin binding protein antibody. AB - The laminin binding protein of 37 kDa (37LBP) is regarded as a precursor protein of the high-affinity 67-kDa laminin receptor (67LR). Expression of 67LR/37LBP is well correlated with biological aggressiveness of cancer, particularly with invasive and metastatic potential. To investigate in detail the role of 37LBP in cancer cells, we synthesized recombinant 37LBP (r37LBP) as a fusion protein and generated an IgG-type polyclonal antibody P4G against r37LBP. Western blot analysis with P4G showed a single band of 67LR under both nonreducing and reducing conditions using cell extract of human fibrosarcoma cells HT1080. It was shown that P4G inhibited cell attachment to immobilized laminin in a dose dependent manner. Further, the intravenous injection of HT1080 cells pretreated with P4G, compared with that of cells pretreated with normal rabbit serum, resulted in a reduced number of experimental metastases (3.3+/-5.1 vs. 58.0+/ 38.0 nodules per mouse, respectively) (P<0.005). These results suggest that P4G inhibits the colonization and growth of HT1080 cells in the lungs of mice, and that the blocking of r37LBP with the specific antibody P4G may offer a potential strategy for preventing cancer metastasis. PMID- 10363582 TI - p53 status predicts the efficacy of postoperative oral administration of tegafur for completely resected non-small cell lung cancer. AB - Although postoperative adjuvant therapy for non-small cell lung cancer (NSCLC) had not been reported to be effective, it has been reported recently that oral administration of tegafur (1-[2-tetrahydrofuryl]-5-fluorouracil, FT) may improve the postoperative prognosis. In the present paper, to examine whether p53 status affects the efficacy of FT as postoperative adjuvant chemotherapy for NSCLC, a total of 236 consecutive patients with completely resected pathologic stage I IIIa NSCLC were retrospectively reviewed. p53 status was determined by immunohistochemical staining. For all patients, the 5-year survival rate of patients with FT administration (FT group) was 78.1%, being significantly higher than that (69.1%) of patients without FT administration (control group) (P=0.046). For patients without immunohistochemical evidence of p53 overexpression, the 5-year survival rate in the FT group was 87.1%, being significantly higher than that (74.0%) in the control group (P=0.036). This demonstrates an improvement of postoperative prognosis by FT administration. On the other hand, for patients with p53 overexpression, there was no significant difference in the postoperative prognosis between the FT group and the control group (5-year survival rate 63.2% and 60.1%, respectively; P=0.514), demonstrating that FT administration was not effective for these patients. In conclusion, p53 status may be useful for predicting the efficacy of postoperative adjuvant chemotherapy using FT. A prospective randomized study stratified by p53 status is needed to clarify the effect of postoperative FT administration. PMID- 10363583 TI - Differences in substrate specificity among glutathione conjugates (GS-X) pump family members: comparison between multidrug resistance-associated protein and a novel transporter expressed on a cisplatin-resistant cell line (KCP-4). AB - The substrate specificity of primary active transporters expressed on two kinds of human epidermoid KB-3-1 derived cell lines, C-A500 and KCP-4, was examined; the former expresses multidrug resistance-associated protein (MRP1), whereas the latter is resistant to cis-diamminedichloroplatinum (II) (cisplatin). Northern blot analysis indicated that neither P-glycoprotein, MRP1, MRP2 (canalicular multispecific organic anion transporter; cMOAT) nor MRP3 was overexpressed on KCP 4. Membrane vesicles isolated from C-A500 and KCP-4, but not from KB-3-1, exhibited the ATP-dependent uptake of glutathione conjugates (GS-X) such as leukotriene C4 and 2,4-dinitrophenyl-S-glutathione (DNP-SG), indicating the presence of GS-X pumps on these cells. The uptake of these GS-X by membrane vesicles from C-A500 was approximately twice that in the case of KCP-4. Kinetic analysis indicated that the Km and Vmax values for DNP-SG uptake were 2.56 and 1.43 microM, and 570 and 160 pmol/min/mg protein for C-A500 and KCP-4, respectively. In marked contrast, significant ATP-dependent uptake of glutathione platinum complex was observed only in membrane vesicles from KCP-4, but not those from KB-3-1 and C-A500. The transport properties of estradiol-17beta-D glucuronide (E(2)17betaG) were also different between the two cell lines. This was reflected in the findings that the ATP-dependent uptake of this conjugated metabolite in membrane vesicles from C-A500 (Km=2.33 microM, Vmax=34 pmol/min/mg protein) was much more extensive than that in the case of KCP-4 (Km=5.5 microM, Vmax=35 pmol/min/mg protein), and that comparable uptake was observed between KCP 4 and KB-3-1. Overall, a clear difference in substrate specificity among GS-X pump family members expressed on resistant tumor cells was demonstrated. PMID- 10363584 TI - Therapeutic effect of 1 M tegafur-0.4 M 5-chloro-2,4-dihydroxypyridine-1 M potassium oxonate (S-1) on liver metastasis of xenotransplanted human colon carcinoma. AB - S-1 [1 M tegafur (FT)-0.4 M 5-chloro-2,4-dihydroxypyridine (CDHP)-1 M potassium oxonate (Oxo)], was developed as a new oral antineoplastic agent based on biochemical modulation of fluorouracil (5-FU) by CDHP and Oxo. The therapeutic effect of S-1 on human colon cancer xenografts (TK-13) with high metastatic potential to the liver was evaluated. Small pieces of TK-13 were sutured into the cecal wall of 52 nude mice, and the animals were randomly divided into 3 groups [control (n=17), UFT (combination of 1 M FT and 4 M uracil) (n=18) and S-1 (n=17)]. S-1 or UFT was administered orally at an equitoxic dose (S-1, 7.5 mg/kg; UFT, 17.5 mg/kg as FT) for 37 consecutive days beginning 10 days after the transplantation. S-1 showed higher tumor growth inhibition than UFT (P<0.05) and also showed a significant anti-metastatic effect on liver metastasis, while UFT did not. Liver metastasis developed in only 2 of the 17 mice (12%) in the S-1 group, whereas it developed in 9 of the 17 (53%) and 7 of the 18 (39%) in the control and UFT group, respectively. Analysis of AUC (area under the curve) revealed that S-1 yielded higher 5-FU levels in both tumor tissue (1.6 times) and plasma (2.5 times) than UFT. These results suggest that S-1 will show a higher clinical therapeutic effect against human colorectal cancer than UFT. PMID- 10363585 TI - Enhancement of the anti-tumor effect of 5'-deoxy-5-fluorouridine by transfection of thymidine phosphorylase gene into human colon cancer cells. AB - Thymidine phosphorylase (dThdPase) is an enzyme that converts 5'-deoxy-5 fluorouridine (5'DFUR) to the toxic substance 5-fluorouracil (5-FU); it is also known to be a platelet-derived endothelial cell growth factor. In order to investigate the feasibility of suicide gene therapy against colorectal cancer by means of the combination of 5'DFUR and the converting enzyme dThdPase, we transfected the dThdPase gene into the human colon cancer cell line SW480 and analyzed the growth pattern as well as the sensitivity to 5-FU or 5'DFUR of the dThdPase-transfected cells. The 50% inhibition (IC50) values of 5-FU against the SW480 parental cells, control vector-transfected cells SW480/V1, and dThdPase transfected cells SW480/dThdPase were approximately 4.9, 6.3, and 2.9 microM, respectively. The IC50 of SW480/dThdPase was lower than that of SW480 or SW480/V1, although the differences were not statistically significant. The IC50 values of 5'DFUR for SW480, SW480/V1, and SW480/dThdPase were approximately 300, 330, and 3.2 microM, respectively. The sensitivity to 5'DFUR of SW480/dThdPase was increased by about 100-fold compared with that of SW480 or SW480/V1. With only 10% transfection efficacy, a high enough sensitivity to 5'DFUR was obtained to suppress the cell growth, indicating that a strong bystander effect was induced by this system. The in vivo growth of the s.c. transplanted SW480/dThdPase tumor in nude mice was significantly suppressed by i.p. injection of 5'DFUR compared with that in control mice that received phosphate-buffered saline (PBS) treatment. These results suggest that gene therapy using the combination of 5'DFUR and the dThdPase gene may be a useful approach for treatment of colon cancer. PMID- 10363586 TI - Ex vivo delivery of suicide genes into melanoma cells using epidermal growth factor receptor-specific Fab immunogene. AB - The Fab fragment of monoclonal antibody B4G7 against human epidermal growth factor (EGF) receptor was conjugated with cationic poly-L-lysine and the resulting conjugate was further complexed with reporter genes or therapeutic genes. This Fab/DNA complex was designated as "Fab immunogene." The Fab immunogene transfer in vitro was mediated through the EGF receptors in two melanoma cell lines. The frequency of cells expressing beta-galactosidase (beta Gal) reporter gene was approximately 1%. The induction of suicide effects after Fab immunogene transfer of herpes simplex virus thymidine kinase (TK) or Escherichia coli cytosine deaminase (CD) gene was quite remarkable, and the growth of melanoma cells was inhibited for over 7 days in the presence of ganciclovir (GCV) or 5-fluorocytosine (5-FC). Similarly, when melanoma cells treated in vitro with the Fab immunogene carrying TK or CD were transplanted into the back of nude mouse, subsequent systemic administration of GCV or 5-FC effectively suppressed the growth of tumors, indicating the occurrence of in vivo suicide effects. PMID- 10363587 TI - Frequent expression of midkine gene in esophageal cancer suggests a potential usage of its promoter for suicide gene therapy. AB - We have examined the expression of midkine (MK), a neurotrophic factor with heparin-binding activity, in human esophageal cancer cells. Seven esophageal cell lines tested expressed the transcript and 8 out of 14 human esophageal tumor specimens were positively stained with anti-MK antibody, while surrounding normal esophageal tissues in these specimens were not stained. The 5'-flanking, 2.3 kb genomic region of the MK gene was shown to drive the transcription of a reporter gene in the esophageal cell lines in a cis acting manner. Forced expression in esophageal cancer cells of herpes simplex virus-thymidine kinase gene mediated by the flanking region of the MK gene conferred sensitivity to a prodrug, ganciclovir. The 5'-upstream region of the MK gene thus possesses putative promoter activity which can be used for suicide gene-based gene therapy for esophageal cancer. PMID- 10363588 TI - Efficient production of adeno-associated virus vectors using split-type helper plasmids. AB - Adeno-associated virus (AAV) vectors are potentially useful vehicles for the delivery of therapeutic genes into human cells. To determine the optimal expression pattern of AAV proteins (Rep78, Rep68, Rep52, Rep40, and Cap proteins) for packaging the recombinant AAV genome, helper plasmids were split into two portions. In this study, two sets of split-type helper plasmids were prepared; i.e., 1) a Rep expression plasmid (pRep) and Cap expression plasmid (pCap), and 2) a large Rep expression plasmid (pR78/68) and small Rep plus Cap expression plasmid (pR52/40Cap). When AAV vectors were produced using these sets of split type helper plasmids at various ratios, the optimal ratio of (large) Rep expression plasmid and Cap expression plasmid was 1 to 9 for both sets. More importantly, the titers were comparable to or even higher than that of a conventional helper plasmid (pIM45) (4.9+/-2.1x10(11) vector particles/10 cm dish for pRep and pCap; 2.9+/-1.6x10(11) vector particles/10 cm dish for pR78/68 and pR52/40Cap; and 1.8+/-0.16x10(11) particles/10 cm dish for pIM45). Western analysis of AAV proteins suggests that the expression of a relatively small amount of large Rep and a large amount of Cap is important for optimal vector production. The present study shows that the AAV helper plasmid can be split without losing the ability to package the recombinant AAV genome, and provides us with valuable basic information for the development of efficient AAV packaging cell lines. PMID- 10363589 TI - Kidney preservation in the next millenium. AB - For the past decades, severe hypothermia has represented the foundation of organ preservation in clinical transplantation. Beneficial as hypothermia has proven to be in preserving grafts from heart-beating donors, hypothermia does not seem to provide the window necessary for the prospective evaluation of organ function. With the increasing use of non-heart-beating donors, it is logical to propose that if organs are to be evaluated prospectively, it will be necessary to preserve them at warmer temperatures. Since both glomerular and tubular functions are inhibited at temperatures below 18 degrees C, such a goal will necessitate organ preservation at a temperature above 20 degrees C. The principle of preservation at warmer temperatures is not new, but with future developments and approaches, successful realization appears within reach. In this overview, a brief history of previous attempts at warm preservation, in the context of the current status of kidney preservation, is presented. Future developments and approaches, with the potential for prospective testing of the function and enhanced resistance to ischemic damage, will be discussed. PMID- 10363590 TI - Tacrolimus versus cyclosporin A: a comparative study on rat renal allograft survival. AB - Although tacrolimus has been studied in a wide variety of experimental animal models, we are the first group to systematically study the effect of tacrolimus on rat renal allograft survival, as primary therapy and as anti-rejection therapy, in comparison with cyclosporin A (CyA). Renal grafts were transplanted from BN to LEW rats. Tacrolimus and CyA were administrated orally from day 0 for 50 days as primary therapy after grafting. Allografts were rejected after a median survival time (MST) of 8 days. Both tacrolimus und CyA significantly prolonged renal allograft survival, in a dose-dependent manner compared with the allograft controls. The most effective dose was 3.2 mg/kg, per day for tacrolimus, and 10 mg/kg per day for CyA. There was no significant difference in renal function between the group treated with the most effective dose of tacrolimus and the CyA-treated group. The percentage of detectable serum IL-2 level was 45% in the allograft control group, but was undetectable in groups treated with the most effective dose of tacrolimus or CyA at days 3 and 6 after grafting. On the other hand, no side effects were noted in recipient rats by daily inspection, body weight change, and histological studies, although minimal tubular vacuolation was encountered in the group treated with CyA 32 mg/kg per day. In addition, the most effective doses of tacrolimus and CyA were studied as anti-rejection therapy. All of the 5 recipients treated with tacrolimus from days 2-14, and 3 of the 5 treated from days 4-16 after grafting, survived for more than 50 days. However, the MST was 19 days for recipients treated with CyA from days 2-14, and 13 days for those treated from days 4-16 after grafting. In summary, tacrolimus as primary therapy induced rat renal allograft survival with renal function and side effects comparable with those of CyA. Interestingly, when both agents were used as anti-rejection therapy, tacrolimus, but not CyA, could significantly overcome ongoing renal allograft rejection in the rat. PMID- 10363592 TI - Cytoprotective effect of trimetazidine on 60 minutes of intestinal ischemia reperfusion injury in rats. AB - Trimetazidine (TMZ), a potent antioxidant agent, has been used to protect the myocardium, liver and kidney from ischemia reperfusion (IR) injury. We investigated the effect of TMZ, a cellular anti-ischemic agent and a free radical scavenger, on 60 min of warm intestinal IR injury in rats. Sprague-Dawley rats were divided into three groups: a sham-operated group (no IR injury, n = 8), an ischemic control group (control, n = 8), and a TMZ-treated group (3 mg/kg, n = 8). Malondialdehyde (MDA) levels, myeloperoxidase (MPO) activity, and mucosal damage were investigated after 120 min of reperfusion. MDA levels and MPO activity were more elevated and histopathological damage more severe in the control group than in the sham group (P < 0.05). MDA levels and MPO activity were lower and there was less histopathological damage in the TMZ group than in the control group (P < 0.05). Accumulation of lipid peroxidation products and neutrophils in mucosal tissues were significantly inhibited by TMZ treatment. We conclude that pretreatment of rats with TMZ before intestinal ischemia attenuates but does not prevent, histological damage. PMID- 10363591 TI - Plasma proteolytic activity in liver transplant rejection. AB - In this study, we evaluated the role of proteolytic enzymes belonging to the coagulation, fibrinolytic, and plasma contact systems in the early postoperative phase after orthotopic liver transplantation (OLT). Twenty-nine patients were studied at the time of OLT and during the first 2 postoperative weeks. Blood samples were collected daily after OLT and analyzed for kallikrein-like activity (KK), functional kallikrein inhibition (KKI), plasmin-like activity (PL), and alpha2-antiplasmin (AP). In addition, prekallikrein (PKK), prothrombin (PTH), antithrombin III (AT III), plasminogen (PLG), prothrombin/antithrombin III complexes (TAT), prothrombin fragment 1 + 2 (F1 + 2), and plasmin/alpha2 antiplasmin complexes (PAP) were measured. Nineteen patients experienced biopsy verified acute rejections (AR) and ten patients had uneventful courses and served as controls. Plasma analyses showed that the contact, coagulation, and fibrinolytic systems were activated during OLT. Following OLT, continuous thrombin and plasmin generation was observed, and these effects were more pronounced in the group having an uneventful course than in patients with AR. Factors that could possibly affect plasma proteolytic activity, such as blood product usage during and after OLT and cold ischemia time of the liver graft, did not differ between the groups, nor did the routine liver function tests, alanine aminotransferase (ALT) and aspartate aminotransferase (AST). PMID- 10363593 TI - Effects of lung preservation solutions on PMN activation in vitro. AB - Polymorphonuclear leukocyte (PMN) activation and PMN-endothelial cell interactions may cause graft failure due to ischemia-reperfusion injury after lung transplantation. We investigated the effects of Euro-Collins solution (EC), low-potassium dextran solution (LPD), and EC plus pentoxifylline (EC-PTXF) on adhesion molecule (CD11b/CD18 and L-selectin) expression, chemotaxis, and oxidative burst of PMN. PMN from healthy human volunteers were incubated with EC, LPD, and EC-PTXF, and, in controls, without preservation solution. LPD exerted a suppressive effect on PMN chemotaxis as compared to EC (P < 0.05), but had no attenuating effect on the increase of CD11b/CD18, the shedding of L-selectin, and intracellular oxidant generation. EC-PTXF attenuated the expression of CD11b/CD18 and the oxidative burst as compared to EC alone (P < 0.05). These effects of LPD and PTXF on PMN function may contribute to successful organ preservation in transplantation. PMID- 10363594 TI - A comparison of pediatric and adult kidney donors for adult recipients. AB - The high demand for organs for transplantation has made it necessary to consider using even the oldest and youngest of potential donors in order to increase the organ supply. In this retrospective study, the outcome of kidney transplantation using cadaveric pediatric donors was compared with that of an adult control series. Graft procurement took place in two regions of Italy (Emilia-Romagna and Piemonte) over an 11-year period. A group of pediatric donors (< 15 years old, n = 30) was compared with an adult donor group (n = 67). All recipients were adults who received cyclosporin as immunosuppression. Actuarial patient and graft survival rates did not differ significantly between the two groups (patient survival 96% and 96% for pediatric donors versus 98% and 92% for adult donors at 1 and 5 years post-transplantation; graft survival 76% and 68% for pediatric donors versus 88% and 74% for adult donors 1 and 5 y post-transplantation). Complications were also evaluated, but no difference was found (the only exception being the creatinine level in the 5th year). Renal transplantation with cadaveric donors starting at 4 years of age gave results comparable to kidneys coming from adults. These data show that cadaveric pediatric donor kidneys may be used in adult recipients with good results. The ethical implications of the subject are extensively reviewed. PMID- 10363595 TI - Contribution of color and power Doppler sonography to the differential diagnosis of acute and chronic rejection, and tacrolimus nephrotoxicity in renal allografts. AB - The aim of the present study was to differentiate acute rejection, chronic rejection, and tacrolimus nephrotoxicity with color and power Doppler imaging of renal transplants. One hundred examinations were obtained from 45 patients. Pulsatility and resistive indices were calculated from color Doppler images. The grade of renal vascularization was quantified using computer-assisted pixel analysis in a rectangular region-of-interest. The percentage of vessel-covered renal parenchyma (POV) was calculated using a histogram that discriminated renal vessels from renal parenchyma via power Doppler images. Furthermore, the distance from the most peripherally located vessels to the renal capsule (PVD) was measured. A reduced POV < or = 55% proved to be the best discriminator when chronic rejection was suspected (sensitivity 79%, specificity 87%). Tacrolimus nephrotoxicity showed not only a moderate elevation of the Doppler signal but also an increased PVD > or = 3.9 mm and a normal POV. We conclude that the evaluation of renal vessels by power Doppler images improves diagnostic accuracy for patients with renal allografts. PMID- 10363597 TI - Impact of local donor and regionalization on a German transplantation center. UNI NRW. Universities of North Rhine Westfalia. AB - Optimal allocation of donor organs is an ongoing matter of debate. We report on the impact of the foundation of UNI NRW, a close transplant collaboration of seven university centers with the intention of improving donor organ allocation, on the heart transplant program in Munster. All donor organs retrieved were offered first to the patients within this region before going into the Eurotransplant (ET) Foundation pool. The heart transplant program data were prospectively (for 1997) and retrospectively (for 1996) analyzed with regard to donor organ availability and allocation. There was a slight decrease in the number of donor hearts offered and accepted within the UNI NRW region in 1997 as compared to in 1996. However, due to the significantly lower organ export rate, the number of heart transplantations performed in UNI NRW rose from 47 to 72 procedures. In Munster, only six donor organs (16%) were procured from outside UNI NRW in 1997, and these were, in part, due to special urgency requests. In conclusion, the institutionalization of UNI NRW within the framework of ET offers more flexibility, decreases total ischemic time, and may help to lower costs. PMID- 10363596 TI - Renal graft survival in native and non-native European recipients. AB - Most studies on the influence of recipient race on kidney transplant survival have been performed in the United States. Generally, they show a lower survival in African-Americans than in Caucasians. Since Rotterdam has gradually become a multi-ethnic society, we were able to study the effect of origin on kidney survival. We restricted our study to recipients of a primary cadaveric kidney graft between July 1983 and July 1997 who received cyclosporin as primary immunosuppression. Patients were divided into two main groups according to origin: European (n = 399) and non-European (n = 110). No statistical differences were found for mean donor age, sex distribution, or the total number of HLA-A and DR mismatches. Non-Europeans had significantly more mismatches on their HLA-B locus (P = 0.01) and recipient age was lower (P = 0.003). The reason non Europeans had lost their native kidneys was more often hypertension and less often congenital or hereditary diseases compared to Europeans. The causes of death and of transplant failure did not differ. A multivariate Cox proportional hazards analysis did not show European or non-European origin to be an independent predictor of graft survival (two categories, P = 0.25). The variable origin in five categories did show an independent influence on graft survival, with Arab en African recipients running higher risks than European and Asian recipients. We conclude that, in our center, the prognosis after kidney transplantation is comparable for Europeans and non-Europeans; however, in the subcategories, Arab and African recipients have a worse prognosis. PMID- 10363598 TI - In vivo migration of lymphocytes in chronically rejecting rat kidney allografts. AB - Histological analyses have identified lymphocytes and macrophages as the predominant leukocyte populations that infiltrate organs undergoing chronic rejection. In order to define the time frame of this infiltration, we investigated the in vivo migration pattern of lymphocytes in a well-established rat model of chronic kidney allograft rejection. F344 kidneys were orthotopically transplanted into bilaterally nephrectomized Lewis rats. Recipients were treated with cyclosporin A (1.5 mg/kg/per day) for the first 10 days. After anti-CD18 or vehicle pretreatment, peripheral blood lymphocytes obtained from naive Lewis rats and labeled with 3H-uridine were injected into transplanted rats 12 and 16 weeks after transplantation. Organs were harvested 4, 8, and 12 h thereafter. After 12 weeks, proteinuria developed, accompanied by all signs of chronic rejection including glomerular sclerosis. Labeled lymphocytes rapidly infiltrated grafted kidneys 4 h after injection. Even more lymphocytes had accumulated in the grafts 12 h after injection. After 16 weeks, few lymphocytes had emigrated into the graft at 4 h, while infiltration was most pronounced by 12 h. Pretreatment with anti-CD18 inhibited the influx of lymphocytes. There was no difference between the patterns of lymphocytes derived from naive and transplanted rats. Our results emphasize the importance of endothelial cells in chronically rejecting kidneys for the control of leukocyte influx. Beta2-integrins may play a central role in determining the transendothelial migration during this process. PMID- 10363599 TI - "En bloc" paediatric renal donors into adult recipients -- the Newcastle technique. AB - Whilst debate still continues about the best use of kidneys from small donors, the techniques used have been varied because of the high vascular thromboses rates and ureteric leak rates. The method described here employs a vessel transposition as described by two German series, but it is combined with an extraperitoneal approach. It is now the method of choice in our unit for such en bloc transplants. PMID- 10363600 TI - FDG PET diagnosis of septic kidney in a renal transplant patient. PMID- 10363601 TI - Kidney graft dysfunction after drug interaction between miocamycin and cyclosporin. PMID- 10363602 TI - Expansion of donor pool: lack of function predictors. PMID- 10363604 TI - Chest re-exploration for complications after lung surgery. AB - BACKGROUND: Review of the most recent chest re-explorations for lung surgery complications may show methods by which risks can effectively be reduced. METHODS: The data on rethoracotomies following lung surgery over the past 14 years in our department were retrospectively reviewed. The indication, the type of operation, the outcome, and various factors influencing the postoperative mortality were analyzed. From 1983 to 1996, 1960 patients underwent primary thoracotomies for various lung diseases. Among these, 73 (3.7%) patients required re-exploration for various postoperative complications. RESULTS: Mean age was 56.8 years (15-80 years). There were 66 (90.4%) men and 7 (9.6%) women. The most common indication for rethoracotomy was hemorrhage in 38 (52%) patients. The source of bleeding was a mediastinal and/or bronchial blood vessel in 8 patients and an intercostal blood vessel in 6 patients. Six patients had to be reoperated because of hemorrhage from a major artery of the hilus. In 14 cases the postoperative hemothorax occurred without evident surgical origin. Further indications for rethoracotomy were bronchopleural fistula (BPF) in 13 (17.8%) patients, and persistent parenchymal leak in 8 (10.9%) patients. There were 8 additional causes distributed among the remaining 14 (19.3%) patients. The overall mortality rate was 17.8% (13/73), with the highest (38.4%) among BPF patients. CONCLUSIONS: Postoperative complications following lung surgery which require rethoracotomy are rare. The most common complication is postoperative bleeding. This is followed by bronchial stump insufficiency which is associated with the highest mortality and morbidity. Our experience shows that the need for re-exploration can hardly be reduced but the indication for re-exploration should be established as early as possible to avoid late complications. PMID- 10363603 TI - The effect of various doses of lazaroid U74389G on lung ischemia reperfusion injury. AB - BACKGROUND: We investigated the effect of lazaroid U74389G, a potent inhibitor of lipid peroxidation, on ischemia-reperfusion injury at three different doses in the rat orthotopic left lung transplantation model. METHODS: Four groups of reperfused lungs were studied. In group I (control) donor lungs were transplanted after 12 hours of preservation in University of Wisconsin (UW) solution at 4 degrees C. In groups II, III, and IV, lazaroid was used as an additive to UW solution at concentrations of 30 micromol/L, 50 micromol/L, and 100 micromol/L, respectively, and was also administered intravenously to the recipients 30 minutes before reperfusion after 12 hours of storage in the UW solution at 4 degrees C. RESULTS: After 1 hour of reperfusion, gas exchange function (p < 0.01), tissue lipid peroxide levels (p < 0.05) and histologic damage in reperfused lung allografts (p< 0.05) were significantly improved in lazaroid treated group IV compared with control group I. CONCLUSIONS: These findings suggest that lazaroid U74389G ameliorates ischemia-reperfusion injury in rat lung transplants by inhibiting lipid peroxidation. PMID- 10363605 TI - Minimally invasive treatment of thoracic empyema. AB - BACKGROUND: The present study was undertaken to assess the efficacy of serial thoracocentesis and saline irrigation for the treatment of pleural empyema, for post-pneumonia versus other causes. METHODS: Included were 42 consecutive patients with complicated pleural effusion (n=14) or frank pus (n=28) at diagnostic thoracocentesis, of mean age 57.5 +/- 23.7 years. Pneumonia was the probable cause in 29 patients, other causes (principally thoracic surgery) in 13. In addition to antibiotics in all patients, ultrasonography-guided serial suction thoracocentesis with saline irrigation was used as therapy of first choice in most patients: 28/29 post-pneumonia and 9/13 non-pneumonic empyema. Exceptions were mainly on the grounds of preceding thoracic surgery. RESULTS: Success rate was 86% in the post-pneumonia group, with no crossovers to more invasive therapy. Mortality was 14%, none empyema-related. Treatment was less successful in the non pneumonia group at 69%, with a 56% crossover rate from thoracocentesis due to therapy failure. Mortality was 23 %. CONCLUSIONS: Results indicate that timely, minimally invasive therapy is a feasible modality for the management of post pneumonia thoracic empyema. Serial thoracocentesis was less suitable for the treatment of non-pneumonic empyema, however, particularly if it was a surgical complication. More invasive strategies seem preferable in such cases. PMID- 10363606 TI - Postoperative death should be followed by autopsy - an analysis of the autopsy findings of the years 1990 and 1991 in a heart surgery center. AB - BACKGROUND: In this retrospective analysis of all autopsies performed in 1990 and 1991 on cardiac surgery patients who died before discharge from our university hospital, we intended to test the use of postmortem examination among cardiac surgery patients. METHODS: Perioperative data of all patients who underwent autopsy because of postoperative death during this time period were collected using a retrospective analysis of hospital and autopsy records. RESULTS: In 1990 and 1991 a total of 2407 patients underwent cardiac surgery with extracorporeal circulation at our institution. The in-hospital mortality was 2.9% (n=36) in 1990 and 3.3% (n=40) in 1991. For most of all patients who died during the postoperative course, we found a highly symptomatic cardiac disease and significant co-morbid conditions. The autopsy rate was 46.1% (35/76 patients). Cardiac failure with shock symptoms was the leading course of death (68.6%). In 22.9% of these patients (n=8) the autopsy provided information which had not been clinically recognized (e.g. myocardial infarction, thrombosis of bypass grafts, pneumonia) but might have altered the postoperative therapy if it had been. CONCLUSIONS: Despite the well-known trend of decreasing autopsy rates in the western world we believe that the postmortem examination is a most valuable diagnostic tool in the setting of a university cardiac surgery unit. Our results confirm the importance of autopsies for all patients who die after the operation, because a significant part of autopsies reveals major discrepancies between clinical and postmortem diagnoses. In an effort to maintain a high quality of treatment and education the autopsy rate of a hospital is of utmost interest. PMID- 10363607 TI - Action of aprotinin in myocardial ischemia - an investigation using a plasma-free model. AB - BACKGROUND: The protease inhibitor aprotinin has been reported to have an anti ischemic effect on left-ventricular myocardium in patients undergoing cardiopulmonary bypass operation. To examine the anti-ischemic properties beside its antifibrinolytic and inhibitory action on the kallikrein-bradykinin system, we investigated this substance in buffer-perfused rat hearts. METHODS: 24 isolated isovolumically contracting rat hearts received a 10-minute infusion of either 10000 units aprotinin or pure saline followed by 30 minutes of no-flow global ischemia and 45 minutes of reperfusion. Hemodynamics, high-energy phosphates, and troponin T as molecular marker of cardiac injury were studied. RESULTS: During 15 minutes of reperfusion steady state function was identical in both groups, with a recovery of the developed left-ventricular pressure to 81.9+/ 1.5% after protease inhibition and 83.0+/-2.6% in the controls. Coronary flow, myocardial oxygen consumption, and contractile reserve after maximum Ca++ stimulation were also identical. High-energy phosphates were comparably reduced in both groups (adenine nucleotides: 3.1+/-0.3 micromol/g ww after aprotinin vs. controls 2.7+/-0.4 micromol/g ww and creatine phosphate: 6.5+/-0.9 micromol/g ww vs. controls 4.7+/-1.1 micromol/g ww). However, release of the cardiac specific marker troponin T was lower after ischemia at several measurements (p<0.05). The total release of troponin T was 44+/-10 ng in the aprotinin treated hearts vs. 90+/-17 ng in the postischemic control hearts (p<0.05). CONCLUSIONS: The findings demonstrate that aprotinin in a moderate dose is effective in reducing postischemic troponin release in a non-blood perfused system. Measurement of myocardial high-energy phosphates after aprotinin use was performed for the first time and indicates that not a reduction in severity of direct myocardial ischemic intensity but a beneficial action on processes causing release of troponin is the mode of action of this effect. PMID- 10363608 TI - Percutaneous transluminal septal ablation in hypertrophic obstructive cardiomyopathy. AB - BACKGROUND: Percutaneous transluminal septal myocardial ablation (PTSMA) by alcohol-induced occlusion of septal branches with resulting reduction of LV outflow-tract gradient (LVOTG) is a new treatment option in symptomatic patients with hypertrophic obstructive cardiomyopathy (HOCM). METHODS: In 1996 and 1997 we treated 114 symptomatic patients (56 female; age 53.3 +/- 15.6 years; 5 patients with prior myectomy and 5 with DDD pacer; most in NYHA class III. Five patients underwent re-PTSMA after failed first treatment. In the first 30 patients 1 to 3 septal branches were occluded by injection of 3.4 +/- 1.6 ml absolute alcohol via the central lumen after balloon occlusion of the proximal part of the septal branch. In the remaining patients myocardial contrast echocardiography was available, so that only one branch needed to be occluded. RESULTS: LVOTG reduction was achieved in 107 (94%) patients: at rest from 73.8 +/- 36.5 to 18.6 +/- 19.7 mmHg (p < 0.00001). Maximal CK rise was 647 +/- 330 U/L. Two (1.8%) patients died during hospital stay. Due to permanent trifascicular block 11 (9.6%) patients required a permanent pacemaker. At 3 months follow-up in 87 patients we observed no cardiac complications, a further LVOTG reduction in 61 % patients, an ongoing symptomatic improvement (NYHA I or II; p < 0.0001 vs. pre PTSMA), and significant reduction of the left posterior wall thickness. CONCLUSIONS: PTSMA of HOCM results in significant reduction of LVOTG. Careful monitoring during hospital stay is necessary because of the potential risks of the induced therapeutic infarction. Mid-term follow-up showed ongoing symptomatic improvement without cardiac complications. Remodeling after circumscribed septal infarction results in further LVOTG reduction in over 50% of the patients. PMID- 10363609 TI - The influence of different strategies on clinical outcome in patients undergoing total cavopulmonary connection. AB - BACKGROUND: We report on results of a prospective clinical trial designed to demonstrate the influence of various strategies in "Total Cavopulmonary Connection" (TCPC) for palliative therapy of patients with "single ventricle" physiology. METHODS: From 1989 to 1997, a total of 47 patients (mean age 4.8 +/- 3.6 years) underwent definitive TCPC at our unit. 31 patients (66%) underwent one stage TCPC, in 16 patients (34%) we performed a two-stage modified Fontan operation; 21 patients had central fenestration (4 mm). Inhalative NO therapy in the immediate postoperative period was adopted in 1993. RESULTS: Overall 5-year survival was 76.4%, after two-stage TCPC 87.5%, and 81.3% in patients undergoing fenestrated procedures. Two of three patients survived perioperative Fontan take down. We lost 11 patients (nine early and two late deaths): three patients died primarily because of neurologic dysfunction and eight patients because of cardiac failures. Under perioperative NO therapy there was no early death. After a mean follow-up of 35.9 +/- 23.3 months, 76% of all patients were in NYHA I and 21 % in NYHA I-II. 89.7% had sinus rhythm. 42% of our patients suffered from temporary pleuropericardial effusions. CONCLUSIONS: Definitive palliation with TCPC achieves acceptable clinical results. Two-stage repair, fenestration, and postoperative inhalative NO therapy - each have a positive influence on early and long-term survival. PMID- 10363610 TI - Surgical management of cardiovascular lesions caused by systemic inflammatory diseases. AB - BACKGROUND: The surgical treatment of cardiovascular disorders caused by inflammatory diseases presents many difficulties, including suture detachment and progression of vascular lesions. We here report the various surgical procedures used to treat these disorders and their long-term outcomes. METHODS: We operated on 14 patients: eight with Takayasu's disease, three with systemic lupus erythematosus, two with rheumatoid arthritis, and one with Behcet's disease. Patients were divided into three groups as follows; patients (n=7) requiring aortic valve replacement; patients (n = 4) requiring reconstruction of the coronary artery; and patients (n = 3) requiring aortic aneurysm repair. RESULTS: There were no early or late deaths in the postoperative follow-up period of 70 +/ 40 months, but there was one operative death. Three patients received postoperative steroids due to progression of the inflammation. However, there were no major complications such as valve detachment, pseudoaneurysmal formation, or occlusion of the bypass conduit. CONCLUSIONS: We conclude that it is crucial to reduce inflammation pre- and postoperatively, to reinforce the suture line, and to carefully select the operative procedures when treating cardiovascular disorders caused by inflammatory diseases. PMID- 10363611 TI - Reducing the post-pump syndrome by using heparin-coated circuits, steroids, or aprotinin. AB - BACKGROUND: Cardiopulmonary bypass (CPB) induces a systemic inflammatory response called 'post-pump syndrome'. As a part of a complex interaction between white cells and vascular endothelium, proinflammatory cytokines IL-6 and IL-8 are part of a phased immune response that is also balanced by anti-inflammatory cytokines such as IL-10. We compared the influence of heparin-coated circuits, steroids, and aprotinin on these cytokines, looking for ways to reduce the syndrome. METHODS: 40 patients with coronary artery disease (CAD) undergoing elective CABG were prospectively studied in four randomized groups of 10. Group A received prednisolone pre- and postoperatively (2 x 250 mg), group B received aprotinin perioperatively (6 Mio. KIU). In group C, heparin-coated circuits ('Bioline' by Jostra) were used and in group D no special measures were taken (controls). Plasma levels of cytokines were measured before and during CPB and until 12 h after surgery using an ELISA technique. RESULTS: In group A IL-6 was significantly (p<0.05) suppressed in contrast to the control group (A: peak at 4 h, 155 pg/ml vs. control: peak at 8 h, 565 pg/ml). IL-8 was also suppressed (A: peak at 30', 22 pg/ml vs. control: peak at 30', 55 pg/ml). IL-10 level changed first and was markedly upregulated in contrast to the control (A: peak at 30', 1600 pg/ml vs. control: peak at 30', 130 pg/ml; p<0.05). In group B (aprotinin) the cytokine release was similar to group A. Using heparin-coated circuits (group C) also led to a significant (p<0.05) IL-10 upregulation (C: peak at 2 h, 1380 pg/ml) and IL-6 suppression (C: peak at 4 h, 290 pg/ml). IL-8 was not influenced significantly. CONCLUSIONS: The results show a similar reduction of the inflammatory cytokine release (IL-6 and IL-8 as markers) using early steroid application and aprotinin in high dosage. Heparin coating reduces IL-6 and increases IL-10 release, whereas IL-8 is not affected. Further studies should investigate the effects of a combined application for reducing inflammatory cytokine release and the post-pump syndrome. PMID- 10363612 TI - Endoluminal grafting of infrarenal aortic aneurysms. AB - BACKGROUND: The department policy regarding therapy fo infrarenal aortic aneurysms is reviewed, based on the treatment results of a 12-months period. METHODS: From October 1996 to August 1997, 60 patients with infrarenal aortic aneurysms were admitted to our department. Of these 31 (52%) were found to be anatomically or pathomorphologically suitable for endovascular treatment, based on the premises that: 1. Whenever the anatomy is suitable and confirmed with CT or angiography, repair is by means of stent placement. 2. In emergencies and in cases where the anatomical relationships are unfavourable, patients undergo conventional open surgery. RESULTS: In all 31 patients treated endovascularly, stent placement was technically successful. Procedure-associated mortality was zero. The following stenting complications occurred: seven endoleaks, one thrombotic iliac occlusion, one femoral arterial dissection, two puncture-related inguinal hematomas. Elective open surgery was performed in the other 29 patients. One of these died from the effects of renal failure. CONCLUSIONS: This comparison shows that endovascular treatment of infrarenal aortic aneuryms is possible in a large proportion of patients and is not associated with an unfavourable rate of complications. Endovascular treatment can significantly reduce patients' postoperative hospitalization (three days) and time spent in intensive care. PMID- 10363613 TI - Thoracoscopic resection of intrathoracic vagus neurofibroma. AB - A thoracoscopic resection of mediastinal neurofibroma originating from the intrathoracic vagus nerve was successfully performed in an 18-year-old female. This report reviews the clinical and morphological features of intrathoracic vagus neurofibroma. Often neurofibroma has multiple lesions and/or multinodular shape. Thoracoscopic resection is a useful treatment for patients with intrathoracic neurofibroma, although careful observation is necessary when performing a radical resection. PMID- 10363614 TI - Acute respiratory insufficiency in an infant caused by a tracheogenic cyst. AB - The unusual case of acute respiratory insufficiency in an infant of 5 months caused by a large cyst of true tracheal origin is presented. PMID- 10363615 TI - Combined surgical approach for sarcoma lung metastasis with atrial involvement. AB - A 20-year-old patient, who had been treated for a femur sarcoma with pulmonary metastases 8 years before, arrived at our institution with a new metastatic hilar lung nodule. During the standard lobectomy procedure an unexpected atrial invasion by the tumor was discovered. Intraoperative transesophageal echocardiography (TEE) showed a big pediculated tumor in the atrium. Cardiopulmonary bypass (CPB) was required in order to safely resect the atrial wall with the tumor. The atrial defect was repaired with a pericardial patch. Postoperative course was uneventful. After 14 months, the patient is asymptomatic and free of disease. PMID- 10363616 TI - Left-ventricular inflow obstruction due to a dilated coronary sinus mimicking Cor Triatriatum. AB - Persistence of the left superior vena cava with drainage to the coronary sinus is a common congenital anomaly. We report an infant with such a malformation associated with marked enlargement of the coronary sinus, which produced partial supramitral obstruction and consequently impairment to the left-ventricular inflow. The patient pre-sented with cardiac failure in infancy and features mimicking cor triatriatum. Surgical relief of the supramitral obstruction resulted in immediate reversal of the pulmonary hypertension, with clinical improvement. This rare entity, only once previously reported, is an unusual cause of pulmonary hypertension in infancy. PMID- 10363617 TI - Minimally invasive extirpation of a left-ventricular myxoma. AB - As myxomas usually are benign neoplasms a minimally invasive technique would be an interesting alternative for their extirpation because the surgical trauma is reduced. In one male patient with a diagnosed left-ventricular myxoma minimally invasive surgery was carried out using the Port-Access method. 2D echocardiography, EBT, and MRI were performed preoperatively to obtain exact information about topography, calcifications, and malignity. Minimally invasive extirpation was successful and the mitral valve could be preserved. Histopathological examination revealed a cardiac myxoma extirpated in toto. At one-year follow-up there was no recurrence of the tumor. PMID- 10363618 TI - The disproportion of female and male surgeons in cardiothoracic surgery. AB - In ancient times, female medical practitioners and female surgeons were well known. With the introduction of medicine as an academic course and the ban on women studying, the medical career became virtually impossible for women. This condition changed with the general admission for women to colleges, in Germany in 1908. The current situation of women in surgery is presented here, with cardiovascular surgery as an example. Of 302 active cardiothoracic and cardiac surgeons 4.6% are female. According to an inquiry of all German heart institutes, there are 20 female senior registrars and 27 female surgeons as well as 162 female assistants. Most of the medical directors do have a positive opinion about female surgeons, but criticize that only a few women who apply for actually finish surgical training. One reason for this may be the greater difficulties for women to take care of a family and become a surgeon simultaneously. In this regard, an improvement in the position of female doctors is desirable. PMID- 10363619 TI - Importance of pharmacological and physicochemical properties for tolerance of antimuscarinic drugs in the treatment of detrusor instability and detrusor hyperreflexia--chances for improvement of therapy. AB - BACKGROUND: Antimuscarinic side-effects are relatively frequent problems in oral pharmacotherapy of detrusor instability and neurogenic dysfunction of the urinary bladder. Results of recent clinical trials demonstrate differences in tolerance between antimuscarinic drugs. It is the purpose of this paper to relate the available clinical data to the pharmacological and physicochemical properties of the different antimuscarinic drugs, in order to discuss the reasons for this enhanced tolerance and to make possible modes for improvement of antimuscarinic therapy plainly visible. METHODS: Therefore, we reviewed the available literature using among others the computerized library systems Medline (National Library of Medicine, Bethesda, Maryland, USA) and Embase (Excerpta Medica, Amsterdam, the Netherlands). Differences in tolerance of oral antimuscarinic drugs may result from muscarine-receptor selectivity, organ selectivity, and pharmacokinetic as well as physicochemical properties. While the roles of m-receptor and organ selectivity need more detailed clarification, influences of differences in bioavailability and physicochemical properties on the tolerance of antimuscarinic drugs are more sufficiently investigated. RESULTS: Generally, tolerance as well as efficacy of antimuscarinic drugs seem to be a complex result of a combination of various pharmacological properties distinguishing the individual substances. The enhancement of tolerance of propiverine hydrochloride, tolterodine tartrate and trospium chloride compared to oxybutynin chloride seems to be reached by different modes, from which the molecular structure -- propiverine and tolterodine are tertiary amines, trospium chloride possesses a quarternary ammonium structure -- may be of great importance. First investigations with alternative transdermal and intravesical application routes show interesting possibilities for further improvement of antimuscarinic therapy in urological indications. CONCLUSION: In conclusion, from pharmacological and clinical data it becomes obvious that there are significant differences between antimuscarinic drugs, which are of clinical relevance and include possible starting points for the development of new drugs and application forms. PMID- 10363620 TI - Pharmacokinetics and bioavailability of the flavonol quercetin in humans. AB - Flavonoids are plant polyphenolic compounds present in the daily diet. Latest epidemiological studies point to a crucial role of the flavonol quercetin in the prevention of cardiovascular diseases. It is assumed that this protective effect derives from the antioxidative capacity which quercetin shows in in vitro experiments. The antiproliferative and antimutagenic activities in vitro have made it a candidate for clinical trials in cancer therapy. Quercetin is also regarded as a putative active compound in various phytopharmaceuticals. However, in vivo data on the disposition, absorption, bioavailability, and metabolism of quercetin after intravenous and oral administration in humans are scarce and contradictory. The pharmacokinetic parameters following intravenous injection were determined in two studies. The elimination half-life was reported to be 2.4 h and 0.7 h, the volume of distribution at steady-state was 92.6 l and 6.2 l, and total body clearance was 34.6 lxh(-1) and 28.1 lxh(-1), respectively. Absorption after oral administration ranged from 0 to over 50% of the dose. These inconsistencies can partly be attributed to a lack of highly sensitive and specific assay methodology. The data available so far are insufficient to clarify whether or not quercetin can be held responsible for any protective or curative effect observed after oral intake. PMID- 10363621 TI - Phenazone as a marker of liver-metabolic function in patients with acute leukemia. AB - OBJECTIVE: The aim of this work was to study the influence of neoplastic disease, especially acute myeloblastic leukemia (AML), and its chemotherapy on the activity of hepatic microsomal enzymes by using phenazone, as a marker of oxidative drug metabolizing activity. METHODS: The observations were carried out in 21 patients with AML and in 53 healthy volunteers. The influence of disease on phenazone kinetics was studied before chemotherapy and the effect of anticancer drugs administration after the first cycle of chemotherapy. RESULTS: The mean phenazone half-life time was significantly shorter in patients with AML (8.79 (3.01) h) than in control group (11.08 (3.61) h) (p < 0.012). Treatment with anticancer drugs, especially with epirubicine, inhibited phenazone elimination. The mean phenazone half-life time was significantly longer (18.08 (8.80) h) and the mean metabolic clearance rate was significantly smaller (33.92 (15.40) ml/min) after chemotherapy in comparison with the initial value, before treatment (10.22 (2.90) h), (p < 0.01) (50.33 (20.29) ml/min) (p < 0.008). CONCLUSION: Our results lead to the conclusion that phenazone is an important index of hepatic metabolic capacity in patients with acute myeloblastic leukemia. The evaluation of its kinetics allowed to early recognition of the presence and the degree of drug oxidizing modification. Acceleration of phenazone elimination before treatment and its inhibition after chemotherapy, particulary epirubicine, may suggest that in patients with AML elimination of the other drugs metabolized by the pathway similar to phenazone also may be changed. It should be considered in individualization of their dosage regimen. PMID- 10363622 TI - Comparison of caudal tramadol vs bupivacaine for post-operative analgesia in children undergoing hypospadias surgery. AB - In a prospective double-blind study, 40 children scheduled for hypospadias repair were allocated randomly to receive either caudal tramadol (1 mg/kg) or 0.25% plain bupivacaine (0.5 ml/kg). Postoperative pain score, side-effects and oxygen saturation (SaO2) were recorded during 24-hour observation period. The results point toward a significantly lower pain scores with caudal bupivacaine in the immediate postoperative period, whereas caudal tramadol caused a significantly lower pain score in the late postoperative period. Total consumption of rescue analgesics was significantly higher in bupivacaine group as compared to tramadol group during the study period (p < 0.001). The incidence of side-effects such as vomiting was more frequent with caudal tramadol, but there was no detectable difference in SaO2. We conclude that caudal tramadol can safely be used for postoperative analgesia with a longer duration as compared to caudal bupivacaine. PMID- 10363623 TI - Topical netilmicin compared with tobramycin in the treatment of external ocular infection. AB - In a pilot double-blind, randomized, prospective controlled study the effectiveness and safety of 0.3% netilmicin ophthalmic solution were compared with those of 0.3% tobramycin in treating external bacterial ocular infections in 45 eligible patients. The treatment with both study medications resulted in a significant (p < 0.001, Wilcoxon test) reduction in the mean cumulative score of the signs and symptoms. However, no statistically significant differences were observed between the two groups. The clinical improvement rate was almost complete with either antibiotics. There was a statistically positive trend in the netilmicin group with regard to the microbiological improvement that was achieved in (87% of the netilmicin patients) compared with 77% of the tobramycin patients (77%). Antibiotic sensitivity revealed that 84% of the organisms isolated were sensitive to netilmicin whereas only 64% of them were sensitive to tobramycin. Only minor adverse events occurred in patients treated with either netilmicin or tobramycin. In conclusion, this study demonstrates that netilmicin is a promising new antibiotic for treating external ocular infections. PMID- 10363624 TI - Antacids have no influence on the pharmacokinetics of rabeprazole, a new proton pump inhibitor, in healthy volunteers. AB - AIM: This randomized, open-label, crossover study was conducted to evaluate the effect of a single dose of antacid, aluminum and magnesium hydroxide suspension, on pharmacokinetics of rabeprazole in healthy male volunteers. SUBJECTS AND METHODS: Twelve subjects were randomly divided into 6 groups of 2 subjects each, and received 20 mg of rabeprazole either without antacid, concomitantly with antacid, or one hour after administration of antacid in three experimental arms with washout period of one week. The concentrations of rabeprazole in plasma were determined by high performance liquid chromatography. RESULTS: Rabeprazole was well tolerated at the 20 mg dose level when given with or without antacid. The pharmacokinetic parameters, Cmax, t(max), AUC and t1/2, showed no statistically significant differences when rabeprazole was administered alone, concomitantly with antacid or one hour after antacid administration. CONCLUSION: No influence of single dose of antacid on pharmacokinetics of rabeprazole was observed. PMID- 10363625 TI - Pharmacokinetics of etoposide after intrathoracic instillation to lung cancer patients with pleural effusion. AB - OBJECTIVE: To examine etoposide (VP16) levels in serum and pleural effusion after intravenous infusion or intrathoracic instillation to lung cancer patients. METHODS: Four patients were administered VP16 by intrathoracic instillation and three patients were administered it intravenously. Serum, urine, and pleural effusion were collected and VP16 levels in the biological fluids were determined by HPLC. Pharmacokinetic parameters were calculated. RESULTS: VP16 distributed rapidly into pleural effusion after intravenous infusion. In two of three patients, VP16 levels in pleural effusion were maintained at constant levels more than 24 hours in spite of the decline in serum VP16 levels. After intrathoracic instillation, VP16 in pleural effusion reached high levels and eliminated slowly. Serum levels of VP16 were relatively low compared with those in pleural effusion. CONCLUSION: It was demonstrated that intrathoracic instillation of VP16 might be useful for managing malignant pleural effusion and reducing systemic side-effects by cutting down the dose. PMID- 10363626 TI - LDL receptor gene expression in human mesangial cells under the influence of calcium channel blockers. AB - BACKGROUND: Intracellular transport of lipid through the regulation of LDL receptor (LDLr) may be important in the progression of renal dysfunction. METHODS: We explored LDLr gene expression in human mesangial cell line (HMCL) under influence of calcium channel blockers using cell proliferation, LDL binding, Northern blot and LDLr promoter activity assay. RESULTS: Diltiazem and verapamil increased the expression of LDLr mRNA in a dose-dependent manner. Increased LDLr mRNA paralleled LDL binding. Nifedipine did not increase the expression of LDLr mRNA and LDL binding to HMCL at 1 - 100 micromol/l. The LDLr promoter activity assay showed that treatment with 100 micromol/ of diltiazem and verapamil increased LDLr promoter activity by 126.72 +/- 10.68%, and 166.41 +/- 11.41%, respectively, at 24 hours (control as 100%), while treatment with 100 micromol/l of nifedipine had an inhibitory effect on LDLr promoter activity. High concentration of LDL (250 microg/ml) inhibited promoter activity. Diltiazem or verapamil coincubated with LDL (250 microg/ml) could not override transcriptional inhibition by LDL. CCBs inhibited the proliferation of HMCL, therefore, CCBs induced LDLr expression did not depend on a proliferative response. Signal transduction pathway experiments showed that the calmodulin transduction pathway was involved in LDLr upregulation induced by diltiazem or verapamil. Additionally, tyrosine kinase and PKC pathways were involved in the induction of LDLr induced by verapamil. CONCLUSION: These studies show that diltiazem and verapamil increase LDLr gene transcription and expression which is independent of cell proliferation in HMCL. PMID- 10363627 TI - Neuropeptide Y Y1 receptor polymorphism as a prognostic predictor in Japanese patients with IgA nephropathy. AB - BACKGROUND: Neuropeptide Y exhibits a vasoconstricting action and regulates systemic blood pressure as well as noradrenalin. There are 5 types of NPY receptors, Y1 - Y5, which were introduced by pharmacological differences. Recently, a single point mutation in the first intron of the NPY Y1 receptor (NPYY1R) was reported. SUBJECTS AND METHODS: In this study, we investigated the relationship between NPYY1R gene polymorphism and clinical characteristics in patients with IgA nephropathy using polymerase chain reaction and restriction fragment length polymorphism analysis. RESULTS: Distribution of the NPYY1R genotypes which were defined as YY, Yy and yy genotypes, did not differ between 60 normal control subjects and 68 patients with IgA nephropathy (15 : 36 : 9 versus 21 : 40 : 7, respectively). In IgA nephropathy patients, the incidence of hypertension and the rate of urinary protein excretion were slightly higher in the non-YY genotype than in the YY genotype group (23% versus 5% and 1.1 +/- 1.2 versus 0.6 +/- 0.4 g/24 h, p = 0.09 and p = 0.05, respectively). The reciprocal of the serum creatinine level was estimated to determine the deterioration in renal function during follow-up after the renal biopsy. The level was lower in the non-YY genotype than in the YY genotype group (-0.002 +/- 0.064 vs 0.033 +/- 0.053/month, respectively, p < 0.01). Multiple regression analysis for the reciprocal of the serum creatinine level revealed that the NPYY1R genotype was an effective variable (p < 0.01). CONCLUSION: In conclusion, we propose that the NPYY1R gene polymorphism may be a novel prognostic predictor in patients with IgA nephropathy. PMID- 10363628 TI - Evidence for a link between glycoxidation and lipoperoxidation in patients with chronic renal failure. AB - AIM: The purpose of this study is to examine whether or not there is a relationship between glycoxidation and lipid peroxidation in patients with chronic renal failure. SUBJECTS AND METHODS: Dermal samples from 26 living or autopsied subjects were sequentially extracted with NaCl, pepsin, collagenase, and NaOH to obtain four fractions (salt-soluble fraction: SSF; pepsin-soluble fraction: PSF; collagenase-soluble fraction: CSF; and insoluble fraction: ISF). The glycoxidation product was measured by pentosidine-linked fluorescence (ex: 335/em: 385) and the levels of lipid peroxide, malondialdehyde (MDA), were assessed by determining the MDA-linked fluorescence (ex: 390/em: 460) which was further confirmed by HPLC. RESULTS: In patients undergoing hemodialysis, MDA linked fluorescence markedly increased in collagen-rich fractions, PSF, CSF, and ISF, while pentosidine-linked fluorescence increased in PSF and CSF, in comparison to the controls and the pre-dialysis patients with CRF. Interestingly, the increase in the lipid peroxides strongly correlated with the level of glycoxidation product in PSF, CSF, and ISF (p < 0.0001 in PSF, CSF; p < 0.01 in ISF). The HPLC data of MDA in the PSF was in good correlation with logistic levels of both MDA- (n = 9, r = 0.738, p = 0.023) and pentosidine-linked fluorescence (n = 9, r = 0.721, p = 0.028). In contrast, in SSF, the collagen poor fraction (collagen content: less than 3% of the total extracted collagen), the data showed a significant increase in the MDA-linked fluorescence only in the pre-dialysis patients with CRF, but not in the HD patients with no correlation with the glycoxidation products. CONCLUSION: These findings suggest that both the lipid peroxidation and glycoxidation increased in close relation to each other in the matrix collagen and thus demonstrate a synergetic contribution to the tissue damage observed in patients with CRF PMID- 10363629 TI - Hypertension is associated with hyperlipidemia, coronary heart disease and chronic graft failure in kidney transplant recipients. AB - BACKGROUND: Hypertension is a common concomitant condition in renal transplant recipients. There is accumulating evidence that this disorder is an important risk factor for chronic renal graft failure and other cardiovascular complications in these patients. SUBJECTS AND METHODS: The current retrospective study in 330 patients treated with cyclosporin or azathioprin covered 5 years and aimed to further characterize the interrelation between hypertension and renal graft failure. Furthermore, the association of hypertension with hyperlipidemia and the prevalence of coronary heart disease was evaluated. RESULTS: Altogether, before transplantation 182 patients were normotensive (no antihypertensive medication except diuretics) and 105 were hypertensive (blood pressure > 160/95 mmHg or patients requiring antihypertensive medication); for the remaining 43 patients no data were available. After transplantation the prevalence of hypertension in the cyclosporin group was 71, 76 and 70% after 1, 3 and 5 years, respectively. The respective numbers for the azathioprin group were 60, 59 and 58%. Hypertension was associated with graft dysfunction both in cyclosporin- and azathioprin-treated patients. Hyperlipidemia (cholesterol, triglycerides) was more severe in hypertensive than in normotensive patients. The prevalence for hypertension was higher in patients with coronary artery disease than in patients without the disease. CONCLUSION: The results further support the view that hypertension may be a risk factor for the development of chronic renal graft failure and coronary artery disease in this population. Furthermore, the association of hypertension with hyperlipidemia hints to an unfavorable accumulation of renal and cardiovascular risk factors in a large number of renal allograft recipients. PMID- 10363630 TI - Measurement of serum leptin in patients with chronic renal failure on hemodialysis. AB - BACKGROUND: Leptin, the product of the obese gene, is produced exclusively in fat cells. SUBJECTS, MATERIALS AND METHODS: To evaluate the clinical significance of measuring serum leptin in 56 patients with chronic renal failure on hemodialysis (HD), we measured leptin levels using radioimmunoassay in 34 normal volunteers and in 56 patients on HD. RESULTS: Normal serum leptin averaged 5.7 +/- 0.7 (mean +/- SEM) ng/ml, which correlated significantly (p < 0.001) with the body fat percentage as measured by bioelectrical impedance analysis. Serum leptin in HD patients ranged from 1.3 to 142 ng/ml. The mean serum leptin analyzed after the logarithmic conversion was 5.6 ng/ml, which was not significantly different from the normal control value, although the body fat percentage was significantly lower than normal volunteers. There was a significant (p < 0.01) positive correlation between body fat percentage and serum leptin in both normal controls and HD patients. The slope of the regression curve was steeper in HD patients than in normal controls. CONCLUSION: (1) serum leptin levels to body fat mass are significantly higher in HD patients than controls; (2) the variability is much wider in HD patients; and (3) a significant relation exists between percent body fat and log serum leptin, the relation being steeper in HD patients than in controls. PMID- 10363631 TI - Improved cardiovascular variables during acetate free biofiltration. AB - BACKGROUND AND AIM: Acetate free biofiltration (AFB) provides a well-tolerated and efficient renal replacement therapy. Replacement of most of the acetate by bicarbonate in standard hemodialysis has resulted in a decrease in intradialytic hypotensive episodes. This has been attributed to a decrease in the acetate induced impairment of myocardial contractility. The aim of the present study was to investigate whether the total absence of acetate in AFB would further enhance dialysis stability and improve cardiovascular status. PATIENTS AND METHODS: In a long-term, randomized trial we included 11 patients on AFB and 9 patients on bicarbonate hemodialysis (HD) for one year. Patients were matched for age, sex and urea reduction rate, but not for the presence of hypertension or cardiovascular history. During each dialysis session blood pressure was measured automatically and the presence of significant hypotension was recorded. Antihypertensive medication was registered every three months. Before and at the end of the study M-mode echocardiography was performed and left ventricular mass index (LVMi) was calculated. Every six months serum lipids were measured. RESULTS: At baseline, mean arterial pressure (MAP) before and after dialysis, the percentage of hypotensive dialyses, LVMi and serum lipids did not differ between AFB and HD. Pre-dialysis MAP decreased in AFB (from 112.5 to 107 mmHg) and increased in HD (from 101.7 to 105.3 mmHg; p = 0.01, HD versus AFB). Postdialysis MAP remained stable in both groups (AFB 91.6 mmHg at 0 months and 90.6 mmHg at 12 months, for HD respectively 83.9 and 86.5 mmHg, NS). The percentage of hypotensive dialyses did not differ significantly between the groups during the study. LVMi decreased in AFB from 195.4 to 162.1 gr/m2 and increased in HD patients from 153.8 to 182.5 gr/m2 (p = 0.03 HD versus AFB). The number of antihypertensive medications per patient did not differ between groups. Serum lipids remained unchanged during the trial. CONCLUSION: In conclusion, AFB provided better control of pre-dialysis MAP compared to HD, and stable postdialysis MAP. The percentage of dialysis sessions with hypotension did not differ. LVMi decreased significantly in AFB, but rose in HD. PMID- 10363632 TI - Acute eosinophilic interstitial nephritis and uveitis (TINU syndrome) associated with granulomatous hepatitis. AB - A 23-year-old male presented with renal failure, cholestatic liver enzyme elevation and uveitis. Percutaneous renal biopsy revealed marked eosinophilic infiltration of the renal interstitium, which made the diagnosis of TINU syndrome (Tubulo-Interstitial Nephritis and Uveitis). Percutaneous liver biopsy showed granulomatous hepatitis, which was not described as a part of TINU syndrome. The diagnostic dilemma and the literature are discussed. PMID- 10363633 TI - Lupus nephritis in juvenile myelomonocytic leukemia. AB - A 13-year-old girl developed lupus nephritis and Hashimoto thyroiditis in the chronic phase of juvenile myelomonocytic leukemia (JMML). At age 7 months, she was diagnosed as having JMML based on the hepatosplenomegaly, leukocytosis, thrombocytopenia, increased levels of fetal hemoglobin, and spontaneous in vitro growth of granulocyte-macrophage progenitors. At the onset of JMML, she had hypergammaglobulinemia, antinuclear antibodies, rheumatoid factors and anti smooth muscle antibody. She had been placed on oral 6-mercaptopurine for about 12 years, with clinical improvement. At age 13 years, she was found to have hematuria and proteinuria. She also developed arthritis and Raynaud's phenomenon as well. She had antinuclear antibodies, rheumatoid factors, LE phenomenon, beta 1C (C3) nephritic factor (C3NeF), antithyroid antibodies, and hypocomplementemia. The renal biopsy specimens revealed a diffuse increase in the mesangial cells and matrix by light microscopy, and intense staining of IgG, Clq and C3 by immunofluorescence microscopy. The hormonal study ultimately showed decreased thyroid functions. So she was diagnosed as lupus nephritis and Hashimoto thyroiditis. The patient is the first example to show close relationship between stem cell abnormalities in JMML and development of overt autoimmune disorders. PMID- 10363634 TI - Goodpasture's syndrome with normal renal function. PMID- 10363635 TI - Hepatocellular carcinoma among hemodialysis patients infected with hepatitis C virus--early evolution and rapid progression. PMID- 10363636 TI - Brown tumor and tumoral calcinosis 6 years after renal transplantation. PMID- 10363637 TI - The possibility of captopril and aspirin test for the diagnosis of patients with renovascular hypertension. PMID- 10363638 TI - An unusual case of dense deposit disease with nodular sclerotic lesions of the glomeruli. PMID- 10363639 TI - The proteasome-dependent proteolytic system. AB - The 20S proteasome is an intriguingly large complex that acts as a proteolytic catalytic machine. Accumulating evidence indicates the existence of multiple factors capable of regulating the proteasome function. They are classified into two different categories, one type of regulator is PA700 or PA28 that is reversibly associated with the 20S proteasome to form enzymatically active proteasomes and the other type including a 300-kDa modulator and PI31 indirectly influences proteasome activity perhaps by promoting or suppressing the assembly of the 20S proteasome with PA700 or PA28. Thus, there have been documented two types of proteasomes composed of a core catalytic proteasome and a pair of symmetrically disposed PA700 or PA28 regulatory particle. Moreover, the recently identified proteasome containing both PA28 and PA700 appears to play a significant role in the ATP-dependent proteolytic pathway in cells, as can the 26S proteasome which is known as a eukaryotic ATP-dependent protease. PMID- 10363640 TI - Phosphorylation of proteasomes in mammalian cells. AB - 20S proteasomes are large (700 kDa) proteinase complexes which form the central catalytic core of larger complexes (26S proteasomes or PA28-20S complexes) formed by association with regulatory particles. These larger complexes are involved in diverse regulatory processes in the cell including cyclin breakdown, proteolytic control of transcription factors and other short-lived regulatory proteins, and antigen presentation. In order to carry out these diverse functions the proteasome complexes must be held under tight regulatory control. The early recognition of potential phosphorylation sites in a number of core and regulatory subunits suggested that some control of the complexes activities may be via phosphorylation. We have investigated the role of phosphorylation in determining proteasome localization, activities and association with regulatory complexes. PMID- 10363641 TI - Assembly of the regulatory complex of the 26S proteasome. AB - The 19S regulatory complex (RC) of 26S proteasomes is a 900-1000 kDa particle composed of 18 distinct subunits (S1-S15) ranging in molecular mass from 25 to 110 kDa. This particle confers ATP-dependence and polyubiquitin (polyUb) recognition to the 26S proteasome. The symmetry and homogenous structure of the proteasome contrasts sharply with the remarkable complexity of the RC. Despite the fact that the primary sequences of all the subunits are now known, insight has been gained into the function of only eight subunits. The six ATPases within the RC constitute a subfamily (S4-like ATPases) within the AAA superfamily and we have shown that they form specific pairs in vitro. We have now determined that putative coiled-coils within the variable N-terminal regions of these proteins are likely to function as recognition elements that direct the proper placement of the ATPases within the RC. We have also begun mapping putative interactions between non-ATPase subunits and S4-like ATPases. These studies have allowed us to build a model for the specific arrangement of 9 subunits within the human regulatory complex. This model agrees with recent findings by Glickman et al. who have reported that two subcomplexes, termed the base and the lid, form the RC of budding yeast 26S proteasomes. PMID- 10363642 TI - Functional analysis of the proteasome regulatory particle. AB - We have developed S. cerevisiae as a model system for mechanistic studies of the 26S proteasome. The subunits of the yeast 19S complex, or regulatory particle (RP), have been defined, and are closely related to those of mammalian proteasomes. The multiubiquitin chain binding subunit (S5a/Mcb1/Rpn10) was found, surprisingly, to be nonessential for the degradation of a variety of ubiquitin protein conjugates in vivo. Biochemical studies of proteasomes from deltarpn10 mutants revealed the existence of two structural subassemblies within the RP, the lid and the base. The lid and the base are both composed of 8 subunits. By electron microscopy, the base and the lid correspond to the proximal and distal masses of the RP, respectively. The base is sufficient to activate the 20S core particle for degradation of peptides, but the lid is required for ubiquitin dependent degradation. The lid subunits share sequence motifs with components of the COP9/signalosome complex, suggesting that these functionally diverse particles have a common evolutionary ancestry. Analysis of equivalent point mutations in the six ATPases of the base indicate that they have well differentiated functions. In particular, mutations in one ATPase gene, RPT2, result in an unexpected defect in peptide hydrolysis by the core particle. One interpretation of this result is that Rpt2 participates in gating of the channel through which substrates enter the core particle. PMID- 10363643 TI - Comparison of human COP9 signalsome and 26S proteasome lid'. AB - The human core COP9 signalosome consists of eight subunits which have been identified, cloned and sequenced. The components of COP9 signalosome possess homologies with eight non-ATPase regulatory subunits of the 26S proteasome. These polypeptides of the 19S regulator form a reversibly binding subcomplex called the 'lid'. We isolated the 'lid' from human red blood cells and compared it with the COP9 signalosome complex. In addition to the non-ATPase regulatory polypeptides, we found a high molecular mass ATPase copurifying with the human 'lid'. The COP9 signalosome-associated kinase activity is either not at all or only weakly affected by common kinase inhibitors such as 1-(5-Isoquinolinesulfonyl)-2-methyl piperazine (H7), 5,6-dichloro-1-beta-D-ribofuranosyl-benzimidazole (DRB) or Wortmannin. Curcumin, a tumor suppressor and effector of AP-1 activation, is a potent inhibitor of the COP9 signalosome kinase activity with a Ki of about 10 microM. Since curcumin is known as an inhibitor of the c-Jun N-terminal kinase (JNK) signaling pathway acting upstream of the MAP kinase kinase kinase level, one site of action of the COP9 signalosome might be proximal to regulators on that level. PMID- 10363644 TI - Activator complexes containing the proteasomal regulatory ATPases S10b (SUG2) and S6 (TBP1) in different tissues and organisms. AB - Each 19S regulator of the 26S proteasome contains six ATPase subunits as well as many (>14) non-ATPase protein subunits. The ATPase subunits have been detected in other complexes which may regulate transcription and possibly other cellular processes. The S10b (yeast SUG2 or human p42) and the S6' (TBP1) ATPases have been found in an activator complex (modulator) prepared from bovine red cells. We have identified and partially characterised a similar activator from different human tissues (from soluble extracts of human brain, placenta and human embryonic kidney cells) and an insect: an activator is present in soluble extracts of abdominal intersegmental muscle from Manduca sexta. Activation is ATP and concentration dependent. There is no stimulation of human red cell-derived 20S proteasome by the Manduca activator ruling out 11S regulator in the preparations. Additionally, cross-species activation occurs: the Manduca activator increases the activity of rat skeletal muscle 26S proteasomes and the human placental activator similarly increases the activity of 26S proteasomes prepared from muscles from Manduca sexta. Finally, there is no evidence for other ATPases in the activator complex. PMID- 10363645 TI - Modulators of the activation of the proteasome by PA28 (11S reg). AB - The degradation of chromogenic substrates and oligopeptides by the 20S proteasome is markedly enhanced and the generation of antigens for presentation by the MHC class-I system is facilitated by combination with an activator protein known as PA28 or 11S reg. We have described the properties of a PA28-proteasome modulator, N-benzyloxycarbonyl-Ile-Glu(O-t-Bu)-Ala-leucinol which shifts the pathway of peptide hydrolysis by the activated proteasome to products terminating in an acidic amino acid at the expense of products terminating in a hydrophobic amino acid. We now report that piperazinyl phenothiazines and several other antipsychotic drugs modulate the PA28-20S activated proteasome in an opposite manner. Fluphenazine, trifluoperazine and prochlorperazine antagonize the peptidylglutamyl peptide bond hydrolyzing activity of the activated proteasome much more strongly than the chymotrypsinlike activity. The chicken ovalbumin immunodominant epitope SIINFEKL is degraded by the activated proteasome to SIINFE and SIINF in approximately equimolar amounts. Piperazinyl phenothiazines promote formation of SIINF whereas Psi-ol promotes formation of SIINFE. PA28- proteasome modulators by modifying the profile of peptides produced by the activated proteasome, may either enhance or suppress the immune response. PMID- 10363646 TI - Are there multiple proteolytic pathways contributing to c-Fos, c-Jun and p53 protein degradation in vivo? AB - The c-Fos and c-Jun oncoproteins and the p53 tumor suppressor protein are short lived transcription factors. Several catabolic pathways contribute to their degradation in vivo. c-Fos and c-Jun are thus mostly degraded by the proteasome, but there is indirect evidence that, under certain experimental/physiological conditions, calpains participate in their destruction, at least to a limited extent. Lysosomes have also been reported to participate in the destruction of c Fos. Along the same lines, p53 is mostly degraded following the ubiquitin/proteasome pathway and calpains also seem to participate in its degradation. Moreover, c-Fos, c-Jun and p53 turnovers are regulated upon activation of intracellular signalling cascades. All taken together, these observations underline the complexity of the mechanisms responsible for the selective destruction of proteins within cells. PMID- 10363647 TI - Proteasome-dependent degradation of human CDC25B phosphatase. AB - The CDC25 dual specificity phosphatase is a universal cell cycle regulator. The evolutionary conservation of this enzyme from yeast to man bears witness to its major role in the control of cyclin-dependent kinases (CDK) activity that are central regulators of the cell cycle machinery. CDC25 phosphatase both dephosphorylates and activates CDKs. Three human CDC25s have been identified. CDC25A is involved in the control of G1/S, and CDC25C at G2/M throught the activation of CDK 1-cyclin B. The exact function of CDC25B however remains elusive. We have found that CDC25B is degraded by the proteasome pathway in vitro and in vivo. This degradation is dependent upon phosphorylation by the CDK1 cyclin A complex, but not by CDK1-cyclin B. Together with the observations of others made in yeast and mammals, our results suggest that CDC25B might act as a 'mitotic starter' triggering the activation of an auto-amplification loop before being degraded. PMID- 10363648 TI - Degradation of MyoD by the ubiquitin pathway: regulation by specific DNA-binding and identification of a novel site for ubiquitination. AB - MyoD is a tissue-specific transcriptional activator involvd in skeletal muscle differentiation. It is induced during transition from proliferating, non differentiated myoblasts to the resting and well differentiated myotubes. Like many other transcriptional regulators, it is short-lived, however, the targeting proteolytic pathway and the underlying regulatory mechanisms involved have remained obscure. Here we show that MyoD is degraded by the ubiquitin system both in vivo and in vitro. In cells, degradation is inhibited by lactacystin, a specific inhibitor of the 20S proteasome. Inhibition is accompanied by accumulation of MyoD-ubiquitin conjugates. In a cell free system, the proteolytic process requires both ATP and ubiquitin and is preceded by formation of MyoD ubiquitin adducts. Interestingly, the process is inhibited by the specific DNA sequence to which MyoD binds. Analysis of the ubiquitination site has revealed that the N-terminal residue of MyoD is sufficient and essential to promote conjugation and subsequent degradation of the protein: conjugation to internal Lys residues is not necessary. Substitution of all Lys residues did not affect significantly its degradation either in intact cells or in a reconstituted cell free system. Degradation was inhibited by specific proteasome inhibitors and was accompanied by accumulation of ubiquitinated species of the protein. We concluded that the first ubiquitin moiety is attached via its C-terminal Gly to the N terminal residue of MyoD, and the polyubiquitin chain is then synthesized on Lys48 of this moiety. PMID- 10363649 TI - The ubiquitin/proteasome pathway: friend or foe in zinc-, cadmium-, and H2O2 induced neuronal oxidative stress. AB - One of the hallmarks of neurodegeneration is the accumulation of ubiquitinated proteins in intraneuronal inclusions in the cytosol, endosomes/lysosomes and nuclei of affected cells. The relationship between inclusion production and cell viability is not well understood. On the one hand inclusions may be beneficial and result from an attempt of the cell to isolate a subclass of ubiquitinated proteins that are not effectively degraded. On the other hand, the inclusions may impede normal cell function contributing to cell death. To address this issue we treated mouse neuronal HT4 cells with three toxic agents cadmium, zinc and H2O2, and investigated their effects on glutathione homeostasis, on accumulation of ubiquitinated proteins and on cell viability. The three treatments induce oxidative stress manifested by decreases in glutathione (GSH) and/or increases in protein mixed disulfides (PrSSG). After an overnight recovery period in the absence of treatment, GSH and PrSSG were restored to almost normal levels. However, the levels of ubiquitinated proteins were significantly increased, and cell viability was sharply reduced. These results suggest that the ubiquitin proteasome pathway is recruited for removal of proteins that are oxidatively modified. However, if the ubiquitinated proteins are not efficiently degraded, they accumulate in the cell and contribute to a decrease in cell viability. PMID- 10363651 TI - Adaptation of the ubiquitin-proteasome proteolytic pathway in cancer cachexia. AB - The ubiquitin-proteasome proteolytic pathway is of major importance in the breakdown of skeletal muscle proteins. The first step in this pathway is the covalent attachment of polyubiquitin chains to the targeted protein. Polyubiquitinylated proteins are then recognized and degraded by the 26S proteasome complex. In this review, we critically analyze recent findings in the regulation of ubiquitinylation of protein substrates and of their subsequent proteasome-dependent degradation in animal models of cancer cachexia. In particular, we discuss the influence of various mediators (anorexia, hormones, prostaglandins, cytokines, and proteolysis-inducing factor) in signaling the activation of ubiquitin-proteasome proteolysis in skeletal muscle. These findings have lead to new concepts that are starting to be used for preventing cachexia in cancer and other wasting diseases. PMID- 10363650 TI - Role of the ubiquitin-proteasome pathway in sepsis-induced muscle catabolism. AB - Several lines of evidence suggest that the ubiquitin-proteasome pathway is involved in sepsis-induced muscle catabolism. The gene expression of ubiquitin and several of the proteasome subunits was increased in muscle from both septic rats and patients. In other studies, the activity of isolated 20S proteasomes was stimulated in septic muscles. Sepsis-induced increase in muscle total and myofibrillar protein breakdown was inhibited with specific proteasome blockers. Although the ubiquitin-proteasome pathway is upregulated in septic muscle, it is still unclear how the myofibrillar proteins actin and myosin are ubiquitinated and become substrates for the 26S proteasome. Recent studies suggest that a calcium-dependent, calpain-mediated process releases myofilaments from the Z disks during sepsis. It is possible that this process exposes destabilizing N terminal residues on actin and myosin, making them suitable substrates for the N end rule pathway involving the 14 kD ubiquitin-conjugating enzyme E214k and the ubiquitin-protein ligase E3alpha. PMID- 10363652 TI - Alterations of proteasome activities in skeletal muscle tissue of diabetic rats. AB - During the last years many investigations have shown that a major catalyst within the mechanism of skeletal muscle wasting occurring under conditions like sepsis, injuries, trauma, cancer cachexia, chronic acidosis, fasting, glucocorticoid treatment, and insulinopenia is the ubiquitin-proteasome system. Evidence for this was obtained by findings that the rate of ATP-dependent protein degradation is increased, that m-RNA concentrations of several proteasome subunits and ubiquitin are increased and the amount of ubiquitin-protein conjugates is elevated under these conditions. Additionally, the enhanced protein breakdown was shown to be suppressed by proteasome inhibitors. In the present report we show that most but not all of the proteolytic activities of partially purified 20S/26S proteasomes from skeletal muscle of rats increase after induction of Diabetes mellitus. This finding suggests that part of the mechanism of acceleration of muscle protein breakdown is due to changes in proteasome activities. PMID- 10363653 TI - Changes in 20S proteasome activity during ageing of the LOU rat. AB - Muscular functions decline and muscle mass decreases during ageing. In the rat, there is a 27% decrease in muscle protein between 18 and 34 months of age. We examined age-related changes in the proteasome-dependent proteolytic pathway in rats at 4, 18, 24, 29 and 34 months of age. The three best characterised activities of the proteasome (chymotrypsin-like, trypsin-like and peptidylglutamyl peptide hydrolase) increased to 29 months and then decreased in the senescent animal. These variations in activity were accompanied by an identical change in the quantity of 20S proteasome measured by Western blot, whereas the S4 subunit of the 19S regulator and the quantity of ubiquitin-linked proteins remained constant. mRNA of subunits C3, C5, C9, and S4 increased in the senescent animal, but ubiquitin mRNA levels were unchanged. These findings suggest that the 20S proteasome may be partly responsible for the muscular atrophy observed during ageing in the rat. PMID- 10363654 TI - Manipulation of the ubiquitin-proteasome pathway in cachexia: pentoxifylline suppresses the activation of 20S and 26S proteasomes in muscles from tumor bearing rats. AB - The development of pharmacological approaches for preventing the loss of muscle proteins would be extremely valuable for cachectic patients. For example, severe wasting in cancer patients correlates with a reduced efficacy of chemotherapy and radiotherapy. Pentoxifylline (PTX) is a very inexpensive xanthine derivative, which is widely used in humans as a haemorheological agent, and inhibits tumor necrosis factor transcription. We have shown here that a daily administration of PTX prevents muscle atrophy and suppresses increased protein breakdown in Yoshida sarcoma-bearing rats by inhibiting the activation of a nonlysosomal, Ca(2+) independent proteolytic pathway. PTX blocked the ubiquitin pathway, apparently by suppressing the enhanced expression of ubiquitin, the 14-kDa ubiquitin conjugating enzyme E2, and the C2 20S proteasome subunit in muscle from cancer rats. The 19S complex and 11S regulator associate with the 20S proteasome and regulate its peptidase activities. The mRNA levels for the ATPase subunit MSS1 of the 19S complex increased in cancer cachexia, in contrast with mRNAs of other regulatory subunits. This adaptation was suppressed by PTX, suggesting that the drug inhibited the activation of the 26S proteasome. This is the first demonstration of a pharmacological manipulation of the ubiquitin-proteasome pathway in cachexia with a drug which is well tolerated in humans. Overall, the data suggest that PTX can prevent muscle wasting in situations where tumor necrosis factor production rises, including cancer, sepsis, AIDS and trauma. PMID- 10363655 TI - Structure and functions of arthropod proteasomes. AB - Recent work on structural/functional relationships in arthropod proteasomes is reviewed. Taking advantage of our ability to induce a stable, proteolytically active conformation of the lobster proteasome, the structures of basal and heat activated complexes were probed with exogenous proteases. Increased sensitivity to chymotrypsin and trypsin showed that heat activation induced a more 'open' conformation, allowing entry of large substrates into the catalytic chamber. In Drosophila, the effects of two developmental mutant alleles (DTS-7 and DTS-5) encoding proteasome subunits (Z and C5, respectively) on the subunit composition and catalytic activities of the enzyme were examined. Both qualitative and quantitative differences in compositions between wild-type (+/+) and heterozygotes (+/DTS) indicated that incorporation of mutant subunits alters post translational modifications of the complex. Catalytic activities, however, were similar, which suggests that the developmental defect involves other proteasome properties, such as intracellular localization and/or interactions with endogenous regulators. A hypothetical model in which DTS subunits act as poison subunits is presented. PMID- 10363656 TI - Relationships between proteasomes and viral gene products. AB - The interrelationships between proteasomes and viral gene products are very complex. 20S proteasomes associate with a number of viral mRNAs which are cleaved by proteasome's associated endonuclease activity. In addition proteasome's endopeptidase activities are involved in the presentation of viral antigens. Viral proteins of different origin associate with the 20S and 26S complexes and interfere with their enzymatic activities. A major part of this review deals with the interactions between 20S proteasomes and the gene products of the human immunodeficiency virus (HIV) which has been studied in detail by our group. PMID- 10363657 TI - Proteasome subunit zeta, a putative ribonuclease, is also found as a free monomer. AB - 20 S Proteasomes are large proteinase complexes found in eukaryotic cells where they degrade cell proteins in an ATP-dependent manner. Proteasomes consist of 14 different subunits. One of them, zeta, was found in HeLa cells at a concentration of 890 microg per g of cell protein. A large proportion of zeta was found in the free state rather than incorporated into proteasomes, namely 28% in HeLa cells and 37% in BSC-1 cells. Free zeta was found in both nuclei and cytoplasm. In HeLa cells free zeta had a t1/2 of 2.8 h, compared to 5 d for proteasomes, and did not exchange with zeta in proteasomes. We confirmed (Petit F et al.: Biochem. J. 326: 93-98 (1997)) that both 20 S proteasomes and free zeta subunits possess RNase activity though the activities were very low: 4 mMoles and 0.6 mMoles of tobacco mosaic virus RNA degraded per mole of enzyme per min, respectively. The physiological function of the relatively abundant zeta monomers is not known. PMID- 10363658 TI - Endoplasmic reticulum degradation. Reverse protein transport and its end in the proteasome. AB - Degradation of misfolded or unassembled proteins of the secretory pathway is an essential function of the quality control system of the Endoplasmic Reticulum (ER). Using yeast as a model organism we show that a mutated and therefore misfolded soluble lumenal protein carboxypeptidase yscY (CPY), and a polytopic membrane protein, the ATP-binding cassette transporter Pdr5 (Pdr5), are retrograde transported out of the ER and degraded via the cytoplasmic ubiquitin proteasome system. Retrograde transport depends on an intact Sec61 translocon. Complete import of CPY into the lumen of the ER requests a new targeting mechanism for retrograde transport of the malfolded enzyme through the Sec61 channel to occur. For soluble CPY, but not for the polytopic membrane protein Pdr5 action of the ER-lumenal Hsp70 chaperone Kar2 is necessary to deliver the protein to the ubiquitin-proteasome machinery. Polyubiquitination of CPY and Pdr5 by the ubiquitin conjugating enzymes Ubc6 and Ubc7 is crucial for degradation to occur. Also transport of CPY out of the ER-lumen depends on ubiquitination. Newly discovered proteins of the ER membrane, Derl, Der3/Hrd1, and Hrd3 are specifically involved in the retrograde transport processes. PMID- 10363660 TI - Structure and functional analysis of the 26S proteasome subunits from plants. AB - As initial steps to define how the 26S proteasome degrades ubiquitinated proteins in plants, we have characterized many of the subunits that comprise the proteolytic complex from Arabidopsis thaliana. A set of 23 Arabidopsis genes encoding the full complement of core particle (CP) subunits and a collection encoding 12 out of 18 known eukaryotic regulatory particle (RP) subunits, including six AAA-ATPase subunits, were identified. Several of these 26S proteasome genes could complement yeast strains missing the corresponding orthologs. Using this ability of plant subunits to functionally replace yeast counterparts, a parallel structure/function analysis was performed with the RP subunit RPN 10/MCB1, a putative receptor for ubiquitin conjugates. RPN10 is not essential for yeast viability but is required for amino acid analog tolerance and degradation of proteins via the ubiquitin-fusion degradation pathway, a subpathway within the ubiquitin system. Surprisingly, we found that the C terminal motif required for conjugate recognition by RPN10 is not essential for in vivo functions. Instead, a domain near the N-terminus is required. We have begun to exploit the moss Physcomitrella patens as a model to characterize the plant 26S proteasome using reverse genetics. By homologous recombination, we have successfully disrupted the RPN10 gene. Unlike yeast rpn10delta strains which grow normally, Physcomitrella rpn10delta strains are developmentally arrested, being unable to initiate gametophorogenesis. Further analysis of these mutants revealed that RPN10 is likely required for a developmental program triggered by plant hormones. PMID- 10363659 TI - GFP-labelling of 26S proteasomes in living yeast: insight into proteasomal functions at the nuclear envelope/rough ER. AB - 26S proteasomes are multisubunit protease complexes that play the central role in the ubiquitin-dependent protein degradation pathway. The proteolytically active core is formed by the 20S proteasome. Regulatory subunits, principally the 19S cap complex, confer the specificity towards ubiquitinated substrates and an ATP dependence on proteolysis. Green fluorescence protein (GFP)-tagged versions of either an alpha-subunit of the 20S core or an ATPase subunit of the 19S cap complex were functionally incorporated into the protease complex, thus allowing to monitor the subcellular distribution of 26S proteasomes in living yeast. Our localization studies suggest that proteasomal proteolysis mainly occurs at the nuclear envelope (NE)/rough ER. Implications of proteasomal functions at the NE/rough ER are discussed in the context of published work on ER degradation and with regard to possible targeting mechanisms. PMID- 10363661 TI - Sub-grouping non-melancholic depression from manifest clinical features. AB - AIM: To determine whether clinical symptoms manifested during an episode of major depression are sufficient to allow meaningful sub-groups of non-melancholic depression to be identified. METHODS: A sample of 178 non-melancholic patients with a major depressive episode was studied. The initial set of clinical variables was refined to 38 (21 depression, 17 anxiety) items and a cluster analysis undertaken. RESULTS: A four-cluster solution identified 'anxiety', 'irritability', 'depressed mood' and 'residual' clusters, with these labels clarified by reference to a large data bank of non-symptomatic variables. These analyses suggested that members of the first two clusters could be viewed as having spectrum conditions (whereby Axis I symptom states are able to be linked with precursor or prodromal states and personality). CONCLUSIONS: We confirm the long-standing suggestion that the non-melancholic depressive class contains sub groups of those with manifest states of anxious depression and of an irritable or 'hostile' depression, and that such manifest symptoms are likely to be rooted in and fed by temperament and personality characteristics. The delineation of such sub-groups should assist studies designed to identify underlying neurobiological underpinnings and clinical management of the non-melancholic depressive disorders. PMID- 10363662 TI - Selective serotonin reuptake inhibitors have broadened the utilisation of antidepressant treatment in accordance with recommendations. Findings from a Swedish prescription database. AB - BACKGROUND: With the advent of the selective serotonin reuptake inhibitors (SSRIs), the use of antidepressants has increased drastically in Sweden. The use of tricyclic antidepressants (TCAs) has, however, decreased. METHODS: We surveyed a prescription database in the Swedish county of Jamtland and compared prescription patterns for patients prescribed SSRIs with those prescribed TCAs. RESULTS: The incidence of treatments of antidepressants increased from 0.76% to 1.33% during the period 1991-1996. There were no significant differences between SSRIs and TCAs with regard to patients having only one prescription dispensed within three months from the index prescription, or patients who switched class of antidepressant. Only a minority of the treatments were continued for at least six months, but significantly more SSRI than TCA treatments (42% and 27%). A second treatment period suggesting recurrence was three-times more common in the TCA group than in the SSRI group. CONCLUSION: Provided that the increased use of SSRIs is mainly for depression, these drugs appear, despite a lower efficacy in severe depression, to have enabled a broader utilisation of antidepressants with regard to incidence, dosage and duration, in accordance with recommendations. Further analyses of this phenomenon relative to diagnostic criteria and outcome measures are required. PMID- 10363663 TI - Accelerated post-glucose glycaemia and altered alliesthesia-test in Seasonal Affective Disorder. AB - BACKGROUND: Little is known about the link between mood, food and metabolic function in Seasonal Affective Disorder (SAD). METHODS: We investigated this link in a combined glucose tolerance-alliesthesia test in eight SAD patients in winter before and after one week light therapy, and in summer. RESULTS: SAD patients exhibited faster post-glucose glycaemic and insulin responses (p <0.05), and increased hedonic ratings of high concentrated sucrose solutions (p <0.035) when depressed in winter than when euthymic (one week after light treatment or in summer). CONCLUSIONS: The rapid glycaemic and insulin responses to an oral glucose load may be a result of accelerated gastric emptying. LIMITATIONS: The number of studied patients was rather small and no control group was studied in parallel. CLINICAL RELEVANCE: the more rapid post-glucose glycaemia may impair glucose homeostasis in depressed SAD patients. PMID- 10363664 TI - Higher serum prolyl endopeptidase activity in patients with post-traumatic stress disorder. AB - BACKGROUND: It is reported that psychiatric disorders, such as depression and schizophrenia, are associated with changes in serum activity of prolyl endopeptidase (EC 3.4.21.26), a cytosolic endopeptidase, which cleaves peptide bonds on the carboxylside of proline in proteins of relatively small molecular mass. AIMS AND METHODS: The aims of the present study were to examine serum PEP activity in patients with post-traumatic stress disorder (PTSD) versus healthy volunteers. PEP activity has been determined by a fluorimetric assay. RESULTS: Serum PEP activity was significantly higher in patients with PTSD than in normal volunteers. Serum PEP activity was significantly higher in patients with PTSD and concurrent major depression than in patients with PTSD without major depression. In PTSD patients, there were no significant correlations between serum PEP activity and severity of PTSD symptoms. CONCLUSIONS: The results show that PTSD and, in particular, PTSD with concurrent major depression is associated with increased activity of PEP. RELEVANCE: these results may be of importance for the (i) neuroendocrine pathophysiology of PTSD since PEP degrades neuropeptides, such as arginine vasopressin (AVP) and thyrotropin releasing hormone (TRH); and (ii) etiology of PTSD, since PEP degrades behaviorally active neuropeptides, such as AVP, TRH, oxytocin, neurotensin and substance P, which play a key role in positive reinforcement, social interactions, emotions and stress responsivity. PMID- 10363665 TI - Seasonal affective disorder and latitude: a review of the literature. AB - BACKGROUND: The aim of the study is to investigate the relationship between the prevalence of SAD and latitude. METHODS: An overview of the epidemiological literature on the prevalence of SAD is given and studies relevant for the latitudinal dependency of prevalence will be analyzed and discussed. RESULTS: The mean prevalence of SAD is two times higher in North America compared to Europe. Over all prevalence studies, the correlation between prevalence and latitude was not significant. A significant positive correlation was found between prevalence and latitude in North America. For Europe there was a trend in the same direction. CONCLUSIONS: The influence of latitude on prevalence seems to be small and other factors like climate, genetic vulnerability and social-cultural context can be expected to play a more important role. Additional controlled studies taking these factors into account are necessary to identify their influence. PMID- 10363666 TI - The effect of comorbid alcoholism on recurrence in affective disorder: a case register study. AB - BACKGROUND: Studies of the effect of comorbid alcoholism on the risk of recurrence in affective disorder have given contradictory results. METHOD: Using survival analysis, the rate of recurrence was calculated in a case register study including all hospital admissions with primary affective disorder in Denmark during 1971-1993. The rate of recurrence was estimated following each new affective episode. RESULTS: In all, 20 350 patients were discharged after first admission with a main diagnosis of affective disorder of depressive or manic/circular type. Among these, 518 patients (2.6%) had an auxiliary diagnosis of alcoholism. Patients with a current auxiliary diagnosis of alcoholism had increased rate of recurrence following the first three affective episodes but not following subsequent episodes compared with patients without auxiliary diagnoses. The effect of alcoholism declined with the number of episodes. In contrast, no effect was found of other auxiliary diagnoses on the rate of recurrence. CONCLUSION: Rehospitalisation data suggest that concurrent alcoholism increases the risk of recurrence of affective episodes during the initial course of unipolar and bipolar disorder but has no effect on recurrence later in the course of the illnesses. LIMITATION: The data relate to re-admissions rather than recurrence. CLINICAL RELEVANCE: The study emphasises that alcoholism has a deteriorating effect on the course of affective disorder. PMID- 10363667 TI - The development and validation of the coping inventory for prodromes of mania. AB - This study described the construction and validation of the Coping Inventory for Prodromes of Mania (CIPM)-a newly developed instrument aimed at assessing how manic depressive sufferers dealt with their prodromes of mania. Two hundred and six subjects completed a 40-item measure of coping responses to prodromes of mania, a measure of social functioning and a mood measure. The CIPM yielded four empirically derived subscales: stimulation reduction, problem-directed coping, seeking professional help, denial or blame. All of these subscales provided good reliability and acceptably high internal consistency data. There was also some evidence of good validity of the CIPM. PMID- 10363668 TI - Reliability of biographical data, their relations to personality variables and their influence on life-events. AB - BACKGROUND: First, this study aims to investigate the reliability of biographical and personality data, i.e. analysing the impact of depressive mood on these variables. Second, the influence of early life experience and personality on the reporting of life events was examined. METHODS: Self-reporting questionnaires were administered for a sample of 250 depressive subjects at the beginning, and to assess the influence of depressive mood on the reporting of biographic data at the end of inpatient treatment we used a random sample of 50 subjects out of the 250 patients. RESULTS: Biographical data, unlike personality data, are not significantly influenced by depressive mood and depressive cognition. The number of life-events, and their mean subjective stress, neuroticisms and aim relatedness are on a direct path strongly influenced by biographical data. That means that the more negatively primary socialisation was reported, the more life events were communicated, with corresponding increases in the reporting of their subjective stress, and more neuroticism and less aim-relatedness, and vice versa. Neuroticism strongly influences in a positive way, and aim-relatedness negatively influences in a medium way, the number of life-events and their subjective stress. That means the higher neuroticism and the lower aim-relatedness were reported, the more life-events and higher stress were communicated, and vice versa. CONCLUSION: Linear causal processes from mood-independent factors (e.g. biographical factors, vulnerability) may be the beginning of cyclic causal processes, i.e. vicious circles between life-events and mood-dependent factors (e.g. personality variables). LIMITATIONS: According to the design of the investigation there is no differentiation possible between personality and depression. CLINICAL RELEVANCE: To avoid vicious circles between life-events and mood-dependent factors, preventive psychotherapeutic intervention seems to be necessary to avoid the genesis of harmful life-events. PMID- 10363669 TI - Seasonal and circadian rhythms in suicide in Cagliari, Italy. AB - BACKGROUND: In previous studies of the seasonality of suicide, peaks have often been found in the number of suicides in the spring and early summer in both northern and southern hemispheres. The purpose of the present study was to investigate the distribution of suicide as to month, seasons, day of the week, and time of the day. METHOD: Data on suicides in Cagliari (Italy) in the period 1990-1994 were analyzed by means of spectral analysis, cosinor and multiple regression analysis. RESULTS: Two seasonal rhythms, i.e. an annual and a semiannual rhythm, accounted for 25% of the variation in the total number of suicides. The peak number of suicides occurred in February with a second less significant peak in June and July. Lows were found in November and December. There were no significant differences in number of suicides in relation to days of the week. Three rhythms, i.e. 24 hours (circadian), 8 hours and 1 hour, explained 63.9% of the variance in the number of suicides by time of the day. Peak numbers in number of suicides were found between 08:31 and 12:30h, while the number of suicides was also significantly higher between 12:31h and 20:30h than between 20.31h and 8.30h. Age and gender did not significantly affect the seasonal and circadian rhythms in suicide. CONCLUSIONS: The results show that there is a significantly seasonal variation and a highly significant variation by time of the day in suicide in Cagliari, Italy. PMID- 10363670 TI - Low persistence in euthymic manic-depressive patients: a replication. AB - BACKGROUND: A previous study which compared euthymic Israeli bipolars in a public hospital clinic to US normative data suggested that low Persistence scores on the Tridimensional Personality Questionnaire (TPQ) may be a temperament marker for manic-depressive illness. The current study attempts to replicate that finding. METHODS: A new sample of 25 euthymic Israeli bipolars in private treatment was compared to Israeli normal controls matched for gender and ethnic background. All subjects completed Hebrew versions of the TPQ. RESULTS: Patients scored lower than controls on the Novelty Seeking and Persistence Scales. LIMITATIONS AND CONCLUSIONS: Though sample and effect size are both small, the finding of low Persistence is consistent with that of the earlier study. PMID- 10363671 TI - Characteristics of patients with otherwise typical winter depression, but with incomplete summer remission. AB - BACKGROUND: Seasonal affective disorder, winter depression type (WD-SAD), is characterized by recurring autumn/winter depression with full remission or hypomania/mania in summer. However, some patients have an otherwise typical WD but with incomplete summer remission. We wanted to elucidate in what other respects such patients differ from typical WD-SAD patients. METHODS: 14 patients meeting DSM-III-R criteria for Seasonal Pattern except for incomplete summer remission (ISR), were compared with 144 patients meeting the full criteria, including complete summer remission (CSR), with regard to demography, illness history, clinical symptoms, and response to light treatment. RESULTS: In comparison with the CSR group, the ISR group had a longer duration of illness, more often used antidepressants, and improved significantly less after treatment with bright light for 6 days, whereas the symptomatology in winter (Montgomery Asberg Depression Rating Scale plus hypersomnia, hyperphagia, and carbohydrate craving) was similar in the two groups. LIMITATIONS: The ISR group was small, and the severity of their summer depression could only be assessed retrospectively. CONCLUSIONS: Patients with otherwise typical WD but with incomplete summer remission respond poorly to light treatment. Full summer remission should be retained as a criterion for WD-SAD. PMID- 10363672 TI - Lamotrigine for the treatment of bipolar disorder: a clinical case series. AB - BACKGROUND: Recently, a number of new agents have become available to treat bipolar disorder, however many patients may not respond fully even when used in combination. Early reports in epilepsy studies suggested mood-related effects of lamotrigine treatment, as have preliminary reports in bipolar patients. METHODS: Seventeen patients meeting DSM-IV criteria for bipolar I (n = 9) or bipolar II (n = 8) disorder displaying affective symptoms and a past history of inadequate response or tolerability to at least two standard mood stabilizing agents were recruited through the Stanley Foundation Bipolar Network and treated with the new anticonvulsant lamotrigine in an add-on, open-label study. Response to therapy was assessed using the Clinical Global Impression Scale modified for bipolar disorder. RESULTS: The mean dose of lamotrigine was 187+/-157 mg/day (range 50 600 mg/day) for a mean duration of 159+/-109 days (range 14-455 days). Eleven (65%) patients were rated as very much or much improved. Lamotrigine was well tolerated, and may have mood stabilizing and antidepressant properties in some patients with bipolar disorder. LIMITATIONS: The study is hypothesis generating because it was uncontrolled and open. Controlled studies are warranted. CONCLUSIONS: This preliminary report supports clinical improvement for both mood cycling and depression in patients with bipolar disorder treated with lamotrigine. PMID- 10363673 TI - The moon and madness reconsidered. AB - Belief that the full moon is associated with psychiatric disturbance persists despite 50 years research showing no association. This article traces the historical roots of belief in the power of the moon to cause disorders the mind, especially insanity and epilepsy. Putative mechanisms of lunar action are critiqued. It is proposed that modern findings showing lack of lunar effect can be reconciled with pre-modern beliefs in the moon's power through a mechanism of sleep deprivation. Prior to the advent of modern lighting the moon was a significant source of nocturnal illumination that affected sleep-wake cycle, tending to cause sleep deprivation around the time of full moon. This partial sleep deprivation would have been sufficient to induce mania/hypomania in susceptible bipolar patients and seizures in patients with seizure disorders. The advent of modern lighting attenuated this lunar effect, especially in modern urban areas, where most 20th century studies of lunar effects on the mind have been conducted. The hypothesis presented in this article is open to empirical validation or falsification. Potential tests for the sleep-deprivation hypothesis of lunar action are discussed. PMID- 10363674 TI - Molecular nature of a novel bacterial toxin: the cytolysin of Enterococcus faecalis. PMID- 10363675 TI - Induction of antibody responses in the common mucosal immune system by respiratory syncytical virus immunostimulating complexes. AB - Immunostimulating complexes (ISCOMs) containing envelope proteins of respiratory syncytial virus (RSV) were explored as a mucosal delivery system for the capacity of inducing a common mucosal antibody response. Two intranasal (i.n.) administrations of BALB/c mice with ISCOMs induced potent serum IgG, and strong IgA responses to RSV locally in the lungs and the upper respiratory, and remotely in the genital and the intestinal tracts. Virtually no measurable IgA response was found in these mucosal organs after two subcutaneous (s.c.) immunizations. Virus neutralizing (VN) antibodies were detected in serum and in all of the mucosal organ extracts after both s.c. and i.n. immunizations indicating that the neutralizing epitopes were preserved after both mucosal and parenteral modes of administration. While the mucosal IgA response appears to be of mucosal origin, the IgG antibodies to RSV detected in the mucosal organs were likely of serum origin. However, the mucosal VN antibodies correlated with the IgG rather than the IgA levels. An enhanced IgA response to gp120 in various mucosal organs was recorded after i.n. immunization with gp120 incorporated in RSV ISCOMs, indicating a role of RSV envelope proteins in enhancing and targeting mucosal responses to passenger antigens. PMID- 10363676 TI - Inhibition of poliovirus type 1 infection by iron-, manganese- and zinc-saturated lactoferrin. AB - In this study we investigated the inhibitory activity of different milk proteins on poliovirus infection in Vero cells. Proteins analyzed were mucin, alpha lactalbumin, beta-lactoglobulin, and bovine and human lactoferrin. Viral cytopathic effect was not prevented by mucin, alpha-lactalbumin or beta lactoglobulin, whereas the lactoferrins tested were able to inhibit the replication of poliovirus in a dose-dependent manner. Further experiments were carried out in which apo- and native lactoferrin or lactoferrin fully saturated with ferric, manganese or zinc ions were added to the cells during different phases of viral infection. Results obtained demonstrated that all lactoferrins were able to prevent viral replication when present during the entire cycle of poliovirus infection or during the viral adsorption step. Only zinc lactoferrin strongly inhibited viral infection when incubated with the cells after the viral attachment, being the inhibition directly correlated with the degree of zinc saturation. Our results demonstrated that all lactoferrins interfered with an early phase of poliovirus infection; zinc lactoferrin was the sole compound capable of inhibiting a phase of infection subsequent to virus internalization into the host cells. PMID- 10363677 TI - Expression of interferon regulatory factors and indoleamine 2,3-dioxygenase in Chlamydia trachomatis-infected synovial fibroblasts. AB - Synovial fibroblasts probably represent host cells for Chlamydia trachomatis during initial intra-articular infection in reactive arthritis. In vitro synovial cells produce interferon-beta (IFN-beta) in response to chlamydial infection. IFN beta expression can be activated by interferon regulatory factor-1 (IRF-1) and interferon-stimulated gene factor 3gamma (ISGF3gamma). In this study, we demonstrate that infection of synovial fibroblasts with C. trachomatis serotype D induced the expression of IRF-1 mRNA as shown by reverse transcription-PCR. Tumor necrosis factor-alpha (TNF-alpha) stimulation enhanced IRF-1 mRNA levels in infected cells and was required to detect IRF-1 protein by immunoblotting. The level of constitutively expressed IRF-2 was not significantly affected after infection. C. trachomatis was found to cause an up-regulation of ISGF3gamma protein in synovial cells. Induction of the tryptophan-catabolizing enzyme indoleamine 2,3-dioxygenase (IDO) is an important mechanism of the host cell response to control intracellular infection by chlamydiae. It has been described that IRF-1 can induce IDO gene expression. Infection of synovial fibroblasts alone in the absence of exogenous cytokine induced the expression of IDO mRNA which was enhanced by TNF-alpha treatment. The stimulation of IRF-1, ISGF3gamma, and IDO expression was most effective when viable chlamydiae were used as inoculum. Neutralization of IFN-beta in the culture medium of infected cells diminished but did not abrogate expression of IRF-1, ISGF3gamma, and IDO. The increased production of IRF-1 and ISGF3gamma in C. trachomatis-infected synovial fibroblasts may contribute to induction of IFN-beta and IDO. PMID- 10363679 TI - Invasion by Toxoplasma gondii protects human-derived HL-60 cells from actinomycin D-induced apoptosis. AB - Intracellular microorganisms have to rely on the integrity of their host cells to persist. We, therefore, investigated the effect of infections with different Toxoplasma gondii strains on apoptosis of human-derived HL-60 cells at the single cell level. Infection with either mouse-avirulent (NTE strain) or virulent parasites (RH strain) did not induce apoptosis of HL-60 cells as compared to uninfected controls. In contrast, treatment with actinomycin D (act D) led to apoptosis in 15-25% of the cells. However, concomitant infection with T. gondii clearly abrogated act D-induced apoptosis. This was especially apparent in those host cells that were actually infected; in these parasite-positive cells the rate of apoptosis decreased by 82.8+/-4.3% (mean+/-SEM, P=0.017, Student's t-test) and 91.7+/-3.4% (P= 0.024) after infection with either the NTE or the RH strain, respectively. Inhibition of host cell apoptosis was similarly observed in cells which had been invaded by UV-irradiated, non-replicating parasites (P=0.001, Student's t-test). However, incubation with heat-killed parasites or T. gondii lysates did not abrogate act D-induced apoptosis. In conclusion, inhibition of apoptosis by living, but not necessarily replicating T. gondii may facilitate parasite survival and persistence within its host cell. PMID- 10363678 TI - Osp17, a novel immunodominant outer surface protein of Borrelia afzelii: recombinant expression in Escherichia coli and its use as a diagnostic antigen for serodiagnosis of Lyme borreliosis. AB - Western blot analyses of the human humoral response of patients with Lyme borreliosis have shown that a 17-kDa protein is an immunodominant protein in late disease. Immune electron microscopy with a monoclonal antibody against this protein revealed that the 17-kDa protein is abundantly expressed on the surface of Borrelia afzelii strain PKo. Therefore, the protein has been renamed outer surface protein (Osp) 17. Recombinant Osp 17 of strain PKo was expressed in Escherichia coli and purified by chromatography. Immunoblot analysis of human sera showed a comparable sensitivity with recombinant and natural proteins. The DNA sequences of the osp17 genes from different B. afzelii strains were determined. The DNA sequences of the different osp 17 homologues (six isolates from skin, three isolates from CSF and one isolate from synovial fluid) had high sequence identities of at least 94%. Using a polyclonal antibody against recombinant Osp 17, it was shown that Osp 17 expression varied considerably among the investigated B. afzelii strains. As previously also observed for OspA- and OspC-encoding genes, the osp 17 gene is present in strains not expressing the respective protein. It has been shown that OspA and OspC expression varies in different environments such as tick and vertebrate host. Studies are underway to examine whether this is also true for Osp 17. For diagnostic purposes the use of recombinant Osp 17 has the advantage that the amount of Osp 17 antigen can be easily standardized for immunoblotting, and that this antigen can be used in a protein-specific enzyme-linked immunosorbent assay. PMID- 10363680 TI - Immune response to a recombinant fragment of the CagA protein of Helicobacter pylori in blood donors and patients with gastric cancer: relation to ABO(H) blood group phenotype, stage of the disease and tumor morphology. AB - IgG immune response to CagA was evaluated by enzyme-linked imunosorbent assay (ELISA) using a recombinant fragment of CagA as antigen in 171 patients with gastric cancer and 298 blood donors to determine whether it could be related to the ABO(H) blood group phenotype, stage of cancer or tumor morphology. The CagA ELISA showed a good specificity (93.5%) and sensitivity (88.5%) as compared with immunoblotting for blot CagA-negative and -positive donors. The Helicobacter pylori seropositive blood group A donors revealed the lowest proportion (37.6%) of strong responders to CagA: A3.0 was defined as significant intimal hyperplasia. RESULTS: Only 17 patients (30%) showed both a normal dobutamine stress echocardiogram and absence of significant intimal hyperplasia by intravascularultrasound. Abnormal findings were observed in 39 patients (70%): both by dobutamine stress echocardiography and intravascular ultrasound in 22 patients, by intravascular ultrasound alone in 11 patients, and by dobutamine stress echocardiography alone in 6 patients. Coronary flow velocity reserve did not discriminate between patients with normal or abnormal intravascular ultrasound or dobutamine stress echocardiographic findings. CONCLUSIONS: Only a minority of heart transplant patients with a normal coronary angiogram is free of pathological changes, when assessed by intravascular ultrasound and dobutamine stress echocardiography. Coronary flow velocity reserve does not seem useful to further characterize these patients. PMID- 10363682 TI - Arterial baroreflex modulation of heart rate in patients early after heart transplantation: lack of parasympathetic reinnervation. AB - BACKGROUND: Orthotopic heart transplantation results in cardiac denervation. The presence of cardiac parasympathetic reinnervation in humans has been widely debated based on the application of differing indirect measures of autonomic control. However no attempt has been made to analyse the reflex heart rate response to baroreceptor stimulation whose occurrence is generally considered a reliable marker of the ability to activate cardiac vagal reflexes. This study tested the hypothesis that the presence of donor heart RR interval lengthening following phenylephrine induced blood pressure increase would be an index of parasympathetic reinnervation. METHODS: Baroreflex sensitivity (BRS) was assessed in 30 patients (mean age 51+/-12 years) 1-24 months after heart transplantation carried out by the standard Lower-Shumway technique. In 6 patients the recipient atrium rate response (P-P interval) to baroreceptor stimulation by phenylephrine was also simultaneously determined by transesophageal recording. RESULTS: None of the 30 patients showed prolongation of RR intervals in the donor heart. The average BRS value was -0.28+/-0.54 ms/mmHg (range -1.3-0.7 ms/mm Hg). In the 6 patients in whom BRS was obtained at both the recipient atrium (P-P) and donor heart (R-R) the changes were 7.6+/-5.7 ms/mm Hg and -0.38+/-0.58 ms/mm Hg respectively (p = 0.02), thus confirming that the absent RR interval lengthening in the donor heart is the consequence of efferent vagal fiber interruption. CONCLUSIONS: The absence of any RR interval prolongation following phenylephrine induced baroreceptor stimulation demonstrates that vagal efferent reinnervation of the donor heart does not occur up to 24 months in patients operated via the standard Lower-Shumway procedure. It is also suggested that analysis of baroreceptor reflexes is a more specific method in the examination of cardiac parasympathetic reinnervation. PMID- 10363683 TI - A prospective randomized trial of complete atrioventricular transplantation versus ventricular transplantation with atrioplasty. AB - BACKGROUND: The standard technique of ventricular transplantation with atrioplasty (SOHT) distorts atrial anatomy. This may compromise diastolic ventricular function, impair atrioventricular valve competence and elevate resting ANP secretion. In contrast, complete atrioventricular anastomosis (CAVT) preserves atrial geometry. METHODS: We evaluated long term outcome in a prospective randomized trial of CAVT vs. SOHT. The primary outcome measures were peak oxygen uptake, atrioventricular valve regurgitation and ANP secretion. RESULTS: 58 recipients (median age 49 years; range 21-64) were consecutively randomized (29 CAVT; 29 SOHT). There were no differences in total ischaemic time, cardiopulmonary bypass time, postoperative bleeding or immunosuppression. Cardiopulmonary exercise tolerance testing was performed by 29 recipients at 742 to 1825 days. Pulmonary function was equivalent. Peak oxygen consumption expressed as a percentage of predicted maximum was 53.5% with CAVT and 63.8% with SOHT (p = 0.14). Echocardiography was performed on 41 recipients at 944 to 1665 days. There was less tricuspid regurgitation with CAVT (3/22 [13.6%] CAVT vs. 10/19 [52.6%] SOHT; p = 0.019). The incidence of mitral regurgitation was similar (5/22 [22.7%] CAVT vs. 4/19 [21.1%] SOHT; p = 0.803). Resting ANP secretion was assessed in 17 recipients at 1013 to 1812 days. All were hemodynamically stable and none had concurrent rejection. Resting ANP secretion was less with CAVT (CAVT: 283 pg/ml; SOHT: 521.4; p = 0.041). CONCLUSIONS: Peak oxygen consumption was not influenced by implantation technique. However, CAVT reduced the incidence of tricuspid regurgitation and attenuated the elevation in resting ANP secretion. PMID- 10363684 TI - Determinants of waiting time for heart transplants in the United States. AB - BACKGROUND: Reports have been published on factors affecting the variations in waiting times for kidney and liver transplant candidates who have been registered on the United Network for Organ Sharing's waiting list. This study reports on determinants of waiting time differences that occur in the eleven UNOS regions for heart transplant candidates. METHODS: Retrospective analysis of 11,345 primary heart waiting-list registrations and 15,868 cadaveric donors, from whom 7,043 hearts were recovered and transplanted for the years 1994-96. Because estimated populations in the eleven UNOS regions vary from 10.8 to 43.2 million, analyses utilized Registrations/million population and Transplants/million population to obtain an R/T ratio. The relationship of the R/T ratio to the median waiting time was then examined for different demographic variables. RESULTS: The numbers of new heart candidate registrations, heart transplants performed, and waiting list deaths have undergone little change from 1991 through 1996. National median waiting times varied by basic demographic variables such as ABO blood type, race, age group, and UNOS medical urgency status. In the eleven UNOS regions, registrations per million ranged from 11.5 to 33.0 and transplants per million from 5.3 to 10.7. Registration/Transplant ratios correlated with median waiting times for urgency Status 1 and 2 as well as for blood group O recipients. Correlation with blood type AB recipients was less consistent, in part, due to the small number of AB recipients. CONCLUSIONS: There are wide variations in the number of heart transplant candidate registrations and in the number of heart transplants performed in the eleven UNOS regions. The registration to transplantation ratio correlated with median waiting times in these regions. Factors possibly contributing to the observed variations were examined. PMID- 10363685 TI - Effect of immunosuppressive therapy, serum creatinine, and time after transplant on plasma total homocysteine in patients following heart transplantation. AB - OBJECTIVES: To determine the prevalence of hyperhomocysteinemia in heart transplant recipients, and to assess the effect of renal function and immunosuppressive medication on total plasma homocysteine (tHcy) levels. BACKGROUND: Elevated plasma tHcy levels have been associated with increased risk of mortality in patients with established coronary artery disease. Graft coronary disease is the major cause of morbidity and mortality in long-term survivors of heart transplantation. The tHcy has been found to be elevated in heart and kidney transplant patients, however, the etiologic factors have not been clearly delineated. METHODS: The study group consisted of 70 heart transplant recipients (56 males, 14 females, mean age 53+/-13 years [range 17 to 69 years]). The parameters evaluated were fasting tHcy level, cumulative cyclosporine (CyA) dose, cumulative prednisone dose, serum creatinine, and time from transplantation. RESULTS: The mean fasting tHcy level was 20.5+/-10.2 micromol/L (range 5.2 to 59.0 micromol/L). Sixty-one (87%) had fasting tHcy levels greater than the seventy-fifth percentile of the general population (>12.2 micromol/L in males, and >10.1 micromol/L in females). There was no difference in mean post-transplant tHcy level between patients with and without coronary artery disease before transplantation (21.0+/-11.4 vs. 19.3+/-6.7 micromol/L, p = NS). There were significant relationships between the tHcy level and the serum creatinine (r = 0.76, p<0.001), and cumulative exposure to CyA (r = 0.31, p<0.01). There were no significant relationships between tHcy levels and cumulative prednisone dose, or time from transplantation. CONCLUSIONS: Fasting tHcy levels are markedly elevated in the majority of patients following heart transplantation, and are correlated to serum creatinine. Further studies are needed to determine other etiologic factors of elevated tHcy following heart transplantation, and to examine the impact of elevated tHcy on clinical outcomes. PMID- 10363686 TI - Influence of graft ischemic time and donor age on survival after lung transplantation. AB - BACKGROUND: Increased graft ischemic time and donor age are risk factors for early death after heart transplantation, but the effect of these variables on survival after lung transplantation has not been determined in a large, multinational study. METHODS: All recipients of cadaveric lung transplantations performed between October 1, 1987 and June 30, 1997 which were reported to the United Network for Organ Sharing/International Society for Heart and Lung Transplantation (UNOS/ISHLT) Registry were analyzed. Patient survival rates were estimated using Kaplan-Meier methods. Multivariate logistic regression was used to determine the impact of donor and recipient characteristics on patient survival after transplantation. To examine whether the impact of donor age varied with ischemic time, interactions between the 2 terms were examined in a separate multivariate logistic regression model. RESULTS: Kaplan-Meier survival did not differ according to the total lung graft ischemia time, but recipient survival was significantly adversely affected by young (-10 years) or old (-51 years) donor age (p = 0.01). On multivariate analysis, neither donor age nor lung graft ischemic time per se were independent predictors of early survival after transplantation, except if quadratic terms of these variables were included in the model. The interaction between donor age and graft ischemia time, however, predicted 1 year mortality after lung transplantation (p = 0.005), especially if donor age was greater than 55 years and ischemic time was greater than 6 to 7 hours. CONCLUSIONS: Graft ischemia time alone is not a risk factor for early death after lung transplantation. Very young or old donor age was associated with decreased early survival, whereas the interaction between donor age and ischemic time was a significant predictor of 1 year mortality after transplantation. Cautious expansion of donor acceptance criteria (especially as regards ischemic time) is advisable, given the critical shortage of donor lung grafts. PMID- 10363687 TI - Comparison between mycophenolate mofetil- and azathioprine-based immunosuppressions in clinical lung transplantation. AB - BACKGROUND: The aim of the study was to assess the impact of mycophenolate mofetil (MMF) on the early phase after lung transplantation. PATIENTS AND METHODS: Thirty-eight consecutive patients between November 1994 and January 1997 were treated with cyclosporine, prednisolone, antithymocyte globuline induction therapy, and either MMF (n = 21) or azathioprine (Aza) (n = 17). Four patients from the MMF group and 2 patients from the Aza group were intubated and in the ICU prior to transplantation. Demographic data and primary diagnosis were comparable. MMF was administered at a dosage of 2 gm/day whereas Aza was initiated at 2 mg/kg/day and adapted by leukocyte count. Three-month survival and incidence of rejections and infections were compared. RESULTS: Six-month survival in the MMF group was 76% compared to 65% in the Aza group (n.s.). The mean number of acute rejection episodes in the MMF and Aza group were 0.29+/-0.10 and 1.53+/ 0.29 (p<0.01) respectively. Transbronchial biopsy (TBB) results > or =grade 2 ISHLT were seen in 10% of MMF and in 43% of Aza-treated patients; completely free from rejection were 17 MMF and 3 Aza patients. The mean number of infections per patient in the MMF and Aza group were 1.57+/-0.29 and 2.29+/-0.40 respectively, bacterial (1.10 vs. 1.71), viral (0.35 vs. 0.33), and fungal (0.14 vs. 0.24) infections were the same in both groups. CONCLUSIONS: These data result suggest that mycophenolate mofetil therapy is more effective in preventing rejection episodes in patients early after lung transplantation than therapy with azathioprine. We therefore conclude that MMF is a safe and effective drug to optimize immunosuppressive therapy in the early phase after lung transplantation. PMID- 10363688 TI - Cyclosporine inhibits long-term survival in cardiac allografts treated with monoclonal antibody against CD45RB. AB - BACKGROUND: We have previously reported that a monoclonal antibody to CD45RB is a novel immunosuppressive agent; however, the optimal regimen in cardiac allografts remains unknown. The present study was undertaken to determine the optimal protocol of this therapy and its interaction with cyclosporine. METHODS: A heterotopic heart allograft model was used in C57BL/6 to BALB/c mice. The following studies were conducted: 1) dose response study (low, intermediate, and high doses at 1, 3, and 9 mg/kg/day respectively), 2) short course (2 days) therapy vs. long course (9 days) therapy, 3) pretreatment (starting on day -1) vs no pretreatment, 4) daily therapy vs. alternative day therapy, and 5) monoclonal antibody treatment with and without cyclosporine. RESULTS: The efficacy of the CD45RB monoclonal antibody was dose and duration dependent (p<0.01). Pretreatment significantly improved the efficacy of this therapy (74.5+/-13.4 days vs. 30.6+/ 1.5 days, p<0.01). Daily therapy was superior to alternate day therapy (74.5+/ 13.4 days vs. 30.4+/-1.5 days, p<0.03). Interestingly, we found that administration of cyclosporine prior to, at the same time as, or after administration of the CD45RB monoclonal antibody had a detrimental effect on graft survival compared to mAb treated alone (16.6+/-0.4 days, 25+/-2.3 days, and 35.3+/-0.9 days respectively vs. 74.5 days, p<0.01). CONCLUSIONS: Immunosuppression with CD45RB monoclonal antibody is dose and duration dependent. Pretreatment and daily therapy improves results. Addition of cyclosporine inhibits long-term graft survival achieved by the monoclonal antibody alone. PMID- 10363689 TI - Efficacy of tacrolimus in the treatment of refractory rejection in heart and lung transplant recipients. AB - BACKGROUND: Refractory acute cellular rejection may occur despite triple-drug immunosuppression (cyclosporine A, steroids, azathioprine/mycophenolate mofetil). The purpose of this study was to determine the efficacy of tacrolimus rescue therapy in patients maintained on cyclosporine-based immunosuppression (CBI). METHODS: Between December 1993 and October 1996, 208 patients underwent thoracic organ transplantation at the Hospital of the University of Wisconsin at Madison. One hundred forty-nine patients underwent heart replacement; 59 underwent lung transplantation. One hundred thirty-nine of the heart transplant cohort received CBI preceded by induction therapy with OKT3. Forty-six of the lung transplant cohort received CBI without induction cytolytic therapy. Refractory rejection was defined as failure to respond to high-dose steroids (500 mg to 1 g IV methylprednisolone for 3 days) and/or monoclonal antibody therapy (OKT3, 5 to 10 mg IV/day for 7 to 14 days). In patients with refractory rejection, cyclosporine was replaced with tacrolimus. RESULTS: Overall, 16% (30/185) of patients receiving CBI experienced refractory rejection. Thirty-one episodes of grade IIIa or greater rejection occurred in 11% (15/139) of heart transplant recipients. Twenty episodes of grade II to IV rejection occurred in 33% (15/46) of lung transplant recipients. After tacrolimus rescue therapy, 93% (14/15) of patients in the heart transplant group converted to grade II or less rejection. Refractory rejection was reversed in 73% (11/15) of the lung transplant group. Reversal was documented at biopsy in all (8/8) lung recipients in whom it had been histologically identified. FEV1 values of 3 additional patients stabilized. CONCLUSIONS: The incidence of refractory rejection in thoracic organ transplant recipients on CBI is significant. Reversal of refractory rejection follows rescue immunotherapy with tacrolimus. PMID- 10363690 TI - Hepatitis C virus infection and lung transplantation: a survey of practices. AB - BACKGROUND: Approximately 4 million persons in the United States are chronically infected with hepatitis C and morbidity due to this disease is increasingly observed in transplant recipients. While knowledge of hepatitis C in liver and kidney transplantation is advancing, little information is available concerning hepatitis C and lung transplantation. We surveyed lung transplant programs about policies regarding testing for hepatitis C, transplantation of hepatitis C infected candidates, and the use of organs from seropositive donors. METHODS: A written questionnaire was sent to all United Network of Organ Sharing (UNOS) approved lung transplant programs. RESULTS: Fifty-nine of 89 (66%) surveys were returned, including 49 from active programs, capturing 81% of lung transplants performed within UNOS prior to January 1998. All programs screen candidates for hepatitis C. The estimated median seropositivity rate among candidates was 1.9%. Thirty-three of 46 (72%) programs consider seropositive patients for transplantation and most use virologic and/or histologic data to determine candidacy. All donors are screened for hepatitis C. Twenty-six of 47 (55%) programs accept lungs from seropositive donors and many restrict the use of organs from seropositive donors to infected recipients. Few programs routinely test recipients for hepatitis C, and policies for monitoring those with known infection are variable. CONCLUSIONS: Lung transplant candidates and donors are tested routinely for hepatitis C. The majority of programs are willing to accept infected candidates and seropositive donors. Post-transplant follow-up of hepatitis C is variable and prospective studies are needed to evaluate the impact of hepatitis C on lung transplant recipients. PMID- 10363691 TI - Induction immunotherapy in pediatric heart transplant recipients: a multicenter study. AB - BACKGROUND: The efficacy and safety of induction immunotherapy with antithymocyte antibody preparations (IND) in pediatric heart transplantation is controversial. Experimentally, recipient age is an important determinant of immune responses. The effects on induction immunotherapy were determined by an analysis of outcomes of 465 pediatric (age <18 years) heart recipients that either did or did not receive IND in the first week post-transplant. METHODS: The outcomes of 2 groups who received either OKT3 (n = 101) or rabbit polyclonal antithymocyte serum (N/R ATS, n = 105) were compared with 255 recipients who did not receive antithymocyte antibodies. The study population were all heart recipients enrolled in the Pediatric Heart Transplant Study Group (PHTS) between January 1993 and December 1995 and followed up to 36 months. RESULTS: Overall mortality and death due to rejection were lowest with N/R-ATS IND (8/105 and 1/105, respectively) compared with the no-induction group (58/255 and 8/ 255, respectively) or the OKT3 group (22/101 and 7/101, respectively) with significance of p = 0.001 and 0.06 respectively. Late mortality beyond 30 days after transplant was lowest with N/R ATS IND compared with the OKT3 and no IND (p = 0.01). Induction did not affect cumulative infections, deaths due to infection, or the frequency of malignancies. Patients excluded from N/R-ATS induction had the highest mortality (18/ 43), suggesting that the protocol's exclusion criteria identified a high-risk group. To minimize potential effect(s) of exclusion bias, patients transplanted at centers participating in the N/R-ATS induction trial were reanalyzed with a post hoc intent-to-treat analysis assigning patients by center (IND or no IND) irrespective of actual treatment. With this analysis overall mortality was 18% for N/R-ATS centers, 21% for OKT3 centers, and 26% for centers not using IND (p = 0.3). The mortalities of recipients <6 months old at transplant were lowest at centers using N/R-ATS and OKT3 IND compared to centers not using IND (p = 0.04). Cumulative rejection (0.8 vs 1.2 rejection/pt/year, p = 0.01) and freedom from rejection death (99% vs. 93% at year 1, p = 0.02) of the N/R-ATS centers were lower compared to OKT3 centers but were not different from centers not using IND. CONCLUSION: Following orthotopic transplantation, induction immunotherapy can exert the enduring benefit of reducing late deaths, a possible surrogate for rejection severity, in recipients less than 6 months of age, while not being associated with higher rates of infectious or malignant complications. Since polyclonal anti-T cell antibody preparations appears superior to OKT3 induction in pediatric recipients, the efficacy of ATS induction should be further evaluated in a randomized prospective study in pediatric heart recipients. PMID- 10363692 TI - Captopril and platelet-activating factor (PAF) antagonist prevent cardiac allograft vasculopathy in rats: role of endogenous PAF and PAF-like compounds. AB - Accelerated coronary artery disease (CAD) is the leading cause of late mortality following cardiac transplantation. The vascular lesions are characterized by myointimal proliferation and perivascular mononuclear inflammatory infiltrates. Platelet-activating factor (PAF, 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) is a potent phospholipid mediator produced by inflammatory cells and activated endothelial cells. Angiotensin II is known to activate phospholipase A2, a critical enzyme in PAF synthesis. Using a rat heterotopic cardiac transplant model known to induce graft CAD, we previously reported that chronic administration of captopril, an angiotensin converting enzyme inhibitor, reduces intimal proliferation and maintains luminal patency. The purpose of the current study was to determine if captopril regulates vascular remodeling by suppressing PAF synthesis and whether administration of a PAF antagonist ameliorates graft CAD. Captopril was found to decrease levels of PAF and PAF-like compounds as well as reduce intimal lesions, decrease cellular rejection grade, and diminish allograft heart weights. Treatment with a PAF antagonist significantly decreased proliferation of the intimal component of the vasculopathy and caused regression of the cardiac hypertrophy, but had no significant effect on cellular rejection. In contrast, untreated animals had elevated plasma PAF levels, elevated heart weights, and severe myointimal proliferation with luminal stenosis 21 days post transplantation. These observations suggest that graft CAD is mediated, in part, by PAF and PAF-like compounds, and suppression of endogenous PAF may prevent cardiac allograft vasculopathy. PMID- 10363694 TI - Age before beauty: the use of "older" donor hearts for cardiac transplantation. PMID- 10363693 TI - Changes in oxidative stress and cellular redox potential during myocardial storage for transplantation: experimental studies. AB - BACKGROUND: Cardioplegic solutions assure only a sub-optimal myocardial protection during prolonged storage for transplantation. The ultimate cause of myocardial damage during storage is unknown, but oxygen free radicals might be involved. We evaluated the occurrence of oxidative stress and changes in cellular redox potential after different periods of hypothermic storage. METHODS: Langendorff-perfused rabbit hearts were subjected to a protocol mimicking each stage of a cardiac transplantation procedure: explantation, storage and reperfusion. Three periods of storage were considered: Group A = 5 hours, Group B = 15 hours, and Group C = 24 hours. Oxidative stress was determined in terms of myocardial content and release of reduced (GSH) and oxidized (GSSG) glutathione, and cellular redox potential as oxidized and reduced pyridine nucleotides ratio (NAD/NADH). Data on mechanical function, cellular integrity and myocardial energetic status were collected. RESULTS: At the end of reperfusion, despite the different timings of storage, recovery of left ventricular developed pressure (46.1+/-7.0, 54.7+/-6.7, and 45.7+/-7.4% of the baseline pre-ischaemic value), energy charge (0.81+/-0.02, 0.81+/-0.02, and 0.77+/-0.01) and NAD/NADH ratio (8.87+/-1.08, 9.39+/-1.72, and 10.26+/-1.98) were similar in all groups (A, B and C). On the contrary, the rise in left ventricular resting pressure (LVRP) and GSH/GSSG ratio were significantly different between Group C, and Groups A and B (p<0.0001, analyzed by Generalized Estimating Equations model for repeated measures, and p<0.05, respectively). CONCLUSIONS: The pathophysiology of myocardial damage during hypothermic storage cannot be considered as a normothermic ischaemic injury and parameters other than energetic metabolism, such as thiolic redox state, are more predictive of functional recovery upon reperfusion. PMID- 10363695 TI - Cyclosphosphamide/prednisone for combination immunosuppression and therapy of lymphoproliferative disease. AB - Post-transplant lymphoproliferative disease (PTLD) is a well-known complication of solid organ transplantation. While this disorder can often be controlled by decreasing immunosuppression, it is frequently difficult to control the resultant rejection. This case exemplifies this dilemma. To solve this problem, cyclosphosphamide (600 mg/m2) and prednisone (2 mg/kg times 5 days) were given every 3 weeks to simultaneously treat PTLD and provide immunosuppression. PMID- 10363696 TI - Prospective cost-utility analysis of lung transplantation in The Netherlands: a comment on the pilot study by Gartner et al. PMID- 10363697 TI - Fast spatio-temporal image reconstruction for dynamic PET. PMID- 10363698 TI - Iterative reconstruction of single-shot spiral MRI with off resonance. AB - A variety of applications and research directions in magnetic resonance imaging which require fast scan times have recently become popular. In order to satisfy many of the requirements of these applications, snapshot imaging methods, which acquire an entire image in one excitation, are often used. These snapshot techniques are relatively insensitive to motion and can allow rapidly occurring processes to be imaged. However, snapshot imaging techniques acquire data over a relatively long period, during which off-resonance phase can accumulate, leading to image degradation. This degradation often limits the usefulness of the images. Presented here is a method to iteratively reconstruct an image acquired by a spiral snapshot technique and to remove image degradation due to off resonance. This iterative method does not assume that the inhomogeneity is slowly varying within the image, allowing better results than with deblurring techniques which do not take abrupt changes into account. Although presented here with a spiral imaging technique, the iterative algorithm is general enough to be applied to a variety of snapshot imaging techniques. PMID- 10363699 TI - Automated extraction and variability analysis of sulcal neuroanatomy. AB - Systematic mapping of the variability in cortical sulcal anatomy is an area of increasing interest which presents numerous methodological challenges. To address these issues, we have implemented sulcal extraction and assisted labeling (SEAL) to automatically extract the two-dimensional (2-D) surface ribbons that represent the median axis of cerebral sulci and to neuroanatomically label these entities. To encode the extracted three-dimensional (3-D) cortical sulcal schematic topography (CSST) we define a relational graph structure composed of two main features: vertices (representing sulci) and arcs (representing the relationships between sulci). Vertices contain a parametric representation of the surface ribbon buried within the sulcus. Points on this surface are expressed in stereotaxic coordinates (i.e., with respect to a standardized brain coordinate system). For each of these vertices, we store length, depth, and orientation as well as anatomical attributes (e.g., hemisphere, lobe, sulcus type, etc.). Each arc stores the 3-D location of the junction between sulci as well as a list of its connecting sulci. Sulcal labeling is performed semiautomatically by selecting a sulcal entity in the CSST and selecting from a menu of candidate sulcus names. In order to help the user in the labeling task, the menu is restricted to the most likely candidates by using priors for the expected sulcal spatial distribution. These priors, i.e., sulcal probabilistic maps, were created from the spatial distribution of 34 sulci traced manually on 36 different subjects. Given these spatial probability maps, the user is provided with the likelihood that the selected entity belongs to a particular sulcus. The cortical structure representation obtained by SEAL is suitable to extract statistical information about both the spatial and the structural composition of the cerebral cortical topography. This methodology allows for the iterative construction of a successively more complete statistical models of the cerebral topography containing spatial distributions of the most important structures, their morphometrics, and their structural components. PMID- 10363700 TI - Anatomical model matching with fuzzy implicit surfaces for segmentation of thoracic volume scans. AB - Many segmentation methods for thoracic volume data require manual input in the form of a seed point, initial contour, volume of interest etc. The aim of the work presented here is to further automate this segmentation initialization step. In this paper an anatomical modeling and matching method is proposed to coarsely segment thoracic volume data into anatomically labeled regions. An anatomical model of the thorax is constructed in two steps: 1) individual organs are modeled with blended fuzzy implicit surfaces and 2) the single organ models are grouped into a tree structure with a solid modeling technique named constructive solid geometry (CSG). The combination of CSG with fuzzy implicit surfaces allows a hierarchical scene description by means of a boundary model, which characterizes the scene volume as a boundary potential function. From this boundary potential, an energy function is defined which is minimal when the model is registered to the tissue-air transitions in thoracic magnetic resonance imaging (MRI) data. This allows automatic registration in three steps: feature detection, initial positioning and energy minimization. The model matching has been validated in phantom simulations and on 15 clinical thoracic volume scans from different subjects. In 13 of these sets the matching method accurately partitioned the image volumes into a set of volumes of interest for the heart, lungs, cardiac ventricles, and thorax outlines. The method is applicable to segmentation of various types of thoracic MR-images, provided that a large part of the thorax is contained in the image volume. PMID- 10363701 TI - Statistical textural features for detection of microcalcifications in digitized mammograms. AB - Clustered microcalcifications on X-ray mammograms are an important sign for early detection of breast cancer. Texture-analysis methods can be applied to detect clustered microcalcifications in digitized mammograms. In this paper, a comparative study of texture-analysis methods is performed for the surrounding region-dependence method, which has been proposed by the authors, and conventional texture-analysis methods, such as the spatial gray-level dependence method, the gray-level run-length method, and the gray-level difference method. Textural features extracted by these methods are exploited to classify regions of interest (ROI's) into positive ROI's containing clustered microcalcifications and negative ROI's containing normal tissues. A three-layer backpropagation neural network is used as a classifier. The results of the neural network for the texture-analysis methods are evaluated by using a receiver operating characteristics (ROC) analysis. The surrounding region-dependence method is shown to be superior to the conventional texture-analysis methods with respect to classification accuracy and computational complexity. PMID- 10363702 TI - Pixelwise fusion for optimizing SNR in multiple-plate computed radiography imaging. AB - The computed radiography (CR) technique, also known as the storage phosphor imaging technique, has evolved to be a major candidate for large-scale implementation of digital radiography during the past decade. In order to obtain a reasonable spatial resolution, the storage phosphor plate used is generally limited in thickness. This leads to X rays being only partially absorbed by the detector. Useful information may be contained in the X rays transmitted through the detector. Multiple-plate imaging techniques may be used to capture and utilize the X rays more efficiently. In this paper, an image fusion method, based on the Rayleigh principle and the Karhunen-Loeve (K-L) transform, is presented for optimizing the signal-to-noise ratio (SNR) of the fused image on a pixel-by pixel basis. Because the multiple-plate images contain the same structural information, the signal components of the images are highly correlated with one another. Thus, the K-L transform is applied to decompose each of the multiple plate images into an eigen image (the estimated signal) and a residual image (the estimated noise). An average representation entropy measure is maximized for selecting the number of eigen components to be included in the signal estimation. An experimental study, using an anthropomorphic chest phantom, is presented to illustrate pixelwise fusion of multiple-plate images. Experimental results show that the SNR of the fused image was improved by 12-48%, depending upon the anatomical regions of interest in the image. PMID- 10363703 TI - An EM algorithm for dynamic SPECT. AB - In this paper we present two variants of the EM algorithm for dynamic SPECT imaging. A version based on compartmental modeling which fits a sum of exponentials and a more general approach allowing for arbitrary decaying activities. The underlying probabilistic models are discussed and the incomplete and complete data spaces are shown to be physically meaningful. We indicate that the second method, leading to a convex program in the M step, is easier to treat numerically and we present a possible numerical approach. Some preliminary numerical tests indicating the feasibility of the method are included. PMID- 10363704 TI - Gradient-based iterative image reconstruction scheme for time-resolved optical tomography. AB - Currently available tomographic image reconstruction schemes for optical tomography (OT) are mostly based on the limiting assumptions of small perturbations and a priori knowledge of the optical properties of a reference medium. Furthermore, these algorithms usually require the inversion of large, full, ill-conditioned Jacobian matrixes. In this work a gradient-based iterative image reconstruction (GIIR) method is presented that promises to overcome current limitations. The code consists of three major parts: 1) A finite-difference, time resolved, diffusion forward model is used to predict detector readings based on the spatial distribution of optical properties; 2) An objective function that describes the difference between predicted and measured data; 3) An updating method that uses the gradient of the objective function in a line minimization scheme to provide subsequent guesses of the spatial distribution of the optical properties for the forward model. The reconstruction of these properties is completed, once a minimum of this objective function is found. After a presentation of the mathematical background, two- and three-dimensional reconstruction of simple heterogeneous media as well as the clinically relevant example of ventricular bleeding in the brain are discussed. Numerical studies suggest that intraventricular hemorrhages can be detected using the GIIR technique, even in the presence of a heterogeneous background. PMID- 10363705 TI - Wavelet-based lossless compression of coronary angiographic images. AB - The final diagnosis in coronary angiography has to be performed on a large set of original images. Therefore, lossless compression schemes play a key role in medical database management and telediagnosis applications. This paper proposes a wavelet-based compression scheme that is able to operate in the lossless mode. The quantization module implements a new way of coding of the wavelet coefficients that is more effective than the classical zerotree coding. The experimental results obtained on a set of 20 angiograms show that the algorithm outperforms the embedded zerotree coder, combined with the integer wavelet transform, by 0.38 bpp, the set partitioning coder by 0.21 bpp, and the lossless JPEG coder by 0.71 bpp. The scheme is a good candidate for radiological applications such as teleradiology and picture archiving and communications systems (PACS's). PMID- 10363706 TI - Tracking the left ventricle in echocardiographic images by learning heart dynamics. AB - In this paper a temporal learning-filtering procedure is applied to refine the left ventricle (LV) boundary detected by an active-contour model. Instead of making prior assumptions about the LV shape or its motion, this information is incrementally gathered directly from the images and is exploited to achieve more coherent segmentation. A Hough transform technique is used to find an initial approximation of the object boundary at the first frame of the sequence. Then, an active-contour model is used in a coarse-to-fine framework, for the estimation of a noisy LV boundary. The PCA transform is applied to form a reduced ordered orthonormal basis of the LV deformations based on a sequence of noisy boundary observations. Then this basis is used to constrain the motion of the active contour in subsequent frames, and thus provide more coherent identification. Results of epicardial boundary identification in B-mode images are presented. PMID- 10363707 TI - Temporal tuning of odor responses in pheromone-responsive projection neurons in the brain of the sphinx moth Manduca sexta. AB - By means ofintracellular recording and staining, we studied the ability of several distinct classes of projection (output) neurons, which innervate the sexually dimorphic macroglomerular complex (MGC-PNs) in the antennal lobe of the male moth Manduca sexta, to encode naturally intermittent sex pheromonal stimuli. In many MGC-PNs, antennal stimulation with a blend of the two essential pheromone components evoked a characteristic triphasic response consisting of a brief, hyperpolarizing inhibitory potential (I1) followed by depolarization with firing of action potentials and then a delayed period of hyperpolarization (I2). MGC-PNs described in this study resolved pulsed pheromonal stimuli, up to about five pulses/second, with a distinct burst of action potentials for each pulse of odor. The larger the amplitude of I1, the higher the pulse rate an MGC-PN could follow, illustrating the importance of inhibitory synaptic input in shaping the temporal firing properties of these glomerular output neurons. In some MGC-PNs, triphasic responses were evoked by antennal stimulation with only one of the two key pheromone components. Again, the maximal pulse rate that an MGC-PN could follow with that pheromone component as sole stimulus was high in MGC-PNs that responded with a strong I1. These component-specific MGC-PNs innervated only one of the two principal glomeruli of the MGC, while MGC-PNs that were primarily excited by antennal stimulation with either key pheromone component had arborizations in both major MGC glomeruli. These observations therefore suggest that the population of antennal olfactory receptor cells responding to a single pheromone component is functionally heterogeneous: a subset of these sensory cells activates the excitatory drive to many uniglomerular MGC-PNs, while others feed onto inhibitory circuits that hyperpolarize the same PNs. This convergence of opposing inputs is a circuit property common to uniglomerular MGC-PNs branching in either of the major MGC glomeruli, and it enhances the ability of these glomerular output neurons to resolve intermittent olfactory input. Synaptic integration at the uniglomerular PN level thus contributes to the transmission of behaviorally important temporal information about each key pheromone component to higher centers in the brain. PMID- 10363708 TI - Alterations in size, number, and morphology of gustatory papillae and taste buds in BDNF null mutant mice demonstrate neural dependence of developing taste organs. AB - Sensory ganglia that innervate taste buds and gustatory papillae (geniculate and petrosal) are reduced in volume by about 40% in mice with a targeted deletion of the gene for brain-derived neurotrophic factor (BDNF). In contrast, the trigeminal ganglion, which innervates papillae but not taste buds on the anterior tongue, is reduced by only about 18%. These specific alterations in ganglia that innervate taste organs make possible a test for roles of lingual innervation in the development of appropriate number, morphology, and spatial pattern of fungiform and circumvallate papillae and associated taste buds. We studied tongues of BDNF null mutant and wild-type littermates and made quantitative analyses of all fungiform papillae on the anterior tongue, the single circumvallate papilla on the posterior tongue, and all taste buds in both papilla types. Fungiform papillae and taste buds were reduced in number by about 60% and were substantially smaller in diameter in mutant mice 15-25 days postnatal. Remaining fungiform papillae were selectively concentrated in the tongue tip region. The circumvallate papilla was reduced in diameter and length by about 40%, and papilla morphology was disrupted. Taste bud number in the circumvallate was reduced by about 70% in mutant tongues, and the remaining taste buds were smaller than those on wild-type tongues. Our results demonstrate a selective dependence of taste organs on a full complement of appropriate innervation for normal growth and morphogenesis. Effects on papillae are not random but are more pronounced in specific lingual regions. Although the geniculate and petrosal ganglia sustain at least half of their normal complement of cell number in BDNF /- mice, remaining ganglion cells do not substitute for lost neurons to rescue taste organs at control numbers. Whereas gustatory ganglia and the taste papillae initially form independently, our results suggest interdependence in later development because ganglia derive BDNF support from target organs and papillae require sensory innervation for morphogenesis. PMID- 10363709 TI - Dopamine transporter-immunoreactive neurons decrease with age in the human substantia nigra. AB - Unbiased disector stereologic cell counting was applied to sections from the human substantia nigra that were immunostained by using a monoclonal antibody against the dopamine transporter (DAT). This antibody was found to penetrate the full thickness of the stained section. Quantification of the number of DAT immunostained neurons was performed in human cases stratified into three age groups, young (ages 0-49 years), middle aged (ages 50-69 years), and aged (ages 70-85 years). The number of DAT-immunoreactive nigral neurons was normalized for each case by constructing a ratio of the number of DAT-containing neurons to total number of neuromelanin-containing cells in each subject's sample. Three types of DAT nigral neurons were seen: type 1, intensely stained; type 2, lightly stained; and type 3, DAT-immunonegative neuromelanin-containing perikarya. By 50 years of age, the number of type 1 neurons decreased significantly (P < 0.0001), whereas the number of type 2 neurons increased with age (P < 0.0001). Type 3 neurons also increased with age (P < 0.01), although less robustly than type 2 neurons. Type 1 neurons decreased by 11.2% per decade, and the total number of nigral neurons (types 1-3) decreased by 6.7% per decade. Relative to the young group, there were 75% and 88% reductions in type 1 neurons in the middle-aged and aged groups, respectively. This contrasts with the 35% and 41% reductions in total number of neuromelanin-containing neurons seen in middle-aged and aged groups, respectively. The young group had significantly more type 1 neurons and fewer type 2 neurons compared with middle-aged and aged participants. Post-hoc analyses indicated that the young group had significantly fewer type 3 neurons compared with middle-aged and aged participants. These findings demonstrate an age-related reduction in the number of substantia nigra DAT-immunoreactive neurons. Therefore, insight into the mechanisms regulating the rate of DAT synthesis may aid in our understanding of the decline of DATs with aging and its functional significance. PMID- 10363710 TI - Immunocytochemical localization of the dopamine transporter in human brain. AB - The dopamine transporter (DAT) was localized in normal human brain tissue by light microscopic immunocytochemistry by using highly specific monoclonal antibodies. Regional distribution of DAT was found in areas with established dopaminergic circuitry, e.g., mesostriatal, mesolimbic, and mesocortical pathways. Mesencephalic DAT-immunoreactivity was enriched in the dendrites and cell bodies of neurons in the substantia nigra pars compacta and ventral tegmental area. Staining in the striatum and nucleus accumbens was dense and heterogeneous. Mesocortical DAT immunoreactivity in motor, premotor, anterior cingulate, prefrontal, entorhinal/perirhinal, insular, and visual cortices was detected in scattered varicose and a few nonvaricose fibers. Varicose fibers were relatively enriched in the basolateral and central subnuclei of amygdala, with sparser fibers in lateral and basomedial subnuclei. Double-labeling studies combining DAT and tyrosine hydroxylase (TH) immunostaining in the ventral mesencephalon showed two subpopulations of dopaminergic neurons differentiated by the presence or absence of DAT-immunoreactivity in the A9 and A10 cell groups. In other dopaminergic cell groups (All, A13-A15), TH-positive hypothalamic neurons showed no detectable DAT-immunoreactivity. However, fine DAT-immunoreactive axons were scattered throughout the hypothalamus, particularly concentrated along the medial border, with more coarse axons present along the lateral border. These findings demonstrate that most mesotelencephalic dopamine neurons of human brain express high levels of DAT throughout their entire somatodendritic and axonal domains, whereas a smaller subpopulation of mesencephalic dopamine cells and all hypothalamic dopamine cell groups examined express little or no DAT. These data indicate that different subpopulations of dopaminergic neurons use different mechanisms to regulate their extracellular dopamine levels. PMID- 10363711 TI - Laminar boundaries persist in the hippocampal dentate molecular layer of the mutant Shaking Rat Kawasaki despite aberrant granule cell migration. AB - The present report provides the first detailed description of the hippocampus in the Shaking Rat Kawasaki (SRK) mutant by using a panel of antibody markers to delineate its laminar organization. The mutant was characterised at postnatal day 21 by severe malformations of both neuronal position and orientation, the most striking of which was the presence of a rounded central granule cell mass in the dentate gyrus rather than the normal V-shaped granule cell layer. Despite this finding, the SRK dentate gyrus not only retained a cell-sparse molecular layer (thinner but similar in gross appearance to that of control littermates), but the sharp laminar boundary between its inner and outer parts was as clearly marked by IM1 and OM4 antibody staining as it was in the normal dentate gyrus. These immunocytochemical data suggest that the entorhinal terminal field of the dentate gyrus may be relatively normal in the mutant, despite entorhinal afferents appearing to take an abnormal trajectory after they fail to cross the hippocampal fissure. Laminar malformations included disruption of the SRK pyramidal cell layer, with spreading of the CA3 mossy fibre projection to an ectopic infrapyramidal position, radial displacement of CA1 pyramids, and transposition of a hitherto unremarked longitudinal fibre bundle immunoreactive for calretinin from its normal position in the stratum lacunosum-moleculare of field CA2 to an alvear position in SRK. The SRK malformations were very like but not identical to those seen in the mouse reeler mutant, suggesting similar underlying developmental mechanisms. PMID- 10363712 TI - Ultrastructural change at rat cerebellothalamic synapses associated with volitional motor adaptation. AB - Our ability to develop or modify motor skills is thought to involve persistent changes in the efficacy of synaptic transmission (synaptic plasticity) in the cerebellum. Previous work from our laboratory and others, examining synapses between neurons in the deep cerebellar nuclei and neurons in the thalamus revealed ultrastructural characteristics that have been implicated in the expression of synaptic plasticity at other locations in the brain. The present study sought evidence of ultrastructural plasticity at cerebellothalamic synapses associated with volitional motor adaptation. Adult rats were subject to 21 days of training, throughout which a novel load (overcome by predominantly shoulder adduction) was applied to the left forelimb while they fed (the right forelimb acted as an internal control). The behavioral paradigm was observed to produce a profound unilateral motor adaptation that was complete by day 15. Three days before the end of training, intracortical microstimulation was performed to identify the regions of primary motor cortex responsible for execution of shoulder adduction movements on the experimental (right) and control (left) sides of the brain. A retrograde neuronal tracer was injected into these cortical regions and the animals were returned to the training cage. Following training, small blocks of thalamic tissue containing retrogradely labeled cells were removed from the brains for ultrastructural analyses of presumed cerebellothalamic synapses (see Materials and Methods section). The only ultrastructural change observed to occur in association with the volitional motor adaptation was an increase in the proportion of dendritic shaft active zone with docked synaptic vesicles. PMID- 10363713 TI - Differential localization of Ca2+ channel alpha1 subunits in the enteric nervous system: presence of alpha1B channel-like immunoreactivity in intrinsic primary afferent neurons. AB - Immunocytochemistry was employed to locate calcium (Ca2+) channel proteins in the enteric nervous system (ENS) of the rat and guinea pig. Anti-peptide antibodies that specifically recognize the alpha1 subunits of class A (P/Q-type), B (N type), C and D (L-type) Ca2+ channels were utilized. Alpha1B channel-like immunoreactivity was abundant in both enteric plexuses, the mucosa, and circular and longitudinal muscle layers. Immunoreactivity was predominantly found in cholinergic varicosities, supporting a role for Ca2+ channels, which contain the alpha1B subunit, in acetylcholine release. Immunoreactivity was also associated with the cell soma of calbindin-immunoreactive submucosal and myenteric neurons, cells that have been proposed to be intrinsic primary afferent neurons. Alpha1C channel-like immunoreactivity was distributed diffusely in the cell membrane of a large subset of neuronal cell bodies and processes, whereas alpha1D was found mainly in the cell soma and proximal dendrites ofvasoactive intestinal polypeptide-immunoreactive neurons in the guinea pig gut. Alpha1A channel-like immunoreactivity was found in a small subset of cell bodies and processes in the rat ENS. The differential localization of the alpha1 subunits of Ca2+ channels in the ENS implies that they serve distinct roles in neuronal excitation and signaling within the bowel. The presence of alpha1B channel-like immunoreactivity in putative intrinsic primary afferent neurons suggested that class B Ca2+ channels play a role in enteric sensory neurotransmission; therefore, we determined the effects of the N-type Ca2+ channel blocker, omega-conotoxin GVIA (omega-CTx GVIA), on the reflex-evoked activity of enteric neurons. Demonstrating the phosphorylation of cyclic AMP (cAMP)-responsive element-binding protein (pCREB) identified neurons that became active in response to distension. Distension elicited hexamethonium-resistant pCREB immunoreactivity in calbindin immunoreactive neurons in each plexus; however, in preparations stimulated in the presence of omega-CTx GVIA, pCREB immunoreactivity was found only in calbindin immunoreactive neurons in the submucosal plexus and not in myenteric ganglia. These data confirm that intrinsic primary afferent neurons are located in the submucosal plexus and that N-type Ca2+ channels play a role in sensory neurotransmission. PMID- 10363714 TI - Central vasotocin-immunoreactive system in a male passerine bird (Junco hyemalis). AB - Previous investigations have identified regions of the avian brain that contain immunoreactive vasotocinergic (VT-ir) cell bodies and fibers. These studies exclusively used domesticated species, and the relevance of the findings for free living birds has not been established. The present study used immunocytochemistry to determine the neuroanatomical distribution of the VT-ir system in the brain of a well-studied male passerine bird (dark-eyed junco, Junco hyemalis) obtained from a natural population in interior Alaska (65 degrees N, 147 degrees W). VT-ir cell bodies were observed in several brain regions (paraventricular and supraoptic nuclei, nucleus of the stria terminalis), where they have been described in other oscine species. VT-ir fibers were widespread in many brain regions and were especially abundant in the medial preoptic nucleus, the basal region of the septum, and the hypothalamic-neurohypophyseal tract. Fibers were also present in brain regions that are involved in the control of vocal behavior including the ventromedial capsular region of the nucleus robustus archistriatalis and the dorsomedial portion of the mesencephalic nucleus intercollicularis. The widespread brain distribution of VT-ir cell bodies and fibers in juncos generally resembles that of domestic birds and suggests a role for this neuropeptide in the control of reproductive behavior and physiology. PMID- 10363715 TI - Cellular expression of alpha-gustducin and the A blood group antigen in rat fungiform taste buds cross-reinnervated by the IXth nerve. AB - Although taste buds are trophically dependent on their innervation, cross reinnervation experiments have shown that their gustatory sensitivities are determined by the local epithelium. Both the gustatory G-protein, alpha gustducin, and the cell-surface carbohydrate, the A blood group antigen, are expressed by significantly fewer fungiform than vallate taste cells in the rat. In these experiments, one side of the anterior portion of the tongue was cross reinnervated by the IXth nerve in order to determine whether the molecular expression of taste bud cells is determined by the epithelium from which they arise or by the nerve on which they are trophically dependent. The proximal portion of the IXth nerve was anastomosed to the distal portion of the chorda tympani (CT) nerve using fibrin glue (IX-CT rats). Control animals had the CT cut and reanastomosed using the same technique (CT-CT rats), or had the CT avulsed from the bulla and resected to prevent regeneration (CTX rats). The animals survived for 12 weeks postoperatively, and the tongues were removed, stained with methylene blue, and the fungiform taste pores counted on both sides. Tissue from the anterior 5 mm of the tongue was cut into 50-microm sections, which were incubated with antibodies against alpha-gustducin and the human blood group A antigen. In both CT-CT and IX-CT rats, there was regeneration of fungiform taste buds, although in both groups there were significantly fewer taste buds on the operated side of the tongue. The normal vallate papilla had a mean of 8.37 alpha gustducin-expressing cells and 5.22 A-expressing cells per taste bud, whereas the fungiform papillae contained 3.06 and 0.23 cells per taste bud, respectively. In both CT-CT and IX-CT rats there was a normal number of cells expressing alpha gustducin or the A antigen in regenerated taste buds; in the CTX animals there was a significant decrease in the expression of these markers. These results demonstrate that the molecular phenotype of taste bud cells is determined by the local epithelium from which they arise and not by properties of the innervating nerve. PMID- 10363716 TI - Origin of thalamic inputs to the primary, premotor, and supplementary motor cortical areas and to area 46 in macaque monkeys: a multiple retrograde tracing study. AB - The origin of thalamic inputs to distinct motor cortical areas was established in five monkeys to determine whether the motor areas receive inputs from a common thalamic nucleus and the extent to which the territories of origin overlap. To not rely on the rough definition of cytoarchitectonic boundaries in the thalamus, monkeys were subjected to multiple injections of tracers (four to seven) in the primary (M1), premotor (PM), and supplementary (SMA) motor cortical areas and in area 46. The cortical areas were distributed into five groups, each receiving inputs from a specific set of thalamic nuclei: 1) M1; 2) SMA-proper and the caudal part of the dorsal PM (PMdc); 3) the rostral and caudal parts of the ventral PM (PMvr and PMvc); 4) the rostral part of the dorsal PM (PMdr); and 5) the superior and inferior parts of area 46 (area 46sup and area 46inf). A major degree of overlap was obtained for the origins of the thalamocortical projections directed to areas 46inf and 46sup and for those terminating in SMA-proper and PMdc. PMvc and PMvr received inputs from adjacent and/or common thalamic regions. In contrast, the degree of overlap between M1 and SMA was smaller. The projection to M1 shared relatively limited zones of origin with the projections directed to PM. Thalamic inputs to the motor cortical areas (M1, SMA, PMd, and PMv), in general, were segregated from those directed to area 46, except in the mediodorsal nucleus, in which there was clear overlap of the territories sending projections to area 46, SMA-proper, and PMdc. PMID- 10363717 TI - Organisation of reciprocal connections between trigeminal and vestibular nuclei in the rat. AB - In order to study the connection patterns between the sensory trigeminal and the vestibular nuclei (VN), injections of anterogradely and/or retrogradely transported neuronal tracers were made in the rat. Trigeminal injections resulted in anterogradely labelled fibres, with an ipsilateral preponderance, within the VN: in the ventrolateral part of the inferior nucleus (IVN), in the lateral part of the medial nucleus (MVN), in the lateral nucleus (LVN) with a higher density in its ventral half, and in the superior nucleus (SVN), more in the periphery than in the central part. Moderate trigeminal projections were observed in the small vestibular groups f, x and y/l and in the nucleus prepositus hypoglossi. Additional retrogradely labelled neurones were seen in the IVN, MVN, and LVN, in the same regions as those receiving trigeminal afferents. Morphological analysis of vestibular neurones demonstrated that vestibulo-trigeminal neurones are relatively small and belong to a different population than those receiving projections from the trigeminal nuclei. The trigeminovestibular and vestibulo trigeminal relationships were confirmed by tracer injections in the VN. The results show that, in the VN, there is sensory information from facial receptors in addition to those reported from the neck and body. These facial afferents complement those from the neck and lower spinal levels in supplying important somatosensory information from the face and eye muscles. The oculomotor connections of the respective zones of the VN receiving trigeminal afferents suggest that sensory inputs from the face, including extraocular proprioception, may, through this pathway, influence the vestibular control of eye and head movements. PMID- 10363718 TI - Definition of the alpha 2 region of HLA-DR molecules involved in CD4 binding. AB - HLA class II molecules present antigenic peptides to the T cell receptor of CD4+ T lymphocytes and interact with CD4 during the antigen recognition process. A major CD4 binding site encompassing amino acids (aa) 134-148 in the beta 2 domain of HLA-DR has been previously identified and residues located within the alpha 2 subunit of murine MHC class II I-Ad molecules have been shown to contribute to CD4-class II interaction. To characterize the alpha 2 region of HLA-DR molecules involved in the binding of CD4, we have synthesized overlapping linear and cyclic peptides derived from a region encompassing aa 121-143. We demonstrate that two linear peptides (aa 124-138 and 130-143) and a cyclic one (aa 121-138) specifically bind to CD4-sepharose affinity columns. Although cyclic analogues exhibit more ordered populations as detected by circular dichroism measurements, cyclization did not improve the activity of some peptides. Peptide sequence positioning in HLA-DR1 dimer model indicates that alpha 2 residues 124 to 136 form a solvent-exposed loop which faces the beta 2 loop delimited by residues 134 148. These data suggest that one CD4 molecule contacts both alpha 2 and beta 2 loops of the HLA-DR homodimer. PMID- 10363719 TI - The antigen binding domain of non-idiotypic human anti-F(ab')2 autoantibodies: study of their interaction with IgG hinge region epitopes. AB - In previous studies we described a natural human IgG-anti-F(ab')2 autoantibody family with immunoregulatory properties. Genes coding for the variable regions of the heavy and light chains of the Abs were isolated from a natural Ig gene library and scFv Abs were expressed in E. coli. The scFv Abs bound to F(ab')2 but not to Fab fragments. This points to an epitope located in the hinge region since Fab fragments are lacking most of the hinge. In order to verify our hypothesis, double chain peptides comprising the lower-, middle-, and part of the upper hinge subregion of IgG1-IgG4 were synthesized on cellulose membranes and tested for binding to the Abs. The results show binding of Abs to IgG1 and IgG4 hinge region peptides. In order to identify the key residues of the discontinuous epitopes we carried out complete substitutional analyses in which each amino acid of the wt peptides was substituted by all other amino acids except cysteine. The exchange of proline in the IgG1 or IgG4 middle hinge region abrogated the binding, revealing the importance of this subregion for epitope expression. No binding to the IgG2 or IgG3 hinge was detected. These results indicate that scFv anti F(ab')2 Abs recognize the hinge region of IgG1 and IgG4 and that the expression of the epitope depends on an intact middle hinge subregion. PMID- 10363720 TI - TCR repertoire of suppressor CD8+CD28- T cell populations. AB - The cellular basis of graft rejection and the development of strategies for specific suppression of T cell responses against allogeneic and xenogeneic transplants represents an area of active investigation. Recently, a population of MHC-class I restricted CD8+CD28- T suppressor cells (Ts) which are able to inhibit specifically the proliferative response of allospecific, xenospecific and nominal-antigen specific CD4+ T helper cells (Th) has been identified. We have studied the TCR V beta gene repertoire expressed by CD8+CD28- Ts isolated from allospecific, xenospecific, and nominal antigen-specific T cell lines (TCL). A limited V beta repertoire has been found in all TCLs studied. The most restricted TCR V beta usage was observed within the population of Ts from xenospecific TCLs. The TCR V beta usage within the Ts subset of TCL differs from the TCR repertoire expressed by the CD4+ Th subset of the same TCL. This is consistent with the fact that Ts and Th cells recognize distinct MHC/ antigen complexes. The finding that the TCR repertoire used by Ts is limited opens new avenues for studying the mechanisms of transplant rejection. PMID- 10363721 TI - Nitric oxide and IL-10 production induced by PIII--a fraction of Schistosoma mansoni adult worm antigenic preparation--associated with downregulation of in vitro granuloma formation. AB - Identification and characterization of Schistosoma mansoni antigens that can provide protective immunity, as well as an investigation of their role in host parasite interaction, is crucial for understanding the complex immunoregulatory events that modulate granuloma formation in schistosomiasis. Previous work by our laboratory identified a fraction of S. mansoni soluble adult worm antigenic preparation (SWAP), named PIII, obtained by anionic chromatography on fast protein liquid chromatography (FPLC). This fraction was able to elicit significant in vitro cell proliferation and at the same time lower in vitro and in vivo granuloma formation when compared either to SWAP or to soluble egg antigens (SEA). In the present work, we investigate some biological activities of PIII, such as the stimulation of nitric oxide (NO) and cytokine production. Our data demonstrated that SEA, SWAP and specially PIII were able to induce a time dependent increased NO production during in vitro granuloma reaction. Besides that, PIII evoked increased IL-10 production, but not IL-2 or IFNgamma. Collectively, our results indicate the possibility that the modulation role of PIII on in vitro granuloma might be mediated in part by its ability to induce the higher production, initially of IL-10, and lately of NO. PMID- 10363722 TI - Ligation of MHC class II molecules differentially upregulates TNF beta gene expression in B cell lines of different MHC class II haplotypes. AB - Although the production of selected cytokines by B cells is important for their regulation, little is known about MHC class II-induced cytokine expression in these cells. We designed the present studies to investigate MHC class II-mediated TNF-beta gene expression in 19 EBV-transformed homozygote B cell lines at similar stage of differentiation but presenting different MHC class II haplotypes. Our results demonstrate that in contrast to PMA, engagement of MHC class II with staphylococcal enterotoxin A (SEA), a natural ligand, or with anti-HLA-DR mAb L243, stimulates TNF-beta gene expression in some but not all B cell lines. The differential stimulation of TNF-beta gene expression via MHC class II was not due to the cells MHC class II expression level, nor to their capacity to bind the ligands as evidenced by SEA binding affinity studies. Together these results demonstrate that ligation of MHC class II molecules can stimulate TNF-beta gene expression in a B cell line-dependent manner. The differential cytokine gene expression might be due to an influence of MHC class II haplotype either by a linkage disequilibrium with TNF-beta gene or by a differential association with effector or cell surface molecules. PMID- 10363723 TI - Differential surface expression of MICA by endothelial cells, fibroblasts, keratinocytes, and monocytes. AB - MICA is a new, highly divergent and polymorphic HLA-related gene that has a similar intron-exon organization as the HLA class I genes. It functions as a restriction element for intestinal gammadeltaT cells and it behaves as a cell stress molecule. It is likely that the polymorphic MICA molecule may be target for specific antibodies and T cells in solid organ grafts or in graft versus host disease (GVHD). Previously, we generated three MICA-specific sera in rabbits, which were used for Western blot, flow cytometry and immunoprecipitation. We demonstrated that MICA is expressed in endothelial cells, keratinocyes and monocytes, but not in CD4+, CD8+ or CD19+ lymphocytes. We also found that MICA is expressed on the cell surface in HeLa cells. In the present work, performing peptide neutralization assays, we further confirmed the specificity of the reactivity of these sera against MICA. Also, by Western blot we demonstrate that freshly isolated human skin-derived fibroblasts express MICA. We also investigated the surface expression of MICA in different, freshly-isolated cells. The results show that endothelial cells and fibroblasts express MICA at the cell surface. Although expressing the 62 kDa MICA band, as detected by Western blots, keratinocytes and monocytes do not seem to express this antigen on the cell membrane. This differential surface expression of MICA by endothelial cells and fibroblasts vs. keratinocytes and monocytes, may indicate that the levels of surface MICA are differentially regulated in different cells. Moreover, the expression of MICA on the surface of endothelial cells makes this polymorphic molecule a possible target during the immune response of graft rejection in organ transplantation. PMID- 10363724 TI - Analysis of the cytokine production by cord and adult blood. AB - To date, over 400 human umbilical cord blood cord blood (CB) transplants have been reported from different centres world-wide and it is generally agreed that CB represents an encouraging alternative to bone marrow (BM) transplantation. There are a variety of reasons for this which include the wider availability and easier access of CB compared to BM. In addition it has been suggested that there is a reduced graft-versus-host-disease (GvHD) with CB compared to BM transplantation. The explanations for this implied benefit are numerous, but research into this area is only just beginning. Nevertheless, it is clear that both T cells and natural killer (NK) cells have reduced function when isolated from CB compared to adult and both these cell types have been implicated in GvHD pathogenesis. How and why the function is reduced is yet to be determined. Many laboratories have tried to answer these questions and the majority have done this by comparing the function of lymphocytes obtained from adult blood with those compared with CB. Since cytokine production by a cell is an indication of the cells function it is important to determine the differences between adult and CB with respect to production of these soluble factors. Here, we have reviewed the current research regarding these CB and adult cell comparisons with an emphasise on cytokine production. Our aim is to obtain a clearer understanding of the mechanisms which may be involved in causing a reduced GvHD in CB compared to BM transplantation. PMID- 10363725 TI - HLA and susceptibility to cervical neoplasia. AB - The association between cervical neoplasia and certain HLA phenotypes observed in different studies has not been consistent. By serological typing, the association between HLA antigens, cervical carcinoma and cervical intraepithelial neoplasia (CIN) was studied in a group of 172 and 116 patients, respectively. We demonstrated an increased frequency of B63 in patients with HPV types other than HPV 16 or 18, and B55 in patients that were negative for all HPV types. The association between cervical carcinoma and DQ3, described in various populations, was not observed in the present study. However, we confirmed other previously observed associations between cervical cancer and class II antigens, i.e., a positive correlation with DR15 irrespective of the HPV status, with DR3 in patients harboring HPV types other than HPV 16 or 18, and with DR11 among HPV 16 positive patients. In contrast, a negative correlation between DR13 and HPV positive cervical cancer was observed which suggests protection of this antigen against HPV-associated cervical cancer. A slight increase of DR15 and DQ4 antigens was observed in CIN patients, suggesting that these specific HLA antigens may be important in determining the risk of CIN. PMID- 10363726 TI - CA repeat allele polymorphism in the first intron of the human interferon-gamma gene is associated with lung allograft fibrosis. AB - Interferon-gamma (IFN-gamma) is an inflammatory cytokine that has been implicated in the development of fibrosis in inflamed tissues. In this study we have analysed the association between genetically-determined high IFN-gamma production and development of fibrosis in lung transplants. The human IFN-gamma gene has a variable length CA repeat in the first intron. Our previous study showed that polymorphism of this microsatellite is associated with individual variation in the levels of IFN-gamma production. In vitro production of IFN-gamma showed significant correlation with presence of allele #2 (p < 0.01). In this study allele #2 was found to be associated with allograft fibrosis defined by transbronchial biopsy. An analysis of two groups of lung transplant recipients showed a significant increase in the frequency of allele #2 in the group which developed fibrosis after transplantation compared to the group that did not (p < 0.005). We postulate that the production of IFN-gamma, which is under genetic control, can influence the development of fibrosis in lung allografts. PMID- 10363728 TI - Nomenclature for factors of the HLA System, 1998. PMID- 10363727 TI - A special report: histocompatibility testing guidelines for hematopoietic stem cell transplantation using volunteer donors. Quality Assurance and Donor Registries Working Groups of the World Marrow Donor Association. AB - The World Marrow Donor Association has formulated guidelines for establishing the extent and quality of histocompatibility testing for unrelated donor registries, umbilical cord blood banks, and transplant centers involved in international exchange of hematopoietic stem cells for allogeneic transplantation. Registry and cord blood bank guidelines suggest that, at a minimum, initial HLA typing should be performed for three HLA loci, HLA-A, -B, and -DR, at low resolution/split antigen level. DNA-based testing methods should be utilized for HLA-DR typing. DNA-based testing for HLA-A and -B should replace serologic testing of new volunteer donors and cord blood units as robust protocols and reagents become available to the laboratories. Transplant center guidelines for typing of patient, family and to confirm the HLA types of potential unrelated donors should include, at the minimum, typing HLA-A, B, and -DR loci using primarily DNA-based testing methods at allele level resolution for DRB1 and low resolution/split antigen level for HLA-A and -B. It is strongly recommended that the typing of a patient and the selected donor be performed using the same set of reagents, methodology, and interpretation criteria with fresh tissue samples to ensure HLA identity. Guidelines for laboratory accreditation, approaches to quality assurance and quality control for HLA testing, and suggestions for the format of the HLA database of donor types are also outlined. PMID- 10363729 TI - The role of fluoxetine in the treatment of premenstrual dysphoric disorder. AB - Many women experience psychological and physical symptoms associated with the menstrual cycle, commonly referred to as premenstrual syndrome (PMS). For the 3% to 5% of women who meet Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria for premenstrual dysphoric disorder (PMDD), symptoms are severe and impair social and occupational functioning. Although the etiology of PMDD is unknown, symptoms of dysphoria, including depression and anxiety, predominate and indicate a link to serotonergic neurotransmission. Pharmacotherapy trials have shown greater efficacy with serotonergic versus nonserotonergic compounds. We reviewed the published literature and found 7 controlled and 4 open-label clinical trials of fluoxetine, a selective serotonin reuptake inhibitor, in the treatment of PMDD. These trials demonstrate that PMDD symptoms decreased during treatment with fluoxetine. Preliminary findings suggest that intermittent luteal-phase fluoxetine dosing may also be a suitable treatment strategy for selected patients with PMDD. At 20 mg/d, adverse events were usually transient, rarely caused discontinuation, and were consistent with fluoxetine's known safety profile. Fluoxetine 20 mg/d is an effective and well-tolerated treatment for women with PMDD, a severe variant of PMS. PMID- 10363730 TI - An extended-release formulation of oxybutynin chloride for the treatment of overactive urinary bladder. AB - Detrusor instability, or urinary incontinence, is common in elderly patients, particularly elderly women. The clinical symptoms of overactive, or unstable, urinary bladder include urge urinary incontinence, urgency, and frequency. Mixed urinary incontinence, which comprises urge urinary incontinence and stress incontinence, is manifested by increased intraabdominal pressure on coughing or sneezing. The detrusor muscle of the bladder is under the control of the parasympathetic, or muscarinic, nervous system. The drug of choice in this condition is oxybutynin chloride, which has the ability to block acetylcholine released from parasympathetic nerves in the urinary bladder, preventing contractions of the muscle and exerting a direct spasmolytic effect on the bladder. A new extended-release oral tablet formulation, OROS oxybutynin, uses osmotic pressure to deliver the drug at a controlled rate over approximately 24 hours. It resembles a conventional tablet but has a two-part core consisting of a drug layer and below it, a "push" layer containing osmotically active components, the whole surrounded by a semipermeable membrane with a laser-drilled opening in the drug side. Water in the gastrointestinal tract enters the tablet and mixes with the drug to form a suspension. The "push" layer expands and pushes the suspended drug out of the orifice and into the gastrointestinal tract for eventual absorption. Pharmacokinetic studies have indicated a slow rise in mean plasma concentration of the isomer R-oxybutynin for 4 to 6 hours after a single dose of OROS oxybutynin, followed by maintenance of steady concentrations for up to 24 hours, minimizing the fluctuations between peak and trough associated with TID dosing of 5-mg immediate-release oxybutynin tablets. Efficacy and safety studies comparing the extended-release with the immediate-release formulation of oxybutynin demonstrated equivalent efficacy in patients with overactive urinary bladder. The adverse-event profile of oxybutynin is similar to that of a typical anticholinergic agent such as atropine--dry mouth, constipation, somnolence, blurred vision, headache, and gastrointestinal pain--although in 2 clinical studies, the incidence of dry mouth was less with the extended-release formulation. Once-daily dosing with OROS oxybutynin appears to be well tolerated and effective, as well as convenient, for the treatment of overactive bladder, particularly for elderly patients using multiple medications. PMID- 10363731 TI - A placebo-controlled comparison of the antidepressant efficacy and effects on sexual functioning of sustained-release bupropion and sertraline. AB - Sexual dysfunction, a frequently reported side effect of many antidepressants, may result in patient dissatisfaction and noncompliance with treatment regimens. This paper describes the results of the first placebo-controlled comparison of the efficacy, safety, and effects on sexual functioning of sustained-release bupropion (bupropion SR) and the selective serotonin reuptake inhibitor sertraline. This randomized, double-masked, double-dummy, parallel-group, multicenter trial enrolled 360 patients with moderate-to-severe recurrent major depression. Patients were treated with bupropion SR 150 to 400 mg/d, sertraline 50 to 200 mg/d, or placebo for up to 8 weeks. Patients' depression and sexual functioning were assessed at weekly or biweekly clinic visits; safety was assessed by regular monitoring of adverse events, vital signs, and body weight. Treatment groups were similar at baseline in terms of age, sex, and race, and most patients had a diagnosis of moderate uncomplicated depression. Patients treated with bupropion SR or sertraline showed similar improvements on all efficacy measures; both active treatments were superior to placebo in improving scores on all rating scales for depression at various time points. Significantly more patients treated with sertraline experienced orgasmic dysfunction throughout the study than did patients treated with bupropion SR or placebo (P < 0.001). Headache was the most frequently reported adverse event in all 3 treatment groups and occurred with similar frequency in each group (30% to 40%). Nausea (31%), diarrhea (26%), insomnia (18%), and somnolence (17%) occurred in significantly more patients in the sertraline group than in the bupropion SR group (18%, 7%, 13%, and 3%, respectively) and the placebo group (10%, 11%, 4%, and 6%, respectively). Dry mouth occurred more frequently with bupropion SR (19%) than with sertraline (14%) or placebo (12%), although the differences were not significant. Changes in vital signs were similar in all groups. Similar (small, but not statistically significant) decreases in mean body weight were seen in both the bupropion SR (-1.06 kg) and sertraline (-0.79 kg) groups, whereas the placebo group experienced a minor increase (0.21 kg). Although bupropion SR and sertraline were similarly well tolerated and effective in the treatment of depression, sertraline treatment was more often associated with sexual dysfunction and certain other adverse events compared with bupropion SR and placebo. Therefore, bupropion SR may be an appropriate choice as an antidepressant for the treatment of sexually active patients. PMID- 10363732 TI - Comparison of the upper gastrointestinal safety of Arthrotec 75 and nabumetone in osteoarthritis patients at high risk for developing nonsteroidal anti inflammatory drug-induced gastrointestinal ulcers. AB - A 6-week, multicenter, double-masked, placebo-controlled, parallel-group study compared the upper gastrointestinal (UGI) safety of Arthrotec 75 (diclofenac sodium 75 mg-misoprostol 200 microg; G.D. Searle & Co., Skokie, Illinois) administered twice daily with that of nabumetone 1500 mg administered once daily in 1203 patients with symptomatic osteoarthritis (OA) of the hip or knee. All patients had a documented clinical history of endoscopically confirmed gastric, pyloric-channel, or duodenal ulcer or > or = 10 erosions in the stomach or duodenum. UGI endoscopy was performed at baseline and again at week 6 or early withdrawal. Treatment with Arthrotec 75 resulted in a significantly lower combined incidence of endoscopically confirmed gastric and duodenal ulcers compared with nabumetone (4% vs 11%), and its rate of endoscopically confirmed ulceration was equivalent to that of placebo. The incidence of gastric ulcers alone was also significantly lower with Arthrotec 75 than with nabumetone (1% vs 9%). The incidence of duodenal ulcer with Arthrotec 75 was not significantly different from that with nabumetone (4% vs 3%). Types of adverse events were similar for all treatment groups, with GI adverse events predominating. Arthrotec 75 was well tolerated by the majority of patients. The results of this study demonstrate that Arthrotec 75 has a superior UGI safety profile, causing significantly fewer UGI ulcers, in comparison with nabumetone in patients with symptomatic OA and a documented history of ulcers or > or = 10 erosions. PMID- 10363733 TI - Sparfloxacin versus ciprofloxacin for the treatment of community-acquired, complicated skin and skin-structure infections. AB - Fluoroquinolones have been shown to be effective in the treatment of complicated skin and skin-structure infections, in part because of their broad-spectrum antibacterial activity against causative pathogens that are resistant to older antimicrobial agents. We enrolled 603 adult patients (>58% male, >85% white) in a double-masked, double-dummy, randomized, multicenter trial to compare the efficacy and tolerability of sparfloxacin (400-mg loading dose followed by 200 mg once daily) with those of ciprofloxacin (750 mg twice daily) for 10 days in the treatment of community-acquired, complicated skin and skin-structure infections. The primary efficacy variable was clinical response, based on assessment of signs and symptoms, in the clinically assessable population. Patients in the sparfloxacin and ciprofloxacin groups were comparable with respect to demographic characteristics, underlying diseases, medical history, and laboratory test results. Wound infection was the most common diagnosis, and Staphylococcus aureus was the most frequently isolated pathogen. For the 475 clinically assessable patients, the clinical success rate (percentage of patients cured or improved) was 90.1% (210/233) with sparfloxacin and 87.2% (211/242) with ciprofloxacin. In this analysis (95% confidence interval [CI], -2.8 to 8.6) and the intent-to-treat analyses (95% CI, -4.2 to 6.2), results with sparfloxacin were statistically equivalent to those with ciprofloxacin (95% CI, -1 to 15.3). For bacteriologically assessable patients, eradication rates were 87.0% (141/162) with sparfloxacin and 79.9% (123/154) with ciprofloxacin (95% CI, -1 to 15.3). Eradication rates of S. aureus and coagulase-negative staphylococcal infections were 90.2% (101/112) with sparfloxacin and 77.9% (88/113) with ciprofloxacin. For patients with 2 or more pathogens at baseline (mixed infections), bacteriologic success was 87.6% for sparfloxacin and 77.9% for ciprofloxacin. Pseudomonas aeruginosa infections were eradicated or presumed eradicated in 71.4% (10/14) of sparfloxacin-treated patients and 87.5% (7/8) of ciprofloxacin-treated patients. Overall success rates in the bacteriologically assessable patients for sparfloxacin (84.6% [137/162]) and ciprofloxacin (78.6% [121/154]) were statistically equivalent (95% CI, -2.5 to 14.5). Tolerability was assessed in all patients who received study medication. The overall frequency of treatment related adverse events was comparable in the 2 treatment groups (26.5% sparfloxacin, 23.3% ciprofloxacin). Drug-related adverse events involving the digestive system occurred in 7.1% of sparfloxacin-treated patients and 19.0% of ciprofloxacin-treated patients; photosensitivity reactions were reported in 11.1% of patients in the sparfloxacin group and 0.7% of patients in the ciprofloxacin group (P < 0.001). The mean change in QTc interval from baseline to the maximum on-treatment value was greater in the sparfloxacin group (9 milliseconds) than in the ciprofloxacin group (3 milliseconds) (P = 0.005; 95% CI, 0.002 to 0.010). The efficacy of sparfloxacin was comparable to that of ciprofloxacin in the treatment of community-acquired, complicated skin and skin-structure infections, including those caused by staphylococci, the most common pathogens. Sparfloxacin's once daily regimen, high skin-tissue penetration, and improved activity against gram positive cocci make it a therapeutic alternative to ciprofloxacin for patients who are not at risk for photosensitivity reactions or adverse events associated with prolongation of the QTc interval. PMID- 10363734 TI - Rabeprazole: pharmacokinetics in patients with stable, compensated cirrhosis. AB - This single-center, open-label study was undertaken to compare the tolerability and pharmacokinetic profiles of rabeprazole, a new proton-pump inhibitor (PPI), in healthy volunteers and in subjects with chronic cirrhosis. Thirteen healthy men and 10 men with stable, compensated cirrhosis documented by biopsy or liver/spleen scan received a single 20-mg rabeprazole dose. Blood samples were drawn before and up to 24 hours after drug administration for the determination of plasma rabeprazole concentrations using high-performance liquid chromatography. Adverse events, vital signs, electrocardiograms, physical findings, and clinical laboratory test results were monitored before and during treatment to determine how rabeprazole was tolerated. Chronic liver disease substantially altered the pharmacokinetic profile of rabeprazole. The maximum rabeprazole concentration (+/- SD) in subjects with cirrhosis (635+/-199 ng/mL) was approximately 50% higher than that in the healthy volunteers (401+/-246 ng/mL), and both area under the curve and elimination half-life were increased by approximately 100%. Oral clearance in subjects with cirrhosis was 38% of that in the healthy volunteers. Rabeprazole was well tolerated by both groups. Three subjects reported a total of 5 clinical adverse events that were judged as definitely or possibly related to rabeprazole treatment. Some minor changes in laboratory values were judged to be clinically insignificant. In patients with mild-to-moderate liver dysfunction, clearance of this PPI, as with other members of the class, was markedly reduced and plasma levels were increased. Although caution is always warranted in patients with severe liver disease, drug accumulation is unlikely with rabeprazole 20 mg once daily, and dose adjustment does not appear to be indicated in patients with mild-to-moderate liver dysfunction. PMID- 10363735 TI - Repaglinide pharmacokinetics in healthy young adult and elderly subjects. AB - In this open-label, single-center, pharmacokinetic study of repaglinide, 12 healthy volunteers (6 men, 6 women) were enrolled in each of 2 groups (total, 24 volunteers). One group consisted of young adult subjects (18 to 40 years), and the other group consisted of elderly subjects (> or = 65 years). On day 1, after a 10-hour fast, all 24 subjects received a single 2-mg dose of repaglinide. Starting on day 2 and continuing for 7 days, subjects received a 2-mg dose of repaglinide 15 minutes before each of 3 meals. On day 9, subjects received a single 2-mg dose of repaglinide. Pharmacokinetic profiles, including area under the curve, maximum concentration (Cmax), time to Cmax, and half-life, were determined at completion of the single-dose and multiple-dose regimens (days 1 and 9, respectively). Trough repaglinide values were collected on days 2 through 7 to assess steady state. The single-dose and multiple-dose pharmacokinetic variables of serum repaglinide were not significantly different between young adult and elderly subjects. Repaglinide was well tolerated in both groups. Hypoglycemic events occurred in 5 young adult and 5 elderly subjects. This study demonstrates that the pharmacokinetics of repaglinide are similar in healthy young adult and elderly subjects. PMID- 10363736 TI - Loracarbef versus clarithromycin in children with acute otitis media with effusion. AB - Two multicenter, randomized, single-masked, parallel-group studies compared loracarbef and clarithromycin with regard to efficacy, tolerability, and patient acceptance. Three hundred thirty-four children aged 6 months to 3 years with acute otitis media with effusion received loracarbef (15 mg/kg) or clarithromycin (7.5 mg/kg) orally twice daily for 10 days. Patients were assessed for the presence of the diagnostic signs and symptoms of otitis media with effusion by physical examination and pneumatic otoscopy at 48 hours pretreatment, 3 to 5 days after initiation of treatment, 0 to 3 days after the final dose (posttreatment), and 14 to 21 days later (termination). Symptoms were assigned numeric values. Symptomatic response was assessed at the posttherapy and termination visits. Tolerability was determined by assessing adverse events, and a patient acceptance survey was completed by each patient's caregiver. The combined results of these 2 studies showed that the efficacy and tolerability of loracarbef were comparable to those of clarithromycin. Adverse events were reported by 46.4% of loracarbef patients and 41.0% of clarithromycin patients, with no statistically significant difference between groups. In the intent-to-treat analysis, 57.9% of loracarbef patients were cured at the termination of the study, compared with 55.7% of clarithromycin patients. Improvement was seen in 4.1% of loracarbef patients and 2.7% of clarithromycin patients. Results of the patient acceptance survey showed a clear preference for loracarbef over clarithromycin. Difficulties with administration of treatment were reported by 36.3% of clarithromycin caregivers, compared with 7.8% of loracarbef caregivers (P < 0.001). A desire to stop treatment was reported by 23.8% of clarithromycin caregivers, compared with 7.8% of loracarbef caregivers (P < 0.001). Taste and texture issues were most frequently cited as reasons for nonacceptance. PMID- 10363738 TI - Nefazodone therapy in patients with treatment-resistant or treatment-intolerant depression and high psychiatric comorbidity. AB - Given the potentially severe functional impairment, morbidity, and high costs associated with refractory depression, it is important to explore all treatment options that may benefit patients with this disorder. This is a retrospective, uncontrolled analysis of our experience with nefazodone therapy in treatment resistant and treatment-intolerant depression. Potential candidates for nefazodone therapy were referred by their treating psychiatrist. Documentation of failure to respond to previous antidepressant therapy, a diagnosis of clinical depression according to criteria in the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, and completion of a Beck Depression Inventory (BDI) were required before initiation of nefazodone. A follow-up BDI was obtained after > or =4 weeks of nefazodone therapy. A Clinical Global Inventory (CGI) score was obtained retrospectively based on documentation of target symptoms in the clinical record of the last clinic visit. The study group consisted of 20 patients with treatment-resistant or treatment-intolerant major depression who received nefazodone therapy. The mean (+/- SD) age of the group was 48.1+/-9.4 years. The mean number of previously failed antidepressant trials was 1.9+/-0.6. Psychiatric comorbidity in this group was substantial, with posttraumatic stress disorder (PTSD) found in 11 (55%) patients, substance abuse in 3 (15%) patients, and personality disorder found in 2 (10%) patients. After treatment with nefazodone, 11 of 20 patients (55%) were rated on the CGI as much or very much improved. In addition, 9 patients (45%) had >20% improvement on BDI, 3 patients (15%) had 10% to 20% improvement, and 6 patients (30%) had <10% change. Two patients (10%) discontinued nefazodone therapy due to adverse effects. Analysis of our experience with nefazodone therapy in a population with treatment resistant depression and a high degree of psychiatric comorbidity suggests that approximately 50% of patients may have substantial response to treatment, with a smaller proportion having a more modest clinical response. While receiving nefazodone therapy, most patients continued to take concurrently prescribed psychotropic medications, primarily anxiolytics or other antidepressants. Of interest was the positive drug response among a subgroup of individuals with depression and chronic, severe PTSD. Larger, controlled studies are needed to determine whether these preliminary observations are confirmed. PMID- 10363737 TI - Efficacy and safety profile of ketorolac 0.5% ophthalmic solution in the prevention of surgically induced miosis during cataract surgery. AB - This multicenter, double-masked, randomized, parallel study compared the efficacy and safety profile of ketorolac tromethamine 0.5% ophthalmic solution with that of its vehicle in the maintenance of pupillary mydriasis during cataract surgery. A total of 176 adult patients scheduled to undergo unilateral extracapsular cataract extraction and posterior-chamber intraocular lens implantation received either ketorolac tromethamine 0.5% (n = 89) or vehicle (n = 87), starting 2 hours before surgery. One drop of study medication was instilled every 30 minutes for a total of 4 drops. No epinephrine was used in the intraoperative irrigating solution. Pupil diameter was measured with a caliper at 3 time points during surgery. To ensure participant safety, biomicroscopy, ophthalmoscopy, intraocular pressure, adverse events, and preoperative and postoperative visual acuity and refractive error were also monitored. The mean change in horizontal and vertical pupil diameter from the time of the first incision to after cortical irrigation and aspiration was significantly less with active ketorolac than with vehicle (P < or = 0.014). Consequently, mean pupil diameter after cortical irrigation and aspiration was significantly greater with ketorolac than with vehicle (P < or = 0.030). No significant between-group differences were observed in the change in pupil diameter between the end of surgery and postoperative administration of a miotic agent, safety variables, or occurrence of adverse events. In this study, ketorolac tromethamine 0.5% ophthalmic solution provided effective and well tolerated inhibition of surgically induced miosis during cataract surgery. PMID- 10363739 TI - Safety and efficacy of atovaquone and proguanil hydrochloride for the prophylaxis of Plasmodium falciparum malaria in South Africa. AB - The objective of this study was to determine the safety and efficacy of atovaquone and proguanil hydrochloride combination therapy for the prophylaxis of Plasmodium falciparum malaria in at-risk nonimmune subjects in South Africa. This open-label trial was conducted at research sites in South Africa during the main malaria transmission season, February through July. The study volunteers were temporarily living in, or traveling to, a malaria-endemic area. They received I tablet of 250 mg atovaquone and 100 mg proguanil hydrochloride once daily for up to 10 weeks. Subjects were monitored using sequential clinical and laboratory assessments. Thick blood smears were stained and evaluated by a central laboratory. An immunochromatographic test for P. falciparum was also used for on site patient management. Prophylactic success was summarized using a 95% confidence interval for the proportion of subjects who did not develop parasitemia or who withdrew due to a treatment-related adverse event. A total of 175 subjects (15% women) were enrolled in the trial. The mean duration of drug exposure was 8.9 weeks. The combination of atovaquone and proguanil hydrochloride was well tolerated. The most frequently reported adverse events considered possibly related to study treatment were headache (7%), abdominal pain (2%), increased cough (2%), and skin disorder (2%). No serious adverse events were reported, and no treatment-emergent effects were noted for any laboratory variables. One subject who was noncompliant with therapy developed parasitemia, and 3 subjects withdrew due to a treatment-related adverse event (2 subjects with headache and 1 with nausea and dizziness). The prophylaxis success rate was 97%. In this study, atovaquone and proguanil hydrochloride combination therapy had an excellent safety and efficacy profile for prophylaxis of P. falciparum malaria in nonimmune subjects. PMID- 10363740 TI - Impact of palivizumab on expected costs of respiratory syncytial virus infection in preterm infants: potential for savings. AB - In its clinical assessment of the respiratory syncytial virus (RSV)-specific monoclonal antibody palivizumab, the IMpact-RSV Study Group demonstrated a reduction in hospitalizations for RSV-related lower respiratory tract infection in infants who received prophylaxis compared with infants who did not receive prophylaxis. An assessment of the RSV-related expenses for managing both groups of infants is needed to provide insight into the value of prophylaxis. The present study was conducted to identify and compare RSV-related health care expenditures incurred by infants who did not receive prophylaxis throughout one RSV season and after. Using a decision-analytic model populated with data from the contemporary medical literature, a pharmacoeconomic study was conducted from the perspective of the payer. Probabilities for RSV-related hospitalizations of infants who did and did not receive prophylaxis were abstracted from several published studies. Components of inpatient and outpatient care were identified through examination of hospital records, reviews of the published literature, and consultation with expert clinicians. Charges related to prophylaxis and medical management of infection were abstracted from hospital billing records and published data. Appropriate charges were applied to decision-tree branches and multiplied by in-line probabilities for outcomes. Products at terminal nodes were summed to establish total expected charges for both groups of infants. Widespread clinical use of prophylactic palivizumab would result in incremental expenses < or =$3459 per infant or cost savings < or =$39,107 per infant. The variability in value of prophylaxis derives from the rate of RSV-related hospitalizations in the community and the total health care expense of managing infected infants. PMID- 10363741 TI - Meeting the information needs of drug purchasers: the evolution of formulary submission guidelines. AB - The emergence of formulary submission guidelines in the 1990s has been seen by many as an attempt to come to grips with the issue of drug management within health systems and to provide, for the first time, a coherent and methodologically rigorous approach to formulary selection. Judging from the available evidence, however, guidelines have fallen far short of their potential. The purpose of this paper is to consider what role guidelines have played in health system management and whether they play a useful role in providing a methodologically sound basis for drug-impact assessment. Two polar cases in guideline development are examined: the Australian guidelines first published in 1992 and now undergoing a second major revision and the guidelines recently published by Blue Cross and Blue Shield of Colorado and Nevada. The former guidelines represent what can be described as the traditional, clinical paradigm of drug-impact assessment; the latter represent what can be called the system impact paradigm. The argument put forward is that the Australian guidelines are essentially an anachronism, offering little to those whose principal concern is with the management of health systems. In their emphasis on a hierarchy of evidence and a clinical trial-focused, cost-effectiveness evaluation perspective, they represent a methodologic dead end in a number of important respects. The systems-based approach, on the other hand, with its emphasis on validation of claims and the need to consider a range of drug-impact and risk-management scenarios, offers an analytic framework that, while possibly less rigorous, is likely to contribute significantly to the management of health care systems. PMID- 10363742 TI - Relationship between serum ferritin and risk factors for ischaemic heart disease in 2235 Danes aged 30-60 years. AB - OBJECTIVES: The aim was to examine the relationships between serum ferritin and risk factors for ischaemic heart disease (physical activity, body mass index, tobacco smoking, alcohol consumption, serum total cholesterol, serum triglycerides, serum high-density lipoprotein (HDL) cholesterol, systolic and diastolic blood pressures). DESIGN: Epidemiological population survey performed at the Copenhagen County Centre for Prevention of Disease in 1982-84. SUBJECTS: The participants were selected at random from the census register and comprised 2235 healthy Caucasian Danes, all non-blood-donors (1044 men and 1191 women), in cohorts of 30, 40, 50 and 60 years of age. The participants gave a detailed medical history and had a clinical examination including blood samples. MAIN OUTCOME MEASURES: In both men and women, all risk factors displayed a significant increase with age. In men aged 40-60 years, significant positive associations were found between serum ferritin and the following risk factors: body mass index, alcohol intake, serum triglycerides, and systolic and diastolic blood pressures. There was a significant negative association between serum ferritin and tobacco smoking. There was no association between serum ferritin and physical activity, serum total cholesterol or serum HDL cholesterol. In women aged 40-60 years, significant positive associations were found between serum ferritin and the following risk factors: body mass index, alcohol intake and serum triglycerides. There was no association between serum ferritin and physical activity, tobacco smoking, serum total cholesterol, serum HDL cholesterol or blood pressure. CONCLUSIONS: Associations were found between serum ferritin and some risk factors for ischaemic heart disease in men and women. The clinical significance of these findings remains to be clarified. One may hypothesize that the 'missing link' between serum ferritin and ischaemic heart disease in men is the relationship between serum ferritin, serum triglycerides and blood pressure. PMID- 10363743 TI - The circulating renin-angiotensin system during treatment with metoprolol or captopril in patients with heart failure due to non-ischaemic dilated cardiomyopathy. AB - OBJECTIVE: To investigate the effects of beta-blocker (metoprolol) or angiotensin converting enzyme inhibitor (captopril) treatment on neurohormonal function in a randomized prospective study on patients with heart failure due to dilated cardiomyopathy. PATIENTS: Fifty-four patients (42 men and 12 women, mean age 50 years) were studied. There were three patients in NYHA (New York Heart Association) functional class I, 32 patients in class II and 19 patients in class III. METHODS: Measurements of plasma renin activity (PRA). plasma angiotensin II (A II) concentration and plasma atrial natriuretic peptide (ANP) concentration were made at rest and also in a subgroup (n = 32) during exercise. The urinary excretion of aldosterone was also determined. Investigations were performed at baseline, and after 3 and 6 months. Therapy was then stopped and the patients were re-investigated 1 month thereafter. RESULTS: The mean level of PRA was normal at baseline, reduced during therapy with metoprolol, and increased during therapy with captopril. The mean plasma concentration of A II was reduced during exercise and there was a trend towards a reduction even at rest in the metoprolol group, but not in the captopril group. The urinary excretion of aldosterone decreased in both groups. The mean plasma concentration of ANP was elevated at baseline and declined during exercise in the metoprolol group. CONCLUSION: In patients with dilated cardiomyopathy and only a partly activated renin angiotensin system, both metoprolol and captopril reduced urinary excretion of aldosterone. Furthermore, metoprolol suppressed the exercise-induced increase in ANP, suggesting a favourable effect on ventricular performance. PMID- 10363744 TI - Improvement in quality of life differs between women and men after coronary artery bypass surgery. AB - OBJECTIVE: To study improvement in quality of life (QoL) after coronary artery bypass grafting (CABG) in relation to gender. BACKGROUND: Women generally report worse QoL after CABG than men. However, women are older and more symptomatic prior to surgery, which should be considered in comparative analyses. METHODS: We studied consecutive patients who underwent CABG between 1988 and 1991 [n = 2121] with a QoL questionnaire containing the Physical Activity Score, the Nottingham Health Profile and the Psychological General Well-being Index prior to, 3 months, 1 year and 2 years after surgery. RESULTS: Females were older than men with more concomitant diseases preoperatively. QoL was improved on all postoperative occasions for both sexes. Improvement in the Physical Activity Score was somewhat, although not significantly, greater in males. Improvement in the Nottingham Health Profile was greater in females. General well-being showed no consistent pattern for improvement. CONCLUSIONS: QoL is significantly improved after CABG in both sexes throughout follow-up. There is a complex association between improvement in various aspects of QoL and gender. PMID- 10363745 TI - Insulin-like growth factor-I lowers fasting and postprandial triglyceride levels without affecting chylomicron clearance in healthy men. AB - OBJECTIVES: To study whether IGF-I treatment alters the postprandial lipid and lipoprotein metabolism. DESIGN: Randomized, crossover study. SETTING: University Hospital, Zurich, Switzerland. SUBJECTS: Seven young healthy male subjects (aged 27+/-4 years, body mass index (BMI) 21.8+/-1.7 kg m(-2)). INTERVENTIONS: Each subject was studied two times at 2-week intervals, treated with saline 0.9% (S) and IGF-I (8 microg kg(-1) h(-1)) by a continuous subcutaneous infusion. 60 h after the start of treatment a vitamin A loading test was performed after an overnight 12-h fast. MAIN OUTCOME MEASURES: Glucose, insulin, total and free IGF I, FFA, triglycerides and retinyl palmitate, total cholesterol, HDL and LDL cholesterol, lipoprotein (a) and apolipoprotein B were measured in serum before and after the fatty meal. RESULTS: Total IGF-I levels rose from 29.0+/-3.3 nmol L(-1) to 113.3+/-9.0 nmol L(-1) (P<0.02) and free IGF-I from 0.24+/-0.05 to 1.08+/-0.28 nmol L(-1) (P<0.02) during IGF-I treatment. IGF-I administration reduced insulin concentrations by 50% (P<0.02), as assessed by the area under the curve. Serum triglyceride levels were significantly lower at baseline and after the fat load during IGF-I treatment (P<0.02), whereas the retinyl palmitate concentrations in chylomicron and nonchylomicron lipoprotein fractions were similar during both treatment periods. CONCLUSIONS: IGF-I treatment reduces the triglyceride levels most probably by decreasing insulin secretion and the production of VLDL particles, and possibly by increasing their turnover. IGF-I treatment has no significant effect on the metabolism of intestine-derived triglyceride-rich lipoproteins after a high fat meal in healthy young men. PMID- 10363746 TI - The survival and well-being of patients treated for Cushing's syndrome. AB - OBJECTIVE: The main aim of the study was to evaluate the survival, well-being and working capacity of patients treated for Cushing's syndrome. DESIGN: The study was carried out by retrospectively analysing patient records from years 1981-94. Follow-up time was extended from the time of diagnosis to the end of 1996. A questionnaire dealing with symptoms prior to and after therapy, and the quality of life estimated on a Visual Analogue Scale (VAS) was sent to all surviving patients. SETTING: The study was performed in a university hospital. MAIN OUTCOME MEASURES: Survival, subjective well-being, working capacity before and after treatment and disappearance of symptoms after treatment of Cushing's syndrome. RESULTS: During the follow-up time 10 patients died out of a total of 74. The overall standardized mortality ratio (SMR) was 168% (95% CI 81-309%). The SMR of patients with pituitary disease was 267% (89-525%), and in patients with adrenal adenoma it was 135% (16-489%). Forty-six per cent of the surviving patients stated that they felt fully recovered from the disease, but the proportion of patients having persisting symptoms after treatment was also noteworthy. The mean VAS score (range 0-100) was 19 (SD 14) before treatment and 82 (SD 18) after treatment (P<0.001). After treatment, 81% of the patients were able to return to work, 11% retired because of disability, 5% retired because of age and 3% were on sick-leave at the time of answering the questionnaire. During the follow-up time, 42% of the patients with pituitary disease suffered a relapse. However, the effect of the relapse on well-being was not significant. CONCLUSIONS: The mortality risk of patients treated for Cushing's disease was not significantly increased compared with that of the general population. Many symptoms persisted even years after therapy. After clinical recovery, working ability was not always regained. PMID- 10363747 TI - Colonic endocrine cells in patients with familial amyloidotic polyneuropathy. AB - OBJECTIVE: To establish whether the endocrine cell number is affected in the colon in Japanese FAP patients. SETTING: Department of Medicine, Umea University Hospital and Department of Internal Medicine and Pathology, University Hospital, Kumamoto, Japan. SUBJECTS: Autopsy colon tissue specimens from 11 FAP patients and nine controls as well as 12 control biopsy specimens were included in the study. MEASUREMENTS: Endocrine cells in the colon were detected by immunohistochemistry and quantified by computerized image analysis. RESULTS: The autopsy material showed a slight autolysis. Neither enteroglucagon nor pancreatic polypeptide positive cells could be detected in the autopsy material, but were present in biopsy material. There was no statistical difference between autopsy and biopsy specimens regarding the number of peptide YY (PYY), somatostatin and serotonin cells. No significant differences were noted in PYY, somatostatin and serotonin immunoreactive cells in FAP patients compared to autopsy controls, though PYY cells tended to be decreased and serotonin and somatostatin cells tended to be increased in FAP patients. CONCLUSION: The difference between the Swedish and Japanese patients in the endocrine cell content points to the possibility of involvement of other factors than the endocrine cell depletion of the colon might be involved in the pathogenesis of gastro-intestinal dysfunction in FAP. The tendency of PYY to decrease in Japanese FAP might contribute to the development of diarrhoea in these patients. PMID- 10363748 TI - Impaired granulocyte formylpeptide-induced superoxide generation in chronically ill, malnourished, elderly patients. AB - OBJECTIVE: Protein-energy malnutrition (PEM), often occurring in patients with chronic disease, is associated with a decreased capacity to combat infections. In this study we assessed polymorphonuclear neutrophil (PMN) superoxide anion (*O2-) formation in elderly PEM patients with various chronic diseases. DESIGN AND SUBJECTS: Nineteen patients (75+/-1 years), with body mass index 17.1+/-0.4, and 19 age-matched healthy controls were included. Fourteen patients and 14 controls were re-examined in a 3 month follow-up. SETTING: A service of internal medicine at a university-affiliated hospital. INTERVENTIONS: Eight patients were prescribed a dietary liquid supplementation during the observation period. MAIN OUTCOME MEASURES: Superoxide production in PMN induced by fMLP (a receptor ligand) and phorbol myristate acetate (PMA), which acts directly on protein kinase C. RESULTS: fMLP-induced superoxide generation in the malnourished patients was 55+/-5% of that of the controls. However, the patients retained their capacity, 108+/-6% of control PMN generation, to respond to PMA. In those who received formula supplementation, fMLP-generated *O2- production levels were 48+/-8 and 73+/-13% (P = 0.12) of those of controls at the start and after 3 months, respectively. Corresponding figures in those who were not prescribed supplementation were 57+/-8 and 64+/-4% (P = 0.55). CONCLUSION: Possibly contributing to reduced host defence, receptor ligand-induced PMN generation of *O2 is significantly lower in chronically ill, elderly patients with PEM than in age-matched healthy controls. PMID- 10363749 TI - Streptokinase antibodies inhibit reperfusion during thrombolytic therapy with streptokinase in acute myocardial infarction. AB - OBJECTIVES: To evaluate the influence of pretreatment IgG against streptokinase on the outcome of streptokinase treatment in acute myocardial infarction. SETTING: Coronary care unit. DESIGN: From 88 patients admitted to the coronary care unit due to chest pain, blood samples were taken for determination of the pre-existing titre of antibodies against streptokinase. The patients were treated and monitored according to standard protocols. Fifty of the patients received thrombolytic therapy with streptokinase due to acute myocardial infarction and were monitored with continuous dynamic vectorcardiography, making possible the continuous analysis of ST- and QRS-vector changes and determination of the event of reperfusion. None of these 50 patients had been given streptokinase therapy previously. RESULTS: According to the vectorcardiographic criteria 21(42%) patients had signs of early (within 2 h) reperfusion after streptokinase therapy. These patients had lower pre-existing antibody titres than patients without signs of reperfusion (mean values 0.20 and 0.45 arbitrary units, P = 0.01). None of the patients with a titre higher than 0.50 arbitrary units (nine patients) had signs of early reperfusion. Of the 41 patients with a titre lower than 0.50 arbitrary units 52.5% had signs of early reperfusion. CONCLUSION: The present investigation indicates that pre-existing streptokinase antibodies play an important role in reperfusion failure during thrombolytic therapy with streptokinase in acute myocardial infarction. Therefore, the determination of streptokinase antibodies may differentiate between those patients who may benefit from streptokinase treatment and those who should be treated with some other regime. PMID- 10363750 TI - Lean mass and physical activity as predictors of bone mineral density in 16-20 year old women. AB - OBJECTIVE: The aim of the study was to quantify the inter-relationship between bone mineral density and physical activity, muscle strength, and body mass composition in a group of healthy 16-20-year-old women. DESIGN: A cross-sectional study. SETTING: Reykjavik area. SUBJECTS: Two-hundred and fifty-four Icelandic Caucasian women aged 16, 18 and 20 years, randomly selected from the registry of Reykjavik. MAIN OUTCOME MEASURES: Bone mineral content (BMC) and density (BMD) in lumbar spine, hip, distal forearm and total skeleton and lean mass and fat mass were measured with dual energy X-ray absorptiometry (DEXA) and compared with grip strength measured with a dynamometer and physical activity as assessed by a questionnaire. RESULTS: The lean mass had the strongest correlation with BMC and BMD, stronger than weight, height and fat mass, both in univariate analysis (r = 0.41-0.77; P<0.001) and in linear regression analysis. The total skeletal BMD was logarithmically higher by hours of exercise per week (P<0.001)). About 30% of variability in total skeletal BMD in this age group can be predicted by lean mass and physical exercise. CONCLUSIONS: Modifiable factors, such as exercise and adequate muscle seem to be significant predictors of the attainment of peak bone mass in women. PMID- 10363751 TI - Panic disorder in chest pain patients referred for cardiological outpatient investigation. AB - OBJECTIVES: The aims of the study were to: (i) determine the prevalence of panic disorder (PD) in patients referred to cardiological outpatient clinics for evaluation of chest pain; (ii) compare psychiatric comorbidity, psychological distress, pain characteristics and suicidal ideation in PD and non-PD patients: (iii) compare the prevalence of coronary risk factors and medical comorbidity in PD and non-PD patients; and (iv) describe current PD treatment and need for PD treatment as expressed by PD patients. DESIGN: A cross-sectional study based on psychiatric and cardiological investigation. SETTING: Four cardiological outpatient clinics in Oslo, Norway. SUBJECTS: One-hundred and ninety-nine consecutive patients without known heart disease referred to out-patient clinics for investigation of chest pain. MAIN OUTCOME MEASURES: Psychiatric state diagnosis (axis I); scores on self-assessment rating scales of psychological factors and pain modalities; cardiological diagnosis. RESULTS: Thirty-eight per cent of the patients met criteria for current panic disorder (PD). Panic disorder was associated with psychological distress, comorbid psychiatric disorders, medical disorders and significantly higher prevalence of coronary risk factors (P<0.05). Furthermore. the results suggest that these patients were not identified and appropriately treated for panic disorder prior to cardiological investigation. The results indicate that the patients are positive to screening for psychiatric disorder and communicate a need for treatment early in the investigation process. CONCLUSION: PD commonly occurs in this chest pain population. Thus, there is a need to educate physicians caring for these patients about PD identification and treatment. PMID- 10363752 TI - Relationships between bone mineral density, serum vitamin D metabolites and calcium:phosphorus intake in healthy perimenopausal women. AB - OBJECTIVES: To determine the relationships between serum vitamin D metabolites, bone mass, and dietary calcium and phosphorus in a cohort of 510 healthy Danish perimenopausal women. DESIGN: A population-based cross-sectional study. SUBJECTS: A total of 510 healthy women aged 45-58 years, with amenorrhoea for 3-24 months. None of the women was using hormone replacement therapy. MEASUREMENTS: Measurements of total bone mineral content and regional bone mineral density were performed by dual-energy X-ray absorptiometry. Analyses of serum levels of 25-OHD and 1,25-(OH)2D, intact PTH, ionized calcium and phosphate, as well as biochemical markers of bone turnover in blood and urine. Assessment of calcium and phosphorus intake using dietary records. RESULTS: A consistent inverse relationship between serum 1,25-(OH)2D and bone mineral content/ density was found in whole-body mineral content (P = 0.001), spine (P = 0.005) and femoral neck (P<0.05). There was a positive relationship between levels of 1,25-(OH)2D and biochemical bone markers, indicating that high levels of 1,25-(OH)2D are accompanied by increased bone turnover. The dietary calcium:phosphorus ratio was inversely related to serum 1,25-(OH)2D (P = 0.04) and positively related to bone mineral density (P<0.0005). No relationships could be detected between levels of PTH, serum ionized calcium and phosphate, and serum vitamin D metabolites. CONCLUSION: Within normal physiological range, elevated levels of 1,25-(OH)2D were associated with decreased bone mineral density and content, reduced calcium:phosphorus ratio in the diet and increased bone turnover. PMID- 10363753 TI - The spectrum of hepatobiliary disease in primary hypogammaglobulinaemia. AB - OBJECTIVE: To study the prevalence of hepatobiliary disease in a clinically and immunologically well-characterized group of 88 adult Norwegian patients with primary hypogammaglobulinaemia. SUBJECTS: Eighty-eight patients with primary hypogammaglobulinaemia were followed and signs and symptoms of liver disease were recorded. The patients were examined clinically and radiologically on a regular basis with liver biopsies performed when indicated. All patients were tested for hepatitis C virus (HCV) RNA, hepatitis G virus (HGV) RNA and hepatitis B virus (HBsAg). RESULTS: Twenty-one patients were HCV RNA-positive, all having signs of chronic liver disease. Only four patients were HGV RNA-positive, of whom two were also HCV RNA-positive. Amongst the 67 HCV RNA-negative patients, 26 had signs of chronic liver disease, including two who were HGV RNA-positive. HCV RNA-negative patients with liver disease had received intravenous immune globulin substitution more frequently, had a longer history of any form of immune globulin substitution and had a greater incidence of common variable immunodeficiency than patients without signs of liver disease. In most cases (21 of 26 patients) the liver disease was relatively mild. Three patients had granulomatous liver disease, with a relatively aggressive course in all three. CONCLUSION: Hepatobiliary disease is a frequent complication in primary hypogammaglobulinaemia. Liver disease in HCV RNA-negative patients usually has a mild course. HGV does not seem to be a major cause of chronic liver disease in these patients. PMID- 10363754 TI - Evidence of gastrointestinal immune reactivity in patients with sarcoidosis. AB - OBJECTIVES: The aim of the present study was to explore the frequency of clinical and serological manifestations of gastrointestinal immune reactivity in a large group of Swedish patients with sarcoidosis. DESIGN: In patients with documented sarcoidosis, the presence of pernicious anaemia and coeliac disease was examined. Antibodies to H+/K+ ATPase, gliadin (AGA-IgA/IgG) and endomysium (IgA-EMA) were analysed. In H+/K+ ATPase antibody-positive patients, serum gastrin levels were measured and, when elevated, gastrointestinal biopsy was offered (biopsy performed in 6/9 patients): biopsy was also offered to those with positive EMA or AGA of either class (biopsy performed in 8/12 patients). Subjects from national and local studies were used as controls. SETTING: The patients were recruited at the Department of Pulmonary Medicine, and the study was conducted at the Department of Endocrinology, University of Lund, Malmo University Hospital, Malmo, Sweden. SUBJECTS: Of all patients (n = 89) with documented sarcoidosis attending the Department of Pulmonary Medicine between January 1980 and December 1991, 78 [34 females and 44 males; median age at the time of the study, 48 (range 22-81) years; median observation time since the diagnosis of sarcoidosis, 120 (range 1-468) months] were examined. RESULTS: Twenty-nine patients (37.2%) had signs of gastrointestinal immune reactivity. H+/K+ ATPase antibodies were detected in 19 patients (24.4 vs. 4% in controls, P = 0.00015). Serum gastrin levels (median 45, range 22-720 pmol L(-1)) in those patients correlated with antibody titre (r2 = 0.882). Gliadin antibodies were detected in 12 patients (15.4 vs. 8.1% in controls, P = 0.042), of whom 11 (14.1 vs. 4.5% in controls, P = 0.00114) had AGA-IgA alone. One patient had pernicious anaemia and another coeliac disease (EMA-positive). CONCLUSION: We have demonstrated a high frequency of gastric autoimmunity and gluten-associated immune reactivity in patients with sarcoidosis, occurring in almost 40% of the cases, the former being the most frequent gastrointestinal immune manifestation. Despite a high frequency of humoral autoimmunity, the frequencies of clinical disease, pernicious anaemia and coeliac disease were not increased as compared with the control population. PMID- 10363755 TI - Plasma leptin levels are associated with abnormal fibrinolysis in men and postmenopausal women. AB - BACKGROUND: Leptin is a crucial mediator of satiety signals and energy balance, and its circulating levels are increased in obesity. It has recently been shown that plasma leptin levels in humans correlate with circulating insulin and to insulin secretion. This indicates that leptin may be an important link in metabolic consequences of the insulin resistance syndrome. Whether this includes abnormalities in fibrinolysis has not been studied. METHODS AND RESULTS: Healthy subjects (n = 165; 85 men and 80 women) from the Northern Sweden MONICA population were investigated. Anthropometric measurements, oral glucose tolerance tests and sampling for plasma leptin, lipids, fibrinogen and fibrinolytic variables were made. Leptin levels were 342% higher in women than in men and were in both sexes strongly correlated to body mass index (BMI). After adjustments for age and BMI, leptin levels correlated significantly to pre/post glucoseload insulin levels in both sexes. After further adjustment for baseline insulin levels, leptin levels were in males significantly associated with increased waist circumference (P<0.001), low HDL cholesterol (P<0.05), low tPA activity (P<0.01) and high PAI-1 activity (P<0.001). In postmenopausal females, a significant association between leptin and low tPA activity/high PAI-1 activity was seen after adjustment for age and BMI (P<0.05). Conclusions. Circulating levels of leptin are associated with components of the insulin resistance syndrome, including defective fibrinolysis, in men and postmenopausal women. This suggests that leptin may be involved in the mediation of consequences of insulin resistance. PMID- 10363756 TI - Significance of graft occlusion and coronary atherosclerosis 5 years after coronary artery bypass grafting. A quantitative angiographic study with serial exercise testing. AB - OBJECTIVE: To evaluate the relative importance of graft occlusions and progression of atherosclerosis in coronary arteries as causes of the occurrence of angina pectoris and impairment of physical performance 5 years after coronary artery bypass surgery. DESIGN: A 5-year follow-up study. SETTING: University hospital in south-western Finland. SUBJECTS: Altogether, 174 consecutive electively operated bypass patients. MAIN OUTCOME MEASURES: Serial clinical evaluation and bicycle exercise tests (pre-operatively, at 6 months, and at 1 and 5 years). Quantitative coronary angiography pre-operatively and 5 years after the surgery. RESULTS: Subjects with patent grafts had fewer angina pectoris symptoms at the 5-year follow-up (24 vs. 52%, P = 0.001) and were treated less frequently with long-acting nitrates (3 vs. 15%, P = 0.037) than subjects with graft occlusions. Fewer of them were in classes II-III of the functional classification of the Canadian Cardiovascular Society (39 vs. 74%, P = 0.001). The exercise test was interrupted less often because of chest pain (23 vs. 41%, P = 0.03) and improvement in exercise test variables during the follow-up period was significantly greater in subjects with patent grafts (P<0.002). Amongst patients without graft occlusions, those with new > or =50% diameter stenoses in coronary arteries were more often in functional classes II-III (59 vs. 32%, P = 0.03) than those without new stenoses, but the groups were similar with respect to angina pectoris and exercise tests variables. In patients with graft occlusions, those with and without new > or =50% diameter stenoses were similar with respect to functional class, angina pectoris and exercise test variables. CONCLUSIONS: Angina pectoris and impairment of physical capacity 5 years after coronary artery bypass grafting are mainly due to occlusion of bypass grafts and not to progression of atherosclerosis in coronary arteries. PMID- 10363757 TI - Munchausen syndrome. PMID- 10363759 TI - Magnetic fields produced by single motor units in human skeletal muscles. AB - OBJECTIVE: To develop new non-invasive ways of analyzing human skeletal muscle function, biomagnetic measurement was applied to the vastus lateralis and vastus medialis of 3 healthy adult males using a 64 channel superconducting quantum interference device system. METHODS: Discharges from single motor units were detected by simultaneously recorded surface electromyography. Magnetic signals were averaged 64-158 times at zero-crossing timing in the surface electromyographic signal. RESULTS: Six motor units detected in the 3 subjects produced large magnetic fields with peak-to-peak amplitudes of 1-2 pT. Magnetomyographic isofield maps showed current sources arising from motor endplate regions and propagating in directions opposite to fiber ends. The absolute intensity of current moments in muscle fibers within motor units was estimated based on dipole fitting. The estimated moment was 23.9-114.3 nAm for repolarization dipole. Dividing these moment values by the typical dipole moment of 0.286 nAm in a single muscle fiber, the number of muscle fibers in motor unit was estimated to be 84-400. CONCLUSIONS: Magnetic recording may provide a new non invasive way of analyzing and diagnosing human muscle function. PMID- 10363758 TI - Near ultraviolet radiation elicits visual evoked potentials in children. AB - OBJECTIVE: Ultraviolet radiation can be transmitted through the ocular media, as well as stimulate the retina, in some invertebrate, vertebrate and mammalian species. This study sought to determine if near ultraviolet radiation (UV-A) can elicit visual evoked potentials (VEPs) in young humans. METHODS: VEP responses to 10 nm half-peak bandwidths of 340, 360 and 500 nm stimuli were measured in 8 children aged 7-10 years. Each VEP was based on an averaged response to 200 flashes and was recorded using a sensitivity of 250 microV (full scale) with the International 10-20 electrode placements Fz, O1, Oz, O2, and A1. Peak latencies (ms) were measured for the second negative peak, N2, third positive peak, P3, and third negative peak, N3. The amplitude (microV) between N2 and P3 was also measured. RESULTS: Each child demonstrated a VEP response to both visible and UV A stimuli. Most VEP parameters relative to the 340 and 360 nm stimuli (P < 0.05 to P < 0.001) were significantly different from the VEP responses to the 500 nm stimulus. CONCLUSION: The results indicate that the near ultraviolet stimuli were indeed visible to the young human eye. PMID- 10363760 TI - Vector analysis of visual evoked potentials elicited by pattern reversal and photic stimuli. AB - OBJECTIVE: The 2D VEPs to pattern reversal (PR) and LED goggle were studied in order to obtain a stable parameter for the functional assessing of posterior visual pathways regardless of the stimulus type used. DESIGN AND METHODS: Apex c latency, bc segment amplitude (V), and bc vector orientation angle (theta) in voltage space were computed from VEPs recorded in 50 normal human beings and two patients with left posterior brain lesions, in an orthogonal Fpz-Oz and T3-T4 montage and displayed as a two channel Lissajous' trajectory. The effects of stimulus type and stimulated eye were analyzed in the normal group by a two-way ANOVA. RESULTS: The stimulated eye had no effect on any parameter. Apex c latency was slightly longer, and V was greater and more variable in the responses to goggle stimuli, but there was no significant difference in theta, oriented to mid occipital scalp with very low variability for both stimulus types. The patients showed significant deviations of theta towards the affected hemisphere. CONCLUSIONS: The bc vector orientation (theta) is a stable parameter for the evaluation of the posterior visual pathways using both pattern reversal and LED stimuli, specially the latter, useful in unconscious or uncooperative patients. PMID- 10363761 TI - Movement-related cortical potentials associated with voluntary relaxation of foot muscles. AB - OBJECTIVE: In our previous study of movement-related cortical potential (MRCP) in association with the voluntary relaxation of the hand muscle, Bereitschaftspotential (BP) was maximal at the vertex and symmetrically distributed, and Negative Slope (NS') was maximal over the contralateral central region. In order to clarify the generator sources of MRCP with voluntary muscle relaxation, we recorded MRCP in association with voluntary relaxation of the foot. METHODS: MRCP in association with plantar flexion of the foot caused by voluntary relaxation of the tibialis anterior muscle was recorded in 10 normal subjects. RESULTS: The BP started at about 1.7 s before the onset of the muscle relaxation, followed by NS' starting at about 650 ms before it. Both were maximal at the vertex and symmetrically distributed. There was no additional EEG activity in the lateral frontal areas, which are presumably located over the primary negative motor areas (PNMA). CONCLUSIONS: It is concluded that the voluntary muscle relaxation, similarly to the voluntary muscle contraction, involves the cortical preparatory activity at least in the primary motor area (M1) and probably the supplementary motor areas (SMAs). There is no evidence to suggest that the PNMA is also active prior to the voluntary muscle relaxation. PMID- 10363762 TI - Spike detection II: automatic, perception-based detection and clustering. AB - OBJECTIVES: We developed perception-based spike detection and clustering algorithms. METHODS: The detection algorithm employs a novel, multiple monotonic neural network (MMNN). It is tested on two short-duration EEG databases containing 2400 spikes from 50 epilepsy patients and 10 control subjects. Previous studies are compared for database difficulty and reliability and algorithm accuracy. Automatic grouping of spikes via hierarchical clustering (using topology and morphology) is visually compared with hand marked grouping on a single record. RESULTS: The MMNN algorithm is found to operate close to the ability of a human expert while alleviating problems related to overtraining. The hierarchical and hand marked spike groupings are found to be strikingly similar. CONCLUSIONS: An automatic detection algorithm need not be as accurate as a human expert to be clinically useful. A user interface that allows the neurologist to quickly delete artifacts and determine whether there are multiple spike generators is sufficient. PMID- 10363763 TI - Somatic innervation of the human bulbocavernosus muscle. AB - OBJECTIVE: To demonstrate the somatic reflex innervation of the bulbocavernosus muscle (BCM), the principal muscle for ejaculation. METHODS: Genitourinary electrodiagnostic testing utilizing modifications of the standard bulbocavernosus reflex was performed in 13 healthy male volunteers ages 20-43. RESULTS: Bulbocavernosus muscle contraction was elicited by stimulation of the dorsal nerve of the penis, from both the penile skin and from the anterior urethra, and following stimulation of the perineal nerve. Latencies were variable depending on the point of stimulation. CONCLUSIONS: All 3 afferent pathways synapse on pudendal motoneurons in the conus medullaris, and provide for peripheral reflex control of BCM contractions. Based on the latencies of the urethral evoked responses, urethral innervation differs from penile shaft innervation, each having a distinct population of the dorsal nerve of the penis (DNP) fibers. The presence of an electrically-defined pathway from the anterior urethra to the BCM suggests that somatic afferents from the anterior urethra are involved with the ejaculatory reflex. These somatic reflexes are components of normal ejaculatory function. The findings contribute to understanding the neurophysiology of ejaculation, and may be applicable to the evaluation of ejaculatory disorders. PMID- 10363764 TI - Transient epileptic foci associated with intracranial hemorrhage in patients with subdural and epidural electrode placement. AB - We report two cases of transient epileptic foci in humans associated with placement of intracranial electrodes. The abnormalities consisted of restricted areas of active, almost continuous, rhythmic spiking, intermittently evolving into electrographic seizure activity, which resolved spontaneously within 3-4 days. The first occurred after placement of a subdural electrode grid and the second following insertion of epidural peg electrodes. Neuroimaging demonstrated a small subdural hematoma overlying the grid and a focal intraparenchymal hemorrhage underlying the affected epidural electrodes. The insertion of intracranial electrodes may be complicated by the induction of transient epileptic foci unrelated to a patient's typical epileptic generator. PMID- 10363765 TI - Anticipation and execution of a simple reading task enhance corticospinal excitability. AB - OBJECTIVE: Electromyographic responses (EMG) evoked in the right hand by transcranial magnetic stimulation (TMS) of the left motor cortex are enhanced during continuous reading. This enhancement is the result of increased excitability of the motor cortex. We proposed that anticipation and reading of single words would also enhance corticospinal excitability. We studied the temporal course of corticospinal excitability changes following left and right hemisphere TMS. METHODS: Ten normal volunteers were studied. A warning stimulus (S1) was followed by an imperative stimulus (S2) whereupon a word was presented. Subjects responded by reading the word aloud or reading it silently. In other conditions, no word was displayed and the subjects responded to S2 by saying the word 'Cat', pursing their lips, or doing nothing. EMG was recorded over the contralateral hand following a TMS pulse over the motor cortex during and after the S1-S2 period. RESULTS: Enhancement of EMG amplitudes was significantly greater following left hemisphere TMS. The enhancement in the S1-S2 period and that following S2 had a time course similar to several event-related brain potentials. CONCLUSIONS: There may be a common mechanism underlying both corticospinal excitability and the contingent negative variation, readiness potential and N400. PMID- 10363766 TI - Are there discrete distal-proximal representations of the index finger and palm in the human somatosensory cortex? A neuromagnetic study. AB - OBJECTIVE: The distal-proximal representations of the finger and palm in the first somatosensory cortex (SI) were studied in humans. METHODS: Somatosensory evoked magnetic fields (SEFs) from 11 subjects were measured, following mechanical stimulation of the skin by using a 122 channel whole head SQUID system. Sensory stimulus comprising of a 10 ms vibration at the frequency of 200 Hz was delivered to 6 successive sites in 3 cm increments, along the distal proximal direction over the volar surface of the right index finger and palm. Using a single dipole model, the sources of the magnetic fields were estimated and mapped onto magnetic resonance images of each subject. ANOVA was used for statistics. RESULTS: Source localization was determined on the main peak (M50) of the SEFs. All of the sources were located in the area 3b of SI. Contrary to the well-defined distal-proximal representations in the hand area of simian SI cortex, there was no statistically significant differences between the locations of the dipoles in human SI cortex evoked by stimulation of different sites. CONCLUSION: The result, however, should be interpreted with caution, because it cannot be denied that the spatial separation of sources in the distal-proximal somatotopy is beyond the resolving capacity of magnetoencephalography (MEG). In addition, at variance with the discrete distal-proximal gradient in the mechanoreceptor density, there was no statistically significant differences between the signal strengths of the dipoles for stimulation of the different locations. PMID- 10363767 TI - The significance of excessive rhythmic alpha and/or theta frequency activity in the EEG of the neonate. AB - OBJECTIVE: The goal of this study was to determine the significance of excessive rhythmic alpha and/or theta frequency activity in neonatal EEGs. METHODS: The EEGs of 963 neonates, 26-44 weeks conceptional age (CA), performed during the years 1992-1994 at the Texas Children's Hospital, Houston, Texas, were reviewed for the presence of excessive rhythmic alpha and/or theta frequency activity. Cases in which such activity was identified were further characterized by the presence or absence of other EEG abnormalities. The medical records of these patients and a group of control infants with normal EEGs were reviewed to identify associated pathological conditions. RESULTS: Forty patients were identified whose EEGs revealed such activity. The CA of these patients ranged from 37 to 44 weeks. A variety of pathological conditions were seen in these patients, most commonly congenital heart disease, congenital brain anomalies and hypoxia. These conditions were not seen in the control group of infants. Twenty patients had received CNS-active drugs. The EEGs of 32 patients revealed additional abnormalities, most commonly multifocal sharp waves and episodes of voltage attenuation during slow-wave sleep. CONCLUSIONS: The results indicate that excessive rhythmic alpha and/or theta frequency activity is an abnormal finding in the newborn's EEG. PMID- 10363768 TI - Transcranial magnetic stimulation of the central and peripheral motor pathways to the lingual muscles in cats. AB - OBJECTIVE: We have assessed a technique to stimulate the intracranial hypoglossal nerve with ease and reproducibility by using magnetic coils (MCs) and to detect a reliable site of excitation in animal experiments in order to establish a method to evaluate the motor pathway to lingual muscles. METHODS: We recorded the motor responses from the lingual muscles of 5 adult cats under general anesthesia by magnetic and electrical stimulation of the intracranial hypoglossal nerves. Figure of 8 and round MCs were used to investigate the optimal position and direction to evoke the motor responses. RESULTS: The round MC was useful for cortical stimulation. The figure of 8 coil, positioned in the back of the head of the examined side, parallel to the cervical spine, was essential for stimulation of the intracranial hypoglossal nerve. Analysis of the latencies, and the observation that the motor responses disappeared after transection of the nerves at the exit of the hypoglossal canal, demonstrated that the site of the excitation is at the exit of the hypoglossal canal. CONCLUSION: Magnetic stimulation using a figure of 8 coil can elicit tongue motor responses with ease and reliable reproducibility, stimulating the hypoglossal nerve at the exit of the hypoglossal canal. PMID- 10363769 TI - Task modulation of brain activity related to familiar and unfamiliar face processing: an ERP study. AB - In order to investigate stimulus-related and task-related electrophysiological activity relevant for face processing, event-related potentials (ERPs) from 58 electrodes at standard EEG sites were recorded while subjects performed a simple visual discrimination (control) task, in addition to various face processing tasks: recognition of previously learned faces and gender decision on familiar and unfamiliar faces. Three electrophysiological components or dipolar complex were recorded in all subjects: an occipital early component (P1, around 110 ms); a vertex positive potential (VPP; around 158 ms) which appeared to be specific to faces; and a negative central component, N2 (around 230 ms). Parametric analysis and source localization were applied to these components by means of a single subject analysis methodology. No effect of familiarity was observed on any of these early component. While the VPP appears to be independent of the kind of processing performed, face task modulations of the early P1 and the N2 were observed, with a higher amplitude for the recognition than for the gender discrimination task. An attentional modulation of early visual areas is proposed for the first effect (P1 modulation), while the N2 seems to be related to general visual memory processing. This study strongly suggests that the VPP reflects an early visual stage of face processing in the fusiform gyrus that is strictly stimulus-related and independent of familiarity. It also shows that source localization algorithms may give reliable solutions on single subject averages for early visual components despite high inter-subject variability of the surface characteristics of ERPs. PMID- 10363771 TI - EEG coherency II: experimental comparisons of multiple measures. AB - OBJECTIVE: A concentric spheres model was used in an earlier paper to estimate the effects of volume conduction, reference electrode and spatial filtering on different EEG coherence measures. EEG data are used here to verify theoretical predictions. METHODS: Three EEG data sets were: (1) 64 channel, recorded during 7 alternating periods of resting and mental calculation. (2) 128 channel, for comparison of eyes open versus eyes closed coherence. (3) 128 channel, recorded during deep sleep (stages 3 and 4) and REM. RESULTS: The directions of large scale (lobeal) coherency changes between brain states are relatively independent of coherence measure. However, coherence between specific electrode pairs is sensitive to method and frequency. Average reference and digitally linked mastoids provide reasonable semi-quantitative estimates of large-scale neocortical source coherence. Close bipolar, Laplacian, and dura image methods remove most reference electrode and volume conduction distortion, but may underestimate coherence by spatial filtering. CONCLUSION: Each EEG coherence method has its own potential sources of error and provides coherence estimates for different neural population sizes located in different locations. Thus, studies of coherence and brain state should include several different kinds of estimates to take full advantage of information in recorded signals. PMID- 10363770 TI - Auditory and visual P300 topography from a 3 stimulus paradigm. AB - OBJECTIVE: The P300 event-related brain potential (ERP) was elicited with auditory and visual stimuli in a separate session of a 3 stimulus oddball paradigm, and the scalp topography was assessed with 15 electrode locations. METHODS: Target (0.10), standard (0.80), and infrequent non-target (0.10) stimuli in the auditory task were 2000, 1000 and 500 Hz tone, and in the visual task, 'X', 'O', and 'H', respectively. The stimuli were presented in a random series, once every 2 s, and participants responded only to the target (N = 12). RESULTS: Target stimuli elicited larger P300 components than non-target did in both stimulus modalities. For both target and non-target stimuli, P300 amplitude was larger and latency longer for the visual compared with the auditory stimulus. Analysis of normalized P300 amplitude data indicated that the target and non target P300s from both modalities had identical topography. CONCLUSION: The findings suggest that both target and non-target stimuli in 3 stimulus oddball paradigm elicited the same type of P300 (P3b) for both stimulus modalities. PMID- 10363772 TI - Physiological studies of the corticomotor projection to the hand after subcortical stroke. AB - OBJECTIVE: The mechanisms which lead to recovery of motor function after a stroke are poorly understood. Functional reorganization of cortical motor centres is thought to be one of the factors which may contribute to recovery. We have investigated the extent of reorganization which occurs at the level of the primary motor cortex after a lesion of the corticospinal pathway. METHODS: Transcranial magnetic stimulation was used to map the topography of the primary corticomotor projection to the abductor pollicis brevis muscle and study changes in cortical motor thresholds and corticospinal conduction in a group of 20 subjects with subcortical infarcts of varying duration (1 week to 15 years) and varying degrees of motor deficit. RESULTS: There was a broad correlation between motor evoked potential (MEP) amplitude and motor thresholds on the one hand and the severity of motor deficit and site and extent of the lesion on the other. Shifts in the cortical motor maps were found in both early and late cases, irrespective of the site of the lesion, but were more frequent in the longer standing cases. Shifts were usually along the mediolateral axis but anteroposterior shifts were found in some late cases. CONCLUSION: Our findings indicate that there is functional reorganization of the corticomotor projection in subjects who regain a degree of motor control following a subcortical lesion sparing the motor cortex. PMID- 10363773 TI - Effect of stimulation of an upper limb on motor evoked potentials in lower limb muscles to transcranial magnetic stimulation in normal subjects and patients with thalamic infarction. AB - The effects of conditioning stimulation of an upper limb on motor evoked potentials (MEPs) of relaxed muscles in both lower limbs were studied in 7 normal subjects and two patients with left thalamic infarction. A possible mechanism for the Jendrassik maneuver (JM) is that induced proprioceptive input ascends supraspinally to facilitate the descending volleys. In order to mimic the JM with a more controlled influence, we used an electrical conditioning (C) stimulation (4 times sensory threshold) delivered to the left index finger preceding the transcranial (T) magnetic stimulation at C-T intervals of 0-200 ms. The MEP facilitation of bilateral tibialis anterior (TA) and gastrocnemius medialis (GC) was within C-T 70-110 ms. The peak facilitation was at C-T 80 ms for ipsilateral TA (309%) and GC (405%) and at C-T 90 ms for contralateral TA (207%) and GC (283%). In the two thalamic infarction patients with right-sided sensory loss, the facilitation did not occur when the conditioning stimulation was delivered to the affected index finger. Therefore, it is likely that the peripheral volley must be transmitted supraspinally to facilitate MEPs of the lower limbs. This method for studying sensory facilitation is more quantitative and reproducible than the JM and technically better than other previously described methods for somatosensory conditioning. PMID- 10363774 TI - Abnormal contingent negative variation in writer's cramp. AB - OBJECTIVE: To investigate the physiological abnormality in writer's cramp, a focal dystonia which specifically affects writing. METHODS: We recorded brain potentials that precede hand and neck movements (contingent negative variation or CNV) in 11 patients and 11 age-matched normal subjects. A 1000 Hz tone burst (S1) was delivered to the right or left ear in random sequence, and 2 s after, a 2000 Hz tone burst (S2) was delivered to both ears simultaneously. For the response task to S2, the subjects were instructed to extend their fingers ipsilateral to the ear to which S1 was given in one experiment or to rotate the head to the side of the S1 presentation in another. All the patients had symptoms in the right hand only, and performed both tasks normally. CNV amplitudes were compared between normals and patients using unpaired t test. RESULTS: They showed normal CNV for neck movement but significantly decreased CNV amplitudes for movements both in the affected and unaffected hands. CONCLUSIONS: Our findings suggest that motor programming is specifically abnormal for the affected body part, including the asymptomatic contralateral limb, and that the clinical symptom may result from a deficient compensatory mechanism for abnormal motor programs or subroutines. PMID- 10363775 TI - Cerebral correlates of hemispheric lateralization during a pitch discrimination task: an ERP study in dichotic situation. AB - OBJECTIVE: Electrophysiological correlates of perceptual asymmetry for dichotic pitch discrimination were investigated in 12 right-handed volunteers, whose dichotic listening performances attested the classical 'right ear advantage' in a verbal discrimination task. METHODS: Event related potentials (ERPs), elicited by dichotic and binaural pairs of tones applied in a classical oddball paradigm including right ear targets, left ear targets and binaural targets (5% occurrence each) were recorded from medial and lateral scalp locations. Latencies and baseline to peak amplitudes were measured for P1, N1, P2, N2 and P3 components. RESULTS: ERPs recorded in response to dichotic (compared with binaural) target pairs, exhibited delayed latencies for N2 and P3, correlated with prolonged RTs, probably linked to greater difficulty in identification of the target. They also displayed enhanced N1 and P2 voltages, which may reflect the simultaneous activation of two different populations of neurons in the auditory cortical areas. We observed specific lateralization effects for pitch discrimination with a left ear advantage on latency of early components. CONCLUSIONS: Together with amplitude asymmetries in the N2 component, the findings bring strong electrophysiological support to Kimura's structural model for dichotic perceptions with a right hemisphere prevalence in a pitch discrimination task. PMID- 10363776 TI - Increased gamma-range activity in human sensorimotor cortex during performance of visuomotor tasks. AB - OBJECTIVE: We documented changes in spectral power of human electrocorticograms (ECoG) during performance of sensorimotor tasks. METHODS: In 6 human subjects, ECoGs were recorded simultaneously from 14 subdural cortical sites in forearm sensorimotor cortex. The subjects performed 3 visuomotor tasks: tracking a moving visual target with a joystick-controlled cursor, threading pieces of tubing, and pinching the fingers sequentially against the thumb. Control conditions consisted of passive resting and active extension of the wrist. For each site the spectral power of the ECoG during these behaviors was computed for 5 10 Hz ranges between 10 and 60 Hz. RESULTS: All subjects showed power decreases in the range of 11-20 Hz and power increases in the 31-60 Hz range during performance of the visuomotor tasks, at sites in forearm sensorimotor cortex and adjacent areas. Simple wrist movements often produced little change in power. Three subjects showed episodes of explicit gamma oscillations during the visuomotor tasks. Different sites showed increases in gamma-range power for different tasks, indicating that the spatial distribution of the gamma activity is specific to the tasks. Cross spectra showed that gamma activity could become synchronized between separate sites during particular tasks. CONCLUSIONS: Synchronized gamma-range activity in human sensorimotor cortex increases with performance of manipulative visuomotor tasks, supporting the hypothesis that coherent gamma oscillations may play a role in sensorimotor integration or attention. PMID- 10363777 TI - Distributed source imaging of alpha activity using a maximum entropy principle. AB - OBJECTIVE: We present a method based on the distributed dipole source model to localize sources of spontaneous human brain activity, such as the alpha rhythm. The proposed method relies on the generalized maximum entropy principle and is implemented in frequency-domain. METHODS: Several computer simulation studies of synchronous and asynchronous distributed dipole sources were carried out to test the validity of the method. The method was also applied to spontaneous electroencephalographic (EEG) recordings from human subjects to estimate the sources of alpha activity. The locations of these sources were registered with actual magnetic resonance images for anatomical visualization. RESULTS: The simulation studies suggest the validity of the proposed method and its capability to detect distributed and fairly deep synchronous dipole sources. Results of human studies with 6 subjects suggest that the generators of alpha rhythm are mainly concentrated over the posterior regions of the cortex. CONCLUSION: The proposed distributed source imaging method is a promising technique for localizing rhythmic brain activity. PMID- 10363778 TI - On the focal nature of inhibition and facilitation in the human motor cortex. AB - OBJECTIVE: To find the area of the human cortex from which inhibition and facilitation of corticospinal neurons could be obtained. METHODS: A patient with seizures had an array of 64 electrodes placed over the left fronto-temporal cortex. The motor evoked potential (MEP) elicited by stimulating through one pair of electrodes was conditioned by stimuli that were subthreshold for a MEP given through adjacent pairs of electrodes. RESULTS: The MEP recorded over the right abductor pollicis brevis produced by stimulating over the hand area of the left cortex could be inhibited (at intervals less than 5 ms) and facilitated (at intervals greater than 5 ms) by subthreshold conditioning stimuli delivered through neighbouring pairs of electrodes. The inhibition and facilitation were only obtained when the conditioning stimuli were delivered within 1-2 cm of the test site. The sites producing inhibition and facilitation were not identical. Conditioning stimuli over the face area did not inhibit the MEP produced by stimulating the hand area or vice versa. CONCLUSION: The inhibition and facilitation of corticospinal neurons projecting to a given muscle arise from small areas close to those corticospinal neurons. PMID- 10363779 TI - Normative ranges by curve fitting to raw data. AB - This study proposes a technique for finding the frequency distributions (FDs) of the median motor distal latencies (MMDL) of healthy and of disease subjects by mathematically analyzing the combined frequency distribution (CFD) of all the subjects that were recorded in an EMG Lab, from the numbers themselves, without any reference to clinical data. The CFD is the algebraic summation of the healthy group FD and the disease-group FD. Although it is impossible to visually detect these FDs within the CFD numerical values or graphic representation, the hypothesis is that it can be done analytically. Three analytical ways were tested and showed satisfactory results. Also, this study shows the statistics of the overlap between health and disease, i.e. the prediction value of each MMDL, statistics that are of utmost importance, and can not be generated in any other way. PMID- 10363780 TI - Comparison of endogenous event-related potentials in attend and non-attend conditions: latency changes with normal aging. AB - OBJECTIVE: Endogenous event-related potential (ERP) components have been observed under both attend and non-attend conditions, but it appears that at least some of the attend and non-attend components are functionally and topographically distinct. Also, under active task conditions, motivational and attentional variations may modulate the amplitude of the ERP. These various effects of attention on the ERP can complicate comparisons of the ERPs of normal subjects with the ERPs of clinical subjects, who may have reduced attentional capabilities. The experiment reported here sought to develop a non-task paradigm that reliably produces the same ERP components typically seen under task conditions. METHODS: Using rare auditory stimuli that were discrepant from the frequent stimuli both in frequency and intensity, stimuli were presented under non-attend instructions and under instruction to count the rare stimuli. The ERPs in these two conditions were compared with ERPs in a standard oddball paradigm which used stimulus parameters comparable to those of most previous experiments on ERPs in aging. Fifty subjects, ranging in age from 20 to 77, participated. RESULTS: The ERPs to the DISCREPANT oddball stimuli under non-task conditions were similar in scalp distribution to the ERPs to the same stimuli in the ATTEND condition and to the ERPs in the STANDARD/ATTEND condition. Furthermore, there were no significant differences in the age-related increase in ERP latencies among the DISCREPANT/IGNORE, the DISCREPANT/ATTEND, and the STANDARD/ATTEND conditions. CONCLUSION: The results indicate that increases in ERP latencies with aging can be assessed in the absence of task requirements, and that the paradigm described here may prove useful in investigating cognitive processing speed in clinical populations. PMID- 10363781 TI - Modulation of intracortical excitability for different muscles in the upper extremity: paired magnetic stimulation study with focal versus non-focal coils. AB - OBJECTIVE: Intracortical excitability was studied for 4 muscles in the upper extremity by paired transcranial magnetic stimulation on the motor cortex, using focal and non-focal coils. METHODS: Surface EMG responses of two hand and two forearm muscles were simultaneously recorded in 13 healthy subjects, after delivering paired stimuli at interstimulus intervals (ISIs) of 1-50 ms using circular and figure-of-8 (focal) coils. The intensities of conditioning and test stimuli were submotor and supramotor thresholds, respectively. RESULTS: Paired stimulation using a circular coil showed constant inhibition at 2 ms ISI for all muscles, but not at 5 ms ISI, and induced facilitation at 10 ms ISI. The results using a focal coil were similar to those with a circular coil at all ISIs except at 5 ms ISI, where the former showed inhibition. At 20 and 30 ms ISIs, there was interindividual variability for both coils, some subjects showing inhibition and others facilitation. CONCLUSIONS: The difference of the inhibition at 5 ms ISI between using circular and focal coils could be attributed to the cortical mechanism. The inhibitory effect at 2 ms ISI, consistently observed for the 4 muscles with both types of coil, could be easily applied to assess the inhibitory intracortical function in patients with neurological diseases. PMID- 10363782 TI - Hematopoietic stem cells and aging. AB - The question of whether hematopoietic stem cells are altered in aging has been the subject of considerable controversy for over two decades. The substantial advancement of knowledge on hematopoietic stem cells and developmental hematology in the last few years has reopened this issue for critical analysis. Dynamic changes have been noted regarding the anatomic site and the function of hematopoietic cells, from the early embryo to old age. Whereas basal hematopoietic potential is maintained in aging. the capacity for recovery from hematological stress and for stem cell self-renewal appears to decline gradually. A distinction is thus made between the steady-state hematopoiesis in aging and the developmental potential of stem cells. The establishment of proper tools to identify and to study purified stem cells and committed cell populations offers a direct approach to further elucidate aging across the axis from primitive stem cells to the mature blood cells. The present article represents a brief review of this area. PMID- 10363783 TI - Transgenic mouse models of muscle aging. AB - In the last decade transgenic animals have been become a powerful and exciting research model to study the molecular mechanisms underlying the cellular and physiological processes such as cell growth, differentiation, apoptosis, and the regulation of specific gene expression. In the context of skeletal muscle development, transgenic mice and gene-targeting approaches have led to the definition of specific roles for Muscle Regulatory Factors (MRFs) during embryogenesis, although less is known about the molecular mechanism underlying skeletal muscle aging. Recent studies using specific models of transgenic mice have added new insights into the muscle aging process, providing a baseline for designing appropriate strategies to attenuate or to reverse the cumulative effects of aging. In this review we discuss some of the transgenic models currently available to address the molecular mechanisms of skeletal muscle senescence. Given the complexity of the aging process, this review should be regarded as a presentation of works in progress rather than a comprehensive description of muscle aging. PMID- 10363784 TI - A mild stress, hypergravity exposure, postpones behavioral aging in Drosophila melanogaster. AB - Flies were submitted to two weeks of hypergravity in a centrifuge (3 or 5 g), starting at the second day of imaginal life, and their behavior (spontaneous locomotor activity, patterns of movement, and climbing activity) was observed from removal of the centrifuge to an older age; the usual effects of age on these behaviors were generally observed. Hypergravity-kept flies had worse behavioral scores on removal of centrifuge than those always kept at 1 g. When they aged, they got either similar or better scores than 1 g flies, which indicates that their behavioral aging may be slower. These results show that a mild stress such as hypergravity, which has been previously shown to increase the longevity of males and resistance to heat shock in both sexes, is an environmental manipulation postponing aging in flies. PMID- 10363785 TI - Extended longevity lines of Drosophila melanogaster: abundance of yolk protein gene mRNA in fat body and ovary. AB - Lines of Drosophila melanogaster selected for late-life female reproduction typically exhibit correlated responses of reduced early fecundity and increased longevity. This relationship suggests a tradeoff between reproductive effort and somatic maintenance, which in turn, underlies some evolutionary theories of senescence. The mechanistic basis of the apparent tradeoff between increased longevity and reduced early-age fecundity has remained obscure. The present manuscript addresses the issues of whether the reduced early-age fecundity in selected lines corresponds to reduced yolk-protein mRNA production, and whether long-lived flies exhibit somatic maintenance in terms of relatively reduced yolk protein mRNA production in the fat body. Yolk protein is one of the most abundant proteins used for female reproduction. By comparing a set of lines selected for late life reproduction with the corresponding control lines, we show that that yolk-protein gene mRNA relative abundance during the first four days posteclosion did not correspond to reduced early-life fecundity in the selected lines. In D. melanogaster, yolk protein is produced in the fat body and ovarian follicle cells. On the fourth day posteclosion, relatively more yolk-protein gene mRNA was present in the fat body. On day 1 posteclosion, supplemental yeast did not alter relative yolk-protein gene mRNA abundance. However, on day 4 posteclosion, supplemental yeast stimulated yolk-protein gene mRNA production in the fat body, which suggests an underlying mechanism for the nutrition-based phenotypic plasticity of fecundity previously documented in these lines. On medium without supplemental yeast, the relatively low abundance of fat body yolk-protein gene mRNA in the selected lines on day 4 posteclosion corresponds to a prediction derived from the disposable soma theory. PMID- 10363786 TI - Activity of superoxide dismutase and catalase in the bean weevil (Acanthoscelides obtectus) selected for postponed senescence. AB - Relationship of superoxide dismutase and catalase activities and aging were tested using bean weevil lines selected for postponed senescence. The beetles of different age (young and old) and mating status (virgin and mated) from the extended longevity lines were compared with their counterparts derived from the short-lived lines for activities of SOD and catalase. The old beetles from the long-lived lines had statistically significant higher activity of SOD than their controls. Although we did not find a significant effect of catalase on longevity, beetles originating from both types of lines exhibited an increased catalase activity during mating processes. In addition, we did observe an increased activity of catalase in one-day-old beetles of the short-lived lines relative to the same-aged individuals of the long-lived lines. PMID- 10363787 TI - Age-related impairment of p56lck and ZAP-70 activities in human T lymphocytes activated through the TcR/CD3 complex. AB - Cellular immune responses decrease with aging. Lymphocytes of aged individuals do not perform as well as cells from young subjects in a number of in vitro assays including cell proliferation, cytokine production, and protection against apoptosis. Here, we have tested the hypothesis that a decrease in T cell responses in tymphocytes from elderly subjects could parallel a decrease in the activity of protein tyrosine kinases (PTK) associated with signal transduction in T lymphocytes. We report that anti-CD3-triggered T lymphocyte proliferation was significantly decreased in T lymphocytes from elderly subjects, but the decrease was not due to an alteration of the percentage or mean fluorescence intensities of CD3, CD4, and CD45. Of significance, the activities of p56lck and ZAP-70 in vitro were significantly decreased in T lymphocytes from elderly subjects compared to young individuals. However, the level of expression of the two kinases did not change with aging. The activity of p59fyn did not show changes with aging, suggesting that p59fyn did not compensate for the decreased activity of p56lck. We also found that the extent of tyrosine phosphorylation of the adaptor protein p95vav was similar in activated T lymphocytes from elderly and young subjects. Our results suggest that the altered cellular immune responses observed in T lymphocytes with aging may be the result, at least in part, of an alteration in early events associated with signal transduction through the TcR/CD3 complex that translates into decreased activities of p56lck and ZAP-70. Impairment in the activities of these twokey components of T cell signaling may contribute to reduced immune functions associated with aging. PMID- 10363788 TI - Early lymphocyte activation in elderly humans: impaired T and T-dependent B cell responses. AB - Immunosenescence is characterized by an increase in autoantibody production. Because both T and B cell stimulation are key events for producing antibodies, we investigated early T and B cell activation by means of CD23 and CD40L (two very early activation antigens). PBMC from elderly humans (EH) were studied following culture with either medium, anti-CD3mAb, rIL-4, or PMA + ionomycin. CD23 expression on elderly B cells after anti-CD3 challenge of PBMC, a reflect of T dependent B cell activation, was clearly defective. Conversely, CD23 expression on EH B cells following activation with soluble factors as rIL-4 was preserved. CD40L expression was also impaired in EH T cells following anti-CD3 challenge. However, activation by means of PMA and/or ionomycin was preserved both in T cells (CD40L expression) and in B cells (CD23 expression). These results indicate that a defective T-dependent B cell activation related to defective T cell activation located between surface membrane and PKC/ionomycin function is an intrinsic characteristic of immunosenescence. We have not found intrinsic B-cell defects, and we conclude that the characteristically impaired early B cell activation in EH is mostly due to T cell defects. PMID- 10363789 TI - Age-related alteration of cytokine production profile by T cell subsets in mice: a flow cytometric study. AB - Spleen cells from young and old C57BL/6 mice were stimulated with a combination of anti-CD3 and anti-CD28 antibodies, and the profile of cytokine production was examined by two different methods; the concentrations of cytokines as measured by ELISA, and identification of cytokine-positive cells by flow cytometry. The ELISA method revealed that IL-2 production by spleen cells after stimulation was significantly lower in the old mice compared to the young mice. while IFN-gamma production was the reverse. The flow cytometric analysis showed that the percentage of IL-2 positive cells in spleen cells after the stimulation was significantly lower in the older mice than in the young mice, and vice versa for the percentage of IFN-gamma-positive cells. Regarding the T cell subsets, CD4+ T cells were a major source of IL-2 in both the young and old mice. IL-2-positive cells in both CD4+ and CD8+ T cells showed a significant decrease with age. On the contrary, CDX T cells were the major source of IFN-gamma. An age-related increase of IFN-gamma positive cells was observed in both CD4+ and CD8+ T cells. CD4 T cells were the major source of IL-4, and the percentage of IL-4-positive CD4+ T cells also increased with age, although the level of IL-4 production was modest in C57BL/6 mice compared with IL-2 and IFN-gamma. Such age-related changes of cytokine production are presumed to play an important role in the alteration of immunological capacity with age. PMID- 10363790 TI - Age-related impaired proliferation of peripheral blood mononuclear cells is associated with an increase in both IL-10 and IL-12. AB - Reflective of age-associated decline in immune function among elderly individuals is a decrease in in vitro T cell proliferative ability. Impaired T cell proliferation in the elderly may result from disruption of the well-balanced network of regulatory cytokines produced during an immune response. The purpose of this study was to identify age-related changes in the production of interleukin (IL)-10 and IL-12, and to determine whether in vitro T cell proliferation can be enhanced in the elderly by modulation of these two key cytokines. The superantigen Staphyloccocus entertoxin B (SEB) was used to stimulate proliferation and IL-10 and IL-12 production in peripheral blood mononuclear cells (PBMC) in vitro. Proliferation was determined by standard tritiated thymidine uptake. Cytokine levels in culture supernatants were measured by ELISA. We observed impaired SEB-induced proliferation of PBMC in the elderly that is comparable to that seen with the polyclonal mitogen Con A. This age related decline in proliferation was associated with increased production of both IL-10 and IL-12. Modulation of PBMC proliferative response with either recombinant IL-12 or IL-10-neutralizing antibodies can boost proliferation of elderly PBMC to the levels seen in unmodulated young controls. PMID- 10363791 TI - NK phenotypic markers and IL2 response in NK cells from elderly people. AB - Immunosenescence is a process that primarily affects the T cell compartment of the immune system, although age-associated immunological alterations have also been demonstrated in the NK cell phenotype and function. A significant expansion in the number of NK cells is found in aging. The NK cytotoxic capacity of total peripheral blood lymphocytes is also well preserved, not only in healthy elderly people but also in centenarians. However, NK cell killing of K562 is impaired when considered in a per-cell basis, and this defect is associated with defective signal transduction after activation more than a diminished conjugate formation or killing capacity. We have studied the phenotype of NK cells in elderly donors fulfilling the Senieur criteria. We have also studied the capacity of these cells to be activated by IL2 when different NK cell functions, other than cytotoxicity, are considered. Our results confirm the increased percentage of NK cells in the elderly due to the expansion of the CD56dim subset that also show an altered pattern of activation markers, whereas no differences were found in the CD56bright subset. The response of NK cells to IL2 was found to be impaired when proliferation, expression of CD69, and Ca2+ mobilization were considered, whereas TNF-alpha production was not significantly affected. These results suggest that human NK cells do not escape the aging process, although senescence have a differential effect on distinct NK cell biological functions, ranging from severe to negligible impairment, depending on the parameters considered. PMID- 10363792 TI - Mechanisms of age-related changes in gonadotropin-releasing hormone receptor messenger ribonucleic acid content in the anterior pituitary of male rats. AB - To determine the mechanism(s) of age-related changes in gonadotropin release from pituitary gonadotrophs in male rats, we measured the number of GnRH (gonadotropin releasing hormone) receptor containing cells and expression of GnRH receptor mRNA per cell in the anterior pituitary. An in situ hybridization procedure was performed using young (six months) and old (24-25 months) Wistar rats. An image analysis system was employed for the autoradiographic analysis. The number of pituitary cells increased during aging (approximately 45%, p < 0.01). On the other hand, the number of GnRH receptor mRNA-containing cells decreased (approximately 25%, p < 0.05). The percentage of these cells in old rats decreased to less than a half of that in young animals (p < 0.01). GnRH receptor mRNA per cell in old rats was only 7% lower than in young (p < 0.01). These results suggest that loss of pituitary gonadotroph GnRH receptors and response is primarily due to the loss of gonadotrophs, and that the death mechanism(s) are responsible for decreased stimulation of Gn release during aging. PMID- 10363793 TI - Proliferative disorders of the aging human prostate: involvement of protein hormones and their receptors. AB - The majority of elderly men is affected by benign and malignant diseases of the prostate. Both proliferative disorders, i.e., benign hyperplasia of the prostate (BPH) and prostate cancer (PCa)-which has recently emerged as the most common male malignancy in industrialized countries-seem to be governed by endocrine factors such as sex steroid hormones, but auto/paracrine factors are involved as well. Age-related changes in levels and ratios of endocrine factors as androgens, estrogens, gonadotropins, and prolactin (PRL) and changes in the balance between auto/paracrine growth-stimulatory and growth-inhibitory factors such as insulin like growth factors (IGFs), epidermal growth factor (EGF), nerve growth factor (NGF), IGF-binding proteins (IGFBPs), and transforming growth factor beta (TGFbeta) are meant to be responsible for abnormal prostatic growth. We investigated the existence of putative local regulatory circuits involving the protein hormones, human growth hormone (hGH), human placental lactogen (hPL), and hPRL, and their corresponding receptors in prostatic tissue specimens (transurethral resections of the prostate, TURP; n = 11), in the prostatic cancer cell lines PC3, Du145, LnCap, a virus-transformed BPH cell line (BPH-1), and in a normal healthy prostate by RT-PCRs and highly specific and sensitive immunofluorometric assays (IFMA). Neither hPRL nor hGH was detected at the mRNA or protein levels in prostatic tissue and cell lines, with the exception of 2 of 11 prostatic TURP-samples, which showed weak expression of the PL-A/B genes. PRL- and GH-receptors were expressed in all normal and pathological prostatic specimens. Surprisingly, PRL-receptor expression was not detectable in prostatic cancer cell lines. The trophic effects of exogenous hGH, hPL, and hPRL were investigated by cell proliferation assays (WST-I) in prostatic primary cell cultures and PCa cell lines. hGH significantly (p < 0.005) increased cell proliferation up to 138+/-3.2% (1 nM hGH), while hPL and hPRL revealed only moderate effects. Our data suggest that local auto/paracrine networks of protein hormone actions are not involved in the pathology of BPH or prostatic cancer. On the other hand, systemic pituitary-derived hGH can increase the proliferative response of BPH and PCa, acting directly on the target organ prostate, via the hGH-R. In this case, envisaged GH substitution in elderly people must be viewed at with caution because age-related declines in GH/IGF-I could act as a protective mechanism against abnormal cell growth. PMID- 10363794 TI - Paradoxical increase of LDL-R expression on the surface of lymphocytes from healthy old (>65a) SENIEUR protocol-compatible donors compared to healthy young (<35a) controls. PMID- 10363795 TI - Experiments on a new phosphine-peptide chelator for labelling of peptides with Tc 99m. AB - A phosphine-containing ligand providing a N-[N-[3 (diphenylphosphino)propionyl]glycyl]-L-S-benzyl-cystein (PNNS) donor atomset for the chelation of 99mTc was studied in labelling experiments with a model peptide (tetragastrin, cholecystokinin-fragment). The peptide was conjugated to the ligand chelator by active ester chemistry either before or after radiolabelling. Both the chelator-conjugate and the preformed chelate approaches resulted in the same radiolabelled isomers of the ligand peptide. Sequence and reaction conditions influence yield and purity. PMID- 10363796 TI - Preclinical evaluation of 4-[18F]fluoroprolines: diastereomeric effect on metabolism and uptake in mice. AB - The aim of this study was to evaluate the diastereomeric effect on uptake and metabolic behavior of (2S,4R)-4-[18F]fluoro-L-proline (trans-[18F]FPro) and (2S,4S)-4-[18F]fluoro-L-proline (cis-[18F]FPro) in view of their potential suitability as tracers for abnormal collagen synthesis. No-carrier-added 4 [18F]fluoro-L-prolines were prepared according to the literature in about 150 min (50-60% radiochemical yield). Both compounds exhibited high in vivo stability. The tumor uptake of cis-[18F]FPro in osteosarcomas of mice was high and at 240 min postinjection reached 11.8 +/- 2.2 %ID/g compared with 7.07 +/- 1.68 %ID/g for trans-[18F]FPro. In contrat to trans-[18F]FPro, which showed fast and complete renal clearance, the cis isomer was accumulated in the pancreas, and showed hepatic clearance and renal reuptake. Speciation studies on tissue homogenates revealed protein incorporation only for cis-[18F]FPro. However, due to the relatively slow protein incorporation rate of cis-[18F]FPro, the tumor uptake of both compounds in colon carcinomas, mammary carcinomas, and osteosarcomas 1 h postinjection predominantly reflect amino acid transport. PMID- 10363797 TI - In vitro and in vivo evaluation of 64Cu-TETA-Tyr3-octreotate. A new somatostatin analog with improved target tissue uptake. AB - Radiolabeled somatostatin analogs have demonstrated potential as cancer therapeutic agents. Many of these agents are based on the analog octreotide (OC). Recently it has been shown that substitution of a tyrosine for phenylalanine in the 3-position and changing the C-terminus from an alcohol to an acid improves the targeting of somatostatin-rich tissues. The compound, 1,4,8,11 tetraazacyclotetradecane-N,N',N",N"'-tetraacetic acid-Tyr3-octreotate (TETA-Y3 TATE), was synthesized and radiolabeled with 64Cu. The receptor binding properties of 64Cu-TETA-Y3-TATE showed an estimated Kd value of 549 pM in somatostatin receptor-positive CA20948 tissue membrane. High tumor uptake was observed in two animal tumor models. Tumor uptakes of 2.37 %ID/g in CA20948 tumor bearing rats and 21.60 %ID/g in AR42J tumor-bearing SCID mice were observed at 1 h, compared with 1.09 %ID/g and 11.24 %ID/g for 64Cu-TETA-OC. Higher uptake in other somatostatin-receptor rich tissues was also observed, compared with 64Cu TETA-OC. Positron emission tomography (PET) imaging with 64Cu-TETA-Y3-TATE in a baboon showed significant uptake in the pituitary and adrenals, and clearance through the kidneys. 64Cu-TETA-Y3-TATE, a new OC analog for binding somatostatin receptors, demonstrated significantly greater uptake in somatostatin-rich tissues in two tumor-bearing animal models, and demonstrated great potential as a radiopharmaceutical for imaging and therapy of somatostatin receptor-positive tissues. PMID- 10363798 TI - Biodistribution of 99mTc-labeled anti-human epidermal growth factor receptor (EGF R) humanized monoclonal antibody h-R3 in a xenograft model of human lung adenocarcinoma. AB - The anti-human epidermal growth factor receptor (EGF-R) humanized monoclonal antibody (MAb) h-R3 is an (IgG1), which binds to an extracellular domain of EGF R. It was used to evaluate the biodistribution on nude mice xenografted with H 125 human lung adenocarcinoma cell line. Results were compared with its murine version of the MAb ior-egf/r3. Twenty-one athymic female 4NMRI nu/nu mice were injected intraperitoneally with 10 microg/100 muCi of 99mTc-labeled MAbs. Immunoreactivity of 99mTc-labeled MAbs were measured by enzyme-linked immunosorbent assay (ELISA) on H-125 cell line and the immunoreactive fractions was determined by the Lindmo method. Among all organs, significant accumulation was found in serum (27.05 +/- 2.08 %ID/g) and tumor (3.903 +/- 0.89 %ID/g) at 4 h after injection. These values decreased to 5.03 +/- 0.50 %ID/g and 2.19 +/- 0.56 %ID/g for serum and tumor, respectively. The immunoreactive fraction was found to be 0.70, with a correlation coefficient r = 0.9984. With the good biodistribution and tumor uptake of the 99mTc-labeled humanized antibody h-R3, a phase I diagnostic clinical trial of tumor with epithelial origin should be pursued. PMID- 10363799 TI - Species difference in radioactivity elimination from liver parenchymal cells after injection of radiolabeled proteins. AB - To elucidate the cause for the different levels of hepatic radioactivity among mammals after injection of protein radiopharmaceuticals, the metabolism of radiolabeled proteins and the fate of their radiometabolites in the parenchymal cells of rat liver were investigated and compared with those of mice. We used galactosyl-neoglycoalbumin (NGA) as a carrier protein, and NGA was labeled with 111In via 1-(4-isothiocyanatobenzyl)ethylenediaminetetraacetic acid (SCN-Bz-EDTA) or 1-[p-(5-maleimidopentyl)aminobenzyl]ethylenediaminetetraacetic acid (EMCS-Bz EDTA) and with 125I via direct iodination. All radiolabeled NGAs exhibited rapid accumulation in liver parenchymal cells after intravenous injection into rats. Radioactivity was eliminated following NGA-125I injection at similar rates from rat and mouse liver. In contrast, both 111In-labeled NGAs demonstrated much slower elimination of radioactivity in rat when compared with mouse liver. Analyses of radioactivity in bile and liver indicated that both SCN-Bz-EDTA and EMCS-Bz-EDTA rendered mono-amino acid adducts as the final radiometabolites, which were generated in rat liver within 1 h postinjection. Subcellular distribution studies suggested that these radiometabolites were copurified with lysosome in rat liver. Because similar results were observed in mice previously, the difference between rats and mice in radioactivity elimination from liver parenchymal cells would be predominantly attributable to the different efflux rate of the 111In-labeled metabolites from the lysosome between these species. Such differences in the efflux rates of radiometabolites from the lysosome among mammals may also account for the different hepatic radioactivity levels of radiolabeled proteins between animal and clinical studies. PMID- 10363800 TI - Quantitative ex vivo and in vitro receptor autoradiography using 11C-labeled ligands and an imaging plate: a study with a dopamine D2-like receptor ligand [11C]nemonapride. AB - Ex vivo and in vitro autoradiography (ARG) with radioluminography is a useful technique to characterize newly developed 11C-labeled positron emission tomography (PET) tracers and to apply them to biological and pharmacological studies. In this report, we have described a method of evaluating the radioactivity distribution quantitatively in ex vivo and in vitro ARG using imaging plates and a dopamine D2-like receptor ligand [11C]nemonapride as a model compound. The photo-stimulated luminescence (PSL) values of the rat brain section provided by the imaging plates showed an excellent linear relationship with the radioactivity in a wide range under constant slice-thickness, although the PSL values slightly decreased with increasing slice-thickness both in ex vivo and in vitro ARG. The injection dose of 11C-tracers for ex vivo ARG was also discussed. We found saturable binding sites of [11C]nemonapride in the cortex besides the striatum both ex vivo and in vitro. PMID- 10363801 TI - Characterization and preliminary evaluation of ester-modified technetium-99m SNS/S mixed ligand complexes as potential brain perfusion agents. AB - Two novel [99mTc](SNS/S) mixed ligand complexes carrying a pendant ester function on the monothiolate coligand were synthesized. The corresponding oxorhenium and [99gTc]oxotechnetium complexes prepared at the macroscopic level and chemically characterized were used for structure assignment of [99mTc](SNS/S) complexes prepared at the nanomolar level. Enzymatic hydrolysis of the pendant ester group of [99mTc](SNS/S) mixed ligand complexes by esterase was investigated in vitro and compared with that of the ethyl cysteinate dimer, [99mTc]ECD. Preliminary biodistribution data in mice shows that the complexes are lipophilic and exhibit significant initial uptake in rodent brain. PMID- 10363802 TI - Evaluation of gallium-68 tris(2-mercaptobenzyl)amine: a complex with brain and myocardial uptake. AB - Previous research into development of a gallium-radiolabeled agent that crosses the blood-brain barrier has met with limited success. In this study, we focused our attention on a Ga(III) complex of a 4-coordinate amine trithiolate tripod ligand, tris(2-mercaptobenzyl) amine (S3N). The Ga(III) S3N complex is small, neutral, and lipophilic, meeting the requirements for a potential brain imaging agent. The Ga-68 complex was easily formed with a radiochemical purity of >95%. In vitro stability of the Ga-S3N complex, determined in rat serum incubated at 37 degrees C, was greater than 95% intact at 2 h by silica gel and reversed-phase radio-thin layer chromatography. Biodistribution studies conducted in female Sprague-Dawley rats showed the complex cleared rapidly from the blood with initial high liver uptake followed by rapid washout. Significant uptake was observed in the brain, with brain:blood ratios increasing from 0.11 at 2 min postinjection to 3.8 at 60 min postinjection. Uptake was also observed in the heart going from a heart:blood ratio of 2.3 at 2 min postinjection to 11 at 60 min postinjection. Molecular mechanics were used to determine the coordination number, and demonstrated that the Ga(III) complex prefers to be 4-coordinate. Imaging studies with 68Ga-S3N in a Nemestrina macaque showed significant brain uptake, similar to other lipophilic agents. The extraction of 68Ga-S3N into the brains of both rodents and primates, higher than any 68Ga agent reported in the literature, suggests that this compound may have potential as a brain imaging agent for positron emission tomography. PMID- 10363803 TI - New preparation of (123I) iodolisuride from the 2-tri-n-butylstannalisuride derivative. PMID- 10363804 TI - The pharmacokinetics of [11C]methoxy-norchloroprogabidic acid, a potential PET tracer for GABA receptors in the brain. PMID- 10363806 TI - Evaluation of methods for large scale preparation of antibody ligand conjugates. PMID- 10363805 TI - On the synthesis, isolation, and radiochemical studies for the preparation of in house kits for 99mTc-meso- and d,l-HMPAO: a few additional observations. PMID- 10363807 TI - Gamma-band oscillations in the "barrel cortex" precede rat's exploratory whisking. AB - Cortical oscillations have been reported in relation to animal behavior and may have important roles in neuronal information processing such as "binding". We considered oscillatory activities in the primary somatosensory cortex of rats as a preparatory state for sensory processing and investigated the temporal relationship between gamma oscillation (25-45 Hz) onset in the somatosensory cortex (the barrel cortex) and rat's whisking as tactile exploratory behavior. The gamma oscillations of local field potentials preceded the whisking in 52/83 cases (63%). The mean lead time of the oscillations, defined as time from the oscillation onset to the whisking, was 268 ms (+/- 123, S.D.). These preceding gamma oscillations lasted to the onset of whisking. Furthermore, we investigated the temporal relationship during the other two behavioral states: a still but alert state and a state with tactile stimulation of the whiskers. In the state with tactile stimulation, gamma oscillations followed the stimulation, and in the still but alert state, significant gamma oscillation was not observed. The gamma oscillations that precede the whisking are thought to reflect anticipatory activities in the barrel cortex for the subsequent somatosensory input from the whiskers, which could represent a mechanism for rapid processing of expected sensory information. PMID- 10363808 TI - Lateral evaginations from the third ventricle into the rat mediobasal hypothalamus: an amplification of the ventricular route. AB - In this work we report the existence of several evaginations extending out of the third ventricle within the mediobasal hypothalamus of the rat. In coronal sections, these evaginations appear as very narrow canaliculi integrating a canalicular system, which increases the contact surface between the ventricular lining and the nervous tissue. Consequently these evaginations enlarge the ventricular route for the transport of active principles present in the cerebrospinal fluid, such as (neuro)hormones and neurotransmitters. The mediobasal hypothalamus includes the arcuate nucleus and the median eminence (both involved in neuroendocrine mechanisms and in the regulation of pituitary function). A possible implication of our finding is that the neuroactive substance-containing ventricular cerebrospinal fluid may reach the intercellular spaces of the surrounding neuropil of the arcuate nucleus. According to literature these substances cross the ependyma of the lateral wall of the infundibular recess of the third ventricle. We suggest that such substances might also pass through the ependymal lining of the canalicular system, which displays the same ultrastructural characteristics as the rest of the ependyma of the lateral wall of the third ventricle. Therefore, the arcuate neurons may be influenced not only by synaptic inputs (afferent fibers) but also by non-synaptic diffusion neurotransmission (by means of neuroactive substances present in the cerebrospinal fluid). PMID- 10363809 TI - Endogenous neurotrophin-3 is retrogradely transported in the rat sciatic nerve. AB - To address the active transport of neurotrophins, nerve growth factor, brain derived neurotrophic factor, and neurotrophin-3 in the peripheral nerves, we examined the levels of proteins and messenger RNAs in the sciatic nerve of adult rats following transection, using enzyme immunoassays and reverse transcription polymerase chain reaction method, respectively. Neurotrophin-3 protein increased one day after transection only in the distal segment next to the transection site and returned to the original level two days later. This was considered to reflect accumulation of neurotrophin-3 transported from the periphery toward the neuronal cell bodies, because the neurotrophin-3 messenger RNA level was not changed in any sciatic segments during this experimental period. An increase in brain derived neurotrophic factor protein was observed simultaneously in both the distal and proximal stumps three days after transection. Brain-derived neurotrophic factor messenger RNA was elevated in the same stumps two days after transection, suggesting that brain-derived neurotrophic factor was produced within the transected stumps. These observations demonstrate that neurotrophin-3, like nerve growth factor, is retrogradely transported in the sciatic nerve but that brain-derived neurotrophic factor is not. This suggests that neurotrophin-3 plays a role in the conveyance of trophic signals from target organs to neurons. PMID- 10363810 TI - A novel role for receptor-associated protein in somatostatin modulation: implications for Alzheimer's disease. AB - Receptor-associated protein appears to play an important role in low-density lipoprotein receptor-related protein trafficking. Since ligands for the low density lipoprotein receptor-related protein have been implicated in Alzheimer's disease and normal functioning of this protein is indispensable for central nervous system development, deficient receptor-associated protein expression may result in central nervous system alterations. In this study, receptor-associated protein knockout mice were behaviorally tested and nervous system integrity was assessed via in situ hybridization and immunocytochemical/laser confocal microscopy methods. Receptor-associated protein knockout mice were found to be cognitively impaired in the Morris water maze compared to controls. In wild-type mice, the receptor-associated protein was found to be highly co-expressed with somatostatin in hippocampal and neocortical inhibitory neurons. Receptor associated protein knockout mice, however, showed a significant decrease in number of somatostatin-expressing neurons of the CA1 region and somatostatin expression within these neurons. The decreased number of somatostatin neurons significantly correlated with cognitive impairment observed in the receptor associated protein knockout mice. These results suggest a novel role for receptor associated protein in modulating the functioning of somatostatin-producing neurons. Furthermore, this has implications for Alzheimer's disease pathogenesis, in which altered regulation of both somatostatin and the known low-density lipoprotein receptor-related protein ligands are a consistent finding. PMID- 10363811 TI - The entorhino-septo-supramammillary nucleus connection in the rat: morphological basis of a feedback mechanism regulating hippocampal theta rhythm. AB - Recent electrophysiological observations suggest that, in addition to the medial septal area pacemaker system, several alternative or additional mechanisms are involved in the generation/regulation of hippocampal theta activity. Discharging neurons phase-locked to hippocampal theta waves have been observed in the dorsal raphe, nucleus reticularis pontis oralis and especially in the supramammillary region of rats. Since these areas are reciprocally interconnected with the hippocampal formation, including the entorhinal cortex, it would aid our understanding of limbic function to elucidate the location and neurochemical content of the entorhino-septal and septo-supramammillary projection neurons, as well as that of their postsynaptic targets. Light and electron microscopic immunostaining for calretinin, in combination with antero- and retrograde tracer techniques, postembedding immunostaining for GABA and the transmitter specific [3H]D-aspartate retrograde radiolabeling, as well as a co-localization experiment for calretinin and glutamate decarboxylase in rat supramammillary and septal neurons, demonstrated that: (i) a large population of entorhinal cells that forms asymmetric synaptic contacts on calretinin-containing neurons located at the border between the medial and lateral septal areas contains calretinin and are aspartate/glutamatergic; (ii) the overwhelming majority of calretinin immunoreactive cells located at the border between the lateral and medial septal area are GABAergic; (iii) these neurons can be retrogradely labeled from the supramammillary area; (iv) anterogradely labeled axons originating in the border between the medial and lateral septum are GABAergic and (v) terminate on supramammillary area non-GABAergic, calretinin-containing neurons, which are known to project to the septal complex and hippocampus. These observations indicate that a large population of cells participating in the hippocampal feedback regulation of theta regulation/generation contain the same calcium binding protein. Furthermore, entorhinal excitatory transmitter-containing neurons can depress the activity of supramammillary theta generating/regulating cells via septal inhibitory neurons. PMID- 10363812 TI - Differential time-course of slow afterhyperpolarizations and associated Ca2+ transients in rat CA1 pyramidal neurons: further dissociation by Ca2+ buffer. AB - Hippocampal neurons exhibit a slow afterhyperpolarization following membrane depolarization; this is thought to reflect an underlying Ca2+-dependent K+ current. This current is potentiated by intermediate concentrations (0.1-1.0 mM) of exogenous Ca2+ buffer [Schwindt P. C. et al. (1992) Neuroscience 47, 571-578; Zhang L. et al. (1995) J. Neurophysiol. 74, 2225-2241]. The relationship between the slow afterhyperpolarization and associated Ca2+ transients was investigated in the presence and absence of added exogenous Ca2+ buffer. Slow afterhyperpolarizations and underlying K+ currents were measured using whole-cell patch-clamp recordings from hippocampal CA1 neurons in acute rat brain slices. Inclusion of fluorescent Ca2+ indicators in the patch pipette solution allowed simultaneous measurement of the evoked subcellular Ca2+ transients using a confocal microscope. The peak Ca2+ signal exhibited an incremental increase with each action potential. This increase eventually reached a plateau with increasing numbers of action potentials, suggesting dye saturation with peak Ca2+ concentrations. As the K(D) for Ca2+ of the indicator dyes used was between 200 and 300 nM, it is predicted that saturation will occur when the peak Ca2+ signal exceeds 1 microM. This occurred with fewer action potentials in dendritic vs somatic compartments. Neither compartment exhibited averaged Ca2+ transients matching the slow afterhyperpolarization time-course, dendritic Ca2+ transients being most divergent. Intracellular accumulation of exogenous Ca2+ buffer, either by inclusion in the patch pipette or by incubation of the brain slice with its membrane-permeable form, caused a prolongation of the slow afterhyperpolarization but not of the somatic Ca2+ transient. The initial rate of decline of the dendritic Ca2+ transient was diminished, but remained faster than that of the slow afterhyperpolarization. We conclude that neither dendritic nor somatic Ca2+ signals match the slow afterhyperpolarization time-course, with this dissociation being further magnified by addition of exogenous Ca2+ buffer. The implications of this result are discussed. PMID- 10363813 TI - Reduced Mg2+ blockade of synaptically activated N-methyl-D-aspartate receptor channels in CA1 pyramidal neurons in kainic acid-lesioned rat hippocampus. AB - Unilateral kainic acid lesion in the hippocampus caused a long-term change in the balance between excitatory and inhibitory drive onto CA1 pyramidal cells, making these cells hyperexcitable several weeks post-lesion. In this study, we have shown an enhanced N-methyl-D-aspartate receptor-mediated component in the excitatory synaptic transmission together with a reduced GABA(A) receptor mediated inhibition in CA1 pyramidal cells one-week post kainic acid lesion. In these cells, pharmacologically isolated N-methyl-D-aspartate receptor-mediated whole-cell excitatory postsynaptic currents were significantly larger at negative holding potentials, and the voltage-dependence of N-methyl-D-aspartate receptor channels was shifted in the hyperpolarizing direction. The plot of relative conductance (g/gMax) shifted significantly (P<0.01) to more negative holding potentials by 19 mV (-28+/-4 mV in control slices and -47+/-4 mV in kainic acid slices) at the half maximal conductance point (g/gMax =0.5). This shift gives a larger N-methyl-D-aspartate receptor-mediated component in the excitatory synaptic transmission at resting membrane potentials (around -60 mV). The shifted voltage dependence is highly sensitive to extracellular Mg2+ ions. Moderate increases in [Mg2+]o from 1 mM to 2.6 mM more than compensated for the negative shift and effectively suppressed the population epileptiform bursting activity. Fitting the voltage dependence to an ionic block model revealed a higher dissociation constant of N-methyl-D-aspartate receptor channels for Mg2+ in kainic acid-lesioned slices (52 mM at 0 mV; 330 microM at -60 mV) than in control slices (7.7 mM at 0 mV; 93 microM at -60 mV). While a simple single site model adequately fitted the control data for [Mg2+]o at 1 mM and 2.6 mM, no consistent model of this form was found for the kainic acid-lesioned slices. These results revealed changed properties of N-methyl-D-aspartate receptor channels in the kainic acid-lesioned model of epilepsy. The reduced Mg2+ blockade of N-methyl-D aspartate receptor channels contributed significantly to the epileptiform bursting activity. PMID- 10363814 TI - Restoration of decaying long-term potentiation in the hippocampal formation by stimulation of neuromodulatory nuclei in freely moving rats. AB - Induction of long-term potentiation within the hippocampal formation can be modulated by afferent influences from a number of subcortical structures known to be involved in hippocampal-dependent learning and memory. This study performed on freely moving rats investigated the effects of stimulation of the noradrenergic locus coeruleus nucleus and the serotonergic dorsal raphe nucleus on spontaneously decaying posttetanic long-term potentiation in the dentate gyrus and the hippocampal CA1 area, respectively. High-frequency electrical stimulation of the locus coeruleus or the dorsal raphe elicited a well-expressed behavioural reaction of exploratory or defensive type, respectively, but did not significantly alter transmission at perforant path-dentate gyrus or Schaffer collateral-CA synapses, when delivered either before tetanic stimulation of the perforant path or the Schaffer collaterals or long (hours and days) after previously induced long-term potentiation had completely decayed. However, when locus coeruleus or dorsal raphe stimulation was delivered with the same parameters during a limited time (minutes and hours) after marked or even complete decay of tetanus-induced long-term potentiation at perforant path dentate gyrus or Schaffer collateral-CA1 synapses, the potentiation was partially or entirely restored but never increased beyond the initial level of potentiation. In CA1, stimulation of ipsilateral and contralateral Schaffer collaterals demonstrated that the restoration of previously existing long-term potentiation by dorsal raphe stimulation was input-specific, occurring, like tetanus-induced potentiation, only in the pathway which had previously been tetanized. These findings suggest that the noradrenergic locus coeruleus and the serotonergic dorsal raphe can influence not only induction, but also spontaneous decay of long-term potentiation in the hippocampal formation. Since hippocampal long-term potentiation is thought to play a role in certain kinds of learning and memory, and association of tetanic stimulation with activation of ascending neuromodulatory systems is required for full expression of long-term potentiation, the restoration of hippocampal long-term potentiation by activation of a neuromodulatory system alone may serve as a mechanism of associative reminder which may underlie facilitation of memory retrieval after a period of forgetting, as has been observed in trained rats under similar conditions. PMID- 10363815 TI - Distributions of nicotinic acetylcholine receptor alpha7 and beta2 subunits on cultured hippocampal neurons. AB - The hippocampus receives cholinergic afferents and expresses neuronal nicotinic acetylcholine receptors. In particular, the alpha7 and beta2 nicotinic subunits are highly expressed in the hippocampus. There has been controversy about the location, distribution and roles of neuronal nicotinic acetylcholine receptors [Role L. W. and Berg D. K. (1996) Neuron 16, 1077-1085; Wonnacott S. (1997) Trends Neurosci. 20, 92-98]. Using immunocytochemistry and patch-clamp techniques, we examined the density and distribution of nicotinic receptors on rat hippocampal neurons in primary tissue culture. The density and distribution of alpha7 subunits change with days in culture. Before 10 days in culture, alpha7 expression, monitored immunocytochemically, is low and nicotinic currents are small or absent. In older cultures, about two-thirds of the neurons express nicotinic currents, and alpha7 appears in small patches on the soma and out along the neuronal processes. These patches of alpha7 subunits on the surface of the neuronal processes often co-localize with the presynaptic marker, synaptotagmin. The other most common nicotinic subunit, beta2, stays confined mainly to the soma and proximal processes, and beta2 is distributed more uniformly and is not specifically localized at presynaptic areas. The two subunits, alpha7 and beta2, have different expression patterns on the surface of the cultured hippocampal neurons. Taken together with previous physiological studies, the results indicate that alpha7 subunits can be found at presynaptic terminals, and at these locations, these calcium-permeable channels may influence transmitter release. PMID- 10363816 TI - Expression of cocaine sensitization: regulation by the medial prefrontal cortex. AB - Extracellular levels of dopamine are increased in response to systemic administration of cocaine in several brain areas including the nucleus accumbens and medial prefrontal cortex. While the cocaine-induced increase in extracellular dopamine levels in the nucleus accumbens is augmented after repeated daily cocaine, the response of extracellular dopamine levels in the medial prefrontal cortex is attenuated. Since dopamine in the medial prefrontal cortex has an inhibitory effect on nucleus accumbens dopamine levels and locomotor activity, the role of medial prefrontal cortex dopamine tolerance in the expression of sensitized locomotor behavior was further examined by injection of D-amphetamine sulfate into the prelimbic portion of the medial prefrontal cortex just prior to cocaine challenge in cocaine-sensitized rats. Male Sprague-Dawley rats were non handled (naive) or injected with either saline (1 ml/kg, i.p.) or cocaine (15 mg/kg, i.p.) for five consecutive days. After a seven to 12 day withdrawal period, rats were microinjected with either saline or various doses of amphetamine into primarily the prelimbic region of the medial prefrontal cortex followed by systemic injection of saline or cocaine. In naive rats, intramedial prefrontal cortex amphetamine produced a trend toward decreased locomotor responding to cocaine challenge while no effect of amphetamine was evident in daily saline pretreated rats. Daily cocaine pretreated rats that received saline in the medial prefrontal cortex demonstrated a sensitized locomotor response compared to their daily saline pretreated counterparts. This sensitization was blocked by a low dose of amphetamine (0.175 microg/side) in the medial prefrontal cortex, an effect which disappeared in animals administered higher amphetamine doses. The results suggest that in rats sensitized to cocaine, decreased medial prefrontal cortex dopamine levels in response to cocaine challenge may contribute to behavioral sensitization. Furthermore, the data indicate the possibility that there is an optimal range at which medial prefrontal cortex amphetamine exerts maximal behavioral inhibition. These findings implicate a role for decreased cortical control in producing sensitized behavioral responding to cocaine. PMID- 10363817 TI - The distribution of dynorphinergic terminals in striatal target regions in comparison to the distribution of substance P-containing and enkephalinergic terminals in monkeys and humans. AB - Single- and double-label immunohistochemical techniques using several different highly specific antisera against dynorphin peptides were used to examine the distribution of dynorphinergic terminals in globus pallidus and substantia nigra in rhesus monkeys and humans in comparison to substance P-containing and enkephalinergic terminals in these same regions. Similar results were observed in monkey and human tissue. Dynorphinergic fibers were very abundant in the medial half of the internal pallidal segment, but scarce in the external pallidal segment and the lateral half of the internal pallidal segment. In substantia nigra, dynorphinergic fibers were present in both the pars compacta and reticulata. Labeling of adjacent sections for enkephalin or substance P showed that the dynorphinergic terminals overlapped those for substance P in the medial half of the internal pallidal segment, but showed only slight overlap with enkephalinergic terminals in the external pallidal segment. The substance P containing fibers were moderately abundant along the borders of the external pallidal segment, and enkephalinergic fibers were moderately abundant in parts of the internal pallidal segment. Dynorphinergic and substance P-containing terminals overlapped extensively in the nigra, and both extensively overlapped enkephalinergic fibers in medial nigra. Immunofluorescence double-labeling studies revealed that dynorphin co-localized extensively with substance P in individual fibers and terminals in the medial half of the internal pallidal segment and in substantia nigra. Thus, as has been found in non-primates, dynorphin within the striatum and its projection systems appears to be extensively localized to substance P-containing striatopallidal and striatonigral projection neurons. Nonetheless, our results also raise the possibility that a population of substance P-containing neurons that projects to the internal pallidal segment and does not contain dynorphin is present in primate striatum. Our results also suggest the possible existence of populations of striatopallidal and striatonigral projection neurons in which substance P and enkephalin or dynorphin and enkephalin, or all three, are co-localized. Thus, striatal projection neurons in primates may not consist of merely two types, one containing substance P and dynorphin and the other enkephalin. PMID- 10363819 TI - Role of accumbens and cortical dopamine receptors in the regulation of cortical acetylcholine release. AB - Cortical acetylcholine, under resting and stimulated conditions, was measured in frontoparietal and prefrontal cortex using in vivo microdialysis in freely-moving rats. Cortical acetylcholine efflux was stimulated by systemic administration of the benzodiazepine receptor partial inverse agonist FG 7142. Administration of FG 7142 (8.0 mg/kg; i.p.) significantly elevated acetylcholine efflux in both cortical regions (150-250% relative to baseline) for 30 min after drug administration. The ability of endogenous dopamine to regulate cortical acetylcholine efflux under resting or stimulated conditions and the relative contributions of D1- and D2-like dopamine receptor activation was also assessed. In a first series of experiments, systemic administration of the antipsychotic drug haloperidol (0.15, 0.9 mg/kg, i.p.) blocked FG 7142-stimulated acetylcholine efflux in frontoparietal, cortex while the D1-like antagonist, SCH 23390 (0.1, 0.3 mg/kg), was less effective in attenuating stimulated acetylcholine efflux. In a second series of experiments, the effects of infusions of these antagonists and of the D2-like antagonist sulpiride (10, 100 microM) into the nucleus accumbens were assessed. Infusions of haloperidol and sulpiride significantly blocked FG 7142-stimulated acetylcholine efflux while SCH 23390 did not. By contrast, a third series of experiments demonstrated that perfusion of these antagonists (100 microM) locally into the cortex (through the probe) did not affect FG 7142 stimulated acetylcholine efflux. Moreover, none of these dopamine receptor antagonists, whether administered systemically or perfused into the nucleus accumbens or cortex, affected basal cortical acetylcholine efflux. These results reveal similarities in stimulated cortical acetylcholine release across frontal cortical regions and suggest a prominent role for D2-mediated accumbens dopamine transmission in the regulation of cortical acetylcholine release. The findings provide evidence in support of a neural substrate that links dysregulation of mesolimbic dopaminergic transmission to changes in cortical cholinergic transmission. Dysregulation within this circuit is hypothesized to contribute to the etiology of disorders such as schizophrenia, dementia and drug abuse. PMID- 10363818 TI - Enkephalin regulates acute D2 dopamine receptor antagonist-induced immediate early gene expression in striatal neurons. AB - Projection neurons of the striatum release opioid peptides in addition to GABA. Our previous studies showed that the opioid peptide dynorphin regulates that subtype of projection neurons which sends axons to the substantia nigra/entopeduncular nucleus, as indicated by an inhibitory action of dynorphin/agonists on D1 dopamine receptor-mediated immediate-early gene induction in these neurons. The other subtype of striatal projection neurons projects to the globus pallidus and contains the opioid peptide enkephalin. Here, we investigated whether enkephalin regulates the function of striatopallidal neurons, by analysing opioid effects on immediate-early gene induction by D2 dopamine receptor blockade that occurs in these neurons. Thus, the effects of systemic and intrastriatal administration of various opioid receptor agonists and antagonists on immediate-early gene expression (c-fos, zif 268) induced by the D2 receptor antagonist eticlopride were examined with in situ hybridization histochemistry. Intrastriatal infusion of enkephalin (delta and mu), but not dynorphin (kappa), receptor agonists suppressed immediate-early gene induction by eticlopride in a dose-dependent manner. This suppression was blocked by the opioid receptor antagonist naloxone, confirming the involvement of opioid receptors. Repeated treatment with D2 receptor antagonists produces increased enkephalin expression and diminished immediate-early gene inducibility in striatopallidal neurons, as well as behavioral effects that are attenuated compared to those of acute treatment (e.g., reduced akinesia). Naloxone reversed such behavioral recovery (i.e. reinstated akinesia), but did not significantly affect suppressed immediate-early gene induction. Our results indicate that enkephalin acts, via mu and delta receptors in the striatum, to inhibit acute effects of D2 receptor blockade in striatopallidal neurons. Moreover, the present findings suggest that increased enkephalin expression after repeated D2 receptor antagonist treatment is an adaptive response that counteracts functional consequences of D2 receptor blockade, but is not involved in suppressed immediate early gene induction. Together with our earlier findings of the role of dynorphin, these results indicate that opioid peptides in the striatum serve as negative feedback systems to regulate the striatal output pathways in which they are expressed. PMID- 10363820 TI - Variations of nucleus accumbens dopamine and serotonin following systemic interleukin-1, interleukin-2 or interleukin-6 treatment. AB - The effects of systemically administered interleukin-1beta (1.0 microg), interleukin-6 (1.0 microg) and interleukin-2 (1.0 microg) on in vivo variations of monoamines were assessed in the nucleus accumbens. Administration of interleukin-1beta did not affect extracellular accumbal dopamine, provoked a modest rise of homovanillic acid, and prevented the decline of dihydroxyphenylacetic acid ordinarily seen in saline treated rats. Also, interleukin-1 provoked a modest increase of extracellular 5-hydroxyindoleacetic acid from the nucleus accumbens. Following exposure to the stress of a series of air-puffs, a still greater increase of accumbal 5-hydroxyindoleacetic acid was evident. In contrast to interleukin-1, systemic administration of interleukin-6 and interleukin-2 both induced marked reductions of interstitial dopamine levels. The air-puff exposure further enhanced these effects in rats that had received the cytokine treatment. As well, interleukin-6 and interleukin-2 were both found to reduce the homovanillic acid response associated with the stress, and interleukin-2 promoted a decline of homovanillic acid levels. Treatment with interleukin-6, like that of interleukin-1, prevented the decline of dihydroxyphenylacetic acid ordinarily observed over time, while interleukin-2 was without effect in this respect. Finally, interleukin-6 provoked a modest rise of 5-hydroxyindoleacetic acid, which was most apparent following air-puff exposure, while administration of interleukin-2 did not affect accumbal 5 hydroxyindoleacetic acid. It is suggested that the cytokines may influence the release of biogenic amines in the nucleus accumbens, but the profile of changes were cytokine-specific. As well, it appeared that the cytokines, particularly interleukin-1 and interleukin-6, may act synergistically with the stressor in promoting the amine variations. Systemic administration of cytokines clearly influenced monoamine activity at the nucleus accumbens, a region associated with both rewarding and aversive events. Thus, it may be expected that cytokine treatments may affect behavior. Moreover, it seems that the effects of interleukin-1 and interleukin-6 may be influenced by the presence of stressful stimuli. It ought to be underscored that although cytokines share features with the effects of stressors, most notably the variations of hypothalamic-pituitary adrenal hormones, the pattern of central neurochemical changes elicited by the cytokines could be distinguished from the amine variations ordinarily associated with stressors. PMID- 10363821 TI - Effects of corticosterone on excitatory amino acid responses in dopamine sensitive neurons in the ventral tegmental area. AB - The ventral tegmental area is involved in reward processes and in drug dependence and sends dopaminergic projections to the nucleus accumbens and prefrontal cortex. Stress, and glucocorticoid hormones, are thought to play an important role in the development of drug dependence, but there has been little investigation of the effects of these hormones on ventral tegmental function. The present study examined the effects of corticosterone on single-unit recordings from dopamine-sensitive neurons in the ventral tegmental area in midbrain slices. At concentrations of 100 nM and above, corticosterone potentiated the responses to N-methyl-D-aspartate. This effect was not seen when the calcium concentration of the bathing medium was reduced to 0.1 mM. Responses to alpha-amino-3-hydroxy-5 methylisoxazole-4-propionate (AMPA) and kainic acid were also considerably potentiated, at concentrations of corticosterone over 100 nM, while there was some evidence of decreases in these responses at the 100 nM concentration of this hormone. Aldosterone, at concentrations of 100 nM and above reduced the responses to N-methyl-D-aspartate, but had no effects at lower concentrations. RU38486, which acts as an antagonist at glucocorticoid (Type II) receptors, prevented the effects of corticosterone on responses to N-methyl-D-aspartate, with no effect on the spontaneous firing rate or on the effects of N-methyl-D-aspartate in the absence of corticosterone. The latter result, and the effects of aldosterone, suggest that the potentiation of responses to N-methyl-D-aspartate was mediated through Type II glucocorticoid receptors. This study suggests that potentiation of responses to excitatory amino acids by corticosterone may alter the function of ventral tegmental neurons during stress, and it is possible that this effect is involved in the development of drug dependence. PMID- 10363822 TI - Spatiotemporal expression gradients of the carbohydrate antigen (CD15) (Lewis X) during development of the human basal ganglia. AB - The developmental expression pattern of the carbohydrate epitope CD15 (Lewis X, Le X) (alpha1-->3-fucosyl-N-acetyl-lactosamine) has been immunocytochemically evaluated in paraffin sections within the human basal ganglia from 10 weeks gestation to three years after birth. At 11 weeks of gestation, CD15 (Le X) positive radial glial cells were located in the anterior and dorsal parts of the lateral ganglionic eminence. Their processes ran from the subventricular zone radially in a highly ordered fashion to the dorsolateral margin of the caudate nucleus and further to the lateral rim of the putamen. At 12 weeks of gestation, strands of CD15 (Le X) material continued to the pial surface, forming a continuous CD15 (Le X) positive borderline separating the accumbens nucleus and olfactory tubercle from the piriform cortex. At 13 weeks of gestation the dorsal putamen was completely CD15 (Le X) immunoreactive along its perimeter and CD15 (Le X) patches, consisting of fine granular material, appeared at the dorsolateral margin of the putamen at this age; while the first CD15 (Le X) patches in the caudate nucleus were observed four weeks later. The matrix compartment of the caudate and dorsal putamen became gradually stained by granular CD15 (Le X) positive material into which CD15 (Le X) immunoreactive somata were embedded. The striking contrast in staining between patch and matrix compartments disappeared shortly after birth. The ventral striatum did not become immunoreactive until the last few weeks before birth. After the formation of CD15 (Le X) positive patches in the striatum (from 12 weeks of gestation), delicate CD15 (Le X) fibres, often accumulated in bundles and related to the striatal patches, became apparent coursing towards the external pallidal lamina and the globus pallidus. Immunoreactivity in the globus pallidus itself was transient, emerging from 16 weeks of gestation, reaching a peak at 21 weeks of gestation and disappearing by birth. Both processes, i.e. the occurrence of CD15 (Le X) striatopallidal fibres and the emerging immunoreactivity in their pallidal target, may be interrelated, so that ingrowing CD15 (Le X) positive axons from the striatum provoke CD15 (Le X) expression in the external and internal pallidum. The variable patterns and intensities of CD15 (Le X) expression are possibly related to periods of maturation of the striatum and the establishment of functional interactions within the basal ganglia. Differential staining of patch and matrix in the developing neostriatum suggests that a distinct phase of cellular adhesion or dishesion mediated by the CD15 (Le X) epitope occurs during establishment of the patch and matrix regions. PMID- 10363823 TI - Vasoactive intestinal peptide neurons as synaptic targets for vasopressin neurons in the suprachiasmatic nucleus. Double-label immunocytochemical demonstration in the rat. AB - Cellular relationships between neurons producing vasopressin or vasoactive intestinal peptide in the suprachiasmatic nucleus of the hypothalamus, the main component of the central circadian timing system in mammals, were investigated in the rat using double immunocytochemistry. Analysis of serial confocal images revealed that the vasopressin-synthesizing neurons not only are important targets for the vasoactive intestinal peptide-synthesizing neurons, as previously demonstrated, but also establish reciprocal axosomatic contacts with these neurons, which have never been reported. On average, 5.4 vasoactive intestinal peptide contacts per vasopressin perikaryon and 1.7 vasopressin contacts per vasoactive intestinal peptide perikaryon were counted. That both types of neurons are linked by reciprocal synapses was confirmed at the electron microscopic level using a combination of immunoperoxidase and immunogold-silver labeling. Existence of an anatomical substrate for a vasopressinergic control of the vasoactive intestinal peptide neurons may have important functional consequences. In view (i) of the presumed, direct or indirect, involvement of the vasopressin neurons in relaying pacemaker information within and outside the suprachiasmatic nucleus, and (ii) of the established role of the vasoactive intestinal peptide neurons as the main light-sensitive cells, it provides support for a neuronal mechanism through which the circadian clock may regulate inputs related to environmental messages. Our electron-microscopic data also extended earlier observations, pointing to the involvement of vasopressin and vasoactive intestinal peptide terminals in so-called double synapses that, conceivably, could regulate neuronal synchronization in the suprachiasmatic nucleus. A morphological basis for non synaptic interactions that could be involved in ephaptic and/or paracrine communication between both types of peptidergic neurons is also reported. PMID- 10363824 TI - Monoaminergic long-term facilitation of GABA-mediated inhibitory transmission at cerebellar synapses. AB - Long-term facilitation of neurotransmission by monoaminergic systems is implicated in the cellular mechanism of memory and learning-related processes at invertebrate synapses. Using whole-cell recording and rat cerebellar slices, we have examined whether mammalian monoamine-containing neurons play analogous roles in synaptic plasticity, and our results suggest that serotonin and noradrenaline are critically involved in short- and long-term modulation of GABAergic transmission in the cerebellar cortex. Exogenously applied serotonin and noradrenaline selectively induced a short-term enhancement of GABAergic transmission between cerebellar interneurons and Purkinje cells, their effect subsiding in 30 min. Successive amine applications converted this effect to long term facilitation lasting more than 2 h. During the monoamine-induced short- and long-term facilitation, spontaneously occurring miniature inhibitory synaptic responses increased in frequency, without significant changes in their mean amplitude and amplitude distribution, as well as the GABA receptor sensitivity of Purkinje cells. The actions of the two amines on the inhibitory transmission were mimicked by forskolin and blocked by kinase inhibitors, H-7, H-89 and Rp adenosine 3',5'-cyclic monophosphothioate. Thus, serotonin and noradrenaline are likely to activate cyclic-AMP- and protein kinase-dependent pathways in GABAergic interneurons, thereby reinforcing the inhibitory transmission on to Purkinje cells. Repetitive electrical stimulation within the molecular layer mimicked the facilitatory effect induced by exogenous monoamines: namely, neural stimulation selectively elicited long-lasting enhancement of GABAergic transmission in a manner sensitive to the monoamine receptor antagonists, methiothepin and propranolol, and an uptake inhibitor, imipramine. Synaptically released monoamines thus appear to induce cyclic-AMP- and protein kinase-dependent long term facilitation of cerebellar GABAergic transmission, thereby providing a likely mechanism of synaptic plasticity associated with motor coordination within the mammalian cerebellar system. PMID- 10363825 TI - 5-Hydroxytryptamine2A receptor stimulation induces activator protein-1 and cyclic AMP-responsive element binding with cyclic AMP-responsive element-binding protein and Jun D as common components in cerebellar neurons. AB - Previous studies from our laboratory have demonstrated that stimulation of 5 hydroxytryptamine2A receptors in rat cerebellar granule cells produces an increase in the levels of 5-hydroxytryptamine2A receptor messenger RNA and binding sites, and that this up-regulation requires de novo RNA and protein synthesis. Here we showed that up-regulation of 5-hydroxytryptamine2A receptor binding sites induced by stimulation with the 5-hydroxytryptamine2A/2C receptor agonist, (+/-)-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI), is associated with an increase in the 5-hydroxytryptamine2A receptor transcription rate. To examine the possible role of transcriptional activation in DOI-induced 5 hydroxytryptamine2A receptor up-regulation, we studied the effects of DOI on transcription factor binding to activator protein-1 and cyclic AMP-responsive element (CRE) DNA consensus sequences. We found that DOI induces a time-dependent increase in activator protein-1 and CRE transcription factor binding activity, which is blocked by 5-hydroxytryptamine2A receptor antagonists. Similar to 5 hydroxytryptamine2A receptor up-regulation, DOI-induced activator protein-1 binding is suppressed by inhibitors of calmodulin and Ca2+/calmodulin-dependent kinases. The increased activator protein-1 binding is effectively competed by excessive activator protein-1 and CRE sequences as well as endogenous activator protein-1-like sequences present in the rat 5-hydroxytryptamine2A receptor gene. Supershift assays revealed that cAMP-responsive element-binding protein (CREB) and Jun D are common components of both activator protein-1 and CRE binding complexes. DOI also increased the level of phospho-CREB in a time-dependent manner. The binding of phospho-CREB transcription factor to the activator protein 1 site suggests that CREB may modulate the transcription of genes that contain activator protein-1 but lack CRE site in their promoters, through interaction with the activator protein-1 site. The rat 5-hydroxytryptamine2A receptor up regulation may involve such a mechanism. PMID- 10363826 TI - Cellular and subcellular localization of 5-hydroxytryptamine1B receptors in the rat central nervous system: immunocytochemical, autoradiographic and lesion studies. AB - The localization of 5-hydroxytryptamine1B receptors in the rat central nervous system was investigated using anti-peptide antibodies that recognize a selective portion of the third intracytoplasmic loop of the receptor protein. At the light microscope level the densest 5-hydroxytryptamine1B receptor-like immunoreactivity was observed in ventral pallidum, globus pallidus, substantia nigra and dorsal subiculum. In addition, moderate immunoreactivity was found in the entopeduncular nucleus, the superficial gray layer of the superior colliculus, the caudate putamen and the deep nuclei of the cerebellum. This distribution matched perfectly that previously described from radioligand binding studies. At the ultrastructural level, 5-hydroxytryptamine1B receptor-like immunoreactivity was associated with axons and axon terminals in the three areas examined: substantia nigra, globus pallidus and superficial gray layer of the superior colliculus. In all cases, immunostaining was located on the plasma membrane of unmyelinated axon terminals and in the cytoplasm close to the plasmalemma. Synaptic differentiations were never labelled but, in some cases, 5-hydroxytryptamine1B receptor-like immunoreactivity was found in their close vicinity. Injection of kainic acid into the neostriatum resulted in a marked decrease in receptor-like immunoreactivity in the globus pallidus and the substantia nigra, consistent with the location of 5-hydroxytryptamine1B receptors on terminals of striatopallidal and striatonigral fibres, respectively. A reduction in 5-hydroxytryptamine1B receptor-like immunoreactivity was also noted in the superficial gray layer of the superior colliculus after contralateral enucleation, as expected of the location of 5-hydroxytryptamine1B receptors on the terminals of retinocollicular fibres. In both lesion experiments, immunolabelled degenerating terminals were observed in the projection areas. Anterograde labelling experiments coupled with immunocytochemical detection further showed that 5-hydroxytryptamine1B receptors in the substantia nigra are located on axons of striatal neurons. These data provide anatomical support for the idea that 5-hydroxytryptamine1B receptors act as terminal receptors involved in presynaptic regulation of the release of various neurotransmitters, including 5-hydroxytryptamine itself. PMID- 10363827 TI - Regulation of the LIM-type homeobox gene islet-1 during neuronal regeneration. AB - Peripheral nerve lesion leads to prominent changes in gene expression in the injured neurons, a process co-ordinated by transcription factors. During development the transcription factor islet-1 plays an important role in differentiation and axogenesis. In axotomized adult neurons a process of axonal regrowth and re-establishment of the neuronal function has to be activated. Thus, we studied changes in the expression of islet-1 after axotomy, under the assumption that frequently developmentally regulated factors are reactivated during neuronal regeneration. We investigated the regulation of islet-1 expression with (i) semi-quantitative reverse transcription polymerase chain reaction and (ii) confocal microscopy in combination with quantitative image analysis. Islet-1 expression was suprisingly down-regulated in motoneurons and sensory neurons of adult rats after axotomy. A maximal reduction in the expression level was reached between day 3 and 7 after nerve lesion, a period of extensive axonal sprouting. Islet-1 expression attained control level at day 42 after lesion, a time-point at which target reinnervation takes place. The decreased expression of islet-1 during axonal regeneration is in contrast to the high levels of islet-1 expression during axogenesis in the developing nervous system. Thus, the proposed role of islet-1 in axonal target finding during axogenesis could not be confirmed in the adult rat. The observed down-regulation of islet-1 rather suggests that the activation of downstream genes important for the embryonic pattern of axonal path finding is suppressed. Moreover, in the adult nervous system islet-1 might be one of the transcription factors regulating the expression of proteins significant for the physiological intact neuronal phenotype. PMID- 10363828 TI - Distribution of cholinergic neurons in cell group K of the rabbit brainstem. AB - The cell bodies of efferent neurons supplying the masseter and digastric muscles of the rabbit are located in two brainstem nuclei: the trigeminal motor nucleus and cell group k. The latter also contains neurons innervating muscles of the middle ear and Eustachian tube, as well as neurons that project to the cerebellum and the oculomotor complex. As part of an attempt to identify the functional subpopulations within the three cell divisions (kl-k3) that make up cell group k, we have investigated the distribution of neurons containing choline acetyltransferase, because these are likely to be motoneurons. Five rabbits anaesthetized with sodium pentobarbital (90 mg/kg, i.v.) were used in this study. They were perfused with 4% paraformaldehyde and 0.1% glutaraldehyde in phosphate buffer (0.1 M, pH 7.4). Two animals were used for preliminary studies. In the other three cases, serial Vibratome coronal sections of the brainstem were cut at 50 microm and two series of alternating sections were collected. The first was stained with a monoclonal antibody (code AB8, Incstar) directed against choline acetyltransferase, using the avidin-biotin-peroxidase method. The other was stained with Cresyl Violet. Cell counts and three-dimensional reconstructions were made for both series to determine positions and ratios of cholinergic and non-cholinergic neurons within the trigeminal motor nucleus and the subdivisions of cell group k. The results showed that the numbers of choline acetyltransferase and Nissl-stained neurons within the trigeminal motor nucleus were almost identical. In cell group k, significantly fewer choline acetyltransferase-stained cells were counted in all three animals (ratios of choline acetyltransferase/Nissl=0.53-0.71). In addition, the distribution of cholinergic neurons was not uniform throughout cell group k. Subdivisions kl and k3 contained proportionately fewer choline acetyltransferase-positive cells (ratios of choline acetyltransferase/Nissl=0.23-0.64) than did k2 (ratios choline acetyltransferase/ Nissl=0.75-0.88). Within each subdivision, there were significant differences in the spatial coordinates of Nissl- and choline acetyltransferase-positive neurons. We conclude that cell group k contains at least two populations of neurons which are unevenly distributed between and within the three subdivisions. While the majority of neurons in subgroup k2 contain choline acetyltransferase and thus are likely to be motoneurons, more than half of the neurons in subgroups k1 and k3 are not cholinergic. It remains to be determined whether these are the neurons that project to the cerebellum and to other CNS regions. PMID- 10363829 TI - Projections of medullary and pontine noradrenergic neurons to the horizontal limb of the nucleus of diagonal band in the rat. AB - Recent investigations in the rat have implicated a noradrenergic innervation to the horizontal nucleus of the diagonal band of Broca as a critical link in a neural circuit that conveys baroreceptor information centrally to inhibit the firing of vasopressin-secreting neurons in the hypothalamic supraoptic nucleus. In this study we used small intra-diagonal band injections of a retrograde tracer, rhodamine latex microspheres, in combination with tyrosine hydroxylase histochemistry to identify brainstem noradrenergic cells contributing to this innervation. In three cases where tracer injections were limited to the horizontal limb of the diagonal band, we observed 20-50 double-labelled neurons ipsilaterally in the dorsal part of the locus coeruleus (A6) and the caudal nucleus tractus solitarius (A2), and bilaterally in the caudal ventrolateral medulla (A1). Double-labelled neurons were also noted in the ventral tegmental area (dopaminergic A10 cell group). Although all major brainstem noradrenergic cell groups contribute fibers to the horizontal limb of the nucleus of diagonal band, data from physiological studies suggest that the noradrenergic A2 neurons in the nucleus tractus solitarius are the most likely pathway through which it receives this baroreceptor information. PMID- 10363830 TI - Identification of an efferent projection from the paraventricular nucleus of the hypothalamus terminating close to spinally projecting rostral ventrolateral medullary neurons. AB - The paraventricular nucleus of the hypothalamus is increasingly being viewed as an important site for cardiovascular integration because of its connections to regions in the brain and spinal cord which are known to be important in cardiovascular control. Like the vasomotor neurons of the rostral ventrolateral medulla, descending axons from paraventricular neurons can be identified that form synapses on sympathetic preganglionic neurons in the thoracic spinal cord. The purpose of this study was to determine whether paraventricular axons project to the rostral ventrolateral medulla and whether they are closely apposed to reticulospinal neurons in this region. Descending paraventricular axons were labelled with biotin dextran amine, while rostral ventrolateral medullary neurons were retrogradely labelled from the spinal cord with wheatgerm agglutinin conjugated to horseradish peroxidase. This revealed, within the rostral ventrolateral medulla, paraventricular axon and terminal varicosities closely apposed to and apparently contiguous with retrogradely labelled spinally projecting neurons. Thus our study at the light microscopical level has shown the potential for the paraventricular nucleus to directly influence rostral ventrolateral reticulospinal neurons. We suggest these connections, if confirmed by electron microscopy, could be one means by which activation of paraventricular neurons elicits alterations in blood pressure. PMID- 10363831 TI - Effect of graded spinal cord compression on cardiovascular neurons in the rostro ventro-lateral medulla. AB - In patients with spinal cord injury, cardiovascular disturbances such as hypotension, bradycardia and autonomic dysreflexia can be directly linked to abnormalities of central autonomic control. To date, the changes in bulbospinal innervation of sympathetic preganglionic neurons after compressive spinal cord injury have not been investigated. Thus, we examined the effect of varying severity of compressive spinal cord injury on neurons of the rostro-ventro lateral medulla, a nucleus of key importance in cardiovascular control. Adult rats with 20 g, 35 g and 50 g clip compression injuries (n= 18) of the cord at T1 and uninjured controls (n=13) were studied. Neurons in the rostro-ventro-lateral medulla with preserved spinal connections eight weeks after spinal cord injury were identified by retrograde labelling with 4% FluoroGold introduced into the cord at T6. Bulbospinal neurons in the rostro-ventro-lateral medulla were also examined immunocytochemically for the adrenaline-synthesizing enzyme phenylethanolamine-N-methyltransferase. In control rats an average of 451+/-12 rostro-ventrolateral medulla neurons were phenylethanolamine-N-methyltransferase positive. Of these, 213+/-6 projected to the T6 spinal cord. The number of rostro ventro-lateral medulla neurons retrogradely labelled by FluoroGold decreased as a linear function of severity of spinal cord injury (r= -0.95; P<0.0001). After 50g spinal cord injury at T1, only 7+/-1 rostro-ventro-lateral medulla neurons were labelled by FluoroGold, of which 6+/-1 were phenylethanolamine-N methyltransferase positive. Moreover, the number of phenylethanolamine-N methyltransferase positive rostro-ventro-lateral medulla neurons decreased to 361+/-16 after 50 g spinal cord injury. We conclude that compressive spinal cord injury results in disconnection of rostro-ventro-lateral medulla neurons, which project to the thoracic spinal cord, and that these changes vary with the severity of injury. The majority of these axotomized rostro-ventro-lateral medulla neurons maintain their immunopositivity for the adrenaline-synthesizing enzyme phenylethanolamine-N-methyltransferase. PMID- 10363833 TI - Identification of break points in mutated PMP22 gene in a new Trembler (Tr-Ncnp) mouse. PMID- 10363832 TI - Physiological and morphological correlates of presynaptic inhibition in primary afferents of the lamprey spinal cord. AB - Patch-clamp recordings in a whole-cell mode were performed on dorsal sensory cells enzymatically isolated from the spinal cord of two lamprey species, Ichthyomyzon unicuspis and Lampetra fluviatilis. The voltage-activated currents through calcium channels were analysed. GABA and the specific GABA(B) receptor agonist baclofen reduced the peak amplitude of inward Ba2+ current, as a robust alternate charge carrier through voltage-dependent Ca2+ channels. These effects were dose-dependent and reversible. GABA(B) receptor antagonists, 2 hydroxysaclofen and delta-amino-n-valeric acid, blocked the reduction of Ba2+ currents by GABA and baclofen, while bicuculline, a GABA(A) receptor antagonist, had no blocking action. GABA and baclofen did not modify the dorsal sensory cell membrane conductance, indicating that they did not activate ligand-gated channels. However, GABA, but not baclofen, considerably increased membrane conductance and induced Cl- currents in isolated multipolar neurons (presumably interneurons and/or motoneurons). These findings suggest that GABA and baclofen action on lamprey dorsal sensory cells is mediated by GABA(B) receptors. We concluded that GABA-mediated presynaptic inhibition of lamprey dorsal sensory cell fibers results from GABA(B) receptor activation followed by a decrease of inward voltage-activated calcium currents. Appositions of GABA-immunoreactive boutons to horseradish peroxidase-labeled fibers from the dorsal root were observed at the ultrastructural level in the dorsal column using postembedding immunogold cytochemistry. It seems likely that these appositions represent the morphological substrate of dorsal sensory cell fiber presynaptic inhibition. In very rare cases, ultrastructural features were observed which could be interpreted as synaptic specializations between the GABA-immunoreactive boutons and the primary afferent fibers. The extrasynaptic action of GABA as a basis of presynaptic inhibition of this population of primary afferent neurons is discussed. PMID- 10363834 TI - Traditional medicine in Turkey IX: folk medicine in north-west Anatolia. AB - Folk medicine in northwest Anatolia has been studied and 116 remedies prepared from 67 plant and 8 animal species are described, each with vernacular names, methods of preparation and traditional uses. PMID- 10363835 TI - Hypoglycaemic response to Zygophyllum gaetulum extracts in patients with non insulin-dependent diabetes mellitus. AB - Zygophyllum gaetulum leaves are one of several traditional remedies used for diabetes treatment in Morocco. Its ability to lower the blood glucose was studied in 13 patients with non-insulin-dependent diabetes mellitus. The reaction time of the Z. gaetulum aqueous extract at a single oral dose (440 mg/kg) producing a significant hypoglycaemic effect is 6 h after administration. The same dose ingested twice daily resulted in a significant reduction of blood glucose during the first week, and maintained the patient in normoglycaemia throughout the 2 week course of treatment, with no change in body weight. PMID- 10363836 TI - Effects of piperine on testis of albino rats. AB - Piperine was administered to mature male albino rats at doses of 5 and 10 mg/kg body weight, p.o., respectively, for 30 days. Only a 10 mg dose of piperine treatment caused a significant reduction in the weights of testis and accessory sex organs. Histological studies revealed that piperine at a 5 mg dose caused partial degeneration of germ cell types, whereas at a 10 mg dose, it caused severe damage to the seminiferous tubule, decrease in seminiferous tubular and Leydig cell nuclear diameter and desquamation of spermatocytes and spermatids. Correlated to the structural changes, a fall in caput and cauda epididymal sperm concentrations was also evident. A 10 mg dose of piperine also caused a marked increase in serum gonadotropins and a decrease in intratesticular testosterone concentration, despite normal serum testosterone titres. PMID- 10363837 TI - Antiplasmodial activity of selected Sudanese medicinal plants with emphasis on Maytenus senegalensis (Lam.) Exell. AB - The antiplasmodial activity of plant extracts related to four families was tested on chloroquine sensitive strain 3D7 and chloroquine resistant strain Dd2 of Plasmodium falciparum. The methanolic extract of Harrisonia abyssinica (Simaroubaceae) inhibited Dd2 with IC50 value of 4.7 microg/ml, while in 3D7, the IC50 value was 10 microg/ml. Most of the plants from the family Meliaceae showed highly potent antiplasmodial activity against the two tested strains. Khaya senegalensis, Azadirachta indica and Trichilia emetica showed IC50 values less than 5 microg/ml. The methanolic extract of Annona squamosa (Annonaceae) leaves showed high antiplasmodial activity with IC50 values of 2 and 30 microg/ml on 3D7 and Dd2, respectively. While stem bark showed moderate activity with IC50 values of 8.5 and 120 microg/ml on Dd2. Maytenus senegalensis (Celastraceae) possessed IC50 values of 3.9 on 3D7, 10 microg/ml on Dd2 and had no effect on lymphocyte proliferation even at the highest tested concentration; the IC50 was greater than 100 microg/ml. Liquid-liquid separation of the methanolic extract of M. senegalensis revealed that the dichloromethane extract possessed an IC50 value of only 2.1 microg/ml. Column fractionation of dichloromethane extract gave four fractions and fraction two showed an IC50 value of 0.5 microg/ml. Preliminary phytochemical analysis of dichloromethane fraction revealed terpenoids and traces of phenolic principles but no alkaloid, tannins or flavonoids were detected. PMID- 10363838 TI - Antibacterial, antifungal and antiviral activity of propolis of different geographic origin. AB - Propolis samples from different geographic origins were investigated for their antibacterial (against Staphylococcus aureus and Escherichia coli), antifungal (against Candida albicans) and antiviral (against Avian influenza virus) activities. All samples were active against the fungal and Gram-positive bacterial test strains, and most showed antiviral activity. The activities of all samples were similar in spite of the differences in their chemical composition. In samples from the temperate zone, flavonoids and esters of phenolic acids are known to be responsible for the above mentioned activities of bee glue; tropical samples did not contain such substances but showed similar activities. Obviously, in different samples, different substance combinations are essential for the biological activity of the bee glue. It seems that propolis has general pharmacological value as a natural mixture and not as a source of new powerful antimicrobial, antifungal and antiviral compounds. PMID- 10363839 TI - Antimicrobial properties of alkamides present in flavouring plants traditionally used in Mesoamerica: affinin and capsaicin. AB - The bioactive amides affinin and capsaicin isolated respectively from Heliopsis longipes roots and Capsicum spp fruits, were assayed for activity against Escherichia coli, Pseudomonas solanacearum, Bacillus subtilis and Saccharomyces cerevisicae suspension cultures. The alkamide affinin inhibited growth of E. coli and S. cerevisiae at concentrations as low as 25 microg/ml. Higher concentrations of affinin were necessary to inhibit growth of P. solanacearum and B. subtilis. However. high concentrations of capsaicin only retarded the growth of E. coli and P. solanacearum, whereas growth of B. subtilis was strongly inhibited and that of S. cerevisiae was initially enhanced. Results are discussed in relation to previous reports concerning crude extract and to the molecular structures of the bioactive compounds. PMID- 10363840 TI - Antimalarial activity of extracts of Malaysian medicinal plants. AB - In vitro and in vivo studies revealed that Malaysian medicinal plants, Piper sarmentosum, Andrographis paniculata and Tinospora crispa produced considerable antimalarial effects. Chloroform extract in vitro did show better effect than the methanol extract. The chloroform extract showed complete parasite growth inhibition as low as 0.05 mg/ml drug dose within 24 h incubation period (Andrographis paniculata) as compared to methanol extract of drug dose of 2.5 mg/ml but under incubation time of 48 h of the same plant spesies. In vivo activity of Andrographis paniculata also demonstrated higher antimalarial effect than other two plant species. PMID- 10363841 TI - Anthelmintic activity of the latex of Ficus species. AB - The latex of some species of Ficus (Moraceae) has been traditionally used as vermifuge in Central and South America. It has been accepted that anthelmintic activity is due to a proteolytic fraction called ficin. In the present study, the anthelmintic activity of the latex of Ficus insipida Willd. and Ficus carica L. has been investigated in NIH mice naturally infected with Syphacia obvelata, Aspiculuris tetraptera and Vampirolepis nana. The latex of F. insipida, administered by intragastric route in doses of 4 ml/kg/day during three consecutive days, were effective in the removal of 38.6% of the total number of S. obvelata, being inexpressive in the removal of A. tetraptera (8.4%) and segments of V. nana (6.3%). The latex of F. carica, administered in doses of 3 ml/kg/day, during three consecutive days, was effective in the removal of S. obvelata (41.7%) and it did not produce significant elimination of A. tetraptera (2.6%) and V. nana (8.3%). The observed high acute toxicity with hemorrhagic enteritis, in addition to a weak anthelmintic efficacy, do not recommend the use of these lattices in traditional medicine. PMID- 10363842 TI - Effect of trikatu, an Ayurvedic prescription, on the pharmacokinetic profile of rifampicin in rabbits. AB - The effect of single and multiple doses of a herbal preparation trikatu, an Ayurvedic prescription, on the bioavailability and pharmacokinetics of rifampicin was studied in rabbits. Rabbits (n = 10) were administered a single dose of rifampicin (24 mg/kg, p.o.) alone or in combination with a single dose of trikatu (500 mg/kg, p.o.). The study had a cross over design with a washout period of 7 days. In the other study, six rabbits were administered a single dose of rifampicin (24 mg/kg, p.o.) before and after multiple doses of trikatu (500 mg/kg x 7d, p.o.). In both studies, blood samples were collected at 0, 0.5, 1, 1.5, 2, 4, 6, 9 and 12 h after drug administration and assayed for rifampicin. In animals treated with single dose of trikatu there was a significant decrease in the peak plasma concentration (Cmax) of rifampicin (P < 0.05). Multiple doses of trikatu also reduced the Cmax and delayed the Tmax of rifampicin although not to a statistically significant level. Other pharmacokinetic parameters of rifampicin were not significantly altered. Our results suggest that coadministration of trikatu does not influence the extent of bioavailability (AUC0-infinity) but reduces the rate of bioavailability (Cmax) of rifampicin. And this latter effect may reduce the efficacy of rifampicin therapy. PMID- 10363843 TI - Screening of medicinal plants from Trinidad and Tobago for antimicrobial and insecticidal properties. AB - Antibacterial activity in 51 extracts from 29 plant species currently used in traditional medicine in Trinidad and the neighbouring Caribbean islands was tested for by the agar dilution streak method using six bacteria: Escherichia coli, Pseudomonas aeruginosa. Salmonella tophimurium, Staphylococcus aureus, Staphylococcus epidermidis and Enterococcus faecalis. The extracts from eight of the plants tested showed significant activity against one or more micro-organisms and the most susceptible bacterium was Staphylococcus aureus. In the bioassays for toxicity towards the Aedes aegypti mosquito the most effective plant extracts were from Justicia pectoralis, Manihot utilissima and Stachytarpheta jamaicensis. PMID- 10363844 TI - Antimicrobial activity of extracts of herbal plants used in the traditional medicine of Jordan. AB - Petroleum ether, ethanol, butanol, and aqueous crude extracts of the whole aerial parts of nine plants exhibited variable degrees of antimicrobial activity against four bacterial and three fungal species. Methanol and hexane extracts did not show any activity. Compared with standard antibiotics, extracts had low to moderate activity. The activity spectrum is wide against gram-positive and negative bacteria as well as fungi tested. However, the butanol extracts at 4 mg/disc of Ononis spinosa (OS), Bryonia syriaca (BS) had high moderate antifungal activity against Aspergillus flavus, Fusarium moniliforme and Candida albicans relative to miconazole nitrate at 40 microg/disc. Furthermore, higher antibacterial activity was observed though low to moderate compared with streptomycin and very comparable with chloramphenicol. Cyclaman persicum (CP) petroleum ether extracts only exhibited pronounced antibacterial activity. PMID- 10363845 TI - Antibacterial and antifungal activity of small protein of Indigofera oblongifolia leaves. AB - Four fractions from the leaves of Indigofera oblongifolia were obtained on Sephadex G-25 column chromatography. A single bands of these fractions were detected on Poly-acrylamide SDS gel electrophoresis. An antibacterial action of small protein peptide was tested against Escherichia coli, Klebsiella aerogenes, Kl. pneumoniae. Staphyllococcus aureus, and Bacillus subtilis. Peptide 3 showed strong inhibitory activity against B. subtilis and Aspergillus niger but clear zone of inhibition was also noted against A. fumigatus. Peptide 4 showed significant inhibitory zone against Kl. pneumoniae, S. aureus, B. subtilis. A. fumigatus, A. niger and A. flavus. PMID- 10363846 TI - Screening for biological activity and chemical composition of Euodia borbonica var. borbonica (Rutaceae), a medicinal plant in Reunion Island. AB - Three simple coumarins; scoparone, limettin and psoralen have been isolated as major components from the leaves of Euodia borbonica var. borbonica (Rutaceae) together with xanthoxylin, a common phenolic compound in Rutaceae family. Their structures were elucidated through GC/MS and NMR studies. A minor furocoumarin, bergapten, was also detected in the extracts. Preliminary biological tests on ethanolic leaf extracts did not show any activity. PMID- 10363847 TI - Reduction of toxicity of marrow transplantation in children with Fanconi anemia. PMID- 10363848 TI - Role of nutritional supplement in sickle cell disease. PMID- 10363849 TI - Consistently investigating the inconsistent nature of neuroblastoma. PMID- 10363850 TI - Metastatic sites in stage IV and IVS neuroblastoma correlate with age, tumor biology, and survival. AB - PURPOSE: The goal of this study was to determine the incidence of metastatic sites in neuroblastoma and the extent to which metastatic sites correlate with age, tumor biology, and survival. PATIENTS AND METHODS: All 648 patients with stage IV and IVS neuroblastoma registered on Children's Cancer Group protocols 3881 and 3891 were analyzed. Metastatic site data were provided by treating institutions and reviewed in patients with central nervous system (CNS), intracranial, lung, or "other" metastases. RESULTS: The incidence of metastatic sites at diagnosis was 70.5% in bone marrow, 55.7% in bone, 30.9% in lymph nodes, 29.6% in liver, 18.2% in intracranial and orbital sites, 3.3% in lung, and 0.6% in CNS. Event-free survival (EFS) was decreased in patients with bone, bone marrow, CNS, intracranial/ orbital, lung, and pleural metastases, and improved in those with liver and skin metastases. In infants, MYCN amplification and unfavorable Shimada histopathology correlated with increased frequencies of bone and intracranial or orbital metastases. In older patients, MYCN amplification correlated with increased frequencies of intracranial or orbital, liver, and lung metastases. Multivariate analysis revealed that metastatic site is not an independent prognostic factor. CONCLUSIONS: Metastatic pattern in neuroblastoma differs with age and correlates with tumor biological features and EFS. These correlations could reflect changes in host or tumor biological features with age resulting in differences in metastatic capacity or tumor affinity for specific sites. PMID- 10363851 TI - Treatment results of advanced neuroblastoma with the first Japanese study group protocol. Study Group of Japan for Treatment of Advanced Neuroblastoma. AB - PURPOSE: To elucidate the efficacy of intensive induction and consolidation chemotherapy regimens (Study Group of Japan for Advanced Neuroblastoma [JANB] 85) for patients with advanced neuroblastoma aged 1 year or older. PATIENT AND METHODS: One hundred fifty-seven patients with newly diagnosed advanced neuroblastoma were entered into this study between January 1985 and December 1990. Eligible patients were 12 months old or older with stage III or IV disease. The patients first received six cyclic courses of intensive induction chemotherapy (designated regimen A1) consisting of cyclophosphamide (1,200 mg/m2), vincristine (1.5 mg/m2), tetrahydro-pyranyl Adriamycin (pirarubicin; 40 mg/m2), and cisplatin (90 mg/m2). The patients were further treated with three different consolidation protocols: 3-[(4-amino-2-methyl-5-pyrimidinyl)methyl]-1 (2-chloroethyl)-1-nitrosour ea, dacarbazine, and bone marrow transplantation. RESULTS: Overall survival rates for patients with stage III disease without reference to the consolidation protocols were 80.8%, 76.9%, and 66.3% at 2, 5, and 10 years, respectively. The overall survival rates for patients with stage IV disease were 58.8%, 34.4%, and 28.9% at 2, 5, and 10 years, respectively. There were no statistically significant differences between the three consolidation treatment groups. Patients who did not achieve complete remission (CR) with induction chemotherapy and surgery all died, suggesting that CR is essential for the cure of advanced neuroblastoma. The overall 5-year survival rate of the 24 patients with N-myc amplified stage III and IV disease was 33.3%, and the longest survival time of a relapse-free patient was 103 months. CONCLUSION: The intensive induction chemotherapy regimen used in this study may be of significant value in increasing the CR rate and survival for patients with N-myc amplified and nonamplified advanced neuroblastoma. PMID- 10363852 TI - Cyclophosphamide for the treatment of progressive low-grade astrocytoma: a Pediatric Oncology Group phase II Study. AB - PURPOSE: Results of a phase II trial of cyclophosphamide (CPM) for children with progressive low-grade astrocytoma are reported. PATIENTS AND METHODS: Fifteen patients with a median age of 39 months (range, 2 to 71) were included in this study. The tumors of 11 children were located in the optic pathway, hypothalamus, or thalamus. Four courses of intravenous CPM 1.2 g/m2 were administered every 3 weeks during the upfront window portion of this protocol. Subsequently, chemotherapy was to continue with CPM, vincristine, and carboplatin for 2 years. RESULTS: By study design, the first 14 patients were centrally reviewed after completion of the initial 4 CPM courses. Toxicity was primarily hematologic. One patients had a complete response, 8 had stable disease, and 5 had progressive disease (PD). The excessive number of children with PD prompted study closure. CONCLUSION: CPM as used in this protocol showed insufficient activity against astrocytoma to justify further patient accrual. PMID- 10363853 TI - A multi-institutional retrospective study of intracranial ependymoma in children: identification of risk factors. AB - PURPOSE: The goal of this multi-institutional retrospective study of children with intracranial ependymoma was to identify risk factors associated with unfavorable overall survival (OS) and event-free survival (EFS). PATIENTS AND METHODS: Clinical data, including demographics, tumor location, spread, histology, details of surgery, radiation treatment, and chemotherapy were collected. Clinical characteristics and univariate and multivariate analyses of risk factors for OS and EFS are presented. RESULTS: Eleven U.S. institutions contributed 83 patients treated from 1987 to 1991. The OS at 5 and 7 years was 57% and 46%, and EFS at 5 and 7 years was 42% and 33%. Patients 3 years of age or younger differed from the older group by more common infratentorial location, less common gross total resection (GTR), and postoperative use of chemotherapy rather than radiation. This younger group of patients had worse survival (P < 0.01) than the older age group. Other than young age, less than GTR and World Health Organization (WHO) II grade 3 histology were significant adverse risk factors for EFS in univariate and multivariate analyses. OS shared the same adverse risk factors except for histology in multivariate analysis, which was only of borderline significance (P = 0.05). Progression at the original tumor location, present in 89% of patients, was the major pattern of tumor recurrence. Adjuvant chemotherapy in the group older than 3 years or craniospinal radiation in M0 patients did not significantly change EFS. CONCLUSIONS: Adverse outcome in childhood intracranial ependymoma is related to age (3 years or younger), histology (grade 3), and degree of surgical resection (less than GTR). New approaches, particularly for local tumor control in younger patients, are needed to improve survival. PMID- 10363854 TI - Economic and resource utilization analysis of outpatient management of fever and neutropenia in low-risk pediatric patients with cancer. AB - PURPOSE: To measure resource allocation in outpatient management of fever and neutropenia in low-risk pediatric patients with cancer and its impact on their families. PATIENTS AND METHODS: A prospective clinical trial was conducted. Eligible patients received a single dose of intravenous (IV) antibiotics and were observed for several hours in clinic. Patients were randomly assigned to continue either IV or oral antibiotics and were seen daily as outpatients. Charges were calculated based on the number of resources used and Medicare/Medicaid reimbursement schedules. A questionnaire was used to measure the impact of outpatient treatment on the family. RESULTS: Seventy-three episodes of fever and neutropenia were studied. The median duration of treatment was 4 days. Eighty-six percent of the episodes were managed without hospitalization. The median calculated charge was $1840. The median calculated charge for patients receiving oral antibiotics was $1544 and was significantly less than the $2039 median charge for outpatients treated with IV antibiotics. The estimated charge for comparable inpatient treatment was $4503. Nearly all families preferred outpatient care, and few reported a loss of work hours or increased child care expenses. CONCLUSIONS: Outpatient treatment of low-risk episodes of fever and neutropenia is substantially less costly than inpatient care and is preferred by most families. PMID- 10363855 TI - Plasma homocysteine levels and folate status in children with sickle cell anemia. AB - PURPOSE: A sensitive inverse relationship between plasma homocysteine concentration and folate status has been demonstrated. Although children with sickle cell anemia (SCA) are at potential risk for folate deficiency, plasma homocysteine levels have not been reported in such patients. Therefore, a study was designed to assess plasma homocysteine levels as a marker of folate status. DESIGN: Plasma homocysteine concentrations were measured in 120 children with SCA (102 in steady state and 18 during an acute complication) who had never received supplemental folic acid. Folate status was directly assessed in 34 of these patients. RESULTS: Plasma homocysteine levels in the patients with SCA and control subjects were similar. The mean value +/- 1 SD was 5.8+/-2.5 micromol/L (range, 1.6 to 14.1 micromol/L) in the patients with SCA and 6.1+/-2.7 micromol/L (range, 1.7 to 15.3 micromol/L) in 73 pediatric control subjects. In a subpopulation of the study group (34 children), simultaneous serum folate, red cell folate, and total homocysteine concentrations were also measured. Their serum folate and red cell folate concentrations were normal: 12.4+/-10.0 nmol/L (range, 1 to 42 nmol/L) and 604+/-374.7 nmol/L (range, 205 to 1741 nmol/L), respectively. There was no correlation of plasma homocysteine concentration with various clinical or laboratory measures or with red cell folate concentration. CONCLUSION: Folate stores in children with SCA not receiving folic acid supplements are adequate despite an underlying hemolytic anemia. PMID- 10363856 TI - Comparing alloimmunization in preterm infants after transfusion of fresh unmodified versus stored leukocyte-reduced red blood cells. AB - PURPOSE: To compare the occurrence of red blood cell (RBC), platelet (PLT), and white blood cell (WBC) antibodies in preterm infants after transfusions. METHODS: A randomized, blinded trial was conducted in which preterm infants were transfused either with stored RBCs, prepared by prestorage leukocyte reduction and transfused throughout 42 days of storage to limit donor exposure (n = 18), or with fresh RBCs prepared without leukocyte reduction and transfused within 7 days after collection from as many donors as needed to guarantee freshness (n = 17). Nontransfused preterm infants of comparable birth weight were control subjects (n = 11). RESULTS: No RBC antibodies were detected in serial blood samples taken during the first 6 months of life. Similarly, no definite WBC antibodies were found, although weak reactivity was detected transiently in sera from two infants. Accordingly, RBC and WBC antibody production did not differ among groups. In all, 11% of the transfused the infants exhibited platelet antibodies: 14% of the infants given stored leukocyte-reduced RBCs and 7% of the infants given fresh nonleukocyte-reduced RBCs (difference not statistically significant). CONCLUSIONS: Preterm infants rarely produce antibodies to blood cell antigens after RBC transfusions, regardless of whether the exposure is to fresh unmodified RBCs from several donors or to stored leukocyte-reduced RBCs from a limited number of donors. Therefore, efforts to limit donor exposures or to remove WBCs from blood components cannot be justified simply for purposes of preventing alloimmunization in neonates. PMID- 10363857 TI - Giant platelet disorders in African-American children misdiagnosed as idiopathic thrombocytopenic purpura. AB - A retrospective chart review of six African-American children with a diagnosis of macrothrombocytopenias (MTCP) was performed to evaluate the accuracy of their diagnosis. The following was diagnosed in the six children with MTCP: Fechtner syndrome (two children), Sebastian syndrome (one child), and unnamed MTCP (three children). In five families, chronic idiopathic thrombocytopenic purpura (ITP) was diagnosed in the propositus, which resulted in therapy using steroids, intravenous immunoglobulin (IVIG), and in one case splenectomy. Bleeding symptoms were generally mild. All six patients had thrombocytopenia ranging from 10 to 125 x 10(9)/L with mean platelet volume of 8 to 20 fL. Bleeding times were abnormal in two of three patients, and platelet aggregation was abnormal in three of four patients tested. Bone marrow aspirates were reported as increased megakaryocytes in the three patients on whom the procedure was performed. Ultrastructural morphology of platelets and leukocytes was performed in all six patients demonstrating giant platelets in all six patients and leukocyte inclusions in three patients. Differentiating MTCP from the more common ITP can be difficult but important in avoiding unnecessary diagnostic studies and potentially harmful therapy associated with ITP. PMID- 10363858 TI - Fludarabine-based protocol for human umbilical cord blood transplantation in children with Fanconi anemia. AB - PURPOSE: A novel conditioning regimen of fludarabine monophosphate (FLM), anti-T lymphocyte globulin (ATG), and low-dose cyclophosphamide with no irradiation for human umbilical cord blood transplantation (HUCBT) for the treatment of Fanconi anemia (FA) is described. PATIENT AND METHODS: A 12-year-old girl with FA received a human umbilical cord blood transplant from a fully matched sibling donor. After the HUCBT, the patient was given granulocyte colony stimulating factor in combination with erythropoietin. Pretransplant conditioning consisted of FLM (30 mg/m2/d) from day -10 to day -5, cyclophosphamide (10 mg/kg/d) on day 7 and -6, and rabbit ATG (ATG-Frasenius, 10 mg/kg/d) from day -4 to day -1. Cyclosporin A (3 mg/kg/d) was administered from day -1 as graft-versus-host disease prophylaxis. Cord blood from a sibling donor was used as a source of hematopoietic stem cells. RESULTS: Engraftment was normal and sustained. The regimen was well tolerated with very mild toxicity and no major transplant related complications or >grade II graft-versus-host disease. Chimerism was 100% donor origin as determined by restriction fragment length polymorphism. CONCLUSIONS: It is possible to achieve sustained engraftment and only mild toxicity in FA after HUCBT with a conditioning regimen of FLM, ATG, and cyclophosphamide with no irradiation. These preliminary results with this novel conditioning protocol are encouraging and should be evaluated in a larger group of patients with FA undergoing HUCBT. PMID- 10363859 TI - Lymphoblastic lymphoma and excessive toxicity from chemotherapy: an unusual presentation for Fanconi anemia. PMID- 10363860 TI - Familial Evans syndrome: a report of an affected sibship. AB - PURPOSE: This report describes the clinical course of three siblings, all of whom had Evans syndrome in childhood. PATIENTS: The coexistence of autoimmune hemolytic anemia and thrombocytopenia, in the absence of a known underlying cause, led to the diagnosis of Evans syndrome in a 4-month-old girl and subsequently in her two brothers when they were 4 and 13 years old. RESULTS: The 4-month-old girl had a life-threatening relapsing course unresponsive to corticosteroids, intravenous gamma-globulin, thymectomy, and cyclophosphamide. She eventually responded to splenectomy. Her two brothers had milder disease that responded to corticosteroids. Cytogenetic analyses revealed the presence of a familial Y;15 translocation in all three children and their father. CONCLUSION: There are few reported cases of familial Evans syndrome, and they are usually associated with an inherited congenital abnormality. We report the unusual finding of three siblings with the disease and no known congenital abnormality. PMID- 10363862 TI - Bone mineral density in survivors of cancer in childhood. PMID- 10363861 TI - Evans syndrome: successful management with multi-agent treatment including intermediate-dose intravenous cyclophosphamide. PMID- 10363863 TI - Catheter-based reperfusion of unprotected left main stenosis during an acute myocardial infarction (the ULTIMA experience). Unprotected Left Main Trunk Intervention Multi-center Assessment. AB - The ULTIMA registry was a prospective, multicenter, international registry of 277 patients who underwent percutaneous coronary interventions of unprotected left main trunk stenosis. The 40 patients who underwent an emergency percutaneous left main intervention for acute myocardial infarction are the focus of this study. We compared the results of primary angioplasty with primary stenting, characterizing both the short-term (in-hospital) and long-term (12-month) outcomes. Of the 40 patients, 23 underwent primary angioplasty, whereas 17 underwent primary stenting. The angiographic success rate was an 88% for the cohort. The in hospital death or coronary artery bypass grafting rate was 65% for the entire group, 74% for the percutaneous transluminal coronary angioplasty group (PTCA), and 53% for the stent group (p = 0.2). The in-hospital death rate was 55% for the entire cohort, 70% for the PTCA group, and 35% for the stent group (p = 0.1). The 12-month rate of death or bypass surgery was 83% and 58% for the PTCA and stent groups, respectively (p = 0.047). The 12-month survival rate was 35% and 53% for the PTCA and stent groups, respectively (p = 0.18). Bypass surgery was required in 6 patients in the PTCA group and 2 patients in the stent group (p = 0.07). Patients undergoing percutaneous interventions for unprotected left main myocardial stenosis during an acute myocardial infarction are critically ill; an initial percutaneous revascularization approach appears feasible and may be the preferred revascularization strategy. Primary stenting was associated with improved clinical outcomes. PMID- 10363864 TI - Intravascular ultrasound assessment of the relation between early and late changes in arterial area and neointimal hyperplasia after percutaneous transluminal coronary angioplasty and directional coronary atherectomy. AB - Previous serial intravascular ultrasound (IVUS) analysis after percutaneous transluminal coronary angioplasty or directional coronary atherectomy showed (1) early (within 1 month) increase in arterial area, (2) late (1- to 6-month) decrease in arterial area, and (3) an increase in plaque area from immediately to 6 months after intervention. To further understand these findings, we used serial IVUS to study the relations between changes in arterial and plaque area during the follow-up period after coronary intervention. Serial IVUS was performed before intervention and immediately, 24 hours, 1 month, and 6 months after percutaneous transluminal coronary angioplasty (n = 35) or directional coronary atherectomy (n = 26) in 57 patients. Arterial, lumen, and plaque areas were measured at the lesion site with the smallest preintervention and follow-up lumen areas at all time points. The increase in plaque area in the first month after intervention was accompanied by an equal or greater increase in arterial area (r = 0.670, p <0.0001). There was a decrease in arterial area from 1 to 6 months after intervention, which correlated inversely with both the increase in plaque area (r = 0.434, p <0.0001) or arterial area (r = 0.515, p <0.0001) during the first month after intervention and directly with the 1- to 6-month increase in plaque area (r = 0.460, p <0.0001). Comparison of the late (1 to 6 months) and early (within 1 month) delta arterial versus delta plaque area regression lines suggested that the late decrease in arterial area was superimposed on the relation between delta arterial area and delta plaque area. These relations were especially strong in restenotic (vs nonrestenotic) lesions. The early increase and late decrease in stenosis arterial area and neointimal hyperplasia appear to be interrelated, especially in restenotic stenoses. PMID- 10363865 TI - Usefulness of subcutaneous low molecular weight heparin (ardeparin) for reduction of restenosis after percutaneous transluminal coronary angioplasty. AB - In addition to its anticoagulant effects, heparin is known to have antiproliferative effects on vascular smooth muscle cells. Ardeparin is a partially depolymerized (low molecular weight) heparin that has a longer half life than unfractionated heparin. Following successful coronary balloon angioplasty, 565 patients were randomized to treatment with twice-daily subcutaneous ardeparin 50 anti-Xa U/kg (low dose) or 100 anti Xa U/kg body weight (high dose), or placebo for 3 months. Follow-up angiography was performed in 415 patients at 4 months, or earlier if clinically indicated. Additionally, patients underwent treadmill exercise electrocardiography at 2 weeks and 4 months. This study was designed to test the hypothesis that 3 months of subcutaneous dosing of ardeparin would reduce angiographic restenosis after coronary balloon angioplasty. Ardeparin had no effect on the incidence of angiographic restenosis (prespecified definition: > or = 50% luminal diameter narrowing plus a loss of 50% of initial gain or absolute decrease of 20% of luminal diameter). Neither the mean luminal diameters nor mean percent diameter stenoses were different among the treatment groups before, after, or 4 months after balloon angioplasty. On exercise electrocardiography at 2 weeks and 4 months, patients in all treatment groups had similar exercise tolerance, incidence of angina, and frequency of ST depression. Thus, ardeparin treatment given subcutaneously for 3 months after successful balloon angioplasty does not reduce either angiographic or clinical measures of restenosis. PMID- 10363866 TI - Presence and significance of the left atrionodal connection during atrioventricular nodal reentrant tachycardia. AB - It has been suggested that the anatomic substrates of dual atrioventricular nodal pathways are likely to be the atrionodal connections. During atrioventricular nodal re-entrant tachycardia (AVNRT) or ventricular pacing (VP), an earliest retrograde atrial activation in the coronary sinus (CS) distal to the ostium (CS breakthrough) would suggest the presence of an exit from a left atrionodal connection. The aim of the study was to evaluate the incidence of such an atrial retrograde activation in the CS during AVNRT and VP. The retrograde atrial activation was recorded during typical AVNRT (38 patients, 27 women, mean age 44 +/- 18 years) by a multipolar catheter in the CS, a decapolar catheter in the His bundle position, and a deflectable quadripolar catheter along the tricuspid annulus anterior to the CS ostium. In 31 patients the retrograde atrial activation was recorded also during VP at a similar cycle length. A CS breakthrough was found in 18 patients during AVNRT (47%) and in 13 patients during VP (42%). Presence or absence of CS breakthrough was concordant between AVNRT and VP in 90% of the patients. A CS breakthrough, suggesting a left-sided atrionodal connection, is frequently recorded both during AVNRT and VP. In patients with a CS breakthrough pattern, the absence of correlation between the His bundle to the earliest CS retrograde atrial electrogram interval and AVNRT cycle length, or any other atrial activation times recorded in the posterior and anterior region of the Koch's triangle, would suggest that the left-sided atrionodal connection is a bystander during typical AVNRT. PMID- 10363867 TI - Hemodynamic responses during leg press exercise in patients with chronic congestive heart failure. AB - To increase muscle mass and strength in patients with chronic congestive heart failure (CHF), there is a need for implementing resistance exercises in exercise training programs. This study sought to assess the safety of rhythmic strength exercise with respect to left ventricular function in 9 patients with stable CHF, compared with 6 stable coronary patients with mild left ventricular dysfunction (control group). With use of right-sided catheterization, changes in left ventricular function were assessed during double leg press exercise at loads of 60% and 80% of maximum voluntary contraction. The exercise sessions lasted 14 minutes each, divided into work and recovery phases of 60/120 seconds. In CHF, during exercise at a 60% load, there was a significant increase in heart rate (mean +/- SEM 90 +/- 4 beats/min; p <0.05), mean arterial blood pressure (95 +/- 3 mm Hg; p <0.01), diastolic pulmonary artery pressure (20.2 +/- 2.7 mm Hg; p <0.01), and cardiac index (3 +/- 0.3 L/m2/min; p <0.05). Additionally, during leg press exercise at an 80% load, there was a significant decrease in systemic vascular resistance (1,086 +/- 80 dynes x s x cm(-5); p <0.001), an increased cardiac index (3.4 +/- 0.1; p <0.001), and left ventricular stroke work index (75 +/- 5 g x m/m2; p <0.01), suggesting enhanced left ventricular function. Compared with controls, in CHF the magnitude of changes in hemodynamic parameters during exercise, demonstrated at a 60% load, was significantly smaller (systemic vascular resistance: [mean] 1,613 --> 1000 vs 1472 --> 1,247 dynes x s x cm(-5); cardiac index: 2.4 --> 3 vs 2.8 --> 4.4 L/m2/min, and stroke work index: 60 --> 69 vs 114 --> 155 g x m/m2; p <0.05 each). Nevertheless, changes indicated an enhanced contractile function of the left ventricle in CHF. This study demonstrates stability of left ventricular function during resistance exercise in well-compensated CHF patients with optimal drug therapy, as well as the appropriateness of the chosen mode and intensity applied as these factors relate to cardiovascular stress. This conclusion cannot be extrapolated to patients with less well-compensated heart failure, or to more protracted resistance training. PMID- 10363868 TI - Prognostic significance of atrial fibrillation in patients at a tertiary medical center referred for heart transplantation because of severe heart failure. AB - Atrial fibrillation (AF) occurs frequently in advanced heart failure. The prognostic significance of AF remains controversial. To determine the relation of AF to survival in patients with advanced heart failure, 234 consecutive patients referred for heart transplantation evaluation from January 1993 to June 1996 were studied to determine the effect of AF on event-free survival (freedom from death, heart transplantation, or placement of a left ventricular assist device). Clinical characteristics of the study population included: age, 51 +/- 17 years; maximum exercise oxygen consumption, 14.2 +/- 5.3 ml/kg/min; left ventricular ejection fraction, 24 +/- 11%; pulmonary capillary wedge pressure, 23 +/- 9 mm Hg; and ischemic etiology, 52%. Medical therapy included: diuretics (86%), angiotensin-converting enzyme inhibitors (80%), digoxin (80%), and anticoagulation therapy (72%). Mean duration of follow-up was 1.1 +/- 1.0 years. Sixty-two patients (27.4%) had AF. One-year event-free survival of the study population was 48%. No difference in event-free survival between patients with and without AF was observed. Univariate predictors of decreased event-free survival included: (1) advanced New York Heart Association class; (2) higher pulmonary capillary wedge pressure; (3) lower cardiac index; (4) lower maximum exercise oxygen consumption; (5) use of inotropic therapy; and (6) greater pulmonary artery systolic pressure. By multivariate analysis, independent predictors of decreased event-free survival included advanced New York Heart Association class (p <0.002) and higher pulmonary capillary wedge pressure (p = 0.02). Thus, AF in patients with advanced heart failure is not associated with decreased event-free survival. PMID- 10363869 TI - Cardiac sympathetic activity as measured by myocardial 123-I metaiodobenzylguanidine uptake and heart rate variability in idiopathic dilated cardiomyopathy. AB - In patients with idiopathic dilated cardiomyopathy (IDC) the increased sympathetic activity owing to chronic congestive heart failure leads to an imbalance of cardiac autonomic tone, as reflected by decreased heart rate variability (HRV). Iodine-123-metaiodobenzylguanidine (123-I-MIBG), which has the same affinity for sympathetic nerve endings as norepinephrine, can be used to assess the integrity and function of the cardiac sympathetic nervous system. The aim of the present study was to measure cardiac sympathetic activity by assessing 123-I-MIBG uptake compared with HRV in patients with IDC. In 12 patients with IDC and mild to moderate heart failure, myocardial MIBG uptake was calculated from the myocardial (M) to left ventricular cavity (C) voxel values density ratio and the 123-I activity in a blood sample as a reference (= M/C ratio) using a double radionuclide study with 123-I-MIBG and technetium-99m-MIBI. To investigate the relation between myocardial MIBG uptake and HRV in time domain, the linear regression between the M/C ratio, a new scintigraphic parameter, and the mean RR interval or the HRV triangular index, respectively, was determined. A significant correlation between the M/C ratio and mean RR interval (r = 0.52; p = 0.016) or M/C ratio and HRV triangular index (r = 0.76; p = 0.003), respectively, was found. Thus, the significant correlation between the M/C ratio and HRV indicate that they are both suitable noninvasive methods for evaluating cardiac sympathetic activity in patients with IDC and, furthermore, favor the view that there is evidence of a relation between HRV and the disorder of the cardiac presynaptic sympathetic nerve endings as demonstrated by a reduced M/C ratio. PMID- 10363870 TI - Outcome of children with atrial septal defect considered too small for surgical closure. AB - There are few studies providing information on the natural course of hemodynamically insignificant atrial septal defect (ASD). To review the outcome of patients with secundum ASD, we retrospectively reviewed the charts of patients who had initially not been considered for surgical closure after age 1 year, and who had either a follow-up of at least 10 years or documented closure. Thirty patients, 22 females and 8 males, fulfilled our inclusion criteria. Mean age at diagnosis was 1.3 year and mean follow-up duration was 11.5 years. Seventeen patients had spontaneous closure of the ASD at a mean age of 8.4 years. There were 7 asymptomatic patients whose ASD was still patent at the last visit (mean age 14.1 years, mean follow-up 13.2), with defect dimensions on echocardiography ranging from 1 to 6 mm. The remaining 6 patients were considered to require surgical closure on the basis of an apparent increase in size of the ASD and secondary clinical and hemodynamic manifestations. These results (1) confirm that not all secundum ASDs need to be treated surgically because they can still spontaneously close past the age of 5, and (2) suggest that in a minority of cases the size of the defect could increase. PMID- 10363872 TI - Prognostic usefulness of repeated echocardiographic evaluation after acute myocardial infarction. TRACE Study Group. TRAndolapril Cardiac Evaluation. AB - The prognostic value of repeated echocardiographic measurement of left ventricular function after acute myocardial infarction was evaluated. We found that repeated measurements of wall motion index in survivors of acute myocardial infarction, with no reinfarction, provide important prognostic information about death and worsening of heart failure. PMID- 10363871 TI - Acute coronary syndromes: the clot is there. AB - Attempts to extract clot from an infarct-related coronary artery have underestimated the frequency and pathophysiologic significance of thrombus in acute coronary syndromes. The overwhelming body of evidence garnered over the last 20 years, especially the achievement of reperfusion (Thrombolysis In Myocardial Infarction trial grade 2 and 3 flow) with most of the newer thrombolytic agents in >80% of cases, incontrovertibly establishes a critical role for thrombus in acute occlusive coronary artery disease. PMID- 10363873 TI - Estimation of coronary flow reserve using the Thrombolysis In Myocardial Infarction (TIMI) frame count method. AB - A simple and readily available method of estimating coronary flow velocity reserve may have significant clinical value. With use of intracoronary adenosine we documented a very good correlation between coronary flow reserve values obtained with the Thrombolysis In Myocardial Infarction trial frame count method and the invasive Doppler wire (Flowire) technique. PMID- 10363874 TI - Coronary artery stent restenosis responds favorably to repeat interventions. AB - One hundred sixty patients who underwent a percutaneous intervention for treatment of in-stent restenosis were evaluated on clinical follow-up to determine the predictors of reintervention, and also to compare rotational atherectomy with repeat percutaneous transluminal coronary angioplasty (PTCA) for this condition. Current smoking and saphenous vein graft location were the independent predictors of target vessel revascularization (TVR), and there was no difference in the rate of TVR between rotational atherectomy and re-PTCA. PMID- 10363875 TI - Incidence of bleeding complications associated with abciximab use in conjunction with thrombolytic therapy in patients requiring percutaneous transluminal coronary angioplasty. AB - The use of abciximab after full-dose failed thrombolytics within 15 hours of acute myocardial infarction significantly increases the risk of major bleeding complications. PMID- 10363876 TI - Association of low-density lipoprotein oxidation to abnormal electrocardiographic late potentials. AB - In vivo oxidation of low-density lipoprotein is shown to be significantly related to another risk factor for coronary atherosclerosis, abnormal electrocardiographic late potentials, in clinically healthy pilots. Because both of these variables have been also associated with cardiac arrhythmogenic action, together they may improve the identification of patients at risk of ventricular tachyarrhythmias. PMID- 10363877 TI - Kinetics of oxygen uptake at onset of exercise related to cardiac output, but not to arteriovenous oxygen difference in patients with chronic heart failure. AB - Patients with chronic heart failure performed exercise tests to evaluate the relation of kinetics of oxygen uptake to cardiac output and arteriovenous oxygen difference at the onset of exercise. The kinetics of oxygen uptake are primarily determined by cardiac output; these kinetics are useful in evaluating exercise intolerance and cardiac output response during exercise. PMID- 10363878 TI - Three-dimensional echocardiography enhances the assessment of ventricular septal defect. AB - By 3-dimensional echocardiography, the location, relation to the aortic and tricuspid valve, and the size of the ventricular septal defect was assessed and compared with 2-dimensional echocardiography and intraoperative findings. We concluded that 3-dimensional echocardiography accurately assesses the anatomy of the ventricular septal defect, provides additional information, and can be considered a valuable preoperative diagnostic tool. PMID- 10363879 TI - Echocardiographic and morphologic characteristics of left atrial myxoma and their relation to systemic embolism. AB - We examined the relation between the echocardiographic morphology of cardiac myxoma and systemic embolism in 25 patients. Two distinct types of myxoma could be identified by echocardiography: round type characterized by solid and round shape with nonmobile surface (n = 13, 52%), and polypoid type characterized by soft and irregular shape with mobile surface (n = 12, 48%); multiple regression analysis revealed the polypoid type of tumor was the only independent predictor of systemic embolism (p = 0.0029). PMID- 10363880 TI - Coronary blood flow velocity in normal infants and young adults assessed by transthoracic echocardiography. AB - The present study has demonstrated that it is possible to measure blood flow of the coronary artery in children using transthoracic 2-dimensional Doppler echocardiography. This is the first study on the relations between coronary flow velocity and age and heart rate in normal subjects. PMID- 10363881 TI - Evaluation of carotid arterial plaques after endarterectomy for Helicobacter pylori infection. AB - Chronic inflammation is linked to atherosclerosis and Helicobacter pylori has been suggested to be an etiologic agent, although the evidence is equivocal. In this report, H. pylori was not detected by the polymerase chain reaction in atherosclerotic plaque from carotid endarterectomy samples. PMID- 10363882 TI - Vascular endothelial growth factor: promise or peril? PMID- 10363884 TI - Recommendation for cigarette smoking cessation. PMID- 10363883 TI - Anterograde coronary angiography in tetralogy of Fallot. PMID- 10363885 TI - Effect of exercise on bioprosthetic valve hemodynamics. PMID- 10363886 TI - Diagnostic power of the electrocardiogram. PMID- 10363887 TI - Optimal environment for pacemaker and implantable cardioverter-defibrillator lead extraction with a laser sheath. PMID- 10363888 TI - Clinical experience over 48 years with pheochromocytoma. AB - OBJECTIVE: To analyze the presentation, localization, surgical management, pathology, and long-term outcome of a large series of patients with pheochromocytomas. SUMMARY BACKGROUND DATA: There are several areas of controversy pertaining to pheochromocytomas. Although many studies report a higher rate of malignancy for extraadrenal pheochromocytomas than for adrenal pheochromocytomas, the number of patients with the former tumor are small and statistical analysis is lacking. There has also been recent debate as to whether microscopic features of the tumor may be predictive of future behavior. METHODS: From 1950 to 1998, the authors observed 108 pheochromocytomas in 104 patients. The outcome of these patients has been followed prospectively. The medical records of these patients were reviewed for data on the presentation, localization, surgical management, pathology, and outcome. Patient survival was analyzed using Kaplan-Meier survival distributions. RESULTS: This study included 66 female patients and 38 male patients. The average age at surgery was 42.3 years. Sporadic cases accounted for 84% of the patients; the other 16% had multiple endocrine neoplasia type 2, von Recklinghausen's disease, von Hippel Lindau disease, or Carney's syndrome. Of 64 adrenal tumors, 55 were initially considered benign, 6 had microscopic malignant features, and 3 had malignant disease. Mean patient follow-up was 12.6 years. To date, in five additional patients (none with microscopic disease) malignant disease developed (13% overall rate of malignancy). Recurrence occurred as late as 15 years after resection. Of 26 extraadrenal pheochromocytomas, 14 were initially considered benign, 8 had microscopic malignant features, and 4 had malignant disease. Thus, 46% of patients had either malignant disease or tumors with malignant features. Mean patient follow-up was 11.5 years. In one patient with benign disease and in one patient with malignant features, malignant disease developed (23% overall rate of malignancy). The difference in the rate of malignancy was not statistically significant between adrenal and extraadrenal pheochromocytomas. Patients with adrenal and extraadrenal pheochromocytomas also had similar rates of survival (p = NS). CONCLUSIONS: The data suggest that patients with extraadrenal pheochromocytomas have the same risk of malignancy and the same overall survival as patients with adrenal pheochromocytomas. Lifelong follow-up of these patients is mandatory. PMID- 10363889 TI - Analysis of regression of postoperative carotid stenosis from prospective randomized trial of carotid endarterectomy comparing primary closure versus patching. AB - BACKGROUND AND PURPOSE: Recurrent stenosis after carotid endarterectomy (CEA) has been reported to vary between a few percent and 30%. Regression of recurrent stenosis has been reported sporadically in the literature, but studies analyzing the factors affecting regression are lacking. This study analyzed factors affecting the regression of postoperative stenosis from a prospective randomized trial of CEA comparing primary closure (PC) versus patching. PATIENT POPULATION AND METHODS: Three hundred ninety-nine CEAs were randomized into three groups: 135 PCs, 135 polytetrafluoroethylene patch closures (PTFE), and 130 vein patch closures (VPC). Postoperative duplex ultrasounds were done at 1, 6, and 12 months, and then yearly. The subgroup of these CEAs that exhibited postoperative stenosis was followed for possible regression of the stenosis. Analyses of various risk factors were examined for possible association with regression of recurrent stenosis. Mean follow-up was 46 months. RESULTS: Of 105 postoperative stenoses, regression was noted in 6/64 (9%) in PC, 6/13 (46%) in PTFE, and 10/28 (36%) in VPC. Overall, 22 recurrent stenoses regressed; 19 regressed to normal and 3 regressed from 50% to 80% stenosis to 20% to <50% stenosis. The mean time to regression was 383 days. Regression was more common in patching than PC. Both VPC and PTFE had significantly more regression than PC. When stenoses of 50% to 80% were analyzed, patching had more regression than PC. None of the recurrent stenoses > or = 80% regressed. There was no association between regression and other factors, including gender, hypertension, diabetes mellitus, coronary artery disease, smoking, internal carotid artery diameter, hyperlipidemia, hypercholesterolemia, or aspirin intake. CONCLUSIONS: Regression of recurrent stenosis was associated more strongly with patching than with PC. There was no association between regression and other factors. PMID- 10363890 TI - Evolution of the modified Rossetti fundoplication in children: surgical technique and results. AB - OBJECTIVE: To compare the modified Rossetti fundoplication with the classic Nissen. SUMMARY BACKGROUND DATA: The traditional surgical treatment of gastroesophageal reflux in children has been the classic Nissen fundoplication, defined by liver mobilization, crural repair, takedown of short gastric vessels, and floppy wrap. The authors have progressed in our technique of fundoplication and now perform a modified Rossetti fundoplication, defined by liver retraction without mobilization, no crural repair, short gastric vessels left intact, and 2 cm floppy wrap. METHODS: A retrospective chart review was performed on 407 pediatric patients who had open fundoplications (Jan. 13, 1993, to Feb. 25, 1998). Two groups were analyzed: the Nissen group (171 patients) and the Rossetti group (236 patients). Groups were compared for incidence of recurrent reflux, dysphagia, hiatal hernia, need for esophageal dilation, revision of fundoplication, time to discharge, and operative time. RESULTS: Incidence of dysphagia (3.7% vs. 3.3%), postoperative hiatal hernia (1.9% vs. 1.4%), need for esophageal dilation (1.2% vs. 0.5%), and need for fundoplication revision (2.5% vs. 2.3%) were similar between the groups. The mean operative time was significantly decreased in the Rossetti group (65 +/- 25 minutes) versus the Nissen group (73 +/- 33 minutes). Recurrent reflux occurred significantly more often in the Nissen group (11.2%) than in the Rossetti group (5.1 %). CONCLUSION: The modified Rossetti fundoplication has a low complication rate and is the authors' preferred method for the surgical treatment of gastroesophageal reflux in children. PMID- 10363891 TI - Percutaneous drainage of pancreatic pseudocysts is associated with a higher failure rate than surgical treatment in unselected patients. AB - OBJECTIVE: The primary aim was to compare directly the effectiveness of percutaneous drainage versus surgical treatment of pancreatic pseudocysts in unselected patients. The authors also wished to identify factors that may predict a successful outcome with percutaneous drainage. SUMMARY BACKGROUND DATA: Pancreatic pseudocysts are a common complication of pancreatitis, and recent data suggest that many pseudocysts may be observed or treated successfully by percutaneous drainage. Failures with percutaneous drainage have been recognized increasingly, and a direct comparison of percutaneous and surgical treatment was initiated to identify factors that may affect outcome with these approaches. METHODS: A computerized index search of the medical records of patients with a diagnosis of pancreatic pseudocyst was performed from 1984 to 1995. One hundred seventy-three patients were identified retrospectively and assigned to treatment groups: observation (n = 41), percutaneous drainage (n = 66), or surgical treatment (n = 66). Data on demographics, clinical presentation, pseudocyst etiology and characteristics, diagnostic evaluation, management, and outcome were obtained. Treatment failure was defined as persistence of a symptomatic pseudocyst or the need for additional intervention other than the original treatment. RESULTS: The etiology of pancreatitis, clinical presentation, and diagnostic evaluation did not differ between groups. Twenty-seven percent had documented chronic pancreatitis, and the etiology of pancreatitis was alcohol in 61% of patients. Mean pseudocyst size was 4.2 +/- 1 cm, 8.2 +/- 1.1 cm, and 7.4 +/- 1.3 cm in the observed, percutaneously treated, and surgically treated groups, respectively. Expectant treatment was successful in 93% of patients. Percutaneous drainage was successful in 42% of patients, whereas surgical treatment resulted in a success rate of 88%. Patients treated by percutaneous drainage had a higher mortality rate (16% vs. 0%), a higher incidence of complications (64% vs. 27%), and a longer hospital stay (45 +/- 5 days vs. 18 +/- 2 days) than patients treated by surgery. Eighty-seven percent of patients in whom percutaneous drainage failed required surgical salvage therapy. Multiple logistic regression analysis failed to reveal any factors significantly associated with a successful outcome after percutaneous drainage. CONCLUSIONS: Percutaneous drainage results in higher mortality and morbidity rates and a longer hospital stay than surgical treatment of pancreatic pseudocysts. The clinical benefit of percutaneous drainage of pancreatic pseudocysts in unselected patients has not been realized, and the role of this treatment should be established in a clinical trial. PMID- 10363892 TI - An analysis of 412 cases of hepatocellular carcinoma at a Western center. AB - OBJECTIVE: Using a large single-institution experience at a Western referral center, the authors examine partial hepatectomy as treatment of hepatocellular carcinoma and relate treatment outcomes to clinical parameters, including the etiology of underlying cirrhosis. METHODS: Four hundred and twelve patients seen between December 1991 and January 1998 were identified in a prospective database. Data about the surgical procedure, perioperative complications, and long-term outcome were examined. RESULTS: One hundred twenty-six patients did not have underlying cirrhosis. Of the 286 patients with cirrhosis, 119 were the result of hepatitis B, 39 hepatitis C, 36 both B and C, 43 ethanol abuse, and the remainder other causes. Two hundred forty-three patients underwent surgical exploration, and 154 patients underwent hepatic resection. Seven (4.5%) died from the surgery. One hundred forty-three patients were treated by ablative methods. Patients with cirrhosis had smaller tumors but nevertheless had a lower resectability rate. Neither the presence of cirrhosis nor the etiology of the cirrhosis altered the perioperative morbidity or mortality rate. The greatest determinant of long-term outcome was resectability. The size of the lesion, an alpha-fetoprotein level >2000 ng/ml, and vascular invasion were also determinants of poor outcome. The presence of cirrhosis was a detrimental factor when analysis was stratified for size of tumor. The cause of cirrhosis did not influence the long-term outcome. The 5-year survival rate was 57% for patients with resected lesions <5 cm and 32% for patients with tumors >10 cm. CONCLUSION: Partial hepatectomy is safe, effective, and potentially curative therapy for hepatocellular carcinoma. The presence of cirrhosis did not affect the surgical mortality rate but did affect the long-term survival rate. The cause of cirrhosis did not influence outcome. As treatment for small hepatocellular carcinomas, partial hepatectomy produces results similar to those of transplantation. For patients with large tumors who are poor candidates for transplantation, resection results in long-term survival in one third of patients. PMID- 10363893 TI - The epidemic of cocaine-related juxtapyloric perforations: with a comment on the importance of testing for Helicobacter pylori. AB - OBJECTIVE: This is a report of 50 consecutive patients with juxtapyloric perforations after smoking "crack" cocaine (cocaine base) at one urban public hospital. SUMMARY BACKGROUND DATA: Although the exact causal relation between smoking crack cocaine and a subsequent juxtapyloric perforation has not been defined, surgical services in urban public hospitals now treat significant numbers of male addicts with such perforations. This report describes the patient set, presentation, and surgical management and suggests a possible role for Helicobacter pylori in contributing to these perforations. METHODS: A retrospective chart review was performed, supplemented by data from the patient log in the department of surgery. RESULTS: From 1994 to 1998, 50 consecutive patients (48 men, 2 women) with a mean age of 37 had epigastric pain and signs of peritonitis a median of 2 to 4 hours (but up to 48 hours) after smoking crack cocaine. A history of chronic smoking of crack as well as chronic alcohol abuse was noted in all patients; four had a prior history of presumed ulcer disease in the upper gastrointestinal tract. Free air was present on an upright abdominal x ray in 84% of patients, and all underwent operative management. A 3- to 5-mm juxtapyloric perforation, usually in the prepyloric area, was found in all patients. Omental patch closure was used in 49 patients and falciform ligament closure in 1. Two patients underwent parietal cell vagotomy as well. In the later period of the review, antral mucosal biopsies were performed through the juxtapyloric perforation in five patients. Urease testing was positive for infection with H. pyonri in four, and these patients were prescribed appropriate antimicrobial drugs. CONCLUSIONS: Juxtapyloric perforations after the smoking of crack cocaine occur in a largely male population of drug addicts who are 8 to 10 years younger than the patient group that historically has perforations in the pyloroduodenal area. These perforations are usually 3 to 5 mm in diameter, and an antral mucosal biopsy for subsequent urease testing should be performed if the location and size of the ulcer allow this to be done safely. Omental patch closure is appropriate therapy for patients without a history of prior ulcer disease; antimicrobial therapy and omeprazole are prescribed when H. pylori is present. PMID- 10363894 TI - Paying a premium: how patient complexity affects costs and profit margins. AB - OBJECTIVE AND BACKGROUND: Tertiary medical centers continue to be under extreme pressure to deliver high-complexity care, but paradoxically there is considerable pressure within these institutions to reduce their emphasis on tertiary care and refocus their efforts to develop a more community-like practice. The genesis of this pressure is the perceived profitability of routine surgical activity when compared with more complex care. The purpose of this study is to assess how the total cost and profit (loss) margin can vary for an entire trauma service. The authors also evaluate payments for specific trauma-related diagnostic-related groups (DRGs) and analyze how hospital margins were affected based on mortality outcome. MATERIALS AND METHODS: The authors analyzed the actual cost of all trauma discharges (n = 692) at their level I trauma center for fiscal year 1997. Data were obtained from the trauma registry and the hospital cost accounting system. Total cost was defined as the sum of the variable, fixed, and indirect costs associated with each patient. Margin was defined as expected payments minus total cost. The entire population and all DRGs with 10 or more patients were stratified based on survival outcome, Injury Severity Score, insurance status, and length of stay. The mean total costs for survivors and nonsurvivors within these various categories and their margins were evaluated. RESULTS: The profit margin on nonsurvivors was $5,898 greater than for survivors, even though the mean total cost for nonsurvivors was $28,821 greater. Within the fixed fee arrangement, approximately 44% of transfers had a negative margin. Both survivors and nonsurvivors become increasingly profitable out to 20 days and subsequently become unprofitable beyond 21 days, but nonsurvivors were more profitable than survivors. CONCLUSIONS: There is a wide variance in both the costs and margins within trauma-related DRGs. The DRG payment system disproportionately reimburses providers for nonsurvivors, even though on average they are more costly. Because payers are likely to engage in portfolio management, patients can be transferred between hospitals based on the contractual relationship between the payer and the provider. This payment system potentially allows payers to act strategically, sending relatively low-cost patients to hospitals where they use fee-for-service reimbursement and high-cost patients to hospitals where their reimbursement is contractually capped. Although specific to the authors' trauma center and its payer mix, these data demonstrate the profitability of maintaining a level I trauma center and preserving the mission of delivering care to the severely injured. PMID- 10363895 TI - Prognosis and survival in patients with gastrointestinal tract carcinoid tumors. AB - OBJECTIVE: To determine the impact of clinical presentation variables on the management and survival of patients with gastrointestinal (GI) tract carcinoid tumors. METHODS: A 20-year (1975-1995) retrospective analysis of 150 patients with GI tract carcinoid tumors at the Massachusetts General Hospital was conducted. Median follow-up was 66 months (range 1-378). Survival estimates for prognostic factors were calculated using Kaplan-Meier product limit estimators, with death from carcinoid as the outcome. Univariate analyses for each factor were obtained using a log-rank test, and multivariate survival analysis was performed. RESULTS: All but two patients underwent surgical intervention with the intent to cure (90%) or debulk the tumor (9%). Mean age at presentation was 55 +/ 18 years (range 11-90). There was a slight female/male predominance (80:70). Symptoms were nonspecific; the most common were abdominal pain (40%), nausea and vomiting (29%), weight loss (19%), and GI blood loss (15%). Incidental carcinoids, discovered at the time of another procedure, occurred in 40% of patients and were noted at multiple sites throughout the GI tract. The distribution of tumors was ileojejunum (37%), appendix (31 %), colon (13%), rectum (12%), stomach (4%), duodenum (1.3%), and Meckel's diverticulum (1.3%). Of the 27 patients with documented liver metastases, carcinoid syndrome developed in only 13 patients (48%), manifested by watery diarrhea (100%), upper body flushing (70%), asthma (38%), and tricuspid regurgitation (23%). All 13 patients with carcinoid syndrome had elevated levels of 5-HIAA, but the absolute levels did not correlate with the severity of symptoms. An additional 11 patients, 3 without liver metastases, had elevated levels of 5-HIAA without any evidence of carcinoid syndrome. Multicentric carcinoid tumors occurred in 15 patients (10%), and all but one of these tumors were centered around the ileocecal valve. There was no difference in the incidence of liver metastases between solitary (18%) and multicentric carcinoids (20%). Synchronous noncarcinoid tumors were present in 33 patients (22%), and metachronous tumors developed in an additional 14 patients (10%) in follow-up. Age and tumor size, depth, and location were significant predictors of metastases. By multivariate analysis, age > or = 50 years, metastases, and male gender were statistically significant predictors of death. CONCLUSIONS: Gastrointestinal tract carcinoid tumors have a nonspecific clinical presentation, except in the case of the carcinoid syndrome. Surgical resection is the treatment of choice for improving survival. Surgically treated patients with carcinoid tumor have an overall favorable 83% 5-year survival rate. PMID- 10363896 TI - Orthotopic liver transplantation for hepatitis C: outcome, effect of immunosuppression, and causes of retransplantation during an 8-year single-center experience. AB - OBJECTIVE: To determine the outcome of orthotopic liver transplantation (OLT) for end-stage liver disease caused by hepatitis C virus (HCV). SUMMARY BACKGROUND DATA: HCV has become the leading cause of cirrhosis and hepatic failure leading to OLT. Recurrent HCV after OLT is associated with significant complications and may lead to graft loss that requires retransplantation (re-OLT). The authors studied the outcome of transplantation for HCV, the effect of primary immunotherapy, and causes of retransplantation. METHODS: The authors conducted a retrospective review of their experience during an 8-year period (1990-1997), during which 374 patients underwent transplants for HCV (298 [79.6%] received one OLT; 76 [20.4%] required re-OLT). Median follow-up was 2 years (range 0 to 8.3). Immunosuppression was based on cyclosporine in 190 patients and tacrolimus in 132 patients. In a third group of patients, therapy was switched from cyclosporine to tacrolimus or from tacrolimus to cyclosporine (cyclosporine/tacrolimus group). RESULTS: Overall, 1-, 2-, and 5-year actuarial patient survival rates were 86%, 82%, and 76%, respectively. The 2-year patient survival rate was 81 % in the cyclosporine group, 85% in the tacrolimus group, and 82% in the cyclosporine/tacrolimus group. In patients receiving one OLT, overall 1-, 2-, and 5-year patient survival rates were 85%, 81%, and 75%, respectively. The 2-year patient survival rate was 79% in the cyclosporine group, 84% in the tacrolimus group, and 80% in the cyclosporine/tacrolimus group. The overall graft survival rates were 70%, 65%, and 60% at 1, 2, and 5 years, respectively. The graft survival rate at 2 years was similar under cyclosporine (68.5%), tacrolimus (64%), or cyclosporine/tacrolimus (60%) therapy. Re-OLT was required in 42 (11.2%) patients for graft dysfunction in the initial 30 days after OLT. Other causes for re-OLT included hepatic artery thrombosis in 10 (2.6%), chronic rejection in 8 (2.1%), and recurrent HCV in 13 (3.4%) patients. The overall survival rates after re-OLT were 63% and 58% at 1 and 2 years. The 1-year survival rate after re-OLT was 61 % for graft dysfunction, 50% for chronic rejection, 60% for hepatic artery thrombosis, and 60% for recurrent HCV. At re OLT, 85.3% of the patients were critically ill (United Network for Organ Sharing [UNOS] status 1); only 14.7% of the patients were UNOS status 2 and 3. In re-OLT for chronic rejection and recurrent HCV, the 1-year survival rate of UNOS 1 patients was 38.4%, compared with 87.5% for UNOS 2 and 3 patients. In patients requiring re-OLT, there was no difference in the 1-year patient survival rate after re-OLT when cyclosporine (60%), tacrolimus (63%), or cyclosporine/tacrolimus (56%) was used for primary therapy. With cyclosporine, three patients (1.5%) required re-OLT for chronic rejection versus one patient (0.7%) with tacrolimus. Re-OLT for recurrent HCV was required in four (3%) and seven (3.6%) patients with tacrolimus and cyclosporine therapy, respectively. CONCLUSIONS: Orthotopic liver transplantation for HCV is performed with excellent results. There are no distinct advantages to the use of cyclosporine versus tacrolimus immunosuppression when patient and graft survival are considered. Re OLT is an important option in the treatment of recurrent HCV and should be performed early in the course of recurrent disease. Survival after re-OLT is not distinctively affected by cyclosporine or tacrolimus primary immunotherapy. The incidence of re-OLT for recurrent HCV or chronic rejection is low after either tacrolimus or cyclosporine therapy. PMID- 10363897 TI - Specific therapy for local and systemic complications of acute pancreatitis with monoclonal antibodies against ICAM-1. AB - OBJECTIVE: To analyze the time points and levels of the expression of adhesion molecules in the pancreas and lung in pancreatitis of different severities, and to assess whether treatment with a monoclonal antibody against intercellular adhesion molecule-1 (ICAM-1) can reduce local and systemic complications. BACKGROUND: The outcome of severe acute pancreatitis relates to its pulmonary and septic complications. Leukocyte adhesion and infiltration, both mediated by ICAM 1, are central events in the pathogenesis of necrotizing pancreatitis. METHODS: Expression of ICAM-1 at different time points was assessed by immunohistochemistry and Western blot analysis in pancreas and lungs from rats with mild edematous or severe necrotizing pancreatitis. ICAM-1 expression was correlated with leukocyte infiltration and histologic changes. The possible therapeutic effect of monoclonal antibodies against ICAM-1 was assessed by measuring pancreatic and lung injury. RESULTS: In edematous pancreatitis, increased ICAM-1 expression in pancreas was evident by 6 hours but did not occur in lung. In contrast, ICAM-1 was upregulated at 3 hours in the pancreas and at 12 hours in lung in necrotizing pancreatitis. Increased expression of ICAM-1 preceded leukocyte infiltration. Treatment of severe necrotizing pancreatitis with monoclonal antibodies against ICAM-1 decreased both local pancreatic injury and systemic lung injury compared with untreated controls. CONCLUSIONS: Upregulation of ICAM-1 and subsequent leukocyte infiltration appear to be significant mediators of pancreatic and pulmonary injury in pancreatitis, and both the onset and extent correlate with severity. The time course should permit effective prevention of tissue damage by treatment with ICAM-1 antibodies. PMID- 10363898 TI - Tolerance to shock: an exploration of mechanism. AB - OBJECTIVE: To determine if cross-tolerance to septic shock could be induced by a previous insult with sublethal hemorrhage (SLH) and to characterize the mechanisms involved in this induced protective response. BACKGROUND DATA: It is possible to condition animals by prior SLH such that they tolerate an otherwise lethal hemorrhage. It is also possible to condition animals with low doses of lipopolysaccharide (LPS) so that they survive a "lethal" septic insult. However, a paucity of information exists on cross-tolerance between hemorrhage and sepsis. METHODS: Rats were made tolerant by conditioning SLH or sham operation. Twenty four hours later, tolerant and sham rats were exposed to a lethal dose of LPS. To explore the mechanism of tolerance induction, rats were given the macrophage (Mphi) inhibitor CNI-1493 or saline carrier before SLH. Survival and pulmonary vascular injury were determined after LPS. Serum tumor necrosis factor (TNF) levels and splenic Mphi TNF gene expression were measured at several time points. RESULTS: Prior SLH indeed made rats tolerant and imparted a significant survival benefit and reduction in pulmonary vascular injury after LPS. The tolerance induced by SLH was reversed by Mphi inhibition. Tolerant animals had low serum TNF levels immediately after SLH and reduced circulating TNF levels after LPS. SLH, however, did not inhibit the augmentation of TNF gene expression after LPS. CONCLUSIONS: Sublethal hemorrhage bestows protection against a lethal LPS challenge. Inhibition of the Mphi attenuated the benefit of the tolerance induced by SLH. Circulating TNF but not TNF gene after LPS is lessened by SLH. This implicates changes in Mphi intracellular signaling in induction of the tolerant state. PMID- 10363899 TI - Caspase-3-dependent organ apoptosis early after burn injury. AB - OBJECTIVE: To examine the role played by endotoxin, tumor necrosis factor-alpha (TNF-alpha), and caspase-3 in the increased apoptosis seen in solid organs in the early period after a burn injury. SUMMARY BACKGROUND DATA: Burn injury is often associated with immune suppression. Bacterial translocation and systemic endotoxemia have been reported after a burn injury, and caspase-3 activation due to TNF-alpha and Fas ligand (FasL) are presumed to initiate apoptosis. We hypothesized that endotoxin-induced TNF-alpha expression and caspase-3 activation could be the stimulus for the apoptosis after burn injury. METHODS: A 20% full thickness scald burn was used in C57BL/6 mice. Three hours after burn injury, tissue samples were obtained from the thymus, lung, liver, and spleen. Lipopolysaccharide-nonresponsive (C3H/HeJ) and TNFalpha null B6x129tnf-/- mice were also used. To detect apoptosis, hematoxylin and eosin stain, in situ TUNEL, DNA extraction, and gel electrophoresis were all performed. Caspase-3 activity and TNF-alpha and FasL mRNA were also measured. RESULTS: Increased apoptosis and caspase-3 activity were observed in the thymus and spleen 3 hours after burn injury but were not seen in liver or lung. In the thymus and spleen, increased expression of FasL mRNA was also observed, whereas increased TNF-alpha mRNA was not. Increased apoptosis in thymus and spleen were also observed in C3H/HeJ and B6x129tnf-/- mice after burn injury. An inhibitor of the caspase-3 (Z-VAD-fmk) reduced apoptosis in both thymus and spleen. CONCLUSIONS: In the early period after a burn injury, increased apoptosis is observed primarily in the lymphoid organs and is independent of endotoxin or TNF-alpha. The increased caspase-3 activity in thymus and spleen contributes to apoptosis in these organs. PMID- 10363900 TI - Subareolar versus peritumoral injection for location of the sentinel lymph node. AB - BACKGROUND: Sentinel lymph node (SLN) biopsy is fast becoming the standard for testing lymph node involvement in many institutions. However, questions remain as to the best method of injection. The authors hypothesized that a subareolar injection of material would drain to the same lymph node as a peritumoral injection, regardless of the location of the tumor. METHODS: To test this theory, 68 patients with 69 operable invasive breast carcinomas and clinically node negative disease were enrolled in this single-institution Institutional Review Board-approved trial. Patients were injected with 1.0 mCi of technetium-99 sulfur colloid (unfiltered) in the subareolar area of the tumor-bearing breast. Each patient received an injection of 2 to 5 cc of isosulfan blue around the tumor. Radioactive SLNs were identified using a hand-held gamma detector probe. RESULTS: The average age of patients entered into this trial was 55.2 +/- 13.4 years. The average size of the tumors was 1.48 +/- 1.0 cm. Thirty-two percent of the patients had undergone previous excisional breast biopsies. Of the 69 lesions, 62 (89.9%) had SLNs located with the blue dye and 65 (94.2%) with the technetium. In four patients, the SLN was not located with either method. All blue SLNs were also radioactive. All located SLNs were in the axilla. Of the 62 patients in which the SLNs were located with both methods, an average of 1.5 +/- 0.7 SLNs were found per patient, of which 23.2% had metastatic disease. All four patients in which no SLN was located with either method had undergone prior excisional biopsies. CONCLUSIONS: The results of this study suggest that subareolar injection of technetium is as accurate as peritumoral injection of blue dye. Central injection is easy and avoids the necessity for image-guided injection of nonpalpable breast lesions. Finally, subareolar injection of technetium avoids the problem of overlap of the radioactive zone of diffusion of the injection site with the radioactive sentinel lymph node, particularly in medial and upper outer quadrant lesions. PMID- 10363901 TI - The enigma of desmoid tumors. AB - OBJECTIVE: To analyze patients with recurrent extremity desmoids, in whom the surgical therapeutic option was either major amputation or observation. SUMMARY BACKGROUND DATA: The biology and natural history of desmoid tumors are an enigma. These tumors invade surrounding structures and recur locally but do not metastasize. The morbidity of treating these tumors in the context of their relatively benign biology is uncertain. METHODS: Between July 1982 and June 1998, the authors treated and prospectively followed 206 patients with extremity desmoid tumors. All patients underwent standardized surgical resection, the surgical goal always being complete resection with negative margins. When tumors recurred, they were evaluated for reresection. Amputation was considered when resection was not possible because of neurovascular or major bone involvement, or in the presence of a functionless, painful extremity. RESULTS: During this period, 22 patients had disease that was not resectable without amputation. This was out of a total of 115 patients with primary disease and 91 patients with recurrent disease. All recurrences were local; in no patient did metastasis develop. In this group of 22 patients with unresectable disease, 7 underwent amputation and 15 did not. These 15 patients were followed, alive with disease, having no surgical resection. Four patients received systemic treatment with tamoxifen and nonsteroidal antiinflammatories, three received systemic cytotoxic chemotherapy, and two received both tamoxifen and chemotherapy. Six patients received no systemic treatment. The range of follow-up was 25 to 92 months. In all patients, there was no or insignificant tumor progression; in three patients who underwent observation alone, there was some regression of tumor. During follow-up, no patient has required subsequent amputation, and no patient has died from disease. CONCLUSIONS: In desmoid tumors, aggressive attempts at achieving negative resection margins may result in unnecessary morbidity. Function- and structure-preserving procedures should be the primary goal. In select patients, whose only option is amputation, it may be prudent to observe them with their limb and tumor intact. PMID- 10363902 TI - Improved success rate in reoperative parathyroidectomy with intraoperative PTH assay. AB - OBJECTIVE: The clinical usefulness of preoperative localization and intraoperative PTH assay (QPTH) in primary hyperparathyroidism have been established. However, without the use of QPTH, the parathyroidectomy failure rate remains 5% to 10% in large reported series and is probably much higher in the hands of less experienced parathyroid surgeons. Persistent hypercalcemia requires another surgical procedure. The authors compared the outcomes in 50 consecutive patients undergoing more difficult secondary parathyroidectomy with and without the adjunctive support of QPTH. METHODS: Two groups of similar patients underwent reoperative parathyroidectomy for failed surgery or recurrent disease. The successful return to normocalcemia in group I, with QPTH used to localize and confirm complete excision of all hyperfunctioning glands, was compared with group II, who did not have this intraoperative adjunct. RESULTS: In 31/33 patients in group I, calcium levels returned to normal. With good preoperative localization studies, 17 patients underwent successful straightforward parathyroidectomies as predicted by QPTH. In the other 14 patients, QPTH assay proved extremely beneficial by facilitating localization with differential venous sampling; measuring the increase in hormone secretion after massage of specific areas; recognizing suspicious nonparathyroid tissue excised without a decrease in hormone levels, avoiding frozen-section delay; and correctly identifying the excision of abnormal tissue despite false-positive/false-negative sestamibi scans. In group II, who underwent surgery before QPTH was available, 4 of 17 patients (24%) remained hypercalcemic after extensive reexploration. CONCLUSION: With the intraoperative hormone assay used to facilitate localization and confirm excision of all hyperfunctioning tissue, the success rate of reoperative parathyroidectomy has improved from 76% to 94%. PMID- 10363903 TI - Patterns of nodal metastases in palpable medullary thyroid carcinoma: recommendations for extent of node dissection. AB - OBJECTIVE: To establish the frequency, pattern and location of cervical lymph node metastases from palpable medullary thyroid carcinoma (MTC). Recommendations are made regarding the extent of surgery for this tumor. SUMMARY BACKGROUND DATA: Medullary thyroid carcinoma is a tumor of neuroendocrine origin that does not concentrate iodine. Surgical extirpation of the thyroid tumor and cervical node metastases is the only potentially curative therapeutic option. Patterns of node metastases in the neck and guidelines for the extent of dissection for palpable MTC are not well established. METHODS: Seventy-three patients underwent thyroidectomy for palpable MTC with immediate or delayed central and bilateral functional neck dissections. The number and location of lymph node metastases in the central (levels VI and VII) and bilateral (levels II to V) nodal groups were noted and were correlated with the size and location of the primary thyroid tumor. Intraoperative assessment of nodal status by palpation and inspection by the surgeon was correlated with results of histologic examination. RESULTS: Patients with unilateral intrathyroid tumors had lymph node metastases in 81% of central node dissections, 81% of ipsilateral functional (levels II to V) dissections, and 44% of contralateral functional (levels II to V) dissections. In patients with bilateral intrathyroid tumors, nodal metastases were present in 78% of central node dissections, 71% of functional (levels II to V) node dissections ipsilateral to the largest intrathyroid tumor, and 49% of functional (levels II to V) node dissections contralateral to the largest thyroid tumor. The sensitivity of the surgeon's intraoperative assessment for nodal metastases was 64%, and the specificity was 71%. CONCLUSION: In this series, >75% of patients with palpable MTC had associated nodal metastases, which often were not apparent to the surgeon. Routine central and bilateral functional neck dissections should be considered in all patients with palpable MTC. PMID- 10363904 TI - Peptide and glycopeptide dendrimers. Part II. AB - Recent progress in peptide and glycopeptide chemistry make the preparation of peptide and glycopeptide dendrimers of acceptable purity, with designed structural and immunochemical properties reliable. New methodologies using unprotected peptide building blocks have been developed to further increase the possibilities of their design and improve their preparation and separation. The sophisticated design of peptide and glycopeptide dendrimers has led to their use as antigens and immunogens, for serodiagnosis and other biochemical uses including drug delivery. Dendrimers bearing peptide with predetermined secondary structures are useful tools in protein de novo design. This article covers synthesis and applications of multiple antigen peptides (MAPs), multiple antigen glycopeptides (MAGs), multiple antigen peptides based on sequential oligopeptide carriers (MAP-SOCs), glycodendrimers and template-assembled synthetic proteins (TASPs). In part II the preparation of MAPs, and the utility of glycodendrimers and TASPs are discussed. PMID- 10363905 TI - Epitope analysis of the multiphosphorylated peptide alpha s1-casein (59-79). AB - The multiphosphorylated tryptic peptide alpha(s1)-casein(59-79) has been shown to be antigenic with anti-casein antibodies. In an approach to determine the amino acyl residues critical for antibody binding we undertook an epitope analysis of the peptide using overlapping synthetic peptides. With alpha(s1)-casein(59-79) as the adsorbed antigen in a competitive ELISA only two of five overlapping synthetic peptides at 1 mM significantly inhibited binding of the anti-casein antibodies. Peptides Glu-Ser(P)-Ile-Ser(P)-Ser(P)-Ser(P)-Glu-Glu and Ile-Val-Pro Asn-Ser(P)-Val-Glu-Glu inhibited antibody binding by 20.0+/-3.6% and 60.3+/-7.9%, respectively. The epitope of Glu63-Ser(P)-Ile-Ser(P)-Ser(P)-Ser(P)-Glu-Glu70 was further localised to the phosphoseryl cluster as the peptide Ser(P)-Ser(P)-Ser(P) significantly inhibited binding of the anti-casein antibodies to alpha(s1) casein(59-79) by 29.5+/-7.4%. Substitution of Ser(P)75 with Ser75 in the second inhibitory peptide Ile-Val-Pro-Asn-Ser(P)75-Val-Glu-Glu also abolished inhibition of antibody binding to x(s1)-casein (59-79) demonstrating that Ser(P)75 is also a critical residue for recognition by the antibodies. These data show that the phosphorylated residues in the cluster sequence -Ser(P)66-Ser(P)-Ser(P)68 and in the sequence -Pro73-Asn-Ser(P)-Val-Glu77- are critical for antibody binding to x(s1)-casein(59-79) and further demonstrate that a highly phosphorylated segment of a protein can be antigenic. PMID- 10363906 TI - (D-(p-benzoylphenylalanine)13, tyrosine19)-melanin-concentrating hormone, a potent analogue for MCH receptor crosslinking. AB - A photoreactive analogue of human melanin-concentrating hormone was designed, [D Bpa13,Tyr19-MCH, containing the D-enantiomer of photolabile p benzoylphenylalanine (Bpa) in position 13 and tyrosine for radioiodination in position 19. The linear peptide was synthesized by the continuous-flow solid phase methodology using Fmoc-strategy and PEG-PS resins, purified to homogeneity and cyclized by iodine oxidation. Radioiodination of [D-Bpa13,Tyr19]-MCH at its Tyr19 residue was carried out enzymatically using solid-phase bound glucose oxidase/lactoperoxidase, followed by purification on a reversed-phase mini-column and HPLC. Saturation binding analysis of [125I]-[D-Bpa13,Tyr19]-MCH with G4F-7 mouse melanoma cells gave a K(D) of 2.2+/-0.2 x 10(-10) mol/l and a B(max) of 1047+/-50 receptors/cell. Competition binding analysis showed that MCH and rANF(1 28) displace [125I]-[D-Bpa13,Tyr19]-MCH from the MCH binding sites on G4F-7 cells whereas alpha-MSH has no effect. Receptor crosslinking by UV-irradiation of G4F-7 cells in the presence of [125I]-[D-Bpa13,Tyr19]-MCH followed by SDS polyacrylamide gel electrophoresis and autoradiography yielded a band of 45-50 kDa. Identical crosslinked bands were also detected in B16-F1 and G4F mouse melanoma cells, in RE and D10 human melanoma cells as well as in COS-7 cells. Weak staining was found in rat PC12 phaeochromocytoma and Chinese hamster ovary cells. No crosslinking was detected in human MP fibroblasts. These data demonstrate that [125I]-[D-Bpa13,Tyr19]-MCH is a versatile photocrosslinking analogue of MCH suitable to identify MCH receptors in different cells and tissues; the MCH receptor in these cells appears to have the size of a G protein coupled receptor, most likely with a varying degree of glycosylation. PMID- 10363907 TI - Nitric oxide and cyclic GMP induce vesicle release at Drosophila neuromuscular junction. AB - Nitric oxide (NO) diffuses as short-lived messenger through the plasma membrane and serves, among many other functions, as an activator of the cGMP synthesizing enzyme soluble guanylyl cyclase (sGC). In view of recent genetic investigations that postulated a retrograde signal from the larval muscle fibers to the presynaptic terminals, we looked for the presence of an NO/cGMP signaling system at the neuromuscular junction (NMJ) of Drosophila melanogaster larvae. Application of NO donors induced cGMP immunoreactivity in the presynaptic terminals but not the postsynaptic muscle fibers at an identified NMJ. The NO induced cGMP immunoreactivity was sensitive to a specific inhibitor (ODQ) of the sGC. Since presynaptic terminals which were surgically isolated from the central nervous system are capable of synthesizing cGMP, we suggest that an NO-sensitive guanylyl cyclase is present in the terminal arborizations. Using a fluorescent dye that is known to stain recycling synaptic vesicles, we demonstrate that NO donors and membrane permeant cGMP analogues cause vesicle release at the NMJ. Moreover, the NO-induced release could be blocked by the specific inhibitor of the sGC. A destaining of synaptic terminals after NO exposure in Ca2+-free solution in the presence of cobalt chloride as a channel blocker suggested that NO stimulates Ca2+-independent vesicle release at the NMJ. The combined immunocytochemical and exocytosis imaging experiments imply the involvement of cGMP and NO in the regulation of vesicle release at the NMJ of Drosophila larvae. PMID- 10363908 TI - Regulation of Drosophila FMRFamide neuropeptide gene expression. AB - Physiologically important peptides are often encoded in precursors that contain several gene products; thus, regulation of expression of polypeptide proteins is crucial to transduction pathways. Differential processing of precursors by cell- or tissue-specific proteolytic enzymes can yield messengers with diverse distributions and dissimilar activities. FMRFamide-related peptides (FaRPs) are present throughout the animal kingdom and affect both neural and gastrointestinal functions. Organisms have several genes encoding numerous FaRPs with a common C terminal structure but different N-terminal amino acid extensions. We have isolated SDNFMRFamide, DPKQDFMRFamide, and TPAEDFMRFamide contained in the Drosophila FMRFamide gene. To investigate the regulation of expression of FMRFamide peptides, we generated antisera to distinguish among the three neuropeptides. We have previously reported the distribution of SDNFMRFamide and DPKQDFMRFamide. In this article, we describe TPAEDFMRFamide expression. TPAEDFMRFamide antisera stain cells in embryonic, larval, pupal, and adult thoracic and abdominal ganglia. In addition, TPAEDFMRFamide-immunoreactive material is present in a lateral protocerebrum cell in adult. Thus, TPAEDFMRFamide antisera staining of neural tissue is different from SDNFMRFamide or DPKQDFMRFamide. In addition, TPAEDFMRFamide antisera stain larval, pupal, and adult gut, while SDNFMRFamide and DPKQDFMRFamide do not. TPAEDFMRFamide immunoreactivity is present in cells stained by FMRFamide antisera. Taken together, these data support the conclusion that TPAEDFMRFamide is differentially processed from the FMRFamide polypeptide protein precursor and may act in both neural and gastrointestinal tissue. PMID- 10363909 TI - Androgen receptor immunoreactivity in the male and female Syrian hamster brain. AB - To investigate potential mechanisms for sex differences in the physiologic response to androgens, the present study compared the hormonal regulation of intracellular androgen receptor partitioning and the distribution of androgen receptor immunoreactivity in select brain regions from male and female hamsters. Androgen receptors were visualized on coronal brain sections. Two weeks after castration, androgen receptor immunoreactivity filled the neuronal nuclei and cytoplasm in males and females. In gonad-intact males and females, androgen receptor immunoreactivity was limited to the cell nucleus. Whereas exogenous dihydrotestosterone prevented cytoplasmic immunoreactivity, estrogen at physiologic levels did not. These results suggest that nuclear androgen receptor immunoreactivity in gonad-intact females is maintained by endogenous androgens, and that androgens have the potential to influence neuronal activity in either sex. However, sex differences in the number and staining intensity of androgen responsive neurons were apparent in select brain regions. In the ventral premammillary nucleus, ventromedial nucleus of the hypothalamus, and medial amygdaloid nucleus, androgen receptor staining was similar in gonadectomized males and females. In the lateral septum, posteromedial bed nucleus of the stria terminalis (BNSTpm), and medial preoptic nucleus, the number of androgen receptor immunoreactive neurons was significantly lower in females (p < .05). Moreover, the integrated optical density/cell in BNSTpm was significantly less in females (1.28+/-0.3 units) than in males (2.21+/-0.2 units; p < .05). These sex differences in the number and staining intensity of androgen-responsive neurons may contribute to sex differences in the behavioral and neuroendocrine responses to androgens. PMID- 10363911 TI - Developmental expression of P-glycoprotein (multidrug resistance gene product) in the rat brain. AB - P-Glycoprotein (PGP), a product of the multidrug resistance gene (mdr), acts as an adenosine triphosphate-dependent drug efflux system in cells. Initially, PGP was found in cancer cells, but it is now known that PGP is richly distributed in the adult brain. Passage to the central nervous system is limited by the blood brain barrier (BBB), and mdr1 gene-deficient mice showed up-regulation of BBB permeability. In this study, we examined the expression and localization of PGP in the rat brain during development. PGP protein was predominantly detected in the membrane fraction of the adult rat brain, although it was also faintly detected in the cytosolic fraction. PGP protein in the membrane fraction was undetectable in the embryo and early stages of postnatal development by immunoblotting studies, was first detected on postnatal day (P) 7, and then gradually increased to reach a plateau. Such changes were observed commonly in the cerebral cortex, hippocampus, and cerebellum. Immunohistochemical studies showed that PGP immunoreactivity was first detected on P7, and intense PGP immunoreactivity was observed in the adult rat brain. Double-immunolabeling studies revealed that PGP was colocalized with von Willebrand factor immunoreactive capillaries. We further examined the colocalization of PGP and astrocytes using glial fibrillary acidic protein (GFAP) as a marker. Three dimensional analysis showed that the GFAP-immunoreactive astrocytes possessed fine processes which ensheathed capillaries, but the PGP immunoreactivity did not colocalize with the GFAP immunoreactivity. These results indicate that PGP expression increased with postnatal development and is localized in the brain capillaries. PMID- 10363910 TI - Beta-sheet breaker peptide inhibitor of Alzheimer's amyloidogenesis with increased blood-brain barrier permeability and resistance to proteolytic degradation in plasma. AB - Short synthetic peptides homologous to the central region of Abeta but bearing proline residues as beta-sheet blockers have been shown in vitro to bind to Abeta with high affinity, partially inhibit Abeta fibrillogenesis, and redissolve preformed fibrils. While short peptides have been used extensively as therapeutic drugs in medicine, two important problems associated with their use in central nervous system diseases have to be addressed: (a) rapid proteolytic degradation in plasma, and (b) poor blood-brain barrier (BBB) permeability. Recently, we have demonstrated that the covalent modification of proteins with the naturally occurring polyamines significantly increases their permeability at the BBB. We have extended this technology to iAbeta11, an 11-residue beta-sheet breaker peptide that inhibits Abeta fibrillogenesis, by covalently modifying this peptide with the polyamine, putrescine (PUT), and evaluating its plasma pharmacokinetics and BBB permeability. After a single intravenous bolus injection in rats, both 125I-YiAbeta11 and 125I-PUT-YiAbeta11 showed rapid degradation in plasma as determined by trichloroacetic acid (TCA) precipitation and paper chromatography. By switching to the all D-enantiomers of YiAbeta11 and PUT-YiAbeta11, significant protection from degradation by proteases in rat plasma was obtained with only 1.9% and 5.7% degradation at 15 min after intravenous bolus injection, respectively. The permeability coefficient x surface area product at the BBB was five- sevenfold higher in the cortex and hippocampus for the 125I-PUT-D-YiAbeta11 compared to the 125I-D-YiAbeta11, with no significant difference in the residual plasma volume. In vitro assays showed that PUT-D-YiAbeta11 retains its ability to partially inhibit Abeta fibrillogenesis and dissolve preformed amyloid fibrils. Because of its five- to sevenfold increase in permeability at the BBB and its resistance to proteolysis in the plasma, this polyamine-modified beta-sheet breaker peptide may prove to be an effective inhibitor of amyloidogenesis in vivo and, hence, an important therapy for Alzheimer's disease. PMID- 10363913 TI - Dibutyryl cyclic AMP-induced process formation in astrocytes is associated with a decrease in tyrosine phosphorylation of focal adhesion kinase and paxillin. AB - Focal adhesion kinase (FAK or pp125FAK) is a cytosolic protein tyrosine kinase which plays an important role in integrin-mediated signal transduction. Adhesion of cells to the substratum correlates with an increase in tyrosine phosphorylation of FAK as well as an associated protein, paxillin. In this report we show that the tyrosine phosphorylation of FAK and paxillin are decreased during dibutyryl cyclic AMP-induced (dB-cAMP) process formation in astrocytes. When astrocytes in suspension are treated with dB-cAMP, no alteration in morphology or tyrosine phosphorylation is observed, suggesting that both phenomena are linked and adhesion dependent. Furthermore, genistein, a tyrosine kinase inhibitor, can induce process formation in such cells, underscoring the significance of protein tyrosine kinases in maintaining the morphology of adherent cells. Finally, endothelin-1, a vasopeptide which is known to inhibit process formation in astrocytes, inhibited the tyrosine dephosphorylation of proteins associated with dB-cAMP treatment. These results suggest that the formation of asymmetric processes in astrocytes results from a coordinated set of alterations in the actin cytoskeleton as well as the adhesion of the cell to the substratum. Modification of the properties of such molecules is required for process formation and the dynamic modulation of astrocytic morphology in vitro and in vivo. PMID- 10363912 TI - Cortical neurite outgrowth and growth cone behaviors reveal developmentally regulated cues in spinal cord membranes. AB - Corticospinal axon outgrowth in vivo and the ability to sprout or regenerate after injury decline with age. This developmental decline in growth potential has been correlated with an increase in inhibitory myelin-associated proteins in older spinal cord. However, previous results have shown that sprouting of corticospinal fibers after contralateral lesions begins to diminish prior to myelination, suggesting that a decrease in growth promoting and/or an increase in inhibitory molecules in spinal gray matter may also regulate corticospinal axon outgrowth. To address this possibility, we carried out in vitro experiments to measure neurite outgrowth from explants of 1-day-old hamster forelimb sensorimotor cortex that were plated onto membrane carpets or membrane stripe assays prepared from white or gray matter of 1-to 22-day-old cervical spinal cord. On uniform carpets and in the stripe assays cortical neurites grew robustly on young but not older membranes from both white and gray matter. Mixtures of membranes from 1- and 15-day spinal cord inhibited neurite outgrowth, suggesting that the presence of inhibitory molecules in the 15-day cord overwhelmed permissive or growth promoting molecules in membranes from 1-day cord. Video microscopic observations of growth cone behaviors on membrane stripe assays transferred to glass coverslips supported this view. Cortical growth cones repeatedly collapsed at borders between permissive substrates (laminin or young membrane stripes) and nonpermissive substrates (older membrane stripes). Growth cones either turned away from the older membranes or reduced their growth rates. These results suggest that molecules in both the gray and white matter of the developing spinal cord can inhibit cortical neurite outgrowth. PMID- 10363915 TI - Estrogen receptor expression in the facial nucleus of adult hamsters: does axotomy recapitulate development? AB - Testosterone propionate (TP) augments hamster facial motoneuron regeneration following axonal injury by an androgen-mediated mechanism. Although many of the trophic properties of TP are androgenic, TP can be metabolized to estradiol (E). We have recently shown that E administered in supraphysiological doses can also enhance facial nerve regeneration. The mechanism by which E alters nerve regeneration is unknown. The recent discovery of transient estrogen receptor (ER) expression in the developing rat facial motor nucleus (FMN), coupled with the concept that regeneration may recapitulate development, has led to the hypothesis that facial nerve injury may transiently induce expression of ER in the adult hamster FMN or one of its chief afferents, the principal nucleus of the trigeminal nerve (Nu5). In the present study, this hypothesis was tested using steroid hormone autoradiographic procedures. The right facial nerve was injured in castrated or castrated plus TP adult hamsters. A gonadally intact, nonaxtomized group of hamsters was also included to examine constitutive expression of ER in the FMN or Nu5. The paraventricular nucleus of the hypothalamus (PVN; positive control), FMN, and Nu5, were qualitatively and quantitatively examined for the presence of ER. As expected, ER were present in the PVN-positive control in all groups. ER were neither present nor induced with facial nerve injury or TP administration in either the FMN or Nu5. Alternate mechanisms by which E enhancement of facial nerve regeneration without ER might be explained are discussed. PMID- 10363914 TI - A shift in protein S-palmitoylation, with persistence of growth-associated substrates, marks a critical period for synaptic plasticity in developing brain. AB - In the mammalian cortex, the initial formation of synaptic connections is followed by a prolonged period during which synaptic circuits are functional, but retain an elevated capacity for activity-dependent remodeling and functional plasticity. During this period, synaptic terminals appear fully mature, morphologically and physiologically. We show here, however, that synaptic terminals during this period are distinguished by their simultaneous accumulation of multiple growth-associated proteins at levels characteristic of axonal growth cones, and proteins involved in synaptic transmitter release at levels characteristic of adult synapses. We show further that newly formed synapses undergo a switch in the dynamic S-palmitoylation of proteins early in the critical period, which includes a large and specific decrease in the palmitoylation of GAP-43 and other major substrates characteristic of growth cones. Previous studies have shown that a similar reduction in ongoing palmitoylation of growth cone proteins is sufficient to stop advancing axons in vitro, suggesting that a developmental switch in protein S-palmitoylation serves to disengage the molecular machinery for axon extension in the absence of local triggers for remodeling during the critical period. Only much later does a decline in the availability of major growth cone components mark the molecular maturation of cortical synapses at the close of the critical period. PMID- 10363916 TI - The gene mod(mdg4) affects synapse specificity and structure in Drosophila. AB - The mechanisms by which synapse assembly and maturation are orchestrated during development are largely unknown. We used P-element mutagenesis and a larval anatomical screen to isolate mutants in which synapse structure was altered. Here, we describe a mutation isolated with this screen, branch point disrupted (bpd), in which both synapse specificity and synapse morphology were altered. Synaptic terminals in bpd mutants developed abnormally, forming multiple branch points, overgrowing to inappropriate neighboring muscles, and establishing aberrant folding of postsynaptic membranes. Ultrastructural characterization of synaptic boutons in bpd demonstrated abnormal layering of the postsynaptic specialization or subsynaptic reticulum (SSR). Genetic and molecular analyses revealed that bpd is an allele of mod(mdg4), a gene coding for a protein with many similarities to transcription factors, which has been implicated in the regulation of chromatin insulation. Our results suggest that mod(mdg4) may regulate a gene(s) essential to normal synapse formation. PMID- 10363917 TI - The voltage gated potassium channel KCNQ2 and idiopathic generalized epilepsy. AB - Mutations in the voltage gated potassium channel gene KCNQ2 and the homologous gene KCNQ3 have been found to cause a rare monogenic subtype of idiopathic generalized epilepsy, the benign familial neonatal convulsions. Recently, the heteromeric KCNQ2/KCNQ3 channel was found to contribute to the native M-current, one of the most important regulators of neuronal excitability. By performing a systematic mutation scan of the coding region and an association study involving a frequent Thr752Asn substitution polymorphism, we, therefore, investigated whether allelic variation of the KCNQ2 gene confers susceptibility to common subtypes of idiopathic generalized epilepsy. Our results do not provide evidence that allelic variation of the KCNQ2 gene contributes a common and relevant effect to the pathogenesis of common subtypes of idiopathic generalized epilepsy. PMID- 10363918 TI - Attention changes the peak latency of the visual gamma-band oscillation of the EEG. AB - To investigate the physiological role of visual gamma-band oscillation (GBO), we calculated the event-related dynamics of the EEG power-spectrum for paired visual stimuli (S1 and S2) with or without attention in 12 subjects. The visual stimuli elicited transient increases in the GBO power (around 40 Hz), which were maximal over the parietal area. The peak GBO increase appeared around 300 ms after stimulus onset, but its latency was shorter after S1 and longer after S2 under the 'with attention' than under the 'without attention' condition. This transient increase in the visual GBO is thought to reflect attention and to reset the activity of the visual system in preparation for a new stimulus. PMID- 10363919 TI - Nociceptin system plays a role in the memory retention: involvement of naloxone benzoylhydrazone binding sites. AB - We investigated the role of nociceptin system in learning and memory in mice. The deficiency of nociceptin receptors and nociceptin itself did not affect the alternation behavior in the Y-maze test. In the passive avoidance test, the step through latencies of nociceptin receptor knockout mice were longer than those of wild-type mice. Nociceptin shortened the step-through latency in wild-type mice. This impairment on passive avoidance task was reversed by naloxone benzoylhydrazone (NalBzoH), indicating that the amnesic effects of nociceptin may be mediated through the NalBzoH recognition sites. These suggest that nociceptin system plays an important role in the memory retention of passive avoidance task, and NalBzoH-recognized sites are involved in nociceptin-induced impairment of the memory retention. PMID- 10363920 TI - Labelling of rat vagal preganglionic neurones by carbocyanine dye DiI applied to the heart. AB - We describe a method for applying the carbocyanine dye DiI to the rat heart that takes advantage of the dye's lipophilic properties and its ability to diffuse easily into tissues, and results in specific retrograde labelling of cardiac vagal preganglionic neurones in the medulla oblongata. Most of the labelled neurones were found bilaterally in the nucleus ambiguus (81%), with a few sparsely distributed in the dorsal motor vagal nucleus (6.5%), and in an intermediate area located between these two nuclei (12.5%). We contend that the method of applying DiI crystals to the surface of the heart is a more efficient, accurate and reproducible method of retrograde labelling than the injection of tracers into this very delicate tissue. PMID- 10363921 TI - The arcuate nucleus is pivotal in mediating the anorectic effects of centrally administered leptin. AB - The adipose tissue hormone leptin, which is secreted to the general circulation and transported into the brain in a facilitated manner, possibly acts via hypothalamic neurones to reduce food intake and increase energy expenditure. To evaluate the involvement of importance of the arcuate nucleus in leptin induced anorexia, groups of rats treated neonatally with monosodium-glutamate (MSG; arcuate lesioned) and littermate controls were injected centrally with 5 microg recombinant leptin or saline daily for three consecutive days. Leptin significantly inhibited food intake and caused weight-loss in non-MSG rats ( 14.5+/-3.0 g vs. 10.2+/-4.3 g; mean +/-s.e.m.; leptin vs. vehicle) whereas MSG treated rats were unresponsive to leptin treatment (5.0+/-2.2 g vs. 0.8+/-3.8 g; leptin vs. vehicle). The present data indicate that an intact arcuate nucleus is necessary for leptins actions on food intake and body weight. PMID- 10363922 TI - The bacterial Neo gene confers neomycin resistance to mammalian cochlear hair cells. AB - The aminoglycoside antibiotics are important agents in the treatment of bacterial infection. At pharmacological doses they also preferentially damage the sensory hair cells of the inner ear, leading to ototoxic hearing loss. However, it has been suggested that the mechanism of ototoxicity is different from that of bacterial toxicity. The bacterial neomycin phosphotransferase gene (Neo) confers resistance to this and related aminoglycoside antibiotics. To determine whether the Neo gene also confers resistance to vertebrate ototoxicity, the sensitivity of cochlear hair cells to neomycin was evaluated in mice with a targeted insertion of Neo. Organotypic cultures of the organ of Corti, isolated from neonatal wild-type mice and two strains of mice carrying the Neo gene, were cultured for 72 h in the absence (controls) or in the presence of 200 microM neomycin. Organs from wild-type mice showed no remaining outer hair cells and <25% of inner hair cells when incubated with neomycin. In contrast, organs from mice carrying the Neo gene showed no loss of hair cells after neomycin treatment. PMID- 10363923 TI - Alteration of spinal cord IGF-I receptors and skeletal muscle IGF-I after hind limb immobilization in the rat. AB - The effects of 4 weeks' hind-limb immobilization on the spinal cord insulin-like growth factor-I (IGF-I) receptors and skeletal muscle IGF-I level was investigated in rats. Quantitative receptor autoradiography using [125I]IGF-I as a ligand was performed to measure IGF-I receptors in cryosections from the lumbar region of the spinal cord. IGF-I receptor levels were significantly higher in all spinal cord laminae on the side ipsilateral to the immobilized limb than in the same spinal level of the controls. Using radioimmunoassay (RIA), IGF-I levels were significantly low in the soleus (SOL), but not the tibialis anterior (TIB) muscles, compared to the controls. The enhancement of the spinal cord IGF-I receptors after hind-limb immobilization may constitute part of the nervous system response to disuse. PMID- 10363924 TI - Basic fibroblast growth factor enhances axonal sprouting after cortical injury in rats. AB - The trophic factors responsible for initiating and guiding the outgrowth of axons have proven to be elusive throughout most of this century. Entorhinal cortex injury, which denervates the hippocampal formation of rats, induces axonal sprouting by several surviving hippocampal afferents and results in a significant elevation of growth factors, one of which is basic fibroblast growth factor (bFGF). The possibility that bFGF may regulate lesion-induced hippocampal sprouting was examined by making i.v. bFGF infusions into rats with unilateral entorhinal lesions. Basic FGF treatment significantly increased sprouting by the cholinergic septodentate pathway. Thus, the increase in bFGF following central nervous system injury may signal its role in the regulation of injury-related axonal remodeling of a cholinergic pathway. PMID- 10363925 TI - Regulation of inner ear fluid in the rat by vasopressin. AB - The anti-diuretic hormone vasopressin has been shown to be important in regulating inner ear fluid. The diuretic hormone, CNP, and its receptor, ANP-B receptor, may also function in the regulation of inner ear fluid. To determine whether vasopressin directly affects the fluid level, we infused this hormone to rat and assay of V2-AVP receptor mRNA by semiquantitative RT-PCR demonstrated a significantly lower level of this transcript in vasopressin-infused animals than in saline-infused animals. The levels of CNP and ANP-B receptors mRNA, however, were the same in both groups of rats. Results suggest that high plasma levels of vasopressin may be a principal causal factor of endolymphatic hydrops in Meniere's disease, perhaps by down-regulating the number of vasopressin receptors. PMID- 10363926 TI - A role for nitric oxide in the median eminence and arcuate nucleus response to capsaicin treatment in rats. AB - This study examined a possible functional involvement of nitric oxide (NO) in the median eminence (ME) and arcuate nucleus (ARC) after capsaicin treatment in rats. Subcutaneous injection of capsaicin increased nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) activity in the ARC-ME compared with vehicle treatment. Fos expression was increased in the ARC after capsaicin injection compared with vehicle-treated rats. Pretreatment with the NO synthase (NOS) inhibitor, N(omega)-nitro-L-arginine methyl ester (L-NAME) attenuated the effect of capsaicin on Fos expression and NADPH-d reactivity in the ARC-ME in comparison with rats injected with D-NAME, the inactive stereoisomer of L-NAME. These observations suggest that NO makes a major contribution to the response of the ARC-ME to a stressor such as capsaicin. PMID- 10363927 TI - Delayed 5-HT release in the developing cortex of microencephalic rats. AB - Postnatal changes in the fiber distribution and release of 5-HT in the somatosensory cortex (Sm) of methylazoxymethanol acetate (MAM)-induced microencephalic rats were studied. A transient accumulation of serotonergic fibers was observed in the Sm of the control and MAM rats in the first 2 weeks following birth. However, the density of serotonergic fibers was higher in the MAM rats than in the controls, and the distribution pattern of serotonergic fibers was more extensive in the MAM rats. The microdialysis indicated a high basal level and a delayed K+ -evoked 5-HT release in the Sm of MAM rats on postnatal day 10. The present results suggest morphological and functional alterations of serotonergic fibers in the Sm of MAM rats during the critical period of cortical formation. PMID- 10363928 TI - Galanin expression is decreased by cAMP-elevating agents in cultured sympathetic ganglia. AB - Galanin expression is co-regulated in peripheral neurons with that of vasoactive intestinal peptide (VIP) under a variety of conditions. For example, the expression of both increase after explantation of adult rat superior cervical ganglia (SCG). Because VIP participates in a positive feedback loop regulating its own expression, we examined whether VIP also increases galanin expression. Galanin mRNA and peptide are nearly undetectable in the SCG in vivo, but increase dramatically after 24-48 h in organ culture. Addition of VIP or forskolin to the culture medium reduced galanin mRNA expression by 75% and 77%, respectively, and reduced galanin peptide expression by 76% and 82%, respectively, compared with ganglia cultured in control medium. In contrast, isoproterenol stimulation did not significantly alter levels of galanin mRNA or peptide, consistent with previous observations that isoproterenol exerts its effect on SCG non-neuronal cells, but not on neurons. The results indicate that galanin and VIP are differentially regulated in sympathetic neurons by cAMP- elevating agents. PMID- 10363929 TI - Modulatory action of noradrenergic system on spinal motoneurons in humans. AB - Previous findings in animals demonstrated that the noradrenergic coeruleospinal system exerts a tonic facilitation on spinal reflexes and that activation of alpha2-autoinhibitory receptors can be responsible for a disfacilitation of the spinal activity. To investigate this issue further, we examined whether this system is also involved in descending facilitatory control of spinal motoneurons in healthy humans. The H-reflex technique was utilized to assay the motoneuronal excitability. The ratio between the maximal reflex response (H) and maximal direct response (M) was determined in each subject and was calculated at 10 min intervals before and after i.v. administration of the alpha2-agonist clonidine (0.5 microg/kg). In all subjects a marked decrease of the H/M ratio, due to depression of the H response, occurred 10 min following the clonidine injection and reached its maximum within 30 min. No significant changes of blood pressure values were provoked by drug injections. These results suggest that an autoinhibitory action may be induced by alpha2-receptor activation of locus coeruleus neurons in humans, and that this device may serve as a mechanism for a myotonolytic action on spinal motoneurons. PMID- 10363930 TI - Serotonin-6 receptor variant (C267T) and clinical response to clozapine. AB - Clozapine is an effective atypical antipsychotic that has high affinity for serotonin type 6 receptors (5HT6). We tested the hypothesis that clinical response to clozapine in patients refractory to typical antipsychotic treatment is related to the genetic variant (C267T) of the 5HT6 receptors. Ninety-nine schizophrenic patients with a history of non-response to typical antipsychotics were included in the study. The results demonstrated a modest but significant relationship between presence of the variant of the 5HT6 receptors and the response to clozapine in these patients. Patients with homogenous 267T/T genotype had a better response than other patients. Although replication is required, these results suggest that the 5HT6 receptor C267T polymorphism may be involved in clozapine response, especially in patients with anxious or depressed symptoms. PMID- 10363931 TI - Familial influences on cortical evoked potentials in migraine. AB - Cortical information processing in migraine patients is impaired between attacks, showing deficient habituation of pattern-reversal visual evoked potentials (VEP), and strong intensity dependence of auditory cortical evoked potentials (IDAP). This could be a genetic trait as certain genetic patterns are known for evoked potentials in healthy subjects. We investigated VEP habituation and IDAP in 20 pairs of migraineurs made up of parents and their children. Using a Monte-Carlo statistical method, we selectively assessed vertical familial influences. VEP habituation and IDAP were abnormal in both parents and children. However, similarity was far more pronounced between related pairs than between unrelated pairs. Familial influences are highly significant in determinants of cortical information processing in migraineurs, hence supporting the important role of genetic factors. PMID- 10363932 TI - CBP associates with the p42/p44 MAPK enzymes and is phosphorylated following NGF treatment. AB - The ability of the CBP (CREB binding protein) coactivator to stimulate transcription has previously been shown to be stimulated by treatment of neuronal cells with nerve growth factor (NGF). This effect is dependent upon activation of the p42/p44 MAPK (mitogen activated protein kinase) pathway. Here we show that both CBP and the related p300 protein directly associate with the p42/p44 MAPK enzymes both prior to and following their activation by NGF and that CBP is phosphorylated following NGF treatment. These results indicate that phosphorylation of CBP itself by the p42/p44 MAPK pathway is likely to be critical for its role in NGF-mediated stimulation of gene expression. PMID- 10363933 TI - Differential inhibition of chromatic and achromatic perception by transcranial magnetic stimulation of the human visual cortex. AB - The magnocellular visual pathway is devoted to low-contrast achromatic and motion perception whereas the parvocellular pathway deals with chromatic and high resolution spatial vision. To specifically separate perception mediated by these pathways we have used low-contrast Gaussian filtered black-white or coloured visual stimuli. By use of transcranial magnetic stimulation (TMS) over the visual cortex inhibition of magnocellular stimuli was achieved distinctly earlier by about 40 ms compared with parvocellular information. A nonspecific inhibition of all stimuli could be seen peaking at 75-90 ms, significantly higher for magnocellular stimuli. The particular vulnerability of magnocellular stimuli to TMS is correlated with distinct physiological properties of this pathway such as faster conduction velocity and non-linear stimulus encoding. PMID- 10363934 TI - DNA methylation at the putative promoter region of the human dopamine D2 receptor gene. AB - DNA methylation was investigated in the putative promoter region of the human dopamine D2 receptor gene (DRD2). Twenty-two DNA samples from two types of cells differentially expressing D2 receptors, striatum and lymphocytes, were subjected to bisulphite modification-based mapping of methylated cytosines. In the tested region, the DNA from lymphocytes exhibited a significantly higher degree of methylation than that from striata. In addition, a significantly higher proportion of methylated to unmethylated cytosines was detected in DRD2 from the right than the left striatum, and a trend towards a greater degree of methylation was detected in older than in younger individuals. These DRD2 methylation findings are consistent with dopamine D2 receptor binding data from the literature which support the idea that DNA methylation plays a role in regulation of DRD2 expression. PMID- 10363936 TI - Delayed implantation of nigral grafts improves survival of dopamine neurones and rate of functional recovery. AB - In order to test the hypothesis that poor survival of dopaminergic neurones in nigral transplants may be due, at least in part, to acute toxic changes in the host striatum within the first hour after injury, we experimentally evaluated the consequences of imposing a brief delay (20 min, 1 or 3 h) between positioning the injection cannula and extruding the graft tissue. A delay of as little as 1 h resulted in a three-fold increase in survival of dopamine neurones in the grafts and a more rapid abolition of amphetamine-induced rotational asymmetry in the host animals. These results suggest that acute but rapidly resolving changes in the host striatal environment induced by the implantation procedure itself can have a significantly deleterious effect on the survival of embryonic nigral grafts. PMID- 10363935 TI - Mitochondrial participation in modulation of calcium transients in DRG neurons. AB - The effects of the mitochondrial Na+/Ca2+ exchange blocker tetraphenylphosphonium (TPP+) and the permeability transition blocker cyclosporinA (CysA) on the ability of mitochondria to participate in the regulation of intracellular calcium were investigated on freshly isolated mice sensory DRG neurons. The free intracellular calcium level ([Ca2+]in) was measured using indo-1 based microfluorimetry. The characteristics of depolarization-induced [Ca2+]in transients were changed in the presence of 25 microM TPP+. The amplitude of [Ca2+]in transients became decreased and the restoration of resting [Ca2+]in level speeded up in the presence of TPP+. Application of 5 microM cyclosporinA induced substantial residual elevation of [Ca2+]in after termination of depolarization. We conclude that the mitochondrial Na+/Ca2+ exchanger mechanism plays an important role in the regulation of calcium signals during neuronal activity, prolonging them by releasing Ca2+ stored during transient peak. Activation of permeability transition pores does not participate in these processes. PMID- 10363937 TI - Synergistic effects of laminin and thyroid hormones on neuron polarity in culture. AB - This study is aimed to elucidate whether triiodothyronine (T3) and laminin have additive effects in regulating neural differentiation. We focused our attention on the expression of synapsin I, the 68 kDa component of the neurofilament triplet (NF-68), growth associated protein (GAP)-43 and microtubule associated protein (MAP)-2 as markers of synapses, cytoskeleton, axons and the somatodendritic domain, respectively. The addition of T3 to the medium of differentiating rat cortical neurons cultured on laminin did not have any effect on the concentration of these proteins, but was critical for their subcellular localization, suggesting a synergistic role of thyroid hormones and laminin in the establishment of neural polarity. PMID- 10363938 TI - Diazepam induces FGF-2 and its mRNA in the spinal cord. AB - We have explored the possibility that the GABA system can influence the expression of FGF-2 in the spinal cord. The GABA agonist diazepam was systemically injected in adult rats, and the expression of FGF-2 was examined in the cervical spinal cord region between 6 h and 7 days post-injection. Results of nuclease protection assays showed increases in FGF-2 mRNA, starting by 6 h and returning to approximately control values by 3 days. There was an increase in the density of FGF-2 immunostained astrocyte-like cells between 6 h and 3 days. Results showing that diazepam up-regulates FGF-2 expression in the spinal cord suggest that GABA may promote neuroplasticity in concert with FGF-2. PMID- 10363939 TI - Does stimulus quality affect the physiologic MRI responses to brief visual activation? AB - We studied the effect of stimulus quality on the basic physiological response characteristics of oxygenation-sensitive MRI signals. Paradigms comprised a contrast-reversing checkerboard vs. darkness or vs. gray light as well as gray light vs. darkness in a 2 s/52 s protocol (nine subjects). MRI was performed at 2.0 T using single-shot gradient-echo EPI (TR/TE = 500/54 ms, flip angle 30 degrees). All paradigms elicited almost identical signal intensity time courses comprising a latency period (1-2s), an activation-induced signal increase (4-4.5% at about 6-7 s after stimulus onset) and a post-stimulus signal undershoot (-1%) that slowly recovered to baseline (about 50 s). Thus, in contrast to findings for sustained stimulation, brief presentations of distinct visual stimuli exhibit similar physiological response characteristics that support the use of a uniform response profile for the evaluation of event related paradigms. PMID- 10363940 TI - Impaired visual search in dyslexia relates to the role of the magnocellular pathway in attention. AB - We tested the hypothesis that in a cluttered visual scene, the magnocellular (M) pathway is crucial for focusing attention serially on the objects in the field. Since developmental dyslexia is commonly associated with an M pathway deficit, we compared reading impaired children and age-matched normal readers in a search task that required the detection of a target defined by the conjunction of two features, namely form and colour, that are processed by the parvocellular dominated ventral neocortical stream. The dyslexic group's performance was significantly poorer than the controls when there were a large number of distractor items. The scheme of selective attention proposed from these results provides a neural mechanism that underlies reading and explains the pathophysiology of dyslexia. PMID- 10363941 TI - Targeting of marrow-derived astrocytes to the ischemic brain. AB - Bone marrow progenitor cells have been shown to contribute to a small proportion of cells in nonhematopoietic tissues including the brain. In the acute unilateral middle cerebral artery occlusion model in spontaneously hypertensive rats following male-to-female bone marrow transplantation, we present data suggesting that 55% more marrow-derived cells, in general, and 161% more GFAP-positive astrocytes, in particular, migrate preferentially to the ischemic cortex than to the contralateral non-ischemic hemisphere. In addition to their biological significance, our findings could have therapeutic implications. Marrow-derived progenitor cells could potentially be used as vehicles for ex vivo gene transfer to the brain. PMID- 10363942 TI - Expression of p75 neurotrophin receptor in the injured and regenerating rat retina. AB - The low affinity neurotrophin receptor (p75) has been suggested to be involved in apoptosis after neuronal injury. The tempo-spatial expression of p75 in the axotomized and regenerating retinas has not yet been determined. We examined the expression of p75 in the RGCs of these retinas using 192-IgG immunohistochemistry. The failure to detect p75 immunoreactivity on retrogradely fluorogold (FG)-labeled retinal ganglion cells (RGCs) in axotomized and regenerating adult retinas indicated that p75 was not expressed on RGCs. The low affinity p75 receptor was mainly localized on the Muller cell processes identified with vimentin antibody. Since p75 is not present in RGCs, the proposed pro-apoptotic role of p75 involved in the RGC death after optic nerve injury is unlikely to occur in rats. PMID- 10363943 TI - Electrophysiological evidence of two different types of error in the Wisconsin Card Sorting Test. AB - The specificity of the Wisconsin Card Sorting Test (WCST) for assessing frontal lobe pathology remains controversial, although lesion and cerebral blood flow studies continue to suggest a role for the dorsolateral prefrontal cortex in WCST performance. Inconsistencies might derive from the extended use of various WCST scores as equivalent indicators of frontal pathology. In this study, event related potentials (ERPs) were recorded from 32 normal subjects who committed perseverative and non-perseverative errors. Both types of WCST errors evoked anomalous but distinct ERP patterns over frontal lobe regions. Perseverative errors were also associated with a dysfunctional extrastriate response to stimulation. This evidence suggests that perseverative and non-perseverative errors result from disruptions in two different prefrontal neural networks engaged during card sorting. PMID- 10363944 TI - Temporal span of human echoic memory and mismatch negativity: revisited. AB - The stimulus onset asynchrony (SOA)-related decrease in mismatch negativity (MMN) amplitude has been used to infer a putative auditory sensory memory duration of 4 10 s. However, both increased standard-to-standard (SSA) and standard-to-deviant (SDA) gaps could contribute to the effect. Fourteen subjects were presented with standard and deviant tones with short (0.35 s) and long (3.5 s) SOAs. In addition, the SSA and SDA were separately manipulated to test the relative contributions of slower rate of standard tone presentation and longer SDA gap to the SOA-related decrease in MMN amplitude. The MMN amplitude decreased with long SOA by 61%. Increases in SSA and SDA resulted in intermediate 47% and 31% decreases, these manipulations explaining 67% of the long SOA effect (p<0.001). Consequently, echoic memory length cannot be directly inferred from an MMN-SOA dependency function. PMID- 10363945 TI - Superior pre-attentive auditory processing in musicians. AB - The present study focuses on influences of long-term experience on auditory processing, providing the first evidence for pre-attentively superior auditory processing in musicians. This was revealed by the brain's automatic change detection response, which is reflected electrically as the mismatch negativity (MMN) and generated by the operation of sensoric (echoic) memory, the earliest cognitive memory system. Major chords and single tones were presented to both professional violinists and non-musicians under ignore and attend conditions. Slightly impure chords, presented among perfect major chords elicited a distinct MMN in professional musicians, but not in non-musicians. This demonstrates that compared to non-musicians, musicians are superior in pre-attentively extracting more information out of musically relevant stimuli. Since effects of long-term experience on pre-attentive auditory processing have so far been reported for language-specific phonemes only, results indicate that sensory memory mechanisms can be modulated by training on a more general level. PMID- 10363946 TI - Activity of prefrontal neurons during location and color delayed matching tasks. AB - Many cells in prefrontal cortex show enhanced activity prior to movement onset in delayed or memory-guided saccade tasks. This activity is a possible neural correlate of spatial attention and working memory. The goal of this study was to determine whether delay activity is evoked when non-spatial cues such as color are used to guide saccades. Monkeys were trained on a saccade target selection task in which they were cued for either the location or color of the rewarded target. When the location of the target was specified explicitly, many cells showed visual responses and delay activity that were spatially selective. Color selective visual responses or delay activity were both rare and weak. However, for many cells, spatially selective delay activity could be evoked when color was used to specify the location of the target. These results indicate that color is capable of eliciting spatially selective activity from cells that have no overt color selectivity. PMID- 10363947 TI - Protein kinases A and C are opponents in modulating glial Ca2+ -activated K+ channels. AB - The modulation of the activity of Ca2+ -activated K+ (BK) channels by activators of protein kinases A and C, respectively, was studied in cell-attached patches on isolated Muller (retinal glial) cells from rabbits. The BK channel activity was stimulated by membrane depolarization and by increasing of the intracellular Ca2+ concentration. Extracellular exposure to dibutyryl-cAMP, known to stimulate the protein kinase A, increased the open probability of the channels. Exposure to a phorbol ester, as an activator of protein kinase C, strongly reduced the channel activity whereas exposure to the protein kinase inhibitor, staurosporine, stimulated the channel activity. As glial BK channels are modulated in an opposite manner by protein kinases A and C, they may act as a cellular focus of integration of the inputs from different signaling pathways. PMID- 10363949 TI - Involvement of motor cortices in retrieval of kanji studied by functional MRI. AB - Functional magnetic resonance imaging was successfully used to study the activation of the motor cortices during retrieval of Japanese ideogram, kanji. The subjects performed kanji completion tasks to generate a kanji in response to an element which is always written first. In most of the subjects, the contralateral premotor cortex, the presupplementary motor area (pre-SMA) and the bilateral intraparietal sulcus were activated during retrieval of kanji without actual writing nor intentional mental writing. Activation associated with actual writing was shown in the contralateral primary sensorimotor cortex and the SMA proper. These results suggested that retrieval of kanji would share the neural basis of motor representation with writing of kanji except for regions directly working for motor output. PMID- 10363948 TI - The phenotypic characteristics of heterozygous reeler mouse. AB - Histological and behavioral traits are associated with reelin (Reln) haplo insufficiency in heterozygous reeler mouse (rl+/-). These phenotypic traits are an approximately 50% decrease of brain Reln mRNA and Reln protein, an accumulation of nicotinamide-adenine dinucleotide phosphate-diaphorase (NADPH-d) positive neurons in subcortical white matter, an age-dependent decrease in prepulse inhibition of startle (PPI), and neophobic behavior on the elevated plus maze. Possible analogies between these rl+/- phenotypic traits and signs of psychosis vulnerability are discussed. PMID- 10363950 TI - Metabotropic glutamate receptor mGluR2/3 immunoreactivity in the mouse superior colliculus: co-localization with calbindin D28K. AB - We have studied the distribution of mGluR2/3 in the mouse superior colliculus (SC) with antibody immunocytochemistry and the effect of enucleation on this distribution. We also compared this labeling to that for calbindin D28K. Anti mGluR2/3-immunoreactive (IR) cells formed distinctive laminar patterns within the lower optic and upper intermediate gray layers. By contrast, anti-calbindin D28K IR cells formed obvious laminar patterns in three layers: one within the zonal and upper superficial gray layers, a second within the optic and intermediate gray layers, and the third within the deep gray layer. The distribution of mGluR2/3-IR cells thus matches the second layer of calbindin D28K cells. Two color immunofluorescence revealed that more than half (52.5%) of mGluR2/3-IR cells were also labeled with antibody to calbindin D28K. The majority of mGluR2/3 IR cells were small to medium-sized round/oval or stellate cells. Immunoreactivity for mGluR2/3 was clearly reduced in the contralateral SC following unilateral enucleation. The present results show that mGluR2/3 has a unique cellular sublaminar organization in SC that includes some calbindin D28K IR cells. The effects of enucleation suggest that the retinal projection may control the expression of mGluR2/3 in some cells in the mouse SC. PMID- 10363951 TI - Astrocytes induce several blood-brain barrier properties in non-neural endothelial cells. AB - The passage of immunocompetent cells across the blood-brain barrier (BBB) is regulated at the level of the cerebral capillaries which have specific morphological and biochemical properties. We have developed and characterized an in vitro model of the BBB using immortalized human endothelial cells (ECV 304) induced by rat astrocytes. In this model, endothelial cells are attached together by continuous intercellular junctions with numerous tight junctions, develop a permeability barrier having a significant transcellular electrical resistance, possess high activities of gamma-glutamyl transpeptidase (gamma-GTP) and express the brain-type glucose transporter 1 (GLUT-1). These parameters are also characteristic of brain capillary endothelial cells. Under the culture conditions used, the ECV 304 cells express the intercellular adhesion molecule-1 (ICAM-1) on the external plasma membrane at concentrations which could permit lymphocyte adhesion to be studied. PMID- 10363952 TI - Genetic evidence for oxidative stress in Alzheimer's disease. AB - The cause and proximal consequences of Alzheimer's disease (AD), a progressive debilitating dementia remain largely unknown. Nonetheless an increasing number of genetic risk factors, including most recently bleomycin hydrolase, have been shown to be associated with the disease, offering the hope of revealing the mechanism of disease pathogenesis. Here we show that bleomycin hydrolase, known to be induced in an oxidative environment, is specifically increased in neurons marked for degeneration in AD. These findings support a key proximal role for bleomycin hydrolase, and oxidative stress in AD. PMID- 10363954 TI - Kinetics of polymorphonuclear neutrophil infiltration after a traumatic brain injury in rat. AB - The aim of our study was to assess polymorphonuclear neutrophil infiltration into the injured parenchyma after a traumatic brain injury (TBI). Myeloperoxidase (MPO) activity was assayed on the hippocampus, temporal and parietal cortex 6, 24, 48, 72, and 120 h post-trauma. MPO activity occurred in these structures from 6 h post-trauma and was maximum at 24-48 h. It was resolved by 72 h in the hippocampus and the parietal cortex, but persisted in the temporal cortex until 120 h after trauma. This suggests that neutrophil infiltration is a delayed phenomenon in the physiopathology of TBI. Considering that a large therapeutic window may be crucial in the management of TBI, inhibition of neutrophil infiltration needs to be further investigated following cerebral trauma. PMID- 10363953 TI - Unilateral sciatic nerve injury stimulates contralateral nerve regeneration. AB - Axonal outgrowth in tissue cultures was measured to determine whether unilateral peripheral nerve injuries affect contralateral nerve regeneration. The right sciatic nerves of young male Wistar rats were cut at mid-thigh level. Sham operation as a control was limited to the exposure of the nerve without cutting. At day 6 post-surgery, bilateral L5 dorsal root ganglia (DRG) with attached nerve stumps were resected and cultured. Axonal outgrowth from the nerve stump was measured in situ. The contralateral preparations showed longer outgrowths than controls. Therefore the conditioning effect was not merely restricted to the ipsilateral neurons but also affected undamaged sensory neurons of the contralaretal DRG. PMID- 10363955 TI - In mice tonic estrogen replacement therapy improves non-spatial and spatial memory in a water maze task. AB - We investigated the effects of estrogen replacement therapy on water maze non spatial and spatial navigation in mice. Three groups of mice were ovariectomized and two of these groups being implanted with s.c. pellets that produce blood levels of estrogen close to those found in estrous (estrogen low, 75-100 pg/ml blood) or proestrous (estrogen high, 300-400 pg/ml). The behavioral assessment was initiated 7 days after pellet implantation. Non-spatial navigation to a clearly visible platform was stimulated by low and high levels of estrogen. However, spatial navigation to a hidden platform was improved by low estrogen levels. We found that estrogen improves two different types of memory processes that depend on striatal (non-spatial navigation) and hippocampal (spatial) memory systems. PMID- 10363956 TI - Categorical and coordinate spatial relations: fMRI evidence for hemispheric specialization. AB - Functional magnetic resonance imaging (fMRI) was applied to determine the involvement of the angular gyri in the processing of categorical and coordinate spatial relations. In a categorical task, subjects were asked to judge whether a dot was presented above or below a horizontal line. In a coordinate task, they were asked to judge whether or not the distance between the dot and the bar was within a reference distance. Results showed stronger activation of the left than of the right angular gyrus in the categorical task, and stronger activation, initially, of the right than of the left angular gyrus in the coordinate task. In addition, in the latter task, the involvement of the right angular gyrus decreased with practice while that of the left angular gyrus increased. These results are interpreted in terms of the development of new categorical representations with practice in the coordinate task. PMID- 10363957 TI - Short-term memory for colour following posterior hemispheric lesions in man. AB - Short-term memory for colour was studied in five patients with circumscribed posterior hemispheric lesions. It was impaired independently of colour discrimination in one and more than colour discrimination in two patients. Two patients were normal in colour short-term memory, one with normal and one with deficient colour discrimination performance. Deficient performance in colour short-term memory was associated with bilateral lesions of the inferior occipitotemporal junction including the lateral part of the fusiform gyrus or with a unilateral lesion of the left parieto-occipital convexity. An additional colour constancy deficit was found in the former but not the latter condition. Thus, colour short-term memory can be affected independently of colour discrimination or colour constancy, and may depend on at least two distinct neural circuits. PMID- 10363958 TI - Impact of arthritis and other rheumatic conditions on the health-care system- United States, 1997. AB - Arthritis and other rheumatic conditions are the leading cause of disability in the United States, affecting approximately 43 million persons and costing $65 billion in 1992. By 2020, these numbers will increase as the population ages. This report examines several measures of the impact of arthritis on the U.S. health-care system; the findings indicate that arthritis and other rheumatic conditions have a large impact on hospitalizations, ambulatory-care visits, and home health care, with women accounting for most of this impact and all persons aged <65 years accounting for a substantial portion. PMID- 10363959 TI - Mental retardation following diagnosis of a metabolic disorder in children aged 3 10 years--metropolitan Atlanta, Georgia, 1991-1994. AB - One of the largest population-based disease intervention programs in the United States is newborn metabolic screening. Since the mid- to late 1970s, newborns have been screened routinely for one or more metabolic disorders. The goal of early identification and treatment of metabolic disorders is prevention of the serious medical and developmental consequences of the disorders (e.g., mental retardation [MR]). Despite this goal, the United States has no mechanism for systematic surveillance of the developmental status of children who screen positive for and subsequently have metabolic disorders diagnosed. To determine the number of selected developmental disabilities attributable to metabolic disorders detected by newborn screening, CDC conducted a preliminary investigation of children with developmental disabilities and metabolic disorders in the metropolitan Atlanta area using data from the Metropolitan Atlanta Developmental Disabilities Surveillance Program (MADDSP). This report summarizes the results of this investigation, which indicate that newborn screening is highly effective in reducing the burden of MR associated with these disorders. PMID- 10363961 TI - Assessment of public health computer readiness for 2000--United States, 1999. AB - Computer software, equipment, and other devices that contain embedded microchips that store and process dates may use two-digit years (e.g., 99 for 1999) to reduce data entry burden and save electronic storage space; these devices may not work properly when the year 2000 (Y2K) arrives. Many aspects of health-care delivery, public health surveillance and research, and critical infrastructure components (e.g., utilities and transportation services) depend on vulnerable computers. To ensure that critical public health functions will not be compromised because of Y2K problems, CDC assessed state public health agency readiness for Y2K. This report describes the findings of the assessment, which indicate that state health agencies that responded are substantially ready for Y2K and plan to reach full readiness in 1999. PMID- 10363960 TI - Patients' reports of counseling on mammography screening by health-care providers -North Carolina, 1997. AB - Regular mammography screening combined with timely and appropriate treatment can reduce mortality from breast cancer by 30% in women aged 50-69 years and 16% in women aged 40-49 years. A physician's recommendation has been strongly associated with a patient having a mammogram. This report analyzes data collected during 1997 in North Carolina as part of the Behavioral Risk Factor Surveillance System (BRFSS), which indicated that 23% of women aged > or =40 years who had had a routine physical examination during the 2 years preceding the survey did not recall having a discussion about mammography with a health-care provider. PMID- 10363962 TI - 22nd annual Bristol-Myers Squibb Award for Distinguished Achievement in Cancer Research. Isaiah J. Fidler. PMID- 10363963 TI - Selection for androgen receptor mutations in prostate cancers treated with androgen antagonist. AB - The role of androgen receptor (AR) mutations in androgen-independent prostate cancer (PCa) was determined by examining AR transcripts and genes from a large series of bone marrow metastases. Mutations were found in 5 of 16 patients who received combined androgen blockade with the AR antagonist flutamide, and these mutant ARs were strongly stimulated by flutamide. In contrast, the single mutant AR found among 17 patients treated with androgen ablation monotherapy was not flutamide stimulated. Patients with flutamide-stimulated AR mutations responded to subsequent treatment with bicalutamide, an AR antagonist that blocks the mutant ARs. These findings demonstrate that AR mutations occur in response to strong selective pressure from flutamide treatment. PMID- 10363964 TI - Role of the alternative INK4A proteins in human keratinocyte senescence: evidence for the specific inactivation of p16INK4A upon immortalization. AB - The INK4A locus on human chromosome 9p21 encodes two genes that have been implicated in replicative senescence and tumor suppression, p16INK4A and p14ARF. In contrast to p16INK4A, which is up-regulated to high levels, we were unable to detect p14ARF protein in senescent human keratinocytes. Also, p53, an established target of p14ARF, did not increase, suggesting that p14ARF is not instrumental in human keratinocyte senescence. In neoplastic keratinocyte cultures, p16INK4A inactivation was invariably associated with the immortal phenotype, and there was evidence for the inactivation of p16INK4A, independent of p14ARF, in 6 of 10 lines that lacked large homozygous deletions. In contrast, we failed to detect exon 1beta mutations or p16INK4A-independent deletions. These results emphasize the previously proposed role for p16INK4A in human keratinocyte senescence but do not rule out a supporting role for p14ARF inactivation. PMID- 10363965 TI - Endogenous apurinic/apyrimidinic sites in genomic DNA of mammalian tissues. AB - Apurinic/apyrimidinic (AP) sites are one of the most frequent lesions in DNA. Using a highly sensitive slot blot assay, we determined the number and condition of endogenous AP sites in normal tissues of rats and human liver. The number of AP sites (50,000-200,000 per mammalian cell) was greatest in brain, followed by colon and heart, and then liver, lung, and kidney. The majority of endogenous AP sites were cleaved 5' to the AP site. These data suggest that removal of the deoxyribosyl phosphate moiety is the rate-limiting step in base excision and AP site repair in vivo. PMID- 10363966 TI - A new cis element is involved in the HER2 gene overexpression in human breast cancer cells. AB - The HER2 proto-oncogene product is overexpressed in 30% of breast cancers, and this correlates with poor prognosis. Increased levels of HER2 mRNA in breast cancer cell lines result from increased gene transcription. We report the identification of a new 17-bp-long cis sequence located between positions -506 and -489 from the transcription start site. This sequence is recognized by a trans-activating factor that we tentatively named HER2 transcription factor (HTF). This factor, involved in the increased transcription of the HER2 gene in the BT-474 mammary tumor cells, has a molecular weight of about Mr 50,000. HTF can also bind, but with a lower affinity, to a related cis sequence present in the epidermal growth factor receptor promoter. Interestingly, the HTF binding activity is high in nuclear extracts from several mammary tumor cells overexpressing the HER2 gene. PMID- 10363967 TI - Antifolate resistance mediated by the multidrug resistance proteins MRP1 and MRP2. AB - Transfection of multidrug resistance proteins (MRPs) MRP1 and MRP2 in human ovarian carcinoma 2008 cells conferred a marked level of resistance to short-term (1-4 h) exposure to the polyglutamatable antifolates methotrexate (MTX; 21-74 fold), ZD1694 (4-138-fold), and GW1843 (101-156-fold). Evidence for MRP-mediated antifolate efflux relies upon the following findings: (a) a 2-3.3-fold lower accumulation of [3H]MTX and subsequent reduced formation of long-chain polyglutamate forms of MTX; (b) reversal of MTX resistance by probenecid in both transfectants, and (c) ATP-dependent uptake of [3H]MTX in inside-out vesicles of MRP1 and MRP2 transfectants. This report provides a mechanistic basis for resistance to polyglutamatable antifolates through an MRP-mediated drug extrusion. PMID- 10363968 TI - Recombinant virus vaccination against "self" antigens using anchor-fixed immunogens. AB - To study the induction of anti-"self" CD8+ T-cell reactivity against the tumor antigen gp100, we used a mouse transgenic for a chimeric HLA-A*0201/H-2 Kb molecule (A2/Kb). We immunized the mice with a recombinant vaccinia virus encoding a form of gp100 that had been modified at position 210 (from a threonine to a methionine) to increase epitope binding to the restricting class I molecule. Immunogens containing the "anchor-fixed" modification elicited anti-self CD8+ T cells specific for the wild-type gp100(209-217) peptide pulsed onto target cells. More important, these cells specifically recognized the naturally presented epitope on the surface of an A2/Kb-expressing murine melanoma, B16. These data indicate that anchor-fixing epitopes could enhance the function of recombinant virus-based immunogens. PMID- 10363969 TI - Gain of Bcl-2 is more potent than bax loss in regulating mammary epithelial cell survival in vivo. AB - The impact of gain of Bcl-2 function on mammary epithelial cell survival was compared with loss of Bax function during the two stages of mammary gland involution. Bcl-2 gain of function reduced apoptosis 50% during the first stage and increased cell survival 70% during the second stage. Complete loss of Bax reduced apoptosis by 20% during the first stage without second stage effect. Partial loss of Bax was ineffective but increased cell survival 2.4-fold when combined with Bcl-2 gain. Gain of Bcl-2 function is more potent than loss of Bax function in regulating mammary epithelial cell survival in vivo. PMID- 10363970 TI - Increased level of exon 12 alternatively spliced BRCA2 transcripts in tumor breast tissue compared with normal tissue. AB - The breast cancer susceptibility gene BRCA2 is expressed in a wide range of tissues as an 11-kb mRNA transcript encoding a 3418-amino acid protein, which is involved in the response to DNA damage. To obtain a better molecular characterization of BRCA2 expression in breast tissue, we analyzed full-length BRCA2 mRNA by means of reverse transcriptase-PCR with a panel of primer pairs encompassing the entire cDNA sequence. We report the identification of an exon 12 alternatively spliced BRCA2 transcript (delta12-BRCA2) in normal human breast tissue, in a wide variety of other normal human tissues, and in several mouse tissues. The deletion observed in this transcript (96 bp) preserves the open reading frame, and translation of the transcript would result in a BRCA2 isoform lacking 32 amino acids between codons 2280 and 2311. The analysis of matched normal and primary tumor breast tissues from 12 patients showed that the expression level of the delta12-BRCA2 transcript was higher in 4 of 12 (33%) tumor tissues compared with their normal breast tissues. Overproduction of the delta12-BRCA2 variant was associated with steroid receptor-negative tumors (P = 0.0005). These data suggest that the mechanisms generating the BRCA2 mRNA variant exist in normal breast tissue and may be dysregulated in steroid receptor negative breast tumor tissues. PMID- 10363971 TI - Regulation of Akt/PKB activity, cellular growth, and apoptosis in prostate carcinoma cells by MMAC/PTEN. AB - Understanding the functional roles of the molecular alterations that are involved in the oncogenesis of prostate cancer, the second most frequent cause of cancer related deaths among men in the United States is the focus of numerous investigations. To examine the possible significance of alterations associated with the tumor suppressor gene, MMAC/PTEN, in prostate carcinoma, the biological and biochemical effects of MMAC/PTEN expression were examined in LNCaP cells, which are devoid of a functional gene product. Acute expression of MMAC/PTEN via an adenoviral construct resulted in a dose-dependent and specific inhibition of Akt/PKB activation, consistent with the phosphatidylinositol phosphatase activity of MMAC/PTEN. MMAC/PTEN expression induced apoptosis in LNCaP cells, although to a lesser extent than that observed with p53 via an adenoviral construct. However, MMAC/PTEN expression produced a growth inhibition that was significantly greater than that achieved with p53. Overexpression of Bcl-2 in LNCaP cells blocked MMAC/PTEN- and p53-induced apoptosis but not the growth-suppressive effects of MMAC/ PTEN, suggesting that the growth regulatory effects of MMAC/PTEN involve multiple pathways. These studies further implicate the loss of MMAC/PTEN as a significant event in prostate cancer and suggest that reintroduction of MMAC/PTEN into deficient prostate cancer cells may have therapeutic implications. PMID- 10363972 TI - XRCC1 polymorphisms: effects on aflatoxin B1-DNA adducts and glycophorin A variant frequency. AB - Hereditary genetic defects in DNA repair lead to increased risk of cancer. Polymorphisms in several DNA repair genes have been identified; however, the impact on repair phenotype has not been elucidated. We explored the relationship between polymorphisms in the DNA repair enzyme, XRCC1 (codons 194, 280, and 399), and genotoxic end points measured in two populations: (a) placental aflatoxin B1 DNA (AFB1-DNA) adducts in a group of Taiwanese maternity subjects (n = 120); and (b) somatic glycophorin A (GPA) variants in erythrocytes from a group of North Carolina smokers and nonsmokers (n = 59). AFB1-DNA adducts were measured by ELISA, and erythrocyte GPA variant frequency (NN and NO) was assessed in MN heterozygotes with a flow cytometric assay. XRCC1 genotypes were identified by PCR-RFLPs. The XRCC1 399Gln allele was significantly associated with higher levels of both AFB1-DNA adducts and GPA NN mutations. Individuals with the 399Gln allele were at risk for detectable adducts (odds ratio, 2.4; 95% confidence interval, 1.1-5.4; P = 0.03). GPA NN variant frequency was significantly higher in 399Gln homozygotes (19.6 x 10(-6)) than in Gln/Arg heterozygotes (11.4 x 10( 6); P < 0.05) or Arg/Arg homozygotes (10.1 x 10(-6); P = 0.01). No significant effects were observed for other XRCC1 polymorphisms. These results suggest that the Arg399Gln amino acid change may alter the phenotype of the XRCC1 protein, resulting in deficient DNA repair. PMID- 10363973 TI - p53 directly enhances rejoining of DNA double-strand breaks with cohesive ends in gamma-irradiated mouse fibroblasts. AB - The p53 gene regulates the cell cycle response to DNA damage, which may allow time for adequate DNA repair. We asked whether p53 could directly increase the repair of defined double-strand breaks (DSBs) by nonhomologous end-joining in gamma-irradiated mouse embryonic fibroblasts with differing p53 status. By using an episomal plasmid reactivation assay, we found that presence of wild-type p53 enhanced rejoining of DSBs with short complementary ends of single-stranded DNA. p53 appeared to be directly involved in this regulation, because rejoining enhancement was dependent on the presence of nonspecific DNA binding activity as mediated by the COOH-terminal domain and was independent of transactivating function. We hypothesize that tumor cells lacking p53 and normal cells with wild type p53 may use different pathways for repair of radiation-induced DSBs. PMID- 10363974 TI - Discovery and initial characterization of the paullones, a novel class of small molecule inhibitors of cyclin-dependent kinases. AB - Analysis of the National Cancer Institute Human Tumor Cell Line Anti-Cancer Drug Screen data using the COMPARE algorithm to detect similarities in the pattern of compound action to flavopiridol, a known inhibitor of cyclin-dependent kinases (CDKs), has suggested several possible novel CDK inhibitors. 9-Bromo-7,12-dihydro indolo[3,2-d][1]benzazepin-6(5H)-one, NSC-664704 (kenpaullone), is reported here to be a potent inhibitor of CDK1/cyclin B (IC50, 0.4 microM). This compound also inhibited CDK2/cyclin A (IC50, 0.68 microM), CDK2/cyclin E (IC50, 7.5 microM), and CDK5/p25 (IC50, 0.85 microM) but had much less effect on other kinases; only c-src (IC50, 15 microM), casein kinase 2 (IC50, 20 microM), erk 1 (IC50, 20 microM), and erk 2 (IC50, 9 microM) were inhibited with IC50s less than 35 microM. Kenpaullone acts by competitive inhibition of ATP binding. Molecular modeling indicates that kenpaullone can bind in the ATP binding site of CDK2 with residue contacts similar to those observed in the crystal structures of other CDK2-bound inhibitors. Analogues of kenpaullone, in particular 10-bromopaullone (NSC-672234), also inhibited various protein kinases including CDKs. Cells exposed to kenpaullone and 10-bromopaullone display delayed cell cycle progression. Kenpaullone represents a novel chemotype for compounds that preferentially inhibit CDKs. PMID- 10363975 TI - Identification and characterization of human MT5-MMP, a new membrane-bound activator of progelatinase a overexpressed in brain tumors. AB - A cDNA encoding a new member of the membrane-type (MT) matrix metalloproteinase (MMP) family has been identified and cloned from a human brain cDNA library. The isolated cDNA encodes a polypeptide of 645 amino acids that displays a similar domain organization as other MMPs, including a predomain with the activation locus, a zinc-binding site, and a hemopexin domain. The deduced amino acid sequence contains a COOH-terminal extension, rich in hydrophobic residues and similar in size to the equivalent domains identified in MT-MMPs. Immunofluorescence and Western blot analysis of COS-7 cells transfected with the isolated cDNA revealed that the encoded protein is localized in the plasma membrane. On the basis of these features, this novel human MMP has been called MT5-MMP because it represents the fifth member of the MT-MMP subfamily of MMPs. Fluorescent in situ hybridization experiments showed that the human MT5-MMP gene (MMP-24) maps to 20q11.2, a region frequently amplified in tumors from diverse sources. Northern blot analysis demonstrated that MT5-MMP is predominantly expressed in brain, kidney, pancreas, and lung. In addition, MT5-MMP transcripts were detected at high levels compared to normal brain tissue in a series of brain tumors, including astrocytomas and glioblastomas. The catalytic domain of MT5 MMP, produced in Escherichia coli as a fusion protein with glutathione S transferase, exhibits a potent proteolytic activity against progelatinase A, leading to the generation of the Mr 62,000 active form of this enzyme. These data suggest that MT5-MMP may contribute to the activation of progelatinase A in tumor tissues, in which it is overexpressed, thereby facilitating tumor progression. PMID- 10363976 TI - Transduced p16INK4a peptides inhibit hypophosphorylation of the retinoblastoma protein and cell cycle progression prior to activation of Cdk2 complexes in late G1. AB - Progression of cells through the G1 phase of the cell cycle requires cyclin D:Cdk4/6 and cyclin E:Cdk2 complexes; however, the duration and ordering of these complexes remain unclear. To address this, we synthesized a peptidyl mimetic of the Cdk4/6 inhibitor, p16INK4a that contained an NH2-terminal TAT protein transduction domain. Transduction of TAT-p16 wild-type peptides into cells resulted in the loss of active, hypophosphorylated pRb and elicited an early G1 cell cycle arrest, provided cyclin E:Cdk2 complexes were inactive. We conclude that cyclin D:Cdk4/6 activity is required for early G1 phase cell cycle progression up to, but not beyond, activation of cyclin E:Cdk2 complexes at the restriction point and is thus nonredundant with cyclin E:Cdk2 in late G1. PMID- 10363977 TI - Competitive and noncompetitive inhibition of the DNA-dependent protein kinase. AB - The DNA-dependent protein kinase (DNA-PK) is a serine/threonine protein kinase that is involved in mammalian DNA double-strand break repair. The catalytic subunit of DNA-PK (DNA-PKcs) shares sequence homology in its kinase domain with phosphatidylinositol (PI) 3-kinase. Here, we provide a detailed kinetic analysis of DNA-PK inhibition by the PI 3-kinase inhibitor wortmannin and demonstrate this inhibition to be of a noncompetitive nature, with a Ki of 120 nM. Another inhibitor of PI 3-kinase. LY294002, its parent compound, quercetin, and other derivatives have also been studied. These chemicals are competitive inhibitors of DNA-PK, with LY294002 having a Ki of 6.0 microM. Using an antibody to wortmannin, we found that this compound binds covalently to the kinase domain of DNA-PKcs both in vitro and in vivo. Binding of wortmannin to the active site of DNA-PKcs is inhibited by ATP but not by a peptide substrate. Furthermore, wortmannin is able to bind to DNA-PKcs independently of Ku, and it is not stimulated by the presence of DNA. This suggests that the ATP binding site of DNA-PKcs is open constitutively and that DNA activation of the kinase is mediated via another mechanism. PMID- 10363978 TI - In vivo mutagenicity and hepatocarcinogenicity of 2-amino-3,8-dimethylimidazo[4,5 f]quinoxaline (MeIQx) in bitransgenic c-myc/lambda lacZ mice. AB - 2-Amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) is a mutagenic and carcinogenic heterocyclic amine found in cooked meat. Hepatic DNA adduct formation, in vivo mutagenicity, and hepatocarcinogenicity of MeIQx were examined in mice harboring the lacZ mutation reporter gene (Muta mice) and bitransgenic mice overexpressing the c-myc oncogene. C57Bl/lambda lacZ and bitransgenic c-myc (albumin promoter)/lambda lacZ mice were bred and weaned onto an American Institute of Nutrition-76-based diet containing 0.06% (w/w) MeIQx or onto control diet. After 30 weeks on diet, only male bitransgenic mice on MeIQx developed hepatocellular carcinoma (100% incidence). By 40 weeks, hepatic tumor incidence was 100%/75% (17%/0%) and 44%/17% (0%/0%) in male c-myc/lambda lacZ and C57Bl/lambda lacZ mice who were given MeIQx (or control) diet, respectively, supporting a synergism between MeIQx and c-myc overexpression in hepatocarcinogenesis. At either time point, mutant frequency in the lacZ gene was at least 40-fold higher in MeIQx-treated mice than in control mice of either strain. These findings suggest that MeIQx-induced hepatocarcinogenesis is associated with MeIQx-induced mutations. Elevated mutant frequency in MeIQx treated mice also occurred concomitant with the formation of MeIQx-guanine adducts, as detected by the 32P-postlabeling assay. Irrespective of strain or diet, sequence analysis of the lacZ mutants from male mouse liver showed that the principal sequence alterations were base substitutions at guanine bases. Adenine mutations, however, were detected only in animals on control diet. MeIQx-fed mice harboring the c-myc oncogene showed a 1.4-2.6-fold higher mutant frequency in the lacZ gene than mice not carrying the transgene. Although there was a trend toward higher adduct levels in c-myc mice, MeIQx-DNA adduct levels were not significantly different between c-myc/lambda lacZ and C57Bl/lambda lacZ mice after 30 weeks on diet. Thus, it seemed that factors in addition to MeIQx-DNA adduct levels, such as the enhanced rate of proliferation associated with c-myc overexpression, may have accounted for a higher mutant frequency in c-myc mice. In the control diet groups, the lacZ mutant frequency was significantly higher in c-myc/lambda lacZ mice than in C57Bl/lambda lacZ mice. The findings are consistent with the notion that c-myc overexpression is associated with an increase in mutagenesis. The mechanism for the synergistic effects of c-myc overexpression on MeIQx hepatocarcinogenicity seems to involve an enhanced expression of MeIQx-induced mutations. PMID- 10363979 TI - Bone marrow pre-B-1 (Bomb1): a quantitative trait locus inducing bone marrow pre B-cell expansion in lymphoma-prone SL/Kh mice. AB - Abnormalities of regulatory genes in early B-cell development often lead to lymphomagenesis. Our previous study showed that there is an abnormal transient expansion of bone marrow (BM) pre-B cells in lymphoma-prone SL/Kh strain mice. Such expansion is a genetic property of SL/Kh stem cells rather than BM microenvironments. Using the percentage of BP1+ B220+ pre-B cells in total BM lymphoid cells as a quantitative parameter, we studied the genetic control of BM pre-B cells in 159 F2 offspring of crosses between SL/Kh and NFS/N mice and 334 back-crosses to SL/Kh mice. A highly significant quantitative trait locus was identified on the distal segment of chromosome 3, showing logarithm of odds scores of 22.7 in the F2 cohort and 10.7 in back-cross mice. This quantitative trait locus, named bone marrow pre-B-1, colocalized with lymphoid enhancer factor 1, which encodes a high mobility group DNA-binding protein that is expressed in T and pre-B cells. PMID- 10363980 TI - Resveratrol increases nitric oxide synthase, induces accumulation of p53 and p21(WAF1/CIP1), and suppresses cultured bovine pulmonary artery endothelial cell proliferation by perturbing progression through S and G2. AB - Epidemiological studies have shown that the regular consumption of red wine may in part account for the apparent compatibility of a high fat diet with a low incidence of coronary atherosclerosis. This phenomenon, commonly referred to as the French paradox, may be associated with red wine constituents that exhibit tumor-preventive properties as well as inhibit reactions that increase the risk of coronary heart disease. Here we show that resveratrol, a polyphenol in red wine, induces nitric oxide synthase, the enzyme responsible for the biosynthesis of NO, in cultured pulmonary artery endothelial cells, suggesting that resveratrol could afford cardioprotection by affecting the expression of nitric oxide synthase. We also show that resveratrol inhibits the proliferation of pulmonary artery endothelial cells, which, based on flow cytometric analysis, correlates with the suppression of cell progression through S and G2 phases of the cell cycle. Western blot analysis and immunocytochemical protein detection combined with multiparameter flow cytometry further demonstrate that the perturbed progression through S and G2 phases is accompanied by an increase in the expression of tumor suppressor gene protein p53 and elevation of the level of cyclin-dependent kinase inhibitor p21(WAF1/CIP1). All of the observed effects of resveratrol, including induction of apoptosis at its higher concentration, are also compatible with its putative chemopreventive and/or antitumor activity. PMID- 10363981 TI - Defective control of apoptosis and mitotic spindle checkpoint in heterozygous carriers of ATM mutations. AB - Ataxia telangiectasia (AT) carrier-derived lymphoblastoid cell lines (AT LCLs/hetero) with suboptimal ATM protein expression were examined for the regulation of radiosensitivity, apoptosis, and mitotic spindle checkpoint in response to DNA-damaging agents. Although AT-LCLs/hetero showed intermediate radiation sensitivity, as determined by clonogenic assay, they were resistant to early-onset apoptosis, as much as AT patient-derived LCLs (AT-LCLs/homo). Furthermore, two of three AT-LCLs/hetero showed defective mitotic spindle checkpoint control in response to X-ray irradiation, which is a recently characterized biological feature in AT-LCLs/homo. Our findings indicate that carriers of ATM mutation have biological abnormalities due to haploinsufficiency of ATM protein or dominant-negative effect of mutant ATM protein. Thus, although it is still controversial whether ATM mutation carriers are at higher risk for cancer during adulthood, our findings based on in vitro biological indicators support the notion that at least some of such carriers are at a higher risk for cancer development than those without ATM mutation. Our findings may help to reevaluate epidemiological studies on cancer susceptibility in AT carriers. PMID- 10363983 TI - Proteasome inhibitors: a novel class of potent and effective antitumor agents. AB - The ubiquitin-proteasome pathway plays a critical role in the regulated degradation of proteins involved in cell cycle control and tumor growth. Dysregulating the degradation of such proteins should have profound effects on tumor growth and cause cells to undergo apoptosis. To test this hypothesis, we developed a novel series of proteasome inhibitors, exemplified by PS-341, which we describe here. As determined by the National Cancer Institute in vitro screen, PS-341 has substantial cytotoxicity against a broad range of human tumor cells, including prostate cancer cell lines. The PC-3 prostate cell line was, therefore, chosen to further examine the antitumor activity of PS-341. In vitro, PS-341 elicits proteasome inhibition, leading to an increase in the intracellular levels of specific proteins, including the cyclin-dependent kinase inhibitor, p21. Moreover, exposure of such cells to PS-341 caused them to accumulate in the G2-M phase of the cell cycle and subsequently undergo apoptosis, as indicated by nuclear condensation and poly(ADP-ribose) polymerase cleavage. Following weekly i.v. treatment of PS-341 to mice bearing the PC-3 tumor, a significant decrease (60%) in tumor burden was observed in vivo. Direct injection of PS-341 into the tumor also caused a substantial (70%) decrease in tumor volume with 40% of the drug-treated mice having no detectable tumors at the end of the study. Studies also revealed that i.v. administration of PS-341 resulted in a rapid and widespread distribution of PS-341, with highest levels identified in the liver and gastrointestinal tract and lowest levels in the skin and muscle. Modest levels were found in the prostate, whereas there was no apparent penetration of the central nervous system. An assay to follow the biological activity of the PS 341 was established and used to determine temporal drug activity as well as its ability to penetrate tissues. As such, PS-341 was shown to penetrate PC-3 tumors and inhibit intracellular proteasome activity 1.0 h after i.v. dosing. These data illustrate that PS-341 not only reaches its biological target but has a direct effect on its biochemical target, the proteasome. Importantly, the data show that inhibition of this target site by PS-341 results in reduced tumor growth in murine tumor models. Together, the results highlight that the proteasome is a novel biochemical target and that inhibitors such as PS-341 represent a unique class of antitumor agents. PS-341 is currently under clinical evaluation for advanced cancers. PMID- 10363982 TI - Fibroblast growth factor 2 retargeted adenovirus has redirected cellular tropism: evidence for reduced toxicity and enhanced antitumor activity in mice. AB - Adenovirus (Ad) have been used as vectors to deliver genes to a wide variety of tissues. Despite achieving high expression levels in vivo, Ad vectors display normal tissue toxicity, transient expression, and antivector immune responses that limit therapeutic potential. To circumvent these problems, several retargeting strategies to abrogate native tropism and redirect Ad uptake through defined receptors have been attempted. Despite success in cell culture, in vivo results have generally not shown sufficient selectivity for target tissues. We have previously identified (C. K. Goldman et al., Cancer Res., 57: 1447-1451, 1997) the fibroblast growth factor (FGF) ligand and receptor families as conferring sufficient specificity and binding affinity to be useful for targeting DNA in vivo. In the present studies, we retargeted Ad using basic FGF (FGF2) as a targeting ligand. Cellular uptake is redirected through high-affinity FGF receptors (FGFRs) and not the more ubiquitous lower-affinity Ad receptors. Initial in vitro experiments demonstrated a 10- to 100-fold increase in gene expression in numerous FGFR positive (FGFR+) cell lines using FGF2-Ad when compared with Ad. To determine whether increased selectivity could be detected in vivo, FGF2-Ad was administered i.v. to normal mice. FGF2-Ad demonstrates markedly decreased hepatic toxicity and liver transgene expression compared with Ad treatment. Importantly, FGF2-Ad encoding the herpes simplex virus thymidine kinase (TK) gene transduces Ad-resistant FGFR+ tumor cells both ex vivo and in vivo, which results in substantially enhanced survival (180-260%) when the prodrug ganciclovir is administered. Because FGFRs are up-regulated on many types of malignant or injured cells, this broadly useful method to redirect native Ad tropism and to increase the potency of gene expression may offer significant therapeutic advantages. PMID- 10363984 TI - Intravenous administration of ONYX-015, a selectively replicating adenovirus, induces antitumoral efficacy. AB - Replication-incompetent viral vectors are being developed for the gene therapy of cancer. Although some of these may eventually be proven to have significant localized antitumoral activity, none to date have been shown to infect and cause regression of established tumors following i.v. administration. Because cancer is a systemic disease in almost all fatal cases, the lack of i.v. efficacy is a major limitation to treatment with replication-incompetent viral vectors. ONYX 015 (d11520) is an attenuated adenovirus that replicates in and causes selective lysis of cancer cells. We carried out i.v. efficacy and distribution studies in nude mice with s.c. and intraparenchymal tumor xenografts. ONYX-015 infected and replicated efficiently within tumors following i.v. administration. Viral titers in livers were relatively high 3 h after administration but decreased rapidly, becoming undetectable after 24 h. Effective antitumor doses were not associated with hepatic toxicity. Viral replication within tumors was associated with regressions in several tumor models. Selectively replicating viruses like ONYX 015 hold promise as agents to treat metastatic cancer. PMID- 10363985 TI - Evaluation of cell-killing effects of 1-beta-D-arabinofuranosylcytosine and daunorubicin by a new computer-controlled in vitro pharmacokinetic simulation system. AB - An in vitro pharmacokinetic simulation system that can simulate plasma pharmacokinetics was established to evaluate the cytotoxicity of two representative antileukemic agents, 1-beta-D-arabinofuranosylcytosine (ara-C) and daunorubicin. With this system, the survival rate of the cell line K562 treated with ara-C, relative to that of untreated control cells, was 7.1%, as determined by a clonogenic culture technique using a clinically intermediate dose of ara-C (1.0 g/m2; 2-h infusion). When the area under the serum concentration-time curve (AUC) was kept constant at infusion times of 2, 4, 8, and 16 h with a regular dose of ara-C (100-mg/m2 infusion), the relative cell survival rates were 75, 72, 34, and 14%, respectively. In contrast, with the use of a conventional culture system and a constant concentration-time product (C x T), no time-dependent inhibition effect by ara-C was observed, probably due to the ara-C inactivation in the cell-containing culture medium. For example, 93% of the ara-C in the cell suspended medium in the conventional culture system was converted to its inactive form, 1-beta-D-arabinofuranosyluracil, within 16 h after addition of ara-C to the medium. For the simulations of the administration of 50 mg/m2 daunorubicin for 0.5-, 2-, 4-, and 8-h infusions, the relative survival rates [maximal concentrations (Cmaxs)] were 37.4% (0.24 microM), 49.7% (0.089 microM), 72.1% (0.055 microM), and 82.2% (0.032 microM), respectively. With the conventional culture system, the relative survival rates (Cmaxs) following daunorubicin treatment were 7.6% (0.48 microM), 18.6% (0.12 microM), 63.7% (0.06 microM), and 92.0% (0.03 microM) when the drug exposure times were 0.5, 2, 4, and 8 h, respectively, with a constant C x T. When the drug concentration for 90% cell killing by the conventional culture system was plotted against the exposure time on a logarithmic scale, the regression line for daunorubicin had a slope of 0.40, whereas the slope of cis-diamminedichloroplatinum (or cisplatin), a typical AUC-dependent drug, was -0.98. These results suggested that daunorubicin was Cmax dependent rather than AUC dependent. In the simulation system, this Cmax dependency was apparently reduced, probably because of the smaller difference of Cmaxs in the simulation system compared with the conventional system with the constant AUC. Thus, this simulation system can predict the effects of ara-C, daunorubicin, and other antineoplastic agents much more exactly than the conventional culture system in clinical use. PMID- 10363986 TI - High-linear energy transfer (LET) alpha versus low-LET beta emitters in radioimmunotherapy of solid tumors: therapeutic efficacy and dose-limiting toxicity of 213Bi- versus 90Y-labeled CO17-1A Fab' fragments in a human colonic cancer model. AB - Recent studies suggest that radioimmunotherapy (RIT) with high-linear energy transfer (LET) radiation may have therapeutic advantages over conventional low LET (e.g., beta-) emissions. Furthermore, fragments may be more effective in controlling tumor growth than complete IgG. However, to the best of our knowledge, no investigators have attempted a direct comparison of the therapeutic efficacy and toxicity of a systemic targeted therapeutic strategy, using high-LET alpha versus low-LET beta emitters in vivo. The aim of this study was, therefore, to assess the toxicity and antitumor efficacy of RIT with the alpha emitter 213Bi/213Po, as compared to the beta emitter 90Y, linked to a monovalent Fab' fragment in a human colonic cancer xenograft model in nude mice. Biodistribution studies of 213Bi- or 88Y-labeled benzyl-diethylene-triamine-pentaacetate conjugated Fab' fragments of the murine monoclonal antibody CO17-1A were performed in nude mice bearing s.c. human colon cancer xenografts. 213Bi was readily obtained from an "in-house" 225Ac/213Bi generator. It decays by beta- and 440-keV gamma emission, with a t(1/2) of 45.6 min, as compared to the ultra-short lived alpha emitter, 213Po (t(1/2) = 4.2 micros). For therapy, the mice were injected either with 213Bi- or 90Y-labeled CO17-1A Fab', whereas control groups were left untreated or were given a radiolabeled irrelevant control antibody. The maximum tolerated dose (MTD) of each agent was determined. The mice were treated with or without inhibition of the renal accretion of antibody fragments by D lysine (T. M. Behr et al., Cancer Res., 55: 3825-3834, 1995), bone marrow transplantation, or combinations thereof. Myelotoxicity and potential second organ toxicities, as well as tumor growth, were monitored at weekly intervals. Additionally, the therapeutic efficacy of both 213Bi- and 90Y-labeled CO17-1A Fab' was compared in a GW-39 model metastatic to the liver of nude mice. In accordance with kidney uptake values of as high as > or = 80% of the injected dose per gram, the kidney was the first dose-limiting organ using both 90Y- and 213Bi-labeled Fab' fragments. Application of D-lysine decreased the renal dose by >3-fold. Accordingly, myelotoxicity became dose limiting with both conjugates. By using lysine protection, the MTD of 90Y-Fab' was 250 microCi and the MTD of 213Bi Fab' was 700 microCi, corresponding to blood doses of 5-8 Gy. Additional bone marrow transplantation allowed for an increase of the MTD of 90Y-Fab' to 400 microCi and for 213Bi-Fab' to 1100 microCi, respectively. At these very dose levels, no biochemical or histological evidence of renal damage was observed (kidney doses of <35 Gy). At equitoxic dosing, 213Bi-labeled Fab' fragments were significantly more effective than the respective 90Y-labeled conjugates. In the metastatic model, all untreated controls died from rapidly progressing hepatic metastases at 6-8 weeks after tumor inoculation, whereas a histologically confirmed cure was observed in 95% of those animals treated with 700 microCi of 213Bi-Fab' 10 days after model induction, which is in contrast to an only 20% cure rate in mice treated with 250 microCi of 90Y-Fab'. These data show that RIT with alpha emitters may be therapeutically more effective than conventional beta emitters. Surprisingly, maximum tolerated blood doses were, at 5-8 Gy, very similar between high-LET alpha and low-LET beta emitters. Due to its short physical half-life, 213Bi appears to be especially suitable for use in conjunction with fast-clearing fragments. PMID- 10363987 TI - 1,25-Dihydroxycholecalciferol (1,25-D3) inhibits the growth of squamous cell carcinoma and down-modulates p21(Waf1/Cip1) in vitro and in vivo. AB - 1,25-Dihydroxycholecalciferol (1,25-D3) has significant antitumor effects in the murine squamous cell carcinoma (SCC) tumor model in vitro and in vivo. We investigated the basis for this antiproliferative activity and found that, in vitro, 1,25-D3 administration is associated with altered expression of cell cycle regulatory proteins, treatment results in retinoblastoma dephosphorylation, decreased expression of p21(Waf1/Cip1) (p21) mRNA and protein, and increased expression of p27Kip1 (p27) mRNA and protein. Dexamethasone, which acts synergistically with 1,25-D3 to inhibit SCC proliferation, enhanced 1,25-D3 induced down-modulation of p21 without affecting the ability of 1,25-D3 to increase p27 expression. 1,25-D3 did not induce cleavage of poly(ADP-ribose) polymerase. These in vitro data suggest that 1,25-D3 exerts antitumor activity in SCC by perturbing cell cycle progression rather than by inducing apoptosis. In vivo, a 1,25-D3 treatment regimen that results in a decrease in SCC tumor volume is associated with a statistically significant decrease in intratumoral p21 expression. p21 expression is not changed in tumors isolated from control animals or animals treated with a nontherapeutic dose of 1,25-D3. Intratumoral p27 levels were not modulated by 1,25-D3 treatment. Thus, both in vitro and in vivo, 1,25-D3 mediated growth inhibition is associated with p21 down-modulation. PMID- 10363988 TI - Induction of therapeutic T-cell immunity by tumor targeting with soluble recombinant B7-immunoglobulin costimulatory molecules. AB - Tumor targeting with immunomodulatory molecules is an attractive strategy to enhance the host's antitumor response. Expression of CD80 (B7-1) and CD86 (B7-2) costimulatory molecules in tumor cells has proven to be an efficient way to enhance their immunogenicity. Here, we studied the effects of tumor targeting with biotinylated recombinant soluble B7-1- and B7-2 immunoglobulin G molecules (bio-B7-IgG) using a pretargeting approach based on the sequential use of a biotinylated antitumor monoclonal antibody and avidin. Mouse RMA T-lymphoma cells bearing either bio-B7-1-IgG or bio-B7-2-IgG on their surface prime in vitro naive CD8+ CTLs, which are highly effective in adoptive immunotherapy, and induce therapeutic immunity when injected in tumor-bearing animals. In vivo targeting of established RMA tumors with bio-B7-IgG either cures tumor-bearing mice or significantly prolongs their survival. The antitumor response induced by targeted bio-B7-IgG depends on both CD4+ and CD8+ T cells. Moreover, tumor targeting with bio-B7-IgG in vivo is critical for both expansion in lymphoid organs and mobilization into the tumor of tumor-specific CD8+ CTLs. When targeting is performed on poorly immunogenic TS/A mammary adenocarcinoma, only bio-B7-1-IgG primes naive CTLs in vitro and cures or significantly prolongs the survival of tumor-bearing mice in vivo, confirming that the two costimulatory molecules are not redundant with this tumor. Altogether, these data suggest that tumor avidination and targeting with soluble bio-B7-IgG may represent a promising strategy to enhance the antitumor response in the host. PMID- 10363989 TI - Low prevalence of selective human leukocyte antigen (HLA)-A and HLA-B epitope losses in early-passage tumor cell lines. AB - The down-regulation of human leukocyte antigen (HLA) class I molecules, especially the selective down-regulation of certain allelic products, is believed to represent a major mechanism of tumor escape from immune surveillance. In the present report, an original approach is described to precisely evaluate and classify HLA class I epitope losses in 30 cancer patients with malignant melanoma and lung, breast, endometrium, ovary, and colon carcinoma tumors. Early-passage tumor cell lines were established in culture from the corresponding metastatic tumor lesions obtained in each patient. Both the cell lines and the tumor lesions were compared, in their HLA-A and -B expression, to the peripheral blood mononuclear cells (PBMCs) obtained from the same patient (autologous PBMCs). On the basis of HLA-genotyping data, the appropriate monoclonal antibodies identifying mono- and poly-morphic HLA-A and HLA-B epitopes were selected from a panel of 34 antibodies for a total of 24 testable alleles. The selected antibodies were used not only in immunohistochemical assays on cryostatic tumor sections and cytospins of PBMCs but also in quantitative, sensitive flow cytometry assays on early-passage tumor cells and PBMC suspensions. With this latter method, a low overall HLA expression was detected in 26 tumor cell explants and a complete, generalized HLA-A, HLA-B, HLA-C loss in the remaining 4 cases. However, no complete, selective loss of any of the 45 tested HLA-A and HLA B allomorphs was observed. Sequences from all of the HLA class I alleles could be detected at the genomic DNA level in tumor cells and tissues. At variance from the literature and the results of immunohistochemical experiments performed in parallel on the corresponding tumor lesions, the relative proportions of the various HLA epitopes were relatively preserved in each early-passage cell line/PBMC pair, and selective increases, rather than decreases, in the expression of polymorphic HLA epitopes had the highest prevalence and greatest magnitude. Our data suggest an alternative tumor stealth strategy in which up- and down regulation are equally important. This alternative model of tumor-host interaction better fits the available models of tumor cell recognition by CTLs and natural killer cells bearing activatory and inhibitory receptors for HLA-A, HLA-B, HLA-C molecules. PMID- 10363990 TI - Identification of a promiscuous T-cell epitope encoded by multiple members of the MAGE family. AB - One of the major limitations of tumor-specific vaccination is the generation of antigen-loss variants that are able to escape the immune response elicited by a monoantigenic peptide epitope. Here, we report the identification of a new HLA B*3701-restricted epitope shared by four different members of the MAGE family. Peripheral blood lymphocytes isolated from a melanoma patient were stimulated in vitro with the autologous HLA-negative melanoma line transfected with autologous HLA B*3701 molecule. This protocol led to the induction of tumor-specific, B*3701 restricted CTLs specific for a peptide epitope encoded by codons 127-136 of the gene MAGE-1. The same epitope is also encoded by the homologous region of three other members of the MAGE family, MAGE-2, -3, and -6. Consistent with the notion that the peptide encoded by MAGE-1 codons 127-136 is, indeed, processed from the proteins encoded by all four MAGE family members, the CTLs were able to specifically recognize Cos-7 cells cotransfected with HLA-B*3701 and any of these MAGE genes. Moreover, the CTLs also recognized a MAGE-6-positive melanoma line transfected with the B*3701 molecule. These findings allow the inclusion of a new set of tumor patients into clinical cancer vaccination trials. Furthermore, they suggest that some promiscuous peptide epitopes shared by different members of the MAGE family might be less prone to escape the immune response by generation of MAGE antigen loss variants. PMID- 10363991 TI - A PR1-human leukocyte antigen-A2 tetramer can be used to isolate low-frequency cytotoxic T lymphocytes from healthy donors that selectively lyse chronic myelogenous leukemia. AB - We previously showed (E. Clave et al., J. Immunother., 22: 1-6, 1999; J. Molldrem et al., Blood, 88: 2450-2457, 1996) that PR1, a human-lymphocyte-antigen (HLA) A2.1-restricted peptide from proteinase 3, could be used to elicit CTLs from normal individuals. These CTLs showed HLA-restricted cytotoxicity and colony inhibition of myeloid leukemia cells that overexpress proteinase 3. In this study, we constructed a phycoerythrin-labeled PR1-HLA-A2 tetramer to identify PR1 specific CTLs by flow cytometry. No peripheral blood lymphocytes from three HLA 2.1+ donors stained with the tetramer, but, after 20 days in culture with weekly PR1 stimulation, 2-8% became tetramer+. Tetramer staining identified up to 40 fold more PR1-specific CTLs than were identified by limiting dilution analysis and correlated better with lysis of PR1-coated T2 cells (R2 = 0.95 versus R2 = 0.76). Tetramer+ CTLs were memory phenotype (91% CD45RO+), and most (58% CD95+) were activated. Tetramer-sorted allogeneic CTLs produced 83% lysis of HLA-A2.1+ chronic myelogenous leukemia (CML) blasts at an E:T ratio of 2.5:1, compared with 23% lysis by nonsorted CTLs, with no background lysis of HLA-A2.1+ normal cells. Cytoplasmic proteinase-3 expression was one log greater in CML blasts than in normal granulocytes. These results show that a PR1-HLA-A2 tetramer can be used to identify and select CTLs from normal donors that preferentially lyse CML cells, which could be used for leukemia-specific adoptive immunotherapy. PMID- 10363993 TI - Genetic alterations in bronchial lavage as a potential marker for individuals with a high risk of developing lung cancer. AB - Using 12 microsatellite markers, we have studied DNAs from the bronchial lavage of 90 individuals who were referred to an early-lung-cancer clinic in the Northwest of England with suspected lung cancer. Genetic alterations were detected in 15 (35%) of 43 patients with lung cancer but also in 11 (23%) of 47 patients with no cytological or radiological evidence of bronchial neoplasia. No significant differences were found between the referring symptoms in any of the second group of individuals with and without genetic alterations. No correlation was found between smoking exposure and loss of heterozygosity (LOH)/microsatellite alterations (MAs) in the microsatellite markers. On comparing LOH with MAs based on cytology review, we found that the prevalent type of alteration in specimens with cytological evidence of malignancy was LOH; in contrast, the individuals with no cytological evidence of malignancy showed a preponderance of MAs (P = 0.01). Our results indicate that a substantial proportion of cells in the bronchial lavage from suspected lung cancer patients carry identifiable genetic alterations. However, the presence of genetic alterations in the bronchial lavage of individuals with no clinical evidence of lung cancer raises the question whether instability is a phenomenon solely associated with cancer or represents a feature of nonneoplastic diseases. Our results suggest that microsatellite PCR-based assays can be developed as tools for the earlier identification of genetic changes in cells exfoliating in the bronchus. PMID- 10363992 TI - Reduced Fhit expression in sporadic and BRCA2-linked breast carcinomas. AB - Evidence for alteration of the FHIT gene in a significant fraction of breast carcinomas has been reported, in apparent concordance with loss of heterozygosity (LOH) at chromosome region 3p14.2 in breast cancer and benign proliferative breast disease. A significantly higher frequency of LOH at the FHIT locus was reported for BRCA2-/- tumors, possibly due to misrepaired double-strand breaks at this common fragile region. To determine whether such genomic alterations lead to Fhit inactivation, we have assessed the level of Fhit expression by immunohistochemical detection in sporadic tumors and cancers occurring in BRCA2 999del5 carriers. To determine whether Fhit inactivation may have prognostic significance, we have also assessed expression of breast cancer markers and clinical features in sporadic tumors relative to Fhit expression. Of 40 consecutive sporadic breast carcinomas studied for tumor markers, 50% showed reduced Fhit expression. In these sporadic cancers, loss of Fhit expression was not correlated significantly with the presence or absence of other tumor markers. In a study of 58 sporadic and 34 BRCA2 999del5 Icelandic invasive cancers, there was a significant association of LOH at 3p14.2 with reduced expression of Fhit (P = 0.001); also the lower expression of Fhit and higher LOH at 3p14.2 in BRCA2 999del5 tumors relative to sporadic cancers was significant (P = 0.002). Thus, genetic alteration at the fragile site within the FHIT gene leads to loss of Fhit protein in a significant fraction of sporadic breast cancers and a much larger fraction of familial breast cancers with an inherited BRCA2 mutation, consistent with the idea that loss of BRCA2 function affects stability of the FHIT/FRA3B locus. PMID- 10363994 TI - A mitotic spindle requirement for DNA damage-induced apoptosis in Chinese hamster ovary cells. AB - Promiscuously reactive electrophilic agents induce DNA and other cellular damage. DNA repair-defective cells, when compared with genetically matched, repair proficient parental cells, provide a means to distinguish cellular responses triggered by individual genetic lesions from other macromolecular damage. The Chinese hamster ovary (CHO) cell line EM9 is hypersensitive to the alkylating agent ethyl methanesulfonate (EMS) and is unable efficiently to repair DNA single strand breaks in contrast to parental AA8 cells. EM9 was used to examine how CHO cells couple unrepaired DNA strand breaks to loss of viability. Flow cytometry revealed that EMS-treated EM9 cells underwent prolonged cell cycle arrest in G2, followed by entry into mitosis, micronucleation, and apoptosis. EM9 cells synchronized in G1 prior to EMS treatment entered mitosis 24-36 h after release from synchrony, approximately 12 h after untreated control cells. Mitoses in EMS treated cells were abnormal, involving multipolar mitotic spindles and elongated and/or incompletely condensed chromosomes. The mitotic spindle poison nocodazole reduced DNA damage-induced apoptosis by >60%, whereas the frequency of micronucleation was similar in the presence or absence of nocodazole. Flow cytometry revealed that nocodazole-treated cells sustained a second round of DNA replication without intervening mitosis. These results demonstrate that nuclear fragmentation and inappropriate DNA replication are insufficient to trigger apoptosis following DNA strand breakage and demonstrate a requirement for mitotic spindle assembly for this process in CHO cells. PMID- 10363996 TI - Characterization of a novel receptor that maps near the natural killer gene complex: demonstration of carbohydrate binding and expression in hematopoietic cells. AB - A novel type II integral membrane protein has been identified in the course of screening for genes overexpressed in a mouse model of chronic myelogenous leukemia blast crisis. This new protein, designated NKCL, consists of a 210-amino acid polypeptide with a short, NH2-terminal cytoplasmic tail of 17 amino acids preceding a transmembrane domain and a COOH-terminal extracellular region. The COOH-terminal 132 amino acids bear typical features of the C-type animal lectin carbohydrate-recognition domain. The Nkcl gene is unique in that it maps just proximal to the region of the genome that encodes group V members of the C-type animal lectin family near the natural killer gene complex on mouse chromosome 6, but its protein product also has features of several group II C-type animal lectins. Most notably, it has a complete Ca2+-binding site 2, which forms part of the sugar-binding site in other members of the family, and binds mannose in a Ca2+-dependent manner. Moreover, its expression is not restricted to natural killer cells, as reported for the majority of group V lectins. Nkcl is expressed in pluripotent myeloid precursors, precursor and mature macrophages, and neutrophils. PMID- 10363995 TI - A new mechanism of acquisition of drug resistance by partial duplication of topoisomerase I. AB - Topoisomerase (topo)-I targeting antitumor agents are very effective in vivo against various human cancers. The indolocarbazole compound 6-N-formylamino-12,13 dihydro-1,11-dihydroxy-13-(beta-D-glucopyranosyl)- 5H-indolo[2,3-alpha]pyrrolo [3,4-c]carbazole-5,7(6H)-dione (NB-506) is a potent inhibitor of the religation step of topo I reaction, like camptothecin (CPT). We established a NB-506 resistant cell line from murine leukemia cell line P388. This resistant cell line, P388/F11, exhibited 73-fold higher resistance to NB-506 and 3.5-fold higher cross-resistance to CPT than the parental cell line. No induction of cleavable complex formations induced by NB-506 and CPT were detected by K-SDS precipitation assays in P388/F11 cells. Analysis of nuclear extracts from P388/F11 cells revealed that the relaxation activity of topo I was one-quarter of that of the parental cells, and that the activity was resistant to induction of DNA cleavage by these drugs. Furthermore, Western blot and Northern blot analyses showed the expression of an abnormal-sized 170-kDa topo I protein and its 6.0-kb transcript and the absence of the normal topo I protein and transcript in P388/F11 cells. Analyses of the structure of the abnormal topo I transcript by reverse transcription-PCR and direct sequencing methods revealed that a large portion of the gene from codon 21 to codon 609 was duplicated in its coding region. This internal duplication resulted in in-frame fusion and, thus, production of a partially duplicated protein of 1357 amino acids. Finally, we expressed and purified the recombinant P388/F11 topo I in a baculovirus system. P388/F11 topo I showed similar catalytic activity to wild-type topo I, but reduced sensitivities to NB-506 and CPT. These results show that the altered sensitivity of duplicated topo I is involved in the NB-506 resistance of P388/F11 cells and indicate a novel resistant mechanism which involves duplication of the topo I gene. PMID- 10363997 TI - Identification of a human anti-CD55 single-chain Fv by subtractive panning of a phage library using tumor and nontumor cell lines. AB - A large naive human single-chain (sc) Fv phage library was used to search for tumor-associated antigens by panning with a lung adenocarcinoma cell line, 1264, and counter-selecting with a nontumor bronchial epithelial cell line, BEAS-2B. After three rounds of subtractive panning, 239 of 673 clones analyzed bound selectively to 1264 tumor cells in a phage ELISA. Diversity analysis of these tumor-selective clones by BstNI fingerprinting and nucleotide sequencing revealed 14 distinct scFv fragments. Four clones bound selectively to 1264 over BEAS-2B cells when analyzed by a more discriminating flow cytometric assay using scFv. Moreover, these clones showed only limited cross-reactivity to several primary human cell lines. One clone, LU30, also cross-reacted strongly with the lung adenocarcinoma line, A549. The LU30 antigen was identified as decay-accelerating factor (CD55) by expression cloning from a 1264 cDNA library. The mean number of decay-accelerating factor molecules on the surface of 1264 and BEAS cells used for panning and counter-selection was estimated as 75,000 +/- 5,000 and 13,000 +/ 10,000, respectively. Thus, phage library panning combined with expression cloning permits identification of antibodies and their cognate antigens for proteins that are differentially expressed on the surface of distinct cell populations. PMID- 10363998 TI - Integrin alpha(v)beta3 promotes M21 melanoma growth in human skin by regulating tumor cell survival. AB - Growth and dissemination of malignant melanoma has a profound impact on our population, and little is known concerning the mechanisms controlling this disease in humans. Evidence is provided that integrin alpha(v)beta3 plays a critical role in M21 melanoma tumor survival within human skin by a mechanism independent of its known role in angiogenesis. Antagonists of alpha(v)beta3 blocked melanoma growth by inducing tumor apoptosis. Moreover, M21 melanoma cell interactions with denatured collagen, a known ligand for alpha(v)beta3, caused a 5-fold increase in the relative Bcl-2:Bax ratio, an event thought to promote cell survival. Importantly, denatured collagen colocalized with alpha(v)beta3 expressing melanoma cells in human tumor biopsies, suggesting that alpha(v)beta3 interaction with denatured collagen may play a critical role in melanoma tumor survival in vivo. PMID- 10363999 TI - Increased smad expression and activation are associated with apoptosis in normal and malignant prostate after castration. AB - Transforming growth factor (TGF)-beta1 is induced in the prostate after castration and has been implicated in apoptosis of epithelial cells during involution. TGF-beta1-mediated receptor activation induces phosphorylation of Smad2 and Smad3, which form complexes with Smad4, that translocate to the nucleus to regulate transcription of target genes. Smad6 and Smad7 antagonize the action of signal-transducing Smads. We have examined the immunohistochemical expression of different Smad molecules in the epithelium of rat ventral prostate before and after castration, in androgen-sensitive Dunning R3327 PAP prostatic tumor cells from untreated and castrated rats, and after treatment with estrogen. In the ventral prostate, a significant increase of phosphorylated Smad2 (P-Smad2) was observed after castration. In prostatic tumor cells we observed an increased expression of Smad2 and P-Smad2 after treatment. The levels of Smad3 and, in particular, Smad4 were enhanced in the normal ventral prostate, as well as in the tumors after castration. Interestingly, Smad6 and Smad7 expression was also up regulated in cells with increased Smad2 activation. The staining for Smad2, P Smad2, Smad3, Smad4, and Smad7 was nuclear in some cells and was present in areas with a large number of apoptotic cells identified by various morphological criteria, formation of apoptotic bodies and, in adjacent sections, by terminal deoxynucleotidyl transferase-mediated nick end labeling assay. Our results suggest that the signal transduction pathway for TGF-beta, leading to apoptosis, is activated in the normal prostate after castration and in the tumor model after castration, without or with estrogen treatment. PMID- 10364000 TI - Prostaglandin J2 and 15-deoxy-delta12,14-prostaglandin J2 induce proliferation of cyclooxygenase-depleted colorectal cancer cells. AB - Increased expression of cyclooxygenase (COX) and overproduction of prostaglandins (PGs) have been implicated in the development and progression of colorectal cancer (CRC). Nonsteroidal anti-inflammatory agents (NSAIDS) inhibit growth of various CRC cell lines by both COX-dependent and COX-independent pathways. To specifically examine the effect of COX and PGs on proliferation in CRC cells, we introduced an antisense COX-2 cDNA construct under the control of a tetracycline (Tc)-inducible promoter into a CRC cell line, HCA-7, Colony 29 (HCA-7) that expresses COX and produces PGs. In the presence of Tc, PG production in COX depleted cells was reduced 99.8% compared with either uninduced transfectants or parental HCA-7 cells. This decrease in PG production was associated with a concomitant 60% reduction in DNA replication. Subsequently, we examined the effects of various PGs to modulate cell growth in COX-depleted HCA-7 or COX-null HCT-15 cells by quantifying [3H]thymidine incorporation and/or growth in collagen gels. We report that J-series cyclopentenone PGs, particularly PGJ2 and 15-deoxy delta12,14-PGJ2, induce proliferation of these cells at nanomolar concentrations. Lipids extracted from parental HCA-7 cell conditioned medium stimulated mitogenesis in COX-depleted HCA-7 cells and COX-null HCT-15 cells. Using chromatographic and mass spectrometric approaches, we were able to detect PGJ2 in conditioned medium from parental HCA-7 cells. Taken together, these findings implicate a role for cyclopentenone PGs in CRC cell proliferation. PMID- 10364002 TI - What constitutes a total colonoscopy? PMID- 10364001 TI - Relation of TNF-related apoptosis-inducing ligand (TRAIL) receptor and FLICE inhibitory protein expression to TRAIL-induced apoptosis of melanoma. AB - Past studies have shown that apoptosis mediated by TNF-related apoptosis-inducing ligand (TRAIL) is regulated by the expression of two death receptors [TRAIL receptor 1 (TRAIL-R1) and TRAIL-R2] and two decoy receptors (TRAIL-R3 and TRAIL R4) that inhibit apoptosis. In previous studies, we have shown that TRAIL but not other members of the tumor necrosis factor family induce apoptosis in approximately two-thirds of melanoma cell lines. Here, we examined whether the expression of TRAIL-R at the mRNA and protein level in a panel of 28 melanoma cell lines and melanocytes correlated with their sensitivity to TRAIL-induced apoptosis. We report that at least three factors appear to underlie the variability in TRAIL-induced apoptosis. (a) Four of nine cell lines that were insensitive to TRAIL-induced apoptosis failed to express death receptors, and in two instances, lines were devoid of all TRAIL-Rs. Southern analysis suggested this was due to loss of the genes for the death receptors. (b) Despite the presence of mRNA for the TRAIL-R, some of the lines failed to express TRAIL-R protein on their surface. This was evident for TRAIL-R1 and more so for the TRAIL decoy receptors TRAIL-R3 and -R4. Studies on permeabilized cells revealed that the receptors were located within the cytoplasm and redistribution from the cytoplasm may represent a posttranslational control mechanism. (c) Surface expression of TRAIL-R1 and -R2 (but not TRAIL-R3 and -R4) showed an overall correlation with TRAIL-induced apoptosis. However, certain melanoma cell lines and clones were relatively resistant to TRAIL-induced apoptosis despite the absence of decoy receptors and moderate levels of TRAIL-R1 and -R2 expression. This may indicate the presence of inhibitors within the cells, but resistance to apoptosis could not be correlated with expression of the caspase inhibitor FLICE inhibitory protein. mRNA for another TRAIL receptor, osteoprotegerin, was expressed in 22 of the melanoma lines but not on melanocytes. Its role in induction of apoptosis remains to be studied. These results appear to have important implications for future clinical studies on TRAIL. PMID- 10364003 TI - Screening for hemochromatosis: phenotyping or genotyping or both? PMID- 10364004 TI - Updated guidelines for the diagnosis and treatment of gastroesophageal reflux disease. The Practice Parameters Committee of the American College of Gastroenterology. PMID- 10364005 TI - Saliva and esophageal protection. AB - There has been much interest in recent years in the potential protective role of saliva in the esophagus. Variables such as salivary volume and neutralizing capacity have been studied both during basal conditions and in response to esophageal acid exposure, in healthy subjects and in patients with esophagitis. In addition to its known neutralizing capacity, saliva also contains growth factors. These polypeptides (of which epidermal growth factor has been studied most) have cytoprotective and healing properties in various segments of the gastrointestinal tract. Therefore, a deficiency in one or more of these growth factors might be a contributing factor in the development of gastroesophageal reflux disease (GERD) or its complication, such as Barrett's metaplasia. However, human studies have produced contradictory results regarding salivary growth factor deficiency in such patients. Current methods of investigation make it difficult to assess the importance of saliva in GERD. This may be due in part to the multifactorial nature of the disease and the difficulty in long-term, selective manipulation of salivary function in humans. Given the present data in the literature, it is therefore unknown if saliva plays an important role in esophageal protection. PMID- 10364006 TI - Consultant strategies for the gastroenterologist. AB - Effective communication skills and the ability to respond to the needs of referring physicians are critical for the maintenance and enhancement of a referral base. One challenge is to determine the reason for the consultation and to respond to the referring physician's interests. The consultation letter should be formatted effectively and the "turn-around time" should be as brief as possible, perhaps by fax as well as mail. Prioritizing the differential diagnosis and diagnostic studies can be helpful to the referring physician. Strategies should be developed to address procedural referrals you may believe are inappropriate. A decision should be made as to whether the consultant or the primary care physician informs the patient of a serious disease diagnosis. Referral guidelines should be developed to assist primary care physicians to determine when a patient should be referred to the gastroenterologist or other specialist. Receptionists should avoid turning away new patients because of a "full schedule" and must determine if other arrangements can be made. A proper response to "hallway consultations" by referring physicians is important. Consultants should make efforts to increase their visibility in the medical community, either by newsletter or lectures to the medical staff. Every effort should be made to avoid speaking disparagingly about other physicians. Consultant strategies should be taught to house staff and fellows in a formal lecture format, in the outpatient clinics, and on hospital rounds. PMID- 10364007 TI - Esophageal solid bolus transit: studies using concurrent videofluoroscopy and manometry. AB - OBJECTIVE: Our aim was to assess the efficacy and mechanism of solid bolus transit through the esophagus. METHODS: Eight healthy volunteers were studied with concurrent manometry and videofluoroscopy while swallowing 5 ml liquid barium, a 5-6 mm diameter bread ball, and 4 g chewed bread in both a supine and upright posture. As many as four successive swallows were performed until clearance was achieved. RESULTS: The esophageal clearance of liquid barium was 100% with the first swallow. Clearance of the unchewed bread ball occurred with the first swallow in only 6.7% of trials in the upright posture and 5.9% in the supine posture. After four swallows, clearance was 100% and 52.9% in the upright and supine postures, respectively. Chewed bread was more readily cleared than unchewed bread, with 100% clearance after two swallows in the upright posture and 91% clearance after four swallows in the supine posture. The most common locus of bread stasis was at the aortic arch and carina. The bread boluses were noted to move more effectively when localized in the head as opposed to the tail of the bolus composite. Nonocclusive contractions often occurred at the bolus tail despite the increased peristaltic amplitude seen with the chewed bread. Failed peristalsis, a frequent cause for solid clearance failure, was observed during 30% of all bread swallows. This usually occurred distal to the stopping point of the bolus, suggesting it to be the result rather than the cause of impaired transit. CONCLUSIONS: Although infrequently perceived by these normal subjects and in contradistinction to liquid clearance, bread is rarely cleared from the esophagus with a single swallow. Mastication and an upright posture facilitate the esophageal transport of solids. Bolus composition and impaired bolus transit alter the amplitude and conductance of peristalsis. Manometric data pertaining to liquid clearance through the esophagus do not readily apply to bread. PMID- 10364008 TI - Ineffective esophageal motility: the most common motility abnormality in patients with GERD-associated respiratory symptoms. AB - OBJECTIVE: The association of gastroesophageal reflux disease (GERD) and respiratory symptoms is well known. The coexistence of ineffective esophageal motility (IEM, low-amplitude [< 30 mm Hg] or nontransmitted contractions in > or = 30% of 10 wet swallows in the distal esophagus) in patients with GERD has recently been demonstrated. Our aim was to determine the prevalence of IEM in patients with GERD-associated respiratory symptoms. METHODS: Manometry and pH studies of 98 consecutive patients with respiratory symptoms and abnormal reflux shown by pH-metry were reviewed. Symptoms were chronic cough (n = 43), asthma (n = 13), and laryngitis (n = 42). Sixty-six patients with heartburn with no extraesophageal manifestations were used as a control group. Total esophageal acid clearance (EAC) time was calculated for each patient. RESULTS: IEM was the most common motility abnormality seen in all groups of GERD patients. It was seen significantly more often in patients with chronic cough (41%) (p = 0.003) or asthma (53%) (p = 0.01), and numerically more often in patients with laryngitis (31%) than in patients with heartburn (19%). Diffuse esophageal spasm and nutcracker esophagus were rarely seen. Incidence of hypertensive or hypotensive lower esophageal sphincter was similar across all groups. The total EAC time was longer (median: 1.51 min/episode) (p = 0.01) in patients with GERD-associated respiratory symptoms than in patients with heartburn (median: 0.72 min/episode). CONCLUSIONS: IEM is the most prevalent motility abnormality in patients with GERD associated respiratory symptoms. Coexistence of IEM with GER may place patients at high risk for respiratory symptoms due to the associated delayed esophageal acid clearance seen with this motility abnormality. PMID- 10364009 TI - Does progesterone fluctuation across the menstrual cycle predispose to gastroesophageal reflux? AB - OBJECTIVE: Gastroesophageal reflux (GER) occurs in 30-50% of all pregnancies. The progressive rise in plasma progesterone has been suggested as a possible mediator of GER during pregnancy. It is not known whether progesterone, at physiological concentrations, has an effect on acid contact time. We sought to evaluate the relationship between progesterone concentrations, lower esophageal sphincter pressure (LESP), and acid contact time across the normal menstrual cycle. METHODS: LESP, 24-h ambulatory esophageal pH monitoring, and serum progesterone levels were determined in 19 healthy women known to have normal menstrual cycles. All tests were performed during the follicular phase (days 2-7) and the luteal phase (days 22-28) of one or two consecutive menstrual cycles. RESULTS: Despite marked oscillations in progesterone levels between the follicular phase (0.37 +/- 0.3 ng/ml) and luteal phase (4.64 +/- 2.92 ng/ml) we observed no significant differences in LESP (29.82 +/- 9.49 vs 30.45 +/- 8.56 mm Hg) or 24-h ambulatory pH levels (pH < 4) in total time (3.04 +/- 0.3% vs 3.18 +/- 2.51%), upright time (4.41 +/- 3.54% vs 4.18 +/- 3.36%), or supine time (0.77 +/- 1.32% vs 1.42 +/- 2.18%). CONCLUSIONS: The fluctuations in progesterone levels across the normal menstrual cycle have no significant impact on LESP and 24-h ambulatory pH parameters. Progesterone, at physiological concentrations, does not predispose to GER in healthy menstruating women. PMID- 10364010 TI - The cost-effectiveness of strategies to assess gastroesophageal reflux as an exacerbating factor in asthma. AB - OBJECTIVES: We sought to evaluate the cost-effectiveness of diagnostic strategies to determine whether or not acid reflux exacerbates asthma, and to identify which asthma response probabilities are most important in a cost-effective workup of this problem. METHODS: We performed a cost-effectiveness analysis, comparing 11 diagnostic strategies to assess the role that acid reflux plays in asthma. Probabilities and costs were derived from the published literature. Average and incremental costs, effectiveness, and cost-effectiveness were calculated for each strategy. Sensitivity analyses were performed. RESULTS: The most cost-effective diagnostic approach is to begin with omeprazole 20 mg/day for 3 months, followed by 24-h pH testing on drug in nonresponders. If 24-h pH testing is positive, increase the omeprazole dose every 3 months until the patient responds or a maximum of 60 mg/day is given. This strategy costs $730 per case correctly diagnosed. When the cost of pH testing exceeds $586 or the cost of omeprazole 20 mg/day is <$53 per month, omeprazole 20 mg/day for 3 months followed by 60 mg/day for the same duration in nonresponders becomes more cost-effective. CONCLUSIONS: Empiric acid reflux suppression, followed by pH testing in nonresponders, is the most cost-effective means of determining whether GERD is aggravating a patient's asthma. PMID- 10364011 TI - Motor and sensory function of the proximal stomach in reflux disease and after laparoscopic Nissen fundoplication. AB - OBJECTIVE: After Nissen fundoplication, dyspeptic symptoms such as fullness and early satiety develop in >30% of patients. These symptoms may result from alterations in proximal gastric motor and sensory function. METHODS: We have evaluated proximal gastric motor and sensory function using an electronic barostat in 12 patients after successful laparoscopic Nissen fundoplications (median follow-up; 12 months). Twelve age- and gender-matched patients with severe gastroesophageal reflux disease (GERD) and 12 healthy volunteers served as controls. Studies were performed in the fasting state and after meal ingestion. Gastric emptying tests were performed in all patients. Vagus nerve integrity was measured by the response of pancreatic polypeptide (PP) to insulin hypoglycemia. RESULTS: Minimal distending pressure and proximal gastric compliance were not significantly different between post-Nissen patients, GERD patients, and healthy controls. Postprandial relaxation of the stomach, however, was significantly (p < 0.05) reduced post-Nissen (267 +/- 34 ml), compared with controls (400 +/- 30 ml) and GERD (448 +/- 30 ml). Postprandial relaxation was significantly (p < 0.01) prolonged in GERD patients. Postprandial relaxation of the stomach correlated with gastric emptying of solids (r = 0.62; p = 0.01). Gastric emptying of solids became significantly (p < 0.05) faster after fundoplication. Postprandial fullness was significantly (p < 0.05) increased in the operated patients. CONCLUSIONS: Post-Nissen patients have a significantly reduced postprandial gastric relaxation and significantly accelerated gastric emptying, which may explain postoperative dyspeptic symptoms. The abnormalities result from fundoplication and not from vagus nerve injury or reflux per se, because in reflux patients gastric relaxation and gastric emptying are prolonged. PMID- 10364012 TI - How important is the route of reconstruction after esophagectomy: a prospective randomized study. AB - OBJECTIVE: A prospective randomized trial was performed to compare retrosternal and posterior mediastinal gastric tube reconstruction with regard to postoperative function and quality of life. METHODS: Twenty-six patients were randomly allocated to either retrosternal (n = 14) or posterior mediastinal (n = 12) reconstruction after gastric tube formation. Radionuclide transit studies were applied to obtain objective functional data and a standardized quality-of life assessment was performed. RESULTS: Retrosternal reconstruction showed an increased morbidity (15 vs 13 major complications) and mortality (14.2 vs 8.3%). Radionuclide clearance in the supine position was delayed in the gastric tube in general, compared with normal controls (retention index > 40% vs < 10%). There was a significantly higher retention (p < 0.005) in the retrosternal group in the middle third of the tube and the whole tube after intake of the liquid tracer. The retention of the first solid tracer was also higher in the retrosternal group in the middle third of the tube (p = n.s.) and was significantly higher in the whole tube after 30 (p < 0.05) and 60 (p < 0.01) s. This had no significant impact on the patients' quality of life. CONCLUSIONS: The posterior mediastinal route of reconstruction is recommended but curative resection (R0) is mandatory to avoid possible complications due to local tumor relapse. After incomplete resection (R1 or R2) we recommend retrosternal reconstruction for better palliation. PMID- 10364013 TI - Endoscopic dilation of benign esophageal strictures: report on 1043 procedures. AB - OBJECTIVE: Endoscopic dilation is considered the best treatment for most cases of benign esophageal stricture, although the best dilation technique and the kind of stricture is the most amenable to treatment is still controversial. We report on our experience on a large series of patients treated by dilation without the aid of fluoroscopy and compare the results of this therapy among patients with strictures from different causes. METHODS: Between 1992 and 1997, we performed 1043 dilation sessions on 153 patients. Treatment was considered adequate if the esophageal lumen could be dilated up to the size of a 42F catheter. If the stricture recurred after initial successful treatment, the stricture was dilated again up to a 42F catheter. RESULTS: One hundred forty patients (96 men, 44 women; mean age, 54.1 yr) were followed-up for a mean of 20.5 months (4 to 62 months). Stricture's etiology was postsurgical in 80 patients, peptic in 37, caustic in 12, and from other causes in 11 patients. Adequate dilation was achieved in 93.5% of the patients (131 of 140). Patients with peptic strictures needed a median of three sessions to be adequately dilated during follow-up in comparison to five sessions among patients with postsurgical or caustic strictures (p = 0.07). There were four perforations, with one death (2.8% and 0.7% per patient and 0.4% and 0.1% per session, respectively). CONCLUSIONS: Endoscopic dilation without the aid of fluoroscopy is safe and effective in relieving dysphagia caused by benign strictures of different causes, although repeated sessions are necessary because of stricture recurrence. PMID- 10364014 TI - Genotypes of Helicobacter pylori obtained from gastric ulcer patients taking or not taking NSAIDs. AB - OBJECTIVE: Whether Helicobacter pylori infection and use of nonsteroidal antiinflammatory drugs (NSAIDs) are independent risk factors for ulcerogenesis remains unclear. We undertook this study to evaluate H. pylori isolates from gastric ulcer patients to determine whether the genotype of the infecting isolate could be correlated with the use or nonuse of NSAIDs. METHODS: Fifty-two patients presenting with gastric ulcer and infected with H. pylori were included; 26 patients were taking NSAIDs or aspirin (ASA) regularly at the time of ulcer diagnosis. Polymerase chain reaction (PCR) was employed to assess the presence and mosaicism of the following H. pylori genes: cagA, vacA, iceA, and picB. RESULTS: We found no statistical differences in the presence of these genes in H. pylori isolates from gastric ulcer patients taking or not taking prescription NSAIDs or ASA. A 297-bp fragment of the cagA gene was detected in 96% of the isolates from the NSAID and ASA users and 100% from the non-NSAID users (p = 1.0). A larger and more variable region of the cagA gene was detected more frequently among the isolates from non-NSAID users than those from NSAID users (p = 0.05). Ninety-two percent of the isolates were identified as vacA genotype s1. The dominant vacA subtype was s1b, 76.9% and 65.4% in isolates from non-NSAID taking or NSAID-taking patients, respectively (p = 0.4). iceA1 genotype was not correlated with gastric ulcer as this allele was only detected in 17.3% of all isolates. CONCLUSIONS: No significant differences in the presence of the candidate virulence genes vacA, cagA, picB, or iceA were detected in isolates from gastric ulcer patients taking prescription NSAIDs or ASA, compared with those not taking these drugs, indicating that single gene presence does not allow discrimination of isolates that may be important in NSAID-induced ulcerogenesis. A variable region of the cagA gene was more frequently detected in isolates from patients not taking NSAIDs or ASA, suggesting that this gene may be modified by NSAID- or ASA-related factors or that certain strains may be selected for in patients taking these medications. PMID- 10364015 TI - Effects of Helicobacter pylori on proliferation of gastric epithelial cells in vitro. AB - OBJECTIVE: H. pylori infection of the gastric mucosa has been associated with an increase in gastric epithelial cell proliferation. However, in vitro adherence of H. pylori to gastric epithelial cells is associated with reduced cell proliferation. Reduction of epithelial cell proliferation may contribute to ulcer formation and delay ulcer healing. The following study was undertaken to elucidate the ability of cagA-positive and -negative strains to impede gastric epithelial cell proliferation. METHODS: A human gastric adenocarcinoma cell line (AGS) was overlaid with either cagA-positive or cagA-negative H. pylori strains suspended in cell culture medium. Proliferation of AGS cells was analyzed by performing direct cell counts and by measuring metabolism of a soluble tetrazolium compound (MTS), after exposure to H. pylori for 24 h. RESULTS: When compared with control cells cultured in medium alone, AGS cell proliferation was reduced by 45.6% and 28.5% due to exposure to cagA-negative and cagA-positive strains, respectively. When bacterial-induced cytotoxicity was assessed by measuring release of lactose dehydrogenase (LDH) into the culture medium, cagA positive strains were shown to induce significantly more cytotoxicity than cagA negative strains. CONCLUSIONS: These experiments demonstrate that H. pylori exposure to AGS cells significantly reduces cell proliferation. However, cagA positive strains that induce more cell injury reduce cell proliferation to a lesser extent than cagA-negative strains. Persistent replication of gastric epithelial cells injured by exposure to cagA-positive strains may be partially responsible for the stronger association with gastric cancer in persons infected with cagA-positive H. pylori strains. PMID- 10364016 TI - A comparison of three fingerstick, whole blood antibody tests for Helicobacter pylori infection: a United States, multicenter trial. AB - OBJECTIVE: We compared three whole blood antibody tests for Helicobacter pylori (H. pylori) in a United States, multicenter trial. METHODS: Patients referred for EGD at three medical centers were recruited. During EGD, biopsies were taken for histology and rapid urease testing (RUT). Immediately after endoscopy, patients underwent the antibody tests (FlexPack HP, Abbott Diagnostics; QuikVue, Quidel Corporation; AccuMeter, ChemTrak) using whole blood obtained by two to three fingersticks. Performance characteristics were calculated for each antibody test using the biopsy-based methods as a gold standard. RESULTS: A total of 131 patients participated; 50 (38%) patients had histological evidence of H. pylori infection. Using histology as a gold standard, the sensitivities of FlexPack HP, QuikVue, and Accumeter were 76%, 78%, and 84%, respectively. Specificity was 79% with FlexPack HP and 90% with QuikVue and Accumeter. There were no significant differences in the performance of the three antibody tests though there was a trend toward superior performance for AccuMeter compared to FlexPack HP (p = 0.019). However, RUT proved superior to FlexPack HP using histology as a gold standard (p = 0.008). Using either concordant histology and RUT results or a positive histology or RUT to define active H. pylori infection, there was no statistically significant difference between the antibody tests. CONCLUSIONS: There were no statistically significant differences in the performance of the three antibody tests. These tests proved only marginally sensitive in detecting patients infected with H. pylori. Clinicians should be aware of the limitations of these tests, particularly when using them as a sole means of testing for H. pylori. PMID- 10364017 TI - Relation between vacA subtypes, cytotoxin activity, and disease in Helicobacter pylori-infected patients from The Netherlands. AB - OBJECTIVE: The vacuolating cytotoxin of Helicobacter pylori (H. pylori) is encoded by vacA, of which allelic variation has been described. In the U.S., H. pylori strains with the signal sequence allele s1a are associated with enhanced gastric inflammation and with peptic ulcer disease (PUD). The m1 middle region allele is linked with more severe gastric epithelial damage. However, the distribution of H. pylori genotypes and the association with disease may be different in other geographical areas. The aim of this study was to establish whether vacA types among H. pylori isolates from Dutch patients are associated with disease. METHODS: The cytotoxin activity of the H. pylori isolates from 34 PUD patients and 46 patients with functional dyspepsia (FD) was assessed by an in vitro assay using HeLa cells as indicator cells. The vacA types and cagA status of the isolates were assessed by polymerase chain reaction (PCR). RESULTS: vacA s1-type H. pylori displayed cytotoxin activity more frequently than s2 vacA-type H. pylori (p = 0.003). This difference was not significant when only cagA+ H. pylori were considered. H. pylori isolates with the m1 vacA type exhibited a higher cytotoxin activity, independent of cagA (p = 0.006). Ninety-four percent (32/34) of the PUD patients and 74% (34/46) of the FD patients were infected with s1 vacA-type H. pylori (p = 0.04). When only cagA+ H. pylori were considered, s1 vacA type was not associated with disease. In addition, neither the s1a nor s1b subtypes correlated with disease. CONCLUSIONS: An association between vacA subtypes and disease could not be established in this patient population, due to the strong linkage between vacA s1 type and cagA. PMID- 10364018 TI - The 13C-urea blood test accurately detects active Helicobacter pylori infection: a United States, multicenter trial. AB - OBJECTIVES: Current nonendoscopic tests for Helicobacter pylori include antibody tests and the urea breath test. After the administration of 13C-urea, serum bicarbonate measurement can identify those infected with H. pylori. In this study, our aims were to determine the accuracy of the urea blood test, and to compare the accuracy of the urea blood test with that of rapid urease testing of gastric biopsies. METHODS: This was a multicenter trial conducted at five sites within the U.S. Patients scheduled for endoscopy were recruited. During endoscopy, biopsies were obtained from the gastric body and antrum for histology and rapid urease testing. Patients underwent the urea blood test, which required the ingestion of 125 mg of 13C-urea after endoscopy. Thirty minutes later, a 3-ml blood sample was obtained and later analyzed by mass spectrometry for 13C bicarbonate. Performance characteristics for the urea blood test were calculated using the endoscopic biopsy tests as a gold standard. RESULTS: One hundred and twenty-one patients (54 infected) were enrolled. The urea blood test yielded sensitivity of 89%, specificity of 96%, positive predictive value of 94%, negative predictive value of 91%, and accuracy of 93% using histology as a gold standard. There was no difference between results obtained with the urea blood test and rapid urease testing of gastric biopsies. CONCLUSIONS: The urea blood test accurately identified active H. pylori infection. The performance characteristics of this nonendoscopic test were similar to those of endoscopic rapid urease testing. PMID- 10364019 TI - Direct determination of Helicobacter pylori vacA genotypes and cagA gene in gastric biopsies and relationship to gastrointestinal diseases. AB - OBJECTIVE: Our aim was to detect Helicobacter pylori (H. pylori) from gastric biopsies of 248 patients using a novel, polymerase chain reaction (PCR)-based methodology, which simultaneously facilitates the determination of H. pylori vacA genotypes and cagA gene. METHODS: A simple methodology for sample preparation was established and PCR was performed with primer systems for the 16S rRNA, vacA, and cagA genes, thus circumventing the need to culture H. pylori and to extract DNA from biopsy samples. RESULTS: Infection with H. pylori was detected in 147 (59.3%) of 248 patients. The vacA signal sequence genotype s1 was present in 104 (81.3%) of 128 H. pylori-positive patients, and 24 (18.8%) patients had the genotype s2. The vacA middle region types m1 and m2 were detected in 46 (35.9%) and 79 (61.7%) patients, respectively. The combinations s1/m2 (43%) and s1/m1 (35.9%) were found more frequently than s2/m2 (18.8%). The cagA gene was detected in 75 (72.1%) of 104 H. pylori-positive biopsies with the vacA genotype s1. All 24 biopsies with the type s2 were cagA negative. Strains of the type vacA s1 were found in 97% of H. pylori-positive patients with peptic ulcer disease and were associated with the presence of the cagA gene, whereas 96% of the strains of the type vacA s2 were detected in patients who only had nonulcer dyspepsia. CONCLUSIONS: Using a novel PCR-based methodology, H. pylori 16S rRNA gene, vacA genotypes, and cagA gene can now be rapidly detected directly in gastric biopsies with high accuracy. These data demonstrate that infection with H. pylori strains of the vacA s1 genotype and the cagA gene are more likely to result in peptic ulcer disease. Determination of vacA genotypes and cagA gene may contribute to the potential clinical identification of patients at different levels of risk. PMID- 10364020 TI - Alteration in gastrointestinal and neurological function after electrical injury: a review of four cases. AB - OBJECTIVE: Individuals exposed to an electrical injury develop a variety of complications, several of which are recognized years after the initial electrical shock. Alteration in gastrointestinal and nervous system function has been described in these patients, yet the frequency and character of these abnormalities are poorly understood. We reviewed records of 40 individuals with a history of electrical injury to identify evidence of delayed onset of complications. METHODS: Forty consecutive patients with electrical shock injuries were monitored for up to 5 yr after their traumatic event using a comprehensive systems review. Of the eight patients who described an alteration in their gastrointestinal and neurological functions, four agreed to undergo further testing. Investigations included a flexible sigmoidoscopy, anorectal manometry, stool evaluation, serological and biochemical serum analysis, and a psychological examination. RESULTS: Each of the four patients described an increase in stool frequency and urgency. Anorectal manometry detected a reduction in threshold to rectal balloon distention and an abnormal anal sphincter control. Bowel function improved with meselamine. Psychiatric symptoms involving memory and concentration were observed in varying degrees. CONCLUSIONS: To our knowledge, these induced physiological and psychological changes after exposure to electrical shock injury have not yet previously been described. Our findings should encourage further clinical investigations to better anticipate, diagnose, and manage these and other as yet unrecognized delayed complications of electrical shock injury. PMID- 10364022 TI - Prospective analysis of complications 30 days after outpatient upper endoscopy. AB - OBJECTIVE: The aim of this study was to compare complication rates reported by patients 30 days after outpatient upper endoscopy with those discussed at our monthly morbidity and mortality conference. We also intended to establish which complications were reported most frequently 30 days after upper endoscopy, and which patients or procedures involved the highest risk. METHODS: Trained interviewers performed standardized telephone interviews on consecutive outpatients undergoing upper endoscopy over a 1-yr period. Patients were queried regarding potential events related to their upper endoscopy in the 30 days subsequent, including symptoms, emergency room (ER) and/or physician visits, and hospitalizations. The indications, findings, and therapies were reviewed from endoscopic reports. RESULTS: A total of 473 patients were contacted 30 days after outpatient upper endoscopy and agreed to participate in our study. The most common complications reported by patients at 30 days were sore throat (9.5%) and abdominal discomfort (5.3%). Twelve patients (2.5%) required an ER/physician visit and five patients (1.1%) required hospitalization. The minority of both ER/physician visits (16.7%) and hospitalizations (40%) were discussed at our monthly morbidity and mortality conferences. CONCLUSIONS: More complications were reported by patients 30 days after outpatient upper endoscopy than were discussed at our monthly morbidity and mortality conferences. The most frequent complications reported by patients were sore throat and abdominal pain. The minority of ER/physician visits and hospitalizations were discussed at our morbidity and mortality conferences. PMID- 10364021 TI - Fifteen years later: colonoscopic retroflexion revisited. AB - OBJECTIVE: Retroflex views of the rectal vault are included in the teaching of colonoscopic technique but are not pervasive in clinical practice. The utility of adding a retroflex maneuver at the end of colonoscopy has yet to be determined. The aim of this study was to evaluate the additional benefit of a retroflex view of the rectal vault at the completion of colonoscopic examination. METHODS: A prospective study of consecutive colonoscopies performed by a single physician was conducted. The rectal vault was first visually inspected upon withdrawal of the colonoscope. The endoscope was then readvanced into the rectum and retroflexed to view the vault. Endoscopic findings on both views were recorded along with demographic patient information. The six groups of findings sought on the two views were: retained stool, abnormal hemorrhoids, erosions/ulcerations, polyps, masses, and normal examinations. A determination on whether retroflex views influenced patient diagnosis was recorded by the endoscopist. RESULTS: There were 453 patients enrolled: 182 (40.2%) male and 271 (59.8%) female, consisting of 216 African-Americans, 232 Caucasians, and five Asians. The retroflex maneuver was performed successfully in 445 of 453 patients. In all but nine cases, the retroflex view did not produce additional information. The nine findings included three inflammatory pseudopolyps, five hyperplastic polyps, and one case of erosions/ulcerations. CONCLUSIONS: In the majority of cases, retroflexing the endoscope does not produce additional information compared with the thorough examination in straight view. The retroflex view may be of benefit if there is suspicion of pathology upon insertion or withdrawal of the colonoscope. PMID- 10364024 TI - Unfractioned heparin in the therapy of patients with highly active inflammatory bowel disease. AB - OBJECTIVE: Unfractioned heparin reportedly improves severe ulcerative colitis and Crohn's disease, but most of the few observations made have been published as abstracts. This prospective study evaluated whether heparin results in improvement of disease activity in patients with highly active, refractory ulcerative colitis or Crohn's disease. METHODS: Thirteen patients with ulcerative colitis and four patients with Crohn's disease received continuous intravenous heparin, aiming at a partial thromboplastin time of about 60 s for 2 wk. The following 6 wk, patients injected 12,500 units of heparin twice daily. All patients received sulphasalzine (1 g t.i.d.). Clinical and laboratory data were assessed weekly during the first month of treatment and every other week thereafter. RESULTS: A significant decline of clinical activity (p = 0.0059), C reactive protein (p = 0.0119), and erythrocyte sedimentation rate (p = 0.0096) was observed in the ulcerative colitis patients. In Crohn's disease clinical activity and laboratory values remained unchanged. Seven patients with ulcerative colitis but none of the Crohn's disease patients achieved complete remission after an average of 4 wk. In ulcerative colitis the histology (p = 0.0431) but not the endoscopic score (p = 0.1088) improved significantly. In one patient with ulcerative colitis, massive colonic bleeding was observed on day 11 of the study. CONCLUSIONS: These data are further evidence of a beneficial effect of unfractioned heparin in the therapy of patients with highly active ulcerative colitis. Because of possible serious bleeding, intravenous heparin should be administered in hospitalized patients only. PMID- 10364023 TI - High-level disinfection of gastrointestinal endoscopes: are current guidelines adequate? AB - OBJECTIVE: For a germicide to obtain a high level disinfection (HLD) claim, FDA requires demonstration of a 6-log reduction of mycobacterial inoculum under worst case conditions. The purpose of this study was to assess the adequacy of current guidelines for high level disinfection of GI endoscopes using alkaline glutaraldehyde in simulated-use testing. METHODS: Various gastrointestinal endoscopes were contaminated with Mycobacterium chelonae in 46 experiments. Quantitative cultures were obtained from each endoscope channel separately after each step: inoculation, standardized manual cleaning, immersion in 2% glutaraldehyde (Cidex) for 10, 20, or 45 min at room temperature, 70% isopropanol rinse, and drying. RESULTS: Manual cleaning alone achieved a 4-log reduction. After 10 min of glutaraldehyde exposure, but before alcohol rinse, two of 10 experiments failed to achieve a 6-log reduction. However, after alcohol rinse, all 10 experiments achieved HLD. After 20 min of glutaraldehyde exposure, but before alcohol rinse, one of 18 experiments failed to achieve a 6-log reduction. After alcohol rinse, all 18 experiments achieved HLD. After 45 min of glutaraldehyde exposure, but before alcohol rinse, one of 18 experiments failed to achieve a 6-log reduction. After alcohol rinse, all 18 experiments achieved HLD. Thus, if the entire reprocessing protocol including manual cleaning, glutaraldehyde exposure, alcohol rinse, and drying was taken into account, the required 6-log reduction of mycobacteria was achieved with a minimum of 10 min of glutaraldehyde exposure at room temperature. CONCLUSIONS: Current guidelines for high level disinfection using glutaraldehyde are appropriate. Alcohol rinse is a valuable adjunctive step for drying and for its bactericidal effects. PMID- 10364025 TI - Fecal incontinence with normal anal canal pressures: where is the pitfall? AB - OBJECTIVE: One third of subjects who suffer from fecal incontinence are found to have values within the normal range when anal manometry is performed. For these patients, one hypothesis is that impaired rectal adaptation to distension may occur. The aim of our study was to analyze anorectal responses to rectal isobaric distension in this population. METHODS: This was a prospective study conducted in 51 consecutive incontinent patients (45 female, six male) divided into two groups according to their functional anal state: absence (19 patients aged 55 +/- 6 yr) or presence of manometric anal weakness (32 patients aged 59 +/- 2 yr). The subjects were submitted to two randomized modes of rectal isobaric distension (tonic, phasic) with an electronic barostat. Anal pressures, perception, and volumes of the rectum were recorded at six different preselected pressures. RESULTS: As compared with those having anal weakness, patients with no anal weakness retained higher mean pressures at both upper (36.9 +/- 2.2 vs 22.9 +/- 1.4 mm Hg; p = 0.01) and lower parts (41.0 +/- 2.0 vs 23.3 +/- 1.4 mm Hg; p = 0.002) of the anal canal, similar perception scores, but much lower rectal volumes (68.5 +/- 5.5 vs 121.8 +/- 7.0 ml; p = 0.008) in response to rectal isobaric distension. CONCLUSION: A decrease in rectal adaptation could be involved in fecal leakage in patients with no anal manometric weakness. PMID- 10364026 TI - Change in the extent of colonoscopic and histological involvement in ulcerative colitis over time. AB - OBJECTIVE: Colonoscopy has replaced barium enema as the method for determining the extent of disease in patients with ulcerative colitis (UC). Normally, the extent of disease is determined by direct visualization of the mucosa, but biopsies are also used with increasing frequency. Very little is known about the extent to which these two ways of assessing the extent of disease are correlated and whether the correlation differs over time. The aim of this study was to determine the changes in extent of disease assessed by direct visualization and by histological examination of the mucosa at the time of diagnosis and after 1 yr of follow-up in a cohort of incident cases of UC patients. METHODS: All new cases of UC in a defined population were identified during a 4-yr period (496 patients). Of these, 384 patients (78%) were available for follow-up and were subjected to a second colonoscopy with representative biopsies taken from both normal and affected mucosa. RESULTS: After 1 yr there were macroscopical signs of progression in 14%; 22% showed regression, and 30% had a normal colonoscopy. The histological changes from diagnosis until follow-up showed progression in 20%, 24% showed regression, and 24% had normal histological findings. Histological examination showed more extensive disease than did direct visualization in 4% of patients at diagnosis and in 28% at follow-up, whereas direct visualization showed more extensive disease than did histological examination in 18% of patients at diagnosis and 12% at follow-up. The best correlation at both diagnosis and follow-up was seen in pancolitis (99% and 88%, respectively). CONCLUSIONS: With regard to the extent of colonic involvement in the UC patients, we found less agreement between endoscopic and histological evaluation at the follow-up examination than at the start of the study. This could indicate that biopsies represent a better indicator than endoscopical examination for long term prognosis. Further studies are needed to confirm this finding. PMID- 10364027 TI - Gastrointestinal symptoms in long-distance runners, cyclists, and triathletes: prevalence, medication, and etiology. AB - OBJECTIVE: The aim of this study was to determine the prevalence of exercise related gastrointestinal (GI) symptoms and the use of medication for these symptoms among long-distance runners, cyclists, and triathletes, and to determine the relationship of different variables to GI symptoms. METHODS: A mail questionnaire covering the preceding 12 months was sent to 606 well-trained endurance type athletes: 199 runners (114 men and 85 women), 197 cyclists (98 men and 99 women), and 210 triathletes (110 men and 100 women) and sent back by 93%, 88%, and 71% of these groups, respectively. Symptoms were evaluated with respect to the upper (nausea, vomiting, belching, heartburn, chest pain) or lower part of the GI tract (bloating, GI cramps, side ache, urge to defecate, defecation, diarrhea). For statistical analysis, Mann-Whitney U test, Fisher exact test, or Student t test were used. RESULTS: Runners experienced more lower (prevalence 71%) than upper (36%) GI symptoms during exercise. Cyclists experienced both upper (67%) and lower (64%) symptoms. Triathletes experienced during cycling both upper (52%) and lower (45%) symptoms, and during running more lower (79%) than upper (54%) symptoms. Bloating, diarrhea, and flatulence occurred more at rest than during exercise among all subjects. In general, exercise-related GI symptoms were significantly related to the occurrence of GI symptoms during nonexercise periods, age, gender, diet, and years of training. The prevalence of medication for exercise-related GI symptoms was 5%, 6%, and 3% for runners, cyclists, and triathletes, respectively. CONCLUSIONS: Long-distance running is mainly associated with lower GI symptoms, whereas cycling is associated with both upper and lower symptoms. Triathletes confirm this pattern during cycling and running. The prevalence of medication for exercise-related GI symptoms is lower in the Netherlands in comparison with other countries, in which a prevalence of 10-18% was reported. More research on the possible predisposition of athletes for GI symptoms during exercise is needed. PMID- 10364028 TI - Epstein-Barr virus infection of the colon with inflammatory bowel disease. AB - OBJECTIVE: Epstein-Barr virus (EBV)-infected cells can evoke severe host immune responses, as shown in infectious mononucleosis and EBV-associated gastric carcinoma. To investigate the possible pathological role of EBV in inflammatory bowel disease (IBD), we tested for the presence of EBV in the colon in IBD patients. METHODS: Surgically resected colonic specimens of 11 patients with Crohn's disease, five patients with ulcerative colitis, nine noninflammatory controls (disease-free area of the colorectal carcinoma), and 10 appendicitis cases were tested using highly sensitive in situ hybridization for EBV-encoded small RNA1 (EBER-1). RESULTS: EBER-1 was detected in 63.6% of Crohn's disease cases and 60% of ulcerative colitis cases, but not at all in noninflammatory controls and appendicitis cases. EBER-1-positive cells were very rare in the noninflammatory areas of colonic specimens from IBD patients. EBER-1-positive cells were nonepithelial cells (mainly B lymphocytes and a few histiocyte-shaped cells) located in erosive or ulcerative areas of the colonic specimens. CONCLUSION: The limited presence of EBV-infected cells in the diseased areas of IBD colonic specimens indicated that EBV infection may be related to such diseases. PMID- 10364029 TI - Intravenous cyclosporin in ulcerative colitis: a five-year experience. AB - OBJECTIVE: Cyclosporin (CSA) is a promising alternative for patients with severe steroid-refractory ulcerative colitis (UC) previously facing only surgical options. Concerns over the long term efficacy and side effects resulted in this investigation of the University of Chicago's 5-yr CSA experience in these patients. METHODS: All steroid-refractory severe ulcerative colitis (UC) patients treated with IV CSA from 1991 to 1995 were identified by using the university's IBD database, with additional information from patient charts and physician files. RESULTS: A total of 42 patients with severe UC unresponsive to IV steroids were treated with IV CSA (4 mg/kg/day). Of 42 patients, 36 (86%) responded; 31 were continued on oral CSA (8 mg/kg/day) for an overall mean of 20 wk. Ten initial CSA responders had colectomies after a mean of 6 months. Of the 36 initial responders, 25 (69%) also received 6-mercaptopurine (6-MP) or azathioprine (aza), and CSA and steroids were tapered. A total of 20% required colectomy, vs 45% of those not receiving 6MP/aza. In all, 62% of all patients, 72% of initial CSA responders, and 80% of initial CSA responders receiving 6MP/aza have avoided colectomy, with a life table analysis of "noncolectomy survival" of 58%, 70%, and 71%, respectively, at 5.5 yr. All colectomies occurred within 18 months of CSA initiation. Complications, resulting in CSA discontinuation in six patients, were all reversible, with complete recovery. CONCLUSIONS: CSA successfully allows most severe steroid resistant UC patients to retain their colons, and provides time for "elective" colectomy in others, especially if 6MP/aza are also given. Careful monitoring for side effects, including PCP prophylaxis, should be part of the treatment protocol. PMID- 10364030 TI - Screening blood donors for hereditary hemochromatosis: decision analysis model comparing genotyping to phenotyping. AB - OBJECTIVE: The identification of a gene for hereditary hemochromatosis in 69-100% of typical hemochromatosis patients has resulted in a genotypic test to identify persons with the typical missense mutation. Population screening by genotyping has the potential to reduce screening costs because of a high specificity of the genetic test. METHODS: Decision analysis techniques are used to compare the outcome, utility, and incremental cost savings of a plan to screen voluntary blood donors and their siblings for hemochromatosis using a genotypic test (C282Y mutation) with phenotypic tests (transferrin saturation, serum ferritin). RESULTS: Genotypic screening is less expensive than phenotypic screening only if the cost of the initial genetic test is less than $20. The screening program saves money (dominant strategy) if the cost of the initial genetic test is less than $28. Incremental cost saving declines as the cost of the gene test increases. At a gene test cost of $173, it costs $109,358 to identify a homozygote with potential life-threatening illness. Incremental cost saving also declines as the penetrance of the hemochromatosis gene in the population screened decreases. Phenotypic screening with confirmatory genetic testing results in a cost of $2,711 per homozygote with life-threatening complications. CONCLUSIONS: Population screening programs for hemochromatosis have the potential to save money. Optimal strategies for screening include initial testing for iron overload (phenotyping) with confirmatory genetic testing, or initial genetic testing if the test is less than $28. PMID- 10364031 TI - Immunogenicity of hepatitis A vaccine in decompensated liver disease. AB - OBJECTIVE: Hepatitis A can cause decompensation and death in patients with previous liver injury. The hepatitis A vaccine is recommended for patients with chronic liver disease. The aim of this study was to screen, immunize, and measure the safety and antibody response of the hepatitis A vaccine in liver failure and liver transplant patients. METHODS: This was a prospective immunization trial at a referral center for liver disease and liver transplantation. A total of 193 patients with severe chronic liver disease were screened and 24 patients were vaccinated. Sixteen end stage liver disease patients were compared with eight liver transplant patients. Hepatitis A vaccinations using 1440 ELISA units were given at 0 and 2 months. Serum hepatitis A antibody titers were measured after each vaccine dose. An antibody response > or = 33 mIU/ml was considered protective. RESULTS: Screening seropositive rate was 70 of 193 (36%) and 24 patients were available for vaccination. The median antibody titer was markedly lower in liver transplant patients, 0.0 mIU/ml compared to liver failure patients 34.7 mIU/ml (p < 0.001). Liver transplant recipients did not respond to the vaccine (0 of eight patients) compared with seven of 14 liver failure patients (seroconversion rate 50%, p = 0.02). CONCLUSIONS: Liver failure significantly reduces the antibody response to hepatitis A vaccine, and liver transplant recipients were unable to respond to the vaccine. Although this study was small, immunization should be considered early for susceptible patients with chronic liver disease because the development of liver failure may blunt the immunogenicity of the vaccine. PMID- 10364032 TI - A predictive model for the development of hepatocellular carcinoma, liver failure, or liver transplantation for patients presenting to clinic with chronic hepatitis C. AB - OBJECTIVE: Chronic infection with hepatitis C may lead to the development of cirrhosis, liver failure, and hepatocellular carcinoma. However, not all patients progress to these endpoints. Ideally, clinicians could improve their capability of stratifying the risk and the time frame within which their patients will progress to these endpoints. The purpose of the present study was to construct statistical models predicting disease progression for individual patients. METHODS: Study endpoints were the development of hepatocellular carcinoma, liver transplantation, or death due to liver disease. The study cohort was 256 patients with hepatitis C acquired from either blood transfusion or use of intravenous drugs. During follow-up, 17 patients developed hepatocellular carcinoma, seven received liver transplantation, and 12 died from liver disease. RESULTS: On multivariate analysis a history of decompensation (relative risk [RR] 4.321, 95% confidence interval [CI] 1.777-10.511) and the serum albumin (RR 0.253, 95% CI 0.136-0.474) were independently associated with the study endpoints. Patients without a history of decompensation and with a serum albumin > or = 4.1 mg/dl had a 3.2% chance of developing the study endpoints within 5 yr. Patients with a history of decompensation and a serum albumin < 4.1 mg/dl had a 40% chance of developing a study endpoint within 5 yr. Baseline genotype and quantitative RNA were not associated with development of the clinical endpoints, with the exception of patients coinfected with two or more genotypes. CONCLUSION: Thus, the serum albumin and a history of decompensation are useful for predicting the development of hepatocellular carcinoma, liver transplantation, and death due to liver disease among patients with hepatitis C. PMID- 10364033 TI - Cancer antigen 125: a sensitive marker of ascites in patients with liver cirrhosis. AB - OBJECTIVE: Cancer antigen 125 (CA 125) is a high molecular mass glycoprotein, usually used for monitoring the course of epithelial ovarian cancer. Recently it has been shown that liver cirrhosis is associated with increased levels of CA 125, particularly in the presence of ascites. The aim of this study was to evaluate CA 125 as a marker for the detection of ascites in patients with chronic liver disease. METHODS: A total of 170 patients were studied. All had ultrasound scanning for detection of ascites. Group I consisted of 123 patients with chronic liver disease without ascites; whereas group II consisted of 47 patients with chronic liver disease with ascites. CA 125 levels were measured in all patients and also simultaneously in the ascitic fluid of 31 patients from group II. RESULTS: Of 47 patients, 46 (97.8%) of group II had elevated serum levels of CA 125 (mean 321 +/- 283 U/ml) as compared with only nine of 123 (7.3%) patients of group I [mean 13 +/- 15 U/ml]), p < 0.001. The mean CA 125 concentration in the ascitic fluid of 31 cirrhotic patients (group II) was 624 +/- 397 U/ml and was always higher than corresponding serum levels (p < 0.01). Serum CA 125 levels correlated with the amount of ascitic fluid (r = 0.78). A profound decrease in serum CA 125 concentration was noted 2-3 and 10 days after large volume paracentesis. CA 125 was more sensitive and preceded ultrasonography in detection of ascites in few cirrhotic patients. CONCLUSIONS: CA 125 is a highly sensitive marker to detect ascites in patients with liver cirrhosis. This marker may be useful to detect small to moderate amounts of ascitic fluid in cirrhotic patients when physical examination is difficult or equivocal for ascites. PMID- 10364034 TI - Hepatitis C virus genotypes and viremia and hepatocellular carcinoma in the United States. AB - OBJECTIVE: Hepatitis C virus (HCV) is a well recognized cause of hepatocellular carcinoma (HCC). The pathogenic significance of HCV genotypes in hepatocarcinogenesis is undefined. The aim of this study was to investigate the genotypic distribution and viremic level of HCV in patients with HCV-associated cirrhosis with or without HCC. METHODS: A total of 28 HCV-infected patients with HCC (HCC+) and 38 patients with HCV-associated cirrhosis without HCC (HCC-) were studied. HCV genotype was assessed by the genotype-specific polymerase chain reaction (PCR) method of Okamoto and restriction fragment length polymorphism (RFLP) of the 5' untranslated region (5' UTR). Hepatitis C viremia was quantitated with the branched-chain DNA (bDNA) assay. RESULTS: Using the Okamoto method, we found genotype 1b in 64% of the HCC+ group and 74% of the HCC- group, 36% of the HCC+ group and 16% of the HCC- group were coinfected with a combination of genotype 1b and another genotype. Using the RFLP method, we found genotype 1b in 41% of the HCC+ group and in 24% of the HCC- group. Other genotypes accounted for 18% of the HCC+ group and 55% of the HCC- group; no combination genotypes were identified. Poor concordance occurred between the two genotyping methods. Mean bDNA levels were not significantly different between the two groups. CONCLUSIONS: Our study demonstrates that no particular HCV genotypes were associated with HCC and genotype did not appear to influence the development of HCV-associated HCC. PMID- 10364035 TI - Intraspousal transmission of GB virus C/hepatitis G virus in an hepatitis C virus hyperendemic area in Japan. AB - OBJECTIVE: An immunoassay for antibodies against an hepatitis G virus (HGV) protein (anti-E2) was recently developed that might serve as a useful marker for diagnosing recovery from HGV infection. METHODS: We investigated the intraspousal transmission of GB virus C/hepatitis G virus (GBV-C/HGV) using both reverse transcription hemipolymerase chain reaction (RT-hemi-PCR for the 5' untranslated region) and a recently developed anti-E2. RESULTS: Thirty-two GBV-C/HGV-infected index subjects were selected from an hepatitis C virus hyperendemic area in Japan. Of the 32 subjects, seven (6.4%) were GBV-C/HGV RNA-positive, 24 (21.8%) were anti-E2-positive, and one (0.9%) was both GBV-C/HGV RNA- and anti-E2 positive. Among the 32 spouses of these subjects, GBV-C/HGV RNA, anti-E2, and both GBV-C/HGV RNA and anti-E2 positivity were detected in 0, 6, (18.8%), and one (3.1%) spouses, respectively (the total prevalence of GBV-C/HGV was 7 spouses [21.9%]). Thus, the intraspousal transmission of GBV-C/HGV was undeniable in these seven couples. The respective positive rates of 175 sex- and age-matched controls were 7 (4.0%), 26 (14.9%), and 0 (the total prevalence of GBV-C/HGV was 34 [19.4%]). No significant difference in positive rates was observed between the subjects/spouses and the controls. Five spouses among the seven couples who were positive for any of GBV-C/HGV markers had parenteral risk factors such as blood transfusion, acupuncture, and major surgery. CONCLUSION: Based on these observations, we cannot draw a definitive conclusion that intraspousal transmission of GBV-C/HGV had occurred among these seven couples. PMID- 10364036 TI - Systemic hemodynamic changes in mansonic schistosomiasis with portal hypertension treated by azygoportal disconnection and splenectomy. AB - OBJECTIVE: The aim of this study was to assess systemic hemodynamic changes in patients with Manson's schistosomiasis and portal hypertension during azygoportal disconnection and splenectomy. METHODS: Sixteen patients with portal hypertension secondary to hepatosplenic schistosomiasis with indication for surgery were studied prospectively. All underwent invasive hemodynamic monitoring with pulmonary artery catheter. The first systemic hemodynamic assessment was performed preoperatively. In the intraoperative period new hemodynamic data were collected as follows: a) after laparotomy; b) 15-30 min after splenic artery ligature; c) 15-30 min after splenectomy; and d) after ligation of the collateral circulation. RESULTS: The results indicated preoperatively that the patients presented with an increased cardiac index (4.40 +/- 0.94 L/min/m2) together with a reduction in the systemic vascular resistance index (1692.25 +/- 434.91 dyne.s/cm5.m2). The stroke index (53.74 +/- 10.40 ml/beat/m2) and both left (5.71 +/- 1.50 kg.m/m2) and right heart work indexes (1.12 +/- 0.74 kg.m/m2) were also elevated. The mean pulmonary artery pressure was increased (17.81 +/- 9.00 mm Hg) and the pulmonary vascular resistance index decreased (164.31 +/- 138.69 dyne.s/cm5.m2). From the moment that the splenic artery was ligated until the end of the procedure, the cardiac index (3.45 +/- 0.90 L/min/m2) was reduced and the systemic vascular resistance index (2059.50 +/- 590.05 dyne.s/cm5.m5) increased. The systolic index (44.25 +/- 11.01 ml/beat/m2) and the left ventricle work index (4.33 +/- 1.29 kg.m/m2) also reduced. The mean pulmonary artery pressure (19.18 +/- 9.21 mm Hg) and the right ventricle work index (0.94 +/- 0.62 mm Hg) remained elevated after the surgical procedure. CONCLUSIONS: The data allowed us to conclude that hepatosplenic schistosomiasis induces a hyperdynamic circulatory state that was corrected after splenectomy and azygoportal disconnection, remaining a mild pulmonary hypertension. Therefore, these changes are correlated with the portosystemic collateral circulation, especially as a consequence of splanchnic hyperflow. PMID- 10364037 TI - Gene mutations of K-ras in gallbladder mucosae and gallbladder carcinoma with an anomalous junction of the pancreaticobiliary duct. AB - OBJECTIVE: In this study, we examined the mutational spectrum of K-ras in cases of gallbladder and gallbladder carcinoma with an anomalous junction of the pancreaticobiliary duct (AJPBD). METHODS: We examined 35 gallbladders with AJPBD (20 with hyperplasia, 15 with carcinoma) and 38 gallbladders without AJPBD (four normal gallbladders, four with hyperplasia, six with adenoma, 24 with carcinoma). Polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) and direct sequencing were performed to detect mutations in codon 12 or 13 of K-ras. RESULTS: In the cases with AJPBD, the prevalences of K-ras mutation were 15% (3/20) in hyperplasia, 60% (6/10) in stage I carcinoma, and 100% (5/5) in stage II-IV carcinoma. In the cases without AJPBD, the prevalences of K-ras mutation were 0% (0/4) in normal gallbladder, 0% (0/4) in hyperplasia, 17% (1/6) in adenoma, 7% (1/16) in stage I carcinoma, and 38% (3/8) in stage II-IV carcinoma. Prevalences of K-ras mutation in hyperplasia and carcinoma with AJPBD were greater than those without AJPBD (p < 0.05). The point mutation of GGT to GAT in codon 12 was frequently observed in the cases with AJPBD. CONCLUSION: These results suggest that the specific K-ras mutation in codon 12 (GGT to GAT) may contribute to the early stage of carcinogenesis in the gallbladder with AJPBD. PMID- 10364038 TI - The risk for cancer or dysplasia in ulcerative colitis patients with primary sclerosing cholangitis. AB - OBJECTIVES: Recent studies have implicated primary sclerosing cholangitis (PSC) as a risk factor for colorectal cancer (CRC) in ulcerative colitis (UC). Our study was designed to define both the risk and the risk factors for CRC or dysplasia in a large UC cohort with PSC. METHODS: Patients with UC and PSC were compared with a random sample of UC controls without PSC. Patients were analyzed from the inception of disease until an outcome or censor. RESULTS: Thirty-three (25%) of 132 UC patients with PSC developed CRC or dysplasia compared with 11 (5.6%) of 196 controls (adjusted relative risk 3.15, 95% confidence interval 1.37 7.27). Possible risk factors were chronic disease activity and lack of folate supplementation. Of 17 CRCs in the PSC group, 76% occurred proximal to the splenic flexure and 35% presented at an advanced stage, compared with one of five (20%) CRCs in controls being proximal and none being advanced. Six (4.5%) PSC patients, and no controls, died of CRC (p < 0.01). CONCLUSIONS: UC patients with PSC are at increased risk of developing CRC or dysplasia. Chronically active disease may be a risk factor, whereas folate could have a protective effect. CRCs associated with PSC are more likely to be proximal, to be diagnosed at a more advanced stage, and to be fatal. PMID- 10364039 TI - Utility valuations for outcome states of colorectal cancer. AB - OBJECTIVE: Utilities for the outcome states of colorectal cancer (CRC) must be measured to evaluate the cost-utility of screening and surveillance strategies for this disease. We sought to measure utilities for stage-dependent outcome states of CRC. METHODS: We identified persons who had previously undergone removal of colorectal adenoma. We conducted individual interviews in which these participants were presented with stage-dependent outcome states and were asked to assess utilities for them using the standard gamble technique. RESULTS: A total of 90 participants were interviewed; nine were excluded, leaving 81 for analysis. We obtained the following utility valuations: stage I rectal or stage I/II colon cancer (mean 0.74, median 0.75); stage III colon cancer (mean 0.67, median 0.75); stage II/III rectal cancer without ostomy (mean 0.59, median 0.60), stage II/III rectal cancer with ostomy (mean 0.50, median 0.55), stage IV rectal or colon cancer (mean 0.25, median 0.20). These valuations were statistically different from each other. CONCLUSIONS: We measured utilities for stage-dependent outcome states of CRC in a sample of persons who had previously undergone removal of colorectal adenoma. We found that our participants were able to differentiate between the presented outcome states and assigned lower utility to increasingly morbid states. Our results show that stage-dependent morbidity is an important consideration in CRC and should be incorporated into any decision analysis model evaluating the cost-effectiveness of CRC screening or surveillance. PMID- 10364040 TI - Alpha-fetoprotein-producing gastric cancer: histochemical analysis of cell proliferation, apoptosis, and angiogenesis. AB - OBJECTIVE: Alpha-fetoprotein (AFP)-producing gastric cancer has been associated with a poor prognosis. In the present study, the cell proliferation, apoptosis, and angiogenesis of this cancer were studied histochemically to determine its malignant potential. METHODS: Tissue samples were taken from four patients with AFP-producing gastric cancer and 26 patients with AFP-negative gastric cancer. Cell proliferation was evaluated by Ki-67 immunostaining, and the Ki-67 labeling index (LI) was determined. Apoptosis was studied by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling method, and the apoptotic index was determined. Angiogenesis was evaluated by measuring the microvessel density using factor VIII immunostaining, and immunostainings for vascular endothelial growth factor and thymidine phosphorylase were performed. RESULTS: The Ki-67 LI of the AFP-producing gastric cancers was significantly higher than that of the AFP-negative gastric cancers (p < 0.01). The apoptotic index of the AFP-producing gastric cancers was significantly lower than that of the AFP-negative gastric cancers (p < 0.01). The microvessel density of the AFP producing gastric cancers was significantly higher than that of the AFP-negative gastric cancers (p < 0.01). Vascular endothelial growth factor expression was observed in all four of the AFP-producing gastric cancers, whereas thymidine phosphorylase was not expressed in any of the AFP-producing gastric cancers. CONCLUSIONS: These results suggest that AFP-producing gastric cancers have high malignant potential (high proliferative activity, weak apoptosis, and rich neovascularization) compared with that of AFP-negative gastric cancers. These biological characteristics of AFP-producing gastric cancer reflect the aggressive behavior and the poor prognosis of patients with this type of cancer. PMID- 10364041 TI - Complete regression of recurrent esophageal carcinoma with reduced expression of glutathione S-transferase-pi by treatment with continuous infusion of 5 fluorouracil and low-dose cisplatin infusion. AB - The mortality rate of recurrent esophageal carcinoma remains high because of its resistance to chemotherapy and radiation therapy. We present a patient with recurrent esophageal carcinoma, which dramatically disappeared after treatment with the combination of continuous infusion of 5-fluorouracil and low-dose cis Diamminedichloroplatinum-II (cisplatin) infusion (FP therapy). Furthermore, we immunohistologically found that glutathione S-transferases (GST)-pi, a marker of resistance to cisplatin, was faintly expressed both in the endoscopical biopsy specimens of recurrent tumor and in the resected specimens of esophageal carcinoma and metastatic lymph nodes. FP therapy was suggested to be effective for recurrent esophageal carcinoma. Immunostaining for GST-pi might be a prospective marker for the sensitivity of esophageal carcinoma to FP therapy, particularly cisplatin. PMID- 10364042 TI - Primary hepatic lymphoma associated with primary biliary cirrhosis. AB - We report a case of primary hepatic lymphoma in a 55-yr-old female patient with primary biliary cirrhosis and Sjogren's syndrome. On July 1994, a tumor measuring 11 mm in diameter was detected in the right lobe of the liver by abdominal ultrasonography. A needle biopsy specimen showed the lesion to contain small- and medium-sized lymphoid cells without obvious atypia, and a provisional diagnosis of pseudolymphoma was made. About 2 yr later, the tumor increased to 15 mm in diameter, necessitating a second needle biopsy. Histological and genetic examinations confirmed non-Hodgkin's lymphoma of diffuse, mixed small and large cell, B-cell type. However, the size of the tumor remained almost stable (16 mm in diameter) over a period of 7 months after diagnosis, without any treatment for lymphoma, indicating a low grade malignancy. We document hepatic lymphoma as an additional complication of primary biliary cirrhosis. PMID- 10364043 TI - Achalasia and Down's syndrome: coincidental association or something else? AB - Achalasia is an uncommon esophageal motor disorder. It has been associated with other diseases such as Parkinson's disease and depressive disorders, but coincidence of achalasia and Down's syndrome is rare. We report five cases of achalasia in Down's syndrome patients seen in our institution. Two of the five cases were diagnosed at pediatric age. Respiratory symptoms and growth retardation were the main clinical manifestations in pediatric patients, whereas adult patients mainly complained of dysphagia. Taking into account the prevalence rate of both disorders, the association seems higher than that expected by chance. The possible etiopathogenic implications of this association, as well as its clinical relevance, are discussed. PMID- 10364044 TI - Gastric carcinoma with osteoclast-like giant cells. AB - Extraskeletal neoplasms with osteoclast-like giant cells are uncommon. These tumors are most frequently reported in the breast and pancreas, and are relatively rare in other sites. We report a case of primary gastric adenocarcinoma with an infiltrate of osteoclast-like giant cells. The patient is a 64-yr-old black woman who presented with epigastric pain and was found to have a mass in the gastric antrum. Histological examination showed a poorly differentiated adenocarcinoma with an infiltrate of osteoclast-like giant cells. The giant cells were present both in the primary gastric adenocarcinoma and in the lymph node metastases. Immunohistochemical stains demonstrated that the giant cells were of monocytic/histiocytic origin and probably represent a distinctive host response to the tumor. The patient is alive and well 12 months after resection. This is the second published report of gastric carcinoma with osteoclast-like giant cells. Based on this limited experience, gastric carcinoma with osteoclast-like giant cells may represent a distinct clinicopathological entity with a more favorable prognosis. PMID- 10364046 TI - Photoallergic skin reaction to ribavirin. AB - A 65-yr-old woman with chronic hepatitis C was treated with three million units interferon-alpha t.i.w. and 1000 mg ribavirin daily. At wk 16 of combination therapy the patient developed an itchy eczematous erythema, partly of urticarial character, which was almost confined to ultraviolet (UV)-exposed sites. Histopathological examination of the skin lesions was consistent with a photoallergic reaction. The minimal erythematous dose for UVA and UVB was assessed on healthy skin. After 24 h, a distinct erythema at the UVB irradiated site was found, whereas no reaction was seen with UVA provocation up to a dose of 10 J/cm2. Correspondingly, determination of the absorption spectrum of ribavirin revealed maximum absorption within UVB at 282.5 nm. Ribavirin was stopped, and the cutaneous lesions and pruritus completely disappeared without subsequent hyperpigmentation. This case indicates that ribavirin is a potential photosensitizer for UVB, which may become increasingly relevant in patients with chronic hepatitis C undergoing combination therapy for 6-12 months with interferon-alpha and ribavirin. PMID- 10364045 TI - Complete remission of multiple hepatocellular carcinomas associated with hepatitis C virus-related, decompensated liver cirrhosis by oral administration of enteric-coated tegafur/uracil. AB - We report a case of complete remission of multiple hepatocellular carcinomas after oral administration of enteric-coated tegafur/uracil. A 77-yr-old woman was diagnosed as having recurrent hepatocellular carcinoma associated with decompensated liver cirrhosis. We administered enteric-coated tegafur/uracil to this patient. After 1 month of oral administration, there was a decrease in tumor markers. An image analysis showed disappearance of hepatocellular carcinoma. No recurrence of the hepatocellular carcinoma was recognized for 18 months up to the time of the patient's death, which was due to massive bleeding from a hemorrhagic rectal ulcer. At autopsy, the tumor lesion had necrotized. Oral administration of enteric-coated granules containing tegafur/uracil may provide an effective treatment for hepatocellular carcinoma. PMID- 10364047 TI - Successful treatment of chylous ascites secondary to Mycobacterium avium complex in a patient with the acquired immune deficiency syndrome. AB - Chylous ascites is a rare form of ascites, the presence of which generally denotes a very poor long term prognosis. We report the case of a patient with acquired immune deficiency syndrome (AIDS) and massive chylous ascites secondary to Mycobacterium avium complex (MAC) infection, identified in the ascitic fluid by a DNA probe assay. With multidrug anti-MAC therapy the ascites resolved completely, and the patient has survived for >21 months. Diagnosis and treatment of MAC-related chylous ascites are reviewed. PMID- 10364048 TI - Recurrent bleeding from a duodenal plasmacytoma treated successfully with embolization of the gastroduodenal artery. PMID- 10364049 TI - Adenocarcinoma of the colon with neuroendocrine features and secretory diarrhea. AB - We report the case of a 63-yr-old man who had severe secretory diarrhea associated with colonic adenocarcinoma, with a prominent signet ring cell component and numerous endocrine cells as demonstrated by positive chromogranin-A staining. Improvement in the secretory diarrhea by the somatostatin analog Sandostatin suggested that the diarrhea was related to a functional neuroendocrine tumor within the colonic tumor, the first case to be reported in the literature. PMID- 10364050 TI - Subclinical syphilitic hepatitis, which was markedly worsened by a Jarisch Herxheimer reaction. AB - Early syphilitic hepatitis is uncommon and tends to be overlooked. However, the diagnosis of this disease is important, because appropriate treatment results in rapid resolution of the hepatitis. We report a case of subclinical early syphilitic hepatitis exaggerated by a Jarisch-Herxheimer reaction. This reaction helped to realize the diagnosis in this case. PMID- 10364051 TI - A pilot study of iron depletion as adjuvant therapy in chronic hepatitis C patients not responding to interferon. AB - The aim of this study was to assess the efficacy of iron depletion obtained by phlebotomy to enhance interferon response in 11 patients who had failed to respond to a standard 3-month interferon treatment. Despite a significant effect on serum aminotransferase levels, there was no effect on viremia, and iron depletion was unable to trigger interferon response. PMID- 10364052 TI - A radiological "stress test" of the esophagus. PMID- 10364053 TI - Better understanding of the pain of pancreatitis. PMID- 10364054 TI - Plastic stents in the esophagus: is homemade really better? PMID- 10364055 TI - Hazards of formulating new theories about gallbladder (GB) function based on ultrasound volume data. PMID- 10364056 TI - Herpes simplex esophagitis. PMID- 10364057 TI - HCV and diabetes mellitus: influence of nosocomial transmission with the use of a finger stick device. PMID- 10364058 TI - Green tea for remission maintenance in Crohn's disease? PMID- 10364059 TI - Baker's yeast in Crohn's disease--can it kill you? PMID- 10364060 TI - Mutation of UGT1A1 gene in a case of Crigler-Najjar syndrome type II. PMID- 10364061 TI - An infestation due to a Taenia saginata with an atypical localization. PMID- 10364062 TI - Treatment of the "ineradicable" Helicobacter pylori infection. PMID- 10364063 TI - Gastrointestinal endoscopy in premenopausal women with iron deficiency anemia: determination of the best diagnostic approach. PMID- 10364064 TI - Prevalence and clinical significance of TT virus coinfection in patients with chronic hepatitis C treated with interferon. PMID- 10364066 TI - Measuring plasma fibrinogen to predict stroke and myocardial infarction: an update. AB - Plasma fibrinogen is a major determinant of platelet aggregation and blood viscosity. The decrease in plasma fibrinogen by bezafibrate is associated with a decrease in the risk of reinfarctions. To strengthen the predictive value of plasma fibrinogen with respect to cardiovascular risk, we performed a meta analysis of studies conducted between 1984 and 1998. Emphasis has been put on the relationship between high levels of plasma fibrinogen and fatal and/or nonfatal cardiovascular events in both the general population and in patients with previous cardiovascular events. Twenty-two studies (13 prospective, 5 cross sectional, and 4 case-control) addressing the association between fibrinogen plasma concentrations and cardiovascular disease were analyzed. The overall estimate of risk of cardiovascular event in subjects with plasma fibrinogen levels in the higher tertile, was twice as high as that of subjects in the lower one (odds ratio, 1.99; 95% confidence interval, 1.85 to 2.13). High plasma fibrinogen levels were associated with an increased risk of cardiovascular disease in healthy as much as in high-risk individuals. A metaregression showed no confounding effects attributable to selected characteristics of retrieved studies. A subgroup analysis (study design, follow up, mean fibrinogen levels, percentage of smokers, and mean age) allowed us to conclude that fibrinogen is an independent risk factor for cardiovascular disease; that it interacts with major determinants of myocardial and cerebrovascular ischemia; and that, in secondary prevention studies, it enhances by 8% the prediction of future events by established risk factors. Thus, fibrinogen measurements should be encouraged to refine the overall risk profiles of individuals and to better tailor preventive interventions. PMID- 10364067 TI - Plasminogen activator inhibitor type 1 in ischemic cardiomyopathy. PMID- 10364068 TI - Ionizing radiation accelerates aortic lesion formation in fat-fed mice via SOD inhibitable processes. AB - Ionizing radiation promotes formation of reactive oxygen species, including the superoxide anion (O2-). To evaluate whether O2- or O2--mediated perturbations may contribute to the known atherogenic effects of radiation, we examined aortic lesion formation in irradiated C57BL/6 mice and evaluated the effects of CuZn superoxide dismutase (CuZn-SOD) overexpression. Ten-week-old mice were exposed to a 2-, 4-, or 8-Gy dose of 250-keV x-rays to the upper thorax and then placed on a high-fat diet for 18 weeks. Based on quantitative lipid staining of serial sections of the proximal aorta, mean lesion area was increased with increasing radiation dose and was 3-fold greater in 8-Gy-irradiated than sham-irradiated mice (7800+/-2140 versus 2635+/-709 micrometer(2), P<0.05). These effects were absolutely dependent on a high-fat diet, which had to be introduced within 1 to 2 weeks of the radiation exposure, suggesting the early involvement of atherogenic lipoproteins that were elevated in response to the diet. The importance of radiation-induced oxidative stress was supported by the observation of a 2-fold lower mean lesion area in irradiated CuZn-SOD transgenic mice than in their irradiated, nontransgenic littermates (3026+/-1590 versus 6102+/-1834 micrometer(2), P<0.05). Lucigenin-enhanced chemiluminescence, used as an index of aortic O2- concentrations, was significantly elevated in the postradiation period, and this response was reduced in CuZn-SOD transgenics. On the basis of these results, we propose that radiation may be a useful tool for initiating oxidative or redox-regulated events that promote atherogenesis and for testing the antiatherogenic properties of antioxidants. PMID- 10364070 TI - Differentiation of human monocytes to monocyte-derived macrophages is associated with increased lipoprotein lipase-induced tumor necrosis factor-alpha expression and production: a process involving cell surface proteoglycans and protein kinase C. AB - The aim of the present study was to (1) evaluate the responsiveness of human mononuclear cells to lipoprotein lipase (LPL), as assessed by tumor necrosis factor-alpha (TNFalpha) production, during the process of differentiation of monocytes to macrophages, and (2) determine the mechanisms by which LPL exerts its effect on these cells. Treatment of human monocytes with purified endotoxin free bovine LPL (1 microgram/mL) resulted in a 161+/-15% increase in TNFalpha production over control values (P<0.01). A further increase in TNFalpha production was observed after treatment of monocyte-derived macrophages (MDMs) with LPL (490+/-81% over control values, P<0.01). Increased TNFalpha mRNA expression and protein kinase C activity were also observed in LPL-treated human monocytes and MDMs. These LPL effects were abrogated by the specific protein kinase C inhibitor calphostin C (1 micromol/L). Although heparinase totally abolished LPL-induced TNFalpha production in human monocytes, this agent did not significantly inhibit LPL effect in human MDMs. In contrast, treatment of MDMs with chondroitinase suppressed LPL-induced TNFalpha production. Taken together, these data suggest that (1) differentiation of human monocytes to MDMs is associated with increased LPL-induced TNFalpha mRNA expression and production, (2) a protein kinase C-dependent pathway is involved in the induction of TNFalpha by LPL in these cells, and (3) LPL effect is mediated by cell surface proteoglycans. As MDMs secrete LPL in the vascular wall, we propose that LPL, by acting as an autocrine activator of MDM function, may contribute to the high level of TNFalpha found in the atheromatous lesion. PMID- 10364069 TI - Smooth muscle-specific SM22 protein is expressed in the adventitial cells of balloon-injured rabbit carotid artery. AB - During the "response-to-injury" process after a mechanical insult to the porcine coronary arteries, the adventitial cells acquire the structural characteristics of myofibroblasts before being incorporated into smooth muscle (SM) layer. We assessed whether the SM-specific SM22 protein can be used as a tracer of adventitial cell-myofibroblast differentiation in the mild balloon injury of rabbit carotid artery. To achieve this goal, we used 2 monoclonal anti-SM22 antibodies (E-11 and 1-B8) and a molecular probe for the SM22alpha mRNA isoform in immunocytochemical and in situ hybridization experiments. The differentiation profile and the migratory and proliferative ability of activated adventitial cells were evaluated by a panel of antibodies to some SM and nonmuscle antigens and pulse- and end-labeling with bromo-deoxyuridine, respectively. In adventitial cells, SM22 antigenicity and SM22alpha mRNA were detectable at days 2 and 4 and, to a lesser extent, at days 7 and 21 after injury, particularly near the adventitia-media interface and mostly colocalizing with bromo-deoxyuridine positive cells. The pulse-labeling experiments showed that the large majority of these cells penetrated the outermost layer of the tunica media without migrating to the subendothelial region. The phenotypic features of activated migrating and nonmigrating adventitial cells resembled those of vimentin-actin myofibroblast subtype and fetal-type SM cells. These findings indicate that a direct exposure of adventitia to the lumen is not required for phenotypic changes and proliferation/migration of these cells. After comparison of the SM22 expression in arterial vessels during early stages of development, we hypothesize that in the injured carotid artery the mural incorporation of adventitial cells and the spatiotemporal activation of SM22 expression are reminiscent of the vascular morphogenetic process and suggest the existence of a stem cell-like reservoir in adventitia. The early adventitial upregulation of SM22 expression in the injured vessel might be related to a multistep transition process in which nonmuscle cells are converted to myofibroblasts and, possibly, to SM cells. PMID- 10364071 TI - Gallates inhibit cytokine-induced nuclear translocation of NF-kappaB and expression of leukocyte adhesion molecules in vascular endothelial cells. AB - Gallates (gallic acid esters) belong to the class of phenolic compounds, which are abundant in red wine. In this study, we show that gallates can inhibit cytokine-induced activation of nuclear factor kappaB (NF-kappaB) and thereby reduce expression of endothelial-leukocyte adhesion molecules in cultured human umbilical vein endothelial cells (HUVECs). Pretreatment of HUVECs with ethyl gallate (3 to 10 micromol/L) significantly suppressed interleukin-1alpha (IL 1alpha)- or tumor necrosis factor-alpha (TNF-alpha)- induced mRNA and cell surface expression of vascular cell adhesion molecule 1 (VCAM-1), intercellular adhesion molecule 1 (ICAM-1), and E-selectin, which was associated with reduced adhesion of leukocytes to HUVECs. Gel shift assays with the NF-kappaB consensus sequence showed the decreased densities of the shifted bands in gallate-treated HUVECs. Furthermore, gallate pretreatment inhibited cytokine-induced transcription of a fusion gene, which consisted of 4 repeats of the NF-kappaB consensus sequence and the luciferase reporter gene. Immunoblot analysis of nuclear extracts and whole-cell lysates demonstrated the decreased amounts of NF kappaB p65 in nuclei but equal amounts of inhibitor-kappaBalpha (I-kappaBalpha) in whole-cell lysates of ethyl gallate-treated HUVECs. Incubation of the nuclear extracts from cytokine-activated HUVECs with ethyl gallate did not affect the NF kappaB shifted bands induced by cytokines in gel shift assays. Taken together, these data demonstrate that ethyl gallate can inhibit cytokine-induced nuclear translocation of NF-kappaB p65 by way of a mechanism independent of I-kappaBalpha degradation and thereby suppress expression of VCAM-1, ICAM-1, and E-selectin, which was associated with reduced adhesion of leukocytes. These results in vitro demonstrate that gallates can exhibit anti-inflammatory properties by blocking activation of NF-kappaB and suggest that these natural compounds, abundant in red wine, may play important roles in the prevention of atherosclerosis and inflammatory responses in vivo. PMID- 10364072 TI - Stat6 activation is essential for interleukin-4 induction of P-selectin transcription in human umbilical vein endothelial cells. AB - Chronic upregulation of P-selectin expression on the surface of the endothelium has been observed in and likely contributes to a number of chronic inflammatory diseases, including atherosclerosis. Agonists of P-selectin expression fall into 2 categories: those that induce a very rapid, transient increase, lasting only hours, and those that induce prolonged upregulation lasting days. It is the latter group, which includes interleukin-4 (IL-4), that is likely to be a mediator of chronic P-selectin upregulation. The increase in P-selectin expression induced by IL-4 results from increased transcriptional activation of the P-selectin gene. The aim of this study was to deduce the postreceptor signaling pathway(s) giving rise to the prolonged increase in P-selectin expression induced by IL-4. We demonstrate the existence of 2 functional signal transducer and activator of transcription 6 (Stat6) binding sites on the P selectin promoter and further demonstrate, by functional analysis of the P selectin promoter, that binding of activated Stat6 to at least 1 site is essential for IL-4-induction of P-selectin transcription. Site 1 (nucleotide[nt] 142) bound Stat6 with a higher affinity than did site 2 (nt -229), and this difference was reflected functionally as constructs in which only site 1 was functional showed full IL-4 inducibility, whereas constructs in which only site 2 was functional showed only 40% of maximal IL-4 inducibility. IL-4 also induced prolonged activation of Stat6, which was contingent on the continuous presence of IL-4. The sustained activation of Stat6 induced by IL-4 is likely to be a key factor leading to the prolonged activation of the P-selectin promoter, thereby resulting in prolonged P-selectin upregulation. PMID- 10364073 TI - Retinoic acid regulates arterial smooth muscle cell proliferation and phenotypic features in vivo and in vitro through an RARalpha-dependent signaling pathway. AB - We have recently shown that all-trans retinoic acid (tRA) modulates arterial smooth muscle cell (SMC) morphologic features and biochemical composition in vitro. It has been proposed that different SMC phenotypes coexist in arteries, which may be retrieved in culture: hence, a differential action of tRA on distinct SMC subsets is conceivable. We have examined the effect of tRA on SMC proliferation, migration, plasminogen activator activity, and alpha-smooth muscle actin expression in 2 phenotypically different rat SMC populations, cultured respectively from the normal aortic media and from the intimal thickening (IT) after endothelial injury. tRA inhibited proliferation and increased migration and tissue-type plasminogen activator activity in both SMC populations, but decreased alpha-smooth muscle actin only in SMC cultured from the IT. The action of tRA is mediated by 2 families of nuclear receptors, RAR and RXR, each containing 3 isoforms, alpha, beta, and gamma. RAR and RAR-alpha agonists, but not RXR agonists, inhibited SMC proliferation in both cell populations and alpha-smooth muscle actin expression only in IT SMC. When administered intraperitoneally to balloon-injured rats, tRA and RAR-alpha agonists reduced the intimal hyperplasia in the carotid artery. Our results show that tRA and synthetic retinoids can affect the proliferation, migration, and differentiation of SMC in vitro. Furthermore, retinoids are able to reduce the IT induced by endothelial injury in vivo. PMID- 10364074 TI - All ApoB-containing lipoproteins induce monocyte chemotaxis and adhesion when minimally modified. Modulation of lipoprotein bioactivity by platelet-activating factor acetylhydrolase. AB - Mildly oxidized LDL has many proinflammatory properties, including the stimulation of monocyte chemotaxis and adhesion, that are important in the development of atherosclerosis. Although ApoB-containing lipoproteins other than LDL may enter the artery wall and undergo oxidation, very little is known regarding their proinflammatory potential. LDL, IDL, VLDL, postprandial remnant particles, and chylomicrons were mildly oxidized by fibroblasts overexpressing 15 lipoxygenase (15-LO) and tested for their ability to stimulate monocyte chemotaxis and adhesion to endothelial cells. When conditioned on 15-LO cells, LDL, IDL, but not VLDL increased monocyte chemotaxis and adhesion approximately 4 fold. Chylomicrons and postprandial remnant particles were also bioactive. Although chylomicrons had a high 18:1/18:2 ratio, similar to that of VLDL, and should presumably be less susceptible to oxidation, they contained (in contrast to VLDL) essentially no platelet-activating factor acetylhydrolase (PAF-AH) activity. Because PAF-AH activity of lipoproteins may be reduced in vivo by oxidation or glycation, LDL, IDL, and VLDL were treated in vitro to reduce PAF-AH activity and then conditioned on 15-lipoxygenase cells. All 3 PAF-AH-depleted lipoproteins, including VLDL, exhibited increased stimulation of monocyte chemotaxis and adhesion. In a similar manner, lipoproteins from Japanese subjects with a deficiency of plasma PAF-AH activity were also markedly more bioactive, and stimulated monocyte adhesion nearly 2-fold compared with lipoproteins from Japanese control subjects with normal plasma PAF-AH. For each lipoprotein, bioactivity resided in the lipid fraction and monocyte adhesion could be blocked by PAF-receptor antagonists. These data suggest that the susceptibility of plasma lipoproteins to develop proinflammatory activity is in part related to their 18:1/18:2 ratio and PAF-AH activity, and that bioactive phospholipids similar to PAF are generated during oxidation of each lipoprotein. Moreover, LDL, IDL, postprandial remnant particles, and chylomicrons and PAF-AH-depleted VLDL all give rise to proinflammatory lipids when mildly oxidized. PMID- 10364075 TI - A novel mutant, ApoA-I nichinan (Glu235-->0), is associated with low HDL cholesterol levels and decreased cholesterol efflux from cells. AB - A novel variant of apolipoprotein (apo) A-I associated with low high density lipoprotein (HDL) cholesterolemia has been identified in a Japanese family during screening for apoA-I variants by isoelectric focusing (IEF) gel analysis. ApoA-I (Glu235-->0) Nichinan was caused by a 3-bp deletion of nucleotides 1998 through 2000 in exon 4 of the apoA-I gene. Four subjects in the family were heterozygous carriers for this mutation; the mean plasma concentrations of apoA-I and HDL cholesterol of affected family members were 30% and 32% lower, respectively, than those of unaffected family members. There were no differences in the levels of very low density lipoprotein and low density lipoprotein cholesterol, triglycerides, and other apolipoproteins between the carriers and the noncarrier family members. In the proband, plasma lecithin:cholesterol acyltransferase activity was normal. Functional consequences of the mutation were examined by expressing the mutated and wild-type proapoA-I cDNAs in Escherichia coli. Cholesterol efflux to recombinant proapoA-I Nichinan from mouse peritoneal macrophages loaded with [3H]cholesterol-labeled acetylated low density lipoprotein was decreased by 54% when compared that of normal recombinant proapoA I. In vivo turnover studies in normal rabbits demonstrated that the recombinant proapoA-I Nichinan was rapidly cleared (22% faster) compared with normal recombinant proapoA-I. We conclude that apoA-I (Glu235-->0) Nichinan induced a critical structural change in the carboxyl-terminal domain of apoA-I for cellular cholesterol efflux and increased the catabolism of apoA-I, resulting in low HDL cholesterol levels. PMID- 10364076 TI - SREBP-1 binds to multiple sites and transactivates the human ApoA-II promoter in vitro : SREBP-1 mutants defective in DNA binding or transcriptional activation repress ApoA-II promoter activity. AB - -Screening of an expression human liver cDNA library resulted in the isolation of several cDNA clones homologous to sterol regulatory element-binding protein-1 (SREBP-1) that recognize the regulatory element AIIAB and AIIK of the human apoA II promoter. DNaseI footprinting of the apoA-II promoter using SREBP-1 (1 to 460) expressed in bacteria identified 5 overall protected regions designated AIIAB ( 64 to -48), AIICD (-178 to -154), AIIDE (-352 to -332), AIIHI (-594 to -574), and AIIK (-760 to -743). These regions contain inverted E-box palindromic or direct repeat motifs and bind SREBP-1 with different affinities. Transient cotransfection experiments in HepG2 cells showed that SREBP-1 transactivated the 911/29 apoA-II promoter 3.5-fold as well as truncated apoA-II promoter segments that contain 1, 2, 3, or 4 SREBP binding sites. Mutagenesis analysis showed that transactivation by SREBP was mainly affected by mutations in element AIIAB. Despite the strong transactivation of the apoA-II promoter by SREBP-1 we could not demonstrate significant changes on the endogenous apoA-II mRNA levels of HepG2 cells after cotransfection with SREBP-1 or in the presence or absence of cholesterol and 25-OH-cholesterol. An SREBP-1 mutant lacking the amino-terminal activation domain bound normally to its cognate sites and repressed the apoA-II promoter activity. Repression was also caused by specific amino acid substitutions of Leu, Val, or Gly for Lys359, which affected DNA binding. Repression by the DNA binding-deficient mutants was abolished by deletion of the amino-terminal activation domain (1 to 90) of SREBP-1. Overall, the findings suggest that the wild-type SREBP-1 can bind and transactivate efficiently the apoA-II promoter in cell culture. SREBP-1 mutants lacking the activation domain bind to their cognate sites and directly repress the apoA-II promoter whereas mutants defective in DNA binding indirectly repress the apoA-II promoter activity, possibly by a squelching mechanism. PMID- 10364077 TI - Dietary beta-carotene and alpha-tocopherol combination does not inhibit atherogenesis in an ApoE-deficient mouse model. AB - Although lipid oxidation plays a major role in atherogenesis, the role of antioxidants in the prevention and treatment of the process is not clear. Apolipoprotein (apo) E-deficient mice develop spontaneous atherosclerotic lesions in major arteries. The presence of oxidized lipoprotein epitopes in the lesion suggests that oxidation reactions are involved in atherogenesis in this mouse model, but the inhibitory effect of antioxidants on atherogenesis in the model is controversial. To test the effect of dietary antioxidants on atherogenesis, male apoE-deficient mice (n=15) were fed a standard chow diet supplemented with 0.05% alpha-tocopherol and 0.05% all-trans beta-carotene. A control group (n=15) received no antioxidant supplement. At the end of the trial, mice consuming vitamins had 5x more plasma vitamin E but undetectable beta-carotene levels. However, liver levels of the beta-carotene metabolite, retinyl palmitate, were higher in antioxidant-treated mice compared with control mice. The antioxidants had no effect on lipoprotein or on plasma anti-oxidatively modified low density lipoproteins (anti-oxLDL) antibody levels. The vitamins had a small but insignificant effect on lipoprotein resistance to ex vivo oxidation, determined by a longer lag period of conjugated diene formation. Atherosclerosis, determined by the lesion size at the aortic sinus, was insignificantly suppressed in antioxidant-treated mice (mean area+/-SE, 20 000+/-7129 versus 13 281+/-5861 micrometer(2); P=0.40). The aortic atherosclerotic lesion area was similar in both experimental groups (2.55+/-0.65% and 2.08+/-0.5% of total aortic area in the control and antioxidant group, respectively; P=0.58). The results of the current study suggest that moderate levels of synthetic antioxidant vitamins have no effect on atherogenesis in apoE-deficient mice. PMID- 10364078 TI - Estrogen-mediated increases in LDL cholesterol and foam cell-containing lesions in human ApoB100xCETP transgenic mice. AB - The murine double transgenic mouse expressing both human apoB100 and cholesteryl ester transfer protein (CETP), has been used as a model to understand the effects mediated by various therapeutic modalities on serum lipoproteins and on atherosclerotic lesion progression. In the present study the effects of estrogen therapy on serum lipoproteins were investigated after mice were placed on an atherosclerotic diet. The daily oral administration of 20 or 100 microg/kg of 17 alpha-ethinyl estradiol resulted in a significant, dose-dependent increase in LDL cholesterol over a 20-week regimen. These differences were apparent by 6 weeks and further increases were observed through the 20-week period. Although CETP did result in a reduction in total HDL, estrogen did not have any impact on the amount of CETP activity associated with the HDL particles. The significant increase in LDL cholesterol was associated with increases in the amount of apoB100 and B48 and apoE-containing particles. Hepatic apoB message levels, however, were not different between the experimental groups. Although the extent of atherosclerotic lesions was modest, <0.5% of the aortic surface area in the vehicle group, the high-dose estrogen group, showed an increase in lesion area consistent with the elevation in LDL cholesterol. These lesions, primarily restricted to the aortic root and aortic semilunar valves, were more intensely stained with Oil Red O in the high-dose estrogen group when compared with the vehicle controls. PMID- 10364079 TI - The natural course of atherosclerosis. Part I: incidence and progression. AB - The natural course of early atherogenesis is not well established. The current prospective survey was designed to monitor 5-year changes in carotid atherosclerosis in a large, stratified random sample of the general population using high-resolution duplex ultrasound (Bruneck Study). Incidence rates of carotid atherosclerosis ranged from near zero to 184 per 1000 person-years. Most atherosclerotic lesions developed at sites with enhanced wall thickness. Incidence of atherosclerosis in premenopausal women was less than half of that observed in men of equal age. The sex difference disappeared within 5 years after menopause and may possibly be attributed to sex variations in body iron stores. Preexisting atherosclerotic lesions may experience 1 of 2 different types of disease progression. 1) The first main type of plaque growth causing nonstenotic or diffuse dilative atherosclerosis was characterized by slow and continuous plaque extension, which usually affected several lesions simultaneously and did not primarily focus on the carotid bifurcation. This step-by-step process relied on a cumulative exposure to well-known risk factors such as hyper-lipidemia. Compensatory enlargement of the artery at the site of active atherosclerosis effectively preserved a (near) normal lumen. 2) The second main type of plaque growth was characterized by occasional prominent increases in lesion size. This process primarily occurred in the internal carotid artery and was mediated by procoagulant risk factors in a way that peak levels were relevant rather than cumulative exposure. As the main underlying pathomechanism, atherothrombosis may be hypothesized. Marked increases in plaque size and insufficient vascular remodeling acted synergistically in producing a significant compromise of the lumen. The current study provides novel insights into the natural course of early carotid atherosclerosis, thereby focusing on disease incidence and various types of spontaneous disease progression. Nonstenotic or diffuse dilating atherosclerosis and focal stenotic disease were found to constitute epidemiologically and etiologically distinct disease entities that develop and proceed independently of each other. PMID- 10364080 TI - The natural course of atherosclerosis. Part II: vascular remodeling. Bruneck Study Group. AB - Arterial remodelling is a potentially important component in atherogenesis aimed at delaying the development of significant lumen compromise. Current knowledge on this phenomenon is mainly restricted to experimental evaluations and a few postmortem studies. We used high-resolution duplex ultrasound to study 5-year changes (1990 to 1995) in vessel geometry in a large random sample of the general population (Bruneck Study). Carotid arteries free of atherosclerosis and wall thickening preserved a normal size to high ages. In contrast, common and internal carotid arteries with elevated intima-media thickness (>/=50th percentile) experienced marked age-dependent dilation that started already in the 5th decade and continuously accelerated thereafter (structural ageing). Vessel diameters were subject to complex regulation involving morphometric characteristics, sex, wall thickness, hypertension, LDL cholesterol levels, and alcohol consumption. Vascular remodelling secondary to incident or slowly progressive (mural) atherosclerosis included local compensation and a generalised dilation response of vascular segments not primarily affected. Adaptive enlargement at the site of active atherogenesis effectively preserved a near-normal lumen in most instances. The current study identified a second main type of plaque growth, characterized by episodic marked increase in lesion volume probably on the basis of plaque thrombosis. In this setting, we did not observe maximum but insufficient compensation but instead usually observed no compensation at all. Failure of vascular remodelling and marked expansion in plaque size acted synergistically in producing significant lumen compromise. The current prospective survey describes fundamental principles and various facets of arterial remodelling and vascular biology in the general population (in vivo). Vessel geometry was subject to marked temporal changes and showed a correspondingly complex (multifactorial) and dynamic regulation. Vascular remodelling emerged as an important compensatory process in human atherogenesis, which crucially contributed to the determination of lumen obstruction. Efficacy and failure of compensation primarily depended on the type and pathomechanisms of underlying atherogenesis and only in second instance on plaque size and location. PMID- 10364081 TI - Atrophic remodeling of the artery-cuffed artery. AB - Increased arterial wall tension stimulates growth and remodeling of arteries, but little is known about the effects of decreased wall tension, despite its developmental and pathological significance. Consequently, we cuffed 1 carotid artery in rabbits with a portion of the contralateral artery to off-load circumferential wall tension. The model produced rapid and extensive atrophy of the cuffed artery that yielded decreases in the DNA content of the cuffed artery (a measure of cell number) from 8.0+/-0.5 microgram/cm of in situ vessel length to 5.6+/-0.5 microgram/cm at 21 days postoperatively. The elastin content of the cuffed artery was also significantly reduced, from 399+/-17 to 283+/-17 microgram/cm, and collagen content was reduced from 468.0+/-59.0 to 154+/-24 microgram/cm (P<0.05) at 21 days postoperatively. Detection of DNA oligonucleosomes by gel electrophoresis implicated apoptotic cell death in remodeling due to cuffing. Upregulation of matrix metalloproteinases (MMPs), including MMP-2, MMP-9, and unidentified gelatinases, indicated that these enzymes may also be involved in remodeling. No further changes in wall structure were seen between 3 weeks and 6 months, and the excised artery that was used as a cuff exhibited normal medial morphology for at least 6 months postoperatively. We infer from these experiments that off-loading of arterial wall tension induces rapid and extensive atrophy of the arterial media. PMID- 10364082 TI - Complete processing of type III collagen in atherosclerotic plaques. AB - The extent of processing of type III collagen is assessed, and the proportions of type I and III collagens are estimated in atherosclerotic plaques obtained from the carotid artery, common femoral artery, and aorta. The fraction of type III collagen that had retained its amino-terminal propeptide (pN-collagen) was 42% in the soluble extract but only 0.0081% in the insoluble residue. Taken together, only 0.011% of the type III collagen in whole plaques was in the form of type III pN-collagen. Together with the small amounts of the free propeptides of type I procollagen, this finding indicates a low rate of collagen turnover. The amounts of solubilized telopeptides of type I and III collagens were measured, after heat denaturation and trypsin digestion of the collagenous helix, by specific immunoassays for the corresponding trypsin-generated antigens. The mean proportion of type III collagen was 61% (95% confidence interval, 58% to 65%) in the carotid and femoral artery plaques and 56% (95% confidence interval, 44% to 68%) in the aortic specimens. The completely processed and cross-linked type III collagen seems to be the major collagen type in atherosclerotic plaques. PMID- 10364083 TI - Relationship between lipoprotein- and oxidation-related variables and atheroma lipid composition in subjects undergoing coronary artery bypass graft surgery. AB - The relationship between atheroma lipid composition and serum lipoprotein and oxidation measurements has not been fully explored. To address this question, we studied serum, plasma, and aortic wall specimens from 66 subjects undergoing coronary artery bypass graft surgery. The lipid composition of aortic specimens was characterized in terms of cholesterol ester and cholesterol crystal plus phospholipid by using hot-stage polarizing light microscopy; tissue oxidation status was assessed by measuring conjugated dienes. Serum lipoprotein-related measurements included total cholesterol, triglyceride, low density lipoprotein (LDL) cholesterol, high density lipoprotein (HDL) cholesterol, apolipoproteins B and AI, and lipoprotein(a). Oxidation status was assessed by measuring LDL mobility, thiobarbituric acid-reactive substances, LDL conjugated dienes, and IgG and IgM autoantibodies against oxidized LDL. Fasting blood glucose was also determined. Lesion cholesterol crystal plus phospholipid content was associated inversely with serum HDL cholesterol levels (r=-0.279, P=0.029) and positively with fasting blood glucose (r=0.359, P=0.016), LDL mobility (0.276, P<0.05), and IgM autoantibodies against oxidized LDL (r=0.272, P=0.037). There was also a significant relationship between the level of aortic tissue conjugated dienes and plasma LDL mobility (r=0.332, P=0.007). In multivariate analysis, IgM autoantibodies against oxidized LDL, fasting blood glucose, and LDL mobility, in descending order of significance, together accounted for 35% of the variability in aortic lesion cholesterol crystal plus phospholipid content. These data support direct and independent roles for oxidation and hyperglycemia in the pathophysiology of atherosclerosis. PMID- 10364084 TI - Monocyte chemoattractant protein-1 accelerates atherosclerosis in apolipoprotein E-deficient mice. AB - The pro-inflammatory chemokine, monocyte chemoattractant protein-1 (MCP-1), plays a fundamental role in monocyte recruitment and has been implicated as a contributing factor to atherosclerosis. The predominant cell types within the vessel wall--endothelial cells, smooth muscle cells, and macrophages--all contribute to overexpression of MCP-1 in atherosclerotic tissue. In this report we assess the role of MCP-1 expression by leukocytes on lesion progression in a murine model susceptible to atherosclerosis. Bone marrow cells from mice overexpressing a murine MCP-1 transgene on a background of apoE-deficiency or from control mice were transplanted into irradiated apoE-knockout mice. After repopulation of apoE-knockout mice with bone marrow containing the MCP-1 transgene, macrophages expressing the MCP-1 transgene were found in several tissues, including the aorta. Qualitative assessment of atherosclerosis in these mice revealed increased lipid staining, a 3-fold (P<0.001) increase in the amount of oxidized lipid, and increased immunostaining for macrophage cell surface markers with anti-F4/80 and anti-CD11b antibodies. There were no differences in plasma lipids, plasma lipoprotein profiles, or body weight between the 2 groups. These results provide the first direct evidence that MCP-1 expression by leukocytes, predominately macrophages, increases the progression of atherosclerosis by increasing both macrophage numbers and oxidized lipid accumulation. PMID- 10364085 TI - Localization of lipoprotein lipase in the diabetic heart: regulation by acute changes in insulin. AB - Vascular endothelium-bound lipoprotein lipase (LPL) is rate limiting for free fatty acid (FFA) transport into tissues. In streptozotocin (STZ)-diabetic rats, we have previously demonstrated an increased heparin-releasable LPL activity from perfused hearts. Because heparin can traverse the endothelial barrier, conventional Langendorff retrograde perfusion of the heart with heparin could release LPL from both the capillary luminal and abluminal surfaces. To determine the precise location of the augmented LPL, a modified Langendorff retrograde perfusion was used to isolate the enzyme at the coronary lumen from that in the interstitial effluent. In response to heparin, a 4-fold increase in LPL activity and protein mass was observed in the coronary perfusate after 2 weeks of STZ diabetes. Release of LPL activity into the interstitial fluid of control hearts was slow but progressive, whereas in diabetic hearts, peak enzyme activity was observed within 1 to 2 minutes after heparin, followed by a gradual decline. Immunohistochemical studies of myocardial sections confirmed that the augmented LPL in diabetic hearts was mainly localized at the capillary endothelium. To study the acute effects of insulin on endothelial LPL activity, we examined rat hearts at various times after the onset of hyperglycemia. An increased heparin releasable LPL activity in diabetic rats was demonstrated shortly (6 to 24 hours) after STZ injection or after withdrawal from exogenous insulin. Heparin releasable coronary LPL activity was also increased after an overnight fast. These studies indicate that the intravascular heparin-releasable fraction of cardiac LPL activity is acutely regulated by short-term changes in insulin rather than glucose. Thus, during short periods (hours) of hypoinsulinemia, increased LPL activity at the capillary endothelium can increase the delivery of FFAs to the heart. The resultant metabolic changes could induce the subsequent cardiomyopathy that is observed in the chronic diabetic rat. PMID- 10364086 TI - Mutations in the lipoprotein lipase gene associated with ischemic heart disease in men. The Copenhagen city heart study. AB - The aim of this study was to test the hypothesis that the Asp9Asn substitution and the T(-93)-->G mutation in the promoter of the lipoprotein lipase gene affect plasma lipid levels and thereby the risk of ischemic heart disease (IHD). We genotyped 9033 men and women from a general population sample and 940 patients with IHD. The frequency of both the G allele and the Asn9 allele in the general population sample was approximately 0.015 for both men and women. These 2 mutations appeared together in 95% of carriers. The average triglyceride-raising effect associated with double heterozygosity for the T(-93)-->G mutation and the Asp9Asn substitution was 0.28 mmol/L (P=0.004) and 0.16 mmol/L (P=0.10) in men and women, respectively. On logistic regression analysis allowing for age, the risk of IHD for double heterozygous men and women was increased 90% (95% confidence interval [CI], 20% to 200%) and 30% (95% CI, -40% to 170%), respectively, compared with noncarriers. When, in addition, other conventional cardiovascular risk factors were allowed for, the risk of IHD for double heterozygous men and women was increased 70% (95% CI, 0% to 190%) and 20% (95% CI, -50% to 180%), respectively. Of the overall risk of IHD in men in the general population, the fraction attributable to double heterozygosity was 3%, similar to the 5% attributable to diabetes mellitus. These results demonstrate that the Asp9Asn substitution is in linkage disequilibrium with the T(-93)-->G mutation and that the double-heterozygous carrier status is associated with elevated plasma triglycerides and an increased risk of IHD in men. PMID- 10364087 TI - Effects of lovastatin therapy on susceptibility of LDL to oxidation during alpha tocopherol supplementation. AB - A randomized, double-masked, crossover clinical trial was carried out to evaluate whether lovastatin therapy (60 mg daily) affects the initiation of oxidation of low density lipoprotein (LDL) in cardiac patients on alpha-tocopherol supplementation therapy (450 IU daily). Twenty-eight men with verified coronary heart disease and hypercholesterolemia received alpha-tocopherol with lovastatin or with dummy tablets in random order. The two 6-week, active-treatment periods were preceded by a washout period of at least 8 weeks. The oxidizability of LDL was determined by 2 methods ex vivo. The depletion times for LDL ubiquinol and LDL alpha-tocopherol were determined in timed samples taken during oxidation induced by 2, 2-azobis(2,4-dimethylvaleronitrile). Copper-mediated oxidation of LDL isolated by rapid density-gradient ultracentrifugation was used to measure the lag time to the propagation phase of conjugated-diene formation. alpha Tocopherol supplementation led to a 1.9-fold concentration of reduced alpha tocopherol in LDL (P<0.0001) and to a 2.0-fold longer depletion time (P<0.0001) of alpha-tocopherol compared with determinations after the washout period. A 43% prolongation (P<0.0001) was seen in the lag time of conjugated-diene formation. Lovastatin decreased the depletion time of reduced alpha-tocopherol in metal ion independent oxidation by 44% and shortened the lag time of conjugated-diene formation in metal ion-dependent oxidation by 7%. In conclusion, alpha-tocopherol supplementation significantly increased the antioxidative capacity of LDL when measured ex vivo, which was partially abolished by concomitant lovastatin therapy. PMID- 10364088 TI - Minimal platelet deposition and activation in models of injured vessel wall ensure optimal neutrophil adhesion under flow conditions. AB - Platelets at injured vessel wall form an adhesive surface for leukocyte adhesion. The precise relation between platelet adhesion and activation and leukocyte adhesion, however, is not known. We therefore used various models of injured vessel wall to form different patterns of platelet adhesion. The interaction of polymorphonuclear neutrophils (PMNs) was subsequently studied under flow conditions. In the absence of platelets, not only endothelial cell, smooth muscle cell, and fibroblast matrices but also purified matrix proteins (fibrinogen, collagen, and fibronectin) barely support PMN adhesion. The presence of platelets, however, strongly enhances PMN adhesion. PMN adhesion shows a proportional increase with platelet coverage up to 15%. Although PMNs roll over the scarcely scattered platelets, they speed up again when encountering surfaces without platelets. This "hopping" interaction of PMNs vanishes with platelet coverage >15%. Unobstructed rolling of PMNs is than observed and soon leads to a maximal adhesion of 1000 to 1200 cells/mm2. The mean rolling velocity of PMNs continues to decrease with higher platelet coverage. Platelet aggregate formation is an accepted indicator of platelet activation. The presence of platelet aggregates instead of contact or spread platelets, however, does not increase PMN adhesion. Also, additional stimulation of surface-associated platelets by thrombin fails to influence PMN adhesion. Moreover, indomethacin as an inhibitor of platelet activation and aggregation does not change the subsequent PMN interaction. In conclusion, approximately 15% of platelet coverage is sufficient for optimal PMN adhesion. Increasing platelet coverage increases the availability of platelet-associated receptors that lower PMN rolling velocity. Additional activation of adherent platelets makes no difference in the expression of relevant adhesion receptors. Therefore, minimal vascular damage in vivo and only scarce platelet adhesion will already evoke significant colocalization of leukocytes. PMID- 10364089 TI - Beneficial effects of conversion from cyclosporine to azathioprine on fibrinolysis in renal transplant recipients. AB - Cyclosporin A (CsA) has been implicated as one of the factors contributing to the high cardiovascular morbidity and mortality after renal transplantation. This may be mediated by either a high prevalence of conventional risk factors for atherosclerosis, such as hypertension, hypercholesterolemia, and diabetes mellitus, or by impairment of the fibrinolytic activity evoked by CsA, possibly through interference with prostanoid metabolism. We therefore assessed the impact of conversion of CsA to azathioprine immunosuppressive treatment on parameters of fibrinolytic activity and plasma concentration of the prostanoids prostaglandin E2 and thromboxane B2 in 18 stable renal transplant recipients. During CsA, mean arterial pressure and serum creatinine were significantly higher than during azathioprine (116+/-15 mm Hg versus 106+/-13 mm Hg, P=0.0003; and 147+/-34 micromol/L versus 127+/-35 micromol/L, P=0.002; mean+/-SD). On conversion, the plasma tissue plasminogen activator activity increased from 1.2 (1.1 to 1.7; median, 95% CI) to 1.8 (1.6 to 2.0) IU/mL (P=0.011), without a significant change of the plasminogen activator antigen concentration. This was associated with a substantial decrease in plasminogen activator inhibitor-1 activity from 10.4 (8.5 to 16.7) to 6.4 (5.6 to 9.2) IU/mL (P=0.009). Furthermore, plasma levels of prostaglandin E2 and thromboxane B2 markedly decreased (from 9.7 [7.4 to 12.9] to 4.6 [4.3 to 8.1] pg/mL, P=0.0006; and from 106.1 [91.7 to 214.2] to 70.2 [50.3 to 85.6] pg/mL, P=0.002, respectively). During CsA, but not azathioprine, plasma tissue plasminogen activator antigen and plasminogen activator inhibitor-1 levels correlated significantly with prostaglandin E2 (r=0.53, P=0.02; and r=0.60, P=0.008, respectively), and thromboxane B2 (r=0.75, P=0.0001; and r=0.77, P=0.0001, respectively) levels. In conclusion, CsA induced substantial impairment of fibrinolytic activity, which recovered after conversion to azathioprine. The impaired fibrinolysis observed during CsA treatment may be caused by modulation of eicosanoid production or metabolism in vascular endothelial cells and possibly contributes to the high incidence of cardiovascular disease after kidney transplantation. PMID- 10364090 TI - Effect of strenuous, acute exercise on alpha2-adrenergic agonist-potentiated platelet activation. AB - Vigorous exercise transiently increases the risk of primary cardiac arrest. Strenuous, acute exercise can also increase the release of plasma epinephrine. Previous investigations have indicated that epinephrine can potentiate platelet activation by activating platelet alpha2-adrenoceptors. This study investigated how strenuous, acute exercise affects alpha2-adrenergic agonist-potentiated platelet activation by closely examining 15 sedentary men who exercised strenuously on a bicycle ergometer. Before and immediately after exercise, platelet adhesiveness on fibrinogen-coated surfaces, [Ca2+]i in platelets, the number and affinity of alpha2-adrenergic sites on the platelet surface, and plasma catecholamine levels were determined. The results of this study can be summarized as follows: (1) The affinity of alpha2-adrenergic receptors on platelets decreases while the maximal binding number significantly increases after strenuous exercise, thereby correlating with the rise in plasma catecholamine levels. (2) Basal, clonidine-treated, ADP-treated, and clonidine plus ADP-treated adhesiveness and [Ca2+]i in platelets increased after strenuous exercise. (3) Strenuous exercise is associated with higher percentages of ADP- and clonidine plus ADP-enhanced platelet adhesiveness and [Ca2+]i than at rest. (4) The synergistic effects of clonidine on ADP-enhanced platelet adhesiveness and [Ca2+]i after strenuous exercise are much greater than those at rest. Therefore, we conclude that strenuous, acute exercise enhances platelet activation, possibly by altering the performance of platelet alpha2-adrenergic receptors, facilitating the ability of ADP-activated fibrinogen receptors, and enhancing fibrinogen binding to platelet fibrinogen receptors. PMID- 10364091 TI - Influence of antithrombin III on coagulation and inflammation in porcine septic shock. AB - The physiological inhibitor of thrombin, antithrombin III (ATIII, Kybernin P) was investigated for its antiinflammatory and anticoagulant effects in a pig model of septic shock. Pigs were infused with a dose of 0.25 microgram. kg-1. h-1 of lipopolysaccharide (LPS) over a period of 3 hours. Animals developed systemic inflammation, disseminated intravascular coagulation (DIC), organ failure and cardiovascular abnormalities, namely pulmonary hypertension and systemic hypotension. Twenty septic pigs were allocated to 2 study groups, treated either with ATIII (n=10) or placebo (n=10). ATIII was administered as a 250-U/kg IV bolus infusion for 30 minutes (-60 to -30 minutes) followed by a single IV bolus of 125 U/kg (t=0) and a second 30-minute infusion of 250 U/kg (120 to 150 minutes). ATIII significantly prevented the development of a DIC; the increase in fibrin monomers (placebo, 11.4+/-9.1 reciprocal titers, at 6 hours) was completely overcome by ATIII (P<0. 05). ATIII significantly prevented the increase in thromboxane (TXB2) levels, which were 809+/-287 pg/mL in the placebo and 420+/-174 pg/mL in the verum group after 6 hours (P<0.02). On the other hand, ATIII had no influence on TNF levels. In a lethal study with an increased dose of LPS (0.5 microgram. kg-1. h-1). A significant reduction in mortality was observed in the ATIII group (0 of 7) compared with the placebo group (4 of 6) (P<0.05, chi2 test) a significant reduction of pulmonary hypertension (placebo, 42.0+/-11. 1 mm Hg; ATIII, 23.6+/-7.5 mm Hg, P<0.05), but no effect on systemic hypotension, was noted in the ATIII group. It was thus concluded that modulation of the procoagulatory state by substitution of ATIII results in a late beneficial antiinflammatory effect in this model of septic shock. PMID- 10364092 TI - Complex association of protein C gene promoter polymorphism with circulating protein C levels and thrombotic risk. AB - The allele and haplotype frequency of the -1654 C/T and -1641 A/G protein C (PC) gene promoter polymorphisms was determined and analyzed according to circulating PC concentrations in 394 healthy subjects aged 20 to 60 years. The CG haplotype was associated with a lower PC concentration in both homozygous and heterozygous subjects compared with noncarriers. The TA allele had the reverse effect, but only in homozygotes. The distribution of the CG and TA alleles was significantly different in 242 patients, aged 17 to 60 years, with venous thromboembolism. The CG allele increased the risk of thrombosis, with an OR of 1.39 (95% confidence interval (CI), 1.04 to 1.87). The TA allele was protective in subjects aged <45 years, with an OR of 0.68 (95% CI, 0.44 to 1.04). TA was also significantly associated with older age at the first thrombosis. This study confirms the link between the PC gene promoter and circulating PC levels, and suggests a complex effect on the risk of thrombosis. PMID- 10364093 TI - Effects of fibrate compounds on expression of plasminogen activator inhibitor-1 by cultured endothelial cells. AB - The consistent positive correlation between triglyceride and plasminogen activator inhibitor-1 (PAI-1) levels in plasma and the fact that very low density lipoprotein (VLDL) induces secretion of PAI-1 from cultured human umbilical vein endothelial cells (HUVECs) and human hepatoblastoma cells have raised the question of whether fibrate treatment, the main effect of which is a profound lowering of plasma concentrations of VLDL, might improve fibrinolytic function by reducing the plasma levels of PAI-1. However, the findings of controlled clinical trials using various fibrate compounds have been discrepant. ECs express PAI-1 under normal conditions in humans. We therefore examined the effects of several fibrate compounds on PAI-1 expression and secretion by cultured HUVECs and the HUVEC-derived cell line EA.hy926. All fibrate compounds examined had significant effects on PAI-1 gene transcription in the EA.hy926 cells. Low concentrations of clofibric acid and bezafibrate increased PAI-1 transcription and secretion, whereas Wy-14643 increased PAI-1 synthesis in a dose-dependent way. In contrast, both fenofibric acid and gemfibrozil markedly decreased PAI-1 transcription and secretion from HUVECs and EA.hy926 cells. Thus, stimulation of the transcriptional activity of the PAI-1 gene by some fibrates is linked to increased secretion of PAI-1 protein by the cells, whereas the opposite effects occur with other fibrate compounds. Whether the different effects on PAI-1 transcription and secretion by ECs in vitro also reflect differences in treatment effects on the regulation of plasma PAI-1 activity in vivo will have to be determined in larger-scale, controlled clinical trials. PMID- 10364094 TI - Impact of adipose tissue on plasma plasminogen activator inhibitor-1 in dieting obese women. AB - The increased incidence of cardiovascular diseases in obese subjects could be partially attributed to impaired fibrinolysis due to elevated plasma levels of tissue plasminogen activator inhibitor 1 (PAI-1). The associations between changes in plasma PAI-1, metabolic variables, and adipose tissue during weight loss and regain were studied in 52 healthy, premenopausal, obese women participating in a weight reduction program with a hypocaloric diet. PAI-1, insulin, triglyceride, leptin, and adipsin levels were determined at entry, after the first week, after completion of the program, and after 5 months of follow-up. In the 33 obese women who completed the program, decreases in PAI-1 antigen ( 54%), PAI activity (-74%), and leptin (-51%), but not of adipsin, were observed. Changes in PAI-1 were associated with changes in body mass index (BMI), body fat, leptin, and insulin. The decreased level of PAI-1 remained low after follow-up in the 14 women who maintained their reduced weight but increased in the 16 women who regained weight. This increase in PAI-1 was correlated with an increase in body fat and leptin. On multivariate analysis, BMI was the major determinant of PAI-1 level. In conclusion, during weight reduction with a hypocaloric diet, the decrease in PAI-1 is more closely related to changes in adipose tissue than to changes in metabolic variables, suggesting a significant role for adipose tissue in regulating plasma levels of PAI-1. PMID- 10364095 TI - Learning from complaints about general practitioners. PMID- 10364096 TI - Free the slaves. PMID- 10364097 TI - Nutritional hyperhomocysteinaemia. PMID- 10364098 TI - Clinical evidence. PMID- 10364100 TI - UK junior doctors vote to ballot on industrial action. PMID- 10364099 TI - Antithrombotic therapy in cancer. PMID- 10364101 TI - Increasing numbers of english junior doctors work above agreed limits on hours PMID- 10364102 TI - In brief PMID- 10364103 TI - European plan to put obesity on governments' health agenda. PMID- 10364104 TI - US considers ban on British blood. PMID- 10364105 TI - Younger women might benefit from breast cancer screening PMID- 10364106 TI - Neural stem cells successfully transplanted PMID- 10364107 TI - Bristol manager argues patient care was a matter for clinicians PMID- 10364108 TI - Welsh office to review consultant's lengthy suspension PMID- 10364109 TI - UK consultants warn of overwork and general demoralisation PMID- 10364110 TI - New president of the german medical council may back reform PMID- 10364112 TI - Doctors in israel oppose open performance data PMID- 10364111 TI - English surgeons call for improved management of head injury. PMID- 10364113 TI - Pregnancy does not increase mortality from breast cancer PMID- 10364114 TI - Another media scare about MMR vaccine hits Britain. PMID- 10364115 TI - Oral contraceptives and myocardial infarction: results of the MICA case-control study. AB - OBJECTIVES: To determine the association between myocardial infarction and use of different types of oral contraception in young women. DESIGN: Community based case-control study. Data from interviews and general practice records. SETTING: England, Scotland, and Wales. PARTICIPANTS: Cases (n=448) were recruited from women aged between 16 and 44 who had suffered an incident myocardial infarction between 1 October 1993 and 16 October 1995. Controls (n=1728) were women without a diagnosis of myocardial infarction matched for age and general practice. MAIN OUTCOME MEASURES: Odds ratios for myocardial infarction in current users of all combined oral contraceptives stratified by their progestagen content compared with non-users; current users of third generation versus second generation oral contraceptives. RESULTS: The adjusted odds ratio for myocardial infarction was 1.40 (95% confidence interval 0.78 to 2. 52) for all combined oral contraceptive users, 1.10 (0.52 to 2.30) for second generation users, and 1.96 (0.87 to 4.39) for third generation users. Subgroup analysis by progestagen content did not show any significant difference from 1, and there was no effect of duration of use. The adjusted odds ratio for third generation users versus second generation users was 1.78 (0.66 to 4.83). 87% of cases were not exposed to an oral contraceptive, and 88% had clinical cardiovascular risk factors or were smokers, or both. Smoking was strongly associated with myocardial infarction: adjusted odds ratio 12.5 (7.29 to 21.5) for smoking 20 or more cigarettes a day. CONCLUSIONS: There was no significant association between the use of oral contraceptives and myocardial infarction. The modest and non-significant point estimates for this association have wide confidence intervals. There was no significant difference between second and third generation products. PMID- 10364116 TI - Experiences of hospital care and treatment seeking for pain from sickle cell disease: qualitative study. AB - OBJECTIVE: To investigate how sociocultural factors influence management of pain from sickle cell disease by comparing the experiences of those who usually manage their pain at home with those who are more frequently admitted to hospital for management of their pain. DESIGN: Qualitative analysis of semistructured individual interviews and focus group discussions. PARTICIPANTS: 57 participants with genotype SS or S/beta-thal (44 subjects) or SC (9) (4 were unknown). 40 participants took part in focus groups, six took part in both focus groups and interviews, and nine were interviewed only. Participants were allocated to focus groups according to number of hospital admissions for painful crisis management during the previous year, ethnic origin, and sex. RESULTS: The relation between patients with sickle cell disease and hospital services is one of several major non-clinical dimensions shaping experiences of pain management and behaviour for seeking health care. Experiences of hospital care show a range of interrelated themes, which are common to most participants across variables of sex, ethnicity, and hospital attended: mistrust of patients with sickle cell disease; stigmatisation; excessive control (including both over- and undertreatment of pain); and neglect. Individuals respond to the challenge of negotiating care with various strategies. Patients with sickle cell disease who are frequently admitted to hospital may try to develop long term relationships with their carers, may become passive or aggressive in their interactions with health professionals, or may regularly attend different hospitals. Those individuals who usually manage their pain at home express a strong sense of self responsibility for their management of pain and advocate self education, assertiveness, and resistance as strategies towards hospital services. CONCLUSIONS: The current organisation and delivery of management of pain for sickle cell crisis discourage self reliance and encourage hospital dependence. Models of care should recognise the chronic nature of sickle cell disorders and prioritise patients' involvement in their care. PMID- 10364119 TI - A new day in surgery PMID- 10364117 TI - Risk factors for erysipelas of the leg (cellulitis): case-control study. AB - OBJECTIVE: To assess risk factors for erysipelas of the leg (cellulitis). DESIGN: Case-control study. SETTING: 7 hospital centres in France. SUBJECTS: 167 patients admitted to hospital for erysipelas of the leg and 294 controls. RESULTS: In multivariate analysis, a disruption of the cutaneous barrier (leg ulcer, wound, fissurated toe-web intertrigo, pressure ulcer, or leg dermatosis) (odds ratio 23.8, 95% confidence interval 10.7 to 52.5), lymphoedema (71.2, 5.6 to 908), venous insufficiency (2.9, 1.0 to 8.7), leg oedema (2.5, 1.2 to 5.1) and being overweight (2.0, 1.1 to 3.7) were independently associated with erysipelas of the leg. No association was observed with diabetes, alcohol, or smoking. Population attributable risk for toe-web intertrigo was 61%. CONCLUSION: This first case control study highlights the major role of local risk factors (mainly lymphoedema and site of entry) in erysipelas of the leg. From a public health perspective, detecting and treating toe-web intertrigo should be evaluated in the secondary prevention of erysipelas of the leg. PMID- 10364118 TI - Preventing injuries in public playgrounds through partnership between health services and local authority: community intervention study. PMID- 10364120 TI - Women's complaints PMID- 10364121 TI - General practitioners' experiences of patients' complaints: qualitative study. AB - OBJECTIVE: To examine how general practitioners experience patients' complaints. SETTING: General practices in Lambeth, Southwark, and Lewisham health authority. PARTICIPANTS: Representative sample of 30 general practitioners who had had complaints made against them under either the old or new complaints system. DESIGN: Qualitative study with detailed interviews. RESULTS: Participants described their experiences of patients' complaints in three stages: initial impact, conflict, and resolution. The first stage described being out of control, feelings of shock and panic, and a sense of indignation towards patients generally. The second stage described the many conflicts generated by the complaint: emotional conflicts such as feelings of anger, depression, and even suicide, conflicts around aspects of professional identity including doubts about clinical competence, conflicts with family and colleagues, and conflicts arising from the management of the complaint. The third stage described a sense of resolution. For many this meant practising defensively, for others it meant planning to leave general practice, and for a minority no resolution was achieved. Not all participants, however, reported such a negative experience. Some described how they had become immune to complaints, and a small minority described the complaint as a learning experience. CONCLUSION: The initial impact stage and conflict stage may be necessary aspects of the experience that general practitioners endure when they have a complaint made against them. Support structures should, however, be in place to help general practitioners through these stages. PMID- 10364123 TI - Lesson of the week: cavernous haemangioma mimicking multiple sclerosis. PMID- 10364124 TI - Email submissions from outside the united kingdom PMID- 10364122 TI - Clinical evidence: atopic eczema. PMID- 10364125 TI - ABC of intensive care: organ dysfunction. PMID- 10364126 TI - Something to celebrate PMID- 10364127 TI - Towards an understanding of oedema. PMID- 10364129 TI - "It's got to be terrible!" PMID- 10364128 TI - How do you choose antibiotic treatment? PMID- 10364130 TI - Is there a rationale for rationing chronic dialysis? Two points need clarification. PMID- 10364131 TI - Rationing of sildenafil. Nobody needs an erection at public expense. PMID- 10364132 TI - Patient centred care of diabetes in general practice. Doctors and nurses must understand meaning of "communication". PMID- 10364133 TI - Prenatal and perinatal risk factors for psychiatric diseases of early onset. Results are different if seasons are categorised differently. PMID- 10364135 TI - Breast feeding and working mothers. Summary of electronic responses. PMID- 10364134 TI - Antibiotics for acute sinusitis in general practice. Entry criteria were too dissimilar for studies to be combined for meta-analysis. PMID- 10364136 TI - Britain's Academy of Medical Sciences has been busy in recent months. PMID- 10364137 TI - Early diet in preterm babies and later intelligence quotient. Surely study showed that breast milk is feed of choice for premature babies. PMID- 10364139 TI - Senior medical staffs conference PMID- 10364138 TI - Robert hugh cawley PMID- 10364140 TI - High life: A history of high altitude physiology and medicine PMID- 10364141 TI - Against the grain: the genetic transformation of global agriculture PMID- 10364142 TI - Adding insult to injury PMID- 10364143 TI - Website of the week: http://www.coventrydoctors.org. uk/westminster/Feedback.htm PMID- 10364144 TI - The beginning of the end PMID- 10364145 TI - Good doctors doing nothing PMID- 10364147 TI - No link supported between myocardial infarction and oral contraceptive use PMID- 10364146 TI - Failure: the great teacher PMID- 10364148 TI - Hospital experiences affect pain management in sickle cell disease PMID- 10364149 TI - Lymphoedema and site of entry are risk factors for cellulitis PMID- 10364151 TI - Patient complaints are damaging for doctors' health and practice PMID- 10364150 TI - Surveillance of playground accidents can lead to their reduction PMID- 10364152 TI - Is choosing the right antibiotic really that difficult? PMID- 10364153 TI - p53-mediated apoptosis is attenuated in Werner syndrome cells. AB - The WRN DNA helicase is a member of the DExH-containing DNA helicase superfamily that includes XPB, XPD, and BLM. Mutations in WRN are found in patients with the premature aging and cancer susceptibility syndrome known as Werner syndrome (WS). p53 binds to the WRN protein in vivo and in vitro through its carboxyl terminus. WS fibroblasts have an attenuated p53- mediated apoptotic response, and this deficiency can be rescued by expression of wild-type WRN. These data support the hypothesis that p53 can induce apoptosis through the modulation of specific DExH containing DNA helicases and may have implications for the cancer predisposition observed in WS patients. PMID- 10364154 TI - FGF signaling inhibits chondrocyte proliferation and regulates bone development through the STAT-1 pathway. AB - Several genetic forms of human dwarfism have been linked to activating mutations in FGF receptor 3, indicating that FGF signaling has a critical role in chondrocyte maturation and skeletal development. However, the mechanisms through which FGFs affect chondrocyte proliferation and differentiation remain poorly understood. We show here that activation of FGF signaling inhibits chondrocyte proliferation both in a rat chondrosarcoma (RCS) cell line and in primary murine chondrocytes. FGF treatment of RCS cells induces phosphorylation of STAT-1, its translocation to the nucleus, and an increase in the expression of the cell-cycle inhibitor p21WAF1/CIP1. We have used primary chondrocytes from STAT-1 knock-out mice to provide genetic evidence that STAT-1 function is required for the FGF mediated growth inhibition. Furthermore, FGF treatment of metatarsal rudiments from wild-type and STAT-1(-/-) murine embryos produces a drastic impairment of chondrocyte proliferation and bone development in wild-type, but not in STAT-1(-/ ) rudiments. We propose that STAT-1 mediated down regulation of chondrocyte proliferation by FGF signaling is an homeostatic mechanism which ensures harmonious bone development and morphogenesis. PMID- 10364155 TI - c-Myc-induced sensitization to apoptosis is mediated through cytochrome c release. AB - Expression of c-Myc sensitizes cells to a wide range of pro-apoptotic stimuli. We here show that this pro-apoptotic effect is mediated through release of mitochondrial holocytochrome c into the cytosol. First, activation of c-Myc triggers release of cytochrome c from mitochondria. This release is caspase independent and blocked by the survival factor IGF-1. Second, c-Myc-induced apoptosis is blocked by microinjection of anticytochrome c antibody. In addition, we show that microinjection of holocytochrome c mimics the effect of c-Myc activation, sensitizing cells to DNA damage and to the CD95 pathway. Both p53 and CD95/Fas signaling have been implicated in c-Myc-induced apoptosis but neither was required for c-Myc-induced cytochrome c release. Nonetheless, inhibition of CD95 signaling in fibroblasts did prevent c-Myc-induced apoptosis, apparently by obstructing the ability of cytosolic cytochrome c to activate caspases. We conclude that c-Myc promotes apoptosis by causing the release of cytochrome c, but the ability of cytochrome c to activate apoptosis is critically dependent upon other signals. PMID- 10364156 TI - Inflammatory mast cells up-regulate angiogenesis during squamous epithelial carcinogenesis. AB - Expression of HPV16 early region genes in basal keratinocytes of transgenic mice elicits a multistage pathway to squamous carcinoma. We report that infiltration by mast cells and activation of the matrix metalloproteinase MMP-9/gelatinase B coincides with the angiogenic switch in premalignant lesions. Mast cells infiltrate hyperplasias, dysplasias, and invasive fronts of carcinomas, but not the core of solid tumors, where they degranulate in close apposition to capillaries and epithelial basement membranes, releasing mast-cell-specific serine proteases MCP-4 (chymase) and MCP-6 (tryptase). MCP-6 is shown to be a mitogen for dermal fibroblasts that proliferate in the reactive stroma, whereas MCP-4 can activate progelatinase B and induce hyperplastic skin to become angiogenic in an in vitro bioassay. Notably, premalignant angiogenesis is abated in a mast-cell-deficient (KITW/KITWWv) HPV16 transgenic mouse. The data indicate that neoplastic progression in this model involves exploitation of an inflammatory response to tissue abnormality. Thus, regulation of angiogenesis during squamous carcinogenesis is biphasic: In hyperplasias, dysplasias, and invading cancer fronts, inflammatory mast cells are conscripted to reorganize stromal architecture and hyperactivate angiogenesis; within the cancer core, upregulation of angiogenesis factors in tumor cells apparently renders them self sufficient at sustaining neovascularization. PMID- 10364157 TI - Direct interaction of hematopoietic transcription factors PU.1 and GATA-1: functional antagonism in erythroid cells. AB - Malignant transformation usually inhibits terminal cell differentiation but the precise mechanisms involved are not understood. PU.1 is a hematopoietic-specific Ets family transcription factor that is required for development of some lymphoid and myeloid lineages. PU.1 can also act as an oncoprotein as activation of its expression in erythroid precursors by proviral insertion or transgenesis causes erythroleukemias in mice. Restoration of terminal differentiation in the mouse erythroleukemia (MEL) cells requires a decline in the level of PU.1, indicating that PU.1 can block erythroid differentiation. Here we investigate the mechanism by which PU.1 interferes with erythroid differentiation. We find that PU.1 interacts directly with GATA-1, a zinc finger transcription factor required for erythroid differentiation. Interaction between PU.1 and GATA-1 requires intact DNA-binding domains in both proteins. PU.1 represses GATA-1-mediated transcriptional activation. Both the DNA binding and transactivation domains of PU.1 are required for repression and both domains are also needed to block terminal differentiation in MEL cells. We also show that ectopic expression of PU.1 in Xenopus embryos is sufficient to block erythropoiesis during normal development. Furthermore, introduction of exogenous GATA-1 in both MEL cells and Xenopus embryos and explants relieves the block to erythroid differentiation imposed by PU.1. Our results indicate that the stoichiometry of directly interacting but opposing transcription factors may be a crucial determinant governing processes of normal differentiation and malignant transformation. PMID- 10364158 TI - Cell cycle-regulated histone acetylation required for expression of the yeast HO gene. AB - Expression of the yeast HO gene in late G1 of the cell cycle requires the SWI/SNF chromatin remodeling complex, the Gcn5p histone acetyltransferase, and two different sequence-specific transcriptional activators, Swi5p and Swi4p/Swi6p. We have used chromatin immunoprecipitation assays to investigate the role of each of these trans-acting factors in establishing a cell cycle-regulated domain of histone acetylation surrounding the HO upstream regulatory region. We detect a approximately 1-kb domain of H3 and H4 acetylation that is established in mid-G1, prior to and independent of HO transcription, which then declines with kinetics similar to inactivation of HO. This cell cycle burst of histone acetylation requires Gcn5p, SWI/SNF, and the Swi5p activator, but occurs in the absence of the Swi4p activator. We also find that inactivation of the Sin3p/Rpd3p deacetylase complex leads to a high level of acetylation at the HO locus throughout the cell cycle. We propose a sequential model for activation of HO in which the Swi5p-dependent recruitment of the Gcn5p acetyltransferase requires chromatin remodeling events by the SWI/SNF complex. PMID- 10364159 TI - Regulation of 4E-BP1 phosphorylation: a novel two-step mechanism. AB - The multisubunit eukaryotic translation initiation factor (eIF) 4F recruits 40S ribosomal subunits to the 5' end of mRNA. The eIF4F subunit eIF4E interacts directly with the mRNA 5' cap structure. Assembly of the eIF4F complex is inhibited by a family of repressor polypeptides, the eIF4E-binding proteins (4E BPs). Binding of the 4E-BPs to eIF4E is regulated by phosphorylation: Hypophosphorylated 4E-BP isoforms interact strongly with eIF4E, whereas hyperphosphorylated isoforms do not. 4E-BP1 is hypophosphorylated in quiescent cells, but is hyperphosphorylated on multiple sites following exposure to a variety of extracellular stimuli. The PI3-kinase/Akt pathway and the kinase FRAP/mTOR signal to 4E-BP1. FRAP/mTOR has been reported to phosphorylate 4E-BP1 directly in vitro. However, it is not known if FRAP/mTOR is responsible for the phosphorylation of all 4E-BP1 sites, nor which sites must be phosphorylated to release 4E-BP1 from eIF4E. To address these questions, a recombinant FRAP/mTOR protein and a FRAP/mTOR immunoprecipitate were utilized in in vitro kinase assays to phosphorylate 4E-BP1. Phosphopeptide mapping of the in vitro-labeled protein yielded two 4E-BP1 phosphopeptides that comigrated with phosphopeptides produced in vivo. Mass spectrometry analysis indicated that these peptides contain phosphorylated Thr-37 and Thr-46. Thr-37 and Thr-46 are efficiently phosphorylated in vitro by FRAP/mTOR when 4E-BP1 is bound to eIF4E. However, phosphorylation at these sites was not associated with a loss of eIF4E binding. Phosphorylated Thr-37 and Thr-46 are detected in all phosphorylated in vivo 4E BP1 isoforms, including those that interact with eIF4E. Finally, mutational analysis demonstrated that phosphorylation of Thr-37/Thr-46 is required for subsequent phosphorylation of several carboxy-terminal serum-sensitive sites. Taken together, our results suggest that 4E-BP1 phosphorylation by FRAP/mTOR on Thr-37 and Thr-46 is a priming event for subsequent phosphorylation of the carboxy-terminal serum-sensitive sites. PMID- 10364160 TI - A PDK1 homolog is necessary and sufficient to transduce AGE-1 PI3 kinase signals that regulate diapause in Caenorhabditis elegans. AB - An insulin receptor-like signaling pathway regulates Caenorhabditis elegans metabolism, development, and longevity. Inactivation of the insulin receptor homolog DAF-2, the AGE-1 PI3K, or the AKT-1 and AKT-2 kinases causes a developmental arrest at the dauer stage. A null mutation in the daf-16 Fork head transcription factor alleviates the requirement for signaling through this pathway. We show here that a loss-of-function mutation in pdk-1, the C. elegans homolog of the mammalian Akt/PKB kinase PDK1, results in constitutive arrest at the dauer stage and increased life span; these phenotypes are suppressed by a loss of function mutation in daf-16. An activating mutation in pdk-1 or overexpression of wild-type pdk-1 relieves the requirement for AGE-1 PI3K signaling. Therefore, pdk-1 activity is both necessary and sufficient to propagate AGE-1 PI3K signals in the DAF-2 insulin receptor-like signaling pathway. The activating mutation in pdk-1 requires akt-1 and akt-2 gene activity in order to suppress the dauer arrest phenotype of age-1. This indicates that the major function of C. elegans PDK1 is to transduce signals from AGE-1 to AKT-1 and AKT-2. The activating pdk-1 mutation is located in a conserved region of the kinase domain; the equivalent amino acid substitution in human PDK1 activates its kinase activity toward mammalian Akt/PKB. PMID- 10364161 TI - Negative regulation of male development in Caenorhabditis elegans by a protein protein interaction between TRA-2A and FEM-3. AB - The tra-2 gene of the nematode Caenorhabditis elegans encodes a predicted membrane protein, TRA-2A, that promotes XX hermaphrodite development. Genetic analysis suggests that tra-2 is a negative regulator of three genes that are required for male development: fem-1, fem-2, and fem-3. We report that the carboxy-terminal region of TRA-2A interacts specifically with FEM-3 in the yeast two-hybrid system and in vitro. Consistent with the idea that FEM-3 is a target of negative regulation, we find that excess FEM-3 can overcome the feminizing effect of tra-2 and cause widespread masculinization of XX somatic tissues. In turn, we show that the masculinizing effects of excess FEM-3 can be suppressed by overproduction of the carboxy-terminal domain of TRA-2A. A FEM-3 fragment that retains TRA-2A-binding activity can masculinize fem-3(+) animals, but not fem-3 mutants, suggesting that it is possible to release and to activate endogenous FEM 3 by titrating TRA-2A. We propose that TRA-2A prevents male development by interacting directly with FEM-3 and that a balance between the opposing activities of TRA-2A and FEM-3 determines sex-specific cell fates in somatic tissues. When the balance favors FEM-3, it acts through or with the other FEM proteins to promote male cell fates. PMID- 10364162 TI - The Drosophila p38 MAPK pathway is required during oogenesis for egg asymmetric development. AB - In mammalian cells, the p38 mitogen-activated protein kinase (MAPK) pathway is activated in response to a variety of environmental stresses and inflammatory stimuli. However, the role of p38 MAPK signaling in unchallenged conditions remains largely unknown. We have isolated mutations in a Drosophila p38 MAPKK gene homolog, licorne (lic), and show that during oogenesis, lic is required in the germ line for correct asymmetric development of the egg. In lic mutant egg chambers, oskar mRNA posterior localization is not properly maintained, resulting in anteroposterior patterning defects in the embryo. Furthermore, lic loss-of function in the germ line leads to reduced EGF receptor activity in dorsal follicle cells and ventralization of the egg shell. Both these defects are associated with a diminution of gurken protein levels in the oocyte. Our phenotypic data argue for a role of lic in a post-transcriptional regulation of the grk gene. Furthermore, they show that in addition to the well-characterized Ras/Raf/ERK MAPK pathway acting in the follicle cells, another related signaling cascade, the p38 MAPK pathway, is required in the germ line for correct axes determination. These results provide the first genetic demonstration of an essential function for a p38 pathway during development. PMID- 10364163 TI - Hrs, a FYVE finger protein localized to early endosomes, is implicated in vesicular traffic and required for ventral folding morphogenesis. AB - Hrs is an early endosomal protein homologous to Vps27p, a yeast protein required for vesicular trafficking. Hrs has a FYVE double zinc finger domain, which specifically binds phosphatidylinositol(3)-phosphate and is conserved in several proteins involved in vesicular traffic. To understand the physiological role of Hrs, we generated mice carrying a null mutation of the gene. Hrs homozygous mutant embryos developed with their ventral region outside of the yolk sac, had two independent bilateral heart tubes (cardia bifida), lacked a foregut, and died around embryonic day 11 (E11). These phenotypes arise from a defect in ventral folding morphogenesis that occurs normally around E8.0. Significant apoptosis was detected in the ventral region of mutant embryos within the definitive endoderm, suggesting an important role of this germ layer in ventral folding morphogenesis. Abnormally enlarged early endosomes were detected in the mutants in several tissues including definitive endoderm, suggesting that a deficiency in vesicular transport via early endosomes underlies the mutant phenotype. The vesicular localization of Hrs was disrupted in cells treated with wortmannin, implicating Hrs in the phosphatidylinositol 3-kinase pathway of membrane trafficking. PMID- 10364165 TI - DNA polymerases: structural diversity and common mechanisms. PMID- 10364164 TI - Periodic repression of Notch pathway genes governs the segmentation of Xenopus embryos. AB - During the development of the vertebrate embryo, genes encoding components of the Notch signaling pathway are required for subdividing the paraxial mesoderm into repeating segmental structures, called somites. These genes are thought to act in the presomitic mesoderm when cells form prospective somites, called somitomeres, but their exact function remains unknown. To address this issue, we have identified two novel genes, called ESR-4 and ESR-5, which are transcriptionally activated in the somitomeres of Xenopus embryos by the Su(H)-dependent Notch signaling pathway. We show that the expression of these genes divides each somitomere into an anterior and posterior half, and that this pattern of expression is generated by a mechanism that actively represses the expression of the Notch pathway genes when paraxial cells enter a critical region and form a somitomere. Repression of Notch signaling during somitomere formation requires a negative feedback loop and inhibiting the activity of genes in this loop has a profound effect on somitomere size. Finally we present evidence that once somitomeres form, ESR-5 mediates a positive feedback loop, which maintains the expression of Notch pathway genes. We propose a model in which Notch signaling plays a key role in both establishing and maintaining segmental identity during somitomere formation in Xenopus embryos. PMID- 10364166 TI - A cell surface ADP-ribosyltransferase modulates T cell receptor association and signaling. AB - ART-1, a cell surface ADP-ribosyltransferase, is imbedded in the membrane by a glycosylphosphatidylinositol anchor. Function of this enzyme in mouse T lymphocytes is to transfer ADP-ribose groups from NAD to arginine residues, exposed on the extracellular domain of cell surface molecules. As a consequence, T cell responses are modulated. To explore the precise action of the enzyme, the T cell lymphoma EL-4 was transfected with the ART-1 gene, and its effects were examined. It is shown that ART-1 ADP-ribosylates distinct cell surface molecules, causing inhibition of T cell receptor signaling, concomitant to suppression of p56(lck) kinase activation. These effects are explained by failure of T cell receptors and co-receptors to associate into a contiguous and functional receptor cluster. PMID- 10364167 TI - Role of adapter function in oncoprotein-mediated activation of NF-kappaB. Human T cell leukemia virus type I Tax interacts directly with IkappaB kinase gamma. AB - Mechanisms by which the human T-cell leukemia virus type I Tax oncoprotein activates NF-kappaB remain incompletely understood. Although others have described an interaction between Tax and a holo-IkappaB kinase (IKK) complex, the exact details of protein-protein contact are not fully defined. Here we show that Tax binds to neither IKK-alpha nor IKK-beta but instead complexes directly with IKK-gamma, a newly characterized component of the IKK complex. This direct interaction with IKK-gamma correlates with Tax-induced IkappaB-alpha phosphorylation and NF-kappaB activation. Thus, our findings establish IKK-gamma as a key molecule for adapting an oncoprotein-specific signaling to IKK-alpha and IKK-beta. PMID- 10364168 TI - Peptidoglycan- and lipoteichoic acid-induced cell activation is mediated by toll like receptor 2. AB - The life-threatening complications of sepsis in humans are elicited by infection with Gram-negative as well as Gram-positive bacteria. Recently, lipopolysaccharide (LPS), a major biologically active agent of Gram-negative bacteria, was shown to mediate cellular activation by a member of the human Toll like receptor family, Toll-like receptor (TLR) 2. Here we investigate the mechanism of cellular activation by soluble peptidoglycan (sPGN) and lipoteichoic acid (LTA), main stimulatory components of Gram-positive bacteria. Like LPS, sPGN and LTA bind to the glycosylphosphatidylinositol-anchored membrane protein CD14 and induce activation of the transcription factor NF-kappaB in host cells like macrophages. We show that whole Gram-positive bacteria, sPGN and LTA induce the activation of NF-kappaB in HEK293 cells expressing TLR2 but not in cells expressing TLR1 or TLR4. The sPGN- and LTA-induced NF-kappaB activation was not inhibited by polymyxin B, an antibiotic that binds and neutralizes LPS. Coexpression together with membrane CD14 enhances sPGN signal transmission through TLR2. In contrast to LPS signaling, activation of TLR2 by sPGN and LTA does not require serum. These findings identify TLR2 as a signal transducer for sPGN and LTA in addition to LPS. PMID- 10364169 TI - Systems properties of the Haemophilus influenzae Rd metabolic genotype. AB - Haemophilus influenzae Rd was the first free-living organism for which the complete genomic sequence was established. The annotated sequence and known biochemical information was used to define the H. influenzae Rd metabolic genotype. This genotype contains 488 metabolic reactions operating on 343 metabolites. The stoichiometric matrix was used to determine the systems characteristics of the metabolic genotype and to assess the metabolic capabilities of H. influenzae. The need to balance cofactor and biosynthetic precursor production during growth on mixed substrates led to the definition of six different optimal metabolic phenotypes arising from the same metabolic genotype, each with different constraining features. The effects of variations in the metabolic genotype were also studied, and it was shown that the H. influenzae Rd metabolic genotype contains redundant functions under defined conditions. We thus show that the synthesis of in silico metabolic genotypes from annotated genome sequences is possible and that systems analysis methods are available that can be used to analyze and interpret phenotypic behavior of such genotypes. PMID- 10364170 TI - Brefeldin A inhibited activity of the sec7 domain of p200, a mammalian guanine nucleotide-exchange protein for ADP-ribosylation factors. AB - A brefeldin A (BFA)-inhibited guanine nucleotide-exchange protein (GEP) for ADP ribosylation factors (ARF) was purified earlier from bovine brain cytosol. Cloning and expression of the cDNA confirmed that the recombinant protein (p200) is a BFA-sensitive ARF GEP. p200 contains a domain that is 50% identical in amino acid sequence to a region in yeast Sec7, termed the Sec7 domain. Sec7 domains have been identified also in other proteins with ARF GEP activity, some of which are not inhibited by BFA. To identify structural elements that influence GEP activity and its BFA sensitivity, several truncated mutants of p200 were made. Deletion of sequence C-terminal to the Sec7 domain did not affect GEP activity. A protein lacking 594 amino acids at the N terminus, as well as sequence following the Sec7 domain, also had high activity. The mutant lacking 630 N-terminal amino acids was, however, only 1% as active, as was the Sec7 domain itself (mutant lacking 697 N-terminal residues). It appears that the Sec7 domain of p200 contains the catalytic site but additional sequence (perhaps especially that between positions 595 and 630) modifies activity dramatically. Myristoylated recombinant ARFs were better than non-myristoylated as substrates; ARFs 1 and 3 were better than ARF5, and no activity was detected with ARF6. Physical interaction of the Sec7 domain with an ARF1 mutant was demonstrated, but it was much weaker than that of the cytohesin-1 Sec7 domain with the same ARF protein. Effects of BFA on p200 and all mutants with high activity were similar with approximately 50% inhibition at 200 kDa) protein and stained primarily the coated pit region of the proximal tubule in a manner similar to that described for megalin (gp330). We then confirmed that both mAb 10A3 and a known anti-megalin mAb immunoprecipitated and immunoblotted the same protein, namely megalin. mAb 10A3 specifically co-precipitated NHE3 but not villin or NaPi-2 from solubilized renal membranes, indicating specificity of the NHE3-megalin interaction. When immunoprecipitations were performed using either 10A3 or anti-NHE3 mAb 2B9 after separation of solubilized renal proteins by sucrose velocity gradient centrifugation, we found that NHE3 exists in two states with distinct sedimentation coefficients, a 9.6 S megalin-free form and a 21 S megalin-bound form, and that when NHE3 assembles with megalin, epitopes within the carboxyl-terminal 131 amino acids of NHE3 are blocked. Taken together, these findings indicate that a significant pool of NHE3 exists as a multimeric complex with megalin in the brush border of the proximal tubule. PMID- 10364183 TI - Accelerated conversion of human plasminogen activator inhibitor-1 to its latent form by antibody binding. AB - The serpin plasminogen activator inhibitor-1 (PAI-1) slowly converts to an inactive latent form by inserting a major part of its reactive center loop (RCL) into its beta-sheet A. A murine monoclonal antibody (MA-33B8), raised against the human plasminogen activator (tPA).PAI-1 complex, rapidly inactivates PAI-1. Results presented here indicate that MA-33B8 induces acceleration of the active to-latent conversion. The antibody-induced inactivation of PAI-1 labeled with the fluorescent probe N, N'-dimethyl-N-(acetyl)-N'-(7-nitrobenz-2-oxa-1,3-diazol-4 yl) ethylene diamine (NBD) at P9 in the RCL caused a fluorescence enhancement and shift identical to those accompanying the spontaneous conversion of the P9.NBD PAI-1 to the latent form. Like latent PAI-1, antibody-inactivated PAI-1 was protected from cleavage by elastase. The rate constants for MA-33B8 binding, measured by NBD fluorescence or inactivation, were similar (1.3-1.8 x 10(4) M-1 s 1), resulting in a 4000-fold faster inactivation at 4.2 microM antibody binding sites. The apparent antibody binding rate constant, at least 1000 times slower than one limited by diffusion, indicates that exposure of its epitope depends on an unfavorable equilibrium of PAI-1. Our observations are consistent with this idea and suggest that the equilibrium involves partial insertion of the RCL into sheet A: latent, RCL-cleaved, and tPA-complexed PAI-1, which are inactive loop inserted forms, bound much faster than active PAI-1 to MA-33B8, whereas two loop extracted forms of PAI-1, modified to prevent loop insertion, did not bind or bound much more weakly to the antibody. PMID- 10364185 TI - The importance of ATP binding and hydrolysis by hsp90 in formation and function of protein heterocomplexes. AB - The chaperone hsp90 is capable of binding and hydrolyzing ATP. Using information on a related ATPase, DNA gyrase B, we selected three conserved residues in hsp90's ATP-binding domain for mutation. Two of these mutations eliminate nucleotide binding, while the third retains nucleotide binding but is apparently deficient in ATP hydrolysis. We first analyzed how these mutations affect hsp90's binding to the co-chaperones p23 and Hop, and to the hydrophobic resin, phenyl Sepharose. These experiments showed that ATP's effects, specifically, increased affinity for p23 and decreased affinity for Hop and phenyl-Sepharose, are brought on by ATP binding alone. We also tested the ability of hsp90 mutants to assist hsp70, hsp40, and Hop in the refolding of denatured firefly luciferase. While hsp90 is capable of participating in this process in a nucleotide-independent manner, the ability to hydrolyze ATP markedly potentiates hsp90's effect. Finally, we assembled progesterone receptor heterocomplexes with hsp70, hsp40, Hop, p23, and wild type or mutant hsp90. While neither ATP binding nor hydrolysis was necessary to bind hsp90 to the receptor, mature complexes containing p23 and capable of hormone binding were only obtained with wild type hsp90. PMID- 10364186 TI - Respiratory uncoupling induces delta-aminolevulinate synthase expression through a nuclear respiratory factor-1-dependent mechanism in HeLa cells. AB - Nuclear respiratory factor (NRF)-1 appears to be important for the expression of several respiratory genes, but there is no direct evidence that NRF-1 transduces a physiological signal into the production of an enzyme critical for mitochondrial biogenesis. We generated HeLa cells containing plasmids allowing doxycycline-inducible expression of uncoupling protein (UCP)-1. In the absence of doxycycline, UCP-1 mRNA and protein were undetectable. In the presence of doxycycline, UCP-1 was expressed and oxygen consumption doubled. This rise in oxygen consumption was associated with an increase in NRF-1 mRNA. It was also associated with an increase in NRF-1 protein binding activity as determined by electrophoretic mobility shift assay using a functional NRF-1 binding site from the delta-aminolevulinate (ALA) synthase promoter. Respiratory uncoupling also caused a time-dependent increase in protein levels of ALA synthase, an early marker for mitochondrial biogenesis. ALA synthase induction by respiratory uncoupling was prevented by transfecting cells with an oligonucleotide antisense to the region of the NRF-1 initiation codon; a scrambled oligonucleotide with the same base composition had no effect. Respiratory uncoupling increases oxygen consumption and lowers energy reserves. In HeLa cells, uncoupling also increases ALA synthase, an enzyme critical for mitochondrial respiration, but only if translatable mRNA for NRF-1 is available. These data suggest that the transcription factor NRF-1 plays a key role in cellular adaptation to energy demands by translating physiological signals into an increased capacity for generating energy. PMID- 10364188 TI - Role of residues 311/312 in actin-tropomyosin interaction. In vitro motility study using yeast actin mutant e311a/r312a. AB - According to the Lorenz et al. (Lorenz, M., Poole, K. J., Popp, D., Rosenbaum, G., and Holmes, K. C. (1995) J. Mol. Biol. 246, 108-119) atomic model of the actin-tropomyosin complex, actin residue Asp-311 (Glu-311 in yeast) is predicted to have a high binding energy contribution to actin-tropomyosin binding. Using the yeast actin mutant E311A/R312A in the in vitro motility assays, we have investigated the role of these residues in such interactions. Wild type (wt) yeast actin, like skeletal alpha-actin, is fully regulated when complexed with tropomyosin (Tm) and troponin (Tn). Structure-function comparisons of the wt and E311A/R312A actins show no significant differences between them, and the unregulated F-actins slide at similar speeds in the in vitro motility assay. However, in the presence of Tm and Tn, the mutation increases both the sliding speed and the number of moving filaments at high pCa values, shifting the speed pCa curve nearly 0.5 pCa units to the left. Tm alone (no Tn) inhibits the motilities of both actins at low heavy meromyosin densities but potentiates only the motility of the mutant actin at high heavy meromyosin densities. Actin-Tm binding measurements indicate no significant difference between wt and E311A/R312A actin in Tm binding. These results implicate allosteric effects in the regulation of actomyosin function by tropomyosin. PMID- 10364187 TI - Novel form of lipolysis induced by leptin. AB - Hyperleptinemia causes disappearance of body fat without a rise in free fatty acids (FFA) or ketones, suggesting that leptin can deplete adipocytes of fat without releasing FFA. To test this, we measured FFA and glycerol released from adipocytes obtained from normal lean Zucker diabetic fatty rats (+/+) and incubated for 0, 3, 6, or 24 h in either 20 ng/ml recombinant leptin or 100 nM norepinephrine (NE). Whereas NE increased both FFA and glycerol release from adipocytes of +/+ rats, leptin increased glycerol release in +/+ adipocytes without a parallel increase in FFA release. In adipocytes of obese Zucker diabetic fatty rats (fa/fa) with defective leptin receptors, NE increased both FFA and glycerol release, but leptin had no effect on either. Leptin significantly lowered the mRNA of leptin and fatty acid synthase of adipocytes (FAS) (p < 0.05), and up-regulated the mRNA of peroxisome proliferator-activated receptor (PPAR)-alpha, carnitine palmitoyl transferase-1, (CPT-1), and acyl CoA oxidase (ACO) (p < 0.05). NE (100 nM) also lowered leptin mRNA (p < 0.05) but did not affect FAS, PPARalpha, ACO, or CPT-1 expression. We conclude that in normal adipocytes leptin directly decreases FAS expression, increases PPARalpha and the enzymes of FFA oxidation, and stimulates a novel form of lipolysis in which glycerol is released without a proportional release of FFA. PMID- 10364189 TI - Molecular chaperones stimulate the functional expression of the cocaine-sensitive serotonin transporter. AB - The serotonin transporter (SERT) is an N-glycosylated integral membrane protein that is predicted to contain 12 transmembrane regions. SERT is the major binding site in the brain for antidepressant drugs, and it also binds amphetamines and cocaine. The ability of various molecular chaperones to interact with a tagged version of SERT (Myc-SERT) was investigated using the baculovirus expression system. Overexpression of Myc-SERT using the baculovirus system led to substantial quantities of inactive transporter, together with small amounts of fully active and, therefore, correctly folded molecules. The high levels of inactive Myc-SERT probably arose because folding was rate-limiting due, perhaps, to insufficient molecular chaperones. Therefore, Myc-SERT was co-expressed with the endoplasmic reticulum (ER) molecular chaperones calnexin, calreticulin and immunoglobulin heavy chain binding protein (BiP), and the foldase, ERp57. The expression of functional Myc-SERT, as determined by an inhibitor binding assay, was enhanced nearly 3-fold by co-expressing calnexin, and to a lesser degree on co-expression of calreticulin and BiP. Co-expression of ERp57 did not increase the functional expression of Myc-SERT. A physical interaction between Myc-SERT calnexin and Myc-SERT-calreticulin was demonstrated by co-immunoprecipitation. These associations were inhibited in vivo by deoxynojirimycin, an inhibitor of N glycan precusor trimming that is known to prevent the calnexin/calreticulin-N glycan interaction. Functional expression of the unglycosylated SERT mutant, SERT QQ, was also increased on co-expression of calnexin, suggesting that the interaction between calnexin and SERT is not entirely dictated by the N-glycan. SERT is the first member of the neurotransmitter transporter family whose folding has been shown to be assisted by the molecular chaperones calnexin, calreticulin, and BiP. PMID- 10364190 TI - Characterization of mouse nNOS2, a natural variant of neuronal nitric-oxide synthase produced in the central nervous system by selective alternative splicing. AB - Mouse neuronal nitric-oxide synthase 2 (nNOS2) is a unique natural variant of constitutive neuronal nitric-oxide synthase (nNOS) specifically expressed in the central nervous system having a 105-amino acid deletion in the heme-binding domain as a result of in-frame mutation by specific alternative splicing. The mouse nNOS2 cDNA gene was heterologously expressed in Escherichia coli, and the resultant product was characterized spectroscopically in detail. Purified recombinant nNOS2 contained heme but showed no L-arginine- and NADPH-dependent citrulline-forming activity in the presence of Ca2+-promoted calmodulin, elicited a sharp electron paramagnetic resonance (EPR) signal at g = 6.0 indicating the presence of a high spin ferriheme as isolated and showed a peak at around 420 nm in the CO difference spectrum, instead of a 443-nm peak detected with the recombinant wild-type nNOS1 enzyme. Thus, although the heme domain of nNOS2 is capable of binding heme, the heme coordination geometry is highly abnormal in that it probably has a proximal non-cysteine thiolate ligand both in the ferric and ferrous states. Moreover, negligible spectral perturbation of the nNOS2 ferriheme was detected upon addition of either L-arginine or imidazole. These provide a possible rational explanation for the inability of nNOS2 to catalyze the cytochrome P450-type monooxygenase reaction. PMID- 10364191 TI - Potent and stable attenuation of live-HIV-1 by gain of a proteolysis-resistant inhibitor of NF-kappaB (IkappaB-alphaS32/36A) and the implications for vaccine development. AB - Live-attenuated human immunodeficiency viruses (HIVs) are candidates for Acquired Immunodeficiency Syndrome (AIDS) vaccine. Based on the simian immunodeficiency virus (SIV) model for AIDS, loss-of-function (e.g. deletion of accessory genes such as nef) has been forwarded as a primary approach for creating enfeebled, but replication-competent, HIV-1/SIV. Regrettably, recent evidence suggests that loss of-function alone is not always sufficient to prevent the emergence of virulent mutants. New strategies that attenuate via mechanisms distinct from loss-of function are needed for enhancing the safety phenotype of viral genome. Here, we propose gain-of-function to be used simultaneously with loss-of-function as a novel approach for attenuating HIV-1. We have constructed an HIV-1 genome carrying the cDNA of a proteolysis-resistant nuclear factor-kappaB inhibitor (IkappaB-alphaS32/36A) in the nef region. HIV-1 expressing IkappaB-alphaS32/36A down-regulates viral expression and is highly attenuated in both Jurkat and peripheral blood mononuclear cells. We provide formal proof that the phenotypic and attenuating characteristics of IkappaB-alphaS32/36A permit its stable maintenance in a live, replicating HIV-1 despite 180 days of forced ex vivo passaging in tissue culture. As compared with other open-reading frames embedded into HIV/SIV genome, this degree of stability is unprecedented. Thus, IkappaB alphaS32/36A offers proof-of-principle that artifactually gained functions, when used to attenuate the replication of live HIV-1, can be stable. These findings illustrate gain-of-function as a feasible strategy for developing safer live attenuated HIVs to be tested as candidates for AIDS vaccine. PMID- 10364192 TI - Backbone cyclic peptide antagonists, derived from the insect pheromone biosynthesis activating neuropeptide, inhibit sex pheromone biosynthesis in moths. AB - We describe an application of the backbone cyclization and cycloscan concept for the design and synthesis of pheromone biosynthesis activating neuropeptide (PBAN) antagonists capable of inhibiting sex pheromone biosynthesis in Heliothis peltigera female moths. Two backbone cyclic (BBC) sub-libraries were designed and synthesized. The structure of the first sub-library ([Arg27]PBAN27-33NH2, termed the Ser sub-library) was based on the active C-terminal hexapeptide sequence (Tyr Phe-Ser-Pro-Arg-Leu-NH2) of PBAN1-33NH2, which was found to comprise its active core. The second sub-library ([Arg27, D-Phe30]PBAN27-33NH2, termed the D-Phe sub library) was based on the sequence of the lead antagonist Arg-Tyr-Phe-(D)Phe-Pro Arg-Leu-NH2. In both sub-libraries the Pro residue was replaced by an Nalpha(omega-amino-alkyl)Gly building unit having various lengths of the alkyl chain. All the cyclic peptides in each sub-library had the same primary sequence and the same location of the ring. The members of each library differed from each other by the bridge size and bridge chemistry. Screening of the two libraries for pheromonotropic antagonists resulted in the disclosure of four compounds that fully inhibited sex pheromone biosynthesis at 1 nmol and were devoid of agonistic activity. All antagonistic peptides originated from the D-Phe sub-library. Substitution of the D-Phe30 amino acid with a Ser resulted in a loss of antagonistic activity. Agonistic activities were exhibited by peptides from both sub-libraries. PMID- 10364193 TI - Oxidative stress triggers STAT3 tyrosine phosphorylation and nuclear translocation in human lymphocytes. AB - Oxidizing agents are powerful activators of factors responsible for the transcriptional activation of cytokine-encoding genes involved in tissue injury. In this study we show evidence that STAT3 is a transcription factor whose activity is modulated by H2O2 in human lymphocytes, in which endogenous catalase had previously been inhibited. H2O2-induced nuclear translocation of STAT3 to form sequence-specific DNA-bound complexes was evidenced by immunoblotting of nuclear fractions and electrophoretic mobility shift assays, and vanadate was found to strongly synergize with H2O2. Moreover, anti-STAT3 antibodies specifically precipitated a protein of 92 kDa that becomes phosphorylated on tyrosine upon lymphocyte treatment with H2O2. Phenylarsine oxide, a tyrosine phosphatase inhibitor, and genistein, a tyrosine kinase inhibitor, cooperated and cancelled, respectively, the H2O2-promoted STAT3 nuclear translocation. Evidence is also presented, using Fe2+/Cu2+ ions, that.OH generated from H2O2 through Fenton reactions could be a candidate oxygen reactive species to directly activate STAT3. Present data suggest that H2O2 and vanadate are likely to inhibit the activity of intracellular tyrosine phosphatase(s), leading to enhanced STAT3 tyrosine phosphorylation and hence its translocation to the nucleus. These results demonstrate that the DNA binding activity of STAT3 can be modulated by oxidizing agents and provide a framework to understand the effects of oxidative stress on the JAK-STAT signaling pathway. PMID- 10364194 TI - Identification of the G protein-activating domain of the natriuretic peptide clearance receptor (NPR-C). AB - We have shown recently that the 37-amino acid intracellular domain of the single transmembrane, natriuretic peptide clearance receptor, NPR-C, which is devoid of kinase and guanylyl cyclase activities, activates selectively Gi1 and Gi2 in gastric and tenia coli smooth muscle. In this study, we have used synthetic peptide fragments of the N-terminal, C-terminal, and middle regions of the cytoplasmic domain of NPR-C to identify the G protein-activating sequence. A 17 amino acid peptide of the middle region (Arg469-Arg485), denoted Peptide 4, which possesses two N-terminal arginine residues and a C-terminal B-B-X-X-B motif (where B and X are basic and non-basic residues, respectively) bound selectively to Gi1 and Gi2, activated phospholipase C-beta3 via the betagamma subunits, inhibited adenylyl cyclase, and induced smooth muscle contraction, in similar fashion to the selective NPR-C ligand, cANP4-23. A similar sequence (Peptide 3), but with a partial C-terminal motif, had minimal activity. Sequences which possessed either the N-terminal basic residues (Peptide 1) or the C-terminal B-B X-X-B motif (Peptide 2) were inactive. Peptide 2, however, inhibited G protein activation and cellular responses mediated by the stimulatory Peptide 4 and by cANP4-23, suggesting that the B-B-X-X-B motif mediated binding but not activation of G protein, thus causing Peptide 2 to act as a competitive inhibitor of G protein activation. PMID- 10364195 TI - Evidence that MgATP accelerates primary electron transfer in a Clostridium pasteurianum Fe protein-Azotobacter vinelandii MoFe protein nitrogenase tight complex. AB - The nitrogenase catalytic cycle involves binding of the iron (Fe) protein to the molybdenum-iron (MoFe) protein, transfer of a single electron from the Fe protein to the MoFe protein concomitant with the hydrolysis of at least two MgATP molecules, followed by dissociation of the two proteins. Earlier studies found that combining the Fe protein isolated from the bacterium Clostridium pasteurianum with the MoFe protein isolated from the bacterium Azotobacter vinelandii resulted in an inactive, nondissociating Fe protein-MoFe protein complex. In the present work, it is demonstrated that primary electron transfer occurs within this nitrogenase tight complex in the absence of MgATP (apparent first-order rate constant k = 0.007 s-1) and that MgATP accelerates this electron transfer reaction by more than 10,000-fold to rates comparable to those observed within homologous nitrogenase complexes (k = 100 s-1). Electron transfer reactions were confirmed by EPR spectroscopy. Finally, the midpoint potentials (Em) for the Fe protein [4Fe-4S]2+/+ cluster and the MoFe protein P2+/N cluster were determined for both the uncomplexed and complexed proteins and with or without MgADP. Calculations from electron transfer theory indicate that the measured changes in Em are not likely to be sufficient to account for the observed nucleotide-dependent rate accelerations for electron transfer. PMID- 10364196 TI - Tip60 is a nuclear hormone receptor coactivator. AB - The androgen receptor (AR) is a member of the nuclear hormone receptor superfamily. Recent work in this field has been focused upon defining the mechanisms of transcriptional control exacted by members of this superfamily. Using a COOH-terminal region of the human AR in a yeast two-hybrid screen, we have identified Tip60 as an AR-interacting protein. In this report, we show that Tip60, which was originally identified as a coactivator for the human immunodeficiency virus TAT protein, can enhance AR-mediated transactivation in a ligand-dependent manner in LNCaP and COS-1 cell lines. In addition, our experiments show that Tip60 can also enhance transactivation through the estrogen receptor and progesterone receptor in a ligand-dependent manner; thus identifying Tip60 as a nuclear hormone receptor coactivator. Our studies also demonstrate that Tip60 co-immunoprecipitates with the full-length AR in vitro and that, in our system, Tip60 enhances transactivation to levels observed with the coactivators steroid receptor coactivator 1, p300, and CREB-binding protein. The importance of such proteins in enhancing nuclear hormone receptor-mediated transcriptional activation is widely accepted, and this work suggests that Tip60 may have an equally important role to play. PMID- 10364197 TI - Correction of large mispaired DNA loops by extracts of Saccharomyces cerevisiae. AB - Single base mispairs and small loops are corrected by DNA mismatch repair, but little is known about the correction of large loops. In this paper, large loop repair was examined in nuclear extracts of yeast. Biochemical assays showed that repair activity occurred on loops of 16, 27, and 216 bases, whereas a G-T mispair and an 8-base loop were poorly corrected under these conditions. Two modes of loop repair were revealed by comparison of heteroduplexes that contained a site specific nick or were covalently closed. A nick-stimulated repair mode directs correction to the discontinuous strand, regardless of which strand contains the loop. An alternative mode is nick-independent and preferentially removes the loop. Both outcomes of repair were largely eliminated when DNA replication was inhibited, suggesting a requirement for repair synthesis. Excision tracts of 100 200 nucleotides, spanning the position of the loop, were observed on each strand under conditions of limited DNA repair synthesis. Both repair modes were independent of the mismatch correction genes MSH2, MSH3, MLH1, and PMS1, as judged by activity in mutant extracts. Together the loop specificity and mutant results furnish evidence for a large loop repair pathway in yeast that is distinct from mismatch repair. PMID- 10364198 TI - Epidermal growth factor protects epithelial cells against Fas-induced apoptosis. Requirement for Akt activation. AB - Chemotherapeutic drugs that damage DNA kill tumor cells, in part, by inducing the expression of a death receptor such as Fas or its ligand, FasL. Here, we demonstrate that epidermal growth factor (EGF) stimulation of T47D breast adenocarcinoma and embryonic kidney epithelial (HEK293) cells protects these cells from Fas-induced apoptosis. EGF stimulation of epithelial cells also inhibited Fas-induced caspase activation and the proteolysis of signaling proteins downstream of the EGF receptor, Cbl and Akt/protein kinase B (Akt). EGF stimulation of Akt kinase activity blocked Fas-induced apoptosis. Expression of activated Akt in MCF-7 breast adenocarcinoma cells was sufficient to block Fas mediated apoptosis. Inhibition of EGF-stimulated extracellular signal-regulated kinase (ERK) activity did not affect EGF protection from Fas-mediated apoptosis. The findings indicate that EGF receptor stimulation of epithelial cells has a significant survival function against death receptor-induced apoptosis mediated by Akt. PMID- 10364199 TI - Evidence for a role for ADP-ribosylation factor 6 in insulin-stimulated glucose transporter-4 (GLUT4) trafficking in 3T3-L1 adipocytes. AB - ADP-ribosylation factors (ARFs) play important roles in both constitutive and regulated membrane trafficking to the plasma membrane in other cells. Here we have examined their role in insulin-stimulated GLUT4 translocation in 3T3-L1 adipocytes. These cells express ARF5 and ARF6. ARF5 was identified in the soluble protein and intracellular membranes; in response to insulin some ARF5 was observed to re-locate to the plasma membrane. In contrast, ARF6 was predominantly localized to the plasma membrane and did not redistribute in response to insulin. We employed myristoylated peptides corresponding to the NH2 termini of ARF5 and ARF6 to investigate the function of these proteins. Myr-ARF6 peptide inhibited insulin-stimulated glucose transport and GLUT4 translocation by approximately 50% in permeabilized adipocytes. In contrast, myr-ARF1 and myr-ARF5 peptides were without effect. Myr-ARF5 peptide also inhibited the insulin stimulated increase in cell surface levels of GLUT1 and transferrin receptors. Myr-ARF6 peptide significantly decreased cell surface levels of these proteins in both basal and insulin-stimulated states, but did not inhibit the fold increase in response to insulin. These data suggest an important role for ARF6 in regulating cell surface levels of GLUT4 in adipocytes, and argue for a role for both ARF5 and ARF6 in the regulation of membrane trafficking to the plasma membrane. PMID- 10364200 TI - Factors involved in GLUT-1 glucose transporter gene transcription in cardiac muscle. AB - Glucose constitutes a major fuel for the heart, and high glucose uptake during fetal development is coincident with the highest level of expression of the glucose transporter GLUT-1 during life. We have previously reported that GLUT-1 is repressed perinatally in rat heart, and GLUT-4, which shows a low level of expression in the fetal stage, becomes the main glucose transporter in the adult. Here, we show that the perinatal expression of GLUT-1 and GLUT-4 glucose transporters in heart is controlled directly at the level of gene transcription. Transient transfection assays show that the -99/-33 fragment of the GLUT-1 gene is sufficient to drive transcriptional activity in rat neonatal cardiomyocytes. Electrophoretic mobility shift assays demonstrate that the transcription factor Sp1, a trans-activator of GLUT-1 promoter, binds to the -102/-82 region of GLUT-1 promoter during the fetal state but not during adulthood. Mutation of the Sp1 site in this region demonstrates that Sp1 is essential for maintaining a high transcriptional activity in cardiac myocytes. Sp1 is markedly down-regulated both in heart and in skeletal muscle during neonatal life, suggesting an active role for Sp1 in the regulation of GLUT-1 transcription. In all, these results indicate that the expression of GLUT-1 and GLUT-4 in heart during perinatal development is largely controlled at a transcriptional level by mechanisms that might be related to hyperplasia and that are independent from the signals that trigger cell hypertrophy in the developing heart. Furthermore, our results provide the first functional insight into the mechanisms regulating muscle GLUT-1 gene expression in a live animal. PMID- 10364201 TI - Calnexin interaction with N-glycosylation mutants of a polytopic membrane glycoprotein, the human erythrocyte anion exchanger 1 (band 3). AB - The interaction of the endoplasmic reticulum chaperone calnexin with N glycosylation mutants of a polytopic membrane glycoprotein, the human erythrocyte anion exchanger (AE1), was characterized by cell-free translation and in transfected HEK293 cells, followed by co-immunoprecipitation using anti-calnexin antibody. AE1 contains 12-14 transmembrane segments and has a single site of N glycosylation at Asn-642 in the fourth extracytosolic loop. This site was mutated (N642D) to create a nonglycosylated protein. Calnexin showed a preferential interaction with N-glycosylated AE1 relative to nonglycosylated AE1 both in vitro and in vivo. This interaction could be blocked by inhibition of glucosidases I and II with castanospermine. Calnexin had access to novel N-glycosylated sites created in other extracytosolic loops in AE1 by site-directed or insertional mutagenesis. The interaction with AE1 was enhanced when multiple sites were introduced into the same loop or into two different loops. An association of calnexin with truncated versions of N-glycosylated AE1 was detected after release of the nascent chains from ribosomes with puromycin. The results show that the interaction of calnexin with the polytopic membrane glycoprotein AE1 was dependent on the presence but not the location of the oligosaccharide. Furthermore, calnexin was associated with AE1 after release of AE1 from the translocation machinery. PMID- 10364202 TI - D-site binding protein transactivation requires the proline- and acid-rich domain and involves the coactivator p300. AB - The D-site binding protein (DBP) is a member of the proline- and acid-rich (PAR) domain subfamily of basic/leucine zipper proteins and is involved in transcriptional regulation in the liver. Deletion analysis of the DBP protein was carried out in an effort to define the function of the conserved PAR domain. Internal deletions of the protein, i.e. removing portions of the PAR domain, resulted in a substantial loss in transactivation of a high affinity DBP reporter construct when assayed in Hep G2 cells. These same sequences conferred significant transactivation to GAL4 DNA binding domain fusion proteins, indicating that this region acts as part of an independent activation domain comprised of sequences in both the amino terminus and in the PAR domain of DBP. The coexpression of full-length expression constructs for both DBP and hepatic leukemia factor resulted in a dramatic increase in activation mediated by the GAL4-DBP fusion proteins, suggesting the involvement of a regulated coactivator in this process. DBP transactivation appears to be a p300-dependent process, as a 12 S E1A expression construct disrupted DBP-mediated transactivation, and a p300 expression vector, but not a CREB binding protein vector, was able to restore DBP transactivation. These results suggest that the PAR domain is required for DBP activation, which occurs through a regulated, p300-dependent process. PMID- 10364203 TI - Ligand-mediated tertiary structure changes of reconstituted P-glycoprotein. A tryptophan fluorescence quenching analysis. AB - Ligand-dependent changes in accessibility of purified P-glycoprotein, functionally reconstituted in liposomes, were investigated by fluorescence measurements. Trp quenching experiments provided evidence that P-glycoprotein adopts different tertiary structures upon binding of drug substrates in the absence and presence of MgATP and its nonhydrolyzable analog, MgATPgammaS. Five anthracycline derivatives were tested as drug substrates: daunorubicin, 4'-epi doxorubicin, iododoxorubicin, 4-demethoxy-daunorubicin, and methoxy-morpholino doxorubicin. Among them, daunorubicin and 4'-epi-doxorubicin have been shown to be rejected outside the multidrug-resistant cells, whereas the three others have been shown to accumulate in multidrug-resistant cells overexpressing P glycoprotein and therefore retain their cytotoxic activity. A small conformational change was associated with nucleotide binding and amplified after nucleotide hydrolysis. Different conformational states were adopted by P glycoprotein upon the addition of the anthracycline derivatives in the absence and presence of MgATP or MgATPgammaS. These conformational changes are shown to be related to the nature of the antitumor agents and more precisely to their capacity to accumulate in resistant cells. These data also suggest that the cytotoxicity of iododoxorubicin and 4-demethoxy-daunorubicin is related to the fact they are not transported by P-glycoprotein. On the contrary, methoxy morpholino-doxorubicin cytotoxicity may be explained in terms of its rapid reincorporation into the plasma membrane after being transported by P glycoprotein. PMID- 10364204 TI - Protonation/deprotonation reactions triggered by photoactivation of photoactive yellow protein from Ectothiorhodospira halophila. AB - Light-dependent pH changes were measured in unbuffered solutions of wild type photoactive yellow protein (PYP) and its H108F and E46Q variants, using two independent techniques: transient absorption changes of added pH indicator dyes and direct readings with a combination pH electrode. Depending on the absolute pH of the sample, a reversible protonation as well as a deprotonation can be observed upon formation of the transient, blue-shifted photocycle intermediate (pB) of this photoreceptor protein. The latter is observed at very alkaline pH, the former at acidic pH values. At neutral pH, however, the formation of the pB state is not paralleled by significant protonation/deprotonation of PYP, as expected for concomitant protonation of the chromophore and deprotonation of Glu 46 during pB formation. We interpret these results as further evidence that a proton is transferred from Glu-46 to the coumaric acid chromophore of PYP, during pB formation. One cannot exclude the possibility, however, that this transfer proceeds through the bulk aqueous phase. Simultaneously, an amino acid side chain(s) (e.g. His-108) changes from a buried to an exposed position. These results, therefore, further support the idea that PYP significantly unfolds in the pB state and resolve the controversy regarding proton transfer during the PYP photocycle. PMID- 10364205 TI - Structure and evolution of the alternatively spliced fast troponin T isoform gene. AB - The vertebrate fast skeletal muscle troponin T gene, TnTf, produces a complexity of isoforms through differential mRNA splicing. The mechanisms that regulate splicing and the physiological significance of TnTf isoforms are poorly understood. To investigate these questions, we have determined the complete sequence structure of the quail TnTf gene, and we have characterized the developmental expression of alternatively spliced TnTf mRNAs in quail embryonic muscles. We report the following: 1) the quail TnTf gene is significantly larger than the rat TnTf gene and has 8 non-homologous exons, including a pectoral muscle-specific set of alternatively spliced exons; 2) specific sequences are implicated in regulated exon splicing; 3) a 900-base pair sequence element, composed primarily of intron sequence flanking the pectoral muscle-specific exons, is tandemly repeated 4 times and once partially, providing direct evidence that the pectoral-specific TnT exon domain arose by intragenic duplications; 4) a chicken repeat 1 retrotransposon element resides upstream of this repeated intronic/pectoral exon sequence domain and is implicated in transposition of this element into an ancestral genome; and 5) a large set of novel isoforms, produced by regulated exon splicing, is expressed in quail muscles, providing insights into the developmental regulation, physiological function, and evolution of the vertebrate TnTf isoforms. PMID- 10364206 TI - Only multimeric hensin located in the extracellular matrix can induce apical endocytosis and reverse the polarity of intercalated cells. AB - When an intercalated epithelial cell line was seeded at low density and allowed to reach confluence, it located the anion exchanger band 3 in the apical membrane and an H+-ATPase in the basolateral membrane. The same clonal cells seeded at high density targeted these proteins to the reverse location. Furthermore, high density cells had vigorous apical endocytosis, and low density cells had none. The extracellular matrix of high density cells was capable of inducing apical endocytosis and relocation of band 3 to the basolateral membrane in low density cells. A 230-kDa extracellular matrix (ECM) protein termed hensin, when purified to near-homogeneity, was able to reverse the phenotype of the low density cells. Antibodies to hensin prevented this effect, indicating that hensin is necessary for conversion of polarity. We show here that hensin was synthesized by both low density and high density cells. Whereas both phenotypes secreted soluble hensin into their media, only high density cells localized it in their ECM. Analysis of soluble hensin by sucrose density gradients showed that low density cells secreted monomeric hensin, and high density cells secreted higher order multimers. When 35S-labeled monomeric hensin was added to high density cells, they induced its aggregation suggesting that the multimerization was catalyzed by surface events in the high density cells. Soluble monomeric or multimeric hensin did not induce apical endocytosis in low density cells, whereas the more polymerized hensin isolated from insoluble ECM readily induced it. These multimers could be disaggregated by sulfhydryl reagents and by dimethylmaleic anhydride, and treatment of high density ECM by these reagents prevented the induction of endocytosis. These results demonstrate that hensin, like several ECM proteins, needs to be precipitated in the ECM to be functional. PMID- 10364207 TI - Ded1p, a DEAD-box protein required for translation initiation in Saccharomyces cerevisiae, is an RNA helicase. AB - The Ded1 protein (Ded1p), a member of the DEAD-box family, has recently been shown to be essential for translation initiation in Saccharomyces cerevisiae. Here, we show that Ded1p purified from Escherichia coli has an ATPase activity, which is stimulated by various RNA substrates. Using an RNA strand-displacement assay, we show that Ded1p has also an ATP-dependent RNA unwinding activity. Hydrolysis of ATP is required for this activity: the replacement of ATP by a nonhydrolyzable analog or a mutation in the DEAD motif abolishing ATPase activity results in loss of RNA unwinding. We find that cells harboring a Ded1 protein with this mutated DEAD motif are nonviable, suggesting that the ATPase and RNA helicase activities of this protein are essential to the cell. Finally, RNA binding measurements indicate that the presence of ATP, but not ADP, increases the affinity of Ded1p for duplex versus single-stranded RNA; we discuss how this differential effect might drive the unwinding reaction. PMID- 10364208 TI - Alternate coupling of receptors to Gs and Gi in pancreatic and submandibular gland cells. AB - Many Gs-coupled receptors can activate both cAMP and Ca2+ signaling pathways. Three mechanisms for dual activation have been proposed. One is receptor coupling to both Gs and G15 (a Gq class heterotrimeric G protein) to initiate independent signaling cascades that elevate intracellular levels of cAMP and Ca+2, respectively. The other two mechanisms involve cAMP-dependent protein kinase mediated activation of phospholipase Cbeta either directly or by switching receptor coupling from Gs to Gi. These mechanisms were primarily inferred from studies with transfected cell lines. In native cells we found that two Gs-coupled receptors (the vasoactive intestinal peptide and beta-adrenergic receptors) in pancreatic acinar and submandibular gland duct cells, respectively, evoke a Ca2+ signal by a mechanism involving both Gs and Gi. This inference was based on the inhibitory action of antibodies specific for Galphas, Galphai, and phosphatidylinositol 4,5-bisphosphate, pertussis toxin, RGS4, a fragment of beta adrenergic receptor kinase and inhibitors of cAMP-dependent protein kinase. By contrast, Ca2+ signaling evoked by Gs-coupled receptor agonists was not blocked by Gq class-specific antibodies and was unaffected in Galpha15 -/- knockout mice. We conclude that sequential activation of Gs and Gi, mediated by cAMP-dependent protein kinase, may represent a general mechanism in native cells for dual stimulation of signaling pathways by Gs-coupled receptors. PMID- 10364209 TI - Phosphorylation of the myosin-II light chain does not regulate the timing of cytokinesis in fission yeast. AB - Proper coordination of cytokinesis with chromosome separation during mitosis is crucial to ensure that each daughter cell inherits an equivalent set of chromosomes. It has been proposed that one mechanism by which this is achieved is through temporally regulated myosin regulatory light chain (RLC) phosphorylation (Satterwhite, L. L., and Pollard, T. D. (1992) Curr. Opin. Cell Biol. 4, 43-52). A variety of evidence is consistent with this model. A direct test of the importance of RLC phosphorylation in vivo has been done only in Dictyostelium and Drosophila; phosphorylation of the RLC is essential in Drosophila (Jordan, P., and Karess, R. (1997) J. Cell Biol. 139, 1805-1819) but not essential in Dictyostelium (Ostrow, B. D., Chen, P., and Chisholm, R. L. (1994) J. Cell Biol. 127, 1945-1955). The Schizosaccharomyces pombe myosin light chain Cdc4p is essential for cytokinesis, but it was unknown whether phosphorylation played a role in its regulation. Here we show that the S. pombe myosin light chain Cdc4p is phosphorylated in vivo on either serine 2 or 6 but not both. Mutation of either or both of these sites to alanine did not effect the ability of Cdc4p to bind the type II myosin Myo2p, and cells expressing only these mutated versions of Cdc4p grew and divided normally. Similarly, mutation of Ser-2, Ser-6, or both residues to aspartic acid did not affect growth or division of cells. Thus we conclude that phosphorylation of Cdc4p is not essential in vivo for the function of the protein. PMID- 10364210 TI - The iron-oxygen reconstitution reaction in protein R2-Tyr-177 mutants of mouse ribonucleotide reductase. Epr and electron nuclear double resonance studies on a new transient tryptophan radical. AB - The ferrous iron/oxygen reconstitution reaction in protein R2 of mouse and Escherichia coli ribonucleotide reductase (RNR) leads to the formation of a stable protein-linked tyrosyl radical and a mu-oxo-bridged diferric iron center, both necessary for enzyme activity. We have studied the reconstitution reaction in three protein R2 mutants Y177W, Y177F, and Y177C of mouse RNR to investigate if other residues at the site of the radical forming Tyr-177 can harbor free radicals. In Y177W we observed for the first time the formation of a tryptophan radical in protein R2 of mouse RNR with a lifetime of several minutes at room temperature. We assign it to an oxidized neutral tryptophan radical on Trp-177, based on selective deuteration and EPR and electron nuclear double resonance spectroscopy in H2O and D2O solution. The reconstitution reaction at 22 degrees C in both Y177F and Y177C leads to the formation of a so-called intermediate X which has previously been assigned to an oxo (hydroxo)-bridged Fe(III)/Fe(IV) cluster. Surprisingly, in both mutants that do not have successor radicals as Trp. in Y177W, this cluster exists on a much longer time scale (several seconds) at room temperature than has been reported for X in E. coli Y122F or native mouse protein R2. All three mouse R2 mutants were enzymatically inactive, indicating that only a tyrosyl radical at position 177 has the capability to take part in the reduction of substrates. PMID- 10364211 TI - Molecular cloning and functional characterization of brefeldin A-ADP-ribosylated substrate. A novel protein involved in the maintenance of the Golgi structure. AB - Brefeldin A (BFA) is a fungal metabolite that disassembles the Golgi apparatus into tubular networks and causes the dissociation of coatomer proteins from Golgi membranes. We have previously shown that an additional effect of BFA is to stimulate the ADP-ribosylation of two cytosolic proteins of 38 and 50 kDa (brefeldin A-ADP-riboslyated substrate (BARS)) and that this effect greatly facilitates the Golgi-disassembling activity of the toxin. In this study, BARS has been purified from rat brain cytosol and microsequenced, and the BARS cDNA has been cloned. BARS shares high homology with two known proteins, C-terminal binding protein 1 (CtBP1) and CtBP2. It is therefore a third member of the CtBP family. The role of BARS in Golgi disassembly by BFA was verified in permeabilized cells. In the presence of dialyzed cytosol that had been previously depleted of BARS or treated with an anti-BARS antibody, BFA potently disassembled the Golgi. However, in cytosol complemented with purified BARS, or even in control cytosols containing physiological levels of BARS, the action of BFA on Golgi disassembly was strongly inhibited. These results suggest that BARS exerts a negative control on Golgi tubulation, with important consequences for the structure and function of the Golgi complex. PMID- 10364212 TI - Variation in the steady state kinetic parameters of wild type and mutant T5 5'-3' exonuclease with pH. Protonation of Lys-83 is critical for DNA binding. AB - T5 5'-3'-exonuclease is a member of a family of homologous 5'-nucleases essential for DNA replication and repair. We have measured the variation of the steady state parameters of the enzyme with pH. The log of the association constant of the enzyme and substrate is pH-independent between pH 5 and 7, but at higher pH, it decreases (gradient -0.91 +/- 0.1) with increasing pH. The log of the turnover number increases (gradient 0.9 +/- 0.01) with increasing pH until a pH independent plateau is reached. The T5 5'-3'-exonuclease-catalyzed reaction requires the protonation of a single residue for substrate binding, whereas kcat depends on a single deprotonation as demonstrated by the bell-shaped dependence of log (kcat/Km) on pH. To investigate the role of a conserved lysine (Lys-83), the pH profile of log (kcat/Km) of a K83A mutant was determined and found to increase with pH (gradient 1.01 +/- 0. 01) until a pH-independent plateau is reached. We therefore conclude that protonation of Lys-83 in the wild type protein facilitates DNA binding. The origin of the pH dependence of the kcat parameter of the wild type enzyme is discussed. PMID- 10364213 TI - Structural basis for the selectivity of the RGS protein, GAIP, for Galphai family members. Identification of a single amino acid determinant for selective interaction of Galphai subunits with GAIP. AB - GAIP is a regulator of G protein signaling (RGS) that accelerates the rate of GTP hydrolysis by some G protein alpha subunits. In the present studies, we have examined the structural basis for the ability of GAIP to discriminate among members of the Galphai family. Galphai1, Galphai3, and Galphao interacted strongly with GAIP, whereas Galphai2 interacted weakly and Galphas did not interact at all. A chimeric G protein composed of a Galphai2 N terminus and a Galphai1 C terminus interacted as strongly with GAIP as native Galphai1, whereas a chimeric N-terminal Galphai1 with a Galphai2 C terminus did not interact. These results suggest that the determinants responsible for GAIP selectivity between these two Galphais reside within the C-terminal GTPase domain of the G protein. To further localize residues contributing to G protein-GAIP selectivity, a panel of 15 site-directed Galphai1 and Galphai2 mutants were assayed. Of the Galphai1 mutants tested, only that containing a mutation at aspartate 229 located at the N terminus of Switch 3 did not interact with GAIP. Furthermore, the only Galphai2 variant that interacted strongly with GAIP contained a replacement of the corresponding Galphai2 Switch 3 residue (Ala230) with aspartate. To determine whether GAIP showed functional preferences for Galpha subunits that correlate with the binding data, the ability of GAIP to enhance the GTPase activity of purified alpha subunits was tested. GAIP catalyzed a 3-5-fold increase in the rate of GTP hydrolysis by Galphai1 and Galphai2(A230D) but no increase in the rate of Galphai2 and less than a 2-fold increase in the rate of Galphai1(D229A) under the same conditions. Thus, GAIP was able to discriminate between Galphai1 and Galphai2 in both binding and functional assays, and in both cases residue 229/230 played a critical role in selective recognition. PMID- 10364214 TI - Targeted expression of SV40 large T-antigen to visceral smooth muscle induces proliferation of contractile smooth muscle cells and results in megacolon. AB - Many pathological conditions result from the proliferation and de-differentiation of smooth muscle cells leading to impaired contractility of the muscle. Here we show that targeted expression of SV40 large T-antigen to visceral smooth muscle cells in vivo results in increased smooth muscle cell proliferation without de differentiation or decreased contractility. These data suggest that the de differentiation and proliferation of smooth muscle cells, seen in many pathological states, may be independently regulated. In the T-antigen transgenic mice the increased smooth muscle cell proliferation results in thickening of the distal colon. Consequently the distal colon becomes hyper-contractile and impedes the flow of digesta through the colon resulting in enlargement of the colon oral to the obstruction. These transgenic mice thus represent a novel model of megacolon that results from increased smooth muscle cell proliferation rather than altered neuronal innervation. PMID- 10364215 TI - Antichymotrypsin interaction with chymotrypsin. Intermediates on the way to inhibited complex formation. AB - Serpins form enzymatically inactive covalent complexes (designated E*I*) with their target proteinases, corresponding most likely to the acyl enzyme that resembles the normal intermediate in substrate turnover. Formation of E*I* involves large changes in the conformation of the reactive center loop (residues P17 to P9') and of the serpin molecule in general. The "hinge" region of the reactive center loop, including residues P10-P14, shows facile movement in and out of beta-sheet A, and this movement appears to be crucial in determining whether E*I* is formed (the inhibitor pathway) or whether I is rapidly hydrolyzed to I* (the substrate pathway). Here, we report stopped-flow and rapid quench studies investigating the pH dependence of the conversion of the alpha1 antichymotrypsin.alpha-chymotrypsin encounter complex, E.I, to E*I*. These studies utilize fluorescent derivatives of cysteine variants of alpha1 antichymotrypsin at the P11 and P13 residues. Our results demonstrate three identifiable intermediates, EIa, EIb, and EIc, between E.I and E*I* and permit informed speculation regarding the nature of these intermediates. Partitioning between inhibitor and substrate pathways occurs late in the process of E*I* formation, most likely from a species occurring between EIc and E*I*. PMID- 10364216 TI - Aldolase mediates the association of F-actin with the insulin-responsive glucose transporter GLUT4. AB - To identify potential proteins interacting with the insulin-responsive glucose transporter (GLUT4), we generated fusion proteins of glutathione S-transferase (GST) and the final 30 amino acids from GLUT4 (GST-G4) or GLUT1 (GST-G1). Incubation of these carboxyl-terminal fusion proteins with adipocyte cell extracts revealed a specific interaction of GLUT4 with fructose 1, 6-bisphosphate aldolase. In the presence of aldolase, GST-G4 but not GST-G1 was able to co pellet with filamentous (F)-actin. This interaction was prevented by incubation with the aldolase substrates, fructose 1,6-bisphosphate or glyceraldehyde 3 phosphate. Immunofluorescence confocal microscopy demonstrated a significant co localization of aldolase and GLUT4 in intact 3T3L1 adipocytes, which decreased following insulin stimulation. Introduction into permeabilized 3T3L1 adipocytes of fructose 1,6-bisphosphate or the metabolic inhibitor 2-deoxyglucose, two agents that disrupt the interaction between aldolase and actin, inhibited insulin stimulated GLUT4 exocytosis without affecting GLUT4 endocytosis. Furthermore, microinjection of an aldolase-specific antibody also inhibited insulin-stimulated GLUT4 translocation. These data suggest that aldolase functions as a scaffolding protein for GLUT4 and that glucose metabolism may provide a negative feedback signal for the regulation of glucose transport by insulin. PMID- 10364217 TI - Circadian regulation of cAMP response element-mediated gene expression in the suprachiasmatic nuclei. AB - A program of stringently-regulated gene expression is thought to be a fundamental component of the circadian clock. Although recent work has implicated a role for E-box-dependent transcription in circadian rhythmicity, the contribution of other enhancer elements has yet to be assessed. Here, we report that cells of the suprachiasmatic nuclei (SCN) exhibit a prominent circadian oscillation in cAMP response element (CRE)-mediated gene expression. Maximal reporter gene expression occurred from late-subjective night to mid-subjective day. Cycling of CRE dependent transcription was not observed in other brain regions, including the supraoptic nucleus and piriform cortex. Levels of the phospho-active form of the transcription factor CREB (P-CREB) varied as a function of circadian time. Peak P CREB levels occurred during the mid- to late-subjective night. Furthermore, photic stimulation during the subjective night, but not during the subjective day, triggered a marked increase in CRE-mediated gene expression in the SCN. Reporter gene experiments showed that activation of the p44/42 mitogen-activated protein kinase signaling cascade is required for Ca2+-dependent stimulation of CRE-mediated transcription in the SCN. These findings reveal the CREB/CRE transcriptional pathway to be circadian-regulated within the SCN, and raise the possibility that this pathway provides signaling information essential for normal clock function. PMID- 10364219 TI - Effector recognition by the small GTP-binding proteins Ras and Ral. AB - The Ral effector protein RLIP76 (also called RIP/RalBP1) binds to Ral.GTP via a region that shares no sequence homology with the Ras-binding domains of the Ser/Thr kinase c-Raf-1 and the Ral-specific guanine nucleotide exchange factors. Whereas the Ras-binding domains have a similar ubiquitin-like structure, the Ral binding domain of RLIP was predicted to comprise a coiled-coil region. In order to obtain more information about the specificity and the structural mode of the interaction between Ral and RLIP, we have performed a sequence space and a mutational analysis. The sequence space analysis of a comprehensive nonredundant assembly of Ras-like proteins strongly indicated that positions 36 and 37 in the core of the effector region are tree-determinant positions for all subfamilies of Ras-like proteins and dictate the specificity of the interaction of these GTPases with their effector proteins. Indeed, we could convert the specific interaction with Ras effectors and RLIP by mutating these residues in Ras and Ral. We therefore conclude that positions 36 and 37 are critical for the discrimination between Ras and Ral effectors and that, despite the absence of sequence homology between the Ral-binding and the Ras-binding domains, their mode of interaction is most probably similar. PMID- 10364218 TI - Morphological changes and detachment of adherent cells induced by p122, a GTPase activating protein for Rho. AB - We recently cloned a novel signaling molecule, p122, that shows a GTPase activating activity specific for Rho and the ability to enhance the phosphatidylinositol 4,5-bisphosphate-hydrolyzing activity of phospholipase C delta1 in vitro. Here we analyzed the in vivo function of p122. Microinjection of the GTPase-activating domain of p122 suppressed the formation of stress fibers and focal adhesions induced by lysophosphatidic acid, suggesting a GTPase activating activity for Rho as in in vitro. Transfection of p122 also induced the disassembly of stress fibers and the morphological rounding of various adherent cells. Analyses using deletion and point mutants demonstrated that the GTPase activating domain of p122 is responsible for the morphological changes and detachment and that arginine residues at positions 668 and 710 and a lysine residue at position 706 in the GTPase-activating domain are essential. Using Fluo 3-based Ca2+ microscopy, we found that p122 evoked a rapid elevation of intracellular Ca2+ levels, suggesting that p122 stimulates the phosphatidylinositol 4, 5-bisphosphate-hydrolyzing activity of phospholipase C delta1. These results demonstrate that p122 synergistically functions as a GTPase activating protein specific for Rho and an activator of phospholipase C delta1 in vivo and induces morphological changes and detachment through cytoskeletal reorganization. PMID- 10364220 TI - Structure and function of human prepro-orexin gene. AB - Orexin-A and -B are recently identified potent orexigenic peptides that are derived from the same precursor peptide and are highly specifically localized in neurons located in the lateral hypothalamic area, a region classically implicated in feeding behavior. We cloned the whole length of the human prepro-orexin gene and corresponding cDNA. The human prepro-orexin mRNA was predicted to encode a 131-residue precursor peptide (prepro-orexin). The human prepro-orexin gene consists of two exons and one intron distributed over 1432 base pairs. The 143 base pair first exon includes the 5'-untranslated region and a small part of the coding region that encodes the first seven residues of the secretory signal sequence. The second exon contains the remaining portion of the open reading frame and 3'-untranslated region. The 3.2 kilobase pairs of the 5'-upstream region from a cloned human prepro-orexin gene promoter is sufficient to direct the expression of the Escherichia coli beta-galactosidase (lacZ) gene in transgenic mice to neurons in the lateral hypothalamic area and adjacent regions. The lacZ-positive neurons were positively stained with anti-orexin antibody but not with anti-melanin-concentrating hormone antibody. These findings suggest that this genomic fragment contains all the necessary elements for appropriate expression of the gene and will be useful for the targeted expression of the exogenous gene in orexin-containing neurons. These mice might also be useful for examining the molecular mechanisms by which orexin gene expression is regulated. PMID- 10364221 TI - Molecular cloning, enzymatic characterization, developmental expression, and cellular localization of a mouse cytochrome P450 highly expressed in kidney. AB - A cDNA encoding a new cytochrome P450 was isolated from a mouse liver library. Sequence analysis reveals that this 1,886-base pair cDNA encodes a 501-amino acid polypeptide that is 69-74% identical to CYP2J subfamily P450s and is designated CYP2J5. Recombinant CYP2J5 was co-expressed with NADPH-cytochrome P450 oxidoreductase in Sf9 cells using a baculovirus system. Microsomal fractions of CYP2J5/NADPH-cytochrome P450 oxidoreductase-transfected cells metabolize arachidonic acid to 14,15-, 11,12-, and 8, 9-epoxyeicosatrienoic acids and 11- and 15-hydroxyeicosatetraenoic acids (catalytic turnover, 4.5 nmol of product/nmol of cytochrome P450/min at 37 degrees C); thus CYP2J5 is enzymologically distinct. Northern analysis reveals that CYP2J5 transcripts are most abundant in mouse kidney and present at lower levels in liver. Immunoblotting using a polyclonal antibody against a CYP2J5-specific peptide detects a protein with the same electrophoretic mobility as recombinant CYP2J5 most abundantly in mouse kidney microsomes. CYP2J5 is regulated during development in a tissue-specific fashion. In the kidney, CYP2J5 is present before birth and reaches maximal levels at 2-4 weeks of age. In the liver, CYP2J5 is absent prenatally and during the early postnatal period, first appears at 1 week, and then remains relatively constant. Immunohistochemical staining of kidney sections with anti-human CYP2J2 IgG reveals that CYP2J protein(s) are present primarily in the proximal tubules and collecting ducts, sites where the epoxyeicosatrienoic acids are known to modulate fluid/electrolyte transport and mediate hormonal action. In situ hybridization confirms abundant CYP2J5 mRNA within tubules of the renal cortex and outer medulla. Epoxyeicosatrienoic acids are endogenous constituents of mouse kidney thus providing direct evidence for the in vivo metabolism of arachidonic acid by the mouse renal epoxygenase(s). Based on these data, we conclude that CYP2J5 is an enzymologically distinct, developmentally regulated, protein that is localized to specific nephron segments and contributes to the oxidation of endogenous renal arachidonic acid pools. In light of the well documented effects of epoxyeicosatrienoic acids in modulating renal tubular transport processes, we postulate that CYP2J5 products play important functional roles in the kidney. PMID- 10364222 TI - The mechanism of adenosine formation in cells. Cloning of cytosolic 5' nucleotidase-I. AB - Adenosine increases blood flow and decreases excitatory nerve firing. In the heart, it reduces rate and force of contraction and preconditions the heart against injury by prolonged ischemia. Based on indirect kinetic arguments, an AMP selective cytosolic 5'-nucleotidase designated cN-I has been implicated in adenosine formation during ATP breakdown. The molecular identity of cN-I is unknown, although an IMP/GMP-selective cytosolic 5'-nucleotidase (cN-II) and an ecto-5'-nucleotidase (e-N) have been cloned. We utilized the high abundance of cN I in pigeon heart to purify a 40-kDa subunit for partial protein sequencing and subsequent cDNA cloning. We obtained a full-length clone encoding a novel 40-kDa peptide, unrelated to cN-II or e-N, that was most abundant in heart, brain, and breast muscle. Immunolocalization in heart showed a striated cytoplasmic location, suggesting association with contractile elements. Transient expression in COS-7 cells, generated a 5'-nucleotidase that catalyzed adenosine formation from AMP, which was increased during ATP catabolism. In conclusion, the cloning and expression of cN-I provides definitive evidence of its ability to produce adenosine during ATP breakdown. PMID- 10364223 TI - Coupled inositide phosphorylation and phospholipase D activation initiates clathrin-coat assembly on lysosomes. AB - Adaptors appear to control clathrin-coat assembly by determining the site of lattice polymerization but the nucleating events that target soluble adaptors to an appropriate membrane are poorly understood. Using an in vitro model system that allows AP-2-containing clathrin coats to assemble on lysosomes, we show that adaptor recruitment and coat initiation requires phosphatidylinositol 4,5 bisphosphate (PtdIns(4,5)P2) synthesis. PtdIns(4,5)P2 is generated on lysosomes by the sequential action of a lysosome-associated type II phosphatidylinositol 4 kinase and a soluble type I phosphatidylinositol 4-phosphate 5-kinase. Phosphatidic acid, which potently stimulates type I phosphatidylinositol 4 phosphate 5-kinase activity, is generated on the bilayer by a phospholipase D1 like enzyme located on the lysosomal surface. Quenching phosphatidic acid function with primary alcohols prevents the synthesis of PtdIns(4, 5)P2 and blocks coat assembly. Generating phosphatidic acid directly on lysosomes with exogenous bacterial phospholipase D in the absence of ATP still drives adaptor recruitment and limited coat assembly, indicating that PtdIns(4,5)P2 functions, at least in part, to activate the PtdIns(4,5)P2-dependent phospholipase D1. These results provide the first direct evidence for the involvement of anionic phospholipids in clathrin-coat assembly on membranes and define the enzymes responsible for the production of these important lipid mediators. PMID- 10364224 TI - Identification of the cell cycle regulator VCP (p97/CDC48) as a substrate of the band 4.1-related protein-tyrosine phosphatase PTPH1. AB - The human band 4.1-related protein-tyrosine phosphatase PTPH1 was introduced into NIH3T3 cells under the control of a tetracycline-repressible promoter. Ectopic expression of wild type PTPH1 dramatically inhibited cell growth, whereas a catalytically impaired mutant showed no effect. To identify the direct target of PTPH1 in the cell, we generated a substrate-trapping mutant, in which an invariant aspartate residue was changed to alanine (D811A in PTPH1). The PTPH1 D811A mutant trapped primarily a 97-kDa tyrosine-phosphorylated protein, which was determined to be VCP (also named p97 or yeast CDC48), from various cell lysates in vitro. However, when expressed in mammalian cells, the D811A mutant was observed to contain high levels of phosphotyrosine and did not trap substrates. Mutation of tyrosine 676 to phenylalanine (Y676F) in the PTPH1-D811A mutant led to a marked reduction in phosphotyrosine content. Furthermore, this double mutant specifically trapped VCP in vivo and recognized the C-terminal tyrosines of VCP, whose phosphorylation is important for cell cycle progression in yeast. Like wild type PTPH1, this double mutant also inhibited cell proliferation. Moreover, induction of wild type PTPH1 resulted in specific dephosphorylation of VCP without changing the overall phosphotyrosine profile of the cells. VCP has been implicated in control of a variety of membrane functions, including membrane fusions, and is a regulator of the cell cycle. Our results suggest that PTPH1 may exert its effects on cell growth through dephosphorylation of VCP, thus implicating tyrosine phosphorylation as an important regulator of VCP function. PMID- 10364226 TI - An antagonist of cADP-ribose inhibits arrhythmogenic oscillations of intracellular Ca2+ in heart cells. AB - Oscillations of Ca2+ in heart cells are a major underlying cause of important cardiac arrhythmias, and it is known that Ca2+-induced release of Ca2+ from intracellular stores (the sarcoplasmic reticulum) is fundamental to the generation of such oscillations. There is now evidence that cADP-ribose may be an endogenous regulator of the Ca2+ release channel of the sarcoplasmic reticulum (the ryanodine receptor), raising the possibility that cADP-ribose may influence arrhythmogenic mechanisms in the heart. 8-Amino-cADP-ribose, an antagonist of cADP-ribose, suppressed oscillatory activity associated with overloading of intracellular Ca2+ stores in cardiac myocytes exposed to high doses of the beta adrenoreceptor agonist isoproterenol or the Na+/K+-ATPase inhibitor ouabain. The oscillations suppressed by 8-amino-cADP-ribose included intracellular Ca2+ waves, spontaneous action potentials, after-depolarizations, and transient inward currents. Another antagonist of cADP-ribose, 8-bromo-cADP-ribose, was also effective in suppressing isoproterenol-induced oscillatory activity. Furthermore, in the presence of ouabain under conditions in which there was no arrhythmogenesis, exogenous cADP-ribose was found to be capable of triggering spontaneous contractile and electrical activity. Because enzymatic machinery for regulating the cytosolic cADP-ribose concentration is present within the cell, we propose that 8-amino-cADP-ribose and 8-bromo-cADP-ribose suppress cytosolic Ca2+ oscillations by antagonism of endogenous cADP-ribose, which sensitizes the Ca2+ release channels of the sarcoplasmic reticulum to Ca2+. PMID- 10364225 TI - Mouse ATF-2 null mutants display features of a severe type of meconium aspiration syndrome. AB - Mouse null mutants of transcription factor ATF-2 were generated by the gene targeting method. They died shortly after birth and displayed symptoms of severe respiratory distress with lungs filled with meconium. These features are similar to those of a severe type of human meconium aspiration syndrome. The increased expression of the hypoxia inducible genes suggests that hypoxia occurs in the mutant embryos and that it may lead to strong gasping respiration with consequent aspiration of the amniotic fluid containing meconium. A reduced number of cytotrophoblast cells in the mutant placenta was found and may be responsible for an insufficient supply of oxygen prior to birth. Using the cDNA subtraction and microarray-based expression monitoring method, the expression level of the platelet-derived growth factor receptor alpha gene, which plays an important role in the proliferation of trophoblasts, was found to be low in the cytotrophoblasts of the mutant placenta. In addition, ATF-2 can trans-activate the PDGF receptor alpha gene promoter in the co-transfection assay. These results indicate the important role of ATF-2 in the formation of the placenta and the relationship between placental anomalies and neonatal respiratory distress. The ATF-2 null mutants should enhance our understanding of the mechanism of severe neonatal respiratory distress. PMID- 10364227 TI - Multiple p44 genes encoding major outer membrane proteins are expressed in the human granulocytic ehrlichiosis agent. AB - Human granulocytic ehrlichiosis (HGE) is caused by infection with an obligatory intracellular bacterium, the HGE agent. We previously cloned a gene encoding HGE agent 44-kDa major outer membrane protein and designated it p44. In this study, we (i) identified five different mRNAs that are transcribed from p44-homologous genes in the HGE agent cultivated in HL-60 cells; (ii) cloned genes corresponding to the mRNAs from the genomic DNA of the HGE agent; (iii) showed that the genes being expressed were not clustered in the HGE agent genome; (iv) estimated that a minimum copy number of the p44-homologous genes in the genome is 18; (v) detected two different P44-homologous proteins expressed by the HGE agent; and (vi) demonstrated existence of antibodies specific to the two proteins in sera from patients with HGE. These findings showed that p44 multigenes have several active expression sites and the expression is regulated at transcriptional level, suggesting a potentially unique mechanism for generating the diversity in major antigenic outer membrane proteins of the HGE agent. Characterization of p44 homologous genes expressed by the HGE agent in a tissue culture would assist in understanding a role of the p44 multigene family in pathogenesis and immune response in HGE. PMID- 10364228 TI - Identification of the stef gene that encodes a novel guanine nucleotide exchange factor specific for Rac1. AB - The Rho family GTPases are involved in a variety of cellular events by changing the organization of actin cytoskeletal networks in response to extracellular signals. However, it is not clearly known how their activities are spatially and temporally regulated. Here we report the identification of a novel guanine nucleotide exchange factor for Rac1, STEF, which is related in overall amino acid sequence and modular structure to mouse Tiam1 and Drosophila SIF proteins. STEF protein contains two pleckstrin homology domains, a PDZ domain and a Dbl homology domain. The in vitro assay showed that STEF protein specifically enhanced the dissociation of GDP from Rac1 but not that from either RhoA or Cdc42. Expression of a truncated STEF protein in culture cells induced membrane ruffling with altered actin localization, which implies that this protein also activates Rac1 in vivo. The stef transcript was observed in restricted parts of mice, including cartilaginous tissues and the cortical plate of the central nervous system during embryogenesis. These findings suggested that STEF protein participates in the control of cellular events in several developing tissues, possibly changing the actin cytoskeletal network by activating Rac1. PMID- 10364229 TI - Crystal structure determinations of oxidized and reduced pseudoazurins from Achromobacter cycloclastes. Concerted movement of copper site in redox forms with the rearrangement of hydrogen bond at a remote histidine. AB - The crystal structures of oxidized and reduced pseudoazurins from a denitrifying bacterium, Achromobacter cycloclastes IAM1013, have been determined at 1.35- and 1.6-A resolutions, respectively. The copper site in the oxidized state exhibits a distorted tetrahedral structure like those of other pseudoazurins. However, not only a small change of the copper geometry, but concerted peptide bond flips are identified. The imidazole ring of remote His6 has a hydrogen bonding distance of 2.73 A between N-delta1(His6) and O-gamma1(Thr36) in the oxidized protein. When the protein is reduced at pH 6.0, the imidazole ring rotates by 30.3 degrees and moves 1.00 A away from the position of the oxidized state. A new hydrogen bond between N-epsilon2(His6) and O-epsilon1(Glu4) is formed with a distance of 3.03 A, while the hydrogen bond between N-delta1(His6)-O-gamma1(Thr36) is maintained with an interatomic distance of 2.81 A. A concomitant peptide bond flip of main chain between Ile34 and Thr36 occurs. PMID- 10364230 TI - Ligand exchange during unfolding of cytochrome c. AB - The productive folding pathway of cytochrome c passes through an obligatory HW intermediate in which the heme is coordinated by a solvent water molecule and a native ligand, His-18, prior to the formation of the folded HM state with both the native His-18 and Met-80 heme coordination. Two off pathway intermediates, a five-coordinated state (5C) and a bis-histidine state (HH), were also identified during the folding reaction. In the present work, the thermodynamics and the kinetics of the unfolding reaction of cytochrome c were investigated with resonance Raman scattering, tryptophan fluorescence spectroscopy, and circular dichroism. The objective of these experiments was to determine if the protein opens up and diverges into the differing heme ligation states through a many pathway mechanism or if it passes through intermediate states analogous to those observed during the folding reaction. Equilibrium unfolding results indicate that, in contrast to 5C, the stability of HH with respect to HW decreases as the concentration of GdnHCl increases. The difference in their response to the denaturant indicates that the polypeptide structure of 5C is relatively loose as compared with HH in which the polypeptide is misfolded. Time-resolved resonance Raman measurements show that strikingly similar ligand exchange reactions occur during unfolding as were observed during folding. Combined with fluorescence data, a kinetic model is proposed in which local structural rearrangements controlled by heme ligand exchange reactions appear prior to the global relaxation of the polypeptide chain. PMID- 10364232 TI - Binding of nucleotide triphosphates to cardiotoxin analogue II from the Taiwan cobra venom (Naja naja atra). Elucidation of the structural interactions in the dATP-cardiotoxin analogue ii complex. AB - Snake venom cardiotoxins have been recently shown to block the enzymatic activity of phospholipid protein kinase and Na+,K+-ATPase. To understand the molecular basis for the inhibitory effects of cardiotoxin on the action of these enzymes, the nucleotide triphosphate binding ability of cardiotoxin analogue II (CTX II) from the Taiwan cobra (Naja naja atra) venom is investigated using a variety of spectroscopic techniques such as fluorescence, circular dichroism, and two dimensional NMR. CTX II is found to bind to all the four nucleotide triphosphates (ATP, UTP, GTP, and CTP) with similar affinity. Detailed studies of the binding of dATP to CTX II indicated that the toxin molecule is significantly stabilized in the presence of the nucleotide. Molecular modeling, based on the NOEs observed for the dATP.CTX II complex, reveals that dATP binds to the CTX II molecule at the groove enclosed between the N- and C-terminal ends of the toxin molecule. Based on the results obtained in the present study, a molecular mechanism to account for the inhibition of the enzymatic activity of the phospholipid sensitive protein kinase and Na+,K+-ATPase is also proposed. PMID- 10364231 TI - PARP-2, A novel mammalian DNA damage-dependent poly(ADP-ribose) polymerase. AB - Poly(ADP-ribosylation) is a post-translational modification of nuclear proteins in response to DNA damage that activates the base excision repair machinery. Poly(ADP-ribose) polymerase which we will now call PARP-1, has been the only known enzyme of this type for over 30 years. Here, we describe a cDNA encoding a 62-kDa protein that shares considerable homology with the catalytic domain of PARP-1 and also contains a basic DNA-binding domain. We propose to call this enzyme poly(ADP-ribose) polymerase 2 (PARP-2). The PARP-2 gene maps to chromosome 14C1 and 14q11.2 in mouse and human, respectively. Purified recombinant mouse PARP-2 is a damaged DNA-binding protein in vitro and catalyzes the formation of poly(ADP-ribose) polymers in a DNA-dependent manner. PARP-2 displays automodification properties similar to PARP-1. The protein is localized in the nucleus in vivo and may account for the residual poly(ADP-ribose) synthesis observed in PARP-1-deficient cells, treated with alkylating agents or hydrogen peroxide. PMID- 10364233 TI - Identification of two amino acids within the EIIIA (ED-A) segment of fibronectin constituting the epitope for two function-blocking monoclonal antibodies. AB - Alternative splicing of the fibronectin gene transcript gives rise to a group of adhesive glycoproteins showing restricted spatial and temporal expression during embryonic development, tumor growth, and tissue repair. Alternative splicing occurs in three segments termed EIIIB, EIIIA, and V. The EIIIA (or ED-A) segment of fibronectin is expressed prominently but transiently in healing wounds coincident with fibroblast expression of an activation marker, smooth muscle cell alpha-actin. A monoclonal antibody (IST-9) to the EIIIA segment blocks transforming growth factor-beta-mediated smooth muscle cell alpha-actin expression by fibroblasts in culture. A second monoclonal antibody (DH1) blocks chondrocyte condensation in chicken embryos. We find that IST-9 and DH1 react with human, rat, and chicken but not with mouse or frog EIIIA, suggesting that His44 may be important for antibody binding. A series of deletion mutants of rat EIIIA, constructed as glutathione S-transferase fusion proteins, do not react with either IST-9, DH1, or a third monoclonal antibody (3E2). Mutations of pairs of amino acids to alanine have little effect, except for either (Val34Thr35) or (Tyr36Ser37), which are located in a beta strand upstream from His44. For these double mutants, the binding to all three monoclonal antibodies is markedly reduced. By contrast, single mutants at Thr35, Tyr36, or Ser37 retain full activity, suggesting that the epitope for these antibodies is determined in part by conformation. Alanine-scanning mutagenesis of rat EIIIA demonstrates the importance of Ile43 and His44 for binding. Mutation of frog EIIIA (normally Val43Lys44) to rat (Ile43His44) is sufficient to restore fully IST-9 binding and much of the activity of DH1 and 3E2. Our findings demonstrate that the function blocking antibodies, IST-9 and DH1, bind to the Ile43 and His44 residues in a conformationally dependent fashion, implicating the loop region encompassing both residues as critical for mediating EIIIA function. PMID- 10364234 TI - Mammalian homologues of the Drosophila slit protein are ligands of the heparan sulfate proteoglycan glypican-1 in brain. AB - Using an affinity matrix in which a recombinant glypican-Fc fusion protein expressed in 293 cells was coupled to protein A-Sepharose, we have isolated from rat brain at least two proteins that were detected by SDS-polyacrylamide gel electrophoresis as a single 200-kDa silver-stained band, from which 16 partial peptide sequences were obtained by nano-electrospray tandem mass spectrometry. Mouse expressed sequence tags containing two of these peptides were employed for oligonucleotide design and synthesis of probes by polymerase chain reaction and enabled us to isolate from a rat brain cDNA library a 4.1-kilobase clone that encoded two of our peptide sequences and represented the N-terminal portion of a protein containing a signal peptide and three leucine-rich repeats. Comparisons with recently published sequences also showed that our peptides were derived from proteins that are members of the Slit/MEGF protein family, which share a number of structural features such as N-terminal leucine-rich repeats and C-terminal epidermal growth factor-like motifs, and in Drosophila Slit is necessary for the development of midline glia and commissural axon pathways. All of the five known rat and human Slit proteins contain 1523-1534 amino acids, and our peptide sequences correspond best to those present in human Slit-1 and Slit-2. Binding of these ligands to the glypican-Fc fusion protein requires the presence of the heparan sulfate chains, but the interaction appears to be relatively specific for glypican-1 insofar as no other identified heparin-binding proteins were isolated using our affinity matrix. Northern analysis demonstrated the presence of two mRNA species of 8. 6 and 7.5 kilobase pairs using probes based on both N- and C terminal sequences, and in situ hybridization histochemistry showed that these glypican-1 ligands are synthesized by neurons, such as hippocampal pyramidal cells and cerebellar granule cells, where we have previously also demonstrated glypican-1 mRNA and immunoreactivity. Our results therefore indicate that Slit family proteins are functional ligands of glypican-1 in nervous tissue and suggest that their interactions may be critical for certain stages of central nervous system histogenesis. PMID- 10364235 TI - Human exonuclease 1 functionally complements its yeast homologues in DNA recombination, RNA primer removal, and mutation avoidance. AB - Yeast exonuclease 1 (Exo1) is induced during meiosis and plays an important role in DNA homologous recombination and mismatch correction pathways. The human homolog, an 803-amino acid protein, shares 55% similarity to the yeast Exo1. In this report, we show that the enzyme functionally complements Saccharomyces cerevisiae Exo1 in recombination of direct repeat DNA fragments, UV resistance, and mutation avoidance by in vivo assays. Furthermore, the human enzyme suppresses the conditional lethality of a rad27Delta mutant, symptomatic of defective RNA primer removal. The purified recombinant enzyme not only displays 5'-3' double strand DNA exonuclease activity, but also shows an RNase H activity. This result indicates a back-up function of exonuclease 1 to flap endonuclease-1 in RNA primer removal during lagging strand DNA synthesis. PMID- 10364236 TI - Differential involvement of Galpha12 and Galpha13 in receptor-mediated stress fiber formation. AB - The ubiquitously expressed heterotrimeric guanine nucleotide-binding proteins (G proteins) G12 and G13 have been shown to activate the small GTPase Rho. Rho stimulation leads to a rapid remodeling of the actin cytoskeleton and subsequent stress fiber formation. We investigated the involvement of G12 or G13 in stress fiber formation induced through a variety of Gq/G11-coupled receptors. Using fibroblast cell lines derived from wild-type and Galphaq/Galpha11-deficient mice, we show that agonist-dependent activation of the endogenous receptors for thrombin or lysophosphatidic acid and of the heterologously expressed bradykinin B2, vasopressin V1A, endothelin ETA, and serotonin 5-HT2C receptors induced stress fiber formation in either the presence or absence of Galphaq/Galpha11. Stress fiber assembly induced through the muscarinic M1 and the metabotropic glutamate subtype 1alpha receptors was dependent on Gq/G11 proteins. The activation of the Gq/G11-coupled endothelin ETB and angiotensin AT1A receptors failed to induce stress fiber formation. Lysophosphatidic acid, B2, and 5-HT2C receptor-mediated stress fiber formation was dependent on Galpha13 and involved epidermal growth factor (EGF) receptors, whereas thrombin, ETA, and V1A receptors induced stress fiber accumulation via Galpha12 in an EGF receptor-independent manner. Our data demonstrate that many Gq/G11-coupled receptors induce stress fiber assembly in the absence of Galphaq and Galpha11 and that this involves either a Galpha12 or a Galpha13/EGF receptor-mediated pathway. PMID- 10364237 TI - Ataxia telangiectasia-mutated gene product inhibits DNA damage-induced apoptosis via ceramide synthase. AB - DNA double-stranded breaks (dsb) activate surveillance systems that identify DNA damage and either initiate repair or signal cell death. Failure of cells to undergo appropriate death in response to DNA damage leads to misrepair, mutations, and neoplastic transformation. Pathways linking DNA dsb to reproductive or apoptotic death are virtually unknown. Here we report that metabolic incorporation of 125I-labeled 5-iodo-2'deoxyuridine, which produces DNA dsb, signaled de novo ceramide synthesis by post-translational activation of ceramide synthase (CS) and apoptosis. CS activation was obligatory, since fumonisin B1, a fungal pathogen that acts as a specific CS inhibitor, abrogated DNA damage-induced death. X-irradiation yielded similar results. Furthermore, inhibition of apoptosis using the peptide caspase inhibitor benzyloxycarbonyl-Val Ala-Asp fluoromethylketone did not affect CS activation, indicating this event is not a consequence of induction of apoptosis. ATM, the gene mutated in ataxia telangiectasia, is a member of the phosphatidylinositol 3-kinase family that constitutes the DNA damage surveillance/repair system. Epstein-Barr virus immortalized B cell lines from six ataxia telangiectasia patients with different mutations exhibited radiation-induced CS activation, ceramide generation, and apoptosis, whereas three lines from normal patients failed to manifest these responses. Stable transfection of wild type ATM cDNA reversed these events, whereas antisense inactivation of ataxia telangiectasia-mutated gene product in normal B cells conferred the ataxia telangiectasia phenotype. We propose that one of the functions of ataxia telangiectasia-mutated gene product is to constrain activation of CS, thereby regulating DNA damage-induced apoptosis. PMID- 10364238 TI - A critical DnaA box directs the cooperative binding of the Escherichia coli DnaA protein to the plasmid RK2 replication origin. AB - The requirement of DnaA protein binding for plasmid RK2 replication initiation the Escherichia coli was investigated by constructing mutations in the plasmid replication origin that scrambled or deleted each of the four upstream DnaA boxes. Altered origins were analyzed for replication activity in vivo and in vitro and for binding to the E. coli DnaA protein using a gel mobility shift assay and DNase I footprinting. Most strikingly, a mutation in one of the boxes, box 4, abolished replication activity and eliminated stable DnaA protein binding to all four boxes. Unlike DnaA binding to the E. coli origin, oriC, DnaA binding to two of the boxes (boxes 4 and 3) in the RK2 origin, oriV, is cooperative with box 4 acting as the "organizer" for the formation of the DnaA-oriV nucleoprotein complex. Interestingly, the inversion of box 4 also abolished replication activity, but did not result in a loss of binding to the other boxes. However, DnaA binding to this mutant origin was no longer cooperative. These results demonstrate that the sequence, position, and orientation of box 4 are crucial for cooperative DnaA binding and the formation of a nucleoprotein structure that is functional for the initiation of replication. PMID- 10364239 TI - The plasmid ColIb-P9 antisense Inc RNA controls expression of the RepZ replication protein and its positive regulator repY with different mechanisms. AB - The autonomous replication region of plasmid ColIb-P9 contains repZ encoding the RepZ replication protein, and inc and repY as the negative and positive regulators of repZ translation, respectively. inc encodes the antisense Inc RNA, and repY is a short open reading frame upstream of repZ. Translation of repY enables repZ translation by inducing formation of a pseudoknot containing stem loop I, which base pairs with the sequence preceding the repZ start codon. Inc RNA inhibits both repY translation and formation of the pseudoknot by binding to the loop I. To investigate control of repY expression by Inc RNA, we isolated a number of mutations that express repY in the presence of Inc RNA. One class of mutations delete a part of another stem-loop (II), which derepresses repY expression by initiating translation at codon 10 (GUG), located within this structure. Point mutations in stem-loop II can also derepress repY translation, and the introduction of compensatory base-changes restores control of repY translation. These results not only indicate that suppressing a cryptic start codon by secondary structure is important for maintaining the translational control of repZ but also demonstrate that the position of start site for repY translation is critical for its control by Inc RNA. Thus, Inc RNA controls repY translation by binding in the vicinity of the start codon, in contrast to the control of repZ expression at the level of loop-loop interaction. PMID- 10364240 TI - The role of glycogen synthase kinase 3beta in insulin-stimulated glucose metabolism. AB - To characterize the contribution of glycogen synthase kinase 3beta (GSK3beta) inactivation to insulin-stimulated glucose metabolism, wild-type (WT-GSK), catalytically inactive (KM-GSK), and uninhibitable (S9A-GSK) forms of GSK3beta were expressed in insulin-responsive 3T3-L1 adipocytes using adenovirus technology. WT-GSK, but not KM-GSK, reduced basal and insulin-stimulated glycogen synthase activity without affecting the -fold stimulation of the enzyme by insulin. S9A-GSK similarly decreased cellular glycogen synthase activity, but also partially blocked insulin stimulation of the enzyme. S9A-GSK expression also markedly inhibited insulin stimulation of IRS-1-associated phosphatidylinositol 3 kinase activity, but only weakly inhibited insulin-stimulated Akt/PKB phosphorylation and glucose uptake, with no effect on GLUT4 translocation. To further evaluate the role of GSK3beta in insulin signaling, the GSK3beta inhibitor lithium was used to mimic the consequences of insulin-stimulated GSK3beta inactivation. Although lithium stimulated the incorporation of glucose into glycogen and glycogen synthase enzyme activity, the inhibitor was without effect on GLUT4 translocation and pp70 S6 kinase. Lithium stimulation of glycogen synthesis was insensitive to wortmannin, which is consistent with its acting directly on GSK3beta downstream of phosphatidylinositol 3-kinase. These data support the hypothesis that GSK3beta contributes to insulin regulation of glycogen synthesis, but is not responsible for the increase in glucose transport. PMID- 10364241 TI - Cytochrome c and dATP-mediated oligomerization of Apaf-1 is a prerequisite for procaspase-9 activation. AB - To elucidate the mechanism of activation of procaspase-9 by Apaf-1, we produced recombinant full-length Apaf-1 and purified it to complete homogeneity. Here we show using gel filtration that full-length Apaf-1 exists as a monomer that can be transformed to an oligomeric complex made of at least eight subunits after binding to cytochrome c and dATP. Apaf-1 binds to cytochrome c in the absence of dATP but does not form the oligomeric complex. However, when dATP is added to the cytochrome c-bound Apaf-1 complex, complete oligomerization occurs, suggesting that oligomerization is driven by hydrolysis of dATP. This was supported by the observation that ATP, but not the nonhydrolyzable adenosine 5'-O (thiotriphosphate), can induce oligomerization of the Apaf-1-cytochrome c complex. Like the spontaneously oligomerizing Apaf-530, which lacks its WD-40 domain, the oligomeric full-length Apaf-1-cytochrome c complex can bind and process procaspase-9 in the absence of additional dATP or cytochrome c. However, unlike the truncated Apaf-530 complex, the full-length Apaf-1 complex can release the mature caspase-9 after processing. Once released, mature caspase-9 can process procaspase-3, setting into motion the caspase cascade. These observations indicate that cytochrome c and dATP are required for oligomerization of Apaf-1 and suggest that the WD-40 domain plays an important role in oligomerization of full-length Apaf-1 and the release of mature caspase-9 from the Apaf-1 oligomeric complex. PMID- 10364242 TI - CLAP, a novel caspase recruitment domain-containing protein in the tumor necrosis factor receptor pathway, regulates NF-kappaB activation and apoptosis. AB - Molecules that regulate NF-kappaB activation play critical roles in apoptosis and inflammation. We describe the cloning of the cellular homolog of the equine herpesvirus-2 protein E10 and show that both proteins regulate apoptosis and NF kappaB activation. These proteins were found to contain N-terminal caspase recruitment domains (CARDs) and novel C-terminal domains (CTDs) and were therefore named CLAPs (CARD-like apoptotic proteins). The cellular and viral CLAPs induce apoptosis downstream of caspase-8 by activating the Apaf-1-caspase-9 pathway and activate NF-kappaB by acting upstream of the NF-kappaB-inducing kinase, NIK, and the IkB kinase, IKKalpha. Deletion of either the CARD or the CTD domain inhibits both activities. The CARD domain was found to be important for homo- and heterodimerization of CLAPs. Substitution of the CARD domain with an inducible FKBP12 oligomerization domain produced a molecule that can induce NF kappaB activation, suggesting that the CARD domain functions as an oligomerization domain, whereas the CTD domain functions as the effector domain in the NF-kappaB activation pathway. Expression of the CARD domain of human CLAP abrogates tumor necrosis factor-alpha-induced NF-kappaB activation, suggesting that cellular CLAP plays an essential role in this pathway of NF-kappaB activation. PMID- 10364243 TI - Genetic analyses of proteolysis, hemoglobin binding, and hemagglutination of Porphyromonas gingivalis. Construction of mutants with a combination of rgpA, rgpB, kgp, and hagA. AB - Porphyromonas gingivalis produces arginine-specific cysteine proteinase (Arg gingipain, RGP) and lysine-specific cysteine proteinase (Lys-gingipain, KGP) in the extracellular and cell-associated forms. Two separate genes (rgpA and rgpB) and a single gene (kgp) have been found to encode RGP and KGP, respectively. We constructed rgpA rgpB kgp triple mutants by homologous recombination with cloned rgp and kgp DNA interrupted by drug resistance gene markers. The triple mutants showed no RGP or KGP activity in either cell extracts or culture supernatants. The culture supernatants of the triple mutants grown in a rich medium had no proteolytic activity toward bovine serum albumin or gelatin derived from human type I collagen. Moreover, the mutants did not grow in a defined medium containing bovine serum albumin as the sole carbon/energy source. These results indicate that the proteolytic activity of P. gingivalis toward bovine serum albumin and gelatin derived from human type I collagen appears to be attributable to RGP and KGP. The hemagglutinin gene hagA of P. gingivalis possesses the adhesin domain regions responsible for hemagglutination and hemoglobin binding that are also located in the C-terminal regions of rgpA and kgp. A rgpA kgp hagA triple mutant constructed in this study exhibited no hemagglutination using sheep erythrocytes or hemoglobin binding activity, as determined by a solid-phase binding assay with horseradish peroxidase-conjugated human hemoglobin, indicating that the adhesin domains seem to be particularly important for P. gingivalis cells to agglutinate erythrocytes and bind hemoglobin, leading to heme acquisition. PMID- 10364244 TI - Temporal association of the N- and O-linked glycosylation events and their implication in the polarized sorting of intestinal brush border sucrase isomaltase, aminopeptidase N, and dipeptidyl peptidase IV. AB - The temporal association between O-glycosylation and processing of N-linked glycans in the Golgi apparatus as well as the implication of these events in the polarized sorting of three brush border proteins has been the subject of the current investigation. O-Glycosylation of pro-sucrase-isomaltase (pro-SI), aminopeptidase N (ApN), and dipeptidyl peptidase IV (DPPIV) is drastically reduced when processing of the mannose-rich N-linked glycans is blocked by deoxymannojirimycin, an inhibitor of the Golgi-located mannosidase I. By contrast, O-glycosylation is not affected in the presence of swainsonine, an inhibitor of Golgi mannosidase II. The results indicate that removal of the outermost mannose residues by mannosidase I from the mannose-rich N-linked glycans is required before O-glycosylation can ensue. On the other hand, subsequent mannose residues in the core chain impose no sterical constraints on the progression of O-glycosylation. Reduction or modification of N- and O glycosylation do not affect the transport of pro-SI, ApN, or DPPIV to the cell surface per se. However, the polarized sorting of two of these proteins, pro-SI and DPPIV, to the apical membrane is substantially altered when O-glycans are not completely processed, while the sorting of ApN is not affected. The processing of N-linked glycans, on the other hand, has no influence on sorting of all three proteins. The results indicate that O-linked carbohydrates are at least a part of the sorting mechanism of pro-SI and DPPIV. The sorting of ApN implicates neither O-linked nor N-linked glycans and is driven most likely by carbohydrate independent mechanisms. PMID- 10364245 TI - C-reactive protein inhibits chemotactic peptide-induced p38 mitogen-activated protein kinase activity and human neutrophil movement. AB - Serum levels of the acute-phase reactant, C-reactive protein (CRP), increase dramatically during acute inflammatory episodes. CRP inhibits migration of neutrophils toward the chemoattractant, f-Met-Leu-Phe (fMLP) and therefore acts as an anti-inflammatory agent. Since tyrosine kinases are involved in neutrophil migration and CRP has been shown to decrease phosphorylation of some neutrophil proteins, we hypothesized that CRP inhibits neutrophil chemotaxis via inhibition of MAP kinase activity. The importance of p38 MAP kinase in neutrophil movement was determined by use of the specific p38 MAP kinase inhibitor, SB203580. CRP and SB203580 both blocked random and fMLP-directed neutrophil movement in a concentration-dependent manner. Additionally, extracellular signal-regulated MAP kinase (ERK) was not involved in fMLP-induced neutrophil movement as determined by use of the MEK-specific inhibitor, PD98059. Blockade of ERK with PD98059 did not inhibit chemotaxis nor did it alter the ability of CRP or SB203580 to inhibit fMLP-induced chemotaxis. More importantly, CRP inhibited fMLP-induced p38 MAP kinase activity in a concentration-dependent manner as measured by an in vitro kinase assay. Impressively, CRP-mediated inhibition of p38 MAP kinase activity correlated with CRP-mediated inhibition of fMLP-induced chemotaxis (r = -0.7144). These data show that signal transduction through p38 MAP kinase is necessary for neutrophil chemotaxis and that CRP intercedes through this pathway in inhibiting neutrophil movement. PMID- 10364246 TI - Distinct functions of eukaryotic translation initiation factors eIF1A and eIF3 in the formation of the 40 S ribosomal preinitiation complex. AB - We have used an in vitro translation initiation assay to investigate the requirements for the efficient transfer of Met-tRNAf (as Met-tRNAf.eIF2.GTP ternary complex) to 40 S ribosomal subunits in the absence of mRNA (or an AUG codon) to form the 40 S preinitiation complex. We observed that the 17-kDa initiation factor eIF1A is necessary and sufficient to mediate nearly quantitative transfer of Met-tRNAf to isolated 40 S ribosomal subunits. However, the addition of 60 S ribosomal subunits to the 40 S preinitiation complex formed under these conditions disrupted the 40 S complex resulting in dissociation of Met-tRNAf from the 40 S subunit. When the eIF1A-dependent preinitiation reaction was carried out with 40 S ribosomal subunits that had been preincubated with eIF3, the 40 S preinitiation complex formed included bound eIF3 (40 S.eIF3. Met tRNAf.eIF2.GTP). In contrast to the complex lacking eIF3, this complex was not disrupted by the addition of 60 S ribosomal subunits. These results suggest that in vivo, both eIF1A and eIF3 are required to form a stable 40 S preinitiation complex, eIF1A catalyzing the transfer of Met-tRNAf.eIF2.GTP to 40 S subunits, and eIF3 stabilizing the resulting complex and preventing its disruption by 60 S ribosomal subunits. PMID- 10364247 TI - Protease-resistant and detergent-insoluble prion protein is not necessarily associated with prion infectivity. AB - PrPSc, an abnormal isoform of PrPC, is the only known component of the prion, an agent causing fatal neurodegenerative disorders such as bovine spongiform encephalopathy (BSE) and Creutzfeldt-Jakob disease (CJD). It has been postulated that prion diseases propagate by the conversion of detergent-soluble and protease sensitive PrPC molecules into protease-resistant and insoluble PrPSc molecules by a mechanism in which PrPSc serves as a template. We show here that the chemical chaperone dimethyl sulfoxide (Me2SO) can partially inhibit the aggregation of either PrPSc or that of its protease-resistant core PrP27-30. Following Me2SO removal by methanol precipitation, solubilized PrP27-30 molecules aggregated into small and amorphous structures that did not resemble the rod configuration observed when scrapie brain membranes were extracted with Sarkosyl and digested with proteinase K. Interestingly, aggregates derived from Me2SO-solubilized PrP27 30 presented less than 1% of the prion infectivity obtained when the same amount of PrP27-30 in rods was inoculated into hamsters. These results suggest that the conversion of PrPC into protease-resistant and detergent-insoluble PrP molecules is not the only crucial step in prion replication. Whether an additional requirement is the aggregation of newly formed proteinase K-resistant PrP molecules into uniquely structured aggregates remains to be established. PMID- 10364248 TI - CCAAT enhancer-binding protein beta and GATA-4 binding regions within the promoter of the steroidogenic acute regulatory protein (StAR) gene are required for transcription in rat ovarian cells. AB - Steroidogenic acute regulatory protein (StAR) is a vital accessory protein required for biosynthesis of steroid hormones from cholesterol. The present study shows that in primary granulosa cells from prepubertal rat ovary, StAR transcript and protein are acutely induced by gonadotropin (FSH). To determine the sequence elements required for hormone inducibility of the StAR promoter, truncated regions of the -1002/+6 sequence of the mouse gene were ligated to pCAT-Basic plasmid and transfected by electroporation to freshly prepared cells. FSH inducibility determined over a 6-h incubation was 10-40-fold above basal levels of chloramphenicol acetyltransferase activity. These functional studies, supported by electrophoretic mobility shift assays indicated that two sites were sufficient for transcription of the StAR promoter constructs: a non-consensus binding sequence (-81/-72) for CCAAT enhancer-binding protein beta (C/EBPbeta) and a consensus motif for GATA-4 binding (-61/-66). Western analyses showed that GATA-4 is constitutively expressed in the granulosa cells, while all isoforms of C/EBPbeta were markedly inducible by FSH. Site-directed mutations of both binding sequences practically ablated both basal and hormone-driven chloramphenicol acetyltransferase activities to less than 5% of the parental -96/+6 construct. Unlike earlier notions, elimination of potential binding sites for steroidogenic factor-1, a well known tissue-specific transcription factor, did not impair StAR transcription. Consequently, we propose that C/EBPbeta and GATA-4 represent a novel combination of transcription factors capable of conferring an acute response to hormones upon their concomitant binding to the StAR promoter. PMID- 10364249 TI - Insulin-induced early growth response gene (Egr-1) mediates a short term repression of rat malic enzyme gene transcription. AB - In this report we have studied insulin regulation of malic enzyme (ME) gene transcription in rat H-35 hepatoma cells and localized the insulin-responsive region of the ME promoter between positions -177 and -102. This region contains a putative insulin response element (IRE-II). When nuclear extracts from untreated or insulin-treated H-35 cells were incubated with IRE-II, transcription factors Sp1 and Sp3 were observed to bind constitutively to this element, whereas insulin induces the quick and transient binding of an insulin response factor. This induction requires de novo protein synthesis. Competition and supershift assays demonstrated that the insulin response factor is the immediate-early gene Egr-1. In vitro assays revealed that Egr-1 displaces Sp1 from its binding site in IRE II. Insulin induces Egr-1 mRNA, with a time course pattern that corresponds perfectly to the Egr-1 binding to IRE-II. This induction depends on the activation of mitogen-activated protein (MAP) kinase, and it is phosphatidylinositol 3-kinase-independent, as demonstrated with specific inhibitors for both pathways. By cotransfecting the wild-type or a dominant negative Ras, an upstream regulator of MAP kinase, we show that Ras inhibits ME promoter activity. Furthermore, overexpression of Egr-1 in H-35 cells represses the ME gene promoter in a dose-dependent manner. These results suggest that insulin induces a quick, transient, and Ras/MAP kinase-dependent activation of Egr-1 which leads to a transient repression of ME gene transcription. On a late phase, insulin would activate a different, Egr-1-independent pathway, which would result in activation of the ME gene. PMID- 10364250 TI - Signal relay by retinoic acid receptors alpha and beta in the retinoic acid induced expression of insulin-like growth factor-binding protein-3 in breast cancer cells. AB - Neither retinoic acid receptor-beta (RARbeta) nor insulin-like growth factor binding protein-3 (IGFBP-3) is expressed in breast cancer cell line MCF-7. The expression of both proteins can be induced in response to all-trans-retinoic acid (atRA). By using an RARalpha-selective antagonist (Ro 41-5253), we demonstrated that RARbeta expression was induced by atRA through an RARalpha-dependent signaling pathway and that RARbeta induction was correlated with IGFBP-3 induction. However, MCF-7 cells transfected with sense RARbeta cDNA expressed IGFBP-3 even in the presence of the RARalpha-selective antagonist Ro 41-5253. On the other hand, antisense RARbeta cDNA transfection of MCF-7 cells blocked atRA induced IGFBP-3 expression, indicating that RARbeta is directly involved in the mediation of IGFBP-3 induction by atRA. Induction of IGFBP-3 expression by atRA occurs at the transcriptional level, as measured by nuclear run-on assays. Finally, we showed that atRA-induced IGFBP-3 is functionally active in modulating the growth-promoting effect of IGF-I. These experiments indicate that RARalpha and RARbeta, both individually and together, are important in mammary gland homeostasis and breast cancer development. By linking IGFBP-3 to RARbeta, our experiments define the signal intersection between the retinoid and IGF systems in cell growth regulation and explain why loss of RARbeta might be critical in breast cancer carcinogenesis/progression. PMID- 10364251 TI - Evidence for covalent attachment of diphytanylglyceryl phosphate to the cell surface glycoprotein of Halobacterium halobium. AB - In a previous study, we demonstrated the occurrence of novel proteins modified with a diphytanylglyceryl group in thioether linkage in Halobacterium halobium (Sagami, H., Kikuchi, A., and Ogura, K. (1995) J. Biol. Chem. 270, 14851-14854). In this study, we further investigated protein isoprenoid modification in this halobacterium using several radioactive tracers such as [3H]geranylgeranyl diphosphate. One of the radioactive bands observed on SDS-polyacrylamide gel electrophoresis corresponded to a periodic acid-Schiff stain-positive protein (200 kDa). Radioactive and periodic acid-Schiff stain-positive peptides (28 kDa) were obtained by trypsin digestion of the labeled proteins. The radioactive materials released by acid treatment of the peptides showed a similar mobility to dolichyl (C55) phosphate on a normal-phase thin-layer plate. However, radioactive hydrolysates obtained by acid phosphatase treatment co-migrated not with dolichol (C55-65), but with diphytanylglycerol on both reverse- and normal-phase thin layer plates. The mass spectrum of the hydrolysate was also coincident with that of diphytanylglycerol. The partial amino acid sequences of the 28-kDa peptides were found in a fragment (amino acids 731-816) obtainable by trypsin cleavage of the known cell-surface glycoprotein of this halobacterium. These results indicate that the cell-surface glycoprotein (200 kDa) is modified with diphytanylglyceryl phosphate. PMID- 10364252 TI - Functional characterization of multiple transactivating elements in beta-catenin, some of which interact with the TATA-binding protein in vitro. AB - beta-Catenin, a member of the family of Armadillo repeat proteins, plays a dual role in cadherin-mediated cell adhesion and in signaling by Wnt growth factors. Upon Wnt stimulation beta-catenin undergoes nuclear translocation and serves as transcriptional coactivator of T cell factor DNA-binding proteins. Previously the transactivation potential of different portions of beta-catenin has been demonstrated, but the precise location of transactivating elements has not been established. Also, the mechanism of transactivation by beta-catenin and the molecular basis for functional differences between beta-catenin and the closely related proteins Armadillo and Plakoglobin are poorly understood. Here we have used a yeast system for the detailed characterization of the transactivation properties of beta-catenin. We show that its transactivation domains possess a modular structure, consist of multiple subelements that cover broad regions at its N and C termini, and extend considerably into the Armadillo repeat region. Compared with beta-catenin the N termini of Plakoglobin and Armadillo have different transactivation capacities that may explain their distinct signaling properties. Furthermore, transactivating elements of beta-catenin interact specifically and directly with the TATA-binding protein in vitro providing further evidence that a major function of beta-catenin during Wnt signaling is to recruit the basal transcription machinery to promoter regions of Wnt target genes. PMID- 10364253 TI - Biochemical analysis of the receptor for ubiquitin-like polypeptide. AB - Monoclonal nonspecific suppressor factor (MNSF), a lymphokine produced by murine T cell hybridoma, possesses pleiotrophic antigen-nonspecific suppressive functions. A cDNA clone encoding MNSF-beta, an isoform of the MNSF, has been isolated and characterized. MNSF-beta cDNA encodes a fusion protein consisting of a ubiquitin-like segment (Ubi-L) and ribosomal protein S30. Ubi-L appears to be cleaved from the ribosomal protein and released extracellularly in association with T cell receptor-like polypeptide. In the current study we have characterized the biochemical nature of the Ubi-L receptor on D.10 G4.1, a murine T helper clone type 2. Biotinylated Ubi-L bound preferentially to concanavalin A stimulated but not to unstimulated D.10 cells. Detergent-extracted membrane proteins were applied to an immobilized Ubi-L column. SDS-polyacrylamide gel electrophoresis of eluted fraction revealed a band of Mr = 82,000. Biotinylated Ubi-L specifically recognized this band, confirming that the 82-kDa protein is the Ubi-L receptor. A complex of Mr = 90,000 was visualized by immunoprecipitation of 125I-Ubi-L cross-linked to the purified receptor followed by SDS-polyacrylamide gel electrophoresis and autoradiography. In addition, a 105 kDa protein was coimmunoprecipitated by anti-Ubi-L receptor (82-kDa polypeptide) antibody, indicative of the association of this protein with the Ubi-L receptor complex. Amino acid sequence analysis of the 82-kDa polypeptide revealed that the Ubi-L receptor may be a member of a cytokine receptor family. PMID- 10364254 TI - The African swine fever virus prenyltransferase is an integral membrane trans geranylgeranyl-diphosphate synthase. AB - In a previous study, it was shown that the protein encoded by the gene B318L of African swine fever virus (ASFV) is a trans-prenyltransferase that catalyzes in vitro the condensation of farnesyl diphosphate and isopentenyl diphosphate to synthesize geranylgeranyl diphosphate and longer chain prenyl diphosphates (Alejo, A., Yanez, R. J., Rodriguez, J. M., Vinuela, E., and Salas, M. L. (1997) J. Biol. Chem. 272, 9417-9423). To investigate the in vivo function of the viral enzyme, we have determined, in this work, its subcellular localization and activity in cell extracts. Two systems were used in these studies: cells infected with ASFV and cells infected with a recombinant pseudo-Sindbis virus carrying the complete B318L gene. In this latter system, the trans-prenyltransferase was found to colocalize with the endoplasmic reticulum marker protein-disulfide isomerase, whereas in cells infected with ASFV, the viral enzyme was present in cytoplasmic viral assembly sites, associated with precursor viral membranes derived from the endoplasmic reticulum. In addition, after subcellular fractionation, the viral enzyme partitioned into the membrane fraction. Extraction of membrane proteins with alkaline carbonate and Triton X-114 indicated that the ASFV enzyme behaved as an integral membrane protein. The membrane enzyme synthesized predominantly all-trans-geranylgeranyl diphosphate from farnesyl diphosphate and isopentenyl diphosphate. These results indicate that the viral B318L protein is a trans geranylgeranyl-diphosphate synthase, being the only enzyme of this type that is known to have a membrane localization. PMID- 10364255 TI - Identification of a general transcription factor TFIIAalpha/beta homolog selectively expressed in testis. AB - In this paper we describe the isolation of a cDNA that encodes a human TFIIAalpha/beta-like factor (ALF). The open reading frame of ALF predicts a protein of 478 amino acids that contains characteristic N- and C-terminal conserved domains separated by an internal nonconserved domain. In addition, a rare ALF-containing cDNA, which possesses an extended N terminus (Stoned B/TFIIAalpha/beta-like factor; SALF) has also been identified. The results of Northern and dot blot analyses show that ALF is expressed almost exclusively in testis; in contrast, TFIIAalpha/beta and TFIIAgamma are enriched in testis but are also widely expressed in other human tissues. Recombinant ALF (69 kDa) and TFIIAgamma (12 kDa) polypeptides produced in Escherichia coli form an ALF/gamma complex that can stabilize TBP-TATA interactions in an electrophoretic mobility shift assay. The ALF/gamma complex is also able to restore transcription from the adenovirus major late promoter using HeLa cell nuclear extracts that have been depleted of TFIIA. Overall, the data show that ALF is a functional homolog of human general transcription factor TFIIAalpha/beta that may be uniquely important to testis biology. PMID- 10364256 TI - Functional early endosomes are required for maturation of major histocompatibility complex class II molecules in human B lymphoblastoid cells. AB - Major histocompatibility complex (MHC) class II molecules are targeted together with their invariant chain (Ii) chaperone from the secretory pathway to the endocytic pathway. Within the endosome/lysosome system, Ii must be degraded to enable peptide capture by MHC class II molecules. It remains controversial exactly which route or routes MHC class II/Ii complexes take to reach the sites of Ii processing and peptide loading. We have asked whether early endosomes are required for successful maturation of MHC class II molecules by using an in situ peroxidase/diaminobenzidine compartment ablation technique. Cells whose early endosomes were selectively ablated using transferrin-horseradish peroxidase conjugates fail to mature their newly synthesized MHC class II molecules. We show that whereas transport of secretory Ig through the secretory pathway is virtually normal in the ablated cells, newly synthesized MHC class II/Ii complexes never reach compartments capable of processing Ii. These results strongly suggest that the transport of the bulk of newly synthesized MHC class II molecules through early endosomes is obligatory and that direct input into later endosomes/lysosomes does not take place. PMID- 10364257 TI - Phosphoinositide 3-kinase-dependent and -independent activation of the small GTPase Rac2 in human neutrophils. AB - The small GTPase Rac participates in various cellular events such as cytoskeletal reorganization. It has remained, however, largely unknown about intracellular signaling pathways for Rac activation because of the lack of a simple and reliable assay to estimate the activation. Here we describe a novel method to detect the GTP-bound, active Rac in cells by pulling it down with the Rac-binding domain of the protein kinase PAK. Experiments using this method reveal that stimulation of human neutrophils with the Gi-coupled receptor agonists N-formyl methionyl-leucyl-phenylalanine (fMLP) and leukotriene B4 (LTB4) leads to a rapid and transient increase in the GTP-bound state of Rac2, whereas phorbol myristate acetate (PMA) causes a slow but more sustained activation of Rac2. Pretreatment of cells with pertussis toxin results in defective activation of Rac2 in response to fMLP and LTB4, indicating that coupling of the receptors to Gi plays a crucial role in the activation. Furthermore, the phosphoinositide 3-kinase (PI3K) inhibitors wortmannin and LY294002 block Rac2 activation elicited by the receptor agonists, but not that by PMA. Thus the Gi-coupled receptors likely mediate Rac2 activation via PI3K, whereas PMA activates Rac2 in a PI3K-independent manner. PMID- 10364258 TI - Lipopolysaccharide inhibits virus-mediated induction of interferon genes by disruption of nuclear transport of interferon regulatory factors 3 and 7. AB - We have studied the effects of lipopolysaccharide (LPS) on the Newcastle disease virus (NDV)-mediated induction of cytokine genes expression. Raw cells treated with LPS before or after virus infection showed down-regulation in the expression of interferon A and, to a lesser extent, interferon B genes. In contrast, induction of the interleukin (IL)-6 gene was enhanced. The effects of LPS were not a result of the suppression of virus replication, because the transcription of viral nucleocapsid gene was not affected. Consistent with these findings, LPS also suppressed the NDV-mediated induction of chloramphenicol acetyltransferase reporter gene driven by murine interferon A4 promoter in a transient transfection assay. Furthermore, LPS inhibited virus-mediated phosphorylation of interferon regulatory factor (IRF)-3 and the consequent translocation of IRF-3 from cytoplasm to nucleus. The LPS-mediated inhibition of IFNA gene expression was much weaker in infected Raw cells that constitutively overexpressed IRF-3. The nuclear translocation of IRF-7 in infected cells was also inhibited by LPS. These data suggest that LPS down-regulates the virus-mediated induction of IFNA genes by post-translationally targeting the IRF-3 and IRF-7 proteins. PMID- 10364260 TI - Human alveolar macrophages are markedly deficient in REF-1 and AP-1 DNA binding activity. AB - Although many functions of human alveolar macrophages are altered compared with their precursor cell, the blood monocyte (monocyte), the reason(s) for these functional changes have not been determined. We recently reported that human alveolar macrophages do not express AP-1 DNA binding activity (Monick, M. M., Carter, A. B., Gudmundsson, G., Geist, L. J., and Hunninghake, G. W. (1998) Am. J. Physiol. 275, L389-L397). To determine why alveolar macrophages do not express AP-1 DNA binding activity, we first showed that there was not a decrease in expression of the FOS and JUN proteins that make up the AP-1 complex. There was, however, a significant difference in the amounts of the nuclear protein, REF-1 (which regulates AP-1 DNA binding by altering the redox status of FOS and JUN proteins), in alveolar macrophages compared with monocytes. In addition, in vitro differentiation of monocytes to a macrophage-like cell resulted in decreased amounts of REF-1. Finally, addition of REF-1 from activated monocytes to alveolar macrophage nuclear proteins resulted in a marked increase in AP-1 DNA binding. These studies strongly suggest that the process of differentiation of monocytes into alveolar macrophages is associated with a loss of REF-1 and AP-1 activity. This observation may explain, in part, some of the functional differences observed for alveolar macrophages compared with monocytes. PMID- 10364259 TI - Insulin receptor substrate 1-induced inhibition of endoplasmic reticulum Ca2+ uptake in beta-cells. Autocrine regulation of intracellular ca2+ homeostasis and insulin secretion. AB - To understand the role of the insulin receptor pathway in beta-cell function, we have generated stable beta-cells (betaIRS1-A) that overexpress by 2-fold the insulin receptor substrate-1 (IRS-1) and compared them to vector-expressing controls. IRS-1 overexpression dramatically increased basal cytosolic Ca2+ levels from 81 to 278 nM, but it did not affect Ca2+ response to glucose. Overexpression of the insulin receptor also caused an increase in cytosolic Ca2+. Increased cytosolic Ca2+ was due to inhibition of Ca2+ uptake by the endoplasmic reticulum, because endoplasmic reticulum Ca2+ uptake and content were reduced in betaIRS1-A cells. Fractional insulin secretion was significantly increased 2-fold, and there was a decrease in betaIRS1-A insulin content and insulin biosynthesis. Steady state insulin mRNA levels and glucose-stimulated ATP were unchanged. High IRS-1 levels also reduced beta-cell proliferation. These data demonstrate a direct link between the insulin receptor signaling pathway and the Ca2+-dependent pathways regulating insulin secretion of beta-cells. We postulate that during regulated insulin secretion, released insulin binds the beta-cell insulin receptor and activates IRS-1, thus further increasing cytosolic Ca2+ by reducing Ca2+ uptake. We suggest the existence of a novel pathway of autocrine regulation of intracellular Ca2+ homeostasis and insulin secretion in the beta-cell of the endocrine pancreas. PMID- 10364261 TI - Effects of mutations of a phosphorylation site in an exposed loop in acidic fibroblast growth factor. AB - Acidic fibroblast growth factor (aFGF) contains a phosphorylation site recognized by protein kinase C. A non-mitogenic mutant growth factor is devoid of this phosphorylation site. We have changed amino acids in and close to the phosphorylation site and studied the consequences of this for binding of the growth factor to high affinity receptors as well as to heparin. We have also studied the ability of the mutants to stimulate DNA synthesis and cell proliferation as well as phosphorylation of mitogen-activated protein kinase and the ability of the growth factor mutants to be transported to the nucleus. The results indicate that while the mutations strongly affect the ability of the growth factor to bind to heparin, they do not affect much the binding to the specific FGF receptors, activation of mitogen-activated protein kinase or transport of the growth factor to the nucleus. The mutations affect to various extents the ability of the growth factor to stimulate DNA synthesis and to induce cell multiplication. We find that phosphorylation of aFGF is not required for mitogenic activity. The data suggest that altered interaction of the growth factor with a cellular component different from the receptor, possibly a component in the nucleus, is the reason for the different mitogenicity of the different growth factor mutants. PMID- 10364262 TI - A vanadium and iron cluster accumulates on VnfX during iron-vanadium-cofactor synthesis for the vanadium nitrogenase in Azotobacter vinelandii. AB - The vnf-encoded nitrogenase from Azotobacter vinelandii contains an iron-vanadium cofactor (FeV-co) in its active site. Little is known about the synthesis pathway of FeV-co, other than that some of the gene products required are also involved in the synthesis of the iron-molybdenum cofactor (FeMo-co) of the widely studied molybdenum-dinitrogenase. We have found that VnfX, the gene product of one of the genes contained in the vnf-regulon, accumulates iron and vanadium in a novel V-Fe cluster during synthesis of FeV-co. The electron paramagnetic resonance (EPR) and metal analyses of the V-Fe cluster accumulated on VnfX are consistent with a VFe7 8Sx precursor of FeV-co. The EPR spectrum of VnfX with the V-Fe cluster bound strongly resembles that of isolated FeV-co and a model VFe3S4 compound. The V-Fe cluster accumulating on VnfX does not contain homocitrate. No accumulation of V Fe cluster on VnfX was observed in strains with deletions in genes known to be involved in the early steps of FeV-co synthesis, suggesting that it corresponds to a precursor of FeV-co. VnfX purified from a nifB strain incapable of FeV-co synthesis has a different electrophoretic mobility in native anoxic gels than does VnfX, which has the V-Fe cluster bound. NifB-co, the Fe and S precursor of FeMo-co (and presumably FeV-co), binds to VnfX purified from the nifB strain, producing a shift in its electrophoretic mobility on anoxic native gels. The data suggest that a precursor of FeV-co that contains vanadium and iron accumulates on VnfX, and thus, VnfX is involved in the synthesis of FeV-co. PMID- 10364263 TI - Insulin receptor substrate 3 is not essential for growth or glucose homeostasis. AB - The insulin receptor substrates (IRS) 1 and 2 are required for normal growth and glucose homeostasis in mice. To determine whether IRS-3, a recently cloned member of the IRS family, is also involved in the regulation of these, we have generated mice with a targeted disruption of the IRS-3 gene and characterized them. Compared with wild-type mice, the IRS-3-null mice showed normal body weight throughout development, normal blood glucose levels in the fed and fasted state and following an oral glucose bolus, and normal fed and fasted plasma insulin levels. IRS-3 is most abundant in adipocytes and is tyrosine-phosphorylated in response to insulin in these cells. Therefore, isolated adipocytes were analyzed for changes in insulin effects. Insulin-stimulated glucose transport in the adipocytes from the IRS-3-null mice was the same as in wild-type cells. The extent of tyrosine phosphorylation of IRS-1/2 following insulin stimulation was similar in adipocytes from IRS-3-null and wild-type mice, and the insulin-induced association of tyrosine-phosphorylated IRS-1/2 with phosphatidylinositol 3-kinase and SHP-2 was not detectably increased by IRS-3 deficiency. Thus, IRS-3 was not essential for normal growth, glucose homeostasis, and glucose transport in adipocytes, and in its absence no significant compensatory augmentation of insulin signaling through IRS-1/2 was evident. PMID- 10364264 TI - Sustained mitogen-activated protein kinase (MAPK) and cytoplasmic phospholipase A2 activation by macrophage migration inhibitory factor (MIF). Regulatory role in cell proliferation and glucocorticoid action. AB - Macrophage migration inhibitory factor (MIF) is an important pro-inflammatory mediator with the unique ability to counter-regulate the inhibitory effects of glucocorticoids on immune cell activation. MIF is released from cells in response to glucocorticoids, certain pro-inflammatory stimuli, and mitogens and acts to regulate glucocorticoid action on the ensuing inflammatory response. To gain insight into the molecular mechanism of MIF action, we have examined the role of MIF in the proliferation and intracellular signaling events of the well characterized, NIH/3T3 fibroblast cell line. Both endogenously secreted and exogenously added MIFs stimulate the proliferation of NIH/3T3 cells, and this response is associated with the activation of the p44/p42 extracellular signal regulated (ERK) mitogen-activated protein kinases (MAP). The MIF-induced activation of these kinases was sustained for a period of at least 24 h and was dependent upon protein kinase A activity. We further show that MIF regulates cytosolic phospholipase A2 activity via a protein kinase A and ERK dependent pathway and that the glucocorticoid suppression of cytokine-induced cytoplasmic phospholipase A2 activity and arachidonic acid release can be reversed by the addition of recombinant MIF. These studies indicate that the sustained activation of p44/p42 MAP kinase and subsequent arachidonate release by cytoplasmic phospholipase A2 are important features of the immunoregulatory and intracellular signaling events initiated by MIF and provide the first insight into the mechanisms that underlie the pro-proliferative and inflammatory properties of this mediator. PMID- 10364265 TI - Continuous autotropic signaling by membrane-expressed tumor necrosis factor. AB - Tumor necrosis factor (TNF) exists in two bioactive forms, the membrane integrated form and the proteolytically derived soluble cytokine. Cells that produce TNF are often responsive to TNF, allowing autocrine/juxtacrine feedback loops. However, whether the membrane form of TNF is involved in such regulatory circuits is unclear. Here we demonstrate that HeLa cells, expressing a permanently membrane-integrated mutant form of TNF, constitutively express TNF.TNF receptor complexes at their cell surface. These cells show a permanent activation of the transcription factor NF-kappaB, exert constitutive p38 mitogen activated protein kinase activity, and produce high amounts of interleukin-6. In parallel, transmembrane TNF-expressing HeLa cells display high sensitivity to cycloheximide or interferon-gamma, similar to untransfected cells treated with these agents in combination with sTNF. Moreover, cycloheximide-induced apoptosis in transmembrane TNF transfectants can be blocked by the caspase inhibitor zVAD fmk and does not necessarily need cell to cell contact, indicating a critical role of constitutive autotropic signaling of TNF.TNF receptor complexes. These data demonstrate that autotropic signaling loops of membrane TNF can exist, which may be of importance for cells that express both TNF and TNF receptors, such as T lymphocytes, macrophages, and endothelial cells. PMID- 10364266 TI - Comparison of the effects on secretion in chromaffin and PC12 cells of Rab3 family members and mutants. Evidence that inhibitory effects are independent of direct interaction with Rabphilin3. AB - The Rab class of low molecular weight GTPases has been implicated in the regulation of vesicular trafficking between membrane compartments in eukaryotic cells. The Rab3 family consisting of four highly homologous isoforms is associated with secretory granules and synaptic vesicles. Many different types of experiments indicate that Rab3a is a negative regulator of exocytosis and that its GTP-bound form interacts with Rabphilin3, a possible effector. Overexpression of Rabphilin3 in chromaffin cells enhances secretion. We have investigated the expression, localization, and effects on secretion of the various members of the Rab3 family in bovine chromaffin and PC12 cells. We found that Rab3a, Rab3b, Rab3c, and Rab3d are expressed to varying degrees in PC12 cells and in a fraction enriched in chromaffin granule membranes from the adrenal medulla. Immunocytochemistry revealed that all members of the family when overexpressed in PC12 cells localize to secretory granules. Binding constants for the interaction of the GTP-bound forms of Rab3a, Rab3b, Rab3c, and Rab3d with Rabphilin3 were comparable (Kd = 10-20 nM). Overexpression of each of the four members of the Rab3 family inhibited secretion. Mutations in Rab3a were identified that strongly impaired the ability of the GTP-bound form to interact with Rabphilin3. The mutated proteins inhibited secretion similarly to wild type Rab3a. Although Rab3a and Rabphilin3 are located on the same secretory granule or secretory vesicle and interact both in vitro and in situ, it is concluded that the inhibition of secretion by overexpression of Rab3a is unrelated to its ability to interact with Rabphilin3. PMID- 10364267 TI - Receptor interacting protein RIP140 inhibits both positive and negative gene regulation by glucocorticoids. AB - Recent development in the field of gene regulation by nuclear receptors (NRs) have identified a role for cofactors in transcriptional control. While some of the NR-associated proteins serve as coactivators, the effect of the receptor interacting protein 140 (RIP140) on NR transcriptional responses is complex. In this report we have studied the effect of RIP140 on gene regulation by the glucocorticoid receptor (GR). We demonstrate that RIP140 antagonized all GR mediated responses tested, which included activation through classical GRE, the synergistic effects of glucocorticoids on AP-1 and Pbx1/HOXB1 responsive elements, as well as gene repression through a negative GRE and cross-talk with NF-kappaB (RelA). This involved the ligand-binding domain of the GR and did not occur when the GR was bound to the antagonist RU486. The strong repressive effect of RIP140 was restricted to glucocorticoid-mediated responses in as much as it slightly increased signaling through the RelA and the Pit-1/Pbx proteins and only slightly repressed signaling through the Pbx1/HOXB1 and AP-1 proteins, excluding general squelching as a mechanism. Instead, this suggests that RIP140 acts as a direct inhibitor of GR function. In line with a direct effect of RIP140 on the GR, we demonstrate a GR-RIP140 interaction in vitro by a glutathione S transferase-pull down assay. Furthermore, the repressive effect of RIP140 could partially be overcome by overexpression of the coactivator TIF2, which involved a competition between TIF2 and RIP140 for binding to the GR. PMID- 10364268 TI - Retinoic acid confers resistance to p53-dependent apoptosis in SH-SY5Y neuroblastoma cells by modulating nuclear import of p53. AB - Many cell lines derived from neuroblastoma (NB) carry the wild-type p53 gene with a p53-dependent apoptotic pathway that is responsive to DNA damaging agents. A recent study has demonstrated that retinoic acid (RA) pretreatment of NB cells promotes chemoresistance to apoptosis induced by chemotherapeutic agents. We examine here the possible contribution of the p53 pathway to the chemoresistance response associated with the RA treatment in NB cells. Upon treatment with RA (1 10 microM) for 4 days, the human NB cells, SH-SY5Y, developed resistance selectively to p53-dependent apoptotic stimuli including gamma-irradiation, etoposide, and 1-(5-isoquinolinyl sulfonyl)-2-methylpiperazine (H-7). Interestingly, RA affected the ability of H-7 to induce nuclear accumulation of the p53 protein without altering its effect on elevating the steady-state level of p53, suggesting that drug-induced up-regulation and nuclear accumulation of the wild-type p53 protein are separable processes. The modulation of nuclear import of p53 protein by RA may thus represent a potential mechanism by which certain tumor cells with the wild-type p53 gene develop resistance to chemotherapeutic agents. PMID- 10364269 TI - Bivalent inhibitor of the N-end rule pathway. AB - The N-end rule relates the in vivo half-life of a protein to the identity of its N-terminal residue. Ubr1p, the recognition (E3) component of the Saccharomyces cerevisiae N-end rule pathway, contains at least two substrate-binding sites. The type 1 site is specific for N-terminal basic residues Arg, Lys, and His. The type 2 site is specific for N-terminal bulky hydrophobic residues Phe, Leu, Trp, Tyr, and Ile. Previous work has shown that dipeptides bearing either type 1 or type 2 N-terminal residues act as weak but specific inhibitors of the N-end rule pathway. We took advantage of the two-site architecture of Ubr1p to explore the feasibility of bivalent N-end rule inhibitors, whose expected higher efficacy would result from higher affinity of the cooperative (bivalent) binding to Ubr1p. The inhibitor comprised mixed tetramers of beta-galactosidase that bore both N terminal Arg (type 1 residue) and N-terminal Leu (type 2 residue) but that were resistant to proteolysis in vivo. Expression of these constructs in S. cerevisiae inhibited the N-end rule pathway much more strongly than the expression of otherwise identical beta-galactosidase tetramers whose N-terminal residues were exclusively Arg or exclusively Leu. In addition to demonstrating spatial proximity between the type 1 and type 2 substrate-binding sites of Ubr1p, these results provide a route to high affinity inhibitors of the N-end rule pathway. PMID- 10364270 TI - Parechoviruses. PMID- 10364271 TI - Variants from the diverse virus population identified at seroconversion of a clade A human immunodeficiency virus type 1-infected woman have distinct biological properties. AB - Development of effective therapeutics to prevent new infections with human immunodeficiency type 1 (HIV-1) is predicated on an understanding of the properties that provide a selective advantage to a transmitted viral population. In contrast to the homogeneous virus population that typifies early HIV-1 infection of men, the viral population in women recently infected with clade A HIV-1 is genetically diverse, based on evaluation of the envelope gene. A longitudinal study of viral envelope evolution in several women suggested that representative envelope variants detected at seroconversion had distinct biological properties that affected viral fitness. To test this hypothesis, a full-length, infectious molecular clone, Q23-17, was obtained from an infected woman 1 year following seroconversion, and chimeric viruses containing envelope genes representative of seroconversion and 27-month-postseroconversion populations were constructed. Dendritic cells (DC) could transfer infection of seroconversion variant Q23ScA, which dominated the viral population in the year following seroconversion, and the closely related 1-year isolate Q23-17 to resting peripheral blood mononuclear cells (PBMC). In contrast, resting PBMC exposed to DC pulsed with Q23ScB, which was detected infrequently in samples after seroconversion, or the 27-month chimeras were inconsistently infected. Additionally, quiescent PBMC infected with Q23ScA or Q23-17 proliferated more robustly than uninfected cells or cells infected with the other envelope chimeras in response to immobilized anti-CD3. Stimulation with tetanus toxoid led to an increased proportion of CD45RA+ cells and a decreased expression of CD28 on CD45RO+ cells in cultures of Q23-17-infected PBMC. These data demonstrate that variants from the heterogeneous seroconversion clade A HIV-1 population in a Kenyan woman have distinct biological features that may influence viral pathogenesis. PMID- 10364272 TI - Measles virus spread by cell-cell contacts: uncoupling of contact-mediated receptor (CD46) downregulation from virus uptake. AB - CD46, which serves as a receptor for measles virus (MV; strain Edmonston), is rapidly downregulated from the cell surface after contact with viral particles or infected cells. We show here that the same two CD46 complement control protein (CCP) domains responsible for primary MV attachment mediate its downregulation. Optimal downregulation efficiency was obtained with CD46 recombinants containing CCP domains 1 and 2, whereas CCP 1, alone and duplicated, induced a slight downregulation. Using persistently infected monocytic/promyelocytic U937 cells which release very small amounts of infectious virus, and uninfected HeLa cells as contact partners, we then showed that during contact the formation of CD46 containing patches and caps precedes CD46 internalization. Nevertheless, neither substances inhibiting capping nor the fusion-inhibiting peptide Z-D-Phe-L-Phe-Gly OH (FIP) blocked CD46 downregulation. Thus, CD46 downregulation can be uncoupled from fusion and subsequent virus uptake. Interestingly, in that system cell-cell contacts lead to a remarkably efficient infection of the target cells which is only partially inhibited by FIP. The finding that the contact of an infected with uninfected cells results in transfer of infectious viral material without significant (complete) fusion of the donor with the recipient cell suggests that microfusion events and/or FIP-independent mechanisms may mediate the transfer of MV infectivity from cell to cell. PMID- 10364273 TI - Characterization of an equine arteritis virus replicase mutant defective in subgenomic mRNA synthesis. AB - Equine arteritis virus (EAV) is a positive-stranded RNA virus that synthesizes a 5'- and 3'-coterminal nested set of six subgenomic mRNAs. These mRNAs all contain a common leader sequence which is derived from the 5' end of the genome. Subgenomic mRNA transcription and genome replication are directed by the viral replicase, which is expressed in the form of two polyproteins and subsequently processed into smaller nonstructural proteins (nsps). During the recent construction of an EAV infectious cDNA clone (pEAV030 [L. C. van Dinten, J. A. den Boon, A. L. M. Wassenaar, W. J. M. Spaan, and E. J. Snijder, Proc. Natl. Acad. Sci. USA 94:991-996, 1997]), a mutant cDNA clone (pEAV030F) which carries a single replicase point mutation was obtained. This substitution (Ser-2429-->Pro) is located in the nsp10 subunit and renders the EAV030F virus deficient in subgenomic mRNA synthesis. To obtain more insight into the role of nsp10 in transcription and the nature of the transcriptional defect, we have now analyzed the EAV030F mutant in considerable detail. The Ser-2429-->Pro mutation does not affect the proteolytic processing of the replicase but apparently affects the function of nsp10 in transcription. Furthermore, our study showed that EAV030F still produces subgenomic positive and negative strands, albeit at a very low level. Both subgenomic positive-strand synthesis and negative-strand synthesis are equally affected by the Ser-2429-->Pro mutation, suggesting that nsp10 plays an important role in an early step of EAV mRNA transcription. PMID- 10364274 TI - Role of the ATP-binding domain of the human papillomavirus type 11 E1 helicase in E2-dependent binding to the origin. AB - Replication of the genome of human papillomaviruses (HPV) is initiated by the recruitment of the viral E1 helicase to the origin of DNA replication by the viral E2 protein, which binds specifically to the origin. We determined, for HPV type 11 (HPV-11), that the C-terminal 296 amino acids of E1 are sufficient for interaction with the transactivation domain of E2 in the yeast two-hybrid system and in vitro. This region of E1 encompasses the ATP-binding domain. Here we have examined the role of this ATP-binding domain, and of ATP, on E2-dependent binding of E1 to the origin. Several amino acid substitutions in the phosphate-binding loop (P loop), which is implicated in binding the triphosphate moiety of ATP, abolished E2 binding, indicating that the structural integrity of this domain is essential for the interaction. The structural constraints imposed on the E1 P loop may differ between HPV-11 and bovine papillomavirus type 1 (BPV-1), since the P479S substitution that inactivates BPV-1 E1 is tolerated in the HPV-11 enzyme. Other substitutions in the E1 P loop, or in two other conserved motifs of the ATP-binding domain, were tolerated, indicating that ATP binding is not essential for interaction with E2. Nevertheless, ATP-Mg stimulated the E2 dependent binding of E1 to the origin in vitro. This stimulation was maximal at the physiological temperature (37 degrees C) and did not require ATP hydrolysis. In contrast, ATP-Mg did not stimulate the E2-dependent binding to the origin of an E1 protein containing only the C-terminal domain (353 to 649) or that of mutant E1 proteins with alterations in the DNA-binding domain. These results are discussed in light of a model in which the E1 ATP-binding domain is required for formation of the E2-binding surface and can, upon the binding of ATP, facilitate and/or stabilize the interaction of E1 with the origin. PMID- 10364275 TI - Selection for neutralization resistance of the simian/human immunodeficiency virus SHIVSF33A variant in vivo by virtue of sequence changes in the extracellular envelope glycoprotein that modify N-linked glycosylation. AB - We previously reported on the in vivo adaptation of an infectious molecular simian/human immunodeficiency virus (SHIV) clone, SHIVSF33, into a pathogenic biologic viral variant, designated SHIVSF33A. In the present study, we show that SHIVSF33A is resistant to neutralization by human immunodeficiency virus (HIV) and SHIV antisera. Multiple amino acid substitutions accumulated over time throughout the env gene of SHIVSF33A; some of them coincided with the acquisition of the neutralization resistance of the virus. Of interest are changes that resulted in the removal, repositioning, and addition of potential glycosylation sites within the V1, V2, and V3 regions of envelope gp120. To determine whether potential glycosylation changes within these principal neutralization domains of HIV type 1 formed the basis for the resistance to serum neutralization of SHIVSF33A, mutant viruses were generated on the backbone of parental SHIVSF33 and tested for their neutralization sensitivity. The mutations generated did not alter the in vitro replication kinetics or cytopathicity of the mutant viruses in T-cell lines. However, the removal of a potential glycosylation site in the V1 domain or the creation of such a site in the V3 domain did allow the virus to escape serum neutralization antibodies that recognized parental SHIVSF33. The combination of the V1 and V3 mutations conferred an additive effect on neutralization resistance over that of the single mutations. Taken together, these data suggest that (i) SHIV variants with changes in the Env SU can be selected in vivo primarily by virtue of their ability to escape neutralizing antibody recognition and (ii) carbohydrates play an important role in conferring neutralization escape, possibly by altering the structure of envelope gp120 or by shielding principal neutralization sites. PMID- 10364276 TI - Human memory cytotoxic T-lymphocyte (CTL) responses to Hantaan virus infection: identification of virus-specific and cross-reactive CD8(+) CTL epitopes on nucleocapsid protein. AB - Hantaan virus, the prototypic member of the Hantavirus genus, causes hemorrhagic fever with renal syndrome in humans. We examined the human memory T-lymphocyte responses of three donors who had previous laboratory-acquired infections with Hantaan virus. We demonstrated virus-specific responses in bulk cultures of peripheral blood mononuclear cells (PBMC) from all donors. Bulk T-cell responses were directed against either Hantaan virus nucleocapsid (N) or G1 protein, and these responses varied between donors. We established both CD4(+) and CD8(+) N specific cell lines from two donors and CD4(+) G1-specific cell lines from a third donor. All CD8(+) cytotoxic T-lymphocyte (CTL) lines recognized one of two epitopes on the nucleocapsid protein: one epitope spanning amino acids 12 to 20 and the other spanning amino acids 421 to 429. The CTL lines specific for amino acids 12 to 20 were restricted by HLA B51, and those specific for amino acids 421 to 429 were restricted by HLA A1. The N-specific CTL lines isolated from these two donors included both Hantaan virus-specific CTLs and hantavirus cross reactive CTLs. Responses to both epitopes are detectable in short-term bulk cultures of PBMC from one donor, and precursor frequency analysis confirms that CTLs specific for these epitopes are present at relatively high precursor frequencies in the peripheral T-cell pool. These data suggest that infection with Hantaan virus results in the generation of CTL to limited epitopes on the nucleocapsid protein and that infection also results in the generation of cross reactive T-cell responses to distantly related hantaviruses which cause the distinct hantavirus pulmonary syndrome. This is the first demonstration of human T-lymphocyte responses to Hantaan virus. PMID- 10364277 TI - In vitro assembly of alphavirus cores by using nucleocapsid protein expressed in Escherichia coli. AB - The production of the alphavirus virion is a multistep event requiring the assembly of the nucleocapsid core in the cytoplasm and the maturation of the glycoproteins in the endoplasmic reticulum and the Golgi apparatus. These components associate during the budding process to produce the mature virion. The nucleocapsid proteins of Sindbis virus and Ross River virus have been produced in a T7-based Escherichia coli expression system and purified. In the presence of single-stranded but not double-stranded nucleic acid, the proteins oligomerize in vitro into core-like particles which resemble the native viral nucleocapsid cores. Despite their similarities, Sindbis virus and Ross River virus capsid proteins do not form mixed core-like particles. Truncated forms of the Sindbis capsid protein were used to establish amino acid requirements for assembly. A capsid protein starting at residue 19 [CP(19-264)] was fully competent for in vitro assembly, whereas proteins with further N-terminal truncations could not support assembly. However, a capsid protein starting at residue 32 or 81 was able to incorporate into particles in the presence of CP(19-264) or could inhibit assembly if its molar ratio relative to CP(19-264) was greater than 1:1. This system provides a basis for the molecular dissection of alphavirus core assembly. PMID- 10364278 TI - Immunogenicity of a human immunodeficiency virus (HIV) polytope vaccine containing multiple HLA A2 HIV CD8(+) cytotoxic T-cell epitopes. AB - Compelling evidence now suggests that alphabeta CD8 cytotoxic T lymphocytes (CTL) have an important role in preventing human immunodeficiency virus (HIV) infection and/or slowing progression to AIDS. Here, we describe an HIV type 1 CTL polyepitope, or polytope, vaccine comprising seven contiguous minimal HLA A2 restricted CD8 CTL epitopes conjoined in a single artificial construct. Epitope specific CTL lines derived from HIV-infected individuals were able to recognize every epitope within the construct, and HLA A2-transgenic mice immunized with a recombinant virus vaccine coding for the HIV polytope also generated CTL specific for different epitopes. Each epitope in the polytope construct was therefore processed and presented, illustrating the feasibility of the polytope approach for HIV vaccine design. By simultaneously inducing CTL specific for different epitopes, an HIV polytope vaccine might generate activity against multiple challenge isolates and/or preempt the formation of CTL escape mutants. PMID- 10364279 TI - Effects of exonuclease activity and nucleotide selectivity of the herpes simplex virus DNA polymerase on the fidelity of DNA replication in vivo. AB - A mutagenesis system was developed for the in vivo study of the fidelity of DNA replication mediated by wild-type herpes simplex virus type 1 (HSV-1) strain KOS and its polymerase (Pol) mutant derivatives PAAr5, Y7, and YD12. The pHOS1 shuttle plasmid, which contained the SupF mutagenesis marker gene and the HSV oris sequence, was used for analysis of the mutation frequency and the mutation spectrum. All three Pol mutants induced significant increases in the mutation frequencies of the target gene, despite the fact that PAAr5 was previously shown to have an antimutator phenotype by the thymidine kinase mutagenesis assay (J. D. Hall, D. M. Coen, B. L. Fisher, M. Weisslitz, S. Randall, R. E. Almy, P. Gelep, and P. A. Schaffer, Virology 132:26-37, 1984; C. B. C. Hwang and J.-H. Chen, Gene 152:191-193, 1995). Altered spectra of mutated target genes induced by these three mutants were also observed. The relative frequencies of both deletion and complex mutations found in mutants induced by exonuclease-proficient Pols were significantly higher than those induced by exonuclease-deficient Pols. On the other hand, the exonuclease-deficient Pols induced significant increases in the frequency of base substitutions, which comprised predominantly G. C-to-T. A transversions, as well as mutations at additional hot spots. These results suggest that the HSV-1 DNA Pol can incorporate purine-purine or pyrimidine pyrimidine mispaired bases which may be preferentially proofread by its intrinsic exonuclease activity. Furthermore, the effects of the sequence context of the target gene and the assay method should also be considered carefully in any analysis of replication fidelity. PMID- 10364280 TI - p53-Independent and -dependent requirements for E1B-55K in adenovirus type 5 replication. AB - The adenovirus type 5 mutant dl1520 was engineered previously to be completely defective for E1B-55K functions. Recently, this mutant (also known as ONYX-015) has been suggested to replicate preferentially in p53(-) and some p53(+) tumor cell lines but to be attenuated in primary cultured cells (C. Heise, A. Sampson Johannes, A. Williams, F. McCormick, D. D. F. Hoff, and D. H. Kirn, Nat. Med. 3:639-645, 1997). It has been suggested that dl1520 might be used as a "magic bullet" that could selectively lyse tumor cells without harm to normal tissues. However, we report here that dl1520 replication is independent of p53 genotype and occurs efficiently in some primary cultured human cells, indicating that the mutant virus does not possess a tumor selectivity. Although it was not the sole host range determinant, p53 function did reduce dl1520 replication when analyzed in a cell line expressing temperature-sensitive p53 (H1299-tsp53) (K. L. Fries, W. E. Miller, and N. Raab-Traub, J. Virol. 70:8653-8659, 1996). As found earlier for other E1B-55K mutants in HeLa cells (Y. Ho, R. Galos, and J. Williams, Virology 122:109-124, 1982), dl1520 replication was temperature dependent in H1299 cells. When p53 function was restored at low temperature in H1299-tsp53 cells, it imposed a modest defect in viral DNA replication and accumulation of late viral cytoplasmic mRNA. However, in both H1299 and H1299-tsp53 cells, the defect in late viral protein synthesis appeared to be much greater than could be accounted for by the modest defects in late viral mRNA levels. We therefore propose that in addition to countering p53 function and modulating viral and cellular mRNA nuclear transport, E1B-55K also stimulates late viral mRNA translation. PMID- 10364281 TI - Genome structure and expression of the ev/J family of avian endogenous viruses. AB - We recently reported the identification of sequences in the chicken genome that show over 95% identity to the novel envelope gene of the subgroup J avian leukosis virus (S. J. Benson, B. L. Ruis, A. M. Fadly, and K. F. Conklin, J. Virol. 72:10157-10164, 1998). Based on the fact that the endogenous subgroup J related env genes were associated with long terminal repeats (LTRs), we concluded that these LTR-env sequences defined a new family of avian endogenous viruses that we designated the ev/J family. In this report, we have further characterized the content and expression of the ev/J proviruses. The data obtained indicate that there are between 6 and 11 copies of ev/J proviruses in all chicken cells examined and that these proviruses fall into six classes. Of the 18 proviruses examined, all share a high degree of sequence identity and all contain an internal deletion that removes all of the pol gene and various amounts of gag and env gene sequences. Sequencing of the gag genes, LTRs, and untranslated regions of several ev/J proviruses revealed a high level of identity between isolates, indicating that they have not undergone significant sequence variation since their introduction into the avian germ line. Although the ev/J gag gene showed a relatively weak relationship (46% identity and 61% similarity at the amino acid level) to that of the avian leukosis-sarcoma virus family, it retains several sequences of demonstrated importance for virus assembly, budding, and/or infectivity. Finally, evidence was obtained that at least some members of the ev/J family are expressed and, if translated, could encode Gag- and Env-related polypeptides. PMID- 10364282 TI - Comparative fitness of multi-dideoxynucleoside-resistant human immunodeficiency virus type 1 (HIV-1) in an In vitro competitive HIV-1 replication assay. AB - We examined whether human immunodeficiency virus type 1 (HIV-1) fitness was altered upon the acquisition of a set or subset of five mutations (A62V, V75I, F77L, F116Y, and Q151M) in the pol gene, which confers resistance to multiple dideoxynucleosides (MDR), as well as the zidovudine resistance-associated mutation T215Y, using a competitive HIV-1 replication assay in a setting of an HXB2D genetic background. Target H9 cells were exposed to a 50:50 mixture of paired infectious molecular clones, and HIV-1 in the culture supernatant was transmitted to new cultures every 7 to 10 days. The polymerase-encoding region of the virus was sequenced at various time points, and the relative proportion of the two viral populations was determined. In the absence of drugs, the comparative order for replicative fitness was HIV-162/75/77/116/151 > HIV 177/116/151 > HIV-1151 > wild-type HIV-1 (HIV-1wt) > HIV-175/77/116/151 > HIV 1151/215 > HIV-1215. In the presence of zidovudine or didanosine, the order was HIV-162/75/77/116/151 > HIV-177/116/151 > HIV-175/77/116/151 > HIV-1151 > HIV 1215. HIV-1215S(TCC), a putative intermediate infectious clone for HIV-1215, replicated comparably to HIV-1wt, while two putative intermediates for HIV-1151 [HIV-1151L(CTG) and HIV-1151K(AAG)] replicated much less efficiently than HIV-1wt and HIV-1151, suggesting that for HIV-1151 to develop, two base substitutions are likely to occur concurrently or within a short interval. These data may illustrate the molecular basis by which HIV-1151 emerges much less frequently than HIV-1215. The present data also demonstrate that several MDR HIV-1 variants are more fit than HIV-1wt in the absence of drugs and that resistance-associated mutations and drug pressure are critical variates for HIV-1 fitness. PMID- 10364283 TI - Inhibition of virion maturation by simultaneous deletion of glycoproteins E, I, and M of pseudorabies virus. AB - Glycoprotein M (gM), the product of the UL10 gene of pseudorabies virus (PrV), is one of the few nonessential glycoproteins conserved throughout the Herpesviridae. In contrast to wild-type PrV strains, the UL10 gene product of the attenuated PrV vaccine strain Bartha (PrV-Ba) is not modified by N-glycans due to a mutation in the DNA sequence encoding the consensus N-glycosylation motif. To assay function of the UL10 protein in PrV-Ba, a UL10-deletion mutant (PrV-Ba-UL10(-)) was isolated. Surprisingly, in contrast to gM-deleted wild-type PrV, PrV-Ba-UL10(-) was severely impaired in plaque formation, inducing only foci of very few infected RK13, Vero, and PSEK cells and tiny plaques on MDBK cells. Since this effect was significantly more dramatic than in wild-type PrV, additional mutations known to be present in PrV-Ba were analyzed for their contribution to this phenotype. trans-complementation of the mutated PrV-Ba UL21 or gC protein by the wild-type version had no influence on the observed phenotype. In contrast, complementation of the gE/gI deletion rescued the phenotype. The synergistic effect of deletions in gE/gI and gM on plaque size was verified by construction of a gE/I/M triple mutant derived from wild-type PrV which exhibited the same phenotype. The dramatic effect of deletion of gM on plaque size in a gE/I- virus background was mainly attributable to a function of gM, and not of the gM/gN complex, as shown by analysis of a gE/I/N triple mutant. Interestingly, despite the strong effect on plaque size, penetration was not significantly impaired. In noncomplementing cells infected with the gE/I/M triple mutant, electron microscopy showed absence of secondary envelopment in the cytoplasm but occurrence of intracytoplasmic accumulations of nucleocapsids in association with electron dense material, presumably tegument proteins. These structures were not observed after infection of cells expressing either gE/I or gM. We suggest that gE/I and gM are required for late stages in virion morphogenesis prior to final envelopment in the cytoplasm. PMID- 10364284 TI - Effects of soluble CD4 on simian immunodeficiency virus infection of CD4-positive and CD4-negative cells. AB - A soluble form of the CD4 receptor (sCD4) can either enhance or inhibit the infection of cells by simian immunodeficiency virus (SIV) and human immunodeficiency virus. We investigated the basis for these varying effects by studying the entry of three SIV isolates into CD4-positive and CD4-negative cells expressing different chemokine receptors. Infection of CD4-negative cells depended upon the viral envelope glycoproteins and upon the chemokine receptor, with CCR5 and gpr15 being more efficient than STRL33. Likewise, enhancement of infection by sCD4 was observed when CCR5- and gpr15-expressing target cells were used but not when those expressing STRL33 were used. The sCD4-mediated enhancement of virus infection of CD4-negative, CCR5-positive cells was related to the sCD4-induced increase in binding of the viral gp120 envelope glycoprotein to CCR5. Inhibitory effects of sCD4 could largely be explained by competition for virus attachment to cellular CD4 rather than other detrimental effects on virus infectivity (e.g., disruption of the envelope glycoprotein spike). Consistent with this, the sCD4-activated SIV envelope glycoprotein intermediate on the virus was long-lived. Thus, the net effect of sCD4 on SIV infectivity appears to depend upon the degree of enhancement of chemokine receptor binding and upon the efficiency of competition for cellular CD4. PMID- 10364285 TI - Combinatorial screening and intracellular antiviral activity of hairpin ribozymes directed against hepatitis B virus. AB - A combinatorial screening method has been used to identify hairpin ribozymes that inhibit hepatitis B virus (HBV) replication in transfected human hepatocellular carcinoma (HCC) cells. A hairpin ribozyme library (5 x 10(5) variants) containing a randomized substrate-binding domain was used to identify accessible target sites within 3.3 kb of full-length in vitro-transcribed HBV pregenomic RNA. Forty potential target sites were found within the HBV pregenomic RNA, and 17 sites conserved in all four subtypes of HBV were chosen for intracellular inhibition experiments. Polymerase II and III promoter expression constructs for corresponding hairpin ribozymes were generated and cotransfected into HCC cells together with a replication-competent dimer of HBV DNA. Four ribozymes inhibited HBV replication by 80, 69, 66, and 49%, respectively, while catalytically inactive mutant forms of these ribozymes affected HBV replication by 36, 28, 0, and 0%. These findings indicate that the inhibitory effects on HBV replication were largely mediated by the catalytic activity of the ribozymes. In conclusion, we have identified catalytically active RNAs by combinatorial screening that mediate intracellular antiviral effects on HBV. PMID- 10364287 TI - Induction of human papillomavirus-specific CD4(+) and CD8(+) lymphocytes by E7 pulsed autologous dendritic cells in patients with human papillomavirus type 16- and 18-positive cervical cancer. AB - Human papillomavirus (HPV) type 16 (HPV 16) and HPV type 18 (HPV 18) are implicated in the induction and progression of the majority of cervical cancers. Since the E6 and E7 oncoproteins of these viruses are expressed in these lesions, such proteins might be potential tumor-specific targets for immunotherapy. In this report, we demonstrate that recombinant, full-length E7-pulsed autologous dendritic cells (DC) can elicit a specific CD8(+) cytotoxic T-lymphocyte (CTL) response against autologous tumor target cells in three patients with HPV 16- or HPV 18-positive cervical cancer. E7-specific CTL populations expressed strong cytolytic activity against autologous tumor cells, did not lyse autologous concanavalin A-treated lymphoblasts or autologous Epstein-Barr virus-transformed lymphoblastoid cell lines (LCL), and showed low levels of cytotoxicity against natural killer cell-sensitive K562 cells. Cytotoxicity against autologous tumor cells could be significantly blocked by anti-HLA class I (W6/32) and anti CD11a/LFA-1 antibodies. Phenotypically, all CTL populations were CD3(+)/CD8(+), with variable levels of CD56 expression. CTL induced by E7-pulsed DC were also highly cytotoxic against an allogeneic HLA-A2(+) HPV 16-positive matched cell line (CaSki). In addition, we show that specific lymphoproliferative responses by autologous CD4(+) T cells can also be induced by E7-pulsed autologus DC. E7 specific CD4(+) T cells proliferated in response to E7-pulsed LCL but not unpulsed LCL, and this response could be blocked by anti-HLA class II antibody. Finally, with two-color flow cytometric analysis of intracellular cytokine expression at the single-cell level, a marked Th1-like bias (as determined by the frequency of gamma interferon- and interleukin 4-expressing cells) was observable for both CD8(+) and CD4(+) E7-specific lymphocyte populations. Taken together, these data demonstrate that full-length E7-pulsed DC can induce both E7-specific CD4(+) T-cell proliferative responses and strong CD8(+) CTL responses capable of lysing autologous naturally HPV-infected cancer cells in patients with cervical cancer. These results may have important implications for the treatment of cervical cancer patients with active or adoptive immunotherapy. PMID- 10364286 TI - Translation elongation factor 1-alpha interacts specifically with the human immunodeficiency virus type 1 Gag polyprotein. AB - Human immunodeficiency virus type 1 (HIV-1) gag-encoded proteins play key functions at almost all stages of the viral life cycle. Since these functions may require association with cellular factors, the HIV-1 matrix protein (MA) was used as bait in a yeast two-hybrid screen to identify MA-interacting proteins. MA was found to interact with elongation factor 1-alpha (EF1alpha), an essential component of the translation machinery that delivers aminoacyl-tRNA to ribosomes. EF1alpha was then shown to bind the entire HIV-1 Gag polyprotein. This interaction is mediated not only by MA, but also by the nucleocapsid domain, which provides a second, independent EF1alpha-binding site on the Gag polyprotein. EF1alpha is incorporated within HIV-1 virion membranes, where it is cleaved by the viral protease and protected from digestion by exogenously added subtilisin. The specificity of the interaction is demonstrated by the fact that EF1alpha does not bind to nonlentiviral MAs and does not associate with Moloney murine leukemia virus virions. The Gag-EF1alpha interaction appears to be mediated by RNA, in that basic residues in MA and NC are required for binding to EF1alpha, RNase disrupts the interaction, and a Gag mutant with undetectable EF1alpha-binding activity is impaired in its ability to associate with tRNA in cells. Finally, the interaction between MA and EF1alpha impairs translation in vitro, a result consistent with a previously proposed model in which inhibition of translation by the accumulation of Gag serves to release viral RNA from polysomes, permitting the RNA to be packaged into nascent virions. PMID- 10364288 TI - Identification of the RNA-binding, dimerization, and eIF4GI-binding domains of rotavirus nonstructural protein NSP3. AB - The rotavirus nonstructural protein NSP3 is a sequence-specific RNA binding protein that binds the nonpolyadenylated 3' end of the rotavirus mRNAs. NSP3 also interacts with the translation initiation factor eIF4GI and competes with the poly(A) binding protein. Deletion mutations and point mutations of NSP3 from group A rotavirus (NSP3A), expressed in Escherichia coli, indicate that the RNA binding domain lies between amino acids 4 and 149. Similar results were obtained with NSP3 from group C rotaviruses. Data also indicate that a dimer of NSP3A binds one molecule of RNA and that dimerization is necessary for strong RNA binding. The dimerization domain of NSP3 was mapped between amino acids 150 and 206 by using the yeast two-hybrid system. The eukaryotic initiation factor 4 GI subunit (eIF-4GI) binding domain of NSP3A has been mapped in the last 107 amino acids of its C terminus by using a pulldown assay and the yeast two-hybrid system. NSP3 is composed of two functional domains separated by a dimerization domain. PMID- 10364289 TI - Cell cycle- and Vpr-mediated regulation of human immunodeficiency virus type 1 expression in primary and transformed T-cell lines. AB - Viral protein R (Vpr) of human immunodeficiency virus type 1 (HIV-1) transiently arrests cells in the G2 phase of the cell cycle and is a weak transcriptional transactivator. We found that Vpr increased HIV-1 long terminal repeat (LTR) activity in all cells examined but, when expressed at high levels, decreased HIV 1 LTR expression due to cytotoxic effects. Moreover, Vpr-mediated enhancement of HIV-1 LTR-driven transcription was observed in cycling primary human CD4(+) T cells but not in terminally differentiated, noncycling primary human macrophages. In single-round infection experiments using primary human CD4(+) T cells, proviral clones expressing either wild-type Vpr or Vpr mutants that retained the ability to cause a G2 arrest replicated to higher levels than proviruses lacking Vpr or expressing mutants of Vpr that did not cause an arrest. In support of the hypothesis that enhancement of HIV-1 LTR transcription by Vpr is an indirect effect of the ability of Vpr to delay cells in G2, counterflow centrifugal elutriation of cells into different phases of the cell cycle demonstrated that HIV-1 LTR expression was highest in G2. Finally, the ability of Vpr to upregulate viral transcription was dependent on a minimal promoter containing a functional TATA box and an enhancer. PMID- 10364291 TI - Isolation of recombinant adeno-associated virus vector-cellular DNA junctions from mouse liver. AB - Recombinant adeno-associated virus (rAAV) vectors allow for sustained expression of transgene products from mouse liver following a single portal vein administration. Here a rAAV vector expressing human coagulation factor F.IX (hF.IX), AAV-EF1alpha-F.IX (hF.IX expression was controlled by the human elongation factor 1alpha [EF1alpha] enhancer-promoter) was injected into mice via the portal vein or tail vein, or directly into the liver parenchyma, and the forms of rAAV vector DNA extracted from the liver were analyzed. Southern blot analyses suggested that rAAV vector integrated into the host genome, forming mainly head-to-tail concatemers with occasional deletions of the inverted terminal repeats (ITRs) and their flanking sequences. To further confirm vector integration, we developed a shuttle vector system and isolated and sequenced rAAV vector-cellular DNA junctions from transduced mouse livers. Analysis of 18 junctions revealed various rearrangements, including ITR deletions and amplifications of the vector and cellular DNA sequences. The breakpoints of the vector were mostly located within the ITRs, and cellular DNA sequences were recombined with the vector genome in a nonhomologous manner. Two rAAV-targeted DNA sequences were identified as the mouse rRNA gene and the alpha1 collagen gene. These observations serve as direct evidence of rAAV integration into the host genome of mouse liver and allow us to begin to elucidate the mechanisms involved in rAAV integration into tissues in vivo. PMID- 10364290 TI - Identification of a cytoplasmic targeting/retention signal in a retroviral Gag polyprotein. AB - Retroviral capsid assembly can occur by either of two distinct morphogenic processes: in type C viruses, the capsid assembles and buds at the plasma membrane, while in type B and D viruses, the capsid assembles within the cytoplasm and is then transported to the plasma membrane for budding. We have previously reported that a single-amino-acid substitution of a tryptophan for an arginine in the matrix protein (MA) of Mason-Pfizer monkey virus (MPMV) converts its capsid assembly from that of a type D retrovirus to that of the type C viruses (S. S. Rhee and E. Hunter, Cell 63:77-86, 1990). Here we identify a region of 18 amino acids within the MA of MPMV that is responsible for type D specific morphogenesis. Insertion of these 18 amino acids into the MA of type C Moloney murine leukemia virus causes it to assemble an immature capsid in the cytoplasm. Furthermore, fusion of the MPMV MA to the green fluorescent protein resulted in altered intracellular targeting and a punctate accumulation of the fusion protein in the cytoplasm. These 18 amino acids, which are necessary and sufficient to target retroviral Gag polyproteins to defined sites in the cytoplasm, appear to define a novel mammalian cytoplasmic targeting/retention signal. PMID- 10364292 TI - Human and rodent transcription elongation factor P-TEFb: interactions with human immunodeficiency virus type 1 tat and carboxy-terminal domain substrate. AB - The human immunodeficiency virus type 1 transcriptional regulator Tat increases the efficiency of elongation, and complexes containing the cellular kinase CDK9 have been implicated in this process. CDK9 is part of the Tat-associated kinase TAK and of the elongation factor P-TEFb (positive transcription elongation factor b), which consists minimally of CDK9 and cyclin T. TAK and P-TEFb are both able to phosphorylate the carboxy-terminal domain (CTD) of RNA polymerase II, but their relationships to one another and to the stimulation of elongation by Tat are not well characterized. Here we demonstrate that human cyclin T1 (but not cyclin T2) interacts with the activation domain of Tat and is a component of TAK as well as of P-TEFb. Rodent (mouse and Chinese hamster) cyclin T1 is defective in Tat binding and transactivation, but hamster CDK9 interacts with human cyclin T1 to give active TAK in hybrid cells containing human chromosome 12. Although TAK is phosphorylated on both serine and threonine residues, it specifically phosphorylates serine 5 in the CTD heptamer. TAK is found in the nuclear and cytoplasmic fractions of human cells as a large complex (approximately 950 kDa). Magnesium or zinc ions are required for the association of Tat with the kinase. We suggest a model in which Tat first interacts with P-TEFb to form the TAK complex that engages with TAR RNA and the elongating transcription complex, resulting in hyperphosphorylation of the CTD on serine 5 residues. PMID- 10364294 TI - Use of major histocompatibility complex class I/peptide/beta2M tetramers to quantitate CD8(+) cytotoxic T lymphocytes specific for dominant and nondominant viral epitopes in simian-human immunodeficiency virus-infected rhesus monkeys. AB - To evaluate the impact of the diversity of antigen recognition by T lymphocytes on disease pathogenesis, we must be able to identify and analyze simultaneously cytotoxic T-lymphocyte (CTL) responses specific for multiple viral epitopes. Many of the studies of the role of CD8(+) CTLs in AIDS pathogenesis have been done with simian immunodeficiency virus (SIV)- and simian-human immunodeficiency virus (SHIV)-infected rhesus monkeys. These studies have frequently made use of the well-defined SIV Gag CTL epitope p11C,C-M presented to CTL by the HLA-A homologue molecule Mamu-A*01. In the present study we identified and fine mapped two novel Mamu-A*01-restricted CTL epitopes: the SIVmac Pol-derived epitope p68A (STPPLVRLV) and the human immunodeficiency virus type 1 (HIV-1) Env-derived p41A epitope (YAPPISGQI). The frequency of CD8(+) CTLs specific for the p11C,C-M, p68A, and p41A epitopes was quantitated in the same animals with a panel of tetrameric Mamu-A*01/peptide/beta2m complexes. All SHIV-infected Mamu-A*01(+) rhesus monkeys tested had a high frequency of SIVmac Gag-specific CTLs to the p11C,C-M epitope. In contrast, only a fraction of the monkeys tested had detectable CTLs specific for the SIVmac Pol p68A and HIV-1 Env p41A epitopes, and these responses were detected at very low frequencies. Thus, the p11C,C-M specific CD8(+) CTL response is dominant and the p41A- and p68A-specific CD8(+) CTL responses are nondominant. These results indicate that CD8(+) CTL responses to dominant CTL epitopes can be readily quantitated with the tetramer technology; however, CD8(+) CTL responses to nondominant epitopes, due to the low frequency of these epitope-specific cells, may be difficult to detect and quantitate by this approach. PMID- 10364293 TI - Elimination of duck hepatitis B virus RNA-containing capsids in duck interferon alpha-treated hepatocytes. AB - Evidence is presented that the previously cloned type I duck interferon (DuIFN) cDNA encodes a homologue of mammalian interferon-alpha (IFN-alpha). Recombinant DuIFN-alpha was used to study the inhibition of duck hepatitis B virus (DHBV) replication in primary hepatocytes in order to determine the IFN-sensitive steps of the virus replication cycle. IFN-treated cells accumulated two- to threefold lower amounts of viral RNA transcripts early during infection, when IFN was added before virus. This reduction was not due to inhibition of virus entry since initial covalently closed circular DNA levels were not decreased in IFN-treated cells. Interestingly, the inhibitory effect of IFN on viral RNA levels was not observed in cells infected with a mutant DHBV that fails to synthesize core protein, suggesting that an uncharacterized core protein-mediated enhancing effect is blocked by IFN. When IFN was added at 4 days postinfection, encapsidated viral RNA pregenomes disappeared from infected cells within 3 days. This depletion was not simply due to conversion of pregenomes to DNA since depletion was not blocked by phosphonoformic acid, an inhibitor of the viral reverse transcriptase. The intracellular concentration of intact nucleocapsids was reduced, suggesting that in the presence of IFN pregenome-containing capsids were selectively depleted in hepatocytes. Thus, two steps in DHBV replication that involve the viral core protein were inhibited by DuIFN-alpha. PMID- 10364295 TI - Replication, integration, and packaging of plasmid DNA following cotransfection with baculovirus viral DNA. AB - Infection-dependent replication assays have been used to identify numerous putative origins of baculovirus replication. However, plasmid DNA, when cotransfected into insect cells with Autographa californica multinucleocapsid nucleopolyhedrovirus (AcMNPV) DNA, replicates independently of any viral sequence in cis (11). Cotransfection of transfer plasmids and baculovirus DNA is a common procedure used in generating recombinant viruses and in measuring the level of gene expression in transient-expression assays. We have examined the fate of a series of vector plasmids in cotransfection experiments. The data reveal that these plasmids replicate following cotransfection and the replication of plasmid DNA is not due to acquisition of viral putative origin sequences. The conformation of plasmid DNA replicating in the cotransfected cells was analyzed and found to exist as high-molecular-weight concatemers. Ten to 25% of the replicated plasmid DNA was integrated into multiple locations on the viral genome and was present in progeny virions following serial passage. Sequence analysis of plasmid-viral DNA junction sites revealed no homologous or conserved sequences in the proximity of the integration sites, suggesting that nonhomologous recombination was involved during the integration process. These data suggest that while a rolling-circle mechanism could be used for baculovirus DNA replication, recombination may also be involved in this process. Plasmid integration may generate large deletions of the viral genome, suggesting that the process of DNA replication in baculovirus may be prone to generation of defective genomes. PMID- 10364296 TI - Identification of replication specificity determinants in two strains of tomato leaf curl virus from New Delhi. AB - We used two strains of tomato leaf curl virus from New Delhi to investigate specificity in replication of their cognate genomes. The strains share 94% sequence identity and are referred to as severe and mild on the basis of symptoms on tomato and tobacco. Replication assays in tobacco protoplasts and plants showed that a single amino acid change, Asn10 to Asp in the N terminus of Rep protein, determines specificity for replication of the two strains based upon its interaction with the origin of replication (ori) sequences. The change of Asp10 to Asn in Rep protein of the mild strain coupled with point mutations at the 3rd and 10th nucleotides of the 13-mer binding site altered its replication ability, resulting in increased levels of virus accumulation. Similarly, changing Asn10 to Asp in Rep protein of the severe strain impaired replication of the virus and altered its severe phenotype in plants. Site-directed mutations made in ori and Asn10 of Rep protein suggested that Asn10 recognizes the third base pair of the putative binding site sequence GGTGTCGGAGTC in the severe strain. PMID- 10364297 TI - Amelioration of retroviral vector silencing in locus control region beta-globin transgenic mice and transduced F9 embryonic cells. AB - Retroviral vectors are transcriptionally silenced in hematopoietic stem cells, and this phenomenon must be overcome for effective gene therapy of blood diseases. The murine stem cell virus (MSCV) vector completely silences beta globin reporter genes regulated by locus control region (LCR) elements 5'HS2 to 5'HS4 in seven of eight transgenic mice. Here, we show that no single known MSCV silencer element is sufficient for complete LCR beta-globin transgene silencing. However, partial silencing of high-copy transgenes is conveyed by the MSCV direct repeat and promoter elements. The CpG methylation pattern of silenced and expressed MSCV promoter transgenes is virtually identical, demonstrating that silencing does not absolutely correlate with methylation status. Combined mutations in all four MSCV silencer elements leads to expression of beta-globin in 6 of 10 transgenic mice. The same mutations incorporated into the HSC1 retrovirus vector direct neo gene expression in 71% of transduced F9 embryonic carcinoma cells. These studies demonstrate that combined mutation of four retroviral silencer elements relieves complete silencing in most transgenic mice and transduced F9 cells and suggests that novel silencer elements remain. Enhanced expression of the HSC1 vector in primitive stem cells is well suited for blood gene therapy applications. PMID- 10364298 TI - Sequence variations in human immunodeficiency virus type 1 Nef are associated with different stages of disease. AB - nef alleles derived from a large number of individuals infected with human immunodeficiency virus type 1 (HIV-1) were analyzed to investigate the frequency of disrupted nef genes and to elucidate whether specific amino acid substitutions in Nef are associated with different stages of disease. We confirm that deletions or gross abnormalities in nef are rarely present. However, a comparison of Nef consensus sequences derived from 41 long-term nonprogressors and from 50 individuals with progressive HIV-1 infection revealed that specific variations are associated with different stages of infection. Five amino acid variations in Nef (T15, N51, H102, L170, and E182) were more frequently observed among nonprogressors, while nine features (an additional N-terminal PxxP motif, A15, R39, T51, T157, C163, N169, Q170, and M182) were more frequently found in progressors. Strong correlations between the frequency of these variations in Nef and both the CD4(+)-cell count and the viral load were observed. Moreover, analysis of sequential samples obtained from two progressors revealed that several variations in Nef, which were more commonly observed in patients with low CD4(+)-T-cell counts, were detected only during or after progression to immunodeficiency. Our results indicate that sequence variations in Nef are associated with different stages of HIV-1 infection and suggest a link between nef gene function and the immune status of the infected individual. PMID- 10364299 TI - Lack of viral escape and defective in vivo activation of human immunodeficiency virus type 1-specific cytotoxic T lymphocytes in rapidly progressive infection. AB - Human immunodeficiency virus type 1 (HIV-1)-specific immune responses over the course of rapidly progressive infection are not well defined. Detailed longitudinal analyses of neutralizing antibodies, lymphocyte proliferation, in vivo-activated and memory cytotoxic T-lymphocyte (CTL) responses, and viral sequence variation were performed on a patient who presented with acute HIV-1 infection, developed an AIDS-defining illness 13 months later, and died 45 months after presentation. Neutralizing-antibody responses remained weak throughout, and no HIV-1-specific lymphocyte proliferative responses were seen even early in the disease course. Strong in vivo-activated CTL directed against Env and Pol epitopes were present at the time of the initial drop in viremia but were quickly lost. Memory CTL against Env and Pol epitopes were detected throughout the course of infection; however, these CTL were not activated in vivo. Despite an initially narrow CTL response, new epitopes were not targeted as the disease progressed. Viral sequencing showed the emergence of variants within the two targeted CTL epitopes; however, viral variants within the immunodominant Env epitope were well recognized by CTL, and there was no evidence of viral escape from immune system detection within this epitope. These data demonstrate a narrowly directed, static CTL response in a patient with rapidly progressive disease. We also show that disease progression can occur in the presence of persistent memory CTL recognition of autologous epitopes and in the absence of detectable escape from CTL responses, consistent with an in vivo defect in activation of CTL. PMID- 10364300 TI - Role of naturally occurring basic amino acid substitutions in the human immunodeficiency virus type 1 subtype E envelope V3 loop on viral coreceptor usage and cell tropism. AB - To assess the role of naturally occurring basic amino acid substitutions in the V3 loop of human immunodeficiency virus type 1 (HIV-1) subtype E on viral coreceptor usage and cell tropism, we have constructed a panel of chimeric viruses with mutant V3 loops of HIV-1 subtype E in the genetic background of HIV 1LAI. The arginine substitutions naturally occurring at positions 8, 11, and 18 of the V3 loop in an HIV-1 subtype E X4 strain were systematically introduced into that of an R5 strain to generate a series of V3 loop mutant chimera. These chimeric viruses were employed in virus infectivity assays using HOS-CD4 cells expressing either CCR5 or CXCR4, peripheral blood mononuclear cells, T-cell lines, or macrophages. The arginine substitution at position 11 of the V3 loop uniformly caused the loss of infectivity in HOS-CD4-CCR5 cells, indicating that position 11 is critical for utilization of CCR5. CXCR4 usage was conferred by a minimum of two arginine substitutions, regardless of combination, whereas arginine substitutions at position 8 and 11 were required for T-cell line tropism. Nonetheless, macrophage tropism was not conferred by the V3 loop of subtype E R5 strain per se. We found that the specific combinations of amino acid changes in HIV-1 subtype E env V3 loop are critical for determining viral coreceptor usage and cell tropism. However, the ability to infect HOS-CD4 cells through either CXCR4 or CCR5 is not necessarily correlated with T-cell or macrophage tropism, suggesting that cellular tropism is not dictated solely by viral coreceptor utilization. PMID- 10364301 TI - The anamnestic neutralizing antibody response is critical for protection of mice from challenge following vaccination with a plasmid encoding the Japanese encephalitis virus premembrane and envelope genes. AB - For Japanese encephalitis (JE), we previously reported that recombinant vaccine induced protection from disease does not prevent challenge virus replication in mice. Moreover, DNA vaccines for JE can provide protection from high challenge doses in the absence of detectable prechallenge neutralizing antibodies. In the present study, we evaluated the role of postchallenge immune responses in determining the outcome of JE virus infection, using mice immunized with a plasmid, pcDNA3JEME, encoding the JE virus premembrane (prM) and envelope (E) coding regions. In the first experiment, 10 mice were vaccinated once (five animals) or twice (remainder) with 100 micrograms of pcDNA3JEME. All of these mice showed low (6 of 10) or undetectable (4 of 10) levels of neutralizing antibodies. Interestingly, eight of these animals showed a rapid rise in neutralizing antibody following challenge with 10,000 50% lethal doses of JE virus and survived for 21 days, whereas only one of the two remaining animals survived. No unimmunized animals exhibited a rise of neutralizing antibody or survived challenge. Levels of JE virus-specific immunoglobulin M class antibodies were elevated following challenge in half of the unimmunized mice and in the single pcDNA3JEME-immunized mouse that died. In the second experiment, JE virus specific primary cytotoxic T-lymphocyte (CTL) activity was detected in BALB/c mice immunized once with 100 micrograms of pcDNA3JEME 4 days after challenge, indicating a strong postchallenge recall of CTLs. In the third experiment, evaluation of induction of CTLs and antibody activity by plasmids containing portions of the prM/E cassette demonstrated that induction of CTL responses alone were not sufficient to prevent death. Finally, we showed that antibody obtained from pcDNA3JEME-immunized mice 4 days following challenge could partially protect recipient mice from lethal challenge. Taken together, these results indicate that neutralizing antibody produced following challenge provides the critical protective component in pcDNA3JEME-vaccinated mice. PMID- 10364302 TI - Core-binding factor influences the disease specificity of Moloney murine leukemia virus. AB - The core site in the Moloney murine leukemia virus (Moloney MLV) enhancer was previously shown to be an important determinant of the T-cell disease specificity of the virus. Mutation of the core site resulted in a significant shift in disease specificity of the Moloney virus from T-cell leukemia to erythroleukemia. We and others have since determined that a protein that binds the core site, one of the core-binding factors (CBF) is highly expressed in thymus and is essential for hematopoiesis. Here we test the hypothesis that CBF plays a critical role in mediating pathogenesis of Moloney MLV in vivo. We measured the affinity of CBF for most core sites found in MLV enhancers, introduced sites with different affinities for CBF into the Moloney MLV genome, and determined the effects of these sites on viral pathogenesis. We found a correlation between CBF affinity and the latent period of disease onset, in that Moloney MLVs with high-affinity CBF binding sites induced leukemia following a shorter latent period than viruses with lower-affinity sites. The T-cell disease specificity of Moloney MLV also appeared to correlate with the affinity of CBF for its binding site. The data support a role for CBF in determining the pathogenic properties of Moloney MLV. PMID- 10364303 TI - Matrix metalloproteinase 9 expression is induced by Epstein-Barr virus latent membrane protein 1 C-terminal activation regions 1 and 2. AB - Nasopharyngeal carcinoma (NPC), which is closely associated with the Epstein-Barr virus (EBV), is a highly metastatic malignant tumor. An important activity in tumor invasion and metastasis is that of the 92-kDa type IV collagenase or gelatinase, matrix metalloproteinase 9 (MMP-9), which mediates the degradation of the basement membrane and extracellular matrix. The expression of MMP-9 has been shown to be enhanced by the EBV oncoprotein, latent membrane protein 1 (LMP-1). LMP-1, which is expressed in NPC, has two essential signaling domains within the carboxy terminus, termed C-terminal activation regions 1 (CTAR-1) and CTAR-2. This study reveals that either signaling domain can activate the MMP-9 promoter and induce MMP-9 activity; however, LMP-1 deletion mutants lacking either CTAR-1 or CTAR-2 had a decreased ability to induce MMP-9 expression. The deletion of both activation regions completely abolished the induction of MMP-9 activity, while the cotransfection of both the CTAR-1 and CTAR-2 deletion mutants restored MMP-9 activity to levels produced by wild-type LMP-1. The NF-kappaB and activator protein 1 (AP-1) binding sites in the MMP-9 promoter were essential for the activation of MMP-9 gene expression by both CTAR-1 and CTAR-2. The induction of MMP-9 expression by LMP-1 and both CTAR-1 and CTAR-2 mutants was blocked by the overexpression of IkappaB. The tumor necrosis factor receptor-associated factor (TRAF) pathway also contributed to the activation of the MMP-9 promoter as shown by the use of TRAF-2 and TRAF-3 dominant-negative constructs. These data indicate that the activation of both the NF-kappaB and AP-1 pathways by LMP-1, CTAR-1, and CTAR-2 is necessary for the activation of MMP-9 expression. In NPC, LMP-1 may contribute to invasiveness and metastasis through the induction of MMP-9 transcription and enzymatic activity. PMID- 10364304 TI - Identification of the immediate-early transcripts of Kaposi's sarcoma-associated herpesvirus. AB - In the immediate-early phase of reactivation or primary infection, herpesviruses express a small number of genes without requiring prior viral protein synthesis. Immediate-early genes usually encode regulatory proteins critical for the viral life cycle. Kaposi's sarcoma-associated herpesvirus (KSHV) gene transcription in the immediate-early stage of viral reactivation was examined by using a chemical induction combined with a gene expression screening method. RNA transcripts from at least four KSHV genomic loci accumulate when latently infected B-lymphoma cells are induced for reactivation in the presence of an inhibitor of protein synthesis (cycloheximide) and thus represent immediate-early class transcripts. Among them, a 3.6-kb mRNA encodes three putative open reading frames (ORFs), namely, ORF50, K8, and K8.2. ORF50 is a homologue of Rta, a transcription activator encoded by Epstein-Barr virus (EBV). The K8 gene codes for a 237-amino acid protein with a basic-leucine zipper domain near its C terminus and an acidic domain near its N terminus and which closely resembles the ZEBRA protein of EBV and Jun/Fos family proteins. Other immediate-early mRNAs of KSHV include a 1. 7 kb mRNA encoding ORF45, a 2.0-kb mRNA encoding ORF K4.2, and a 4. 5-kb mRNA. Functional roles of products of these KSHV immediate-early transcripts remain to be studied. PMID- 10364305 TI - The versatility of paramyxovirus RNA polymerase stuttering. AB - Paramyxoviruses cotranscriptionally edit their P gene mRNAs by expanding the number of Gs of a conserved AnGn run. Different viruses insert different distributions of guanylates, e.g., Sendai virus inserts a single G, whereas parainfluenza virus type 3 inserts one to six Gs. The sequences conserved at the editing site, as well as the experimental evidence, suggest that the insertions occur by a stuttering process, i.e., by pseudotemplated transcription. The number of times the polymerase "stutters" at the editing site before continuing strictly templated elongation is directed by a cis-acting sequence found upstream of the insertions. We have examined the stuttering process during natural virus infections by constructing recombinant Sendai viruses with mutations in their cis acting sequences. We found that the template stutter site is precisely determined (C1052) and that a relatively short region (approximately 6 nucleotides) just upstream of the AnGn run can modulate the overall frequency of mRNA editing as well as the distribution of the nucleotide insertions. The positions more proximal to the 5' AnGn run are the most important in this respect. We also provide evidence that the stability of the mRNA/template hybrid plays a determining role in the overall frequency and range of mRNA editing. When the template U run is extended all the way to the stutter site, adenylates rather than guanylates are added at the editing site and their distribution begins to resemble the polyadenylation associated with mRNA 3' end formation by the viral polymerase. Our data suggest how paramyxovirus mRNA editing and polyadenylation are related mechanistically and how editing sites may have evolved from poly(A) termination sites or vice versa. PMID- 10364306 TI - Shift of clinical human immunodeficiency virus type 1 isolates from X4 to R5 and prevention of emergence of the syncytium-inducing phenotype by blockade of CXCR4. AB - The emergence of X4 human immunodeficiency virus type 1 (HIV-1) strains in HIV-1 infected individuals has been associated with CD4(+) T-cell depletion, HIV mediated CD8(+) cell apoptosis, and an impaired humoral response. The bicyclam AMD3100, a selective antagonist of CXCR4, selected for the outgrowth of R5 virus after cultivation of mixtures of the laboratory-adapted R5 (BaL) and X4 (NL4-3) HIV strains in the presence of the compound. The addition of AMD3100 to peripheral blood mononuclear cells infected with X4 or R5X4 clinical HIV isolates displaying the syncytium-inducing phenotype resulted in a complete suppression of X4 variants and a concomitant genotypic change in the V2 and V3 loops of the envelope gp120 glycoprotein. The recovered viruses corresponded genotypically and phenotypically to R5 variants in that they could no longer use CXCR4 as coreceptor or induce syncytium formation in MT-2 cells. Furthermore, the phenotype and genotype of a cloned R5 HIV-1 virus converted to those of the R5X4 virus after prolonged culture in lymphoid cells. However, these changes did not occur when the infected cells were cultured in the presence of AMD3100, despite low levels of virus replication. Our findings indicate that selective blockade of the CXCR4 receptor prevents the switch from the less pathogenic R5 HIV to the more pathogenic X4 HIV strains, a process that heralds the onset of AIDS. In this article, we show that it could be possible to redirect the evolution of HIV so as to impede the emergence of X4 strains or to change the phenotype of already existing X4 isolates to R5. PMID- 10364307 TI - Isolation and characterization of a hantavirus from Lemmus sibiricus: evidence for host switch during hantavirus evolution. AB - A novel hantavirus, first detected in Siberian lemmings (Lemmus sibiricus) collected near the Topografov River in the Taymyr Peninsula, Siberia (A. Plyusnin et al., Lancet 347:1835-1836, 1996), was isolated in Vero E6 cells and in laboratory-bred Norwegian lemmings (Lemmus lemmus). The virus, named Topografov virus (TOP), was most closely related to Khabarovsk virus (KBR) and Puumala viruses (PUU). In a cross focus reduction neutralization test, anti-TOP Lemmus antisera showed titers at least fourfold higher with TOP than with other hantaviruses; however, a rabbit anti-KBR antiserum neutralized TOP and KBR at the same titer. The TOP M segment showed 77% nucleotide and 88% amino acid identity with KBR and 76% nucleotide and 82% amino acid identity with PUU. However, the homology between TOP and the KBR S segment was disproportionately higher: 88% at the nucleotide level and 96% at the amino acid level. The 3' noncoding regions of KBR and the TOP S and M segments were alignable except for 113- and 58-nucleotide deletions in KBR. The phylogenetic relationships of TOP, KBR, and PUU and their respective rodent carriers suggest that an exceptional host switch took place during the evolution of these viruses; while TOP and KBR are monophyletic, the respective rodent host species are only distantly related. PMID- 10364308 TI - ICP22 and the UL13 protein kinase are both required for herpes simplex virus induced modification of the large subunit of RNA polymerase II. AB - Herpes simplex virus type 1 (HSV-1) infection alters the phosphorylation of the large subunit of RNA polymerase II (RNAP II), resulting in the depletion of the hypophosphorylated and hyperphosphorylated forms of this polypeptide (known as IIa and IIo, respectively) and induction of a novel, alternatively phosphorylated form (designated IIi). We previously showed that the HSV-1 immediate-early protein ICP22 is involved in this phenomenon, since induction of IIi and depletion of IIa are deficient in cells infected with 22/n199, an HSV-1 ICP22 nonsense mutant (S. A. Rice, M. C. Long, V. Lam, P. A. Schaffer, and C. A. Spencer, J. Virol. 69:5550-5559, 1995). However, depletion of IIo still occurs in 22/n199-infected cells. This suggests either that another viral gene product affects the RNAP II large subunit or that the truncated ICP22 polypeptide encoded by 22/n199 retains residual activity which leads to IIo depletion. To distinguish between these possibilities, we engineered an HSV-1 ICP22 null mutant, d22-lacZ, and compared it to 22/n199. The two mutants are indistinguishable in their effects on the RNAP II large subunit, suggesting that an additional viral gene product is involved in altering RNAP II. Two candidates are UL13, a protein kinase which has been implicated in ICP22 phosphorylation, and the virion host shutoff (Vhs) factor, the expression of which is positively regulated by ICP22 and UL13. To test whether UL13 is involved, a UL13-deficient viral mutant, d13 lacZ, was engineered. This mutant was defective in IIi induction and IIa depletion, displaying a phenotype very similar to that of d22-lacZ. In contrast, a Vhs mutant had effects that were indistinguishable from wild-type HSV-1. Therefore, UL13 but not the Vhs function plays a role in modifying the RNAP II large subunit. To study the potential role of UL13 in viral transcription, we carried out nuclear run-on transcription analyses in infected human embryonic lung cells. Infections with either UL13 or ICP22 mutants led to significantly reduced amounts of viral genome transcription at late times after infection. Together, our results suggest that ICP22 and UL13 are involved in a common pathway that alters RNAP II phosphorylation and that in some cell lines this change promotes viral late transcription. PMID- 10364309 TI - Mapping of functional elements in the stem-anchor region of tick-borne encephalitis virus envelope protein E. AB - Envelope protein E of the flavivirus tick-borne encephalitis virus mediates membrane fusion, and the structure of the N-terminal 80% of this 496-amino-acid long protein has been shown to differ significantly from that of other viral fusion proteins. The structure of the carboxy-terminal 20%, the stem-anchor region, is not known. It contains sequences that are important for membrane anchoring, interactions with prM (the precursor of membrane protein M) during virion assembly, and low-pH-induced structural changes associated with the fusion process. To identify specific functional elements in this region, a series of C terminal deletion mutants were constructed and the properties of the resulting truncated recombinant E proteins were examined. Full-length E proteins and proteins lacking the second of two predicted transmembrane segments were secreted in a particulate form when coexpressed with prM, whereas deletion of both segments resulted in the secretion of soluble homodimeric E proteins. Sites located within a predicted alpha-helical region of the stem (amino acids 431 to 449) and the first membrane-spanning region (amino acids 450 to 472) were found to be important for the stability of the prM-E heterodimer but not essential for prM-mediated intracellular transport and secretion of soluble E proteins. A separate site in the stem, also corresponding to a predicted alpha-helix (amino acids 401 to 413), was essential for the conversion of soluble protein E dimers to a homotrimeric form upon low-pH treatment, a process resembling the transition to the fusogenic state in whole virions. This functional mapping will aid in the understanding of the molecular mechanisms of membrane fusion and virus assembly. PMID- 10364310 TI - Optimal replication activity of vesicular stomatitis virus RNA polymerase requires phosphorylation of a residue(s) at carboxy-terminal domain II of its accessory subunit, phosphoprotein P. AB - The phosphoprotein, P, of vesicular stomatitis virus (VSV) is a key subunit of the viral RNA-dependent RNA polymerase complex. The protein is phosphorylated at multiple sites in two different domains. We recently showed that specific serine and threonine residues within the amino-terminal acidic domain I of P protein must be phosphorylated for in vivo transcription activity, but not for replication activity, of the polymerase complex. To examine the role of phosphorylation of the carboxy-terminal domain II residues of the P protein in transcription and replication, we have used a panel of mutant P proteins in which the phosphate acceptor sites (Ser-226, Ser-227, and Ser-233) were altered to alanines either individually or in various combinations. Analyses of the mutant proteins for their ability to support replication of a VSV minigenomic RNA suggest that phosphorylation of either Ser-226 or Ser-227 is necessary for optimal replication activity of the protein. The mutant protein (P226/227) in which both of these residues were altered to alanines was only about 8% active in replication compared to the wild-type (wt) protein. Substitution of alanine for Ser-233 did not have any adverse effect on replication activity of the protein. In contrast, all the mutant proteins showed activities similar to that of the wt protein in transcription. These results indicate that phosphorylation of the carboxy-terminal domain II residues of P protein are required for optimal replication activity but not for transcription activity. Furthermore, substitution of glutamic acid residues for Ser-226 and Ser-227 resulted in a protein that was only 14% active in replication but almost fully active in transcription. Taken together, these results, along with our earlier studies, suggest that phosphorylation of residues at two different domains in the P protein regulates its activity in transcription and replication of the VSV genome. PMID- 10364311 TI - Failure To cleave murine leukemia virus envelope protein does not preclude its incorporation in virions and productive virus-receptor interaction. AB - It is thought that complete cleavage of retroviral envelope protein into mature surface protein (SU) and transmembrane protein (TM) is critical for its assembly into virions and the formation of infectious virus particles. Here we report the identification of highly infectious, cleavage-deficient envelope mutant proteins. Substitution of aspartate for lysine 104, arginines 124 and 126, or arginines 223 and 225 strongly suppressed cleavage of the envelope precursor and yet allowed efficient incorporation of precursor molecules as the predominant species in virions that were almost as infectious as the wild-type virus. These results indicate that cleavage of the envelope precursor into mature SU and TM is not necessary for assembly into virions. Moreover, they call into question how many mature envelope protein subunits are required to complete virus entry, suggesting that a very few molecules suffice. The failure of host cell proteases to cleave these mutant proteins, whose substitutions are distal to the actual site of cleavage, suggests that the envelope precursor is misfolded, sequestering the cleavage site. In agreement with this, all cleavage mutant proteins exhibited significant losses of receptor binding, suggesting that these residues play roles in proper envelope protein folding. We also identified a charged residue, arginine 102, whose substitution suppressed envelope cleavage and allowed precursor incorporation but resulted in virions that were virtually noninfectious and that exhibited the greatest reduction in receptor binding. Placement of these cleavage mutations into envelope proteins of targeted retroviral vectors for human gene therapy may prevent loss of the modified surface proteins from virions, improving their infectivity and storage hardiness. PMID- 10364312 TI - gamma delta+ T cells regulate major histocompatibility complex class II(IA and IE)-dependent susceptibility to coxsackievirus B3-induced autoimmune myocarditis. AB - Coxsackievirus B3 (CVB3) infection induces myocardial inflammation and myocyte necrosis in some, but not all, strains of mice. C57BL/6 mice, which inherently lack major histocompatibility complex (MHC) class II IE antigen, develop minimal cardiac lesions despite high levels of virus in the heart. The present experiments evaluate the relative roles of class II IA and IE expression on myocarditis susceptibility in four transgenic C57BL/6 mouse strains differing in MHC class II antigen expression. Animals lacking MHC class II IE antigen (C57BL/6 [IA+ IE-] and ABo [IA- IE-]) developed minimal cardiac lesions subsequent to infection despite high concentrations of virus in the heart. In contrast, strains expressing IE (ABo Ealpha [IA- IE+] and Bl.Tg.Ealpha [IA+ IE+]) had substantial cardiac injury. Myocarditis susceptibility correlated to a Th1 (gamma interferon positive) cell response in the spleen, while disease resistance correlated to a preferential Th2 (interleukin-4-positive) phenotype. Vgamma/Vdelta analysis indicates that distinct subpopulations of gamma delta+ T cells are activated after CVB3 infection of C57BL/6 and Bl.Tg.Ealpha mice. Depletion of gamma delta+ T cells abrogated myocarditis susceptibility in IE+ animals and resulted in a Th1 ->Th2 phenotype shift. These studies indicate that the MHC class II antigen haplotype controls myocarditis susceptibility, that this control is most likely mediated through the type of gamma delta T cells activated during CVB3 infection, and finally that different subpopulations of gamma delta+ T cells may either promote or inhibit Th1 cell responses. PMID- 10364313 TI - Identification of multiple protective epitopes (protectopes) in the central conserved domain of a prototype human respiratory syncytial virus G protein. AB - A recombinant fusion protein (BBG2Na) comprising the central conserved domain of the respiratory syncytial virus subgroup A (RSV-A) (Long) G protein (residues 130 to 230) and an albumin binding domain of streptococcal protein G was shown previously to protect mouse upper (URT) and lower (LRT) respiratory tracts against intranasal RSV challenge (U. F. Power, H. Plotnicky-Gilquin, T. Huss, A. Robert, M. Trudel, S. Stahl, M. Uhlen, T. N. Nguyen, and H. Binz, Virology 230:155-166, 1997). Panels of monoclonal antibodies (MAbs) and synthetic peptides were generated to facilitate dissection of the structural elements of this domain implicated in protective efficacy. All MAbs recognized native RSV-A antigens, and five linear B-cell epitopes were identified; these mapped to residues 152 to 163, 165 to 172, 171 to 187 (two overlapping epitopes), and 196 to 204, thereby covering the highly conserved cysteine noose domain. Antibody passive-transfer and peptide immunization studies revealed that all epitopes were implicated in protection of the LRT, but not likely the URT, against RSV-A challenge. Pepscan analyses of anti-RSV-A and anti-BBG2Na murine polyclonal sera revealed lower level epitope usage within the central conserved region in the former, suggesting diminished immunogenicity of the implicated epitopes in the context of the whole virus. However, Pepscan analyses of RSV-seropositive human sera revealed that all of the murine B-cell protective epitopes (protectopes) that mapped to the central conserved domain were recognized in man. Should these murine protectopes also be implicated in human LRT protection, their clustering around the highly conserved cysteine noose region will have important implications for the development of RSV vaccines. PMID- 10364314 TI - Nonhomologous RNA recombination in bovine viral diarrhea virus: molecular characterization of a variety of subgenomic RNAs isolated during an outbreak of fatal mucosal disease. AB - Four bovine viral diarrhea virus type 2 (BVDV-2) pairs consisting of cytopathogenic (cp) and noncp BVDV-2 were isolated during an outbreak of mucosal disease. Comparative sequence analysis showed that the four noncp BVDV-2 isolates were almost identical. For the cp BVDV-2 isolates, viral subgenomic RNAs were shown by Northern blot to have a length of about 8 kb, which is about 4.3 kb shorter than the genome of noncp BVDV. Cytopathogenicity and the expression of NS3 were both strictly correlated to the presence of viral subgenomic RNAs. By reverse transcription-PCR, Southern blot analysis, and nucleotide sequencing, a set of 11 unique subgenomes was identified with up to 5 different subgenomes isolated from one animal. To our knowledge, this is the first report on isolation of a set of pestiviral subgenomes from individual animals. Common features of the BVDV-2 subgenomic RNAs include (i) deletion of most of the genomic region encoding the structural proteins, as well as the nonstructural proteins p7 and NS2, and (ii) insertion of cellular (poly)ubiquitin coding sequences. Three subgenomes also comprised 15 to 75 nucleotides derived from the 5' part of the NS2 gene. Comparisons of the obtained nucleotide sequences revealed that the different BVDV-2 subgenomes evolved from the respective noncp BVDV-2 by RNA recombination. The presence of short regions of sequence similarity at several crossing-over sites suggests that base pairing between the nascent RNA strand and the acceptor RNA template facilitates template switching of the BVDV RNA dependent RNA polymerase. PMID- 10364316 TI - A recombinant human cytomegalovirus with a large deletion in UL97 has a severe replication deficiency. AB - Human cytomegalovirus encodes a protein kinase (UL97) that confers sensitivity to ganciclovir by phosphorylating it to the monophosphate. The function of this unusual kinase in viral replication is unknown. We constructed two independent isolates of a recombinant virus, RCDelta97, that contain large deletions in this gene and carry a 4.8-kb insertion containing a selectable genetic marker. These mutant viruses were isolated by using a population of primary cells (HEL97) that express this gene from integrated copies of a defective retroviral vector. The recombinant viruses were severely impaired in their ability to replicate in primary fibroblasts, attaining virus titers that were 2 to 3 orders of magnitude lower than those produced by the parent virus. Despite the severe replication deficit, both of these viruses retained the ability to form small, slowly growing plaques in primary fibroblasts, demonstrating that UL97 is not absolutely essential for replication in cell culture. The replication deficit was relieved when UL97 was provided in trans in the complementing cell line, showing that the phenotype was due to a deficiency in UL97. Thus, the UL97 gene product plays a very important role in viral replication in tissue culture and may be a good target for antiviral chemotherapy. PMID- 10364315 TI - Formation of virus assembly intermediate complexes in the cytoplasm by wild-type and assembly-defective mutant human immunodeficiency virus type 1 and their association with membranes. AB - We have previously identified two distinct forms of putative viral assembly intermediate complexes, a detergent-resistant complex (DRC) and a detergent sensitive complex (DSC), in human immunodeficiency virus type 1 (HIV-1)-infected CD4(+) T cells (Y. M. Lee and X. F. Yu, Virology 243:78-93, 1998). In the present study, the intracellular localization of these two viral assembly intermediate complexes was investigated by use of a newly developed method of subcellular fractionation. In wild-type HIV-1-infected H9 cells, the DRC fractionated with the soluble cytoplasmic fraction, whereas the DSC was associated with the membrane fraction. The DRC was also detected in the cytoplasmic fraction in H9 cells expressing HIV-1 Myr- mutant Gag. However, little of the unmyristylated Gag and Gag-Pol proteins was found in the membrane fraction. Furthermore, HIV-1 Gag proteins synthesized in vitro in a rabbit reticulocyte lysate system in the absence of exogenous lipid membrane were able to assemble into a viral Gag complex similar to that of the DRC identified in infected H9 cells. The density of the viral Gag complex was not altered by treatment with the nonionic detergent Triton X-100, suggesting a lack of association of this complex with endogenous lipid. Formation of the DRC was not significantly affected by mutations in assembly domains M and L of the Gag protein but was drastically inhibited by a mutation in the assembly I domain. Purified DRC could be disrupted by high-salt treatment, suggesting electrostatic interactions are important for stabilizing the DRC. The Gag precursor proteins in the DRC were more sensitive to trypsin digestion than those in the DSC. These findings suggest that HIV-1 Gag and Gag Pol precursors assemble into DRC in the cytoplasm, a process which requires the protein-protein interaction domain (I) in NCp7; subsequently, the DRC is transported to the plasma membrane through a process mediated by the M domain of the matrix protein. It appears that during this process, a conformational change might occur in the DRC either before or after its association with the plasma membrane, and this change is followed by the detection of virus budding structure at the plasma membrane. PMID- 10364317 TI - Appearance of mink cell focus-inducing recombinants during in vivo infection by moloney murine leukemia virus (M-MuLV) or the Mo+PyF101 M-MuLV enhancer variant: implications for sites of generation and roles in leukemogenesis. AB - One hallmark of murine leukemia virus (MuLV) leukemogenesis in mice is the appearance of env gene recombinants known as mink cell focus-inducing (MCF) viruses. The site(s) of MCF recombinant generation in the animal during Moloney MuLV (M-MuLV) infection is unknown, and the exact roles of MCF viruses in disease induction remain unclear. Previous comparative studies between M-MuLV and an enhancer variant, Mo+PyF101 MuLV, suggested that MCF generation or early propagation might take place in the bone marrow under conditions of efficient leukemogenesis. Moreover, M-MuLV induces disease efficiently following both intraperitoneal (i.p.) and subcutaneous (s.c.) inoculation but leukemogenicity by Mo+PyF101 M-MuLV is efficient following i.p. inoculation but attenuated upon s. c. inoculation. Time course studies of MCF recombinant appearance in the bone marrow, spleen, and thymus of wild-type and Mo+PyF101 M-MuLV i.p.- and s.c. inoculated mice were carried out by performing focal immunofluorescence assays. Both the route of inoculation and the presence of the PyF101 enhancer sequences affected the patterns of MCF generation or early propagation. The bone marrow was a likely site of MCF recombinant generation and/or early propagation following i.p. inoculation of M-MuLV. On the other hand, when the same virus was inoculated s.c., the primary site of MCF generation appeared to be the thymus. Also, when Mo+PyF101 M-MuLV was inoculated i.p., MCF generation appeared to occur primarily in the thymus. The time course studies indicated that MCF recombinants are not involved in preleukemic changes such as splenic hyperplasia. On the other hand, MCFs were detected in tumors from Mo+PyF101 M-MuLV s. c.-inoculated mice even though they were largely undetectable at preleukemic times. These results support a role for MCF recombinants late in disease induction. PMID- 10364318 TI - Inhibition of herpes simplex virus gD and lymphotoxin-alpha binding to HveA by peptide antagonists. AB - The herpesvirus entry mediator A (HveA) is a recently characterized member of the tumor necrosis factor receptor family that mediates the entry of most herpes simplex virus type 1 (HSV-1) strains into mammalian cells. Studies on the interaction of HSV-1 with HveA have shown that of all the viral proteins involved in uptake, only gD has been shown to bind directly to HveA, and this binding mediates viral entry into cells. In addition to gD binding to HveA, the latter has been shown to interact with proteins of tumor necrosis factor receptor associated factor family, lymphotoxin-alpha (LT-alpha), and a membrane-associated protein referred to as LIGHT. To study the relationship between HveA, its natural ligands, and the viral proteins involved in HSV entry into cells, we have screened two phage-displayed combinatorial peptide libraries for peptide ligands of a recombinant form of HveA. Affinity selection experiments yielded two peptide ligands, BP-1 and BP-2, which could block the interaction between gD and HveA. Of the two peptides, only BP-2 inhibited HSV entry into CHO cells transfected with an HveA-expressing plasmid. When we analyzed these peptides for the ability to interfere with HveA binding to its natural ligand LT-alpha, we found that BP-1 inhibited the interaction of cellular LT-alpha with HveA. Thus, we have dissected the sites of interaction between the cell receptor, its natural ligand LT-alpha and gD, the virus-specific protein involved in HSV entry into cells. PMID- 10364319 TI - Epstein-Barr virus nuclear antigen 3C interacts with histone deacetylase to repress transcription. AB - EBNA3C can specifically repress the expression of reporter plasmids containing EBV Cp latency-associated promoter elements. Cp is normally the main promoter for EBNA mRNA initiation, so it appears that EBNA3C contributes to a negative autoregulatory control loop. By mutational analysis it was previously established that this repression is consistent with EBNA3C being targeted to Cp by binding the cellular sequence-specific DNA-binding protein CBF1 (also known as recombination signal-binding protein [RBP]-Jkappa. Further analysis suggested that in vivo a corepressor interacts with EBNA3C in this DNA binding complex. Results presented here are all consistent with a component of such a corepressor exhibiting histone deacetylase activity. The drug trichostatin A, which specifically inhibits histone deacetylases, relieved two- to threefold the repression of Cp induced by EBNA3C in two different cell types. Moreover, repression of pTK-CAT-Cp4x by EBNA3C was specifically enhanced by cotransfection of an expression plasmid for human histone deacetylase-1 (HDAC1). Consistent with these functional assays, in vitro-translated HDAC1 bound to a glutathione S transferase (GST) fusion protein including full-length EBNA3C, and in the reciprocal experiment EBNA3C bound to a GST fusion with the N terminus of HDAC1. Coimmunoprecipitations also revealed an EBNA3C-HDAC1 interaction in vivo, and GST EBNA3C bound functional histone deacetylase enzyme activity from HeLa cell nuclear extracts. The region of EBNA3C involved in the interaction with HDAC1 appears to correspond to the region which is necessary for binding to CBF1/RBP Jkappa. A direct physical interaction between EBNA3C and HDAC1 was demonstrated with recombinant proteins purified from bacterial cells, and we therefore conclude that HDAC1 and CBF1/RBP-Jkappa bind to the same or adjacent regions of EBNA3C. These data suggest that recruitment of histone deacetylase activity makes a significant contribution to the repression of transcription from Cp because EBNA3C bridges an interaction between CBF1/RBP-Jkappa and HDAC1. PMID- 10364320 TI - Variable constraints on the principal immunodominant domain of the transmembrane glycoprotein of human immunodeficiency virus type 1. AB - Lentiviruses have in their transmembrane glycoprotein (TM) a highly immunogenic structure referred to as the principal immunodominant domain (PID). The PID forms a loop of 5 to 7 amino acids between two conserved cysteines. Previous studies showed that envelope (Env) glycoprotein functions of feline immunodeficiency virus (FIV) could be retained after extensive mutation of the PID loop sequence, in spite of its high conservation. In order to compare Env function in different lentiviruses, either random mutations were introduced in the PID loop sequence of human immunodeficiency virus type 1 (HIV-1) or the entire HIV-1 PID loop was replaced by the corresponding PID loop of FIV or simian immunodeficiency virus (SIV). In the macrophage-tropic HIV-1 ADA Env, mutations impaired the processing of the gp160 Env precursor, thereby abolishing viral infectivity. However, 6 of the 108 random Env mutants that were screened retained the capacity to induce cell membrane fusion. The SIV and FIV sequences and five random mutations were then introduced in the context of T-cell-line-adapted HIV-1 LAI which, although phenotypically distant from HIV-1 ADA, has an identical PID loop sequence. In contrast to the situation for HIV-1 ADA mutants, the cleavage of the Env precursor was unaffected in most HIV-1 LAI mutants. Such mutations, however, resulted in increased shedding of the gp120 surface glycoprotein (SU) from the gp41 TM. The HIV-1 LAI Env mutants showed high fusogenic efficiency. Three Env mutants retained the capacity to mediate virus entry in target cells, although less efficiently than the wild-type Env, and allowed the reconstitution of infectious molecular clones. These results indicated that in HIV-1, like FIV, the conserved PID sequence can be changed without impairing Env function. However, functional constraints on the PID of HIV-1 vary depending on the structural context of Env, presumably in relation to the role of the PID in the interaction of the SU and TM subunits and the stability of the Env complex. PMID- 10364321 TI - Both neutralization resistance and high infectivity phenotypes are caused by mutations of interacting residues in the human immunodeficiency virus type 1 gp41 leucine zipper and the gp120 receptor- and coreceptor-binding domains. AB - Neutralization resistance of human immunodeficiency virus type 1 (HIV-1) is a major impediment to vaccine development. We have found that residues of HIV-1 MN strain in the C terminus of gp120 and the leucine zipper (LZ) region of gp41 viral envelope proteins interact cooperatively to determine neutralization resistance and modulate infectivity. Further, results demonstrate that this interaction, by which regions of gp120 are assembled onto the LZ, involves amino acid residues intimately related to those which participate in the binding of the envelope to its receptor and coreceptor. Variations in this critical assembly structure determine the concordant, interdependent evolution of increased infectivity efficiency and neutralization resistance phenotypes of the envelopes. The results elucidate important structure-function relationships among epitopes that are important targets of vaccine development. PMID- 10364322 TI - Packaging-competent capsids of a herpes simplex virus temperature-sensitive mutant have properties similar to those of in vitro-assembled procapsids. AB - Newcomb and coworkers (W. W. Newcomb, F. L. Homa, D. R. Thomsen, F. P. Booy, B. L. Trus, A. C. Steven, J. V. Spencer, and J. C. Brown, J. Mol. Biol. 263:432-446, 1996; W. W. Newcomb, F. L. Homa, D. R. Thomsen, Z. Ye, and J. C. Brown, J. Virol. 68:6059-6063, 1994) have recently described an in vitro herpes simplex virus (HSV) capsid assembly product which, because of certain parallels between its properties and those of bacteriophage proheads, they have designated the procapsid. As in their bacteriophage counterparts, there are marked differences between the structures of the two types of particle, and conversion from the procapsid to the capsid form requires extensive reconfiguration of the subunits. This reconfiguration occurs spontaneously upon extended in vitro incubation. One of the distinctive features of the HSV procapsids is that, unlike mature capsids, they are unstable and disassemble upon storage at 2 degrees C. Using a mutant of HSV type 1 (ts1201), which has a lesion in the protease responsible for maturational cleavage of the scaffolding protein, we have demonstrated that capsids present within cells infected at nonpermissive temperatures are also cryosensitive and disappear if the cells are incubated at 0 degrees C. This suggests that ts1201 capsids may resemble procapsids in structure. However, ts1201 capsids remain cryosensitive following extended incubation at an elevated temperature and, therefore, do not appear to undergo the spontaneous reconfiguration seen with in vitro-assembled procapsids. The lesion in ts1201 is reversible, and capsids formed at the nonpermissive temperature can undergo maturational cleavage and go on to form infectious virions following downshift to permissive temperatures. The sensitivity of ts1201 capsids to low temperatures is closely correlated with the cleavage status of the scaffolding protein, suggesting that proteolysis may act to trigger their conversion to the stable form. The experiments described here provide the firmest evidence yet that the procapsid has a biologically relevant role in the virus life cycle. PMID- 10364323 TI - A complex translational program generates multiple novel proteins from the latently expressed kaposin (K12) locus of Kaposi's sarcoma-associated herpesvirus. AB - The most abundantly expressed latent transcripts encoded by the Kaposi's sarcoma (KS)-associated herpesvirus derive from the genomic region surrounding open reading frame (ORF) K12 (kaposin A). Here we show that these transcripts, initially described as limited to ORF K12 itself, more frequently encompass upstream sequences spanning two sets of 23-nucleotide GC-rich direct repeats (DRs) (DR1 and DR2). Although the DRs lack AUG codons and were previously presumed to be noncoding, a monoclonal antibody raised to infected cells detected multiple polypeptides encoded by this region. These proteins are expressed during latency and upon induction of lytic viral replication in both primary effusion lymphoma (PEL) cell lines and KS tumors. Biochemical and genetic analyses reveal that these proteins are derived from variant translational initiation at CUG codons. The predominant translation product in the PEL cell line BCBL-1 derives from the 5'-most CUG codon in the transcript, resulting in a protein (termed kaposin B) which is encoded largely by the repeats themselves and which does not include K12 sequences. Other non-AUG codons in alternate reading frames are also used at lower efficiency, including one that initiates translation of a DR-K12 fusion protein (kaposin C) that is predicted to sort to a different subcellular locale than kaposin B. Thus, the products of the K12 region, which is the most abundantly transcribed region in latency, are surprisingly complex and may encompass multiple biological functions. PMID- 10364324 TI - The mechanism of an immature secretion phenotype of a highly frequent naturally occurring missense mutation at codon 97 of human hepatitis B virus core antigen. AB - A very frequent missense mutation at codon 97 of human hepatitis B virus (HBV) core antigen (HBcAg) has been found in chronic carriers worldwide. Functional characterization of this mutant revealed one intracellular and two extracellular phenotypes in contrast to wild-type HBV: (i) a 6- to 12-fold decrease in the level of the full-length relaxed circular DNA, a 4- to 5-fold decrease in the plus-strand DNA, and an approximately 1.8-fold decrease in the minus-strand and overall DNA levels in the intracellular viral core particles; (ii) a 5.7-fold increase in the immature secretion of Dane particles, containing minus-strand, single-stranded virion DNA; and (iii) a significant reduction of nonenveloped core particles in the medium. The steady-state levels of mutant and wild-type core proteins expressed from the same vector appeared to be similar. Using a complementation assay and gradient centrifugation analysis, we demonstrated that this mutant core protein alone is necessary and sufficient for immature secretion. The decreased level of intracellular HBV DNA is caused by both the cis defect of the mutant genome and the trans defect of the mutant core protein. We have dissected further the relationship between the intracellular and extracellular phenotypes of mutant F97L. The pleiotropic effects of the HBcAg codon 97 mutation were observed consistently in several different experimental settings. The mechanism and biological significance of these findings are discussed. PMID- 10364325 TI - Selection and characterization of human immunodeficiency virus type 1 mutants that are resistant to inhibition by the transdominant negative RevM10 protein. AB - Intracellular immunization with RevM10, a transdominant negative form of the Rev protein, efficiently inhibits human immunodeficiency virus (HIV) replication in vitro and gene therapy protocols that use this modality are currently being evaluated in human clinical trials. Development of resistance to this kind of therapy has not been previously reported. Here we show that RevM10-resistant HIV type 1 (HIV-1) variants can be selected by in vitro passage of HIV-1 in a T lymphoblastoid cell line constitutively expressing RevM10. Unexpectedly, the selected variants showed changes in the Rev response element (RRE) but no changes in Rev. Replacement of the wild-type RRE with a mutated RRE resulted in a virus that showed increased resistance to RevM10. After repeated passages of the resistant variant in cells expressing RevM10, a virus with an additional mutation in the viral vpu gene was selected. Surprisingly, a virus containing only this vpu mutation also showed some resistance to inhibition by RevM10. PMID- 10364326 TI - Differential regulation of Dk and Kk major histocompatibility complex class I proteins on the cell surface after infection of murine cells by pseudorabies virus. AB - After pseudorabies virus (PRV) infection of murine L929 cells, the cell surface expression of major histocompatibility complex (MHC) class I proteins changes such that the total amount of MHC class I molecules remains relatively constant but the levels of the individual alleles Dk and Kk vary. This is an active process involving at least three PRV gene products that act in an allele-specific manner such that cell surface expression of MHC class I Dk is decreased and that of Kk is increased. Our results indicate that an early gene product mediates the overall reduction in Dk protein and a late gene product which is mutant in the attenuated PRV strain Bartha mediates the increase in Kk protein. We provide additional evidence for a third gene product involved in the regulation of the synthesis of both the Dk and Kk proteins. In addition, we show that the early decrease in the Dk protein is not due to a block in synthesis or processing of the complex through the secretory system. PMID- 10364327 TI - DNA microarrays of the complex human cytomegalovirus genome: profiling kinetic class with drug sensitivity of viral gene expression. AB - We describe, for the first time, the generation of a viral DNA chip for simultaneous expression measurements of nearly all known open reading frames (ORFs) in the largest member of the herpesvirus family, human cytomegalovirus (HCMV). In this study, an HCMV chip was fabricated and used to characterize the temporal class of viral gene expression. The viral chip is composed of microarrays of viral DNA prepared by robotic deposition of oligonucleotides on glass for ORFs in the HCMV genome. Viral gene expression was monitored by hybridization to the oligonucleotide microarrays with fluorescently labelled cDNAs prepared from mock-infected or infected human foreskin fibroblast cells. By using cycloheximide and ganciclovir to block de novo viral protein synthesis and viral DNA replication, respectively, the kinetic classes of array elements were classified. The expression profiles of known ORFs and many previously uncharacterized ORFs provided a temporal map of immediate-early (alpha), early (beta), early-late (gamma1), and late (gamma2) genes in the entire genome of HCMV. Sequence compositional analysis of the 5' noncoding DNA sequences of the temporal classes, performed by using algorithms that automatically search for defined and recurring motifs in unaligned sequences, indicated the presence of potential regulatory motifs for beta, gamma1, and gamma2 genes. In summary, these fabricated microarrays of viral DNA allow rapid and parallel analysis of gene expression at the whole viral genome level. The viral chip approach coupled with global biochemical and genetic strategies should greatly speed the functional analysis of established as well as newly discovered large viral genomes. PMID- 10364328 TI - La autoantigen specifically recognizes a predicted stem-loop in hepatitis B virus RNA. AB - We recently identified three nuclear proteins (p45, p39, and p26) that bind to a 91-nucleotide (nt) RNA element between nt 1243 and 1333 in hepatitis B virus (HBV) RNA, and we showed that these proteins and HBV RNA are regulated coordinately by gamma interferon and tumor necrosis factor alpha. Purification and sequence analysis of tryptic peptides obtained from p39 revealed sequence homology to the mouse La protein. Immunoprecipitation experiments showed that p45, p39, and p26 were recognized by anti-La-specific antiserum, indicating that p45 is the full-length La protein and that p39 and p26 are likely to be proteolytic La cleavage products. Furthermore, in competition experiments we found that all three La proteins bind, in a phosphorylation-dependent manner, to the same predicted stem-loop structure located between nt 1275 and 1291 of HBV, with Kds of approximately 1.0 nM. Collectively, these results support the notion that the La protein may contribute to HBV RNA stability, constitutively and in response to inflammatory cytokines. PMID- 10364329 TI - Analysis of the effect of natural sequence variation in Tat and in cyclin T on the formation and RNA binding properties of Tat-cyclin T complexes. AB - The biological activity of the human immunodeficiency virus type 1 (HIV-1) Tat (Tat1) transcriptional activator requires the recruitment of a Tat1-CyclinT1 (CycT1) complex to the TAR RNA target encoded within the viral long terminal repeat (LTR). While other primate immunodeficiency viruses, such as HIV-2 and mandrill simian immunodeficiency virus (SIVmnd), also encode Tat proteins that activate transcription via RNA targets, these proteins differ significantly, both from each other and from Tat1, in terms of their ability to activate transcription directed by LTR promoter elements found in different HIV and SIV isolates. Here, we show that CycT1 also serves as an essential cofactor for HIV-2 Tat (Tat2) and SIVmnd Tat (Tat-M) function. Moreover, the CycT1 complex formed by each Tat protein displays a distinct RNA target specificity that accurately predicts the level of activation observed with a particular LTR. While Tat2 and Tat-M share the ability of Tat1 to bind to CycT1, they differ from Tat1 in that they are also able to bind to the related but distinct CycT2. However, the resultant Tat-CycT2 complexes fail to bind TAR and are therefore abortive. Surprisingly, mutation of a single residue in CycT2 (asparagine 260 to cysteine) rescues the ability of CycT2 to bind Tat1 and also activates not only TAR binding by all three Tat-CycT2 complexes but also Tat function. Therefore, the RNA target specificity of different Tat-CycT1 complexes is modulated by natural sequence variation in both the viral Tat transcriptional activator and in the host cell CycT molecule recruited by Tat. Further, the RNA target specificity of the resultant Tat-CycT1 complex accurately predicts the ability of that complex to activate transcription from a given LTR promoter element. PMID- 10364330 TI - Structural constraints on RNA virus evolution. AB - The recently discovered hepatitis G virus (HGV) or GB virus C (GBV-C) is widely distributed in human populations, and homologues such as HGV/GBV-CCPZ and GBV-A are found in a variety of different primate species. Both epidemiological and phylogenetic analyses support the hypothesis that GB viruses coevolved with their primate hosts, although their degree of sequence similarity appears incompatible with the high rate of sequence change of HGV/GBV-C over short observation periods. Comparison of complete coding sequences (8,500 bases) of different genotypes of HGV/GBV-C showed an excess of invariant synonymous sites (at 23% of all codons) compared with the frequency expected by chance (10%). To investigate the hypothesis that RNA secondary-structure formation through internal base pairing limited sequence variability at these sites, an algorithm was developed to detect covariant sites among HGV/GBV-C sequences of different genotypes. At least 35 covariant sites that were spatially associated with potential stem-loop structures were detected, whose positions correlated with positions in the genome that showed reductions in synonymous variability. Although the functional roles of the predicted secondary structures remain unclear, the restriction of sequence change imposed by secondary-structure formation provides a mechanism for differences in net rate of accumulation of nucleotide substitutions at different sites. However, the resulting disparity between short- and long-term rates of sequence change of HGV/GBV-C violates the assumptions of the "molecular clock." This places a major restriction on the use of nucleotide or amino acid sequence comparisons to calculate times of divergence of other viruses evolving under the same structural constraints as GB viruses. PMID- 10364331 TI - Peptide ligands to human immunodeficiency virus type 1 gp120 identified from phage display libraries. AB - We have used phage-displayed peptide libraries to identify novel ligands to the human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein gp120. Screening of libraries of random 12-mers, 7-mers, and cyclic 9-mers produced two families of gp120 binding peptides. Members of a family with the prototype sequence RINNIPWSEAMM (peptide 12p1) inhibit the interaction between gp120 and both four-domain soluble CD4 (4dCD4) and monoclonal antibody (MAb) 17b, a neutralizing antibody that covers the chemokine receptor binding surface on gp120. Peptide 12p1 inhibits the interaction of 4dCD4 with gp120 from three different HIV strains, implying that it binds to a conserved site on gp120. Members of a second family of peptides, with the prototype sequence TSPYEDWQTYLM (peptide 12p2), bind more weakly to gp120. They do not detectably affect its interaction with 4dCD4, but they enhance its binding to MAb 17b. A common sequence motif in the two peptide families and cross-competition for gp120 binding suggest that they have overlapping contacts. Their divergent effects on the affinity of gp120 for MAb 17b may indicate that their binding stabilizes distinct conformational states of gp120. The functional properties of 12p1 suggest that it might be a useful lead for the development of inhibitors of HIV entry. PMID- 10364332 TI - The P236L delavirdine-resistant human immunodeficiency virus type 1 mutant is replication defective and demonstrates alterations in both RNA 5'-end- and DNA 3' end-directed RNase H activities. AB - The nonnucleoside reverse transcriptase (RT) inhibitor (NNRTI) delavirdine (DLV) selects in vitro for the human immunodeficiency virus type 1 (HIV-1) RT mutation P236L, which confers high-level resistance to DLV but not other NNRTIs. Unexpectedly, P236L has developed infrequently in HIV-1 isolates obtained from patients receiving DLV; K103N is the predominant resistance mutation observed in that setting. We characterized the replication fitness of viruses derived from pNL4-3 containing P236L or K103N in both H9 and primary human peripheral blood mononuclear cell cultures infected in parallel with the two mutants. In the absence of DLV, p24 production by wild-type virus occurred more rapidly and to higher levels than with either mutant; P236L consistently demonstrated a two- to threefold decrease in p24 relative to K103N. At low levels of DLV, growth of wild type virus was severely inhibited, and K103N replicated two- to threefold more efficiently than P236L. At high concentrations of DLV, P236L replication and K103N replication were both inhibited. Recombinant RTs containing K103N or P236L were analyzed for DNA polymerization on heteropolymeric RNA templates and RNase H degradation of RNA-DNA hybrids. Neither mutant demonstrated defects in polymerization. K103N demonstrated normal RNA 5'-end-directed RNase H cleavage and slowed DNA 3'-end-directed RNase H cleavage compared to wild-type RT. P236L demonstrated slowing of both DNA 3'-end- and RNA 5'-end-directed RNase H cleavage, consistent with its reduced replication efficiency relative to K103N. These data suggest that NNRTI resistance mutations can lead to reductions in the efficiency of RNase H cleavage, which may contribute to a reduction in the replication fitness of HIV-1. PMID- 10364333 TI - Induction of AIDS in rhesus monkeys by a recombinant simian immunodeficiency virus expressing nef of human immunodeficiency virus type 1. AB - A nef gene is present in all primate lentiviruses, including human immunodeficiency virus type 1 (HIV-1) and simian immunodeficiency virus of macaque monkeys (SIVmac). However, the nef genes of HIV-1 and SIVmac exhibit minimal sequence identity, and not all properties are shared by the two. Nef sequences of SIVmac239 were replaced by four independent nef alleles of HIV-1 in a context that was optimal for expression. The sources of the HIV-1 nef sequences included NL 4-3, a variant NL 4-3 gene derived from a recombinant-infected rhesus monkey, a patient nef allele, and a nef consensus sequence. Of 16 rhesus monkeys infected with these SHIVnef chimeras, 9 maintained high viral loads for prolonged periods, as observed with the parental SIVmac239, and 6 have died with AIDS 52 to 110 weeks postinfection. Persistent high loads were observed at similar frequencies with the four different SIV recombinants that expressed these independent HIV-1 nef alleles. Infection with other recombinant SHIVnef constructions resulted in sequence changes in infected monkeys that either created an open nef reading frame or optimized the HIV-1 nef translational context. The HIV-1 nef gene was uniformly retained in all SHIVnef-infected monkeys. These results demonstrate that HIV-1 nef can substitute for SIVmac nef in vivo to produce a pathogenic infection. However, the model suffers from an inability to consistently obtain persisting high viral loads in 100% of the infected animals, as is observed with the parental SIVmac239. PMID- 10364334 TI - Discrimination between sialic acid-containing receptors and pseudoreceptors regulates polyomavirus spread in the mouse. AB - Variations in the polyomavirus major capsid protein VP1 underlie important biological differences between highly pathogenic large-plaque and relatively nonpathogenic small-plaque strains. These polymorphisms constitute major determinants of virus spread in mice and also dictate previously recognized strain differences in sialyloligosaccharide binding. X-ray crystallographic studies have shown that these determinants affect binding to the sialic acids. Here we report results of further experiments designed to test the importance of specific contacts between VP1 and the carbohydrate moieties of the receptor. With minor exceptions, substitutions at positions predicted from crystallography to be important in binding the terminal alpha-2,3-linked sialic acid or the penultimate sugar (galactose) destroyed the ability of the virus to replicate in cell culture. Substitutions that prevented binding to a branched disialyloligosaccharide were found to result in viruses that were both viable in culture and tumorigenic in the mouse. Conversely, substitutions that allowed recognition and binding of the branched carbohydrate chain inhibited spread in the mouse, though the viruses remained viable in culture. Mice of five different inbred strains, all highly susceptible to large-plaque virus, showed resistance to the spread of polyomavirus strains bearing the VP1 type which binds the branched-chain receptor. We suggest that glycoproteins bearing the appropriate O linked branched sialyloligosaccharide chains are effective pseudoreceptors in the host and that they block the spread of potentially tumorigenic or virulent virus strains. PMID- 10364335 TI - Dendritic cell-T-cell interactions support coreceptor-independent human immunodeficiency virus type 1 transmission in the human genital tract. AB - Worldwide, human immunodeficiency virus (HIV) is transmitted predominantly by heterosexual contact. Here, we investigate for the first time, by examining mononuclear cells obtained from cervicovaginal tissue, the mechanisms whereby HIV type 1 (HIV-1) directly targets cells from the human genital tract. In contrast to earlier findings in mucosal models such as human skin, we demonstrate that the majority of T cells and macrophages but none or few dendritic cells (DC) express the HIV-1 coreceptor CCR5 in normal human cervicovaginal mucosa, whereas all three cell types express the coreceptor CXCR4. To understand the role of coreceptor expression on infectivity, mucosal mononuclear cells were infected with various HIV-1 isolates, using either CCR5 or CXCR4. Unstimulated T cells become rapidly, albeit nonproductively, infected with R5- and X4-tropic variants. However, DC and T cells form stable conjugates which permit productive infection by viruses of both coreceptor specificities. These results indicate that HIV-1 can exploit T-cell-DC synergism in the human genital tract to overcome potential coreceptor restrictions on DC and postentry blocks of viral replication in unactivated T cells. Thus, mononuclear cells infiltrating the genital mucosa are permissive for transmission of both R5- and X4-tropic HIV-1 variants, and selection of virus variants does not occur by differential expression of HIV-1 coreceptors on genital mononuclear cells. PMID- 10364336 TI - Evidence of avian leukosis virus subgroup E and endogenous avian virus in measles and mumps vaccines derived from chicken cells: investigation of transmission to vaccine recipients. AB - Reverse transcriptase (RT) activity has been detected recently in all chicken cell-derived measles and mumps vaccines. A study of a vaccine manufactured in Europe indicated that the RT is associated with particles containing endogenous avian retrovirus (EAV-0) RNA and originates from the chicken embryonic fibroblasts (CEF) used as a substrate for propagation of the vaccine. We investigated the origin of RT in measles and mumps vaccines from a U.S. manufacturer and confirm the presence of RT and EAV RNA. Additionally, we provide new evidence for the presence of avian leukosis virus (ALV) in both CEF supernatants and vaccines. ALV pol sequences were first identified in particle associated RNA by amplification with degenerate retroviral pol primers. ALV RNA sequences from both the gag and env regions were also detected. Analysis of hypervariable region 2 of env revealed a subgroup E sequence, an endogenous-type ALV. Both CEF- and vaccine-derived RT activity could be blocked by antibodies to ALV RT. Release of ALV-like virus particles from uninoculated CEF was also documented by electron microscopy. Nonetheless, infectivity studies on susceptible 15B1 chicken cells gave no evidence of infectious ALV, which is consistent with the phenotypes of the ev loci identified in the CEF. PCR analysis of ALV and EAV proviral sequences in peripheral blood mononuclear cells from 33 children after measles and mumps vaccination yielded negative results. Our data indicate that the sources of RT activity in all RT-positive measles and mumps vaccines may not be similar and depend on the particular endogenous retroviral loci present in the chicken cell substrate used. The present data do not support transmission of either ALV or EAV to recipients of the U.S.-made vaccine and provide reassurance for current immunization policies. PMID- 10364337 TI - Role of the M2-1 transcription antitermination protein of respiratory syncytial virus in sequential transcription. AB - M2-1 protein of human respiratory syncytial virus (RSV) is a transcription antitermination factor that is important for the efficient synthesis of full length mRNAs as well as for the synthesis of polycistronic readthrough mRNAs, which are characteristic of nonsegmented negative-strand RNA viruses. The contributions of these effects to RSV sequential transcription were investigated with minigenomes which contained one to five genes which were either foreign marker genes or authentic RSV genes. When evaluated on a promoter-proximal gene, the effect of M2-1 on the synthesis of full-length mRNA was much greater for a long (1,212- or 1,780-nucleotide) gene (up to a 615-fold increase) than for a short (274-nucleotide) gene (less than a 2-fold increase). This was independent of whether the gene contained non-RSV or RSV-specific sequence. Once the polymerase had terminated prematurely, it was unable to reinitiate at a downstream gene. These studies also confirmed that M2-1 enhances the synthesis of polycistronic mRNAs and that the magnitude of this effect varied greatly among different naturally occurring gene junctions. The synthesis of polycistronic mRNAs, which presumably involves antitermination at the gene-end signal, required a higher level of M2-1 than did the synthesis of the corresponding monocistronic mRNAs. M2-1 did not have a comparable antitermination effect at the junction between the leader region and the first gene. In a minigenome containing the NS1 and NS2 genes in their authentic sequence context, synthesis of full-length NS1 and NS2 mRNAs in the absence of M2-1 was remarkably high (36 and 57%, respectively, of the maximum levels observed in the presence of M2-1). In contrast, synthesis of mRNA from additional downstream genes was highly dependent on M2-1. Thus, RSV has the potential for two transcription programs: one in the absence of M2-1, in which only the NS1 and NS2 genes are transcribed, and one in the presence of M2-1, in which sequential transcription of the complete genome occurs. The dependence on M2-1 for transcription was greater for a gene in the fifth position from the promoter than for one in the third position. This indicates that under conditions where M2-1 is limiting, its concentration affects the gradient of transcription. Although M2-1 was found to have profound effects on transcription, it had no effect on replication of any minigenome tested, suggesting that it is not an active participant in RNA replication or regulation of RNA replication. Finally, since a permissive RSV infection is marked by a gradual increase in the intracellular accumulation of viral proteins including M2 1, we examined the relative abundances of various mRNAs during RSV infection for evidence of temporal regulation of transcription. None was found, implying that the availability of M2-1 during a permissive infection is sufficient at all times such that its concentration does not mediate temporal regulation of gene transcription. PMID- 10364338 TI - Expression and use of human immunodeficiency virus type 1 coreceptors by human alveolar macrophages. AB - Human immunodeficiency virus type 1 (HIV-1) requires, in addition to CD4, coreceptors of the CC or CXC chemokine families for productive infection of T cells and cells of the monocyte-macrophage lineage. Based on the hypothesis that coreceptor expression on alveolar macrophages (AM) may influence HIV-1 infection of AM in the lung, this study analyzes the expression and utilization of HIV-1 coreceptors on AM of healthy individuals. AM were productively infected with five different primary isolates of HIV-1. Levels of surface expression of CCR5, CXCR4, and CD4 were low compared to those of blood monocytes, but CCR3 was not detectable. mRNA for CCR5, CXCR4, CCR2, and CCR3 were all detectable, but to varying degrees and with variability among donors. Expression of CCR5, CXCR4, and CCR2 mRNA was downregulated following stimulation with lipopolysaccharide (LPS). In contrast, secretion of the chemokines RANTES, MIP-1alpha, and MIP-1beta was upregulated with LPS stimulation. Interestingly, HIV-1 replication was diminished following LPS stimulation. Infection of AM with HIV-1 in the presence of the CC chemokines demonstrated blocking of infection. Together, these studies demonstrate that AM can be infected by a variety of primary HIV-1 isolates, AM express a variety of chemokine receptors, the dominant coreceptor used for HIV entry into AM is CCR5, the expression of these receptors is dependent on the state of activation of AM, and the ability of HIV-1 to infect AM may be modulated by expression of the chemokine receptors and by chemokines per se. PMID- 10364339 TI - Functional analysis of carboxy-terminal deletion mutants of c-Myb. AB - The c-myb gene is implicated in the differentiation and proliferation of hematopoietic cells. Truncations of the N and/or C terminus of c-Myb, found in v Myb, can potentiate its transforming ability. Two negative regulatory subregions, located in the C terminus, were mapped previously by using GAL4-c-Myb fusion proteins in transient transfection assays for the transcriptional activation of a GAL4-responsive reporter gene. To dissect the C terminus of c-Myb in terms of its involvement in transcriptional activation and oncogenic transformation, a series of C-terminal deletion mutants of c-Myb were analyzed. In addition, linker insertion mutants within the transactivation domain and/or heptad leucine repeat of c-Myb were examined along with those deletion mutants. In this study, we demonstrated that the removal of both of the two previously mapped negative regulatory subregions from the native form of c-Myb not only supertransactivates a Myb-responsive reporter gene but also potentiates its transforming ability in culture. However, in contrast to previous results, cells transformed by all of the mutants analyzed here except v-Myb itself exhibited the same phenotype as those transformed by c-Myb. The proliferating cells were bipotenial and differentiated into both the granulocytic and monocytic lineages. This result implies that the C terminus of c-Myb alone has no effect on the lineage determination. Finally, the transactivation activities of these mutants correlated with their transforming activities when a mim-1 reporter gene was used but not when a model promoter containing five tandem Myb-binding sites was used. In particular, a very weakly transforming mutant with a linker insertion in the heptad leucine repeat superactivated the model promoter but not the mim-1 reporter gene. PMID- 10364340 TI - The human papillomavirus type 16 E6 gene alone is sufficient to induce carcinomas in transgenic animals. AB - High-risk human papillomaviruses (HPVs) are the causative agents of certain human cancers. HPV type 16 (HPV16) is the papillomavirus most frequently associated with cervical cancer in women. The E6 and E7 genes of HPV are expressed in cells derived from these cancers and can transform cells in tissue culture. Animal experiments have demonstrated that E6 and E7 together cause tumors. We showed previously that E6 and E7 together or E7 alone could induce skin tumors in mice when these genes were expressed in the basal epithelia of the skin. In this study, we investigated the role that the E6 gene plays in carcinogenesis. We generated K14E6 transgenic mice, in which the HPV16 E6 gene was directed in its expression by the human keratin 14 promoter (hK14) to the basal layer of the epidermis. We found that E6 induced cellular hyperproliferation and epidermal hyperplasia and caused skin tumors in adult mice. Interestingly, the tumors derived from E6 were mostly malignant, as opposed to the tumors from E7 mice, which were mostly benign. This result leads us to hypothesize that E6 may contribute differently than E7 to HPV-associated carcinogenesis; whereas E7 primarily contributes to the early stages of carcinogenesis that lead to the formation of benign tumors, E6 primarily contributes to the late stages of carcinogenesis that lead to malignancy. PMID- 10364341 TI - Identification and rapid quantification of early- and late-lytic human herpesvirus 8 infection in single cells by flow cytometric analysis: characterization of antiherpesvirus agents. AB - Human herpesvirus 8 (HHV-8) infection is associated with Kaposi's sarcoma, primary effusion lymphoma (PEL), and multicentric Castleman's disease. In this study, we used monoclonal antibodies (MAbs) directed against HHV-8 lytic cycle associated proteins encoded by open reading frame (ORF) 59 (nuclear PF-8 protein) and ORF K8.1 (viral envelope glycoprotein K8.1 [gpK8.1]) to investigate HHV-8 lytic infection in single cells. Lytically infected cells were labeled with MAbs, stained with fluorescently conjugated secondary Abs, and analyzed by flow cytometry. A 3-day stimulation of HHV-8-positive PEL cell lines (BCBL-1 and BC-3) with 12-O-tetradecanoylphorbol-13-acetate (30 nM) or n-butyric acid (0.3 mM) maximized the expression of lytic-phase viral proteins and minimized cell toxicity. The absolute number of expressing cells was inducer and cell line dependent. Expression of PF-8 occurred earlier and more frequently (in up to 20% of cells) than did expression of gpK8.1. A subset of PF-8 positive cells (25%) co expressed gpK8.1, representing the majority of gpK8.1 expressing cells. Acyclovir, foscarnet, cidofovir, and PMEA reduced the number of cells expressing gpK8.1, but not the number expressing the nonstructural early lytic gene product PF-8. By contrast, alpha interferon (IFN-alpha) and IFN-beta reduced expression of both PF-8 and gpK8.1, implying an overall inhibitory effect on viral gene transcription or translation. In summary, we have characterized and quantified HHV-8 lytic infection in single cells by dual measurement of early- and late lytic-cycle HHV-8 protein expression. This technique should prove useful for screening of possible antiherpesvirus agents and for detailed phenotypic characterization of HHV-8-infected cells in vitro and in patients with HHV-8 associated diseases. PMID- 10364342 TI - A mouse model for the evaluation of pathogenesis and immunity to influenza A (H5N1) viruses isolated from humans. AB - During 1997 in Hong Kong, 18 human cases of respiratory illness, including 6 fatalities, were caused by highly pathogenic avian influenza A (H5N1) viruses. Since H5 viruses had previously been isolated only from avian species, the outbreak raised questions about the ability of these viruses to cause severe disease and death in humans. To better understand the pathogenesis and immunity to these viruses, we have used the BALB/c mouse model. Four H5N1 viruses replicated equally well in the lungs of mice without prior adaptation but differed in lethality for mice. H5N1 viruses that were highly lethal for mice were detected in multiple organs, including the brain. This is the first demonstration of an influenza A virus that replicates systemically in a mammalian species and is neurotropic without prior adaptation. The mouse model was also used to evaluate a strategy of vaccination against the highly pathogenic avian H5N1 viruses, using an inactivated vaccine prepared from nonpathogenic A/Duck/Singapore-Q/F119-3/97 (H5N3) virus that was antigenically related to the human H5N1 viruses. Mice administered vaccine intramuscularly, with or without alum, were completely protected from lethal challenge with H5N1 virus. Protection from infection was also observed in 70% of animals administered vaccine alone and 100% of mice administered vaccine with alum. The protective effect of vaccination correlated with the level of virus-specific serum antibody. These results suggests a strategy of vaccine preparedness for rapid intervention in future influenza pandemics that uses antigenically related nonpathogenic viruses as vaccine candidates. PMID- 10364343 TI - Recombination between two identical sequences within the same retroviral RNA molecule. AB - As a consequence of being diploid viruses, members of the Retroviridae have a high recombination rate. To measure recombination between two identical sequences within the same RNA molecule per round of retroviral replication cycle, a murine leukemia virus based vector (JZ442 + 3' Hyg) has been constructed. It carries a drug resistance gene, hyg, and a 290-bp repeat sequence of the 3' hyg gene inserted into the 3' untranslated region of the green fluorescent protein gene (gfp). Under fluorescence microscopy, Hygr cells containing the recombinant proviruses were clear, while a green color was observed in the drug-resistant cells carrying the parental proviruses. The rate of recombination was determined by the ratio of the number of clear colonies to the total number of Hygr colonies (green and clear colonies). The rate of recombination was found to be 62% by this method. The intermolecular recombination rate between an infectious virus bearing two copies of the 290-bp segment and a noninfectious chimeric RNA virus containing only a single copy of this sequence was also measured. PMID- 10364344 TI - Thymic tolerance to only one viral protein reduces lymphocytic choriomeningitis virus-induced immunopathology and increases survival in perforin-deficient mice. AB - The outcome of viral infections is dependent on the amount of tissue destruction caused either by direct lysis of infected cells and/or by immunopathology resulting from the immune response to the virus. We investigated whether induction of tolerance to only one viral protein could reduce immunopathology caused by nonlytic lymphocytic choriomeningitis virus (LCMV) in perforin deficient hosts. Earlier studies had shown that LCMV infection results in aplastic anemia and death in most of these mice and that this is associated with bone marrow infiltration by antiviral cytotoxic T lymphocytes (CTL) that secrete inflammatory cytokines. We report here that perforin-deficient mice exhibit severe immunopathology in multiple organs that is characterized by infiltration of anti-LCMV CTL that secrete large amounts of gamma interferon (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha). Importantly, this immunopathology is significantly reduced and long-term survival of LCMV infection is increased in perforin-deficient mice expressing LCMV nucleoprotein (NP) in the thymus (and therefore deleting most of their LCMV-NP CTL) compared to the situation in thymus nonexpressors. This is due to the selective reduction of NP-specific CTL responses and their inflammatory-cytokine (IFN-gamma and TNF-alpha) secretion and to a lack of pathogenetically relevant compensatory responses to other viral proteins. Thus, "selective reduction" of the antiviral immune response to only one viral protein can significantly reduce inflammatory immunopathology and might be a therapeutic possibility for certain nonlytic infections. PMID- 10364345 TI - Human herpesvirus 6 open reading frame U83 encodes a functional chemokine. AB - Some viruses including herpesviruses have undergone evolution to benefit viral infection and propagation by pirating and modifying host genes such as chemokine genes. Human herpesvirus 6 (HHV-6), acutely or persistently infects mononuclear cells in vitro. DNA sequence analysis of HHV-6 has revealed that the putative protein encoded by an open reading frame (ORF) of the U83 gene in HHV-6 variant B resembled a human chemokine. We have cloned the U83 gene and analyzed the biological function of this gene. The U83 gene contained an ORF encoding a 113 amino-acid peptide, starting at the first methionine and containing a possible signal peptide and the typical cysteine residues characteristic of the chemokines. Reverse transcription-PCR analysis of mRNA and immunofluorescent antibody testing of infected cells both indicated that the encoded protein was a late protein. The ORF U83 gene fused to the Fc gene was expressed as a fusion protein in COS-7 cells by transfection, and the fusion protein was purified from the supernatant of transfected cells to test its biological function. The purified protein was capable of inducing transient calcium mobilization in THP-1 cells and of chemotactically activating THP-1 cells. These findings suggested that the U83 protein might play an important role in HHV-6 propagation in vivo by activating and trafficking mononuclear cells to sites of viral replication, thus aiding the development of superbly efficient virus production mechanisms. PMID- 10364346 TI - Neutralizing antibodies inhibit axonal spread of herpes simplex virus type 1 to epidermal cells in vitro. AB - The ability of antibodies to interfere with anterograde transmission of herpes simplex virus (HSV) from neuronal axons to the epidermis was investigated in an in vitro model consisting of human fetal dorsal root ganglia innervating autologous skin explants in a dual-chamber tissue culture system. The number and size of viral cytopathic plaques in epidermal cells after axonal transmission from HSV type 1 (HSV-1)-infected dorsal root ganglionic neurons were significantly reduced by addition to the outer chamber of neutralizing polyclonal human sera to HSV-1, of a human recombinant monoclonal group Ib antibody to glycoprotein D (gD), and of rabbit sera to HSV-1 gB and gD but not by rabbit anti gE or anti-gG. A similar pattern of inhibition of direct infection of epidermal cells by these antibodies was observed. High concentrations of the monoclonal anti-gD reduced transmission by 90%. Rabbit anti-gB was not taken up into neurons, and human anti-gD did not influence spread of HSV in the dorsal root ganglia or axonal transport of HSV antigens when applied to individual dissociated neurons. These results suggest that anti-gD and -gB antibodies interfere with axonal spread of HSV-1, possibly by neutralizing HSV during transmission across an intercellular gap between axonal termini and epidermal cells, and thus contribute to control of HSV spread and shedding. Therefore, selected human monoclonal antibodies to protective epitopes might even be effective in preventing epidermis-to-neuron transmission during primary HSV infection, especially neonatal infection. PMID- 10364347 TI - Interaction of peptides with sequences from the Newcastle disease virus fusion protein heptad repeat regions. AB - Typical of many viral fusion proteins, the sequence of the Newcastle disease virus (NDV) fusion protein has several heptad repeat regions. One, HR1, is located just carboxyl terminal to the fusion peptide, while the other, HR2, is located adjacent to the transmembrane domain. The structure and function of a synthetic peptide with a sequence from the region of the NDV HR1 region (amino acids 150 to 173) were characterized. The peptide inhibited fusion with a half maximal concentration of approximately 2 microM; however, inhibition was observed only if the peptide was added prior to protease activation of the fusion protein. This inhibition was virus specific since the peptide had minimal effect on fusion directed by the Sendai virus glycoproteins. To explore the mechanism of action, the potential HR1 peptide interaction with a previously characterized fusion inhibitory peptide with a sequence from the HR2 domain (J. K. Young, R. P. Hicks, G. E. Wright, and T. G. Morrison, Virology 238:291-304, 1997) was characterized. The results demonstrated an interaction between the two peptides both functionally and directly. First, while the individual peptides each inhibit fusion, equimolar mixtures of the two peptides had minimal effect on fusion, suggesting that the two peptides form a complex preventing their interaction with a target protein. Second, an HR2 peptide covalently linked with biotin was found to bind specifically to HR1 peptide in a Western blot. The structure of the HR1 peptide was analyzed by nuclear magnetic resonance spectroscopy and found to be an alpha helix. PMID- 10364348 TI - Colocalization and membrane association of murine hepatitis virus gene 1 products and De novo-synthesized viral RNA in infected cells. AB - Murine hepatitis virus (MHV) gene 1, the 22-kb polymerase (pol) gene, is first translated into a polyprotein and subsequently processed into multiple proteins by viral autoproteases. Genetic complementation analyses suggest that the majority of the gene 1 products are required for viral RNA synthesis. However, there is no physical evidence supporting the association of any of these products with viral RNA synthesis. We have now performed immunofluorescent-staining studies with four polyclonal antisera to localize various MHV-A59 gene 1 products in virus-infected cells. Immunoprecipitation experiments showed that these antisera detected proteins representing the two papain-like proteases and the 3C like protease encoded by open reading frame (ORF) 1a, the putative polymerase (p100) and a p35 encoded by ORF 1b, and their precursors. De novo-synthesized viral RNA was labeled with bromouridine triphosphate in lysolecithin permeabilized MHV-infected cells. Confocal microscopy revealed that all of the viral proteins detected by these antisera colocalized with newly synthesized viral RNA in the cytoplasm, particularly in the perinuclear region of infected cells. Several cysteine and serine protease inhibitors, i.e., E64d, leupeptin, and zinc chloride, inhibited viral RNA synthesis without affecting the localization of viral proteins, suggesting that the processing of the MHV gene 1 polyprotein is tightly associated with viral RNA synthesis. Dual labeling with antibodies specific for cytoplasmic membrane structures showed that MHV gene 1 products and RNA colocalized with the Golgi apparatus in HeLa cells. However, in murine 17CL-1 cells, the viral proteins and viral RNA did not colocalize with the Golgi apparatus but, instead, partially colocalized with the endoplasmic reticulum. Our results provide clear physical evidence that several MHV gene 1 products, including the proteases and the polymerase, are associated with the viral RNA replication-transcription machinery, which may localize to different membrane structures in different cell lines. PMID- 10364349 TI - Replication of murine cytomegalovirus in differentiated macrophages as a determinant of viral pathogenesis. AB - Blood monocytes or tissue macrophages play a pivotal role in the pathogenesis of murine cytomegalovirus (MCMV) infection, providing functions beneficial to both the virus and the host. In vitro and in vivo studies have indicated that differentiated macrophages support MCMV replication, are target cells for MCMV infection within tissues, and harbor latent MCMV DNA. However, this cell type presumably initiates early, antiviral immune responses as well. In addressing this paradoxical role of macrophages, we provide evidence that the proficiency of MCMV replication in macrophages positively correlates with virulence in vivo. An MCMV mutant from which the open reading frames M139, M140, and M141 had been deleted (RV10) was defective in its ability to replicate in macrophages in vitro and was highly attenuated for growth in vivo. However, depletion of splenic macrophages significantly enhanced, rather than deterred, replication of both wild-type (WT) virus and RV10 in the spleen. The ability of RV10 to replicate in intact or macrophage-depleted spleens was independent of cytokine production, as this mutant virus was a poor inducer of cytokines compared to WT virus in both intact organs and macrophage-depleted organs. Macrophages were, however, a major contributor to the production of tumor necrosis factor alpha and gamma interferon in response to WT virus infection. Thus, the data indicate that tissue macrophages serve a net protective role and may function as "filters" in protecting other highly permissive cell types from MCMV infection. The magnitude of virus replication in tissue macrophages may dictate the amount of virus accessible to the other cells. Concomitantly, infection of this cell type initiates the production of antiviral immune responses to guarantee efficient clearance of acute MCMV infection. PMID- 10364350 TI - Cytotoxic T-lymphocyte epitope immunodominance in the control of choroid plexus tumors in simian virus 40 large T antigen transgenic mice. AB - The simian virus 40 (SV40) large tumor antigen (Tag) is a virus-encoded oncoprotein which is the target of a strong cytotoxic T-lymphocyte (CTL) response. Three immunodominant H-2(b)-restricted epitopes, designated epitopes I, II/III, and IV, have been defined. We investigated whether induction of CTLs directed against these Tag epitopes might control Tag-induced tumors in SV11(+) (H-2(b)) mice. SV11(+) mice develop spontaneous tumors of the choroid plexus due to expression of SV40 Tag as a transgene. We demonstrate that SV11(+) mice are functionally tolerant to the immunodominant Tag CTL epitopes. CTLs specific for the H-2Kb-restricted Tag epitope IV were induced in SV11(+) mice following adoptive transfer with unprimed C57BL/6 spleen cells and immunization with recombinant vaccinia viruses expressing either full-length Tag or the H-2Kb restricted epitope IV as a minigene. In addition, irradiation of SV11(+) mice prior to adoptive transfer with unprimed C57BL/6 spleen cells led to the priming of epitope IV-specific CTLs by the endogenous Tag. Induction of epitope IV specific CTLs in SV11(+) mice by either approach correlated with increased life span and control of the choroid plexus tumor progression, indicating that CTLs specific for the immunodominant Tag epitope IV control the progressive growth of spontaneous tumors induced by this DNA virus oncogene in transgenic mice. PMID- 10364352 TI - Expression of the open reading frame 74 (G-protein-coupled receptor) gene of Kaposi's sarcoma (KS)-associated herpesvirus: implications for KS pathogenesis. AB - Kaposi's sarcoma (KS)-associated herpesvirus (KSHV) encodes a G-protein-coupled receptor (GCR) homolog. This protein is a potent, constitutively active signalling molecule that can influence both proliferation and angiogenesis when ectopically expressed in fibroblasts in vitro. Here we have examined the expression of the KSHV GCR gene in virus-infected lymphoid cells and in KS tumors. Our results show that in both situations the gene is expressed primarily during lytic replication; its transcription is unaffected by inhibition of viral DNA synthesis, indicating that it is expressed in the early phases of the lytic program. The major transcript bearing GCR sequences is bicistronic, harboring coding sequences for another viral gene, K14, at its 5' end. Extensive searches for monocistronic GCR mRNAs using nuclease mapping and reverse transcription-PCR failed to detect such species. The 5' end of K14/GCR mRNA maps to nucleotide (nt) 127848, and its poly(A) addition site maps to nt 130546; a 149-nt intron is present in the K14/GCR intergenic region. These results suggest that the KSHV GCR is translated by unconventional mechanisms involving either translational reinitiation, internal ribosomal entry, or leaky ribosomal scanning. The restriction of GCR expression to the lytic cycle has important implications for the potential role(s) of the GCR in KS pathogenesis. PMID- 10364351 TI - Receptor-mediated Moloney murine leukemia virus entry can occur independently of the clathrin-coated-pit-mediated endocytic pathway. AB - To investigate receptor-mediated Moloney murine leukemia virus (MoMuLV) entry, the green fluorescent protein (GFP)-tagged ecotropic receptor designated murine cationic amino acid transporter (MCAT-1) (MCAT-1-GFP) was constructed and expressed in 293 cells (293/MCAT-1-GFP). 293/MCAT-1-GFP cells displayed green fluorescence primarily at the cell membrane and supported wild-type levels of MoMuLV vector binding and transduction. Using immunofluorescence labeling and confocal microscopy, it was demonstrated that the surface envelope protein (SU) gp70 of MoMuLV virions began to appear inside cells 5 min after virus binding and was colocalized with MCAT-1-GFP. However, clathrin was not colocalized with MCAT 1-GFP, suggesting that MoMuLV entry, mediated by MCAT-1, does not involve clathrin. Double immunofluorescence labeling of SU and clathrin in 293 cells expressing untagged receptor (293/MCAT-1) gave the same results, i.e., SU and clathrin did not colocalize. In addition, we examined the transduction ability of MoMuLV vector on HeLa cells overexpressing the dominant-negative GTPase mutant of dynamin (K44A). HeLa cells overexpressing mutant dynamin have a severe block in endocytosis by the clathrin-coated-pit pathway. No significant titer difference was observed when MoMuLV vector was tranduced into HeLa cells overexpressing either wild-type or mutant dynamin, while the transduction ability of vesicular stomatitis virus glycoprotein pseudotyped vector into HeLa cells overexpressing mutant dynamin was decreased significantly. Taken together, these data suggest that MoMuLV entry does not occur through the clathrin-coated-pit-mediated endocytic pathway. PMID- 10364353 TI - Hepatitis A virus capsid protein VP1 has a heterogeneous C terminus. AB - Hepatitis A virus (HAV) encodes a single polyprotein which is posttranslationally processed into the functional structural and nonstructural proteins. Only one protease, viral protease 3C, has been implicated in the nine protein scissions. Processing of the capsid protein precursor region generates a unique intermediate, PX (VP1-2A), which accumulates in infected cells and is assumed to serve as precursor to VP1 found in virions, although the details of this reaction have not been determined. Coexpression in transfected cells of a variety of P1 precursor proteins with viral protease 3C demonstrated efficient production of PX, as well as VP0 and VP3; however, no mature VP1 protein was detected. To identify the C-terminal amino acid residue of HAV VP1, we performed peptide sequence analysis by protease-catalyzed [18O]H2O incorporation followed by liquid chromatography ion-trap microspray tandem mass spectrometry of HAV VP1 isolated from purified virions. Two different cell culture-adapted isolates of HAV, strains HM175pE and HM175p35, were used for these analyses. VP1 preparations from both virus isolates contained heterogeneous C termini. The predominant C-terminal amino acid in both virus preparations was VP1-Ser274, which is located N terminal to a methionine residue in VP1-2A. In addition, the analysis of HM175pE recovered smaller amounts of amino acids VP1-Glu273 and VP1-Thr272. In the case of HM175p35, which contains valine at amino acid position VP1-273, VP1-Thr272 was found in addition to VP1-Ser274. The data suggest that HAV 3C is not the protease responsible for generation of the VP1 C terminus. We propose the involvement of host cell protease(s) in the production of HAV VP1. PMID- 10364354 TI - Ebola virus can be effectively neutralized by antibody produced in natural human infection. AB - The activity of antibodies against filoviruses is poorly understood but has important consequences for vaccine design and passive prophylaxis. To investigate this activity, a panel of recombinant human monoclonal antibodies to Ebola virus antigens was isolated from phage display libraries constructed from RNA from donors who recovered from infection in the 1995 Ebola virus outbreak in Kikwit, Democratic Republic of Congo. Antibodies reactive with nucleoprotein (NP), envelope glycoprotein (GP), and secreted envelope glycoprotein (sGP) were characterized by immunofluorescence and radioimmunoprecipitation assays. Four antibodies reacting strongly with sGP and weakly with GP and two antibodies reacting with NP were not neutralizing. An antibody specific for GP neutralized Ebola virus to 50% at 0.4 microgram/ml as the recombinant Fab fragment and to 50% at 0.3 microgram/ml (90% at 2.6 microgram/ml) as the corresponding whole immunoglobulin G1 molecule. The studies indicate that neutralizing antibodies are produced in infection by Ebola virus although probably at a relatively low frequency. The neutralizing antibody may be useful in vaccine design and as a prophylactic agent against Ebola virus infection. PMID- 10364355 TI - Induction of adult T-cell leukemia-like lymphoproliferative disease and its inhibition by adoptive immunotherapy in T-cell-deficient nude rats inoculated with syngeneic human T-cell leukemia virus type 1-immortalized cells. AB - Human T-cell leukemia virus type 1 (HTLV-1) has been shown to be the etiologic agent of adult T-cell leukemia (ATL), but the in vivo mechanism by which the virus causes the malignant transformation is largely unknown. In order to investigate the mechanisms of HTLV-1 leukemogenesis, we developed a rat model system in which ATL-like disease was reproducibly observed, following inoculation of various rat HTLV-1-immortalized cell lines. When previously established cell lines, F344-S1 and TARS-1, but not TART-1 or W7TM-1, were inoculated, systemic multiple tumor development was observed in adult nude (nu/nu) rats. FPM1 cells, newly established from a heterozygous (nu/+) rat syngeneic to nu/nu rats, caused transient tumors only at the injection site in adult nu/nu rats, but could progressively grow in newborn nu/nu rats and metastasize in lymph nodes. The derivative cell line (FPM1-V1AX) serially passed through newborn nu/nu rats acquired the potency to grow in adult nu/nu rats. These results indicated that only some with additional changes but not all of the in vitro HTLV-1-immortalized cell lines possessed in vivo tumorigenicity. Using the syngeneic system, we further showed the inhibition of tumor development by transferring splenic T cells from immunized rats, suggesting the involvement of T cells in the regression of tumors. This novel and reproducible nude rat model of human ATL would be useful for investigation of leukemogenesis and antitumor immune responses in HTLV-1 infection. PMID- 10364356 TI - Isolation and molecular characterization of a poliovirus type 1 mutant that replicates in the spinal cords of mice. AB - The Mahoney strain of poliovirus type 1 (OM) is generally unable to cause paralysis in mice. We isolated a mouse-adapted mutant, PV1/OM-SA (SA), from the spinal cord of a mouse that had been intracerebrally inoculated with OM. SA showed mouse neurovirulence only with intraspinal inoculation, and the infected mice developed a flaccid paralysis, which was indistinguishable from that observed in poliovirus-sensitive transgenic mice inoculated with OM. SA antigens were detected in neurons of the spinal cords of the infected mice. Nucleotide (nt) sequence analysis revealed 9 nt changes on the SA genome, resulting in three amino acid (a.a.) substitutions, i.e., one each in the capsid proteins VP4 and VP1 and in the noncapsid protein 2C. To identify the key mutation site(s) for the mouse neurovirulence, virus recombinants between OM and SA were constructed by using infectious cDNA clones of these two viruses and tested for their mouse neurovirulence after inoculation via an intraspinal route. The results indicated that a mutation at nt 928 (replacement of A with G), resulting in a substitution of Met for Ile at a.a. 62 within VP4, was responsible for conferring the mouse neurovirulence phenotype of the mutant SA. The mutation in VP4 may render the virus accessible to a molecule that acts as a virus receptor and is located on the surfaces of neurons of the mouse spinal cord. This molecule appears not to be expressed in the mouse brain. PMID- 10364357 TI - Generation of an adenovirus vector lacking E1, e2a, E3, and all of E4 except open reading frame 3. AB - Toxicity and immunity associated with adenovirus backbone gene expression is an important hurdle to overcome for successful gene therapy. Recent efforts to improve adenovirus vectors for in vivo use have focused on the sequential deletion of essential early genes. Adenovirus vectors have been constructed with the E1 gene deleted and with this deletion in combination with an E2a, E2b, or E4 deletion. We report here a novel vector (Av4orf3nBg) lacking E1, E2a, and all of E4 except open reading frame 3 (ORF3) and expressing a beta-galactosidase reporter gene. This vector was generated by transfection of a plasmid carrying the full-length vector sequence into A30.S8 cells that express E1 and E2a but not E4. Production was subsequently performed in an E1-, E2a-, and E4-complementing cell line. We demonstrated with C57BL/6 mice that the Av4orf3nBg vector effected gene transfer with an efficiency comparable to that of the Av3nBg (wild-type E4) vector but that the former exhibited a higher level of beta-galactosidase expression. This observation suggests that E4 ORF3 alone is able to enhance RNA levels from the beta-galactosidase gene when the Rous sarcoma virus promoter is used to drive transgene expression in the mouse liver. In addition, we observed less liver toxicity in mice injected with the Av4orf3nBg vector than those injected with the Av3nBg vector at a comparable DNA copy number per cell. This study suggests that the additional deletion of E4 in an E1 and E2a deletion background may be beneficial in decreasing immunogenicity and improving safety and toxicity profiles, as well as increasing transgene capacity and expression for liver-directed gene therapy. PMID- 10364358 TI - Fiber swap between adenovirus subgroups B and C alters intracellular trafficking of adenovirus gene transfer vectors. AB - Following receptor binding and internalization, intracellular trafficking of adenovirus (Ad) among subgroups B and C is different, with significant amounts of Ad serotype 7 (Ad7) (subgroup B) virions found in cytoplasm during the initial hours of infection while Ad5 (subgroup C) virions rapidly translocate to the nucleus. To evaluate the role of the fiber in these differences, we examined intracellular trafficking of Ad5, Ad7, and Ad5f7 (a chimeric vector composed of the Ad5 capsid with the fiber replaced by the Ad7 fiber) by conjugating Ad capsids directly with Cy3 fluorescent dye, permitting the trafficking of the capsids to be examined by fluorescence microscopy. The human lung carcinoma cell line A549 was infected with Cy3-conjugated viruses for 10 min followed by a 1-h incubation. Ad5 virions rapidly translocated to the nucleus (within 1 h of infection), while Ad7 virions were widely distributed in the cytoplasm at the same time point. Interestingly, chimeric Ad5f7 virions behaved similarly to Ad7 but not Ad5. In this regard, the percentages of nuclear localization of Ad5, Ad7, and Ad5f7 at 1 h following infection were 72% +/- 4%, 32% +/- 6%, and 38% +/- 2%, respectively. Consistent with these observations, fluorescence in situ hybridization demonstrated that most of the Ad5 DNA was detected at the nucleus after 1 h, but at the same time point, DNA of Ad7 and Ad5f7 was distributed in both the nucleus and cytoplasm. Quantification of the kinetics of Ad genomic DNA delivery to the nucleus using a fluorogenic probe-based PCR assay (TaqMan PCR) demonstrated that the percentages of nuclear association of Ad5 DNA and Ad5f7 DNA at 1 h postinfection were 80% +/- 13% and 43% +/- 1%, respectively. Although it has been generally accepted that Ad fiber protein mediates attachment of virions to cells and that fibers dissociate during endocytic uptake, these data suggest that in addition to mediating binding to the cell surface, fiber likely modulates intracellular trafficking as well. PMID- 10364359 TI - Poliovirus induces apoptosis in the mouse central nervous system. AB - Poliovirus (PV) is the etiological agent of human paralytic poliomyelitis. Paralysis results from the destruction of motoneurons, a consequence of PV replication. However, the PV-induced process leading to the death of motoneurons is not well known. We investigated whether PV-induced central nervous system (CNS) injury is associated with apoptosis by using mice as animal models. Transgenic mice expressing the human PV receptor were infected intracerebrally with either the neurovirulent PV-1 Mahoney strain or a paralytogenic dose of the attenuated PV-1 Sabin strain. Nontransgenic mice were infected with a mouse adapted PV-1 Mahoney mutant. DNA fragmentation was demonstrated in CNS tissue from paralyzed mice by visualization of DNA oligonucleosomal laddering and by enzyme-linked immunosorbent assay. Viral antigens and DNA fragmentation detected by the in situ terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling technique were colocalized in neurons of spinal cords from paralyzed mice. In addition, morphological changes characteristic of cells undergoing apoptosis were observed in motoneurons by electron microscopy. Thus, we show that PV multiplication and CNS injury during paralytic poliomyelitis are associated with apoptosis. PMID- 10364360 TI - The novel structural protein of human cytomegalovirus, pUL25, is localized in the viral tegument. AB - Human cytomegalovirus UL25 codes for a structural phosphoprotein of 85 kDa (C. J. Baldick and T. Shenk, J. Virol. 70:6097-6105, 1996; M. C. Battista et al., J. Virol. 73:3800-3809, 1999). In this study we analyzed the intracellular and intraviral localization of pUL25 by confocal and immunoelectron microscopy and found that pUL25 is a component of the viral tegument and the dense body matrix, acquired during the late cytoplasmic phase of virus maturation. PMID- 10364361 TI - Association of JC virus large T antigen with myelin basic protein transcription factor (MEF-1/Puralpha) in hypomyelinated brains of mice transgenically expressing T antigen. AB - Progressive multifocal leukoencephalopathy (PML) is a fatal demyelinating disease caused by cytolytic destruction of oligodendrocytes, the myelin-producing cells of the central nervous system, by the human neurotropic JC virus (JCV). The early protein of JCV, T antigen, which is produced at the early stage of infection, is important for orchestrating the events leading to viral lytic infection and cytolytic destruction of oligodendrocytes. Results from transgenic mouse studies, however, have revealed that, in the absence of lytic infection, this protein can induce brain hypomyelination and suppression of myelin gene expression. Since expression of the gene encoding myelin basic protein, the major component of myelin, can be regulated by a DNA-binding transcription factor, MEF-1/Puralpha, (Puralpha), we have examined the level of this protein in transgenic mouse brains. Results from immunoprecipitation and Western blots showed that while there was no drastic decrease in the level of MEF-1/Puralpha in transgenic mouse brains, JCV T antigen was found in a complex with MEF-1/Puralpha. Immunohistological studies revealed abnormal oligodendrocytes in white matter, where MEF-1/Puralpha and T antigen were detected. Furthermore, immunogold electron microscopic studies revealed that Puralpha and T antigen are colocalized in the nucleus of the oligodendrocytes and in hippocampal neurons. Interestingly, results from cell culture studies revealed that incubation of oligodendrocytes with JCV led to a drastic decrease in the level of MEF-1/Puralpha protein. These observations provide insight into the molecular pathogenesis of PML and support a model for a dual effect of JCV on inducing hypomyelination by (i) affecting myelin gene expression via interaction of JCV T antigen and the myelin gene transcription factor, MEF-1/Puralpha, and (ii) causing a decline in the level of the host regulatory proteins, including MEF-1/Puralpha, which are involved in myelin gene expression. PMID- 10364362 TI - Transduction of well-differentiated airway epithelium by recombinant adeno associated virus is limited by vector entry. AB - The limitations of adeno-associated virus (AAV)-mediated vectors for lung directed gene transfer were investigated by using differentiated human respiratory epithelium in air-liquid interface cultures. Transduction efficiency was high in undifferentiated cells and was enhanced in well-differentiated cells after basolateral application of the vector or after apical application following disruption of tight junctions or pretreatment of the cultures with glycosidases. These results indicate that transduction of airway epithelia by AAV vectors is limited by entry and reinforce the importance of a physical barrier on the airway surface. PMID- 10364363 TI - Role of the membrane-proximal domain in the initial stages of human immunodeficiency virus type 1 envelope glycoprotein-mediated membrane fusion. AB - We have examined mutations in the ectodomain of the human immunodeficiency virus type 1 transmembrane glycoprotein gp41 within a region immediately adjacent to the membrane-spanning domain for their effect on the outcome of the fusion cascade. Using the recently developed three-color assay (I. Munoz-Barroso, S. Durell, K. Sakaguchi, E. Appella, and R. Blumenthal, J. Cell Biol. 140:315-323, 1998), we have assessed the ability of the mutant gp41s to transfer lipid and small solutes from susceptible target cells to the gp120-gp41-expressing cells. The results were compared with the syncytium-inducing capabilities of these gp41 mutants. Two mutant proteins were incapable of mediating both dye transfer and syncytium formation. Two mutant proteins mediated dye transfer but were less effective at inducing syncytium formation than was wild-type gp41. The most interesting mutant proteins were those that were not capable of inducing syncytium formation but still mediated dye transfer, indicating that the fusion cascade was blocked beyond the stage of small fusion pore formation. Fusion mediated by the mutant gp41s was inhibited by the peptides DP178 and C34. PMID- 10364364 TI - The GDVII strain of Theiler's virus spreads via axonal transport. AB - Following intracerebral inoculation, the DA strain of Theiler's virus sequentially infects neurons in the gray matter and glial cells in the white matter of the spinal cord. It persists in the latter throughout the life of the animal. Several observations suggest that the virus spreads from the gray to the white matter by axonal transport. In contrast, the neurovirulent GDVII strain causes a fatal encephalitis with lytic infection of neurons. It does not infect the white matter of the spinal cord efficiently and does not persist in survivors. The inability of this virus to infect the white matter could be due to a defect in axonal transport. Using footpad inoculations, we showed that the GDVII strain is, in fact, transported in axons. Transport was prevented by sectioning the sciatic nerve. The kinetics of transport and experiments using colchicine suggested that the virus uses microtubule-associated fast axonal transport. Our results show that a cardiovirus can spread by fast axonal transport and suggest that the inability of the GDVII strain to infect the white matter is not due to a defect in axonal transport. PMID- 10364366 TI - Dengue virus type 1 nonstructural glycoprotein NS1 is secreted from mammalian cells as a soluble hexamer in a glycosylation-dependent fashion. AB - Nonstructural glycoprotein NS1, specified by dengue virus type 1 (Den-1), is secreted from infected green monkey kidney (Vero) cells in a major soluble form characterized by biochemical and biophysical means as a unique hexameric species. This noncovalently bound oligomer is formed by three dimeric subunits and has a molecular mass of 310 kDa and a Stokes radius of 64.4 A. During protein export, one of the two oligosaccharides of NS1 is processed into an endo-beta-N acetylglucosaminidase F-resistant complex-type sugar while the other remains of the polymannose type, protected in the dimeric subunit from the action of maturation enzymes. Complete processing of the complex-type sugar appears to be required for efficient release of soluble NS1 into the culture fluid of infected cells, as suggested by the repressive effects of the N-glycan processing inhibitors swainsonine and deoxymannojyrimicin. These results, together with observations related to the absence of secretion of NS1 from Den-infected insect cells, suggest that maturation and secretion of hexameric NS1 depend on the glycosylation status of the host cell. PMID- 10364367 TI - Interaction of eukaryotic initiation factor eIF4B with the internal ribosome entry site of foot-and-mouth disease virus is independent of the polypyrimidine tract-binding protein. AB - Eukaryotic translation initiation factor 4B (eIF4B) binds directly to the internal ribosome entry site (IRES) of foot-and-mouth disease virus (FMDV). Mutations in all three subdomains of the IRES stem-loop 4 reduce binding of eIF4B and translation efficiency in parallel, indicating that eIF4B is functionally involved in FMDV translation initiation. In reticulocyte lysate devoid of polypyrimidine tract-binding protein (PTB), eIF4B still bound well to the wild type IRES, even after removal of the major PTB-binding site. In conclusion, the interaction of eIF4B with the FMDV IRES is essential for IRES function but independent of PTB. PMID- 10364365 TI - Use of real-time PCR and molecular beacons to detect virus replication in human immunodeficiency virus type 1-infected individuals on prolonged effective antiretroviral therapy. AB - We have designed a novel, precise, and sensitive assay to measure unspliced (US) human immunodeficiency virus type 1 (HIV-1) mRNA in peripheral blood mononuclear cells of HIV-1-infected individuals by using real-time PCR and molecular beacons. Individuals were classified as either well suppressed (WS) or partially suppressed, based on longitudinal measurements of plasma HIV-1 RNA. The proportion of individuals with US mRNA undetectable over time was significantly higher among WS individuals; however, 30% of WS subjects still had detectable US mRNA after 24 months of effective antiviral therapy. PMID- 10364368 TI - Retroviral vectors pseudotyped with lymphocytic choriomeningitis virus. AB - Pseudotyping can improve retroviral vector stability and transduction efficiency. Here, we describe a novel pseudotype of murine leukemia virus packaged with lymphocytic choriomeningitis virus (LCMV). This pseudotype was stable during ultracentrifugation and infected several cell lines from different species. Moreover, LCMV glycoproteins were not cell toxic. PMID- 10364369 TI - Replication and pathogenicity of primer binding site mutants of SL3-3 murine leukemia viruses. AB - Retroviral reverse transcription is primed by a cellular tRNA molecule annealed to an 18-bp primer binding site sequence. The sequence of the primer binding site coincides with that of a negatively acting cis element that mediates transcriptional silencing of murine leukemia virus (MLV) in undifferentiated embryonic cells. In this study we test whether SL3-3 MLV can replicate stably using tRNA primers other than the cognate tRNAPro and analyze the effect of altering the primer binding site sequence to match the 3' end of tRNA1Gln, tRNA3Lys, or tRNA1,2Arg in a mouse pathogenicity model. Contrary to findings from cell culture studies of primer binding site-modified human immunodeficiency virus type 1 and avian retroviruses, our findings were that SL3-3 MLV may stably and efficiently replicate with tRNA primers other than tRNAPro. Although lymphoma induction of the SL3-3 Lys3 mutant was significantly delayed relative to that of the wild-type virus, molecular tumor analysis indicated that all the primer binding site-modified viruses induce T-cell lymphomas similar to those induced by the wild-type virus in terms of frequencies of genomic rearrangements within the T-cell receptor beta-chain, the immunoglobulin kappa light chain, and the c-myc locus. Whereas none of the mutants were found to revert to tRNAPro primer utilization, in two tumors resulting from the injection of the SL3-3 Lys3 mutant the primer binding site was altered to match that of a new primer species, tRNA1,2Lys. In addition, recombination with endogenous viruses resulting in the generation of recombinant viruses carrying a glutamine primer binding site was detected in the majority of the tumors induced by the SL3-3 Lys3 mutant as well as in two tumors induced by wild-type SL3-3 and the SL3-3 Arg1,2 mutant. PMID- 10364370 TI - Transient association of calnexin and calreticulin with newly synthesized G1 and G2 glycoproteins of uukuniemi virus (family Bunyaviridae). AB - The membrane glycoproteins G1 and G2 of Uukuniemi virus, a member of the Bunyaviridae family, are cotranslationally cleaved from a common precursor in the endoplasmic reticulum (ER). Here, we show that newly made G1 and G2 associate transiently with calnexin and calreticulin, two lectins involved in glycoprotein folding in the ER. Stable complexes between G1-G2 and calnexin or calreticulin could be immunoprecipitated after solubilization of virus-infected BHK21 cells with the detergents digitonin or Triton X-100. In addition, G1-G2-calnexin complexes could be recovered after solubilization with CHAPS (3-[(3 cholamidopropyl)-dimethylammonio]-1-propane sulfonate), while G1-G2-calreticulin complexes were not readily detected by using this detergent. Only endoglycosidase H-sensitive forms of G1 were found complexed with calnexin. Pulse-chase experiments showed that G1 and G2 associated with both chaperones transiently for up to 120 min. Sequential immunoprecipitations with anticalreticulin and anticalnexin antisera indicated that about 50% of newly synthesized G1 and G2 was associated with either calnexin or calreticulin. Our previous results have shown that newly synthesized G1 and G2 transiently interact also with the ER chaperone BiP and with protein disulfide isomerase (R. Persson and R. F. Pettersson, J. Cell Biol. 112:257-266, 1991). Taking all of this into consideration, we conclude that the folding of G1 and G2 in the ER is catalyzed by at least four different folding factors. PMID- 10364371 TI - Insertion of a new transcriptional unit into the genome of mouse hepatitis virus. AB - The subgenomic mRNAs of the coronavirus mouse hepatitis virus (MHV) are composed of a leader sequence, identical to the 5' 70 nucleotides of the genome, joined at distant downstream sites to a stretch of sequence that is identical to the 3' end of the genome. The points of fusion occur at intergenic sequences (IGSs), loci on the genome that contain a tract of sequence homologous to the 3' end of the leader RNA. We have constructed a mutant of MHV-A59 containing an extra IGS inserted into the genome immediately downstream of the 3'-most gene, that encoding the nucleocapsid (N) protein. We show that in cells infected with the mutant, there is synthesis of an additional leader-containing subgenomic RNA of the predicted size. Our study demonstrates that (i) an IGS can be a sufficient cis-acting element to dictate MHV transcription, (ii) the relative efficiency of an IGS must be influenced by factors other than the nucleotides immediately adjacent to the 5'AAUCUAAAC3' core consensus sequence or its position relative to the 3' end of the genome, (iii) a downstream IGS can exert a polar attenuating effect on upstream IGSs, and (iv) unknown factors prevent the insertion of large exogenous elements between the N gene and the 3' untranslated region of MHV. These results confirm and extend conclusions previously derived from the analysis of defective interfering RNAs. PMID- 10364373 TI - Frequency and stability of chromosomal integration of adenovirus vectors. AB - One of the limitations of adenovirus vectors is the lack of machinery necessary for their integration into host chromosomes, resulting in short-term gene expression in dividing cells. We analyzed frequencies of integration and persistence of gene expression from integrated adenovirus vectors. Both E1 substituted and helper-dependent adenovirus vectors achieved similar integration efficiencies of approximately 10(-3) to 10(-5) per cell, with the helper dependent vector showing slightly higher efficiencies. In stable cell pools, gene expression of the integrated vector persisted for at least 50 cell divisions without selection. However, some stable cell clones showed changes in gene expression, which were accompanied by structural changes in the integrated vector DNA. PMID- 10364372 TI - Identification of human herpesvirus 8-specific cytotoxic T-cell responses. AB - Human herpesvirus 8 (HHV-8) (or Kaposi's sarcoma-associated herpesvirus) is implicated in the etiopathogenesis of Kaposi's sarcoma (KS) and certain lymphoproliferations. The introduction of more effective therapies to treat human immunodeficiency virus infection has led to a decline in the incidence of KS and also in the resolution of KS in those already affected. This suggests that cellular immune responses including cytotoxic T lymphocytes (CTLs) could play a vital role in the control of HHV-8 infection and in KS pathogenesis. Here we elucidate HLA class I-restricted, HHV-8-specific cellular immune responses that could be important in the control of HHV-8 infection and subsequent tumor development. We show the presence of CTLs against HHV-8 latent (K12), lytic (K8.1), and highly variable (K1) proteins in infected individuals. PMID- 10364374 TI - Deletion mutagenesis within the dimerization initiation site of human immunodeficiency virus type 1 results in delayed processing of the p2 peptide from precursor proteins. AB - Previous work has shown that deletions of genomic segments at nucleotide (nt) positions +238 to +253, i.e., construct BH10-LD3, or nt positions +261 to +274, i.e., construct BH10-LD4, within the human immunodeficiency virus type 1 (HIV-1) dimerization initiation site (DIS) destroyed DIS secondary structure and dramatically reduced viral replication capacity. Surprisingly, two point mutations located within the viral peptide 2 (p2) and nucleocapsid (NC) protein termed MP2 and MNC, respectively, were able to compensate for this defect. Since the MP2 mutation involves an amino acid substitution near the cleavage site between p2 and NC, we investigated the effects of the above-mentioned deletions on the processing of Gag proteins. Immunoprecipitation assays performed with monoclonal antibodies against viral capsid (CA) (p24) protein showed that p2 was cleaved from CA with less efficiency in viruses that contained the LD3 and LD4 deletions than in wild-type viruses. The presence of the two compensatory mutations, MP2 and MNC, increased the efficiency of the cleavage of p2 from CA, but neither mutation alone had this effect or was sufficient to compensate for the observed impairment in infectiousness. A virus that contained both of the above-mentioned deletions within the DIS was also impaired in regard to processing and infectiousness, and it could likewise be compensated by the MP2 and MNC point mutations. These results suggest that the DIS region of HIV-1 RNA plays an important role in the processing of Gag proteins. PMID- 10364375 TI - Simian immunodeficiency virus and human immunodeficiency virus type 1 nef proteins show distinct patterns and mechanisms of Src kinase activation. AB - The nef gene from human and simian immunodeficiency viruses (HIV and SIV) regulates cell function and viral replication, possibly through binding of the nef product to cellular proteins, including Src family tyrosine kinases. We show here that the Nef protein encoded by SIVmac239 interacts with and also activates the human Src kinases Lck and Hck. This is in direct contrast to the inhibitory effect of HIV type 1 (HIV-1) Nef on Lck catalytic activity. Unexpectedly, however, the interaction of SIV Nef with human Lck or Hck is not mediated via its consensus proline motif, which is known to mediate HIV-1 Nef binding to Src homology 3 (SH3) domains, and various experimental analyses failed to show significant interaction of SIV Nef with the SH3 domain of either kinase. Instead, SIV Nef can bind Lck and Hck SH2 domains, and its N-terminal 50 amino acid residues are sufficient for Src kinase binding and activation. Our results provide evidence for multiple mechanisms by which Nef binds to and regulates Src kinases. PMID- 10364376 TI - Persistent infection of rhesus macaques by the rev-independent Nef(-) simian immunodeficiency virus SIVmac239: replication kinetics and genomic stability. AB - We generated previously a Nef(-), replication-competent clone of SIVmac239 in which the Rev protein and the Rev-responsive element were replaced by the constitutive transport element (CTE) of simian retrovirus type 1 (A. S. von Gegerfelt and B. K. Felber, Virology 232:291-299, 1997). In the present report, we show that this virus was able to infect and replicate in rhesus macaques. The Rev-independent Nef(-) simian immunodeficiency virus induced a persistent humoral immune response in all monkeys, although viral loads were very low. Upon propagation in the monkeys, the genotype remained stable and the virus retained its in vitro growth characteristics. The infected monkeys showed normal hematological values and no signs of disease at more than 18 months post-virus exposure. Therefore, replacement of the essential Rev regulation by the CTE generated a virus variant that retained its replicative capacity both in vitro and in vivo, albeit at low levels. PMID- 10364377 TI - Human papillomavirus type 16 E7 oncoprotein expressed in peripheral epithelium tolerizes E7-directed cytotoxic T-lymphocyte precursors restricted through human (and mouse) major histocompatibility complex class I alleles. AB - Mice which coexpress human papillomavirus type 16 E7 and HLA A2.1 in peripheral squamous epithelium and thymic cortical epithelium are tolerant at the cytotoxic T-lymphocyte (CTL) level to E7 epitopes restricted through HLA A*0201 and H-2(b) (T. Doan, M. Chambers, M. Street, G. J. Fernando, K. Herd, P. Lambert, and R. Tindle, Virology 244:352-364, 1998). Here we used bone marrow-reconstituted radiation chimeras to distinguish whether E7-directed CTL tolerance was mediated peripherally by E7 expression in skin or centrally by E7 expression in thymus. In chimeric mice expressing E7 in skin and reconstituted with E7-naive bone marrow and E7-naive thymus, CTL responses to vaccine-administered E7 epitopes were not restored, i.e. , the mice remained tolerant. In contrast, chimeric mice not expressing E7 in skin and reconstituted with E7-naive bone marrow and E7 expressing thymus had full E7-directed CTL responses. These results demonstrate that E7 protein expression in peripheral squamous epithelium is sufficient to tolerize the E7-directed CTL precursor repertoire. The data have implications for E7-mediated tumorigenesis and for the development of E7-based immunotherapeutic strategies, since peripheral immunological tolerance of tumor-associated antigens may create a barrier to effective immunotherapy. PMID- 10364378 TI - Contributions of viral splice sites and cis-regulatory elements to lentivirus vector function. AB - The mobile transgene constructs of most human immunodeficiency virus (HIV)-based lentivirus vectors currently in use contain viral long terminal repeats, a 5' untranslated region, gag sequences, and env sequences that include the Rev responsive element (RRE). In this study, we examined the possibility of deleting HIV splice sites and gag and env sequences from an HIV type 1 recombinant vector established in our laboratory as part of our ongoing efforts to improve this vector system. Mutations in the major splice donor site (SD) markedly reduced viral RNA expression but had little effect on vector titer. Deletion of gag or env sequences, excluding RRE, led to a moderate reduction in vector titer. Interestingly, deletion of RRE slightly reduced viral RNA expression but markedly impaired vector function. Combined deletions of RRE, gag (except for the first 40 nucleotides), env, and the SD mutation resulted in a twofold increase in cytoplasmic viral RNA expression and a recovery of vector efficiency to approximately 50% of the wild-type level. This increase in cytoplasmic RNA levels is likely to be due, at least in part, to effects of the TE671 host cells, a human rhabdomyosarcoma cell line used for vector production in our system, on the cytoplasmic distribution of spliced and unspliced viral RNA. These results show that optimal lentivirus vector function can be maintained in the absence of multiple essential viral elements. PMID- 10364379 TI - A rhesus macaque rhadinovirus related to Kaposi's sarcoma-associated herpesvirus/human herpesvirus 8 encodes a functional homologue of interleukin-6. AB - The rhesus rhadinovirus strain 17577 (RRV strain 17577) genome is essentially colinear with human herpesvirus 8 (HHV8)/Kaposi's sarcoma-associated herpesvirus (KSHV) and encodes several analogous open reading frames (ORFs), including the homologue of cellular interleukin-6 (IL-6). To determine if the RRV IL-6-like ORF (RvIL-6) is biologically functional, it was expressed either transiently in COS-1 cells or purified from bacteria as a glutathione S-transferase (GST)-RvIL-6 fusion and analyzed by IL-6 bioassays. Utilizing the IL-6-dependent B9 cell line, we found that both forms of RvIL-6 supported cell proliferation in a dose dependent manner. Moreover, antibodies specific to the IL-6 receptor (IL-6R) or the gp130 subunit were capable of blocking the stimulatory effects of RvIL-6. Reciprocal titrations of GST-RvIL-6 against human recombinant IL-6 produced a more-than-additive stimulatory effect, suggesting that RvIL-6 does not inhibit but may instead potentiate normal cellular IL-6 signaling to B cells. These results demonstrate that RRV encodes an accessory protein with IL-6-like activity. PMID- 10364380 TI - The subgenus-specific C-terminal region of protein IX is located on the surface of the adenovirus capsid. AB - We have investigated the antigenicity of the C- and N-terminal halves of pIX of human adenovirus types 2 and 3 (Ad2 and Ad3) as well as their orientations in virions. We found that only the C-terminal halves of Ad2 pIX and Ad3 pIX reacted in a subgenus-specific manner by enzyme-linked immunosorbent assay and immunoblot analysis. Based on immunoelectron microscopy experiments, pIX in viral capsids appears to be positioned such that the C-terminal part of pIX constitutes the surface domain whereas the N terminus of the protein makes up the internal domain in icosahedral Ad capsids. PMID- 10364381 TI - Common neutralization epitope in minor capsid protein L2 of human papillomavirus types 16 and 6. AB - Studies of virus neutralization by antibody are a prerequisite for development of a prophylactic vaccine strategy against human papillomaviruses (HPVs). Using HPV16 and -6 pseudovirions capable of inducing beta-galactosidase in infected monkey COS-1 cells, we examined the neutralizing activity of mouse monoclonal antibodies (MAbs) that recognize surface epitopes in HPV16 minor capsid protein L2. Two MAbs binding to a synthetic peptide with the HPV16 L2 sequence of amino acids (aa) 108 to 120 were found to inhibit pseudoinfections with HPV16 as well as HPV6. Antisera raised by immunizing BALB/c mice with the synthetic peptide had a cross-neutralizing activity similar to that of the MAb. The data indicate that HPV16 and -6 have a common cross-neutralization epitope (located within aa 108 to 120 of L2 in HPV16), suggesting that this epitope may be shared by other genital HPVs. PMID- 10364382 TI - Reappearance of founder virus sequence in human immunodeficiency virus type 1 infected patients. AB - Different patterns of temporal evolution in human immunodeficiency virus type 1 V3 and p17 regions are described for eight patients studied during the first years following primary infection. In samples from three patients, a rapid replacement of the major sequence occurred but the original sequence reappeared later simultaneously with clinical deterioration and increased plasma viral load. PMID- 10364383 TI - Identification of biased amino acid substitution patterns in human immunodeficiency virus type 1 isolates from patients treated with protease inhibitors. AB - Human immunodeficiency virus type 1 (HIV-1) amino acid substitutions observed during antiretroviral drug therapy may be caused by drug selection, non-drug related evolution, or sampling error introduced by the sequencing process. We analyzed HIV-1 sequences from 371 untreated patients and from 178 patients receiving a single protease inhibitor. Amino acid substitution patterns during treatment were compared with inferred substitution patterns arising evolutionarily without treatment. Our results suggest that most treatment associated amino acid substitutions are caused by selective drug pressure, including substitutions not previously associated with drug resistance. PMID- 10364384 TI - Identification of phosphorylation sites within the herpes simplex virus tegument protein VP22. AB - The herpes simplex virus protein VP22 is a major phosphoprotein of infected cells. In this study, we identify two serine phosphorylation sites within VP22 and show that the N-terminal site is a substrate for casein kinase II, while the extreme C-terminal site is a substrate for another, as yet unidentified, cellular kinase. Furthermore, we show that a mutant of VP22 which has both sites altered is unable to incorporate phosphate in vivo, confirming that there are no other phosphorylation sites within VP22. PMID- 10364386 TI - Fructan: more than a reserve carbohydrate? PMID- 10364387 TI - The sensitivity of barley aleurone tissue to gibberellin is heterogeneous and may Be spatially determined AB - In cereals, gibberellin (GA) enhances the synthesis and secretion of hydrolytic enzymes from aleurone cells. These enzymes then mobilize the endosperm storage reserves that fuel germination. The dose-response curve of aleurone protoplasts to GA extends over a range of concentrations from 10(-11) to more than 10(-6) M. One hypothesis is that subpopulations of cells have different sensitivities to GA, with each cell having a threshold concentration of GA above which it is switched on. The dose-response curve therefore reflects a gradual recruitment of cells to the pool exhibiting a full GA response. Alternatively, all cells may gradually increase their responses as the GA level is increased. In the present study we found that at increasing GA concentrations, increasing numbers of barley (Hordeum vulgare) cells showed the enhanced amylase secretion and vacuolation characteristic of the GA response. We also observed that the region of aleurone tissue closest to the embryo contains the highest proportion of cells activated at the GA concentrations thought to occur naturally in germinating grain. These data indicate that an aleurone layer contains cells of varying sensitivities to GA and that recruitment of these differentially responding pools of cells may explain the broad dose response to GA. PMID- 10364388 TI - Isolation of tobacco isoperoxidases accumulated in cell-suspension culture medium and characterization of activities related to cell wall metabolism. AB - All of the most important guaiacol-type peroxidase (POX) isoforms accumulated in the culture medium of BY-2 tobacco (Nicotiana tabacum L. cv Bright Yellow 2) cells have been isolated. Five basic and two acidic isoforms were found. The four major isoforms (B2, B3, P1, and P2), all strongly basic, have been purified to homogeneity and partially sequenced. B2 and B3 are new isoforms showing high homology to only one POX isolated so far. Amino acid sequencing and specific activities indicated that basic isoPOXs constitute two pairs of strictly related isoforms (P1/P2 and B2/B3). Their specific activities measured in the presence of different substrates, as monolignols and NAD(P)H, indicated possible specialized functions in cell wall metabolism. Only P-type POXs were able to oxidize indoleacetic acid. Variations in pH could play a regulatory role by changing the relative contribution of different isoforms to total POX activity. Apart from cell culture medium, polyclonal antibodies obtained against P1 and P2 detected P1 in roots and in lower parts of stems. Immunocytochemical labeling indicated that P-type POXs were expressed in stem phloem and in phloem and epidermal cells of roots. PMID- 10364389 TI - Molecular and genetic characterization of a novel pleiotropic tomato-ripening mutant AB - In this paper we describe a novel, dominant pleiotropic tomato (Lycopersicon esculentum)-ripening mutation, Cnr (colorless nonripening). This mutant occurred spontaneously in a commercial population. Cnr has a phenotype that is quite distinct from that of the other pleiotropic tomato-ripening mutants and is characterized by fruit that show greatly reduced ethylene production, an inhibition of softening, a yellow skin, and a nonpigmented pericarp. The ripening related biosynthesis of carotenoid pigments was abolished in the pericarp tissue. The pericarp also showed a significant reduction in cell-to-cell adhesion, with cell separation occurring when blocks of tissue were incubated in water alone. The mutant phenotype was not reversed by exposure to exogenous ethylene. Crosses with other mutant lines and the use of a restriction fragment length polymorphism marker demonstrated that Cnr was not allelic with the pleiotropic ripening mutants nor, alc, rin, Nr, Gr, and Nr-2. The gene has been mapped to the top of chromosome 2, also indicating that it is distinct from the other pleiotropic ripening mutants. We undertook the molecular characterization of Cnr by examining the expression of a panel of ripening-related genes in the presence and absence of exogenous ethylene. The pattern of gene expression in Cnr was related to, but differed from, that of several of the other well-characterized mutants. We discuss here the possible relationships among nor, Cnr, and rin in a putative ripening signal cascade. PMID- 10364390 TI - Cold-induced freezing tolerance in Arabidopsis. AB - Changes in the physiology of plant leaves are correlated with enhanced freezing tolerance and include accumulation of compatible solutes, changes in membrane composition and behavior, and altered gene expression. Some of these changes are required for enhanced freezing tolerance, whereas others are merely consequences of low temperature. In this study we demonstrated that a combination of cold and light is required for enhanced freezing tolerance in Arabidopsis leaves, and this combination is associated with the accumulation of soluble sugars and proline. Sugar accumulation was evident within 2 h after a shift to low temperature, which preceded measured changes in freezing tolerance. In contrast, significant freezing tolerance was attained before the accumulation of proline or major changes in the percentage of dry weight were detected. Many mRNAs also rapidly accumulated in response to low temperature. All of the cold-induced mRNAs that we examined accumulated at low temperature even in the absence of light, when there was no enhancement of freezing tolerance. Thus, the accumulation of these mRNAs is insufficient for cold-induced freezing tolerance. PMID- 10364391 TI - Expression of the granule-bound starch synthase I (Waxy) gene from snapdragon is developmentally and circadian clock regulated AB - The granule-bound starch synthase I (GBSSI or waxy) enzyme catalyzes one of the enzymatic steps of starch synthesis. This enzyme is responsible for the synthesis of amylose and is also involved in building the final structure of amylopectin. Little is known about expression of GBSSI genes in tissues other than storage organs, such as seeds, endosperm, and tuber. We have isolated a gene encoding the GBSSI from snapdragon (Antirrhinum majus). This gene is present as a single copy in the snapdragon genome. There is a precise spatial and developmental regulation of its expression in flowers. GBSSI expression was observed in all floral whorls at early developmental stages, but it was restricted to carpel before anthesis. These results give new insights into the role of starch in later reproductive events such as seed filling. In leaves the mRNA level of GBSSI is regulated by an endogenous circadian clock, indicating that the transition from day to night may be accompanied by abolition of expression of starch synthesis genes. This mechanism does not operate in sink tissues such as roots when grown in the dark. PMID- 10364392 TI - Symbiotic root nodules of the actinorhizal plant Datisca glomerata express Rubisco activase mRNA. AB - N2-fixing symbiotic root nodules of the actinorhizal host Datisca glomerata express Dgrca (D. glomerata Rubisco activase) mRNA, a transcript usually associated with photosynthetic organs or tissues. In northern blots a mature, 1700-nucleotide Dgrca mRNA was detected in green plant organs (leaves, flowers, and developing fruits) and in nodules but was not detected in roots. A second message of 3000 nucleotides was observed only in nodules. Both size classes of transcripts were polyadenylated. The larger transcript was 2- to 5-fold more abundant than the mature mRNA; it was hybridized to an intronic probe, indicating that a stable, incompletely spliced transcript was accumulating. Treatment with light on excised nodules did not alter the relative abundance of the two species. In in situ hybridizations the Dgrca message was expressed intensely in the nuclei of infected cells. The Dgrca transcripts also accumulated at lower levels in uninfected cortical cells adjacent to the periderm and the vascular cylinder. mRNA encoding the large subunit of Rubisco (DgrbcL) was abundant in mature infected cells and in the amyloplast-rich sheath of uninfected cortical cells lying between the infected cells and nodule periderm. The proteins Rubisco activase, Rubisco, and the 33-kD O2-evolving complex subunit did not accumulate to detectable levels, indicating that a functional photosynthetic apparatus was not prevalent in nodule tissue. Signals or factors required for the transcription of Dgrca appeared to be present in nodules, but efficient splicing and translation of the message were not observed in Frankia-infected tissue where transcript accumulation was highest. PMID- 10364394 TI - Characterization of photosystem II activity and heterogeneity during the cell cycle of the green alga scenedesmus quadricauda AB - The photosynthetic activity of the green alga Scenedesmus quadricauda was investigated during synchronous growth in light/dark cycles. The rate of O2 evolution increased 2-fold during the first 3 to 4 h of the light period, remained high for the next 3 to 4 h, and then declined during the last half of the light period. During cell division, which occurred at the beginning of the dark period, the ability of the cells to evolve O2 was at a minimum. To determine if photosystem II (PSII) controls the photosynthetic capacity of the cells during the cell cycle we measured PSII activity and heterogeneity. Measurements of electron-transport activity revealed two populations of PSII, active centers that contribute to carbon reduction and inactive centers that do not. Measurements of PSII antenna sizes also revealed two populations, PSIIalpha and PSIIbeta, which differ from one another by their antenna size. During the early light period the photosynthetic capacity of the cells doubled, the O2-evolving capacity of PSII was nearly constant, the proportion of PSIIbeta centers decreased to nearly zero, and the proportion of inactive PSII centers remained constant. During the period of minimum photosynthetic activity 30% of the PSII centers were insensitive to the inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea, which may be related to reorganization of the thylakoid membrane. We conclude from these results that PSII does not limit the photosynthetic activity of the cells during the first half of the light period. However, the decline in photosynthetic activity observed during the last half of the light period can be accounted for by limited PSII activity. PMID- 10364396 TI - Ascorbic acid metabolism in pea seedlings. A comparison of D-glucosone, L sorbosone, and L-galactono-1,4-lactone as ascorbate precursors AB - L-Ascorbic acid (AsA) accumulates in pea (Pisum sativum L.) seedlings during germination, with the most rapid phase of accumulation coinciding with radicle emergence. Monodehydroascorbate reductase and dehydroascorbic acid reductase were active in the embryonic axes before AsA accumulation started, whereas AsA oxidase and AsA peroxidase activities increased in parallel with AsA. Excised embryonic axes were used to investigate the osone pathway of AsA biosynthesis, in which D glucosone and L-sorbosone are the proposed intermediates. [U-14C]Glucosone was incorporated into AsA and inhibited the incorporation of [U-14C]glucose (Glc) into AsA. A higher D-glucosone concentration (5 mM) inhibited AsA accumulation. L Sorbosone did not affect AsA pool size but caused a small inhibition in the incorporation of [U-14C]Glc into AsA. Oxidase and dehydrogenase activities capable of converting Glc or Glc-6-phosphate to glucosone were not detected in embryonic axis extracts. The osones are therefore unlikely to be physiological intermediates of AsA biosynthesis. L-Galactono-1,4-lactone, recently proposed as the AsA precursor (G.L. Wheeler, M.A. Jones, N. Smirnoff [1998] Nature 393: 365 369), was readily converted to AsA by pea embryonic axes. Although L-galactono 1,4-lactone did not inhibit [14C]Glc incorporation into AsA, this does not mean that it is not a precursor, because competition between endogenous and exogenous pools was minimized by its very small pool size and rapid metabolism. PMID- 10364393 TI - Processing, targeting, and antifungal activity of stinging nettle agglutinin in transgenic tobacco. AB - The gene encoding the precursor to stinging nettle (Urtica dioica L. ) isolectin I was introduced into tobacco (Nicotiana tabacum). In transgenic plants this precursor was processed to mature-sized lectin. The mature isolectin is deposited intracellularly, most likely in the vacuoles. A gene construct lacking the C terminal 25 amino acids was also introduced in tobacco to study the role of the C terminus in subcellular trafficking. In tobacco plants that expressed this construct, the mutant precursor was correctly processed and the mature isolectin was targeted to the intercellular space. These results indicate the presence of a C-terminal signal for intracellular retention of stinging nettle lectin and most likely for sorting of the lectin to the vacuoles. In addition, correct processing of this lectin did not depend on vacuolar deposition. Isolectin I purified from tobacco displayed identical biological activities as isolectin I isolated from stinging nettle. In vitro antifungal assays on germinated spores of the fungi Botrytis cinerea, Trichoderma viride, and Colletotrichum lindemuthianum revealed that growth inhibition by stinging nettle isolectin I occurs at a specific phase of fungal growth and is temporal, suggesting that the fungi had an adaptation mechanism. PMID- 10364395 TI - Cloning and characterization of the dihydrolipoamide S-acetyltransferase subunit of the plastid pyruvate dehydrogenase complex (E2) from Arabidopsis. AB - An Arabidopsis cDNA encoding the dihydrolipoamide S-acetyltransferase subunit of the plastid pyruvate dehydrogenase complex (E2) was isolated from a lambdaPRL2 library. The cDNA is 1709 bp in length, with a continuous open reading frame of 1440 bp encoding a protein of 480 amino acids with a calculated molecular mass of 50,079 D. Southern analysis suggests that a single gene encodes plastid E2. The amino acid sequence has characteristic features of an acetyltransferase, namely, distinct lipoyl, subunit-binding, and catalytic domains, although it is unusual in having only a single lipoyl domain. The in vitro synthesized plastid E2 precursor protein has a relative molecular weight of 67,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Upon incubation of the precursor with pea (Pisum sativum) chloroplasts, it was imported and processed to a mature-sized relative molecular weight of 60,000. The imported protein was located in the chloroplast stroma, associated with the endogenous pyruvate dehydrogenase. Catalytically active recombinant plastid E2 was purified as a glutathione S transferase fusion protein. Analysis of plastid E2 mRNA by reverse transcriptase polymerase chain reaction showed highest expression in flowers, followed by leaves, siliques, and roots. The results of immunoblot analysis indicate that protein expression was similar in roots and flowers, less similar in leaves, and even less similar in siliques. This is the first report, to our knowledge, describing a plastid E2. PMID- 10364397 TI - Water content, raffinose, and dehydrins in the induction of desiccation tolerance in immature wheat embryos AB - Desiccation tolerance is initiated in wheat (Triticum aestivum L.) embryos in planta at 22 to 24 d after anthesis, at the time that the embryo water content has decreased from about 73% fresh weight (2.7 g water/g dry weight) to about 65% fresh weight (1.8 g water/g dry weight). To determine if desiccation tolerance is fully induced by the loss of a relatively small amount of water, detached wheat grains were treated to reduce the embryo water content by just a small amount to approximately 69% (2.2 g water/g dry weight). After 24 h of such incipient water loss, subsequently excised embryos were able to withstand severe desiccation, whereas those embryos that had not previously lost water could not. Therefore, a relatively small decrease in water content for only 24 h acts as the signal for the development of desiccation tolerance. Embryos that were induced into tolerance by a 24-h water loss had no detectable raffinose. The oligosaccharide accumulated at later times even in embryos of detached grains that had not become desiccation tolerant, although tolerant embryos (i.e. those that previously had lost some water) contained larger amounts of the carbohydrate. It is concluded that desiccation tolerance and the occurrence of raffinose are not correlated. Immunodetected dehydrins accumulated in embryos in planta as desiccation tolerance developed. Detachment of grains induced the appearance of dehydrins at an earlier age, even in embryos that had not been made desiccation tolerant by incipient drying. It is concluded that a small reduction in water content induces desiccation tolerance by initiating changes in which dehydrins might participate but not by their interaction with raffinose. PMID- 10364398 TI - Transgenic overexpression of the transcription factor alfin1 enhances expression of the endogenous MsPRP2 gene in alfalfa and improves salinity tolerance of the plants AB - Alfin1 cDNA encodes a putative transcription factor associated with NaCl tolerance in alfalfa (Medicago sativa L.). The recombinant protein binds DNA in a sequence-specific manner, including promoter fragments of the NaCl-inducible gene MsPRP2. Alfin1 function was tested in transgenic alfalfa under the control of the 35S promoter in the sense and antisense orientations with the endogenous MsPRP2 as a reporter gene. Calli overexpressing Alfin1 were more resistant to growth inhibition by 171 mM NaCl than vector-transformed controls, whereas calli expressing Alfin1 in the antisense orientation were more sensitive to NaCl inhibition. Transgenic plants overexpressing Alfin1 in the sense orientation grew well. In contrast, the antisense transgenic plants grew poorly in soil, demonstrating that Alfin1 expression is essential for normal plant development. Transgenic calli and plant roots overexpressing Alfin1 showed enhanced levels of endogenous MsPRP2 mRNA accumulation. However, MsPRP2 mRNA accumulation was also regulated in a tissue-specific manner, as shown in leaves of transgenic plants overexpressing Alfin1. These results suggest that Alfin1 acts as a transcriptional regulator in plants and regulates MsPRP2 expression in alfalfa. Alfin1 overexpressing transgenic plants showed salinity tolerance comparable to one of our NaCl-tolerant plants, indicating that Alfin1 also functions in gene regulation in NaCl tolerance. PMID- 10364399 TI - Accumulation of small heat-shock protein homologs in the endoplasmic reticulum of cortical parenchyma cells in mulberry in association with seasonal cold acclimation. AB - Cortical parenchyma cells of mulberry (Morus bombycis Koidz.) trees acquire extremely high freezing tolerance in winter as a result of seasonal cold acclimation. The amount of total proteins in endoplasmic reticulum (ER)-enriched fractions isolated from these cells increased in parallel with the process of cold acclimation. Protein compositions in the ER-enriched fraction also changed seasonally, with a prominent accumulation of 20-kD (WAP20) and 27-kD (WAP27) proteins in winter. The N-terminal amino acid sequence of WAP20 exhibited homology to ER-localized small heat-shock proteins (smHSPs), whereas that of WAP27 did not exhibit homology to any known proteins. Like other smHSPs, WAP20 formed a complex of high molecular mass in native-polyacrylamide gel electrophoresis. Furthermore, not only WAP20 but also 21-kD proteins reacted with antibodies against WAP20. Fractionation of the crude microsomes by isopycnic sucrose-gradient centrifugation revealed that both WAP27 and WAP20 were distributed on a density corresponding to the fractions with higher activity of ER marker enzyme, suggesting localization of these proteins in the ER. When ER enriched fractions were treated with trypsin in the absence of detergent, WAP20 and WAP27 were undigested, suggesting localization of these proteins inside the ER vesicle. The accumulation of a large quantity of smHSPs in the ER in winter as a result of seasonal cold acclimation indicates that these proteins may play a significant role in the acquisition of freezing tolerance in cortical parenchyma cells of mulberry trees. PMID- 10364400 TI - Identification of a cis-regulatory element involved in phytochrome down-regulated expression of the pea small GTPase gene pra2. AB - The pra2 gene encodes a pea (Pisum sativum) small GTPase belonging to the YPT/rab family, and its expression is down-regulated by light, mediated by phytochrome. We have isolated and characterized a genomic clone of this gene and constructed a fusion DNA of its 5'-upstream region in front of the gene for firefly luciferase. Using this construct in a transient assay, we determined a pra2 cis-regulatory region sufficient to direct the light down-regulation of the luciferase reporter gene. Both 5'- and internal deletion analyses revealed that the 93-bp sequence between -734 and -642 from the transcriptional start site was important for phytochrome down-regulation. Gain-of-function analysis showed that this 93-bp region could confer light down-regulation when fused to the cauliflower mosaic virus 35S promoter. Furthermore, linker-scanning analysis showed that a 12-bp sequence within the 93-bp region mediated phytochrome down-regulation. Gel retardation analysis showed the presence of a nuclear factor that was specifically bound to the 12-bp sequence in vitro. These results indicate that this element is a cis-regulatory element involved in phytochrome down-regulated expression. PMID- 10364401 TI - Barley coleoptile peroxidases. Purification, molecular cloning, and induction by pathogens. AB - A cDNA clone encoding the Prx7 peroxidase from barley (Hordeum vulgare L.) predicted a 341-amino acid protein with a molecular weight of 36,515. N- and C terminal putative signal peptides were present, suggesting a vacuolar location of the peroxidase. Immunoblotting and reverse-transcriptase polymerase chain reaction showed that the Prx7 protein and mRNA accumulated abundantly in barley coleoptiles and in leaf epidermis inoculated with powdery mildew fungus (Blumeria graminis). Two isoperoxidases with isoelectric points of 9.3 and 7.3 (P9.3 and P7.3, respectively) were purified to homogeneity from barley coleoptiles. P9.3 and P7.3 had Reinheitszahl values of 3.31 and 2.85 and specific activities (with 2,2'-azino-di-[3-ethyl-benzothiazoline-6-sulfonic acid], pH 5.5, as the substrate) of 11 and 79 units/mg, respectively. N-terminal amino acid sequencing and matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry peptide analysis identified the P9. 3 peroxidase activity as due to Prx7. Tissue and subcellular accumulation of Prx7 was studied using activity-stained isoelectric focusing gels and immunoblotting. The peroxidase activity due to Prx7 accumulated in barley leaves 24 h after inoculation with powdery mildew spores or by wounding of epidermal cells. Prx7 accumulated predominantly in the epidermis, apparently in the vacuole, and appeared to be the only pathogen-induced vacuolar peroxidase expressed in barley tissues. The data presented here suggest that Prx7 is responsible for the biosynthesis of antifungal compounds known as hordatines, which accumulate abundantly in barley coleoptiles. PMID- 10364402 TI - Isolation, chromosomal localization, and differential expression of mitochondrial manganese superoxide dismutase and chloroplastic copper/zinc superoxide dismutase genes in wheat. AB - Superoxide dismutase (SOD) gene expression was investigated to elucidate its role in drought and freezing tolerance in spring and winter wheat (Triticum aestivum). cDNAs encoding chloroplastic Cu/ZnSODs and mitochondrial MnSODs were isolated from wheat. MnSOD and Cu/ZnSOD genes were mapped to the long arms of the homologous group-2 and -7 chromosomes, respectively. Northern blots indicated that MnSOD genes were drought inducible and decreased after rehydration. In contrast, Cu/ZnSOD mRNA was not drought inducible but increased after rehydration. In both spring and winter wheat seedlings exposed to 2 degrees C, MnSOD transcripts attained maximum levels between 7 and 49 d. Transcripts of Cu/ZnSOD mRNA were detected sooner in winter than in spring wheat; however, they disappeared after 21 d of acclimation. Transcripts of both classes of SOD genes increased during natural acclimation in both spring and winter types. Exposure of fully hardened plants to three nonlethal freeze-thaw cycles resulted in Cu/Zn mRNA accumulation; however, MnSOD mRNA levels declined in spring wheat but remained unchanged in winter wheat. The results of the dehydration and freeze thaw-cycle experiments suggest that winter wheat has evolved a more effective stress-repair mechanism than spring wheat. PMID- 10364403 TI - Heterologous expression of a plant small heat-shock protein enhances Escherichia coli viability under heat and cold stress. AB - A small heat-shock protein (sHSP) that shows molecular chaperone activity in vitro was recently purified from mature chestnut (Castanea sativa) cotyledons. This protein, renamed here as CsHSP17. 5, belongs to cytosolic class I, as revealed by cDNA sequencing and immunoelectron microscopy. Recombinant CsHSP17.5 was overexpressed in Escherichia coli to study its possible function under stress conditions. Upon transfer from 37 degrees C to 50 degrees C, a temperature known to cause cell autolysis, those cells that accumulated CsHSP17.5 showed improved viability compared with control cultures. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of cell lysates suggested that such a protective effect in vivo is due to the ability of recombinant sHSP to maintain soluble cytosolic proteins in their native conformation, with little substrate specificity. To test the recent hypothesis that sHSPs may be involved in protection against cold stress, we also studied the viability of recombinant cells at 4 degrees C. Unlike the major heat-induced chaperone, GroEL/ES, the chestnut sHSP significantly enhanced cell survivability at this temperature. CsHSP17.5 thus represents an example of a HSP capable of protecting cells against both thermal extremes. Consistent with these findings, high-level induction of homologous transcripts was observed in vegetative tissues of chestnut plantlets exposed to either type of thermal stress but not salt stress. PMID- 10364405 TI - Phosphoenolpyruvate carboxykinase is involved in the decarboxylation of aspartate in the bundle sheath of maize AB - We recently showed that maize (Zea mays L.) leaves contain appreciable amounts of phosphoenolpyruvate carboxykinase (PEPCK; R.P. Walker, R.M. Acheson, L.I. Tecsi, R.C. Leegood [1997] Aust J Plant Physiol 24: 459-468). In the present study, we investigated the role of PEPCK in C4 photosynthesis in maize. PEPCK activity and protein were enriched in extracts from bundle-sheath (BS) strands compared with whole-leaf extracts. Decarboxylation of [4-14C]aspartate (Asp) by BS strands was dependent on the presence of 2-oxoglutarate and Mn2+, was stimulated by ATP, was inhibited by the PEPCK-specific inhibitor 3-mercaptopicolinic acid, and was independent of illumination. The principal product of Asp metabolism was phosphoenolpyruvate, whereas pyruvate was a minor product. Decarboxylation of [4 14C]malate was stimulated severalfold by Asp and 3-phosphoglycerate, was only slightly reduced in the absence of Mn2+ or in the presence of 3-mercaptopicolinic acid, and was light dependent. Our data show that decarboxylation of Asp and malate in BS cells of maize occurs via two different pathways: Whereas malate is mainly decarboxylated by NADP-malic enzyme, decarboxylation of Asp is dependent on the activity of PEPCK. PMID- 10364404 TI - Enhanced expression and activation of the alternative oxidase during infection of Arabidopsis with Pseudomonas syringae pv tomato. AB - Cyanide-resistant ("alternative") respiration was studied in Arabidopsis during incompatible and compatible infection with Pseudomonas syringae pv tomato DC3000. Total leaf respiration increased as the leaves became necrotic, as did the cyanide-resistant component that was sensitive to salicylhydroxamic acid. Infiltration of leaves with an avirulent strain rapidly induced alternative oxidase (AOX) mRNA, whereas the increase was delayed in the compatible combination. The increase in mRNA correlated with the increase in AOX protein. Increased expression was confined to the infected leaves, in contrast to the pathogenesis-related protein-1, which was induced systemically. Virtually all of the AOX protein was in the reduced (high-activity) form. Using transgenic NahG and mutant npr1-1 and etr1-1 plants, we established that the rapid induction of the AOX was associated with necrosis and that ethylene, but not salicylic acid, was required for its induction. Increased pyruvate levels in the infected leaves suggested that increased substrate levels were respired through the alternative pathway; however, in the control leaves and the infected leaves, respiration was not inhibited by salicylhydroxamic acid alone. Increased respiration appeared to be associated primarily with symptom expression rather than resistance reactions. PMID- 10364406 TI - Influence of white clover mosaic potexvirus infection on the endogenous cytokinin content of bean AB - The cytokinin content in the primary leaves of bean (Phaseolus vulgaris) was monitored for 10 d after inoculation with white clover mosaic potexvirus. The cytokinins were isolated, purified, separated by high-performance liquid chromatography, and quantified by radioimmunoassay. The cytokinins detected at the time of inoculation (d 0) were: (a) the free bases, zeatin (Z), dihydrozeatin (DZ), and isopentenyladenine; (b) the riboside, DZ riboside (DZR); (c) the O glucosides of DZ, DZR, and Z riboside; (d) the nucleotides, Z riboside-5' monophosphate and isopentenyladenosine-5'-monophosphate; and (e) trace amounts of Z-9-glucoside and DZ-9-glucoside. During the 10 d after inoculation with white clover mosaic potexvirus, marked quantitative changes in this cytokinin profile were observed. The concentration of the free bases and DZR decreased, accompanied by an increase in the 9-glucosides and the nucleotides. Virus titer increased rapidly 3 d after inoculation, attaining a maximum level at d 5. This increase coincided with the increases in the 9-glucosides and the nucleotides. We propose that the decline in the cytokinin free bases and riboside may allow the increase of virus titer in bean and lead to the senescence of infected leaves. PMID- 10364407 TI - A gene encoding the cytokinin enzyme zeatin O-xylosyltransferase of Phaseolus vulgaris. AB - Zeatin is the most active and ubiquitous form of the naturally occurring cytokinins. Glycosyl conjugates of zeatin are found in many plant tissues and are considered important for storage and protection against degradative enzymes. Two enzymes catalyzing the formation of O-glycosyl derivatives of zeatin have been characterized, O-glucosyltransferase and O-xylosyltransferase, occurring in seeds of lima bean (Phaseolus lunatus) and bean (Phaseolus vulgaris), respectively. Recently, the ZOG1 gene (zeatin O-glucosyltansferase) was isolated from P. lunatis (). Based on the ZOG1 sequence, the ZOX1 gene (zeatin O xylosyltransferase) was cloned from P. vulgaris. ZOX1 contains an open reading frame of 1362 bp that codes for a 454-amino acid peptide of 51 kD. The recombinant protein has properties identical to the native enzyme: it catalyzes O xylosylzeatin formation with UDP-Xyl as a glycosyl donor but does not recognize UDP-Glucose as a substrate. The ZOX1 and ZOG1 genes exhibit 93% identity at the nucleotide level and 90% similarity at the amino acid level. Neither gene contains introns. These zeatin-specific genes and their promoters will be useful for studies of the regulation of active versus storage forms of cytokinins. Comparison of sequences encoding similar enzymes with distinct substrate specificity may lead to identification of epitopes specific to cytokinin and glycosyl donor molecules. PMID- 10364408 TI - Expression of a gibberellin-induced leucine-rich repeat receptor-like protein kinase in deepwater rice and its interaction with kinase-associated protein phosphatase. AB - We identified in deepwater rice (Oryza sativa L.) a gene encoding a leucine-rich repeat receptor-like transmembrane protein kinase, OsTMK (O. sativa transmembrane kinase). The transcript levels of OsTMK increased in the rice internode in response to gibberellin. Expression of OsTMK was especially high in regions undergoing cell division and elongation. The kinase domain of OsTMK was enzymatically active, autophosphorylating on serine and threonine residues. A cDNA encoding a rice ortholog of a kinase-associated type 2C protein phosphatase (OsKAPP) was cloned. KAPPs are putative downstream components in kinase-mediated signal transduction pathways. The kinase interaction domain of OsKAPP was phosphorylated in vitro by the kinase domain of OsTMK. RNA gel-blot analysis indicated that the expression of OsTMK and OsKAPP was similar in different tissues of the rice plant. In protein-binding assays, OsKAPP interacted with a receptor-like protein kinase, RLK5 of Arabidopsis, but not with the protein kinase domains of the rice and maize receptor-like protein kinases Xa21 and ZmPK1, respectively. PMID- 10364409 TI - Aldehyde oxidase and xanthine dehydrogenase in a flacca tomato mutant with deficient abscisic acid and wilty phenotype AB - The flacca tomato (Lycopersicon esculentum) mutant displays a wilty phenotype as a result of abscisic acid (ABA) deficiency. The Mo cofactor (MoCo)-containing aldehyde oxidases (AO; EC 1.2.3.1) are thought to play a role in the final oxidation step required for ABA biosynthesis. AO and related MoCo-containing enzymes xanthine dehydrogenase (XDH; EC 1.2.1.37) and nitrate reductase (EC 1.6.6.1) were examined in extracts of the flacca tomato genotype and of wild-type (WT) roots and shoots. The levels of MoCo were found to be similar in both genotypes. No significant XDH or AO (MoCo-containing hydroxylases) activities were detected in flacca leaves; however, the mutant exhibited considerable MoCo containing hydroxylase activity in the roots, which contained notable amounts of ABA. Native western blots probed with an antibody to MoCo-containing hydroxylases revealed substantial, albeit reduced, levels of cross-reactive protein in the flacca mutant shoots and roots. The ABA xylem-loading rate was significantly lower than that in the WT, indicating that the flacca is also defective in ABA transport to the shoot. Significantly, in vitro sulfurylation with Na2S reactivated preexisting XDH and AO proteins in extracts from flacca, particularly from the shoots, and superinduced the basal-level activity in the WT extracts. The results indicate that in flacca, MoCo-sulfurylase activity is impaired in a tissue-dependent manner. PMID- 10364410 TI - Purification and characterization of caffeine synthase from tea leaves. AB - Caffeine synthase (CS), the S-adenosylmethionine-dependent N-methyltransferase involved in the last two steps of caffeine biosynthesis, was extracted from young tea (Camellia sinensis) leaves; the CS was purified 520-fold to apparent homogeneity and a final specific activity of 5.7 nkat mg-1 protein by ammonium sulfate fractionation and hydroxyapatite, anion-exchange, adenosine-agarose, and gel-filtration chromatography. The native enzyme was monomeric with an apparent molecular mass of 61 kD as estimated by gel-filtration chromatography and 41 kD as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme displayed a sharp pH optimum of 8.5. The final preparation exhibited 3- and 1-N-methyltransferase activity with a broad substrate specificity, showing high activity toward paraxanthine, 7-methylxanthine, and theobromine and low activity with 3-methylxanthine and 1-methylxanthine. However, the enzyme had no 7 N-methyltransferase activity toward xanthosine and xanthosine 5'-monophosphate. The Km values of CS for paraxanthine, theobromine, 7-methylxanthine, and S adenosylmethionine were 24, 186, 344, and 21 microM, respectively. The possible role and regulation of CS in purine alkaloid biosynthesis in tea leaves are discussed. The 20-amino acid N-terminal sequence for CS showed little homology with other methyltransferases. PMID- 10364411 TI - Carbon uptake and the metabolism and transport of lipids in an arbuscular mycorrhiza AB - Both the plant and the fungus benefit nutritionally in the arbuscular mycorrhizal symbiosis: The host plant enjoys enhanced mineral uptake and the fungus receives fixed carbon. In this exchange the uptake, metabolism, and translocation of carbon by the fungal partner are poorly understood. We therefore analyzed the fate of isotopically labeled substrates in an arbuscular mycorrhiza (in vitro cultures of Ri T-DNA-transformed carrot [Daucus carota] roots colonized by Glomus intraradices) using nuclear magnetic resonance spectroscopy. Labeling patterns observed in lipids and carbohydrates after substrates were supplied to the mycorrhizal roots or the extraradical mycelium indicated that: (a) 13C-labeled glucose and fructose (but not mannitol or succinate) are effectively taken up by the fungus within the root and are metabolized to yield labeled carbohydrates and lipids; (b) the extraradical mycelium does not use exogenous sugars for catabolism, storage, or transfer to the host; (c) the fungus converts sugars taken up in the root compartment into lipids that are then translocated to the extraradical mycelium (there being little or no lipid synthesis in the external mycelium); and (d) hexose in fungal tissue undergoes substantially higher fluxes through an oxidative pentose phosphate pathway than does hexose in the host plant. PMID- 10364412 TI - Protection of photosynthesis against ultraviolet-B radiation by carotenoids in transformants of the cyanobacterium synechococcus PCC7942 AB - The cyanobacterium Synechococcus PCC7942 was transformed with various carotenogenic genes, and the resulting transformants either accumulated higher amounts of beta-carotene and zeaxanthin or showed a shift in the carotenoid pattern toward the formation of zeaxanthin. These transformants were exposed to ultraviolet-B (UV-B) radiation, and the degradation of phycobilins, the inactivation of photosynthetic oxygen evolution, and the activity of photosystem II were determined. In the genetically modified cells, the influence on destruction of phycobilins was negligible. However, protection of photosynthetic reactions against UV-B damage was observed and was dependent on the carotenoid concentrations in the different transformants. Furthermore, it was shown that endogenous zeaxanthin is more effective than beta-carotene. Our results suggest that carotenoids exert their protective function as antioxidants to inactivate UV B-induced radicals in the photosynthetic membrane. PMID- 10364413 TI - Arabidopsis contains at least four independent blue-light-activated signal transduction pathways. AB - We have investigated the stomatal and phototropic responses to blue light of a number of single and double mutants at various loci that encode proteins involved in blue-light responses in Arabidopsis. The stomatal responses of light-grown mutant plants (cry1, cry2, nph1, nph3, nph4, cry1cry2, and nph1cry1) did not differ significantly from those of their wild-type counterparts. Second positive phototropic responses of etiolated mutant seedlings, cry1, cry2, cry1cry2, and npq1-2, were also similar to those of their wild-type counterparts. Although npq1 and single and double cry1cry2 mutants showed somewhat reduced amplitude for first positive phototropism, threshold, peak, and saturation fluence values for first positive phototropic responses of etiolated seedlings did not differ from those of wild-type seedlings. Similar to the cry1cry2 double mutants and to npq1 2, a phyAphyB mutant showed reduced curvature but no change in the position or shape of the fluence-response curve. By contrast, the phototropism mutant nph1-5 failed to show phototropic curvature under any of the irradiation conditions used in the present study. We conclude that the chromoproteins cry1, cry2, nph1, and the blue-light photoreceptor for the stomatal response are genetically separable. Moreover, these photoreceptors appear to activate separate signal transduction pathways. PMID- 10364415 TI - Gibberellin dose-response curves and the characterization of dwarf mutants of barley AB - Dose-response curves relating gibberellin (GA) concentration to the maximal leaf elongation rate (LERmax) defined three classes of recessive dwarf mutants in the barley (Hordeum vulgare L.) 'Himalaya. ' The first class responded to low (10(-8) 10(-6) M) [GA3] (as did the wild type). These grd (GA-responsive dwarf) mutants are likely to be GA-biosynthesis mutants. The second class of mutant, gse (GA sensitivity), differed principally in GA sensitivity, requiring approximately 100 fold higher [GA3] for both leaf elongation and alpha-amylase production by aleurone. This novel class may have impaired recognition between the components that are involved in GA signaling. The third class of mutant showed no effect of GA3 on the LERmax. When further dwarfed by treatment with a GA-biosynthesis inhibitor, mutants in this class did respond to GA3, although the LERmax never exceeded that of the untreated dwarf. These mutants, called elo (elongation), appeared to be defective in the specific processes that are required for elongation rather than in GA signaling. When sln1 (slender1) was introduced into these different genetic backgrounds, sln was epistatic to grd and gse but hypostatic to elo. Because the rapid leaf elongation typical of sln was observed in the grd and gse backgrounds, we inferred that rapid leaf elongation is the default state and suggest that GA action is mediated through the activity of the product of the Sln gene. PMID- 10364414 TI - white anther: A petunia mutant that abolishes pollen flavonol accumulation, induces male sterility, and is complemented by a chalcone synthase transgene AB - A mutation in an inbred line of petunia (Petunia hybrida) produces a reduction in the deep-purple corolla pigmentation and changes the anther color from yellow to white. In addition, the mutant, designated white anther (wha), is functionally male sterile. The inability of pollen from wha plants to germinate in vitro provides a physiological basis for the lack of seed set observed in self-crosses of the mutant. Biochemical complementation with nanomolar amounts of kaempferol, a flavonol aglycone, confirms that the inability of the wha pollen to germinate is due to a lack of this essential compound. Transgenic complementation with a functional ChsA (Chalcone synthase A) cDNA suggests that the genetic lesion responsible for the wha phenotype is in Chs, the gene for the first enzyme in the flavonol biosynthesis pathway. The genetic background of the parental line, as well as the pollen phenotype, allowed us to deduce that the wha mutation is in ChsA. To our knowledge, wha is the first induced, nontransgenic Chs mutant described in petunia, and analysis of the mutation confirms earlier molecular and genetic observations that only two Chs genes (A and J) are expressed in reproductive tissues and that they are differentially regulated in corolla and anther. PMID- 10364416 TI - Muscle substrate utilization and lactate production. AB - Biopsies (biceps) were examined in 8 bodybuilders across a typical arm-curl training session (80% 1-RM). [PCr] and [glycogen] decreased 62 and 12% after 1 set (n = 4), and 50 and 24% after 3 sets (n = 4). [Lactate] was 91 and 118 mmol × kg-1, respectively, after 1 and 3 sets. Fatigue was probably partially caused by decreased [PCr] and increased [H+] (first set) and by decreased [H+] in subsequent sets. PMID- 10364417 TI - Tissue oxygenation by near-infrared spectroscopy and muscle blood flow during isometric contractions of the forearm. AB - The relationship between tissue oxygenation measured by near-infrared spectroscopy (NIRS) and forearm muscle blood flow (FBF) measured by Doppler ultrasound was tested during isometric contractions at 10 and 30% maximal voluntary contraction (MVC) under conditions of normoxia and hypoxia (14% inspired O2). Six subjects maintained contractions at 10% MVC for 5 min and 30% for 2 min in both gas conditions. FBF was elevated during exercise at 10% MVC in hypoxia compared to normoxia, but there was no further increase in flow at 30% MVC. Median power frequency calculations from electromyographic recordings suggested progressive development of fatigue throughout both 10 and 30% MVC contractions. NIRS indicated no change in muscle oxygenation at 10% MVC, but deep venous blood O2 saturation was reduced in normoxia and more so in hypoxia. At 30% MVC, both NIRS and venous O2 saturation were reduced, with no effect of hypoxia on the NIRS signal. While NIRS might provide an indication of muscle oxygenation during isometric exercise, the conflicting findings for NIRS and direct venous blood sampling at 10 vs 30% MVC suggest caution in the application of this noninvasive technique. PMID- 10364418 TI - Etiology of exercise-induced muscle damage. AB - Muscle damage is caused by strenuous and unaccustomed exercise, especially exercise involving eccentric muscle contractions, where muscles lengthen as they exert force. Damage can be observed both directly at the cellular level and indirectly from changes in various indices of muscle function. Several mechanisms have been offered to explain the etiology of the damage/repair process, including mechanical factors such as tension and strain, disturbances in calcium homeostasis, the inflammatory response, and the synthesis of stress proteins (heat shock proteins). Changes in muscle function following eccentric exercise have been observed at the cellular level as an impairment in the amount and action of transport proteins for glucose and lactate/H+, and at the systems level as an increase in muscle stiffness and a prolonged loss in the muscle's ability to generate force. This paper will briefly review factors involved in the damage/repair process and alterations in muscle function following eccentric exercise. PMID- 10364419 TI - Nutritional antioxidants as therapeutic and preventive modalities in exercise induced muscle damage. AB - Several mechanisms have been forwarded to explain the etiology of exercise induced muscle damage. Free-radical mediated processes appear to be an important component of the inflammatory mediated response. Free radicals have also been demonstrated to be a contributing factor in the loss of calcium homeostasis within the cell. Therefore, one of the proposed treatments for preventing or reducing the extent of this damage is the intervention of free-radical mediated processes. Antioxidants are agents that typically work to prevent free-radical mediated alterations within cells by quenching free radicals. The traditional dietary antioxidants most commonly investigated to inhibit free-radical damage are vitamin E, vitamin C, and beta carotene. Other nutritional agents have also been noted to contain antioxidant properties. Isofavonoids and some phytochemicals have been proposed to contain antioxidant properties. This paper briefly reviews some aspects of these agents and the! ! ir role, either proven or proposed, in the prevention of oxidative stress and muscle damage. PMID- 10364421 TI - Chiropractic intervention in the treatment of joint and soft tissue disorders. AB - The concept of manual therapy, specifically manipulation of the bodily joints as in the practice of chiropractic, can no longer be deemed an invalid system of health care. Practiced for over 2,000 years by a variety of ancient civilizations, the art of manipulation for the purpose of correcting and restoring joint function has continued to flourish, despite opposition. The climate, however, is changing. The art of chiropractic is increasingly being seen as a uniquely devised and administered technique whereby high velocity, low amplitude thrusting maneuvers are specifically directed by the skilled practitioner toward spinal segments and peripheral articulations in an effort to correct aberrant mechanical function. The corrections are effected while utilizing the transverse and spinal processes of individual vertebrae as contacting levers. Hippocrates is credited with the advice to, "look well to the spine for the cause of disease," as displaced or degenerative vertebrae m! ! ay irritate spinal nerve roots while exiting the intervertebral foramina and, consequently, interfere with normal nerve function. Similarly, it is a fundamental precept of chiropractic philosophy that irritation of the nervous system by mechanical, chemical, or psychogenic means is considered as causative in the development of disease. The scientific evidence associated with chiropractic intervention in the treatment and management of musculoskeletal disorders and visceral diseases is growing. This paper discusses the history, philosophy, and efficacy of joint manipulation and its influence on the development of chiropractic treatment. PMID- 10364420 TI - Massage and ultrasound as therapeutic modalities in exercise-induced muscle damage. AB - Although both massage and ultrasound treatment are used in clinical settings to enhance muscle functional recovery following exercise-induced muscle damage, there is a paucity of experimental evidence for their efficacy. Theoretically both massage and ultrasound could affect some physiological factors associated with enhancement of postexercise muscle recovery. However, the actual physiological mechanisms by which massage or ultrasound could influence postexercise muscle damage and repair are unknown. Most experimental evidence suggests that massage has little influence on muscle blood flow, clearance of "noxious" substances, recovery of postexercise muscle strength, or delayed soreness sensation. However, more data is needed before conclusions can be drawn as to the ability of massage to influence postexercise inflammatory response or various other physiological changes that characterize exercise-induced muscle damage and repair. There is even less information on the ability of ultrasound to influence physiological or functional factors associated with postexercise muscle damage. The few experiments that have been done tend to be contradictory and have yet to consider the range of ultrasound treatment parameters for therapeutic effectiveness in treating postexercise damage and influencing repair processes. Much more research is needed to determine whether either treatment modality can have any therapeutic effect on exercise-induced muscle damage and recovery of postexercise muscle function. PMID- 10364422 TI - [Evaluation of informed consent in ear, nose, and throat practice]. AB - The patient has a right to information about his or her disease and therapeutic options. A retrospective study was made of the opinion that 40 patients who underwent ear, nose and throat surgery had of this information. After receiving verbal and written information, they completed a questionnaire. Only 1 patient (2.9%) wanted no information, 100% wanted information about the evolution of the disease without treatment, 20.6% were more alarmed after receiving information, and 41.1% thought that written information was essential. PMID- 10364423 TI - Zebrafish in context: uses of a laboratory model in comparative studies. AB - With the recent interest in the reintegration of evolutionary and developmental biology has come a growing need for understanding the phylogenetic relations and degree of generality of the model organisms upon which we rely so heavily. In vertebrate biology the zebrafish Danio rerio has become a paradigmatic system for studies at levels of organization from molecular to interspecific. Studies of model systems in development are often techniques-driven rather than questions based; however, informative hypotheses for developmental research can be derived from phylogenetic distributions of characters. With some understanding of how general the characters of interest are, a thoughtful comparison of the requirements of the questions with the lists of available embryos, reagents, and protocols can guide choices of new vertebrate models. We describe here the phylogenetic placement of zebrafish within the vertebrate world and discuss how generally observations on zebrafish can be taken to apply. We outline a practical protocol for investigating development in a comparative context, illustrated with an example from an ongoing study of teleost tail fin evolution. The principles and procedures presented here apply equally well to any comparative study with an interest in evolution, at any level of phylogeny from intraspecific studies to comparisons across phyla. PMID- 10364424 TI - Notochord-dependent expression of MFH1 and PAX1 cooperates to maintain the proliferation of sclerotome cells during the vertebral column development. AB - During axial skeleton development, the notochord is essential for the induction of the sclerotome and for the subsequent differentiation of cartilage forming the vertebral bodies and intervertebral discs. These functions are mainly mediated by the diffusible signaling molecule Sonic hedgehog. The products of the paired-box containing Pax1 and the mesenchyme forkhead-1 (Mfh1) genes are expressed in the developing sclerotome and are essential for the normal development of the vertebral column. Here, we demonstrate that Mfh1 like Pax1 expression is dependent on Sonic hedgehog signals from the notochord, and Mfh1 and Pax1 act synergistically to generate the vertebral column. In Mfh1/Pax1 double mutants, dorsomedial structures of the vertebrae are missing, resulting in extreme spina bifida accompanied by subcutaneous myelomeningocoele, and the vertebral bodies and intervertebral discs are missing. The morphological defects in Mfh1/Pax1 double mutants strongly correlate with the reduction of the mitotic rate of sclerotome cells. Thus, both the Mfh1 and the Pax1 gene products cooperate to mediate Sonic hedgehog-dependent proliferation of sclerotome cells. PMID- 10364425 TI - Suppression of SHP-2 and ERK signalling promotes self-renewal of mouse embryonic stem cells. AB - The propagation of pluripotent mouse embryonic stem (ES) cells depends on signals transduced through the cytokine receptor subunit gp130. Signalling molecules activated downstream of gp130 in ES cells include STAT3, the protein tyrosine phosphatase SHP-2, and the mitogen-activated protein kinases, ERK1 and ERK2. A chimaeric receptor in which tyrosine 118 in the gp130 cytoplasmic domain was mutated did not engage SHP-2 and failed to activate ERKs. However, this receptor did support ES cell self-renewal. In fact, stem cell colonies formed at 100-fold lower concentrations of cytokine than the unmodified receptor. Moreover, altered ES cell morphology and growth were observed at high cytokine concentrations. These indications of deregulated signalling in the absence of tyrosine 118 were substantiated by sustained activation of STAT3. Confirmation that ERK activation is not required for self-renewal was obtained by propagation of pluripotent ES cells in the presence of the MEK inhibitor PD098059. In fact, the growth of undifferentiated ES cells was enhanced by culture in PD098059. Thus activation of ERKs appears actively to impair self-renewal. These data imply that the self renewal signal from gp130 is a finely tuned balance of positive and negative effectors. PMID- 10364426 TI - Interferon regulatory transcription factors are constitutively expressed and spatially regulated in the mouse lens. AB - Interferon regulatory factors (IRFs) are a family of transcription factors involved in regulation of cell growth and immunological responses. Nine IRFs have been described and they are expressed in a variety of cells, except for ICSBP and LSIRF/Pip, which are thought to be expressed exclusively in immune cells. Here, we show that IRF-1, IRF-2, ICSBP, and LSIRF/Pip are constitutively expressed in the mouse lens. These IRFs are present in both the cytoplasm and the nuclei of lens cells. However, the nuclear and cytoplasmic proteins exhibit distinct mobilities on SDS/PAGE. We further show that in the developing mouse lens, IRF-1 and IRF-2 are expressed at high levels in differentiated lens fiber cells with very low and barely detectable levels in undifferentiated lens epithelial cells. Although the level of ICSBP expression is very low in the normal mouse lens, in transgenic mice with constitutive expression of interferon gamma in the lens, its level is markedly elevated and ICSBP expression is detected exclusively in the nuclei of undifferentiated lens cells. Taken together, our data suggest that expression of IRF transcription factors is spatially regulated in the lens and that distinct IRFs may contribute to differential gene regulation in the epithelial and fiber compartments of the vertebrate lens. PMID- 10364427 TI - Disruption of zebrafish somite development by pharmacologic inhibition of Hsp90. AB - Members of the Hsp90 family of molecular chaperones play important roles in allowing some intracellular signaling molecules and transcription factors to reach and maintain functionally active conformations. In the present study, we have utilized the specific Hsp90-binding agent, geldanamycin, to examine the requirement for Hsp90 during zebrafish development. We show that geldanamycin interacts with both the alpha and the beta-isoforms of zebrafish Hsp90 and that geldanamycin-treated embryos consistently exhibit a number of defects in tissues which express either one of these genes. Within the somites, geldanamycin treatment results in the absence of eng-2-expressing muscle pioneer cells. However, early development of adaxial cells, which give rise to muscle pioneers and which strongly express the hsp90alpha gene shortly before muscle pioneer formation, appeared unaffected. Furthermore, development of the notochord, which provides many of the signals required for proper somite patterning and which does not express detectable levels of either hsp90alpha or hsp90beta mRNA, was similarly unaffected in geldanamycin-treated embryos. The data are consistent with there being a temporal and spatial requirement for Hsp90 function within somitic cells which is necessary for the formation of eng-2-expressing muscle pioneers and possibly other striated muscle fiber types. PMID- 10364428 TI - Altered cell-surface targeting of stem cell factor causes loss of melanocyte precursors in Steel17H mutant mice. AB - The normal products of the murine Steel (Sl) and Dominant white spotting (W) genes are essential for the development of melanocyte precursors, germ cells, and hematopoietic cells. The Sl locus encodes stem cell factor (SCF), which is the ligand of c-kit, a receptor tyrosine kinase encoded by the W locus. One allele of the Sl mutation, Sl17H, exhibits minor hematopoietic defects, sterility only in males, and a complete absence of coat pigmentation. The Sl17H gene encodes SCF protein which exhibits an altered cytoplasmic domain due to a splicing defect. In this paper we analyzed the mechanism by which the pigmentation phenotype in Sl17H mutant mice occurs. We show that in embryos homozygous for Sl17H the number of melanocyte precursors is severely reduced on the lateral neural crest migration pathway by e11.5 and can no longer be detected by e13.5 when they would enter the epidermis in wildtype embryos. The reduced number of dispersing melanocyte precursors correlates with a reduction of SCF immunoreactivity in mutant embryos in all tissues examined. Regardless of the reduced amount, functional SCF is present at the cell surface of fibroblasts transfected with Sl17H mutant SCF cDNA. Since SCF immunoreactivity normally accumulates in basolateral compartments of SCF-expressing embryonic epithelial tissues, we analyzed the localization of wildtype and Sl17H mutant SCF protein in transfected epithelial (MDCK) cells in vitro. As expected, wildtype forms of SCF localize to and are secreted from the basolateral compartment. In contrast, mutant forms of SCF, which either lack a membrane anchor or exhibit the Sl17H altered cytoplasmic tail, localize to and are secreted from the apical compartment of the cultured epithelium. We suggest, therefore, that the loss of melanocyte precursors prior to epidermal invasion, and the loss of germ cells from mature testis, can be explained by the inability of Sl17H mutant SCF to be targeted to the basolateral compartment of polarized epithelial keratinocytes and Sertoli cells, respectively. PMID- 10364429 TI - The timing of lin-4 RNA accumulation controls the timing of postembryonic developmental events in Caenorhabditis elegans. AB - The lin-4 gene encodes a small RNA that is required to translationally repress lin-14 toward the end of the first larval stage of Caenorhabditis elegans development. To determine if the timing of LIN-14 protein down-regulation depends on the temporal profile of lin-4 RNA level, we analyzed the stage-specificity of lin-4 RNA expression during wild-type development and examined the phenotypes of transgenic worms that overexpress lin-4 RNA during the first larval stage. We found that lin-4 RNA first becomes detectable at approximately 12 h of wild-type larval development and rapidly accumulates to nearly maximum levels by 16 h. This profile of lin-4 RNA accumulation corresponded to the timing of LIN-14 protein down-regulation. Transgenic strains that express elevated levels of lin-4 RNA prior to 12 h of development display reduced levels of LIN-14 protein and precocious phenotypes consistent with abnormally early loss of lin-14 activity. These results indicate that the temporal profile of lin-4 RNA accumulation specifies the timing of LIN-14 down-regulation and thereby controls the timing of postembryonic developmental events. PMID- 10364430 TI - Prolactin controls mammary gland development via direct and indirect mechanisms. AB - The inactivation of the prolactin receptor gene by homologous recombination has made it possible to investigate the role of prolactin signaling in mammary gland development without resort to ablative surgery of the endocrine glands. In knockout mice lacking the prolactin receptor, mammary development is normal up to puberty. Subsequently, the ducts branch less frequently than those of wild-type animals. While terminal end buds differentiate to alveolar buds in wild-type females by the end of puberty, in knockout females terminal end bud-like structures persist at the ductal ends. To distinguish between the developmental defects that are intrinsic to the epithelium and those that result from systemic endocrine alterations in prolactin receptor knockout mice, mammary epithelium from prolactin receptor knockouts was transplanted into mammary fat pads of wild type mice. In virgin mice, the knockout epithelial transplants developed normally at puberty, indicating an indirect effect of prolactin on ductal development. Prolactin receptor knockout females are infertile due to multiple reproductive defects, but epithelial transplants allowed us to assess the extent to which the absence of prolactin receptor is limiting, under systemic conditions that allow full mammary gland development. During pregnancy, the prolactin receptor knockout transplants showed normal side branching and the formation of alveolar buds, but no lobuloalveolar development. Thus, prolactin affects mammary morphogenesis in two different ways: it controls ductal side branching and terminal end bud regression in virgin animals via indirect mechanisms, but acts directly on the mammary epithelium to produce lobuloalveolar development during pregnancy. PMID- 10364431 TI - Activin family members in the developing chick retina: expression patterns, protein distribution, and in vitro effects. AB - We have investigated whether the activin family of growth factors is involved in the regulation of retinal cell differentiation. Immunocytochemistry and in situ hybridization have shown that activin/inhibin subunits alpha, betaA, and betaB; receptors II and IIB; follistatin; and a follistatin-like gene are expressed in different regions of the chick embryo retina in developmentally regulated patterns. When tested in dissociated retinal cultures, activin did not appear to affect cell survival or proliferation, but it exerted marked inhibitory effects on the differentiation of photoreceptors, while stimulating the differentiation of nonphotoreceptor neurons; both effects were concentration-dependent and follistatin-sensitive. The results are consistent with the possibility that activin family members play significant roles in the regulation of retinal development. PMID- 10364432 TI - Nuclear entry of the Drosophila melanogaster nuclear lamina protein YA correlates with developmentally regulated changes in its phosphorylation state. AB - The Drosophila melanogaster YA protein is a maternally provided nuclear lamina component that is essential during the transition from meiosis to mitosis at the beginning of embryogenesis. Localization of YA to the nuclear envelope is required for its function; this localization is cell cycle-dependent during embryogenesis. Here we show that the ability of YA to enter nuclei is modulated during development. In developing egg chambers, YA protein is made but excluded from nuclei of nurse cells and oocytes; upon egg activation, YA acquires the ability to enter nuclei and becomes incorporated into the nuclear lamina in unfertilized eggs and embryos. This localization switch correlates with changes in the phosphorylation state of YA. YA in ovaries is hyperphosphorylated relative to YA in unfertilized eggs and embryos. Through site-directed mutagenesis, we identified 443T, a potential phosphorylation site for both cyclin-dependent protein kinase and mitogen-activated-protein kinase, as one of the sites likely involved in this developmental control. Our results suggest that phosphorylation plays a role in modulating the localization of YA during development. A model for developmental regulation of the nuclear entry of YA is proposed and implications for understanding Drosophila egg activation are discussed. PMID- 10364434 TI - SDB 58th annual meeting PMID- 10364433 TI - Developmental expression and cellular origin of the laminin alpha2, alpha4, and alpha5 chains in the intestine. AB - Laminins are extracellular matrix glycoproteins that are involved in various cellular functions, including adhesion, proliferation, and differentiation. In this study, we examine the expression patterns and the cellular origins of the laminin alpha2, alpha4, and alpha5 chains in the developing mouse intestine and in in vitro mouse/chick or chick/mouse interspecies hybrid intestines. In situ hybridization and Northern blot analysis revealed that mRNA levels for all three laminin alpha chains are highest in the fetal intestine undergoing intense morphogenetic movements. Laminin alpha4 mRNA and polypeptide are associated with mesenchyme-derived cell populations such as endothelium and smooth muscle. In contrast, laminin alpha2 and alpha5 chains participate in the structural organization of the subepithelial basement membrane and, in the mature intestine, show a complementary pattern of expression. All three laminin alpha chains occur in the smooth muscle basement membrane, with a differential expression of laminin alpha5 chain in the circular and longitudinal smooth muscle layers. The cellular origin of laminin alpha2 and alpha5 chains found in the subepithelial cell basement membrane was studied by immunocytochemical analysis of mouse/chick or chick/mouse interspecies hybrid intestines at various stages of development using mouse-specific antibodies. Laminin alpha2 was found to be deposited into the basement membrane exclusively by mesenchymal cells, while the laminin alpha5 chain was deposited by both epithelial and mesenchymal cells in an apparently developmentally regulated pattern. We conclude that (1) multiple laminin alpha chains are expressed in the intestine, implying specific roles for individual laminin isoforms during intestinal development, and (2) reciprocal epithelial/mesenchymal interactions are essential for the formation of a structured subepithelial basement membrane. PMID- 10364435 TI - Cellular and molecular bases of axonal regeneration in the fish central nervous system. PMID- 10364436 TI - Neuronal differentiation of NT2/D1 teratocarcinoma cells is accompanied by a loss of lamin A/C expression and an increase in lamin B1 expression. AB - Nuclear lamins are prominent elements of the nuclear matrix and are expressed in cell type-specific and differentiation state-specific patterns. A few observations have indicated that nervous tissue may display unusual patterns of lamin expression, in that some neurons appear to lack A-type lamins, which are generally prominently expressed in terminally differentiated, postmitotic cells. To investigate lamin expression patterns during the differentiation of a teratocarcinoma cell line into neurons, NT2/D1 cells were induced to differentiate with retinoic acid treatment. Lamin expression and organization during differentiation in vitro were examined by quantitative immunofluorescence and immunoblotting methods. Undifferentiated NT2/D1 cells were all strongly labeled with an anti-lamin B1 antibody, but displayed marked variation in A/C lamin immunoreactivity. After differentiation, neuronal nuclear envelopes were significantly more strongly labeled by anti-lamin B1 antibody than those of undifferentiated cells, but completely lacked A/C lamin immunoreactivity. In contrast, nonneuronal cells displayed a slight reduction in B1 lamin immunoreactivity, along with a distinct increase in A/C lamin levels. The loss of lamin A/C expression in NT2/D1 neurons is contrary to the pattern normally observed in most somatic cell types during early development and indicates that the nuclear matrix of some neurons, along with certain neuroendocrine and hematopoietic cells, is uniquely specialized in this regard. PMID- 10364437 TI - Lithium chloride induces the expression of tyrosine hydroxylase in hNT neurons. AB - In the present study, several doses of lithium chloride were tested for their ability to induce the expression of tyrosine hydroxylase (TH) in neurons derived from a human teratocarcinoma cell line (hNT) after 5 and 10 days in vitro (DIV). Following immunocytochemical staining for tyrosine hydroxylase, the percentage of TH-positive neurons was determined and morphometric analysis, including mean soma profile area and neuritic length, was performed. hNT neurons responded to lithium treatment in a dose-dependent manner. In 5 DIV, the most effective dose of lithium chloride (1.0 mM) increased the number of TH-positive neurons approximately sixfold. In addition, both TH-positive hNT neuron mean soma profile area and neurite length were significantly larger than controls by 60 and 70%, respectively. Moreover, even after withdrawal of lithium chloride on day 5, the number of TH-positive neurons in 10 DIV cultures remained significantly increased. These data suggest that hNT cells are indeed responsive to lithium exposure and may serve as a continual source of TH-expressing neurons in new therapeutic approaches to degenerative brain disease. PMID- 10364438 TI - Partial restoration of striatal GABAA receptor balance by functional mesencephalic dopaminergic grafts in mice with hereditary parkinsonism. AB - Levels of inhibitory amino acid receptors were studied in the weaver (wv/wv) mouse model of dopamine (DA) deficiency after unilateral intrastriatal transplantation of fetal mesencephalic cell suspensions. Graft integration was verified by turning behavior tests and from the topographical levels of the DA transporter, tagged autoradiographically with 3 nM [3H]GBR 12935. The average increase in [3H]GBR 12935 binding in grafted dorsal striatum compared to nongrafted wv/wv striatum was 60% 3 months after grafting. Autoradiography of 8 nM [3H]flunitrazepam and 12 nM [3H]muscimol binding was carried out to visualize the distribution of GABAA receptors in +/+ mice and in recipient weaver mutants. A 17% increase in [3H]flunitrazepam binding and a 20% increase in [3H]muscimol binding was found in the nongrafted dorsal striatum of weaver mutants compared to +/+. The functional mesencephalic grafts had a partial normalizing effect on both [3H]flunitrazepam and [3H]muscimol binding in the dorsal striatum of the weaver recipients. The normalization brought about by the grafts was around 20% for [3H]flunitrazepam binding and more than 40% for [3H]muscimol binding. The results are discussed in the context of the important interaction between the converging glutamatergic corticostriatal and DAergic nigrostriatal pathways in controlling the functional GABAergic output of the basal ganglia in Parkinson's disease and in experimental models of DA deficiency. PMID- 10364439 TI - Parabrachial origin of calcitonin gene-related peptide-immunoreactive axons innervating Meynert's basal nucleus. AB - Meynert's basal nucleus is innervated by calcitonin gene-related peptide (CGRP) immunoreactive axons synapsing with cholinergic principal cells. Origin of CGRP immunopositive axons was studied in the albino rat. Since beaded axons containing the nicotinic acetylcholine receptor (nAChR) are also present in the basal nucleus, the microstructural arrangement raises the question whether or not an interaction between CGRP and nAChR exists like in the neuromuscular junction. We found that electrolytic lesion of the parabrachial nucleus results in degeneration of CGRP-immunoreactive axons in the ipsilateral nucleus basalis and induces shrinkage of principal cholinergic neurons while the contralateral nucleus basalis remains intact. Electrolytic lesions in the thalamus, caudate putamen, and hippocampus did not induce alterations in Meynert's basal nucleus. Disappearance of CGRP after lesions of the parabrachial nucleus does not impair presynaptic nAChR in the basal nucleus, suggesting that, unlike in the neuromuscular junction, CGRP is not involved in the maintenance of nAChR in the basal forebrain. It is concluded that the parabrachial nucleus is involved in the activation of the nucleus basalis-prefrontal cortex system, essential in gnostic and mnemonic functions. PMID- 10364440 TI - The presence of isoaspartic acid in beta-amyloid plaques indicates plaque age. AB - Extracellular deposits of fibrillar beta-amyloid are a characteristic neuropathology of Alzheimer's disease (AD). We have developed a novel antibody to a hypothesized "older isomer" of the amyloid protein. This antibody, raised against a synthetic beta-amyloid peptide containing isoaspartic acid at position 7 (isoaspartic-7-Abeta), reacts with isoaspartic-7-Abeta, a nonenzymatic modification found in long-lived proteins. Plaques stained with this antibody are thioflavine positive and are found throughout the frontal and entorhinal cortices of AD cases. In frontal cortex, isoaspartic-7-Abeta plaques are clustered but have a widespread distribution in all cortical layers. Isoaspartic-7-Abeta is found primarily in the core of individual plaques surrounded by nonisomerized amyloid. Activated microglia are associated with plaques containing isomerized and nonisomerized amyloid. In contrast to AD, isoaspartic-7-Abeta plaques in Down's syndrome (DS) cases are found primarily in the superficial layers of frontal cortex. Using image analysis isoaspartic-7-Abeta deposition was correlated with dementia severity in AD and with age in DS. The results indicate that this antibody against altered aspartyl amyloid could be a useful indicator of the age of amyloid plaques. PMID- 10364441 TI - The effect of site and type of nerve injury on spinal glial activation and neuropathic pain behavior. AB - A number of rat peripheral neuropathy models have been developed to simulate human neuropathic pain conditions. The current study sought to determine the relative importance of site versus type of peripheral nerve injury in eliciting mechanical allodynia and spinal glial responses. Rats received one of seven different surgical treatments at the L5 spinal level: spinal nerve cryoneurolysis, spinal nerve tight ligation, dorsal root cryoneurolysis, dorsal root tight ligation, dorsal root transection, ventral root tight ligation, or laminectomy/dural incision sham. Foot-lift response frequency to mechanical stimulation of the ipsilateral hindpaw was assessed postlesion on days 1, 3, 5, and 7. L5 spinal cords were retrieved for immunohistochemical analysis of microglial (OX-42) and astrocytic (anti-glial fibrillary acidic protein) responses. Both types of spinal nerve lesion, freeze and tight ligation, produced rapid and profound mechanical allodynia with intense glial responses. Dorsal root lesions also resulted in intense mechanical allodynia; however, glial responses were almost exclusively astrocytic. Ventral root tight ligation and sham provoked no marked behavioral changes and only sporadic glial responses. Direct dorsal horn communication with the dorsal root ganglion was not a crucial factor in the development of mechanical allodynia, since decentralization of the L5 DRG by complete L5 dorsal root lesion produced profound mechanical sensitization. Conversely, microglial activation responses appear to be dependent upon dorsal root ganglion-mediated signals and, contrary to behavioral responses, were robust only when the lesion was made peripheral to the cell body. Astrocytic activation was always observed following axonal injury and reliably coexisted with behavioral responses. PMID- 10364442 TI - Inducible expression of tryptophan hydroxylase without serotonin synthesis in hypothalamic dopaminergic neurons. AB - In the present study we have further studied the previous findings that rat hypothalamic dopaminergic neuronal cell groups may express tryptophan hydroxylase (TpH), the serotonin synthesizing enzyme, without a detectable serotonin synthesis. Chemical and mechanical neuronal injuries, namely colchicine treatment and axonal transection, respectively, were performed, and distributions of neurons exhibiting immunoreactivity for TpH and/or tyrosine hydroxylase (TH), the dopamine synthesizing enzyme, were analyzed throughout the hypothalamic periventricular and arcuate nuclei. After colchicine treatment there was a statistically significant 87% (P = 0,01) increase in the number of TpH expressing neurons, while TH expression remained essentially similar. Axonal transection resulted also in a statistically significant 131% (P < 0,01) increase in the number of TpH expressing neurons, while TH expression was not significantly altered. All TpH expression coexisted with TH expression, and the induction of TpH expression by neuronal injuries occurred evenly throughout the rostrocaudal length of the territory studied. A possible serotonin synthesis by TpH was examined by giving drugs that increase brain serotonin synthesis, but no immunohistochemically detectable serotonin synthesis could be found in any of the TpH expressing neurons. Finally the possibility was studied that the relative shortage of the cofactor tetrahydrobiopterin would limit serotonin synthesis. However, an administration of tetrahydrobiopterin did not result in detectable serotonin synthesis in these neurons. Taken together these results suggest that dopaminergic neurons in the hypothalamic periventricular and arcuate nuclei are able to express TpH, this expression is induced after neuronal injury, and this induction occurs similarly throughout the territories studied. TpH expression occurs independently of TH expression, and the newly expressed TpH appears not to synthesize serotonin, regardless of pharmacological pretreatments. Thus, our findings (i) support the idea that neurons may possess inducible expression of nonfunctional transmitter-synthesizing enzymes, in this case TpH, and (ii) suggest that expression of an enzyme synthesizing a certain transmitter may not necessarily imply the corresponding transmitter phenotype. PMID- 10364443 TI - Neuronal-glial differential expression of TGF-alpha and its receptor in the dorsal root ganglia in response to sciatic nerve lesion. AB - Injury to peripheral nerves often results in structural and functional changes in the dorsal root ganglia (DRG). Although the mechanisms underlying these changes remain largely unknown, satellite cell activation and up-regulation of several neurotrophic factors in the DRG occur in response to the nerve lesion, modulating the plasticity of affected neurons. To investigate potential roles of transforming growth factor alpha (TGF-alpha) in these plastic changes in the DRG following a sciatic nerve transection, here we examined the expression in DRGs of TGF-alpha and its receptor (EGF receptor), molecules known to be mitogenic to glia and Schwann cells and to be neurotrophic for some differentiated neurons. In the normal DRGs, TGF-alpha and its receptor are expressed mainly in small neurons and satellite cells surrounding some large or medium-sized neurons as determined by immunohistochemistry and in situ hybridization. In response to sciatic nerve lesion, there was a marked and differential up-regulation of TGF-alpha and EGF receptor expression within DRG, evident as early as 24 h after lesion and lasting for at least 14 days. While the up-regulated TGF-alpha was localized mainly on satellite cells in the ipsilateral and contralateral DRGs, EGF receptor up regulation was mainly neuronal (with the expression expanding to include all neurons) in the ipsilateral DRGs, but mainly glial in the contralateral DRGs. These changes in TGF-alpha and its receptor expression suggest that TGF-alpha may play a role in the satellite cell proliferation and/or activation as well as in neuronal survival after nerve lesion. PMID- 10364444 TI - Intracranial injury acutely induces the expression of the secreted isoform of the CNS-specific hyaluronan-binding protein BEHAB/brevican. AB - Hyaluronan (HA) plays an important role in tissue reorganization in response to injury. The mechanisms by which HA participates in these processes are likely to include HA-binding proteins. Previously, we reported the cloning and initial characterization of a central nervous system (CNS)-specific HA-binding protein, BEHAB (brain enriched hyaluronan binding), which was independently cloned in another laboratory and named brevican. BEHAB/brevican mRNA is expressed in the ventricular zone coincident with the initial proliferation and migration of glial cells and in surgical samples of human glioma, where glial-derived cells proliferate and migrate. To determine whether BEHAB/brevican is also expressed during the cellular proliferation and migration associated with CNS injury, we have examined BEHAB/brevican expression during reactive gliosis. BEHAB/brevican occurs as secreted and cell-surface, glycosylphosphatidylinositol (GPI)-anchored, isoforms. The secreted, but not the GPI-anchored, isoform is up-regulated in response to a stab wound to the adult rat brain. The temporal regulation and spatial distribution of BEHAB/brevican expression parallel the gliotic response and the expression of the intermediate filament protein nestin. The up-regulation of BEHAB/brevican in response to CNS injury suggests a role for this extracellular matrix molecule in reactive gliosis. Glial process extension, a central element in the glial response to injury, may require the reexpression of both cytoskeletal and matrix elements that are normally expressed during the glial motility seen in the immature brain. PMID- 10364445 TI - Excitotoxicity plays a role in the death of tyrosine hydroxylase- immunopositive nigral neurons cultured in serum-free medium. AB - We studied the effects of different amino acid receptor antagonists and a calcium (Ca2+) channel blocker on the survival of embryonic tyrosine hydroxylase (TH) immunopositive nigral neurons grown under serum-free culture conditions. Ventral mesencephalic neurons were cultivated for 2 or 7 days. Following serum withdrawal on day 2, some cultures were treated with different concentrations of the noncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist dizocilpine hydrogen maleate (MK-801), the competitive NMDA receptor antagonist 2-amino-5 phosphonopentanoic acid, the competitive kainate/quisqualate receptor antagonist 6,7-dinitroquinoxaline-2, 3-dione, and the Ca2+ channel blocker flunarizine. Treatment with MK-801 or flunarizine increased the survival of TH-positive neurons after serum deprivation. These findings suggest a possible role for excitotoxicity in dopaminergic cell death which can be prevented by blocking the NMDA receptor or by inhibiting Ca2+ entry through voltage-gated channels. PMID- 10364446 TI - Use-dependent exaggeration of brain injury: is glutamate involved? AB - Extreme overreliance on the impaired forelimb following unilateral lesions of the forelimb representation area of the rat sensorimotor cortex (FL-SMC) leads to exaggeration of the initial cortical injury. Glutamate has repeatedly been implicated in the secondary processes leading to neuronal death following traumatic insult, chiefly because of the neuroprotective properties of excitatory amino acid antagonists in a variety of animal models of brain injury. The present study investigated the possibility that NMDA receptor-mediated processes are involved in use-dependent exaggeration of neuronal injury. Rats were fitted with one-sleeved casts that immobilized the intact forelimb for the first 7 days following FL-SMC lesion, a procedure previously shown to result in use-dependent exaggeration of injury and more severe and persistent limb-use deficits. In the present investigation, administration of MK-801 (1 mg/kg ip once daily on alternate days) during the casting period spared neural tissue surrounding the lesion and enhanced functional recovery of the impaired forelimb. These results suggest a role for NMDA receptor-mediated processes in use-dependent exaggeration of injury. PMID- 10364447 TI - alpha-melanocyte-stimulating hormone inhibits NF-kappaB activation and IkappaBalpha degradation in human glioma cells and in experimental brain inflammation. AB - The neuropeptide alpha-melanocyte-stimulating hormone (alpha-MSH) modulates production of proinflammatory cytokines in brain tissue and in peripheral inflammatory cells. Transcription of the genes for these proinflammatory cytokines is regulated by the nuclear factor kappaB (NF-kappaB). NF-kappaB is also activated by proinflammatory cytokines. Degradation of the cytoplasmic inhibitor IkappaBalpha protein results in activation of NF-kappaB. Because of increasing evidence that NF-kappaB is involved in brain injury and inflammation and neurodegenerative disease, we examined whether alpha-MSH inhibits activation of NF-kappaB and limits degradation of IkappaBalpha protein induced by lipopolysaccharide (LPS) in human glioma cells (A-172) and in mouse brain. Electrophoretic mobility shift assays of nuclear extracts from A-172 cells and whole mouse brains stimulated with LPS revealed that alpha-MSH does suppress NF kappaB activation. Western blot analysis demonstrated that alpha-MSH preserved expression of IkappaBalpha protein in vitro (glioma cells) and in vivo (brain tissue). Chloramphenicol acetyltransferase assay indicated that alpha-MSH suppresses NF-kappaB-dependent reporter gene expression induced by LPS in A-172 cells. The findings are consistent with the possibility that the anti inflammatory action of alpha-MSH in CNS inflammation occurs via modulation of NF kappaB activation by peptide-induced inhibition of degradation of IkappaBalpha protein. PMID- 10364448 TI - Pyrithione, a zinc ionophore, inhibits NF-kappaB activation. AB - Pyrrolidine dithiocarbamate (PDTC) suppresses NF-kappaB activity and exhibits cytotoxic effects in bovine cerebral endothelial cells (BCECs), and we have previously reported that these PDTC effects were accompanied by an increase in intracellular zinc levels. To further explore the role of zinc in the modulation of NF-kappaB activation, we studied the effect of pyrithione, a zinc ionophore, on NF-kappaB activation in BCECs. Pyrithione inhibited NF-kappaB activity in a time- and dose-dependent manner. Ca-EDTA, but not Zn-EDTA, prevented pyrithione inhibition of NF-kappaB activity. Pyrithione increased the intracellular zinc level within 15 min. This effect was also abolished by Ca-EDTA, but not by Zn EDTA. The potency of pyrithione on NF-kappaB inhibition and zinc influx was approximately one order of magnitude more potent than PDTC. These findings establish the regulatory role of intracellular zinc levels on NF-kappaB activity in BCECs. PMID- 10364449 TI - DNA binding and transactivating properties of the paired and homeobox protein Pax4. AB - Transcription factors Pax-4 and Pax-6 are known to be key regulators of pancreatic cell differentiation and development. We report on the cloning of a mouse Pax-4 gene, which contains 10 exons, spanning a 4. 7-kbp region. The gene targeting experiments revealed that Pax-4 and Pax-6 cannot substitute for each other in tissue with overlapping expression of both genes. We identified DNA binding specificities of Pax-4 paired domain and paired-type homeodomain. Despite the different Pax-4 amino acid residues in positions responsible for Pax-6 paired domain specificity, the DNA-binding specificities of Pax-4 and Pax-6 are similar. The Pax-4 homeodomain was shown to preferentially dimerize on DNA sequences consisting of an inverted TAAT motif, separated by 4-nucleotide spacing. The Pax 4 transactivation domain was localized within its C-terminal region, which transactivated GAL-based reporter 2.5-fold less than the C-terminal region of Pax 6. We believe that Pax-4 can act as a Pax-6 "repressor," due to the competition for binding sites and lower transactivation potential of Pax-4. PMID- 10364450 TI - Geraniol is a potent inducer of apoptosis-like cell death in the cultured shoot primordia of Matricaria chamomilla. AB - We screened various isoprenoids to search for inducers of apoptosis-like cell death ("apoptosis") in the shoot primordia of Matricaria chamomilla and found that geraniol has the most potent apoptosis-inducing activity among terpenoids, as judged from DNA fragmentation in the cells. The apoptosis-inducing activity of geraniol is concentration and time dependent. Morphologically, the nuclei of the cells treated with 5 mM geraniol were condensed and fragmented. PMID- 10364451 TI - Direct comparison of GAPDH, beta-actin, cyclophilin, and 28S rRNA as internal standards for quantifying RNA levels under hypoxia. AB - The appropriate choice of an internal standard is critical for quantitative RNA analyses. As housekeeping genes, GAPDH, beta-actin, cyclophilin, and 28S rRNA are commonly employed as RNA internal standards with the assumption that their expression levels remain relatively constant in different experimental conditions. We tested this assumption under hypoxia (1% O2, 24 hours) compared to normoxia (20% O2, 24 hours) and compared RNA levels of these 4 housekeeping genes head to head using ribonuclease protection assays. Four biologically diverse cell lines with respect to clonal origin, neoplastic transformation, and growth rates were used in the comparison. Expression levels of 28S rRNA were constant, independent of O2 tension, but levels of GAPDH, beta-actin, and cyclophilin varied widely with hypoxia. In particular, GAPDH mRNA expression was increased by 21.2-75.1% under hypoxic conditions. Increased GAPDH transcription in hypoxia was correlated in the cancer cell lines with upregulation of the transcription factor Hypoxia Inducible Factor-1alpha protein levels in identical experimental conditions. These results suggest that 28S rRNA is a reliable internal control for comparative analyses of transcription under hypoxia; GAPDH appears particularly unfavorable for this purpose either in hypoxia or other experimental conditions that upregulate HIF-1alpha. PMID- 10364452 TI - Detection of exon skipping and retained introns in transcription of the human thrombospondin 2 gene. AB - Alternative transcripts of the gene for human thrombospondin 2 (TSP2), an extracellular glycoprotein, have been identified using a PCR procedure. Species that show exon skipping and/or retained introns were revealed. In particular, a species was characterized in which exon 3 has been spliced from the sequence. This transcript is otherwise intact from exon 1A to exon 22. Transcripts which retain intron 1A, intron 1B, or intron 2 were also identified. Tissue-specific differences were evident in the transcripts with retained introns. In addition, a species which both retained intron 1B and skipped exon 3 was revealed. The existence of these alternative transcripts of the human TSP2 gene could be important for understanding the diverse functions and tissue-specific expression implicated for TSP2. PMID- 10364453 TI - Identification and characterization of a myristylated and palmitylated serine/threonine protein kinase. AB - We report the molecular cloning and initial characterization of a novel fatty acid acylated serine/threonine protein kinase. The putative open reading frame is predicted to encode a 305 amino acid protein possessing a carboxy-terminal protein kinase domain and amino-terminal myristylation and palmitylation sites. The protein kinase has been accordingly denoted as the myristylated and palmitylated serine/threonine protein kinase (MPSK). Human and mouse MPSKs share approximately 93% identity at the amino acid level with complete retention of acylation sites. Radiation hybridization localized the human MPSK gene to chromosome 2q34-37. Northern analysis demonstrated that the human MPSK 1.7 kilobase mRNA is widely distributed. Epitope tagged human MPSK was found to be acylated by myristic acid at glycine residue 2 and by palmitic acid at cysteines 6 and/or 8. Palmitylation of MPSK in these experiments was found to require an intact myristylation site. While epitope tagged MPSK in immune complexes or purified human glutathione S transferase-MPSK was found to autophosphorylate at one or more threonine residues, the enzyme was not found to phosphorylate several other common exogenous substrates. Indeed, only PHAS-I was identified as an exogenous substrate which was found to be phosphorylated on threonine and serine residues. PMID- 10364454 TI - Identification of the major site of in vitro PKA phosphorylation in the polycystin-1 C-terminal cytosolic domain. AB - Sequence analysis of the C-terminal cytosolic domain of human and mouse polycystin-1 has identified three RxS consensus protein kinase A (PKA) phosphorylation motifs. GST-fusion proteins containing the full-length and truncated C-terminal cytosolic domain of murine polycystin-1 were phosphorylated in vitro by the purified catalytic subunit of PKA. This identified a sequence of 25 amino acids, immediately downstream of a previously identified heterotrimeric G-protein activation sequence, as the major site of PKA phosphorylation. Phosphorylation of wild-type and alanine substituted synthetic peptides containing this motif demonstrated that alanine substitution of serine 4159 largely eliminated phosphorylation. Mutation of this residue in the fusion protein reduced phosphorylation by about 70%, whereas mutation of the other two conserved phosphorylation motifs had little effect. We conclude that serine 4159 is the major site of PKA phosphorylation in the C-terminal cytosolic domain of murine polycystin-1. PMID- 10364455 TI - Attenuation of haptoglobin gene expression by TGFbeta requires the MAP kinase pathway. AB - In addition to important roles in the regulation of cell growth and cell restitution, both pro- and anti-inflammatory effects have been ascribed to TGFbeta in intestinal epithelial cells. However, the mechanisms involved in TGFbeta-dependent anti-inflammatory activities remain to be determined. In the rat intestinal epithelial cell line IEC-6, TGFbeta attenuated the glucocorticoid dependent increases in mRNA levels of the acute phase protein gene haptoglobin, and of C/EBP isoforms beta and delta. Supershift assays demonstrated a TGFbeta mediated decrease in the binding of C/EBP isoforms beta and delta to the haptoA and haptoC C/EBP DNA-binding sites from the haptoglobin promoter. Mutations of both HaptoA and HaptoC sites abolished the glucocorticoid-dependent activation and the TGFbeta-mediated attenuation of the haptoglobin promoter, as assessed by transient transfection assays. TGFbeta induced p42/p44 MAP kinase activities. Treatment with the MEK 1/2 inhibitor PD 98059 abolished TGFbeta attenuation. These results suggest that C/EBP isoforms are involved both in the glucocorticoid dependent induction and in the TGFbeta-mediated attenuation of haptoglobin expression. Furthermore, p42/p44 MAP kinases may function in a TGFbeta-dependent signaling pathway leading to attenuation of haptoglobin expression. PMID- 10364456 TI - Regulators of G-protein signaling (RGS) 1 and 16 are induced in response to bacterial lipopolysaccharide and stimulate c-fos promoter expression. AB - Regulators of G-protein signaling (RGS) are negative regulators of G-protein coupled receptor (GPCR) signaling. Sepsis is a pathophysiological condition that is induced primarily in response to bacterial infection and is associated with decreased responsiveness to a number of vasoactive GPCR agonists. Using a degenerate RT-PCR screen, we report that RGS1 and RGS16 were amplified from the heart and aorta of septic animals. By Northern blot analysis, RGS1 and RGS16 were upregulated, with their respective levels increasing 6- and 50-fold in septic hearts. Using a yeast-based bioassay, both RGS1 and RGS16 were found to be equipotent in their ability to attenuate GPCR signaling. These results suggest that both RGS1 and RGS16 contribute to the sepsis-mediated decrease in GPCR signaling. Elevated levels of some RGSs may also lead to an increase in Gbetagamma-activated signaling pathways in the absence of GPCR agonists. Using a c-fos luciferase reporter gene that is responsive to Gbetagamma-activated signaling pathways, we observed a respective 8- and 7-fold increase in the basal luciferase in serum-deprived transfected mammalian cells overexpressing RGS1 or RGS16. This suggests that RGSs play a role in promoting the sepsis-mediated increases in the activation of intracellular signal transduction pathways. PMID- 10364457 TI - Identification of eukaryotic parvulin homologues: a new subfamily of peptidylprolyl cis-trans isomerases. AB - We report here the existence of a subfamily of eukaryotic parvulin proteins that have strong sequence homology with E. coli parvulin, but lack the WW domain found in previously described eukarytoic parvulins. We hence term members of this subfamily EPVH (eukaryotic parvulin homologue). We describe the characterisation of hEPVH (human eukaryotic parvulin homologue). Immunogold labelling transmission electron microscopy reveals that hEPVH is preferentially localised in the mitochondrial matrix. The homology of hEPVH with its prokaryotic ancestor supports the hypothesis that this protein may have a mitochondrial function. An essential role in this organelle may explain the need for a high degree of conservation of this protein between distantly related species. PMID- 10364458 TI - Chromogranin A alters ductal morphogenesis and increases deposition of basement membrane components by mammary epithelial cells in vitro. AB - The extracellular function of chromogranin A (CgA), a glycoprotein widely distributed in secretory vesicles of neurons and neuroendocrine cells, has not been clearly established. To examine whether CgA might modulate the biological properties of epithelial cells, we used an in vitro model of ductal morphogenesis in which mammary epithelial (TAC-2) cells are grown in three-dimensional collagen gels. Whereas under control conditions TAC-2 cells formed thin, branched cords with pointed ends, in the presence of CgA they formed thicker cords with bulbous extremities, reminiscent of growing mammary ducts in vivo. Immunofluorescence analysis demonstrated that CgA increases the deposition of three major basement membrane components, i.e., collagen type IV, laminin, and perlecan, around the surface of the duct-like structures. Similar effects were observed with CgA partially digested with endoproteinase Lys-C, suggesting that one or more fragments of CgA are endowed with the same activity. These findings reveal a hitherto unsuspected activity for CgA, i.e., the ability to alter ductal morphogenesis and to promote basement membrane deposition in mammary epithelial cells. PMID- 10364459 TI - Purification of multisubunit membrane protein complexes: isolation of chloroplast FoF1-ATP synthase, CFo and CF1 by blue native electrophoresis. AB - The proton-ATP synthase of thylakoid membranes from chloroplasts (CFoF1) is composed of two parts with different structural and functional properties: the membrane-integral, proton-conducting complex CFo and the hydrophilic part, CF1 which catalyze the formation of adenosine-5'-triphosphate (ATP). To date it is difficult to isolate functional CFoF1 from thylakoids in high purity and yield. Blue native polyacrylamide gel electrophoresis (BN-PAGE) was therefore successfully employed to isolate CFoF1 in a one-step procedure from thylakoid membranes. Using a cathode buffer with low Coomassie Blue G-250 (CBG) concentration (0.002%), CFoF1 remains intact and can be obtained in high purity from solubilized, prepurified ATP synthase. Using BN-PAGE and a cathode buffer with 0.02% CBG, the ATP synthase bifurcates, and we were able to isolate both parts, CFo and CF1, separately. CFoF1, CFo, and CF1, respectively, were electroeluted nearly quantitatively electroeluted from the gel. BN-PAGE is a generally applicable method for the isolation and characterization of multisubunit membrane protein complexes in their native structure. However, the combination of neutral detergents and the negatively charged dye CBG seems to mimic properties of mild ionic detergents. This effect can lead to dissociation of labile subunits and subcomplexes, especially when delipidated membrane protein complexes are applied to BN-PAGE. By variation of the initial electrophoresis conditions, i.e., dye concentration in the cathode buffer, amount of lipid and detergent, BN-PAGE can be used for the isolation of either intact complexes or of subcomplexes. PMID- 10364460 TI - Functional interaction of poly(ADP-ribose) with the 20S proteasome in vitro. AB - The nuclear enzyme poly(ADP-ribosyl) transferase (pADPRT) catalyzes the formation of poly(ADP-ribose) from NAD+. Several nuclear proteins and pADPRT itself are targets for the modification by poly(ADP-ribosyl)ation. It is demonstrated here that poly(ADP-ribose) or pADPRT automodified with poly(ADP-ribose) interacts noncovalently with the 20S proteasome in vitro. The interaction of pADPRT with the 20S proteasome requires the long ADP-ribose polymers formed by automodification of the pADPRT with poly(ADP-ribose). As a result pADPRT automodified with short ADP-ribose oligomers is unable to associate with the 20S proteasome. The interaction with poly(ADP-ribose) causes a specific stimulation of the peptidase activity of the 20S proteasome. Modified pADPRT does not serve as a substrate for the degradation by the 20S proteasome. No covalent modification of the 20S proteasome by ADP-ribosylation was observed. The results may point to a functional relationship between pADPRT and the 20S proteasome in a pathway protecting the cell from oxidative damage. PMID- 10364461 TI - Smt3, a SUMO-1 homolog, is conjugated to Cdc3, a component of septin rings at the mother-bud neck in budding yeast. AB - SMT3 of Saccharomyces cerevisiae is an essential gene encoding a ubiquitin-like protein similar to mammalian SUMO-1. When a tagged Smt3 or human SUMO-1 was expressed from GAL1 promoter, either gene rescued the lethality of the smt3 disruptant. By indirect-immunofluorescent microscopy, the HA-tagged Smt3 was detected mostly in nuclei and also at the mother-bud neck just like septin fibers. Indeed immunoprecipitation experiments revealed that Cdc3, one of septin components, was modified with Smt3. Furthermore, the protein level of the Cdc3 Smt3 conjugate was reduced and the septin rings disappeared in a ubc9-1 mutant at a restrictive temperature, where the Smt3 conjugation system should be defective. Thus, we conclude that Smt3 was conjugated to Cdc3 in septin rings localized at the mother-bud neck. Around the time of cytokinesis the Cdc3-Smt3 conjugate disappeared. We discuss the biological significance of this Smt3 conjugation to a septin component. PMID- 10364462 TI - Characterization of lectin resistant cell populations derived from human colon carcinoma: correlation of K-Ras with beta1-6 branching of N-linked carbohydrate and CEA production. AB - Previous studies of cell lines derived from human colon carcinoma showed that the extent of beta1-6 branching on N-linked carbohydrate was associated with the presence of K-ras mutation and Ras-activation. We observed that the extent of Ras activation in these cell lines depends not only upon the presence of an activating mutation in K-ras, but also on the amount of total K-Ras protein produced. Here we examined whether negative selective pressure by PHA-L against beta1-6 branching could select for cells having a lower level of K-Ras protein and Ras-activation. PHA-L binds specifically to the beta1-6 branch in N-linked carbohydrate. We utilized a K-ras mutant colon carcinoma cell line, HTB39, which had abundant beta1-6 branching and high levels of K-Ras mutant protein. Lectin resistant cell populations of HTB39 were generated and found to have less beta1-6 branching and less K-Ras protein than their parental counterpart. The lectin resistant cell populations produced lower levels of highly glycosylated CEA, which contributed to the lower level of beta1-6 branching in these cells. PHA-L resistant cell populations were two-fold less sensitive than the parental line to an inhibitor of farnesyl transferase (an enzyme essential for Ras processing and function). This suggested a decrease in dependence on K-ras mediated signaling. Collectively, the data indicated that beta1-6 branching of N-linked carbohydrate and CEA production were linked to K-Ras protein synthesis and activation of the Ras-signaling pathway. PMID- 10364463 TI - Intracellular localization of the Fanconi anemia complementation group A protein. AB - Mutations in the Fanconi anemia (FA) complementation group A (FANCA) gene leads to bone marrow failure, developmental abnormalities and cancer predisposition. To map the intracellular site of FANCA, we constructed a plasmid vector which linked in-frame the enhanced green fluorescent protein (EGFP cDNA) to the 5' end of the FANCA cDNA (pDAS-3). We studied the expression of pDAS-3 in the FANCA mutant fibroblast cell line (GM6914). MMC sensitivity of pDAS-3 transfected cells was comparable to wild-type fibroblasts. The resulting fluorescence pattern in the stable pDAS-3 cell line expressing the fusion protein was primarily nuclear. EGFP selected cells (lacking FANCA) remain hypersensitive to MMC and maintained a cytoplasmic fluorescence pattern. Using deletion mutants of pDAS-3, a nuclear localization domain was identified at the amino terminus of the polypeptide. Western blot results of FANCA protein confirmed the presence of FANCA in nuclear fractions and FANCA protein levels did not vary during cell cycling. This nuclear trafficking of FANCA should guide future work in defining the function of this protein. PMID- 10364464 TI - Selective potentiation of drug cytotoxicity by NSAID in human glioma cells: the role of COX-1 and MRP. AB - Here, we report that nonsteroidal anti-inflammatory drugs (NSAID) enhance the cytotoxic effects of doxorubicin and vincristine in T98G human malignant glioma cells. The cytotoxicity of BCNU, cisplatin, VM26, camptothecin, and cytarabine is unaffected by NSAID. No free radical formation is induced by doxorubicin or vincristine in the absence or presence of NSAID. Doxorubicin and vincristine cytotoxicity in the absence or presence of NSAID are unaffected by free radical scavengers. Functional inhibitors of phospholipase A2 (PLA2), such as dexamethasone and quinacrine, do not mimick the effects of NSAID. T98G cells, but not LN-18, LN-229, LN-308, or A172 glioma cells, express cyclooxygenase (COX-1) and NSAID do not modulate drug cytotoxicity in the other cell lines, except T98G. Thus, augmentation of doxorubicin and vincristine cytotoxicity by NSAID correlates with COX-1 expression. However, ectopic expression of COX-1 in LN-229 cells does not induce the phenotype of T98G cells, indicating that COX-1 inhibition does not mediate the effects of NSAID on drug cytotoxicity. In contrast, a multidrug resistance (MDR) phenotype due to expression of the multidrug resistance-associated protein (MRP) is most prominent in T98G cells and is amenable to modulation by indomethacin, suggesting that inhibition of MRP is at least in partly responsible for the potentiation of doxorubicin and vincristine cytotoxicity by NSAID. PMID- 10364465 TI - Acetate and formate uptake into vesicles isolated from the basolateral region of the plasma membrane of ovine parotid acinar cells. AB - The transport of acetate and formate into plasma membrane vesicles derived from the basolateral face of the ovine parotid acinar cell has an absolute requirement for an anion to be present within the intravesicular space: bicarbonate, formate, acetate, propionate, and butyrate support the uptake of either acetate or formate. A pH gradient across the vesicle membrane, pHi 7.4, pH0 5.5, enhances the uptake of formate, but not acetate. There is no direct relationship between the rate of exchange and the degree of protonation of formate or acetate in the extravesicular medium. The process is saturable and can be inhibited by a range of functional group reagents. When mannitol is the main external osmoticum, the uptake of acetate and formate is still rapid; thus, no other ions are involved in the process apart from the external formate or acetate and the intravesicular anion. This activity could play a major role in the provision of energy in ruminant tissues. PMID- 10364466 TI - CHK down-regulates SCF/KL-activated Lyn kinase activity in Mo7e megakaryocytic cells. AB - The Csk Homologous Kinase (CHK) has been shown to have an enzymatic activity similar to the tyrosine kinase Csk in that it down-regulates Src family kinase activity by causing phosphorylation of the Src C-terminal tyrosine residue. In megakaryocytic Mo7e cells, CHK associates with a specific phosphotyrosine juxtamembrane sequence of the SCF/KL-activated c-Kit receptor. Here, we show that in Mo7e cells, the major Src family kinase activity is p53/56(Lyn). Studies using immobilized c-Kit phosphopeptides show that Lyn is able to specifically associate with the tyrosine-phosphorylated juxtamembrane 568Y*VY*IDPT sequence of c-Kit which has previously been shown to associate with CHK. In cells over-expressing CHK by means of a recombinant vaccinia virus, we observed an elimination of the SCF/KL-stimulated Lyn kinase peak of activity observed at 2-5 minutes in cells infected with the helper T7-expressing vaccinia virus by itself. Examination of total tyrosine phosphorylation by Western blotting showed that over-expression of CHK resulted in a reduction in the levels of tyrosine phosphorylations in the range of 50-60 kDa, but had no apparent effect on c-Kit autophosphorylation. Taken together, these findings show that CHK is able to down-regulate SCF/KL stimulated Lyn activity in megakaryocytes. PMID- 10364467 TI - Activation of telomerase is induced by a natural antigen in allergen-specific memory T lymphocytes in bronchial asthma. AB - The function of the immune system is known to be dependent on the cellular differentiation and clonal expansion of allergen-specific lymphocytes. Telomerase, a ribonucleoprotein enzyme, is believed to be essential for the indefinite proliferation of human cells. To clarify whether telomerase is involved in the pathogenesis of immune diseases as well as of malignancies, we investigated the upregulation of telomerase activity in allergen-specific T lymphocytes. Upregulation of telomerase in allergen-sensitized lymphocytes was induced not only by artificial mitogenic stimulations but also by the natural antigen, house dust mite, which causes allergic diseases. Moreover, the upregulation of telomerase activity in memory T cells activated during allergen specific immune responses might be associated with the enduring allergen-specific atopic propensity in asthmatics. PMID- 10364468 TI - Regulation of leptin by steroid hormones in rat adipose tissue. AB - We investigated if steroid hormones regulate the secretion and the expression of leptin in female and male rat adipose tissue fragments in vitro. Dexamethasone time and dose-dependently increased the secretion and mRNA expression of leptin with a half-maximal stimulation of approximately 1 nM. A time-course revealed a maximal stimulatory effect of 17 beta-estradiol after 24 hours. In male adipose tissue 17 beta-estradiol increased leptin secretion (32% by 50 nM 17 beta estradiol, P = 0.07 and 34% by 500 nM 17 beta-estradiol, P < 1780.05) after 24 hours. An additional effect of estrogen was seen in the dexamethasone (50 nM) stimulated cells (38% with 50 nM 17 beta-estradiol, P < 0.05 and 48% by 500 nM 17 beta-estradiol, P < 0.05). Basal secretion of leptin was equal in female and male adipose tissue, whereas the effects of 17 beta-estradiol (50 nM) and dexamethasone were significantly increased in female as compared with male adipose tissue. Progesterone, testosterone, dihydrotestosterone and dehydroepiandrostendione-sulfate neither affected leptin secretion in male nor female adipose tissue in vitro. Furthermore, to investigate the effect of estrogen female rats were ovariectomized (OVX) and the adipose tissue was incubated in vitro and compared with adipose tissue leptin secretion from sham operated rats (SHAM), and with ovariectomized rats treated with 17 beta-estradiol (EST). A decreased basal and dexamethasone-stimulated leptin secretion from OVX rats compared with SHAM rats was found (P < 0.005) whereas 17 beta-estradiol treatment of ovariectomized rats maintained a normal leptin secretion. However, the dexamethasone stimulation was equally increased above basal levels in SHAM, OVX and EST rats (3.7 +/- 1.2, 2.9 +/- 0.8, 4.2 +/- 1.4, NS, ANOVA) respectively. PMID- 10364470 TI - Effect of steroidal saponins of Anemarrhenae rhizoma on superoxide generation in human neutrophils. AB - Effect of six steroidal saponins isolated from Anemarrhenae rhizoma on superoxide generation in human neutrophils was investigated. The steroidal saponins examined were anemarrhenasaponin-I (An-I), anemarrhenasaponin-Ia (An-Ia), timosaponin B-I (TB-I), timosaponin B-II (TB-II), timosaponin B-III (TB-III) and timosaponin A III (TA-III). An-I, An-Ia, and TB-III suppressed the superoxide generations induced by N-formyl-methionyl-leucyl-phenylalanine (fMLP) and arachidonic acid (AA) in a concentration-dependent manner, but enhanced that induced by phorbol 12 myristate 13-acetate (PMA). While TB-II also suppressed and enhanced the superoxide generations induced by fMLP and PMA, respectively, the compound significantly enhanced the AA-induced superoxide generation. TB-I enhanced the fMLP-induced superoxide generation in a low concentration range (peak at 40 microM), gave no effect on the PMA-induced superoxide generation and weakly enhanced the AA-induced superoxide generation. TA-III enhanced the fMLP-induced superoxide generation more than twice as much as that by TB-I in the same concentration range. However, TA-III enhanced the PMA-induced superoxide generation and most significantly suppressed the AA-induced superoxide generation. PMID- 10364469 TI - A TSH/dibutyryl cAMP activated Cl-/I- channel in FRTL-5 cells. AB - An iodide (I) and chloride (Cl) channel has been identified in the continuously cultured FRTL-5 thyroid cell line using a cell attached patch clamp technique. The channel is activated by TSH and dibutyryladenosine cyclic monophosphate (Bt2 cAMP) but not by phorbol 12-myristate 13-acetate (TPA). Gluconate can not replace chloride or iodide and the channel is impermeable to Na+,K+ and tetraethylammonium ions. The current-voltage relationship demonstrates that the single channel current is a linear function of the clamp voltage. Single channel currents reversed at a pipette potential close to 0 mV. The mean single channel conductance was 60 pS for Cl- and 50 pS for I-. From the I-V relationship there was a strong outward rectification with Cl-, and a complete block with I-, in the single channel current above +40 mV. The feature of the channel is manifested in the single channel records by four distinct, equally spaced conductance levels. We suggest the channel is important for the transport of I and Cl ions across the apical membrane into the colloid space and is important for hormone synthesis and follicle formation. PMID- 10364471 TI - X-Ray structure of glycerol kinase complexed with an ATP analog implies a novel mechanism for the ATP-dependent glycerol phosphorylation by glycerol kinase. AB - Glycerol kinase (GK) catalyzes the Mg-ATP-dependent phosphorylation of glycerol which yields glycerol 3-phosphate. The 2.8 A new crystal structure of GK complexed with an ATP analog revealed an unexpected position of the gamma phosphoryl group, which was 7.2 A distant from the 3-hydroxyl group of glycerol, 5.5 A away from the 3-phosphate of the product (glycerol 3-phosphate) and is stabilized by a beta-hairpin structure. Based on the presented crystal structure and the previously determined structures of GK product complexes, we propose a 3 D model of a nucleophilic in-line transfer mechanism for the ATP-dependent phosphorylation of glycerol by GK. PMID- 10364472 TI - Single integrin molecule adhesion forces in intact cells measured by atomic force microscopy. AB - Cross-talk between cells and the extracellular matrix is critically influenced by the mechanical properties of cell surface receptor-ligand interactions; these interactions are especially well defined and regulated in cells capable of dynamically modifying their matrix environment. In this study, attention was focused on osteoclasts, which are absolutely dependent on integrin extracellular matrix receptors in order to degrade bone; other bone cells, osteoblasts, were used for comparison. Integrin binding forces were measured in intact cells by atomic force microscopy (AFM) for several RGD-containing (Arg-Gly-Asp) ligands and ranged from 32 to 97 picoNewtons (pN); they were found to be cell and amino acid sequence specific, saturatable and sensitive to the pH and divalent cation composition of the cellular culture medium. In contrast to short linear RGD hexapeptides, larger peptides and proteins containing the RGD sequence, such as osteopontin (a major non-collagenous bone protein) and echistatin (a high affinity RGD sequence containing antagonist snake venom protein), showed different binding affinities. This demonstrates that the context of the RGD sequence within a protein has considerable influence upon the final binding force for receptor interaction. These data also demonstrate that AFM, as a methodological approach, can be adapted to cell biology studies wherever cell matrix interactions play a critical role, and, moreover, may have applicability to the analysis of receptor-ligand interactions in cell membranes in general. PMID- 10364473 TI - Matrix-mediated changes in the expression of HNF-4alpha isoforms and in DNA binding activity of ARP-1 in primary cultures of rat hepatocytes. AB - Recently, we have developed a culture system in which rat hepatocytes dedifferentiate and proliferate and after the addition of EHS-gel redifferentiate. During both developmental stages HNF-4alpha2 mRNA was more abundant than HNF-4alpha1 mRNA. However, Western blot analysis using COS-7 cell expressed HNF-4alpha1 and HNF-4alpha2 proteins as standards revealed that (i) HNF 4alpha2 protein was not expressed in dedifferentiated hepatocytes and (ii) either HNF-4alpha2 protein or a highly phosphorylated HNF-4alpha1 protein was the dominating isoform in redifferentiated hepatocytes. The changes in HNF4-isoform expression could not be mimicked by DMSO, suggesting them to be matrix specific. Furthermore, DMSO was less efficient than EHS-gel in reinducing liver-specific gene expression. EHS-gel overlay also led to reduction of ARP-1 DNA binding activity, while overall ARP-1 protein levels did not change. These results suggest that EHS-matrix overlay regulates the expression of different HNF-4alpha isoforms on a posttranscriptional level while ARP-1 DNA binding activity is regulated by posttranslational mechanisms. PMID- 10364474 TI - Millisecond analyses of Ca2+ initiation sites evoked by muscarinic receptor stimulation in exocrine acinar cells. AB - High speed laser confocal microscopy (8 ms/image) was applied to the dissociated parotid acini as a model to study Ca2+ signaling mechanisms in non-excitable exocrine secretory cells. Immunofluorescence microscopy showed the localization of IP3 receptor type 2 along the apical membrane region. Muscarinic stimulation with carbachol evoked a rise in [Ca2+]i that was initiated from apical region and propagated into basal region as Ca2+ waves. This was most clearly observed when extracellular Ca2+ was omitted. Carbachol also triggered the abrupt increase of [Ca2+]i simultaneously at both basal and apical regions in many acini. Within an acinus, each cell responded synchronously. The present results suggest that one Ca2+ initiation site in the rat parotid acinar cell is apical region, corresponding to the localization of IP3 receptors. Another Ca2+ initiation site is basal region, which seems to be related to Ca2+ entry from extracellular medium and/or Ca2+ release from basally located organelles such as nuclei and endoplasmic reticulum. PMID- 10364475 TI - Store Ca2+ depletion enhances NMDA responses in cultured human astrocytes. AB - NMDA produced whole-cell membrane currents in cultured human astrocytes. The currents were not inhibited by the selective NMDA receptor antagonist, APV, while they were partially inhibited by the broad G-protein inhibitor, GDPbetaS. NMDA induced currents were enhanced by either the microsomal Ca2+/ATPase inhibitors, thapsigargin and cyclopiazonic acid, or the ATP-uncoupler, dinitrophenol (DNP). In the Ca2+ assay, NMDA increased intracellular calcium concentration. The increase was inhibited by 26% in Ca2+-free extracellular solution, and it was not inhibited by APV. The results of the present study suggest that NMDA responses in human astrocytes are regulated by store Ca2+ depletion-associated signal. PMID- 10364477 TI - The endothelial cell protein C receptor (EPCR) functions as a primary receptor for protein C activation on endothelial cells in arteries, veins, and capillaries. AB - Plasma protein C functions as an anticoagulant when it is converted to the active form of serine protease. Protein C activation has been found to be mediated by the endothelial cell surface thrombin/thrombomodulin (TM) complex. In addition, we recently identified the endothelial cell protein C/activated protein C receptor (EPCR) which is capable of high-affinity binding for protein C. In this study, we established monoclonal antibodies (mAbs) against EPCR including several function blocking antibodies. Immunohistochemical analysis using these mAbs demonstrated that EPCR is widely expressed in the endothelial cells of arteries, veins, and capillaries in the lung, heart, and skin. Function blocking anti-EPCR mAbs strongly inhibited protein C activation mediated by primary cultured arterial endothelial cells which express abundant EPCR. Anti-EPCR mAbs also prevent protein C activation mediated by microvascular endothelial cells. These results indicate that EPCR functions as an important regulator for the protein C pathway in various types of vessels. PMID- 10364476 TI - Characterization of DNA binding properties and sequence specificity of the human 52 kDa Ro/SS-A (Ro52) zinc finger protein. AB - The Ro52 protein is an autoantigen in Sjogren's Syndrome and systemic lupus erythematosus. Although its function is not yet known, sequence similarities to other proteins suggest that it binds to DNA. In this study, the hypothesis that Ro52 recognizes particular nucleic acid sequences was tested. Ro52 bound to double stranded but not single stranded DNA. 1,10-Phenanthroline, a chelater of zinc, was found to inhibit this interaction, suggesting that the zinc fingers of Ro52 are functional. DNA sequences were selected from an oligonucleotide library by binding to Ro52 followed by amplification by Taq DNA polymerase in order to characterize the DNA sequence-binding motif for this protein. These studies support the hypothesis that Ro52 is functionally a member of a family of zinc finger proteins, many of which are known to bind to DNA or regulate gene expression. We speculate that Ro52 functions as a transcription factor, and that its disregulation may have important consequences in the expression or susceptibility of certain autoimmune diseases. PMID- 10364478 TI - 3,4-methylenedioxymethamphetamine ("Ecstasy") stimulates the expression of alpha1(I) procollagen mRNA in hepatic stellate cells. AB - 3,4-Methylenedioxymethamphetamine, MDMA ("Ecstasy"), has been previously shown to produce cell necrosis and fibrosis in the liver. Our aim was to study the effect of MDMA on the type I collagen production by a cell line of hepatic stellate cells (HSC), the cell type mainly responsible for collagen synthesis in the liver. We demonstrated that MDMA increases alpha1(I) procollagen mRNA levels and that this increase correlates with glutathione depletion and enhanced hydrogen peroxide production by HSC. Pre-treatment with either glutathione monoethyl ester or deferoxamine prevents the MDMA-induced alpha1(I) procollagen mRNA expression, indicating oxidative stress to be a mediator of this effect. Lipid peroxidation was not detected in MDMA-treated cells and therefore does not seem to be involved in the pro-fibrogenic action of MDMA on HSC. PMID- 10364479 TI - Effects of exogenous human heparin-binding epidermal growth factor-like growth factor on DNA synthesis of hepatocytes in normal mouse liver. AB - Heparin-binding epidermal growth factor-like growth factor (HB-EGF) has been reported to stimulate DNA synthesis of the hepatocytes in culture and highly express in regenerating rat liver after partial hepatectomy. We examined mitogenic effects and activation of transcription factors caused by exogenous human HB-EGF (hHB-EGF) in mouse liver. The mean labeling index in hepatocytes of hHB-EGF-injected mice was 2.6%, a significant increase over that in saline injected controls (under 0.01%). By exogenous hHB-EGF injection, activation of transcription factors such as nuclear factor (NF)-kappaB and activator factor (AP)-1 was observed in the liver. By Northern blot analysis, hepatocyte growth factor (HGF) gene expression in the liver was found to be induced in the hHB-EGF injected mice. In conclusion, intravenously injected hHB-EGF showed a limited but definite effect on the DNA synthesis of hepatocytes in the mice liver. HB-EGF may serve as a hepatotrophic factor in vivo. PMID- 10364480 TI - HMG CoA reductase and LDL receptor genes are regulated differently by 15 ketosterols in Hep G2 cells. AB - The ability of two 15-ketosubstituted sterols, 5alpha-cholest-8(14)-en-3beta-ol 15-one and 3beta-(2-hydroxyethoxy)-5alpha-cholest-8(14)-en-15-one, to alter the mRNA levels of 3-hydroxy-3-methylglutaryl-CoA reductase, low density lipoprotein receptor, and oxysterol binding protein was studied and compared with the effects of 25-hydroxycholesterol in Hep G2 cells. All three oxysterols decreased the level of HMG CoA reductase mRNA at concentrations of 10-30 microM, although 25 hydroxycholesterol was effective at concentrations of 1-3 microM. 25 Hydroxycholesterol lowered the level of LDL receptor mRNA more efficiently after 8 hours than after 24 hours of incubation, whereas 15-ketosterols did not decrease the mRNA level of the LDL receptor. The transcriptions of HMG CoA reductase and LDL receptor genes are therefore independently regulated by 15 ketosterols in Hep G2 cells. In addition, the level of the oxysterol binding protein mRNA is not affected by oxysterols in Hep G2 cells. PMID- 10364481 TI - Presence of leptin in breast cell lines and breast tumors. AB - Leptin is the product of the ob gene, reported to be secreted exclusively from adipocytes and thought to control satiety by providing information to the central nervous system. However, the function of leptin appears to be more complex because multiple studies demonstrate its role in hematopoiesis, reproduction, and immunity. In addition, several nonadipose sources of leptin have been reported. The purpose of this study was to examine several breast cancer cell lines and ductal carcinomas of the breast for expression of leptin messenger RNA (mRNA) and protein. For tumor studies, specimens were preassayed for contaminating adipose tissue. Northern blot analyses demonstrated leptin mRNA in several breast cancer cell lines (MCF-7, T47D, and MDA-MB-231), a normal breast epithelial cell line (MCF10A), and four breast tumors. Leptin protein was identified in T47D breast cancer cells by indirect immunofluorescent staining and in samples of the same breast tumors used for Northern studies by enzyme-linked immunosorbent assays (ELISA). This preliminary study suggests that leptin is expressed in malignant epithelial cells of the breast. Further investigation is needed to determine whether this protein plays a role in breast carcinogenesis. PMID- 10364482 TI - Male sterility in transgenic mice expressing activin betaA subunit gene in testis. AB - Activins and inhibins, which are endocrine regulators of anterior pituitary function, have also been reported to participate in the paracrine and autocrine regulation of reproductive function. To determine the in vivo effects of overexpressed activin/inhibin, we generated transgenic mice carrying the human activin/inhibin betaA subunit mini gene under the regulatory control of the mouse methallothionein promoter. In one of the transgenic line analyzed, the betaA subunit gene was preferentially expressed in the testis. Ectopic and allochronic expression of the betaA gene started at 3 weeks after birth and transgenic male mice became sterile in the ensuing several weeks. Histological analysis revealed testicular degeneration in these mice. The results from this transgenic line strongly support the in vivo activity of activin/inhibin in male reproductive functions. PMID- 10364483 TI - Lens proteins block the copper-mediated formation of reactive oxygen species during glycation reactions in vitro. AB - The formation of advanced glycation endproducts (AGEs) from glucose in vitro requires both oxygen and a transition metal ion, usually copper. These elements combine to produce reactive oxygen species (ROS) which degrade glucose to AGE forming compounds. We measured the ability of Cu(2+) to accelerate ROS formation, and the effect of added lens proteins on these reactions. Increasing levels of Cu(2+) accelerated the formation of superoxide anion with glucose and fructosyl lysine, but the addition of 2.0 mg/ml calf lens proteins completely blocked superoxide formation up to 100 microM of added Cu(2+). Lens proteins, however, had no effect on superoxide generated by the hypoxanthine/xanthine oxidase system. The oxidation of ascorbic acid was increased 170-fold by the addition of 10 microM Cu(2+), but was also completely prevented by added lens proteins. Hydroxyl radical formation, as measured by the conversion of benzoate to salicylate, was increased to 30 nmoles/ml after 18 h by the addition of 100 microM Cu(2+) and 2.5 mM H2O2. This increase was also blocked by the addition of lens proteins. However, hydroxyl radical formation, as estimated by the crosslinking and fragmentation of lens proteins, was observed in the presence of 100 microM Cu(2+), likely at the sites of Cu(2+) binding. Since the ratio of lens proteins to Cu(2+) in human lens is at least 1000-fold higher than those used here, the data argue that Cu(2+) in the lens would be tightly bound to protein, preventing ROS-mediated AGE formation from glucose in vivo. PMID- 10364484 TI - A mechanism of resistance to HIV-1 entry: inefficient interactions of CXCR4 with CD4 and gp120 in macrophages. AB - To test the hypothesis that inefficient interactions of CXCR4 with CD4 and gp120 could affect HIV-1 entry, we incubated macrophages, monocytes, and lymphocytes with gp120 and coimmunoprecipitated CD4 by using anti-CXCR4 antibodies. CD4 was efficiently coimmunoprecipitated in lymphocytes and monocytes but not in macrophages. Overexpression of CD4 in macrophages resulted in detection of CD4 CXCR4 and gp120-CD4-CXCR4 complexes in parallel with the restoration of macrophage fusion susceptibility. These results suggest a mechanism of resistance to entry of some X4 HIV-1 strains into macrophages and a method for dissection of the initial stages of HIV entry. PMID- 10364485 TI - Platelets from thrombocytopenic ponies acutely infected with equine infectious anemia virus are activated in vivo and hypofunctional. AB - Thrombocytopenia is a consistent finding and one of the earliest hematological abnormalities in horses acutely infected with equine infectious anemia virus (EIAV), a lentivirus closely related to human immunodeficiency virus. Multifactorial mechanisms, including immune-mediated platelet destruction and impaired platelet production, are implicated in the pathogenesis of EIAV associated thrombocytopenia. This study was undertaken to investigate whether regenerative thrombopoiesis and platelet destruction occurred in ponies acutely infected with EIAV. Circulating large, immature platelets were increased in ponies acutely infected with EIAV late in the infection when platelet count was at a nadir. Morphometric analysis of bone marrow from acutely infected ponies revealed significant increased in megakaryocyte area and megakaryocyte nuclear area. A trend toward increased numbers of megakaryocytes was also observed. Platelets from acutely infected ponies had increased surface-bound fibrinogen and ultrastructural changes consistent with in vivo platelet activation. Platelets also had hypofunctional aggregation responses to three agonists in vitro. We conclude that thrombocytopenia in ponies acutely infected with EIAV is regenerative and suggest that bone marrow platelet production is not severely compromised in these ponies. Our findings reveal that in vivo platelet activation occurs in ponies acutely infected with EIAV, and as a result platelets are hypofunctional in vitro. Activation of platelets in vivo may cause platelet degranulation or formation of platelet aggregates, which would result in removal of these damages platelets from circulation. This may represent a form of nonimmune-mediated platelet destruction in ponies acutely infected with EIAV. PMID- 10364486 TI - The transcriptional activation domain of VP16 is required for efficient infection and establishment of latency by HSV-1 in the murine peripheral and central nervous systems. AB - The herpes simplex virus (HSV) transactivator VP16 is a structural component of the virion that activates immediate-early viral gene expression. The HSV-1 mutant in1814, which contains a 12-bp insertion that compromises the transcriptional function of VP16, replicated to a low level if at all in the trigeminal ganglia of mice (I. Steiner, J. G. Spivack, S. L. Deshmane, C. I. Ace, C. M. Preston, and N. W. Fraser (1990). J. Virol. 64, 1630-1638; Valyi-Nagy et al., unpublished data). However, in1814 did establish a latent infection in the ganglia after corneal inoculation from which it could be reactivated. In this study, several HSV-1 strains were constructed with deletions in the VP16 transcriptional activation domain. These viruses were viable in cell culture, although some were significantly reduced in their ability to initiate infection. A deletion mutant completely lacking the activation domain of VP16 (RP5) was unable to replicate to any detectable level or to efficiently establish latent infections in the peripheral and central nervous systems of immunocompetent mice. However, similar to in1814, RP5 formed a slowly progressing persistent infection in immunocompromised nude mice. Thus RP5 is severely neuroattenuated in the murine model of HSV infection. However, the activation domain of VP16 is not essential for replication in the nervous system, since we observed a slow progressive infection persisting in the absence of an immune response. PMID- 10364487 TI - Mutational analyses of the putative calcium binding site and hinge of the turnip crinkle virus coat protein. AB - The turnip crinkle carmovirus (TCV) coat protein (CP) is folded into R (RNA binding), S (shell), and P (protruding) domains. The S domain is an eight stranded beta barrel common to the coat protein subunits of most RNA viruses. A five-amino-acid hinge connects the S and P domains. In assembled particles, each pair of CP subunits is thought to bind a single calcium ion through interactions with three residues of one subunit and two residues of a neighboring subunit. These five residues comprise the putative calcium-binding site (CBS). The putative CBS and hinge are adjacent to one another. Mutations were introduced into the putative CBS or hinge in an effort to further determine the biological functions of TCV CP. One putative CBS mutant, TCV-M32, exhibited wild-type cell to-cell movement but failed to move systemically in Nicotiana benthamiana, and particles were not detected. Another putative CBS mutant, TCV-M23, exhibited deficient cell-to-cell movement but particles accumulated in isolated protoplasts. Two other putative CBS mutants, TCV-M22 and -M33, showed wild-type cell-to-cell and systemic movement but elicited mild systemic symptoms that were somewhat delayed. All of the hinge mutants exhibited wild-type movement but some elicited non-wild-type symptoms. Point mutations in the putative CBS or hinge appear to alter virus-ion interactions, secondary structure, or particle conformation, thereby affecting interactions between the CP and plant hosts. PMID- 10364488 TI - Sequence requirements for the nuclear localization of the murine cytomegalovirus M44 gene product pp50. AB - The murine cytomegalovirus (MCMV) M44 gene product pp50 is normally present in the nuclei of virus-infected cells. During transient expression of pp50 in COS-1 cells, the phosphoprotein was readily detectable in the nuclei, indicating that it possesses a nuclear localization signal (NLS). Studies on the subcellular locations of N- and C-terminal deletion mutants of pp50 suggested that alterations in both the C terminus and the highly conserved N-terminal domains of pp50 affect nuclear localization. In particular, the C-terminal 11 amino acids of pp50, which includes a "KKQK" motif, were able to mediate the import of a beta galactosidase fusion protein into the nucleus. The pair of lysine residues in this motif constitutes an essential element of the C-terminal NLS as mutation of this motif to AAQK directly affected the nuclear localization of either pp50 or beta-galactosidase fusion proteins containing the C-terminal portion of pp50. Furthermore our results indicated that the functionality of the C-terminal NLS is dependent on the structural integrity of the highly conserved N-terminal portion of the molecule, as deletion of amino acids 157-201 alone adversely affected nuclear localization. In the absence of a functional C-terminal NLS, the subcellular localization of pp50 is sensitive to potential conformational changes induced by mutations within the N-terminal half of the molecule. Under those circumstances, mutation of the YK residues at position 22-23 or deletion of amino acids 267-283 was sufficient to produce a protein that was impaired in nuclear import or retention. PMID- 10364489 TI - Biochemical and functional properties of a palindromic sequence motif within the hepatitis B virus enhancer 1. AB - The hepatitis B virus (HBV) enhancer 1 is a transcriptional element that contributes to the liver-specific regulation of HBV gene expression. We previously identified a novel protein binding site within the enhancer that contains an 8-bp palindromic sequence motif. This motif partially overlaps the binding sites for nuclear factor 1 and hepatocyte nuclear factor 3beta (HNF3beta). Moreover, we demonstrated that this novel site is recognized by a protein or proteins, tentatively designated as palindrome-binding factor (PBF), that cooperatively interact with HNF3beta. In the present work, we have further examined the biochemical and functional attributes of PBF. Protein-DNA interaction studies indicate that three thymidine residues located at the 3'-end of the palindromic sequence motif are important for maximal PBF-binding activity. When protein-DNA complexes were photocrosslinked by exposure to ultraviolet (UV) light, a prominent polypeptide with an apparent molecular mass of 50 kDa was found to associate with the PBF-binding site. Furthermore, transient transfection studies support the hypothesis that PBF contributes to enhancer 1 activity by a combinatorial mechanism that involves at least one other cis-acting sequence motif, the HNF3beta-binding site. PMID- 10364491 TI - Involvement of a p53-dependent pathway in rubella virus-induced apoptosis. AB - In light of the important role of apoptotic cell death in the pathogenesis of several viral infections, we asked whether the cytopathogenicity evoked by rubella virus (RV) might also involve apoptotic mechanisms. The To-336 strain of RV induced apoptosis in Vero and RK-13 cells, but not in fibroblast cell lines. UV-inactivated RV virions did not elicit the apoptotic response, indicating that productive infection is required for the induction of cell death. Both p53 and p21 protein levels were highly elevated in RV-infected Vero cells. The level of p21 mRNA was increased, while expression of the p53 gene was unaffected by RV infection. A dominant-negative p53 mutant (p53(W248)) conferred partial protection from RV-induced apoptosis. These data implicate a p53-dependent apoptotic pathway in the cytopathogenicity of RV, thereby suggesting a mechanism by which RV exerts its teratogenic effects. PMID- 10364490 TI - Goat milk epithelial cells are highly permissive to CAEV infection in vitro. AB - The main route of small ruminant lentivirus dissemination is the ingestion of infected cells present in colostrum and milk from infected animals. However, whether only macrophages or other cell subtypes are involved in this transmission is unknown. We derived epithelial cell cultures, 100% cytokeratin positive, from milk of naturally infected and noninfected goats. One such culture, derived from a naturally infected goat, constitutively produced a high titer of virus in the absence of any cytopathic effect. The other cultures, negative for natural lentivirus infection, were tested for their susceptibility to infection with the CAEV-CO strain and a French field isolate CAEV-3112. We showed that milk epithelial cells are easily infected by either virus and produce viruses at titers as high as those obtained in permissive goat synovial membrane cells. The CAEV-CO strain replicated in milk epithelial cells in absence of any cytopathic effect, whereas the CAEV-3112 field isolate induced both cell fusion and cell lysis. Our results suggest that CAEV-infected milk epithelial cells of small ruminants may play an important role in virus transmission and pathogenesis. PMID- 10364492 TI - Bovine viral diarrhea virus quasispecies during persistent infection. AB - Analysis of viral genome sequences from two calves persistently infected with bovine viral diarrhea virus revealed a quasispecies distribution. The sequences encoding the glycoprotein E2 were variable, translating to a number of changes in predicted amino acid sequences. The NS3 region was found to be highly conserved in both animals. The number of E2 clones showing variant amino acids increased with the age of the animal and comparison of the consensus sequences at the different time points confirmed differences in the predicted E2 sequences over time. The immune tolerance that allows the lifelong persistence of this viral infection is highly specific. It is likely that some of the variant viruses generated within these animals will differ antigenically from the persisting virus and be recognized by the immune system. Evidence of an immune response to persisting virus infection was gathered from a larger sample of cattle. Serum neutralizing antibodies were found in 4 of 21 persistently infected animals. Accumulations of viral RNA in the lymph nodes of all animals examined, particularly in the germinal center light zone, may represent antigenic variants held in the form of immune complexes on the processes of follicular dendritic cells. PMID- 10364493 TI - High diversity of HIV-1 subtype F strains in Central Africa. AB - In this paper, we studied the variability of HIV-1 subtype F strains in Africa. For 11 viruses, mainly of Central African origin, different parts of the genome were genetically characterized. For all strains the V3-V5 region of the envelope gene was sequenced, and for 7 strains, the entire envelope gene was studied. For 10 strains, the p24 region of the gag gene was also sequenced. For each region studied, three subgroups in the F subtype were identified, F1, F2, and F3. These three subgroups were supported by high bootstrap values and the intra- and inter subgroup F distances were comparable to those obtained for the known subtypes A, B, C, D, E, G, and H. In subgroup F1, some African strains clustered with previously described strains from Brazil and Romania, suggesting an African origin of the HIV-1 epidemic in these countries. A more detailed analysis of the gag and the envelope sequences allowed the identification of four recombinant viruses. Our data show a high diversity among subtype F strains, suggesting the presence of new subtypes in the regions studied. If biological differences exist among subtypes, it is important that these subtypes be well defined. The data from our study show that there is a need to clearly identify the different subgroups within the F subtype. PMID- 10364494 TI - Alterations in RANTES gene expression and T-cell prevalence in intestinal mucosa during pathogenic or nonpathogenic simian immunodeficiency virus infection. AB - RANTES, a beta-chemokine, can suppress human immunodeficiency virus (HIV) as well as simian immunodeficiency virus (SIV) infections in T-lymphocyte cultures in vitro. However, the association of RANTES levels in peripheral blood with viral loads and disease outcome in HIV infection has been inconclusive. SIV-infected rhesus macaques were evaluated to determine whether RANTES gene expression correlated with suppression of viral infection in intestinal lymphoid tissues. Intestinal tissues were obtained from rhesus macaques infected with either pathogenic or nonpathogenic SIVmac variants at various stages of infection (primary acute, asymptomatic, and terminal). We examined the level of SIV infection (in situ hybridization), RANTES expression (quantitative competitive RT PCR), and T-cell counts (immunohistochemistry). The most pronounced increase in RANTES gene expression in intestinal tissues was observed in primary SIV infection, which correlated with the pathogenicity of the infecting virus and not the tissue viral loads. Our results demonstrated that in contrast to the occurrence of viral suppression by RANTES in vitro, there was no direct correlation between high RANTES gene expression and suppression of viral loads in intestinal lymphoid tissues. Thus RANTES expression in the gut lymphoid tissue may not be a correlate for viral suppression. However, RANTES gene expression in primary SIV infection may be part of early host immune response to viral infection. PMID- 10364495 TI - Novel expression of mouse adenovirus type 1 early region 3 gp11K at late times after infection. AB - Mutations were introduced into mouse adenovirus type 1 (MAV-1) early region 3 (E3) initiator codons by homologous recombination between viral DNA and a plasmid containing a mutagenized E3 region. The resulting mutant virus, pmE312, contained ATG --> TTA mutations at codon positions 1 and 4 and was expected to be null for the expression of the E3 proteins. However, gp11K, an MAV-1 E3 glycoprotein of 14K molecular weight, was detected in mutant-infected cell lysates at levels about 10-12% of that of wild-type (wt) virus at late times in infection. The gp11K polypeptide produced by pmE312 at late times was immunoprecipitated with two E3-specific antisera prepared against different regions of the protein. Like gp11K produced by wt virus infections, it was sensitive to endoglycosidase H (endo H) and thus resident in the endoplasmic reticulum (ER). In pmE312-infected cells treated with cytosine arabinoside (araC), an inhibitor of DNA replication, the gp11K protein was not detected by immunoprecipitation. This indicates that gp11K expression in pmE312-infected cells at late times was dependent on DNA replication and that it was thus translated from a late transcript. In vitro translation of poly(A)+ RNA from mock-, wild-type-, and pmE312-infected cells showed that gp11K was translated from late mRNA as an approximately 28K fusion between a late protein and gp11K. Our data are consistent with a model in which gp11K is expressed at late times as a late protein-gp11K chimera in both wt- and mutant-infected cells. This chimera is then processed: removal of a large N terminal sequence results in the observed 14K ER-localized gp11K. PMID- 10364496 TI - Recombinant viruses expressing the foot-and-mouth disease virus capsid precursor polypeptide (P1) induce cellular but not humoral antiviral immunity and partial protection in pigs. AB - The importance of the induction of virus neutralizing antibodies to provide protection against foot-and-mouth disease virus (FMDV) infection is well established. However, recent studies with recombinant adenovirus expressing the precursor polypeptide of the viral capsid (P1) indicate that cattle inoculated with this recombinant vector developed partial protection against FMDV infection, in the absence of a detectable specific humoral response. Other viral vectors have been widely used to induce protective immunity against many pathogens, and it has been reported that the use of different vectors for priming and boosting injections can provide a synergistic effect on this response. In this work, we determined the immunogenicity of two recombinant viruses (adenovirus and vaccinia) expressing P1-FMDV, administered either individually or sequentially, and the protection that they induced against FMDV challenge in pigs. A double immunization with the adeno-P1 virus was the most effective strategy at inducing protective immunity. In contrast to previous reports, the use of two different vectors for priming and boosting did not show a synergistic effect on the protection induced against FMD. Interestingly, immunized pigs developed FMDV specific T cell responses but not detectable antibodies. Thus, the protection observed was likely to be mediated by a cellular immune response. PMID- 10364497 TI - High-yield reassortant influenza vaccine production virus has a mutation at an HLA-A 2.1-restricted CD8+ CTL epitope on the NS1 protein. AB - Current influenza virus vaccines are prepared using high-yield reassortant virus strains obtained from a mixed infection of the new virus strain and a prototype high-yielding virus strain. The high-titered reassortant virus strain used as vaccine seed virus possesses the recent virus HA and NA and contains the internal genes from the high-growing prototype parent. We established a human CD8(+) cytotoxic T cell (CTL) line, 10-2C2, which recognizes an HLA-A2.1-restricted influenza A virus H1, H2, H3 cross-reactive T cell epitope on amino acids 122-130 of the NS1 protein, and unexpectedly we observed that there was decreased lysis of target cells infected with the A/Texas/36/91 (H1N1) vaccine virus strain compared to the lysis of target cells infected with the prototype A/PR/8/34 (H1N1) virus. RT-PCR results showed that the A/Texas vaccine virus strain contained a quasispecies. Approximately 50% of viral RNA of the NS1 gene had a nucleotide substitution that resulted in the N --> K amino acid change at the sixth position of the nonamer peptide. Current influenza vaccines are inactivated and do not contain the NS1 protein; however, future influenza vaccines may include live attenuated vaccines and with this mutation a live virus would fail to induce a CD8(+) CTL response to this epitope in individuals with HLA-A2.1, a very common allele, and potentially have reduced efficacy. PMID- 10364498 TI - Role of the cytoplasmic tail of gE in antibody-induced redistribution of viral glycoproteins expressed on pseudorabies-virus-infected cells. AB - Pseudorabies virus (PrV) glycoprotein gE is a nonessential glycoprotein involved in virulence and spread of the virus. It also has an important, yet unknown, function during antibody-induced capping of viral glycoproteins on the plasma membrane of PrV-infected swine kidney cells. In the present study, it was shown, by the use of a PrV strain expressing a truncated gE glycoprotein, that the cytoplasmic tail of gE is of significant importance for viral glycoprotein capping to occur. In addition, using PrV strains carrying point mutations in the cytoplasmic tail of gE, it was demonstrated that two tyrosine-based motifs are very important for correct functioning of gE during viral glycoprotein capping. Furthermore it was shown that genistein and tyrphostin, two tyrosine kinase activity inhibitors, inhibit viral glycoprotein capping in a concentration dependent manner. In conclusion, it can be stated that efficient antibody-induced viral glycoprotein capping requires the presence of two YxxL sequences in the cytoplasmic tail of glycoprotein gE, as well as the activation of a tyrosine phosphorylation signal transduction pathway. PMID- 10364499 TI - Immune responses and protection obtained by oral immunization with rotavirus VP4 and VP7 DNA vaccines encapsulated in microparticles. AB - Protective immune responses in mice were obtained after oral immunization with rotavirus DNA vaccines encapsulated in poly(lactide-co-glycolide) (PLG) microparticles. The DNA vaccines used encoded outer capsid proteins VP4 and VP7; proteins that are the basis for rotavirus serotyping and the generation of virus neutralizing antibodies. One dose of vaccine was given to BALB/c mice by oral gavage (75 microg DNA/mouse). Rotavirus-specific serum antibodies and intestinal IgA antibodies were detectable by 6 weeks postimmunization. After challenge with homologous murine rotavirus at 12 weeks postimmunization, fecal rotavirus antigen was reduced significantly in immunized mice compared with controls. Protective immunity also was generated by oral delivery of unencapsulated VP 7 DNA vaccine but to a lesser degree. These results demonstrate that the oral route is effective for generating protective immune responses with rotavirus DNA vaccines targeting neutralization antigens. PMID- 10364500 TI - Higher selection pressure from antiretroviral drugs in vivo results in increased evolutionary distance in HIV-1 pol. AB - We investigated the effect of selection pressures on evolution of HIV-1 pol in 51 patients after switching to a new antiretroviral combination reverse transcriptase (RT) inhibitor therapy. Evolution of the protease (PR) and RT reading frames were analysed separately. Pairwise evolutionary distances (ED) were calculated between sequences from baseline and week 8 and between baseline and week 48 of protocol therapy. ED were calculated for all substitutions and for synonymous and nonsynonymous substitutions separately. At week 8 when HIV RNA reduction (selection pressure) was high, significantly more divergence in pol in both synonymous and nonsynonymous substitutions was found in patients with substantial RNA reduction (strong responders). Separate analyses of PR and RT revealed significantly greater ED in the RT (under selection pressure) of strong compared with nonresponders, whereas divergence between PR genes (not under selection pressure) did not differ in those two groups. Such differential evolution indicates that PR and RT were genetically unlinked and suggests recombination. The rapid increase of ED over the first 8 weeks was followed by only a minimal further rise by week 48, suggesting that selection of preexisting quasispecies accounted for the early changes. A disproportionally high number of synonymous substitutions accounted for the observed divergence and indicated that such genetic changes may not be completely silent. PMID- 10364501 TI - Pathogenicity and comparative evolution in vivo of the transitional quasispecies SIVsmmPBj8. AB - During 14 months of infection of a pig-tailed macaque, the acutely lethal simian immunodeficiency virus SIVsmmPBj14 (SIV-PBj14) evolved from the minimally pathogenic strain SIVsmm9. The virus isolated at 8 months (SIV-PBj8) exhibited properties of both SIVsmm9 and SIV-PBj14, indicating that a phenotypic transition occurred between 6 and 10 months. To assess the influence that this new composition of biologic properties might have on pathogenicity, three pig-tailed macaques were inoculated intravenously with SIV-PBj8. Although no animals developed the severe acute disease syndrome typical of SIV-PBj14, all had high levels of viremia and died of AIDS at 4, 10. 5, and 32 months. Characterization of the SIV-PBj8-derived quasispecies that evolved in these macaques showed that at 4 days after inoculation, viruses from all three animals exhibited in vitro properties different from those of the inoculum. By 4 months, the initial phenotypic profiles had changed, with the quasispecies in plasma from the animal (J90232) that died at this time most closely resembling SIV-PBj14, not SIV-PBj8. Phylogenetic trees of the gp41/Nef region of viruses in 4-month plasma from J90232 revealed three distinct populations with high bootstrap values: one group branched with SIVsmm9, one with SIV-PBj14, and one with SIV-PBj8 (ratio of clones, 5:9:5). Nucleotide sequence analysis suggested that some members of the original SIV-PBj8 quasispecies may have been evolving toward a SIV-PBj14-like genotype at the time macaque J90232 died. The use of SIV-PBj8, which was more pathogenic than SIVsmm9, but less pathogenic than SIV-PBj14, may provide the optimal genetic background on which to identify the minimal, multigenic determinants of the SIV-PBj14 phenotype. The results of our studies on SIV-PBj14 indicate that in some, but not all, cases of primate lentivirus infection more pathogenic variants evolve, selectively proliferate, and more than likely contribute to disease progression. PMID- 10364502 TI - Catalytic features of the recombinant reverse transcriptase of bovine leukemia virus expressed in bacteria. AB - We have expressed the recombinant reverse transcriptase (RT) of bovine leukemia virus (BLV) in bacteria. The gene encoding the RT was designed to start at its 5' end next to the last codon of the mature viral protease, namely the amino terminus of the RT matches the last 26 codons of the pro gene and is coded for by the pro reading frame. The RT sequence extends into the pol gene, utilizing the pol reading frame after overcoming the stop codon by adding an extra nucleotide (thus imitating the naturally occurring frameshift event). Hence we have generated a transframe polypeptide that is a 584-residues-long protein (see Rice, Stephens, Burny, and Gilden (1985) Virology 142, 357-377). This protein was partially purified after adding a six-histidine tag and studied biochemically testing a variety of parameters. The enzyme exhibits all activities typical of RTs, i.e., both RNA- and DNA-dependent DNA polymerase as well as a ribonuclease H (RNase H) activity. Unlike most RTs, the BLV RT is enzymatically active as a monomer even after binding a DNA substrate. The enzyme shows a preference for Mg2+ over Mn2+ in both its DNA polymerase and RNase H activities. BLV RT is relatively resistant to nucleoside triphosphate analogues, which are known to be potent inhibitors of other RTs such as that of HIV. PMID- 10364503 TI - Identification of a novel circular single-stranded DNA associated with cotton leaf curl disease in Pakistan. AB - Recent reports have suggested that cotton leaf curl virus (CLCuV), a geminivirus of the genus Begomovirus, may be responsible for cotton leaf curl disease in Pakistan. However, the causal agent of the disease remains unclear as CLCuV genomic components resembling begomovirus DNA A are unable to induce typical disease symptoms when reintroduced into plants. All attempts to isolate a genomic component equivalent to begomovirus DNA B have been unsuccessful. Here, we describe the isolation and characterisation of a novel circular single-stranded (ss) DNA associated with naturally infected cotton plants. In addition to a component resembling DNA A, purified geminate particles contain a smaller unrelated ssDNA that we refer to as DNA 1. DNA 1 was cloned from double-stranded replicative form of the viral DNA isolated from infected cotton plants. Blot hybridisation using probes specific for either CLCuV DNA or DNA 1 was used to demonstrate that both DNAs co-infect naturally infected cotton plants from different geographical locations. DNA 1 was detected in viruliferous Bemisia tabaci and in tobacco plants infected under laboratory conditions using B. tabaci, indicating that it is transmitted by whiteflies. Sequence analysis showed that DNA 1 is approximately half the size of CLCuV DNA but shares no homology, indicating that it is not a defective geminivirus component. DNA 1 has some homology to a genomic component of members of Nanoviridae, a family of DNA viruses that are normally transmitted by aphids or planthoppers. DNA 1 encodes a homologue of the nanovirus replication-associated protein (Rep) and has the capacity to autonomously replicate in tobacco. The data suggest that a nanovirus like DNA has become whitefly-transmissible as a result of its association with a geminivirus and that cotton leaf curl disease may result from a mutually dependent relationship that has developed between members of two distinct DNA virus families that share a similar replication strategy. PMID- 10364504 TI - The N-terminal half of the brome mosaic virus 1a protein has RNA capping associated activities: specificity for GTP and S-adenosylmethionine. AB - The N-terminal half of the brome mosaic virus (BMV) 1a replication-associated protein contains sequence motifs found in RNA methyltransferases. We demonstrate that recombinant BMV methyltransferase-like (MT) domain expressed in Escherichia coli forms an adduct with a guanine nucleotide in a reaction that requires S adenosylmethionine (AdoMet) and divalent cations. Moieties in GTP and AdoMet required for adduct formation were determined using a competition assay and chemical analogues. In the guanine nucleotide the ribose 2' hydroxyl, the triphosphates, the base C6 keto group, and possibly the N1 imine are required. In AdoMet, the methyl group and the ability to transfer a methyl group to guanine nucleotide were demonstrated to be required for adduct formation. The effects of methyltransferase inhibitors on viral RNA synthesis was determined using an in vitro RNA synthesis assay. These results are consistent with the previously reported activities of alphaviral nsP1 methyltransferase protein and identify the chemical moieties required for the BMV methyltransferase activity. PMID- 10364505 TI - Nucleotides 1506-1625 of bovine papillomavirus type 1 genome can enhance DNA packaging by L1/L2 capsids. AB - We have previously described a DNA-packaging assay using bovine papillomavirus type 1 (BPV-1) virus-like particles (VLPs) and have identified a region of the BPV genome that assists in packaging. In this study, we identify a specific BPV sequence involved in DNA packaging by BPV-1 VLPs. In the initial screening of BPV 1 genomic sequences essential for DNA packaging, we observed that a plasmid with deletions between nucleotides (nt) 948 and 2113 failed to be packaged into BPV-1 VLPs. However, plasmids containing nt 948 to 2113 were efficiently packaged, suggesting that this 1.2-kb fragment contains a packaging enhancement sequence (PES). Further mapping of the BPV-1 genome showed that this packaging sequence lies between nt 1506 and 1625. Furthermore, this packaging sequence is also recognized by HPV6b VLPs, suggesting that a common packaging mechanism may be used by the two papillomavirus types. Given the phylogenetic difference between these two viral types, it is likely that other papillomavirus types may also use the same packaging mechanism. Identification of the PES has allowed a minimal viral genome sequence to be used in the packaging assay, improving the usefulness of the assay in studying the process of papillomavirus DNA encapsidation. PMID- 10364506 TI - Carboxy-terminal five amino acids of the nucleocapsid protein of vesicular stomatitis virus are required for encapsidation and replication of genome RNA. AB - The encapsidation of vesicular stomatitis virus (VSV) genome RNA, a prerequisite step to the replication process by the nucleocapsid protein (N) was studied by its ability to package VSV leader RNA in vitro in a RNase-resistant form. The VSV leader RNA was derived from the SP6 transcription vector while the N protein was made in rabbit reticulocyte lysate. The in vitro encapsidation was carried out by translating N mRNA in the presence of 32P-labeled presynthesized leader RNA. The RNA encapsidation property of the N protein was completely abrogated when the C terminal five amino acids (VEFDK-COOH) were deleted. Systematic mutational analyses within the C-terminal five amino acid regions reveal that the RNA encapsidation activity was lost in all mutants except K --> A and K --> R, indicating that C-terminal five amino acids, in particular the lysine residue play critical role in genome RNA encapsidation. To correlate the in vitro encapsidation abilities of these mutant N proteins with genome RNA replication, we have used a full-length cDNA clone of VSV genome RNA to rescue infectious virions from cells expressing L, P, and wt or mutant N proteins and measured the recovery of plaque forming units. The results indicate that the N mutants that are defective in in vitro encapsidation of leader RNA do not support replication, establishing the requirement of C-terminal five amino acids of the N protein in viral replication. PMID- 10364507 TI - SVMPA, a mutant of sindbis virus resistant to mycophenolic acid and ribavirin, shows an increased sensitivity to chick interferon. AB - SVMPA is a mutant of Sindbis virus, selected for its ability to replicate in mycophenolic acid (MPA)-treated mosquito cells. SVMPA has another phenotype: although able to replicate normally in primary cultures of chick embryo fibroblasts (CEF), its replication is restricted in secondary cultures prepared from aged primary CEF cultures. The mutations responsible for these phenotypes mapped to the region of the viral genome that codes for nsP1. We report here that SVMPA has yet another phenotype. Relative to our standard Sindbis virus (SVSTD) from which it was derived, SVMPA shows an increased sensitivity to chick interferon, both crude interferon prepared from virus-infected cells and recombinant interferon. Characterization of viral mutants obtained after site directed mutagenesis indicated that the same mutations responsible for the host restriction of SVMPA in secondary cultures of CEF were also responsible for its increased sensitivity to chick interferon. PMID- 10364508 TI - Symptom attenuation by a satellite RNA in vivo is dependent on reduced levels of virus coat protein. AB - Many plant RNA viruses provide replication and encapsidation functions for one or more satellite RNAs (sat-RNAs) that can modulate the symptoms of the associated helper virus. Sat-RNA C, a virulent sat-RNA associated with turnip crinkle virus (TCV), normally intensifies symptoms but can attenuate symptoms if the TCV coat protein (CP) is replaced with that of cardamine chlorotic fleck carmovirus [Kong et al. (1995) Plant Cell 7, 1625-1634] or if TCV contains an alteration in the CP initiation codon (TCV-CPm) [Kong et al. (1997b) Plant Cell 9, 2051-2063]. To further elucidate the mechanism of symptom attenuation by sat-RNA C, the composition of the CP produced by TCV-CPm (CPCPm) was determined. Our results reveal that CPCPm likely has two additional amino acids at its N-terminus compared with wild-type TCV CP. TCV-CPm produces reduced levels of CP, and this reduction, not the two additional residues at the CP N-terminus, is responsible for symptom attenuation by sat-RNA C. PMID- 10364509 TI - Genetics of Angelman syndrome. PMID- 10364511 TI - Microglia and the immune pathology of Alzheimer disease. PMID- 10364510 TI - Function and dysfunction of the presenilins. PMID- 10364512 TI - Periventricular heterotopia and the genetics of neuronal migration in the cerebral cortex. PMID- 10364513 TI - Learning from the slime mold: Dictyostelium and human disease. PMID- 10364514 TI - Identification of novel pro-alpha2(IX) collagen gene mutations in two families with distinctive oligo-epiphyseal forms of multiple epiphyseal dysplasia. AB - Multiple epiphyseal dysplasia (MED) is a genetically heterogeneous disorder with marked clinical and radiographic variability. Traditionally, the mild "Ribbing" and severe "Fairbank" types have been used to define a broad phenotypic spectrum. Mutations in the gene encoding cartilage oligomeric-matrix protein have been shown to result in several types of MED, whereas mutations in the gene encoding the alpha2 chain of type IX collagen (COL9A2) have so far been found only in two families with the Fairbank type of MED. Type IX collagen is a heterotrimer of pro alpha chains derived from three distinct genes-COL9A1, COL9A2, and COL9A3. In this article, we describe two families with distinctive oligo-epiphyseal forms of MED, which are heterozygous for different mutations in the COL9A2 exon 3/intron 3 splice-donor site. Both of these mutations result in the skipping of exon 3 from COL9A2 mRNA, but the position of the mutation in the splice-donor site determines the stability of the mRNA produced from the mutant COL9A2 allele. PMID- 10364515 TI - Identification of mutations in the repeated part of the autosomal dominant polycystic kidney disease type 1 gene, PKD1, by long-range PCR. AB - We have used long-range PCR to identify mutations in the duplicated part of the PKD1 gene. By means of a PKD1-specific primer in intron 1, an approximately 13.6 kb PCR product that includes exons 2-15 of the PKD1 gene has been used to search for mutations, by direct sequence analysis. This region contains the majority of the predicted extracellular domains of the PKD1-gene product, polycystin, including the 16 novel PKD domains that have similarity to immunoglobulin-like domains found in many cell-adhesion molecules and cell-surface receptors. Direct sequence analysis of exons encoding all the 16 PKD domains was performed on PCR products from a group of 24 unrelated patients with autosomal dominant polycystic kidney disease (ADPKD [MIM 173900]). Seven novel mutations were found in a screening of 42% of the PKD1-coding region in each patient, representing a 29% detection rate; these mutations included two deletions (one of 3 kb and the other of 28 bp), one single-base insertion, and four nucleotide substitutions (one splice site, one nonsense, and two missense). Five of these mutations would be predicted to cause a prematurely truncated protein. Two coding and 18 silent polymorphisms were also found. When, for the PKD1 gene, this method is coupled with existing mutation-detection methods, virtually the whole of this large, complex gene can now be screened for mutations. PMID- 10364516 TI - X-linked dyskeratosis congenita is predominantly caused by missense mutations in the DKC1 gene. AB - Dyskeratosis congenita is a rare inherited bone marrow-failure syndrome characterized by abnormal skin pigmentation, nail dystrophy, and mucosal leukoplakia. More than 80% of patients develop bone-marrow failure, and this is the major cause of premature death. The X-linked form of the disease (MIM 305000) has been shown to be caused by mutations in the DKC1 gene. The gene encodes a 514 amino-acid protein, dyskerin, that is homologous to Saccharomyces cerevisiae Cbf5p and rat Nap57 proteins. By analogy to the homologues in other species, dyskerin is predicted to be a nucleolar protein with a role in both the biogenesis of ribosomes and, in particular, the pseudouridylation of rRNA precursors. We have determined the genomic structure of the DKC1 gene; it consists of 15 exons spanning a region of 15 kb. This has enabled us to screen for mutations in the genomic DNA, by using SSCP analysis. Mutations were detected in 21 of 37 additional families with dyskeratosis congenita that were analyzed. These mutations consisted of 11 different single-nucleotide substitutions, which resulted in 10 missense mutations and 1 putative splicing mutation within an intron. The missense change A353V was observed in 10 different families and was shown to be a recurring de novo event. Two polymorphisms were also detected, one of which resulted in the insertion of an additional lysine in the carboxy terminal polylysine domain. It is apparent that X-linked dyskeratosis congenita is predominantly caused by missense mutations; the precise effect on the function of dyskerin remains to be determined. PMID- 10364517 TI - The molecular basis of cystathionine beta-synthase deficiency in Dutch patients with homocystinuria: effect of CBS genotype on biochemical and clinical phenotype and on response to treatment. AB - Homocystinuria due to cystathionine beta-synthase (CBS) deficiency, inherited as an autosomal recessive trait, is the most prevalent inborn error of methionine metabolism. Its diverse clinical expression may include ectopia lentis, skeletal abnormalities, mental retardation, and premature arteriosclerosis and thrombosis. This variability is likely caused by considerable genetic heterogeneity. We investigated the molecular basis of CBS deficiency in 29 Dutch patients from 21 unrelated pedigrees and studied the possibility of a genotype-phenotype relationship with regard to biochemical and clinical expression and response to homocysteine-lowering treatment. Clinical symptoms and biochemical parameters were recorded at diagnosis and during long-term follow-up. Of 10 different mutations detected in the CBS gene, 833T-->C (I278T) was predominant, present in 23 (55%) of 42 independent alleles. At diagnosis, homozygotes for this mutation (n=12) tended to have higher homocysteine levels than those seen in patients with other genotypes (n=17), but similar clinical manifestations. During follow-up, I278T homozygotes responded more efficiently to homocysteine-lowering treatment. After 378 patient-years of treatment, only 2 vascular events were recorded; without treatment, at least 30 would have been expected (P<.01). This intervention in Dutch patients significantly reduces the risk of cardiovascular disease and other sequelae of classical homocystinuria syndrome. PMID- 10364518 TI - Characterization of a germline mosaicism in families with Lowe syndrome, and identification of seven novel mutations in the OCRL1 gene. AB - The oculocerebrorenal syndrome of Lowe (OCRL) is an X-linked disorder characterized by major abnormalities of eyes, nervous system, and kidneys. Mutations in the OCRL1 gene have been associated with the disease. OCRL1 encodes a phosphatidylinositol 4, 5-biphosphate (PtdIns[4,5]P2) 5-phosphatase. We have examined the OCRL1 gene in eight unrelated patients with OCRL and have found seven new mutations and one recurrent in-frame deletion. Among the new mutations, two nonsense mutations (R317X and E558X) and three other frameshift mutations caused premature termination of the protein. A missense mutation, R483G, was located in the highly conserved PtdIns(4,5)P2 5-phosphatase domain. Finally, one frameshift mutation, 2799delC, modifies the C-terminal part of OCRL1, with an extension of six amino acids. Altogether, 70% of missense mutations are located in exon 15, and 52% of all mutations cluster in exons 11-15. We also identified two new microsatellite markers for the OCRL1 locus, and we detected a germline mosaicism in one family. This observation has direct implications for genetic counseling of Lowe syndrome families. PMID- 10364519 TI - Structure of the GM2A gene: identification of an exon 2 nonsense mutation and a naturally occurring transcript with an in-frame deletion of exon 2. AB - Deficiency of the GM2 activator protein, encoded by GM2A, results in the rare AB variant form of GM2 gangliosidosis. Four mutations have been identified, but the human gene structure has been only partially characterized. We report a new patient from a Laotian deme whose cells are deficient in both GM2-activator mRNA and protein. However, reverse transcription (RT)-PCR detected some normal-sized cDNA and a smaller cDNA species, which was not seen in the RT-PCR products from normal controls. Sequencing revealed that, although the patient's normal-sized cDNA contained a single nonsense mutation in exon 2, his smaller cDNA was the result of an in-frame deletion of exon 2. Long PCR was used to amplify introns 1 and 2 from patient and normal genomic DNA, and no differences in size, in 5' and 3' end sequences, or in restriction-mapping patterns were observed. From these data we developed a set of four PCR primers that can be used to identify GM2A mutations. We use this procedure to demonstrate that the patient is likely homozygous for the nonsense mutation. Other reports have associated nonsense mutations with dramatically reduced levels of mRNA and with an increased level of skipping of the exon containing the mutation, thus reestablishing an open reading frame. However, a recent article has concluded that, in some cases, the latter observation is caused by an artifact of RT-PCR. In support of this conclusion, we demonstrate that, if the competing, normal-sized cDNA is removed from the initial RT-PCR products, from both patient and normal cells, by an exon 2-specific restriction digest; a second round of PCR produces similar amounts of exon 2 deleted cDNA. PMID- 10364520 TI - MEFV-Gene analysis in armenian patients with Familial Mediterranean fever: diagnostic value and unfavorable renal prognosis of the M694V homozygous genotype genetic and therapeutic implications. AB - Familial Mediterranean fever (FMF) is a recessively inherited disorder that is common in patients of Armenian ancestry. To date, its diagnosis, which can be made only retrospectively, is one of exclusion, based entirely on nonspecific clinical signs that result from serosal inflammation and that may lead to unnecessary surgery. Renal amyloidosis, prevented by colchicine, is the most severe complication of FMF, a disorder associated with mutations in the MEFV gene. To evaluate the diagnostic and prognostic value of MEFV-gene analysis, we investigated 90 Armenian FMF patients from 77 unrelated families that were not selected through genetic-linkage analysis. Eight mutations, one of which (R408Q) is new, were found to account for 93% of the 163 independent FMF alleles, with both FMF alleles identified in 89% of the patients. In several instances, family studies provided molecular evidence for pseudodominant transmission and incomplete penetrance of the disease phenotype. The M694V homozygous genotype was found to be associated with a higher prevalence of renal amyloidosis and arthritis, compared with other genotypes (P=.0002 and P=.006, respectively). The demonstration of both the diagnostic and prognostic value of MEFV analysis and particular modes of inheritance should lead to new ways for management of FMF including genetic counseling and therapeutic decisions in affected families. PMID- 10364521 TI - Noninvasive test for fragile X syndrome, using hair root analysis. AB - Identification of the FMR1 gene and the repeat-amplification mechanism causing fragile X syndrome led to development of reliable DNA-based diagnostic methods, including Southern blot hybridization and PCR. Both methods are performed on DNA isolated from peripheral blood cells and measure the repeat size in FMR1. Using an immunocytochemical technique on blood smears, we recently developed a novel test for identification of patients with fragile X syndrome. This method, also called "antibody test," uses monoclonal antibodies against the FMR1 gene product (FMRP) and is based on absence of FMRP in patients' cells. Here we describe a new diagnostic test to identify male patients with fragile X syndrome, on the basis of lack of FMRP in their hair roots. Expression of FMRP in hair roots was studied by use of an FMRP-specific antibody test, and the percentage of FMRP-expressing hair roots in controls and in male fragile X patients was determined. Control individuals showed clear expression of FMRP in nearly every hair root, whereas male fragile X patients lacked expression of FMRP in almost all their hair roots. Mentally retarded female patients with a full mutation showed FMRP expression in only some of their hair roots (<55%), and no overlap with normal female controls was observed. The advantages of this test are (1) plucking of hair follicles does no appreciable harm to the mentally retarded patient, (2) hairs can be sent in a simple envelope to a diagnostic center, and (3) the result of the test is available within 5 h of plucking. In addition, this test enabled us to identify two fragile X patients who did not show the full mutation by analysis of DNA isolated from blood cells. PMID- 10364522 TI - Monodactylous limbs and abnormal genitalia are associated with hemizygosity for the human 2q31 region that includes the HOXD cluster. AB - Vertebrates have four clusters of Hox genes (HoxA, HoxB, HoxC, and HoxD). A variety of expression and mutation studies indicate that posterior members of the HoxA and HoxD clusters play an important role in vertebrate limb development. In humans, mutations in HOXD13 have been associated with type II syndactyly or synpolydactyly, and, in HOXA13, with hand-foot-genital syndrome. We have investigated two unrelated children with a previously unreported pattern of severe developmental defects on the anterior-posterior (a-p) limb axis and in the genitalia, consisting of a single bone in the zeugopod, either monodactyly or oligodactyly in the autopod of all four limbs, and penoscrotal hypoplasia. Both children are heterozygous for a deletion that eliminates at least eight (HOXD3 HOXD13) of the nine genes in the HOXD cluster. We propose that the patients' phenotypes are due in part to haploinsufficiency for HOXD-cluster genes. This hypothesis is supported by the expression patterns of these genes in early vertebrate embryos. However, the involvement of additional genes in the region could explain the discordance, in severity, between these human phenotypes and the milder, non-polarized phenotypes present in mice hemizygous for HoxD cluster genes. These cases represent the first reported examples of deficiencies for an entire Hox cluster in vertebrates and suggest that the diploid dose of human HOXD genes is crucial for normal growth and patterning of the limbs along the anterior posterior axis. PMID- 10364524 TI - A gene for inherited cutaneous venous anomalies ("glomangiomas") localizes to chromosome 1p21-22. AB - Venous malformations (VMs) are localized defects of vascular morphogenesis. They can occur in every organ system, most commonly in skin and muscle. They can cause pain and bleeding, and in some critical locations they can be life threatening. Usually venous anomalies occur sporadically, but families with dominant inheritance have been identified. Using linkage analysis, we have established in earlier reports that some families with inherited VMs show linkage to chromosome 9p21; the mutation causes ligand-independent activation of an endothelial cell specific receptor tyrosine kinase, TIE-2. Here we show that VMs with glomus cells (known as "glomangiomas"), inherited as an autosomal dominant trait in five families, are not linked to 9p21 but, instead, link to a new locus, on 1p21-p22, called "VMGLOM" (LOD score 12.70 at recombination fraction.00). We exclude three known positional candidate genes, DR1 (depressor of transcription 1), TGFBR3 (transforming growth factor-beta receptor, type 3), and TFA (tissue factor). We hypothesize that cutaneous venous anomalies (i.e., glomangiomas) are caused by mutations in a novel gene that may act to regulate angiogenesis, in concert with the TIE-2 signaling pathway. PMID- 10364523 TI - Campomelic dysplasia translocation breakpoints are scattered over 1 Mb proximal to SOX9: evidence for an extended control region. AB - Campomelic dysplasia (CD), a skeletal malformation syndrome with or without XY sex reversal, is usually caused by mutations within the SOX9 gene on distal 17q. Several CD translocation and inversion cases have been described with breakpoints outside the coding region, mapping to locations >130 kb proximal to SOX9. Such cases are generally less severely affected than cases with SOX9 coding-region mutations, as is borne out by three new translocation cases that we present. We have cloned the region extending 1.2 Mb upstream of the SOX9 gene in overlapping bacterial-artificial-chromosome and P1-artificial-chromosome clones and have established a restriction map with rare-cutter enzymes. With sequence-tagged-site content mapping in somatic-cell hybrids, as well as with FISH, we have precisely mapped the breakpoints of the three new and of three previously described CD cases. The six CD breakpoints map to an interval that is 140-950 kb proximal to the SOX9 gene. With exon trapping, we could isolate five potential exons from the YAC 946E12 that spans the region, four of which could be placed in the contig in the vicinity of the breakpoints. They show the same transcriptional orientation, but only two have an open reading frame (ORF). We failed to detect expression of these fragments in several human and mouse cDNA libraries, as well as on northern blots. Genomic sequence totaling 1,063 kb from the SOX9 5'-flanking region was determined and was analyzed by the gene-prediction program GENSCAN and by a search of dbEST and other databases. No genes or transcripts could be identified. Together, these data suggest that the chromosomal rearrangements most likely remove one or more cis-regulatory elements from an extended SOX9 control region. PMID- 10364525 TI - In Swedish families with hereditary prostate cancer, linkage to the HPC1 locus on chromosome 1q24-25 is restricted to families with early-onset prostate cancer. AB - Prostate cancer clusters in some families, and an estimated 5%-10% of all cases are estimated to result from inheritance of prostate cancer-susceptibility genes. We previously reported evidence of linkage to the 1q24-25 region (HPC1) in 91 North American and Swedish families each with multiple cases of prostate cancer (Smith et al. 1996). In the present report we analyze 40 (12 original and 28 newly identified) Swedish families with hereditary prostate cancer (HPC) that, on the basis of 40 markers spanning a 25-cM interval within 1q24-25, have evidence of linkage. In the complete set of families, a maximum two-point LOD score of 1.10 was observed at D1S413 (at a recombination fraction [theta] of.1), with a maximum NPL (nonparametric linkage) Z score of 1.64 at D1S202 (P=.05). The evidence of linkage to this region originated almost exclusively from the subset of 12 early-onset (age <65 years) families, which yielded a maximum LOD score of 2.38 at D1S413 (straight theta=0) and an NPL Z score of 1.95 at D1S422 (P=.03). Estimates from heterogeneity tests suggest that, within Sweden, as many as 50% of early-onset families had evidence of linkage to the HPC1 region. These results are consistent with the hypothesis of linkage to HPC1 in a subset of families with prostate cancer, particularly those with an early age at diagnosis. PMID- 10364526 TI - A gene for fluctuating, progressive autosomal dominant nonsyndromic hearing loss, DFNA16, maps to chromosome 2q23-24.3. AB - The sixteenth gene to cause autosomal dominant nonsyndromic hearing loss (ADNSHL), DFNA16, maps to chromosome 2q23-24.3 and is tightly linked to markers in the D2S2380-D2S335 interval. DFNA16 is unique in that it results in the only form of ADNSHL in which the phenotype includes rapidly progressing and fluctuating hearing loss that appears to respond to steroid therapy. This observation suggests that it may be possible to stabilize hearing through medical intervention, once the biophysiology of deafness due to DFNA16 is clarified. Especially intriguing is the localization of several voltage-gated sodium-channel genes to the DFNA16 interval. These cationic channels are excellent positional and functional DFNA16 candidate genes. PMID- 10364527 TI - The gene for cherubism maps to chromosome 4p16.3. AB - Cherubism is a rare familial disease of childhood characterized by proliferative lesions within the mandible and maxilla that lead to prominence of the lower face and an appearance reminiscent of the cherubs portrayed in Renaissance art. Resolution of these bony abnormalities is often observed after puberty. Many cases are inherited in an autosomal dominant fashion, although several cases without a family history have been reported. Using two families with clinically, radiologically, and/or histologically proved cherubism, we have performed a genomewide linkage search and have localized the gene to chromosome 4p16.3, with a maximum multipoint LOD score of 5. 64. Both families showed evidence of linkage to this locus. Critical meiotic recombinants place the gene in a 3-cM interval between D4S127 and 4p-telomere. Within this region a strong candidate is the gene for fibroblast growth factor receptor 3 (FGFR3); mutations in this gene have been implicated in a diverse set of disorders of bone development. PMID- 10364529 TI - Osteoarthritis-susceptibility locus on chromosome 11q, detected by linkage. AB - We present a two-stage genomewide scan for osteoarthritis-susceptibility loci, using 481 families that each contain at least one affected sibling pair. The first stage, with 272 microsatellite markers and 297 families, involved a sparse map covering 23 chromosomes at intervals of approximately 15 cM. Sixteen markers that showed evidence of linkage at nominal P/=40 CAG repeats) are generally most common in Asian populations, less common in populations of European ancestry, and least common among Africans. We have observed a high intergenerational instability (46. 3%+/-5.1%) of the large alleles. Although the mutation rate is not dependent on parental sex, paternal transmissions have predominantly resulted in contractions, whereas maternal transmissions have yielded expansions. Within this class of large alleles, the mutation rate increases concomitantly with increasing allele size, but the magnitude of repeat size change does not depend on the size of the progenitor allele. Sequencing of specific alleles reveals that the intermediate sized alleles (30-40 repeats) have CAT/CAC interruptions within the CAG-repeat array. These results indicate that expansion and instability of trinucleotide repeats are not exclusively disease-associated phenomena. The implications of the existence of massively expanded alleles in the general populations are not yet understood. PMID- 10364533 TI - An mtDNA analysis in ancient Basque populations: implications for haplogroup V as a marker for a major paleolithic expansion from southwestern europe. AB - mtDNA sequence variation was studied in 121 dental samples from four Basque prehistoric sites, by high-resolution RFLP analysis. The results of this study are corroborated by (1) parallel analysis of 92 bone samples, (2) the use of controls during extraction and amplification, and (3) typing by both positive and negative restriction of the linked sites that characterize each haplogroup. The absence of haplogroup V in the prehistoric samples analyzed conflicts with the hypothesis proposed by Torroni et al., in which haplogroup V is considered as an mtDNA marker for a major Paleolithic population expansion from southwestern Europe, occurring approximately 10,000-15,000 years before the present (YBP). Our samples from the Basque Country provide a valuable tool for checking the previous hypothesis, which is based on genetic data from present-day populations. In light of the available data, the most realistic scenario to explain the origin and distribution of haplogroup V suggests that the mutation defining that haplogroup (4577 NlaIII) appeared at a time when the effective population size was small enough to allow genetic drift to act-and that such drift is responsible for the heterogeneity observed in Basques, with regard to the frequency of haplogroup V (0%-20%). This is compatible with the attributed date for the origin of that mutation (10,000-15, 000 YBP), because during the postglacial period (the Mesolithic, approximately 11,000 YBP) there was a major demographic change in the Basque Country, which minimized the effect of genetic drift. This interpretation does not rely on migratory movements to explain the distribution of haplogroup V in present-day Indo-European populations. PMID- 10364535 TI - Use of unlinked genetic markers to detect population stratification in association studies. AB - We examine the issue of population stratification in association-mapping studies. In case-control studies of association, population subdivision or recent admixture of populations can lead to spurious associations between a phenotype and unlinked candidate loci. Using a model of sampling from a structured population, we show that if population stratification exists, it can be detected by use of unlinked marker loci. We show that the case-control-study design, using unrelated control individuals, is a valid approach for association mapping, provided that marker loci unlinked to the candidate locus are included in the study, to test for stratification. We suggest guidelines as to the number of unlinked marker loci to use. PMID- 10364536 TI - Methods for detection of parent-of-origin effects in genetic studies of case parents triads. AB - When affected probands and their biological parents are genotyped at a candidate gene or a marker, the resulting case-parents-triad data enable powerful tests for linkage in the presence of association. When linkage disequilibrium has been detected in such a study, the investigator may wish to look further for possible parent-of-origin effects. If, for example, the transmission/disequilibrium test restricted to fathers is statistically significant, whereas that restricted to mothers is not, the investigator might interpret this as evidence for nonexpression of the maternally derived disease gene-that is, imprinting. This report reviews existing methods for detection of parent-of-origin effects, showing that each can be invalid under certain scenarios. Two new methods are proposed, based on application of likelihood-based inference after stratification on both the parental mating type and the inherited number of copies of the allele under study. If there are no maternal genetic effects expressed prenatally during gestation, the parental-asymmetry test is powerful and provides valid estimation of a parent-of-origin parameter. For diseases for which there could be maternal effects on risk, the parent-of-origin likelihood-ratio test provides a robust alternative. Simulations based on an admixed population demonstrate good operating characteristics for these procedures, under diverse scenarios. PMID- 10364534 TI - Sex-specific migration patterns in Central Asian populations, revealed by analysis of Y-chromosome short tandem repeats and mtDNA. AB - Eight Y-linked short-tandem-repeat polymorphisms (DYS19, DYS388, DYS389I, DYS389II, DYS390, DYS391, DYS392, and DYS393) were analyzed in four populations of Central Asia, comprising two lowland samples-Uighurs and lowland Kirghiz-and two highland samples-namely, the Kazakhs (altitude 2,500 m above sea level) and highland Kirghiz (altitude 3,200 m above sea level). The results were compared with mtDNA sequence data on the same individuals, to study possible differences in male versus female genetic-variation patterns in these Central Asian populations. Analysis of molecular variance (AMOVA) showed a very high degree of genetic differentiation among the populations tested, in discordance with the results obtained with mtDNA sequences, which showed high homogeneity. Moreover, a dramatic reduction of the haplotype genetic diversity was observed in the villages at high altitude, especially in the highland Kirghiz, when compared with the villages at low altitude, which suggests a male founder effect in the settlement of high-altitude lands. Nonetheless, mtDNA genetic diversity in these highland populations is equivalent to that in the lowland populations. The present results suggest a very different migration pattern in males versus females, in an extended historical frame, with a higher migration rate for females. PMID- 10364537 TI - A test of transmission/disequilibrium for quantitative traits in pedigree data, by multiple regression. AB - The transmission/disequilibrium (TD) test (TDT), proposed, by Spielman et al., for binary traits is a powerful method for detection of linkage between a marker locus and a disease locus, in the presence of allelic association. As a test for linkage disequilibrium, the TDT makes the assumption that any allelic association present is due to linkage. Allison proposed a series of TD-type tests for quantitative traits and calculated their power, assuming that the marker locus is the disease locus. All these tests assume that the observations are independent, and therefore they are applicable, as a test for linkage, only for nuclear-family data. In this report, we propose a regression-based TD-type test for linkage between a marker locus and a quantitative trait locus, using information on the parent-to-offspring transmission status of the associated allele at the marker locus. This method does not require independence of observations, thus allowing for analysis of pedigree data as well, and allows adjustment for covariates. We investigate the statistical power and validity of the test by simulating markers at various recombination fractions from the disease locus. PMID- 10364538 TI - Mutations of UFD1L are not responsible for the majority of cases of DiGeorge Syndrome/velocardiofacial syndrome without deletions within chromosome 22q11. PMID- 10364539 TI - Haploinsufficiency of the HOXA gene cluster, in a patient with hand-foot-genital syndrome, velopharyngeal insufficiency, and persistent patent Ductus botalli. PMID- 10364541 TI - No evidence of linkage for chromosome 1q42.2-43 in prostate cancer. PMID- 10364542 TI - A third locus predisposing to multiple deletions of mtDNA in autosomal dominant progressive external ophthalmoplegia. PMID- 10364540 TI - Highly skewed X-chromosome inactivation is associated with idiopathic recurrent spontaneous abortion. PMID- 10364543 TI - Possible interaction between USH1B and USH3 gene products as implied by apparent digenic deafness inheritance. PMID- 10364544 TI - Anticipation in familial chronic lymphocytic leukemia. PMID- 10364545 TI - The organization of replication and transcription. AB - Models for replication and transcription often display polymerases that track like locomotives along their DNA templates. However, recent evidence supports an alternative model in which DNA and RNA polymerases are immobilized by attachment to larger structures, where they reel in their templates and extrude newly made nucleic acids. These polymerases do not act independently; they are concentrated in discrete "factories," where they work together on many different templates. Evidence for models involving tracking and immobile polymerases is reviewed. PMID- 10364546 TI - Dosage compensation proteins targeted to X chromosomes by a determinant of hermaphrodite fate. AB - In many organisms, master control genes coordinately regulate sex-specific aspects of development. SDC-2 was shown to induce hermaphrodite sexual differentiation and activate X chromosome dosage compensation in Caenorhabditis elegans. To control these distinct processes, SDC-2 acts as a strong gene specific repressor and a weaker chromosome-wide repressor. To initiate hermaphrodite development, SDC-2 associates with the promoter of the male sex determining gene her-1 to repress its transcription. To activate dosage compensation, SDC-2 triggers assembly of a specialized protein complex exclusively on hermaphrodite X chromosomes to reduce gene expression by half. SDC 2 can localize to X chromosomes without other components of the dosage compensation complex, suggesting that SDC-2 targets dosage compensation machinery to X chromosomes. PMID- 10364547 TI - Importance of AMPA receptors for hippocampal synaptic plasticity but not for spatial learning. AB - Gene-targeted mice lacking the L-alpha-amino-3-hydroxy-5-methylisoxazole-4 propionate (AMPA) receptor subunit GluR-A exhibited normal development, life expectancy, and fine structure of neuronal dendrites and synapses. In hippocampal CA1 pyramidal neurons, GluR-A-/- mice showed a reduction in functional AMPA receptors, with the remaining receptors preferentially targeted to synapses. Thus, the CA1 soma-patch currents were strongly reduced, but glutamatergic synaptic currents were unaltered; and evoked dendritic and spinous Ca2+ transients, Ca2+-dependent gene activation, and hippocampal field potentials were as in the wild type. In adult GluR-A-/- mice, associative long-term potentiation (LTP) was absent in CA3 to CA1 synapses, but spatial learning in the water maze was not impaired. The results suggest that CA1 hippocampal LTP is controlled by the number or subunit composition of AMPA receptors and show a dichotomy between LTP in CA1 and acquisition of spatial memory. PMID- 10364549 TI - Reconstructing phylogeny with and without temporal data. AB - Conventional cladistic methods of inferring evolutionary relationships exclude temporal data from the initial search for optimal hypotheses, but stratocladistics includes such data. A comparison of the ability of these methods to recover known, simulated evolutionary histories given the same, evolved character data shows that stratocladistics recovers the true phylogeny in over twice as many cases as cladistics (42 versus 18 percent). The comparison involved 550 unique taxon-by-character matrices, representing 15 evolutionary models and fossil records ranging from 100 to 10 percent complete. PMID- 10364548 TI - Rapid spine delivery and redistribution of AMPA receptors after synaptic NMDA receptor activation. AB - To monitor changes in alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptor distribution in living neurons, the AMPA receptor subunit GluR1 was tagged with green fluorescent protein (GFP). This protein (GluR1-GFP) was functional and was transiently expressed in hippocampal CA1 neurons. In dendrites visualized with two-photon laser scanning microscopy or electron microscopy, most of the GluR1-GFP was intracellular, mimicking endogenous GluR1 distribution. Tetanic synaptic stimulation induced a rapid delivery of tagged receptors into dendritic spines as well as clusters in dendrites. These postsynaptic trafficking events required synaptic N-methyl-D-aspartate (NMDA) receptor activation and may contribute to the enhanced AMPA receptor-mediatedtransmission observed during long-term potentiation and activity-dependent synaptic maturation. PMID- 10364550 TI - Two-dimensional photonic band-Gap defect mode laser AB - A laser cavity formed from a single defect in a two-dimensional photonic crystal is demonstrated. The optical microcavity consists of a half wavelength-thick waveguide for vertical confinement and a two-dimensional photonic crystal mirror for lateral localization. A defect in the photonic crystal is introduced to trap photons inside a volume of 2.5 cubic half-wavelengths, approximately 0.03 cubic micrometers. The laser is fabricated in the indium gallium arsenic phosphide material system, and optical gain is provided by strained quantum wells designed for a peak emission wavelength of 1.55 micrometers at room temperature. Pulsed lasing action has been observed at a wavelength of 1.5 micrometers from optically pumped devices with a substrate temperature of 143 kelvin. PMID- 10364551 TI - Optical properties of an ionic-type phononic crystal AB - An ionic-type phononic crystal composed of two ferroelectric media with opposite spontaneous polarization aligned periodically in a superlattice structure was studied theoretically and experimentally. The coupling between vibrations of the superlattice and the electromagnetic waves results in various long-wavelength optical properties, such as microwave absorption, dielectric abnormality, and polariton excitation, that exist originally in ionic crystals. The results show that this artificial crystal structure can be used to simulate the microscopic physical processes in real crystals. PMID- 10364552 TI - Middle eocene seawater pH and atmospheric carbon dioxide concentrations AB - The carbon dioxide content of the atmosphere [measured as the partial pressure of CO2 (pCO2)] affects the content of the surface ocean, which in turn affects seawater pH. The boron isotope composition (delta11B) of contemporaneous planktonic foraminifera that calcified their tests at different water depths can be used to reconstruct the pH-depth profile of ancient seawater. Construction of a pH profile for the middle Eocene tropical Pacific Ocean shows that atmospheric pCO2 was probably similar to modern concentrations or slightly higher. PMID- 10364553 TI - Regular and irregular patterns in semiarid vegetation AB - Vegetation in many semiarid regions is strikingly patterned, forming regular stripes on hillsides and irregular mosaics on flat ground. A simple model of plant and water dynamics based on ecologically realistic assumptions and with reasonable parameter values captures both of these types of patterns. The regular patterns result from a Turing-like instability; the irregular patterns arise when the ecological dynamics amplify slight small-scale topographic variability. Because of the close agreement between observations and these theoretical results, this system provides a clear example of how nonlinear mechanisms can be important in determining the spatial structure of plant communities. PMID- 10364554 TI - Reexamining fire suppression impacts on brushland fire regimes AB - California shrubland wildfires are increasingly destructive, and it is widely held that the problem has been intensified by fire suppression, leading to larger, more intense wildfires. However, analysis of the California Statewide Fire History Database shows that, since 1910, fire frequency and area burned have not declined, and fire size has not increased. Fire rotation intervals have declined, and fire season has not changed, implying that fire intensity has not increased. Fire frequency and population density were correlated, and it is suggested that fire suppression plays a critical role in offsetting potential impacts of increased ignitions. Large fires were not dependent on old age classes of fuels, and it is thus unlikely that age class manipulation of fuels can prevent large fires. Expansion of the urban-wildland interface is a key factor in wildland fire destruction. PMID- 10364555 TI - Positive feedbacks in the fire dynamic of closed canopy tropical forests AB - The incidence and importance of fire in the Amazon have increased substantially during the past decade, but the effects of this disturbance force are still poorly understood. The forest fire dynamics in two regions of the eastern Amazon were studied. Accidental fires have affected nearly 50 percent of the remaining forests and have caused more deforestation than has intentional clearing in recent years. Forest fires create positive feedbacks in future fire susceptibility, fuel loading, and fire intensity. Unless current land use and fire use practices are changed, fire has the potential to transform large areas of tropical forest into scrub or savanna. PMID- 10364556 TI - The nature of the principal type 1 interferon-producing cells in human blood. AB - Interferons (IFNs) are the most important cytokines in antiviral immune responses. "Natural IFN-producing cells" (IPCs) in human blood express CD4 and major histocompatibility complex class II proteins, but have not been isolated and further characterized because of their rarity, rapid apoptosis, and lack of lineage markers. Purified IPCs are here shown to be the CD4(+)CD11c- type 2 dendritic cell precursors (pDC2s), which produce 200 to 1000 times more IFN than other blood cells after microbial challenge. pDC2s are thus an effector cell type of the immune system, critical for antiviral and antitumor immune responses. PMID- 10364557 TI - Math1: an essential gene for the generation of inner ear hair cells. AB - The mammalian inner ear contains the cochlea and vestibular organs, which are responsible for hearing and balance, respectively. The epithelia of these sensory organs contain hair cells that function as mechanoreceptors to transduce sound and head motion. The molecular mechanisms underlying hair cell development and differentiation are poorly understood. Math1, a mouse homolog of the Drosophila proneural gene atonal, is expressed in inner ear sensory epithelia. Embryonic Math1-null mice failed to generate cochlear and vestibular hair cells. This gene is thus required for the genesis of hair cells. PMID- 10364558 TI - Crystal structure of the Zalpha domain of the human editing enzyme ADAR1 bound to left-handed Z-DNA. AB - The editing enzyme double-stranded RNA adenosine deaminase includes a DNA binding domain, Zalpha, which is specific for left-handed Z-DNA. The 2.1 angstrom crystal structure of Zalpha complexed to DNA reveals that the substrate is in the left handed Z conformation. The contacts between Zalpha and Z-DNA are made primarily with the "zigzag" sugar-phosphate backbone, which provides a basis for the specificity for the Z conformation. A single base contact is observed to guanine in the syn conformation, characteristic of Z-DNA. Intriguingly, the helix-turn helix motif, frequently used to recognize B-DNA, is used by Zalpha to contact Z DNA. PMID- 10364559 TI - Specific coupling of NMDA receptor activation to nitric oxide neurotoxicity by PSD-95 protein. AB - The efficiency with which N-methyl-D-aspartate receptors (NMDARs) trigger intracellular signaling pathways governs neuronal plasticity, development, senescence, and disease. In cultured cortical neurons, suppressing the expression of the NMDAR scaffolding protein PSD-95 (postsynaptic density-95) selectively attenuated excitotoxicity triggered via NMDARs, but not by other glutamate or calcium ion (Ca2+) channels. NMDAR function was unaffected, because receptor expression, NMDA currents, and 45Ca2+ loading were unchanged. Suppressing PSD-95 blocked Ca2+-activated nitric oxide production by NMDARs selectively, without affecting neuronal nitric oxide synthase expression or function. Thus, PSD-95 is required for efficient coupling of NMDAR activity to nitric oxide toxicity, and imparts specificity to excitotoxic Ca2+ signaling. PMID- 10364560 TI - Direct demonstration of P-selectin- and VCAM-1-dependent mononuclear cell rolling in early atherosclerotic lesions of apolipoprotein E-deficient mice. AB - Apolipoprotein E-deficient (ApoE-/-) mice develop atherosclerotic lesions throughout the arterial tree, including the carotid bifurcation. Although the expression of adhesion molecules such as ICAM-1, vascular cell adhesion molecule 1 (VCAM-1), and P-selectin on endothelium that overlie atherosclerotic plaques has been implicated in monocyte recruitment to developing lesions, monocyte adhesion in atherosclerotic vessels has not been observed directly. To investigate which adhesion molecules may be important in monocyte adhesion to atherosclerotic lesions, an isolated mouse carotid artery preparation was developed and perfused with mononuclear cells. We show rolling and attachment of the human monocytic cell line U937 and the mouse monocyte-macrophage cell line P388D1 in carotid arteries from 10- to 12-week-old ApoE-/- and C57BL/6 wild-type mice fed a Western-type diet (21% fat wt/wt) for 4 to 5 weeks. No rolling was observed in carotid arteries from C57BL/6 or BALB/c wild-type mice fed a chow diet and little was observed in BALB/c mice fed a Western-type diet. This model represents early lesion development as shown by minimal macrophage infiltration in the intima of carotid arteries from ApoE-/- mice fed a Western-type diet. Rolling was observed at shear stresses that were characteristic of the low-shear recirculation zone near the carotid bifurcation. Mononuclear cell attachment and rolling were significantly inhibited by monoclonal antibody blockade of P selectin or its leukocyte ligand P-selectin glycoprotein ligand-1. Rolling velocities increased after monoclonal antibody blockade of mononuclear cell alpha4-integrin or VCAM-1, which indicates that alpha4-integrin interacting with VCAM-1 stabilizes rolling interactions and prolongs monocyte transit times. PMID- 10364561 TI - Emigrated neutrophils regulate ventricular contractility via alpha4 integrin. AB - We have previously shown that CD18 and alpha4 integrin were important in the adherence of emigrated neutrophils to cardiac myocytes. Whether either of these molecules is important in myocyte dysfunction is unclear. In this study, we measured contractility as an index of myocyte function. Control contractility was compared with shortening response in myocytes exposed to neutrophils in the presence and absence of anti-CD18 or anti-alpha4 antibodies. Control unloaded cell shortening, expressed as a percentage of resting cell length, measured 10.06+/-1.16% (n=10) at 5 minutes. Circulating neutrophils caused a 35% reduction in cell shortening, an event prevented by anti-CD18, but not by anti-alpha4 antibody. When emigrated neutrophils were added to the myocytes, a profound reduction (50%) in unloaded cell shortening was noted. A significant increase in CD18 and alpha4 integrin was found on emigrated neutrophils. Addition of anti CD18 antibody did not protect the myocyte from the emigrated neutrophils, whereas the addition of an anti-alpha4 antibody significantly reduced neutrophil-induced cell shortening, despite some neutrophils still adhering to the myocytes. Furthermore, emigrated neutrophils were able to cause myocytes to go into contracture within 5 minutes in the presence of neutrophils with or without anti CD18 antibody. In addition to the impairment in unloaded cell shortening, at later times (10 minutes), neutrophils also caused a 40% reduction in the rate of contraction and relaxation. The addition of either anti-CD18 or anti-alpha4 antibody protected the myocytes from these changes. The data suggest that immunosuppression of CD18 on emigrated neutrophils was only partially effective in reducing myocyte dysfunction. In contrast, immunosuppression of the alpha4 integrin alone was sufficient to dramatically reduce all parameters of cell dysfunction measured in this study. PMID- 10364562 TI - Strain differences in neointimal hyperplasia in the rat. AB - We performed an initial screen of 11 rat strains by use of a standard balloon injury to the left iliac artery to observe whether genetically determined differences existed in the development of neointimal hyperplasia. Neointimal hyperplasia was assayed 8 weeks after the vascular injury on coded microscopic sections. Statistically significant differences in the percentages of the vascular wall cross-sectional areas composed of intima (percentage intima) secondary to neointimal hyperplasia were noted among the different rat strains (P<0.02), with the Brown-Norway (BN), Dark Agouti, and Milan normotensive strain rats having the highest and the spontaneously hypertensive rats (SHR) having the lowest percentages of intima. In a separate experiment, F1 hybrids of SHRxBN strains and parental BN and SHR underwent the vascular injury, and the parental strains again showed a statistically significant difference from one another in the mean percentage of intima (P<0. 0001). The F1 hybrids showed an average percentage of intima intermediate between those of the parental strains. The average lumen size of the injured BN vessels were significantly smaller than that of the noninjured control vessels (P=0.044), but this significance disappeared when the circular areas of these vessels were calculated without taking neointimal growth into consideration (P=0.649). These results provide the groundwork for a genetic linkage analysis to identify the genes that influence the development of neointimal hyperplasia after vascular injury. PMID- 10364564 TI - Vitaxin, a humanized monoclonal antibody to the vitronectin receptor (alphavbeta3), reduces neointimal hyperplasia and total vessel area after balloon injury in hypercholesterolemic rabbits. AB - The vitronectin receptor (alphavbeta3) mediates several biological processes that are critical to the formation of a neointima after coronary interventions. Blockade of alphavbeta3 could reduce neointima formation by inhibiting smooth muscle cell migration, decreasing transforming growth factor-beta1 expression, enhancing apoptosis, or reducing neovasculature. The effects of short-term administration of Vitaxin, a humanized monoclonal antibody to alphavbeta3, on the responses to balloon injury were tested in hyperlipidemic rabbits. Balloon angioplasty was performed on the iliac arteries of male New Zealand White rabbits that were fed an atherogenic diet for 1 week before injury and until euthanization at 4 weeks. Rabbits were given either saline (control) or 1 of 2 dosing regimens of Vitaxin (high dose, 5.0 mg/kg, and low dose, 0.5 mg/kg), which were administered intra-arterially before injury and intramuscularly on days 2 and 3. High-dose and low-dose Vitaxin were equally effective in decreasing neointima formation even in the presence of hypercholesterolemia, a stimulus to alphavbeta3 expression. Vitaxin reduced transforming growth factor-beta1 and enhanced apoptosis in injured arteries. Despite these positive effects, Vitaxin administration was associated with a reduction in artery size, indicating a negative effect on remodeling. Vitaxin has a potential role in preventing intimal hyperplasia, especially if the negative effects on remodeling can be overcome, by dose adjustment or other strategies. PMID- 10364563 TI - GC factor 2 represses platelet-derived growth factor A-chain gene transcription and is itself induced by arterial injury. AB - Platelet-derived growth factor (PDGF) is a mitogen and chemoattractant for a wide variety of cell types. The genes encoding PDGF A chain (PDGF-A) and PDGF B chain (PDGF-B) reside on separate chromosomes and are independently regulated at the level of transcription. Regulatory events underlying inducible PDGF-A expression have been the focus of much investigation. However, mechanisms that inhibit transcription of this gene are not well understood. In this study, we report the capacity of a newly cloned DNA binding factor, GC factor 2 (GCF2), to repress expression driven by the human PDGF-A promoter. 5' Deletion and transient cotransfection analysis in vascular endothelial cells revealed that GCF2 repression is mediated by a nucleotide region located in the proximal region of the PDGF-A promoter. Electrophoretic mobility shift assays demonstrate that GCF2 binds to this region in a specific and dose-dependent manner. Interestingly, the site bound by GCF2 overlaps those for specificity protein-1 (Sp1) and early growth response factor-1 (Egr-1), zinc finger transcription factors that direct basal and inducible expression of the PDGF-A gene. Gel shift experiments revealed that GCF2 competes with these factors for interaction with the PDGF-A promoter. Overexpression of GCF2 suppressed endogenous PDGF-A expression in vascular endothelial cells and smooth muscle cells. GCF2 was induced on mechanical injury of cells in culture as well as after balloon injury of the rat carotid artery wall. Time course studies revealed the sustained induction of GCF2 after injury while PDGF-A levels sharply returned to baseline. Smooth muscle cell proliferation was inhibited by GCF2, an effect reversed by the addition of exogenous PDGF-AA. These findings demonstrate negative regulation of PDGF-A expression by GCF2. This is the first report of the induction of an endogenous transcriptional repressor in the rat vessel wall. PMID- 10364565 TI - Multiple connexins form gap junction channels in rat basilar artery smooth muscle cells. AB - Three connexins, Cx43, Cx40, and Cx37, have been found by protein or mRNA analysis to be prominent in mammalian blood vessels, but electrophysiological characterization of gap junction channels in freshly isolated vascular smooth muscle cells (SMCs) has not previously been reported. We used a dual-perforated patch-clamp technique to study gap junction conductances in SMC pairs from rat basilar arteries. Macroscopic junctional conductance (Gj) measured in 98 cell pairs with either Cs+ or K+ ranged between 0.68 and 24.8 nS. In weakly coupled cells (Gj<5 nS), single-channel currents were readily resolved without pharmacological uncoupling agents, allowing identification of 4 major unitary conductances. Two of these conductances, 80 to 120 pS and 150 to 200 pS, corresponded to the major conductance states for homotypic channels formed from Cx43 or Cx40, which we confirmed were present in smooth muscle by immunofluorescence analysis. Two other conductances, 220 to 280 pS and >300 pS, were identified that have not been previously reported in vascular SMCs. Macroscopic recordings revealed currents that deactivated incompletely over a broad range of transjunctional potentials. In about half of the pairs, we identified macroscopic as well as single-channel currents that exhibited marked voltage asymmetry, consistent with nonhomotypic, ie, either heterotypic or heteromeric channels. Our data indicate that basilar artery SMCs are coupled in vivo in a richly complex manner, involving Cx43, Cx40, and other large conductance channels, and that a significant number of couplings involve putative nonhomotypic channels. PMID- 10364566 TI - Localized expression of aromatase in human vascular tissues. AB - The atheroprotective effects of estrogen are well established and the presence of an estrogen receptor in vascular tissues has recently been reported. Therefore, we investigated the localization of the estrogen-producing enzyme aromatase in vascular tissues to assess the possible contribution of endocrine, paracrine, and autocrine modes of action. Aromatase was found in human vascular smooth muscle cells (SMCs) but not in endothelial cells on in situ hybridization. These observations were further supported by quantitative analysis of aromatase mRNA and the activity in 15 human vascular specimens. Only trace levels of expression were detected in the 3 infants examined, whereas 0.0088 to 0.0806 amol/ microg RNA of aromatase mRNA and 12.9 to 122.3 fmol. h-1. mg-1 protein of the activity were detected in 12 of the adult individuals. The switching of tissue-specific exon 1 of the human aromatase gene was also observed in some cases. Aromatase was found to be expressed only in cultured SMCs and not in cultured endothelial cells of human aorta and pulmonary artery and to be regulated through dexamethasone and the signaling pathways of protein kinase A and C. Study results revealed the localized expression of aromatase in vascular SMCs, which indicated a possible direct action of locally produced estrogen in an autocrine or paracrine manner, with possible cross talk between smooth muscle and endothelial cells. PMID- 10364567 TI - Structural plasticity of the cardiac nuclear pore complex in response to regulators of nuclear import. AB - Communication between the cytoplasm and nucleoplasm of cardiac cells occurs by molecular transport through nuclear pores. In lower eukaryotes, nuclear transport requires the maintenance of cellular energetics and ion homeostasis. Although heart muscle is particularly sensitive to metabolic stress, the regulation of nuclear transport through nuclear pores in cardiomyocytes has not yet been characterized. With the use of laser confocal and atomic force microscopy, we observed nuclear transport in cardiomyocytes and the structure of individual nuclear pores under different cellular conditions. In response to the depletion of Ca2+ stores or ATP/GTP pools, the cardiac nuclear pore complex adopted 2 distinct conformations that led to different patterns of nuclear import regulation. Depletion of Ca2+ indiscriminately prevented the nuclear import of macromolecules through closure of the nuclear pore opening. Depletion of ATP/GTP only blocked facilitated transport through a simultaneous closure of the pore and relaxation of the entire complex, which allowed other molecules to pass into the nucleus through peripheral routes. The current study of the structural plasticity of the cardiac nuclear pore complex, which was observed in response to changes in cellular conditions, identifies a gating mechanism for molecular translocation across the nuclear envelope of cardiac cells. The cardiac nuclear pore complex serves as a conduit that differentially regulates nuclear transport of macromolecules and provides a mechanism for the control of nucleocytoplasmic communication in cardiac cells, in particular under stress conditions associated with disturbances in cellular bioenergetics and Ca2+ homeostasis. PMID- 10364568 TI - Modulation of iron uptake in heart by L-type Ca2+ channel modifiers: possible implications in iron overload. AB - Heart failure is the leading cause of mortality in patients with transfusional iron (Fe) overload in which myocardial iron uptake ensues via a transferrin independent process. We examined the ability of L-type Ca2+ channel modifiers to alter Fe2+ uptake by isolated rat hearts and ventricular myocytes. Perfusion of rat hearts with 100 nmol/L 59Fe2+ and 5 mmol/L ascorbate resulted in specific 59Fe2+ uptake of 20.4+/-1.9 ng of Fe per gram dry wt. Abolishing myocardial electrical excitability with 20 mmol/L KCl reduced specific 59Fe2+ uptake by 60+/ 7% (P<0.01), which suggested that a component of myocardial Fe2+ uptake depends on membrane voltage. Accordingly, 59Fe2+ uptake was inhibited by 10 micromol/L nifedipine (45+/-12%, P<0.02) and 100 micromol/L Cd2+ (86+/-3%; P<0. 001) while being augmented by 100 nmol/L Bay K 8644 (61+/-18%, P<0. 01) or 100 nmol/L isoproterenol (40+/-12%, P<0.05). By contrast, uptake of 100 nmol/L ferric iron (59Fe3+) was significantly lower (1. 4+/-0.3 ng Fe per gram dry wt; P<0.001) compared with divalent iron. These data suggest that a component of Fe2+ uptake into heart occurs via the L-type Ca2+ channel in myocytes. To investigate this further, the effects of Fe2+ on cardiac myocyte L-type Ca2+ currents were measured. In the absence of Ca2+, noninactivating nitrendipine-sensitive Fe2+ currents were recorded with 15 mmol/L [Fe2+]o. Low concentrations of Fe2+ enhanced Ca2+ current amplitude and slowed inactivation rates, which was consistent with Fe2+ entry into the cell, whereas higher Fe2+ levels caused dose dependent decreases in peak current. Fe3+ had no effect on current amplitude or decay. Combined, our data suggest that myocardial Fe2+ uptake occurs via L-type Ca2+ channels and that blockade of these channels might be useful in the treatment of patients with excessive serum iron levels. PMID- 10364569 TI - Effects of myosin heavy chain isoform switching on Ca2+-activated tension development in single adult cardiac myocytes. AB - Cardiac myosin heavy chain (MHC) isoforms are known to play a key role in defining the dynamic contractile behavior of the heart during development. It remains unclear, however, whether cardiac MHC isoforms influence other important features of cardiac contractility, including the Ca2+ sensitivity of isometric tension development. To address this question, adult rats were treated chemically to induce the hypothyroid state and cause a transition in the ventricular cardiac MHC isoform expression pattern from predominantly the alpha-MHC isoform to exclusively the beta-MHC isoform. We found a significant desensitization in the Ca2+ sensitivity of tension development in beta-MHC-expressing ventricular myocytes (pCa50=5. 51+/-0.03, where pCa is -log[Ca2+], and pCa50 is pCa at which tension is one-half maximal) compared with that in predominantly alpha-MHC expressing myocytes (pCa50=5.68+/-0.05). No differences between the 2 groups were observed in the steepness of the tension-pCa relationship or in the maximum isometric force generated. Instantaneous stiffness measurements were made that provide a relative measure of changes in the numbers of myosin crossbridges attached to actin during Ca2+ activation. Results showed that the relative stiffness-pCa relationship was shifted to the right in beta-MHC-expressing myocytes compared with the alpha-MHC-expressing cardiac myocytes (pCa50=5.47+/ 0.05 versus 5.76+/-0.05, respectively). We conclude that MHC isoform switching in adult cardiac myocytes alters the Ca2+ sensitivity of stiffness and tension development. These results suggest that the activation properties of the thin filament are in part MHC isoform dependent in cardiac muscle, indicating an additional role for MHC isoforms in defining cardiac contractile function. PMID- 10364570 TI - Spatiotemporal heterogeneity in the induction of ventricular fibrillation by rapid pacing: importance of cardiac restitution properties. AB - The mechanism by which rapid pacing induces ventricular fibrillation (VF) is unclear. We performed computerized epicardial mapping studies in 10 dogs, using 19-beat pacing trains. The pacing interval (PI) of the first train was 300 ms and then was progressively shortened until VF was induced. For each PI, we constructed restitution curves for the effective refractory period (ERP). When the PI was long, the activation cycle length (CL) was constant throughout the mapped region. However, as the PI shortened, there was an increase in the spatiotemporal complexity of the CL variations and an increase in the slope of the ERP restitution curve. In 5 dogs, we documented the initiation of VF by wavebreak at the site of long-short CL variations. Computer simulation studies using the Luo-Rudy I ventricular action potential model in simulated 2 dimensional tissue reproduced the experimental results when normal ERP and conduction velocity (CV) restitution properties were intact. By altering CV and ERP restitutions in this model, we found that CV restitution creates spatial CL variations, whereas ERP restitution underlies temporal, beat-to-beat variations in refractoriness during rapid pacing. Together, the interaction of CV and ERP restitutions produces spatiotemporal oscillations in cardiac activation that increase in amplitude as the PI decreases, ultimately causing wavebreak at the site of intrinsic heterogeneity. This initial wavebreak then leads to the formation of spiral waves and VF. These findings support a key role for both CV and ERP restitutions in the initiation of VF by rapid pacing. PMID- 10364571 TI - The angiotensin II-dependent association of Jak2 and c-Src requires the N terminus of Jak2 and the SH2 domain of c-Src. AB - The binding of angiotensin II (Ang II) to AT1 is known to increase the kinase activity of several nonreceptor tyrosine kinases including Jak2 and c-Src. In the present study, we demonstrate that treatment of vascular smooth muscle cells with Ang II results in a rapid and transient association of Jak2 and c-Src. This association is dependent on a catalytically active Jak2 kinase, because it is blocked both by pharmacological means and by the inability of a catalytically inactive Jak2 to associate with c-Src. c-Src bound tyrosine phosphorylated Jak2 but was unable to bind an equal amount of unphosphorylated Jak2 protein, indicating that the SH2 domain of c-Src mediates this association. In vivo studies indicated that c-Src binds the N-terminus of Jak2 as expression of a Jak2 molecule lacking the initial 240 amino acids, including 16 tyrosines, and was unable to bind c-Src. Lastly, using transiently transfected COS-7 cells, we found that Ang II treatment induced an association between c-Src and wild-type Jak2 but not between c-Src and the Jak2 molecule that lacks the initial 240 amino acids. Thus, our data suggest that in addition to increasing the kinase activities Jak2 and c-Src, treatment of cells with Ang II results in the physical association of Jak2 and c-Src; an association that is mediated by the SH2 domain of c-Src and the N-terminus of Jak2. PMID- 10364572 TI - Mechanically driven contour-length adjustment in rat cardiac titin's unique N2B sequence: titin is an adjustable spring. AB - The giant elastic protein titin is largely responsible for passive forces in cardiac myocytes. A number of different titin isoforms with distinctly different structural elements within their central I-band region are expressed in human myocardium. Their coexpression has so far prevented an understanding of the respective contributions of the isoforms to myocardial elasticity. Using isoform specific antibodies, we find in the present study that rat myocardium expresses predominantly the small N2B titin isoform, which allows us to characterize the elastic behavior of this isoform. The extensibility and force response of N2B titin were studied by using immunoelectron microscopy and by measuring the passive force-sarcomere length (SL) relation of single rat cardiac myocytes under a variety of mechanical conditions. Experimental results were compared with the predictions of a mechanical model in which the elastic titin segment behaves as two wormlike chains, the tandem immunoglobulin (Ig) segments and the PEVK segment (rich in proline [P], glutamate [E], valine [V], and lysine [K] residues), connected in series. The overall contour length was predicted from the sequence of N2B cardiac titin. According to mechanical measurements, above approximately 2.2 microm SL titin's elastic segment extends beyond its predicted contour length. Immunoelectron microscopy indicates that a prominent source of this contour-length gain is the extension of the unique N2B sequence (located between proximal tandem Ig segment and PEVK), and that Ig domain unfolding is negligible. Thus, the elastic region of N2B cardiac titin consists of three mechanically distinct extensible segments connected in series: the tandem Ig segment, the PEVK segment, and the unique N2B sequence. Rate-dependent and repetitive stretch release experiments indicate that both the contour-length gain and the recovery from it involve kinetic processes, probably unfolding and refolding within the N2B segment. As a result, the contour length of titin's extensible segment depends on the rate and magnitude of the preceding mechanical perturbations. The rate of recovery from the length gain is slow, ensuring that the adjusted length is maintained through consecutive cardiac cycles and that hysteresis is minimal. Thus, as a result of the extensible properties of the unique N2B sequence, the I band region of the N2B cardiac titin isoform functions as a molecular spring that is adjustable. PMID- 10364573 TI - Monocyte rolling in early atherogenesis: vital role in lesion development. PMID- 10364574 TI - Molecular diagnosis of herpes simplex virus infections in the central nervous system. PMID- 10364575 TI - Borrelia burgdorferi in tick cell culture modulates expression of outer surface proteins A and C in response to temperature. AB - The Lyme disease spirochete Borrelia burgdorferi sensu stricto downregulates outer surface protein A (OspA) and upregulates outer surface protein C (OspC) during tick feeding. The switching of these proteins correlates with increased spirochetal infectivity for the mammal. We examined the effect of temperature on differential expression of OspA and OspC by B. burgdorferi cocultivated with a cell line isolated from the vector tick Ixodes scapularis. The effect of incubation at 31, 34, or 37 degrees C on expression of OspA and OspC by B. burgdorferi JMNT and N40 was analyzed by indirect fluorescent-antibody microscopy, polyacrylamide gel electrophoresis, and immunoblotting. The amount of OspA relative to the amount of flagellin was highest in spirochetes cocultivated with tick cells at 31 degrees C and declined with increasing temperature in both strains. OspC production was enhanced in spirochetes cocultivated with tick cells at 37 degrees C. Spirochetes grown axenically in BSK-H medium also produced more OspC at 37 degrees C, but OspA content was not appreciably affected by temperature. Our findings indicate that temperature, along with cultivation in a tick cell culture system, plays a role in the differential expression of OspA and enhances differential expression of OspC by spirochetes. PMID- 10364576 TI - Subtyping of Haemophilus influenzae strains by pulsed-field gel electrophoresis. AB - A total of 200 isolates of Haemophilus influenzae were analyzed by serotyping, biotyping, and pulsed-field gel electrophoresis (PFGE). A total of 178 epidemiologically unrelated strains of H. influenzae demonstrated a variety of genome patterns by PFGE, and 165 genotypes were thus obtained in this study. PFGE typing proved to have a much stronger discriminatory power than either serotyping or biotyping. Six serotype b strains were all classified into discrete genotypes. A PFGE analysis of 18 strains obtained from the nasopharynx, blood, and cerebrospinal fluid of patients with meningitis also supported the hypothesis that invasive H. influenzae disseminates from the nasopharynx to the bloodstream and then subsequently to other body sites. PFGE typing of 10 other strains isolated from household contacts of patients with H. influenzae infection revealed that the strain that caused the H. influenzae infection often colonized the nasopharynges of household contacts. Our findings suggest that PFGE analysis is useful for the epidemiological study of H. influenzae infection, even when the invasive disease is caused by serotype b strains. PMID- 10364577 TI - Proficiency of clinical laboratories in Spain in detecting vancomycin-resistant Enterococcus spp. The Spanish VRE Study Group. AB - Studies in a variety of U.S. clinical laboratories have demonstrated difficulty in detecting intermediate and low-level vancomycin-resistant enterococci (VRE). The misclassification of "at least intermediate resistant isolates" as vancomycin susceptible may have both clinical implications and a negative impact on measures to control the spread of VRE. No published study has assessed the ability of clinical laboratories in Europe to detect VRE. So, the apparent low prevalence of VRE in European hospitals may be, in part, secondary to the inability of these laboratories to detect all VRE. In an effort to assess European laboratories' proficiency in detecting VRE, we identified 22 laboratories in Spain and asked them to test four VRE strains and one susceptible enterococcal strain from the Centers for Disease Control and Prevention collection. Each organism was tested by the routine antimicrobial susceptibility testing method used by each laboratory. Overall, VRE were correctly identified in 61 of 88 (69.1%) instances. The accuracy of VRE detection varied with the level of resistance and the antimicrobial susceptibility method. The high-level-resistant strain (Enterococcus faecium; MIC, 512 microg/ml) was accurately detected in 20 of 22 (91. 3%) instances, whereas the intermediate-resistant isolate (Enterococcus gallinarum; MIC, 8 microg/ml) was accurately detected in only 11 of 22 (50%) instances. Classification errors occurred in 27 of 88 (30.9%) instances. Misclassification as vancomycin susceptible was the most common error (16 of 27 [59.3%] instances). Our study shows that the participating Spanish laboratories had an overall acceptable proficiency in detecting VRE but that a substantial proportion of VRE isolates with low or intermediate levels of resistance were not detected. We recommend that studies be conducted to validate laboratory proficiency testing as an important step in the prevention and control of the spread of antimicrobial resistance. PMID- 10364578 TI - Improved methods for immunoassay of mycothiol. AB - Improved enzyme-linked immunosorbent assay (ELISA) methods have been developed for the determination of femtomole amounts of mycothiol (MSH), the main low molecular-weight thiol in mycobacteria. The immunoassays utilize an affinity purified rabbit polyclonal antibody that is highly specific for the pseudodisaccharide moiety of MSH. MSH was first biotinylated by the thiol specific reagent 3-(N-maleimidopropionyl)biocytin. The MSH-biotin adduct was then captured with immobilized avidin and detected with anti-MSH antibody (biotin capture ELISA) or was captured with immobilized anti-MSH antibody and detected with alkaline phosphatase-labelled avidin (MSH-capture ELISA). The MSH-capture ELISA was the most sensitive method, measuring as little as 0.3 fmol of MSH. Methods for biotinylating MSH directly from Mycobacterium spp. are described. The MSH-capture ELISA was tested for the detection of M. avium seeded in human urine or cerebrospinal fluid samples and for screening mutant M. smegmatis strains to detect MSH production. PMID- 10364579 TI - Identification of Burkholderia spp. in the clinical microbiology laboratory: comparison of conventional and molecular methods. AB - Cystic fibrosis (CF) predisposes patients to bacterial colonization and infection of the lower airways. Several species belonging to the genus Burkholderia are potential CF-related pathogens, but microbiological identification may be complicated. This situation is not in the least due to the poorly defined taxonomic status of these bacteria, and further validation of the available diagnostic assays is required. A total of 114 geographically diverse bacterial isolates, previously identified in reference laboratories as Burkholderia cepacia (n = 51), B. gladioli (n = 14), Ralstonia pickettii (n = 6), B. multivorans (n = 2), Stenotrophomonas maltophilia (n = 3), and Pseudomonas aeruginosa (n = 11), were collected from environmental, clinical, and reference sources. In addition, 27 clinical isolates putatively identified as Burkholderia spp. were recovered from the sputum of Dutch CF patients. All isolates were used to evaluate the accuracy of two selective growth media, four systems for biochemical identification (API 20NE, Vitek GNI, Vitek NFC, and MicroScan), and three different PCR-based assays. The PCR assays amplify different parts of the ribosomal DNA operon, either alone or in combination with cleavage by various restriction enzymes (PCR-restriction fragment length polymorphism [RFLP] analysis). The best system for the biochemical identification of B. cepacia appeared to be the API 20NE test. None of the biochemical assays successfully grouped the B. gladioli strains. The PCR-RFLP method appeared to be the optimal method for accurate nucleic acid-mediated identification of the different Burkholderia spp. With this method, B. gladioli was also reliably classified in a separate group. For the laboratory diagnosis of B. cepacia, we recommend parallel cultures on blood agar medium and selective agar plates. Further identification of colonies with a Burkholderia phenotype should be performed with the API 20NE test. For final confirmation of species identities, PCR amplification of the small-subunit rRNA gene followed by RFLP analysis with various enzymes is recommended. PMID- 10364580 TI - Most Enterobacter aerogenes strains in France belong to a prevalent clone. AB - The aim of this study was to determine the distribution in France of the Enterobacter aerogenes prevalent clone isolated in the hospitals of the Marseille area (A. Davin-Regli, D. Monnet, P. Saux, C. Bosi, R. Charrel, A. Barthelemy, and C. Bollet, J. Clin. Microbiol. 34:1474-1480, 1996). A total of 123 E. aerogenes isolates were collected from 23 hospital laboratories and analyzed by random amplification of polymorphic DNA and enterobacterial repetitive intergenic consensus-PCR to determine their epidemiological relatedness. Molecular typing revealed that 21 of the 23 laboratories had isolated this prevalent clone harboring the plasmid encoding for extended-spectrum beta-lactamase of the TEM-24 type. Most isolates were susceptible only to imipenem and gentamicin. Their dissemination seems to be clonal and was probably the result of the general use of broad-spectrum cephalosporins and quinolones. Four isolates showed an alteration of their outer membrane proteins, causing decrease of susceptibility to third-generation cephalosporins and imipenem and leading to the critical situation of having no alternative therapeutic. The large dissemination of the E. aerogenes prevalent clone probably results from its good adaptation to the antibiotics administered in France and the hospital environment, particularly in intensive care units. PMID- 10364581 TI - Epidemiological study of an Acinetobacter baumannii outbreak by using a combination of antibiotyping and ribotyping. AB - From June to November 1994 (period 1) and from February to June 1995 (period 2), multiresistant Acinetobacter baumannii strains were isolated in intensive care units and surgical wards of the Amiens Teaching Hospital Center (Amiens, France). Eighteen isolates were obtained from 17 (1%) of 1,706 patients admitted during both of these periods, giving an incidence rate of nosocomial infection per 1,000 patient days of 0.6%. Of 17 infected patients, 9 had pneumonia, 3 had urinary tract infection, 2 had peritonitis, 1 had septicemia, 1 had a catheter infection, and 1 had pneumonia and urinary tract infection. According to typing results, four antibiotic resistance profiles were detected: a, b, c, and d; seven ribotypes were distinguished by both restriction enzymes EcoRI and SalI (A, B, C, D, E, F, and G). By combining antibiotyping and ribotyping, we obtained eight groups of strains (groups I to VIII). Group I contained five strains (strains 4, 5, 7, 8, and 9) which had antibiogram pattern a and ribopattern A and constituted the outbreak strains. The strains of group II (strains 3, 10, 11, 13, and 14) were closely related to outbreak strain A and appeared to be variants of ribotype A (A2 [strain 3]; A4 [strain 10]; A5 [strains 11, 13, and 14]). Groups III, IV, V, VI, VII, and VIII included strains which were epidemiologically unrelated to the strains of group I and were considered nonoutbreak strains. PMID- 10364582 TI - Salmonella enteritidis phage types 1 and 4: pheno- and genotypic epidemiology of recent outbreaks in Finland. AB - In the 1990s, Salmonella enterica subsp. enterica serovar Enteritidis has caused 15 outbreaks in Finland; 12 of them were caused by phage type 1 (PT1) and PT4. Thus far, there has been no clear evidence as to the source of these Salmonella Enteritidis PT1 and PT4 strains, so it was necessary to try to characterize them further. Salmonella Enteritidis PT1 (n = 57) and PT4 (n = 43) isolates from different sources were analyzed by genomic pulsed-field gel electrophoresis (PFGE), plasmid profiling, and antimicrobial resistance testing to investigate the distribution of their subtypes in Finland. It was also hoped that this investigation would help in identifying the sources of the infections, especially the sources of the outbreaks caused by PT1 and PT4 in the 1990s. The results showed that both PFGE and plasmid profiling, but not antimicrobial susceptibility testing, were capable of differentiating isolates of Salmonella Enteritidis PT1 and PT4. By genotypic methods, it was possible to divide both PT1 and PT4 isolates into 12 subtypes. It could also be shown that all PT1 outbreak isolates were identical and, at least with this collection of isolates, that the outbreaks did not originate from the Baltic countries or from Russia, where this phage type predominates. It was also established that the outbreaks caused by PT4 all had different origins. Valuable information for future investigations was gained on the distribution of molecular subtypes of strains that originated from the tourist resorts that are popular among Finns and of strains that were isolated from livestock. PMID- 10364583 TI - Evaluation of the discriminatory power of typing methods for Neisseria gonorrhoeae. AB - A panel of 18 strains of Neisseria gonorrhoeae, known to be temporally and geographically diverse, was used to evaluate a number of typing systems, including conventional auxotyping and serotyping and the molecular methods of arbitrarily primed PCR (AP-PCR), amplified ribosomal-DNA restriction analysis (ARDRA), opa typing, and pulsed-field gel electrophoresis (PFGE). The discriminatory power of the different typing methods were determined with a collection of 87 clinical isolates from commercial sex workers in Indonesia, and Simpson's index of diversity was calculated. Of the two traditional techniques, auxotyping and serotyping, the latter gives the highest discriminatory index (DI) (DI, 0.846). The combination of auxotyping and serotyping yields a high DI (DI, 0. 928). D11344- and D8635-primed PCR showed low DIs of 0.608 and 0.622, respectively, but a combination of the two primers had a DI of 0. 849. The combination of serotyping with D11344-primed or D8635-primed PCR resulted in DIs of 0.936 and 0.937, respectively. ARDRA revealed a low DI of 0.743 alone but a DI of 0.955 in combination with serotyping. PFGE using the restriction enzyme BglII and opa typing produced the highest discrimination (DIs, 0.997 and 0. 996, respectively) for isolates of N. gonorrhoeae. PMID- 10364584 TI - Multiple types of Legionella pneumophila serogroup 6 in a hospital heated-water system associated with sporadic infections. AB - Five sporadic cases of nosocomial Legionnaires' disease were documented from 1989 to 1997 in a hospital in northern Italy. Two of them, which occurred in a 75-year old man suffering from ischemic cardiopathy and in an 8-year-old girl suffering from acute leukemia, had fatal outcomes. Legionella pneumophila serogroup 6 was isolated from both patients and from hot-water samples taken at different sites in the hospital. These facts led us to consider the possibility that a single clone of L. pneumophila serogroup 6 had persisted in the hospital environment for 8 years and had caused sporadic infections. Comparison of clinical and environmental strains by monoclonal subtyping, macrorestriction analysis (MRA), and arbitrarily primed PCR (AP-PCR) showed that the strains were clustered into three different epidemiological types, of which only two types caused infection. An excellent correspondence between the MRA and AP-PCR results was observed, with both techniques having high discriminatory powers. However, it was not possible to differentiate the isolates by means of ribotyping and analysis of rrn operon polymorphism. Environmental strains that antigenically and chromosomally matched the infecting organism were present at the time of infection in hot-water samples taken from the ward where the patients had stayed. Interpretation of the temporal sequence of events on the basis of the typing results for clinical and environmental isolates enabled the identification of the ward where the patients became infected and the modes of transmission of Legionella infection. The long term persistence in the hot-water system of different clones of L. pneumophila serogroup 6 indicates that repeated heat-based control measures were ineffective in eradicating the organism. PMID- 10364585 TI - Fluconazole susceptibilities of bloodstream Candida sp. isolates as determined by National Committee for Clinical Laboratory Standards method M27-A and two other methods. AB - The in vitro activity of fluconazole against 143 Candida spp. obtained from the bloodstreams of 143 hospitalized patients from 1995 to 1997 was studied. Susceptibility tests were carried out by two macrodilution methods, the M27-A and a modified M27-A method (0. 165 M, pH 7/morpholinepropanesulfonic acid-buffered RPMI 1640 medium supplemented with 20 g of D-dextrose per liter), and by the agar diffusion method (with 15-microg fluconazole [Neo-Sensitab] tablets). With 2 microg of fluconazole per ml, 96.92% of 65 C. albicans isolates, 86.2% of 58 C. parapsilosis isolates 7 of 8 C. tropicalis isolates, and 1 of 6 C. glabrata isolates were inhibited. Only one strain of C. albicans and one strain of C. tropicalis were resistant. The agreement between the two macrodilution methods was greater than 90% within +/-2 log2 dilutions for all strains except C. glabrata (83.3%) and C. tropicalis (87.5%). Generally, MICs were 1 log2 dilution lower in glucose-supplemented RPMI 1640 medium. No correlation between zone sizes and MICs was found. All strains susceptible by the diffusion test were susceptible by the dilution method, but the converse was not necessarily true. Interestingly, inhibition zones were smaller for C. albicans, for which the geometric mean MIC was 0.29 microg/ml and the mean inhibition zone diameter was 25.7 mm, while for C. parapsilosis the geometric mean MIC was 0.96 microg/ml and the mean inhibition zone diameter was 31. 52 mm. In conclusion, the two macrodilution methods give similar results. The modified M27-A method with 2% dextrose has the advantage of shortening the incubation time and simplifying the endpoint determination. PMID- 10364586 TI - Differentiation of Burkholderia species by PCR-restriction fragment length polymorphism analysis of the 16S rRNA gene and application to cystic fibrosis isolates. AB - Burkholderia cepacia, which is an important pathogen in cystic fibrosis (CF) owing to the potential severity of the infections and the high transmissibility of some clones, has been recently shown to be a complex of five genomic groups, i.e., genomovars I, II (B. multivorans), III, and IV and B. vietnamiensis. B. gladioli is also involved, though rarely, in CF. Since standard laboratory procedures fail to provide an accurate identification of these organisms, we assessed the ability of restriction fragment length polymorphism (RFLP) analysis of amplified 16S ribosomal DNA (rDNA), with the combination of the patterns obtained with six endonucleases, to differentiate Burkholderia species. This method was applied to 16 type and reference strains of the genus Burkholderia and to 51 presumed B. cepacia clinical isolates, each representative of one clone previously determined by PCR ribotyping. The 12 Burkholderia type strains tested were differentiated, including B. cepacia, B. multivorans, B. vietnamiensis, and B. gladioli, but neither the genomovar I and III reference strains nor the genomovar IV reference strain and B. pyrrociniaT were distinguishable. CF clinical isolates were mainly distributed in RFLP group 2 (which includes B. multivoransT) and RFLP group 1 (which includes B. cepacia genomovar I and III reference strains, as well as nosocomial clinical isolates). Two of the five highly transmissible clones in French CF centers belonged to RFLP group 2, and three belonged to RFLP group 1. The remaining isolates either clustered with other Burkholderia species (B. cepacia genomovar IV or B. pyrrocinia, B. vietnamiensis, and B. gladioli) or harbored unique combinations of patterns. Thus, if further validated by hybridization studies, PCR-RFLP of 16S rDNA could be an interesting identification tool and contribute to a better evaluation of the respective clinical risks associated with each Burkholderia species or genomovar in patients with CF. PMID- 10364587 TI - Molecular subtyping of Clostridium perfringens by pulsed-field gel electrophoresis to facilitate food-borne-disease outbreak investigations. AB - Clostridium perfringens is a common cause of food-borne illness. The illness is characterized by profuse diarrhea and acute abdominal pain. Since the illness is usually self-limiting, many cases are undiagnosed and/or not reported. Investigations are often pursued after an outbreak involving large numbers of people in institutions, at restaurants, or at catered meals. Serotyping has been used in the past to assist epidemiologic investigations of C. perfringens outbreaks. However, serotyping reagents are not widely available, and many isolates are often untypeable with existing reagents. We developed a pulsed-field gel electrophoresis (PFGE) method for molecular subtyping of C. perfringens isolates to aid in epidemiologic investigations of food-borne outbreaks. Six restriction endonucleases (SmaI, ApaI, FspI, MluI, KspI, and XbaI) were evaluated with a select panel of C. perfringens strains. SmaI was chosen for further studies because it produced 11 to 13 well-distributed bands of 40 to approximately 1,100 kb which provided good discrimination between isolates. Seventeen distinct patterns were obtained with 62 isolates from seven outbreak investigations or control strains. In general, multiple isolates from a single individual had indistinguishable PFGE patterns. Epidemiologically unrelated isolates (outbreak or control strains) had unique patterns; isolates from different individuals within an outbreak had similar, if not identical, patterns. PFGE identifies clonal relationships of isolates which will assist epidemiologic investigations of food-borne-disease outbreaks caused by C. perfringens. PMID- 10364588 TI - Detection and identification of Ehrlichia, Borrelia burgdorferi sensu lato, and Bartonella species in Dutch Ixodes ricinus ticks. AB - A sensitive and specific PCR hybridization assay was developed for the simultaneous detection and identification of Ehrlichia and Borrelia burgdorferi sensu lato. In separate assays the 16S rRNA gene of Ehrlichia species and the 23S 5S rRNA spacer region of B. burgdorferi sensu lato were amplified and labeled by PCR. These PCR products were used in a reverse line blot hybridization assay in which oligonucleotide probes are covalently linked to a membrane in parallel lines. Hybridization of the samples with the oligonucleotide probes on this membrane enabled the simultaneous detection and identification of Ehrlichia, B. burgdorferi, and Bartonella species in 40 different samples. The application of the assay to DNA extracts from 121 Ixodes ricinus ticks collected from roe deer demonstrated that 45% of these ticks carried Ehrlichia DNA. More than half of these positive ticks carried species with 16S rRNA gene sequences closely related to those of E. phagocytophila and the human granulocytic ehrlichiosis agent. The majority of the other positive ticks were infected with a newly identified Ehrlichia-like species. In addition, 13% of the ticks were infected with one or more B. burgdorferi genospecies. In more than 70% of the ticks 16S rRNA gene sequences for Bartonella species or other species closely related to Bartonella were found. In five of the ticks both Ehrlichia and B. burgdorferi species were detected. PMID- 10364589 TI - Comparison of the PACE 2 assay, two amplification assays, and Clearview EIA for detection of Chlamydia trachomatis in female endocervical and urine specimens. AB - Screening for sexually transmitted diseases (STDs) in a greater proportion of sexually active patients has become an accepted protocol by most health care providers. The purpose of this study was to compare the current test methods for detection of Chlamydia trachomatis used at the University of South Alabama, the PACE 2 assay (Gen-Probe) and the Clearview EIA (Wampole Laboratories), with two amplification technologies, the AMP CT (Gen-Probe) and LCx (Abbott) assays. In addition, a number of demographic parameters were ascertained by asking questions at the time of examination as well as for health care provider concerns and preferences. One urine and four endocervical swab specimens were collected in random order from 787 female patients attending one of four obstetrics-gynecology clinics. Eighty-seven percent of patients had no STD-related symptoms. Patients were considered positive for C. trachomatis if three or more assays (swab and/or urine) were positive. Abbott and Gen-Probe confirmed discrepant results by alternate amplified assays. A total of 66 true-positive specimens were detected by use of the combination of endocervical swabs and urine specimens. After discrepant analysis, sensitivities for endocervical swab specimens for the EIA and the PACE 2, LCx, and AMP CT assays were 50, 81, 97, and 100%, respectively. Sensitivities for the LCx and AMP CT assays with urine specimens were 98 and 81%, respectively. The prevalence of C. trachomatis was 8.4%, as determined by amplification technology. Overall, the amplification technologies were the most sensitive methods with either swab (AMP CT assay) or urine (LCx assay) specimens. The PACE 2 assay offered the advantage of a simpler and less expensive assay with acceptable sensitivity. The clearview CT EIA, while yielding a rapid in-office result, had unacceptably low sensitivity. The wide variation in performance with amplification assays with urine specimens as reported in both this study and the literature obviates the need to clarify optimal parameters for this specimen type. PMID- 10364590 TI - Molecular and epidemiological characterization of vaginal Saccharomyces cerevisiae isolates. AB - Although vaginitis caused by Saccharomyces cerevisiae is extremely rare, in recent years we have experienced an increasing frequency of S. cerevisiae isolation from the vaginas of fertile-age women. In order to investigate the epidemiology of these vaginal infections, a total of 40 isolates of S. cerevisiae derived from symptomatic and asymptomatic women were characterized by two DNA typing approaches, named ribosomal DNA (rDNA) hybridization and Ty917 hybridization, based on the Southern blotting technique. After transfer, the polymorphic DNA restriction fragments were hybridized with the entire repeat of S. cerevisiae rDNA for one method and with the entire sequence of the Ty917 retrotransposon for the other. After elaboration with computer-assisted analysis, the results of each method showed that Ty917 hybridization is endowed with a discriminatory power higher than that of rDNA hybridization. With the Ty917 hybridization method, all of the S. cerevisiae isolates tested appeared very heterogeneous, with the exception of those collected from individual patients with recurrent vaginitis. This allowed us to exclude a possible common source of infection while the high relatedness among S. cerevisiae sequential isolates from recurrent-vaginitis patients could suggest a pattern of relapse rather than frequent reinfection. PMID- 10364591 TI - Detection of Helicobacter pylori DNA in fecal samples from infected individuals. AB - Stool, gastric biopsy, and serum samples were collected from 22 subjects. DNA from stool was extracted, amplified, and hybridized with primers specific for the 16S rRNA gene of Helicobacter pylori. DNA from gastric biopsy specimens was analyzed similarly for comparison. Universal primers were used to confirm successful extraction of DNA from samples. Histologic, serologic, and DNA analyses were scored in a blinded fashion. Universal primer amplification verified successful DNA extraction from all stool and gastric tissue specimens. The gastric tissue DNA assay was positive for H. pylori in 11 of the 22 subjects, correlating completely with histologic and serologic results. Stool DNA was positive for H. pylori by our molecular assay in 8 of these 11 H. pylori-positive subjects. All subjects who were negative by histologic, serologic, and gastric tissue DNA analyses were also negative by stool DNA analysis. Compared to histology, serology, and gastric tissue DNA analyses, the sensitivity of our stool DNA assay was 73%, with a specificity of 100%. PMID- 10364593 TI - Usefulness of fatty acid composition for differentiation of Legionella species. AB - Numerical analysis of fatty acid methyl ester (FAME) profiles of 199 isolates and 76 reference strains, belonging to all validly described species of the genus Legionella that can be cultured in laboratory media, was used to differentiate between the species of this genus. With the exception of the strains that autofluoresced red, it was possible to differentiate all the other Legionella species. The strains of the species L. bozemanii, L. dumoffii, L. feeleii, L. gormanii, L. maceachernii, L. micdadei, and L. quinlivanii did not form single clusters, showing some degree of variability in the fatty acid compositions. The strains of the blue-white autofluorescent species had very similar fatty acid compositions and were difficult to distinguish from each other. Nine isolates had fatty acid profiles unlike those of any of the validly described species and may represent different FAME groups of known species or undescribed Legionella species. The method used in this study was useful for screening and discriminating large number of isolates of Legionella species. Moreover, the results obtained can be included in a database of fatty acid profiles, leading to a more accurate automatic identification of Legionella isolates. PMID- 10364592 TI - Validity of interpretation criteria for standardized Western blots (immunoblots) for serodiagnosis of Lyme borreliosis based on sera collected throughout Europe. AB - Western blotting (WB; immunoblotting) is a widely used tool for the serodiagnosis of Lyme borreliosis (LB), but so far, no generally accepted criteria for performance and interpretation have been established in Europe. The current study was preceeded by a detailed analysis of WB with whole-cell lysates of three species of Borrelia burgdorferi sensu lato (U. Hauser, G. Lehnert, R. Lobentanzer, and B. Wilske, J. Clin. Microbiol. 35:1433-1444, 1997). In that study, interpretation criteria for a positive WB result were developed with the data for 330 serum samples (from patients with LB in different stages [n = 189] and from a control group [n = 141]) originating mostly from southern Germany. In the present work, the interpretation criteria for strains PKo (Borrelia afzelii) and PBi (Borrelia garinii) developed in the previous study were reevaluated with 224 serum samples (from patients with LB in different stages [n = 97] and from a control group [n = 127]) originating from throughout Europe that were provided by the European Union Concerted Action on Lyme Borreliosis (EUCALB). De novo criteria were developed on the basis of the reactivities of the EUCALB sera and were evaluated with the data for the samples from southern Germany. Comparison of all results led to the following recommendations: For WB for immunoglobulin G (IgG), at least two bands among p83/100, p58, p43, p39, p30, OspC, p21, p17, and p14 for PKo and at least one band among p83/100, p39, p30, OspC, p21, and p17b for PBi; for WB for IgM, at least one band among p39, OspC, and p17 or a strong p41 band for PKo and at least one band among p39 and OspC or a strong p41 band for PBi. WB with PKo was the most sensitive, and this strain is recommended for use in WB for the serodiagnosis of LB throughout Europe. PMID- 10364594 TI - Development of amplified 16S ribosomal DNA restriction analysis for identification of Actinomyces species and comparison with pyrolysis-mass spectrometry and conventional biochemical tests. AB - Identification of Actinomyces spp. by conventional phenotypic methods is notoriously difficult and unreliable. Recently, the application of chemotaxonomic and molecular methods has clarified the taxonomy of the group and has led to the recognition of several new species. A practical and discriminatory identification method is now needed for routine identification of clinical isolates. Amplified 16S ribosomal DNA restriction analysis (ARDRA) was applied to reference strains (n = 27) and clinical isolates (n = 36) of Actinomyces spp. and other gram positive rods. Clinical strains were identified initially to the species level by conventional biochemical tests. However, given the low degree of confidence in conventional methods, the findings obtained by ARDRA were also compared with those obtained by pyrolysis-mass spectrometry. The ARDRA profiles generated by the combination of HaeIII and HpaII endonuclease digestion differentiated all reference strains to the species or subspecies level. The profiles correlated well with the findings obtained by pyrolysis-mass spectrometry and by conventional tests and enabled the identification of 31 of 36 clinical isolates to the species level. ARDRA was shown to be a simple, rapid, cost-effective, and highly discriminatory method for routine identification of Actinomyces spp. of clinical origin. PMID- 10364595 TI - Upper respiratory tract disease in the gopher tortoise is caused by Mycoplasma agassizii. AB - Upper respiratory tract disease (URTD) has been observed in a number of tortoise species, including the desert tortoise (Gopherus agassizii) and the gopher tortoise (Gopherus polyphemus). Clinical signs of URTD in gopher tortoises are similar to those in desert tortoises and include serous, mucoid, or purulent discharge from the nares, excessive tearing to purulent ocular discharge, conjunctivitis, and edema of the eyelids and ocular glands. The objectives of the present study were to determine if Mycoplasma agassizii was an etiologic agent of URTD in the gopher tortoise and to determine the clinical course of the experimental infection in a dose-response infection study. Tortoises were inoculated intranasally with 0.5 ml (0.25 ml/nostril) of either sterile SP4 broth (control group; n = 10) or 10(8) color-changing units (CCU) (total dose) of M. agassizii 723 (experimental infection group; n = 9). M. agassizii caused clinical signs compatible with those observed in tortoises with natural infections. Clinical signs of URTD were evident in seven of nine experimentally infected tortoises by 4 weeks postinfection (p.i.) and in eight of nine experimentally infected tortoises by 8 weeks p.i. In the dose-response experiments, tortoises were inoculated intranasally with a low (10(1) CCU; n = 6), medium (10(3) CCU; n = 6), or high (10(5) CCU; n = 5) dose of M. agassizii 723 or with sterile SP4 broth (n = 10). At all time points p.i. in both experiments, M. agassizii could be isolated from the nares of at least 50% of the tortoises. All of the experimentally infected tortoises seroconverted, and levels of antibody were statistically higher in infected animals than in control animals for all time points of >4 weeks p.i. (P < 0.0001). Control tortoises in both experiments did not show clinical signs, did not seroconvert, and did not have detectable M. agassizii by either culture or PCR at any point in the study. Histological lesions were compatible with those observed in tortoises with natural infections. The numbers of M. agassizii 723 did not influence the clinical expression of URTD or the antibody response, suggesting that the strain chosen for these studies was highly virulent. On the basis of the results of the transmission studies, we conclude that M. agassizii is an etiologic agent of URTD in the gopher tortoise. PMID- 10364596 TI - Detection of human papillomavirus types 6 and 11 in pubic and perianal hair from patients with genital warts. AB - Genital human papillomavirus (HPV) types 6 and 11 are of clinical importance due to their role in the development of anogenital warts. A pilot study was performed to investigate whether DNAs from HPV types 6 and 11 are present in hairs plucked from the pubic and perianal regions and eyebrows of patients with genital warts at present and patients with a recent history of genital warts. Genital HPV DNA was detected in 9 of 25 (36%) pubic hair samples and in 11 of 22 (50%) perianal hair samples by the CPI/CPIIg PCR. After sequencing of 17 of 20 samples, HPV type 6 or 11 was detected in 6 of 25 (24%) hair samples from the pubis and 8 of 22 (36%) hair samples from the perianal region. These types were not detected in plucked eyebrow hairs. In contrast, the HPV types associated with epidermodysplasia verruciformis were detected in similar proportions (62%) in both samples of pubic and eyebrow hairs. Moreover, HPV type 6 and 11 DNAs were detected in pubic hairs plucked from two patients who had been successfully treated and who did not show any lesion at the time of hair collection; this finding is an argument that HPV DNA may persist in this region. The presence of genital HPV types in plucked pubic and perianal hair suggests that there is an endogenous reservoir for HPV which may play a role in the recurrences of genital warts. PMID- 10364597 TI - Relationship between Helicobacter pylori iceA, cagA, and vacA status and clinical outcome: studies in four different countries. AB - There is continuing interest in identifying Helicobacter pylori virulence factors that might predict the risk for symptomatic clinical outcomes. It has been proposed that iceA and cagA genes are such markers and can identify patients with peptic ulcers. We compared H. pylori isolates from four countries, looking at the cagA and vacA genotypes, iceA alleles, and presentation of the infection. We used PCR to examine iceA, vacA, and cagA status of 424 H. pylori isolates obtained from patients with different clinical presentations (peptic ulcer, gastric cancer, and atrophic gastritis). The H. pylori isolates examined included 107 strains from Bogota, Colombia, 70 from Houston, Tex., 135 from Seoul, Korea, and 112 from Kyoto, Japan. The predominant genotype differed among countries: the cagA-positive iceA1 vacA s1c-m1 genotype was predominant in Japan and Korea, the cagA-positive iceA2 vacA s1b-m1 genotype was predominant in the United States, and the cagA-positive iceA2 vacA s1a-m1 genotype was predominant in Colombia. There was no association between the iceA, vacA, or cagA status and clinical outcome in patients in the countries studied. iceA status shows considerable geographic differences, and neither iceA nor combinations of iceA, vacA, and cagA were helpful in predicting the clinical presentation of an H. pylori infection. PMID- 10364598 TI - Nosocomial pseudoepidemic caused by Bacillus cereus traced to contaminated ethyl alcohol from a liquor factory. AB - From September 1990 to October 1990, 15 patients who were admitted to four different departments of the National Taiwan University Hospital, including nine patients in the emergency department, three in the hematology/oncology ward, two in the surgical intensive care unit, and one in a pediatric ward, were found to have positive blood (14 patients) or pleural effusion (1 patient) cultures for Bacillus cereus. After extensive surveillance cultures, 19 additional isolates of B. cereus were recovered from 70% ethyl alcohol that had been used as a skin disinfectant (14 isolates from different locations in the hospital) and from 95% ethyl alcohol (5 isolates from five alcohol tanks in the pharmacy department), and 10 isolates were recovered from 95% ethyl alcohol from the factory which supplied the alcohol to the hospital. In addition to these 44 isolates of B. cereus, 12 epidemiologically unrelated B. cereus isolates, one Bacillus sphaericus isolate from a blood specimen from a patient seen in May 1990, and two B. sphaericus isolates from 95% alcohol in the liquor factory were also studied for their microbiological relatedness. Among these isolates, antibiotypes were determined by using the disk diffusion method and the E test, biotypes were created with the results of the Vitek Bacillus Biochemical Card test, and random amplified polymorphic DNA (RAPD) patterns were generated by arbitrarily primed PCR. Two clones of the 15 B. cereus isolates recovered from patients were identified (clone A from 2 patients and clone B from 13 patients), and all 29 isolates of B. cereus recovered from 70 or 95% ethyl alcohol in the hospital or in the factory belonged to clone B. The antibiotype and RAPD pattern of the B. sphaericus isolate from the patient were different from those of isolates from the factory. Our data show that the pseudoepidemic was caused by a clone (clone B) of B. cereus from contaminated 70% ethyl alcohol used in the hospital, which we successfully traced to preexisting contaminated 95% ethyl alcohol from the supplier, and by another clone (clone A) without an identifiable source. PMID- 10364599 TI - Detection of equine antibodies to babesia caballi by recombinant B. caballi rhoptry-associated protein 1 in a competitive-inhibition enzyme-linked immunosorbent assay. AB - A competitive-inhibition enzyme-linked immunosorbent assay (cELISA) was developed for detection of equine antibodies specific for Babesia caballi. The assay used recombinant B. caballi rhoptry-associated protein 1 (RAP-1) and monoclonal antibody (MAb) 79/17.18.5, which is reactive with a peptide epitope of a native 60-kDa B. caballi antigen. The gene encoding the recombinant antigen was sequenced, and database analysis revealed that the gene product is a rhoptry associated protein. Cloning and expression of a truncated copy of the gene demonstrated that MAb 79/17.18.5 reacts with the C-terminal repeat region of the protein. The cELISA was used to evaluate 302 equine serum samples previously tested for antibodies to B. caballi by a standardized complement fixation test (CFT). The results of cELISA and CFT were 73% concordant. Seventy-two of the 77 serum samples with discordant results were CFT negative and cELISA positive. Further evaluation of the serum samples with discordant results by indirect immunofluorescence assay (IFA) demonstrated that at a serum dilution of 1:200, 48 of the CFT-negative and cELISA-positive serum samples contained antibodies reactive with B. caballi RAP-1. Four of five CFT-positive and cELISA-negative serum samples contained antibodies reactive with B. caballi when they were tested by IFA. These data indicate that following infection with B. caballi, horses consistently produce antibody to the RAP-1 epitope defined by MAb 79/17.18.5, and when used in the cELISA format, recombinant RAP-1 is a useful antigen for the serologic detection of anti-B. caballi antibodies. PMID- 10364600 TI - Worldwide evaluation of DNA sequencing approaches for identification of drug resistance mutations in the human immunodeficiency virus type 1 reverse transcriptase. AB - A panel (ENVA-1) of well-defined blinded samples containing wild-type and mutant human immunodeficiency virus type 1 (HIV-1) reverse transcriptase was analyzed by automated DNA sequencing in 23 laboratories worldwide. Drug resistance mutations at codons 41, 215, and 184 were present in the panel samples at different ratios to the wild type. The presence of mutant genotypes was determined qualitatively and quantitatively. All laboratories reported the presence of sequence heterogeneities at codons 41, 215, and 184 in one or more of the panel samples, though not all reported the correct codon genotypes. Two laboratories reported a mutant genotype in samples containing only the wild type, whereas two and three laboratories failed to detect the mutant genotypes at codons 41 and 215, respectively, in a completely mutant DNA population. Mutations present at relative concentrations of 25% of the total DNA population were successfully identified by 13 of 23, 10 of 23, and 16 of 23 labs for codons 41, 215, and 184Val, respectively. For more than 80% of those laboratories that qualitatively detected the presence of a mutation correctly, the estimated wild type/mutant ratio was less than 25% different from the input ratio in those samples containing 25 to 50% or 75% mutant input. This first multicenter study on the quality of DNA sequencing approaches for identifying HIV-1 drug resistance mutations revealed large interlaboratory differences in the quality of the results. The application of these procedures in their current state would in several cases lead to inaccurate or even incorrect diagnostic results. Therefore, proper quality control and standardization are urgently needed. PMID- 10364601 TI - Evaluation of mycology laboratory proficiency testing. AB - Changes over the last decade in overt proficiency testing (OPT) regulations have been ostensibly directed at improving laboratory performance on patient samples. However, the overt (unblinded) format of the tests and regulatory penalties associated with incorrect values allow and encourage laboratorians to take extra precautions with OPT analytes. As a result OPT may measure optimal laboratory performance instead of the intended target of typical performance attained during routine patient testing. This study addresses this issue by evaluating medical mycology OPT and comparing its fungal specimen identification error rates to those obtained in a covert (blinded) proficiency testing (CPT) program. Identifications from 188 laboratories participating in the New York State mycology OPT from 1982 to 1994 were compared with the identifications of the same fungi recovered from patient specimens in 1989 and 1994 as part of the routine procedures of 88 of these laboratories. The consistency in the identification of OPT specimens was sufficient to make accurate predictions of OPT error rates. However, while the error rates in OPT and CPT were similar for Candida albicans, significantly higher error rates were found in CPT for Candida tropicalis, Candida glabrata, and other common pathogenic fungi. These differences may, in part, be due to OPT's use of ideal organism representatives cultured under optimum growth conditions. This difference, as well as the organism-dependent error rate differences, reflects the limitations of OPT as a means of assessing the quality of routine laboratory performance in medical mycology. PMID- 10364602 TI - Distinct variants of Helicobacter pylori cagA are associated with vacA subtypes. AB - The diversity of the cytotoxin-associated gene (cagA) of Helicobacter pylori was analyzed in 45 isolates obtained from nine countries. We examined variation in the 5' end of the cagA open reading frame as determined by PCR and sequencing. Phylogenetic analysis revealed the existence of at least two distinct types of cagA. One variant (cagA1) was found exclusively in strains from Europe, the United States, and Australia, whereas a novel variant (cagA2) was found in strains from East Asia. The greatest diversity between cagA1 and cagA2 was found in the first 20 amino acids of the cagA open reading frame, where several consistent insertions or deletions were observed. Additional cagA sequence variants that could be classified as separate subtypes were found in two of three Peruvian and in five of seven U.S. strains tested. The calculated isoelectric point of the first 154 amino acids of the cagA1 variants (7.52 +/- 1.54) was significantly higher than that of the first 154 amino acids of the cagA2 variants (5.61 +/- 0.94; P < 0.001). Most cagA2 strains contained vacA subtype s1c (P < 0.001), and in vacA m1 strains cagA1 was more frequently observed than cagA2. These results show the epidemiological relationship between cagA and vacA at the subtype level and indicate the existence of distinct H. pylori lineages that are not uniformly distributed over the globe. PMID- 10364603 TI - Evaluation of Statens Serum Institut enteric medium for detection of enteric pathogens. AB - The efficacy of the Statens Serum Institut (SSI) enteric medium for isolation and direct identification of enteric pathogens was evaluated. Six different biochemical reactions can be read by using the SSI enteric medium, allowing direct identification of a range of enteric pathogens. All 248 gram-negative bacterial species that were tested grew on the SSI enteric medium. Only 10 of 248 bacteria (4%) showed discrepant results in the biochemical reactions, and none of these were enteric pathogens. Forty-three of 47 enteric pathogens (92%) produced identical rates of semiquantitative growth on the SSI enteric medium and 5% blood agar, whereas three Vibrio spp. and one Aeromonas spp. showed reduced growth. Gram-positive bacteria did not grow on the SSI enteric medium. Most enteric pathogens had a detection limit of 50 bacteria per ml of feces, but higher numbers of Vibrio spp. and some Shigella spp. were required for detection. The growth rates of 125 enteric pathogens and 12 Yersinia spp. on the SSI enteric medium, xylose lysine deoxycholate (XLD), Hektoen enteric (HE), Salmonella Shigella (SS), and cefsulodin-irgasan-novobiocin (CIN) agar were compared. Detection rates after application of 200 CFU were 99% for SSI enteric medium, 92% for XLD, 88% for HE, and 82% for SS agar. The 12 Yersinia spp. grew excellently on both the SSI enteric medium and CIN agar. We conclude that the performance of the SSI enteric medium compares favorably to those of other media tested. Its ability to detect Yersinia spp. may limit the number of media needed in the typical laboratory. The direct identification of enteric pathogens on the medium may also provide a more rapid diagnosis. PMID- 10364604 TI - Detection by an immunofluorescence test of Encephalitozoon intestinalis spores in routinely formalin-fixed stool samples stored at room temperature. AB - Of the several microsporidia that infect humans, Enterocytozoon bieneusi is known to cause a gastrointestinal disease whereas Encephalitozoon intestinalis causes both a disseminated and an intestinal disease. Although several different staining techniques, including the chromotrope technique and its modifications, Uvitex 2B, and the quick-hot Gram-chromotrope procedure, detect microsporidian spores in fecal smears and other clinical samples, they do not identify the species of microsporidia. A need for an easily performed test therefore exists. We reevaluated 120 stool samples that had been found positive for microsporidia previously, using the quick-hot Gram-chromotrope technique, and segregated them into two groups on the basis of spore size. We also screened the smears by immunofluorescence microscopy, using a polyclonal rabbit anti-E. intestinalis serum at a dilution of 1:400. Spores in 29 (24.1%) of the 120 samples fluoresced brightly, indicating that they were E. intestinalis spores. No intense background or cross-reactivity with bacteria, yeasts, or other structures in the stool samples was seen. Additionally, the numbers of spores that fluoresced in seven of these samples were substantially smaller than the numbers of spores that were present in the stained smears, indicating that these samples were probably derived from patients with mixed infections of Enterocytozoon bieneusi and E. intestinalis. Because a 1:400 dilution of this serum does not react with culture grown Encephalitozoon hellem, Encephalitozoon cuniculi, or Vittaforma corneae or with Enterocytozoon bieneusi spores in feces, we concluded that an immunofluorescence test using this serum is a good alternative for the specific identification of E. intestinalis infections. PMID- 10364605 TI - Nocardia thyroiditis: unusual location of infection. AB - Nocardia asteroides complex is an important opportunistic agent in immunocompromised hosts. Usually, primary pulmonary infection occurs and is followed by dissemination of the pathogen to the central nervous system and soft tissues. As described in the literature, almost every organ can be infected, but to our knowledge, Nocardia has been described as a pathogen responsible for thyroid abscess in only one report, which was published in 1993. The present report is the second case report of Nocardia thyroiditis. The patient was under immunosuppressor treatment following a combined liver-kidney transplant and presented with a preexisting nodular goiter which was probably a predisposing factor to the start and development of the thyroid infection. PMID- 10364606 TI - Prenatal diagnosis of parvovirus B19-induced hydrops fetalis by chemiluminescence in situ hybridization. AB - Parvovirus B19 can be transmitted transplacentally from the infected mother to the fetus during pregnancy, and hydrops fetalis, abortion, or stillbirth can result. In our study we explored the use of chemiluminescence in situ hybridization to detect B19 DNA on cord blood cells, amniotic fluid cells, and pleuric fluid cells from several cases of hydrops fetalis. B19 DNA was detected by using digoxigenin-labeled probes immunoenzymatically visualized with the chemiluminescent adamantil-1,2-dioxetane phenyl phosphate substrate for alkaline phosphatase. The luminescent signal emitted from the hybridized probes was detected, analyzed, and measured with a high-performance, low-light-level imaging luminograph connected to an optical microscope and to a personal computer for the quantification and localization of the chemiluminescent emission inside individual cells. PMID- 10364607 TI - Amplification of a 500-base-pair fragment from cultured isolates of Mycobacterium bovis. AB - The presence of a 500-bp fragment which amplifies a region from the genome of Mycobacterium bovis (J. G. Rodriguez, G. A. Meija, P. Del Portillo, M. E. Patarroyo, and L. A. Murillo, Microbiology 141:2131-2138, 1995) was evaluated by carrying out PCR on 121 M. bovis isolates. The M. bovis strains, previously characterized by culture and biochemical tests, were isolated from cattle in different regions of Argentina, Mexico, and Colombia. Four additional strains isolated from sea lions that belong to the M. tuberculosis complex were also included in the study. All of the isolates tested were PCR positive, rendering the expected 500-bp band and giving a correlation of 100% with previous microbiological characterization. Southern blot analysis revealed a common band of 1, 800 bp and a polymorphic high-molecular-mass hybridization pattern. The results show that this assay may be useful for diagnosis and identification of M. bovis in cattle. PMID- 10364608 TI - Molecular evidence of mother-to-infant transmission of hepatitis G virus among women without known risk factors for parenteral infections. AB - Hepatitis G virus (HGV) RNA was detected in 18 of 133 pregnant women from Tanzania without known risk factors for HGV infection and in 7 of 18 children born to HGV RNA-positive mothers. Molecular evidence of mother-to-infant transmission was obtained only for three of seven children. HGV RNA was also detected in 4 of 42 children born to non-HGV-infected women. Thus, mechanisms other than materno-filial may play an important role in HGV transmission during early childhood. PMID- 10364609 TI - Dissemination of a chloramphenicol- and tetracycline-resistant but penicillin susceptible invasive clone of serotype 5 Streptococcus pneumoniae in Colombia. AB - A national surveillance conducted in Colombia between 1994 and 1996 identified serotype 5 Streptococcus pneumoniae as the second most frequent cause of invasive disease in children younger than 5 years of age. All 43 serotype 5 isolates collected during this period were shown to be susceptible to penicillin, erythromycin, cefotaxime, and vancomycin, but most (38 of 43, or 88%) were highly resistant to chloramphenicol. In order to clarify a possible genetic relatedness among these isolates, additional microbiological and molecular characterizations were performed. Most (40 of 43, or 93%) of the isolates were found to be resistant to tetracycline. Pulsed-field gel electrophoresis (PFGE) patterns of chromosomal DNAs revealed that all the 43 isolates were closely related and that 38 of the 43 isolates were representatives of a "Colombian clone" of S. pneumoniae isolates which were recovered throughout the 3-year surveillance period from patients in 13 hospitals located in five Colombian cities. Isolates belonging to this Colombian clone were resistant to chloramphenicol and tetracycline, hybridized with the cat and tetM DNA probes in the same 340-kb SmaI fragment, and had identical PFGE patterns after both SmaI and ApaI digestions. PMID- 10364611 TI - Rapid detection of respiratory viruses by shell vial assay using simultaneous culture of HEp-2, LLC-MK2, and MDCK cells in a single vial. AB - A shell vial assay with simultaneous culture of HEp-2, LLC-MK2, and MDCK cell lines in a single tube (CoHLM SV assay) was compared with traditional tube culture (TC) for the detection of the main respiratory viruses in 358 nasal wash specimens. A total of 170 strains were isolated from 168 virus-positive samples. A total of 94. 1% of the strains (160 strains; 128 respiratory syncytial viruses and 32 other viruses) were detected by the CoHLM SV assay in 48 h, whereas 98.2% of the strains (167 strains; 132 respiratory syncytial viruses and 35 other viruses) were detected by TC in a mean time of 6 days. The CoHLM SV assay may be useful for the rapid detection of respiratory viruses. PMID- 10364610 TI - In vitro amphotericin B resistance in clinical isolates of Aspergillus terreus, with a head-to-head comparison to voriconazole. AB - Amphotericin B therapy continues to be the "gold standard" in the treatment of invasive aspergillosis in the immunocompromised host. Although Aspergillus fumigatus and Aspergillus flavus constitute the major species, several reports have described invasive pulmonary or disseminated disease due to the less common Aspergillus terreus and dismal clinical outcomes with high-dose amphotericin B. We therefore evaluated 101 clinical isolates of A. terreus for their susceptibility to amphotericin B and the investigational triazole voriconazole by using the National Committee for Clinical Laboratory Standards M27-A method modified for mould testing. Forty-eight-hour MICs indicated 98 and 0% resistance to amphotericin B and voriconazole, respectively. We conclude that A. terreus should be added to the list of etiologic agents refractory to conventional amphotericin B therapy and suggest the potential clinical utility of voriconazole in aspergillosis due to this species. PMID- 10364613 TI - Inactivation of Mycobacterium tuberculosis for DNA typing analysis. AB - DNA fingerprinting analysis of Mycobacterium tuberculosis is used for epidemiological studies and the control of laboratory cross-contamination. Because standardized procedures are not entirely safe for mycobacteriology laboratory staff, the paper proposes a new technique for the processing of specimens. The technique ensures the inactivation of M. tuberculosis before DNA extraction without the loss of DNA integrity. The control of inactivated cultures should be rigorous and should involve the use of two different culture media incubated for at least 4 months. PMID- 10364612 TI - Analysis of vacA, cagA, and IS605 genotypes and those determined by PCR amplification of DNA between repetitive sequences of Helicobacter pylori strains isolated from patients with nonulcer dyspepsia or mucosa-associated lymphoid tissue lymphoma. AB - The vacA s and m genotypes and the presence of cagA and IS605 were determined in Helicobacter pylori strains from patients with mono- and multiple infections. Surprisingly, these genetic markers were not associated with nonulcer dyspepsia or mucosa-associated lymphoid tissue lymphoma. The presence of cagA correlated with the presence of the vacA s1 allele (P < 0.05), whereas the presence of IS605 was associated with the presence of the s2 allele (P < 0.05). PMID- 10364614 TI - An untypeable Shigella flexneri strain associated with an outbreak in California. AB - Eleven Shigella flexneri (group B) isolates were recovered from epidemiologically linked patrons and food handlers from a restaurant-associated outbreak of shigellosis. Six isolates available for pulsed-field gel electrophoresis were identical. All strains agglutinated in group B and subgroup factor 6 sera but not in group 1 through group 6 sera. PMID- 10364615 TI - Clonal diversity among recently emerged strains of Vibrio parahaemolyticus O3:K6 associated with pandemic spread. AB - The genomes of the O3:K6 strains of Vibrio parahaemolyticus which abruptly emerged in Calcutta, India, in February 1996 and which demonstrated an unusual potential to spread and an enhanced propensity to cause infections were examined by different molecular techniques to determine clonality. No restriction fragment length polymorphism (RFLP) in the gene encoding the thermostable direct hemolysin was observed among the O3:K6 isolates of V. parahaemolyticus. Clonal diversity among the O3:K6 strains became evident by examining the RFLPs of the rrn operons and by the use of pulsed-field gel electrophoresis. Five ribotypes were distinguished among the O3:K6 strains examined, with ribotype R4 constituting the major type. Strains of O3:K6 isolated between June and August 1996 showed different pulsotypes compared to the pulsotypes of strains isolated before and after this period, indicating genetic reassortment among these strains, but those isolated between August 1996 and March 1998 showed identical or nearly similar pulsotypes. It is clear that there is a certain degree of genomic reassortment among the O3:K6 clones but that these strains are predominantly one clone. PMID- 10364616 TI - Molecular epidemiology of an outbreak of febrile gastroenteritis caused by Listeria monocytogenes in cold-smoked rainbow trout. AB - Febrile gastroenteritis in five healthy persons was associated with the consumption of vacuum-packed cold-smoked rainbow trout containing Listeria monocytogenes. L. monocytogenes isolates from the incriminated fish product lot and the stool samples were all of serotype 1/2a and were indistinguishable by pulsed-field gel electrophoresis employing AscI and SmaI. PMID- 10364617 TI - Level of human immunodeficiency virus DNA in peripheral blood mononuclear cells correlates with efficacy of antiretroviral therapy. AB - A novel colorimetric assay was developed and validated for accurate quantitation of human immunodeficiency virus (HIV) DNA in peripheral blood mononuclear cells (PBMCs). We tested 318 sequential samples from 56 subjects, 53 of whom were undergoing dual or triple therapy. Patients were considered responders when viremia levels were below 5, 000 HIV RNA copies/ml. The mean DNA copy numbers for untreated and responder subjects were similar (72 and 75, respectively), while it was 4.54-fold higher for nonresponders (339). This report provides strong evidence that HIV DNA levels in PBMCs correlate with therapeutic efficacy and suggests that DNA quantitation is a useful tool to monitor the decay of the HIV reservoir toward disease remission, especially when viremia is undetectable. PMID- 10364618 TI - Evaluation of an Epstein-Barr virus (EBV) immunoglobulin M enzyme-linked immunosorbent assay using a synthetic convergent peptide library, or mixotope, for diagnosis of primary EBV infection. AB - An immunoglobulin M (IgM) enzyme-linked immunosorbent assay (ELISA) was developed by using a 24-amino-acid peptide of the 18-kDa Epstein-Barr virus (EBV) viral capsid antigen (VCAp18). IgM detection was increased by 23% by using this antigen. Detection of IgM antibodies to the EBV proteins in the new ELISA was 100% specific and 95% sensitive. PMID- 10364619 TI - Delayed versus immediate bedside inoculation of culture media for diagnosis of vaginal trichomonosis. AB - A comparison of delayed versus immediate inoculation of culture medium for the diagnosis of trichomonosis was conducted. The sensitivities of the two methods were 100 and 97.4%, respectively. Delayed inoculation of culture medium for women without evidence of trichomonosis on direct microscopic examination is a valid diagnostic procedure. PMID- 10364620 TI - Molecular epidemiology of hepatitis C virus infection in an area of hyperendemicity in southern Italy: a population-based study. AB - In a cohort of subjects from Italy, anti-hepatitis C virus (HCV) and HCV RNA [HCV(+) subgroup] prevalences were 24.6 and 79.6%, respectively. HCV types 1b and 2a/c accounted for 95% of infections. Adjusted alanine aminotransferase levels were higher in males than in females and in RNA-positive subjects than in RNA negative subjects regardless of HCV type. Genotype distribution was unrelated to demographic variables. PMID- 10364621 TI - Distribution of human rotavirus G types circulating in Paris, France, during the 1997-1998 epidemic: high prevalence of type G4. AB - Group A human rotavirus G genotypes were determined by means of reverse transcription-PCR in 170 stool specimens from children with acute diarrhea admitted to a Paris children's hospital during a 1-year survey (1997 to 1998). The isolates all belonged to types G1 to G4, with type G4 predominating (60%). PMID- 10364622 TI - Evaluation of the AnaeroPack Campylo system for growth of microaerophilic bacteria. AB - Growth of microaerophilic bacteria in the AnaeroPack Campylo (Mitsubishi Gas Chemical America, Inc., New York, N.Y.) atmosphere generation system was compared to growth in the CampyPak Plus jar and CampyPak pouch (Becton-Dickinson Microbiology Systems [BDMS], Cockeysville, Md.). Growth in the AnaeroPack Campylo system was considered equivalent to or better than growth obtained in the CampyPak Plus and CampyPak pouch systems for 48 of the 50 Helicobacter pylori strains and for all 28 Campylobacter species tested. All of the 78 organisms tested were recovered in each system in equivalent colony counts. Two strains of H. pylori grown in the AnaeroPack Campylo system were observed to have colony morphology growth discrepancies when compared to growth in the two BDMS systems. Atmosphere failure with the AnaeroPack Campylo was not detected with Campylobacter jejuni ATCC 33291 used as a growth control. The AnaeroPack Campylo system is easy to use and supports the growth of campylobacters and H. pylori. PMID- 10364623 TI - Chlamydia pneumoniae in a free-ranging giant barred frog (Mixophyes iteratus) from Australia. AB - The koala biovar of Chlamydia pneumoniae was identified in lung tissue from a sick, free-ranging giant barred frog (Mixophyes iteratus) by using electron microscopy, C. pneumoniae-specific fluorescent-antibody staining, cell culture, and sequencing of the ompA, ompB and 16S rRNA genes. This is the first report of a chlamydial strain infecting both a homeotherm and a poikilotherm and only the fourth host (in addition to humans, koalas, and horses) to be naturally infected with this species of Chlamydia. The frog had severe, chronic, mononuclear pneumonia and nonregenerative anemia and pancytopenia. PMID- 10364624 TI - Typhoid fever due to Salmonella Kapemba infection in an otherwise healthy middle aged man. AB - We report the case of a patient with a Salmonella Kapemba infection, who suffered, 3 weeks after a holiday in Israel, occurrences of high fever and lower back pain for 10 days and icterus for 2 days before admission. Laboratory findings revealed a slight cholestasis and elevation of acute phase protein levels. In the blood culture a Salmonella Kapemba-type organism was cultured. The patient was afebrile for 10 days after hospitalization and then suddenly developed a temperature of 40 degrees C again. At the same time leukopenia, thrombocytopenia, and a rise of D-dimer levels were detected. The patient was admitted to the intensive care unit for a few days, because a disseminated intravascular coagulation was suspected. With magnetic resonance imaging and bone scintigraphy no osteomyelitis or abscess formation could be found. A transesophageal ultrasonography of the heart revealed no signs of endocarditis. In multiple stool cultures no salmonellas could be detected. After antibiotic treatment with ciprofloxacin the fever and lower back pain subsided, and the patient was discharged a fortnight later. This is the first reported case of typhoid fever due to the bacterium Salmonella Kapemba. PMID- 10364625 TI - Lipid emulsions in parenteral nutrition. AB - Lipid emulsions containing a physical mixture of medium and long chain triglycerides (MCT/LCT) are a well-proven concept in parenteral nutrition of critically ill patients. Having a demonstrably higher utilization rate, MCT/LCT emulsions do not impair liver function, produce less immune and no reticuloendothelial system function compromise, and do not interfere with pulmonary hemodynamics or gas exchange. A reduced content of n-6 polyunsaturated fatty acids can also be obtained by using newer preparations based on structured triglycerides or olive oil. Further studies are necessary in order to investigate these new lipid emulsions versus the physical mixture of MCT/LCT. A promising substrate in the development of lipid emulsions can be seen in fish oils. With regard to current literature, fish oils have a beneficial influence on the pathophysiological response to endotoxins and exert important modulations on eicosanoid and cytokine biology. Furthermore their intravenous use may improve organ perfusion in different critical situations. PMID- 10364626 TI - Macronutrient intake of 3- to 36-month-old German infants and children: results of the DONALD Study. Dortmund Nutritional and Anthropometric Longitudinally Designed Study. AB - The intake of macronutrients (protein, fat, fatty acids, carbohydrates, added sugars, fiber) was assessed in 354 healthy German infants and children aged 3-36 months from 3-day weighed diet records. The intake of protein ranged between 7 and 14% of energy intake. Fat intake decreased from 3 months (breast-fed boys and girls, 48%; formula-fed boys/girls, 41/44%) to 12 months (boys/girls, 33/36%) due to the increasing consumption of commercial weaning foods, and then increased again up to 36 months (boys/girls, 40/43%). Intake of added sugars decreased during the first 12 months and then increased again, but only slightly exceeded the limit of 10%. Intake of dietary fiber was highest at the age of 1 year (boys/girls, 2.7/2.3 g/MJ). The macronutrient intake was in accordance with other German and European surveys, but deviated considerably from the respective recommendations. PMID- 10364627 TI - Hormonal, lifestyle, and dietary factors in relation to leptin among elderly men. AB - BACKGROUND: Leptin, the adipocyte-secreted protein product of the ob gene, has been strongly linked to obesity and is believed to play a role in the regulation of the reproductive system. This study examines the potential influence of lifestyle and dietary factors, as well as of other hormones, on serum levels of leptin. METHODS: The authors studied a population of 48 healthy elderly Greek men. Sera from these men were analyzed for leptin, several steroid hormones, sex hormone-binding globulin, and insulin-like growth factor 1. The authors also utilized data from food frequency questionnaires and information on demographic, anthropometric, and lifestyle (cigarette smoking, alcohol and coffee drinking) factors. RESULTS: Using linear regression modeling, serum leptin levels were inversely associated with testosterone and positively associated with estradiol and dehydroepiandrosterone sulfate, after adjustment for the other hormones and body mass index (BMI). Leptin levels in men with a BMI >30 kg/m2 were 170% higher than in men with a BMI <27 kg/m2 (95% CI 63- 346%). Height was also positively associated with leptin, independent of BMI. No notable relationships were observed between leptin, on the one hand, and smoking, alcohol drinking, coffee drinking, or total energy intake, on the other. When total energy intake was separated into its three major components (carbohydrate, fat, and protein), it appeared that fat intake may have an isocalorically differential effect on serum leptin levels; one marginal quintile increase in fat intake corresponded to an 11% increase in leptin (95% CI 0-24%). CONCLUSION: Serum levels of leptin may be influenced by other endocrine factors, especially testosterone and estradiol, and may be positively associated with excess fat intake independently of obesity. PMID- 10364628 TI - Serum and urine selenium concentrations in patients with cardiovascular diseases and relationship to other nutritional indexes. AB - Serum and urine selenium levels were determined in patients with cardiovascular diseases by hydride generation atomic absorption spectrometry. Mean serum Se concentrations measured in patients with acute myocardial infarction (AMI; n = 32) or with ischemic cardiomyopathy (n = 50) were significantly lower than those determined in control groups. In AMI patients, serum triglyceride levels showed a positive significant correlation with the serum Se concentration (r = 0.59, p < 0.05). This result reinforces the important role of Se as an antioxidant agent in this disease. Mean urine Se concentrations of AMI patients (n = 33) were also significantly lower to those determined in the control group (p < 0. 05). This reaction of the organism contributes to regulate the Se homeostasis to keep the body Se status as high as possible. PMID- 10364629 TI - Influence of calcium intake on gestational hypertension. AB - Calcium intake during the third trimester of pregnancy was determined in 82 pregnant women by recording the consumption of foods over a 5-day period and by calculation of the quantity of this element provided by dietary supplements. For each subject, blood pressures were measured once per week using an aneroid sphygmomanometer, to detect and analyze differences in calcium intake between those with normal blood pressure and those suffering from gestational hypertension (7.3%). Calcium intake was significantly lower amongst subjects with high blood pressure (757.7 +/- 154.5 compared to 986.4 +/- 502.3 mg/day in normotensive subjects). The relationship between calcium intake and blood pressure was independent of other variables such as body mass index, number of previous pregnancies, weight gain, subject age or hematocrit levels. Though further investigation is needed, the results obtained seem to support the idea that pregnant women should try to maintain an optimal calcium intake. PMID- 10364630 TI - Effect of cholesterol feeding on DNA damage in male and female Syrian hamsters. AB - Cholesterol oxides are cytotoxic and have been implicated in many disease processes; however, it has been proposed that cholesterol oxides result from cholesterol acting as a sacrificial antioxidant. In this study, the effect of dietary cholesterol on DNA damage, assessed by the alkaline comet assay, was examined in male and female Syrian hamsters. Animals were fed ad libitum a modified AIN-76 diet (control) or a diet with 0.5% cholesterol for 10 weeks. Following the 10-week feeding period, there was no significant difference in body weight between cholesterol-fed and control animals. Cholesterol feeding resulted in significant liver hypertrophy, and increased plasma total and HDL cholesterol in both male and female animals compared with controls. There was no difference in liver cell DNA damage levels as measured by the comet assay. Heart cells from cholesterol-fed hamsters, however, showed a significant decrease in tail DNA (p = 0.050) indicating decreased damage compared with controls and a possible protective effect of cholesterol against DNA damage. PMID- 10364631 TI - Effects of caffeine on the bones of aged, ovariectomized rats. AB - Caffeine is a substance which many people consume in their daily life. Caffeine's effects on bone are still controversial. Using ovariectomized rats, the present study was conducted to determine to what extent caffeine intake affects the mechanical properties, bone minerals and histology. Aged rats were divided into 2 groups after ovariectomy. Group 1 was fed a 20% protein diet as a control, and group 2 was fed a 20% protein diet supplemented with caffeine (2 mg/100 g body weight). The respective diets were fed to the rats of each group for 90 days. Rats were then killed by heart puncture, blood was collected, and femurs were removed. In 1 group of femurs paraffin cross-sections were made at the midshaft of each bone. Total width, cortical width, total cross-sectional bone area of the midshaft, and the number of osteocytes in randomly selected areas were measured. Another group of bones was subjected to three-point bending testing until failure. Bones were then pulverized and Ca, P, Mg, Zn, Sr, Si, hydroxyproline and hexosamine contents and crystallite size were measured. Various mechanical properties, except modulus of elasticity, in the caffeine group were consistently 7-23% lower than the noncaffeine controls. Yield strain in the caffeine group was significantly less than in the noncaffeine controls. Zinc, Sr, and crystallite size of bone showed a significant decrease in the caffeine group, whereas Si contents significantly increased. Our current results indicate that routine intake of caffeine in the elderly should be regarded with some caution. PMID- 10364632 TI - Effects of guar gum and cellulose on cecal enzyme activity and cecal short-chain fatty acids in young and aged mice. AB - The effects of cellulose or guar gum on cecal enzyme activity and cecal short chain fatty acids (SCFAs) in young and aged mice were studied. Male Crj:CD-1 (ICR) mice were fed an MF diet for 4 (young mice) or 23 months (aged mice). The MF diet was then replaced with a semisynthetic diet supplemented with 5% guar gum or 5% cellulose. The mice were fed the guar gum or cellulose diet for 3 weeks. There was no significant difference in cecal content between the two diet groups. There were no significant differences in total short-chain fatty acid production between the young mice fed the cellulose and those fed the guar gum diet, and between the aged mice fed the cellulose and guar gum diet. There were significant differences in cecal enzyme activity between the young mice fed the cellulose and those fed the guar gum diet. Beta-glucuronidase activity was significantly higher in the young mice fed the guar gum diet than in those fed the cellulose diet. There were also significant differences in cecal enzyme activity between the aged mice fed the cellulose diet and those fed the guar gum diet. Beta-glucuronidase activity was significantly higher in the aged mice fed the guar gum diet than in these fed the cellulose diet. Beta-glucosidase activity was significantly lower in the aged mice fed the guar gum diet than in those fed the cellulose diet. The effect of cellulose on the microflora between the young and aged mice might be different from the effect of guar gum. The degree of adaptation to the diet of microflora in young and aged mice fed the cellulose diet might differ from that in those fed the guar gum diet. The higher enzyme activities of microflora in aged animals compared to young animals, might have some relation with the incidence of colon cancer in aged animals. PMID- 10364633 TI - Erythropoietin blood level is increased in sudden infant death. AB - Erythropoietin (EPO) is the main red cell growth factor and its release into the blood stream is stimulated by anemia and also by various kinds of hypoxia. We studied the blood EPO concentration in a population of 96 infants who died suddenly and compared their mean EPO levels to control infants. The normal values were low at birth and progressively increased during the first 2 years. In the sudden infant death (SID) group the EPO level was significantly higher (p = 0.001) for the entire population and particularly in the youngest group (0-2 months): 14.7 +/- 2.4 IU/l (mean +/- SEM) in SID group vs. 3.6 +/- 0.4 IU/l in control group (p < 0.001). Although we could not analyze the blood hemoglobin concentration after death, the anemia hypothesis was refuted by an assay of the percentage of fetal hemoglobin which was normal for age in the control and SID groups. Moreover, there was no significant difference in EPO levels between explained and unexplained deaths. We also observed an increase in the stress hormones, cortisol and beta-endorphin, in the entire SID group. These SID results suggest a profound and long-lasting hypoxia at least during terminal agony. PMID- 10364634 TI - In vivo measurements of glucose absorption in preterm infants. AB - Few data are available regarding the kinetics of glucose absorption in the preterm infant. In order to ascertain the kinetics of glucose absorption in the premature infant and how rate of infusion and concentration affect absorption, jejunal glucose absorption kinetics were measured in 16 preterm infants by infusing 1, 10, and 100 mM glucose solutions in random order. Seventeen infants were perfused with glucose (100 micromol/min) by infusing 67 mM glucose at 1.5 ml/min and 133 mM glucose at 0.75 ml/min to determine the effect of changes in rate of infusion vs. concentration on glucose absorption. Km and Vmax were related to postnatal age. Km was correlated inversely with the percentage of feedings received as human milk. Antenatal administration of glucocorticoids appeared to increase Vmax. The higher infusion rate resulted in greater glucose absorption than the higher concentration glucose solution. CONCLUSIONS: The characteristics of glucose absorption in the preterm infant change with age and are affected by diet, glucocorticoids, and the method of infusion. These data have implications for the feeding of preterm infants. PMID- 10364636 TI - Gestational magnesium deficiency is deleterious to fetal outcome. AB - A number of recent epidemiological findings have implicated magnesium as being essential to fetal well-being. Few studies, however, have examined the relationship between maternal requirements for dietary magnesium and subsequent mortality and morbidity in offspring. The present study uses a rodent model of dietary-induced hypomagnesemia to investigate the effects of magnesium deficiency prior to and during gestation on neonatal morbidity and mortality. Magnesium deficiency during gestation significantly increased neonatal mortality and morbidity. Such increases were associated with a reduced free magnesium concentration in both maternal and offspring blood and an increased incidence of periventricular hemorrhage and edema in newborn pups as observed by magnetic resonance imaging and histology. Animals fed a magnesium-deficient diet before mating but given magnesium supplementation during gestation did not demonstrate a significant change in neonatal mortality and morbidity when compared to control animals. The significant improvement in fetal outcome with dietary magnesium supports the concept of magnesium supplementation during pregnancy. PMID- 10364635 TI - Labeled trimethyllysine load depletes unlabeled carnitine in premature infants without evidence of incorporation. AB - 6-N-Trimethyl-[d9]-L-lysine (dTML), the labeled form of a mammalian carnitine precursor, was administered to two groups of premature infants. Although the urinary output of dTML significantly increased in the low-dose-treated group (100 micromol/day), this amount did not affect the urinary output or plasma levels of carnitine and carnitine esters. In the second group of infants, after administration of 500 micromol dTML the plasma-free carnitine concentration increased (from 9.95 +/- 0.63 to 12.9 +/- 0.87 nmol/ml, p > 0.05) with a significant increase in the urinary excretion of free carnitine on the day of dTML administration and on the posttreatment day (from 4.79 +/- 1.36 to 9.85 +/- 1.18 and to 17.5 +/- 2.31 micromol/day, respectively). Analysis of urine using fast atom bombardment mass spectrometry (FAB-MS) revealed only the presence of the dTML in the urine of the newborns; no change was detected in the relative abundance of any other carnitine precursor. Surprisingly, in the second group, which received the higher dose of dTML supplement, only the signal intensity of the unlabeled carnitine increased after dTML administration; no new peak appeared in the urine that would correspond to the de novo synthesized carnitine containing the stable isotope-labeled trimethyl group of dTML. Thus, the FAB-MS analysis clearly demonstrated that contrary to the likely prediction, the 270% extra free carnitine output was a consequence of a dose-dependent dTML-induced depletion of the free carnitine reserves from the newborns. The absence of the incorporation of the label from dTML into carnitine strongly suggests that circulating TML is not the precursor of carnitine in premature infants. PMID- 10364637 TI - Characteristic transfer of colostral components into cerebrospinal fluid via serum in neonatal pigs. AB - In order to evaluate the possibility of modification of brain function by colostral suckling, the characteristic transfer of colostral components into serum and cerebrospinal fluid (CSF) has been studied by SDS electrophoresis, immunoblot and ELISA methods in nonsuckling pigs. Total protein concentrations in the serum increased immediately after oral administration of bovine colostrum, reaching a peak value (7.0 +/- 0.7 g/dl) at 24 h after administration, corresponding to a 3-fold increase compared to preinfusion levels. IgG and other macromolecular components (MW 19, 000-58,000) were recognized in serum by electrophoretic and ELISA analysis. Total protein concentrations in the CSF collected from the cisterna magna also increased steeply after colostral administration, reaching a maximal value (54.1 +/- 5.0 mg/dl) at 4 h, corresponding to a 4-fold increase compared to preinfusion levels. Two colostral components (MW 19,000 and 31,000) in serum were confirmed to be present in the CSF by electrophoresis. The component of MW 19,000 was identified by immunoblot as beta-lactoglobulin. IgG in serum transferred from colostrum could not be detected in the CSF by ELISA. Lactoferrin administered into the intestine was also detected in the CSF via serum. These results indicate that some components of colostrum can be transported into the CSF via the serum, suggesting the possibility of modification of immature brain functions by colostral suckling in neonatal pigs. PMID- 10364638 TI - Adrenocortical imprint in the fetus of a diabetic gestation. AB - Offspring of diabetics are at increased risk for diabetes as adults. As corticosteroids are intimately involved in glucose homeostasis, we investigated aspects of corticosteroid activity in the late gestation fetuses of control, moderately diabetic and insulin-controlled streptozotocin-induced diabetic rats. We found that moderate maternal diabetes had no effect upon litter size or fetal body weight. Uncontrolled maternal diabetes was accompanied by fetal hyperglycemia, hyperinsulinemia and elevated aldosterone. Maternal insulin treatment normalized fetal glucose and aldosterone; fetal insulin and corticosterone levels increased. Maternal diabetes had no effect upon fetal adrenal expression of P450scc mRNA; the abundance of P450c11beta mRNA increased, and returned to that of the control gestation upon insulin treatment. P450c11AS mRNA was barely detectable, and decreased in the fetuses of insulin-treated diabetics. P450c11B3 mRNA was undetectable in all fetal groups. Our results implicate aspects of maternal diabetes in the expression of a fetal adrenocortical imprint, manifested as a greater abundance of P450c11beta mRNA. Although not accompanied by elevated corticosterone in the fetus, this imprint could ultimately allow for greater potential corticosterone production in response to typical stimuli, and thus contribute to the tendency towards glucose dysregulation in these offspring of diabetic gestations. PMID- 10364639 TI - Functional role of sialic acid in IgG binding to microvillus membranes in neonatal rat intestine. AB - A close parallelism exists between sialylation and endocytotic activity of the small intestine during postnatal development in rats. Thus, the binding of 125I labelled IgG to microvillus membranes and its relationship to membrane sialic acid has been studied in suckling rat intestine, during (a) postnatal development; (b) cortisone-induced precocious maturation, and (c) after desialylation of brush borders by neuraminidase treatment. Neuraminidase-treated membranes exhibited low (42%, p < 0.001) IgG binding. The observed decrease in IgG binding to desialylated membranes was associated with a decrease in the value of affinity constant, (-Ka = 0.4 x 10(6) M-1 in control and 0.23 x 10(6) M-1 in desialylated membranes). The number of IgG-binding sites (2.3 nmol/mg protein) was unchanged under these conditions. A similar decrease (50%) in IgG binding to brush borders was also observed in cortisone-injected pups. This was associated with reduced sialic acid (37%) content of the membranes compared to the controls. The value of -Ka was reduced from 0.4 x 10(6) M-1 in the control to 0.3 x 10(6) M 1 in the hormone-injected pups. The number of binding sites (n) was decreased from 2.2 to 1.4 nmol/mg protein under these conditions. Low concentrations of calcium (0.1-1.6 mM) in the incubation medium enhanced IgG binding (p < 0.001) to brush borders in pups but there was no change in binding of IgG to the membranes at 2 mM Ca2+ concentration compared to controls. Addition of Zn2+ or Mg2+ did not affect IgG binding under these conditions. These findings suggest a functional role of Ca2+ and sialic acid residues of the membrane glycans in IgG-receptor interactions in suckling rat intestine. PMID- 10364640 TI - The role of the polymorphic genes apolipoprotein E and methylene- tetrahydrofolate reductase in the development of dementia of the Alzheimer type. AB - The gene for apolipoprotein E (APOE) is polymorphic, and its variant APOE4 is a major risk factor for the development of Alzheimer-type dementia (AD). Another risk factor for AD appears to be negative cobalamin balance, which is very common in elderly people. Cobalamin and folate are interdependent and essential components of the one-carbon metabolism. Another important component is methylenetetrahydrofolate reductase (MTHFR), the gene for which is also polymorphic. Thermolabile MTHFR (tMTHFR), a gene variant that reduces the activity of its enzyme, is common in the general population. In the present study, 75% of 140 AD patients had at least one APOE4 allele. The numbers of APOE4 and tMTHFR alleles correlated significantly with the serum folate levels, however, in opposite directions. The significance of this was augmented by an inverse correlation between APOE4 and tMTHFR. Thus, not only MTHFR but also APOE appears to be related to the one-carbon metabolism, suggesting that APOE4 and insufficient one-carbon metabolism may be synergistic risk factors for AD. PMID- 10364641 TI - Astrogliosis and the ApoE genotype. an immunohistochemical study of postmortem human brain tissue. AB - Significantly increased glial fibrillary acidic protein (GFAP) expression was seen in postmortem brain tissue from demented patients with Alzheimer's disease (AD) (73 cases, 61 females/12 males, mean age 84 +/- 9 years) compared to controls (22 cases, 10 females/12 males, mean age 78 +/- 9 years). In demented patients, the GFAP expression, counts of AD lesions, senile/neuritic plaques (SP/NP) and neurofibrillary tangles (NFT) were higher in patients carrying the apolipoprotein E (ApoE) epsilon4 allele compared to those without the ApoE epsilon4 allele. The astrogliosis correlated significantly with the counts of NFT in demented patients both with and without the ApoE epsilon4 allele. Furthermore, the astrogliosis correlated significantly with the counts of SP/NP, but this correlation, however, was influenced by the ApoE epsilon4 allele, being significant only in those cases without the ApoE epsilon4 allele. Our results demonstrate that not only the extent of AD lesions and GFAP expression, but also the relationship between AD lesions and the GFAP expression is influenced by the ApoE epsilon4 allele. PMID- 10364642 TI - Early-onset and late-onset depression are independent of the genetic polymorphism of apolipoprotein E. AB - The recently shown association between apolipoprotein E (APOE) genotype and depressive illness has been challenged by subsequent studies. However, controversial results may derive from the different diagnostic criteria used for depression and from the small numbers of depressed patients included in the studies. We examined the association between depression and the genetic polymorphism of APOE in a large sample of depressed patients, Alzheimer's disease (AD) patients, and healthy controls following clear definitions for late-life depression. The cumulative incidence of depression depending on the age at onset of the first episode was examined by survival analysis. Our data do not disconfirm the hypothesis of depression sharing some common pathophysiologic features with AD, however, it seems very unlikely that the APOE genotype will elucidate the assumed common mechanisms. PMID- 10364643 TI - Inheritance of the ApoE epsilon4 allele increases the rate of brain atrophy in dementia patients. AB - We investigated the influence of the apolipoprotein (ApoE) epsilon4 allele on the rate of brain atrophy in patients with clinical dementia and in subjects at risk for dementia. Eighty-one subjects, consecutively referred to a memory clinic due to symptoms of dementia, went through a comprehensive examination, including cerebral magnetic resonance imaging. After an initial investigation these subjects were divided into one of six diagnostic groups; Alzheimer's disease (AD, n = 23), objective cognitive impairment (OCI, n = 27), subjective cognitive impairment (SCI, n = 17), vascular dementia (VaD), frontotemporal dementia (FTD) and unspecified dementia (USD). The last three groups were joined into one diagnostic group designated 'other dementia' (OD, altogether n = 14). In order to study the progression of cognitive impairment as well as the rate of atrophy in different brain regions all subjects were reinvestigated after an average period of 16 months. Interest was focused on investigating if those subjects with one or two epsilon4 alleles differed in either dementia progression or rate of brain atrophy compared to those without the epsilon4 allele. We found that the ApoE epsilon4 carriers had a statistically significantly larger increase in ventricular volume as compared with the ApoE epsilon4 noncarriers. In all diagnostic groups the ApoE epsilon4 carriers showed a greater rate of ventricular volume increase, as compared to the noncarriers. However, this difference was statistically significant only for the OD subjects. No statistical significant changes over time were seen for whole brain volume or volume of the temporal lobes and the medial temporal lobes. The diagnostic groups differed in dementia progression with the AD subjects having the most pronounced reduction in MMSE scores as compared to subjects at risk for AD (OCI and SCI subjects). The presence of ApoE epsilon4 allele did not influence the change in MMSE in any of the diagnostic groups. PMID- 10364644 TI - Validity of different linguistic versions of the Alzheimer's Disease Assessment Scale in an international multicentre Alzheimer's disease trial. AB - Owing to the involvement of Italian Centres in a multicentre, German-Italian therapeutical trial with Alzheimer's dementia patients, to be assessed with the Alzheimer's Disease Assessment Scale (ADAS), it was decided that the Italian centres would use an Italian version of the scale, derived from that used by the German centres. However, the lists of words for exploring verbal memory are not merely translated from the German version, but are composed of selective Italian words chosen according to linguistic criteria. This Italian version was validated following the same procedure adopted for validating the German version. We submitted this Italian version to an interrater reliability, test-retest reliability, concurrent validity, internal consistency and sensitivity evaluation, using demented patients. Based on the results of these tests this Italian version of the ADAS proved valid and reliable. Moreover, the results were strikingly comparable to those from the validation of the German version. Our work supports the validity, reliability and transnational comparability of national versions of the ADAS constructed following definite linguistic criteria. PMID- 10364645 TI - Occurrence of delirium in different regional brain syndromes. AB - Old age and organic brain disorder are major risk factors for delirium in the elderly. The localization of the brain damage in dementia may be of importance for this risk. The aim of the present study was to investigate the rate of delirium in patients with different regional brain syndromes. Included in the study were 194 demented patients. The occurrence and intensity of four regional brain syndromes were estimated: parietal lobe syndrome, frontal lobe syndrome, subcortical syndrome and global brain syndrome. The analyses showed that the rate of delirium was significantly higher when a global brain syndrome was predominant than when any one of the other syndromes predominated. PMID- 10364646 TI - Magnetic resonance imaging-based hippocampal volumetry in patients with dementia of the Alzheimer type. AB - A volumetric magnetic resonance imaging assessment of the hippocampal region was done in 50 patients with Alzheimer-type dementia and 25 members of a control group. There were no significant volumetric differences between the right and left hippocampi. The volumetric hippocampal measurements were 39.4% smaller in dementia of the Alzheimer type than in controls. In the differentiation between the analyzed groups of patients, the method had a sensitivity of 95% and a specificity of 92%. A correlation of the hippocampal volumes with the severity of the dementia and the patient's age was found. The results confirmed the importance of hippocampal atrophy in the etiology of dementia. In vivo the quantitative assessment of the hippocampal formation is a valuable tool in the diagnosis of Alzheimer's disease. PMID- 10364647 TI - A double-blind, placebo-controlled study of tacrine in Chinese patients with Alzheimer's disease. AB - The purpose of the study was to evaluate the efficacy and safety of tacrine over 30 weeks in Chinese patients with probable Alzheimer's disease (AD). A total of 100 patients with mild to moderate AD were recruited and randomly assigned to active or placebo treatment. The active group received 30 mg/day of tacrine for the first 6 weeks, 60 mg/day for the next 6 weeks, 90 mg/day for 6 more weeks and then 120 mg/day for the remaining 12 weeks. Safety evaluations included biweekly determinations of alanine aminotransferase (ALT). The primary outcome measures were Cognitive Abilities Screening Instrument (CASI), Clinical Global Impression of Change (CGIC) by investigator and the Informant Questionnaire on Cognitive Decline in the Elderly (IQCODE). Secondary outcome measures were Mini-mental State Examination (MMSE), Alzheimer's Deficit Scale (ADS) and CGIC by caregivers. Sixty-eight patients were included in an intent-to-treat analysis (48 active and 20 placebo); 56 patients had evaluable data at week 30 (36 active and 20 placebo). The results of the complete case analysis revealed a significant improvement in the CASI and MMSE scores of the active group in the 18th week (90 mg/day) and the 30th week (120 mg/day) (p < 0.01). In the intent-to-treat analysis, significant improvement of the active group was noted on CASI at week 30 (p = 0.05), but there was no significant difference in the measures of IQCODE, CGIC and ADS. The primary reasons for withdrawal of tacrine-treated patients (39 patients, 52%) were asymptomatic ALT elevation, anorexia and nausea/vomiting. These patients all recovered from the adverse events on discontinuation of treatment. Tacrine produced a statistically significant improvement in the CASI and MMSE in Chinese patients with mild to moderate AD using a lower dose than in western people. PMID- 10364648 TI - The cost of dementia in Denmark: the Odense Study. AB - In a population-based study of dementia, the cost of care for 245 demented elderly and 490 controls matched by age and gender was estimated. Dementia of Alzheimer's type was diagnosed according to the NINCDS-ADRDA criteria, and vascular dementia and other types of dementia were diagnosed according to the DSM IIIR criteria. Severity of dementia was determined by the Clinical Dementia Rating scale. The annual cost of medical care, domestic care, home help, nursing home and special equipment for nondemented patients was DKK 22,000 per person while the cost for very mildly, mildly, moderately and severely demented patients was DKK 49,000, DKK 93,000, DKK 138,000 and DKK 206,000, respectively. Except for very mild dementia the cost did not differ between elderly who suffer from Alzheimer's disease and those with other types of dementia. The net cost of dementia is the difference in cost between those with dementia and the matched controls and amounts on average to DKK 77,000 per person per year. However, priority setting cannot be based on the cost of dementia per se, but only on the cost of a specific dementia intervention compared to its health benefit. PMID- 10364649 TI - A meta-analysis on the efficacy and tolerability of alpha1-adrenoceptor antagonists in patients with lower urinary tract symptoms suggestive of benign prostatic obstruction. AB - OBJECTIVE: To assess whether the alpha1-adrenoceptor antagonists currently available for the treatment of lower urinary tract symptoms (LUTS) suggestive of benign prostatic obstruction (BPO) (alfuzosin, terazosin, doxazosin and tamsulosin) can be distinguished with regard to clinical efficacy and/or tolerability. METHODS: Up-to-date analysis of clinical placebo-controlled or direct comparative studies with alpha1-adrenoceptor antagonists in patients with LUTS suggestive of BPO derived from a MEDLINE search in October 1998. All retrieved studies were analyzed with regard to efficacy and tolerability. Efficacy was evaluated by the percentage improvement in total symptom score and Qmax (mean end of study value relative to mean baseline value). Tolerability was evaluated by means of study withdrawal rate because of adverse events and the incidence of vasodilatatory adverse events (e.g. dizziness and orthostatic hypotension). RESULTS: Indirect comparison of data derived from the placebo controlled studies involving 6,333 patients and the data derived from the direct comparative studies involving 507 patients demonstrate that all alpha1 adrenoceptor antagonists (alfuzosin, terazosin, doxazosin and tamsulosin) produce comparable improvements in LUTS and urinary flow. Total symptom score is in general improved by 30-40% and Qmax by 16-25%. The difference between currently available alpha1-adrenoceptor antagonists is related to their side effect profile. Alfuzosin (especially the sustained release formulation) and tamsulosin (modified release formulation 0.4 mg) seem to be better tolerated than terazosin and doxazosin. The percentage of patients that withdrew due to bothersome side effects with alfuzosin and tamsulosin 0.4 mg was comparable to that with placebo (about 4-10%) whereas in the terazosin and doxazosin studies an additional 4-10% of patients dropped out because they did not tolerate the therapy. Tamsulosin has less effect on blood pressure than alfuzosin (especially in elderly patients) and causes less symptomatic orthostatic hypotension during orthostatic stress testing than terazosin. CONCLUSIONS: All alpha1-adrenoceptor antagonists seem to have similar efficacy in improving symptoms and flow. The difference between alpha1 adrenoceptor antagonists is related to their side effect profile. Alfuzosin and tamsulosin appear to be better tolerated than doxazosin, terazosin and prazosin. PMID- 10364650 TI - Laparoscopic radical prostatectomy: technical and early oncological assessment of 40 operations. AB - OBJECTIVE: To evaluate the technical feasibility, oncological efficacy and intraoperative and postoperative morbidity of laparoscopic radical prostatectomy. METHOD: We describe an original technique of laparoscopic radical prostatectomy performed in 40 patients between 26th January and 12th October, 1998. RESULTS: Radical prostatectomy was performed entirely by laparoscopy in 35 patients (87.5%) and only one conversion was performed in the last 26 patients (4%). Pelvic lymphadenectomy was performed in the light of preoperative staging data in 14 patients (35%). The median total operating time was 270 min. The only major complication was a rectal injury (patient 8), sutured laparoscopically with an uneventful postoperative course. Postoperative vesical catheterization lasted an average of 7.65 days. Seven patients were transfused (17.5%) with an average of 2.8 units of packed cells (range: 2-3). The reduction of postoperative pain is an element allowing for a rapid discharge of the patients by the 3rd postoperative day. The oncological results were as follows: 36 patients had a pT2 tumor (90%); prostate tumor was staged as N0 in 14 cases and NX in 26 cases. Surgical margins were negative in 33 patients (82.5%). Two patients had a doubtful resection margin (1 at the apex and 1 at the bladder neck) and 5 patients had positive margins. The last PSA level was undetectable (<0.1 ng/ml) in 26 (89.7%) of the 29 patients in whom PSA level was available more than 1 month after the operation. Functional results are not yet available and will be published later. CONCLUSIONS: Radical prostatectomy is an operation which can be routinely performed by laparoscopy by a team experienced with this technique. Operative and postoperative morbidity was low. Short-term oncological data appear identical to the results of conventional retropubic surgery. The improvement of operative visibility was considerable allowing a much more precise dissection. The laparoscopic approach appears to represent a technical improvement of the radical prostatectomy if the functional results of this operation improve parallel to the quality of dissection. A long-term follow-up is needed to define definitively the place of this new approach to radical prostatectomy. PMID- 10364652 TI - Fluorine-18-fluorodeoxyglucose positron emission tomography is useless for the detection of local recurrence after radical prostatectomy. AB - OBJECTIVE: After radical retropubic prostatectomy a rise of the prostate-specific antigen (PSA) indicates a local recurrent or metastatic disease. If the bone scan shows no apparent bone metastasis, morphological imaging methods like x-ray computed tomography, magnetic resonance imaging or transrectal ultrasound often cannot distinguish between postoperative scar and local recurrence. Therefore we investigated the feasibility of fluorine-18-fluorodeoxyglucose positron emission tomography (F-18 FDG PET) for metabolic characterization of prostatic cancer, especially for differentiation of scar or recurrent prostate cancer after radical prostatectomy. METHODS: Dynamic PET with 370 MBq F-18 deoxyglucose (F-18 FDG) up to 60 min p.i. was performed in 2 patients with biopsy-proven benign prostatic hyperplasia, in 11 patients with a histologically proven prostate cancer prior to radical retropubic prostatectomy (RRP) and 7 patients with suspected local recurrence (with negative bone scan) after RRP prior to biopsy of anastomosis (3 local recurrence, 4 postoperative scar). RESULTS: Prostate cancer showed a very low F-18 FDG uptake. The placement of regions of interest was only possible by the use of other imaging methods. There was not difference between the F-18 FDG uptake of benign prostate hyperplasia, prostate carcinoma, postoperative scar or local recurrence after radical prostatectomy. CONCLUSION: F-18 FDG seems not to be useful to distinguish between postoperative scar and local recurrence after radical prostatectomy. PMID- 10364651 TI - Early prostate-specific antigen relapse after radical retropubic prostatectomy: prediction on the basis of preoperative and postoperative tumor characteristics. AB - OBJECTIVES: This study was undertaken to distinguish between patients who will and will not benefit from a retropubic radical prostatectomy (RRP) for clinically localized prostatic carcinoma (PCa) on the basis of preoperative and postoperative tumor characteristics. METHODS: Data of 318 consecutive patients who underwent RRP for clinically localized PCa were reviewed. Preoperative characteristics used included clinical stage, findings on transrectal ultrasonography, prostate-specific antigen (PSA) values, Gleason grade, number of positive biopsies, number of biopsies containing any Gleason grade 4 and/or 5 cancer, and number of biopsies with predominant (>50% of cancerous tissue) Gleason grade 4 and/or 5 cancer. Postoperative characteristics included pathologic stage, Gleason grade, margin status, cancer volume, and volume of Gleason grade 4 and/or 5 cancer. The impact on biochemical relapse after RRP were calculated by Cox regression and CART (classification and regression tree) analysis to establish low, intermediate, and high risk of recurrence. RESULTS: Of patients who underwent RRP, 66% showed no evidence of relapse after a follow-up of 42 months. All preoperative and postoperative characteristics showed a significant association with biochemical relapse. Cox regression of preoperative characteristics showed the number of positive biopsies with predominant Gleason grade 4 and/or 5 cancer to be the most accurate predictor of failure (p < 0.0001), followed by the number of positive biopsies and PSA. CART analysis distinguished between four risk groups on the basis of the same characteristics as in the Cox regression. The low-risk group consisted of 232 patients (75.1%) and the high-risk group of 17 patients (5.5%); corresponding Kaplan-Meier curves showed a 2-year PSA-free survival rate of 97% for the low-risk group and 20% for the high-risk group. Cox regression of postoperative characteristics recognized the volume of Gleason grade 4 and/or 5 as the characteristic with the strongest association with biochemical failure. CART analysis distinguished between four risk groups, using the volume of high-grade cancer as the most influential characteristic. The corresponding Kaplan-Meier curves showed for the low-risk group (n = 79; 29.6%) a PSA-free survival rate of 96% after 42 months and for the high-risk group (n = 47; 17.6%) a 21% PSA-free survival rate after 42 months. CONCLUSION: For preoperative and postoperative estimation of biochemical recurrence after RRP, a quantitative analysis of high-grade cancer, expressed by the number of preoperative biopsy cores containing high-grade cancer and the volume of cancer, proved to be the best predictor of relapse. CART analysis might be useful in advising patients for their best therapy options. However, defined characteristics of risk groups should be evaluated with new prospective data before they are used routinely. PMID- 10364653 TI - Effect of an educational multimedia prostate program on the International Prostate Symptom Score. AB - OBJECTIVE: To determine the effect of an interactive multimedia prostate education program (MMP) on self assessment of symptom scores due to benign prostatic hyperplasia (BPH). METHODS: The interactive MMP was developed including a computer-administered version of the International Prostate Symptom Score (IPSS) questionnaire. Eighty-eight men referred to the Urology Out-patients with prostatic symptoms entered the study. They first completed the IPSS on paper and secondly used the MMP before completing the computer-administered IPSS. A final feedback questionnaire enquiring into their experience including previous exposure to computers, ease of use, and value of the program content was completed. RESULTS: The use of the MMP resulted in a significant decrease in mean IPSS score from 16.6 to 13.9 (t = 7.456, d.f. = 87, p < 0.01), but no change in quality of life. Patients felt that their knowledge had increased (chi2(1) = 21.253, p < 0.01) and that they had completed the IPSS more accurately (chi2(1) = 10.227, p < 0.01) with the MMP IPSS module compared to the IPSS on paper. Previous use of patient education, patient characteristics and MMP use beyond the information required for the IPSS did not affect IPSS difference (IPSS before versus after MMP use). CONCLUSION: The use of the MMP enhanced patients' knowledge of their condition and reduced patients' IPSS score. The results were independent of previous exposure to information, previous IPSS completion, computer use and age. PMID- 10364654 TI - Complications of laparoscopic adrenalectomy in 75 patients treated by the same surgeon. AB - OBJECTIVE: We analyzed the complications of endoscopic adrenalectomy. METHODS: We retrospectively reviewed the operative and postoperative complications among 75 patients with adrenal tumors who underwent endoscopic adrenalectomy by the same surgeon. RESULTS: Five patients (6.7%) were converted to open surgery. Of these, there were 2 with metastatic adrenal carcinoma, and 1 with adrenal tuberculosis. A total of 21 patients (28%) had 24 complications (32%). There was no mortality. As for access and pneumoperitoneum-related complications, 5 cases of subcutaneous emphysema and 3 of radiating shoulder pain occurred. Intraoperative complications included 2 cases of vascular injury, 2 of organ injury, and 4 of massive bleeding (>500 ml). Postoperative complications included 2 cases of mild paralytic ileus, 2 asthma, and 1 each of angina, wound infection, retroperitoneal hematoma, and contralateral atelectasis. Except for the patients with adrenal malignancy and adrenal tuberculosis, 71% of the complications occurred among the initial 25 patients with laparoscopic adrenalectomy and 80% occurred in the initial 10 retroperitoneoscopic patients. CONCLUSION: Although endoscopic adrenalectomy is a valuable alternative to open surgery, it should be done by a skilled laparoscopist in patients with adrenal inflammatory lesions or malignancy. Careful patient selection and correct choice of surgical approach according to the tumor size and the patient's condition are the most important points for avoiding the complications of laparoscopic adrenalectomy. PMID- 10364655 TI - Primary endoscopic treatment of ureteric calculi. A review of 378 cases. AB - AIM OF THE STUDY: In the post-ESWL period, ureteroscopy represented the solution giving a second choice in the treatment of ureteral calculi in case of failure of extracorporeal lithotripsy. The aim of this study is to review a wide series of ureteral stones in which ureteroscopy combined with endoscopic lithotripsy can be chosen as the first approach for the treatment of ureteral calculi. METHODS: Between January 1994 and September 1997, 378 patients underwent ureteroscopy and endoscopic lithotripsy for ureteral stones with a miniscope associated with either a pneumatic or electropneumatic lithotriptor. Three different miniscopes were used: Olympus (8 Fr), Wolf (7 Fr) and Circon Acmi (7.7 Fr). 238 patients were male and 140 were female. The stones were localized in the upper tract of the ureter in 62 cases (16.4%), 96 (25.3%) in the mid ureter and 220 (58. 3%) in the lower ureter. RESULTS: A complete stone fragmentation with spontaneous expulsion of the fragments occurred in 354 patients (93.6%). In 22 patients (5.8%) the stones were accidentally pushed up and successfully underwent ESWL. In 38 patients (10%) the fragments were completely removed by basket. A single J polyethylene catheter was placed in 21 (5.5%) and a JJ stent in 147 patients (38. 8%). The operative time ranged from 10 to 60 min, with an average time span of 32. In 22 cases (5.8%) an iterative ureteroscopy for stenosis or incomplete fragmentation was needed. Five cases (1.3%) of ureteral perforation were successfully treated by JJ stent, and only 1 case of ureteral avulsion (upper ureter) was treated by open surgery. In the attempt of overcoming an ureteral stenosis, we had 1 case (0.2%) of ureteral reimplantation. One patient (0.2%) underwent ureterolithotomy for an extremely narrow stenosis just before the ureteropelvic junction. No relevant complication was recorded in the postoperative period. Patients were dismissed after 1- 4 days (average 1.9). Up until now, no case of postoperative ureteral stricture has been observed, although we were not able to carry out a specific follow-up in all our patients. CONCLUSIONS: Ureteroscopy with miniscopes has a high success rate (93.6%) with low morbidity and can be given as a primary approach in the management of ureteral calculi. In the lumbar ureter (especially in women) this technique can represent a good alternative to ESWL in the treatment of obstructing stones (which need stenting) or when the patient asks for a 'one-shot' treatment. PMID- 10364656 TI - Bacterial colonization of ureteral stents. AB - OBJECTIVES: The aim of this study was to assess the frequency of bacterial stent colonization and stent-associated bacteriuria, and to evaluate the significance of urinary cultures for identification of colonizing microorganisms. METHODS: A total of 93 ureteral stents from 71 patients were examined: 9 patients with permanent ureteral stenting due to malignant ureteral obstruction (27 stents), and 62 patients with temporary ureteral stents (66 stents). RESULTS: Bacteriuria and bacterial stent colonization were found in all patients with permanent stents. In patients with temporary stents, colonized stents were found in 69.3% (43/62), mainly in combination with sterile urine (45.2%, 28/62). Mean indwelling times did not differ between patients with sterile urine and sterile stents (11.8 days) and patients with sterile urine and colonized stents (11.2 days). Prophylactic antibiosis in 42/62 temporarily stented patients did not reduce colonization rates compared to patients without antibiotics (70 vs. 65%). Enterococci were the bacteria most frequently cultured from urine and stents. CONCLUSIONS: In the present study, ureteral stent colonization rates were 100% in permanently and 69.3% in temporarily stented patients. Antibiotic prophylaxis did not prevent stent colonization and should not be routinely administered. Since urinary cultures correctly identified all colonizing microorganisms in only 21%, removal/replacement and bacteriologic evaluation of ureteral stents may be necessary in case of urosepsis. PMID- 10364657 TI - Hemodynamic characterization of a functional erection. Arterial and corporeal veno-occlusive function in patients with a positive intracavernosal injection test. AB - OBJECTIVES: To characterize hemodynamically a functional/rigid erection and study the hypothesis that a positive intracavernosal injection test indicates normal arterial and corporeal veno-occlusive function. METHODS: 33 patients (mean age 39.5 +/- 9 years), who developed rigid erection during pharmacocavernosometry, included in the present study. The presence of axial rigidity was determined at steady state equilibrium intracavernosal pressure, by absence of buckling to axial force of 1 kg, applied to the erect penis and sustained for >/=15 min. Arterial and veno-occlusive hemodynamic parameters were analyzed. RESULTS: Flow to-maintain at intracavernosal pressure 150 mm Hg and mean pressure decay values ranged between 0.5-13 ml/min and 5-85 mm Hg, respectively. Flow-to-maintain values >5 ml/min were noticed in 8 patients (24. 24%), while pressure decay values >45 mm Hg in 13 patients (39.39%). Pharmacocavernosography revealed moderate opacification of venous structures in 7 cases (21.21%). Abnormal systemic-cavernosal systolic arterial pressure gradients in both cavernosal arteries were noticed in 9 patients (27.27%). All patients with flow-to-maintain values >5 ml/min had normal arterial function. CONCLUSIONS: A functional/rigid erectile response may coexist with arterial insufficiency or corporeal veno occlusive dysfunction. Presence of normal or borderline arterial inflow may compensate minimal or moderate veno-occlusive dysfunction, resulting in a functional - but not normal - erection. Such information is critical when the intracavernosal injection test is used for diagnostic purposes. PMID- 10364658 TI - Is obesity an underlying factor in erectile dysfunction? AB - OBJECTIVES: We conducted a study to evaluate the impact of obesity on erectile function in men with erectile dysfunction. METHODS: Three hundred and twenty-five consecutive patients with erectile dysfunction were evaluated. We classified the men into 2 groups according to body weight: <120% of the ideal body weight, and >/=120%. We compared several erectile capacities and the findings of penile duplex ultrasonography. RESULTS: There was a statistically significant decrease in the quality of residual erectile function in patients with obesity (penile rigidity grade 1.32 versus 1.62 in the nonobese patients). Obese patients also have an increased prevalence of vascular risk factors based on a review of the medical records and vascular impairment by duplex ultrasound study (43 and 62% in the obese patients versus 30 and 42% in the nonobese patients, respectively, p < 0.05). However, when we focused only on the patients without any vascular risk factors, no significant difference between the 2 groups was noted in the quality of residual erectile function and also the prevalence of penile vascular impairments (p > 0.05). CONCLUSIONS: These data demonstrate that obesity in itself does not seem to be an underlying factor, but does impose a risk to vasculogenic impotence by developing chronic vascular disease. PMID- 10364659 TI - Ureterovascular hydronephrosis in children: is pyeloplasty always necessary? AB - OBJECTIVE: To evaluate the efficacy of the vessel transposition technique in ureterovascular hydronephrosis in children. METHODS: Over a 25-year period, we treated 111 patients with 112 instances of ureterovascular hydronephrosis. In order to determine the obstructive effect of the vessels, we performed an intraoperative diuretic test. Using this approach, 61 patients judged to have only vascular pyeloureteral junction obstruction underwent vessel transposition. However, 50 patients in whom the intraoperative diuretic test proved doubtful needed pyeloplasty. RESULTS: Surgical success was achieved in 98% of the patients. Only 1 child treated by vessel transposition had an unsatisfactory outcome which necessitated a subsequent pyeloplasty for persistent hydronephrosis. This was due to a previously unrecognized intrinsic pyeloureteral junction obstruction. CONCLUSION: Based on our clinical experience, the intraoperative diuretic test has proven to be a safe and effective diagnostic tool in children with ureterovascular hydronephrosis. Its use may contribute to treating some cases of ureterovascular hydronephrosis without resorting to pyeloplasty. PMID- 10364660 TI - Importance of the renal resistive index in children suffering from vesicoureteral reflux. AB - OBJECTIVE: in our study, we wanted to assess changes and values of the renal resistive index (RI) in children with vesicoureteral reflux (VUR) using color Doppler ultrasound (CDUS) techniques. METHODS: We investigated 92 kidney units in 46 children (mean age 6 +/- 4 years) with VUR by means of CDUS. The RI was measured at the level of the interlobar and/or arcuate arteries using a color Doppler unit Acuson 128 XP/10. The CDUS findings were correlated with those obtained using standard renal scintigraphy. RESULTS: Children with VUR grades I III showed a RI of 0.6 +/- 0.07, whereas patients suffering from VUR grades IV and V demonstrated a significantly increased RI of 0.77 +/- 0.07 (p < 0.0001). The correlation of the CDUS measurements with the scintigraphic findings revealed a positive correlation coefficient of r2 = 0.61. CONCLUSIONS: CDUS seems to be a helpful, radiation-free, noninvasive, and inexpensive tool in children with VUR, demonstrating a significant change in the RI especially in high-grade reflux. Moreover, the RI findings represent a good correlation with standard renal scintigraphy. Therefore, CDUS with assessment of the RI allows a perfect follow up study of affected children. PMID- 10364661 TI - In utero fetal muscle biopsy: a precious aid for the prenatal diagnosis of Duchenne muscular dystrophy. AB - Prenatal diagnosis for Duchenne muscular dystrophy can usually be performed using DNA analysis. This approach would be impossible when there is only one prior affected male and no identifiable gene deletion. Therefore, in utero fetal thigh muscle biopsy with direct examination of muscle by dystrophin analysis may provide the only means of prenatal diagnosis. We report such a case in which fetal muscle biopsy was able to exclude Duchenne muscular dystrophy. A detailed literature review of the topic is provided. PMID- 10364662 TI - Antenatal therapy of Smith-Lemli-Opitz syndrome. AB - OBJECTIVES: Smith-Lemli-Opitz syndrome (SLOS) is a recessively inherited disorder caused by an inborn error of cholesterol metabolism that results in deficiency of cholesterol and accumulation of the cholesterol precursor, 7-dehydrocholesterol (DHC) and its epimer, 8-DHC. Affected patients present with congenital anomalies, growth restriction, and mental retardation. Postnatal treatment with cholesterol supplementation has been shown to improve plasma sterol levels and has resulted in improved growth and development in many patients. We hypothesized that prenatal supplementation of cholesterol could potentially arrest some of the adverse consequences of cholesterol deficiency at an earlier stage of development. METHODS: SLOS was diagnosed in the third trimester in a fetus initially identified by sonography with intrauterine growth restriction and ambiguous genitalia and confirmed by elevated levels of 7- and 8-DHC in amniotic fluid. Antenatal supplementation of cholesterol was provided by fetal intravenous and intraperitoneal transfusions of fresh frozen plasma (cholesterol level = 219 mg/dl). RESULTS: The in utero transfusions resulted in increased levels of fetal cholesterol, as measured in blood samples obtained by cordocentesis. In addition, fetal red cell mean corpuscular volume rose, which further indicated that the exogenous cholesterol was incorporated into the fetal erythrocytes. CONCLUSIONS: Antenatal treatment of SLOS by cholesterol supplementation is feasible and results in improvement in fetal plasma cholesterol levels and fetal red cell volume. SLOS may be added to the growing list of human genetic disorders for which prenatal diagnosis is available and therapeutic intervention may be possible. PMID- 10364663 TI - Induction of labor in patients with term premature rupture of membranes. Effect on perinatal outcome. AB - OBJECTIVE: To determine whether delayed induction of labor in patients with premature rupture of membranes (PROM) at term has beneficial effects on the mother or the infant. STUDY DESIGN: Retrospective analysis of our database revealed 576 patients >37 weeks of gestation with PROM, who delivered live-born infants without major congenital anomalies. We analyzed the frequencies of primary cesarean, neonatal intensive care unit (NICU) admissions, and oxytocin use by time since hospital admission and interval until onset of labor. RESULTS: NICU admission increased from 1.9% in <3 h between admission to onset of labor to 13.3% after >18 h. Admission-onset of labor interval, birth weight of <2,500 or >4,000 g and meconium were all more important determinants of NICU admission than gestational age, duration of labor, PROM, and ROM. Prolonged admission-onset of labor interval was associated with an increased risk of variable decelerations (p < 0.001). Primary cesarean rates increased progressively with longer intervals between admission and onset of labor. Stepwise discriminant function analysis revealed that labor duration, admission-onset of labor interval, gestational age, and birth weight of <2,500 g were all more important determinants of primary cesarean delivery than the durations of PROM or ROM. CONCLUSIONS: The increased frequencies of NICU admission, variable decelerations, and primary cesarean suggest that delayed labor induction after hospital admission was linked to worsened perinatal outcomes. These results may have been influenced by usually performing a single digital examination as part of initial evaluation of term patients who present with PROM. Based on our data, we suggest immediate induction for PROM at term, especially if digital examination has been performed. PMID- 10364664 TI - Can we suspect fetal down syndrome by heart evaluation during the second half of pregnancy? AB - Thirty fetuses with Down syndrome, who had detailed fetal echocardiography and sonography at the tertiary center with videotape recordings, were retrospective analyzed by one observer with a specially prepared flow sheet. The mean gestational age of the fetuses at the time of the study was 31 +/- 5.6 (minimum 21, maximum 39) weeks. The 'main' fetal abnormalities were congenital heart defect (CHD): in 13 cases (43.3%) an abnormal 4-chamber view was recorded, including 6 cases (20%) of isolated CHD and 7 (23.3%) of coexisting CHD + extracardiac malformation. Of the 13 cases of CHD, there were 12 cases of atrio ventricular canal and 1 case of ventricular septal defect. Normal heart anatomy was recorded in 17 cases (56.7%), including 2 with tricuspid value regurgitation. From the videotape recordings also some 'minor' abnormalities were noticed in a few cases such as: femur length shortening; sandal gap; pericardial effusion; macroglossia; echogenic bowel; absent diastolic flow in the umbilical artery, and others. The prevalence of CHD in the study group was similar to the prevalence of CHD in the comparison group of 20 newborns with Down syndrome, born during the same period of time at the same institution, who had not had prenatal scanning at all (chi2, p > 0.05). CONCLUSIONS: (1) the main 'major' abnormality which might be detected in a fetus with Down syndrome after 20 weeks of pregnancy is CHD, which was presented in 43.3% of this series; (2) the presence of any extracardiac malformation should prompt the sonographer for detailed heart evaluation, and (3) fetal echocardiography may increase the accuracy of 'genetic sonogram' in Down syndrome. PMID- 10364665 TI - Prenatal diagnosis of Down's syndrome in the presence of isolated Ebstein's anomaly. AB - Ebstein's anomaly is a rare congenital cardiac defect, characterized by the displacement of the tricuspid valve into the right ventricle, that occurs approximately once in 20,000 live births. The association of Ebstein's anomaly and chromosomal abnormalities, such as Down's syndrome, is extremely unusual. Prenatal diagnosis of trisomy 21 in a fetus with isolated Ebstein's anomaly has not been previously reported. PMID- 10364666 TI - Effect of amnioinfusion on the outcome of prenatally diagnosed gastroschisis. AB - OBJECTIVE: Following recent data showing that an inflammatory response exists in the amniotic fluid of gastroschisis-affected fetuses, we hypothesized that amniotic fluid exchange or amnioinfusion would improve the prognosis of prenatally diagnosed gastroschisis. METHODS: We compared the outcome of prenatally amnioinfused fetuses with gastroschisis to non-amnioinfused fetuses with gastroschisis. 10 patients undergoing this procedure were matched with 10 patients of our previous study. Comparisons were done on data including surgical procedure, follow-up in the NICU and the gastro-pediatric unit. RESULTS: Our results show that gastroschisis-affected fetuses undergoing amnioinfusion had a lower duration of curarization after surgery (2.2 +/- 1.9 vs. 6.8 +/- 6.9 days, p = 0.019), a shorter delay before full oral feeding (49.7 +/- 21.5 vs. 72.3 +/- 56.6 days, NS), and a shorter overall length of hospitalization (59.5 +/- 19.7 vs. 88.5 +/- 73.6 days, NS). We confirmed our previous data showing that amniotic fluid displays a chronic inflammation profile. CONCLUSION: Our data suggest that amnioinfusion could improve the outcome of gastroschisis affected fetuses. The hypothesis by which this improvement could be due to a reduction of an inflammatory response remains to be proved. PMID- 10364667 TI - Comparison of unstimulated and stimulated behaviour in human fetuses with congenital abnormalities. AB - Unstimulated (passive) and stimulated behaviour [fetal heart rate (FHR) and movements (FA)] was studied in 32 normal fetuses and 10 fetuses with congenital abnormalities (CA). FHR and FA were recorded using a single 1.5-MHz ultrasound transducer and analysed by computer. A 5-second vibroacoustic stimulus (VAS) ('electronic artificial larynx') was used for the stimulation studies. Thus, passive and stimulated behaviour could be studied in a group of fetuses with known pathologies. One hour was used as the recording time for the passive studies and 20 min for the stimulation studies (10 min pre- and 10 min post-VAS). All CA fetuses had abnormalities of FHR and/or FA on recording passive behaviour compared to normal fetuses. However, 4 of the 10 fetuses with CA had responses to VAS that were within the normal range for both FHR and FA. We do not feel that computerised assessment of stimulated behaviour in fetuses with CA confers any advantage over analysis of passive behaviour. PMID- 10364668 TI - Three-dimensional imaging of the postmortem fetus by MRI: early experience. AB - OBJECTIVE: The feasibility and significance of three-dimensional (3D) visualization of the postmortem fetus using magnetic resonance imaging (MRI) was investigated. METHODS: 3D reconstruction of sectional MRI data sets from 8 postmortem fetuses was performed. RESULTS: Fetal configurations and internal structures, both normal and pathological, were clearly demonstrated by 3D display. CONCLUSION: This new technique provides high quality fetal 3D images for postmortem morphological diagnosis and interactive visual teaching. It may eventually have applications in prenatal diagnosis and the preoperative simulation of fetal surgery. PMID- 10364669 TI - Amnioinfusion in the evaluation of fetal obstructive uropathy: the effect of antibiotic prophylaxis on complication rates. AB - OBJECTIVE: Amnioinfusion plays an important role in the intrauterine evaluation and treatment of fetal obstructive uropathy. However, it may significantly increase the risk for chorioamnionitis, premature rupture of membranes and premature labor. We evaluated the impact of prophylactic antibiotics on postamnioinfusion complications. METHODS: Thirty pregnancies complicated by fetal obstructive uropathy, treated by amnioinfusion and with documentation of pregnancy outcome were identified from our database. Pregnancy outcomes were compared between patients who were treated with prophylactic antibiotics and those with no prophylaxis. RESULTS: Chorioamnionitis was diagnosed in 3 out of 15 (20%) patients who did not receive prophylactic antibiotics as compared to 2 (13%) in the treated group. The overall rate of serious obstetrical complication was significantly higher in the untreated group (66 vs. 20%; p = 0. 021). Patients receiving prophylactic antibiotics delivered at a significantly greater gestational age than those who did not receive antibiotics (34.0 +/- 3.7 vs. 31.3 +/- 1.9 weeks, respectively; p = 0.018). CONCLUSIONS: Our study supports the use of oral prophylactic antibiotics as being effective in reducing the previously observed significant risks associated with amnioinfusion in fetal obstructive uropathy. PMID- 10364670 TI - Evaluation of fetal echogenic bowel in the second trimester. AB - Previous studies cite different possible etiologies for fetal echogenic bowel (FEB). The purpose of this study was to evaluate the possible etiologies for second-trimester FEB, and to provide clinical guidelines for evaluation of this finding. The study included 79 patients diagnosed with FEB in the second trimester. Fifteen cases (19%) were associated with maternal vaginal bleeding. Of these, 12 patients underwent amniocentesis, 9 of which had visible blood products in the amniotic fluid. Seven cases (8.9%) had associated severe malformation. Seven other cases (8.9%) were noted in multifetal pregnancies. Five fetuses (6.3%) had evidence of bowel obstruction or perforation not associated with cystic fibrosis (CF). Chromosomal aberrations were found in 5 fetuses (6.3%). Intrauterine infection with cytomegalovirus, herpes simplex virus, varicella zoster virus, or parvovirus B-19 was documented in 5 patients (6.3%). Three cases (3.8%) were associated with subsequent unexplained stillbirth. Two fetuses (2.5%) were found to be affected by CF. Finally, in 30 cases (38%), no obvious reason for FEB was found. We conclude that the evaluation of second-trimester FEB should include targeted ultrasound for associated malformations, infectious studies, DNA analysis for CF mutations, amniocentesis for chromosomal analysis and evaluation of the amniotic fluid for degraded blood products, and an autopsy in cases of stillbirth. Even when no apparent reason is found, pregnancies should be considered at high risk for poor outcome. PMID- 10364671 TI - Differential effect of advanced maternal age on prenatal diagnosis of trisomies 13, 18 and 21. AB - Nondisjunction associated with advanced maternal age, a well-established factor in the etiology of autosomal trisomy, should equally affect all chromosomes. In this study we evaluate the association of advanced maternal age with the occurrence of potentially viable autosomal trisomies (13, 18 and 21). 275 aneuploid pregnancies were ascertained prenatally and were grouped according to chromosome anomaly diagnosed. Mean maternal age was significantly younger (p = 0.009) in pregnancies affected by trisomy 13 than in pregnancies with trisomy 21. An intermediate mean maternal age was observed in pregnancies affected by trisomy 18. Our study shows a trend of the more severe, but potentially viable, autosomal trisomies to be diagnosed at younger maternal age. This may substantiate the 'relaxed selection hypothesis' proposed to explain the association of aneuploid conceptions with advanced maternal age. PMID- 10364672 TI - Distribution of neural tube defects as a function of maternal weight: no apparent correlation. AB - OBJECTIVES: Maternal nutritional deficiency is an important predisposing factor to congenital neural tube defects (NTDs). It was hypothesized that obese women may have an increased risk for NTDs. The aim of the present study was to address this question in a large cohort. METHODS: A total of 72,915 consecutive cases of biochemical screening that had documented maternal weights and pregnancy outcomes were identified from the Quest Diagnostic Laboratories database. Patients were divided into five ranges of maternal weights, and the incidence of NTDs was calculated for each group. Based on the different definitions of maternal overweight, the data were also analyzed based on 2 groups only, obese and nonobese, using three cutoff points. RESULTS: Seventy-nine pregnancies were complicated by NTDs (incidence of 1.08 per 1,000 pregnancies). Differences between maternal weights ranges were not found to be statistically significant (chi2 = 5.997, p = 0.19, power = 0.99). Differences between obese and nonobese mothers were not found to be statistically significant for all three analyses as well. CONCLUSIONS: Our present results do not support an association between maternal obesity and NTDs. PMID- 10364673 TI - Maternal fertility is not affected by fetal surgery. AB - The purpose of this report is to assess the impact of fetal surgery on future maternal fertility, subsequent pregnancy outcome, and the incidence of pregnancy complications. Retrospective data were collected on 70 mothers who underwent fetal surgery between April 1981 and June 1996. Indications for open hysterotomy fetal surgery included congenital diaphragmatic hernia (n = 44), congenital cystic adenomatoid malformation of the lung (n = 11), urinary obstruction (n = 9), sacrococcygeal teratoma (n = 4), heart block (n = 1), and acardiac-acephalic twin reduction (n = 1). The following data were obtained: number of pregnancy attempts, number of successful pregnancies, pregnancy outcome including obstetrical and neonatal complications, and infertility after fetal surgery. There were 45 respondents, of whom 35 attempted subsequent pregnancies. Thirty two were successful, resulting in 31 livebirths. Two women had a strong prefetal surgery history of infertility, 1 has only attempted to conceive for 3 months. We report this experience because the effect of open fetal surgery on futrue fertility is such an important question for our patients and referring physicians. This analysis suggests that hysterotomy and open fetal surgery has a negligible impact on maternal fertility. PMID- 10364674 TI - Screening for mutations in the promoter and the coding region of the IGFBP1 and IGFBP3 genes in Silver-Russell syndrome patients. AB - In the present study we sought to identify genetic variation in genes for insulin like growth factor binding proteins 1 and 3 (IGFBP1, IGFBP3) in 7p12-13 which through alteration of protein function or level of expression might contribute to the manifestation of Silver-Russell syndrome. Genomic DNA samples from 49 Silver Russell syndrome (SRS) patients and from unaffected controls were investigated by single-strand conformation analysis. Overlapping polymerase chain reaction fragments covered the whole coding sequences as well as the 5' untranslated region of the IGFBP1 and IGFBP3 genes. We detected 3 new polymorphisms in the transcribed sequence of IGFBP1, one amino acid polymorphism in exon 1 of IGFBP3 and four variants in its promotor region and in intron 1. They all occurred in similar frequencies in SRS patients and in controls. Thus, paternally inherited mutations in the promoter and coding regions of IGFBP1 and IGFBP3 genes play neither a major nor a minor role in the etiology of SRS. The newly detected polymorphisms in the coding region are powerful tools for analysis of imprinting status and for detection of possible changes in the imprinting patterns of the two genes. PMID- 10364675 TI - Localization of the gene responsible for familial benign polycythemia to chromosome 11q23. AB - Familial benign polycythemia (FBP) (OMIM 263400) is a rare autosomal recessive condition characterized by erythrocytosis, normal leukocyte and platelet counts, normal uric acid level, and usually increased erythropoietin production. There is a high incidence of this disorder in Chuvashia (Russian Federation), probably due to a founder effect. In an attempt to locate the gene responsible for this disorder, we have carried out linkage studies in 12 Chuvash families, with 35 affected and 32 unaffected members. Linkage to the erythropoietin and erythropoietin receptor loci was excluded, and the FBP gene was assigned to the region of chromosome 11q23 between D11S4142 and D11S1356, with a maximal lod score of 6.61. PMID- 10364678 TI - Analysis of estrogen receptor dinucleotide polymorphism by capillary gel electrophoresis with a population genetic study in 180 Finns. AB - We developed a suitable method for analysing dinucleotide repeats found in the upstream of human alpha-estrogen receptor (ER) gene by applying capillary electrophoresis and automatic analysis. This method omits the gel-casting step as well as difficult handling of long polyacrylamide sequencing gels. Use of radioactive materials is also avoided. Using this method, the frequency distribution of ER alleles, determined in 180 Finnish individuals showed two peaks at 12 and 14 repeats (166 and 168 bp) and also at 22 and 24 repeats (184 and 186 bp). The overall distribution of alleles seemed to be similar to that found among Italian and Japanese populations. PMID- 10364676 TI - Detection of the most common G6PD gene mutations in Chinese using amplification refractory mutation system. AB - Glucose-6-phosphate dehydrogenase (G6PD) is the most common human enzymopathy. To date more than 122 mutations in the G6PD gene have been discovered, among which 12 point mutations are found in the Chinese. The 2 most common mutations, G1388A and G1376T, account for more than 50% of mutations representing various regions and ethnic groups in China. Setting up a simple and accurate method for detecting these mutations is not only useful for studying the frequency of the G6PD genotypes, but also for finding new mutations. The purpose of this study was to find a simple, inexpensive and accurate method for detecting these common mutations. The amplification refractory mutation system (ARMS) method was used in this study. Samples from 28 G6PD-deficient males were investigated. The natural and mismatched amplification and restriction enzyme digestion method was used as a standard method to evaluate the nature of the point mutations. Sixteen cases were found carrying the G1388A mutation and 12 the G1376T mutation. Fourteen cases of G1388A and 10 cases of G1376T were confirmed by ARMS. Four cases were not in concordance with the results obtained by the mismatched amplification restriction enzyme digestion. These 4 cases were then judged by direct PCR sequencing at exon 12. The DNA sequencing data supported the results obtained by ARMS. Thus we concluded that the ARMS is a rapid, simple, inexpensive and accurate method for detecting the most common G6PD gene mutations among the Chinese. PMID- 10364679 TI - Testing for linkage under robust genetic models. AB - Robust genetic models are used to assess linkage between a quantitative trait and genetic variation at a specific locus using allele-sharing data. Little is known about the relative performance of different possible significance tests under these models. Under the robust variance components model approach there are several alternatives: standard Wald and likelihood ratio tests, a quasilikelihood Wald test, and a Monte Carlo test. This paper reports on the relative performance (significance level and power) of the robust sibling pair test and the different alternatives under the robust variance components model. Simulations show that (1) for a fixed sample size of nuclear families, the variance components model approach is more powerful than the robust sibling pair approach; (2) when the number of nuclear families is at least approximately 100 and heritability at the trait locus is moderate to high (>0.20) all tests based on the variance components model are equally effective; (3) when the number of nuclear families is less than approximately 100 or heritability at the trait locus is low (<0. 20), on balance, the Monte Carlo test provides the best power and is the most valid. The different testing procedures are applied to determine which are able to detect the known association between low density lipoprotein cholesterol and the common genotypes at the locus encoding apolipoprotein E. Results from this application show that the robust sibling pair method may be more effective in practice than that indicated by simulations. PMID- 10364677 TI - Simple tandem repeat polymorphisms in the neuronal nitric oxide synthase gene in different ethnic populations. AB - Allelic frequencies of a CA dinucleotide repeat in exon 29 and an intronic AAT trinucleotide repeat in the neuronal nitric oxide synthase (NOS1) gene were determined by simple sequence length polymorphism (SSLP) in 305 American Caucasian and 105 African-American healthy subjects. There were highly significant differences in allele frequencies between the two ethnically diverse study populations. PMID- 10364680 TI - The LWb blood group as a marker of prehistoric Baltic migrations and admixture. AB - Archaeological findings and historical records indicate frequent migrations and exchange of genetic material between populations in the Baltic Sea area. However, there have so far been very few attempts to trace migrations in this area using genetic markers. We have studied the Baltic populations with respect to exceptional variations in the frequencies of the Landsteiner-Wiener (LW) blood group. The frequency of the uncommon LWb gene was high in the Balts, around 6% among Latvians and Lithuanians, very low among the other western Europeans (0 0.1%) and apparently absent in Asiatic and African populations. From the Baltic region of peak frequency there was a regular decline of LWb incidence (a descending cline) in the neighboring populations: 4.0% in the Estonians, 2.9% in the Finns, 2. 2% in the Vologda Russians, and 2.0% in the Poles. Thus the distribution of LWb suggests considerable and extensive Baltic admixture, especially in the north and northeast direction. In Southern Sweden with an LWb frequency of 0.3%, the Baltic influence appeared slight, while in the population of the Swedish island Gotland in the middle of the Baltic Sea there was a significantly increased LWb frequency of 1.0% compared with that of Western European countries. The distinction of codominantly inherited LW antigenic forms, LWa and LWb (previously Nea), is known to be due to a single base substitution. Based on our population data, it is plausible that the expansion of this point mutation occurred only once during human history. Furthermore, our data indicate that the expansion of the LWb mutation occurred in Balts and that LWb can be considered a 'Baltic tribal marker', its presence in other populations being an indicator of the degree of Baltic genetic influence. PMID- 10364681 TI - Familiality of quantitative metabolic traits in Finnish families with non-insulin dependent diabetes mellitus. Finland-United States Investigation of NIDDM Genetics (FUSION) Study investigators. AB - Type 2 diabetes mellitus (NIDDM) is a complex disorder encompassing multiple metabolic defects. There exists strong evidence for a genetic component to NIDDM; however, to date there have been few reports of linkage between genetic markers along the genome and NIDDM or NIDDM-related quantitative traits. We sought to determine whether individual quantitative traits which determine glucose tolerance exhibit familiality in Finnish families with at least one NIDDM affected sibling pair. Tolbutamide-modified frequently sampled intravenous glucose tolerance tests (FSIGT) were performed on unaffected offspring (n = 431) and spouses (n = 154) of affected sibling pairs sampled for the Finland-United States Investigation of NIDDM Genetics (FUSION) study. FSIGT data were analyzed using the Minimal Model to obtain quantitative measures of insulin sensitivity (SI), glucose effectiveness (SG), and insulin secretion assessed as the acute insulin response to glucose (AIR). The disposition index (DI), a measure of insulin resistance-corrected beta-cell function, was also derived as the product of SI and AIR. Variance components analysis was used to determine for each trait, the heritability (h2), the proportion of the total trait variance accounted for by additive genes. After adjustment for age, gender, and body mass index, h2 estimates were: SG: 18 +/- 9%, SI: 28 +/- 8%, AIR: 35 +/- 8%, and DI: 23 +/- 8%. We conclude that there is strong evidence for modest heritability of Minimal Model-derived NIDDM-related quantitative traits in unaffected spouses and offspring of Finnish affected sibling pairs. PMID- 10364682 TI - Mutation analysis in patients with congenital adrenal hyperplasia in the Spanish population: identification of putative novel steroid 21-hydroxylase deficiency alleles associated with the classic form of the disease. AB - Steroid 21-hydroxylase deficiency, due to the genetic impairment of the CYP21 gene, is a major cause of congenital adrenal hyperplasia (CAH). In about 80% of the cases, the defect is related with the transfer of deleterious point mutations from the CYP21P pseudogene to the active CYP21 gene. Sixteen different point mutations have been searched for in 60 Spanish patients with the classic form of CAH and 171 unaffected family members, using selective amplification of the CYP21 gene followed by allele-specific oligonucleotide hybridization (PCR-ASOH) and sequencing analysis. While 31.9% of the disease alleles carry CYP21 deletions or large gene conversions, around 58% of the alleles carry single point mutations. Corresponding segregation of mutations was found in every case indicating that none of them has apparently appeared de novo. The most frequent mutations found in our sample are i2G, V281L, R356W, Q318X, P453S and F306+t, with rates of 30, 14.2, 10, 9.2, 9.2 and 7. 5%, respectively. We found similar frequencies for the A and C polymorphism at position 656 (40 and 31.5%, respectively) in wild-type alleles for the i2G mutation. Around 10% of the alleles, for which no mutations were identified by searching for the sixteen previously known mutations, are currently being sequenced and new possible mutations and polymorphisms have been identified. PMID- 10364683 TI - A novel G to A substitution at nucleotide 1734 of the FBN1 gene predicting a C534Y mutation responsible for marfan syndrome. PMID- 10364684 TI - Gln --> Arg 191 polymorphism of paraoxonase and Parkinson's disease. AB - We investigated the Gln --> Arg 191 polymorphism in paraoxonase (PON1) in St. Petersburg population, in three clinically differentiated groups of patients with Parkinson's disease (PD) and in the symptomatic tremor group. A new approach for Gln --> Arg 191 PON1 polymorphism genotyping is suggested. No significant differences in the groups studies as compared to the controls was observed. PMID- 10364685 TI - New AccI polymorphism in the follicle-stimulating hormone beta-subunit gene and its prevalence in three Southeast Asian populations. AB - A thymine-cytosine substitution was identified in exon 3 (codon 76, TAT to TAC) of the human follicle-stimulating hormone (FSH) beta-subunit gene. The nucleotide change led to creation of an AccI digestion site. The frequencies of the A allele (with AccI site) in Chinese (n = 201), Malays (n = 168) and Indians (n = 132) were 0.358, 0.333 and 0.402, respectively. The new FSH beta-subunit marker may be useful in gene tracking and association studies. PMID- 10364686 TI - Invasion strategies of the oral pathogen porphyromonas gingivalis: implications for cardiovascular disease. AB - Microorganisms have evolved a variety of mechanisms designed to evade detection and/or destruction by the host. Many pathogens evade host defenses by invading cells, thus providing the bacterium with an environment free of competing microorganisms. Adherence and invasion are active processes in which microorganisms often use host proteins and enzymes to gain entry into the cell, thus stimulating their own uptake. The investigation of invasion by the periopathogen Porphyromonas gingivalis is in its infancy in comparison with that of the enteric pathogens. However, recent studies with P. gingivalis have revealed that these organisms have developed invasion strategies and mechanisms similar to those of the enteric pathogens for both epithelial and endothelial cells. The study and elucidation of the mechanisms by which microorganisms such as P. gingivalis persist in chronic infection will provide valuable insight into the pathogenesis of P. gingivalis-mediated periodontal disease. The ability to multiply in and to activate endothelial cells may be one of the pathogenic mechanisms exerted by P. gingivalis that may explain the recently described association between this organism and cardiovascular disease. PMID- 10364688 TI - Accumulation of genetic alterations in brain metastases of sporadic breast carcinomas is associated with reduced survival after metastasis. AB - Tumor progression is characterized by stepwise accumulation of genetic alterations. To identify alterations associated with breast cancer metastasis, an analysis of comparative loss of heterozygosity (LOH) was performed on 38 primary sporadic breast carcinomas and 16 distant metastases. Two loci at 5q21 and 18q21 were chosen because of their reported increased deletion frequency in metastatic tumors. LOH at 17q21, 13q12-13, 17p13.1 and 11q22-23 was analyzed to determine whether there is a specific involvement of these breast cancer-associated gene loci in the metastatic process. Our data show that distant metastases are characterized by markedly increased LOH frequency at all loci examined. In both gene locus groups, significantly more distant metastases are affected by combined LOH. Furthermore, a significantly reduced postmetastatic survival time has been observed in patients with brain metastases affected by synchronous allelic loss at the four breast cancer-associated gene loci. Our results suggest that cumulative LOH of breast cancer-related gene loci is associated with a more aggressive phenotype of metastatic breast tumors. PMID- 10364687 TI - Transepithelial migration of neutrophils. AB - Neutrophils are a key cell type of nonadaptive immune system and are the first phagocytic cell type that reaches mucosal inflammatory sites. On the last stage of their journey from the blood stream to a mucosal surface, neutrophils cross a generally sealed epithelium by migrating along the paracellular pathway to the luminal side of the epithelial layer. This last step involves a specific receptor mediated adhesion event of the neutrophil to the epithelium, followed by a rapid and highly coordinated reversible opening of the epithelial intercellular junctions that allows the transmigration of the neutrophils. Although we do not yet understand the molecular mechanisms that mediate this transmigration process, the last years witnessed the discovery of the first neutrophil and epithelial cell surface proteins critically involved in transepithelial migration of neutrophils. PMID- 10364689 TI - Tissue-specific posttranscriptional downregulation of expression of the S100A4(mts1) gene in transgenic animals. AB - The S100A4(mts1) is a gene associated with generation of metastatic disease. In order to analyze the consequences of alteration of the pattern of expression of the S100A4(mts1) gene we obtained strains of transgenic mice bearing the S100A4(mts1) gene under the control of a ubiquitous and constitutive 3-hydroxy-3 methylglutaryl CoA reductase (HMGCR) gene promoter. In transgenic animals the expression of the transgene RNA was detected in all organs, but only some of the organs showed elevated levels of the protein. Expression of the S100A4(Mts1) protein was downregulated in the organs that normally do not express the gene in the wild-type animal. The transgene RNA is detected in the polysomes indicating that it could be translated into the S100A4(Mts1) protein. The specificity of the S100A4(Mts1) protein expression is determined by a complex mechanism including regulation of translation and/or posttranslational degradation. PMID- 10364690 TI - Stimulation of motility of human renal cell carcinoma by SPARC/Osteonectin/BM-40 associated with type IV collagen. AB - SPARC is known to be important in development and tissue remodelling. Here, we examined the effects of SPARC (secreted protein, acidic and rich in cysteine; osteonectin) derived from a rat osteosarcoma cell line on migration of renal cell carcinoma (RCC) by a Boyden chamber assay. YCR RCC cells migrated through type IV collagen-coated filters without stimuli (basal level). SPARC in the lower compartment stimulated chemotactic activity to 120% of the basal level, whereas premixing of YCR with purified SPARC before inoculation reduced their migration to 72% of the basal level. Furthermore, SPARC mixed with type IV collagen more efficiently stimulated their migration in a concentration-dependent manner (up to 170% of the basal level). This suggests that SPARC bound to type IV collagen plays a role in tumor invasion. PMID- 10364691 TI - Characterization of ionotropic glutamate receptors in rat hypothalamus, pituitary and immortalized gonadotropin-releasing hormone (GnRH) neurons (GT1-7 cells). AB - Evidence from various sources suggested that the Gonadotropin-Releasing Hormone (GnRH) neuron does not contain glutamate receptors. Northern analysis of the hypothalamus showed the presence of NMDAR1, GluR1, GluR4 and GluR6 mRNA, while the pituitary showed the presence of NMDAR1, GluR1 and GluR6 mRNA. Western blot analysis also showed the presence of NMDAR1 and GluR1 protein. Since there are relatively few GnRH neurons in the hypothalamus, and GT1-7 cells have been considered to be a GnRH neuronal cell line, GT1-7 cells were studied in detail. GT1-7 cells contained NMDAR1 mRNA levels as shown by Northern analysis but did not contain GluR1, GluR4, or GluR6 mRNA. They did not show the presence of NMDAR1 and GluR1 protein by Western analysis. In addition, GT1-7 cells showed no NMDA receptor binding using the competitive inhibitor CGP-39563 and the noncompetitive inhibitor MK-801. Likewise, no binding was detected for kainate receptors. However, a small amount of binding for AMPA receptors was found in GT1-7 cells. GT1-7 cells did not exhibit glutamate toxicity and NMDA failed to elicit inward currents using patch-clamp techniques, although GABA did induce currents in the cells. As a whole, these studies suggest that GT1-7 cells lack or possess only low levels of ionotropic glutamate receptors. PMID- 10364692 TI - Regulation of gonadal and somatotropic axis by chronic intraventricular infusion of insulin-like growth factor 1 antibody at the initiation of puberty in male rats. AB - There has been increasing experimental evidence to suggest that insulin-like growth factor 1 (IGF-I) may be one of the essential regulators in the reproductive system of the rat. IGF-I is synthesized in the hypothalamus and IGF I immunoreactivity increases during puberty. Consequently we hypothesized that centrally located IGF-I might contribute to the initiation of puberty. Centrally located IGF-I was immunoneutralized to assess this hypothesis. Male Wistar rats, 28 days old, were infused intracerebroventricularly with specific purified IgGs from rabbit IGF-I antiserum (IGF-I-Ab). The intracerebroventricular administration of IGF-I-Ab resulted in a reduction in testicular weight and consequently in delayed pubertal development. There was also a reduction in serum testosterone, pituitary immunoreactive (IR) luteinizing hormone (LH) and serum IR follicle-stimulating hormone (FSH). The accumulation of betaLH mRNA was not modified, whereas betaFSH mRNA was increased. An increment in the serum growth hormone (GH) levels was also observed. There were no significant alterations in hypothalamic IR growth hormone releasing factor content, although IR somatostatin (SRIH) content was increased by IGF-I-Ab. The body weight gain remained unaltered. As a whole, our study suggests that centrally located IGF-I influences pubertal development, production and release of gonadotropins and supports the finding that endogenous centrally located IGF-I plays a role at the initiation of puberty in the male rat. It also gives support to the physiological role of centrally located IGF-I in the release of GH mediated by hypothalamic SRIH at the initiation of puberty. PMID- 10364693 TI - Cyclical variations in the abundance of leptin receptors, but not in circulating leptin, correlate with NPY expression during the oestrous cycle. AB - We have demonstrated previously that pharmacological doses of oestradiol decreased leptin receptor expression in the hypothalamus. We therefore analysed leptin receptor expression during the oestrous cycle in the rat, to establish if acute changes in oestradiol affect leptin receptor expression under physiological conditions. Radioactive in situ hybridization histochemistry was used to measure the gene expression under investigation. Total leptin receptor transcript levels were lowest in pro-oestrus in the choroid plexus, these changes correspond inversely with levels of circulating oestradiol in the rat 4-day oestrous cycle. In contrast full-length leptin receptor levels in both arcuate and ventromedial nuclei did not correspond to the levels of total leptin receptor in the same areas of the hypothalamus or serum levels of oestradiol. Full-length leptin receptor expression in the arcuate nucleus was negatively correlated with neuropeptide Y (NPY) expression (r = 0.447, p < 0. 05) in the same nucleus. Arcuate nucleus NPY expression did not correlate significantly with the expression of total leptin receptors in the arcuate nucleus (r = 0.080) or serum leptin levels (r = 0.251). Our results demonstrate that leptin receptor expression is regulated during the oestrous cycle. The unchanged serum leptin levels during the oestrous cycle together with the correlation between the expression of leptin-RL and NPY provide circumstantial evidence that regulation of leptin receptor abundance in the hypothalamus governs the biological actions of leptin. PMID- 10364694 TI - Estrogenic stimulation of hypothalamic-limbic system metabolism in ageing diabetic C57BL/KsJ mice. AB - The therapeutic influences of estrogen treatment on age- and diabetes-related declines in regional brain glucose utilization (RBGU) rates were evaluated in 8- to 20-week-old female C57BL/KsJ normal (+/?) and diabetic (db/db) mice. Following either oil vehicle (oil: 0.1 ml) or estradiol (E: 1 microgram/3.5 days) treatments starting at 3 weeks of age, RBGU rates were subsequently determined at 8, 12, 16 and 20 weeks of age. A gradual decline in the basal rate of brain glucose utilization was observed in all control (oil- and E-treated) groups between 8 and 20 weeks. Expression of the hyperglycemic-obese diabetes syndrome in db/db mice resulted in a significant reduction in RBGU rates between 8 and 20 weeks relative to control values. In estrogen-sensitive hypothalamic, septal and amygdaloid regions, E therapy modulated RBGU rates in db/db mice relative to oil treated diabetics, but did not significantly alter utilization rates in +/? mice. In cortical samples, E therapy had no significant influence on glucose utilization rates in either control or diabetic groups. A noticeable pattern of maturation-associated decline in CNS glucose utilization rates in all brain regions resulted in comparable regional metabolic indices being exhibited by all groups at 20 weeks of age, with the exception of the diabetes-associated exacerbation of RBGU rates in the oil-treated db/db group. These data demonstrate that the normal development-related decline in regional brain carbohydrate metabolism is accelerated by the diabetes syndrome, and that E therapy can modulate the syndrome-associated suppression of glucose utilization in steroid sensitive CNS loci. These data suggest that the depressive influences of the diabetes syndrome on brain carbohydrate utilization rates may be therapeutically modified in recognized CNS regions possessing steroid-sequestering, metabolically responsive neurons. PMID- 10364695 TI - Changes in hypothalamic estrogen receptor-containing cell numbers in response to feed restriction in the female lamb. AB - The mechanism whereby undernutrition enhances the ability of estradiol (E) to inhibit reproductive activity is unknown. This study aimed to determine the effect of feed restriction on E receptor (ER)-containing cell numbers in the female sheep hypothalamus. Ovariectomized lambs at 7 months of age received either ad libitum (AL; n = 5) or restricted (FR; n = 10) levels of feed intake. Lambs were weighted weekly and FR lambs fed to lose approximately 15% of their initial body weights over 7 weeks, at the end of which jugular blood samples were collected at 10-min intervals for 5 h to assess the patterns of LH release. After blood collection, lambs were euthanized and hypothalami collected for immunocytochemical detection of ER. Based on LH secretory profiles, FR lambs were subdivided into two groups. The first group (FR + LH; n = 5) exhibited patterns of LH release similar to AL controls. LH secretion in the second group (FR-LH; n = 5) was obviously suppressed. Numbers of ER-containing cells did not differ significantly (p > 0.10) among treatment groups in the bed nucleus stria terminalis, anterior hypothalamic area and arcuate nucleus. ER-containing cell numbers were greater (p < 0.05) in the preoptic area (POA) but less (p < 0.05) in the ventromedial/ventrolateral hypothalamus (VMH/VLH) for FR-LH lambs compared to AL animals. Notably, for both the POA and VMH/VLH, ER-containing cell numbers in the FR + LH animals were intermediate and did not differ (p > 0.10) from either FR-LH or AL lambs. These results suggest that feed restriction differentially alters ER-containing cell numbers in specific regions of the ovine hypothalamus (numbers increased in the POA but decreased in the VMH/VLH). These changes may, at least in part, represent a mechanism whereby undernutrition enhances the ability of E to inhibit reproduction. PMID- 10364696 TI - GABAergic regulation of lordosis: influence of gonadal hormones on turnover of GABA and interaction of GABA with 5-HT. AB - The role of GABAergic neurons in activating female sexual behavior and possible mechanisms for GABAergic effects on behavior were examined in female rats. First, effects of the ovarian hormones estrogen and progesterone (P), at doses which promote lordosis, on levels and turnover/activity of GABA, were examined in brain areas which regulate lordosis. Utilizing AOAA, an inhibitor of GABA degradation, the accumulation rate of GABA (turnover/activity) was assessed in ovariectomized (Ovx), Ovx + estrogen and Ovx + estrogen + P-treated rats. Estradiol increased GABA accumulation rates in the arcuate-median eminence and in the area dorsal to and surrounding the VMN (VMN-S). P administration following estrogen priming enhanced GABA turnover in the medial preoptic area (mPOA) and further increased turnover in the VMN-S while GABA turnover decreased in the dorsomedial nucleus. No effects of hormones were noted in the VMN itself or in the dorsal midbrain central gray. Reverse dialysis of the GABAA antagonist bicuculline into the basomedial hypothalamus was associated with a time-dependent inhibition of lordosis and a 300% increase in 5-HT release in the basomedial hypothalamus as measured by in vivo dialysis. These results provide additional evidence that GABAergic neurons mediate the physiological regulation of female sexual behavior and suggest that such mediation may involve an interaction with 5-HT containing neurons. PMID- 10364697 TI - Effect of direct application of estrogen aimed at lateral septum or dorsal raphe nucleus on lordosis behavior: regional and sexual differences in rats. AB - The role of estrogen in lordosis-inhibiting systems in the lateral septum or the dorsal raphe nucleus was investigated in female and male rats. Ovariectomized rats received implantation of 22-gauge guide cannulae to the bilateral or right side of the lateral septum (LS and rLS, respectively), the dorsal raphe nucleus (DRN) or bilateral cortex (CX). In castrated male rats, bilateral implantations of the cannulae to the LS were carried out (mLS). Three behavioral tests in total were performed at 2-week intervals. In the first test, all animals were subcutaneously injected with 1.5 microg/kg estradiol benzoate (EB). Forty-four hours after EB, 0.5 mg progesterone (P) was injected and a behavioral test was started 4 h after P. These hormonal regimes were used in all tests. In the second test, 2 h before EB injection, 27-gauge cannulae filled with estradiol (E2) were inserted into the DRN, LS or CX through the guide cannulae and were kept there for 4 h. In the third test, cholesterol was implanted instead of E2 into these areas. In the first test, most females showed low levels of lordosis quotient (LQ) and most males showed no lordosis. In the second test, mean LQs in the LS or rLS groups of females increased but not in the DRN and CX groups. In the mLS group no increase of LQ was observed. When cholesterol was implanted in the third test, mean LQs in all groups were as low as in the first test. These results suggest the possibility that estrogen releases the inhibition when it acts on the LS, but not on the DRN female rats. On the other hand, inhibition in the male LS may not be released by the direct action of estrogen. PMID- 10364698 TI - Effects of experimental hypothyroidism on 5-HT1A, 5-HT2A receptors, 5-HT uptake sites and tryptophan hydroxylase activity in mature rat brain1. AB - The study was aimed at investigating the repercussions of deficiency in thyroid function with and without thyroid hormone (TH) replacement on the neurochemical entities which underly serotonin (5-HT) neutrotransmission, namely 5-HT1A, 5-HT2A receptors, 5-HT transporter and tryptophan hydroxylase (TPH) in the mature brain. Surgically thyroidectomized male Wistar rats received: (1) an iodine-free diet to produce severe hypothyroidism; (2) hormonal replacement with 15 microgram/kg/day of thyroxine (T4) for 21 days to normalize serum TH levels, or (3) hormonal replacement with 200 microgram/kg/day of T4 for 14 days to produce an excess of circulating THs. Sham-operated rats were used as controls. Neither hypothyroidism nor an excess in serum TH levels affected 3H-8-OH-DPAT binding to 5-HT1A receptors, 3H-citalopram binding to 5-HT transporter and TPH activity in various brain structures indicating that, in the mature brain, the presynaptic entities of 5-HT neurotransmission are resistant to large variations in TH levels. By contrast, hypothyroid rats had a significant decrease in Bmax of 3H-ketanserin binding to cortical 5-HT2A receptors compared to controls. Cortical 3H-ketanserin binding in thyroidectomized rats was normalized after replacement with low-dose T4. Excess serum TH levels in thyroidectomized rats did not produce any changes in cortical 5-HT2A receptors when compared to thyroidectomized rats with normalized TH levels. The present data suggest that the decrease in cortical 5 HT2A receptors is the main neurochemical event underlying the impairing effect of hypothyroidism on 5-HT neurotransmission. PMID- 10364699 TI - Thyroid hormones regulate the expression of somatostatin receptor subtypes in the rat pituitary. AB - Circulating TSH levels are increased in hypothyroidism and suppressed in hyperthyroidism. On the other hand, the hypothalamic hormone somatostatin suppresses basal and TRH-induced TSH release, an effect which is enhanced by thyroid hormones. To investigate whether the effects of thyroid hormones on TSH secretion may be mediated in part through alterations in the gene expression of pituitary somatostatin receptors (SSTR), 3-week-old male Sprague-Dawley rats were rendered hypothyroid with antithyroid drugs for 3 weeks. Total RNA extracted from anterior pituitaries were analysed for SSTR mRNA levels, using Northern blot hybridization. Compared to controls, hypothyroid rats had significantly lower pituitary mRNA levels of SSTR1 and SSTR2 (p < 0.0001 for both, n = 16); the reductions could be prevented by T4 supplementation (3 microgram/100 g body weight/day i.p.). In vitro studies using GH4C1 rat pituitary cells showed that the addition of T3 10(-8) M to cells cultured in charcoal-stripped bovine calf serum resulted in significant increases in mRNA levels of SSTR1 (p < 0.0001; n = 7) and the two transcripts of SSTR2 (p < 0.0005; n = 7). The increase for SSTR1 showed no further increase with higher doses of T3, but was time-dependent and could be seen consistently after 8 h of incubation. We conclude that thyroid hormones regulate the gene expression of SSTR subtypes in the pituitary, via a direct action on anterior pituitary cells. Changes in SSTR gene expression may contribute to the increase in circulating TSH levels in hypothyroidism. PMID- 10364700 TI - Cerebral salt-wasting syndrome: does it exist? AB - Cerebral salt-wasting syndrome (CSWS) has been regarded as a misnomer of the syndrome of inappropriate secretion of antidiuretic hormone (SIADH). We take the position that CSWS does exist and might be more common than SIADH. Differentiation between groups has been difficult because of overlapping signs, symptoms, and associated diseases. Euvolemia in SIADH and hypovolemia in CSWS may be the only contrasting variables. However, clinical assessment of extracellular volume is accurate in about 50% of these patients. Determination of serum urate and fractional excretion rates of urate can differentiate one group from the other. In both groups, hyponatremia coexists with hypouricemia and increased fractional excretion of urate. When the hyponatremia is corrected by water restriction, hypouricemia and elevated FEurate correct in SIADH but persist in CSWS. Persistent hypouricemia and elevated FEurate were commonly noted with pulmonary and/or intracranial diseases. The absence of intracranial diseases in some patients suggests that renal salt wasting might be a more appropriate term than CSWS. A review of renal/CSWS reveals three studies involving hyponatremic neurosurgical patients who had decreased blood volume, decreased central venous pressure, and inappropriately high urinary sodium concentrations in the majority of them, suggesting that CSWS was more common than SIADH in neurosurgical patients. Evidence for the presence of a plasma natriuretic factor in CSWS is presented. PMID- 10364701 TI - Cerebral salt-wasting syndrome. We need better proof of its existence. AB - It is widely believed that the cerebral salt-wasting syndrome (CSWS) exists as an entity distinct from the syndrome of inappropriate ADH secretion, and that it is characterized by evidence of severe renal salt wasting that results in volume depletion and hyponatremia. Proof of the existence of CSWS as an entity requires documentation of renal salt wasting and volume depletion. The present review has been undertaken to examine the evidence that the CSWS is a separate entity. In this effort, we have discussed various methods of documentation of volume depletion, and then reviewed reported cases of CSWS to determine whether volume depletion and renal salt wasting have been clearly demonstrated. Our review has led us to conclude that not one case of purported CSWS has demonstrated clear evidence of volume depletion and renal salt wasting. If renal salt wasting had been proven in these cases, we would conclude that the likely site of renal salt transport was the proximal tubule. The proximal site of salt transport defect has been suggested by the absence of hyperreninemia and hypokalemia, which would be a distinguishing feature of Bartter's syndrome and Gitelman's syndrome. PMID- 10364702 TI - Exercise coaching and rehabilitation counseling improve quality of life for predialysis and dialysis patients. AB - Advances in medical treatment have improved the rehabilitation potential of predialysis (P) and dialysis (D) patients, but deficits remain in their physical and vocational functioning. We studied 18 P (expected to begin dialysis in 6-12 months) and 18 D patients (on dialysis 1-5 years) for 1 year. Exercise coaching and rehabilitation counseling were provided at no cost for the first 6 months to half of each patient group (rehabilitation group = R); the other half were assigned randomly to controls (C). No R services were provided during 6 months of follow-up. PR walked further in 6 min at 6 months (+3.9 m) and 12 months (+4.1 m) than initially (p < 0.01). Hematocrit increased in R (p < 0.05), but not in C. Symptom scores were stable in D, worsened 21% in PC, and improved 15% in PR. Sickness impact profile scores were better in PR than PC at 6 months (p < 0.05) and 12 months (NS). Comorbidity correlated with symptoms (r = +0.34, p < 0.05), self-rated affect (r = -0.35, p < 0.05), and self-rated Karnofsky index of disability (r = -0.37, p < 0.05), but not with physician-rated affect or physician-rated Karnofsky index of disability. Thus, quality of life was stable or improved in PR, but declined in PC; PR benefited more than DR. Rehabilitation services are more beneficial before than after patients stabilize on dialysis, and quality of life monitoring should continue indefinitely. PMID- 10364703 TI - Potassium removal increases the QTc interval dispersion during hemodialysis. AB - This study was planned to clarify the mechanism(s) by which hemodialysis increases the QTc dispersion, a marker of risk of ventricular arrhythmias. To this aim, 10 uremic patients, without any relevant heart diseases, underwent two different types of hemodialysis schedules. In the first, 1 h of isolated high rate ultrafiltration preceded the standard diffusive procedure. In the second, during the first hour of standard bicarbonate hemodialysis, the decrease of plasma potassium concentration was prevented by increasing K+ concentration in the dialysate, according to its pre dialysis plasma levels. During the high rate ultrafiltration period, together with ECG signs of increased sympathetic nervous system activity and catecholamines secretion, the QTc dispersion did not change significantly. Instead, an evident increment was observed 1 h after the start of the diffusive hemodialysis, then slowly progressing until the end of the dialysis and finally returning to the pre dialysis values within 2 h after the end of the session. To the contrary, the increase of the QTc dispersion was totally blunted during a standard hemodialysis procedure in absence of plasma K+ decrease, but appeared again when the K+ dialysate fluid concentration was restored to 2 mmol/l. This study provides evidence that the increase of QTc dispersion occurring on hemodialysis is mainly related to the diffusive process, more precisely to the K+ removal. This is one more reason to focus attention on K+ removal rate especially when hemodialysis treatment is given in uremics affected by cardiac diseases with high risk of arrhythmias. PMID- 10364704 TI - Renal cell carcinoma and paraneoplastic IgA nephropathy. AB - Paraneoplastic nephropathy is rarely associated with human tumors. Little is known about the pathogenetic background of this relationship. To our knowledge, no conclusive study of the association of potentially 'immunogenic' renal cell carcinoma (RCC) and paraneoplastic nephropathy has been published. For this reason, we performed an immunohistochemical analysis of native resected kidneys of 60 patients with RCC, paying special attention to their pre- and postoperative records. Sixteen (27%) of the 60 tumor patients had immune complex nephropathy (11 IgA nephropathy [IgA NP] and 5 focal segmental glomerulosclerosis [FSGS]). Preoperative proteinuria and/or hematuria observed in 11 of 16 cases disappeared in 6 IgA NP patients within a 2- to 3-month follow-up after nephrectomy. Eleven of 16 tumors stained with the anti human immunoglobulin (IgA or IgM) of the same isotype as that present in glomerular immune complexes. In 3 IgA NP patients RCC associated von Hippel-Lindau (VHL) protein and IgA staining were found simultaneously in the tumor and glomeruli, with the clinical and laboratory findings disappearing after nephrectomy. Immune injury of the glomeruli due to a tumor-induced antigen-antibody response was demonstrated in these 3 IgA NP patients. PMID- 10364706 TI - Relationship between polymorphism in the angiotensinogen, angiotensin-converting enzyme or angiotensin II receptor and renal progression in Japanese NIDDM patients. AB - We determined the relationship between the gene polymorphism of angiotensinogen (AGT), angiotensin-converting enzyme (ACE), or angiotensin II receptor (AT1R) and the progression of diabetic nephropathy in a multicenter trial of ethnically homogeneous Japanese patients with non-insulin-dependent diabetes (NIDDM). Gene polymorphism of ACE I/D, AGT M235T and AT1R A1166C was determined by polymerase chain reaction amplification using allele-specific primers. Japanese NIDDM patients (n = 1,152) were selected from several diabetic clinics. All patients were divided into three groups as follows: (1) group I (n = 407): normoalbuminuric patients; (2) group II (n = 327): microalbuminuric patients, and (3) group III (n = 418): overt albuminuric patients. Clinical factors for investigation in all patients were the date of birth, gender, levels of urinary albumin excretion, findings of the ocular fundus, duration of diabetes, hemoglobin A1c and blood pressure. It appears that genetic polymorphisms in the renin-angiotensin systems, i.e. ACE or AT1R, may affect the progression to renal failure of patients (especially females) with NIDDM. PMID- 10364705 TI - Age, renal perfusion and function in island-dwelling indigenous Kuna Amerinds of Panama. AB - BACKGROUND/AIMS: Among possible contributors to a progressive fall in renal perfusion and function with increasing age, some hypotheses have invoked the rise in blood pressure that occurs with age, and a high-protein diet typical of urban cultures. Kuna Amerinds residing in isolated islands off the Panamanian Coast have a very low protein intake and show no tendency for blood pressure to rise with age, thus providing an opportunity to test these hypotheses. METHODS: We measured renal plasma flow and glomerular filtration rate (PAH and inulin clearance) in 16 Kuna Indians ranging in age from 18 to 86 years (51 +/- 6 years) who have resided on Ailigandi, an isolated Panamanian island for all of their lives. Inulin and PAH were infused with a battery-driven pump for 60 min, and a metabolic clearance rate used to calculate inulin and PAH clearance. For comparison, we employed identical techniques in 29 residents of Boston, ranging in age from 19 to 79 years (52 +/- 4 years), all normotensive and free of disease or medication use. Twenty-four were Caucasian. RESULTS: The Bostonian controls showed the anticipated fall in PAH clearance with age (y = 806 - 4.9 x; r = 0.82; f = 38.0; p < 0.0001). Our hypothesis was that the absence of a blood pressure rise with age and the low protein intake would flatten the slope relating renal perfusion to Kuna age. Our finding was a numerically steeper slope relating age and renal plasma flow in the Kuna (y = 936 - 6.48x; r = -0.81; p < 0.001). Filtration fraction rose with age in both populations, and again the rise was steeper in the Kuna. GFR in the Kuna, on the other hand, was very much higher at any age (139 +/- 4 ml/min/1.73 m2) than in Bostonians (112 +/- 3 ml/min/1.73 m2; p < 0.001). CONCLUSION: The findings are not in accord with the hypothesis that age-related changes in renal perfusion and glomerular filtration rate reflect an important contribution from blood pressure rise and a high protein intake, typical of modern, urban life. PMID- 10364707 TI - Correlation between cytomegalovirus infection and Raynaud's phenomenon in lupus nephritis. AB - Relationships between viruses and autoimmune diseases such as systemic lupus erythematosus (SLE) are still elusive. Recent reports demonstrated the association of some viral infections with peculiar clinical events in the general population, such as cytomegalovirus (CMV) with arterial damage and Parvovirus B19 (PV-B19) with hematologic abnormalities. We planned to look for this kind of viral imprinting in SLE, hypothesizing that traces of specific features of some viral infections might be found in some subsets of seropositive SLE patients. In 60 SLE patients recruited at our nephrologic center, serology for CMV, PV-B19, Epstein-Barr virus viral capsid antigen (EBV-VCA), Epstein-Barr nuclear antigen (EBNA) and Epstein-Barr virus early antigen (EBV-EA) was performed. chi2 and ANOVA were employed to compare the frequency and titers of antiviral antibodies in SLE patients with groups of transplant, hemodialysis and blood donor subjects. chi2, Fisher's test, Bonferroni and Scheffe's test were employed to compare the different biochemical/clinical features between seropositive and seronegative SLE patients. Univariate and multivariate analysis (logistic regression models) were employed to evaluate the odds ratio (OR) of different risk factors for vascular events (including Raynaud's phenomenon, deep venous thrombosis) and hematologic abnormalities (including severe anemia, leukopenia and thrombocytopenia). Anti CMV (82%), anti-PV-B19 (60%), anti-EBV-VCA (92%) and EBV-EA (45%) IgG antibodies were frequent in SLE, with higher prevalence in comparison with the blood donor group and higher titers in comparison with transplant and hemodialysis groups. CMV seropositivity was a highly significant risk factor for Raynaud's phenomenon (OR +alpha in univariate and multivariate analysis = 13.51 using a correction of 0.5 in case of a zero event), but not for venous vascular events (OR = 1.31). An increased though not significant risk factor was found for antiphospholipid antibodies (OR = 2.71, p = 0.19), while the presence of nephrotic syndrome during the follow-up was a significant protective factor (OR = 0.15, p = 0.035). There was no significantly increased OR for PV-B19 seropositivity in cases with severe anemia (OR = 2.09, p = 0. 29). No significant associations were found with the status of EBV reactivation. In conclusion, our results support the hypothesis that viral infection may imprint the course of SLE leading to specific clinical subsets (i.e. CMV and 'vascular' SLE, with more frequent Raynaud's phenomenon and a less frequent typical histological renal picture responsible for nephrotic syndrome). Further prospective studies are justified to validate these correlations, mainly dealing with associations between acute viral infections and vascular events, thus eventually leading to a better understanding of mutual relationships between viruses and SLE. PMID- 10364708 TI - Effect of vitamin E on adriamycin- induced nephrotoxicity at the ultrastructural level in guinea pigs. AB - It is known that Adriamycin, which is widely used in the treatment of various neoplastic conditions, exerts toxic effects in several organs. In this study, we have established that vitamin E has some beneficial effects on the kidney by protecting it from some of the toxicity induced by Adriamycin. A study was carried out which comprised one control group and two experimental groups of guinea pigs. In the experiment Adriamycin was administered either alone (group II) or together with vitamin E (group III). The results of groups II and III were compared with controls (group I). The kidneys were subsequently removed and examined by routine electron microscopic techniques. We found that vitamin E administered together with Adriamycin could reverse some of the degenerative changes caused by Adriamycin. PMID- 10364709 TI - High glucose induces a hypertrophic, senescent mesothelial cell phenotype after long in vivo exposure. AB - Previous studies, done using our mouse model for population analysis of the mesothelium, showed evidence indicating that in vivo, long-term exposure (up to 30 days) of the peritoneum to high-glucose (4.25% D-glucose) concentration dialysis solutions resulted in a hypertrophic mesothelial phenotype characterized by increased cell surface area, multinucleation, low proliferative capabilities, reduced cell viability, and enhanced enzymatic activity. These elements that define a senescent population of cells were not related to the pH of the fluid and its osmolality, or to the presence of buffer lactate. The present study was designed to explore the adverse effects of a lactate-free, filter-sterilized, high-D-glucose concentration solution (4.25%) at normal pH and prepared in Hanks' buffered salt solution after 2 h, 15 and 30 days of once a day intraperitoneal injection. Analysis of our observations indicate that in vivo exposure of the mesothelium to a high-glucose concentration induced a decreased density of the cell population, made up by larger and multinucleated cells, the viability of which was significantly lower than that observed in intact unexposed mice. The prevalence of mitosis showed an early and short-lived acceleration (up to 3 days), followed by values near zero during the rest of the follow-up period. So far, the main effect of the high-glucose concentration appears to result not from a mechanism of cytotoxicity, but from a substantial change in the life cycle of the exposed cell population, leading to their premature senescence and death in apoptosis. We hypothesize that this outcome may well be mediated by sustained oxidative stress derived from both a reduced production of scavengers, as well as the increased generation of oxygen-reactive species. PMID- 10364710 TI - Discordant evolution of asymptomatic proteinuria in identical twins. AB - We describe a pair of 17-year-old identical twin brothers with asymptomatic proteinuria, one of whom showed focal segmental glomerulosclerosis (FSGS) while the other showed immunoglobulin M (IgM) nephropathy. For each twin, audiological examination was normal. There was no family history of renal failure, deafness, or hematuria. HLA typing revealed an identical phenotype consisting of A25, A33, B44, B54, Cw1, Cw7, DR7 and DRB1. There is still controversy about whether minimal change disease, IgM nephropathy, and FSGS are discrete entities or different aspects of the same disease. The coexistence of IgM nephropathy and FSGS in identical twins suggests that the same genetic factors may be involved in the development of both diseases. However, although the brothers are identical twins, they had different eating habits and body weight. The twin who preferred to eat a protein-rich diet and who was heavier developed early proteinuria and manifested FSGS on renal biopsy. The discordant evolution of asymptomatic proteinuria in identical twins may provide a clue for the existence of environmental factors on the progression from IgM nephropathy to FSGS. Therefore, this report provides indirect support for the hypothesis that IgM nephropathy and FSGS represent different aspects in the spectrum of a single disease. PMID- 10364711 TI - Yellow nails and minimal change nephrotic syndrome. AB - We report a case of a 38-year-old man showing the yellow nail syndrome in association with minimal change nephrotic syndrome. Treatment with prednisone and vitamin E resulted in complete resolution of the nephrotic syndrome and slow improvement of the yellow nails, respectively. Although the rare yellow nail syndrome has been described in association with renal disease, this is the first report of the association of this syndrome with minimal change nephrotic syndrome. PMID- 10364712 TI - Positive Coombs test in pneumococcus-associated hemolytic uremic syndrome. A review of the literature. PMID- 10364713 TI - Diastolic dysfunction increases the frequency of ventricular arrhythmia in hemodialysis patients. PMID- 10364714 TI - Cyclosporin A nephrotoxicity in rats with genetically fixed hypertriglyceridemia. PMID- 10364715 TI - Elevation of serum IgA is of little diagnostic utility in patients with type 2 diabetes mellitus and nephropathy. PMID- 10364717 TI - What are surrogate outcome measures and why do they fail in clinical research? AB - Surrogate outcome measures have been of interest in clinical research for a long time because of their potential ability to save time and cost. The literature, however, has numerous examples of failed surrogate measures. The definition of a surrogate outcome measure, its consideration in clinical stroke research and some of the potential mechanisms for failure of surrogate measures will be discussed. PMID- 10364716 TI - Close association between HCV infection and cryoglobulinemia in patients on chronic hemodialysis. PMID- 10364718 TI - Validity of telephone interview data for vascular disease risk factors in a racially mixed urban community: the Northern Manhattan Stroke Study. AB - The aims of our study were to assess the validity and reliability of a telephone survey instrument designed to measure vascular disease risk factors and to assess whether these measurements were influenced by age, gender, race/ethnicity, or other sociodemographic variables. Subjects were sampled and interviewed using random digit dialing methodology from the multiethnic community of northern Manhattan. For the validity study, 261 consecutive subjects were clinically assessed in-person within 60 days of the telephone interview. A retest reliability study of the telephone interview was conducted in 92 randomly selected subjects within 30 days of the initial interview. The telephone interview instrument had a sensitivity of more than 55% and a specificity of 74% or greater for various vascular disease risk factors. Sensitivity, specificity, and positive predictive value did not vary significantly or systematically among whites, blacks, and Hispanics, but subjects with access to health care were more likely to provide valid data. The reliability substudy indicated a good reliability for the telephone interview. These results support the validity of telephone interviews for estimating the prevalence of vascular risk factors in urban populations. PMID- 10364719 TI - Low cholesterol as a risk factor for primary intracerebral hemorrhage: A case control study. AB - We performed a case-control study to assess the relationship between primary intracerebral hemorrhage (ICH) and low serum cholesterol. Prospectively recruited, fully evaluated patients with ICH were compared to two independent control groups, one based in a primary care practice and one population-based. Low cholesterol was defined by the sex-specific lowest quintile of the primary care controls. The proportion of ICH cases with low cholesterol >3 months posthemorrhage was significantly greater than in controls (42 vs. 20% in either control group, p < 0.01). Subgroup analysis showed an overrepresentation of low cholesterols in probable hypertensive hemorrhage (47%, p < 0.05) but not in probable cerebral amyloid angiopathy (27%, p = 0.5). Low cholesterol increased the odds for hemorrhage 2.25-fold (1.12-4.50) after adjustment for age and apolipoprotein E genotype. These data confirm an increased risk for primary ICH associated with low cholesterol, a relationship that may apply specifically to hemorrhages from hypertensive vasculopathy. PMID- 10364720 TI - Association of cigarette smoking with amyotrophic lateral sclerosis. AB - We explored the relationship between amyotrophic lateral sclerosis (ALS) and cigarette smoking in a case-control study conducted in New England from 1993 to 1996. Recently diagnosed ALS cases (n = 109) were recruited from two major referral centers. Population controls (n = 256) were identified by random telephone screening. Data were analyzed by logistic regression. After adjusting for age, sex, region and education, ever having smoked cigarettes was associated with an increase in risk for ALS (odds ratio 1.7; 95% confidence interval 1.0 2.8). Average cigarettes smoked per day, years smoked and pack-years were all greater in cases than controls, but dose-response trends were not observed. Similar numbers of cases and controls had ever used alcohol, and only a small, nonsignificant association of drinks per month with ALS was observed. The association of cigarette smoking with ALS was not affected by adjusting for alcohol use. In contrast, the weak relationship of ALS with alcohol use was apparently due to confounding by smoking. PMID- 10364721 TI - Study of HLA as a predisposing factor and its possible influence on the outcome of multiple sclerosis in the sanitary district of Calatayud, northern Spain. AB - The relationship between multiple sclerosis (MS) and the HLA antigens DR2 and DQ1 is well recognised, but, in Spain, it has not been clearly defined. The aim of our study was to investigate the relationship between MS and HLA antigens in the sanitary district of Calatayud, northern Spain, and to correlate these antigens with the progression of the disease. Thirty-four patients were selected from a long-term (October 1990 to July 1996) prospective survey in the region where there was a prevalence rate of 58 per 100,000 population. The HLA antigens were determined in 31 patients. A control group of 895 people of Caucasian race was recruited from the same population. We performed serologic tests on all participants. Nucleotide typing was carried out in DR2-positive patients. The most frequent antigens in excess in MS were: A19 (odds ratio, OR: 2.29, p = 0.04), B5 (OR: 2.85, p = 0.02), B41 (OR: 7.65, p = 0.04), CW7 (OR: 3.4, p = 0.004), DR6 (OR: 6.18, p = 0.0001) and DR10 (OR: 3.4, p = 0. 004). The DR2 antigen was also more frequent in MS patients (39%) than in controls (19%; OR: 2.69, p = 0.01). All positive DR2 patients showed the DR15(2) split but not the DR16(2) split. The frequency of antigens CW4 and DR1 was lower in MS patients than in controls. The CW4 antigen was detected in 12% of the patients and in 33% of the controls (OR: 0.28, p = 0.04). The DR1 antigen was found in 20% of the controls and in none of the MS patients (OR: undefined, p = 0.01). The DQ1 antigen was observed in 68% of the patients and in 50% of the controls (OR: 2.1, p = 0.07). We did not find any relationship between HLA antigens and progression of the disease. Although we found that DR2 antigen is linked to MS, we also found other antigens related to the disease. This suggests a genetic heterogeneity in our geographic area. We also concluded that the DR1 antigen may play a protective role, as it was detected in 20% of the controls and in none of the MS cases. PMID- 10364722 TI - Stroke survival after discharge from an acute-care hospital. AB - BACKGROUND AND PURPOSE: Survival after a stroke is likely to be best for patients well enough to be sent home but the relative risk of dying if patients do not qualify for a home discharge has not been well studied. We investigated the survival prognosis after an initial stroke depending on the facility to which the patient was discharged after an acute initial stroke. METHODS: All patients were enrolled between July 1, 1987, and August 1, 1989, and were followed up to 4 years (mean of 24 months) until death, second stroke, or the end of the study. RESULTS: Among 662 patients who were discharged alive after hospitalization with an initial stroke, 128 (19%) went to a nursing home, 17(3%) to a short-term hospital, 140 (21%) to a rehabilitation facility, and 375 (57%) went home (discharge destination unknown for 2 patients). Compared to patients sent home after taking age, sex, selected baseline comorbidities, length of hospital stay, and neurological deficits into consideration, results from Cox proportional hazards model indicated that patients sent to a nursing home had 2.6 times greater risk of dying (95% CI = 1.81-4. 15) while those who were discharged to a rehabilitation facility had a death hazards ratio of 1.1. CONCLUSIONS: Mortality was greatest in the early months after discharge and decreased thereafter. Since the analysis was adjusted for age, sex, comorbidity, length of hospital stay, and number of neurological deficits, quality of care in a nursing home setting may account for the mortality difference but other factors such as social support network and living will instructions also need to be investigated. PMID- 10364723 TI - Prevalence of multiple sclerosis in Gypsies and Bulgarians. AB - There are occasional reports about the low prevalence of multiple sclerosis among Gypsies. To verify these reports of low prevalence of multiple sclerosis among Gypsies compared to the white population in Bulgaria, an epidemiological study was conducted in two small regions of Bulgaria. All patients with clinically or laboratory-supported, definite multiple sclerosis according to Poser's criteria were personally interviewed and examined. The study was begun on January 1, 1997 and March 31, 1998 was selected as prevalence day. For the white population, the prevalence ratio of multiple sclerosis per 100,000 population was 44.9 in the first region and 44.4 in the second. The prevalence of multiple sclerosis in Gypsies was found to be 19.1/100,000 in the first region and 18.4 in the second. It is concluded that multiple sclerosis is less common in Gypsies than in whites living in the same areas. PMID- 10364724 TI - Lycanthropy in depression: two case reports. AB - Two cases of lycanthropy presenting as part of a depressive disorder are described. The patients responded favorably to pharmacotherapy. In both cases, a positive history of dog bite influenced the presentation of symptoms. The authors speculate whether the defense of identification with the aggressor was operative. PMID- 10364725 TI - Lycanthropy: new evidence of its origin. AB - Two cases of lycanthropy will be described. Its possible aetiology and psychopathology will be discussed. In the first case there is clear evidence of an organic origin of the syndrome which is reported for the first time. PMID- 10364726 TI - Psychiatric morbidity in disintegrative psychosis and infantile autism: A long term follow-up study. AB - In order to study the validity of disintegrative psychosis (DP), the authors compared 13 patients given this diagnosis in childhood with a control group of 39 patients with infantile autism (IA) matched for sex, age, IQ and social class on measures of psychiatric morbidity. Almost the same proportion of the two groups had been admitted to a psychiatric hospital during a 22-year follow-up period. However, there was a slight tendency (statistically nonsignificant) for the DP group to utilize the psychiatric health care system more frequently than the IA group. They had more admissions and stayed longer in hospital than patients with IA suggesting that they had more psychiatric symptoms than the IA group. The original IA diagnoses were confirmed fairly consistently during the follow-up period, while the DP group was given more heterogenous diagnoses. No diagnosis of schizophrenia was made in either group. PMID- 10364727 TI - Terminator: An unusual form of self-mutilation. PMID- 10364728 TI - Clinical homogeneity of DSM-IV schizophrenic disorders. AB - We investigated the lifetime fulfillment of five subcriteria of diagnostic criterion A in 94 inpatients with a definite diagnosis of DSM-IV schizophrenic disorder. Among the five diagnostic features captured by criterion A, only delusions were found to be almost universal, whereas of the remaining four, only hallucinations and negative symptoms occurred in more than half of all the cases. Overall, almost 60% of the 28 possible patterns of combination among the five subcriteria were found to be fulfilled although 60% of the cases were accounted for by only 4 patterns, which were also identified by means of a cluster analysis, as accounting for the totality of the cases. The first cluster was characterized by the fulfillment of subcriteria 1,2 and 5, the second by subcriteria 1 and 2, the third by subcriteria 1 and 5 and the fourth by subcriteria 1,2, and 3. The substantial clinical heterogeneity of the DSM-IV category of schizophrenic disorders with respect to their diagnostically characteristic features captured in criterion A is traced to the polythetic character of its five subcriteria. PMID- 10364729 TI - Long-term course and outcome of severe postpartum psychiatric disorders. AB - Thirty-nine women who had suffered from a severe first-episode postpartum psychiatric illness were re-examined after a period of 6-26 years (averaging 12.5 years). Diagnoses were established according to ICD-10 and Leonhard's classification, revealing a marked predominance of cycloid psychoses (54%) according to Leonhard. There was no evidence of the nosological independence of postpartum psychosis. Only 4 patients (10%) had never recovered fully since the onset of the illness. In contrast, 6 patients had undergone a monophasic course without any further psychopathology. In 20 cases (51%) the illness had run a multiphasic course. The average number of episodes per patient was 2.5 (range 2 6). The course was not determinable in 4 patients (10%). Nineteen women (49%) had 22 further deliveries after the first manifestation of the illness. The frequency of a relapse in connection with further pregnancy or delivery was 50%. Applying the Strauss-Carpenter Outcome Scale, we found a favourable outcome for the total sample with a mean value of 14.1 (SD = 2.6). The vast majority of patients (75%) showed no persistent alterations. Our findings provide further evidence of a favourable prognosis of severe postpartum psychiatric disorder despite the remarkably high rate of puerperal relapses. PMID- 10364730 TI - Capgras syndrome and functional psychosis in 2 survivors of torture. AB - Functional psychosis has only rarely been described in context with extreme stressors, most studies focusing singularly on posttraumatic stress disorder symptoms. We report for the first time the case histories of 2 patients suffering from Capgras syndrome along with schizoaffective disorder and posttraumatic stress disorder after prior experience of prolonged torture. Interaction of personal life experience and psychiatric disorder are proposed as factors resulting in persistent changes in perception and affect. PMID- 10364731 TI - Anhedonia in the deficit syndrome of schizophrenia. AB - Previous studies have shown that anhedonia characterizes the deficit syndrome of schizophrenia. Anhedonia is also one of the main symptoms of the depressive state. The purpose of this study was to examine the relationships between anhedonia and depression in the deficit syndrome of schizophrenia. Self evaluations of anhedonia and depression were performed by three groups of subjects (32 deficit schizophrenics, 32 major depressives, 35 healthy subjects) matched for sociodemographic variables. Deficit schizophrenics and major depressives are more anhedonic than controls, but there is no difference between the two study groups. Contrarily to what is evidenced for major depressives and for healthy subjects, the depressive symptomatology correlates with anhedonia in deficit schizophrenics. When deficit schizophrenics are dichotomized into depressed versus non-depressed patients, no difference is observed concerning anhedonia. These results suggest that anhedonia in the deficit syndrome of schizophrenia has no specificity but appears independent of coexisting depression and covaries with several characteristics of depression (retardation, cognitive distortions). Our results support the hypothesis that the deficit syndrome of schizophrenia could constitute a non-depressive mood disorder. PMID- 10364732 TI - An unusual case of delusional misidentification: 'delusional hermaphroditism'. AB - A 44-year-old man presented with an unusually colourful and complex form of delusional misidentification. He demonstrated Fregoli syndrome, environmental reduplication and the delusional belief that he and a female friend had been incorporated into the same body. This latter phenomenon has not been described before and adds a new dimension to the rich variety of these syndromes. It is suggested that the term 'delusional hermaphroditism' be used to describe this form of delusional misidentification. PMID- 10364733 TI - The lung in the immunocompromised patient. Infectious complications part 2. AB - Pulmonary infections decisively contribute to morbidity and mortality in immunocompromised patients. Bacterial, mycobacterial and infections with Pneumocystis carinii have been reviewed in an article in the last issue of Respiration. In this review, viral and fungal pulmonary infections are discussed in HIV-positive patients and in patients treated with high-dose chemotherapy, stem cell or solid-organ transplantation. PMID- 10364734 TI - A role for natriuretic peptides as well as endothelins in the hypoxic lungs of patients with chronic obstructive pulmonary disease. PMID- 10364735 TI - Reproterol: beta-2-agonist, theophylline, or both? PMID- 10364736 TI - Plasma endothelin-1 level in chronic obstructive pulmonary disease: relationship with natriuretic peptide. AB - BACKGROUND AND OBJECTIVE: Endothelin-1 (ET-1) is a potent vasoconstrictor peptide produced by the vascular endothelium. The purpose of this study was to elucidate the pathophysiological role of ET-1 in patients with pulmonary hypertension secondary to chronic obstructive pulmonary disease (COPD). METHOD: We measured plasma ET-1 levels during right heart catheterization both at rest and during exercise on room air and while breathing oxygen in patients with COPD. In addition, we simultaneously measured plasma levels of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP). RESULTS: Plasma ET-1 levels at rest were significantly higher in 21 patients with COPD than in 16 control subjects (p < 0.001). For COPD patients, there was no correlation between the plasma ET-1 level and pulmonary arterial pressure or pulmonary vascular resistance at rest. On the other hand, there was a significant negative correlation between plasma ET 1 level and mixed venous oxygen tension (r = -0. 503, p < 0.05). Also, the plasma ET-1 level was positively correlated with those of ANP (r = 0.540, p < 0.05) and BNP (r = 0. 533, p < 0.05) at baseline. Oxygen administration significantly decreased plasma ET-1 levels at rest (p < 0.05). Plasma ET-1 levels did not change significantly with exercise despite the progression of pulmonary hypertension and hypoxemia. In contrast, plasma ANP and BNP levels both increased markedly with exercise (p < 0.01). CONCLUSION: We conclude that in patients with COPD, the plasma ET-1 level is not affected by acute progression of pulmonary hypertension and hypoxemia during exercise, and persistent hypoxemia may be associated with an increase in the plasma ET-1 level. In addition, our findings suggest that ANP and BNP may modulate the pulmonary vascular tone by interacting with ET-1 in these patients. PMID- 10364737 TI - Reproterol--A monomolecular combination of orciprenaline and theophylline: novel aspects of its mode of action in asthma. AB - BACKGROUND AND OBJECTIVE: Reproterol is a monomolecular combination of orciprenaline and theophylline used as beta-adrenergic agonist to induce bronchodilation in bronchial asthma. Since the mechanism of action of reproterol has not been investigated so far, its potential anti-inflammatory activity in asthma remains still unknown. Therefore, we have studied in vitro whether the theophylline component of the reproterol molecule might enhance the stimulatory effect of the beta-adrenoceptor on cAMP production resulting in suppression of inflammatory mediator production. METHODS: The effects of reproterol, orciprenaline and theophylline (10(-9)-10(-5) M) on spontaneous cAMP (5 x 10(4) cells/30 min)- and on LPS (10 microg/ml)-stimulated LTB4 production (10(5) cells/4 h) were determined in normal monocytes in vitro. RESULTS: Production of cAMP (n = 9) was significantly augmented in a dose-dependent manner by orciprenaline (30 +/- 8%) and theophylline (28 +/- 10%), but mostly by reproterol (127 +/- 8%) at 10(-5) M. Despite incubation with propranolol, significant stimulation of cAMP production was notable following reproterol therapy. Production of LTB4 was significantly inhibited by reproterol (-48 +/- 14%) and less by theophylline (-28 +/- 10%), but was stimulated by orciprenaline (+20 +/- 8%) at 10(-5) M. CONCLUSION: We conclude that reproterol exerts a strong stimulatory effect on monocyte cAMP production and a suppressive effect on LTB4 production possibly due to a synergistic mode of action on adenylate cyclase activity and inhibition of phosphodiesterases. More clinical studies in bronchial asthma will be needed to determine whether these results may translate into clinically relevant effects. PMID- 10364738 TI - Repetitive hemodilution in chronic obstructive pulmonary disease and pulmonary hypertension: effects on pulmonary hemodynamics, gas exchange, and exercise capacity. AB - BACKGROUND: In cor pulmonale associated with severe chronic obstructive pulmonary disease (COPD), disturbances of pulmonary microcirculation may contribute significantly to hypoxemia, pulmonary hypertension, and exercise intolerance. OBJECTIVE: It was tested whether reduction of blood viscosity induced by repetitive hemodilution might improve pulmonary hemodynamics and oxygen uptake. METHODS: Seven patients with stable COPD (forced expiratory volume in 1 s 33 +/- 3 % of predicted, means +/- SE) and pulmonary hypertension were phlebotomized 5-6 times over a period of 3 months with substitution of 6% hydroxyethyl starch (molecular weight 40, 000). This resulted in a stepwise reduction of the hematocrit from 53.3 +/- 2.6 to 45.8 +/- 3.1% and a reduction of whole blood viscosity from 9.8 +/- 0.6 to 8.8 +/- 0.7 mPa x s at a shear rate of 2.0 s-1. Before and after the treatment period, patients underwent cardiopulmonary exercise testing and right heart catheterization. RESULTS: Mean pulmonary artery pressure (PAm) decreased from 30 +/- 3 to 22 +/- 2 mm Hg and arterial oxygen partial pressure (PaO2) increased from 63.2 +/- 2.2 to 71.8 +/- 3.7 mm Hg at rest. During peak exercise, PAm decreased from 59 +/- 7 to 53 +/- 7 mm Hg and PaO2 increased from 54.0 +/- 5.7 to 63.2 +/- 2.4 mm Hg after hemodilution. Peak oxygen consumption rose from 573 +/- 84 to 750 +/- 59 ml x min-1, corresponding to an increase in cardiac index from 4.25 +/- 0.5 to 5.88 +/- 0.76 liters x min-1 x m-2. Pulmonary vascular resistance fell from 345 +/- 53 to 194 +/- 32 dyn x s x cm-5. The patients' peak exercise capacity increased from 9.2 +/- 2. 0 before to 13.5 +/- 3.2 kJ at the end of the study (p < 0.05 for all differences, paired t test). CONCLUSION: The findings suggest that a prolonged improvement of pulmonary microcirculation by reducing blood viscosity may improve pulmonary gas exchange, central hemodynamics, and exercise tolerance in patients with severe COPD and pulmonary hypertension. PMID- 10364739 TI - Antineutrophil cytoplasmic antibodies in diffuse panbronchiolitis. AB - BACKGROUND: There are some reports of the coexistence of chronic suppurative lung diseases such as cystic fibrosis and systemic vasculitis. Diffuse panbronchiolitis has the same characteristics as chronic recurrent sinopulmonary infection and respiratory bronchiolitis. METHODS: Serum samples from 30 patients with diffuse panbronchiolitis and 57 patients with other pulmonary diseases were tested to find the titer of myeloperoxidase antineutrophil cytoplasmic antibodies (MPO-ANCA). RESULTS: We found MPO-ANCA positivity in 4 patients with diffuse panbronchiolitis but not in those with other pulmonary diseases. CONCLUSIONS: Our findings show that MPO-ANCA is positive in some patients with diffuse panbronchiolitis. Careful attention should be paid to the combination of chronic pulmonary infection and various vasculitis. PMID- 10364741 TI - Static lung volumes: reference values from a Latin population of Spanish descent. AB - BACKGROUND AND OBJECTIVES: The aim of this study was to develop a set of prediction equations and 90% confidence intervals for static lung volumes using the multibreath helium equilibration method from a sample of asymptomatic Caucasian subjects of Spanish descent. Moreover, these equations were compared with those of previous studies. METHODS: Measurements of static lung volumes using techniques recommended by the American Thoracic Society and the European Community for Steel and Coal were carried out on a selected sample of 591 healthy nonsmoking volunteers (305 men and 286 women) aged 18-88 years, living in the metropolitan area of Valencia, on the east coast of Spain. Multiple regression analysis using height, age and weight as independent variables were used to provide predicted values for both sexes. These reference values were compared with other sets of prediction equations reported in the literature using an independent sample of 69 subjects (32 men and 37 women). RESULTS: Simple linear regression equations using age, height and body weight predicted all the subdivisions of lung volumes (vital capacity, expiratory reserve volume (ERV), inspiratory capacity, functional residual capacity (FRC), residual volume (RV), total lung capacity (TLC), FRC/TLC and RV/TLC) as well as more complex equational models. The distribution of residuals fulfilled the assumptions of multiple regression analysis (independence, homoscedasticity and Gaussian distribution of residuals), except for ERV, using simple linear models. The derived equations did not differ significantly from most of the previously reported equations and were usually superior in their ability to predict the lung volumes. CONCLUSIONS: The use of the present prediction equations is recommended in the Latin population of Spanish descent and in populations with similar Caucasian characteristics. PMID- 10364740 TI - The number of interleukin 1 receptors on lung fibroblasts in patients with idiopathic pulmonary fibrosis. AB - BACKGROUND: We investigated the expression of the IL-1 receptor on lung fibroblasts in patients with idiopathic pulmonary fibrosis (IPF) the histology of which was diagnosed as usual interstitial pneumonia. METHODS: 125I-labeled IL 1alpha bound to lung fibroblasts in a specific and saturable manner in lung fibroblasts obtained from both IPF patients and control subjects. RESULTS: Scatchard plot analysis revealed a single type of binding site in both groups. In total, 8,600 +/- 1,450 (mean +/-SD) sites/cell with a dissociation constant (Kd) of 1.83 +/- 0.28 (mean +/-SD) nM in patients with IPF and 7,920 +/- 1,470 sites/cell with a Kd of 2.43 +/- 0.49 nM in control subjects were counted. Furthermore, affinity cross-linking experiments revealed that the IL-1 receptor on lung fibroblasts in patients with IPF and control subjects had a molecular size of 80 kD. CONCLUSION: In conclusion, type I receptor was present on human lung fibroblasts, but there were no differences in number or affinity between IPF patients and controls. PMID- 10364742 TI - Maximum static respiratory pressures in healthy elderly men and women: issues of reproducibility and interpretation. AB - BACKGROUND: Respiratory muscle strength is assessed using the static pressure generated at the mouth during a maximal inspiratory or expiratory effort [PImax and PEmax, respectively (MSRPs)]. Interpretation of MSRPs relies upon comparison with 'normal' values, but MSRPs show very weak associations with predictors such as physical characteristics. The influence of habitual physical activity upon MSRPs remains undefined. OBJECTIVES: We examined measurement reproducibility, as well as the influence of physical characteristics and habitual physical activity upon MSRPs in healthy elderly people. METHODS: MSRPs were assessed in 41 healthy subjects using a portable mouth pressure meter on two occasions, 1 week apart. Physical activity was assessed in 10 subjects by diary record. Pearson product moment correlation coefficients were used to assess the association of MSRPs with other measured variables. RESULTS: There was good measurement reproducibility of MSRPs, with coefficients of reproducibility of 10.2 and 12.8% for PImax and PEmax, respectively. MSRPs showed statistically significant negative correlations with age, but correlations with physical characteristics were poor. In contrast, MSRPs were highly correlated with physical activity. CONCLUSIONS: We conclude that MSRPs can be measured reproducibly and that they decline with advancing age. Physical characteristics are not good predictors of MSRPs; this may be due to a strong confounding influence of physical activity making interpretation of measurements problematic. We suggest that the poor predictive power of physical characteristics indicate that reference to 'normal' values be made with caution and that it may be more appropriate to consider functional interpretations of MSRPs based upon factors such as lung and chest wall elastance. PMID- 10364743 TI - Decrease in lung volume depends on end-expiratory pressure in a rabbit model of airway lavage. AB - BACKGROUND: In the past, the rabbit model of repeated airway lavage has been extensively used to induce a decrease in lung volume accompanied by impairment in lung mechanics and gas exchange. OBJECTIVES: The rationale of our study was to investigate the influence of different end-expiratory pressure (EEP) levels (0.4 4.2 cm H2O) on changes in functional residual capacity (FRC) and the efficacy of lavages administered. METHODS: Forty-five rabbits were subjected to 2-8 lavages with 20 ml/kg warm normal saline until arterial/alveolar oxygen tension (a/A ratio) had decreased to 3 mg/dL on admission in ICU, or hyperbilirrubinemia > 2.5 mg/dL. Parenteral nutrition (TPN), consisting of 1.4 g AA + Lipids 1.3 g + carbohydrates 4 g/kg/d, (either glucose or FGX at random) was administered. Basal levels and days 1st, 4th and 10th plasma glucose, triglycerides, cholesterol, uric acid were determined, and blood venous gases as well. Capillary glycemia was measured every 6 hours and insulin given if glucose levels rose above 180 mg/dL. STATISTICS: Fisher's exact test; Student t-test; Mann-Whitney test. Data as mean and SD. RESULTS AND CONCLUSIONS: During 48 months, 119 patients admitted in the ICU (72 with pneumonia and 47 with pancreatitis) were included. In pneumonia, tolerance was similar with both intakes; glycemia was kept at the same level in both, but the amount of insulin given was significantly more in those patients fed on glucose (p < 0.05). Nevertheless, resting blood glucose and triglyceride levels were higher in pancreatitic patients, and more insulin was required. Those on FGX had lower triglyceride plasma levels (p < 0.05) and less insulin was given throughout the study. Glycemia was kept lower though no statistical significance was reached (p < 0.1). No hyperuricemia nor lactic acidosis was found. PMID- 10364785 TI - Small bowel adaptation with growth hormone and glutamine after massive resection of rat's small bowel. AB - The use of glutamine (GLN) and growth hormone (GH) improves intestinal adaptation in short bowel syndrome (SBS). The present study aimed to assess the effect of a diet rich in glutamine and the use of GH on intestinal adaptation in experimental SBS. 80 Wistar rats (240 g) were randomized into 6 groups: 1) RGLN (20)--95% small bowel resection and fed on GLN diet; 2) RGLNGH (20)--95% SBR, GLN diet and GH; 3) RC (10)--95% SBR and fed on a low GLN control diet (C); 4) RCGH (10)--95% SBR and C diet and GH; 5) TAGLIN (10)--intestinal transection and anastomosis (Ta) and fed on a GLN diet; 6) TAGLNGH (10)--Ta and GLN diet and GH. GH was given SC at a dose of 0.14 mg/kg/day. The rats were weighed daily and nitrogen balance was made. Rats were sacrificed after 15 days and mucosa cell proliferation was studied with PC10 antibody. Statistical analysis was performed. All SBR rats lost weight as compared to their initial weight (8% to 13%). GH improved Ta rats weight (18.98 x 5.04%). The use of GLN diet and GH improved nitrogen balance and bowel growth on SBR groups, as compared to controls, but not cell proliferation. In conclusion, the use of GLN enriched diet and GH improves intestinal adaptation after massive resection of the small bowel in rats. PMID- 10364786 TI - [Bioelectric impedance in the nutritional evaluation of mentally deficient quadriplegic adults]. AB - Severely mentally retarded persons with cerebral palsy show a high rate of acute and chronic malnutrition. Without discarging other factors which might be at play, caloric intake deficiency stemming from the difficulties involved in being fed appear to play a crucial role in its etiology. In the assessment of these disorders, anthropometry is limited by the lack of adequate reference values and by the difficulty or impossibility of measuring height using the conventional method. The purpose of this study is to see how changes in body composition brought about by an increase in dietary caloric intake are perceived by both anthropometry and biolectrical impedance analysis. To do so, 13 subjects were selected from a group of 203 severely mentally retarded persons made up of 25% cerebral palsy patients and 13% quadriplegics. The 13 subjects were fed orally without tubes and all had tricipital skinfolds of less than P25. All 13 were given a 25% caloric increase over the regular diets for a period of two months. This increase was provided by means of a polymeric, normoproteic, hypercaloric preparation. Weight and brachial perimeter showed significant increases coinciding with the administration of the supplement. Theses increases were not noticeable two months after discontinuing the supplement. There were no significant changes in tricipital and subescapular skinfolds, muscular circumference of de mid-arm, resistance, reactance, or the total body an extracellular water calculated from the latter two values. From these results we deduce that biolectrical impedance analisys offers no advantages over anthropometry in monitoring the nutritional status of quadriplegics. PMID- 10364787 TI - [Parenteral nutrition in a case of renal insufficiency due to amyloidosis]. PMID- 10364788 TI - [Neologisms, anglicisms and barbarisms in "AIDS-speak"]. PMID- 10364789 TI - [Practical considerations respecting the clinical use of serum cystatin C as a marker of glomerular filtration]. AB - A curvilinear relationship of potential type was observed between serum concentrations of cystatine C and creatinine in 187 patients attended at our Nephrology Department. This relationship became linear when considering the reverse values, but the estimation error was much higher than what is clinically acceptable, which prevents the interconversion of values. In 78 critical patients, in whom an antibiotic therapeutic monitoring was being carried out, serum creatinine concentrations in the reference range were observed to be associated with pathologic concentrations of cystatine C. In 22 regularly hemodialysed patients, using Hemophan or Cuprophan membranes, increments of approximately 5% for cystatine C and beta 2-microglobuline in post-dialysis samples were observed. The chronic administration of inducer antiepileptic drugs, capable of modifying the concentration of different plasmatic proteins and reducing the serum concentration of creatinine, does not seem to limit the applicability of cystatine C as marker for glomerular filtration. The routine measurement of cystatine C seems to be warranted for estimating the glomerular filtration rate in patients in whom creatinine synthesis might be impaired. PMID- 10364790 TI - [Transient blood surface antigens of hepatitis B in patients on hemodialysis]. AB - BACKGROUND: The presence of surface antigen of hepatitis B (HBsAg) virus in serum from recently vaccinated adults has been scarcely investigated. In this work, after the detection by chance of seven HBsAg-positive patients on hemodialysis who reported recent hepatitis B vaccination, a prospective study was undertaken to verify the presence and duration of post-vacunal antigenemia. PATIENTS AND METHODS: Nineteen non-selected patients on hemodialysis were followed for serologic markers of hepatitis B, after receiving a dose of the recombinant vaccine (Engerix B) according to their vaccination schedule. Enzyme-immunoassay techniques were used for the study of serologic markers, and the reactivity of HBsAg was confirmed by means of a neutralization assay with specific anti-HBs antibodies. RESULTS: After the administration of one vaccine dose, 31.5% of patients were HBsAg positive at least once. Antigenemia was identified more frequently 2 to 4 days (83.3%) after immunization. In all cases antigenemia was transient and had cleared after 11 days of vaccination. The follow-up of serologic markers revealed the absence of infection with virus B. Only 16.6% of patients with transient antigenemia responded to vaccination (titer of anti-HBs > or = 10 mIU/ml), while the corresponding percentage in the group of HBsAg negative patients was 69.2% (p < 0.05). All patients were HCV and HIV negative. CONCLUSIONS: A high frequency of post-vacunal antigenemia is reported in patients on hemodialysis, in absence of virus B infection as well as the possible relationship between the presence of transient antigenemia and the non-responder status. PMID- 10364791 TI - [The association of hematocrit with blood pressure and left ventricular mass in subjects over 55]. AB - OBJECTIVE: To establish the association between hematocrit, blood pressure (BP), and left ventricular mass (LVM) in persons older than 55 years. PATIENTS AND METHODS: One hundred patients older than 55 years (46 males and 54 females) with a wide range in BP were studied on an ambulatory basis. The following parameters were determined: hematocrit, casual BP (CBP), 24-h ambulatory blood pressure monitoring (ABPM), left ventricular mass index (LVMI) determined by echocardiography (according to the Devereux's formula), cardiac output (CO) (determined by cardiac echocardiography), peripheral vascular resistances (PVR) (determined according to mean blood pressure and cardiac output), and levels of creatinine in plasma and sodium in 24-hour urine. RESULTS: Mean age of patients was 65 years (range 55 to 83 years). Twenty-two percent of patients were smokers (17 males and 5 females) with a mean of 13 +/- 6 cigarettes/day. Mean hematocrit value was 41.4 +/- 3.6% (43.3 +/- 3.5 for males and 39.7 +/- 2.7% for females) (p < 0.001). Thirty-five percent of males and 26% of females had mild-moderate hypertension with an hematocrit of 42.3 +/- 3.5% versus 40.8 +/- 3.6% (p < 0.05). The LVMI was 132 +/- 37 g/m2 (139 +/- 38 g/m2 for males and 127 +/- 36 g/m2 for females). A significant linear association was observed between hematocrit value and diastolic BP: casual (r = 0.25; p < 0.05, 24 h mean (r = 0.34; p < 0.001), mean from 7 to 23 hours (r = 0.32; p < 0.001), mean from 23 to 7 hours (r = 0.37; p < 0.001). For females, these relationships were: r = 0.41, 0.36; 0.34 and 0.41, respectively (p < 0.01). No significant association was observed between these parameters among males. Among hypertensive patients this association was observed 24-h mean diastolic BP and mean from 23 and 7 hours (r = 0.39; p < 0.05). Among normotensive patients the values for this association were: r = 0.25; p < 0.05 and r = 0.26; p < 0.05, respectively. No significant association was observed between hematocrit value and LVMI for the total group and for the established subgroups. CONCLUSIONS: Among patients older than 55 years a weak significant association was found between hematocrit value and diastolic BP (both casual and in that obtained with ambulatory monitoring), but not with the left ventricular mass. The mean hematocrit value was significantly higher for hypertensive than for normotensive patients. PMID- 10364792 TI - [The incidence, trend and survival in lung cancer by histological type in Gipuzkoa )1983-1992)]. AB - INTRODUCTION: The incidence, trend and survival of lung cancer according to histologic types in Gipuzkoa area were studied. PATIENTS AND METHODS: The incidence of cases of lung cancer from 1983 and 1992 were studied. Cancers were classified into four major histologic groups. RESULTS: A total of 1,815 tumors were diagnosed, 91.3% carcinomas, 0.4% non-epithelial tumors and 8.3% non classified tumors. Among males, 45.5% were squamous carcinomas and among females, 49.7% adenocarcinomas. The male/female ratio of tumors has changed from 17.1/1 for the 1983-1984 period to 9.61/1 for the 1991-1992 period. Among males, a significant increase of small cell carcinomas, adenocarcinomas and large cell carcinomas was observed. Among females, a significant increase of adenocarcinomas was observed. Survival at 5 years ranged from 25.6% for the large cell carcinoma to 11.6% for the small cell carcinoma. CONCLUSIONS: Among males the squamous carcinoma and small cell carcinoma predominated; among females, adenocarcinoma was the most common type. An increase of lung cancer in both genders was observed and for all histologic types, with the exception of squamous carcinoma in males, with a plateau since 1988. The prognosis is very poor, and worse for the small cell carcinoma. PMID- 10364793 TI - [Tissue plasminogen activator as a prognostic factor in myocardial infarct patients]. AB - Changes in hemostasis inducing hypercoagulability are pathogenic factors for the development of ischemic heart disease and myocardial infarction. Nevertheless, their role is unknown in the emergence of new coronary events in patients with infarction. A group of 58 patients who had survived to a first infarction episode were studied; the cardiovascular risk factors were determined and blood concentrations of fibrinogen, t-PA, PAI and FRW measured. These patients were followed for two years to observe the development of new ischemic problems. In the study only the t-PA concentration was found to be a factor for poor prognosis. PMID- 10364794 TI - [Acute transverse myelitis in systemic lupus erythematosus]. AB - Acute transverse myelitis as complication of systemic lupus erythematosus is a known and well-characterized although uncommon clinical entity. We report here four cases of lupic myelitis collected at our hospital in the last few years and review the available literature of the last ten years, approximately the time when NMR became generally available. The clinical picture can be very variable and therefore, when facing a picture of acute myelitis, lupus should be included in the differential diagnosis; biochemistry evaluating the lupus "activity" is of poor diagnostic value, nuclear magnetic resonance is not conclusive for the etiologic diagnosis of myelitis and its prognosis has improved with therapy including pulses of steroid and immunosuppressant agents. PMID- 10364795 TI - [Idiopathic retroperitoneal fibrosis]. PMID- 10364796 TI - [Amiodarone and the thyroid]. PMID- 10364797 TI - [The standardization of the criteria for the prevention of occupational postexposure to the human immunodeficiency virus]. PMID- 10364798 TI - [An 86-year-old man with a constitutional syndrome, hemoptysis, abdominal pain and renal failure]. PMID- 10364799 TI - [Headache, an oral lesion and amaurosis in a 60-year-old man]. PMID- 10364800 TI - [Traumatic pseudocyst]. PMID- 10364801 TI - [A young oligosymptomatic woman]. PMID- 10364802 TI - [Fever, adenopathies and a papulopustular lesion of the finger]. PMID- 10364803 TI - [New prognostic factors in lung cancer]. PMID- 10364805 TI - [The HELLP syndrome: a potentially severe complication of pre-eclampsia in the immediate puerperium]. PMID- 10364806 TI - [Cisapride in idiopathic megacolon-megarectum]. PMID- 10364808 TI - [Pericholangitis and primary sclerosing cholangitis]. PMID- 10364807 TI - [Anticonvulsant hypersensitivity syndrome]. PMID- 10364809 TI - [Physiopathology of unstable angina. The role of atherosclerotic plaque rupture and thrombosis]. AB - If excess influx of lipids predominates over the proliferative response, the atherosclerotic process progresses into the formation of vulnerable lesions. This type of lesions are the most clinically relevant since they are the pathogenic basis for plaque rupture and coronary thrombus formation. Plaque rupture is a mechanical event mainly determined by the fibrous cap thickness and the lipid core size. In addition, biological factors such as inflammatory infiltration may contribute to weakening and fracture of the fibrous cap. Exposure of plaque components to flowing blood following rupture is the key event to initiate thrombosis within coronary arteries. Local factors such as quantity (fissure size), quality (plaque composition) and rheology at the site of rupture, together with systemic factors inducing hypercoagulable or thombogenic states modulate thrombosis at the time of plaque rupture. The natural history of acute coronary syndromes probably mirrors that of the underlying plaque rupture and thrombus formation. Angina stabilization would correspond to resealing of a rupture, accentuation of symptoms to development of labile thrombosis, non-Q wave infarction to development of transient thrombotic occlusion, and Q-wave infarction to establishment of a persistent occlusive thrombosis. Furthermore, this natural history may be modified by vascular tone and presence of collateral circulation. PMID- 10364810 TI - [The role of inflammation in the pathogenesis and prognosis of unstable angina]. AB - Unstable angina is a clinical syndrome which results from the unstabilization of the coronary atherosclerothic plaque, leading to its ulceration or rupture and to the formation of superimposed thrombus. The mechanism underlying plaque unstabilization is a subject of intense basic research. In the last few years, new knowledge has emerged that relates inflammation in the atherosclerothic lesion with its gradual growth and development, as well as with its sudden transformation into a complicated plaque causing unstable angina or myocardial infarction. In this article we will review the evidence that links inflammation with the pathogenesis of unstable angina and its prognosis. PMID- 10364811 TI - [Insights provided by new invasive imaging techniques into the biopathology of unstable angina]. AB - Coronary angiography only portrays a silhouette of the vessel lumen. Intravascular ultrasound allows a detailed study of the atherosclerotic plaque on the vessel wall. Intravascular ultrasound is able to determine total vessel area and, therefore, constitutes the technique of choice to analyze coronary remodelling. Alternatively, coronary angioscopy provides direct visualization of the internal coronary surface. Moreover, coronary angioscopy has the unique ability to accurately recognize coronary thrombi. Accordingly, each one of these techniques provide unique and complementary information on the anatomic substrate of patients with unstable angina. In these patients, intravascular ultrasound allows the study of plaque morphology and echogenicity, providing unique insights on its histologic composition. With current technology however, the correlation between plaque characteristics, as determined by intravascular ultrasound, and the presenting clinical syndrome is still weak. Nevertheless, in many cases intravascular ultrasound is able to recognize data suggestive of lipid pools within the plaque, ruptured plaques or findings highly suggestive of coronary thrombi. Again, the diagnosis of thrombus with intravascular ultrasound is still rather limited. On the other hand, in most patients with unstable angina coronary angioscopy is able to visualize irregular yellow plaques with associated red thrombus. Conversely, such findings are uncommon in patients with stable angina. Notably, all this information is not available using angiography and pathologic specimens (atherectomy or necropsy) from patients with unstable angina tend to keep a better correlation with intravascular ultrasound and coronary angioscopy findings than with angiographic data. From the academic point of view, both intravascular ultrasound and coronary angioscopy have revolutionized our understanding of the pathophysiology of acute coronary syndromes and have questioned old paradigms. With this in mind, the information provided by angiography has been revisited and updated. However, from a pragmatic perspective, the major challenge of these new techniques still relies on the demonstration that they may be used to assist in the decision making process involved in the management of patients with unstable angina resulting, eventually, in an improved clinical outcome. PMID- 10364812 TI - [Diagnosis of unstable angina in the emergency room. The clinical value and limitations of electrocardiography and other tests]. AB - The quick and accurate diagnosis of acute coronary heart disease in the emergency department constitutes a first line medical challenge. About 5% of patients attending to the emergency department present chest pain or symptoms suggesting myocardial ischemia. A prompt diagnosis contributes to patient survival while an incorrect one can be associated to an increased risk of unfavorable outcome. On the other hand, unnecessary admissions into high technology hospital areas determine important expenses unacceptable from a cost-benefit point of view. In the present paper, the usefulness of different diagnostic tools used in emergency department (case history, electrocardiogram, nuclear perfusion imaging, echocardiography, exercise test) is reviewed, their advantages and limitations reviewed, with special emphasis on the importance of having an electrocardiogram as soon as possible, and based on the electrocardiographic findings the most appropiate management is discussed. PMID- 10364813 TI - [Validity of different classifications of unstable angina]. AB - The classification of the unstable angina syndrome has represented one of the main objectives of the cardiologists in the two last decades. The ambiguous definition of this syndrome has led to the phenomenon that numerous classifications have been achieved, based especially in the different clinical presentations of this syndrome, that are neither clearly matched with a different physiopathology nor with the prognosis. On the other hand, the validation of the majority of the classifications have been attempted through studies of selected populations with an insufficient number of patients in a syndrome with a wide spectrum of clinical presentation, pathophysiology and prognosis. On this basis, the existing classifications do not fully satisfy the scientific community, which is confirmed by the periodical appearance of new proposals. In our setting, the classifications which are most applied are those of the Spanish Society of Cardiology and Braunwald's Classification. Both offer the usefulness of their simplicity, since they only consider clinical aspects, but sustain the previously mentioned inconveniences. A more practical classification could possibly be based exclusively on physiopathological or prognostic characteristics, which allow a more adequate management of these patients. PMID- 10364814 TI - [Differences in the clinical presentation and prognosis of unstable angina and non-Q wave infarction]. AB - At present it is generally accepted that unstable angina and non-Q-wave myocardial infarction have a common underlying pathophysiology, however, clinical presentation and outcome of these syndromes depend on the location, grade and duration of myocardial ischemia. The most important difference between both syndromes is the myocardial necrosis from non-Q myocardial infarction, which may carry a higher risk death and reinfarction rate and for that reason a worse prognosis than unstable angina. Some studies suggest that the effect of antithrombotic treatment can be different in both syndromes. Nevertheless the differences do not achieve significance in most of these studies. PMID- 10364815 TI - [Current prognosis of unstable angina. The results of clinical studies]. AB - In the last decade there has been a change in the clinical evolution of patients with unstable angina. A better knowledge of his pathophysiology has improved the progress in their treatment and prognosis. Aspirin and heparin have played an important role in this change. Nowadays, the rates of death and acute myocardial infarction at 6 weeks are about 2% and 5% respectively. Nevertheless, the debate about the stratification risk and the best therapeutic options persist. Part of this debate corresponds to the classification differences and to the heterogeneity of variables considered to be end points. On the other hand, the clinical importance conferred to variables identified as predictor of worse prognosis is not valued enough, forgetting that the low prevalence of events results in a low positive predictive value. PMID- 10364816 TI - [Platelet activation and inhibition. Old and new platelet antiaggregants]. AB - Aggregates of activated platelets, at the level of a plaque fissure, seem to have a pivotal role in acute coronary syndromes. The glycoprotein IIb-IIIa receptors of the platelet surface are thought to be the final common pathway of platelet aggregation. The possibility of using a basic treatment to control platelet aggregation, in the whole clinical spectrum of acute coronary syndromes is attractive. The results of different clinical studies show evidence that glycoprotein IIb-IIIa antagonists, given intravenously, are effective in the management of acute coronary intervention with high thrombogenic risk, and also help to control the early phase of acute coronary syndromes. In patients who have responded to the acute i.v. infusion, the chronic oral administration of those agents might contribute to their clinical stabilization and reduce the risk of further coronary events. PMID- 10364817 TI - [The importance of thrombin and its inhibition in unstable angina. Nonfractionated heparin, hirudin and defractionated heparins]. AB - Atheroma plaque rupture with liberation of tissue factor activates the coagulation cascade and plateletes, leading to the formation of intracoronary thrombi in many patients with acute coronary syndromes. In this process, tissue factor, thrombin, Factor Xa and fibrin play a major role. This review analyses the clinical efficacy of the antithrombotic drugs: fractionated heparin, low molecular or fractionated heparins, direct thrombin inhibitors, specific Xa factor inhibitors and inhibitors of the tissue factor pathway in patients with unstable angina and non-Q wave myocardial infarction. Enoxaparin, a low molecular weight fractionated heparin, has shown to be associated with a greater clinical efficacy, superior to that achieved with conventional heparin anticoagulation or treatment with aspirin, and probably should be considered as the antithrombotic of choice. Present clinical research should be aimed at the identification of patients with greater benefit, new treatment protocols with other antithrombotic drugs and the efficacy in special situations such as invasive coronary interventions or the association with other drug like, thrombolytic agents or new antiplatelet antiaggregants. PMID- 10364818 TI - [Persistent activation and/or reactivation in the subacute phase of unstable angina. Reasons for prolonged antithrombotic treatment]. AB - Coronary artery thrombosis, due to a rupture of the vulnerable plaque, plays an important role in acute coronary syndromes. The interaction between platelets and thrombin with ruptured vulnerable plaque trigger a complex mechanism. The clinical manifestations depend on the extent and duration of thrombus formation. In the acute phase, aspirin, heparin and the new drugs reduce ischemic clinical outcomes. However, clinical rebound after withdrawal antithrombotic therapy has been observed and, follow-up studies have also documented a high risk of recurrence in the following months. A hypercoagulable state, thrombin generation and activation and haematological rebound is shown in acute coronary syndrome patients. Thus, the goal of treatment could be to control thrombotic response in the acute phase and to allow the healing and stabilization of the culprit lesion to avoid clinical ischemic events in the long-term. PMID- 10364819 TI - [Identification of patients at high risk in the initial evaluation of unstable angina. Clinical importance of electrocardiogram, Holter and biochemical markers in myocardial lesions]. AB - Beneath the expression of unstable angina are joined an heterogeneous group of patients with different prognosis. In the initial presentation it is difficult to differentiate from non-Q wave myocardial infarction. The present review attempts to establish the relative importance of the clinical presentation, the electrocardiogram, the Holter monitoring and the biochemical markers of myocardial damage for risk stratification of these patients. The clinical factors related with prognosis are the general risk factors related to the prognosis in other presentations of the ischemic heart disease as the presence of previous myocardial infarction, age, presence of heart failure or left ventricular dysfunction, diabetes and peripheral vascular disease. The electrocardiogram is the most helpful method for risk stratification of unstable angina patients, especially depression. The continuous monitoring of the ST-segment through Holter monitoring constitutes an important investigation area, but presumably it can not be recommended as routine utilisation yet in the managing of this sort of patients. The rising of cardiac troponins T or I constitute other important risk marker. Possibly three different risk groups could be established: a) patients at low risk, those with normal electrocardiogram or only negative T-waves and normal cardiac troponins; b) intermediate risk, those patients with rising of cardiac troponins and normal electrocardiogram or electrocardiogram with ST-segment depression and normal cardiac troponins, and c) high risk, corresponding to the patients with rising of cardiac troponins and electrocardiogram with ST-segment depression. PMID- 10364820 TI - [Noninvasive risk stratification of patients with unstable angina in the acute phase. The prognostic value of exercise and stress after drug treatment]. AB - Most of the prognostic information on patients with unstable angina is obtained from the initial clinical assessment and the patient's subsequent course over the first few days of management. In recently stabilized patients noninvasive stress testing with exercise or pharmacologic testing provide additional useful risk assessment. Noninvasive stress testing should be part of the outpatient evaluation of low-risk patients with unstable angina and should be done within 72 hours. Unless cardiac catheterization is indicated, noninvasive stress testing should be performed in low- or intermediate-risk patients hospitalized with unstable angina who have been free of symptoms for a minimum of 48 hours. Choice of stress testing modality should be based on an evaluation of the patient's resting ECG, his or her physical ability to perform exercise, and the local expertise and technologies available. The standard ECG treadmill test should be the standard test employed in most patients with no contraindications for doing so. Provocation of ischemia at a low workload (< 5 to 6 METs) signifies a high risk patient (cardiac mortality > or = 4%/year) who should generally merit referral to cardiac catheterization. Attainment of a higher workload (> 5 to 6 METs) without ischemia is associated with a better prognosis (cardiac mortality < 1%/year), and many such patients can be managed conservatively. Those who tolerate only a low workload but have no evident ischemia or those who develop ischemia at a high workload, represent an intermediate-risk group (cardiac mortality 2 to 3%/year) for whom several reasonable strategies can be proposed. PMID- 10364821 TI - [Early percutaneous revascularization in patients with unstable angina. Current results and comparison with conservative medical treatment]. AB - In patients with ongoing angina, despite optimal medical therapy, the best therapeutic alternative is coronary angiography followed by emergency coronary revascularization with surgery or angioplasty. However, whether or not all patients should have early angiography and revascularization is a matter of debate. This paper reviews the advantages of modern medical therapy in this setting and the problems associated with early coronary revascularization. In particular, it analyses the data from the main clinical trials that have specifically compared an early invasive procedure with a conservative strategy in unstable coronary syndromes. Finally, it assesses the impact of the new antithrombotic agents, such as glycoprotein IIb/IIIa receptor blockers, particularly during coronary percutaneous interventions. The data reviewed suggest that early invasive intervention should be reconsidered, and that patients should be controlled (if possible) under medical treatment until non invasive stratification tests allow the identification of those patients who would benefit most from revascularization. PMID- 10364822 TI - [Angioplasty for acute coronary syndromes in the era of the stent]. AB - Pharmacological treatment frequently stabilizes symptoms of patients with acute myocardial ischemia. However, significant quiescent residual stenosis normally persists and leads to rethrombosis. Since rethrombosis produces reischemia and has a deleterious impact on initial and long-term prognosis in these patients, definitive normalisation of local flow assured through deactivation treatments and complete passivation of quiescent residual stenosis in an inert plaque should be a main priority in modern treatment of acute coronary ischemia. Considering the negative influence of significant stenosis on rethrombosis, and that the normalization has a clear antithrombotic effect of flow, routine elimination of residual stenosis by means of angioplasty should prevent rethrombosis and its side effects. Nevertheless, according to trials carried out previous to the most relevant progresses in the field of interventional cardiology, the advantage of this strategy over the conservative treatment has not been clearly demonstrated. Coronary stenting produces a real normalisation of flow and lumen which prevents local thrombosis. In concordance with these facts, recent evidence indicates a substancial clinical benefit of stenting in very thrombogenic acute settings, such as primary angioplasty or refractory acute coronary angina. Presumably, routine stenting also benefits initial and long-term prognosis of other subsets of unstable patients, especially those with thrombolysed myocardial infarction and stabilized patients with acute ischemia without ST-segment elevation. To demonstrate this new trials are needed to compare the efficacy of conservative and interventional approaches that incorporate the advances of each strategys. Until new data are available on these studies, the treatment of acute coronary ischemia should be tailored to each patient and no out-dated recommendation should be given or accepted. PMID- 10364823 TI - [The role cardiac surgery in the myocardial revascularization of patients with unstable angina in the era of interventional cardiology]. AB - Indications for surgical myocardial revascularization in patients with unstable angina have been extended during the last years achieving similar results to those obtained in stable angina. On the other hand, percutaneous transluminal coronary angioplasty in this same subgroup of patients have achieved better results specially after the advent of intracoronary stenting. Selection of a specific technique of revascularization may be nowadays difficult and the update indications for both surgery and angioplasty are discussed. PMID- 10364824 TI - Variability and evolvability of male song characters in Drosophila montana populations. AB - In Drosophila montana, the male courtship song (especially the pulse characters of the song) plays an important role in sexual selection. The heritabilities and the amount of additive and residual variation in different characters of the male song were measured in two populations using father-son regression and sib analysis. The songs of the males from the Oulanka population were recorded for a second time after the males had been kept in 4 degrees C for 6 months. Heritabilities measured for different song traits were non-significant in each case, largely due to high residual variation. During the cold treatment, the additive variation increased and the residual variation decreased in nearly all song traits. Our results suggest that genotype-environment interactions may increase the amount of additive variation between males in sexually selected song traits during overwintering, prior to the mating season of the flies. PMID- 10364825 TI - Characterization of the lysinuric protein intolerance (LPI) region within T-cell receptor alpha/delta gene cluster on chromosome site 14q11. AB - Lysinuric protein intolerance is a recessively inherited metabolic disease characterized by defective efflux of cationic amino acids at the basolateral membrane of the intestinal and renal tubular epithelium. Linkage analysis and further linkage disequilibrium in Finnish LPI families have earlier assigned LPI gene locus within or in close vicinity of T-cell receptor alpha/delta gene cluster on chromosome site 14q11. In the present work we have characterized the linkage defined LPI region using RH-mapping and fiber-FISH and searched the LPI gene from the reported sequence of the T-cell receptor gene. PMID- 10364826 TI - Geographic variation of six allozyme loci in Drosophila melanogaster: an analysis of data from different continents. AB - Based on data from 70 literature sources, the frequencies of common alleles at six allozyme loci were examined in Drosophila melanogaster populations from five geographic regions: North America (including Central America), South America, Europe-Africa, Asia, and Australasia. The analyzed loci were Adh, Odh, Gpdh, G6pd, Pgd, and Est-6, which have been previously reported by other authors to show latitudinal variation in North America, Eurasia and Australasia. We found five parallel latitudinal clines for AdhF and three clines for GpdhS in five geographic regions as well as four clines for G6pdF, three clines for Est-6S, and two clines for OdhF and PgdF in four regions (data from South America for G6pd, Odh, Est-6, and Pgd were not available). Such pattern of variation confirmed the possibility that considered allozyme polymorphisms are maintained by climatic selection. Significant differentiation of mean allele frequencies among geographic regions was in agreement with current evidence on history of D. melanogaster worldwide dispersion. PMID- 10364827 TI - Genetic characterization of two Eragrostis species using AFLP and morphological traits. AB - Tef [Eragrostis tef (Zucc.) Trotter] is the most important cereal crop in Ethiopia. An experiment was conducted to investigate genetic diversity among four cultivars of tef and 14 accessions of Eragrostis pilosa using radiolabelled and silver stained amplified fragment length polymorphism. Morphological traits were also evaluated. A total of 897 markers were obtained out of which 395 were polymorphic using 11 primer combinations. Cluster analysis revealed accessions of E. pilosa which are distantly related and others closely related to tef. Our previous experience also indicates that E. pilosa is crossable with tef. Those accessions distantly related to tef could be used in a crossing program to generate a population for selection and/or genetic mapping. Such genetic mapping populations will form an important entry point towards the molecular genetic dissection of the plant genus, Eragrostis, especially in the context of comparative mapping. Knowledge gained from such study, apart from tef improvement, will also be useful for many forage and turf grass species where little molecular genetic information is available. Nine cultivars or accessions had one or more unique fragments using one or more AFLP primers indicating the potential of the technology in fingerprinting tef in a breeding or seed multiplication program. The results also showed that clusters obtained using silver staining and gamma 33P-ATP labeling were similar, suggesting that silver staining could be used as an alternative to radiolabeling at least in genetic diversity analysis. Significant genetic variation was obtained for morphological traits. Of particular interest to tef breeding was short plant stature in E. pilosa which could be transferred to tef to minimize the problem of lodging. Diversity revealed at the morphological trait level was not commensurate with that observed for AFLP. This was due to the small number of available morphological traits and their interaction with the environment. PMID- 10364828 TI - RAPD mapping in a doubled haploid population of rice (Oryza sativa L.). AB - To examine the distribution and genome coverage of RAPDs, a total of 242 Random Amplified Polymorphic DNA (RAPD) markers generated by 73 random decamer primers were mapped onto 12 rice chromosomes by linkage analysis using a doubled haploid population, developed from an indica x japonica cross. The RAPD markers were derived from both parents equally and were well distributed over the rice genome. Furthermore, multiple RAPD markers generated from the same primer were dispersed over different chromosomes rather than clustered. The RAPD technique provided improved marker coverage on a previously developed RFLP map. A set of primers producing reproducible markers originating from either parent and equally spaced over all the 12 chromosomes were selected for application in marker-assisted backcross breeding. The RAPD analysis as a realistic and practical alternative to RFLP and their usefulness in anchoring the identified BAC contigs directly to chromosomes is discussed. PMID- 10364829 TI - The acid phosphatase-1 gene region in the Drosophila species of the subobscura cluster. AB - The acid phosphatase-1 (Acph-1) gene region was sequenced in three species of Drosophila: D. subobscura, D. madeirensis and D. guanche. These three closely related species, which are included in the obscura group, form the subobscura cluster. The different functional regions of the gene were identified by similarity with the sequence of D. melanogaster. The structure of Acph-1 is conserved in the four species. Average divergence at synonymous and nonsynonymous sites between D. melanogaster and the species of the subobscura cluster is Ks = 1.1354 and Ka = 0.1743, respectively. The rather high Ka value confirms that ACPH 1 is a rapidly evolving enzyme in Drosophila, as previously suggested by immunological studies. Amino acid replacements are not randomly distributed along the gene. In fact, an excess of replacements is detected in exon I, indicating that the signal peptide encoded by this exon evolves even faster than the rest of the protein. Divergence at the Acph-1 gene region is further evidence that D. madeirensis and D. subobscura are more closely related than D. guanche is to any of them. In addition, both silent divergence in noncoding regions and synonymous divergence in the coding region indicate that the split of the D. guanche lineage is about twice as old as the split of the lineages leading to D. madeirensis and D. subobscura. These phylogenetic relationships are, however, not supported by divergence at nonsynonymous sites since the lowest Ka estimate is between D. guanche and D. subobscura. PMID- 10364830 TI - A nucleotide sequence linked to the vrs1 locus for studies of differentiation in cultivated barley (Hordeum vulgare L.). AB - A PCR-amplified DNA, cMWG699, is linked to the vrs1 (formerly v) locus controlling 2- and 6-rowed spikelets. Restriction analysis of the amplified DNA of 65 varieties from Europe, America, and East Asia revealed 3 alleles, named types K, A and D. Two-rowed varieties were mostly of type K allele, and 6-rowed varieties were mostly of type A allele. The type D allele was found only in three 6-rowed varieties. Sequence comparison of these alleles revealed that the type A allele is more closely related to the type K allele than to the type D allele. The sequence analysis also suggested that the types A and D alleles diverged earlier than when 6-rowed barley arose. On the assumption that 2-rowed barleys were the ancestors of 6-rowed barley, 6-rowed barleys with types A and D alleles probably differentiated from 2-rowed barleys with type A and D alleles, respectively, by independent mutations on the vrs1 locus. PMID- 10364831 TI - Hypericin as a non-antioxidant inhibitor of NF-kappa B. AB - NF-kappa B is a transcription factor involved in immune and inflammatory responses. Here we show that micromolar concentrations of hypericin inhibited the PMA- and TNF-alpha-induced activation of NF-kappa B in HeLa and TC10 cells, respectively. In contrast, NF-kappa B activated by H2O2 was not influenced by hypericin, indicating a pathway-specificity of hypericin. Hyperforin and a Hypericum perforatum extract standardised on 0.15% hypericin and 5% hyperforin were not active at pharmacologically relevant concentrations. The PMA/TNF-alpha induced transcription of a reporter gene, which is under the control of the NF kappa B-dependent IL-6 promoter, was strongly reduced by preincubation with hypericin. PMID- 10364832 TI - Heparinoids: structure, biological activities and therapeutic applications. AB - Heparin is an important polyanionic drug having a wide variety of different biological activities. Substantial research effort has focused on the preparation of improved heparins and heparin analogues that might exhibit higher specificity and decreased side effects. These heparin analogues or heparinoids include sulfated polysaccharides from plant and animal origin, synthetic derivatives of polysaccharides, and acidic oligosaccharides and their small synthetic analogues. The structure, biological activities and therapeutic potential of these heparinoids are discussed. PMID- 10364833 TI - Reversal of multidrug resistance by kopsiflorine isolated from Kopsia dasyrachis. AB - Kopsiflorine, an indole alkaloid of the aspidofractinine-type isolated from Kopsia dasyrachis, was examined for its effect in enhancing drug cytotoxicity in multidrug-resistant tumor cells. The cytotoxicity of vincristine was enhanced in a concentration-dependent manner by kopsiflorine in drug-resistant KB cells (VJ 300). Kopsiflorine alone had no effect on the growth of drug sensitive or resistant cells, but the intracellular accumulation of vincristine was enhanced by kopsiflorine in VJ-300 cells. Kopsiflorine (10 micrograms/ml) significantly inhibited the binding of [3H]azidopine to P-glycoprotein in VJ-300 cells. The results suggest that kopsiflorine interacts directly with P-glycoprotein and inhibits the efflux of antitumor agents in drug-resistant cells. PMID- 10364834 TI - Cytotoxic activity of indole alkaloids from Alstonia macrophylla. AB - Thirteen indole alkaloids isolated from the root bark of Alstonia macrophylla and a semisynthetic bisindole O-acetylmacralstonine have been assessed for cytotoxic activity against two human lung cancer cell lines, MOR-P (adenocarcinoma) and COR L23 (large cell carcinoma), using the SRB assay. Pronounced cytotoxic activity was exhibited by the bisindoles on both cell lines. This suggests that, in comparison with the corresponding monomeric indoles, at least part of both the ring systems present in the bisindoles is essential for cytotoxic activity. The potent alkaloids were further tested against a human normal cell line (breast fibroblasts) and other human cancer cell lines including StMI1 1a (melanoma), Caki-2 (renal cell carcinoma), MCF7 (breast adenocarcinoma), and LS174T (colon adenocarcinoma). The bisindoles O-acetylmacralstonine, villalstonine and macrocarpamine were found to possess pronounced activity against cancer cell lines with IC50 values in the range of 2-10 microM, with no discernible cell-type selectivity. However, O-acetylmacralstonine displayed discernibly less toxicity against the normal breast fibroblasts. PMID- 10364835 TI - In vitro leishmanicidal activity of aurones. AB - A series of aurones with drug-potential for Leishmania infections was identified in vitro using both a direct cytotoxicity assay against extracellular promastigotes of Leishmania donovani, L. infantum, L. enriettii, and L. major, and a test against intracellular amastigote forms of L. donovani residing within murine macrophages. The most active aurone (6-hydroxy-2-[phenylmethylene]-3(2H) benzofuranone) had an EC50 of 0.45 microgram/ml in the extra-, and an EC50 of 1.40 micrograms/ml in the intracellular assay. Other aurones were active between 0.06-12.50 micrograms/ml and 0.04-7.81 micrograms/ml, respectively. When tested against murine bone marrow-derived macrophages as a mammalian host cell control, the compounds showed only moderate cytotoxicity (EC50 2.32 to > 25.0 micrograms/ml). This is the first report on aurones as a new class of natural products with leishmanicidal activity. PMID- 10364836 TI - Effect of several sesquiterpene lactones on lipid peroxidation and glutathione level. AB - Seven sesquiterpene lactones isolated from Helenium aromaticum: helenalin, mexicanin I, linifolin A, geigerinin, and from Telekia speciosa: 6 alpha-hydroxy 2,3-dihydroaromaticin, asperilin, telekin have been tested for their hydroxyl radical scavenging activity and effect on lipid peroxidation. All compounds were found to be potent hydroxyl radical scavengers but did not affect lipid peroxidation in vitro. In vivo they exerted pro-oxidative properties and caused glutathione level depletion and elevation in glutathione peroxidase activity. PMID- 10364837 TI - Antiulcerogenic mechanisms of dehydrocrotonin, a diterpene lactone obtained from Croton cajucara. AB - The bark of Croton cajucara Benth, is used in Brazilian folk medicine as an infusion to treat gastrointestinal disorders. The aim of the present study was to assess the mechanisms involved in the antiulcerogenic activity of dehydrocrotonin (DHC), a diterpene isolated from C. cajucara bark. We studied the effects of DHC on pylorus ligature (Shay) in mice treated with the drug (100 mg/kg) by the intraduodenal route. DHC did not induce any alteration in gastric volume in Shay mice but modified the pH and total acid concentration of gastric juice. Incubation of gastric juice with DHC did not reduce gastric acidity compared to control. We also investigated the effects of DHC on the response to histamine of right atria isolated from guinea pigs and on the response to carbachol of stomach fundus strips from rats. The concentration-response curves for the chronotropic effect of histamine in guinea pig right atria were shifted to the right, with a significant decrease in the maximum response, in the presence of DHC. Similar results were obtained with DHC (30 microM) for the concentration-response curves to carbachol in the isolated rat stomach. The ability of DHC to increase PGE2 release from rat stomach mucous cells was also studied. We observed that DHC induced a significant increase in PGE2 production (60% compared to control). In addition, the effects of DHC on the healing of acetic acid-induced gastric ulcer in rats were evaluated 14 days after acid injection. Oral administration of DHC (100 mg/kg per day) for 14 consecutive days had no effect on gastric ulcer healing in rats. Thus, the protective effect of DHC on induced gastric lesions could be due to synergistic effects, e.g., an increase in PGE2 release and non competitive antagonism of H2-receptors and of muscarinic receptors. Whereas the former result represents an increase in the protective factors, the latter one shows a decrease in the aggressive factors against the gastric mucosa. PMID- 10364839 TI - Crataegus extract blocks potassium currents in guinea pig ventricular cardiac myocytes. AB - Crataegus extract is used in cardiology for the treatment of mild to moderate heart failure (NYHA II) in Germany. However, little is known about the electrophysiological actions of Crataegus extract in the heart. Recently, it was shown that Crataegus extract prolongs the refractory period in isolated perfused hearts and increases action potential duration in guinea pig papillary muscle. It was the aim of this study to find out the mechanism of the increase in action potential duration caused by Crataegus extract. Using the patch-clamp technique, we measured the effects of Crataegus extract (10 mg/l; flavonoid content: 2.25%, total procyanidin content: 11.3 +/- 0.4%) on the inward rectifier and the delayed rectifier potassium current in isolated guinea pig ventricular myocytes. To get some insight into the mechanism underlying the positive inotropic effect of Crataegus extract, we also looked for effects on the L-type calcium current. Crataegus extract slightly blocked both the delayed and the inward rectifier potassium current. The inhibition amounted to 25% and about 15%, respectively. This amount of inhibition of these repolarising currents is sufficient to explain the prolongation of action potential duration caused by Crataegus extract. To our surprise we could not detect any influence of Crataegus extract on the L-type calcium current. In summary, our results show that Crataegus extract blocks repolarising potassium currents in ventricular myocytes. This effect is similar to the action of class III antiarrhythmic drugs and might be the basis of the antiarrhythmic effects described for Crataegus extract. Our measurements of the L type calcium current indicate that Crataegus extract's positive inotropic effect is not caused by phosphodiesterase inhibition or a beta-sympathomimetic effect. PMID- 10364838 TI - Hypotensive, hypoglycaemic and toxicological studies on the flavonol C-glycoside shamimin from Bombax ceiba. AB - Shamimin, a C-flavonol glucoside from Bombax ceiba leaves showed significant potency as a hypotensive agent at the doses of 15 mg/kg, 3 mg/kg, 1 mg/kg and significant hypoglycaemic activity at 500 mg/kg in Sprague-Dawley rats. Further studies revealed that it did not cause any mortality in mice at the dose of 1 g/kg but in rats 500 mg/kg is a lethal dose. Aqueous and methanolic extracts of Bombax ceiba leaves and one of its fractions were also subjected to pharmacological and toxicological screening. PMID- 10364840 TI - Calcium influx inhibition: possible mechanism of the negative effect of tetrahydropalmatine on left ventricular pressure in isolated rat heart. AB - The active ingredient dl-tetrahydropalmatine (THP) isolated from the traditional Chinese herb Corydalis racemosa has been found to have antihypertensive effects. However, severe cardiac and neurological toxic effects were reported from using this herb for the treatment of pain. In an isolated perfused rat heart model, THP at the concentration of 100 microM was found to have a negative effect (-45%) on left ventricular pressure and this effect was produced concentration-dependently from concentrations lower than 50 microM. In isolated cardiomyocytes, radioactive calcium influx was also inhibited significantly by THP at the concentration of 100 microM and this effect was also in a concentration-dependent manner (-39%). In a patient with latent heart disease, the use of Corydalis should probably be detrimental, the toxic effect was probably due to calcium influx inhibition. PMID- 10364841 TI - Biological and chemical characterization of the fraction with antiherpetic activity from Achyrocline flaccida. AB - The in vitro antiviral activity demonstrated by aqueous extract (AE) of Achyrocline flaccida on herpes simplex virus type-1 is exerted early during the viral replication, essentially during the viral adsorption to host cells. A bioguided purification process of the AE indicated that negatively charged polysaccharides were responsible for this antiviral activity. PMID- 10364842 TI - Antimycobacterial eudesmanolides from Inula helenium and Rudbeckia subtomentosa. AB - In a bioassay guided search for antimycobacterial compounds from higher plants, the root extracts of Elecampane (Inula helenium L.; Asteraceae) and Sweet Coneflower (Rudbeckia subtomentosa Pursh.; Asteraceae) were chemically investigated for their active constituents. Chromatographic fractions of root extracts of l. helenium, which exhibited significant activity against Mycobacterium tuberculosis, provided the known eudesmanolides alantolactone, isoalantolactone, and 11 alpha H, 13-dihydroisoalantolactone. Peracid epoxidation of alantolactone and isoalantolactone provided 5 alpha-epoxyalantolactone and 4(15) alpha-epoxyisoalantolactone, respectively and oxidation of alantolactone with OsO4 gave 11,13-dihydroxyalantolactone. Active fractions from R subtomentosa contained the known alloalantolactone and 3-oxoalloalantolactone. The structures of the above compounds were established by spectroscopic methods including 1D and 2D NMR techniques as well as spectral comparison with previously reported data. The molecular structure of 5 alpha-epoxyalantolactone was determined by single crystal X-ray diffraction. Eleven natural and semisynthetic eudesmanolides were tested in a radiorespirometric bioassay for activity against M. tuberculosis. 5 alpha-Epoxyalantolactone and encelin from Montanoa speciosa showed minimum inhibitory concentrations (MICs) of 8 and 16 micrograms ml-1, respectively. Alantolactone, isoalantolactone and its 4 alpha, 15-epoxide, 1,2-dehydro-3-epi isotelekin and alloalantolactone gave MICs of 32 micrograms ml-1. All other compounds showed MIC values of 128 micrograms ml-1 or higher. PMID- 10364843 TI - Three novel bicyclic taxane diterpenoids with verticillene skeleton from the needles of Chinese yew, Taxus chinensis var. mairei. AB - Three novel bicyclic taxane diterpenoids with the verticillene skeleton were isolated from the needles of Chinese yew, Taxus chinensis var. mairei. Their structures were established as (3E,7E)-2 alpha, 10 beta, 13 alpha, 20 tetraacetoxy-5 alpha-hydroxy-3,8-secotaxa-3,7,11-trien-9-one (1), (3E,7E)-2 alpha, 5 alpha, 10 beta, 13 alpha-tetraacetoxy-20-hydroxy-3,8-secotaxa-3,7,11 trien-9-one (2), and (3E,7E)-10 beta, 13 alpha-diacetoxy-2 alpha, 5 alpha, 20 trihydroxy-3,8-secotaxa-3,7,11-trien-9-one (3) on the basis of 1D, 2D NMR and HR MS spectroscopic analysis. Verticillene skeleton was considered as the biogenetic intermediate of taxane diterpenoids, isolation of bicyclic taxane diterpenoids with verticillene skeleton from this plant provides direct proof for this hypothesis. PMID- 10364844 TI - Molecular diversity of 5S-rRNA spacer domain in Fritillaria species revealed by PCR analysis. AB - Beimu (bulbs of Fritillaria) is an important traditional Chinese herbal medicine commonly used as an antitussive and expectorant. There are about 25 species and varieties of Fritillaria that carry the name Beimu on commercial markets. The price for each Beimu may differ by more than 100-fold. However, the identification of the origin of a particular species on the market is difficult. Here, a molecular method was used to identify various species of Fritillaria regardless of their geographical origin. The 5S-rRNA coding sequence is highly conserved in higher eukaryotes, but the spacer sequence of the 5S-rRNA gene is variable among different species. Total genomic DNA from fresh leaves and bulbs of Fritillaria cirrhosa, F. puqiensis, F. anhuiensis, and F. thunbergii was extracted. The 5S-rRNA spacer region of the extracted DNAs was amplified by PCR with a pair of primers located within the conserved coding region. The isolated cDNA clones (approximately 600 bp) covering the 5S-rRNA spacer domain were sequenced. By aligning the isolated nucleotide sequences of the four Fritillaria species, sequence diversity was found in the spacer region. Furthermore, a unique EcoR I site was used for the rapid identification of different species of Fritillaria. To our knowledge, this is the first report on the detection of 5S rRNA spacer domain sequences of Fritillaria and their use to identify species. PMID- 10364845 TI - Colony stimulating factor-inducing activity of hesperidin. AB - To evaluate immunomodulating activities of bioflavones, colony-stimulating factor (CSF)-inducing activity of two dihydroflavones, three flavones and three flavonols were tested. These samples were suspended in saline and injected intraperitoneally (i.p.) into mice at a dose of 1 mg 6 h before bleeding. All compounds carrying the glucosyl-rhamnose moiety showed potent activity. Among them, hesperidin exhibited the strongest activity. Serum CSF production in mice injected with 1 mg hesperidin reached a peak at 9 h later. The activity of hesperidin was dose-dependent at a range of 0.3 to 20 mg/mouse. PMID- 10364846 TI - The anti-inflammatory compounds of Polygonum bistorta: isolation and characterisation. AB - A bioassay-guided technique has been used to isolate anti-inflammatory compounds from the dried rhizomes of Polygonum bistorta, for structural identification. Anti-inflammatory activity was detected using the carrageenan-induced rat paw oedema. Two compounds were isolated which significantly suppressed the inflammatory response. Spectral data from EIMS and NMR together with some physical characteristics revealed the identities of the two active compounds as 5 glutinen-3-one and friedelanol. The results indicate that these two compounds largely account for the anti-inflammatory actions of the rhizomes of P. bistorta. PMID- 10364847 TI - Inhibitory effect of triterpenes from Crataegus pinatifida on HIV-I protease. AB - The methanol extracts of the leaves of Crataegus pinnatifida showed potent inhibitory activities against HIV-1 protease at a concentration of 100 micrograms/ml. The subsequent fractionation and isolation of the extract gave two active compounds. Their structures were identified as uvaol (1) and ursolic acid (2) by spectral data. These active compounds inhibit HIV-1 protease with IC50 values of 5.5 and 8.0 microM, respectively. PMID- 10364848 TI - Cytotoxic 4-nerolidylcatechol from Pothomorphe peltata inhibits topoisomerase I activity. AB - Bioactivity-guided fractionation of the leaf methanolic extract of P. peltata (Piperaceae), using the brine shrimp lethality test, led to the isolation of catechol derivative 4-nerolidylcatechol (4-NC). The methanolic extract was active against crown gall tumour in potato discs, showing a 22% crown gall tumour inhibition (SD = 4%), while 4-NC was cytotoxic against KB tumour cells growth (EC50 = 1.3 micrograms/ml). No interaction with DNA could be observed when tested using the methyl green-DNA (MG-DNA) bioassay. An inhibition in the activity of topoisomerase I using agarose gel electrophoresis was detected in the presence of the purified compound (IC50 = 20 micrograms/ml), suggesting that this could be a possible mechanism for the cytotoxicity observed in KB cells. PMID- 10364849 TI - In vitro antimalarial activity and cytotoxicity of cochlospermum tinctorium and C. planchonii leaf extracts and essential oils. AB - The antimalarial and toxicological properties of Cochlospermum tinctorium and C. planchonii extracts and essential oils prepared from their leaves were studied. The oil components were extracted by hydrodistillation of the plant leaves and characterized by gas chromatography and mass spectrometry. Crude extracts and oils were tested for in vitro antimalarial activity on Plasmodium falciparum. The IC50 were evaluated after 24 and 72 h contact between the oils and the parasite culture, and ranged from 22 to 500 micrograms/ml. C. planchonii leaf oil yielded the best antimalarial effect (IC50: 22-35 micrograms/ml), while the most potent effect from crude leaf extracts was induced by C. tinctorium. The cytotoxicity of the leaf crude extracts and oils was assessed on the K562 cell line and showed IC50 values ranging between 33 and 2000 micrograms/ml. PMID- 10364850 TI - Inhibition of activator protein 1 activity by berberine in human hepatoma cells. AB - Activator protein 1 (AP-1) is a transcription factor which plays a critical role in inflammation and carcinogenesis. The present study was conducted to investigate the effect of berberine, an isoquinoline alkaloid present in plants of the genera Berberis and Coptis, on the activity of AP-1 using a reporter gene assay in human hepatoma cells. Berberine was shown to inhibit AP-1 activity in a dose- and time-dependent manner at concentrations higher than 0.3 microM. Berberine inhibited AP-1 activity almost completely as low as 10 microM after 48 h treatment. The inhibitory effect on AP-1 activity in cancer cells may further explain the anti-tumor promoting activity of berberine. PMID- 10364851 TI - Shamimin: a new flavonol C-glycoside from leaves of Bombax ceiba. AB - Shamimin a new flavonol C-glycoside has been isolated as a pale yellow powder from the ethanolic extract of fresh, undried leaves of Bombax ceiba. Its structure has been elucidated as 2-(2,4,5-trihydroxyphenyl)-3,5,7-trihydroxy-6-C- glucopyranosyloxy-4H-1-benzopyran-4-one through extensive spectroscopic methods (IR, mass, 1H- and 13C-NMR), and 2D-NMR experiments. Shamimin showed antimicrobial activity against a few bacteria and fungi. PMID- 10364852 TI - Hinokiresinol: a novel inhibitor of LTB4 binding to the human neutrophils. PMID- 10364853 TI - Why mothers die--report on confidential enquiries into maternal deaths in the United Kingdom 1994-96. PMID- 10364854 TI - Cortical responses to auditory stimuli during isoflurane burst suppression anaesthesia. AB - The cortical responses to auditory stimuli were studied in 12 patients during isoflurane anaesthesia producing burst suppression (ETisof (SD) 1.4 (0.2) vol.%). Earphones were used to give 3-s trains of auditory click stimuli (60 clicks, 20 clicks per second, 80 dB, 0.1 ms) at irregular intervals. In 10 patients, the electroencephalography (EEG) showed a burst suppression pattern consisting of high-amplitude activity intermingled with suppressed background activity. In eight patients with burst suppression patterns, there was a strong cortical reactivity to the termination, not to the beginning, of auditory stimuli: 80 (20)% of all stimuli presented during EEG suppression evoked high amplitude cortical response, offset-burst. The latency of these auditory offset evoked bursts was 540 (60) ms. Auditory offset evoked bursts suggest that in spite of cortical suppression during deep anaesthesia the brain retains its ability to respond to changes in the acoustic environment. PMID- 10364855 TI - Comparison of supine and upright positions on autonomic nervous activity in late pregnancy: the role of aortocaval compression. AB - In order to understand the effect of aortocaval compression on autonomic nervous activity, we compared the effects of supine and upright postures on measures of heart rate variability in late pregnancy. Thirty-two women in late pregnancy and 23 nonpregnant age-matched women were studied. Both time and frequency domain heart rate variability measures were compared between the supine and upright positions, and between pregnant women and controls. The pregnant women had smaller mean RR interval, standard deviation of RR intervals and normalised high frequency power, and a larger low- to high-frequency power ratio than nonpregnant women in the supine position. When the position was changed from upright to supine in nonpregnant women, the percentage change in the mean RR interval and normalised high-frequency power were increased, whereas the percentage change in the coefficient of variation of RR interval and low- to high-frequency power ratio were decreased. In contrast, the percentage change in normalised high frequency power was decreased and the percentage change in low- to high-frequency power ratio was increased in pregnant women when the position was changed from upright to supine. These results indicate that the vagal enhancing effect seen in normal subjects when changing position from upright to supine was reversed in women in late pregnancy. Aortocaval compression in the supine position in pregnant women might be responsible for this reversal in the autonomic response to lying down. PMID- 10364856 TI - Effect of nitric oxide predilution on inhaled nitrogen dioxide concentrations. AB - We examined the possibility that predilution of a concentrated nitric oxide (NO) source with nitrogen, before contact with oxygen, can reduce the inspired nitrogen dioxide (NO2) concentration during administration of nitric oxide. A Manley Blease and a Siemens Servo 900 C ventilator delivered 10, 20, 40, 60 and 80 parts per million (ppm) NO using an NO source of 1000, 400 and 200 ppm. With the Manley Blease system, predilution from 1000 to 200 ppm NO reduced the inhaled NO2 concentration from 0.14 to 0.05 ppm (p < 0.01) at 10 ppm inhaled NO, and from 1.20 to 1.00 ppm (p < 0.01) at 40 ppm inhaled NO. With the Siemens Servo 900 C ventilator, inspiratory NO2 concentrations decreased from 0.21 to 0.11 ppm (p < 0.01) at 10 ppm inhaled NO, and from 1.49 to 1.16 ppm (p < 0.01) at 40 ppm NO. Predilution from 1000 to 400 ppm NO reduced the inspired NO2 concentrations by < 3% using either ventilator when the inspirated NO concentration was 80 ppm. Predilution of NO with nitrogen significantly reduced the inspired NO2 concentrations for nitric oxide concentrations between 10 and 40 ppm, but offered no clinically relevant advantage at higher NO concentrations. PMID- 10364857 TI - Cost-effectiveness of ondansetron for postoperative nausea and vomiting. AB - The decision as to whether prophylaxis against postoperative nausea and vomiting is better than treatment of established postoperative nausea and vomiting could be made on the basis of cost-effectiveness. The cost-effectiveness of ondansetron was calculated using data from published quantitative systematic reviews of randomised trials. Milligrams of ondansetron required to achieve a desired endpoint were chosen as a cost unit. Modelling was based on a cohort of 1000 patients, and examined control event rates (i.e. incidence of postoperative nausea and vomiting without prophylaxis) of between 10 and 90%. In a sensitivity analysis, cost-effectiveness of recommended intravenous doses (4 mg for treatment and prophylaxis) was compared with minimal effective doses as shown by meta analysis (1 mg for treatment, 8 mg for prophylaxis). Fewer patients experience any postoperative nausea and vomiting symptoms with prophylaxis compared with treatment. But prophylaxis is only marginally more effective than treatment, and treatment of established postoperative nausea and vomiting with effective doses (i.e. 1 or 4 mg) is more cost-effective and safer than prophylaxis with effective doses (i.e. 4 or 8 mg). Reasons for this are the selective treatment of patients who actually need treatment, the high success rate with a lowest dose tested (1 mg) in established postoperative nausea and vomiting, and the disappointing antinausea effect of prophylactic ondansetron even at an eight-fold higher dose. PMID- 10364858 TI - Peri-operative silent myocardial ischaemia in patients undergoing lower limb joint replacement surgery: an indicator of postoperative morbidity or mortality? AB - One hundred and twenty-seven patients undergoing major lower limb joint replacement surgery were studied to determine the incidence of silent myocardial ischemia and to ascertain any link between pre-operative cardiac risk factors, silent myocardial ischaemia and postoperative morbidity. Patients underwent ambulatory ECG monitoring for 4 days (on the pre-operative night and for 3 days postoperatively). Postoperative cardiorespiratory symptomatology and morbidity was assessed by questionnaire at 3 months. Eighty-seven patients had risk factors for silent myocardial ischaemia; 42 patients (30 with risk factors) had peri operative silent myocardial ischaemia. The median ischaemic loads (range) were 1.04 (0.32-13.31) min.h-1 pre-operatively and 5.53 (0.26-56.39), 6.69 (0.04 42.71) and 1.23 (0.1-53.74) min.h-1 on postoperative days 1-3, respectively. Risk factors did not predict the occurrence of silent myocardial ischaemia or an increased ischaemic load pre-operatively or overall postoperatively. New symptoms (chest pain, palpitations, breathlessness or fatigue) were associated with both silent myocardial ischaemia and ischaemic load (p < 0.05). Thus cardiac risk factors do not predict the occurrence of silent myocardial ischaemia or adverse outcome. Peri-operative silent myocardial ischaemia was associated with increased postoperative fatigue. PMID- 10364859 TI - The effect of intrathecal magnesium sulphate on nociception in rat acute pain models. AB - We examined the antinociceptive effect of intrathecally administered magnesium sulphate (MgSO4) in rats, using acute pain models including mechanical pressure, heat and subcutaneous formalin injection. According to the locomotion test 10 microliters of 6.2% MgSO4 did not produce motor paralysis. At the same dose, responses to pressure and heat were intact, compared with controls given saline. MgSO4 produced depression of pain responses only after the first 10 min in the formalin test. Our studies indicated that MgSO4 did not show remarkable antinociceptive effects in acute pain models. PMID- 10364860 TI - Comparison of sevoflurane and propofol with rocuronium for modified rapid sequence induction of anaesthesia. AB - We compared the use of sevoflurane and propofol with three different doses of rocuronium for modified rapid-sequence induction of anaesthesia. One hundred and forty adult patients were randomly allocated to have a rapid-sequence intravenous induction with propofol 2-3 mg.kg-1 (group P) or an inhalational induction with sevoflurane 8% in oxygen, using a vital capacity technique (group S). Following loss of the eyelash reflex, cricoid pressure was applied and 20 patients in each group were administered rocuronium 0.3 (groups P/0.3 and S/0.3), 0.45 (groups P/0.45 and S/0.45) or 0.6 (groups P/0.6 and S/0.6) mg.kg-1. An additional 10 patients in each group received only saline placebo in place of the muscle relaxant (groups P/Saline and S/Saline). Laryngoscopy was started 60 s later and intubating conditions evaluated by a blinded anaesthetist according to a standard scoring system. Intubating conditions were acceptable in one patient and no patient, respectively, following induction with sevoflurane and propofol without the muscle relaxant. The conditions were acceptable in 30, 55 and 90% of subjects with sevoflurane induction, and in 45, 80 and 90% of subjects with propofol induction following 0.3, 0.45 and 0.6 mg.kg-1 of rocuronium, respectively (no significant difference for each dose of rocuronium). The present study shows that intubating conditions during a rapid-sequence induction using rocuronium 0.6 mg.kg-1 following induction of anaesthesia with sevoflurane or propofol are similar. PMID- 10364861 TI - Continuous pulse oximetry in the general surgical ward: Nellcor N-200 versus Nellcor N-3000. AB - New measurement principles in pulse oximetry have been introduced to decrease the incidence of false movement alarms. Experimental studies have shown that the new Nellcor Symphony N-3000 may reduce the incidence of false alarms when monitoring during different activities. We compared the Nellcor Symphony N-3000 with the Nellcor N-200 pulse oximeter, when monitoring patients in the general surgical ward. Twenty-two patients were monitored during unrestricted ward activities for a total of 275 h with a N-3000 and a N-200 pulse oximeter simultaneously. Data were analysed for lack of concordance between the two pulse oximeters with respect to frequency of registered hypoxaemic episodes and thus the amount of time spent in the alarm state. The median number of desaturation episodes with the N-200 was 18 (range 0-511) compared with four (range 0-476) with the N-3000 (p < 0.0001). The median number of drop-outs (loss of signal) was 13 (range 1-46) with the N-200 compared with nine (2-41) with the N-3000 (p = 0.06). The N-200 registered saturation values of 85% or below for 23% of the observation time compared with 6% of the observation time with the N-3000 pulse oximeter (p < 0.0001). The different working principles of the two generations of oximeters were reflected in the present results derived from a clinical setting. The Nellcor Symphony N-3000 may offer an advantage compared with the Nellcor N-200, because of the reduced frequency of alarms and total time in alarm when monitoring patients in the general surgical ward. PMID- 10364862 TI - Prevention of needle-stick injury. Efficacy of a safeguarded intravenous cannula. AB - One possible method of reducing the incidence of needle-stick injury is to use needles with safeguard mechanisms. The needle of the Insyte AutoGuard intravenous cannula can be retracted into the safety barrel. One hundred patients were randomly allocated to receive either an 18-gauge conventional Insyte intravenous cannula (group C) or the AutoGuard cannula (group AG) to assess the ease of use and efficacy of the AutoGuard device. It was possible to insert the cannula into the vein within two attempts in all patients; there was no significant difference between two groups with respect to ease of insertion. No problems, such as inadvertent withdrawal of the needle, occurred during insertion in any patient. Handling the withdrawn needle was judged significantly safer in group AG than in group C (p < 0.001). Blood contamination often occurred where a withdrawn needle was placed in group C, whereas no blood stain was detected in any case in group AG (p < 0.001). The AutoGuard cannula provides safer handling of a withdrawn needle without reducing its ease of insertion. PMID- 10364863 TI - A bronchogenic cyst in an infant causing tracheal occlusion and cardiac arrest. AB - A 3-month-old infant treated for 3 weeks for suspected bronchiolitis, developed episodes of profound desaturation. A lateral X-ray showed displacement and compression of the trachea. Respiratory arrest, from which she was successfully resuscitated, occurred just before MRI scan. The mass was removed at thoracotomy and a histological diagnosis of a bronchogenic cyst was made. Mediastinal masses in babies are relatively rare, and the situation in which they present with acute respiratory distress may prove extremely challenging to the anaesthetist. Bronchogenic cysts are difficult to diagnose pre-operatively and awareness may assist in the peri-operative management of these infants. PMID- 10364864 TI - Prophylactic anti-emetic efficacy of ondansetron in laparoscopic cholecystectomy under total intravenous anaesthesia. A randomised, double-blind comparison with droperidol, metoclopramide and placebo. AB - The prophylactic anti-emetic efficacy and safety of pre-operative intravenous ondansetron was evaluated in a randomised, double-blind, comparison with droperidol, metoclopramide and placebo in 160 ASA grade 1 and 2 patients undergoing laparoscopic cholecystectomy under total intravenous anaesthesia. The patients were randomly allocated to receive ondansetron (4 mg), droperidol (1.25 mg), metoclopramide (10 mg) or placebo given as a single intravenous dose immediately before induction of a standardised general anaesthetic. There were no significant differences between the four study groups with regard to the demographic and anaesthetic data, postoperative analgesia, postoperative sedation scores, duration of postoperative hospital stay and incidence of adverse events. The incidence of nausea and vomiting was significantly lower (p < 0.05) between 1 h and 4 h after surgery in the ondansetron group compared with the droperidol, metoclopramide and placebo groups. The incidence of nausea was similar in the four groups in the other study periods: 0-1 h and 4-24 h. The incidence of vomiting was lower in the ondansetron, droperidol and metoclopramide groups than in the placebo group between 1 and 4 h but was the same between 4 and 24 h. As a result of the lower incidence of nausea and vomiting between 1 h and 4 h in the ondansetron group, the overall incidence of nausea and vomiting was lower during the first 24 h after surgery in this group than in the other three groups. PMID- 10364865 TI - Conditions for laryngeal mask insertion. A comparison of propofol versus sevoflurane with or without alfentanil. AB - One hundred unpremedicated ASA 1 or 2 patients scheduled for elective surgery were divided equally into four groups and recruited into this prospective, randomised parallel groups study. Induction was with propofol 2.5 mg.kg-1 or vital-capacity breath induction with sevoflurane (> 7% in the inspiratory gas) in 65% nitrous oxide and oxygen, or gaseous induction with sevoflurane plus alfentanil 5 micrograms.kg-1 or propofol 2.5 mg.kg-1 and alfentanil 5 micrograms.kg-1. The conditions for laryngeal mask insertion were assessed and graded on a three-point scale using six variables. The overall condition for laryngeal mask insertion was assessed as excellent, satisfactory or poor on the basis of total score in each group. Excellent or satisfactory conditions were observed in 25 (100%) patients in the sevoflurane-alfentanil group, 22 (88%) in the propofol-alfentanil group and 16 (64%) patients each in the propofol and sevoflurane groups (p < 0.001). A sevoflurane-alfentanil combination provides better conditions for laryngeal mask insertion when compared with sevoflurane alone, or a propofol-alfentanil combination. PMID- 10364866 TI - Influence of induction of anaesthesia on intubating conditions one minute after rocuronium administration: comparison of ketamine and thiopentone. AB - We compared the effect of thiopentone and ketamine on intubating conditions after rocuronium 0.6 mg.kg-1 in two groups of patients (n = 16 each), aged 21-44 years, undergoing elective surgery. Premedication consisted of alprazolam 1 mg by mouth 1 h before surgery. All patients received midazolam 2 mg intravenously 2 min before intravenous administration of thiopentone 5 mg.kg-1 or ketamine 2.5 mg.kg 1. Muscle relaxation was provided by rocuronium 0.6 mg.kg-1. One minute after rocuronium administration, tracheal intubation was performed within 15 s by a skilled anaesthetist blinded to the treatment group assignment. Intubating conditions were graded as excellent, good, fair or poor on the basis of jaw relaxation, position of vocal cords and diaphragmatic response. Neuromuscular transmission was assessed at the adductor pollicis muscle using a TOF-GUARD monitor. Excellent and good intubating conditions were obtained in 100% of patients in the ketamine group and in 50% of patients in the thiopentone group (p = 0.002). Jaw relaxation was similar in both groups but vocal cord conditions were better and the diaphragmatic response less marked in the ketamine group compared with the thiopentone group (p = 0.002). The degree of neuromuscular block [% decrease of T1, mean (SD)] at the time of intubation was similar: 51.8 (25)% (ketamine group) and 54.3 (23.1)% (thiopentone group). We conclude that ketamine 2.5 mg.kg-1 provides better intubating conditions than thiopentone 5 mg.kg-1 1 min after administration of rocuronium 0.6 mg.kg-1. PMID- 10364867 TI - The immediate impact of opening an adult high dependency unit on intensive care unit occupancy. AB - We assessed the hourly occupancy of our intensive care and high dependency units over an 8-week period commencing on the day our high dependency unit opened. Using criteria established by the working group on 'Guidelines on Admission to and Discharge from Intensive Care and High Dependency Units' published by the National Health Service Executive, we defined each patient daily as intensive care or high dependency status. Compared with hourly occupancy figures obtained before the high dependency unit opened, occupancy of the intensive care unit by high dependency patients has been shown to decrease significantly from 21.6% to 11.2%. Use of intensive care beds became more appropriate, their occupancy increasing significantly from 63.7% to 73.4%. A significant decrease in readmissions occurred, supporting the hypothesis that having high dependency beds reduces the number of patients discharged prematurely to the wards. PMID- 10364868 TI - Meeting the standards for interhospital transfer of adults with severe head injury in the United Kingdom. AB - In December 1996, the Association of Anaesthetists of Great Britain and Ireland produced a series of recommendations outlining the safe conduct of interhospital transfers for patients with acute head injuries. We assessed the current ability of UK hospitals to implement these recommendations and opinions on the formation of transfer teams, using a postal questionnaire. This was sent to all Royal College of Anaesthetists tutors, 268 of whom replied (94% response rate). Of the hospitals surveyed, 208 received adult head-injury patients but did not have on site neurosurgical facilities. In 171 (86.8%) of these hospitals, senior house officers could be expected to accompany the patient during subsequent transfer. The majority of hospitals (192, 92.3%) were able to monitor ECG, pulse oximetry and blood pressure during the journey, but only 97 (46.6%) had facilities to monitor end tidal carbon dioxide levels. As a result of the anaesthetist's involvement in the transfer, emergency operating could be delayed in 169 (81.3%) hospitals. One hundred and fifty-eight (76%) respondents thought that the formation of transfer teams to transport critically ill patients would have some merit. Hospitals are responding to the published guidelines, but improvements are still needed in levels of equipment and insurance provision, along with the identification of a designated consultant at each hospital with responsibility for transfers. PMID- 10364869 TI - The use of tramadol following day--case oral surgery. AB - This prospective, randomised double-blinded study was designed to assess the analgesic efficacy and occurrence of nausea when tramadol is added to a nonsteroidal anti-inflammatory drug to provide analgesia following day-case third molar teeth extraction. All patients received oral diclofenac pre-operatively and one of four treatments intra-operatively: fentanyl and metoclopramide, tramadol and metoclopramide, fentanyl and ondansetron, or tramadol and ondansetron. There were no significant differences between groups in scores for pain in the early postoperative period. However, there were significant differences in nausea scores at this time, with the fentanyl-ondansetron group having the lowest and the tramadol-ondansetron group having the highest scores. There were no significant differences in the incidence of pain or nausea in the following 24 h. We conclude that the addition of tramadol to diclofenac results in no useful improvement in analgesic effect, and that the use of ondansetron fails to reduce the nausea associated with tramadol. PMID- 10364870 TI - Attitudes to oral feeding following caesarean section. AB - In order to evaluate postoperative nutrition in women who have undergone Caesarean section, we conducted a national survey. Questionnaires were sent to 100 randomly selected obstetric units in the UK, and were completed and returned by senior midwives. We found that that only 21.5% of units had a departmental policy concerning feeding after Caesarean section. Midwives decided when women could eat and drink in the majority of obstetric units (78.5%), often without the help of guidelines. The period of postoperative starvation was found to vary greatly, from < 1 h in some units to > 24 h in others. We suggest that all obstetric units should produce guidelines in order to rationalise postoperative feeding for women following Caesarean section. PMID- 10364871 TI - Central neural blockade and the compartment syndrome. PMID- 10364872 TI - Caudal analgesia in children. PMID- 10364873 TI - Oral versus rectal diclofenac for postoperative tonsillectomy pain in children. PMID- 10364874 TI - Acute pain service audit. PMID- 10364875 TI - Combined spinal epidural anaesthesia. PMID- 10364876 TI - Combined spinal epidural anaesthesia. PMID- 10364877 TI - Epidural blood patch. PMID- 10364878 TI - Sevoflurane induction and acute epiglottitis. PMID- 10364879 TI - Global pollution--the anaesthetist's contribution. PMID- 10364880 TI - Sudden death from inhalation of petrol vapour. PMID- 10364881 TI - Magnesium sulphate for the control of spasms in severe tetanus. PMID- 10364882 TI - Gastro-esophageal reflux during day case gynaecological laparoscopy. PMID- 10364883 TI - The Combitube and cervical spine immobilisation. PMID- 10364884 TI - A close shave. PMID- 10364885 TI - Grading of direct laryngoscopy. PMID- 10364886 TI - The intubating laryngeal mask (ILMA) in an emergency failed intubation. PMID- 10364887 TI - Awareness during cardiac anaesthesia. PMID- 10364888 TI - Raynaud's phenomenon and propofol. PMID- 10364889 TI - An absolute contraindication to nitrous oxide. PMID- 10364891 TI - Humouring the patient. PMID- 10364890 TI - Another cracking idea. PMID- 10364892 TI - A slap on the back. PMID- 10364894 TI - A gastronomic airway assessment. PMID- 10364893 TI - Prolonged pharyngeal obstruction after the Heimlich manoeuvre. PMID- 10364895 TI - Music in theatre. PMID- 10364896 TI - Fatal myocardial necrosis. PMID- 10364897 TI - Prevalence of hip osteoarthritis in Iceland. AB - OBJECTIVE: To assess the prevalence of primary hip osteoarthritis (OA) in Iceland. To compare the prevalence of primary hip OA in Iceland with published rates of primary hip OA for related Scandinavian populations. METHODS: Roentgenographs were examined of 1530 Icelandic people 35 years or older (653 males, 877 females) subjected to colon radiography during the years 1990-1996. The radiographs examined represent approximately 40% of all colon radiographs taken in Iceland during this period. After exclusion of non-primary hip OA cases, the minimum hip joint space was measured with a mm ruler. Presence of hip OA was defined as a minimum joint space of 2.5 mm or less on an anteroposterior radiograph. Intraclass correlation coefficients for inter and intraobserver variability of assessment of mm joint space were 0.91 and 0.95, respectively. RESULTS: Of the 1517 people included, 227 hips in 165 patients (77 men, 88 women) were diagnosed as having radiological primary hip OA. The mean age at colon examination for these patients was 68 (35-89) years. The overall prevalence of coxarthrosis among all examined patients 35 years and older was 10.8% (12% for men, 10% for women), rising from 2% at 35-39 years to 35.4% for those 85 years or older. If the population structure (age and sex distribution) for those older than 35 years in Iceland was used to standardise prevalence for both Iceland and south Sweden (using previously published data for south Sweden), the age and sex standardised prevalence of hip OA for those older than 35 years in Iceland was 8%, compared with 1.2% for south Sweden. CONCLUSIONS: The prevalence of radiological primary hip OA is very high in Iceland, and in excess of fivefold higher than the prevalence found by using similar techniques in studies on related populations in southern Scandinavia. The rate difference is particularly notable for those younger than 70 years. PMID- 10364899 TI - Clinical features and outcome of septic arthritis in a single UK Health District 1982-1991. AB - AIMS: To determine the clinical features of a large number of unselected UK hospital patients with confirmed septic arthritis and to determine those features associated with a poor outcome. STUDY DESIGN: Retrospective, case-note survey. SETTING: A single English Health District. PATIENTS: All patients admitted to hospital in Nottingham during the period 1 January 1982 to 31 December 1991 with confirmed septic arthritis were included. OUTCOME MEASURES: Death, osteomyelitis and recorded functional impairment. RESULTS: The spectrum of causative organisms remains similar to that seen in previous studies with the Gram positive organisms Staphylococcus aureus and Streptococci responsible for 74% of cases, gonococcal infections though were less common. Culture of joint aspirates and or blood were positive in 82% of cases, with the Gram stain demonstrating the causative organism in 50% of cases. Pre-existing joint disease was evident in 35% of cases. The mortality remains high at 11.5% with a significant additional morbidity of 31.6%. Multivariate analysis suggests that important predictors of death are: confusion at presentation, age > or = 65 years, multiple joint sepsis or involvement of the elbow joint, and of morbidity are: age > or = 65 years, diabetes mellitus, open surgical drainage, and Gram positive infections other than S aureus. CONCLUSIONS: Septic arthritis continues to be associated with a considerable degree of morbidity and mortality. These results confirm the importance of obtaining synovial fluid and blood for culture before starting antimicrobial treatment. The apparent poorer outcome found with surgical intervention is in line with some previous suggestions but should be interpreted with caution in light of the retrospective nature of this study. PMID- 10364898 TI - Validity of histopathological grading of articular cartilage from osteoarthritic knee joints. AB - OBJECTIVES: To determine the validity of the histological-histochemical grading system (HHGS) for osteoarthritic (OA) articular cartilage. METHODS: Human articular cartilage was obtained from macroscopically normal (n = 13) and OA (n = 21) knee joints. Sections of central and peripheral regions of normal samples were produced. Sections of regions containing severe, moderate, and mild OA changes were produced from each OA sample. A total of 89 sections were graded by means of the HHGS (0-14) twice by three observers. RESULTS: Average scores for regions designated severe (8.64) and moderate (5.83) OA were less than the expected (10-14 and 6-9, respectively) according to the HHGS, whereas average scores for the region designated mild (5.29) OA and central and peripheral regions (2.19) of normal cartilage were higher than expected (2-5 and 0-1, respectively). The HHGS was capable of differentiating between articular cartilage from macroscopically normal and OA joints and between the region designated severe OA and other regions. However, the HHGS did not adequately differentiate between regions designated mild and moderate OA. Values for sensitivity, specificity, and efficiency for all regions varied considerably. CONCLUSION: The HHGS is valid for normal and severe OA cartilage, but does not permit distinction between mild and moderate OA changes in articular cartilage. PMID- 10364900 TI - Combination therapy in early rheumatoid arthritis: a randomised, controlled, double blind 52 week clinical trial of sulphasalazine and methotrexate compared with the single components. AB - OBJECTIVES: To investigate the potential clinical benefit of a combination therapy. METHODS: 205 patients fulfilling the ACR criteria for rheumatoid arthritis (RA), not treated with disease modifying antirheumatoid drugs previously, with an early (< or = 1 year duration), active (Disease Activity Score (DAS) > 3.0), rheumatoid factor and/or HLA DR 1/4 positive disease were randomised between sulphasalazine (SASP) 2000 (maximum 3000) mg daily (n = 68), or methotrexate (MTX) 7.5 (maximum 15) mg weekly (n = 69) or the combination (SASP + MTX) of both (n = 68). RESULTS: The mean changes in the DAS during the one year follow up of the study was -1.15, -0.87, -1.26 in the SASP, MTX, and SASP + MTX group respectively (p = 0.019). However, there was no statistically significant difference in terms of either EULAR good responders 34%, 38%, 38% or ACR criteria responders 59%, 59%, 65% in the SASP, MTX, and SASP + MTX group respectively. Radiological progression evaluated by the modified Sharp score was very modest in the three groups: mean changes in erosion score: +2.4, +2.4, +1.9, in narrowing score: +2.3, +2.1, +1.6 and in total damage score: +4.6, +4.5, +3.5, in the SASP, MTX, and SASP + MTX groups respectively. Adverse events occurred more frequently in the SASP + MTX group 91% versus 75% in the SASP and MTX group (p = 0.025). Nausea was the most frequent side effect: 32%, 23%, 49% in the SASP, MTX, and SASP + MTX groups respectively (p = 0.007). CONCLUSION: This study suggests that an early initiation therapy of disease modifying drug seems to be of benefit. However, this study was unable to demonstrate a clinically relevant superiority of the combination therapy although several outcomes were in favour of this observation. The tolerability of the three treatment modalities seems acceptable. PMID- 10364901 TI - Clinical and laboratory manifestations of elderly onset psoriatic arthritis: a comparison with younger onset disease. AB - OBJECTIVE: Although the influence of age on clinical and laboratory features has been widely demonstrated in many arthropathies, studies on elderly onset (> 60 years) psoriatic arthritis (EOPsA) are rare. This study compares manifestations at onset and two year outcome of EOPsA with those of younger onset PsA (YOPsA). PATIENTS AND METHODS: Sixty-six consecutive PsA patients with disease duration < 1 year, 16 EOPsA (> 60 years) and 50 YOPsA (< or = 60 years) were admitted to a prospective study. Clinical, laboratory, and radiographic assessment were carried out at admission and after two years. HLA class I and bone scintigraphy were also recorded. In 10 patients with EOPsA and 24 with YOPsA it was possible to obtain synovial fluid, which was subsequently analysed for local inflammatory indices, including interleukin (IL) 1 beta, IL6, and IL8. RESULTS: Presenting manifestations of EOPsA differed from YOPsA in number of active joints (mean (SD)) (12.2 (6.3) v 6.7 (4.6), p < 0.001), foot bone erosions (2.7 (1.2) v 1.1 (1.1), p < 0.001), erythrocyte sedimentation rate (64.2 (35.3) v 30.5 (30.0) mm 1st h, p < 0.001), C reactive protein (3.9 (2.0) v 1.3 (1.3) mg/dl, p < 0.001) and synovial fluid IL1 beta (8.0 (4.7) v 3.0 (3.0) pg/ml, p < 0.001) and IL6 (828.2 (492.6) v 469.3 (201.4) pg/ml, p < 0.005). No differences were found in the number of subjects with dactylitis, pitting oedema, HLA-B27, or signs of sacroiliac and sternoclavicular joint involvement at bone scintigraphy. After two years, progression was more evident in EOPsA than in YOPsA, as the number of new erosions in the hands and also the C reactive protein were higher in EOPsA patients. CONCLUSION: PsA has a more severe onset and a more destructive outcome in elderly people (onset > 60 years) than in younger subjects. This behaviour may be influenced by immune changes associated with aging, as suggested by the higher concentrations of IL1 beta and IL6 found in the synovial fluid of EOPsA than in YOPsA. PMID- 10364904 TI - Are radiological joint space widths of normal hips asymmetrical? AB - BACKGROUND: To be certain that the joint space width is abnormal in the case of hip joint pain when compared with the contralateral hip requires knowledge of physiological dissymmetry. AIM OF THE STUDY: To assess interindividual variability and dissymmetry in pelvic radiological joint space width. METHODS: Pelvic radiographs of subjects without hip joint disease. Measurement with a 0.1 mm graduated magnifying glass and 0.5 mm graduated flat ruler at the hip superointermediate site (vertical going through the femoral head centre). After randomisation of the side to measure, analysis of nine groups of 19 plain films by one investigator blind for the result of the contralateral side. RESULTS: The difference between the left and right hip was plotted against the corresponding mean for all 171 normal subjects. This shows the frequency and the limits of the asymmetry at each measurement site. The asymmetry is independent of interindividual variability of the joint space width and greater than the measurement error in most subjects. CONCLUSION: This study confirms the interindividual variability of hip joint space width, shows the frequency of hip joint space asymmetry and defines its limit. PMID- 10364902 TI - Remitting seronegative symmetrical synovitis with pitting oedema (RS3PE) syndrome: a prospective follow up and magnetic resonance imaging study. AB - OBJECTIVE: To determine the clinical characteristics of patients with "pure" remitting seronegative symmetrical synovitis with pitting oedema (RS3PE) syndrome, and to investigate its relation with polymyalgia rheumatica (PMR). Magnetic resonance imaging (MRI) was used to describe the anatomical structures affected by inflammation in pure RS3PE syndrome. METHODS: A prospective follow up study of 23 consecutive patients with pure RS3PE syndrome and 177 consecutive patients with PMR diagnosed over a five year period in two Italian secondary referral centres of rheumatology. Hands or feet MRI, or both, was performed at diagnosis in 7 of 23 patients. RESULTS: At inspection evidence of hand and/or foot tenosynovitis was present in all the 23 patients with pure RS3PE syndrome. Twenty one (12%) patients with PMR associated distal extremity swelling with pitting oedema. No significant differences in the sex, age at onset of disease, acute phase reactant values at diagnosis, frequency of peripheral synovitis and carpal tunnel syndrome and frequency of HLA-B7 antigen were present between patients with pure RS3PE and PMR. In both conditions no patient under 50 was observed, the disease frequency increased significantly with age and the highest frequency was present in the age group 70-79 years. Clinical symptoms for both conditions responded promptly to corticosteroids and no patient developed rheumatoid arthritis during the follow up. However, the patients with pure RS3PE syndrome were characterised by shorter duration of treatment, lower cumulative corticosteroid dose and lower frequency of systemic signs/symptoms and relapse/recurrence. Hands and feet MRI showed evidence of tenosynovitis in five patients and joint synovitis in three patients. CONCLUSION: The similarities of demographic, clinical, and MRI findings between RS3PE syndrome and PMR and the concurrence of the two syndromes suggest that these conditions may be part of the same disease and that the diagnostic labels of PMR and RS3PE syndrome may not indicate a real difference. The presence of distal oedema seems to indicate a better prognosis. PMID- 10364905 TI - Assessment of mutilans-like hand deformities in chronic inflammatory joint diseases. A radiographic study of 52 patients. AB - OBJECTIVES: To evaluate patients with mutilans-like hand deformities in chronic inflammatory joint diseases and to determine radiographic scoring systems for arthritis mutilans (AM). METHODS: A total of 52 patients with severe hand deformities were collected during 1997. A Larsen hand score of 0-110 was formed to describe destruction of the hand joints. Secondly, each ray of the hand was assessed individually by summing the Larsen grade of the wrist and the grades of the MCP and PIP joints. When the sum of these grades was > or = 13, the finger was considered to be mutilated. A mutilans hand score of 0-10 was formed according to the number of mutilans fingers. Surgical treatment and spontaneous fusions were recorded. RESULTS: The study consisted of 22 patients with juvenile rheumatoid arthritis (JRA), nine with rheumatoid factor (RF) positive and 13 with RF negative arthritis, 27 patients with RF positive RA, and three adult patients with other diagnoses. The mean age of patients with adult rheumatic diseases was 27 years at the onset of arthritis. The mean disease duration in all patients was 30 years. The mean Larsen hand score was 93. Four patients had no mutilans fingers and in 15 patients all 10 fingers were mutilated. The Larsen hand score of 0-110 and the mutilans hand score of 0-10 correlated well (rs = 0.90). Fourteen patients showed spontaneous fusions in the peripheral joints. A total of 457 operations were performed on 48 patients. CONCLUSION: Both the Larsen hand score of 0-110 and the mutilans hand score of 0-10 improve accuracy in evaluating mutilans-like hand deformities, but in unevenly distributed hand deformities the mutilans hand score is better in describing deformation of individual fingers. PMID- 10364903 TI - Monocyte activation in patients with Wegener's granulomatosis. AB - OBJECTIVE: Wegener's granulomatosis (WG) is an inflammatory disorder characterised by granulomatous inflammation, vasculitis, and necrotising vasculitis and is strongly associated with anti-neutrophil cytoplasmic antibodies (ANCA). Activated monocytes/macrophages are present in renal biopsy specimens and participate in granuloma formation by synthesising and secreting a variety of chemoattractants, growth factors, and cytokines. In view of these findings, in vivo monocyte activation was evaluated in patients with WG and the findings related to parameters of clinical disease activity. METHODS: Monocyte activation was analysed by measuring plasma concentrations of soluble products of monocyte activation, that is neopterin and interleukin 6 (IL6), by ELISA, and by quantitating the surface expression of activation markers on circulating monocytes by flow cytometry. RESULTS: Twenty-four patients with active WG were included in this study. Ten of these patients were also analysed at the time of remission. Twelve patients with sepsis served as positive controls, and 10 healthy volunteers as negative controls for monocyte activation. Patients with active disease had increased monocyte activation compared with healthy controls as shown by increased concentrations of neopterin (p < 0.0001) and increased surface expression of CD11b (p < 0.05) and CD64 (p < 0.05). In those patients with increased concentrations of IL6 during active disease plasma concentrations of IL6 decreased during follow up when patients went into remission (p < 0.0001). In addition, neopterin (r = 0.37, r = 0.44), IL6 (r = 0.37, r = 0.60) and CD63 expression (r = 0.39, r = 0.45) correlated significantly with disease activity as measured by the Birmingham Vasculitis Activity Score and C reactive protein values, respectively. Compared with patients with sepsis, all markers of monocyte activation in patients with vasculitis were lower. CONCLUSION: It is concluded that disease activity in WG correlates with the extent of activation of monocytes, compatible with their role in the pathophysiology of this disease. PMID- 10364906 TI - Hydroxychloroquine treatment for primary Sjogren's syndrome: its effect on salivary and serum inflammatory markers. AB - OBJECTIVE: To evaluate the effect of hydroxychloroquine treatment on interleukin 6 (IL6), hyaluronic acid (HA), and soluble interleukin 2 receptor (sIL2R) concentrations in the saliva and serum of patients with primary Sjogren's syndrome (SS). METHODS: Fourteen SS patients treated with hydroxychloroquine 200 mg/day for 12 months were investigated in an open prospective study. Clinical parameters of efficacy and routine biochemical and haematological data to assess drug safety and tolerability were determined every three months. Salivary and serum IL6, sIL2R, and HA values were determined at study entry, 6 and 12 months, using ELISA and radiometric assays. RESULTS: After hydroxychloroquine treatment, salivary IL6 concentrations decreased from 13.2 (1.2) to 7.3 (1.1) pg/ml (mean (SEM)) (p < 0.0001). Similarly, salivary HA concentrations were also reduced from 577.8 (120) to 200 (34) ng/ml (mean (SEM) (p < 0.003). Serum IL6 concentrations decreased from 5.4 (0.6) to 2.9 (0.2) pg/ml (mean (SEM) (p < 0.001), while serum HA concentrations remained unchanged. No change has been detected in salivary or serum sIL2R concentrations after 12 months of treatment with hydroxychloroquine. Treatment also resulted in significant reduction in erythrocyte sedimentation rate, serum gamma globulin, and C reactive protein values while only partial clinical improvement was noted in some patients. A more pronounced decrease of salivary IL6 and HA levels was found in the two patients in whom a reduction in the swelling of the parotid gland was noted. CONCLUSION: In this open label study of hydroxychloroquine treatment for SS a significant reduction of some salivary inflammatory markers was seen at the end of 12 months. Although during the treatment period only a partial clinical effect could be noted, the findings suggest that a double blind controlled study of hydroxychloroquine in SS is indicated. PMID- 10364907 TI - Effects of nabumetone compared with naproxen on platelet aggregation in patients with rheumatoid arthritis. AB - OBJECTIVE: To test the hypothesis that nabumetone (a partially selective cyclooxygenase-(COX)-2 inhibitor) has less effect on platelet aggregation than naproxen (a non-selective COX-inhibitor) in patients with rheumatoid arthritis (RA). METHODS: A crossover study in 10 RA patients was performed, using either nabumetone or naproxen for two weeks, and, after a washout period of two weeks, the other drug during another two weeks. Platelet aggregation studies were performed and bleeding time was assessed before and after each treatment period. RESULTS: Maximum platelet aggregation induced by epinephrine and by collagen was significantly more reduced after the use of naproxen than of nabumetone; secondary aggregation induced by ADP and epinephrine disappeared more often by naproxen than by nabumetone. Bleeding times were not influenced. CONCLUSION: COX dependent platelet aggregation in RA patients seems to be more inhibited by naproxen than by nabumetone. This may be relevant for patients requiring non steroidal anti-inflammatory drug treatment but who have an increased risk of bleeding as well. PMID- 10364908 TI - Guidelines for rheumatology undergraduate core curriculum. EULAR Standing Committee on Education and Training. PMID- 10364909 TI - Immunological evaluation of cytokine and anticytokine immunotherapy in vivo: what have we learnt? PMID- 10364910 TI - Polyarthritis and pitting oedema. PMID- 10364911 TI - Hip pain. PMID- 10364912 TI - Osteoarthrosis of the knee in men and women in association with overweight, smoking, and hormone therapy. AB - OBJECTIVES: The aim was to examine the relation between osteoarthrosis of the knee leading to prosthetic surgery among men and women and overweight, smoking, and hormone therapy. METHODS: A case-referent study was performed with a study base of all men and women, born 1921-1938, living in 14 counties in Sweden during 1991-95. The cases (n = 625) were identified through the Swedish Knee Arthroplasty Register. The referents (n = 548) were randomly selected through the central population register from the same counties. Detailed information on general health status, height, weight, smoking habits, medication, use of hormones, specific physical loads from occupation and housework, and sports activities was collected by a telephone interview and a postal questionnaire. The cases were classified in terms of high, medium or low/non-exposure to the factors studied, according to the distribution of variables among the referents. RESULTS: Women with high body mass index (BMI) at the age of 40 had a relative risk of 9.2 (95% CI 5.3, 16.0) of developing severe knee osteoarthrosis later in life, and for men at the same age the relative risk was 3.9 (95% CI 2.3, 6.4). Smokers were less likely to develop severe knee osteoarthrosis compared with nonsmokers. Oestrogen therapy for women over 50 showed an increased relative risk of 1.8 (95% CI 1.2, 2.6), while use of oral contraceptives did not influence the risk. CONCLUSION: Overweight is a risk factor for knee osteoarthrosis leading to prosthetic surgery in men and women, with the strongest relation for women. Oestrogen therapy after 50 increased the relative risk, while smoking decreased it. PMID- 10364913 TI - Magnetic resonance imaging of the wrist in early rheumatoid arthritis reveals progression of erosions despite clinical improvement. AB - OBJECTIVES: To investigate the progression of joint damage in early rheumatoid arthritis (RA) using magnetic resonance imaging (MRI) of the wrist and determine whether this technique can be used to predict prognosis. METHODS: An inception cohort of 42 early patients has been followed up prospectively for one year. Gadolinium enhanced MRI scans of the dominant wrist were obtained at baseline and one year and scored for synovitis, tendonitis, bone marrow oedema, and erosions. Plain radiographs were performed concurrently and scored for erosions. Patients were assessed clinically for disease activity and HLA-DRB1 genotyping was performed. RESULTS: At one year, MRI erosions were found in 74% of patients (31 of 42) compared with 45% at baseline. Twelve patients (28.6%) had radiographic erosions at one year. The total MRI score and MRI erosion score increased significantly from baseline to one year despite falls in clinical measures of inflammation including erythrocyte sedimentation rate (ESR), C reactive protein (CRP), and swollen joint count (p < 0.01 for all). Baseline findings that predicted carpal MRI erosions at one year included a total MRI score of 6 or greater (sensitivity: 93.3%, specificity 81.8%, positive predictive value 93.3%, p = 0.000007), MRI bone oedema (OR = 6.47, p < 0.001), MRI synovitis (OR = 2.14, p = 0.003), and pain score (p = 0.01). Radiological erosions at one year were predicted by a total MRI score at baseline of greater than 13 (OR = 12.4, p = 0.002), the presence of MRI erosions (OR = 11.6, p = 0.005), and the ESR (p = 0.02). If MRI erosions were absent at baseline and the total MRI score was low, radiological erosions were highly unlikely to develop by one year (negative predictive value 0.91 and 0.92 respectively). No association was found between the shared epitope and erosions on MRI (p = 0.4) or radiography (p = 1.0) at one year. CONCLUSIONS: MRI scans of the dominant wrist are useful in predicting MRI and radiological erosions in early RA and may indicate the patients that should be managed aggressively. Discordance has been demonstrated between clinical improvement and progression of MRI erosion scores. PMID- 10364914 TI - Association of polymorphism in glutathione S-transferase loci with susceptibility and outcome in rheumatoid arthritis: comparison with the shared epitope. AB - OBJECTIVE: To determine whether glutathione S-transferase GSTM1, GSTM3, GSTT1, and GSTP1 genotypes influence susceptibility or outcome in rheumatoid arthritis (RA). METHODS: 277 RA patients were compared with 577 controls to examine any associations between GST genotypes and susceptibility to RA. The effect of genotypes on outcome (Larsen and functional scores) and time integrated acute phase responses (erythrocyte sedimentation rate and C reactive protein) was assessed in 122 patients with disease duration of 5-10 years. GST and HLA-DRB1 genotypes were determined using polymerase chain reaction based assays. Data were analysed using multiple regression analysis with correction for age, sex, disease duration, and the DRB1 associated shared epitope (SE) and rheumatoid factor (RF) positivity where appropriate. RESULTS: The GSTM1*A/*B genotype was less common in RA cases (3 of 276) than in controls (22 of 591) (exact p = 0.047), though significance was lost when adjustment was made for multiple comparisons. The Larsen score was higher (p = 0.039) in the GSTM1 null patients (89.9) than those with other GSTM1 genotypes (74.7), and this was independent of the SE. Again, correction for multiple testing resulted in loss of significance. The difference in Larsen scores between patients homozygous or negative for the SE (87.9 v 74.3) was similar to that between GSTM1 null and non-null patients. No associations between GSTM3 or GSTT1 genotypes and disease markers were identified although the association between GSTP1*B/*B and Larsen score approached significance (p = 0.096). CONCLUSION: It is proposed that certain GSTs may influence susceptibility and radiological progression in RA and that this is independent of the effect of the HLA-DRB1 associated SE. The mechanism for this effect is presumed to be because of differences in the ability of various GST enzymes to utilise the cytotoxic products of oxidant stress. Although significance was lost after correction for multiple testing, the data indicate that further studies may be of value in RA to determine the influence of the GST and other genes involved in cellular protection against oxidative stress. PMID- 10364915 TI - Serum samples of patients with rheumatoid arthritis contain a specific autoantibody to "denatured" aldolase A in the osteoblast-like cell line, MG-63. AB - OBJECTIVE: To identify rheumatoid arthritis (RA) specific autoantibody and its antigen in the human osteoblast-like cell line, MG-63. METHODS: MG-63 cell extract was subjected to western blotting by using RA and normal serum samples as probes. The autoantigen was purified and its N-terminal sequence was determined by automated Edman degradation. The reactivity of denatured aldolase A was evaluated by immunoblotting. Screening by enzyme linked immunosorbent assay (ELISA) using the autoantibody was performed. RESULTS: 40 kDa protein was found only in the RA serum samples and it was identified as aldolase A. A polyclonal antibody for rabbit muscle aldolase A bound to the 40 kDa protein and reacted in preference with the denatured enzyme. Using ELISA for denatured rabbit aldolase A, the autoantibody was found in approximately 10% of RA patients, whereas it was not found in the other arthropathy and healthy adults. CONCLUSION: This 40 kDa anti-aldolase A autoantibody, which was identified only in serum samples of RA patients with severe bone erosion, could be related to a certain event that induces RA specific joint destructions. PMID- 10364916 TI - Complement C1s activation in degenerating articular cartilage of rheumatoid arthritis patients: immunohistochemical studies with an active form specific antibody. AB - OBJECTIVE: The first complement component C1s was reported to have novel functions to degrade matrix components, besides its activities in the classic complement pathway. This study explores participation of C1s in articular cartilage degradation in rheumatoid arthritis (RA). METHODS: Normal articular cartilage (n = 6) and cartilage obtained from joints with RA (n = 15) and osteoarthritis (OA, n = 10) were immunostained using anti-C1s monoclonal antibodies PG11, which recognises both active and inactive C1s, and M241, which is specifically reactive to activated C1s. The effects of inflammatory cytokines on C1s production by human articular chondrocytes were also examined by sandwich ELISA. RESULTS: In normal articular cartilage, C1s was negative in staining with both PG11 and M241. In contrast, degenerating cartilage of RA was stained with PG11 (14 of 15 cases), and in most of the cases (13 of 15 cases) C1s was activated as revealed by M241 staining. In OA, C1s staining was restricted in severely degrading part of cartilage (5 of 10 cases), and even in that part C1s was not activated. In addition, C1s production by chondrocytes in vitro was increased by an inflammatory cytokine, tumour necrosis factor alpha. CONCLUSION: These results suggest that C1s activated in degenerative cartilage matrix of RA but not in that of OA. C1s is thought to participate in the pathogenesis of RA through its collagenolytic activity in addition to the role in the classic cascade. PMID- 10364917 TI - CYP2C19 genotype does not represent a genetic predisposition in idiopathic systemic lupus erythematosus. AB - BACKGROUND: The aetiology of systemic lupus erythematosus (SLE) is still unknown. In several cases, however, chemicals or drugs were identified as aetiological agents and associations with certain phenotypes of drug metabolising enzymes have been reported. The purpose of this study was to discover if there is an association between CYP2C19 polymorphism and susceptibility to SLE. METHODS: Racemic mephenytoin (100 mg orally) was given to healthy volunteers (n = 161) and SLE patients (n = 37) and then S-mephenytoin and R-mephenytoin were determined in eight hour urine samples. A 10 ml blood sample was obtained from healthy volunteers (n = 80) and SLE patients (n = 69) for genotypic assay. Each blood sample was tested for the detection of CYP2C19*1 and CYP2C19*2 (formerly wt and m1 respectively) by oligonucleotide ligation assay. RESULTS: The ratio of S/R mephenytoin ranged from < 0.1 to 1.293 in healthy subjects and from < 0.1 to 1.067 in SLE patients. PM phenotype was observed in 2 of 37 patients with idiopathic SLE (5.4%) and 6 of 161 healthy subjects (3.7%). There were no significant differences in the frequency of PM phenotypes between the groups (Fisher's exact test, p = 0.64) or in the frequency distribution profiles of ratios of S-mephenytoin to R-mephenytoin. No significant differences in distribution of overall genotypes and in allele frequencies were observed between the two groups. No significant relation was found between clinical features and the overall genotype. CONCLUSION: The results of this study indicate that CYP2C19 genotype does not represent a genetic predisposition in idiopathic SLE patients. PMID- 10364918 TI - Examination of synovial fluid and serum hyaluronidase activity as a joint marker in rheumatoid arthritis and osteoarthritis patients (by zymography). AB - OBJECTIVE: Hyaluronic acid (HA) is an important joint marker and the substrate for hyaluronidase (HAase). Synovial fluid (SF) and serum HAase were measured to investigate the potential use of HAase as a joint marker in rheumatoid arthritis (RA) and osteoarthritis (OA) patients. METHODS: The subjects were 39 patients with RA and 42 patients with OA. HAase activity was measured by zymography and its relation with various parameters examined statistically. RESULTS: In RA SF a positive correlation (r = 0.458, p = 0.0186) was found between SF HAase activity and the concentration of serum C reactive protein. A positive correlation (r = 0.45, p = 0.024) was also found between SF HAase activity and platelet count in the RA group. Serum HAase activity in the RA group was significantly higher than in the OA group (p < 0.0001) and normal controls (p < 0.0001). CONCLUSION: The results suggest that SF HAase activity could be used as a marker of synovial inflammation. PMID- 10364919 TI - Circulating soluble adhesion molecules in patients with giant cell arteritis. Correlation between soluble intercellular adhesion molecule-1 (sICAM-1) concentrations and disease activity. AB - OBJECTIVE: To evaluate whether changes in concentrations of circulating adhesion molecules are related to disease activity in patients with giant cell arteritis (GCA). METHODS: A sandwich ELISA was used to measure soluble intercellular adhesion molecule-1 (sICAM-1), sICAM-3, vascular cell adhesion molecule-1 (sVCAM 1), E-selectin (sE-selectin), and L-selectin (sL-selectin) in serum and plasma samples from patients with GCA. A cross sectional study was performed on 64 GCA patients at different activity stages and on 35 age and sex matched healthy donors. Thirteen of these patients were evaluated at the time of diagnosis and serially during follow up. RESULTS: At the time of diagnosis, sICAM-1 concentrations were significantly higher in active GCA patients than in controls (mean (SD) 360.55 (129.78) ng/ml versus 243.25 (47.43) ng/ml, p < 0.001). In contrast, sICAM-3, sVCAM-1, sE-selectin, and sL-selectin values did not differ from those obtained in normal donors. With corticosteroid administration, a decrease in sICAM-1 concentrations was observed, reaching normal values when clinical remission was achieved (263.18 (92.7) ng/ml globally, 293.59 (108.39) ng/ml in the group of patients in recent remission, and 236.83 (70.02) ng/ml in those in long term remission). In the 13 patients followed up longitudinally, sICAM-1 values also normalised with clinical remission (225.87 (64.25) ng/ml in patients in recent remission, and 256.29 (75.15) ng/ml in those in long term remission). CONCLUSIONS: Circulating sICAM-1 concentrations clearly correlate with clinically apparent disease activity in GCA patients. Differences with results previously found in patients with other vasculitides may indicate that different pathogenic mechanisms contribute to vascular inflammation in different disorders. PMID- 10364920 TI - The A-G polymorphism in exon 1 of the CTLA-4 gene is not associated with systemic lupus erythematosus. AB - OBJECTIVES: Factors contributing to the development of systemic lupus erythematosus (SLE) remain largely unknown although are likely to include both environmental and genetic components. Studies on murine lupus have indicated a role for an antibody that blocks binding of cytotoxic T lymphocyte associated-4 (CTLA-4) to B7 on antigen presenting cells in the treatment of disease, suggesting that CTLA-4 may play an important part in the disease process. This study, therefore, investigated the frequency of a previously described A-G polymorphism in exon 1 of the CTLA-4 gene, the G allele of which has shown to be associated with both Graves' disease and type I diabetes, to determine whether this polymorphism was playing a part in the development of SLE. METHODS: One hundred and twenty six SLE patients and 363 control subjects were genotyped for the A-G polymorphism in exon 1 of the CTLA-4 gene. Target DNA was amplified using the polymerase chain reaction and the resulting product was digested using the BbvI restriction enzyme. RESULTS: No differences in allele or genotype frequencies were observed between patients with SLE and control subjects. CONCLUSION: These data suggest that the A-G polymorphism in exon 1 of the CTLA-4 gene does not play a part in the genetic susceptibility to the development of SLE. PMID- 10364921 TI - Effects of methotrexate on cartilage metabolism. PMID- 10364922 TI - Evidence-based surgery and the internet. AB - The preliminary version of the Evidence-Based Surgery (EBS) web pages was first released by the College Library in December 1998. In just four weeks there were over 1,600 recorded 'hits'. These draft pages were made known only to those surgeons and trainees participating on the various surgical e-mail discussion forums such as the Association of Surgeons in Training Mailing Lists. The popularity of the draft EBS pages and the encouraging feedback has rendered the on-going updating of the EBS pages a worthwhile activity. PMID- 10364923 TI - The college and the Paris Ecole de Chirurgie collaborate in surgical skills teaching. PMID- 10364924 TI - Report of the Working Party to Review Organ Transplantation. PMID- 10364925 TI - Ratio of basic surgical trainees to type 1 specialist registrar programmes 1999/2000/2001/2002. AB - It is recently becoming increasingly apparent that there is a significant discrepancy between the number of basic surgical trainees seeking Type 1 higher surgical training programmes and the number of those programmes available for appointment. PMID- 10364926 TI - East Anglian orthopaedic training survey. PMID- 10364927 TI - Anatomy, demonstrating and basic surgical training. AB - Calmanisation of surgical training has led to the introduction of the concept of a two-year basic surgical training (BST) rotation. Successful completion of this training is assessed by the new MRCS/AFRCS examination. Within these constraints there is no room for the previously popular anatomy demonstrating posts. The aim of this study, therefore, was to examine the views of FRCS and MRCS surgical trainees about their own demonstrating experience, their current anatomical knowledge and on the future value of anatomy demonstrating. PMID- 10364928 TI - Tackling the quality agenda in surgery: taking comparative audit into the next century. PMID- 10364929 TI - How today's registrar duties compare with yesteryear's... PMID- 10364930 TI - Assistants in surgical practice: a discussion document. PMID- 10364931 TI - Teleteaching--a practical and economical method of delivering surgical education. AB - The Plymouth Regional MRCS Course was initiated in 1997 to support the College STEP learning course and supplement the BST clinical training programmes. The course is a comprehensive fortnightly day-release scheme, covering, over one year in 21 full days of teaching, the entire MRCS syllabus in a structured programme of learning. PMID- 10364932 TI - Training in academic surgery. PMID- 10364933 TI - Consultant supervision of surgical trainees: an objective assessment of how much actually occurs. AB - Consultant-supervised operative experience must be at the core of any training programme. The level of consultant supervision of United Kingdom trainees is largely unknown. In this study, the unique Lothian Surgical Audit database was used to assess consultant supervised training. PMID- 10364934 TI - The specialist registrar puzzle: do all the pieces fit? AB - The review of medical training by the Working Party headed by Sir Kenneth Calman culminated in the introduction of the specialist registrar (SpR) training grade. Surgery and Radiology were the vanguard specialties, with the SpR post being introduced in December 1995. PMID- 10364935 TI - Should we have guidelines on trainees' research? PMID- 10364936 TI - Nationwide survey of basic surgical training and trainee satisfaction. PMID- 10364937 TI - Have you a problem with clinical audit? PMID- 10364938 TI - A cost analysis of day-stay surgery in otolaryngology. PMID- 10364939 TI - The effect of junior doctors' assistants on SHO workload. PMID- 10364940 TI - Would less time spent in training affect the quality of the training? PMID- 10364941 TI - Hugh Brendan Devlin CBE. PMID- 10364942 TI - Funding clinical research: the need for information and longer term strategies. AB - The Chief Medical Officer's Working Group on Specialist Medical Training recommended that training in research methodology should be a recognised component of all postgraduate training programmes and that further consideration be given by those responsible for postgraduate education, training and research to establishing how this might be achieved. Funding of the trainee in research is a crucial aspect of this directive, yet both trainers and trainees have described this as haphazard, invariably reliant on 'soft' money. The subject has raised wide discussion and debate. A questionnaire was sent to 205 consultant urologists in the UK, 154 (75%) replied and 130 (84%) had experience of research during their training. The first report examined their opinion about the contribution of research to their training; this report covers the questions directed towards funding, the source of their funding, whether sufficient funds, advice and information were available and where they might expect to obtain such details. The replies indicated a variety of sources of funding; knowledge about the financial support available for research was sparse and the majority considered there was insufficient advice and information available for trainees on the subject. Substantial funds are available for high quality scientific research programmes providing unprecedented opportunities for multidisciplinary collaboration that is essential for advancing clinical practice alongside technological developments. The process of obtaining support can be a time consuming exercise, raising the need for an administrative infrastructure to select, prioritise and co-ordinate an appropriate research strategy for the future. PMID- 10364943 TI - General surgeons and the management of head injuries. AB - Most head injuries in Great Britain and Ireland are managed either by orthopaedic or general surgeons. In response to growing anxiety about the arrangements for the management of head injuries, the Royal College of Surgeons of England late in 1997 set up a working party to report to the Senate of Surgery. The results of that survey are presented here. PMID- 10364944 TI - Pectoral myoplasty for recurrent pneumothorax: an extrathoracic solution to an intrathoracic problem. AB - The recurrence rate of spontaneous pneumothorax in patients with underlying lung disease can be as high as 50%. We present a novel method of treatment for recurrent pneumothorax based on the intrathoracic transfer of an extrathoracic muscle flap. PMID- 10364946 TI - The highs and lows of endovascular aneurysm repair: the first two years of the Eurostar Registry. AB - The Eurostar Registry was established in 1996 to collate information, from centres across Europe, on the outcome from endovascular grafting of aortic aneurysms. At the end of the first year of the project, data on 430 patients had been entered onto the database. In 420 patients (97.7%), the endografts were deployed without major complications. The 30-day mortality rate was low at 3.4% and deaths were confined mostly to 'high risk' patients with major co-morbidity. Endoleaks, which were present on discharge from hospital in 15.7% of patients, were associated with a significantly increased incidence of continued expansion of the aneurysm sac postoperatively (P < 0.01). Thus the early results confirmed the feasibility and low complication rate of endovascular repair of aneurysms, but a higher than expected incidence of endoleaks. At 2 years, 895 patients had been registered. The rate of early endoleaks remained significantly unchanged but another 18% of patients had developed new endoleaks during the first year of follow-up. Six delayed ruptures had been reported, 3 fatal. There were indications that 'self sealed' endoleaks continued to pressurise the aneurysm sac. Severe distortion of the grafts with kinking and other structural changes associated with postoperative longitudinal shrinking of the aneurysm sac was observed in 69% of patients at 1 year. Clinical complications associated with these changes included late endoleak and graft limb occlusion. Early unrealistic optimism about endovascular aneurysm repair has been replaced with a more realistic understanding of its benefits and limitations as a result of the Eurostar project and other registries. Randomised studies are now required to establish the most appropriate role for this approach, alongside established therapies. PMID- 10364945 TI - An audit of emergency abdominal aortic aneurysm repair to establish the necessity for an emergency vascular surgical rota. AB - Mortality for emergency abdominal aortic aneurysm (AAA) repair remains high but results of specialist vascular surgeons are superior to those of general surgeons. A retrospective audit was performed on all patients undergoing emergency AAA repair over 53 months at one hospital to determine the necessity for a vascular specialist on-call rota. Patients were stratified into two groups, those treated by specialist vascular surgeons and those treated by general surgeons. There were 37 patients in the vascular surgeon group and 36 in the general surgeon group. There was no significant difference between the two groups when age, sex distribution, APACHE II score on admission, pre-operative delay and type of rupture were considered. The average operating time was 114.7 min in the vascular surgeon group and 111.9 min in the general surgeon group. Total blood transfusion requirements, and postoperative duration of ventilation, inotrope therapy and intensive treatment unit stay were similar in the two groups. Intra operative, 30-day and cumulative hospital mortalities were 10.8% versus 8.3%, 32.4% versus 38.9% and 40.5% versus 38.9% in the vascular surgeon and general surgeon groups, respectively. The mortality figures compare favourably with other published series. As the results of the two groups were similar, there is currently no need for vascular surgeons to be routinely available for acute AAA surgery at our hospital. PMID- 10364947 TI - Treatment of infective and potentially infective complications of vascular bypass grafting using gentamicin with collagen sponge. AB - The use of gentamicin with collagen sponge (GCS) in treating patients with infective and potentially infective complications of vascular bypass grafting was reviewed. GCS was used in 25 patients. These were: 11 in situ applications to a proven graft infection, 2 to a superficial wound infection over an underlying graft, 4 at removal of an infected prosthesis, 4 for a persistent lymphatic leak, 3 for an anastomotic aneurysm and one for an anastomotic dehiscence. The GCS was used with rifampicin-soaked collagen impregnated Dacron in 3 of these patients. Of the 11 in situ treatments of a graft infection with GCS, 7 infections were successfully aborted, 3 grafts were removed resulting in 2 patients losing a leg and one patient died before the outcome was known. None of the other patients treated with GCS subsequently had infective sequelae. GCS is a simple, safe and often effective treatment when dealing with these difficult complications of arterial surgery, the use of which requires greater recognition. PMID- 10364948 TI - A prospective randomised trial of PIN versus conventional stripping in varicose vein surgery. AB - A prospective, randomised trial was carried out to examine the efficacy of perforate invagination (PIN, Credenhill Ltd, Derbyshire, UK) stripping of the long saphenous vein (LSV) in comparison to conventional stripping (Astratech AB, Sweden) in the surgical management of primary varicose veins. Eighty patients with primary varicosities secondary to sapheno-femoral junction (SFJ) incompetence and LSV reflux were recruited. Patients were randomised to PIN or conventional stripping with all other operative techniques remaining constant. Follow-up was performed at 1 and 6 weeks postoperatively. There were no statistically significant differences between the two techniques in terms of time taken to strip the vein, percentage of vein stripped or the area of bruising at 1 week. The size of the exit site was significantly smaller with the PIN device (P < or = 0.01). Optimal use of the conventional stripper provides results comparable to the PIN device. Choice of stripping device remains the surgeon's, bearing in mind that the PIN stripper achieves slightly better cosmesis. PMID- 10364949 TI - Pre-operative blood tests in children undergoing plastic surgery. AB - In a retrospective review of 1177 children presenting for plastic surgical procedures, investigations were performed in 487 and abnormal results were found in 138 as defined by variation from the local laboratory reference range. Most of the abnormalities were of no clinical significance. Twenty one children had abnormal haemoglobin results (the lowest was 9 g/dl) and 101 children had clinically insignificant platelet or white cell abnormalities. One child, with a family history of sickle cell trait, was confirmed as sickle-cell trait. No case was postponed as a result of these investigations. The non-selective ordering of pre-operative blood tests leads to unnecessary patient discomfort, the potential for additional superfluous investigations and higher costs. PMID- 10364950 TI - Paediatric day care surgery: a hidden burden for primary care? AB - The aims of this study were to identify prospectively 'concealed' postoperative problems, to assess the burden of paediatric day care surgery on primary care services and to define a normal recovery period for common children's procedures. At their first postoperative clinic visit, patients and parents were asked directly about postoperative problems, unscheduled contacts with the hospital or other health professionals, and the time taken to resume normal activity and return to school. A total of 651 children, median age 4 years (range 0-16 years) were included. Median time to the first clinic appointment was 42 days (range 4 235 days). There were 100 unscheduled postoperative contacts (15.5% of all episodes) of which 81 were with general practitioners. Most problems recorded were relatively minor, and most requiring major intervention were identified before the child left hospital. There was a very high incidence of wound related problems after circumcision. Most children were 'back to normal' within a few days (median 2 days, range 0-30 days) and back to school within a week (median 6 days, range 1-20 days). There was a high rate of primary care contact despite clear written and verbal advice given in hospital and the offer of open access for any concerns. Recovery from day care surgery was rapid and overall level of patient satisfaction was high. PMID- 10364952 TI - Allergic contact dermatitis to acrylates in disposable blue diathermy pads. AB - We report 2 cases of elicitation of allergic contact dermatitis to acrylates from disposable blue diathermy pads used on patients who underwent routine surgery. Their reactions were severe, and took approximately 5 weeks to resolve. Both patients gave a prior history of finger tip dermatitis following the use of artificial sculptured acrylic nails, which is a common, but poorly reported, cause of acrylate allergy. Patch testing subsequently confirmed allergies to multiple acrylates present in both the conducting gel of disposable blue diathermy pads, and artificial sculptured acrylic nails. We advocate careful history taking prior to surgery to avoid unnecessary exposure to acrylates in patients already sensitized. PMID- 10364951 TI - A prospective controlled trial of the efficacy of isopropyl alcohol wipes before venesection in surgical patients. AB - It has previously been suggested that skin preparation before venesection with antiseptic agents is unnecessary. However thousands of doctors and medical students continue to use isopropyl alcohol (IPA) swabs for venesection, at an estimated cost of 10,000 P per annum in a 500 bed hospital. An audit of IPA swab use among junior doctors and medical students at our institution was undertaken; 76% of doctors and 100% of medical students routinely prepared the skin with alcohol before venesection and only one used the swabs correctly. A randomised single-blind controlled trial was conducted of IPA versus no IPA skin preparation before venesection. There were 194 patients in the study, 93 in the IPA group and 101 controls. There was no statistical difference with respect to complications at the venepuncture site between the two groups. PMID- 10364953 TI - Transplant surgeons in training: is anybody out there? AB - There is a long-standing recognition that there is an organ donor shortage in the United Kingdom and Ireland (UK&E) that limits transplant activity. However, the fact that, at present, there are several unfilled consultant vacancies would suggest that a shortage of trained surgeons may soon be an equally important limiting factor. The aim of this current study was to identify all transplant trainees in the UK&E and to determine their career aspirations. A list of all trainees intending to practice as transplant surgeons was compiled. A combination of postal questionnaire and telephone interview was used to construct a database on past and present training in transplantation, and preferred type of consultancy was assessed both by direct questioning and by using a visual analogue scale to grade desirability of various posts. Of 110 potential trainees identified, 50 (45%) replied and indicated a desire to pursue a career in transplant surgery. Thirty-one intended practising in the UK&E (19 UK&E graduates and 12 overseas). The preferred consultancy (27/31) was transplantation (Tx) together with a second specialty while only four wanted a multivisceral practice. The mean score (0-10) for desirability of a multivisceral transplant post was 4.7, for renal transplant and vascular access it was 3.6 and for transplantation and a second specialty it was 8.4. We conclude that the majority of trainees do not wish to apply for pure transplant posts, either single organ or multivisceral, and that the majority wish to practice transplantation with a second specialty. In addition, there is still a major shortage of trainees and further studies are required to identify reasons why trainees fail to pursue a career in transplantation. PMID- 10364954 TI - The audit of orthopaedic trainee teaching improves supervision rates. AB - Inefficiency in surgical training has been identified as a result of low rates of supervision, with direct teaching of operating by consultants occurring in less than 20% of cases in papers dealing with general surgery and cardiac surgery training. The introduction of an audit system in an orthopaedic training programme was associated with an improvement of teaching from an already high 30% to 40% of cases. A logbook programme was introduced which allows easy analysis of the distribution of experience on an anatomical basis. PMID- 10364955 TI - Do obese patients bleed more? A prospective study of blood loss at total hip replacement. AB - This study compares blood loss at total hip replacement in obese and non-obese patients. We made a prospective study of intra-operative and postoperative blood loss in 80 consecutive primary cemented hip replacements. Patients' obesity was classified according to body mass index (BMI). Overall mean total blood loss was 1050 ml. Obese patients (BMI > 30) bled significantly more (P < 0.0001) than those of optimal weight (BMI < 26), whereas those overweight (BMI 26-30) did not. The mean excess blood loss in obese patients was 380 ml (95% confidence interval, 200-560 ml). At a time when the prevalence of obesity is increasing, this study quantifies the risks of greater blood loss with respect to obesity and aids informed consent. PMID- 10364956 TI - Posterior sternoclavicular dislocations--a diagnosis easily missed. AB - Posterior dislocation of the sternoclavicular joint is a relatively rare injury and can be difficult to diagnose acutely. We report 3 cases of posterior dislocation of the sternoclavicular joint who presented to the Accident & Emergency Department within a 3 month period. All 3 patients had sustained a significant injury to the shoulder region and complained of pain around the medial clavicle. Two patients had also complained of dysphagia following the injury. Plain X-rays of the shoulder and chest were reported as normal by junior and senior medical staff. The diagnosis was delayed until CT scans were performed, and once this was established, open reduction and stabilisation was performed. PMID- 10364957 TI - Adenocarcinoma within a rectal duplication cyst: case report and literature review. AB - Intestinal duplications are uncommon but recognised developmental anomalies. Duplications of the rectum are the most uncommon of these anomalies. They may present with perianal fistulae, bleeding, a pelvic mass or symptoms produced by a mass, or, rarely, malignant change. We present a case of an adenocarcinoma within a rectal duplication cyst which was initially thought to be inoperable but was treated by radical surgery. PMID- 10364959 TI - Square pegs in round holes: has psychometric testing a place in choosing a surgical career? A preliminary report of work in progress. AB - Methods of selection of candidates for training in surgery has long been regarded as lacking explicit criteria and objectivity. Our purpose was to discover the aptitudes and personality types of applicants for surgical posts at the outset, in order to discover which were most likely to result in a satisfactory progression through training and which were associated with career difficulties. This longitudinal predictive validation study has been undertaken in a London Teaching Hospital since 1994. After short-listing, but immediately before interview, all candidates for senior house officer posts in basic surgical training and in geriatric medicine were asked to undertake psychometric tests of numerical (GMA) and spatial (SIT7) reasoning, personality type (MBTI), and self rating of competency. There were no differences in ability scores between surgeons or geriatricians. Personality differences were revealed between the surgeons and the geriatricians, and between male and female surgeons. This study suggests that while there are no differences in ability between surgeons and geriatricians at the start of training, there are differences in personality. Long-term follow-up of the career development of this cohort of surgical SHOs is required to determine whether the psychometric measures described correlate with achievements of milestones in their surgical careers. PMID- 10364958 TI - Does screening for prostate cancer identify clinically important disease? AB - In this study the combination of digital rectal examination (DRE) and serum prostate-specific antigen (PSA) is shown to be effective for detecting early prostate cancer in a urological out-patient setting. PSA provides the means to detect cancer in men with normal DRE that may otherwise present as so-called incidental cancer at transurethral resection of the prostate (TURP) for apparently benign disease or later in the course of its natural history as locally advanced or metastatic disease. PSA progression in men with incidental cancer has been previously demonstrated to be predicted more reliably by residual cancer on needle biopsy after TURP than by tumour in the resected specimen and, therefore re-staging such patients is worthwhile when further treatment would be considered. Among men selected for radical prostatectomy, non-palpable tumours detected with PSA more predictable in pathological extent than incidental cancer and their particular pathological characteristics suggest they include clinically significant tumours that would progress if untreated to palpable and eventually metastatic disease. In view of this progressive behaviour, cancer detected by PSA should be considered clinically significant particularly in men with a life expectancy of at least 10 years. Therefore screening should be offered for such individuals, to detect and treat tumours at a curable stage and thereby eliminate the high mortality and often protracted morbidity commonly associated with metastatic disease. PMID- 10364961 TI - Recent advances in the management of cerebrovascular disease: the diminishing role of the surgeon? PMID- 10364960 TI - The biomechanics of leg ulceration. AB - Research performed in the late 1960s, using 24Na, suggested that the perfusion of skin and subcutaneous tissues is critically dependent on the relationship between capillary (Pc) and tissue pressures (Pt). Perfusion changes differed significantly between controls and patients with venous disease and the differences could be interpreted as evidence that Pt remained high in venous diseased patients. From this starting point, a biomechanical theory for the aetiology of venous ulceration was developed and tested by measuring skin elasticity, limb cross-sectional area and laser Doppler flux. The results confirm that, modelled as a two-compartment system (vascular and interstitial fluid), forces can be demonstrated sufficient to cause intermittent capillary closure and subsequent reperfusion injury. These forces are maximal in the gaiter area, the site of most leg ulcers. PMID- 10364962 TI - Angioplasty and stenting for atherosclerotic carotid artery disease. PMID- 10364963 TI - False aneurysm of the left ventricle after a stab wound to the chest. PMID- 10364964 TI - Detection of metachronous breast carcinoma: the role of follow-up? AB - Second primary (metachronous) breast carcinoma occurs at a rate of approximately 1% per year. Early detection of metachronous carcinomas will optimise the chances of curative treatment. The aim of this study was to identify the method of detection of metachronous carcinomas, so that efforts to detect these tumours can be made more focused. Thirteen patients presented twice to a surgical department in a 7-year period with second primary breast carcinomas. The means of detection of the second primary carcinoma was identified in each case. Eleven of the patients presented with new symptoms which they had noticed themselves. The remaining two carcinomas were detected mammographically, and their presence was confirmed on clinical examination. In no case was the second primary carcinoma detected by clinical examination alone. Metachronous carcinoma is unlikely to be detected by routine clinical examination, but rapid assessment of new symptoms should be facilitated. Follow-up mammography at regular intervals should also improve early detection of metachronous breast carcinoma. PMID- 10364965 TI - Colorectal cancer in adolescents. AB - Colorectal cancer, one of the most common malignancies among adults, is rare in adolescence. This low incidence coupled with non-specific symptoms and aggressive natural history leads to a poorer prognosis than in reported adult series. This article describes two cases of colorectal cancer in adolescents and reviews the literature regarding this rare condition. Earlier diagnosis and a greater understanding of the natural history may lead to improved treatment with concomitant improvements in survival. PMID- 10364966 TI - Controlled intraoperative water testing of left-sided colorectal anastomoses: are ileostomies avoidable? AB - Anastomotic leakage is a major problem in colorectal surgery, and previous studies have suggested that intraoperative identification of leaks allows repair at the time of surgery. This study examined whether testing allowed a defunctioning ileostomy to be safely omitted. A series of 102 consecutive patients underwent left-sided colorectal resection, 52 males and 50 females, mean age 65.7 years (range 16-89 years). After completion of the anastomosis, its integrity was tested by running saline into the rectum, using a manometer, to a maximum distending pressure of 30 cmH2O. Any leaks were repaired and the anastomosis retested. A defunctioning ileostomy was only performed if the anastomosis could not be shown to be leak-proof on testing. Patients underwent a contrast enema on the 8th postoperative day. Twenty-one (20.6%) patients failed the initial leakage test and 3 (3%) patients failed a second test. Two of these 21 patients went on to have a clinical leak, both of which were treated conservatively. Two defunctioning ileostomies were performed at the time of surgery. Sixteen (16.2%) had a leak on radiological testing, and there was clinical evidence of a leak in 5 (4.9%) patients. There were 3 (2.9%) deaths, but none of these had a leak on radiological testing. Incomplete anastomoses were successfully corrected intraoperatively. A defunctioning ileostomy was avoided in 98% of cases. Intraoperative testing to a pressure of 30 cmH2O is helpful in anterior resection, but does not guarantee that an intact anastomosis will remain intact postoperatively. PMID- 10364967 TI - Effect of prolonged infusion on vein calibre: a prospective study. AB - Infusion thrombophlebitis is common and is the principal limitation to intravenous nutrition (IVN) via a peripheral vein, yet its precise pathogenesis is unclear. Prospective observations were performed on patients in whom a hypertonic, acidic, nutritional emulsion was infused via fine-bore polyurethane catheters placed in peripheral veins. B mode ultrasound was used to determine vein calibre and proved to be a useful means for serial examination during intravenous infusion. Contrary to previous reports, no evidence of venospasm was observed. It is suggested that previous evidence of venoconstriction is erroneous and that other mechanisms are responsible for thrombophlebitis. PMID- 10364968 TI - Midline laparotomy incision: a technique with a different slant. PMID- 10364969 TI - Sucking injury or gas gangrene? PMID- 10364970 TI - Handwashing: simple, but effective. AB - Using ward rounds in the department of surgery at a major teaching hospital, and with the help of the preregistration house officers (PRHO), we assessed whether the lesson taught to us by Semmelweis had been forgotten. We asked the PHROs to count the number of patients examined by their consultant or registrar on a ward round, together with the number of wounds examined, and the number of times they washed their hands between patients. Over a 2-week period, following seven consultants and four registrars, 26 ward rounds were followed. Of 239 patient events, which are defined as a clinician reviewing a patient in order to assess their treatment, a total of 88 involved an examination (37%) and, of these, 41 had postoperative wounds (47%). The number of times clinicians washed their hands between examinations was 36 (41%). Between the two groups of clinicians, the consultants washed their hands 30 times in 55 examinations (55%), while the registrars washed their hands six times in 23 examinations (26%). When Semmelweis died in 1865 his beliefs were still largely ignored by clinicians. It would seem from our results that in both senior and junior staff the simple exercise of handwashing is not practised de rigor. For the safety of the patient and the clinician we recommend a more fastidious adoption of the handwashing practice. PMID- 10364971 TI - Impact of third molar removal on demands for postoperative care and job disruption: does anaesthetic choice make a difference? AB - A prospective cohort study was undertaken to investigate the influences of anaesthetic modality and surgical difficulty on social reintegration and demands on health services after third molar removal. The study was undertaken at the Oral and Maxillofacial Surgery Department, Cardiff Dental Hospital. Of 444 patients, 266 (60%) had their third molars removed. The main outcome measures included anaesthetic modality, surgical difficulty (WHARFE scores), utilisation of health services, effects on work, school and home life. In all, 101 (40%) patients were treated under local anaesthesia (LA) +/- intravenous (i.v.) sedation and 165 (60%) under general anaesthesia (GA); 81 (49%) as inpatients and 84 (51%) as day cases. Of these patients, 38 (14%) returned to the hospital and 74 (28%) utilised primary care services postoperatively in addition to a standard review appointment. Patients treated under GA made more demands on primary care services (chi 2 = 6.41, df = 2, P < 0.05) and took more time away from work (P < 0.05). Patients underestimated the time they needed to recover. There was similar disruption to job, college and home life. There were no links between disruption and particular anaesthetic modalities and surgical difficulty. Surgery under GA was linked to increased postoperative demands on primary care, but not secondary care, and to longer job disruption. This could not fully be attributed to surgical difficulty. PMID- 10364972 TI - Core cutter for harvesting cortical bone grafts for reconstructions of the ossicular chain. AB - Cortical bone autografts have been used to reconstruct the ossicular chain for more than 30 years. We describe a core cutter burr which facilitates the rapid harvesting of grafts which are suitable for a number of different types of reconstruction. The use of these grafts to reconstruct different defects of the ossicular chain is also presented. PMID- 10364973 TI - Acetabular augmentation for the treatment of unstable total hip arthroplasties. AB - Twenty-eight unstable total hip arthroplasties were treated with an acetabular augmentation wedge. Of the hips, 23 have had no further dislocations at a mean follow-up of 26 months. Five patients continued to dislocate and have needed further surgery. To our knowledge this is the largest reported series of acetabular augmentation with as good results as those of the most successful reported series of this technique, and a success rate comparable to other methods of treating recurrent dislocation. Careful patient selection, and using a thin augmentation wedge to avoid impingement, are important to the success of a technique which is a useful option in the management of recurrent dislocation. PMID- 10364974 TI - An audit of the investigation and treatment of localised prostatic cancer in the south west region. AB - Prostate cancer constitutes a major health care dilemma in terms of treatment options available and increasing patient load on both a regional and national level. An audit was undertaken of all patients in the South West Region with localised prostate cancer newly diagnosed in 1993 to assess regional management of this disease. In 1993, 1407 patients were newly diagnosed as having prostatic cancer. Patients > 75 years old and those with a prostate-specific antigen (PSA) > 40 ng/ml were excluded, leaving 262 patients whose case notes were examined. The interval between referral and clinic (mean 67 days) was altered by the presence of a GP performed PSA, being shorter if the PSA was > 10 ng/ml (average 54 days) than if the PSA was < 10 ng/ml (average 104 days). Overall, 34% of patients underwent radical treatment (10% radical prostatectomy and 24% radiotherapy). In all, 27% received hormone manipulation or orchidectomy, and the remainder 'watchful waiting'. The majority (78%) of patients < 60 years old received radical treatment, as did 35% of those 60-70 years and 15% of 70-75 year olds. Over 90% of tumours were category T1 and were well or moderately differentiated. All patients had a histological diagnosis and 84% had their tumour staged before treatment. This study highlighted the need for improvements in patient assessment, improved note keeping and a regional cancer register to allow ongoing assessment of patient management. This audit of management of localised prostate cancer serves as a baseline from which to initiate and monitor improvements in the service regionally and will also allow assessment of the impact of such changes. PMID- 10364975 TI - Splenic metastasis from a rectal tumour: an unusual presentation. PMID- 10364976 TI - Iatrogenic oesophageal perforations: a clinical review. PMID- 10364977 TI - The continuing challenge of parastomal hernia: failure of a novel polypropylene mesh repair. PMID- 10364978 TI - Changing patterns of treatment for chronic anal fissure. PMID- 10364980 TI - Endoscopic sinus surgery: are junior doctors being properly trained? PMID- 10364979 TI - Mortality after elective abdominal aortic aneurysm repair: not where ... but how many and by whom. PMID- 10364981 TI - Simple technique for non-operative removal of ureteric stents after renal transplantation. PMID- 10364982 TI - Labial fat pad grafts (modified Martius graft) in complex perianal fistulas. PMID- 10364983 TI - Securing of surgical drains in the neck. PMID- 10364984 TI - Securing of surgical drains in the neck. PMID- 10364985 TI - Intraoperative conservation of red cell mass: controlled hypotension or haemodilution--not necessarily mutually exclusive? PMID- 10364986 TI - Central nerve block and thromboprophylaxis--is there a problem? PMID- 10364987 TI - Propofol and epilepsy. PMID- 10364988 TI - Acute normovolaemic haemodilution vs controlled hypotension for reducing the use of allogeneic blood in patients undergoing radical prostatectomy. AB - Blood loss in patients undergoing radical prostatectomy may be substantial. In a randomized, prospective study, we assessed two methods of reducing the need for allogeneic blood transfusion with regard to efficacy and costs. Sixty patients undergoing retropubic radical prostatectomy were allocated randomly to one of three groups. In group 1 (n = 20), acute normovolaemic haemodilution (ANH) was initiated after induction of anaesthesia; autologous blood 15 ml kg-1 was withdrawn and replaced by colloid solutions (gelatin) to maintain haemodynamic stability. In group 2 (n = 20), controlled hypotension was established using sodium nitroprusside (target mean arterial pressure (MAP) approximately 50 mm Hg). Group 3 (n = 20), without manipulations, served as a control group. Troponin T (TnT), a sensitive marker for myocardial ischaemia, and various coagulation variables were measured in the perioperative period. Packed red blood cells (PRBC) were given when haemoglobin concentration was less than 7 g dl-1. Cost calculations did not include hospital overhead costs or staff costs. In the ANH group, mean 1278 (SD 150) ml of autologous blood were withdrawn. Significantly more volume was infused in the ANH patients (gelatin 2450 (550) ml) than in the two other groups. Coagulation data (platelet count, activated partial thromboplastin time (aPTT), fibrinogen, antithrombin III (AT III), D-dimers) did not differ significantly between the three groups. The hypotension group had significantly lower blood loss (1260 (570) ml), whereas the ANH (1820 (680) ml) and control group (1920 (590) ml) did not differ significantly. Patients in the hypotension group needed significantly less PRBC (total 14 units; 75% of patients did not need PRBC) than the ANH (total 21 units; 55% of patients did not need PRBC) or control patients (total 28 units; 40% of patients did not need PRBC). Total costs were lowest in the hypotension group (41% less than in the control patients) (P < 0.05). We conclude that the use of hypotension during radical prostatectomy resulted in approximately 40% reduction in total transfusion costs. ANH was less effective and more costly than controlled hypotension. PMID- 10364989 TI - Thrombelastogram reveals hypercoagulability after administration of gelatin solution. AB - We have compared the effects of gelatin, low molecular weight hydroxyethyl starch (HES) or albumin on tests of haemostasis and on the thrombelastogram in 42 ASA I patients undergoing total hip or knee replacement. Patients were allocated randomly to receive one of the three blood substitutes to obtain moderate intraoperative haemodilution. Blood loss and packed red cell infusion was the same in each group. A greater amount of gelatin was given (1.5 times the measured blood loss) because of its shorter half-life. There was a statistically significant but clinically negligible decrease in platelets count, prothrombin time and fibrinogen, and an increase in bleeding time in all groups. Platelets were slightly but significantly lower after HES. Haemodilution was comparable between groups. TEG showed a state of hypercoagulability in the gelatin group with a significant decrease in r, r + k and an increase in alpha angle. PMID- 10364990 TI - Pulse oximetry plethysmographic waveform during changes in blood volume. AB - Systolic pressure variation (SPV) and its dDown component have been shown to be sensitive factors in estimating intravascular volume in patients undergoing mechanical ventilation. In this study, ventilation-induced changes in pulse oximeter plethysmographic waveform were evaluated after removal and after reinfusion of 10% estimated blood volume. The plethysmographic waveform variation (SPVplet) was measured as the difference between maximal and minimal peaks of waveform during the ventilatory cycle, and expressed as a percentage of the signal amplitude during apnoea. dUp(plet) and dDown(plet) were measured as the distance between the apnoeic plateau and the maximal or minimal plethysmographic systolic waveform, respectively. Intravascular volume was changed by removal of 10% of estimated blood volume and followed by equal volume replacement with Haemaccel. A 10% decrease in blood volume increased SPVplet from mean 17.0 (SD 11.8)% to 31.6 (28.0)% (P = 0.005) and dDown(plet) from 8.7 (5.1)% to 20.5 (12.9)% (P = 0.0005) compared with baseline. Changes in plethysmographic waveform correlated with changes in arterial SPV and dDown (r = 0.85; P = 0.0009). In the absence of invasive arterial pressure monitoring, ventilation-induced waveform variability of the plethysmographic signal measured from pulse oximetry is a useful tool in the detection of mild hypovolaemia. PMID- 10364991 TI - Formation of nitrogen dioxide from nitric oxide and their measurement in clinically relevant circumstances. AB - Therapy with inhaled nitric oxide in oxygen requires adequate monitoring of nitric oxide and nitrogen dioxide. The characteristics of chemiluminescence and electrochemical measurement techniques were determined by analysis of continuously flowing gas mixtures and comparisons with traceable gas standards. Gas mixtures were also diluted with mass flow controllers and in addition created in ventilator breathing systems. Factors influencing the formation of nitrogen dioxide were defined. Both techniques accurately measured nitric oxide (10-80 parts per million, ppm) and nitrogen dioxide (0.5-5 ppm) in normoxic and hyperoxic (90% oxygen) gas in the studied ranges. Nitrogen dioxide in hyperoxic gas had three origins: (1) from the premixing point of nitric oxide in nitrogen, (2) as a result of the mixing process, and (3) from post-mixing and time dependent continuous formation of nitrogen dioxide in oxygen. We conclude that adequate monitoring is possible and that factors affecting nitrogen dioxide generation can be defined. PMID- 10364992 TI - Dry soda lime markedly degrades sevoflurane during simulated inhalation induction. AB - We have investigated gas composition during simulated inhalation induction with sevoflurane to elucidate possible mechanisms of incidental prolonged induction times and airway irritation. Using a circle system, 8% sevoflurane in oxygen 6 litre min-1 was washed into an absorbing canister filled with fresh soda lime containing 2.9% KOH (Draegersorb, 'D') or no KOH (< 0.01%, Sofnolime, 'S'). The absorbent was dried by oxygen 20,000 litre before every second experiment. Maximum soda lime temperatures attained after 4-6 min were 107 degrees C using dry D and 62 degrees C (61 degrees C) with dry S. Temperature did not increase with fresh soda lime. With dry soda lime, sevoflurane was not detected at the T piece for 3 min and reached 6-7% within 6-10 min. After 1 min, we detected methanol and compound A (CH2F-O-C(= CF2) (CF3)). Total amounts over 20 min were: methanol 1125 mg (D dry), 334 mg (S dry) and < 5 mg (fresh soda lime); compound A 148 mg (D dry), 13 mg (S dry) and 3-8 mg (fresh); and fluoride 8.5 mg (D dry), 3.3 mg (S dry) and 1 mg (fresh). Formaldehyde was detected only with dry lime (D > 2.5 mg, S > 0.6 mg). In summary, the use of moist soda lime is of crucial importance during inhalation induction. With dry soda lime, the patient may inhale potentially toxic degradation products in significant amounts. Sevoflurane degradation is aggravated by a high KOH content of the lime. The observed airway irritation may be caused by formic acid, which is generated in isomolar concentrations with methanol (Cannizzaro reaction). The amount of compound A found with dry KOH-containing lime is unlikely to be noxious. PMID- 10364993 TI - Effect of propofol on the electrocorticogram in epileptic patients undergoing cortical resection. AB - We have compared the effect of clinical doses of propofol with thiopental on epileptiform activity in the electrocorticograms (ECoG) of 20 epileptic patients undergoing temporal lobe resection. After baseline ECoG had been obtained, with inspired concentrations of 0.5-1% isoflurane and 70% nitrous oxide to provide background anaesthesia, subjects were allocated randomly to receive boluses of either thiopental 25 mg or propofol 20 mg i.v. every 30 s to a maximum of 5 mg kg 1 or until burst suppression was seen. The ECoG was recorded throughout administration and for 10 min thereafter. After return of baseline ECoG tracings, the alternate agent was administered. The amount of epileptiform activity was recorded on an ordinal rating scale, an increase being indicated by either a rise of at least one category on the scale or discharges occurring at a minimum of one new site. Activation occurred more frequently with thiopental but the difference was not significant. This study suggests that propofol has no greater proconvulsive effect than thiopental, a drug commonly used in managing status epilepticus. PMID- 10364994 TI - Prediction of movement at laryngeal mask airway insertion: comparison of auditory evoked potential index, bispectral index, spectral edge frequency and median frequency. AB - We have studied 46 patients to compare the efficacy of the auditory evoked potential (AEP) index, bispectral index (BIS), 95% spectral edge frequency (SEF) and median frequency (MF) in predicting movement in response to insertion of the laryngeal mask airway (LMA). Anaesthesia was induced with target-controlled infusions of propofol and alfentanil. After loss of eyelash reflex and adequate jaw relaxation, the LMA was inserted without the assistance of a laryngoscope or neuromuscular blocker. Patients who showed any visible spontaneous muscle movement within 1 min of LMA insertion were defined as movers. Values in movers and non-movers at 30 s before LMA insertion were analysed. Only AEP index discriminated between movers and non-movers with a prediction probability of 0.872. BIS, SEF and MF could not predict movement at LMA insertion. AEP index was the most reliable predictor of movement in response to LMA insertion. PMID- 10364996 TI - Small tidal volume ventilation using a zero deadspace tracheal tube. AB - The zero deadspace tracheal tube (ZEDS-TT) is a double-lumen endobronchial tube with a truncated bronchial limb. Functionally it is unrelated to the familiar endobronchial tube used in lung isolation surgery. It is placed in the same position as a regular tracheal tube and, by means of special connectors, one limb is used for inspiration and the other for expiration, thereby greatly reducing anatomical and apparatus deadspace. In this study, we have compared respiratory and ventilatory effects of reduction of tidal volume (VT) via a single-lumen tracheal tube and the ZEDS-TT during controlled ventilation with a Siemens Elema 900C Servo ventilator. Eleven consenting adult patients (ASA I and II) undergoing elective peripheral surgery were studied. Starting at a VT value of 10 ml kg-1, data were recorded for each tube type. VT was reduced by 2.5 ml kg-1 every 10 min and stabilized data recorded. Minute volume was kept constant by increasing ventilatory frequency at each reduction in VT. We found that the ZEDS-TT produced a significant reduction in PaCO2 and airway pressure for any VT used, while maintaining oxygenation. PMID- 10364995 TI - Placebo-controlled study of inhaled nitric oxide to treat hypoxaemia during one lung ventilation. AB - The aim of this prospective, placebo-controlled study was to assess if unilaterally inhaled nitric oxide 20 ppm could treat hypoxaemia during one-lung ventilation. Sixty patients undergoing pulmonary resection using a lateral thoracotomy were allocated randomly to a control or nitric oxide group (NO group). During one-lung ventilation in the lateral decubitus position, the lungs were ventilated mechanically with 90% oxygen--10% nitrogen. After randomization, if PaO2 decreased to less than 9.3 kPa during one-lung ventilation, nitric oxide 20 ppm or nitrogen was added to the inspired gas. The criterion for treatment efficacy was an increase in PaO2 to greater than 9.3 kPa after gas administration. Eight patients in the control group and eight in group NO experienced hypoxaemia during one-lung ventilation. PaO2 was not significantly different in the two groups at the time of gas administration (control group mean 8.0 (SD 0.6) kPa; NO group 8.5 (0.5) kPa). The efficacy criterion was reached in two of eight patients in the control and NO groups. The results of this study showed that inhaled nitric oxide 20 ppm, administered in the dependent lung, was not superior to nitrogen in the treatment of hypoxaemia during one-lung ventilation. Nitric oxide should not be recommended as an alternative to conventional management of hypoxaemia in this condition. PMID- 10364997 TI - Respiratory effects of low-dose bupivacaine interscalene block. AB - In this double-blind study, interscalene brachial plexus (ISBP) block was performed in 11 volunteers using 10 ml of either 0.25% (n = 6) or 0.5% (n = 5) bupivacaine with epinephrine 1:200,000. Diaphragmatic excursion, respiratory function and neural function were assessed for 90 min. Our results showed that hemidiaphragmatic excursion declined significantly after block in the 0.5% group and paradoxical movement during inspiration was more common than in the 0.25% group. Forced vital capacity and forced expiratory volume in 1 s declined significantly in the 0.5% group (mean 74.6 (SD 13.0)% and 78.2 (19.9)% of baseline, respectively) but not in the 0.25% group. Sensory anaesthesia in the upper limb was found consistently in both groups, although biceps paralysis occurred earlier after 0.5% bupivacaine. We conclude that ISBP block using 10 ml of 0.25% bupivacaine provided upper limb anaesthesia to pinprick in C5-6 dermatomes with only occasional interference with respiratory function. PMID- 10364998 TI - I.v. diclofenac and ketorolac for pain after thoracoscopic surgery. AB - We studied intensity of pain, cumulative morphine consumption, ventilatory and renal function, and haemostasis in patients undergoing video-assisted thoracoscopic surgery and receiving a 2-day i.v. infusion of diclofenac, ketorolac or saline. Plasma concentrations of the two NSAID were also measured. The study was randomized, double-blind and placebo-controlled, with 10 patients in each group. Patients experienced mainly moderate pain. Mean consumption of i.v. morphine during the first day after operation was 57 (SEM 11) mg in the placebo group. Diclofenac and ketorolac were equally effective in reducing total morphine consumption (61% and 52%, respectively). Adverse events were similar and minor. Greater variability in plasma concentrations of ketorolac were detected compared with diclofenac. PMID- 10364999 TI - Comparison of intrathecal and epidural diamorphine for elective caesarean section using a combined spinal-epidural technique. AB - To assess calculated equivalent doses of intrathecal and epidural opioids for elective Caesarean section in terms of quality and duration of analgesia, and incidence of side effects, we have compared 50 patients, allocated randomly to one of two groups to receive either diamorphine 0.25 mg intrathecally (group 1) or 5 mg epidurally (group 2), in addition to intrathecal bupivacaine 10 mg, using a combined spinal-epidural technique. There was no significant difference in duration of analgesia between groups (group 1 mean 14.6 (SD 5.9) h, group 2 14.2 (6.5) h; mean difference 0.8 h; 95% Cl -2.8-4.5; P = 0.65) or quality of analgesia (VAPS and VRS scores). The degree of pruritus was similar in both groups (80-88%) but the incidence of postoperative nausea and vomiting was significantly higher in the epidural group (24% vs 4%; P < 0.05). Intrathecal diamorphine 0.25 mg produced the same duration and quality of postoperative analgesia as epidural diamorphine 5 mg for elective Caesarean section but with significantly less nausea and vomiting. PMID- 10365000 TI - Comparison of midwife top-ups, continuous infusion and patient-controlled epidural analgesia for maintaining mobility after a low-dose combined spinal epidural. AB - We studied 133 women given a combined spinal-epidural for analgesia in labour. The initial intrathecal dose contained bupivacaine 2.5 mg with fentanyl 25 micrograms. When the mothers were comfortable, they were allocated randomly to one of three groups: continuous infusion (group Cl, n = 46), midwife top-ups (group MW, n = 43) or patient-controlled epidural analgesia (group PCEA, n = 44), to maintain analgesia throughout labour. All epidural solutions contained 0.1% bupivacaine and fentanyl 2 micrograms ml-1. Motor block was assessed by the mother's ability to straight leg raise (SLR). Four hours after combined spinal epidural analgesia, 88.1% of women could SLR in group MW, 83.7% in group PCEA and 57.8% in group Cl (P = 0.002). Total use of bupivacaine was highest in group Cl (mean 11.3 (SD 3.3) mg h-1) compared with group MW (7.5 (3.1) mg h-1) and group PCEA (9.1 (2.1) mg h-1) (P < 0.001). Analgesia was similar between groups and overall satisfaction was equally high. PMID- 10365001 TI - Cerebral response to haemodilution during cardiopulmonary bypass in dogs: the role of nitric oxide synthase. AB - During cardiopulmonary bypass, haemodilution is standard practice and is accompanied by increases in cerebral blood flow (CBF). We investigated if changes in cerebral vascular resistance (CVR) during cardiopulmonary bypass-haemodilution are dependent on nitric oxide synthase. The cerebral response to haemodilution in nine dogs treated with the nitric oxide synthase inhibitor, N omega-nitro-L arginine methyl ester (L-NAME), was compared with a control group (n = 8). Both groups underwent serial isovolaemic haemodilution (target packed cell volumes 0.39, 0.26, 0.19 and 0.14) using 6% dextran 70. CBF, CVR and cerebral metabolic rate for oxygen (CMRO2) were measured. While initial CVR was different in the two groups, haemodilution-dependent reductions in CVR were equivalent and the curves describing the packed cell volume-CVR relationship were parallel in control and nitric oxide synthase inhibition groups. Our data indicate that nitric oxide synthase does not play a primary role in the cerebral response to haemodilution. PMID- 10365002 TI - Isoflurane can indirectly depress lumbar dorsal horn activity in the goat via action within the brain. AB - We have examined the response of lumbar dorsal horn cells to a noxious mechanical stimulus during differential delivery of isoflurane to the brain and spinal cord of goats. We hypothesized that isoflurane, acting in the brain, would depress dorsal horn neuronal responses to a noxious mechanical stimulus applied to the hindlimb. Eight goats were anaesthetized with isoflurane and neck dissections performed which allowed cranial bypass. Lumbar laminectomies were performed to allow measurements of single-unit dorsal horn neuronal activity. Isoflurane 1.3% was administered before bypass, and during differential delivery it was administered at each of the following head/torso combinations: 1.3%/1.3%, 0.8%/1.3%, 0.3%/1.3%, 1.3%/0.8%, 0.8%/0.8% and 0.3%/0.8%. When the torso isoflurane concentration was 1.3%, decreasing cranial isoflurane from 1.3% to 0.3% did not significantly affect dorsal horn responses (from mean 325 (SD 262) to 379 (412) impulses min-1; P < 0.05). However, when torso isoflurane was 0.8%, decreasing cranial isoflurane from 1.3% to 0.3% increased mean evoked dorsal horn activity by 42% (388 (359) to 551 (452) impulses min-1; P < 0.05). These data suggest that the major effect of isoflurane on dorsal horn responses to noxious stimuli is direct, but there is an indirect effect occurring via descending projections from supraspinal regions. PMID- 10365003 TI - Glycine toxicity after high-dose i.v. infusion of 1.5% glycine in the mouse. AB - Glycine 1.5% is the most widely used irrigating fluid during endoscopic procedures. To investigate if glycine toxicity is a mechanism promoting a fatal outcome when the solution is absorbed, we administered glycine 200 or 300 ml kg-1 dissolved in sterile water or normal saline, and also normal saline alone, over 60 min by i.v. infusion to 100 mice under methoxyflurane anaesthesia. Survival rates were 29% after 1.5% glycine, 21% after 1.5% glycine in normal saline, 67% after normal saline and 100% in controls. Both solutions containing glycine induced bradycardia and prolongation of the PQ interval and QRS duration, while only 1.5% glycine increased the water content of the myocardium. These results suggest that glycine promotes bradycardia and death, regardless of whether hyponatraemia or hypo-osmolality is present. We conclude that glycine toxicity is an important factor that increases the risk of administration of an irrigating fluid. PMID- 10365004 TI - Drugs and sex differences: a review of drugs relating to anaesthesia. PMID- 10365005 TI - Time course of neurone-specific enolase and S-100 protein release during and after coronary artery bypass grafting. AB - Serum neurone-specific enolase (NSE) and S-100 protein are well established as markers of cerebral injury, and have been used as markers of neuronal and glial cell damage, respectively, after cardiac surgery with cardiopulmonary bypass (CPB), but the speed of their increase during CPB has not been studied. Therefore, we have investigated the time course of NSE and S-100 release during and after CPB. We studied 18 adult patients undergoing elective coronary artery bypass grafting (CABG). Standard hypothermic (32 degrees C) pulsatile bypass with membrane oxygenation was used. Blood samples were obtained at induction, before bypass, before rewarming, at the end of rewarming, 10 min, 1 h and 8 h after bypass and 1, 2 and 3 days after surgery. NSE and S-100 were assayed using immunoradiometric assay kits (Sangtec Medical). NSE and S-100 release followed similar time courses. Both increased sharply during bypass, reached peak concentrations at the end of rewarming (mean 25.55 (SEM 2.79) and 1.65 (0.23) microgram litre-1, respectively), had decreased significantly by the end of operation and returned to pre-bypass concentrations by the second day after surgery. No patient developed a major neurological deficit. When using NSE and S 100 assays to study cerebral dysfunction in relation to CPB, postoperative samples miss peak (end-bypass) concentrations, and studies should be designed to include intraoperative samples. PMID- 10365006 TI - Sister chromatid exchange in human lymphocytes exposed to isoflurane and nitrous oxide in vitro. AB - The question of whether or not inhalation anaesthetics are genotoxic remains controversial. Therefore, we have studied the in vitro genotoxic potential of isoflurane and nitrous oxide in human lymphocytes. Blood samples were obtained from eight healthy male, non-smoking volunteers, which were incubated and exposed to increasing concentrations of isoflurane (0.0, 0.3, 0.6 and 1.2 mmol litre-1) or 50% nitrous oxide in oxygen. Baseline sister chromatid exchange (SCE) rate per cell was mean 7.65 (SD 1.5) which increased to 9.15 (1.0), 9.55 (1.4) and 9.95 (1.8) SCE/cell during exposure to isoflurane 0.3, 0.6 and 1.2 mmol litre-1, respectively. During 50% nitrous oxide exposure, SCE rate was 9.26 (1.4). The difference between the control and exposed cells was statistically significant (P < or = 0.05). We conclude that exposure to nitrous oxide and subanaesthetic concentrations of isoflurane can produce genetic damage in peripheral lymphocytes in vitro. PMID- 10365007 TI - Dissociation of pituitary-adrenal and catecholamine activation after induced cardiac arrest and defibrillation. AB - To avoid factors which confound attempts to characterize the neuroendocrine response to cardiac arrest, we studied the pituitary-adrenocortical and catecholamine responses to induced ventricular fibrillation (VF) and direct current cardioversion in 10 patients undergoing testing of 'implanted cardioverter defibrillator' devices under sedation. Plasma concentrations of epinephrine were increased 5 min after VF (from a mean basal of 0.39 (S.E.M. 0.09) to a peak of 0.632 (0.212) nmol litre-1; P < 0.05) but were unchanged at other times. Plasma concentrations of norepinephrine did not change at any time. Plasma concentrations of cortisol increased significantly at 10 min (from a mean of 367 (SEM 62) to 539 (64) nmol litre-1; P < 0.001) and remained increased 30 min after VF (470 (74) nmol litre-1; P < 0.05) but had returned towards baseline at 60 min, whereas plasma prolactin concentrations were increased at 5 min (from a mean of 224 (SEM 54) to 320 (63) mu. litre-1; P < 0.01) and remained increased until the end of the sampling period at 60 min (288 (65) mu. litre-1; P < 0.05). Concentrations of adrenocorticotrophic hormone (ACTH) (n = 5) tended to increase but this was not statistically significant. We conclude that a short period of cardiac arrest in lightly sedated humans activated the pituitary-adrenocortical axis but did not appear to stimulate catecholamine secretion. These findings raise questions about the nature and mechanisms of the neuroendocrine response to cardiac arrest. PMID- 10365008 TI - Antiemetic activity of the NK1 receptor antagonist GR205171 in the treatment of established postoperative nausea and vomiting after major gynaecological surgery. AB - In this double-blind, randomized, parallel group study, we have investigated the antiemetic activity of the potent and selective NK1 receptor antagonist GR205171 25 mg i.v. compared with placebo in the treatment of established postoperative nausea and vomiting (PONV) in patients after major gynaecological surgery performed under general anaesthesia. The incidence of PONV in the study population was 65%. Thirty-six patients were treated with placebo or GR205171 (18 patients per group). GR205171 produced greater control of PONV than placebo over the 24-h assessment period. The stimuli for emesis after PONV are multifactorial and the efficacy of GR205171 in this study supports the broad spectrum potential for NK1 receptor antagonists in the management of postoperative emesis. GR205171 was well tolerated and no adverse events were reported that would preclude the further development of this agent. PMID- 10365009 TI - Glycopyrrolate reduces nausea during spinal anaesthesia for caesarean section without affecting neonatal outcome. AB - We have tested the hypotheses that glycopyrrolate, administered immediately before induction of subarachnoid anaesthesia for elective Caesarean section, reduces the incidence and severity of nausea, with no adverse effects on neonatal Apgar scores, in a double-blind, randomized, controlled study. Fifty women received either glycopyrrolate 200 micrograms or saline (placebo) i.v. during fluid preload, before induction of spinal anaesthesia with 2.5 ml of 0.5% isobaric bupivacaine. Patients were questioned directly regarding nausea at 3-min intervals throughout operation and asked to report symptoms as they arose. The severity of nausea was assessed using a verbal scoring system and was treated with increments of i.v. ephedrine and fluids. Patients in the group pretreated with glycopyrrolate reported a reduction in the frequency (P = 0.02) and severity (P = 0.03) of nausea. Glycopyrrolate also reduced the severity of hypotension, as evidenced by reduced ephedrine requirements (P = 0.02). There were no differences in neonatal Apgar scores between groups. PMID- 10365010 TI - Levobupivacaine vs bupivacaine as infiltration anaesthesia in inguinal herniorrhaphy. AB - We have compared the anaesthetic and analgesic efficacy of levobupivacaine with that of racemic bupivacaine in 66 male patients undergoing ambulatory primary inguinal herniorrhaphy. Patients were allocated randomly in a double-blind manner to local infiltration anaesthesia (0.25% w/v 50 ml) with either racemic bupivacaine (n = 33) or levobupivacaine (n = 33). Scores for intraoperative pain and satisfaction with anaesthesia were recorded, together with perception of postoperative pain and need for supplementary postoperative analgesic medications in the first 48 h after operation. Intraoperative satisfaction with the infiltration anaesthesia was similar, with median scores of 77 (levobupivacaine) and 80 (bupivacaine) (VAS; 100 mm = extremely satisfied). Time averaged postoperative pain scores (48 h) were 8 (levobupivacaine) and 10 (bupivacaine) in the supine position, 13 (levobupivacaine) and 12 (bupivacaine) while rising from the supine position to sitting, and 9 (levobupivacaine) and 13 (bupivacaine) while walking (VAS; 100 mm = worst pain imaginable) (ns). There was no difference in the use of peroral postoperative analgesics between the two groups. We conclude that racemic bupivacaine and its S-enantiomer levobupivacaine had similar efficacy when used as local infiltration anaesthesia in inguinal herniorrhaphy. PMID- 10365011 TI - Bolus dose remifentanil for control of haemodynamic response to tracheal intubation during rapid sequence induction of anaesthesia. AB - The effect of three bolus doses of remifentanil on the pressor response to laryngoscopy and tracheal intubation during rapid sequence induction of anaesthesia was assessed in a randomized, double-blind, placebo-controlled study in four groups of 20 patients each. After preoxygenation, anaesthesia was induced with thiopental 5-7 mg kg-1 followed immediately by saline (placebo) or remifentanil 0.5, 1.0 or 1.25 micrograms kg-1 given as a bolus over 30 s. Cricoid pressure was applied just after loss of consciousness. Succinylcholine 1 mg kg-1 was given for neuromuscular block. Laryngoscopy and tracheal intubation were performed 1 min later. Arterial pressure and heart rate were recorded at intervals until 5 min after intubation. Remifentanil 0.5 microgram kg-1 was ineffective in controlling the increase in heart rate and arterial pressure after intubation but the 1.0 and 1.25 micrograms kg-1 doses were effective in controlling the response. The use of the 1.25 micrograms kg-1 dose was however, associated with a decrease in systolic arterial pressure to less than 90 mm Hg in seven of 20 patients. PMID- 10365012 TI - Comparison of four methods for assessing airway sealing pressure with the laryngeal mask airway in adult patients. AB - We have compared four tests for assessing airway sealing pressure with the laryngeal mask airway (LMA) to test the hypothesis that airway sealing pressure and inter-observer reliability differ between tests. We studied 80 paralysed, anaesthetized adult patients. Four different airway sealing pressure tests were performed in random order on each patient by two observers blinded to each other's measurements: test 1 involved detection of an audible noise; test 2 was detection of end-tidal carbon dioxide in the oral cavity; test 3 was observation of the aneroid manometer dial as the pressure increased to note the airway pressure at which the dial reached stability; and test 4 was detection of an audible noise by neck auscultation. Mean airway sealing pressure ranged from 19.5 to 21.3 cm H2O and intra-class correlation coefficient was 0.95-0.99. Inter observer reliability of all tests was classed as excellent. The manometric stability test had a higher mean airway sealing pressure (P < 0.0001) and better inter-observer reliability (P < 0.0001) compared with the three other tests. We conclude that for clinical purposes all four tests are excellent, but that the manometric stability test may be more appropriate for researchers comparing airway sealing pressures. PMID- 10365013 TI - Epidural haematoma after removal of an epidural catheter in a patient receiving high-dose enoxaparin. AB - A patient developed an epidural haematoma 6 days after removal of an epidural catheter resulting in paraplegia and death. Insertion and removal of the epidural catheter during anticoagulation with prophylactic unfractionated heparin and subsequent administration of high-dose enoxaparin (Clexane), which commenced 3 days after catheter removal, were implicated. PMID- 10365015 TI - Comparison of basic methods in clinical studies and in vitro tissue and cell culture studies reported in three anaesthesia journals. AB - Tissue and cell culture (in vitro) studies reported in the 1997 issues of the British Journal of Anaesthesia, Anesthesia and Analgesia, and Anesthesiology were compared with groups of clinical studies selected at random from the same issues. Comparisons were of some basic aspects of study design and reporting that might lead to bias. The aspects examined were sample size, randomization and reporting of exclusions and withdrawals. Two groups of 53 articles were compared: sample size was smaller in in vitro than in clinical studies (median 6 vs 19); randomization was reported in five in vitro studies and in 37 studies; and failures were reported in two in vitro studies and in 43 clinical studies. This hinders interpretation of reported tissue and cell culture studies. Where possible, tissue and cell culture studies should be conducted, reported and assessed for publication to standards equivalent to those for clinical studies. PMID- 10365014 TI - Liability of laryngeal mask airway devices to thermal damage from KTP and Nd:YAG lasers. AB - We have compared the liability of four laryngeal mask airway (LMA) devices (standard, flexible, intubating and reusable) and a tracheal tube to thermal damage from KTP and Nd:YAG lasers at two power densities used commonly in airway surgery: 570 W cm-2 and 1140 W cm-2. Eighty-five airway devices were tested: 24 standard LMA (silicone-based), 12 flexible LMA (silicone-based, metal wires), 24 disposable LMA (PVC-based), one intubating LMA (silicone and steel-based) and 24 PVC-based tracheal tubes. Comparisons were made during laser strike to eight different targets: the unmarked and marked part of the airway device tube; the unmarked part of the airway device tube after application of blood; the cuff filled with air or methylene blue dye; the unmarked flexible LMA tube on or between the metal wires; and the epiglottic elevator bar of the intubating LMA. The laser strike was continued for 30 s and each target was tested three times. Three different, but identical, impact sites were used for each target. There was no ignition of any airway device with either power density or laser type. The silicone-based LMA were generally more resistant to flaring and penetration than the PVC-based LMA and tracheal tube, but the intubating LMA tube flared more rapidly with the KTP laser, and the disposable LMA cuff was more resistant to penetration. Print markings, blood and the metal wires of the flexible LMA reduced the thermal resistance of the tube. Filling the cuff with methylene blue dye increased the thermal resistance of all airway devices. We conclude that the silicone-based LMA devices were more thermal resistant to KTP and Nd:YAG laser strike than PVC-based devices with the exception of the disposable LMA cuff and the intubating LMA tube. PMID- 10365016 TI - Malignant hyperpyrexic syndrome. PMID- 10365017 TI - Early clinical experience with a newly formulated hydroxyethyl starch--Hextend. PMID- 10365018 TI - Pre-emptive analgesia--why the evidence is conflicting. PMID- 10365019 TI - Cisatracurium in a patient with atypical plasma cholinesterase. PMID- 10365020 TI - Epidural anaesthesia for caesarean section in an achondroplastic dwarf. PMID- 10365021 TI - Inadvertent inhalation anaesthesia during surgery under retrobulbar eye block. PMID- 10365022 TI - Reduce dose of NSAIDs in the elderly. PMID- 10365024 TI - Epidural analgesia during labour. PMID- 10365023 TI - Epidural analgesia during labour. PMID- 10365025 TI - Epidural bupivacaine and morphine on stump sensation in lower limb amputees. PMID- 10365026 TI - Postoperative cognitive deficit in the elderly surgical patient. PMID- 10365027 TI - Diabetic tractional papillopathy: a new (and true) nosological entity? PMID- 10365028 TI - Paediatric pseudophakia--choosing the implant power. PMID- 10365029 TI - Vitreopapillary traction in proliferative diabetic vitreoretinopathy [ssee comments]. AB - AIM: To present the clinical profile of a new entity in advanced proliferative diabetic vitreoretinopathy (PDVR). Mechanisms of vision loss due to vitreopapillary traction on the nasal optic disc are described, followed by an introduction of methods for prevention and treatment in such cases. METHODS: 17 patients with PDVR and traction on the nasal side of the optic disc, pallor of the optic nerve head, and reduced visual acuity were included in the study. Six patients were observed retrospectively and 11 patients prospectively before and after pars plana vitrectomy. Pre- and postoperative examinations included visual acuity, Goldmann's visual field, fluorescein angiography, and measurements of visual evoked potentials (VEP). RESULTS: During a postoperative follow up period of 3 to 24.5 months (mean 14.5 months) an improvement in optic disc appearance combined with an increased visual acuity (mean increase in VA = 0.171) was observed in 15/17 (88.3%) patients. In addition, 8/17 (47%) of these patients showed higher VEP amplitudes (mean 3.83 microV), and eight (6/8 of the same patients as VEP amplitudes) patients showed a reduction of latency (mean reduction 22.25 ms) during VEP assessment. CONCLUSION: These results suggest that vitreopapillary traction may damage the anterior optic nerve, via decreased axoplasmatic flow in the optic nerve fibres and/or mechanical reduction of perfusion in the posterior ciliary arteries. The effects of each mechanism appear to be reversible, but in the long term might lead to irreversible optic nerve atrophy. Therefore, in patients with vitreopapillary traction, early vitrectomy should be considered as a method to prevent optic neuropathy. PMID- 10365030 TI - Intraocular lenses in children: changes in axial length, corneal curvature, and refraction. AB - AIM: To assess changes in axial length, corneal curvature, and refraction in paediatric pseudophakia. METHODS: 35 eyes of 24 patients with congenital or developmental lens opacities underwent extracapsular cataract extraction and posterior chamber intraocular lens implantation. Serial measurements were made of axial length, corneal curvature, objective refraction, and visual acuity. RESULTS: For patients with congenital cataracts (onset < 1 year age) the mean age at surgery was 24 weeks. Over the mean follow up period of 2.7 years, the mean increase in axial length of 3.41 mm was not significantly different from the value of an expected mean growth of 3.44 mm (paired t test, p = 0.97) after correction for gestational age. In the developmental cataract group (onset > 1 year of age) the mean age at surgery was 6.4 years with a mean follow up of 2.86 years. This group showed a mean growth in axial length of 0.36 mm that was not significantly different from an expected value of 0.47 mm (paired t test, p = 0.63). The mean preoperative keratometry was 47.78 D in the congenital group and 44.35 D in the developmental group. At final follow up the mean keratometry in the congenital group was 46.15 D and in the developmental group it was 43.63 D. In eyes followed for at least 2 years, there was an observed myopic shift by 24 months postoperatively of 3.26 D in the congenital cases (n = 10) and 0.96 D in the developmental cases (n = 18). CONCLUSION: The pattern of axial elongation and corneal flattening was similar in the congenital and developmental groups to that observed in normal eyes. No significant retardation or acceleration of axial growth was found in the eyes implanted with IOLs compared with normal eyes. A myopic shift was seen particularly in eyes operated on at 4-8 weeks of age and it is recommended that these eyes are made 6 D hypermetropic initially with the residual refractive error being corrected with spectacles. PMID- 10365031 TI - Presumed ocular bartonellosis. AB - BACKGROUND: The spectrum of diseases caused by Bartonella henselae continues to expand and ocular involvement during this infection is being diagnosed with increasing frequency. METHODS: The clinical features and visual prognosis for 13 patients with intraocular inflammatory disease and laboratory evidence of bartonellosis were investigated. There were nine patients with neuroretinitis and four with panuveitis with positive antibody titres against B henselae determined by an enzyme immunoassay (IgG exceeding 1:900 and/or IgM exceeding 1:250). RESULTS: Positive IgG levels were found for eight patients and positive IgM levels for five. Despite animal exposure of 10 patients, only two (IgG positive) cases had systemic symptoms consistent with the diagnosis of cat scratch disease. Pathological fluorescein leakage of the optic disc was observed in all affected eyes. At 6 months' follow up, 3/18 (17%) affected eyes had a visual acuity of less than 20/100, owing to optic disc atrophy and cystoid macular oedema. 12 patients (17 eyes) were treated with antibiotics; visual acuity improved two or more Snellen lines for 9/17 (53%) eyes. CONCLUSIONS: The possibility of B henselae infection should be considered in patients with neuroretinitis and panuveitis (especially in cases with associated optic nerve involvement) even in the absence of systemic symptoms typical for cat scratch disease. PMID- 10365032 TI - Reproducibility of fundus autofluorescence measurements obtained using a confocal scanning laser ophthalmoscope. AB - AIM: To evaluate the reproducibility of the background fundus autofluorescence measurements obtained using a confocal scanning laser ophthalmoscope. METHODS: 10 normal volunteers and 10 patients with retinal disease were included in the study. One eye per subject was chosen randomly. Five images of the same eye of each individual were obtained, after pupillary dilatation, by two investigators using a confocal scanning laser ophthalmoscope. Background fundus autofluorescence was measured at 7 degrees temporal to the fovea in normal volunteers and between 7 and 15 degrees temporal to the fovea in patients. Within session reproducibility of the measurements obtained by each investigator and interobserver reproducibility were evaluated. RESULTS: For investigator 1 the median values of fundus autofluorescence obtained were 31.9 units for normal volunteers and 27.3 units for patients. The median largest differences in readings in normal volunteers was 5.7 units (range 1.4-13.5 units) and in patients 4.2 units (1.5-15.1 units). For investigator 2 the median values of fundus autofluorescence obtained were 28.9 units for normal volunteers and 27.4 units for patients. The median largest difference in readings in normal volunteers was 3.6 units (2.7-11.7 units), and in patients 4.1 units (1.5-9.3 units). The median interobserver difference in readings in normal volunteers was 3.3 units and for patients 6.6 units. The median greatest interobserver difference in measurements obtained for normal volunteers was 8.8 units (8.4-23.0 units) and for patients 11.1 units (7.1-40.8 units). CONCLUSION: Within session reproducibility of the measurements of background fundus autofluorescence was satisfactory. Although interobserver reproducibility was moderate, the variability of the measurements of fundus autofluorescence between observers appears to be small when compared with variation in fundus autofluorescence with age and disease. PMID- 10365033 TI - Age over 46 years does not affect the pressure lowering effect of trabeculectomy in primary open angle glaucoma. AB - BACKGROUND/AIMS: Previous reports have suggested that the success rate for trabeculectomy is poorer in younger age groups but these studies often have heterogeneous groups representing different types of glaucoma with variable surgical prognosis. Therefore, the relation between age and the success of trabeculectomy in the single diagnostic category of primary open angle glaucoma (POAG) without identifiable risk factors was examined for failure in the age range 46-85 years. METHODS: The records of 208 patients who had undergone a first trabeculectomy for POAG were examined retrospectively. Age ranged from 46 to 85 (mean 66.7 years). The outcome of surgery was examined at final available follow up and at 1 and 2 years after surgery. Trabeculectomy was considered a success if intraocular pressure was < or = 21 mm Hg with or without additional medical treatment ("cumulative" success) and an "absolute" success if intraocular pressure was < = 21 mm Hg without additional medical treatment. RESULTS: Cumulative success for trabeculectomy was 92.3% at final follow up and 96.6% at 2 year follow up; absolute success rate was 66.3% at final follow up and 71.6% at 2 years. There was no significant trend for greater success of trabeculectomy in the older age groups (cumulative success at 2 year follow up, chi 2 for linear trend 1.07 (p = 0.3) nor was the drop in intraocular pressure following surgery significantly greater with increasing age (analysis of variance for intraocular pressure lowering from presentation to 2 years' follow up (Kruskal-Wallis, t = 5.9, p = 0.55). Patients with pseudoexfoliation were excluded from the main analysis as these patients have been shown to have a lower final intraocular pressure following trabeculectomy, a finding which was confirmed in this study. PMID- 10365034 TI - Follow up of focal narrowing of retinal arterioles in glaucoma. AB - AIM: To evaluate whether focal narrowing of retinal arterioles increases with progressive glaucomatous optic neuropathy. METHODS: Focal narrowing of retinal arterioles and area of neuroretinal rim were morphometrically evaluated on colour stereo optic disc photographs of 59 patients with primary open angle glaucoma, 22 patients with normal pressure glaucoma, 11 patients with secondary open angle glaucoma, and 31 patients with ocular hypertension. Minimum follow up was 8 months. Focal arteriolar narrowing was quantified by calculating the ratio of the vessel width in the broadest to the narrowest vessel part. RESULTS: In the subgroup of patients with progressive glaucomatous optic nerve damage (n = 37), focal narrowing of retinal arterioles increased significantly (p < 0.005) with decreasing neuroretinal rim area. In the subgroup of patients with stable appearance of the optic disc (n = 86), focal narrowing of retinal arterioles did not change significantly (p = 0.79). The positive correlation between increasing focal thinning of retinal arterioles and progression of glaucomatous optic neuropathy was present, although not statistically significant, in all the glaucoma subtypes examined. The location of focal thinning of retinal arterioles did not change in the follow up. CONCLUSIONS: Focal narrowing of retinal arterioles increases significantly with progressive glaucomatous optic neuropathy, independent of the type of glaucoma. It is stable in patients with non-progressive glaucoma. The findings agree with previous reports on a higher degree of focal arteriole narrowing in eyes with pronounced optic nerve damage in comparison with those with moderate optic nerve atrophy or normal eyes. In the clinical management of patients with glaucoma, in some eyes, increasing focal arteriole narrowing may suggest progression of disease. PMID- 10365035 TI - Detection of optic disc change with the Heidelberg retina tomograph before confirmed visual field change in ocular hypertensives converting to early glaucoma. AB - AIM: To determine whether analysis of sequential optic disc images obtained with the Heidelberg retina tomograph (HRT) is able to demonstrate optic disc change before the development of reproducible field defects in a group of ocular hypertensive (OHT) patients converting to early glaucoma. METHODS: Two groups were analysed: (1) 13 eyes of 13 OHT patients who subsequently developed reproducible field defects (converters); and (2) 13 eyes of 11 normal control subjects. Two sequential optic disc images were obtained using the HRT (median separation between images was 12 months for the converters and 13 months for the normals). The second image in the converter group was obtained before confirmed visual field loss. The optic disc variables were analysed both globally and segmentally using HRT software version 1.11. The Wilcoxon signed rank test was used to determine if there were any significant differences between the variables of the two image sets. RESULTS: Significant optic disc change was demonstrated in the group of converters: (1) global variables: the cup area increased by 9.7%, the C/D area ratio increased by 10.5%, and the rim area decreased by 6.9%; (2) segmental variables: the superonasal cup area increased by 11.0%, the superonasal C/D area ratio increased 11.7%, and the inferonasal cup volume increased by 28.4%. The temporal rim volume decreased by 15.6%, the inferotemporal rim volume decreased by 23.6%, and the rim area in the superonasal and superotemporal segments decreased by 6.6% and 6.9% respectively. CONCLUSION: Analysis of sequential optic disc images on the HRT allowed the detection of glaucomatous change before confirmed visual field change in a group of OHT patients converting to early glaucoma. PMID- 10365036 TI - Comparison between laser scanning tomography and computerised image analysis of the optic disc. AB - AIMS: To study the interchangeability of the measurements of the optic disc topography obtained by one computerised image analyser and one confocal laser tomographic scanner. METHODS: One eye of 28 patients with glaucoma or glaucoma suspects was studied. All cases had simultaneous stereoscopic disc photographs taken with the fundus camera Topcon TRC-SS and optic disc examination with the Heidelberg retina tomograph (HRT) during the same visit. The optic disc photographs were digitised and analysed with the Topcon ImageNet (TI) system. Three variables of the optic disc topography provided by the TI and the HRT were compared--cup volume (CV), rim area (RA), and cup area to disc area ratio (CA/DA). RESULTS: The mean values of CV and RA provided by the TI (0.52 (SD 0.32) mm3 and 1.58 (0.39) mm2, respectively) were greater (p < 0.01) than the mean values of CV and RA determined by the HRT (0.32 (0.25) mm3, and 1.33 (0.47) mm2, respectively). The mean value of CA/DA provided by the TI (0.42 (0.14)) and the HRT (0.42 (0.18)) was similar (p = 0.93). Correlation coefficients between measurements obtained by the two methods ranged from 0.53 to 0.73. CONCLUSION: There was a significant discrepancy in the measurements of rim area and cup volume of the optic disc obtained by a computerised image analyser and a laser scanning tomograph. PMID- 10365037 TI - Preperimetric glaucoma diagnosis by confocal scanning laser tomography of the optic disc. AB - AIM: To evaluate the ability of confocal scanning laser tomography of the optic nerve head to detect glaucomatous optic nerve damage in ocular hypertensive eyes without visual field defects. METHODS: The study included 50 normal subjects, 61 glaucoma patients with glaucomatous changes in the optic disc and visual field, and 102 "preperimetric" patients with increased intraocular pressure, normal visual fields, and glaucomatous appearance of the optic disc as evaluated on colour stereo optic disc photographs. For all individuals, confocal scanning laser tomographs of the optic nerve head were taken using the Heidelberg retina tomograph (HRT; software 2.01). RESULTS: Almost all investigated HRT variables varied significantly (p < 0.05) between the normal eyes and preperimetric glaucoma eyes with pronounced overlap between the two study groups. Corresponding to the overlap, sensitivity and specificity values were relatively low when HRT variables were taken to differentiate between normal and preperimetric glaucoma eyes. At a given specificity of 95% highest sensitivities were found for the variables "rim area in the superior disc sector" (24.8%), "nerve fibre layer thickness in the inferior disc sector" (26.5%), and "rim volume in the superior disc sector" (25.5%). A multivariate approach increased sensitivity to 42.2% at a given specificity of 95%. For the glaucoma group highest sensitivity values were reached by rim volume in the superior disc sector (73.8%) and rim area (72.1%); the multivariate approach reached 83.6%. CONCLUSIONS: Owing to pronounced overlapping between the groups, confocal scanning laser tomography of the optic nerve head has relatively low diagnostic power to differentiate between normal eyes and preperimetric glaucoma eyes. One of the reasons may be the biological interindividual variability of quantitative optic disc variables. PMID- 10365038 TI - Course of exfoliation and simplex glaucoma after primary trabeculectomy. AB - AIM: To study the course of exfoliation and simplex glaucoma with respect to intraocular pressure (IOP) regulation and visual field survival after primary trabeculectomy. METHODS: Postoperative IOP regulation and complications were analysed prospectively in 95 patients. Mean follow up was 46 months. Visual field survival was studied by high pass resolution perimetry (HRP) in a subsample of 28 patients. RESULTS: Medical treatment was reinstated in 42% of exfoliation and in 36% of simplex glaucoma. In these patients, mean medicine free survival time, last untreated IOP, and mean IOP at the end of follow up were similar for both glaucoma types. Among patients with controlled postoperative IOP without added medication, mean IOP at the end of follow up was significantly lower in exfoliation glaucoma. Visual field deterioration and the pattern of complications were similar for both glaucoma types. CONCLUSION: The effect of trabeculectomy on IOP regulation was good in both types of glaucoma, and somewhat better in exfoliation glaucoma. The magnitude of IOP lowering could not separate patients with continued visual field deterioration from those in whom visual fields remained stable. Visual field preservation was similar for both glaucoma types. PMID- 10365039 TI - "Cyclodiode": results of a standard protocol. AB - AIMS: To analyse the results of intraocular pressure (IOP) reduction in refractory glaucoma following diode laser cyclophotocoagulation with a repeatable standard protocol. METHODS: 58 eyes of 53 patients were followed for 6-37 months (mean 19 months) after initial cyclodiode treatment. RESULTS: Mean (SD) pretreatment IOP for the 58 eyes was 33.0 mm Hg (10.7) reducing at final index visit to 16.7 mm Hg (7.8) (p < 0.0001). The mean antiglaucoma medication score per eye was significantly reduced from 2.4 (0.8) to 1.4 (1.0) at last index visit (p < 0.0001) with 91% of patients able to stop oral acetazolamide. 45% of eyes required more than one treatment and the overall mean per eye was 1.6 (range 1 5). Of eyes with visual acuity 6/60 or better pretreatment, 12 (32%) lost more than two lines of Snellen acuity and two eyes with poorer acuity initially dropped to NPL. Poor visual outcome was associated with the presence of diabetic retinopathy. Hypotony (IOP < 5 mm Hg) was noted in two eyes at the last index visit although neither had specific signs of the same. No phthisis was seen. CONCLUSION: The simple treatment protocol, repeated if necessary, appears relatively safe and effective at lowering IOP in eyes with refractory glaucoma. PMID- 10365040 TI - Prognosis of primary ab externo surgery for primary congenital glaucoma. AB - BACKGROUND: The strategy of pressure reducing surgery in primary congenital glaucoma has changed over the last decade. Ab externo filtering procedures--for example, trabeculectomy or trabeculotomy combined with trabeculectomy, have now been accepted even as primary intervention. METHODS: The authors reviewed 61 eyes in 35 consecutive patients with primary congenital glaucoma, who underwent different types of initial ab externo surgery between 1988 and 1996 (median follow up 36 months) to determine the efficacy of different surgical techniques and the influence of various risk factors. RESULTS: Trabeculotomy was performed in 17 eyes (27.9%), trabeculotomy with trabeculectomy in 15 eyes (24.6%), and trabeculectomy in 29 eyes (47.5%). Regarding age, preoperative intraocular pressure, corneal diameter, ocular axial length, and incidence of corneal haze the subgroups were comparable. Success rates of trabeculotomy, trabeculectomy, and a combined procedure did not significantly differ when assessed by life table analysis. Patient age under 3 months (p = 0.014) and an ocular axial length of 24 mm or more (p = 0.016) proved to be major risk factors for primary ab externo surgery failure. A second operation was necessary in 20 of 61 eyes (32.8%) during follow up. CONCLUSION: Prognosis of primary ab externo glaucoma surgery in primary congenital glaucoma seems to be governed more by the individual course and severity of the disease than by modification of surgical techniques. PMID- 10365041 TI - The eye in epidermolysis bullosa. AB - AIMS: To describe the ophthalmic findings in a large cohort of epidermolysis bullosa (EB) patients managed in one large specialist centre. METHODS: A case note review of consecutive patients seen at Great Ormond Street Children's Hospital. Data on the dermatological disease, ophthalmic history, and examination were collected and coded onto a data sheet. RESULTS: 181 patients: 50 (28%) simplex EB; 15 (8%) junctional EB; 28 (15%) autosomal dominant dystrophic EB; 72 (40%) autosomal recessive dystrophic EB; nine patients (5%) with dystrophic EB whose inheritance could not be ascertained; and seven cases (4%) of EB that could not be classified. Ocular problems were found in 12% (n = 6) of simplex patients and 40% (n = 6) of those with junctional disease. One patient (of 28) in the autosomal dominant dystrophic group had ocular involvement and 51% (37/72) of patients in the autosomal recessive dystrophic group had ophthalmic complications: corneal (25/72), lid ectropions (3/72), lid blisters (5/72), and symblepharon (3/72). CONCLUSION: Ophthalmic complications are common in EB overall but the incidence varies widely with subtype. Ophthalmic complications are the most severe in the dystrophic recessive and junctional subtypes where there is a need for extra vigilance. The major treatment modality was use of ocular lubricants. PMID- 10365042 TI - A new surgical technique for deep stromal, anterior lamellar keratoplasty. AB - AIMS: To describe a new surgical technique for deep stromal anterior lamellar keratoplasty. METHODS: In eye bank eyes and sighted human eyes, aqueous was exchanged by air, to visualise the posterior corneal surface--that is, the "air to endothelium" interface. Through a 5.0 mm scleral incision, a deep stromal pocket was created across the cornea, using the air to endothelium interface as a reference plane for dissection depth. The pocket was filled with viscoelastic, and an anterior corneal lamella was excised. A full thickness donor button was sutured into the recipient bed after stripping its Descemet's membrane. RESULTS: In 25 consecutive human eye bank eyes, a 12% microperforation rate was found. Corneal dissection depth averaged 95.4% (SD 2.7%). Six patient eyes had uneventful surgeries; in a seventh eye, perforation of the lamellar bed occurred. All transplants cleared. Central pachymetry ranged from 0.62 to 0.73 mm. CONCLUSION: With this technique a deep stromal anterior lamellar keratoplasty can be performed with the donor to recipient interface just anterior to the posterior corneal surface. The technique has the advantage that the dissection can be completed in the event of inadvertent microperforation, or that the procedure can be aborted to perform a planned penetrating keratoplasty. PMID- 10365043 TI - Iris cysts in children: classification, incidence, and management. The 1998 Torrence A Makley Jr Lecture. AB - BACKGROUND: Iris cysts in children are uncommon and there is relatively little information on their classification, incidence, and management. METHODS: The records of all children under age 20 years who were diagnosed with iris cyst were reviewed and the types and incidence of iris cysts of childhood determined. Based on these observations recommendations were made regarding management of iris cysts in children. RESULTS: Of 57 iris cysts in children, 53 were primary and four were secondary. There were 44 primary cysts of the iris pigment epithelium, 34 of which were of the peripheral or iridociliary type, accounting for 59% of all childhood iris cysts. It was most commonly diagnosed in the teenage years, more common in girls (68%), was not recognised in infancy, remained stationary or regressed, and required no treatment. The five mid-zonal pigment epithelial cysts were diagnosed at a mean age of 14 years, were more common in boys (83%), remained stationary, and required no treatment. The pupillary type of pigment epithelial cyst was generally recognised in infancy and, despite involvement of the pupillary aperture, also required no treatment. There were nine cases of primary iris stromal cysts, accounting for 16% of all childhood iris cysts. This cyst was usually diagnosed in infancy, was generally progressive, and required treatment in eight of the nine cases, usually by aspiration and cryotherapy or surgical resection. Among the secondary iris cysts, two were post-traumatic epithelial ingrowth cysts and two were tumour induced cysts, one arising from an intraocular lacrimal gland choristoma and one adjacent to a peripheral iris naevus. CONCLUSIONS: Most iris cysts of childhood are primary pigment epithelial cysts and require no treatment. However, the iris stromal cyst, usually recognised in infancy, is generally an aggressive lesion that requires treatment by aspiration or surgical excision. PMID- 10365044 TI - AIDS related eye disease in Burundi, Africa. AB - AIMS: To determine the prevalence of ocular manifestations in AIDS patients hospitalised in Bujumbura, Burundi, according to their CD4+ lymphocyte count, serological status for CMV and VZV, and general health status. METHODS: Prospective study of 154 consecutive patients who underwent general and ophthalmological examinations, including dilated fundus examination. AIDS was diagnosed on the basis of Bangui criteria and HIV-1 seropositivity. CD4+ lymphocyte counts were determined by the Capcellia method. CMV and VZV antibodies were detected with ELISA methods. RESULTS: The mean age was 37 (SD 9) years and 65% of the patients were male. Active tuberculosis was the most frequent underlying disease (61%). Almost all the patients (99%) were seropositive for CMV and VZV. Among the 115 patients for whom CD4+ lymphocyte counts were available, 86 (75%) had more than 100 cells x 10(6)/l. Ocular involvement comprised 16 cases of microangiopathy, six of opalescence of the anterior chamber, five of retinal perivasculitis, two of zoster ophthalmicus, two of viral retinitis, and one of opalescence of the vitreous. CONCLUSION: In Africa, the prevalence of ocular involvement in HIV infection is far lower than in Europe and the United States, possibly because most African patients die before ocular opportunistic infections occur. PMID- 10365045 TI - Evaluation of visual outcome of cataract surgery in an Indian eye camp. AB - AIM: To evaluate the results of cataract surgery performed in a rural Indian eye camp. METHOD: The pre- and postoperative visual acuities and surgical complications were recorded prospectively in 6383 eyes undergoing cataract extraction for age related cataract in rural eye camps held in northern India in 1993-4. The best visual acuity and cause of poor outcome were recorded on 3908 eyes seen at 6 weeks' follow up. RESULTS: Of 6383 operated eyes 94.8% had a visual acuity of less than 3/60 preoperatively, and 41% of the procedures were performed on patients who were bilaterally blind (less than 3/60 better eye). At discharge with standard aphakic spherical spectacles, 11.3% of eyes had an acuity of less than 6/60 (poor outcome), and 25.9% had an acuity of 6/18 or better. At 6 weeks' follow up 3908 eyes were examined (61.2%), of which, with best correction, 4.3% had poor outcome (acuity of less than 6/60) and 79.9% obtained 6/18 or better. Pre-existing eye pathology was responsible for poor outcome in 3.0% of eyes and surgical complications in 1.3% of eyes, of which corneal decompensation was the major cause (0.5%). In 237 eyes which received an intraocular lens implantation (IOL) in the camp, the visual acuity at discharge was 6/18 or better in 44.5% of eyes improving to 87.9% in the 157 eyes which were seen at 6 weeks' follow up. Poor outcome (less than 6/60) was seen in 5.7% of the eyes with an IOL at discharge improving to 1.9% at follow up. CONCLUSION: This evaluation suggests that it is possible to obtain acceptable results from cataract extraction with experienced ophthalmologists in well conducted Indian eye camps. Better correction of aphakia at discharge from the camp would improve the immediate visual results, which is important as a significant number of patients do not return for follow up. The use of posterior chamber IOLs in the eye camp by experienced ophthalmologists, appeared to give satisfactory results, although further evaluation with a larger series of cases and more surgeons is required before it can be recommended. PMID- 10365046 TI - Topographic anatomy of the eyelids, and the effects of sex and age. AB - AIMS: To describe the effects of sex and age on eyeball, eyelid, and eyebrow position. METHODS: A cross sectional cohort study was performed in which both eyes of 320 normal subjects aged between 10 and 89 years were included. Of each 10 year age cohort, there were 20 men and 20 women. Frontal, as well as lateral, slides were taken of both eyes. On projected slides, a reference line through the medial canthi and vertical lines through the pupil centre and the lateral canthus were constructed. Using these lines, we measured the size of the horizontal eyelid fissure, the distance from the reference line to the pupil centre and to the lateral canthus, the distance between the pupil centre and the upper and lower eyelid margin, and the distance between the upper eyelid margin and the skin fold and eyebrow. On lateral slides, the distance between the lateral canthus and the anterior corneal surface was measured. RESULTS: Between the ages of approximately 12 and 25 years, the horizontal eyelid fissure lengthened 3 mm, while the position of other eyelid structures remained virtually unchanged. Between the average ages of 35 and 85 years, the horizontal eyelid fissure gradually shortened again by about 2.5 mm. Meanwhile, the distance between the lateral canthal angle and the anterior corneal surface decreased almost 1.5 mm. Aging caused an increase of the distance between the pupil centre and the lower eyelid of about 1 mm in men, and 0.5 mm in women. Aging also caused a higher skin crease and raised eyebrows in men and women, but it did not affect the position of the pupil centre and the lateral canthus. Men showed an 0.7 mm larger horizontal eyelid fissure than women. In women, however, the eyebrows were situated about 2.5 mm higher than in men. CONCLUSION: Aging mainly affects the size of the horizontal eyelid fissure, which lengthens by about 10% between the ages of 12 and 25, and shortens by almost the same amount between middle age and old age. Aging causes sagging of the lower eyelid, especially in men, and a higher skin fold and eyebrow position in both sexes. Aging does not affect the position of the eyeball proper, or of the lateral canthus. PMID- 10365047 TI - Measurement of retinal nerve fibre layer by scanning laser polarimetry and high pass resolution perimetry in normal tension glaucoma with relatively high or low intraocular pressure. AB - AIMS: To determine whether any differences may exist in the relation between the neural capacity as determined by high pass resolution perimetry and the thickness of the retinal nerve fibre layer (RNFL) in patients having normal tension glaucoma (NTG) with a relatively high intraocular pressure (IOP) between 16 and 21 mm Hg (HNTG) v those with a lower IOP below 15 mm Hg (LNTG). METHODS: Scanning laser polarimetry and high pass resolution perimetry were performed in 20 eyes of 20 patients with HNTG and 21 eyes of 21 patients with LNTG. The correlation between total and regional thickness of the peripapillary RNFL and the corresponding total and regional neural capacity with linear regression analysis were evaluated. RESULTS: Overall, although the total RNFL thickness was not significantly correlated with the total neural capacity, the RNFL thickness in each of the superior and inferior quadrants was significantly correlated with the corresponding regional neural capacity (r = 0.44, p = 0.0045; r = 0.39, p = 0.0126 for each). The RNFL thickness in each of the superior and inferior quadrants in the HNTG group was significantly correlated with the corresponding regional neural capacity (r = 0.52, p = 0.0196; r = 0.49, p = 0.0286 for each). No significant correlation between neural capacity and the RNFL thickness was observed either globally or regionally in the LNTG group. CONCLUSION: The degree of the correlation between neural capacity as determined by high pass resolution perimetry and thickness of the RNFL as measured by scanning laser polarimetry appeared to differ in NTG patients with an IOP higher than 15 mm Hg v those with a lower IOP. PMID- 10365050 TI - Viridans group Streptococcus subretinal abscess. PMID- 10365048 TI - Early drusen formation in the normal and aging eye and their relation to age related maculopathy: a clinicopathological study. AB - AIM: To describe the early formation of drusen and their relation to normal aging changes at the macula and to the development of age related maculopathy (ARM). METHOD: Histopathological features of 353 eyes without histological evidence of ARM are described and correlated with the clinical appearance. In addition, 45 of these eyes were examined by transmission electron microscopy. RESULTS: Drusen were detected histopathologically in 177 (50%) eyes but were seen clinically in only 34% of these. Drusen were mainly small hard drusen with an occasional soft distinct drusen: no soft indistinct drusen were seen. Only those drusen deposits larger than 25-30 microns in diameter were detectable clinically. Preclinical drusen in eyes with only an occasional drusen were seen on electron microscopy as entrapment sites of coated membrane bound bodies which formed adjacent to the inner collagenous zone of Bruch's membrane. In contrast, preclinical drusen deposits in eyes with many drusen were seen as accumulations of amorphous material which appeared hyalinised by light microscopy. A distinct feature were rows of dense hyalinised microdrusen (1-2 microns in diameter), over which larger globular hyalinised drusen formed. CONCLUSION: Histological and ultrastructural examination can recognise and distinguish the earliest drusen formed as a result of normal aging from those associated with ARM. In eyes without diffuse deposits, histologically all drusen were of the hard hyalinised variety or their derivatives; no soft drusen composed of membranous debris were found. These findings support and explain those of other authors who do not consider the presence of a few small hard drusen to be a risk factor for the development of ARM. PMID- 10365051 TI - Inferior oblique myectomy using monopolar cutting diathermy resulting in bilateral retinal scarring. PMID- 10365049 TI - Should diabetic patients be screened for glaucoma? DARTS/MEMO Collaboration. PMID- 10365052 TI - Ultrasound biomicroscopy in juvenile xanthogranuloma of the iris. PMID- 10365053 TI - A traumatic "peripheral iridotomy" protects against pigment dispersion and glaucoma. PMID- 10365054 TI - Inadvertent exposure of corneal endothelium to 5-fluorouracil. PMID- 10365055 TI - Red-free light in applanation tonometry. PMID- 10365056 TI - Injury to the globe during periocular anaesthesia. PMID- 10365057 TI - Polymerase chain reaction in the diagnosis of bacterial endophthalmitis. PMID- 10365058 TI - Effect of amblyopia on employment prospects. PMID- 10365059 TI - Patients leaving hospital after administration of radioactive substances. Working Party of the Radiation Protection Committee of the British Institute of Radiology. PMID- 10365060 TI - Vascular ultrasound: yet another casualty? PMID- 10365061 TI - Forensic radiology. AB - Imaging techniques are a powerful tool in forensic science. Medical examiners and forensic anthropologists are less versed in the finer points of radiology than radiologists; nevertheless they are required to interpret findings from imaging studies to further medico-legal investigations. The forensic investigator often should call upon the radiologist whose expertise might prove invaluable in forensic consultations. The radiologist should be aware of the importance of storing radiographs over prolonged periods of time and of efficient record keeping methods, because various legal problems may require the radiographs for additional interpretation or for their presentation in court. Some of the main issues that might be encountered in forensic radiology are discussed in this review. PMID- 10365062 TI - The value of serial Doppler ultrasound as a predictor of clinical outcome and the need for transplantation in fulminant and severe acute liver failure. AB - The aim of this study was to document the changes in Doppler ultrasound variables of the hepatic artery and portal vein in fulminant and severe acute liver failure, and to assess their prognostic significance. 18 adult patients with fulminant and severe acute liver failure underwent serial Doppler sonography, in the early stages after presentation. 12 hourly measurements of hepatic artery resistance index (HARI), spleen length, portal vein cross-sectional area, time average velocity (TAV) and flow volume were performed. Mean HARI (p = 0.03) and mean maximum HARI (p = 0.03) were significantly higher in those who fulfilled criteria for liver transplantation. Increased portal vein flow was demonstrated, although the difference between the groups was not significant. A significant increase in portal vein cross-sectional area (p < 0.02) and spleen length (p < 0.02) was demonstrated. In summary, an increase in portal blood flow to the damaged liver has been demonstrated. The mean HARI is significantly higher in patients who fulfil transplant criteria and may possibly be used as an indicator of poorer prognosis and the need for liver transplantation in acute severe and fulminant liver failure. PMID- 10365063 TI - Ultrasound, CT and colonoscopy of colonic cancer. AB - Barium enema and colonoscopy are commonly used for the investigation of suspected colonic cancer. These techniques are relatively invasive and both the investigation and the preceding bowel preparation are demanding, particularly in the elderly. A prospective, blinded trial was conducted to compare ultrasound (US) and CT with colonoscopy. CT and colonoscopy were performed on 50 patients with symptoms suggesting colonic cancer. Both radiological investigations were performed prior to the bowel preparation for colonoscopy. US was performed without any preparation and oral contrast medium was the only preparation used for CT. Colonoscopy detected six cancers, all of which were diagnosed by both US and CT. In addition, US and CT diagnosed a further cancer not seen on colonoscopy due to an incomplete study. US had a sensitivity and specificity of 100% and CT a sensitivity of 100% and a specificity of 84%. US and CT were poor at diagnosing polyps. If the detection of polyps greater than 2 cm is included then US sensitivity falls to 67% and CT sensitivity falls to 89% and specificity rises to 88%. In conclusion, both US and CT are possible alternatives to colonoscopy in the investigation of symptomatic patients with suspected colonic cancer. The use of these techniques could markedly reduce the need for colonoscopy in this patient population with attendant cost savings. Non-invasive imaging has particular advantages in the elderly who cope poorly with both the bowel preparation and the procedure. PMID- 10365064 TI - Biliary imaging by spiral CT cholangiography--a retrospective analysis. AB - CT cholangiography employs spiral CT scanning after the administration of biliary contrast medium to generate three-dimensional images of the biliary tract. A retrospective review of 61 patients who had undergone CT cholangiography was performed to determine the technical efficacy and the clinical utility of the technique. The results of CT cholangiography were analysed retrospectively to determine technical success rate of the imaging procedure and to correlate imaging diagnosis with results of other diagnostic procedures and with clinical follow-up. 60 of the 61 examinations produced technically satisfactory images. 21 of these examinations showed bile duct abnormalities. (12 stones, 6 duct dilatation to papillary level, 2 post-operative strictures and 1 case of sclerosing cholangitis). The remaining 39 cases showed no bile duct abnormality. In 59 of the 60 cases, subsequent investigations and follow-up supported the CT cholangiographic diagnosis. CT cholangiography is a robust technique for imaging of the biliary tract and is a valuable addition to the battery of non-invasive biliary investigations. PMID- 10365065 TI - CT derived Patlak images of the human kidney. AB - This study aimed to produce Patlak images of the kidney from dynamic CT data and to determine whether such images are substantially affected by fluid movement between renal tubular segments. Renal permeability was measured in 31 kidneys by applying Patlak analysis to time-density data from kidney and aorta during dynamic CT. Permeability parameters were correlated against plasma urea. The renal region (cortex or medulla) with the greatest permeability was determined from parametric images generated using pixel by pixel analysis. The mean value for whole kidney permeability was 517.5 microliters min-1 ml-1. A correlation was found between whole kidney permeability and plasma urea (p < 0.01). Permeability values were highest in the renal medulla in 24 (77%) kidneys. The higher medullary values of permeability are artefactual, resulting from movement of fluid and contrast medium between cortex and medulla. Although Patlak images do not reflect true intrarenal permeability values, the apparent medullary permeability may provide diagnostically useful information about the concentrating ability of the kidney. CT measurements of whole kidney permeability reflect filtration function but the apparent intrarenal variations in permeability will result in measurement errors dependent upon the relative amounts of renal cortex and medulla included in the CT slice studied. PMID- 10365066 TI - Embolization of uterine fibroids. AB - 14 patients have undergone uterine artery embolization of uterine fibroids. No complications related to the procedure have occurred. In all six of the cases with available data, there has been a reduction in fibroid volume as measured with MRI at 2 months post procedure (average reduction 43%). Technical aspects of this promising procedure are described. PMID- 10365067 TI - The use of soft and flexible guidewires in the treatment of chronic total coronary occlusions by activated guidewire angioplasty. AB - Activated guidewire angioplasty (AGA) is a new technique which has been designed to assist in angioplasty of total occlusions. The purpose of this study was to determine the safety and efficacy of using flexible relatively soft guidewires (floppy wires) in conjunction with this technique and also to determine the predictors of lesion crossing and final success by this technique in patients with chronic total coronary occlusions. 73 patients with 73 chronic total coronary occlusions in whom coronary angioplasty using conventional techniques had failed were treated with AGA using floppy guidewires. The success of crossing these lesions was 65.7% (48/73) resulting in a final angioplasty success of 56.1% (41/73). Angioplasty success was reduced compared with crossing success in seven arteries in which complications occurred during balloon angioplasty. Multiple stepwise logistic regression analysis identified the location of the occlusion (right coronary artery, p = 0.005) as independent predictor of crossing success of this technique and the male gender (p = 0.03), the duration of occlusion (p = 0.05), the lesion length (p = 0.01) and the location of the occlusion (right coronary artery, p = 0.02) as independent predictors of final procedural success of the method. PMID- 10365068 TI - 60 Gy isodose volumes in carcinoma of the cervix and their relation to the geometry of uterine tube and ovoids. AB - The International Commission on Radiation Units and Measurements recommends the use of 60 Gy isodose volumes for reporting doses in the intracavity treatment of carcinoma of the uterine cervix. This study was aimed at determining the variation in isodose volumes while using different sizes of intrauterine tubes and ovoids, with different applicator geometries. It was based on the treatment plans of 175 patients with cervical cancer, treated with low dose rate intracavitary brachytherapy with or without additional external beam radiotherapy. The volumes encompassed by the 60 Gy isodose curves were calculated using the Nucletron planning system. Applicator positions in 15 patients who were treated to the same point A dose, using 6 cm intrauterine tube and medium ovoids, were recorded. This was to discover how variations in applicator geometry influences isodose volumes. The 60 Gy isodose volumes increased with increasing point A dose. For a constant point A dose prescription, reference isodose volume increased with ovoid applicator size used, but showed no consistent variation with the length of intrauterine tube. There were individual variations in the isodose volumes within a standard set-up (same sized intrauterine tube and ovoids and same point A dose), due to variations in applicator geometry. Displacement of the ovoids changed the volumes encompassed by the reference isodose. There are significant variations in the volumes encompassed by the 60 Gy isodose during intracavitary treatment using a standard set-up, while treatment using applicators of different sizes can give equivalent values of 60 Gy isodose volume. 60 Gy isodose volumes may hence be useful in dosimetric comparisons but have a limited role in predicting clinical response. PMID- 10365069 TI - Patient doses from barium meal and barium enema examinations and potential for reduction through proper set-up of equipment. AB - Patient doses for barium meal and barium enema examinations, performed at two Greek hospitals, were measured using a dose-area product meter. The results were analysed to obtain the contributions of fluoroscopy and radiography to the dose as well as a number of other dose related parameters for each examination. The doses observed are within the range of values reported by other authors and comply with the dose reference levels (DRLs), proposed from relevant surveys in the UK and The Netherlands. However, comparison between the two hospitals revealed significant differences in the contributions to dose from the various parts of the examinations. To determine the reasons for these differences, measurements of dose related parameters were made using a Plexiglas phantom and standard clinical X-ray machine settings. Factors contributing to increased dose delivery were determined and recommendations have been made concerning ways in which doses might be reduced in each hospital, without degradation of the diagnostic quality of these examinations. PMID- 10365070 TI - Calculating shielding requirements in diagnostic X-ray departments. AB - Structural radiation protection for diagnostic X-ray facilities is most commonly performed following the recommendations of the National Council on Radiation Protection and Measurements Report No. 49. A number of analytical methods have already been developed to improve the design of these facilities. Specifically, these methods reassess shielding calculations in X-ray areas with respect to the methodology of the calculation of the barrier thickness and the number of sources considered in the area. Thus, they generate an overall solution for the cases met at the medical radiation structural design. This paper presents an extension of an existing method for calculating shielding requirements, for multiple X-ray tubes in a room operated at various beam qualities. The methodology computes the required shielding thickness such that the exposure behind it stays below a desired value. The presented method eliminates the overestimation of added shielding thickness which may occur using the other methods already mentioned. A user-friendly windows-based program has also been developed to assist shielding computations. PMID- 10365072 TI - Two-dimensional kinetic models for radiotherapy planning quality assurance. AB - For treatment planning, the visualization of the direction and orientation of a beam, specified by a set of machine angles, can sometimes be very difficult. A set of simple two-dimensional kinetic models has been prepared. These show the angular and field conventions of the local linear accelerators. PMID- 10365073 TI - The X-ray and electron benchmarking of the Monte Carlo codes MCNP-4A and 4B on different computers. AB - MCNP (Monte Carlo N-Particle) is a Monte Carlo transport code which has been of widespread use in modelling the dosimetry of ionizing radiations. The most recent version (4B) features improved electron transport compared with the previous version 4A. The processing time required by a number of computing systems to carry out X-ray and electron transport calculations using both versions of the code was compared. Version 4A was installed onto a Dec Alpha Server 8200, a personal computer (Pentium 90 MHz), and a Sun Sparc20, 10, 4 and 1+. MCNP-4B was also installed onto the Sun Sparc20. The benchmark tests consisted of determining the transmission of 2 MeV X-rays and 30 MeV electrons through lead. It was found that the Dec Alpha Server 8200 was the fastest computing platform, and the Sun Sparc1+ was the slowest for both tests. The difference in computational speed between different platforms was not matched by the corresponding differences in price. The time required by version 4B to complete the X-ray and electron benchmark tests was found to be 1.4 and 2.3 times greater than version 4A, respectively, without any difference in the results of the calculation for each type of radiation. This suggests that in cases where computing time is important, it may be preferable to use version 4A instead of 4B. PMID- 10365071 TI - Report of an image quality and dose audit according to directive 97/43/Euratom at Spanish private radiodiagnostics facilities. AB - An audit of Spanish private medicine radiodiagnostics facilities has been carried out, based partly on Spanish legislation relating to European Directives on health protection against ionizing radiation risks in medical exposure. The study included an appraisal of infrastructure and equipment, and aspects of quality assurance and radiation protection, by means of data collected through surveys. Of the 51 centres audited, a sample of 24 X-ray rooms was chosen, then an external evaluation with regard to image quality and patient dose was performed, by an advisory board of radiologists and medical physicists. The methodology used was similar to that of the group of European Union experts in European dose evaluation and image quality trials. Chest, abdomen, lumbar spine and breast examinations were monitored. Doses were measured with thermoluminescent dosimeters. A third of the X-ray rooms evaluated reached or exceeded dose reference values, and in a third of the cases the image quality left considerable room for improvement. Breast and chest examinations showed themselves to be the hardest to perform, not only as a result of exceeding the reference doses, but also due to failure to meet good image quality standards. PMID- 10365074 TI - Peritoneal abscess formation as a late complication of gallstones spilled during laparoscopic cholecystectomy. AB - The case is described of a 74-year-old woman who presented with an abdominal abscess 1.5 years after laparascopic cholecystectomy. CT and ultrasound showed the presence of gallstones within the abscess. Spillage of gallstones from perforation of the gallbladder is a well recognized complication of laparascopic cholecystectomy, although subsequent abscess formation is unusual especially after a long delay as in this case. PMID- 10365075 TI - Persistent hypophyseal (craniopharyngeal) canal. AB - Two cases with a broad persistent hypophyseal canal connecting pituitary fossa and nasopharynx are presented. Both had nasopharyngeal surgery in early childhood and presented later with hypopituitarism. Coronal CT demonstrated the defects with no visible pituitary tissue. The spectrum of basal skull defects is discussed. Children with midline nasal polyps should have CT or MRI of the skull base prior to surgery to prevent inadvertent hypophysectomy. PMID- 10365076 TI - Imaging findings in primary carcinoid tumour of the liver with gastrin production. AB - We present a 57-year-old man with Zollinger-Ellison syndrome who had undergone total gastrectomy 12 years previously. At that time, a cystic mass in the left lobe of the liver was palpated but was not removed. The patient currently had high serum gastrin levels. Abdominal ultrasound, CT and MR images showed a well defined liver mass with solid and cystic components. The lesion was resected and a primary hepatic carcinoid tumour was diagnosed. Post-operatively, serum gastrin levels were normal. A primary liver carcinoid may appear as multicystic liver mass with solid components. Careful exclusion of a primary tumour elsewhere is required to establish the diagnosis of this rare entity. PMID- 10365077 TI - Acute pan-costochondritis demonstrated by gallium scintigraphy. AB - Costochondral inflammation is a rare clinical finding. A case is presented, in which acute chostochondritis is demonstrated by gallium scinitigraphy. PMID- 10365078 TI - MR imaging of synovial tumours and tumour-like lesions. AB - MR is the imaging method of choice for detection, staging and follow-up of synovial tumours and tumour-like conditions. The majority of lesions have non specific features but careful analysis of lesion morphology and signal characteristics reduces the differential diagnosis and leads to a specific diagnosis in some cases. PMID- 10365079 TI - An unlucky break. PMID- 10365080 TI - Awareness by radiology staff of the difference in radiation risk from two opposing lateral lumbar spine examinations. PMID- 10365082 TI - Protective effect of pentoxifylline plus thalidomide against septic shock in mice. AB - Mortality caused by septic shock in experimental animals is reduced by thalidomide, an inhibitor of tumour necrosis factor alpha. Another drug that could act on the pathophysiological mechanisms of septic shock is pentoxifylline, an inhibitor of platelet aggregation that increases the flexibility of the erythrocyte membrane and has fibrinolytic activity. We studied the effect of pentoxifylline alone and combined with thalidomide in septic shock; 97 NIH mice were injected with lipopolysaccharides of Salmonella abortus equi and D galactosamine. Animals were separated in 4 groups; group A (n = 20) was used as control, group B (n = 15) received thalidomide 50 mg/kg, group C (n = 20) received pentoxifylline 40 mg/kg, and group D (n = 15) received thalidomide plus pentoxifylline. Mortality was recorded every hour. Additionally, 5 animals from each group were sacrificed 8 h after the induction of septic shock for histological analysis of heart, lung, brain, kidney, small intestine, adrenal glands and liver. Microscopic findings were rated as absent, mild, moderate and severe damage. In control animals histological analysis showed intense haemorrhage and necrosis in all organs studied. When compared with controls, treatment with pentoxifylline plus thalidomide reduced mortality (P < 0.03). The tissue damage was less severe in animals from the groups that received pentoxifylline or pentoxifylline plus thalidomide (P < 0.05). Pentoxifylline seems to potentiate the beneficial effects of thalidomide, reducing mortality and attenuating the pathological changes produced by septic shock. PMID- 10365081 TI - Influence of host microvascular environment on tumour vascular endothelium. AB - This review will focus on the tumour microvascular endothelium; how it is derived, modulated by angiogenic factors, and how the structure and function is influenced by the host tissue microenvironment. PMID- 10365083 TI - Degradation of Japanese encephalitis virus by neutrophils. AB - The ability of neutrophils to degrade the phagocytosed Japanese encephalitis (JE) virion, via triggering of the respiratory burst and generation of toxic radicals has been investigated. JEV or JEV-induced macrophage derived factor (MDF) induces increase in intracellular oxidative signals with generation of superoxide anion (O2-), via activation of cytosolic NADPH and subsequent formation of hydrogen peroxide, with maximum activity on day 7 post infection. The response was sensitive to anti-MDF antibody treatment. Further, the study revealed rapid degradation of phagocytosed JE viral protein and nucleic acid. The viral protein degradation was partially dependent on the generation of toxic oxygen species as it could be abrogated by pretreatment of the cells with staurosporine. PMID- 10365084 TI - Colchicine therapy for hepatic murine schistosomal fibrosis: image analysis and serological study. AB - Colchicine in a dose of 200 micrograms kg body weight/day (5 days/week) was administered to groups of Schistosoma mansoni infected mice 12 weeks post infection, either alone or following previous praziquantel therapy at the 8th week of infection. Certain groups received colchicine for 6 weeks and others received it for 10 weeks. Colchicine alone did not significantly change the light microscopic appearance of schistosomal liver fibrosis, or hepatic collagen content estimated histomorphometrically, and did not reduce the elevated IL-2 serum level. Colchicine induced hepatic injury consisted of intense inflammatory reaction in granuloma and portal tracts, hepatocytic degeneration, and elevation of serum AST and ALT levels. Colchicine seemed to postpone granulomatous reaction healing and collagen deposition rather than inhibiting collagen formation or degrading it. Colchicine inhibited proliferation of hepatocytes of infected mice by expanding G2-M phases of cell cycle, thus reduced Ag NOR count and raised cell ploidy and cyclic AMP serum level. Subsidence of schistosomal infection by praziquantel prior to colchicine therapy greatly reduced inflammatory cellular reaction, significantly diminished hepatic collagen deposition and serum IL-2 level, minimized the elevated nuclear ploidy and cyclic AMP serum level that followed colchicine therapy when administered alone. PMID- 10365085 TI - Colonization of the respiratory and genital tracts of female mice with Mycoplasma pneumoniae and protection afforded to the genital tract by prior respiratory colonization. AB - Mycoplasma pneumoniae, strain MY 12763, colonized the throats of 6 BALB/c female mice for at least 18 days after intranasal inoculation. The same strain colonized, in high titre, the vagina of 9 of 10 progesterone-treated BALB/c mice for at least 35 days, but none of 10 oestradiol-treated mice. Mice were less susceptible to genital-tract colonization with the multiple broth-passed FH strain of M. pneumoniae, only 3 of 10 becoming colonized. The 6 mice inoculated intranasally with strain MY 12763 were immune to genital-tract colonization with the same strain, whereas 10 mice without respiratory-tract colonization were susceptible. Protection of the genital tract in this way was at least as effective as that afforded by previous genital-tract colonization. In a further experiment, 26 immunocompetent BALB/c mice colonized previously in the respiratory tract were resistant to vaginal colonization, whereas 20 BALB/c nude mice, similarly colonized in the respiratory tract, were susceptible in the vagina, illustrating the importance of cell-mediated immunity. The possible relevance of the findings to the human situation is discussed. PMID- 10365086 TI - Decreased prostaglandin E2 synthesis by lung fibroblasts isolated from rats with bleomycin-induced lung fibrosis. AB - In order to clarify the mechanism of pulmonary fibrosis, we examined the functional changes of lung fibroblasts in bleomycin (BLM)-induced pulmonary fibrosis. Lung fibroblastic cells were obtained from rat lungs after an intratracheal treatment of BLM or saline. The spontaneous proliferation of BLM treated rat fibroblasts (BRF), which was estimated by 3H-TdR incorporation and direct cell counting, was significantly more rapid than that of normal saline treated rat fibroblasts (NRF). Next, we investigated prostaglandin (PG) E2 synthesis by BRF and NRF, with or without stimulation by interleukin (IL)-1 alpha, and found that PGE2 production by BRF was significantly less than that by NRF. There was no significant difference in cyclooxygenase (COX) activity and COX 2 mRNA level between BRF and NRF, indicating that the change in PGE2 production was independent of COX, a rate-limiting enzyme for the production of PGE2. These results suggest that the proliferation of fibroblasts is down-regulated by PGE2 released from themselves in normal lungs in an autocrine fashion, thus the decreased PGE2 production observed in lung fibroblasts from rats with BLM-induced pulmonary fibrosis may result in the excessive fibroblast proliferation in this disorder. Overall, these findings throw some light on the mechanism of development of BLM-induced pulmonary fibrosis. PMID- 10365087 TI - Low cytoplasmic pH causes fragmentation and dispersal of the Golgi apparatus in human hepatoma cells. AB - The centrosomal localization of the Golgi apparatus in interphase cells is thought to be maintained by retrograde microtubule-based motility. It is well established that, when intracellular pH is lowered, lysosomes and endosomes, also showing pericentrosomal localization, translocate towards the plus ends of microtubules within 15 min. In this study, we found that prolonged incubation in low pH medium (pH 6.6) with 20 mM Na acetate induced the fragmentation and dispersal of the Golgi apparatus in the human hepatoma cell line PLC/PRF/5. The fraction of Golgi-dispersed cells increased in a time-dependent manner, and reached over 60% after the 16-h incubation. The cytoplasmic pH was dropped to approximately 7.10. Replacement with normal pH medium restored the structure and localization of the apparatus within 30 min. In the low pH condition, the microtubular network and endoplasmic reticulum appeared normal, and cytoplasmic dynein was still bound to the fragmented Golgi membranes. These findings suggest that low cytoplasmic pH suppresses the retrograde movement of the Golgi apparatus as well as that of lysosomes and endosomes. PMID- 10365088 TI - The development of Lewisite vapour induced lesions in the domestic, white pig. AB - Studies performed in the past in our laboratory have detailed the development of sulphur mustard lesions in the domestic, white pig using small glass chambers to achieve saturated vapour exposure under occluded conditions. We have now used this experimental model to produce cutaneous lesions for detailed histopathological studies following challenge with lewisite. Histological examination of resulting lesions have revealed that although the overall pattern of lesion development is similar to that seen following mustard challenge, the time-course of cellular events is very much compressed. The epidermis showed focal basal cell vacuolation with associated acute inflammation as early as one hour postexposure. Coagulative necrosis of the epidermis and papillary dermis was complete by 24 hours and followed the appearance of multiple coalescent blisters between six and 12 hours post-exposure. At 48 hours, the lesions were full thickness burns with necrosis extending into the deep subcutaneous connective and adipose tissues. The study of lesions beyond 24 hours revealed early epithelial regeneration at the wound edge. The overall spontaneous healing rate of these biologically severe lesions was significantly faster than comparable sulphur mustard injuries and probably reflected a lack of alkylation of DNA and RNA. PMID- 10365089 TI - The E-cadherin/epidermal growth factor receptor interaction: a hypothesis of reciprocal and reversible control of intercellular adhesion and cell proliferation. AB - The E-cadherin/catenin complex is a calcium-dependent cell-cell adhesion molecule, whose function is critical to the integrity of the adherens junction and which plays a role in the establishment and maintenance of normal epithelial morphology and differentiation. Loss of E-cadherin-mediated adhesion appears to be a fundamental aspect of the neoplastic phenotype which in some cases appears to be mediated by post-translational modifications (i.e. tyrosine phosphorylation) of its interacting proteins, the catenins which link E-cadherin to the actin cytoskeleton. There is increasing experimental evidence to suggest that epidermal growth factor receptor tyrosine phosphorylation may lead to the inactivation of the E-cadherin/catenin complex in cancer cells through its interaction with beta- or gamma-catenin in the cytoskeleton. Modulation of epidermal growth factor receptor activity by pharmacological agents has the potential to regulate a variety of cellular processes including adhesion, differentiation, and proliferation. PMID- 10365090 TI - Somatic hypermutation and B-cell malignancies. AB - During a follicle centre response, the immunoglobulin genes are subjected to a hypermutation mechanism which introduces predominantly single base changes, non randomly, into the immunoglobulin V region (IgV) genes. B cells with mutated IgV genes are then selected according to the affinity of the encoded antibody for antigen retained on the follicular dendritic cells, resulting in an increase in the affinity of the humoral response. The identification of mutated immunoglobulin genes has been applied to the study of normal B cells and B-cell lymphomas to determine either follicle centre cell ancestry, or continued influence of the follicle centre microenvironment. Although analysis of mutations in many lymphomas has confirmed previous hypotheses, there have been some surprises, such as the identification of rearranged and mutated IgV genes in Hodgkin's Reed-Sternberg cells. In this mini-review we will examine the characteristics of the hypermutation mechanism and the way in which mutations in IgV genes have been used to study B-cell malignancies. PMID- 10365091 TI - Cadherin-11 is highly expressed in rhabdomyosarcomas and during differentiation of myoblasts in vitro. AB - Rhabdomyosarcomas bear a morphological and genetic resemblance to developing skeletal muscle. Apart from myogenic marker genes (bHLH factors, myosin, actin), cell adhesion molecules such as N-cadherin and N-CAM have been reported to be expressed both in rhabdomyosarcomas and during myogenesis. The present study demonstrates the expression of another cadherin, cadherin-11, in rhabdomyosarcomas and during differentiation of myoblasts in vitro: cadherin-11, a predominantly mesenchymal cell adhesion molecule, is highly expressed in embryonal rhabdomyosarcomas and alveolar rhabdomyosarcomas, which do not bear the Pax-3-FKHR fusion previously described. Cadherin-11 is down-regulated in normal skeletal muscle and after myotube formation in vitro. The results of this study suggest that cadherin-11 might be involved in myogenesis and that rhabdomyosarcomas may re-express or fail to down-regulate cadherin-11. Since alveolar rhabdomyosarcomas bearing the t(2;13) translocation do not express cadherin-11, it is postulated that Pax-3 and cadherin-11 might be linked and involved in the same myogenic pathway. PMID- 10365092 TI - Down-regulation of CD44 standard and variant isoforms during the development and progression of uterine cervical tumours. AB - To clarify the possible role of CD44 in the development or progression of uterine cervical tumours, an immunohistochemical investigation was carried out on 125 cases of cervical intraepithelial neoplasia (CIN), 78 invasive squamous cell carcinomas (ISCC), 61 cervical adenocarcinomas (AC), nine adenosquamous carcinomas (ASq), and 15 carcinomas with co-existent SCC and AC components, as well as 87 samples of normal cervix. A combination of reverse transcription polymerase chain reaction (RT-PCR) and Southern blot hybridization (SBH) was also applied to 16 cervical carcinomas and 24 normal cervical specimens. Immunoreactivity for CD44s, CD44v3, and CD44v6 did not alter during the progression of CIN, while significantly decreased expression was observed in ISCC, associated with invasive features in some tumours. Reduced levels of CD44 expression in AC were also found, compared with normal cervical glandular epithelia. The average immunoreactivity scores for CD44s, CD44v3, and CD44v6 were significantly higher in ISCC than in AC, in line with the RT-PCR/SBH assay results. However, CD44 scores did not correlate with any clinicopathological factors or with survival in ISCC or AC. The ASq and AC CD44 scores were similar, while staining patterns in mixed tumours were dependent on the morphological phenotype, suggesting a close association between CD44 expression and the cell types. The results suggest that whereas CD44 is down-regulated during cervical tumourigenesis, positivity may not be useful as a consistent prognostic indicator. PMID- 10365093 TI - CD44 expression in sinonasal melanomas: is loss of isoform expression associated with advanced tumour stage? AB - The expression of the adhesion molecule CD44 was examined in 14 primary sinonasal melanomas (SMs), aggressive neoplasms with short survival times, as CD44 overexpression has been linked to poor survival in human cancers. Immunohistochemistry was performed on formalin-fixed, paraffin-embedded sections with CD44 isoform-specific monoclonal antibodies to the CD44 standard (s) and variant isoforms (v) v5 and v6. CD44s, v5, and v6 were strongly expressed in a membranous pattern in SM in situ, early invasive SM, and in uninvolved respiratory/squamous epithelium. In invasive SM, membranous CD44s expression was identified in a large proportion of melanoma cells. Membranous staining of CD44v5 and v6 was lost in invasive SM, independently of the histological subtype. Diffuse cytoplasmic staining was observed focally in invasive SM and loss of cytoplasmic expression of CD44v6 and v5 was associated with advanced tumour stage in the linear-by-linear association test (p = 0.042 and 0.066, respectively). CD44s may not be important for malignant transformation, as it is expressed in both benign and malignant melanocytes. Loss of membranous CD44 isoform expression in widely invasive SM suggests that loss of cellular adhesion facilitates matrix and vascular infiltration and dissemination of sinonasal melanoma cells. PMID- 10365094 TI - Lack of class II transactivator causes severe deficiency of HLA-DR expression in small cell lung cancer. AB - Small cell lung cancer (SCLC) is characteristically not associated with tumour infiltrating lymphocytes. Since SCLC has been reported to show marked reduction of class I HLA, the reduced expression has been considered a means of escaping anti-cancer immunity. However, HLA-DR expressed in cancer cells is now known to contribute to anti-cancer immunity. To clarify the difference in HLA-DR expression between SCLC and non-small cell lung cancer (NSCLC), and the mechanism, the expression and the cis- and trans-acting factors involved were investigated. HLA-DR was not immunohistochemically detected in any SCLC and could not be induced by interferon gamma (IFN-gamma) in any SCLC cell line, whereas HLA DR was expressed to varying degrees and was easily induced in NSCLC. SCLC cell lines lacked class II transactivator (CIITA) even after IFN-gamma induction, whereas NSCLC cell lines expressed CIITA. The other class II HLA-specific transcription factors were expressed and genomic DNA of HLA-DR, including the promoter, was conserved well both in SCLC and in NSCLC cell lines. CIITA transfection improved the expression of HLA-DR in SCLC. In conclusion, the lack of CIITA results in severe deficiency of HLA-DR expression in SCLC. Since CIITA has also been reported to induce class I HLA, CIITA transfection might make it possible to establish effective anti-cancer immunotherapy against SCLC through the up-regulation of class I and class II HLA. PMID- 10365095 TI - Cyclin-dependent kinase inhibitor p27Kip1 expression and interaction with other cell cycle-associated proteins in mammary carcinoma. AB - p27, cyclin D1, and retinoblastoma (Rb) protein have been demonstrated using immunohistochemistry in 189 cases of primary breast carcinoma with long-term follow-up. There was a statistically significant association between the expression of p27 and both cyclin D1 and the retinoblastoma gene product (pRb), corresponding to their close interactions in regulating the G1/S transition in the cell cycle. Low levels of p27 were seen in high-grade, rapidly proliferating, oestrogen receptor-negative tumours. In univariate analysis, low p27 expression was associated with a reduced relapse-free and overall survival. In multivariate analysis, p27 was not an independent predictor of survival when either histological grade or proliferative activity (S-phase fraction) was included in the model. When the combined expression of p27 and cyclin D1 was related to survival, patients with high levels of p27, regardless of their cyclin D1 status, did well, whilst those with low p27 had a poor outcome. The only exception, in the latter group, was patients with tumours expressing high levels of cyclin D1, who did as well as the high p27 group. We have shown that in clinical material p27 expression is associated with proliferative activity and while univariate analysis shows it to be a significant indicator of prognosis, this significance is lost in multivariate analysis when traditional prognostic factors are included in the model. The interest in p27 expression in mammary carcinoma lies in its behaviour when examined in combination with other G1 cell cycle regulators. PMID- 10365096 TI - Correlation between p53, c-erbB-2, and topoisomerase II alpha expression, DNA ploidy, hormonal receptor status and proliferation in 356 node-negative breast carcinomas: prognostic implications. AB - Various new prognostic indicators have been identified for mammary carcinomas, but the issue of their significance remains unsettled. The prognostic impact of p53, c-erbB-2, and topoisomerase II alpha expression was investigated in relation to standard prognostic factors for carcinomas of the breast and to the tumour cell growth fraction. Paraffin-embedded specimens of 356 node-negative infiltrating ductal carcinomas were stained immunohistochemically using a polyclonal antiserum to c-erbB-2, and the monoclonal antibodies DO-1 (p53), Ki-S4 (topoisomerase II alpha), and Ki-S5 (Ki-67). The patients were followed for a median duration of 99 months. Both p53 and c-erbB-2 were significantly associated with high tumour grade, large tumour size, DNA aneuploidy, lack of steroid hormone receptors, young age, and increased topoisomerase II alpha and Ki-67 expression levels. The correlation of p53 and c-erbB-2 was not significant. Topoisomerase II alpha and Ki-67 scores closely paralleled each other, indicating that both reflect the proliferative activity of tumour cells. A univariate analysis of overall (OS), specific (SS), and disease-free survival (DFS) revealed all the above-mentioned parameters to be statistically significant except patient age, which was relevant only to overall survival. Multivariate analysis with inclusion of all covariates selected tumour size and proliferation (topoisomerase II alpha and Ki-67) indices as independent predictors of survival in all three models. No additional information was gained by p53 or c-erbB-2. It is concluded that the proliferative activity, as assessed by topoisomerase II alpha or Ki-67 immunostaining, is the most useful indicator of breast cancer prognosis, except for tumour size. PMID- 10365098 TI - Synthesis of basement membrane by gastrointestinal cancer cell lines. AB - Gastrointestinal adenocarcinoma-derived cell lines were studied in order to determine their pattern of expression of basement membrane components and their ability to form a basement membrane. In contrast to the well-preserved expression of laminin beta 2, beta 3, gamma 1, and gamma 2 chain mRNAs, five of eight gastrointestinal cancer cells lacked alpha 3 mRNA. Immunohistochemical and electron microscopic examination of four cell lines transplanted subcutaneously to SCID mice demonstrated the presence of both alpha 3 and alpha 5 chains and the formation of a basal lamina in two cases. The other two cell lines lacked both alpha 3 and alpha 5 chains and could not form a basal lamina, suggesting that this deficiency may be a factor which affects their ability to form a basement membrane. This abnormality might play some role in stromal invasion by tumour cells in gastrointestinal cancer. PMID- 10365097 TI - Clinicopathological implications of parathyroid hormone-related protein in human colorectal tumours. AB - The purpose of the present study was to clarify the relationship of parathyroid hormone-related protein (PTHrP) to the oncogenesis and progression of colorectal adenocarcinoma. A total of 108 colorectal tumours, including 12 adenomas, six adenocarcinomas in adenomas, and 90 adenocarcinomas, were studied. Immunohistochemistry, in situ hybridization, and reverse transcription-polymerase chain reaction (RT-PCR) techniques were used to evaluate the expression of PTHrP. Positivity of immunostaining for PTHrP was defined as highly positive (++), slightly positive (+), and negative (-). None of the adenomas of background non neoplastic mucosal epithelia showed immunostaining of PTHrP. In contrast, PTHrP was expressed in 85 (94.4 per cent) of 90 colorectal adenocarcinomas. Immunoreactivity of PTHrP was greater in poorly differentiated adenocarcinomas than in well-differentiated ones. Furthermore, advancing margins of primary tumours stained more intensely than other sites. Highly positive immunoreactivity of PTHrP, classified by histological invasiveness, was 22.6 per cent within the muscularis propria and 69.5 per cent beyond the muscularis propria. PTHrP expression was significantly correlated with differentiation, depth of invasion, lymphatic invasion, lymph node metastasis, hepatic metastases, and Dukes' classification. In carcinoma, PTHrP mRNA expression was evident in tumour cells by in situ hybridization. PTHrP transcripts were also detected in two resected human colorectal adenocarcinomas by RT-PCR. These findings suggest that PTHrP is related to carcinogenesis, differentiation, progression, and prognosis of colorectal adenocarcinomas. PMID- 10365099 TI - Expression of growth regulatory genes in a SCID mouse-human model of intestinal epithelial regeneration. AB - Analysis of human intestinal epithelial regeneration has been limited. This study has used a novel SCID mouse-human model to test the hypothesis that distinct stages of human intestinal epithelial regeneration are accompanied by differential expression of growth regulatory genes. Disaggregated epithelium, which included crypt cell aggregates, was isolated from human fetal small intestine and transplanted subcutaneously in SCID mice. This method induced a coordinated regeneration response and enabled temporal separation of cell populations at different stages of histogenesis and cytodifferentiation. Graft epithelium was identified using a specific anti-human monoclonal antibody (MAb 5D3) against cytokeratins 8 and 18. Functional epithelial lineages were identified by appropriate markers. Growth regulatory genes relevant to proliferation and apoptosis, including Bcl-2, p53 and Ki67, were assayed at different stages of regeneration. During early regeneration, Bcl-2, p53, and Ki67 were expressed throughout the epithelial compartment. On completion of regeneration, these genes were expressed only in crypt epithelium and were absent from villi. This study has established a novel SCID mouse-human model of intestinal epithelial regeneration. During early regeneration, increased Bcl-2 and Ki67 expression may indicate suppression of apoptosis and enhanced proliferation respectively, consistent with expansion of the stem cell fraction. The p53 gene may influence pathways of differentiation during regeneration, analogous to its role during development. PMID- 10365100 TI - Amplification of the androgen receptor gene is associated with P53 mutation in hormone-refractory recurrent prostate cancer. AB - p53 protein expression of 30 hormone-refractory locally recurrent prostate cancers was compared with their matched untreated primary tumour specimens. In addition, androgen receptor (AR) gene amplification and p53 protein immunostaining were compared. p53 positivity increased during hormonal therapy from 17 per cent of the untreated primary tumours to 40 per cent of the hormone refractory recurrences (p = 0.078). None of the p53-positive primary tumour specimens lost p53 positivity during hormonal therapy. Hormone-refractory recurrences with AR gene amplification more frequently (p = 0.0342) showed positive p53 immunostaining than tumours without AR gene amplification, 75 and 27 per cent, respectively. In summary, this study has shown that a cell clone with P53 mutation seems to be selected for during endocrine therapy and that positive p53 immunostaining correlates with AR gene amplification. These results suggest that inactivation of P53 may lead to genetic instability in a subset of prostate carcinomas enabling them to achieve properties, such as AR gene amplification, that allow them to grow in low levels of androgens and therefore cause tumour progression. PMID- 10365101 TI - Tenascin is differentially expressed in endometrium and endometriosis. AB - Endometriosis is characterized by the presence of functional endometrial tissue outside the uterine cavity, most commonly on the ovary and peritoneum. The aetiology of endometriosis is not understood, although the adhesion of endometrial cells to the extracellular matrix (ECM) would be expected to play a central role in its pathogenesis. The expression of ECM molecules in endometrium and in endometriosis has been investigated using immunohistochemistry and western blotting techniques. The ECM components collagen IV, laminin, vitronectin, and fibronectin had a similar pattern of expression throughout the menstrual cycle in endometrium and endometriosis. Expression of tenascin was elevated in the stroma of the functionalis region of the endometrium during the proliferative stage of the menstrual cycle and in endometriosis. Tenascin expression in endometriosis was not modulated according to the stage of the menstrual cycle. It is concluded that expression of tenascin is strictly regulated in endometrium and may be important in endometrial regeneration and in the pathogenesis of endometriosis. PMID- 10365102 TI - The histopathology of fibrous dysplasia of bone in patients with activating mutations of the Gs alpha gene: site-specific patterns and recurrent histological hallmarks. AB - Gs alpha mutations and histopathology have been analysed in a series of 13 patients with fibrous dysplasia (FD) of bone, including 12 patients with the McCune-Albright syndrome (MAS) and one patient with monostotic FD. Activating mutations (either R201C or R201H) of the gene encoding the alpha subunit of the stimulatory G protein, Gs, were detected in all cases, including the case of monostotic FD, using a variety of techniques [reverse transcription-polymerase chain reaction (RT-PCR) with allele-specific primers, allele-specific oligonucleotide hybridization, and DNA sequencing]. A spectrum of bone lesions associated with such mutations was identified and it was possible to recognize three primary, but distinct, histological patterns, defined here as Chinese writing type, sclerotic/Pagetoid type, and sclerotic/hypercellular type, which are characteristically associated with the axial/appendicular skeleton, cranial bones, or gnathic bones, respectively. Features of FD histopathology were characterized by confocal fluorescence microscopy, which allowed the definition of osteogenic cell shape changes and 'Sharpey fibre bone' as common denominators of all histological subtypes. Defining characteristics of the different subtypes, two of which diverge from standard descriptions of FD and have never been characterized before, were dependent on the amount and structure of bone tissue within the FD lesion. These data emphasize the non-random (site-specific) variability of FD histopathology in patients carrying activating mutations of the Gs alpha gene and provide additional evidence for the occurrence of Gs alpha mutations in cases of FD other than typical MAS. PMID- 10365103 TI - Lymphocyte targeting of the brain in adoptive transfer cryolesion-EAE. AB - Lymphocyte infiltration and microglial activation in experimental autoimmune encephalomyelitis (EAE) are mainly centred on the spinal cord. However, a cryolesion to one cerebral hemisphere (cryolesion-EAE) induces six-fold enhancement of EAE in the cerebral hemispheres and removal of the cervical lymph nodes reduces such enhancement by 40 per cent. This study tests the hypothesis that lymphocytes from donor rats with cryolesion-EAE will selectively target the brain rather than the spinal cord when transferred to naive recipients. Acute EAE was induced in 15 Lewis rats (donors); ten donors received a cryolesion to the left cerebral hemisphere 8 days post-inoculation of antigen and adjuvant. Five rats with EAE received no cryolesion. Lymphocytes from cryolesion-EAE donors or from EAE-only donors were cultured for 72 h in medium containing myelin basic protein and then injected into a total of 21 naive recipients, which were killed 8 days later. The severity of EAE in brains and spinal cords was assessed in immunocytochemically stained sections by quantifying the number of vessels showing lymphocyte cuffs (W3/13 antibody) and the level of MHC class II antigen expression by microglia (OX6 antibody). When compared with recipients of EAE-only donor lymphocytes, the severity of cerebral EAE was increased 2- to 2.6-fold in the recipients of crylesion-EAE donor lymphocytes (p < 0.01); EAE in the spinal cord was reduced. These results suggest that lymphocytes from cryolesion-EAE donors preferentially target the brain in recipient animals in preference to the spinal cord. By analogy with cryolesion-EAE, focal central nervous system (CNS) damage with drainage of auto-antigens to regional lymph nodes in man may play a role in determining the site and timing of initial and recurrent multiple sclerosis lesions. PMID- 10365104 TI - Functional imaging and detection of local recurrence in soft tissue sarcomas by positron emission tomography. AB - BACKGROUND: Besides anatomically-based radiodiagnostics, functional imaging may be used for characterizing soft tissue sarcomas (STS). This study evaluates positron emission tomography (PET) with three different radiotracers (fluorine-18 deoxyglucose [FDG], carbon-11 aminoisobutyric acid [AIB] and oxygen-15-labeled water [O15-water]) for imaging STS and the detection of local recurrence. PATIENTS AND METHODS: 21 patients presented with 9 primary STS, 5 recurrent STS and 10 lesions suspicious for local recurrence (radiological tomography). Standardized uptake values (SUV) were calculated in tumor and normal tissues (muscle/blood vessels). Surgery with pathological examination or follow-up data were referred to as control criteria. RESULTS: All tracers accumulated in STS with no difference between primary and locally recurrent tumors. The uptake in STS was increased compared to muscle (FDG, P < 0.0001; AIB, P = 0.0039; O15 water, P = 0.002) and to blood vessels for FDG (P < 0.0001) as well as AIB (P = 0.0038). Of 10 patients with suspected recurrence, 6 presented neither PET criteria for recurrence nor local failure in the specimens or during follow-up, while 4 cases with positive PET scans were ultimately diagnosed with local failure. CONCLUSION: Besides FDG, AIB and O15-water were shown to accumulate in primary and recurrent STS of different histologic type. A precise differentiation from normal muscle was found for all tracers. FDG and AIB differentiated viable tumor from blood vessels. PET using FDG, AIB and O15-water is suitable for functional imaging of STS and the detection of sarcoma recurrence. PMID- 10365106 TI - Morphological and genetic abnormalities in prediction of recurrence in radically operated colorectal cancer. AB - BACKGROUND: We analyzed clinicopathological variables, cell proliferation activity and genetic aberrations related to colorectal cancer in order to recognize clinically usable predictive markers of cancer recurrence. MATERIALS AND METHODS: A total of 111 patients radically operated upon because of primary colorectal cancer in 1986-1991 were studied. Loss of heterozygosity (LOH) at 18q21 and replication errors were studied by polymerase chain reaction and fragment analysis. Expression of p53 protein and that of Ki-67 were studied using immunohistochemical methods. RESULTS: LOH at 18q21 was the only factor associated with recurrence (P = 0.03), and indicated a worse five-year cumulative survival rate (42%) than did LOH-negativity (72%) in cases of Dukes classes B and C. Expression of p53 protein indicated recurrence (P = 0.07), short disease-free time and poor survival (P = 0.03) in Dukes class A cases. CONCLUSIONS: LOH at 18q21 appears useful in predicting recurrence and poor survival in cases of Dukes classes B and C, as does p53 expression in class A cases. PMID- 10365105 TI - Expression of Bcl-2, Bax, and p53 proteins in carcinogenesis of squamous cell lung cancer. AB - BACKGROUND: Apoptosis is regulated by many genes, including bcl-2, p53, and bcl-2 family genes. The expression of Bcl-2, p53, and Bax in very early lung cancers or precancer lesions is not been fully understood. MATERIALS AND METHODS: The expression of these proteins was examined immunohistochemically in 11 normal bronchial epithelia, 23 dysplasias, and 40 roentgenographically occult squamous cell lung cancers (ROCs). RESULTS: The expression of the Bcl-2 and p53 protein increased along with the advance of the morphological atypia in bronchial epithelial cells, whereas no difference in the Bax expression. The patients with Bcl-2 positive ROCs had a better prognosis than those with Bcl-2 negative ROCs. CONCLUSIONS: Bcl-2 and p53 proteins play important roles in early steps of carcinogenesis in squamous cell lung cancer of central type. The Bcl-2 protein could be a prognostic marker in ROCs. PMID- 10365107 TI - Evaluation of the CA 242 tumor antigen as a potential serum marker for colorectal cancer. AB - OBJECTIVE: The present study was designed to define the performance of serum CA 242 as a marker in colorectal cancer patients. PATIENTS AND METHODS: Serum samples from 1,013 subjects (440 healthy volunteers, 384 patients with primary or recurrent colorectal carcinoma and 189 with benign colorectal diseases) were evaluated. RESULTS: The measurement of serum CA 242 levels in the population of healthy subjects demonstrated the presence of positive levels in approximately 5% of the cases. Interestingly, similar results (5.8%) were obtained in patients with benign colorectal disease, demonstrating the high specificity of CA 242. When serum samples from colorectal cancer patients were analyzed, a sensitivity of 34.9% was observed. Moreover, 18.6% Stage A and B patients had positive CA 242 levels, compared to 33.3% and 58.8% of Stage C and D patients, respectively, indicating a correlation with the stage of disease. A comparison between preoperative and immediate postoperative CA 242 levels showed a consistent relationship between the efficacy of surgery and the reduction in serum CA 242 levels; further, elevated CA 242 levels were present in the immediate postsurgical follow-up of patients undergoing palliative surgery. A longitudinal evaluation of serum CA 242 levels demonstrated that this marker was indicative of the status of disease. CONCLUSIONS: The results obtained suggest the possible utility of CA 242 in monitoring the disease status, providing a rationale for future studies focusing on the longitudinal monitoring of colorectal cancer patients. PMID- 10365108 TI - Clinical investigation of bronchioloalveolar carcinoma: a retrospective analysis of 53 patients in a single institution. AB - BACKGROUND: Bronchioloalveolar carcinoma (BAC) has been reported to have unique clinicopathological features. PATIENTS AND METHODS: This retrospective study was performed using data base including 871 patients treated for primary lung cancer between 1981 and 1995. RESULTS: The patients with BAC included a larger proportion of female (P = 0.029) and smoked less (P = 0.002) than those with non BAC. There was no difference in survival between surgically resected patients with BAC and those with non-BAC. Clinical Stage IV patients with BAC had a better response to chemotherapy than did those with non-BAC. Survival in the former group was better than that in the latter on univariate analysis, but the significance of this difference was not confirmed multivariate analysis. CONCLUSION: The patients with BAC included a larger proportion of females and smoked less than those with non-BAC. Treatment results for BAC was comparable to those for non-BAC. PMID- 10365109 TI - Intraperitoneal chemo-hyperthermia with mitomycin C for gastric cancer patients with peritoneal carcinomatosis. AB - OBJECTIVE: This study evaluates the tolerance and efficacy of Intraperitoneal Chemo-hyperthermia (IPCH) with Mitomycin C (MMC) associated with surgery, in peritoneal carcinomatosis of gastric origin. BACKGROUND: Most patients with peritoneal carcinomatosis of gastric origin die within 6 months, and IPCH associated with surgery has been reported as a possible new therapeutic approach. METHODS: A prospective non randomized trial was carried out on 42 patients with gastric cancers and peritoneal carcinomatosis. Fourty-three IPCH with MMC were used as complementary treatment after surgery (peritoneal perfusate with a 10 mg/l dose of MMC, inflow temperature 46 to 49 degrees C, use of a closed circuit, duration 90 minutes). Fourteen primary tumors were unresectable ones and 12 patients had large malignant preoperative ascites. RESULTS: Mortality and morbidity rates were 2/42 and 4/42 respectively. For resectable gastric cancers with stage 1 and 2 carcinomatosis (malignant granulations less than 5 mm in diameter), one, two and three year survival rates were 80, 61 and 41% respectively. For unresectable primary tumors and for stage 3 and 4 carcinomatosis (granulations larger than 5 mm in diameter), six and twelve month survival rates were 50% and 10% respectively. CONCLUSIONS: IPCH appears as a safe new therapeutic approach in gastric cancers with peritoneal carcinomatosis with small malignant granulations (stage 1 and 2) and randomized trials are now needed to clearly evaluate its efficacy. PMID- 10365110 TI - Prognostic factors in ambulatory patients with inoperable locoregionally recurrent rectal cancer following curative surgery. AB - BACKGROUND: The optimal treatment for locoregionally recurrent rectal cancer after curative surgery has not yet been defined. The definition of prognostic factors could lead to the selection of an aggressive therapeutic approach in patients with favourable prognosis alone. PATIENTS AND METHODS: The records of thirty-nine ambulatory pts, 15 female and 24 male, with diagnosis of locoregionally recurrent rectal cancer (LRRC) after curative surgery and treated with radiotherapy were retrospectively analyzed. The following factors were analyzed for their ability to predict the clinical response and outcome for LRRC: age, sex, initial tumor grading, primary surgical approach, initial primary tumor stage according to Dukes' classification, disease free survival (time to primary surgery and detection of a LRRC), pelvic-perineal structure affected by recurrence, total radiation dose, chemotherapy with fluorouracil, symptomatic response to the therapy, locoregional symptomatic re-recurrence, systemic progression disease. RESULTS: In the univariate analysis, predictive factors for survival, were graded (G1-2 vs G3 p = 0.04), Dukes' stage at first diagnosis (A-B vs C p = 0.01), and site of pelvic-perineal recurrence (Pelvic mass alone yes vs no p = 0.01; Nerve and/or Osseous involvement yes vs no p < 0.001). Following therapy for LRRC, a better survival was observed in pts with a complete symptomatic response (complete remission vs partial remission vs no change p < 0.001), without a further locoregional symptomatic re-recurrence (re-recurrence, yes vs no p = 0.001) and/or appearance of metastatic disease (yes vs no p < 0.001). PMID- 10365111 TI - Human papilloma virus DNA detection in oral lesions in the Greek population. AB - HPVs (Human Papilloma Viruses) are small DNA "epitheliotropic" viruses, implicated in cervical carcinogenesis, particularly the "high-oncogenic-risk" types HPV-16 and HPV-18. Data concerning oral carcinogenesis are however, contradictory. We examined the presence of HPV and subsequently HPV-16 and HPV-18 in 102 specimens of paraffin-embedded oral tissue blocks--81 oral squamous cell carcinomas and 21 oral hyperplasias--using PCR technique followed by agarose gel electrophoresis. Our results demonstrated that 49% (50/102) of the samples were HPV positive. Subsequent analysis of HPV positive lesions revealed 22% positivity for HPV-16 and 44% for HPV-18. HPV-18 was detected only in carcinomas, while HPV 16 was more abundant in papillomatous hyperplasias and in a small percentage of carcinomas. These findings may probably indicate a contributing role for HPV-18 as a potent co-carcinogen in oral epithelial carcinogenesis in the Greek population. PMID- 10365112 TI - Decreased serum levels of insulin-like growth factor (IGF)-I in patients with lung cancer: temporal relationship with growth hormone (GH) levels. AB - AIMS AND BACKGROUND: Several studies have evidenced that IGF-1 may play a role in the growth regulation of many cancer cell lines, and recently GH and IGF-1 have been recognized as stimulators of lymphopoiesis and immune function. We investigated whether there are differences among health- old people and old people suffering from lung cancer at different stages of disease in the 24-hour secretory profiles of GH und IGF-1. METHODS: The study was carried out on seven healthy volunteers (mean age +/- s.e. 68.8 + 1.92), seven patients with I and II stage lung cancer (mean age +/- s.e. 67.2 +/- 0.80) and seven patients with III and IV stage lung cancer (mean age +/- s.e. 69.5 +/- 2.26). GH and IGF-1 serum levels were measured on blood samples collected every four hours for 24 hours; the area under the curve (AUC) and the presence of circadian rhythmicity were evaluated. RESULTS: A normal circadian rhythmicity was recognizable only for GH secretion in healthy subjects. A progressive increase of GH serum levels and a steady decrease of IGF-1 serum levels were observed in cancer patients in relation to advancing stage of neoplastic disease. CONCLUSIONS: Lung cancer is associated with an altered regulation of GH-IGF-1 system, that might play a role in the clinical course of neoplastic disease. PMID- 10365113 TI - Serum preoperative vascular endothelial growth factor (VEGF) in epithelial ovarian cancer: relationship with prognostic variables and clinical outcome. AB - Substantial experimental and clinical evidence links tumor growth, progression and metastatic potential with neoangiogenesis. This process is modulated by several angiogenic growth factors, such as vascular endothelial growth factor (VEGF). Little data are currently available on serum VEGF levels in cancer patients. In the present retrospective investigation preoperative serum VEGF was higher in 53 patients with epithelial ovarian cancer than in 25 patients with benign ovarian disease as controls (median, range: 229.7, 23.5-1807.5 pg/ml versus 140.3, 14.7-1038.7 pg/ml, p = 0.034). With regard to FIGO stage, antigen values were significantly elevated in stage III-IV (p = 0.027) but not in stage I II ovarian cancer patients when compared to controls. In patients with advanced disease preoperative serum VEGE was significantly related to the presence of ascites (p = 0.013), but not to common prognostic variables, response to chemotherapy and survival. In conclusion, preoperative serum VEGF assay reflects tumor progression and ascites generation in epithelial ovarian cancer, but it seems to have a limited predictive and prognostic value in patients with advanced disease. PMID- 10365114 TI - 5-Fluorouracil plus high dose levofolinic acid and oral hydroxyurea for the treatment of primary hepatocellular carcinomas: results of a phase II multicenter study of the Southern Italy Oncology Group (G.O.I.M.). AB - A phase II trial of 5FU in modulation with intravenous high-dose levofolinic acid and oral hydroxyurea (HU) in advanced unresectable hepatocellular carcinoma (HCC). A total of 50 consecutive patients, 38 males (76%) and 12 females (24%), with a mean age of 62 years (range 30-74) and a mean performance status of 80 (KI, range 60-90) were enrolled. The vast majority of patients were therapy naive, although two patients (4%) had previous surgery and showed progressive disease at entry. No patient had been previously treated with chemotherapy. Five patients had previous hormonotherapy with tamoxifen. Most patients had disease limited to the liver while 12 patients (24%) had also metastatic deposits outside the liver. The treatment plan included: levofolinic acid 100 mg/m2 diluted in 500 cc of normal saline over 2 hour infusion followed by 5FU 600 mg/m2 i.v. bolus. HU 1,000 mg/m2 was given by mouth in three refracted doses starting 6 hours after 5FU. A PR was recorded in only 5 patients (10%; 95% CL 1%-34%) with a median duration of 5.7+ months (range 4.0/6.2 months), a stabilization in 15 (30%) with a median duration of 3.8 months, while 30 patients progressed (60%). PR were seen at liver primary tumor in 4 cases and at soft tissue in 1 case. The median survival was 5.8 months (range 2.0/12.0+). The most frequent toxicities were leukopenia (32%), which however was mild (grade 1-2) in all cases, and grade 1-2 thrombocytopenia observed in 15% of cases. Mild grade 1-2 vomiting was recorded in one third of patients, and grade 1-2 stomatitis in 15%. The combination of 5FU with levofolinic acid and oral HU on a weekly schedule is largely inactive against unresectable or metastatic HCC and results are no better than historical data reported for 5FU alone. PMID- 10365115 TI - Unwinding Ariadne's thread: some methodological and interpretative considerations on international hospital admission comparisons, with special reference to cancer. AB - Hospital utilisation rates comprise the main available source of information on morbidity patterns and the need for health care in a number of developing countries. On the basis of the findings of a Greek-Swedish comparison of somatic hospital admission (presented per the 18 principal head groups of the International Classification of Diseases) between two regional University hospitals, a number of important findings and methodological questions are considered. Factors that can explain the large variations of hospital admission found are examined; in particular those related to disease definition, classification and coding, hospital services supply, professional behavior and practices, illness behavior of patients, and demographic and socio-economic characteristics but especially a true difference in morbidity. An additional statistical elaboration of the hospitalization data in the form of a logistic regression analysis is suggested, in particular at the district or regional level. However, after rigorous scrutiny, our data point to persistent remarkable differences at the respective sites in the major "welfare-related" population diseases, notably, cancer and cardiovascular diseases. PMID- 10365116 TI - DNA aneuploidy by flow cytometry is an independent prognostic factor in squamous cell carcinoma of the oral cavity. AB - BACKGROUND: The clinical significance of flow cytometric DNA content in head and neck squamous cell carcinoma is still controversial. PATIENTS AND METHODS: We prospectively investigated the prognostic importance of tumor size (T), lymph node involvement at presentation (N), degree of histologic differentiation (G) and DNA ploidy (DNA) in 429 surgically treated patients with primary oral squamous cell carcinoma using multivariate statistics. RESULTS: T, G and DNA were independent predictors of metastasis to the neck. N and DNA were independent prognostic factors of loco-regional recurrence development. N and T were prognostic of overall survival. N and DNA were predictive of recurrence-free survival. For the neck negative group, DNA remained as the only predictor of overall and relapse-free survival. CONCLUSION: DNA flow cytometry adds significant prognostic information beyond that provided by established prognostic factors. PMID- 10365117 TI - Nucleolar protein p120 expression in oral carcinoma. AB - Nucleolar protein p120 is a proliferation-associated antigen expressed by cells in early G1 phase, identified by the monoclonal antibody FB-2. Its expression has been evaluated in breast and prostate cancer, and proved to be significantly correlated with other prognostic parameters. In oral pathology, p120 protein content is able to distinguish between non-neoplastic and malignant lesions. In the present study, the immunohistochemical expression of p120 protein was evaluated in fifty cases of oral squamous carcinoma and compared with histological grading, pTNM staging, DNA ploidy status and follow-up of the patients, in order to establish its prognostic value, p120 mean area was significantly correlated to all these parameters, apart from lymph node involvement, indicating the strong predictive potential of this marker. In conclusion, quantitative immunohistochemical analysis of p120 protein represents an easy and reliable method for the assessment of clinical outcome and the definition of risk groups in oral carcinoma. PMID- 10365118 TI - Serum interleukin-6 levels correlate to tumor progression and prognosis in metastatic breast carcinoma. AB - Serum concentrations of tumor necrosis factor-alpha (TNF-alpha), interleukin (IL) 1 beta, IL-2 and IL-6 were determined by enzyme-linked immunosorbent assay or chemiluminescent enzyme immunoassay in 46 Japanese patients with metastatic breast cancer. No IL-2 activity was detectable, only two patients had a detectable TNF-alpha level, and 4 had detectable IL-1 beta concentrations. No correlations were found between TNF-alpha and IL-1 beta levels and clinicopathological parameters. Twenty-two patients (48%) showed higher serum IL 6 levels than the cut-off value of 4 pg/ml. Significantly higher IL-6 levels were detected in patients with more than one metastatic site and with dominant metastatic visceral disease than in those with one metastatic site (median, 6.9 vs 1.1 pg/ml, P < 0.0001) or with dominant metastatic bone or soft tissue disease (median, 7.1 vs 2.4, P < 0.05). The patients with liver metastasis and pleural effusion showed significantly higher serum IL-6 levels than those without liver metastases (median, 17.2 vs 2 pg/ml, P = 0.0006) or pleural effusion (median, 12.1 vs 2.1 pg/ml, P = 0.02). A strong correlation was observed between IL-6 levels and C-reactive protein levels (r = 0.47, P = 0.0006). Patients unresponsive to chemo-endocrine therapy showed significantly higher serum IL-6 levels than those who responded to chemo-endocrine therapy (P = 0.0007). Moreover, the patients with high IL-6 levels showed significantly poorer survival than patients with low IL-6 levels. Multivariate analysis revealed that IL-6, as well as disease-free interval, is an independent prognostic factor of metastatic breast cancer. These results suggest that IL-6 levels are elevated and may be an aggressive parameter in patients with metastatic breast cancer. Higher IL-6 serum levels are found to predict a poorer response to chemo-endocrine therapy, and to represent a poorer prognostic predictor in metastatic breast cancer. PMID- 10365119 TI - Long-term results of surgical treatment for invasive thymoma. AB - BACKGROUND: The aim of this study was to analyse the operative outcome of extensive surgery for invasive thymoma and to assess the prognostic factors for long-term survival. MATERIALS AND METHODS: Forty patients with invasive thymoma have been operated on at our institution during past the 33 years. We performed total removal of the tumour, including invaded neighboring organs. Complete resection was performed in 27 patients, incomplete resection in 4. Nine patients had unresectable thymoma. Postoperative radiotherapy was performed in 30 patients with a median dose of 48 Gy. RESULTS: The 10-year survival rate was 72% for Masaoka stage II, 47% for stage III, and 0% for stage IV. There was no postoperative mortality. Concerning the prognostic factors for long-term survival, there were no significant differences in the analysis of Masaoka staging, histological classification, association of autoimmune disease, and postradiotherapy. However, the survival rate was significantly higher for patients with complete resection than for patients with incomplete resection or biopsy only (p = 0.019). CONCLUSIONS: Whether the tumour is resected completely or not is the most important factor for long-term survival; therefore it is preferable to perform extensive surgery for invasive thymoma. PMID- 10365120 TI - Antiblastic chemotherapy in the presence of abdominal aorta aneurysm (AAA): guidelines. AB - Concomitant neoplasm and abdominal aortic aneurysm (AAA) is not a rare clinical association and can pose problems if antiblastic treatment is required. The literature shows lack of consensus owing to the fact that AAA can modify the liquid load and the hemodynamic setting so that chemotherapy toxicity profile becomes important and hydration overload increases the AAA-breakage risk. We have analyzed the possibility of treatment with a non-critical water load related chemotherapy in these patients, and if it can offer benefit it terms of overall survival (OS) and quality of life (QL). We concluded that in chemotherapy the presence of AAA does not have to be excluded first, if such critical parameters such as response to chemotherapy AAA-breakage risk vascular and extravascular toxicity do not compromise the OS and QL of the patients. PMID- 10365121 TI - Serum cholecystokinin and neurotensin during follow-up of pancreas, prostate and medullary thyroid tumors. AB - BACKGROUND: Growth of pancreatic carcinoma cells is stimulated by cholecystokinin (CCK) and neurotensin (NT). Prostatic carcinoma cells can secrete neurotensin. The CCK gene has been described in thyroid medullary carcinomas (MCT). METHODS: Serum CCK and NT were measured by RIAs during monitoring of 19 pancreas tumours, 10 prostate adenocarcinomas and 10 thyroid medullary cancers (MCT). RESULTS: No correlations were found between CCK and NT in the three tumour types, nor with CA 19.9, PSA, CEA or calcitonin. In pancreas adenocarcinomas (n = 12), initial median CCK was > 8pg/ml (non significant differences between stages T, N or M). Median NT was > 80 pg/ml in all but M0 and stage I-II cases, and significantly higher in M1 and stages IV (P = 0.002). Non significant differences were found for CCK and NT according to clinical stages. In prostate cancers, median CCK was significantly more elevated after relapse (P = 0.040). Median NT was significantly more elevated in disease-free patients (P = 0.04). In MCT, CCK and NT were not related to clinical stages. CONCLUSION: In pancreas and prostate cancers serum CCK may follow tumour load and disease progression. NT was lower in progressive disease. The contribution of these peptides in human tumour growth, since they may have therapeutic implication, warrants further investigation. PMID- 10365122 TI - Loss of heterozygosity of WT1 gene in the prognosis of sporadic Wilms' tumour in children. AB - The aim of this report was to evaluate the prognostic value of allele loss of the WT1 gene in children with sporadic Wilms' tumour. Allele loss of the WT1 gene was evaluated using microsatellite polymorphisms in the 3' untranslated region of WT1 in a radioactive PCR assay. The study comprised 66 children (30 girls and 36 boys), aged from 2 days to 13 years, treated for Wilms' tumour according to the SIOP-09 and PGGL scheme. We have used DNA isolated from the neoplastic versus normal kidney tissue from the paraffin embedded sections using microdissection procedure. Loss of heterozygosity (LOH) of the WT1 gene was found in 12 children (19.6%), 5 cases were non-informative. No significant correlation could be found between the LOH of WT1 gene and sex and age. Significantly more frequent occurrence of LOH in tumor in low stage of advancement and low degree of malignancy was found. However, no significant effect of LOH of WT1 gene was observed on frequency of recurrences, metastasis and deaths. Study of allele loss of the WT1 gene may be recommended in difficult cases as an additional factor useful for the diagnosis and in the assignment of the tumour to the appropriate risk group. PMID- 10365123 TI - High-dose chemotherapy with hematopoietic stem cell support in patients with advanced epithelial ovarian cancer: analysis of 67 patients treated in a single institution. AB - BACKGROUND: Advanced ovarian carcinoma is a chemosensitive tumor, but its prognosis is poor with 20 to 30% 5-year survival using conventional therapy. Increasing doses of chemotherapy might improve the prognosis because of the dose effect. MATERIALS AND METHODS: Between 1980 and 1994 at Institut Paoli-Calmettes, 67 patients with advanced ovarian cancer were treated with different alkylating agents-based regimens of high dose chemotherapy (HDC) and hematopoietic stem cell support (HSCS). The population was divided in 2 groups: a salvage group (n = 30) including initial conventional chemotherapy-refractory patients, and a consolidation group (n = 37) including patients whose disease was sensitive to classical first-line chemotherapy after debulking surgery. RESULTS: Toxicity was essentially hematological (severe aplasia) and digestive (mucositis). Four toxic deaths occurred related to infection during immunosuppression. In the salvage group, 9 out of 22 evaluable patients responded (41%), but the duration of responses was short (median range of 6 months) and the 2-year overall survival rate was 13%. In the consolidation group, 17 patients are still alive, 12 with progression with a median follow-up of 63 months. The 5-year disease-free survival rate was 32% while the 5-year overall survival rate was 46%. CONCLUSIONS: Toxicity of HDC with HSCS is acceptable for poor-prognosis patients. In the long term, this therapy is not beneficial for chemotherapy refractory patients, despite objective response rates better than the conventional treatment, confirming the dose-effect for alkylating agents in ovarian cancer. On the other hand, the results seem better than classical therapy in case of chemosensitive disease and should be confirmed prospectively in a larger cohort of patients. PMID- 10365124 TI - The problematic nature of metastasized renal cell carcinoma. AB - The treatment of metastatic renal cell carcinoma presents major unsolved problems. All of the therapeutic options have shown only minimal success rates. In addition to partial clarification of tumour genesis, basic findings regarding the heterogeneity of tumors were made. Options for further therapeutic developments will result from increased knowledge of the pathogenesis of metastatic spread. After the growth of new tumor vessels (angiogenesis) the metastasizing cell must break away from the cell formation (loss of cadherines). By migration it reaches the vessel wall which is made permeable by proteolysis (matrix metaloproteinase). After reaching the target organ, adhesion results (adhesion molecules and integrins) within the vessel system and ex-travasation follows. With stimulation the metastatic cell will grow in the target organ. Growth is subject to the cytokine control mechanism (interleukines). Based on these individual steps, future therapeutic options can be developed. However, the treatment modalities at our disposal today must not be neglected: for example, immuno(chemo)therapy and various radiation therapies as well as metastases surgery. PMID- 10365125 TI - Assessing genetic heterogeneity of renal cell tumors. AB - Renal cell tumors display a highly variable morphology which is also reflected at the genomic level. Such heterogeneity was at first monitored by cytogenetic means (numerical and structural chromosomal aberrations); in the meantime, more refined molecular techniques allow the assessment of DNA losses or gains in metaphase chromosomes or tissue sections. Moreover, genomic instability can be monitored using microsatellite probes. All these methods document specific characteristics of certain renal cell tumor types, e.g. telomeric associations in chromophobe carcinomas or oncocytomas, typical losses in 3p in clear cell carcinomas or trisomy 7 in renal cell adenomas and carcinomas. Next to examples demonstrating these alterations the Heidelberg classification of renal cell tumors that is based on genomic observations is discussed. PMID- 10365126 TI - Flow cytometric analysis of DNA-aneuploidy subgroups and proliferation in renal cell carcinoma. AB - BACKGROUND: The course of patients suffering from renal cell carcinoma varies considerably and cannot be predicted by tumor stage and grade alone. However, it is crucial to select patients with high risk of progression and to commence adjuvant immuno-chemotherapy in good time. MATERIALS AND METHODS: Multiple samples of 71 kidney tumors were studied by DNA flow cytometry. Aneuploidy was classified into subgroups employing the DNA-index. In tumors of euploid pattern and corresponding normal tissue cell cycle analysis was performed. RESULTS: 39% of tumors were found to be aneuploid. Mean proliferation fraction was distinctly higher in euploid tumors (15.6%) than in normal tissue (6.1%). DNA ploidy pattern correlated significantly (p < 0.05) with histological grading. With increasing tumor size the clonal spectrum changed as well: Tetraploid cell lines fell from 40% to 28%. The number of triploid clones rose from 33% to 56%. CONCLUSION: Based on selection of tri- and hypertetraploid carcinomas, a high-risk-group for tumor recurrence can be associated within the predominating T2/3 G2 kidney tumors. The aim is to treat these patients following curative surgery at the stage of probable micro-metastases while keeping risk of overtreatment as low as possible. PMID- 10365127 TI - Loss of chromosomes in clear cell renal cell carcinoma and in corresponding renal parenchyma. AB - Chromosome studies were done on six renal cell carcinomas (RCC) and on the corresponding renal parenchymas of the tumor bearing kidneys. Histopathologically, all tumors belonged to the clear cell subtype. All examined parenchymas were pathologically benign. None of the tumor cells showed the typical chromosomal aberrations described for (nonpapillary) RCC, i.e. deletions in the short arm of chromosome #3, or gains in the long arm of chromosome #5. In our series both the tumor and the benign kidney tissues were characterized by loss of chromosomes, especially of the chromosomes #6, #9, #16, #20, and of the Y chromosome. Trisomy of chromosome #7 was found frequently in benign parenchyma cells. The identical chromosomal changes in the tumor and in the parenchyma tissues might reflect rather in vivo mosaics rather than primary chromosomal aberrations in the oncogenetic process of clear cell RCC. PMID- 10365129 TI - Differentiation of multifocal renal cell carcinoma by comparative genomic hybridization. AB - Multifocality occurs in 12-22% of all renal cell carcinoma (RCC) cases. In order to differentiate multifocal RCC, we performed comparative genomic hybridization (CGH) and correlated the results with histopathological and clinical data. After histopathological examination of multifocal tumors, tumor areas were dissected from histopathological sections. Isolated DNA was amplified by DOP-PCR. CGH was performed according to standard protocols. Twenty cases were analyzed. In eight cases, at least one identical aberration was detected in the related tumors. Another eight cases showed different chromosomal changes in tumors of the same kidney. In the small additional clear cell tumors typical genetic alterations of clear cell carcinomas were detected whereas in the chromophilic tumors aberrations characterizing both adenomas and carcinomas were found. From these results we conclude that multifocal RCC represents a heterogeneous group of tumors. The malignant potential of small additional tumors in multifocal RCC cannot be excluded. PMID- 10365128 TI - DNA cytophotometry in renal cell carcinoma: a significant prognostic factor? AB - We report on a prospective DNA cytophotometric study of 66 patients with renal tumors, 61 of whom had renal cell carcinoma (RCC) (pT1-pT4, G1-G3). 16 of the patients had a metastasis at the time of diagnosis. Cell material from 1-5 specimens of each tumor was analyzed for intratumoral heterogeneity. The aim of the study was to evaluate the prognostic value of the following DNA parameters: DNA ploidy, DNA grade of malignancy (DNA MG), mean DNA, DNA index, 2c deviation index (2cDI), and 5c exceeding rate (5cER). In this study 21% of the tumors were non-aneuploid, 79% were aneuploid; however, it proved possible to diagnose 38% of the total collective as aneuploid only by analyzing several tumor areas. In five of 61 RCC patients who died during an observation period of 42 months, at least one area of the primary tumor was aneuploid. Aneuploid primary tumors also accompanied the development of metastases and recurrent tumors in four of the 61 RCC patients. Only DNA 2cDI was found to have a significantly positive clinical correlation with metastasis (r = 0.261) during the clinical course. This was not true, however, for the histopathologic parameters. Significantly positive correlations were found between the tumor stages and the following DNA parameters: mean DNA, DNA index, and 5cER. Histopathologic tumor grading showed a significantly positive correlation with DNA MG, mean DNA, and 5cER. Statistically, the mean values of all evaluated parameters were significantly higher in metastasizing and recurrent RCCs than in non-metastasizing carcinomas (p < 0.05; t-test). DNA cytophotometry cannot substitute histopathologic prognosis. However, the analysis of various DNA parameters helps considerably in evaluating both the malignant potential of kidney tumors and the benign parenchyma of tumor-bearing kidneys. PMID- 10365130 TI - Adjuvant interferon alpha therapy in renal cell carcinoma (RCC): prognostic value of DNA cytophotometry. AB - BACKGROUND: Adjuvant immunotherapy with interferon alpha in non-metastatic RCC is still controversial. It was the aim of the present study to investigate whether tumor ploidy can help to isolate a group of patients profitting from this treatment. MATERIALS AND METHODS: Survival data of 119 patients undergoing tumor nephrectomy because of a non-metastatic RCC were analyzed retrospectively. Ploidy was measured in every tumor by means of DNA-cytophotometry. 24 patients received an adjuvant therapy with interferon alpha for one year. Statistical evaluation was performed by the Kaplan-Meier-Method with the logrank-test. RESULTS: Ploidy could be measured in all tumors. 56 (47%) tumors showed a diploid, 63 (53%) an aneuploid DNA-distribution. Ploidy was seen to be a good prognostic factor in RCC with significantly better survival in patients with diploid than those with aneuploid tumors. The adjuvant interferon therapy provided a marginal however not significant better survival in diploid tumors. Patients with aneuploid RCC had no benefit from this treatment. CONCLUSIONS: The data show that an interferon treatment in non-metastatic RCC might improve survival in patients with the prognostic better diploid tumors. However further investigations are necessary to give a general recommendation for this therapy in this group of patients. Additional molecular tumor factors like S-phase analysis and others might also improve patient selection. PMID- 10365131 TI - The histopathological heterogeneity of small renal cell carcinoma. AB - BACKGROUND: Renal tumors resembling renal cell carcinoma but less than 3 cm in diameter historically have been regarded as adenomas because of their low frequency of metastases. However, this concept has been challenged, and it seems that all of these lesions should be considered carcinomas. Thus, the extent of radical surgery of these findings have been reconsidered, in view of the uncertainty regarding their malignant or benign nature. MATERIALS AND METHODS: 107 tumors 40 mm or less in diameter were accordingly divided into three groups and clinico- and histopathological criteria were correlated: group 1: 20 mm or less (n = 33), group 2: 21-30 mm (n = 28) and group 3: 31-40 mm (n = 43). RESULTS: Both lymph node metastases and distant metastases were well correlated with tumor size. Grade 1 renal cell carcinomas decreased in their frequency with an increasing tumor diameter. In grade 3 carcinomas an opposite result was found. With an increase of tumor size the frequency of venous involvement increases as well. Significant more multifocal malignant renal cell carcinoma were seen in renal cell carcinoma between 21-40 mm compared to tumors 20 mm or less in diameter. CONCLUSION: Although the metastatic potential and the biology of small renal tumors are not yet known, it seems that nephron-sparing surgery in patients with renal cell carcinoma more than 20 mm in diameter should only be performed when there is an absolute indication, such as bilateral carcinomas, single kidney or renal failure. The problem is that a long-term follow-up study is mandatory to justify partial nephrectomy as a nephron-sparing operation for renal cell carcinoma more than 20 mm in patients with normal function of the contralateral kidney. PMID- 10365132 TI - Cadherins in renal cell carcinoma. AB - Distant metastasis is the most predominant clinical problem in renal cell carcinoma. The first step of metastasis is detachment of tumor cells from the primary tumor and subsequent access to the circulation e.g. to lymph or blood vessels. Conceptually, detachment of tumor cells may be facilitated by loss of molecules that provide adhesion of normal cells, such as the cadherins. It has in fact been demonstrated that loss of E-cadherin is associated with invasion and metastasis in animal models and with an unfavorable prognosis in cancer patients. Four major cadherins have been demonstrated in normal kidney and renal cell carcinoma: N-cadherin, E-cadherin, ksp-cadherin, and cadherin 6. The particular role of each of these cadherins in pathogenesis of renal cell carcinoma is not completely understood at present. PMID- 10365133 TI - Formulation of interleukin-2 and interferon-alpha containing ultradeformable carriers for potential transdermal application. AB - INTRODUCTION: Transfersomes (TF) are new highly deformable hydrophilic lipid vesicles, which are able to spontaneously penetrate the skin barrier because of their characteristics. Transfersomes are able to transport non-invasively low as well as high molecular weight molecules into the body. We describe the formulation and several biological characteristics of Interleukin-2 and Interferon-a containing TF. MATERIAL AND METHODS: TF contain natural phosphatidylcholine and sodium cholate. Recombinant human IL-2 and human hybrid interferon-alpha A/D were added to TF and incubated for 24 hours at 4 degrees C. Immunotransfersomes were isolated from free IL-2 and IFN by filtration (Centrisart, Sartorius). Biological activity of immunotransfersomes was measured by CTLL-cell-assay for IL-2 and by A549--EMCV-assay for IFN, concentrations of proteins by ELISA. RESULTS: It has been possible to incorporate a high amount of IL-2 and IFN in TF (75-80%). Incorporated IL-2 and IFN were biological active. The increase of the proportion of lipid to protein to 90.9/1 led to growing probability of association. CONCLUSION: We were able to show, that IL-2 as well as IFN is trapped by transfersomes in biological active form and in sufficient concentrations for immunotherapy. In upcoming experiments these IL-2 and IFN containing TF are used for a transdermal approach in the murine RENCA cell line model. PMID- 10365134 TI - Altered expression of beta 1 integrins in renal carcinoma cell lines exposed to vinblastine. AB - BACKGROUND: Cellular expression of P-glycoprotein (P-gp) which mediates a well characterized mechanism of multidrug resistance (MDR) has been reported previously to be associated with an enhanced tumor dissemination. Since adhesion receptors of the beta 1 integrin family play a substantial role in tumor spread, we studied expression of VLA-1 to -6 in a total of four renal carcinoma cell (RCC) lines prior to and after induction of MDR via exposure to vinblastine. MATERIAL AND METHODS: Surface expression of P-gp and VLA-1 to -6 was determined immunocytochemically in untreated pre-established renal carcinoma cell lines (Caki-1, Caki-2, A498) and a cell line derived from a RCC patient who had received a vinblastine-containing therapy regimen prior to the resection of a local relapse of the tumor (EH). Resistant sublines were cultivated in the presence of 1 ng/ml and 10 ng/ml of vinblastine sulfate, respectively. RESULTS: In all cell lines examined, an increased number of P-gp expressing cells was observed upon exposure to vinblastine. Significant changes of beta 1 integrin expression were observed in three of four RCC cell lines. A de novo expression of VLA-1, VLA-2, and VLA-4 as detected by immunocytochemistry occurred in resistant Caki-1 cells. A498 cells showed an increasing number of VLA-2 positive cells in drug resistant sublines. In contrast, a decrease of VLA-2 and VLA-5 expression was found in EH cells, the only cell line exhibiting P-gp expression prior to vinblastine exposure. Caki-2 cells showed no significant changes of surface integrin expression upon treatment with vinblastine. CONCLUSIONS: Our results demonstrate that induction of drug resistance can be associated with substantial changes of the integrin phenotype in renal carcinoma cell lines. In our experiments, among all VLAs studied, VLA-2 was most frequently altered in expression by RCC cell lines. The significance of these observations for aberrant metastatic properties of multidrug resistant tumor cells will be the subject of further studies. PMID- 10365135 TI - CD44s, E-cadherin and PCNA as markers for progression in renal cell carcinoma. AB - CD44s, E-cadherin and PCNA play an important role in the tumorigenesis of renal cell carcinoma (RCC), although their significance in the clinical progression of RCC is still not clear. In n = 137 cases of operatively resected RCC the expression of CD44s, E-cadherin and PCNA was semiquantitatively analyzed by the APAAP immunohistochemical method. The mean observation period of the n = 74 (54%) of patients with no evidence of disease was 52.6 months and for the n = 63 (46%) patients with progressive disease was 18.7 months. The mean progression free period occurring with absent or low levels of CD44s and PCNA expression was 79 and 118+ months, and with strong expression was 12 and 18 months (p < 0.001), respectively. With strong E-cadherin expression the progression free period was 110+ months, and with low expression 61 months (p = 0.01). A high CD44s/E cadherin ratio and an above average PCNA expression were identified as independent prognostic parameters for the progression tendency of RCC. PMID- 10365136 TI - The differential expression of proinflammatory cytokines IL-6, IL-8 and TNF-alpha in renal cell carcinoma. AB - Recent observations indicate that an antiinflammatory process may play a role in the metastatic cascade of renal cell carcinoma (RCC). Therefore, we compared the expression of cytokines from primary human RCC cultures, from established renal carcinoma cells and those from corresponding proximal renal tubulus cells. For this purpose the different cell types were treated with well defined and with bacterial substances such as the lipopolysaccharide, the staphylococcal enterotoxin B, a superantigen, or a combination of the calcium ionophore A23187 and phorbol 12-myristate-13-acetate. The resulting cell supernatants were analyzed for the proinflammatory cytokines (TNF-alpha, IL-6), the chemotactic active interleukin-8 as well as cytokines from T-helper type I (IL-2, IFN-gamma, IL-12) and type II (IL-4, IL-10). In parallel, the expression of cytokine specific m-RNA was analyzed by multiplex-PCR. Our results clearly demonstrate that among the various cytokines analyzed a predominant release of TNF-alpha, IL 8 and IL-6 is obtained. The remainder cytokines were not detected independent whether molecular biology or cytokine release experiments were applied. Expression of the cytokines was dependent on the degree of malignancy. Among the applied stimuli, only the activation with calcium ionophore/phorbolester modulated cytokine expression and release. While TNF-alpha was induced from normal renal cells by up to 300% (2000 + 120 ng/10(5) cells) a pronounced suppression of TNF-alpha was observed in dependence on the malignancy of the cell line. In contrast, the cytokines IL-6 and IL-8 were significantly upregulated in malignant cells unlike in normal renal cells. These data suggest a differential role of the various cytokines derived from normal or tumor cells. Detailed studies will allow the understanding of the distinct roles of cytokines in renal carcinoma disease. PMID- 10365137 TI - Preclinical studies combining bispecific antibodies with cytokine-stimulated effector cells for immunotherapy of renal cell carcinoma. AB - BACKGROUND: Bispecific antibodies--consisting of a F(ab')-fragment derived from a monoclonal antibody against a tumor epitope as well as of another antibody against a cytotoxic trigger molecule on immune effector cells--can improve the effectiveness of antibody-based tumor therapy. MATERIALS AND METHODS: We used bispecific antibodies with one specifity against the EGF-receptor, which is overexpressed on the majority of renal cell carcinomas, and another specifity against Fc receptors on human leukocytes (Fc gamma RI/CD64; Fc gamma RIII/CD16 and Fc alpha RI/CD89). As source of effector cells, whole blood from patients treated with G-CSF, GM-CSF or IL2/IFN-alpha was used in 51Cr- release assays using various renal cancer cell lines as tumor targets. Further experiments with Percoll-isolated granulocytes or mononuclear cells from the same donors were performed in order to identify the active effector cell populations. RESULTS: Compared with conventional monoclonal EGF-R directed antibodies (murine IgG2a, humanized IgG1), bispecific antibodies induced significantly enhanced cytotoxicity. Highest amounts of tumor cell killing were observed using whole blood from patients treated with G-CSF or GM-CSF in combination with an [Fc alpha RI x EGF-R] bispecific antibody. Under these conditions, granulocytes constituted the most active effector cell population. CONCLUSION: The combination of myeloid growth factors and bispecific antibodies offer a promising new approach for the treatment of advanced renal cell carcinoma. PMID- 10365138 TI - Renal cell carcinoma: immunohistological investigation of expression of the integrin alpha v beta 3. AB - alpha v beta 3 Integrin has been shown in various tumor entities to promote binding to superficial structures of the basement membrane during metastasis. The goal of the present paper was a structural demonstration of this Integrin in renal cell carcinoma (RCC). After removal of paraffin from formalin-fixed tissue, the cells were labelled with the antibody (VNR 147, H. Biermann, Bad Nauheim) using the peroxidase-antiperoxidase method (DAKO Diagnostical GmbH, Hamburg). Evaluation was carried out microscopically and semiquantitatively from missing ( ) through moderate (+) to strong (+2) staining. A total of n = 79 RCCs and n = 53 healthy areas were examined. Semiquantitative staining results: there was a grading-dependent increase (+2) of stainability and thus alpha v beta 3 expression. Two of 53 benign specimens showed strong staining, 11 of 53, only weak staining. Four of 18 G1 RCCs showed strong staining, 11 of 18 only weak staining. Results for G2-RCCs: 11 with strong staining, 21 of 40 with weak staining. G3-RCCs: 4 with strong staining, 2 of 7 with weak staining. Of the metastases, on the other hand, 2 of 14 showed strong staining, another 8 of 14 only weak staining. There were no deviations within the histologic (clear-cell, chromophil, or chromophobe) subpopulations. This grading-dependent expression permits the conclusion that the probability of binding to the human basement membrane mediated by alpha v beta 3 Integrin rises with increasing grading, but the already metastatic cell exhibited this Integrin less strongly, since a basement membrane adhesion is no longer necessary for this cell group. PMID- 10365139 TI - Are renal cell carcinoma cells able to modulate the cytotoxic effect of tumor infiltrating lymphocytes by secretion of interleukin-6? AB - BACKGROUND: Interleukin (IL)-6 can suppress the cisplatin-induced induction of apoptosis in renal cell carcinoma (RCC) in vitro. There are reports on IL-6 secretion by tumor cells. MATERIALS AND METHODS: In supermatants of RCC cultures we measured IL-6 levels by a bioassay. In a videomicroscopic microcytotoxicity assay the immunomodulating effect of secreted cytokines on the cytotoxicity of tumor infiltrating lymphocytes (TIL) was studied. RESULTS: The IL-6 concentrations in 17 supematants of healthy renal cells tended to be higher than in the supernatants of tumor cell cultures. By immunostaining we detected IL-6 in 3/6 cell cultures from healthy renal tissue, in 12/22 tumor tissue samples, and in 8/18 tumor cultures respectively. Supernatants suppressing cytotoxic activity of TIL showed higher IL-6 concentrations in tendency. Suppression of cytotoxicity could also be induced by adding discrete concentrations of IL-6. CONCLUSIONS: IL6 secreted by tumor cells seems to be tumor protecting against cytotoxic TIL. Blocking this mechanism could be an additional approach in immunotherapy of RCC. PMID- 10365140 TI - Renal cell carcinoma: relevance of angiogenetic factors. AB - Angiogenesis is essential for tumour growth. Mostly hypervasculariced renal cell carcinoma (RCC) usually express angiogenic factors, e.g. basic fibroblast factor (bFGF), vascular endothelial growth factor, and angiogenin (ag). This study was designed to evaluate the improvement of serological angiogenesis-factors, their prognostic relevance and correlation to tumour-grade, -stage and -volume. Measurement of bFGF, vEGF and ag was done with ELISA (Quantikine), R and D Systems Europe Abbington, United Kingdom. The control group comprised 39 healthy blood donors, RCC-group were 35 patients with different RCC. Survival dates were presented with Kaplan-Meyer-curves, significance was tested with students-t-test. For all angiogenic factors a highly significant difference between the healthy and RCC-group was found. A strong correlation between angiogenic factors and tumor grade, -stage and -volume, was not found, but a trend for each of the angiogenic factors to correlate with grade was seen. No survival benefit was seen between patients with normal angiogenic factor over those with elevated angiogenic levels. PMID- 10365141 TI - Chemotherapy for renal cell carcinoma. AB - Renal cell carcinoma is a chemotherapy-resistant tumor, which, with commonly used cytotoxic agents, exhibit a only marginal response rate when modern objective response criteria are applied. No clear survival benefit for patients treated with chemotherapy has yet been demonstrated, although most trials indicate that responding patients survive longer. Among the more commonly available cytotoxic agents, vinblastine has been repeatedly reported to achieve a 6-9% objective response rate irrespective of the mode of application. 5-fluorouracil- and Floxuridin (FUDR)-based chemotherapy appear to achieve response rates in the range of 5-8%, with slightly better results when applied by chronomodulation schedules. Trials evaluating the recently licensed newer cytotoxic agents have shown no major improvement for the treatment of metastatic renal cell cancer. Studies including the topoisomerase I inhibitors irinotecan and topotecan are ongoing. The taxanes (paclitaxel, docetaxel) have demonstrated no significant activity and gemcitabine has been reported with responses in single patients. Recent treatment approaches based on the biology of renal cell carcinoma with an increased expression of the multiple drug resistance (MDR) phenotype (pgp 170) have used chemotherapy in combination with verapamil or cyclosporine derivates as inhibitors of the MDR-mechanism. The disappointing results of these trials indicate that MDR is not the only mechanism of chemotherapy-resistance in renal cell carcinoma. In conclusion, chemotherapy has a limited role in metastatic renal cell carcinoma and vinblastine is considered the standard treatment. Strict evaluation of new agents acting independently from the MDR-mechanism is necessary in order to identify drugs with an impact in the palliative treatment of advanced renal cell carcinoma. PMID- 10365142 TI - Adjuvant immunotherapy in renal cell carcinoma--comparison of interferon alpha treatment with an untreated control. AB - BACKGROUND: Adjuvant immunotherapy of RCC with interferon alpha is still controversial. It was the aim of this study to investigate, whether a selected group of patients with a non-metastatic RCC can profit from this therapy. MATERIALS AND METHODS: Survival data of 125 patients with a non-metastatic RCC, who underwent a tumor-nephrectomy were analyzed retrospectively, 33 of these patients received an adjuvant immunotherapy with interferon alpha for 1 year. All tumors were classified by the TNM-system. Statistical evaluation was performed by the Kaplan-Meier-Method and the logrank-test. RESULTS: Tumor stage was seen to be an important prognostic factor in RCC with a significantly better outcome in pT2- as compared to pT3-tumors. Tumor grading was without any prognostic relevance. Adjuvant interferon alpha therapy had no effect on overall survival. After separation of the patients into pT2- and pT3/4-tumors again interferon alpha showed no significant benefit for one of the 2 groups. CONCLUSIONS: The adjuvant therapy with interferon alpha shows no significant benefit in the treatment of non-metastatic RCC. Tumor stage is no suitable factor to select a group of patients, who might profit from an interferon alpha therapy. Further studies are necessary to isolate better selection factors. PMID- 10365143 TI - Treosulfan in the treatment of metastatic renal cell carcinoma. AB - BACKGROUND: Treosulfan is a bifunctional alkylating cytostatic agent that has mainly been used in the therapy of advanced ovarian cancer. Lately, a growth inhibiting effect could be detected in human renal cell carcinoma-cell lines as well. In vitro, Treosulfan showed an even higher growth inhibition than Vinblastine. MATERIALS AND METHODS: We performed a small clinical phase II study using Treosulfan as a monotherapy in the treatment of metastatic renal cell carcinoma. Treosulfan was given to 15 patients with bidimensionally measurable metastases. RESULTS: 10 patients were evaluable. Side effects were negligeable. A complete or even partial remission was not seen. 4 patients showed no change, whereas 6 were progressive. The average time to progress was short (4 months, range 1 to 12 months). CONCLUSIONS: Since Treosulfan did not lead to a measurable tumor remission in the given dose regimen, it does not seem to be suitable for the therapy of metastatic renal cell carcinoma. PMID- 10365144 TI - Life quality of patients with metastatic renal cell carcinoma and chemo immunotherapy--a pilot study. AB - Renal cell cancer (RCC) accounts for 2-3% of all malignant tumors in adults. Due to the indolent course of disease and the few signs and symptoms in early stages the majority of patients presents with metastatic disease when diagnosed. The aims of systemic therapy of RCC are therefore palliative. Recent research shows the key role of immune mechanisms in the course of RCC. The therapeutic use of cytokines, mainly interleukin-2 (IL-2) and interferon-alpha (IFN) results in improvement of remission rates. To date it is unknown to what extent multiple cycles of chemo-immunotherapy alter the life quality (LQ) of patients with metastatic RCC. We monitored life quality during therapy in a three-armed protocol with interferon-alpha 2a, interleukin-2, 5-fluorouracil (5-FU), isotretinoin (ISO) and vinblastin (VBL). Life quality was impaired by two factors: response to chemo-immunotherapy and therapy side effects. A steep decrease of LQ-scores was seen in week 1 of therapy, LQ improved then for patients with stable disease (SD) and partial remission (PR) but not for those with progressive disease (PD). PMID- 10365145 TI - Pathophysiology of tumor angiogenesis and its relevance in renal cell cancer. AB - In many cases, solid tumors exhibit numerous and highly permeable blood vessels. For a long time, this observation drew little attention. In the early 70's, however, Folkman proposed for the first time the potential relevance of tumor angiogenesis (formation of new vessels) for tumor growth and metastasis (6). He realized that tumors up to a diameter of 1-2 mm could be nurtured with oxygen and energy simply by diffusion (prevascular phase). Further growth, however, would require newly formed blood vessels. He hypothesized that (assuming the formation of new vessels was essential for tumor growth) pharmacological inhibition of angiogenesis could be developed as a new, more specific form of tumor treatment. In recent years, several groups have investigated the pathophysiology of tumor angiogenesis. Folkman's hypothesis that tumor growth is dependent on the formation of new vessels was supported by several experiments: Implants of different tumors into an avascular cornea initially have a slow growth rate that increases exponentially after infiltration of new vessels into the tumors (9). Inversely, the growth rate of solid tumors decreases with increasing distance to the supplying capillaries (27). The onset of neovascularization at the bases of human melanomas is directly associated with tumor growth and metastasis (25). Experiments with transgenic mice have demonstrated that the transition from hyperplastic to malignant cell growth occurs parallel to the onset of angiogenesis (7). Tumor vessel density has been shown to be associated with tumor progression and the clinical course in many human tumors (e.g. of the breast, lung, colon, cervix, prostate and bladder). Aside from the basic research on the formation of new (tumor) vessels, it is the therapeutic potential of various inhibitors of angiogenesis, some of which are currently tested in clinical (phase I/II) studies, that deserves special scientific attention. This review gives an overview of the relevance of angiogenesis for tumor growth, especially for renal cancers. It also discusses the potential advantages and disadvantages of different anti-angiogenic therapeutic approaches. PMID- 10365146 TI - Quantification by competitive quantitative RT-PCR of VEGF121 and VEGF165 in renal cell carcinoma. AB - The field of angiogenesis and its mediators in tumour tissue is gaining more and more interest because of their therapeutic implications to arrest progressive growth and metastasis. We examined the expression status of the proangiogenic vascular endothelial growth factor (VEGF) in tumour tissue and adjacent tumour free tissue from renal cell carcinoma (RCC) as well as in cell cultures deriving from tumour tissue. Quantification of both secreted isoforms ot VEGF by competitive RT-PCR revealed a marked increase of VEGF message in tumours and especially in cell cultures from RCC. We found a significant correlation of VEGF expression and microvessel density which was investigated immunohistochemically. As further compared to other urological neoplasms we investigated for VEGF mediated vascularization, RCC is the most promising candidate for anti-angiogenic therapies. PMID- 10365147 TI - Bronchoscopic aspects of renal cell carcinoma (RCC). AB - A symposium on RCC in July 1998 in Tubingen was where we to reviewed our patients with endobronchial metastatic RCC treated by bronchoscopy since 1981. 24 of 26 consecutive cases are the subject of the present study. Tumor history, X-ray findings, endobronchial appearance, treatment and the pathogenetic mechanisms of this special form of metastatic spread are demonstrated and discussed. In 5 patients > 10 years relapsed between nephrectomy and bronchoscopic tumor therapy. A thrombuslike tumor growth and bleeding were striking bronchoscopic features. In X-ray besides atelectasis, hilar or mediastinal masses were seen half of the patients, in 50% without parenchymal lung nodes, suggesting lymphogenic spread. In several patients bronchial recanalisation could be successfully repeated over long periods. Abnormal X-ray, haemoptysis or dyspnoa in patients with RCC demands bronchoscopy. In case of bronchial tumor obstruction bronchoscopic treatment offers excellent palliation. PMID- 10365148 TI - Lymph node involvement in renal cell carcinoma and survival chance by systematic lymphadenectomy. AB - BACKGROUND: The value of systematic lymphadenectomy has been a matter of great controversy for a long period of time. A recently published paper of a retrospective autopsy study generally doubts its therapeutic effectiveness, arguing that positive lymph nodes are nearly always associated with distant metastases. PATIENTS AND METHODS: Between 1974 and 1993 1035 patients suffering from renal cell carcinoma with stages from cT 1 to 4, cM 0 were treated with curative intention. 51% underwent radical abdominal tumour nephrectomy with systematic lymphadenectomy (n = 531, group A). In 199 patients (19%, group B) only macroscopically suspect lymph nodes were removed surgically. All other patients underwent radical lumbar tumour nephrectomy without lymphadenectomy (n = 305, 29%, group C). RESULTS: Mean age of group A was 55.5 +/- 10 years, B 60.3 +/ 11 and C 66.5 +/- 11. Median follow-up for all groups was 115 +/- 63 months. Median amount of removed lymph nodes was 18 in group A, 6 in group B and 3 in group C. N-categories for each group were pN 1: 4%, 2%, 1%; pN 2: 7%, 5%, 1%; pN 3: 3%, 2, %, 1%; pN x: 0%, 35%, 67% respectively. Group A with systematic lymphadenectomy had the least favourable tumour stage overall. Nevertheless long term survival of this group is more favourable with 57% +/- 6 when compared to group B with 50 +/- 12% and C with 44% +/- 9%. 20 (27%) of the 75 lymph node positive patients of group A who have been followed-up for more than 5 years are still alive. CONCLUSIONS: At least 4% of all patients benefit from extensive lymphadenectomy. This may only be a relatively small proven effect for the entire patient collective, but for a single lymph node positive patient this is an undoubtedly significant additional chance of survival especially when one notes that presently there is no curative adjuvant therapy. PMID- 10365149 TI - Brain metastasis in renal cell carcinoma: clinical data and neuropathological differential diagnoses. AB - Beside lung and skeleton, renal cell carcinoma (RCC) can also metastasize to the brain. In order to collect clinical and histopathological data on these tumors, we studied the local distribution of 50 metastases of RCC in different brain regions (frontal, parietal, temporal occipital lobe and brain stem/cerebellum) and found frequencies ranging from 18.4% to 22.4% in each region. This indicates that there is no preferable site of renal cell carcinoma metastases in the brain. In 46 (92%) of the cases the primary tumor or metastases in other organs had already been diagnosed before operation of a brain metastasis. This shows that the latter mostly is a late manifestation at an advanced stage of disease. 44 (88%) of the metastases showed the typical clear cell pattern. In case of unknown primary tumor, these have to be distinguished from other potentially intracranial neoplasms with similar histologic features. These are chordoma, germinoma, paraganglioma, alveolar sarcoma of soft tissue and the epitheloid variant of malignant peripheral nerve sheath tumor. PMID- 10365150 TI - Possible synergy of radiotherapy and chemo-immunotherapy in metastatic renal cell carcinoma (RCC). AB - PURPOSE: Bone metastases or local recurrences are widely viewed as poor prognostic signs for successful immunotherapy for metastatic renal cell carcinoma (RCC), and even partial remission is a rarity. We assessed the efficacy of the combination of radio and chemo-immunotherapy for bone metastases or local recurrences form RCC. MATERIALS AND METHODS: From February 1994 until September 1997 twelve patients with progressive renal cell carcinoma (9 with bone metastases and 3 with local recurrence) were treated with a combination of chemo immunotherapy and radio therapy. RESULTS: Four out of twelve patients achieved complete remission (CR), one patient had a partial remission (PR), three were stable and four had disease progression under radio therapy and chemo immunotherapy. Yet three pts. died of the disease. The toxicity symptoms according to WHO ranged between grade 2 and grade 3. CONCLUSION: Our data suggest that the combination of radio therapy and chemo-immunotherapy may induce a synergistic antitumor effect for the treatment of bone metastases or local recurrences from renal cell carcinoma. PMID- 10365152 TI - Resection of pulmonary metastases from renal cell carcinoma. AB - Between 1980 and 1995, 77 patients underwent complete resection of pulmonary metastases from a renal cell carcinoma after exclusion of a primary tumor recurrence and other metastatic localizations. 30-day mortality was 3%. The Median follow-up was 34 months (M). Cumulative 5-year survival (5-YS) was 39%. Prognostic criteria are the duration of the disease-free interval (DFI) and the number of metastases. Patients with a DFI > or = 48 M had a 5-YS of 46% compared to 26% for a DFI of < 48 M. Patients with a solitary metastasis had a 5-YS of 49% compared to 19% for multiple metastases. There was no significant difference in terms of sex, kind of access, kind of operation, and unilateral or bilateral affection. Since metastases from renal cell carcinomas are almost resistant to chemotherapy and radiotherapy and immunotherapy at present does not considerably improve long-term survival, surgical resection currently is the only effective therapeutic access in renal cell cancer metastasized to the lung. PMID- 10365151 TI - Matrix metalloproteinases and their inhibitors. AB - Degradation of the extracellular matrix around tumour cells is an essential step in the process of tumour invasion and metastasis. The family of structurally related metalloproteinases (MMPs) and their inhibitors, the tissue inhibitors of metalloproteinases (TIMP), play an important role in matrix degradation by tumour cells. In this paper MMP and TIMP activity was analysed in renal cell carcinoma. On the transcriptional level, RT-PCR was used to evaluate the expression of membrane type (MT) 1-MMP, MMp-2, MMP-9 and the inhibitors TIMP-1 and TIMP-2 in tumour tissue and normal kidney tissue. Zymography and reverse zymorgaphy measured the activity of MMPs and TIMPs in the supernatant of primary cell cultures derived from these tumour tissues at the protein level. In addition, MMP and TIMP serum levels of these patients were analysed before and 7 days after tumour nephrectomy. MMP expression revealed to be five times higher in low graded tumour tissue compared to normal kidney tissue. In the supernatant of the cell cultures, both MMP-2 and TIMP protein level was higher in samples derived from advanced carcinoma compared to samples derived from organ confined tumours. Serum levels of TIMP-2 decreased significantly after tumour nephrectomy. In conclusion, MMPs are key enzymes for tumour progression in renal cell carcinoma. New functions especially concerning the TIMP proteins may provide further understanding of the tumour progression processes. PMID- 10365153 TI - Perioperative brachytherapy as an additional therapeutic option in patients with renal cell carcinoma (RCC), either inoperable or after completed percutaneous radiotherapy. AB - Between January 1992 and December 1997 60 patients with different tumors were treated with perioperative high dose rate brachytherapy with Iridium 192 in afterloading technique (HDR), total dosis 20-30 Gy. Three of the patients had RCC. These patients are presented in detail. In elected patients in whom other forms of treatment are not available or have failed, the method offers an additional therapeutic option and can achieve excellent results. PMID- 10365154 TI - Future strategies in external radiation therapy of renal cell carcinoma. AB - The advantage of external radiation therapy in renal cell carcinoma is controversial. High complication rates reported in previous trials of postoperative radiation therapy can now be avoided by using contemporary modern treatment techniques. Based on both prospective and retrospective data the following indications for adjuvant radiation therapy should be considered and tested in phase III trials: a) unresectable non-metastatic tumours (preoperative irradiation), b) incomplete resection with gross or macroscopically positive margins, c) locally advanced tumour with perinephric fat extension or adrenal invasion. PMID- 10365155 TI - New treatment modalities--the urologist's view. AB - Therapy of metastatic RCC is still unsatisfactory. To date, biochemotherapy with IL-2 s.c., IF a2 s.c. and 5-FU i.v. is probably the best treatment available. Response rates range between 30 and 40%. Toxicity is tolerable. Patients belonging to the low risk group, (presenting none of the following risk parameters: BSR > 70 mm, LDH > 280 U/l, neutrophilic granulocytes > 6000/microliter, hemoglobin < 10 g/l, extrapulmonary or bone metastases) show the best results. Response rates of 60% and 2-year survival rates of 65% can be achieved. When selecting candidates for biotherapy, risk parameters as mentioned above should be considered. Patients presenting these criteria do not profit from biochemotherapy. Nephrectomy in metastatic RCC is indicated in symptomatic patients. As part of an integrated treatment regimen with biotherapy in asymptomatic patients, nephrectomy should be performed only after response to biotherapy. These patients show a long-lasting progression-free survival. A randomized study proving the benefit of cytoreductive surgery, however, does not exist. Experimental research recently developed numerous strategies for potential therapy. The most interesting and hopeful starting points are gene manipulation and angiogenesis. Clinical trials, however, have to be awaited. PMID- 10365156 TI - Radiosurgery for the treatment of brain metastases in renal cell carcinoma. AB - BACKGROUND: In the treatment of brain metastases using a stereotactically modified linear accelerator it could be shown that a single dose between 15 and 25 Gy leads to partial or complete remission of so-called radioresistant metastases from melanoma and hypernephroma. Radiosurgery of brain metastases then started in centers all over the world, however, experiences with brain metastases of renal cell carcinoma are yet limited. The aim of this analysis is therefore to present the treatment results of radiosurgery of brain metastases. Furthermore, in this paper prognostic subgroups shall be defined, in order to establish guidelines for an optimal therapy strategy. MATERIALS AND METHODS: Radiosurgery means stereotactically guided high-precision irradiation methods by extremely focussing ionizing radiation within the target volume as a single dose application. The characteristic steep dose decrease allows the selective destruction of small intracranial lesions, while the surrounding brain tissue is optimally protected. Two methods, Gamma Knife and stereotactic modified linear accelerator are clinically available. RESULTS: In larger studies from different groups all over the world, local tumor control rates from 85% to 95%, recurrence rates from 6% to 15% and side effects between 3% and 15% have been attained, independent of the system used. Prognostic factors, like volume of metastases < 10 ml, applied dose > 18 Gy, one or two metastases, absence of extracranial metastases, good patient performance with a Karnofsky score > 70%, primary treatment and more than one year between primary diagnosis and brain metastases showed a trend toward improved survival. Depending on the prognostic factors the median survival after radiosurgery ranged from 6 to 12 months. Retrospective comparison of radiosurgery and surgical series suggest that both modalities attain similar results. The dose can be applied with an accuracy of 0.3 mm. DISCUSSION: Based on these experiences, brain metastases can be treated by radiosurgery, primarily in patients with one or two metastases or in combination with whole brain irradiation as a boost in patients with more than two metastases. Furthermore with radiosurgery a new treatment modality exists to re irradiate patients who have been failed after surgery or whole brain irradiation. PMID- 10365157 TI - Metastatic spine disease in renal cell carcinoma--indication and results of surgery. AB - Metastatic spine disease is frequent in renal cell carcinoma and 50% of osseous metastases are already found at the time of primary diagnosis. Therefore patient mobility and quality of life are threatened early in the course of disease. Surgery is able to relieve pain and to regain or to preserve mobility. Indication and technique of surgery (anterior decompression, vertebral replacement and transpedicular fixation) are explained and treatment results of eleven cases are reported. All patients with paraparesis or cord compression preoperatively were mobile when leaving our hospital after surgery. There were no severe complications, especially no neurological deteriorations or deaths. Postoperative survival time was ten months approximately in cases with multiple osseous lesions and it was several years in cases with solitary metastases. Mobility was preserved for most of the survival time. In conclusion, restabilisation of the spine proved to be a worthwhile treatment option in well-selected cases suffering from malignant spinal involvement. PMID- 10365158 TI - Evolution of herpes simplex virus type 1 under herpesviral evolutionary processes. AB - Herpesviruses, the genomes of which are double-stranded DNA of 120 kilobase pairs or more, infect a wide range of vertebrates from mammals to fish. Herpes simplex virus type 1 (HSV-1), a representative of family Herpesviridae, is a ubiquitous human pathogen. HSV-1 relates to common mucocutaneous diseases, while HSV-1 infection can mean a serious outcome, e.g. blindness and insult to the central nervous system. Evolution of herpesviruses includes DNA rearrangements, often generating tandemly or invertedly repeated sequences. Studies of HSV-1 DNA dynamics substantiated these processes of DNA recombination involved in the evolution of herpesvirus. Herpesviruses seem to have diversified from a common ancestor, in a manner mediating co-speciation of herpesviruses with host species through species-specific latent infections. Thus, the notion of host-linked evolution of herpesviruses is given support. Relationships between HSV-1 genotypes and human ethnic groups can be traced by analyses of DNA polymorphisms of HSV-1 strains present in populations of various countries. A close association of an HSV-1 genotype with a particular historical human population seems probable. Such being the case, the host-linked mode is likely to be linked to diversification of HSV-1 in human populations. PMID- 10365159 TI - Viral coinfection in salmonids: infectious pancreatic necrosis virus interferes with infectious hematopoietic necrosis virus. AB - Coinfection of farm-reared salmonids involving two viruses has been described, but there is no report on the interactions between viruses. Here we examine whether infectious pancreatic necrosis virus (IPNV) strain Sp interferes with the growth of infectious hematopoietic necrosis virus (IHNV) strain S46, a coinfected isolate from rainbow trout. When BF-2 cell culture was inoculated with S46 the infective titer of the IHNV fraction decreased by 3 log10 units compared to the growth curve of IHNV in the single infection. RT-PCR assay confirmed this reduction, which after successive passages of the co-infected sample led to a decrease in IHNV mRNA and the absence of the specific PCR product for IHNV. Flow cytometry showed that only 13% of the cells inoculated with S46 strain were infected with IHNV at 48-72 h post infection, in contrast to the 50-80% of cells that were positive for IPNV. Exposure of cells to IHNV for 24 h before infection with IPNV did not affect the infective titers of either virus or the PCR results obtained in simultaneous coinfections. Moreover IHNV was not inhibited when the IPNV inoculum was reduced. So, a multiplicity of infection dependence was demonstrated for IPNV-IHNV interference; the RT-PCR assay described here was found to be a suitable technique for identifying and studying dual viral infections. PMID- 10365160 TI - Exacerbation of influenzavirus pneumonia by intranasal administration of surfactant in a mouse model. AB - Although surfactant-secreting type II alveolar cells have been shown to be damaged during influenzavirus pneumonia, little is known about the effects of surfactant replacement therapy. We have developed a mouse influenza model, in which viral infection can be localized to the upper respiratory tract or to both the upper and lower respiratory tract depending on the volume (rather than infectious dose) of intranasal inocula of influenzavirus. In this model, only mice infected with a large inocula die with massive infection in the lung. Using this model, we unexpectedly found that intranasal administration of surfactant dramatically exacerbated influenzavirus infection causing fatal disease even in mice inoculated with a small inocula. This exacerbation resulted from enhancement of intrabronchial and intraalveolar spread of virus, as confirmed by immunohistochemical detection of viral antigen in lungs. Assuming this experimental model in mice recapitulates naturally occurring disease in humans, extreme caution is warranted in surfactant-replacement treatment of influenzavirus pneumonia in humans. PMID- 10365161 TI - Characterisation of non-structural protein 3 of hepatitis C virus as modulator of protein phosphorylation mediated by PKA and PKC: evidences for action on the level of substrate and enzyme. AB - Generally, the maximum activities of the protein kinases A (PKA) and C (PKC) show an optimum value for their substrate concentrations rather than a saturation curve; at high substrate concentrations, the kinase activity is completely abolished. The C- and N-truncated form of the non-structural protein 3 (NS3) of hepatitis C virus (HCV) (HCV-polyprotein-(1,189-1,525)) abolishes the inhibiting effect of the substrate, yielding saturable Michaelis-Menten kinetics of PKA and its catalytical domain (C subunit). In contrast, HCV-polyprotein-(1,189-1,525) activates PKC with increasing Vmax, while it abolishes the substrate inhibition of its catalytical domain (M-kinase) through a mechanism analogous to that of PKA and C subunit. PKC isoforms alpha, beta and gamma investigated are similarly activated by HCV-polyprotein-(1,189-1,525). Our data suggest that NS3 attenuates the substrate inhibition through a generalized mechanism operating mainly on the substrate level that directly results from a specific protein-protein interaction. In the case of the PKC, an additional kinase activating mechanism operates on the enzyme level. Both actions of NS3, the attenuation of the substrate inhibition and the activation of PKC, could not be explained by classical means that predict autophosphorylation to enhance the rate of substrate phosphorylation. The results are discussed in view of similar activities displayed by matchmakers and some molecular chaperones. PMID- 10365162 TI - Characterization of tobacco geminiviruses in the Old and New World. AB - Biological differences and molecular variability between six phenotypically distinct tobacco-infecting geminivirus isolates from southern Africa (Zimbabwe) and Mexico were investigated. Host range studies conducted with tobacco virus isolates ZIM H from Zimbabwe and MEX 15 and MEX 32 from Mexico indicated all had narrow host ranges restricted to the Solanaceae. Alignment of coat protein gene (CP) and common region (CR) sequences obtained by PCR, and phylogenetic analysis of the CP sequences indicated Zimbabwean isolates were distantly related to those from Mexico and that geographically proximal isolates shared their closest affinities with Old and New World geminiviruses, respectively. Zimbabwean isolates formed a distinct cluster of closely related variants (> 98% sequence identity) of the same species, while MEX 15 segregated independently from MEX 32, the former constituting a distinct species among New World geminiviruses, and the latter being a variant, Texas pepper virus-Chiapas isolate (TPV-CPS) with 95% sequence identity to TPV-TAM. Results collectively indicated a geographic basis for phylogenetic relationships rather than a specific affiliation with tobacco as a natural host. MEX 15 is provisionally described as a new begomovirus, tobacco apical stunt virus, TbASV, whose closest CP relative is cabbage leaf curl virus, and ZIM isolates are provisionally designated as tobacco leaf curl virus, TbLCV ZIM, a new Eastern Hemisphere begomovirus, which has as its closest relative, chayote mosaic virus from Nigeria. PMID- 10365163 TI - Pathogenic, antigenic and molecular properties of rabbit haemorrhagic disease virus (RHDV) isolated from vaccinated rabbits: detection and characterization of antigenic variants. AB - Rabbit haemorrhagic disease virus (RHDV) isolates were obtained from several animals previously vaccinated with an inactivated vaccine. Seven isolates were analyzed by immunological and molecular biological methods and compared to reference strains. Antigenic characterization with monoclonal antibodies as well as haemagglutination assays demonstrated considerable differences between individual isolates. However, sequencing of the capsid protein genes revealed a high degree of homology between five of these isolates and the reference strain FRG. In contrast, two isolates specified remarkably different capsid proteins with a degree of variation not observed so far in RHDV. Amino acid alterations were found clustered between residues 301 and 328 (region C), 344 and 434 (region E) and also in the 3' region of the capsid protein gene. Interestingly, experimental vaccination of rabbits followed by challenge with the heterologous variant strains showed restricted cross-protection against one of the strains. In summary, we found a level of antigenic variation not detected in RHDV so far, and describe two distinct new antigenic variants. PMID- 10365164 TI - Choristoneura fumiferana granulovirus: sequence analysis and 5' characterization of ORF891. AB - A gene located immediately upstream of the granulin gene of Choristoneura fumiferana (ChfuGV) granulovirus was identified, sequenced and named ORF891. The determined, putative open reading frame (ORF) of 891 bp encodes an estimated 34.6 kDa protein. The 5' end transcript of the gene was mapped and analysed. A putative promoter region organization of ChfuGV ORF891 contains a consensus late baculovirus promoter element, TAAG, and two putative early TATA boxes similar to the promoters of ORF909 of Cryptophlebia leucotreta granulovirus (ClGV). Sequence comparisons of ChfuGV ORF891 with ClGV ORF909 and Cydia pomonella granulovirus (CpGV) ORF124R showed respective homologies of 60.9 and 63.9% for nucleotides and 46.3% and 49.3% for amino acids. Homology of ChfuGV ORF891 with ME53 ORF of Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) was 68.2% for nucleotides but a total lack of homology for amino acid sequences. Two zinc finger motifs are also associated with ChfuGV ORF891. PMID- 10365165 TI - The barriers to bluetongue virus infection, dissemination and transmission in the vector, Culicoides variipennis (Diptera: Ceratopogonidae). AB - Transmission of bluetongue virus (BTV) by a vector species of Culicoides was studied using immunohistochemistry, virus titration and in vitro transmission tests. Adult female C. variipennis were used from two colonies that are either "transmission competent" or "transmission refractory" after oral infection with BTV. Intrathoracic (i.t.) injection of BTV into the haemocoel always resulted in a fully disseminated infection and transmission of virus in saliva. However, after ingestion of an infectious blood meal, only 30% (approximately) of midges from either colony became persistently infected. Although none of the orally infected insects from the "refractory" colony were able to transmit virus, 12% of those from the "competent" colony (containing > or = 10(3.0)TCID50 of virus/midge) did transmit BTV in their saliva. The most important barriers to BTV transmission in Culicoides vector species appeared to be a mesenteron infection barrier (MIB), which controls initial establishment of persistent infection, a mesenteron escape barrier (MEB) which can restrict virus to gut cells and a dissemination barrier (DB) which can prevent virus which enters the haemocoel from infecting secondary target organs. Culicoides variipennis do not appear to present either a salivary gland infection barrier (SGIB), or a salivary gland escape barrier (SGEB) to BTV. PMID- 10365166 TI - Characterisation of a recent virulent transmissible gastroenteritis virus from Britain with a deleted ORF 3a. AB - Analyses of transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus (PRCV) isolates have suggested that tropism and pathogenicity are influenced by the spike protein and ORF 3. In general, enteric viruses (TGEV) have been shown to contain intact spike and ORF 3 genes, whilst respiratory isolates (PRCV) have major deletions within both regions. Virulence has been correlated to a functional ORF 3. Here, sequence analysis of a recent isolate of virulent TGEV, revealed a variant with an intact spike gene, but a large deletion in ORF 3a. This suggests that ORF 3a is not essential for enteric virulence. PMID- 10365167 TI - Detection of equine herpesvirus types 2 and 5 (EHV-2 and EHV-5) in Przewalski's wild horses. AB - In blood samples of seven captive equid species from four German zoos EHV-1 specific antibodies were detected in 76% and EHV-4 specific antibodies in 73% of the 55 animals, whereas 93% were tested positive for EHV-2 and EHV-5, respectively. In only one blood sample from a Przewalski's wild horse EHV-4 DNA was amplified by PCR. From seven Przewalski's wild horses EHV-2, and from another one EHV-5 was isolated by cocultivation. The identity of the virus isolates was verified by PCR and restriction enzyme digestion. PMID- 10365168 TI - Strain-specific differences in the effect of influenza A virus neuraminidase on vector-expressed hemagglutinin. AB - In order to evaluate the efficiency of the removal of sialic acid residues from the influenza virus hemagglutinin by the viral neuraminidase in the course of the virus replication cycle, CV-1 cells expressing the hemagglutinin of H7 subtype from an SV40-based vector were superinfected with influenza virus strain A/Duck/Ukraine/63 (H3N8) or A/USSR/90/77 (H1N1). Vector-expressed hemagglutinin was immunoprecipitated from cell lysates and analyzed by polyacrylamide gel electrophoresis. The data indicate that the removal of sialic acid residues from the vector-expressed H7 hemagglutinin by N1 neuraminidase of A/USSR/90/77 virus in the course of the virus replication cycle is incomplete. The results are discussed in connection with previously published data showing that the low activity of NA in wild-type influenza virus results in incomplete removal of sialic acid residues from virion components. PMID- 10365170 TI - In vitro evidence for RNA binding properties of the coat protein of prunus necrotic ringspot ilarvirus and their comparison to related and unrelated viruses. AB - The RNA binding properties of the prunus necrotic ringspot virus (PNRSV) coat protein (CP) were demonstrated by northwestern and dot-blot analyses. The capability to bind PNRSV RNA 4 was compared with viruses representing three different interactions prevailing in the assembly and architecture of virions. The results showed that cucumber mosaic virus (CMV) and PNRSV CPs, which stabilise their virions mainly through RNA-protein interactions bound PNRSV RNA 4 even at very high salt concentrations. The CP of cherry leaf roll nepovirus, whose virions are predominantly stabilised by protein-protein interactions did not bind even at the lowest salt concentration tested. Finally the CP of carnation mottle carmovirus, that has an intermediate position in which both RNA protein and protein-protein interactions are equally important showed a salt dependent RNA binding. PMID- 10365169 TI - Adenovirus cellular receptor site recirculation of HeLa cells upon receptor mediated endocytosis is not low pH-dependent. AB - HeLa cells were depleted of 75% of the available adenovirus type 2 specific cellular receptor sites (CRSs) by controlled attachment of viruses at a high multiplicity of infection. Upon removal of unattached viruses and further incubation, the cells were able to recycle the CRSs to 75% of the level of uninfected control cells within 5 h. The rate of virus receptor recycling was approx. 1 CRS x min-1 x cell-1. The existence of receptor recycling further strengthens the involvement of a total process of receptor-mediated endocytosis in adenovirus internalization. The recirculation process was neither affected by the lysosomotropic agents ammonium chloride and amantadine-HCl, nor by the ionophore monensin or the multifunctional weak-base amine chloroquine. PMID- 10365171 TI - First detection of bovine group B rotavirus in Japan and sequence of its VP7 gene. AB - An epizootic outbreak of diarrhea occurred in adult cows on a dairy farm in Hokkaido, Japan. One colostrum-fed calf inoculated with pooled feces of the 5 affected cows, developed mild diarrhea, and shed rotavirus-like particles which reacted with antiserum to group B rotavirus in immune electron microscopy. Cell culture immunofluorescence tests, RNA-polyacrylamide gel electrophoresis and RT PCR confirmed that this virus was bovine group B rotavirus, which was designated the Nemuro strain. Additional 2 colostrum-deprived calves inoculated with feces of the first calf also developed diarrhea and shed virus, suggesting that this group B rotavirus might be the etiological agent of the outbreak of adult cow diarrhea. The identities of the nucleotide (nt) and deduced amino acid (aa) sequences of the Nemuro VP7 gene were high (93-95% in nt and 96-97% in aa) and low (61-63% in nt and 49-61% in aa) compared to those of the published corresponding genes from 3 bovine and 2 other mammalian (human and rat) strains of group B rotaviruses, respectively. To our knowledge, this is the first report on the presence of bovine group B rotavirus in Japan. PMID- 10365172 TI - Phylogenetic characterization of a highly attenuated strain of equine arteritis virus from the semen of a persistently infected standardbred stallion. AB - An avirulent, novel variant of equine arteritis virus (EAV; CA95G) was isolated from the semen of a persistently infected Standardbred stallion. The CA95G virus caused subclinical infection and seroconversion in susceptible horses, and virus was isolated only once from blood and nasal secretions collected from 6 experimentally infected horses. Sequence analysis of genes encoding the known EAV structural proteins shows that this highly attenuated strain of EAV is genetically similar to virulent field strains of EAV and, in particular, to a strain of EAV that was isolated during an outbreak of equine viral arteritis in western Canada in 1986. Not only is the carrier stallion the critical natural reservoir of EAV, but genetic diversity of the virus is generated in the course of persistent infection of carrier stallions. The subtle genetic changes that facilitate and maintain persistent EAV infection of the stallion's reproductive tract likely influence phenotypic properties of the virus such as virulence. PMID- 10365173 TI - PrP genotype frequencies of the most dominant sheep breed in a country free from scrapie. AB - Polymorphisms within the prion protein (PrP) gene are associated with scrapie susceptibility. We analysed the PrP genes of 140 Romney Marsh sheep, the dominant breed in New Zealand, a country free from scrapie. We found PrP alleles that are associated with a high susceptibility to scrapie. Sheep with these PrP genotypes would probably succumb to scrapie when born and raised in a scrapie endemic environment. These findings correspond to those obtained in minor breeds from New Zealand. We conclude that scrapie development not only depends on host genetic factors but also requires exogenous factors. Our findings demonstrate the effectiveness of the measures taken by New Zealand to maintain free from scrapie. PMID- 10365174 TI - Progress and controversy in Bornavirus research: a meeting report. PMID- 10365175 TI - [The effect of ABO, Rhesus and Kell blood group antigens on gallstone prevalence. A sonographic study of 1030 blood donors]. AB - BACKGROUND AND OBJECTIVE: Besides generally accepted risk factors of the pathogenesis of gallstone disease such as age, obesity, female sex and high number of births, hereditary factors are held responsible for different prevalence rates. A number of studies dealt with the question of a correlation between the prevalence of gallstone disease and different blood groups. The Ulm Gall Bladder Stone Study represents the first sonographic prospective study regarding this issue. SUBJECTS AND METHODS: Unselected blood donors (n = 1030, 606 men, mean age 38.0 years, 424 women, mean age 34.1 years) were sonographically examined for presence of gallstones at the German Red Cross blood donor centre in Ulm. Besides AB0, Rhesus and Kell blood group anthropometric data of the test subjects were recorded by means of a semi-standardized interview. RESULTS: The prevalence of gallstone disease in all test subjects was 6.0%. Within the AB0 system the prevalence in subjects with blood group AB was highest (12.1%). The prevalence in Rh-positive and Rh-negative subjects was nearly identical (6.0 vs. 6.1%). Kell factor positive subjects suffered less from gallstone disease than Kell factor negative subjects (2.0 vs. 6.3%). None of these differences in prevalence were statistically significant. CONCLUSION: This study revealed no significant correlation between the distribution of the AB0, Rhesus and Kell blood group antigens and the prevalence of gallstone disease. PMID- 10365176 TI - [Diffuse panbronchiolitis. A rare differential diagnosis of chronic obstructive lung disease]. AB - HISTORY AND ADMISSION FINDINGS: A 36-year-old Korean had since childhood suffered from a chronic progressive lung disease, marked by dyspnoea, productive cough and recurrent sinusitis. He was in a reduced general condition on admission. There were hyperresonant sounds on palpation over both lungs, a low diaphragm and slight vesicular breath sound as well as occasional rales and sibilant rhonchi. INVESTIGATIONS: Laboratory tests showed increased blood cell sedimentation of 52 mm at 1 h, leukocytosis of 24.3/nl, and a raised cold-agglutinin titre. Haemophilus influenza and Pseudomonas aerogenes were cultured from the bronchial secretion. Lung function tests revealed severe obstruction with hyperinflation and partial respiratory failure with a much reduced 1-sec forced expiratory volume (FEV1) of 0.71, 19% of normal; residual volume (RV) 6.61, 550% of normal; arterial oxygen partial pressure 61 mm Hg; arterial CO2 partial pressure 36 mm Hg. Chest X-ray showed emphysematous changes with diffuse small nodular shadows. Bronchoalveolar lavage revealed 96% neutrophils, indicating marked granulocytic inflammation. A video-assisted thoracoscopic biopsy was performed which showed diffuse panbronchiolitis. TREATMENT AND COURSE: The symptoms, radiological changes and lung functions clearly improved under long-term treatment with 500 mg erythromycin daily. CONCLUSION: Diffuse bronchiolitis is rarely seen outside of Asia, but it should be included in the differential diagnosis because, in contrast to other forms of chronic obstructive lung disease, it responds to long term erythromycin medication. PMID- 10365177 TI - [Tinea capitis and corporis caused by Trichophyton soudanense in an immigrant family from Africa]. AB - HISTORY AND FINDINGS: Several weeks before coming to Germany the two daughters (aged 3 and 6 years) of a family from Togo had developed desquamating skin changes over the hairy scalp. These had then spread to the trunk and limbs. The 8 weeks-old son also had discrete lesions on the hairy scalp and neck. In all of them these lesions had then spread and begun to itch markedly. When first seen as out-patients the father was free of symptoms, but the other members of the family had multiple, sharply circumscribed, partly confluent, dry and desquamating lesions, about 2-4 cm in diameter, with areas of alopecia and hair breaking off at skin level. In addition there were dry, desquamating, sharply circumscribed, partly hyperpigmented, partly infiltrated plaques, 1-3 cm in diameter, disseminated over the entire body surface, but especially the neck and limbs. INVESTIGATIONS: Typical micromorphological characteristics for T. soudanese were demonstrated in the outer zones of a primary culture and the organism was also demonstrated in culture on Sabouraud-glucose-agar. Typical colonies on Lowenstein Jensen medium allowed differentiation from Microsporum ferrugineum. TREATMENT AND COURSE: The patients were treated systemically with griseofulvin and locally with ciclopiroxolamine. Marked clinical improvement occurred within 2 months and cultures became negative. But as fungal elements were still demonstrated in native preparations from two of the patients, treatment was continued. CONCLUSION: Efficacious treatment of tinea needs reliable diagnosis of the pathogen. Human infection with T. soudanese usually results from contact with other humans. If this infection occurs in persons not from Africa there is usually the history of indirect or direct contact with Africans. Increased international migration and tourism is likely to result in more cases of this kind: this pathogen should be considered in the differential diagnosis of tinea of scalp and body. PMID- 10365178 TI - [Therapy of multiple sclerosis]. PMID- 10365179 TI - [Polymorphisms--genetic risk factors for coronary heart disease?]. PMID- 10365180 TI - [Magnetic resonance tomography for eclampsia during pregnancy?]. PMID- 10365182 TI - 'Handbook for mortals'. PMID- 10365181 TI - [Massive duodenal lambliasis and Helicobacter heilmannii gastritis in a diabetic patient]. PMID- 10365183 TI - Geriatrics photo quiz. The physician. Colles' fracture. PMID- 10365184 TI - Carotid occlusive disease: primary care of patients with or without symptoms. AB - Of the half-million strokes that occur each year in the United States, 20 to 30% can be directly linked to carotid occlusive disease. The degree of stenosis involving the carotid bifurcation is an important predictor of stroke risk. Asymptomatic disease may be diagnosed on routine physical exam or screening of the carotid bifurcation in patients with risk factors for ischemic strokes. Symptomatic disease includes transient ischemic attacks, stroke in evolution, and complete stroke. Duplex ultrasound scanning is the standard test for the initial evaluation of carotid artery disease. Patients undergoing surgery should also have magnetic resonance angiography or an angiogram of the carotid vessels. Stroke prevention includes lifestyle modification such as cessation of smoking, strict dietary and medical management of hyperlipidemia, diabetes, and hypertension. Antiplatelet, anticoagulant, and thrombolytic therapy can be used where indicated. PMID- 10365185 TI - Hypertension: therapeutic approach to weight loss, exercise, and salt intake. AB - Ideal body weight is difficult to achieve, but losing 10 lbs may be beneficial in controlling high blood pressure. Patients are more likely to exercise if they enjoy the activity, establish a routine, and exercise with a friend. Older Americans consume a lot of processed foods, the source of 80% of dietary salt. Most older African-Americans, especially those with obesity and type 2 diabetes, are salt-sensitive. Four steps to successful patient education are: 1) tell patients their blood pressure readings; 2) tell them about their medications and potential side effects; 3) provide culturally-sensitive printed materials; 4) use video tapes to educate all new patients and those with compliance problems. PMID- 10365186 TI - Late-life depression: how to treat patients with comorbid chronic illness. Interview by Alice V. Luddington. AB - In persons age 65 and older, the incidence of depression increases with the degree of physical health problems. Higher levels of mortality among depressed patients may be attributed to psychological stress, which triggers the production of cortisol by the adrenal glands and thereby adversely affects the immune system. Some 70 to 90% of late-life depression is undiagnosed; this often occurs if the patient's depressive symptoms could be attributed to other medical problems. Screening for depression can be done in the primary care office in about 1 minute. Older patients with mild depression may need no more than a counselor with good listening skills. Moderate to severe depression may require antidepressant therapy, usually with very low initial doses. An epidemic of depression that is expected in the next century will require physicians to utilize community resources to care for the aging 'baby-boom' generation. PMID- 10365187 TI - Lichenified plaques. Scratching exacerbates the pruritic eruption and causes crusted skin markings. PMID- 10365188 TI - Identifying prenatal alcohol use: screening instruments versus clinical predictors. AB - The purpose of this study is to compare the accuracy of screening instruments with clinical predictors in the identification of prenatal alcohol use. 350 women initiating prenatal care at the Brigham and Women's Hospital (Boston, MA) completed the T-ACE, AUDIT, and SMAST. The predictive accuracy of each was compared using Receiver Operating Characteristic (ROC) curve analysis. The T-ACE, AUDIT, and clinical predictors alone correctly identified 65 to 70% of current drinkers, whereas the SMAST alone performed only slightly better than chance. The predictive ability of the T-ACE was further improved with the addition of clinical predictors. PMID- 10365189 TI - Predictors of change in frequency of crack cocaine use in a street-recruited sample. AB - Crack cocaine users are at high risk for HIV, with higher frequency crack users engaging in higher rates of HIV-related sexual risk behaviors. This study will assess the variables impacting changes in crack use frequency. Out-of-treatment crack users were street recruited in East Harlem, NY. Subjects (n = 727) were 33% female, 91% minority, and 28% reported recent drug injecting. Baseline and 6 month follow-up interviews were administered. There was a significant reduction in crack use over time (p < .0001). Subjects were categorized according to five groups, based on their change in level of crack use between the two interviews, to predict those who stopped, maintained, or changed their level of use. Discriminant analyses identified six variables as the best predictors of the five groups, including having been in drug treatment since baseline and having been a drug injector (both related to reduced levels of crack use). The overall reduction in crack use for the sample masked the fact that important subgroups remained at high use levels or increased their use. The identification of subgroups who may be most resistant to reducing drug use can be helpful in developing more effective interventions. PMID- 10365190 TI - Personality dimensions and substance misuse: relationships in adolescents, mothers and fathers. AB - The research addressed the question of whether relationships exist between personality dimensions, antisocial behavior, and alcohol or other substance misuse (AOSM) in adolescents and in their fathers and mothers, who often also have histories of AOSM. One hundred male adolescents (mean age 15.8 years) entering a residential treatment center for youths with AOSM, their mothers (n = 88, mean age 39.4 years), their fathers (n = 36, mean age 44.9 years), and community controls (n = 100 adolescents, mean age 16.5 years; n = 96 mothers, mean age 43.8 years; n = 87 fathers, mean age 45.9 years) were recruited. All participants completed a personality questionnaire and were interviewed on several measures, including structured interviews for psychopathology and substance misuse. The findings indicated that novelty seeking (NS), one of the personality dimensions, was significantly correlated with substance misuse in adolescent probands, adolescent controls, and proband fathers and mothers, but not in control fathers and mothers. Regression analyses that included conduct disorder (CD) or antisocial personality disorder (APD) symptoms indicated that both NS and CD or APD symptoms made significant contributions to the prediction of substance misuse in treatment group probands and in their fathers and mothers. The findings further suggest that NS and antisocial behaviors contribute independently to substance misuse in severely impaired adolescents and their fathers, but not in their mothers. PMID- 10365191 TI - Psychotherapies for adolescent substance abusers: 15-month follow-up of a pilot study. AB - In order to test the hypothesis that adolescent substance abusers could be matched to effective treatments on the basis of their comorbid psychopathology, 32 dually diagnosed adolescents were randomized into two short-term outpatient group psychotherapies: cognitive-behavioral treatment (CBT), and interactional treatment (IT). Two follow-up assessments were conducted at 3 and 15 months after planned treatment completion. As reported recently, at the three-month follow-up, no patient-treatment matching effects were identified. However, adolescents assigned to CBT demonstrated a significant reduction in severity of substance abuse compared to those assigned to IT. At 15-month follow-up, there were no differential improvements as a function of therapy type. However, subjects in general maintained significant treatment gains on the substance abuse, family function, and psychiatric status domains of the Teen-Addiction Severity Index (T ASI), and both CBT and IT were associated with similar long-term gains. Large scale, randomized, controlled treatment studies are further recommended to examine the findings of this small-scale pilot study. PMID- 10365192 TI - Pergolide mesylate for cocaine abuse: a controlled preliminary trial. AB - A small, controlled study was conducted to assess whether pergolide mesylate has clinical promise as a treatment for cocaine abuse prior to embarking on a larger, randomized, double-blind, controlled trial. Fourteen individuals were placed on placebo for 2 weeks, followed by a 24-week single-blind study in which they were placed on pergolide for 12 weeks, followed by placebo for 12 weeks. Another 14 patients received single-blind placebo for two weeks and then were randomized into a 24-week double-blind, placebo-controlled, multiple baseline design. Initially, patients enrolled in the study were placed on risperidone (n = 9) or placebo (n = 5). During the first 12 weeks, retention was worse for those receiving pergolide compared to risperidone or placebo. Neither risperidone nor pergolide were more efficacious in reducing cocaine use than placebo. Although earlier open studies found pergolide to show promise as a treatment for cocaine abuse, this study did not support these earlier findings. Comparing an agent to both an active control and placebo group may better predict whether a promising new agent will have clinical utility compared to the standard open trial. PMID- 10365193 TI - The disabling nature of comorbid depression among older DUI recipients. AB - Alcoholism and depression are two of the most common and disabling mental illnesses in late life. This study is a descriptive report of a sample of 49 adults who had recently been convicted of Driving Under the Influence of alcohol (DUI). A lifetime history of alcohol abuse or dependence was present in 48 subjects (98%), while a depressive disorder occurred in 24 (49%) of the subjects. Concurrent alcoholism and depression, present in 12 subjects (24.5%), produced greater self-reported disability compared to those subjects with alcoholism alone. One-year longitudinal follow-up was available on 31 subjects (63.3%). Over the course of one year, there were no changes in drinking behavior, depressive symptoms, or self-reported quality of life. These data support previous studies that suggest greater disability in patients with concurrent mental illnesses. PMID- 10365194 TI - The relationship of Axis II personality disorders to other known predictors of addiction treatment outcome. AB - This study evaluated the prevalence of Axis II disorders in substance abuse patients and the relationship between Axis II psychopathology and two other known predictors of adverse addiction treatment outcomes, i.e., Axis I psychiatric comorbidity and illegal drug use, specifically cocaine. 232 patients with cocaine and/or alcohol dependence were admitted to either inpatient or outpatient addiction recovery programs at Carrier Foundation, a nonprofit, private-pay hospital in New Jersey. Axis II disorders were more prevalent in cocaine than alcohol dependence and in patients with Axis I psychiatric comorbidity. When all three predictors were evaluated in one prediction model, the combination of Axis I and II psychopathology was the best predictor of a return to substance use at one year post-treatment, compared to the three factors alone. These findings highlighted the importance of the interrelationship of the relative prognostic value of three known predictors of addiction treatment. PMID- 10365195 TI - Heroin chasers and heroin injectors: differences observed in a community sample in London, UK. AB - "Chasing the dragon" has spread rapidly as a method of use of heroin in many countries over the last quarter of a century, and is now the most widely used method of heroin use worldwide. However, little examination has been made of the differences in characteristics and drug-taking patterns between heroin chasers and injectors. In this study, a comparison is made of the personal, social and drug-taking characteristics of heroin users in a community sample (i.e., not drawn from treatment services, contacted through a range of non-treatment access routes), according to whether the heroin user was currently a heroin chaser or injector, and according to whether or not they had ever injected. Data were examined on 400 heroin users contacted and interviewed in South London by privileged access interviewers using a structured interview schedule. Severity of dependence was measured using the Severity of Dependence Scale (SDS). Heroin chasing and injecting were almost equally prevalent among this community sample. Of the 400 heroin users, 178 (44.5%) identified "chasing the dragon" as their current route of heroin use, and 222 (55.5%) injecting. Heroin chasers were younger, though they had first used at an older age, had a larger proportion of non-using friends, and contained larger proportions of women and people of Afro Caribbean origin. Injectors were using higher daily doses and were significantly more likely to be using on a daily basis. Those who had never injected were more likely to be women and to have friends who did not use drugs. Severity of Dependence (SDS) scores were greater amongst injectors, with scores for injectors almost all being above the informal clinical threshold of greater than 5; while one third of the chasers had SDS scores below this level. Heroin chasing had been a longstanding pattern of behavior for a large proportion of the study sample. Chasers were generally less deeply involved in a heroin-using culture and were less likely to be using heroin daily. Almost all low scores for severity of dependence were seen among the chasers, though a significant number of chasers had dependence scores at the most extreme. Heroin chasers display distinctly different personal and drug-taking characteristics and will need to be the subject of separate research studies and clinical programs. PMID- 10365196 TI - Nalbuphine hydrochloride dependence in anabolic steroid users. AB - Nalbuphine hydrochloride, a nonscheduled opioid agonist/antagonist analgesic, is currently approved for the treatment of pain. Recently, nalbuphine dependence was reported in three anabolic steroid users in Britain. To further document this phenomenon, we conducted interviews on eleven subjects who reported nalbuphine use. Eight subjects were clinically dependent on nalbuphine, and seven of the subjects who were asked about tolerance and withdrawal with nalbuphine acknowledged these symptoms. Eight subjects, who had never used drugs intravenously before, reported using nalbuphine by this route. Nalbuphine-related morbidity was extensive and included medical complications and psychiatric symptoms. Nalbuphine users also exhibited a high rate of comorbid Axis I disorders, including other substance misuse. Virtually all subjects described widespread nalbuphine use in the gymnasiums they frequented. These observations, together with the recent increase in nalbuphine-related articles in the lay press, suggest that nalbuphine may represent a new drug of abuse among athletes, especially those using anabolic steroids, and that nalbuphine's scheduling status may need to be re-evaluated. PMID- 10365197 TI - Cocaine-induced mood disorder: prevalence rates and psychiatric symptoms in an outpatient cocaine-dependent sample. AB - This paper attempts to examine and compare prevalence rates and symptom patterns of DSM substance-induced and other mood disorders. 243 cocaine-dependent outpatients with cocaine-induced mood disorder (CIMD), other mood disorders, or no mood disorder were compared on measures of psychiatric symptoms. The prevalence rate for CIMD was 12% at baseline. Introduction of the DSM-IV diagnosis of CIMD did not substantially affect rates of the other depressive disorders. Patients with CIMD had symptom severity levels between those of patients with and without a mood disorder. These findings suggest some validity for the new DSM-IV diagnosis of CIMD, but also suggest that it requires further specification and replication. PMID- 10365198 TI - Methadone overdose in an opiate-naive patient. PMID- 10365199 TI - [Intratracheal intubation without muscle relaxant with the use of remifentanil propofol]. AB - OBJECTIVES: To assess tracheal intubation conditions after induction of anaesthesia with remifentanil and propofol, using itemized scoring criteria. STUDY DESIGN: Clinical, prospective, open, non comparative trial. PATIENTS: One hundred consecutive patients undergoing surgery not requiring muscle relaxation, during the study period extended over 12 months. METHODS: After premedication with lorazepam (2 mg) the day before and hydroxyzine (100 mg) one hour before surgery, anaesthesia was induced with remifentanil administered continuously with a syringe pump at a rate of 1.20 +/- 0.06 micrograms.kg-1.min-1 and propofol (3 mg.kg-1 IV bolus). The trachea was intubated two minutes later and mouth opening, glottis exposure, glottis opening, movements, additional anaesthetic agents and chest rigidity were recorded. RESULTS: Intubation conditions were excellent in 87% of patients, and the tube was inserted rapidly, within two minutes. However in 38% of patients the cuff inflation caused cough. In 13%, glottis opening was delayed and intubation required three minutes. A major decrease of arterial pressure and heart rate was recorded in 9 and 6% of patients respectively. CONCLUSION: Induction of anaesthesia using remifentanil and propofol allows satisfactory tracheal intubation without a muscle relaxant. However this technique is contraindicated: a) in patients with a full stomach, as intubation is not always successful at the first attempt; b) in patients scheduled to undergo neurosurgery or ophthalmic surgery, as tracheal intubation may elicit cough, increasing intra-cranial and intra-ocular pressure; c) in patients in poor circulatory status, as it decreases significantly arterial pressure and heart rate. PMID- 10365200 TI - [Severe head injuries: effects of pre-hospital mechanical ventilation on capnia]. AB - OBJECTIVE: To assess the effect on PaCO2 of mechanical ventilation during prehospital management of severely head-injured patients. STUDY DESIGN: Retrospective observational study. PATIENTS: Severely head-injured patients with Glasgow coma score < or = 8. All patients were sedated, with the trachea intubated and the lungs mechanically ventilated. METHODS: According to the capnia measured at the admission in the neurosurgical intensive therapy unit they were allocated into one of the following three groups: hypocapnia group (PaCO2 < 30 mmHg), recommended capnia group (PaCO2 = 30-38 mmHg) and hypercapnia group (PaCO2 > 38 mmHg). RESULTS: Out of the 42 patients with similarly severe head injuries, 19% were included in the recommended capnia group (PaCO2: 34 +/- 2 mmHg), 38% in the hypocapnia group (PaCO2: 23 +/- 3 mmHg) and 43% in the hypercapnia group (PaCO2: 47 +/- 7 mmHg). In all except three, PaO2 was above 95 mmHg. The settings of ventilatory parameters on the ventilators were similar. CONCLUSION: In 81% of patients, mechanical ventilation was inadequate as far as PaCO2 levels are concerned. Major hypocapnia and hypercapnia carry a potential risk for cerebral ischaemic. Therefore it is recommended to monitor PETCO2 during prehospital transport in medical ambulances and to determine arterial blood gases at arrival of severely head-injured patients in the admission unit for emergencies. PMID- 10365201 TI - [Evaluation of an activity score of prehospital medicine: activity scoring using Smur (CAS)]. AB - OBJECTIVE: The activity of prehospital emergency medicine (PEM) teams is mainly assessed by the amount of medical interventions and their duration. However, these two parameters do not reflect workload correctly. The aim of this study was to compare a new activity scoring system for PEM teams (CAS) with the standard TISS score. STUDY DESIGN: Prospective comparative study. PATIENTS: This study included 4,650 patients, with a median age of 39 years [0-100], attended by PEM teams during 4,189 ambulance transports (83% primary transports and 17% interhospital transfers). METHODS: The CAS score derived from Omega scoring system is the sum of 51 items specifically suited for PEM, each one being rated 1, 3, 6 or 10. CAS and TISS, were prospectively determined for all medical ambulance transports over 1.5 year. Transport data and main diagnosis were collected. Results were analysed with non parametric statistical tests. RESULTS: Median duration of interventions (39 min [0-475]) and median CAS score (7 [0-72]) were comparable (R' = 0.38, P < 0.001). Median TISS was 3 [0-30]. CAS score was correlated with TISS (R' = 0.92, P < 0.001). CAS score was higher in men than in women (7 [0-72] vs 7 [0-45], P < 0.001). CAS scores in patients with cardiologic (11 [0-72]), respiratory (9 [0-31]) or neurological insults (9 [0-43]) were significantly higher than those of patients with other insults (P < 0.001). CONCLUSION: The CAS scoring system is a valuable indicator of medical team prehospital workload, probably more suited for prehospital emergency medicine than TISS. PMID- 10365202 TI - [Patient-controlled sedation with propofol for extracorporeal shock wave lithotripsy]. AB - OBJECTIVE: Evaluation of patient-controlled sedation with propofol for extracorporeal shock wave lithotripsy (ESWL) using an EDAP LT01 lithotriptor. STUDY DESIGN: Prospective clinical study. PATIENTS: Fifty consecutive patients, ASA I or II, aged 18-65 years. METHODS: Patients received 50 mg of propofol five minutes before ESWL, then they self-administered 50 mg bolus doses of propofol with a ten minutes lock-out interval. Pain (1-100 mm VAS) and sedation (four points scale) were assessed every five minutes. Patient satisfaction was recorded at the end of the procedure. Pharmacokinetic simulation was done with the Marsh's data set. RESULTS: Three patients were excluded. Patients received a mean propofol dose of 147 +/- 68 mg during the procedure with a mean duration of 47 +/ 8 minutes. The median of the higher sedation scores was 2 (drowsy) and mean maximal VAS was 40 +/- 20 mm (10-70). No complications were recorded. Thirty-nine patients (83%) were satisfied or very satisfied by patient-controlled sedation. CONCLUSION: Patient-controlled sedation with propofol is a safe and efficient mode of administration of an hypnotic agent for ESWL. PMID- 10365203 TI - [Effects of anesthetic agents on arterial reactivity]. AB - OBJECTIVE: To review the effects of halogenated and intravenous anaesthetics on arterial vasoreactivity. DATA SOURCE: Articles were obtained from a MEDLINE review (search terms: 'vascular smooth muscle, endothelium' used separately or associated with following anaesthetic agents: 'halothane, isoflurane, enflurane, desflurane, sevoflurane, thiopentone, propofol, ketamine, etomidate'. Other sources included review articles and textbooks. STUDY SELECTION AND DATA EXTRACTION: All experimental studies published since 1975 were analysed and pertinent data extracted. DATA SYNTHESIS: Within the vascular wall, arterial vasoreactivity involves the endothelium and the vascular smooth muscle. In vivo, arterial vasoreactivity is regulated by neuronal, hormonal, and metabolic factors. In vitro, the direct action of anaesthetic agents on the vessel can be studied in the absence of such factors. In vitro studies with arterial rings have shown that inhalational anaesthetics directly decrease endothelium-independent contraction induced by various pharmacological agents. This direct effect of anaesthetics results from a decrease in intracellular calcium, mainly caused by an inhibition of transsarcoplasmic calcium influx. Volatile anaesthetics decrease endothelium-dependent vasorelaxation at a site(s) within the nitric oxide (NO) signalling pathway, located downstream from the NO-related receptors and upstream from guanylyl cyclase. They may also decrease endothelium-independent vasorelaxation by inhibiting NO activation of guanylate cyclase. Intravenous anaesthetics, such as propofol, barbiturates, ketamine and etomidate also decrease vasoconstriction by various degrees. Propofol is the most potent inhibitor of vasoconstriction and thiopental the least one. All these IV anaesthetics have been shown to inhibit in some circumstances both endothelium dependent and -independent vasorelaxation. Further studies are required to enable a better understanding of the mechanism and the site of action of these vascular effects of anaesthetics. For example, the investigation of the effects of anaesthetic agents on vascular reactivity in diseases associated with endothelial dysfunction may indirectly provide insight into the role of endothelium. PMID- 10365204 TI - [Sheehan's syndrome: a recurrent obstetrical complication]. AB - Case report of a 25-year-old primigravida who sustained a necrosis of the anterior pituitary gland after a haemorrhagic Caesarean section. The diagnosis was delayed, as the early symptoms (apathy, anorexia, nausea, orthostatic hypotension and hypoglycaemia) were imputed to more usual causes. PMID- 10365205 TI - [Accidental intracerebral penetration of a nasal hemostatic probe]. AB - We report a case of inadvertent intracerebral introduction of a haemostatic device (Brighton tube) inserted into a nasal cavity for control of epistaxis in a patient with major craniofacial trauma. This complication remained unrecognized in the unconscious patient until the subsequent CT-scan control. In unconscious patients with a major facial trauma, intranasal haemostatic probes should be inserted under direct visual control by a ENT specialist and their position checked by digital palpation of the inflated cuffs behind the soft palate. PMID- 10365206 TI - [Right subclavian catheterization. Misplaced insertion into the subclavian artery and the ascending aorta]. AB - The accidental subclavian artery puncture is usually obvious. We report a case of unrecognized arterial catheterisation. The catheter had been inserted during anaesthesia after return of dark and non pulsatile blood, and not controlled by a chest radiograph. During surgery, the injection of 40 mL isotonic saline containing 4 g of piperacillin for antibiotic prophylaxis resulted in a transient circulatory collapse associated with ECG tracing of myocardial ischaemia. Postoperative chest radiograph showed that the catheter was in a midsternal position, at the level of the ascending aorta. The intracoronary penetration of piperacillin was considered as the cause for the transient cardiocirculatory changes. The various diagnostic tools of the intra-arterial location of the catheter are discussed. All inadvertent subclavian artery catheterisations published in the literature have been carried out with multi-lumen catheters. The latter can contribute to the failure to recognize the arterial puncture and catheter insertion because of the use of a small bore needle (Seldinger's technique) and infusion with electrical pumps. PMID- 10365207 TI - [Severe lactic acidosis and thiamine deficiency during parenteral nutrition in a child]. AB - We report the case of a leukemic child treated with chemotherapy and parenteral nutrition for three weeks, who developed a severe lactic acidosis. Clinical features included both digestive and neurological disorders associated with a moderate cardiovascular collapse. After elimination of a toxic, a neoplastic or a septic cause, a thiamin (or vitamin B1) deficiency was suspected because of the lack of vitamin supply to parenteral nutrition. Intravenous administration of thiamin rapidly controlled lactic and clinical features. The diagnosis was confirmed by a low plasmatic concentration of thiamin. Thiamin deficiency must be suspected in case of severe lactic acidosis during parenteral nutrition and systematically prevented by supply of vitamins. PMID- 10365208 TI - [Treatment of severe cardiogenic pulmonary edema with continuous positive pressure ventilation using a cuffed cannula Copa]. AB - We report the use of a cuffed oropharyngeal airway (Copa), in a patient with an acute respiratory failure from a cardiogenic pulmonary oedema, for continuous positive pressure ventilation. Considering the ease of use and the lack of laryngeal stimulation, this device can be considered for mechanical ventilation in selected cases with acute cardiac failure. There are two contra-indications: prolonged mechanical ventilation, because of the lack of airway protection from gastro-oesophageal reflux, and normal consciousness, as the patient cannot swallow. This device can be considered when starting intensive therapy including mechanical ventilation in patients with acute respiratory failure of foreseen short duration. PMID- 10365209 TI - [Barium sulfate poisoning?]. AB - We report a case with neurologic symptoms which occurred after iterative radiologic examinations of the gastrointestinal tract with barium sulphate, which were related to a barium encephalopathy. Suspected by the presence of barium in the blood, a systematic intoxication was occurred without any evidence for a gut or a vascular leak. This case raises the question about a possible extraluminal diffusion of such a heavy metal, given orally, in normal conditions of use. PMID- 10365210 TI - [Pheochromocytoma. Severe and uncommon presentations]. AB - We report five cases of phaeochromocytoma in patients admitted for myocardial infarction, severe cardiac failure, with shock, stroke and ischaemic gangrene of a lower limb respectively. The pathophysiology of these events is discussed. Early surgery prevents visceral damage from massive release of catecholamines. PMID- 10365211 TI - [Anesthetic apparatus: failure of fresh gas delivery to the auxiliary circuit]. AB - A case of sudden loss of fresh gas delivery to a manual auxiliary anaesthetic circuit is reported. The corresponding fresh gas outlet was disunited from the anaesthesia machine and had been inadvertently rotated in its housing, allowing the rubber tubing linking the fresh gas delivery unit to the gas outlet to be twisted. PMID- 10365212 TI - [Intraoperative problems with cardiac rhythm in relation to hypokalemia and the administration of atropine]. PMID- 10365213 TI - [Spinal anesthesia and von Bezold-Jarisch reflex?]. PMID- 10365214 TI - [Microgram should read microgram and not mcg!]. PMID- 10365215 TI - [Gamma-OH: a comeback]. PMID- 10365216 TI - [How I effected a brachial plexus block for shoulder arthroscopy]. PMID- 10365217 TI - [Patient information and follow-up of transfusions: "a checked report"?]. PMID- 10365218 TI - [Practical information about material vigilance in anesthesia and recovery. 3]. PMID- 10365219 TI - [Post-traumatic pneumomediastinum]. PMID- 10365220 TI - [The bicentennial of the death of Luigi Galvani, initiator of the study of electrophysiology]. PMID- 10365221 TI - Captopril and glutathione inhibit the superoxide dismutase activity of Hg (II). AB - The purpose of this study was to determine if captopril, an angiotensin converting enzyme inhibitor, and glutathione could interact with mercuric ions and so modify the catalytic dismutation of superoxide carried out by this metal. With an assay that generates superoxide anion radicals without the intervention of metal ions increasing concentrations of both reagents progressively inhibited the breakdown of superoxide brought about by mercury. Maximum inhibition was attained with a molar ratio of captopril (glutathione): Hg (II) = 1. These results may help to explain the protective and/or antioxidative effect of thiol compounds during mercury intoxication. PMID- 10365222 TI - [Late thrombolysis and electrical stability in acute myocardial infarction. Role of the collateral flow]. AB - We evaluated 249 patients (pts) with first acute myocardial infarction: 1. Pts without thrombolysis, n = 119, 2. Pts treated with thrombolysis within 6 hours following MI, n = 80 and 3. Pts treated with thrombolysis between 6-12 hours after MI. Arrhythmic events were evaluated during follow up. All underwent heart rate variability studies and coronary angiogram where anterograde flow (TIMI) and collateral flow (Rentrop scale 0-2 = poor collateral flow and 3 = good collateral flow) were determined. Pts in group 2 and 3 showed a better anterograde and collateral flow than group 1 (p < 0.001). A lower spectral power in the high frequency band and a higher ratio low/high frequency band were observed in group 1 (p < 0.05). Conjunctive consolidation analysis showed more malignant arrhythmias in TIMI 0-2 with poor collateral flow than TIMI 0-2 with good collateral flow (17/138-12.3% vs 0/14-0%). Kaplan Meier analysis was able to demonstrate more cardiac sudden death events in TIMI 0-2 with poor collateral flow than TIMI 0-2 with good collateral flow or TIMI 3 (x2 = 7.22, p = 0.028), independently of thrombolytic treatment. PMID- 10365223 TI - [The utility of rapid qualitative determination of troponin T, the MB fraction of creatine phosphokinase and myoglobin in acute ischemic coronary syndromes]. AB - The objective of our study was to validate the diagnostic utility of cardiac troponine T in acute ischemic syndromes, and also in cases of difficult diagnosis. We analyzed its concordance and compare them with conventional enzymatic quantitative methods. We determined sensitivity, specificity, positive and negative predictive values and likelihood ratio. Kappa index was used to know the concordance grade between T troponin and the positive or negative results of the quantitative enzymatic curve. Stochastic significance was valued by Chi square of Mcnemar test. In seventy patients who arrived to the hospital with chest pain who were assigned to five different groups. The sensitivity in quantitative markers was higher than qualitative methods, however the specificity, likelihood ratio was lower. In the total group the concordance analysis between qualitative and quantitative markers was adequate, (kappa index 0.65 p < 0.05). This study suggest that the rapid bedside qualitative test by cardiac Troponin T is a good diagnostic marker compared with conventional quantitative markers to evaluate chest pain in acute ischemic syndromes. PMID- 10365224 TI - [The effectiveness and safety of d,l-sotalol in the ambulatory treatment of atrial fibrillation and flutter]. AB - Data on short and long term efficacy and safety of d,l sotalol in patients with atrial fibrillation or atrial flutter is limited. The aims of this study were to (1) assess the antiarrhythmic efficacy of d,l sotalol maintaining normal sinus rhythm in patients with refractory atrial fibrillation or flutter, (2) evaluate the efficacy of d,l sotalol in preventing recurrences of paroxysmal atrial fibrillation or flutter, (3) evaluate the control of ventricular rate in patients with paroxysmal or refractory atrial fibrillation or flutter unsuccessfully treated with other antiarrhythmic agents, (4) determine predictors of efficacy (5) assess the safety of d,l sotalol in this setting. Two hundred patients with chronic or paroxysmal atrial fibrillation or atrial flutter or both, who had failed one to six previous antiarrhythmic drug trials were treated with d,l sotalol 80 to 440 mg/day orally. Fifty four percent was female, age 47 +/- 16 years (range 7-79), follow up period 7 +/- 7 months (range 1 to 14 months), 79% of patients had the arrhythmia for more than one year. The atrial fibrillation in 37.5% of patients was chronic and paroxysmal in 23.5. The atrial flutter was chronic in 31% of patients and paroxysmal in 8%. Eighty two percent of patients was in functional class I (NYHA) and 82% had cardiac heart disease: left atrial (LA) size 44 +/- 10 mm, right atrial (RA) size 37 +/- 7 mm and left ventricular ejection fraction (LVEF) 58 +/- 8%. Total success was achieved in 58% of patients (atrial fibrillation 40% and 18% in atrial flutter), partial success in 38% (atrial fibrillation in 18% and 20% in atrial flutter) and 4% of patients failure. It was p < 0.07 when compared total success vs partial success among atrial fibrillation and atrial flutter groups. Patients with cardiac heart disease responded worst (p = 0.10) to the drug than those without it, specially if the heart was dilated. We concluded that d,l sotalol has moderate efficacy to convert and maintain normal sinus rhythm, as well as it acts controlling paroxysmal relapses and ventricular heart rate. PMID- 10365225 TI - [A comparison of clinical and angiographic outcome after implantation of a Wiktor stent in large and small coronary arteries]. AB - The objective of this study was to investigate the mid term influence of vessel size in clinical and angiographic outcome, after Wiktor stent implantation in arteries larger and smaller than 3 mm. METHOD: A total of 188 stents were implanted in 167 patients divided in two groups. Group 1: stents implanted in arteries smaller than 3 mm, 40 stents in 38 patients. Group 2: in arteries larger than 3 mm. 148 stents in 129 patients. Clinical follow up and a repeated coronary angiographic study were carried out after six months. RESULTS: Angiographic success was achieved in 97% and 98% cases, with clinical success in 92% and 95% respectively. Acute occlusion occurred in 2 patients of group 1 (5%), and in four patients in group 2 (2.7%); one patient died and four patients suffered a non fatal myocardial infarction. During clinical follow up, nine patients presented a major complication, two in group 1 and seven in group 2 (5.5% vs. 5.6%). Asymptomatic survival was 86% and 84% respectively. In angiographic follow up we observed a restenosis rate of 41% of the patients in group 1 and 25% of those in group 2. Immediate gain was similar in both groups, but late loss (1.06 +/- 0.85 vs 0.97 +/- 0.86) and loss rate (0.60 vs 0.46) were greater in group 1. CONCLUSION: The frequency of stent thrombosis as well as the incidence of restenosis were higher in arteries smaller than 3 mm. No differences were observed in the incidence of major ischemic events. PMID- 10365226 TI - [Torsades de Pointes: a cause of syncope in sinus node dysfunction]. AB - Torsade de Pointes (TdP) is an atypical ventricular tachycardia that occurs in the setting of QT interval prolongation which may be an inherited or acquired abnormality. TdP is a recognized complication of bradyarrhythmias, specifically atrioventricular block. However, sinus node dysfunction is a rare cause of syncope associated with TdP. Experimental studies have suggested that the acquired long QTU interval and TdP may be due to bradycardia or pause-dependent early after depolarizations that give rise to triggered activity. This report describes a patient with sinus bradycardia and intermittent AV nodal rhythm induced long QT interval and repetitive syncope related to TdP and successfully treated with temporal and long-term pacing. PMID- 10365227 TI - [Myocardial bridging. The role of microcirculation, coronary reserve and systolic endothelial injury]. AB - BACKGROUND: The relationship between myocardial bridging (MB) and ischemic heart disease is still controversial. However, a recent new evidence suggests that this relation is not by chance. PURPOSE: The purpose of our study was to review in a critical manner, the evidence for the relationship between MB and myocardial ischemia and its possible consequences. METHODS: We present 2 cases of our series and review the medical literature from January 1966 to January 1998 published and included in Medline and Current Contents. RESULTS AND CONCLUSIONS: The principal findings after this review were: 1) MB is not a normal variant; 2) The clinical impact of MB depends on its anatomical extension and degree of compressive effect; 3) The MB muscle is not similar to myocytes from other cardiac areas; 4) The environment surrounding coronary artery may be a crucial factor in determining whether the MB influences the induction of heart disorders or not; 5) The overshoot due to compressive effect on coronary artery might determine endothelial injury in the microcirculation post-MB; 6) In some cases, the systolic endothelial injury may contribute to release factors that are able to reduce the coronary reserve, resulting in myocardial ischemia; 7) The possible role of PTCA in this disorder still has to be proven. Surgical treatment should be considered when important myocardial ischemia had been demonstrated, even in those asymptomatic cases. PMID- 10365228 TI - [Tachycardiomyopathy in patients with and without subacute cardiac pathology. Experiences at the Cardiovascular Center of Merida, Venezuela]. AB - We report our experiences with tachycardia-induced cardiomyopathy. Nine patients (3-56 years old) had incessant supraventricular tachycardia and congestive heart failure. The cardiac eco-Doppler evidenced a significant increase of cardiac volumes and mild tricuspid and mitral regurgitation. The ejection fraction (EF) was 0.31 +/- 0.12, the end diastolic volume was 162 +/- 48 cc and the end systolic volume, 116 +/- 54 cc. Four patients had accessory pathways, 3 atrial flutter, 1 A-V nodal reentrant tachycardia, and 1 ectopic atrial tachycardia. Two patients had Chagasic myocarditis. Only in one chagasic patient a decreased number of tachycardia episodes was achieved, this patient died. The autopsy revealed cerebellar and pulmonary emboli. In the other 8 patients the arrhythmia was well controlled. In these, the ventricular volumes decreased, the EF increased to 0.51 +/- 0.14 (p = 0.00006), and the congestive heart failure remitted. We conclude that incessant tachycardia produces a symptomatic dilated cardiomyopathy in patients with and without structural heart disease. The arrhythmia control is followed by an increase in cardiac function and a remission of heart failure symptoms. PMID- 10365229 TI - [Critical pulmonary valve stenosis in neonates; treatment with balloon valvuloplasty using an arteriovenous loop and gradual dilatations]. AB - Critical pulmonary stenosis of the newborn is an emergency and the balloon dilatation can represent its definitive treatment. This procedure has important technical aspects related with the age and weight of patients and to anatomical factors that contributes to the obstruction. We present the case of a 20 days old newborn with this problem, in whom the use of an arterio-venous loop through the ductus arteriosus allowed the passage of balloon catheters of gradually increasing diameter until a balloon/pulmonary annulus index of 1.4 was reached. This procedure can be performed safely in cases in whom the common technique is difficult to apply. The procedure allows a successful valvular dilatation. PMID- 10365230 TI - [Myocardial reperfusion injury. Its impact on clinical practice. II]. PMID- 10365231 TI - [Supraventricular tachycardia. Drugs or ablation?]. PMID- 10365232 TI - [Design and development of a stent (SAQ) for the treatment of arterial coronary arterioscleotic obstruction]. AB - The practice of percutaneous transluminal coronary angioplasty has shown that the major complications are acute dissection as well as suboptimal results and restenosis. The effort to reduce these complications has led to create an intravascular device called Stent. The technology is complex and very expensive, for this reason we designed and made a new model of stent named SAQ. We introduce: The methodology of development, fabrication and modifications of a new intravascular device Stent SAQ. The results obtained in coronary arteries of ex vivo hearts of pigs and humans. The results in two model of animals, rabbit aorta and peripheral arteries in dogs. This investigation at this phase, shows satisfactory properties of SAQ which is secure and effective, with similar properties to the stents in use. PMID- 10365233 TI - [Fenestrated Fontan surgery in high risk patients]. AB - A retrospective analysis is presented of all patients who had fenestrated Fontan procedure between january 1990 and may 1996. Surgery was indicated in the presence of anyone of the following risk factors: mean pulmonary pressure higher than 20 mmHg; pulmonary vascular resistance higher than 2 UW; ejection fraction less than 60%; systemic ventricular end diastolic pressure higher than 8 mmHg; Nakata index less than 200 mm2/m2, McGoon index less than 2. The diagnosis were: Absent right atrio-ventricular connection with concordance ventriculo-arterial connection 10 patients; pulmonary atresia with intact septum, 1 patient; Ebstein's malformation, 1 patient and absent left A-V connection with discordance VA connection, 1 patient. The mean of age was 6.7 years (range 2.5-11 years). Overall mortality was 23%. No significant difference in risk factors was found between survivals and no survivals. Nonsurvivors had between two an four risks factors. Postoperative complications were 1 patient with protein losing enteropathy and stroke (1 patient). The mean duration of pleural effusion was 16 days (range 4-45 days). We consider fenestrated Fontan procedure useful for patients with congenital heart disease with a one hypoplastic ventricle and one o more risks factors. PMID- 10365234 TI - [Coronary revascularization without extracorporeal circulation. An alternative therapy (preliminary report)]. AB - Direct myocardial revascularization techniques has earned great acceptance in the treatment of ischemic coronary syndromes in the past two decades. Almost since its beginnings the procedure was done with the aid of extracorporeal circulation, since the technical accessibility with cardiac standstill further helped to evolve the procedure. As years went by, complications due to the use of the pump became evident, such as microembolism, bleeding disorders, and others. Besides this, there is a special group of patients, those with high surgical risk, with preexisting pulmonary, hepatic or cerebral disease, who benefit a great deal when bypass grafting procedure is done without the pump. In this article we delineate the indications, contraindications, and the procedure itself based in our personal experience, reporting our first five patients with this technique. PMID- 10365235 TI - [Automatic, implantible cardioverter-defibrillator in a patient with chronic Chagas cardiopathy and sustained ventricular tachycardia]. AB - We studied a 48 years old woman, with chronic Chagasic cardiopathy, manifested with cardiomegaly, heart failure and syncope, due to a sustained ventricular tachycardia (SVT) of two different configurations (left bundle branch block and right bundle branch block). During electrophysiological testing, both types of ventricular tachycardia were reproduced. Successful ablation therapy of the right branch of His was performed due to suspicion of the bundle branch reentrant tachycardia, with a left bundle branch block. The patient continued to show SVT episodes, now with right bundle branch block pattern. Cardioverter Defibrillator was implanted. We report this case due to the rare frequency of Chagas' disease, where it could be a cause of heart disease, since the existence of the parasite (trypanosoma cruzi) and its vector (Triatoma) has been identified in some rural and suburban zones in the state of Aguascalientes, Mexico. PMID- 10365236 TI - [Primary angioplasty: comparison of working hours and night]. AB - A prospective, observational, comparative study of 100 patients with acute myocardial infarction and primary angioplasty was performed to establish if there was statistically difference between the lag of time when symptoms begin and the time of the emergency admission to the time of arrival at the catheterization suite during working hours vs the "on call" hours. Patients were allocated in two groups accordingly to morning hours or on call hours. Time of onset of symptoms to the catheterization suite arrival between the two groups was no significantly different. Time from emergency room arrival to catheterization suite arrival was significant different < 0.05, however success rate between groups 86% vs 80% and complications rate were statistically non significant between both groups. We conclude that primary angioplasty is a highly effective method of reperfusion. Even though the time from the emergency room arrival to the catheterization suite arrival was significantly less during day than the on call hours, there is no difference between the success rate and complications incidence in both groups. PMID- 10365237 TI - [Clinical, parasitological and histopathologic follow-up studies of acute Chagas patients treated with benznidazole]. AB - With the purpose of studying their clinical and histopathologic evolution, 10 acute chagasic patients with myocarditis diagnosed by endomyocardial biopsy and positive sero-parasitologic methods were evaluated at 11 months (8-21 months) after treatment with oral benznidazole. Four of them were reevaluated 5 years post-treatment (58-68 months). Study protocol consisted of clinical, hemodynamic, echocardiographic, seroparasitologic and histopathologic evaluations. Results showed evidence of persisting myocarditis in 90% and 75% of patients evaluated at 11 months and 5 years respectively, along with asymptomatic, subclinical left ventricular systolic dysfunction being recognized in 75% of patients evaluated 5 years after treatment. All parasitologic studies became negative during follow up, but serology remained positive for Trypanosoma cruzi antibodies in 80% and 75% of patients studied at 11 months and 5 years. In conclusion, myocardial damage was constantly found in our acute chagasic patients. Treatment with benznidazole eliminated symptoms and parasitemia, but it does not seem to alter favorably the histopathological evolution of the chagasic cardiac disease. PMID- 10365239 TI - [Neonatal cardiac rhabdomyoma: case report and clinico-epidemiologic considerations]. AB - Primary cardiac tumors are very infrequent at all ages; the most frequent in the pediatric age is rhabdomyoma. This tumor is associated with tuberous sclerosis in 37 to 80%, with a frequency of 1 for each 40,000 live newborns. This case is about a newborn, who in the immediate postnatal period presented pansystolic murmur, grade IV cardiomegaly, electrocardiographic changes of biventricular hypertrophy and heart failure. Echocardiogram and magnetic resonance images showed several tumors in the septum and in ventricular walls; histopathology study of the heart, confirmed the diagnosis. The diagnosis of tuberous sclerosis was made clinically (seizures, hypomelanotic macules) and with the image of parenchymal hypodense areas. In our country there is little information about both diseases, that's why we made a review of the incidence, diagnosis, prognosis and treatment. PMID- 10365238 TI - [Results of treatment of acute myocardial infarction with thrombolytic therapy. Experiences in 437 patients in the coronary care unit of the National Institute of Cardiology "Ignacio Chavez"]. AB - OBJECTIVE: To review the results and complications of thrombolysis in patients with acute myocardial infarction (AMI) and its complications. METHODS: Since june 1989 to august 1994 we studied patients with AMI, who underwent thrombolysis. Clinical characteristics, complications and angiographic results are described. RESULTS: Of the total population 86.3% patients received Streptokinase (SK) and 13.7% recombinant tissue plasminogen activator (rt-PA). In 20 patients the age was under 40 years, 373 between 40-70 years, and 80 patients over 70 years. 84% were men and 16% women. 72% had smoking habit; 21% diabetes mellitus, 43% hypertension, 54% had previous angina and previous AMI in 22%. The location of AMI was anterior in 234 patients and 239 inferior. In 63% enzyme washout was observed, and rapid electrocardiographic evolution in 81%. Postthrombolisis arrhythmias was observed in 64.7%. Major bleeding in 11.8% and central nervous system hemorrhage in 0.4% only with rt-PA. Postinfarction angina in 22%, and re infarction in 4%. Cardiac rupture in 1.4%, with shock and death. Mitral insufficiency in 2.1% demonstrated by echocardiogram. Coronary angiography was done in 373 patients (80%), of which 50.7% was made in the first 5 days. The culprit artery was anterior descending in 273 patients and right coronary in 95. Left ventricular dysfunction was seen in 23% in patients with anterior AMI, and 5% with inferior AMI. Cardiogenic shock was seen in 7%. Coronary artery bypass grafting was undertaken in 106 patients and coronary angioplasty in 67. The ten days mortality was 8.8%, principally due to cardiogenic shock, ventricular arrhythmias and ventricular rupture. CONCLUSIONS: The usefull permeability in the culprit artery was obtained in 40%, who had coronary angiography done 145 hours posthrombolysis. Mortality was under 10% in this study. PMID- 10365240 TI - [Use of nitric oxide in the evaluation of pulmonary arterial hypertension]. AB - The purpose of this article is to describe a patient with severe pulmonary artery hypertension, who was evaluated in the catheterization laboratory with the use of nitric oxide to check the degree of reversibility of the pulmonary hypertension. The patient is a 18 months old baby with atrio-ventricular canal and severe pulmonary hypertension, whose vascular resistance dropped from 8.75 Wood U/m2 to 1.32 Wood U/m2. With these findings the pulmonary artery hypertension was considered reversible and made him a good candidate for successful corrective surgery. The use of nitric oxide is very useful for the evaluation of the degree of reversibility of the pulmonary vascular resistance in cases with severe pulmonary artery hypertension with a left to right shunt. PMID- 10365241 TI - [Myocardial reperfusion injury: its clinical impact. I]. PMID- 10365242 TI - C-terminal region of hTPO is important for secretion and expression in insect cells. AB - Human thrombopoietin (hTPO) variant cDNAs truncated in the C-terminal regions of wild-type hTPO (332 amino acids) were constructed by PCR and expressed in Trichoplusia ni (Tn5) insect cells using a baculovirus expression system. Each variant, hTPO163 (amino acids 1-163), hTPO198 (1-198) and hTPO245 (1-245), was produced in insect cells with very low efficiency in comparison with wild-type hTPO. Immunoblot analysis showed that the predicted 20, 25 and 34 kDa molecular sizes corresponding to hTPO163, hTPO198 and hTPO245, respectively, were barely detected in culture medium and most of the proteins remained within the cell. These results suggest that C-terminal regions containing potential N glycosylation sites of hTPO are required for the secretion of hTPO into culture medium as well as expression in insect cells. PMID- 10365243 TI - Phenotypic selection: a successful strategy to fix major genes of hypertension. AB - Hypertension is dominantly inherited in cross hybrids between hypertensive SHR/Mol and normotensive BB/OK rats. We used these cross hybrids for repeated backcrossing of selected hypertensive animals onto normotensive BB/OK rats to fix high blood pressure and to generate a hypertensive and diabetic BB/OK rat subline. After 8 backcrosses, the backcross parents were genetically analysed with the aid of 259 microsatellite markers to identify SHR genes causing blood pressure of 177 +/- 10 mmHg in this BB/OK rat subline. Loci on chromosomes 1, 14 and 18 showed longest heterozygosity. These loci might contain major genes of the SHR rat causing hypertension in this BB/OK rat subline. This classical strategy seems to be most suitable to fix major genes of hypertension in particular and complex traits in general and therefore to generate new animal models. PMID- 10365244 TI - Influence of atropine on carbachol dual effect on Ca2+ mobilization in SH-SY5Y neuroblastoma cells. AB - The muscarinic receptor stimulated mobilisation of calcium ions in SH-SY5Y neuroblastoma cells was measured as a function on carbachol and atropine concentrations. The combined application of this pair of muscarinic agonist and antagonist yielded a set of bell-shaped dose-response curves. In the presence of atropine the cell responses were smaller and the up-going phase of these relationships was shifted towards higher agonist concentration, while the down going phase of these curves was not influenced by the antagonist. These results pointed to a similar mechanism of the receptor inhibition at high carbachol (agonist) concentrations and by atropine (antagonist). PMID- 10365245 TI - Kinetic properties of ATP-dependent phosphofructokinase from grapefruit juice sacs: effect of TCA cycle intermediates. AB - Grapefruit juice sac ATP-PFK was studied kinetically for its substrates ATP and Fru-6-P at pH = 7.5. The Km for ATP is equal to 39.8 +/- 4.6 microM. ATP becomes inhibitory at concentrations above 80 microM. The Km for ATP is not affected by the addition of citrate (10 mM). For Fru-6-P, the saturation curve is sigmoidal, with an S0.5 equal to 0.17 +/- 0.03 mM, in the presence of Mg++ (2.5 mM) and ATP (1 mM). ATP-PFK shows a negative cooperativity at lower concentrations of Fru-6-P (h = 0.5), while higher concentrations of the substrate induce a positive cooperation (h = 1.5). The presence of citrate affects the S0.5 affinity value, but not the Vmax. The presence of citrate (10 mM) removes the cooperative effect at higher concentrations of the substrate, as h = 1.0. A theoretical Ki for citrate was calculated and equals 1.30 mM. PMID- 10365246 TI - Nucleoid proteins of pea chloroplasts: detection of a protein homologous to ribosomal protein. AB - Basic proteins were isolated from purified pea chloroplast nucleoids by acid extraction. Using RP-HPLC, the component composition of the basic proteins was studied. SDS-PAGE of major HPLC-fractions showed that the basic nucleoid proteins are heterogeneous with mol. masses of components from 17 to 30 kDa. One polypeptide with mol. mass of 28 kDa (P28) was obtained by RP-HPLC. The sequencing of three tryptic peptides of P28 (T6, T17, and T19) showed that they are homologous to the ribosomal protein L19 of Saccharomyces cerevisiae. The possible functional role of ribosomal proteins in chloroplast nucleoids is discussed. PMID- 10365247 TI - Mitochondrial changes during the apoptotic process of HeLa cells exposed to cisplatin. AB - HeLa cells undergo apoptosis after exposure to cisplatin. Since mitochondria have recently been proposed as a probable effector of this type of cell death, we performed an analysis using the fluorescent cation rhodamine 123, which is transported actively by this organelle. Cisplatin induces a decrease in the mitochondrial staining, as assessed by cytofluorometric analysis. Microscopic analysis demonstrated that this effect was accompanied by damage of the mitochondria. These features were not exclusive of cisplatin, as other antineoplasic agents (taxol, etoposide) elicited similar effects. These results point toward the notion of a general effect of antineoplasic drugs over the mitochondria during induction of apoptotic cell death. PMID- 10365248 TI - Categorizing reactivity of bacteriorhodopsin cysteine mutants crosslinking to 4 bromoretinal. AB - The structure of bacteriorhodopsin (bR) has been probed by a large number of experimental methods. In earlier work distance constraints measured from the 1BRD Brookhaven structure (1, 2) were used to guide site-directed mutagenesis/affinity labeling experiments (3-5). In the present study we report on the use of limited molecular dynamics (MD) investigations of the same bR/affinity label system. We show here that the chiral center introduced when 4-bromo-all-trans retinal is synthesized produces variable impact on potential crosslinking. Our MD analysis suggests the following ranking of binding site mutants in order of reactivity: R118C > S118C >> S121C > R141C >> S141C >>> R121C, R138C, S138C. Chirality appears to have limited effect for the M118C mutants but shows more dramatic impact for the T121C and S141C mutants. These results are in excellent agreement with the experimental observations and offer encouragement that MD can be a useful component of experimental design with considerable predictive power. PMID- 10365249 TI - Induction of apoptosis by selenite and selenodiglutathione in HL-60 cells: correlation with cytotoxicity. AB - Effects of selenite and selenodiglutathione, an initial metabolite of selenite, on the induction of apoptosis and cytotoxicity were investigated in human promyelocytic leukemia HL-60 cells. Treatment of selenite or selenodiglutathione resulted in concentration-dependent cytotoxicity, measured by lactate dehydrogenase leakage assay, and by tetrazolium salt reduction assay. Selenodiglutathione has been shown to exert more cytotoxic effect than selenite in both assay systems. Time-course study of cellular selenium uptake suggests that the higher cytotoxicity of selenodiglutathione be largely due to faster and greater selenium uptake rate. Treatment with selenite or selenodiglutathione also induced apoptosis in a dose-dependent manner, as detected by enzyme-linked immunosorbent assay and by DNA fragmentation assay. The dose-response data of apoptosis induced by selenite or selenodiglutathione were similar to those of cytotoxicity, implicating a relationship between the induction of apoptosis and cytotoxicity. Zn, which is a well-known inhibitor of apoptosis, dose-dependently blocked not only the induction of apoptosis, but also the membrane damage induced by selenium, corroborating this hypothesis. It was noted that the inhibition of apoptosis by Zn exerted little protective effect on cytotoxicity at higher concentrations of selenium, compared with a perfect protective effect at low concentration of selenium. These results suggest that cytotoxicity induced by selenium may be partially correlated with apoptosis. PMID- 10365250 TI - Inhibition of eukaryotic dna polymerase alpha by persimmon (Diospyros kaki) extract and related polyphenols. AB - The effects of persimmon extract (Diospyros kaki) and related polyphenols on eukaryotic DNA polymerase alpha were examined. It was found that persimmon extract, epigallocatechin gallate, and epicatechin gallate strongly inhibited the activity of DNA polymerase alpha purified from calf thymus. Among these polyphenols, persimmon extract had the most potent effect on DNA polymerase alpha activity and the concentration of persimmon extract producing 50% inhibition of the activity was 0.191 microM. Persimmon extract showed a weaker effect on DNA polymerase beta and slightly inhibited primase and DNA polymerase I. The inhibition of DNA polymerase alpha by persimmon extract was competitive with the template-primer and noncompetitive with dTTP substrate. The Ki value of DNA polymerase alpha for persimmon extract was estimated to be 70 nM. Moreover, persimmon extract inhibited [3H]thymidine incorporation of human peripheral lymphocyte cells stimulated by PHA. PMID- 10365251 TI - Cloning, sequencing, expression and characterization of the manganese superoxide dismutase gene from Vibrio alginolyticus. AB - The sodA gene coding for manganese superoxide dismutase from the marine microorganism Vibrio alginolyticus was cloned, sequenced and over-expressed in Escherichia coli using the pET20b (+) expression vector. The full-length gene was consisted of 603bp open reading frame, which encoded a polypeptide of 201 amino acid residues, with a calculated molecular weight of 22672Da. The deduced amino acid sequence of the sodA showed considerable homology to other Mn-SODs. The recombinant enzyme was efficiently purified from crude E. coli cell lysate by the metal ion affinity chromatography. The recombinant VAMn-SOD resisted thermo denaturation up to 60 degrees C and was insensitive to inhibitors such as H2O2, NaN3 and diethyldithiocarbamic acid. PMID- 10365252 TI - Effect of alpha lipoic acid amide on hexachlorobenzene porphyria. AB - The aim of this work is to study the effect of thioctamide--the commercial form of alpha lipoic acid amide--on the porphyrinogenic action of hexachlorobenzene (HCB). For this purpose, porphyria was induced in rats by chronic HCB treatment, with or without simultaneous thioctamide administration. Two different groups of rats were used as reference: one treated with vehicle (control) and the other treated with thioctamide (TO). Urine delta aminolevulic acid, porphobilinogen, and porphyrin excretions were lower in the HCB + TO treated group than in the HCB group, and the same happened with liver uroporphyrin accumulation. On the other hand, the second stage of uroporphyrinogen-decarboxylase activity was significantly higher in the HCB + TO group than in the HCB group. delta aminolevulic acid synthase activity was higher in the HCB group. Hepatic thiobarbituric acid reactive substances were lower in HCB + TO group than in HCB group. Thus, we might suggest that TO would decrease HCB effects by means of its free radical scavenging ability, and by having a direct effect on uroporphyrinogen-decarboxylase activity. PMID- 10365253 TI - Chemical modification studies on the glucose/mannose specific lectins from field and lablab beans. AB - Seeds of Dolichos lablab var. lignosus (field beans) and variety typicus (lablab beans) contain glucose/mannose specific lectins that have been affinity purified and well characterised (Siva Kumar N., and Rajagopal Rao, D., J.Biosci., 1986, 10, 95-109, (1) Rajasekhar et al., (Biochem.Archives. 1997, 13, 233-240) (2). Purified lectins are glycoproteins with a native molecular mass of 60 kDa and are made of two types of subunits (Gowda et al., 1994, J.Biol.Chem. 269, 18789-18793) (3). Chemical modifications of various groups in purified lectins (using group specific reagents) such as lysine (citraconic anhydride), carboxyl groups (water soluble carbodiimide) tyrosine (N-acetyl imidazole) and tryptophan (2-hydroxy 5 nitro benzylbromide) revealed that 14 out of 21 residues of lysines 7 out of 92 residues of carboxyl groups, 16 out of 24 tyrosine residues and 2 out of 10 tryptophan residues were modified. Lysine and carboxyl group modification led to 95% loss in haemaglutinating activity compared to control while tyrosine and tryptophan modifications led to complete loss of lectin activity. Arginine and histidine modifications led to only 50% loss in activity. The extent of modification and loss in activity was same when the lysine and carboxyl groups were modified in the presence and absence of the inhibitory sugar methyl alpha-D glucopyranoside at 0.1 M concentration. However protection of modification and lectin activity was observed when the tyrosine and tryptophan residues were modified in the presence of the inhibitory sugar. Earlier CD studies carried out (1) and extensive chemical modification studies reported here substantiate the involvement of tyrosine and tryptophan residues in the sugar binding site of these lectins. PMID- 10365254 TI - Fatty acid profile of Escherichia coli during the heat-shock response. AB - The possible changes in the fatty acid profile of Escharichia coli during heat shock have been investigated. Bacteria growing in steady-state at 30 degrees C were subjected to an abrupt temperature upshift to 45 degrees C and held at the high temperature for various periods of time in order to elicit the heat-shock response. Fatty acid compositions of lipids extracted from samples taken at different times after the temperature upshift, as well as from cultures in steady state at 30 and 45 degrees C, were determined by gas-chromatography. It has been found that the total unsaturates to total saturates ratio decreases gradually during heat-shock and that 30 min after the temperature jump, the reduction is equivalent to 57% of the difference between ratios corresponding to steady-state cultures at 30 and 45 degrees C. Consistent with this remodeling of lipid acyl chains, there is a decrease in the excimerization rate of the fluidity probe dipyrenylpropane incorporated into sonicated E. coli lipid extracts. Such modifications occur within the time-span of the heat-shock response, as judged from our previous measurements of the kinetics of change in heat-shock proteins induction ratio. Together, these results indicate that the control of membrane fluidity during the heat-shock response can be accounted for, at least in part, by an important change in the fatty acid composition of Escherichia coli lipids. PMID- 10365255 TI - Can serum amyloid A or macrophage colony stimulating factor serve as marker of amyloid formation process? AB - Amyloid formation depends on amyloid precursor production and is influenced by the activity of the underlying disorder and mediated by some proinflammatory cytokines. In this pilot study we tried to find some specific markers that could establish the activity of the disease. We investigated 45 samples of sera and 38 samples of urine from patients (pts) with secondary amyloidosis (AA), primary amyloidosis (AL), systemic autoimmune diseases with renal impairment (Vasc) and healthy controls (Co). Pts with AA had increased plasma levels of TNF alpha (9.97 +/- 4.22 vs. 2.63 +/- 1.34 pg/mL, p < 0.001) and SAA (43.14 +/- 16.0 vs. 3.42 +/- 0.7 ng/mL, p < 0.05) in comparison with Co. Plasma levels of M-CSF in the AA group were significantly increased in comparison with Co (1077.34 +/- 238.6 vs. 137.71 +/- 19.6, pg/mL, p < 0.001) and also in comparison with Vasc (482.24 +/- 86.7 pg/mL, p < 0.05). Urinary excretions of TNF alpha (8.92 +/- 8.1 vs. 0.17 +/- 0.11 microgram/mol creatinine, p < 0.01), sIL-6R (1.39 +/- 1.14 vs. 0.07 +/- 0.05 g/mol creatinine, p < 0.01) and M-CSF (650.2 +/- 153.7 vs. 33.3 +/- 8.6 micrograms/mol creatinine, p < 0.01) in AA were significantly increased in comparison with Co. Pts with AL had increased plasma levels of M-CSF (819.83 +/- 264.2 vs. 137.71 +/- 19.6 pg/mL, p < 0.05) and urinary excretion of M-CSF (865.0 +/- 188.4 vs. 33.3 +/- 8.6 micrograms/mol creatinine, p < 0.01) in comparison with Co. SAA has a low specificity for amyloidosis but is a sensitive acute phase reactant. TNF alpha, a proinflammatory cytokine, may reflect the activity of the underlying diseases in secondary amyloidosis. M-CSF was increased both in plasma and urine in amyloidosis groups and seems to be the most promising (possibly specific) marker of amyloidosis. PMID- 10365257 TI - Regulation of an 8-kDa peptide involved in testosterone production by luteinizing hormone in rat Leydig cells. AB - Results of Western blot analysis carried out with an interstitial cell extract from male guinea pig and ovarian extract from immature female rats administered equine chorionic gonadotropin (eCG) provide supportive evidence to our earlier suggestion that an 8-kDa peptide is involved in acquisition of steroidogenic capacity by the rat Leydig cells. It was found that though the signal was observed in other tissues such as liver, kidney and lung which do not produce gonadal hormones, the peptide was modulated only by lutenizing hormone (LH) in the rat Leydig cells. PMID- 10365256 TI - Changes in membrane-bound leucine aminopeptidase activity during maturation and ageing of brain. AB - Aminopeptidases are believed to be enzymes that regulate the activity of various neuropeptides. However, their physiological role, as well as their mechanisms of regulation, are not well understood. To analyze a part of the regulatory mechanisms that control the activity of these enzymes, the subcellular distribution of membrane-bound leucyl aminopeptidase activity was studied in rat brain during development and ageing. Except in fetuses, the enzymic activity was greatest in the microsomal fraction in all ages tested. Except in microsomal and myelin fractions, compared with fetuses, leucyl aminopeptidase activity showed a decrease in 1-week-old rats and a subsequent increase to adult levels in 1-month old rats. This profile differed in the microsomal fraction, where the activity increased steadily up to 1-month-old rats. After this age, the activity decreased progressively in 5-month and 24-month-old rats. These results may reflect changes in the functional status of the susceptible substrates during development and ageing. PMID- 10365258 TI - Effect of dose-rate and dose fractionation on radiation-induced hemolysis of human erythrocytes. AB - Human erythrocytes suspended in an isotonic Na-phosphate buffer, pH 7.4 (hematocrit 2%) were exposed under air to gamma radiation at a dose rates of 2.2 kGy.h-1 and 4.2 kGy.h-1. The dose-response curves for hemolysis of erythrocytes indicated that the process of hemolysis is inversely related to the dose-rate. At both dose-rates we observed a reduced level of hemolysis, when erythrocytes were irradiated with a split dose (0.4 kGy + 2.3 kGy with an interval time between the subsequent exposures from 1 to 4 h) in comparison with the same single dose (2.7 kGy). The maximal effect of fractionation was observed when the interfraction time was equal to 3.5 h. The influence of the interfraction temperature on this effect was observed. The results obtained indicate that enucleated human erythrocytes under suitable radiation conditions are capable of repairing radiation damage which leads to hemolysis. PMID- 10365259 TI - The involvement of hexokinases in trehalose synthesis. AB - Yeast cells harboring a MAL2-8c gene accumulate trehalose during the transition phase of growth on glucose due to the presence of the ADPG-dependent trehalose 6 phosphate synthase. Under these conditions, glucokinase appeared not to provide G 6-P for trehalose synthesis and the two hexokinases seemed to act synergistically. After incubation in d-xylose, trehalose levels in these cells dropped almost in 90%, confirming the involvement of both hexokinases in the accumulation of this carbohydrate. Nevertheless, G-6-P levels appeared to be similar in all strains. Some explanations for this paradox are discussed. In stationary phase, neither of the three isoenzymes were involved in trehalose synthesis. Possibly, gluconeogenesis provides the substrate for trehalose synthesis at that stage. PMID- 10365260 TI - Human anti-DNA autoantibodies and induced antibodies against ROS-modified-DNA show similar antigenic binding characteristics. AB - Alterations in DNA structure by hydroxyl radical modification was characterized by UV spectroscopy, Tm, nuclease S1 digestibility and base modification. In view of indicted role of oxygen free radicals in human diseases, an attempt has been made to precisely compare the antigen binding properties of induced antibodies against hydroxyl radical modified DNA with those of naturally occurring anti-DNA autoantibodies. Antibodies induced against ROS-DNA showed diverse antigen binding characteristics which were comparable with those derived from SLE patients. The immune IgG recognized native DNA, heat denatured DNA, and synthetic polynucleotides in B-/B-like conformations. IgG isolated from SLE sera showed preference for ROS-DNA in competition-inhibition assay. The antigenic diversity of induced antibodies and preference of circulating anti-DNA autoantibodies for ROS-DNA over that of native DNA demonstrates the possible role of modified DNA antigens in the pathogenesis of SLE. PMID- 10365262 TI - Purification and characterization of cytochrome C from camel muscle. AB - Cytochrome C was purified from camel skeletal muscles using ammonium sulphate fractionation and gel filtration on Sephadex G25 and Sephecryl S200 columns. The preparations were pure by SDS-PAGE criteria, with molecular weight of 12,300 Dalton. The electrophoretic mobility of camel cytochrome C was the same as that of the horse heart. The spectral characteristics of the isolated cytochrome C were also investigated. PMID- 10365261 TI - Cloning and expression of human cDNA encoding human homologue of pituitary tumor transforming gene. AB - Recently, a potent transforming gene which was exclusively expressed in rat pituitary tumor but not in normal pituitary had been isolated and named as pituitary tumor transforming gene (PTTG). A cDNA clone encoding human homologue of rat PTTG was isolated from human fetal liver cDNA library. It contained an open reading frame of 603 base pairs predicting a protein composed of 201 amino acids with a calculated molecular weight of 26 kDa. The deduced protein showed about 85% homology (78% identity, 7% favored substitution) with the rat PTTG. Northern blot analysis showed that the cDNA hybridized to 1.0 kb mRNA species which was expressed in fetal liver and several cancer cell lines. These results suggest that the presence of the human homologue of rat PTTG gene may not be restricted to pituitary tumor. PMID- 10365264 TI - Indications for pneumonectomy. Pneumonectomy for malignant disease. AB - The anatomic extent of a pulmonary malignancy usually dictates the need for pneumonectomy to achieve a complete resection. The requirement for a pneumonectomy can frequently be predicted by accurate clinical staging, but may also be required due to intraoperative findings relating to tumor invasion or nodal spread. Completion pneumonectomy is usually reserved for locally recurrent lung cancer or early to late complications following pulmonary resection. Malignant involvement of the carina frequently requires sleeve pneumonectomy. PMID- 10365263 TI - A history of pneumonectomy. AB - Successful pneumonectomy in humans was preceded by the development of techniques for lobectomy. If experience in the animal laboratory had not been ignored, fears regarding sudden occlusion of the pulmonary artery and the fate of the postpneumonectomy pleural space would have been allayed. Precise hilar dissection was facilitated by the development of endotracheal anesthesia, which had been used successfully in animal experiments for at least half a century. PMID- 10365265 TI - Indications for pneumonectomy. Pneumonectomy for benign disease. AB - A wide variety of nonmalignant diseases of the lung require pneumonectomy. Pneumonectomy for inflammatory lung disease is frequently associated with high morbidity rates, and the frequencies of postpneumonectomy space empyema and bronchopleural fistula are high. It is essential to treat underlying infections prior to surgery in an effort to minimize the sputum production, maximize the patient's nutritional status, minimize the chance for intraoperative spillage, and decrease the risk of postoperative bronchopleural fistulas and postpneumonectomy space empyemas. Despite the challenges of performing a pneumonectomy for inflammatory diseases, cure rates for MDR-TB, MOTT infections, and fungal disease, including invasive fungal disease, are excellent. Pneumonectomy for trauma is associated with very high mortality, and efforts should be made to avoid pneumonectomy if possible. Pneumonectomy for other benign conditions is unusual. PMID- 10365266 TI - Indications for pneumonectomy. Extrapleural pneumonectomy. AB - Surgical resection is considered a mainstay for the treatment of malignant pleural mesothelioma (MPM), but the indications for extrapleural pneumonectomy in this disease remain controversial. In general terms, an operation contributes to cancer management if it can be performed with low morbidity and mortality and improves local control, overall survival, or quality of life. The potential role of extrapleural pneumonectomy in MPM is best considered in this context. PMID- 10365267 TI - Assessment of operative risk for pneumonectomy. AB - The preoperative evaluation of patients who are candidates for pneumonectomy is valuable in assessing an individual's relative risk of morbidity, permits the patient to make an informed decision regarding surgery with those risks in mind, and helps the treating physicians by alerting them to likely complications. The workup is accomplished with few and relatively inexpensive tests that reliably predict which patients are at greatest risk for pulmonary, cardiovascular, and other surgical complications and death. The guidelines for evaluation are only suggestions, however, and the assessment of individual patients must be tailored to their specific needs according to the judgment of the treating physician. PMID- 10365268 TI - Techniques of pneumonectomy. Standard pneumonectomy. AB - An understanding of the anatomical, three-dimensional organization of the pulmonary hilum is the foundation necessary for pneumonectomy. The incision type and sequence of control of hilar structure are determined by anatomic position, extent of tumor, and patient safety factors. PMID- 10365269 TI - Techniques of pneumonectomy. Pneumonectomy through an empyema. AB - The practical management of the patient with a destroyed lung in association with a preexisting empyema, based on considerable experiences, is discussed. Control of infection before proceeding with pneumonectomy by adequate drainage of the empyema and control of tuberculosis and pneumonia, particularly on the opposite side, is stressed. Pneumonectomy is undertaken through the empyema and usually in the intrapleural plane. PMID- 10365270 TI - Techniques of pneumonectomy. Pleural pneumonectomy. AB - This article reviews the preoperative assessment and selection of patients for extrapleural pneumonectomy, the surgical technique, and recent data outlining our experience with multimodality therapy for MPM. PMID- 10365272 TI - Techniques of pneumonectomy. Sleeve pneumonectomy. AB - Sleeve pneumonectomy is a technically demanding procedure, the indications of which include non-small bronchogenic tumors extending to the tracheobronchial bifurcation without diseased mediastinal nodes. Right sleeve pneumonectomies are best approached through an ipsilateral thoracotomy in the fifth (or fourth) intercostal space. Median sternotomy for left sleeve pneumonectomy gives outstanding exposure to the tracheobronchial bifurcation, and less incisional discomfort and ventilatory restriction than an ipsilateral thoracotomy. If a tracheobronchial anastomosis is under tension, excessive tracheobronchial and mediastinal dissection and perioperative fluid overload are avoided, then the most common and often fatal early (noncardiogenic pulmonary edema) and late (anastomotic dehiscence) complications are significantly lowered. If these guidelines are respected, this operation generates 5-year survival rates exceeding 40%. PMID- 10365271 TI - Techniques of pneumonectomy. Completion pneumonectomy. AB - Completion pneumonectomy refers to an operation intended to remove what is left of a lung partially resected during previous surgery. Completion pneumonectomy is a technically demanding procedure, which carries an increased operative mortality and morbidity. If the planning and the surgical technique are done meticulously, the good prospect for long-term survival justifies the higher risk. PMID- 10365273 TI - Techniques of pneumonectomy. Video-assisted thoracic surgery pneumonectomy. AB - Thoracoscopic major pulmonary resections such as lobectomies or pneumonectomies are the most difficult operations that can be attempted thoracoscopically, and still have limited routine application in thoracic surgical practice. The precise indications for thoracoscopic pneumonectomy are very rare and have not yet been defined precisely; we limited the procedure only to double tumors, small tumors infiltrating the fissure, and small tumors at the secondary carina not amenable to a bronchoplasty procedure. Although the technique still has very limited applications, the advantages include reduced surgical trauma and consequent minimal postoperative pain, a shortened hospital stay, and a rapid resumption of normal activities which ultimately reduces costs. Wider acceptance, larger series, and a more extensive follow-up will assess the role of thoracoscopic anatomical lung resection in modern thoracic surgical practice. PMID- 10365274 TI - Techniques of pneumonectomy. Drainage after pneumonectomy. AB - After most pneumonectomies, the pleural space can be safely closed without drainage. If a chest tube must be used, a balanced drainage system is recommended. This article specifically addresses some of the controversial issues in the early management of the postpneumonectomy space. It also describes the indications, advantages, and disadvantages of drainage and the methods commonly used for this purpose. PMID- 10365275 TI - Physiologic consequences of pneumonectomy. Consequences on the pulmonary function. AB - The cardiac sequelae following pneumonectomy should be anticipated by a thorough preoperative evaluation of cardiac risk factors, and any identified significant risk should be evaluated and corrected. The most common cardiac complication following pneumonectomy is atrial dysrhythmia. The possible causes, significant correlates, and rationale for prophylaxis are discussed. With a large portion of the pulmonary vascular bed removed by pneumonectomy, the possibility and consequences of right ventricular dysfunction are outlined. Finally, the rare but catastrophic occurrence of cardiac herniation is described. PMID- 10365276 TI - Physiologic consequences of pneumonectomy. Consequences on the pulmonary function. AB - When pneumonectomy is done in children, there is some speculation that lung growth occurs. In the adult population, volume response is incomplete and attributable to alveolar distention rather than multiplication. Stretch is widely regarded as the initial stimulus for compensatory growth. The authors review data pertaining to the physiology of the various adjustments that occur after pneumonectomy gathered both from observations on human response and from experimental findings in animals. Mechanisms and mediators of this adaptive response are discussed. PMID- 10365277 TI - Physiologic consequences of pneumonectomy. Consequences on the esophageal function. AB - Esophageal and upper gastrointestinal dysmotility occur after both pneumonectomy without pulmonary replacement and recipient pneumonectomy for thoracic organ transplantation. After pneumonectomy without pulmonary replacement, there is a shift of the esophagus to the side of pneumonectomy and disturbance of esophageal peristalis. After recipient pneumonectomy for thoracic organ transplantation, esophageal dysmotility and delayed gastric emptying are common. Injury of the vagal nerves, local ischemia, postoperative scarring of the esophagus and mediastinum, and disturbance of the autonomic nervous systems are the major causes of the abnormality. To reduce the incidence of esophageal dysmotility after pneumonectomy, every effort should be made during surgery to prevent direct injury of the esophagus or the vagal nerves. PMID- 10365278 TI - Physiologic consequences of pneumonectomy. Long-term consequences of pneumonectomy done in children. AB - Lung resections in children are performed for a variety of reasons including congenital malformations, infections, bronchiectasis, and tumors. There are no long-term reports on pneumonectomy alone in children, but those on lung resection as a group state that children tolerate these operations well, with mild sequelae if any, and that the majority of them in adulthood can perform non-physically demanding jobs adequately. The authors' findings concur with the reports that younger patients can endure pulmonary resections with minimal functional limitations. PMID- 10365279 TI - Type 2 diabetic subjects without prior myocardial infarction are at the same risk of coronary events as non-diabetic subjects with prior myocardial infarction. PMID- 10365280 TI - From systole to diastole: will the detection of regional diastolic dysfunction allow recognition of coronary artery disease at an earlier stage? PMID- 10365281 TI - Mid-term results of percutaneous mitral balloon valvotomy. PMID- 10365283 TI - Surgery or radiofrequency catheter ablation for atrioventricular nodal reentrant tachycardia--the impossible choice. PMID- 10365282 TI - Specific thrombin inhibitors ... only one tool of the antithrombotic armamentarium. PMID- 10365284 TI - Cardiological aspects of aviation safety. PMID- 10365285 TI - Interventional cardiology in Europe 1995. Working Group Coronary Circulation of the European Society of Cardiology. PMID- 10365286 TI - Regional diastolic function in ischaemic heart disease using pulsed wave Doppler tissue imaging. AB - AIMS: The aim of this study was to determine the utility of pulsed wave Doppler tissue imaging in the evaluation of regional left ventricular diastolic function in patients with ischaemic heart disease. METHODS AND RESULTS: In 30 normal subjects and 43 patients with ischaemic heart disease, Doppler tissue imaging was performed in each of the 16 segments of the myocardium. The following diastolic pulsed wave Doppler tissue imaging parameters were obtained for each segment: (1) regional early diastolic peak velocity (regional e wave cm.s-1); (2) regional late diastolic peak velocity (regional a wave cm.s-1); (3) regional diastolic e/a velocity ratio; and (4) the regional isovolumic relaxation time, defined as the time interval from the second heart sound to the onset of the diastolic E wave. In patients with ischaemic heart disease, each of these parameters was evaluated and compared in ischaemic and normally perfused segments, based on the presence or absence of obstructive lesions of the supplying coronary artery. In patients with coronary artery disease, several differences were observed between diseased and normal wall segments: the mean segmental peak early diastolic velocity (e wave) was reduced (mean +/- SD: 6.4 +/- 2.1 cm.s-1 vs 8.5 +/- 2.8 cm.s-1; P < 0.01); the e/a diastolic velocity ratio was decreased (0.95 +/- 0.3 vs 1.5 +/- 0.6, respectively; P < 0.01) and the regional isovolumic relaxation time was prolonged (104 +/- 36.7 ms vs 69.6 +/- 30 ms; P < 0.01. No differences were observed in any of these parameters between the normally perfused segments of ischaemic patients and normal subjects. Patients with a normal transmitral diastolic Doppler inflow pattern had a mean of 3.7 +/- 2.7 myocardial segments with a local e/a pulsed wave Doppler tissue imaging velocity ratio < 1, fewer than those with an inverted diastolic transmitral Doppler inflow pattern (10.3 +/ 3 segments; P < 0.001). Overall sensitivity and specificity for an inverted local e/a ratio and a local isovolumetric relaxation time > or = 85 ms were of 62% and 72% and 69% and 80%, respectively. CONCLUSION: Regional diastolic wall motion is impaired at baseline in ischaemic myocardial segments, even when systolic contraction is preserved. Pulsed wave Doppler tissue imaging is a useful non-invasive technique which allows the assessment of regional diastolic performance and dynamics of the left ventricular myocardium. Further studies are required to define this role in the evaluation of coronary heart disease. PMID- 10365287 TI - The effect of a low molecular mass thrombin inhibitor, inogatran, and heparin on thrombin generation and fibrin turnover in patients with unstable coronary artery disease. AB - AIM: This study evaluated a novel specific thrombin inhibitor, inogatran, in comparison with unfractionated heparin, with regard to markers for coagulation activity in patients with unstable coronary artery disease. METHODS AND RESULTS: In the Thrombin Inhibition In Myocardial Ischaemia (TRIM) study patients were randomized to one of three different doses of inogatran or to unfractionated heparin, given intravenously over 72 h. In a subpopulation of 320 patients, markers for coagulation activity were measured at baseline, during and after the study infusion. Prothrombin fragment 1 + 2, indicating thrombin generation, decreased in the low, medium and high dose inogatran groups and in the heparin group during the first 6 h of treatment by 12%, 15%, 21% and 26%, respectively. From 6 h to 72 h after the start of infusion the levels changed by -7%, -6%, -4% and +34%, respectively. The increase in the heparin group continued after the infusion was stopped. Thrombin-antithrombin complex, also indicating thrombin generation, decreased by 0%, 2%, 18% and 22%, respectively, during the first 6 h of treatment. During the same period soluble fibrin, an intermediate in fibrin formation, increased both in the low and medium inogatran group by 9%, while a decrease by 4% and 18%, respectively, was seen in the high dose inogatran group and in the heparin group. Fibrin dissolution, as measured by fibrin D-dimer, decreased during the first 24 h of treatment by 20%, 18%, 18% and 20%, respectively. The first 24 h after discontinuation of infusion, fibrin D-dimer increased by 6%, 23%, 25% and 44%, respectively. After 72 h, at the end of infusion, patients treated with inogatran, to a larger extent than those given heparin, had suffered from death, myocardial infarction or refractory angina pectoris. After 7 days this trend was less marked. CONCLUSION: The more pronounced decrease in thrombin generation and fibrin turnover during the first 6 h of infusion, and the later increase in thrombin generation and fibrin turnover, in the heparin group, as compared to the inogatran groups, may be related to the lower clinical event rate during infusion with heparin compared with inogatran and the recurrence of ischaemic events, early after cessation of heparin infusion. PMID- 10365288 TI - Predictors of clinical events or restenosis during follow-up after percutaneous mitral balloon valvotomy. AB - AIMS: The purpose of this study is to define predictors of events or restenosis during follow-up after percutaneous mitral balloon valvotomy. METHODS AND RESULTS: Percutaneous mitral balloon valvotomy was attempted in 137 patients with severe mitral valve stenosis. In 127 patients follow-up was complete with a mean of 4.2 +/- 2.6 years. Events during follow-up were defined as death, mitral valve surgery or repeat percutaneous mitral balloon valvotomy. Restenosis was defined as a decrease in mitral valve area from > or = 1.5 cm2 following percutaneous mitral balloon valvotomy to < 1.5 cm2. There was 80 +/- 4% event-free survival 4 years after percutaneous mitral balloon valvotomy. Multivariate analysis showed chronic atrial fibrillation at baseline (P = 0.039, relative risk (RR) = 2.5) and a high residual maximal gradient after percutaneous mitral balloon valvotomy (P = 0.004, RR = 2.0 per 5 mmHg) to be independent predictors of an event during follow-up. The restenosis rate was 28.3% after 4 years. Chronic atrial fibrillation at baseline (P = 0.0338, RR = 2.2), a small mitral valve area after percutaneous mitral balloon valvotomy (P = 0.0003, RR = 0.8/0.1 cm2) and a high residual maximal transmitral gradient (P = 0.0252, RR = 1.6/5 mmHg) were all independent predictors of restenosis. CONCLUSION: Patients with chronic atrial fibrillation and a high maximal transmitral gradient after percutaneous mitral balloon valvotomy have a higher risk for events during follow-up. Restenosis is related to the presence of chronic atrial fibrillation at baseline and a suboptimal percutaneous mitral balloon valvotomy result. PMID- 10365289 TI - Comparison of late results of surgical or radiofrequency catheter modification of the atrioventricular node for atrioventricular nodal reentrant tachycardia. AB - AIMS: Although arrhythmia surgery and radiofrequency catheter ablation to cure atrioventricular nodal reentrant tachycardia differ in technical concept, the late results of both methods, in terms of elimination of the arrhythmogenic substrate and procedure-related new and different arrhythmias, have never been compared. This constituted the purpose of this prospective follow-up study. METHODS AND RESULTS: Between 1988 and 1992, 26 patients were surgically treated using perinodal dissection or 'skeletonization', and from 1991 up to 1995, 120 patients underwent radiofrequency modification of the atrioventricular node for atrioventricular nodal reentrant tachycardia. The acute success rates of surgery and radiofrequency catheter ablation were 96% and 92%, respectively. Late recurrence, rate in the surgical and radiofrequency catheter ablation groups was 12% and 17%, respectively. Mean follow-up was 53 months in the surgical group and 28 months in the radiofrequency catheter ablation group. The final success rate after repeat intervention was 100% in the surgical group and 98% in the radiofrequency catheter ablation group. Comparison of the initial and recent series of radiofrequency catheter ablated patients showed an increased initial success rate with fewer applications. In the radiofrequency catheter ablation group, a second- or third-degree block developed in three patients (2%), requiring permanent pacing, whereas in the surgical group no complete atrioventricular block was observed. Inappropriate sinus tachycardia needing drug treatment was observed in 13 patients (11%), mostly after fast pathway ablation, but was never observed after surgery. New and different supraventricular tachyarrhythmias arose in 27% of the patients in the surgical group and in 11% of the radiofrequency catheter ablation group, but did not clearly differ. CONCLUSION: This one-institutional follow-up study demonstrated comparable initial and late success rates as well as incidence of new and different supraventricular arrhythmias following arrhythmia surgery and radiofrequency catheter ablation for atrioventricular nodal reentrant tachycardia. Today radiofrequency catheter ablation has replaced arrhythmia surgery for various reasons, but the late arrhythmic side-effects warrant refinement of technique. PMID- 10365290 TI - Influence of ambulance crew's length of experience on the outcome of out-of hospital cardiac arrest. AB - AIMS: To investigate whether an ambulance crew's length of experience affected the outcome of out-of-hospital cardiac arrest. METHODS AND RESULTS: This was a population-based, retrospective observational study of attempted resuscitations in 1547 consecutive arrests of cardiac aetiology by Nottinghamshire Emergency Ambulance Service crew. One thousand and seventy-one patients were managed by either a paramedic or a technician crew without assistance from other trained individuals at the scene of arrest. Overall, the chances of a patient surviving to be discharged from hospital alive did not appear to be affected by the paramedic's length of experience (among survivors, 18 months experience vs non survivors 16 months experience, P = 0.347) but there appears to be a trend in the effect of a technician's length of experience on survival (among survivors, 60 months experience vs non-survivors 28 months experience, P = 0.075). However, when a technician had 4 years of experience or more and a paramedic 1 year's experience, survival rates did improve. Logistic regression analysis, adjusted for factors known to influence outcome, revealed that chances of survival increased once technicians had over 4 years of experience after qualification (odds ratio 2.71, 95% CI 1.17 to 6.32, P = 0.02) and paramedics after just 1 year of experience (odds ratio 2.68, 95% CI 1.05 to 6.82, P = 0.04). CONCLUSIONS: Survival from out-of-hospital cardiac arrest varies with the type of ambulance crew and length of experience after qualification. Experience in the field seems important as paramedics achieve better survival rates after just 1 year's experience, while technicians need to have more than 4 years' experience to improve survival. PMID- 10365291 TI - Transcatheter closure of secundum atrial septal defects with the new self centering Amplatzer Septal Occluder. AB - AIMS: The study was set up to find out whether a new self-centering prosthesis for transcatheter closure of secundum atrial septal defects could overcome the disadvantages of previously described devices. METHODS AND RESULTS: Fifty-two consecutive patients with a significant atrial septal defect were considered for transcatheter closure with the Amplatzer Septal Occluder. The device, made of a Nitinol and polyester fabric mesh, provides a different approach to defect occlusion by stenting the atrial septal defect up to a stretched diameter of 26 mm. Three infants whose large defects were demonstrated on a transthoracic echocardiogram were excluded from transcatheter treatment. On transoesophageal echocardiography, 49 defects ranged from 6-26 mm, in one adult the defect measured 28 mm and this patient was excluded from attempted transcatheter closure. At cardiac catheterization in five further patients, devices were not implanted, in two because the stretched diameter exceeded 26 mm and in three the device was withdrawn because it was unstable or compromised the mitral valve. Thus, device closure was performed in 43 patients. At follow-up after 3 months the complete closure rate was 97%. CONCLUSION: The self-centering Amplatzer Septal Occluder is very efficient and user-friendly and offers interventional closure in 83% of an unselected group of patients presented with an atrial septal defect. PMID- 10365292 TI - Screening for familial hypertrophic cardiomyopathy using brain natriuretic peptide. PMID- 10365293 TI - Therapeutic options in the management of articular contractures in haemophiliacs. AB - Haemophilic contracture is seen most commonly as an equinus deformity of the ankle, or at the knee or elbow in the form of a flexion deformity. Treatment options are varied, and decision-making is based on the degree of the contracture, its chronicity, the presence of articular subluxation, the patient's ability to participate in treatment, and the available medical facilities. The treatments available fall into four categories: physiotherapy, orthotics, corrective devices, and surgical procedures. Treatment should be primarily by physiotherapy, splintage, and corrective devices. The late or severe case may require surgical correction in the form of soft-tissue procedures. Soft-tissue correction of muscle shorthening may be performed such as lengthening of the Achilles tendon for equinus deformity of the ankle, or hamstring release of the flexor muscles of the knee. Lower femoral osteotomy has been used for correction of flexion deformity at the knee joint. Mechanical distraction using external fixators for treatment of severe knee flexion contractures has been recently reported with satisfactory results. The main principle underlying the treatment of haemophilic contracture is the restoration of the patient's lifestyle and mobility, rather than anatomic or radiographic normality. PMID- 10365294 TI - Physiotherapy for the prevention of articular contraction in haemophilia. AB - The idea of prevention and preventative care is not new. History and culture have given us many examples of the importance of physical well-being and the prvention of illness and disease. The ancient societies of China focused on the balance of Yin and Yang in promoting health; Greece and Rome valued the importance of health and physical culture; the earliest Hebrew societies documented the importance of diet and dietary restrictions as a means towards good health. Through this century health professionals have advocated the importance of preventive care as an integral element of the quality of health. Haemophilia is a life-long condition with a high potential towards disability, handicap and impairment if not adequately treated. It is therefore essential that those with haemophilia are taught the importance of physical fitness at an early age as a means of preventing articular contractures. Physiotherapy is of great importance in this field, especially in third-world countries where the supply of replacement products are scarce or non-existent. PMID- 10365295 TI - Physiotherapy for the treatment of articular contractures in haemophilia. AB - Articular contractures in haemophilia are impairments that can not be cured by means of physiotherapy because of the pathophysiology of the joint. Rehabilitation, however, tries to diminish the disabilities and prevent handicaps caused by the impairments. Physiotherapy aims at pain reduction by means of manual traction. Next to manual traction the intensive physiotherapy programme includes mobilization techniques, muscle strengthening exercises and stretching, joint stability training, postural and gait, training, and functional training. In all 50 haemophilia patients have undergone this intensive 4-week clinical rehabilitation programme. Data of 20 of these severe haemophilia patients show that the mean range of motion at the start of the rehabilitation period, after 4 weeks and after 5 years do not differ. In spite of progressing arthropathy after 5 years the activities of daily living (ADL), walking range and pain are equal or better according to 13 of 15 patients. PMID- 10365296 TI - Non-operative treatment of flexion contracture of the knee in haemophilia. AB - For the non-operative treatment of flexion contracture of the haemophilic knee we have used serial casting and wedging in 58 patients, and extension/de-subluxation orthoses in 13 patients. On average it was possible to achieve -5 degrees of extension by 4 weeks, with only a little improvement in the following 4 weeks. The short--to medium-term results using either the extension/de-subluxation hinges or serial casting were similar. Both methods have been shown to result in significant improvement in joint contracture. PMID- 10365297 TI - Hamstring release and posterior capsulotomy for fixed knee flexion contracture in haemophiliacs. AB - Between 1977 and 1981 hamstring release (n = 27) and posterior capsulotomy (n = 22) were performed on 27 haemophilic patients. The follow-up of 19 patients is documented for at least 5 years after the operation. The mean age was 30 years (15-40 years), the mean follow-up period was 12.5 years (5-20 years). Postoperatively 16 of 19 knee joints achieved immediate full extension, three patients showed 10 degrees extension deficiency. In the long run the original good result could be demonstrated in just 11 patients. Using the Orthopedic Advisory Committee of the World Federation of Hemophilia (OAC) Scores there were three good, 12 satisfactory and four poor results. In minor forms of arthropathy a hamstring release with posterior capsulotomy is the only recommended operation. In severe forms we would actually recommend a combination with other procedures such as arthroscopic synovectomy or joint replacement. PMID- 10365298 TI - Extensor supracondylar femoral osteotomy as treatment for flexed haemophilic knee. AB - The experience with extensor supracondylar femoral osteotomy as treatment for the flexed haemophilic knee is presented with the description of 19 patients treated during a 30-year period (1968-98). The average age of the patients was 16 (8-35 years), and the average age follow-up was 13 years (3-30 years). Six patients had flexion fixed deformity while the rest presented 40 degrees average range of motion (10-75 degrees). In 13 patients a single osteotomy without internal fixation was performed, in one an osteosynthesis with a condylar plate and in five stabilization was achieved by means of Blount staples. Previous surgery was performed in two patients with patello-femoral ankylosis. The osteotomy site was consolidated in every patient and the deformity was corrected. Two bleeding complications were observed: one haemarthrosis and one psoas haematoma. Flexion relapsed in one patient who underwent another procedure after 12 years. One patient presented with a peroneal nerve paralysis; another one a genu recurvartum; which required flexor osteotomy. The extensor supracondylar femoral osteotomy is a procedure that aligns the limb with scarce modification of articular mobility. PMID- 10365299 TI - Correction of fixed contractures during total knee arthroplasty in haemophiliacs. AB - Encouraging clinical experience has increased the indications for prosthetic knee arthroplasty in haemophiliacs. The medical status, physical disability, age and projected activity levels are the major factors in determining treatment for the patient with unilateral or bilateral haemophilic arthropathy of the knee. In more severely involved knees of patients with haemophilia, flexion contracture is a common deformity. In addition, valgus, external rotation deformity and posterior subluxation of the tibia may exist. The surgeon must have the expertise and experience to correct these deformities sufficiently when performing a total knee arthroplasty. A properly performed soft-tissue release which achieves balance between the medial and lateral ligamentous structures and posterior capsule can provide stability to the knee with a semiconstrained prosthesis. In cases with severe deformity requiring resection of the posterior cruciate ligament a posterior cruciate substituting prosthesis may be necessary. PMID- 10365300 TI - Management of fixed flexion contracture of the elbow in haemophilia. AB - The authors stress that prevention of flexion contractures and artropathy by early factor replacement and physical therapy for every haemophiliac is the standard of care. Physical therapy, serial casting, and Quengel cast correction have not proven successful in correction of fixed flexion contractures at the elbow. In the patient who has a flexion contracture that interfered with function, an attempt at physical therpay combined with the use of either the Dynasplint or Flowtron will be tried. If there is no response, a surgical synovectomy combined with a possible radial head resection and anterior capsular release would be the authors' procedure of choice. In the face of advanced arthropathy, the authors would consider a distraction arthroplasty. PMID- 10365301 TI - The role of orthoses in the management of elbow joints in persons with haemophilia. AB - The elbow joint is extremely prone to haemarthroses, and when these occur the elbow requires support and rest. Supporting the limb with a simple collar-and cuff sling, a triangular bandage either with or without a shoulder immobilization device will provide an analgesic effect without adding an unnecessary heavy orthosis. As the swelling subsides and the pain lessens a dynamic orthosis aids in the support and compression. When considering functional branching of the elbow joints there are a number of basic principles to be followed. One requires to consider problems of orthosis suspension, two dimensional motion and stabilization in the functional range of motion. This article provides the rationale for the special needs of persons with haemophilia who suffer from involvement of their elbow joints. PMID- 10365302 TI - Management of equinus contractures of the ankle in haemophilia. AB - Equinus deformity has been a significant problem in haemophilia. It causes difficulties in walking and secondary problems in adjacent joints. There are a number of potential causes in haemophilia. A careful history, examination, and plain radiographs will determine the aetiology, which frequently is multifactorial. Hopefully, prophylactic factor replacement will reduce the incidence of such problems in the future. Prompt 'on demand' therapy will reduce the complications of articular and soft-tissue bleeds. Physiotherapy, splints, and orthotics will usually allow a full recovery of function and comfort. Rarely, surgical intervention is required to correct articular and/or musculo-tendinous problems. The choice of surgery depends upon the cause(s) of the deformity and should only be undertaken in experienced haemophilia units following careful counselling of the patient regarding the aims and nature of the operation and the patient's involvement in an effective rehabilitation programme. PMID- 10365303 TI - Common orthopaedic problems in haemophilia. AB - The most important fact in patients with haemophilia is the avoidance of recurrent haemarthroses by means of haematological prophylaxis. Unfortunately this is not currently possible in the majority of countries around the world. When only on-demand haematological treatment is available, frequent evaluations are necessary for the early diagnosis and treatment of intra-articular bleeding episodes. The typical outcome of these patients is towards the development of chronic synovitis, and later on to haemophilic arthropathy, if the state of synovitis is not promptly and adequately controlled. Once arthropathy develops the functional prognosis is poor, although haemophilic patients use to tolerate very well their tremendous joints destructions. Treatment of these patients should be performed in a comprehensive basis within a multidisciplinary haemophilia unit. PMID- 10365304 TI - Postal stamps on drug and alcohol abuse. PMID- 10365305 TI - Role of caspases in apoptosis and disease. AB - Apoptosis, a genetically governed process of eliminating cells in response to a variety of stimuli provides protection against cancer and viral infections as well as maintains homeostasis. Recent studies using both molecular and cloning approaches, and in vitro systems have identified a class of highly specific proteases, termed caspases, that appear to have an important role in apoptotic execution. Caspases are synthesized as precursor molecules that require processing at specific aspartate residues to produce the active enzyme which in turn leads to the cleavage of various death substrates that lead to morphological changes typical of apoptosis. This review discusses caspases, their inhibitors and regulators. Since cytotoxic drugs used in chemotherapy of leukemia's and solid tumors cause apoptosis in target cells, elucidating the consequences of proteolytic activity occupies a central role for understanding of the molecular mechanism of apoptosis which can help us to use the caspase inhibitors as targets of therapy. PMID- 10365306 TI - Pharmacology of sildenafil citrate. AB - Erectile dysfunction is a common and multi-factorial disease that strongly impairs the quality of life in men. During the past decade, many new therapeutic strategies have become available. But the need for oral treatment was strongly felt. This need appears to have been fulfilled with the introduction of sildenafil. The drug acts by enhancing smooth muscle relaxant effect of nitric oxide. A number of clinical studies have now proved its safety and efficacy. The drug has shaken social life all over the world and to accept this "magic pill" or not remains the question of individual choice. PMID- 10365307 TI - Immunohistological localisation of vascular endothelial growth factor in human endometrium. AB - Several polypeptide growth factors regulate epithelial and stromal development in endometrium under the influence of estrogen and progesterone, and thereby regulate growth and differentiation of endometrium during menstrual cycle. However, little is known about the angiogenic growth factors that may affect endometrial vasculature throughout each menstrual cycle. Vascular endothelial growth factor (VEGF) is suggestively an important angiogenic growth factor in the female reproductive tract. The aim of the present study was to immunolocalize and assess semi-quantitatively VEGF immunostaining in cells of proliferative phase (n = 3), secretory phase (n = 6) and hyperplastic (n = 6) human endometrial samples. VEGF concentrations were significantly higher in glandular (P < 0.001) and stromal (P < 0.01) compartments of proliferative stage endometrium compared with those in secretory stage and hyperplastic endometrial samples, with no difference in the scores for glandular and stromal compartments between secretory stage and hyperplastic endometrial samples. Generally, glandular expression of VEGF was higher as compared to stromal compartment. Thus, it appears that endometrial VEGF expression and concentration are enhanced by estrogen, and may be correlated with neovascularization and increased vascular permeability during late proliferative period. Additionally, there was no enhancement in VEGF expression in hyperplastic glands, suggesting that regulation of glandular growth and that of angiogenesis in human endometrium operate through different mechanisms. PMID- 10365308 TI - Role of Ca2+ on uterine force stimulated by a glycoside from the root of Dalbergia saxatilis. AB - Uterine muscle contraction is dependent on external Ca2+ and Ca2+ release from cytoplasmic storage sites. In this study, the mechanism of Ca2+ mobilization in uterine muscle cells by glycoside, dalsaxini, isolated from the root of D. Saxatilis was investigated in the rat. Uterine muscle contractility stimulated by dalsaxin was concentration dependent (ED50 0.13 mg/ml) and was significantly attenuated (85%; P < 0.01) in Ca(2+)-free physiological solution and in solutions containing verapamil (0.06-0.48 mumol). The small transient contraction observed in Ca(2+)-free medium was further suppressed by caffeine (2 mmol) and completely abolished in solutions containing Lanthanum chloride [(La3+), 2 mmol]. Contractions stimulated by the glycoside were unaffected by amiloride (50-83 mumol) in Ca(2+)-free and Ca(2+)-containing media. Dalsaxin also altered the pattern of uterine contraction stimulated by high potassium depolarization from fast-phasic to a sustained but transient plateau. It is concluded that dalsaxin causes uterine muscle contraction by mobilizing external Ca2+ through predominantly a voltage-dependent Ca2+ channel. PMID- 10365309 TI - Body fat topography in Indian and Tibetan males of low and normal body mass index. AB - Body fat topography was determined using anthropometric techniques in young, healthy, Indian and Tibetan adults. Indian subjects had significantly higher fat contents with greater abdominal obesity when compared with Tibetans matched for body mass index (BMI). This differential fat distribution may contribute, in part, to the greater cardiovascular risk of Indians. Using a cross sectional model, the data was also analysed to assess the probable changes in body fat topography with weight gain. This model suggests a preferential gain in abdominal subcutaneous fat as compared to other sites. This data may have implications while evaluating disease risks with weight gain. PMID- 10365310 TI - The development and validation of a digital peak respiratory pressure monitor and its characteristics in healthy human subjects. AB - A digital peak respiratory pressure (DPRP) monitor for determining maximum inspiratory pressure (MIP) and maximum expiratory pressure (MEP) was developed using a pressure transducer and an analog to digital converter. It was calibrated using a mercury manometer. Human studies were conducted in healthy young adults in order to determine within-subject and inter-individual variability, as well as diurnal variations and gender differences in maximal respiratory pressures. The calibration studies for the instrument indicated that the instrument recorded accurate pressures, with little temporal drift. Within-subject variability was generally low while inter-individual variability was higher and significant. Gender differences were similar to those recorded in literature for other racial groups. The DPRP monitor described is inexpensive, accurate and portable, making it ideal for use at the patient's bedside. PMID- 10365311 TI - Thyroid functions in pre-eclampsia and its correlation with maternal age, parity, severity of blood pressure and serum albumin. AB - Maternal thyroid function was investigated in 32 pre-eclamptic women and 10 normal pregnant women in their third trimester. Serum total tri-iodothyronine (TT3) and total thyroxine (TT4) were decreased significantly (P < 0.001) and TSH was increased significantly (P < .001) in pre-eclampsia as compared to normal pregnancy. There was no influence of parity and maternal age on thyroid functions. TT3 and TT4 decreased significantly (P < .001) with increase of serum albumin, while there was no correlation of TT4 with serum albumin. PMID- 10365312 TI - Independent and combined effects of L-arginine and diazepam on ammonium chloride induced convulsions in rats. AB - The independent and combined effects of L-arginine (840 mg/kg) and diazepam (0.75 mg/kg) pretreatment (30 min) were tested on ammonium chloride (400 mg/kg)-induced convulsions in rats. Ammonia concentrations were determined in blood and brain regions (cerebral cortex, brain stem and cerebellum) 30 min after L-arginine or diazepam treatment. Ammonia concentrations were measured at the time of induction of convulsions by ammonium chloride in L-arginine, diazepam or saline pretreated animals. L-arginine and not diazepam decreased ammonia concentrations in control as well as in ammonium chloride-treated animals. However, both the compounds suppressed convulsions elicited by ammonium chloride. Protection produced concurrently by these agents was much greater than that produced by them independently. It is concluded that convulsions caused by hyperammonemic condition can be suppressed either by preventing a rise in brain ammonia to toxic level or by anticonvulsant agents having a GABA potentiating action. A much greater protection can be achieved if agents having these properties are administered concurrently. PMID- 10365313 TI - Brainstem auditory evoked potentials among rubber factory workers. AB - The study was conducted on 27 rubber factory workers for the functional assessment of brainstem auditory pathway. Neurobehavioural questionnaire was administered to the workers and the personal sampler was used to evaluate the respirable particulate load inhaled per day of each worker along with qualitative analysis for PAH compounds. Evoked potential recording was carried out for brainstem auditory responses. Chest X-rays of workers exhibited varied abnormal features. Multiple regression analysis of data showed definite prolongation of latencies with increasing concentration of respirable particulate load though it was not statistically significant. Comparison with normative data indicated prolongation of latencies of rubber factory workers. PMID- 10365314 TI - Role of estradiol in the capacitation and acrosome reaction of hamster epididymal spermatozoa in the isolated uterus of mice incubated in vitro. AB - The site of sperm capacitation, the agents and mechanisms causing capacitation and acrosome reaction (AR) in vivo are not well understood. The female reproductive tract has been reported to play a key role during capacitation and AR. Some experiments were carried out on the capacitation and AR of hamster epididymal spermatozoa in the estrogen and progesterone dominated uterus (estrous and diestrous respectively) albino mice, incubated in TALP without calcium and BSA. Also the effect of estradiol (200 micrograms/ml) supplemented to TALP, on capacitation and AR was examined. Capacitation and AR of hamster spermatozoa incubated in the isolated uterus of both estrous and diestrous mice were significantly (P < 0.05) higher in the presence of exogenous estradiol than that in its absence. Acrosome shedding occurred earlier i.e. at 3rd hour as compared to the in vitro studies where it occurred at 5th hour. The present study thus reveals that uterus of both estrogen and progesterone dominated mice play an important role in the induction of capacitation and AR. The addition of estradiol might have the influence on the synthesis of uterine proteins of mice which might be important for capacitation and AR. PMID- 10365315 TI - Stress due to exams in medical students--role of yoga. AB - A student under optimal stress does bring out his or her best, However extremes of stress can result in stress induced disorders and deteriorating performance. Can yoga be of benefit in stress induced effects in medical students? The present study was conducted in first MBBS students (n = 50) to determine the benefit if any of yogic practices on anxiety status during routine activities and prior to examination. Feedback scores were assessed to determine how the students had benefited from the practices. Anxiety status as assessed by Spillberger's anxiety scale showed a statistically significant reduction following practice. In addition the anxiety score which rose prior to exams showed a statistically significant reduction on the day of exam after practice. These results point to the beneficial role of yoga in not only causing reduction in basal anxiety level but also attenuating the increase in anxiety score in stressful state such as exams. The results of the exam indicated a statistically significant reduction in number of failures in yoga group as compared to the control group. The improvement in various parameters such as better sense of well being, feeling of relaxation, improved concentration, self confidence, improved efficiency, good interpersonal relationship, increased attentiveness, lowered irritability levels, and an optimistic outlook in life were some of the beneficial effects enjoyed by the yoga group indicated by feedback score. PMID- 10365316 TI - Factors influencing changes in tweezer dexterity scores following yoga training. AB - Yoga has already been shown to improve perceptual-motor skills, but the factors which influence its effects are not well defined. This study correlates age, gender, and motivation to learn yoga with the performance in a dexterity task following yoga. Tweezer dexterity was recorded in eighty subjects belonging to four groups. Two groups were given a month of yoga training. One group consisted of subjects who had volunteered to join for the training and the other group were deputed for the training as part of their job. The two remaining groups did not receive yoga training and were selected to match the respective groups receiving yoga, for age and sex, but not for their motivation to learn yoga. The test involved using a tweezer to place metal pins in evenly spaced holes in a metal plate within four minutes. Following yoga the scores of the volunteers who learnt yoga increased significantly, whereas there was no change in scores of deputed subjects and non-yoga groups. For reasons described in detail, factors such as age and gender did not appear to contribute to the difference in performance. Hence motivation to learn yoga appeared to influence the magnitude of increase. PMID- 10365317 TI - Effect of Hepatogard--an indigenous formulation on dexamethasone induced antihealing effects in male albino rats. AB - Hepatogard, is a multi-ingredient phytopharmaceutical product containing crude powders of eleven plants. Its effect on the different parameters of wound healing was assessed alone and in the presence of dexamethasone. The parameters chosen for the study were the breaking strength of incised wound, breaking strength of granulation tissue and hydroxyproline content. The result showed that Hepatogard increased the breaking strength of granulation tissue but not of incised wound. It reversed the dexamethasone induced decrease in breaking strength in both incised wound and granulation tissue. Even though it had no effect of its own on hydroxyproline concentration, it reversed the dexamethasone induced decrease in the hydroxyproline content of granulation tissue. Thus, Hepatogard has the potential for antagonizing the antihealing effect of steroids in patients receiving steroid therapy. PMID- 10365318 TI - Evaluation of antiallergic activity (type I hypersensitivity) of Inula racemosa in rats. AB - Alcoholic extract of root of Inula racemosa, was studied for its antiallergic effect in experimental models of type I hypersensitivity, viz. egg albumin induced passive cutaneous anaphylaxis (PCA) and mast cell degranulation in albino rats. The alcoholic extract was prepared by the process of continuous heat extraction. LD50 of this extract was found to be 2100 +/- 60 mg/kg, i.p. Assessment of protection against egg albumin induced passive cutaneous anaphylaxix by different doses of Inula racemosa was done by giving drug intraperitoneally or orally for seven days or once only. Mast cell degranulation studies were done by using compound 48/80 as degranulation agent with same dosage schedule. Inula racemosa (i.p. as well as p.o.) showed significant protection against egg albumin induced PCA. Protection against compound 48/80 induced mast cell degranulation by alcoholic extract of Inula racemosa (single dose) was similar to that of disodium cromoglycate. The seven days drug treatment schedule showed greater protection than disodium cromoglycate intraperitoneally. The results suggest that Inula racemosa possesses potent antiallergic properties in rats. PMID- 10365319 TI - Objective structured practical examination in pharmacology for medical laboratory technicians. AB - The objective structured practical examination (OSPE) is a useful evaluation method for testing psychomotor skills. Students for the degree in medical laboratory technology require to learn certain skills which will make them useful in any research or teaching laboratory in experimental pharmacology. We outline an OSPE which can be used for evaluating students in experimental pharmacology. PMID- 10365320 TI - Gentamicin induced inhibition of steroidogenic enzymes in rat testis. AB - Gentamicin is an aminoglycoside antibiotic, widely used for treating many gram negative bacterial infections. Though nephrotoxicity is the most highlighted side effect, it has also been found to cause an alteration in the phosphatase activities of testes and accessory sex organs and a decline in the sperm count. This study was designed to assess the effects of gentamicin on testicular steroidogenesis and to ascertain whether such alterations are reversible. Laboratory inbred adult, male, 'Wistar' strain rats were chosen as the experimental animal. A significant dose-dependant reduction in the activities of the two steroidogenic enzymes, accompanied with a significant decrease in ascorbic acid and elevation of level of cholesterol was observed. The effects were maximum at a dose of 100 mg/kg, b.wt. After 15 days of withdrawal of the drug therapy the biochemical parameters namely ascorbic acid and cholesterol returned to normal levels whereas the activities of the two dehydrogenases showed a compensatory increase. This indicates that gentamicin affects the steroidogenic enzymes, causing an alteration in the formation of testosterone, which was manifested in the elevated cholesterol in the adult rat testes. However, these alterations were reversible. PMID- 10365321 TI - Pharmacokinetics of phenytoin: unaltered by enalapril and amlodipine in rhesus monkeys. AB - A cross over single and multiple dose study was carried out to find out pharmacokinetic interactions between diphynylhydantoin (DPH) (35 mg/kg, p.o.) and antihypertensives enalapril (1.6 mg/kg; p.o.) and amlodipine (0.4 mg/kg, p.o.) in rhesus monkeys. Neither the plasma concentrations nor the pharmacokinetic parameters of DPH were altered by coadministration of enalapril or amlodipine, suggesting that enalapril and amlodipine can be safely administered to epileptic patients receiving phenytoin. PMID- 10365322 TI - Comparative study of beta-adrenoceptor blocking efficacy of two formulations of esmolol in vivo and in vitro. AB - The objective of the present study was to compare the cardiovascular beta blocking activity of two different formulations of esmolol. Spontaneously beating guinea-pig isolated atria and the heart rate and blood pressure of anaesthetized cat were employed in the study to compare the beta-blocking efficacy of the two formulations of esmolol using isoprenaline as an agonist. In guinea-pig isolated atria the standard esmolol formulation (Brevibloc) reduced basal atrial rate more significantly than the indigenously formulated esmolol (test formulation). Both the formulations produced similar parallel rightward shift of cumulative concentration response curves of isoprenaline with closely comparable pA2 values. In anaesthetized cats, only indigenous esmolol formulation significantly decreased basal heart rate. Both the formulations did not modify the basal blood pressure and isoprenaline-induced fall in blood pressure, despite significantly blocking isoprenaline-induced tachycardia. It is suggested that both the formulations produced similar degree of beta-1 adrenoceptor blocking activity. PMID- 10365323 TI - Evaluation of anti-tumor efficacy of injectable Centchroman in mice bearing Ehrlich ascites carcinoma. AB - Anti-tumor efficacy of Centchroman formulated as niosomes and gel implant was evaluated in Swiss albino mice bearing Ehrlich ascites carcinoma at 10 mg/kg body weight dose given subcutaneously. Median day of death, percentage increase in host life span and changes in body weight were studied. Centchroman significantly (P < 0.05) increased the median day of death both in free and formulated systems. Also, injectable formulations exhibited a significant (P < 0.05) increase in host life span compared to free drug, hence, enhanced anti-tumor efficacy against Ehrlich ascites carcinoma. PMID- 10365324 TI - Noise in the operation theatre: intensity and sources. PMID- 10365325 TI - Osmotic fragility of normal and sickle haemoglobin containing red blood cells. PMID- 10365326 TI - [The ocular surface]. PMID- 10365327 TI - [Concentric changes in the visual field associated with GABA-mimetic antiepileptic agents]. AB - Bilateral visual field constriction has been recently reported in patients treated with vigabatrin. It has been considered that vigabatrin, a GABA agonist antiepileptic drug, was specifically responsible for this visual field defect. We present four observations sharing the same characteristics of chronic tunnel vision. Three patients had had vigabatrin but the fourth one received other antiepileptic drugs, progabide, an agonist of post-synaptic GABA receptors, and phenobarbital which interferes with GABA-A receptors. It is thus possible to hypothesize a retinal toxicity triggered by chronically increased GABA transmission. If this is confirmed, an accurate incidence of symptomatic and asymptomatic visual field constriction with GABA-mimetic drugs should be established, as well as the patients' profiles which are more at risk. Patients currently under this type of treatment should be checked by both manual and automatic perimetry every six months to one year. PMID- 10365328 TI - [Treatment of ocular cicatricial pemphigoid with sulfasalazine]. AB - PURPOSE: To report on three patients with biopsy-proven ocular cicatricial pemphigoid successfully treated with sulphasalazine. METHODS: Three case reports. RESULTS: A 71-year-old man, treated with dapsone for ocular cicatricial pemphigoid stopped his treatment because of an allergy to this drug. Oral sulphasalazine, 2.5 grams daily was successfully used as an alternative treatment (3 month follow-up). Two patients, aged 71 and 84 year old, were treated with dapsone for ocular cicatricial pemphigoid. Both patients stopped their treatment because of drug induced hemolytic anemia. They then received oral sulphasalazine, 4 grams daily. The disease was successfully controlled. In the first patient, sulphasalazine was discontinued after 13 months; and in the second patient no relapse was seen after a 16 month follow-up period. No adverse side effect of sulphasalazine occurred. CONCLUSION: Sulphasalazine, that has already been proven to be effective for Crohn's disease, also can be used in ocular cicatricial pemphigoid. However, further studies including a larger series of patients along with a longer follow-up are necessary to confirm the efficacy of sulphasalazine in this disease. PMID- 10365329 TI - [Peribulbar anesthesia for peroperative and postoperative pain control in eye enucleation or evisceration: 31 cases]. AB - OBJECTIVES: The aim of this prospective study was to assess peroperative and postoperative analgesia in eye enucleation or evisceration performed under peribulbar anesthesia. PATIENTS AND METHODS: We report 31 patients undergoing an eye enucleation (17 cases) or evisceration (14 cases). The surgical procedure was performed under local anesthesia alone in 22 patients. General anesthesia was associated with local anesthesia in 9 patients. Peribulbar block was achieved with the first insertion of the needle parallel to the inferior orbital floor and the second at level of supraorbital notch. A mixed anesthetic solution of equal quantity of lidocaine 2% with epinephrine (0.25 mg/20 ml) and bupivacaine 0.50% with epinephrine (0.10 mg/20 ml) was injected (total quantity 16.8 +/- 4.3 ml). RESULTS: To assess the peroperative pain we considered the patients with local anesthesia only (22 patients). One of these 22 patients needed one injection (0.50 mg/kg) of propofol for cutting the optic nerve. Surgery was ended without any other drug but that case was considered as a failure. Peroperative analgesia was obtained in 21 of 22 patients (95.4%). To assess analgesia in the postoperative period we included 31 patients. Analgesia was complete from the accomplishment of the peribulbar block to the 4th hour in all patients (efficacy 100%). From the 4th to the 24th hour, pain remained absent in 11 (enucleation 10 cases and evisceration 1 case) of the 31 patients and no drug was used. In 20 patients (enucleation 7 cases and evisceration 13 cases), pain appeared between the 4th and the 10th hour and patients were relieved by paracetamol alone in 14 cases (enucleation 6 cases and evisceration 8 cases) or by its association with nalbuphine in 5 cases (enucleation 1 case and evisceration 4 cases). In one patient (evisceration) the association of the drugs was uneffective. CONCLUSION: Peribulbar anesthesia is safe and generates major postoperative analgesia so we suggest to offer that technique to patients undergoing evisceration or enucleation. PMID- 10365330 TI - [Visual acuity and ocular diseases in aged residents of nursing homes: study of 219 persons in Orleans]. AB - PURPOSE: To measure the visual acuity and to determine the etiological causes of visual impairment in the elderly residing in nursing homes. METHODS: 219 elderly persons residing in nursing homes were examined in the residence. The ophthalmological examination consisted in a visual acuity measurement, a slit lamp examination and a fundus examination. RESULTS: This study included 145 women and 74 men. Mean age was 79.1 years (range 41-101 years). Visual acuity could be measured in 181 subjects (82.6%): it was 1/10 or worse in the better eye in 23 of them (13%) and 2/10 to 3/10 in 36 patients (20%). In 21 (17.6%) out of the 119 patients aged over 74 years, visual acuity was 1/10 or worse in the better eye. Visual impairment significantly increased with age (p < 0.05). There was no difference between men and women in the prevalence of visual impairment. Among the 55 subjects with visual impairment, the main causes of vision loss were: cataract in 36 patients (66%), age-related macular degeneration in 9 patients (16%) and optic neuropathies in 5 patients (9%). Only one (2.8%) out of the 36 patients with cataract could be operated. CONCLUSION: The rate of visual impairment of people in nursing homes was higher than in corresponding age groups in the general population. The main cause of vision loss was cataract; however, only a few patients could benefit from an operation. These results confirmed that a systematic ophthalmologic examination should be performed before general health problems prevent patients from being operated. PMID- 10365331 TI - [Intraorbital dermoid cyst. Apropos of a case]. AB - Orbital dermoid cyst represents 3 to 4% of all primary orbital tumours. The intraorbital location is relatively rare. We report the case of a 9 year-old male child with an orbital dermoid cyst, who presented with progressive right proptosis, which had developed 3 months earlier. Visual acuity, ocular motility and funduscopic examination were normal. Imaging aspects (ultrasonography and computed tomography) have revealed an extraconical orbital, hypodense, well limited tumor, without bone destruction and no enhancement located at the postero external side of the orbit. The tumors was extirpated surgically via a superior approach. Histopathology confirmed the diagnosis of dermoid cyst. PMID- 10365332 TI - [Main semeiologic characteristics of ptosis]. AB - BACKGROUND: We report the main characteristics of blepharoptosis. MATERIALS AND METHODS: All cases of blepharoptosis treated at the Reims University Hospital from 1992 to 1997 were reviewed. Ptosis, levator function, palpebral aperture, and the position of the upper crease were recorded as well as results of the epinephrine test, acetylcholine esterase inhibitor antibodies and computed tomography findings. RESULTS: There were 96 unilateral and 34 cases of bilateral blepharoptosis (164 cases). The cases of congenital blepharoptosis (36 cases) were usually unilateral with severe ptosis and poor levator function. In cases with Claude Bernard Horner syndrome, the blepharoptosis was unilateral with minimal ptosis and a positive response to neosynephrine. In cases with oculomotor nerve palsy (31 cases) the ptosis was moderate to severe and levator function was poor. In those with myasthenia, the prostigmine test was positive. Levator aponeurotic disinsertion was observed in 16 cases with severe ptosis, positive response to neosynephrine and a high upper crease. There were three cases of trauma-induced ptosis, 5 cases of myopathy and 26 cases of idiopathic ptosis. DISCUSSION: The clinical presentation of blepharoptosis is usually related to the etiology. Bilateral blepharoptosis is most often observed in congenital forms, levator disinsertion or idiopathic cases. Male sex predominates in congenital ptosis and oculomotor palsy, and female sex in Claude Bernard Horner syndrome. Moderate to severe ptosis is observed in congenital ptosis or oculomotor palsy. The upper crease is absent in many cases of congenital blepharoptosis and is high in case of levator disinsertion. The response to neosynephrine is positive in Claude Bernard Horner syndrome and aponeurotic disinsertion. The response to prostigmine is positive in case of myasthenia. CONCLUSIONS: Clinical aspects of blepharoptosis are related to etiology. The different features guide surgical or medical treatment of blepharoptosis. PMID- 10365333 TI - [NO donors and experimental retinal vein occlusion]. AB - PURPOSE: The development of extended territories of nonperfused capillaries after branch vein occlusion (b.v.o.) is correlated to the secondary constriction of the arteriole crossing the occluded territory. Local NO release is impaired soon after b.v.o. and accounts for the secondary arteriolar constriction. In this report we present evidences showing that administration of an NO donor can reverse the secondary arteriolar vasoconstriction observed after b.v.o. MATERIAL AND METHODS: Simultaneous preretinal NO profiles and arteriolar diameter measurements were performed in miniature pigs after experimental b.v.o. The effect of preretinal microinjections of the NO-donor Sodium Nitroprusside on the arteriolar diameter was studied. RESULTS: A significant arteriolar vasoconstriction occurring in parallel with a pre-retinal [NO] decrease was observed 4 hours after b.v.o. Microinjection of the NO-donor SNP caused a segmental, reversible arteriolar dilatation. CONCLUSION: The present results, suggest that local NO supply in the first hours following b.v.o. may contribute to protect the retina against ischemic injury. PMID- 10365334 TI - [Acquired enophthalmos associated with hypoplasia of the maxillary sinus and asymptomatic chronic maxillary sinusitis]. AB - BACKGROUND: We report a case of progressive enophthalmos, maxillary sinus hypoplasia and radiological signs of sinusitis in a young woman. CASE REPORT: The patient had no history of sinus surgery or facial trauma. There were no signs of sinusitis. Functional endoscopic sinus surgery was proposed to restore the communication between the nose and the antrum. DISCUSS: The particular aspects of asymptomatic maxillary sinus and causes of maxillary sinus hypoplasia are discussed. We suggest that obstruction of the osteomeatal complex causes negative pressure in the maxillary sinus provoking bone alterations. These alterations cause the maxillary sinus hypoplasia and the enophthalmos. PMID- 10365335 TI - [Results of suture fixation in esotropia with non-accommodative far-near uncomitance]. AB - PURPOSE: In esotropia, the dynamic component is the unique objective of suture fixation. The aim of this study was to assess outcome after the suture fixation used with or without other conventional methods and to determine the value of the anesthesia sign for evaluating the innervational factor of these esotropias. METHODS: Unilateral and bilateral posterior fixation sutures were applied in 54 cases exhibiting far-near incomitance between 10-20. The same procedure was applied in 14 cases with similar type of esotropia with incomitance greater than 20. RESULTS: A 100% success rate was achieved when suture fixation was used alone. When suture fixation was applied in combination with other conventional methods for cases with a static and a dynamic component the success rate was 86% (37/43 cases) in unilateral myopexy and 92% (12/13 cases) in bilateral myopexy. DISCUSSION: In cases with this type of esotropia, suture fixation, the only means of correcting a dynamic angle, gives good results as the component disclosing the apparent angle is well-documented. In addition, these findings emphasize the value of the general anesthesia sign to determine the innervational factor causing the dynamic angle. PMID- 10365336 TI - [Clinical examination of the lacrimal film]. PMID- 10365337 TI - [Ocular rosacea]. PMID- 10365338 TI - [Clinical exploration of the lipid phase of the laryngeal film using Tearscdope plus]. PMID- 10365339 TI - [Legislative aspects of the corneal graft]. PMID- 10365340 TI - [Materials for intraocular lenses. II. Silicone intraocular lenses]. PMID- 10365341 TI - [Surgical technique of limbal autotransplantation in severe and recent eye burns]. AB - We report our surgical technique of limbal autograft for recent severe ocular burns. Our procedure is a modified Kenyon and Tseng technique. We discuss our main observations when using this procedure for serious ocular burns which demonstrate the physical signs best indicating surgery, the limbal region to be used, and optimal postoperative follow-up. PMID- 10365342 TI - [Lacrimal sac cyst: apropos of a case]. AB - A 17-year-old girl presented a unilateral cyst of the lacrimal sac. Epiphora was induced by compression of the patent lacrimal duct. The bony nasolacrimal canal was enlarged on the affected side. At pathology examination, the epithelium of the cyst wall was identical to the sac epithelium. PMID- 10365343 TI - Fetal defaecation: is it a normal physiological process? AB - It has been long known that the late gestation human fetus passes meconium in response to hypoxia. However, there is good evidence, from amniotic fluid studies measuring bile pigment and enteric enzyme content, to suggest that passage of meconium is a normal physiological event in the second trimester. Similarly there is some indirect evidence that fetal defaecation is a normal physiological process in the third trimester. However, this evidence is less strong, and it is safer to assume that in most cases meconium staining of liquor at this time is associated with fetal hypoxia. Dilation of the rectosigmoid portion of the distal bowel found in newborn infants with anorectal malformations supports the hypothesis that fetal colonic peristalsis and defaecation is a normal physiological process. PMID- 10365344 TI - Does postoperative ventilation have an effect on the integrity of the anastomosis in repaired oesophageal atresia? AB - Several authors have claimed that the use of postoperative ventilation or graded withdrawal of respiratory support reduces the incidence of anastomotic complications after repair of oesophageal atresia, particularly where the gap between the oesophageal ends has been extensive or where the anastomosis has been constructed under tension. Careful review of their data reveals little objective evidence to either support or refute this contention. Many institutions are achieving low leakage rates following oesophageal anastomosis in oesophageal atresia, but to date there has been no controlled study to show that the use of neck flexion, muscle paralysis, intubation and assisted ventilation postoperatively influences the integrity of the anastomosis. The sequence of observations that led to the presumed relationship between postoperative ventilation and oesophageal leak is reviewed. It would appear that the effect of postoperative ventilation and paralysis on the oesophageal anastomosis is yet to be determined. PMID- 10365345 TI - Clinical research in aboriginal health. PMID- 10365346 TI - Pancreatic enzyme replacement therapy in cystic fibrosis: Australian guidelines. Pediatric Gastroenterological Society and the Dietitians Association of Australia. AB - Pancreatic enzyme replacement therapy (PERT) is a major factor associated with achieving optimum growth and nutritional status in cystic fibrosis (CF) patients with pancreatic insufficiency and consequent malabsorption. Currently in Australian CF clinics policies for the usage of PERT vary considerably. This paper highlights the current issues related to fat absorption and use of PERT in CF. It also provides evidence to support the recommendation of a PERT dose based on a standard ratio of lipase units per gram of dietary fat consumed for individual patients. A consistent approach to PERT doses will facilitate identification of patients whose PERT intake increases their risk of fibrosing colonopathy and for whom gastroenterological review is warranted. Recent reports indicate that PERT intake can be reduced with a secondary improvement in growth and nutrition status as a consequence of increased dietetic input. These Australian guidelines for the judicious use of PERT in CF should lead not only to a refinement in nutritional management of patients with CF but should also facilitate an improvement in compliance with therapy due to sophistication in patient education materials. The Australian guidelines for the use of PERT in CF if correctly applied, will also provide patients and their families with a better understanding of the relationship between PERT and nutritional status. PMID- 10365347 TI - Randomised controlled trials addressing Australian aboriginal health needs: a systematic review of the literature. AB - OBJECTIVE: To describe the frequency and design of controlled clinical trials specifically addressing the health needs of Aboriginal Australians. METHODOLOGY: Electronic searching of Medline, the Australasian Medical Index, the Aboriginal and Torres Strait Islander health bibliographic database, and handsearching of Aboriginal Health: an annotated bibliography. Studies that met the following selection criteria were included: i) addressed an Aboriginal health problem, ii) had a formal description of methods and results, and iii) compared the health effects of an intervention with a concurrent control group. All summary data were extracted by a single author. RESULTS: Only 13 studies were identified. Nine were randomised controlled trials and four were controlled trials but not randomised. Only one of these involved adults, which is unexpected. Although important Aboriginal child health issues were addressed in 12 of the 13 studies, most were undertaken many years ago and may not be familiar to Australian paediatricians. The majority appeared to be designed appropriately. Interestingly, the two studies not published in the medical literature had the largest sample size. There is no evidence that the number of published clinical trials involving Aboriginal Australians is increasing, or that long-term applied clinical research programmes have been established. CONCLUSION: There is a profound lack of well designed studies assessing medical interventions. This is further evidence that Australia has failed to develop a research infrastructure able to inform health care in Aboriginal communities. Adults appear especially disadvantaged. PMID- 10365348 TI - Six-month follow-up of children with obstructive sleep apnoea. AB - OBJECTIVES: This study examined prospectively changes in development, temperament and sleep related behaviour in children referred for obstructive sleep apnoea (OSA) and polysomnographic sleep study, some of whom had surgical intervention. METHODOLOGY: Using a prospective cohort study design, parents of 56 children referred for OSA completed sleep and temperament questionnaires and their child was assessed developmentally at the time of the polysomnographic sleep study. Forty (72%) of the children were neurologically normal. At 6 months, 42 children were reassessed using sleep and temperament questionnaires and a developmental assessment. After excluding the primary snorers, subjects were categorised as having had intervention (n = 24) or not (n = 15), and differences over the 6 month period in Griffiths scores, temperament and sleep related behaviour were examined. RESULTS: Regardless of intervention status, there was an improvement in night-time and day-time sleep behaviour for the total group, though the extent of improvement was more marked in the intervention group. For the neurologically normal children, improvement in the sleep behaviour was only significant for the intervention group (P < 0.05). Intervention did not result in any significant changes in Griffiths developmental score or temperament. CONCLUSION: Surgical intervention improves sleep behaviour in children though not temperament or development. PMID- 10365349 TI - Aetiological factors and development in subjects with obstructive sleep apnoea. AB - OBJECTIVE: To examine whether maternal pregnancy complications, adverse birth events, respiratory illnesses, or developmental difficulty were increased in neurologically normal children with obstructive sleep apnoea (OSA) and whether severity of OSA adversely affects the child's development and temperament. METHODOLOGY: Maternal report of perinatal events, respiratory illness and developmental difficulty in 37 children with OSA was contrasted with a comparison group (n = 67). Children with OSA were assessed developmentally (Griffiths Scales), had a parental rating of temperament (Australian Temperament Scale) and attended an overnight polysomnographic sleep study. RESULTS: Children with OSA had an increased prevalence of adverse maternal pregnancy and perinatal events, respiratory disease and developmental concerns. Limited associations were found between the severity of OSA and development or temperament difficulty. CONCLUSIONS: This study suggests a relationship between OSA, though not its severity, and pre/perinatal adversity and child development. Polysomnographic and detailed developmental assessment of community-based samples of children with OSA and control children are necessary to confirm these findings. PMID- 10365350 TI - Hospital admissions in the first year of life in very preterm infants. AB - OBJECTIVE: To analyse hospital readmissions to 1 year in infants < 33 weeks' gestation. STUDY DESIGN: Cohort of very preterm infants born in Western Australia. METHODS: Parental social class, history of asthma, race, gestational age, birthweight, sex, severity of respiratory disease and oxygen requirement at 28 days chronic lung disease (CLD), 36 weeks and term, maternal smoking, cohabitation with siblings, breast-feeding duration and hospital readmissions were recorded prospectively. RESULTS: Data were available for 538 of 560 (96%) infants discharged. Eight died in the first year. Two hundred and twenty-five infants (42%) had 443 readmissions, of which 370 were medical and 73 surgical. Risk factors for medical readmission were Aboriginal race, male sex and CLD. Breast-feeding was protective. Risk factors for surgical admission were male sex, lower gestation, severe hyaline membrane disease, severe CLD and birthweight < 10th centile. CONCLUSIONS: Readmission is common after very preterm birth. Risk factors for medical and surgical admission differ with CLD being the only perinatal factor associated with both medical and surgical admission. PMID- 10365351 TI - Seroprevalence of Helicobacter pylori infection in Malaysian children: evidence for ethnic differences in childhood. AB - OBJECTIVES: To determine the prevalence of Helicobacter pylori (H. pylori) in healthy Malaysian children and to discover whether differences exist among children of different races. METHODS: Serum samples from asymptomatic children tested for H. pylori seropositivity using an ELISA test. RESULTS: Five hundred and fourteen healthy urban Malaysian children aged 0.5 to 17 (mean 5.9) years from three different racial groups had their blood tested for H. pylori antibodies. The overall prevalence was 10.3%. There was no significant difference in the prevalence of infection between boys and girls, but a significant rise was noted with increasing age (P = 0.009). Seropositivity was most common in the Indians and lowest in the Malays (P = 0.001). Father's level of education did not affect the child's rate of H. pylori seropositivity. CONCLUSION: The prevalence of H. pylori seropositivity among asymptomatic urban Malaysian children is lowest in Malays. Intermediate in Chinese and highest in Indians. The racial differences found in children are consistent with those found in Malaysian adults. PMID- 10365352 TI - An iron treatment trial in an aboriginal community: improving non-adherence. AB - OBJECTIVE: To compare supervised vs unsupervised oral iron treatment in anaemic Aboriginal children living in a remote community with a 40% prevalence of iron deficiency anaemia. METHODOLOGY: A randomised unblinded clinical trial in children < 6 years presenting to a remote Health Centre with anaemia. Oral iron prescribed as a daily unsupervised dose (group A) was compared to twice weekly supervised administration (group B) over 12 weeks. Parenteral iron (group C) was reserved for failure of oral treatment. RESULTS: Only 3 of 25 children in group A responded to treatment compared to 23 of 26 children in group B (odds ratio = 7.7, 95% confidence interval 2.6-25.0). After six weeks of treatment, the mean haemoglobin rise was 0.96 g/L in group A compared to 10.9 g/L in group B and 12.4 g/L in group C. On entry to the study, 29.4% of subjects were underweight, 33.3% stunted and 35.3% microcephalic. The mean catch-up in weight/height on iron treatment over the study was only 0.28 (0.08, 0.48) Z-scores. CONCLUSIONS: Oral iron as directly observed twice weekly treatment is superior to unsupervised therapy. In view of the poor compliance with unsupervised treatment and the high prevalence of iron deficiency anaemia (along with stunting and microcephaly) in Aboriginal children in northern Australia, we propose to undertake in partnership with communities a nutritional intervention program with a high energy weaning food fortified with micronutrients (iron, vitamin A, zinc, folate) as the most effective strategy to address these nutritional problems in the weaning period. PMID- 10365353 TI - Alarm settings for the Marquette 8000 pulse oximeter to prevent hyperoxic and hypoxic episodes. AB - OBJECTIVE: To determine safe and appropriate alarm limits for the Marquette 8000 pulse oximeter to prevent hyperoxic and hypoxic episodes in neonates. It is necessary to define these limits for each brand of oximeter because of the variance in nonuser adjustable calibration algorithms used in pulse oximeters. METHODOLOGY: Oxygen saturation values obtained from a Marquette 8000 pulse oximeter (SpO2) were compared with simultaneous arterial blood gas PaO2 values obtained from blood gas analysis, for 322 samples in 24 consecutive neonates (median 30 weeks' gestation). RESULTS: In order to prevent 95% of hyperoxic episodes (PaO2 > 90 mmHg), the upper alarm limit was 95% SpO2. Similarly, to prevent 95% of hypoxic episodes (PaO2 < 40 mmHg), the lower alarm limit was 95% SpO2. A sensitivity lower than 95% had to be accepted to develop an alarm range which prevented both hyperoxic and hypoxic episodes. To maintain PaO2 values between 40 and 90 mmHg, an appropriate alarm range of 94-97% SpO2 (90% sensitivity, 28% specificity) was established. CONCLUSIONS: The relative merits of high sensitivity versus high specificity should be considered when determining appropriate alarm limits. Alarm limits which represent a balance between sensitivity and specificity will minimise false alarms and provide a clinically practical range. It would be useful for this type of information to be available for each brand of oximeter, to assist the user in determining appropriate alarm settings. PMID- 10365354 TI - Feeding problems in infants with gastro-oesophageal reflux disease: a controlled study. AB - OBJECTIVE: Gastro-oesophageal reflux disease (GORD) in infants is commonly associated with feeding problems but has not been subject to systematic controlled study. We evaluated feeding, dietary, behavioural data obtained from systematic objective studies of six-month old infants with and without GORD. METHODS: Infants with GORD (defined by 24-h pH monitoring, n = 20), and age, gender, gestation, and socio-economic matched healthy infants (n = 20) had standardised assessments of dietary intake, oromotor function by videoanalysis (Feeding Assessment Schedule, FAS), and infant feeding behaviour by Testers and Maternal Ratings (TRIB and MRIB). Videofluoroscopic analyses of swallowing was undertaken in 11/20 GORD infants and analysed by standardised paediatric check list. RESULTS: Compared with control data: GORD infants had significantly lower energy intakes; the FAS showed GORD infants to have significantly fewer adaptive skills and readiness behaviour for solids, significantly more food refusal and food loss; the TRIB showed GORD infants to be significantly more demanding and difficult with feeds; and the MRIB revealed that mothers of GORD infants had significantly more negative feelings, significantly less enjoyment of feeds, and reported significantly more crying behaviour. On videofluoroscopy, oral preparatory and oral phase problems predominated, particularly with solids, silent aspiration occurred during the pharyngeal phase in 2/11, and delayed oesophageal transit occurred in 4/11. CONCLUSIONS: Feeding problems affecting behaviour, swallowing, food intake, and mother-child interaction occur in infants with GORD, who displayed a lack of development of age-appropriate feeding skills. The contribution of feeding problems to morbidity in GORD in infants has been underestimated in the past. PMID- 10365355 TI - Repeated dose inhaled budesonide versus placebo in the treatment of croup. AB - OBJECTIVE: To investigate the efficacy and tolerance of 12-hourly dosing with 2 mg 4 mL-1 of inhaled budesonide versus placebo in patients admitted to hospital with moderate/severe croup. METHOD: Eighty-two children hospitalised with croup received either 2 mg 4 mL-1 of budesonide or placebo 12 hourly (maximum four doses) via Ventstream nebuliser in a randomised, double-blind manner. Croup scores were performed at 0, 2, 6, 12, 24, 36 and 48 h from initial nebulisation whilst the patient remained hospitalised. Follow-up assessments were made 1 and 3 days after discharge. RESULTS: Improvement was observed in the budesonide group over the 12-h dosing interval when compared to placebo (P = 0.04). Time to attain a significant clinical improvement was superior in the budesonide group (P = 0.01). Three days after discharge seven of 32 placebo-treated patients and one of 34 budesonide-treated patients had sought further medical follow-up (P = 0.02). CONCLUSION: Twelve-hourly dosing with inhaled budesonide significantly improved symptoms of croup as well as decreased relapse rates when compared with placebo. PMID- 10365356 TI - Neurodevelopmental outcome of very low birth weight babies admitted to a Malaysian nursery. AB - OBJECTIVE: To examine the prevalence and pattern of neurodevelopmental handicap at 2 years of age in very low birth weight infants (VLBW) admitted in 1993 to a level 3 Malaysian nursery. METHODS: All VLBW babies born in the hospital or referred for neonatal care during 1993 were enrolled prospectively in the study. At 2 years of age development was assessed using the Griffiths mental scales. Neurological, hearing and visual assessments were graded into five groups according to functional handicap. Control infants were randomly selected during attendance at a primary health care clinic. RESULTS: One hundred and fifty VLBW infants were admitted and 82 (54.6%) survived to 2 years, of whom 77 (93.9%) were assessed. The mean General Quotient (GQ) on the Griffiths Scales was 94 (15.7) for the study group and 104 (8.3) for the 60 controls. For GQ, 21 (27.3%) of the study population were 1 or more SD below the mean (18 between 1 and 2 SD and 3 > 2 SD) compared with 1 (1.6%) of the controls who was 1-2 SD below the mean. Visual impairment occurred in 2 study infants and none of the controls. There was no hearing impairment in either group. Cerebral palsy occurred in 3 (1 mild and 2 moderate-severe) of the study group and none of the controls. Functionally 18 (23.3%) of the study group had mild handicap, 1 (1.3%) moderate, 2 (2.5%) severe, 2 (2.5%) multiply severe and 54 (70.2%) were normal. CONCLUSION: Although survival was low, overall rates of functional handicap were similar to those reported in developed countries but the proportion with moderate or severe handicap was low. PMID- 10365357 TI - Cisapride and caesarean section: their role in babies with gastroschisis. AB - OBJECTIVE: The objective of this study was to compare the neonatal postoperative course and morbidity for patients with gastroschisis who received cisapride with those who did not receive cisapride. STUDY DESIGN: Data were obtained by review of the medical records of all the patients with gastroschisis who were admitted to Sydney Children's Hospital between January 1984 and December 1995. Data were compared between 15 babies who received cisapride with 27 who did not. The mode of delivery and outcome of babies in whom gastroschisis was diagnosed antenatally was compared with those who were diagnosed at birth. RESULTS: Duration to the commencement of feeds, attainment of full feeds and the length of hospital stay were not statistically different between these two groups, with or without cisapride (p = > or = 0.1). There were more elective Caesarean sections in the antenatally diagnosed group compared to those detected at birth and the outcome of these two groups showed no statistically significant difference. CONCLUSIONS: Our study identified no benefit from cisapride therapy in babies with gastroschisis and also there was no benefit from elective Caesarean section for babies with antenatal diagnosis of gastroschisis. PMID- 10365358 TI - An evaluation of the autopsy following death in a level IV neonatal intensive care unit. AB - OBJECTIVE: To ascertain the determinants of neonatal autopsy, define clinical errors in the causes of death, and elucidate the possible audit and genetic value of the autopsy following death in a Level IV neonatal intensive care unit (NICU). METHODS: A review and correlation of clinical and autopsy information in a case series of infants who died during the period 1991-97. RESULTS: Two hundred and twenty-nine of 4057 infants admitted to the NICU died and 91 (39.7%) underwent an autopsy. The underlying cause of death was significantly different in infants who had an autopsy compared with infants who did not (P = 0.02). The autopsy rate was higher for deaths from miscellaneous causes (52.9%), lethal malformation (46.8%) and infection (45.4%) than deaths from prematurity (25.9%) and asphyxia (19%). Clinical errors in the causes of death were found in 22% of the infants, and in 4.4% a change in management may have been curative or prolonged life. The autopsy had audit value in 26% of infants and genetic value for a single gene (Mendelian) disorder in 4.4%. CONCLUSIONS: Although the autopsy following death in a Level IV NICU yields potentially useful information in more than one-third of cases, this does not seem sufficient to ensure a high neonatal autopsy rate. PMID- 10365359 TI - Atrial septal defect in Malta. AB - OBJECTIVE: To study diagnostic and surgical trends in atrial septal defect (ASD) in a population-based study, and estimate birth prevalence and spontaneous closure rates. METHODOLOGY: All patients in Malta diagnosed as having ASD and born between 1990 and 1994 were identified from various sources. This took place in the setting of a regional hospital supplying diagnostic services for the entire population. Echocardiographic follow-up was also undertaken for lesions not requiring intervention. RESULTS: A total of 190 patients born in this period were diagnosed as having ASD. Age at diagnosis and age at surgery have decreased significantly over the period under study (P < 0.0001). The mode of diagnosis has become entirely noninvasive, and the perioperative mortality decreased dramatically over time. For the period 1990-94, the incidence at birth for defects not requiring intervention was 2.0/1000 live births, defects requiring intervention 0.4/1000 live births. A total of 92% of 50 defects not requiring intervention closed spontaneously, and the remainder had spontaneously decreased in size on follow-up. CONCLUSIONS: ASD is a relatively benign malformation in which early and noninvasive diagnosis can be achieved, with an extremely low interventional mortality. PMID- 10365360 TI - Alcohol use by parents who present their child to a paediatric emergency department. AB - OBJECTIVES: Parental alcohol misuse exposes children to risk, but is poorly identified in paediatric settings. We aimed to (i) measure the alcohol use in parents of a sample of children attending emergency and (ii) assess the quality of documentation of this alcohol use by paediatric residents. METHOD: Parents of children presenting to the Emergency Department of a children's hospital were interviewed regarding their alcohol use, and each completed an AUDIT CORE questionnaire. The child's case notes were reviewed for documentation of parental alcohol use. RESULTS: One hundred and ninety-three parents were interviewed. Although per capita alcohol use for both sexes, and hazardous drinking for females were found to be significantly less than general population figures, rates of hazardous drinking were high for fathers. Yet only 1% of case notes reviewed contained any documentation of parental alcohol use. CONCLUSION: This small study suggests that although average alcohol intake amongst parents presenting their children to a paediatric emergency department may be lower than for the general population, there is a significant prevalence of undetected hazardous drinking amongst parents. PMID- 10365361 TI - Confirmation that Child Behavior Checklist clinical scales discriminate juvenile mania from attention deficit hyperactivity disorder. AB - OBJECTIVE: To determine whether boys meeting diagnostic criteria for juvenile mania and attention deficit hyperactivity disorder (mania-ADHD) may be distinguished from boys with ADHD alone on a range of clinical and family variables. METHODOLOGY: Boys aged 9-13 years with mania-ADHD (n = 25), ADHD alone (n = 99), or no psychiatric diagnosis (n = 27) were compared on parent and teacher report Child Behavior Checklists (CBCL) and Conners Questionnaires, self report CBCLs, patterns of comorbidity, intellectual functioning, and family variables. RESULTS: Mania-ADHD subjects had significantly higher mean ratings than ADHD only subjects on the parent CBCL for the Withdrawn, Thought Problems, Delinquent Behavior and Aggressive Behavior scales and significantly higher rates of comorbid depression, anxiety and psychotic symptoms. Other variables did not distinguish the mania-ADHD and ADHD only groups. CONCLUSIONS: These data confirm previous research indicating that the CBCL may be used to assist in the clinical identification of manic children. PMID- 10365362 TI - Simultaneous presentation of coeliac disease and ulcerative colitis in a child. AB - Coeliac disease and inflammatory bowel disease (IBD) individually are not uncommon in children, but the occurrence of both conditions together is rare. The combined presentation of coeliac disease and IBD in a girl of 7 years is presented with a review of the related literature. The occurrence of coeliac disease with IBD should be considered at the time of diagnosis and at relapse, or where there is difficulty maintaining remission in established IBD. Screening with serum antibody tests may be helpful. PMID- 10365363 TI - The first case of H5N1 avian influenza infection in a human with complications of adult respiratory distress syndrome and Reye's syndrome. AB - Avian influenza virus was not known to cause systemic infection in humans before. We report a 3-year-old boy with good past health who developed pneumonia caused by H5N1 avian influenza A virus (A/Hong Kong/156/97). The virus was isolated from a tracheal aspirate. There were complications of Reye's syndrome, adult respiratory distress syndrome, and multiple organ system failure. He had a history of receiving aspirin. His adult respiratory distress syndrome did not respond to endotracheal surfactant replacement therapy. He died 6 days after admission. Clinicians should be alert to the importance of a new human influenza strain. PMID- 10365364 TI - Risk of gonadoblastoma in female patients with Y chromosome abnormalities and dysgenetic gonads. AB - We report two female patients with gonadal dysgenesis and sex chromosome mosaicism involving the Y chromosome. Conventional karyotyping was supplemented with fluorescent in situ hybridisation techniques in order to confirm the presence of Y chromosomes. One patient is a phenotypic female with karyotype 45,X/46,X,idic(Y)(q11.2). She underwent a laparoscopic gonadectomy at which streak ovaries without evidence of gonadoblastoma were removed. The second patient presented as a virilised female with karyotype 45,X/47,XYY. At laparoscopy, she was found to have mixed gonadal dysgenesis with a gonadoblastoma in situ. We recommend early gonadectomy in female children presenting with gonadal dysgenesis and the presence of a Y chromosome although once the gonadoblastoma locus on Y chromosome gene has been cloned it may be possible to identify those patients who have a low risk of developing gonadoblastoma. PMID- 10365365 TI - Surfactant protein B deficiency: clinical, histological and molecular evaluation. AB - Congenital alveolar proteinosis due to surfactant protein B deficiency is an inherited disease which results in severe respiratory failure in term infants soon after birth. The pathophysiologic basis of this disease is now known to be an inability to synthesise adequate quantities of normally functioning surfactant protein B. We report a male infant with fatal respiratory failure of neonatal onset, and histopathological features typical of those seen in congenital alveolar proteinosis. Molecular analysis of genomic DNA revealed two mutations, the 'common' 121ins2 mutation in exon 4, and a novel 2bp frameshift mutation in exon 5. We believe this is the first Australian case of surfactant protein B deficiency confirmed by molecular analysis. PMID- 10365366 TI - Calvarial tuberculosis. AB - We report a six-year-old boy who presented with swelling of the forehead, and had calvarial tuberculosis, a rare form of tuberculous osteitis. PMID- 10365367 TI - Taking a swing at boxing. PMID- 10365368 TI - Serum sickness in a paediatric emergency department: the role of cefaclor. PMID- 10365369 TI - Recurrent late onset group B streptococcal infection with parotitis. PMID- 10365370 TI - Dose dependent changes in 74As-arsenate metabolism of Flemish Giant rabbits. AB - The metabolic handling of 74As-arsenate (As(V)) was studied in rabbits injected intraperitoneally (i.p.) with increasing doses of As(V) (0.00 to 1.00 mg As(V)/kg/day) over a period of 10 days. Plasma, packed cells, urine from the bladder and several tissues were analyzed for their 74As content and presence of 74As-As(V) metabolites 4 h after administration of 74As-As(V). 74As showed strong increases with increasing As(V) dose in nails and bone whereas in fat, thyroid and kidneys it decreased. Also with increasing As(V) dose, arsenate was less efficiently methylated to dimethylarsenic acid (DMA) and became more bound to insoluble tissue constituents. As a result 74As-DMA levels in tissues were systematically lower in the groups of rabbits receiving the higher doses, be it with a wide variation from one type of tissue to the other. The behaviour of 74As monomethylarsonic acid (MMA) was different. The levels did not decrease significantly, occasionally even increased compared to the control group, indicating that especially the second methylation step is sensitive towards increasing doses of As(V). 74As-arsenite (As(III)), formed by in vivo reduction of As(V), reached maximal levels in the 0.25 mg As(V)/kg/day group as a result of the inhibited methylation. At doses > 0.25 mg As(V)/kg/day the amount of 74As As(V) increased especially in plasma, packed cells and the urine in the bladder, indicative for a less efficient reduction of As(V). PMID- 10365371 TI - The effect of changing living standards on iron status in pregnancy in Calabar urban. AB - Reduced meat intake is often associated with iron deficiency anaemia. Reduced meat intake that arose from the frugality associated with a prolonged period of national economic reorientation policy, known as the "structural adjustment programme" (SAP), may have placed iron-stress on pregnancy in particular. Iron status of pre-SAP and SAP pregnancies were established from measurement of the haematological values of subjects. Indices such as haemoglobin (Hb), packed cell volume, serum Fe were lower in pregnancy especially during SAP. However, SAP pregnancy serum ferritin did not respond significantly to iron depletion. Unlike PCV, Total Iron Binding Capacity (TIBC) and serum transferrin values showed a reverse behaviour. The level of these values, however, portends a toll exerted on body iron status in pregnancy associated with reduced haem iron intake. The changes in these values did not, however, lead to any over symptoms of iron deficiency probably because increased sea food (molluscs) consumption ameliorates the iron-toll from low absorbed non-haem iron. For lower income rural women in non-coastal areas, the absence of seafood in their diets exacerbates the low iron status in Pregnancy. PMID- 10365372 TI - Effects of chronic alternating cadmium exposure on the episodic secretion of prolactin in male rats. AB - Cadmium increases or decreases prolactin secretion depending on the dose and duration of the exposure to the metal. However, whether there are cadmium effects on the episodic prolactin secretion is less well known. This study was undertaken to address whether chronic alternating exposure to two different doses of cadmium affects the episodic pattern of prolactin and to what extent the effects of cadmium are age-dependent. Male rats were treated s.c. with cadmium chloride (0.5 or 1.0 mg/kg) from day 30 to 60, or from day 60 to 90 of age, with alteration of the doses every 4 days, starting with the smaller dose. Controls received vehicle every 4 days. The last dose of cadmium was given 48 h prior to the pulsatility study. Prolactin secretion in the 4 experimental groups studied was episodic and changed significantly after cadmium exposure. Cadmium administration from day 30 to 60 of life significantly decreased the mean half-life of prolactin. On the other hand, when administered from day 60 to 90 cadmium significantly decreased the mean as well as serum prolactin levels and the absolute amplitude of the prolactin pulses, their duration, the relative amplitude or the mean half-life of the hormone. The frequency of prolactin peaks was not changed by cadmium administration. The results indicate that low intermittent doses of cadmium chronically administered change the episodic secretion pattern of prolactin in rats. The effects of cadmium on prolactin secretion were age dependent. PMID- 10365373 TI - Accumulation of metals in the teeth of inhabitants of two towns in the south of Poland. AB - Metals and Ca, Na and K were examined in human teeth taken from inhabitants of two towns Sosnowiec and Katowice in the south of Poland (n = 170 samples) during 1993/1994. They are located in Upper Silesia, a heavily industrialised and urbanised region of Poland. The concentrations of Pb, Mn, Fe, Cd, Cu, Ni, Co, Zn and Cr were determined by AAS, and Na, Ca and K by flame photometer. Variations in the Pb/Fe, Pb/Mn, Pb/Ca and Cd/Ca ratios of elements in human teeth were examined. Influence of age, sex type of teeth on concentrations of elements for inhabitants from the two different towns (Katowice and Sosnowiec) and on levels of ratios of the elements were analysed using one-way ANOVA. Cluster Analysis was used to investigate relationships among metals. The smallest duster distances were obtained for: Ca-Fe, Mn-Cd and Na-Co, which indicated the strongest relation between elements in teeth for the investigated population living in a heavily industrialised and urbanised region. PMID- 10365374 TI - Distribution of lithium in different CNS areas and other tissues of adult male and female vizcacha (Lagostomus maximus maximus). AB - In a previous study (1) we demonstrated that lithium administration (1.0 mmol/kg b.wt., per day for 4 weeks) in intact vizcacha (Lagostomus maximus maximus) leads to significant histological alterations in the kidneys, ovarie and testicles, while these three tissues were not damaged in rats. Male vizcachas died within 4 days when administered LiCl 3 mmol/kg b.wt., while females were not affected. The lithium renal clearance presented no changes in either males or females. The 1.0 mmol/kg b.wt. dose was used in the experiments (2). In this study we examined the distribution of lithium in various tissues of male and female vizcacha (Lagostomus maximus maximus) administered LiCl by injection (1 mmol/kg b.wt.) for one day (Group I) and thirty days (Group II). Blood sample was obtained after 24 hours (Group I) and 30 days (Group II). The tissues investigated were: pituitary, hypothalamus, cerebral cortex, cerebellum, corpus callous, small and large intestine, kidney and suprarenal. The concentration of lithium in tissues and serum was determined by atomic absortion spectrometry (3,4). In Group I a significant lithium concentration increment (mumol/g of tissue) was observed in all the tissues of male vizcachas as compared to female vizcacha. A similar distribution was obtained in animals treated for 30 days. In the pituitary, however this difference between males and females was not significant. The male lithium serum levels were significantly higher than those of female animals. In conclusion, we suggest that the particular structure of the cell membrane (e.g., number and characteristic of sodium channels) of each tissue and/or the intracellular mechanisms of transport, elimination and metabolism might explain the unequal lithium distribution and the difference recovery from the damage produced. The results suggest that the vizcacha could be a useful model for the study of lithium toxicity. PMID- 10365376 TI - Validation of a questionnaire method for estimating extent of menstrual blood loss in young adult women. AB - The objective of this study was to validate two indirect methods for estimating the extent of menstrual blood loss against a reference method to determine which method would be most appropriate for use in a population of young adult women. Thirty-two women aged 18 to 29 years (mean +/- SD; 22.4 +/- 2.8) were recruited by poster in Dunedin (New Zealand). Data are presented for 29 women. A recall method and a record method for estimating extent of menstrual loss were validated against a weighed reference method. Spearman rank correlation coefficients between blood loss assessed by Weighed Menstrual Loss and Menstrual Record was rs = 0.47 (p = 0.012), and between Weighed Menstrual Loss and Menstrual Recall, was rs = 0.61 (p = 0.001). The Record method correctly classified 66% of participants into the same tertile, grossly misclassifying 14%. The Recall method correctly classified 59% of participants, grossly misclassifying 7%. Reference method menstrual loss calculated for surrogate categories demonstrated a significant difference between the second and third tertiles for the Record method, and between the first and third tertiles for the Recall method. The Menstrual Recall method can differentiate between low and high levels of menstrual blood loss in young adult women, is quick to complete and analyse, and has a low participant burden. PMID- 10365375 TI - Glutathione peroxidase activity modulates fatty acid profiles of plasma and breast milk in Chinese women. AB - Since little is known about the effect of selenium on the fatty acid profiles (FAP) of human breast milk, the purpose of this study was to measure the effect of habitual dietary selenium (Se) intake on this profile in plasma and breast milk. Subjects were lactating women from three locations in China where habitual selenium intakes are extremely low (Xichang), adequate (Beijing), or extremely high (Enshi). Plasma and milk samples were obtained within seven days of parturition (early samples) or within eighteen months postpartum (mature samples) and analyzed for selenium concentration, glutathione peroxidase (Gpx) activity and FAP. Plasma and milk selenium concentrations were significantly lower in the samples from women from Xichang and significantly higher in those from Enshi when compared to those from Beijing. Plasma Gpx activity, however, was higher in samples from Beijing than Xichang or Enshi. In contrast, the early breast milk samples had similar Gpx activity regardless of location. The mature samples, however, followed the same trend as plasma with the samples obtained from the women in Beijing having the highest activity. Of the unsaturated fatty acids examined, the concentration of linoleic acid, 18:2(n-6), in both plasma and milk was greater in the samples from Beijing when compared to those from Xichang or Enshi. Thus dietary selenium appears to influence the fatty acid composition in human breast milk, but influences Gpx activity only in mature milk samples. PMID- 10365378 TI - Direct determination of selenium and other trace elements in serum samples by ICP MS. AB - Selenium belongs to a group of trace elements of special interest in biological samples for clinical diagnosis. Selenium has antioxidizing functions and is essential for providing the organism with triiodothyronine produced from thyroxine. Among several analytical techniques used to determine the Se concentration in serum, Inductively Coupled Plasma Mass Spectrometry (ICP-MS) has been used in the past because of its high sensitivity. Interference problems originating from different ions on the major Se isotopes have been described to be a limiting factor for the direct determination of Se in these matrices. Standard addition calibration or isotope dilution is often required to overcome carbon-enhanced ionisation effects in biological sample matrices. In most cases, the typical serum sample volume which is available for the analysis is limited to 0.5 ml or less, making multiple sample preparation for standard addition calibration impractical. Isotope dilution requires enriched isotopes and substantial sample preparation. Furthermore, the approximate Se concentration in every sample has to be known to adjust the appropriate amount of spike to each sample. Matrix matching with methanol has been described to overcome ionisation effects but we found limiting factors of this application when other trace elements are also determined within one sample run. This paper describes an effective sample preparation method which allows the direct determination of Se in serum without limiting the analytical capabilities for the additional determination of Al, Cu, Ni, Co, Cd, Mn and Zn in a single sample run by ICP-MS. Optimization procedures are presented and results of the analysis of reference samples are discussed, with a comparison of more than 150 serum data with those obtained by the GF-AAS method. PMID- 10365377 TI - Na2Zn3(CO3)4.3H2O: a potentially useful compound for zinc supplementation. AB - The infrared and Raman spectra of the title compound were recorded and are briefly discussed on the basis of its structural characteristics. Its thermal behaviour was investigated by means of TG and DTA measurements. Several dissolution tests were also performed. The results support the potential usefulness of this double carbonate as a useful compound for Zn(II) supplementation. PMID- 10365379 TI - Review of publications. PMID- 10365380 TI - [Genetic polymorphisms of the renin-angiotensin system and essential hypertension]. AB - BACKGROUND: The renin-angiotensin system (RAS) plays an important role in blood pressure (BP) regulation. A number of RAS polymorphisms have been linked to essential hypertension (EH), but there is uncertainty about this association in other studies. We examined whether the insertion/deletion (I/D) polymorphism of the angiotensin converting enzyme (ACE) gene, and the M235T and T174M polymorphisms of the angiotensinogen (AGT) gene are associated with EH in a Spanish sample of hypertensive patients. MATERIAL AND METHODS: We studied 75 patients with EH (BP > 160/100 mmHg), aged 55 (8.5) years, 30 males, systolic BP (SBP) 182 +/- (22.1) mmHg, diastolic BP (DBP) 109 +/- (9.9) mmHg (mean [SD]) and a strong family history of the disease. As a control group, 75 healthy subjects with no family history of cardiovascular disease were studied. The polymorphisms were determined by PCR amplification of genomic DNA, followed by enzyme digestion for the AGT gene polymorphisms. RESULTS: The genotype distribution and the frequencies of the alleles of the three RAS polymorphisms were similar in hypertensive and control subjects. In addition, we did not find any compound effect of the I/D ACE gene and M235T AGT gene polymorphisms on BP levels in hypertensive and control subjects. CONCLUSIONS: In this sample, the contribution of the ACE I/D polymorphism and the AGT M235T and T174M polymorphisms in the development of EH seems to be less important than previously estimated. PMID- 10365381 TI - [Philadelphia chromosome-positive chronic myeloid leukemia in the elderly: presenting features, natural history and survival]. AB - BACKGROUND: Among patients with chronic myeloid leukaemia (CML) old people represent a minority whose disease characteristics are not well known. The aim of the study was to analyze the presenting features, the evolutive course, and the survival of older persons with Ph-positive CML. PATIENTS AND METHODS: Forty-four individuals > 65 years diagnosed with Ph-positive CML in a single centre were compared with 292 younger patients. RESULTS: Comparison of the presenting features of chronic phase Ph-positive CML patients yielded the following significant differences: predominance of female sex (15 males/29 females versus 155/137; p = 0.02), higher proportion of patients with anaemic syndrome (12% versus 2%; p = 0.001), lower frequency of splenomegaly (41% versus 68%; p = 0.001), and higher serum levels of uric acid (p = 0.0006) in the older group. Although the latter patients survived significantly less (median survival 36.6 months, 95% CI: 27-46.2, versus 57.6 months, 95%: 51.2-64.1; p = 0.004), 9 of the 33 deaths registered in this group (27%) occurred in the chronic phase of CML, versus 15 (9%) of the 166 deaths in the younger group (p = 0.003). When chronic phase deaths were excluded and leukaemia-related deaths only considered (i.e., those occurring in the BC or the accelerated phase of CML), old patients still had a shorter survival but the difference was no longer significant. CONCLUSIONS: Ph-positive CML features are essentially the same in older and young individuals, since most of the differences observed are attributable rather to the patients' advanced age than to the leukaemia itself. PMID- 10365383 TI - [How will diabetes be diagnosed after the year 2000 in Spain?]. PMID- 10365382 TI - [Dynamics of immunoglobulin production in non infected children born from HIV-1 infected mothers: effect of zidovudine]. AB - BACKGROUND: To evaluate the possible effect of zidovudine (ZDV) on inmunoglobins production in infants born to HIV-1 infected women. SUBJECTS AND METHODS: We have studied the immunoglobulins serum levels in 57 non-infected children born to HIV infected mothers. The children were divided into two groups: group A, 28 children born to HIV-1 infected mothers that received ZDV on protocol 076 conditions, and group B, 29 children born to mothers that did not receive anti-HIV-1 drugs. Quantification of serum IgG, IgA and IgM was performed by nephelometric techniques. RESULTS: The median time to reach normal IgA values at 12 months, was 25.57 months (confidence interval [CI] 95%: 22.01-29.12) in the children of group A and 12.67 months (CI 95%: 9.90-15.44) in the children of group B (p = 0.01). The median time to reach normal IgM values at 12 months was 15.93 months (CI 95%: 15.21-16.65) in group A children versus 11.20 months (CI 95%: 8.51-13.89) in group B (p = 0.11). The median time to reach normal IgG values at 12 months was 19.67 months (CI 95%: 13.12-16.22) in group A children versus 12.73 months (CI 95%: 11.16-14.30) in group B (p = 0.05). The normal IgA levels were reached 2.36 (CI 95%: 1.16-4.81) times later in group A than in group B children (p = 0.02), whereas normal IgG levels were reached 1.88 (CI 95%: 0.94-3.78) times later in group A than in group B of children. CONCLUSIONS: Our results indicate that treatment of pregnant mothers with ZDV clearly affect the ability of their newborns to produce inmunoglobulins, which may have important practical implications for their vaccination protocols. PMID- 10365384 TI - [Granulocyte transfusions. Return to the past]. PMID- 10365386 TI - [Productivity patterns in medical departments. Working Group of Medical Services of Community Hospitals, Catalonia]. PMID- 10365385 TI - [Non-gastric mucosa-associated lymphoid tissue (MALT) lymphomas: analysis of 14 patients]. AB - BACKGROUND: Mucosa-associated lymphoid tissue (MALT) lymphomas are a well defined group of B-cell non-Hodgkin's lymphomas, that arise in a wide variety of extranodal sites, most frequently in the stomach and related to Helicobacter pylori infection. The aim of the present study was to analyze the presenting features, natural history and outcome in 14 patients with non-gastric MALT lymphoma. PATIENTS AND METHODS: The main clinical data, treatment and outcome were recorded for the 14 patients with non-gastric MALT lymphoma diagnosed at a single institution in a 12 year period. The median age was 68 years and 13 patients were females. Diagnosis was made according to the REAL classification criteria. RESULTS: The initial location was thyroid (3 patients), parotid (three), submaxilar gland (three), skin (two), Waldeyer's ring (one), breast (one), lung (one), small bowel (one), liver (one) and ovary (one). At diagnosis 3 patients had > or = 2 extranodal involved sites. Autoimmune disorders were present in 5 patients: Hashimoto's thyroiditis (three), Sjogren's syndrome (one) and both (one). Two patients had a poor performance status (ECOG > 1) and B symptoms. Five patients (36%) were in stage IV, two of them because of bone marrow infiltration. All patients had a normal serum LDH level, and 5 had high beta 2-microglobulin level. The treatment consisted in surgical resection (2 patients), surgery and radiotherapy (one), surgery and chemotherapy (two), chemotherapy and radiotherapy (two) and chemotherapy alone (7 patients, three of them with doxorubicin-containing regimens). Twelve patients were evaluable for response. Complete response, partial response and failure rates were 75, 17 and 8%, respectively. Two of the 11 responders progressed, one of them with advanced stage disease. The actuarial 4-year disease-free survival was 77% (CI 95%: 47 100%). After a median follow-up of 3.4 years, 100% of the patients were alive. CONCLUSION: Non-gastric MALT lymphomas may be associated with autoimmune disorders, may present as disseminated disease and have a very good outcome. PMID- 10365387 TI - [The size of the effect of the difference between two means: statistically significant or clinically relevant?]. PMID- 10365388 TI - [Tobacco and cancer: from epidemiological association to molecular evidence]. PMID- 10365389 TI - [The safety of the derivatives of retinoic acid administered topically during pregnancy]. PMID- 10365390 TI - [Meningoencephalitis: an unusual complication of influenza vaccine]. PMID- 10365391 TI - [Sarcoidosis in a patient with systemic sclerosis]. PMID- 10365392 TI - [Hematopoietic growth factors and anaphylaxis]. PMID- 10365393 TI - [Anorectal Hodgkin disease in an HIV-infected patient]. PMID- 10365394 TI - [Bronchospasm caused by nebulized solution of salbutamol]. PMID- 10365395 TI - Genetic control and dynamics of the cellular immune response to the human T-cell leukaemia virus, HTLV-I. AB - About 1% of people infected with the human T-cell leukaemia virus, type 1 (HTLV I) develop a disabling chronic inflammatory disease of the central nervous system known as HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP). Patients with HAM/TSP have a vigorous immune response to HTLV-I, and it has been widely suggested that this immune response, particularly the HTLV-I-specific cytotoxic T-lymphocyte (CTL) response, causes the tissue damage that is seen in HAM/TSP. In this paper we summarize recent evidence that a strong CTL response to HTLV-I does in fact protect against HAM/TSP by reducing the proviral load of HTLV I. We conclude that HTLV-I is persistently replicating at a high level, despite the relative constancy of its genome sequence. These results imply that antiretroviral drugs could reduce the risk of HAM/TSP by reducing the viral load, and that an effective anti-HTLV-I vaccine should elicit a strong CTL response to the virus. The dynamic nature of the infection also has implications for the epidemiology and the evolution of HTLV-I. PMID- 10365396 TI - Bacterial population genetics, evolution and epidemiology. AB - Asexual bacterial populations inevitably consist of an assemblage of distinct clonal lineages. However, bacterial populations are not entirely asexual since recombinational exchanges occur, mobilizing small genome segments among lineages and species. The relative contribution of recombination, as opposed to de novo mutation, in the generation of new bacterial genotypes varies among bacterial populations and, as this contribution increases, the clonality of a given population decreases. In consequence, a spectrum of possible population structures exists, with few bacterial species occupying the extremes of highly clonal and completely non-clonal, most containing both clonal and non-clonal elements. The analysis of collections of bacterial isolates, which accurately represent the natural population, by nucleotide sequence determination of multiple housekeeping loci provides data that can be used both to investigate the population structure of bacterial pathogens and for the molecular characterization of bacterial isolates. Understanding the population structure of a given pathogen is important since it impacts on the questions that can be addressed by, and the methods and samples required for, effective molecular epidemiological studies. PMID- 10365397 TI - The effects of host heterogeneity on pathogen population structure. AB - We have shown that among pathogens, populations may self-organize into strains with non-overlapping repertoires of antigenic variants as a consequence of strong immune selection operating on polymorphic antigens. Recently, we have also demonstrated that over a wide range of intermediate levels of immune selection, pathogens may still be structured into discrete strains, but different sets of non-overlapping pathogen types will replace each other in a cyclical or chaotic manner. These models assume that the ranking of antigens in terms of the strength of the induced immune response is the same for every host. However, host immune responses may be restricted by the genotype of the individual. To explore this issue, a mathematical model was constructed under the assumption that a proportion of the host population responds principally to a variable antigen while the remainder of the population responds principally to a conserved antigen. The results of this analysis indicate that discrete strain structure (DSS) will be maintained even with a high frequency of hosts that do not respond in a variant-specific manner. Furthermore, the range of the immune selection pressure over which DSS prevails is increased (and the region of cyclical or chaotic behaviour reduced) by the inclusion of hosts that respond in a cross reactive rather than a variant-specific manner. PMID- 10365398 TI - Studies of antibiotic resistance within the patient, hospitals and the community using simple mathematical models. AB - The emergence of antibiotic resistance in a wide variety of important pathogens of humans presents a worldwide threat to public health. This paper describes recent work on the use of mathematical models of the emergence and spread of resistance bacteria, on scales ranging from within the patient, in hospitals and within communities of people. Model development starts within the treated patient, and pharmacokinetic and pharmacodynamic principles are melded within a framework that mirrors the interaction between bacterial population growth, drug treatment and the immunological responses targeted at the pathogen. The model helps identify areas in which more precise information is needed, particularly in the context of how drugs influence pathogen birth and death rates (pharmacodynamics). The next area addressed is the spread of multiply drug resistant bacteria in hospital settings. Models of the transmission dynamics of the pathogen provide a framework for assessing the relative merits of different forms of intervention, and provide criteria for control or eradication. The model is applied to the spread of vancomycin-resistant enterococci in an intensive care setting. This model framework is generalized to consider the spread of resistant organisms between hospitals. The model framework allows for heterogeneity in hospital size and highlights the importance of large hospitals in the maintenance of resistant organisms within a defined country. The spread of methicillin resistant Staphylococcus aureus (MRSA) in England and Wales provides a template for model construction and analysis. The final section addresses the emergence and spread of resistant organisms in communities of people and the dependence on the intensity of selection as measured by the volume or rate of drug use. Model output is fitted to data for Finland and Iceland and conclusions drawn concerning the key factors determining the rate of spread and decay once drug pressure is relaxed. PMID- 10365400 TI - Population dynamics of scrapie in a sheep flock. AB - A detailed analysis of an outbreak of natural scrapie in a flock of Cheviot sheep is described. A total of 137 cases was reported over 13 years among 1307 sheep born into the flock. The epidemiology of scrapie can only be understood with reference to sheep demography, the population genetics of susceptibility to scrapie, pathogenesis during a long incubation period, and the rate of transmission (by both vertical and horizontal routes), all of which interact in complex ways. A mathematical model incorporating these features is described, parameter values and model inputs are derived from available information from the flock and from independent sources, and model outputs are compared with the field data. The model is able to reproduce key features of the outbreak, including its long duration and the ages of cases. The analysis supports earlier work suggesting that many infected sheep do not survive to show clinical signs, that most cases arise through horizontal transmission, and that there is strong selection against susceptible genotypes. However, important aspects of scrapie epidemiology remain poorly understood, including the possible role of carrier genotypes and of an environmental reservoir of infectivity, and the mechanisms maintaining alleles giving susceptibility to scrapie in the sheep population. PMID- 10365399 TI - Antimalarial drug resistance and combination chemotherapy. AB - Antimarial drug resistance develops when spontaneously occurring parasite mutants with reduced susceptibility are selected, and are then transmitted. Drugs for which a single point mutation confers a marked reduction in susceptibility are particularly vulnerable. Low clearance and a shallow concentration-effect relationship increase the chance of selection. Use of combinations of antimalarials that do not share the same resistance mechanisms will reduce the chance of selection because the chance of a resistant mutant surviving is the product of the per parasite mutation rates for the individual drugs, multiplied by the number of parasites in an infection that are exposed to the drugs. Artemisinin derivatives are particularly effective combination partners because (i) they are very active antimalarials, producing up to 10,000-fold reductions in parasite biomass per asexual cycle; (ii) they reduce malaria transmissibility; and (iii) no resistance to these drugs has been reported yet. There are good arguments for no longer using antimalarial drugs alone in treatment, and instead always using a combination with artemisinin or one of its derivatives. PMID- 10365401 TI - Transmission dynamics and epidemiology of dengue: insights from age-stratified sero-prevalence surveys. AB - The relationship between infection with the four major serotypes of dengue virus and the occurrence of different forms of disease is complex and not fully understood. Interpreting longitudinal records of the incidence of serious disease to gain insight into the transmission dynamics and epidemiology of the virus is therefore complicated. Since age reflects duration of exposure, age-stratified, strain-specific serological surveys carried out at one point in time, or over a short time interval, can potentially provide a rich source of information on longitudinal patterns. This paper describes the development and application (to data collected in Thailand) of statistically rigorous methods designed to estimate time-varying, strain-specific forces of infection, and thus basic reproduction numbers, from cross-sectional serological data. The analyses provide support for the hypothesis that antibody-dependent enhancement of transmission influences observed epidemiological pattern. Age-stratified serological data also reveal evidence of a propensity for the annual incidence of infection to oscillate over time with a frequency of several years. The latter observation is consistent with the predictions of simple mathematical models of the transmission dynamics of the virus. The estimates of the basic reproduction numbers obtained are similar in magnitude for each dengue serotype, being in the range of four to six. Such values are higher than those obtained from earlier analyses, and the implications of this for dengue control are discussed. PMID- 10365402 TI - Stochastic dynamics and a power law for measles variability. AB - Since the discovery of a power law scaling between the mean and variance of natural populations, this phenomenon has been observed for a variety of species. Here, we show that the same form of power law scaling also occurs in measles case reports in England and Wales. Remarkably this power law holds over four orders of magnitude. We consider how the natural experiment of vaccination affects the slope of the power law. By examining simple generic models, we are able to predict the effects of stochasticity and coupling and we propose a new phenomenon associated with the critical community size. PMID- 10365403 TI - The epidemiology of pneumococcal infection in children in the developing world. AB - Pneumonia causes about three million deaths a year in young children, nearly all of which are in developing countries. Streptococcus pneumoniae (the pneumococcus) is the most important bacterial cause of pneumonia in young children and so is likely to be responsible for a high proportion of these deaths. The pneumococcus is also responsible for a substantial proportion of the 100,000-500,000 deaths that occur from meningitis in children each year. The incidence of invasive pneumococcal disease in children in the developing world is several times higher than in industrialized countries. This discrepancy may, in part, be due to socio economic differences but genetic factors may also play a role. Children with sickle cell disease have a substantially increased risk of invasive pneumococcal infection and a search is being made for other possible genetic risk factors. Infection with human immunodeficiency virus (HIV) also predisposes to invasive pneumococcal disease and so the incidence of this disease in young children is expected to rise as increasing numbers of African and Asian children are born with a perinatally acquired HIV infection. Until recently, pneumococcal infections could be treated effectively with penicillin, a cheap and safe antibiotic. However, pneumococci that are resistant to penicillin are becoming prevalent in many countries, necessitating a change to more costly antibiotics which may be beyond the reach of the health services of poor, developing countries. The spread of antibiotic resistance has provided an added stimulus to the development of vaccines that might be able to prevent pneumococcal disease in infants. Recently developed polysaccharide-protein conjugate vaccines show promise and are now undergoing field trials. How deployment of these vaccines will influence the balance between invasive pneumococcal infections and asymptomatic nasopharyngeal carriage of pneumococci is uncertain. PMID- 10365404 TI - The transmission dynamics of gonorrhoea: modelling the reported behaviour of infected patients from Newark, New Jersey. AB - A survey of the sexual behaviour of gonorrhoea patients in Newark was undertaken to evaluate parameters within a model of gonorrhoea transmission. Modelling work aimed to explain observed epidemiological patterns and to explore the potential impact of interventions. Reported behaviours, along with values derived from the literature, were used within a standard deterministic model of gonorrhoea transmission, where the population was stratified according to sex and rates of sex-partner change. The behaviours reported, particularly among women, are insufficient by themselves to explain the continued existence of gonorrhoea within the population. The majority of symptomatic patients seek treatment within a few days, and report that they do not have unprotected sex while symptomatic. The proportion of patients with low numbers of sex partners suggests that sexual mixing between people categorized according to sexual behaviour is near random. To explain the persistence of gonorrhoea, there must be some patients who, when infected, do not seek care in public clinics. In addition, gonorrhoea incidence in the model is sensitive to change, such that very small reductions in risk behaviour could lead to its elimination. This does not accord with the observed failure of many interventions to eliminate infection, suggesting that the modelled infection is too sensitive to change. The model, which has been influential in gonorrhoea epidemiology, is not consistent with the observed epidemiology of gonorrhoea in populations. Alternative models need to explore the observed stability of gonorrhoea before robust modelling conclusions can be drawn on how best to control infection. However, the current results do highlight the potential importance of asymptomatic infections and infections in those who are diseased and do not attend public health services. Screening and contact-tracing to identify asymptomatic infections in both men and women will be more effective in reaching those who maintain the infection within the community rather than simply treating symptomatic cases. PMID- 10365405 TI - Aggregation and distribution of strains in microparasites. AB - Recent research has shown that many parasite populations are made up of a number of epidemiologically distinct strains or genotypes. The implications of strain structure or genetic diversity for parasite population dynamics are still uncertain, partly because there is no coherent framework for the interpretation of field data. Here, we present an analysis of four published data sets for vector-borne microparasite infections where strains or genotypes have been distinguished: serotypes of African horse sickness (AHS) in zebra; types of Nannomonas trypanosomes in tsetse flies; parasite-induced erythrocyte surface antigen (PIESA) based isolates of Plasmodium falciparum malaria in humans, and the merozoite surface protein 2 gene (MSP-2) alleles of P. falciparum in humans and in anopheline mosquitoes. For each data set we consider the distribution of strains or types among hosts and any pairwise associations between strains or types. Where host age data are available we also compare age-prevalence relationships and estimates of the force of infection. Multiple infections of hosts are common and for most data sets infections have an aggregated distribution among hosts with a tendency towards positive associations between certain strains or types. These patterns could result from interactions (facilitation) between strains or types, or they could reflect patterns of contact between hosts and vectors. We use a mathematical model to illustrate the impact of host-vector contact patterns, finding that even if contact is random there may still be significant aggregation in parasite distributions. This effect is enhanced if there is non-random contact or other heterogeneities between hosts, vectors or parasites. In practice, different strains or types also have different forces of infection. We anticipate that aggregated distributions and positive associations between microparasite strains or types will be extremely common. PMID- 10365407 TI - Malaria epidemiology and economics: the effect of delayed immune acquisition on the cost-effectiveness of insecticide-treated bednets. AB - An understanding of the epidemiology of a disease is central in evaluating the health impact and cost-effectiveness of control interventions. The epidemiology of life-threatening malaria is receiving renewed interest, with concerns that the implementation of preventive measures such as insecticide-treated bednets (ITNs) while protecting young children might in fact increase the risks of mortality and morbidity in older ages by delaying the acquisition of functional immunity. This paper aims to illustrate how a combined approach of epidemiology and economics can be used to (i) explore the long-term impact of changes in epidemiological profiles, and (ii) identify those variables that are critical in determining whether an intervention will be an efficient use of resources. The key parameters for determining effectiveness are the protective efficacy of ITNs (reduction in all-cause mortality), the malaria attributable mortality and the increased malaria-specific mortality risk due to delays in the acquisition of functional immunity. In particular, the analysis demonstrates that delayed immune acquisition is not a problem per se, but that the critical issue is whether it occurs immediately following the implementation of an ITN programme or whether it builds up slowly over time. In the 'worst case' scenario where ITNs immediately increase malaria-specific mortality due to reduced immunity, the intervention might actually cost lives. In other words, it might be better to not use ITNs. On the other hand, if reduced immunity takes two years to develop, ITNs would still fall into the category of excellent value for money compared to other health interventions, saving a year of life (YLL) at a cost of between US$25-30. These types of calculations are important in identifying the parameters which field researchers should be seeking to measure to address the important question of the net impact of delaying the acquisition of immunity through preventive control measures. PMID- 10365409 TI - [Cervical artery dissections: several questions remain unresolved]. PMID- 10365408 TI - [Prevention of vascular complications after cerebral ischemia of arterial origin. European Stroke and Australian Stroke Prevention in Reversible Ischemia Trial (ESPRIT): moderated coagulation, aspirin-dipyridamole combination or aspirin alone?]. PMID- 10365406 TI - Population biology of human onchocerciasis. AB - Human onchocerciasis (river blindness) is the filarial infection caused by Onchocerca volvulus and transmitted among people through the bites of the Simulium vector. Some 86 million people around the world are at risk of acquiring the nematode, with 18 million people infected and 600,000 visually impaired, half of them partially or totally blind. 99% of cases occur in tropical Africa; scattered foci exist in Latin America. Until recently control programmes, in operation since 1975, have consisted of antivectorial measures. With the introduction of ivermectin in 1988, safe and effective chemotherapy is now available. With the original Onchocerciasis Control Programme of West Africa coming to an end, both the new African Programme for Onchocerciasis Control and the Onchocerciasis Elimination Programme for the Americas, rely heavily on ivermectin self-sustained mass delivery. In consequence, the need for understanding the processes regulating parasite abundance in human and simuliid populations is of utmost importance. We present a simple mathematical framework built around recent analyses of exposure- and density-dependent processes operating, respectively, within the human and vector hosts. An expression for the basic reproductive ratio, R0, is derived and related to the minimum vector density required for parasite persistence in localities of West Africa in general and northern Cameroon in particular. Model outputs suggest that constraints acting against parasite establishment in both humans and vectors are necessary to reproduce field observations, but those in humans may not fully protect against reinfection. Analyses of host age-profiles of infection prevalence, intensity, and aggregation for increasing levels of endemicity and intensity of transmission in the Vina valley of northern Cameroon are in agreement with these results and discussed in light of novel work on onchocerciasis immunology. PMID- 10365410 TI - [Chronic obliterative arterial disease of the lower limbs in the coronary patient: prevalence and prognostic incidence. The Monica Toulouse register]. AB - PURPOSE: This study was aimed at evaluating the prevalence of peripheral arterial disease of the lower extremities and its prognostic value in a population of patients from the Haute-Garonne department, who were hospitalized for acute coronary artery disease. METHODS: Between 1985 and 1991, four thousands three hundred and sixty-eight patients (3,680 males and 688 females) presenting with acute coronary artery disease were included in the study. RESULTS: The prevalence of peripheral arterial disease of the lower extremities was 13.4%, increasing with age and being higher in male patients. In regard to patients hospitalized for acute myocardial infarction (n = 2,417), independent relationships were observed between the 28-day mortality and the following: patient's age (odds ratio: 1.02; 95% confidence interval: 1.01-1.04; P < 0.0005), female gender (odds ratio: 1.32; 95% confidence interval: 1.17-1.54; P < 0.002), inclusion in the study (odds ratio 0.95; 95% confidence interval: 0.90-0.99; P < 0.02), previous coronary artery disease (odds ratio: 2.88; 95% confidence interval: 2.32-3.48; P < 0.0001), and peripheral arterial disease (odds ratio: 1.61; 95% confidence interval: 1.26-2.06; P < 0.0001). CONCLUSION: The prevalence of peripheral arterial disease of the lower extremities is high in patients with acute coronary artery disease in both genders, whatever the age. This disease is therefore an independent marker of mortality for acute myocardial infarction. Easy diagnosis of peripheral arterial disease of the lower limbs by measurement of the ankle pressure index allows identification of patients prone to death from acute myocardial infarction. PMID- 10365411 TI - [Etiology of oligoarthritis in equatorial Africa. A retrospective study of 80 cases in Brazzaville, Congo]. AB - PURPOSE: The objective was to determine causes of oligoarthritis in patients hospitalized in a Central Africa teaching hospital. METHODS: We collected case reports of patients presenting with arthritis located in two or three joints. Analysis included the disease frequency, age, gender, and etiology. Etiology was defined according to common criteria used for the diagnosis of rheumatic diseases. RESULTS: Eighty patients (60 males and 20 females; age range: 7-80 years; mean age: 35.6 years) were included in the study. Altogether they represented 5.8% of in-patients and 14.3% of the patients suffering from arthritis. The following causes were observed: HIV-related arthritis, 21 cases (12 males and nine females; mean age: 32.2 years). Arthritis was asymmetrical, acute and non-erosive. Recovery was observed after administration of a non steroidal anti-inflammatory drug for 4-6-weeks. Gout: 17 patients (15 males and two females; mean age: 51.1 years). Gout was related with one or more of the following: alcoholism (16 cases), hypertension (nine cases), and obesity (three cases), and was chronic in 12 patients. Infection-related arthritis: 17 cases (five males and 12 females; mean age 25.3 years). Portal of entry was either cutaneous (five cases) or obstetrical (seven cases). Most of the time, the causative microorganism was Staphylococcus. Rheumatic fever: five patients (four males and one female; mean age 18 years). Juvenile chronic arthritis: three cases (one male and two females; mean age 12.6 years). Reactive arthritis: three cases (three males; mean age: 32). Etiology is still unknown for 14 cases. CONCLUSION: Oligoarthritis appears to be secondary to HIV or bacterial infection in young adults, while gout affects more often the elderly. This study confirms that spondylarthropathy is unusual in Sub-Saharan Africa. Furthermore, it emphasizes the existence of numerous cases in whom etiology of the diseases is unknown. PMID- 10365412 TI - [Cervical artery dissection: recent data physiopathologic hypotheses]. AB - INTRODUCTION: Cervicocranial arterial dissection is a major cause of cerebral infarction in young subjects. Traumatic and infectious factors are frequently suspected to be at the origin of cervicocranial artery dissection. However, they are usually too minor or too common to explain the vessel wall split-off. Underlying arteriopathy predisposing to dissections is therefore often suspected. CURRENT KNOWLEDGE AND KEY POINTS: The hypothesis of an underlying arteriopathy is based in certain cases on either the discovery of hereditary connective tissue disorders (or secondary signs of these diseases) or their frequent association with vascular and cardiac morphological abnormalities, thus suggesting extracellular matrix abnormalities. Current histological and biochemical data do not suggest the existence of a unique form of the disease but rather indicate the presence of various matrix abnormalities that could involve one of the fibrillar components or its enzymatic regulation. FUTURE PROSPECTS AND PROJECTS: Classification of dissections according to the various vascular wall alterations would therefore permit to better define recurrence and familial risks and to improve overall management of the patients. PMID- 10365413 TI - [Digestive system carcinoid tumors: treatment]. AB - INTRODUCTION: Non-metastatic digestive carcinoid tumors are treated surgically, allowing in most cases recovery. In patients with metastasis and intestinal primary tumor, resection of the latter is proposed to avoid occlusion. CURRENT KNOWLEDGE AND KEY POINTS: Cytoreductive surgery of liver metastasis should be both contemplated and discussed if 80 to 90% of the tumor can be resected. In all other patients, intravenous chemotherapy or hepatic arterial chemoembolization should be discussed if metastases are located mainly in the liver. Response rates related to both treatments reach 30% and 50-80%, respectively, without clearly proven benefit in regard to survival. However, carcinoid tumors are often slowly progressive and symptomatic treatment of the carcinoid syndrome is a major concern. Long-acting somatostatin analogs, particularly slow-release formulations, have greatly improved patients' management. Diarrhea and flushing are controlled by long-term treatment involving either octreotide or lanreotide, a recently available somatostatin analog with slow release, without major side effects even at high dosages. PERSPECTIVES: Randomized studies currently in progress are aimed at comparing these various therapeutic modalities. PMID- 10365414 TI - [Spontaneous dissecting aneurysm of the internal carotid artery]. AB - INTRODUCTION: Dissecting aneurysms of the internal carotid artery are due to arterial wall dissection caused by hematoma. We report a case of spontaneous dissection. EXEGESIS: A 65-year-old man presented with painful Horner's syndrome and hypoglossal palsy, without a history of arterial traumatism. Magnetic resonance imaging showed carotid artery dissection. CONCLUSION: Distal and subadventicial dissection can induce compression of adjacent nerves without modifications of the arterial lumen. This type of wall hematoma may not be detected by ultrasonography and angiography. Magnetic resonance imaging proves to be the best method of investigation and should be primarily advocated. Anticoagulation treatment is necessary. PMID- 10365415 TI - [Hyperreactive malarial splenomegaly in a European returning from Africa]. AB - INTRODUCTION: Hyper-reactive malarial splenomegaly (HMS) syndrome related to abnormal immunologic response to repeated malarial infections is unusual in European expatriates. EXEGESIS: We report the case of a 72-year-old white male patient who had been residing in the Congo and developed a typical clinical features of hyperactive malarial syndrome characterized by massive splenomegaly with hypersplenism, high titers of malarial IgM antibodies, IgM macroglobulinemia, liver and medullary lymphocytic proliferation, and a clinical and immunological response to long-term chloroquine therapy. CONCLUSION: Criteria for the diagnosis of hyper-reactive malarial splenomegaly are useful. However, making a distinction from malignant lymphoproliferative disorders is difficult, as a sustained response to chloroquine is required. Therefore, chloroquine appears to have a regulatory effect on the immune system. PMID- 10365416 TI - [Pseudallescheria boydii osteoarthritis in a patient with acute lymphoblastic leukemia: a case report]. AB - INTRODUCTION: The outcome of neutropenic patients with Pseudallescheria boydii infection is poor. EXEGESIS: We report the first case of Pseudallescheria boydii hip arthritis in a patient treated for acute lymphoblastic leukemia. In vitro susceptibility testing showed that the strain was resistant to amphotericin B, fluorocytosine and nystatin, but susceptible to itraconazole. The patient received oral itraconazole (600 mg/day) and clinical symptoms initially resolved. Two months later, after a course of chemotherapy and high-dose steroids while receiving oral itraconazole treatment, the patient developed fever, skin lesions and disseminated lung infiltrates due to Pseudallescheria boydii and finally died. CONCLUSION: This case illustrates the severity of fungal infections due to Pseudallescheria boydii despite a presumably well-conducted antifungal therapy. PMID- 10365417 TI - [A not very Catholic nodule?]. PMID- 10365418 TI - [Current knowledge of the physiology and physiopathology of dendritic cells]. PMID- 10365419 TI - [Unmeasurable uric acid in blood and urine; xanthine dehydrogenase deficiency (or hereditary xanthinuria)]. PMID- 10365420 TI - [Multiple arterial thromboses from an intracardial embolism due to a malposition of a pacing lead]. PMID- 10365421 TI - [Localized Fusarium infection in acute myeloid leukemia]. PMID- 10365422 TI - [Thrombotic microangiopathy associated with cancer is an oncologic emergency: a case report]. PMID- 10365423 TI - A multisensory integration model of human stance control. AB - A model is presented to study and quantify the contribution of all available sensory information to human standing based on optimal estimation theory. In the model, delayed sensory information is integrated in such a way that a best estimate of body orientation is obtained. The model approach agrees with the present theory of the goal of human balance control. The model is not based on purely inverted pendulum body dynamics, but rather on a three-link segment model of a standing human on a movable support base. In addition, the model is non linear and explicitly addresses the problem of multisensory integration and neural time delays. A predictive element is included in the controller to compensate for time delays, necessary to maintain erect body orientation. Model results of sensory perturbations on total body sway closely resemble experimental results. Despite internal and external perturbations, the controller is able to stabilise the model of an inherently unstable standing human with neural time delays of 100 ms. It is concluded, that the model is capable of studying and quantifying multisensory integration in human stance control. We aim to apply the model in (1) the design and development of prostheses and orthoses and (2) the diagnosis of neurological balance disorders. PMID- 10365424 TI - A joint interspike interval difference stochastic spike train analysis: detecting local trends in the temporal firing patterns of single neurons. AB - We introduce a stochastic spike train analysis method called joint interspike interval difference (JISID) analysis. By design, this method detects changes in firing interspike intervals (ISIs), called local trends, within a 4-spike pattern in a spike train. This analysis classifies 4-spike patterns that have similar incremental changes. It characterizes the higher-order serial dependence in spike firing relative to changes in the firing history. Mathematically, this spike train analysis describes the statistical joint distribution of consecutive changes in ISIs, from which the serial dependence of the changes in higher-order intervals can be determined. It is similar to the joint interspike interval (JISI) analysis, except that the joint distribution of consecutive ISI differences (ISIDs) is quantified. The graphical location of points in the JISID scatter plot reveals the local trends in firing (i.e., monotonically increasing, monotonically decreasing, or transitional firing). The trajectory of these points in the serial-JISID plot traces the time evolution of these trends represented by a 5-spike pattern, while points in the JISID scatter plot represent trends of a 4 spike pattern. We provide complete theoretical interpretations of the JISID analysis. We also demonstrate that this method indeed identifies firing trends in both simulated spike trains and spike trains recorded from cultured neurons. PMID- 10365425 TI - Multiple-input, multiple-output system identification for characterization of limb stiffness dynamics. AB - This study presents time-domain and frequency-domain, multiple-input, multiple output (MIMO) linear system identification techniques that can be used to estimate the dynamic endpoint stiffness of a multijoint limb. The stiffness of a joint or limb arises from a number of physiological mechanisms and is thought to play a fundamental role in the control of posture and movement. Estimates of endpoint stiffness can therefore be used to characterize its modulation during physiological tasks and may provide insight into how the nervous system normally controls motor behavior. Previous MIMO stiffness estimates have focused upon the static stiffness components only or assumed simple parametric models with elastic, viscous, and inertial components. The method presented here captures the full stiffness dynamics during a relatively short experimental trial while assuming only that the system is linear for small perturbations. Simulation studies were performed to investigate the performance of this approach under typical experimental conditions. It was found that a linear MIMO description of endpoint stiffness dynamics was sufficient to describe the displacement responses to small stochastic force perturbations. Distortion of these linear estimates by nonlinear centripetal and Coriolis forces was virtually undetectable for these perturbations. The system identification techniques were also found to be robust in the presence of significant output measurement noise and input coupling. These results indicate that the approach described here will allow the estimation of endpoint stiffness dynamics in an experimentally efficient manner with minimal assumptions about the specific form of these properties. PMID- 10365426 TI - A model for steady isometric muscle activation. AB - The present model of the motoneuronal (MN) pool-muscle complex (MNPMC) is deterministic and designed for steady isometric muscle activation. Time-dependent quantities are treated as time-averages. The character of the model is continuous in the sense that the motor unit (MU) population is described by a continuous density function. In contrast to most already published models, the wiring (synaptic weight) between the input fibers to the MNPMC and the MNs (about which no detailed data are known) is deduced, whereas the input-force relation is given. As suggested by experimental data, this relation is assumed to be linear during MU recruitment, but the model allows other, nonlinear relations. The input to the MN pool is defined as the number of action potentials per second in all input fibers, and the excitatory postsynaptic potential (EPSP) conductance in MNs evoked by the input is assumed to be proportional to the input. A single compartment model with a homogeneous membrane is used for a MN. The MNs start firing after passing a constant voltage threshold. The synaptic current-frequency relation is described by a linear function and the frequency-force transformation of a MU by an exponential function. The sum of the MU contraction forces is the muscle force, and the activation of the MUs obeys the size principle. The model parameters were determined a priori, i.e., the model was not used for their estimation. The analysis of the model reveals special features of the activation curve which we define as the relation between the input normalized by the threshold input of the MN pool and the force normalized by the maximal muscle force. This curve for any muscle turned out to be completely determined by the activation factor, the slope of the linear part of the activation curve (during MU recruitment). This factor determines quantitatively the relation between MU recruitment and rate modulation. This property of the model (the only known model with this property) allows a quantification of the recruitment gain (Kernell and Hultborn 1990). The interest of the activation factor is illustrated using two human muscles, namely the first dorsal interosseus muscle, a small muscle with a relatively small force at the end of recruitment, and the medial gastrocnemius muscle, a strong muscle with a relatively large force at the end of recruitment. It is concluded that the present model allows us to reproduce the main features of muscle activation in the steady state. Its analytical character facilitates a deeper understanding of these features. PMID- 10365427 TI - Muscle activity in rapid multi-degree-of-freedom elbow movements: solutions from a musculoskeletal model. AB - The activity of certain muscles that cross the elbow joint complex (EJC) are affected by forearm position and forearm movement during elbow flexion/extension. To investigate whether these changes are based on the musculoskeletal geometry of the joint, a three-dimensional musculotendinoskeletal computer model of the EJC was used to estimate individual muscle activity in multi-degree-of-freedom (df) rapid (ballistic) elbow movements. It is hypothesized that this model could reproduce the major features of elbow muscle activity during multi-df elbow movements using dynamic optimal control theory, given a minimum-time performance criterion. Results from the model are presented and verified with experimental kinematic and electromyographic data from movements that involved both one-df elbow flexion/extension and two-df flexion/extension with forearm pronation/supination. The model demonstrated how the activity of particular muscles is affected by both forearm position and movement, as measured in these experiments and as previously reported by others. These changes were most evident in the flexor muscles and least evident in the extensor muscles. The model also indicated that, for specific one- and two-df movements, activating a muscle that is antagonistic or noncontributory to the movement could reduce the movement time. The major features of muscle activity in multi-df elbow movements appear to be highly dependent on the joint's musculoskeletal geometry and are not strictly based on neural influences or neuroanatomical substrates. PMID- 10365428 TI - A mathematical model of the adaptive control of human arm motions. AB - This paper discusses similarities between models of adaptive motor control suggested by recent experiments with human and animal subjects, and the structure of a new control law derived mathematically from nonlinear stability theory. In both models, the control actions required to track a specified trajectory are adaptively assembled from a large collection of simple computational elements. By adaptively recombining these elements, the controllers develop complex internal models which are used to compensate for the effects of externally imposed forces or changes in the physical properties of the system. On a motor learning task involving planar, multi-joint arm motions, the simulated performance of the mathematical model is shown to be qualitatively similar to observed human performance, suggesting that the model captures some of the interesting features of the dynamics of low-level motor adaptation. PMID- 10365429 TI - Biodegradation of diesel oil by cold-adapted microorganisms in presence of sodium dodecyl sulfate. AB - The effect of different concentrations of the anionic surfactant sodium dodecyl sulfate (SDS) on biodegradation of diesel oil was assessed during 32 days at 10 degrees C, under simulated environmental conditions, in liquid culture and in an alpine soil. Low SDS concentrations (50-100 mg l-1) significantly enhanced oil biodegradation by a psychrotrophic inoculum in liquid culture, whereas higher SDS concentrations (500-1000 mg l-1) inhibited hydrocarbon biodegradation. Oil biodegradation by the indigenous microorganisms in soil was inhibited at all SDS concentrations tested. The surfactant itself was rapidly biodegraded both in liquid culture and in soil. PMID- 10365430 TI - Effect of the alkyl chain length on the anaerobic biodegradability and toxicity of quaternary ammonium based surfactants. AB - The anaerobic biodegradability and toxicity to methanogenic gas production of different alkyl chain length homologs of quaternary ammonium based surfactants were examinated. Two series of these cationic surfactants were selected: alkyl trimethyl ammonium and alkyl benzyl dimethyl ammonium compounds. A simple anaerobic gas production test containing municipal digester solids as a source of anaerobic bacteria was used. Under the applied methanogenic conditions, the cationic surfactants tested showed a very poor primary biodegradation and no evidence of any extent of ultimate biodegradation was observed. The toxicity of quaternary ammonium based surfactants to methanogenic gas production decreased with increasing the alkyl chain length. PMID- 10365432 TI - Polychlorinated dibenzo-p-dioxins and related compounds: the blood levels of young Japanese women. AB - We investigated PCDDs and related compounds in the blood of young Japanese women, approximately 20 years of age, who had not yet had children, and discussed how the TEQ level of PCDDs and related compounds in their blood may affect the next generation. Means of total TEQ levels were 0.063 pg/g for whole blood basis and 21 pg/g for lipid basis. TEQ of PCDDs, PCDFs and coplanar PCBs accounted for about 43, 34 and 23% of the total TEQ in the whole blood basis, respectively. In the lipid basis, their values were about 44, 34 and 22%, respectively. Previously, we investigated PCDDs and related compounds levels in mother's breast milk, lymphocyte subpopulation and thyroid function of their children, and found negative correlations between the TEQ level of PCDDs and related compounds and CD4+/CD8+, and/or the TEQ level of PCDDs and related compounds and the T4 level in 36 mothers and children. Of these cases, the average age was approximately 28 years. PCDDs and related compounds may be related to immunopathy, such as atopic dermatitis. The effects of PCDDs and related compounds on babies of young Japanese women are important and must be further evaluated. PMID- 10365433 TI - PCDDs and PCDFs in food samples from Catalonia, Spain. An assessment of dietary intake. AB - Food samples from local markets and supermarkets of Tarragona (Catalonia, Spain) were analyzed for PCDD/F concentrations. On lipid basis, PCDD/F levels in meat, fish, eggs, and fats and oils were similar or lower than those found in other countries. By contrast, in general terms PCDD/F levels in milk, vegetables, and cereals showed higher levels than those previously reported. The total dietary PCDD/F intake by the population of Tarragona was estimated to be 210 pg I TEQ/day. This value is higher than the dietary intake of PCDD/Fs found in a number of surveys from other countries. However, when total dietary intake of PCDD/Fs was calculated including only fish and seafood, meat, milk and dairy products, eggs, and fats and oils, a dietary intake of PCDD/Fs of 117 pg I TEQ/day was obtained. This intake is in the same range than that reported for different regions and countries. The results of the present study show that food groups such as vegetables, fruits, and cereals should not be excluded to estimate the total dietary intake of PCDD/Fs by general populations, especially in those countries and/or regions in which their consumptions are notable. PMID- 10365434 TI - Induction of micronuclei in human lymphocytes after occupational exposure to ultrasound. AB - Micronucleus assay combined with Giemsa and DAPI staining was performed on blood samples of subjects occupationally exposed to ultrasound. Lymphocytes were cultivated in vitro for 72 h. At 44h cytochalasin-B was added in cultures. Frequencies of micronuclei in exposed subjects statistically significant increased compared to control. The frequency of micronucleated cells and micronuclei in exposed subjects shows interindividual variability. Using DAPI staining we observed signal-positive and signal-negative micronuclei. Percentage of signal-positive micronuclei varies between 0 and 66.7% and signal-negative micronuclei between 33.3% and 100%. This study indicate harmful effects of ultrasound on human genome, but further investigations are necessary. PMID- 10365435 TI - Transformation of a non-oestrogenic steroid metabolite to an oestrogenically active substance by minimal bacterial activity. AB - The majority of oestrogenic material excreted from humans and wildlife, and therefore released into sewers, is in a conjugated form. However, the finding of "free" oestrogens in sewage effluent suggests that these metabolites are somehow converted back into an active form, before or during passage through a sewage treatment process. When male fathead minnows (Pimephales promelas) were continuously exposed to oestradiol-3-glucuronide, in a continuous-flow system, it demonstrated no inherent oestrogenic activity. However, when fish were exposed to effluent generated from laboratory simulations of sewage treatment processes, to which had been added oestradiol-3-glucuronide, oestrogenic activity was observed, suggesting microbial activity was capable of degrading the steroid metabolite into a more potent oestrogen. Oestrogenic potency was determined by measuring changes in plasma vitellogenin (egg yolk precursor) concentrations and gonadosomatic index. The results suggest that inactive metabolites of steroids are very readily biotransformed into biologically active oestrogens. PMID- 10365436 TI - Environmental risk assessment for trisodium [S,S]-ethylene diamine disuccinate, a biodegradable chelator used in detergent applications. AB - Environmental safety data are presented for [S,S]-Ethylene Diamine Disuccinate ([S,S]-EDDS), a new, biodegradable, strong transition metal chelator. An environmental risk assessment for its use in detergent applications, which takes into account the chelating properties of [S,S]-EDDS, is proposed. A property of [S,S]-EDDS that distinguishes it from other strong transition metal chelators is its, "ready" and transparent (no recalcitrant metabolites) biodegradation profile. Because its sorption to activated sludge solids is low (Kp of 40 l/kg), removal of [S,S]-EDDS during sewage treatment, which is greater than 96% as determined by the Continuous Activated Sludge test, is mainly ascribed to biodegradation. At projected use volumes in detergent applications [S,S]-EDDS predicted steady-state concentration in rivers leaving the mixing zone will be below 5 micrograms/l due to rapid biodegradation. [S,S]-EDDS exhibits low toxicity to fish and Daphnia (both EC50S > 1000 mg/l). By contrast, due to limitation of the algal test for chelators apparent toxicity was observed (EC50 = 0.290 mg/l, NOEC--No observable Effect Concentration = 0.125 mg/l). Schowanek et al. [1] demonstrated that this is not toxicity sensu stricto but a chelation effect of trace metals in the test medium and of resulting essential nutrients limitation. This requires specific attention when the results of algal toxicity are to be extrapolated to a field situation to perform realistic risk assessment. Metal speciation calculations, using MINEQL+, show that at the predicted environmental concentrations of [S,S]-EDDS (1-5 micrograms/l), such a chelation effect would be insignificant. These calculations allow to estimate the NOEC for chelation effects in the field to be in the range of 0.250-0.500 mg/l, depending on the background water chemistry. These values are well above the laboratory NOEC. An environmental risk assessment was performed using the EUSES (1.0) program. EUSES is currently the EU recommended tool for conducting risk assessments (TGD 1995). It was applied to estimate the river water and soil concentrations from production, formulation and private use life stages. The estimated PEC/PNEC ratio in all relevant environmental compartments is smaller than 1, indicating "no immediate concern" at the anticipated usage level. PMID- 10365437 TI - Enhancement of PAH biodegradation in soil by physicochemical pretreatment. AB - The effects were studied of short-term heating of contaminated soil and its soaking in an organic solvent on the subsequent biodegradation of PAHs. In a clayey dredged sludge with a high organic-matter content (12%), heating at 120 degrees C for one hour increased the degree of degradation after 21 days of an aged PAH contamination from 9.5 +/- 0.7% to 27 +/- 5%. Lower temperatures resulted in smaller increases. The observed increase in biodegradation is caused by either transfer of PAHs from sorption sites with low desorption rates to those with high ones or transformation of slow-sorption sites into fast-sorption ones. Soaking of the above sludge in a 4:1 (v/v) acetone-water mixture increased the degree of degradation from 9.5 +/- 0.7% to 20.4 +/- 1.4%, probably as a result of dissolution of the PAHs in the pore liquid during soaking. Thermal pretreatment of a contaminated sandy soil with a low organic-matter content showed no significant effect on the degradation of aged PAHs. Soaking of the sandy soil increased the degradation of only PAHs of high molecular weight, namely from 24 +/- 5% to 48 +/- 7%. PMID- 10365438 TI - mdm-2 gene amplification in 3T3-L1 preadipocytes. AB - In this study the regulation of the murine double minute-2 (mdm-2) gene was examined in NIH 3T3-L1 preadipocytes. The 3T3-L1 cell line, under proper conditions, has the capacity to differentiate from fibroblasts into adipocytes [15]. A recent report demonstrated that mdm-2 overexpression could block myogenesis [12]. While examining the regulation of the mdm-2 gene during adipogenesis, it was discovered that 3T3-L1 cells possess a 36-fold elevation of mdm-2 mRNA relative to A31 cells, another immortalized Balb/c 3T3 fibroblast cell line that lacks the capacity to differentiate. Based on Southern blot analysis, the increase in mdm-2 mRNA was the result of a mdm-2 gene amplification. The level of Mdm-2 protein in undifferentiated 3T3-L1 cells was elevated relative to A31 fibroblasts and resulted from translation of mRNA transcripts initiating from the p53-independent P1 promoter. We also examined how mdm-2 and p53 levels changed as undifferentiated fibroblasts converted to adipocytes. While mdm-2 mRNA levels remained elevated, p53 mRNA, protein, and DNA-binding activity decreased. These results suggest that adipogenesis is unaffected by elevated Mdm-2 levels and that the overexpression of mdm-2 mRNA is predominantly p53 independent. PMID- 10365439 TI - Mouse C2 myoblast cells resist HVJ (Sendai virus)-mediated cell fusion in the proliferating stage but become capable of fusion after differentiation. AB - To investigate the mechanism of myoblast fusion, we attempted to prepare artificial myotubes of mouse C2 myoblast cells using the hemagglutinating virus of Japan (HVJ, Sendai virus). Proliferating C2 cells showed strong resistance to HVJ-mediated cell fusion and remained morphologically unchanged even though massive numbers of virions adsorbed onto their surface. They showed no membrane disruption, which occurs in the early stage of cell fusion induced by HVJ. These observations suggest that proliferating C2 cells are resistant to HVJ-mediated cell fusion. However, upon induction of differentiation, C2 cells gradually became capable of fusion induced by HVJ and then even generated heterokaryons with Ehrlich ascites tumor cells. When differentiated C2 cells that had become fusion-sensitive were treated with HVJ in the presence of EDTA, they did not fuse but degenerated, suggesting that their cell membranes were transiently disrupted by interaction with HVJ. These results suggest that the cell membranes of myoblasts change to a fusion-capable state during the process of differentiation. PMID- 10365440 TI - Characterization of loricrin regulation in vitro and in transgenic mice. AB - We have previously shown that the promoter of a 6.5 kb mouse loricrin clone contains a functional AP-1 element and directs tissue-specific, but not differentiation-specific, expression. We now report the isolation of a 14-kb genomic clone containing an additional 7 kb of genomic sequence. The additional sequences limit expression of a reporter construct to differentiated keratinocytes in culture. The expression of the 6.5-kb and 14-kb loricrin constructs were also analyzed in transgenic mice. Significantly, loricrin was found in all layers of the epidermis of the 6.5-kb transgenics, including basal and spinous cells. The expression of the 14-kb clone was indistinguishable from that of the endogenous gene, confirming that the additional sequences contain negative regulatory elements that restrict loricrin expression to the granular layer in vivo. In addition, we show the AP-1 element localized in the loricrin proximal promoter is necessary but not sufficient for expression of the loricrin gene in vivo in transgenic mice. Finally, to gain further insight into how AP-1 family members regulate expression of the loricrin gene, we co-transfected the loricrin reporter constructs with expression plasmids for various fos and jun family members and demonstrated that c-Fos/Jun-B heterodimers could mimic the differentiation-specific induction of loricrin. PMID- 10365441 TI - Differential mRNA display analysis of two related but functionally distinct rat insulinoma (RIN) cell lines: identification of CD24 and its expression in the developing pancreas. AB - To identify genes that might play a role in growth and differentiation of pancreatic beta-cells, we have applied the technique of differential mRNA display to the lineage-related, but functionally distinct rat insulinoma (RIN) cell lines RIN-5AH and RIN-A12. Direct comparison of PCR-generated RIN-5AH and RIN-A12 cDNAs on DNA sequencing gels revealed 31 differentially expressed bands. By Northern blot hybridization, authentic differential expression was confirmed for three cDNAs derived from RIN-5AH cells and four cDNAs from RIN-A12 cells. Nucleotide sequences were determined for these cDNAs and database searches identified one known gene that encoded heat stable antigen CD24. Of the remaining six genes, three matched with established sequence tags from fetal tissue, and three were potentially novel. By RT-PCR analysis, five of the seven genes were expressed in normal fetal and/or adult pancreas. In a detailed survey of CD24 protein expression in the pancreas using the CD24-specific monoclonal antibody J11d, CD24 was predominantly expressed in ductal epithelial cells (E13.5-15.5), developing endocrine (alpha, beta and delta) and exocrine cells (E15.5-20.5) and mature exocrine and peripheral islet delta-cells post E20.5. The retention of CD24 expression in a large proportion of delta-cells but only in a minority of alpha- and beta-cells leads us to hypothesize that CD24 may mark a pool of precursor endocrine cells within adult islets. PMID- 10365442 TI - Primary and secondary mechanisms of action of visible to near-IR radiation on cells. AB - Cytochrome c oxidase is discussed as a possible photoacceptor when cells are irradiated with monochromatic red to near-IR radiation. Four primary action mechanisms are reviewed: changes in the redox properties of the respiratory chain components following photoexcitation of their electronic states, generation of singlet oxygen, localized transient heating of absorbing chromophores, and increased superoxide anion production with subsequent increase in concentration of the product of its dismutation, H2O2. A cascade of reactions connected with alteration in cellular homeostasis parameters (pHi, [Cai], cAMP, Eh, [ATP] and some others) is considered as a photosignal transduction and amplification chain in a cell (secondary mechanisms). PMID- 10365443 TI - Endogenous porphyrin accumulation and photosensitization in the yeast Saccharomyces cerevisiae in the presence of 2,2'-dipyridyl. AB - The chelator 2,2'-dipyridyl (0.2 mM) induces a remarkable increase of protoporphyrin IX concentration as well as of its Zn-containing complex in the yeast Saccharomyces cerevisiae. Endogenous porphyrin accumulation results in five to six-fold cell sensitization to visible light (400-600 nm). Mitochondria isolated from the cells grown in the presence of 2,2'-dipyridyl accumulate protoporphyrin IX and Zn-protoporphyrin IX, while plasma membranes besides that exhibit porphyrin-type fluorescence at 670-675 nm in chloroform extract. The protoporphyrin IX content increases more than four-fold in mitochondria and two fold in plasma membranes isolated from chelator-treated cells. The relative contribution of subcellular structure photodestruction to photoinduced cell inactivation is discussed. PMID- 10365444 TI - UV-B-induced secondary conformational changes in lens alpha-crystallin. AB - The changes in turbidity and protein secondary structure of alpha-crystallin after a 72 h UV-B (302 nm) irradiation in aqueous solution have been determined by UV spectrophotometry and Fourier transform infrared (FT-IR) microspectroscopy with reflection mode. The relative transmission of alpha-crystallin aqueous solution gradually decreases with the exposure time, indicating that the transparent alpha-crystallin aqueous solution becomes opaque with prolonged UV-B irradiation. The turbidity induced by UV-B shows first-order kinetics due to the photo-induced aggregation. The modification of the secondary structure of the alpha-crystallin molecule in aqueous solution caused by this aggregation might enhance the alpha-helix and beta-turn structures from 8.14 to 14.92% and from 24.46 to 35.54%, respectively; reduce the beta-sheet structure from 60.20% to 43.77%; and leave the random coil structure almost unaltered. The secondary conformation of alpha-crystallin changes gradually but evidently with its increase of turbidity during UV-B exposure. PMID- 10365445 TI - Systemic application of photosensitizers in the chick chorioallantoic membrane (CAM) model: photodynamic response of CAM vessels and 5-aminolevulinic acid uptake kinetics by transplantable tumors. AB - The aim of this study is to modify the chick chorioallantoic membrane (CAM) model into a whole-animal tumor model for photodynamic therapy (PDT). By using intraperitoneal (i.p.) photosensitizer injection of the chick embryo, use of the CAM for PDT has been extended to include systemic delivery as well as topical application of photosensitizers. The model has been tested for its capability to mimic an animal tumor model and to serve for PDT studies by measuring drug fluorescence and PDT-induced effects. Three second-generation photosensitizers have been tested for their ability to produce photodynamic response in the chick embryo/CAM system when delivered by i.p. injection: 5-aminolevulinic acid (ALA), benzoporphyrin derivative monoacid ring A (BPD-MA), and Lutetium-texaphyrin (Lu Tex). Exposure of the CAM vasculature to the appropriate laser light results in light-dose-dependent vascular damage with all three compounds. Localization of ALA following i.p. injections in embryos, whose CAMs have been implanted with rat ovarian cancer cells to produce nodules, is determined in real time by fluorescence of the photoactive metabolite protoporphyrin IX (PpIX). Dose dependent fluorescence in the normal CAM vasculature and the tumor implants confirms the uptake of ALA from the peritoneum, systemic circulation of the drug, and its conversion to PpIX. PMID- 10365446 TI - Multiple light-harvesting II polypeptides from maize mesophyll chloroplasts are distinct gene products. AB - The major light-harvesting complex of photosystem II in higher plants is known as LHCII. It is composed of a number of chlorophyll-binding proteins sharing epitopes with each other. The number of apoproteins resolved by fully denaturing sodium dodecylsulfate polyacrylamide gel electrophoresis varies in different species. In order to know if this heterogeneity is caused by the expression of a number of homologous genes or if it is the product of post-translational modifications, we have resolved the six major apoproteins of Zea mays LHCII. Each protein is purified to homogeneity, subjected to direct protein sequencing and the sequences compared with those deduced from lhcb genes in maize and other organisms. All of the six proteins are distinct gene products, since they show differences in their primary structure. Three apoproteins are identified as products of type I lhcb genes and one each as type II and type III gene products. A sixth protein does not fit the requirements for any of the lhcb genes so far cloned and is therefore probably the product of an lhcb gene type not yet described. Our results clearly show that the major source of LHCII protein heterogeneity is the expression of many lhcb genes. Fractionation of maize LHCII by non-denaturing flat-bed isoelectric focusing resolves at least five major isoforms showing distinct differences in their polypeptide composition and also differing in their spectroscopic properties, thus suggesting that individual Lhcb gene products have distinct pigment-binding properties. PMID- 10365447 TI - Wavelength dependence of ultraviolet-induced DNA damage distribution: involvement of direct or indirect mechanisms and possible artefacts. AB - DNA damage profiles have been established in plasmid DNA using purified DNA repair enzymes and a plasmid relaxation assay, following exposure to UVC, UVB, UVA or simulated sunlight (SSL). Cyclobutane pyrimidine dimers (CPDs) are revealed as T4 endonuclease V-sensitive sites, oxidation products at purine and pyrimidine as Fpg- and Nth-sensitive sites, and abasic sites are detected by Nfo protein from Escherichia coli. CPDs are readily detected after UVA exposure, though produced 10(3) and 10(5) times less efficiently than by UVB or UVC, respectively. We demonstrate that CPDs are induced by UVA radiation and not by contaminating UVB wavelengths. Furthermore, they are produced at doses compatible with human exposure and are likely to contribute to the mutagenic specificity of UVA [E. Sage et al., Proc. Natl. Acad. Sci. USA 93 (1996) 176-180]. Oxidative damage is induced with a linear dose dependence, for each region of the solar spectrum, with the exception of oxidized pyrimidine and abasic sites, which are not detectable after UVB irradiation. The distribution of the different classes of photolesions varies markedly, depending on wavelengths. However, the unexpectedly high yield of oxidative lesions, as compared to CPDs, by UVA and SSL led us to investigate their production mechanism. An artificial formation of hydroxyl radicals is observed, which depends on the material of the sample holder used for UVA irradiation and is specific for long UV wavelengths. Our study sheds light on a possible artefact in the production of oxidative damage by UVA radiation. Meanwhile, after eliminating some potential sources of the artefact ratios of CPDs to oxidized purine of three and five upon irradiation with UVA and SSL, respectively, are still observed, whereas these ratios are about 140 and 200 after UVC and UVB irradiation. PMID- 10365448 TI - Structural characterization of the N-linked oligosaccharides from tomato fruit. AB - The primary structures of the N-linked oligosaccharides from tomato fruit (Lycopersicon esculentum) have been elucidated. For the isolation of the protein fraction, two procedures were employed alternatively: a low temperature acetone powder method and ammonium sulfate precipitation of the tomato extract. After peptic digestion, the glycopeptides were purified by cation-exchange chromatography; the oligosaccharides were released by N-glycosidase A and fluorescently labelled with 2-aminopyridine. Structural characterization was accomplished by means of two-dimensional HPLC in combination with exoglycosidase digestions and MALDI-TOF mass spectrometry. Two varieties as well as two stages of ripening were investigated. In all the samples, the same sixteen N-glycosidic structures were detected; the two most abundant glycans showed identical properties to those of the major N-linked oligosaccharides of horseradish peroxidase and pineapple stem bromelain, respectively and accounted for about 65 78% of the total glycan amount; oligomannosidic glycans occurred only in small quantities (3-9%). The majority of the N-glycans were beta 1,2-xylosylated and carried an alpha 1,3-fucose residue linked to the terminal N-acetylglucosamine. This structural element contributes to cross-reactions among non-related glycoproteins and has been shown to be an IgE-reactive determinant (Tretter, Altmann, Kubelka, Marz, & Becker, 1993). The presented study gives a possible structural explanation for reported immunological cross-reactivities between tomato and grass pollen extracts due to carbohydrate IgE epitopes (Petersen, Vieths, Aulepp, Schlaak, & Becker, 1996), thereby demonstrating the importance of the structural characterization of plant N-glycans for a more reliable interpretation of immunological data. PMID- 10365449 TI - Microbial metabolism of artemisitene. AB - Studies on the microbial transformation of the sesquiterpene endoperoxide artemisitene have revealed that artemisitene was metabolized by Aspergillus niger (NRRL 599) to yield 11-epi-artemisinin, 9 beta-hydroxydeoxy-11-epi-artemisinin and 9 beta-hydroxy-11-epi-artemisinnin. These metabolites were characterized on the basis of their spectral data. PMID- 10365451 TI - Oral and maxillofacial surgery, oral surgery and surgical dentistry: better together. PMID- 10365450 TI - Tumor promoting diterpenes from Euphorbia leuconeura L. AB - Diterpene esters of the phorbol and ingenol types are known to be highly active tumor promoting agents that typically occur in members of the Euphorbiaceae. In the present work, Euphorbia leuconeura, a rare indoor plant, is analyzed for its tumor promoting potential. Latex as well as total leaf extracts exhibited Epstein Barr-virus (EBV) inducing activity comparable to 12-O-tetradecanoyl-phorbol-13-O acetate, a well known tumor promoter. The activity of individual fractions correlated with their ingenol ester content. Three ingenol esters with EBV inducing activity could be isolated and identified. They belong to the milliamine type of diterpene esters that contain aromatic peptidyl groups. Two of them (milliamines L and M) are already known from E. milii. The third compound is identified as an isomer of milliamine F with a novel 3,20-diester arrangement. The data show a close relationship between E. leuconeura and the more popular indoor plant E. milii whose latex is also used as a powerful molluscicide. PMID- 10365452 TI - Intra-venous sedation. PMID- 10365453 TI - Sedation safety. PMID- 10365454 TI - Minimum dataset for head and neck cancer. PMID- 10365455 TI - Wisdom teeth on the waiting list. PMID- 10365456 TI - Microwave molars. PMID- 10365457 TI - Tooth surface loss. 6. Prevention and maintenance. PMID- 10365458 TI - Canalis sinuosus mimicking a periapical inflammatory lesion. AB - A case is presented in which an anatomical feature, canalis sinuosus, manifested as a periapical radiolucency on an upper canine. This may have been interpreted as an inflammatory lesion and led to the patient receiving inappropriate treatment had a further radiograph not been taken. The incisive foramen and mental foramen are well known anatomical features which may mimic periapical inflammatory lesions but it is less common for a neurovascular canal to manifest as a periapical radiolucency on an upper canine. PMID- 10365459 TI - Occupational exposure to methyl methacrylate monomer induces generalised neuropathy in a dental technician. AB - A 36-year-old dental technician for 14 years developed paraesthesia and numbness in her legs. Neurophysiological studies revealed absent sensory nerve action potentials (SNAPs) from her lower limbs and normal upper limb SNAPs on presentation. Motor nerve studies were normal. Repeat studies 2 months after leaving her job showed some improvement in the lower limb SNAPs. It is suggested that her symptoms were caused by occupational exposure to methyl methacrylate monomer. PMID- 10365460 TI - Dental composite depth of cure with halogen and blue light emitting diode technology. AB - OBJECTIVES: To test the hypothesis that a blue light emitting diode (LED) light curing unit (LCU) can produce an equal dental composite depth of cure to a halogen LCU adjusted to give an irradiance of 300 mWcm-2 and to characterise the LCU's light outputs. MATERIALS AND METHODS: Depth of cure for three popular composites was determined using a penetrometer. The Student's t test was used to analyse the depth of cure results. A power meter and a spectrometer measured the light output. RESULTS: The spectral distribution of the LCUs differed strongly. The irradiance for the LED and halogen LCUs were 290 mWcm-2 and 455 mWcm-2, when calculated from the scientific power meter measurements. The LED LCU cured all three dental composites to a significantly greater (P < 0.05) depth than the halogen LCU. CONCLUSIONS: An LED LCU with an irradiance 64% of a halogen LCU achieved a significantly greater depth of cure. The LCU's spectral distribution of emitted light should be considered in addition to irradiance as a performance indicator. LED LCUs may have a potential for use in dental practice because their performance does not significantly reduce with time as do conventional halogen LCUs. PMID- 10365461 TI - Reference doses for dental radiography. AB - OBJECTIVE: To establish reference doses for use within dental radiography. DESIGN: Retrospective analysis, single centre. SETTING: UK General Dental Practice, 1995-1998. METHOD: A statistical analysis was performed on the results from NRPB evaluations of dental x-ray equipment within general practice. The third quartile patient entrance dose was determined from 6,344 assessments of intra-oral x-ray equipment. The third quartile dose-width product was determined from 387 assessments of panoramic x-ray equipment. RESULTS: The third quartile patient entrance dose for an adult mandibular molar intra-oral radiograph is 3.9 mGy. The third quartile dose-width product for a standard adult panoramic radiograph is 66.7 mGy mm. CONCLUSION: NRPB recommends the adoption of reference doses of 4 mGy for an adult mandibular molar intra-oral radiograph and 65 mGy mm for a standard adult panoramic radiograph. These reference values can be used as a guide to accepted clinical practice. Where radiography is carried out using doses above these reference values, a thorough review of radiographic practice should be made to either improve techniques, or justify keeping the current techniques. However, attainment of doses at or below the reference values cannot be construed as achievement of optimum performance; further dose reductions below the reference value are still practicable. PMID- 10365462 TI - The relationship between water fluoridation and socioeconomic deprivation on tooth decay in 5-year-old children. AB - AIM: To examine the relationship between water fluoridation, socioeconomic deprivation and tooth decay in 5-year-olds. SETTING: 10,004 children: 1,051 in naturally fluoridated Hartlepool in 1991/92, 3,816 in fluoridated Newcastle & North Tyneside and 5,137 in non-fluoridated Salford & Trafford in 1993/94. OUTCOME MEASURES: Correlations between mean electoral ward dmft and ward Townsend Scores from the 1991 census. RESULTS: Regardless of the level of water fluoridation significant correlations were found between deprivation and tooth decay. Multiple linear regression models for dmft showed a statistically significant interaction between ward Townsend score, and both types of water fluoridation, confirming the more deprived the area the greater the reduction in tooth decay. At a Townsend score of zero (the English average) there was a predicted 43% reduction in decay in 5-year-olds in fluoridated areas. CONCLUSIONS: Tooth decay is strongly associated with social deprivation. The findings confirm that the implementation of water fluoridation has halved tooth decay in 5-year-old children and that the dental caries divide between rich and poor is reduced. PMID- 10365463 TI - A clinical teacher's experience with the Open University MA in Education. AB - In this article, I outline my experience of undertaking the Open University Master of Arts Degree in Education Management. A brief overview of the structure of this modular degree is presented. I then assess the benefits that I have gained during this course of study; I also consider the relevance of this primarily mainstream education higher qualification to those in dental education, in particular those who teach at undergraduate level and in general dental practice. PMID- 10365464 TI - [Henri Moissn, first French Nobel prize winner in chemistry: the man, the picture collector]. AB - Born in Paris in September 1852, Henri Moisson died in February 1907, two months after receiving the Nobel prize for chemistry. After a short schooling at Meaux college, he was destined to be a clock maker. He owes his vocation for chemistry to Jules Plicque, a chemist and friend at the college. Henri Moisson attended Fremy's school of chemistry at the Paris Natural History Museum and undertook pharmaceutical studies. In this presentation, we take a look at Henri Moissan's child-hood and teenage years, his scientific education and offer a glimpse of the man and the picture collector. PMID- 10365465 TI - [The scientific contributions of Moissan]. AB - Two of Moissan's most important contributions to science are presented. 1. Isolation of fluorine and the preparation of various fluorides. 2. High temperature chemistry based on the electrical are furnace. The historical and human aspects as well as the surrounding circumstances are discussed. PMID- 10365466 TI - [Fluorinated organic compounds: synthesis and biological applications]. AB - Since the discovery of molecular fluorine by Henri Moissan, fluorinated products have found numerous biological applications. Fluorine substitution can have profound effects on the properties of organic compounds. The very high electronegativity of this small substituent can modify electron distribution in the molecule, affecting its absorption, distribution and metabolism. Fluorination and 18F labeling methods are briefly described. Some examples of enzymatic inhibition by fluorinated products are examined. Nowadays, fluorine-containing medicinals are featuring significantly in such diverse areas as the provision of anti-cancer and anti-inflammatory agents, antibiotics and general anesthetics, anti-parasitic and central nervous system agents, and in cardiac therapy. Fluorocarbon emulsions show promise as blood substitutes. Several medical diagnostic techniques, such as positron emission tomography and the use of X-ray contrast agents, have profited from the use of fluorinated substances. PMID- 10365467 TI - [The electric furnace of Henri Moissan at one hundred years: connection with the electric furnace, the solar furnace, the plasma furnace?]. AB - The trace of Henri Moissan's pioneer work 100 years ago is clearly evidenced by an overview of achievements in high temperature devices; 1987: "Le four electrique" by Henri Moissan; 1948-1952: "High temperature heating in a cavity rotary kiln using focusing of solar radiation" by Felix Trombe; 1962: "The cavity rotary kiln using focused solar radiation jointly with a plasma gun" by Marc Foex; 1970: "The rotary kiln with two plasma guns and arc transfer" by Marc Foex; 1984: "The plasma furnace" by Electricite de France (EDF) at Renardieres; 1997: "The plasma furnace" by the Atomic Energy Center (CEA) at Cadarache, the VULCANO program. The first part of this contribution is devoted to Henri Moissan. Re reading his early book on the electric furnace, especially the first chapter and the sections on silica, carbon vapor and experiments performed in casting molten metal--the conclusions are outstanding--provides modern readers with an amazing insight into future developments. The last two parts are devoted to Felix Trombe and Marc Foex, tracing the evolution of high temperature cavity processus leading to the solar furnace and the present day plasma furnace at the CEA. Focus is placed on research conducted by the French National Center for Scientific Research (CNRS) with the solar and plasma furnaces at Odeillo. The relationships with Henri Moissan's early work are amazing, offering a well deserved homage to this pioneer researcher. PMID- 10365468 TI - [Utilization of a transferred arc-plasma rotating furnace to melt and found oxide mixtures at around 2000 degrees C (presentation of the film VULCANO)]. AB - Unless security measures are taken, a hypothetical accident resulting from the loss of the cooling circuit in a pressurized water nuclear reactor could cause the heart of the reactor to melt forming a bath, called the corium, mainly composed of uranium, zirconium and iron oxides as well as the structural steel. This type of situation would be similar to the Three Mile Island accident in 1979. In order to limit the consequences of such an accident, the Atomic Energy Commission has implemented a large study program [1] to improve our understanding of corium behavior and determine solutions to stabilize it and avoid its propagation outside the unit. The VULCANO installation was designed in order to perform the trials using real materials which are indispensable to study all the phenomena involved. A film on the VULCANO trials was presented at the Henri Moissan commemorative session organized by the French National Academy of Pharmacy. The rotating furnace used to melt and found the mixture simulating the corium is a direct descendant of the pioneer work by Henri Moissan. An electrical arc is directed at the center of the load to melt which is maintained against the walls by centrifugal force. After six high-temperature trials performed with compositions without uranium oxide, the first trial with real corium showed that the magma spread rather well, a result which is quite favorable for cooling. PMID- 10365469 TI - [Objectives and organization of the International Conference on Harmonization]. AB - The objective of the International Conference on Harmonization established since 1989 is to allow marketing of new drugs as soon as possible for patient benefit. At the same time harmonization decreases development time by harmonizing the content and the format of the registration file in the three regions: European Union, United States and Japan. ICH is a unique process involving health authorities and industry representatives of the three regions. The Conference which took place in July 1997 was the end of the first step of the process. At the meeting it was decided to pursue this harmonization activity, particularly in initiating a project devoted to the file format (Common Technical Document). The current period will be essentially focused on implementation of the common Guidelines and on their update in relation to scientific progress, the major part of the file now being harmonized in its content. PMID- 10365470 TI - [ICH M4: the standardized international restration dossier or "C.T.D." (common technical document)]. AB - The ICH E3 guideline "Structure and Content of Clinical Study Reports" has allowed to harmonize the structure and the contents of the study report, as well as the information to be provided in each section. Industry has wished to pursue this process and has asked for an ICH guideline regarding the complete harmonization of the registration dossier, called "C.T.D." or Common Technical Document, for the three parts of the dossier in terms of quality, safety and efficacy. A survey was carried out comparing the documentation to be provided, identifying the differences and potential obstacles to harmonization, and assessing the advantages and disadvantages of the C.T.D. The C.T.D. leading to a single registration dossier should allow pharmaceutical companies and health authorities to make savings in terms of resources (time, money and expertise), to facilitate exchanges through IT systems and to improve communication. The ICH expert groups in charge of the pharmaceutical, pre-clinical and clinical parts of the dossier are expected to start their work in February 1998. By February 2000, it is anticipated that this project will become a valuable tool for the international registration and evaluation of medicines. The C.T.D. should improve the dialogue between industry and authorities with regard to medicines and public health. PMID- 10365471 TI - [Safety of medications]. AB - Safety evaluation is an essential part for the registration of new drugs. Animal studies are performed to assess toxicity by single and repeated dose administration, potential effects on reproduction, genotoxicity, carcinogenic potential and local tolerance. Guidelines have been harmonized in the European Union since 1975, but there is no comparison in a world-side view-point, because many differences exist between the European Union, USA and Japan. This situation is very detrimental for industry, inducing the need for duplicate studies (with a waste of time and money) and to construct new files. From an ethical view point, this situation also induces overuse of laboratory animals. For all theses reasons, more or less successful harmonization of the guidelines for the safety evaluation of new products has developed since 1991. Prior to the 4th International Conference on Harmonization, important problems were still not resolved (duration of repeated dose studies in non-rodent species). Moreover, a more flexible and less complex system must be developed in the near future for better efficacy. PMID- 10365472 TI - [Contribution of explanatory notes. Efficacy of the ICH plan for international clinical development of a new drug]. AB - After the achievement of the Phase 1 of the ICH process (ICH4, Brussels, July 1997) and the recent adoption of two new Efficacy Guidelines (E5, E9) by the ICH Steering Committee, the Pharmaceutical Industry and CROs have today the necessary tools and standards of reference, to set up a Global Plan of Clinical Development for any New Molecular Entity for Human Use, within the three ICH Regions, Europe, Japan and the USA. However to achieve such a goal, the Efficacy Guidelines must officially be integrated in the Regulatory requests of each of the 3 ICH Regions ans implemented by Industry. The use of these Guidelines which concern the Clinical Development Plan and most of the content of the Clinical Documentation for Registration of New Medicinal Products, will show the qualities and defects of ICH texts, leading in the future to possible amendments, during the maintenance phase of these Guidelines (Step6 of ICH process?). The Efficacy texts cannot be separated from other ICH Guidelines concerning Quality, Safety ans above all the Multidisciplinary texts which are of great interest for the clinical development plan, registration and follow up after MAA of any New Medicinal Product. Referring to Ethnic differences between the three ICH Regions, topic E5 is the most innovative of Efficacy Guidelines for both Regional Regulatory Authorities, accepting or not foreign clinical data and International Companies, setting up Global Clinical Plan and Registration Dossier. The paper is also looking at the future impact of the other Efficacy (E10) and Multidisciplinary (M1 to M4) Guidelines which are still under consideration. PMID- 10365473 TI - [ICH4 and pharmacopoeias: similarities and differences]. AB - Since 1990, the pharmacopoeias have been regularly consulting each other on their programmes of work and have set up a Pharmacopoeial Discussion Group. This group meets regularly (twice a year) in Europe, Japan and the United States. About fifty monographs on excipients and general methods of analysis proposed by national associations of manufacturers of pharmaceutical products have been selected for convergence and harmonization among the three pharmacopoeias. The pharmacopoeias also participate in the work on the rapprochement of licensing dossiers within the framework of the cycle of international conferences known as ICH organised by the licensing authorities and manufacturers' associations of Europe, Japan and the United States. The pharmacopoeias have observer status in the QWP and Biotech WP and notably have participated in the elaboration of guidelines on analytical validation, impurities, residual solvents and specifications. If necessary, they integrate the general principles of these guidelines into their specifications. PMID- 10365474 TI - [Importance of the chemical composition and chromatographic resins since the scaling up of a procedure for the purification of proteins]. AB - Despite having the same ionic character, ion exchangers for protein separation are not identical to each other hence differences in results are generally anticipated when scaling up operations from laboratory to production. This is illustrated by comparative studies presented in this article which shows the separation of an IgG-rich fraction from sweet whey using Q-resins. One resin, available in three different particle sizes for scale changes to avoid high back pressure when increasing the size of the column, was compared with a resin that does not have the corresponding larger size particles so that scale changes have to be performed using another chemically different sorbent. Investigation of dynamic binding capacity, resolution versus loading and scale changes, demonstrated the importance of maintaining consistency in the chemical composition for an ion exchanger to eliminate differences in separation properties and therefore reduce the process development time required for specific optimization of separation conditions and maximizing productivity. PMID- 10365475 TI - [New targets in the field of antibacterial agents. Prospective aspects]. AB - Even though a great number of antibiotics and antiseptic agents are available at present, these substances are often rendered ineffective by the capacity of bacteria to develop resistance to them. The importance of continuing research in this area cannot be sufficiently stressed. Of the various strategies likely to yield innovative products in the future, two can be applied not only to the search for new antibiotics, but also for finding new antiseptics. Numerous Gram negative, as well as some Gram-positive, bacteria have efflux systems by which they excrete toxic substances, in particular, antibiotics. Because bacterial efflux pumps differ from those in eukaryotic cells, it should be possible to find inhibitors that are specific for the bacterial pumps. The colonization of biological supports by bacteria leads to the development of biofilms, which are highly resistant to both antiseptics and antibiotics. Bacteria that grow and persist within the biofilm differ metabolically from bacteria growing freely in culture. The study of the various steps in the colonization of supports might provide clues to the discovery of products that could penetrate the exopolysaccharide layer which forms the biofilm and that would be active against bacteria in this particular metabolic state. PMID- 10365476 TI - [Sexual behaviors and the HIV/AIDS risk in the general population: the methodological aspects of a national study within the context of a European Concerted Action]. PMID- 10365477 TI - Monitoring of surface microbial contamination using nitrocellulose membranes: a quantitative and qualitative study. PMID- 10365478 TI - [Bacterial contamination in a dialysis unit]. PMID- 10365479 TI - [The management of occupational exposures to biological material in the hospital environment]. PMID- 10365480 TI - [The trend in tumor mortality in man in Sardinia]. PMID- 10365481 TI - [The determination of substances of abuse: the cases of emergencies of a hospital laboratory]. PMID- 10365482 TI - [The devising of a multidimensional questionnaire for assessing the needs of cancer patients in the terminal phase]. PMID- 10365483 TI - [Basic health care and the social conditions in the neighborhoods of Cagliari]. PMID- 10365484 TI - [Managerial training as an instrument of change in health. The educational objectives and their verification in managerial training courses for the health administrators in the G. Sanelli Institute of Hygiene, 1988-1998]. PMID- 10365485 TI - Intravenous sedation. PMID- 10365487 TI - Need for registration. PMID- 10365486 TI - Sedation guidance. PMID- 10365488 TI - Digits vs dummies. PMID- 10365489 TI - Procera all-ceramic crowns: a new approach to an old problem? AB - Increasing patient expectations regarding the appearance of restorations continue to test the ingenuity and skill of the dental team. To this end the quest for sufficiently strong, metal-free, all-ceramic restorations to function in all areas of the mouth continues apace. We report on a possible solution to this problem that uses computer-aided design/computer-aided milling (CAD/CAM) technology coupled with a novel, densely sintered, ceramic material. In this article we offer suggestions for case selection, preparation design, and luting procedures, and in addition illustrate these with a number of completed cases. PMID- 10365490 TI - Marketing dental care to the reluctant patient. AB - There are many varying factors which impact on the behaviour of reluctant patients and the understanding of theoretical behavioural models can help to understand some of the key issues. This paper considers such models and the effective use of marketing communications in helping to change behaviour. The paper concludes with an acronym--OPTIMEM to help dental practitioners and practice managers plan realistic and efficient marketing communications within the overall management strategy. PMID- 10365491 TI - Tooth surface loss. 7. Dealing with failures. PMID- 10365492 TI - The psychology of dental patient care: an introduction. PMID- 10365493 TI - The psychology of dental care. 1. The common-sense approach. PMID- 10365494 TI - Fluoride ingestion from toothpaste by young children. AB - OBJECTIVE: To investigate the reported and observed brushing habits of young children and their ingestion of fluoride from toothpaste. DESIGN: In 1997, a random sample of 50 children, aged 30 months, from three districts in the North West region of England, were visited at home. METHODS: The reported and observed toothbrushing behaviour was determined and the weight of toothpaste applied to the toothbrush was measured. The amount of fluoride retained in the mouth after brushing with either a 400 ppm F or 1,450 ppm F toothpaste was determined. RESULTS: All parents claimed that their children's teeth were being brushed with a fluoride toothpaste at least once daily. The mean amount of toothpaste applied on the brush was 0.36 g of which 0.27 g (72%) was retained in the mouth. The mean amount of fluoride ingested per brushing was 0.42 mg when using the 1,450 ppm F toothpaste and 0.10 mg when using the 400 ppm F toothpaste. Although most parents applied a small amount of toothpaste a small minority applied a large amount. If using the 400 ppm F toothpaste twice daily no children of average weight would have exceeded ingestion of 0.05 mgF/kg body weight whereas 14 average weight children would have exceeded this value if using the 1450 ppm F toothpaste. CONCLUSIONS: It is essential that parents of children aged less than 7 years apply a small (pea-sized) amount of fluoride toothpaste on the toothbrush and discourage swallowing. PMID- 10365495 TI - Symptomatic sialoadenitis and sialolithiasis in the English population, an estimate of the cost of hospital treatment. AB - OBJECTIVE: To establish the annual incidence, and cost of treating, symptomatic salivary stones and sialoadenitis. MATERIALS AND METHODS: Data relating to sialolithiasis and sialoadenitis were obtained from the Department of Health with respect to the 15 health regions in England during the period 1991-1995. These were analysed to obtain the mean incidence per annum. The proportions of each condition treated on an in-patient and out-patient basis were also calculated. A survey of hospital fees was undertaken to determine the national cost for treating these two conditions. RESULTS: In the period the mean incidence of hospital admission for symptomatic sialoadenitis and sialolithiasis in the 15 health regions in England was 27.5 (19-46) and 31.5 (26-37) per million population per annum respectively. During this time there was a slight shift toward day case treatment. CONCLUSIONS: Based on hospital admission data for the period 1991-1995 the mean incidence of symptomatic sialolithiasis is relatively low, being at least 27 per million population per annum and possibly as much as 59 per million population per annum. This represents a cost to the National Health Service of up to 4,000,000 Pounds per annum. PMID- 10365496 TI - Treatment modalities in a dental fear clinic and the relation with general psychopathology and oral health variables. AB - OBJECTIVE: To assess differences among highly anxious dental patients assigned to different treatment modes (i.e. behavioural management (BM), nitrous oxide sedation (NOS), intravenous sedation (IVS). Patients were compared with regard to psychological and dental variables before treatment (e.g. number of decayed teeth), and dental variables after treatment (e.g. number of fillings made). DESIGN: Dentists experienced in the treatment of highly anxious patients allocated patients to a treatment mode based upon their clinical judgement. SETTING: Centre for Special Dental Care, Amsterdam, The Netherlands. SUBJECTS: 211 patients from this dental fear clinic. MEASURES: General psychopathology, as measured by the Symptom Checklist 90 (SCL-90), and dental anxiety (DAS, S-DAI, 10 point scale) were measured prior to treatment. From the panoramic radiograph the following pre-treatment dental variables were assessed: number of teeth, number of decayed teeth, number of retained roots, and number of root-filled teeth. After treatment, number of fillings, extractions, endodontically treated elements, number of visits, and treatment duration, were determined from the patients' records. RESULTS: Of the 144 patients who received dental treatment at the clinic, 46.5% was treated using a BM approach, 27.8% with NOS, 22.9% with IVS, and 2.8% under GA. No differences among the treatment groups were found with regard to SCL-90 and dental anxiety. The results showed that patients in the IVS group had statistically significant more decayed teeth than patients in the BM group. Furthermore, more fillings were made in the IVS group than in the BM group. CONCLUSION: Since it appeared possible to treat a large proportion of patients by BM alone, training dentists in the application of psychological methods for the treatment of anxious patients should be stimulated. In addition, future research should seek for variables that, besides oral health, are better able to discriminate between groups of highly anxious patients than measures of dental anxiety and psychopathology. PMID- 10365497 TI - A new impulse for a more comprehensive view of adolescent suicide in Europe. PMID- 10365498 TI - Another preventable jail suicide: Part Two. PMID- 10365499 TI - Letter across the Pacific. PMID- 10365500 TI - Impulsivity as a correlate of suicidal behavior in adolescent psychiatric inpatients. AB - One hundred and eighteen inpatient adolescents in a psychiatric hospital were evaluated to determine the relationship of aggression, self injury, and suicidal behavior to impulsivity. It was hypothesized that all these variables would be significantly and positively correlated with one another. This hypothesis was in part based on the results of psychobiological research that found serotonin dysfunction to be the common denominator of these psychopathological dimensions. As predicted, a significant correlation was found between the measures of suicidal behavior, aggressive behavior, and impulsivity. This correlation between suicidal behavior and impulsivity remained after partialing out the factor of aggression. Furthermore, the correlations between impulsivity and suicidality appeared greater in males than in females. Since male suicide attempters are more likely to eventually commit suicide than female suicide attempters, these findings may have a bearing on suicide prediction. PMID- 10365501 TI - Common risk factors in adolescent suicide attempters revisited. AB - The principle risk factors for suicide attempts in adolescents discussed in the literature are examined on the basis of data from a study carried out at the Geneva University Hospitals. 148 adolescents aged between 15 and 19 years form the sample. The results concerning mental disorders at the time of the suicide attempt confirm the clear predominance of affective disorder as well as the elevated frequency of comorbidity. Other main risk factors include suicidal behavior in family members and among acquaintances, poor state of health, poor integration into school or professional life, as well as sexual abuse. PMID- 10365502 TI - On suicide and attempted suicide during pregnancy. PMID- 10365503 TI - Suicide risk in high school students in Slovenia. AB - The objective of this study was to determine the psychosocial factors which differentiate suicidal adolescents from their nonsuicidal peers. By means of a specially designed questionnaire, distributed to a representative sample of 4686 Slovene high school students of both sexes aged 14-19 years, we assessed their general characteristics, suicidal ideation and behavior, family circumstances, self-appraisal of the problems and ways of solving them, engagement in sport, and exposure to suicide in their close circle. Numerous important differences were established between suicidal and nonsuicidal adolescents. The data gathered will help further research into suicidal behavior in adolescents. PMID- 10365504 TI - Substance abuse and drug-related death, suicidal ideation, and suicide: a review. AB - The high mortality rate among drug users, which is partly due to the HIV epidemic and partly due to drug-related accidental deaths and suicides, presents a major public health problem. Knowing more about prevalence, incidence, and risk factors is important for the development of rational preventive and therapeutic programs. This article attempts to give an overview of studies of the relations between substance abuse, suicidal ideation, suicide, and drug-related death. Research in this field is hampered by the absence of clear definitions, and results of studies are rarely comparable. There is, however, consensus about suicidal ideation being a risk factor for suicide attempts and suicide. Suicidal ideation is also a predictor of suicide, especially among drug users. It is correlated with an absence of family support, with the severity of the psychosocial dysfunctioning, and with multi-drug abuse, but also with requests for treatment. Every clinical examination of a drug user, not only of those who are depressed, should address the possible presence of suicidal ideation, as well as its intensity and duration. PMID- 10365505 TI - Suicide prevention in adolescents and young adults: the Geneva University Hospitals' Program. AB - In Switzerland, suicide has become the leading cause of death among the 15- to 24 year-olds. All strategies aiming at preventing suicide are most challenging, but few have demonstrated efficacy. In Geneva, we founded a Unit for Suicide Prevention as a joint venture between the Geneva University Hospitals and a charitable foundation called "Children Action." Our prevention program duplicates prevention strategies that are known to be effective: tutorials for health professionals and postvention aimed at those who have been exposed to a suicide (or suicide attempt) of a family member or a friend. But it also introduces a new strategy: a hotline for all those in the community who are in contact with a suicidal adolescent. This paper describes our experiences. PMID- 10365506 TI - Suicide attempts during adolescence: systematic hospitalization and crisis treatment. AB - This paper deals with an inpatient unit that recently opened in Geneva, specializing in the treatment of patients aged 16-21 years who had attempted suicide or felt the desire to commit suicide. This particular center was established because of the significant weaknesses found in the provision of care to adolescents who had attempted suicide. Despite the growing interest of health workers in this area of study over recent years, the frequency of suicide among the young has not decreased and there are numerous recurrences of the suicide attempts. Further, all efforts to improve the adolescents' compliance with psychiatric treatment have failed to date. The number of drop-outs from treatment is still very high. Thus, the objectives of our inpatient unit are as follows: (1) to overcome initial resistance to treatment and to improve long-term compliance; (2) to decrease the number of recurrent attempts as a consequence of the above, thus increasing life expectancy; (3) to offer the adolescents who have tried (or have contemplated) committing suicide an improved quality of life, after first helping them overcome the suicidal crisis. To achieve these goals, the therapeutic team of the unit proposes short stays during which the work with the adolescents consists of a very intensive psychoanalytic-oriented crisis intervention. Numerous practical aspects of our therapeutic approach in the inpatient unit are related here in detail, always with reference to our theoretical hypothesis. PMID- 10365507 TI - [A day in the hospital of the future]. PMID- 10365508 TI - [Vaccination of chickenpox in children with acute lymphoblastic leukaemia]. AB - Varicella vaccine has shown its efficacy to prevent the disease and complications in healthy and immunodeficient children. In this article the authors evaluate the immunologic status of acute lymphoblastic leukaemia at diagnosis and at follow up and the development of chickenpox and/or herpes zoster. Children with negative serology and continuous complete remission of acute lymphoblastic leukaemia for one year were vaccinated. Of 71 children diagnosed of acute lymphoblastic leukaemia from 1983 to 1996, 25 received the vaccine and seroconversion was obtained in 76% after one dose and 92% after the second dose. Vaccine tolerance was adequate. The incidence of herpes zoster infection was decreased in vaccinated children during chemotherapy compared to the wild-virus infected ones. Nowadays that vaccine for healthy children is recommended, we consider a priority to protect from chickenpox the children affected by leukaemia that are in continuous complete remission of the disease. PMID- 10365509 TI - [Salmonella and salmonellosis in the Asturias District, Spain, during a seven year period (1990-1996)]. AB - BACKGROUND: Retrospective study of the Salmonella sp. strains and human salmonellosis episodes registered in the Public Health Laboratory of the Principado de Asturias, Spain, over the seven year period 1990-1996. MATERIAL AND METHODS: All strains were serotyped; strains of serotypes Typhi and Typhimurium were also phage typed. The strains were grouped according to: sample of origin, age group; season of the year; time distribution, and epidemiological presentation (sporadic episode or outbreak). RESULTS AND DISCUSSION: A total of 3,255 Salmonella isolates were registered and conserved in the period under study. They were adscribed to 45 serotypes, being Enteritidis, Typhimurium, Virchow and Hadar (57.1, 26.6, 4.5 and 2%, respectively) the most frequent. 3,193 isolates were collected from clinical samples (corresponding to 3,067 patients), 32 from environmental water or sewage, 28 from food and 2 from animal stools. Relevant epidemiological findings were: the most affected age group was children under 4 years old (32.3%), the season with highest incidence was the summer. The most frequent clinical presentation was the sporadic episode of gastroenteritis (97%). Seventy five outbreaks (22 communitary and 53 family) were registered, being associated with six serotypes: Enteritidis (76%), Typhimurium, Virchow, Hadar, Infantis and Coehl. PMID- 10365510 TI - [Typification of strains of Helicobacter pylori by the detection of the cagA gene and subtypes of the vacA gene]. AB - BACKGROUND: Helicobacter pylori infection is probably the most common chronic bacterial infection in the world. The consequences of infection are pathologies like peptic ulcer, chronic gastritis, gastric cancer and gastric MALT lymphoma. The aim of this study was to detect the vacuolating cytotoxin gene (vacA) type, the cytotoxin-associated gen (cagA) of H. pylori isolates and study their association with the vacuolising activity. MATERIAL AND METHODS: Gastric biopsy specimens were obtained from dyspeptic patients. Isolates were further genotypically typed by PCR. RESULTS: All strains studied were vacA+ and 55% were cagA+. All cytotoxic strains were cagA+, subtype vacA s1/m1 and corresponded to patients with peptic ulceration. The cagA- strains (11) corresponded to subtype s2/m2. We didn't demonstrate vacuolising activity on subtypes s1/m2 and s2/m2. CONCLUSIONS: A high genetic diversity exists among strains in our environment. The subset of bacteria, vacA s1/m1/cagA+, is associated with vacuolising activity in culture cells (tox+). PMID- 10365511 TI - [Lymphocytic meningitis by mumps virus: epidemiologic, clinical, serologic and evolutive analysis of 28 cases]. AB - BACKGROUND: Mumps is a viral infection which is particularly found in children and adolescents and one of its manifestations is as lymphocytary meningitis. The aim of this study was to analyze the clinical, epidemiologic and serologic characteristics of the cases of meningitis by the mumps virus (MMV) observed during an epidemic of mumps. SUBJECTS AND METHODS: Twenty-eight cases of MMV diagnosed from December 1, 1994 to August 31, 1995 during an epidemic of mumps in the south of the province of Badajoz (Spain) were analyzed. Demographic, clinical, analytical and evolutive data were obtained. RESULTS: Cases predominated in the winter and summer in adolescents and youths (mean age 16.9 years) with a male:female relationship of 3:1. On admission most patients presented fever, headache, and parotid hypertrophy. Orchitis was observed in half of the males. No case of encephalitis was seen. Hyperproteinorrhachia was observed in the cephalorrhachidian fluid of 79% of the cases and hypoglucorrhachia was found in only two patients (7%). The course was benign, except in four patients (14%) who had sequelae (headache, unilateral hyperacusia and testicular discomfort). CONCLUSIONS: The epidemiologic and liquoral data of MMV in adolescents agree with those described in series of children. Nonetheless, the absence of encephalic involvement and the high proportion of orchitis is of note. PMID- 10365512 TI - [Validity of the notifications of red measles based on clinical diagnosis in Cataluna, Spain]. AB - OBJECTIVE: To compare the characteristics of clinical cases of measles with confirmed cases of measles registered in Cataluna during a period of four months. PATIENTS AND METHODS: An epidemiological survey was carried out on 171 cases of measles detected by a specific system of surveillance. The survey included data on age, sex, clinical symptoms, laboratory confirmation, immunization status and probable site of transmission. The relationship between the variable case (with or without laboratory confirmation) and the rest of variables was determined by the adjusted odds ratio, using a model of logistic regression. RESULTS: The epidemiological survey showed that 30.4% (52/171) had laboratory confirmation, 12.9% (22/171) had epidemiological confirmation and the rest, 56.7% (97/171) only complied with the clinical definition. The rash had a longer duration in confirmed cases (5.9 days) than in non confirmed cases (5.2 days). The lack of laboratory confirmation was associated with age younger than 5 years (ORa = 0.1; 95% CI 0.1-0.8), but there were no differences in sex, immunization status or others clinical symptoms. CONCLUSION: In clinical cases of measles the duration of the rash was lower than confirmed cases. The proportion of cases with laboratory confirmation was low (30.4%) and should be increased in the future. History of previous immunization ought not to rule out diagnosis of measles. PMID- 10365513 TI - [Calicivirus. The virus which emerges in ocean reservoirs]. PMID- 10365514 TI - [Multiple abscesses (cerebral, splenic, cutaneous) and pulmonary infection]. PMID- 10365515 TI - [Multiple pulmonary nodules and fever in a 35-year-old male]. PMID- 10365516 TI - [Regarding quality control of the Spanish Society of Infectious Diseases and Clinical Microbiology]. PMID- 10365517 TI - [Kikuchi disease in an acquired immunodeficiency syndrome patient]. PMID- 10365518 TI - [Adenovirus resistance to in vitro ganciclovir]. PMID- 10365519 TI - [Pleuro-pulmonary infection by Clostridium perfringens]. PMID- 10365520 TI - [Pyomyositis accompanied by leukemia of a myelodysplastic syndrome]. PMID- 10365521 TI - [Cervical pyomyositis and thrombosis of the internal jugular vein]. PMID- 10365522 TI - [Bacteremia by Clostridium perfringens: complication of the manual extraction of a fecaloma]. PMID- 10365523 TI - [Myocarditis by Toxoplasma in a patient with human immunodeficiency virus infection]. PMID- 10365524 TI - [Questions and answers regarding infectious endocarditis]. PMID- 10365525 TI - [Leisure-time the "best" time for tick bites. No causal FSME-therapy--Lyme borreliosis most of the time diagnosed too late]. PMID- 10365526 TI - [Centrum of interest: potency. Hormone substitution for men is topic of discussion]. PMID- 10365527 TI - [Neurogenic pain after deafferentation through trauma. Little-known pain syndrome following amputation, brachial plexus injury and nerve root avulsion]. AB - Amputations may be followed by such phenomena as phantom-limb pain, and pain and involuntary movements of the stump. The sequelae of brachial plexus injuries or cervical root avulsion--the second large group of deafferentiation lesions associated with neurogenic pain--are less varied, and most of these cases involve damage to the substantia gelatinosa of the spinal cord. The two groups of deafferentiation lesions are described on the basis of our own experience and reports in the literature. A generally applicable effective form of treatment is not known, and possible therapeutic approaches, which need to be adapted individually, are discussed. PMID- 10365528 TI - [Neuralgic neuropathies--complex pathogenesis and therapy. Trigeminal neuralgia- neuralgic shoulder amyotrophy--post-herpetic neuralgia]. AB - A wide spectrum of manifestations and causes makes the diagnosis of neuralgic neuropathies difficult. In most cases, however, clinical and/or electrophysiological signs of a peripheral nerve disorder can be established. The pathomechanisms underlying pain are complex and not fully understood. They are not limited to the site of a lesion, but encompass the entire peripheral neurone, and may even involve the central neurones. Pathobiological correlates of chronic pain have been identified to involve the entire pain-mediating neuronal pathway. PMID- 10365529 TI - [Pain therapy in diabetic neuropathies]. PMID- 10365530 TI - [Sexually transmitted parasitoses and mycoses]. PMID- 10365531 TI - [Spondylarthritis. 5. (final): therapy of special manifestation forms of spondylarthritis]. PMID- 10365532 TI - [Therapy of recurrent exudative erythema multiforme. Effectiveness of thalidomide -report of a case]. AB - The authors report on the efficacy of thalidomide used to treat recurrent erythema exsudativum multiforme associated with herpes simplex with massive involvement of the entire skin and also the nasal mucosa in a 73-year-old woman. Efficacy in terms of healing and the prevention of recurrence proved to be very good. PMID- 10365533 TI - Allogeneic stem cell transplantation for patients with high-risk myelodysplastic syndrome. AB - Allogeneic stem cell transplantation (allo-SCT) is the only treatment with curative potential for patients with myelodysplastic syndrome (MDS). From June 1986 to April 1997, we treated 12 patients with primary MDS (5 men, 7 women, median age, 36.5 years) by allo-SCT. All patients had one or more of the following poor prognostic factors: intermediate-2 or high-risk categories according to the International Prognostic Scoring System; disease progression during follow-up; heavy transfusion requirements and recurrent infections. The median duration from diagnosis of MDS to allo-SCT was 6 months. The preconditioning regimen included total body irradiation combined with either high dose cytarabine (n = 6), high-dose cyclophosphamide (n = 4), or other regimens (n = 2). Ten patients received bone marrow transplantations and two patients received peripheral blood stem cell transplantations. Prophylaxis for graft versus-host disease (GVHD) consisted of standard cyclosporin and short-course methotrexate. Acute GVHD of grade 2 or above occurred in 10 patients, while chronic GVHD occurred in seven of the nine patients who survived longer than 6 months after allo-SCT. With a median follow-up of 50 months, all nine patients with human leukocyte antigen (HLA)-matched sibling donors survived. One patient had a relapse 6 months after transplantation and achieved complete remission again with low-dose cytarabine therapy. The three patients receiving allo-SCT from unrelated or HLA-mismatched donors died of grade 3 to 4 acute GVHD and infection within 5 months after transplantation. The estimated disease-free survival at 4 years was 67% (95% confidence interval, 40-93%), and the overall survival was 75% (95% confidence interval, 50-99%). Our data suggest that allo SCT should be considered early in the clinical course for young MDS patients with a poor prognosis and a matched sibling donor. PMID- 10365534 TI - Cardiorespiratory response of heart transplantation recipients to exercise in the early postoperative period. AB - In this study, we evaluated the cardiorespiratory function of orthotopic heart transplantation (OHT) recipients during exercise. Seventeen male OHT recipients, ranging in age from 22 to 60 years, participated in this study 47 +/- 21 days after surgery. The control group consisted of 17 sedentary healthy men. Breath-by breath measurement of cardiorespiratory function was obtained during the incremental exercise of leg cycling. At peak exercise, the oxygen (O2) uptake (16.5 +/- 3.3 vs 33.9 +/- 8.2 mL.kg-1.min-1), work rate (82 +/- 19 vs 169 +/- 42 watts), heart rate (HR), O2 pulse, and blood lactate level of the OHT recipients were significantly lower than the respective values of the control group. At the ventilatory threshold, the OHT group also showed a significantly lower O2 uptake (10.7 +/- 1.6 vs 18.3 +/- 5.1 mL.kg-1.min-1), work rate (39 +/- 12 vs 89 +/- 33 watts), HR, O2 pulse, ventilatory equivalent for O2, and ventilatory equivalent for carbon dioxide. The OHT recipients showed a high resting HR (97 +/- 7 beats/min) and a low peak HR (123 +/- 14 beats/min) during exercise, and their HR continued to increase for 1 to 3 minutes after cessation of exercise. Our data revealed a low level of cardiorespiratory endurance in OHT recipients during the early postoperative stage. A multidisciplinary cardiac rehabilitation program should be considered to enhance physical functional capacity and quality of life, and promote return to work. PMID- 10365535 TI - Fertilization capability of frozen epididymal sperm for intracytoplasmic sperm injection. AB - Both microsurgical epididymal sperm aspiration (MESA) and intracytoplasmic sperm injection (ICSI) are great advances in assisted reproductive techniques. By using the ICSI technique, frozen sperm from the epididymis can result in successful fertilization. The epididymal sperm retrieved via MESA can be cryopreserved for an in-vitro fertilization (IVF) procedure, thus, making repeat surgical retrieval of sperm unnecessary. We report a retrospective analysis of 24 ICSI cycles in 16 patients with obstructive or nonreconstructable azoospermia. Fresh epididymal sperm was used in 13 ICSI cycles and frozen-thawed epididymal sperm was used in the other 11. We compared the fertilization capability of ICSI using frozen thawed epididymal sperm with fresh epididymal sperm. Eleven patients became pregnant and five of these pregnancies resulted from frozen epididymal sperm. The fertilization rate per oocyte was 58% with fresh sperm, and 66% with frozen thawed sperm. The rate of clinical pregnancy for one embryo transfer was 46% with fresh sperm, and 45% with frozen-thawed sperm. There were no significant differences between fresh and frozen-thawed spermatozoa in the fertilization rate of oocytes or the clinical pregnancy rate. Our results suggest that we should cryopreserve supernumerary spermatozoa during a MESA/ICSI procedure in order to avoid repeated scrotal surgery. PMID- 10365536 TI - The Ilizarov technique for treatment of sequelae of childhood-acquired bone and joint infection. AB - Bone and joint infections in childhood can result in various sequelae including deformity and limb length discrepancy (LLD). Management of these sequelae is difficult and must be individualized. In this study, we retrospectively examined the efficacy of treatment with Ilizarov techniques in 30 patients suffering from sequelae of bone and joint infections in childhood, treated from 1989 to 1994. These cases comprised 17 hip infections, two septic shoulders, and 11 cases of osteomyelitis (4 femurs, 5 tibiae, 1 humerus, and 1 forearm). All patients had some evidence of LLD. There were 13 cases of hip deformity and 10 cases of unstable hips. There were also 10 cases of angular deformity and five of nonunion of bone. The mean age at treatment was 21 years. Twenty-eight patients underwent Ilizarov lengthening procedures and 18 of them underwent deformity or instability correction simultaneously. Two patients underwent Ilizarov deformity correction only. Various techniques were used, including Schanz osteotomy plus mid-shaft femoral lengthening and distraction callotasis with and without an intramedullary nail. Postoperative complications included stiffness of joints in six patients, pin tract infection in six, fracture in five, malunion in five, nonunion in four, callus shortening in four, nerve palsy in two, and over-lengthening in one. The mean duration of follow-up was 72 months. The average length gain was 6.6 cm in shortened bones, with a mean external fixator index of 40.5 days/cm. All patients were satisfied with the functional and cosmetic results. We conclude that Ilizarov techniques are effective in treating sequelae of childhood infections of bones and joints. PMID- 10365537 TI - Surgical treatment of severe perthes disease: comparison of triple osteotomy and shelf augmentation. AB - The optimal management of the severe form of Perthes disease is controversial. This retrospective study evaluated the results of two procedures in two groups of patients with Perthes disease. The Catterall classification was adopted for grouping of patients before treatment. The Herring classification was used for comparison of the follow-up radiographs. Under the concept of surgical containment, triple innominate osteotomy was performed in 14 patients at an average age of 8 years 7 months with a mean follow-up period of 4 years 3 months. Staheli's shelf augmentation was performed in 14 patients at an average age of 10 years 2 months with a follow-up period of 3 years 8 months. Radiologically, femoral head subluxation, acetabular coverage, acetabular angle, and center-edge angle were markedly improved in both groups. In the clinical evaluation using modified Sundt's criteria, both procedures were effective. Satisfactory results were achieved in 79% of 14 patients (5 good, 6 fair, and 3 poor) in the triple osteotomy group, and 100% of 14 patients (two good, 12 fair) in the shelf augmentation group. Nevertheless, triple innominate osteotomy is technically more demanding with a longer operating time and resulted in more complications. Staheli's shelf augmentation is a simpler procedure with better coverage of the acetabulum. However, asphericity of the femoral heads was observed more frequently in this group. The Herring lateral pillar classification was better than the Catterall classification in predicting the final outcomes in this study. PMID- 10365538 TI - Risk factors for postoperative femoral fracture in cementless hip arthroplasty. AB - The objective of this study was to evaluate the incidence of and risk factors for femoral fracture in patients who underwent cementless hip arthroplasty during a 3 year period. Several predisposing factors have been reported; we tried to find another predictive indicator that could be recognized preoperatively. The records of all patients who underwent cementless hip arthroplasty from December 1993 to December 1996 were reviewed. The characteristics and clinical features (including age, gender, diagnosis, geometry of the proximal femur, and quality of bone) of the patients who had fractures were compared with those of patients who did not have fractures. During the 3-year study, 425 patients underwent a total of 454 cementless hip arthroplasties. There were 16 postoperative fractures (3.5%, 16 patients). Patients who suffered femoral fracture were significantly older than patients without fracture (65.6 +/- 10.9 yr vs 52.6 +/- 16.2 yr, p < 0.001). The fracture group had poorer preoperative bone quality compared with the nonfracture group (3.3 +/- 0.6 vs 3.8 +/- 0.7, Singh's Index of Osteoporosis, p < 0.01). The canal flare index of the proximal femur was significantly lower in the fracture group than in the nonfracture group (3.3 +/- 0.40 vs 3.8 +/- 0.7, p < 0.01). Our results indicate that old age and osteoporosis affect the likelihood of periprosthetic femoral fractures, and that a low flare index is a predictive indicator of femoral fracture. These factors should be taken into account during preoperative planning, and cemented arthroplasty should be considered for patients with these risk factors. PMID- 10365539 TI - Ultrasound-guided percutaneous transthoracic needle aspiration biopsy for diagnosis of pulmonary lesions in advanced HIV infection. AB - Pulmonary diseases remain the most common complication associated with high morbidity and mortality in patients with human immunodeficiency virus (HIV) infection. Invasive diagnostic procedures are often needed to establish a specific diagnosis of pulmonary disease. We report our experience with ultrasound (US)-guided percutaneous transthoracic needle aspiration (PTNA) biopsy in 20 consecutive patients with advanced HIV infection who presented with a variety of pulmonary lesions with or without pleural effusion. A specific diagnosis was established in 16 patients (80%), with infection being the most common etiology. Sputum culture yielded the same causative pathogen in three patients (15%) and all had more than one bacterial or fungal isolates. Mild pneumothorax, the only complication, was observed in two patients (10%) following the procedure. Neither patient required chest tube drainage. Our findings suggest that US-guided PTNA can be a useful and safe alternative to fluoroscopy-guided PTNA in selected HIV infected patients with focal pulmonary lesions and pleural effusion. PMID- 10365540 TI - Displacement effect of valproate on bilirubin-albumin binding in human plasma. AB - Drugs that can displace bilirubin from plasma albumin binding may cause various late manifestations of brain damage if given to newborns with unconjugated hyperbilirubinemia. The purpose of this study was to examine the displacement effect of valproate (VPA) on bilirubin-albumin binding in human serum albumin (HSA) and human plasma. Bilirubin-HSA solution and bilirubin-plasma solution containing no or serial concentrations of VPA were prepared, and free bilirubin in these solutions was measured by the enzyme oxidation method. VPA displaced bilirubin from bound bilirubin-albumin. The binding constant (KD) of VPA to the high affinity bilirubin-binding site of HSA and plasma protein were 11.6 L/mmol and 9.0 L/mmol respectively. The displacement effect may occur at the clinical concentration range of VPA, with a maximal displacing factor of 1.5 to 4.7. These results suggest that the clinician should use VPA in jaundiced neonates with caution. PMID- 10365541 TI - Muscle phosphofructokinase deficiency (Tarui's disease): report of a case. AB - A 14-year-old girl had an acute episode of rhabdomyolysis after vigorous exercise and seizures. Laboratory studies revealed elevated creatine phosphokinase (CPK) activity and myoglobinuria without acute renal failure, as well as mild indirect hyperbilirubinemia, and hyperuricemia. The elevated CPK activity, mild indirect hyperbilirubinemia, and hyperuricemia persisted during a 10-month follow-up period, during which chronic hemolysis without overt anemia was also noted. A muscle biopsy specimen from the left biceps muscle revealed occasional muscle fiber necrosis and mild excess of glycogen accumulation on periodic acid-Schiff staining. Histochemical reactions were negative with phosphofructokinase (PFK) stain when fructose-6-phosphate was used as the substrate, but positive when fructose 1,6-bisphosphate was used as the substrate. These findings confirmed the diagnosis of muscle PFK deficiency (Tarui's disease), which is a defect of glycolysis in muscles and erythrocytes. Less than 40 such patients have been reported to date. When a specific metabolic myopathy is suspected in children with rhabdomyolysis, symptoms of hemolysis should also be sought to identify Tarui's disease. To the best of our knowledge, this is the first case of Tarui's disease identified in Taiwan. PMID- 10365542 TI - Primary sclerosing cholangitis in a child. AB - Primary sclerosing cholangitis (PSC) is a rare disease in Taiwan and has not been described in Taiwanese children previously. We report a 4-year-old girl who presented with prolonged fever, eosinophilia (11%), hepatomegaly, and markedly elevated serum levels of alkaline phosphatase (3,318 IU/L) and gamma-glutamyl transpeptidase (475 IU/L). Subsequent investigations including endoscopic retrograde cholangiopancreatography and liver histology confirmed the diagnosis fo PSC. Treatment with a low dose of prednisolone for 2 months and ursodeoxycholic acid during 32 months of follow-up resulted in clinical remission and halted disease progression. A high index of suspicion is necessary for physicians to diagnose this disorder in children with chronic liver disease. Our experience in this case indicates that therapy with prednisolone and ursodeoxycholic acid may be helpful for the treatment of PSC in children, and suggests the need for more trials of combined therapy. PMID- 10365543 TI - Acinetobacter meningitis: four nosocomial cases. AB - We report the clinical features and therapeutic outcomes of four patients with multiantibiotic-resistant Acinetobacter meningitis. There were three males and one female, aged from 17 to 49 years. Three of them had suffered from head injuries with skull fractures, and the other suffered from an intracerebral hemorrhage and underwent a craniotomy. All four patients acquired nosocomial Acinetobacter meningitis, and multiantibiotic resistance developed. After treatment with imipenem/cilastatin, three of the four patients survived; one died of multiorgan failure. Because the clinical manifestations of Acinetobacter meningitis are similar to those of other gram-negative bacillary meningitis, the diagnosis can only be confirmed by bacterial culture. Resistance to multiple antibiotics, including third-generation cephalosporins, is frequently seen in patients with nosocomial Acinetobacter meningitis, and imipenem/cilastatin seems to be the antibiotic of choice for this potentially fatal central nervous system infection. PMID- 10365544 TI - Intraspinal osteogenic meningioma: report of a case. AB - Dense calcification and psammomatous bodies are common in spinal meningioma, but are rarely reported in osteogenic meningioma. We present a 73-year-old woman with an extramedullary, intradural tumor located at the T5 vertebra. The tumor showed mixed intensity and heterogeneous enhancement on the T1-weighted image and hypointensity on the T2-weighted image, and was situated near the spinal nerve root. The tumor's initial symptom was myelopathy, as is usual with spinal meningioma. We successfully removed the tumor under microscopy and found it to be separated from the vertebral column by the epidural space. The symptoms and signs improved gradually after the operation. Because the pathologic examination revealed areas of lamellar bone with bone marrow in the transitional meningioma, and because these were not related to the psammomatous bodies, osteogenic meningioma was diagnosed. Metaplasia of arachnoid cells is considered to be the putative etiology of osteogenic meningioma. PMID- 10365545 TI - Racism in medicine. PMID- 10365546 TI - The recruitment of breast cancer survivors into cancer control studies: a focus on African-American women. AB - The recruitment of African Americans into cancer prevention and control studies has presented a major challenge to scientific investigators. Scientific findings, whether biomedical or behavioral, may not be appropriate and applicable to ethnic minority populations unless they are adequately represented as study participants. Moreover, the need to involve greater numbers of ethnic minorities is quite urgent due to the poor morbidity and mortality outcomes associated with ethnic minority group membership. Such is the case with breast cancer survivorship. The purpose of the study was to test a personalized recruitment strategy on response rate in African-American women. The response rate of 45% (n = 117) African Americans and 64% (n = 161) white subjects indicated only limited success in the recruitment of the African-American breast cancer survivors. The recruitment result suggests that culturally relevant recruitment strategies (e.g., inclusion of African-American investigators, culturally consistent letter of recruitment) may be insufficient in adequately increasing research participation. Therefore, further studies on investigating factors that influence research participation (eg, type of incentives, and schedule of payment as well as type of stationery and stamps used) are needed. PMID- 10365547 TI - Prolactin response to the severity of surgical insult. AB - Prolactin levels are elevated significantly during the recovery process from surgical insult, implying a role for prolactin in the neuroendocrine immune network. This study examined the importance of severity of surgical insult to the prolactin response. Two groups of surgical patients were chosen consecutively and studied prospectively. Seven patients scheduled for "clean" elective surgery, i.e., herniorrhaphy and laparoscopic cholecystectomy, were compared with seven patients scheduled for prolonged abdominal exploration. Blood was drawn for prolactin and cortisol at 8:00 AM on the day of surgery and on postoperative days one, three, and five. Using a two-tailed test, preoperative prolactin levels and levels on postoperative days three and five were significantly different in the prolonged surgery group (.012 and .002, respectively). There were no statistically significant differences in preoperative and postoperative prolactin levels in the clean surgery group. Cortisol levels were not significantly elevated in either group. These results indicate that the prolactin response to surgery is related to the severity of the surgical insult. PMID- 10365548 TI - Prevalence of disabling conditions among African-American children and youth. AB - This article reports on the prevalence of disabling conditions among children and youth in the African-American subpopulation. The health status of the African American population as a whole is discussed as well as the disabling conditions among African-American children and youth specifically. The unique social, economic, and health conditions relative to African-American children and youth are highlighted. Recommendations for future research, policy, and practice are made to alleviate problems surrounding African-American children and their families. PMID- 10365549 TI - Predictors of cesarean section delivery among college-educated black and white women, Davidson County, Tennessee, 1990-1994. AB - Cesarean section delivery increases the cost, morbidity, and mortality of childbirth. Cesarean section rates vary nationwide with the highest rates occurring in the southern United States. The Department of Health and Human Services has published year 2000 objectives that include a 15% reduction in the cesarean section rate. This study identified factors contributing to cesarean section delivery among a cohort of college-educated black and white women in Davidson County, TN. Logistic regression models were applied to Linked Infant Birth and Death certificate data from 1990-1994. Data on singleton first births for 606 black women and 3661 white women completing 16 years of education were analyzed. College-educated African Americans were at a significantly higher risk of cesarean section delivery than whites. This difference could not be accounted for by controlling for all other variables. The geographic differences in cesarean section rates in this country may be the result of varying in provider practice styles, perceptions, or attitudes. Improving the health of women and children will require establishing a system of maternity care that is comprehensive, case-managed, culturally appropriate, and available to all women. PMID- 10365550 TI - Kenney Memorial Hospital. PMID- 10365551 TI - Spontaneous spinal subdural hematoma in a young adult with hemophilia. AB - Spontaneous spinal subdural hemorrhage is a rare clinical problem that usually manifests with a sudden onset of pain and paralysis. This article reports on an 18-year-old male with hemophilia A and cerebral palsy, who experienced a several month history of transient back, hip, and leg pain accompanied by gait difficulties that ultimately culminated in a more striking episode of lower extremity weakness, irritability, and diffuse pain involving the neck, back, and legs. In the absence of any clinical or radiographic evidence of hemarthrosis, osteomyelitis, or intracranial hemorrhage, imaging of the spine disclosed a large, apparently multicompartmentalized intraspinal lesion, consistent with old hemorrhage. This extended from the thoracic to the sacral region, with the largest extent at the lumbosacral junction. Following correction of factor VIII levels, surgical exploration was undertaken and demonstrated liquefied blood within the subdural space without violation of the underlying arachnoid. Because the chronic subdural blood flowed quite easily through the dural opening by simply angling the operating table, a limited exposure was required to achieve a substantial evacuation of the clot. This case calls attention to the often protean manifestations of this process, the potential for a chronic course to the clinical symptoms, and the possibility of achieving substantial clot evacuation and clinical recovery with a limited operative approach. PMID- 10365552 TI - [New antibiotics in the pipeline]. PMID- 10365553 TI - [COMT inhibitors]. PMID- 10365554 TI - [Toxoplasmosis]. PMID- 10365555 TI - [Secondary vegetables. Greens and legumes]. PMID- 10365556 TI - [Infant botulism--rare, but dangerous. Preventive measures: infants must not be given honey!]. PMID- 10365557 TI - The eosinophil, a Trojan Horse? PMID- 10365558 TI - Sleep related breathing disorder: is it sleep apnea syndrome? PMID- 10365559 TI - On sleep and death: cardiovascular risk of the obstructive sleep apnea syndrome. PMID- 10365560 TI - Chronic eosinophilic pneumonia with pleural effusion. AB - The case history of a 77-year-old lady with chronic eosinophilic pneumonia is presented. The diagnosis was difficult due to the simultaneous presence of a pleural effusion and congestive heart failure. Radiological findings and treatment are discussed. PMID- 10365561 TI - Acute abdominal pain and eosinophilia, two cases of eosinophilic gastroenteritis. AB - Two patients are presented who were admitted with acute abdominal pain for which they underwent laparotomy. No clear-cut diagnosis could be established during operation. Eventually, eosinophilic gastroenteritis was diagnosed and treated with corticosteroids. The heterogeneous presentation of eosinophilic gastroenteritis is discussed, ranging from mild non-specific gastrointestinal symptoms to an acute abdominal emergency prompting surgical intervention. The pathogenesis and treatment of eosinophilic gastroenteritis are discussed. PMID- 10365562 TI - Prevalence and severity of sleep disordered breathing in a group of morbidly obese patients. AB - BACKGROUND: Obesity may be complicated by sleep disordered breathing (SDB). The presence of SDB is associated with increased morbidity and mortality. Patient characteristics, pulmonary function tests and daytime arterial blood gas analyses may help to identify patients with SDB. These variables and the prevalence and severity of sleep disordered breathing were studied in a group morbidly obese patients. METHODS: Forty-eight patients, 19 men and 29 women who were referred to our clinic of internal medicine because of their obesity were included. Characteristics, pulmonary function tests and daytime arterial blood gas analyses of groups with different grades of SDB were compared. RESULTS: Male subjects had significantly more apnoeas/hypopnoeas per hour (AHI) (18.4 +/- 20.9 versus 4.8 +/ 9.4) with more desaturation, a lower mean saturation (92.6 +/- 4.1 versus 96.1 +/- 1.6) and a lower saturation nadir (73.8 +/- 12.0 versus 83.1 +/- 6.7). Five (26%) of the male subjects and none of the female subjects had severe SDB (AHI > or = 25). Subjects were divided into three groups according to the severity of their SDB: twenty-nine subjects (23 women and 6 men) with AHI < 5, 14 subjects (6 women and 8 men) with AHI > or = 5 and < 25 and 5 subjects, all men, with AHI > or = 25. Except for gender no significant differences were found between the three groups. CONCLUSION: Our study confirms the findings that morbidity obese men have SDB more frequently and more severely than obese women. Patient characteristics other than gender, pulmonary function tests and daytime blood gas analyses have no predictive value. PMID- 10365563 TI - Sleep complaints and sleep disordered breathing in hemodialysis patients. AB - BACKGROUND: The objective of the study was to determine the prevalence of sleep complaints and of sleep disordered breathing (SDB) in hemodialysis patients not selected for sleep complaints and to determine the effect of hemodialysis on SDB. The feasibility of home recording of sleep related respiration in these patients was also studied. METHODS: The patients completed a questionnaire and parameters of SDB were examined in the home setting on nights following dialysis and nights following no dialysis with the Edentrace II Recording System. RESULTS: Six (46%) of 13 patients had sleep complaints. Symptoms suggestive for sleep apnea syndrome were found in four (31%) of these 13 patients. In three (75%) of these four patients SDB was found. Sleep related respiration was monitored in 15 patients. Registrations satisfactory for interpretation were obtained in all patients. SDB was observed in five (33%) of these 15 patients. There were no significant differences in parameters of SDB between nights following dialysis and nights following no dialysis. CONCLUSIONS: Home recording of sleep related respiration in hemodialysis patients is feasible. Sleep complaints and SDB are common in these patients. No clinically significant differences in SDB were found between nights following dialysis and nights following no dialysis. PMID- 10365564 TI - Can birth defects be prevented? PMID- 10365565 TI - Birth defects recognized in 10,000 babies born consecutively in Port Moresby General Hospital, Papua New Guinea. AB - A daily record was made of defects recognizable at birth or soon afterwards in 10,000 babies born consecutively at Port Moresby General Hospital, Papua New Guinea, between January 1985 and May 1986. The overall prevalence of birth defects in this series was 1.16%. All of the affected babies were singletons, 27% presented with multiple defects, and 14% were stillborn. There was a predominance of male babies in the series as a whole and more particularly in the group of affected babies (female:male ratios 1:1.15 and 1:1.50 respectively). The parts of the body most commonly affected were the limbs, head and neck, and central nervous system. The majority of the mothers originated from provinces neighbouring Port Moresby, although all the provinces were represented. Defects were more common in babies of mothers from island provinces (1.9%) than from highland (1.1%) or coastal/lowland (1.0%) provinces. The mean birthweight for all the babies in the series for whom records were available was 3.03 kg (SD 0.57), and for the abnormal babies 2.86 kg (SD 0.70). The highest mean birthweight was recorded for babies of highland mothers and the lowest for babies of coastal/lowland mothers. PMID- 10365566 TI - Health impact assessments of malaria and Ross River virus infection in the Southern Highlands Province of Papua New Guinea. AB - Malaria at an elevation of 1050 metres is common and highly endemic in the Tagari Valley in the Southern Highlands of Papua New Guinea. Health impact assessments showed that the risks of malaria and epidemic polyarthritis at a gasfield development project in this area were high. Baseline malariometric surveys were conducted in four villages in June and August 1990 and two follow-up surveys (May and December 1991) were made in the village of Nogolitogo near the gasfield pioneer base camp. A total of 941 blood smears were examined. Average malaria prevalence rates decreased with altitude from 56% (at 1050 m) to 9% (at 1700 m) for children 1-9 years of age and from 45% (at 1050 m) to 8% (at 1550 m) for those aged 10 years or more. The spleen rate for children less than 10 years old did not vary significantly with altitude, but average enlarged spleen for all ages decreased with altitude. Mean packed cell volume increased with altitude. Plasmodium falciparum was the most common malaria parasite found and Anopheles punctulatus the predominant vector. Ross River arbovirus (RRV) antibody prevalence was 59%. These results indicate frequent or constant transmission of malaria and pathogenic arboviruses. Entomological and epidemiological data suggested that the vulnerability of the valley community, the receptivity of the environment and the health hazards from malaria and RRV were high. Nonimmune Papua New Guineans and expatriate employees face high health hazards; therefore effective preventive measures are required to mitigate epidemics and avoid the likely heightened transmission of malaria and arboviruses caused by the development project. PMID- 10365567 TI - A survey of under-18 year old and 20-29 year old primigravidae delivered at the Port Moresby General Hospital: a comparative study of their sociodemographic and sexuality characteristics and contraceptive knowledge and experience. AB - From July 1992 to August 1993, 330 under-18 year old primigravidae (cases) and 330 randomly selected 20-29 year old primigravidae (controls) who were delivered at the Port Moresby General Hospital were sequentially studied, using a standardized, pretested, precoded questionnaire. In stepwise logistic regression analysis, significantly more of the cases had menarche at less than 15 years of age, learned before menarche that sex causes pregnancy, were of highland origin, were unemployed, or had partners who were unemployed; significantly fewer of the cases thought that one sexual act could cause pregnancy, had knowledge of or had ever used a family planning method, or had planned this pregnancy. PMID- 10365568 TI - Review of 17 cases of ectopic pregnancy at the Vila Central Hospital in Vanuatu. AB - A review of cases of ectopic pregnancy operated upon at Vila Central Hospital during 1992 with an analysis of clinical presenting features and diagnostic factors is presented. Comparison is made between hospital, regional and national figures and possible explanations for the differences are given. Recommendations are made to ensure that ectopic pregnancy is always at the forefront of differential diagnosis in women presenting with abdominal pain. PMID- 10365569 TI - A case of advanced viable extrauterine pregnancy. AB - Advanced extrauterine pregnancy with a successful outcome is a rare event. A case is presented of a 34-year-old woman at 35 weeks gestation whose abdominal pregnancy was successfully managed. The diagnostic and management problems associated with abdominal pregnancy are discussed, and especially the controversial issues of the treatment of the placenta after delivery. The reasons for the high maternal and perinatal mortality associated with the condition are analyzed. PMID- 10365570 TI - Tissue inhibitor of metalloproteinases-1 and matrix metalloproteinase-3 in Japanese healthy children and in Kawasaki disease and their clinical usefulness in juvenile rheumatoid arthritis. AB - BACKGROUND AND METHODS: To determine the clinical values of tissue inhibitor of metalloproteinases-1 (TIMP-1) and matrix metalloproteinase-3 (MMP-3) in juvenile rheumatoid arthritis (JRA), we measured serum levels of these enzymes with rapid one-step sandwich enzyme immunoassay. Forty-one JRA patients, 48 normal healthy children (NC) and 10 Kawasaki disease (KD) patients were investigated. RESULTS: Serum TIMP-1 levels in NC corresponded to those in normal adults reported in the literature, while MMP-3 levels were lower than those in healthy children and the ratio of MMP-3/TIMP-1 decreased. The TIMP-1 levels in JRA and KD at the first clinic examination were statistically higher than those in NC (P < 0.05) and MMP 3 levels and MMP-3/TIMP-1 in JRA were significantly higher than those in NC (P < 0.0001 and 0.0005, respectively) and KD (P < 0.001 and 0.0005, respectively). In JRA, MMP-3 levels of patients with arthritis were statistically higher than those of patients without arthritis (P < 0.05) and MMP-3 levels were correlated with C reactive protein (rs = 0.465, P < 0.05), while TIMP-1 did not (rs = 0.340). There was a positive correlation between serum levels of MMP-3 and TIMP-1 and prognosis (rs = 0.733, P < 0.05). CONCLUSION: In JRA, the serum MMP-3 level is a useful marker to evaluate joint damage, while serum TIMP-1 remains an acute phase reactant. PMID- 10365571 TI - Serum soluble L-selectin in childhood acute lymphoblastic leukemia. AB - BACKGROUND: Determination of the serum level of soluble (s)L-selectin has been advocated for monitoring patient response to treatment in leukemia. The aim of the present study was to find out whether serum levels of sL-selectin correlated with treatment for acute lymphoblastic leukemia (ALL) in children. METHODS AND RESULTS: Serum samples were obtained from 30 children with ALL, either newly diagnosed during induction therapy, in remission, in maintenance therapy, at the end of treatment or after relapse. Levels of sL-selectin were assayed in the serum of children during the clinical course of ALL using the sandwich enzyme linked immunoabsorbent assay. Serum sL-selectin concentrations decreased significantly from diagnosis to the end of intensive chemotherapy in children with ALL and increased in the time of relapse. CONCLUSION: These results suggest that monitoring of sL-selectin may be useful for evaluating leukemia activity. PMID- 10365572 TI - Serum lipoproteins and apolipoprotein E in infants with congenital hypothyroidism. AB - BACKGROUND: Hypothyroid adults have a high risk of atherosclerosis, secondary to increased levels of various cholesterol fractions, particularly low-density lipoprotein cholesterol (LDL-C). We investigated the existence of a correlation between thyroid hormone deficiency and serum lipoproteins and a possible effect of different apolipoprotein E (apoE) phenotypes on lipoprotein levels in 75 infants with hypothyroidism. METHODS: Seventy-three of the 75 infants had congenital hypothyroidism. At the age of one month, prior to the initiation of thyroid hormone substitution therapy, thyroid-stimulating hormone (TSH), thyroid hormones and lipid profile parameters were determined. Subsequently, apoE phenotyping in all patients was performed by isoelectric focusing followed by immunoblotting. RESULTS: Significant negative correlations were identified between triiodothyronine (T3) and LDL-C and total cholesterol (TC) levels and between thyroxine (T4) and TC levels. There were no correlations between TSH and free (F)T4 and lipid profile parameters. Although infants carrying at least one E4 allele had higher LDL-C (as well as TC and triglyceride) levels than those carrying at least one E2 allele, these differences were not statistically significant. No significant differences in thyroid hormones were noted in E4 allele carriers in comparison with other patients. CONCLUSIONS: The observed lack of a significant correlation between thyroid hormones (except T3), apoE phenotypes and lipoprotein levels suggests that, early in infancy, other factors may play a more important role in determining lipoprotein levels. PMID- 10365573 TI - Effects of exercise for 1 month on serum lipids in adolescent females. AB - BACKGROUND: The present study was done to clarify the effects of 1 month of exercise on levels of total cholesterol (TC) and high-density lipoprotein cholesterol (HDLC) and on the ratio TC/HDLC and also to evaluate the relationship of body fat to amount of exercise and TC/HDLC ratio. METHODS: Twenty-seven female athletes (aged 15-18 years) were divided into two groups: the participant's group, in which players trained and attended a tournament, and the non participant's group, in which players did not attend the tournament. We assessed the amount of exercise, body composition, serum TC and HDLC and TC/HDLC on four occasions: before (T0), during (T1), 1 day after (T2) and a week after (T3) the experimental period. Levels of TC and HDLC adjusted for changes in plasma volume were compared for each occasion. Two multiple regression models for change in TC/HDLC from T0 to T2 and from T0 to T3 were employed. RESULTS: (i) The changing patterns in TC and HDLC throughout the program were different between the two groups; (ii) the decreased level of TC/HDLC after 1 month of exercise may easily revert to its original level; and (iii) the relevant factor for the decline in TC/HDLC was the amount of exercise, not body fat reduction. CONCLUSIONS: Exercise may be a more important factor for the improvement of TC/HDLC than concomitant body fat reduction and non-strenuous exercise may maintain a more stable and higher HDLC level than strenuous exercise. PMID- 10365574 TI - Significance of the atherosclerogenic index and body fat in children as markers for future, potential coronary heart disease. AB - BACKGROUND: The purpose of this study is to establish a simple marker in children for future, potential risks of coronary heart disease. METHODS: We measured serum total cholesterol (TC) and triglyceride (TG) by enzymatic methods, high-density lipoprotein cholesterol (HDLC) by the dextran sulfate-magnesium method and estimated body fat by the new impedance method in 1289 children (651 boys and 638 girls) in the fourth grade (9 or 10 years old) to obtain the atherosclerogenic index (AI). We also investigated the children's lifestyle. RESULTS: The probability of an AI score of 3 or more was significantly higher in children with an estimated 23-25% body fat than in those with body fat less than 17%. Moreover, the odds ratio increased along with an increase in the percentage of body fat. When body fat was estimated as being greater than 29%, the odds ratio was 11-fold higher than those with body fat less than 17%. When body fat was greater than 23%, the children's physical activity, as assessed by the questionnaire, was found to be poorer than those with lower body fat. Levels of TC and TG were significantly higher and that of HDLC was lower in those with less body fat. CONCLUSIONS: The AI is a useful indicator of obesity in children. The combination of AI and percentage body fat is a good indicator for evaluating children who would be at a greater risk of obesity, hyperlipidemia, unhealthy eating habits and inadequate physical activity. The hypothetical risk levels for future coronary heart disease are an AI score of > or = 3 and percentage body fat > or = 23% in Japanese children. PMID- 10365575 TI - Thyroxine inversely regulates serum intermediate density lipoprotein levels in children with congenital hypothyroidism. AB - BACKGROUND: Although there have been numerous studies on the effects of thyroid hormones on serum lipid profiles, the effects of thyroxine on intermediate how density lipoprotein (IDL) remain uncertain. In an attempt to clarify, this issue, under conditions with very little influence exerted by sex hormones on serum lipid profiles, we studied the relationship between serum thyroid hormone levels and the proportion of serum IDL fractions in children. METHODS: Nineteen children with congenital hypothyroidism and 13 children with non-thyroid diseases were enrolled in this study. Blood samples were taken to measure serum thyroid stimulating hormone, triiodothyronine, free thyroxine (FT4), total cholesterol, high density lipoprotein (HDL) cholesterol, triglyceride and apolipoprotein levels. Lipoprotein fractions, including very low density lipoprotein (VLDL), IDL, low density lipoprotein (LDL) and HDL, were determined by their electrophoretic mobility in a non-denaturing polyacrylamide gel. RESULTS: The proportion of IDL fractions showed a significant inverse correlation with serum FT4 levels and a significant correlation with serum total cholesterol and apolipoprotein B and C-II levels. Serum VLDL, LDL and HDL fractions did not correlate with serum thyroid hormone levels. CONCLUSION: From these results and other studies, we suggest that thyroxine promotes the conversion of IDL into LDL, possibly by its stimulatory effects on hepatic lipase activity. PMID- 10365576 TI - Efficacy of interferon therapy on serum fibronectin levels in children with chronic hepatitis B infection. AB - BACKGROUND: Fibronectin (FN) is a glycoprotein, the major sources of which are hepatocytes, Kupffer cells and endothelial cells. It has many biological functions including adhesion between cells, immunity, blood coagulation and platelet aggregation. Serum FN levels are generally decreased in pathological blood coagulation and inflammation. In the present study, we evaluated the serum levels of FN in patients with chronic hepatitis B virus (HBV) infection treated with interferon-alpha 2b. METHODS: We studied serum levels of FN in a prospective trial between October 1995 and May 1997. The study included 16 patients with chronic HBV infection before and after interferon therapy, in a period of 6 months, and 17 healthy controls. In total, we had 40 patients with chronic HBV infection. We studied these 16 patients (40%) who recovered with interferon therapy. We could not study the other 24 patients because we did not have enough of the reagents for studying FN. RESULTS: Chronic hepatitis B infection was diagnosed serologically and histopathologically. In mean age and sex, no statistically significant differences were found between patients and healthy subjects. The serum FN concentration before treatment with interferon therapy appeared significantly lower in HBV patients than in healthy control subjects (P = 0.026 using the Mann-Whitney confidence interval and test). After treatment with interferon, serum levels of FN were significantly higher than levels obtained before interferon therapy (P = 0.004 using the Wilcoxon Test). CONCLUSIONS: These results suggest that a decreased level of serum FN in patients with chronic hepatitis before interferon treatment is related to hepatic injury and inflammation. Because of inflammation, the serum FN level is decreased due to the consumption of FN. Increased levels of serum FN in patients having interferon therapy is important and is related to the effects of interferon including antiviral, antiproliferative, anti-inflammatory and immunoregulatory properties in patients with chronic HBV infection. A Japanese study showed a correlation between development of hepatic fibrosis and decrease of plasma FN concentration in adult patients with chronic liver disease. Therefore, the serum level of FN may be a useful marker of hepatic fibrosis in chronic liver disease and interferon may be an important drug for prevention of liver fibrosis. Fibronectin may be also a useful marker in predicting IFN response. PMID- 10365577 TI - Efficacy of antibiotics against influenza-like illness in an influenza epidemic. AB - AIM: To determine if an antibiotic reduces the incidence of complications associated with influenza-like illness during an influenza epidemic. METHODS: During the outbreak of influenza in Kobe in 1998, 85 patients suffering from an influenza-like illness were randomly assigned to two groups. Patients received placebo or sultamicillin orally for 4 days. The incidence of complications of influenza-like illness were compared and statistically assessed. RESULTS: There was no difference in the duration of fever or the incidence of acute otitis media. However, the incidence of pneumonia was significantly lower in the sultamicillin group than the placebo group (2.4 vs 16.3%, P < 0.05). CONCLUSION: Sultamicillin reduced the incidence of pneumonia associated with influenza-like illness during the influenza epidemic. This result suggests that antibiotics can reduce the rate of pneumonia associated with influenza. PMID- 10365578 TI - Incidence of Haemophilus influenzae in the throats of healthy infants with different feeding methods. AB - BACKGROUND: Haemophilus influenzae is the major cause of otitis media and lower respiratory tract infection in childhood. In the presence of human milk, which contains numerous host defense factors, Haemophilus influenzae may be inhibited in attaching to and colonizing pharyngeal cells. We investigated the incidence of H. influenzae in the throats of 162 healthy infants with different feeding methods: 70 breast-fed, 49 mixed-fed and 43 formula-fed infants. METHODS AND RESULTS: Haemophilus influenzae was identified using standard microbiological procedures and the API NH system. The incidence of H. influenzae in breast-fed infants, mixed-fed infants and formula-fed infants was 0, 0 and 7.0% respectively. CONCLUSION: The results suggest that the colonization of H. influenzae in the throat was inhibited by the presence of breast milk. PMID- 10365579 TI - Transient galactosemia detected by neonatal mass screening. AB - BACKGROUND: The Paigen method has detected not only persistently galactosemic patients, but also many children with transient galactosemia during the neonatal period. The diagnosis and clinical course of 389 patients with transient galactosemia detected by neonatal mass-screening from 1986 to 1996 in the Hiroshima prefecture were evaluated. METHODS: Enzyme assays for galactose metabolism, measurement of blood galactose levels, erythrocyte galactose-1 phosphate levels, serum total bile acid (TBA) levels and liver function tests were performed at the first visit by patients to our hospital. Liver function and the mental and physical development of patients were evaluated during the follow up period (approximately 1 year). RESULTS: The diagnoses were classified as follows: 253 patients with unknown cause, 128 heterozygotes and two homozygotes for galactose enzyme deficiency (galactose-1-phosphate uridyltransferase, galactokinase, UDP-galactose 4-epimerase) and six heterozygotes for Duarte variant. Twelve patients showed high serum levels of TBA (> 80 mumol/L), which suggests the presence of portal-systemic shunts during the neonatal period causing galactosemia. Most patients showed normal mental and physical development during infancy. However, of 25 patients with mild to moderate abnormal liver function tests of unknown etiology after the neonatal period, five showed poor weight gain coincident with liver dysfunction. In almost all patients, levels of transferase decreased to the normal range by 1 year of age. CONCLUSION: We found that the prognosis of transient galactosemia was almost always favorable. However, patients should be followed for at least 1 year, because late liver dysfunction, which might cause poor weight gain, occurred in 6% of our patients. PMID- 10365580 TI - Contingent negative variation in children with anorexia nervosa. AB - BACKGROUND: Central catecholamines, particularly dopaminergic and noradrenergic systems, have affected the appetitive behavior in patients with anorexia nervosa (AN). The purpose of this study is to distinguish the characteristics of contingent negative variation (CNV) and postimperative negative variation (PINV), which may reflect the level of catecholamine in children with AN. METHODS: Eight children with AN aged 10 to 15 years and 23 age-matched healthy children were recruited. Contingent negative variation was recorded from the frontal midline (Fz), central midline (Cz) and parietal midline (Pz) referenced to linked earlobes during 30 trials consisting of a warning stimulus and an imperative stimulus with an interstimulus interval of 2 s and an intertrial interval of 10 s. The imperative stimulus of each trial required a button press. RESULTS: Children with AN had a diminished amplitude of the CNV. They had a significantly more attenuated early CNV and late CNV amplitude at Cz than normal children. No significant differences were observed between AN children and normal children in the amplitude of PINV at all three electrode sites. No difference could be found between the two groups in the frequencies of normal and abnormal duration of PINV. CONCLUSION: These findings suggest that early CNV may be diminished by norepinephrine deficiency and late CNV may be attenuated by dopaminergic deficiency in children with AN. Reduced CNV may represent impaired cognitive processes which reflect impaired appetitive behavior in AN children. PMID- 10365581 TI - Two-dimensional autoregressive analysis of carotid artery blood flow waveform in children with isolated atrial septal defect. AB - BACKGROUND: The aim of the present study was to analyze the carotid artery blood flow waveform, using a two-dimensional autoregressive modeling approach and component analysis, and to determine the relation between cardiac contractility, peripheral and cerebral circulation and characteristic values of component activities of carotid artery blood flow waveform in patients with atrial septal defect (ASD), with or without congestive heart failure. METHODS AND RESULTS: We analyzed the carotid artery blood flow waveform of nine patients with ASD and 35 normal controls using a two-dimensional autoregressive modeling approach. The component of impulse response was divided into six groups according to the damping frequency: (i) group I, 0 Hz; (ii) group II, 1-5 Hz; (iii) group III, 5-8 Hz; (iv) group IV, 8-13 Hz; (v) group V, 13-17 Hz and (vi) group VI, > 17 Hz. The decrease of impulse response power-density in patients of groups I, II, III and IV and the prolongation of damping time for patients in groups I and II were particularly noticeable in two ASD patients, whose pulmonary to systemic blood flow ratio was more than 2.7 and whose left ventricular stroke volume was less than 33.1 mL/m2. The power-density of groups I and II varied with cardiac contractility and the power-density of groups III and IV varied with cerebral circulation. In contrast, the damping time of groups I and II changed with the reflection velocity from the position of arterial reflection against blood flow from left ventricle. CONCLUSIONS: These results may be influenced by the decrease in left ventricular stroke volume and velocity in arterial reflection. PMID- 10365582 TI - Bartter's syndrome in Arabic children: review of 13 cases. AB - BACKGROUND: Bartter's syndrome (BS) is an inherited disease of renal potassium wasting characterized by hypokalemic alkalosis, normal blood pressure, vascular insensitivity to pressor agents and elevated plasma concentrations of renin and aldosterone. It is caused by generalized hyperplasia of the juxtaglomerular apparatus at the site of renin production caused by mutations in the Na-K-2Cl cotransporter gene, NKCC2. The objective of our study is to establish the prevalence and incidence of BS in Kuwait and to assess treatment modalities for it. METHODS AND RESULTS: Bartter's syndrome was diagnosed in 13 Kuwaiti children over a 14 year period (1981-1995) with the estimated incidence of 1.7/100,000 live births. The mean age at diagnosis was 9.3 months (range 2-32 months). There were five males and eight females (ratio 1:1.6). The mean duration of follow up was 5.6 years (1-14 years). Both consanguinity and familial history among our patients were high (69 and 54%, respectively). All patients had hypokalemia, hypochloremia with metabolic alkalosis, hyperreninemia and were normotensive. Clinical presentation was essentially similar to that in other series. Eleven patients (85%) had growth failure, two had nephrocalcinosis (15%) and one had renal failure. All patients were treated with supplemental potassium, an aldosterone antagonist (spironolactone) and a prostaglandin synthetase inhibitor (indomethacin or aspirin) sequentially. Significant catch-up of growth (four patients) and increases in serum potassium (eight patients) were recorded after administration of indomethacin therapy. One patient died of severe pneumonia with respiratory failure from hypokalemic myopathy. Clinical presentation, inheritance, complications and therapy of BS are briefly discussed. CONCLUSION: Bartter's syndrome is a rare disease, but should be considered in the differential diagnosis of other disorders with growth failure and/or hypokalemia. Early diagnosis, close follow up and compliance with treatment may lead to appropriate growth and development. PMID- 10365583 TI - Rebound cerebrospinal fluid pleocytosis with elevated cytokine levels during recombinant human granulocyte colony-stimulating factor therapy for purulent meningitis. PMID- 10365584 TI - Neonatal meningitis due to a vertical transmission of Pasteurella multocida. PMID- 10365585 TI - Natural emergence of an anti-hepatitis B s escape mutant in a young female hepatitis B virus carrier. PMID- 10365586 TI - Atypical Epstein-Barr virus infection associated with Gianotti-Crosti syndrome and Bell's palsy. PMID- 10365588 TI - Gastroesophageal reflux associated with dystonic movements: Sandifer's syndrome. PMID- 10365587 TI - Rupture of coronary aneurysm in Kawasaki disease. PMID- 10365589 TI - Kikuchi's disease with leukocytoclastic vasculitis in a 10-year-old girl. PMID- 10365590 TI - A case of congenital nasal pyriform aperture stenosis. PMID- 10365592 TI - [Antiarrhythmic agents in the prevention of sudden cardiac death]. AB - Sudden cardiac death due to ventricular arrhythmias remains a significant problem. In most studies about 50% of all death related to coronary artery disease and heart failure are sudden and unexpected and are caused by acute fatal ventricular tachycardia and fibrillation. Most of the patients suffering sudden cardiac death have some kind of structural heart disease but 80% of SCD events are associated with coronary artery disease, 10-15% with dilated and hypertrophic cardiomyopathy, and only small fraction with the less common disorders as valvular heart disease, ventricular dysplasia and cardiac involvement in sarcoidosis or amyloidosis. In some patients the anomaly responsible for sudden cardiac death is not structural but mainly electrical as in patients with the long QT syndrome, WPW syndrome or in patients with a proarrhythmic effect from antiarrhythmic drugs. In this review, data from clinical trials and other studies on on antiarrhythmic therapies have been evaluated in order to determine effective strategies for the prevention sudden cardiac death in high risk patients. Taken together with the mortality data routine prophylactic use of class I antiarrhythmic drugs in the patients survivors of acute myocardial infarction and patients with heart failure is associated with increased risk of death. Conversely beta-blockers are associated with significant reduction in nonfatal cardiac arrest in the short term trials and sudden cardiac death in long term trials. These benefits are likely due to relief ischemia, reduction of heart rate and maintenance favourable autonomic nervous system balance. Overall trial data on amiodarone suggests that this agent is effective in reducing the risk of death in survivors of cardiac arrest, post infarction patients, and patients with heart failure but the routine prophylactic use of amiodarone remains of uncertain efficacy. The physician who considers the use of antiarrhythmic medications in patients with ventricular arrhythmias must be aware of which arrhythmias are malignant or potentially malignant and which are benign and the decision to initiate antiarrhythmic therapy should be based on consideration of the patients absolute mortality risk. PMID- 10365591 TI - Increased circulating granulocyte colony-stimulating factor in acute Kawasaki disease. PMID- 10365593 TI - [The evaluation of the coronary artery sufficiency in myocardial infarction patients after cardiac surgery and preventive therapy]. AB - During 5 years period (1989-1993) the authors investigated a group of 40 patients with coronary artery disease after myocardial infarction treated in the Silesian Center of Cardiology in Katowice. 20 patients were operated on (CABG), 20 were medically-treated. It was evaluated the history, physical status, stress-test, echocardiography and 24-hours ECG. Stress test was estimated according to Mark's test. In the echocardiographic examination it was observed wall motion score index (WMSI) and the left ventricular abnormal contraction area (AA). In the operated group it was noticed higher physical ability and no influence of CABG on left ventricular contractability. PMID- 10365594 TI - [Studies of lymphocyte membrane transport of sodium in patients with essential hypertension]. AB - The aim of this study was to investigate abnormalities in lymphocyte membrane sodium fluxes in patients with essential hypertension with and without familial history of hypertension and the influence of selected hypotensive drugs on these fluxes. 121 patients (pts) with positive family histories of primary hypertension (PFH) and 73 pts with negative family histories of primary hypertension (NFH) were examined. The total sodium efflux rate constant (wswc), ouabaine-sensitive (wswou) and furosemide-sensitive (wswf) were measured by the method of Heagerty et al. To examine the influence of selected hypotensive drugs on sodium fluxes wswc, wswou and wswf were measured before and after 7 days of treatment with hydrochlorothiazide (H) or propranolol (P). Wswou was decreased in 61% pts with PFH and in 19% pts with NFH, wswf was decreased in 38% pts with PFH and in 22% pts with NFH. Both, wswou and wswf, were decreased in 49% pts with PFH and only in 2.7% pts with NFH. Wswou and wswf rose significantly after 7 days of treatment with H or P only in pts with PFH and in pts with decreased wswou and wswf before treatment. These data suggest that abnormal lymphocytes membrane sodium transport often occurs in pts with PFH and has familial component. Changes in transport systems observed after 7 days treatment with H or P may contribute, at least in part, to its antihypertensive action in familial hypertension. PMID- 10365595 TI - [Changes of lipid profile in elderly people in light of longitudinal studies]. AB - The purpose of the study was a retrospective assessment of the health state, dietary habits and nutritional status of elderly men and women. Particular attention was paid to changes in lipid profile of the elderly over 5 years of life in a big urban agglomeration. The first study was carried out in 1992 in a population sample of 154 people aged 70 years (73 men and 84 women). After five years the study was repeated in the same sample. In 1997 for the studies 103 persons came (43 men, 60 women). In the 5-year period 13 persons died. The study showed that over that time period the total cholesterol level, HDL and LDL cholesterol decreased. In the same time period there was a decrease in the consumption of fats and cholesterol in the diet. The fall of HDL cholesterol level caused an increase of the atherogenicity index in these people increasing thus the likelihood of ischaemic heart disease development in this group. PMID- 10365596 TI - [The beneficial effects of growth hormone replacement therapy on elderly men]. AB - The relationship of growth hormone (GH) to the ageing process is currently subject of considerably interest. The study was designed to investigate the effects of replacement therapy with growth hormone on quality of life, serum lipids and body composition (fat free mass and fat mass) in elderly men. MATERIAL: 18 healthy men 60.0 +/- 2.4 (x +/- SEM) years of age. Their body weight was 78.6 +/- 4.6 kg and body mass index (BMI) was 26.5 +/- 1.4 kg/m2. Diagnosis of GH deficiency was based on serum insulin-like growth factor-1 (IGF-1) levels below 200 micrograms/L (138.1 +/- 9.2), abolished GH nocturnal surge and diminished glucagon-stimulated GH secretion compared to reference group of young men (16.2 +/- 1.8 to 30.6 +/- 4.7 micrograms/L/hour; p < 0.02 and 10.8 +/- 1.0 to 44.1 +/- 15.3 micrograms/L/hour; p < 0.02, respectively). Reference group comprised nine men 27.5 +/- 1.3 years of age with body weight 76.3 +/- 2.2 kg and BMI 23.1 +/- 0.6 kg/m2. The subjects received recombinant, human GH daily subcutaneously during 12 months in dose adjusted to maintain optimal (280-350 micrograms/L) serum IGF-1 level. The initial dose was 0.125 IU/kg b.w./week. Before, and after 6 and 12 months of therapy clinical and laboratory exams, including serum GH, IGF-1 and lipids levels, and body composition using two methods were obtained. Quality of life was assessed by modified Beck's questionnaire. 12-months replacement therapy with growth hormone in elderly men improved mental status, increased serum IGF-1 level to the young normal men values, from 138.1 +/- 9.2 to 279.4 +/- 26.3 micrograms/L, p < 0.001, reduced serum LDL-cholesterol from 3.67 +/- 0.12 to 3.10 +/- 0.21 mmol/L, p < 0.04 and increased serum HDL and HDL2 levels from 1.20 +/- 0.05 to 1.41 +/- 0.08 mmol/L, p < 0.002 and from 0.19 +/- 0.03 to 0.34 +/- 0.06 mmol/L, p < 0.005, respectively, reduced fat mass (12.8%, p < 003), particularly localised in trunk (14.7%, p < 0.03), and increased fat free mass (2.9%, p < 0.03). GH-replacement therapy in elderly men has beneficial effects on quality of life, and may counteract ageing and atherosclerosis progression by serum lipids and body composition improvement. PMID- 10365597 TI - [The influence of ozone therapy on endothelial damage markers in patients with atherosclerosis of lower extremities]. AB - The aim of study was to evaluate the influence of the treatment with oxygen-ozone mixture on the blood plasma antigen concentration of tissue plasminogen activator (t-PA) and von Willebrand factor (vWF) in patients suffering from atherosclerotic disease of lower extremities. The study was performed in the group of 28 (M/F 22/6) patients means aged 64.1 years with atherosclerotic diseases of lower extremities, in whom 2 weeks therapy with oxygen-ozone mixture was used. The control group consisted of 30 healthy volunteers in mean age 51.0 years. In the blood plasma obtained from the patients before and after treatment with oxygen ozone mixture and from the control group determinations of t-PA and vWF antigen using ELISA were done. Both parameters were significantly increased in the patients before the treatment in comparison to the healthy controls. The treatment with oxygen-ozone therapy caused in patients slight statistically not significant raise of t-PA and vWF antigen showing the endothelial stimulation but not the destruction of vascular endothelium. PMID- 10365598 TI - [Magnesium and zinc levels in blood serum and cerebrospinal fluid in children with febrile convulsions]. AB - Magnesium and zinc levels in blood serum and cerebrospinal fluid were estimated in 18 children aged 8 months-5 years with febrile convulsions. Control group consisted of 15 apparently healthy children in the same age. On the ground of ethical reasons no control values for cerebrospinal fluid were evaluated. The mean serum concentration of magnesium and zinc was significantly lower in the sick children (p < 0.001 and p < 0.05 resp.). The magnesium levels in cerebrospinal fluid were evidently lower than those quoted in the literature and the mean concentration of zinc was higher in comparison with literature data. The presumable pathogenetic role of disturbed magnesium and zinc metabolism in febrile convulsions is discussed. PMID- 10365599 TI - [Chylopericardium as a complication of mitral valve replacement]. AB - We demonstrate the case of 47 year old woman with chylopericardium after mitral valve replacement. By reason of heart tamponade she underwent surgical treatment resulted in complete relief of the pericardial effusion. PMID- 10365600 TI - [The dural arteriovenous fistula treatment by embolization]. AB - The transverse sinus is one of the most common locations for dural fistulas. A 43 year-old man with pulsatile tinnitus had arteriography that showed a left transverse dural arteriovenous fistula. We report on the treatment of the fistula with embolization of the supplying arteries. Primary transarterial embolization failed to completely obliterate the fistula and the recurrent symptoms were observed. Subsequent embolization was tried by using PVA (polyvinyl alcohol), spongostan and mechanical detachable coils and resulted in the improvement of the patient's tinnitus. The additional arteriographical finding was microangioma located on the branch supplying the fistula arising from the left vertebral artery. PMID- 10365601 TI - [Mechanical extracorporeal circulation. Artificial heart. Percutaneous cardiopulmonary circulation. Part II]. AB - Problems of extrcorporeal circulation are discussed in relation to newer methods and techniques applied in emergency. PMID- 10365602 TI - [Trends in pharmacological treatment of congestive heart failure]. AB - Congestive heart failure (CHF) is growing epidemiologic and clinical problem, and is the only common cardiovascular condition that is increasing in incidence, prevalence and mortality. During last years numerous clinical trial have been conduced evaluating the effect of various treatment procedures on clinical endpoints in patients with CHF. The major risk factor for CHF are hipertension and atherosclerotic vascular diseases, and now it is clear that aggressive treatment of hypertension and hyperlipidemia can be effective in preventing CHF. Treatment strategies for CHF are aimed at preventing and delaying progression of the disease and improving survival. In the treatment of CHF diuretics are at present the first drugs line for patients with fluid retention and are necessary to relieve symptoms but cannot halt progression or improve the prognosis of CHF. Angiotensin-converting enzyme inhibitors (ACE inhibitors) therapy has been shown to decrease mortality and progression of CHF and should be used early in patients with left ventricular dysfunction whether they have symptomatic or asymptomatic CHF. Digoxin therapy is associated with decrease in the risk of worsening CHF irrespective of rhythm, systolic function, severity of CHF or therapy with ACE inhibitors. In patients with symptomatic CHF due to systolic dysfunction the addition of diuretics and digoxin appears to reducing worsening CHF without improving survival. Other than digoxin oral inotropic agents (amrinone, pimobendan, vesnarinone, ibopamine) increase mortality in patients with CHF and have not improved symptom status and other clinical endpoints during long-term therapy. Hydralazine and isosorbide dinitrate administrated in combination are less effective alternative to ACE inhibitors. Beta-blockers and particular carvedilol may prolong survival and decrease worsening CHF when used in combination with digoxine, diuretics and ACE inhibitors. Beta-blockers therapy improve hemodynamics, LVEF and functional status patients with CHF and the ideal candidate for this therapy is stable patients with NYHA II-III CHF due to nonischemic cause. Calcium antagonists do not appear to be useful in patients with CHF, although amlodipine and mibefradil appears to be safe for treatment of angina or hypertension in this group. On the basis of current data, antiarrhythmic agents should not be given to patients with CHF free from arrhythmia but those with sustained ventricular tachycardia or ventricular fibrillation amiodaron appears to be safe. PMID- 10365603 TI - [Thyroid hormones and the heart]. AB - The thyroid hormones exert effects on the heart and the peripheral circulation playing an important role in the regulation of the function of the sinoatrial node, the systolic and diastolic function of the myocardium and the peripheral resistance. Their act directly by influence on protein synthesis, the properties of cell membranes and indirectly by interactions with autonomic nervous system causing increase in cardiac output and decrease systemic vascular resistance. Applying thyroid hormone therapy to the treatment of various forms of heart disease is connected with the development of the knowledge about their mechanism of action. PMID- 10365604 TI - [Magnesium: its significance in cardiology]. AB - The significance of magnesium in the etiology and treatment of diseases of the circulatory system has been investigated for several decades. Experimental, clinical and epidemiological data concerning this problem are abundant, however their findings do not let--at least so far--to draw unequivocal conclusions as to the significance of this element in the treatment of cardiovascular diseases. Mg++ is known to be responsible for the function of about 300 enzymes, and its deficiency results in necrosis of the cells due to depletion of energetic stores of phosphates, leads to disturbances in fat metabolism, increased aggregation and shortened survival time of platelets, increased level of factor III, disturbed synthesis of proteins (mitral valve leaflet prolapse) as well as of prostanoid function. Despite the fact that serum level of Mg++ differs from its concentration in tissues, majority of clinical studies relies on serum levels of the element. The effects of changes in serum levels of Mg++ resulting from various pathophysiological processes should be differentiated from the effects of magnesium supplementation or from administration of the element in excess for therapeutic reasons when the levels in the organism have been considered normal. PMID- 10365605 TI - [The influence of angiotensin converting enzyme inhibitors on the central nervous system]. AB - Angiotensin converting enzyme inhibitors (ACEI) constitute 23% of all antihypertensive drugs world-wide (WHL Newsletter, October 1997). Although lipophilic ACEIs like captopril, trandolapril or perindopril may decrease blood pressure partly via central nervous system (CNS) they also are capable of evoking a range of psychotropic effects. Basing on our recent experiments and the literature data we discussed possible effects of ACEIs on mood and cognition. PMID- 10365606 TI - [The assessment of the functional state of elderly people by family physician with the help of EASY-Care questionnaire]. AB - The multiplicity and complexity of diagnostic and therapeutical problems affecting elderly patients requires a special approach which has come to be known as the comprehensive geriatric assessment. This approach demands the interdisciplinary cooperation of several practitioners in the field of geriatric care, led by the family doctor. An essential prerequisite for their cooperation to be successful is that there should be some commonly acknowledged starting point in the form of a standardised assessment of the functional state of elderly patients, irrespective of the the number and type of chronic conditions from which they may be suffering. The aim of this paper is to put forward a short and straightforward instrument for assessment of the functional state of elderly people in the form of the EASY-Care questionnaire for the use of family doctors. EASY-Care is a system "picture" of the needs of elderly people. It focuses more on the quality of life of the elderly person rather than on his/her illnesses and takes into consideration the role played by family carers. EASY's main aim is to provide family doctors and other primary care practitioners with information to help them to improve the services that they provide for the elderly. Moreover, the data gained from the use of EASY may help to serve epidemiological purposes, by providing measures of the social and medical needs of the elderly together with information about the degree to which these are being met. Consequently, it also provides a rationale for the provision of funding, directing research and developing a policy for the care of the elderly. The EASY-Care questionnaire has been developed in order to promote a common approach in Europe for the assessment of the elderly. PMID- 10365607 TI - [Patient satisfaction from the contact with the physician]. AB - The satisfaction of a patient at the contact with a doctor results from realisation of patient's emotional and medical needs by the doctor. Lack of satisfaction reduces patient's ability to acquire and memorize information, causes unwillingness to follow doctor's instructions, eliminates the need of prophylactic examinations and decreases the level of rational response to disease symptoms. It is an emotional barrier which often makes it impossible to archive therapeutic aim. PMID- 10365608 TI - [Postoperative emesis. Reasoning the strategy to ration the expense; comment]. PMID- 10365609 TI - [Ondansetron in the prophylaxis of postoperative nausea and vomiting in ambulatory cataract surgery]. AB - INTRODUCTION AND OBJECTIVE: Postoperative nausea and vomiting (PONV) are potentially serious complications of ophthalmic surgery. We assess the efficacy of ondansetron for antiemetic prophylaxis in outpatient unilateral cataract surgery under retrobulbar blockade. PATIENTS AND METHODS: Cohort study of patients undergoing unilateral cataract surgery between January 1996 and March 1997. The main predictive variable was intravenous administration of 4 mg of ondansetron 30 min before surgery and the main effect variable was the presence of PONV during the first 24 h after surgery. The incidence of PONV was calculated and an analysis of statistical significance was performed using a Mantel-Haenszel chi 2 test, describing the magnitude of association between relative risk and the corresponding confidence interval (95% CI). RESULTS: One hundred sixty-two patients were enrolled. Eighty-two patients received ondansetron and 80 did not. The two groups were similar with respect to control variables. PONV occurred in 23 patients (14.2%): in 16 (20%) who did not receive ondansetron and in 7 (8.2%) who did (p < 0.05). The relative risk of patients who received ondansetron was 0.42 (95% CI: 0.19-0.98) in comparison with those who did not. CONCLUSION: We found a high incidence of PONV, although the administration of ondansetron reduced PONV significantly. PMID- 10365610 TI - [Conditions of intubation and neuromuscular block induced by mivacurium: comparison with succinylcholine]. AB - OBJECTIVE: To compare the clinical conditions for intubation and neuromuscular parameters after a high dose of mivacurium (0.25 mg/kg; 3 x SD95) administered in 30 s to those obtained after use of the usual dose of succinylcholine (1 mg/kg). PATIENTS AND METHODS: Eighty-two patients, 37 in the succinylcholine group and 45 in the mivacurium group, were studied. Intubating conditions were assessed on a scale of 3 to 12 points analyzing ease of laryngoscopy, relaxation of vocal cords and presence of cough 60 seconds after administration of the drug. Neuromuscular parameters were acceleration of the thumb induced by supramaximal train-of-four stimulation of the cubital nerve. Heart rate and non-invasive mean blood pressure were recorded throughout surgery. Cutaneous flush was looked for after administration of the relaxant. RESULTS: Time to onset of effect (159 s versus 82 s) and times to recovery after mivacurium were significantly longer than with succinylcholine. Mivacurium afforded excellent/good conditions for intubation in 95.6% of cases, with a neuromuscular blockade at intubation of 52.68%. No significant hemodynamic changes or side effects were observed. CONCLUSIONS: Given the moderate conditions of intubation achieved at 60 s, mivacurium can not be recommended as a relaxant in situations that require a rapid induction sequence. In elective surgery, 0.25 mg/kg of mivacurium can, however, be considered an alternative to succinylcholine. PMID- 10365611 TI - [Intravenous anesthesia with propofol in intracranial surgery]. AB - OBJECTIVES: To analyze the repercussions of intravenous anesthesia with propofol as the single hypnotic drug on intracranial pressure (ICP) and cerebral perfusion pressure (CPP), and also to study the time until recovery from anesthesia and to tracheal extubation as well as intraoperative hemodynamic changes in patients undergoing surgery to remove a supratentorial brain tumor. PATIENTS AND METHODS: Twenty-three ASA I/II patients scheduled for exeresis of a supratentorial brain tumor were studied. A fiberoptic sensor placed in direct contact with the dura mater was used to measure ICP. Anesthetic induction was achieved with propofol (2 mg/kg). Propofol (12 and 9 mg/kg/h for 10 min and 6 mg/kg/h throughout the rest of the operation) was used for maintenance. Mean arterial pressure (MAP), heart rate (HR), ICP and CPP were recorded at baseline and 1, 2, 3 and 4 min after induction, during laryngoscopy and tracheal intubation; 1, 3, 5, 10, 15 and 20 min after tracheal intubation (L + 1, L + 3, L + 5, L + 10, L + 15, L + 20), upon placement of a craniostat; upon skin incision; upon withdrawal of propofol perfusion; and during extubation. The following variables were recorded after awakening: time until eye opening after receiving a verbal command, time until extubation and time until orientation. Analysis of variance for repeated measures (ANOVA) was performed on the results. RESULTS: MAP decreased significantly from baseline at the following times: during the post-induction period, upon placement of the craniostat, upon skin incision and when the propofol infusion was switched off. HR increased significantly during laryngoscopy and at the following moments: intubation, post intubation (L + 1, L + 3, L + 5), craniostat placement, and extubation. ICP was lower throughout the surgical period except during laryngoscopy, when this variable increased significantly. CPP decreased significantly after induction and returned to baseline after intubation. CPP was significantly higher after surgery. Recovery times after weaning from propofol infusion until eye opening in response to an order and until orientation were 13 +/- 3 and 22 +/- 4 min, respectively. The mean interval between withdrawal of propofol until extubation was 18 min. CONCLUSIONS: Intravenous anesthesia with propofol in intracranial surgery (supratentorial tumors) affords hemodynamic stability and lowers ICP except during laryngoscopy. Early recovery from anesthesia allows for neurological assessment and vigilance during the immediate postoperative period. PMID- 10365612 TI - [Comparative study of 3 techniques for total intravenous anesthesia: midazolam ketamine, propofol-ketamine, and propofol-fentanyl]. AB - OBJECTIVE: To compare the characteristics of induction, maintenance and awakening for three techniques of combined total intravenous anesthesia (TIVA): propofol ketamine, midazolam-ketamine and propofol-fentanyl. PATIENTS AND METHODS: Sixty patients were randomly assigned to three TIVA groups. Group 1 (n = 20) received midazolam, ketamine and vecuronium. Group 2 (n = 20) received propofol, ketamine and vecuronium. Group 3 (n = 20) received propofol, fentanyl and vecuronium. The variables compared were hemodynamic changes during induction and maintenance and upon awakening; time until awakening; and the incidence of postanesthetic complications. We also assessed whether propofol was better than midazolam at preventing the psychomimetic effects of ketamine. RESULTS: The demographic characteristics of the three groups were similar. Hemodynamic variables were most stable in group 2. Perfusion of midazolam-ketamine was accompanied by a significantly higher number of hypertensive peaks. Time to awakening was significantly shorter in Group I (11.8 +/- 5 min) than in group 2 (20.2 +/- 12.5 min); in group 2 time to awakening was 16.6 +/- 5.6 min. Eight patients in group 1, 5 in group 2 and 1 in group 3 reported having bad dreams, the difference between groups 1 and 3 reaching statistical significance. No patient experienced hallucinations and all reported satisfaction with the anesthetic technique used. CONCLUSIONS: TIVA with ketamine and propofol is comparable to the most commonly used combination of propofol and fentanyl and may be an appropriate choice when hemodynamic stability is of great importance; withdrawal 15 min before ending surgery prevents prolonged awakening. Perfusion of midazolam-ketamine is not recommendable for scheduled surgery because it induces too many hypertensive peaks. Although neither midazolam nor propofol completely prevents the psychomimetic effects of ketamine, such effects are not so severe that patients reject the anesthetic technique used. PMID- 10365613 TI - [Intraoperative transesophageal echocardiography]. AB - Transesophageal echocardiography is being used increasingly by anesthesiologists for monitoring and diagnosis. Real-time imaging provides valuable information about anatomy, preloading and cardiac contractility. Its use is mandatory in valve repair surgery and it has been shown to detect cardiac ischemia before any other monitoring tool. Programs to teach transesophageal echocardiography to anesthesiologists should be implemented; the availability of backup support staff from the echocardiography unit is of great value. PMID- 10365614 TI - [Intraoperative diagnosis of a mediastinal paraganglioma: Anesthetic management]. AB - A 45-year-old woman with a posterior mediastinal tumor underwent right thoracotomy for resection, developing hypertension and difficult-to-control tachycardia while the tumor was being manipulated. A catecholamine-secreting tumor was suspected, the operation halted, and the patient prepared for surgery at a later time. The tumor was a mediastinal paraganglioma and the final outcome was satisfactory. Risk related to anesthesia is high in such patients, with perioperative mortality ranging from 40 to 85%. Correct diagnosis and appropriate preoperative drug preparation with adrenergic receptor blockers appreciably decreases morbidity and mortality related to surgery. We discuss the effect of labetalol on such tumors and describe our observations. PMID- 10365615 TI - [High-frequency jet ventilation with a proximal injector in a case of surgery of the tracheal carina]. AB - A 44-year-old man underwent surgery requiring medial sternotomy of resection of a tracheal carcina tumor. High frequency jet ventilation was used during tumor resection and reconstruction of the carina in order to shorten the time of surgery and to provide a nearly immobile and unobstructed surgical field. The only noteworthy complication was a tendency to respiratory acidosis that resolved without sequelae in the operating room once the trachea was closed. The anesthetic challenges presented by the various techniques used throughout history for such an interesting and complex type of surgery are briefly summarized. We also discuss the role of high frequency jet ventilation on surgery involving the major airways. PMID- 10365616 TI - [Renal cryopreservation in 2 renal transplant patients undergoing aortic surgery]. AB - Acute kidney failure is one of the most severe complications of surgery on the aorta. We describe two kidney-transplanted patients diagnosed of aneurysm of the abdominal aorta who underwent resection of the aneurysm and placement of an aorto aortic graft. Cryopreservation of the renal implant was used with good results. Prevention of kidney damage also requires maintenance of adequate intravascular volume and the use of drugs (dopamine, mannitol and furosemide) to increase renal blood flow and urinary output. PMID- 10365617 TI - [Use of labetalol in the anesthetic management of pheochromocytoma]. PMID- 10365618 TI - [Subcutaneous palpebral emphysema after laparoscopic herniorrhaphy]. PMID- 10365619 TI - [Importance of capnography in the early clinical diagnosis of malignant hyperthermia syndrome]. PMID- 10365620 TI - [Formation of a curl in a central venous catheter after catheterization of the basilic vein]. PMID- 10365621 TI - [Use of rocuronium in Duchenne's disease]. PMID- 10365622 TI - Prevalence, comorbidity, risk factors and service utilisation of disruptive behaviour disorders in a community sample of children in Valencia (Spain). AB - BACKGROUND: The importance of cultural and adverse family-environment variables as risk factors for Disruptive Behaviour Disorder has been repeatedly shown, and hence a variation in rates and risk factors between cultures could be expected. Lower rates should be found in countries with strong and stable family ties, such as Spain. OBJECTIVE: Prevalence rate, severity and comorbidity of Disruptive Behaviour Disorder, as well as risk factors and help-seeking behaviour relating to this disorder, were studied in a general population random sample of 387 10 year-old children living in Valencia (Spain). METHODS: DSM-III-R diagnosis was established by means of the KIDDY-SADS (Kiddy Schedule for affective diseases and Schizophrenia (epidemiological version)) interview and severity of the disorder was evaluated with the General Assessment Functioning (GAF) Scale. Other variables measured were: sex, number of siblings, parental occupation, single parent home, school failure, socioeconomic level, chronic somatic ailments and use of mental health services. RESULTS: Prevalence and severity parameters were low (for GAF70, prevalence = 11.1; for GAF60, prevalence = 4.9), albeit falling within the range reported in other countries. Morbidity profile and use of services did not substantially depart from the findings reported in other cultures. Different risk factors were associated with Attention Deficit Hyperactivity Disorder, Conduct Disorder and Oppositional Defiant Disorder, thus confirming the validity of considering them as separate dimensions. CONCLUSIONS: The findings did not support the hypothesis of lower rates and different risk factors and morbidity patterns in the Spanish sample studied. PMID- 10365623 TI - Risk factors for conduct disorder among Navajo Indian men and women. AB - OBJECTIVES: To describe the risk factors for conduct disorder before age 15 among Navajo Indians. METHODS: The study was based on a survey of a stratified random sample of adult Navajo Indians between the ages of 21 and 65 living on and adjacent to two different areas of the Navajo Reservation. There were 531 male and 203 female respondents. The average age (SD) of the men was 38.7 (10.5) years and of the women 35.5 (9.0) years. Conduct disorder was diagnosed retrospectively using the Diagnostic Interview Schedule first developed for the Epidemiological Catchment Area study. The responses were combined into a continuous scale. RESULTS: Significant risk factors for increased scores on the conduct disorder scale were: histories of physical and sexual abuse in childhood; abusive maternal drinking; a small number of households per camp; younger age; and being male rather than female. Measures of social status and religion in which subjects were raised were not significant. CONCLUSIONS: Many of the risk factors that are associated with conduct disorder in other populations are also risk factors in the Navajo population. There is suggestive evidence that some of these risk factors have become more common since World War II, raising the possibility that conduct disorder has become more prevalent, as is thought to be the case nationwide. PMID- 10365624 TI - Issues in the identification of comorbidity of mental retardation and psychopathology in a multicultural context. AB - Identification of the comorbidity of mental retardation and psychopathology in a multicultural setting raises manifold difficulties. The present study explored the sampling and identification issues implicated in estimating the prevalence of this dual diagnosis in a South African clinic sample. The relations between the prevalence of dual diagnosis and socioeconomic status, gender, and severity level of retardation were investigated. The detection rate of 4.36% was significantly lower than that of other studies. Prevalence was found to be greater in areas of high socioeconomic status, among males, and among less severely retarded individuals. Implications of these findings for cross-cultural studies and for allocation of service resources for patients with dual diagnosis are considered. PMID- 10365625 TI - Psychological distress and psychiatric disorders in primary health care patients in East and West Germany 1 year after the fall of the Berlin Wall. AB - BACKGROUND: The reunification of Germany confronted citizens in East and West Germany with many changes in their lives. These changes may be considered as critical life events. Especially for those in East Germany, life circumstances drastically changed, and individuals were increasingly required to adopt and develop coping capabilities. In addition to new opportunities and freedom, there was threatening uncertainty about the future. Theories of life events and stress postulate that threat events have an impact on human well-being. It was expected that there would be an increased rate of psychiatric morbidity after unification, especially in the eastern part of Germany. METHOD: An international study by the WHO on psychiatric disorders in general health care was carried out in 1990, 1 year after the opening of the Berlin Wall, in both parts of Berlin and in Mainz, West Germany. This allowed for a comparison of the prevalence rates of psychiatric disorders among general health care patients in the East and West, after the euphoria immediately following unification had subsided. RESULTS: The prevalence rates of current ICD-10 diagnoses and of subthreshold disorders in East Berlin were similar to the rates in West Berlin and Mainz. The recognition rate of psychiatric disorders by physicians did not differ in East Berlin as compared to West Berlin and Mainz. CONCLUSION: Contrary to the prediction expected from the literature on individual negative life events, major changes in life circumstances and stressful life events on a societal level within 1 year did not have a major impact on psychological function. PMID- 10365626 TI - Whom to ask for help in case of a mental disorder? Preferences of the lay public. AB - BACKGROUND: Although socio-cultural factors have been recognised as an important predictor in shaping help-seeking behaviour, few attempts have been made in this regard to specify the nature and impact of socio-cultural factors such as attitudes and belief systems prevalent in society. METHODS: We investigated the lay public's attitudes toward help-seeking regarding psychiatric disorders, and their determinants, in a cross-sectional national survey in Germany (n = 1564), using structured interviews with vignettes depicting a person either suffering from depression or from schizophrenia. Two distinct methodological approaches (rating vs ranking) were applied. RESULTS: Public opinion considers mental health professionals helpful in treating schizophrenia but not in the treatment of depression. For depression, public opinion clearly favours the lay support system and believes in involving the family physician if the former resource is exhausted. Determinants of help-seeking recommendations were problem definition, perception of the cause of distress and anticipated prognosis, as well as resentment against mental health professionals. CONCLUSION: Our results suggest that attitudes and belief systems prevalent in society have a major impact on help-seeking behaviour, both through transmission to the person suffering from mental distress via his/her social network and through the person's own attitudes formed in the process of socialization. Implications are pointed out for the daily work of mental health care providers, health care planning and public discussion of mental health issues. PMID- 10365627 TI - Factor structure of the CES-D (Center for Epidemiologic Studies Depression Scale) among Filipino-American adolescents. AB - The present study used factor analytic procedures to examine the factor structure of the CES-D among Filipino-American adolescents residing in rural and small town Hawaii. A total of 243 Filipino-American high school students completed the 20 item scale, and maximum likelihood analyses were employed to obtain a final solution. The results indicated that two factors provide a reasonably good fit: factor I combined depressed affect, somatic-retardation and interpersonal items, and factor II consisted of the remaining four positive affect items. The overlap of depressed affect and somatic symptoms support previous findings found among Asian American adults and other ethnic minority adolescents. The loading of the interpersonal items on the first factor is more unusual and suggests that interpersonal factors are not distinguished from depressed affect for the Filipino-American adolescent group. The usefulness of the CES-D as a tool to gain an understanding of the concept of depression across cultures is discussed. PMID- 10365628 TI - Social Support Questionnaire among psychiatric patients with various diagnoses and normal controls. AB - BACKGROUND: Several studies have pointed to the importance of social support in influencing the onset and course of a psychiatric disorder such as schizophrenia or depression. However, only a few have studied it across groups of patients with various psychiatric diagnoses employing a standardized assessment procedure. METHOD: We administered the Social Support Questionnaire (SSQ); a measure of social support recommended by two recent reviews on the subject, to 1,369 psychiatric outpatients visiting the 23 psychiatric hospitals and clinics all over Japan and to 178 healthy controls recruited from among employees at a general hospital. RESULTS: The original two-factor structure of the SSQ was confirmed and internal consistency reliability for the Number and Satisfaction subscales was satisfactory, with Cronbach's alphas above 0.85. When the SSQ scores were compared between psychiatric patients and healthy controls, it was found that the psychiatric patients in general reported significantly lower Number as well as Satisfaction scores than the healthy controls. When individual diagnostic categories were considered, almost all the diagnostic groups reported significantly lower Number scores, but only the patients with anxiety disorder, mood disorder, schizophrenia, and V codes reported significantly lower Satisfaction scores than the healthy controls. Compared with patients with other diagnoses, the schizophrenic patients stood out as reporting significantly lower Number and Satisfaction scores. CONCLUSION: The findings demonstrated the internal consistency reliability, factor validity, and construct validity of the SSQ among psychiatric as well as normal populations, and exemplified the feasibility of applying the SSQ as a standard measure of social support among psychiatric patients. PMID- 10365629 TI - The rationale, development and reliability of a new screening psychiatric instrument. AB - BACKGROUND: This paper describes the rationale, development, reliability and validity of a new screening psychiatric instrument. METHOD: The instrument comprises 26 items that tap the cardinal features of main psychiatric categories as defined by ICD-10 and DSM-IV. These items were adapted from various structured and semi-structured diagnostic interviews that yield ICD-10 and DSM-IV psychiatric diagnoses. After a training course, 12 trainees and the trainer rated blindly the 26 items on 45 subjects (22 with psychopathology and 23 without). Inter-rater reliability coefficient (Kappa) was estimated between trainees and the trainer on each item of the instrument. The total score on the new instrument was then correlated with the total score on the Arabic Self Reporting Questionnaire (SRQ-20) and the Arabic version of the General Health Questionnaire (GHQ) in a random sample from the general population (n = 365). Logistic regression was utilised to estimate the power of the total score on the new instrument in discriminating between cases and non-cases as classified by the SRQ 20. RESULTS: Excellent levels of agreement (Kappa > 0.80) were found for all items except for obsession (Kappa = 0.65) and for depressed mood (Kappa = 0.70). Moderate correlations were found between the total score on the new instrument and total score on SRQ-20 (r = 0.69) and the total score on the Arabic GHQ (r = 0.7). The new instrument correctly classified 89% of subjects into cases and non cases. CONCLUSIONS: The results of this study indicate that the new instrument is a highly reliable and valid screening instrument. The authors are now investigating its test-retest reliability and its procedural validity. PMID- 10365630 TI - Cluster of HIV-positive young women--New York, 1997-1998. AB - As of July 1997, six human immunodeficiency virus (HIV) infections in young women who reported sexual contact with the same HIV-infected man (putative index case patient) were detected at health-service clinics in a rural county in upstate New York. During the next several months, other sexual contacts of the man were discovered by public health officials through routine voluntary partner notification interviews, interviews with exposed women, and after a public announcement resulted in counseling and testing of approximately 1400 persons in the county. This report presents epidemiologic and laboratory findings of the young women investigated as part of this cluster and suggests a common source of HIV infection for these women. PMID- 10365631 TI - Progress toward global poliomyelitis eradication--1997-1998. AB - In 1988, the World Health Assembly resolved to eradicate poliomyelitis globally by 2000. Since then, substantial progress has been reported by all countries where polio is endemic in implementing the recommended polio eradication strategies (i.e., achieving and maintaining high routine coverage with oral poliovirus vaccine [OPV]; conducting National Immunization Days [NIDs] to rapidly decrease poliovirus circulation; establishing sensitive surveillance systems for polio cases and poliovirus; and carrying out mopping-up vaccination activities to eliminate the remaining reservoirs of poliovirus transmission). Although much progress has been made in many countries, substantial obstacles remain, particularly in 14 priority countries (i.e., global reservoir countries or countries with ongoing armed internal strife or civil war). This report updates progress during 1998 toward the global eradication target and describes accelerated activities to achieve the 2000 goal. PMID- 10365632 TI - Cigarette smoking during the last 3 months of pregnancy among women who gave birth to live infants--Maine, 1988-1997. AB - Cigarette smoking during pregnancy is associated with adverse birth outcomes (e.g., low birthweight and preterm delivery). The adverse effect of smoking on birthweight occurs primarily during the last trimester of pregnancy. To study smoking prevalence over time among women who gave birth to live infants in Maine, CDC and the Maine Department of Human Services (MDHS) analyzed self-reported data from the Pregnancy Risk Assessment Monitoring System (PRAMS) collected during 1988-1997. This report summarizes the results of this analysis, which indicate that despite the overall decline in smoking prevalence in Maine among women who gave birth to live infants, smoking prevalence remains high during the last 3 months of pregnancy among young women and low-income women, particularly those participating in the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC). PMID- 10365633 TI - Laboratory practices for prenatal Group B streptococcal screening and reporting- Connecticut, Georgia, and Minnesota, 1997-1998. AB - Group B Streptococcus (GBS) is a leading cause of neonatal sepsis in the United States. CDC, in collaboration with the American College of Obstetricians and Gynecologists and the American Academy of Pediatrics, recommends that laboratories adopt optimal screening practices to identify GBS and to promptly report test results so that GBS-colonized pregnant women can receive antibiotics during labor. To assess GBS screening practices in clinical laboratories, state health departments surveyed laboratories in Connecticut, Georgia, and Minnesota, participants in the Emerging Infections Program. The survey found that the practices of some participating laboratories were suboptimal, particularly in their lack of use of selective broth media for culture of GBS. PMID- 10365634 TI - Pharmacokinetic consequences of a citalopram treatment discontinuation. AB - In this pilot study, the pharmacokinetics of citalopram (CIT) were examined in five hospitalized depressed patients after an abrupt discontinuation of a treatment with 40 mg/d of this selective serotonin reuptake inhibitor (SSRI). During the 8-day study period, clinical ratings were regularly carried out. Between days 5 and 8, the patients were treated with clomipramine (75 mg/d). The enantiomers of CIT and its metabolites, demethyl-CIT (DCIT) and CIT-propionic acid derivative (CIT-PROP), were measured repeatedly from day 0 to day 8 by a stereoselective high-performance liquid chromatography (HPLC) procedure. The following drug plasma half-lives were measured (means +/- SD): R-CIT: 66+/-11 h; S-CIT: 42+/-13 h; R-DCIT: 228+/-148 h; S-DCIT: 93+/-35 h; R-CIT-PROP: 82+/-31 h; S-CIT-PROP: 186+/-93 h. PMID- 10365635 TI - The effect of diazepam in the recovery of rabbits from acute acetaminophen intoxication. AB - We have recently shown that diazepam can reduce mortality of acute iron overdose in rats. The mechanism for that effect is not yet defined. Our objective in the present study was to assess whether diazepam can similarly reduce mortality of experimental acute acetaminophen intoxication. Survival of rabbits was compared among four groups receiving 3 g/kg (body weight) of acetaminophen (LD40) orally each, followed by: 1) nothing (group I), 2) one oral dose of 140 mg/kg N acetylcystein (NAC) an hour later (group II), 3) intramuscular injection of 7 mg/kg diazepam (group III), 4) intramuscular injection of 7 mg/kg diazepam and one oral dose of 140 mg/kg NAC an hour later (group IV). 37.5% of rabbits in group I died after 16 hours, whereas none of the rabbits in group III died, (p = 0.04). No animal died during the 96-hour observation period in groups II and IV. Two and four hours post drug administration, acetaminophen plasma concentrations (APC) were significantly lower among rabbits in group III than in group I (p = 0.0007 and 0.01, respectively) and significantly lower among rabbits in group IV than in those in group II (p<0.0001 and p = 0.03, respectively). Acetaminophen plasma concentrations 2 hours after drug administration were also significantly lower among rabbits in group III than in those in group II (p = 0.0002). Seven and 24 hours after dosage, APC tended to be higher among rabbits in group III than in those in group I, but not significantly so. Administration of diazepam without NAC did not prevent liver and renal dysfunction. We conclude that early administration of diazepam in acute experimental acetaminophen overdose in rabbits reduced APC and mortality, probably by slowing intestinal motility, which resulted in delayed acetaminophen absorption from the gastrointestinal tract. PMID- 10365636 TI - Study of FK-binding protein: FK506-metabolite complexes by electrospray mass spectrometry: correlation to immunosuppressive activity. AB - Electrospray ionization mass spectrometry was used to study several non-covalent FK-binding protein (FKBP) immunosuppressant complexes in the gas phase. Relative FKBP binding affinities were determined from the signal ratio for the 7+ charge states of bound and unbound complexes as a function of capillary exit voltage. All complexes displayed a 1:1 binding stoichiometry. The relative gas-phase binding affinities were found to be well correlated with in vitro FKBP binding and in vitro immunosuppression (rapamycin > FK506 > or = 31-demethyl FK506 > 13 demethyl FK506 >> Cyclosporin A; CsA). The method demonstrates potential as a simple, rapid, and automatable technique for prediction of the immunosuppressive activity of FKBP:drug complexes. PMID- 10365637 TI - Population pharmacokinetics of aminoglycoside antibiotics in patients with cystic fibrosis. AB - The population pharmacokinetics of gentamicin and tobramycin were investigated in a group of 51 young adults with cystic fibrosis. Their ages ranged from 14-35 years, weights from 38-82 kg, and 27 of the patients were female. None of the patients had renal impairment, but 3 patients were treated in the intensive therapy unit (ITU) during one of their courses of therapy. Data comprised 219 courses of therapy and 544 concentrations (mean: 11 per patient). Concentration time data were analyzed using a nonlinear mixed-effects model package (NONMEM) and were best described by a one-compartment model. Factors identified as potentially influencing aminoglycoside pharmacokinetics were added in a stepwise fashion and the best model found that drug clearance and volume of distribution were related to body surface area and admission to ITU. The mean population estimates were a clearance of 2.89 L/hr/m2 and a volume of distribution of 9.21 L/m2 with a 60% increase in patients who were admitted to ITU. Interpatient variability in clearance and volume were 14% and 8%, respectively. The results suggested that a dose of 120 mg/m2 should achieve an average 1 hour postdose peak of 10 mg/L and trough of <1 mg/L and that higher doses might be required in ITU patients. PMID- 10365638 TI - Valproic acid toxicokinetics: serial hemodialysis and hemoperfusion. AB - The toxicity and pharmacokinetic properties of a drug determine whether hemodialysis and/or hemoperfusion are indicated in acute intoxications. Valproic acid is considered unremovable by hemodialysis because of the high protein binding of 90%-95%. A 27-year-old male with a history of seizures was admitted to the emergency room because of coma, hypernatriemia, and respiratory failure caused by an intoxication with a large dose of valproic acid. At admission, the plasma valproic acid level was 1414 mg/L (9.9 mmol/L) (therapeutic range: 50-100 mg/L (350-700 micromol/ L). The anion gap was 26 mmol/L (normal <12-14 mmol/L) and corresponded fairly well with this valproic acid level. Because of the potential toxicity of this high valproic acid level serial hemodialysis and hemoperfusion was performed. The first session was done with a charcoal column and the second session with a resin column. The patient recovered during the course of treatment. The valproic acid plasma clearances during treatment were: 80 mL/min (hemodialysis); 40 mL/min (hemoperfusion by charcoal) and 80 mL/min (hemoperfusion by resin, only in the first hour). The protein binding of valproic acid in plasma was only 32% at the start and was 54% at the end of the two sessions. In this specific case of a severe valproic acid intoxication, saturated protein binding resulted in an increased fraction of unbound valproic acid. This made hemodialysis an effective treatment, while hemoperfusion was relatively less effective because of saturation of the column. In conclusion, the toxicokinetics of valproate are quite different from the pharmacokinetics at therapeutic levels. The anion gap and protein binding are important parameters in toxicokinetics. PMID- 10365639 TI - Lower plasma levels of haloperidol in smoking than in nonsmoking schizophrenic patients. AB - The impact of smoking on plasma haloperidol (HAL) concentrations was investigated in 66 Japanese male schizophrenic inpatients treated orally with HAL. The subjects consisted of 22 nonsmokers and 44 smokers each smoking ten cigarettes per day. Plasma concentrations of HAL were determined by an enzyme immunoassay method. There were significant positive correlations between the plasma HAL concentration and the daily dose of HAL per kg body weight (Y = 58.1X-0.01 (r = 0.86)). Smokers had significantly lower HAL concentrations per daily dose of HAL/kg body weight than nonsmokers (smokers vs. nonsmokers = 54.3+/-16.6 vs. 70.6+/-23.2 ng/mL/mg/kg). In doses less than 0.2 mg/kg of HAL, smokers showed significantly lower HAL concentrations per daily dose of HAL/kg body weight than nonsmokers (smokers vs. nonsmokers = 55.1+/-14.4 vs. 79.5+/-27.1 ng/mL/mg/kg), whereas no significant difference in HAL concentrations per daily dose of HAL/kg body weight was observed between smokers and nonsmokers when treated with more than 0.2 mg/kg (smokers vs. nonsmokers = 52.9+/-20.7 vs. 60.0+/-11.1 ng/mL/mg/kg). Our results indicate that smoking may induce the enzyme(s) metabolizing HAL, which results in lower plasma HAL concentrations in smokers than in nonsmokers, particularly at low doses of HAL. PMID- 10365641 TI - Quinidine inhibition of cocaethylene degradation in human serum in vitro: a preliminary study. AB - Quinidine (QUINID) substantially inhibited the serum degradation of cocaethylene (CE), an important cocaine analog, when incubated for two hours at 37 degrees C with pooled human serum containing 1.58 micromol/L of CE. In the absence of QUINID, the CE concentration was only 49.4% of its original value after incubation. However, QUINID at subtherapeutic (3.08 micromol/L) and therapeutic (15.4 micromol/L) concentrations preserved 55.1% and 75.9% of the original CE concentration, respectively. At a QUINID concentration of 308 micromol/L, 89.2% of the CE concentration was preserved. There was a marked decrease in the serum cholinesterase (CHE) activity as QUINID concentrations were increased to 154 micromol/L (19.8% decrease in CHE) and to 308 micromol/L (36.8% decrease in CHE). These data suggest that QUINID inhibits the hydrolysis of CE in human serum by suppressing CHE activity. These findings are significant because QUINID may be used in patients who have abused cocaine with ethanol. They also have important implications for individuals receiving QUINID together with other drugs whose hydrolysis may be catalyzed by CHE. PMID- 10365640 TI - No pharmacokinetic but pharmacodynamic interactions between cisapride and bromperidol or haloperidol. AB - Pharmacokinetic and pharmacodynamic interactions between the gastrokinetic drug cisapride and the antipsychotic drugs bromperidol and haloperidol were studied in 29 schizophrenic inpatients. Fourteen patients were taking bromperidol (12-24 mg/d), and 15 were taking haloperidol (12-36 mg/d). Cisapride 10 mg/d was coadministered for 1 week, and blood sampling was performed before cisapride treatment, 1 week after starting cisapride treatment, and I week after stopping cisapride treatment. On the same days as the blood sampling, psychotic symptoms and side effects were evaluated using the Brief Psychiatric Rating Scale (BPRS) and the Udvalg for Kliniske Undersogelser (UKU) side effect rating scale (UKU), respectively. Plasma concentrations of bromperidol, haloperidol, and their reduced metabolites were measured by high-performance liquid chromatography. The mean BPRS scores after adding cisapride were significantly higher than those before cisapride (p<0.01) and after stopping cisapride (p<0.001) in the haloperidol group in this uncontrolled study. A similar tendency was observed in the bromperidol group, although it did not reach statistical significance (p = 0.08). Cisapride coadministration caused no significant changes in the mean plasma concentrations of bromperidol, haloperidol, and their reduced metabolites or in the mean UKU score. The present study suggests that there is no significant pharmacokinetic interaction between cisapride and bromperidol or haloperidol, but cisapride appears to deteriorate psychotic symptoms by a pharmacodynamic interaction in schizophrenic patients treated with haloperidol. PMID- 10365642 TI - Grapefruit juice decreases the systemic availability of itraconazole capsules in healthy volunteers. AB - The systemic availability of itraconazole capsules may be reduced secondary to elevated gastric pH and possibly by presystemic intestinal metabolism via CYP3A4. Grapefruit juice is acidic and an inhibitor of intestinal CYP3A4. To determine the effect of grapefruit juice on the systemic availability of itraconazole capsules, serum itraconazole and hydroxy-itraconazole concentrations were determined in eleven healthy volunteers studied in a randomized, two-way crossover design. Concurrent grapefruit juice resulted in a 43% decrease in the mean itraconazole AUC0-48 (2507 ng x hr/mL versus 1434 ng x hr/mL, p = 0.046) and a 47% decrease in the mean hydroxy-itraconazole AUC0-72 (7264 ng x hr/mL versus 3880 ng x hr/mL, p = 0.025). Grapefruit juice also significantly increased the mean itraconazole Tmax (5.5 versus 4 hours). We conclude that concomitant grapefruit juice does not enhance the systemic availability of itraconazole capsules, but rather appears to impair itraconazole absorption. Therefore, concomitant grapefruit juice will not likely be useful in improving the oral availability of itraconazole capsules. PMID- 10365643 TI - Pharmacokinetics and pharmacodynamics of the acetylcholinesterase inhibitor metrifonate in patients with renal impairment. AB - The aim of this study was to assess the influence of renal function on the pharmacokinetics, pharmacodynamics, safety, and tolerability of the acetylcholinesterase inhibitor metrifonate. Four groups of six age- and gender matched subjects with varying degrees of renal function (creatinine clearances more than 90, 60-90, 30-60, and less than 30 mL/min/ 1.73 m2, respectively) were administered a single 50-mg oral dose of metrifonate. Blood and urine samples were collected for 24 hours and concentrations of metrifonate and its metabolites dichlorvos, dichloroacetic acid, and M3 were determined. Inhibition of acetylcholinesterase activity in erythrocytes and butyrylcholinesterase in plasma were also measured. Metrifonate was well tolerated in all treatment groups. The urinary excretion of metrifonate and dichlorvos decreased with decreasing renal function but accounted for less than 2% of the elimination. There were no statistically significant differences in primary pharmacokinetic parameters- Cmax, t(max), area under the concentration-time curve (AUC), and t1/2--of metrifonate and dichlorvos among the different groups. The excretion of dichloroacetic acid and M3 was not influenced by renal impairment. Acetylcholinesterase was not inhibited, whereas butyrylcholinesterase was inhibited markedly but independently of renal function. No metrifonate dose adjustments are needed when treating subjects with renal impairment. PMID- 10365644 TI - Determination of buspirone and 1-(2-pyrimidinyl)-piperazine (1-PP) in human plasma by capillary gas chromatography. AB - Two separate gas chromatographic methods for the determination of buspirone and its active metabolite, 1-(2-pyrimidinyl)-piperazine (1-PP) in human plasma are described. Both procedures involve solid-phase extraction (the packing material of the cartridges used was C8 for buspirone and a mixed-mode sorbent for 1-PP), injection of the sample into a gas chromatograph equipped with a fused-silica capillary column and a nitrogen-phosphorus detector, and analysis with temperature programming (from 220 degrees C to 285 degrees C for buspirone and from 138 degrees C to 285 degrees C for 1-PP). The coating material of the analytical column was 5% diphenyl dimethyl silicone for buspirone and 50% diphenyl dimethyl silicone for 1-PP. Zolpidem was used as an internal standard in the buspirone assay and 1-phenylpiperazine in the 1-PP assay. Recovery of buspirone and 1-PP averaged 98% and 89%, respectively, and the limit of quantification was 0.2 ng/mL for both compounds. The between-run coefficients of variation ranged from 3.2% to 9.4% and from 2.9% to 8.6% for samples spiked with three different concentrations of buspirone and 1-PP, respectively. The suitability of these assays for pharmacokinetic studies was shown by analyzing timed plasma samples from volunteers after ingestion of a single therapeutic dose of buspirone (10 mg). PMID- 10365645 TI - A simple HPLC method for simultaneous determination of mycophenolic acid and mycophenolic acid glucuronide in plasma. AB - A reversed-phase high-performance liquid chromatographic method for the simultaneous determination of mycophenolic acid and its metabolite, mycophenolic acid glucuronide, is presented herein. Sample purification is limited to protein precipitation with acetonitrile. The analytes were separated on a C18 column with a mobile phase containing 30% acetonitrile and a 40 mm phosphoric acid buffer at pH 2.1 and measured with UV-detection at 215 nm. PMID- 10365646 TI - Measurement of mycophenolate mofetil plasma levels after heart transplantation and a potential side effect of high levels. AB - Mycophenolate mofetil (MMF) is gaining momentum in its use as an immunosuppressant and in the field of heart transplantation because of its efficacy and ease of use without a reported need to monitor plasma levels. We describe a case in which standard dosage of MMF (initially 1.5 g twice daily) produced elevated trough levels of mycophenolic acid (MPA). Although organ rejection was eradicated by the use of MMF, the patient developed severe anemia, which required repeated blood transfusions while the patient was on therapy. This case illustrates the potential value of monitoring MPA concentrations. PMID- 10365647 TI - Clozapine monitoring: plasma or serum levels? AB - In this study, differences between plasma and serum levels of clozapine and norclozapine were quantified and correlations of these differences with myeloperoxidase (MPO) activities were examined. Fifty-seven patients (34 men and 23 women, mean age 36.6+/-11.4 years) taking clozapine in a daily doses varying from 75 to 800 mg entered the study. All patients were analyzed for clozapine level, 29 were analyzed for norclozapine level and 28 for MPO activity. Significant differences (p<0.001) between plasma and serum levels were found for clozapine (mean decrease 9.99+/-15.8%) and norclozapine (mean decrease 16.0+/ 21.2%) and for MPO activities (mean increase 114.1+/-123.9%). No correlation was found between differences of MPO activities and differences of drug levels in plasma and serum. Differences in clozapine and norclozapine levels could reflect chemical instability of these drugs. These compounds should be quantified only in plasma because serum levels underestimate blood levels in patients. PMID- 10365648 TI - Determination of plasma metformin by a new cation-exchange HPLC technique. AB - Metformin is an oral antihyperglycemic agent used in the therapy of noninsulin dependent diabetic patients. This biguanide can induce dangerous complications such as lactic acidosis when its plasma concentration is too high. For this reason, the determination of plasma metformin should always be done during treatment. We developed a new HPLC method, for the routine determination of plasma metformin, with good reliability, rapid execution, and low costs. Sample preparation involved precipitation of the plasma proteins containing the internal standard buformin with a mixture of methanol, zinc sulfate, and ethylene glycol; the diluted supernatant was injected into a cation-exchange column. The mobile phase was potassium dihydrogenphosphate buffer-containing acetonitrile. The eluent was monitored at 236 nm. The calibration curve is linear within the range of 20-4000 ng/mL; the within-day coefficients of variation were less than 2.2% for metformin and 1.5% for buformin; the day-to-day coefficients of variation were less than 2.5% for metformin and 1.9% for buformin. The mean recoveries obtained from supplemented samples were included between 99.4 and 104.2% for metformin. Many characteristics make this method useful and easily accessible to all clinical laboratories equipped with HPLC instrumentation. PMID- 10365649 TI - Simultaneous determination of grepafloxacin, ciprofloxacin, and theophylline in human plasma and urine by HPLC. AB - A specific and sensitive reversed-phase high-performance liquid chromatographic (HPLC) method has been developed and validated for the simultaneous determination of grepafloxacin, ciprofloxacin, and theophylline in human plasma and urine. This assay allows these drugs to elute and be resolved in a single chromatogram at 280 nm, using a linear gradient. The procedure involves liquid-liquid extraction. Separation was achieved on a C18 reversed-phase column. The quantification limits were 0.05 mg/L in plasma and 0.5 mg/L in urine for grepafloxacin and ciprofloxacin and 0.5 mg/L in plasma and urine for theophylline. Standard curves were linear (correlation coefficients >0.999) over the ranges 0.05 to 5 mg/L for grepafloxacin and ciprofloxacin in plasma, from 0.5 to 20 mg/L for theophylline in plasma, and from 0.5 to 500 mg/L for the three drugs in urine. The coefficients of variation for the three drugs were less than 10% for within- and between-day analyses. The recoveries averaged 94.5% for theophylline, 93% for ciprofloxacin, 93.7% for grepafloxacin, and 95.1% for the internal standard (IS). The assay can be used for pharmacokinetic studies of these drugs, to investigate the interaction of grepafloxacin and ciprofloxacin with theophylline, or for routine simultaneous monitoring of theophylline, grepafloxacin, and ciprofloxacin. PMID- 10365650 TI - Small effects of valproic acid on the plasma concentrations of clozapine and its major metabolites in patients with schizophrenic or affective disorders. AB - Two separate studies were carried out to assess the effect of valproic acid on the steady-state plasma concentrations of clozapine and its major metabolites norclozapine and clozapine N-oxide in psychotic patients. In the first study, concentrations of clozapine and metabolites were compared between patients treated with clozapine in combination with sodium valproate (n = 15) and control patients treated with clozapine alone (n = 22) and matched for sex, age, body weight, and antipsychotic dosage. Patients comedicated with valproate tended to have higher clozapine levels and lower norclozapine levels, but the differences did not reach statistical significance. In a subsequent study, plasma concentrations of clozapine and its metabolites were determined in 6 patients with schizophrenia stabilized on clozapine therapy (200-400 mg/d) before and after treatment with sodium valproate (900-1200 mg/d) for 4 weeks. Mean plasma concentrations of clozapine and its metabolites did not change significantly throughout the study, but there was a trend for clozapine levels to be higher and for norclozapine levels to be lower after valproate. Overall, these findings suggest that valproic acid may have an inhibiting effect on the CYP1A2- or CYP3A4 mediated conversion of clozapine to norclozapine. However, the interaction is unlikely to be clinically significant. PMID- 10365651 TI - High-performance liquid chromatography method for analyzing the antiretroviral agent efavirenz in human plasma. AB - Efavirenz (EFV, DMP-266) is a new antiretroviral agent belonging to the class of nonnucleoside reverse transcriptase inhibitors. It has recently been approved by the Food and Drug Administration in management of human immunodeficiency virus (HIV). Preliminary pharmacokinetic studies on EFV in healthy volunteers show that the drug may influence the metabolism of protease inhibitors. For the determination of EFV in human plasma, a validated and specific reverse-phase high performance liquid chromatography (HPLC) method, with UV detection, was developed. We used 100 microL plasma sample for a liquid-liquid extraction with diethyl ether after basification. The mobile phase was a mixture of acetonitrile and water, pumped at a flow rate of 1.2 mL/min. Ultraviolet detection was carried out at a wavelength of 247 nm. Retention times for EFV and internal standard (IS) were 5.3 and 4.5 minutes, respectively, and there was no chromatographic interference from other commonly administered drugs. The limit of detection was 100 ng/mL. The described assay is a rapid and accurate method for measurement of EFV in plasma: the easy preparation and small sample size makes this assay highly suitable for pharmacokinetic studies and routine clinical analysis in patients with HIV. In addition, the reproducibility of the method is only moderately increased by including IS, so analyzing without IS may be an alternative. PMID- 10365652 TI - Prediction of valproate serum concentrations in adult psychiatric patients using Bayesian model estimations with NPEM2 population pharmacokinetic parameters. AB - Valproate serum concentrations between 45 and 125 microg/mL are associated with the drug's efficacy in acute mania. Adaptive control dosing of valproate has not been fully studied in psychiatry. The objective of this study was to derive population pharmacokinetic (PK) parameters for valproate in healthy volunteers and to test the ability of these PK parameters to estimate concentrations in adult psychiatric patients using a Bayesian program. Population PK parameters for oral valproate were estimated from 18 PK studies in six healthy volunteers (1) using NPEM2. A Bayesian PK program using these population parameters was used to predict valproate concentration-time points in a second cohort of 21 adult psychiatry patients using 0, 1, or 2 prior concentrations. Estimated population parameters (mean +/- SD) were: Ka, 1.15+/-1.75/h; V, 0.14+/-0.042 L/Kg; and CL, 0.902+/-0.133 L/h. Bayesian valproate estimations using these parameters were negatively biased (underestimations) using zero prior concentration and unbiased using 1 or 2 prior concentrations. Mean error values (95% CI) in microg/mL for predictions using 0, 1, or 2 prior concentration-time points were -12.0 (-22.5, 1.5), -9.5 (-19.1, 0.1), and -2.5 (-11.1, 6.1), respectively, and mean absolute error values in microg/mL (95% CI) were 19.8 (12.6, 27.1), 16.3 (9.4, 23.3), and 10.1 (4.9, 15.2), respectively. Population parameters derived from healthy adult volunteers provided biased predictions of valproate concentrations in adult psychiatric patients. However, estimates using 1 or 2 valproate concentration time points predicted future concentrations that were precise and unbiased, given the wide therapeutic target range. PMID- 10365653 TI - Determination of mitotane (o,p-DDD) and its metabolites o,p-DDA and o,p-DDE in plasma by high-performance liquid chromatography. AB - The adrenolytic agent mitotane (o,p'-DDD or 1,1-(o,p'-dichlorodiphenyl)-2,2 dichoroethane) is believed to be metabolically activated, resulting in the generation of reactive intermediates or the formation of o,p'-DDA. A new high performance liquid chromatographic (HPLC) method for measurement of mitotane and two of its metabolites, o,p'-DDA (1,1-(o,p'-dichlorodiphenyl) acetic acid) and o,p'-DDE (1,1-(o,p'-dichlorodiphenyl)-2,2 dichloroethene), in plasma has been developed. Sample preparation involves precipitation of plasma proteins with acetone before chromatographic separation under isocratic conditions on a silica based diphenyl column. Mitotane and metabolites are quantified by ultraviolet detection at 218 nm. Recoveries for the three compounds range between 99% and 110%, with between-day and within-day coefficients of variation <6% within the therapeutic range. Limits of detection are 0.5 microM and the assay is linear up to at least 800 microM for o,p'-DDA and 100 microM for mitotane and o,p'-DDE. The method has been evaluated using samples from three patients on mitotane therapy, revealing o,p'-DDA levels in plasma 3 to 10 times higher than the levels of the parent compound. PMID- 10365654 TI - Determination of indinavir, a HIV-1 protease inhibitor, in human plasma using ion pair reversed-phase high-performance liquid chromatography. AB - Indinavir is widely prescribed as a component of potent antiretroviral therapy for the treatment of HIV-1 infection. Because virologic failure of therapy can result from subtherapeutic drug levels, monitoring of indinavir levels may be important in clinical management. We have developed a simple, accurate, and precise high-performance liquid chromatographic (HPLC) assay for measurement of indinavir concentration in human plasma. In our method, indinavir was extracted from plasma with dichloromethane at pH 10.4, which resulted in quantitative recovery of indinavir and the internal standard (IS), methyl-indinavir (86% and 80%-97%, respectively). Chromatographic separation was accomplished using a Luna C18 (2) (Phenomenex) analytic column with a mobile phase composed of acetonitrile:phosphate buffer (25 mM) and 0.2% triethylamine pH 7.0 (34.5:65.5, v/v). Ion-paired reagent triethylamine was necessary to ensure an appropriate retention time for indinavir and differentiate it from other protease inhibitors that were coextracted. Quantification was performed at 210 nm. The standard curves were linear (r2>0.999) over the concentration range 25-5,000 ng/mL, when 1 mL aliquots of plasma were extracted. Inter- and intraday coefficients of variation were acceptable. The assay was used to determine trough and peak levels of in plasma from 12 subjects who received indinavir 1200 mg every 12 hours, 1000 mg every 12 hours, or 800 mg every 8 hours. The concentrations of indinavir found in this study (trough 26-768 ng/mL; peak at 1 hr 3,309-17,568 ng/mL) has a wider range than defined previously (trough 50-300 ng/mL; peak 6,000-12,000 ng/mL). This study illustrates three potential uses of indinavir monitoring: to assess individual dosing regimen, to assess patient compliance, and to monitor unusual indinavir levels caused by changed drug clearance. PMID- 10365655 TI - An improved HPLC method for therapeutic drug monitoring of daunorubicin, idarubicin, doxorubicin, epirubicin, and their 13-dihydro metabolites in human plasma. AB - A single high-performance liquid chromatography (HPLC) method, suitable for the analysis of daunorubicin, idarubicin, doxorubicin, epirubicin, and their 13 dihydro metabolites is validated in the present study. Preparation of plasma samples was performed by a first extraction of analytes with a chloroform/1 heptanol mixture (9:1) and reextraction with ortophosphoric acid 0.1 M. The chromatographic analysis was carried out by reversed-phase isocratic elution of anthracyclines with a Supelcosil LC-CN 5 mm column (25 cm x 4.6 mm internal diameter; Supelco) and detection was accomplished by spectrofluorimetry at excitation and emission wavelengths of 480 and 560 nm, respectively. All anthracyclines eluted within 15 minutes of injection and the method appeared to be specific, because the chromatographic assay did not show interferences at the retention time of analytes. The linearity, evaluated over a concentration range of 0.4-10,000 ng/mL, gave regression coefficients better than 0.999, with recoveries of doxorubicin-doxorubicinol and epirubicin-epirubicinol of 67%-109% and 61%-109% respectively, and 93%-109% for the other compounds. The limits of detection and quantification were 0.4 ng/mL in a 50-mL sample (40 pg/injection) for all anthracyclines tested. The method proved to be precise and accurate, as the within-day and between-day coefficients of variation were less than 10% and the accuracy of the assay was in the range of 91%-107%. Overall results indicate that it is feasible to analyze all the anthracyclines used in clinical practice and their major metabolites with a single optimized method, thereby simplifying their monitoring in chemotherapeutic regimens of cancer patients. PMID- 10365656 TI - Rapid screening test for gamma-hydroxybutyric acid (GHB, Fantasy) in urine. PMID- 10365657 TI - Gene therapy with autologous, interleukin 2-secreting tumor cells in patients with malignant melanoma. AB - We vaccinated metastatic melanoma patients with irradiated, autologous melanoma cells genetically engineered to secrete interleukin 2 (IL-2) to investigate whether an anti-tumor immune response would be induced. Melanoma cell cultures were established from surgical specimens and were engineered to secrete IL-2 by infection with recombinant retrovirus. Twelve patients were vaccinated subcutaneously one, two, or three times with approximately 10(7) irradiated, autologous, IL-2-secreting tumor cells. Treatment was well tolerated, with local reactions at 11 of 24 injection sites and minor systemic symptoms of fever and headache after 6 injections. One patient developed anti-tumor DTH after the first vaccination and showed an increased response after the second vaccination. Anti autologous tumor CTLs could be detected prevaccination in the peripheral blood of seven patients and their activity increased after vaccination in four patients. No UICC-defined clinical responses were seen, but three patients had stable disease for 7-15 months, one of whom has not yet progressed (15+ months). Thus, patient vaccination with autologous, genetically engineered tumor cells is feasible and safe. Anti-tumor DTH and CTLs can be induced in some patients with such a vaccine. PMID- 10365658 TI - Cardiac resistance to adriamycin in transgenic mice expressing a rat alpha cardiac myosin heavy chain/human multiple drug resistance 1 fusion gene. AB - Cardiac toxicity is a major factor that limits the use of anthracyclines in cancer chemotherapy. Heart failure frequently develops in patients treated with doxorubicin (Adriamycin), when they receive a cumulative dose greater than 500 mg/m2. To make a mouse model for gene therapy designed to prevent this toxic effect, we have produced transgenic mice overexpressing the human cDNA for the multiple drug resistance (h-mdr1) gene driven by 2.12 kb of the 5' flanking region of the rat alpha-cardiac myosin (aCM) heavy chain gene. Two lines of transgenic mice expressed the transgene at a high level in heart muscle. Transgenic and control animals were treated with Adriamycin intravenously at either a single dose of 10 mg/kg or a cumulative dose of 30 mg/kg in three injections. Subsequent light and electron microscopic examination of heart tissue demonstrated degenerative changes in control mice that were absent in transgenic animals at both doses. These results show that expression of the alphaCM/h-mdr1 transgene in heart confers protection from the toxic effect of Adriamycin and suggest that such constructs, if employed effectively in cardiac gene therapy protocols, could allow a more aggressive use of anthracyclines in the treatment of cancer. PMID- 10365659 TI - Persistent delivery of factor IX in mice: gene therapy for hemophilia using implantable microcapsules. AB - Severe hemophilia B is a life-threatening, life long condition caused by absence of or defective coagulation factor IX. Gene therapy could provide an alternative treatment to repeated injection of plasma-derived concentrate or recombinant factor IX. We have previously described the use of implantable microcapsules containing recombinant myoblasts to deliver human factor IX in mice. This study reports the generation of improved myoblast-specific expression vectors. Mouse myoblast clones transfected with the various vectors secreted factor IX in vitro, at rates between 70 and 1000 ng/10(6) cells/day. The recombinant myoblast clones were then encapsulated and implanted into mice. Immunocompetent mice implanted with encapsulated myoblasts had up to 65 ng of factor IX per milliliter in their plasma for up to 14 days, after which antibodies to human factor IX became detectable, and this coincided with decreased factor IX in mouse plasma. In immunodeficient mice, however, factor IX delivery was maintained at a constant level for at least 6 weeks (end of experiment). Interestingly, the highest secreting myoblast clone in vitro did not deliver the highest level of hFIX in vivo. This discrepancy observed between performance in vitro and in vivo may have important implications for the development of gene therapy protocols based on recombinant cells. PMID- 10365660 TI - Phase I trial of interferon gamma retroviral vector administered intratumorally with multiple courses in patients with metastatic melanoma. AB - The purpose of this study was to determine the safety and antitumor activity of IFN-gamma retroviral vector in patients with advanced melanoma. Seventeen patients (9 single courses, 8 multiple courses) received a total of 363 intratumor injections of IFN-gamma retroviral vector (1 x 10(7) PFU/ml administered at 0.3, 0.5, and 1.0 ml per cohort). No grade III/IV adverse events were attributed to study medication. Replication-competent retrovirus was not detected in any of the 17 patients by polymerase chain reaction studies. Eight patients showed elevated anti-tumor antibody responses in comparison with baseline by ELISA. One of nine patients treated with a single course had an optimal response of stable disease, compared with eight of eight multiple injected patients. Median survival of single-injected patients was 150 days, and patients who received multiple injections have still not achieved median survival duration, with four of eight still living (p = 0.0462, Wilcoxon; p = 0.0273, log rank). We conclude that intratumor injection of IFN-gamma is safe and well tolerated. Evidence of antitumor activity is suggested in patients with advanced malignancy that received multiple injections. PMID- 10365661 TI - Adenovirus-mediated utrophin gene transfer mitigates the dystrophic phenotype of mdx mouse muscles. AB - Utrophin is a close homolog of dystrophin, the protein whose mutations cause Duchenne muscular dystrophy (DMD). Utrophin is present at low levels in normal and dystrophic muscle, whereas dystrophin is largely absent in DMD. In such cases, the replacement of dystrophin using a utrophin gene transfer strategy could be more advantageous because utrophin would not be a neoantigen. To establish if adenovirus (AV)-mediated utrophin gene transfer is a possible option for the treatment of DMD, an AV vector expressing a shortened version of utrophin (AdCMV-Utr) was constructed. The effect of utrophin overexpression was investigated following intramuscular injection of this AV into mdx mice, the mouse model of DMD. When the tibialis anterior (TA) muscles of 3- to 5-day-old animals were injected with 5 microl of AdCMV-Utr (7.0 x 10(11) virus/ml), an average of 32% of fibers were transduced and the transduction level remained stable for at least 60 days. The presence of utrophin restored the normal histochemical pattern of the dystrophin-associated protein complex at the cell surface and resulted in a reduction in the number of centrally nucleated fibers. The transduced fibers were largely impermeable to the tracer dye Evans blue, suggesting that utrophin protects the surface membrane from breakage. In vitro measurements of the force decline in response to high-stress eccentric contractions demonstrated that the muscles overexpressing utrophin were more resistant to mechanical stress-induced injury. Taken together, these data indicate that AV-mediated utrophin gene transfer can correct various aspects of the dystrophic phenotype. However, a progressive reduction in the number of transduced fibers was observed when the TA muscles of 30- to 45-day-old mice were injected with 25 microl of AdCMV-Utr. This reduction coincides with a humoral response to the AV and transgene, which consists of a hybrid mouse-human cDNA. PMID- 10365662 TI - Retrovirus vector-mediated correction and cross-correction of lysosomal alpha mannosidase deficiency in human and feline fibroblasts. AB - Lysosomal alpha-mannosidase (EC 3.2.1.24) is an exoglycosidase in the glycoprotein degradation pathway. A deficiency of this enzyme causes the lysosomal storage disease alpha-mannosidosis. Retrovirus vector transfer of a new human alpha-mannosidase cDNA resulted in high-level expression of alpha mannosidase enzymatic activity in deficient human and feline fibroblasts. The expressed alpha-mannosidase had the same biochemical properties (thermal stability, pH profile, inhibitor/activator sensitivity) as the native enzyme expressed in normal cells. The transferred enzyme colocalized with a control lysosomal hydrolase in cell fractionation experiments. The vector-encoded enzyme also was released at high levels from the corrected cells, and was taken up by untreated mutant cells via the mannose 6-phosphate receptor-mediated endocytic pathway (cross-correction). It is envisioned that genetic correction of a subset of cells (e.g., hematopoietic stem cells) in patients will provide a source of corrective enzyme for other affected tissues in this multisystem disease. Development of a vector expressing high levels of alpha-mannosidase that cross corrects mutant cells will enable somatic gene transfer experiments in the cat model of human alpha-mannosidosis. PMID- 10365663 TI - Retrovirus-mediated correction of the metabolic defect in cultured Farber disease cells. AB - Farber disease is a rare severe lysosomal storage disorder due to a deficient activity of the enzyme acid ceramidase (AC). Patients have granulomas along with lipid-laden macrophages that accumulate in a number of tissues, leading to multiple diverse clinical symptoms. There is no therapy for the disorder and most patients succumb to the disease in early childhood. The severity of the disease progression seems to correlate with the amount of the accumulated ceramide substrate. Since the cDNA for human AC has been elucidated we sought to establish if genetic transfer of this sequence would lead to enzymatic and, especially, functional correction of the catabolic defect in Farber patient cells. To do this, a novel amphotropic recombinant retrovirus was constructed that engineers transfer of the human AC cDNA. On infection of patient fibroblasts, AC enzyme activity in cell extracts was completely restored. Further, substrate-loading assays of intact living cells showed a fully normalized catabolism of lysosomal ceramide. Lastly, as reported for some other corrected enzymatic defects of lysosomes, metabolic cooperativity was seen, in that functionally corrected patient fibroblasts secreted AC that was taken up through the mannose 6-phosphate receptor and used by uncorrected fibroblasts as well as recipient Farber lymphoblastoid cells. This overall transduction and uptake scenario for Farber disease allows future treatment of this severe disorder to be envisioned using gene transfer approaches. PMID- 10365664 TI - Safety of direct myocardial administration of an adenovirus vector encoding vascular endothelial growth factor 121. AB - A gene therapy strategy involving direct myocardial administration of an adenovirus (Ad) vector encoding the vascular endothelial growth factor 121 cDNA (Ad(GV)VEGF121.10) has been shown to be capable of "biological revascularization" of ischemic myocardium in an established porcine model [Mack, C.A. (1998). J. Thorac. Cardiovasc. Surg. 115, 168-177]. The present study evaluates the local and systemic safety of this therapy in this porcine ischemia model and in normal mice. Myocardial ischemia was induced in Yorkshire swine with an ameroid constrictor 21 days prior to vector administration. Ad(GV)VEGF121.10 (10(9) or 10(10) PFU), Ad5 wild type (10(9) PFU), AdNull (control vector with no transgene; 10(9) PFU), saline, or no injection (naive) was administered in 10 sites in the ischemic, circumflex distribution of the myocardium. Toxicity was assessed by survival, serial echocardiography, blood analyses, and myocardial and liver histology at 3 and 28 days after vector administration. All pigs survived to sacrifice, except for one animal in the Ad(GV)VEGF121.10 (10(10) PFU) group, which died as a result of oversedation. Echocardiograms of Ad(GV)VEGF121.10 treated pigs demonstrated no differences in pericardial effusion, mitral valve regurgitation, or regional wall motion compared with control pigs. Intramyocardial administration of Ad(GV)VEGF121.10 included only minimal myocardial inflammation and necrosis, and no hepatic inflammation or necrosis. Only a mild elevation of the white blood cell count was encountered on day 3, which was transient and self-limited in the Ad(GV)VEGF121.10 group as compared with the saline-treated animals. As a measure of inadvertent intravascular administration of vector, normal C57/BL6 mice received intravenous Ad(GV)VEGF121.10 (10(4), 10(6), 5 x 10(7), or 10(9) PFU), AdNull (5 x 10(7) or 10(9) PFU), or saline. Toxicity was assessed by survival, blood analyses, and organ histology at 3 and 7 days after vector administration. A separate group of C57/BL6 mice received intravenous AdmVEGF164 (Ad vector encoding the murine VEGF164 cDNA), Ad(GV)VEGF121.10, AdNull (10(8) PFU each group), or saline to assess duration of expression and safety of a homologous transgene. All mice survived to sacrifice except for 40% of the mice in the highest (10(9) PFU; a dose more than 10(3)-fold higher by body weight than the efficacious dose in pigs) Ad(GV)VEGF121.10 dose group, which died on days 5-6 after vector administration. The only differences seen in the blood analyses between treated and control mice were in the very high Ad(GV)VEGF121.10 dose group (10(9) PFU), which demonstrated an anemia as well as an increase in alkaline phosphatase when compared with all other treatment groups. Hepatic VEGF levels by ELISA in AdmVEGF164-treated mice did not persist beyond 14 days after vector administration, suggesting that persistent expression of a homologous VEGF gene transferred with an Ad vector is not a significant safety risk. Although this is not a chronic toxicity study, these data demonstrate the safety of direct myocardial administration of Ad(GV)VEGF121.10, and support the potential use of this strategy to treat human myocardial ischemia. PMID- 10365665 TI - PEGylation of adenovirus with retention of infectivity and protection from neutralizing antibody in vitro and in vivo. AB - Replication-defective recombinant adenovirus (Ad) vectors are under development for a wide variety of gene therapy indications. A potential limiting factor associated with virus gene therapy requiring repeated treatment is the development of a humoral immune response to the vector by the host. In animal models, there is a dose-dependent rise in neutralizing antibodies after primary vector administration, which can preclude effective repeat administration. The strategy we have developed to circumvent the neutralization of adenovirus vectors by antibodies is to mask their surface by covalent attachment of the polymer polyethylene glycol (PEG). Covalent attachment of PEG to the surface of the adenovirus was achieved primarily by using activated PEG tresylmonomethoxypolyethylene glycol (TMPEG), which reacts preferentially with the epsilon-amino terminal of lysine residues. We show that the components of the capsid that elicit a neutralizing immune response, i.e., hexon, fiber, and penton base, are also the main targets for PEGylation. Several protocols for PEGylation of an adenovirus vector were evaluated with respect to retention of virus infectivity and masking from antibody neutralization. We show that covalent attachment of polymer to the surface of the adenovirus can be achieved with retention of infectivity. We show further that PEG-modified adenovirus can be protected from antibody neutralization in the lungs of mice with high antibody titers to adenovirus, suggesting that PEGylation will improve the ability to administer Ad vectors on a repeated basis. PMID- 10365666 TI - Characterization of genetically altered, interleukin 2-independent natural killer cell lines suitable for adoptive cellular immunotherapy. AB - NK-92 is a highly cytotoxic natural killer (NK) tumor cell line that possesses properties that make it an excellent candidate for adoptive cellular immunotherapy. However, the cytotoxicity of NK cells is dependent on cytokines such as interleukin 2 (IL-2). Although NK-92 cells maintain cytotoxicity for a time after withdrawal of IL-2, clinical use will probably require prolonged treatment with fully activated cells to eliminate disease effectively. The ability to support cytotoxic cells with exogenously administered IL-2 is limited by associated toxicity. Therefore, we describe the transfection of the IL-2 dependent NK-92 cell line with human IL-2 (hIL-2) cDNA by particle-mediated gene transfer to create two IL-2-independent variants, NK-92MI and NK-92 CI, and describe their characterization and comparison with parental cells. Both variants were shown to contain, express, and synthesize the hIL-2 cDNA. IL-2 synthesis was higher in NK-92MI cells compared with NK-92CI cells, with no expression in parental cells. Functionally, the cytotoxicity of all three cell lines was similar and coincubation with IL-2-independent variants did not affect hematopoietic progenitor cells. NK-92MI and NK-92CI cells were more radiosensitive than NK-92 cells, with proliferation inhibited at lower radiation doses and increased morality and decreased cytotoxicity compared with parental cells. Data presented here show that we have created by particle-mediated gene transfer two IL-2-independent variants of NK-92 that are identical to parental cells in virtually all respects, including high cytotoxic activity. The nonviral transfection of these cells makes them suitable for clinical applications. These IL-2-independent cells should allow prolonged treatment with fully active natural killer cells without the need for exogenous IL-2 support. PMID- 10365667 TI - Anti-tumor immunity induced by in vivo adenovirus vector-mediated expression of CD40 ligand in tumor cells. AB - CD40 ligand (CD40L), the ligand for CD40 on antigen-presenting cells, is essential for the initiation of antigen-specific T cell responses, an important component of the immune response to tumors. This study is based on the hypothesis that in vivo genetic modification of tumor cells to express CD40L will trigger CD40 on local antigen-presenting cells to present tumor antigen to the cellular immune systems, thus eliciting anti-tumor immunity to suppress growth of the tumor. To examine this concept, subcutaneous tumors of three different murine tumor models in two strains of mice were infected with a recombinant adenovirus (Ad) vector expressing murine CD40L (AdmCD40L). In the B16 (H-2b, melanoma) and CT26 (H-2d, colon cancer) murine models, injection of AdmCD40L into established subcutaneous tumors resulted in sustained tumor regression and tumor-free status in >60% of animals. Intratumoral injection of AdmCD40L also significantly suppressed the growth of established, weakly immunogenic Lewis lung carcinoma (H 2b) tumors, but to a lesser extent. Ex vivo AdmCD40L-transduced tumor cells implanted in syngeneic hosts induced significant antitumor response against preexisting identical tumors at a distant site. Both in vivo and in vitro AdmCD40L modification of tumors to express CD40L elicited tumor-specific cytolytic T lymphocytes responses, and the transfer of spleen cells from treated mice efficiently protected naive mice against a subsequent tumor challenge. These results support the concept that transduction of tumors with a recombinant CD40L adenovirus vector may be a useful strategy for cancer immunotherapy. PMID- 10365668 TI - Effect of scaffold attachment region on transgene expression in retrovirus vector transduced primary T cells and macrophages. AB - The scaffold attachment region of the human interferon beta gene (IFN-SAR) inserted into a retroviral vector improved transgene expression in human primary CD4+ and CD8+ T cells, and in primary monocytemacrophages. In T cells, expression of the Maloney murine leukemia virus (Mo-MuLV)-based retroviral vectors was high in activated cells but low in resting cells. Addition of the IFN-SAR sequence enhanced vector expression 2- to 10-fold, and the effect was particularly pronounced in resting T cells. In CD33+CD14+CD4+ monocyte-macrophages derived from transduced hematopoietic stem/progenitor cells (HSPCs) in vitro, the IFN-SAR enhanced vector expression three- to sixfold. We have used the IFN-SAR-containing vectors to express the RevM10 gene, a trans-dominant mutant of the human immunodeficiency virus type 1 (HIV-1) rev gene. Compared with a standard retroviral vector, the IFN-SAR-containing vector was significantly (p < 0.01) more potent at inhibiting HIV-1 replication in infected CD4+ peripheral blood lymphocytes. In monocytes, however, addition of the IFN-SAR did not significantly improve antiviral efficacy. To understand better the reason for the strong effect of the SAR on antiviral efficacy in T cells we have studied the expression of HIV, Mo-MuLV, and Mo-MuLV + SAR vectors in resting and activated cells. While the expression of all three vectors was lower in resting compared with activated cells, the kinetics of the decrease in expression were fastest for the Mo-MuLV vector, followed by the HIV vector and then the Mo-MuLV + SAR vector. Thus, higher level expression of the Mo-MuLV + SAR vector relative to wild-type HIV at all stages of T cell activation is the most likely explanation for the strong antiviral efficacy. Overall, this study demonstrates the utility of the IFN-SAR sequence for achieving high-level retroviral vector expression in lymphoid and myeloid hematopoietic cells. PMID- 10365670 TI - Recombinant human thyrotropin. PMID- 10365669 TI - A marker study of therapeutically transduced CD4+ peripheral blood lymphocytes in HIV discordant identical twins. PMID- 10365671 TI - An overview of the management of papillary and follicular thyroid carcinoma. AB - Long-term survival rate for papillary and follicular carcinoma is more than 90%, but this varies considerably among subsets of patients. About 30% of patients, however, develop tumor recurrence, depending on the initial therapy. Two-thirds of the recurrences occur within the first decade after therapy, but the others may appear years later. We found that among patients with recurrent cancer, 30% could not be fully eradicated and another 15% died of disease. Tumor recurred outside the neck in 21% of our patients, most commonly in the lungs (63%), which resulted in death in about half the patients. Mortality rates are lower when recurrences are detected early by radioiodine scans rather than by clinical signs. We believe that the best treatment for most patients with differentiated thyroid carcinoma is near-total thyroidectomy followed by 131I ablation of the thyroid remnant, which in our experience reduces the recurrence rate, improves survival and facilitates follow-up. A long delay in initiating this therapy has an adverse and independent effect on prognosis, more than doubling the 30-year cancer mortality rate. If only partial lobectomy has been performed, it is best to consider completion thyroidectomy for lesions 1 cm or larger because of the high rate of residual carcinoma in the contralateral lobe. Completion thyroidectomy and 131I whole-body scanning allows for the diagnosis and treatment of unrecognized carcinoma and when performed early, results in significantly fewer lymph node and hematogenous recurrences and enhances survival. A large and growing number of studies demonstrates decreased recurrence of papillary carcinoma and decreased disease-specific mortality attributable to 131I therapy. On the basis of our observations and other studies, we believe that an aggressive approach to initial management and follow-up may render nearly 90% of the patients permanently free of disease. Periodic follow-up should be done with whole-body scanning and serum thyroglobulin (Tg) measurements, performed either during thyroid hormone withdrawal or by recombinant human thyrotropin (TSH) stimulated scanning and Tg measurement. A scheme for follow-up management is presented. PMID- 10365672 TI - Strategies for thyrotropin use to monitor patients with treated thyroid carcinoma. AB - Detection of residual and recurrent thyroid carcinoma requires long-term monitoring of patients with serum thyroglobulin measurement and radioiodine scanning during temporary thyrotropin (TSH) stimulation. Recombinant thyrotropin (rTSH) permits these studies to be performed without the morbidity associated with withdrawal of thyroid hormone therapy. A protocol for rTSH use is proposed, beginning with measurement of serum thyroglobulin during TSH suppression. Patients at significant risk of recurrence with a low initial thyroglobulin level then have rTSH stimulation testing. Patients with positive rTSH-stimulated thyroglobulin concentrations and/or radioiodine scans can then be directed for appropriate therapy. The previously studied 2-dose rTSH protocol with imaging at 48 hours after 131I dosing requires Monday-through-Friday testing in most settings, but new regimens may be established. rTSH-stimulated testing may be less accurate in patients with thyroglobulin autoantibodies and those with residual normal thyroid tissue, and is generally unnecessary when there is other evidence of residual disease. Physicians should consider patients' pretest probability of disease in deciding whether and how often to perform rTSH stimulated testing after primary treatment for thyroid carcinoma. PMID- 10365673 TI - Detection of residual and recurrent differentiated thyroid carcinoma by serum thyroglobulin measurement. AB - Thyroglobulin (Tg) measurement is primarily used to monitor patients with differentiated thyroid carcinoma (DTC) for tumor recurrence. Serum Tg levels principally integrate 3 variables: the mass of thyroid tissue present (benign or neoplastic); the degree of thyrotropin (TSH) receptor stimulation and tumor's intrinsic ability to synthesize and secrete Tg--a factor that needs to be assessed by a preoperative serum Tg determination. Serum Tg measurements should be interpreted relative to the TSH status of the patient. When TSH is low (on levothyroxine [LT4] therapy) basal serum Tg may be undetectable and recombinant human thyrotropin (rhTSH) administration may be needed to increase serum Tg into the measureable range. The Tg fold response to rhTSH (rhTSH-stimulated Tg/basal Tg) is an index of the tumor's sensitivity to TSH. Normal thyroid remnant and well-differentiated thyroid tumors display a greater (>10-fold) serum Tg response to TSH stimulation compared with less well-differentiated tumors (<3-fold). The factors influencing the response include the magnitude and chronicity of the serum TSH elevation, the mass of thyroid tissue and the TSH receptor status of the tumor. Technical problems still compromise the clinical utility of serum Tg measurement. Thyroglobulin autoantibodies are present in approximately 20% of all DTC patients and cause either underestimation or overestimation of serum Tg measurements made by immunometric assay (IMA) and radioimmunoassay (RIA) methods, respectively. Other technical problems include poor interassay precision, "hook" effects (IMA methods), intermethod standardization differences, and suboptimal sensitivity for detecting small amounts of tumor during TSH suppression. When TSH is suppressed, the basal serum Tg provides an integrated index of thyroid tissue mass and its capability to secrete Tg. Serial measurements of basal Tg concentrations can be used to monitor tumor progression or regression. The development of a low (<1 ng/mL) serum Tg (on LT4 therapy) by the second postoperative year signifies a low 5-year recurrence risk whereas a rising serum Tg in the face of TSH suppression is an abnormal response consistent with recurrence. The optimal degree of TSH suppression for a patient should be based on clinical judgment, relative to tumor staging and the risks from iatrogenic hyperthyroidism. Despite current technical limitations, serum Tg measurement is the cornerstone of long-term monitoring for most DTC patients. For optimal use of serum Tg, it is necessary to understand the pathophysiology of Tg secretion, the limitations of Tg methods and the use of rhTSH to overcome the insensitivity of current Tg methods. PMID- 10365674 TI - Detection of residual and recurrent thyroid cancer by radionuclide imaging. AB - Radioiodine-131 imaging is the traditional method of detecting residual or recurrent differentiated thyroid cancer. The stimulation of such tissues to take up radioiodine may be achieved either by complete cessation of thyroid hormone, by partial thyroid hormone withdrawal, or by the administration of recombinant human thyrotropin (TSH). Complete or partial thyroid hormone withdrawal may result in serum TSH concentrations adequate for radioiodine imaging in up to 90% of patients. When known or suspected recurrent or metastatic disease is not evident on radioiodine imaging, single photon emission tomographic imaging with either thallium-201 chloride or technetium-99m-MIBI compounds may detect up to 80%-90% of cancers at least 1 to 1.5 cm in size, although specificity is less than with 131I. Fluorine-18-FDG positron emission tomography is a somewhat less available but acceptable substitute for thallium-201 or 99mTc-MIBI imaging. Tumor foci that concentrate either TI-201 or 18FDG intensely with little or no 131I uptake appear to behave more aggressively than those concentrating 131I avidly. PMID- 10365675 TI - Development and in vitro characterization of human recombinant thyrotropin. AB - We have gained insight into the molecular mechanism of human thyrotropin (hTSH) action through cloning of the human TSHbeta subunit gene, development of recombinant TSH and novel analogues and chimeras produced by site-directed as well as cassette mutagenesis. A variety of loss of function mutations have shown several key domains in both the alpha- and beta-subunits that are important for high-affinity ligand interaction with the receptor. In contrast the specificity of receptor interaction was shown to be determined primarily by areas within the hTSH-beta "seat-belt" region. We have also designed various gain of function mutants (superagonists) using evolutionary considerations, homology modeling, and sequence comparisons within the cystine knot growth factor superfamily. Such superagonists resulted from increasing the positive charge by introduction of lysine or arginine residues or neutralization of negatively charged residues of the peripheral hairpin loops of each subunit in various combinations. Certain superagonists increased receptor binding, in vitro and in vivo bioactivity 100- to 1000-fold, more than that achieved previously for any other known protein ligand. In vivo metabolic clearance and biologic activity could be separately modulated by alteration of TSH carbohydrate structure including production of chimeras that added sites of O-glycosylation and/or covalently linked the alpha- and beta-subunits. These data suggest that electrostatic interactions resulting from net positive charge in TSH and net negative charge in its receptor play an important role in high-affinity TSH receptor binding and signal transduction. Insights gained from the design of such novel recombinant TSH analogues and chimeras should have many diagnostic and therapeutic applications. These include the design of improved in vitro assays for thyrotropic factors as well as the design of second generation recombinant TSH analogues for the detection and treatment of thyroid cancer. PMID- 10365676 TI - Pharmacology of bovine and human thyrotropin: an historical perspective. AB - Before the induction of a brief period of hypothyroidism became the standard method for inducing 131I uptake in thyroid cancer diagnosis and therapy, several other methods were explored and used. At the dawn of this new era of recombinant human thyrotropin (TSH) it is of interest to reflect briefly on some of this work. Partially purified bovine TSH (bTSH) was supplied for many years by the Armour Company as Thytropar for intramuscular injection and was first used in thyroid cancer 50 years ago at the Montefiore Hospital and at the Memorial Sloan Kettering Cancer Center in New York City. Most of the patients were already hypothyroid and bTSH induced further 131I uptake in only a few. Experience over the next 30 years revealed frequent allergic reactions, occasionally serious ones, and in 1961 it was shown that prolonged use could result in resistance to both bTSH and human TSH. bTSH was, therefore, reserved for thyroid cancer patients unable to increase endogenous TSH when hypothyroid. bTSH also was used widely as a test to distinguish between hypothyroidism caused by thyroid or pituitary failure until it was replaced by thyrotropin-releasing hormone (TRH). In a few studies, TRH was also tested as an adjuvant to increase endogenous TSH and thus help to stimulate function in thyroid cancer, but this attracted little interest. Prolonged hypothyroidism, enhanced by antithyroid drugs, was used early on, but this very effective stimulant of thyroid cancer function was, for multiple reasons, discarded. Beginning interest 15 to 25 years ago in obtaining TSH from human pituitary glands, a byproduct of growth hormone production, was interrupted when this product was found to risk development of Creutzfeld-Jakob disease. Recombinant human TSH, a safe and effective substitute, is now ready for widespread use and development in thyroid cancer management. PMID- 10365677 TI - Analysis of human sodium/iodide symporter, thyroid transcription factor-1, and paired-box-protein-8 gene expression in benign thyroid diseases. AB - The ability to concentrate iodide, a fundamental property of normally functioning thyroid tissue, is altered in various thyroid diseases. Given the critical role of the Na+/I- symporter (NIS) in controlling iodide access to the thyroid gland, altered expression of NIS may be responsible, at least in part, for an enhanced or diminished capacity to concentrate iodide. In this study, we used Northern blot analysis, a newly established quantitative polymerase chain reaction (PCR) assay and in addition hNIS-directed immunohistochemical analysis to assess the levels of hNIS mRNA and protein expression in various localized and diffuse benign thyroid abnormalities, including Graves' disease (GD), scintigraphically cold solitary benign thyroid nodule (CBTN), nontoxic multinodular goiter (NMNG), solitary autonomously functioning thyroid nodule (AFTN), and mild diffuse iodine deficiency goiter (IDG). In addition, in view of the recent identification of putative binding sites for the transcription factors thyroid transcription factor 1 (TTF-1) and human paired-box-protein-8 (Pax-8) in the human NIS gene promoter, we used reverse transcriptase-polymerase chain reaction (RT-PCR) to assess in these same samples the levels of TTF-1 and Pax-8 gene expression. Northern blot analysis revealed high levels of hNIS gene expression in thyroid specimens derived from patients with GD and AFTN. In contrast, levels of hNIS mRNA expression were moderate in NMNG, low in diffuse IDG, and very low in CBTN. Quantitative RT-PCR analysis of hNIS mRNA transcripts revealed variable but generally low levels of hNIS gene expression in IDG and NMNG, and undetectable or very low levels of hNIS mRNA in all scintigraphically CBTN studied. In contrast, markedly elevated levels of hNIS mRNA transcripts were detected in active GD (up to 17-fold) and AFTN (up to 25-fold). Immunohistochemical analysis revealed abundant hNIS protein expression by thyroid follicular cells in GD, moderate and heterogeneous levels in NMNG, and very low levels in CBTN. hNIS mRNA levels were correlated with TTF-1 and Pax-8 gene expression in GD and, to a lesser degree, in AFTN, NMNG, and IDG, but not in CBTN. In general, hNIS gene expression was more closely correlated with TTF-1 as compared to Pax-8 gene expression. In conclusion, the abundance of hNIS mRNA and protein expression in a broad range of benign thyroid pathologies correlated well with their functional state as assessed by thyroid scintigraphy. In addition to TTF-1 and Pax-8, other transcription factors and enhancer elements may contribute to regulation of NIS gene promoter activity. PMID- 10365678 TI - Direct binding of thyrotropin receptor autoantibody to in vitro translated thyrotropin receptor: a comparison to radioreceptor assay and thyroid stimulating bioassay. AB - Graves' disease is characterized by the presence of autoantibodies to the thyrotropin receptor (TSHR), which are pathogenic and responsible for disease activity. It is well recognized that the autoantibodies are heterogeneous and recognize a number of different conformational dependent epitopes on the TSHR. In this study, we have extended our previous observations to study the interaction of Graves' disease autoantibodies with TSHR ectodomain produced by in vitro transcription and translation reaction. The specific activity of the translated TSHR ectodomain has been increased by a log fold by adding an efficient ribosome binding Kozak sequence before the translation initiation codon as well as double labelling with 35S-methionine and 35S-cysteine during the translation reaction. Addition of canine pancreatic microsomes to the translation mix showed that the glycosylation of TSHR ectodomain did not occur efficiently for the nascent receptor protein. In order to determine the specificity and sensitivity of the improved assay with nonglycosylated TSHR ectodomain, we have studied 331 sera from Graves' disease patients and as controls 100 sera from patients with nonthyroid autoimmune disorders as well as sera from 200 normal control subjects with no family history of thyroid autoimmunity. With this large cohort of sera from Graves' disease and control individuals, 25% of Graves' disease sera immunoprecipitated the dual labeled, in vitro transcribed and translated TSHR ectodomain, exceeding the 98th percentile of the control sera. There was no correlation between the autoantibodies that immunoprecipitate the in vitro translated TSHR ectodomain and those that inhibit iodinated TSH binding in the radioreceptor assay and those with biological activity in a bioassay. The data are consistent with the finding that a proportion of Graves' disease autoantibodies can interact directly with TSHR ectodomain produced by in vitro transcription and translation. However, in contrast to the wide use of similar translation and immunoprecipitation assays to measure other autoantibodies for the diagnosis of autoimmune disorders, such as type 1 diabetes, the TSHR immunoprecipitation on its own is unsuitable for diagnosis of Graves' disease. PMID- 10365679 TI - Transition of nodular toxic goiter to autoimmune hyperthyroidism triggered by 131I therapy. AB - The use of 131I treatment in nodular toxic goiter is widely accepted. In this article, we describe transition of nodular toxic goiter into an autoimmune toxic goiter with development of thyrotropin receptor antibodies (TRAb) as a side effect of 131I treatment. In this retrospective study, 149 patients with nodular toxic goiter (100 with multinodular goiter, 49 with a solitary autonomously functioning toxic nodule) were studied. Of these 149 patients 100 became permanently euthryoid after 1 dose of 131I, and due to persistent hyperthyroidism, 32 patients needed 2-5 doses to became euthyroid. After becoming euthyroid, none of these 132 patients had relapse of hyperthyroidism in the follow-up period. Based on evaluation of the thyroid hormone variables, 17 of 149 patients had a distinctly different pattern in the changes in thyroid hormones. They developed an increase in FT4I 3-6 months posttreatment after an initial fall in FT4I. Twelve of these 17 patients were treated with antithyroid drugs before the initial 131I dose. On samples of frozen sera (-20 degrees C) anti-thyroid peroxidase (TPO) and TRAb were followed for 6 months after 131I treatment in these 17 patients. A similar follow-up was done in 20 patients (10 with and 10 without antithyroid drug pretreatment), randomly selected from the patients who did not relapse. In the remaining 112 patients, anti-TPO and TRAb levels were measured only before the 131I treatment. Of the 17 patients with relapse, 6 developed TRAb concomitant with recurrence of hyperthyroidism (4% of the study group). In 5 of the 17 patients TRAb values remained absent throughout the follow up period. The remaining 6 patients had elevated TRAb values before 131I treatment. Among the 132 patients who did not relapse, an additional 7 cases with presence of TRAb were found. A total of 9% of the study group was found to have TRAb before 131I pretreatment. Anti-TPO was found in 20 of 149 patients (13%) before 131I treatment. Complications, either hypothyroidism or TRAb-associated hyperthyroidism, were seen in 8 of 20 patients (40%) with anti-TPO before 131I treatment, compared to 9 of 129 (7%) without (p<0.005). In conclusion, TRAb and a Graves' like hyperthyroidism can be triggered by 131I treatment in patients with nodular toxic goiter. The presence of anti-TPO seem to be a marker of an increased risk of development of TRAb-associated hyperthyroidism as well as hypothyroidism, but both side effects can be seen despite the absence of anti-TPO autoantibodies. PMID- 10365680 TI - Graves-Basedow disease goiter: a model of Bax-Bcl2 regulated apoptosis. AB - This study demonstrates the involvement of a Bax-Bcl2-dependent apoptotic process in Graves-Basedow thyroid disease, a pathological condition known for its spontaneously oscillating evolution. A continuous series of 86 cases of surgically treated Graves' thyroid was evaluated for apoptotic cell content identified by histological criteria and confirmed by terminal desoxynucleotidyl transferase-mediated desoxyuridine triphosphate nick end-labeling (TUNEL). A significant correlation was found between tissue features of Graves' disease (epithelial hyperplasia, cellular hypertrophy, colloid content) and the amount of apoptotic cells. No correlation was found with lymphocytic infiltrates. Significantly, 11 cases (about 12% of the series) with high-level apoptosis displayed the typical features of active Graves' disease over all tissue sections. In contrast, cases with no detectable apoptosis exhibited regressive tissue features of Graves' disease. An intermediate group of cases was characterized by tissue heterogeneity with hyperactive foci, rich in apoptosis, alternating with regressive areas lacking apoptosis. In this group the participation of apoptosis to the remodeling of Graves' thyroid parenchyma, in a tight balance with cell proliferation, was best illustrated. Moreover, the thyroid follicle by accumulating apoptotic cells and bodies, allowed a tentative chronological ordering of apoptosis steps in correlation with Bax-Bcl2 tissue distribution and cellular pattern. Our observations suggest that the initiation of apoptosis corresponds to a loss of cellular cohesion, a drop in Bcl2 expression, and a delocalization of Bax from a putative Golgi storage location to a mitochondrial distribution. PMID- 10365681 TI - Elevated anti-galactosyl antibody titers in endemic goiter. AB - Anti-Gal is a human polyclonal antibody that constitutes approximately 1% of the circulating immunoglobulin G (IgG), interacts specifically with the mammalian carbohydrate alpha-galactosyl epitope. Furthermore, it was found to mimic in vitro thyrotropin (TSH) effects regarding stimulation for cyclic adenosine monophosphate (cAMP) synthesis, 125I uptake, and cellular proliferation on cultured porcine thyrocytes and on Graves' disease thyrocytes, but not on normal human thyrocytes. As immune activation in sporadic and endemic goiters might play a secondary role in regulating thyrocyte proliferation and function, we evaluated anti-Gal titers in endemic goiter. Serum was obtained from 109 Chagas'-negative patients living in an endemic goiter area of Brazil (Grao Mogol, MG) and 160 controls. The patients were divided into 3 groups, according to their goiter size (World Health Organization [WHO] classification): grade 0 (group 1, n = 24), grade I-II (group 2, n = 41), and grade III-IV (group 3, n = 44). Anti-Gal was assessed by a radioimmunological procedure (results expressed as the percentage of bound radioactivity/total activity [%B/T]). The antibody titer was significantly more elevated in group 1 (mean +/- SEM: 9.27%+/-0.80%), in group 2 (mean +/- SEM: 16.17%+/-0.97%), and in group 3 (20.97%+/-1.30%) than in normal controls (6.46%+/-0.33%). Analysis of the male and female data separately for anti-Gal titer did not substantially alter these results. We concluded that the anti-Gal titer is higher in patients with endemic goiter and presented a possible relationship with the size of goiter. Whether these antibodies contribute to the pathogenesis of the disease needs further clarification. PMID- 10365682 TI - Struma ovarii and hyperthyroidism. AB - Struma ovarii is a teratoma of the ovaries that contains a large amount of thyroid tissue. Like the cervical thyroid gland, this ectopic thyroid tissue can become autonomous. We present a case of hyperthyroidism caused by thyroid tissue in a large ovarian cystic teratoma and provide detailed endocrinological, radiological, and pathological preoperative and postoperative data. This is also the first documented case of struma ovarii in association with a secreting pituitary tumor. In addition, we provide a retrospective pathological analysis of 1390 surgically removed ovarian tumors at 2 major academic centers. PMID- 10365683 TI - The effect of thyroid hormone on size of fat depots accounts for most of the changes in leptin mRNA and serum levels in the rat. AB - The physiological consequences and mechanism(s) for thyroid hormone-induced alterations in serum leptin are not known. To address this, leptin expression in rats was evaluated in relationship to food intake, fat mass, and body temperature in rats with pharmacologically altered thyroid status. Total body weight, food intake, and temperature were decreased in hypothyroid rats. Fat weight was decreased in both chronically hypothyroid and hyperthyroid rats (n = 6/group). Serum leptin was linearly correlated with fat weight, epididymal and retroperitoneal fat leptin mRNA concentration, but not total body weight. Serum leptin was decreased in the chronically hyperthyroid rats. When fat weight was used as a covariant, serum leptin was not different between the three groups. Epididymal fat leptin mRNA was higher in euthyroid (n = 7) than in hypothyroid and hyperthyroid rats. Retroperitoneal fat leptin mRNA was not affected by thyroid status. A positive linear relationship between food intake and free triiodothyronine (FT3) index was observed, but not between food intake and serum leptin alone. In a time course study, serum leptin, epididymal fat leptin mRNA content, and fat weight did not change within 24 hours of high-dose triiodothyronine (T3) (n = 6/group), but both temperature and epididymal fat S14 mRNA content rapidly increased. These findings demonstrate that thyroid state influences circulating leptin levels, but primarily does so indirectly through the regulation of fat mass. Leptin does not influence core body temperature across thyroidal state. Finally, thyroid state is more important to regulate food intake, through an as yet undefined mechanism, than are thyroid state-associated changes in serum leptin. PMID- 10365684 TI - Methimazole and propylthiouracil increase cellular thyroid peroxidase activity and thyroid peroxidase mRNA in cultured porcine thyroid follicles. AB - Methimazole (MMI) and propylthiouracil (PTU) are common antithyroid drugs for treating hyperthyroidism because the 2 drugs inhibit thyroid peroxidase (TPO) catalyzed thyroid hormone formation. We studied whether the 2 drugs actually inhibit cellular TPO activity in cultured porcine follicles. Porcine follicles were cultured in the presence of 1 mU/mL thyrotropin (TSH) for 7 days. Then follicles were exposed to MMI or PTU in the presence of 0.1 microM Kl for 2 days. TPO activity was measured in the 100,000 x g-pellet of the thyroid sonicate by the guaiacol oxidation method. Exposure to MMI (1 microM and 10 microM) or PTU (10 microM and 100 microM) for 2 days caused a significant increase in cellular TPO activity; 100 microM MMI inhibited cellular TPO activity. The presence of cyclic adenosine monophosphate (cAMP)-generating system (forskolin) in TSH-free medium increased MMI-mediated TPO activity. Cyclohexamide inhibited MMI-mediated TPO activation, indicating that new protein synthesis is required for increased TPO activity. Reverse transcriptase-polymerase chain reaction (RT-PCR) showed an increase in TPO mRNA by PTU or MMI. In conclusion, MMI and PTU at therapeutic concentrations can increase TPO mRNA and cellular TPO activity, although the 2 drugs inhibit the TPO-H2O2-mediated catalytic reaction. PMID- 10365685 TI - Lack of thyroglobulin production supports the finding of the FRTL-5 cells with a nondiploid karyotype. PMID- 10365686 TI - Optimization of pelvic heating rate distributions with electromagnetic phased arrays. AB - Deep heating of pelvic tumours with electromagnetic phased arrays has recently been reported to improve local tumour control when combined with radiotherapy in a randomized clinical trial despite the fact that rather modest elevations in tumour temperatures were achieved. It is reasonable to surmise that improvements in temperature elevation could lead to even better tumour response rates, motivating studies which attempt to explore the parameter space associated with heating rate delivery in the pelvis. Computational models which are based on detailed three-dimensional patient anatomy are readily available and lend themselves to this type of investigation. In this paper, volume average SAR is optimized in a predefined target volume subject to a maximum allowable volume average SAR outside this zone. Variables under study include the position of the target zone, the number and distribution of radiators and the applicator operating frequency. The results show a clear preference for increasing frequency beyond 100 MHz, which is typically applied clinically, especially as the number of antennae increases. Increasing both the number of antennae per circumferential distance around the patient, as well as the number of independently functioning antenna bands along the patient length, is important in this regard, although improvements were found to be more significant with increasing circumferential antenna density. However, there is considerable site specific variation and cases occur where lower numbers of antennae spread out over multiple longitudinal bands are more advantageous. The results presented here have been normalized relative to an optimized set of antenna array amplitudes and phases operating at 100 MHz which is a common clinical configuration. The intent is to provide some indications of avenues for improving the heating rate distributions achievable with current technology. PMID- 10365687 TI - A two-parameter method for the estimation of ultrasound-induced temperature artifacts. AB - A two-parameter method for the estimation of ultrasound-induced temperature artifacts was evaluated and compared with other commonly applied methods using analytical solutions to the bioheat equation. The two parameters are the exponent of the assumed temperature decay curve after power is turned off and the baseline temperature. These parameters are found by optimizing the fit of the temperature data from 30 to 60s after power is turned off. The artifact is modelled as a point source at the centre of a Gaussian temperature distribution. The blood flow, baseline temperature, and variance of the Gaussian temperature distribution were varied to simulate different clinical situations. Noise was added to the model to investigate the effects of thermometry resolution and sampling intervals. It was found that for artifacts of < 2 degrees C the two-parameter method had errors of less than 0.25 degrees C, whereas other methods generally had greater errors depending on the conduction rate and blood flow rate. The effects of the temperature sampling interval and resolution on the ability of the methods to estimate the artifact were also investigated, and it was found that the two-parameter method was much more sensitive to these parameters than other commonly applied methods. PMID- 10365688 TI - Effects of whole-body hyperthermia on the canine central nervous system. AB - It has been reported that central nervous system (CNS) tissue may be more heat labile than other tissues of the body. However, no definite information has been available on how much heat CNS tissue can tolerate without sustaining damage during whole-body hyperthermia, especially in a chronic stage. In this study, whole-body hyperthermia was induced in dogs by extracorporeal heating of blood, to determine the effects 7 days after hyperthermia on the canine brain and spinal cord. The temperatures of both the brain and the spinal cord were raised to 42.0+/-0.1 degrees C and maintained at that level for 60 min. Seven days later, all of the dogs were sacrificed by transcardial perfusion using 10% formaldehyde phosphate buffer for microscopic examination. The thermal dose resulted in neither microscopic damage to the CNS nor neurological symptoms, as determined by comparison of microscopic and neurological findings with those of dogs whose brain and spinal cord temperatures were maintained at 37.0 degrees C for 60 min. The findings suggest that, for medical purposes, whole-body hyperthermia appears promising for application at a thermal dose of up to 42.0 degrees C for 60 min. PMID- 10365689 TI - The effects of intentional hyperthermia on the Thrombelastograph and the Sonoclot analyser. AB - The effect of whole-body hyperthermia (WBH) on viscoelastic properties of whole blood, as measured by the thrombelastogram (TEG) and Sonoclot analyser, was investigated in 10 patients undergoing WBH-carboplastin therapy for metastatic disease. Blood was taken from an existing central line at baseline (37 degrees C), during warming (39 and 41 degrees C) and cooling (39 and 37 degrees C). Sonoclot and TEG samples were analysed simultaneously at 37 degrees C and at the patient's temperature with a temperature-compensated unit, except at 41 degrees C for the Sonoclot (maximum temperature adjustment of 40 degrees C). TEG measurements included R time (time to initial fibrin formation [mm]), K time (mm) and alpha angle (degrees) (both reflecting fibrinogen-platelet interaction), maximum amplitude (representing qualitative platelet function [mm]) and per cent fibrinolysis at 30 and 60 min. The Sonoclot ACT (SonACT-secs), initial rate of clot formation (%), time to peak amplitude (min) and peak amplitude of the Sonoclot signature (mm) were recorded. Decreased R time of the TEG compared to a marginally elevated baseline was found at all times during warming and cooling (p < 0.05). The K time was decreased at 41 degrees C compared to a normal baseline (p < 0.05). The SonACT was decreased (from an elevated baseline) at all other times, without differences in measures at patient temperature versus 37 degrees C (p < 0.05). The data suggest acceleration of fibrin formation during WBH to 41 degrees C in patients with malignancy. Implications for defining thromboembolic risk require further investigation. PMID- 10365690 TI - Interaction of hyperthermia with Taxol in human MCF-7 breast adenocarcinoma cells. AB - Hyperthermia treatments (43 degrees C, 1 h) were performed on exponentially growing MCF-7 breast adenocarcinoma cells at the beginning, middle, or end of 24 h incubations of the cells in vitro with Taxol (paclitaxel). When the cells were heated at the beginning or middle of the Taxol incubation, the hyperthermia treatment protected against the toxic effect of each of the Taxol concentrations examined (5, 10 and 100 nM). Consistent with earlier studies, Taxol treatment at 37 degrees C resulted in an accumulation of greater than 94% of the cells in G2/M at 24 h. Heating the cells at the middle or end of the Taxol treatment resulted in a similar accumulation. However, heat treatment during the first hour of Taxol exposure resulted in a significantly smaller percentage of cells (approximately 50%) in G2/M. HPLC analysis showed that at 37 degrees C, Taxol uptake into MCF-7 cells approached maximum within 0.25 h and increased only slightly more over the next 11.75 h. The parental Taxol level was markedly lower by 24 h. In contrast, 1 h hyperthermia treatments at the beginning or middle of the Taxol incubation resulted in higher Taxol concentrations at 12 and 24h, and higher intracellular concentrations overall than at 37 degrees C. These results indicate that hyperthermia inhibits Taxol related cell cycle effects and cytotoxicity, in spite of causing higher concentrations of Taxol to be present in heated cells. PMID- 10365691 TI - Stereologic evaluation of the vasculature in a MX1 xenotransplanted tumour model after combinations of treatment with ifosfamide, hyperthermia and irradiation. AB - The vascularization of tumours is a critical parameter of their growing and metastatic behaviour. However, little is known about the morphologic reactions of the microvasculature, especially the capillary bed of tumours and the adjacent tissue. In this study, the vessels in MX1 xenotransplants in athymic nu/nu nude mice were quantified and the angioarchitecture was visualized with the aim of presenting stereologic parameters of vessels based on a morphometric analysis of post mortem tissue blocks which were processed by standard histological procedures. In order to study changes of the microvasculature of MX1 tumours, the xenotransplanted nude mice were treated by different therapeutic regimens. Standardized hyperthermia, ifosfamide and irradiation therapies were applied. Special interest was focused on early changes of capillaries and of the pre- as well as post-terminal vascular bed. The stereologic evaluation of capillaries and larger vessels immediately after the therapy with ifosfamide and hyperthermia shows an increase of the mean capillary sizes. Furthermore, tumour samples after the 5th day of irradiation (5 x 2 Gy) and combinations of irradiation and chemotherapy treatment have been investigated. After 5 days of irradiation, a significant decrease of the vascular density was found. The results presented here clearly show that the timing and the mode of therapy influence the capillary morphology and periterminal vasculature of xenotransplanted MX1 tumours. PMID- 10365693 TI - Reconstruction of the elbow: therapist's commentary. PMID- 10365692 TI - Reconstruction of the elbow: surgeons' perspective. AB - Great progress has been made in surgery of the elbow during the past decade. Better definition of ligamentous anatomy, recognition of previously undescribed instability patterns, and development of ligament reconstruction techniques have helped improve surgery designed to restore stability of the elbow. Surgical release of elbow contractures helps patients with stiffness gain very satisfactory range of motion. New techniques and advances in prostheses have broadened the indications for total elbow arthroplasty, and cases that were previously considered inoperable can now be treated surgically with excellent outcomes. Elbow surgery enters the new millennium having accomplished much, but problems and challenges remain. PMID- 10365694 TI - The radioulnar joints and forearm axis: surgeons' perspective. AB - Forearm injuries are common and can vary in complexity. The fractures are easily diagnosed, but the associated soft tissue injuries may not be as obvious. Treatment results are dependent not only on the fracture healing but on the return of normal relationships about the distal radioulnar joint, the interosseous membrane, and the proximal radioulnar joint. Alteration in any of these elements can result in permanent loss of forearm rotation and stability. Recent anatomic and biomechanical studies have increased our knowledge of these structures and their role in forearm function. The findings from these studies may lead to earlier repair of soft tissue injuries about the radioulnar joints and the interosseous membrane. Further follow-up of these procedures will be needed to validate their indications. PMID- 10365695 TI - The radioulnar joints and forearm axis: therapist's commentary. PMID- 10365696 TI - Management of fractures of the distal radius: surgeon's perspective. AB - The new millennium represents a time for expanding our present knowledge of the treatment of distal radius fractures, based on the foundation of information that has been gathered over the past century. Treatment-oriented classifications have replaced the prior generalization applied to all "Colles fractures" and have directed preoperative planning. Newer external fixation frames, improved surgical technique, and superior instrumentation allow for safer reproducible ligamentotaxis. These current concepts of treatment along with a comprehensive therapy plan have provided the basis for enhanced recovery from these challenging injuries. PMID- 10365697 TI - Management of fractures of the distal radius: therapist's commentary. PMID- 10365698 TI - The carpus: surgeon's perspective. PMID- 10365699 TI - The carpus: therapist's commentary. AB - This paper presents an overview of therapy for the wrist after soft tissue reconstruction, arthroscopic debridement, arthroscopic ganglionectomy, total wrist arthrodesis, and vascularized bone grafts. Postoperative problems common to each of these procedures include stiffness, edema, scarring, decreased strength, and decreased use of the involved extremity. An understanding of the surgical procedure and its specific purpose is necessary for the hand therapist to formulate a plan of treatment and appropriate goals for rehabilitation. Treatment techniques and modalities for these procedures are reviewed, as well as the importance of patient education and activity modification. PMID- 10365700 TI - Bone and joint injury in the hand: surgeon's perspective. PMID- 10365701 TI - Bone and joint injury in the hand: therapist's commentary. PMID- 10365702 TI - Arthroplasty of the hand and wrist: surgeon's perspective. PMID- 10365703 TI - Arthroplasty of the hand and wrist: therapist's commentary. PMID- 10365704 TI - Treatment of peripheral nerve injuries: surgeons' perspective. PMID- 10365706 TI - Treatment of flexor tendon injuries: therapist's commentary. PMID- 10365705 TI - Treatment of flexor tendon injuries: surgeons' perspective. AB - Medical researchers continue to explore the flexor tendon's response to injury and repair. In recent years, hand surgery and therapy publications have focused on the biomechanics of suture techniques and the benefits of early postoperative motion on surgically repaired flexor tendons. Laboratory and clinical studies have shown that stronger suture techniques can withstand the strain of immediate active motion without a significant risk of tendon rupture or gap formation. Newly proposed therapy techniques and anatomic studies defining the effects of wrist and digital position on tendon excursion share the goals of achieving early motion and reducing restrictive adhesions. Clinical studies have evaluated the various imaging modalities used to diagnose postoperative adhesions. Other clinical surveys have detailed the use of pedicled autograft and allograft tendons in staged reconstruction. Histologic and immunologic researchers have concentrated on cellular activation patterns following tendon injury and the effects of pharmacologic agents, such as hyaluronan and aprotinin, on tendon healing and adhesion formation. PMID- 10365708 TI - Vascular disorders of the hand: therapist's commentary. PMID- 10365707 TI - Vascular disorders of the hand: surgeons' perspective. AB - Vascular disorders of the upper extremity present a complex and challenging problem to the treating physician. The presentation is often subtle and the consequences of misdiagnosis or mistreatment can be severe. A thoughtful and through approach combining the history, physical findings, and use of appropriate diagnostic aids will provide the physician and patient with the greatest opportunity for a satisfactory outcome. PMID- 10365709 TI - Treatment of congenital differences of the upper extremity: surgeon's perspective. PMID- 10365710 TI - Treatment of congenital differences of the upper extremity: therapist's commentary. PMID- 10365711 TI - Hand care in the new millennium: surgeon's perspective. PMID- 10365712 TI - Hand care in the new millennium: therapist's commentary. PMID- 10365713 TI - Advocacy group targets PVC i.v. equipment. PMID- 10365714 TI - Breast cancer studies yield mixed results on high-dose chemotherapy with stem cell support. PMID- 10365715 TI - Studies of high-dose chemotherapy with stem cell support for breast cancer. PMID- 10365716 TI - Panel sees medical potential in standardized cannabinoids but not in smoked marijuana. PMID- 10365717 TI - Survey explores consumer views on government regulation of supplements. PMID- 10365718 TI - The need for legal compliance plans in health institutions. PMID- 10365719 TI - Kava: Piper methysticum. PMID- 10365720 TI - Hepatitis C: natural history, diagnosis, and management. AB - The virology, epidemiology, clinical spectrum, diagnosis, and management of hepatitis C are reviewed. Hepatitis C infection is responsible for most cases of chronic viral hepatitis in the United States and is the major reason for liver transplantation. Hepatitis C virus (HCV) is divided into six major genotypes, with type 1 being the most prevalent in the United States. Direct percutaneous exposure is the main route of HCV transmission. Diagnosis of the infection is made by HCV antibody testing or direct detection of HCV RNA in serum. Liver biopsy is recommended for evaluating disease severity before treatment is started. The currently approved treatment for patients with chronic hepatitis C who have elevated liver transaminases and compensated liver disease consists of interferon alfa alone or in combination with ribavirin. Rates of sustained biochemical and virological responses in the range of 20-40% have been reported for a 12-month regimen of interferon alfa. Combination therapy with ribavirin improves these rates. Response rates are lower in patients with HCV genotype 1, a serum HCV RNA concentration higher than 1 million copies/mL, and cirrhosis. In patients who relapse after an initial response to interferon alfa, retreatment with interferon alfa plus ribavirin or with a higher dosage of interferon alfa is recommended. New agents under development for use against hepatitis C include viral enzyme inhibitors, ribozymes and antisense oligonucleotides, and immunomodulators. Further research is needed to optimize existing strategies for treating hepatitis C and to develop new, more effective therapies. Ultimately, combination therapies may hold the key. PMID- 10365721 TI - Effect of pharmaceutical services on adherence to criteria-for-use guidelines in the operating room. AB - Guidelines for the use of neuromuscular blocking agents (NMBAs) and sedatives during two- to four-hour surgical procedures were developed, and the effect of pharmacy presence on adherence to the guidelines was determined. Differences in cost per dose of the NMBAs pancuronium bromide, cisatracurium besylate, and vecuronium bromide were determined. Pancuronium was designated as the first-line agent in the NMBA guidelines, cisatracurium as the second-line agent, and vecuronium as the third-line agent. In the sedative guidelines, lorazepam was the first-line agent, midazolam was the second-line agent, and propofol was the third line agent. Pharmacy presence in the operating room was provided during January 1997. The pharmacist made a preliminary decision about the most appropriate agent and encouraged guideline adherence. Cost and adherence data were compared with data for November 1996 and March 1997. During January, the NMBA guidelines were followed 75% of the time and the sedative guidelines were followed 15% of the time; the corresponding rates for March were 40% and 12%. Compared with November 1996, a saving of $5.61 per case was observed in January in the NMBA category and a saving of $2.77 was observed in March; between January and March, there was an increase of $2.84 per case. Pharmacy presence in the OR was associated with better adherence to criteria-for-use guidelines for NMBAs and sedatives; NMBA cost savings associated with implementation of the guidelines were higher when a pharmacist was present. PMID- 10365722 TI - Stability of sumatriptan succinate in polypropylene syringes. PMID- 10365723 TI - Compatibility and stability of vincristine sulfate, doxorubicin hydrochloride, and etoposide in 0.9% sodium chloride injection. PMID- 10365724 TI - Robinson-Patman Act and the own-use exemption in health systems. PMID- 10365725 TI - Filgrastim for treatment of neutropenia in a neonate. PMID- 10365726 TI - Intravenous magnesium sulfate treatment in a child with status asthmaticus. PMID- 10365727 TI - Controlling myoclonus after high-dosage morphine infusions. PMID- 10365728 TI - Prophylaxis after nonoccupational exposure to HIV. PMID- 10365730 TI - Alternative method for administering proton-pump inhibitors through nasogastric tubes. PMID- 10365731 TI - New drug overview. Celecoxib. PMID- 10365729 TI - Distinct drug-interaction profiles for statins. PMID- 10365732 TI - Journal references. PMID- 10365733 TI - Safety of low-molecular-weight heparin in pregnancy: a systematic review. AB - Unfractionated heparin (UFH) remains the anticoagulant of choice during pregnancy. Low-molecular-weight heparins (LMWH) are an attractive alternative to UFH due to their logistic advantages and their association with a lower incidence of osteoporosis and HIT. We reviewed all published clinical reports concerning the use of LMWH during pregnancy. In addition, participants of an international interest group contributed a cohort of pregnant women treated with LMWH. Pregnancies were divided into two groups; those with and those without maternal comorbid conditions. The number of adverse fetal outcomes and the occurrence of maternal complications were evaluated in the two groups. In the group of women with comorbid conditions (n = 290), 13.4% of the pregnancies were associated with an adverse fetal outcome. In contrast, in the group of women without comorbid conditions (n = 196), 3.1% were associated with an adverse outcome, which is comparable to that seen in the normal population. We conclude that LMWH appear to be a safe alternative to unfractionated heparin as an anticoagulant during pregnancy. PMID- 10365734 TI - Hormone replacement therapy and circulating ICAM-1 in postmenopausal women--a randomised controlled trial. AB - Hormone replacement therapy may reduce the risk of coronary heart disease but underlying mechanism has not been adequately explained. Recent data suggest that intercellular adhesion molecule 1 (ICAM-1) plays a critical role in early stage of atherosclerosis and may serve as a molecular marker for the development of arterial disease. We investigated the effects of oral and transdermal cyclic oestradiol combined with progesterone on plasma concentration of soluble ICAM-1 (sICAM-1). Thirty-seven healthy postmenopausal women were randomly assigned to receive either oral estradiol valerate or transdermal estradiol both combined with micronized progesterone or no hormonal treatment. Plasma sICAM-1 was assayed at baseline and after a 6-month period. Oral but not transdermal estradiol regimen significantly decreased mean value of sICAM-1 compared with no treatment. Differences in sICAM-1 levels between active treatments were significant. There were no significant changes in mean values of fibrinogen between the three groups. Our results show a favorable effect of oral estrogen plus progesterone on a soluble marker of vascular inflammation and may provide plausible explanation for a cardioprotective effect of hormone replacement therapy among healthy postmenopausal women. PMID- 10365735 TI - Factor XIII Val34Leu is a genetic factor involved in the etiology of venous thrombosis. AB - A mutation in the factor XIII gene (FXIII Val34Leu) gene was recently reported to confer protection against myocardial infarction, but its relationship with venous thrombosis is unknown. In addition, a mutation in the 5'-untranslated region of the FXII gene (46 C->T) was identified which is associated with low plasma levels of the protein. Its prevalence in patients with venous thrombosis is also unknown. We investigated the frequency of the FXIII Val34Leu and FXII 46 C->T mutations in 189 patients with deep venous thrombosis and in 187 age-, gender- and race-matched controls. FXIII Val34Leu was detected in 38.6% of the patients and in 41.2% of the controls. Interestingly, homozygosity for the FXIII mutation was found in 1.6% of the patients and in 9.6% of the controls, yielding an odds ratio (OR) for venous thrombosis of 0.16 (95% CI: 0.05-0.5). The OR for heterozygotes was 1.1 (95% CI: 0.7-1.7). The FXII 46 C->T mutation was detected in 46.0% of the patients and in 48.6% of the controls. The OR for heterozygotes was 0.9 (95% CI: 0.6-1.4) and for homozygotes the OR was 0.8 (95% CI: 0.3-1.9). Our data indicate that the FXII 46 C->T mutation is unlikely to be a major risk factor for venous thrombotic disease. In contrast, the homozygous state for FXIII Val34Leu is a strong protective factor against venous thrombosis, which emerges as a novel genetic factor involved in the aetiology of thrombophilia. PMID- 10365736 TI - Increased levels of factor VIII and fibrinogen in patients with venous thrombosis are not caused by acute phase reactions. AB - Factor VIII activity (factor VIII:C) levels > or =150 IU/dl are associated with a 5- to 6-fold increased risk of venous thrombosis compared to levels <100 IU/dl, and fibrinogen levels > or =5.0 g/l increase the thrombosis risk 4-fold. These high levels are present in 25% resp. 3% of the patients with a first episode of venous thrombosis. These findings were based on measurements after the thrombotic event, so the factor VIII and fibrinogen levels in thrombosis patients may have been influenced by acute phase reactions or ongoing inflammatory responses. In the present study we measured plasma C-reactive protein (CRP) as a sensitive marker of an acute phase reaction in 474 thrombosis patients and 474 age- and sex matched healthy controls, that were part of the Leiden Thrombophilia Study (LETS). Mean and median CRP levels were higher in thrombosis patients than in the controls, suggesting inflammation in some patients. CRP affected both factor VIII and fibrinogen levels, in patients and controls alike. After adjustment for the effect of CRP, high factor VIII:C levels still increased the thrombosis risk 6 fold and high fibrinogen levels 4-fold, which is for both very similar to the risk before correction for CRP levels. These results show that although systemic inflammation may be present in some of the patients, elevated levels of factor VIII:C and fibrinogen were in general not caused by acute phase reactions. This further supports a causal relationship between both high factor VIII:C and fibrinogen levels and venous thrombosis. PMID- 10365737 TI - The risk of recurrent venous thromboembolism in carriers and non-carriers of the G1691A allele in the coagulation factor V gene and the G20210A allele in the prothrombin gene. DURAC Trial Study Group. Duration of Anticoagulation. AB - The results concerning the risk of recurrent venous thromboembolism (VTE) in carriers of the G1691A mutation in the coagulation factor V gene are not consistent and this risk in carriers of the G20210A polymorphism in the prothrombin gene has hitherto not been reported. We followed 534 patients for 48 months after their first episode of objectively documented VTE. The prevalence of the G1691A allele in 467 (87.5%) of the patients and in 207 controls was 25.3% and 8.2%, respectively, in heterozygote form and 2.4% and 0.5%, respectively, in homozygote form. The adjusted odds ratio (OR) for the first VTE was 4.4 (95% CI 2.6-7.8). The risk of recurrent VTE in heterozygotes was not statistically different from non-carriers (17.8% vs 17.6%), with 85% power to detect a hazard ratio of 2.35. Homozygotes had a significantly increased risk (p = 0.036) of recurrent VTE. The prevalence of the G20210A allele in 456 patients and 207 controls was 6.1% and 1.4%, respectively. The adjusted OR was 4.6 (95% CI 1.6 19.3) for the first VTE in 28 carriers of this polymorphism. The risk of recurrent VTE for these was not statistically different from non-carriers with an OR of 0.9 (95% CI 0.2-2.9). PMID- 10365738 TI - Factor V Leiden, prothrombin 20210G-->A and the MTHFR C677T mutations in childhood stroke. AB - Ischaemic stroke is a rare occurrence in children and in a proportion of cases the aetiology remains unknown. We have investigated the role of thrombophilia in the aetiology of this condition. Of 50 cases identified at two centres, 37 were available for detailed haematological analysis. No cases were identified with deficiencies of antithrombin, protein C or protein S. One case had elevated IgG anticardiolipin antibodies at low titre. The prevalence of the prothrombin 20210 G-->A mutation, factor V Leiden (FVL) mutation and the C677T mutation in the MTHFR gene was compared in cases to that observed in random unselected cord blood controls. The odds ratio for stroke was not significantly increased in carriers of the prothrombin mutation (OR 1.2; 95% CI 0.1-10.7), FVL (OR 2.5; 95% CI 0.5 13.5), or the C677T mutation (OR 1.7; 95% CI 0.6-4.5). Our findings suggest that thrombophilia may not play a significant role in the aetiology of stroke in children, although a large prospective study is required to investigate this area further. PMID- 10365739 TI - The risk of thrombosis in patients with lupus anticoagulants is predicted by their specific coagulation profile. AB - Lupus anticoagulants belong to the family of antiphospholipid antibodies. They include two phospholipid-dependent inhibitors of coagulation that may be distinguished on the basis of specific coagulation profiles generated from the comparison of the ratios of the Kaolin Clotting Time (KCT) and the dilute Russell's Viper Venom Time (dRVVT): when the ratio of the KCT exceeds that of the dRVVT, the plasma is allocated to the "KCT" coagulation profile, when the opposite occurs, the plasma is defined to belong to the "dRVVT" coagulation profile group. We prospectively followed-up a historical cohort of 100 consecutive patients with lupus anticoagulants referred to our Institution between January 1988 and October 1997 to investigate the relationship between their coagulation profile at diagnosis and the development of thrombosis during a median follow-up time of 37.5 months (range 1-115 months). Fifty-six patients were allocated to the "dRVVT" coagulation profile, whereas the other 44 displayed the "KCT" profile. Lupus anticoagulants were transient in 17 patients, without differences between the two groups. None of these patients developed clinical events before disappearance of the phospholipid-dependent inhibitors of coagulation. The 83 cases with persistent lupus anticoagulants consistently displayed the same coagulation profile they had been allocated to at entry. Fourteen patients developed 18 thromboembolic events during the follow-up, with an overall rate of thrombosis of 4.2% patients-year. Twelve of them belonged to the "dRVVT" coagulation profile, whereas the other 2 to the "KCT" profile (p = 0.03). The "dRVVT" coagulation profile gave an odds ratio of thrombosis of 5.25 (95% confidence interval [C.I]: 1.17-23.50). Ten of the 14 patients who developed thrombosis during follow-up had already experienced thrombosis: a previous thrombotic event caused an odds ratio of recurrency of 2.72 (95% C.I.: 0.85-8.73) (p = 0.09). By multivariate analysis, the "dRVVT" coagulation profile was still associated with a trend to a higher risk of thrombosis, but the difference did not reach statistical significance. Increased levels of anticardiolipin antibodies (> 40 GPL and/or MPL units) were found in all the 14 patients (p = 0.0064). The "KCT" coagulation profile was significantly associated (p = 0.005) with moderate thrombocytopenia (platelets 50-150 X 10(9)/l). Neither profile was found to represent a risk factor for the development of recurrent miscarriages, neoplastic diseases and death. In conclusion, the "dRVVT" profile appears to have predictive value with respect to the thrombotic complications suffered by patients with antiphospholipid antibodies. PMID- 10365740 TI - Predictive value of coagulation markers concerning clinical outcome 90 days after anterior myocardial infarction. AB - To study the predictive value of coagulation markers concerning clinical outcome, prothrombin fragment F1.2 (F1.2), fibrin monomer antigen (FM), D-Dimer (DD), and fibrinogen were measured in plasma samples drawn 2 and 7 days after acute myocardial infarction (AMI) in 314 consecutive patients randomized in a clinical trial of low molecular weight heparin (Dalteparin) (the FRAMI trial). Placebo treated patients suffering death or new AMI within 90 days had significantly higher levels at day 2 of FM (Enzymun-Test FM), and DD (TINAquant D-dimer) (p = 0.001 and 0.02, respectively), but not F1.2 (Enzygnost F1.2 micro), relative to those without serious clinical events. At day 7 all three coagulation activation markers were significantly higher in patients with subsequent adverse clinical outcome. The Dalteparin group had significantly lower levels of these markers as compared to the placebo group. Left ventricular (LV) thrombus formation was not associated with changes in coagulation activation. However, patients with thrombus had significantly higher fibrinogen levels than those without thrombus (p = 0.004 day 2), independent of treatment group. Thus, markers of coagulation activation may be useful in stratification of patients when estimating risk for adverse clinical outcome after AMI. Furthermore, elevated fibrinogen levels are associated with increased risk of LV thrombus formation. PMID- 10365741 TI - Blast cell-surface and plasma soluble urokinase receptor in acute leukemia patients: relationship to classification and response to therapy. AB - Plasminogen activation in leukemia has been less well characterized than in other malignancies. However, the increased tendency to bleeding and tissue infiltration by leukemic cells are processes in which plasminogen activation may be involved. We have examined plasma and the peripheral blood mononuclear cell fraction from 80 patients including 53 patients with newly diagnosed acute leukemia and 27 patients with other hematological disorders as well as 21 healthy controls. In 28 of 29 examined patients with acute myeloid leukemia (AML) and in two of three patients with hybrid leukemia we found urokinase receptor (uPAR) on the cell surface, while most (7/9) samples from patients with acute lymphoblastic leukemia (ALL) were negative for uPAR. The plasma mean value for soluble uPAR (suPAR) was significantly elevated in patients with AML and ALL. In AML the highest values were found in patients who had residual disease after several cycles of chemotherapy. Compared to controls the uPA antigen levels in patient plasmas were elevated and decreased along with uPAR during treatment. Our results suggest that cell surface uPAR may be a useful marker for leukemia classification and in our material a high level of plasma suPAR correlated with resistance to chemotherapy in AML. PMID- 10365742 TI - Clotting activation after transjugular intrahepatic portosystemic stent shunt. AB - BACKGROUND AND AIM: Aim of the study was to investigate the behaviour of clotting system in peripheral circulation of cirrhotic patients undergoing transjugular intrahepatic portosystemic stent shunt (TIPS). METHODS: Clotting variables and endotoxemia were measured 48 h and 30 days after TIPS in patients randomised to receive heparin or not. RESULTS: Forty-eight hours after TIPS, a significant increase of prothrombin fragment F1+2 was observed; such increase was less evident in patients given heparin. Similar findings were observed for endotoxemia, which, however, was not affected by heparin treatment. Thirty days after TIPS procedure prothrombin fragment F1+2 and endotoxemia returned to baseline values independently of the treatment given. CONCLUSION: This study shows that TIPS is associated with an increase of clotting activation which might contribute to acute thrombosis observed after this procedure. PMID- 10365743 TI - Antithrombotic drugs in the primary medical management of intermittent claudication: a meta-analysis. AB - BACKGROUND: There is no consensus on the efficacy of the antithrombotic drugs available for patients with intermittent claudication. METHODS: A Medline and manual search was used to identify relevant publications. Uncontrolled or retrospective studies, double reports or trials without clinical outcomes were excluded. Included studies were graded as level 1 (randomised and double- or assessor-blind), level 2 (open randomised), or level 3 (non-randomised comparative). Mortality, cerebro- or cardiovascular events, amputations, arterial occlusions or number of revascularization procedures performed in the lower limbs, pain-free and total walking distance, ankle brachial index and calf blood flow, were the main outcomes considered. When feasible, end of treatment results, either continuous or binary, were combined with appropriate statistical methods. RESULTS: Mortality was significantly decreased by ticlopidine compared to placebo (common odds ratio 0.68, 95% C.I., 0.49 - 0.95); clopidogrel decreased vascular events in comparison to aspirin (odds ratio 0.76, 95% C.I., 0.63 - 0.92) in level 1 studies. Arterial occlusions and the number of revascularization procedures performed were statistically significantly decreased by aspirin and ticlopidine, respectively. A small but statistically significant improvement in pain-free walking distance was determined by picotamide, indobufen, low molecular weight heparins, sulodexide and defibrotide, in small studies. CONCLUSIONS: Clopidogrel and ticlopidine do reduce clinically important events in patients with intermittent claudication and could be added to the primary medical treatment of these patients. The use of aspirin in these patients cannot be based on direct evidence, but only on analogy with coronary and cerebral atherosclerosis, where it has documented efficacy. Other antithrombotic drugs were not properly evaluated in patients with intermittent claudication. PMID- 10365744 TI - Longitudinal analysis of factor VIII inhibitors in a previously untreated mild haemophilia A patient with an Arg593-->Cys substitution. AB - Recent studies suggest that certain missense mutations associated with mild to moderate haemophilia A predispose to inhibitor development. In this study, we present a longitudinal analysis of the epitope specificity of an inhibitor that developed in a mild haemophiliac with an Arg593-->Cys mutation. Immunoprecipitation studies revealed the presence of antibodies directed towards the light chain and A2 domain of factor VIII. Limited reactivity was observed with metabolically labelled C2 domain. Almost complete inhibitor neutralization was achieved upon addition of A2 domain. Binding of the inhibitor was not affected by the presence of the Arg593-->Cys substitution in the recombinant A2 fragment. Evaluation of the epitope specificity of anti-factor VIII antibodies in plasma samples obtained at different time-points of inhibitor development revealed initial development of a low titre inhibitor directed towards the A2 domain and factor VIII light chain. A second period of factor VIII replacement therapy resulted in a dramatic rise in factor VIII inhibitor titre, which maintained their original epitope specificity. Based on the results of this and previous studies (Fijnvandraat et al., 1997; Thompson et al., 1997) it is argued that inhibitor development in patients with the Arg593-->Cys mutation may proceed via a similar mechanism. PMID- 10365745 TI - Production of recombinant human protein C in vitro and in vivo by muscle cells. AB - Protein C plays a key role in a natural anticoagulation mechanism, and is also implicated in fibrinolytic and anti-inflammatory functions. Here we describe the production of biologically active human protein C by muscle-targeted gene transfer. Human protein C expression vectors were designed and constructed to produce human protein C in skeletal muscle cells. These vectors were tested in transient and stable transfections of SCID mice myoblasts. Stably transfected cells produced as high as 2.27 microg/10(6) cells/day. Human protein C produced had a relative activity of 92+/-8% compared to the plasma derived human protein C, and was composed of alpha and beta forms, 69% and 31%, respectively. After implantation of stably transfected myoblasts into the hind limb muscles of SCID mice, systemic stable production of human protein C in a range of 31-116 ng/ml serum was obtained up to at least 2.5 months. PMID- 10365746 TI - Prevalence of prothrombin G20210A, factor V G1691A (Leiden), and methylenetetrahydrofolate reductase (MTHFR) C677T in seven different populations determined by multiplex allele-specific PCR. AB - Individuals belonging to six racial groups (African American, Asian Indian, Caucasian, Hispanic, Korean, Native American), and a seventh group comprised of referred patients with thrombosis were genotyped for the prothrombin G20210A mutation, the factor V G1691A (Leiden) mutation, and the methylenetetrahydrofolate reductase (MTHFR) C677T mutation by multiplexed allele specific PCR. The prothrombin 20210A and factor V 1691A allele frequencies in the thrombosis patients, 3.2% and 9.5%, were significantly higher than those in the random Caucasians, 1.3% and 1.8%, (p = 0.043 and p <0.001, respectively). The relative risk of venous thrombosis was determined to be 2.4-fold for carriers of the prothrombin 20210A allele (odds ratio = 2.54; 95% confidence interval = 0.94, 6.82) and 4.5-fold for carriers of the factor V 1691A allele (odds ratio = 5.06; 95% confidence interval = 2.25, 11.36). Among the seven populations, significant differences were observed in the MTHFR 677T allele distribution, however this mutation was not determined to be a risk factor for venous thrombosis in the patient group studied, either alone or in combination with the prothrombin 20210A and/or the factor V 1691A allele(s). PMID- 10365747 TI - The relationship of mutations in the MTHFR, prothrombin, and PAI-1 genes to plasma levels of homocysteine, prothrombin, and PAI-1 in children and adults. AB - Studies in adults have demonstrated that the genetic mutations C677T methylenetetrahydrofolate reductase (MTHFR), prothrombin 20210A, and the 4G polymorphism of the plasminogen activator inhibitor-1 (PAI-1) gene are associated with elevated plasma levels of homocysteine. prothrombin and PAI-1, respectively and with an increased risk of thrombosis. No similar data is available in children. Therefore, we assessed the relationship of plasma levels of homocysteine, prothrombin and PAI-1 with their respective mutations in 197 normal children, compared to 40 adults. By stepwise multiple regression, homocysteine was positively associated with age, PAI-1 activity was negatively associated with age, while PAI-1 antigen and prothrombin levels were associated with gender, being higher in girls than boys. When the genotypes were added to the regression model as additional explanatory variables, the MTHFR genotype accounted for 2.9% of the variance of homocysteine (p = 0.024), and the PAI-1 gene accounted for 2.7% of the variance of PAI-1 antigen levels (p = 0.023). Of children homozygous for the MTHFR mutation, 35% had homocysteine levels > or = the age-specific 95th percentile, compared to 2% heterozygotes and 5% wild type normals (p = 0.0001). The mean homocysteine level was higher in children homozygous for the MTHFR gene (8.4 micromol/1) than in heterozygotes (5.5 micromol/l), p <0.05. Of children homozygous for the 4G polymorphism of the PAI-1 gene, 19% had PAI-1 activity levels > or = the age-specific 95th percentile, compared to 2% of heterozygotes and 3% of wild type normals (p = 0.003). Studies of the incidence of the MTHFR, prothrombin, and PAI-1 4G/5G genotypes in children with thrombosis, when compared to these healthy normals, will provide evidence as to which of these genes are associated with thrombophilia. PMID- 10365748 TI - FXII (46C-->T) polymorphism and in vivo generation of FXII activity--gene frequencies and relationship in patients with coronary artery disease. AB - Increased Factor XIIa concentrations have been found in association with coronary artery disease. Recently, a common 46 C to T point mutation in exon I of the factor XII gene has been described which is associated with lower FXII clotting activity and lower zymogen levels in relation to possession of the T allele. It is not known whether this polymorphism relates to the phenotypes of FXIIa in vivo or to coronary artery disease. The aim of the study was to investigate the interaction of this polymorphism with FXIIa plasma levels and to study the prevalence of the polymorphism in 266 patients with suspected coronary artery disease characterised by angiography and in 185 healthy controls. FXIIa levels were strongly associated with FXII genotype with lower levels with increasing numbers of T alleles (p <0.0001). There was no difference between the prevalence of this polymorphism in patients with M1 compared to those without MI and controls and between all patients and controls (p > or =0.2, chi-square test). There was no association between extent of coronary artery disease (0, 1, 2, and 3 vessel disease) and FXII genotype. In conclusion, the common 46 C to T point mutation is strongly associated with FXIIa but the present study did not show an association with coronary artery disease. The role of this polymorphism in other thrombotic disorders such as ischemic stroke and venous thrombosis and its clinical significance in FXII deficient states remains to be investigated. PMID- 10365749 TI - Enhancement of protein S anticoagulant function by beta2-glycoprotein I, a major target antigen of antiphospholipid antibodies: beta2-glycoprotein I interferes with binding of protein S to its plasma inhibitor, C4b-binding protein. AB - Thrombosis in the antiphospholipid syndrome has been associated with acquired deficiency of the anticoagulant protein S. We sought evidence that beta2 glycoprotein I, a major target antigen for antiphospholipid antibodies, is involved in regulation of protein S activity. Incubation of purified protein S or plasma with beta2-glycoprotein I reversed functional modulation of protein S by its plasma inhibitor, the C4b-binding protein. In a plasma-free ELISA, beta2 glycoprotein I prevented the binding of protein S and C4b-binding protein when preincubated with immobilized protein S but not when similarly preincubated with C4b-binding protein. beta2-glycoprotein I in fluid phase interfered with precipitation of protein S by sepharose-bound C4b-binding protein. Effects of beta2-glycoprotein I on protein S function were inhibited by one of four monoclonal anti-beta2-glycoprotein 1 antibodies. These data suggest that beta2 glycoprotein I helps maintain adequate plasma levels of circulating free, active protein S. Antiphospholipid (anti-beta2-glycoprotein I) antibodies might cause sporadic thrombosis, at least in part, by impairing this novel regulatory mechanism. PMID- 10365750 TI - The sensitivity and specificity of commercial reagents for the detection of lupus anticoagulant show marked differences in performance between photo-optical and mechanical coagulometers. AB - This study was undertaken to appraise the application of those reagents most widely used in the UK for the detection and confirmation of lupus anticoagulant (LA) on an Amelung KC4A and a Sysmex CA-6000 coagulometer. Five sets of dilute Russell's viper venom time (DRVVT) reagents were assessed as well as the Textarin PL/ Ecarin ratio. Each DRVVT method comprised both LA detection and confirmation reagents provided by the same manufacturer. Samples were obtained from 20 normal healthy subjects, 10 LA-positive patients, 10 patients receiving oral anticoagulant therapy (OAT) who had previously been documented as LA-positive, a further 10 LA-negative patients receiving OAT and 10 LA-negative patients receiving unfractionated heparin therapy. The sensitivity and specificity of the reagents exhibited considerable variation not only between reagents, but also when the same reagent was used on the two analysers. Sensitivity ranged from 62 to 97% (all reagents both analysers), specificity went as low as 23% (Gradipore reagent on the CA-6000) and as high as 100% (American Diagnostica Inc on both KC4A and CA-6000). On the KC4A instrument, Unicorn Diagnostics' lupus anticoagulant kit offered the best compromise of sensitivity and specificity (sensitivity 83% and specificity 81%). On the CA-6000 the reagents supplied by American Diagnostica Inc exhibited optimal performance (sensitivity 90% and specificity 100%). The results indicate a need to optimise test reagents for specific analyser types, a procedure which can only be undertaken with preparations such as the proposed NIBSC reference plasmas for the detection of lupus anticoagulant. PMID- 10365751 TI - Fibrinogen Matsumoto III: a variant with gamma275 Arg-->Cys (CGC-->TGC)- comparison of fibrin polymerization properties with those of Matsumoto I (gamma364 Asp-->His) and Matsumoto II (gamma308 Asn-->Lys). AB - Fibrinogen Matsumoto III (M-III) is a dysfibrinogen identified in a 66-year-old woman with rectal cancer. The fibrinogen level determined by the thrombin-time method was markedly decreased in preoperative coagulation tests of her plasma. Three fibrinogen polypeptide-chain gene fragments from the proposita were amplified by the polymerase chain reaction method, then sequenced. The triplet CGC encoding the amino acid residue gamma275 was replaced by TGC, resulting in the substitution of Arg->Cys. There have been previous reports of nine families with the same alteration, nine families with an Arg->His variant and one family with an Arg->Ser variant in this residue, which has been shown to be one of the most important amino acids in the 'D:D' interaction site. In addition, there are three silent mutations in the Aalpha-chain gene and two mutations in the intron of the Bbeta-chain and the gamma-chain gene. However, none of these mutations is thought to be the cause of the dysfunctional fibrinogen. The thrombin-catalyzed fibrin polymerization in the presence of 1 mM Ca ions was markedly delayed in purified M-III. Its lag period was longer than those of Matsumoto II (M-II; gamma308Asn->Lys) and Matsumoto I (M-I; gamma364Asp-His). gamma364Asp is one of the most important residues in the polymerization pocket of the 'D:E' interaction site and gamma308Asn is located in the vicinity of a high affinity Ca2+ binding site in the D-domain, gamma311-336. The maximum slope of the polymerization curve for M-III was about 4-fold steeper than that for M-1 but less steep than that for M-II. These results may suggest that the tertiary structure of the polymerization pocket plays a more important role in the lateral aggregation of protofibrils than that of the 'D:D' interaction site. PMID- 10365752 TI - Transcriptional regulation of the murine urokinase-type plasminogen activator gene in skeletal myoblasts. AB - We have previously shown that urokinase-type plasminogen activator (uPA) is highly expressed in murine C2C12 myoblasts and that antibodies against uPA are able to block both myoblast fusion and differentiation. Here we show the characterization of cis-acting elements in the mouse uPA promoter in vitro which are involved in uPA gene expression in C2C 12 myoblast cells. DNase I hypersensitive (HS) site analysis revealed the presence of three HS sites in myoblasts. Deletion analysis of stably transfected uPA-promoter constructs revealed that at least two of the three HS sites accounted for the high transcriptional expression in C2C12 cells. One was located at -2.4 kb and corresponded to a known PEA3/AP1A element and the other one was located at -4.9 kb and contained a CArG box and a CRE element. So far, no regulatory function had been assigned to this CRE/CArG element. Both HS sites alone were able to activate transcription of a heterologous promoter and showed a cooperative effect when placed together. Electrophoretic mobility-shift assays using myoblast nuclear extracts and specific antibodies demonstrated that cJun, JunD and ATF2 bound to the PEA3/AP1A element, whereas the CRE/CArG element bound SRF. Altogether, these results suggest that high uPA expression in myoblasts is dependent on the cooperation of two regulatory sites in the uPA promoter. PMID- 10365753 TI - The P2Y1 receptor is normal in a patient presenting a severe deficiency of ADP induced platelet aggregation. AB - ADP is a key stimulus inducing platelet shape change and aggregation, a rise in internal calcium and inhibition of adenylyl cyclase. These signaling pathways are thought to be activated by three independent receptors, but to date only the P2Y1 receptor responsible for calcium mobilization and the ionotropic P2X1 receptor have been identified. We report here the characteristics of the P2Y1 receptor in a patient presenting a selective deficiency of ADP-induced aggregation. Cloning of the P2Y1 gene revealed that the patient's DNA and mRNA were normal. Pharmacological studies showed that the P2Y1 receptor was expressed and functional in patient's platelets. Hence, the P2Y, receptor is not the cause of the impaired ADP-induced platelet aggregation in this patient. The P2X1 mRNA was also found to be present and normal. These findings add evidence to previous observations suggesting that a third P2 receptor coupled to adenylyl cyclase may be involved in ADP-induced platelet aggregation. PMID- 10365754 TI - Function of glycoprotein VI and integrin alpha2beta1 in the procoagulant response of single, collagen-adherent platelets. AB - Various collagen-based materials were used to assess the structural requirements of collagen for inducing the procoagulant response of adhering platelets, as well as the collagen receptors involved. Cross-linked or monomeric collagen-related peptide (CRP), Gly-Cys-Hyp-(Gly-Pro-Hyp)10-Gly-Cys-Hyp-Gly was highly adhesive for platelets in a glycoprotein VI-(GpVI-)dependent manner. Adhesion was followed by a prolonged increase in cytosolic [Ca2+]i, formation of membrane blebs, exposure of phosphatidylserine (PS) and generation of prothrombinase-stimulating activity. Fibrils of type-I collagen were less adhesive but, once adhered, many of the platelets presented a full procoagulant response. Monomeric type-I collagen was unable to support adhesion, unless Mg(2+)-dependent integrin alpha2beta1 interactions were facilitated by omission of Ca2+ ions. With all surfaces, however, post-addition of CaCl2 to adhering platelets resulted in a potent Ca(2+)-influx signal, followed by PS exposure and bleb formation. The procoagulant response elicited by binding to CRP was inhibited by anti-GpVI Fab fragments, but not by impeding integrin alpha2beta1-mediated events. With fibrillar collagen, it was inhibited by blocking either the GpVI- or integrin alpha2beta1-mediated interactions. This suggests that the triple-helical Gly-Pro Hyp repeat in CRP and analogous sequences in fibrillar collagen stimulate the procoagulant response of adherent platelets by acting as ligands for GpVI. Influx of Ca2+ is required for this response, and adhesion via integrin alpha2beta1 serves to potentiate the signaling effects of GpVI. PMID- 10365755 TI - Identification of the binding site for an alloantibody to von Willebrand factor which inhibits binding to glycoprotein Ib within the amino-terminal region flanking the A1 domain. AB - An alloantibody to von Willebrand factor (vWF) which developed in a Japanese boy with type 3 von Willebrand disease has been characterized. The antibody was non precipitating IgG and the main subclasses were IgG2 and IgG4. The antibody inhibited completely ristocetin-induced platelet aggregation (RIPA) and high shear stress-induced platelet aggregation (SIPA). Its predominant inhibitory role was focused, therefore, on the interaction between vWF and platelet gycoprotein Ib. The antibody reacted with a 52/48 kDa tryptic fragment of vWF (residues 449 728). No reaction was seen, however, with either a 39/34 kDa dispase fragment (480-718) or a recombinant vWF fragment (residues 465-728). These findings suggested that the essential epitope resided in the amino-terminal flanking region of the Al domain. We synthesized overlapping peptides corresponding to the region containing D3/A1 boundary. A peptide, residues 458-472, bound to the antibody and dose-dependently blocked the antibody binding to the 52/48 kDa fragment. The same peptide neutralized the inhibitory effect of the alloantibody on SIPA. The data are consistent with the presence of an epitope within residues 458-472 which reacted with the 52/48 kDa fragment. Furthermore, the specific component of the antibody, directed against residues 458-472, blocked vWF binding to GPIb in absence of exogenous agonist. Our results suggest that the region flanking the Al domain plays an important role in regulating vWF binding to GPIb. PMID- 10365756 TI - Temporal and topographic matrix metalloproteinase expression after vascular injury in mice. AB - Temporal and topographic expression of matrix metalloproteinases (MMPs) after perivascular electric injury was studied in wild-type (WT) and urokinase deficient (u-PA-/-) mice. Neointima formation after injury of the femoral artery was significantly reduced in u-PA-/- mice as compared to WT mice (area of 0.002+/ 0.0007 mm2 versus 0.008 + 0.002 mm2 at 3 weeks after injury; p <0.001), associated with impaired cellular migration (nuclear cell counts of 44+/-5 versus 82+/-9in cross-sectional areas; p <0.001). Zymographic and/or microscopic analysis indicated that MMP expression gradually increased to reach a maximum at 1 to 2 weeks after vascular injury. In general, MMP levels were lower in u-PA-/- than in WT mice. In non-injured arteries, MMP-2 (gelatinase A) and MMP-3 (stromelysin-1) were produced mainly by adventitial fibroblasts and/or non contractile smooth muscle cells (SMC). One week after injury, MMP-2 and MMP-3 levels were enhanced due to an increased number and size of producing cells; 2 to 3 weeks after injury, MMP-2 and MMP-3 were produced also by some contractile SMC, which stained with alpha-actin antiserum. MMP-9 (gelatinase B), MMP-12 (metalloelastase) and MMP-13 (collagenase-3) were found in macrophages located mainly in the adventitia. Immunogold electron microscopic examination revealed that MMP-2 was located predominantly in association with the cell surface of fibroblasts or SMC, while MMP-9 and MMP- 12 were located in well defined storage granules within macrophages. MMP-2, MMP-3 and MMP-13, but not MMP-9 or MMP-12, were also found extracellularly, associated with elastin-containing structures (MMP-2), with the basement membrane and occasionally with collagen fibres (MMP 3), or with proteoglycans, collagen and elastin (MMP-13). The temporal and topographic expression pattern of MMPs after vascular injury, coinciding with smooth muscle cell migration and neointima formation, thus is compatible with a role in vascular remodeling. PMID- 10365757 TI - Increased expression of protease activated receptor-2 (PAR-2) in balloon-injured rat carotid artery. AB - Protease-induced cell signaling is mediated by specific receptors such as the emerging family of protease activated receptors (PARs). Since proteases are involved in various aspects of vascular injury, we assessed expression of PAR-2, a protease-activated receptor closely related to the thrombin receptor (PAR-1) but activated by an unknown protease, in vascular injury. Rat carotids were subjected to balloon-catheter injury and perfusion fixed at 1, 3, 7 or 14 days after injury. Sections of injured and normal carotid arteries were immunohistochemically labeled with a polyclonal antibody raised against the N terminal residues 37-53 of human PAR-2. Sections were also labeled with antibodies to factor VIII-related antigen, smooth muscle actin and a proliferating cell nuclear antigen (PCNA). In normal vessels, PAR-2 labeling was diffuse and patchy in medial smooth muscle and endothelium. At one and three days after injury, before appearance of neointima, PAR-2 labeling increased in cells adjacent to damaged or necrotic smooth muscle cells. In addition, proliferating adventitial myofibroblasts labeled strongly for PAR-2. At 7 and 14 days after injury, the media and neointima of injured vessels had increased PAR-2 labeling which was most intense at the luminal edge of the neointima. Double immunohistochemical labeling confirmed the greatest expression of PAR-2 in areas with the greatest density of PCNA-positive cells. In addition, PAR-2 mRNA localization using in situ hybridization paralleled PAR-2 expression. The data suggest an upregulation of PAR-2 in response to vascular injury which is associated with medial smooth muscle damage, proliferating adventitial myofibroblasts and smooth muscle cells of the neointima, particularly those at the proliferating luminal edge of the neointima. Possible functional consequences of this receptor upregulation and its role in the response to vascular injury remain to be determined. PMID- 10365758 TI - Plasmodium falciparum-infected erythrocytes: a mutational analysis of cytoadherence via murine thrombomodulin. AB - The pathophysiologic events leading to organ damage in Plasmodium falciparum malaria infections involve adhesion and sequestration of parasite-infected erythrocytes (PRBC) to the vascular endothelium and syncytiotrophoblast. Several potential receptors to which the PRBCs may bind have recently been identified, one of which is thrombomodulin (TM). TM has been implicated particularly in mediating sequestration of P. falciparum-infected erythrocytes in the placenta and brain, two sites of disease associated with high morbidity. In order to establish that binding of parasite-infected red blood cells to TM is dependent on its containing chondroitin-4-sulfate (CSA), we have mutated the CSA-attachment site of murine TM, and expressed this mutant form (TMsergly) in COS-7 cells. In cytoadhesion assays, we demonstrate that, in contrast to wild-type TM which contains CSA and supports the adhesion of 1466 PRBCs/mm2, TMser-gly does not contain CSA and adhesion of PRBCs to those cells expressing TMser-gly is entirely abrogated (200 PRBCs/mm2). These studies further confirm that the CSA of TM may play a role in the pathophysiology of malaria by providing a binding site for PRBCs. PMID- 10365759 TI - Effects of statins in thrombosis and aortic lesion development in a dyslipemic rabbit model. AB - HMG-CoA reductase inhibitors (statins) are effective in primary and secondary prevention of coronary heart disease. The mechanism of action is mainly attributed to their plasma cholesterol lowering activity, although additional effects have been suggested. Our objective was to study whether atorvastatin and simvastatin exhibited an inhibitory effect on platelet deposition onto a triggering damaged vessel wall in addition to an antiatherosclerotic effect in the dyslipemic rabbit model. Statins were administered at identical doses of 2.5 mg/kg/day with a hyperlipidemic diet during 10 weeks. Both drugs similarly lowered total cholesterol and, moderately, triglycerides. Mural platelet deposition on damaged vessel wall placed in an ex-vivo flow perfusion system was reduced in atorvastatin treated animals (39.7+/-6.2 X 10(6) PLT/cm2) vs. controls (94.8+/-15.9 x 10(6) PLT/cm2, p <0.02). Simvastatin reduced aortic fatty streak surface coverage (31,7+/-5.3%) vs. controls (47.9+/-4.1%, p <0.005) and intimal thickening in thoracic aorta (0.15+/-0.05 intima to total area ratio in simvastatin treated animals vs. 0.36+/-0.03 in control animals, p <0.05). Atherosclerotic fatty streak coverage correlated positively with total cholesterol, tryglicerides and LDL-cholesterol levels in all groups. HMG-CoA reductase inhibitors similarly lowered plasma lipids but exhibited significantly different effects in the modulation of atherosclerotic development and platelet response at the tested dose. Therefore, the effect of statins on the progression and manifestation of cardiovascular disease might be also mediated by regulating platelet response to vessel injury. PMID- 10365760 TI - DX-9065a, an orally active factor Xa inhibitor, does not facilitate haemorrhage induced by tail transection or gastric ulcer at the effective doses in rat thrombosis model. AB - DX-9065a is an antithrombin III (AT III)-independent and selective inhibitor of activated blood coagulation factor X (FXa). We evaluated the effects of DX-9065a and warfarin on bleeding time and blood loss in rat tail transection model and on blood loss in hydrochloride (HCl)-induced rat gastrointestinal haemorrhage model. The blood loss was determined by measuring the haemoglobin content in saline immersed with transected tail or hematin chloride content in the gaster after HCl administration. DX-9065a or warfarin was administered orally at 1 h or 15-21 h before the haemorrhagic stimuli, respectively. The dose required for 50% inhibition of thrombus formation (ID50) was 21 mg/kg for DX-9065a and 0.75 mg/kg for warfarin in a copper wire-inserted arteriovenous (AV) shunt model. In contrast to DX-9065a (10 or 30 mg/kg), warfarin (0.75 mg/kg) significantly prolonged the bleeding time. In rat tail transection model, the blood loss for the control group was 102+/-41 microl at 20 min after the transection. While warfarin (0.75 mg/kg) facilitated the blood loss about 5 times as much as the control, DX-9065a (10 or 30 mg/kg) did not. In rat gastrointestinal model, the blood loss for the control group was 15.9+/-5.6 microl at 15 min after HCl administration. In contrast to DX-9065a (10 or 30 mg/kg), warfarin (0.75 mg/kg) increased the blood loss about twice as much as the control. Thus, compared with warfarin, DX-9065a only increased bleeding time or blood loss to a minor extent in the doses tested. These observations suggest that direct inhibition of FXa could be preferable to warfarin in the suppression of thrombosis without haemorrhagic complications. PMID- 10365761 TI - The utility of animal models in the preclinical study of interventions to prevent human coronary artery restenosis: analysis and recommendations. On behalf of the Subcommittee on Animal, Cellular and Molecular Models of Thrombosis and Haemostasis of the Scientific and Standardization Committee of the International Society on Thrombosis and Haemostasis. AB - Small animal models have several advantageous characteristics, but those used in preclinical restenosis research have lacked efficacy in predicting the success of interventions to inhibit restenosis in humans. Large animal models have been more successful than small animal models in predicting efficacy of interventions to inhibit restenosis in humans, but the results of studies carried out with these models have not been uniformly predictive. Confirmation of the results of small animal studies in large animals has not always yielded information predictive of success in humans; however, the absence of such confirmation has had strong negative predictive value. Small animal models used for evaluation of interventions to inhibit luminal narrowing following arterial instrumentation have failed to closely simulate human atherosclerosis and the stenotic lesions subjected to instrumentation in humans. Transgenic, atherosclerotic animals hold promise for the development of more useful small animal models to study mechanisms of the response of diseased arteries to angioplasty and stents. The pig has been the most useful large animal to study stenosis/ restenosis, but more information is needed to overcome the limitations of this model. PMID- 10365762 TI - Placental genotyping of the factor V Leiden, prothrombin 20210A and the methylenetetrahydrofolate reductase (MTHFR) C677T alleles in IUGR pregnancies. PMID- 10365763 TI - Oral contraceptive users and screening of factor V Leiden. PMID- 10365765 TI - Lupus anticoagulant: influence on the international normalized ratio. PMID- 10365764 TI - Can low level of von Willebrand factor decrease the risk of thrombosis in families with antithrombin or protein C deficiency? PMID- 10365766 TI - Most apyrase preparations are impure and contain inhibitors of cathepsin G: suggestions for use of apyrase in preparation and stabilization of platelet suspensions. PMID- 10365767 TI - Circulating vascular endothelial growth factor correlates with systemic thrombin generation in patients with pancreatic carcinoma. PMID- 10365768 TI - Is a change of factor VIII product a risk factor for the development of a factor VIII inhibitor? PMID- 10365769 TI - Endothelial dysfunction in the aorta of transgenic rats harboring the mouse Ren-2 gene. AB - The renin-angiotensin system plays an important role in the pathophysiology of hypertension. We studied vascular function in the aorta of mouse Ren-2 transgenic rats (TGR(mRen2)27). Changes in isometric tension of isolated aorta of TGR(mRen2)27 and Sprague-Dawley rats (SD) were recorded in organ chambers. Contractions to angiotensin II (AII), big-endothelin and endothelin-1 (ET-1), but not KCl were decreased in TGR. Blockade of nitric oxide (NO)-synthase by L-NAME or removal of the endothelium did not alter these decreased contractions to ET-1 and AII in TGR, suggesting that receptors or signaling pathways of these two agonists are downregulated during hypertension. Contractions to norepinephrine (NE) were also lower in TGR, however blockade of NO-synthase by L-NAME or removal of the endothelium evoked similar contractions to NE in both strains, suggesting that basal release of NO reduces contractions to NE to a greater extent in transgenic than control rats. In the presence of L-NAME, acetylcholine evoked endothelium-dependent contractions (EDCF) in TGR, which were blocked by the thromboxane/prostaglandin H2 receptor antagonists SQ 30741, and partially by the thromboxane synthase inhibitor CGS 13080, suggesting that prostaglandin H2 is the mediator. Endothelium-dependent relaxation to acetylcholine was decreased in TGR, while endothelium-independent relaxations to sodium nitroprusside were similar in both strains. SQ 30741 did not improve relaxations to acetylcholine in TGR indicating that impaired relaxations to acetylcholine are due to a decreased acetylcholine-receptor mediated release of NO rather than increased release of EDCF. Thus, Ren-2 hypertension leads to marked alterations of vascular functions in the aorta. These changes could contribute to hypertension and its vascular complications in TGR(mRen2)27 rats. PMID- 10365770 TI - FGF-2 dependent angiogenesis is a latent phenotype in basic fibroblast growth factor transgenic mice. AB - Overexpression of basic fibroblast growth factor (FGF-2) in transgenic (TgFGF2) mice results in a chondrodysplasia as the principle phenotype. Here we report a second phenotype in TgFGF2 mice that was previously undetected: A predisposition to angiogenic reactions with subsequent amplified angiogenesis that are both FGF 2 dependent. We used subcutaneous injection of extracellular matrix as an angiogenic assay. The matrix formed vascularized cysts in the TgFGF2 group after seven days, whereas the non-transgenic (NTg) group developed avascular cysts. Cysts from the TgFGF2 group contained 3-7X more hemoglobin (Hb), two-fold more vonWillebrand Factor (VWF), and 150X more FGF-2 than cysts from the NTg control group. Significant angiogenic reactions occurred only in the TgFGF2 group that express FGF-2 from the transgene. The TgFGF2 mice, therefore, constitute a unique experimental system to study FGF-2 dependent angiogenesis because they have no spontaneous or inherent vascular defects, but provision of an angiogenic substrate results in an amplified angiogenic response. In addition, we report development of an ELISA for VWF that provides a sensitive, quantitative assay for angiogenesis. PMID- 10365771 TI - Ultrastructural localization of nitric oxide synthase and endothelin in the renal and mesenteric arteries of the golden hamster: differences during and after arousal from hibernation. AB - This is a study of the electron-immunocytochemical localization of nitric oxide synthase (type III) and endothelin in renal and mesenteric artery endothelial cells of normal (active) and hibernating hamsters, as well as hamsters exposed to the cold but not hibernating, and hamsters aroused for 2h following hibernation. In the renal artery of hibernating hamsters and cold-exposed hamsters, a subpopulation of nitric oxide synthase-positive endothelial cells displayed immunoprecipitate predominantly in the vicinity of the Golgi complex indicating intracellular translocation from the cytoplasm to the Golgi complex. In hibernating animals, the percentages of both nitric oxide synthase-positive and endothelin-positive endothelial cells were notably lower than those observed either in active, cold-exposed or aroused animals. These changes may reflect a reduced endothelial contribution to the maintenance of vascular tone in these vessels during hibernation and an upregulation of expression of nitric oxide synthase and endothelin in the endothelium early on during arousal from hibernation. PMID- 10365772 TI - Are Angiotensin Converting Enzyme and von Willebrand factor circulating levels useful surrogate parameters to monitor disease activity in Kawasaki disease? AB - To verify if Angiotensin Converting Enzyme (ACE) and von Willebrand factor (vWF) may be used as a laboratory marker for the follow-up of endothelial derangement and therapeutic efficacy in Kawasaki disease (KD), circulating ACE, vWF routine hematological tests and cardiac involvement were assessed in 32 children with established diagnosis of KD before and up to six months after intravenous gamma globulins (i.v.IG) treatment. I.v.IG treatment normalized progressively all the hematological parameters to levels comparable with healthy controls within 30 days. At baseline, ACE levels resulted significantly lower (1.8 +/- 1.3 pmol/ml/min), and vWF levels significantly increased (210.3 +/- 35.2%) when compared with controls (respectively 7.0 +/- 0.9 pm/ml/min and 99 +/- 17.9%). Seven days after the treatment vWF levels were decreased (188 +/- 18.4%) but still significantly higher than controls, and fully normalized after 15 days (104.8 +/- 14.3%). ACE levels were found progressively increased at 7, 15, and 30 days after the treatment (respectively 2.7 +/- 1.0, 3.7 +/- 0.4, 5.04 +/- 0.9 pm/ml/min) and reached the range of normality only after two months (7.74 +/- 2.46 pm/ml/min). The present study shows that ACE and vWF circulating levels are significantly modified during the acute phase of the disease. PMID- 10365773 TI - The putative mechanistic basis for the modulatory role of endothelin-1 in the altered vascular tone induced by Trypanosoma cruzi. AB - Chagas' disease, caused by Trypanosoma cruzi, is an important cause of heart disease in Latin America. T. cruzi-induced microvascular compromise, in turn, is thought to play a major role in chagasic heart disease. Previous in vitro studies have implicated endothelin-1 (ET-1) as a potentially important vasomodulator present in increased levels in the supernatant of T. cruzi infected cultured human umbilical vein endothelial cells (HUVEC). Thus, the goal of the present investigation was to further evaluate the potentially important contribution of ET-1 to T. cruzi-induced alterations in vascular tone in vitro. Bioassay studies once again documented that exposure of isolated rat aortic rings to infected HUVEC supernatants elicited contractile responses whose steady-state magnitude was significantly greater than contractile responses elicited by exposure of aortic rings to uninfected HUVEC supernatants. Furthermore, the increased aortic contractility was significantly attenuated by the presence of the ET(A) subtype selective antagonists BMS-182,874 or BQ-123. Additionally, incubation of HUVEC with either verapamil or phosphoramidon prior to infection was also associated with reduced aortic contractility, upon exposure to the supernatant. Phosphoramidon, but not verapamil, produced a significant decrease in the measured ET-1 levels in the HUVEC supernatant. Consistent with the bioassay results, preincubation of Fura-2-loaded cultured rat aortic vascular smooth muscle cells with verapamil resulted in a near complete ablation of ET-1-induced transmembrane Ca2+ flux. Taken together, these data are consistent with the hypothesis that ET-1-induced vasoconstriction may play an important modulatory role in the vascular compromise characteristic of T. cruzi infection. PMID- 10365774 TI - Reduction of endothelial cell related TGFbeta activity by thiols. AB - Transforming growth factor beta (TGFbeta) may play an important role in diseases characterized by pulmonary fibrosis. We have previously demonstrated that thiols inhibit the pro-oxidant effects of TGFbeta1 in bovine pulmonary artery endothelial cells (BPAEC). To help define the mechanism of this observation we have examined the effect of reduced (GSH) and oxidized (GSSG) glutathione, N acetyl cysteine (NAC) and cysteine (CYS) on the biological activity of a) TGFbeta released by bovine pulmonary artery endothelial cells (BPAEC) into culture medium, and b) commercially available porcine platelet TGFbeta1. The biological activity of TGFbeta (following activation) released into the medium from cultured BPAEC was significantly reduced when the cells were cultured in the presence of 10 mM GSH or 10 mM NAC for 24 h (10 mM GSH: 85.7 +/- 50 pg/ml/10(6) cells and 10 mM NAC: 127.3 +/- 35 pg/ml/10(6) cells, compared with control: 541 +/- 8.9 pg/ml/10(6) cells; p < 0.05). Thiols (10 mM GSH, 10 mM NAC and 5 mM cysteine), added directly to cell-free conditioned medium or to a commercially available preparation of porcine platelet TGFbeta1 for 6-24 h had a similar inhibitory effect on the biological activity of TGFbeta and altered the structure of porcine platelet TGFbeta1 as determined by mass spectroscopy. These thiols failed to reduce the expression of TGFbeta mRNA in BPAEC as measured by a competitive polymerase chain reaction assay. Incubating endothelial cells or cell-free conditioned medium with GSSG did not alter the biological activity of TGFbeta. Lower doses of thiols (0.1-1 mM), that we have shown inhibit the antiproliferative and pro-oxidant effects of exogenous TGFbeta1 on BPAEC, had no direct effect on TGFbeta bioactivity. In summary, thiols are capable of reducing the effects of TGFbeta in biological systems through a direct effect on the TGFbeta molecule. However, this action appears to be dose-dependent, and at low doses (0.1-1 mM) thiols may also inhibit the actions of exogenous TGFbeta1 in cell culture through a mechanism involving the cellular redox status. PMID- 10365775 TI - A novel, 85 kDa endothelial antigen differentiates plasma membrane macrodomains in lung alveolar capillaries. AB - A monoclonal antibody raised against purified rat lung endothelial plasma membranes was found to recognize an apparently novel, 85 kD, integral endothelial plasma membrane glycoprotein. By immunofluorescence and electron microscope immunocytochemistry, this endothelial antigen was detected at the luminal and tissue fronts of all rat endothelia, except those of discontinuous type (liver and spleen sinusoids). In lung alveolar capillaries the antigen appeared to be uniquely associated with the very attenuated endothelial processes forming the blood-air barrier, and virtually absent on the surface of the rest of the cell, where the nucleus and the organelles are located. No other cells, except fibroblasts, appeared to be labeled by this monoclonal antibody. PMID- 10365776 TI - Macro and microheterogeneity in normal endothelial cells: differential composition of luminal glycocalyx and functional implications. AB - Endothelial cells (EC) are involved in various physiological and pathological processes through the expression of their surface glycoproteins. They are covered by the glycocalyx, composed of glucidic residues from cell surface membrane glycoproteins, glycoplipids and proteoglycans. Glucidic sequences can be specifically characterized by their binding to lectins. Eight lectins were used to investigate the distribution and regulation of EC surface glucidic residues in various blood vessels of adult and newborn pigs. EC lectin binding was compared to von Willebrand factor (vWF) expression as EC reference marker. Six out of eight lectins (BSI-B4, DBA, EEA, HP, MAL I and PNA) were helpful for this determination. Considering only the intensity of labelings, vWF and DBA gave the best stainings of adult pig ECs. In newborn pigs, the best labelings were obtained with EEA and MAL I. Furthermore, the distribution of lectin binding to ECs and EC vWF expression was heterogeneous depending on the EC location along vascular tree and age. Beside this macroheterogeneity this study highlights a microheterogeneity of EC lectin binding and vWF expression in situ, defined as a staining of equal intensity by individual ECs, scattered among negative ones, in a given vascular segment. EC surface sugar residues were differently modulated in newborn and adult pig ECs and differently according to EC vWF expression. The functional involvement of EC glycocalyx was reflected by EC lectin binding in the spleen and liver. This study emphasizes the high level of EC heterogeneity for various markers. The EC macro- and microheterogeneity reflect the "plasticity" or "unstability" of EC phenotypes and suggests that ECs are subject to several levels of regulation and are probably grouped in functional clusters to best adjust their functions to microenvironmental requirements. This concept must be considered in further investigations notably in in vitro studies where EC phenotype can be altered. PMID- 10365777 TI - Technical and clinical comparison of two fully automated methods for the immunoassay of CA 125 in serum. AB - The sensitivity and precision of two fully automated enzyme immunoassays, a chemiluminescent enzyme immunoassay (CLEIA) and an enzyme-linked immunosorbent assay (ELISA), for the determination of the ovarian carcinoma antigen CA 125 were evaluated by comparison with an immunoradiometric assay (IRMA). Sera were obtained from patients with ovarian carcinoma (N = 28 before treatment and N = 24 after treatment), digestive system cancer (N = 21 before treatment) and from healthy women (N = 90). The CLEIA showed a good agreement with the IRMA in terms of the positivity rate, accuracy and assay linearity, whereas the ELISA gave some false positive results. The mean value of CA 125 in the sera of healthy women was 14, 16 and 20 U/ml determined using the CLEIA, IRMA and ELISA procedures with standard deviations (SD) of 6.9, 7.3 and 8.8 U/ml, respectively. Both the reproducibility and precision of the CLEIA with coefficients of variation (CV) of 4.6% intra-assay and 7.6% inter-assay were better than those of the ELISA with CV of 6.2% intra-assay and 15.2% inter-assay (N = 16). We conclude that the CLEIA is the preferable method for CA 125 determinations and the diagnosis of ovarian carcinoma. PMID- 10365778 TI - Real-time analysis of integrin-mediated chemotactic migration of T lymphocytes within 3-D extracellular matrix-like gels. AB - We have developed a novel 3-D gel reconstituted with major extracellular matrix (ECM) glycoproteins to follow the dynamics of migration of human T cells locomoting, in real-time, on gradients formed by representative chemoattractants: the C-C chemokine RANTES, and the pro-inflammatory cytokine IL-2. In the absence of chemoattractants, none of the T cells migrated directionally and the levels of random migration or cell polarization were low. However, major fractions of T cells placed in IL-2 and RANTES gradients in the gels polarized immediately after exposure to the chemoattractants. Shortly after polarization, 25% of the T cells migrated, in either a random or directional fashion, towards the sources of the chemoattractants; additional 5-10% of the cells remained polarized but stationary. The number of T cells migrating directionally towards RANTES or IL-2 peaked along with the formation of the chemotactic gradients. The directional migration of T cells was increased by a short pre-exposure to low doses of IL-2, which did not alter the level of expression of the beta1 integrins. The directional migration of T cells towards IL-2 and RANTES was mediated by IL-2R and pertussis toxin-sensitive receptors, respectively, and the directional, and to a lesser degree, the random locomotion of T cells induced by both chemoattractants required intact tyrosine kinase signaling and activities of the alpha4, alpha5, and, to a lesser degree, the alpha2 and alpha6 members the beta1 integrins. Our system enables the real-time tracking of individual locomoting lymphocytes and the analysis of their dynamic interactions with ECM components and cytokines. PMID- 10365779 TI - Optimal storage conditions for preserving granulocyte viability as monitored by Annexin V binding in whole blood. AB - It is important in the laboratory to develop techniques to preserve leucocyte viability in blood specimens for subsequent flow cytometric analysis. This article describes a new simple whole blood lysis method using Annexin V FITC staining which can be used to define both early and late apoptosis of granulocytes in heterogeneous cell populations, without the need for additional stains or to purify the cells (which may result in loss of the cells of interest). The differential Annexin V binding assay was in good agreement with the light microscopy reference method and showed excellent correlation with 7 aminoactinomycin D (7-AAD) staining. It was not affected by problems of morphological interpretation and artifactal changes of granulocyte deformability noted using light microscopy, or the technical difficulties encountered due to red cell contamination using the 7-AAD method. Using this new differential Annexin V staining method, we determined the optimum conditions that maintain granulocyte viability for subsequent flow cytometric analysis and are now employed in our laboratory. These conditions were lithium heparin (Hep) anticoagulated whole blood specimens kept at 4 degrees C with the addition of nutrient medium. Specimens that are anticoagulated with acid citrate dextrose (ACD) or ethyl-diacetyl-tetraacetic acid (EDTA) should also be treated similarly to preserve granulocyte viability and to overcome problems associated with identification of cell populations by flow cytometry. PMID- 10365780 TI - Clonal expansion of antigen-specific CD8+ cytotoxic T lymphocytes is regulated by late exposure to serum to prevent apoptosis. AB - Although serum-free media have been used to expand lymphokine-activated killer cells, antigen-specific CD8 T cell cytotoxicity does not develop in vitro in the absence of serum. The immunodominant Vbeta17 response to an influenza A matrix protein epitope restricted by HLA A2.1 was used to study the serum requirement for CTL activation. Serum acts directly on T cells and not indirectly by activating APCs. In the absence of serum, the initial steps of T cell activation, including expression of CD69 and CD25, are unimpaired and some antigen-specific cytotoxicity may be generated in the first few days after stimulation. However, expression of late activation markers, such as HLA-DR and CD38, and clonal expansion of class I-restricted antigen-specific CTL does not occur if CTL are not exposed to serum within 4 days of antigen exposure. The antigen-specific CTL, but not unstimulated bystander T cells, undergo apoptosis if they are not exposed to serum within a few days of activation. Apoptosis of TCR-activated CTL does not appear to be Fas-mediated since it is not blocked by inhibiting the Fas pathway. Therefore, late exposure to an unidentified serum protein regulates the clonal expansion of TCR-activated CD8 CTL. PMID- 10365781 TI - Quantitative flow cytometry for the analysis of T cell receptor Vbeta chain expression. AB - Detailed characterisation of the T cell receptor (TCR) repertoire expressed by peripheral blood lymphocytes has been used to study specific T cell responses in disease conditions. The methods have mostly involved molecular biology analysis of transcribed gene products isolated from T cell subsets or individual clones. Extensive characterisation of the TCR Vbeta chain repertoire by flow cytometry is now possible due to the recently increased availability of specific monoclonal antibodies. However, there are major logistical problems inherent in this analysis relating to the number of cells required to obtain accurate results and the vast amounts of data generated. To reduce these factors to a practical level, we have performed a detailed study to define the limits of precision of cell subset analysis by flow cytometry. Maximal achievable precision was obtained by analysing 10(4) lymphocytes; no significant improvement was obtained by analysing greater numbers of cells up to 10(5) cells, even for cell subsets present at frequencies as low as 0.5%. Careful application of these precision profiles will also permit more effective use of clinical research samples for flow cytometry when the availability of cells is limited. PMID- 10365782 TI - Elimination of non-viable 6-thioguanine-sensitive T cells from viable T cells prior to PCR analysis. AB - The study of T cell clones at the genomic level is expanding our understanding of their role in diseases such as rheumatoid arthritis (RA) and multiple sclerosis (MS). We have been carrying out genotypic analysis by PCR of hypoxanthine phosphoribosyltransferase (hprt) mutations in these cells. Mutant T cells in the population can be cloned on the basis of their resistance to the cytotoxic drug, 6-thioguanine-(6-TG). A difficulty is that the majority of primary human T cells are capable of only limited growth ex vivo, even in the presence of 'feeder' cells. PCR analysis of DNA from such clones is made difficult by the limited number of viable mutant (drug-resistant) T cells and the large number of dead (drug-sensitive) mononuclear cells and feeder cells. DNA from the 'dead' cells remains sufficiently intact for many weeks in culture and can represent a significant source of background in PCR analysis. Here we describe a method employing hypotonic shock and micrococcal nuclease that reliably eliminates non viable 6-TG-sensitive cells, allowing the study of the hprt gene in < 200 T cells by PCR. PMID- 10365783 TI - An immunoassay for assessment of receptor tyrosine kinase autophosphorylation. AB - We have developed an immunoassay that can be used to assess autophosphorylation activity of receptor tyrosine kinases, yet does not require the use of synthetic peptide substrates or anti-receptor antibodies. In the assay described here, receptor autophosphorylation and detection take place entirely within the wells of 96 well microplates coated with Protein G and an anti-phosphotyrosine antibody. As the kinase reaction takes place, the antibodies capture the phosphorylated products in situ. Phosphorylation levels of captured receptors are measured by scintillation counting. Assay parameters were validated using A431 cell extracts containing EGF Receptor. The autophosphorylation capture assay is a simple and rapid method which can be adapted for use with robotics for qualitative HTS of potential inhibitors to any tyrosine kinase of interest. PMID- 10365784 TI - Growth of Mycobacterium bovis, Bacille Calmette-Guerin, within human monocytes macrophages cultured in serum-free medium. AB - Mycobacterium bovis BCG is an opportunistic agent that may be responsible for disseminated disease in immunocompromised individuals. Under physiological conditions, macrophages are the natural hosts and final killers of BCG. In the context of inherited or acquired immune disorders underlying disseminated BCG infections, macrophages fail to eradicate BCG or even to restrict its intracellular growth. The direct contribution of macrophages, in this setting of impaired BCG destruction, probably depends on the type of underlying immune deficiency and remains to be experimentally investigated. As an initial approach, we document here the fate of BCG within human monocytes and human monocyte derived macrophages (MDMs) cultured in commercially available serum-free medium (M-SFM). This medium was used to avoid potential problems associated with human or animal serum-supplemented medium. We show here that both monocytes and MDMs cultured in M-SFM display the morphological features and functional activities expected for such cells. We also show that after an initial phase of intracellular destruction, BCG grow within infected monocytes-macrophages, as shown by colony forming unit (CFU) counts and Ziehl-Nielsen staining. By an electron microscopic analysis, we show that the BCG always reside within phagosomes and that 24-h postinfection many phagosomes stain for the hydrolytic enzyme acid phosphatase. Finally, we compare bacterial growth in vitro within phagocytes from healthy individuals and patients with chronic granulomatous disease (CGD), an inheritable condition associated with disseminated BCG infection in vivo. No destruction of intracellular BCG was achieved by the patients cells, revealing the essential mycobactericidal role of the respiratory burst in human phagocytes. Investigations of BCG growth within MDM cultured in M SFM from patients with other conditions which predispose to clinical BCG infection is therefore warranted. PMID- 10365785 TI - Isoform specificity of commercially-available anti-TGF-beta antibodies. AB - The transforming growth factor-beta (TGF-beta) cytokine family has important and complex effects on many biologic processes. Mammals have three TGF-beta isoforms which differ in their primary amino acid sequence, receptor binding characteristics, distribution, and function. Characterization of TGF-beta production and localization is critically dependent upon appropriate reagents, including antibodies. We have analyzed the isoform specificity of eight commercially-available TGF-beta antibodies, including one monoclonal antibody and seven polyclonal antibodies. We carried out semi-quantitative Western blot analysis using recombinant TGF-beta1, beta2, and beta3 as targets. We found that sensitivity and isoform specificity are dependent in part upon the presence or absence of reducing conditions. The antibodies tested showed a broad range of sensitivity, with an ability to detect 50 pg to 20 ng. Cross-reactivity with another, incorrect isoform was seen with several antibodies, and ranged from 0.2% to 42%. Nevertheless, we identified TGF-beta antibodies directed against each isoform which provide moderate-to-high sensitivity and specificity when used in Western blot analysis. These results may have relevance for investigators who wish to detect particular TGF-beta isoforms with techniques other than Western blot analysis, particularly when these techniques involve denatured proteins. PMID- 10365787 TI - Influenza virus: a novel method to assess viral and neutralizing antibody titers in vitro. AB - The present report describes novel in vitro assays to determine influenza virus titers and virus neutralizing antibody levels. For determination of viral titers, serial dilutions of influenza virus were incubated with MDCK-cells and cultured for 48 h under a methylcellulose overlay in 24 well plates. Cells were fixed, permeabilized and stained with a monoclonal antibody specific for hemagglutitin (HA) and a peroxidase labelled second stage antibody. The sensitivity of the assay was 100-1000 times greater than a conventional hemagglutination test using fresh chicken blood. For determination of influenza virus neutralizing activity, viral samples were incubated with serial dilutions of antibody and residual viral activity was assessed in 96 well plates by the same procedure as described above. This assay made it possible to distinguish between IgM and IgG antibody titers and was about 5-10 fold more sensitive than a classical hemagglutination inhibition assay using fresh chicken blood. PMID- 10365786 TI - High resolution affinity chromatography of an anti-steroid antiserum by gradient elution with propionic acid. AB - Chromatographic resolution of polyclonal antibodies is a challenging analytical problem and a successful method may have many applications. We have resolved polyclonal antibodies against testosterone 3-(O-carboxymethyl)oxime on a homologous testosterone-Sepharose matrix by using a ternary gradient system of propionate-1 M propionic acid-2 M propionic acid. Nineteen peaks were detected, sixteen of which were characterized by steroid binding activity and IgG assay. The relative affinities of the pooled fractions, determined by ELISA and by thiocyanate elution, correlated well with their retention times. The slopes of the displacement curves in antigen-immobilized testosterone ELISA increased with retention times of the fractions; testosterone required for 50% displacement being, 90, 5.8 and 24 ng/well for fraction I, fraction XVI and total antiserum, respectively. Cross-reactivity of the fractions towards 5alpha dihydrotestosterone varied from 31 to 74% compared to 47% in the total antiserum. PMID- 10365788 TI - Design of limiting dilution analysis experiments for helper T lymphocyte precursor frequency determination in the context of allogeneic bone marrow transplantation. AB - Helper, interleukin 2 (IL-2) producing, T lymphocyte precursor (HTLp) frequency determination by limiting dilution analysis (LDA) is of value for quantifying alloreactivity in allogeneic bone marrow transplantation (BMT). LDA assays are labour-intensive and time-consuming to perform and the numbers of donor and recipient cells available are limited. It is therefore important that the design of the experiment yields reliable frequencies with a minimum of effort and a realistic cell requirement. We have critically evaluated the methods proposed for LDA design by Strijbosch et al. [Strijbosch, L.W., Buurman, W.A., Does, R.J., Zinken, P.H., Groenewegen, G., 1987. Limiting dilution assays. Experimental design and statistical analysis. J. Immunol. Methods 97, 133] and by Blackett and Gordon [Blackett, N.M., Gordon, M.Y., 1996. Optimizing limiting dilution assays: frequency and 'ability' measurements of haemopoietic progenitor cells. Br. J. Haematol. 92, 507 (see comments)] and found them inadequate for this application. The estimation of the HTLp frequency is traditionally based on the single-hit Poisson model and the adequacy of this model was compared with that of a double hit model. The results were in favour of the single-hit model. Ten different LDA experimental designs were explored by Monte Carlo simulations. The optimal design exploits the maximal numbers of cells that can be obtained for analysis to estimate HTLp frequencies in the range 1:1,000,000-1:20,000 with a coefficient of variation of 10-20% and with a minimum of manual labour. PMID- 10365789 TI - Quantitation of human haptoglobin: comparative ELISA studies using adsorption and capture methods. AB - Three ELISA methods for the quantitation of haptoglobin (Hp) in plasma and albumin are described: a polystyrene direct adsorption method and capture methods with antibody and hemoglobin. Hp aggregates generated by 60 degrees C heating showed as much as a hundred-fold higher response by polystyrene adsorption compared to the two capture methods, while unheated Hp showed comparable responses by the three methods. PMID- 10365790 TI - Cytokine and immunoglobulin concentrations in cervical secretions: reproducibility of the Weck-cel collection instrument and correlates of immune measures. AB - Elucidation of local immune response at the cervix is important for understanding and evaluating STD vaccine approaches currently being proposed. However, no well validated method exists for the collection of cervical secretions for evaluation of cervical immune response. The purpose of this study was to determine the reproducibility of the Weck-cel sponge used to collect cervical secretions for immunological assessment. Additionally, it was possible to examine correlates of immunity as part of our investigation. Two cervical secretion specimens were collected sequentially from each of 120 women using Weck-cel sponges. Cervical secretions were collected prior to Pap smear sampling to avoid blood contamination. At the laboratory, the duplicate specimens were weighed and tested in replicate wells to determine the concentration of two cytokines (IL-10 and IL 12) and two immunoglobulin isotypes (IgG and IgA). IL-12, total IgG, and total IgA showed a strong correlation between samples from the same woman ranging from 0.78 to 0.84. Kappa coefficients obtained after categorizing assay results ranged from 0.62 to 0.67. Variance components analysis suggested that 69% to 85% of the variance observed was accounted for by between-women variance, with the remaining variability attributed to variation between samples collected from the same woman. IL-10 results were less reproducible than those obtained from the other assays examined, suggesting problems with the assay used to measure this cytokine rather than with the Weck-cel sampling instrument. Various factors were found to significantly correlate with cytokine and immunoglobulin measures at the cervix. Age and reproductive status were associated with all four immune measures; women over 50 years of age and those who were postmenopausal had increased concentrations of IL-10, IL-12, IgG, and IgA. Hemoglobin concentrations were positively correlated with IgG and IL-10 concentrations, but not with IgA or IL 12 concentrations, suggesting local production of IgA and IL-12. The concentration of all immune measures decreased with increasing volume of collection. No significant association was observed between time from collection to freezing of specimens and concentrations of cytokines or immunoglobulins. Overall, our data suggest that measurement of immunological parameters in cervical secretions collected using Weck-cel sponges are reproducible. In addition, various correlates of cytokine and immunoglobulin concentrations were identified. PMID- 10365791 TI - A sandwich enzyme-linked immunoabsorbent assay for measurement of picogram quantities of murine granulocyte colony-stimulating factor. AB - Granulocyte colony-stimulating factor (G-CSF) stimulates the proliferation and differentiation of hematopoietic progenitor cells of the neutrophil lineage. Measurement of murine G-CSF levels will allow examination of its role in host defense using murine models. Therefore, we developed a sensitive sandwich enzyme linked immunoabsorbent assay (ELISA) for murine G-CSF. A polyclonal antibody to recombinant murine G-CSF was produced in rabbits and isolated using a protein A column. This purified native IgG served as the capture antibody and a portion of the IgG was biotinylated to serve as the developing antibody. Specificity was verified by lack of reactivity to GM-CSF, IL-6, IL-3, prolactin, and growth hormone. The lower limit of sensitivity routinely extended to 16 pg/ml in multiple ELISAs. Intra-assay coefficient of variation (CV) ranged from 3.4 to 21.5% across the detection limits of the assay, with the greatest variance occurring near the standard curve maximum. Interassay CV ranged from 11.5 to 23.3%. The ability of the ELISA to detect G-CSF in different sample preparations was examined in RPMI 1640 with 10% FCS, Hanks balanced salt solution, PBS/Tween 20/2% FCS, and the dilution media for ELISA (10% BLOTTO/PBS/0.05% Tween-20). Average recovery in these media ranged from 98 to 107%. Heparin anti-coagulated normal mouse plasma had a suppressive effect on the ELISA that varied between individual mice. Recovery was also determined from liver, spleen, and lung homogenate suspensions at dilutions of 1:5, 1:10, and 1:20 in dilution buffer. Recovery from liver was optimal at the 1:10 and 1:20 dilutions at 105%, with that of the 1:5 dilution at 135%. Recovery from spleen ranged from 94 to 96%. Lung homogenate displayed enhanced recovery (139% or greater) across all dilutions. The ability of the assay to detect G-CSF was explored by measurement of G-CSF levels in peritoneal lavage following polymicrobial intra-abdominal infection. Peak levels of G-CSF production occurred at 16 h after cecal ligation and puncture surgery with 18- and 21-guage needles (75.7 ng/ml and 111.4 ng/ml, respectively) as compared to the sham animals (0.61 ng/ml). The assay was found to be specific, sensitive, and accurate for measurement of murine G-CSF in a variety of sample types. PMID- 10365792 TI - Assessment of an automated solid phase competitive fluoroimmunoassay for benzoylecgonine in untreated urine. AB - A new solid phase fluoroimmunoassay using a fully automated flow fluorometer adapted for urinalysis of drug metabolites is described. Fluorescein-conjugated benzoylecgonine (FL-BE) and monoclonal antibodies (mAb) against benzoylecgonine (BE) were the reagents used for demonstration. The solid phase consisted of anti BE mAbs immobilized on the surface of polymethyl methacrylate (PMMA) beads. Free BE in solution competed with FL-BE and reduced bead-bound fluorescence in a concentration-dependent manner. The binding of FL-BE to the anti-BE mAb reached steady-state within minutes. FL-BE was not bound by uncoated beads nor beads coated with non-specific proteins or IgG. The signal-to-noise ratio was 33, and the sensitivity of the assay was 2 ng ml(-1) for BE. The effective concentration of BE was 1 to 100 ng ml(-1), with an IC50 value of 12 ng ml(-1). The mAb showed equal affinities for BE, cocaine, and cocaethylene, but a five order-of-magnitude lower affinity for ecgonine and ecgonine methylester. In a double-blind comparison using clinical urine samples, the data from this single-step competitive assay had excellent agreement with results obtained using a fiber optic biosensor (FOB), and the EMIT assay performed commercially. The assay provided kinetic data rapidly and can be used to detect small analytes for which antibodies and fluorescein conjugates are available. The affinity of the mAb for FL-BE, calculated from kinetic analysis of the time course of the on and off reaction, was 2.25 x 10(-9) M. PMID- 10365793 TI - Biodistribution of filamentous phage-Fab in nude mice. AB - In vivo panning of peptide libraries in mice has allowed the isolation of peptides which target the vasculature of specific organs. The application of this approach to phage displaying Fab fragments (phage-Fab) could lead to the isolation of antibodies which recognize novel tumor antigens. In this study, we have evaluated the biodistribution of phage-Fab in nude mice. Balb/c nude mice were injected intravenously with 10(9) TU of phage displaying the anti-colon cancer Fab c30.6. Blood samples were collected at nine time points over a period of 72 h and three groups of four mice were sacrificed at 4 min, 24 h and 72 h. Normal tissues (liver, colon, spleen, kidneys, lungs, skeletal muscle) and faeces were collected at these time points and the number of viable phage in each sample was determined. The distribution of phage in tissues was also examined by immunohistochemical analysis of paraffin-embedded tissues. Regression analysis of plasma kinetic data showed that the half-life and the volume of distribution of phage was 3.6 h and 1 ml, respectively. Phage uptake occurred predominantly in lungs, kidneys, spleen and liver. Relatively few phage were distributed to colon and muscle, and phage were eliminated from the circulation by 72 h. Immunohistochemical analysis showed phage to be mainly within the vasculature at 4 min, whereas notable phage extravasation was observed at 24 h and 72 h. In conclusion, this study provides information on the in vivo behavior of phage-Fab which will be useful in the design of in vivo panning strategies. By choosing appropriate time points for tissue collection, it may be possible to isolate novel Fabs against both intra- and extravascular targets. PMID- 10365794 TI - Use of fixed cells in cell contact-dependent cytotoxicity assays for TNF: a cautionary report. AB - The use of fixed effector cells or cell membrane preparations in assays to study cell surface TNF-mediated immunological effects has been widespread for more than a decade. The assumption has always been made that observed effects of fixed cells are due to the cell surface TNF molecule. Here we report that paraformaldehyde-fixed, LPS-stimulated RAW264.7 cells release a factor that causes cytotoxicity against TNF sensitive WEHI-164 cells. The factor was neutralized by soluble TNF receptors as well as antibody and could not be removed by ultracentrifugation at 100,000 x g, and is therefore likely to be a form of soluble TNF. This has important implications for the interpretation of these assays, given that cell surface TNF is thought to exert biological effects through a different signaling mechanism than soluble TNF. PMID- 10365795 TI - Improving the expression of chimeric antibodies following homologous recombination in hybridoma cells. AB - Mouse/human chimeric antibodies can easily be generated by exchanging the immunoglobulin constant gene segments with human sequences in mouse hybridoma cells by gene targeting. This obviates the need to isolate the variable gene regions from the hybridoma and permits high-level expression of chimeric antibodies, because the production rate of the hybridoma is often maintained. Here we show that the efficiency of this strategy can be further improved by increasing the number of high-producer clones arising from a recombination experiment. In principle, antibody production can be enhanced by removing the selection marker genes from the targeted cells. PMID- 10365796 TI - Histological changes in liver biopsies after one year of lamivudine treatment in patients with chronic hepatitis B infection. AB - BACKGROUND/AIMS: The aim of this study was to examine the histological changes in liver biopsies induced by 52 weeks of lamivudine therapy in patients with e antigen positive and e-antigen negative chronic hepatitis B infection. METHODS: Twenty patients were enrolled into this open-label study. All patients had a liver biopsy within the 4 weeks before starting lamivudine therapy. Lamivudine was given orally at a dose of 100 mg OD for 52 weeks. A second liver biopsy was taken for comparison at the end of week 52. Blinded biopsies were evaluated by a histopathologist and scored according to Knodell's histology activity index (HAI). RESULTS: Ninety-five percent (19/20) patients had a reduction of their hepatic necroinflammatory HAI score (components 1 through 3) by > or =2 points at the end of 52 weeks of lamivudine therapy compared to their pretreatment values. Not only were improvements in necroinflammatory activity observed, but 7/20 (35%) of patients had improvement in fibrosis. This histologic improvement was independent of the presence or absence of e-antigen. CONCLUSIONS: Significant improvements in liver histology can be obtained in the majority of patients when they are treated with lamivudine for 1 year. PMID- 10365797 TI - Absence of mutations in the YMDD motif/B region of the hepatitis B virus polymerase in famciclovir therapy failure. AB - BACKGROUND/AIMS: Nucleoside analogues such as lamivudine and famciclovir are potent drugs for treatment of chronic hepatitis B virus infection. Breakthrough infections during lamivudine therapy are associated with mutations in the YMDD motif and putative B region of the HBV polymerase. This study investigated whether failure of famciclovir therapy is also associated with presence or emergence of particular mutations in the HBV polymerase. METHODS: We analyzed longitudinally the sequence of the priming and polymerase domain in seven patients with primary non-response to therapy and two patients with a breakthrough during therapy. Two patients who responded to therapy served as a control. RESULTS: The YMDD motif and the B region were conserved in all isolates. V-->I changes at position 555 just downstream of the YMDD motif were observed before and during therapy in a virus subpopulation of two patients with a primary non-response. In patients with a breakthrough, 378-V-->I and 424-N-->D mutations emerged in the N terminal part of the polymerase domain during follow-up. Lamivudine rescue therapy initiated in four patients, including a patient infected with YMDD(555-V-->I) variants, efficiently reduced viremia. CONCLUSIONS: These data indicate that failure of famciclovir therapy can occur independently of mutations in the YMDD motif or B region of the HBV polymerase and provide a rationale for rescue therapy with lamivudine. PMID- 10365799 TI - Risk of hepatitis C virus transmission to surgeons and nurses from infected patients: model-based estimates in France. AB - BACKGROUND/AIM: The aim of this study was to estimate the annual number of cases of hepatitis C virus transmission from infected patients to uninfected surgeons or nurses due to percutaneous injury during invasive procedures. METHODS: The risk of transmission was estimated using a model involving three probabilities: A, that a health care worker sustains at least one percutaneous injury during a procedure; B, that 1 to 10% of patients are seropositive for hepatitis C virus; and C, that infection by this virus is transmitted to the Health Care Worker after such exposure. Probability A was estimated from the results of 2 French multicentric prospective trials. Probability C was estimated from the results of 9 international prospective studies. A ten-fold decreased risk was assumed for surgeons who wear gloves and use solid-bore suture needles. RESULTS: During a single procedure, the estimated probability of hepatitis C virus transmission from an infected patient to an uninfected surgeon ranged from 4.2x10(-5)% to 4.2x10(-4)%, and from 2.98x10(-6)% to 2.98x10(-5)% to an uninfected nurse. For surgeons, the estimated annual cumulative risk of occupational infection ranged from 0.01% to 0.1% (1 in 10000 to 1 in 1000), and for nurses from 0.0054% to 0.054% (1 in 18700 to 1 in 1900). CONCLUSIONS: Between 2 and 21 surgeons out of a total 20000 are estimated to acquire occupationally-related hepatitis C virus infection, and between 16 and 167 nurses out of a total 300000. These estimates strongly justify introducing preventive measures to protect health care workers from bloodborne infection. PMID- 10365798 TI - Mechanism of action of vaccine therapy in murine hepatitis B virus carriers: vaccine-induced activation of antigen presenting dendritic cells. AB - BACKGROUND/AIMS: Vaccine therapy in which vaccine containing hepatitis B surface antigen (HBsAg) is injected has shown therapeutic activity (vaccine therapy) in some human and murine chronic hepatitis B virus (HBV)-carriers. Using HBV transgenic mice (HBV-Tg), an animal model of the HBV-carrier state, the mechanism underlying the antiviral and immune modulatory capacity of vaccine therapy was studied. METHODS: Placebo-controlled, double-blind trials of vaccine therapy were conducted in HBV-Tg; some HBV-Tg responded to the therapy, whereas others were non-responders. The titers of HBV-markers, the functions of lymphocytes and antigen presenting dendritic cells were compared between vaccine responders and vaccine non-responders. RESULTS: The prevaccinated titers of HBsAg, hepatitis B e antigen (HBeAg), HBV DNA and the responses of lymphocytes to polyclonal mitogens were almost unchanged between responders and non-responders, but the levels of proliferation of HBsAg-specific lymphocytes from non-responders was significantly lower than responders (p<0.05). The capacity of dendritic cells to induce proliferation of T cells and production of antibody to HBsAg (anti-HBs) was significantly higher in responders compared with non-responders (p<0.05). Injection with HBsAg resulted in upregulation of MHC class II and CD86 antigens (p<0.05) on dendritic cells and increased production of IL-12, IL-2 and TNF-alpha in cultures (p<0.05) in vaccine responders but not in non-responders. CONCLUSIONS: The activation of dendritic cells following injection with vaccine containing HBsAg is the vital factor underlying the therapeutic potentiality of vaccine therapy in HBV carriers. PMID- 10365800 TI - Prevalence of hepatitis C virus infection in patients with antiphospholipid syndrome. AB - BACKGROUND/AIMS: The aim of this study was to determine the prevalence and clinical significance of hepatitis C virus (HCV) infection in patients with the antiphospholipid syndrome (APS). METHODS: A series of 88 consecutive patients (78 female and 10 male), with a mean age of 39 years (range 15-79), was prospectively studied. All patients had been diagnosed with APS: 54 (61%) primary APS and 34 (39%) APS associated with systemic lupus erythematosus. A group of 200 apparently healthly blood donors was included in the study. Anti-HCV antibodies were investigated in the serum of all patients using a third-generation ELISA and confirmed by recombinant immunoblot assay. RNA-HCV was investigated in anti-HCV positive samples by polymerase chain reaction. Anticardiolipin, anti-beta2 glycoprotein I and antiprothrombin antibodies were evaluated by ELISA. Lupus anticoagulant was studied by coagulometric assays. RESULTS: Only 2 (2.2%) patients showed positivity for anti-HCV antibodies, but none of them had clinical or biochemical signs of liver disease. Furthermore, RNA-HCV was not detected in serum of any of these patients. Lupus anticoagulant was positive in 57% of patients. Anticardiolipin antibodies were positive in 60% of patients, anti-beta2 glycoprotein I antibodies in 43% of patients, and antiprothrombin antibodies in 56% of patients. The prevalence of anti-HCV in blood donors was 1%. CONCLUSIONS: The prevalence of anti-HCV in patients with APS is low and similar to that in healthy people in our area. HCV infection does not seem to be involved in the etiopathogenesis of this syndrome. PMID- 10365801 TI - Serum ferritin and hepatic glutathione concentrations in chronic hepatitis C patients related to the hepatitis C virus genotype. AB - BACKGROUND/AIMS: Increased serum ferritin is thought to be responsible for activation of glutathione turnover in patients with chronic hepatitis C. The aim of the study was to evaluate a possible correlation between levels of serum ferritin and concentrations of hepatic, plasmatic and lymphocytic glutathione in a selected cohort of chronic hepatitis C patients in relation to the hepatitis C virus genotype. METHODS: The study considered 130 chronic hepatitis C patients and 23 control subjects. Hepatic glutathione was determined from biopsy liver specimens by high performance liquid chromatography. Total Iron Score was assessed by scoring iron separately within hepatocytes, sinusoidal cells and portal tracts. Blood samples were tested for determination of serum ferritin, and plasmatic and lymphocytic glutathione levels. Hepatic and erythocyte malonyldialdehyde were also determined along with peripheral blood mononuclear cell cytotoxic assay. RESULTS: Patients with genotype 1b showed higher levels of serum ferritin compared to patients with genotype 2a/2c and 3a and to controls, along with a significant reduction of the concentrations of hepatic, plasmatic and lymphocytic glutathione and peripheral blood mononuclear cell cytotoxic activity. The levels of serum ferritin correlated significantly to Total Iron Score, to hepatic, plasmatic and lymphocytic glutathione, to hepatic and erythrocyte malonyldialdehyde and to peripheral blood mononuclear cell cytotoxic activity. CONCLUSIONS: The levels of serum ferritin correlate significantly to lipoperoxidation markers in chronic hepatitis C patients. The increased production of free radicals with a reduced peripheral blood mononuclear cell cytotoxic activity may represent, especially in patients with genotype 1b, a factor underlying the resistance to interferon therapy and may influence the evolution of the liver disease by enhancement of the cytopathic effect of hepatitis C virus. PMID- 10365802 TI - Long-term follow-up of chronic hepatitis C patients with sustained virological response to alpha-interferon. AB - BACKGROUND/AIMS: This study aimed to determine the long-term outcome of hepatitis C virus (HCV)-infected patients who respond to interferon treatment with clearance of serum HCV RNA. METHODS: We performed a long-term biochemical, virological, and histological follow-up of all sustained virological responders, defined as those who became HCV RNA negative at follow-up 6 months after the end of treatment, from 3 controlled interferon trials performed in Sweden between 1988 and 1994. RESULTS: At biochemical and virological long-term follow-up performed in 26 sustained virological responders 3.5-8.8 years (mean +/- SD, 5.4+/-1.6 years) after the end of IFN therapy, 22 patients (85%) had normal serum ALT levels, and 24 patients (92%) were HCV RNA negative in serum. Liver biopsies performed in 23 patients 2.1-8.7 years (mean +/- SD, 5.0+/-1.8 years) after end of treatment showed no or minimal inflammation, whereas mild and probably irreversible fibrosis was seen in a few patients. CONCLUSION: In this well defined material of sustained responders to IFN therapy, the long-term prognosis was excellent. Nearly all had a durable response, not only biochemically and virologically, but more importantly also histologically with normalisation or near normalisation of previous histological lesions. PMID- 10365803 TI - Interferon-alpha plus ribavirin in chronic hepatitis C resistant to previous interferon-alpha course: results of a randomized multicenter trial. AB - BACKGROUND/AIMS: Interferon-alpha plus ribavirin seem to be more efficacious than interferon monotherapy in chronic hepatitis C. In a multicenter randomized trial, we evaluated the efficacy of this association for interferon-alpha resistant chronic hepatitis C. METHODS: Fifty patients who were non-responders to recombinant or lymphoblastoid interferon-alpha were randomized to receive either ribavirin (800 mg/day) plus leucocytic interferon-alpha (3 mega units thrice weekly) or the same dose of interferon-alpha alone, for 6 months. Effects of therapy were evaluated by serum aminotransferase and hepatitis C virus RNA levels and control liver biopsies. RESULTS: At the end of treatment, aminotransferase levels become normal in 9/26 patients receiving combination therapy (35% [confidence interval, 16% to 53%]) and in 2/24 receiving interferon-alpha alone (8% [confidence interval, -3% to 19%]) (p = 0.03). Aminotransferase normalization was never associated with hepatitis C virus RNA clearance. All patients with normal aminotransferase relapsed after discontinuation of therapy. At the end of treatment, mean hepatitis C virus RNA levels significantly decreased only in the group receiving combination therapy, but returned to pretreatment values 6 months thereafter. No histological improvement was observed in either group. CONCLUSIONS: There is no indication for treatment with interferon-alpha at the dose of 3 mega units thrice weekly plus 800 mg/day of ribavirin for 6 months in chronic hepatitis C resistant to interferon-alpha. PMID- 10365804 TI - HLA antigens in patients with interferon-alpha-induced autoimmune thyroid disorders in chronic hepatitis C. AB - BACKGROUND/AIMS: To determine the immunological predisposition to autoimmune thyroid disorders induced by interferon-alpha therapy, human leukocyte antigen (HLA) was analyzed in patients with chronic hepatitis C who developed autoimmune thyroid disorders during or after treatment with interferon-alpha. METHODS: Four hundred and thirty-nine patients with chronic hepatitis C (278 males and 161 females, aged 20-73 years) were treated with interferon-alpha (natural-alpha, 169; alpha-2a, 82; alpha-2b, 188) for 24 weeks. RESULTS: Seventeen of 439 (3.9%) patients developed symptomatic autoimmune thyroid disorders; these included nine cases of hyperthyroidism and eight cases of hypothyroidism. The incidence of HLA A2, B46 and Cw7 increased in patients with interferon-alpha-induced autoimmune thyroid disorders. Especially, the incidence of HLA-A2 (15/17; 88.2%) was significantly higher than that observed in the general population in Japan (corrected p-value (p(c)): p(c)<0.003). The odds ratios for the relative risk of the autoimmune thyroid disorders were A2, 10.6 [95% confidence interval, 2.4 46.5]; B46, 4.8 [1.6-14.0]; and Cw7, 3.0 [1.1-7.9]. CONCLUSIONS: Our study revealed that HLA-A2 is highly linked to the autoimmune thyroid disorders induced by interferon-alpha-therapy in patients with chronic hepatitis C. PMID- 10365805 TI - Possible association between serum GB virus C RNA level and disease activity in fulminant hepatitis type G. AB - BACKGROUND/AIMS: Whether GB virus C causes serious liver diseases remains controversial. The aim of the present study was to determine whether there is an etiological relationship between GB virus C and fulminant hepatitis. METHODS: The level of GB virus C RNA in the sera of three patients with fulminant hepatitis was quantitatively determined using the newly developed real-time detection polymerase chain reaction method, which is based on Taq Man chemistry. The NS 3 region of the viral genome isolated from the sera was sequenced at several time points to confirm whether the same virus was responsible for fulminant hepatitis during the patients' clinical courses. RESULTS: The sensitivity of the PCR was comparable to that of nested PCR and a linear relationship between RNA copy number and threshold cycle was observed for 10(1) and 10(6) RNA copies/ml (r = 0.99). The serum level of GB virus C RNA closely paralleled that of ALT in all patients. Sequence analysis of the NS3 region isolated from the patients' sera revealed that the same GB virus C strain infected the patients during their entire clinical courses, despite plasma exchange therapy. CONCLUSIONS: These observations suggest that GB virus C may be etiologically associated with fulminant hepatic failure, and is not merely an inactive bystander introduced by therapeutic plasma exchange. PMID- 10365806 TI - Activation of Kupffer cells and caspase-3 involved in rat hepatocyte apoptosis induced by endotoxin. AB - BACKGROUND/AIMS: Sepsis and lipopolysaccharides (LPS) cause mild to severe hepatic dysfunction. In this study, Kupffer cell activation, involvement of TNFalpha and caspases downstream of the TNFalpha receptor were examined in hepatocyte apoptosis induced by LPS. METHODS: In in vivo experiments, male Sprague-Dawley rats were injected intravenously with LPS, and small amounts of the blood and liver were sampled to evaluate apoptosis. Kupffer cells were inactivated by pretreatment with gadolinium chloride for 2 days. In in vitro experiments, hepatocytes and Kupffer cells were separately isolated from rat livers using collagenase perfusion. RESULTS: LPS induced time-dependent and dose dependent increases in the number of TUNEL-positive cells, which coincided with the apoptotic features of hepatocytes demonstrated by electron microscopy and DNA ladder. Activation of caspase-3-like proteases was observed with an increase in the number of apoptotic hepatocytes. Immunostaining with activated caspase-3 specific antibody showed that caspase-3 was activated only in the cytoplasm of TUNEL-positive hepatocytes. Inactivation of Kupffer cells by gadolinium chloride was concomitantly accompanied by the prevention of caspase-3 activation, hepatocyte apoptosis and liver injury induced by LPS. The co-culture system of hepatocytes and Kupffer cells, but neither cell culture system, individually, showed LPS-induced hepatocyte apoptosis. Kupffer cell-conditioned medium induced hepatocyte apoptosis, whereas addition of anti-TNFalpha antibody to Kupffer cell conditioned medium did not. Additions of acetyl-DEVD-CHO, acetyl-YVAD-CHO, and acetyl-IETD-CHO to Kupffer cell-conditioned medium decreased the number of apoptotic hepatocytes. CONCLUSIONS: These results suggest that the activation of Kupffer cells, TNFalpha and caspases downstream of TNFR1 were involved in hepatocyte apoptosis induced by LPS. PMID- 10365807 TI - Hepatic amino- to urea-N clearance and forearm amino-N exchange during hypoglycemic and euglycemic hyperinsulinemia in normal man. AB - BACKGROUND/AIMS: Hypoglycemia has well-described effects on glucose metabolism, whereas the possible effects on hepatic amino nitrogen conversion in relation to muscle amino nitrogen flux are more uncertain. METHODS: We studied six healthy young male subjects three times, i.e. for 6 h in the basal state, during a 6-h euglycemic hyperinsulinemic (1.5 mU/kg/min) clamp and during a 6-h hypoglycemic (plasma glucose below 2.8 mmol/l) clamp. Alanine (2 mmol/kg body weight/h) was infused for 3 h to describe the relationship between blood amino nitrogen concentrations and hepatic ureagenesis estimated from urea urine excretion and accumulation in body water. The slope of this relationship is denoted functional hepatic nitrogen clearance (FHNC) and quantifies substrate-independent alterations in hepatic amino nitrogen degradation. In parallel, amino nitrogen balances across muscles were estimated by the forearm flux method. RESULTS: Euglycemia decreased circulating glucagon values (100+/-25 ng/l vs. 160+/-30 ng/l), whereas hypoglycemia doubled glucagon (350+/-45 ng/l, p<0.05). Hepatic nitrogen clearance (FHNC) decreased during hyperinsulinemic euglycemia (19.5+/ 3.4 l/h vs. 30.6+/-5.7 l/h, p<0.01), whereas forearm net uptake of amino nitrogen increased (130+/-40 nmol/100 ml x min vs. control: -10+/-4 nmol/100 ml x min). During hypoglycemia there was a 3-fold increase in hepatic nitrogen clearance up to 83.0+/-16.8 l/h (p<0.01) and increased release of amino nitrogen from the forearm (-100+/-30 nmol/100 ml x min, p<0.01). CONCLUSION: Hypoglycemia in man induces a marked increase in hepatic amino- to urea-N clearance. This catabolic response to hypoglycemia in the liver may be of primary importance for muscle amino acid release. Our data are compatible with the notion that liver and muscle together are responsible for catabolism during hypoglycemia, and that glucagon may be the primary mediator via its effect on liver metabolism. PMID- 10365808 TI - The tumor necrosis factor-alpha promoter correlates with progression of primary biliary cirrhosis. AB - BACKGROUND/AIMS: There have been many studies attempting to identify genes that determine susceptibility to primary biliary cirrhosis (PBC), but few studies have attempted to define the genes that modulate the natural history of the disease. There is a biallelic polymorphism, coined TNF1 and TNF2, in the TNFalpha promoter region at -308. We investigated the relative frequency of the TNF1 and TNF2 alleles in patients with PBC, based on the hypothesis that a polymorphism of the TNFalpha promoter region may be associated with the rate of progression and prognosis of PBC. METHODS: Seventy-one Caucasoid patients with PBC and 133 healthy and unrelated Caucasoid individuals were studied. Genomic DNA was extracted from blood, and the mutation at position -308 of the TNFalpha gene analyzed by PCR and NcoI digestion. RESULTS: In 71 patients with PBC, 56/71 (78.9%) patients were TNF1/TNF1 homozygotes, 14/71 (19.7%) were TNF1/TNF2 heterozygotes and 1/71 (1.4%) were TNF2/TNF2 homozygotes. In 133 healthy individuals, 109/133 (80.5%) patients were TNF1/TNF1 homozygotes, 24/133 (18%) were TNF1/TNF2 heterozygotes. No control individuals were TNF2/TNF2 homozygotes. The difference between the two groups was not statistically significant (p = 0.3684). However, in patients with TNF1/TNF1 the Mayo score for disease severity was 4.596+/-0.157 (mean +/- SEM), compared to 5.637+/-0.420 for patients with TNF1/TNF2. This Mayo score was significantly higher in patients with the TNF1/TNF2 genotype than those with TNF1/TNF1 (p = 0.0140), with an odds ratio of 4.9. CONCLUSIONS: Our data demonstrate that the presence of the TNF2 allele may be associated with a higher Mayo score, and thus with patients in a more advanced clinical stage. These data have both theoretical and clinical implications. PMID- 10365809 TI - Comparison of three doses of ursodeoxycholic acid in the treatment of primary biliary cirrhosis: a randomized trial. AB - BACKGROUND/AIM: Ursodeoxycholic acid in doses of 13-15 mg x kg(-1) x day(-1), is a safe and cost-effective treatment for patients with primary biliary cirrhosis. However, very limited information exists regarding the most appropriate dose of ursodeoxycholic acid. The aim of the study was to compare three dosages of ursodeoxycholic acid with respect to changes in liver biochemistries, Mayo risk score, biliary enrichment with ursodeoxycholic acid and side effects over at least a 1-year period. METHODS: A total of 155 patients were randomized to receive low- (5-7 mg x kg(-1) x day(-1)), standard-(13-15 mg x kg(-1) x day(-1)), and high- (23-25 mg x kg(-1) x day(-1)) doses of ursodeoxycholic acid. RESULTS: The improvements in alkaline phosphatase (p = 0.0001), aspartate aminotransferase (p = 0.0001), Mayo risk score (p = 0.002), and ursodeoxycholic acid enrichment (p = 0.0001) were significantly greater in the standard- and high-dose groups compared to the low-dose group, but not between the standard- and high-dose groups. Changes in serum bilirubin were similar between the three groups (p = 0.07). No significant effects on symptoms were noted with any dose. No patients discontinued ursodeoxycholic acid because of side effects or toxicity. CONCLUSIONS: Ursodeoxycholic acid in doses of 5-25 mg x kg(-1) x day(-1) is safe and well tolerated. The dose of 13-15 mg x kg(-1) x day(-1) appears to be the preferred dose for patients with primary biliary cirrhosis. PMID- 10365810 TI - Gene transfer to the rat biliary tract with the HVJ-cationic liposome method. AB - BACKGROUND/AIMS: The ability to transfer foreign genes into the biliary tract would be useful for the treatment of biliary tract diseases, including cancer, cystic fibrosis and other genetic diseases. To introduce a foreign gene precisely into the rat biliary epithelial cells, we developed a new technique, inserting a polyethylene catheter into the common bile duct through the papilla of Vater by use of a fusigenic cationic liposome with hemagglutinating virus of Japan (HVJ cationic liposome). METHODS: Transfection efficiency was estimated with the use of FITC-oligonucleotides (FITC-ODNs) and cDNA of beta-galactosidase (pCAG-lacZ). RESULTS: FITC-ODNs encapsulated in HVJ-cationic liposome were effectively transfected into cell nuclei of human cholangiocellular carcinoma in vitro after a 30-min incubation as compared with the simple application of naked FITC-ODNs. After in vivo injection of FITC-ODNs using the HVJ-cationic liposome method through the papilla of Vater, fluorescence accumulation was observed only in the epithelial cells of the biliary tract, but not in the parenchymal cells of the liver. Beta-galactosidase expression was observed in the biliary epithelial cells 3 days after the transfection of pCAG-lacZ and was also detected at 14 days, but not at 28 days, without obvious cytotoxicity. CONCLUSIONS: HVJ-cationic liposome mediated gene transfer to the biliary tract via the papilla of Vater is a minimally-invasive and an effective gene-delivery method for site-specific targeting to the epithelial cells of the biliary tract, which could be applied to the treatment of human biliary tract diseases. PMID- 10365811 TI - Effects of amino acids on bile acid-dependent and independent bile flow in the isolated perfused rat liver. AB - BACKGROUND/AIMS: Conflicting data on the effects of amino acids on biliary function led us to investigate their interaction with taurocholate in the perfused rat liver model. METHODS: To investigate the influence of amino acids on the bile acid-independent component of bile flow, 12 livers were perfused with (n = 6) and without (n = 6) amino acid addition from t30 min. For the study of bile acid-dependent bile flow, 24 livers were perfused under 8 experimental conditions according to the perfusate taurocholate concentration (12.5, 25, 37.5 or 50 microM) and whether amino acids were or were not added from t30 min. RESULTS: In the absence of taurocholate, amino acids induced a 40% (p<0.01) decrease in bile flow together with an increase in hepatic water content (17.8%, p< 0.05). Thus, amino acids exert an inhibitory effect on bile acid-independent bile flow despite the postulated cell swelling-dependent increase in bile flow. When livers were perfused at various taurocholate concentrations, amino acids induced, in addition to their inhibitory effect on bile acid-independent bile flow, a significant increase in taurocholate apparent choleretic activity (13.2 microl/micromol vs. 10.6 microl/micromol; p = 0.05), while taurocholate intrinsic clearance was significantly decreased (4.5+/-1.2 ml x min(-1) x g(-1) vs. 6.1+/-1.3 ml x min( 1) x g(-1); p<0.01). CONCLUSIONS: These data suggest that at physiological bile acid concentrations amino acids exert an inhibitory effect on both bile acid dependent and- independent bile flow, whereas at higher taurocholate concentrations this inhibitory effect disappears, probably because of cell swelling-dependent mechanisms. PMID- 10365812 TI - Human and rat hepatic stellate cells produce stem cell factor: a possible mechanism for mast cell recruitment in liver fibrosis. AB - BACKGROUND/AIMS: Mast cell numbers are markedly increased in advanced liver fibrosis. Stem cell factor may recruit mast cells to the liver following injury as it induces mast cell proliferation, survival and differentiation from resident tissue precursors. This study examines stem cell factor production in human fibrotic liver and by hepatic stellate cells during culture in vitro. METHODS: Stem cell factor production was examined in human fibrotic livers by ELISA and in human and rat hepatic stellate cell cultures using reverse transcription polymerase chain reaction (RT-PCR), Northern blotting, Western blotting and immunocytochemistry. Co-culture studies examined adhesion between hepatic stellate cells and purified mast cells. RESULTS: RT-PCR showed stem cell factor mRNA was more consistently expressed in fibrotic human livers relative to normal, and ELISA confirmed this by showing stem cell factor protein was significantly increased 2-3-fold in homogenates of human cirrhotic liver (primary biliary cirrhosis, primary sclerosing cholangitis) relative to normal. RT-PCR detected stem cell factor mRNA in human and rat hepatic stellate cells activated by culture on plastic. This was confirmed by Western blotting, which showed that freshly isolated hepatic stellate cells expressed relatively little 30 kD stem cell factor compared to late primary culture activated hepatic stellate cells (14 day) and passaged hepatic stellate cells. As assessed by fluorescence immunocytochemistry, stem cell factor protein was homogeneously expressed by populations of culture-activated rat hepatic stellate cells. During co-culture, purified human skin mast cells adhered to hepatic stellate cell monolayers on plastic, and this adherence was inhibited >50% by addition of antibodies against stem cell factor. CONCLUSIONS: Hepatic stellate cells activated in vitro produce stem cell factor. These cells may play an important role in recruiting mast cells to liver during injury and fibrosis. PMID- 10365813 TI - Evaluation of role of mast cells in the development of liver fibrosis using mast cell-deficient rats and mice. AB - BACKGROUND/AIMS: Several studies have suggested that mast cells participate in the development of liver fibrosis in rodent models. In this study mast cell deficient mutant Ws/Ws rats and W/Wv mice were used to examine whether mast cells are involved in the development of liver fibrosis. METHODS: Liver fibrosis was induced in rats by bile duct resection (BDR), and by intraperitoneal injections of carbon tetrachloride (CCl4) or porcine serum, and in mice by intragastric administrations of CCl4, and BDR. The degree of fibrosis was evaluated by measuring the hydroxyproline content (microg/mg tissue) of the liver as an index of the collagen content. The density of mast cells (number/cm2 liver section) was determined by counting mast cells in liver sections stained with alcian blue. RESULTS: In the liver of control non-mutant (+/+) rats, mast cells were found principally in portal areas, and their average density was 200-300/cm2 liver section. BDR, and treatments with CCl4 and porcine serum increased the density of mast cells in the liver of +/+ rats several-fold, and induced liver fibrosis, increasing the liver hydroxyproline content markedly. BDR, and treatments with CCl4 and porcine serum also induced liver fibrosis in Ws/Ws rats, increasing the liver hydroxyproline content to a similar or higher level than that in +/+ rats. However, the average densities of mast cells in the liver of Ws/Ws rats after BDR and treatment with CCl4 and porcine serum were at most 10.2/cm2 liver section. The density of mast cells in the liver of control +/+ mice was extremely low (average, less than 2), and neither BDR nor treatment with CCl4 caused any significant increase in their density, whereas these treatments induced liver fibrosis and markedly increased the liver hydroxyproline content. Furthermore, treatment with CCl4 induced fibrosis in the liver of W/Wv mice similarly to that in +/+ mice, but the density of mast cells in the liver of W/Wv mice was very low (average, less than 1), and was not increased by treatment with CCl4. CONCLUSIONS: The present results indicate that mast cells play no role in the development of liver fibrosis in rats and mice. PMID- 10365814 TI - The Na+/H+ exchanger modulates the fibrogenic effect of oxidative stress in rat hepatic stellate cells. AB - BACKGROUND/AIMS: Oxidative stress is associated with liver fibrosis in vivo and with hepatic stellate cell (HSC) activation in vitro, but the intracellular mechanisms mediating these effects are mostly unknown. The Na+/H+ exchanger plays a key role in regulating the cell cycle, and is involved in HSC proliferation. Its role in different HSC features, such as collagen accumulation, is still unknown. We thus evaluated if the Na+/H+ exchanger modulates the fibrogenic effect of oxidative stress in rat HSC. METHODS: HSC were incubated with 0.1 mM ferric nitrilotriacetate complex (FeNTA). Intracellular hydroperoxides and malonildialdehyde (MDA) levels in the culture media were measured by the dichlorofluorescein and TBARS method, respectively. Intracellular pH and Na+/H+ exchanger activity were measured using the fluorescent dye BCECF. Cell proliferation was measured by immunohistochemistry for bromodeoxyuridine incorporation. Collagen type I accumulation in the culture media was measured by ELISA. RESULTS: HSC incubation with FeNTA resulted in a significant production of intracellular hydroperoxides and MDA, associated with increased Na+/H+ exchange activity and baseline intracellular pH (pHi). Exposure of HSC to FeNTA significantly enhanced the number of proliferating HSC and collagen type I levels in the culture medium. All these effects were reversed by the antioxidant resveratrol and by the Na+/H+ exchanger inhibitor amiloride. CONCLUSIONS: This study indicates that the Na+/H+ exchanger might represent a common mediator of the different effects induced by oxidative stress on HSC. The reduction in cell proliferation and collagen synthesis induced by amiloride could represent a new therapeutic challenge in liver fibrosis. PMID- 10365815 TI - Hepatic arterial pulsatility index in cirrhosis: correlation with portal pressure. AB - BACKGROUND/AIMS: Determination of the pulsatility index by means of duplex sonography provides the opportunity to evaluate the vascular resistance of the hepatic artery noninvasively. The aim of this study was to investigate the relationship between the hepatic arterial pulsatility index and the hepatic venous pressure gradient in cirrhosis. METHODS: In 50 patients with cirrhosis, hepatic venous pressure gradient was determined in the fasting state. Immediately thereafter, hepatic arterial pulsatility index and portal blood flow velocity were measured by duplex sonography with no knowledge of hepatic venous pressure values. In addition, the duplex parameters were determined in 20 controls. RESULTS: Hepatic arterial pulsatility index was significantly higher in patients with cirrhosis than in controls (0.92+/-0.1 vs. 1.14+/-0.18; p<0.001) and directly correlated with the hepatic venous pressure gradient (r = 0.7; p<0.001). Furthermore, weak correlations were found between hepatic arterial pulsatility index and Child-Pugh score (r = 0.49; p<0.01) and between portal blood flow velocity and hepatic venous pressure gradient (r = -0.48; p<0.01). CONCLUSION: In cirrhosis the hepatic arterial vascular resistance seems to increase parallel to the rise of the portal pressure. Therefore, duplex sonographic determination of the hepatic arterial pulsatility index may contribute to the noninvasive evaluation of portal hypertension. PMID- 10365816 TI - Hepatopulmonary syndrome associated with cardiorespiratory disease. AB - BACKGROUND/AIMS: Hepatopulmonary syndrome is defined as a clinical triad including chronic liver disease, abnormal pulmonary gas exchange resulting ultimately in profound arterial hypoxaemia, and evidence of intrapulmonary vascular dilatations. We report five patients with liver cirrhosis diagnosed with hepatopulmonary syndrome who had associated chronic obstructive or restrictive respiratory diseases. METHODS: Clinical, radiographic and constrast-enhanced echocardiographic findings, and systemic and pulmonary haemodynamic and gas exchange, including ventilation-perfusion distributions, measurements were assessed in all five patients. RESULTS: Echocardiography was consistent with the presence of intrapulmonary vasodilation without intracardiac abnormalities, and high resolution computed tomographic scan features were compatible with clinical (3 cases) or histopathological diagnoses (2 cases) of associated respiratory disorders. The most common prominent functional findings were moderate to severe arterial hypoxaemia, caused by moderately to severely increased intrapulmonary shunting and/or mild to moderate low ventilation-perfusion areas, and hypocarbia along with an increased cardiac output and a low pulmonary artery pressure and vascular resistance. CONCLUSIONS: These functional characteristics, classically reported in the setting of clinically stable, uncomplicated hepatopulmonary syndrome, conform to a distinctively unique, chronic gas exchange pattern. Equally important, these pulmonary haemodynamic-gas exchange hallmarks are not influenced by the co-existence of chronic cardiorespiratory disease states. These data may have clinical relevance for elective indication of hepatic transplantation in patients with life-threatening hepatopulmonary syndrome. PMID- 10365817 TI - Prognostic significance of hepatic encephalopathy in patients with cirrhosis. AB - BACKGROUND: There are numerous studies concerning the natural history and prognostic factors in cirrhosis, the results of which are useful in selecting liver transplant candidates. However, little attention has been paid to the prognostic significance of hepatic encephalopathy despite the high frequency of this complication. METHODS: We reviewed the charts of 111 cirrhotic patients who developed a first episode of acute hepatic encephalopathy to determine their survival probability and to identify prognostic factors. RESULTS: During follow up (12+/-17 months), 82 (74%) patients died. The survival probability was 42% at 1 year of follow-up and 23% at 3 years. With univariate analyses followed by a multivariate analysis, 7 out of 30 clinical and standard laboratory variables were significantly associated with poor prognosis: male sex, increased serum bilirubin, alkaline phosphatase, potassium and blood urea nitrogen, and decreased serum albumin and prothrombin activity. Patients were classified into two groups according to a prognostic index calculated from these 7 variables. Survival probability at 1 and 3 years was 73% and 38%, respectively, in patients with a low prognostic index, and 10% and 3% in patients with a high prognostic index. CONCLUSION: Hepatic encephalopathy is associated with short survival in cirrhotic patients. Although these patients can be classified into several groups with a different prognosis, the survival probability in every group is lower than that currently expected after liver transplantation. Therefore, cirrhotic patients developing a first episode of acute hepatic encephalopathy should be considered as potential candidates for this therapeutic procedure. PMID- 10365818 TI - Vascular nitric oxide production during the development of two experimental models of portal hypertension. AB - BACKGROUND/AIMS: The aim of this study was to determine the relative roles of constitutive NOS (NOS3) and inducible NOS (NOS2) isoforms during the development of two models of portal hypertension in rats. METHODS: Vascular reactivity of aortic rings for norepinephrine was performed in control, sham-operated, portal vein-stenosed and secondary biliary cirrhotic rats 1, 4, 7, 14 and 28 days after surgery. NOS activity and nitrate plasma levels were also measured. RESULTS: An impaired response to norepinephrine observed in sham-operated, portal-vein stenosed and cirrhotic rats at days 1 and 4 compared with controls was reversed after L-NNA and aminoguanidine. Portal hypertensive rats remained hyporeactive at days 7, 14 and 28 compared with sham-operated rats. At days 7 and 14 in portal vein-stenosed rats, vascular hyporeactivity was reversed by L-NNA and W7. At days 14 and 28 in cirrhotic rats, vascular hyporeactivity was reversed by L-NNA and W7. Nitrate levels increased at day 1 in the 3 groups, and increased at days 14 and 28 in portal hypertensive rats. Total NOS-activity increased in cirrhotic rats at day 28, in portal-vein-stenosed rats at day 14, and in sham-operated rats at day 1 compared to controls. NOS2 activity increased only in sham-operated rats at day 1. CONCLUSIONS: This study shows that for two models of portal hypertension, increased NO production in the first days is related to NOS2 induction secondary to surgery. On the other hand, when portal hypertension has fully developed, the NOS3 isoform appears to play the major role. PMID- 10365819 TI - Effect of simvastatin, a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, on alpha-fetoprotein gene expression through interaction with the ras mediated pathway. AB - BACKGROUND/AIMS: The ras proto-oncogene encodes a small GTP-binding protein (Ras) which regulates cell growth and differentiation by relaying signals from the cell surface to the nucleus. In the present study, the role of Ras signal transduction pathway in alpha-fetoprotein (AFP) gene expression was evaluated in HuH-7 human hepatoma cells using simvastatin, a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, which blocks Ras function through inhibition of farnesylation, and the ras(val-12) expression vector. METHODS: The HuH-7 cells were treated with simvastatin (10 micromol/l), or both simvastatin and mevalonate (300 micromol/l), and numbers of viable cells were counted after treatment. To elucidate the effects of simvastatin on AFP gene expression and the interactive effect of simvastatin on Ras signal transduction pathway, Northern blotting and transient chloramphenicol acetyltransferase plasmid transfection assays were performed. RESULTS: Cell growth was inhibited by simvastatin, and this growth inhibition was restored by addition of mevalonate. Levels of AFP mRNA but not albumin mRNA were elevated by simvastatin in a dose-dependent manner (1-10 micromol/l). AFP promoter and enhancer activities were stimulated by simvastatin. In contrast, both activities were repressed by transfection with the ras(val-12) expression vector. The ras(val-12)-mediated repression was restored by simvastatin and returned to the repressed level by simvastatin plus mevalonate. CONCLUSIONS: These results indicate that the Ras signal transduction pathway functions to down-regulate the AFP gene transcription in human hepatoma cells. PMID- 10365820 TI - Effect of vascular endothelial growth factor on hepatic regenerative activity following partial hepatectomy in rats. AB - BACKGROUND/AIMS: Vascular endothelial growth factor (VEGF) is an angiogenic factor with a growth-promoting effect that is thought to be restricted to vascular endothelial cells. Its essential role during liver regeneration has yet to be determined. The aim of this study was to document the effect of exogenous VEGF administration on liver regeneration in rats undergoing submaximal hepatic resections. METHODS: Adult male Sprague-Dawley rats (n = 4/group) undergoing 30% partial hepatectomy were administered 200 ng VEGF165 intravenously and were sacrificed at 24, 36, and 48 h postoperatively. Liver regeneration was monitored by measuring the restituted liver mass, proliferating cell nuclear antigen (PCNA) immunostaining, and hepatic PCNA protein by Western blot. RESULTS: Changes in restituted liver mass 48 h postsurgery were more prominent, but did not differ statistically between VEGF-treated and control rats (47% vs. 29%; p<0.06). Nevertheless, PCNA immunostaining showed increased labeling index of hepatocytes, apparent at 36 and 48 h after partial hepatectomy (38% vs. 18% [p<0.041 and 42% vs. 11% [p<0.021], respectively). Hepatic PCNA proteins measured by Western blot showed a 3-fold increase in VEGF-treated rats 48 h postsurgery compared with controls (p<0.01). CONCLUSION: Exogenous VEGF administration early after partial hepatectomy stimulates liver regeneration in rats. Whether or not VEGF165 is a direct mitogen for hepatocytes remains to be determined. PMID- 10365821 TI - Hepatocyte growth factor activates the AP-1 complex: a comparison between normal and transformed rat hepatocytes. AB - BACKGROUND/AIMS: Stimulation of activator protein-1 (AP-1), a Fos/Jun complex, is a key event in the cell response to growth factors. We have investigated whether hepatocyte growth factor (HGF) induces differential AP-1 responses in normal and transformed rat hepatocytes, the 7777 cells. METHODS: Primocultures of isolated hepatocytes or 7777 cells were stimulated with HGF. Gene expression was evaluated by ribonuclease protection assay and Western blot analysis. AP-1 DNA binding activity was measured by electrophoretic mobility shift assay. Identification of the proteins bound to the probes was made by supershift assays with specific antibodies. Cells were electroporated with plasmids containing an AP-1-dependent chloramphenicol acetyl transferase (CAT) gene, and CAT activity was measured 24 h after treatment with medium alone or HGF. RESULTS: In both cell types, HGF triggered the same program of jun family mRNA activation, but distinct Fos/Jun proteins accumulated in the nucleus. HGF increased DNA-binding activity to the phorbol 12-O-tetradecanoate-13-acetate responsive element (TRE) in both cell types, but distinct TRE-binding proteins were recruited in the AP-1 dimers. HGF also increased consistently binding to a cAMP responsive element (CRE) in hepatocytes only. Finally, HGF triggered TRE- and CRE-dependent gene activations in hepatocytes but TRE-dependent gene activation alone in 7777 cells. CONCLUSIONS: HGF-induced AP-1 activation leads to the formation of distinct dimers with different functional capacities in normal and transformed hepatocytes. These data suggest the importance of qualitative abnormalities of the AP-1 complex for the establishment or maintainance of a transformed phenotype. PMID- 10365822 TI - Carbonyl-iron supplementation induces hepatocyte nuclear changes in BALB/CJ male mice. AB - BACKGROUND/AIMS: In humans, chronic iron excess may induce hepatic fibrosis and/or hepatocellular carcinoma. This work was undertaken to investigate hepatic iron overload outcome in iron-overloaded mice. METHODS: BALB/cJ male mice received supplements of 0, 0.5, 1.5 and 3% carbonyl-iron for 2, 4, 8 and 12 months. Histological staining, immunohistochemistry using ferritin antibodies and electron microscopic studies were performed on liver. Liver iron concentration was measured biochemically. Mitotic index and hepatocyte nuclear size were evaluated on Feulgen-stained slides. RESULTS: Liver iron concentration was increased, reaching 13 times control value after 12 months in 3% iron-overloaded mice, and iron was found predominantly in hepatocytes, with a porto-centrolobular decreasing gradient. Neither hepatic fibrosis nor hepatocellular carcinoma was found. Perls' stain positive inclusions containing ferritin were found within hepatocyte nuclei in 3%-overloaded mice. Electron microscopy disclosed that inclusions consisted of ferritin particle aggregates without a limiting membrane. Mice overloaded with 3% iron for 12 months showed larger hepatocyte nuclei than control mice and a mitotic index increase with presence of abnormal tripolar mitotic figures. In addition, some iron-free hepatocytes were observed. CONCLUSIONS: Carbonyl-iron supplementation produces significant iron overload in mice but does not result in liver fibrosis or hepatocellular carcinoma after 12 months. However, nuclear changes were produced in hepatocytes, and occasional iron-free hepatocytes were observed: these may represent preneoplastic changes caused by iron overload. PMID- 10365823 TI - Increased expression of interleukin-4 during liver allograft rejection. AB - BACKGROUND/AIMS: To investigate the respective roles of interleukin-2 (IL-2) and IL-4 during rejection, we evaluated the expression of IL-2, IL-2 receptor and IL 4 in human liver allografts. METHODS: Immunohistochemistry and RT-PCR were performed in liver biopsies. To determine the effects of immunosuppression and cholestasis in IL-4 production, in vitro experiments were also designed. RESULTS: IL-2 protein and its mRNA were absent in the liver, with minimal expression of IL 2 receptor, during rejection. In contrast, IL-4 protein and its mRNA were highly expressed during acute and chronic rejection, whereas this expression was absent in stable liver transplant recipients. In vitro, cyclosporine potently inhibited IL-2 and IL-2 receptor expression of activated mononuclear blood cells, but poorly inhibited IL-4 expression. Chenodeoxycholic acid decreased IL-2 and IL-2 receptor expression, but increased IL-4 expression. CONCLUSIONS: During liver allograft rejection, IL-2 pathway is down-regulated, while IL-4 expression is increased by cholestasis and poorly inhibited by cyclosporine. These data suggest that IL-4 is involved in the mechanisms of liver allograft rejection in patients treated with cyclosporine. PMID- 10365824 TI - An enhancement of nitric oxide production regulates energy metabolism in rat hepatocytes after a partial hepatectomy. AB - BACKGROUND/AIMS: Infection after a liver resection often results in hepatic failure. Nitric oxide is one of the candidates which has been suspected to cause cellular dysfunction during infection in the liver. We have previously reported that the inflammatory cytokine interleukin-1beta (IL-1beta) induced the expression of the inducible nitric oxide synthase gene in primary cultured rat hepatocytes. We hypothesized that an enhancement of nitric oxide production after the resection was implicated in a change in liver energy metabolism, thus resulting in liver dysfunction. METHODS: In this study, we performed a 70% hepatectomy or a sham operation in rats, and then isolated hepatocytes from the remnant liver by collagenase perfusion. The cultured hepatocytes were treated with cytokines including IL-1beta. The effects on nitric oxide induction, the ATP content and ketone body ratio (acetoacetate/beta-hydroxybutyrate) were then compared between the partial hepatectomized (PH) and sham-operated (control) rats. RESULTS: IL-1beta augmented the induction of nitric oxide production two fold in hepatocytes from the PH rats as compared to the control rats. IL-1beta markedly decreased the ATP content in the PH rats, although IL-1beta also decreased the ATP content in the control rats, but to a lesser extent. IL-1beta also decreased the ketone body ratio in both groups. The addition of L-arginine further stimulated the inhibition of the ATP levels and the ketone body ratio concomitantly with increased nitric oxide production in the PH rats. N(G) monomethyl-L-arginine, an inhibitor of nitric oxide synthase, abolished the effects of IL-1beta on the ATP levels and ketone body ratio, as well as on the nitric oxide production. CONCLUSIONS: These results demonstrate that the decreased ATP content observed in PH rats resulted from an increase in nitric oxide production. The decrease in ketone body ratio indicates that nitric oxide induced mitochondrial dysfunction contributes significantly to ATP attenuation in hepatocytes. Therefore, the regulation of nitric oxide induction may be crucial for preventing liver failure after a hepatic resection. PMID- 10365825 TI - Progressive development of diffuse liver hemangiomatosis. AB - Diffuse liver hemangiomatosis is extremely rare. The etiology and natural history of the disease are unknown. It is also unclear whether tumor growth is induced or modulated by drug therapy. Tumor recurrence after ablative therapy has not been described in patients with diffuse liver hemangiomatosis. Diffuse hemangiomatosis of the left hepatic lobe was suspected in a 35-year-old woman by ultrasonography, CT and hepatic arteriography, and confirmed by laparotomy and biopsies. The patient denied any drug or estrogen use. The tumor was removed by left hepatectomy. Two and six years later, the patient was again hospitalized with progressive tumor growth into the right hepatic lobe. Although diffuse liver hemangiomatosis is a rare disease, its diagnosis should be considered in patients with progressive tumor growth in one or both hepatic lobes. The absence of drug intake or estrogen use does not exclude the diagnosis. PMID- 10365826 TI - Nodule in nodule: malignant transformation of a macroregenerative nodule in cirrhosis revealed by duplex-Doppler. PMID- 10365827 TI - EASL International Consensus Conference on Hepatitis C. Paris, 26-28, February 1999, Consensus Statement. European Association for the Study of the Liver. PMID- 10365828 TI - Outcome of chronic hepatitis B in children with normal transaminase levels. PMID- 10365829 TI - Prognosis of human pancreatic adenocarcinoma: review of clinical and histopathological variables and possible uses of new molecular methods. AB - Cancer of the exocrine pancreas is a disease of considerable importance in gastroenterology. In Western countries it is the fourth commonest cause of death from cancer after those of lung, colorectal, and breast. The incidence of pancreatic carcinoma has increased in Northern Europe and North America during recent decades and contrary to for example, lung, gastric and oesophageal carcinoma, its incidence is still increasing the annual incidence is about 8 10/100,000 population. The causes of its increased incidence are unknown, as is the aetiology of the disease itself. Pancreatic cancer generally grows without symptoms until late in its natural history and there are therefore many discouraging unresolved problems in management. However, some progress has been made in understanding the molecular basis of pancreatic carcinogenesis. Recent molecular pathological studies have described mutation or overexpression of important oncogenes such as K-ras and bcl-2 and deletions of tumour suppression genes such p53, DPC4, CDKN2, and the Rb gene. The present prognosis of pancreatic cancer is, however, controversial, and as these new markers may have the potential for improving our ability to predict its course, we have reviewed current knowledge, and concentrated on the classic and the recently-introduced factors in the prediction of its prognosis. PMID- 10365830 TI - Association of low preoperative serum albumin concentrations and the acute phase response. AB - OBJECTIVE: To investigate the incidence of a preoperative acute phase response and its association with low albumin concentrations. DESIGN: Prospective open study. SETTING: Teaching hospital, Germany. SUBJECTS: 225 patients who were to undergo major abdominal operations, and who had no acute infections. INTERVENTIONS: Measurements of serum concentrations of albumin, C-reactive protein (CRP), alpha-1 antitrypsin, and interleukin-6 (IL-6). RESULTS: Abnormal concentrations of acute phase proteins (indicating an acute phase response) were detected in 43 of 225 patients (19%). The mean (SD) albumin concentration in these patients (35[5]g/L) was lower than that of patients who did not mount an acute phase response preoperatively (40[5]g/L). High concentrations of CRP (> or =60mg/L) were associated with low albumin concentrations (33[5]g/L); high alpha-1 antitrypsin concentrations (> or =4.0g/L) were associated with low albumin concentrations (34[6]g/L); and high IL-6 concentrations (> or =4pg/ml) were associated with low albumin concentrations (37[6]g/L) compared with a mean(SD) albumin concentration of 40(5)g/L in patients who had no evidence of an acute phase response. CONCLUSION: A metabolic response to disease referred to as an acute phase response may explain low preoperative albumin concentrations. This association interferes with the association of low preoperative albumin concentrations and malnutrition. It is a new aspect of preoperative risk evaluation and may indicate a potential for prevention. PMID- 10365831 TI - Reconstruction of the supra-aortic trunks. AB - OBJECTIVE: To review our 10-year experience of reconstruction of the supra-aortic trunks. DESIGN: Retrospective study. SETTING: Teaching hospital, The Netherlands. SUBJECTS: 47 patients who required reconstruction of the supra-aortic trunks for stenotic or occlusive disease between April 1987 and May 1997. INTERVENTIONS: Right-sided bifurcation graft through a sternotomy (n = 25), left-sided thoracotomy (n = 1), and extra-anatomic bypass (n = 21). MAIN OUTCOME MEASURES: Morbidity, mortality, and long term patency. RESULTS: 3 patients died (6%); 7 (15%) developed major complications (leak from the brachiocephalic stump, n = 2, and acute occlusion of the bypass graft, n = 5) all of which were successfully treated by immediate reoperation; and 9 (19%) developed minor complications, all of which resolved within 3 months. The median follow up was 36 months (range 1 108), and the 3-year patency rate was 80%. No patient died during the follow up period, but a further 3 were lost to follow up. The remaining 41 were all assessed by duplex scanning or angiography, and 3 required further operation for recurrent symptoms; 33 remained completely free of symptoms. CONCLUSION: Symptomatic stenotic or occlusive lesions of the supra-aortic trunks can be treated with acceptable morbidity and mortality, giving long term benefit to patients. PMID- 10365832 TI - Clinicopathological features of early gastric cancer: results of 100 cases from a rural general hospital. AB - OBJECTIVE: To evaluate 100 patients with early gastric cancer from the point of view of early detection, clinicopathological variables, and long term results. DESIGN: Retrospective study. SETTING: Rural general hospital, Japan. SUBJECTS: 100 patients with early gastric cancer (confined to the epithelium, lamina propria, or submucosa) out of a total of 197 who had gastric cancers resected for cure between May 1986 and April 1996. INTERVENTIONS: Subtotal gastrectomy (n = 87), total gastrectomy (n = 8), proximal gastrectomy (n = 2), and local wedge resection (n = 3). MAIN OUTCOME MEASURES: Histopathological features and outcome. RESULTS: The mean annual incidence of early gastric cancer was 51% (range 35% 70%). 16/59 patients with mucosal cancer (37%) and 18/41 with submucosal cancer (44%) presented with symptoms of the disease. The diagnosis was made in 62 by endoscopy, and in only 2 by upper gastrointestinal radiographic examination. None of the 59 with mucosal cancer had lymphatic invasion, and only 1 had a lymph node metastasis. Among the 41 with submucosal cancer, however, 15 had lymphatic invasion (37%), 13 had venous invasion (32%), and 2 had lymph node metastases (5%). 83 patients were alive with no sign of recurrence at the time of writing (median follow up 62 months, range 12-136). One patient with a tumour that produced alpha-fetoprotein died of hepatic metastases 23 months after subtotal gastrectomy. 9 patients developed second cancers, and 6 died of these with no signs of recurrence of early gastric cancer. The overall 5 and 10 year survival rates were 82% and 66%, and the corresponding disease-specific survival rates for 85 patients were both 98%. CONCLUSIONS: Excellent long term results can be achieved in the treatment of early gastric cancer, even in a non-specialist centre. Patients with early gastric cancer should have their alpha-fetoprotein concentration measured, and be examined for the presence of other malignant disease both before and after treatment of the gastric cancer. PMID- 10365833 TI - Is elective hernia repair worthwhile in old patients? AB - OBJECTIVE: To find out if elective herniorraphy in patients aged 75 and over is worthwhile. DESIGN: Retrospective study. SETTING: District hospital, Sweden. SUBJECTS: 146 consecutive patients aged 75 years or more, who had their hernias repaired during the period 1992-95. MAIN OUTCOME MEASURES: Patient satisfaction measured by a five-point analogue scale. Clinical and personal details, morbidity, mortality, and surgical variables were obtained from case records. RESULTS: Community social service was not required by 114 (78%) of the patients and 15 (22%) had no preoperative complaints. Our patients rated their satisfaction with their choice to have an operation, as well as its effect on their preoperative symptoms as 4.9. Emergency operations (p = 0.02), femoral hernias (p = 0.01) and direct inguinal hernias (direct:indirect ratio 0.81) were more common in this age group. Femoral and direct inguinal hernias tended to recur more often than usual. Emergency operation, dementia, and diabetes were associated with a reduced short-term survival. CONCLUSION: Elective hernia repair in an elderly population is highly appreciated by the patients, and worthwhile. If coexisting disease and domestic arrangements are controlled, the patients' need for hospital care can be minimised. Mesh is recommended in femoral and direct inguinal hernias, which were associated with an increased reoperation frequency. A more vigilant protocol of indications for hernia surgery in the aged may minimise the need for both emergency and unnecessary operations. PMID- 10365834 TI - Comparison of polydioxanone (PDS) and polypropylene (Prolene) for Shouldice repair of primary inguinal hernias: a prospective randomised trial. AB - OBJECTIVE: To compare the recurrence rates after Shouldice operation for primary inguinal hernias, using non-absorbable polypropylene or absorbable polydioxanone suture material. DESIGN: Randomised, controlled trial. SETTING: Teaching hospital, Germany. SUBJECTS: 220 male patients who had 233 elective Shouldice repairs of primary inguinal hernias, 201 of whom were followed up. INTERVENTION: Standard Shouldice procedure with 2/0 polypropylene (Prolene, n = 98) or 2/0 polydioxanone (PDS, n = 103). MAIN OUTCOME MEASURES: Recurrence rates after a minimum follow-up of 24 months (range 24-48, mean 31). RESULTS: Numbers of early complications were similar in the two groups; there were 2 wound infections in each. A total of 193 patients with 201 repairs had a documented follow-up (86%). There were 6 recurrences in the PDS group and 5 in the Prolene group, giving a total recurrence rate of 5%. This difference was not significant (Fisher's exact test, p = 1.0). CONCLUSION: Recurrence rates in both groups were higher than expected, but there was no difference between the two groups. PMID- 10365835 TI - Incidence and aetiological factors in pilonidal sinus among Turkish soldiers. AB - OBJECTIVE: To study the incidence and causes of pilonidal sinus in Turkish soldiers. DESIGN: Open study by questionnaire. SETTING: Military and University hospitals, Turkey. SUBJECTS: 1000 soldiers who presented for their first medical examination. MAIN OUTCOME MEASURES: Correlation between factors known to be associated with pilonidal sinus, and incidence of pilonidal sinus. RESULTS: 88/1000 soldiers had pilonidal sinuses; in 48 they were symptomatic and in 40 asymptomatic. The factors associated with the presence of a pilonidal sinus were: family history of pilonidal sinus (18/88 compared with 32/912, p < 0.0001); obesity defined as weight over 90 kg (34/88 compared with 32/912, p < 0.0001); being the driver of a vehicle (58/88 compared with 308/912, p < 0.0001); and the incidence of folliculitis or a furuncle at another site on the body (22/88 compared with 64/912, p < 0.0001). CONCLUSIONS: Pilonidal sinus is an acquired condition, penetration of hair is the main cause, and the disease can be prevented if the aetiological factors are understood. PMID- 10365836 TI - Prospective study of parathyroid graft function in patients with renal hyperparathyroidism after total parathyroidectomy and heterotopic autotransplantation by measurement of the intact parathyroid hormone concentrations in both antecubital veins. AB - OBJECTIVE: Evaluation of the value of gradients for intact parathyroid hormone after total parathyroidectomy and heterotopic autotransplantation for renal hyperparathyroidism. DESIGN: Prospective long-term follow-up study. SETTING: Teaching hospital, Germany. SUBJECTS: A total of 115 patients operated on for renal hyperparathyroidism between 1 August 1987 to 15 August 1997. INTERVENTIONS: 100/115 had total parathyroidectomy with autotransplantation. MAIN OUTCOME MEASURES: Analyses of serum calcium, alkaline phosphatase, and intact parathormone in serum 1, 4, 8, 12, 18 and 24 months postoperatively and annually thereafter. Parathormone gradients were calculated as the ratio of the parathormone concentrations in the antecubital venous blood of the grafted and the non-grafted arm. RESULTS: During follow-up (mean 32 months, range 1 month to 9 years), 111 of the 115 patients had one to 10 re-examinations (mean: 4) and in the patients who had had total parathyroidectomy with autotransplantation a total of 437 gradients could be calculated, 91% of which were < or =20. Postoperative hypocalcaemia caused by calcium deficiency of the skeleton led to an increase in parathormone secretion and gradients. Increasing parathormone gradients during follow-up as a result of excessive parathormone secretion in the grafted-arm indicated graft-dependent recurrence. In 6 of the 9 patients with graft-dependent recurrences the gradients exceeded 20. CONCLUSION: The combined sequential assessment of gradients for intact parathyroid hormone and of serum calcium concentrations permits objective evaluation of parathyroid graft function. PMID- 10365837 TI - Endoscopic palliation of oesophageal cancer: results of a prospective comparison of Nd:YAG laser and ethanol injection. AB - OBJECTIVE: To evaluate the effectiveness of intratumoral alcohol injection compared with Nd:YAG laser in the treatment of unresectable fungating cancers of the oesophagus. DESIGN: Prospective, randomised clinical study. SETTING: University hospital, Italy. SUBJECTS AND INTERVENTIONS: 47 consecutive patients were randomly allocated to have endoscopic Nd:YAG laser treatment (n = 24), or intratumoural injection of 98% alcohol (n = 23). MAIN OUTCOME MEASURES: Morbidity, mortality, dysphagia score, survival. RESULTS: One patient in the laser group needed analgesic support during and after the treatment, whereas 18 (78%) of those treated with alcohol experienced mild pain and most of them required analgesics. An improvement of at least 2 points in the dysphagia score was noted in 21 patients (88%) in the laser group and in 18 in the alcohol group (78%). The mean dysphagia-free intervals between each treatment were 30 and 37 days, respectively. The median survival was 6 months in each group. There were no significant differences in the mean dysphagia scores of patients still alive. There were no complications in the laser group, but one oesophageal perforation occurred during the preliminary dilatation before the second session of alcohol injection. There were no procedure-related deaths. CONCLUSION: The two techniques allowed similar palliation of dysphagia and improvement of quality of life. Intratumoral injection of alcohol is an effective and inexpensive therapeutic option in the palliation of fungating oesophageal lesions. PMID- 10365838 TI - Pancreaticogastrostomy compared with pancreaticojejunostomy after pancreaticoduodenectomy. AB - OBJECTIVE: To assess the safety of the pancreatic anastomosis after pancreatico duodenectomy (PD). DESIGN: Non-randomized prospective trial in consecutive patients. SETTING: University hospital. SUBJECTS: 171 consecutive patients with resectable periampullary cancer (80%) or intractable pain due to chronic pancreatitis (20%) undergoing PD. INTERVENTIONS: Pancreaticojejunostomy (PJ) and pancreaticogastrostomy (PG). MAIN OUTCOME MEASURES: Mortality and morbidity rates due to anastomotic leak following PJ and PG. RESULTS: 91 PJ and 80 PG patients were comparable for age, gender, total bilirubin, ASA grading, indication for PD, operating time, pancreas texture, blood loss and replacement. The rate of pancreatic fistula was significantly higher in PJ patients (13%) than in PG patients (3.7%) (12 vs. 3, p = 0.029). Overall death rate was significantly higher after PJ (12%) than after PG (3.7%) (11 vs. 3, p = 0.047). Fatal outcome due to pancreatic leak (3 vs. 1, p = 0.83) and other death rates (8 vs. 2, p = 0.14) were not significantly different in PJ and PG groups, respectively. CONCLUSION: PJ was associated with significantly higher pancreatic leak rate than PG. However, there was no statistically significant difference in mortality rates directly related to pancreatic leak. PMID- 10365839 TI - Correlation between DNA content and p53 deletion in colorectal cancer. AB - OBJECTIVE: To find out whether tumour DNA content correlates with allelic loss of p53 and other pathological features in primary colorectal carcinomas. DESIGN: Ongoing prospective study. SETTING: University hospital, Italy. SUBJECTS: 128 patients who had undergone radical resections for colorectal carcinoma. INTERVENTIONS: Flow cytometric measurement of tumour DNA content and detection of allelic loss on the short arm of chromosome 17 by Southern blot (restriction fragment length polymorphism) analysis in fresh tumour specimens. MAIN OUTCOME MEASURES: Correlation between DNA ploidy and deletion of p53, as well as between these two genetic events and clinicopathological variables. RESULTS: Interpretable DNA histograms were obtained for 122 tumour specimens. Forty-three tumours (35%) were diploid and 79 (65%) aneuploid. The diploid tumours were significantly more common in the proximal colon (from the caecum to the splenic flexure) than in the distal colon (from the descending colon to the rectum) (p = 0.002). The allelic state on the short arm of chromosome 17 was evaluated in 80 heterozygous patients. Forty-four tumour specimens (55%) showed deletion of 17p. Allelic loss of p53 was significantly more common in the distal and rectal tumours than in the proximal ones (p < 0.0001). Aneuploidy was more common among those tumours which had shown deletion of p53 than in those that had not (p = 0.0008). CONCLUSIONS: DNA aneuploidy was significantly associated with the deletion of the p53 gene. This suggests that the functional loss of p53 may favour the growth and establishment of an aneuploid cell population within tumours. Tumours of the proximal and distal colon differ in their genetic nature. PMID- 10365840 TI - Effects of 5-fluorouracil and zinc on healing of colonic anastomoses in rabbits. AB - OBJECTIVE: To find out if 5-fluorouracil (5-FU) given intraperitoneally to rabbits impaired the healing of colonic anastomoses, and whether giving zinc might reverse the effect. DESIGN: Laboratory study. SETTING: Teaching hospital, Turkey. ANIMALS: 32 New Zealand white rabbits. INTERVENTIONS: All animals had 1cm of large bowel resected 10cm proximal to the peritoneal reflection and continuity restored by end-to-end anastomosis. They were divided into four groups and given intraperitoneal injections of saline (control group), 5-FU 10mg/kg/day in a concentration of 5mg/ml saline (5-FU alone group), zinc 2mg/kg/day (zinc alone group), and the same doses of 5-FU and zinc (5-FU + zinc group). The injections were given immediately after operation and daily for 4 days. The rabbits were killed at 7 days. MAIN OUTCOME MEASURES: Bursting pressures, tissue hydroxyproline concentrations, tissue zinc concentrations, and light and electron microscopic appearances. RESULTS: Six rabbits died of the complications of anaesthesia and 4 of sepsis leaving 7, 6, 7, and 6 rats in the four groups respectively. Mean (SD) anastomotic bursting pressures were significantly reduced in the 5-FU group compared with controls (5 (2) compared with 7 (1) mm Hg, p: 0.05) and collagen synthesis (indicated by reduced tissue hydroxyproline concentrations) was also decreased (7.1 (0.9) compared with 9.1 (1.5), p < 0.05). Rabbits given 5-FU + zinc had significantly higher bursting pressures than those given 5-FU alone (9 (2) compared with 5 (2), p: 0.01). Bursting pressures were also significantly higher in those given zinc alone, but hydroxyproline concentrations were similar to those in the control group. Histological examination showed that 5-FU alone significantly impaired the healing process, and those in the 5-FU + zinc group healed better than those in the 5-FU alone group. CONCLUSIONS: 5-FU given intraperitoneally significantly impaired the healing of colonic anastomoses in rabbits, and zinc reversed this effect. PMID- 10365841 TI - Effect of intraperitoneal and extraperitoneal insertion of mesh on bacterial translocation: does it make a difference? AB - OBJECTIVE: To asses the effect of insertion of mesh, with or without contact with the peritoneum, on the induction of bacterial translocation. DESIGN: Open experimental study. SETTING: Surgical research laboratory, Turkey. SUBJECTS: 158 Swiss albino mice. INTERVENTIONS: A defect in the abdominal wall was created. In the control group, the defect was closed primarily. In the extraperitoneal group, polypropylene mesh was sutured over the abdominal wall after primary closure of the peritoneum and in the intraperitoneal group, polypropylene mesh was sutured to close the created defect so that it was in contact with the intestines. MAIN OUTCOME MEASURES: Bacterial translocation at 4, 24 and 48 hours. RESULTS: Insertion of mesh in contact with the peritoneum led to increased bacterial translocation to mesenteric lymph nodes at 4 (p = 0.02) and 48 (p = 0.03) hours compared with insertion without contact. CONCLUSION: Contact between a foreign body and the peritoneum is required to induce bacterial translocation. PMID- 10365842 TI - Small bowel ischaemia-reperfusion increases plasma concentrations of oxidised proteins in rats. AB - OBJECTIVES: To find out whether plasma concentrations of protein carbonyl (a specific marker of oxidative damage of proteins) are increased during intestinal ischaemia-reperfusion and whether they are correlated with von Willebrand's factor (vWF, a marker of endothelial injury) or myeloperoxidase (a marker of neutrophil activation). DESIGN: Randomised experimental study. SETTING: University department of surgery, New Zealand. ANIMALS: Thirty anaesthetised adult Wistar rats. INTERVENTIONS: The sham operated group (n = 10) had laparotomy and isolation of the superior mesenteric artery without clamping. The ischaemia reperfusion group (IR, n = 10) had the superior mesenteric artery clamped for 1 hour and reperfusion for 15 minutes. The control group (n = 10) had direct puncture of the heart to sample blood. MAIN OUTCOME MEASURES: Plasma concentrations of protein carbonyl, vWF, and myeloperoxidase. RESULTS: Plasma protein carbonyl concentrations were significantly higher in the IR group than in the sham group (p < 0.02, Mann-Whitney test, median (range) 0.187 (0.141-0.242) compared with 0.144 (0.121-0.185) nmol/mg) and in the control group (p < 0.01, Mann-Whitney test, median (range) 0.187 (0.141-0.242) compared with 0.136 (0.108 0.175) nmol/mg). There was a significant correlation between protein carbonyl and vWF concentrations (r = 0.54, F = 10.9, p < 0.003, linear regression) but not with those of myeloperoxidase. CONCLUSION: Intestinal ischaemia-reperfusion caused an increase in the plasma protein carbonyl concentration, which is possibly produced by endothelial cells. PMID- 10365843 TI - Gracilis muscle flap in the treatment of persistent, infected pelvic necrosis. PMID- 10365844 TI - Familial hiatal hernia and gastro-oesophageal reflux disease. PMID- 10365845 TI - Videothoracoscopic excision of mediastinal parathyroid adenoma. PMID- 10365846 TI - Coronary atherosclerosis and interventions: pathological sequences and restenosis. AB - The primary cause of cardiac morbidity and mortality in developed countries is ischemic (coronary) heart disease. The incidence of this disease is virtually all due to atherosclerosis, and ischemic heart disease is also the most prevalent disease in the industrialized world, causing over 40% of all deaths in the United States and Western Europe. In Japan, the incidence of ischemic heart disease due to coronary atherosclerosis is gradually increasing as well. Compared with the classical nomenclature of atherosclerosis; that is, fatty streak, fibrous plaque and complicated lesions, the term Stary's classification has been universally accepted because it reflects the more recently acquired knowledge about the morphological and biochemical details of the processes in coronary atherosclerosis, which have been obtained by new strategies such as angioscopy, intravascular ultrasound and molecular biological methods. The term Stary's classification has been applied for the coronary atherosclerosis of patients with acute coronary syndrome at the National Cardiovascular Center, for the analysis of predisposing atherosclerosis of these patients. The recent findings regarding acute coronary syndrome resulting from a rupture of coronary atherosclerotic plaques indicate that this syndrome is probably the most important mechanism underlying the sudden onset. It has been found that the risk of plaque rupture may depend more on plaque composition than on plaque size. Plaques rich in soft extracellular lipids and macrophages are possibly more vulnerable to plaque rupture. Two of the goals of the present review are to clarify how plaque disruption occurs and to elucidate the relationship between plaque disruption and coronary risk factors in elderly Japanese patients with acute coronary syndrome. Coronary stents have been shown to be efficacious in the treatment of acute and threatened closure complicating percutaneous transluminal coronary angioplasty (PTCA) and have produced encouraging initial results in the prevention of restenosis. In the autopsy study of restenosis after PTCA, it was observed that dense caps of collagen fibers in the adventitia in the vicinity of the disrupted internal elastic laminae were present in all of the remodeling lesions. It is suggested that remodeling, which resulted in adventitial scarring, is one of the major causative factors of restenosis after PTCA. The long-term success of stenting, however, remains limited by the occurrence of late in-stent restenosis, with an incidence of 20-42% depending on the stent design and the patient population studied. Another aim of the present review is to describe the pathological mechanism of restenosis after PTCA and/or stent replacement and, consequently, the vascular remodeling that occurs around adventitial tissue after PTCA and intimal hyperplasia that is chronically irritated by a foreign body granulomatous reaction after stenting. Finally, the results of the investigation of the effect of a tissue factor pathway inhibitor on the prevention of interventional restenosis is described. PMID- 10365847 TI - Morphological characteristics of lipid accumulation in liver-constituting cells of acid lipase deficiency rats (Wolman's disease model rats). AB - Lysosomal acid lipase is a hydrolase essential for the intracellular degradation of cholesteryl esters and triglycerides. In the laboratory, rats with congenital deficiency of lysosomal acid lipase and marked accumulation of cholesteryl ester, cholesterol free and triglyceride in livers (Wolman's disease rat or Yoshida rat) that corresponded to human Wolman's disease were found and maintained. The morphological characteristics of accumulated lipids in the livers of affected rats were examined also. Many small lipid droplets and lipid crystals were found in the cytoplasms of hepatocytes and ED1-positive and ED2-positive foamy Kupffer's cells, respectively. Electron microscopically, many electron-lucent lipid droplets with limiting membrane were found in hepatocytes. Foamy Kupffer's cells had many multivesicular bodies with limiting membrane, which contained crivilinear bodies, lipid droplets and crystal clefts. At areas of aggregation of foamy Kupffer's cells forming islets, there were many desmin-positive Ito cells. Small lipid droplets with limiting membrane were also found in the cytoplasm of Ito cells and endothelial cells. These findings, which were obtained by morphological methods, indicated that triglyceride and both cholesteryl ester and free cholesterol accumulated in lipolysosomes mainly in hepatocytes and Kupffer's cells, respectively, and suggest that lysosomal acid lipase could participate in dissolution of the membrane. PMID- 10365848 TI - Synovitis in rheumatoid arthritis: scoring of characteristic histopathological features. AB - Synovial tissue specimens obtained from the knee joints of 40 patients with rheumatoid arthritis (RA) and from 22 patients with osteoarthritis (OA) were examined histologically. The histopathological features of RA synovitis and OA synovitis were then compared. Seven criteria items of histopathological features characteristic to RA synovitis were given a score of 1-3 points each in order to evaluate the histological severity of the seven items. Their total scores were then calculated. A comparison of the total RA synovitis score and the total OA synovitis score revealed that RA synovitis showed more than 11 points (maximum 20 points), while OA synovitis showed less than 10 points in all but two cases. Furthermore, the total scores of RA synovitis were then determined in the same manner for other joints, where it was confirmed that five other joints had scores of more than 11 points as well; that is, the intercarpal, wrist, elbow, ankle and hip joints in 52 patients with RA. From these results, it was concluded that in the histological examination of biopsied synovial tissue of RA, if the total score for synovitis is more than 11 points (maximum 20 points), an histological diagnosis of RA synovitis can be confirmed. PMID- 10365849 TI - Paracortical hyperplasia of superficial lymph nodes in a new mutant strain of hairless rats (ISh): a lesion similar to human dermatopathic lymphadenopathy. AB - The dermal histology and the regional draining superficial lymph node of a new mutant strain of hairless rats (ISh) were investigated. The homozygote ISh rat was characterized as having naked and wrinkled skin. The comedo-like cysts in the skin resembled human acne and vulgaris. Histopathologically, many Langerhans' cells positive for anti-protein kinase C type II antibody (PKC-II) were recognized in the epidermis. The superficial lymph nodes were significantly larger than those of the heterozygote. The paracortex of these lymph nodes was expanded and the number of Langerhans' cells mainly increased in these areas. These lymph node lesions were similar to those of human dermatopathic lymphadenopathy. The ISh rats should be a very useful animal model for studying dermatopathic lymphadenopathy in humans. Furthermore, they may be a valuable animal model for investigating the mechanisms of not only maturation, movement and migration of Langerhans' cells, but also of their function for antigen presentation. PMID- 10365850 TI - An immunohistochemical study of hepatic atypical adenomatous hyperplasia, hepatocellular carcinoma, and cholangiocarcinoma with alpha-fetoprotein, carcinoembryonic antigen, CA19-9, epithelial membrane antigen, and cytokeratins 18 and 19. AB - Eight hepatic atypical adenomatous hyperplasias (AH), 30 hepatocellular carcinomas (HCC) consisting of 11 well-, 13 moderately and six poorly differentiated HCC, and 10 intrahepatic cholangiocarcinomas (CC) were investigated immunohistochemically with anti-alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), CA19-9, epithelial membrane antigen (EMA), and cytokeratins (CK) 18 and 19 antibodies. Immunostaining was regarded as positive when more than 5% of cells were stained. Alpha-fetoprotein was positive, although focally, in five (17%) of 30 HCC but negative in all AH and CC. Carcinoembryonic antigen (polyclonal antibody) did not stain the cytoplasm of all AH and HCC, but stained two (25%) of eight AH and 10 (33%) of 30 HCC in a bile canalicular staining manner. Carcinoembryonic antigen showed intracytoplasmic or luminal border staining in six (60%) of 10 CC. CA19-9 was negative in all AH and HCC, while six (60%) of 10 CC were positive for CA19-9. Epithelial membrane antigen was positive in one (13%) of eight AH, seven (23%) of 30 HCC and in all 10 cases of CC. Cytokeratin 18 was positive in all AH, HCC and CC. Cytokeratin 19 was negative in both AH and HCC, whereas it stained the cytoplasm of tumor cells in all CC diffusely and intensely. These results suggest that immunostaining of AFP, CEA, CA19-9, EMA, CK18 and CK19 are not useful in the differential diagnosis between AH and well-differentiated HCC, and that CK19 is the most suitable reagent for the differential diagnosis between HCC and CC. PMID- 10365851 TI - Primary hepatic carcinoid and neuroendocrine carcinoma: clinicopathological and immunohistochemical study of five cases. AB - Primary hepatic carcinoid and neuroendocrine carcinoma (NEC) are rare tumors. We experienced three carcinoids and two NEC originating in the liver during the past 25 years and attempted to elucidate the clinicopathological and immunohistochemical features of these tumors. The patients had no endocrine symptoms despite two of them having elevated plasma serotonin. Three of the five patients died of the tumor after operation with an average survival time of 20.6 months. All tumors were large (up to 26 cm in diameter), four of them solitary and one multinodular, and were not associated with liver cirrhosis. The carcinoid tumors showed insular, trabecular or glandular arrangement of argyrophilic cells, whereas in the NEC this histological pattern was distorted. Immunohistochemically the tumors showed expression of chromogranin A (all cases), chromogranin B (three cases), pancreastatin and chromostatin (four cases, respectively), prohormone convertase PC3 (three cases), carcinoembryonic antigen (CEA) and CA19-9 (two cases), cytokeratin 56 kDa (three cases), 160 kDa neurofilament (two cases) and neuron-specific enolase (two cases). Serotonin and glucagon were sporadically detected in two tumors. The most useful marker to confirm the diagnosis was chromogranin A, which was cleaved to pancreastatin and chromostatin in the tumor tissue, and was more reliable than other markers of neuroendocrine differentiation. PMID- 10365852 TI - Expression of sialyl-Tn, Tn and T antigens in primary liver cancer. AB - Sialyl-Tn, Tn and T antigens are caused by aberrant or incomplete glycosylation of apomucins and are related to the aggressiveness of malignant neoplasms. Using 41 liver samples from patients with cholangiocarcinoma (including four with cirrhosis), 21 with combined hepatocellular-cholangiocellular carcinoma and 17 with hepatocellular carcinoma, the expression of sialyl-Tn, Tn and T antigens were characterized immunohistochemically and the correlation with apomucin profiles was evaluated. The prevalence of sialyl-Tn, Tn and T antigens expression was 89, 95 and 51% in cholangiocarcinoma without cirrhosis; 25, 75, and 0% in cholangiocarcinoma with cirrhosis; 29, 90, and 48% in combined hepatocellular cholangiocellular carcinoma; and 0, 12 and 6% in hepatocellular carcinoma, respectively. Sialyl-Tn antigen was frequently expressed in cholangiocarcinoma without cirrhosis compared with cholangiocarcinoma with cirrhosis and combined hepatocellular-cholangiocellular carcinoma (P < 0.01). Although sialyl-Tn expression was associated with MUC1, MUC6 and MUC7 expression, the expression sites among them were not identical in the individual cases. These data suggest that the different expressions of sialyl-Tn antigen among cholangiocarcinoma without cirrhosis, cholangiocarcinoma with cirrhosis and combined hepatocellular cholangiocellular carcinoma may reflect the biological features inherent to these tumors, such as the ability of invasion. PMID- 10365853 TI - An evaluative system for the response of antibacterial therapy: based on the morphological change of Helicobacter pylori and mucosal inflammation. AB - The diagnostic standard is an important factor in the evaluation of the antibacterial effect to Helicobacter pylori (H. pylori). Few studies have evaluated the bacterial morphological change. In the present study, H. pylori was examined by means of electron microscopy (EM), light microscopy (LM) and immunohistochemical staining. Patients were followed up from 6 weeks to more than 1 year after treatment for H. pylori, and the results of the 13C-urea breath test (UBT) were compared. A '4-L' evaluative system was used for histological diagnosis; that is, complete, significant, partial and negative response for H. pylori treatment. Complete response showed no H. pylori in histology, and positive 13C-UBT and negative response showed positive in both diagnoses. A significant response showed the morphology of H. pylori was thick walled by EM, that there was no obvious active inflammation, and was negative for C-UBT. These H. pylori showed a coccoid form and possibly static bacteria, which was resistant to further antibacterial therapy. The '4-L' system could evaluate the antibacterial effect, suggesting the necessity for a second line of therapy for H. pylori. It is suggested that this sensitive evaluative system is suitable for clinical applications for antibacterial therapy. PMID- 10365854 TI - A retroperitoneal bronchogenic cyst with malignant change. AB - A unique case of adenocarcinoma arising in a retroperitoneal bronchogenic cyst is presented. A 55-year-old woman presented with lower abdominal discomfort. Computed tomography revealed a retroperitoneal cystic mass attached to the ascending colon. The resected cyst was unilocular and filled with milky white mucus and hemorrhagic debris. Histologically, most of the cyst wall was of well differentiated papillary adenocarcinoma with no cyst wall invasion. Other small areas of the cyst were lined with variably atypical dysplastic/metaplastic cuboidal to pseudostratified columnar epithelium. The cyst wall was mostly hyalinized, but there was apparent thickened subepithelial basement membrane, elastosis, and a single layer of smooth muscle that suggested bronchial wall structures. A mucin staining study with O-acylated sialic acid, which is used for the demonstration of gastrointestinal, cholecystic and uterine cervical mucins, was negative for the mucin-producing epithelial cells of the cyst. Thus, to our knowledge, this is the first reported case of adenocarcinoma arising in a retroperitoneal bronchogenic cyst. PMID- 10365855 TI - Primary giant cell malignant fibrous histocytoma of the lung: a case report. AB - A rare case of malignant fibrous histiocytoma of giant cell type originating in the lung of a 46-year-old woman is presented. The patient complained of having a cough that had lasted for a few weeks. A chest X-ray photograph showed a tumor shadow on the left lung. Histological and cytological examination of the biopsy specimen revealed that the tumor was a kind of sarcoma. An operative procedure was selected because of tumor invasion into the trunk of the left pulmonary artery, which was discovered on computed tomography examination, and because metastatic tumor was excluded clinically. The tumor was almost encapsulated and 6 x 6 x 6 cm in size; however, it also showed invasion into the pulmonary artery and bronchial lumen. A histological survey of the tumor showed a wide range of patterns such as fibrous, pleomorphic, fascicular and osteoclast-like giant cell figures; however, the osteoclast-like giant cell area was predominant. Immunohistochemically, the tumor cells were positive for vimentin, CD68 for histiocytic marker and alpha1-antichymotrypsin, and negative for keratin, epithelial membrane antigen, S-100 protein, MT-1, desmin, myoglobin and lysosome. No primary tumor was found clinically in any part of the patient's body at 2 and 4 months after operation. Consequently, she was diagnosed as having primary giant cell malignant fibrous histiocytoma of the lung. PMID- 10365856 TI - Cutaneous granulocytic sarcoma mimicking immunoblastic large cell lymphoma. AB - A peculiar case of cutaneous granulocytic sarcoma without leukemic manifestation (so-called aleukemic leukemia cutis) that developed in the skin of the back of a 69-year-old man is reported. A skin biopsy specimen showed atypical cells with a prominent nucleolus proliferating around dermal blood vessels and along adnexa without epidermotropism. Atypical cells similar to those of the skin had infiltrated diffusely into the interfollicular area of an inguinal lymph node. Flow cytometric and immunohistochemical studies with a panel of monoclonal antibodies revealed neoplastic cells that had a biphasic phenotype of myeloid and T cell precursors. They expressed CD13, CD15, CD33, lysozyme, CD3epsilon, CD4, CD7 and terminal deoxynucleotidyl transferase (TdT). Gene analysis showed no rearrangement of the immunoglobulin heavy chain or T cell receptor beta and gamma genes. Ultrastructurally, the tumor cells exhibited a few intracytoplasmic electron-dense granules and well-developed rough endoplasmic reticulum with an occasional whorling arrangement. The initial diagnosis was immunoblastic large cell lymphoma, and the patient was treated with six courses of ProMACE-CytaBOM. In spite of the high-grade cytological characteristics of this tumor, the patient has been free of disease for 5 years. PMID- 10365857 TI - Cutaneous ciliated cyst of the right lower leg. AB - A 23-year-old Japanese woman with a cutaneous ciliated cyst on her right lower leg is reported. A subcutaneous cyst, measuring 2.5 cm in diameter with papillary projections into the lumen, was lined with ciliated cuboidal to columnar epithelia with partial stratification, histologically. These lining cells did not produce mucin. Immunohistochemically, the ciliated lining cells of the cyst were diffusely positive to epithelial membrane antigen and cytokeratin. In addition, positive immunoreaction with anti-desmin monoclonal antibody was observed in the body of the cilia. Less than 10% of the epithelial cells revealed positive immunoreaction to S-100 protein and estrogen receptor. PMID- 10365858 TI - Esophageal undifferentiated carcinoma displaying marked chondroid differentiation at metastatic foci. AB - A report of an unusual esophageal tumor in an 81-year-old man is presented. The primary tumor was diagnosed as undifferentiated carcinoma at biopsy and had disappeared after irradiation treatment. However, multiple metastases were noted in the brain, lungs, kidneys, adrenals and spleen at autopsy. Histologically, metastases showed marked cartilaginous metaplasia as demonstrated by light microscopy, histochemical and immunohistochemical studies, although the initial biopsy sample did not possess chondroid matrix. Furthermore, an apparent transition could be traced from carcinomatous to chondroid cells, suggesting that the chondroid cells were derived from carcinoma cells. The carcinomatous area partially showed both squamous and glandular differentiation, although they were poorly differentiated. A retrospective immunohistochemical study that used a panel of antibodies suggested a phenotypic relevance between primary and metastatic tumors. PMID- 10365859 TI - Heterotopic sebaceous glands in the esophagus: histopathological and immunohistochemical study of a resected esophagus. AB - A resected esophagus with numerous heterotopic sebaceous glands was examined in an attempt to determine whether esophageal heterotopic sebaceous glands are the result of a metaplastic process or a congenital anomaly. The present case concerns a 79-year-old Japanese man with numerous esophageal heterotopic sebaceous glands accompanied by superficial esophageal cancer. The resected esophagus possessed numerous heterotopic sebaceous glands, which could be seen clearly as slightly elevated, yellowish lesions. Histological examination of these glands, all of which were located in the lamina propria, revealed lobules of cells that showed characteristic sebaceous differentiation. Bulbous nests of proliferating basal cells showing sebaceous differentiation were occasionally observed in the esophageal epithelium. Of the antibodies against six different keratins used, only anti-keratin 14 labeled both the heterotopic sebaceous glands and the bulbous nests. Acquired metaplastic change of the esophageal epithelium is probably the pathogenetic mechanism involved in these unusual lesions. PMID- 10365860 TI - Evidence-based clinical practice, [corrected] evidence-based medicine and the Cochrane collaboration. AB - Encouraging professionals in training and later to consider practice-related research findings when making important clinical decisions is an on-going concern. Evidenced-Based Medicine (EBM) and the Cochrane Collaboration (CC) provide a source of tools and ideas for doing so, as well as a roster of colleagues who share this interest. Evidenced-based medicine involves integrating clinical expertise with the best available external evidence from systematic research as well as considering the values and expectations of patients/clients. Advantage can be taken of educational formats developed in EBM, such as problem based learning and critical-appraisal workshops in which participants learn how to ask key answerable questions related to important clinical practice questions (e.g., regarding effectiveness, accuracy of assessment measures, prediction, prevention, and quality of clinical practice guidelines) and to access and critically appraise related research. The Cochrane Collaboration is a world-wide network of centers that prepare, maintain, and disseminate high-quality systematic reviews on the efficacy of healthcare. These databases allow access to evidence related to clinical practice decisions. Forging reciprocal working relationships with those involved in EBM reciprocal and the CC should contribute to the pursuit of shared goals such as basing clinical decisions on the best available evidence and involving clients as informed consumers. PMID- 10365861 TI - Using a single-subject design to assess the development of anxiety in humans. AB - Laboratory studies of conditioned anxiety using a between-subjects group design typically involve only one session of conditioning trials. Because research suggests that anxiety disorders develop and change with repeated exposure to aversive events, these studies may fail to mimic the developmental course of anxiety in the natural environment. We highlight the limitations of single session studies for examining the development of conditioned anxiety within individual participants and discuss how many of these limitations can be avoided using single-subject designs. Furthermore, we suggest that the most comprehensive account of anxiety can be attained by using a single-subject design in conjunction with more commonly used group designs. PMID- 10365862 TI - Commentary: On the limited role of the "single-subject" design in psychology: hypothesis generating but not testing. AB - The "single-subject" design (which really denotes a design that employs too few subjects to allow statistical inferences concerning significance to be made) is useful only for the generation, but not for the testing or evaluation, of hypotheses concerning any psychological function. Those hypotheses that may be suggested by the "single-subject" design include ones about within-subject developmental effects (which may need to be studied over multiple sessions), as well as individual-difference-related variables. To adequately test any of those hypotheses, however, it is necessary to employ designs that vary both within- and between-subject factors, and that also examine various correlations in such a way that all effects (including correlational ones) can be evaluated by conventional modes of statistical inference. PMID- 10365863 TI - The relationship between anxious rearing behaviours and anxiety disorders symptomatology in normal children. AB - The present study examined the relationship between perceived parental rearing practices, in particular anxious rearing behaviours, and anxiety symptoms. One hundred and seventeen school children aged between 9 and 12 years completed the EMBU for children, a questionnaire that measures perceptions of parental rearing behaviours, and the Children's Anxiety Scale, an index of DSM-defined anxiety disorders symptoms. Results showed that there were significant and positive associations between Parental Rejection, Anxious Rearing, and Parental Control, on the one hand, and anxiety symptoms, on the other hand Emotional Warmth was not related to anxiety symptoms. PMID- 10365864 TI - A comparison of people with and without nocturnal panic attacks. AB - This study examined differences in frequency and severity of diurnal panic attacks between patients with (n = 22) and without (n = 21) a history of nocturnal panic attacks. Subjects were assessed with a modified version of the SCID, and completed daily panic attack diaries, the Anxiety Sensitivity Index, and the Fear Questionnaire. No differences were found between the groups in the actual number of expected or unexpected diurnal panic attacks experienced. Subjects in the nocturnal panic group experienced significantly more symptoms during diurnal panic attacks than did the diurnal only group. More specifically, the nocturnal panic group experienced more symptoms during expected diurnal panic attacks, but not during unexpected/spontaneous diurnal panic attacks. A greater proportion of the nocturnal panic subjects reported "chest pain" during diurnal panic attacks and a trend toward greater "fear of dying". Otherwise, the two groups were very similar. Implications of the findings are discussed in relation to the findings of other recent studies of nocturnal panic. PMID- 10365865 TI - Animal models of psychopathology: the establishment, maintenance, attenuation, and persistence of head-banging by pigeons. AB - Investigators of animal models of psychopathology have typically introduced experimental conditions so that an animal's behavior progressively deviates from a baseline of routine laboratory behavior toward a pattern which resembles human psychopathological behavior in some form of S, then R relation. The present experiments report consequential contingency procedures for bringing head-to-wall head-banging by an animal under experimental control and analysis. The first two experiments examined the establishment and maintenance by reinforcement of head banging by pigeons. The final two experiments examined the occurrence of head banging, under conditions of extinction and limited reinforcement, when an alternative behavior, i.e., key-pecking, was reinforced under a variety of reinforcement schedules. Extinguished and infrequently reinforced head-banging was found to recur under a variety of conditions including the reinforcement of the more "normal" alternative behavior. To the extent that human patterns are governed by similar functional relations, the data may be of relevance in the analysis of the maintenance, attenuation, and recurrence of human patterns designated as pathological. Further, the permanent elimination of a disturbing pattern may be difficult, and the recurrence of a disturbing pattern might properly be considered a likely and "normal" outcome of basic behavioral processes. PMID- 10365866 TI - Using item analysis to facilitate interpretation of empirical findings. AB - Researchers often present and interpret empirical findings with reference to hypothetical constructs and diagnostic labels. Such interpretations commonly are based upon "summary" scores obtained through interview, self-report, or rating scale assessment instruments. Although there are advantages associated with communicating empirical findings through analysis with summary scores, there also are weaknesses that may limit the interpretability of empirical findings and impede theory development. We discuss the importance of item analysis as a tool that may guide presentation of empirical findings, and we describe how it may be used to minimize these limitations of assessment, facilitate data interpretation, and increase the opportunity for theoretical advances. PMID- 10365867 TI - Presidential address. Surgeons and technology. PMID- 10365868 TI - Repair of paraesophageal hernias. AB - BACKGROUND: Three years ago we proposed the use of laparoscopy and systematic addition of an antireflux procedure to repair paraesophageal hernias. We now present an analysis of the outcome on patients and the evolution of the technique proposed. METHODS: Symptoms and esophageal function were prospectively collected and followed in 41 consecutive patients treated over a 4-year period. Indications for repair included chronic anemia in 15 patients, and previous incarceration in 8. Twenty-two patients had symptoms of reflux. RESULTS: All operations were started laparoscopically, two were converted. Mean operating time was 210 minutes, and mean hospital stay was 4 days. Mean follow-up was 3 years. The operation was effective; all symptoms had improved significantly at last follow up. CONCLUSIONS: Laparoscopic repair of paraesophageal hernia with the addition of an antireflux procedure, although difficult, lengthy, and not totally without risk, improves symptoms substantially, resolves anemia, and prevents incarceration in nearly all patients. PMID- 10365869 TI - Esophageal motility and outcomes following laparoscopic paraesophageal hernia repair and fundoplication. AB - BACKGROUND: The addition of an antireflux procedure to all giant paraesophageal hernia (PEH) repairs remains controversial. In addition there are no series evaluating the impact of hernia repair and fundoplication on esophageal physiology. This study examines the outcomes of PEH repair with fundoplication and examines the results of preoperative and postoperative motility and pH testing. METHODS: An analysis of a data base containing all patients undergoing PEH repair between September 1994 and December 1997. Patients underwent laparoscopic sac reduction, hernia repair, and fundoplication. Follow-up was performed under protocol and consisted of a symptoms assessment form, 24 hour pH, and manometry. RESULTS: Fifty-two patients (mean age 63) were treated: 59% complained of heartburn, 50% dysphagia, and 27% chest pain; 26% had a body motility disorder. Complete manometry was not possible in 41%. Mean operative time was 4 hours. There were 48 Nissen, 4 Toupet, and 7 Collis-Nissen procedures. There were 3 (6%) intraoperative and 3 (6%) postoperative complications. There were no operative mortalities. Hospital stay was 3 days (1 to 29). Late follow-up (18 months) was available for 96% of patients and showed dysphagia in 6%, heartburn in 10%, and recurrent herniation in 8%. Objective postoperative testing was available in 61 % of the patients at a mean of 8 months. Twenty-four hour pH tests were abnormal in 4 patients (2 asymptomatic and 2 with a Collis). Lower esophageal sphincter pressures increased 63% and functioned well in 71% of patients; 50% of preoperative motility disorders improved following repair. CONCLUSIONS: Laparoscopic repair of giant PEH is technically difficult but feasible. Routine addition of a fundoplication is advised, as preoperative testing is unreliable for a selective approach and fundoplications are well tolerated in this group of patients. PMID- 10365870 TI - Cholecystectomy is becoming an increasingly common operation in children. AB - PURPOSE: To determine the cause of a marked rise in cholecystectomy at a regional children's hospital. METHODS: Retrospective review of 185 patients undergoing cholecystectomy since 1984. The years 1984 to 1990 (group I) and 1991 to 1996 (group II) were compared. RESULTS: Cholecystectomy for gallbladder disease increased from 4.4/year (group I) to 16.3/ year (group II). Abdominal ultrasound examinations increased during this time. The ratio of children diagnosed with gallstones and then undergoing cholecystectomy also increased (P = 0.005). In group 11, 43% of children had no apparent etiology for gallstones, and more children developed complications of gallstones and evidence of choledocholithiasis. CONCLUSIONS: (1) The increased incidence of cholecystectomy is probably multifactorial. (2) Gallstone identification has increased owing to increased patient visits and more liberal use of ultrasonography in patients with abdominal pain. (3) More patients with cholelithiasis now undergo cholecystectomy perhaps because of a change in physician perception of the disease and an apparent increase in complications from gallstones. PMID- 10365871 TI - Is laparoscopic donor nephrectomy here to stay? AB - BACKGROUND: Open live donor nephrectomy is safe and provides kidneys of excellent quality. The complexity of the laparoscopic donor technique has raised considerable concerns. METHOD: Twenty-six laparoscopic live donor nephrectomies were done from October 1997 to October 1998. RESULTS: All kidneys had immediate function. All recipients except 1 had serum creatinines less than 2.0 mg at 2 months posttransplantation. Three complications (wound infection, neuroma, reoperation) occurred. There was no mortality. CONCLUSIONS: Proper surgical training and patient selection can result in a safe donor operation that provides kidneys of excellent quality. PMID- 10365872 TI - Urban trauma care is threatened by inadequate reimbursement. AB - BACKGROUND: Hospitals struggle to support trauma care. Recent installation of cost accounting systems now provides information on actual costs for different categories of patients. This paper examines the cost of trauma care in an urban teaching hospital. METHODS: All patients entered into the hospital trauma registry for the period July 1, 1996, through June 30, 1997, were abstracted from the registry. These data were merged with a database of all admitted patients with an injury-compatible ICD-9 diagnostic code for the same time period that included cost and estimated revenue from the cost accounting system. Complete data were available for 667 patients and the remaining 96 were uninsured patients with missing cost data. RESULTS: The calculated cost of care for the 667 patients was $10,342,130; total expected revenue was $10,396,456; estimated net revenue for insured patients was $54,326. The estimated cost of care for the 96 uncompensated patients was $1,619,989. The hospital had positive net revenue for patients with length of stay of 7 days or less, but was unable to recoup costs for patients with a longer stay. Reimbursement exceeded hospital cost for blunt injuries, primarily motor vehicle crash victims, and for other injuries covered by fee-for-service insurers. Managed care plans and government-funded insurance did not reimburse sufficiently to cover hospital costs. CONCLUSIONS: These data confirm that earlier literature, based on charges and estimated costs, were correct in documenting a serious threat to the continuation of centers providing high volumes of trauma care. PMID- 10365873 TI - The role of computed tomography with contrast and small bowel follow-through in management of small bowel obstruction. AB - BACKGROUND: In a significant percentage of patients, radiologic evaluation other than plain abdominal films are required to confirm or exclude the presence of small bowel obstruction. METHODS: Over a 1-year period, 55 patients had both computed tomography and small bowel follow-through studies. Patients were classified as having (1) paralytic ileus, (2) low-grade obstruction, (3) high grade obstruction, or (4) complete mechanical obstruction. The gold standard for diagnosis was celiotomy in 42 patients and clinical follow-up in 13 patients. RESULTS: Thirty-six out of 42 patients had proven intestinal obstruction at the time of celiotomy. Computed tomography identified 32 out of the 36 high-grade and complete mechanical obstructions. Computed tomography was superior to small bowel follow-through in identifying masses, malignancies, and features of strangulation. Small bowel follow-through correctly identified "insignificant obstructions" when contrast reached the cecum within 4 hours in 18 of 19 patients. CONCLUSIONS: In patients with equivocal findings of small bowel obstruction, computed tomography should be used initially and then small bowel follow-through if computed tomography is not diagnostic. Computed tomography was superior in this study for detecting the cause of the intestinal obstruction and presence of strangulation. PMID- 10365874 TI - Stages at presentation, prognostic factors, and outcome of breast cancer in males. AB - BACKGROUND: In order to support or refute conventional notions of breast cancer in males as a late-presenting disease associated with a worse prognosis than the same disease in females, we reviewed a recent, multi-institutional experience. METHODS: A case series from three area hospital system cancer data bases was reviewed. Demographics, pathology, stages at presentation, and treatment were determined from the data set and correlated with outcomes (recurrence/survival). RESULTS: Fifty-four patients (mean age 64.5, SD = 12.8) were identified; half of the tumors were stage T0 or T1, 62% were node negative (N0), and 57% had an American Joint Committee on Cancer (AJCC) stage grouping of IIA or less. Eighty five percent of tumors examined expressed hormone receptors. There were no local only recurrences in the 50 cases resected for cure, including 5 cases of minimal breast cancer treated by lumpectomy only. Five- and 10-year overall disease-free survival was AJCC stage related: 100% and 71%, respectively, for early stage (0 IIA) disease, and 71% and 20%, respectively, for advanced (IIB-IV) stage (P = 0.0051 by log-rank). Only AJCC stage and its components (tumor size, nodal status, presence of metastases) correlated with survival by multivariate analysis; other factors such as age, family history, and presenting symptoms/signs did not. CONCLUSIONS: The majority of breast cancers in males present at early stages and are hormone receptor positive. In contrast to older notions of this disease as uniformly aggressive, we conclude that prognostic factors and stage-for-stage outcomes for breast cancer in males are similar to those published for the disease in females. PMID- 10365875 TI - Pathologic risk factors of occult malignancy in endoscopically unresectable colonic adenomas. AB - BACKGROUND: With the advent of new endoscopic and laparoscopic techniques, the likelihood of colonoscopically unresectable adenomas harboring occult malignancy influences management. While prior studies have evaluated polyp size and morphology in assessing the risk of malignancy, the relative risk of cancer based on the presence or absence of high-grade dysplasia has not been systematically studied. METHODS: For all lesions preoperatively diagnosed as adenomas without invasive cancer, multivariate logistic regression analysis was performed to elicit independent variables associated with malignancy in the resected specimen. RESULTS: One hundred patients underwent a colectomy for preoperatively diagnosed adenomatous lesions. Multivariate logistic regression analysis revealed size, degree of dysplasia, and left-sided location to be independent predictors of malignancy. CONCLUSIONS: In colonic adenomas which are not amenable to simple colonoscopic resection, the most useful predictors of the lesion harboring a malignancy are polyp size and the presence of high-grade dysplasia, and these factors can help determine management. PMID- 10365877 TI - Recurrence and survival after surgical management of rectal cancer. AB - BACKGROUND: Reported local recurrence rates for rectal cancer are significantly reduced using a combination of superior surgical technique, in the form of total mesorectal excision, and routine radiotherapy. In an attempt to determine the effectiveness of current local management strategies, a review of Vancouver Island Cancer Centre patients with rectal cancer was performed and the overall local recurrence rate was identified. METHODS: We retrospectively reviewed the charts of 272 rectal cancer patients from 1988 to 1998. Two hundred and twenty nine patients met inclusion criteria. Analysis of patient factors included age, gender, type of surgery, and adjuvant therapy. Tumors were assessed for level, stage, and grade. Local recurrence and distant metastases were also documented. Variables influencing local recurrence in this group were identified and disease free and actuarial survival determined. RESULTS: Of 229 patients analyzed, 12.7% (29) had local recurrences. Variables influencing local recurrence were number of positive lymph nodes, vascular invasion, and neural invasion. There was no significant difference in local recurrence between patients having anterior resection and those having abdominoperineal resection. None of the patients who received preoperative radiotherapy had a local recurrence. Actuarial disease-free survival was 87% at 5 years. CONCLUSIONS: Limiting local recurrence is one of the most important goals in the treatment of rectal cancer. It is essential to identify those patients with "high risk" tumors as identified by endorectal ultrasound or pathologic features. These patients comprise the group most likely to benefit from a routine mesorectal excision combined with adjuvant radiotherapy. PMID- 10365876 TI - Effects of endorectal ultrasonography in the surgical management of rectal adenomas and carcinomas. AB - BACKGROUND: Accurate preoperative staging of rectal lesions can help select patients for treatment options. This prospective study examines the effects of endorectal ultrasonography (ERUS) in the management of resectable adenomas and adenocarcinomas. METHODS: Between November 1995 and August 1998, 34 patients had resection of their rectal lesions (28 adenocarcinomas, 6 adenomas). Preoperative clinical stage and surgical and adjuvant therapy plans were specified before ERUS staging; then comparisons were made to postoperative pathologic stage and actual treatment. RESULTS: ERUS helped select 7 of 11 patients (64%) for preoperative chemoradiation therapy. Treatments of adenocarcinomas were altered by ERUS to local excision (LOC) in 4 of 9 patients (45%) planned for low anterior resection (LAR), and 2 of 10 patients (20%) planned for abdominoperineal resection (APR). ERUS helped spare 2 patients with adenoma from APR. CONCLUSION: ERUS is a useful modality in the surgical management of patients with rectal adenomas and carcinomas. PMID- 10365878 TI - Prospective evaluation of formalin therapy for radiation proctitis. AB - BACKGROUND: Radiation proctitis is a troublesome complication of radiation therapy for as many as 75% of patients after pelvic irradiation. Five percent progress to chronic radiation proctitis complicated by telangiectasias and hemorrhage. The utility of formalin rectal instillation for treatment of bleeding is prospectively evaluated in this study. METHODS: Eleven patients (9 male, 2 female) with rectal bleeding after pelvic irradiation were treated with formalin therapy. In a single treatment, 4% formalin was instilled into the rectum in four separate 20-cc aliquots with total mucosal contact time of approximately 15 minutes. Patients were initially evaluated at 7 to 10 days and 1 month postoperatively and assessed for bleeding. RESULTS: All patients presented with rectal bleeding. Twenty-seven percent required transfusion. Thirty-six percent had failed other previous therapy. In follow-up of 3 to 64 months, 100% had initial success with cessation of bleeding. Three patients had recurrent bleeding; none required transfusion. One patient required repeat formalin instillation, with no further bleeding at 3 months follow-up. CONCLUSION: Local rectal instillation of 4% formalin is an efficacious therapy for treatment of radiation-induced lower gastrointestinal bleeding. PMID- 10365879 TI - A prospective, randomized, double-blinded, placebo-controlled trial of cisapride after colorectal surgery. AB - BACKGROUND: The predominant factor prolonging hospitalization and delaying oral intake after colorectal surgery continues to be return of large bowel function. We investigated the effect of the cisapride on postoperative bowel motility. METHODS: Patients were started on cisapride versus a placebo on postoperative day 1 after colorectal surgery. Endpoints included the time to patients' first bowel movement, time of advancement to regular diet, time of hospitalization, and cost analysis. Results were analyzed using a Mann-Whitney test. RESULTS: Thirty-five patients were entered in the study, consisting of 17 in the cisapride group and 18 in the placebo group. The median time to first bowel movement, advancement of diet, and hospital discharge was 1 day less in the cisapride group compared with the placebo group (P <0.05). There was a substantial cost savings in the study group. CONCLUSIONS: Cisapride use results in statistically significant improvement in postoperative bowel motility. Cisapride should be added as adjunct treatment in postoperative care after colorectal surgery. PMID- 10365880 TI - Benefits and safety of hepatic resection for colorectal metastases. AB - BACKGROUND: Metastatic colorectal carcinoma to the liver is a potentially curable disease. The purpose of this study was to determine the safety and efficacy of hepatic resection for metastatic colorectal carcinoma. METHODS: One hundred twenty-one consecutive hepatic resections in 110 patients with metastatic colorectal cancer between January 1978 and September 1998 performed by a single surgeon were reviewed. RESULTS: The actuarial 5-year survival for all patients in the series was 46%. Of the patients operated on before 1993, the actual 5-year survival was 43% and actual disease-free 5-year survival was 28%. The actual 10 year survival was 27%, and of all patients operated on in the last 20 years, 48% are alive today. When comparing initial regional lymph node status, the 5-year survival was 54% for the patients with negative lymph nodes and 40% for patients with positive nodes. Only 18% of patients required a perioperative blood transfusion, and the median length of stay was 7 days. There were complications in 34% of cases, and the operative mortality was 4%. CONCLUSIONS: Hepatic resection for metastatic colon cancer is safe, and significant longevity and cure can be obtained after resection. PMID- 10365881 TI - Transcatheter arterial chemoembolization as primary treatment for hepatocellular carcinoma. AB - BACKGROUND: Hepatocellular carcinoma (HCC) in Western populations has historically been associated with poor survival. METHODS: In this study, we conducted a 7-year retrospective analysis of patients with HCC undergoing transcatheter arterial chemoembolization (TACE) at our institution and examined demographics, outcomes, and complications. RESULTS: During the period of study, 39 patients (25 male [64%], mean age 58 [range 17 to 86]) underwent a total of 78 chemoembolization treatments. During the same time period, an additional 31 patients received supportive care only. The majority of patients had late stage disease (American Joint Committee on Cancer stage III, IVa, or IVb) with no statistical difference noted between the two groups (P = 0.2). However, patients receiving supportive care only had significantly worse hepatic dysfunction by Child's classification (P = 0.005). Twenty-nine patients (74%) had documented cirrhosis, with hepatitis C being the most common cause in 11 of 29 (38%). In patients undergoing TACE, overall actuarial survival was 35%, 20%, and 11% at 1, 2, and 3 years with a median survival of 9.2 months, significantly improved over the group receiving supportive care only (P < 0.0001). Median survival for the group receiving supportive care was less than 3 months. Neither age nor stage had a significant impact on survival. The most common complications of TACE included transient nausea, abdominal pain, vomiting, and fever. CONCLUSIONS: TACE is a safe and effective therapeutic option for selected patients with HCC not amenable to surgical intervention. PMID- 10365882 TI - Radiofrequency ablation followed by resection of malignant liver tumors. AB - BACKGROUND: Radiofrequency ablation (RFA) has recently been used to treat liver tumors, but few clinical reports have described the pathological characteristics of radiofrequency ablation in human specimens. This study delineates the gross pathologic and histochemical changes induced by RFA in benign and malignant human liver tissue and confirms the tumor necrosis described in early clinical reports. METHODS: Ten patients with metastatic tumors of the liver received a single treatment of ultrasound-guided percutaneous RFA to 12 tumors. Hepatic resection was carried out within 6 weeks of RFA. Specimens were stained with standard hematoxylin and eosin stain followed by oxidative stain to determine if there was evidence of viable tumor within the zone of ablation. RESULTS: Nine of the 12 ablations were resected. Microscopic examination within the zone of ablation showed successful ablation in 8 of the 9 resected ablations. CONCLUSIONS: Percutaneous RFA creates well-circumscribed areas of tumor necrosis with apparent cell death using an oxidative stain. Further investigation is encouraged to determine the clinical effectiveness of radiofrequency ablation in the complete destruction of liver tumors for palliative or curative intent. PMID- 10365883 TI - Vancomycin-resistant Enterococcus in liver transplant patients. AB - BACKGROUND: Vancomycin-resistant Enterococcus (VRE) infection is emerging in the transplant population, and there is no effective antibiotic therapy available. The aims of this retrospective review were to (1) investigate the outcome of and (2) identify common characteristics associated with VRE infection and colonization in orthotopic liver transplant (OLTx) candidates. METHODS: From October 1994 through September 1998, 126 isolates of VRE were identified in 42 of 234 OLTx recipients and 5 OLTx candidates who did not proceed to transplantation. Data were collected by patient chart review or from a computerized hospital database. RESULTS: The 1-year mortality rate with VRE infection was 82%, and with VRE colonization, 7%. This mortality rate contrasts with a 14% 1-year mortality for non-VRE transplant patients (P <0.01, infected patients and colonized patients). Characteristics of VRE colonized and infected patients included recent prior vancomycin (87%), coinfection by other microbial pathogens (74%), recent prior susceptible enterococcal infection (72%), concurrent fungal infection (62%), additional post-OLTx laparotomies (47%), and renal failure (Cr >2.5 mg/dL or need for dialysis; 43%). Biliary complications were seen in 52% of post-OLTx VRE-infected or VRE-colonized patients (versus 22% in non-VRE transplant patients, P <0.05). CONCLUSION: VRE infection is associated with a very high mortality rate after liver transplantation. The incidence of biliary complications prior to VRE isolation is very high in VRE-infected and VRE colonized patients. The most common characteristics of VRE patients were recent prior vancomycin use, recent prior susceptible enterococcal infection, coinfection with other microbial pathogens, and concurrent fungal infection. With no proven effective antimicrobial therapy for VRE, stringent infection control measures, including strict and limited use of vancomycin, must be practiced. PMID- 10365884 TI - Distant processing of pancreas islets for autotransplantation following total pancreatectomy. AB - BACKGROUND: Small duct chronic pancreatitis is associated with intractable pain and failure to thrive, usually unresponsive to conventional management approaches. Total pancreatectomy is considered after failure of medical intervention. The major morbidity following total pancreatectomy is diabetes mellitus with its associated complications. This adverse outcome can be mitigated through autotransplantation of islets recovered from the pancreatectomy specimen. This approach has been limited historically owing to the absence of an on-site islet processing facility. We present the results from 5 pancreatectomized patients whose islets were prepared 1,500 miles away. METHODS: Five patients (4 women, 1 man, average age 42 years) who failed medical therapy and were not candidates for longitudinal pancreaticojejunostomy underwent total/completion pancreatectomy (4 total, 1 completion) for intractable symptoms from idiopathic small duct chronic pancreatitis. The resected pancreata were preserved in ViaSpan solution and were transferred to an islet processing laboratory by commercial airliner and returned. The dispersed pancreatic islet tissue was infused into a portal vein tributary through an operatively placed catheter after systemic heparinization. RESULTS: All 5 patients experienced complete relief from pancreatic pain; 2 had significant residual discomfort from underlying Crohn's disease. Three of the 5 patients had minimal or no insulin requirement after autotransplantation (median follow-up of 23 months); 1 patient continued with glycemic control difficulties related to Crohn's disease. One patient died 17 months following autotransplantation from an unrelated pneumonia. CONCLUSION: Total pancreatectomy with autologous islet transplantation can offer patients with idiopathic small duct chronic pancreatitis pain relief without the sequelae of diabetes mellitus and can be performed without an on-site islet processing facility. All patients undergoing total/ completion pancreatectomy should be considered candidates for this procedure. PMID- 10365885 TI - How accurate is helical computed tomography for clinical staging of pancreatic cancer? AB - BACKGROUND: Our goal was to determine if findings on an index computed tomography (CT) scan would correlate with survival in patients with pancreatic adenocarcinoma. We know that as this tumor extends out of the gland, survival decreases. Are there any CT findings that assess tumor extension sufficiently that also correlate with survival? Once identified, these CT areas would be the best factors to clinically stage patients. METHODS: Between 1993 and 1997, 160 patients with biopsy-proven adenocarcinoma of the pancreatic head were included if an index helical CT scan and clinical follow-up were available. All CT scans were reviewed by the same radiologist blinded for outcomes. CT scans were interpreted using a graded extension of tumor out of the pancreatic head in four areas: retroperitoneum (RP); anterior pancreatic capsule (S); portal/superior mesenteric veins (PV); and celiac/superior mesenteric arteries (A). Extension of tumor was graded as follows: Grade 0 (negative margin); 1 (suspicious); 2 (positive); or 3 (extensively involved). Also recorded and graded were signs of metastases: nodal enlargement > or =1.5 cm (N); and lesions consistent with hepatic metastases (H). Survival was compared between grades for each CT area using the methods of Kaplan and Meier and relative risk estimates of death (Cox regression models). RESULTS: Compared with grade 0, the following CT areas had significantly decreased survival curves: grade 1 (only S and A), grade 2 and 3 (RP, PV, S, A). N and H did not correlate with survival unless > or =1.5 cm nodes were in the liver or splenic hilum or there were multiple liver nodules. CONCLUSION: Although postoperative microscopic H or N involvement is a reliable prognostic sign, only extensive CT involvement of H or N predicts survival preoperatively. A better CT finding that predicts decreased survival preoperatively was extension out of the pancreatic head (especially S or A). Clinical methods of staging should use CT areas such as S, A, PV, and RP, and not H and N. PMID- 10365886 TI - The natural history of early recurrent carotid artery stenosis. AB - BACKGROUND: Early recurrent carotid stenosis, defined as greater than 50% stenosis within 2 years of a carotid endarterectomy (CEA), occurs in 4% to 19% of patients. These lesions are secondary to myointimal hyperplasia (MH). The natural history of these lesions has been examined prospectively, but the appropriate management of these lesions has not been clearly defined. The vascular surgery service at Madigan Army Medical Center (MAMC) has prospectively collected a cohort of patients with recurrent high-grade carotid stenoses following CEA to determine their natural history and define the ideal therapeutic approach for those lesions. METHODS: Patients undergoing CEA between January 1, 1993, and January 1, 1997, at a single tertiary care institution were followed prospectively with postoperative carotid duplexes at 3-month intervals for the first year and then every 6 months for a year and then annually thereafter. Data were collected regarding patient demographics, type of carotid closure, neurologic morbidity, and death. These results were compared with accepted rates in the literature. Discrete variables were tested for significance by chi-square analysis and Fisher's exact test. A P value less than or equal to 0.05 was considered significant. RESULTS: One hundred and seventy-four (174) patients with 181 operative sites were evaluated. Fourteen patients with 17 sites (9%) had recurrent stenosis. Twelve patients with 14 sites (7%) had stenoses of 50% to 79%. All were asymptomatic. Two patients with 3 sites (2%) had stenoses greater than 80%. Two sites were managed operatively because of neurologic symptoms or preocclusive nature and one remains asymptomatic and stable on serial duplex imaging. All lesions were present at 6 months and those in the 50% to 79% category did not progress in follow-up. Recurrent carotid stenosis occurred to a significantly higher degree in women (women 11 of 60 18.3% versus men 6 of 114 5.3%; P = 0.25), primary closure versus patch angioplasty (primary 6 of 22 27.3% versus patch 11 of 159 6.9%; P = 0.01), and dacron versus polytetrafluoroethylene (PTFE) patch angioplasty (dacron 7 of 36 19.4% versus PTFE 2 of 100 2.0%; P = 0.02). CONCLUSION: Early recurrent stenosis (50% to 79%) is a benign lesion. Patch angioplasty is preferred over primary closure. Dacron patches had a significantly higher rate of recurrent stenosis when compared with PTFE patches. Women undergoing CEA are more prone to recurrent stenosis. Postoperative duplex at 3 and 6 months will identify recurrent carotid stenosis (given a normal duplex prior to discharge following CEA). Moderate high-grade (50% to 79%) stenoses are benign. High-grade (80% to 99%) stenoses require individual management. PMID- 10365887 TI - Respiratory failure following talc pleurodesis. AB - BACKGROUND: Sterile talc is currently the agent of choice for pleurodesis. Its success rate is excellent, and talc is generally well tolerated. However, a recent experience with fulminant pneumonitis following talc pleurodesis prompted a review of our experience. METHODS: A retrospective review of patients undergoing talc pleurodesis at our institution between December 1993 and December 1997 was performed, documenting respiratory and other complications. Statistical analysis was performed using Student's t test and Pearson correlations. RESULTS: Seventy-eight patients received 89 talc pleurodesis procedures. Respiratory complications or death occurred in 33%; 9% of patients developed adult respiratory distress syndrome. There was no statistical difference in outcomes between patient groups, methods of application, or talc dosages utilized. CONCLUSIONS: This series revealed a significantly higher rate of serious complications than that reported in the current literature, without implicating a clear reason for these outcomes. Our data raise questions about the safety of talc pleurodesis. PMID- 10365888 TI - Surgical internet at a glance: volume IX. PMID- 10365889 TI - Clinical presentation and management of iatrogenic colon perforations. PMID- 10365890 TI - Diagnosis and management of adult intussusception. PMID- 10365891 TI - Weight loss has an independent beneficial effect on symptoms of gastro oesophageal reflux in patients who are overweight. AB - BACKGROUND: Weight loss is commonly recommended as part of first-line management of gastrooesophageal reflux disease (GORD) despite the paucity of published clinical trials. The aim of this study was to prospectively assess the independent effect of weight loss on reflux symptoms in overweight individuals with either normal endoscopic findings or grade-I oesophagitis. METHODS: Thirty four patients were recruited on the basis of a body mass index (BMI) of greater than 23 and symptoms of GORD for at least 6 months. All patients were advised to lose weight. Symptoms of gastro-oesophageal reflux (GOR) were scored, using a modified DeMeester questionnaire at 0, 6, and 26 weeks. Patients who were unable to stop taking all medication for control of symptoms were excluded from the study. Changes in weight and symptom score were analysed by using a paired t test. Correlation between change in weight and symptom score was assessed with the Pearson correlation test. RESULTS: Thirty-four patients were studied (18 men and 16 women) with a mean age of 65 years (range, 24-70 years). The mean weight at recruitment was 83.4 kg (standard deviation (s), 4.5 kg; BMI, 23.5 kg/m2 (s, 2.3 kg/m2). Twenty-seven patients (80% of the total) lost weight with a mean of 4.0 kg (P < 0.01) and improved by a mean reduction of 75% from the initial symptom score (P < 0.001). In nine patients the symptoms disappeared completely. Three patients gained weight and had a deterioration of their symptoms, whereas four patients gained weight but still improved their symptom score. There was a significant direct correlation between weight loss and symptom score (R = 0.548, P < 0.001). CONCLUSIONS: This study has shown a significant association between weight loss and improvement in symptoms of GOR. Patients who are overweight should be encouraged to lose weight as part of the first-line management. PMID- 10365892 TI - Rapid effect of lansoprazole on intragastric pH: a crossover comparison with omeprazole. AB - BACKGROUND: The time to onset of acid inhibition is considered an important factor when treating patients with reflux symptoms. This study was therefore designed to investigate the effect of 30 mg lansoprazole and 20 mg omeprazole on gastric pH after single-dose administration. METHODS: The study was of a randomized, open-label, single-dose and two-way crossover design with a washout period of at least 7 days in between. Fifteen healthy adult male and female subjects were included. The subjects were intubated with a pH catheter. Intragastric pH was measured every 4th sec for 10 min before and during 8 h after drug administration. Blood samples, for determination of plasma concentrations of lansoprazole and omeprazole, were obtained on 10 occasions during 6 h after drug administration. The area under the curve (AUC), the elimination halflife (t1/2), and the peak concentration (Cmax) of the two drugs were calculated. RESULTS: All subjects completed the study without major complications. The mean pH (0-8 h) after drug administration was 2.9 for lansoprazole and 2.0 for omeprazole (P = 0.0058). A pH of more than 4 was reached for the first time after 130 min with lansoprazole and after 250 min with omeprazole. AUC was 4919 +/- 2526 nmol/l x h for lansoprazole and 1352 +/- 1120 nmol/l x h for omeprazole. CONCLUSION: Single dose administration of 30 mg lansoprazole is followed by a rapid absorption of the drug and hence a more efficient acid suppression than after single-dose administration of 20 mg omeprazole in healthy volunteers. PMID- 10365893 TI - A close relationship between Helicobacter pylori infection and gastric xanthoma. AB - BACKGROUND: Although the pathogenesis of gastric xanthoma (GX) remains unclear, an association of GX with atrophic gastritis has been reported. Helicobacter pylori is closely related to atrophic gastritis. The aim of this study was to investigate the relationship among GX, H. pylori, and atrophic gastritis. METHODS: Sixty-seven patients with GX were assessed for H. pylori infection by serum anti-H. pylori IgG antibody, in addition to the rapid urease test, culture, and histologic examination using biopsy specimens of the antrum and corpus. The findings were compared with 67 age- and sex-matched control subjects without GX. The distribution of atrophic gastritis was assessed endoscopically. The severity of atrophic gastritis was determined endoscopically and histologically. Serum pepsinogen (PG) levels were also measured. Immunohistochemical staining of GX samples for H. pylori antigen was performed. H. pylori clinical isolates from patients with GX and controls were assessed for cagA by means of polymerase chain reaction. RESULTS: The prevalence of H. pylori was significantly higher in patients with GX than in controls (94% and 72%, respectively). A significantly more extensive atrophic gastritis was present in patients with GX, as determined endoscopically and histologically, than in controls. Serum PG-I levels and the PG I/PG-II ratio were significantly lower in the GX group than in the control group. H. pylori antigens were frequently identified in the cytoplasm of xanthoma cells in H. pylori-positive specimens of GX (54 of 63 specimens, 86%), whereas no immunoreactivity for H. pylori antigens was detected in H. pylori-negative specimens of GX. There was no significant difference in the positive rate of cagA between the two groups. CONCLUSIONS: Our results identified a close relationship among H. pylori infection, GX, and atrophic gastritis. A proportion of GXs may be provoked by H. pylori infection. PMID- 10365894 TI - Helicobacter pylori infection and risk of cardia cancer and non-cardia gastric cancer. A nested case-control study. AB - BACKGROUND: Helicobacter pylori infection is an established risk factor for gastric adenocarcinoma. Potential confounding by socioeconomic factors has not been adequately assessed, and the magnitude of the relative risk in relation to gastric subsites, morphologic subtypes, sex, age, and follow-up time need further study. METHODS: We conducted a serologic case-control study nested within the Norwegian JANUS cohort. Between 1972 and 1986 serum was collected from 101,601 subjects who were followed up with regard to cancer development through 1992. RESULTS: Among 208 gastric adenocarcinoma cases, we found a strong positive association between H. pylori infection and non-cardia gastric cancer (odds ratio (OR), 5.15; 95% confidence interval (CI), 2.83-9.37), and a statistically significant negative association with cardia cancer (OR, 0.40; 95% CI, 0.20 0.77). Adjustment for socioeconomic factors and smoking did not materially alter the effect estimates. The association between the infection and non-cardia cancer was stronger for tumors distal to the angulus and tended to be stronger in women than in men. The results were similar across Lauren morphologic subtypes. CONCLUSIONS: These results strengthen the evidence of H. pylori infection as a risk factor in non-cardia gastric cancer. A negative association with H. pylori infection was found for cardia cancer. PMID- 10365895 TI - Gastroduodenal mucosal vitamin-C levels in Helicobacter pylori infection. AB - BACKGROUND: Vitamin C is an important endogenous antioxidant, and epidemiologic evidence suggests that it may protect against the development of gastric cancer. We therefore determined mucosal vitamin-C levels in the stomach and duodenum of subjects with and without Helicobacter pylori infection. METHODS: The patients were 30 subjects undergoing routine gastroscopy for investigation of dyspepsia. High-performance liquid chromatography with electrochemical detection was used to determine mucosal ascorbic acid and total vitamin-C levels. RESULTS: In H. pylori negative subjects with normal gastroduodenal histology the antrum contained significantly higher levels of ascorbic acid and total vitamin C than the corpus or duodenum (P < 0.05). No significant changes were seen in gastric mucosal ascorbic acid or total vitamin-C levels in the presence of H. pylori infection and related inflammation. The presence of gastric atrophy did not affect mucosal ascorbic acid or total vitamin C levels. Duodenal ascorbic acid and total vitamin C levels did not change significantly in the presence of gastric H. pylori or duodenal inflammation. CONCLUSIONS: Although high levels of vitamin C are present in the gastroduodenal mucosa, these are not altered in the presence of H. pylori infection and inflammation. These observations suggest that the mucosal antioxidant potential of vitamin C is not impaired by H. pylori infection. PMID- 10365896 TI - A novel tablet-based 13C urea breath test for Helicobacter pylori with enhanced performance during acid suppression therapy. AB - BACKGROUND: The urea breath test (UBT) can still be improved in terms of user friendliness and accuracy during acid-suppression therapy. This study was designed to evaluate a novel, rapidly disintegrating 13C UBT tablet, which was supplemented with citric acid to facilitate diagnosis of Helicobacter pylori in the hypochlorhydric stomach. METHODS: The efficacy of a fasting 13C tablet-based UBT (TUBT) was compared with that of a standard 13C UBT (SUBT) during 40 min after dosing, and optimal sampling points were determined. The single-point TUBT was validated against a 'gold standard' (GS) including a TUBT, culture, histology, and a CLO test in 134 dyspeptic patients, and its optimal cut-off point was determined by means of a biometric method. In addition, 20 SUBT positive patients were randomized to perform either the TUBT or the SUBT after 7 days of omeprazole therapy (20 mg twice daily). RESULTS: Compared with a SUBT, the TUBT gave a quicker and wider separation between positive and negative results and an earlier optimal sampling point (10 versus 40 min). At 10 min the TUBT correctly classified 40 of 42 GS-positive subjects (sensitivity, 95%) and all of 92 GS-negative patients (specificity, 100%), and the optimal cut-off point was 1.8 delta per mil. Furthermore, when optimal sampling points were used, the TUBT (t = 10 min) proved to be more accurate than the SUBT (t = 40 min) during omeprazole treatment, correctly identifying all of 10 and 3 of 9 H. pylori infected patients, respectively. CONCLUSIONS: By supplying 13C urea and citric acid as a rapid-release tablet, it is possible to shorten the duration of the 13C UBT to 10 min, omit the test meal, and still maintain excellent accuracy, even during acid suppression therapy. PMID- 10365897 TI - Helicobacter pylori induces apoptosis in gastric mucosa through an upregulation of Bax expression in humans. AB - BACKGROUND: Maintenance of gastric mucosal integrity depends on the balance between cell loss due to apoptosis and cell proliferation. Helicobacter pylori induces apoptosis in gastric epithelial cells, but the regulation of this process has been little studied. The Bcl-2 proteins are the best-studied family of proteins involved in the mechanism of apoptotic death. Some members of this family, such as Bcl-2, inhibit apoptosis, whereas others, such as Bax, induce it. The present study was performed to determine the apoptosis rate and mRNA and protein expression for Bax and Bcl-2 in the gastric mucosa of duodenal ulcer (DU) patients with H. pylori infection before and after H. pylori eradication. We recruited 8 H. pylori-negative control subjects and 20 DU patients (H. pylori positive) given a 1-week triple therapy to eradicate H. pylori. The apoptosis was analyzed by means of terminal deoxyribonucleotide transferase-mediated digoxigenin-11-deoxyuridine triphosphate biotin nick-end labeling (TUNEL) staining, and the expression of mRNA for Bax and Bcl-2 by reverse transcription polymerase chain reaction (RT-PCR) and Southern blot. In all patients gastritis was assessed histologically on the basis of the Sydney classification, the presence of H. pylori, and analysis of cagA status. RESULTS: All 20 DU patients were H. pylori-positive, and 18 (90%) were CagA-positive. The apoptotic cells were infrequently identified in gastric surface epithelium by TUNEL histochemistry in H. pylori-negative controls. In DU patients infected with H. pylori, apoptotic cells were more numerous and seen deep in the gastric glands. The infection was associated with significantly upregulated expression of mRNA and protein for Bax and suppressed mRNA and protein expression for Bcl-2, as determined using RT-PCR and Western blot analysis. The Bax overexpression was significantly stronger in the antrum than in the corpus of H. pylori-infected patients. Four weeks after the eradication a marked decrease of neutrophil infiltration, an improvement of the grade of gastritis (mononuclear infiltration), and significant reduction in apoptosis rate were observed. After eradication the Bax mRNA expression was still at an increased level, whereas the Bcl-2 mRNA expression remained suppressed. CONCLUSIONS: 1) H. pylori induces apoptosis in the gastric epithelium, at least in part, due to an upregulation of proapoptotic Bax and downregulation of antiapoptotic Bcl-2, and 2) Bax mRNA and protein expression was higher in the antrum than in the corpus, and this was probably due to greater inflammatory changes observed in the antrum than in the corpus. PMID- 10365898 TI - H+/K+-adenosine triphosphatase mRNA in gastric fundic gland mucosa in patients infected with Helicobacter pylori. AB - BACKGROUND: How Helicobacter pylori infection affects gastric acid secretion has not been made clear. This study aimed to elucidate the effects of H. pylori infection on H+/K+-adenosine triphosphatase (ATPase) mRNA in gastric fundic gland mucosa. METHODS: Twenty patients with chronic gastritis and H. pylori infection were treated with lansoprazole and antibiotics. Before and 1 month after treatment gastroduodenoscopy was performed, and changes in the amount of H+/K+ ATPase mRNA in the fundic gland mucosa, gastric juice pH, and serum gastrin levels were determined. RESULTS: The amount of H+/ K+-ATPase mRNA in the fundic gland mucosa was increased in patients with eradication of H. pylori, in whom significant decreases in gastric juice pH and serum gastrin levels were observed. No significant changes were observed in patients without eradication of H. pylori. CONCLUSIONS: These results suggest that one of the mechanisms by which H. pylori infection suppresses acid secretion is by the inhibition of proton pump synthesis in parietal cells. PMID- 10365899 TI - Validation and value of an enzyme-linked immunosorbent assay for Helicobacter pylori in primary care. AB - BACKGROUND: Helicobacter pylori infection is related to gastric ulcer and carcinoma. In this study we validated the Pyloriset EIA-G New (a quantitative enzyme-linked immunosorbent assay) in a general practice population. The implications of eradication in case of a positive result were assessed. METHODS: One hundred and fifteen subsequent patients, enrolled in a randomized clinical trial evaluating the optimal strategy for treatment of dyspeptic patients in primary care, were included. Using biopsy as gold standard, we calculated the sensitivity and specificity of the test. RESULTS: The sensitivity and specificity of the test were 91% (95% confidence interval (CI) = 86% to 97%) and 78% (95% CI = 75% to 82%), respectively. Eradication of H. pylori would be indicated in 8 of 57 positive patients, since these actually had a peptic ulcer. In the other 49 patients eradication therapy would be unnecessary. CONCLUSION: The Pyloriset EIA G New is a reliable test in primary care. However, a serologic test-and-treat strategy in all dyspeptic patients cannot be recommended. PMID- 10365900 TI - Hyperglycaemia attenuates erythromycin-induced acceleration of solid-phase gastric emptying in idiopathic and diabetic gastroparesis. AB - BACKGROUND: Erythromycin has recently been found to be a gastrointestinal prokinetic agent in humans. Acute hyperglycaemia has been associated with delayed gastric emptying in both healthy controls and diabetic patients. Our aim was to investigate in gastroparetic patients (diabetics and idiopathics) whether hyperglycaemia, per se, reduces gastric motility during erythromycin-induced acceleration of gastric emptying of solids. METHODS: In 12 gastroparetic patients, 6 diabetics and 6 idiopathics, gastric emptying of solids was measured scintigraphically after giving placebo in normoglycaemia (5-8.9 mmol/l glucose) or 200 mg erythromycin lactobionate intravenously in normo- or hyperglycaemia (16 19 mmol/l glucose) induced by intravenous glucose infusion in random order on separate days. RESULTS: Erythromycin in normoglycaemia accelerated solids gastric emptying compared with placebo in all patients by abolishing the lag-phase duration and by decreasing the retained percentage of a meal in the stomach at 120 and 150 min (14.5% +/- 5.3% versus 88.4% +/- 10.6% and 3.5% +/- 2.1% versus 70.1% +/- 15.4%, respectively) (P < 0.001). The retained isotopic percentage in the stomach after erythromycin in induced hyperglycaemia compared with erythromycin in normoglycaemia, at 120 and 150 min, was increased (51.9% +/- 9.8% versus 14.5% +/- 5.3%, and 24.5% +/- 5.9% versus 3.5% +/- 2.1%, respectively) (P < 0.001) but was decreased in comparison with placebo (P < 0.001). A significantly increased percentage of isotope was retained in the stomach of the diabetic patients at 120 and 150 min, compared with the idiopathics, only after giving erythromycin in the hyperglycaemic condition (57.6% +/- 8.7% versus 46.1% +/- 7.6% (P = 0.036) and 27.8% +/- 5.7% versus 21.1 +/- 4.4% (P = 0.040), respectively). CONCLUSIONS: Hyperglycaemia attenuates erythromycin-induced acceleration of solid-phase gastric emptying in idiopathic and diabetic gastroparesis and increases the retained isotopic meal in the stomach. Hyperglycaemia reduces gastric motility more in the diabetic patients with gastroparesis than in idiopathic patients. PMID- 10365901 TI - Impaired release of peptide YY in patients with proctocolectomy and ileal pouch anal anastomosis. AB - BACKGROUND: Peptide YY (PYY) is a gut hormone produced by endocrine cells in the distal small bowel, colon, and rectum. PYY inhibits upper gastrointestinal secretory and motor functions. The aim of this study was to determine whether basal and postprandial plasma PYY levels in patients with proctocolectomy and ileal pouch-anal anastomosis (IPAA) are reduced and to determine the relationship between plasma PYY and plasma cholecystokinin (CCK) levels. METHODS: Plasma concentrations of PYY and CCK were measured before and after ingestion of a standardized breakfast in 14 IPAA patients and in 12 healthy control subjects. RESULTS: Basal PYY was slightly lower in the IPAA patients than in the controls (8.3 +/- 0.3 versus 9.3 +/- 1.1 pM; not significant). Ingestion of the meal induced a small but significant increase of PYY to a maximum of 10.9 +/- 0.9 pM in patients. Integrated postprandial PYY was markedly reduced in patients when compared with the controls (1725 +/- 66 pM*180min versus 3194 +/- 480 pM*180 min; P < 0.005). Plasma PYY concentrations were inversely correlated with plasma CCK concentrations in the 2nd and 3rd h after the meal (r = -0.86; P = 0.0001). CONCLUSION: PYY release in response to meal ingestion is markedly reduced but not completely absent in patients with proctocolectomy and ileal pouch-anal anastomosis. The inverse relationship between circulating PYY and CCK in the late postprandial phase is compatible with a negative feedback regulation of CCK release by endogenous PYY. PMID- 10365902 TI - Bowel problems in patients with systemic sclerosis. AB - BACKGROUND: The impact of systemic sclerosis on bowel function is still unknown. The aim of this study was therefore to assess the frequency and severity of colorectal problems among patients with systemic sclerosis and to determine whether these problems are associated with age, gender, type of systemic sclerosis, or time since diagnosis. METHODS: A detailed questionnaire describing diarrhoea, constipation, obstructed defecation, faecal incontinence, bowel habits, social activities, and quality of life was sent to 96 consecutive patients with systemic sclerosis. RESULTS: Among 83 respondents (86%) 16% did not have a normal desire to defecate, 18% regularly needed digital stimulation or evacuation of the rectum, and 38% had faecal incontinence. Most patients (79%) had episodes of diarrhoea, and 38% had this once or more each month. Overall, 20% reported that colorectal dysfunction caused some or a major restriction of social activities or the quality of life. CONCLUSIONS: Colorectal dysfunction is very common among patients with systemic sclerosis, often restricting social activities and the quality of life. Therefore, further studies of colorectal pathophysiology in patients with systemic sclerosis are needed. PMID- 10365903 TI - Population-based surveillance by colonoscopy: effect on the incidence of colorectal cancer. Telemark Polyp Study I. AB - BACKGROUND: Most cases of colorectal cancer (CRC) develop from adenomas. Polypectomy is believed to reduce the incidence of CRC, but this effect has never been explored in prospective controlled studies. The aim of the present study was to evaluate the effect of polypectomy on colorectal cancer incidence in a population-based screening program. METHODS: In 1983, 400 men and women aged 50 59 years were randomly drawn from the population registry of Telemark, Norway. They were offered a flexible sigmoidoscopy and, if polyps were found, a full colonoscopy with polypectomy and follow-up colonoscopies in 1985 and 1989. A control group of 399 individuals was drawn from the same registry. In 1996 both groups (age, 63-72 years) were invited to have a colonoscopic examination. Hospital files and the files of The Norwegian Cancer Registry were searched to register any cases of CRC in the period 1983-96. RESULTS: At screening endoscopy 324 (81%) individuals attended in 1983 and 451 (71%) in 1996. From 1983 to 1996, altogether 10 individuals in the control group and 2 in the screening group were registered to have developed CRC (relative risk, 0.2; 95% confidence interval (CI), 0.03-0.95; P = 0.02). A higher overall mortality was observed in the screening group, with 55 (14%) deaths, compared with 35 (9%) in the control group (relative risk, 1.57; 95% CI, 1.03-2.4; P = 0.03). CONCLUSION: Endoscopic screening examination with polypectomy and follow-up was shown to reduce the incidence of CRC in a Norwegian normal population. The possible effect of screening on overall mortality should be addressed in larger studies. PMID- 10365904 TI - The relationship between in vivo emptying of the gallbladder, biliary pain, and in vitro contractility of the gallbladder in patients with gallstones: is biliary colic muscular in origin? AB - BACKGROUND: This study sought to determine whether there is a positive correlation between gallbladder emptying, biliary pain, and in vitro contractility. METHODS: Ultrasound measurements were carried out on 25 gallstone patients. The response of gallbladder strips to 1.75*10(-11) to 5.25*10(-7) M cholecystokinin-8 was recorded. In a second study 23 patients filled in pain questionnaires, and in vitro studies were again carried out. RESULTS: Of five patients with no gallbladder emptying, four had in vitro contraction. Overall, a significant, positive linear correlation was found (P < 0.0001). In the second study in vitro contractility showed a positive linear correlation with pain. CONCLUSION: Gallbladder emptying correlates with contractility. However, since most 'non-contractors' can contract, we suggest the term 'non-emptying' or 'emptying' to describe gallbladder dynamics. The positive correlation between pain and contractility suggests that biliary pain has a muscular component. PMID- 10365905 TI - Carbohydrate 19-9 antigen as a marker of non-malignant hepatocytic ductular transformation in patients with acute liver failure. A comparison with alpha fetoprotein and carcinoembryonic antigen. AB - BACKGROUND: We have observed increased serum tumor markers, especially carbohydrate antigen 19-9 (CA 19-9) levels, in patients with acute liver failure (ALF) being evaluated for liver transplantation, raising the question of potential malignancy. In chronic liver disease increased serum alpha-fetoprotein (AFP) may be a sign of liver regeneration, but little is known of these markers in ALF. The aim of this study was to evaluate the causes of overexpression of tumor markers in patients with non-malignant ALF. METHODS: The serum AFP, carcinoembryonic antigen (CEA), and CA 19-9 levels were compared with the liver function tests in 33 patients with acute liver failure and in 78 patients with chronic non-malignant liver disease being evaluated for liver transplantation. Immunohistochemical stainings of the tumor markers were performed on explanted liver specimens. RESULTS: The AFP (1-218 U/ml) and CA 19-9 (10-6520 U/ml) levels were significantly higher in the patients with ALF than in the patients with chronic liver disease (P < 0.01). The AFP and CA 19-9 values also correlated with the total serum bilirubin level. In the patients with ALF the immunohistochemical staining for CA 19-9 was highly positive in periportal transformed ductular hepatocytes and correlated positively with the serum CA 19-9 values (P < 0.001). The stainings for AFP or CEA showed no or only slight positivity in the patients with increased serum values of the tumor markers. CONCLUSIONS: In patients with ALF increased serum levels of CA 19-9 reflect the amount of transformed ductular hepatocytes without any evidence of malignancy. Increased CA 19-9 values should not be the cause of delay when an ALF patient needs an urgent liver transplantation. PMID- 10365906 TI - Detection of hepatitis-C virus and hepatitis-C virus replication in hepatocellular carcinoma by in situ hybridization. AB - BACKGROUND: Although hepatitis C virus (HCV) is a major causative agent of hepatocellular carcinoma (HCC), the role of this virus in carcinogenesis is not fully understood. METHODS: We studied HCV infection and replication by detecting plus- and minus-strand HCV RNA by in situ hybridization (ISH) in surgically resected HCCs and adjacent non-cancerous tissue obtained from 15 anti-HCV antibody-positive patients with HCC. RESULTS: Plus-strand HCV RNA was found in 9 of 15 HCCs. Both plus- and minus-strand HCV RNA were detected in four of these nine patients. In non-cancerous tissues obtained adjacent to the HCC, plus-strand HCV RNA was found in 10 of 15 patients, whereas both plus- and minus-strand HCV RNA were detected in 7 of these 10 patients. The degree of staining by ISH did not correlate with the differentiation of the HCC, histologic classification of the non-cancerous tissue, serum HCV RNA levels, or serum transaminase levels. CONCLUSIONS: HCV can be found in and replicates in both hepatoma cells and in non cancerous hepatocytes in anti-HCV antibody-positive patients with HCC. The direct effects of HCV viral gene products on normal hepatocytes in the development of HCC require additional study. PMID- 10365907 TI - Feline model of chronic obstructive pancreatitis: effects of acute pancreatic duct decompression on blood flow and interstitial pH. AB - BACKGROUND: The mechanism by which duct decompression (DD) relieves pain in patients with chronic pancreatitis (CP) is unknown. CP is associated with increased tissue pressure (IP), low pancreatic microvascular blood flow (PMBF), and interstitial pH (pH(I)). The aims of this study were to examine the effects of acute DD on PMBF, increased IP, and pH(I) in cats with CP. METHODS: The main pancreatic duct was partially obstructed. At 6 weeks PMBF (ml/min/100g H2 gas clearance), IP (mmHg needle electrode), and pH(I) (microelectrode) were measured before and after secretin stimulation. The duct was then opened, and the studies were repeated. RESULTS: PMBF normally increased with secretin stimulation (118 +/ 20 versus 271 +/- 52, P < 0.05). IP was unaltered, and pH(I) decreased as hydrogen ions produced during bicarbonate secretion were dissipated (7.41 +/- 0.01 versus 7.38 +/- 0.01, P < 0.05). In CP, basal PMBF was lower than normal (51 +/- 6 versus 118 +/- 20, P < 0.05) and decreased with stimulation (51 +/- 3.5 versus 31 +/- 3.5, P < 0.05). Basal pancreatic IP was increased (3.47 +/- 0.7 versus 0.05 +/- 0.3, P < 0.05) and increased further with secretory stimulation (3.47 +/- 0.7 versus 4.41 +/- 0.7, P < 0.05) (a compartment syndrome). The low basal pancreatic pH(I) (7.23 +/- 0.02) did not change with secretin stimulation, since bicarbonate secretion was minimal. DD decreased IP (3.66 +/- 0.5 versus 2.81 +/- 0.5, P < 0.05) and increased PMBF (50 +/- 6 versus 79 +/- 6, P < 0.05) and pH(I) (7.24 +/- 0.02 versus 7.34 +/- 0.02, P < 0.05). The normal increase in PMBF after stimulation was restored (79 +/- 6 versus 218 +/- 54, P < 0.05). pH(I) now increased with stimulation (7.34 +/- 0.002 versus 7.37 +/- 0.002, P < 0.05), perhaps due to the marked hyperaemic response. CONCLUSIONS: DD acutely restored basal and stimulated PMBF and IP towards normal. Basal pancreatic pH(I) also improved and reflects the underlying ischaemia. PMID- 10365908 TI - Natural killer-like T-cell lymphoma of the stomach. AB - We report the case of a 69-year-old white woman who developed a natural killer (NK)-like T-cell lymphoma involving primarily the stomach. The tumour consisted of large and pleomorphic lymphocytes infiltrating the gastric mucosa. Immunohistochemistry performed on paraffin sections showed the neoplastic cells to be CD3+, CD5-, CD8-, CD43+, CD45RO+, and CD57+. In addition, these cells also expressed HLA-DR, granzyme B, and, to a lesser extent, the CD30 activation marker. No pathologic features suggesting Helicobacter pylori, Epstein-Barr virus infection, or lymphocytic gastritis were found within adjacent normal mucosa. The patient had no previous history of coeliac disease, and her serology for H. pylori was negative. Since lymphomas are usually considered the neoplastic counterpart of normal lymphocytic subsets, it is possible that in this case the tumour cells originate from a distinct cytotoxic T-cell population normally present within the gastric mucosa. The pathogenesis of this highly unusual neoplasm, however, remains a mystery. PMID- 10365909 TI - Effects of fixation on RNA extraction and amplification from laser capture microdissected tissue. AB - One of the key end points for understanding the molecular basis of carcinogenesis is the quantitation of gene expression in specific cell populations. Microdissection techniques allow extraction of morphologically distinct cells for molecular analysis. A recent advance in microdissection uses the PixCell laser capture microdissection (LCM) system, which allows for precise removal of pure cell populations from morphologically preserved tissue sections. The objective of this study was to determine the optimal fixation protocol for analyzing RNA from tissue samples using LCM. Optimal fixation must provide acceptable morphology, allow proper laser capture of selected cells, and preserve the integrity of mRNA. We evaluated the effects of both cross-linking and precipitive-type fixatives on frozen and paraffin-embedded mouse liver tissue. For assessment of the quality of the mRNA in LCM samples generated from various fixed tissues, reverse transcription-polymerase chain reaction (RT-PCR)-amplified mouse liver beta2 microglobulin mRNA was detected with ethidium bromide. We also examined mouse glyceraldehyde-3-phosphate-dehydrogenase by using the fluorogenic TaqMan system for real-time quantitative detection of RT-PCR products. Frozen tissues yielded more RT-PCR product than did paraffin-embedded tissues. In both frozen and paraffin-embedded tissues, differences were observed between the fixatives. Precipitive fixatives, such as ethanol and acetone, consistently produced more RT PCR amplification product than did cross-linking fixatives such as formalin. Optimal fixation protocols for LCM analysis will facilitate the examination of gene expression in specific cell populations, accelerating investigations of the molecular differences responsible for the phenotypic changes observed during carcinogenesis. PMID- 10365910 TI - Cdkn2a encodes functional variation of p16INK4a but not p19ARF, which confers selection in mouse lung tumorigenesis. AB - The cyclin-dependent kinase inhibitor 2a (Cdkn2a) locus encodes two distinct tumor suppressors, p16INK4a and p19ARF, whose functions interrelate in the regulation of cell proliferation as key components of the retinoblastoma and p53 pathways, respectively. In many types of cancer, alterations of Cdkn2a abrogate the functions of both suppressors, implying that both are integral to the genesis of certain cancer types. While this has been observed in mouse lung adenocarcinogenesis, recent observations also suggested that naturally occurring variation at the Cdkn2a locus is probably operative in the development of these tumors. Firstly, two common haplotypes of mouse Cdkn2a have been identified, each of which encodes cosegregating variants of p16INK4a and p19ARF. The p16INK4a variants differ at amino acids 18 (histidine or proline) and 51 (valine or isoleucine), whereas the p19ARF variants differ only at amino acid 72 (histidine or arginine). Secondly, genetic resistance to lung tumor formation appears to segregate with one particular haplotype, which also is deleted preferentially in lung adenocarcinomas of Cdkn2a heterozygous mice. Here we attempt to explain these observations and to characterize further the roles of p16INK4 and p19ARF in mouse lung tumorigenesis by examining the function and expression of each of the variants of Cdkn2a. Functional analysis showed that the proline 18/isoleucine 51 p16INK4a variant was diminished in cdk6 binding, cdk6 inhibition and NIH/3T3 fibroblast growth suppression compared with the histidine 18/valine 51 variant, whereas both of the p19ARF variants suppressed growth with similar potencies. Also, the different alleles for p16INK4a and p19ARF were transcribed equally in the normal lungs of Cdkn2a heterozygotes, as determined by comparative reverse transcription-polymerase chain reaction-single-stranded conformation polymorphism analysis. These results indicate that strain-specific variation in p16INK4a function is exploited in mouse lung tumorigenesis and strongly implicate a role for p16INK4a in lung cancer predisposition and development. PMID- 10365911 TI - Thirteenth Aspen Cancer Conference: workshop on mechanisms of toxicity and carcinogenesis. PMID- 10365912 TI - Frequent Ha-ras mutations in murine skin and liver tumors induced by 7H dibenzo[c,g]carbazole. AB - 7H-dibenzo[c,g]carbazole (DBC) is a potent liver and skin carcinogen when topically applied to the back skin of mice. This compound is found in complex mixtures produced during incomplete combustion of fossil fuels as well as in wood and tobacco smoke. The objective of this study was to elucidate the mechanism of action of this compound by assessing the Ha-ras mutational spectra of skin and liver tumors induced in a mouse model system. Low doses (50 nmol) and high doses (100 nmol) of DBC were applied topically to the backs of Hsd:ICR(Br) mice twice weekly. No treatment and solvent application were used as controls. After the mice were killed, the skin and liver tumors were removed, DNA was isolated, and tumor DNA was screened for Ha-ras codon 12, 13, and 61 mutations by using an enriched polymerase chain reaction method. Mutations were confirmed by reverse cyclic dideoxy sequencing. No mutations were found in codons 12 and 13 of DBC induced tumors, whereas one acetone-control tumor had a codon 13 mutation. Sixty seven percent of skin tumors and 45% of liver tumors induced by high doses of DBC and 67% of skin tumors induced by low doses of DBC contained codon 61 mutations, whereas liver tumors induced by low doses of DBC did not. The codon 61 mutations were exclusively A:T-->T:A transversions within the second base (CAA-->CTA). These results indicate that DBC is a unique polycyclic aromatic hydrocarbon in that it induces both skin and liver tumors upon topical application and that the mutational spectra are the same in tumors from two target organs, skin and liver, yet different from tumors from a third target organ, lung. PMID- 10365913 TI - Mutant p53: epigenetic mutator of the T-cell receptor via induction of methylation. AB - The mechanism and effects of epigenetic alterations in human carcinogenesis are not well understood, except that cancers often have alterations in the methylation status of their genomes. Additionally, human cancers, including aggressive T-cell leukemias and lymphomas, have a high frequency of p53 mutations, particularly missense mutations, which raises the possibility of gain of-new-function proteins, but the new proteins' oncogenic functions are mechanistically ill-defined. To investigate the mechanisms behind the high prevalence of p53 tumor suppressor gene mutations in aggressive or relapsed T cell leukemias, we transfected Jurkat cells null for p53 protein with a temperature-sensitive p53 mutant. We showed that this mutant p53 abrogated expression of the T-cell antigen receptor (TCR) by affecting the methylation of an at least 20-kb region of DNA, 5'to the TCR beta-chain gene enhancer region, which includes TCRbetaC1 and betaC2. Expression of the TCR is restored when the temperature is reduced to 32 degrees C, at which temperature the mutant p53 regains wild-type function. The TCR, a common site of dysfunction in T-cell malignancies, is the principal signal transduction moiety controlling both T-cell activation and activation-induced apoptosis. These results suggest a new role for mutant p53-as an epigenetic mutator, bridging p53, methylation, and transcriptional silencing-and suggest novel mechanisms in immunosuppression and cancer progression. PMID- 10365914 TI - Enhancement of susceptibility to diverse skin tumor promoters by activation of the insulin-like growth factor-1 receptor in the epidermis of transgenic mice. AB - Insulin-like growth factor-1 (IGF-1) and its receptor are believed to play an important role in mitogenesis and neoplastic transformation. The purpose of this study was to further examine the role of IGF-1 during tumor promotion in mouse skin. HK1.IGF1 transgenic mice, which overexpress IGF-1 in epidermis via the human keratin 1 promoter, were previously shown to be hypersensitive to skin tumor promotion by 12-O-tetradecanoylphorbol-13-acetate (TPA). We examined these mice for their sensitivity to diverse classes of tumor-promoting agents. HK1.IGF 1 transgenic mice initiated with 7,12-dimethylbenz[a]anthracene were more sensitive to treatment with a wide variety of tumor promoters, including chrysarobin, okadaic acid, and benzoyl peroxide, which resulted in more rapid development of tumors and a dramatic increase in the number of tumors per mouse compared with corresponding non-transgenic mice treated with the same compounds. Histological analyses of skin from HK1.IGF-1 mice treated with various tumor promoters revealed that these mice were also more sensitive to the induction of epidermal hyperplasia and cell proliferation. Analysis of the IGF-1 receptor (IGF 1r) and epidermal growth factor (EGFr) in the epidermis of TPA-treated HK1.IGF-1 transgenic and non-transgenic mice revealed that both receptors were activated (hyperphosphorylated on tyrosine residues), and the level of activation was higher in transgenic mice. The mechanism for the increased sensitivity of HK1.IGF 1 mice to tumor promoters may involve cooperation between the IGF-1r and EGFr signaling pathways. Our data suggest that IGF-1r signaling may play an important role in the process of tumor promotion by diverse classes of tumor promoters. PMID- 10365915 TI - p53 transactivity during in vitro osteoblast differentiation in a rat osteosarcoma cell line. AB - We previously demonstrated a correlation between wild-type p53 expression and appearance of osteoblastic-specific differentiation characteristics, as evidenced by basal osteocalcin gene expression in a mouse osteosarcoma tumor. The study reported here further explored the possibility of p53's having a distinct transcription-activating role in bone differentiation, in addition to its proposed role in G1 arrest and apoptosis. ROS17/2.3 osteoblastic osteosarcoma cells were stably transfected with a plasmid containing wild-type p53 binding sequences fused to the chloramphenicol acetyltransferase reporter gene. These cells were used to determine the transactivating role of p53 in regulation of osteocalcin gene expression. We chose two conditions under which osteocalcin expression is known to be upregulated: exposure of osteoblastic cells to differentiation-promoting medium and to vitamin D3. Exposure of the transfected cells to differentiation-promoting medium produced an increase in p53 transactivating activity correlating with the appearance of osteocalcin expression after about 1 wk. Vitamin D3 treatment resulted in upregulation of osteocalcin activity without a corresponding change in p53 transactivation activity or expression. In separate experiments, we tested whether changes in osteocalcin expression accompanied changes in p53 activity under conditions of downregulation of cell proliferation mediated by inhibition of DNA synthesis. Hydroxyurea treatment was used to inhibit DNA synthesis and produce growth arrest in osteoblastic cells. Inhibition of osteoblast cell proliferation was associated with a fourfold increase in p53 transactivating activity and a transient increase in osteocalcin steady-state expression. These results demonstrated a close relationship between p53 and osteocalcin and suggested a regulatory role for wild type p53 in the control of basal osteocalcin gene expression in osteoblasts. PMID- 10365916 TI - Leukocyte common antigen-related tyrosine phosphatase receptor: increased expression and neuronal-type splicing in breast cancer cells and tissue. AB - The findings that protein tyrosine phosphatases (PTPs) regulate cell proliferation, response to growth factors, and cellular adhesion and the discovery that mutations in PTP genes are associated with breast cancer suggest that altered expression of PTPs contributes to the breast cancer cell phenotype. The leukocyte common antigen-related (LAR) PTP receptor is a prototype member of the class of PTP receptors containing cell adhesion domains. Full-length constitutively spliced LAR transcripts are expressed in breast and other tissues, whereas alternatively spliced isoforms are preferentially expressed in the nervous system. As a first step in evaluating the hypothesis that LAR-type PTPs influence breast cancer cell behavior, LAR expression and neuronal-type alternative splicing were examined in normal and breast cancer cell lines and tissues. Northern blot analysis demonstrated markedly increased LAR mRNA levels in breast cancer cell lines and tissues. Western blot analysis showed a greater than tenfold increase in LAR protein levels in breast cancer tissues. Reverse transcription-polymerase chain reaction was used to assess alternative splicing of extracellular and proximal membrane exons. Differential patterns of extracellular alternative splicing were found in normal versus carcinoma cell lines and tissues. Western blot analysis demonstrated increased levels of LAR protein isoforms encoded by alternatively spliced transcripts in breast cancer cell lines. This study is the first demonstration of increased LAR mRNA and LAR protein expression in breast cancer tissue and nontransformed cell lines and helps to elucidate the role of LAR in human breast cancer. The differential patterns of alternative splicing of LAR transcripts introduce LAR isoforms as candidate markers for future studies correlating differential gene expression and tumor behavior. PMID- 10365917 TI - Inhibition of focal contact formation in cells transformed by p185neu. AB - Signaling pathways mediated by adhesive molecules are tightly associated with cytoskeletal organization and cell growth regulation. Focal adhesion kinase (FAK) plays a prominent role in the adhesion signaling pathway through its tyrosine kinase activity and protein-protein interaction with other signaling molecules, including src, paxillin, and p130CAS, and other proteins. We explored the roles of these signaling molecules in the transformation of B104-1-1 cells, an NIH/3T3 derived cell line transformed by activated rat p185neu. The cytoskeletal organization of the p185neu-transformed cells was disrupted, and their morphology was dramatically altered. FAK, paxillin, and p130CAS appeared to be tyrosine phosphorylated in both NIH/3T3 and B104-1-1. However, the phosphorylation levels of paxillin and p130CAS were lower in B104-1-1 cells than in NIH/3T3 cells. Surprisingly, the association between FAK and paxillin was enhanced in B104-1-1 cells, suggesting reorganization of protein-protein interaction modulated by protein phosphorylation. Our results showed that even though cellular transformation by src and neu has similar consequences, such as focal adhesion disassembly and increased metastasis potential, the molecular events underlying the signaling pathways can be dramatically different. PMID- 10365918 TI - The diagnostic value of cafe-au-lait macules. AB - Single cafe-au-lait macules (CALMs) are common in the pediatric population and in most children represent a normal finding. It is important to recognize whether the presence of multiple CALMs in a particular patient is normal or indicates an association with a multisystem disorder. This article addresses issues concerning the prevalence, genetics, and natural history of CALMs in the general population and reviews disorders in which CALMs are present as a characteristic trait. PMID- 10365919 TI - Suberythemogenic narrow-band UVB is markedly more effective than conventional UVB in treatment of psoriasis vulgaris. AB - BACKGROUND: Narrow-band UVB (NB-UVB) is a new phototherapy option for psoriasis. Action spectrum studies previously done with different UVB wavelengths suggest that suberythemogenic doses of NB-UVB could be highly effective in treating psoriasis vulgaris. Even so, no comparative studies with suberythemogenic doses of NB versus conventional UVB have been performed previously. OBJECTIVE: Our purpose was to compare conventional broad-band UVB (BB-UVB) with NB-UVB at suberythemogenic doses for the treatment of psoriasis vulgaris. METHODS: Eleven patients were treated using a split-body approach for 6 weeks on a three-times-a week basis. Outcomes were evaluated by means of Psoriasis Severity Index scores and quantitative histologic measures. RESULTS: We were able to induce clinical clearing in 81.8% of patients after NB-UVB, but in only 9.1% of patients after BB UVB (P < .01). Biopsy specimens obtained at the end of treatment revealed that keratin 16 staining was absent in 75% of patients on the NB side compared with none on the BB side, suggesting a reversal of regenerative epidermal hyperplasia by NB-UVB. CONCLUSION: NB-UVB is superior to UVB-BB in reversing psoriasis at suberythemogenic doses when given three times per week. This schedule was well tolerated by all patients. PMID- 10365920 TI - The epidermal phenotype during initiation of the psoriatic lesion in the symptomless margin of relapsing psoriasis. AB - BACKGROUND: The mature psoriatic lesion does not necessarily demonstrate changes relevant to early phases of the lesion. OBJECTIVE: In a model for relapsing psoriasis we examined the epidermal phenotype by means of a panel of immunohistochemical parameters: keratins 14 and 16, epidermal growth factor receptor (EGFR), Ki-67 antigen, and Tdt-mediated Unscheduled Nick End Labeling to detect apoptosis. METHODS: In 9 patients, we cleared psoriatic plaques by topical treatment with clobetasol-17-propionate under hydrocolloid occlusion. Relapse (defined as a clinical sum score > or = 6) was awaited. Biopsy specimens of the psoriatic lesion, the cleared skin, the relapsed plaque, and its clinically normal margin were assessed. RESULTS: Psoriasis recurred after 19+/-6 weeks (mean +/- SEM). During treatment all parameters improved considerably; however, the number of apoptotic cells was not affected. Ki-67 values decreased well below the normal range. At initial relapse, the symptomless skin adjacent to the relapsing lesion (margin) showed a marked expression of keratin 16 and EGFR. Ki-67 expression was increasing in the margin but was below values of the mature lesion. The localization of cycling cells in the first suprabasal layers was a remarkable feature. Keratin 14 expression was increased in the recurrent lesion itself, but not in the symptomless margin. CONCLUSION: Keratin 16 and EGFR expression are early phenomena in the evolution of the lesion, and they anticipate epidermal proliferation. The expression of keratin 14 follows overt epidermal hyperproliferation. The present observation in incipient psoriasis lends support to the hypothesis that the basal cell compartment does not have a primary involvement in the initiation of epidermal abnormalities in psoriasis, but that a coordinated sequence of events involving proliferation and differentiation markers in the first suprabasal layers of the epidermis could be the key to the pathogenesis of this puzzling disease. PMID- 10365921 TI - Antiperinuclear factor in psoriatic arthropathy. AB - BACKGROUND: Antiperinuclear factor (APF) is an autoantibody directed against (pro)-filaggrin molecules. OBJECTIVE: We evaluated whether APF determination is useful for the diagnosis of psoriatic arthritis (PA). METHODS: We determined APF titers in sera from patients with PA (n = 76), psoriasis vulgaris (n = 38), noninflammatory rheumatic diseases (NIRDs, n = 119), rheumatoid arthritis (RA, n = 159) both with negative (n = 36) and positive (n = 123) rheumatoid factor (RF) tests, as well as from 204 healthy controls. We used an indirect immunofluorescence test on epithelial cells from human buccal mucosa as a substrate. RESULTS: In patients with PA, 7.9% of the serum samples were APF positive. The incidence was greater than in healthy controls (1.0%; P < .01), similar to those with uncomplicated psoriasis (2.6%; P = NS) and NIRDs (4.0%; P = NS), and lower than in RF-negative (52.7%; P < .001) and RF-positive (90.2%; P < .001) patients with RA. Three APF-positive patients with PA had symmetric joint involvement and 3 had pustulotic arthroosteitis. CONCLUSION: The APF test may be useful in differentiating PA from RA, and APF may be specific for two PA subgroups. PMID- 10365922 TI - Prognostic factor analysis in mycosis fungoides/Sezary syndrome. AB - BACKGROUND: The influence of prognostic factors on survival in patients with mycosis fungoides/Sezary syndrome (MF/SS) is less well described than for other lymphomas. OBJECTIVE: Our purpose was to evaluate the prognostic value of diverse clinicopathologic and laboratory characteristics in patients with MF/SS. METHODS: All 115 patients with MF/SS seen at the Mycosis Fungoides clinic at M. D. Anderson Cancer Center during the study period who had slides available for pathologic review were analyzed. Univariate and multivariate methodologies were used. RESULTS: Age (> or = 60 years; P = .0002), advanced stage (P < 10(-5)), tumor (T3) stage disease (P < or = 10(-5)), lymphadenopathy (P = .006), bone marrow infiltration (P = .03), high lactate dehydrogenase (LDH; P = .0002), high beta2-microglobulin (> 2 mg/L; P = .009), and transformation to large-cell lymphoma (P = .004) were significant prognostic factors in the univariate analysis and correlated with a poorer survival. The outcome of patients staged as IIB was significantly worse than that of those staged as I or IIA or III (P < .001) and was comparable to that of the patients staged as IV (P = .8). In the multivariate analysis, the factors selected include stage (I to IIA and III vs IIB and IV; P < .0001), LDH (P = .006), and age (> or = 60 vs < 60 years; P = .02). The actuarial median survival of patients with advanced stage, high LDH, or age 60 years or more was 2.5 to 3.5 years, whereas that of patients without any of these parameters was more than 13 years. CONCLUSION: Our results suggest that patients with MF/SS who are staged as IIB or IV, who have a high LDH, or who are 60 years of age or older have an aggressive course and poor survival. PMID- 10365923 TI - The antiangiogenic agents TNP-470 and 2-methoxyestradiol inhibit the growth of angiosarcoma in mice. AB - BACKGROUND: Endothelial malignancies, such as angiosarcoma and hemangioendothelioma, are often resistant to chemotherapy and surgery, and may result in death. Improved means of therapy are needed for these disorders. OBJECTIVE: We wanted to determine whether angiosarcoma can be treated with angiogenesis inhibitors in mice. METHODS: Mice were inoculated with a cell line that gives rise to angiosarcoma and were treated with the angiogenesis inhibitors 2-methoxyestradiol and TNP-470. Response to therapy was monitored by measurement of tumors. RESULTS: TNP-470 caused an 84% reduction in tumor size, and 2 methoxyestradiol caused a 68% reduction in tumor size. CONCLUSION: Angiogenesis inhibitors are highly effective in treatment of angiosarcoma in mice. Clinical trials of these agents in humans with angiosarcoma and hemangioendothelioma are warranted. PMID- 10365925 TI - Nongenital dermatologic disease in HIV-infected women. AB - BACKGROUND: Dermatologic disease in HIV-infected women has not been adequately characterized. OBJECTIVE: The main purposes of this study were to characterize nongenital dermatologic disease in HIV-infected women and correlate these diagnoses with CD4 lymphocyte count to compare these findings with those in published reports of men. METHODS: This study was a retrospective chart review of female patients with dermatologic diagnoses followed up at an HIV clinic in New York City, seen by either a dermatologist (49 patients) and/or a primary care practitioner (114 patients). CD4 lymphocyte count was recorded if available within 6 months of diagnosis; mean CD4 count was calculated for all disorders with 5 or more diagnoses. RESULTS: Oropharyngeal candidiasis, drug eruption, dermatophytosis, rash (not otherwise specified), nongenital herpes simplex, herpes zoster, and seborrheic dermatitis were the most prevalent diagnoses made by primary care providers. Itchy red bump disease, acne, atopic dermatitis, xerosis, seborrheic dermatitis, nongenital warts, and molluscum contagiosum were the most prevalent diagnoses made by the dermatologist. Mean CD4 lymphocyte count was lowest in itchy red bump disease, nongenital warts, nongenital herpes simplex, xerosis, and drug eruptions. CONCLUSION: There appears to be no appreciable difference in the spectrum or prevalence of dermatologic disease in HIV-infected women versus HIV-infected men, except for a lower prevalence of Kaposi's sarcoma, oral hairy leukoplakia, and possibly onychomycosis in women. The degree of immunosuppression associated with various dermatoses in HIV infected women is similar to that in men, except perhaps for molluscum contagiosum, which may appear earlier in women. PMID- 10365924 TI - Finasteride in the treatment of men with frontal male pattern hair loss. AB - BACKGROUND: Finasteride, a specific inhibitor of type II 5alpha-reductase, decreases serum and scalp dihydrotestosterone and has been shown to be effective in men with vertex male pattern hair loss. OBJECTIVE: This study evaluated the efficacy of finasteride 1 mg/day in men with frontal (anterior/mid) scalp hair thinning. METHODS: This was a 1-year, double-blind, placebo-controlled study followed by a 1-year open extension. Efficacy was assessed by hair counts (1 cm2 circular area), patient and investigator assessments, and global photographic review. RESULTS: There was a significant increase in hair count in the frontal scalp of finasteride-treated patients (P < .001), as well as significant improvements in patient, investigator, and global photographic assessments. Efficacy was maintained or improved throughout the second year of the study. Finasteride was generally well tolerated. CONCLUSION: In men with hair loss in the anterior/mid area of the scalp, finasteride 1 mg/day slowed hair loss and increased hair growth. PMID- 10365926 TI - Underpants-pattern erythema: a previously unrecognized cutaneous manifestation of extramammary Paget's disease of the genitalia with advanced metastatic spread. AB - BACKGROUND: A previously unrecognized erythema was noted in 6 patients with extramammary Paget's disease (EMPD) of the genitalia with metastases. It was distinct from essential erythema of EMPD and has not been reported before. OBJECTIVE: Our purpose was to describe the clinical and histopathologic features of the erythema and clarify its pathogenesis and prognostic importance. METHODS: A review of the clinical records and histologic materials of 6 cases was made focusing on the clinical features of this unique erythema as well as the course and prognosis of the patients. RESULTS: The erythema first appeared in the inguinal region and extended centrifugally to an area normally covered by underpants. Histopathologic examination showed lymphatic infiltration by cancer cells of the skin. All of the patients with this "underpants-pattern erythema" died of the disease, and the mean survival period after its appearance was 13 months. CONCLUSION: Underpants-distribution erythema may be a sign of advanced stage EMPD. PMID- 10365927 TI - Mycophenolate mofetil: a new therapeutic option in the treatment of blistering autoimmune diseases. AB - BACKGROUND: Mycophenolate mofetil (MMF), an ester of mycophenolic acid (MPA), was approved by the Food and Drug Administration in 1995 and is currently primarily indicated for the prophylaxis of rejection in renal transplant patients. The drug seems also to be of value in the treatment of psoriasis and rheumatic arthritis. Recently there have been 6 reported cases of successful treatment of blistering autoimmune diseases with MMF in combination with high dose prednisone therapy. OBJECTIVE: On the basis of these reports we administered this new treatment regimen to several patients with blistering autoimmune diseases. Besides using a combination of MMF and high-dose prednisone we wanted to evaluate whether MMF monotherapy is also effective in the treatment of blistering autoimmune diseases. METHODS: We administered MMF to 5 patients who had severe pemphigus vulgaris or bullous pemphigoid. Two patients received MMF in combination with high-dose prednisone therapy and 3 patients received MMF monotherapy. To our knowledge, this is the first report of successful treatment of pemphigus vulgaris and bullous pemphigoid with MMF monotherapy. RESULTS: All patients were completely free of symptoms within 8 to 11 weeks of therapy. Patients who had received MMF monotherapy responded as well to treatment as those who received a combination of MMF and high-dose prednisone. CONCLUSION: Our experiences strongly suggest that MMF monotherapy may be effective for patients even with severe pemphigus vulgaris and bullous pemphigoid. In addition, MMF monotherapy, at least over the short term, offers the advantage of fewer side effects in comparison to immunosuppressive combination therapy and was well tolerated by our patients. PMID- 10365928 TI - A comparison of topical azelaic acid 20% cream and topical metronidazole 0.75% cream in the treatment of patients with papulopustular rosacea. AB - BACKGROUND: Although it is important for physicians to have sufficient clinical data on which to base treatment decisions, little comparative data exist regarding newer treatment modalities for rosacea. OBJECTIVE: The goal of the study was to compare the efficacy and safety of topical azelaic acid 20% cream and topical metronidazole 0.75% cream in the treatment of patients with papulopustular rosacea. Parameters of patient satisfaction to treatment were also assessed. METHODS: Forty patients with the clinical manifestation of symmetric facial rosacea were investigated in this single-center, double-blind, randomized, contralateral split-face comparison clinical trial. RESULTS: After 15 weeks of treatment, both azelaic acid and metronidazole induced significant, albeit equal reductions in the number of inflammatory lesions (pustules and papules). A significantly higher physician rating of global improvement was achieved with azelaic acid. Changes in the rosacea signs and symptoms of dryness, burning, telangiectasia, and itching were equal between treatments. A reduction in erythema tended toward significance with azelaic acid at week 15. A trace amount of stinging on application was noted with azelaic acid; however, such discomfort did not appear to concern patients because their overall impression of azelaic acid was superior to that of metronidazole. CONCLUSION: Azelaic acid 20% cream provides an effective and safe alternative to metronidazole 0.75% cream with the added benefit of increased patient satisfaction. PMID- 10365929 TI - Specific cutaneous infiltrates in patients with myelogenous leukemia: a clinicopathologic study of 26 patients with assessment of diagnostic criteria. AB - BACKGROUND: Few recent studies have analyzed the clinicopathologic features of specific cutaneous manifestations of myelogenous leukemia in a large number of patients. OBJECTIVE: We characterize the clinical and histopathologic spectrum of specific cutaneous manifestations in acute (AML) and chronic (CML) myelogenous leukemia, ascertain further diagnostic criteria, and examine current prognosis. METHODS: Thirty-six lesions of specific cutaneous infiltrates from 26 patients with myelogenous leukemia (AML: 17 patients; M:F = 1:2.4; mean age: 52.6 years; AML-French-American-British [FAB] classification subtypes:M1 = 1, M2 = 3, M4 = 8, M5 = 5. CML = 9 patients; M:F = 4.5:1; mean age: 60.6 years) were retrospectively collected for the study. RESULTS: Cutaneous manifestations presented as solitary or multiple reddish to violaceous papules, plaques, and nodules (17 lesions), or as a generalized erythematous maculopapular eruption (9 lesions). Concurrent extramedullary involvement in other peripheral sites (eg, gums, pharynx, orbits) was observed in 10 patients. Histopathologically, lesions revealed nodular/diffuse infiltrates, often with perivascular and periadnexal accentuation, sparing of the upper papillary dermis, and prominent single arraying of neoplastic cells between collagen bundles. Extension to the subcutis was noted in all deep biopsy specimens (26 lesions). Cytomorphologically, medium to large-sized mononuclear cells (myeloblasts and atypical myelocytes) predominated in AML-M1 and M2, whereas M4 and M5 mainly showed small, medium sized, or large mononuclear cells with slightly eosinophilic cytoplasm and indented, bi-lobular, or kidney-shaped nuclei (atypical monocytoid cells). In CML, either a variable mixture of mature and immature cells of the granulocytic series (myelocytes, metamyelocytes, eosinophilic metamyelocytes, and neutrophils) or a rather monomorphous infiltrate of mononuclear cells were found. Staining for naphthol AS-D chloroacetate-esterase (NASD) was positive in 24 of 36 lesions (66.6%; AML: 16; CML: 8). Immunohistochemical analysis on paraffin sections using a large panel of antibodies (16 lesions: AML: 13; CML: 3) showed strong reactivity for LCA (CD45), lysozyme, myeloperoxidase (MPD), LN2 (CD74), HLA-DR, and MT1 (CD43) in the majority of cases, and variable staining for monocyte/macrophage markers (KP1/CD68, PGM1/CD68, Mac387, Ki-M1p). The neuronal cell adhesion molecule (NCAM) marker CD56 was reactive in 2 cases of CML, but negative in all cases of AML. MIB1(Ki67) stained 20% to 80% of neoplastic cells. CD34, CD15, CD20, and CD3 were negative in all cases. No correlation between histochemical/immunohistochemical features with type of leukemia or FAB-subtype of AML was observed. All patients with CML and AML with adequate follow-up died within 24 months after onset of skin lesions (mean survival, AML: 7.6 months; CML: 9.4 months). CONCLUSION: Specific cutaneous lesions in AML and CML show distinctive clinicopathologic features that allow diagnosis in most cases. Immunohistochemistry on routinely fixed, paraffin-embedded tissue sections provides useful adjunctive information. Simultaneous expression of lysozyme, MPD, CD45, CD43, and CD74 militates in favor of a diagnosis of specific cutaneous infiltrate of myelogenous leukemia. Pitfalls in immunohistologic diagnosis mainly include lack of expression of some myeloid markers (lysozyme, MPD), and aberrant expression of T-cell markers (eg, CD45RO). Regardless of type of myelogenous leukemia, onset of specific skin manifestations correlates with an aggressive course and short survival. PMID- 10365931 TI - Surgical pearl: The high-energy pulsed carbon dioxide laser for immediate scar resurfacing. PMID- 10365930 TI - Five cases of calciphylaxis and a review of the literature. AB - Calciphylaxis is a rare phenomenon of cutaneous necrosis that typically occurs in association with renal failure and has a poor prognosis. We report 5 new cases of calciphylaxis that illustrate the important clinical and histopathologic features of the disease. All patients had end-stage renal failure at the time that purpuric plaques and nodules were noted; these subsequently progressed to necrotic ulcers with eschars. All skin biopsy specimens showed varying degrees of calcification of the medial layer of blood vessel walls in the dermis and subcutaneous fat. Neither the product of serum calcium and phosphorus concentrations nor parathyroid hormone levels correlated temporally with the clinical observations in every case, emphasizing the importance of clinical histopathologic correlation. Although certain features of calciphylaxis in humans resemble the animal model originally proposed, there are also some crucial differences. We review the pathogenesis, epidemiology, clinical and histopathologic features, and treatment of this disease. PMID- 10365932 TI - Protein-contact eczematous reaction to cornstarch in clothing. AB - Protein contact dermatitis is an eczematous reaction to antigens often associated with immediate hypersensitivity. A patient with a history of atopic eczema and multiple immediate sensitivities was seen for a persistent dermatitis of the face, hands, and clothing areas of the trunk and arms. Investigation showed a positive prick test to cornstarch, and avoidance of glove powder and starch in her clothing cleared what had been a recalcitrant problem. PMID- 10365933 TI - Phototherapy for atopic eczema with narrow-band UVB. AB - Management of atopic dermatitis has been less than satisfactory. Conventional therapy has not been particularly successful, and prolonged use of topical corticosteroids and systemic immunosuppressant drugs (eg, corticosteroids, cyclosporine, azathioprine) can result in severe cutaneous and systemic effects. We decided to evaluate the effect of UVB at 311 nm to treat 5 patients with moderate to severe atopic dermatitis. In each patient a mean cumulative dose of 9.2 J/cm2 was applied over a mean of 19 irradiations. Narrow-band UVB notably reduced atopic dermatitis after 3 weeks in all patients. PMID- 10365934 TI - Readability of skin cancer prevention brochures targeting parents of young children. AB - Long-term exposure to UV radiation and severe sunburns increase one's risk of experiencing malignant melanoma later in life, so parents need to be informed about how to protect children from overexposure to the sun. We attempted to determine readability of skin cancer brochures targeted toward parents of young children. SMOG and FOG readability formulas were applied to 8 brochures published by the American Cancer Society, Skin Cancer Foundation, American Academy of Dermatology, and the Anti-Cancer Council. Readability levels of the brochures ranged between the 8th and 12th grade levels. To reduce the incidence of skin cancer, sun-safe knowledge and behavior should start in childhood. Pediatricians need access to brochures written at readability levels for the average parent. Skin cancer brochures written at the eighth or ninth grade comprehension levels may allow pediatricians to educate more parents about the importance of skin cancer prevention in childhood. PMID- 10365935 TI - Tretinoin in the removal of eyeliner tattoo. AB - Eyelid tattooing is a commonly performed procedure. For at least 100 years, it has been performed by medical and nonmedical professionals. Complications can occur; the main one is improperly placed pigment. To date, the most frequently reported methods to remove eyeliner tattoos have been laser treatments or surgical correction. We observed a case in which tretinoin was used successfully in the removal of an eyelid tattoo. PMID- 10365936 TI - Cutaneous New World leishmaniasis-sporotrichosis coinfection: report of 3 cases. AB - Three cases of coinfection with Leishmania and Sporothrix spp in the same lesion are described. The patients had ulcers with erythematous borders and regional lymphadenopathy. The diagnosis of leishmaniasis was accomplished by direct visualization of the amastigotes or culture of the promastigotes, or both. The diagnosis of sporotrichosis was proved in two cases by culture of Sporothrix schenckii and by the histopathologic features in one case. All patients had a positive sporotrichin test. Two patients responded successfully to oral potassium iodide. One patient received oral itraconazole 100 mg/day because of intolerance to iodides and was cured. To our knowledge coinfection with Leishmania and Sporothrix spp has not been reported. The use of empirical treatments for leishmaniasis such as poultices or puncturing of the lesion with thorns or woods splinters might introduce Sporothrix and explain the coinfection. PMID- 10365937 TI - Treatment of cutaneous lymphoid hyperplasia with thalidomide: report of two cases. AB - Cutaneous benign lymphoid hyperplasia is a B-cell pseudolymphoma of unknown origin. The most favored sites of involvement include the face. We report two cases involving the nose that showed complete and stable regression after a 2 month treatment course with thalidomide. PMID- 10365938 TI - The future ain't what it was and are we ready for it? PMID- 10365939 TI - Prescribing and cost analysis of treatments of cutaneous fungal infections. PMID- 10365940 TI - Phenylalanine with UVA for the treatment of vitiligo needs more testing for possible side effects. PMID- 10365942 TI - Policy on papers' contributors. PMID- 10365941 TI - Cyclosporine-associated lymphoma. PMID- 10365943 TI - Proceed with caution, says UK report on ethics of GM foods. PMID- 10365944 TI - US Senate gets tough on animal activists. PMID- 10365945 TI - 'Frontiers' grants count cost of success. PMID- 10365946 TI - Cautionary tale on safety of GM crops. PMID- 10365947 TI - Plant DNA patents in the hands of a few. PMID- 10365948 TI - Papers should spell out authors' roles. PMID- 10365949 TI - Deer destiny determined by density. PMID- 10365950 TI - Apoptosis. Key to the mitochondrial gate. PMID- 10365951 TI - Retroviruses. Closing the joint. PMID- 10365953 TI - Evolutionary neurobiology. The neocortex comes together. PMID- 10365952 TI - Quantum enzymology. Tunnel vision. PMID- 10365954 TI - Release from inhibition reveals the visual past. PMID- 10365955 TI - Tying a molecular knot with optical tweezers. AB - Filamentous structures are abundant in cells. Relatively rigid filaments, such as microtubules and actin, serve as intracellular scaffolds that support movement and force, and their mechanical properties are crucial to their function in the cell. Some aspects of the behaviour of DNA, meanwhile, depend critically on its flexibility-for example, DNA-binding proteins can induce sharp bends in the helix. The mechanical characterization of such filaments has generally been conducted without controlling the filament shape, by the observation of thermal motions or of the response to external forces or flows. Controlled buckling of a microtubule has been reported, but the analysis of the buckled shape was complicated. Here we report the continuous control of the radius of curvature of a molecular strand by tying a knot in it, using optical tweezers to manipulate the strand's ends. We find that actin filaments break at the knot when the knot diameter falls below 0.4 microm. The pulling force at breakage is around 1 pN, two orders of magnitude smaller than the tensile stress of a straight filament. The flexural rigidity of the filament remained unchanged down to this diameter. We have also knotted a single DNA molecule, opening up the possibility of studying curvature-dependent interactions with associated proteins. We find that the knotted DNA is stronger than actin. PMID- 10365956 TI - Population density affects sex ratio variation in red deer. AB - Many mammal populations show significant deviations from an equal sex ratio at birth, but these effects are notoriously inconsistent. This may be because more than one mechanism affects the sex ratio and the action of these mechanisms depends on environmental conditions. Here we show that the adaptive relationship between maternal dominance and offspring sex ratio previously demonstrated in red deer (Cervus elaphus), where dominant females produced more males, disappeared at high population density. The proportion of males born each year declined with increasing population density and with winter rainfall, both of which are environmental variables associated with nutritional stress during pregnancy. These changes in the sex ratio corresponded to reductions in fecundity, suggesting that they were caused by differential fetal loss. In contrast, the earlier association with maternal dominance is presumed to have been generated pre-implantation. The effects of one source of variation superseded the other within about two generations. Comparison with other ungulate studies indicates that positive associations between maternal quality and the proportion of male offspring born have only been documented in populations below carrying capacity. PMID- 10365957 TI - Multifractality in human heartbeat dynamics. AB - There is evidence that physiological signals under healthy conditions may have a fractal temporal structure. Here we investigate the possibility that time series generated by certain physiological control systems may be members of a special class of complex processes, termed multifractal, which require a large number of exponents to characterize their scaling properties. We report on evidence for multifractality in a biological dynamical system, the healthy human heartbeat, and show that the multifractal character and nonlinear properties of the healthy heart rate are encoded in the Fourier phases. We uncover a loss of multifractality for a life-threatening condition, congestive heart failure. PMID- 10365958 TI - Decrystallization of adult birdsong by perturbation of auditory feedback. AB - Young birds learn to sing by using auditory feedback to compare their own vocalizations to a memorized or innate song pattern; if they are deafened as juveniles, they will not develop normal songs. The completion of song development is called crystallization. After this stage, song shows little variation in its temporal or spectral properties. However, the mechanisms underlying this stability are largely unknown. Here we present evidence that auditory feedback is actively used in adulthood to maintain the stability of song structure. We found that perturbing auditory feedback during singing in adult zebra finches caused their song to deteriorate slowly. This 'decrystallization' consisted of a marked loss of the spectral and temporal stereotypy seen in crystallized song, including stuttering, creation, deletion and distortion of song syllables. After normal feedback was restored, these deviations gradually disappeared and the original song was recovered. Thus, adult birds that do not learn new songs nevertheless retain a significant amount of plasticity in the brain. PMID- 10365959 TI - Neuronal correlates of parametric working memory in the prefrontal cortex. AB - Humans and monkeys have similar abilities to discriminate the difference in frequency between two mechanical vibrations applied sequentially to the fingertips. A key component of this sensory task is that the second stimulus is compared with the trace left by the first (base) stimulus, which must involve working memory. Where and how is this trace held in the brain? This question was investigated by recording from single neurons in the prefrontal cortex of monkeys while they performed the somatosensory discrimination task. Here we describe neurons in the inferior convexity of the prefrontal cortex whose discharge rates varied, during the delay period between the two stimuli, as a monotonic function of the base stimulus frequency. We describe this as 'monotonic stimulus encoding', and we suggest that the result may generalize: monotonic stimulus encoding may be the basic representation of one-dimensional sensory stimulus quantities in working memory. Thus we predict that other behavioural tasks that require ordinal comparisons between scalar analogue stimuli would give rise to monotonic responses similar to those reported here. PMID- 10365960 TI - Developmental basis of limblessness and axial patterning in snakes. AB - The evolution of snakes involved major changes in vertebrate body plan organization, but the developmental basis of those changes is unknown. The python axial skeleton consists of hundreds of similar vertebrae, forelimbs are absent and hindlimbs are severely reduced. Combined limb loss and trunk elongation is found in many vertebrate taxa, suggesting that these changes may be linked by a common developmental mechanism. Here we show that Hox gene expression domains are expanded along the body axis in python embryos, and that this can account for both the absence of forelimbs and the expansion of thoracic identity in the axial skeleton. Hindlimb buds are initiated, but apical-ridge and polarizing-region signalling pathways that are normally required for limb development are not activated. Leg bud outgrowth and signalling by Sonic hedgehog in pythons can be rescued by application of fibroblast growth factor or by recombination with chick apical ridge. The failure to activate these signalling pathways during normal python development may also stem from changes in Hox gene expression that occurred early in snake evolution. PMID- 10365961 TI - The MAPK kinase Pek1 acts as a phosphorylation-dependent molecular switch. AB - The mitogen-activated protein kinase (MAPK) pathway is a highly conserved eukaryotic signalling cascade that converts extracellular signals into various outputs, such as cell growth and differentiation. MAPK is phosphorylated and activated by a specific MAPK kinase (MAPKK): MAPKK is therefore considered to be an activating regulator of MAPK. Pmk1 is a MAPK that regulates cell integrity and which, with calcineurin phosphatase, antagonizes chloride homeostasis in fission yeast. We have now identified Pek1, a MAPKK for Pmk1 MAPK. We show here that Pek1, in its unphosphorylated form, acts as a potent negative regulator of Pmk1 MAPK signalling. Mkh1, an upstream MAPKK kinase (MAPKKK), converts Pek1 from being an inhibitor to an activator. Our results indicate that Pek1 has a dual stimulatory and inhibitory function which depends on its phosphorylation state. This switch-like mechanism could contribute to the all-or-none physiological response mediated by the MAPK signalling pathway. PMID- 10365962 TI - Bcl-2 family proteins regulate the release of apoptogenic cytochrome c by the mitochondrial channel VDAC. AB - During transduction of an apoptotic (death) signal into the cell, there is an alteration in the permeability of the membranes of the cell's mitochondria, which causes the translocation of the apoptogenic protein cytochrome c into the cytoplasm, which in turn activates death-driving proteolytic proteins known as caspases. The Bcl-2 family of proteins, whose members may be anti-apoptotic or pro-apoptotic, regulates cell death by controlling this mitochondrial membrane permeability during apoptosis, but how that is achieved is unclear. Here we create liposomes that carry the mitochondrial porin channel (also called the voltage-dependent anion channel, or VDAC) to show that the recombinant pro apoptotic proteins Bax and Bak accelerate the opening of VDAC, whereas the anti apoptotic protein Bcl-x(L) closes VDAC by binding to it directly. Bax and Bak allow cytochrome c to pass through VDAC out of liposomes, but passage is prevented by Bcl-x(L). In agreement with this, VDAC1-deficient mitochondria from a mutant yeast did not exhibit a Bax/Bak-induced loss in membrane potential and cytochrome c release, both of which were inhibited by Bcl-x(L). Our results indicate that the Bcl-2 family of proteins bind to the VDAC in order to regulate the mitochondrial membrane potential and the release of cytochrome c during apoptosis. PMID- 10365963 TI - Interaction of E1 and hSNF5 proteins stimulates replication of human papillomavirus DNA. AB - Mammalian viruses often use components of the host's cellular DNA replication machinery to carry out replication of their genomes, which enables these viruses to be used as tools for characterizing factors that are involved in cellular DNA replication. The human papillomavirus (HPV) E1 protein is essential for replication of the virus DNA. Here we identify the cellular factor that participates in viral DNA replication by using a two-hybrid assay in the yeast Saccharomyces cerevisiae and E1 protein as bait. Using this assay, we isolated Inil/hSNF5, a component of the SWI/SNF complex which facilitates transcription by altering the structure of chromatin. In vitro binding and immunoprecipitation confirmed that E1 interacts directly with Ini1/hSNF5. Transient DNA-replication assay revealed that HPV DNA replication is stimulated in a dose-dependent manner by addition of Ini1/hSNF5, and that Ini1/hSNF5 antisense RNA blocks the replication of HPV DNA. Amino-acid substitution at residues that are conserved among E1 proteins prevented the E1-Ini1/hSNF5 interaction and reduced DNA replication of HPV in vivo. Our results indicate that Ini1/hSNF5 is required for the efficient replication of papillomavirus DNA and is therefore needed, either alone or in complex with SWI/SNF complex, for mammalian DNA replication as well. PMID- 10365965 TI - Enzyme dynamics and hydrogen tunnelling in a thermophilic alcohol dehydrogenase. AB - Biological catalysts (enzymes) speed up reactions by many orders of magnitude using fundamental physical processes to increase chemical reactivity. Hydrogen tunnelling has increasingly been found to contribute to enzyme reactions at room temperature. Tunnelling is the phenomenon by which a particle transfers through a reaction barrier as a result of its wave-like property. In reactions involving small molecules, the relative importance of tunnelling increases as the temperature is reduced. We have now investigated whether hydrogen tunnelling occurs at elevated temperatures in a biological system that functions physiologically under such conditions. Using a thermophilic alcohol dehydrogenase (ADH), we find that hydrogen tunnelling makes a significant contribution at 65 degrees C; this is analogous to previous findings with mesophilic ADH at 25 degrees C. Contrary to predictions for tunnelling through a rigid barrier, the tunnelling with the thermophilic ADH decreases at and below room temperature. These findings provide experimental evidence for a role of thermally excited enzyme fluctuations in modulating enzyme-catalysed bond cleavage. PMID- 10365964 TI - Structure and ligand of a histone acetyltransferase bromodomain. AB - Histone acetylation is important in chromatin remodelling and gene activation. Nearly all known histone-acetyltransferase (HAT)-associated transcriptional co activators contain bromodomains, which are approximately 110-amino-acid modules found in many chromatin-associated proteins. Despite the wide occurrence of these bromodomains, their three-dimensional structure and binding partners remain unknown. Here we report the solution structure of the bromodomain of the HAT co activator P/CAF (p300/CBP-associated factor). The structure reveals an unusual left-handed up-and-down four-helix bundle. In addition, we show by a combination of structural and site-directed mutagenesis studies that bromodomains can interact specifically with acetylated lysine, making them the first known protein modules to do so. The nature of the recognition of acetyl-lysine by the P/CAF bromodomain is similar to that of acetyl-CoA by histone acetyltransferase. Thus, the bromodomain is functionally linked to the HAT activity of co-activators in the regulation of gene transcription. PMID- 10365966 TI - Necrotizing fasciitis of the pharynx following adenotonsillectomy. AB - Necrotizing fasciitis is a rare clinical entity in the head and neck region. We report a case of necrotizing fasciitis following adenotonsillectomy in a previously healthy 2-year-old girl. The child presented in a septic state with impending airway compromise. Computed tomography (CT) showed massive soft tissue widening with air in the retropharyngeal, parapharyngeal and retromandibular spaces. Intraoperative exploration showed necrosis of the posterior pharyngeal wall from the skull base to the cricoid, with extension into the parapharyngeal and retropharyngeal spaces. Cultures from the debrided tissues grew two aerobes and three anaerobes. Management involved airway support, surgical debridement, broad spectrum antibiotic coverage and nutritional support. The patient ultimately developed nasopharyngeal and oropharyngeal stenosis requiring tracheostomy and gastrostomy tube placement. This case report highlights an extremely rare complication of adenotonsillectomy. PMID- 10365967 TI - Otoacoustic emission criteria for neonatal hearing screening. AB - Transient evoked otoacoustic emission measures are gaining acceptance as a technique in new-born hearing screening. At present a wide variety of pass-fail screening criteria are used in otoacoustic emission screening programs. In a study of 100 special care neonates and 35 well, full term babies, a number of screening criteria were examined for sensitivity and specificity characteristics when compared to a standard auditory brainstem response protocol. Results indicate that, for normal and special care neonates with a gestational age at test of 38-41 weeks, high sensitivity ( > 80%) could be obtained when a pass-fail criterion involving analysis of emission reproducibility, or emission reproducibility and emission response level, was set. Sensitivity was reduced for special care neonates who fell outside this age range. Specificity was found to be relatively low overall (always < 65%) and may relate to clinical factors in special care neonates not investigated in this study. PMID- 10365968 TI - Results with sphincter pharyngoplasty and pharyngeal flap. AB - OBJECTIVE: To evaluate speech outcomes and complications of sphincter pharyngoplasty and pharyngeal flap performed for management of velopharyngeal insufficiency (VPI). DESIGN: Case series. SETTING: Tertiary care children's hospital. PATIENTS: All patients who underwent pharyngeal flap or sphincter pharyngoplasty from 1990 to 1995. METHODS: Perceptual speech analysis was used to assess severity of VPI, presence of nasal air emissions and quality of nasal resonance (hyper, hypo, or normal). Pre-operative measures of velopharyngeal function were based upon nasendoscopy and videofluoroscopic speech assessment. Recommendations for management were made by the attending surgeon. Complications of hyponasality and obstructive sleep symptoms (OSS) were noted. Patient characteristics were compared using univariate analysis. RESULTS: Sixteen patients underwent sphincter pharyngoplasty and 18 patients underwent superiorly based pharyngeal flap. Patients were similar in terms of lateral pharyngeal wall medial motion and palatal elevation. The groups were also similar with regard to VPI severity, though there was a trend for more severe VPI in patients undergoing sphincter pharyngoplasty than pharyngeal flap (50 vs. 33.3%, respectively). Patients with pharyngoplasty had a higher rate of resolution of VPI than those who had pharyngeal flap (50 vs. 22.2%, respectively), although this was not statistically significant. Post-operative hyponasality and obstructive sleep symptoms were present in both groups. However, only patients who underwent PF and had postoperative OSS had obstructive sleep apnea (OSA). CONCLUSIONS: There were no detectable anatomic differences between treatment groups implying that treatment selection during the study period was not guided by strict anatomic criteria. Sphincter pharyngoplasty may have a higher success rate with a lower risk of OSS. PMID- 10365969 TI - ABR differences before and after dialyses. AB - The aim of the study was the objective evaluation of the hearing organ in children. Seven children aged 6-17 with end stage renal failure treated by haemodialysis were examined. Four- to five-hour dialyses were performed 2-3 times a week. Pure-tone audiometry was normal in five children. The reactions of brain stem responses (latency and interpeak latency) were analysed before and after haemodialyses. We conclude that the ABR differences were connected with biochemical parameters of uraemia. The present study is a preliminary one and will be continued in the future. PMID- 10365970 TI - Temporal bone histopathologic findings in a case of interstitial deletion of the long arm of chromosome 2 [del(2) (q31q33)]. AB - The temporal bones of a 24-day-old female neonate with an interstitial deletion of the long arm of chromosome 2 [del(2)(q31q33)] were studied histopathologically, focusing mainly on the inner ear. This is, to our knowledge, the first report of a temporal bone study in a case of this chromosomal aberration. The abnormalities included a shortened cochlea with underdeveloped modiolus in both ears. Total absence of the spiral ganglion cells and the cochlear nerve bundle, accompanied by obliteration of the fundus of the internal auditory meatus by a bony plate, and dislocation of some geniculate ganglion cells to the internal auditory meatus were also observed in the right ear. The absence of the spiral ganglion cells is considered to be the result of some complication in the early fetal life, such as dysgenesis or early degeneration of the neuronal cells. The organ of Corti in the right ear was well-developed and preserved in the middle turn, suggesting that the differentiation of the cochlear sensory epithelium in humans is not dependent upon the innervation, at least on the gross level. PMID- 10365971 TI - Techniques for improving ear definition in microtia reconstruction. AB - Surgeons involved in microtia repair recognize the difficulty in creating a natural appearing ear. One key to successful reconstruction is to provide sufficient relief between the helix, scaphoid fossa and antihelix to create the illusion of thin skin overlying thin cartilage. Problems such as thick skin, hair bearing skin and poor-quality cartilage serve to frustrate the surgeons attempt to achieve the desired result. Surgical techniques to improve cartilaginous framework definition in microtia repair are discussed. PMID- 10365972 TI - Mucoepidermoid carcinoma arising in the accessory parotid gland. AB - A rare case of a 9-year-old female with mucoepidermoid carcinoma arising in the accessory parotid gland is reported. She had complained of a painless and round mass of the left cheek for a duration of 14 months. Sialography, ultrasonography, CT scan and MRI were performed preoperatively. Sialography revealed a small duct separating from the Stensen's duct. CT and MRI showed that the tumor with smooth outline was lying on the masseter muscle and detached from the main parotid gland. The preoperative diagnosis was an accessory parotid gland tumor. The tumor was removed without facial nerve injury via standard parotidectomy incision. The tumor was composed of mucous and epidermoid cells. The pathological diagnosis was low-grade mucoepidermoid carcinoma. PMID- 10365973 TI - Central deafness in a young child with Moyamoya disease: paternal linkage in a Caucasian family: two case reports and a review of the literature. AB - A case of 'central deafness' is presented in a 3-year-old male Caucasian child with Moyamoya disease (MMD); a rare, progressive and occlusive cerebrovascular disorder predominantly affecting the carotid artery system. Documentation of normal peripheral auditory function and brainstem pathway integrity is provided by acoustic admittance, otoacoustic emission and brainstem auditory evoked potential measurements. The lack of behavioral response to sound, and absent middle and long latency auditory evoked potentials suggest thalamo-cortical dysfunction. Magnetic resonance imaging showed diffuse ischemic damage in subcortical white matter including areas of the temporal lobes. In addition, there were multiple and focal cortical infarctions in both cerebral hemispheres, focused primarily in the frontal, parietal and temporal areas. Taken together, these structural and functional abnormalities in addition to severely delayed speech and language development are consistent with the diagnosis of central deafness and suggest a disconnection between higher brainstem and cortical auditory areas. The child's father also has MMD, but was diagnosed only recently. The presence of paternal linkage is informative since it rules out x-linked recessive and maternal inheritance. To our knowledge, this represents the first documented case of paternal linkage in MMD with central deafness in a Caucasian child with no apparent Japanese ancestry. Herein, we focus on central auditory dysfunction and consider how lesion-induced changes have contributed to a deficit in basic auditory responsiveness, including a severe disturbance in receptive and expressive auditory-based speech and language skills. PMID- 10365974 TI - Dermoid cyst of the eustachian tube. AB - We report a case of dermoid cyst of the Eustachian tube in a 2 1/2 -year-old-girl with CT and MRI imaging. This is the 12th described case of such a pathology. Most of the reviewed previous 11 cases affected females on the left side. The surgical approach and the contribution of CT and MRI are discussed. PMID- 10365975 TI - Langerhans' cells histiocytosis. AB - Langerhans cell histiocytosis (LCH) is a rare disorder of unknown cause, characterized by the proliferation of histiocytic cells in various tissues and organs. The role of the otolaryngologist is important in the early and accurate evaluation, staging and diagnosis of LCH, because it may mimic more common diseases such as otitis externa and acute mastoiditis. We discuss a case report of bilateral mastoid involvement in a child with a history of otalgia unresponsive to medical therapy. PMID- 10365976 TI - The nature and significance of nutritional adaptation. AB - Possible adaptations to a low protein intake are: a decrease in the obligatory nitrogen loss, which would be too small to detect in short-term studies, but would be significant over a longer term; an increase in the efficiency of protein utilization, which has been demonstrated in depleted subjects; and a decrease in lean body mass, mainly at the expense of muscle. However, we do not know the extent to which this last mechanism may really be an adaptation without significant functional loss. In the case of energy there is controversy about the extent to which the gross efficiency of muscular work can be improved. One mechanism might be an alteration in the distribution of fibre types, with a shift from fast to slow fibres. A possible way of reducing the cost of both muscular work and basal metabolism would be a reduction in the mitochondrial proton leak. Both these mechanisms are at least partially under the control of the thyroid gland, which therefore may play an important role in economizing energy expenditure. PMID- 10365977 TI - Molecular and cellular adaptations to carbohydrate and fat intake. AB - Adaptation to carbohydrate and fat intake involves changes in a number of biochemical parameters at the cellular level. A change in the concentration of fat or carbohydrate in the blood acts directly to influence metabolic pathways by altering the flux of intermediates into cells. This in turn alters the concentration of hormones and other signaling molecules and changes the rate of expression of genes coding for key regulatory proteins or enzymes in metabolic pathways. These effects occur at different rates and in a tissue-specific manner in response to diet. A key metabolic adaptation involves changes in the level of expression of genes coding for proteins of critical importance in energy metabolism; this is largely due to an altered rate of transcription of selected genes under the control of hormones and/or carbohydrate and lipid. The mediators of this effect are transcription factors, that is, nuclear proteins which integrate the effects of hormones and substrates with the transcription process by binding to response elements in the promoters of regulated genes and interacting with the transcription machinery at the TATA box, thereby altering the activity of RNA polymerase II. In this review we will outline the hierarchy of cellular adaptations to diet and will emphasize the latest concepts of gene regulation in response to metabolites and hormones. In particular, we will review the role of the various transcription factors involved in the regulated expression of the gene for the cytosolic form of phosphoenolpyruvate carboxykinase (GTP) (EC 4.1.132), a key gluconeogenic enzyme. PMID- 10365978 TI - Adaptation to low energy intakes: the responses and limits to low intakes in infants, children and adults. AB - Reduction in energy intake below the acceptable level of requirement for an individual results in a series of physiological and behavioural responses, which are considered as an adaptation to the low energy intake. This ability of the human body to adapt to a lowering of the energy intake is without doubt beneficial to the survival of the individual. However, what is more controversial is the view held by some that the body can metabolically adapt in a beneficial manner to a lowered intake and consequently that the requirements for energy are variable given the same body size and composition and physical activity levels. Much of this confusion is the result of considerable evidence from studies conducted in well-nourished adults who, for experimental or other reasons, have lowered their intakes and consequently demonstrated an apparently enhanced metabolic efficiency resulting from changes in metabolic rates which are disproportionate to the changes in body weight. Similar increases in metabolic efficiency are not readily seen in individuals who on long-term marginal intakes, probably from childhood, have developed into short-statured, low-body-weight adults with a different body composition. It would thus appear that the generally used indicator of metabolic efficiency in humans, that is a reduced oxygen consumption per unit fat free mass, is fraught with problems since it does not account for variations in contributions from sub-compartments of the fat free mass which include those with high metabolism at rest such as brain and viscera and those with low metabolism at rest such as muscle mass. Metabolic rate per unit fat free mass thus, does not reflect true variations in metabolic efficiency and is due largely to variations in body composition. This finding combined with the evidence that behavioural adaptation in habitual physical activity patterns which occurs on energy restriction is not necessarily beneficial to the individual raises doubts about the role of adaptation to low intakes in determining one's requirement for energy. The evidence is overwhelming that both in children and adults, changes in body size and composition as well as in levels of habitual physical activity may be the most important consequences of a lowered energy intake and cannot be assumed to be a part of a beneficial adaptation that influences energy requirements. PMID- 10365979 TI - Adaptation of protein metabolism in relation to limits to high dietary protein intake. AB - Studies of the effects of dietary protein level on human metabolism have usually concentrated on the effects of protein deprivation and on establishing a minimum dietary requirement. By contrast, less is known about the effects of very high protein diets, although general levels of protein intake in the developed world are increasing, and high protein diets have been advocated for maintaining or increasing muscle mass in certain groups of the population. This article, therefore, examines the response of protein metabolism to high dietary protein, studied in adults by nitrogen balance and isotopic tracer techniques, and concentrating on the evidence for increased lean body mass. It is concluded that high protein feeding initially results in protein retention, with greater cycling of body protein in response to meals, but that neither N-balance nor isotopic tracer methods possess sufficient sensitivity to detect whether a long term increase in functional lean tissue ensues. Improved methods of body composition measurement will be needed to establish this. Moreover, the absence of strong evidence that high protein diets confer any advantage in terms of strength or health must be weighed against potentially injurious consequences. PMID- 10365981 TI - De novo lipogenesis in humans: metabolic and regulatory aspects. AB - The enzymatic pathway for converting dietary carbohydrate (CHO) into fat, or de novo lipogenesis (DNL), is present in humans, whereas the capacity to convert fats into CHO does not exist. Here, the quantitative importance of DNL in humans is reviewed, focusing on the response to increased intake of dietary CHO. Eucaloric replacement of dietary fat by CHO does not induce hepatic DNL to any substantial degree. Similarly, addition of CHO to a mixed diet does not increase hepatic DNL to quantitatively important levels, as long as CHO energy intake remains less than total energy expenditure (TEE). Instead, dietary CHO replaces fat in the whole-body fuel mixture, even in the post-absorptive state. Body fat is thereby accrued, but the pathway of DNL is not traversed; instead, a coordinated set of metabolic adaptations, including resistance of hepatic glucose production to suppression by insulin, occurs that allows CHO oxidation to increase and match CHO intake. Only when CHO energy intake exceeds TEE does DNL in liver or adipose tissue contribute significantly to the whole-body energy economy. It is concluded that DNL is not the pathway of first resort for added dietary CHO, in humans. Under most dietary conditions, the two major macronutrient energy sources (CHO and fat) are therefore not interconvertible currencies; CHO and fat have independent, though interacting, economies and independent regulation. The metabolic mechanisms and physiologic implications of the functional block between CHO and fat in humans are discussed, but require further investigation. PMID- 10365980 TI - Limits of adaptation to high dietary protein intakes. AB - Ingested protein is made available to the body following digestion and absorption as amino acids and contributes to the body's demand for amino acids for protein synthesis and other metabolic pathways. As the pattern of amino acids required for metabolism is substantially different from that ingested, extensive metabolic interchange serves to improve the match. As a matter of course oxidation of amino acids contributes to satisfying the energy needs of the body. Amino acids in excess of immediate requirements follow degradative pathways and if the capacity of these pathways is exceeded adverse consequences ensue. In pathological states, such as inborn errors of metabolism, there is an obvious constraint on metabolic flow with serious sequelae. Pathways may be constrained to a lesser extent due to genetic polymorphisms, metabolic programming, limitation of cofactors or lack of associated substrates. Any of these can result in metabolic derangements, which do not manifest as overt disease, but limit normal function. There is the need to determine the dose response to increases in dietary protein and amino acid availability, using critical metabolic intermediates as outcome indices in order to clarify the upper limit of intake with which the body can cope under a range of physiological and pathological states. PMID- 10365982 TI - Essential fatty acids as determinants of lipid requirements in infants, children and adults. AB - Essential fatty acids (EFA) are the indispensable component of the lipid supply beyond the provision of energy as a fuel for oxidation. They serve as dietary precursors for the formation of prostanoids and other eicosanoids thus are of great significance in health and modulation of disease conditions. Eicosanoids are powerful autocrine and paracrine regulators of cell and tissue functions: thrombocyte aggregation, inflammatory reactions and leukocyte functions, vasoconstriction and vasodilatation, blood pressure, bronchial constriction, and uterine contraction. Recent attention has focused on the effect of n-3 and n-6 long chain EFAs in normal fetal development. Results from human infant studies suggest that n-3 fatty acids are needed for optimal development of visual and brain function. Human milk is the best and only time proven source of fat and dietary essential fatty acids for infant feeding. International recommendations for n-3 and n-6 EFA dietary intake are reviewed and suggested intakes for long chain EFAs are provided. PMID- 10365983 TI - Response to and range of acceptable fat intakes in infants and children. AB - The maximum and minimum fat intakes that are physiologically tolerable by healthy infants and young children are not well defined. The maximum tolerable fat intake appears to be limited by the minimum requirements of protein, carbohydrates and micronutrients. It is unresolved whether or not there is a minimum metabolic requirement of dietary fat, beyond the requirements of essential fatty acids and lipid soluble vitamins and the effects on energy density and an adequate total energy intake. The first postnatal months of the human infant are characterized by a rapid weight gain and extensive fat deposition. Body fat deposition equals 25% of the total energy intake during the first 2 and 16% during the third and fourth months, respectively. The provision of dietary lipids in amounts at least matching the needs for tissue storage and fat oxidation appears to be of advantage for energy balance and physiological growth. Some reports have associated low fat diets with less than 30% of energy as fat with adverse effects on child growth beyond infancy, but it remains unresolved whether these effects were caused by an associated effect on dietary energy density and total intake of energy and other nutrients. Closely supervised, healthy infants from an affluent population grew normally with a diet providing about 30% of the energy as fat from the seventh month of life onwards, but it is not known to what extent adaptive mechanisms such as a reduction of physical activity may have been required. It is not known whether infants and young children stressed by frequent occurrence of diarrhea and infections may adapt to a 30% fat diet as well or not. No evidence is available for a health benefit of a low total fat intake in infancy. In view of the limited available information, further research is required to define optimal fat intakes in early childhood since this question is of major importance for child health. PMID- 10365984 TI - Response to and range of acceptable fat intake in adults. AB - Cellular energy metabolism depends on two main energy substrates: glucose and fatty acids. The major determinants of the fuel mix oxidized are glucose availability and insulin secretion that both promote glucose oxidation. Fatty acid oxidation occurs mainly when glucose availability is reduced, for instance during the postabsorptive period, or when energy expenditure is increased, for instance during exercise of long duration. When eucaloric diets with high carbohydrate and low fat content are ingested, de novo lipogenesis is stimulated in adults, but the rate of conversion of glucose to fatty acids is low, which means that carbohydrate intake does not have much influence on fat requirements. The lower limit of fat intake depends on three factors: the fat requirement to meet energy needs, the need for essential fatty acids, and the amount of fat in the diet that is necessary to absorb fat-soluble vitamins. The lower limit of fat intake to meet the energy needs of adults is assumed to be between 10 and 15% of dietary energy, provided that enough carbohydrates are available. For adults, the requirement for essential fatty acids is in the range of 3-5% of dietary energy for linoleic acid, and 0.5-1.0% of dietary energy for linolenic acid. Fat energy should not be below 10% of total energy intake in order to ensure an unrestricted absorption of fat-soluble vitamins, particularly vitamins A and E. The recommendations on upper limits of fat intake for adults must take into account the degree of physical activity. International recommendations indicate that active individuals in energy balance may consume up to 35% of their total energy intake as dietary fat, whereas sedentary individuals should not consume more than 30% of their energy from fat. Saturated fatty acids should not exceed 10% of the energy intake. PMID- 10365985 TI - Carbohydrate as nutrient in the infant and child: range of acceptable intake. AB - Carbohydrates are the major source of energy for humans. Following their digestion, almost all ingested carbohydrates are converted to glucose. Glucose is the primary oxidative fuel for the brain. Although few studies have been done in infants and children to define the upper and lower limits of carbohydrate intake, such information may be derived from the published data on glucose metabolism in vivo. The upper and lower limits are determined by the need to provide for total energy expenditure, need for other essential nutrients such as protein and fats, requirements of the glucose dependent tissues such as the brain, and the need to minimize the protein cost of gluconeogenesis and thus irreversible loss of nitrogen. With these considerations, the upper and lower boundaries of carbohydrate intake in relation to age are described. PMID- 10365986 TI - Carbohydrate as a nutrient in adults: range of acceptable intakes. AB - This review considers the acute and chronic effects of different levels of carbohydrate (CHO) intakes. The type of CHO consumed, especially glucose vs fructose, affects the glycaemic, insulinaemic and thermogenic responses. In addition, other aspects of food (type of starch, method of processing or cooking, presence of other nutrients) affects the glycaemic response (glycaemic index). In general, the greatest benefit to health is derived from consuming foods with a low glycaemic index and a high non-starch polysaccharide (fibre) content. Healthy, moderately active adults require at least 200g CHO per day to sustain normal brain metabolism and muscle function. Moreover, the CHO content should represent at least 50% of energy intake. Higher intakes of CHO can have deleterious effects on blood lipids (especially plasma triacylglycerol) in middle aged and elderly subjects, and are really only appropriate for subjects with a high level of physical activity who need to maintain muscle glycogen content. Meals with a high carbohydrate content can lead to problems of postprandial hypotension in the elderly, and impaired exercise capacity in patients with angina. PMID- 10365987 TI - Comments on metabolic needs for glucose and the role of gluconeogenesis. AB - The metabolism of carbohydrates is largely determined by their chemical properties. Glucose may have been selected, over the other aldohexoses, because of its low propensity for glycation of proteins. That carbohydrate is stored in polymeric form (glycogen) is dictated by osmotic pressure considerations. That stored fat is about eight times more calorically dense than glycogen, when attendant water is factored in, accounts for the predominance of fat as a storage form of calories and, also, for the fact that ingested carbohydrate is oxidized promptly (that is, fuel of the fed state) rather than being extensively stored. That stored glycogen is accompanied by so much water accounts for the fact that the brain only has very small glycogen stores. Carbohydrate has two important advantages, over fat, as a metabolic fuel; it is the only fuel that can produce ATP in the absence of oxygen, and more ATP is produced per O2 consumed when glucose is oxidized, compared with when fat is oxidized. These advantages probably determine the preference of many cell types for carbohydrate. In addition to its use as a metabolic fuel, glucose plays other important roles such as provision of NADPH via the pentose phosphate pathway, and as a source material for the synthesis of other key carbohydrates, for example, ribose and deoxyribose for nucleic acid synthesis and substrates for the synthesis of glycoproteins, glycolipids and glycosaminoglycans. It can also play a key role in anaplerosis. Although it is widely acknowledged that gluconeogenesis plays a crucial role in starvation it is now apparent that prandial gluconeogenesis occurs, both in the metabolic disposal of dietary amino acids and in the synthesis of glycogen by the indirect pathway. Although there is, strictly speaking, no dietary requirement for carbohydrate it is evident that glucose is a universal fuel for probably all cells in the body and carbohydrate is the cheapest source of calories and the major source of dietary fibre. These observations, together with the fact that glucose is the preferred metabolic fuel for the brain, permit us to recommend appreciable quantities of carbohydrate in all prudent diets. PMID- 10365988 TI - Physical exercise as a modulator of adaptation to low and high carbohydrate and low and high fat intakes. AB - Quantification of the metabolic response aids in ascertaining the nature and extent of the energy requirements imposed by exercise. During high intensity exercise, virtually all of the energy is supplied by the net oxidation of glycogen while fat oxidation plays a more prominent role during lower intensity exercise. Therefore, the lower limit of carbohydrate required above resting needs is equal to the portion of the total energy cost derived from carbohydrate sources. There is no upper limit of additional carbohydrate intake that could be eaten to satisfy the extra caloric requirement since carbohydrate intake will restore any endogenous energy stores that were used during exercise, regardless of the intensity of exercise. The recommendation of a high carbohydrate intake to provide caloric balance in exercising individuals is supported by the observation that exercise performance at high intensity is improved by a high carbohydrate diet, and exercise performance at low intensity is relatively insensitive to the source of the caloric intake. Limited dietary studies are consistent with predictions based on the metabolic response. At exercise intensities below 65% VO2 max, the percent fat and carbohydrate in the diet makes little difference on exercise performance, provided adequate time is allowed to adapt to a high-fat diet. On the other hand, exercise ability during high-intensity exercise is significantly limited by a high-fat diet. A consideration of importance beyond the aspect of energy balance is the anabolic effect of insulin on muscle protein synthesis after exercise. Provision of carbohydrate after exercise is likely to stimulate muscle protein synthesis to a greater extent than a corresponding amount of fat. Dietary fats may offer practical advantages to the athlete but if fats are consumed at the expense of carbohydrate intake, many established benefits of high carbohydrate intake in terms of performance may be sacrified. PMID- 10365989 TI - Adaptation of maternal lipid flux to pregnancy: research needs. AB - Adaptations of maternal lipid metabolism during pregnancy are directed toward both the needs of the fetus for lipid substrates and maternal requirements for lipid stores serving as energy reserves for lactation. The mechanisms are poorly understood, but must be elucidated before new dietary recommendations can be made about dietary supplementation with long-chain polyunsaturated fatty acids (LC PUFA). Problems of high priority for research are: (1) Fetal requirements for specific fatty acids, including essential fatty acids (EFA) and long-chain polyunsaturated fatty acids (LC-PUFA); (2) The mechanisms for transfer of fatty acids across the placenta; (3) The role of very low density lipoproteins (VLDL) in transfer of EFA and LC-PUFA from the maternal liver to the placenta; (4) Adjustments of fatty acid metabolism in the maternal liver during pregnancy; and (5) The effect of dietary LC-PUFA on maternal fatty acid metabolism during pregnancy. PMID- 10365990 TI - Pregnancy and lactation in relation to range of acceptable carbohydrate and fat intake. AB - The additional energy requirements of pregnancy are needed for increases in maternal (breast, uterus and adipose) and feto-placental tissue accrued during pregnancy as well as the additional running cost of pregnancy for example increased cardiac output. Based on prospective longitudinal studies, the additional energy requirements of pregnancy range from > 500 MJ in Swedish women to net savings of approximately 50 MJ in women in The Gambia with their usual nutritional intake. In addition to the wide variation in estimated energy expenditure among various ethnic populations, there is as much as a 10-20 fold range in the total energy cost of pregnancy and lactation within relatively homogenous populations. The estimates of energy intake in these studies, however, are generally less than the estimates of total energy expenditure. The discrepancy between energy intake and energy expenditure during pregnancy is most probably due to several factors including decreased maternal activity, unreliable reporting of energy intake and possibly increased metabolic efficiency of basal metabolic rate, thermic effect of foods and physical activity. Based on recent studies, variations in maternal pregravid glucose insulin sensitivity may account for part of the observed variability associated with maternal metabolic adaptations during pregnancy. Decreases in insulin sensitivity have a significant inverse correlation with accretion of adipose tissue in early pregnancy. Likewise, there is a significant inverse correlation between decreases in basal oxygen consumption with increases in endogenous glucose production. The mechanism for these changes remain speculative. Additionally, although serum leptin concentrations increase 66% in early pregnancy and are correlated with maternal fat mass and basal energy expenditure, the increases in serum leptin occur prior to any significant increases in body fat or basal metabolic rate suggesting that pregnancy represents another leptin resistant state. Based on these data, specific recommendations for acceptable carbohydrate and fat intake during pregnancy and lactation are not possible for every woman at this time. Additional prospective studies, evaluating long-term maternal and neonatal outcome are needed before more meaningful nutritional recommendations can be proposed. PMID- 10365991 TI - Sepsis as a modulator of adaptation to low and high carbohydrate and low and high fat intakes. AB - Catabolism of lean body mass (particularly muscle) occurs in sepsis and other forms of critical illness despite apparently adequate nutritional support. The determination of the optimal balance of carbohydrate and fat intake in this circumstance should be based on the resulting effect on the maintenance of lean body mass, and the nature and extent of any side effects. The general stress response involves a disruption in normal glucoregulation, in that hepatic glucose production is accelerated and the normal blood glucose lowering action of insulin is diminished. Nonetheless, the capacity to oxidize glucose once inside the cells is not impaired. Lipolysis, or the breakdown of peripheral triglycerides to free fatty acids (FFA) and glycerol, is accelerated in critical illness, to a greater extent than fat oxidation. Provision of exogenous fat maintains fat stores, but has minimal effect on the direct oxidation of plasma FFA. From the results of oxidation studies, it seems that about 5 mg kg x min of glucose can be readily oxidized, and the balance of energy will be supplied by the oxidation of fat, either endogenous or exogenous. However, an additional consideration in determining the optimal caloric substrate is that insulin is a potent anabolic hormone and stimulates muscle protein synthesis. Consequently, provision of exogenous insulin enhances retention of muscle. This procedure dictates that almost all non-protein calories be provided as carbohydrate to avoid hypoglycemia. Preliminary studies suggest this may be the optimal approach in critically ill patients. Glucose and fatty acids are the major energy substrates in the body. The oxidative metabolism of these substrates provides the ATP needed for physiological function, including protein synthesis. Over the past 20 y, development of new techniques in nutritional support have made it possible to provide large amounts of carbohydrate and fat to critically-ill patients, along with protein or amino acids. However, despite providing such patients with what should be more than adequate caloric and protein intake, critically ill patients lose lean body mass (Streat et al, 1987), largely because of persistent muscle catabolism (Sakurai et al, 1995). The general relation between energy substrate metabolism and maintenance of lean body mass has been recognized for many years (Calloway & Spector, 1954), so it is important to examine the alterations in energy substrate metabolism that occur in response to critical illness that may play a role in causing the persistent catabolism of muscle protein. PMID- 10365992 TI - Limitations of nutrient intake. The effect of stressors: trauma, sepsis and multiple organ failure. AB - The response to injury includes a diminution in appetite, a decrease in nutrient intake, an acute mobilisation of endogenous energy stores (glucose and fat), but an impaired ability to use them. Lean tissue is broken down to its constituent amino acids, which provide precursors for the synthesis of glucose in the liver (gluconeogenesis). Glucose is used as a source of energy by the brain and red blood cells, as well as by wound tissue. After a discrete injury normal function is normally resumed with a reduced body mass. In very severe injury or sepsis, in those who are physiologically or immunologically impaired or those with a genetic predisposition to the condition, organ failure may develop due to an apparent ongoing inflammatory process. The origin of this process is not always apparent, but loss of integrity of the gastrointestinal tract has been suggested. Apparently adequate nutritional support in the presence of a severe inflammatory stimulus only attenuates the gluconeogenic process, and the breakdown of lean tissue continues. Supply of protein (amino acids) stimulates protein synthesis, but it also stimulates breakdown. Nutrient intake via the enteral route may be limited by gastrointestinal symptoms and via the parenteral route by fluid overload, although this can be circumvented by fluid removal by haemofiltration. It is probable that, if nutritional support in severe trauma/sepsis/multiple organ failure is to be effective, satisfactory pharmacological methods of controlling metabolism will have to be found. PMID- 10365993 TI - High and low carbohydrate and fat intakes: limits imposed by appetite and palatability and their implications for energy balance. AB - This report examines several issues concerning the effects of dietary fats and carbohydrates (CHOs) on body weight and the limits set on the intake of these nutrients by factors influencing appetite control: (i) the physiological relationship between feeding behaviour (FB) and body weight; (ii) the distribution of nutrients in Western foods and the implications this may have for FB; (iii) the contribution of nutrients in the diet, to total EI under both extreme and typical Western conditions; (iv) the known effects that fats, CHOs and dietary energy density (ED) exert on appetite and energy balance (EB); (v) the potential role of sensory factors in promoting or limiting fat, CHO and energy intakes (EI) in modern human populations Population studies and large surveys have identified individuals with wide ranges of fat and CHO intakes. Intakes of fat can vary from an average of 180 g/day to 25 g/day in a representative sample. But on individual days fat intake can rise to well over 200 g according to a selection of high fat foods. In a single meal, people can consume an amount of fat greater than the population daily average. From this analysis it can be deduced that the appetite control mechanism will permit the consumption of large amounts of fat (if an abundance of high fat foods exist in the food supply). Except for specific physiological circumstances (e.g. endurance explorers) where there is an urgent need for EIs, in the face of decreasing body weight, it is unlikely that the body will generate a specific drive for fat. Because CHO foods have a lower ED than fat foods (on average) and because of their greater satiating capacity, the free intake of high CHO foods is likely to be self-limiting (at lower EIs than those generated by fatty foods). This does not mean that excess EIs are impossible when people feed ad libitum on high CHO diets. PMID- 10365994 TI - Intake of fat and carbohydrate: role of energy density. AB - In this review, we consider two hypotheses which could explain why high-fat foods are overeaten. The first hypothesis is that fat is overeaten because it affects satiety and satiation less than carbohydrate. In several studies which have evaluated the effects of fat on satiety and satiation, fat differed little from carbohydrate when both the palatability and energy density of the test foods were matched. Therefore it is unlikely that the effects of fat on satiety or satiation provide the primary explanation for why it is overeaten. The second hypothesis is that the high energy density of fat facilitates its overconsumption. Support for this view comes from recent studies in which energy density significantly influenced intake when both the macronutrient content and palatability of the test foods were matched. For example, when individuals were fed diets varying in energy density and could eat as much food as they liked, they ate the same amount of food (by weight) so energy intake varied directly with energy density. Furthermore, when participants consumed foods of low energy density, they felt satisfied, despite reductions in energy intake. These findings show that energy density is a key determinant of energy intake in that cognitive, behavioral, and sensory cues related to the volume or weight of food consumed can interact with or override physiological cues associated with food intake. PMID- 10365995 TI - Report of the IDECG Working Group on lower limits of energy and protein and upper limits of protein intakes. International Dietary Energy Consultative Group. PMID- 10365996 TI - Report of the IDECG Working Group on lower and upper limits of carbohydrate and fat intake. International Dietary Energy Consultative Group. PMID- 10365997 TI - Report of the IDECG Working Group on the modulating effect of stressors on the upper and lower limits of lipid and carbohydrate intake. International Dietary Energy Consultative Group. AB - The recommended caloric intake should be increased by 2870kJ (670Kcal)/d in the exclusively breast-feeding woman. Of this increase, 10% should be ingested as protein, up to 50% as lipid and no less than 40% as carbohydrate. If weight loss is desired lipid intake can be reduced but carbohydrate intake should be maintained. Of the total energy increment 3% should be supplied by essential fatty acids (linoleic and alpha-linolenic acids) and 0.5% by n-3 fatty acids. The issue of dietary supplementation with long-chain polyunsaturated fatty acids remains for future research. PMID- 10365998 TI - A new method for magnetoencephalography: a three-dimensional magnetometer-spatial filter system. AB - A novel three-dimensional magnetometer-spatial filter system was developed to study human brain activity with high spatiotemporal resolution. The system combines the high temporal resolution of magnetoencephalography with the high spatial resolution achieved by using three-dimensional magnetometers and spatial filters to measure the direction and intensity of magnetic fields generated during brain activity. Simulation and phantom studies indicate that the system is capable of mapping current sources of magnetic fields with a spatial resolution comparable to that of any other brain functional imaging technique while maintaining millisecond temporal resolution. Application of this system to the human brain resolved magnetoencephalographic responses evoked by motion stimuli on a millisecond scale into responses occurring in visual cortical areas V1, V2/3 and V5. It also revealed signals related to contextual modulation in V1 and V2/3. This system provides a new way of studying the dynamics of human brain function. PMID- 10365999 TI - Common dynamics in temporal lobe seizures and absence seizures. AB - Similarities among the clinical features of complex partial temporal lobe seizures and absence (petit mal) seizures suggest shared underlying mechanisms, but dissimilar electrographic features of the two seizure types have cast doubt on common neuronal substrates. However, visual inspection and traditional approaches to quantitative analysis of the electroencephalogram and electrocorticogram, such as Fourier analysis, may not be appropriate to identify and characterize the highly non-linear mechanisms likely to underlie ictal events. We previously introduced a technique, non-linear autoregressive analysis, that is designed to identify non-linear dynamics in the electroencephalogram [Schiff N. D. et al. (1991) Society of Neuroscience 21st Annual Meeting, 638.6; Schiff N. D. et al. (1995) Biol. Cybern. 72, 519-526, 527-533]. The non-linear autoregressive analysis technique is aimed at describing seizure discharges as a disturbance of synchrony at the level of neuronal circuits. In absence seizures, we showed that non-linear autoregressive analysis revealed a consistent "fingerprint" of these non-linearities in 3/s discharges within and across patients. Here, we investigate the possibility that non-linear autoregressive modeling of seizure records from patients with temporal lobe epilepsy might reveal common circuit mechanisms when compared with the non-linear autoregressive analysis fingerprint of absence seizures. Electrocorticographic records of seizure activity were obtained in four patients who had received subdural grids or strips implanted in preparation for epilepsy surgery. Decomposition of the multichannel data recorded from these patients by principal component analysis revealed that at least three to five independent "generators" were required to model the data from each patient. Non-linear autoregressive analysis of these extracted generators revealed non-linear dynamics in two patients. In both patients, the temporal aspects of these non-linearities were similar to the characteristic non-linearities identified in the non-linear autoregressive analysis fingerprint of absence seizures. In particular, both patients showed a non-linear interaction of signals 90 ms in the past with signals 150 ms in the past. This was the most prominent interaction seen in all patients with absence seizures (typical and atypical). These results suggest that seizures from some patients with temporal lobe epilepsy may share common underlying circuit mechanisms with those of absence seizures. Physiological interpretations of these results are considered and proposed mechanisms are placed into the context of the alterations of consciousness seen in both epilepsies. PMID- 10366000 TI - Neuronal loss in functional zones of the cerebellum of chronic alcoholics with and without Wernicke's encephalopathy. AB - This study examines the effect of chronic alcohol consumption on the human cerebellum using operational criteria for case selection [Caine D. et al. (1997) J. Neurol. Neurosurg. Psychiat. 62, 51-60] and unbiased stereological techniques. We describe, for the first time, structural changes in different functional zones of the cerebellum of chronic alcoholics and correlate these changes with specific clinical symptoms. No consistent changes in the number of neurons or the structural volume for any cerebellar region were observed in the chronic alcoholics without the clinical signs of Wernicke's encephalopathy. In all cerebellar measures, these chronic alcoholics did not differ significantly from the non-alcoholic controls, suggesting that chronic alcohol consumption per se does not necessarily damage human cerebellar tissue. However, several cerebellar changes were noted in the thiamine-deficient alcoholics studied. There was a significant decrease in Purkinje cell density (reduced on average by 43%) and molecular layer volume (reduced by 32%) in the cerebellar vermis in all thiamine deficient chronic alcoholics. A decrease in cell density and atrophy of the molecular layer, where the dendritic trees of the Purkinje cells are found, without significant cell loss suggests loss of cellular dendritic structure and volume. These thiamine-deficient alcoholics also had a significant decrease (36% loss) in the estimated Purkinje cell number of the flocculi, disrupting vestibulocerebellar pathways. These results indicate that cerebellar Purkinje cells are selectively vulnerable to thiamine deficiency. There is evidence that this damage contributes significantly to the clinical signs of Wernicke's encephalopathy. There was a 36% loss of Purkinje cells in the lateral lobe in alcoholics with mental state signs and 42% atrophy of vermal white matter in ataxic alcoholics. The finding of a 57% loss of Purkinje cells and a 43% atrophy of the molecular layer of the vermis in alcoholics with cerebellar dysfunction supports previous findings highlighting the importance of spinocerebellar pathways to these symptoms. PMID- 10366001 TI - Altered spatial patterns of functional thalamocortical connections in the barrel cortex after neonatal infraorbital nerve cut revealed by optical recording. AB - In rodents, the somatosensory cortex has a cell aggregation cluster termed the barrel, reflecting a whisker vibrissa, and this barrel formation is disrupted by infraorbital nerve cut at birth. In the present study, we prepared thalamocortical slice preparations from rats that received infraorbital nerve cut either at birth or at postnatal day (P) 7 and those from normal rats, recorded the optical response reflecting neural excitation in the somatosensory cortex with a voltage-sensitive dye (RH482) and compared the optical responses from lesioned rats with those from normal rats. In normal rats at P10, the optical response elicited electrically by thalamic stimulation propagated to the cortex, and then several patchy clusters appeared in layer IV. The size and location of these patchy responses precisely matched either barrels identified by cytochrome oxidase staining or terminal arbors of thalamocortial axons stained with biotinylated dextran amine. In contrast, at P10 in P0-lesioned rats, clusters having a wider horizontal width but smaller amplitude than those seen in normal rats appeared in layer IV. Correspondingly, neither cytochrome oxidase staining nor biotinylated dextran amine labeling of thalamocortical axons showed any barrel-like clusters or glomerular axon terminals. Likewise, at P5-P6, the tangential width of clusters in layer IV were larger than that in normal rats. At P10 in P7-lesioned rats, small cluster-matched barrels were seen in the optical response as well as in normal rats. These results suggest that P0 infraorbital nerve cut interrupted segregation of functional synapses into the barrels and retarded the maturation of thalamocortical transmission. PMID- 10366002 TI - GABA release from suprachiasmatic nucleus terminals is necessary for the light induced inhibition of nocturnal melatonin release in the rat. AB - The daily rhythm of melatonin production in the mammalian pineal is driven by the endogenous circadian pacemaker in the suprachiasmatic nuclei. The major release period of melatonin is closely linked to the dark phase of the 24-h day/night cycle. Environmental light will affect melatonin release in two ways: (i) it entrains the rhythm of the circadian oscillator; and (ii) it causes an acute suppression of nocturnal melatonin release. These two effects of light are both mediated by the suprachiasmatic nucleus and enable the pineal gland to convey information about day length to the reproductive system through changes in melatonin levels. Glutamate is currently believed to be the major transmitter in the retinal ganglion cell fibers reaching the suprachiasmatic nucleus. At present no information is available, however, about the transmitter(s) implicated in the further propagation, i.e. from the suprachiasmatic nucleus onwards, of the light information. In the present study we provide evidence that the endogenous release of GABA from suprachiasmatic nucleus terminals is implicated in the further transmission of light information to the pineal gland. Bilateral administration of the GABA-antagonist bicuculline to hypothalamic target areas of the suprachiasmatic nucleus completely prevents the inhibitory effect of nocturnal light on melatonin secretion and the present study thus documents that retina mediated photic activation of suprachiasmatic nucleus neurons induces the release of GABA from efferent suprachiasmatic nucleus nerve terminals, resulting in an inhibition of melatonin release by the pineal gland. Together with our previous (electro)physiological data these results identify GABA as an important mediator of rapid synaptic transmission of suprachiasmatic nucleus output to its target areas. PMID- 10366003 TI - Potentiation of excitotoxic injury by high concentrations of extracellular reduced glutathione. AB - Glutathione is present in the central nervous system in millimolar concentrations, and is a predominant intracellular antioxidant and detoxicant. In addition, glutathione is released into the extracellular space via a depolarization-enhanced process. Although the role of extracellular glutathione has not been precisely defined, a growing body of experimental evidence suggests that it has multifaceted electrophysiological effects. At low micromolar concentrations, glutathione depolarizes neurons by binding to its own receptors and modulates glutamatergic excitatory neurotransmission by displacing glutamate from its ionotropic receptors. At higher concentrations, reduced glutathione may increase N-methyl-D-aspartate receptor responses by interacting with its redox sites. In this study, the effect of extracellular glutathione on excitotoxic neuronal injury was quantitatively assessed in murine cortical cell cultures. Neuronal death due to 20-25 h exposure to 6-9 microM N-methyl-D-aspartate was not altered by 10-100 microM reduced glutathione but was markedly enhanced by 300 1000 microM reduced glutathione; kainate neurotoxicity was unaffected. Two related compounds that lack a sulfhydryl group, oxidized glutathione and S hexylglutathione, had no significant effect on N-methyl-D-aspartate neurotoxicity alone but completely blocked the effect of reduced glutathione. Mercaptoethanol, a sulfhydryl reducing agent that increases N-methyl-D-aspartate receptor responses by interacting with redox sites, increased N-methyl-D-aspartate neurotoxicity to a degree comparable to that of reduced glutathione; this effect was also blocked by equimolar S-hexylglutathione or oxidized glutathione. Addition of reduced glutathione to mercaptoethanol did not further increase N- methyl-D-aspartate-induced neuronal death. These results suggest that release of reduced glutathione from central nervous system cells that are subjected to traumatic or ischemic insults may enhance excitotoxic neuronal loss. Although multiple mechanisms may account for this phenomenon, the high concentrations required suggest that it is at least partly mediated by reduction of N-methyl-D aspartate receptor redox sites. PMID- 10366004 TI - Activity and expression of JNK1, p38 and ERK kinases, c-Jun N-terminal phosphorylation, and c-jun promoter binding in the adult rat brain following kainate-induced seizures. AB - The activity and/or expression of the mitogen-activated protein kinases c-Jun N terminal kinase 1, p38 and extracellular signal-regulated kinases 1/2, as well as their substrates, the transcription factors c-Jun and activating transcription factor-2, were examined following systemic application of kainate in the cortex and hippocampus of the adult rat brain. The protein expression levels of all three mitogen-activated protein kinases remained constant during the observation period. Unexpectedly, c-Jun N-terminal kinase 1 was the only mitogen-activated protein kinase activated in this model of excitotoxicity, its activity raised from between 1 and 3 h moderate basal to maximal levels between 6 and 12 h. In contradistinction, activity of extracellular signal-regulated kinases 1/2 fell from their substantial basal levels and did not recover; activity of p38 was characterized by a high basal level that almost entirely disappeared and did not return to basal levels even 10 days after kainate application. c-Jun protein was rapidly expressed, with a maximum after 3 h and a slow decline after 12 h. Supershift assays revealed that, during the early induction phase of the c-jun gene, the proximal activator protein-1 (jun1) site of the c-jun promoter was mainly occupied by the constitutively expressed activating transcription factor 2, whereas the late induction correlated with the predominant binding of c-Jun and, to a lesser extent, activating transcription factor-2 to the distal activator protein-1 (jun2) site. The time-course of the N-terminal phosphorylation of c-Jun as determined by immunocytochemistry paralleled the activity of c-Jun N-terminal kinase 1 and showed a compartment-specific regulation between 3 and 12 h. A second set of supershift experiments demonstrated that c-Jun, but not activating transcription factor 2, bound to activator protein-1 sites in the promoter of substance P and collagenase genes, but not of the cyclo-oxygenase-2 gene. Our results demonstrate that activation of c-Jun N-terminal kinase 1, phosphorylation of c-Jun and selective occupation of the c-jun promoter by activating transcription factor-2 or c-Jun are part of the neuronal response following excitotoxicity that is considered as the mechanism for neuronal apoptosis in vivo. Some of these findings differ substantially from in vitro experiments and underline the necessity to analyse the neuronal stress pathways in the adult brain. PMID- 10366005 TI - Growth-associated phosphoprotein expression is increased in the supragranular regions of the dentate gyrus following pilocarpine-induced seizures in rats. AB - Neuroplasticity has been investigated considering the neuronal growth-associated phosphoprotein as a marker of neuronal adaptive capabilities. In the present work, studying the hippocampal reorganization observed in the epilepsy model induced by pilocarpine, we carried out quantitative western blotting associated with immunohistochemistry to determine the distribution of growth-associated phosphoprotein in the hippocampus of rats in acute, silent and chronic periods of this epilepsy model. The fibers and punctate elements from the inner molecular layer of the dentate gyrus were strongly immunostained in animals killed 5 h after status epilepticus, compared with the same region in control animals. Rats presenting partial seizures showed no alterations in the immunostaining pattern compared with saline-treated animals. The hippocampal dentate gyrus of animals during the seizure-free period and presenting spontaneous recurrent seizures was also characterized by strong growth-associated phosphoprotein immunostaining of fibers and punctate elements in the inner molecular layer, contrasting with the control group. As determined by western blotting analysis, growth-associated phosphoprotein levels increased following status epilepticus and remained elevated at the later time-points, both during the silent period and during the period of chronic recurring seizures. Pilocarpine-treated animals, which did not develop status epilepticus, showed no change in growth-associated phosphoprotein levels, indicating that status epilepticus is important to induce growth associated phosphoprotein overexpression. The measurement of this overexpression could represent one of the early signals of hippocampal reorganization due to status epilepticus-induced damage. PMID- 10366006 TI - Reduction of lipopolysaccharide-induced neurotoxicity in mixed cortical neuron/glia cultures by femtomolar concentrations of pituitary adenylate cyclase activating polypeptide. AB - Stimulation of murine primary mixed cortical neuron/glia cultures with lipopolysaccharide, an endotoxin, was used as a model for inflammatory disorders of the central nervous system. Lipopolysaccharide (20 microg/ml) increased the secretion of lactate dehydrogenase, a marker for cell injury, and nitric oxide into the culture medium. The lipopolysaccharide-induced release of lactate dehydrogenase into the culture medium was reduced by pituitary adenylate cyclase activating polypeptide (PACAP) at 10(-14)-10(-12) M. The 27- and 38-amino-acid forms of PACAP were equipotent and their dose-response curves were U-shaped. PACAP6-38, a specific type I PACAP receptor antagonist, blocked the reduction by PACAP38 of the lipopolysaccharide-induced release of lactate dehydrogenase. The lipopolysaccharide-induced secretion of nitric oxide into the culture medium was reduced by PACAP at 10(-14)-10(-12) M and 10(-8)-10(-6) M. The 27- and 38-amino acid forms of PACAP were equipotent. PACAP6-38 blocked the reduction of the lipopolysaccharide-induced secretion of nitric oxide by PACAP38 at 10(-12) M, but not at 10(-8) M. Vasoactive intestinal polypeptide reduced the lipopolysaccharide induced release of lactate dehydrogenase into the culture medium at 10(-14)-10( 12) M, but these concentrations of vasoactive intestinal polypeptide had no effect on the lipopolysaccharide-induced secretion of nitric oxide. PACAP6-38 did not effect the reduction of the lipopolysaccharide-induced release of lactate dehydrogenase into the culture medium by 10(-12) M vasoactive intestinal polypeptide. These results indicate that stimulation of type I PACAP receptors by femtomolar concentrations of PACAP can prevent neuron death in a model for inflammatory disorders of the CNS. These results suggest that PACAP is also an extraordinarily potent inhibitor of some microglial functions. PMID- 10366007 TI - Neuroprotective effect of mild hypothermia cannot be explained in terms of a reduction of glutamate release during ischemia. AB - An exogenous glutamate injection into the hypothermic hippocampal CA1 during 5 min ischemia produced the same extent of extracellular glutamate levels as observed in the normothermic CA1 during 5-min ischemia; however, neuronal death was not induced in the hypothermic CA1. Glutamate is released excessively into the extracellular space during ischemia, and is thought to induce brain injury by its neurotoxicity. It has been reported that the massive glutamate release is reduced by mild hypothermia, and it has been proposed that the reduction of ischemia-induced glutamate release exerts the neuroprotective effect on postischemic neuronal death. In the present study, to determine whether the neuroprotective effect of mild hypothermia on postischemic hippocampal CA1 neuronal death is due to the reduction of ischemia-induced glutamate release, gerbils were subjected to 5-min ischemia under hypothermic condition at 31 degrees C and were simultaneously injected exogenously with L-glutamate, so that the hypothermic CA1 around a microdialysis probe was exposed to the same extracellular glutamate levels as seen during normothermic ischemia, and the histological outcome was examined. An injection with 1 mM L-glutamate into the hypothermic CA1 during 5-min ischemia produced a similar extent of increased glutamate (17-fold increase) to that observed in the normothermic CA1 during 5 min ischemia (16-fold increase). However, neuronal death was not induced in the hypothermic CA1. This result indicates that the neuroprotective effect of mild hypothermia cannot be explained in terms of a reduction of glutamate release during ischemia. PMID- 10366008 TI - Effect of hypoxia on membrane potential and resting conductance in rat hippocampal neurons. AB - The present patch-clamp study describes the effect of hypoxia at 30-31 degrees C on membrane potential and resting conductance in pyramidal cells from the hippocampal CA1 region in rat brain slices. The initial effect of hypoxia was a gradual hyperpolarization; the peak change in membrane potential measured over 15 min was -5.3 +/- 0.22 mV (P < 0.0001). After reoxygenation followed a transient hyperpolarization measuring -1.8 +/- 0.24 mV (P < 0.0001) and a subsequent normalization of the membrane potential, which after 5 min did not differ from its level prior to the hypoxic episode. Voltage-clamp analysis showed that the hypoxic hyperpolarization was related to an outward current at the holding potential (-60 mV) and an increase in resting conductance. The effect was not influenced by intracellular Cl- concentration, which indicated that it was not due to an inward flow of Cl- ions. The addition of tolbutamide, glibenclamide and dantrolene sodium did not affect the hypoxic hyperpolarization, neither did the presence of ATP in the pipette solution. The presence/absence of glucose in the perfusion medium did not influence the initial hyperpolarization during hypoxia; however, glucose seemed to prevent the subsequent depolarization under hypoxia. It was concluded that hypoxia caused an initial hyperpolarization of CA1 cells which was related to an increase in the resting conductance. The results did not suggest the involvement of ATP-sensitive K+ channels. PMID- 10366009 TI - Neural transplantation of human neuroteratocarcinoma (hNT) neurons into ischemic rats. A quantitative dose-response analysis of cell survival and behavioral recovery. AB - Transplantation of fetal neuronal tissue has been used successfully to ameliorate symptoms of neurodegenerative disease in animals and humans. This technique has recently been extended as an experimental treatment for ischemic brain damage. However, due to ethical issues with the use of fetal cells for the treatment of any human disease, there has been a concerted effort to find alternative graft sources for neural transplantation. The human neuroteratocarcinoma neuron cell is derived from an embryonal teratocarcinoma cell line that can be differentiated into post-mitotic neurons. Neural transplantation of human neuroteratocarcinoma neurons has recently been shown to produce behavioral amelioration of symptoms in rats with ischemia-induced injury. The present study was undertaken to: (i) determine the minimum effective number of transplanted human neuroteratocarcinoma neurons required for amelioration of ischemia-induced behavioral dysfunction; and (ii) quantify the survival of human neuroteratocarcinoma neurons in vivo. Transplants of 0, 5, 10, 20, 40, 80 or 160 x 10(3) human neuroteratocarcinoma neurons were made into rats that sustained ischemic damage. Animals that received 40, 80 or 160 x 10(3) human neuroteratocarcinoma neurons demonstrated a dose dependent improvement in performance of both the passive avoidance and elevated body swing tests. At the conclusion of behavioral testing, human neuroteratocarcinoma neurons were identified in paraffin sections with human neural cell adhesion molecule MOC-1 and human neurofilament antibodies. Transplants of 80 or 160 x 10(3) human neuroteratocarcinoma neurons demonstrated a 12-15% survival of human neuroteratocarcinoma neurons in the graft, while transplants of 40 x 10(3) human neuroteratocarcinoma neurons demonstrated a 5% survival. Transplantation of human neuroteratocarcinoma neurons ameliorated behavioral deficits produced by ischemic damage. The human neuroteratocarcinoma neuron, additionally, showed greater survival than that reported for fetal cells when transplanted into the brain. Therefore, this readily available cell may prove to be an excellent candidate for the treatment of ischemic damage in human patients. PMID- 10366010 TI - Opposite modulation of cortical N-methyl-D-aspartate receptor-mediated responses by low and high concentrations of dopamine. AB - To examine whether dopamine modulates cortical N-methyl-D-aspartate receptor mediated glutamate transmission, whole-cell recordings were made from identified pyramidal cells located in layers V and VI of the medial prefrontal cortex of the rat using a slice preparation. In the presence of tetrodotoxin and the absence of Mg2+, a brief local application of N-methyl-D-aspartate evoked an inward current which was blocked by the N-methyl-D-aspartate antagonist dizocilpine maleate but not affected by the non-N-methyl-D-aspartate antagonist 2,3-dihydroxy-6-nitro-7 sulfamoyl-benzo(f)quinoxaline, suggesting that the observed current is mediated by N-methyl-D-aspartate receptors located on recorded cells. Bath application of dopamine produced opposite effects on the N-methyl-D-aspartate current depending on the concentrations of dopamine applied. At low concentrations (<50 microM), dopamine enhanced the N-methyl-D-aspartate current, whereas at higher concentrations, dopamine suppressed the current. The same concentrations of dopamine did not significantly affect the inward current induced by the non-N methyl-D-aspartate agonist alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid. The enhancing effect of dopamine on the N-methyl-D-aspartate response was mimicked by the D1 agonist SKF38393 and blocked by the D1 antagonist SCH31966, whereas the suppressing effect was mimicked by the D2 agonist quinpirole and blocked by the D2 antagonist eticlopride. The above results suggest that dopamine at low concentrations acts preferentially on D1-like receptors to promote N methyl-D-aspartate receptor-mediated transmission, while at high concentrations dopamine also activates D2-like receptors, leading to a suppression of the N methyl-D-aspartate function. This differential modulation of N-methyl-D-aspartate function may have significant implications for understanding behaviors and disorders involving both cortical dopamine- and glutamate-mediated neurotransmission. PMID- 10366011 TI - Regulation of human D1 dopamine receptor function and gene expression in SK-N-MC neuroblastoma cells. AB - SK-N-MC human neuroblastoma cells express functional D1, but not D5, dopaminergic receptors. Stimulating cells with dopamine or the D1-selective agonist, SKF R 38393, rapidly (t(1/2) = 1 h) resulted in > 95% attenuation of dopamine-mediated accumulation of cyclic AMP, without any change in D1 dopamine receptor levels. Prolonged (> 4 h) exposure of cells to dopamine attenuated D1 receptor levels to 45-50% of control (t(1/2) = 8 h) and was accompanied by a loss of high-affinity binding sites. At the molecular level, the expression of D1 receptor messenger RNA was bimodal: an initial increase (by approximately 60%) of receptor messenger RNA within 2 h of treatment of cells with dopamine was followed by a decline to 50% below control messenger RNA levels. Low concentrations (1-10 nM) of dopamine also potentiated D1 messenger RNA levels (up to 48%), resulting in a twofold increase in receptor levels. Transfection studies with the cloned human D1 promoter construct, pGL-D1P, indicated that the up-regulation of D1 messenger RNA was due to activation of promoter by dopamine. The dopamine-mediated up regulation of both D1 receptor messenger RNA and promoter was prevented by the D1 selective antagonist, SCH 23390. The results suggest that dopamine regulates D1 receptor gene and protein expression in a bimodal manner, partly through activation of the receptor promoter. Moreover, the effects of dopamine are independent of the second messenger, cyclic AMP. PMID- 10366012 TI - Expression of dopamine receptors in the subthalamic nucleus of the rat: characterization using reverse transcriptase-polymerase chain reaction and autoradiography. AB - We analysed the expression of dopamine receptor subtypes in the subthalamic nucleus by means of reverse transcriptase-polymerase chain reaction. We also studied, using autoradiography, all pharmacologically characterized dopamine receptors in four subregions of the subthalamic nucleus. For comparison, dopamine receptor subtypes were also evaluated in brain regions where they are more abundant and well characterized. The radioligands used were: [3H]SCH-23390, [3H]emonapride and [3H]2-dipropylamino-7-hydroxy-1,2,3,4-tetrahydronaphthalene for dopamine D1, D2 and D3 receptors, respectively; and [3H]YM-09151-2 in the presence of raclopride for dopamine D4 receptors. Finally, we also evaluated the effect of unilateral 6-hydroxydopamine injection into the medial forebrain bundle on dopamine receptor levels expressed in the ipsilateral subthalamic nucleus. The lesion was estimated by decrease in the binding of [3H]WIN-35428, a specific dopamine transporter label. D1, D2 and D3 receptor messenger RNAs and binding sites were present in the subthalamic nucleus, but no messenger RNA for D4 receptors was found, although specific binding sites for these receptors were observed. As compared to the intact side, the 6-hydroxydopamine lesion did not change D1 receptors, increased D2 receptors, and decreased D3 receptors and the dopamine transporter. The results suggest that postsynaptic D1, D2 or D3 receptors can mediate the effect of dopamine on subthalamic nucleus neuronal activity. D4 receptors would mediate exclusively presynaptic effects. These results reinforce the idea that dopamine receptors in the subthalamic nucleus may play an important role in the physiology of the basal ganglia and in the pathophysiology of Parkinson's disease. PMID- 10366014 TI - Serotonin hyperinnervation in the adult rat ventral mesencephalon following unilateral transection of the medial forebrain bundle. Correlation with reactive microglial and astroglial populations. AB - We have previously studied changes in the serotoninergic and dopaminergic nigrostriatal systems following transection of the medial forebrain bundle and found a long-term axotomy-induced increase in the levels of serotonin and its main metabolite, 5-hydroxyindolacetic acid in substantia nigra [Venero et al. (1997) J. Neurochem. 68, 2458-2468]. In an attempt to find a rationale for this effect, we have performed an immunohistochemical study. Transection of the medial forebrain bundle of the rat interrupted most of the ascending serotoninergic pathways from the raphe nuclei as revealed by serotonin immunoreactivity. While serotonin immunostaining was almost absent in striatum, it doubled in the ventral mesencephalon at 21 days postlesion. This axotomy-induced increase was accompanied by an increased density of the serotonin nerve terminal network in the ipsilateral substantia nigra and ventral tegmental area. The increase in serotonin immunoreactivity was in line with the measured levels of serotonin and 5-hydroxyindolacetic acid in substantia nigra. In addition, the distribution pattern of glial fibrillary acidic protein-immunoreactive astrocytes and OX42 immunoreactive microglia correlated highly with the location of increased serotonin fibre density in the ventral mesencephalon, especially in ventral tegmental area and in the most medial part of substantia nigra. We suggest that a pruning effect may underly the axotomy-induced increase in serotonin immunoreactivity in the ventral mesencephalon, and further, that activated astroglia and microglia may play a role in directing serotoninergic axonal regeneration following axotomy. PMID- 10366013 TI - Presynaptic muscarinic (M3) receptors reduce excitatory transmission in dopamine neurons of the rat mesencephalon. AB - The effects of carbachol (0.01-30 microM) and muscarine (10-30 microM) on the excitatory synaptic potentials were studied using conventional intracellular recordings from dopaminergic neurons in rat mesencephalic slices. Both muscarinic agonists reversibly reduced the excitatory synaptic potentials, evoked by local electrical stimulation. The EC50 for carbachol was determined to be 4.5 microM. The maximal degree of the excitatory synaptic potentials suppression caused by carbachol and muscarine was around 40% of control. This suppression was completely blocked by the non-specific muscarinic antagonist atropine (1 microM) and the selective M3 antagonist 4-diphenylacetoxy-N-methylpiperidine methiodide (1 microM). Other antagonists, preferentially acting at M1, M2 and M4 receptors, were not effective. Furthermore, the acetylcholinesterase inhibitor, physostigmine (50 microM), decreased the amplitude of the excitatory synaptic potentials, indicating that ambient acetylcholine can depress this potential. Direct depolarizing responses to glutamate were not changed by muscarine. In addition, muscarine facilitated the second excitatory synaptic potentials during a paired-pulse protocol. Thus, the effect of the muscarinic agonists is attributable to a presynaptic locus of action. The action of muscarine was not mediated by an N-ethylmaleimide-sensitive G-protein since it was not modified by a treatment of the slices with this agent. The calcium channels blockers, omega conotoxin GIVA, omega-agatoxin IVA and omega-conotoxin MVIIC did not affect the action of muscarine on the excitatory synaptic potentials. When the potassium currents were reduced by extracellular barium and 4-aminopyridine, the muscarinic agonists still depressed the excitatory synaptic potentials. Our data indicate that presynaptically located M3 receptors modulate the excitatory transmission to midbrain dopaminergic neurons via a N-ethylmaleimide-insensitive G-protein which activates mechanisms neither linked to N-, P-, Q-type calcium channels nor to barium- and 4-aminopyridine-sensitive potassium channels. PMID- 10366015 TI - Time-course and localization of transferrin receptor expression in the substantia nigra of 6-hydroxydopamine-induced parkinsonian rats. AB - Parkinson's disease is a neurodegenerative disease characterized by dopaminergic cell death in the substantia nigra. The cause of the cell death is, however, obscure. Recently, accumulation of iron in the parkinsonian substantia nigra and iron-catalysed free radical generation have been proposed as possible causes of nigral cell death. The transferrin receptor has been implicated as a possible mediator of this iron accumulation in the parkinsonian substantia nigra. The present study investigated the distribution of transferrin receptor immunoreactive proteins and its co-localization with tyrosine hydroxylase in the normal rat substantia nigra and their expressions in the parkinsonian substantia nigra from three days to three months after 6-hydroxydopamine lesioning. Computer image analysis of the grey mean of transferrin receptor staining in the microvessels was also employed. The results showed that the transferrin receptor immunolabelling was localized in some neurons and glial cells in the normal substantia nigra pars compacta and pars reticulata, and that about 54% of tyrosine hydroxylase-positive cells were also stained with transferrin receptor. There was a decrease of tyrosine hydroxylase- and transferrin receptor-positive cells in the 6-hydroxydopamine-lesioned substantia nigra. The grey mean of transferrin receptor staining in microvessels in the lesioned substantia nigra was, however, not different from that in the control. It was concluded that transferrin receptors in neurons, glial cells and microvessels might not be responsible for iron accumulation in the parkinsonian substantia nigra. The loss of transferrin receptor-immunopositive cells might, however, partly be accounted for by the death of transferrin receptor-positive dopaminergic cells induced by 6 hydroxydopamine lesioning. PMID- 10366016 TI - Selective blockade of serotonin-2C/2B receptors enhances mesolimbic and mesostriatal dopaminergic function: a combined in vivo electrophysiological and microdialysis study. AB - Electrophysiological techniques and in vivo microdialysis were used to investigate the relative contribution of central serotonin-2C/2B and serotonin-2A receptor subtypes in the control of mesolimbic and nigrostriatal dopaminergic function. Thus, extracellular single-unit recordings were performed from neurochemically identified dopamine neurons in the ventral tegmental area and the substantia nigra pars compacta, as well as simultaneous monitoring of accumbal and striatal basal dopamine release in anesthetized rats following the administration of serotonin-2C/2B (SB 206553), serotonin-2A (SR 46349B) or serotonin-2A/2B/2C (ritanserin) antagonists. Administration of SB 206553 (40-160 microg/kg, i.v.) caused a dose-dependent increase in the basal firing rate of ventral tegmental area and nigral dopamine neurons, reaching its maximum (45.2 and 28.5%, respectively) following 160 microg/kg. Moreover, burst activity was significantly enhanced by SB 206553 in the ventral tegmental area only. In contrast, injection of SR 46349B (40-160 microg/kg, i.v.), and ritanserin (40-160 microg/kg, i.v.) did not cause any significant change in the basal activity of these neurons. Basal dopamine release was significantly enhanced in both the nucleus accumbens (42%) and the striatum (33%) following the intraperitoneal administration of 5 mg/kg SB 206553. In contrast, SR 46349B (0.5 mg/kg, s.c.) and ritanserin (0.63 mg/kg, i.p.) failed to affect basal dopamine output in both regions. Taken together, these data indicate that the central serotonergic system exerts a tonic inhibitory control of mesolimbic and nigrostriatal dopaminergic pathway activity and that the serotonin-2C/2B receptor subtypes are involved in this effect. Moreover, these findings might open new possibilities for the employment of serotonin-2C/2B receptor antagonists in the treatment of neuropsychiatric disorders related to a hypofunction of central dopaminergic neurons. PMID- 10366017 TI - Clozapine and other 5-hydroxytryptamine-2A receptor antagonists alter the subcellular distribution of 5-hydroxytryptamine-2A receptors in vitro and in vivo. AB - In this study, we demonstrate that clozapine and other atypical antipsychotic drugs induce a paradoxical internalization of 5-hydroxytryptamine-2A receptors in vitro and a redistribution of 5-hydroxytryptamine-2A receptors in vivo. We discovered that clozapine, olanzapine, risperidone and the putative atypical antipsychotic drug MDL 100,907 all induced 5-hydroxytryptamine-2A receptor internalization in fibroblasts stably expressing the 5-hydroxytryptamine-2A receptor in vitro. Two 5-hydroxytryptamine-2A antagonists (mianserin and ritanserin), which have been demonstrated to reduce negative symptoms in schizophrenia, also caused 5-hydroxytryptamine-2A receptor internalization. Four different drugs, each devoid of 5-hydroxytryptamine-2A antagonist activity, had no effect on the subcellular distribution of 5-hydroxytryptamine-2A receptors in vitro. Treatment of rats for seven days with clozapine induced an increase in intracellular 5-hydroxytryptamine-2A receptor-like immunoreactivity in pyramidal neurons, while causing a decrease in labeling of apical dendrites in the medial prefrontal cortex. This redistribution of 5-hydroxytryptamine-2A receptors in pyramidal neurons was also seen when rats were chronically treated with another atypical antipsychotic drug, olanzapine. The typical antipsychotic drug haloperidol, however, did not induce a redistribution of 5-hydroxytryptamine-2A receptors in pyramidal neurons in the medial prefrontal cortex. Taken together, these results demonstrate that several atypical antipsychotic drugs with high 5 hydroxytryptamine-2A receptor affinities induce a redistribution of 5 hydroxytryptamine-2A receptors both in vivo and in vitro. It is conceivable that the loss of 5-hydroxytryptamine-2A receptors from the apical dendrites of pyramidal neurons is important for the beneficial effects of atypical antipsychotic drugs and other 5-hydroxytryptamine-2A antagonists in schizophrenia. PMID- 10366018 TI - Blockade of cannabinoid receptors by SR141716 selectively increases Fos expression in rat mesocorticolimbic areas via reduced dopamine D2 function. AB - The present study investigated, in rats, whether blockade of cannabinoid CB1 receptors may alter Fos protein expression in a manner comparable to that observed with antipsychotic drugs. Intraperitoneal administration of the selective CB1 receptor antagonist, SR141716, dose-dependently (1.0, 3.0 and 10 mg/kg) increased Fos-like immunoreactivity in mesocorticolimbic areas (prefrontal cortex, ventrolateral septum, shell of the nucleus accumbens and dorsomedial caudate-putamen), while motor-related structures such as the core of the nucleus accumbens and the dorsolateral caudate-putamen were unaffected. In the ventrolateral septum, taken as a representative structure, the Fos-inducing effect of SR141716 (10 mg/kg) was maximal 2 h after injection and returned to near control levels by 4 h. Within the prefrontal cortex, SR141716 increased the number of Fos-positive cells predominantly in the infralimbic and prelimbic cortices, presumptive pyramidal cells being the major cell types in which Fos was induced. The D1-like receptor antagonist, SCH23390 (0.1 mg/kg), did not prevent the Fos-inducing effect of SR141716 in any brain region examined (prefrontal cortex, nucleus accumbens, ventrolateral septum and dorsomedial caudate-putamen), although SCH23390 significantly reduced Fos expression induced by cocaine (20 mg/kg) in all these regions. By contrast, the dopamine D2-like agonist, quinpirole (0.25 mg/ kg), counteracted SR141716-induced Fos-like immunoreactivity in the ventrolateral septum, the nucleus accumbens and the dorsomedial caudate putamen, while no antagonism was observed in the prefrontal cortex. Microdialysis experiments in awake rats indicated that SR141716, at doses which increased Fos expression (3 and 10 mg/kg), did not alter dopamine release in the shell of the nucleus accumbens. Finally, SR141716 increased the levels of neurotensin-like immunoreactivity in the nucleus accumbens, but not in the caudate-putamen. Collectively, the present results show that blockade of cannabinoid receptors increases Fos- and neurotensin-like immunoreactivity with characteristics comparable to those reported for atypical neuroleptic drugs. PMID- 10366019 TI - Nitrergic stimulation of the locus coeruleus modulates blood pressure and heart rate in the anaesthetized rat. AB - To investigate whether nitric oxide is involved in the cardiovascular responses mediated via the locus coeruleus, the effects of microinjections of L-arginine and L-glutamate into the locus coeruleus on blood pressure and heart rate were investigated in sodium pentobarbitone-anaesthetized rats. Unilateral microinjection of L-arginine (25, 50 nmol) elicited dose-related depressor (-17 +/- 4, -25 +/- 4 mmHg) and bradycardic (13 +/- 3, 24 +/- 6 b.p.m.) effects. Furthermore, these effects were attenuated by prior local microinjection of N(G) nitro-L-arginine (40 nmol). Peripheral muscarinic receptor blockade with atropine methyl nitrate (1 mg/kg, i.v.) attenuated the bradycardic but not the depressor responses to L-arginine. L-Glutamate (2 nmol) microinjections also mediated depressor (-27 +/- 6 mmHg) and bradycardic (53 +/- 23 b.p.m.) effects that were attenuated by microinjections of dizocilpine maleate (1 nmol) into the locus coeruleus. In addition, pretreatment with N(G)-nitro-L-arginine (40 nmol) also significantly attenuated the depressor response elicited by L-glutamate. These results suggest that nitrergic and glutamatergic pathways are operative within the locus coeruleus to modulate cardiovascular function, and also that a functional interaction may exist between the nitrergic and glutamatergic systems within the rat locus coeruleus. PMID- 10366020 TI - Effects of ammonia in vitro on endogenous taurine efflux and cell volume in rat cerebrocortical minislices: influence of inhibitors of volume-sensitive amino acid transport. AB - Rat cerebrocortical minislices were incubated with physiological saline in the absence or presence of 5 mM ammonium acetate ("ammonia") and/or inhibitors of osmosensitive amino acid transport: 50 microM niflumic acid and 100 microM N ethyl-maleimide for 60 min, with medium changes after 20 min and 40 min. The efflux of endogenous taurine, glutamate and glutamine was assayed by high performance liquid chromatography, and steady-state cell volumes were monitored in the slices with the [14C]inulin method. In the absence of ammonia, niflumic acid abolished taurine efflux but did not affect glutamate or glutamine efflux at all time-points, and increased cell volume at 20 min and 60 min. N-Ethyl maleimide increased taurine, glutamine and glutamate efflux at 20 min and 40 min, inhibited taurine and glutamine efflux at 60 min, and increased cell volume at 20 min. Ammonia strongly stimulated taurine (by 380% at 20 min), and only moderately glutamate (30% at 20 min) or glutamine efflux (76% at 20 min). Ammonia increased cell volume above the control level at all time-points. Niflumic acid inhibited, but did not abolish ammonia-dependent taurine and glutamine efflux, and did not change glutamate efflux. The effects of ammonia + niflumic acid on cell volume did not differ from the effects of each compound separately. N-Ethyl-maleimide inhibited ammonia-dependent efflux of all three amino acids except for stimulation of glutamate efflux at 20 min. N-Ethyl-maleimide + ammonia decreased the cell volumes more than did each compound separately. It is concluded that although ammonia-induced taurine efflux is accompanied by an increase in cell volume, the underlying mechanism is not simply a cell volume regulatory response normally observed in hypoosmotic stress. Increased efflux of taurine, which is an inhibitory amino acid and a cell membrane protectant, may serve to counteract the deleterious effects of increased excitatory transmission accompanying acute hyperammonemic insult. PMID- 10366021 TI - Expression of alpha1D adrenergic receptor messenger RNA in oxytocin- and corticotropin-releasing hormone-synthesizing neurons in the rat paraventricular nucleus. AB - The paraventricular nucleus of the hypothalamus contains a number of intermingled populations of neuroendocrine cell groups involved in the hormonal stress response, including cells synthesizing corticotropin-releasing hormone and oxytocin. Ascending noradrenergic afferents to the paraventricular nucleus, acting through alpha1 adrenergic receptors, are thought to play a role in stress induced activation of the hypothalamic-pituitary-adrenal axis. We have previously demonstrated that, of the three known alpha1 adrenergic receptor subtypes, messenger RNA for the alpha1D subtype is the most prominently expressed in the paraventricular nucleus. Thus, regulation of the expression of this receptor may be important in modulation of the stress response. It is currently unknown, however, which populations of stress-related neuroendocrine cells in the paraventricular nucleus express alpha1 receptors, or whether the excitatory influence of norepinephrine in stress is exerted directly on neurons expressing oxytocin or corticotropin-releasing hormone. Thus, in the present study, we used dual in situ hybridization, combining a digoxigenin-labeled riboprobe encoding the rat alpha1D adrenergic receptor with radiolabeled riboprobes for oxytocin or corticotropin-releasing hormone, to determine the degree to which these neurons in the paraventricular nucleus express alpha1D adrenergic receptors. In sections through the rostral and mid-level paraventricular nucleus, nearly all (>95%) oxytocin neurons also expressed alpha1D messenger RNA. In contrast, the populations of corticotropin-releasing hormone- and alpha1D-expressing cells overlapped only partially, with most alpha1D expression situated more laterally. A subset (37%) of the neurons expressing corticotropin-releasing hormone also expressed alpha1D messenger RNA, and these were found almost entirely within the region of overlap in the lateral aspect of the medial parvocellular region. These observations support a direct role for alpha1 receptors in regulation of oxytocin secretion. Expression of alpha1D messenger RNA in distinct subsets of cells synthesizing corticotropin-releasing hormone may also help to clarify contradictory and inconsistent observations in the literature regarding the role of norepinephrine in the stress response, and may account for a presumed stressor specific role for norepinephrine in activation of the hypothalamic-pituitary adrenal axis. PMID- 10366022 TI - Developmental expression of alpha-synuclein in rat hippocampus and cerebral cortex. AB - Alpha-synuclein is an evolutionary highly conserved neuronal protein localized in presynaptic nerve terminals. The protein has been suggested to be involved in the pathogenesis of neurodegenerative diseases, but little is known about the physiological function of the protein. In the present study we used newborn, three, 14, 93 and 710-day-old rats to examine the expression of alpha-synuclein messenger RNA and protein during development of the hippocampus and cerebral cortex. Using in situ hybridization and an S1 nuclease protection assay, we found a high expression of alpha-synuclein messenger RNA during early postnatal development, followed by a marked decrease between postnatal days 14 and 93. In contrast, the amount of alpha-synuclein protein, as determined by immunoblotting, continued to increase throughout development and remained at a high level for at least two years. The persistent high expression of alpha-synuclein protein throughout development suggests that the protein is involved in maintaining synaptic function. Furthermore, the discrepancy between the levels of alpha synuclein messenger RNA and protein after postnatal day 14 indicates that the amount of alpha-synuclein is determined by post-transcriptional regulation, and not by messenger RNA expression alone. To estimate the changes of alpha-synuclein expression per synapse, we compared the developmental expression of alpha synuclein with synaptophysin, a well-established synaptic marker. The alpha synuclein/synaptophysin messenger RNA and protein ratio was high during early development, but low in adult (postnatal day 93) and old (postnatal day 710) rats. This could indicate a higher expression of alpha-synuclein per synapse during early development. PMID- 10366023 TI - Neuronal localization of the Adenomatous polyposis coli tumor suppressor protein. AB - Recent biochemical studies have demonstrated that the adenomatous polyposis coli gene, initially identified via its link to colon cancer, is expressed at high levels in the brain. Furthermore, the ability of this tumor suppressor protein to bind to Discs-Large and beta-catenin, proteins implicated in organizing synaptic structure, point to a role for APC in neuronal signalling. However, anatomical studies have provided conflicting results regarding its localization in brain. In situ hybridization studies predict neuronal expression of APC, while immunostaining studies performed with a panel of N-terminal antibodies detected staining of glial cells, especially oligodendrocytes. In this study, we have examined the basis for this discrepancy and provide evidence that the glial staining pattern detected in previous studies reflects cross-reactivity with an unrelated antigen rather than the localization of APC. Furthermore, we have performed immunohistochemical studies with a C-terminal APC antibody which reveal a neuronal pattern of staining closely matching that predicted by the in situ studies. For example, in the hippocampus APC immunostaining is detected in the pyramidal neurons and dentate granule cells, which fits well with the localization of APC mRNA. Examination of APC immunostaining in other regions revealed that particularly intense staining was displayed by large neurons, including layer V cortical pyramidal neurons, cerebellar Purkinje cells, and olfactory bulb mitral cells. Within labeled neurons, APC staining was apparent in the cytoplasm, as well as in dendritic and axonal processes. To help clarify the localization of APC in brain, we have conducted additional in situ hybridization and immunohistochemical studies. These results provide compelling evidence that APC is expressed predominantly in neurons rather than in glial cells as reported previously. PMID- 10366024 TI - Differential expression of bcl-w and bcl-x messenger RNA in the developing and adult rat nervous system. AB - The bcl-2 family of proteins comprises both anti-apoptotic and pro-apoptotic members, which play a pivotal role in regulating cell death. Bcl-w is a recently identified member of this family, which was shown to inhibit apoptosis in haemopoietic cell lines. However, the function and expression patterns of bcl-w in the nervous system have so far not been described. We have cloned complementary DNA corresponding to rat bcl-w and analysed the expression of bcl-w messenger RNA during rat brain development, using RNA blotting and in situ hybridization techniques. We also compared the expression patterns of bcl-w with those of bcl-xL. During brain development, the levels of bcl-w messenger RNA were found to increase, with highest expression located to specific regions of the mature brain, such as hippocampus, cerebellum, piriform cortex and locus coeruleus. Bcl-w messenger RNA was expressed by neurons, as shown with double labeling with neuronal markers. In contrast to bcl-w, bcl-xL messenger RNA expression levels were highest during early development, particularly in cortex, hippocampus, thalamus, spinal cord and dorsal root ganglia. During postnatal development the expression of bcl-xL messenger RNA decreased and were only detected at low levels in the adult nervous system. As shown earlier for bcl-2, the expression of bcl-w and bcl-x messenger RNA in cultured cerebellar granule cells was not altered by the deprivation of neurotrophic factors. The present results suggest that bcl-w may play an important role in the mature nervous system. PMID- 10366025 TI - Localization of neuroendocrine secretory protein 55 messenger RNA in the rat brain. AB - Neuroendocrine secretory protein 55 (NESP55) is a recently characterized secretory protein localized to large dense-core vesicles resembling the class of chromogranins. We investigated the distribution of the messenger RNA encoding for NESP55 in the rat brain by in situ hybridization with specific 35S-labelled oligonucleotides. NESP55 messenger RNA was detected only on neuronal but not glial cells. In the brain, expression of NESP55 messenger RNA was most prominent in several areas throughout the midbrain and brainstem, including the locus coeruleus, the raphe complex and the reticular formation. NESP55 messenger RNA expressing cells were also found in many areas and nuclei throughout the hypothalamus. Neocortical areas, the hippocampus and the cerebellum were devoid of NESP55 messenger RNA-containing neurons. From this distribution pattern, a significant overlap of NESP55 expression with the noradrenergic, adrenergic and serotonergic transmitter systems was evident. The present study defines, for the first time, the cellular localizaton of NESP55 messenger RNA in the rat brain. The present results provide the basis for future studies defining the as yet obscure function of NESP55. PMID- 10366026 TI - Distribution of synaptosomal-associated protein 25 in nerve growth cones and reduction of neurite outgrowth by botulinum neurotoxin A without altering growth cone morphology in dorsal root ganglion neurons and PC-12 cells. AB - Synaptosomal-associated protein 25 has been regarded as one of the target associated soluble N-ethylmaleimide-sensitive fusion attachment protein receptors essential for exocytosis of vesicles in synapses. We have previously reported that cleavage of syntaxin, which is another target-associated soluble N ethylmaleimide-sensitive fusion attachment protein receptor, with botulinum neurotoxin C1 resulted in inhibition of neurite extension and morphological changes including growth cone collapse and large vacuole formation. As an attempt to explore the mechanism of growth cone extension, we examined the ultrastructural localization of synaptosomal-associated protein 25 in growth cones with or without treatment of botulinum neurotoxin A, which cleaves synaptosomal-associated protein 25. In dorsal root ganglion neurons, light microscopy demonstrated synaptosomal-associated protein 25 immunoreactivity throughout the neurons, including the cell bodies, neurites and growth cones. Using electron microscopy, gold signals immunoreactive for synaptosomal associated protein 25 were identified diffusely in the cytoplasm of the growth cones. In contrast, in PC-12 cells, a large number of gold signals were localized on the plasma membranes. High levels of signal were also found in the cytoplasm in the central region of the growth cones. We also confirmed that botulinum neurotoxin A treatment reduced neurite extension by about 50%. However, both in dorsal root ganglion neurons and in PC-12 cells we found no differences in the ultrastructure nor in the localization of synaptosomal-associated protein 25 between growth cones with and without toxin treatment. These results indicate that cleavage of synaptosomal-associated protein 25 inhibits growth cone extension in a manner different than that of syntaxin cleavage. The results of this study suggest the possibility that synaptosomal-associated protein 25 is involved in growth cone extension through a process independent of vesicle fusion. PMID- 10366027 TI - Effects of axotomy on the expression and ultrastructural localization of N cadherin and neural cell adhesion molecule in the quail ciliary ganglion: an in vivo model of neuroplasticity. AB - Postganglionic nerve crush of the avian ciliary ganglion induces detachment of preganglionic terminals from the soma of the injured ciliary neurons, followed by reattachment at about the same time that the postganglionic axons regenerate to their targets. In order to determine the role played by cell adhesion molecules in this response, we have studied injury-induced changes in the amount and distribution of N-cadherin and neural cell adhesion molecule, together with modifications in the expression of their messenger RNAs. Both N-cadherin and neural cell adhesion molecule immunoreactivities associated with postsynaptic specializations decreased between one and three days following postganglionic nerve crush, preceding the detachment of the preganglionic boutons. Immunoreactivities subsequently increased between 13 and 20 days, in parallel with restoration of synaptic contacts on the ganglion cells and the progressive reinnervation of the peripheral targets. In contrast to the rapid decrease in immunoreactivity, the messenger RNA levels of N-cadherin and neural cell adhesion molecule both increased after crush, and remained elevated throughout the 20-day period of the experiment. These results are consistent with roles for N-cadherin and neural cell adhesion molecule in the maintenance of synaptic contacts. The rapid regulation of these proteins in injury-induced synaptic plasticity occurs at the post-transcriptional level, whereas longer term regulation associated with the re-establishment of synapses may be promoted by the increased levels of gene expression. PMID- 10366028 TI - A role for spinal lamina I neurokinin-1-positive neurons in cold thermoreception in the rat. AB - Lamina I neurons of the spinal cord convey specific nociceptive activity to the brain. A subpopulation of lamina I cells bears substance P receptors (neurokinin 1) and recent studies have shown that these neurons encode for the intensity of noxious peripheral stimulation. Here, we report that cool thermal stimuli, applied to the hindpaw of anaesthetized rats, induce Fos expression in lamina I neurokinin-1 neurons that is graded with respect to the intensity of the thermal stimulus. Thus, as the temperature of the stimulus was reduced, both the total number of neurokinin-l-positive neurons expressing Fos and the proportion of Fos nuclei present within neurokinin-1 cells showed a significant increase. These data show that lamina I neurokinin-1 cells encode the intensity of noxious cooling of the skin. In laminae III and IV, although there was no correlation between neurokinin-1 cell activation and stimulus intensity, the total Fos count in these layers was inversely related to the depth of cooling. Thus, neurons in laminae III and IV may also play a role in thermoreception. PMID- 10366029 TI - Increased calpain I-mediated proteolysis, and preferential loss of dephosphorylated NF200, following traumatic spinal cord injury. AB - We investigated the hypothesis that the Ca2+-activated protease calpain is involved in the pathophysiology of spinal cord injury, and is linked to the proteolytic degradation of cytoskeletal proteins. We report here that levels of calpain I (mu-calpain)-mediated spectrin breakdown products are increased by 15 min post-injury, with peak levels reached by 2 h post-injury. The dephosphorylated form of the neurofilament protein NF200 is substantially lost over the same time-period. A 35-g compressive injury was applied to the midthoracic rat spinal cord for 1 min, and animals were killed at 15 min, 1, 2, 4, 8, 16, and 24 h post-injury. Calpain I-mediated spectrin breakdown products accumulated post-injury, with peak levels reached at 2 h. Secondly, we have demonstrated a progressive loss of the 200,000 mol. wt neurofilament protein NF200, a cytoskeletal calpain substrate, which began within 1-2 h post-injury. Densitometric analyses confirmed that loss of NF200 is a substrate-specific phenomenon, since (i) dephosphorylated NF200 was preferentially lost while phosphorylated NF200 was relatively spared, and (ii) actin, which is not a substrate for calpain, was relatively spared following spinal cord injury. Finally, we demonstrated calpain I-mediated spectrin breakdown within NF200 positive neuronal processes post-injury. We conclude that the accumulation of spectrin breakdown products is temporally and spatially correlated with loss of dephosphorylated NF200 after spinal cord injury. PMID- 10366030 TI - The electroreceptor organ of the catfish, Ictalurus melas, as a model for cisplatin-induced ototoxicity. AB - The ototoxic side-effects of the anti-cancer drug cisplatin (cis diaminedichloroplatinum) have been widely investigated. However, the exact site of action remains unclear. In this study, the electroreceptor organ of the freshwater catfish Ictalurus melas is used as a model for examining the acute effects of cisplatin. The sensory cells in the electroreceptor organ are homologous to the inner hair cells in the cochlea of mammals. The effects of cisplatin administration can be investigated by in vivo recording of the spike trains from the electroreceptor organ primary afferents. Exposure of electroreceptor organs to 330 microM cisplatin for 1 h causes the spontaneous activity to drop, the overall sensitivity to diminish and the shape of the frequency characteristics to change. These effects persist in the week after administration. Control levels have returned at day 22. These results demonstrate an acute and, with considerable hysteresis, reversible cisplatin effect on the electroreceptor organs, which is to a large extent consistent with the cisplatin induced effects in isolated hair cells in mammals. The time-course of the effect supports the hypotheses that ion channels are blocked immediately by cisplatin administration, and that cisplatin metabolites disturb enzymatic cellular processes. PMID- 10366031 TI - Effects of choline and other nicotinic agonists on the tectum of juvenile and adult Xenopus frogs: a patch-clamp study. AB - We have used anatomical methods and whole-cell patch-clamp recording to assess the distribution of nicotinic receptors in the tectum of Xenopus frogs and to measure effects of nicotinic ligands (carbachol, cytisine and nicotine) on glutamatergic spontaneous miniature excitatory postsynaptic currents. Our results confirm that retinotectal axons account for the majority of nicotinic receptors in the tectum and that nicotinic agonists exert presynaptic effects that increase the rate of transmitter release on to tectal cells. The nicotinic blockers mecamylamine and methyllycaconitine reduced responses to carbachol and cytisine. A small percentage of cells also showed postsynaptic responses. We have assessed whether there are developmental changes in the frequency of occurrence of spontaneous miniature excitatory postsynaptic currents. The first three months post-metamorphosis fall within the critical period for the dramatic plasticity displayed by binocular inputs during development in Xenopus. During this period, visual activity governs the formation of orderly maps relayed from the ipsilateral eye via the cholinergic projection from the nucleus isthmi to the tectum. In this study, we have found that critical-period tecta (two to 12 weeks postmetamorphosis) tend to have higher spontaneous activity than do older tecta (two to 69 weeks postmetamorphosis), and that nicotinic agonists increase that activity in both groups, with the result that the peak rates in response to nicotinic agonists are higher during the critical period than later. We also investigated the possible role of choline as an agonist of nicotinic receptors in the tectum. We have found that choline, as well as carbachol and cytisine, can cause a reversible increase in the rate of miniature excitatory postsynaptic currents. This result may help to explain how the isthmotectal projection, which accounts for the overwhelming majority of cholinergic input to the tectum, can exert effects on retinotectal terminals even though there are no morphologically identifiable synapses between the two populations. We have examined the morphology of cells filled with biocytin during the patch-clamp experiments, and we find that cells with dendrites in the stratum zonale, a layer with particularly dense input from the contralateral nucleus isthmi, have higher spontaneous activity than cells with dendrites that do not extend into that layer. Nicotinic agonists increased the activity recorded in both classes of cells. In addition, four pretectal cells were identified. Nicotinic agonists increased the rate of spontaneous activity recorded in that population. The results indicate that retinotectal transmission in the superior colliculus can be increased presynaptically by activity of the cholinergic projections of the nucleus isthmi. This modulation may be the basis for observations that blocking of cholinergic input disrupts the formation of topographic retinotectal projections. Moreover, the ability of choline to activate these receptors suggests that this metabolite of acetylcholine may permit paracrine activation of presynaptic receptors even though the tectum contains high acetylcholinesterase activity. PMID- 10366032 TI - Cyclic AMP responsive element binding protein phosphorylation and DNA binding is decreased by chronic lithium but not valproate treatment of SH-SY5Y neuroblastoma cells. AB - Mood stabilizing drugs decrease central nervous system cyclic AMP signaling. We report here that chronic, but not acute treatment with lithium chloride in human neuroblastoma SH-SY5Y cells, inhibits phosphorylation of cyclic AMP responsive element binding protein and cyclic AMP responsive element DNA binding induced by the adenylyl cyclase activator forskolin, but has no effect on constitutive expression of cyclic AMP responsive element binding protein. These results are consistent with an effect of lithium to blunt the cyclic AMP signal transduction pathway. Such an effect is not shared by the other commonly prescribed mood stabilizer, sodium valproate. Our results suggest that cyclic AMP responsive element binding protein regulated gene expression may be relevant to the long term prophylactic effect of lithium. Furthermore, sodium valproate, which is also effective in bipolar disorder, would appear to act on other pathways to bring about its therapeutic effects. PMID- 10366033 TI - Analysis of potentiation in the cerebral ganglion of Aplysia. AB - Quantal analysis was used to characterize synaptic transmission between A and B neurons in the cerebral ganglion of Aplysia in control and during slow developing potentiation, a form of synaptic plasticity exhibited by these synapses. Control values of mean quantal content (m) and quantal size (q) estimated by the method of coefficient of variation (CV) were m approximately 6, q approximately 56 microV in the solution with Ca2+/Mg2+ = 5/200 and m approximately 18, q approximately 41 microV in the solution with Ca2+/Mg2+ = 55/150. There was a good correlation between an increase in the amplitude of excitatory synaptic potential and an increase in calculated quantal content (m(cv)) during potentiation. A decrease of Ca2+/Mg2+ ratio in the bath solution allowed observation of transmission failures and in some cases regular peaks on excitatory postsynaptic potential amplitude histograms. The latter provided more direct estimate of the quantal size. Induction of the potentiation in this solution, however, became difficult. In cases of successful potentiation induction, probability of failures was less than in control; distances between histogram peaks, reflecting quantal size remained the same. The results obtained in this study support a hypothesis that potentiation of the synaptic transmission between A and B neurons of Aplysia is primarily due to an increase of transmitter release. PMID- 10366034 TI - Astrocytes, oligodendroglia, extracellular space volume and geometry in rat fetal brain grafts. AB - Fetal neocortex or tectum transplanted to the midbrain or cortex of newborn rats develops various degrees of gliosis, i.e. increased numbers of hypertrophied, glial fibrillary acidic protein-positive astrocytes. In addition, there were patches or bundles of myelinated fibres positive for the oligodendrocyte and central myelin marker Rip, and increased levels of extracellular matrix molecules. Three diffusion parameters--extracellular space volume fraction alpha (alpha = extracellular volume/total tissue volume), tortuosity lambda (lambda = square root(D/ADC), where D is the free and ADC is the apparent tetramethylammonium diffusion coefficient) and non-specific uptake k'--were determined in vivo from extracellular concentration-time profiles of tetramethylammonium. Grafts were subsequently processed immunohistochemically to compare diffusion measurements with graft morphology. Comparisons were made between the diffusion parameters of host cortex and corpus callosum, fetal cortical or tectal tissue transplanted to host midbrain ("C- and T-grafts") and fetal cortical tissue transplanted to host cortex ("cortex-to-cortex" or C-C grafts). In host cortex, alpha ranged from 0.20 +/- 0.01 (layer V) to 0.21 +/- 0.01 (layers III, IV and VI) and lambda from 1.59 +/- 0.03 (layer VI) to 1.64 +/- 0.02 (layer III) (mean +/- S.E.M., n = 15). Much higher values were found in "young" C-grafts (81-150 days post-transplantation), where alpha = 0.34 +/- 0.01 and lambda = 1.78 +/- 0.03 (n = 13), as well as in T-grafts, where alpha = 0.29 +/- 0.02 and lambda = 1.85 +/- 0.04 (n = 7). Further analysis revealed that diffusion in grafts was anisotropic and more hindered than in host cortex. The heterogeneity of diffusion parameters correlated with the structural heterogeneity of the neuropil, with the highest values of alpha in gray matter and the highest values of lambda in white matter bundles. Compared to "young" C grafts, in "old" C-grafts (one year post-transplantation) both alpha and lambda were significantly lower, and there was a clear decrease in glial fibrillary acidic protein immunoreactivity throughout the grafted tissue. In C-C-grafts, alpha and lambda varied with the degree of graft incorporation into host tissue, but on average they were significantly lower (alpha = 0.24 +/- 0.01 and lambda = 1.66 +/- 0.02, n = 8) than in young C- and T-grafts. Well-incorporated grafts revealed less astrogliosis, and alpha and lambda values were not significantly higher than those in normal host cortex. The observed changes in extracellular space diffusion parameters could affect the movement and accumulation of neuroactive substances and thus impact upon neuron-glia communication, synaptic and extrasynaptic transmission in the grafts. The potential relevance of these observations to human neuropathological conditions associated with acute or chronic astrogliosis is considered. PMID- 10366035 TI - Immunosenescence: impact in the young as well as the old? PMID- 10366036 TI - DNA damage in lymphocytes of elderly patients in relation with total antioxidant levels. AB - DNA damage may occur as a result of an imbalance between the production and removal of free radicals, a process in which age plays an outstanding role. The purpose of this study was to analyze the relationship between total antioxidants and DNA damage in a sample of old age people in Mexico City. The sample included a total of 88 subjects, 15 males and 69 females, with a mean age of 65.5 years old (range between 60 and 79 years old), all of whom had lived in Mexico City during the last 10 years and had been diagnosed as clinically healthy. Results showed that 52% of the subjects presented DNA damage in peripheral blood lymphocytes which was assessed through an alkaline unicellular electrophoresis procedure (Comet Test), regardless of total antioxidant serum levels quantified through a colorimetric method (Randox Kit). Higher non-damage occurrences were observed in subjects with low antioxidant levels, a difference that was statistically significant (P < 0.05). Furthermore, the highest incidence of damaged cells was observed in subjects belonging to the 70-years-old-and-above group (P < 0.05). As to the magnitude and intensity of the damage associated to total antioxidant concentrations, a trend toward greater DNA damage in subjects with low serum levels was observed. It is concluded that low antioxidant levels are not always indicative of oxidative strain and therefore should not be considered as predictors of DNA damage in this population. PMID- 10366037 TI - Granulocyte and natural killer activity in the elderly. AB - The deterioration of the immune system in ageing, 'immunosenescence', is thought to contribute to increased morbidity and mortality from infections and possibly autoimmune diseases and cancer. The most profound changes involve effector and immunoregulatory T-cell functions. Immunosenescence appears also to be related to changes in non specific immunity as well. In the present study we have assessed superoxide production, chemotaxis and the expression of the apoptosis-related molecule APO1/Fas (CD95) on neutrophils (PMN) from young and old subjects. Furthermore, we have measured the basal natural killer (NK) activity of young and elderly subjects and we have compared the number of CD16+ cells found in these two groups. We observed a significant decrease age-related both of formation of O2- and chemotaxis whereas no significant correlation between age and the expression of CD95 on granulocyte membrane was demonstrated, suggesting that an increase age-related of CD95-linked apoptosis of PMN should be not an important determinant in the decreased PMN function. We also observed a significant correlation between age and NK activity. The decreased NK cell function was not due to a decreased number of NK cells in effector cell preparations since the number of CD16+ cells was significantly increased in old subjects. In conclusion, our results show that in the elderly there is also a deficit of the aspecific immunity that might play a role in the pathogenic mechanisms of the immunosenescence. PMID- 10366038 TI - Ageing and endocrine cells of human duodenum. AB - Motility and secretory disorders of the gastrointestinal tract and associated glands increase with ageing. The duodenum contains several peptide/amine producing cells that play an important role in regulating gastrointestinal motility and secretion. The present study was performed to elucidate changes in these cells that may have arisen as a result of ageing. A total of four age groups of subjects, aged 1-2, 20-29, 40-49 and 60-69 years were studied. The various endocrine cell types were identified by immunohistochemistry and quantified by computerized image analysis, and two parameters were determined; the number of cells/mm3 epithelial cells and the cell secretory index (CSI), which indicates the immunoreactive secretory granule content of the endocrine cells. Chromogranin A- and serotonin-immunoreactive (IR) cells were fewer in 1-2 year-olds than in 20-29-year-olds. Gastrin/CCK-IR cells were significantly more numerous in 1-2-year-olds and 60-69 years-olds than in 20-29-year-olds. Somatostatin-IR cells were more numerous in the 40-49-year-olds than in the 20-29 years-olds. The CSI was higher in chromogranin A-, gastric inhibitory polypeptide (GIP)-, somatostatin- and gastrin/CCK-IR cells in 1-2-year-olds than in 20-29 year-olds. There was no significant sex difference regarding the numbers and CSI of other endocrine cell types. This study established the absence of sex-related differences in all endocrine cell types investigated, regarding numbers and physiological activity. Age, on the other hand, was shown to be associated with changes in the numbers of CCK-, somatostatin- and serotonin-IR, which may have some bearing on the gastrointestinal disorders of the elderly. PMID- 10366039 TI - Age-related changes in the NADPH-diaphorase-positive neuronal perikarya of the dorsolateral column of the periaqueductal gray in the rat. AB - The distribution of reduced nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) positive neuronal perikarya in the rostral, middle and caudal parts of the dorsolateral periaqueductal gray (DLPAG) in 3-, 12- and 26 month-old rats was compared by means of histochemistry and computer assisted image analysis. The total number, the maximum diameter, the cross-sectional area and the optical density (OD) of the NADPH-d positive neurons were analyzed. The results demonstrate that there are no significant differences in any of the investigated parameters between the left and the right parts of the same age and of the same level. The total cell number in the DLPAG of 26-month-old rats was significantly decreased in comparison with 3- and 12-month-old rats. The cross sectional area increased between 3 and 12 months of age, and decreased in 26 month-old rats in comparison with 12-month-old rats. The OD increased significantly between 3 and 12 months of age, and decreased significantly between 12 and 26 months of age only at caudal level. The observed mild changes could lead to alterations in the brain physiology of aging. PMID- 10366040 TI - Adaptations in lactate dehydrogenase and its isozymes in aging mammalian myocardium: interaction of exercise and temperature. AB - The responses of the left and right ventricles (LV and RV) to physical conditioning in cold (25 degrees C) and thermoneutral temperatures (35 degrees C), with special reference to lactate dehydrogenase (LDH) and its isoenzyme profile, were studied in the 2-month (young)- and 12-month (middle-aged)-old rats. Moderate hypertrophy was a common observation irrespective of age, region and swim temperature. LV, however, hypertrophied to a significantly lesser extent in the middle-aged, than the RV. Blood Lactate (La) content showed a decline in the trained rather than their untrained counterparts. LDH activity decreased with age. Swim training induced elevations in the enzyme activity. The isoenzyme profile was suitably and efficiently altered in the LV and RV of trained animals to meet the arising O2 demands. The above adaptations were best seen in the young and in the animals trained at thermoneutral temperatures. Thus it is suggested that young age is very apt for initiation of training programs although middle age is not so late. Swimming in water near body temperature is emphasised as a more preferred environment to cold water, in order to derive maximal exercise associated beneficial effects. PMID- 10366041 TI - Evidence that FGF receptor signaling is necessary for endoderm-regulated development of precardiac mesoderm. AB - Endoderm cells in the heart forming region (HFR endoderm) of stage 6 chicken embryos are required to support the proliferation and terminal differentiation of precardiac mesoderm cells in vitro. The endoderm's effect can be substituted by growth factors, including members of the fibroblast growth factor (FGF) family. However, direct implication of FGFs in this process requires evidence that inhibition of FGF signaling interferes with proliferation and/or terminal differentiation. This report examines the consequences of treating endoderm/precardiac mesoderm co-explants with agents that inactivate FGF receptors. Using sodium chlorate, which prevents FGF ligand-receptor interaction, it was observed that the percentage of S-phase precardiac mesoderm cells was markedly reduced, suggesting that cell proliferation was inhibited. To more specifically affect FGF signaling, the explants were treated with an antibody that recognizes an extracellular domain of FGF receptor-1 (FGFR-1). This treatment similarly inhibited cell proliferation. Although both agents modestly delayed cardiac myocyte differentiation as indicated by the contractile function, expression of alpha-sarcomeric actin was not affected. These findings provide additional evidence that an intact FGF signaling pathway is required during heart development. PMID- 10366042 TI - Expression of the c-fos gene induced by parathyroid hormone in the bones of SAMP6 mice, a murine model for senile osteoporosis. AB - SAMP6 mice are a murine model for senile osteoporosis, characterized by low peak bone mass seen at 4 or 5 months of age. Parathyroid hormone (PTH)-induced c-fos expression was examined in the bones, bone-marrow cells and kidney tissues of 2 month-old male SAMP6 mice. SAMP2 mice, which have a higher peak bone mass, were used as controls. The expression of c-fos in the bone peaked at 30 min after 60 microg/kg of human PTH(1-34) administration. After peaking, the expression fell quickly in SAMP2 mice. This decrease in expression was delayed in SAMP6 mice and the expression was higher at 1 h than in SAMP2 mice. The phenomenon observed in the bone appears to be tissue specific as it was not seen in the bone-marrow cells or kidney tissue. Immunohistochemical studies showed that c-Fos protein was localized to the nuclei of some of the osteocytes and a few of the osteoblasts in the cortical bone, and that osteocytes expressing c-Fos protein increased after PTH treatment. These results suggest that osteocytes might contribute to the maintenance of higher levels of c-fos expression in the bones of SAMP6 mice and may be related to cortical osteopenia in these mice by modulating bone remodeling and/or modeling. PMID- 10366043 TI - Maternal factors associated with severity of birth defects. AB - OBJECTIVE: Due to inbreeding and nutritional factors, the Bedouin Arabs represent a high risk population for birth defects. The severity of birth defects is probably related to the time and extent of interference with embryogenesis. The present study was aimed at identifying factors associated with severity of birth defects, in pregnancies of Bedouin women examined at a third level ultrasound clinic. METHODS: The study population consisted of 295 Bedouin women who attended an ultrasound clinic at the Soroka Medical Center between 1990 and 1996. The case group included 188 women carrying fetuses with severe birth defects, defined as incompatible with life or which significantly interfere with normal living. For those defects the option of pregnancy termination was discussed. The comparison group consisted of 107 women whose fetuses were diagnosed with mild defects. RESULTS: Women carrying fetuses with severe birth defects had more pregnancies and more deliveries than women carrying fetuses with mild defects (P = 0.005, P = 0.04, respectively). The severity of defects was found to be unrelated to maternal age, consanguinity, residence, birth order, previously uncompleted pregnancies and birth defects in the family. CONCLUSIONS: Higher birth order was associated with severity of birth defects detected at the second trimester. PMID- 10366044 TI - Relationship of gestational age and cervical dilation to the timing of delivery. AB - OBJECTIVE: To determine the effect of gestational age and cervical dilation on pregnancy continuation in women with idiopathic preterm labor who were treated with parenteral tocolysis. METHOD: A total of 950 women with singleton gestations, intact membranes and preterm labor treated with tocolysis prior to 34 weeks' gestation were retrospectively studied. These women were identified from the March of Dimes prematurity prevention program database. For analysis, women were categorized into five gestational age groups and three cervical dilation groups. The primary outcomes measured were the percentage of women who remained undelivered at 48 h and at 14 days post-initiation of therapy. RESULT: Overall, 82% of women remained undelivered after 48 h and 65% remained undelivered at 14 days. As cervical dilation advanced, the number of women remaining undelivered at 48 h and 14 days significantly decreased. However, even at > or = 4 cm, 52% of women remained undelivered at 48 h. If the cervix was dilated < 2 cm, gestational age did not influence the number of days gained prior to delivery. However, if the cervix was dilated > or = 2 cm, women at < 25 weeks' gestation were more likely to deliver compared to women at the same dilation but with more advanced gestational ages. CONCLUSION: Overall, 82% of women in preterm labor and 52% of those presenting with > or = 4-cm cervical dilation, delivered after 48 h. Therefore there appears to be ample opportunity for most women in preterm labor with intact membranes, even those at advanced dilations, to receive a complete course of corticosteroid therapy. PMID- 10366045 TI - Pregnancy following cardiac valve replacement surgery. AB - OBJECTIVES: To study the outcome of pregnancy in women with artificial heart valves and to compare the maternal and perinatal outcome in mechanical and bioprosthetic valves. METHOD: Retrospective analysis of 34 pregnancies in 29 women who conceived after cardiac valve replacement was carried out. RESULTS: The majority of women (76.4%) delivered within 5 years of valve replacement. Anticoagulants were administered in 79.4% of pregnancies. Maternal mortality was 2.9% and maternal morbidity in the form of heart failure, atrial fibrillation, valve thrombosis, thromboembolism, bleeding complications and non-functioning prostheses were 2.9%, 5.8%, 2.9%, 2.9%, 11.7% and 2.9%, respectively. The incidence of prematurity was 5.8% and small for gestational age babies was 11.7%. There was no case of abortion. Two babies (5.8%) were still born, one of which had malformations. Maternal complications were significantly higher in women with bioprostheses, though the complications were more grave in the mechanical prostheses group. The perinatal outcome was almost similar in both the groups. CONCLUSION: The perinatal outcome was not different in women with bioprosthetic valves from the ones with mechanical prostheses, but the maternal morbidity was more in women with bioprosthetic valves. Coumarin derivatives were safe and effective and did not lead to embryopathy. PMID- 10366046 TI - Routine heart and lung auscultation in prenatal care. AB - OBJECTIVE: To determine the value of routine heart and lung auscultation (HLA) in pregnant women. METHODS: Descriptive study by record review of maternal case files. RESULTS: Of 3191 mothers, there were 24 with heart disease (0.8%) and 53 with lung disease (1.7%). On routine HLA, only two new cases of heart disease, both with asymptomatic valvular conditions, were detected, and none of lung disease. CONCLUSION: Routine chest auscultation is of very little value in prenatal care. PMID- 10366047 TI - Tetanus toxoid and congenital abnormalities. AB - OBJECTIVE: To study the human teratogenic potential of tetanus vaccination during pregnancy. METHODS: Pair analysis of cases with congenital abnormalities and matched healthy controls was performed in the large population-based dataset of the Hungarian Case-Control Surveillance of Congenital Abnormalities, 1980-1994. RESULTS: Of 35 727 pregnant women who had babies without any defects in the study period (control group), 33 (0.09%) were vaccinated with tetanus. Of 21563 pregnant women who had offspring with congenital abnormalities, 25 (0.12%) had tetanus vaccination. This difference was not significant (P = 0.39). The case control pair analysis confirmed the safety of tetanus vaccination during pregnancy, particularly in the second and third months of gestation, i.e. during the critical period for congenital abnormalities. CONCLUSION: Tetanus vaccination during pregnancy appears not be teratogenic to the fetus. Thus, there is no contraindication, if the use of tetanus toxoid is necessary during pregnancy. PMID- 10366048 TI - Human papillomavirus with co-existing vulvar vestibulitis syndrome and vestibular papillomatosis. AB - OBJECTIVE: The role of HPV infection in cases of vulvar papillomatosis and vulvar vestibulitis syndrome is still unclear and data from the literature is controversial. In this study we intended to investigate the prevalence of viral infection, with a multidisciplinary approach, in cases with a co-existence of the two patterns. METHOD: Sixteen consecutive cases with diagnosis of vulvar vestibulitis syndrome and co-existence of vestibular papillomatosis were enrolled in the study and investigated by the means of vulvar cytology, vulvoscopy, histology, ViraPap and Polymerase Chain Reaction. RESULT: Cytology, vulvoscopy and histology did not demonstrate suitable accuracy for the diagnosis. Viral DNA identification revealed two (12.50%) positive cases using PCR and one (6.25%) positive case with ViraPap. CONCLUSION: The results of the present investigation indicate that even in cases of co-existing vulvar papillomatosis and severe vulvar vestibulitis syndrome, the prevalence of HPV infection is too low to be considered causal. PMID- 10366049 TI - Invasive carcinoma of the uterine cervix in Iran. AB - OBJECTIVE: To review diagnostic aspects, treatment measures and pathologic findings in patients diagnosed and treated for invasive cervical carcinoma in Shiraz University of Medical Sciences hospitals. METHOD: The hospital records of all patients admitted to the Shiraz University of Medical Sciences affiliated hospitals (Nemazee and Saadi hospitals) with newly diagnosed carcinoma of the uterine cervix between January 1985 and December 1995, were reviewed. RESULTS: 204 newly diagnosed patients were admitted with invasive cervical carcinoma of which 61 (30%) were in stage I (FIGO), 96 (47%) in stage II, 34 (16.6%) in stage III and 13 (6%) in stage IV of the disease. Of these 204, the given treatments included surgery in 88 patients, internal and external pelvic radiotherapy in 141, both hysterectomy and pelvic radiotherapy in 22, chemotherapy in eight and cryotherapy in one patient. We found significantly positive relations with stage of the disease and chest roentgenogram (P < 0.02), intravenous pyelogram (P < 0.0002), sigmoidoscopy (P < 0.006) and cystoscopy (P < 0.0045). Of the total number of pathologic specimens, squamous cell carcinoma was reported in 88% and adenocarcinoma in 11%. Recurrence rates of up to 15.5% for stage II and 5% for stage I were calculated. Among the complications for which hospital admission was required, genito-urinary complications were the most frequent. CONCLUSION: The recurrence rate in stage I is lower than the estimated recurrence risk in several other studies which can be due to a real advantage of the center's practice. This may pose the question of indispensability for the advanced and expensive diagnostic paraclinical work-ups. PMID- 10366050 TI - Monitoring parameters in the management of patients with tubo-ovarian complexes. AB - OBJECTIVE: The aim of the study was to determine which of the monitored parameters is most suitable for following the treatment of tubo-ovarian complex (TOC). METHODS: In 35 patients treated conservatively for TOC, serum levels of C reactive protein (CRP), erytrocyte sedimentation rate (ESR) and tumor-associated antigen 125 (CA-125) were determined at the beginning as well as on days 5, 10, 15 and 20 of treatment. TOC size was measured ultrasonographically prior to and on day 20 of treatment. We compared the dynamics of these parameters. Spearman's correlation test was used to establish the connection between levels of CRP, ESR, CA-125 and the size of the TOC. RESULTS: CRP follows the course of acute inflammation quickly, ESR and CA-125 with some delay. The correlation between the size of the tumor on admission and the CRP levels was established on days 5-10, ESR from the beginning until day 15 and the CA-125 levels on days 10-20 of treatment. On day 20, only the levels of CA-125 correlated with the size of the TOC measured on the same day (R = 0.49, P = 0.01) and only the difference between its levels correlated with the difference in TOC (R = 0.41, P = 0.01). CONCLUSION: Determination of CRP, and somewhat less ESR, is more suitable for following the treatment of TOC in the acute phase while CA-125 and measuring of TOC size is more suitable in the subacute phase. PMID- 10366051 TI - Efficacy, tolerability, and rare side effects of tibolone treatment in postmenopausal women. AB - OBJECTIVE: The purpose of this study was to assess the tolerability and side effects of tibolone (Livial, Organon), a synthetic steroid analogue for the treatment of postmenopausal symptoms, in a large population of patients. METHOD: 1189 postmenopausal patients were included in this study. The patients' blood pressure, body weight, general complaints, and the severity of their climacteric complaints were documented at baseline and after 4 months of tibolone treatment. RESULTS: Tibolone significantly relieved all of the classical menopausal complaints. The proportion of patients with bleeding problems dropped significantly from 15.9% to 6.8%. Other complaints, such as headache, vertigo, nervousness, breast tenderness, and hirsutism were also significantly less frequent than before treatment. Only few women reported other rare side effects, and only 14.4% of women discontinued treatment prematurely. CONCLUSION: Tibolone provides an efficient and safe means of treating the postmenopausal syndrome in every-day practice. PMID- 10366052 TI - Confidential enquiry of stillbirths in current obstetric practice. AB - OBJECTIVE: Confidential panel enquiry into sub-optimal factors relating to stillbirths. METHOD: All 121 stillbirths in KK Women's and Children's Hospital in the years 1995 and 1996 were studied. Three assessors reviewed the case records of each death, and panel consensus was reached regarding sub-optimal antenatal care and factors leading to stillbirths. RESULT: The incidence rate of stillbirth was 4.04 per 1000 deliveries. A total of 76 cases (62.8%) were found to have grade II and III sub-optimal factors in their management. Patients themselves were involved in the sub-optimal management of their own pregnancy in 52.9% of the stillbirths. Primary healthcare givers were involved in 8.3% of all stillbirths, specialist caregivers 12.4% and antenatal care system 4.1%. CONCLUSION: The study has identified sub-optimal antenatal management in over 60% of cases. As patients' factors form the major contribution towards sub-optimal care, management strategy aimed towards improving patients' education and compliance to antenatal care should be a priority. PMID- 10366053 TI - Swiss consensus guidelines for hysterectomy. Swiss Society of Gynecology and Obstetrics, Switzerland. AB - OBJECTIVE: The quality of the indication for hysterectomy is widely discussed at present. In early 1996, the committee for quality assurance of the Swiss Society of Gynecology and Obstetrics decided to set up nationally accepted guidelines for the indication of hysterectomy. METHODS: A modified Delphi approach was used. In a first step, general guidelines and actions prior to hysterectomy were defined. An expert panel of 17 Swiss gynecologists rated 74 frequent indications, twice for appropriateness (more benefits than risks for the patient), once for necessity (n = 34; procedure has to be offered or discussed with the patient), and outlined suggestions to be performed prior to hysterectomy. RESULTS: In a home rating round before the first panel met, there was an agreement rate of 48%. In 45% we observed neither agreement nor disagreement; in 7% we found disagreement. After the panel discussion 89% of experts agreed, 11% were indeterminate, and there was no disagreement. The necessity ratings showed agreement in 68% while 32% were indeterminate. The average median rating on a 1-9 point scale (1 = extremely inappropriate, 9 = extremely appropriate or necessary) was 5.4 over all single indications for appropriateness and 7.8 in single indications for necessity. After a second panel for consensus all panelists agreed on both appropriateness and necessity. CONCLUSION: The results of the appropriateness and necessity consensus presented in this paper reflect the findings of a 17 member Swiss panel. This joint effort by a medical society may be a step towards the direction of a peer controlled healthcare system. PMID- 10366054 TI - The outcome of the first VBAC program in Thailand. PMID- 10366055 TI - Hydatidiform mole in Okinawa islands and mainland Japan. PMID- 10366056 TI - Intrapartum rupture of the uterus--19 years' experience. PMID- 10366057 TI - Vesico-vaginal fistula in Benin City, Nigeria. PMID- 10366058 TI - Value of postoperative treatment after cutting of a broad base intrauterine septum. PMID- 10366059 TI - FIGO Committee Report. FIGO Committee for the Ethical Aspects of Human Reproduction and Women's Health. International Federation of Gynecology and Obstetrics. PMID- 10366060 TI - ACOG educational bulletin. Special problems of multiple gestation. Number 253, November 1998 (Replaces Number 131, August 1989). American College of Obstetricians and Gynecologists. AB - The incidence of twins, triplets, and higher-order multiple gestations has increased dramatically because of widespread use of ovulation-inducing drugs and advanced assisted reproductive techniques. There is considerable perinatal/maternal morbidity and mortality associated with multifetal gestations. The practicing obstetrician managing these high-risk patients should be familiar with their special antepartum and intrapartum problems. The obstetrician gynecologist unaccustomed to caring for patients with a multifetal gestation should consult with maternal-fetal medicine specialists who have expertise in managing these pregnancies. Better use of infertility modalities, early diagnosis of the multiple pregnancy, prevention of preterm birth, close fetal surveillance, and atraumatic labor and delivery can improve perinatal outcome in the multifetal gestation. PMID- 10366061 TI - ACOG committee opinion. Antenatal corticosteroid therapy for fetal maturation. Number 210, October 1998 (Replaces Number 147, December 1994). Committee on Obstetric Practice. American College of Obstetricians and Gynecologists. PMID- 10366062 TI - Obesity and elevated intraocular pressure. PMID- 10366063 TI - Diabetes and visual loss. PMID- 10366064 TI - Immediate argon laser peripheral iridoplasty for acute attack of PACG (addendum to previous report) PMID- 10366065 TI - Success of cataract surgery in diabetics. PMID- 10366066 TI - Dry eye syndrome. PMID- 10366067 TI - Dorzolamide-timolol combination versus concomitant administration of its components. PMID- 10366068 TI - Botulinum toxin therapy in exotropia. PMID- 10366069 TI - Neuro-ophthalmic manifestations of diabetes. PMID- 10366070 TI - Racial differences in the prevalence of age-related macular degeneration: the Baltimore Eye Survey. AB - OBJECTIVE: To determine the prevalence of age-related macular degeneration (AMD) and signs of age-related maculopathy in a population-based sample of blacks and whites 40 years of age or older from East Baltimore. DESIGN: Cross-sectional population-based study. PARTICIPANTS: A total of 5308 black and white subjects received a screening eye examination that included fundus photography. MAIN OUTCOME MEASURES: Stereoscopic color fundus photographs were graded for the presence and severity of drusen, pigmentary abnormalities, geographic atrophy, and choroidal neovascularization in the macula. RESULTS: Drusen > or = 64 microm were identified in about 20% of individuals in both groups, but large drusen (>125 microm) were more common among older whites (15% for whites versus 9% for blacks over 70). Pigmentary abnormalities were also more common among older whites (7.9% for whites versus 0.4% for blacks over 70). Age-related macular degeneration was more prevalent among whites than blacks. The prevalence of AMD was 2.1 % among whites over 70 years of age. No cases of AMD were detected among 243 black subjects in this age group. Logistic regression adjusting for age, sex and smoking (current, former, or never) detected an odds ratio of 4.4 (95% confidence interval: 1.5-12.4) for whites with AMD compared with blacks. CONCLUSION: Although drusen are common in both blacks and whites over the age of 40, more severe forms of age-related maculopathy and late AMD are more prevalent in older whites. PMID- 10366071 TI - Age-related maculopathy in a multiracial United States population: the National Health and Nutrition Examination Survey III. AB - OBJECTIVE: To investigate the prevalence of and risk factors for age-related maculopathy (ARM) in three racial/ethnic groups: non-Hispanic whites, non Hispanic blacks, and Mexican-Americans. DESIGN: A nationally representative population-based, cross-sectional study. PARTICIPANTS: A total of 8270 persons 40 years of age or older, a sample of the Third National Health and Nutrition Examination Survey. MAIN OUTCOME MEASURES: Age-related maculopathy was determined by the grading of fundus photographs using a standardized protocol. RESULTS: The prevalence of any ARM in the civilian noninstitutionalized United States population including those 40 years of age or older was 9.4% (95% confidence interval [CI], 8.2, 10.6) as estimated from the sample. After adjusting for age, there was no difference in the prevalence of early ARM (defined largely by the presence of soft drusen) by ethnic/racial group. However, for the less frequent component lesions of early ARM (increased retinal pigment and retinal pigment epithelial depigmentation), the odds ratios (95% CIs) comparing non-Hispanic blacks to non-Hispanic whites were 0.47 (0.31, 0.74) and 0.59 (0.33, 1.04), respectively, and for comparing Mexican-Americans to non-Hispanic whites, they were 0.41 (0.21, 0.81) and 0.72 (0.44, 1.19), respectively. For late ARM, the odds ratio (95% CI) for non-Hispanic blacks compared to non-Hispanic whites was 0.34 (0.10, 1.18) and for Mexican-Americans compared to non-Hispanic whites, it was 0.25 (0.07, 0.90). Other than age, none of the personal, medical, or physiologic variables studied were statistically significantly associated with any of the ARM endpoints in any of the three races/ethnic groups. CONCLUSION: Overall, rates of any ARM (including all early and late lesions) are not significantly different among non-Hispanic blacks, Mexican-Americans, and non Hispanic whites. However, the rates of individual lesions suggest that non Hispanic whites and Mexican-Americans may be protected against retinal pigment abnormalities and lesions associated with late ARM. There appears to be little influence of personal, medical, and environmental factors studied on these results. Further studies in larger populations of older persons in these ethnic groups would likely clarify these relations. PMID- 10366072 TI - Refractive errors in an older population: the Blue Mountains Eye Study. AB - OBJECTIVE: To determine prevalence and associations with refractive errors in a defined older population. DESIGN: Cross-sectional study. PARTICIPANTS: A total of 3654 residents, aged 49-97, of the Blue Mountains, west of Sydney, Australia. METHODS: Comprehensive questionnaire and detailed eye examination, including refraction. MAIN OUTCOME MEASURES: Refractive error of phakic eyes, age, gender, and education. RESULTS: Prevalence rates were determined for myopia (15%), hyperopia (57%), and emmetropia (28%). Hyperopia prevalence was age-related, increasing from 36% in persons aged <60 years to 71 % of persons aged > or = 80 (P < 0.0001), whereas myopia prevalence decreased with age, from 21 % in persons aged <60 years to 10% of persons aged > or = 80 years (P < 0.0001). Younger myopic subjects in this population reported first wearing distance correction at a significantly younger age than older subjects, P < 0.0001. After adjustment for age, women were slightly more hyperopic (mean +0.75 diopters [D]) than men (mean +0.59 D, P = 0.0012. The gender-adjusted mean spherical error increased with age from +0.03 D in persons aged <60 years to +1.2 D in persons aged > or = 80 years (P < 0.0001). The gender-adjusted mean cylinder power also increased with age, from -0.6 D in persons aged <60 years to -1.2 D in persons aged > or = 80 years (P < 0.0001). The mean axis of astigmatism was "against the rule" in all age groups. Anisometropia increased with age, from a mean of 0.4 D in persons aged <60 to 0.9 D in persons aged > or = 80 years (P < 0.0001). Higher education was associated with myopia in men (P = 0.009) but not in women (P = 0.21) after adjustment for age. CONCLUSION: This report has documented the detailed refractive status of an older population, confirming previously described trends but also finding an apparent higher prevalence of myopia among younger members of this community. PMID- 10366073 TI - Pediatric photoscreening for strabismus and refractive errors in a high-risk population. AB - OBJECTIVE: To determine the accuracy of the MTI Photoscreener in detecting strabismus and refractive errors in children. PARTICIPANTS: One hundred children underwent MTI photoscreening followed by complete ophthalmologic examination. Six observers graded the photographs for strabismus, according to the location of the corneal light reflexes, and for refractive error, according to the size and location of the light crescent. RESULTS: The sensitivity of the MTI Photoscreener in detecting any amblyogenic factor was 80% to 91%, with a specificity of 20% to 67%. The sensitivity and specificity for particular amblyogenic factors varied widely among observers. The ranges were as follows: strabismus, sensitivity = 23% to 50%, specificity = 76% to 96%; myopia, sensitivity = 89%, specificity = 48% to 76%; hyperopia, sensitivity = 20% to 80%, specificity = 88% to 96%; and astigmatism, sensitivity = 46% to 77%, specificity = 79% to 89%. CONCLUSIONS: These results suggest caution in relying on photoscreening to detect strabismus and refractive errors in children. PMID- 10366074 TI - Vitrectomy for rhegmatogenous or tractional retinal detachment with familial exudative vitreoretinopathy. AB - OBJECTIVE: To examine the anatomic features and surgical indications of familial exudative vitreoretinopathy (FEVR) complicated with rhegmatogenous or tractional retinal detachment. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Twenty-eight eyes of 25 patients who had either clinically suspected or fully diagnosed FEVR. Of these, 25 had rhegmatogenous retinal detachment, 2 had tractional retinal detachment, and 1 had tractional retinal detachment plus vitreous hemorrhage. INTERVENTIONS: The authors carefully observed the vitreoretinal interface during surgery, studied the clinical and anatomic features of FEVR, and then evaluated the surgical results. RESULTS: The vitreoretinal adhesions were so strong in the peripheral avascular area that iatrogenic retinal breaks easily occurred in 22 of 28 eyes. In all cases, the bimanual technique with vitreous scissors and forceps was required to dissect the posterior vitreous membrane from the retinal surface. The retina was reattached in 24 of 28 cases (85.7%), and visual acuity improved in 20 eyes (71.4%). CONCLUSION: Dissection of the vitreous in the peripheral avascular area is very difficult in FEVR, and those patients for whom this procedure was not successfully performed may have a poorer prognosis. PMID- 10366075 TI - Visual outcomes after pars plana vitrectomy for epiretinal membranes associated with pars planitis. AB - OBJECTIVE: To evaluate the results of pars plana vitrectomy and membrane stripping for visually significant macular epiretinal membranes associated with chronic idiopathic pars planitis. DESIGN: Consecutive noncomparative case series. PARTICIPANTS AND METHODS: The records of all patients who underwent pars plana vitrectomy for pars planitis from 1988 through 1997 were retrospectively reviewed. Seven eyes of five patients who were diagnosed with visually significant epiretinal membranes associated with pars planitis and who underwent vitrectomy and membrane stripping were analyzed. Patients were diagnosed with pars planitis based on characteristic clinical signs and pertinent negative laboratory test results. INTERVENTION: Pars plana vitrectomy and epiretinal membrane stripping. MAIN OUTCOME MEASURES: Visual acuity and inflammatory grade were compared between the last preoperative visit and the most recent follow-up visit. Intraoperative and postoperative complications were also analyzed. RESULTS: The mean patient age was 31 years (range, 6 to 45 years). The mean duration of uveitis was 6.4 years (range, 6 months to 13 years). All patients were treated with combinations of periocular, topical, and oral corticosteroids before surgery. Five eyes had laser retinopexy, and two eyes had cryopexy to the inferior retina at the time of surgery. Five eyes had at least 3 Snellen lines of visual acuity improvement, and visual acuity in one eye worsened by 2 lines. Mean preoperative visual acuity was 20/73 (range, 20/50 to 20/300), and mean final visual acuity was 20/37 (range, 20/25 to 20/70). Five eyes had a final visual acuity of 20/40. Vitritis improved in all cases. Mean follow-up was 23 months (range, 3 to 54 months). Six of seven eyes had progressive cataract development, four of which underwent cataract extraction. No other intraoperative or postoperative complications occurred. CONCLUSIONS: Removal of epiretinal membranes associated with pars planitis can be safely performed and may result in improved visual acuity. Patients often require subsequent cataract extraction to obtain the best long-term final acuity. PMID- 10366076 TI - Chemical stability of silicone oil in the human eye after prolonged clinical use. AB - OBJECTIVE: To investigate the assumption that silicone oil is chemically stable in the human eye after prolonged clinical use as a vitreous substitute. DESIGN: Experimental study. MATERIAL: Samples of silicone oil recovered from the vitreous cavities of 25 consecutive patients up to 26 months (mean, 9.2 months) after implantation and 4 different batches of original highly purified silicone oil with a kinematic viscosity of 5000 mPa.s were analyzed. Visible silicone oil emulsification was present in 18 of the 25 eyes. INTERVENTION: Gas chromatography/mass spectroscopy (GC/MS) was used to detect and characterize low molecular weight components (LMWC). Gel permeation chromatography (GPC) was used to determine the molecular weight distribution of the silicone oils. Functional groups of the silicone oils were quantified and characterized by infrared (IR) spectroscopy and 1H nuclear magnetic resonance (1H NMR) spectroscopy. MAIN OUTCOME MEASURES: The sample oils explanted from the eyes were compared to the original oils with regard to LMWC, molecular weight distribution, and type and amount of functional groups. RESULTS: The GC/MS chromatograms showed no peaks indicative of LMWC in any of the 25 sample oils explanted from the eyes or any of the original oils. In the GPC chromatograms, the peak position occurred at the same retention times with identical signal shape in all original and sample oils, indicating that the molecular weight and the molecular weight distribution did not change after prolonged implantation. The IR spectroscopy and 1H NMR spectroscopy showed characteristic absorption bands at 1260 cm(-1) related to symmetric deformation vibration of the Si-CH3 group and at 800 cm(-1) related to the Si-(CH3)2 group. This was identical for all sample and original oils. CONCLUSIONS: Highly purified 5000-mPa.s silicone oils, used as prolonged retinal tamponades in patients with proliferative vitreoretinopathy, proliferative diabetic retinopathy, and tractional retinal detachment after central retinal vein occlusion, are chemically stable in the human eye and do not undergo chemical modification. The LMWC do not play a substantial role in the variations of oil emulsification. PMID- 10366077 TI - Pigmentary retinal dystrophy and the syndrome of maternally inherited diabetes and deafness caused by the mitochondrial DNA 3243 tRNA(Leu) A to G mutation. AB - OBJECTIVE: To study the association of retinal disease and the syndrome of maternally inherited diabetes and deafness caused by an A to G mutation in the tRNA leucine gene at base pair 3243 (A3243G) of the mitochondrial genome. DESIGN: Observational study of a genetically defined subject group. PARTICIPANTS: Thirteen subjects with the mitochondrial DNA A3243G mutation from seven different pedigrees with maternally inherited diabetes and deafness. INTERVENTION: Assessment of visual symptoms and visual acuity, dilated indirect ophthalmoscopy, retinal photography, and retinal electrophysiology. MAIN OUTCOME MEASURES: Loss of vision, funduscopic evidence of pigmentary retinal disease or diabetic retinopathy, and electrophysiologic evidence of defective functioning of the retinal pigment epithelium/photoreceptor complex. RESULTS: Funduscopic examination revealed abnormalities of retinal pigmentation in ten subjects (77%). Defects included speckled and patchy hyperpigmentation at the posterior pole of the fundus, particularly in the macular area, and varying degrees of loss of retinal pigmentation. Three subjects (23%) had visual symptoms, which included night blindness, visual loss, and photophobia. Electrophysiologic studies revealed impaired electro-oculogram responses in four of nine subjects with defects of retinal pigmentation (44%), two of whom also had much reduced scotopic and, to a lesser extent, flicker electroretinogram b wave potentials. Two subjects had diabetic retinopathy, including one with retinal depigmentation and impaired electro-oculogram activity. Both subjects with diabetic retinopathy had unilateral reduced electroretinogram responses, especially oscillatory potentials. CONCLUSIONS: Abnormalities of retinal pigmentation are common in subjects with maternally inherited diabetes and deafness caused by the mitochondrial DNA A3243G mutation. Visual symptoms, in particular loss of visual acuity, appear to be infrequent. The combination of deficits in the electro oculogram and scotopic and flicker electroretinograms suggests that the retinal dystrophy includes defective functioning of retinal pigment epithelial cells and of both rod and cone photoreceptors. The pigmentary retinopathy does not prevent diabetic retinopathy; a single subject had funduscopic and electrophysiologic evidence of both diseases. Current evidence suggests that the mitochondrial DNA A3243G mutation accounts for 0.5% to 2.8% of diabetes. Most ophthalmic and diabetic clinics are therefore likely to contain such patients, who may benefit from identification of the genetic defect causing their disease and from genetic counseling. PMID- 10366078 TI - Low-frequency damped electroretinographic wavelets in young asymptomatic patients with dominant retinitis pigmentosa: a new electroretinographic finding. AB - OBJECTIVE: To describe a previously unreported electroretinographic (ERG) pattern in early retinitis pigmentosa (RP). DESIGN: Two case reports. PARTICIPANTS: Two unrelated young asymptomatic patients with autosomal-dominant retinitis pigmentosa were studied. MAIN OUTCOME MEASURES: Clinical findings and ERG responses were assessed. RESULTS: No ERG responses were detected scotopically with low-luminance stimuli. With increasingly brighter stimuli, a series of three to five low-frequency damped wavelets developed under both scotopic and photopic conditions. The period of the wavelets was 25 to 37 msec. CONCLUSIONS: Low frequency damped ERG wavelets occur in some young asymptomatic patients with autosomal-dominant RP. The ERG pattern suggests that these wavelets are predominantly cone-generated. PMID- 10366079 TI - Oral fluorescein angiography with the confocal scanning laser ophthalmoscope. AB - OBJECTIVE: To evaluate the efficacy of oral fluorescein angiography with a confocal scanning laser ophthalmoscope (SLO) system. DESIGN: Comparative case series. PARTICIPANTS: The authors used a confocal SLO (Heidelberg Retina Angiograph [HRA]) to perform oral fluorescein angiography in 47 patients, 13 of whom were without any retinal disease and 34 with a variety of retinal diseases including macular holes and pucker, inflammatory diseases, retinal vascular diseases, and age-related macular degeneration. The images were also compared to images taken with a fundus camera after intravenous fluorescein injections in patients on whom both studies were done. INTERVENTION: Color fundus photographs were taken of each eye (30 degrees fundus camera) before drinking 4 ml of 25% sodium fluorescein mixed with 60 ml of orange juice. After oral fluorescein ingestion, images of each eye were taken with a fundus camera (TriX film) and the HRA (using 512- x 512-pixel resolution). The images were repeated at 0-, 2.5-, 5 , 7.5-, 10-, 12.5-, 15-, 20-, 25-, and 30-minute intervals. Twenty of the 47 patients underwent intravenous fluorescein angiography performed with the fundus camera. MAIN OUTCOME MEASURE: Images were analyzed by a masked reader, and foveal avascular zone visualization, branch retinal vessel identification, and image quality were scored. Statistical analysis was performed with a t test for paired data with a two-tailed test of significance (alpha = 0.05). RESULTS: Foveal avascular zone was 100% as seen in 16 eyes (47%) in the HRA machine versus 1 eye (2%) in the conventional fundus camera (P < 0.0001). The third-order branch retinal vessels were identified in 59% of eyes in the HRA versus 26% in the fundus camera group (P < 0.0001), and the image quality was considered comparable to an intravenous angiogram in 47% with the HRA versus 9% with the conventional fundus camera (P < 0.0001). CONCLUSIONS: Oral fluorescein angiography using the HRA produces sufficiently detailed images to diagnose, treat, and follow many types of retinal pathology. PMID- 10366080 TI - Computer-assisted, interactive fundus image processing for macular drusen quantitation. AB - PURPOSE: To design and validate a software package to quantitate the area subtended by drusen in color fundus photographs for the conduct of efficient, accurate clinical trials in age-related macular degeneration. DESIGN: Algorithm and software development. Comparisons with manual methodologies. PARTICIPANTS: Evaluation and testing on color fundus photographs from patient records and from eyes enrolled in the Choroidal Neovascularization Prevention Trial. METHODS: Fundus photographs of eyes with drusen were digitized. The green channel was selected for maximum contrast and preprocessed with filtering and shade correction to minimize noise, improve contrast, and correct for illumination and background inhomogeneities. Local thresholding and region-growing algorithms identified drusen. Multiple levels of supervision were incorporated to maximize robustness, accuracy, and validity. Validation studies compared computer-assisted with manual grading by an experienced grader. Intraclass correlation coefficients were calculated as a measure of the concordance between manual and computer assisted fundus gradings. MAIN OUTCOME MEASURES: Drusen area and concordance with manual grading. RESULTS: Automated supervised image analysis offers extreme robustness and accuracy. Most images were segmented with little or no supervision, with processing times on the order of 5 seconds. More complicated images required supervision and a total analysis time varying from 20 seconds to 5 minutes, with most of this time devoted to inspection and comparison. Interactive image processing affords arbitrarily close concordance with manual drusen identification, with calculated intraclass correlation coefficients of 0.92 and 0.93 for comparison of manual with automated, supervised grading by two observers. CONCLUSIONS: Automated supervised fundus image analysis is an efficient, robust, valid technique for drusen quantitation from color fundus photographs. This approach should prove useful in the conduct of efficient accurate clinical trials for age-related macular degeneration. PMID- 10366081 TI - Cytidine-5'-diphosphocholine (citicoline) improves retinal and cortical responses in patients with glaucoma. AB - PURPOSE: To evaluate the effects of cytidine-5'-diphosphocholine (citicoline) on retinal function and on cortical responses in patients with glaucoma. DESIGN: Randomized clinical trial. PARTICIPANTS: Forty patients with open-angle glaucoma were randomly divided into two age-matched groups: citicoline group ([GC] n = 25) and placebo group ([GP] n = 15). METHODS: The GC patients were treated with Neuroton (citicoline, 1000 mg/day intramuscularly) for 60 days; GP patients were treated with placebo (physiologic solution with additives) for 60 days. After 120 days of washout (day 180), the GC patients were divided into two age-matched groups: in 10 patients (GC1 group) the washout was prolonged for a further 120 days; in 15 patients (GC2 group) a second 60-day period of citicoline treatment was followed by a second 120-day period of washout. At day 180, the washout was extended for another 180 days in GP patients. In all subjects, retinal and cortical responses were evaluated by simultaneous recordings of visual evoked potentials (VEPs) and pattern-electroretinograms (PERGs) at baseline, after 60 days, and after 180 days. At day 300, VEPs and PERGs were also evaluated in GC1 patients, and at 240 and 360 days in GC2 and GP patients. MAIN OUTCOME MEASURES: Visual evoked potential parameters (P100 latency and N75-P100 amplitude); PERG parameters (P50 latency and P50-N95 amplitude); and intraocular pressure. RESULTS: The GP patients displayed similar VEP and PERG parameters in all examinations performed. In GC patients, the treatment with citicoline induced a significant (P < 0.01) improvement of VEP and PERG parameters, and their values were significantly different (P < 0.01) with respect to those of GP patients (P < 0.01). Visual evoked potentials and PERGs, recorded in GC patients after washout, revealed that although there was a worsening trend, the electrophysiologic improvement was still maintained. After a second period of washout, GC1 patients had VEP and PERG parameters similar (P > 0.05) to baseline ones and to those of GP patients. In GC2 patients, a second period of citicoline treatment induced a further (P < 0.01) improvement of VEP and PERG parameters CONCLUSION: Citicoline may induce an improvement of the retinal and of the visual pathway function in patients with glaucoma. PMID- 10366082 TI - Efficacy of apraclonidine 1% versus pilocarpine 4% for prophylaxis of intraocular pressure spike after argon laser trabeculoplasty. AB - OBJECTIVE: The authors compared the efficacy of apraclonidine 1% versus pilocarpine 4% prophylaxis of post-argon laser trabeculoplasty (ALT) intraocular pressure (IOP) spike. DESIGN: Prospective randomized clinical trial. PARTICIPANTS: Two hundred twenty-eight eyes of 228 patients with primary open angle glaucoma undergoing ALT were studied. INTERVENTION: Patients were given 1 drop of either apraclonidine 1% (n = 114) or pilocarpine 4% (n = 114) 15 minutes before ALT. MAIN OUTCOME MEASURES: Peri-ALT IOPs and incidences of post-ALT IOP spikes at 5 minutes, 1 hour, and 24 hours were compared between the two groups. RESULTS: The two groups were similar in age, race, and medical dependency. Post ALT mean IOPs at 5 minutes, 1 hour, and 24 hours were significantly lower than pre-ALT mean IOPs in both apraclonidine (P < 0.001) and pilocarpine (P < 0.001) groups. Incidences of IOP spikes greater than 1, 3, and 5 mmHg at 1 hour post-ALT were 21.1%, 14.9%, and 8.8% for the apraclonidine group and 12.3%, 5.3%, and 4.4% for the pilocarpine group (P = 0.076, 0.015, and 0.18 chi-square test). In the apraclonidine prophylaxis group, patients on long-term apraclonidine showed significantly higher incidence of post-ALT IOP spike than the patients without such long-term apraclonidine use (35.7%, 15 of 42 eyes, vs. 12.5%, 9 of 72 eyes; P = 0.003). In addition, peri-ALT pilocarpine prophylaxis tended to be less effective in patients undergoing long-term pilocarpine therapy but without statistical significance (17.4%, 8 of 46 eyes, vs. 9.4%, 6 of 64 eyes; P = 0.17). CONCLUSION: Peri-ALT pilocarpine 4% was at least as effective as, if not more effective than, apraclonidine 1% in post-ALT IOP spike prophylaxis. Peri-ALT apraclonidine prophylaxis was not effective in patients on long-term apraclonidine, and peri-ALT pilocarpine prophylaxis tended to be less effective in patients undergoing long-term pilocarpine therapy. Pilocarpine 4% can be considered as a first-choice drug for post-ALT IOP spike prophylaxis, especially in patients under treatment with apraclonidine. PMID- 10366083 TI - The comparative benefits of glaucoma filtering surgery with an electric-pulse targeted drug delivery system demonstrated in an animal model. AB - PURPOSE: To evaluate the safety and effectiveness of glaucoma filtering surgery performed with the adjunctive use of bleomycin administered in conjunction with electric pulses (EP). DESIGN: Experimental study in rabbits. CONTROLS AND METHODS: Trabeculectomies were performed on pigmented rabbits (2 to 2.5 kg) using the following adjunctive treatments: 5 microM of topical bleomycin and EP (5V, 50 msec, 8 pulses) (group A: B+E+, n=15); bleomycin but no EP (group B: B+E-, n=15); 5 ,uM mitomycin C (MMC) and EP (group C: M+E+, n= 10); MMC but no EP (group D: M+E-, n=10); EP alone (group E: E+, n=10); and no adjunctive treatment (group F: E-, negative control, n=10). MAIN OUTCOME MEASURES: Intraocular pressure (IOP) was measured regularly for 60 days after the operation. Bleb formation and the condition of the conjunctiva, cornea, and retina were also regularly evaluated. Histologic studies were performed by light microscopy, and retinal functions were evaluated by electroretinography. RESULTS: Postoperative IOP was significantly lower than the preoperative level in all the animals until day 7. However, in groups E and F (the negative control) it returned to the preoperative level after day 7, and in groups B, C, and D after 15 days. The IOP of group A remained lower even on day 40. The average amount IOP was lowered or increased on day 20 was 6.4 mmHg (P < 0.05) in group A; -0.2 mmHg in group B; +1.2 mmHg in group C; and 3.25 mmHg in group D. The survival rate of the filtering blebs on day 20 was significantly higher in group A than in the other groups. Clinical and histologic studies uncovered no pathologic findings in any intra- or paraocular tissues. Electroretinographic evaluation of retinal function in group A showed no apparent change over the 60 days of the study. CONCLUSION: Glaucoma filtering surgery in rabbits with the adjunctive use of bleomycin in conjunction with EP significantly lowered IOP for up to 40 days without clinically, morphologically, or functionally harming intraocular tissues. PMID- 10366084 TI - Longitudinal changes in optic disc topography of adult patients after trabeculectomy. AB - OBJECTIVE: To study longitudinal changes in optic disc topography after trabeculectomy in adult patients. DESIGN: Prospective case series. PARTICIPANTS: Twenty-five eyes of 25 patients undergoing trabeculectomy were enrolled. INTERVENTION: Images of the optic disc were obtained preoperatively and approximately 2 weeks, 4 months, and 8 months after surgery by use of a confocal scanning laser ophthalmoscope (Heidelberg Retina Tomograph). MAIN OUTCOME MEASURES: The topographic optic disc parameters (cup volume, cup area, rim volume, rim area, cup-disc area ratio, mean cup depth, maximum depth, cup shape, and height variation contour) were measured automatically for each image with the Heidelberg Retina Tomograph Software (version 1.11). RESULTS: Approximately 2 weeks after surgery, the mean preoperative intraocular pressure (IOP) of 19.3 mmHg (SD, 6.4 mmHg) decreased to 6.0 mmHg (SD, 3.6 mmHg), cup volume and mean cup depth decreased, height variation contour increased, and the cup shape parameter became more negative. Approximately 4 months after surgery, mean IOP was 9.7 mmHg (SD, 4.2 mmHg), and the only statistically significant change from preoperative values of optic disc parameters was in the cup shape measure. Approximately 8 months after surgery, there was no statistically significant change in any of the optic disc parameters compared with preoperative values, although IOP was 10.4 mmHg (SD, 5.9 mmHg). CONCLUSIONS: Changes in the optic disc that may be present 2 weeks after a trabeculectomy do not appear to persist 4 and 8 months later in eyes with advanced glaucomatous optic nerve damage, except for cup shape, which was different from preoperative values at 4 months but not at 8 months. PMID- 10366085 TI - Phacotrabeculectomy: limbus-based versus fornix-based conjunctival flaps in fellow eyes. AB - OBJECTIVE: To compare the effectiveness of limbus-based and fornix-based conjunctival flaps in fellow eyes of the same patients undergoing combined trabeculectomy with phacoemulsification. DESIGN: Prospective, nonrandomized comparative (fellow eye) study. PARTICIPANTS: Forty-four patients and 88 fellow eyes. INTERVENTION: Limbus-based conjunctival flap with phacotrabeculectomy was performed in one eye, and a fornix-based conjunctival flap with phacotrabeculectomy was performed in the fellow eyes of the same patients. The patients were followed up for a minimum of 1 year postoperatively for each eye. MAIN OUTCOME MEASURES: Preoperative and postoperative visual acuity, intraocular pressure, number of antiglaucoma medications, interventions, and complications were studied. RESULTS: At last follow-up visit, visual acuity improved to 20/40 or better in 88.6% of the limbus-based group and 79.6% of the fornix-based group. Preoperatively, the mean intraocular pressure in the limbus-based group was 21.4 +/- 4.8 mmHg on a mean of 2.4 +/- 1.2 glaucoma medications; in the fornix-based group, it was 21.4 +/- 4.3 mmHg on a mean of 2.3 +/- 1.1 medications. Mean intraocular pressure decreased to 15.3 +/- 3.3 mmHg (P < 0.01) on a mean of 0.2 +/- 0.5 glaucoma medications in the limbus-based group (P < 0.01). In the fornix based group, mean intraocular pressure at last follow-up visit decreased to 15.3 +/- 4.7 mmHg (P < 0.01) on a mean of 0.2 +/- 0.5 medications (P < 0.01). Postoperative interventions and complications were not statistically different between the two groups. CONCLUSION: With phacotrabeculectomy, limbus-based and fornix-based conjunctival flaps are equally effective in improving visual acuity and lowering intraocular pressure. This variation in conjunctival flap orientation was equally effective in fellow eyes of the same patients, with no difference in postoperative complications or outcomes. PMID- 10366086 TI - Nonmechanical corneal trephination with the excimer laser improves outcome after penetrating keratoplasty. AB - OBJECTIVE: To assess the impact of nonmechanical trephination on the outcome after penetrating keratoplasty (PK). DESIGN: Prospective, randomized, cross sectional, clinical, single-center study. PATIENTS: A total of 179 eyes of 76 females and 103 males, mean age at the time of surgery 50.6 +/- 18.5 (range, 15 83) years. Inclusion criteria were (1) time interval from October 1992 to December 1997; (2) one surgeon (GOHN); (3) primary central PK; (4) Fuchs dystrophy (diameter, 7.5 mm) or keratoconus (diameter, 8.0 mm); (5) graft oversize, 0.1 mm; (6) no previous intraocular surgery; and (7) 16-bite double running diagonal suture. INTERVENTION: In a randomized fashion, eyes were assigned either to trephination with the 193-nm Meditec excimer laser (manually guided beam in patients, automated rotation device of artificial anterior chamber in donors) along metal masks with eight orientation teeth/notches (EXCIMER: 53 keratoconus, 35 Fuchs dystrophy; mean follow-up, 37 +/- 16 months) or with a hand held motor trephine (Microkeratron; Geuder) ( CONTROL: 53 keratoconus, 38 Fuchs dystrophy; mean follow-up, 38 +/- 14 months). Subjective refractometry (trial glasses), standard keratometry (Zeiss), and corneal topography analysis (TMS-1; Tomey) were performed before surgery, before removal of the first suture (15.2 +/ 4.2 months), and after removal of the second suture (21.4 +/- 5.6 months). MAIN OUTCOME MEASURES: Keratometric and topographic net astigmatism as well as refractive cylinder; keratometric and topographic central power; best-corrected visual acuity (VA); surface regularity index (SRI), surface asymmetry index (SAI), and potential visual acuity (PVA) of the TMS-1. RESULTS: Before suture removal, mean refractive/keratometric/topographic astigmatism did not differ significantly between EXCIMER (2.5 +/- 1.8 diopters [D]/3.4 +/- 2.8 D/4.7 +/- 3.1 D) and CONTROL groups (3.0 +/- 1.8 D/3.7 +/- 2.4 D/4.3 +/- 2.1 D). After suture removal, respective values were significantly lower in the EXCIMER group (2.8 +/- 2.0 D/3.0 +/- 2.1 D/3.8 +/- 2.6 D) than in the CONTROL group (4.2 +/- 2.4 D/6.1 +/- 2.7 D/6.7 +/- 3.1 D) (P < 0.0009). In the EXCIMER versus CONTROL group, mean VA increased from 20/100 versus 20/111 (P > 0.05) before surgery, to 20/31 versus 20/38 before (P = 0.001) and to 20/28 versus 20/39 (P < 0.00001) after suture removal. Mean spherical equivalent was significantly less myopic in the EXCIMER group before (-0.9 +/- 3.6 D vs. -2.6 +/- 3.4 D) (P = 0.01) and after suture removal (-1.4 +/- 3.1 D vs. -2.4 +/- 3.5 D) (P = 0.02). Mean SRI (P = 0.04) and PVA (P = 0.007) were significantly more favorable in the EXCIMER versus CONTROL group after suture removal (0.91 +/- 0.45 and 0.82 +/- 0.15 vs. 1.05 +/- 0.46 and 0.73 +/- 0.18). CONCLUSIONS: Postkeratoplasty results seem to be superior using nonmechanical excimer laser trephination. Thus, this methodology is recommended as the procedure of first choice in avascular corneal pathologies requiring PK. PMID- 10366087 TI - Risk factors in microbial keratitis leading to penetrating keratoplasty. AB - OBJECTIVE: To determine the characteristics of infectious corneal ulcers at the time of presentation to the cornea specialist associated with a favorable response to medical therapy versus a poor outcome manifested by the need for penetrating keratoplasty for therapy or visual rehabilitation. DESIGN: Retrospective, case-control study. PARTICIPANTS: A total of 162 patient records were reviewed, including the study group of 30 patients and the control group of 132 patients. INTERVENTION: A retrospective review of all cases of microbial keratitis presenting to the Cornea Service between January 1, 1989 and December 31, 1995 was conducted. The cases were divided into two groups. The study group consisted of patients with microbial keratitis who failed medical therapy and required penetrating keratoplasty. The control group included patients with infectious ulcers who responded to medical therapy alone. MAIN OUTCOME MEASURES: The influence of demographics, medical and ocular history, delay in presentation to the primary ophthalmologist or the corneal specialist, topical medications, and contact lens usage were compared. Visual acuity and ulcer characteristics were recorded. The statistical significance was evaluated by the chi-square test for independence and multiple logistic regression. RESULTS: Older age (P=0.001), delay in referral to the corneal specialist (P<0.03), and treatment with topical steroids prior to presentation (P<0.0001) were statistically significant factors associated with the need for penetrating keratoplasty. Steroid use and the delay in referral were correlated. A past history of ocular surgery (P=0.01), poor visual acuity at presentation (P<0.001), and ulcer characteristics, including central location (P<0.0001), large size (P<0.0001), presence of perforation or descemetocele (P<0.0001), limbal involvement (P<0.0001), and hypopyon (P=0.05), were all associated with the need for penetrating keratoplasty. CONCLUSIONS: Older age, delay in referral to the corneal specialist, topical steroid treatment, past ocular surgery, poor vision at presentation, large size, and central location of the ulcer are risk factors for poor outcome of microbial keratitis, as indicated by the need for penetrating keratoplasty. PMID- 10366089 TI - Relationship between apparent accomodation and corneal multifocality in pseudophakic eyes. AB - OBJECTIVE: To investigate whether any association exists between apparent accommodation in pseudophakic eyes and multifocal corneal effects. DESIGN: A prospective observational case series. PARTICIPANTS: A total of 121 eyes of 98 patients who had undergone phacoemulsification and posterior chamber intraocular lens implantation were studied. METHODS: The amount of apparent accommodation was measured using an accommodometer. The degree of corneal multifocality was determined on the corneal topography by measuring the maximum and minimum corneal refractive powers within the pupillary area. Refractive astigmatism, keratometric astigmatism, pupillary diameter, and age were also analyzed. MAIN OUTCOME MEASURES: Apparent accommodation, corneal multifocality, refractive and keratometric astigmatism, pupillary diameter, and age. RESULTS: Multiple regression analysis revealed that corneal multifocality and pupillary diameter had significant positive correlations with apparent accommodation, whereas other explanatory variables showed no relationship with apparent accommodation. CONCLUSION: Multifocal corneal effects contribute to apparent accommodation in pseudophakic eyes. PMID- 10366088 TI - Phase II results of an intraocular steroid delivery system for cataract surgery. AB - OBJECTIVE: To evaluate the safety and efficacy of an intraocular biodegradable polymer dexamethasone drug delivery system (DEX DDS) in treating postoperative inflammation after cataract surgery. STUDY DESIGN: Multicenter, randomized, double-masked, parallel group study comparing two dose levels of the DEX DDS to concurrent placebo and no treatment control subjects. PARTICIPANTS: Ninety patients scheduled to undergo extracapsular cataract extraction with phacoemulsification and intraocular lens implantation participated in the study. INTERVENTION: One or two DEX DDSs, each containing 60 microg of dexamethasone, were placed in the posterior chamber after cataract surgery. Patients receiving the placebo received a DDS composed of the same matrix with no active drug. In vivo rabbit studies have determined that the DEX DDS releases dexamethasone into the anterior chamber (AC) for approximately 7 to 10 days. MAIN OUTCOME MEASURES: The AC cells and the AC flare were assessed over a 60-day postoperative period using slit-lamp examination by masked observers. The number and percent of patients in each treatment group requiring additional postoperative topical anti inflammatory medication were compared. RESULTS: Ninety patients were randomized into 4 treatment groups (30 to the 2 DEX DDS group, 30 to the 1 DEX DDS group, 15 to the placebo DDS group, and 15 to the no treatment group). The control patients required the addition of topical steroids as rescue medication more frequently and sooner than patients receiving DEX DDS (80% vs. 7% at week 2) (P < 0.001). Patients receiving DEX DDS showed a significant reduction in postoperative inflammation as assessed by the combined AC cell and flare scores when compared to the control group from day 3 (P = 0.002) through week 3. The DEX DDS was well tolerated. No clinically significant difference in any safety evaluations, including intraocular pressure, was seen between the DEX DDS-treated and control groups. CONCLUSION: The DEX DDS was safe and effective in suppressing postoperative inflammation after uncomplicated cataract surgery. Additional topical anti-inflammatory drops were not needed for most patients. PMID- 10366090 TI - Eyelid, conjunctival, and corneal findings in sleep apnea syndrome. AB - OBJECTIVE: To determine the prevalence of eyelid, conjunctival, and corneal findings in patients with sleep apnea syndrome (SAS). DESIGN: Case series. PARTICIPANTS: Seventy-two white patients referred for evaluation of suspected SAS. INTERVENTION: Complete examination of eyelids, conjunctiva, and cornea, including videokeratography. MAIN OUTCOME MEASURES: Spearman rank correlations were determined between the respiratory disturbance index (RDI) during night sleep, a value used to diagnose and grade SAS, and tear film break-up time, eyelid distraction distance, presence or absence of ocular irritation symptoms, blepharoptosis, floppy eyelids, lacrimal gland prolapse, keratoconus, and endothelial dystrophy. Each correlation was controlled for age and body mass index. RESULTS: According to the RDI, 44 (61 %) of the 72 patients had SAS. The RDI correlated positively with the eyelid distraction distance (P = 0.05), presence or absence of floppy eyelids (P = 0.01), and lacrimal gland prolapse (P = 0.01), and correlated negatively with tear film break-up time (P = 0.02). None of our patients with floppy eyelids had corneal abnormalities. One patient with SAS had bilateral keratoconus; another had bilateral Fuch endothelial dystrophy. CONCLUSIONS: Sleep apnea syndrome was significantly associated with reduced tear film break-up time, floppy eyelids, and lacrimal gland prolapse. However, ocular irritation symptoms and corneal involvement were rare among patients with SAS. These findings do not confirm previous studies that reported a high prevalence of corneal involvement in floppy eyelid syndrome. PMID- 10366091 TI - Pseudotumor cerebri in children receiving recombinant human growth hormone. AB - PURPOSE: This article represents the first report in the ophthalmology literature of an association between pseudotumor cerebri (PTC) and recombinant human growth hormone (rhGH). DESIGN: Noncomparative case series. PARTICIPANTS: Three children receiving rhGH for short stature with Turner syndrome, Jeune syndrome, or Down syndrome. METHODS: Children underwent full ocular examination. After papilledema was identified, patients underwent lumbar puncture and imaging with either magnetic resonance imaging or computerized tomography. Treatment was under the guidance of the primary physician or neurosurgeon. The rhGH was discontinued in all children. MAIN OUTCOME MEASURES: Visual acuity and evaluation of the optic nerve for resolution of papilledema were followed at each examination. RESULTS: In all three cases, papilledema resolved with the cessation of rhGH, and treatment with acetazolamide or prednisone. Visual acuity was unchanged in case 1, decreased by two to three lines in case 2, and was inconsistent in case 3. One child (case 2) required a ventriculoperitoneal shunt for persistent elevation of intracranial pressure. CONCLUSION: There appears to be a causal relationship between the initiation of rhGH with the development of PTC. Children should have a complete ophthalmic evaluation if they report headache or visual disturbances. Baseline examination with routine follow-up should be instituted when children cannot adequately communicate. PMID- 10366092 TI - Management of moderate-to-severe Marcus-Gunn jaw-winking ptosis. AB - OBJECTIVE: To report the results of levator excision and frontalis suspension for moderate-to-severe Marcus-Gunn jaw-winking ptosis. DESIGN: A retrospective noncomparative case series. PARTICIPANTS: Twenty-four patients with moderate-to severe Marcus-Gunn jaw-winking ptosis (21 unilateral and 3 bilateral) were treated surgically between 1978 and 1997 by one surgeon. INTERVENTION: Levator excision either in the involved eyelid or in both eyelids, followed by bilateral frontalis suspension, was performed. MAIN OUTCOME MEASURES: Postoperative improvement of jaw-winking was determined. The surgical results of ptosis surgery were assessed as good, fair, or poor based on habitual upper eyelid heights and symmetry. RESULTS: Postoperative follow-up periods ranged from 6 months to 153 months, with an average of 36.9 months. After levator excision in a total of 27 eyelids exhibiting jaw-winking, 10 eyelids (37.0%) showed complete resolution of jaw-winking, and 13 eyelids (48.2%) showed mild winking (1 mm or less) on the lateral jaw movement only (functionally and cosmetically not a problem). In four eyelids (14.8%), these results were not recorded. In the group of five patients undergoing bilateral frontalis suspension and levator excision only on the involved side, final results were good in two patients (40%) and poor in three (60%). Of the 19 patients who underwent bilateral levator excision, final results were good in 13 (68.4%) and fair in 6 (31.6%). CONCLUSIONS: For moderate-to severe jaw-winking ptosis, bilateral frontalis suspension after bilateral levator excision generally provided satisfactory correction of both jaw-winking and ptosis. PMID- 10366094 TI - A new device for ocular surgical training on enucleated eyes. AB - OBJECTIVE: To develop a reliable inexpensive device for teaching ocular surgical procedures and practicing experimental techniques on enucleated eyes. DESIGN: Teaching device trial. PARTICIPANTS: Thirty enucleated porcine eyes. METHODS: A Plexiglas ocular bulb holder was secured with its base support to a polyvinylchloride pillar on a modified polystyrene trial head. MAIN OUTCOME MEASURE: The convenience and reproducibility of both laser and surgical ocular techniques performed with this new device were evaluated. RESULTS: This model allows curvilinear capsulorrhexis and phacoemulsification of porcine lenses through a corneal tunnel incision and insertion of a soft foldable acrylic intraocular lens into the capsular bag. Argon and neodymium:YAG laser iridotomy and retinal argon laser photocoagulation can also be performed with this model. CONCLUSIONS: This inexpensive device is useful for teaching both surgical and laser ocular procedures. PMID- 10366093 TI - Distensible venous malformations of the orbit: clinical and hemodynamic features and a new technique of management. AB - OBJECTIVE: To investigate distensible venous malformations of the orbit (DVMO) as part of a spectrum of orbital vascular malformations, including some that involved periorbital skin, extraorbital sites (central nervous system or nasal sinuses), or combinations of these. The authors also investigated the effectiveness of a new technique of management for selected cases. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Thirty patients had distensible venous anomalies, of which four were combined distensible venous lymphatic vascular malformations. Distensible lesions were defined as those showing clinical or radiographic expansion with Valsalva maneuver or when the head was placed in a dependent position. These lesions were then classified as superficial (anterior to the equator of the globe), deep (posterior to the globe's equator), combined (deep and superficial), or complex (with intracranial or major extraorbital involvement). INTERVENTION: Surgery was performed on 15 patients (50%), mainly for pain or for cosmetic indications. Six patients underwent this new technique, which involved intraoperative direct venography with control of outflow via pressure at the superior or inferior orbital fissure. The venous malformation was then embolized (by use of cyanoacrylate glue) and excised. RESULTS: The mean age at presentation was 28.2 years (range, 8 months to 75 years). Sixty-six percent of cases involved the left orbit. Superior and medial orbital involvement was most common. Three cases (10%) were classified as superficial, and 13 (43%) as deep. Six patients (20%) had combined superficial and deep components. Eight (27%) had major extraorbital involvement (4 intracranial, 2 facial, and 2 paranasal sinus). Direct venography demonstrated complex multichannel anomalies draining to various sites, including the face and pterygopalatine fossa, without necessarily having a direct connection to the major orbital venous circulation. CONCLUSIONS: Distensible venous malformations of the orbit are part of a spectrum of developmental venous malformations that may be localized to the orbit or involve it as part of a more extensive lesion. The authors describe their clinical and radiologic features and report a new technique of management for selected cases. This method of vascular isolation and embolization of lesions may greatly facilitate excision. PMID- 10366095 TI - Benefits of stereolithography in orbital reconstruction. AB - OBJECTIVE: To describe the benefits of the stereolithography (SLA) modeling system in the evaluation and surgical planning of selected bony orbital pathology. DESIGN: Two case reports. PARTICIPANTS: One patient presented with a displaced left orbital roof fracture into his orbit causing globe compression and binocular vertical diplopia. A second patient underwent removal of his right orbital floor, medial wall, and inferior portion of his lateral wall during excision of a cylindrical cell papilloma of the paranasal sinuses. Postoperatively, he suffered from globe ptosis and binocular oblique diplopia. INTERVENTION: Stereolithographic models of the patients' orbits were obtained from computed tomography data to better assess the bony orbital pathology. In the second patient, the model was used as a template to create a temporary custom fit prosthesis to repair the defect of his orbital walls. RESULTS: The SLA models were useful in evaluating the dimensions of the bony defects and in preoperative surgical planning. Intraoperatively, the SLA models facilitated orbital surgical rehabilitation. Postoperatively, both patients noted resolution of their diplopia after reconstruction of more normal bony anatomy. CONCLUSIONS: In selected cases, SLA offers highly accurate models of the bony orbit for preoperative evaluation, surgical planning, and teaching and can act as a template for custom prosthesis manufacturing. This technology increases the orbital surgeon's options in managing complex orbital pathology. PMID- 10366096 TI - Prepartum mixed type cavernous-capillary hemangioma arising in nevus flammeus. AB - OBJECTIVE: Capillary hemangioma may appear de novo and involute during the first decade of life, but rarely during pregnancy. This study describes the clinical and histologic findings of an eyelid mixed type cavernous-capillary hemangioma arising in a nevus flammeus and discusses the differential diagnosis of this lesion. STUDY DESIGN: Clinicopathologic case report. INTERVENTION: A reddish, protruding eyelid mass arising from a nevus flammeus at the eyelid margin in a 26 year-old woman was monitored during her pregnancy. Postpartum, the mass was excised and examined histologically. RESULTS: The lobulated tumor recurred during the second pregnancy and partially regressed following delivery. It was composed of mixed elements of cavernous and capillary hemangioma that superficially resembled Kaposi sarcoma, set against the background of a nevus flammeus. CONCLUSION: The differential diagnosis of discrete prepartum vascular tumor arising in nevus flammeus includes mixed capillary-cavernous hemangioma, pseudo Kaposi sarcoma, granuloma gravidarum, and angiodermatitis. A common stimulus during pregnancy may be the inciting factor for the development of these tumors. PMID- 10366097 TI - Giant cell angiofibroma of the orbit and eyelid. AB - PURPOSE: To report the clinicopathologic features of a newly recognized tumor, giant cell angiofibroma. DESIGN: Observational case series. MAIN OUTCOME MEASURES: Clinical and histopathologic features of giant cell angiofibroma. METHODS: Light and electron microscopy and immunohistochemistry of five cases of giant cell angiofibroma. RESULTS: A total of five patients (4 women and 1 man) are described: two presented with a painless mass in the eyelid, two with a mass in the orbit, and one presented with a conjunctival lesion. All lesions were well demarcated with no capsule and were composed of blood vessels, a patternless spindle-shaped cell proliferation with a solid and pseudovascular appearance, and multinucleated giant cells. Both spindle-shaped and giant tumor cells were intensely positive for CD34 and vimentin. CONCLUSION: Giant cell angiofibroma resembles solitary fibrous tumor and giant cell fibroblastoma and should be considered in the differential diagnosis of spindle-cell tumors in the eyelid, orbit, and conjunctiva. PMID- 10366098 TI - The genetics revolution and the assault on rheumatoid arthritis. PMID- 10366099 TI - Psoriatic arthritis: a unified concept twenty years on. PMID- 10366100 TI - Dominant-negative p53 mutations in rheumatoid arthritis. AB - OBJECTIVE: Studies were performed to determine if p53 mutations identified in rheumatoid arthritis (RA) synovial tissue are dominant negative. METHODS: Site directed mutagenesis was used to produce 2 RA-derived mutants: asparagine- >serine at codon 239 (N239S) and arginine-->stop at codon 213 R213*). HS68 dermal fibroblasts were transfected with either empty vector, wild-type p53 cDNA (wt), or the N239S or R213* mutant p53 cDNA clones. Interleukin-6 (IL-6) and bax gene expression were determined by Northern blot analysis. Bax transcription was determined using a bax promoter/reporter gene construct (bax-luc). RESULTS: Transfection of HS68 cells with wt increased bax mRNA levels. This process was blocked by cotransfection with either mutant. The mutant p53 genes also increased IL-6 gene expression. Low levels of bax promoter activity were detected in HS68 cells co-transfected with bax-luc and empty vector, N239S, or R213*, indicating that the RA mutants lacked transcriptional activity. Transfection with wt and bax luc led to a 10-fold increase in luciferase expression. When the wt gene was cotransfected with either of the mutants, there was a dose-dependent inhibition of bax promoter activity. CONCLUSION: These data indicate that at least 2 of the p53 mutants identified in RA joint samples are dominant negative and suppress endogenous wild-type p53 function. PMID- 10366101 TI - Interleukin-1beta, interleukin-1 receptor antagonist, interleukin-4, and interleukin-10 gene polymorphisms: relationship to occurrence and severity of rheumatoid arthritis. AB - OBJECTIVE: To test if interleukin-1beta (IL-1beta), IL-1 receptor antagonist (IL 1Ra), IL-4, or IL-10 gene polymorphisms could be used as markers of susceptibility or severity in rheumatoid arthritis (RA). METHODS: The study included 108 patients with early RA followed up for 2 years and 128 healthy controls. From genomic DNA, 6 polymorphisms in genes for IL-1beta, IL-1Ra, IL-10, and IL-4 were typed. Allelic frequencies and carriage rates were compared between RA patients and controls, between patients with erosive and nonerosive RA, and between patients with or without sustained remission. RESULTS: The RP1 allele of the IL-4 gene was found with a significantly higher frequency in RA patients compared with controls. The combination of an RA-related HLA-DR allele expressing shared epitope and the presence of allele E2 in IL-1beta exon 5 was found to expose patients to an increased risk of erosive disease, with an odds ratio of 8.20 (95% confidence interval 2.59-25.84, P < 0.0001). No significant association was observed between polymorphisms and the occurrence of sustained remission. CONCLUSION: This report, for the first time, indicates an association between RA and a polymorphic IL-4 gene sequence located in 5q31-33. In addition, the results show the prognostic value of a polymorphism in IL-1beta exon 5, which allowed prediction of erosive disease with a specificity of 91.8% in 42.1% of patients. Although these observations are very interesting, they have to be considered preliminary and will need to be confirmed. PMID- 10366102 TI - Polymorphic haplotypes of the interleukin-10 5' flanking region determine variable interleukin-10 transcription and are associated with particular phenotypes of juvenile rheumatoid arthritis. AB - OBJECTIVE: To determine the distribution of the interleukin-10 (IL-10) 5' flanking region haplotypes in children with arthritis and in controls, and to investigate the functional significance of each haplotype. METHODS: Sequence specific oligonucleotide probing was used to determine haplotype frequency. Transient transfection studies were used to investigate the transcription of reporter genes driven by each haplotype. Whole blood cultures were performed to assess IL-10 production by each genotype. RESULTS: Patients with arthritis involving >4 joints were more likely to have a genotype with an ATA haplotype than those whose arthritis remained restricted to <4 joints. This ATA haplotype was associated with lower transcriptional activity than the GCC haplotype (P = 0.02), and the ATA/ATA genotype was associated with lower IL-10 production under lipopolysaccharide stimulation than other genotypes (P < 0.02). CONCLUSION: The results of this study demonstrate the functional significance of the ATA haplotype and reveal a significant association of genotypes containing this haplotype with extended oligoarthritis. PMID- 10366103 TI - Association of the course of collagen-induced arthritis with distinct patterns of cytokine and chemokine messenger RNA expression. AB - OBJECTIVE: To quantitate changes in cytokine and chemokine messenger RNA (mRNA) levels during the development and progression of collagen-induced arthritis (CIA) in mice. METHODS: Mice with CIA were scored for arthritis and killed at weekly intervals. Cytokine and chemokine mRNA levels were determined by RNase protection assays of total paw RNA. RESULTS: Arthritic paws exhibited mRNA levels of interleukin-1beta (IL-1beta), IL-2, macrophage inflammatory protein 2 (MIP-2), IL 6, IL-1 receptor antagonist, RANTES, tumor necrosis factor alpha (TNFalpha), TNFbeta, MIP-1alpha, IL-11, transforming growth factor beta1 (TGFbeta1), TGFbeta2, and TGFbeta3 that were increased above mRNA levels in paws of normal, unimmunized mice and that exhibited distinct temporal patterns of mRNA expression. Clinically uninvolved paws also exhibited an increase in mRNA levels of IL-11, RANTES, TNFalpha, TNFbeta, and MIP-1alpha. CONCLUSION: The observed differential temporal cytokine and chemokine mRNA expression patterns suggest that specific cytokines and chemokines have defined roles at various times during the course of autoimmune arthritis. Since most of these cytokines and chemokines are found in human rheumatoid arthritis (RA) synovium and synovial fluids, these findings may have relevance to RA. PMID- 10366104 TI - Hemoglobin protects from streptococcal cell wall-induced arthritis. AB - OBJECTIVE: To investigate the ability of hemoglobin (Hgb), a nitric oxide (NO) scavenger, to deplete excess NO and reduce inflammation and injury in synovial tissue from joints with inflammatory arthritis. METHODS: The severity of streptococcal cell wall-induced arthritis was monitored following administration of Hgb. Plasma nitrite and nitrate levels were measured, and inducible NO synthase (iNOS) and cytokine messenger RNA (mRNA) expression in peripheral blood mononuclear cells (PBMC) and joint tissue were evaluated. RESULTS: Following systemic administration of Hgb to arthritic rats, plasma levels of nitrite and nitrate as well as iNOS mRNA expression in the joints and PBMC were significantly reduced. Moreover, inflammatory cell accumulation and disease pathology in the joint tissue were dramatically attenuated without obvious side effects. Consistent with this reduction in the inflammatory response, cytokine gene expression was decreased in the synovium of Hgb-treated rats. CONCLUSION: Modulation of NO levels through the use of a NO scavenger, Hgb, influences the development and severity of arthritis. These findings suggest that depletion of excess NO by NO scavengers provides a prototype for further exploration of potential treatments for chronic arthritis. PMID- 10366105 TI - Kinetics of aggrecanase- and metalloproteinase-induced neoepitopes in various stages of cartilage destruction in murine arthritis. AB - OBJECTIVE: Two major cleavage sites, one mediated by metalloproteinases (MMPs) and the other by an as-yet unidentified enzyme termed aggrecanase, have been observed in aggrecan. To learn more about the relative contribution of these enzymes during cartilage degradation, this study assessed the occurrence of both specific neoepitopes in cartilage during murine arthritis and examined the correlation between neoepitope formation and different aspects of cartilage damage. METHODS: Reversible cartilage damage was induced in mice in the zymosan induced arthritis (ZIA) model, partly irreversible cartilage damage in the antigen-induced arthritis (AIA) model, and irreversible, destructive cartilage damage in the collagen-induced arthritis (CIA) model. Immunolocalization techniques were used to detect the specific C-terminal neoepitopes VDIPEN (MMPS) and NITEGE (aggrecanase). RESULTS: In normal cartilage from young adult mice, no VDIPEN epitopes were detected, but a limited amount of NITEGE epitopes were already present. During the early phase of proteoglycan (PG) depletion, NITEGE expression was raised substantially in all arthritis models. VDIPEN epitopes were not detected in this early phase of cartilage destruction. When PG depletion progressed toward advanced cartilage damage, VDIPEN epitopes were induced. During ZIA, minimal induction of VDIPEN was observed, whereas in AIA, strong, but partly reversible, VDIPEN staining was evident, and in CIA, an extensive presence and persistence of the MMP-induced neoepitope was seen. When VDIPEN epitopes were intensely present, NITEGE epitopes were greatly reduced at that site in the cartilage. CONCLUSION: Presence of VDIPEN epitopes in cartilage correlated with severe cartilage damage, but these epitopes were not detected during early PG degradation. This suggests a limited role for VDIPEN-inducing MMPs in early PG degradation during murine arthritis. In contrast, aggrecanase epitopes were induced before the appearance of VDIPEN epitopes, but they disappeared with progression of cartilage damage. PMID- 10366106 TI - Specificity of inhibition of matrix metalloproteinase activity by doxycycline: relationship to structure of the enzyme. AB - OBJECTIVE: To investigate the inhibition of matrix metalloproteinase 1 (MMP-1), MMP-8, and MMP-13 by doxycycline, and to determine whether the variable hemopexin like domain of each MMP was responsible for the differences in susceptibility to doxycycline inhibition among these collagenases. METHODS: Recombinant human MMP-1 (collagenase 1), MMP-8 (collagenase 2), and MMP-13 (collagenase 3), truncated forms of MMP-8 and MMP-13 lacking the hemopexin-like domain, and a mutant form of truncated MMP-13 were used in these studies. The activity of the full-length MMP in the presence of doxycycline was tested against type II collagen, a natural substrate for the enzymes. A small peptolide substrate was used to determine which structural features of the MMPs were related to sensitivity to doxycycline inhibition. RESULTS: The activity of MMP-13 and MMP-8 against type II collagen was inhibited by 50-60% by 30 microM doxycycline, while that of MMP-1 was inhibited only 18% by 50 microM doxycycline. In contrast, in experiments with the peptolide substrate, neither full-length nor truncated MMP-13 was inhibited until the concentration of the drug exceeded 90 microM. MMP-8 and truncated MMP-8 were sensitive to inhibition by 30 microM doxycycline, while MMP-1 was slightly inhibited (14%) by 90 microM doxycycline. For MMP-8, inhibition was reversible upon dilution and was independent of the order in which the reagents were added. Kinetic analysis of the inhibition constant (K(i)) of MMP-8 (K(i) = 36 microM) and truncated MMP-8 (K(i) = 77 microM) indicated that inhibition was noncompetitive. CONCLUSION: Significant inhibition of MMP-13 and MMP-8 activity against collagen occurred in vitro at concentrations that were near the concentrations achieved in serum after oral dosing. Studies with truncated enzymes and 2 substrates suggest that doxycycline disrupts the conformation of the hemopexin-like domain of MMP-13 and the catalytic domain of MMP-8. PMID- 10366107 TI - Collagenase 3 production by human osteoarthritic chondrocytes in response to growth factors and cytokines is a function of the physiologic state of the cells. AB - OBJECTIVE: We investigated the response of human osteoarthritic (OA) chondrocytes, in terms of collagenase 3 production, to growth factors and cytokines involved in the anabolism and catabolism of articular cartilage, and explored the major signaling pathways leading to its up-regulation. METHODS: Human OA chondrocytes were treated with the following factors: the proinflammatory cytokine interleukin-1beta (IL-1beta), the growth factors basic fibroblast growth factor (bFGF), platelet-derived growth factor BB (PDGF-BB), parathyroid hormone (PTH), insulin-like growth factor 1 (IGF-1), transforming growth factor gamma1 (TGFbeta1), and TGFbeta2, the protein kinase (PK) activator antagonists for PKC, PKA, and PKG pathways, and phospholipase A2 and tyrosine kinases, as well as the antiinflammatory cytokines IL-4, IL-10, and IL-13. Collagenase 3 expression and synthesis were determined. Comparison was made with collagenase 1. RESULTS: The human OA chondrocyte population could be divided into 2 categories: the L chondrocytes, showing low collagenase 3 basal synthesis levels and high sensitivity to IL-1beta stimulation; and the H chondrocytes, high collagenase 3 basal synthesis levels and low IL-1beta inducibility. In L chondrocytes, all growth factors stimulated collagenase 3 production. In H chondrocytes, PTH, IGF-1, and TGFbeta had little or no impact; bFGF slightly stimulated it and PDGF-BB showed the same pattern as in the L chondrocytes. The effects of all growth factors, except TGFbeta, on collagenase 1 synthesis followed those of collagenase 3, albeit to a higher degree. Interestingly and unlike collagenase 3, the effects of TGFbeta on collagenase 1 could not be related to the state of the cells, but rather, depended on the isoform. Indeed, TGFbeta2 did not induce collagenase 1 synthesis, whereas TGFbeta1 stimulated it. Among the PK activators tested, phorbol myristate acetate was the strongest inducer, suggesting a major involvement of the PKC pathway. IL-13 inhibited collagenase 3 production, IL-4 had little effect, and IL-10 had none. CONCLUSION: This study shows that collagenase 3 production in human OA chondrocytes depends on the physiologic state of the cell. TGFbeta might be responsible for the change in cells from the L to the H state. Importantly, our in vitro data implicate TGFbeta2 as a possible in vivo agent capable of specifically triggering collagenase 3 production over that of collagenase 1 in OA cartilage. PMID- 10366108 TI - Treatment with calcitonin suppresses the responses of bone, cartilage, and synovium in the early stages of canine experimental osteoarthritis and significantly reduces the severity of the cartilage lesions. AB - OBJECTIVE: To relate the rate of bone resorption to serum levels of both hyaluronan (HA) and antigenic keratan sulfate (KS) in canine experimental osteoarthritis (OA) and to evaluate the effects of calcitonin on these parameters and the OA lesions of the unstable knee. METHODS: Twenty-two dogs underwent anterior cruciate ligament transection (ACLT) and 6 dogs underwent sham operation. Urinary pyridinium crosslinks were quantified by high-performance liquid chromatography. Immunoassays quantified hyaluronan (HA) and antigenic KS. Macroscopic and histologic OA lesions were scored. Calcitonin treatment was started on day 14 postsurgery and stopped on either day 49 or day 104 postsurgery. Control dogs and all treated dogs were killed on day 105. RESULTS: All ACLT joints developed OA. In contrast to sham-operated animals, all operated dogs exhibited an early and sustained rise in the levels of their urinary and serum markers. Calcitonin markedly reduced the levels of these markers and the severity of OA lesions. Furthermore, the longer the period of calcitonin therapy, the lower the score of the OA lesions. CONCLUSION: Bone, synovium, and articular cartilage all appear to be involved in the state of hypermetabolism that develops in unstable joints. Furthermore, the rate of bone resorption increases markedly in the early stages of this OA model and is likely to contribute to cartilage breakdown. Since calcitonin reduced the severity of OA changes, this form of therapy may have benefits for humans who have recently experienced a traumatic knee injury. PMID- 10366109 TI - Production of type 2 cytokines by CD8+ lung cells is associated with greater decline in pulmonary function in patients with systemic sclerosis. AB - OBJECTIVE: This study addresses the hypothesis that a profibrotic pattern of cytokines is produced in the lungs of patients with systemic sclerosis (SSc) and causes fibrosis. METHODS: Using a reverse transcriptase-polymerase chain reaction technique, interleukin-4 (IL-4), IL-5, and interferon-gamma (IFNgamma) messenger RNA (mRNA) were measured in unseparated CD8+ and CD4+ bronchoalveolar lavage (BAL) cells from SSc patients and healthy controls. To confirm the results, CD8+ T cells were cloned from BAL fluids, and the pattern of cytokine mRNA made by these cells was determined. Serial pulmonary function tests were done. RESULTS: BAL cells from healthy controls made IFNgamma mRNA, with no or little IL-4 or IL 5 mRNA. In contrast, BAL cells from the majority of SSc patients made IL-4 and/or IL-5 mRNA, with or without approximately equal amounts of IFNgamma mRNA. This pattern of cytokines was made by CD8+ T cells, which were increased in the lungs of these SSc patients. Patients whose BAL cells made this type 2 pattern of cytokine mRNA had a significant decline in forced vital capacity over time after the BAL, whereas patients whose BAL cells made IFNgamma mRNA alone did not. Both wild-type and an alternative splice variant of IL-4 mRNA were increased in BAL cells from SSc patients. Both forms of IL-4 stimulated alpha2(I) collagen mRNA in human dermal and lung fibroblasts. CONCLUSION: The type 2 pattern of cytokine mRNA produced by BAL cells from SSc patients differs from unopposed IFNgamma production found in healthy BAL cells. This production of type 2 cytokine mRNA by CD8+ T cells is associated with a significant decline in lung function over time, which suggests a pathologic role for these T cells in interstitial fibrosis in SSc. PMID- 10366110 TI - An immunodominant epitope on DNA topoisomerase I is conformational in nature: heterogeneity in its recognition by systemic sclerosis sera. AB - OBJECTIVE: To characterize an immunodominant epitope recognized by anti-DNA topoisomerase I (topo I) antibody, a major autoantibody in sera of patients with systemic sclerosis (SSc). METHODS: Topo I fragments were generated as fusion proteins using a bacterial expression system as well as polypeptides translated in vitro using a eukaryotic expression system. Reactivities to the 2 preparations of recombinant topo I polypeptides in anti-topo I-positive sera from SSc patients of varied ethnic backgrounds were examined by immunoblotting, immunoprecipitation, and/or enzyme-linked immunosorbent assay. RESULTS: The fragment encoding amino acids 489-573 of topo I was recognized by 98 of 100 anti topo I-positive SSc sera. Both carboxyl- and amino-terminal deletion studies as well as competitive inhibition assays using topo I synthetic peptides showed that a region of > or =52 amino acids (512-563) was necessary for recognition by anti topo I antibodies. The minimum epitope region and conformation required for this reactivity were variable among sera from Caucasian, African American, Japanese, and Choctaw SSc patients. CONCLUSION: An immunodominant epitope recognized by anti-topo I autoantibody is located in the region of amino acids 489-573 of the topo I protein and is largely conformational in nature. The recognition pattern of this region by anti-topo I-positive sera is heterogeneous and is influenced by ethnic background. PMID- 10366111 TI - Polymorphism of beta2-glycoprotein I at codons 306 and 316 in patients with systemic lupus erythematosus and antiphospholipid syndrome. AB - OBJECTIVE: To determine the frequency of mutations in the phospholipid binding domain of beta2-glycoprotein I (beta2GPI) in patients with systemic lupus erythematosus (SLE) and/or antiphospholipid syndrome (APS), and to analyze the clinical correlations of such mutations with thromboembolic complications. METHODS: Exons 7 and 8 of beta2GPI, which encode for its fifth domain, were amplified by polymerase chain reaction, and the presence of mutations was determined by restriction digestion and single-strand conformation polymorphism analysis. A clinical correlation with these mutations and the presence of antiphospholipid antibodies (aPL), lupus anticoagulant (LAC), anti-beta2GPI antibody, and the development of thromboembolic complications was performed using chi-square and Fisher's exact tests. RESULTS: From a total of 143 patients studied, we found that 5.6% were heterozygous for the mutation at exon 7 (codon 306), and 7.7% were heterozygous for the mutation at exon 8 (codon 316). No homozygous subjects were found for either mutation. No significant correlation between these mutations and the presence of aPL, LAC, or anti-beta2GPI antibodies was found. In patients with SLE (n = 95), 4 of 6 patients with exon 8 mutation had thrombosis, compared with 22 of 82 patients without the mutation (P = 0.043). CONCLUSION: The prevalence of mutations in the fifth domain of beta2GPI in these patients with SLE and/or APS were similar to those previously reported for the general population. Heterozygosity for either mutation does not influence the incidence of aPL, but in patients with SLE, the mutation at exon 8 may predispose to thrombosis as an independent factor. PMID- 10366112 TI - High-dose versus low-dose D-penicillamine in early diffuse systemic sclerosis: analysis of a two-year, double-blind, randomized, controlled clinical trial. AB - OBJECTIVE: To test the hypothesis that systemic sclerosis (SSc) patients taking high-dose D-penicillamine (D-Pen) would have greater softening of skin, lower frequency of renal crisis, and better survival than patients taking low-dose D Pen. METHODS: Seventeen centers enrolled 134 SSc patients with early (< or =18 months) diffuse cutaneous scleroderma into a 2-year, double-blind, randomized comparison of high-dose D-Pen (750-1,000 mg/day) versus low-dose D-Pen (125 mg every other day). All 134 patients were followed up for a mean+/-SD of 4.0+/-1.1 years to assess the frequencies of new-onset scleroderma renal crisis (SRC) and mortality. RESULTS: Sixty-eight patients completed 24 months of drug treatment. The course of the modified Rodnan skin thickness score in the 32 high-dose and the 36 low-dose D-Pen completers was not different at 24 months: the skin score dropped 4.8+/-10.3 (mean+/-SD) units in the high-dose group and 6.9+/-8.4 units in the low-dose group (P = 0.384 by t-test; favoring low-dose D-Pen) from 20.4+/ 10.3 in the high-dose and 19.9+/-6.6 in the low-dose D-Pen group at study entry. The incidences of SRC and mortality were not different (P > 0.38 by Cox proportional hazards and by chi-square test) in the 66 high-dose patients (8 developed SRC and 8 died) compared with the 68 low-dose patients (10 developed SRC and 12 died). Of the 20 adverse event-related withdrawals, 80% occurred in the high-dose D-Pen group. CONCLUSION: The course of the skin score and the frequencies of SRC and mortality in the high-dose D-Pen group were not different from those in the low-dose D-Pen group. Eighty percent of the adverse event related withdrawals occurred in the high-dose D-Pen patients. Although this study cannot answer the question of whether low-dose D-Pen is effective, it does suggest that there is no advantage to using D-Pen in doses higher than 125 every other day. PMID- 10366114 TI - An assessment of the annual and long-term direct costs of rheumatoid arthritis: the impact of poor function and functional decline. AB - OBJECTIVE: To describe the distribution of direct medical care costs of rheumatoid arthritis (RA) over 1-year and 11-year periods, and to evaluate the impact of poor function and functional decline on direct costs. METHODS: The present study uses data from the University of California, San Francisco, RA Panel Study in which 1,156 persons with RA have been followed up for as long as 15 years through annual structured interviews and periodic updates on severity from rheumatologists. We present annual direct medical care cost data for the years 1995 and 1996 and estimates of cumulative costs for the period 1986-1996 for the 272 persons followed up continuously for this period. RESULTS: Medical care costs for RA averaged $5,919 a year from a societal perspective; persons with RA incur another $2,582 in medical care costs for non-RA reasons. Of the RA total costs, hospital admissions account for more than half. Costs are highly skewed, with the costs in the 90th, 95th, and 100th percentiles totaling $8,209, $31,059, and $85,469 a year, respectively. Cumulative costs for the period 1986 1996 averaged $57,201, with cumulative costs in the 90th, 95th, and 100th percentiles totaling $114,844, $142,563, and $191,540, respectively. Persons with RA in the worst quartile of function experienced total annual direct costs that were 2.55 times as high and total hospital costs that were 6.97 times as high as those in the best (e.g., the first) quartile. Poor baseline functional status and declining functional status had similar, large effects on cumulative medical care costs. CONCLUSION: Medical care costs for RA over 1 year and 1 decade are highly skewed. Persons with RA with poor and declining function experience much higher costs of care. PMID- 10366113 TI - Lack of efficacy of oral bovine type II collagen added to existing therapy in rheumatoid arthritis. AB - OBJECTIVE: To investigate the efficacy of oral type II collagen (CII) in the treatment of rheumatoid arthritis (RA), when added to existing therapy. METHODS: Patients with active RA (n = 190) were randomized into a 6-month, double-blind, placebo-controlled trial. Patients continued to take their current arthritis medications. Patients received either placebo or bovine CII, 0.1 mg/day for 1 month, then 0.5 mg/day for 5 months. RESULTS: There were no significant differences between the baseline characteristics of either group. The primary response parameter was the American College of Rheumatology (ACR) preliminary definition of improvement in RA (ACR 20). There was no statistically significant difference in the ACR 20 after 6 months (20.0% of placebo patients; 16.84% of bovine CII patients). There were significant differences in several clinical variables after treatment, all favoring the placebo group. CONCLUSION: Oral solubilized bovine CII, added to existing therapy, did not improve disease activity in patients with RA. PMID- 10366115 TI - Does rheumatoid arthritis remit during pregnancy and relapse postpartum? Results from a nationwide study in the United Kingdom performed prospectively from late pregnancy. AB - OBJECTIVE: To ascertain the influence of pregnancy on disease activity in women with rheumatoid arthritis (RA) during pregnancy and postpartum. METHODS: One hundred forty pregnant women were recruited from a nationwide campaign and were followed prospectively in the last trimester and at 1 and 6 months postpartum. Standardized assessment of joint symptoms, examination of inflamed joints, and the Health Assessment Questionnaire (HAQ) were the main measures of disease activity. RESULTS: There was only a modest fall in HAQ scores during pregnancy, with >25% of women having substantial levels of disability. Other parameters of disease activity showed a greater trend toward improvement, although only 23 (16%) were in complete remission (no joints with active disease and no therapy). Similarly, there was relatively little change in the distribution of HAQ scores from pregnancy to postpartum. There was, however, a statistically significant increase in the mean number of inflamed joints compared with the findings during pregnancy. Analysis of the possible influence of treatment suggested that therapy was associated with more severe disease and was not related to reduction in disease activity. CONCLUSION: This, the largest prospective study of the influence of pregnancy on RA activity, has demonstrated widespread variability in disease response. PMID- 10366116 TI - Excessive paternal transmission in psoriatic arthritis. AB - OBJECTIVE: The differential expression of a disease according to the sex of the disease-transmitting parent has been demonstrated in several autoimmune disorders. The purpose of the present study was to determine whether there are differences in the transmission and expression of psoriatic arthritis (PsA) that are dependent on the sex of the affected parent. METHODS: All probands (patients with PsA) were identified from among the patients attending the University of Toronto Psoriatic Arthritis Clinic. A self-reported family history of psoriasis or PsA was noted for each proband. Differences in parental and offspring transmission with respect to the proband were evaluated. In addition, the expression of PsA according to the sex of the affected parent was assessed at the time of the proband's presentation to the clinic. RESULTS: Ninety-five probands had affected parents: 62 (65%) had an affected father, and 33 (35%) had an affected mother. Thus, the proportion of paternal transmission (0.65) was significantly greater than was expected (0.5) (P = 0.001). Twelve of 74 offspring from male probands (16.2%) were affected with psoriasis or PsA, as compared with 9 of 108 offspring from female probands (8.3%) (P = 0.10). Probands whose fathers were affected had a higher frequency of skin lesions prior to arthritis (P = 0.047), an erythrocyte sedimentation rate > 15 mm/hour (P = 0.044), and a lower incidence of rheumatoid factor (P = 0.044). No differences were noted with respect to age at the onset of psoriasis or PsA, the severity of the PsA, or the frequency of HLA antigens. CONCLUSION: There appears to be excessive paternal transmission in PsA. Further clinical confirmation and elucidation of its genetic basis is warranted. PMID- 10366117 TI - Arthritis of the finger joints: a comprehensive approach comparing conventional radiography, scintigraphy, ultrasound, and contrast-enhanced magnetic resonance imaging. AB - OBJECTIVE: A prospective study was performed comparing conventional radiography, 3-phase bone scintigraphy, ultrasound, and magnetic resonance imaging (MRI) with precontrast and dynamic postcontrast examinations in 60 patients with various forms of arthritis including rheumatoid arthritis (RA), spondyl-arthropathy, and arthritis associated with connective tissue disease. METHODS: A total of 840 finger joints were examined clinically and by all 4 imaging methods. Experienced investigators blinded to the clinical findings and diagnoses analyzed all methods independently of each other. The patients were divided into 2 groups. Group 1 included 32 patients (448 finger joints) without radiologic signs of destructive arthritis (Larsen grades 0-1) of the evaluated hand and wrist and group 2 included 28 patients (392 finger joints) with radiographs revealing erosions (Larsen grade 2) of the evaluated hand and/or wrist. RESULTS: Clinical evaluation, scintigraphy, MRI, and ultrasound were each more sensitive than conventional radiography in detecting inflammatory soft tissue lesions as well as destructive joint processes in arthritis patients in group 1. All differences were statistically significant. We found ultrasound to be even more sensitive than MRI in the detection of synovitis. MRI detected erosions in 92 finger joints (20%; 26 patients) in group 1 that had not been detected by conventional radiography. CONCLUSION: Our data indicate that MRI and ultrasound are valuable diagnostic methods in patients with arthritis who have normal findings on radiologic evaluation. PMID- 10366118 TI - Larger increases in bone mineral density during alendronate therapy are associated with a lower risk of new vertebral fractures in women with postmenopausal osteoporosis. Fracture Intervention Trial Research Group. AB - OBJECTIVE: To investigate whether the incidence of vertebral fractures is related to the magnitude of change in bone mineral density (BMD) during alendronate treatment. METHODS: Women in this study were age 55-81 years (n = 2,984). While participating in the Fracture Intervention Trial, they received 5 mg/day of alendronate for 2 years followed by 10 mg/day for the remaining 12-30 months of the study. Their BMD was measured at baseline and at 12 and 24 months, and spine radiographs were obtained at baseline and again at 36 or 48 months to identify new vertebral fractures. RESULTS: After 12 months of alendronate treatment, 35% of participants had increases of > or =3% in total hip BMD, and 21% had either decreased total hip BMD or no change. Women who had larger increases in total hip BMD during the first 12 months had a lower incidence of new vertebral fractures during the entire followup period. Only 3.2% of women with increases of > or =3% in total hip BMD experienced new vertebral fractures, whereas twice as many women (6.3%) whose BMD declined or stayed the same experienced new fractures (adjusted odds ratio 0.45, 95% confidence interval 0.27-0.72). Similar patterns were observed for spine BMD at 12 months, and for both sites using change in BMD at 24 months. CONCLUSION: Women with increases of > or =3% in BMD during the first 1 or 2 years of alendronate treatment had the lowest incidence of new vertebral fractures. These findings suggest that, among women taking antiresorptive agents, greater increases in BMD are associated with lower risk of new vertebral fractures. PMID- 10366119 TI - The role of parvovirus B19 in the pathogenesis of giant cell arteritis: a preliminary evaluation. AB - OBJECTIVE: To determine whether parvovirus B19 DNA is more likely to be present in the temporal arteries of patients with giant cell arteritis (GCA) than in the temporal arteries of control subjects. METHODS: We prospectively examined temporal artery biopsy (TAB) tissue from 50 consecutive patients presenting for TAB for the presence of B19 DNA using the polymerase chain reaction (PCR). Clinical and demographic information was obtained from the patients' medical records. A separate PCR analysis of 30 original tissue specimens was conducted at the Centers for Disease Control and Prevention (CDC) using primers directed toward another target sequence in the nonstructural coding area of B19. RESULTS: The 50 patients had an average age of 70.8 years; 27 (54%) were female. Amplicons for human beta-globulin, but not for cytomegalovirus, were produced for all tissue samples. The PCR results for B19 agreed in 29 of 30 samples tested by our institution and by the CDC (97% agreement; kappa = 0.9). A comparison of the B19 DNA analysis and the results of TAB indicated a statistically significant association between histologic evidence of GCA and the presence of B19 DNA in TAB tissue (chi2 = 10.38, P = 0.0013). CONCLUSION: These findings suggest that B19 may play a role in the pathogenesis of GCA. PMID- 10366120 TI - Musculoskeletal manifestations in a population-based cohort of patients with giant cell arteritis. AB - OBJECTIVE: To define musculoskeletal manifestations occurring in a population based cohort of patients with giant cell (temporal) arteritis (GCA). METHODS: The records of 128 patients with GCA diagnosed over a 42-year-period (1950-1991) in Olmsted County, MN, were reviewed for the presence and type of musculoskeletal manifestations, their relationship to the onset and course of GCA, and their response to treatment. RESULTS: Fifty-three patients (41%) developed polymyalgia rheumatica: 23 before, 17 concurrently with, and 13 after the diagnosis of GCA. Thirty patients (23%) developed 1 or more peripheral musculoskeletal manifestations. These included peripheral synovitis in 23 patients (6 of whom fulfilled criteria for rheumatoid arthritis), distal extremity swelling with pitting edema in 13, distal swelling without pitting in 5, tenosynovitis in 6, and carpal tunnel syndrome in 2. Fifty-seven episodes of peripheral manifestations occurred in the 30 patients at different times during the course of GCA. In most, the onset of PMR and peripheral manifestations was within 2 years of the diagnosis of GCA. CONCLUSION: Musculoskeletal symptoms in GCA are common and varied. Most appear linked temporally to the underlying GCA, indicating that the nature of this illness and its clinical expression are broader than often considered. PMID- 10366121 TI - Reduced utilization and cost of primary care clinic visits resulting from self care education for patients with osteoarthritis of the knee. AB - OBJECTIVE: To determine the extent to which the cost of an effective self-care intervention for primary care patients with knee osteoarthritis (OA) was offset by savings resulting from reduced utilization of ambulatory medical services. METHODS: In an attention-controlled clinical trial, 211 patients with knee OA from the general medicine clinic of a municipal hospital were assigned arbitrarily to conditions of self-care education (group E) or attention control (group AC). Group E (n = 105) received individualized instruction and followup emphasizing nonpharmacologic management of joint pain. Group AC (n = 106) received a standard public education presentation and attention-controlling followup. A comprehensive clinical database provided data concerning utilization and cost of health services during the following year. RESULTS: Only 25 subjects (12%) were lost to followup. The 94 subjects remaining in group E made 528 primary care visits during the year following intervention, compared with 616 visits by the 92 patients remaining in group AC (median visits 5 versus 6, respectively; P < 0.05). Fewer visits translated directly into reduced clinic costs in group E, relative to controls (median costs [1996 dollars] $229 versus $305, respectively; P < 0.05). However, self-care education had no significant effects on utilization and costs of outpatient pharmacy, laboratory, or radiology services over the ensuing year. The cost per patient to deliver the self-care intervention was estimated to be $58.70. CONCLUSION: Eighty percent of the cost of delivering effective self-care education to the knee OA patients in this study was offset within 1 year by the reduced frequency and costs of primary care visits. For >50% of patients receiving the intervention, the savings associated with fewer primary care visits exceeded the cost of self-care education. PMID- 10366122 TI - Ovarian failure and flares of systemic lupus erythematosus. AB - OBJECTIVE: To study the effects of ovarian failure on disease flares in systemic lupus erythematosus (SLE). METHODS: Fifty-four female premenopausal SLE patients who were under the age of 45 years and treated with continuous oral cyclophosphamide (CYC) for no more than 12 months were studied. All patients had been followed up for >5 years following CYC treatment. Demographic characteristics, clinical and serologic profiles, and information concerning disease flares were recorded. Comparison of the number of severe and mild/moderate flares during the first 5 years after CYC treatment was made between patients who developed CYC-induced ovarian failure and those who did not. RESULTS: Fourteen SLE patients had documented ovarian failure with hypoestrogenemia within 2 years after CYC treatment. Compared with the menstruating group of patients, those who developed ovarian failure were significantly older at the time of CYC therapy (mean 37.9 versus 25.5 years; P < 0.001), but otherwise no significant differences in organ manifestations and autoantibody profiles between the 2 groups were observed. Both the ovarian failure group and menstruating group of patients had similar SLE Disease Activity Index scores at the time of CYC treatment (mean 15.6 versus 17.7; P = 0.16), and had comparable treatment durations (mean 8.2 versus 7.8 months; P = 0.68) and cumulative doses of CYC (mean 20.4 versus 17.9 grams; P = 0.34). Flares of SLE were uncommon during the first year following CYC administration. However, during the 5-year followup period, patients who developed CYC-induced ovarian failure had significantly fewer severe flares (mean 0.014 versus 0.075 flares/patient year; P = 0.01) and smaller total number of flares (mean 0.128 versus 0.250 flares/patient-year; P = 0.03) when compared with those who were still menstruating. CONCLUSION: This study provides an important clinical observation to support the notion that ovarian failure with hypoestrogenemia is protective against lupus flares and emphasizes the importance of estrogen status in the determination of disease activity in SLE. PMID- 10366123 TI - Chlamydia trachomatis nucleic acids can be found in the synovium of some asymptomatic subjects. AB - OBJECTIVE: The recent identification of antigens or nucleic acids of infectious agents in the joints of patients with reactive arthritis has raised questions about whether chlamydial or other infectious agent nucleic acids are also present in normal joints. We had the opportunity to study synovium from 30 asymptomatic volunteer subjects by use of polymerase chain reaction (PCR) for attempted identification of Chlamydia and other infectious agents. METHODS: All subjects had blind needle synovial biopsies with the Parker-Pearson needle. DNA was extracted and PCR performed using primers for Chlamydia trachomatis, Chlamydia pneumoniae, Borrelia burgdorferi, and pan bacterial 16S ribosomal RNA (rRNA). RESULTS: Two subjects were identified with nucleic acid for the 16S rRNA gene of C trachomatis. All other PCR reactions were negative except for the pan bacterial 16S rRNA in the C trachomatis-positive subjects. Both subjects, although symptom free, had some evidence of synovial reaction. CONCLUSION: C trachomatis appears to occasionally be disseminated to joints without producing overt disease. PMID- 10366124 TI - Genetic risk and protective factors for idiopathic inflammatory myopathy in Koreans and American whites: a tale of two loci. AB - OBJECTIVE: To better understand genetic contributions to autoimmunity, immunogenetic markers were studied in two racially discrete and geographically isolated populations of patients with idiopathic inflammatory myopathy (IIM). METHODS: Clinical characteristics, as well as clinical and autoantibody subsets, were defined in 151 American white patients and 50 Korean patients with IIM. HLA DRB1 and DQA1 genotyping was performed on patients and racially matched controls by standard molecular techniques. Gm allotypes and phenotypes were determined by the hemagglutination-inhibition method. RESULTS: HLA-DRB1*0301, the linked allele DQA1*0501, and DRB1 alleles sharing the first hypervariable region motif 9EYSTS13 were major genetic risk factors for the development of myositis in whites (corrected P [Pcorr] < 0.0004, odds ratio [OR] 11.2, 4.5, and 3.1, respectively, for each factor versus controls). Although both the white and Korean patients had a similar distribution of clinical characteristics, autoantibody profiles, and clinical groups, no HLA-DRB1 nor DQA1 allele or motif was found to be a risk factor for IIM in the Korean patients. However, DRB1*14 was a protective factor in Korean patients without myositis-specific autoantibodies (Pcorr = 0.004, OR 0.046). In addition, although no Gm phenotype or allotype was identified as a risk factor in whites, Gm 21 was a protective factor for the development of IIM in Koreans (Pcorr = 0.024, OR 0.3). CONCLUSION: Although myositis patients in the US and Korea share similar clinical and serologic features, the immune response genes predisposing to and protecting from myositis in each of these ethnic groups differ at two chromosomal loci. These data suggest that multiple genetic loci should be studied to identify risk and protective factors for some autoimmune diseases in various ethnic populations. PMID- 10366125 TI - An aggressive form of polyarticular arthritis in a man with CD154 mutation (X linked hyper-IgM syndrome). AB - Hyper-IgM syndrome (HIM) is a rare immunodeficiency disorder that has been associated with the development of symptoms and clinical features characteristic of rheumatoid arthritis (RA). We describe a patient with HIM and severe erosive arthritis with prominent nodules in the absence of detectable serum rheumatoid factor. Because HIM results from defects in either T cell CD154 (CD40 ligand) expression or abnormal CD40 signaling, the molecular basis of the patient's disease was analyzed. Activated CD4+ T cells failed to express surface CD154 protein, and molecular analysis of CD154 complementary DNA revealed a nucleotide transversion resulting in the nonconservative amino acid substitution G-D at amino acid 257. This case indicates that defective CD154-dependent CD40 signaling can be associated with susceptibility to a severe inflammatory arthritis that has both similarities to and differences from idiopathic RA. PMID- 10366126 TI - Clinical images: Skin necrosis in giant cell arteritis. PMID- 10366127 TI - Plasma adrenomedullin in rheumatoid arthritis compared with other rheumatic diseases. PMID- 10366128 TI - Association of interleukin-4 receptor and interleukin-4 promoter gene polymorphisms with systemic lupus erythematosus. PMID- 10366129 TI - Anti-[(H2A/2B)-DNA]] IgG supports the diagnosis of procainamide-induced arthritis or pleuritis. PMID- 10366130 TI - Recurrent acute calcium pyrophosphate dihydrate arthritis following intraarticular hyaluronate injection. PMID- 10366131 TI - Detecting peripheral nerve involvement in rheumatoid arthritis patients: comment on the article by Lanzillo et al. PMID- 10366132 TI - Autoantibodies against bactericidal/permeability-increasing protein in cystic fibrosis patients: comment on the article by Hoffman and Specks. PMID- 10366133 TI - Severe exacerbation of systemic lupus erythematosus after hepatitis B vaccination and importance of pneumococcal vaccination in patients with autosplenectomy: comment on the article by Battafarano et al. PMID- 10366134 TI - Illegal sales of cigarettes to minors--Ciudad Juarez, Mexico; El Paso, Texas; and Las Cruces, New Mexico, 1999. AB - In 1996, the United States-Mexico Binational Commission (US-MBC) Health Working Group identified prevention of tobacco use, particularly among adolescents, as a priority and subsequently recommended joint efforts toward reducing illegal sales of cigarettes to minors. A 1997 survey of 561 commercial cigarette outlets in Mexico City found that 79% of retailers sold cigarettes to minors. To assess the illegal sale of cigarettes to minors in other regions of Mexico and on both sides of the U.S.-Mexico border, during January-February 1999 the General Directorate of Epidemiology in Mexico, the Chihuahua State Department of Health Services (CDH), the Ciudad Juarez Department of Health (CJDH), the Texas Department of Health (TDH), and the New Mexico Department of Health (NMDH) surveyed cigarette outlets in Ciudad Juarez, Mexico; El Paso, Texas; and Las Cruces, New Mexico. This report summarizes the results of these surveys, which indicate that almost all retailers in the surveyed outlets in Ciudad Juarez sold cigarettes to minors and that sales rates to minors were substantially lower in El Paso and Las Cruces. PMID- 10366135 TI - Determination of nicotine, pH, and moisture content of six U.S. commercial moist snuff products--Florida, January-February 1999. AB - The use of smokeless tobacco (moist snuff and chewing tobacco) can cause oral cancer and precancerous oral lesions (leukoplakia) and is a risk factor for cardiovascular diseases and nicotine addiction. Despite these adverse effects, smokeless tobacco is used commonly in the United States by young people, especially male high school students. Officials in Florida requested CDC assistance in analyzing six moist snuff products to measure three factors that affect their nicotine dose: pH, nicotine content, and moisture content. This report summarizes the results of the analysis, which indicate that the pH, amount of nicotine, and moisture vary widely among brands. PMID- 10366136 TI - Prenatal discussion of HIV testing and maternal HIV testing--14 states, 1996 1997. AB - In July 1995, the Public Health Service recommended that health-care providers counsel all pregnant women about human immunodeficiency virus (HIV) prevention and encourage testing for HIV infection and, if indicated, initiate zidovudine therapy. To evaluate compliance with these recommendations, CDC analyzed population-based data on HIV counseling and testing during 1996-1997 from 14 states participating in the Pregnancy Risk Assessment Monitoring System (PRAMS). This report presents an analysis of survey data collected from 1996 through 1997; results indicate that HIV counseling and testing of pregnant women were common but varied by state, type of prenatal health-care provider, Medicaid status, and maternal demographic characteristics. PMID- 10366137 TI - Prevention of varicella. Update recommendations of the Advisory Committee on Immunization Practices (ACIP). AB - In February 1999, the Advisory Committee on Immunization Practices (ACIP) expanded recommendations for varicella (chickenpox) vaccine to promote wider use of the vaccine for susceptible children and adults. The updated recommendations include establishing child care and school entry requirements, use of the vaccine following exposure and for outbreak control, use of the vaccine for some children infected with the human immunodeficiency virus (HIV), and vaccination of adults and adolescents at high risk for exposure. These recommendations also provide new information on varicella vaccine postlicensure safety data. PMID- 10366138 TI - Prevention and control of influenza: recommendations of the Advisory Committee on Immunization Practices (ACIP). AB - This report updates 1998 recommendations by the Advisory Committee on Immunization Practices (ACIP) on the use of influenza vaccine and antiviral agents (MMWR 1998;47[No. RR-6]:1-26). The principal changes include a) information on the influenza virus strains included in the 1999-2000 trivalent vaccine; b) discussion of the potential expanded use of influenza vaccine; c) new background information on live-attenuated influenza vaccines (LAIVs), neuraminidase-inhibitor drugs, and rapid diagnostic tests; d) new information on the epidemiology of influenza among travelers; and e) the addition of referenced citations. This report and other information on influenza can be accessed at the website for the Influenza Branch, Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, CDC at . PMID- 10366139 TI - Knowledge and use of folic acid by women of childbearing age--United States, 1995 and 1998. AB - In the United States, approximately 4000 pregnancies are affected by neural tube defects each year; 50%-70% of these developmental defects could be prevented with daily intake of 400 microg of the B vitamin folic acid throughout the periconceptional period. In 1992, the Public Health Service recommended that all women capable of becoming pregnant consume 400 microg of folic acid daily throughout their childbearing years to reduce their risk for having a pregnancy affected by neural tube defects. In 1998, the Institute of Medicine recommended that all women of childbearing potential consume 400 microg of synthetic folic acid per day from fortified foods and/or a supplement in addition to food folate from a varied diet. This report summarizes the findings of a survey conducted during July-August 1998 to assess folic acid knowledge and practices among women of childbearing age in the United States and compares these results with those from a similar survey conducted in 1995. The findings indicate that 7% of women know folic acid should be taken before pregnancy to reduce the risk for neural tube defects. PMID- 10366140 TI - Outbreak of poliomyelitis--Angola, 1999. AB - On March 23, 1999, the Pediatric Hospital in Luanda, Angola, reported 21 cases (three deaths) of acute flaccid paralysis (AFP). By April 3, 102 AFP cases had been reported in Luanda and neighboring areas of Bengo province. A preliminary investigation by the Ministry of Health (MOH) indicated that these cases primarily occurred among children aged <5 years; 90% had received two or fewer doses of oral poliovirus vaccine (OPV), 4% had received three doses, and 6% had received four doses. Many case-patients resided in overcrowded municipalities where families displaced by civil war had settled. On the basis of preliminary data, MOH suspected the outbreak was poliomyelitis and began planning a vaccination campaign to control the epidemic. Surveillance was strengthened to identify and rapidly investigate reports of AFP cases to determine the extent of the outbreak. PMID- 10366141 TI - Playground safety--United States, 1998-1999. AB - Each year approximately 211,000 U.S. children receive emergency department care for injuries sustained on playground equipment, making the use of this equipment the leading cause of injuries to children in school and child care environments. In response to the problem, the National Program for Playground Safety (NPPS) at the University of Northern Iowa (UNI) developed a national action plan that focuses on four areas of playground injury prevention: supervision, age appropriateness of equipment, suitable fall surfaces, and equipment maintenance. During 1998-1999, NPPS surveyed a sample of the nation's child care, elementary school, and park playgrounds. This report summarizes the survey results, which indicate that playground injuries could be reduced by measures such as resilient surfacing below equipment, better equipment maintenance, improved supervision, and use of age-appropriate equipment. PMID- 10366142 TI - Childhood work-related agricultural fatalities--Minnesota, 1994-1997. AB - Agriculture is one of the most hazardous industries in the United States, with the second highest work-related fatality rate during 1992-1996 (21.9 deaths per 100,000 workers). During 1992-1995, 155 deaths were reported among agricultural workers aged < or =19 years; 64 (41%) of these youths were working in their family's business . In Minnesota during 1992-1996, agriculture had the highest fatality rate of any industry (21.3 per 100,000 workers). To characterize agriculture work-related deaths among youths in Minnesota during 1994-1997, the Minnesota Department of Health (MDH) analyzed data from the state's Fatality Assessment and Control Evaluation (FACE) program. This report presents five cases of agriculture work-related fatalities among youths in Minnesota. PMID- 10366143 TI - Update: outbreak of Nipah virus--Malaysia and Singapore, 1999. AB - During March 1999, health officials in Malaysia and Singapore, in collaboration with Australian researchers and CDC, investigated reports of febrile encephalitic and respiratory illnesses among workers who had exposure to pigs. A previously unrecognized paramyxovirus (formerly known as Hendra-like virus), now called Nipah virus, was implicated by laboratory testing in many of these cases. Febrile encephalitis continues to be reported in Malaysia but has decreased coincident with mass culling of pigs in outbreak areas. No new cases of febrile illness associated with Nipah virus infection have been identified in Singapore since March 19, 1999, when abattoirs were closed. This report summarizes interim findings from ongoing epidemiologic and laboratory investigations in Malaysia and Singapore. PMID- 10366144 TI - Evaluation of diagnostic criteria and behavior of ovarian intestinal-type mucinous tumors: atypical proliferative (borderline) tumors and intraepithelial, microinvasive, invasive, and metastatic carcinomas. AB - Histologic criteria for the distinction of ovarian mucinous borderline tumors (MBTs) from invasive mucinous carcinomas (MUCCAs) and the definitions of intraepithelial (noninvasive) carcinoma and microinvasion are controversial. Accurate assessment of the behavior of these tumors has been obscured by inclusion of cases of pseudomyxoma peritonei (PMP), an entity of extraovarian origin, and misclassification of the ovarian tumors in PMP and metastatic mucinous carcinomas (METCAs) as either advanced-stage MBTs or primary ovarian MUCCAs. One hundred thirty-six intestinal-type ovarian mucinous tumors without PMP were evaluated for the presence of stromal invasion, marked cytologic atypia, epithelial stratification of more than three cell layers, and necrosis. Criteria for the diagnosis of MBT, MBT with intraepithelial carcinoma, MBT with microinvasion (MIBT), MUCCA, and METCA were established and correlated with behavior. Twenty-three (59%) of 39 patients whose tumors had stromal invasion of more than 5 mm died of disease. Stromal invasion of more than 5 mm was the sole feature that correlated with a poor prognosis. In the absence of this feature, marked cytologic atypia, epithelial stratification of more than three layers, microinvasion (<5 mm), or necrosis did not have an adverse effect on survival. Tumors were classified as MBT (n = 65; 48%) based on absence of stromal invasion, regardless of degree of cytologic atypia or epithelial stratification; of these, 28 (43%) qualified as intraepithelial carcinoma based on epithelial stratification of more than three cell layers or marked cytologic atypia. Tumors with stromal invasion of less than 5 mm were classified as MIBT (n = 8; 6%). Tumors with stromal invasion of more than 5 mm were classified as MUCCA (n = 24; 18%). Tumors with a nodular pattern of stromal invasion, morphology inconsistent with ovarian origin, or a primary site elsewhere were classified as METCA (n = 35; 26%). Four tumors could not be definitively classified. Of the 86 cases with follow-up (median, 33 months) all MBTs (n = 44) and MIBTs (n = 6) were stage I, with 5-year survival rates of 100%. MUCCAs (n = 17) that were stage I had a 5 year survival rate of 91%; all patients with advanced-stage MUCCA died of disease. Five-year survival rate for METCAs (n = 19) was 11%. METCAs were more common than MUCCAs but can mimic MUCCAs and MBTs clinically and histologically. In the absence of a primary site elsewhere, METCA should be strongly suspected when there is bilateral surface involvement and a characteristic nodular pattern of invasion. It is important to recognize this pattern because 5-year survival rate for METCA (11%) was substantially less than that of MUCCA (all stages, 51%) and MBT (100%). Because all MBTs, regardless of degree of atypia or stratification, were stage I and benign, we prefer to designate them as atypical proliferative mucinous tumors. Marked cytologic atypia, epithelial stratification of more than three layers, and microinvasion (<5 mm) had no effect on the behavior of MBT. Noninvasive mucinous tumors with marked cytologic atypia or excessive epithelial stratification can be interpreted as atypical proliferative tumors with intraepithelial carcinoma and those with microinvasion can be designated as atypical proliferative tumors with microinvasion; these tumors appear to represent transitional stages in ovarian mucinous carcinogenesis. PMID- 10366145 TI - Extraskeletal myxoid chondrosarcoma: a reappraisal of its morphologic spectrum and prognostic factors based on 117 cases. AB - Extraskeletal myxoid chondrosarcoma (EMC), a phenotypically and genotypically distinctive entity, has generally been viewed as a low-grade sarcoma. No studies regarding clinical and morphologic prognostic factors have been performed on a large series of cases with long-term follow-up because of the rarity and protracted clinical course of EMC. The clinical, morphologic, and immunohistochemical features of 117 previously unreported cases were studied and statistically analyzed. The male-to-female ratio was 2:1. The median patient age was 52 years (range, 6-89 years), and the median tumor size was 7 cm (range, 1.1 25 cm). All tumors occurred within the deep subcutis or deeper soft tissues, with 80% occurring in the proximal extremities or limb girdles and 20% in the trunk. Most initial tumor excisions were intralesional or marginal. Follow-up information was available in 99 cases (median, 9 years: range, 2 months-22 years). Forty-eight patients were disease-free, and 41 patients had evidence of disease (18 of these had died of disease). Ten additional patients survived, but their disease status was unknown. There were local recurrences in 40 (48%) of 83 patients, 23 (58%) of whom had multiple local recurrences. Metastases occurred in 35 (46%) of 76 patients. The estimated 5-, 10-, and 15-year survival rates were 90%, 70%, and 60%, respectively. All cases had histologic features characteristic of classical EMC, at least focally. Cellular foci devoid of myxoid matrix and reminiscent of chondroblastoma, Ewing's sarcoma, monophasic and poorly differentiated synovial sarcoma, fibrosarcoma, and rhabdoid tumor were identified in 29% cases. Older patient age, larger tumor size, and tumor location in the proximal extremity or limb girdle were adverse prognostic factors identified by multivariate analysis. Metastasis also adversely affected survival, although local recurrence did not. This study shows that EMC has a unique clinical course, including a high rate of local recurrence, prolonged survival after metastasis in some cases, and eventually a high rate of death due to tumor. These features distinguish EMC from low-grade sarcomas. This study shows that histologic grading is of no prognostic value in EMC because prognosis is dictated primarily by certain clinical features. Histologic recognition of classical EMC and cellular and solid, nonmyxoid variants is important, however, in view of EMC's distinctive biologic behavior. PMID- 10366146 TI - Intestinal adenocarcinoma in Crohn's disease: a report of 30 cases with a focus on coexisting dysplasia. AB - There are relatively few reports that detail the types of intestinal adenocarcinoma complicating Crohn's disease and examine associated epithelial dysplasia. We determined the prevalence, grade, and type of dysplasia found adjacent to and distant from Crohn's-related adenocarcinomas. Thirty cases of resected Crohn's-related adenocarcinoma were reviewed, and histologic type, degree of differentiation, TNM stage, and the presence or absence, grade, and location of dysplasia were recorded. Most of the patients were male (70%). The median ages at diagnosis of Crohn's disease and adenocarcinoma were 34 and 49 years, respectively. The extent of Crohn's disease included ileocolitis in 21 patients, only colonic disease in six, and only small bowel disease in three. In most cases (67%), carcinoma was found incidentally at surgery. All carcinomas arose in areas involved by Crohn's disease. Eight (27%) adenocarcinomas arose in the small bowel, and 22 (73%) arose in the colon, including two in out-of-circuit rectums. Most carcinomas (63%) were poorly differentiated. Dysplasia was found adjacent to the carcinoma in 26 (87%) cases. Of the colorectal carcinomas, 19 (86%) had adjacent dysplasia, and nine (41%) had distant dysplasia. In conclusion, most cases of Crohn's-related intestinal adenocarcinoma have dysplasia in adjacent mucosa, and 41% of those arising in the colorectum have distant dysplasia, supporting a dysplasia-carcinoma sequence in Crohn's disease. PMID- 10366147 TI - Spindle cell tumors associated with mycobacteria in lymph nodes of HIV-positive patients: 'Kaposi sarcoma with mycobacteria' and 'mycobacterial pseudotumor'. AB - Patients infected with HIV often have unusual manifestations of common infections and neoplasms. One such example is "mycobacterial pseudotumor," an exuberant spindle cell lesion induced in lymph nodes by mycobacteria. Kaposi sarcoma also produces a spindle cell proliferation in lymph nodes of HIV-positive patients. These two entities must be differentiated from one another because of differences in treatment and prognosis. We report here, however, three cases of intranodal Kaposi sarcoma with simultaneous mycobacterial infection, the occurrence of which has not been clearly documented. For comparison, we also studied three cases of mycobacterial pseudotumor, of which 14 cases have been described to date. There was considerable histologic overlap between these two lesions. Acid-fast bacilli were present in all cases, predominantly in the more epithelioid histiocytes in the cases of Kaposi sarcoma, and in spindle and epithelioid cells in the cases of mycobacterial pseudotumor. The morphologic features that favored Kaposi sarcoma over mycobacterial pseudotumor were the prominent fascicular arrangement of spindle cells and slitlike spaces, the lack of granular, acidophilic cytoplasm, and the presence of mitoses. Immunohistochemistry was a reliable adjunct study in the differential diagnosis, as the spindle cells in mycobacterial pseudotumor were positive for S-100 protein and CD68 whereas those of Kaposi sarcoma were CD31- and CD34-positive but negative for S-100 protein and CD68. Awareness that Kaposi sarcoma may coexist with mycobacterial infection in the same biopsy specimen is important because these lesions may be misdiagnosed as mycobacterial pseudotumor. The clinical impact of distinguishing between Kaposi sarcoma with mycobacteria and mycobacterial pseudotumor is significant because the presence of Kaposi sarcoma alters treatment and prognosis. PMID- 10366148 TI - Plexiform fibrohistiocytic tumor: clinicopathologic analysis of 22 cases. AB - Twenty-two cases of plexiform fibrohistiocytic tumor were reviewed to perform a clinicopathologic correlation with the behavior of the neoplastic entity. The tumor arises more frequently in children, adolescents, and young adults (mean age of presentation, 14.6 years), with strong female predilection (F:M, 6:1). It involves preferentially the upper extremity (64%), especially the fingers, hand, or wrist (45%). Most patients present with a small (average size, 2.5 cm; range, 0.5-8 cm) painless mass that slowly enlarges for months to years. All tumors involve subcutaneous adipose tissue, with extension into the dermis (19%), skeletal muscle (14%), or both (14%). Grossly, the tumors characteristically are poorly circumscribed and of firm consistency. Histologically, they are characterized by a plexiform proliferation of mononuclear histiocyte-like cells, multinucleated osteoclast-like cells, and spindle fibroblast-like cells in variable proportions and have three distinct growth patterns: fibrohistiocytic (36% of tumors), fibroblastic (32%), and mixed (32%), depending on the predominant cell type. Cellular atypia and pleomorphism are usually absent or minimal. Most tumors (78%) display mitotic activity, frequently <3 mitoses/10 high-power fields, and only 14% of the lesions display atypical mitoses. Vascular invasion was seen in only one tumor. Immunohistochemically, all tumors evaluated reacted with antibodies to CD68 that stained mainly the multinucleated giant cells and, to a lesser extent, mononuclear histiocyte-like cells and, occasionally, fibroblast-like cells. Less frequently, staining with antiactin antibodies was observed, restricted mainly to spindle cells. All nine tumors examined had a diploid DNA content. According to latest follow-up data (average period, 3.6 years) from 16 patients, 13 (82%) were alive with no evidence of disease (average, 3.6 years), 1 (6%) was alive with metastatic disease (follow up, 2.3 years), 1 (6%) was alive with a stable pulmonary nodule of unknown nature (follow-up, 1.75 years), and 1 (6%) had died of disease 3 years after local recurrence and regional lymph node and pulmonary metastases developed. Two patients (12.5%) had local recurrence, 1 (6%) had regional lymph node metastasis, and 3 (19%) had pulmonary metastases. No proven association between clinicopathologic features and outcome was identified. In conclusion, plexiform fibrohistiocytic tumor is a rare mesenchymal neoplasm of young persons characterized by low-grade malignant behavior and is prone to recur locally and occasionally to metastasize regionally and systemically. PMID- 10366149 TI - Ciliated hepatic foregut cyst: a study of six cases and review of the literature. AB - Ciliated hepatic foregut cyst (CHFC) is a rare, benign, solitary cyst consisting of ciliated pseudostratified columnar epithelium, subepithelial connective tissue, a smooth muscle layer, and an outer fibrous capsule. We studied six previously unreported cases of CHFC and 50 cases from the literature. The literature search revealed that Friedreich first described the lesion in 1857 and hypothesized its congenital origin. The cyst generally is found incidentally on radiologic imaging or during surgical exploration, although one case presented with portal vein compression. It occurs more frequently in men and is found most commonly in the medial segment of the left hepatic lobe, unlike most other solitary cysts that show a female predominance and greater occurrence in the right hepatic lobe. Two of the 56 cases were multilocular. There has been an increase in the number of reports of CHFC during the past 15 years. This may reflect the increased availability and use of various radiologic imaging modalities. A large number of cases have been reported in the Japanese population, but the significance of this is unclear. CHFC should be considered in the differential diagnosis of other solitary liver cysts, including simple cysts, hepatobiliary cystadenomas, and parasitic cysts. PMID- 10366150 TI - Downregulation of p27KIP1 and Ki67/Mib1 labeling index support the classification of thyroid carcinoma into prognostically relevant categories. AB - The cyclin-dependent kinase inhibitor p27KIP1 has been proposed as a valuable prognostic indicator for a variety of human neoplasms. Immunohistochemical reactivity for p27KIP1 and the proliferation marker Ki67/Mib1 were investigated in 90 thyroid carcinomas of follicular cell origin. The neoplasms were divided into three prognostic groups on the basis of their morphologic features: group 1, well-differentiated papillary or follicular carcinomas with favorable pathologic features (43 papillary carcinomas and 4 minimally invasive follicular carcinomas); group 2, papillary or follicular carcinomas with unfavorable pathologic features (21 poorly differentiated carcinomas and 2 papillary carcinomas, tall cell variant); and group 3, undifferentiated, or anaplastic, carcinomas. p27KIP1 expression (p = 0.007) and Ki67/Mib1 labeling index (p = 0.02) showed a strong correlation with the subdivision of the thyroid carcinomas in the three prognostic groups with a significant linear trend for tumors with low p27KIP1 (p = 0.002) and high Ki67/Mib1 labeling index (p = 0.005) to segregate into the unfavorable categories (groups 2 and 3). Low p27KIP1 expression, but not cellular proliferation, was related to adverse prognostic factors, such as large tumor size (p = 0.03) and extrathyroidal extension (p = 0.01), but the correlation was not independent of the subdivision in the three groups. Low p27KIP1 expression (p = 0.03) and high proliferative rate (p = 0.02) were associated with poor survival, reflecting the close association between patient morbidity and mortality rates and tumor differentiation. No significant association could be seen between p27KIP1 or cellular proliferation and clinicopathologic parameters (e.g., age, sex, tumor size, extrathyroidal extension, vascular invasion, lymph node metastases, distant metastases, tumor stage, and survival rate) within any of the groups, or the histologic diagnosis of papillary versus follicular carcinoma irrespective of their degree of differentiation. Modulation of p27KIP1 and cellular proliferation patterns in thyroid carcinoma correlate with tumor differentiation and support the morphologic classification of thyroid carcinoma into prognostically relevant categories. PMID- 10366151 TI - Pathologic examination of the sentinel lymph node in malignant melanoma. AB - Sentinel lymphadenectomy is gaining increasing popularity in the staging and treatment of patients with melanoma at risk for metastases. As a result, pathologists are encountering these specimens more frequently in their daily practice. The pathologic status of the sentinel lymph node is pivotal to the patient's care because it provides staging information that dictates the need for further therapy, and therefore detailed pathologic assessment is warranted. A standard pathology protocol to handle these nodes has been developed at our institution and involves complete submission of all tissue with routine use of immunohistochemical staining for S-100 protein. By using this protocol, 838 sentinel lymph nodes from 357 patients have been examined, and metastases were found in 16% of patients. Although the metastasis was clearly seen on sections stained with hematoxylin and eosin in 55% of the positive patients, the immunostain showed metastatic disease not appreciable on initial hematoxylin and eosin screening in an additional 28 lymph nodes (45% of node-positive patients). Intraoperative touch preparation cytology may be used as an adjunct technique in sentinel lymph nodes grossly suspicious for metastatic disease. This technique has been performed on 23 sentinel lymph nodes, with no false positives and an overall sensitivity of 62%. The thorough pathologic evaluation of sentinel lymph nodes in patients with malignant melanoma requires complete submission of all tissue, routine use of immunohistochemistry, and touch preparation cytology in selected cases. PMID- 10366152 TI - Spindle cell thymic carcinoma: clinicopathologic and immunohistochemical study of a distinctive variant of primary thymic epithelial neoplasm. AB - We report 16 cases of a distinctive variant of primary thymic epithelial neoplasm characterized by prominent spindling of the tumor cells. The patients were seven women and nine men aged 23 to 82 years (mean, 54 years). The lesions presented as anterior mediastinal masses without clinical or radiographic evidence of tumor elsewhere. Most patients had chest pain, dyspnea, and cough; in five patients, the tumors were asymptomatic and were discovered on routine clinical examination. Grossly, the lesions were firm, well-circumscribed, and locally infiltrative, and had a firm cut surface with foci of hemorrhage, necrosis, and cystic changes. Most of the tumors were treated by complete surgical excision. Histologically, they were characterized by a spindle cell proliferation showing varying degrees of atypia and mitotic activity. In 12 cases, transitions could be seen with areas that showed the features of conventional spindle cell thymoma. In two cases, areas showing features of poorly differentiated (lymphoepitheliomalike) carcinoma and anaplastic carcinoma could also be observed. Immunohistochemical studies in 10 cases showed strong positivity of the spindle tumor cells for CAM5.2 cytokeratin, and negative staining for a panel of antibodies including epithelial membrane antigen, carcinoembryonic antigen, actin, desmin, vimentin, S-100 protein, HMB45, CD34, CD5, and CD99. Clinical follow-up of eight patients showed an aggressive biologic behavior with recurrence, metastasis, and death by tumor in five of them 2 to 5 years after diagnosis. Based on these findings, the present tumors are interpreted as an unusual spindle cell variant of thymic carcinoma. The close association of these cases with areas showing the features of spindle cell thymoma within the same tumor mass suggests that some of these lesions may arise as a result of malignant transformation in a preexisting spindle cell thymoma. PMID- 10366153 TI - Selective distribution of calretinin in adenocarcinomas of the human colon and adjacent tissues. AB - The expression of calretinin, a calcium-binding protein, has been studied in a series of 82 human colorectal adenocarcinomas. In 22.5% of the cases, part of the tumor cells were calretinin-positive, whereas the cells of the normal and paratumoral mucosa were always negative. Two types of cells from the tumoral mass reacted positively and selectively with calretinin-antisera: the tumor cells and giant fibroblasts. The neurons of enteric ganglia and reactive mesothelial cells also reacted positively to the same antibody. The results obtained by immunochemistry have been confirmed by Western blot analysis and in situ hybridization for calretinin mRNA. There is a correlation between the expression of calretinin and the degree of differentiation of the tumor. Well-differentiated tumors express calretinin in only 5% of the cases, whereas this percentage is 20% for moderately differentiated tumors and 66.6% for poorly differentiated or undifferentiated tumors. We conclude that calretinin is expressed by most undifferentiated colorectal adenocarcinomas, but only by a limited number of cells in well-differentiated tumors. The degree of its expression coincides also with additional signs of malignancy, such as an increase in the number of metastases in the regional lymph nodes and in other organs. PMID- 10366154 TI - Spindle epithelial tumor with thymus-like differentiation (SETTLE) of the thyroid with prominent mitotic activity and focal necrosis. AB - Spindle epithelial tumor with thymus-like differentiation (SETTLE) is a rare, apparently low-grade spindle cell tumor of the thyroid gland occurring in young individuals and thought to be derived from thymic or branchial pouch remnants. Spindle epithelial tumor with thymus-like differentiation has little to no mitotic activity, and focal necrosis has been reported in one case. We present a case of SETTLE in a 29-year-old man that was initially evaluated by fine-needle aspirate biopsy and ultimately found to be consistent histologically with SETTLE. In this case, there were numerous mitotic figures among the spindle cells and focal necrosis. Spindle epithelial tumor with thymus-like differentiation has been considered to be a tumor of low malignant potential with metastases developing some years after diagnosis. This is the first case in which prominent mitotic activity and necrosis is reported perhaps representing an aggressive variant. PMID- 10366156 TI - Primary pulmonary malignant meningioma. AB - Fewer than 20 cases of primary pulmonary meningioma have been reported. Most of these cases have been histologically and clinically benign. We report an unusual case of primary pulmonary malignant meningioma with atypical histologic features and malignant behavior. A computed tomography scan of the head did not show evidence of tumor. The right upper lobe mass was resected and showed features of an atypical meningioma with loss of architectural pattern, mild nuclear pleomorphism, increased mitotic counts (up to 15 mitotic figures per 10 high power fields), and focally prominent nucleoli. Focally, cells with rhabdoid features were identified. The tumor's immunohistochemical and ultrastructural profiles were consistent with a meningioma. The tumor stained negative for estrogen and focally positive for progesterone receptors and had a MIB-1 labeling index (marker of cell proliferation) of 9.2%. Approximately 5 months after the initial resection, the patient experienced a tumor recurrence with multiple lymph node metastases, spread to the middle and lower lobes of the right lung, and metastasis to the diaphragm. Rarely, primary pulmonary meningiomas may present as high-grade malignant lesions. PMID- 10366155 TI - Beta-HCG aberrant expression in primary mediastinal large B-cell lymphoma. AB - We report on a primary mediastinal large B-cell lymphoma with aberrant expression of beta-human chorionic gonadotropin (beta-hCG). The patient, a 33-year-old man, had cough, dyspnea, fever, superior vena cava syndrome, and a mediastinal bulky tumor. A biopsy showed that the latter was characterized by large cells, sclerosis, and compartmentalization. The neoplastic elements expressed CD45, CD20, CD79a and, partially, CD30, whereas they were negative for CD3, epithelial membrane antigen and cytokeratins. Surprisingly, they displayed a clear-cut positivity for beta-hCG. The remaining oncofetal markers applied (PLAP and alpha1 fetoprotein) were negative. Electron microscopy demonstrated the presence of numerous nuclear pockets and the lack of intercellular junctions. DNA analysis by polymerase chain reaction showed clonal rearrangement of Ig heavy-chain genes. The patient responded promptly to the administration of MACOP-B. To the best of our knowledge, this is the first example of B-cell lymphoma showing positivity for beta-hCG; a similar aberrant expression was previously observed only in three Japanese patients with human T-cell lymphotropic virus type I+ adult T-cell lymphoma/leukemia. Because primary mediastinal large B-cell lymphoma has in the past been frequently confused with germ cell tumors, pathologists should be aware of possible beta-hCG expression by lymphomatous cells to avoid the risk of misdiagnosis. PMID- 10366157 TI - Metanephric adenofibroma: report of a case and review of the literature. AB - The recent recognition of a variety of pediatric renal tumors of different biologic behavior places an ever-increasing demand on the surgical pathologist for an accurate diagnosis. Although metanephric adenofibroma is one of the rarest benign renal tumors, the clinical importance of correctly diagnosing it cannot be overemphasized because it can potentially be mistaken as Wilms' tumor. We describe the clinical, radiologic, and pathologic features of a case of metanephric adenofibroma and discuss its differential diagnosis. The neoplasm was composed of two discrete components: a major fibroblastic element and a minor immature epithelial element. The latter formed a small nodule beneath the renal capsule, which could barely be detected by magnetic resonance imaging. This subcapsular nodule, however, was slightly soft and tan and was distinctly different from the white, whorled cut surface of the main tumor. It was formed by closely packed small immature epithelial cells in a slightly edematous background, which was histologically identical to metanephric adenoma and closely resembled epithelial Wilms' tumor. Unlike Wilms' tumor, however, the epithelial cells were very bland with no mitoses. The main bulk of the tumor was formed by spindle fibroblastic cells that were cytologically similar to the spindle cells in congenital mesoblastic nephroma. The tumor, however, was well demarcated without the irregular infiltrating edges of congenital mesoblastic nephroma. In contrast to the randomly distributed epithelial element throughout the stromal component in previous reported cases of metanephric adenofibroma, our finding of the exceedingly small and discrete epithelial component expands the known histologic spectrum of the disease. In addition, the presence of such minute epithelial nodule underscores the importance of diligent pathologic examination and careful sampling of tissue for histologic examination. PMID- 10366158 TI - Recurring fibro-obliterative venopathy in liver allografts. AB - Recurrent diseases in liver allografts are not uncommon. These occur most frequently in those transplanted for viral hepatitis B and C. We report an unusual case of recurrent process in two consecutive liver allografts received by a 37-year-old woman, who previously had an unremarkable past medical history but developed a rapidly progressive cholestatic liver failure. Histopathologic examination of the native liver showed fibroocclusive lesions of both terminal hepatic venules and portal vein branches. The exuberant fibroobliterative process created dense fibrosis with whorled appearance, and broad fibrous septa connecting adjacent central areas, and sometimes bridging portal to central areas. Dense portal fibrosis resulted in compression atrophy and loss of bile ducts. The first allograft, which failed within 3 months, showed histopathologic findings similar to that of the native liver. A liver biopsy that was performed 20 months after the second liver transplant again showed similar histopathology. The histopathologic features and clinical presentation of this patient suggest an unusual form of recurring progressive fibroobliterative venopathy causing liver failure. PMID- 10366159 TI - Thymoma and thymic carcinoma. PMID- 10366160 TI - Serum IL-6 level and the development of disability in older persons. AB - BACKGROUND: The serum concentration of interleukin 6 (IL-6), a cytokine that plays a central role in inflammation, increases with age. Because inflammation is a component of many age-associated chronic diseases, which often cause disability, high circulating levels of IL-6 may contribute to functional decline in old age. We tested the hypothesis that high levels of IL-6 predict future disability in older persons who are not disabled. METHODS: Participants at the sixth annual follow-up of the Iowa site of the Established Populations for Epidemiologic Studies of the Elderly aged 71 years or older were considered eligible for this study if they had no disability in regard to mobility or in selected activities of daily living (ADL), and they were re-interviewed 4 years later. Incident cases of mobility-disability and of ADL-disability were identified based on responses at the follow-up interview. Measures of IL-6 were obtained from specimens collected at baseline from the 283 participants who developed any disability and from 350 participants selected randomly (46.9%) from those who continued to be non-disabled. FINDINGS: Participants in the highest IL 6 tertile were 1.76 (95% CI, 1.17-2.64) times more likely to develop at least mobility-disability and 1.62 (95% CI, 1.02-2.60) times more likely to develop mobility plus ADL-disability compared with to the lowest IL-6 tertile. The strength of this association was almost unchanged after adjusting for multiple confounders. The increased risk of mobility-disability over the full spectrum of IL-6 concentration was nonlinear, with the risk rising rapidly beyond plasma levels of 2.5 pg/mL. INTERPRETATION: Higher circulating levels of IL-6 predict disability onset in older persons. This may be attributable to a direct effect of IL-6 on muscle atrophy and/or to the pathophysiologic role played by IL-6 in specific diseases. PMID- 10366161 TI - The importance of subsyndromal depression in older primary care patients: prevalence and associated functional disability. AB - OBJECTIVE: Existing diagnostic categories for depression may not encompass the majority of older people suffering clinically significant depressive symptoms. We have described the prevalence of subsyndromal depressive symptoms and tested the hypothesis that patients with subsyndromal depression have greater functional disability and general medical burden than nondepressed subjects but less than patients with diagnosable depressions. METHODS: Subjects were 224 patients, aged 60 years and older, recruited from private internal medicine offices or a family medicine clinic. Validated measures of psychopathology, medical burden, and functional status were used. The subsyndromal depression group was defined by a score of more than 10 on the Hamilton Rating Scale for Depression and by the absence of major or minor depressive disorder. Analyses included multiple regression techniques to determine the presence of group differences adjusted for demographic covariates. RESULTS: Subsyndromal depression was common (estimated point prevalence of 9.9% compared with 6.5% for major depression, 5.2% for minor depression, and .9% for dysthymic disorder), associated with functional disability and medical comorbidity to a degree similar to major or minor depression, and often treated with antidepressant medications. CONCLUSIONS: Although depressive conditions are common and are associated with considerable functional and medical morbidity in older primary care patients, many patients with clinically significant depressive symptoms are not captured by criteria based syndromic diagnostic categories. Future work should include intervention studies of subsyndromally depressed older persons as well as attention to the course and biopsychosocial concomitants of diagnosable and subsyndromal depressions in this population. PMID- 10366162 TI - Higher systemic arterial compliance is associated with greater exercise time and lower blood pressure in a young older population. AB - OBJECTIVES: Arterial compliance is an important therapeutic target in older individuals in whom stiffening of the proximal arterial circulation is thought to underlie systolic hypertension and increased cardiac work. We have shown previously that arterial compliance is related to aerobic fitness and that it is increased in young (20 to 35 years old), previously sedentary individuals by a 4 week period of moderate aerobic training. The extent to which compliance relates to exercise performance in a random selection of young older patients has not been reported previously. Therefore, we examined the interrelationship between systemic arterial compliance (SAC) and time to cessation of exercise during a standard treadmill exercise test in an older population. DESIGN: A cross sectional survey. METHODS: SAC was estimated at rest using simultaneous recordings of ascending aortic flow and carotid applanation tonometry in 43 subjects aged 67 +/- 7 years (mean +/- SD; 24 men and 19 women). Treadmill exercise testing was performed using a modified Bruce protocol. Aerobic capacity was assessed as Heart Rate-Blood Pressure product and exercise tolerance as total treadmill time. RESULTS: SAC and exercise tolerance were related to gender, with men exhibiting greater exercise reserve and higher SAC than women. There was a significant positive correlation between SAC and time to cessation of exercise (r = .34; P = .03), with a negative correlation between SAC and resting heart rate blood pressure product (r = -.66; P < .001). SAC was correlated with height and blood pressure. Exercise tolerance was related to height (P < .02). CONCLUSIONS: These data indicate a positive association between SAC and fitness level in healthy older people and an inverse association between SAC and systolic blood pressure. Our findings are consistent with either (1) acquisition of a more compliant circulation and lower blood pressure through more physical activity or (2) that a more compliant arterial circulation and lower blood pressure permit greater athletic performance. PMID- 10366163 TI - Vascular compliance as a measure of biological age. AB - OBJECTIVES: To determine the measure of vascular compliance most closely related to age. DESIGN: A review of 22 studies relating aortic compliance to age and a discussion of other factors related to vascular compliance. MEASUREMENTS: Aortic compliance, elastic modulus, postmortem aortic changes, pulse wave velocity in the aorta, common carotid, lower limb and upper limb. RESULTS: 1. Aortic compliance and carotid artery compliance is closely related to age; 2. Compliance in the peripheral arteries, in 16 reports, appears less closely related to age; 3. There is evidence that aortic compliance is related to hypertension, cardiac function, and left ventricular hypertrophy and can be increased by exercise, hormonal therapy, antioxidant and antihypertensive treatment; and 4. Vascular compliance is more closely related to chronological age than other measures such as skin inelasticity, greying of hair, baldness etc. CONCLUSIONS: Because of the close relationship between aortic and carotid compliance and chronological age, deviation from the age-predicted norm (biological age) may prove to be a good predictor of cardiovascular pathology. PMID- 10366164 TI - Heart failure in community-dwelling older persons: aims, design and adherence rate of the ICARe Dicomano project: an epidemiologic study. Insufficienza Cardiaca negli Anziani Residenti a Dicomano. AB - BACKGROUND: The prevalence of heart failure (HF) increases with age, and HF is a major cause of disability and mortality in older persons. Detection of HF in epidemiological studies has relied on criteria validated only in young and middle age adults, and, therefore, may prove inadequate in older subjects, because they do not take into account the pathophysiologic and clinical peculiarities of HF in old age. Thus, the true prevalence of HF in the older general population remains uncertain and has probably been underestimated in previous studies. Moreover, the mechanism and the extent by which HF hinders physical functioning in older people has not been fully elucidated. OBJECTIVES: This paper describes the design of the ICARe study, carried out in an older home-dwelling population to collect data about: (1) the sensitivity and specificity of diagnostic criteria used previously in epidemiological studies of HF; (2) the prevalence of the different pathophysiologic forms of HF; and (3) the impact of HF on overall health status, and on physical functioning, in the absence or presence of chronic comorbidity. DESIGN AND SETTING: This was a cross-sectional survey. Eligible were all community-dwelling persons aged 65 years or older recorded in the Registry Office of Dicomano, a small town nearby Florence (Italy). All the domains of multidimensional geriatric assessment were explored through different phases of the study (home interview, laboratory testing, geriatric visit) that comprised an extensive cardiopulmonary instrumental assessment including: color Doppler echocardiography, echotomography of the carotid arteries used in an original method to determine arterial compliance, and bell spirometry. Presence of major chronic conditions was ascertained by predefined, standard algorithms that were based largely on clinical examination. RESULTS: There were 864 older persons eligible for the ICARe study. Even with a substantial decline from home interview (91.2%) to the cardiopulmonary study (71.1%), the adherence rate remained high throughout the study, and the population examined was fairly representative of the original eligible population. Thus, we believe that the data collected in this study offer a unique opportunity to assess the validity of the diagnostic clinical criteria for HF in the general older population, to identify the pathophysiology underlying the syndrome, and to investigate the relationship between HF, comorbidity, and disability. PMID- 10366165 TI - Who walks? Factors associated with walking behavior in disabled older women with and without self-reported walking difficulty. AB - OBJECTIVES: To determine how severity of walking difficulty and sociodemographic, psychosocial, and health-related factors influence walking behavior in disabled older women. DESIGN: Cross-sectional analyses of baseline data from the Women's Health and Aging Study (WHAS). SETTING: An urban community encompassing 12 contiguous zip code areas in the eastern portion of Baltimore City and part of Baltimore County, Maryland. PARTICIPANTS: A total of 920 moderately to severely disabled community-resident women, aged 65 years and older, identified from an age-stratified random sample of Medicare beneficiaries. MEASUREMENTS: Walking behavior was defined as minutes walked for exercise and total blocks walked per week. Independent variables included self-reported walking difficulty, sociodemographic factors, psychological status (depression, mastery, anxiety, and cognition), and health-related factors (falls and fear of falling, fatigue, vision and balance problems, weight, smoking, and cane use). RESULTS: Walking at least 8 blocks per week was strongly negatively related to severity of walking difficulty. Independent of difficulty level, older age, black race, fatigue, obesity, and cane use were also negatively associated with walking; living alone and high mastery had a positive association with walking. CONCLUSIONS: Even among functionally limited women, sociocultural, psychological, and health-related factors were independently associated with walking behavior. Thus, programs aimed at improving walking ability need to address these factors in addition to walking difficulties to maximize participation and compliance. PMID- 10366166 TI - Effects of shoe collar height and sole hardness on balance in older women. AB - OBJECTIVE: To determine whether shoe collar height and sole hardness affect balance in older women. DESIGN: A randomized order, cross-over, controlled comparison. SETTING: Intermediate care institution and regional hospital. PARTICIPANTS: Forty-two women aged 60 to 92 years (mean 76, SD = 9.03). The sample comprised 15 hostel (intermediate care) residents, 11 women who lived in a retirement village, and 16 women who lived independently in the community. OUTCOME MEASURES: Postural sway, maximal balance range and coordinated stability. MAIN RESULTS: The subjects underwent assessments of static balance (body sway) and dynamic balance (maximal balance range and coordinated stability) under five conditions: (1) in soft-soled bowls shoes, (2) in hard-soled bowls shoes, (3) in college-style shoes, (4) in college-style shoes with a high (boot) collar, and (5) barefoot. MANOVA analysis indicated that subjects were more stable when wearing the high collar shoes than when wearing the college shoes (P < .001) or when barefoot (P < .05). In contrast, subjects performed similarly in the balance tests in the soft and hard-soled shoes (P = .30) and no better than when barefoot (P = .12 and P = .93, respectively). CONCLUSIONS: The findings indicate that subjects had better balance when wearing shoes with high collars than when wearing shoes with low collars and that sole hardness was not related to balance. PMID- 10366167 TI - Endogenous levels of dehydroepiandrosterone sulfate, but not other sex hormones, are associated with depressed mood in older women: the Rancho Bernardo Study. AB - OBJECTIVE: The purpose of this study was to determine whether endogenous steroid hormone levels are associated with depressed mood in community-dwelling older women. DESIGN: A cross-sectional population-based study. SETTING: Rancho Bernardo, California PARTICIPANTS: A total of 699 non-estrogen using, community dwelling, postmenopausal women (aged 50 to 90 years) from the Rancho Bernardo cohort who were screened for depressed mood and had plasma obtained for steroid hormone assays in 1984-1987. MEASUREMENTS: Plasma levels of total and bioavailable (non-SHBG-bound) estradiol and testosterone, estrone, androstenedione, cortisol, dehydroepiandrosterone, and (DHEA) and its sulfate (DHEAS) were measured by radioimmunoassay. Mood and depression were assessed using the Beck Depression Inventory. RESULTS: Only DHEAS levels were significantly and inversely associated with depressed mood, and the association was independent of age, physical activity, and weight change (P = .0002). Age, sedentary lifestyle, and weight loss were positively associated with depressed mood. Alcohol intake, cigarette smoking, marital status, type of menopause, and season of testing were unassociated with depressed mood. A subset of 31 women with categorically defined depression had lower DHEAS levels compared with 93 age matched nondepressed women (1.17 +/- 1.08 vs 1.57 +/- .98 micromol/L; P = .01). CONCLUSIONS: These results add to the evidence that DHEA/S is a neuroactive steroid and point to the need for careful long-term clinical trials of DHEA therapy in older women with depressed mood. PMID- 10366168 TI - Geriatric syndromes as outcome measures of hospital care: can administrative data be used? AB - OBJECTIVE: To determine how often hospital administrative databases capture the occurrence of two common geriatric syndromes, pressure ulcers and incontinence. DESIGN: Retrospective comparison of a nursing home and hospital database. SETTING: Department of Veterans Affairs (VA) hospitals. PARTICIPANTS: All patients between 1992 and 1996 discharged from VA acute medical care and admitted to a VA nursing home. MEASUREMENTS: The presence of incontinence or a pressure ulcer (stage 2 or larger) on admission to the nursing home was determined. Hospital discharge diagnoses were then reviewed to determine whether these conditions were recorded. The effect of ulcer stage, total number of discharge diagnoses, and temporal trends on the recording of these conditions in discharge diagnoses was also noted. RESULTS: There were 17,004 admissions to nursing homes from acute care in 1996; 12.7% had a pressure ulcer and 43.4% were incontinent. Among these patients with a pressure ulcer, the hospital discharge diagnosis listed an ulcer in 30.8% of cases, and incontinence was included correctly as a discharge diagnosis in 3.4%. While deeper pressure ulcers were more likely to be recorded than superficial ulcers (P < .01), nearly 50% of stage 4 ulcers were not listed among hospital discharge diagnoses. Patients with more discharge diagnoses were more likely to record both conditions correctly. From 1992 to 1996, small but significant (P = .001) improvements were noted in the correct recording of these geriatric syndromes as discharge diagnoses. CONCLUSIONS: The occurrence of pressure ulcers and incontinence cannot be determined from hospital administrative databases and should not be used as outcomes when measuring quality of care among hospitalized patients. PMID- 10366169 TI - Home hospital program: a pilot study. AB - OBJECTIVE: To evaluate the basic safety and feasibility of hospital care at home (Home Hospital (HH)) for treating acutely ill older persons requiring hospitalization. DESIGN: Prospective case series SETTINGS AND PARTICIPANTS: Community-dwelling persons aged 65 and older requiring acute hospital admission for community-acquired pneumonia, chronic heart failure, chronic obstructive airways disease, or cellulitis. RESULTS: Seventeen subjects were treated in HH. One hundred twenty-two could not be enrolled because they presented for admission at times when HH was not operational. Six patients refused to enroll in HH. Subjects treated in HH had comparable clinical outcomes to those treated in the acute hospital and were highly satisfied with HH. Charges for HH care were 60% of those for the acute hospital care. CONCLUSIONS: In this pilot study, HH was safe, feasible, highly satisfactory, and cost-effective for certain acutely ill older persons who required acute hospitalization. PMID- 10366170 TI - Subclinical hypothyroidism in a biethnic, urban community. AB - OBJECTIVES: To evaluate the association between hypothyroidism, and the health status of older Hispanic and non-Hispanic white (NHW) men and women. To accomplish this, we determined the prevalences of the treated and untreated conditions and examined the associations between an elevated serum thyroid stimulating hormone (TSH) and cognitive and affective (mood) functions and the prevalences of symptoms and comorbidity, specifically coronary heart disease (CHD), diabetes, hypertension, and hyperlipidemia. DESIGN AND SETTING: A cross sectional study of equal numbers of Hispanic and NHW men and women selected randomly from the Health Care Financing Administration (Medicare) rolls and recruited for a home interview followed by a 4-hour interview/examination in a senior health clinic. PARTICIPANTS: 883 volunteers, mean age 74.1 years, participated in interviews/examinations MEASUREMENTS: Serum TSH was determined in 825 participants responding to questions about thyroid replacement therapy. Serum free thyroxine (free T4) concentrations were determined in 139 participants with elevated TSH concentrations (>4.6 microU/mL). Symptoms, cognitive tests, a screen for depression, comorbidities (e.g., CHD), and risk factors (e.g., lipid abnormalities, diabetes, and hypertension) were compared in participants with high versus normal TSH values. RESULTS: Subclinical hypothyroidism is more common in women than in men and in non-Hispanic white women compared with Hispanic women. No differences were observed between participants with TSH elevations from 4.7 to 10 microU/mL and those with normal TSH concentrations, and only a few differences were observed in those with TSH concentrations above 10. CONCLUSIONS: Subclinical hypothyroidism is a common condition in community-living older people, especially women. However, it appeared to have no effect on any of the measures of health status utilized until serum TSH concentrations exceeded 10 microU/mL, and even then the effects were rarely significant. PMID- 10366171 TI - Misreporting of total energy intake in older men and women. AB - Misreporting of total energy intake occurs frequently in dietary studies. Relatively few studies have been performed in older individuals who may be vulnerable to obesity and its associated health risks. In the present study, we examined misreporting of total energy intake by comparing self-reported food intake, measured by 3-day food diaries, to energy expenditure, measured with the doubly labeled water technique, in a relatively large sample of older men (n = 39) and women (n = 43). An additional objective was to identify potential predictors of misreporting, including body composition, fitness as assessed by peak VO2, and sociodemographic characteristics such as gender, living arrangement, education, and income. In general, men and women underreported total energy intake. The magnitude of the underreporting, as measured by percent difference between reported intake and measured total energy expenditure, was comparable between the sexes. Body mass index, waist circumference, and fat mass were significant correlates of underreporting of total energy intake, with heavier individuals underreporting more than leaner individuals. Among the demographic variables, living arrangement was a significant determinant of misreporting of total energy intake in older people. Individuals in marriage-like living arrangements underreported their total energy intake to a greater extent than married individuals. However, the magnitude of misreporting by those living alone did not differ from that of married individuals. The results of the present study highlight the need to examine misreporting of total energy intake in older individuals, who are more prone to obesity and its health risks. PMID- 10366172 TI - Lower incidence of tardive dyskinesia with risperidone compared with haloperidol in older patients. AB - OBJECTIVE: To compare the 9-month cumulative incidence of tardive dyskinesia (TD) with risperidone to that with haloperidol in older patients. DESIGN: A prospective longitudinal study. SETTING: An outpatient psychiatric clinic. PARTICIPANTS: Subjects were middle-aged and older (mean age 66 years) patients with schizophrenia, dementia, mood disorders, or other conditions with psychotic symptoms or severe behavioral disturbances. Sixty-one patients on risperidone were matched with 61 patients from a larger sample of haloperidol-treated patients in regard to age, diagnosis, and length of pre-enrollment neuroleptic intake to create clinically comparable groups. The median daily dose of each medication was 1.0 mg. MEASUREMENTS: Abnormal Involuntary Movement Scale, modified Simpson-Angus' scale for extrapyramidal symptoms, Brief Psychiatric Rating Scale, and Mini-Mental State Examination were administered at baseline, 1 month, and 3, 6, and 9 months. The diagnosis of TD was based on specific research criteria. The raters were blind to the patient's medication status. RESULTS: Life table analysis revealed that patients treated with haloperidol were significantly more likely to develop TD than patients treated with risperidone (P < .05, Peto Prentice). CONCLUSIONS: The atypical antipsychotic risperidone is significantly less likely to result in TD than the conventional neuroleptic haloperidol in a high-risk group of older patients, at least over a 9-month period. PMID- 10366173 TI - The efficacy of omeprazole-based short-term triple therapy in Helicobacter pylori positive older patients with dyspepsia. AB - OBJECTIVE: To evaluate the efficacy of 1-week triple therapy with omeprazole, clarithromycin,and tinidazole (OCT) in Helicobacter pylori-positive older patients with dyspepsia. DESIGN: A prospective, nonrandomized therapeutic study. SETTING: The primary care and referral center of a gastroenterological outpatient clinic at a central university hospital serving an urban population (>1 million) in Israel. PARTICIPANTS: The study group consisted of 134 patients (71 men, and 63 women) more than 60 years old who were referred for evaluation of symptoms of dyspepsia and were endoscopically diagnosed as H. pylori positive. The patients were divided into two groups: those who received their first course of anti-H. Pylori therapy during this study (Group 1) and those who had previously received standard metronidazole and bismuth combination therapies that failed to eradicate the H. pylori (Group 2). MEASUREMENTS: All the patients underwent upper gastrointestinal endoscopy, and H. pylori infection was confirmed by a rapid urease test (CUTest) and/or histological staining. Therapeutic efficacy was assessed by a 13C-urea breath test 4 weeks after completion of treatment. RESULTS: The mean age of the study population was 68.8 years (range 60-87). There were 112 patients in Group 1 and 22 patients in Group 2. Endoscopic findings were: gastritis (in 46), gastric ulcer (8), duodenal ulcer (52), and duodenitis (28). The H. pylori eradication rate was significantly higher in Group 1 patients (104/112, 92.9%) than in patients of Group 2 (15/22, 68.2%). There was no difference in the eradication rate in relation to gender, endoscopic diagnosis, more advanced age, place of birth, or smoking habits. The compliance in both groups was equally good, and no major side effects were recorded. CONCLUSIONS: A 1-week OCT triple therapy is well tolerated and effective as first line therapy for H. pylori among older people. It is less effective in patients previously treated. PMID- 10366174 TI - Low blood pressure and incidence of dementia in a very old sample: dependent on initial cognition. AB - OBJECTIVE: To examine whether initially low blood pressure is related to the incidence of dementia. DESIGN: A population-based prospective study. SETTING: The Kungsholmen district of Stockholm, Sweden PARTICIPANTS: Three hundred four nondemented subjects aged 75 to 96 years at baseline. MEASUREMENTS AND MAIN RESULTS: After an average of 3 years, 81 dementia cases were identified (67 with Alzheimer's disease cases). Compared with individuals with baseline systolic pressure of 141 to 179 mm Hg, those with systolic pressure < or = 140 mm Hg had a significantly higher risk of dementia (relative risk (RR) = 1.9, 95% confidence interval (CI), 1.2-3.2) and Alzheimer's disease (RR = 2.2, 95% CI, 1.2-3.8). However, the RR in relation to systolic pressure < or = 140 mm Hg was 1.3 (0.8 2.2) for all dementia and 1.5 (0.8-2.6) for Alzheimer's disease, when the baseline Mini-Mental State Examination (MMSE) score was included in the model as a dichotomous variable (< 24 vs > or = 24). Baseline MMSE < 24 significantly predicted the occurrence of dementia (RR = 6.9; 95% CI, 4.3-11.1). Systolic pressure < or = 140 mm Hg was significantly related to MMSE score < 24 at baseline. CONCLUSIONS: These data suggest that low blood pressure may be an early correlate of a dementing process although a causative effect cannot be definitely ruled out. PMID- 10366175 TI - Insulin effects on the left ventricle in older hypertensive subjects. AB - BACKGROUND: To evaluate the effects of hyperinsulinemia on left ventricular (LV) structure and function in older hypertensive subjects METHODS: Thirty-seven hypertensive subjects (17 men/20 women) aged 50 to 80, were studied. LV mass were evaluated echocardiographically according to the Penn convention. A 75-g oral glucose tolerance test (OGTT) was performed after overnight fasting, and both blood glucose and insulin concentrations were assayed at 0, 30, 60, 90, 120, and 180 minutes. Comparison between groups was performed by analysis of variance. A P value of .05 was considered statistically significant. RESULTS: When the hypertensive patients were divided into two groups according to the median value of 2-hour post-loading plasma insulin, there was no difference in blood pressure levels between the groups. However, hyperinsulinemic hypertensive subjects had an increased LV mass (P < .05), mean wall thickness, and interventricular septum thickness (P < .05 for both parameters) and had better systolic function-ejection and shortening fractions (P < .0001 for both indices). CONCLUSIONS: Hyperinsulinemia may be associated with increased left ventricular mass and with a better systolic performance in older hypertensive subjects. PMID- 10366176 TI - Definition of race and ethnicity in older people in Medicare and Medicaid. AB - BACKGROUND: Race and ethnicity are important predictors of health care access and outcomes, but quality of their documentation in the healthcare system is often problematic. OBJECTIVES: To study the agreement between Medicare and Medicaid descriptions of race and ethnicity in older beneficiaries. DESIGN: Quasiexperimental design in a natural practice setting. SETTING: New Jersey. PARTICIPANTS: 153,241 dually enrolled participants in Medicare and Medicaid. MEASUREMENTS: Agreement rates between administrative databases on recipients' race and ethnicity. RESULTS: Agreement between Medicare and Medicaid on the recipients' race and ethnicity was modest (kappa = .58; 95% CI, .57-.58) for men and women alike and across different age groups. Depending on whether Medicare or Medicaid was used as the reference standard, the relative agreement rates for race and ethnic group assignments varied. For example, using Medicare as the reference, the relative agreement rate was 84% for whites, 74% for blacks, 61% for others, 23% for Hispanics, and only 5% for Asians. Using Medicaid as the reference, a different pattern emerged. However, such gradients of agreement rates across racial groups were observed in both programs. Medicare and Medicaid reported different percentages of all race and ethnicity groups, with Medicaid reporting greater proportions of White and Black beneficiaries, and Medicare reporting greater proportions of Hispanic, Asian, and Other groups. CONCLUSIONS: Depiction of race and ethnicity data in large government health insurance programs is approximate at best and often contradictory from one program to another. This can impede efforts to study the relationship between these important characteristics and health care utilization and outcomes. PMID- 10366177 TI - Bilateral internuclear ophthalmoplegia: an initial presenting sign of giant cell arteritis. PMID- 10366178 TI - Management of the older person with atrial fibrillation. AB - OBJECTIVE: To review the management of the older person with atrial fibrillation (AF). DATA SOURCES: A computer-assisted search of the English language literature (MEDLINE) database followed by a manual search of the bibliographies of pertinent articles. STUDY SELECTION: Studies on the management of persons with AF were screened for review. Studies of persons older than age 60 and recent studies were emphasized. DATA EXTRACTION: Pertinent data were extracted from the reviewed articles. Emphasis was placed on studies involving older persons. Relevant articles were reviewed in depth. DATA SYNTHESIS: Available data about the management of persons with paroxysmal or chronic AF were summarized CONCLUSIONS: Management of AF includes treatment of the underlying disease and precipitating factors. Immediate direct-current cardioversion should be performed in persons with AF associated with an acute myocardial infarction, chest pain caused by myocardial ischemia, hypotension, severe heart failure, or syncope. Intravenous verapamil, diltiazem, or beta-blockers should be used to slow a very rapid ventricular rate associated with AF immediately. Oral verapamil, diltiazem, or a beta-blocker should be given if a rapid ventricular rate occurs at rest or during exercise despite digoxin. Amiodarone may be used in selected persons with symptomatic life-threatening AF refractory to other drug therapy. Nondrug therapies should be performed in persons with symptomatic AF in whom a rapid ventricular rate cannot be slowed by drug therapy. Paroxysmal AF associated with the tachycardia-bradycardia syndrome should be treated with a permanent pacemaker in combination with drugs. A permanent pacemaker should be implanted in persons with AF who develop cerebral symptoms such as dizziness or syncope associated with ventricular pauses greater than 3 seconds that are not drug-induced. Elective cardioversion of AF should not be performed in asymptomatic older persons with chronic AF. Unless transesophageal echocardiography has shown no thrombus in the left atrial appendage before cardioversion, oral warfarin should be given for 3 weeks before elective direct-current or drug cardioversion of AF and continued for at least 4 weeks after maintenance of sinus rhythm. Many cardiologists prefer the treatment strategy, especially in older persons, of ventricular rate control plus warfarin rather than maintaining sinus rhythm with antiarrhythmic drugs. Digoxin should be avoided in persons with sinus rhythm who have a history of paroxysmal AF. Older persons with chronic or paroxysmal AF who are at high risk for stroke or who have a history of hypertension and no contraindications to warfarin should receive long-term warfarin to achieve an International Normalized Ratio of 2.0 to 3.0. Older persons with AF who are at low risk for stroke or who have contraindications to warfarin should receive 325 mg of aspirin daily. PMID- 10366179 TI - Assessment and control for confounding by indication in observational studies. AB - In the evaluation of pharmacologic therapies, the controlled clinical trial is the preferred design. When clinical trial results are not available, the alternative designs are observational epidemiologic studies. A traditional concern about the validity of findings from epidemiologic studies is the possibility of bias from uncontrolled confounding. In studies of pharmacologic therapies, confounding by indication may arise when a drug treatment serves as a marker for a clinical characteristic or medical condition that triggers the use of the treatment and that, at the same time, increases the risk of the outcome under study. Confounding by indication is not conceptually different from confounding by other factors, and the approaches to detect and control for confounding--matching, stratification, restriction, and multivariate adjustment- are the same. Even after adjustment for known risk factors, residual confounding may occur because of measurement error or unmeasured or unknown risk factors. Although residual confounding is difficult to exclude in observational studies, there are limits to what this "unknown" confounding can explain. The degree of confounding depends on the prevalence of the putative confounding factor, the level of its association with the disease, and the level of its association with the exposure. For example, a confounding factor with a prevalence of 20% would have to increase the relative odds of both outcome and exposure by factors of 4 to 5 before the relative risk of 1.57 would be reduced to 1.00. Observational studies have provided important scientific evidence about the risks associated with several risk factors, including drug therapies, and they are often the only option for assessing safety. Understanding the methods to detect and control for confounding makes it possible to assess the plausibility of claims that confounding is an alternative explanation for the findings of particular studies. PMID- 10366180 TI - Serum levels of IL-6 and development of disability in older persons. PMID- 10366181 TI - Minor and subsyndromal depression: functional disability worth treating. PMID- 10366182 TI - Interesting conflicts and conflicting interests. PMID- 10366183 TI - Dementia assessment and CAT scans in primary care. PMID- 10366184 TI - Development of a clinical rating scale for persons with Parkinson's disease. PMID- 10366185 TI - Characteristics of AIDS in older patients: a follow-up study. PMID- 10366186 TI - Low complication rate after insertion of percutaneous endoscopic gastrostomy by a geriatrics-oriented team. PMID- 10366187 TI - Thoughts of a traveler. PMID- 10366188 TI - Excitotoxins in neuronal apoptosis and necrosis. AB - Neuronal loss is common to many neurodegenerative diseases. Although necrosis is a common histopathologic feature observed in neuropathologic conditions, evidence is increasing that apoptosis can significantly contribute to neuronal demise. The prevalence of either type of cell death, apoptosis or necrosis, and the relevance for the progression of disease is still unclear. The debate on the occurrence and prevalence of one or the other type of death in pathologic conditions such as stroke or neurotoxic injury may in part be resolved by the proposal that different types of cell death within a tissue reflect either partial or complete execution of a common death program. Apoptosis is an active process of cell destruction, characterized morphologically by cell shrinkage, chromatin aggregation with extensive genomic fragmentation, and nuclear pyknosis. In contrast, necrosis is characterized by cell swelling, linked to rapid energy loss, and generalized disruption of ionic and internal homeostasis. This swiftly leads to membrane lysis, release of intracellular constituents that evoke a local inflammatory reaction, edema, and injury to the surrounding tissue. During the past few years, our laboratories have studied the signals and mechanisms responsible for induction or prevention of apoptosis/necrosis in neuronal injury and this is the subject of this review. PMID- 10366189 TI - Noninvasive assessment of changes in cytochrome-c oxidase oxidation in human subjects during visual stimulation. AB - In this study the authors used a whole-spectrum near-infrared spectroscopy approach to noninvasively assess changes in hemoglobin oxygenation and cytochrome c oxidase redox state (Cyt-Ox) in the occipital cortex during visual stimulation. The system uses a white light source (halogen lamp). The light reflected from the subject's head is spectrally resolved by a spectrograph and dispersed on a cooled charge-coupled device camera. The authors showed the following using this approach: (1) Changes in cerebral hemoglobin oxygenation (increase in concentration of oxygenated hemoglobin, decrease in concentration of deoxygenated hemoglobin) in the human occipital cortex during visual stimulation can be assessed quantitatively. (2) The spectral changes during functional activation cannot be completely explained by changes in hemoglobin oxygenation solely; Cyt Ox has to be included in the analysis. Only if Cyt-Ox is considered can the spectral changes in response to increased brain activity be explained. (3) Cytochrome-c oxidase in the occipital cortex of human subjects is transiently oxidized during visual stimulation. This allows us to measure vascular and intracellular energy status simultaneously. PMID- 10366190 TI - Differential expressions of glycine transporter 1 and three glutamate transporter mRNA in the hippocampus of gerbils with transient forebrain ischemia. AB - The extracellular concentrations of glutamate and its co-agonist for the N-methyl d-aspartate (NMDA) receptor, glycine, may be under the control of amino acid transporters in the ischemic brain. However, there is little information on changes in glycine and glutamate transporters in the hippocampal CA1 field of gerbils with transient forebrain ischemia. This study investigated the spatial and temporal expressions of glycine transporter 1 (GLYT1) and three glutamate transporter (excitatory amino acid carrier 1, EAAC1; glutamate/aspartate transporter, GLAST; glutamate transporter 1, GLT1) mRNA in the gerbil hippocampus after 3 minutes of ischemia. The GLYT1 mRNA was transiently upregulated by the second day after ischemia in astrocytelike cells in close vicinity to hippocampal CA1 pyramidal neurons, possibly to reduce glycine concentration in the local extracellular spaces. The EAAC1 mRNA was abundantly expressed in almost all pyramidal neurons and dentate granule cells in the control gerbil hippocampus, whereas the expression level in CA1 pyramidal neurons started to decrease by the fourth day after ischemia in synchrony with degeneration of the CA1 neurons. The GLAST and GLT1 mRNA were rather intensely expressed in the dentate gyrus and CA3 field of the control hippocampus, respectively, but they were weakly expressed in the CA1 field before and after ischemia. As GLAST and GLT1 play a major role in the control of extracellular glutamate concentration, the paucity of these transporters in the CA1 field may account for the vulnerability of CA1 neurons to ischemia, provided that the functional GLAST and GLT1 proteins are also less in the CA1 field than in the CA3 field. This study suggests that the amino acid transporters play pivotal roles in the process of delayed neuronal death in the hippocampal CA1 field. PMID- 10366191 TI - Cerebrovascular hemodynamics and ischemic tolerance: lipopolysaccharide-induced resistance to focal cerebral ischemia is not due to changes in severity of the initial ischemic insult, but is associated with preservation of microvascular perfusion. AB - Lipopolysaccharide (LPS), administered 72 hours before middle cerebral artery (MCA) occlusion, confers significant protection against ischemic injury. For example, in the present study, LPS (0.9 mg/kg intravenously) induced a 31% reduction in infarct volume (compared with saline control) assessed 24 hours after permanent MCA occlusion. To determine whether LPS induces true tolerance to ischemia, or merely attenuates initial ischemic severity by augmenting collateral blood flow, local CBF was measured autoradiographically 15 minutes after MCA occlusion. Local CBF did not differ significantly between LPS- and saline pretreated rats (e.g., 34 +/- 10 and 29 +/- 15 mL x 100 g(-1) x min(-1) for saline and LPS pretreatment in a representative region of ischemic cortex), indicating that the neuroprotective action of LPS is not attributable to an immediate reduction in the degree of ischemia induced by MCA occlusion, and that LPS does indeed induce a state of ischemic tolerance. In contrast to the similarity of the initial ischemic insult between tolerant (LPS-pretreated) and nontolerant (saline-pretreated) rats, microvascular perfusion assessed either 4 hours or 24 hours after MCA occlusion was preserved at significantly higher levels in the LPS-pretreated rats than in controls. Furthermore, the regions of preserved perfusion in tolerant animals were associated with regions of tissue sparing. These results suggest that LPS-induced tolerance to focal ischemia is at least partly dependent on the active maintenance of microvascular patency and hence the prevention of secondary ischemic injury. PMID- 10366192 TI - Matrix metalloproteinases increase very early during experimental focal cerebral ischemia. AB - Microvascular integrity is lost during focal cerebral ischemia. The degradation of the basal lamina and extracellular matrix are, in part, responsible for the loss of vascular integrity. Matrix metalloproteinases (MMPs) may play a primary role in basal lamina degradation. By using a sensitive modification of gelatin zymography, the authors investigated the activity of MMP-2 and MMP-9 in frozen 10 microm sections of ischemic and nonischemic basal ganglia and plasma samples of 27 non-human primates after middle cerebral artery occlusion/reperfusion (MCAO/R) for various periods. The gelatinolytic activities were compared with parallel cell dUTP incorporation in the ischemic zones of adjacent sections. In the brain, the integrated density of MMP-2 increased significantly by 1 hour after MCAO and was persistently elevated thereafter. Matrix metalloproteinase-2 expression was highly correlated with the extent of neuron injury and the number of injured neurons (r = 0.9763, SE = 0.004, 2P < 0.0008). Matrix metalloproteinase-9 expression only was significantly increased in subjects with hemorrhagic transformation. In plasma, only MMP-9 increased transiently at 2 hours of MCAO. These findings highlight the early potential role of MMP-2 in the degradation of basal lamina leading to neuronal injury, and an association of MMP-9 with hemorrhagic transformation after focal cerebral ischemia. PMID- 10366193 TI - Inhibition of interleukin-1beta converting enzyme family proteases (caspases) reduces cold injury-induced brain trauma and DNA fragmentation in mice. AB - The authors examined the effect of z-VAD.FMK, an inhibitor that blocks caspase family proteases, on cold injury-induced brain trauma, in which apoptosis as well as necrosis is assumed to play a role. A vehicle alone or with z-VAD.FMK was administered into the cerebral ventricles of mice 15 minutes before and 24 and 48 hours after cold injury. At 24 hours after cold injury, infarction volumes in the z-VAD.FMK-treated animals were significantly smaller than infarction volumes in the vehicle-treated animals, and were further decreased at 72 hours (0.92 +/- 1.80 mm3, z-VAD.FMK-treated animals; 7.46 +/- 3.53 mm3, vehicle-treated animals; mean +/- SD, n = 7 to 8). The amount of DNA fragmentation was significantly decreased in the z-VAD.FMK-treated animals compared with the vehicle-treated animals, as shown by terminal deoxynucleotidyl transferase-mediated uridine 5' triphosphate-biotin nick end labeling staining and DNA gel electrophoresis. By Western blot analysis, both the proform and activated form of interleukin-1beta converting enzyme (caspase 1) were detected in the control brain, and the activated form showed moderate reduction after cold injury-induced brain trauma. These results indicate that caspase inhibitors could reduce cold injury-induced brain trauma by preventing neuronal cell death by DNA damage. The caspase family proteases appear to contribute to the mechanisms of cell death in cold injury induced brain trauma and to provide therapeutic targets for traumatic brain injury. PMID- 10366194 TI - A potential role for erythropoietin in focal permanent cerebral ischemia in mice. AB - The present study describes, for the first time, a temporal and spatial cellular expression of erythropoietin (Epo) and Epo receptor (Epo-R) with the evolution of a cerebral infarct after focal permanent ischemia in mice. In addition to a basal expression of Epo in neurons and astrocytes, a postischemic Epo expression has been localized specifically to endothelial cells (1 day), microglia/macrophage like cells (3 days), and reactive astrocytes (7 days after occlusion). Under these conditions, the Epo-R expression always precedes that of Epo for each cell type. These results support the hypothesis that there is a continuous formation of Epo, with its corresponding receptor, during the active evolution of a focal cerebral infarct and that the Epo/Epo-R system might be implicated in the processes of neuroprotection and restructuring (such as angiogenesis and gliosis) after ischemia. To support this hypothesis, a significant reduction in infarct volume (47%; P < 0.0002) was found in mice treated with recombinant Epo 24 hours before induction of cerebral ischemia. Based on the above, we propose that the Epo/Epo-R system is an endogenous mechanism that protects the brain against damages consequent to a reduction in blood flow, a mechanism that can be amplified by the intracerebroventricular application of exogenous recombinant Epo. PMID- 10366195 TI - Neurofilament proteolysis after focal ischemia; when do cells die after experimental stroke? AB - To determine the occurrence and time-course of presumably irreversible subcellular damage after moderate focal ischemia, rats were subjected to 1, 3, 6, 9, or 24 hours of permanent unilateral middle cerebral and common carotid occlusion or 3 hours of reversible occlusion followed by 3, 6, or 21 hours of reperfusion. The topography and the extent of damage were analyzed with tetrazolium staining and immunoblot using an antibody capable of detecting breakdown of neurofilament. Neurofilament proteolysis began after 3 hours in the infarct core but was still incomplete in penumbral regions up to 9 hours. Similarly, tetrazolium-staining abnormalities were observed in the core of 50% of animals after 3 hours of ischemia. At 6 hours of permanent ischemia, infarct volume was maximal, and further prolongation of occlusion to 9 or 24 hours did not increase abnormal tetrazolium staining. In contrast to permanent ischemia and in agreement with the authors' previous demonstration of "reperfusion injury" in this model, prolongation of reperfusion from 3 hours to 6 and 21 hours after 3 hours of reversible occlusion gradually augmented infarct volume by 203% and 324%, respectively. Neurofilament proteolysis initiated approximately 3 hours after ischemia was quantitatively greatest in the core and extended during reperfusion to incorporate penumbra with a similar time course to that of tetrazolium abnormalities. These data demonstrate that, at least as measured by neurofilament breakdown and mitochondrial failure, extensive cellular damage is not present in penumbral regions for up to 9 hours, suggesting the potential for rescuing these regions by appropriate and timely neuroprotective strategies. PMID- 10366196 TI - Age-dependent increase in ischemic brain injury in wild-type mice and in mice lacking the inducible nitric oxide synthase gene. AB - The authors investigated the influence of age on the outcome of cerebral ischemia in wild-type mice and in mice with a deletion of the inducible nitric oxide synthase (iNOS) gene. The middle cerebral artery was permanently occluded in iNOS null mice and in wild-type (C57BL/6) controls aged 4, 8, 16, and 24 weeks. Infarct volume was determined in thionin-stained brain sections 4 days after permanent middle cerebral artery occlusion. No differences in forebrain volume were found among wild-type and iNOS-null mice at the ages studied (P > 0.05). In C57BL/6 mice (n = 5 to 6/group), neocortical infarct volume corrected for swelling was 28 +/- 5 mm3 in 4-week-old mice, 28 +/- 3 at 8 weeks, 35 +/- 4 at 16 weeks, and 37 +/- 6 at 24 weeks (mean +/- SD). iNOS-null mice (n = 5 to 6/group) had smaller infarcts than wild-type controls at all ages (P < 0.05). However, the magnitude of the reduction was greater in 4-week-old (-29% +/- 10%) or 8-week-old mice (-24% +/- 8%), than in 16-week-old (-14% +/- 10%) or 24-week-old mice (-11% +/- 6%). Neurologic deficit scores improved significantly between 24 and 96 hours in 4- and 8-week-old iNOS-null mice compared with age-matched wild-type mice (P < 0.05). However, in 16- or 24-week-old iNOS-null mice, neurologic deficits did not improve (P > 0.05). The authors conclude that in iNOS-/- and in wild-type mice, the size of the infarct produced by occlusion of the middle cerebral artery is larger in older than in younger mice. However, the reduction in infarct volume observed in iNOS-null mice is age-dependent and is greatest at 1 to 2 months of age. Therefore, age is a critical variable in studies of focal cerebral ischemic damage, both in wild-type mice and in mouse mutants. PMID- 10366197 TI - Dynamics of nitrotyrosine formation and decay in rat brain during focal ischemia reperfusion. AB - The purpose of this study was to establish the dynamics of nitrotyrosine (NO2 Tyr) formation and decay during the rise of NO2-Tyr in rat brain subjected to 2 hour focal ischemia-reperfusion, and to evaluate the role of inducible nitric oxide synthase in the rise. The authors first determined the half life of NO2-Tyr in rat brain at 24 hours after the start of reperfusion by blocking NO2-Tyr formation with N(G)-monomethyl-L-arginine and after the decay of NO2-Tyr by means of a hydrolysis/HPLC procedure. The values obtained were approximately 2 hours in both peri-infarct and core-of-infarct regions. Using the same hydrolysis/HPLC procedure, the ratio of nitrotyrosine to tyrosine from the 2-hour occlusion to as much as 72 hours after the start of reperfusion was measured in the presence and absence of aminoguanidine (100 mg/kg intraperitoneally twice a day). In the absence of aminoguanidine, the ratio of NO2-Tyr in the peri-infarct and core-of infarct regions reached 0.95% +/- 0.34% and 0.52% +/- 0.34%, respectively, at 1 hour after the start of reperfusion. The elevated levels persisted until 48 hours, then declined. The peri-infarct region showed the highest percent NO2-Tyr level, followed by the core of infarct, then the caudoputamen. Aminoguanidine significantly reduced NO2-Tyr formation (up to 90% inhibition) during 24 to 48 hours. The authors conclude that inducible nitric oxide synthase is predominantly responsible for NO2-Tyr formation, at least in the late phase of reperfusion. These results have important implications for the therapeutic time window and choice of nitric oxide synthase inhibitors in patients with cerebral infarction. PMID- 10366198 TI - The effect of the nitric oxide synthase inhibitor L-NMMA on basal CBF and vasoneuronal coupling in man: a PET study. AB - Nitric oxide (NO) regulates basal CBF. In a number of animal models NO has been implicated in the mediation of the regional changes in CBF (rCBF) that accompany neuronal activation (vasoneuronal coupling). However, some results in animal models have failed to confirm this finding, and the validity of extrapolation to man from animal data is uncertain. To determine the contribution of NO to basal global CBF and activation-induced changes in rCBF, the authors have performed quantitative H2(15)O positron emission tomography (PET) studies before and after administration of the non-isoform-specific NO synthase inhibitor, N(G)-monomethyl L-arginine (L-NMMA), in 10 healthy male volunteers. Learning a novel sequence of finger movements was used as a paradigm to induce regional frontal cortex activation. The effect of NO synthase inhibition on the magnitude and pattern of activation was determined. Resting global CBF fell from 33.3 +/- 5.3 mL x 100 g( 1) x min(-1) at rest before L-NMMA, to 26.5 +/- 7.7 mL x 100 g(-1) x min(-1) after L-NMMA (P = 0.001). This fall was reversed by L-arginine administration. Learning sequential finger movements induced increases in rCBF in the left motor, right prefrontal, and bilateral premotor cortices. After NO synthase inhibition with L-NMMA, there was no change in this pattern of activation and no reduction in the magnitude of rCBF responses at the foci of maximal stimulation before and after L-NMMA. These findings confirm that NO production contributes to basal CBF regulation in man, but show that systemic NO synthase inhibition with L-NMMA does not impair regional vasoneuronal coupling. PMID- 10366199 TI - Evidence of a cerebrovascular postarteriole windkessel with delayed compliance. AB - A pronounced temporal mismatch was observed between the responses of relative cerebral blood volume (rCBV) measured by magnetic resonance imaging and relative cerebral blood flow measured by laser-Doppler flowmetry in rat somatosensory cortex after electrical forepaw stimulation. The increase of relative cerebral blood flow after stimulus onset and decrease after stimulus cessation were accurately described with a single exponential time constant of 2.4 +/- 0.8 seconds. In contrast, rCBV exhibited two distinct and nearly sequential processes after both onset and cessation of stimulation. A rapid change of rCBV (1.5 +/- 0.8 seconds) occurring immediately after onset and cessation was not statistically different from the time constant for relative cerebral blood flow. However, a slow phase of increase (onset) and decrease (cessation) with an exponential time constant of 14 +/- 13 seconds began approximately 8 seconds after the rapid phase of CBV change. A modified windkessel model was developed to describe the temporal evolution of rCBV as a rapid elastic response of capillaries and veins followed by slow venous relaxation of stress. Venous delayed compliance was suggested as the mechanism for the poststimulus undershoot in blood oxygen-sensitive magnetic resonance imaging signal that has been observed in this animal model and in human data. PMID- 10366200 TI - Modeling cerebral blood flow and flow heterogeneity from magnetic resonance residue data. AB - Existing model-free approaches to determine cerebral blood flow by external residue detection show a marked dependence of flow estimates on tracer arrival delays and dispersion. In theory, this dependence can be circumvented by applying a specific model of vascular transport and tissue flow heterogeneity. The authors present a method to determine flow heterogeneity by magnetic resonance residue detection of a plasma marker. Probability density functions of relative flows measured in six healthy volunteers were similar among tissue types and volunteers, and were in qualitative agreement with literature measurements of capillary red blood cell and plasma velocities. Combining the measured flow distribution with a model of vascular transport yielded excellent model fits to experimental residue data. Fitted gray-to-white flow-rate ratios were in good agreement with PET literature values, as well as a model-free singular value decomposition (SVD) method in the same subjects. The vascular model was found somewhat sensitive to data noise, but showed far less dependence on vascular delay and dispersion than the model-free SVD approach. PMID- 10366201 TI - Sentinel lymph node biopsy for breast cancer: the role of previous biopsy on patient eligibility. AB - Several reports have demonstrated the accurate prediction of axillary nodal status with radiolocalization and selective resection of sentinel lymph nodes (SLNs) in patients with breast cancer (BC). Because of concerns over lymphatic disruption, several authors have proposed that prior excisional breast biopsy is a contraindication for SLN biopsy. Clear unfiltered 99mtechnetium-sulfur colloid (1.0 mCi) was injected around the perimeter of the breast lesion (palpable and nonpalpable) or prior biopsy site. Resection of the radiolocalized SLN was then performed. Axillary lymph node dissection was performed immediately after SLN biopsy in the first 57 patients. Eighty-two BC patients underwent SLN biopsy. The SLN was localized in 98 per cent (80 of 82). The type of previously performed diagnostic biopsy or the location of the primary lesion did not influence the ability to localize the sentinel lymph node. In the 57 patients who had axillary lymph node dissection, metastatic disease was identified in 23 per cent (13 of 57). Axillary nodal status was accurately predicted in 98 per cent (56 of 57). Early experience with radiolocalization and selective resection of SLN in BC remains promising. By demonstrating the effective localization of the SLN regardless of the extent of prior biopsy, these data support expanding the number of patients potentially eligible for SLN biopsy. PMID- 10366202 TI - Sentinel node biopsy and cytokeratin staining for the accurate staging of 478 breast cancer patients. AB - Sentinel lymph node (SLN) mapping is an effective and accurate method of sampling the axillary nodal basin for metastatic disease. The SLN is the first node to receive afferent lymphatic drainage from the primary tumor. Lymphatic mapping and SLN biopsy have allowed pathologists to perform a more detailed examination of the SLN(s) and, therefore, provide more accurate staging of the regional lymphatic basin. Recently, more sensitive assays have been developed to increase the detection rate of micrometastatic to the axillary lymph nodes. Cytokeratin (CK) immunohistochemical (IHC) staining of the SLN detects micrometastatic disease, which is frequently missed on routine hematoxylin and eosin (H&E) histology. Therefore, lymphatic mapping combined with CK IHC staining of the SLN provides more accurate staging of the regional lymph nodes in patients with breast cancer. At Moffitt Cancer Center, 478 patients with newly diagnosed breast cancer underwent intraoperative lymphatic mapping using a combination of vital blue dye and technetium-labeled sulfur colloid. The excised SLNs were examined grossly, by intraoperative imprint cytology, by standard H&E histology, and by IHC stains for CK. SLNs that were only CK positive were confirmed malignant by sectioning the block, staining with H&E and finding cells with malignant cytology. Lymphatic mapping and CK IHC staining of the SLNs was successfully performed in 478 newly diagnosed breast cancer patients. Twenty-eight patients had unsuccessful lymphatic mapping for an overall failure rate of 5.5 per cent. A total of 134 (28%) patients had positive nodes (N1) detected. Ninety-three of these patients had both H&E and CK-positive lymph nodes, and an additional 41 patients had only CK-positive SLN(s). A total of 385 patients had H&E-negative SLNs, but only 344 patients had negative SLN(s) defined as both H&E and CK negative. Therefore, 41 (10.6%) of the 385 H&E-negative patients were upstaged, because of the detection of malignant cells by cytokeratin IHC staining of the SLN. Microstaging of SLNs with CK has shifted 10.6 per cent of our patient population from stage I to stage II disease. Undetected micrometastatic disease to the regional lymph nodes may account for the significant proportion of stage I breast cancer treatment failures. Furthermore, the ability to accurately stage the axilla by using lymphatic mapping techniques, SLN biopsy, and more sensitive assays may help identify a subgroup of truly node-negative patients with invasive breast cancer who can avoid the morbidity associated with a complete axillary dissection or systemic chemotherapy. Finally, those patients found to have micrometastatic disease to the regional lymph nodes can be treated appropriately in a more selective fashion. PMID- 10366203 TI - Clinical characteristics and antibiotic utilization in surgical patients with Clostridium difficile-associated diarrhea. AB - Clostridium difficile-associated diarrhea (CDAD) remains a significant problem in surgical patients. To address this, we prospectively studied all episodes of treated CDAD in surgical inpatients at the University of Virginia hospital from December 1996 through March 1998. CDAD accounted for 3.2 per cent (32) of 1000 total infections. Compared with a randomly selected control group with other nosocomial infections, patients with CDAD had a longer period from the time of admission to diagnosis of infection (19 +/- 4 versus 9 +/- 1; P = 0.01), were more likely to be female (66% versus 37%; P = 0.009), and had a higher overall crude mortality (31% versus 11%; P = 0.01), although there were no deaths directly attributable to CDAD. Ciprofloxacin (19%) and cefoxitin (16%) were the most common individual antibiotics prescribed before the diagnosis of CDAD. The average time from completion of antibiotic therapy to diagnosis of CDAD was 7 +/- 2 days (range, 0-58). Sixteen per cent (5 of 32) developed CDAD after administration of prophylactic perioperative antibiotics only. The high crude mortality rate associated with CDAD suggests that this may be a significant predictor of poor outcome among infected surgical patients. Antibiotics used commonly but not classically associated with CDAD frequently precipitate this infection. Finally, the use of prophylactic antibiotics is not without risk, as demonstrated by the significant percentage of CDAD occurring after routine administration of these agents. PMID- 10366204 TI - The pitfalls of establishing a statewide vascular registry: the South Carolina experience. AB - Concerned about the inadequacy of a centralized database and the importance of low morbidity and mortality on carotid endarterectomy efficacy, the South Carolina Vascular Surgical Society prospectively instituted a computer registry for carotid procedures performed by its members, to establish a statewide standard of practice. From January 1994 through December 1997, 23 of the 30 physician members voluntarily registered data on 1652 carotid operations at 14 hospitals into a central database. Blinded results were reviewed biannually. Complete data (1995-1997) were available for 1199 cases. The patients tended to be >64 years old (72%), male (62%), and white (93%). Carotid endarterectomy was the most frequently performed operation (90%). Perioperative complications (< or = 30 days) occurred in 173 patients (14.4%), including stroke (n = 19; 1.6%), death (n = 8; 0.7%), and stroke/death (n = 25; 2.0%). Although 23 surgeons (77% of the society) contributed some data, only 10 surgeons (33%) contributed complete data on >10 patients/year. Despite biannual efforts to boost participation, case entry remained stable (1994, 358; 1995, 347; 1996, 425; and 1997, 427), representing about one-third of the estimated carotid procedures performed in the state during that period. The cost of the registry was approximately $11,500. Audit of 8 surgeons revealed a >95 per cent match against the statewide discharge database and low error rate versus independent medical record review. This experience confirms that excellent outcomes after carotid endarterectomy are not limited to a few select centers and can be accomplished by adequately trained surgeons in a variety of institutional settings. Incomplete physician participation, however, inevitably raises questions about the utility of such efforts. Until volunteer registries induce full participation by heightening perceived physician benefit, their role will remain limited for future outcomes research. PMID- 10366205 TI - "Blind" placement of long-term central venous access devices: report of 589 consecutive procedures. AB - Placement of long-term central venous access devices, such as Hickman catheters and implanted subcutaneous ports, has traditionally been performed in the operating room with fluoroscopy. This study reports our experience with percutaneous placement of these devices in the outpatient clinic setting without the use of real-time imaging. Results were generated from a prospective database of all adult patients undergoing placement of central venous access in the outpatient clinic of the Wake Forest University Baptist Medical Center. This database revealed that during the years 1996 and 1997, long-term central venous catheter placement was attempted in 589 adult patients in the outpatient clinic. Technical success was achieved in 558 patients (92%). This included 278 tunneled catheters and 280 totally implanted devices. Repositioning of the catheter tip was required in 16 patients (2.9%). The incidence of pneumothorax was 1.9 per cent. Late complications, including infection and thrombosis, occurred in 9 per cent. The average procedure-related charge for placement of a single-lumen central venous port in the outpatient clinic was $1691 versus $4559 in the operating room and $3890 in the radiology department. We conclude that routine placement of long-term central venous access devices in the outpatient clinic, without the use of real-time imaging, yields acceptable success rates and may have economic advantages over procedures performed in the operating room or radiology department. PMID- 10366206 TI - Outpatient laparoscopic cholecystectomy: what predicts the need for admission? AB - Laparoscopic cholecystectomy (LC) is commonly performed as an outpatient (OP) procedure in selected patients, either in ambulatory surgery units associated with a hospital or in freestanding facilities. To identify factors that may preclude OPLC, a retrospective analysis of all patients who underwent LC by two surgeons from August 1996 through June 1998 was performed. A total of 126 patients were divided into three groups. Group I comprised 102 patients who underwent attempted elective OPLC. Group II comprised 20 patients who underwent LC on an emergent basis. Group III comprised 5 patients who were admitted before LC. Data were gathered regarding patient demographics, preoperative diagnoses, preoperative laboratory values, length of stay after surgery, and complications. These data were analyzed using logistic regression and univariate analysis. Age >60 and American Society of Anesthesiologists (ASA) class >2 appeared to be significant predictors of admission, but when considered together, neither was significant. The diagnosis of acute cholecystitis or biliary pancreatitis was highly predictive of admission in both groups. An ASA class >2 did predict postoperative stay of more than 12 hours. These data suggest that OPLC can be performed safely in unselected patients. However, those patients with an ASA class >2 or with a diagnosis of biliary pancreatitis or acute cholecystitis are more likely to require admission or postoperative stay over 12 hours, and these criteria should be considered relative contraindications to OPLC in free-standing facilities. PMID- 10366207 TI - Management of bile duct stones in 1572 patients undergoing laparoscopic cholecystectomy. AB - Evidence of bile duct stones (BDSs) was identified on routine cholangiogram in 136 (8.7%) of 1572 patients undergoing laparoscopic cholecystectomy from March 1989 through March 1997. Forty-two (30.9%) were unsuspected. All patients with evidence of BDSs underwent laparoscopic bile duct exploration (LBDE). Initially, a standard choledochotomy with T-tube drainage as in the open approach was used. Later, transcystic duct exploration was added to the algorithm. The algorithm evolved into an ongoing treatment protocol study that was initiated in March 1992. Through March 1997, 100 patients underwent LBDE based on the protocol. The study is divided into two groups. Group A comprises the total 136 patients undergoing LBDE, including those in the protocol study. A subgroup, Group B, comprises only the 100 patients in the protocol study. In Group A, LBDE was successful in 114 patients (83.8%). Stones were missed in seven patients and left behind for spontaneous passage or later retrieval in six patients. Eleven patients (8.1%) were converted to open. There were 13 major complications (9.6%), including the seven missed stones and two deaths. In Group B, LBDE was successful in 94 per cent. Stones were missed in one patient and intentionally left behind in four patients. One patient was converted to open. There were seven major complications (7%), including one of the missed stones and one death. Using the protocol algorithm and the techniques described, BDSs can be effectively managed laparoscopically at the time of cholecystectomy in approximately 94 per cent of cases. PMID- 10366208 TI - Post-treatment dopexamine infusions partially reverse reductions in cranial mesenteric blood flow and mucosal oxygenation induced by hypoxia in newborn piglets. AB - A severe hypoxic insult is known to induce dramatic reductions in newborn intestinal blood flow and is, thus, considered a vector for the development of neonatal intestinal ischemic diseases. Dopexamine (DPX) is a novel synthetic agent that has potent B2-adrenoceptor and dopaminergic activity, the clinical effects of which include an increase in cardiac output and in mesenteric blood flow. Having previously shown that infusion of DPX before hypoxia (HYP) mitigated the reduction in newborn mesenteric blood flow, we sought to define its efficacy when given after an established hypoxic insult. Ultrasonic transit time blood flow probes were placed around the ascending aorta and cranial mesenteric artery of anesthetized, mechanically ventilated 0 to 2-day-old piglets. Small bowel mucosal oxygenation was observed with a tissue oxygen monitoring system. After stabilization, animals were subjected to one of the following: HYP (FIO2 = 0.12) for 60 minutes (n = 12); DPX (5 microg/kg/min) infusion begun 10 minutes after induction of HYP/DPX (n = 11). Almost no alterations in any of the monitored variables were shown in a group (n = 5) of similarly instrumented, untreated animals. In contrast, although both hypoxic piglet groups experienced significant (P < 0.05, analysis of variance) declines from baseline cardiac output, mesenteric blood flow, and mucosal oxygenation, each of these deleterious effects was significantly (P < 0.05) blunted in the DPX-treated animals. During periods of systemic hypoxemia, the reductions in neonatal mesenteric blood flow and oxygenation can be somewhat blunted by DPX. As such, this agent may prove of clinical benefit when an infant is threatened by a hypoxic episode. PMID- 10366209 TI - Ultrasound: impact on diagnostic peritoneal lavage, abdominal computed tomography, and resident training. AB - Our objective was to determine the impact of abdominal ultrasound (US) on 1) the use of diagnostic peritoneal lavage (DPL) and abdominal computed tomography (ACT) for diagnosing blunt abdominal trauma (BAT) and on 2) surgical resident training. The study design was a retrospective chart review. Patients sustaining BAT who had ACT or DPL done during the 1-year period before the introduction of US (pre US) were compared with those from a 1-year period beginning 6 months after US (post-US). Data collected included diagnostic modality, demographic data, mortality, associated injuries, length of stay, mechanism of injury, and number of exploratory laparotomies. Of 128 patients in the pre-US group, 35 patients (27%; P < 0.001) underwent DPL, 0 patients (0%; P < 0.001) received US, and 92 patients (72%) received ACT, with positive results for 31 patients (34%). Exploratory laparotomy was performed on 35 patients (27%) in the pre-US group. Of 140 patients in the post-US group, 8 patients (6%; P < 0.001) underwent DPL, 120 patients (85%; P < 0.001) received US, and 108 patients (77%) received ACT, with positive results for 44 patients (42%). Exploratory laparotomy was performed on 22 patients (15%; P < 0.001) in the post-US group. Resident experience with DPL before and after the introduction of US and availability of US for graduated residents was documented. Chi-square and Fisher's exact test were used for statistical analysis. Resident experience changed from 22 to 3 DPLs per year in the pre- and post-US groups, respectively. Ten per cent of graduating residents had US available for use after leaving this institution. US replaced DPL and resulted in slightly more positive ACT scans in assessing BAT at our institution. Paradoxically, only 10 per cent of graduating residents had US available after leaving this institution. Until the use of US for diagnosing BAT has widespread use in the community, we must question our adequacy of resident preparation for diagnosing BAT. PMID- 10366210 TI - Surgical experience with hepatic colorectal metastasis. AB - The outcome of 134 patients undergoing hepatic resection for colorectal metastasis was studied. Current follow-up was available in 98 per cent of patients, for more than 5 years in 58 patients, and totaling 360 patient-years. Patients (52% male) had an average age of 62 +/- 1 years (standard error of the mean). Time lapse between the primary colon surgery and hepatic resection was a median of 16 months and a mean of 19 +/- 1 months. Thirty-two (24%) were operated on within 6 months for both their primary tumor and hepatic metastasis. Intensive care unit and total hospital length of stay were a median of 1 and 7 days, respectively. Pathology reports demonstrated that on average there were 2.0 +/- 0.1 lesions, with the largest lesion measuring 4.4 +/- 0.2 cm. In 72 per cent of patients, the lesions were found in one lobe only. CEA was elevated in 83 per cent of patients preoperatively and was 60 +/- 11 ng/mL before and 4.0 +/- 0.5 ng/mL after hepatic resection. Patient survival was 81 per cent at 1 year, 50 per cent at 3 years, 36 per cent at 5 years, and 23 per cent at 10 years. Actual 5- and 10-year survival was 22 of 58 (38%) patients and 4 of 21 (19%) patients respectively. Disease-free survival was 58 per cent at 1 year, 27 per cent at 3 years, 16 per cent at 5 years, and 12 per cent at 7 years. Survival was much better for one to four lesions than for five or more lesions (P < 0.01). Several other potential risk factors did not affect survival, including whether the patient received chemotherapy after hepatic resection. There were 36 (43%) patients who recurred with hepatic involvement only, 27 (32%) including hepatic involvement and 21 (25%) with nonhepatic involvement only. There were 15 patients who went on to receive repeat hepatic resections, with a 5-year survival of 74 per cent and disease-free survival of 58 per cent. Hepatic resection provides the best outcome of any form of therapy for selected patients with isolated hepatic metastasis. PMID- 10366211 TI - Combined blunt cardiac and pericardial rupture: review of the literature and report of a new diagnostic algorithm. AB - The spectrum of blunt cardiac injury varies from the asymptomatic cardiac concussion to the immediately fatal cardiac rupture. Although the majority of victims sustaining blunt cardiac rupture die before receiving medical attention, some survive to evaluation. The diagnosis of cardiac rupture, if established, typically results from the signs and symptoms of pericardial tamponade. However, some patients may have remarkably few signs and symptoms suggestive of cardiac injury and represent a significant diagnostic challenge. We provide two cases of cardiac rupture in which the diagnosis was delayed by the presence of an associated pericardial tear with decompression into the mediastinum and pleural space. In neither of the cases did existing institutional algorithms for blunt cardiac injury assist in establishing the diagnosis before the acute demise of the patient. The presence of a coexisting pericardial injury in these patients with blunt cardiac rupture obscured the diagnosis, leading to the deaths of these patients. A discussion of these two cases and review of the literature is provided with recommendations for diagnostic algorithms in patients sustaining blunt thoracic trauma with possible cardiac and pericardial injury. PMID- 10366212 TI - Organ injury scaling system can be used to predict length of stay in patients with penetrating neck injuries. AB - Predicting probability of survival of trauma patients has received greater attention than predicting other trauma outcomes such as length of stay. Most trauma scoring systems depend on the anatomic description of injuries of the Abbreviated Injury Score (AIS). The recently introduced Organ Injury Scale (OIS) was developed to give more precise and comprehensive anatomic description of injuries. Unlike the AIS, it also avoids including immediate injury sequelae, such as the amount of hemorrhage. This retrospective study was performed to assess the degree of association between the sum of grades of penetrating neck injuries using the OIS and the length of hospital stay. There were 31 males and 7 females, with ages ranging from 13 to 65 years and a mean of 31 years. The length of hospital stay ranged between 1 and 34 days, with a mean of 7.0 +/- 6.5 days. There were 32 vascular, 7 esophageal, and 7 tracheal injuries. The sum of the OIS grades correlates significantly with the length of hospital stay (r = 0.78; P < 0.005). This study suggests that the OIS system can be used to predict the length of hospital stay. It proposes also that the OIS replaces similar anatomic components of the AIS-based scoring systems. PMID- 10366213 TI - Giant colonic diverticulum: report of a case. AB - Giant colonic diverticulum is a rare complication of colonic diverticulosis. It typically occurs as a single diverticulum located on the antimesenteric border of the sigmoid colon. The most widely accepted theory for its development attributes the progressive dilation to a "ball-valve" mechanism, allowing air to enter but not to exit. Patients usually present complaining of abdominal pain and/or an abdominal mass, although they may remain asymptomatic. Physical examination reveals a tympanic abdominal mass that appears as a round radiolucency on plain radiographs and CT. Barium enema demonstrates the relationship of the diverticulum to bowel and may document communication with the colonic lumen. To alleviate symptoms and prevent complications, the recommended treatment is excision of the diverticulum in continuity with the involved colonic segment. We report a case and discuss the presentation, diagnosis, and management of giant colonic diverticulum. PMID- 10366214 TI - Antiperistaltic Roux-en-Y biliary-enteric bypass after bile duct injury: a technical error in reconstruction. AB - Bilioenteric reconstruction using a Roux limb of jejunum is a well-established surgical option for the reconstruction of the proximal bile duct. Previous studies discussing short- and long-term complications of biliary-enteric anastomosis have focused on technical aspects, such as the use of anastomotic stenting or the level of the biliary tree used. We report two cases of previously unreported complications after hepaticojejunostomy that resulted from a technical error in constructing the Roux limb. Within a 3-month period, two patients were referred to our institution with recurrent cholangitis after biliary reconstruction for injuries sustained during laparoscopic cholecystectomy. Reexploration disclosed major technical flaws in the construction of the Roux limb used for biliary drainage. Antiperistaltic limbs had been constructed in both patients: one from the distal ileum and one from the conventional location in the jejunum. In both cases, isoperistaltic reconstruction of the Roux limbs resolved the recurrent cholangitis. Cholangitis after biliary-enteric bypass can arise from a variety of etiologies and lead to anastomotic narrowing or ineffective drainage of the biliary tree. Review of the literature failed to disclose reports of technically flawed Roux limb construction as a cause of cholangitis. We present these cases to highlight the devastating consequences of antiperistaltic construction of the Roux limb. We hope that by publishing the role of this avoidable error in recurrent cholangitis after biliary-enteric bypass we may help prevent its future occurrence. PMID- 10366215 TI - Mature presacral teratoma in an adult male: a case report. AB - Presacral teratomas are rare tumors derived from more than one embryonic germ layer and are usually diagnosed in infancy. It has been estimated that the incidence of presacral teratoma in children ranges from 1 in 30,000 to 1 in 43,000 live births. However, the diagnosis of these tumors in adults is extremely rare. H. Head et al. reviewed the world literature in 1975 and found only 71 reported cases. Since that time, an additional 14 cases have been reported, and only one of these was found in the United States. This study reports a case of an adult male who presented with recurrent infected pilonidal cysts that proved to be benign presacral teratoma at biopsy. The patient underwent resection by left hemisacrectomy and primary closure using a posterior approach and, since this procedure, has had no evidence of recurrence. The case is presented along with a review of the relevant literature. PMID- 10366216 TI - Peripheral clear cell cholangiocarcinoma: a rare histologic variant. AB - We present the case of a 50-year-old diabetic male who underwent open cholecystectomy for acute gangrenous cholecystitis. At the time of exploration, a 1.5-cm mass was found peripherally in the right lobe of his liver, and an incisional biopsy was performed. Microscopic examination revealed a distinct overgrowth of clear cells in an acinar pattern, with tumor cells emerging directly from bile ducts. The tumor cells were periodic acid-Schiff reactive and diastase resistant, indicating the presence of mucin. No bile canaliculi were demonstrated by immunostaining with carcinoembryonic antigen. CT scans of the chest and abdomen were otherwise normal. Based on these microscopic, immunohistochemical, and clinical data, a diagnosis of clear cell cholangiocarcinoma was established. The patient later underwent reexploration and generous hepatic wedge resection. He did well postoperatively and is free of disease after 12 months. PMID- 10366217 TI - Spleen-preserving pancreatectomy for cystic pancreatic neoplasms. AB - Cystic neoplasms of the pancreas are an uncommon entity comprising fewer than 1 per cent of all pancreatic neoplasms. The guidelines for management of these tumors, specifically, the extent of resection, are unclear. Formerly, a distal pancreatectomy including the spleen was performed for tumors in the tail of the pancreas. The importance of preserving the spleen has been well documented; however, there are few reports of spleen-preserving pancreatectomy for cystic neoplasms of the distal pancreas. We report two patients who underwent spleen preserving pancreatectomy for mucinous cystic neoplasms in the tail of the pancreas. Both patients were female, ages 39 and 65 years. Preoperative preparation included administration of vaccinations and subcutaneous somatostatin. Operative technique emphasized division of the splenic artery and vein beyond the tip of the distal pancreas without mobilization of the spleen. The pancreas was transected with a vascular stapler. Fibrin glue was applied to the margin of the pancreas. The operative blood loss, duration of operation, and postoperative hospital stay were 150 and 250 mL, 150 and 180 minutes, and 7 and 9 days, respectively. The pathology revealed both lesions to be mucinous cystic neoplasms. The patients recovered and at 6-month follow-up were without complaints and in good health. Spleen-preserving pancreatectomy is rapid and associated with minimal morbidity. This procedure should be considered in the surgical management of cystic neoplasms in the tail of the pancreas. PMID- 10366218 TI - Quantitative sensation testing in epidemiological and therapeutic studies of peripheral neuropathy. PMID- 10366219 TI - Causes of the Gulf War syndrome: testing hypotheses. PMID- 10366220 TI - What does chronic electrical stimulation teach us about muscle plasticity? AB - The model of chronic low-frequency stimulation for the study of muscle plasticity was developed over 30 years ago. This protocol leads to a transformation of fast, fatigable muscles toward slower, fatigue-resistant ones. It involves qualitative and quantitative changes of all elements of the muscle fiber studied so far. The multitude of stimulation-induced changes makes it possible to establish the full adaptive potential of skeletal muscle. Both functional and structural alterations are caused by orchestrated exchanges of fast protein isoforms with their slow counterparts, as well as by altered levels of expression. This remodeling of the muscle fiber encompasses the major, myofibrillar proteins, membrane-bound and soluble proteins involved in Ca2+ dynamics, and mitochondrial and cytosolic enzymes of energy metabolism. Most transitions occur in a coordinated, time dependent manner and result from altered gene expression, including transcriptional and posttranscriptional processes. This review summarizes the advantages of chronic low-frequency stimulation for studying activity-induced changes in phenotype, and its potential for investigating regulatory mechanisms of gene expression. The potential clinical relevance or utility of the technique is also considered. PMID- 10366221 TI - Causalgia and reflex sympathetic dystrophy: does the sympathetic nervous system contribute to the generation of pain? AB - The striking response of causalgia and reflex sympathetic dystrophy (RSD) to sympatholytic procedures together with signs of autonomic nervous system abnormalities suggest that the sympathetic efferent system can generate or enhance pain (sympathetically maintained pain, SMP). This concept is supported by human and animal experiments indicating that sympathetic activity and catecholamines can activate primary afferent nociceptors. Some clinical evidence, however, calls the SMP concept into question and alternative explanations have been advanced. In this review, we describe the clinical features of causalgia and RSD and the evidence for sympatholytic efficacy. The major barrier to proving the SMP concept is that all available sympatholytic procedures are problematic. We conclude that, although the weight of current evidence supports the SMP concept and its relevance to causalgia and RSD, it remains unproven by scientific criteria. More careful adherence to diagnostic criteria and well-controlled trials of sympatholysis are needed to finally settle the issue. PMID- 10366222 TI - Effects of exposure to low-dose pyridostigmine on neuromuscular junctions in vitro. AB - During the Persian Gulf War, pyridostigmine bromide (PB), a reversible inhibitor of acetylcholinesterase, was used as prophylaxis against exposure to nerve gas. Exposure to PB has been suggested as a potential cause of the persistent fatigue reported among Gulf War veterans. The aim of this study was to evaluate the effects of acute and continuous exposure to low doses of PB on the neuromuscular junction. Organotypic spinal cord-muscle cocultures were used to examine in vitro the effects of PB under controlled conditions. Acute exposure to PB potentiated neuromuscular activity. Continuous exposure to PB produced a progressive decrease in the contractile activity of muscle fibers. Ultrastructural examination by electron microscopy revealed no abnormalities in the neuromuscular junctions after 1 week of exposure. Nerve terminal degeneration and atrophy of the postjunctional folds were evident after 2-week exposure to low-dose PB. The effects of PB were reversible following withdrawal. The reversibility of the PB induced changes in vitro suggests that such changes are causally unrelated to the fatigue reported by Persian Gulf War veterans years after exposure to PB. PMID- 10366223 TI - Muscle function in a patient with Brody's disease. AB - Adductor pollicis muscle function of a 21-year-old man with genetically confirmed Brody's disease (sarcoplasmic reticulum [SR] -Ca2+ATPase deficiency) was investigated to study the possible effects of reduced SR-Ca2+ATPase activity on muscle relaxation and force production. Following maximal electrical activation of the ulnar nerve, tetanic muscle half-relaxation time was greater in the patient (246 +/- 10 ms) than control subjects (97 +/- 4 ms, n = 8). During repetitive activation, there was a similar decline in maximal shortening velocity in the patient and controls, indicating a comparable reduction in cross-bridge cycling rate. The finding that the slowing of relaxation was greater in the patient (329 ms versus 138 +/- 20 ms) suggests that there was a further reduction of SR-Ca2+ATPase activity in the patient's muscle during fatigue. Following a voluntary contraction, involuntary activity of the antagonist muscles facilitated force decline and masked the impaired relaxation in the patient. This antagonist induced relaxation indicates that it might be difficult to establish impaired muscle relaxation with voluntary contractions. PMID- 10366224 TI - Polyneuropathy in autosomal dominant cerebellar ataxias: phenotype-genotype correlation. AB - Autosomal dominant cerebellar ataxias (ADCAs) are clinically and genetically heterogeneous neurodegenerative disorders. The aim of this study was to evaluate electrophysiologically peripheral nervous system involvement in each of the groups studied and its correlation with the number of CAG repeats. Forty patients with ADCA were clinically and electrophysiologically investigated. Thirty-five patients belonged to the ADCA type I group (SCA1, 12; SCA2, 10; SCA3, 13) and five to the ADCA type II group. Axonal sensory or sensorimotor polyneuropathy was found in 42% of the SCA1 patients, 80% of the SCA2 patients, and 54% of the SCA3 patients, whereas electrophysiological studies were normal in all those with ADCA type II. The number of CAG repeats was significantly higher in SCA1 patients with polyneuropathy than in those without polyneuropathy (P = 0.01), whereas the reverse was observed in SCA3/MJD (Machado-Joseph disease) patients (P = 0.05). We conclude that axonal polyneuropathy is often associated with ADCA type I, but its frequency varies according to factors such as the locus responsible and the number of CAG repeats. PMID- 10366225 TI - New near-nerve needle nerve conduction technique: differentiating epicondylar from cubital tunnel ulnar neuropathy. AB - At the elbow, the ulnar nerve is compressed most commonly either in the epicondylar groove or at the cubital tunnel. While conventional electrodiagnosis may localize an ulnar neuropathy to the elbow, separating epicondylar syndrome (tardy ulnar nerve palsy) from cubital tunnel syndrome is more difficult. We describe a new method using a near-nerve needle technique for distinguishing these two types of ulnar neuropathy at the elbow. We placed three active needle electrodes across the elbow: the first was 4 cm above, and the second and third were 1.5 cm and 6 cm below the medial epicondyle, respectively. The latter two points were chosen because of the presence of the cubital tunnel in this segment. Sensory, motor, and mixed nerve conduction studies (NCS) were performed on these two segments (elbow segment and cubital tunnel segment) in 26 normal nerves and normal data were established. We also present 7 cases of epicondylar ulnar nerve palsy and 1 case of cubital tunnel syndrome in which we were able to confirm the diagnosis with the present method. In 3 cases of epicondylar ulnar nerve palsy, the present method accurately localized the lesion when other methods failed. We believe that this method will be helpful in distinguishing cubital tunnel syndrome from epicondylar ulnar nerve palsy, especially in early ulnar neuropathy in which only sensory fibers are involved. PMID- 10366226 TI - Macrophages enhance muscle satellite cell proliferation and delay their differentiation. AB - This study investigated the effect of macrophages on in vitro satellite cell myogenesis in the turkey and mouse. Macrophages are considered to act as scavengers of tissue debris during the muscle degeneration-regeneration process. The number of dividing cells and of myoblasts expressing the myogenic regulatory factor MyoD indicated that macrophages enhanced satellite cell proliferation in both species. This was confirmed by observations with cultures treated for bromodeoxyuridine (BrdU) incorporation. In mouse and turkey macrophage-satellite cell cocultures, the number of differentiated myoblasts, the frequency of myogenin-positive cells, and the expression of developmental myosin isoforms were reduced as compared with control cultures, indicating that macrophages delayed satellite cell differentiation. The possibility that macrophages facilitate muscle fiber reconstitution by enhancing satellite cell proliferation should be taken into consideration in designing future strategies of satellite cell transplantation as a treatment for muscular dystrophies. PMID- 10366227 TI - Motor unit action potential components and physiologic duration. AB - Motor unit action potentials (MUAPs) recorded from the same motor unit at two distances along the biceps brachii muscle with monopolar needle electrodes at high amplifier gains (20 microV/division) and averaged 2000-3000 times reveal total potential durations of 39.6 +/- 4.6 ms. In addition, the terminal segment for each of these two MUAPs contained a late far-field potential with a mean duration of 23.8 +/- 4.1 ms. Computer simulations of MUAPs suggest that this long duration positive far-field mirrors the true morphology of the intracellular action potential (IAP), which is monophasic positive, possessing a terminal repolarization phase approaching 30 ms. This investigation suggests that the MUAP's physiologic duration is directly proportional to the muscle fiber length and the IAP's duration, which becomes manifest as a positive far-field potential when the IAP encounters the musculotendinous junction and slowly dissipates. The leading/trailing dipole model is used to explain qualitatively this study's quantitative clinical and computer simulation findings. PMID- 10366228 TI - Skeletal muscle and small-conductance calcium-activated potassium channels. AB - Skeletal muscle becomes hyperexcitable following denervation and when cultured in the absence of nerve cells. In these circumstances, the bee venom peptide toxin apamin, a blocker of small-conductance calcium-activated potassium (SK) channels, dramatically reduces the hyperexcitability. In this report, we show that SK3 channels are expressed in denervated skeletal muscle and in L6 cells. Action potentials evoked from normal innervated rat skeletal muscle did not exhibit an afterhyperpolarization, indicating a lack of SK channel activity; very low levels of apamin binding sites, SK3 protein, or SK3 mRNA were present. However, denervation resulted in apamin-sensitive afterhyperpolarizations and increased apamin binding sites, SK3 protein, and SK3 mRNA. Cultured rat L6 myoblasts and differentiated L6 myotubes contained similar levels of SK3 mRNA, although apamin sensitive SK currents and apamin binding sites were detected only following myotube differentiation. Therefore, different molecular mechanisms govern SK3 expression levels in denervated muscle compared with muscle cells differentiated in culture. PMID- 10366229 TI - Stimulation of peripheral nerves using a novel magnetic coil. AB - Magnetic nerve stimulation (MNS) using a novel figure-8 magnetic coil was compared with conventional electric nerve stimulation (ENS) in normal subjects and in patients with disorders of the peripheral nervous system. In contrast to previously tested coils, the virtual cathode of the novel coil was independent of the geometrical or electric conditions of the stimulated tissue. Maximal compound muscle action potentials (CMAPs) were elicited by MNS in all motor nerves tested. The slopes of the recruitment curves of ENS were steeper than those of MNS, indicating a comparatively lower maximal stimulation intensity and a higher intensity resolution of the magnetic stimulator. In four patients with entrapment syndromes at the ulnar groove, motor conduction velocities and amplitudes were similar for MNS and ENS across the affected nerve segment. However, in two patients with chronic inflammatory demyelinating polyneuropathy (CIDP), CMAPs were slightly smaller following MNS. This new technique is a promising step toward the ultimate goal of replacing ENS with MNS. PMID- 10366230 TI - Upper limb predominant, multifocal chronic inflammatory demyelinating polyneuropathy. AB - Chronic inflammatory demyelinating polyneuropathy (CIDP) presents in rare instances with focal or multifocal upper limb involvement. We reviewed the clinical and electromyographic (EMG) characteristics of 10 such patients (UL CIDP) and compared them with patients with typical generalized CIDP (G-CIDP) and multifocal motor neuropathy (MMN). There were six men and four women, with a mean age of 54 years. Symptoms began in one arm or hand in six patients and in both arms or hands in four and included numbness (n = 10), paresthesias (n = 9), weakness (n = 8), and pain (n = 6). Findings were initially restricted to the ulnar nerve distribution in three patients, and median and axillary nerve in one patient each, and involved multiple nerves in five. Conduction block was detected in the forearm segment of 68% of the median and ulnar motor nerves tested; in contrast to multifocal motor neuropathy, 73% of the sensory nerves tested were abnormal, and none had anti-GM1 antibodies. Aside from a regional onset, there were no clinical or electrophysiological features that distinguished patients with UL-CIDP from those with G-CIDP. However, the magnitude of recovery following treatment was greater in patients with G-CIDP. We conclude that a multifocal variant of CIDP begins with upper extremity sensorimotor symptoms, simulates isolated or multiple mononeuropathies, can be distinguished from MMN, and may have a less favorable response to treatment. PMID- 10366231 TI - Cervical root stimulation at C5/6 excites C8/T1 roots and minimizes pneumothorax risk. AB - Needle electrical cervical root stimulation may be performed lateral to the C5/C6 or C7/T1 spinous process interspaces. Pneumothorax has been reported following C7/T1 root stimulation. We evaluated the efficacy of a modified C5/C6 stimulation technique in exciting C8/T1 roots in 15 normal subjects and 36 patients with motor neuron disease (204 procedures). No instances of a 50% or greater amplitude decline occurred. C5/C6 interspace stimulation, therefore, may be used to excite C8/T1 roots while minimizing pneumothorax risk. PMID- 10366232 TI - Focal, steroid responsive myositis causing dropped head syndrome. AB - The dropped head syndrome, which occurs in a variety of neuromuscular disorders, is usually not due to an inflammatory process and generally either self-limited or nonresponsive to therapy. We present an 80-year-old woman who developed progressive neck weakness over a few months due to a focal and restricted inflammatory process involving the neck extensor muscles. She responded dramatically to treatment with immunosuppressive therapy. PMID- 10366233 TI - Suprascapular neuropathy related to a glenohumeral joint cyst. AB - A man with shoulder pain and complaints of weakness had examination findings consistent with a suprascapular neuropathy with predominant involvement of the infraspinatus muscle. Electrodiagnostic studies confirmed an axon-loss suprascapular neuropathy with greater involvement of the infraspinatus muscle. Magnetic resonance imaging (MRI) demonstrated a large ganglion cyst originating from the glenohumeral joint. The clinical, electrodiagnostic, and radiologic evaluation of suprascapular neuropathy is discussed. PMID- 10366234 TI - An unusual case of facial diplegia. PMID- 10366235 TI - Thoraconeuralgia gravidarum. PMID- 10366236 TI - On the classification of nonsimple motor unit potentials. PMID- 10366237 TI - Late-onset cytoplasmic body myopathy resembling myotonic dystrophy. PMID- 10366238 TI - 1999 Symposium on Advance Wound Care and Medical Research Forum on Wound Repair. Abstracts. PMID- 10366239 TI - The CPD Project. Continuing professional development. PMID- 10366240 TI - (S)-ketoprofen accumulation in premature neonates with renal failure who were exposed to the racemate during pregnancy. PMID- 10366241 TI - Serum concentrations and clinical effects of risperidone in schizophrenic patients in Singapore--a preliminary report. PMID- 10366242 TI - Strongyloides hyperinfection in adult T-cell leukaemia/lymphoma. PMID- 10366243 TI - Haemopoietic stem cell transplantation from unrelated donors. PMID- 10366244 TI - The history of haemophilia in the royal families of Europe. PMID- 10366245 TI - Successful autologous bone marrow transplantation in intravascular lymphomatosis. PMID- 10366246 TI - Activated protein C (APC) resistance in Indian children with thromboembolism. PMID- 10366247 TI - Pregnancy and aplastic anaemia. PMID- 10366248 TI - Fulminant hepatic failure after 2'-deoxycoformycin (pentostatin) PMID- 10366251 TI - Use of quantitative ASO-PCR to predict relapse in multiple myeloma. PMID- 10366249 TI - Maternal and neonatal autoimmune thrombocytopenia. PMID- 10366252 TI - Why discard the peritoneal macrophages of patients on CAPD? PMID- 10366253 TI - Investigation of minimal residual disease in childhood and adult acute lymphoblastic leukaemia by molecular analysis. PMID- 10366254 TI - Is filgrastim as useless after peripheral blood stem cell transplantation for adults as it could be for children? PMID- 10366255 TI - Intracytoplasmic detection of cytokines in neonatal lymphocytes and monocytes by flow cytometry. PMID- 10366256 TI - Healing of chronic leg ulcers in the hemoglobinopathies with perilesional injections of granulocyte-macrophage colony-stimulating factor. PMID- 10366257 TI - TT virus: a mother-to-child transmitted rather than bloodborne virus. PMID- 10366258 TI - [Study on effect of reinforcing kidney and invigorating spleen principle on severe asthma]. AB - OBJECTIVE: To explore on prevention and treatment of severe asthma with reinforcing the Kidney and invigoration the Spleen Principle (RKISP) and its mechanism. METHODS: Patients were treated with steroid aerosol (as control group), and both steroid aerosol and RKISP (as test group) respectively. RESULTS: The markedly effective rate of the test group was higher than that of the control group, the doses of beta-analeptic and the side-effect rates of the test group were lower than that of the control group, the pulmonary function of the test group was more remarkably improved. After treatment, the T cell hyperplasia response to specific allergen, the basophilic cell releasing capacity, the serum IgE and eosinophil of the test group were all reduced more obviously than those of the control group. In the control group, the neutrophil increased and the neutrophil phagocytic capability as well as the function of adrenal cortex were declined. While in the test group, the neutrophil phagocytic capability was raised, the neutrophil and the function of adrenal cortex had no significant changes. CONCLUSIONS: RKISP cooperated with steroid in relieving attack of asthma, and it was more important for it to display an obvious modulation action on immune and neuroendocrine system. PMID- 10366259 TI - Ethical dilemma: dealing with racist patients. Commentary: a role for personal values . . . and management. PMID- 10366260 TI - Ethical dilemma: dealing with racist patients. Commentary: isolate the problem. PMID- 10366261 TI - Ethical dilemma: dealing with racist patients. Commentary: courteous containment is not enough. PMID- 10366262 TI - Safety and effectiveness of nurse telephone consultation in out of hours primary care. Two interventions were combined as one. PMID- 10366263 TI - Clinical Trials. Randomised database studies could serve as new strategy. PMID- 10366264 TI - Prophylaxis against malaria. Preferred prophylaxis varies by region. PMID- 10366265 TI - Prophylaxis against malaria. More studies of mefloquine prophylaxis must be done in tourists. PMID- 10366266 TI - Cholesterol screening and management guidelines. Policy based on Sheffield table fully satisfies authors' criteria. PMID- 10366267 TI - People are "participants" in research. Many people are not "participants" in sense that this term was initially defined. PMID- 10366268 TI - People are "participants" in research. Pharmaceutical companies should follow medical profession's lead. PMID- 10366269 TI - Reporting on quality of life in RCTs. Authors are creating database of quality of life questionnaires. PMID- 10366270 TI - Values of educational visits in obstetrics. Good clinical audit is needed for interpreting systematic reviews. PMID- 10366271 TI - Values of educational visits in obstetrics. Staff changes will have affected ventouse rates. PMID- 10366273 TI - Underwater and Hyperbaric Medicine: from current literature. PMID- 10366272 TI - Medical student electives and infectious diseases. Risk of hepatitis C is greater than HIV. PMID- 10366274 TI - Isoforms of acyl carrier protein involved in seed-specific fatty acid synthesis. AB - Seeds of coriandrum sativum (coriander) and Thunbergia alata (black-eyed Susan vine) produce unusual monoenoic fatty acids which constitute over 80% of the total fatty acids of the seed oil. The initial step in the formation of these fatty acids is the desaturation of palmitoyl-ACP (acyl carrier protein) at the delta(4) or delta(6) positions to produce delta(4)-hexadecenoic acid (16:1(delta(4)) or delta(6)-hexadecenoic acid (16:1(delta(6)), respectively. The involvement of specific forms of ACP in the production of these novel monoenoic fatty acids was studied. ACPs were partially purified from endosperm of coriander and T. alata and used to generate 3H- and 14C-labelled palmitoyl-ACP substrates. In competition assays with labelled palmitoyl-ACP prepared from spinach (Spinacia oleracea), delta(4)-acyl-ACP desaturase activity was two- to threefold higher with coriander ACP than with spinach ACP. Similarly, the T. alata delta(6) desaturase favoured T. alata ACP over spinach ACP. A cDNA clone, Cs-ACP-1, encoding ACP was isolated from a coriander endosperm cDNA library. Cs-ACP-1 mRNA was predominantly expressed in endosperm rather than leaves. The Cs-ACP-1 mature protein was expressed in E. coli and comigrated on SDS-PAGE with the most abundant ACP expressed in endosperm tissues. In in vitro delta(4)-palmitoyl-ACP desaturase assays, the Cs-ACP-1 expressed from E. coli was four- and 10-fold more active than spinach ACP or E. coli ACP, respectively, in the synthesis of delta(4)-hexadecenoic acid from palmitoyl-ACP. In contrast, delta(9)-stearoyl-ACP desaturase activity from coriander endosperm did not discriminate strongly between different ACP species. These results indicate that individual ACP isoforms are specifically involved in the biosynthesis of unusual seed fatty acids and further suggest that expression of multiple ACP isoforms may participate in determining the products of fatty acid biosynthesis. PMID- 10366275 TI - Proceedings of the 11th International Symposium on Capillary Electroseparation Techniques. Venice, Italy, 4-7 October 1998. PMID- 10366276 TI - Proceedings of the Nidek International Excimer Laser Users Meeting. Cancun, Mexico, December 11-13, 1998. PMID- 10366277 TI - Post tonsillectomy pain. PMID- 10366278 TI - Retraction. PMID- 10366279 TI - Report from the 71st annual scientific meeting of the American Heart Association, Dallas, Texas, 9-11 November 1998. PMID- 10366280 TI - What we do not know about sequence analysis and sequence databases. PMID- 10366282 TI - Thoracic surgery and psychoneuro-immunology: brief history of a pioneer, Dr. T. Ishigami. PMID- 10366281 TI - Comparison of motion and stereopsis: linear and nonlinear performance. AB - To address the issue of whether the luminance-dependent (linear) and contrast dependent (nonlinear) processes in stereo and motion have a common computational basis, we compare both carrier-dependent and envelope-dependent performance for these two modalities by using the same stimulus and task: two-flash apparent motion/depth for a wide range of displacements. We do this for different densities, bandwidths, contrasts, spatial frequencies, and exposure durations. The results suggest that there is concordance not only between the luminance dependent (linear) processes of motion and stereo but also between the envelope dependent (nonlinear) processes of both modalities. Only one exception was found, but we show this to be amenable to an explanation based on a different contrast dependence for the nonlinear mechanisms of stereo and motion. This suggests that the computational basis of linear and nonlinear processes may be similar for stereopsis and motion. PMID- 10366283 TI - Ventricular septal defect and aortic regurgitation: have all the problems been elucidated? PMID- 10366285 TI - The black in American medicine. 1981. PMID- 10366284 TI - Spermatogonial stem cell transplantation, cryopreservation and culture. AB - Testis cells of a fertile male mouse can be transplanted to the seminiferous tubules of an infertile male, where the donor spermatogonial stem cells will establish spermatogenesis and produce spermatozoa that transmit the donor haplotype to progeny. In addition, stem cells can be cryopreserved for long periods, thereby making male germ lines immortal. Recently, mouse testis cells have been cultured for longer than 3 months and, following transplantation, produced spermatogenesis. These techniques are likely to be applicable to many species, since rat testis cells can be cryopreserved and generate spermatogenesis in the seminiferous tubules of immunodeficient mice. PMID- 10366286 TI - Literature reviews: adolescent medicine. PMID- 10366287 TI - Abstinence and safer sex HIV risk-reduction interventions for African-American adolescents. PMID- 10366288 TI - A genetic twist to the beta-agonist 'debate'. PMID- 10366289 TI - Human forearm venous occlusion plethysmography: methodology, presentation and analysis. PMID- 10366290 TI - Rejoinder to Bradley: Patient preferences and clinical trial design and appreciation and critique of a paper by Feine, Awad & Lund. PMID- 10366291 TI - "Non-alcoholic duct destructive chronic pancreatitis" or "primary chronic pancreatitis"? PMID- 10366292 TI - Guidelines for treatment of acute pancreatitis. PMID- 10366293 TI - Cholecystokinin provocation tests. PMID- 10366294 TI - Colonoscopic surveillance in ulcerative colitis. PMID- 10366295 TI - Requiem for the cholecystokinin provocation test. PMID- 10366296 TI - Cholecystokinin infusion and gall bladder dysfunction. PMID- 10366297 TI - Ductal dilatation and stenting for residual hepatolithiasis. PMID- 10366298 TI - The Helicobacter pylori breath test: a surrogate marker for peptic ulcer disease in dyspeptic patients. PMID- 10366299 TI - Can we ever be certain of the diagnosis of acute pancreatitis? PMID- 10366300 TI - MRCP and the diagnosis of suspected bile duct obstruction. PMID- 10366301 TI - Retrospective of a skin bank. PMID- 10366302 TI - Charles Baxter. From former ABA President David M. Heimbach, MD. PMID- 10366303 TI - The four milliliter man. PMID- 10366304 TI - Charles Baxter. From former ABA President Thomas L. Wachtel, MD, MMM. PMID- 10366305 TI - From the Nursing Forum editors: a tribute from Charlie's Angels. PMID- 10366306 TI - Issue dedicated to Charles R. Baxter, MD. PMID- 10366307 TI - Introduction. Lifetime achievements of Dr. Charles Rufus Baxter. PMID- 10366308 TI - More on the Pew report. PMID- 10366309 TI - Evidence-based dentistry. PMID- 10366310 TI - Stinging insect hypersensitivity: a practice parameter. The Joint Force on Practice Parameters, the American Academy of Allergy, Asthma and Immunology, the American College of Allergy, Asthma and Immunology, and the Joint Council of Allergy, Asthma and Immunology. PMID- 10366311 TI - Timing without a timer. PMID- 10366312 TI - Can a decay process explain the timing of conditioned responses? PMID- 10366313 TI - The whirligig of time: Some thoughts on Staddon and Higa. PMID- 10366314 TI - Time without clocks. PMID- 10366315 TI - Activities of plant-derived phenols in a fibroblast cell culture model. AB - Fourteen plant phenols of five structural groups (flavones, flavonols, flavan-3 ol, isoflavones, and phenylpropanoids) demonstrated concentration-dependent inhibitory or modulatory effects in a fibroblast cell culture model. The most potent inhibitory activity in this investigation was exhibited by apigenin (4), with flavone (1), chrysin (2), and genistein (9) being of somewhat lesser potency. These findings help to provide a better understanding of the action of these plant phenols in inflammatory/immune responses. PMID- 10366316 TI - Regarding "Saphenous surgery does not correct perforator incompetence in the presence of deep venous reflux". PMID- 10366317 TI - Regarding "Endothelium-dependent vasodilatation is impaired in both microcirculation and macrocirculation during acute hyperglycemia". PMID- 10366318 TI - Retraction: absence of human T-cell lymphotropic virus type I in cutaneous T-cell lymphoma. PMID- 10366319 TI - Prophylactic mastectomy in women with a high risk of breast cancer. PMID- 10366320 TI - Prophylactic mastectomy in women with a high risk of breast cancer. PMID- 10366321 TI - Prophylactic mastectomy in women with a high risk of breast cancer. PMID- 10366322 TI - Prophylactic mastectomy in women with a high risk of breast cancer. PMID- 10366323 TI - The one-in-nine risk of breast cancer. PMID- 10366324 TI - Hypovitaminosis D in a sunny country. PMID- 10366325 TI - Infected dog and cat bites. PMID- 10366326 TI - Infected dog and cat bites. PMID- 10366327 TI - Treatment of cat scratch disease. PMID- 10366328 TI - Treatment of cat scratch disease. PMID- 10366329 TI - Prosthetic-valve endocarditis caused by penicillin-resistant Streptococcus mitis. PMID- 10366330 TI - [Perinatal deaths in Delft and surrounds, 1983-1992: further reduction is possible by targeting lethal congenital abnormalities and placental insufficiency]. PMID- 10366331 TI - [Metamorphopsia of the Alice in Wonderland-syndrome]. PMID- 10366332 TI - Anti-inflammatory activity of phycocyanin extract in acetic acid-induced colitis in rats. AB - The anti-inflammatory effect of c-phycocyanin extract was studied in acetic acid induced colitis in rats. Phycocyanin (150, 200 and 300 mg kg(-1) p.o.) was administered 30 min gbefore induction of colitis with enema of 1 ml of 4% acetic acid per rat. Twenty-four hours later myeloperoxidase (MPO) activity was determined as well as histopathological and ultrastructural studies were carried out in colonic tissue. Phycocyanin substantially reduced MPO activity which was increase din the control colitis group. Also, histopathological and ultrastructural studies were carried out in colonic tissue. Phycocyanin substantially reduced MPO activity which was increased in the control colitis group. Also, histopathological and ultrastructural studies showed inhibition in inflammatory cell infiltration and reduction to some extent in colonic damage in rats treated with phycocyanin. The probable role of antioxidative and the scavenging properties of phycocyanin against reactive oxygen species in the anti colitic effect is discussed in this paper. To our knowledge this is the first report on the anti-inflammatory effect of phycocyanin in an experimental model of colitis. PMID- 10366333 TI - [Franco-Belgian consensus: gastroesophageal reflux in adults: "diagnosis and treatment". National French Society of Gastroenterology and Belgian Society of Gastroenterology]. PMID- 10366334 TI - [Osborn wave in hypothermia]. PMID- 10366335 TI - [Medical ethics guidelines for definition and determination of death with reference to organ transplantation. Swiss Academy of Medical Sciences]. PMID- 10366336 TI - Fertility therapy may aid gene transfer. PMID- 10366337 TI - Time cues help the brain see objects. PMID- 10366338 TI - AIDS now world's fourth biggest killer. PMID- 10366339 TI - HIV. French-led therapy fund kicks off in Africa. PMID- 10366340 TI - Anthropologists probe bones, stones and molecules. PMID- 10366341 TI - Threat to U.S. mouse colonies. PMID- 10366343 TI - Brain regions and drug addiction. PMID- 10366342 TI - Openness in private-public collaboration. PMID- 10366344 TI - Origin of the Japanese population. PMID- 10366345 TI - A fidgeter's calculation. PMID- 10366346 TI - Database protection: is it broken and should we fix it? PMID- 10366347 TI - GAD, a single autoantigen for diabetes. PMID- 10366348 TI - Nota bene: evolutionary biology. The male's dilemma. PMID- 10366349 TI - Findings in a fox rescued from hounds. PMID- 10366350 TI - "Omnicompetent" graduates. PMID- 10366351 TI - Charging for enforcement of controls on SRM. PMID- 10366352 TI - [Dissertations defended at the end of 1998-the start of 1999]. PMID- 10366353 TI - [The 75th anniversary of the Research Institute of Microbiology of the Ministry of Defense of the Russian Federation]. PMID- 10366354 TI - [A quarter of a century of the Central Medical Laboratory of the Air Force]. PMID- 10366355 TI - [The 1st Congress of Toxicologists of Russia]. PMID- 10366356 TI - The JAMA firing. PMID- 10366357 TI - Confessions of a beggarly doctor. PMID- 10366358 TI - Proceedings of the International Association for Surgical Metabolism and Nutrition Postgraduate Course on Nutrition. Acapulco, Mexico, August 24, 1997. PMID- 10366359 TI - The 8th Annual Congress of Japan Bi-Digital O-Ring Test Medical Society. Tokyo, Japan, July 19-20, 1998. Abstracts. PMID- 10366360 TI - 14th Annual International Symposium on Acupuncture and Electro-Therapeutics. New York City, New York, USA. October 22-25, 1998. Abstracts. PMID- 10366361 TI - Proceedings of the 1st Conference on Lipoxygenases. St. Julian, Malta, May 21-24, 1997. PMID- 10366362 TI - Economics and Cost-Effectiveness in Evaluating the Value of Cardiovascular Therapies. Proceedings of a symposium. St. Petersburg, Florida, USA. April 17-20, 1997. PMID- 10366363 TI - Orthodontics--as experienced by an adult. PMID- 10366364 TI - A worthy goal--less iatrogenics. PMID- 10366365 TI - History of Nephrology 3. Reports from the 2nd Congress of the International Association for the History of Nephrology. Padova, Italy. October 4-7, 1998. PMID- 10366366 TI - Amitriptyline use in geriatric care. PMID- 10366367 TI - [20th Alpine Symposium on Anatomy. Kotschach-Mauthen, 21-23 May 1998. Abstracts]. PMID- 10366368 TI - Low-molecular-weight heparins compared with unfractionated heparin for treatment of acute deep venous thrombosis. A cost-effectiveness analysis. AB - BACKGROUND: Low-molecular-weight heparins are effective for treating venous thrombosis, but their cost-effectiveness has not been rigorously assessed. OBJECTIVE: To evaluate the cost-effectiveness of low-molecular-weight heparins compared with unfractionated heparin for treatment of acute deep venous thrombosis. DESIGN: Decision model. DATA SOURCES: Probabilities for clinical outcomes were obtained from a meta-analysis of randomized trials. Cost estimates were derived from Medicare reimbursement and other sources. TARGET POPULATION: Two hypothetical cohorts of 60-year-old men with acute deep venous thrombosis. TIME HORIZON: Patient lifetime. PERSPECTIVE: Societal. INTERVENTION: Fixed-dose low-molecular-weight heparin or adjusted-dose unfractionated heparin. OUTCOME MEASURES: Costs, quality-adjusted life-years (QALYs), and incremental cost effectiveness ratios. An in-patient hospital setting was used for the base-case analysis. Secondary analyses examined outpatient treatment with low-molecular weight heparin. RESULTS OF BASE-CASE ANALYSIS: Total costs for inpatient treatment were $26,516 for low-molecular-weight heparin and $26,361 for unfractionated heparin. The cost of initial care was higher in patients who received low-molecular-weight heparin, but this was partly offset by reduced costs for early complications. Low-molecular-weight heparin treatment increased quality-adjusted life expectancy by approximately 0.02 years. The incremental cost-effectiveness of inpatient low-molecular-weight heparin treatment was $7820 per QALY gained. Treatment with low-molecular-weight heparin was cost saving when as few as 8% of patients were treated at home. RESULTS OF SENSITIVITY ANALYSIS: When late complications were assumed to occur 25% less frequently in patients who received unfractionated heparin, the incremental cost-effectiveness ratio increased to almost $75,000 per QALY gained. When late complications were assumed to occur 25% less frequently in patients who received low-molecular-weight heparin, this treatment resulted in a net cost savings. Inpatient low-molecular weight heparin treatment became cost saving when its pharmacy cost was reduced by 31% or more, when it reduced the yearly incidence of late complications by at least 7%, when as few as 8% of patients were treated entirely as outpatients, or when at least 13% of patients were eligible for early discharge. CONCLUSIONS: Low molecular-weight heparins are highly cost-effective for inpatient management of venous thrombosis. This treatment reduces costs when small numbers of patients are eligible for outpatient management. PMID- 10366369 TI - Low-molecular-weight heparins compared with unfractionated heparin for treatment of acute deep venous thrombosis. A meta-analysis of randomized, controlled trials. AB - BACKGROUND: Low-molecular-weight heparins may simplify the management of deep venous thrombosis. A critical clinical issue is whether this more convenient therapy is as safe and effective as treatment with unfractionated heparin. PURPOSE: To compare the safety and efficacy of low-molecular-weight heparins with those of unfractionated heparin for treatment of acute deep venous thrombosis. DATA SOURCES: Reviewers identified studies by searching MEDLINE, reviewing references from retrieved articles, scanning abstracts from conference proceedings, and contacting investigators and pharmaceutical companies. STUDY SELECTION: Randomized, controlled trials that compared a low-molecular-weight heparin preparation with unfractionated heparin for treatment of acute deep venous thrombosis. DATA EXTRACTION: Two reviewers extracted data independently. Reviewers evaluated study quality using a validated four-item instrument. DATA SYNTHESIS: Eleven of 37 studies met inclusion criteria for three major outcomes. Most studies used proper randomization procedures, but only one was double blinded. Compared with unfractionated heparin, low-molecular-weight heparins reduced mortality rates over 3 to 6 months of patient follow-up (odds ratio, 0.71 [95% CI, 0.53 to 0.94]; P = 0.02). For major bleeding complications, the odds ratio favored low-molecular-weight heparins (0.57 [CI, 0.33 to 0.99]; P = 0.047), but the absolute risk reduction was small and not statistically significant (0.61% [CI, -0.04% to 1.26%]; P = 0.07). For preventing thromboembolic recurrences, low-molecular-weight heparins seemed as effective as unfractionated heparin (odds ratio, 0.85 [CI, 0.63 to 1.14]; P > 0.2). CONCLUSIONS: Low molecular-weight heparin treatment reduces mortality rates after acute deep venous thrombosis. These drugs seem to be as safe as unfractionated heparin with respect to major bleeding complications and appear to be as effective in preventing thromboembolic recurrences. PMID- 10366370 TI - Cost-effectiveness of transesophageal echocardiography to determine the duration of therapy for intravascular catheter-associated Staphylococcus aureus bacteremia. AB - BACKGROUND: The appropriate duration of therapy for catheter-associated Staphylococcus aureus bacteremia is controversial. Conventional practice dictates that all patients receive prolonged courses of intravenous antibiotics. Some clinicians recommend abbreviated therapeutic courses, but an alternate approach involves prospectively identifying patients for whom abbreviated therapy is appropriate. OBJECTIVE: To determine the cost-effectiveness of transesophageal echocardiography (TEE) in establishing duration of therapy for catheter associated S. aureus bacteremia. DESIGN: Cost-effectiveness analysis. DATA SOURCES: MEDLINE search of literature; clinical data from patients with S. aureus bacteremia (n = 196) and patients with endocarditis (n = 60); and costs obtained from the study institution, regional home health agency, and national estimates of professional and technical fees. TARGET POPULATION: Patients with catheter associated S. aureus bacteremia on native heart valves without intravenous drug use or clinically apparent metastatic infection, immunosuppression, or indwelling prosthetic devices. TIME HORIZON: Patient lifetime. PERSPECTIVE: Societal. INTERVENTIONS: Antibiotic treatment based on TEE results compared with 2- or 4 week empirical therapy. OUTCOME MEASURES: Quality-adjusted life expectancy, costs, and incremental cost-effectiveness ratios. RESULTS OF BASE-CASE ANALYSIS: Compared with empirical short-course therapy, the TEE strategy cost $4938 per quality-adjusted life-year (QALY) gained. The effectiveness of the TEE strategy and the effectiveness of the long-course strategy were sufficiently similar that the additional cost of empirical long-course therapy ($1,667,971 per QALY) was higher than that which society usually considers cost-effective. RESULTS OF SENSITIVITY ANALYSES: In a four-way sensitivity analysis (endocarditis prevalence, TEE cost, short-course relapse rate, and TEE specificity), compared with empirical short-course therapy, the TEE strategy results ranged from cost savings to $155,624 per QALY. CONCLUSION: Within the limitations of existing empirical data, this study suggests that for patients with clinically uncomplicated catheter-associated S. aureus bacteremia, the use of TEE to determine therapy duration is a cost-effective alternative to 2- or 4-week empirical therapy. PMID- 10366371 TI - Long-term mortality after transsphenoidal surgery for Cushing disease. AB - BACKGROUND: Untreated Cushing disease historically has a high mortality rate, but the long-term survival of patients with Cushing disease after transsphenoidal surgery has not been reported. OBJECTIVE: To determine long-term mortality rate in patients who are treated for Cushing disease with current management techniques. DESIGN: Retrospective case series. SETTING: Tertiary care center. PATIENTS: 161 patients (32 men and 129 women; mean age, 38 years) who were treated for Cushing disease between 1978 and 1996. INTERVENTION: Transsphenoidal adenomectomy and as-needed adjunctive therapy. MEASUREMENT: Record review with follow-up interview. RESULTS: The cure rate for patients with microadenomas who had no previous therapy was 90% (123 of 137). No perioperative deaths occurred (0 of 193 procedures [95% CI, 0.0% to 1.9%]). Follow-up data (mean, 8.7 years) were obtained for 99% of patients (159 of 161). Six patients died. The 5- and 10-year survival rates were 99% (CI, 97% to 100%) and 93% (CI, 88% to 99%), respectively. Survival was similar to that seen in an age- and sex-matched sample that was based on U.S. population data (standardized mortality ratio, 0.98 [CI, 0.44 to 2.2]; P > 0.2). CONCLUSION: Survival of patients treated for Cushing disease with current management techniques between 1978 and 1996 was better than the poor survival historically associated with this disorder. PMID- 10366372 TI - Dialysis patients' preferences for family-based advance care planning. AB - BACKGROUND: Most patients do not participate in advance care planning with physicians. OBJECTIVE: To examine patients' preferences for involving their physicians and families in advance care planning. DESIGN: Face-to-face interviews with randomly selected patients. SETTING: Community-based dialysis units in one rural and one urban region. PARTICIPANTS: 400 hemodialysis patients. MEASUREMENTS: Questions about whom patients involve in advance care planning, whom patients would like to include in this planning, and patients' reactions to state legislation on surrogate decision makers in end-of-life care. RESULTS: Patients more frequently discussed preferences for end-of-life care with family members than with physicians (50% compared with 6%; P < 0.001). More patients wanted to include family members in future discussions of advance care planning than wanted to include physicians (91% compared with 36%; P < 0.001). Patients were most comfortable with legislation that granted their family end-of-life decision-making authority in the event of their own incapacity (P < 0.001). CONCLUSION: Most patients want to include their families more than their physicians in advance care planning. PMID- 10366373 TI - Cross-cultural primary care: a patient-based approach. AB - In today's multicultural society, assuring quality health care for all persons requires that physicians understand how each patient's sociocultural background affects his or her health beliefs and behaviors. Cross-cultural curricula have been developed to address these issues but are not widely used in medical education. Many curricula take a categorical and potentially stereotypic approach to "cultural competence" that weds patients of certain cultures to a set of specific, unifying characteristics. In addition, curricula frequently overlook the importance of social factors on the cross-cultural encounter. This paper discusses a patient-based cross-cultural curriculum for residents and medical students that teaches a framework for analysis of the individual patient's social context and cultural health beliefs and behaviors. The curriculum consists of five thematic units taught in four 2-hour sessions. The goal is to help physicians avoid cultural generalizations while improving their ability to understand, communicate with, and care for patients from diverse backgrounds. PMID- 10366374 TI - A consensus-based approach to providing palliative care to patients who lack decision-making capacity. ACP-ASIM End-of-Life Care Consensus Panel. American College of Physicians-American Society of Internal Medicine. AB - Making palliative care decisions for a patient who lacks decision-making capacity presents several challenges. Other people, such as family and caregivers, must choose for the patient. The goals and values of these decision makers may conflict with those of each other and with those of the patient, who now lacks the capacity to participate in the decision. This paper presents a case study of a patient with severe Alzheimer disease who has two common clinical problems: neurogenic dysphagia and aspiration pneumonia. The case study describes a consensus-based decision-making strategy that keeps what is known about the patient's wishes and values in the foreground but also expects guidance from the physician and elicits input from family members and other people who care for and have knowledge about the patient. The steps of this process, including key clinical prompts and potential transition statements, are outlined and described. The overall goal of the case commentary is to demonstrate that physicians can guide a highly emotional and personal process in a structured manner that has meaning for the patient, family, physician, and other caregivers. PMID- 10366375 TI - Primary angioplasty compared with thrombolysis: new issues in the era of glycoprotein IIb/IIIa inhibition and intracoronary stenting. AB - The past decade has witnessed a dramatic expansion in the scope of both mechanical and pharmacologic methods for opening occluded arteries in patients with acute myocardial infarction. Although the relative merits of conventional balloon angioplasty and thrombolysis have been evaluated, this old debate is being eclipsed by new comparisons. New device technologies, such as intracoronary stenting; more potent and more fibrin-specific thrombolytic agents; and new antithrombotic and antiplatelet agents all offer the potential for improved outcomes. But despite these recent developments, the time-dependent open artery hypothesis--which states that the achievement of early, full, and sustained reperfusion is associated with better outcomes--remains essentially unchanged. This article reviews data on the ability of six revascularization strategies- stand-alone thrombolysis, conventional percutaneous transluminal coronary angioplasty, stenting, glycoprotein IIb/IIIa inhibitors plus thrombolytic agents, glycoprotein IIb/IIIa inhibitors plus interventions, and the combination of pharmacologic and mechanical interventions--to produce early, full, and sustained reperfusion. PMID- 10366376 TI - The evolving role of ambulatory arrhythmia monitoring in general clinical practice. AB - PURPOSE: To evaluate the efficacy of various ambulatory electrocardiographic monitors for the diagnosis of arrhythmia-related disorders and to provide recommendations for their use in clinical practice. DATA SOURCES: Studies published since 1988 were identified through search of the MEDLINE database. STUDY SELECTION: Studies that met methodologic criteria for minimal bias and had clinical relevance were selected. DATA EXTRACTION: Descriptive and analytic data from each study. DATA SYNTHESIS: Ambulatory electrocardiographic monitors, specifically transtelephonic continuous-loop event recorders, are highly effective for establishing a diagnosis in patients with palpitations but are less effective for establishing a diagnosis in patients with syncope. Clinicians may use these devices to monitor for nonsustained ventricular tachycardia in patients at potentially high risk for sudden arrhythmic death; however, few data are available to support this practice. Ambulatory monitors are useful for assessment of the safety and efficacy of antiarrhythmic medications and the recurrence of symptomatic supraventricular arrhythmias. New ambulatory arrhythmia monitoring devices are being developed that may facilitate outpatient management of chronic cardiac disease. CONCLUSIONS: Ambulatory electrocardiographic monitors, particularly transtelephonic continuous-loop event recorders, aid in the diagnosis of symptomatic arrhythmias. These devices are also useful for monitoring the effectiveness and safety of antiarrhythmic medications. Guidelines are lacking for use of these devices to assess prognosis in patients at potential risk for sudden arrhythmic death. PMID- 10366377 TI - High-priced technology can be good value for money. PMID- 10366378 TI - The doctor in the family. PMID- 10366379 TI - American College of Rheumatology criteria for the diagnosis of vasculitis. PMID- 10366380 TI - American College of Rheumatology Criteria for the diagnosis of vasculitis. PMID- 10366381 TI - Diabetes case management. PMID- 10366382 TI - Diabetes case management. PMID- 10366383 TI - Diabetes case management. PMID- 10366384 TI - Perioperative supraventricular tachyarrhythmia. PMID- 10366385 TI - Therapy for Legionnaires disease. PMID- 10366386 TI - Nutrition and policy. 2: The role of professional societies. PMID- 10366387 TI - Point-of-care testing: instant gratification? PMID- 10366388 TI - [Microsurgery and the 22nd congress of GAM (Group of Advancement of Microsurgery). Proceedings. Paris, France, April 30-May 1, 1999]. PMID- 10366389 TI - The power of public relations. PMID- 10366390 TI - Needs survey of Canadian rosacea patients. AB - BACKGROUND: In 1995, a Rosacea Awareness Program (RAP) was initiated in Canada to make available educational resources for physicians and rosacea patients. Material is in French or English, and is accessible though physician offices and by toll-free telephone. Information was communicated to the public via noncommercial, editorial media. The RAP created a database of rosacea patients in Canada. OBJECTIVE: We investigated if individuals in the database had a confirmed diagnosis of rosacea, how they perceived their treatment by the medical system, and identified their needs. METHODS: A two-page questionnaire was mailed to 7874 individuals registered with the RAP. Thirty percent of these individuals responded. Where comparisons were made a chi-squared statistic was used. RESULTS: Over 70% learned of the RAP via public media. It took patients an average of 5 years to have a diagnosis made after the first symptoms appeared. In the majority of patients (53%) the diagnosis was ultimately made by a specialist. Fifty-eight individuals said they had not discussed their condition with their doctor. Patients were likely to continue on medication that was prescribed (60%) and topical metronidazole was the most common medication used, mostly the gel formulation. Most patients used these twice daily. Patients were very satisfied with treatments and almost 90% had reduced symptoms. Despite receiving explanations and written material, patients expressed a strong interest in more information being available on skin care, make-up, and psychological aspects of rosacea. CONCLUSIONS: The RAP provides a needed educational service and is a useful database. Patients are very knowledgeable about their disease, but despite this and excellent therapeutic responses, the patients demand more information. PMID- 10366392 TI - Monetary and nonmonetary costs to patients attending an ambulatory dermatology clinic. AB - BACKGROUND: Despite universal coverage under a provincial health plan, the residents of Ontario, Canada, still bear some costs for outpatient care, particularly for prescription drugs. OBJECTIVE: To determine the financial and nonmonetary costs borne by patients presenting at a dermatology clinic in an academic centre, and to assess the extent to which these costs were problematic. METHODS: Consecutive new patients in a 6-week period completed a self administered questionnaire. RESULTS: Eighty-six of 140 questionnaires (61%) were returned for analysis. The mean total cost to patients was C$28.92 (range $0 to $177.00). Medications were the largest expense (mean $35.66 for those receiving medication). Despite relatively prompt referrals (mean 12.4 days) and short in office waiting time (mean 26.5 minutes), there was a trend for subjects to rate time costs as more problematic than monetary costs. CONCLUSION: Patients attending a dermatology clinic bear variable monetary and nonmonetary costs. For some patients these costs may have the potential to impair access to care. PMID- 10366391 TI - Impact of a sun awareness curriculum on medical students' knowledge, attitudes, and behaviour. AB - BACKGROUND: Physicians teach sun awareness to their patients, but frequently have no formal training in this area. A week-long dermatology curriculum during Sun Awareness Week that included skin cancer and sun awareness education to first year medical students was introduced in May 1998 at the University of Western Ontario, London, Canada. OBJECTIVE: The purpose of this study was to determine the baseline knowledge, attitudes, and behaviour of the first-year medical students towards sun awareness before and after the new curriculum. METHOD: This study used a pre- and post-test design to determine the impact of the curriculum on the medical students' knowledge, attitudes, and intent to change behaviour. It also reports any influence of demographic variables on these parameters. RESULTS: The students demonstrated a substantial improvement in their knowledge of sun related topics despite some baseline knowledge. Many students reported unhealthy behaviour prior to the curriculum, but demonstrated an intent to adopt more healthy behaviour after the curriculum. Minor differences in knowledge and behaviour due to demographic characteristics disappeared upon completion of the curriculum. CONCLUSIONS: An undergraduate medical curriculum with skin cancer and sun awareness education can improve the medical students' knowledge, attitudes, and behaviour towards sun awareness. PMID- 10366393 TI - Rapidly growing squamous cell carcinoma. AB - BACKGROUND: Squamous cell carcinoma (SCC) may present with a history of rapid growth. Although multiple subtypes have been described regarding histologic characteristics and etiology, the subset of rapidly growing squamous cell carcinomas (RGSCC) has not been described. OBJECTIVE: To evaluate and describe the clinical and histologic characteristics of squamous cell carcinomas that grow rapidly. METHODS: Recorded clinical data and biopsies of 26 lesions with a history of rapid growth and histologically diagnosed as SCC were reviewed. RESULTS: Rapidly growing SCC occurred most commonly on the head and neck, followed by hands and extremities, and had an average duration of 7 weeks before diagnosis. The average size of the lesions was 1.29 cm and nearly 20% occurred in immunosuppressed patients. CONCLUSIONS: Some SCCs may grow rapidly. The reason for the rapid growth is not clear and several hypotheses are discussed including immunosuppression and viral etiology. These lesions should be treated aggressively as their behaviour and prognosis are not yet well described. PMID- 10366394 TI - Point-counterpoint: "Allergy" in atopic dermatitis. PMID- 10366395 TI - Point-counterpoint: "Allergy in atopic dermatitis: more harm than good?". PMID- 10366396 TI - A patient with a refractory cutaneous bullous eruption. PMID- 10366397 TI - Certain gut polyposis syndromes of importance: summary notes. PMID- 10366398 TI - The "sins" of the fathers: self-healing squamous epithelioma in Scotland. AB - BACKGROUND: A review is presented of the recent progress made in mapping of the hereditary skin disease "Ferguson-Smith multiple self-healing squamous epithelioma." METHODS: The use of founder effects in an autosomal dominant disease is reviewed as applied to gene mapping efforts. RESULTS: A common haplotype among Scottish families segregating Ferguson-Smith disease allowed the narrowing of the candidate gene interval and the identification of several possible disease-associated genes. CONCLUSION: The gene for Ferguson-Smith multiple self-healing squamous epithelioma lies in a narrow region on chromosome 9q, along with several other important hereditary skin disease loci. PMID- 10366399 TI - Increased soft tissue in the posterior cervical and upper back area of patients on HIV-1 protease inhibitors. AB - BACKGROUND: Corticosteroids as well as sex hormones affect the redistribution of subcutaneous fat and the percentage of lean body mass. In addition, some stromal cells express steroid receptors, and the quantity and distribution of these receptors vary at different body sites and between sexes. Inhibitors of HIV-1 protease may affect steroid hormone metabolism through their effect on cytochrome P450. OBJECTIVES: To determine the changes in the tissue of the back in three HIV 1+ patients who developed increased soft tissue in posterior cervical and upper back areas while on HIV-1 protease inhibitors. METHODS: Punch biopsies of the involved posterior cervical and upper back areas were done. These included subcutaneous adipose tissue. Routine hematoxylin and eosin-stained sections, along with special stains for elastic and stromal mucin, and immunohistochemical stains for CD34 (HPCA-1 and Factor XIIIa) were evaluated. RESULTS: Histologically all three patients showed identical features. There was expansion of the dermis with decreased periadnexal fat and marked widening of the fibrous septa within the expanded subcutaneous fat. CONCLUSIONS: The posterior cervical and upper back area appears to be a common site for localization of mesenchymal tumours that show some fat differentiation and produce an increase in stromal matrix material. Mesenchymal cell populations within this area are also affected by systemic diseases. A male predominance pattern occurs with these conditions, and steroid receptors are expressed on some mesenchymal cells, that vary with the body location. Thus, this observation may be related to the effects of protease inhibitors on steroid hormone metabolism through their inhibition of cytochrome P 450. PMID- 10366400 TI - Mycobacterium marinum with associated bursitis. AB - BACKGROUND: Mycobacterium marinum infections have been reported for over 50 years, mostly in association with trauma in the setting of water exposure. OBJECTIVE: The differential diagnosis for nodules in a sporotrichoid distribution with simultaneous bursitis is discussed. Mycobacterium marinum treatment regimens for skin and joint involvement are reviewed. METHODS: Mycobacterium marinum was identified by skin tissue culture with Lowenstein-Jensen medium at 32 degrees C. Histopathologic findings support mycobacterial infection. RESULTS: Bursitis and nodules resolved in the first 2 months of a 6-month course of minocycline treatment. CONCLUSION: Bursitis is an extremely rare but significant complication of M. marinum. PMID- 10366401 TI - Chronic urticaria: review of the literature. PMID- 10366402 TI - Exposure to exogenous estrogens in food: possible impact on human development and health. AB - There has been increasing concern about the impact of environmental compounds with hormone-like action on human development and reproductive health over the past decades. An alternative but neglected source of hormone action that may be considered in this connection is hormone residues in meat from husbandry animals treated with sex steroid hormones for growth promotion. Treatment of cattle with naturally occurring or synthetic sex hormones may enhance lean muscle growth and improve feed efficiency and is therefore a very cost effective procedure for cattle producers who have used it for decades in some Western countries, including the USA and Canada. The Joint Food and Agricultural Organisation/World Health Organisation (FAO/WHO) expert committee on food additives (JECFA) and the US Food and Drug Administration (FDA) considered, in 1988, that the residues found in meat from treated animals were safe for the consumers. We have re evaluated the JECFA conclusions regarding the safety of estradiol residues in meat in the light of recent scientific data, with special emphasis on estradiol levels in prepubertal children. These levels are needed for estimates of the normal daily production rates of estradiol in children, who may be particularly sensitive to low levels of estradiol. In our opinion, the conclusions by JECFA concerning the safety of hormone residues in meat seem to be based on uncertain assumptions and inadequate scientific data. Our concerns can be summarized as follows. 1) The data on residue levels in meat were based on studies performed in the 1970's and 1980's using radioimmunoassay (RIA) methods available at the time. The sensitivity of the methods was generally inadequate to measure precisely the low levels found in animal tissues, and considerable variation between different RIA methods for measuring steroids exists. Therefore the reported residue levels may be subject to considerable uncertainty. 2) Only limited information on the levels of the various metabolites of the steroids was given despite the fact that metabolites also may have biological activity. 3) Reliable data on daily production rates of steroid hormones were and are still lacking in healthy prepubertal children. This lack is crucial as previous guidelines regarding acceptable levels of steroid residues in edible animal tissues have been based on very questionable estimates of production rates in children. Thus, even today the US FDA bases its guidelines on the presumably highly overestimated production rates in prepubertal children given in the JECFA 1988 report. 4) The possible biological significance of very low levels of estradiol is neglected. In conclusion, based on our current knowledge possible adverse effects on human health by consumption of meat from hormone-treated animals cannot be excluded. PMID- 10366403 TI - What do we call a goiter? PMID- 10366404 TI - Local versus WHO/International Council for Control of Iodine Deficiency Disorders recommended thyroid volume reference in the assessment of iodine deficiency disorders. AB - OBJECTIVE: Iodine deficiency endemia is defined by the goitre prevalence and the median urinary iodine concentration in a population. Lack of local thyroid volume reference data may bring many health workers to use the European-based WHO/International Council for Control of Iodine Deficiency Disorders (ICCIDD) recommended reference for the assessment of goitre prevalence in children in different developing countries. The present study was conducted in non-iodine deficient areas in Malaysia to obtain local children's normative thyroid volume reference data, and to compare their usefulness with those of the WHO/ICCIDD recommended reference for the assessment of iodine-deficiency disorders (IDD) in Malaysia. DESIGN AND METHODS: Cross-sectional thyroid ultrasonographic data of 7410 school children (4004 boys, 3406 girls), aged 7-10 years, from non-iodine deficient areas (urban and rural) in Peninsular Malaysia were collected. Age/sex- and body surface area/sex-specific upper limits (97th percentile) of normal thyroid volume were derived. Thyroid ultrasonographic data of similar-age children from schools located in a mildly iodine-deficient area, a severely iodine-deficient area, and a non-iodine-deficient area were also collected; spot urines were obtained from these children for iodine determination. RESULTS: The goitre prevalences obtained using the local reference were consistent with the median urinary iodine concentrations in indicating the severity of IDD in the areas studied. In contrast, the results obtained using the WHO/ICCIDD-recommended reference showed lack of congruency with the median urinary iodine concentrations, and grossly underestimated the problem. The local sex-specific reference values at different ages and body surface areas are not a constant proportion of the WHO/ICCIDD-recommended reference. A further limitation of the WHO/ICCIDD-recommended reference is the lack of normative values for children with small body surface areas (<0.8m2) commonly found in the developing countries. CONCLUSION: The observations favour the use of a local reference in the screening of children for thyroid enlargement. PMID- 10366405 TI - Thyroid volumes in US and Bangladeshi schoolchildren: comparison with European schoolchildren. AB - OBJECTIVE: The World Health Organization (WHO) recently adopted thyroid volume ultrasonography results from European schoolchildren as the international reference for assessing iodine deficiency disorders. Our objective was to describe thyroid volumes measured by ultrasonography in US and Bangladeshi schoolchildren and compare these with European schoolchildren. METHODS: Cross sectional studies were performed in schoolchildren in the US (n=302) and Bangladesh (n=398). Data were collected on the following: thyroid size by palpation and ultrasonography; urinary iodine; age; sex; weight; and height. RESULTS: Applying the new WHO thyroid volume references to the Bangladeshi children resulted in prevalence estimates of enlarged thyroid of 26% based on body surface area (BSA) and 7% based on age. In contrast, in the US children, the prevalence estimates were less than 1% for each reference. In the US children, the best single predictor of thyroid volume was BSA (R2=0.32), followed by weight (R2=0.31). Using linear regression, upper normal limits (97th percentile) of thyroid volume from US children were calculated for BSA, weight and age, and were found to be lower than the corresponding references based on BSA and age from European schoolchildren. CONCLUSIONS: In areas with malnutrition, such as Bangladesh, the BSA reference should be preferred to the reference based on age. Results from the US children indicated that a thyroid volume reference based on weight alone would perform as well as the one based on BSA. European schoolchildren had larger thyroids than US children, perhaps due to a residual effect of iodine deficiency in the recent past in some areas in Europe. PMID- 10366406 TI - The predominant form of non-toxic goiter in Greece is now autoimmune thyroiditis. AB - Endemic non-toxic goiter (NTG) in Greece has been attributed primarily to iodine deficiency. Thirty years ago about 60% of the prepubertal boys and girls examined in endemic goiter regions presented with NTG and among them thyroid autoimmunity was rarely detected. Although iodine supplementation has corrected this deficiency during the past 30 years, new cases of NTG still appear. To evaluate the prevalence and type of NTG and the effect of iodine supplementation on them in Greece at present, we performed two cross-sectional clinical studies and a retrospective pathology one: (i) thyroid gland volume and urinary iodine excretion (UIE) were assessed in a representative sample of 1213 schoolchildren from previously endemic and non-endemic regions; (ii) serum thyroxine, tri iodothyronine, TSH, thyroid autoantibodies (AAB) (anti-thyroid peroxidase and anti-thyroglobulin antibodies) and UIE (in 60 patients) were measured in 300 consecutive patients with NTG from Athens and Heraklion; and (iii) we compared the prevalence of autoimmunity among fine needle aspiration smears of benign thyroid pathologies performed by the same pathologist between 1985 and 1986 (975 cases) and between 1994 and 1995 (2702 cases). We found that 12. 5% of the schoolchildren examined in regions with a previous history of endemic goiter had NTG, whereas this percentage was only 1.7% in areas without such a history. In Athens (61.6%) and Heraklion (58. 5%) a substantial number of NTG patients were AAB positive and biochemically hypothyroid. UIE in Athens did not differ between patients with autoimmune goiter (ATG) and simple goiter. The prevalence of autoimmune stigmata in pathology smears has increased from 5.94% (years 1985 1986) to 13.91% (years 1994-1995) (P<0.05). We conclude that: (i) the persistence of endemic goiter in regional foci despite iodine deficiency correction suggests a possible role for a naturally occurring goitrogen; (ii) ATG is the predominant form of NTG in Greece nowadays; and (iii) the five-fold decrease in the prevalence of NTG during the past 30 years followed by the increase of ATG may support the relative character of the latter. PMID- 10366407 TI - Thyroid function, morphology and autoimmunity in young patients with insulin dependent diabetes mellitus. AB - OBJECTIVE: An association between insulin-dependent diabetes mellitus (IDDM) and autoimmune thyroid disease is well recognized. We have studied the prevalence of thyroid dysfunction, autoimmunity and morphological abnormalities by ultrasonography in young diabetics. SUBJECTS AND METHODS: Among young IDDM patients less than 18 years old and living in the county of Funen, Denmark, 105 of 116 eligible patients participated. They were compared with 105 healthy children matched for sex and age. Routine thyroid function parameters (thyroxine (T4), tri-iodothyronine (T3), T3 resin uptake and TSH) and thyroid autoantibodies (anti-thyroid peroxidase, TPOab, and thyroglobulin antibodies, Tgab) were measured. Thyroid size and morphology were determined by ultrasonography. RESULTS: Two of the diabetics had previously diagnosed hypothyroidism and three new cases of subclinical hypothyroidism were found. There were no significant differences in thyroid function variables or thyroid volume between diabetics and controls. Thyroid volume correlated significantly with age and weight in both groups. Among diabetics, 17 had thyroid autoantibodies (13 with TPOab, 14 with Tgab and 10 with both) compared with 2 children in the control group (P<0.001). Forty-four with IDDM as opposed to 11 of the controls (P<0.001) had morphological abnormalities at ultrasonography. Most of them had various degrees of hypoechogenicity thought to be a marker of thyroid autoimmunity. Among the 17 diabetics with autoantibodies, 10 had morphological abnormalities at ultrasonography. CONCLUSIONS: A high proportion of young IDDM patients without any clinical signs of thyroid disease have markers of thyroid autoimmunity. Many have thyroid autoantibodies, but even more have abnormalities by thyroid ultrasonography. PMID- 10366408 TI - Mono- and plurihormonal thyrotropic pituitary adenomas: pathological, hormonal and clinical studies in 12 patients. AB - In a series of 12 patients (eight women and four men, aged between 20 and 62 years), operated on for a pituitary adenoma shown to be thyrotropic by immunocytochemistry, we performed a retrospective and comparative analysis of clinical and biological data, tumor studies including immunocytochemistry with double labeling, and proliferation marker (proliferative cell nuclear antigen (PCNA) and Ki-67) detection, electron microscopy and culture. Our study leads us to confirm that thyrotropic tumors are rare (12 of 1174 pituitary adenomas: 1%). The main points arising were that: (1) high or normal plasma TSH associated with an increase in plasma alpha-subunit and high thyroid hormone levels is the best criterion for diagnosis; (2) the failure of TSH to respond to TRH or Werner's test is not a reliable criterion for diagnosis; (3) thyrotropic adenomas may be 'silent', without clinical signs of hyperthyroidism and with only slight increase in TSH, tri-iodothyronine and thyroxine concentrations; (4) mitoses and nuclear atypies are frequently detected in large tumors, which are invasive in more than 50% of cases - the first analysis of two proliferation markers (PCNA and Ki-67) bears out the relative aggressiveness of thyrotropic adenomas; (5) thyrotropic adenomas are frequently plurihormonal. Immunocytochemical double labeling, complemented by in vitro study, showed that thyrotropic tumor cells sometimes can or sometimes cannot cosecrete TSH, GH or prolactin. The pathological identification of monohormonal and plurihormonal adenomas seems to be supported by clinical and biological differences. PMID- 10366409 TI - Clinical and morphological features of undifferentiated monomorphous GH/TSH secreting pituitary adenoma. AB - A 41-year-old male presented with progressive visual defects, acromegaly and hyperthyroidism. After clinical evaluation a giant GH/TSH-secreting pituitary adenoma was diagnosed. Administration of the somatostatin analog octreotide at doses of 150 microg s.c. per day inhibited the secretion of both GH and TSH. A three-week treatment with octreotide prior to surgery led to slight visual improvement and CT scan showed some new necrotic areas within the tumor mass. Transcranial surgery was performed. By immunohistochemical analyses of the adenoma tissue GH, prolactin and beta-chorionic gonadotropin were detected; TSH was negative. Electron microscopy revealed an undifferentiated, monomorphous adenoma with morphological features of an acidophil stem cell adenoma such as the presence of misplaced exocytoses, fibrous bodies and mitochondrial gigantism. However, the tumor cells contained small secretory granules (up to 250 nm) accumulated along the cell membrane characteristic of thyrotrope cells. Furthermore, some adenoma cells were fusiform with long cytoplasmic processes resembling thyrotropes. Two months after the operation CT scan revealed a large residual tumor. Serum GH and TSH levels had increased again and the TSH level was even higher than before the treatment. The patient died suddenly, most probably of lethal arrhythmia. Specimens of the adenoma tissue obtained at autopsy confirmed the previous findings with the exception of positive immunostaining for TSH which was found in less than 1% of the adenoma cells. This undifferentiated, monomorphous GH/TSH-secreting pituitary adenoma represents an entity that is unusual both in its ultrastructural features and clinical manifestations suggesting a cytogenesis from an early, undifferentiated stem cell. PMID- 10366410 TI - Urinary gonadotropin peptide as acute phase reactant: transient elevation after operation for digestive diseases. AB - OBJECTIVE: In order to characterize urinary gonadotropin peptide (UGP) as an acute phase reactant, we focused on the UGP levels after surgical operation. DESIGN: Fifty cases of gastrointestinal cancer, 4 cases of cancers of other organs and 13 cases of benign digestive diseases were enrolled into this study. METHODS: UGP levels were measured using an enzyme immunoassay before and after surgery. RESULTS: Fifty-four (80.6%) of the 67 cases studied showed transient elevations of UGP. Both urinary interleukin (IL)-6 and LH levels were also increased transiently in 49 cases (73.1%). All of these three factors were increased in 38 cases (56.7%), and in 32 (84.2%) of these 38 cases, the order of peak appearance was as follows: IL-6, LH, and UGP. The UGP levels in the group of total gastrectomy were significantly higher than those in the group of partial gastrectomy. CONCLUSIONS: These results suggest that UGP shows a transient peak after surgery, correlating with levels of cytokines such as IL-6. UGP may be an acute phase reactant, and its levels are correlated with the grade of surgical stress. PMID- 10366412 TI - Somatic MEN1 gene mutation does not contribute significantly to sporadic pituitary tumorigenesis. AB - Pituitary adenomas are a common manifestation of multiple endocrine neoplasia type 1 (MEN1) but most of them occur sporadically. There are only a few well defined genetic abnormalities known to occur in these sporadic tumours. The MEN1 gene located on 11q13 has recently been cloned and allelic deletion and mutation analysis studies have implicated the MEN1 gene in a significant fraction of the sporadic counterparts of typical MEN1 neoplasms (parathyroid tumours, insulinomas and gastrinomas). To determine if MEN1 gene inactivation is also involved in the development of sporadic pituitary adenomas, allelic deletions of chromosome 11q13 and MEN1 gene mutations and polymorphisms were assessed in 35 sporadic tumours of the anterior pituitary (9 prolactin-secreting, 8 GH-secreting, 3 TSH-secreting, 2 TSH/GH-secreting, 4 Cushing, 9 silent). Thirty-one tumours were found to be heterozygous for at least one MEN1 intragenic polymorphism (25 cases) or for a flanking gene polymorphism (6 cases). The remaining tumours were not informative. No mutations were found in any tumour except in one prolactinoma which was homozygous or hemizygous for a mutation (1-117 C-->T) in a region close to the promoter. Unfortunately, blood or normal tissue was not available in this case. Our data show that somatic MEN1 mutations do not contribute significantly to tumorigenesis of sporadic pituitary adenomas and suggest that mutation of other genes are likely to contribute to the pathogenesis of these tumours. PMID- 10366411 TI - Tissue-specific pattern of variant transcripts of the human gonadotropin releasing hormone receptor gene. AB - The expression pattern of the GnRH receptor was investigated in a variety of normal and neoplastic human tissues by RT-PCR-Southern blotting. In addition to the full-length cDNA (sb1), we identified two other transcripts: the first (sb2) was characterized by a 128 bp deletion as previously described; the second was an unexpected finding composed of a shorter cDNA (sb3), the sequence of which revealed a 220 bp deletion corresponding in size to exon 2. These three transcripts were found in normal pituitary and pituitary adenomas, and in granulosa tumors, but not in testis, where sb2 was lacking. Only sb1 was expressed in normal, fibrocystic and malignant breast tissue. No transcript with a full-length region was found in endometrium, intestine or lymphocytes. This is the first report that shows that splicing of the gonadotropin-releasing hormone receptor gene is tissue dependent. We also determined the intron-exon nucleotide sequence of the gene and identified an MaeIII polymorphic site in exon 1 created by a silent C453T transition found in 10% of unrelated French whites. PMID- 10366413 TI - IGF-I production by adult rat hepatocytes is stimulated by transforming growth factor-alpha and transforming growth factor-beta1. AB - Previously, we have observed that epidermal growth factor (EGF), a potent mitogen for cultured hepatocytes, stimulates the production of IGF-I and IGF-binding proteins (IGFBPs) by cultured hepatocytes from adult rats. This study was undertaken to investigate the possibility that other growth factors of hepatic origin could specifically be involved in the regulation of IGF-I and IGFBP expression. The effects of transforming growth factor-alpha (TGF-alpha), through EGF receptors to induce a mitogenic response, and transforming growth factor beta1 (TGF-beta1), produced by non-parenchymal liver cells and able to inhibit hepatocyte proliferation in vivo and in culture, have been studied in cultured adult rat hepatocytes. Our results demonstrate that TGF-alpha and TGF-beta1 significantly stimulate IGF-I and IGFBP secretion by cultured hepatocytes but no change in the abundance of IGF-I and IGFBP mRNAs was observed with respect to controls. Cycloheximide is able to inhibit both basal and TGF-stimulated release of IGF-I and a similar effect was elicited by octreotide, the somatostatin analog, known to directly affect hepatic IGF-I gene expression. Our findings show the role of the liver in the secretion of IGF-I and IGFBPs, not only under endocrine and nutritional control but also under autocrine and paracrine control. PMID- 10366414 TI - Stimulation of matrix-metalloproteinase-1 and tissue inhibitor of metalloproteinase-1 gene expression in rats by the preovulatory prolactin peak. AB - Since structural luteolysis involves deterioration of tissue, the gene expression of matrix-metalloproteinase-1 (MMP-1) and the respective tissue inhibitor of this metalloprotease (TIMP-1) were measured at various times on the day of pro-oestrus and in animals in which the preovulatory prolactin surge was blocked for the duration of 3 cycles by bromocriptine. An additional group of prolactin-blocked rats received a prolactin replacement injection on the afternoon of pro-oestrus. In spontaneously pro-oestrous rats, MMP-1 and TIMP-1 gene expression increased significantly (P<0.01) prior to the occurrence of the preovulatory LH surge but simultaneously with the onset of the preovulatory prolactin surge. When prolactin release was blocked by bromocriptine for 3 cycles, no such changes were observed during the afternoon of pro-oestrus. However, an intraperitoneal injection of bovine prolactin at the time when the preovulatory prolactin surge occurs normally, increased MMP-1 and TIMP-1 gene expression (P<0.01). These results indicate that MMP-1 and TIMP-1 gene expression are stimulated by the preovulatory prolactin surge. Previous work has shown that the preovulatory LH surge activates the enzymatic cascade which leads to increased collagenase activity. PMID- 10366415 TI - pG2 gene expression and its regulation in human adrenocortical and medullary tumors. AB - The cDNA clone pG2 was originally isolated from a human pheochromocytoma. The respective gene was found to be strongly expressed in normal adrenal zona glomerulosa and medulla, as well as in Conn's adenomas and pheochromocytomas. To shed more light on the expression and regulation of the pG2 gene, we investigated its expression in a wide variety of different adrenal neoplasms and cultured adrenal cells. Northern blot analysis was used to determine the steady state level of pG2 mRNA. Besides normal adrenals, Conn's adenomas and pheochromocytomas, we found abundant expression of pG2 mRNA in Cushing's, virilizing and nonfunctional adrenocortical adenomas and carcinomas, as well as in hyperplastic adrenals. The relative levels of pG2 mRNA in various adrenocortical tumors were not significantly different from those in normal adrenals and pheochromocytomas. In primary cultures of normal adrenal cells, treatment with adrenocorticotropin induced a 3- to 15-fold increase in the expression of pG2 mRNA (P<0.01), and this effect was reproduced by incubation with (Bu)2cAMP. In cultured pheochromocytoma cells, treatment with (Bu)2cAMP and a protein kinase inhibitor, staurosporine, increased pG2 mRNA accumulation (2- to 4-fold over the control level, P<0.01, and 3- to 8-fold, P<0.01, respectively). These results indicate that pG2 is widely expressed in normal and pathological adrenal tissues from both cortical and medullary origin, which eliminates its usefulness as a specific marker for zona glomerulosa or medullary adrenal tumors. Accumulation of pG2 mRNA is regulated by multiple differentiating factors through different pathways in primary cultures of normal adrenal and pheochromocytoma cells. PMID- 10366417 TI - 17beta-estradiol induces protein thiol/disulfide oxidoreductases and protects cultured bovine aortic endothelial cells from oxidative stress. AB - OBJECTIVE: To examine whether or not estrogens induced the expression of protein thiol/disulfide oxidoreductases such as protein disulfide isomerase (PDI), thioredoxin (Trx), Trx reductase, and glutaredoxin (Grx) in vascular endothelial cells. METHODS: The regenerative effects of the protein thiol/disulfide oxidoreductases, PDI, Trx and Grx, on oxidatively damaged proteins were assayed using H2O2-inactivated glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a reporter enzyme. The induction of protein thiol/disulfide oxidoreductases and the accumulation of protein adducts generated by lipid peroxidation were examined by Western blotting in estrogen-treated bovine aortic endothelial cells (BAECs). RESULTS: Reduced PDI, Trx and Grx regenerated the H2O2-inactivated GAPDH in vitro. The levels of these protein disulfide oxidoreductases in BAECs were increased by pretreatment with 0.01-10 micromol/l 17beta-estradiol, the largest increase (about fourfold of the control) being found for PDI. Other sex hormones such as progesterone and testosterone did not affect the contents of these oxidoreductases in BAECs. 4-Hydroxy-2-nonenal (HNE)-protein adducts, which are generated by lipid peroxidation, were accumulated in BAECs exposed to paraquat, whereas the pretreatment of BAECs with 17beta-estradiol suppressed their accumulation. CONCLUSIONS: The estrogen-mediated induction of the protein thiol/disulfide oxidoreductases such as PDI, Trx, Trx reductase and Grx suggested a possible involvement of these oxidoreductases in the antioxidant protection of estrogen observed in the vascular system. PMID- 10366416 TI - Regulation of thymosin beta10 expression by TSH and other mitogenic signals in the thyroid gland and in cultured thyrocytes. AB - OBJECTIVE: To investigate the expression of thymosin beta10 - a small conserved acidic protein involved in the inhibition of actin polymerization - in human and experimental thyroid goiters as well as the regulation exerted by TSH on thymosin beta10 expression in thyroid follicular cells both in vivo and in vitro. DESIGN: To this aim, we have used 5 bioptic specimens from patients affected by thyroid goiter, a well known experimental model of thyroid goitrogenesis (rat fed with the drug propylthiouracil) and a cultured rat thyroid cell line (PC Cl 3 cells) as a model system. RESULTS: We report that the mRNA expression of thymosin beta10 is markedly enhanced in human goiters compared with normal thyroid. In vivo results showed that the steady-state level of thymosin beta10 mRNA is up regulated in the thyroid gland of propylthiouracil-fed rats in parallel with follicular cell proliferation: iodide administration to goitrous rats, which induced a marked involution of thyroid hyperplasia, reduced the mRNA level of thymosin beta10. Finally, in vitro studies showed that in cultured rat thyrocytes, the expression of thymosin beta10 mRNA is induced in a time- and dose dependent manner by the activation of pathways which are mitogenic for thyroid cells (i.e. the protein kinase (PK) A and PKC pathways). CONCLUSION: Taken together, the findings reported here demonstrate that thymosin beta10 expression is regulated by extracellular signals that stimulate growth of thyroid cells both in vitro and in vivo, and suggest a role for this protein in thyroid diseases characterized by proliferation of follicular cells. PMID- 10366418 TI - The Sm core domain mediates targeting of U1 snRNP to subnuclear compartments involved in transcription and splicing. AB - In the mammalian cell nucleus pre-mRNA splicing factors such as U snRNPs are concentrated in distinct subnuclear compartments named perichromatin fibrils (PFs), interchromatin granules (IGs), interchromatin granule-associated zones (IG associated zones), and coiled bodies (CBs). The structural requirement for the localization of U snRNPs to these domains was investigated by microinjection of digoxygenin-labeled in vitro-reconstituted U1 snRNPs and mutants thereof and subsequent analysis by immunoelectron microscopy. Wild-type U1 snRNP was targeted, after injection into the cytoplasm, to the nucleus and localized in PFs, IGs, IG-associated zones, and CBs. Thus, microinjected U1 snRNP particles exhibited a subnuclear localization similar to that previously observed for endogenous U1 snRNPs. Specific U snRNP proteins were shown not to be essential for subnuclear targeting since U1 snRNP mutants that did not bind to 70K, A, or C peptides were distributed in the cell nucleus in a pattern indistinguishable from that of wild-type U1 snRNP. Moreover, the Sm core domain, common to all spliceosomal U snRNPs, was shown to be sufficient for appropriate subnuclear distribution. Thus, these observations indicate that the Sm core domain, previously shown to be essential for nuclear import of spliceosomal U1 snRNPs, is also important for mediating the targeting to distinct nuclear subcompartments. PMID- 10366419 TI - Human germ cell tumor cell lines express novel leukemia inhibitory factor transcripts encoding differentially localized proteins. AB - The polyfunctional cytokine leukemia inhibitory factor (LIF) has been implicated in the maintenance of many stem and progenitor cell populations and as an autocrine growth factor for many tumor cell populations, including germ cell tumors. Studies of LIF transcript expression in germ cell tumor cell lines identified two novel human LIF transcripts, hLIF-M and hLIF-T, containing noncoding alternate first exons that are conserved among all reported LIF genes. Embryonal carcinoma (EC) cell lines expressed these transcripts at consistent levels and hLIF-M was generally the predominant LIF transcript in these cells. This expression pattern was characteristic of EC cells since variable independently regulated expression of these transcripts was evident in other cell lines. Overexpression analysis demonstrated that each alternate hLIF transcript generated different levels of extracellular LIF activity as a consequence of the translation of distinct but partially overlapping sets of proteins. Secreted LIF proteins translated from alternate initiation codons were expressed from the hLIF D and hLIF-M transcripts. Intracellular, potentially cell-autonomous, proteins were encoded by the hLIF-M and hLIF-T transcripts. Since EC cell lines also expressed LIF receptor transcripts, the novel LIF transcription profiles and proteins identified here suggest a role for autocrine and/or cell-autonomous LIF signaling during germ cell tumorigenesis. PMID- 10366420 TI - Specific activation of a STAT family member in Xiphophorus melanoma cells. AB - Melanoma formation in the teleost fish Xiphophorus is caused by the uncontrolled activity of the genetically defined tumor locus Tu. The critical component of this locus is the Xmrk oncogene encoding a subclass I receptor tyrosine kinase. Overexpression and constitutive activation of the Xmrk receptor triggers a set of specific signal transduction events eventually resulting in the malignant phenotype. We have identified a melanoma-specific DNA-protein complex which seems to depend on Xmrk activation as shown in a heterologous cell system. The critical component of this complex, which directs transcriptional activation in the melanoma cells, proved to be a fish homologue of STAT5. Two other STAT factors, STAT1 and STAT3, implied in signaling by the Xmrk-related EGF receptor, were not activated in this particular cell type. Thus, Xmrk initiates very specific signaling pathways and transcriptional responses in Xiphophorus melanoma. PMID- 10366421 TI - Subcellular localization of RhoA and ezrin at membrane ruffles of human endothelial cells: differential role of collagen and fibronectin. AB - Endothelial cells and the regulation of their migration are of prime importance in many physiological and pathological processes such as angiogenesis. RhoA, an important Rho family member known to trigger actin reorganization, has been shown to mediate the formation of focal adhesions and stress fibers in quiescent fibroblasts. However, recent studies have emphasized its functional diversity and its implication in migration or metastatic processes in different cell types other than fibroblasts. Its role in endothelial cells is little known. In this study, we were interested by analyzing in human endothelial cells the subcellular redistribution of endogenous RhoA and the reorganization of cytoskeletal actin induced by two important extracellular matrix proteins, collagen and fibronectin. This paper shows a translocation of RhoA and its association with cortical actin in focal contact domains at membrane ruffles and at lamellipodia of spread or migrating endothelial cells, in the absence of any soluble mitogen stimulation. Furthermore, RhoA was found colocalized with ezrin, a member of the ERM family proteins newly described as important membrane-actin cytoskeleton linkers, at early membrane ruffles of endothelial cells spread on collagen but not on fibronectin. The present study points out that extracellular matrix, depending on the nature of its components, may promote distinct assemblies of focal contact constitutive proteins and strongly suggests that endothelial RhoA, like Rac1, may be an important mediator of matrix signaling pathway regulating endothelial cell adhesiveness and motility, independently of growth factor stimulation. PMID- 10366422 TI - Alpha 2 beta 1 integrin mediates dermal fibroblast attachment to type VII collagen via a 158-amino-acid segment of the NC1 domain. AB - Dermal fibroblasts are in apposition to type VII (anchoring fibril) collagen in both unwounded and wounded skin. The NC1 domain of type VII collagen contains multiple submodules with homology to known adhesive molecules, including fibronectin type III-like repeats and a potential RGD cell attachment site. We previously reported the structure and matrix binding properties of authentic and recombinant NC1. In this study, we examined the interaction between dermal fibroblasts and the NC1 domain of type VII collagen. We found that both recombinant and authentic NC1 vigorously promoted human fibroblast attachment. Adhesion of fibroblasts to NC1 was dose dependent, saturable, and abolished by both polyclonal and monoclonal antibodies to NC1. Cell adhesion to NC1 was divalent cation dependent and specifically inhibited by a monoclonal antibody directed against the alpha2 or beta1 integrin subunits, but not by the presence of RGD peptides. Furthermore, the cell-binding activity of NC1 was not conformation dependent, since heat-denatured NC1 still promoted cell adhesion. Using a series of recombinant NC1 deletion mutant proteins, the cell binding site of NC1 was mapped to a 158-aa (residues 202-360) subdomain. We conclude that human dermal fibroblasts interact with the NC1 domain of type VII collagen and this cell-matrix interaction is mediated by the alpha2beta1 integrin and is RGD independent. PMID- 10366423 TI - NA22598, a novel antitumor compound, reduces cyclin D1 levels, arrests cell cycle at G1 phase, and inhibits anchorage-independent growth of human tumor cells. AB - NA22598, a novel antitumor compound isolated from a microbial cultured broth, inhibited the growth of human colon cancer DLD-1 cells in suspension cultures (anchorage-independent growth) severalfold more strongly than in substratum attached monolayer cultures. It arrested the cell cycle progression at early G1 phase under both these culture conditions. Rb phosphorylation, cyclin D1 expression, and cdk2 activation in G1 progression were all inhibited by NA22598, but the amounts of cdk2 and p27 were not affected. Among these effects the inhibition of cyclin D1 expression was most prominent, and NA22598 was found to inhibit the synthesis of cyclin D1 without affecting mRNA expression or protein degradation. p27 binding to cdk2 was more markedly increased in suspension cultures than in attached cultures by NA22598, but the compound had no effect on total p27. Apparently, the decrease of cyclin D1 induced redistribution of p27 from the cyclin D1/cdk4 to the cyclin E/cdk2 complexes during G1 phase in the suspension cultures. Because p27 is upregulated during suspension culture, a greater amount of it was associated with cyclin E/cdk2, thus producing greater growth inhibition. An agent, like NA22598, which induces the downregulation of cyclin D1 might offer a new anticancer strategy. PMID- 10366424 TI - SV40 large T antigen expression driven by col2a1 regulatory sequences immortalizes articular chondrocytes but does not allow stabilization of type II collagen expression. AB - Immortalization of chondrocytes by SV40 T Ag has often been reported to trigger the loss of expression of type II collagen, one of the main differentiation markers, although some immortalized chondrocyte lines maintaining a differentiated phenotype have also been described. Here, we show using transient cotransfections in differentiated chondrocytes that, in contrast to c-src, neither SV40 T Ag, nor c-myc, decreases col2a1 transcriptional activity. Then, we report the possibility of immortalizing rabbit articular chondrocytes by expression of SV40 T Ag controlled by the col2a1 promoter and enhancer (pCol2SV). This strategy allows one to select within a population of differentiated chondrocytes those which are able to maintain functional regulation of the col2a1 gene through long-term culture. In precrisis pCol2SV-transfected chondrocytes, all-trans-retinoic acid, a down-regulator of col2a1 expression, induced apoptosis, strongly suggesting the strict control of T Ag expression by col2a1 regulatory sequences. Some pCol2SV-transfected chondrocytes were definitively immortalized, after a short crisis period. However, type II collagen synthesis was restricted to a small proportion of cells, which went on to decrease with subculture, while the proportion of cells expressing T Ag was not affected. In these postcrisis cells, T Ag remained at least partially under the control of functional col2a1 regulatory elements as assessed by all-trans-retinoic acid down regulation. PMID- 10366425 TI - Protease activation in apoptosis induced by MAL. AB - The proteolytic caspase cascade plays a central role in the signaling and execution steps of apoptosis. This study investigated the activation of different caspases in apoptosis induced by MAL (a folding variant of human alpha lactalbumin) isolated from human milk. Our results show that the caspase-3-like enzymes, and to a lesser extent the caspase-6-like enzymes, were activated in Jurkat and A549 cells exposed to MAL. Activated caspases subsequently cleaved several protein substrates, including PARP, lamin B, and alpha-fodrin. A broad range caspase inhibitor, zVAD-fmk, blocked the caspase activation, the cleavage of proteins, and DNA fragmentation, indicating an important role for caspase activation in MAL-induced apoptosis. Since an antagonistic anti-CD95 receptor antibody, ZB4, did not influence the MAL-induced killing, we conclude that this process does not involve the CD95-mediated pathway. While MAL did not directly activate caspases in the cytosol, it colocalized with mitochondria and induced the release of cytochrome c. Thus, these results demonstrate that caspases are activated and involved in apoptosis induced by MAL and that direct interaction of MAL with mitochondria leads to the release of cytochrome c, suggesting that this release is an important step in the initiation and/or amplification of the caspase cascade in these cells. PMID- 10366426 TI - Endogenous interleukin 6 conveys resistance to cis-diamminedichloroplatinum mediated apoptosis of the K562 human leukemic cell line. AB - Cisplatin is an effective chemotherapeutic agent that elicits its antineoplastic activity by binding to DNA and disrupting template functions. IL-6 is a cytokine which has been shown to play a central role in host immunological defense mechanisms. Although K562 leukemic cells have been shown to secrete IL-6, little is known of whether there exists a correlation between the expression of IL-6 and the resistance of these cells to anticancer chemotherapeutic agents. To determine the contribution of IL-6 to the regulation of cisplatin-induced apoptosis in K562 cells, we examined whether treatment of K562 cells and cisplatin-resistant K562 subclones with anti-IL-6 mAb enhances their sensitivity to cisplatin. The results show that cis-diamminedichloroplatinum (CDDP) resistance was overcome by treatment with nontoxic doses of CDDP in combination with anti-IL-6 mAb. When we tested if the synergistic effect of anti-IL-6 and cisplatin could restore the ability of K562 mutant cells to undergo apoptosis, we found the typical DNA laddering in these cells, even in the presence of a nontoxic dose of the drug. Treatment of cells with anti-IL-6 reduced the levels of glutathione. The current studies show that anti-IL-6 mAb sensitized CDDP-resistant K562 cells to CDDP by induction of apoptotic death and the reduction of glutathione levels might be implicated in the enhanced cytotoxicity observed. PMID- 10366427 TI - Role of vitamin D3 receptor in the synergistic differentiation of WEHI-3B leukemia cells by vitamin D3 and retinoic acid. AB - WEHI-3B D- cells differentiate in response to 1,25-dihydroxyvitamin D3 (1,25 (OH)2D3) but not to all-trans-retinoic acid (RA) or other inducing agents. Combinations of RA with 1,25-(OH)2D3 interact to produce synergistic differentiation of WEHI-3B D- cells. To determine factors involved in the synergistic interaction, expression of the 1,25-(OH)2D3 receptor (VDR) and retinoid receptors, RARalpha and RXRalpha, was measured. No VDR was detected in untreated WEHI-3B D- cells; however, RA and 1,25-(OH)2D3 when used as single agents caused a slight induction of the VDR and in combination produced a marked increase in the VDR. In contrast, no changes in RARalpha and RXRalpha were initiated by these compounds. An RAR-selective agonist combined with 1,25-(OH)2D3 produced synergistic differentiation of WEHI-3B D- cells, whereas an RXR selective agonist did not. To gain information on the role of the VDR in the synergistic interaction, the VDR gene was transferred into WEHI-3B D+ cells, in which no VDR was detected and no synergism was produced. Expression of the VDR conferred differentiation responsiveness to 1,25-(OH)2D3 in WEHI-3B D+ cells. These findings suggest that (a) induction of VDR expression is a key component in the synergistic differentiation induced by 1,25-(OH)2D3 and RA and (b) RAR and not RXR must be activated for enhanced induction of the VDR and for the synergistic differentiation produced by RA and 1, 25-(OH)2D3. PMID- 10366428 TI - Fas antigen modulates ultraviolet B-induced apoptosis of SVHK cells: sequential activation of caspases 8, 3, and 1 in the apoptotic process. AB - Interferon-gamma (IFN-gamma) induces various apoptosis-related proteins, including Fas antigen (Fas) in keratinocytes. Ultraviolet B (UVB) irradiation produces "sunburn cells," a specific type of apoptosis. Previously, we reported that IFN-gamma augments Fas-dependent apoptosis of SV40-transformed human keratinocytes (SVHK cells). Caspases are a new class of cysteine proteinases that play an important role in apoptosis. We investigated the mechanism of UVB-induced apoptosis by examining activation of the caspase cascade. UVB irradiation of SVHK cells increased the activities of caspases 1, 3, and 8, which were detected at 3 h, and peak activities occurred at 6 h. Pretreatment of SVHK cells with IFN-gamma significantly increased the activity of caspases 1, 3, and 8. UVB-induced caspase 8 stimulation was significantly suppressed only by caspase 8 inhibitor, while inhibitors of caspases 1, 3, and 8 significantly suppressed UVB-induced caspase 1 stimulation. Caspase 3 and 8 inhibitors, but not caspase 1 inhibitor, significantly suppressed UVB-induced caspase 3 activity, suggesting sequential activation of caspases 8, 3, and 1 in UVB-irradiated SVHK cells. Cross-linking and immunoprecipitation analyses showed multimerization of Fas antigen following UVB irradiation of SVHK cells. Pretreatment of SVHK cells with IFN-gamma significantly augmented UVB-induced apoptosis that was accompanied by increased Fas expression. The susceptibility to UVB-induced apoptosis was also increased in Fas-transfected SVHK cells (F2 cells). Neutralizing anti-Fas antibody significantly suppressed caspase activation and Fas-dependent apoptosis of SVHK cells and F2 cells. In contrast, UVB-induced caspase activation and apoptosis were not inhibited by neutralizing anti-Fas antibody in both cell lines. Our results suggest that UVB directly activates Fas and subsequent caspase cascade resulting in apoptosis of SVHK cells. Furthermore, the expression level of Fas antigen in keratinocytes influenced their susceptibility to UVB-induced apoptosis. PMID- 10366429 TI - A novel apoptosis-like pathway, independent of mitochondria and caspases, induced by curcumin in human lymphoblastoid T (Jurkat) cells. AB - We have shown previously [E. Sikora, A. Bielak-Zmijewska, K. Piwocka, J. Skierski, and E. Radziszewska (1997) Biochem. Pharmacol. 54, 899-907] that curcumin prevents formation of oligonucleosomal DNA fragmentation in rat thymocytes and human leukemic T lymphocytes (Jurkat cells) induced to undergo apoptosis. In this paper we show that 50 microM curcumin by itself induces cell death in Jurkat cells, but its symptoms differ from those observed after a short ultraviolet (uv) irradiation. Ultraviolet-irradiated Jurkat cells displayed typical symptoms of apoptosis: morphological changes, internucleosomal and high molecular-weight DNA fragmentation, formation of sub-G1 fractions in DNA content frequency histograms, and dissipation of the mitochondrial transmembrane electric potential (Delta psi). In contrast, curcumin-treated Jurkat cells exhibited DNA splitting into high-, but not low-, molecular-weight fragments. These cells retained their high mitochondrial Delta psi, and the content of Ca2+ in endoplasmic reticulum stores remained at the level typical for untreated cells. The frequency of opening of the mitochondrial permeability transition pores in curcumin-treated cells was decreased compared to the controls, whereas uv irradiation made these pores completely open. Curcumin did not produce any change in the activity of caspase-3, whereas uv irradiation considerably activated this protease. The morphology of curcumin-treated cells displayed chromatin condensation, which was insensitive to the caspase inhibitor z-VAD-fmk, but no formation of typical apoptotic bodies, as was the case after uv irradiation. In contrast to uv-irradiated cells, curcumin-treated Jurkat cells considerably increased the level of Bcl-2. It is concluded that the programmed cell death induced by curcumin in Jurkat cells differs from "classical" by the lack of mitochondrial depolarization and of the involvement of caspases. PMID- 10366430 TI - The cytochrome b5 tail anchors and stabilizes subdomains of human DNA topoisomerase II alpha in the cytoplasm of retrovirally infected mammalian cells. AB - DNA topoisomerase II (topo II) is the target of many anticancer drugs and is often altered in drug-resistant cell lines. In some tumor cell lines truncated isoforms of topo IIalpha are localized to the cytoplasm. To study the localization and function of individual enzyme domains, we have epitope-tagged several fragments of human topo IIalpha and expressed them by retroviral infection of rodent and human cells. We find that fusion of the topo II fragments to the hydrophobic tail of human liver cytochrome b5 anchors the fusion protein to the outer face of cytoplasmic membranes, as determined by colocalization with calnexin and selective detergent permeabilization. Moreover, whereas the minimal ATPase domain (aa 1-266) is weakly and diffusely expressed, addition of the cytb5 anchor (1-266-b5) increases its steady-state level 16-fold with no apparent toxicity. Similar results are obtained with the complete ATPase domain (aa 1 426). A C-terminal domain (aa 1030-1504) of human topo IIalpha containing an intact dimerization motif is stably expressed and accumulates in the nucleus. Fusion to the cytb5 anchor counteracts the nuclear localization signal and relocalizes the protein to cytoplasmic membranes. In conclusion, we describe a technique that stabilizes and targets retrovirally expressed proteins such that they are exposed on the cytoplasmic surface of cellular membranes. This approach may be of general use for regulating the nuclear accumulation of drugs or proteins in living cells. PMID- 10366431 TI - Tex27, a gene containing a zinc-finger domain, is up-regulated during the haploid stages of spermatogenesis. AB - Tex27 is a gene encoding a protein containing a zinc-finger domain in the carboxy terminal region and a transactivation domain in the amino terminal region. The Tex27 cDNA was isolated from a subtractive library that was enriched for genes preferentially expressed during the development of the seminiferous epithelium. Northern and in situ hybridization analyses demonstrated that Tex27 is differentially expressed in the testis, showing an increased expression in the germ cells corresponding to postmeiotic stages of spermatogenesis. This expression pattern in testis has been described for other C2H2-type zinc-finger proteins in mouse and human, like CTfin51, Zpf29, Sp1, and Zpf37. RFLP-Southern assays revealed that Tex27 is conserved in mammals. The polypeptide analysis and expression pattern suggest that Tex27 is a potential transcription factor preferentially expressed in postmeiotic cells during mouse spermatogenesis. PMID- 10366432 TI - Dissociation of CDK2 from cyclin A in response to the topoisomerase II inhibitor etoposide in v-src-transformed but not normal NIH 3T3 cells. AB - Our previous work has demonstrated that treatment of NIH 3T3 cells with etoposide (VP16), an inhibitor of DNA topoisomerase II and widely used anticancer agent, results in G2/M-phase arrest, whereas treatment of cells transformed by v-src, v ras, or v-raf results in an S-phase blockage. The present studies describe the mechanistic aspects of this selective S-phase arrest in the v-src-transformed cells. The S-phase arrest in these cells was found to be coupled with depletion of cyclin A-dependent kinase activity. This decrease could not be explained by changes in the overall level of cyclin A, CDK2, p27, or p21 proteins. Rather, it was associated with a time-dependent reduction of CDK2 protein complexed with cyclin A following VP16 treatment. It was further shown that the decrease of cyclin A-associated CDK2 was linked to an increase of CDK2 protein in cyclin E immunocomplexes, which suggests that CDK2 might become redistributed following treatment with VP16. Thus, oncogenic transformation by v-src can trigger separation of CDK2 protein from cyclin A in response to VP16. This might contribute to the depletion of cyclin A-dependent kinase activity and the selective S-phase arrest by VP16 in v-src-transformed cells. PMID- 10366433 TI - BTG1: a triiodothyronine target involved in the myogenic influence of the hormone. AB - The product of the B-cell translocation gene 1 (BTG1), a member of an antiproliferative protein family including Tis-21/PC3 and Tob, is thought to play an important role in the regulation of cell cycle progression. We have shown in a previous work that triiodothyronine (T3) stimulates quail myoblast differentiation, partly through a cAMP-dependent mechanism involved in the stimulation of cell cycle withdrawal. Furthermore, we found that T3 or 8-Br-cAMP increases BTG1 nuclear accumulation in confluent myoblast cultures. In this study, we report that BTG1 is essentially expressed at cell confluence and in differentiated myotubes. Whereas neither T3 nor cAMP exerted a direct transcriptional control upon BTG1 expression, we found that AP-1 activity, a crucial target involved in the triiodothyronine myogenic influence, repressed BTG1 expression, thus probably explaining the low BTG1 expression level in proliferating myoblasts. In transient transfection studies, we demonstrated that an AP-1-like sequence located in the BTG1 promoter was involved in this negative regulation. Our present data also bring evidence that the stimulation of BTG1 nuclear accumulation by T3 or 8-Br-cAMP probably results from an increased nuclear import or retention in the nucleus. Lastly, BTG1 overexpression in quail myoblasts mimicked the T3 or 8-Br-cAMP myogenic influence: (i) inhibition of myoblast proliferation due to an increased rate of myoblast withdrawal from the cell cycle; and (ii) stimulation of terminal differentiation. These data suggest that BTG1 is probably involved in T3 and cAMP myogenic influences. In conclusion, BTG1 is a T3 target involved in the regulation of myoblast differentiation. PMID- 10366435 TI - Maintenance of functional stem cells in isolated and cultured adult intestinal epithelium. AB - We have previously described a method for the primary culture of adult large intestinal epithelium, suggesting that stem cells had survived both the isolation and the culture procedures. However, as no markers for such cells exist, confirmation of stem cell survival is difficult-only the functional properties can be used to define them. Unfortunately, many of these (e.g., differentiation, crypt regeneration) do not occur in culture, probably due to suboptimal conditions. To address this problem both freshly isolated and cultured small and large intestinal crypts were grown subcutaneously in an immunocompromized mouse. All initially formed cysts lined by a simple epithelium which gradually became multicellular and formed invaginations containing many mitoses and apoptoses. Epithelial differentiation, as assayed by Goblet cell mucin production, was also apparent. Mucin maturation was also typical of the normal intestine. The lumen was frequently filled with mucin and apoptotic bodies. Interestingly, in grafts displaying pronounced crypt-like morphology the regions of proliferation were situated toward the base of the structure and the Goblet cells toward the lumen, i.e., a typical crypt-like morphology. Hence, functional adult stem cells appear to survive isolation and tissue culture, permitting organotypic regeneration, possibly involving homeobox gene expression. This may now allow direct stem cell characterization and experimental manipulation, such as transfection, and may ultimately permit transplantation and therapeutic gene therapy. PMID- 10366434 TI - Conversion of TNF alpha from antiproliferative to proliferative ligand in mouse intestinal epithelial cells by regulating mitogen-activated protein kinase. AB - The mechanisms regulating the balance between intestinal epithelial cell proliferation and differentiation are essential to maintaining an intact mucosal barrier. Mitogen-activated protein (MAP) kinases appear to be key transducers of extracellular signals in these pathways. The goal of this study was to investigate the regulation of MAP kinase by tumor necrosis factor alpha (TNFalpha) and epidermal growth factor (EGF) in intestinal epithelial cells. The young adult mouse colon cell line was studied for TNFalpha and/or EGF regulation of MAP kinase in the presence or absence of the MAP kinase kinase (MEK1) inhibitor PD 98059. Proliferation was determined by hemocytometry, and activated MAP kinase was identified by Western blot analysis, in vitro kinase assay, and confocal laser immunofluorescent microscopy. TNFalpha stimulated sustained nuclear MAP kinase activity, while EGF stimulated transient cytoplasmic MAP kinase activity. Changing TNFalpha's sustained MAP kinase activation to transient converted TNFalpha from an anti-proliferative to a proliferative ligand. These findings demonstrate that both TNFalpha and EGF activate MAP kinase in intestinal epithelial cells. The kinetics and subcellular distribution of this enzyme activity may be pivotal in the transduction of divergent cellular responses in the intestinal epithelium with implications for altered proliferative signals in inflammatory bowel disease. PMID- 10366436 TI - Expression of a Cx43 deletion mutant in 3T3 A31 fibroblasts prevents PDGF-induced inhibition of cell communication and suppresses cell growth. AB - Communication through gap junctions was first suggested to have a role in the social control of cell growth over 30 years ago. However, despite extensive experimentation, the importance of gap junctions as a general mechanism of growth control remains to be established. A number of different studies have shown that a common early response of cells in culture to polypeptide growth factors such as PDGF is a rapid and transient inhibition of cell communication suggesting that a cell may have to lose communication with its neighbors before it can undergo cell division. Here we show that 3T3 A31 fibroblasts exposed to PDGF exhibit a 50% decrease in cell communication as measured by dye transfer in the absence of significant changes in the cellular content and distribution of Cx43. Likewise, PDGF inhibited cell communication in cells transfected either with a vector which did not contain a cDNA or with an expression vector encoding full-length Cx43 fused to a c-myc tag (Cx43-M). In contrast, 3T3 A31 fibroblasts transfected with an expression construct encoding a deletion mutant of Cx43 (Cx43-256M) consisting of amino acids 1-256 of Cx43 fused to a c-myc tag maintain high levels of gap junction activity following exposure to PDGF. These results suggest that sites which trigger loss of cell communication in response to PDGF are located within amino acids 257 to 382 of the Cx43 molecule. Cells transfected with an expression vector encoding full-length Cx43 fused to a c-myc tail exhibited a reduced basal growth rate compared to both parent cells and cells transfected with a control vector but maintained a strong mitogenic response to PDGF. In contrast, both the basal growth rate and the mitogenic response to PDGF was markedly reduced in cells which expressed Cx43-256M consistent with the hypothesis that loss of cell communication is required before a cell can respond to mitogenic stimuli. PMID- 10366437 TI - Terminal differentiation of normal human oral keratinocytes is associated with enhanced cellular TGF-beta and phospholipase C-gamma 1 levels and apoptotic cell death. AB - Subculture of primary normal human oral keratinocytes (NHOK) results in terminal differentiation, leading to cell death. To investigate whether the subculture induced death of NHOK is due to apoptosis, we studied transferase-mediated dUTP nick end labeling (TUNEL)-positive cells, DNA fragmentation, and expression of several apoptosis-associated genes from NHOK with different passage numbers. We also determined the effect of transforming growth factor beta1 (TGF-beta1) on the induction of apoptosis in NHOK. We were able to subculture primary NHOK up to the fifth passage, at which point cells showed morphological features of differentiation. Appearance of DNA fragmentation concurrently occurred with an increase in the number of TUNEL-positive cells with higher passage numbers. The level of cellular p53 proteins was gradually decreased by the continued passage of cells, whereas the levels of intracellular and secreted TGF-beta and phospholipase C-gamma1 (PLC-gamma1) were significantly elevated by serial subculture. Exogenous TGF-beta1 also induced differentiation and apoptosis of proliferating NHOK. These data indicate that terminal differentiation of NHOK is associated with apoptosis, which is, in part, linked to elevated cellular levels of TGF-beta and PLC-gamma1. PMID- 10366438 TI - Identification of metastasis-promoting sequences in the mouse laminin alpha 1 chain. AB - Laminin-1, a major basement membrane matrix glycoprotein, enhances adhesion, migration, and metastasis of tumor cells. We have screened 208 overlapping synthetic peptides covering the short and long arms of mouse laminin alpha1 chain for their adhesion activity with B16-F10 mouse melanoma cells. Cell adhesion activity was determined using various amounts of peptides coated on plastic dishes and by measuring cell adhesion on peptide-conjugated Sepharose beads. Nineteen peptides showed B16-F10 cell adhesion activity. Three peptides, designated A-13, -24, and -208, showed the strongest attachment activity in the plate assay, whereas 4 peptides, A-13, -51, -99, and -112, demonstrated the strongest cell adhesion when conjugated to beads. The 19 peptides were tested in vivo for their effect on experimental pulmonary metastasis by B16-F10 cells. Four peptides, A-13, -51, -64, and -119, significantly enhanced metastasis, with A-13 showing the strongest dramatic enhancement. The four metastasis-promoting peptides also stimulated migration of B16-F10 cells in the Boyden chamber assay in vitro with A-13 being the most potent stimulator. In addition, the 4 peptides inhibited laminin-induced cell attachment and migration, which indicates that these four sequences are possible functional B16-F10 cell binding sites in laminin-1. All the four sequences are located on the globular domains of the short arm. Other peptides, including strong adhesion-active peptides, A-24, -99, 112, and a scrambled A-13 peptide, did not stimulate either migration or metastasis. Thus, laminin-1 has multiple active sites in the globular domains of the short arm which promote migration and metastasis of B16-F10 cells. PMID- 10366439 TI - Inhibition of mitochondrial ATP generation by nitric oxide switches apoptosis to necrosis. AB - Under pathological conditions, the mode of cell death, apoptosis or necrosis, is relevant for the subsequent fate of the tissue. Cell demise may be shaped by endogenous mediators such as nitric oxide (NO) which interfere with subroutines of the death program. Here we show that apoptosis of Jurkat cells elicited by either staurosporine (STS) or anti-CD95 antibodies in glucose-free medium is converted to necrosis by NO donors. In the presence of NO, release of mitochondrial cytochrome c was delayed and activation of execution caspases was prevented. Stimulated cells died nonetheless. The switch in the mode of cell death was due to NO-dependent failure of mitochondrial energy production. Restoration of intracellular ATP by glucose supplementation recovered the cells' ability to activate caspases and undergo apoptosis. In this system, the apoptosis/necrosis conversion promoted by NO was not mediated by cyclic guanosine monophosphate-dependent mechanisms, poly-(ADP-ribose)-polymerase (PARP) activation, or inhibition of caspases due to S-nitrosylation and glutathione depletion. In contrast, depleting intracellular ATP with rotenone, an inhibitor of mitochondrial complex I mimicked the effect of NO. The findings presented here suggest that NO can decide the shape of cell death by lowering intracellular ATP below the level required to allow the coordinated execution of apoptosis. PMID- 10366440 TI - The Schrodinger's cat quandary in cell biology: integration of live cell functional assays with measurements of fixed cells in analysis of apoptosis. AB - The existing cytometric methodologies do not allow one to directly correlate, within the same cells, functional cell attributes that are revealed supravitally with features that require cell fixation to be detected or measured. Taking advantage of the "file merge" feature of the laser-scanning cytometer, we have been able to correlate the supravital changes that occur during apoptosis, namely the drop in mitochondrial transmembrane potential (Delta Psim) and generation of the reactive oxygen intermediates (ROIs), with features revealed by analysis of fixed cells: the cell cycle position and DNA fragmentation. The cell cycle position was established based on the cell's stainability with propidium iodide while DNA fragmentation was assessed by in situ DNA strand break labeling using exogenous terminal deoxynucleotidyltransferase. During apoptosis of HL-60 cells induced by the DNA topoisomerase I inhibitor camptothecin (CPT), the dissipation of Delta Psim occurred preferentially in S-phase cells and preceded the appearance of DNA strand breaks. Essentially all cells with DNA strand breaks had dissipated Delta Psim. Compared to the decrease of Delta Psim, the CPT-induced rise in ROIs during apoptosis was less restricted to S-phase cells. Furthermore, no elevation of ROIs was detected in a significant proportion of cells with DNA strand breaks. The data suggest that DNA fragmentation may occur in some cells prior to the increase in ROIs and thus, unlike the dissipation of Delta Psim, the oxidative stress may not be a prerequisite for activation of an endonuclease. Alternatively, the oxidative stress may be a transient event, occupying a narrow "time window" during the apoptotic process. The approach opens a possibility to study direct relationships, within the same cells, between cellular changes (e.g., occurring during apoptosis, mitogenesis, differentiation, etc.) detected by functional assays of live cells and changes that cannot be analyzed supravitally. PMID- 10366442 TI - Cyclin-dependent kinase 5 activity enhances monocytic phenotype and cell cycle traverse in 1,25-dihydroxyvitamin D3-treated HL60 cells. AB - The function of most cyclin-dependent kinases (Cdks) is to facilitate progression through the checkpoints of the cell cycle, but Cdk5 is known to be involved in differentiation of CNS, muscle, and lens cells, though not in the cell cycle traverse. Here we show an additional role for Cdk5, an enhancement of monocytic differentiation with abrogation of the G1 checkpoint. Human leukemia HL60 cells exposed to 1alpha,25-dihydroxyvitamin D3 (1,25D3) displayed monocytic phenotype and increased Cdk5 kinase activity. An analog of 1,25D3 which does not induce differentiation failed to upregulate Cdk5, and 1,25D3-resistant cells had reduced Cdk5 activity. Active or inactive Cdk5 was associated with cyclin D1, but only active Cdk5 exhibited threonine phosphorylation. Inhibition of Cdk5 expression by an antisense construct reduced the intensity of 1, 25D3-induced expression of CD14, a marker of monocytes, and increased the 1,25D3-induced G1 block. These findings demonstrate a novel aspect of Cdk5 activity-facilitation of the G1- to S phase transition in cells which are approaching replicative quiescence and a concomitant enhancement of monocytic differentiation. PMID- 10366441 TI - Arsenite induces apoptosis via a direct effect on the mitochondrial permeability transition pore. AB - The molecular mode of action of arsenic, a therapeutic agent employed in the treatment of acute promyelocytic leukemia, has been elusive. Here we provide evidence that arsenic compounds may act on mitochondria to induce apoptosis. Arsenite induces apoptosis accompanied by a loss of the mitochondrial transmembrane potential (Delta Psim). Inhibition of caspases prevents the arsenite-induced nuclear DNA loss, but has no effect on the Delta Psim dissipation and cytolysis induced by arsenite. In contrast, Bcl-2 expression induced by gene transfer prevents all hallmarks of arsenite-induced cell death, including the Delta Psim collapse. PK11195, a ligand of the mitochondrial benzodiazepine receptor, neutralizes this Bcl-2 effect. Mitochondria are required in a cell-free system to mediate arsenite-induced nuclear apoptosis. Arsenite causes the release of an apoptosis-inducing factor (AIF) from the mitochondrial intermembrane space. This effect is prevented by the permeability transition (PT) pore inhibitor cyclosporin A, as well as by Bcl-2, which is known to function as an endogenous PT pore antagonist. Arsenite also opens the purified, reconstituted PT pore in vitro in a cyclosporin A- and Bcl-2-inhibitible fashion. Altogether these data suggest that arsenite can induce apoptosis via a direct effect on the mitochondrial PT pore. PMID- 10366443 TI - Molecular basis of feline beta-glucuronidase deficiency: an animal model of mucopolysaccharidosis VII. AB - A family of domestic cats was found that exhibited clinical and biochemical abnormalities consistent with mucopolysaccharidosis VII, an autosomal recessive lysosomal storage disorder caused by beta-glucuronidase deficiency. beta Glucuronidase activity was undetectable in affected cat fibroblasts and restored by retroviral gene transfer of rat beta-glucuronidase cDNA. beta-Glucuronidase mRNA was normal in affected cat testis by Northern blot analysis. Normal feline beta-glucuronidase cDNA was cloned and characterized, and amplified from affected cat fibroblasts by reverse transcription coupled polymerase chain reaction. There was a G-to-A transition in the affected cat cDNA that predicted an E351K substitution, destroyed a BssSI site, and eliminated GUSB enzymatic activity in expression studies. Multiple species comparison and the crystal structure of human beta-glucuronidase indicated that E351 is a highly conserved residue most likely essential in maintenance of the enzyme's conformation. BssSI digestion of polymerase chain reaction products amplified from genomic DNA indicated that affected cats were homozygous and cats with half-normal beta-glucuronidase activity were heterozygous for the missense mutation. Carriers identified in this manner produced affected kittens in prospective breedings, and a feline MPS VII breeding colony has been established. PMID- 10366444 TI - Genetic modifiers of polycystic kidney disease in intersubspecific KAT2J mutants. AB - Polycystic kidney disease (PKD) is a genetically heterogeneous disorder. In addition to the many PKD-causative loci mapped in mouse and human, a number of reports indicate that modifier loci greatly influence the course of disease progression. Recently we reported a new mouse mutation, kat2J, on chromosome (Chr) 8 that causes late-onset PKD and anemia. During the mapping studies it was noted that the severity of PKD in the mutant (C57BL/6J-kat2J/+ x CAST/Ei)F2 generation was more variable than that in the parental C57BL/6J strain. This suggested that genetic background or modifier genes alter the clinical manifestations and progression of PKD. Genome scans using molecular markers revealed three loci that affect the severity of PKD. The CAST-derived modifier on Chr 1 affects both kidney weight and hematocrit. The CAST-derived modifier on Chr 19 affects kidney weight, and the C57BL/6J-derived modifier on Chr 2 affects hematocrit. Additional modifier loci are noted that interact with and modulate the effects of these three loci. The mapping of these modifier genes and their eventual identification will help to uncover factors that can delay disease progression. These, in turn, could be used to design suitable modes of therapy for various forms of human PKD. PMID- 10366445 TI - A complete physical contig and partial transcript map of the Williams syndrome critical region. AB - Williams syndrome (WS) is a contiguous gene syndrome caused by hemizygosity for a chromosomal deletion at 7q11.23. The range of phenotypes includes mental retardation, dysmorphic facies, heart abnormalities, short stature, a specific cognitive profile, hyperacusis, and infantile hypercalcaemia. To identify all the deleted genes, we have constructed a detailed physical map and complete BAC/PAC contig of the critical region, extending a distance of approximately 2 Mb and delimited by the nondeleted markers D7S1816 and D7S489A. Somatic cell hybrids of WS patients were made and used to define the centromeric and telomeric deletion breakpoints, enabling the size of the WS deletion to be defined as approximately 1.4 Mb. Genes previously mapped to the region have been located on the contig, and we have isolated eight transcripts, two of which have been characterized as the genes CPETR1 and CPETR2. This contig and expressed sequence map will form the basis for the construction of a complete transcription map of the deleted region and will enable genotype-phenotype correlations to be attempted to identify the individual components of WS. PMID- 10366446 TI - Novel proteins interacting with the leucine-rich repeat domain of human flightless-I identified by the yeast two-hybrid system. AB - The flightless-I gene encodes a member of the gelsolin-like family of actin binding proteins linked to a leucine-rich repeat (LRR) domain. It is required for cellularization during early embryogenesis and normal development of the indirect flight muscles in Drosophila melanogaster. Although the association between actin and the gelsolin-like domain of the human Flightless-I homologue (FLI) has been established, its biological role is unknown. The human FLI gene is mapped within the Smith-Magenis microdeletion region of chromosome 17. We report the identification of two related genes, LRRFIP1 and LRRFIP2, encoding proteins that interact with the LRR domain of human FLI using the yeast two-hybrid system. LRRFIP1 exhibits sequence identity with the TRIP RNA-binding protein and GCF-2 transcriptional repressor, which are also related to the murine FLAP-1 gene. LRRFIP2 is a novel gene that shares sequence homology with LRRFIP1 and FLAP-1. LRRFIP1 and LRRFIP2 both express alternative splice variants in heart and skeletal muscle tissue. A coiled-coil domain, conserved within each encoded protein, serves as a potential interaction motif for FLI LRR. The occurrence of multiple proteins able to interact with FLI within the same tissue suggests that they may compete for the same binding site. Sequencing and PCR-directed genomic analysis indicate that LRRFIP1 and LRRFIP2 are related genes that arose from gene duplication. PMID- 10366447 TI - cDNA cloning, genomic structure, and chromosomal localization of a novel murine epidermis-type lipoxygenase. AB - Using a combination of degenerate PCR technique and conventional screening procedures, we isolated a cDNA encoding a novel lipoxygenase, termed epidermis type lipoxygenase-3 (e-LOX-3, gene symbol Aloxe3), from mouse skin. Aloxe3 mRNA is expressed in the stratified epithelia of skin, tongue, and forestomach. The cDNA encodes a protein of 711 amino acids with a calculated molecular mass of 80.6 kDa. The amino acid sequence shows approximately 54% identity to the recently identified 12(R)-lipoxygenase. Sequence comparison revealed a segment of 41 amino acid residues localized near the boundary between the N- and the C terminal domain sequences of the molecule, a structural feature that is also characteristic of 12(R)-lipoxygenase, suggesting that these two epidermis-derived lipoxygenases may be members of a novel structural class of mammalian lipoxygenases. The novel lipoxygenase gene is divided into 15 exons and 14 introns, spanning 22.3 kb of genomic DNA. By interspecific backcross analysis, the novel gene was localized to the central region of mouse chromosome 11. PMID- 10366448 TI - The third member of the transforming acidic coiled coil-containing gene family, TACC3, maps in 4p16, close to translocation breakpoints in multiple myeloma, and is upregulated in various cancer cell lines. AB - We have recently identified a novel gene, TACC1 (transforming acidic coiled coil containing gene 1), which is located close to FGFR1 within a region amplified in breast cancer on human chromosome 8p11. The coiled coil domain of this gene identified a series of cDNAs in the expressed sequence tag database, which suggested the existence of a family of TACC genes comprising at least three family members. We have now characterized the human and mouse TACC3 cDNAs, and demonstrate that this gene is upregulated in various cancer cell lines, and at Embryonic Day 15 in mice, suggesting that the TACC3 protein is involved in the control of cell growth and differentiation. The TACC3 gene maps telomeric to the FGFR3 gene in 4p16.3, close to a region disrupted by translocation breakpoints associated with multiple myeloma. Thus, TACC1, TACC2, and TACC3 map close to the corresponding FGFR1, FGFR2, and FGFR3 genes. The phylogenetic relationship among the three TACC genes is similar to that of the three FGFR family members. These relationships suggest that the FGFR and TACC genes arose from a physically linked ancestral gene pair. Subsequently, this gene pair has undergone two successive rounds of gene duplication to give rise to the three FGFR/TACC gene pairs on chromosomes 4, 8, and 10. PMID- 10366449 TI - Identification of a human homolog of the Drosophila rotated abdomen gene (POMT1) encoding a putative protein O-mannosyl-transferase, and assignment to human chromosome 9q34.1. AB - We have isolated a human gene homologous to Drosophila melanogaster rotated abdomen, rt, a poorly viable recessive mutation causing a clockwise twisted abdomen in affected flies due to defects in embryonic muscle development. The human gene, like rt, encodes a protein with high homology to the yeast mannosyl transferases (Pmts) and has been named POMT1. POMT1 is expressed as a 3.1-kb transcript in all tissues tested, with highest levels in testis and fetal brain. Alternative splicing of several exons in all tissues predicts the generation of several protein isoforms. The most common mRNA variant encodes a 725-aa protein with 40% identity and 62.5% similarity to rt, as well as 30.5% identity and 54% similarity to yeast Pmts. Computer prediction of protein sorting suggests that the POMT1 product could be an integral protein of the endoplasmic reticulum membrane. Given the strong conservation of protein motifs between POMT1 and the yeast Pmts, POMT1 may function as a mannosyl-transferase involved in O mannosylation of proteins, being the first of such a class found in mammals. The POMT1 locus has been assigned to human chromosome 9q34.1 by somatic cell hybrids, radiation hybrids, and linkage analysis. On the basis of the rt phenotype, POMT1 could be a candidate for uncharacterized genetic disorders of the muscular system, such as some forms of congenital muscular dystrophy or congenital myopathy. PMID- 10366450 TI - Characterization of MAD2B and other mitotic spindle checkpoint genes. AB - Aneuploidy is a characteristic of the majority of human cancers, and recent work has suggested that mitotic checkpoint defects play a role in its development. To further explore this issue, we isolated a novel human gene, MAD2B (MAD2L2), which is homologous to the spindle checkpoint gene MAD2 (MAD2L1). We determined the chromosomal localization of it and other spindle checkpoint genes, including MAD1L1, MAD2, BUB3, TTK (MPS1L1), and CDC20. In addition, we resolved the genomic intron-exon structure of the human BUB1 gene. We then searched for mutations in these genes in a panel of 19 aneuploid colorectal tumors. No new mutations were identified, suggesting that genes yet to be discovered are responsible for most of the checkpoint defects observed in aneuploid cancers. PMID- 10366451 TI - Integrated STS/YAC physical, genetic, and transcript map of human Xq21.3 to q23/q24 (DXS1203-DXS1059). AB - A map has been assembled that extends from the XY homology region in Xq21.3 to proximal Xq24, approximately 20 Mb, formatted with 200 STSs that include 25 dinucleotide repeat polymorphic markers and more than 80 expressed sequences including 30 genes. New genes HTRP5, CAPN6, STPK, 14-3-3PKR, and CALM1 and previously known genes including BTK, DDP, GLA, PLP, COL4A5, COL4A6, PAK3, and DCX are localized; candidate loci for other disorders for which genes have not yet been identified, including DFN-2, POF, megalocornea, and syndromic and nonsyndromic mental retardation, are also mapped in the region. The telomeric end of the contig overlaps a yeast artificial chromosome (YAC) contig from Xq24-q26 and with other previously published contigs provides complete sequence-tagged site (STS)/YAC-based coverage of the long arm of the X chromosome. The order of published landmark loci in genetic and radiation hybrid maps is in general agreement. Combined with high-density STS landmarks, the multiple YAC clone coverage and integrated genetic, radiation hybrid, and transcript map provide resources to further disease gene searches and sequencing. PMID- 10366452 TI - An in situ study of variant telomeric repeats in human chromosomes. AB - Variant telomeric repeats are selectively detected in human telomeres in situ by the novel approach of dideoxy-PRINS, displaying their organization in a format where all the individual chromosome ends can be viewed individually and simultaneously. All human chromosome ends are found to contain variant repeats, though not all types of repeats can be detected on all chromosome ends. Although the staining frequency at particular chromosome ends seems polymorphic among individuals, some chromosome ends are more commonly stained with a given probe than others. A few chromosome ends also appear with particularly strong signals. With a probe for one type of variant repeat ((AGGGTG)n), peculiar patterns with more than two signals per chromosome end are observed. PMID- 10366453 TI - A sequence-ready map of the human chromosome 17p telomere. AB - A half-YAC clone derived from human chromosome 17p was mapped at high resolution using cosmid subclone fingerprint analysis. Colinearity of the half-YAC with the telomeric human genomic DNA fragment was ascertained by RecA-assisted restriction endonuclease cleavage mapping. Previously isolated and radiation hybrid-mapped markers TEL17P37, TEL17P49, and TEL17P80 mapped 30-60 kb from the 17p terminus. This sequence-ready map permits high-resolution integration of genetic maps with the DNA sequences directly adjacent to the tip of human chromosome 17p, and will provide the cloned DNA required for ascertaining the nucleotide sequence of this subtelomeric region. PMID- 10366454 TI - Generation of a zebrafish P1 artificial chromosome library. AB - We have constructed a genomic P1 artificial chromosome library from the zebrafish. The library has been arrayed and archived in two hundred seventy-one 384-well microtiter dishes. It encompasses four to five genome equivalents with an average insert size of approximately 115 kb and is readily accessible to the scientific community. The library has been used by numerous investigators in the community and shown to be a useful reagent for chromosomal walking and positional cloning. PMID- 10366456 TI - How soon is it safe to undertake pregnancy after trophoblastic tumor? PMID- 10366455 TI - Mapping the mouse otoconin-90 (Oc90) gene to chromosome 15. PMID- 10366457 TI - Outcome of pregnancies occurring before completion of human chorionic gonadotropin follow-up in patients with persistent gestational trophoblastic tumor. AB - OBJECTIVE: To determine the outcome of pregnancies occurring before completion of human chorionic gonadotropin follow-up in patients treated with chemotherapy for gestational trophoblastic tumor. METHODS: Retrospective record review of patients with gestational trophoblastic tumor who conceived before standard hCG follow-up was completed during 1973-1998. RESULTS: Forty-three patients treated for gestational trophoblastic tumors conceived before human chorionic gonadotropin follow-up was completed. The antecedent pregnancy was complete mole in 31 (72.1%) and partial mole in 12 (27. 9%) patients. Of the 43 patients, 39 (90.7%) had stage I, 1 had stage II, and 3 had stage III disease. The mean interval from human chorionic gonadotropin remission to new pregnancy was 6.3 months (range 1 11 months). Ten patients underwent elective termination and four patients were lost to follow-up. Of the remaining 29 patients, 22 (75.9%) had term live births, 3 (10.3%) had preterm delivery, 3 had spontaneous abortion, and 1 (3.5%) had a repeat mole. Two cases of fetal anomalies were detected; one was inherited polydactyly and the other was hydronephrosis. One patient developed choriocarcinoma with lung involvement and underwent cesarean section at 28 weeks; a normal fetus was delivered and no choriocarcinoma was detected in the placenta. CONCLUSION: Pregnancies occurring in patients treated for gestational trophoblastic tumor before standard human chorionic gonadotropin follow-up is completed may continue under close clinical surveillance since the majority have a favorable outcome. However, patients should also be advised of the low but important risk of delayed diagnosis in case tumor relapse develops during early subsequent pregnancy. PMID- 10366458 TI - Adenocarcinoma in situ of the cervix: management and outcome. AB - OBJECTIVE: The aim of this study was to review the management and outcome of patients with adenocarcinoma in situ of the cervix and to evaluate the significance of endocervical cone margin status in these patients. METHODS: A retrospective review of records between January 1988 and December 1996 identified 40 patients with adenocarcinoma in situ on cone biopsy for whom complete information was available. The median follow-up was 38 months. RESULTS: The mean age was 37 years, and the mean parity was 1.3. Fifty-three percent of the patients had prior abnormal cervical cytology. The initial Pap smear that led to the patient's referral was abnormal in 39 (98%). Initial cervical biopsies showed adenocarcinoma in situ and/or glandular dysplasia in 28 (70%), squamous dysplasia in 2 (5%), chronic inflammation in 2 (5%), and no pathologic changes in 2 (5%) patients. Initially no biopsies were performed in 3 (7.5%) patients and the results of 3 (7.5%) biopsies were unknown. Subsequently, all patients had cone biopsies. The endocervical margins were positive for glandular abnormalities in 24% of cold knife cones (CKC), 75% of LEEPs, and 57% of laser cones. The ectocervical margins were positive for squamous and/or glandular abnormalities in 8% of CKCs, 13% of LEEPs, and 57% of laser cones. ECCs above the cone were obtained in 28 patients, and only 1 (3%) was positive. The definitive treatment was hysterectomy in 27, repeat cone in 5, and no additional therapy in 8 patients. The pathology showed residual disease in 44% of treated patients. From 16 cone biopsies with negative margins who had subsequent treatment, there was residual disease in 5 (31%) specimens (1 adenocarcinoma in situ, 1 mild glandular dysplasia, 3 glandular atypia). From 16 cones with positive margins who had subsequent treatment, there was residual disease in 9 (56%) specimens. The patients with negative ECCs above the cone regardless of margin status had residual disease in 58% of treated specimens. CONCLUSION: Women with adenocarcinoma in situ of the uterine cervix had residual disease in 31% of cases with negative margins in cone biopsies and/or with negative ECCs and in 56% of cases with positive endocervical margins. LEEP cones had higher rate of positive endocervical margins (75%) compared to CKC (24%) and laser cone (57%). If maintaining reproductive capacity is desired, we would recommend CKC; however, this does not guarantee absence of the disease. PMID- 10366459 TI - The significance of Erb-b2 immunostaining in cervical cancer. AB - OBJECTIVE: The aim of this study was to assess the relationship between survival and erb-b2 immunohistochemical staining in patients with early stage cervical carcinoma. METHODS: Archival specimens for 126 patients with stage IB/IIA cervical carcinoma treated with radical hysterectomy and bilateral pelvic node dissection (RH-BPND) were retrieved and submitted to immunohistochemistry for ERBB2 expression. The association between positive results and poor survival was assessed in a multivariate analysis. RESULTS: Erb-b2 immunostaining was significantly associated with poor survival (P = 0.0284) but less so than parametrial extension (P = 0.0014) and nodal disease (P = 0.0106). Tumor type (squamous/adenosquamous/adenocarcinoma) and the status of surgical margins were not significantly associated with survival. CONCLUSIONS: These results supported further investigations of ERBB2 expression as a marker of high-risk disease in patients treated with RH-BPND. PMID- 10366460 TI - An association between LSIL and the high secretor phenotype of IL-1beta. AB - OBJECTIVE: The aims of this study were to determine the frequency of a nucleotide transition from C to T, which leads to increased transcription of interleukin 1beta (IL-1beta) in patients with different grades of cervical lesions, and to determine whether a correlation exists between the genotypes and cervical lesions. METHODS: One hundred forty-seven DNA samples from patients with different grades of cervical lesions were compared with 100 healthy, age and sex matched bone marrow donors. TaqI restriction digest of PCR products was used to analyze the IL-1beta +5887 C --> T mutation and the results were confirmed using induced heteroduplex analysis with an induced heteroduplex generator. RESULTS: The Pearson chi2 test (Yate's correction) was used for statistical analysis. Patients with LSIL (n = 80) demonstrated a high frequency of allele T, previously associated with high IL-1beta secretor phenotype, compared to controls (P = 0.000012). A trend was also observed in patients with HSIL (n = 28, P = 0.039). CONCLUSIONS: We report for the first time a highly significant association between high secretor IL-1beta phenotypes (i.e., IL-1beta +5887 CT or TT genotypes) and LSIL. A less significant association exists with HSIL. The intrinsic ability to produce variable amounts of IL-1beta during different clinical stages involving cervical lesions may be of immunological importance in their pathogenesis. PMID- 10366461 TI - Beta1-integrins partly mediate binding of ovarian cancer cells to peritoneal mesothelium in vitro. AB - Ovarian cancer cells frequently metastasize by implanting onto the peritoneal mesothelial surface of the abdominal cavity. Although the CD44 molecule expressed by ovarian cancer cells is known to partly mediate this process, the role of other adhesion proteins such as beta1-integrins has been previously difficult to demonstrate using the 4B4 anti-beta1 neutralizing antibody. In view of the widespread expression of beta1-integrins in ovarian cancer, however, we have further examined the potential role of this class of molecules in ovarian cancer cell implantation through the use of an alternative anti-beta1 neutralizing antibody, MAB13. We now report that MAB13 is capable of inhibiting the binding of three separate human ovarian cancer cell lines (36M2, CAOV-3, SKOV-3) to mesothelium (mean 37 +/- 4% inhibition, n = 21, P < 0.001). An additive inhibitory effect was observed when MAB13 was combined with anti-CD44 antibody (clone 515) (mean 63 +/- 3% inhibition, n = 19, P < 0.001), suggesting that binding occurs through two independent pathways involving both beta1-integrins and CD44. Because fibronectin is an extracellular matrix ligand recognized by many types of integrins and is abundantly expressed on mesothelial cells, the inhibitory effects of MAB13 and 4B4 on ovarian cancer cell binding to fibronectin were directly compared. MAB13 inhibited ovarian cancer cell binding to fibronectin to a significantly greater degree than did 4B4, suggesting that the discordant effects of these two antibodies on mesothelial adhesion may be partly related to their differential ability to neutralizing fibronectin-mediated binding. Studies using anti-alpha5 neutralizing antibody demonstrated that the alpha5beta1 fibronectin receptor contributes to approximately 50% of integrin mediated binding of 36M2 and CAOV-3 cells (which express the alpha5 chain in 54 and 58% of cells, respectively). Since the RGD sequence of fibronectin is a known recognition site for many types of integrins, including alpha5beta1 and alphanubeta3, we performed binding in the continued presence of both anti-alpha5 and RGD peptide in order to exclude other types of fibronectin-integrin interactions. The addition of RGD peptides at concentrations known to be capable of blocking fibronectin binding resulted in no additional inhibitory effect over that observed with anti-alpha5 antibody alone, suggesting that alpha5beta1 was the major receptor responsible for fibronectin-mediated ovarian cancer binding to mesothelium. These data demonstrate that ovarian cancer cell binding to peritoneal mesothelium is mediated by several adhesion pathways and that simultaneous inhibition of both beta1-integrin and CD44 function may be necessary in order to significantly limit the intraabdominal spread of this tumor in vivo. PMID- 10366462 TI - Cell cycle arrest in endometrial carcinoma cells exposed to gonadotropin releasing hormone analog. AB - Gonadotropin-releasing hormone (GnRH) has been shown to have an inhibitory effect on the growth of several hormone-dependent human tumors. We have treated a human endometrial cancer cell line which expresses GnRH receptor with GnRH analog, D Trp6-LHRH, in order to study whether there are differences in cell cycle kinetic response. Flow cytometric analysis revealed that cultured carcinoma cells showed a cell cycle arrest at the G1-S transition after treatment with 10 microM D-Trp6 LHRH for 36 h. Western blot analysis showed that the level of p16 protein was obvious following 24 h of D-Trp6-LHRH treatment. These results suggest that the mechanism by which GnRH inhibits the growth of endometrial carcinoma cells may include effects on cell cycle arrest. PMID- 10366463 TI - MMP-2 and TIMP-2 expression correlates with poor prognosis in cervical carcinoma- a clinicopathologic study using immunohistochemistry and mRNA in situ hybridization. AB - OBJECTIVE: The spread of malignant neoplasms is closely associated with matrix and basement membrane degradation, mediated by various classes of proteolytic enzymes. Matrix metalloproteinases (MMP) appear to have a key role in the sequence of events that lead to local invasion and metastasis. The present study evaluated the role of matrix metalloproteinase-2 (MMP-2), tissue inhibitor of metalloproteinases-2 (TIMP-2), and membrane-type metalloproteinase (MT1-MMP) in cervical neoplasia. METHODS: We have analyzed 49 uterine cervical squamous cell carcinomas, 10 cases of high-grade cervical intraepithelial neoplasia (CIN II III), and 10 control cervices for the presence of MMP-2, TIMP-2, and MT1-MMP using in situ hybridization. MMP-2 protein expression was evaluated using immunohistochemistry. Results were analyzed for possible correlation with disease outcome. RESULTS: MMP-2, TIMP-2, and MT1-MMP mRNA were localized to both stromal and tumor cells. However, an intense signal for MMP-2 was detected almost exclusively in tumor cells and was uniformly absent from CIN lesions and control cervices. Conversely, intense signals for TIMP-2 and MT1-MMP were detected in both stromal and tumor cells of invasive carcinomas, more often for the former. As with MMP-2, they were absent from CIN lesions. MMP-2 protein expression was enhanced in tumor cells compared to CIN cases and controls, significantly compared to the latter (P = 0.01). The presence of both MMP-2 and TIMP-2 mRNA in tumor cells correlated with advanced stage (P = 0.003 for MMP-2, P = 0.002 for TIMP-2) and with poor survival (P = 0.003 for MMP-2, P = 0.002 for TIMP-2) in univariate analysis. In addition, their presence in tumor cells intercorrelated (P = 0.002). In multivariate survival analysis, MMP-2 presence retained its association with survival (P = 0.004), in addition to patient age (P = 0.027) and advanced stage (P = 0. 0002). CONCLUSIONS: Both MMP-2 and TIMP-2 have a key role in extracellular matrix invasion in cervical carcinoma, largely through their elaboration by tumor cells. The presence of mRNA for both proteins is interrelated and is associated with poor survival. PMID- 10366464 TI - Laparoscopic lymphadenectomy for gynecologic malignancies. AB - OBJECTIVE: The purpose of our study was to detail our 5-year experience with laparoscopic lymphadenectomy for gynecologic malignancies. METHODS: From 11/5/92 to 3/9/98, we performed laparoscopic lymphadenectomies on 94 patients with various gynecologic malignancies. Pelvic, paraaortic, and combinations of both pelvic and paraaortic lymphadenectomies were performed depending on the primary site of disease and indication for lymph node dissection. Data were prospectively collected on all patients. RESULTS: From 11/5/92 to 3/9/98 we performed 94 laparoscopic lymphadenectomies for gynecologic malignancies. The distribution included 64 patients with cervical cancer, 14 with ovarian cancer, 12 with endometrial cancer, 2 with fallopian tube cancer, 1 with a uterine malignant mixed mesodermal tumor, and 1 with a metastatic neuroendocrine tumor. Fifty-five patients had only pelvic lymph node dissections, 9 patients had paraaortic dissections only, and 30 had both pelvic and paraaortic dissections performed. Among 30 patients having laparoscopic lymphadenectomy only, the mean hospital stay was 3.6 days. Included in this group were 19 patients who received postoperative neoadjuvant chemotherapy for cervical cancer as inpatients prior to ambulatory radiation therapy. The mean length of stay for this group was 4.6 days versus 1.7 days for the 11 patients who did not receive postoperative chemotherapy (P = 0.0025). The mean number of pelvic nodes was 11.9 (range 0-57), with a mean of 4. 5 between 11/5/92 and 12/31/95 and a mean of 19.1 from 1/1/96 to 3/9/98. The mean number of paraaortic nodes obtained was 3.7 (range 0-14), with a mean of 3.4 from 11/5/92 to 12/31/95 and a mean of 4.1 from 1/1/96 to 3/9/98. A total of 3 patients required conversions to laparotomy. One was for a vascular injury to the vena cava, 1 for a large tumor extending to both sidewalls, and the third for removal of densely matted lymph nodes. CONCLUSIONS: Laparoscopic lymphadenectomy is a technically feasible procedure for patients with gynecologic malignancies requiring lymph node dissections, with an acceptable safety profile and nodal yield. The number of nodes obtained increased in direct proportion to operator experience. In addition, patients may benefit from a decrease in hospital stay compared to conventional lymphadenectomy via laparotomy. PMID- 10366465 TI - Results of primary and adjuvant radiotherapy in the treatment of mixed Mullerian tumors of the corpus uteri. AB - OBJECTIVE: The benefit of primary or adjuvant irradiation in the treatment of mixed Mullerian tumors is still not clear. METHODS: During 1981-1997 63 patients were referred for primary (n = 13) or postoperative (n = 50) radiotherapy. Analysis of outcome of primarily and postoperatively irradiated patients was performed separately because of different staging systems. Of 50 patients treated after surgery 29 presented in histopathologic stage I, 4 in stage II, 14 in stage III, and 3 in stage IV. Clinical stage distribution for primary treatment was stage I: n = 9, stage II: n = 1, stage III: n = 3. Forty-four patients in the postoperatively treated group and 6 in the primarily treated group received radiotherapy with a curative intent; external beam therapy was given up to 56 Gy to the pelvis combined with intravaginal or intracavitary brachytherapy. RESULTS: Five-year actuarial overall survival, disease-specific survival, local control, and distant control for 50 patients receiving adjuvant irradiation was 52.9, 57. 5, 83.4, and 70.8%, in stage I: 68.4, 76.1, 95.2, and 81.7%, in stage II: 50.0, 50.0, 75.0, and 66.7%, and in stage III: 31.3, 34.1%, 70.4, and 47.6%, respectively. Four of 13 patients treated with primary irradiation achieved long term local control. CONCLUSION: These data suggest that adjuvant radiotherapy improves local control and disease specific survival in the treatment of mixed Mullerian tumors compared to data in the literature concerning treatment by surgery alone. PMID- 10366466 TI - Vascular endothelial growth factor expression as a prognostic index in serous ovarian cystoadenocarcinomas: relationship with MIB1 immunostaining. AB - OBJECTIVE: The aim of our study was to investigate the expression of vascular endothelial growth factor (VEGF) by neoplastic cells in serous ovarian cystoadenocarcinomas; the correlation between this marker of angiogenesis, histopathologic parameters, disease-free survival, and MIB1 immunostaining was also evaluated. MATERIALS AND METHODS: Thirty-two patients with serous ovarian cystoadenocarcinoma, treated at the Institute of Gynecology and Obstetrics, Ancona University (Italy), were used as study population; 10 women with serous cystoadenoma were also analyzed. The expression of VEGF was immunohistochemically evaluated by polyclonal antibody anti-VEGF (Santa Cruz, CA, dilution 1:100) on formalin-fixed paraffin-embedded tissue. RESULTS: Compared to cystoadenomas, the tissutal VEGF immunostaining was significantly higher in cystoadenocarcinomas, with the highest values in architectural grade 3 neoplasms (P < 0. 001). A direct relationship was observed between VEGF immunostaining and MIB1 index (r = 0.44, P = 0.013). A relationship was defined between VEGF expression and disease-free survival, evaluated by Cox hazards analysis (P < 0.001). CONCLUSIONS: Angiogenesis, evaluated by VEGF immunostaining, seems to be an interesting prognostic indicator in serous ovarian cystoadenocarcinoma, involved in neoplastic proliferation. PMID- 10366467 TI - Ovarian histopathology in breast cancer patients receiving tamoxifen. AB - OBJECTIVE: The purpose of this study was to examine ovarian histopathology in tamoxifen-treated breast cancer patients undergoing oophorectomy. METHODS: We reviewed the records and ovarian histopathology of 152 breast cancer patients who underwent oophorectomy at a single institution between January 1980 and October 1996. At the time of oophorectomy, 99 patients had never received tamoxifen, 44 patients were currently receiving tamoxifen, and 9 patients had previously received tamoxifen. Patient demographic and medical data and indication for oophorectomy were examined. Ovarian histopathology was classified as normal, functional ovarian cyst, benign ovarian tumor, endometriosis, ovarian cancer, and metastatic cancer. RESULTS: Patient characteristics and indication for oophorectomy did not differ significantly based on tamoxifen exposure. There was no difference in the occurrence of benign ovarian tumors, functional ovarian cysts, or metastatic breast cancer based on tamoxifen exposure. Tamoxifen-treated patients were less likely to have ovarian cancer, 0 of 53 patients (95% confidence interval (CI): 0.0%, 6.7%) compared with 10 of 99 patients (95% CI: 5.0%, 17.8%) patients not receiving tamoxifen (P = 0.015). Endometriosis was slightly more common in patients currently receiving tamoxifen, but the difference was not statistically significant. CONCLUSIONS: In women undergoing oophorectomy, there was no evidence that tamoxifen exposure was associated with an increase in benign or malignant primary or metastatic ovarian neoplasm or in functional ovarian cysts. Further study is necessary to better define any association between tamoxifen and endometriosis and the effect of tamoxifen on ovarian cancer risk. PMID- 10366468 TI - Predictors of recurrence in surgical stage II endometrial adenocarcinoma. AB - OBJECTIVE: A retrospective review of surgical stage II endometrial carcinoma was performed to evaluate clinical course, treatment, recurrence rate, and survival. METHODS: A list of patients with clinical and surgical stage II endometrial carcinoma was obtained through the tumor registry and from the pathology department from 1988 to 1996. Data were collected on all cases of patients with endometrial carcinoma meeting stage II criteria by FIGO surgical staging. Variables including stage, histology, grade, lymph vascular space invasion (LVI), type and extent of surgery, radiation type and amount, smoking, menstrual status, parity, and age were evaluated for their predictive ability of disease recurrence. Cox proportional hazard regression models were used to examine the potential predictors of time to relapse univariately and multivariately. RESULTS: Of patients identified, 65 underwent primary surgical staging. Only adenocarcinomas were included. Mean follow-up time was 4.7 years (range 0.2-9.6 years). Postoperative radiation was given to 85.7% of patients. There were 10 patients (15.4%) with recurrence of disease with a mean time to recurrence of 25 months. Five-year disease-specific survival was 93%. The only significant predictor of time to relapse was LVI (P = 0.002) in the multivariate analysis. CONCLUSION: This retrospective review suggests that primary surgery followed by postoperative radiation therapy gives excellent results in surgical stage II disease. LVI appears to be a strong predictor of disease recurrence regardless of postoperative radiation therapy. It is difficult to draw conclusions about the type and amount of radiation given because recurrence rate is so low; however, it is reasonable to continue adjuvant radiation especially in cases where LVI is identified. PMID- 10366469 TI - Prospective trial of aggressive postoperative bowel stimulation following radical hysterectomy. AB - INTRODUCTION: Postoperative traditional feeding protocols are not based on scientific studies, but rather on anecdotal evidence. We present the first prospective trial of aggressive postoperative bowel stimulation following radical hysterectomy in an attempt to determine its effect on the length of hospital stay. METHODS: Twenty consecutive patients undergoing radical hysterectomy were entered onto a prospective trial of aggressive postoperative bowel stimulation, which consisted of 30 cc milk of magnesia p.o. b.i.d. starting on postoperative day 1 and biscolic suppositories q.d. starting on day 2. A clear liquid diet was begun following flatus or bowel movement and patients were discharged 12 h after tolerating a clear liquid diet. Diet was slowly advanced at home. RESULTS: Median time to flatus was 3 days, bowel movement 3 days, and clear liquid diet 3 days. Median time to discharge was 4 days. No patients developed ileus or bowel obstructions and there were no readmissions for bowel complications. Our median time to discharge of 4 days represents a 50% reduction in hospital stay compared to our previous prospective study using traditional postoperative bowel management (8 days), which was statistically significant at P = 0.001. CONCLUSION: Aggressive bowel stimulation with milk of magnesia and biscolic suppositories resulted in early return of bowel function and early discharge with no noticeable complications. PMID- 10366470 TI - Expression of tenascin in human cervical cancer--association of tenascin expression with clinicopathological parameters. AB - OBJECTIVE: Tenascin is an extracellular matrix glycoprotein, relevant for embryonal and fetal development, which is reexpressed in the stroma of benign and malignant tumors. Little is known about the molecular interaction of tenascin during neoplastic transformation and tumor progression in cervical cancer. METHOD: We studied the expression of tenascin in normal tissue of the cervix uteri, cervical carcinoma in situ, and invasive cervical carcinoma in paraffin sections by immunohistochemistry using a monoclonal antibody. Tenascin immunoreactivity was compared with various prognostic parameters. RESULTS: In normal cervical tissue (n = 5) and in cervical carcinoma in situ (n = 10) only vessel walls showed a weak tenascin cross-reactivity, whereas tenascin was not expressed in the epithelial layer or the underlying connective tissue. In invasive cervical carcinoma (n = 89) tenascin expression was markedly increased. In 84% (n = 75) of the cases examined a strong tenascin immunoreactivity was noted around and within the tumor cell nests. Sixteen percent (n = 14) of infiltrating cervical carcinomas showed no tenascin immunoreactivity. A definite correlation was found between weak or no tenascin expression and slight desmoplastic mesenchymal reactivity (n = 42/91%, P < 0.001), lymphatic space invasion (n = 54/81%, P < 0.001), and lymph node metastases (n = 30/77%, P < 0.05). Tenascin-positive patients had a significantly better prognosis than tenascin-negative patients (mean survival time of 56.5 +/- 4.1 months versus 31.9 +/- 5.6 months, P < 0.05). CONCLUSION: Based on these findings we discuss that the appearance of tenascin is an indicator of an adequate biological defense in cervical cancer patients. The tenascin staining may therefore be useful for detecting a subgroup of invasive cancer patients missing tenascin reactivity with alterations of stromal defense and a poorer prognosis. PMID- 10366471 TI - Persistent chemosensitivity to platinum and/or paclitaxel in metastatic endometrial cancer. AB - It has long been recognized that individuals with ovarian cancer who initially respond to a platinum-containing chemotherapeutic regimen may exhibt a second response to platinum (cisplatin or carboplatin) at the time of recurrence. In this report, we describe three individuals with metastatic endometrial cancer who demonstrated secondary responses to platinum/paclitaxel-based regimens. Endometrial cancer should be added to the list of malignancies for which platinum and/or paclitaxel are considered as second-line treatment options in patients previously responding to the agents. PMID- 10366472 TI - Blindness as a consequence of a paraneoplastic syndrome in a woman with clear cell carcinoma of the ovary. AB - BACKGROUND: Paraneoplastic syndromes are rare conditions associated with cancer that result in serious disease states at unique sites. In 1982, a report of bilateral diffuse uveal melanocytic proliferation associated with nonocular cancers which resulted in blindness was reported. We present a case of a woman with recurrent ovarian cancer who developed this paraneoplastic syndrome. CASE: A 55-year-old woman had been diagnosed in 1990 with an ocular melanoma of her right eye and in 1994 with clear cell carcinoma of the ovary. With recurrence of ovarian cancer, new eye lesions were identified in both eyes. After enulcleation of her right eye, an ocular melanoma and diffuse bilateral melanocytic proliferation (BDUMP) were found. The sight in her left eye continued to deteriorate as other signs of BDUMP occurred in the eye. Within 1 month of diagnosis, the patient was blind. She subsequently succumbed to progression of ovarian cancer. CONCLUSION: Recurrent ovarian cancer is usually an intraabdominal disease that results in gastrointestinal dysfunction. This case illustrates a rare paraneoplastic syndrome associated with ovarian cancer that mimics metastatic disease to the eye, but has a different pathophysiology. PMID- 10366473 TI - Cancer-associated retinopathy in a patient with advanced epithelial ovarian carcinoma. AB - BACKGROUND: Paraneoplastic phenomena, such as retinopathy, may herald an unsuspected gynecologic malignancy. CASE: A 75-year-old woman presented to a neuro-ophthalmologist with abrupt onset of unilateral visual loss. A diagnosis of branch retinal artery occlusion was made and she was treated with aspirin. An echocardiogram subsequently revealed atrial dilation and she was placed on coumadin therapy. Her vision worsened and a cancer-associated retinopathy was entertained. A serum cancer-associated retinopathy antibody was detected; subsequent computed tomographies of the abdomen and pelvis revealed findings consistent with a primary ovarian carcinoma. CONCLUSION: Patients with unexplained ophthalmologic symptoms may harbor an underlying gynecologic cancer. PMID- 10366474 TI - Immature teratomas of the genital tract in older women. AB - We report the clinicopathologic features of three women, 40 years of age or older, with malignant genital tract immature teratomas. All had FIGO stage III, grade II or grade III tumors. One tumor arose from the fallopian tube, the second from the ovary, and the third involved the cortical surfaces of both ovaries with minimal parenchymal involvement. The tumors weighed 1700, 5660, and 330 g and had histologic features similar to those generally seen in younger women. Two of the women died within 1 year of diagnosis. Interval growth of tumor after treatment with chemotherapy was documented in the third patient; she was reexplored and all of the excised tumor was composed of mature tissues. These cases affirm that, although rare, malignant germ cell tumors can occur in older peri- or postmenopausal women. PMID- 10366476 TI - Primary angiosarcoma of the ovary: a case report and review of the literature. AB - INTRODUCTION: Gynecological sarcomas are rare and have a poor prognosis. Uterine sarcomas are most common accounting for 4% of all uterine tumors. Ovarian sarcomas are less frequent and are usually carcinosarcomas. CASE REPORT: A previously healthy 40-year-old G2P2 presented for evaluation of 72 h of right upper quadrant pain and shortness of breath. A malignant right pleural effusion, ascites, and adnexal mass were found. Surgical staging and suboptimal debulking revealed pure angiosarcoma of the ovary Stage IV. DISCUSSION: There are 12 cases of ovarian angiosarcoma reported in the literature. Ten of these cases presented in advanced stages with survivals of 2-30 months. Various chemotherapy regimens have been tried on these tumors including the most recent recommendation of MAID (mesna, doxorubicin, ifosfamide, and dacarbazine) and prognosis remains poor. Our patient underwent elective right pleurodesis via video-assisted thorascopic surgery under local anesthesia for an early recurrent right pleural effusion and subsequently began MAID chemotherapy. PMID- 10366475 TI - Spontaneous rupture of the urinary bladder is not a rare complication of radiotherapy for cervical cancer: report of six cases. AB - BACKGROUND: Spontaneous intraperitoneal rupture of the urinary bladder is an extremely rare event. It has been reported to be a rare complication of radiation therapy for cervical cancer, but no studies have ever reported the incidence of this life-threatening complication. MATERIALS AND METHODS: From August 1981 through December 1988, 143 patients with carcinoma of the uterine cervix were treated with high-dose-rate intracavitary brachytherapy combined with external beam therapy at Kobe City General Hospital. RESULTS: Of these patients, three (2.1%) suffered spontaneous intraperitoneal rupture of the urinary bladder as a late complication of radiation therapy between August 1995 and February 1998. Three other patients, treated with radiation therapy for cervical cancer at other hospitals, were also admitted to our hospital with this complication between August 1995 and February 1998. All six patients underwent laparotomy and repair of the perforation. However, rerupture of the bladder occurred in three of these patients. CONCLUSION: Spontaneous intraperitoneal rupture of the urinary bladder after radiation therapy for cervical cancer is less rare than previously expected, and urologists must consider the possibility of occurrence of this life threatening event following radiation therapy. PMID- 10366477 TI - Malignant struma ovarii: two case reports and a review of the literature. AB - Struma ovarii consists of thyroid tissue derived from germ cells in a mature teratoma. Malignant transformation is very rare, with clinically evident metastatic disease reported in approximately 20 cases. The rarity of this disease renders evaluation of treatment modalities difficult. There is evidence that these tumors behave like their thyroid counterparts, and cytoreductive surgery followed by ablation with radioactive iodine has been advocated. We report the diagnosis and treatment of 2 patients with metastatic malignant struma ovarii treated with a combination of surgery and radiation therapy. PMID- 10366478 TI - Long-term consequences following conservative management of epithelial ovarian cancer in an infertile patient. AB - A 35-year-old woman with primary infertility underwent an ovarian cystectomy for a 5 x 4 cm left adnexal mass. There was no macroscopic evidence of metastatic disease. The final pathology report revealed a poorly differentiated serous cystadenocarcinoma. Because the patient desired to retain child-bearing capacity, she refused a surgical staging of her ovarian cancer. She elected to receive combination chemotherapy. This was then followed by a negative reassessment laparotomy. The patient was diagnosed with recurrent, metastatic ovarian carcinoma 10 years later. PMID- 10366479 TI - Radiotherapy for the treatment of metastatic granulosa cell tumor in the mediastinum: a case report. AB - OBJECTIVE: We report a case of metastatic ovarian granulosa cell tumor in the mediastinum with a 2-year disease-free interval after treatment with radiotherapy and review the literature regarding the use of radiotherapy in recurrent and metastatic granulosa cell tumor. METHODS: The patient's medical records, histological slides, and radiological films were reviewed. The pertinent references were obtained using a Medline search and cross-references. RESULTS: A patient with Stage 1A granulosa cell tumor developed a recurrence in the retroperitoneum 10 years after initial surgery. She was treated with chemotherapy followed by surgical resection. She subsequently developed metastatic tumor in the mediastinum which responded completely to radiotherapy. She has remained disease free for 2 years since the completion of radiotherapy. CONCLUSION: Radiotherapy is a treatment option that should be considered in localized recurrent or metastatic granulosa cell tumor that is not amenable to surgery as it can potentially control the disease for several years. PMID- 10366480 TI - Complete response of a stage IV uterine papillary serous carcinoma to neoadjuvant chemotherapy with Taxol and carboplatin. AB - Uterine papillary serous carcinoma (UPSC) is an aggressive histologic subtype of endometrial cancer. Currently, no effective chemotherapy regimens exist. We report a case of complete response of a stage IV UPSC to neoadjuvant chemotherapy with Taxol and carboplatin. PMID- 10366481 TI - A primary ovarian leiomyosarcoma with micro-invasive features (stage I): is surgical excision enough? PMID- 10366482 TI - Reply PMID- 10366483 TI - Surgical staging in endometrial cancer: is excellent survival due to thorough staging or patient selection? PMID- 10366484 TI - Reply PMID- 10366485 TI - Concerns with "maintenance chemotherapy" for patients with recurrent platinum sensitive ovarian cancer. PMID- 10366486 TI - Reply PMID- 10366487 TI - Coherent nature of the radiation emitted in delayed luminescence of leaves AB - After exposure to light, a living system emits a photon signal of characteristic shape. The signal has a small decay region and a long tail region. The flux of photons in the decay region changes by 2 to 3 orders of magnitude, but remains almost constant in the tail region. The decaying part is attributed to delayed luminescence and the constant part to ultra-weak luminescence. Biophoton emission is the common name given to both kinds of luminescence, and photons emitted are called biophotons. The decay character of the biophoton signal is not exponential, which is suggestive of a coherent signal. We sought to establish the coherent nature by measuring the conditional probability of zero photon detection in a small interval Delta. Our measurements establish the coherent nature of biophotons emitted by different leaves at various temperatures in the range 15-50 degrees C. Our set up could measure the conditional probability for Delta10-fold increase in the maximum rate of DNP-SG efflux. DNP-SG efflux in MRP1-expressing MCF7 cells was ATP-dependent and exhibited an apparent Km for DNP-SG of 95 microM. MRP1 expression alone, however, had no effect on DNP-SG formation. Combined expression of GST P1-1 and MRP1 increased the rates of DNP-SG formation when cells were exposed to 10 microM CDNB. Moreover, combined expression of GSTP1-1 with MRP1 moderately augmented MRP1-mediated resistance to CDNB but only during short term (10 min) exposures to CDNB where IC50 values were in the 8-10 microM range. In contrast, expression of GST P1-1 in the absence of MRP1 slightly sensitized cells to the toxicity of CDNB (10 min exposures), despite increasing rates of DNP-SG formation. The sensitization to CDNB in cells expressing GST P1-1 alone was associated with increased intracellular accumulation of DNP-SG, indicating that DNP-SG may contribute to CDNB toxicity. The potential toxicity of DNP-SG is also suggested by the finding that inhibition of DNP-SG formation by prior glutathione depletion confers resistance to CDNB cytotoxicity in MRP1-poor MCF7 cells. Altogether, our results demonstrate that glutathione conjugation and MRP1-mediated conjugate efflux can operate together to confer resistance to CDNB. The data indicate that MRP1-mediated conjugate efflux is required for cytoprotection from CDNB because its conjugate (DNP-SG), when present at high intracellular levels, may also be toxic to cells. PMID- 10366542 TI - Comparison of aldicarb and methamidophos neurotoxicity at different ages in the rat: behavioral and biochemical parameters. AB - Young organisms are often more sensitive to the toxic effects of pesticides, and this finding has spurred research on further characterization of this susceptibility. The neurotoxic effects of cholinesterase (ChE)-inhibiting pesticides are of particular concern for human health risk assessment due to the widespread exposure potential in children. This study evaluated age-related differences in susceptibility for a carbamate (aldicarb) and an organophosphorus pesticide (methamidophos). Comparisons were made between preweanling (Postnatal Day 17, PND17), postweanling (PND27), and adult (approximately PND70) male and female rats. All were acute studies using oral administration. Sensitivity was quantified by (1) determination of maximally-tolerated doses (MTDs); (2) measurement of brain and blood ChE inhibition; and (3) neurobehavioral evaluation using end points known to be sensitive indicators of exposure to anticholinesterases. MTD data showed that preweanling rats were twice as sensitive as adults to aldicarb, but there was no differential sensitivity to methamidophos. The dose-response data for brain ChE inhibition followed a similar pattern of age-related differences, and similar levels of inhibition were measured at the MTD regardless of age. Dose-response and time course studies of neurobehavioral end points indicated that differential effects due to age depend on the behavioral end point examined. Following aldicarb administration, the dose response curves for a few end points overlapped; however, the young rats otherwise showed fewer signs of toxicity than did the adults despite similar levels of brain ChE inhibition. Motor activity assessment showed that aldicarb did not produce any activity depression in PND17 rats, whereas the data for the PND27 and adult rats overlapped. With methamidophos, the dose-response curves for most end points for preweanling and adult rats were quite similar. Aldicarb induced ChE inhibition was readily reversible in all age groups, whereas with methamidophos, enzyme activity recovered more rapidly in the young. Most behavioral alterations had recovered by 24 h with either pesticide. The results of these studies indicate that (1) ChE-inhibiting pesticides are not all the same regarding relative sensitivity of the young; (2) age-related differences were reflected in both the MTDs and degree of ChE inhibition; and (3) age-related differences in neurobehavioral measures depended on the pesticide and on the end points examined. PMID- 10366543 TI - Toxic equivalency factors of polychlorinated dibenzo-p-dioxins in an ovulation model: validation of the toxic equivalency concept for one aspect of endocrine disruption. AB - Polychlorinated dibenzo-p-dioxins (PCDDs) are structural analogues, which produce a similar spectrum of biological and toxicological responses in animals, albeit with differential potencies. Very consistent structure-activity relationships have been found for acute toxicity and some biochemical effects among these compounds. For the current experiments, the gonadotropin-primed immature female rat model was used to study the effect of 2,3,7, 8-tetrachlorodibenzo-p-dioxin (TCDD), 1,2,3,7, 8-pentachlorodibenzo-p-dioxin (PeCDD), and 1,2,3,4,7, 8 hexachlorodibenzo-p-dioxin (HxCDD) on ovulation. Single doses of different PCDDs and their mixture were given orally to 23-day-old rats. Gonadotropin from pregnant mare's serum (PMSG) was injected (5 IU) 24 h later to induce follicular maturation. Rats were decapitated at various times after PMSG, blood was collected, and ovarian weight was measured. Serum concentrations of 17beta estradiol (E2), progesterone (P4), luteinizing hormone (LH), follicle stimulating hormone (FSH), and prolactin (PrL) were determined by radioimmunoassay. Ovulation was measured at 72 h after injection of PMSG by counting ova flushed from oviducts. PCDDs dose dependently decreased the number of ova per ovary and reduced ovarian weight gain induced by PMSG. The slopes of the dose-response curves generated by individual PCDDs and/or their mixture were similar. PMSG induced increase in serum E2 was enhanced on the day of expected ovulation by PCDDs; in contrast, serum P4 and FSH were decreased at that same time point. PCDDs also altered the temporal pattern of serum E2, FSH, and LH but not that of PrL. Histologically the effect of all three PCDDs consisted of ova trapped in preovulatory follicles and a lack of or reduced number of corpora lutea. The results indicate that the PCDDs, tested in the present model, have the same mode of action on ovulation and the reproductive hormones, e.g., LH, FSH, P4 and E2. Furthermore, the dose responses of the individual congeners are parallel to each other and also to that of their equipotent mixture, which represent a validation of the TEQ concept for one aspect of endocrine disruption, that is for inhibition of ovulation. PMID- 10366544 TI - N-Nitrosodimethylamine-mediated cytotoxicity in a cell line expressing P450 2E1: evidence for apoptotic cell death. AB - N-Nitrosodimethylamine (NDMA) is an acute hepatotoxin and potent carcinogen. The metabolic activation of NDMA to reactive metabolites is a critical step for the expression of its toxic and carcinogenic potential. We have previously demonstrated a strong correlation between methylation of cellular macromolecules and NDMA-mediated cytotoxicity, and we have demonstrated that reactive oxygen species may partially contribute to the toxic effects in P450 2E1-expressing cells. The mode of cell death in NDMA-treated monolayer cultures exhibited the following characteristics: (i) condensation of nuclear chromatin as demonstrated by using Hoechst 33258 staining, (ii) DNA fragmentation as detected by combining pulsed field and conventional agarose gel electrophoresis, and (iii) DNA double strand breaks determined by using the in situ terminal deoxynucleotidyl transferase assay and flow cytometric analysis. These results indicate that reactive metabolites of NDMA trigger activation of the signal pathway for apoptotic cell death in these P450-expressing cells. The NDMA-mediated cell death was partially prevented by the endonuclease inhibitor, aurintricarboxylic acid, as well as the caspase inhibitors, acetyl-Asp-Glu-Val-Asp-CHO and acetyl-Tyr-Val Ala-Asp-CHO. The cell cycle distribution was altered in NDMA-treated cells resulting in an increase in the G2/M phase and a decrease in the G1 phase. Our results suggest that DNA degradation, the inability to complete DNA repair, the biochemical events associated with G2/M arrest, and the process of apoptotic death all result from P450 2E1-catalyzed metabolism of NDMA. PMID- 10366545 TI - Induction of arylhydrocarbon receptor expression in embryoblast cells of rabbit preimplantation blastocysts upon degeneration of Rauber's polar trophoblast. AB - The arylhydrocarbon receptor (AhR) is a ligand-activated transcription factor and mediates carcinogenic, teratogenic, and toxic effects of xenobiotics such as dioxin and coplanar polychlorinated biphenyls. The AhR nuclear translocator (ARNT) is involved in AhR signal transduction. We have analyzed the expression of AhR and ARNT mRNA and AhR protein in Day 3 pc (postcoitum) rabbit morulae and Days 4 and 6 pc blastocysts using RT-PCR, nested PCR, whole mount in situ hybridization, and whole mount immunohistochemistry with subsequent confocal laser scanning analysis. AhR and ARNT transcripts were detected in all stages investigated, indicating coexpression of both transcription factors. AhR protein was localized in the cytoplasm. It was detected in Day 3 pc morulae and in blastocysts. In Day 4 pc blastocysts, only trophoblast cells but not embryoblast cells were immunopositive. However, at Day 6 pc, the embryoblast cells also expressed AhR protein and this expression was correlated with the degeneration of Rauber's trophoblast layer. PMID- 10366546 TI - Transplacental and lactational transfer of p,p'-DDE in Sprague-Dawley rats. AB - p,p'-DDE (hereafter DDE), a persistent metabolite of p,p'-DDT, is a widespread environmental contaminant that can induce antiandrogenic developmental effects in rats. Quantitative measurements of the transfer of DDE from pregnant or lactating dams to the fetus or suckling neonate were performed, and physiologically based pharmacokinetic (PBPK) models for the transplacental and lactational transfer of DDE were developed. Pregnant Sprague-Dawley rats were dosed by gavage in corn oil with either 10 or 100 mg DDE per kg body wt per day from Gestation Day (gd) 14 to 18. DDE was analyzed in several maternal tissues as well as in fetal and neonatal tissues from gd 15 to Postnatal Day (pnd) 21. Fetal DDE concentrations were about threefold lower than corresponding placental concentrations. By adopting a cross fostering design, the contributions of transplacental and lactational transfer were compared. In the pup liver, where DDE was detectable in the 100 mg/kg groups on pnd 10, the lactationally exposed group had DDE concentrations about 50 times higher than those of the in utero only exposure group; the lactation only exposure groups had DDE tissue dose profiles very similar to those of the in utero plus lactation exposure groups, indicating that the lactational route is far more important than the in utero route quantitatively. The PBPK models postulated initial absorption of DDE into both the blood circulation and lymphatic system with the primary storage sites being maternal and neonatal adipose tissues. Mobilization of DDE from its storage sites is postulated to occur via its association with mobilized fatty acids and lipoproteins. The results provide an overall framework for evaluating the tissue dosimetry of DDE and for understanding how maternal exposure to DDE could affect perinatal sexual development in utero or in the early postnatal period. PMID- 10366547 TI - Biotransformation and toxicokinetics of musk xylene in humans. AB - Musk xylene (1-tert-butyl-3,5-dimethyl-2,4,6-trinitrobenzene, MX) is widely used as a fragrance ingredient in detergents and toiletries and is an environmental contaminant. High concentrations of MX have been found in fish, and humans are constantly exposed to MX as a result of its stability in the environment. We investigated the biotransformation and toxicokinetics of MX in humans. A single dose of 0.3 mg/kg body wt of 15N-labeled MX (15N-MX) was given to six volunteers (three male and three female) by the oral route and to another six volunteers (three males and three females) by the dermal route. Urine was collected for 96 h after exposure. Blood samples were taken at intervals for up to 140 days after administration. The metabolite 1-tert-butyl-3,5-dimethyl-15N-4-amino-2,6 dinitrobenzene in urine and 15N-MX in plasma were quantified by gas chromatography/electron-capture mass spectrometry (GC-MS/NCI). Peak plasma concentrations of 15N-MX after oral administration were 36-262 and 1.6-5.5 ng/ml plasma after dermal administration. The toxicokinetics of 15N-MX in plasma can be described by a two-compartment kinetic model with an initial rapid decrease, due to the distribution from the blood into a second compartment (likely fat tissue) and a terminal elimination phase with an average half-life of 70 days for both routes of administration. The amount of 1-tert-butyl-3,5-dimethyl-15N-4-amino-2,6 dinitrobenzene (15N-4-A-MX) in recovered urine represented 0.1-0.5% of the oral applied dose of 15N-MX, respectively, 0.02-0.16% of dermal dose. After a short time of invasion the concentrations of 15N-4-A-MX in urine reached a maximum 18 24 h after administration. The further elimination of the metabolite occurred by first-order kinetics with an average elimination half-life of 11.8 h. After the single oral or dermal dose of 15N-MX, 15N-4-A-MX was not detected in hemoglobin. However, hemoglobin samples contained 1-tert-butyl-3, 5-dimethyl-4-amino-2,6 dinitrobenzene (4-A-MX) (11.4-18.9 fmol/mg Hb), likely derived from chronic environmental exposures. PMID- 10366548 TI - Effects of immigration on the dynamics of simple population models. AB - Many simple population models exhibit the period doubling route to chaos as a single parameter, commonly the growth rate, is increased. Here we examine the effect of an immigration process on such models and explain why in the case of one-dimensional ("single-humped") maps, immigration often tends to suppress chaos and stabilise equilibrium behaviour or cyclical oscillations of long period. The conditions for which an increase of immigration "simplifies" population dynamics are examined. PMID- 10366549 TI - Some statistical improvements for estimating population size and mutation rate from segregating sites in DNA sequences. AB - In population genetics, under a neutral Wright-Fisher model, the scaling parameter straight theta=4Nmu represents twice the average number of new mutants per generation. The effective population size is N and mu is the mutation rate per sequence per generation. Watterson proposed a consistent estimator of this parameter based on the number of segregating sites in a sample of nucleotide sequences. We study the distribution of the Watterson estimator. Enlarging the size of the sample, we asymptotically set a Central Limit Theorem for the Watterson estimator. This exhibits asymptotic normality with a slow rate of convergence. We then prove the asymptotic efficiency of this estimator. In the second part, we illustrate the slow rate of convergence found in the Central Limit Theorem. To this end, by studying the confidence intervals, we show that the asymptotic Gaussian distribution is not a good approximation for the Watterson estimator. PMID- 10366550 TI - Recombination as a point process along sequences. AB - Histories of sequences in the coalescent model with recombination can be simulated using an algorithm that takes as input a sample of extant sequences. The algorithm traces the history of the sequences going back in time, encountering recombinations and coalescence (duplications) until the ancestral material is located on one sequence for homologous positions in the present sequences. Here an alternative algorithm is formulated not as going back in time and operating on sequences, but by moving spatially along the sequences, updating the history of the sequences as recombination points are encountered. This algorithm focuses on spatial aspects of the coalescent with recombination rather than on temporal aspects as is the case of familiar algorithms. Mathematical results related to spatial aspects of the coalescent with recombination are derived. PMID- 10366551 TI - The coalescent time in the presence of background fertility selection. AB - Selection ultimately entails differential reproductive success over several generations. This can be measured as a correlation of the number of progeny an individual has with the number of progeny its parent had. This correlation could have a genetic or a cultural basis. The effect of such a correlation is to multiply the single generation sampling variance (Vdeltap) in the diffusion approximation for fixation time by (1-b)+bx(1+r)/(1-r), where bxrn is the correlation between the number of progeny of an individual and its ancestor n generations ago (e.g., b is the heritability and br is the resultant parent offspring progeny number correlation if the progeny number is genetically determined). This results in a reduction of the fixation or coalescent time by division by this factor. Sex differences in this correlation have been observed, and this provides an explanation for the difference of coalescent times of y chromosomes and mitochondria. PMID- 10366552 TI - Local frequency dependence and global coexistence. AB - In sessile organisms such as plants, interactions occur locally so that important ecological aspects like frequency dependence are manifest within local neighborhoods. Using probabilistic cellular automata models, we investigated how local frequency-dependent competition influenced whether two species could coexist. Individuals of the two species were randomly placed on a grid and allowed to interact according to local frequency-dependent rules. For four different frequency-dependent scenarios, the results indicated that over a broad parameter range the two species could coexist. Comparisons between explicit spatial simulations and the mean-field approximation indicate that coexistence occurs over a broader region in the explicit spatial simulation. PMID- 10366553 TI - The effects of density dependence and immigration on local adaptation and niche evolution in a black-hole sink environment. AB - We examine the effects of density dependence and immigration on local adaptation in a "black-hole sink" habitat, i.e., a habitat in which isolated populations of a species would tend to extinction but where a population is demographically maintained by recurrent one-way migration from a separate source habitat in which the species persists. Using a diploid, one-locus model of a discrete-generation sink population maintained by immigration from a fixed source population, we show that a locally favored allele will spread when rare in the sink if the absolute fitness (or, in some cases, the geometric-mean absolute fitness) of heterozygotes with the favored allele is above one in the sink habitat. With density dependence, the criterion for spread can depend on the rate of immigration, because immigration affects local densities and, hence, absolute fitness. Given the successful establishment of a locally favored allele, it will be maintained by a migration-selection balance and the resulting polymorphic population will be sustained deterministically with either stable or unstable dynamics. The densities of stable polymorphic populations tend to exceed densities that would be maintained in the absence of the favored allele. With strong density regulation, spread of the favored allele may destabilize population dynamics. Our analyses show that polymorphic populations which form subsequent to the establishment of favorable alleles have the capacity to persist deterministically without immigration. Finally, we examined the probabilistic rate at which new favored alleles arise and become established in a sink population. Our results suggest that favored alleles are established most readily at intermediate levels of immigration. PMID- 10366554 TI - Genetic differentiation in populations with different classes of individuals. AB - I formulate and analyse a model of population structure with different classes of individuals. These different classes may be age classes, other demographic classes, or different types of habitats homogeneously distributed over a geographical area. The value of population differentiation under an island model of dispersal and the increase of differentiation with geographical distance in one- and two-dimensional "isolation by distance" models are then obtained for a generalization of the FST measure of population structure, as a function of "effective" mutation, migration, and population size parameters. The relevant effective subpopulation size is related to the "mutation effective population size" of a single isolated subpopulation and, in models of age-structured populations, to the inbreeding effective population size. PMID- 10366555 TI - Evolution of mutualistic symbiosis without vertical transmission. AB - Mutualistic symbioses are considered to evolve from parasitic relationships. Vertical transmission, defined as the direct transfer of infection from a parent organism to its progeny, has been suggested as a key factor causing reduction of symbiont virulence and evolution of mutualism. On the other hand, there are several mutualistic associations without vertical transmission, such as those between plants and mycorrhizal fungi, legumes and rhizobia, and some corals and dinoflagellates. It is expected that all mutualisms evolve perfect vertical transmission if the relationship is really mutualistic, because hosts may fail to acquire symbionts if they do not transmit the symbionts by vertical transmission. We have developed a mathematical model to clarify the conditions under which mutualistic symbiosis without vertical transmission should evolve. The evolution may occur when and only when (i) vertical transmission involves some costs in the host, (ii) the symbiont suffers direct negative effects if it exploits the host too intensively, (iii) the host establishes the ability to make use of waste products from the symbiont, and (iv) the mechanism of vertical transmission is controlled by the host. We also clarify the conditions under which mutualistic symbiosis with vertical transmission evolves. PMID- 10366556 TI - Evolutionary dynamics of seed size and seedling competitive ability. AB - We present a model for the evolutionary dynamics of seed size when there is a trade-off between seed size and seed number, and seedlings from large seeds are better competitors and have a higher precompetitive survival than seedlings from small seeds. We find that strong competitive asymmetry, high resource levels, and intermediate harshness of the precompetitive environment favor coexistence of plants with different seed sizes. If the evolution of seed size is mutation limited and single mutations have only a small phenotypic effect, then an initially monomorphic population reaches the final evolutionarily stable polymorphic state through one or more discrete evolutionary branching events. At each such branching event, a given lineage already present in the population divides into two phenotypically diverging daughter lines, each with its own seed size. If the precompetitive survival of seeds and seedlings is high for small and large seeds alike, however, evolutionary branching may be followed by the extinction of one or more lineages. Various results presented here are model independent and point the way to a more general evolutionary bifurcation theory describing how the number and stability properties of evolutionary equilibria may change as a consequence of changes in model parameters. PMID- 10366557 TI - The dileucine-based sorting motif in HIV-1 Nef is not required for down regulation of class I MHC. AB - A dileucine-based protein sorting motif has recently been identified within the C terminal, solvent-exposed loop of HIV-1 Nef and has been shown to be required for Nef-mediated down-regulation of CD4 and for optimal viral infectivity. Here, we report that mutation of the dileucine motif has no effect on Nef-mediated down regulation of class I MHC heavy chain. Instead, deletion of an acidic domain just N-terminal of the polyproline helix of the SH3-binding domain significantly impairs this function. These data indicate that down-regulation of class I MHC and CD4 are mechanistically distinct processes. The data also suggest that protein interactions mediated by the acidic domain, rather than by the dileucine motif, may contribute to this function of Nef. PMID- 10366558 TI - Specific and nonspecific immune stimulation of MHC-II-deficient mice results in chronic HSV-1 infection of the trigeminal ganglia following ocular challenge. AB - Ocular herpes simplex virus type 1 (HSV-1) infection of MHC-II-deficient mice (AO/Obeta mice) or their parental C57BL/6J wild-type mice resulted in the establishment of typical HSV-1 latent infections in the trigeminal ganglia (TG) of the surviving mice by day 28 postinfection. Latency was characterized by the complete absence of infectious virus in TG extracts, the ability to recover latent virus only following prolonged tissue culture cultivation of explanted TG, and the presence of HSV-1 DNA in TG extracts. When mice were vaccinated prior to ocular HSV-1 challenge, latency appeared unaltered in the C57BL/6J wild-type mice. However, in AO/Obeta mice, clearance of virus from the TG appeared to be seriously impaired, resulting in a chronic productive infection, rather than a latent infection. Infectious virus was readily detected in TG extracts of vaccinated AO/Obeta mice until at least 63 days postinfection. Glycoprotein B mRNA was also readily detected, confirming continued viral transcription. These chronic infections occurred regardless of whether the AO/Obeta mice were vaccinated with HSV-1-specific antigens (i.e., live HSV-1 strain KOS, recombinantly expressed HSV-1 glycoprotein D plus Freund's adjuvant, or a mixture of seven recombinantly expressed HSV-1 glycoproteins plus adjuvant) or non-HSV-1 specific antigens (i.e., tissue culture medium plus 5% fetal bovine serum, the expression vector plus adjuvant, or adjuvant alone). Passive transfer of HSV-1 neutralizing antibody to vaccinated AO/Obeta mice between days 0 and 28 post ocular challenge did not clear infectious virus from the TG. Passive transfer of anti-HSV-1 antibody or purified naive mouse serum to unvaccinated AO/Obeta mice on days 3 or 6 post-HSV-1 ocular challenge also resulted in chronic, rather than latent, infection of the TG. Passive transfer of naive sera from B-cell-deficient mice or injection of keyhole limpet hemocyanin or purified IgG, but not PBS or dextran, 3 days after HSV-1 challenge also resulted in chronic infection of the TG. PMID- 10366559 TI - Both RNA rearrangement and point mutation contribute to repair of defective chimeric viral genomes to form functional hybrid viruses in plants. AB - The putative movement protein gene (p27) plus 5' and 3' flanking sequences of cucumber leaf spot aureusvirus (CLSV) was inserted into an infectious cucumber necrosis tombusvirus (CNV) cDNA clone containing a deletion in the cell-to-cell movement protein gene. Approximately 5% of plants inoculated with synthetic transcripts of two such defective chimeric CNV/CLSV cDNA clones developed systemic symptoms 7-19 days postinoculation. Reverse transcription-polymerase chain reaction and sequence analysis of virus obtained from systemically infected leaves indicated that both point mutation and RNA rearrangement (deletion) contributed to the formation of movement competent CNV/CLSV hybrid viruses. The hybrid viruses were found to accumulate to high levels in infected plants, to form stable virions, and to be mechanically transmissible. In addition, a hybrid virus that lacked 50 amino acids at the carboxyl-terminal region of CLSV p27 was still capable of facilitating CNV movement. These data provide experimental evidence for the role of CLSV p27 in viral cell-to-cell movement and demonstrate that p27 can enable efficient movement of the CNV genome. Moreover, the data show that RNA rearrangements known to occur during CNV RNA replication can contribute to rapid evolution of the CNV genome. PMID- 10366560 TI - Effects of egg-adaptation on the receptor-binding properties of human influenza A and B viruses. AB - Propagation of human influenza viruses in embryonated chicken eggs (CE) results in the selection of variants with amino acid substitutions near the receptor binding site of the hemagglutinin (HA) molecule. To evaluate the mechanisms by which these substitutions enable human virus growth in CE, we studied the binding of 10 human influenza A (H1N1, H3N2) and B strains, isolated and propagated solely in MDCK cells, and of their egg-adapted counterparts to preparations of cellular membranes, gangliosides, sialylglycoproteins, and sialyloligosaccharides. All egg-adapted variants differed from nonadapted strains by increased binding to the plasma membranes of chorio-allantoic (CAM) cells of CE and by the ability to bind to CAM gangliosides. In addition, there was no decrease in affinity for inhibitors within allantoic fluid. These findings indicate that growth of human influenza viruses in CE is restricted because of their inefficient binding to receptors on CAM cells and that gangliosides can play an important role in virus binding and/or penetration. The effects of the egg-adaptation substitutions on the receptor-binding properties of the viruses include (i) enhancement of virus binding to the terminal Sia(alpha2-3)Gal determinant (substitutions in HA positions 190, 225 of H1N1 strains and in position 186 of H3N2 strains); (ii) a decrease of steric interference with more distant parts of the Sia(alpha2-3Gal)-containing receptors (a loss of glycosylation sites in positions 163 of H1 HA and 187 of type B HA); and (iii) enhanced ionic interactions with the negatively charged molecules due to charged substitutions at the tip of the HA [187, 189, 190 (H1), and 145, 156 (H3)]. Concomitantly with enhanced binding to Sia(alpha2-3)Gal-terminated receptors, all egg-adapted variants decreased their affinity for equine macroglobulin, a glycoprotein bearing terminal 6'-sialyl(N-acetyllactosamine)-moieties. PMID- 10366561 TI - A 189-bp repeat region within the human cytomegalovirus replication origin contains a sequence dispensable but irreplaceable with other sequences. AB - The human cytomegalovirus (HCMV) replication origin exhibits a strain-dependent difference in the number of copies of a 189-bp region: the AD169 and Towne strains contain one and three copies of the region, respectively. A nearly complete deletion of the 189-bp repeat region of the Towne strain does not eliminate the origin's ability to initiate DNA synthesis. Here we report that the replication ability of the HCMV replication origin in infected cells disappeared after replacements of an internal sequence (152 bp) of the 189-bp repeat region with lambda DNA of identical and different lengths as well as after introduction of multiple nucleotide substitutions within the 152-bp internal sequence of the 189-bp repeat. In contrast, a variation in the copy number of 189-bp region (either one or two copies) or an inversion of the 152-bp internal sequence of the 189-bp repeat maintained replication abilities similar to those of the wild-type origin of the Towne strain. These results indicate that the 189-bp repeat region within the HCMV replication origin is not just a dispensable spacer sequence but instead contains an irreplaceable sequence that may play a supporting role in HCMV DNA replication. PMID- 10366562 TI - Internal initiation of translation of bovine viral diarrhea virus RNA. AB - Initiation of translation on the bovine viral diarrhea virus (BVDV) internal ribosomal entry site (IRES) was reconstituted in vitro from purified translation components to the stage of 48S ribosomal initiation complex formation. Ribosomal binding and positioning on this mRNA to form a 48S complex did not require the initiation factors eIF4A, eIF4B, or eIF4F, and translation of this mRNA was resistant to inhibition by a trans-dominant eIF4A mutant that inhibited cap mediated initiation of translation. The BVDV IRES contains elements that are bound independently by ribosomal 40S subunits and by eukaryotic initiation factor (eIF) 3, as well as determinants that mediate direct attachment of 43S ribosomal complexes to the initiation codon. PMID- 10366563 TI - Herpes simplex virus induces intracellular redistribution of E2F4 and accumulation of E2F pocket protein complexes. AB - Accumulation of E2F-p107 and E2F-pRB DNA binding complexes occurred after herpes simplex virus infection of U2-OS cells. Accumulation of E2F-p107 also occurred by 4 h p.i. in C33 cells. This corresponded to a time when host DNA synthesis was reduced by 50%, and lagged by >/=1 h, the onset of viral DNA synthesis. To determine the basis for increased nuclear E2F complexes, we investigated the effects of virus infection on the intracellular distribution of the E2F-dependent DNA binding complexes and their protein constituents. Western blot analyses of whole cell extracts revealed that amounts of E2F4, E2F1, DP1, and p107 remained unchanged after infection of C33 cells. Analysis of cytoplasmic and nuclear fractions, however, revealed that cytoplasmic E2F4 decreased and nuclear E2F4 increased. This correlated with a loss of cytoplasmic E2F DNA-binding activity and a corresponding increase in nuclear DNA-binding activity. Concomitant with its redistribution, the apparent molecular weight of total and p107-associated E2F4 increased, at least partially as a result of protein phosphorylation. Increased nuclear E2F-pRB in U2-OS cells was accompanied by the conversion of pRB from a hyper- to a hypophosphorylated state. Infection of U2-OS cells with viral mutants indicated that viral protein IE ICP4 was necessary for the decrease in cytoplasmic E2F-p107, and that viral protein DE ICP8 was required for nuclear accumulation of p107-E2F. In contrast, ICP8 was not required for accumulation of E2F-pRB. These results indicate that the increase in E2F-p107 may be explained by the redistribution and modification of E2F4 and the increase in E2F-pRB by modification of pRB. PMID- 10366565 TI - The role of gamma interferon in immune resistance to vaginal infection by herpes simplex virus type 2 in mice. AB - We investigated the role of interferon gamma (IFN-gamma) in a mouse model of immunity to vaginal infection by herpes simplex virus type 2 (HSV-2). Within 8 h after immune mice were challenged intravaginally with HSV-2, IFN-gamma concentrations in vaginal secretions reached levels that can be antiviral in vitro. This rapid synthesis of IFN-gamma occurred in immune-challenged mice but not in nonimmune-challenged mice, indicating that it required memory T cells. Immunostaining and in situ hybridization revealed that the IFN-gamma was synthesized by cells whose morphological appearance suggested that they were lymphocytes and macrophage-like cells in the mucosa. The presence of IFN-gamma in vaginal secretions was correlated with upregulation of MHC class II antigens in the epithelium and with vigorous (30-fold) recruitment of T and B lymphocytes into the vagina. In vivo administration of anti-IFN-gamma to immune mice 17 h before virus challenge blocked the subsequent appearance of IFN-gamma in vaginal secretions, blocked upregulation of class II antigens, blocked adherence of T cells to endothelium and their recruitment into the vagina, and markedly reduced immunity against reinfection of the vaginal epithelium. PMID- 10366564 TI - Activation of Ste20 by Nef from human immunodeficiency virus induces cytoskeletal rearrangements and downstream effector functions in Saccharomyces cerevisiae. AB - The negative factor (Nef) from human and simian immunodeficiency viruses is important for the pathogenesis of acquired immune deficiency syndrome. Among other targets, it activates the Nef-associated kinase, which is related to the p21-activated kinase. In this study, we demonstrate that Nef activates Ste20, the homolog of p21-activated kinase in Saccharomyces cerevisiae. Nef binds to the adaptor proteins Bem1 and Ste20 via its proline-rich (PXXP) and diarginine (RR) motifs, respectively. These interactions induce the mitogen-activated protein kinase and increase the rates of budding, sizes of cells, and patterns of mating projections. These effects of Nef depend on the small GTPase Cdc42 and guanine nucleotide exchange factor Cdc24. Thus, studies in S. cerevisiae identified specific interactions between Nef and cellular proteins and their associated signaling cascade. PMID- 10366566 TI - Tetracycline-mediated regulation of gene expression within the human cytomegalovirus genome. AB - To evaluate the utility of tetracycline gene regulation in the study of human cytomegalovirus gene functions, expression of luciferase under the control of tetracycline-regulatable promoters was studied following transient plasmid transfections and from within recombinant human cytomegalovirus genomes. The tetracycline-regulatable promoter PhCMV*-1 contains sequences from the human cytomegalovirus ie1/ie2 promoter and seven upstream tet operator sites which bind the activator protein tTA only in the absence of tetracycline (Gossen and Bujard (1992). Proc. Natl. Acad. Sci. USA 89, 5547-5551). Two modifications of PhCMV*-1 were also studied: P1129, in which the tet operator sites were reduced from seven to one; and P1125, in which human cytomegalovirus sequences were replaced by adenovirus major late promoter and terminal deoxynucleotidyltransferase initiator sequences. In transient assays, PhCMV*-1 and P1125 exhibited modest differential regulation but were strongly activated by viral infection. P1129 exhibited less viral activation and narrower regulation. In the viral genome, PhCMV*-1 exhibited regulation up to 7-fold during late times of infection, whereas P1125 displayed nearly 100-fold regulation. Regulation of P1125 was fully reversed within 12 to 24 h of adding or removing tetracycline. These results suggest that P1125 may provide sufficient conditional expression to effectively regulate human cytomegalovirus late genes. PMID- 10366567 TI - Transcript mapping and transregulatory behavior of varicella-zoster virus gene 21, a latency-associated gene. AB - Gene 21 is one of at least four genes transcribed during latent infection of varicella-zoster virus (VZV) in human ganglia. It may encode a nucleocapsid protein, but its function in lytic and latent infection is not clear. To characterize further the structure and the function of the gene 21 open reading frame (ORF 21), precise localization of its transcripts and their termini was determined by using Northern analysis, S1 nuclease or RNase protection, and primer extension assays. One abundant 3.5-kb transcript that spans ORF 21 was identified. A predominant transcription start site was defined at -78 nucleotide (nt) relative to the ORF 21 translation start codon ATG, and two potential TATA elements were identified at 26 and 83 nt upstream of the 5' end of gene 21 transcripts. Transcription was found to terminate 210 nt beyond the ORF 21 translation stop codon and immediately before the start codon of ORF 22. In transient expression assays, the ORF 21 showed no significant transregulatory activity on promoters of diverse kinetic classes. The ORF 21 promoter, however, was transactivated strongly by VZV infection or by ORF 62. PMID- 10366568 TI - Use of site-specific mutagenesis and monoclonal antibodies to map regions of CD46 that interact with measles virus H protein. AB - Researchers at our laboratory have been dissecting the binding domains of the receptor for the Edmonston laboratory strain of measles virus (CD46) through site specific mutagenesis. We initially substituted most of the hydrophilic amino acids in the two external short consensus regions (SCRI and SCRII) of CD46 with the amino acid alanine [Hsu et al. (1997) J. Virol. 71:6144-6154] and found that the glutamic-arginine residues at positions 58 and 59 were particularly sensitive to change. Here we consider the roles of hydrophobic amino acids in the binding between measles virus H protein and CD46. Hydrophobic amino acids in the SCRI and SCRII domains of CD46 were systematically replaced with serine. The effects of these changes were monitored through the interaction of Sf9 insect cells expressing the H protein and mouse OST-7 cells synthesizing the mutant CD46 molecules. Binding was quantified through a colorimetric assay for beta galactosidase that was also produced by the insect cells. Our results indicate that E45, Y54, 58E/R59, Y68, F69, Y101, I102, R103, D104, and Y117 seem to be critical residues for the binding of CD46 to measles virus H protein. The hydrophilic amino acid R59 in SCR1 and hydrophobic residues Y101, I102, and Y117 in SCR2 seem to be especially important for interaction between H protein and CD46. In addition, we mapped the antigenic epitopes of five monoclonal antibodies that are known to inhibit the binding between H protein and CD46. Three of these antibodies recognized regions in SCR1, and two reacted with amino acids in SCR2. For the most part, the determinants recognized by the monoclonal antibody corresponded to the amino acids that were most sensitive to change in the binding process. The SCR1 and SCR2 domains of CD46 were modeled from an analogous region in another complement regulatory protein, factor H, whose three-dimensional structure has been previously reported. Amino acids implicated in binding seem to lie on one planar face of the SCR1 and SCR2 domains. These studies serve as a prelude to understanding the structural interactions that occur between CD46 and the measles virus H protein. PMID- 10366569 TI - Nuclear localization of human immunodeficiency virus type 1 integrase expressed as a fusion protein with green fluorescent protein. AB - Lentiviruses in general and the human immunodeficiency virus type 1 (HIV-1) in particular have the ability to integrate their genome stably into the chromosome of nondividing cells. Integration of HIV cDNA is mediated by the viral integrase (IN). Apart from its catalytic activity, this enzyme seems to play an important role in the transport of the HIV preintegration complex into the nucleus of nondividing cells. We studied the karyophilic properties of IN by constructing an N-terminal fusion protein of HIV-1 integrase and green fluorescent protein (GFP IN). Transient expression of GFP-IN in various mammalian cell lines was demonstrated by fluorescence microscopy, flow cytometry, and Western blotting. Although wild-type GFP was localized throughout the cell, GFP-IN was localized predominantly in the nucleus. Nuclear localization of GFP-IN was also obtained after transient transfection of the cells arrested in the G1/S phase of the cell cycle. These results provide compelling evidence for the karyophilic properties of the HIV-1 integrase. PMID- 10366570 TI - Sequence comparison of JSRV with endogenous proviruses: envelope genotypes and a novel ORF with similarity to a G-protein-coupled receptor. AB - Ovine pulmonary carcinoma, a contagious lung cancer of sheep, is caused by the oncogenic jaagsiekte sheep retrovirus (JSRV) that is closely related to a family of endogenous sheep retroviral sequences (ESRVs). By using exogenous virus specific U3 oligonucleotide primers, the entire JSRV proviral genome or its 3' part was amplified from tumor DNA. Analysis of these proviral sequences revealed a novel open reading frame (ORF) within the pol coding region, designated ORF X, which was well conserved in ESRV and JSRV sequences. Deduced amino acids of ORF X showed similarity to a portion of the mammalian adenosine receptor subtype 3, a member of the G-protein-coupled receptor family. Comparison of deduced env amino acids of six JSRV strains from three continents identified 15 residues that defined two distinct genotypes of JSRVs. Sequence analysis identified two highly variable regions between JSRV and ESRV in the transmembrane domain of env (TM) and the 3' unique sequence (U3) of the long terminal repeat, from which JSRV specific DNA probes were derived. By using these DNA probes in Southern hybridization, for the first time we successfully identified JSRV proviral sequences in tumor genomic DNA in the presence of multiple ESRV loci, validating the use of exogenous virus-specific DNA probes in the analysis of oncogenic proviral integration sites and identification of integrated exogenous proviral sequences. PMID- 10366571 TI - Response to IL-6 of HPV-18 cervical carcinoma cell lines. AB - The human papillomavirus type 18 (HPV-18) upstream regulatory region (URR) controls cell type-specific expression of the viral oncoproteins E6 and E7. The HPV-18 URR is active in the cervical carcinoma cell line HeLa but inactive in the hepatoma cell line HepG2. C/EBPss (NF-IL-6) was shown to participate as an important regulator in HPV transcription dependent on the cell type. The finding that C/EPBss is critical for HPV-18 URR activity and that C/EPBss is induced by IL-6 offers the opportunity of manipulating HPV activity by specific cytokine treatment. In this report, we show that treatment with IL-6 results in activation of HPV-18 URR activity in HepG2 cells. In contrast, the HPV-18 URR is not inducible by IL-6 in three cervical carcinoma cell lines. In all three cell lines we found decreased expression of the IL-6 receptor compared to the IL-6 responsive HepG2 cells, whereas the level of expression of the signal transduction component gp130 is present in all cells. These results suggest that cervical carcinoma cells may circumvent the IL-6-induced cellular defense mechanism through downregulation of the IL-6-receptor. PMID- 10366572 TI - CD4(+) T-cells are required for the establishment of maedi-visna virus infection in macrophages but not dendritic cells in vivo. AB - The role of CD4(+) lymphocytes in the establishment of lentivirus infection in macrophages has been studied in an in vivo system of lentivirus infection where CD4(+) lymphocytes are not the targets for infection. Using the non-T-cell-tropic lentivirus, maedi-visna virus (MVV), in CD4-depleted sheep, we have found that CD4(+) T cells were required for MVV infection in macrophages but not dendritic cells. CD4-depleted sheep had significantly lower levels of MVV-infected cells in lymph nodes and efferent lymph after MVV challenge in the drainage area of the lymph node. Due to the absence of virus in combination with the lack of CD4(+) T helper cells, virus-specific immune responses were reduced. There was delayed induction of cytotoxic T cell precursors, a marked reduction in virus-specific in vitro proliferative responses, and a delay in the appearance of MVV-specific antibodies. By contrast, CD4 depletion had no effect on the establishment of MVV infection in afferent lymph dendritic cells migrating from the skin infection site to the lymph node. PMID- 10366573 TI - Functional characterization of the HveA homolog specified by African green monkey kidney cells with a herpes simplex virus expressing the green fluorescence protein. AB - We cloned the gene specified by African monkey kidney cells (Vero) that codes for the homolog of the herpes virus entry mediator (HveA) specified by HeLa cells. The primary sequence of the monkey HveA (HveAs) differed significantly from HveA. Single amino acid differences were distributed throughout the amino and carboxyl terminal portions of the HveAs in comparison with the HveA, whereas certain regions were highly conserved. The predicted membrane spanning domains of the two receptors differed substantially due to insertions and deletions of short amino acid sequences. The ability of HveAs to mediate HSV virus entry was tested in a series of experiments using the recombinant virus KOS/EGFP, which constitutively expressed the enhanced green fluorescence protein (EGFP) and Chinese hamster ovary cells (CHO) transformed with the HveAs gene. The KOS/EGFP virus was constructed by inserting an EGFP gene cassette within the intergenic region between the UL53 (gK) and UL54 (ICP27) genes. The KOS/EGFP virus formed viral plaques and replicated as well as the wild-type KOS virus. HveAs-transformed CHO cells constitutively expressing HveAs mediated herpesvirus entry efficiently, whereas cells transformed with the HveAs gene in the noncoding orientation did not mediate virus entry. A genetically engineered protein composed of the amino terminal portion of the HveAs protein fused to the heavy chain of mouse IgG immunoglobulin as well as mouse antibodies raised against HveAs blocked virus entry into HveAs-transformed CHO cells. Thus, HveAs is the functional homolog of HveA. PMID- 10366574 TI - Nitric oxide down-regulates Epstein-Barr virus reactivation in epithelial cell lines. AB - Nitric oxide (NO), a mediator of biological functions, has an antimicrobial activity against a variety of pathogens including viruses. In this study, we found that a constitutive, low level of inducible NO synthase (iNOS) mRNA was expressed in the EBV-infected gastric tissue-derived GT38 and GT39 cell lines, by analysis with the reverse transcription-polymerase chain reaction (RT-PCR) and Southern blotting. Treatment of these cells with a specific NOS inhibitor, NG monomethyl-L-arginine (L-NMMA), induced the immediate-early, EBV transactivator gene BZLF1 protein ZEBRA, suggesting a significant increase in EBV reactivation by L-NMMA. Northern blotting demonstrated that BZLF1 and BRLF1 transcripts were also induced by 12-O-tetradecanoylphorbol-13 acetate (TPA). Meanwhile, constitutive expression of iNOS mRNA was inhibited by TPA. L-NMMA also enhanced TPA-induced expression of the BZLF1 gene. On the other hand, a NO donor, S nitroso-N-acetylpenicillamine (SNAP), which releases NO in an aqueous solution, inhibited the TPA-induced BZLF1 gene expression in a dose-dependent manner at both mRNA and protein levels. These results demonstrated that NO is a regulatory factor in maintaining virus latency via inhibiting EBV reactivation in the infected epithelial cells. PMID- 10366575 TI - Neutralizing antibody responses and evolution of antigenic variants in monozygotic twin lambs infected with phenotypically distinct ovine lentiviruses. AB - Ovine lentivirus (OvLV) isolates 85/34 (OvLV 34) and 84/28 (OvLV 28) were initially characterized as phenotypically distinct "rapid/high" and "slow/low" strains based on replication kinetics, syncytiogenesis, and cell lysis in vitro. In the present study, sera from OvLV-34- or OvLV-28-infected monozygotic twin lambs defined these virus strains as distinct neutralization serotypes. We also show that immune recognition of at least one OvLV neutralization epitope is influenced by genetic differences between lambs. Additional studies determined the neutralization phenotype of virus isolates from alveolar macrophages of OvLV 34- or OvLV-28-infected lambs, evaluated the role of neutralizing antibodies in selection and persistence of antigenic variants, and related the severity of OvLV induced lymphoid interstitial pneumonia (LIP) to the evolution of neutralization variants. These studies demonstrate that (i) macrophage-associated OvLV neutralization variants can arise in the presence or the absence of neutralizing antibodies directed to inoculum viruses, (ii) OvLV variants persist in macrophages in the presence of serum neutralizing antibodies, and (iii) the emergence of OvLV variants is apparently unrelated to the severity of LIP. PMID- 10366576 TI - Neuronal expression of NOS-1 is required for host recovery from viral encephalitis. AB - The role of nitric oxide synthase (NOS) in host defense and clearance of vesicular stomatitis virus (VSV) from the central nervous system (CNS) was examined. NOS-1, NOS-2, and NOS-3 knockout mice were infected with VSV and were treated with either IL-12 or medium. IL-12 treatment resulted in substantially decreased VSV titers in wildtype and NOS-3 knockout mice, but had a marginal effect in the NOS-1 and NOS-2 knockout mice. NOS-1 expression in neurons was associated with survival from VSV infection. The data indicate that the enzyme activity is local, since NOS-2 expression in microglia and inflammatory macrophages and NOS-3 expression in astrocytes, endothelial cells, and ependymal cells did not compensate. PMID- 10366577 TI - Broadly cross-reactive, high-affinity antibody to hypervariable region 1 of the hepatitis C virus in rabbits. AB - The HCV hypervariable region 1 (HVR1) of the main E2 envelope protein is critically important in HCV neutralization but its extreme variability makes immune therapy and vaccine development particularly difficult. To explore the hypothesis that HVR1 carries a common epitope susceptible of eliciting cross reactive neutralizing and inhibitory antibodies, rabbits were immunized with a series of synthetic HVR1 peptides. The anti-HVR1 produced were purified and characterized. Several lines of evidence supported the working hypothesis: (1) although injected only once, a boosting effect from poorly homologous peptides was observed; (2) purified rabbit IgG reacted with high affinity with immunizing peptides and cross-reacted with 16 of 17 unrelated HVR1 peptides; (3) antibodies appeared of restricted diversity irrespective of the linear HVR1 peptide sequences; (4) anti-HVR1 peptides effectively captured HCV in 22 of 33 plasmas from random infected patients; (5) anti-HVR1 IgG blocked the binding of antibody captured HCV to MOLT-4 cells. These findings suggest that with an appropriate HVR1 peptide immunization scheme, high titer, broadly cross-reactive, blocking antibodies to HCV can be produced. Antibodies to the putative ubiquitous HVR1 epitope may have important clinical uses. PMID- 10366578 TI - Expression of the HPV E7 oncoprotein mimics but does not evoke a p53-dependent cellular DNA damage response pathway. AB - Acute expression of the human papillomavirus E7 oncoprotein in preimmortal human fibroblasts induces changes in the abundances of multiple cellular regulatory proteins. These alterations include a destabilization of the retinoblastoma tumor suppressor protein pRB, stabilization of the tumor suppressor protein p53, and increases in the level of the cyclin-dependent kinase inhibitor p21(cip1). Since the HPV E7 oncoproteins can interfere with several cell cycle checkpoints and similar alterations in the levels of pRB, p53, and p21(cip1) are also observed in a p53-dependent response to DNA damage, we investigated whether E7 expression triggers this signal transduction pathway. The results demonstrate that E7 mediated destabilization of pRB does not require p53 activity and is independent of the ability of E7 to induce apoptosis. Moreover, E7-mediated increases in p21(cip1) levels are largely p53-independent and involve stabilization of the p21(cip1) protein. In contrast the decreases in pRB expression in response to DNA damage involve transcriptional downregulation of RB gene expression. PMID- 10366579 TI - Domains of Rinderpest virus phosphoprotein involved in interaction with itself and the nucleocapsid protein. AB - The yeast two-hybrid system was used to identify domains involved in specific in vivo interactions between the Rinderpest virus (RPV) phosphoprotein (P) and nucleocapsid protein (N). N and P genes were cloned in both the yeast GAL4 DNA binding and GAL4 activation domain vectors, which enabled analysis of self and interprotein interactions. Mapping of the domain of P protein involved in its association with itself revealed that the COOH-terminal 32 amino acids (316-347) that forms a part of the highly conserved coiled coil region is important for interaction. In addition, just the coiled coil region of RPV P protein fused to the DNA-binding domain and activation domain of GAL4 was found to be sufficient to bring about activation of the beta-galactosidase reporter. Similarly, mapping of the domains of P protein involved in its interaction with N protein revealed that NH2-terminal 59 amino acids and COOH-terminal 32 amino acids (316-347) involved in P-P interaction are simultaneously required for association with N protein. Interestingly, a P protein mutant with just the NH2-terminal 59 amino acids and the coiled coil domain with all other P protein regions deleted retained its ability to interact with N protein. Furthermore, we were able to show N and P protein interaction in vitro using recombinant N and P proteins expressed in Escherichia coli, demonstrating the existence of direct physical interaction between the two proteins. PMID- 10366580 TI - Isolation and molecular characterization of a novel cytopathogenic paramyxovirus from tree shrews. AB - A cytopathic infectious agent was isolated from the kidneys of an apparently healthy tree shrew (Tupaia belangeri) that had been captured in the area around Bangkok. The infectivity was propagated in Tupaia fibroblast and kidney cell cultures. Paramyxovirus-like pleomorphic enveloped particles and helical nucleocapsids were observed by electron microscopy and accordingly the infectious agent was termed Tupaia paramyxovirus (TPMV). However, no serological cross reactions were detected between TPMV and known paramyxoviruses. For the molecular characterization of TPMV an experimental strategy that allows the random-primed synthesis of relatively large cDNA molecules from viral genomic RNA was applied. Nucleotide sequence analysis of a TPMV-specific cDNA fragment (1544 bp) revealed two nonoverlapping partial open reading frames corresponding to paramyxoviral N and P transcription units. Using modified rapid amplification of cDNA ends techniques, a substantial contiguous portion of the viral genome (4065 nt) was elucidated including the complete N and P/V/C genes. The coding strategy of TPMV as well as significant amino acid sequence homologies clearly indicates an evolutionary relationship between TPMV and members of the genus Morbillivirus. Highest homologies were detected between TPMV and Hendra virus (equine morbillivirus), which recently emerged in Australia, causing outbreaks of fatal respiratory and neurological disease in horses and humans. PMID- 10366581 TI - Interferon action in triply deficient mice reveals the existence of alternative antiviral pathways. AB - Antiviral proteins encoded by the interferon (IFN)-stimulated genes provide a front-line defense against viral infections. In particular, PKR, RNase L, and Mx are considered to be the principal proteins through which IFNs mount an antiviral state. To determine whether alternative antiviral pathways exist, RNase L-/- mice and PKR-/- mice were crossed onto an Mx1(-/-) background to generate a strain of triply deficient (TD) mice. After infections with encephalomyocarditis virus, the TD mice died 3-4 days earlier than infected, wild-type mice. However, there was an IFN dose-dependent increase in survival times after encephalomyocarditis virus infections for both the TD and wild-type mice. Mice that were deficient for PKR or RNase L showed intermediate survival times between those of the TD and wild type mice. Surprisingly, cultured embryonic fibroblasts lacking RNase L, PKR, or both proteins were still able to mount a substantial residual antiviral response against encephalomyocarditis virus or vesicular stomatitis virus after IFN-alpha treatments. These results confirm the antiviral functions of RNase L and PKR in vivo but also provide unequivocal evidence for the existence of novel, innate immune pathways against viruses. PMID- 10366582 TI - Isolation of a human endogenous retroviral HERV-H element with an open env reading frame. AB - About 100 elements of the human endogenous retroviral HERV-H family have full length env genes potentially coding for Env proteins with sequences highly similar to the immunosuppressive peptide CKS-17 from the MLV transmembrane protein p15E. However, previously sequenced HERV-H env genes have contained stop codons or framehifts. To isolate elements with open env reading frames, we first tried to assess the diversity of HERV-H env genes by comparing PCR-generated env sequences from genomic DNA with published HERV-H sequences. A region at the beginning of env displayed a similarity of 84-98% among 15 different elements. We then used a probe from one of the PCR-generated clones, 98% similar to the consensus sequence in this region, to screen a human genomic lambda library. Three HERV-H elements displaying ca. 98% identity in the env gene were isolated and were shown to have integrated relatively recently, after the divergence of the orangutan and the african great ape lineages. One of these elements, HERV H19, had a 1752-bp open env reading frame, producing a 77-kDa Env protein in in vitro translation reactions. This is the first demonstration of a coding competent member of the HERV-H family. These findings raise the possibility that HERV-H Env proteins may play a biological role in human cells. PMID- 10366583 TI - Latent Varicella-zoster virus in human dorsal root ganglia. AB - To understand further the molecular events underlying the process of Varicella zoster virus (VZV) latency in human ganglionic tissues, in situ hybridisation (ISH) for VZV RNA and DNA, and PCR in situ amplification for VZV DNA were used in human dorsal root ganglia from 12 individuals (3 normal and 9 who had died with AIDS). The results showed that (a) two separate regions of the VZV genome, represented by genes 4 and 40, were detected in neurons in two normal and three AIDS ganglia, (b) evidence of transcription of VZV genes 4, 21, 29, and 63 was found in normal and AIDS cases, and (c) VZV DNA and RNA for the same gene (gene 29) was detected in neurons in serial tissue sections in three cases. Thus more than one region of the VZV genome is present in neurons during VZV ganglionic latency, and the presence of both a VZV gene and its corresponding RNA transcript can be shown to occur in the same localised region of DRG tissue. PMID- 10366584 TI - Host cell receptor binding by baculovirus GP64 and kinetics of virion entry. AB - GP64 is the major envelope glycoprotein from budded virions of the baculoviruses Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) and Orgyia pseudotsugata multicapsid nucleopolyhedrovirus (OpMNPV). To examine the potential role of GP64 as a viral attachment protein in host cell receptor binding, we generated, overexpressed, and characterized a soluble form of the OpMNPV GP64 protein, GP64solOp. Assays for trimerization, sensitivity to proteinase K, and reduction by dithiothreitol suggested that GP64solOp was indistinguishable from the ectodomain of the wild-type OpMNPV GP64 protein. Virion binding to host cells was analyzed by incubating virions with cells at 4 degrees C in the presence or absence of competitors, using a single-cell infectivity assay to measure virion binding. Purified soluble GP64 (GP64solOp) competed with a recombinant AcMNPV marker virus for binding to host cells, similar to control competition with psoralen-inactivated wild-type AcMNPV and OpMNPV virions. A nonspecific competitor protein did not similarly inhibit virion binding. Thus specific competition by GP64solOp for virion binding suggests that the GP64 protein is a host cell receptor-binding protein. We also examined the kinetics of virion internalization into endosomes and virion release from endosomes by acid triggered membrane fusion. Using a protease sensitivity assay to measure internalization of bound virions, we found that virions entered Spodoptera frugiperda Sf9 cells between 10 and 20 min after binding, with a half-time of approximately 12.5 min. We used the lysosomotropic reagent ammonium chloride to examine the kinetics of membrane fusion and nucleocapsid release from endosomes after membrane fusion. Ammonium chloride inhibition assays indicated that AcMNPV nucleocapsids were released from endosomes between 15 and 30 min after binding, with a half-time of approximately 25 min. PMID- 10366585 TI - Histidine codons appended to the gene encoding the RPO22 subunit of vaccinia virus RNA polymerase facilitate the isolation and purification of functional enzyme and associated proteins from virus-infected cells. AB - Vaccinia virus encodes a eukaryotic-like RNA polymerase composed of two large and six small subunit protein species. A replication-competent virus with 10 histidine codons added to the single endogenous J4R open reading frame was constructed. The altered migration of the 22-kDa subunit of RNA polymerase on SDS polyacrylamide gel electrophoresis confirmed that J4R encoded the RPO22 subunit and that the mutant virus was genetically stable. The histidine-tagged RNA polymerase bound quantitatively to metal-affinity resins and was eluted in an active form upon addition of imidazole. Glycerol gradient sedimentation of the eluted fraction indicated that most of the RPO22 in infected cells is associated with RNA polymerase. Using stringent washing conditions, metal-affinity chromatography resulted in a several hundred-fold increase in RNA-polymerase specific activity, and substantially pure enzyme was obtained with an additional conventional chromatography step. When mild conditions were used for washing the metal-affinity resin, the vaccinia virus-encoded capping enzyme, early transcription factor, and nucleoside triphosphate phosphohydrolase I specifically co-eluted with the tagged RNA polymerase, consistent with their physical association. The ability to selectively bind RNA polymerase to an affinity column provided a simple and rapid method of concentrating and purifying active enzyme and protein complexes. PMID- 10366586 TI - Differences in the localization and morphology of chromosomes in the human nucleus. AB - Using fluorescence in situ hybridization we show striking differences in nuclear position, chromosome morphology, and interactions with nuclear substructure for human chromosomes 18 and 19. Human chromosome 19 is shown to adopt a more internal position in the nucleus than chromosome 18 and to be more extensively associated with the nuclear matrix. The more peripheral localization of chromosome 18 is established early in the cell cycle and is maintained thereafter. We show that the preferential localization of chromosomes 18 and 19 in the nucleus is reflected in the orientation of translocation chromosomes in the nucleus. Lastly, we show that the inhibition of transcription can have gross, but reversible, effects on chromosome architecture. Our data demonstrate that the distribution of genomic sequences between chromosomes has implications for nuclear structure and we discuss our findings in relation to a model of the human nucleus that is functionally compartmentalized. PMID- 10366587 TI - Intron-independent association of splicing factors with active genes. AB - The cell nucleus is organized as discrete domains, often associated with specific events involved in chromosome organization, replication, and gene expression. We have examined the spatial and functional relationship between the sites of heat shock gene transcription and the speckles enriched in splicing factors in primary human fibroblasts by combining immunofluorescence and fluorescence in situ hybridization (FISH). The hsp90alpha and hsp70 genes are inducibly regulated by exposure to stress from a low basal level to a high rate of transcription; additionally the hsp90alpha gene contains 10 introns whereas the hsp70 gene is intronless. At 37 degrees C, only 30% of hsp90alpha transcription sites are associated with speckles whereas little association is detected with the hsp70 gene, whose constitutive expression is undetectable relative to the hsp90alpha gene. Upon exposure of cells to heat shock, the heavy metal cadmium, or the amino acid analogue azetidine, transcription at the hsp90alpha and hsp70 gene loci is strongly induced, and both hsp transcription sites become associated with speckles in >90% of the cells. These results reveal a clear disconnection between the presence of intervening sequences at specific gene loci and the association with splicing factor-rich regions and suggest that subnuclear structures containing splicing factors are associated with sites of transcription. PMID- 10366588 TI - Transportin-SR, a nuclear import receptor for SR proteins. AB - The SR proteins, a group of abundant arginine/serine (RS)-rich proteins, are essential pre-mRNA splicing factors that are localized in the nucleus. The RS domain of these proteins serves as a nuclear localization signal. We found that RS domain-bearing proteins do not utilize any of the known nuclear import receptors and identified a novel nuclear import receptor specific for SR proteins. The SR protein import receptor, termed transportin-SR (TRN-SR), binds specifically and directly to the RS domains of ASF/SF2 and SC35 as well as several other SR proteins. The nuclear transport regulator RanGTP abolishes this interaction. Recombinant TRN-SR mediates nuclear import of RS domain- bearing proteins in vitro. TRN-SR has amino acid sequence similarity to several members of the importin beta/transportin family. These findings strongly suggest that TRN SR is a nuclear import receptor for the SR protein family. PMID- 10366589 TI - Chs7p, a new protein involved in the control of protein export from the endoplasmic reticulum that is specifically engaged in the regulation of chitin synthesis in Saccharomyces cerevisiae. AB - The Saccharomyces cerevisiae CHS7 gene encodes an integral membrane protein located in the ER which is directly involved in chitin synthesis through the regulation of chitin synthase III (CSIII) activity. In the absence of CHS7 product, Chs3p, but not other secreted proteins, is retained in the ER, leading to a severe defect in CSIII activity and consequently, to a reduced rate of chitin synthesis. In addition, chs7 null mutants show the yeast phenotypes associated with a lack of chitin: reduced mating efficiency and lack of the chitosan ascospore layer, clear indications of Chs7p function throughout the S. cerevisiae biological cycle. CHS3 overexpression does not lead to increased levels of CSIII because the Chs3p excess is retained in the ER. However, joint overexpression of CHS3 and CHS7 increases the export of Chs3p from the ER and this is accompanied by a concomitant increase in CSIII activity, indicating that the amount of Chs7p is a limiting factor for CSIII activity. Accordingly, CHS7 transcription is increased when elevated amounts of chitin synthesis are detected. These results show that Chs7p forms part of a new mechanism specifically involved in Chs3p export from the ER and consequently, in the regulation of CSIII activity. PMID- 10366590 TI - p24 proteins and quality control of LIN-12 and GLP-1 trafficking in Caenorhabditis elegans. AB - Mutations in the Caenorhabditis elegans sel-9 gene elevate the activity of lin-12 and glp-1, which encode members of the LIN-12/NOTCH family of receptors. Sequence analysis indicates SEL-9 is one of several C. elegans p24 proteins. Allele specific genetic interactions suggest that reducing sel-9 activity increases the activity of mutations altering the extracellular domains of LIN-12 or GLP-1. Reducing sel-9 activity restores the trafficking to the plasma membrane of a mutant GLP-1 protein that would otherwise accumulate within the cell. Our results suggest a role for SEL-9 and other p24 proteins in the negative regulation of transport of LIN-12 and GLP-1 to the cell surface, and favor a role for p24 proteins in a quality control mechanism for endoplasmic reticulum-Golgi transport. PMID- 10366591 TI - O-Glycosylation of Axl2/Bud10p by Pmt4p is required for its stability, localization, and function in daughter cells. AB - Cells of the yeast Saccharomyces cerevisiae choose bud sites in a manner that is dependent upon cell type: a and alpha cells select axial sites; a/alpha cells utilize bipolar sites. Mutants specifically defective in axial budding were isolated from an alpha strain using pseudohyphal growth as an assay. We found that a and alpha mutants defective in the previously identified PMT4 gene exhibit unipolar, rather than axial budding: mother cells choose axial bud sites, but daughter cells do not. PMT4 encodes a protein mannosyl transferase (pmt) required for O-linked glycosylation of some secretory and cell surface proteins (Immervoll, T., M. Gentzsch, and W. Tanner. 1995. Yeast. 11:1345-1351). We demonstrate that Axl2/Bud10p, which is required for the axial budding pattern, is an O-linked glycoprotein and is incompletely glycosylated, unstable, and mislocalized in cells lacking PMT4. Overexpression of AXL2 can partially restore proper bud-site selection to pmt4 mutants. These data indicate that Axl2/Bud10p is glycosylated by Pmt4p and that O-linked glycosylation increases Axl2/ Bud10p activity in daughter cells, apparently by enhancing its stability and promoting its localization to the plasma membrane. PMID- 10366592 TI - Isolation, cloning, and localization of rat PV-1, a novel endothelial caveolar protein. AB - By using an immunoisolation procedure (Stan, R.-V., W.G. Roberts, K. Ihida, D. Predescu, L. Saucan, L. Ghitescu, and G.E. Palade. 1997. Mol. Biol. Cell. 8:595 605) developed in our laboratory, we have isolated a caveolar subfraction from rat lung endothelium and we have partially characterized the proteins of this subfraction which include an apparently caveolae-specific glycoprotein we propose to call PV-1 (formerly known as gp68). The isolation and partial sequencing of PV 1, combined with the cloning of the full length PV-1 cDNA led to the following conclusions: (a) PV-1 is a novel single span type II integral membrane protein (438 amino acids long) which forms homodimers in situ; (b) the transmembrane domain of PV-1 is near the NH2 terminus defining a short cytoplasmic endodomain and a large COOH-terminal ectodomain exposed to the blood plasma; (c) PV-1 is N glycosylated and its glycan antennae bear terminal nonreducing galactosyl residues in alpha1-3 linkage. PV-1 is expressed mostly in the lung but both the messenger RNA and the protein can be detected at lower levels also in kidney, spleen, liver, heart, muscle, and brain. No signal could be detected in testis and two lower molecular weight forms were detected in brain. Immunocytochemical studies carried out by immunodiffusion on rat lung with an anti-PV-1 polyclonal antibody directed against a COOH-terminal epitope reveal a specific localization of PV-1 to the stomatal diaphragms of rat lung endothelial caveolae and confirm the extracellular orientation of the PV-1 COOH terminus. PMID- 10366593 TI - A role for ubiquitination in mitochondrial inheritance in Saccharomyces cerevisiae. AB - The smm1 mutation suppresses defects in mitochondrial distribution and morphology caused by the mdm1-252 mutation in the yeast Saccharomyces cerevisiae. Cells harboring only the smm1 mutation themselves display temperature-sensitive growth and aberrant mitochondrial inheritance and morphology at the nonpermissive temperature. smm1 maps to RSP5, a gene encoding an essential ubiquitin-protein ligase. The smm1 defects are suppressed by overexpression of wild-type ubiquitin but not by overexpression of mutant ubiquitin in which lysine-63 is replaced by arginine. Furthermore, overexpression of this mutant ubiquitin perturbs mitochondrial distribution and morphology in wild-type cells. Site-directed mutagenesis revealed that the ubiquitin ligase activity of Rsp5p is essential for its function in mitochondrial inheritance. A second mutation, smm2, which also suppressed mdm1-252 defects, but did not cause aberrant mitochondrial distribution and morphology, mapped to BUL1, encoding a protein interacting with Rsp5p. These results indicate that protein ubiquitination mediated by Rsp5p plays an essential role in mitochondrial inheritance, and reveal a novel function for protein ubiquitination. PMID- 10366594 TI - Morphological differentiation of oligodendrocytes requires activation of Fyn tyrosine kinase. AB - In the central nervous system, myelination of axons occurs when oligodendrocyte progenitors undergo terminal differentiation and initiate process formation and axonal ensheathment. Although it is hypothesized that neuron-oligodendrocyte contact initiates this process, the molecular signals are not known. Here we find that Fyn tyrosine kinase activity is upregulated very early during oligodendrocyte progenitor cell differentiation. Concomitant with this increase is the appearance of several tyrosine phosphorylated proteins present only in differentiated cells. The increased tyrosine kinase activity is specific to Fyn, as other Src family members are not active in oligodendrocytes. To investigate the function of Fyn activation on differentiation, we used Src family tyrosine kinase inhibitors, PP1 and PP2, in cultures of differentiating oligodendrocyte progenitors. Treatment of progenitors with these compounds prevented activation of Fyn and reduced process extension and myelin membrane formation. This inhibition was reversible and not observed with related inactive analogues. A similar effect was observed when a dominant negative Fyn was introduced in progenitor cells. These findings strongly suggest that activation of Fyn is an essential signaling component for the morphological differentiation of oligodendrocytes. PMID- 10366595 TI - Role of phosphorylation sites and the C2 domain in regulation of cytosolic phospholipase A2. AB - Cytosolic phospholipase A2 (cPLA2) mediates agonist-induced arachidonic acid release, the first step in eicosanoid production. cPLA2 is regulated by phosphorylation and by calcium, which binds to a C2 domain and induces its translocation to membrane. The functional roles of phosphorylation sites and the C2 domain of cPLA2 were investigated. In Sf9 insect cells expressing cPLA2, okadaic acid, and the calcium-mobilizing agonists A23187 and CryIC toxin induce arachidonic acid release and translocation of green fluorescent protein (GFP) cPLA2 to the nuclear envelope. cPLA2 is phosphorylated on multiple sites in Sf9 cells; however, only S505 phosphorylation partially contributes to cPLA2 activation. Although okadaic acid does not increase calcium, mutating the calcium binding residues D43 and D93 prevents arachidonic acid release and translocation of cPLA2, demonstrating the requirement for a functional C2 domain. However, the D93N mutant is fully functional with A23187, whereas the D43N mutant is nearly inactive. The C2 domain of cPLA2 linked to GFP translocates to the nuclear envelope with calcium-mobilizing agonists but not with okadaic acid. Consequently, the C2 domain is necessary and sufficient for translocation of cPLA2 to the nuclear envelope when calcium is increased; however, it is required but not sufficient with okadaic acid. PMID- 10366596 TI - A cytoplasmic dynein heavy chain is required for oscillatory nuclear movement of meiotic prophase and efficient meiotic recombination in fission yeast. AB - Meiotic recombination requires pairing of homologous chromosomes, the mechanisms of which remain largely unknown. When pairing occurs during meiotic prophase in fission yeast, the nucleus oscillates between the cell poles driven by astral microtubules. During these oscillations, the telomeres are clustered at the spindle pole body (SPB), located at the leading edge of the moving nucleus and the rest of each chromosome dangles behind. Here, we show that the oscillatory nuclear movement of meiotic prophase is dependent on cytoplasmic dynein. We have cloned the gene encoding a cytoplasmic dynein heavy chain of fission yeast. Most of the cells disrupted for the gene show no gross defect during mitosis and complete meiosis to form four viable spores, but they lack the nuclear movements of meiotic prophase. Thus, the dynein heavy chain is required for these oscillatory movements. Consistent with its essential role in such nuclear movement, dynein heavy chain tagged with green fluorescent protein (GFP) is localized at astral microtubules and the SPB during the movements. In dynein disrupted cells, meiotic recombination is significantly reduced, indicating that the dynein function is also required for efficient meiotic recombination. In accordance with the reduced recombination, which leads to reduced crossing over, chromosome missegregation is increased in the mutant. Moreover, both the formation of a single cluster of centromeres and the colocalization of homologous regions on a pair of homologous chromosomes are significantly inhibited in the mutant. These results strongly suggest that the dynein-driven nuclear movements of meiotic prophase are necessary for efficient pairing of homologous chromosomes in fission yeast, which in turn promotes efficient meiotic recombination. PMID- 10366597 TI - Aip1p interacts with cofilin to disassemble actin filaments. AB - Actin interacting protein 1 (Aip1) is a conserved component of the actin cytoskeleton first identified in a two-hybrid screen against yeast actin. Here, we report that Aip1p also interacts with the ubiquitous actin depolymerizing factor cofilin. A two-hybrid-based approach using cofilin and actin mutants identified residues necessary for the interaction of actin, cofilin, and Aip1p in an apparent ternary complex. Deletion of the AIP1 gene is lethal in combination with cofilin mutants or act1-159, an actin mutation that slows the rate of actin filament disassembly in vivo. Aip1p localizes to cortical actin patches in yeast cells, and this localization is disrupted by specific actin and cofilin mutations. Further, Aip1p is required to restrict cofilin localization to cortical patches. Finally, biochemical analyses show that Aip1p causes net depolymerization of actin filaments only in the presence of cofilin and that cofilin enhances binding of Aip1p to actin filaments. We conclude that Aip1p is a cofilin-associated protein that enhances the filament disassembly activity of cofilin and restricts cofilin localization to cortical actin patches. PMID- 10366598 TI - Induction of filopodia by direct local elevation of intracellular calcium ion concentration. AB - In neuronal growth cones, cycles of filopodial protrusion and retraction are important in growth cone translocation and steering. Alteration in intracellular calcium ion concentration has been shown by several indirect methods to be critically involved in the regulation of filopodial activity. Here, we investigate whether direct elevation of [Ca2+]i, which is restricted in time and space and is isolated from earlier steps in intracellular signaling pathways, can initiate filopodial protrusion. We raised [Ca2+]i level transiently in small areas of nascent axons near growth cones in situ by localized photolysis of caged Ca2+ compounds. After photolysis, [Ca2+]i increased from approximately 60 nM to approximately 1 microM within the illuminated zone, and then returned to resting level in approximately 10-15 s. New filopodia arose in this area within 1-5 min, and persisted for approximately 15 min. Elevation of calcium concentration within a single filopodium induced new branch filopodia. In neurons coinjected with rhodamine-phalloidin, F-actin was observed in dynamic cortical patches along nascent axons; after photolysis, new filopodia often emerged from these patches. These results indicate that local transient [Ca2+]i elevation is sufficient to induce new filopodia from nascent axons or from existing filopodia. PMID- 10366599 TI - A myristoylated calcium-binding protein that preferentially interacts with the Alzheimer's disease presenilin 2 protein. AB - It is well established that mutations in the presenilin 1 and 2 genes cause the majority of early onset Alzheimer's disease (AD). However, our understanding of the cellular functions of the proteins they encode remains rudimentary. Knowledge of proteins with which the presenilins interact should lead to a better understanding of presenilin function in normal and disease states. We report here the identification of a calcium-binding protein, calmyrin, that interacts preferentially with presenilin 2 (PS2). Calmyrin is myristoylated, membrane associated, and colocalizes with PS2 when the two proteins are overexpressed in HeLa cells. Yeast two-hybrid liquid assays, affinity chromatography, and coimmunoprecipitation experiments confirm binding between PS2 and calmyrin. Functionally, calmyrin and PS2 increase cell death when cotransfected into HeLa cells. These results allude to several provocative possibilities for a dynamic role of calmyrin in signaling, cell death, and AD. PMID- 10366600 TI - Monocyte adhesion and spreading on human endothelial cells is dependent on Rho regulated receptor clustering. AB - The GTPase Rho is known to mediate the assembly of integrin-containing focal adhesions and actin stress fibers. Here, we investigate the role of Rho in regulating the distribution of the monocyte-binding receptors E-selectin, ICAM-1, and VCAM-1 in human endothelial cells. Inhibition of Rho activity with C3 transferase or N19RhoA, a dominant negative RhoA mutant, reduced the adhesion of monocytes to activated endothelial cells and inhibited their spreading. Similar effects were observed after pretreatment of endothelial cells with cytochalasin D. In contrast, dominant negative Rac and Cdc42 proteins did not affect monocyte adhesion or spreading. C3 transferase and cytochalasin D did not alter the expression levels of monocyte-binding receptors on endothelial cells, but did inhibit clustering of E-selectin, ICAM-1, and VCAM-1 on the cell surface induced by monocyte adhesion or cross-linking antibodies. Similarly, N19RhoA inhibited receptor clustering. Monocyte adhesion and receptor cross-linking induced stress fiber assembly, and inhibitors of myosin light chain kinase prevented this response but did not affect receptor clustering. Finally, receptor clusters colocalized with ezrin/moesin/ radixin proteins. These results suggest that Rho is required in endothelial cells for the assembly of stable adhesions with monocytes via the clustering of monocyte-binding receptors and their association with the actin cytoskeleton, independent of stress fiber formation. PMID- 10366601 TI - Targeted disruption of the LAMA3 gene in mice reveals abnormalities in survival and late stage differentiation of epithelial cells. AB - Laminin 5 regulates anchorage and motility of epithelial cells through integrins alpha6beta4 and alpha3beta1, respectively. We used targeted disruption of the LAMA3 gene, which encodes the alpha3 subunit of laminin 5 and other isoforms, to examine developmental functions that are regulated by adhesion to the basement membrane (BM). In homozygous null animals, profound epithelial abnormalities were detected that resulted in neonatal lethality, consistent with removal of all alpha3-laminin isoforms from epithelial BMs. Alterations in three different cellular functions were identified. First, using a novel tissue adhesion assay, we found that the mutant BM could not induce stable adhesion by integrin alpha6beta4, consistent with the presence of junctional blisters and abnormal hemidesmosomes. In the absence of laminin 5 function, we were able to detect a new ligand for integrin alpha3beta1 in the epidermal BM, suggesting that basal keratinocytes can utilize integrin alpha3beta1 to interact with an alternative ligand. Second, we identified a survival defect in mutant epithelial cells that could be rescued by exogenous laminin 5, collagen, or an antibody against integrin alpha6beta4, suggesting that signaling through beta1 or beta4 integrins is sufficient for survival. Third, we detected abnormalities in ameloblast differentiation in developing mutant incisors indicating that events downstream of adhesion are affected in mutant animals. These results indicate that laminin 5 has an important role in regulating tissue organization, gene expression, and survival of epithelium. PMID- 10366602 TI - alpha-Dystroglycan is a laminin receptor involved in extracellular matrix assembly on myotubes and muscle cell viability. AB - alpha-Dystroglycan (alpha-DG) is a laminin-binding protein and member of a glycoprotein complex associated with dystrophin that has been implicated in the etiology of several muscular dystrophies. To study the function of DG, C2 myoblasts were transfected stably with an antisense DG expression construct. Myotubes from two resulting clones (11F and 11E) had at least a 40-50% and 80-90% reduction, respectively, in alpha-DG but normal or near normal levels of alpha sarcoglycan, integrin beta1 subunit, acetylcholine receptors (AChRs), and muscle specific kinase (MuSK) when compared with parental C2 cells or three clones (11A, 9B, and 10C) which went through the same transfection and selection procedures but expressed normal levels of alpha-DG. Antisense DG-expressing myoblasts proliferate at the same rate as parental C2 cells and differentiate into myotubes, however, a gradual loss of cells was observed in these cultures. This loss correlates with increased apoptosis as indicated by greater numbers of nuclei with condensed chromatin and more nuclei labeled by the TUNEL method. Moreover, there was no sign of increased membrane permeability to Trypan blue as would be expected with necrosis. Unlike parental C2 myotubes, 11F and 11E myotubes had very little laminin (LN) on their surfaces; LN instead tended to accumulate on the substratum between myotubes. Exogenous LN bound to C2 myotubes and was redistributed into plaques along with alpha-DG on their surfaces but far fewer LN/alpha-DG plaques were seen after LN addition to 11F or 11E myotubes. These results suggest that alpha-DG is a functional LN receptor in situ which is required for deposition of LN on the cell and, further, implicate alpha-DG in the maintenance of myotube viability. PMID- 10366603 TI - A furosemide-sensitive K+-Cl- cotransporter counteracts intracellular Cl- accumulation and depletion in cultured rat midbrain neurons. AB - Efficacy of postsynaptic inhibition through GABAA receptors in the mammalian brain depends on the maintenance of a Cl- gradient for hyperpolarizing Cl- currents. We have taken advantage of the reduced complexity under which Cl- regulation can be investigated in cultured neurons as opposed to neurons in other in vitro preparations of the mammalian brain. Tightseal whole-cell recording of spontaneous GABAA receptor-mediated postsynaptic currents suggested that an outward Cl- transport reduced dendritic [Cl-]i if the somata of cells were loaded with Cl- via the patch pipette. We determined dendritic and somatic reversal potentials of Cl- currents induced by focally applied GABA to calculate [Cl-]i during variation of [K+]o and [Cl-] in the patch pipette. [Cl-]i and [K+]o were tightly coupled by a furosemide-sensitive K+-Cl- cotransport. Thermodynamic considerations excluded the significant contribution of a Na+-K+-Cl- cotransporter to the net Cl- transport. We conclude that under conditions of normal [K+]o the K+-Cl- cotransporter helps to maintain [Cl-]i at low levels, whereas under pathological conditions, under which [K+]o remains elevated because of neuronal hyperactivity, the cotransporter accumulates Cl- in neurons, thereby further enhancing neuronal excitability. PMID- 10366605 TI - Temperature modulation reveals three distinct stages of Wallerian degeneration. AB - After peripheral nerve transection, axons distal to the cut site rapidly degenerate, a process termed Wallerian degeneration. In wild-type mice the compound action potential (CAP) disappears by 3 d. Previous studies have demonstrated that cold temperatures and lower extracellular calcium ion (Ca2+) concentrations can slow the rate of Wallerian degeneration. We have incubated isolated sciatic nerve segments from wild-type and C57BL/Wld mice (which carry a gene slowing Wallerian degeneration) in vitro at 25 and 37 degrees C. At 25 degrees C we found that the degeneration rate of wild-type axons was slowed dramatically, with the CAP preserved up to 7 d post-transection. In contrast, at 37 degrees C the CAPs were minimal at 2 d. When the temperature of wild-type nerves was raised to 37 degrees C after 24-72 hr at 25 degrees C, degeneration occurred within the subsequent 24 hr. Wld nerves, too, were preserved longer at 25 degrees C but, on return to 37 degrees C, degenerated promptly. Cooling the nerve within 12 hr after axotomy enhanced axonal preservation. Neither wild-type nor Wld nerves showed different degeneration rates when they were incubated with 250 microM or 5 or 10 mM extracellular Ca2+ for 1-2 d, suggesting that an abrupt increase in intracellular Ca2+ occurs at the time of axonal destruction. Wallerian degeneration, thus, appears to progress through three distinct stages. Initiation occurs at the time of injury with subsequent temperature-dependent and -independent phases. Nerves appear to remain intact and are able to exclude Ca2+ from entering until an as yet unknown process finally increases axolemmal permeability. PMID- 10366604 TI - Transmembrane domain I contributes to the permeation pathway for serotonin and ions in the serotonin transporter. AB - Mutation of a conserved Asp (D98) in the rat serotonin (5HT) transporter (rSERT) to Glu (D98E) led to decreased 5HT transport capacity, diminished coupling to extracellular Na+ and Cl-, and a selective loss of antagonist potencies (cocaine, imipramine, and citalopram but not paroxetine or mazindol) with no change in 5HT Km value. D98E, which extends the acidic side chain by one carbon, affected the rank-order potency of substrate analogs for inhibition of 5HT transport, selectively increasing the potency of two analogs with shorter alkylamine side chains, gramine, and dihydroxybenzylamine. D98E also increased the efficacy of gramine relative to 5HT for inducing substrate-activated currents in Xenopus laevis oocytes, but these currents were noticeably dependent on extracellular medium acidification. I-V profiles for substrate-independent and -dependent currents indicated that the mutation selectively impacts ion permeation coupled to 5HT occupancy. The ability of the D98E mutant to modulate selective aspects of substrate recognition, to perturb ion dependence as well as modify substrate induced currents, suggests that transmembrane domain I plays a critical role in defining the permeation pathway of biogenic amine transporters. PMID- 10366606 TI - Changes in expression of the DNA repair protein complex DNA-dependent protein kinase after ischemia and reperfusion. AB - Reperfusion of ischemic tissue causes an immediate increase in DNA damage, including base lesions and strand breaks. Damage is reversible in surviving regions indicating that repair mechanisms are operable. DNA strand breaks are repaired by nonhomologous end joining in mammalian cells. This process requires DNA-dependent protein kinase (DNA-PK), composed of heterodimeric Ku antigen and a 460,000 Da catalytic subunit (DNA-PKcs). In this study, a rabbit spinal cord model of reversible ischemia was used to demonstrate the effect of acute CNS injury on the activity and expression of DNA-dependent protein kinase. The DNA binding activity of Ku antigen, analyzed by an electrophoretic mobility shift assay, increased during reperfusion after a short ischemic insult (15 min of occlusion), from which the animals recover neurological function. After severe ischemic injury (60 min of occlusion) and reperfusion that results in permanent paraplegia, Ku DNA binding was reduced. Protein levels of the DNA-PK components Ku70, Ku80, and DNA-PKcs-were monitored by immunoblotting. After 60 min of occlusion, the amount of DNA-PKcs and the enzyme poly(ADP-ribose) polymerase (PARP) decreased with the same time course during reperfusion. Concurrently 150 and 120 kDa fragments were immunostained by an anti-DNA-PKcs monoclonal antibody. This antibody was shown to cross-react with alpha-fodrin breakdown products. The 120 kDa fodrin peptide is associated with caspase-3 activation during apoptosis. Both DNA-PKcs and PARP are also substrates for caspase-3-like activities. The results are consistent with a model in which after a short ischemic insult, DNA repair proteins such as DNA-PK are activated. After severe ischemic injury, DNA damage overwhelms repair capabilities, and cell death programs are initiated. PMID- 10366607 TI - TrkB isoforms with distinct neurotrophin specificities are expressed in predominantly nonoverlapping populations of avian dorsal root ganglion neurons. AB - Alternative splicing of the avian trkB receptor generates an extracellular deletion (ED) isoform missing 11 amino acids from the neurotrophin-binding domain of the full-length (FL) receptor. When expressed in fibroblasts, the ED isoform exhibited restricted neurotrophin specificity compared with that of the FL receptor. Brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and neurotrophin-4 (NT-4) activated the FL receptor, as determined by tyrosine phosphorylation. However, only BDNF was capable of significant activation of the ED isoform, although to a reduced level. Because positively charged residues in NT-3 are important for binding to trkB, two negatively charged aspartate residues within the 11 amino acid motif of FL trkB were mutated to examine the role of electrostatic interactions on ligand binding. As found for the ED isoform, the FL mutated receptor displayed a similar loss of NT-3- and NT-4-mediated activation, in addition to a diminished responsiveness to BDNF. Because of these profound effects on ligand specificity, reverse transcription-PCR was used to understand the expression of the FL and ED receptor isoforms at the level of single neurons. The predominant expression pattern of either FL or ED isoforms in single embryonic DRG neurons establishes the existence of two subpopulations exhibiting differential responsiveness to trkB ligands, indicating that regulated splicing of the extracellular domain of trkB may serve as a mechanism to restrict neuronal responsiveness to the neurotrophins. PMID- 10366608 TI - Characterization of phosphorylation sites on the glutamate receptor 4 subunit of the AMPA receptors. AB - Recent studies have suggested that protein phosphorylation of glutamate receptors may play an important role in synaptic transmission. Specifically, the phosphorylation of AMPA receptors has been implicated in cellular models of synaptic plasticity. The phosphorylation of the glutamate receptor 1 (GluR1) subunit of AMPA receptors by protein kinase A (PKA), protein kinase C (PKC), and Ca2+/calmodulin-dependent protein kinase II (CaMKII) has been characterized extensively. Phosphorylation of this subunit occurs exclusively on the intracellular C-terminal domain. However, the GluR1 subunit C terminus shows low homology to the other AMPA receptor subunits. In this paper we characterized the phosphorylation of AMPA receptor subunit GluR4, using site-specific mutagenesis and biochemical techniques. We found that GluR4 is phosphorylated on serine 842 within the C-terminal domain in vitro and in vivo. Serine 842 is phosphorylated by PKA, PKC, and CaMKII in vitro and is phosphorylated in transfected cells by PKA. Two-dimensional phosphopeptide analysis indicates that serine 842 is the major phosphorylation site on GluR4. In addition, we identified threonine 830 as a potential PKC phosphorylation site. These results suggest that GluR4, which is the most rapidly desensitizing AMPA receptor subunit, may be modulated by phosphorylation. PMID- 10366609 TI - Sequestration of G-protein beta gamma subunits by different G-protein alpha subunits blocks voltage-dependent modulation of Ca2+ channels in rat sympathetic neurons. AB - The membrane-delimited and voltage-dependent inhibition of N-type Ca2+ channels is mediated by Gbeta gamma subunits. Previously, exogenous excess GDP-bound GalphaoA has been shown to dramatically attenuate the norepinephrine (NE) mediated Ca2+ current inhibition by sequestration of Gbeta gamma subunits in rat superior cervical ganglion (SCG) neurons. In the present study, we determined whether the attenuation of NE-mediated modulation is specific to GalphaoA or shared by a number of closely related (Galphatr, GalphaoB, Galphai1, Galphai2, Galphai3, Galphaz) or unrelated (Galphas, Galphaq, Galpha11, Galpha16, Galpha12, Galpha13) Galpha subunits. Individual Galpha subunits from different subfamilies were transiently overexpressed in SCG neurons by intranuclear injection of mammalian expression vectors encoding the desired protein. Strikingly, all Galpha subunits except Galphaz nearly blocked basal facilitation and NE-mediated modulation. Likewise, VIP-mediated Ca2+ current inhibition, which is mediated by cholera toxin-sensitive G-protein, was also completely suppressed by a number of Galpha subunits overexpressed in neurons. Galphas expression produced either enhancement or attenuation of the VIP-mediated modulation-an effect that seemed to depend on the expression level. The onset of the nonhydrolyzable GTP analog, guanylylimidodiphosphate-mediated facilitation was significantly delayed by overexpression of different GDP-bound Galpha subunits. Taken together, these data suggest that a wide variety of Galpha subunits are capable of forming heterotrimers with endogenous Gbeta gamma subunits mediating voltage-dependent Ca2+ channel inhibition. In conclusion, coupling specificity in signal transduction is unlikely to arise as a result of restricted Galpha/Gbeta gamma interaction. PMID- 10366610 TI - Activation and inactivation of the voltage-gated sodium channel: role of segment S5 revealed by a novel hyperkalaemic periodic paralysis mutation. AB - Hyperkalaemic periodic paralysis, paramyotonia congenita, and potassium aggravated myotonia are three autosomal dominant skeletal muscle disorders linked to the SCN4A gene encoding the alpha-subunit of the human voltage-sensitive sodium channel. To date, approximately 20 point mutations causing these disorders have been described. We have identified a new point mutation, in the SCN4A gene, in a family with a hyperkalaemic periodic paralysis phenotype. This mutation predicts an isoleucine-to-phenylalanine substitution at position 1495 located in the transmembrane segment S5 in the fourth homologous domain of the human alpha subunit sodium channel. Introduction of the I1495F mutation into the wild-type channels disrupted the macroscopic current inactivation decay and shifted both steady-state activation and inactivation to the hyperpolarizing direction. The recovery from fast inactivation was slowed, and there was no effect on channel deactivation. Additionally, a significant enhancement of slow inactivation was observed in the I1495F mutation. In contrast, the T704M mutation, a hyperkalaemic periodic paralysis mutation located in the cytoplasmic interface of the S5 segment of the second domain, also shifted activation in the hyperpolarizing direction but had little effect on fast inactivation and dramatically impaired slow inactivation. These results, showing that the I1495F and T704M hyperkalaemic periodic paralysis mutations both have profound effects on channel activation and fast-slow inactivation, suggest that the S5 segment maybe in a location where fast and slow inactivation converge. PMID- 10366611 TI - Regulation of the UNC-18-Caenorhabditis elegans syntaxin complex by UNC-13. AB - The Caenorhabditis elegans unc-13, unc-18, and unc-64 genes are required for normal synaptic transmission. The UNC-18 protein binds to the unc-64 gene product C. elegans syntaxin (Ce syntaxin). However, it is not clear how this protein complex is regulated. We show that UNC-13 transiently interacts with the UNC-18 Ce syntaxin complex, resulting in rapid displacement of UNC-18 from the complex. Genetic and biochemical evidence is presented that UNC-13 contributes to the modulation of the interaction between UNC-18 and Ce syntaxin. PMID- 10366612 TI - Activation of caspase-3 in the retina of transgenic rats with the rhodopsin mutation s334ter during photoreceptor degeneration. AB - The role of caspase-3 in photoreceptor degeneration was examined in a line of transgenic rats that carry a rhodopsin mutation S334ter. Photoreceptor degeneration in these animals is rapid. It is detected as early as postnatal day (PD) 8, and by PD 20, only one of the original 12 rows of nuclei remain in the outer nuclear layer. At PD 11 and 12, the number of photoreceptors dying per day reaches a peak of approximately 30% of the total photoreceptors in the retina. Coincident with this rapid degeneration is an increase in caspase-3-like activity as assessed by the cleavage of a fluorescent substrate N-acetyl-Asp-Glu-Val-Asp aminomethylcoumarin and an increase in activated caspase-3 as determined by Western blot analysis for its 12 kDa subunit. Intraocular injection of an irreversible caspase-3 inhibitor N-benzyloxycarbonal-Asp(OMe)-Glu(OMe)-Val Asp(Ome)-fluoromethyk etone partially protected photoreceptors from degeneration. These findings indicate that a caspase-3-dependent mechanism is operative in photoreceptor death in the transgenic rats under investigation. PMID- 10366613 TI - Activity-dependent modulation of rod photoreceptor cyclic nucleotide-gated channels mediated by phosphorylation of a specific tyrosine residue. AB - Cyclic nucleotide-gated (CNG) channels are crucial for phototransduction in vertebrate rod photoreceptors. The cGMP sensitivity of these channels is modulated by diffusible intracellular messengers, including Ca2+/calmodulin, contributing to negative feedback during sensory adaptation. Membrane-associated protein tyrosine kinases and phosphatases also modulate rod CNG channels, but whether this results from direct changes in the phosphorylation state of the channel protein has been unclear. Here, we show that bovine rod CNG channel alpha subunits (bRET) contain a tyrosine phosphorylation site crucial for modulation. bRET channels expressed in Xenopus oocytes exhibit modulation, whereas rat olfactory CNG channels (rOLF) do not. Chimeric channels reveal that differences in the C terminus, containing the cyclic nucleotide-binding domain, account for this difference. One specific tyrosine in bRET (Y498) appears to be crucial; replacement of this tyrosine in bRET curtails modulation, whereas installation into rOLF confers modulability. As the channel becomes dephosphorylated, there is an increase in the rate of spontaneous openings in the absence of ligand, indicating that changes in the phosphorylation state affect the allosteric gating equilibrium. Moreover, we find that dephosphorylation, which favors channel opening, requires open channels, whereas phosphorylation, which promotes channel closing, requires closed channels. Hence, modulation by changes in tyrosine phosphorylation is activity-dependent and may constitute a positive feedback mechanism, contrasting with negative feedback systems underlying adaptation. PMID- 10366614 TI - Presynaptic mu and delta opioid receptor modulation of GABAA IPSCs in the rat globus pallidus in vitro. AB - The role of enkephalin and the opioid receptors in modulating GABA release within the rat globus pallidus (GP) was investigated using whole-cell patch recordings made from visually identified neurons. Two major GP neuronal subtypes were classified on the basis of intrinsic membrane properties, action potential characteristics, the presence of the anomalous inward rectifier (Ih), and anode break depolarizations. The mu opioid receptor agonist [D-Ala2-N-Me-Phe4-Glycol5] enkephalin (DAMGO) (1 microM) reduced GABAA receptor-mediated IPSCs evoked by stimulation within the striatum. DAMGO also increased paired-pulse facilitation, indicative of presynaptic mu opioid receptor modulation of striatopallidal input. In contrast, the delta opioid agonist D-Pen-[D-Pen2, 5]-enkephalin (DPDPE) (1 microM) was without effect. IPSCs evoked by stimulation within the GP were depressed by application of [methionine 5']-enkephalin (met-enkephalin) (30 microM). Met-enkephalin also reduced the frequency, but not the amplitude, of miniature IPSCs (mIPSCs) and increased paired-pulse facilitation of evoked IPSCs, indicative of a presynaptic action. Both DAMGO and DPDPE reduced evoked IPSCs and the frequency, but not amplitude, of mIPSCs. However, spontaneous action potential-driven IPSCs were reduced in frequency by met-enkephalin and DAMGO, whereas DPDPE was without effect. Overall, these results indicate that presynaptic mu opioid receptors are located on striatopallidal terminals and pallidopallidal terminals of spontaneously firing GP neurons, whereas presynaptic delta opioid receptors are preferentially located on terminals of quiescent GP cells. Enkephalin, acting at both of these receptor subtypes, serves to reduce GABA release in the GP and may therefore act as an adaptive mechanism, maintaining the inhibitory function of the GP in basal ganglia circuitry. PMID- 10366615 TI - Upregulation of surface alpha4beta2 nicotinic receptors is initiated by receptor desensitization after chronic exposure to nicotine. AB - It is hypothesized that desensitization of neuronal nicotinic acetylcholine receptors (nAChRs) induced by chronic exposure to nicotine initiates upregulation of nAChR number. To test this hypothesis directly, oocytes expressing alpha4beta2 receptors were chronically incubated (24-48 hr) in nicotine, and the resulting changes in specific [3H]nicotine binding to surface receptors on intact oocytes were compared with functional receptor desensitization. Four lines of evidence strongly support the hypothesis. (1) The half-maximal nicotine concentration necessary to produce desensitization (9.7 nM) was the same as that needed to induce upregulation (9.9 nM). (2) The concentration of [3H]nicotine for half maximal binding to surface nAChRs on intact oocytes was also similar (11.1 nM), as predicted from cyclical desensitization models. (3) Functional desensitization of alpha3beta4 receptors required 10-fold higher nicotine concentrations, and this was mirrored by a 10-fold shift in concentrations necessary for upregulation. (4) Mutant alpha4beta2 receptors that do not recover fully from desensitization, but not wild-type channels, were upregulated after acute (1 hr) applications of nicotine. Interestingly, the nicotine concentration required for half-maximal binding of alpha4beta2 receptors in total cell membrane homogenates was 20-fold lower than that measured for surface nAChRs in intact oocytes. These data suggest that cell homogenate binding assays may not accurately reflect the in vivo desensitization affinity of surface nAChRs and may account for some of the previously reported differences in the efficacy of nicotine for inducing nAChR desensitization and upregulation. PMID- 10366617 TI - Nerve growth factor signaling through p75 induces apoptosis in Schwann cells via a Bcl-2-independent pathway. AB - Apoptosis is involved in the regulation of Schwann cell numbers during normal development and after axonal damage, but the molecular regulation of Schwann cell death remains unknown. We have used stably transfected rat Schwann cell lines to study the potential roles of nerve growth factor (NGF), the antiapoptotic protein Bcl-2 and the cytokine response modifier A (CrmA) in modulating Schwann cell death in vitro. Bcl-2 inhibited Schwann cell apoptosis induced by survival factor withdrawal, whereas CrmA did not. In contrast, Bcl-2-transfected Schwann cells were susceptible to apoptosis in response to exogenous NGF, whereas CrmA expressing cell lines were resistant. Demonstration of high levels of the low affinity neurotrophin receptor p75 but not the high-affinity TrkA receptor on the Bcl-2-transfected cell lines suggested that the NGF-induced killing was mediated by p75. This was confirmed by resistance of Schwann cells isolated from p75 knockout mice to the NGF-induced cell death. Nerve growth factor also promoted the death of wild-type mouse and rat Schwann cells in the absence of survival factor withdrawal. Endogenous Bcl-2 mRNA was expressed by wild-type Schwann cells in all conditions that promoted survival but was downregulated to undetectable levels after survival factor withdrawal. In conclusion, our results demonstrate the existence of two separate pathways that expedite apoptosis in Schwann cells: a Bcl-2-blockable pathway initiated on loss of trophic support, and a Bcl-2 independent, CrmA-blockable pathway mediated via the p75 receptor. PMID- 10366616 TI - The supporting-cell antigen: a receptor-like protein tyrosine phosphatase expressed in the sensory epithelia of the avian inner ear. AB - After noise- or drug-induced hair-cell loss, the sensory epithelia of the avian inner ear can regenerate new hair cells. Few molecular markers are available for the supporting-cell precursors of the hair cells that regenerate, and little is known about the signaling mechanisms underlying this regenerative response. Hybridoma methodology was used to obtain a monoclonal antibody (mAb) that stains the apical surface of supporting cells in the sensory epithelia of the inner ear. The mAb recognizes the supporting-cell antigen (SCA), a protein that is also found on the apical surfaces of retinal Muller cells, renal tubule cells, and intestinal brush border cells. Expression screening and molecular cloning reveal that the SCA is a novel receptor-like protein tyrosine phosphatase (RPTP), sharing similarity with human density-enhanced phosphatase, an RPTP thought to have a role in the density-dependent arrest of cell growth. In response to hair cell damage induced by noise in vivo or hair-cell loss caused by ototoxic drug treatment in vitro, some supporting cells show a dramatic decrease in SCA expression levels on their apical surface. This decrease occurs before supporting cells are known to first enter S-phase after trauma, indicating that it may be a primary rather than a secondary response to injury. These results indicate that the SCA is a signaling molecule that may influence the potential of nonsensory supporting cells to either proliferate or differentiate into hair cells. PMID- 10366618 TI - Olfactory adaptation depends on the Trp Ca2+ channel in Drosophila. AB - Olfactory adaptation is shown to occur in Drosophila, at both behavioral and physiological levels. In a behavioral paradigm, the extent of adaptation is shown to depend on the dose and duration of the adapting stimulus. Half-maximal adaptation occurred after 15 sec of exposure to an odor, and recovery occurred with a half-time of 1. 5 min, under a set of test conditions. Cross-adaptation was observed among all odor combinations tested, although to a lesser extent than when the same odor was used as both the adapting and the test stimulus. Mutants of the transient receptor potential (Trp) Ca2+ channel were normal in olfactory response, but defective in olfactory adaptation, when measured either behaviorally or in tests of antennal physiology. These results indicate that olfactory response and adaptation can be distinguished. Trp expression was detected in the developing antenna but, surprisingly, not in the mature antenna. These results, together with temperature-shift analysis of a temperature sensitive trp mutant, provide evidence of a role of Trp in olfactory system development. PMID- 10366619 TI - Mapping the agonist binding site of the GABAA receptor: evidence for a beta strand. AB - GABAA receptors, along with the receptors for acetylcholine, glycine, and serotonin, are members of a ligand-gated ion channel superfamily (Ortells and Lunt, 1995). Because of the paucity of crystallographic information for these ligand-gated channels, little is known about the structure of their binding sites or how agonist binding is transduced into channel gating. We used the substituted cysteine accessibility method to obtain secondary structural information about the GABA binding site and to systematically identify residues that line its surface. Each residue from alpha1 Y59 to K70 was mutated to cysteine and expressed with wild-type beta2 subunits in Xenopus oocytes or HEK 293 cells. The sulfhydryl-specific reagent N-biotinylaminoethyl methanethiosulfonate (MTSEA Biotin) was used to covalently modify the cysteine-substituted residues. Receptors with cysteines substituted at positions alpha1 T60, D62, F64, R66, and S68 reacted with MTSEA-Biotin, and alpha1 F64C, R66C, and S68C were protected from reaction by agonist. We conclude that alpha1 F64, R66, and S68 line part of the GABA binding site. The alternating pattern of accessibility of consecutive engineered cysteines to reaction with MTSEA-Biotin indicates that the region from alpha1 Y59 to S68 is a beta-strand. PMID- 10366620 TI - Endocytic active zones: hot spots for endocytosis in vertebrate neuromuscular terminals. AB - We have used a sensitive activity-dependent probe, sulforhodamine 101 (SR101), to view endocytic events within snake motor nerve terminals. After very brief neural stimulation at reduced temperature, SR101 is visualized exclusively at punctate sites located just inside the presynaptic membrane of each terminal bouton. The number of sites (approximately 26 sites/bouton) and their location (in register with postsynaptic folds) are similar to the number and location of active zones in snake motor terminals, suggesting a spatial association between exocytosis and endocytosis under these stimulus conditions. With more prolonged stimulation, larger SR101-containing structures appear at the bouton margins. Thus endocytosis occurs initially at distinct sites, which we call "endocytic active zones," whereas further stimulation recruits a second endocytic paradigm. PMID- 10366621 TI - Expression of human apolipoprotein E3 or E4 in the brains of Apoe-/- mice: isoform-specific effects on neurodegeneration. AB - Apolipoprotein (apo) E isoforms are key determinants of susceptibility to Alzheimer's disease. The apoE4 isoform is the major known genetic risk factor for this disease and is also associated with poor outcome after acute head trauma or stroke. To test the hypothesis that apoE3, but not apoE4, protects against age related and excitotoxin-induced neurodegeneration, we analyzed apoE knockout (Apoe-/-) mice expressing similar levels of human apoE3 or apoE4 in the brain under control of the neuron-specific enolase promoter. Neuronal apoE expression was widespread in the brains of these mice. Kainic acid-challenged wild-type or Apoe-/- mice had a significant loss of synaptophysin-positive presynaptic terminals and microtubule-associated protein 2-positive neuronal dendrites in the neocortex and hippocampus, and a disruption of neurofilament-positive axons in the hippocampus. Expression of apoE3, but not of apoE4, protected against this excitotoxin-induced neuronal damage. ApoE3, but not apoE4, also protected against the age-dependent neurodegeneration seen in Apoe-/- mice. These differences in the effects of apoE isoforms on neuronal integrity may relate to the increased risk of Alzheimer's disease and to the poor outcome after head trauma and stroke associated with apoE4 in humans. PMID- 10366622 TI - A lobster phospholipase C-beta that associates with G-proteins in response to odorants. AB - A cDNA clone encoding a protein of 1116 amino acids with significant homology to beta-isoforms of phospholipase C was isolated from lobster olfactory organ cDNA libraries and named lobPLCbeta. This cDNA hybridized predominantly to a 9 kb transcript in RNA from olfactory organ, pereiopod, brain, and eye-eyestalk and to several smaller minor transcripts only in eye-eyestalk. An antiserum raised to the C terminus of lobPLCbeta detected immunoreactivity in a single 130 kDa band in olfactory aesthetasc hairs, olfactory organ, pereiopod, dactyl, and brain. In eye-eyestalk this 130 kDa band was abundant, and minor bands of 100, 79, and 57 kDa also were detected. In cross sections of the aesthetasc hairs, immunoreactivity was detected in the outer dendritic segments of the olfactory receptor neurons, the site of olfactory transduction. A complex odorant caused lobPLCbeta immunoreactivity to increase in membrane fractions and decrease in soluble fractions of homogenates of aesthetasc hairs. The odorant also increased the amount of lobPLCbeta in immunoprecipitates of Galphaq and Gbeta from homogenates of aesthetasc hairs. These results support the conclusion that lobPLCbeta mediates olfactory transduction. PMID- 10366623 TI - Specification of somatosensory area identity in cortical explants. AB - The H-2Z1 transgene is restricted to a subset of layer IV neurons in the postnatal mouse cortex and delineates exactly the somatosensory area. Expression of the H-2Z1 transgene was used as an areal marker to determine when the parietal cortex becomes committed to a somatosensory identity. We have shown previously that grafts dissected from embryonic day 13.5 (E13.5) H-2Z1 cortex and transplanted into the cortex of nontransgenic newborns express H-2Z1 according to their site of origin. Expression was not modified on heterotopic transplantation (). In the present study, whole cortical explants were isolated at E12.5 from noncortical tissues. The explants developed a regionalized expression of H-2Z1, indicating that regionalization takes place and is maintained in vitro. We used this property and confronted embryonic H-2Z1 cortex with presumptive embryonic sources of regionalizing signals in an in vitro grafting procedure. A great majority of E11.5-E13.5 grafts maintained their presumptive expression of H-2Z1 when grafted heterotopically on nontransgenic E13.5-E15.5 explants. However, a significantly lower proportion of E11.5 parietal grafts expressed H-2Z1 in occipital compared with parietal cortex, indicating that somatosensory identity may be partially plastic at E11.5. Earlier stages could not be tested because the E10.5 grafts failed to develop in vitro. The data suggest that commitment to the expression of a somatosensory area-specific marker coincides with the onset of neurogenesis and occurs well before the birth of the non-GABAergic neurons that express H-2Z1 in vivo. PMID- 10366624 TI - A single growth cone is capable of integrating simultaneously presented and functionally distinct molecular cues during target recognition. AB - A variety of cell recognition pathways affect neuronal target recognition. However, whether such pathways can converge at the level of a single growth cone is not well known. The RP3 motoneuron in Drosophila has previously been shown to respond to the muscle cell surface molecules TOLL and fasciclin III (FAS3), which are normally encountered during RP3 pathfinding in a sequential manner. TOLL and FAS3, putative "negative" and "positive" recognition molecules, respectively, affect RP3 antagonistically. Under normal conditions, TOLL and FAS3 together improve the accuracy of its target recognition. Here, we show that, when presented with concurrent TOLL and FAS3 expression, RP3 responds to both, integrating their effects. This was demonstrated most succinctly by single cell visualization methods. When a balance in relative expression levels between the two antagonistic cues is achieved, the RP3 growth cone exhibits a phenotype virtually identical to that seen when neither TOLL nor FAS3 is misexpressed. Thus, growth cones are capable of quantitatively evaluating distinct recognition cues and integrating them to attain a net result, in effect responding to the "balance of power" between positive and negative influences. We suggest that the ability to integrate multiple recognition pathways in real-time is one important way in which an individual growth cone interprets and navigates complex molecular environments. PMID- 10366625 TI - Abnormalities in neuronal process extension, hippocampal development, and the ventricular system of L1 knockout mice. AB - In humans, mutations in the L1 cell adhesion molecule are associated with a neurological syndrome termed CRASH, which includes corpus callosum agenesis, mental retardation, adducted thumbs, spasticity, and hydrocephalus. A mouse model with a null mutation in the L1 gene (Cohen et al., 1997) was analyzed for brain abnormalities by Nissl and Golgi staining and immunocytochemistry. In the motor, somatosensory, and visual cortex, many pyramidal neurons in layer V exhibited undulating apical dendrites that did not reach layer I. The hippocampus of L1 mutant mice was smaller than normal, with fewer pyramidal and granule cells. The corpus callosum of L1-minus mice was reduced in size because of the failure of many callosal axons to cross the midline. Enlarged ventricles and septal abnormalities were also features of the mutant mouse brain. Immunoperoxidase staining showed that L1 was abundant in developing neurons at embryonic day 18 (E18) in wild-type cerebral cortex, hippocampus, and corpus callosum and then declined to low levels with maturation. In the E18 cortex, L1 colocalized with microtubule-associated protein 2, a marker of dendrites and somata. These new findings suggest new roles for L1 in the mechanism of cortical dendrite differentiation, as well as in guidance of callosal axons and regulation of hippocampal development. The phenotype of the L1 mutant mouse indicates that it is a potentially valuable model for the human CRASH syndrome. PMID- 10366626 TI - GABAA receptor subunit composition and functional properties of Cl- channels with differential sensitivity to zolpidem in embryonic rat hippocampal cells. AB - Using flow cytometry in conjunction with a voltage-sensitive fluorescent indicator dye (oxonol), we have identified and separated embryonic hippocampal cells according to the sensitivity of their functionally expressed GABAA receptors to zolpidem. Immunocytochemical and RT-PCR analysis of sorted zolpidem sensitive (ZS) and zolpidem-insensitive (ZI) subpopulations identified ZS cells as postmitotic, differentiating neurons expressing alpha2, alpha4, alpha5, beta1, beta2, beta3, gamma1, gamma2, and gamma3 GABAA receptor subunits, whereas the ZI cells were neuroepithelial cells or newly postmitotic neurons, expressing predominantly alpha4, alpha5, beta1, and gamma2 subunits. Fluctuation analyses of macroscopic Cl- currents evoked by GABA revealed three kinetic components of GABAA receptor/Cl- channel activity in both subpopulations. We focused our study on ZI cells, which exhibited a limited number of subunits and functional channels, to directly correlate subunit composition with channel properties. Biophysical analyses of GABA-activated Cl- currents in ZI cells revealed two types of receptor-coupled channel properties: one comprising short-lasting openings, high affinity for GABA, and low sensitivity to diazepam, and the other with long-lasting openings, low affinity for GABA, and high sensitivity to diazepam. Both types of channel activity were found in the same cell. Channel kinetics were well modeled by fitting dwell time distributions to biliganded activation and included two open and five closed states. We propose that short- and long-lasting openings correspond to GABAA receptor/Cl- channels containing alpha4beta1gamma2 and alpha5beta1gamma2 subunits, respectively. PMID- 10366627 TI - Netrin-3, a mouse homolog of human NTN2L, is highly expressed in sensory ganglia and shows differential binding to netrin receptors. AB - The netrins comprise a small phylogenetically conserved family of guidance cues important for guiding particular axonal growth cones to their targets. Two netrin genes, netrin-1 and netrin-2, have been described in chicken, but in mouse so far a single netrin gene, an ortholog of chick netrin-1, has been reported. We report the identification of a second mouse netrin gene, which we name netrin-3. Netrin 3 does not appear to be the ortholog of chick netrin-2 but is the ortholog of a recently identified human netrin gene termed NTN2L ("netrin-2-like"), as evidenced by a high degree of sequence conservation and by chromosomal localization. Netrin-3 is expressed in sensory ganglia, mesenchymal cells, and muscles during the time of peripheral nerve development but is largely excluded from the CNS at early stages of its development. The murine netrin-3 protein binds to netrin receptors of the DCC (deleted in colorectal cancer) family [DCC and neogenin] and the UNC5 family (UNC5H1, UNC5H2 and UNC5H3). Unlike chick netrin-1, however, murine netrin-3 binds to DCC with lower affinity than to the other four receptors. Consistent with this finding, although murine netrin-3 can mimic the outgrowth-promoting activity of netrin-1 on commissural axons, it has lower specific activity than netrin-1. Thus, like netrin-1, netrin-3 may also function in axon guidance during development but may function predominantly in the development of the peripheral nervous system and may act primarily through netrin receptors other than DCC. PMID- 10366628 TI - Cell-surface glycoprotein of oligodendrocyte progenitors involved in migration. AB - Myelination by oligodendrocytes in the CNS involves the migration to and recognition and ensheathment of axons. These distinct developmental phases of myelination are assumed to involve the interplay of a precisely regulated set of cell adhesion molecules expressed by both neurons and glial cells. These molecules remain largely unelucidated. In this paper we have identified a large (330 kDa) glycoprotein expressed by murine oligodendrocyte progenitor cells in vitro and in vivo that is downregulated as oligodendrocytes mature. Antigen positive oligodendrocyte progenitor cells purified by panning develop into myelin associated glycoprotein-positive oligodendrocytes and also adhere to cultured neurons. Polyclonal antibodies directed against the protein reduce the migration of oligodendrocyte progenitor cells. The observations suggest that the AN2 antigen may play a role in early stages of myelination. PMID- 10366629 TI - Ephrin-A binding and EphA receptor expression delineate the matrix compartment of the striatum. AB - The striatum integrates limbic and neocortical inputs to regulate sensorimotor and psychomotor behaviors. This function is dependent on the segregation of striatal projection neurons into anatomical and functional components, such as the striosome and matrix compartments. In the present study the association of ephrin-A cell surface ligands and EphA receptor tyrosine kinases (RTKs) with the organization of these compartments was determined in postnatal rats. Ephrin-A1 and ephrin-A4 selectively bind to EphA receptors on neurons restricted to the matrix compartment. Binding is absent from the striosomes, which were identified by mu-opioid receptor immunostaining. In contrast, ephrin-A2, ephrin-A3, and ephrin-A5 exhibit a different mosaic binding pattern that appears to define a subset of matrix neurons. In situ hybridization for EphA RTKs reveals that the two different ligand binding patterns strictly match the mRNA expression patterns of EphA4 and EphA7. Ligand-receptor binding assays indicate that ephrin-A1 and ephrin-A4 selectively bind EphA4 but not EphA7 in the lysates of striatal tissue. Conversely, ephrin-A2, ephrin-A3, and ephrin-A5 bind EphA7 but not EphA4. These observations implicate selective interactions between ephrin-A molecules and EphA RTKs as potential mechanisms for regulating the compartmental organization of the striatum. PMID- 10366630 TI - Impairments in high-frequency transmission, synaptic vesicle docking, and synaptic protein distribution in the hippocampus of BDNF knockout mice. AB - Brain-derived neurotrophic factor (BDNF) promotes long-term potentiation (LTP) at hippocampal CA1 synapses by a presynaptic enhancement of synaptic transmission during high-frequency stimulation (HFS). Here we have investigated the mechanisms of BDNF action using two lines of BDNF knockout mice. Among other presynaptic impairments, the mutant mice exhibited more pronounced synaptic fatigue at CA1 synapses during high-frequency stimulation, compared with wild-type animals. Quantitative analysis of CA1 synapses revealed a significant reduction in the number of vesicles docked at presynaptic active zones in the mutant mice. Synaptosomes prepared from the mutant hippocampus exhibited a marked decrease in the levels of synaptophysin as well as synaptobrevin [vesicle-associated membrane protein (VAMP-2)], a protein known to be involved in vesicle docking and fusion. Treatment of the mutant slices with BDNF reversed the electrophysiological and biochemical deficits in the hippocampal synapses. Taken together, these results suggest a novel role for BDNF in the mobilization and/or docking of synaptic vesicles to presynaptic active zones. PMID- 10366631 TI - Positionally selective growth of embryonic spinal cord neurites on muscle membranes. AB - Motor neurons from distinct positions along the rostrocaudal axis generally innervate muscles or muscle fibers from corresponding axial levels. These topographic maps of connectivity are partially restored after denervation or transplantation under conditions in which factors of timing and proximity are eliminated. It is therefore likely that motor neurons and some intramuscular structures bear cues that bias synapse formation in favor of positionally matched partners. To localize these cues, we studied outgrowth of neurites from embryonic spinal cord explants on carpets of membranes isolated from perinatal rat muscles. Neurites from rostral (cervical) and caudal (lumbar) spinal cord slices exhibit distinct growth preferences. In many instances, rostrally derived neurites grew selectively on membranes from forelimb muscles or from a single thoracic muscle (the serratus anterior) when given a choice between these membranes and membranes from hindlimb muscles or laminin. Caudally derived neurites almost never exhibited such rostral preferences, but instead preferred membranes from hindlimb muscles or a single hindlimb muscle (the gluteus) to rostral muscles or laminin. Likewise, spinal neurites exhibited distinct position-related preferences for outgrowth on membranes of clonal myogenic cell lines derived from specific rostral and caudal muscles. Taken together these results suggest that the membranes of motor axons and myotubes bear complementary labels that vary with rostrocaudal position and regulate neuromuscular connectivity. PMID- 10366632 TI - Spatiotemporal expression patterns of metalloproteinases and their inhibitors in the postnatal developing rat cerebellum. AB - Matrix metalloproteinases (MMPs) are proteolytic enzymes that degrade the components of the extracellular matrix (ECM). The balance between MMPs and their inhibitors [tissue inhibitors of metalloproteinases (TIMPs)] in the pericellular environment determines the most significant proteolytic events in tissue remodeling. In vitro evidence is accumulating that these molecules may be crucial in the maturation of neural cells. Here, we investigated the in vivo expression of MMPs 2, 3, and 9 and TIMPs 1, 2, and 3 in the developing and adult rat cerebellum using immunohistochemistry and in situ hybridization. During postnatal development, all Purkinje (PK) cell somata expressed all the MMPs and TIMPs studied, whereas their growing dendritic trees expressed only MMP 3 and TIMP 3. In the adult, MMP 3 was confined to PK cell bodies, whereas TIMP 3 was expressed in PK cell somata and processes. Irrespective of the developmental stage, Bergmann glial processes contained only MMP 9, but their somata contained both TIMP 1 and MMP 9. In granular cells, MMPs 3 and 9 and TIMPs 1, 2, and 3 were chiefly detected at a time when migration is known to be maximal; except for that of TIMP 1, their expression persisted in the internal granular layer in the adult. The functional relevance of MMP expression was verified by gelatin zymography. MMP 9 activity was maximal on postnatal day 10 (P10) and was detectable at a low level on P15 and in the adult, whereas MMP 2 activity remained similar throughout postnatal development. Regional and cell-specific expression of MMPs and TIMPs closely reflects the successive stages of cerebellar development, thereby suggesting a pivotal role for ECM proteolysis in brain development and plasticity. PMID- 10366633 TI - Propagating activation during oscillations and evoked responses in neocortical slices. AB - Population activity in the cortex is poorly understood. In this report we use voltage-sensitive dye imaging to examine the spatiotemporal patterns of a 7-10 Hz oscillation in neocortical slices from rat somatosensory areas. This oscillation appeared as a component of spontaneous epochs when the preparation was bathed in low [Mg] artificial CSF (ACSF) (Silva et al., 1991). Each epoch started with a synchronized spike, and 3-200 cycles of oscillation emerged afterward. Voltage sensitive dye imaging revealed that the oscillations in the local field potential recordings were actually caused by a propagating population activation. This activation propagated in a relatively uniform size (not expanding). We call this confined, propagating activation a "dynamic ensemble." During each oscillation cycle, one (occasionally two) dynamic ensemble(s) appeared in the slice and was sustained for 60-200 msec. Dynamic ensembles propagated at approximately 30 mm/sec; the activity could propagate in both directions in cortical slices. The propagation consisted in part of "jumps," the locations of which were not fixed. Dynamic ensembles were distinguishable from the epileptiform spikes that occurred in low [Mg] ACSF. Population events similar to dynamic ensembles were also evoked under conditions of unaltered excitability (slice in normal ACSF) by electrical stimulation that activated a low density of neurons in a large area. Our data suggest that self-sustained, spatially confined, and propagating dynamic ensembles might be related to the epoch oscillations in somatosensory cortex seen in vivo (Nicolelis et al., 1995) and thus resemble one form of population activation in the neocortex. PMID- 10366634 TI - Modulation of learning and anxiety by corticotropin-releasing factor (CRF) and stress: differential roles of CRF receptors 1 and 2. AB - The differential modulation of learning and anxiety by corticotropin-releasing factor (CRF) through CRF receptor subtypes 1 (CRFR1) and 2 (CRFR2) is demonstrated. As learning paradigm, context- and tone-dependent fear conditioning of the mouse was used. Injection of CRF into the dorsal hippocampus before training enhanced learning through CRFR1 as demonstrated by the finding that this effect was prevented by the local injection of the unselective CRFR antagonist astressin, but not by the CRFR2-specific antagonist antisauvagine-30 (anti-Svg 30). In contrast, injection of CRF into the lateral intermediate septum impaired learning through CRFR2, as demonstrated by the ability of antisauvagine-30 to block this effect. When antisauvagine-30 was injected alone into the lateral intermediate septum, learning was enhanced. Such tonic control of learning was not observed when astressin or antisauvagine-30 was injected into the dorsal hippocampus. Injection of CRF after the training into the dorsal hippocampus and the lateral intermediate septum also enhanced and impaired learning, respectively. Thus, it was indicated that CRF acted on memory consolidation. It was concluded that the observed effects reflected changes of associative learning and not arousal, attention, or motivation. Although a dose of 20 pmol human/rat CRF was sufficient to affect learning significantly, a fivefold higher dose was required to induce anxiety by injection into the septum. Immobilization for 1 hr generated a stress response that included the induction of anxiety through septal CRFR2 and the subsequent enhancement of learning through hippocampal CRFR1. The involvement of either receptor subtype was demonstrated by region-specific injections of astressin and antisauvagine-30. PMID- 10366635 TI - Attenuation of ischemia-induced cellular and behavioral deficits by X chromosome linked inhibitor of apoptosis protein overexpression in the rat hippocampus. AB - Transient forebrain ischemia produced by four-vessel occlusion (4-VO) triggers the delayed death of CA1 neurons in the hippocampus, resulting in behavioral deficits of spatial learning performance. We demonstrate that CA1 neuronal loss induced by 4-VO (12 min) is preceded by a selective and marked elevation of catalytically active caspase-3 in these neurons, indicative of apoptosis. Virally mediated overexpression of the anti-apoptotic gene X chromosome-linked inhibitor of apoptosis protein (XIAP) prevented both the production of catalytically active caspase-3 and degeneration of CA1 neurons after transient forebrain ischemia. CA1 neurons protected in this manner appeared to function normally, as assessed by immunohistochemical detection of the neuronal activity marker nerve growth factor inducible-A and by spatial learning performance in the Morris water maze. These findings indicate that caspase-3 activation is a key event in ischemic neuronal death and that blockade of this event by XIAP overexpression permits CA1 neurons to survive and operate properly after an ischemic insult. PMID- 10366636 TI - Depression duration but not age predicts hippocampal volume loss in medically healthy women with recurrent major depression. AB - This study takes advantage of continuing advances in the precision of magnetic resonance imaging (MRI) to quantify hippocampal volumes in a series of human subjects with a history of depression compared with controls. We sought to test the hypothesis that both age and duration of past depression would be inversely and independently correlated with hippocampal volume. A sample of 24 women ranging in age from 23 to 86 years with a history of recurrent major depression, but no medical comorbidity, and 24 case-matched controls underwent MRI scanning. Subjects with a history of depression (post-depressed) had smaller hippocampal volumes bilaterally than controls. Post-depressives also had smaller amygdala core nuclei volumes, and these volumes correlated with hippocampal volumes. In addition, post-depressives scored lower in verbal memory, a neuropsychological measure of hippocampal function, suggesting that the volume loss was related to an aspect of cognitive functioning. In contrast, there was no difference in overall brain size or general intellectual performance. Contrary to our initial hypothesis, there was no significant correlation between hippocampal volume and age in either post-depressive or control subjects, whereas there was a significant correlation with total lifetime duration of depression. This suggests that repeated stress during recurrent depressive episodes may result in cumulative hippocampal injury as reflected in volume loss. PMID- 10366637 TI - OCD-Like behaviors caused by a neuropotentiating transgene targeted to cortical and limbic D1+ neurons. AB - To study the behavioral role of neurons containing the D1 dopamine receptor (D1+), we have used a genetic neurostimulatory approach. We generated transgenic mice that express an intracellular form of cholera toxin (CT), a neuropotentiating enzyme that chronically activates stimulatory G-protein (Gs) signal transduction and cAMP synthesis, under the control of the D1 promoter. Because the D1 promoter, like other CNS-expressed promoters, confers transgene expression that is regionally restricted to different D1+ CNS subsets in different transgenic lines, we observed distinct but related psychomotor disorders in different D1CT-expressing founders. In a D1CT line in which transgene expression was restricted to the following D1+ CNS regions-the piriform cortex layer II, layers II-III of somatosensory cortical areas, and the intercalated nucleus of the amygdala-D1CT mice showed normal CNS and D1+ neural architecture but increased cAMP content in whole extracts of the piriform and somatosensory cortex. These mice also exhibited a constellation of compulsive behavioral abnormalities that strongly resembled human cortical-limbic-induced compulsive disorders such as obsessive-compulsive disorder (OCD). These compulsive behaviors included episodes of perseverance or repetition of any and all normal behaviors, repetitive nonaggressive biting of siblings during grooming, and repetitive leaping. These results suggest that chronic potentiation of cortical and limbic D1+ neurons thought to induce glutamatergic output to the striatum causes behaviors reminiscent of those in human cortical-limbic-induced compulsive disorders. PMID- 10366638 TI - Interleukin-1beta immunoreactivity and microglia are enhanced in the rat hippocampus by focal kainate application: functional evidence for enhancement of electrographic seizures. AB - Using immunocytochemistry and ELISA, we investigated the production of interleukin (IL)-1beta in the rat hippocampus after focal application of kainic acid inducing electroencephalographic (EEG) seizures and CA3 neuronal cell loss. Next, we studied whether EEG seizures per se induced IL-1beta and microglia changes in the hippocampus using bicuculline as a nonexcitotoxic convulsant agent. Finally, to address the functional role of this cytokine, we measured the effect of human recombinant (hr)IL-1beta on seizure activity as one marker of the response to kainate. Three and 24 hr after unilateral intrahippocampal application of 0.19 nmol of kainate, IL-1beta immunoreactivity was enhanced in glia in the injected and the contralateral hippocampi. At 24 hr, IL-1beta concentration increased by 16-fold (p < 0.01) in the injected hippocampus. Reactive microglia was enhanced with a pattern similar to IL-1beta immunoreactivity. Intrahippocampal application of 0.77 nmol of bicuculline methiodide, which induces EEG seizures but not cell loss, enhanced IL-1beta immunoreactivity and microglia, although to a less extent and for a shorter time compared with kainate. One nanogram of (hr)IL-1beta intrahippocampally injected 10 min before kainate enhanced by 226% the time spent in seizures (p < 0.01). This effect was blocked by coinjection of 1 microgram (hr)IL-1beta receptor antagonist or 0.1 ng of 3-((+)-2-carboxypiperazin-4-yl)-propyl-1-phosphonate, selective antagonists of IL-1beta and NMDA receptors, respectively. Thus, convulsant and/or excitotoxic stimuli increase the production of IL-1beta in microglia-like cells in the hippocampus. In addition, exogenous application of IL 1beta prolongs kainate-induced hippocampal EEG seizures by enhancing glutamatergic neurotransmission. PMID- 10366639 TI - Differential neural responses during performance of matching and nonmatching to sample tasks at two delay intervals. AB - Visual short-term memory in humans and animals is frequently assessed using delayed matching to sample (DMTS) and delayed nonmatching to sample (DNMTS) tasks across variable delay intervals. Although these tasks depend on certain common mechanisms, there are behavioral differences between them, and neuroimaging provides a means of assessing explicitly whether this is underpinned by differences at a neural level. Findings of delay-dependent deficits, after lesions in humans and animals, suggest that the neural implementation of these tasks may also critically depend on the delay interval. In this study we determined whether there were differential neural responses associated with DMTS and DNMTS tasks at two different delay intervals using functional magnetic resonance imaging. Ten healthy volunteers were studied under four test conditions: DMTS and DNMTS at 5 and 15 sec delay. The main effect of DMTS compared with DNMTS across both delay intervals was associated with significant activation in bilateral head of caudate and medial orbitofrontal cortex. By contrast, DNMTS compared with DMTS was associated with significant activation in mediodorsal thalamus, bilateral lateral orbitofrontal cortex, and left premotor cortex. The main effect of short compared with long delay, across both tasks, was associated with significantly greater activity in occipital and parietal cortices. By contrast, long compared with short delay was associated with significantly greater activity in temporal and ventrolateral frontal cortices. We conclude that DMTS and DNMTS are not equivalent and furthermore that the precise neural implementation of these tasks is a dynamic function of delay interval. PMID- 10366640 TI - Spatial summation in the receptive fields of MT neurons. AB - Receptive fields (RFs) of cells in the middle temporal area (MT or V5) of monkeys will often encompass multiple objects under normal image viewing. We therefore have studied how multiple moving stimuli interact when presented within and near the RF of single MT cells. We used moving Gabor function stimuli, <1 degrees in spatial extent and approximately 100 msec in duration, presented on a grid of possible locations over the RF of the cell. Responses to these stimuli were typically robust, and their small spatial and temporal extent allowed detailed mapping of RFs and of interactions between stimuli. The responses to pairs of such stimuli were compared against the responses to the same stimuli presented singly. The responses were substantially less than the sum of the responses to the component stimuli and were well described by a power-law summation model with divisive inhibition. Such divisive inhibition is a key component of recently proposed "normalization" models of cortical physiology and is presumed to arise from lateral interconnections within a region. One open question is whether the normalization occurs only once in primary visual cortex or multiple times in different cortical areas. We addressed this question by exploring the spatial extent over which one stimulus would divide the response to another and found effective normalization from stimuli quite far removed from the RF center. This supports models under which normalization occurs both in MT and in earlier stages. PMID- 10366641 TI - Single- and multi-whisker channels in the ascending projections from the principal trigeminal nucleus in the rat. AB - This study investigated the relationship between axonal projections and receptive field properties of whisker-sensitive cells in the principal trigeminal sensory nucleus of the rat. The labeling of small groups of trigeminothalamic axons with biotinylated dextran amine disclosed two broad classes of axons; a majority of fibers (68%; n = 107) project to a single barreloid of the ventral posteromedial nucleus, and the remaining group includes axons that innervate both the posterior group of the thalamus and the tectum. Additional terminal sites for axons of this latter group may include the pretectum, the zona incerta, the medial part of the medial geniculate nucleus, and the ventral posteromedial nucleus. Corresponding to these two classes of fibers, 67% of the cells in the principal trigeminal nucleus (n = 313) have single-whisker receptive fields, whereas the rest of the population have receptive fields composed of multiple whiskers. The tonic or phasic properties of the responses apparently bear no relation to the axonal projection patterns. Solid retrograde labeling of cells that project to the ventral posteromedial nucleus and intracellular staining revealed that single whisker cells have small somata and narrow, barrelette-bounded dendritic trees. In contrast, multi-whisker neurons have large multipolar somata, expansive dendritic trees, and many respond antidromically to stimulation of the superior colliculus. Together, these results provide evidence for two main channels of vibrissal information: a single-whisker channel that links trigeminal barrelettes to their corresponding barreloids, and a multi-whisker channel that distributes principally in the posterior group and tectum. PMID- 10366642 TI - Altered serotonin innervation patterns in the forebrain of monkeys treated with (+/-)3,4-methylenedioxymethamphetamine seven years previously: factors influencing abnormal recovery. AB - The recreational drug (+/-)3,4-methylenedioxymethamphetamine (MDMA, "Ecstasy") is a potent and selective brain serotonin (5-HT) neurotoxin in animals and, possibly, in humans. The purpose of the present study was to determine whether brain 5-HT deficits persist in squirrel monkeys beyond the 18-month period studied previously and to identify factors that influence recovery of injured 5 HT axons. Seven years after treatment, abnormal brain 5-HT innervation patterns were still evident in MDMA-treated monkeys, although 5-HT deficits in some regions were less severe than those observed at 18 months. No loss of 5-HT nerve cell bodies in the rostral raphe nuclei was found, indicating that abnormal innervation patterns in MDMA-treated monkeys are not the result of loss of a particular 5-HT nerve cell group. Factors that influence recovery of 5-HT axons after MDMA injury are (1) the distance of the affected axon terminal field from the rostral raphe nuclei, (2) the degree of initial 5-HT axonal injury, and possibly (3) the proximity of damaged 5-HT axons to myelinated fiber tracts. Additional studies are needed to better understand these and other factors that influence the response of primate 5-HT neurons to MDMA injury and to determine whether the present findings generalize to humans who use MDMA for recreational purposes. PMID- 10366643 TI - Gradual emergence of song selectivity in sensorimotor structures of the male zebra finch song system. AB - Birdsong is a model system for understanding how motor and sensory information interact to coordinate behavior. Neurons in one potential site of sensorimotor integration, the forebrain nucleus HVc, have premotor activity during singing and auditory activity during playback of the bird's own song. It is not known whether the high degree of selectivity for learned features of song observed during playback arises in HVc or also in structures afferent to HVc. We recorded in anesthetized adult zebra finches from two structures afferent to HVc: either the nucleus interfacialis (NIf) or the L1 subdivision of the field L complex, and simultaneously from a second electrode in HVc. Correlations in the bursting pattern of ongoing activity of HVc and NIf recordings were observed; these helped to localize the first electrode to NIf recording sites. Most NIf neurons exhibited song-selective responses, but as a population, they were less selective than were HVc neurons. Most L1 neurons were not song-selective. NIf neurons have also been reported to have premotor activity during singing; thus, NIf is another potential site of auditory-motor interactions in the song system. Evidence gathered to date suggests that those brain areas in the passerine forebrain that are recruited during song production also display the most selective learned auditory responses. PMID- 10366644 TI - Basolateral amygdala noradrenergic influences on memory storage are mediated by an interaction between beta- and alpha1-adrenoceptors. AB - Extensive evidence indicates that norepinephrine modulates memory storage through an activation of beta-adrenoceptors in the basolateral nucleus of the amygdala (BLA). Recent findings suggest that the effects of beta-adrenergic activation on memory storage are influenced by alpha1-adrenoceptor stimulation. Pharmacological findings indicate that activation of postsynaptic alpha1-adrenoceptors potentiates beta-adrenoceptor-mediated activation of cAMP formation. The present study examined whether inactivation of alpha1-adrenoceptors in the BLA would alter the dose-response effects on memory storage of intra-BLA infusions of a beta-adrenoceptor agonist, as well as that of a synthetic cAMP analog. Male Sprague Dawley rats received bilateral microinfusions into the BLA of either the beta-adrenoceptor agonist clenbuterol (3-3000 pmol in 0.2 microliter) or 8 bromoadenosine 3':5'-cyclic monophosphate (8-bromo-cAMP) (0.2-7 nmol in 0.2 microliter) alone or together with the alpha1-adrenoceptor antagonist prazosin (0.2 nmol) immediately after training in an inhibitory avoidance task. Retention was tested 48 hr later. Clenbuterol induced a dose-dependent enhancement of retention, and prazosin attenuated the dose-response effects of clenbuterol. Posttraining intra-BLA infusions of 8-bromo-cAMP also induced a dose-dependent enhancement of retention latencies. However, concurrent infusion of prazosin did not alter the dose-response effects of 8-bromo-cAMP. These findings are consistent with the view that alpha1-adrenoceptors affect memory storage by modulating beta-adrenoceptor activation in the BLA. Moreover, these findings are consistent with those of pharmacological studies indicating that beta adrenoceptors modulate memory storage by a direct coupling to adenylate cyclase, whereas alpha1-receptors act indirectly by influencing the beta-adrenoceptor mediated influence on cAMP formation. PMID- 10366645 TI - Activation of NMDA receptors in the suprachiasmatic nucleus produces light-like phase shifts of the circadian clock in vivo. AB - Although there is substantial evidence that glutamate mimics the effects of light on the mammalian circadian clock in vitro, it has been reported that microinjection of glutamate into the suprachiasmatic nucleus of the hypothalamus (SCN) region in vivo does not result in a pattern of phase shifts that mimic those caused by light pulses. The present study was designed to test the hypothesis that microinjection of NMDA into the SCN would induce light-like phase shifts of the circadian clock through activation of the NMDA receptor. Hamsters housed in constant darkness received microinjections of NMDA through guide cannulas aimed at the SCN region at various times throughout the circadian cycle. Wheel running was monitored as a measure of circadian phase. Microinjection of NMDA resulted in circadian phase shifts, the size and direction of which were dependent on the time of injection. The resulting phase-response curve closely resembled that of light. The circadian response showed a clear dose-dependence at circadian time (CT) 13.5 but not at CT19. Both phase delays and advances induced by NMDA were blocked by coinjection of the NMDA antagonist 2-amino-5 phosphopentanoic acid but were slightly attenuated by the non-NMDA antagonist 6 nitro-7-sulfamoylbenzo[f]quinoxaline-2,3-dione disodium. The ability of NMDA to induce phase shifts was not altered by coinjection with tetrodotoxin. These data are consistent with the hypothesis that activation of NMDA receptors is a critical step in the transmission of photic information to the SCN. PMID- 10366647 TI - Brain-derived neurotrophic factor modulates nociceptive sensory inputs and NMDA evoked responses in the rat spinal cord. AB - Central sensitization, the hyperexcitability of spinal processing that often accompanies peripheral injury, is a major component of many persistent pain states. Here we report that the neurotrophin, brain-derived neurotrophic factor (BDNF), is a modulator of excitability within the spinal cord and contributes to the mechanism of central sensitization. BDNF, localized in primary sensory neuron cell bodies and central terminals, potentiates nociceptive spinal reflex responses in an in vitro spinal cord preparation and induces c-fos expression in dorsal horn neurons. NMDA receptor-mediated responses, known as a major contributor to central sensitization, were significantly enhanced by exogenous BDNF. Systemic NGF treatment, a procedure that mimics peripheral inflammatory states, raises BDNF levels in sensory neurons and increases nociceptive spinal reflex excitability. This increased central excitability is reduced by trkB-IgG, a BDNF "antagonist." We also show directly that inflammatory pain-related behavior depends on BDNF release in vivo. Thus behavioral nociceptive responses induced by intraplantar formalin and by intraplantar carageenan are significantly attenuated by trkB-IgG. Hence BDNF is appropriately localized and regulated in inflammatory states and is sufficient and necessary for the expression of central sensitization in the spinal cord. We propose that BDNF may function as a modulator of central sensitization in pathological states, and our results suggest that pharmacological antagonism of BDNF may prove an effective and novel analgesic strategy for the treatment of persistent inflammatory pain states. PMID- 10366648 TI - Association of storage and processing functions in the dorsolateral prefrontal cortex of the nonhuman primate. AB - The prominent role of the prefrontal cortex (PFC) in working memory (WM) is widely acknowledged both in nonhuman primates and in humans. However, less agreement exists on the issue of functional segregation within different subregions of the PFC with regard to the domains of spatial and nonspatial processing or involvement in simpler versus more complex aspects of WM, e.g., maintenance versus processing function. To address these issues, six monkeys were trained to perform four WM tasks that differed with respect to domain (spatial vs nonspatial) and level of WM demand (recall of one vs three items). The delayed response format was used to assess simple one-item memory, whereas self-ordering tasks were used to require the monkey to maintain and organize three items of information within WM. After training, the monkeys received bilateral PFC lesions in one of two different areas, Walker's areas 9 and 8B (dorsomedial convexity; n = 3) or areas 46 and 8A (dorsolateral cortex, n = 3) and then tested postoperatively on all tasks. Monkeys with lesions of the dorsomedial convexity were not impaired either on spatial or nonspatial WM tasks, whether the task required simple storage or sequential processing. By contrast, lesions of the dorsolateral cortex produced a significant and persistent impairment in both simple and complex spatial WM but no impairment in the two nonspatial WM tasks. These results support a functional segregation within the dorsolateral prefrontal cortex for WM: the dorsolateral prefrontal cortex (area 46/8A) is selectively involved in spatial WM, whereas the dorsomedial convexity (area 9/8B) is not critically engaged in either spatial or nonspatial working memory. Furthermore, the specific involvement of area 46/8A in spatial sequencing as well as in single item storage WM tasks supports, in the nonhuman primate, an areal dissociation based on domain rather than on processing demand. PMID- 10366646 TI - Vibrissae-evoked behavior and conditioning before functional ontogeny of the somatosensory vibrissae cortex. AB - The following experiments determined that the somatosensory whisker system is functional and capable of experience-dependent behavioral plasticity in the neonate before functional maturation of the somatosensory whisker cortex. First, unilateral whisker stimulation caused increased behavioral activity in both postnatal day (P) 3-4 and P8 pups, whereas stimulation-evoked cortical activity (14C 2-deoxyglucose autoradiography) was detectable only in P8 pups. Second, neonatal rat pups are capable of forming associations between whisker stimulation and a reinforcer. A classical conditioning paradigm (P3-P4) showed that the learning groups (paired whisker stimulation-shock or paired whisker stimulation warm air stream) exhibited significantly higher behavioral responsiveness to whisker stimulation than controls. Finally, stimulus-evoked somatosensory cortical activity during testing [P8; using 14C 2-deoxyglucose (2-DG) autoradiography] was assessed after somatosensory conditioning from P1-P8. No learning-associated differences in stimulus-evoked cortical activity were detected between learning and nonlearning control groups. Together, these experiments demonstrate that the whisker system is functional in neonates and capable of experience-dependent behavioral plasticity. Furthermore, in contrast to adult somatosensory classical conditioning, these data suggest that the cortex is not required for associative somatosensory learning in neonates. PMID- 10366649 TI - Differential regulation of mPER1 and mTIM proteins in the mouse suprachiasmatic nuclei: new insights into a core clock mechanism. AB - Recent discoveries have identified a framework for the core circadian clock mechanism in mammals. Development of this framework has been based entirely on the expression patterns of so-called "clock genes" in the suprachiasmatic nuclei (SCN), the principal clock of mammals. We now provide data concerning the protein expression patterns of two of these genes, mPer1 and mTim. Our studies show that mPER1 and mTIM are nuclear antigens expressed in the SCN and extensively throughout the forebrain. Expression of mPER1 in the SCN was rhythmic under entrained conditions and with clear circadian cycling under free-running conditions. Expression of mPER1 elsewhere in the mouse forebrain was not rhythmic. In contrast to mPER1, mTIM expression in the SCN did not vary with time in mice housed in either a light/dark cycle or in constant dim red light. The phase relationship between mPer1 RNA and mPER1 cycles in the SCN is consistent with a negative feedback model of the mammalian clock. The invariant nature of nuclear mTIM in the SCN suggests that its participation in negative feedback occurs only after mPER1 has entered the nucleus, and that the abundance of mTIM is not regulated by the circadian clock or the light/dark cycle. PMID- 10366650 TI - GATA-3 is involved in the development of serotonergic neurons in the caudal raphe nuclei. AB - The GATA-3 transcription factor shows a specific and restricted expression pattern in the developing and adult mouse brain. In the present study we investigated the role of GATA-3 in the caudal raphe system, which is known to operate as a modulator of motor activity. We demonstrate that virtually all neurons in the caudal raphe nuclei that express GATA-3 also produce serotonin. Absence of GATA-3, as analyzed in chimeric -/- mice, affects the cytoarchitecture of serotonergic neurons in the caudal raphe nuclei. As a result the chimeras show a serious defect in their locomotor performance on a rotating rod. In sum, we conclude that GATA-3 plays a major role in the development of the serotonergic neurons of the caudal raphe nuclei, and that it is crucial for their role in locomotion. PMID- 10366651 TI - Ultrastructural correlates of quantal synaptic function at single CNS synapses. AB - We have tested the hypothesis that functional differences between synapses are associated with ultrastructure in cultured cortical neurons. Using Ca(2+) imaging, we measured NMDA receptor-mediated miniature synaptic calcium transients attributed to the spontaneous release of single transmitter quanta. After imaging, the identified neurons were processed for serial transmission electron microscopy. At sites of quantal NMDA receptor-dependent Ca(2+) transients, we confirmed the presence of excitatory synapses and measured spine size and synaptic contact area. Our results demonstrate that synapse size correlates positively with the amplitude of the NMDA receptor-mediated postsynaptic response, suggesting that larger synapses express a greater number of NMDA receptors. Therefore, regulation of quantal amplitude may involve processes that alter synapse size. PMID- 10366652 TI - Direct alteration of the P/Q-type Ca2+ channel property by polyglutamine expansion in spinocerebellar ataxia 6. AB - Spinocerebellar ataxia 6 (SCA6) is caused by expansion of a polyglutamine stretch, encoded by a CAG trinucleotide repeat, in the human P/Q-type Ca(2+) channel alpha(1A) subunit. Although SCA6 shares common features with other neurodegenerative glutamine repeat disorders, the polyglutamine repeats in SCA6 are exceptionally small, ranging from 21 to 33. Because this size is too small to form insoluble aggregates that have been blamed for the cause of neurodegeneration, SCA6 is the disorder suitable for exploring the pathogenic mechanisms other than aggregate formation, whose universal role has been questioned. To characterize the pathogenic process of SCA6, we studied the effects of polyglutamine expansion on channel properties by analyzing currents flowing through the P/Q-type Ca(2+) channels with an expanded stretch of 24, 30, or 40 polyglutamines, recombinantly expressed in baby hamster kidney cells. Whereas the Ca(2+) channels with ca. 54.4 MPa): Kp,lc approximately 3x10(3) vs. Kp,gel approximately 6x10(3). The presence of sterols (45% molar ergosterol or cholesterol) caused an increase in the partition coefficients, regardless of pressure, ergosterol having a more pronounced effect (Kp approximately 2x10(4) 6x10(4)). Kp was pressure dependent in both cases, but mainly with cholesterol (Kp approximately 2x10(3)-2x10(4)). At variance with cholesterol, when ergosterol was used, no phase transition was detected. This difference cannot be due to a more extended uptake of filipin by cholesterol-containing membranes, and so must be due to specific interactions with cholesterol. In agreement with this finding, we discovered that filipin is more tightly packed (lower partial molar volume) in the cholesterol-rich phase than in the ergosterol-rich phase. Our results also point to a 2:1 DPPC:cholesterol stoichiometry in the cholesterol-rich phase (17% molar cholesterol). All partition coefficients were calculated from steady-state fluorescence anisotropy measurements. PMID- 10366666 TI - Nucleoside transport in human colonic epithelial cell lines: evidence for two Na+ independent transport systems in T84 and Caco-2 cells. AB - RT-PCR of RNA isolated from monolayers of the human colonic epithelial cell lines T84 and Caco-2 demonstrated the presence of mRNA for the two cloned Na+ independent equilibrative nucleoside transporters, ENT1 and ENT2, but not for the cloned Na+-dependent concentrative nucleoside transporters, CNT1 and CNT2. Uptake of [3H]uridine by cell monolayers in balanced Na+-containing and Na+-free media confirmed the presence of only Na+-independent nucleoside transport mechanisms. This uptake was decreased by 70-75% in the presence of 1 microM nitrobenzylthioinosine, a concentration that completely inhibits ENT1, and was completely blocked by the addition of 10 microM dipyridamole, a concentration that inhibits both ENT1 and ENT2. These findings indicate the presence in T84 and Caco-2 cells of two functional Na+-independent equilibrative nucleoside transporters, ENT1 and ENT2. PMID- 10366667 TI - Purification and characterization of a major zinc binding protein from renal brush border membrane of rat. AB - In spite of the fact that zinc is an essential trace element, mechanisms that contribute to zinc homeostasis in mammals are poorly understood. An attempt has been made to identify and purify zinc binding components from renal brush border membrane (BBM), which could be involved in the binding of zinc and the subsequent translocation across the BBM. A 40 kDa major zinc binding protein has been identified and purified from renal BBM, which showed a dissociation constant (Kd) of 211 microM and maximal binding (Bmax) of 207 nmol/mg protein. 8 g zinc atoms could interact with 1 mol of protein. Specificity of the protein for zinc was checked by metal displacement and UV-absorption assay. It was found that only Cd2+ could displace the zinc bound to the protein. Other metals tested (Cu2+, Mg2+, Ca2+) did not show any interaction with the protein. These data indicated that purified protein is highly specific and has a high affinity for zinc. The carbohydrate content was found to be 7.85 mg% in the purified protein. Immunofluorescence localization of this protein in kidney sections revealed that this major zinc binding protein is exclusively localized in the proximal convoluted tubules. These results suggested that the 40 kDa major zinc binding transmembrane glycoprotein is highly specific for zinc and has a high affinity for zinc. PMID- 10366668 TI - Extracellular ATP-induced inward current in isolated epithelial cells of the endolymphatic sac. AB - Using whole-cell patch-clamp technique and Fura-2 fluorescence measurement, the presence of ATP-activated ion channels and its dependence on intracellular Ca2+ concentration ([Ca2+]i) in the epithelial cells of the endolymphatic sac were investigated. In zero current-clamp configuration, the average resting membrane potential was -66.8+/-1.3 mV (n=18). Application of 30 microM ATP to the bath induced a rapid membrane depolarization by 43.1+/-2.4 mV (n=18). In voltage-clamp configuration, ATP-induced inward current at holding potential (VH) of -60 mV was 169.7+/-6.3 pA (n=18). The amplitude of ATP-induced currents increased in sigmoidal fashion over the concentration range between 0.3 and 300 microM with a Hill coefficient (n) of 1.2 and a dissociation constant (Kd) of 11.7 microM. The potency order of purinergic analogues in ATP-induced current, which was 2MeSATP>ATPgammas>/=ATP>alpha, beta-ATP>ADP=AMP>/=adenosine=UTP, was consistent with the properties of the P2Y receptor. The independence of the reversal potential of the ATP-induced current from Cl- concentration suggests that the current is carried by a cation channel. The relative ionic permeability ratio of the channel modulated by ATP for cations was Ca2+>Na+>Li+>Ba2+>Cs+=K+. ATP (10 microM) increased [Ca2+]i in an external Ca2+-free solution to a lesser degree than that in the external solution containing 1.13 mM CaCl2. ATP-induced increase in [Ca2+]i can be mimicked by application of ionomycin in a Ca2+-free solution. These results indicate that ATP increases [Ca2+]i through the P2Y receptor with a subsequent activation of the non-selective cation channel, and that these effects of ATP are dependent on [Ca2+]i and extracellular Ca2+. PMID- 10366669 TI - Interaction of the anti-cancer drug cisplatin with phosphatidylserine in intact and semi-intact cells. AB - The anti-cancer drug cisplatin (cis-diamminedichloroplatinum(II)) forms a stable coordination complex with phosphatidylserine (PS) in model membrane systems (Speelmans et al., Biochemistry 36 (1997) 10545-10550). Because a similar interaction in vivo would be expected to have important physiological implications we studied cisplatin-PS interaction in human erythrocytes and tumor cell lines. Although cisplatin was efficiently taken up by intact erythrocytes, a cisplatin-PS complex was only detected in cells which had lysed as a result of prolonged storage or hypotonic shock. Despite the use of highly sensitive detection methods, and despite efficient cellular uptake of cisplatin, a complex could also not be detected in four human tumor cell lines, unless cells were permeabilized. In experiments in which cisplatin was incubated with PS-containing liposomes in the presence of an alternative cellular substrate, such as reduced glutathione, the relative affinity of cisplatin for PS was found to be low. Moreover, loading erythrocyte ghosts with physiological concentrations of glutathione strongly reduced cisplatin-PS complexation. Thus, in intact (tumor) cells a complex is not detected, most likely, because of the presence of higher affinity substrates. Though a transient complexation of cisplatin to PS cannot be excluded, our data suggest that cisplatin-PS does not play a direct role in the cellular (cyto)toxicity of cisplatin. PMID- 10366670 TI - Calcium dependence of Pi phosphorylation of sarcoplasmic reticulum Ca2+-ATPase at low water content: water dependence of the E2-->E1 conversion. AB - Enzymes entrapped in reverse micelles can be studied in low-water environments that have the potential of restricting conformational mobility in specific steps of the reaction cycle. Sarcoplasmic reticulum Ca2+-ATPase was incorporated into a reverse-micelle system (TPT) composed of toluene, phospholipids, Triton X-100 and varying amounts of water (0.5-7%, v/v). Phosphorylation of the Ca2+-ATPase by ATP required the presence of both water and Ca2+ in the micelles. No phosphoenzyme (EP) was detected in the presence of EGTA. Phosphorylation by Pi (inorganic phosphate) in the absence of Ca2+ was observed at water content below that necessary for phosphorylation by ATP. In contrast to what is observed in a totally aqueous medium, EP formed by Pi was partially resistant to dephosphorylation by Ca2+. However, the addition of non-radioactive Pi to the EP already formed caused a rapid decrease in radiolabelled enzymes, as expected for the isotopic dilution, indicating the existence of an equilibrium (E+Pi<-->EP). Phosphorylation by Pi also occurred in TPT containing millimolar Ca2+ concentrations in a range of water concentrations (2-5% v/v). The substrates p nitrophenyl phosphate, acetyl phosphate, ATP and GTP increased the EP level under these conditions. These results suggest that: (1) the rate of conversion of the ATPase conformer E2 into E1 is greatly reduced at low water content, so that E2- >E1 becomes the rate-limiting step of the catalytic cycle; and (2) in media of low water content, Pi can phosphorylate both E1Ca and E2. Thus, the effect of enzyme hydration is complex and involves changes in the phosphorylation reaction at the catalytic site, in the equilibrium between E2 and E1 conformers, and in their specificity for substrates. PMID- 10366671 TI - Rapid diffusion of spectrin bound to a lipid surface. AB - Human erythrocyte spectrin was labelled with the probe 5, 5'-disulfato-1-(6 hexanoic acid N-hydroxysuccinimide ester)-1'-ethyl-3,3,3',3' tetramethylindocarbocyanine (Cy3). Cy3-spectrin was bound to the outer surface of dimyristoylphosphatidylcholine (DMPC) multilamellar vesicles and its diffusion measured by fluorescence recovery after photobleaching (FRAP). It was found that at 30 degrees C, above the lipid gel to liquid-crystalline phase transition of the lipids, Cy3-spectrin had an unexpectedly high diffusion coefficient D=(2.1+/ 0.6)x10(-7)) cm2/s. At the phase transition, diffusion of Cy3-spectrin was only slightly lower; D=(1.3+/-0.3)x10(-7) cm2/s, whereas at 14 degrees C, well below the lipid phase transition, diffusion was found to be much slower with D=(3.1+/ 0.12)x10(-9) cm2/s. The fast diffusion of Cy3-spectrin on the lipid surface implies that the individual bonds which bind spectrin to the lipid surface must rapidly be made and broken. In the light of these results, spectrin-lipid interactions alone appear unlikely to have any significant role in supporting the cell membrane. Probably, the interactions serve only to localise the spectrin at the inner lipid surface in order to facilitate formation of the cytoskeleton. PMID- 10366672 TI - Substitution of the N-terminal segment of the plasma membrane Ca pump isoform 4 by that of isoform 1 results in a fully functional chimeric enzyme. AB - The N-terminal segment of the plasma membrane Ca2+ pump (PMCA) is one of the most variable regions among the four isoforms of the enzyme and its functional importance is unknown. In the present work, the N-terminal segment of the highly active C-terminally truncated h4 mutant, h4(ct120) was modified either by substituting residues 18-43 by residues 43-75 or by replacing residues 1-75 by the homologous region from isoform h1 (residues 1-79). Immunoblot analysis of microsomal membranes from transfected COS-1 cells showed that the two N terminally mutated proteins were correctly expressed at a level similar to that of h4(ct120). Measurements of the Ca2+ uptake by microsomal vesicles from transfected COS-1 cells indicated that mutant (18-43-->43-75)h4(ct120) had only negligible Ca2+ transport activity while the chimeric (n1-79)h1h4(ct120) enzyme was fully capable of functioning as a calcium pump. Like h4(ct120), the chimeric mutant was not stimulated further by calmodulin, and was inhibited to a similar degree by the C28R2 peptide corresponding to the calmodulin binding autoinhibitory region of the pump. Moreover, the apparent affinity for Ca2+ and the ATP dependence of the chimeric enzyme were similar to those of the h4(ct120) pump suggesting that the variability of sequence between the N-terminal segment of PMCA isoforms h1 and h4 involves amino acid substitutions that do not substantially change the behavior of the h4 enzyme. Altogether, these results demonstrate that for activity the h4 Ca pump requires a specific amino acid sequence at its N-terminus, and the essential elements for a fully active enzyme can be provided by the N-terminal segment of isoform h1 despite the variability. PMID- 10366673 TI - Interactions of sodium pentobarbital with D-glucose and L-sorbose transport in human red cells. AB - Pentobarbital acts as a mixed inhibitor of net D-glucose exit, as monitored photometrically from human red cells. At 30 degrees C the Ki of pentobarbital for inhibition of Vmax of zero-trans net glucose exit is 2.16+/-0.14 mM; the affinity of the external site of the transporter for D-glucose is also reduced to 50% of control by 1. 66+/-0.06 mM pentobarbital. Pentobarbital reduces the temperature coefficient of D-glucose binding to the external site. Pentobarbital (4 mM) reduces the enthalpy of D-glucose interaction from 49.3+/-9.6 to 16.24+/-5.50 kJ/mol (P<0.05). Pentobarbital (8 mM) increases the activation energy of glucose exit from control 54.7+/-2.5 kJ/mol to 114+/-13 kJ/mol (P<0.01). Pentobarbital reduces the rate of L-sorbose exit from human red cells, in the temperature range 45 degrees C-30 degrees C (P<0.001). On cooling from 45 degrees C to 30 degrees C, in the presence of pentobarbital (4 mM), the Ki (sorbose, glucose) decreases from 30.6+/-7.8 mM to 14+/-1.9 mM; whereas in control cells, Ki (sorbose, glucose) increases from 6.8+/-1.3 mM at 45 degrees C to 23.4+/-4.5 mM at 30 degrees C (P<0.002). Thus, the glucose inhibition of sorbose exit is changed from an endothermic process (enthalpy change=+60.6+/-14.7 kJ/mol) to an exothermic process (enthalpy change=-43+/-6.2 7 kJ/mol) by pentobarbital (4 mM) (P<0.005). These findings indicate that pentobarbital acts by preventing glucose-induced conformational changes in glucose transporters by binding to 'non-catalytic' sites in the transporter. PMID- 10366674 TI - Hypertonicity stimulates taurine uptake and transporter gene expression in Caco-2 cells. AB - The osmoregulation of taurine transport in intestinal epithelial cells was investigated using human intestinal Caco-2 cells. The activity of taurine transport in the Caco-2 cells was increased by hypertonic conditions. This hypertonicity-induced up-regulation was dependent on both the culturing time and the osmotic pressure. Hypertonicity did not affect the activity of L-leucine, L lysine, or L-glutamic acid transport, suggesting that osmoregulation was specific to taurine transport. The intracellular taurine content of Caco-2 cells was also increased by culturing in a hypertonic medium. These hypertonicity-induced changes in the intracellular taurine content and transport activity were reversible. A kinetic analysis of taurine transport in the control and hypertonic cells suggested that the up-regulation was associated with an increase in the amount of the taurine transporter. The mRNA level of the taurine transporter in hypertonic cells was markedly higher than that in the control cells, indicating that this osmotic regulation was due to the increased expression of the taurine transporter gene. PMID- 10366675 TI - Homeostasis of the membrane proton permeability in Bacillus subtilis grown at different temperatures. AB - Bacillus subtilis was grown at its growth temperature limits and at various temperatures in between the lower and upper growth temperature boundary. Liposomes were made of the extracted membrane lipids derived from these cells. The headgroup composition of the cytoplasmic membrane lipids did not differ significantly at the lower (13 degrees C) and upper (50 degrees C) temperature boundary. The averaged lipid acyl chain length, degree of saturation, and ratio of iso- and anteiso-branched fatty acids increased with the temperature. At the temperature of growth, the membranes were in a liquid-crystalline phase, but liposomes derived from cells grown at 13 degrees C were almost threefold more viscous than those derived from 50 degrees C grown cells. The temperature dependence of the proton permeability of the liposomes was determined using the acid-pulse method with monitoring of the outside pH with the fluorescent probe pyranine. The proton permeability of each liposome preparation increased with the temperature. However, the proton permeability of the liposomes at the growth temperature of the cells from which the lipids were derived was almost constant. These data indicate that the growth temperature dependent variation in lipid acyl chain composition permits maintenance of the proton permeability of the cytoplasmic membrane. This 'homeo-proton permeability adaptation' precludes futile cycling of protons at higher growth temperatures and allows cells to sustain the proton motive force as a driving force for essential energy transducing processes. PMID- 10366676 TI - Subunit D of the vacuolar H+-ATPase of Arabidopsis thaliana. AB - A 1034 bp cDNA encoding the full length sequence of subunit D of the vacuolar H+ ATPase was cloned from Arabidopsis thaliana. The open reading frame of the cDNA clone vatpD contains 780 bp and codes for a protein of 29.1 kDa with a pI of 9.52. Structural predictions show similarities to subunit gamma of the F-ATP synthases. Identity between subunit D of the vacuolar H+-ATPase of A. thaliana and subunits D from other eukaryotic organisms is in the range of 57% (Bos taurus) to 48% (Candida albicans). Hybridization of genomic DNA with vatpD indicates the existence of one gene copy of subunit D in A. thaliana. Northern blot hybridization and in situ hybridization showed expression of vatpD in all cell types. The expression of subunit D was not modified by salt stress or abscisic acid treatment in A. thaliana. PMID- 10366677 TI - Sensitization of norepinephrine release in medial prefrontal cortex: effect of different chronic stress protocols. AB - Previously, we demonstrated that continuous exposure of rats to cold (5 degrees C) for 2-3 weeks potentiates the increase in extracellular norepinephrine in the medial prefrontal cortex produced by acute tail shock. In the present study, we used in vivo microdialysis to examine whether this sensitization of evoked norepinephrine release also occurs in the medial prefrontal cortex following exposure to other chronic stress protocols. Rats exposed to 30 min of intermittent foot shock (0.6 mA) each day for 14 days, did not exhibit a greater increase in extracellular norepinephrine in response to acute tail shock. To determine whether this discrepancy between cold exposure and foot shock might be related to differences in the nature or the pattern of exposure to the chronic stressor, we also examined the effect of intermittent exposure to cold or continuous exposure to a foot shock protocol on tail shock-evoked norepinephrine release. Sensitized norepinephrine release did not develop following either intermittent exposure to cold (5 degrees C; 4 h/day for 14 days) or continuous exposure to a foot shock protocol (0.6 mA trains at random intervals 24 h/day for 14 days), suggesting that both the nature of the stressor as well as the pattern of exposure to the chronic stressor play a role in the development of sensitized norepinephrine release. PMID- 10366678 TI - Characterization of the neuroprotective and toxic effects of alpha7 nicotinic receptor activation in PC12 cells. AB - The alpha7 nicotinic receptor partial agonist DMXB protected differentiated PC12 cells from NGF+ serum deprivation over a concentration range (1-10 microM) that correlated with activation of protein kinase C. Increased toxicity was observed at a higher concentration of DMXB (30 microM) that did not elevate protein kinase C activity, but did increase tyrosine protein kinase activity. Neuroprotection was blocked with the protein kinase C-inhibitor bis-indolemaleimide, while toxicity was attenuated with the tyrosine protein kinase-antagonists herbimycin and genistein. The alpha7-selective antagonist methyllyconitine attenuated both the protective and toxic actions of DMXB, but in temporally distinct manners. Methyllyconitine (1 microM) attenuated toxicity when added 10 s before, but not 10 s after, 30 microM DMXB. In contrast, it blocked neuroprotection when added 10 min post-agonist addition. This temporal difference in receptor-activation that was necessary for protection vs. toxicity reflected the time courses for agonist induced desensitization of the receptor expressed in Xenopus oocytes. These results indicate that alpha7 nicotinic receptors act through different intracellular transduction processes to protect or kill cells. Further, they suggest that the transduction processes may be differentially activated depending on the amplitude and duration of calcium signals. PMID- 10366679 TI - Distribution of cyclooxygenase-1 and cyclooxygenase-2 mRNAs and proteins in human brain and peripheral organs. AB - We used the techniques of reverse transcriptase-polymerase chain reaction, Western blotting and immunohistochemistry to evaluate the expression of cyclooxygenase (COX)-1 and -2 in brain and peripheral organs of Alzheimer disease (AD) and control cases. We found both COX-1 and COX-2 to be constitutively expressed in all organs tested, i.e., brain, heart, liver, kidney, spleen and intestine. COX-2 was substantially upregulated in affected areas of AD brain and in infarcted areas of human heart. COX-1 was only mildly upregulated in AD brain. Immunohistochemically, COX-2 was strongly expressed in the perinuclear, dendritic and axonal areas of pyramidal neurons, with enhanced staining in AD. These data suggest a special role for COX-2 in neuronal function. PMID- 10366680 TI - Tyrosine hydroxylase, aromatic L-amino acid decarboxylase and dopamine metabolism after chronic treatment with dopaminergic drugs. AB - Mice were treated with dopamine (DA) receptor agonist and antagonist drugs: Agonists: (+/-)-SKF 38393 ((+/-)-1-phenyl-2,3,4, 5-tetrahydro-(1H)-3-benzazepine 7,8-diol) [DA D1-like]; bromocriptine, [DA D2 selective]; quinpirole, [DA D2/D3 preferring]; (+/-)-7-hydroxy-dipropylamino-tetralin (7-OH-DPAT), [DA D3/D2 preferring], Antagonists: R(+)-SCH 23390 (R(+)-7-chloro-8-hydroxy-3-methyl-1 phenyl-2,3,4, 5-tetrahydro-1H-3-benzazepine), [DA D1-like]; and haloperidol, [DA D2-like]. All drugs were administered intraperitoneally, two injections daily 8 h apart for 30 days. Aromatic L-amino acid decarboxylase (AAAD) and tyrosine hydroxylase (TH) activity, protein and mRNA, as well as DA metabolism were followed with time thereafter in the nigrostriatal neurons. We observed that chronic administration of D1-like agonists had no effect on TH or AAAD activity, while D2-like agonists decreased AAAD, but not TH activity. Additionally, chronic blockade of DA D2-like receptors resulted in prolonged induction of TH and AAAD, while chronic blockade of DA D1-like receptors induced changes of AAAD only. Compared to TH the induction of AAAD was longer lasting. DA metabolism was altered by chronic administration of drugs acting on DA D2-like, but not DA D1 like receptors, and in general the patterns of change did not follow those for TH or AAAD. When studied 48 h after the last dose of the chronic haloperidol schedule TH displayed tolerance to acute drug challenge. At the same time interval, there was tolerance to the enhancing effects of haloperidol and SCH 23390 on DA metabolism. The induction of AAAD by haloperidol or SCH 23990 did not appear to develop tolerance after chronic administration. These observations complement existing knowledge, and provide novel information about AAAD that may have practical importance for Parkinson's patients on L-DOPA therapy. PMID- 10366681 TI - Effects of cyanide and hypoxia on membrane currents in neurones acutely dissociated from the rostral ventrolateral medulla of the rat. AB - Previous reports suggested that some neurones located in the rostral ventrolateral medulla (RVL) can act as fast oxygen sensors which enhance the sympathetic activity and blood pressure independent of peripheral chemoreceptors. The aim of this study was to compare hypoxic responses of different subpopulations of RVL neurones to ascertain whether the hypoxic sensitivity is restricted to one group of these neurones. Whole-cell patch-clamp recordings were made from acutely dissociated neurones obtained from RVL of P13-P19 rats. Short lasting hypoxia (1-2 min) was evoked by pressure injection of NaCN or lowering pO2. Cells projecting to the upper thoracic segments were retrogradely labelled with fluorescent beads. Catecholaminergic (CA) or non-catecholaminergic (non-CA) neurones were identified using single-cell reverse-transcription polymerase chain reaction (RT-PCR) or immunocytochemistry. Recordings were made from 38 neurones (26 spinally-projecting, 12 non-spinal) using Cs+/TEA or K+-containing pipettes. In most of the cells tested with slow depolarising ramp commands (78%; including spinally-projecting and non-spinal neurones, as well as CA and non-CA neurones), NaCN or hypoxia evoked a reversible increase of the sustained inward current. Extracellular application of 1 mM Co2+ or 25 nM TTX revealed three components of the hypoxia-sensitive inward current which resembled the persistent sodium (INaP), low threshold calcium (LVA Ca2+) and high threshold calcium (HVA Ca2+) currents. The NaCN or hypoxia induced increase of the current could also be observed during step commands. Recordings with K+-containing pipettes during similar depolarising ramps revealed, in addition, a reversible increase of IK in 78% of tested cells (in all four types of examined neurones). These results are consistent with the concepts that RVL neurones can act as a central oxygen sensor. However, in contrast to the previously published data demonstrating that in pentobarbital anaesthetised rats only the barosensitive and spinally projecting cells were affected by a short-lasting hypoxia, our findings obtained with dissociated RVL neurones indicate that sensitivity to hypoxia is widely distributed within this part of the medulla oblongata. PMID- 10366682 TI - Ionic mechanism mediating Mytilus inhibitory peptides elicited membrane currents in identified Helix neurons. AB - Effects of seven, pressure applied MIP (Mytilus inhibitory peptides) had been studied on D-neurons of the CNS of Helix pomatia in voltage-clamp experiments. In physiological saline, the peptides produced a hyperpolarization usually coupled with the cessation of any spontaneous spiking activity. Clamped at the resting potential ( approximately -60 mV), peptide applications elicited an outward current, which increased its amplitude by shifting the holding potential towards depolarisation. The response was concentration-dependent and accompanied by an increased membrane conductance. Reversal potentials obtained at different [K+]o were plotted with a slope of 52 mV per ten-fold change in [K+]o showing that the peptide-elicited current was mainly due to the increased K+-conductance(s). The peptide-induced outward current could partially be blocked by Ba2+ (5 mM), CdCl2 (1 mM), TEACl (10 mM) or apamin (2.5x10(-5) M) or furosemide (10 mg/ml) and decreased either in Na+-free or Cl--free solutions. 4-Aminopyridine at 5 mM concentration completely blocked the peptide-induced current. In the presence of high [K+]o, the peptide(s) was still found to induce an outward current at membrane potentials beyond K+-reversal potential. This component was not present in Cl--free saline, suggesting that the current was due to the inward flow of Cl- ions. Our results show that the MIPs have at least two (three) independent actions, each associated with different voltage-, concentration-dependence and ionic mechanisms. It is suggested, that the peptide-induced currents are carried by K+, and Cl- ions. According to our present finding, the observed effects are mediated by the same receptor, activating different second messenger systems, inducing multiple conductance changes in the membrane of neurons of the snail ganglia. PMID- 10366683 TI - Action of capsaicin on dorsal root-evoked synaptic transmission to substantia gelatinosa neurons in adult rat spinal cord slices. AB - An action of capsaicin was investigated on dorsal root-evoked synaptic transmission to substantia gelatinosa (SG) neurons in adult rat spinal cord slices by use of the whole-cell voltage-clamp technique. In 79% of neurons examined, superfusing capsaicin (1 microM) for 30 s depressed a C-fiber-evoked excitatory synaptic current in a manner sensitive to a capsaicin-receptor antagonist, capsazepine (10 microM). On the contrary, Adelta-fiber-evoked excitatory and inhibitory synaptic currents were unaffected by capsaicin in all of cells tested. It is concluded that capsaicin specifically acts on C-afferents, resulting in an inhibition of evoked excitatory transmission to the SG; this may contribute to, at least in part, an acute analgesic action of capsaicin. PMID- 10366684 TI - Columnar activity supports propagation of population bursts in slices of rat entorhinal cortex. AB - Population bursts including epileptiform spikes and sharp waves can be generated in and propagate through the retrohippocampal cortices. The propagation of these events within the entorhinal cortex was studied with field potential recordings in horizontal slices from rat brain. Population bursts were elicited by repetitive extracellular stimuli in normal media. Transections of particular laminae and microknife cuts parallel to the pial surface were used to interrupt propagation or to isolate potential pathways. Population spikes were not found to propagate more than 0.5 mm in either superficial (layers I-III) or deep (layers V VI) strips of tissue formed by cuts along layer IV. Events propagated over 2 mm when 'columnar' connectivity was intact. Population spikes propagated past microknife cuts which started at the pial surface and sectioned layers I-IV. Population spikes also propagated past microknife cuts which sectioned the angular bundle and layers V-VI. When deep layer cuts included layer IV, horizontal propagation of population spikes was blocked. Cuts running approximately in layer IV diminished the amplitude and duration of population events recorded in layers II and V (above and below the cut, respectively) and eliminated a high frequency oscillation which occurred during the burst event. It is concluded that superficial and deep layer neurons interact during propagation of population events in entorhinal cortex and that this 'columnar' activity: (a) intensifies the excitatory activity of superficial and deep layer neurons; and (b) provides multiple paths for the spread of activity from column to column within the entorhinal cortex. PMID- 10366685 TI - Acute in vivo neurotoxicity of peptides from Maedi Visna virus transactivating protein Tat. AB - Lentiviruses such as Maedi Visna virus (MVV) in sheep, and human immunodeficiency virus (HIV) in man often cause a variety of neurological syndromes in later stages of infection. Neuropathological investigations reveal damage to myelin and astrocytosis in both white and grey matter. MVV infection induces axonal damage with some areas of necrosis while neuronal loss, and synaptic damage have been reported in HIV-1 infection. It is not clear, at present, how this neurodegeneration is mediated but, as these viruses do not directly infect neurons, an indirect neurotoxic action of the viruses is indicated. Previous experiments have shown that the intra-striatal injection in rats of a synthetic peptide derived from the basic region of the MVV transactivating protein Tat causes considerable neurotoxicity 1 week post-operatively. By in vivo stereotaxic injections of the same synthetic peptide, and subsequent immunocytochemical detection of neurons, astrocytes and microglia, we show that this neurotoxicity displays a distinctive and unusual lesion profile and is evident as rapidly as 0.5 h post-operatively. Furthermore, neuroprotection studies suggest that the early effects of the MVV tat peptide may involve glutamate neurotoxicity via the N-methyl-D-aspartate (NMDA) receptors since the application of dizolcipine (MK801) reduces the volume of the lesion seen at 1 h after the injection of neurotoxic peptide, while L-NAME is ineffective. The mechanism of this early neurotoxicity is thus different from the longer term actions already described. PMID- 10366686 TI - Oxotremorine-induced modifications of the behavioral and neuroendocrine responses to formalin pain in male rats. AB - In the present investigation, the antinociceptive effects of the muscarinic cholinergic agonist, oxotremorine, were evaluated in rats using the formalin test. In Expt. 1, two oxotremorine concentrations (0.1 and 0.2 mg/kg) and two administration times (15 and 1 min before formalin injection) were chosen. All spontaneous and formalin-evoked behavioral responses were considered. In Expt. 2, only the higher concentration of oxotremorine (0.2 mg/kg) was administered 15 or 1 min before the formalin test. The animals were killed 15, 30 or 60 min after formalin treatment. Blood was collected from the trunk to determine corticosterone plasma levels. Some brain areas (hypothalamus, septum and periaqueductal gray matter) were dissected for determination of the beta endorphin content. Oxotremorine induced a dose- and time-dependent reduction of all formalin-evoked responses: licking was decreased during both the first and second phases of the formalin test, flexing was decreased during the second phase by the higher concentration only and paw-jerk was decreased during the first phase by both concentrations. Rearing and line-crossing were significantly decreased by oxotremorine while exploratory activity was only partially reduced; self-grooming was increased. These effects on exploratory activity and self grooming were abolished by formalin treatment. beta-endorphin content in the septum was increased by oxotremorine administered 15 min, but not 1 min, before formalin-treatment. beta-endorphin in the hypothalamus increased in all formalin treated groups independently of oxotremorine administration. These results confirm, and extend to tonic pain, the analgesic effect exerted by oxotremorine on phasic responses. Because of the different effects on each formalin-induced response, they also indicate both spinal and supraspinal CNS sites of action. PMID- 10366688 TI - Acute interactions between L-DOPA and the neurotoxic effects of 1-methyl-4 phenylpyridinium or 6-hydroxydopamine in mice. AB - We have compared the effects of an i.p. pretreatment with L-DOPA (200 mg/kg) associated with benserazide (25 mg/kg) on neurotoxic effects of either 6 hydroxydopamine (6-OHDA) (50 microg, 10 microl per mouse) or 1-methyl-4 phenylpyridinium (MPP+) (17.5 microg, 10 microl per mouse). The striatal dopamine (DA) content, the vesicular monoamine transporter (VMAT2) density, as well as the hypothalamic norepinephrine (NE) content were measured 8 days after treatments. The L-DOPA-benserazide pretreatment worsened by 65% the 6-OHDA-induced depletion in striatal DA. On the contrary, it reduced by 42% the MPP+-induced depletion in striatal DA and by 54% the MPP+-induced decrease in VMAT2 density. It was noticed that the L-DOPA-benserazide pretreatment did not modify the marked decrease in hypothalamic NE content induced by 6-OHDA. PMID- 10366687 TI - An immunochemical study on tau glycation in paired helical filaments. AB - Glycation is a non-enzymatic posttranslational modification that involves a covalent linkage between a sugar and an amino group of protein molecule forming ketoamine. Subsequent oxidation, fragmentation and/or crosslinking of ketoamine leads to the production of advanced glycation endproducts (AGEs). Formation of AGEs causes detrimental effects on the structure and function of affected proteins. Accumulation of AGEs has been implicated in normal aging and in the pathogenesis of diabetes-associated complications and Alzheimer's disease (AD). Of all AGEs, Nepsilon-(carboxymethyl)lysine (CML) is a major glycoxidation product known to be stable and accumulate progressively in vivo. In order to determine if tau is glycated in AD, we raised a rabbit antibody to CML that demonstrated its usefulness in detecting glycation of different proteins in vitro, including BSA, ribonuclease, lysozyme and recombinant tau. Immunochemical analyses indicated that ribose and glucose-6-phosphate are more effective than glucose in generating CML formation in these proteins. We used this antibody to probe for glycation in the following human tau preparations: tau of normal brains and preparations of soluble PHF-tau as well as insoluble PHF from AD brains. All three principal tau components resolved from PHF-tau on Western blots showed CML immunoreactivity indicating that tau is glycated in PHF-tau; and insoluble PHF exhibited prominent CML immunoreactivity on top of the stacking gel. Moreover, immunoelectron microscopic analyses indicate that the anti-CML antibody labels predominantly PHF in aggregates. Taken together, these results suggest that tau becomes glycated in PHF-tau and glycation may play a role in stabilizing PHF aggregation leading to tangle formation in AD. PMID- 10366689 TI - Interrelation between cerebral energy metabolism and behaviour in a rat model of permanent brain vessel occlusion. AB - The present study investigates the interrelation between cerebral energy metabolism and memory capacities after acute and permanent occlusions of carotid and vertebral arteries in adult Wistar rats (n=60). Tissue ATP, phosphocreatine, ADP, AMP and adenosine concentrations were determined in rat brain by high pressure liquid chromatography (HPLC) analysis. Lactate and pyruvate were measured spectrophotometrically. Rats underwent psychometric testing by means of a holeboard test, closed field activity, and passive avoidance behaviour. Acute cerebral ischaemia was associated with a substantial deficit in energy load ( 50%). Cortical adenosine and lactate exhibited a 7- and a 10-fold increase, respectively, in concentration. After 2 weeks of four-vessel occlusion, cortical ATP and phosphocreatine showed a partial enhancement in their concentrations if compared with acute ischaemia. Consequently, energy load (micromol/g) increased from 0.59 to 1.42 in cerebral cortex and from 0.58 to 1.14 in hippocampus under conditions of acute and permanent ischaemia, respectively. While lactate was normalized, adenosine showed a 2-fold increase in its cortical concentration. All animals improved their abilities in learning, memory and cognition after a 7-day training period. Acute vessel occlusion severely decreased working memory (WM), reference memory (RM) and locomotor activity. Simultaneously, the passive avoidance test showed a significant reduction in latency time from 247+/-85 s (sham) to 145+/-132 s. The partial improvement in brain energy state was accompanied by a relative improvement in WM and RM, although both memory capacities remained significantly lower than in controls. The data of the present study demonstrate a linear relationship between cerebral energy metabolism and brain memory capacities after acute and permanent vessel occlusions in rats. PMID- 10366690 TI - Upregulation of intercellular adhesion molecule-1 expression on human endothelial cells by tumour necrosis factor-alpha in an in vitro model of the blood-brain barrier. AB - Adhesion molecules on the endothelial surface of the blood-brain barrier (BBB) play an important role in the pathogenesis of many encephalopathies, including multiple sclerosis (MS) and cerebral malaria (CM). The expression of four surface molecules of relevance to MS and CM on the immortalized human umbilical vein endothelial cell line, ECV304, was investigated using immunofluorescence flow cytometry. We found that ECV304 cells express intercellular adhesion molecule-1 (ICAM-1) and low levels of CD36, but not vascular cell adhesion molecule-1 (VCAM 1) or E-selectin. This expression pattern was unaltered on ECV304 cells which were co-cultured with C6 glioma cells; conditions under which the endothelial cells display enhanced barrier formation. Tumour necrosis factor-alpha (TNF alpha), which is elevated in MS and CM, decreased the integrity of the barrier in co-cultured endothelial cells and upregulated the expression of ICAM-1 nine-fold. The significance of elevated ICAM-1 expression in relation to the binding of parasitised erythrocytes at the BBB in CM is discussed. PMID- 10366691 TI - Effect of olanzapine on behavioural changes induced by FG 7142 and dizocilpine on active avoidance and plus maze tasks. AB - The present study examined the effect of atypical antipsychotic olanzapine on FG 7142- (N-methyl-beta-carboline-3-carboxamide) and dizocilpine-induced cognitive impairment in active avoidance paradigm and elevated plus maze in mice. Both FG 7142 (5 mg/kg) and dizocilpine (0.1 mg/kg) increased the latency to reach shock free zone (SFZ) both during training and retention session in active avoidance paradigm. This effect was reversed by olanzapine (0.063, 0. 125, 0.25 and 0.5 mg/kg). Similarly, FG 7142 (5 mg/kg) increased transfer latency (TL) on both first and second day in elevated plus maze. The lower doses of olanzapine (0.063 and 0.125 mg/kg) reversed the effect of FG 7142 on second day in elevated plus maze but higher doses (0.25 and 0.5 mg/kg) failed to modify the effect of FG 7142 both on first and second day. Dizocilpine (0.1 mg/kg) treatment did not affect TL on first day while on second day, it increased TL significantly. Olanzapine (0.063 and 0.125 mg/kg) reversed the effect of dizocilpine on elevated plus maze but the higher doses (0. 25 and 0.5 mg/kg) failed to reverse it. Even though olanzapine (0. 063, 0.125 and 0.25 mg/kg) failed show any effect per se in active avoidance task, the higher dose (0.5 mg/kg) increased the latency to reach SFZ on second day. Olanzapine (0.063, 0.125, 0.25 and 0.5 mg/kg) did not show any per se effect on TL in elevated plus maze on first day while on second day, olanzapine (0.125, 0.25 and 0.5 mg/kg) increased TL as compared to control group. The present study demonstrated olanzapine's reversal of dizocilpine- and FG 7142 induced behavioural changes in active avoidance paradigm and elevated plus maze. Although the precise mechanism of action is unknown, olanzapine might be acting by blocking excessive dopaminergic activity in the prefrontal cortex. PMID- 10366692 TI - Subpopulations of neurons in the rat neostriatum display GABABR1 receptor immunoreactivity. AB - Immunoreactivity for gamma aminobutyric acid BR1 receptor (GABABR1) was detected in the neuropilar elements as well as in the perikarya of neurons in the neostriatum. Many of the GABABR1-immunoreactive perikarya were medium-sized with a thin rim of cytoplasm. They resembled the morphology of medium spiny neurons, the projection neurons of the neostriatum. In addition, some GABABR1 immunoreactive neurons were densely labeled and were of medium to large in size. These neurons were characterized by double immunofluorescence using their neurochemicals as markers. Over 90% of the parvalbumin- and choline acetyltransferase-immunoreactive neurons and about 80% of the nitric oxide synthase-immunoreactive neurons displayed GABABR1 immunoreactivity. The present results show for the first time that the major four subpopulations of striatal neurons express GABABR1 receptor and may have a functional implication in the GABA neurotransmission in the microcircuitry of the neostriatum. PMID- 10366693 TI - Different spatial distributions of sodium channels in the slowly and rapidly adapting stretch receptor neuron of the crayfish. AB - Inward Na+ currents were studied, using a two-microelectrode intracellular voltage-clamp technique, in the slowly adapting (SA) and rapidly adapting (RA) stretch receptor neurons of the crayfish after the axons were cut at different distances from the soma. In the SA neuron, inward Na+ currents were recorded in the soma even when the axon was cut as close as 100 microm from the center of the soma, indicating the presence of Na+ channels in these parts. Also, two populations of Na+ channels seem to exist in the SA neuron. In the RA neuron, only minute Na+ currents were observed if the axon was shorter than 250 microm. The results strongly indicate that the voltage-gated Na+ channels in the SA and RA neurons have different distributions and that the difference in the spatial distribution of Na+ channel types may be important for the difference in firing properties in the two types of neurons. PMID- 10366694 TI - Expression of intracellular progesterone receptors in rat brain during different reproductive states, and involvement in maternal behavior. AB - Progesterone is one of a complex of hormones which influences the occurrence of maternal behavior in rats. The present study provides information on progesterone's mechanism and possible neural site(s) of action with respect to maternal responsiveness. Progesterone can exert cellular effects by acting on membrane receptors or by acting on intracellular receptors. In the first experiment we show that RU 486 can antagonize progesterone's inhibitory effect on maternal behavior. Since RU 486 acts as an antagonist to progesterone's action at its intracellular receptor, these results support the involvement of that receptor in maternal behavior control. The second experiment employs immunocytochemical techniques to detect the number of cells in various forebrain regions which contain intracellular progesterone receptors during different reproductive states. The number of cells which contained progesterone receptors was higher toward the end of pregnancy (progesterone is presumably exerting its effects on maternal behavior at this time) when compared to either early pregnancy or lactation in the following forebrain regions: anteroventral periventricular nucleus of the preoptic area; medial preoptic area; ventral part of the bed nucleus of stria terminalis; ventrolateral division of the ventromedial nucleus; arcuate nucleus; anterior paraventricular nucleus of the hypothalamus; and medial amygdala. The possible involvement of these regions as a site or sites where progesterone might exert its effects on maternal behavior is discussed. PMID- 10366695 TI - Occurrence and distribution of substance P receptors in the cerebral blood vessels of the rat. AB - The distribution of immunoreactivity to the receptor for substance P was examined in the cerebral blood vessels of the rat. Substance P immunoreactivity has been demonstrated in the nerve fibers of the cerebral blood vessels. Recently, the production of substance P receptor specific antibody has enabled the detection of localization of the substance P receptor in the central nervous system. In this study, we examined the existence of nerve fibers with substance P receptor immunoreactivity in the cerebral blood vessels and the cranial ganglia innervating the cerebral blood vessels. Sprague-Dawley rats were perfused with fixative and the pial arteries and the cranial ganglia known to innervate the cerebral blood vessels, i.e., trigeminal, sphenopalatine, internal carotid, otic and superior cervical ganglia, were dissected. All specimens were incubated with anti-substance P receptor IgG, then stained by the avidin-biotin-peroxidase complex method. Numerous nerve fibers with varicosities forming plexuses, with substance P receptor immunoreactivity were observed on the walls of the major extracerebral arteries forming the circle of Willis and its branches. Substance P receptor immunoreactivity was also detected in the endothelium of the cerebral arteries. Substance P receptor immunoreactivity was positive in many neurons of the sphenopalatine ganglion, otic ganglion, trigeminal ganglion, superior cervical ganglion and internal carotid ganglion. The present study demonstrated the existence of nerve fibers with substance P receptor immunoreactivity in the cerebral blood vessels and the cranial ganglia that innervate the cerebral blood vessels. These findings are important in understanding the responsiveness of the cerebral blood vessels to substance P. PMID- 10366696 TI - GABA induces proliferation of immature cerebellar granule cells grown in vitro. AB - The presence of GABA and its receptors early in rodent nervous system development has lead to speculation on the role of this transmitter system in neuroblast proliferation, migration and differentiation. We studied the effect of GABA and GABA agonists on immature cerebellar granule cell proliferation and survival. Cerebellar granule cell suspensions were obtained from 6-8-day-old rats and grown in culture for up to 7 days in serum-containing or serum-free medium. The addition of GABA (0.1-100 microM) or muscimol (0.01-10 microM) 2 h after inoculation and harvested 22 h later, lead to an increase in 3H-thymidine incorporation over control samples with the correspondent increase in granule cells number assayed 48 h later. The effect on cell proliferation exerted by GABAA agonists was blocked by MgCl2 and nifedipine, as well as by the chloride channel blocker, picrotoxin (50 microM), and the GABAA receptor specific blocker, bicuculline (50 microM). The increase on cell proliferation induced by GABA also was blocked by PD98059 (75 microM), a specific inhibitor of the mitogen-activated protein kinase kinase (MAPKK). GABAA receptor-mediated proliferation was consistently seen in cells inoculated in serum-containing medium supplemented with 25 mM KCl but not seen in serum-free medium, with 5 mM or 25 mM KCl. The presence of serum did not enhance the survival of cerebellar granule cells grown for 7 days in either 5 mM or 25 mM KCl. Additionally, neither GABA nor muscimol applied from day 2 to day 7 in vitro affected cell survival in any culture condition. We conclude that GABA and GABAA receptor agonists influence granule cell proliferation but not survival and that this effect is mediated by a calcium influx via voltage-dependent calcium channel activation, with a subsequent activation of the MAPK cascade. PMID- 10366697 TI - High sensitivity of immature GABAergic neurons to blockers of voltage-gated calcium channels. AB - The involvement of voltage-gated calcium channels in the survival of immature CNS neurons was studied in aggregating brain cell cultures by examining cell type specific effects of various channel blockers. Nifedipine (10 microM), a specific blocker of L-type calcium channels, caused a pronounced and irreversible decrease of glutamic acid decarboxylase activity, whereas the activity of choline acetyltransferase was significantly less affected. Flunarizine (1-10 microM, a relatively unspecific ion channel blocker) elicited similar effects, that were attenuated by NMDA. The glia-specific marker enzymes, glutamine synthetase and 2',3'-cyclic nucleotide 3'-phosphohydrolase, were affected only after treatment with high concentrations of nifedipine (50 microM) or NiCl2 (100 microM, shown to block T-type calcium channels). Nifedipine (50 microM), NiCl2 (100 microM), and flunarizine (5 microM) also caused a significant increase in the soluble nucleosome concentration, indicating increased apoptotic cell death. This effect was prevented by cycloheximide (1 microM). Furthermore, the combined treatment with calcicludine (10 nM, blocking L-type calcium channels) and funnel-web spider toxin-3.3 (100 nM, blocking T-type channels) also caused a significant increase in free nucleosomes as well as a decrease in glutamic acid decarboxylase activity. In contrast, cell viability was not affected by peptide blockers specific for N-, P-, and/or Q-type calcium channels. Highly differentiated cultures showed diminished susceptibility to nifedipine and flunarizine. The present data suggest that the survival of immature neurons, and particularly that of immature GABAergic neurons, requires the sustained entry of Ca2+ through voltage-gated calcium channels. PMID- 10366698 TI - Endogenous levels of 5 alpha-reduced progestins and androgens in fetal vs. adult rat brains. AB - 5 Alpha-reduced metabolites of certain steroids have been shown to have important functions in adult brains and may play a role in brain development. To assess which 5 alpha-reduced steroid metabolites may have an impact during development, endogenous levels of 5 alpha-reduced androgens and progestins and their parent hormones were measured in male and female fetal brains over the last 5 days of gestation. These levels were compared to levels measured in adult male and female brains (evaluated at different stages of the estrous cycle). Neither the brain levels of parent hormones nor of their 5 alpha-reduced metabolites varied as a function of fetal sex or of gestational age. Therefore, the data from the two sexes were combined. In fetal brains, the levels of the progesterone reduced metabolites were 20-fold higher than levels of progesterone itself whereas levels of testosterone reduced metabolites were 10-fold lower than testosterone levels. In contrast to fetal brain, conversion of progesterone to reduced metabolites was much lower in adult brain, but the level of 5 alpha-reduced androgens was 3-10 fold higher than the level of testosterone in all adult tissue, indicating more conversion of androgen to 5 alpha-reduced metabolites in adult than in fetal brains. These results imply that the reduction of progesterone to reduced metabolites may play a critical role in brain development. PMID- 10366699 TI - Synergistic effects of brain-derived neurotrophic factor and ciliary neurotrophic factor on cultured basal forebrain cholinergic neurons from postnatal 2-week-old rats. AB - Brain-derived neurotrophic factor (BDNF), a member of the neurotrophin family, and ciliary neurotrophic factor (CNTF), a member of the neurocytokine family, are known to have synergistic effects on motoneurons, but such synergistic effect has not been studied in detail especially in the brain. In the present study, we examined the synergistic effects of BDNF and CNTF on the survival of basal forebrain cholinergic neurons cultured from postnatal 2-week-old (P2w) rats. Although BDNF is well-known to promote the survival of basal forebrain cholinergic neurons in P2w culture, CNTF had little effect on the survival of choline acetyltransferase (ChAT)-positive neurons and did not increase ChAT activity in the culture. However, CNTF enhanced BDNF-mediated promotion of cell survival of cholinergic neurons when added concomitantly. BDNF alone induced only a three-fold increase in ChAT activity in control cultures, but the concomitant addition of CNTF resulted in an eight-fold increase. CNTF did not enhance BDNF mediated cell survival of total neurons from the basal forebrain, hippocampus or cerebellum, suggesting that the synergistic effects of CNTF on the BDNF-mediated increase of viability might be strong in basal forebrain cholinergic neurons. CNTF also enhanced the neurotrophin-4/5-mediated increase of ChAT activity, but not the nerve growth factor (NGF)-mediated one. Furthermore, the BDNF-mediated increase was also enhanced by leukemia inhibitory factor but not by interleukin 6. Similar synergistic pattern between neurotrophins and cytokines were also observed in the induction of ChAT activity in embryonic basal forebrain culture. These results suggest that TrkB, a functional high-affinity receptor of BDNF and NT-4/5, and LIFR beta, a receptor component contained in CNTF and LIF receptor complex, might be involved in the observed synergistic effects. PMID- 10366700 TI - The NMDA antagonist, MK-801, alters L-DOPA-induced air-stepping in neonatal rats. AB - Administration of L-DOPA (sc) to neonatal rats suspended in harnesses induces coordinated stepping of all four limbs (diagonal progression; L-DOPA-induced air stepping) by 5 days of age. Because NMDA also induces locomotion in several species, NMDA receptor activation may be required for L-DOPA to elicit coordinated air-stepping. The purpose of the present experiment was to determine if the NMDA receptor antagonist, MK-801, would block L-DOPA-induced air-stepping in developing rats. Neonatal rats administered MK-801 alone rarely air-stepped with the forelimbs or hindlimbs in a coordinated fashion, whereas those treated with L-DOPA alone primarily stepped with all four limbs using a diagonal progression pattern during the session. In contrast, the number of limbs that stepped during the session was gradually altered in 5- to 20-day-old rats treated with MK-801 + L-DOPA. Gaits of those rats progressed from diagonal progression to extension of the forelimbs beneath the chin with hindlimb alternation, to forelimb extension without hindlimb activity. Twenty-day-olds treated with MK-801 + L-DOPA subsequently became completely inactive when the forelimbs dropped from their elevated position beneath the chin. In addition to the sequence just described, 15-day-old rats treated with the lowest concentration of MK-801 + L DOPA occasionally stepped with one pair of homolateral limbs or stepped with the hindlimbs in near synchrony while the forelimbs either stepped in alternation, were extended beneath the chin or groomed the face. Because limb participation during L-DOPA-induced air-stepping was altered in neonatal rats pretreated with MK-801, NMDA receptor activation may be important for locomotor coordination (gait). PMID- 10366701 TI - Characterization of the cell death promoter, Bad, in the developing rat retina and forebrain. AB - Neuronal programmed cell death, or apoptosis, occurs during development, following injury or in certain disease processes, and is regulated by members of the B-cell leukemia-2 (Bcl-2) protein family. These molecules include both positive and negative regulators of cell death and act by selective dimerization that results in permissive or inhibitory effects on a cascade of cellular events, including mitochondrial release of cytochrome c, stimulation of cysteine protease activity and subsequent cellular deterioration. Here, we have characterized the expression of the cell death agonist, Bad, in the postnatal rat retina and forebrain. Isolation, subsequent amplification by RT-PCR and DNA sequence analysis revealed that retinal Bad was identical to Bad expressed in the developing and adult rat brain. Using a polyclonal antibody to Bad, we determined that, in the retina, on the day of birth (postnatal day-0, PND-0) Bad immunoreactivity was expressed primarily by retinal ganglion cells, some cells in the inner neuroblastic layer (NBL) and an indistinct plexus of processes in the inner plexiform layer (IPL). On PND-7, Bad immunoreactivity was observed in most cells in the ganglion cell layer (GCL), numerous cells scattered throughout the inner nuclear layer (INL), a lightly stained IPL and in a distinct band of immunostained fibers in the forming outer plexiform layer (OPL). By PND-15, Bad immunoreactivity was present in cells in the GCL, in some cells in the proximal INL and in horizontal cell processes in the OPL. The IPL was only faintly labeled. In the adult retina, specific Bad immunostaining was confined to large cells in the ganglion cell layer (presumed ganglion cells), occasional lightly stained horizontal cells and their processes in the OPL and to occasional small, lightly stained cells in the proximal INL (presumed amacrine cells) and GCL (presumed displaced amacrine cells). Again, the interposed IPL was faintly labeled. In the brain, Bad immunoreactive cells were scattered throughout the forebrain parenchyma but were particularly concentrated in neurons of the cerebral cortex, hippocampus and amygdala. Bad immunoreactivity was heaviest in these cells at PND-7, distinctly weaker at PND-10 and absent by PND-24. At all time points examined, Bad immunoreactivity was present in epithelial cells of the choroid plexus, as previously reported in the adult rat brain. These data suggest that Bad is transiently expressed by various cell types in the perinatal retina, particularly ganglion cells, and in discrete forebrain regions. In the context of corroborative observations, Bad expression may be regulated in response to acute ischemia and may act as a control point for retinal neuronal apoptosis. PMID- 10366702 TI - Apoptosis in the development of the mouse olfactory epithelium. AB - Apoptotic cells were detected in the mouse olfactory epithelium (OE) at different embryonic and postnatal stages by in situ nick translation (ISNT) and Tdt mediated dUTP nick end-labeling (TUNEL) techniques. During development, the apoptotic process presented two peaks. One at E12 during the invagination of the olfactory placode and the second at E16 corresponding to olfactory axon synaptogenesis. Then, from E18, a sharp decrease in the number of apoptotic cells was observed and at E19 the apoptotic index reached low values that were maintained in postnatal stages, P1 and P8, and in the adult. Apoptotic nuclei belonged to mature as well as immature olfactory receptor neurons (ORNs). Indeed, double-labeling experiments evidenced apoptotic neurons immunopositive for olfactory marker protein (OMP), carnosine and GAP-43. According to our data, two apoptotic phases occur during early development. One is involved in the morphogenesis of the OE when this last is not yet, or poorly, connected to its target, the olfactory bulb (OB). The second peak of apoptosis is more closely dependent on the interplay between OE and OB. PMID- 10366703 TI - Regulated expression of estrogen receptor alpha and beta mRNA in granule cells during development of the rat cerebellum. AB - A semi-quantitative RT-PCR approach was used to characterize expression of the mRNA encoding estrogen receptors in developing cerebellar granule cells of the rat. Evidence is presented for expression of both ERalpha and ERbeta transcripts in granule cells throughout the first 15 postnatal days. While transcripts encoding both ERalpha and ERbeta were expressed in granule cells, the relative levels of expression varied significantly during the first two postnatal weeks of cerebellar development. The ERalpha mRNA was expressed at the lowest level on the first day following birth; whereas expression of ERbeta was highest on that day. On the fourth postnatal day the expression of ERalpha increased, while there was a significant decrease in ERbeta expression. Between postnatal day 4 and 15, the expression of the mRNA of each receptor varied in a similar fashion; expression decreased slightly between days 4 and 10 and then increased significantly on day 15. Alternative splicing of the ERbeta transcripts was also investigated and was likewise found to vary during granule cell development. Initially, the mRNA encoding the beta1 isoform was predominant, but by day 4, the beta2 isoform was the major isoform expressed. On postnatal days 7 and 10, there was not a significant difference between the level of beta1 and beta2 expressed. By day 15, beta1 was again the predominant ERbeta isoform accounting for nearly 90% of all ERbeta transcripts expressed. PMID- 10366704 TI - Microglia in the human fetal spinal cord--patterns of distribution, morphology and phenotype. AB - Microglia, the intrinsic macrophages of the nervous system, colonise the cerebrum around the second trimester in man. In order to determine the extent of microglial influx into the nervous system, we have examined their distribution within the human fetal spinal cord in relation to astrocytic and vascular development between 9 and 16 weeks of gestation, using conventional immunohistochemistry [CD11b; CD45; CD64; CD68; ICAM-1; ICAM-2; VCAM-1; PECAM; GFAP; vimentin] and lectin histochemistry [RCA-1]. Microglia are identifiable by 9 weeks, within the ventricular/sub-ventricular zones. Human fetal microglia display heterogeneity in phenotype and are more readily identified by CD68 in the spinal cord. There is a marked influx of cells dorsal and ventral to the neural cavity, from the marginal layer [meninges/connective tissue] with advancing gestational age, with greatest cell densities towards the end of the time period in this study. This inward migration is associated with progressive vascularisation, ICAM-2 expression and co-localises with GFAP and vimentin positive radial glia. The patterns of microglial migration in human fetal cord differ from that within the cerebrum, but generally conform to a route following white to gray matter. PMID- 10366705 TI - Postnatal development of detergent-insoluble properties of NMDA and AMPA receptor subunits in the rat brain synaptic membrane. AB - We investigated the developmental changes of detergent-insoluble characteristics of NMDA and AMPA receptor subunits in the synaptic membranes prepared from the rat cerebral cortex. At postnatal day (PND) 1, the majority of NMDAR1 and NMDAR2B subunits of NMDA receptors in the synaptic membranes were insoluble to the treatment of 1% Triton X-100. The detergent-insoluble properties of both subunits were not significantly changed during postnatal development. At PND 1, about 45% of GluR1 and 10% of GluR2/3 subunits of AMPA receptors in the synaptic membrane were insoluble to Triton X-100, whereas 70% of GluR1 and 56% of GluR2/3 subunits were insoluble at PND 22. These findings indicate that the postsynaptic clustering of NMDA and AMPA receptors during development seems to be differentially regulated in vivo. PMID- 10366706 TI - Differential expression of Islet-1 in neural crest-derived ganglia: Islet-1 + dorsal root ganglion cells are post-mitotic and Islet-1 + sympathetic ganglion cells are still cycling. AB - The Islet-1 antigen is an early marker of differentiation of neural tube cells, and is expressed in many other embryonic cells as well. It had been reported that Islet-1 is expressed only in post-mitotic sympathetic neuroblasts in vitro, unlike other differentiation markers. We have double-labeled St. 23 chick embryos for bromodeoxyuridine (BrDU) and Islet-1 and found that neural tube and dorsal root ganglion (DRG) cells express Islet-1 after leaving the cell cycle, while sympathetic ganglion (SG) cells express Islet-1 while still dividing. PMID- 10366707 TI - Transient expression of calcitonin gene-related peptide immunoreactivity in the ventral horn of the post-natal rat cervical spinal cord. AB - In addition to the well known expression of calcitonin gene-related peptide (CGRP) immunoreactivity in primary afferent fibers in the dorsal horn and in motoneurons, this study has demonstrated, in rat, transient CGRP immunoreactivity in fine caliber varicose axons throughout the ventral horn and in a group of neuron cell bodies in the medial ventral horn. This was first observed at post natal day 7 (P7) and had disappeared by P21. Physiological studies in chick embryonic spinal cord have shown that CGRP modulates spontaneous activity during development [Carr, P.A., Wenner, P., 1998. Calcitonin gene-related peptide and effects on spontaneous activity in embryonic chick spinal cord. Dev. Brain Res. 106, 47-55]. Neural activity increases post-natally in rat where it may play a role in refinement of sensorimotor synapses. This activity may also be modulated by CGRP. PMID- 10366708 TI - Connexin43 mRNA and protein expression during postnatal development of defined brain regions. AB - Connexin43 (cx43) mRNA and protein were quantified in seven brain regions using RNase protection assays and Western blotting from postnatal day 3, 10, and 17, and adult Sprague-Dawley rats. Increases in cx43 mRNA abundance preceded strong increases in cx43 protein in the cerebral cortex, hippocampus, hypothalamus, and striatum, whereas smaller postnatal increases were seen in the cerebellum, brain stem, and olfactory bulbs. PMID- 10366710 TI - Organisation of the mouse and human 5T4 oncofoetal leucine-rich glycoprotein genes and expression in foetal and adult murine tissues. AB - The human 5T4 oncotrophoblast leucine-rich glycoprotein may contribute to the process of placentation or metastasis by modulating cell adhesion, shape and motility. To understand better the role of 5T4 in development and cancer, the gene structure has been elucidated from both human and mouse genomic clones and mRNA expression has been studied in foetal and adult mouse tissues. The protein coding region is located within the second of two exons, the first exon comprising solely of 5'-untranslated region. Upstream there are no TATA or CAAT boxes, but there are a number of potential Sp1 binding sites. The murine and human proteins show a homologous domain organisation of the leucine rich repeats (LRR) and associated N- and C-terminal flanking regions, although the hydrophilic sequence which intervenes between the two LRR domains contains six additional amino acids in the mouse. The signal peptide, transmembrane region and cytoplasmic tail sequences are identical as are 6 out of the 7 potential N-linked glycosylation sites. Mouse 5T4 transcripts are abundant in placenta and also highly expressed in embryos while in adult tissues transcripts are restricted to brain and ovary. These patterns of expression and the genomic organisation are discussed in relation to possible function and other recently described LRR containing proteins. PMID- 10366709 TI - Protein-DNA footprinting of the human epsilon-globin promoter in human intact cells using nitrogen mustard analogues and other DNA-damaging agents. AB - Nitrogen mustard analogues, bleomycin and dimethyl sulphate (DMS) have been used as probes of protein-DNA interactions in intact human cells. The sites of damage have been determined at base pair resolution in the single copy epsilon-globin gene promoter in erythroid K562 cells, non-erythroid HeLa cells and purified DNA. Exponential amplification of gene-specific damage fragments was achieved using the ligation-mediated polymerase chain reaction (LMPCR) technique and analysed on DNA sequencing gels. A comparison of the relative damage band intensities between purified DNA and intact cells revealed several significant differences - both protection (footprint) and enhancement. These differences occurred at putative transcription factor binding sites and hence are thought to be due to protein-DNA interactions. A major feature of the band intensity ratio plots was the footprint observed at the CCAAT box binding motif as revealed by nitrogen mustard analogues. Enhanced band intensity (hypersensitivity) was displayed at the 5'- and 3'-ends of the CCAAT box in K562 cells - this feature was absent in HeLa cells and in vitro reconstitutions. A footprint was found at the GATA-1 motif in K562 cells that was also absent in non-expressing HeLa cells. Footprints were also evident at the TATA box, CACC box and the epsilonF1 DNA binding motif in K562 cells. PMID- 10366711 TI - Isolation and characterization of the gene coding for Artemia franciscana TATA binding protein: expression in cryptobiotic and developing embryos. AB - Genomic and cDNA clones coding for the Artemia franciscana homolog of the TATA box-binding protein (TBP) were isolated. The C-terminal region of the predicted protein displays up to 92% sequence identity with the conserved C-terminal regions of TBPs from other species. The gene is divided in seven exons that expand over a region of 33 kb. The position of the four introns located in the conserved C-terminal region has been compared with those of other species. Two of these introns have been generally conserved during evolution, another is an arthropod specific intron, present in Drosophila melanogaster and A. franciscana, and the other is only conserved between vertebrates and A. franciscana. Primer extension experiments detected several transcription initiation sites. Northern blot analyses showed the presence of four mRNAs of estimated sizes of 6.8, 2.6, 1.6 and 1.1 kb. Except for the low expression of the 6.8 and 2. 6 kb RNAs in encysted embryos, steady-state levels showed little variation during the activation of the encysted embryo and the first steps of embryonic and larval development. The amount of TBP protein expressed in encysted embryos and developing larvae has been analyzed by Western blot. Cryptobiotic embryos contain significant amounts of TBP although the level of expression increased almost twice during the first 20 h of development. The presence of TBP protein in cryptobiotic embryos suggests that TBP does not play, by itself, a critical role in the arrest of transcription characteristic of these resistance forms. PMID- 10366712 TI - Schistosoma TOR (trispanning orphan receptor), a novel, antigenic surface receptor of the blood-dwelling, Schistosoma parasite. AB - Sh-TOR is a novel, putative three transmembrane domain receptor of Schistosoma haematobium, which has no extensive homology to any other known protein. The 0.86 kb open reading frame was found to encode a novel protein, 286 amino acids long and of 32 kDa. It was shown that Sh-TOR can be phosphorylated on tyrosine and the protein sequence reveals a long cytoplasmic tail with several consensus phosphorylation sites for enzymes which characteristically associate with membrane receptors. The proposed topology of Sh-TOR, based on antibody recognition of transfected Sh-TOR, predicts that the amino terminus is extracellular and the carboxyl terminus intracellular. Sh-TOR is a non glycosylated protein found in the surface tegumental plasma membrane, and tegumental surface pits of adult schistosomes. The 1.35 kb transcript was most highly expressed in the larval stage, which is more susceptible to immune attack. A TOR homologue from Schistosoma mansoni is also described. A homologue from Trypanosoma cruzi, another human parasite was also isolated, but not from the free-living nematode Caenorhabditis elegans. Recombinant Sh-TOR is specifically recognised by a passively protective serum, from baboons vaccinated with irradiated Schistosoma parasite. Together with its surface location, this means that Sh-TOR is also a potential vaccine candidate molecule. PMID- 10366713 TI - Characterization of a kidney-specific pattern of chromatin structure in the rat phosphoenolpyruvate carboxykinase gene. AB - The kidney-specific chromatin structure of the phosphoenolpyruvate carboxykinase (PEPCK) gene was examined and compared to that of the liver. Kidney nuclear extracts were found to lack a liver-enriched factor, pepA, that binds to HSS A, a distal enhancer of the PEPCK gene that may be involved in opening the chromatin domain of the PEPCK gene in the liver. To begin the characterization of the kidney-specific chromatin structure of the PEPCK gene, nuclease hypersensitive sites (HSS) were mapped by indirect end-labeling analysis in proximal tubules from control rats, proximal tubules from acidotic rats which express induced levels of PEPCK, and NRK52E cells, a rat kidney epithelial cell line which does not express the PEPCK gene. A subset of HSS, at -400/+1 over the proximal promoter and at +1900 within the coding region, correlate with kidney-specific PEPCK expression. Two other HSS, at -3.1 kb and +6.2 kb, are detected in kidney cells regardless of PEPCK expression. The HSS at -4800, -1240, and +4650, previously identified in PEPCK-expressing liver cells, were not observed in the kidney. As in the liver, the pattern of hypersensitivity in the kidney does not change by altering the rate of transcription. PMID- 10366714 TI - Transcriptional regulation of the human ADP-ribosylation factor 5 (ARF5) gene. AB - Hybridization of a blot containing 50 human RNAs with an ADP-ribosylation factor 5-specific (ARF5) oligonucleotide probe revealed that the ARF5 gene is expressed in all tissues; however, the level of expression varies significantly with highest levels in pancreas, pituitary gland, and placenta. The 5'-flanking region of the human ARF5 gene lacks a TATA or CAAT box and is highly GC-rich. Primer extension analysis indicates that transcription initiates at a discrete site 62 bp 5' to the start of translation; however, the sequence surrounding the transcription initiation site does not resemble the initiator elements described for other TATA-less genes. Transient transfection of ARF5/luciferase deletion constructs into human IMR-32 neuroblastoma cells revealed that sequences within 169 bp of the transcription initiation site were necessary for full expression. Two GC boxes within this region were modified by site-directed mutagenesis and found to be critical for expression of the reporter constructs. Electrophoretic mobility-shift assays demonstrated specific DNA/protein complexes could be formed with oligonucleotides containing each of the GC boxes and these complexes could be effectively competed by oligonucleotides containing either ARF5 Sp1 site or by an oligonucleotide containing a previously characterized Sp1-binding sequence. The level of ARF5 gene expression, therefore, is dependent upon Sp1 or an Sp1 like factor but does not rely upon a canonical initiator element for accurate transcription initiation. PMID- 10366715 TI - Structural and functional analysis of the rat metallothionein III genomic locus. AB - Metallothionein III (MT III) has been reported to suppress neuronal growth in a rat in vitro model system. The protein and its specific mRNA are detected predominantly in the brain, differentiating MT III from the well-characterised archetypal metallothioneins. Isolation, sequencing and functional analysis of the rat MT III genomic locus indicated that, although the organisation of the gene was conserved between MT III and the more conventional metallothioneins, the 5' flanking region of the MT III gene was distinct. Within this region, a number of putative regulatory elements were identified, including the metal regulatory elements (MREs) characteristic of metallothionein promoters. However, despite their conservation in sequence with active elements, the MREs of MT III were unresponsive to zinc. A 'silencing element' was revealed within a 250 bp section of the MT III promoter which suppressed gene expression in two brain cell lines. The operation of this silencing region in conjunction with the inactive MREs may explain the distinct expression profile observed for MT III within the central nervous system and during neuronal development. PMID- 10366716 TI - Promoter analysis of the human centromeric and telomeric survival motor neuron genes (SMNC and SMNT). AB - Proximal spinal muscular atrophy (SMA) is caused by mutations in the telomeric (SMNT), but not centromeric (SMNC), survival motor neuron gene. Here we have identified and analyzed the two SMN promoters. We show that a 750-bp 5'-flanking fragment from each is capable of driving expression from a reporter construct. Within this fragment, we define a approximately 200-bp element that results in high expression in a motor neuron cell line. Sequence comparison of a 3. 4-kb upstream fragment from each gene shows minimal differences. Although these differences produce a 2-fold difference in reporter activity between the two promoters, this is not sufficiently high to explain why SMNT, but not SMNC, is the disease determining gene. Our data thus demonstrate, for the first time, almost complete equivalence between the SMN promoters and rule out the important possibility that differences in them might explain why mutations in only the telomeric SMN gene cause SMA. PMID- 10366717 TI - Identification of a 24 kDa intrinsic membrane protein from mammalian peroxisomes. AB - A 24 kDa protein from rat liver peroxisomal membrane was isolated and subjected to Edman degradation. Using the N-terminal sequence of this polypeptide we have identified several rat and human expressed sequence tags in the GenBank Database. The complete sequence of a human cDNA clone was determined. The open reading frame encodes an extremely basic protein 212 amino acid residues long. A high similarity between this mammalian protein and hypothetical proteins from Caenorhabditis elegans and Neurospora crassa was found. Hydropathy analysis reveals the existence of two putative membrane-spanning domains in conserved regions of the three homologous proteins. PMID- 10366718 TI - The mRNA-binding ribosomal protein S26 as a molecular marker in plants: molecular cloning, sequencing and differential gene expression during environmental stress. AB - One condition for using a gene as a transcriptional marker for environmental stress is its specific and differential expression. In order to be used as such a marker, the ribosomal protein S26 cDNA from pea (Pisum sativum L.) was cloned and fully sequenced. The gene (PsRPS26) was shown to be differentially regulated by ozone and UV-B radiation in opposite ways. Ozone gave rise to increased mRNA levels, whereas UV-B led to a decrease in S26 transcript abundance. Thus, the expression of PsRPS26 can be used as a molecular marker to differentiate between these two environmental stresses. PMID- 10366719 TI - Analysis of the mouse MAP1B gene identifies a highly conserved 4.3 kb 3' untranslated region and provides evidence against the proposed structure of DBI-1 cDNA. AB - We determined the previously unknown 3' end of MAP1B mRNA. We found an unusually long and highly conserved 3' untranslated region (3'UTR) of 4.3 kb and detected a polymorphism in the 3' flanking region probably due to the integration of an endogenous retroviral MuERV-L element. Furthermore, we found that MAP1B 3'UTR overlapped with the 5' end of the cDNA encoding DBI-1. However, further analysis suggested that the published structure of DBI-1 cDNA is most likely the result of fortuitous joining of unrelated cDNA fragments during cloning. PMID- 10366720 TI - Identification of cdc2cAt: a new cyclin-dependent kinase expressed in Arabidopsis thaliana flowers. AB - In the present paper, the isolation of a third cyclin-dependent kinase gene and its cognate cDNA from Arabidopsis thaliana is described. Whereas other characterised cdc2 genes are ubiquitously expressed in Arabidopsis, expression of cdc2cAt is restricted to flowers. This gene, named cdc2cAt, differs from the two previously reported cdc2 genes in its organisation. Comparison with other cdc2 genes suggests that the deduced protein belongs to a new family of CDC2-like proteins related to the human CHED protein kinase. PMID- 10366721 TI - Cloning and characterisation of the gene for the large subunit of the DNA primase from Drosophila melanogaster. AB - We have cloned the gene for the large subunit of the DNA primase from Drosophila melanogaster, and mapped it to position 77b on chromosome 3. The central region of the protein shows high similarity with homologues from other species, but the N- and C-termini diverge. The protein is enriched in replicating tissues, and consistent with this the region upstream of the gene contains close matches to the sites of two transcription factors - Dref and E2f - which have been implicated in controlling proliferation-associated genes. PMID- 10366722 TI - cDNA and structural organization of the gene Pole1 for the mouse DNA polymerase epsilon catalytic subunit. AB - The cDNA and the gene for the mouse DNA polymerase epsilon catalytic subunit were cloned. The deduced protein sequence shows remarkable evolutionary conservation in DNA polymerase epsilon family. However, several conserved elements involved in template-primer binding differ from those of other class B polymerases. This is likely to reflect a distinctive function of the enzyme. The gene that was assigned to chromosome 5 region E3-E5, consists of 49 exons and has a non conforming splice site in the junction of exon and intron 13. A CpG island covers the promoter region which contains several putative consensus elements critical for S phase upregulated and serum responsive promoters. PMID- 10366723 TI - Reexamination of the high mobility group-1 protein for self-association and characterization of hydrodynamic properties. AB - Previous studies of the 25 kDa high mobility group-1 (HMG-1) protein have generated conflicting results regarding whether HMG-1 exists as a monomer or is capable of oligomerizing to (functional) tetramers. To resolve this question, sedimentation velocity analysis yielded a s20,w value of 2.59S, which is consistent with a monomeric protein. Equilibrium sedimentation data were obtained for three HMG-1 concentrations at two rotor speeds. The six sets of data were fit to both an ideal single component and monomer-dimer equilibrium model, with essentially identical fits produced for both models, with the latter indicating a low extent (7%) of dimerization. Reaction of HMG-1 with glutaraldehyde produced a small population of oligomers consistent with a low level of dimers. This supported the monomer-dimer equilibrium model. Surprisingly, gel permeation chromatography yielded an apparent molecular mass of approx. 55 kDa for both HMG 1 and HMG-2. This finding is considered anomalous and presumably due to the high negative charge density in the C terminus of HMG-1. The sedimentation data also permit one to model HMG-1 as a hydrated prolate ellipsoid with a major axis/minor axis ratio of 2. 79. The collective evidence from the sedimentation and chemical cross-linking studies strongly supports a moderately asymmetric monomer in solution and unequivocally eliminates the possibility of a highly extended shape for HMG-1 or the existence of any extensive oligomerization. PMID- 10366724 TI - Characterisation of a family of Schistosoma japonicum proteins related to dynein light chains. AB - Dynein light chains (DLC) are components of dynein, an enzyme complex involved in various aspects of microtubule-based motility. We report here the molecular cloning and sequencing of cDNAs encoding a family of DLC-like polypeptides (SjcDLC1-5) from the human bloodfluke Schistosoma japonicum with open reading frames of 87-104 amino acids and deduced molecular masses ranging from 10.5 to 12.3 kDa. Two-dimensional Western blot analysis confirmed the presence of several S. japonicum DLC isoforms with differing pI values and molecular sizes. We also describe the molecular characterisation, genomic organisation and expression of clone SjcDLC1, and the immunological characterisation and localisation of its encoded protein. Northern blot analysis of adult worm RNA indicated SjcDLC1 is encoded by a single message of approximately 650 bp and Southern analysis suggested one SjcDLC1 gene exists in the S. japonicum genome. Immunolocalisation studies demonstrated that the SjcDLC1 protein is present in the tegument of the adult and cercarial stages of S. japonicum. SjcDLC1 and the other SjcDLC may function in the transport of specialised organelles, comprising membranous and discoid bodies, through the tegument to the schistosome-unique heptalaminate tegumental membrane at the external surface of the adult worm. As a consequence, they may provide novel targets for anti-schistosome vaccine and/or drug development. PMID- 10366725 TI - Phe-D-Leu-Phe-D-Leu-Phe derivatives as formylpeptide receptor antagonists in human neutrophils: cellular and conformational aspects. AB - We synthesized several Phe-d-Leu-Phe-d-Leu-Phe analogues in which tert butyloxycarbonyl and four different ureido substituents were included at the N terminal of the peptides, obtained as free acid and methyl-ester derivatives. Their biological action was analysed on human neutrophil responses induced by N formyl-Met-Leu-Phe (fMLF). Several in vitro assays were carried out: receptor binding, measurement of Ca2+ intracellular concentration, chemotaxis, superoxide anion production and enzyme release. A conformational investigation, using infrared absorption and circular dichroism, was also performed. Our results demonstrate that the compounds examined prefer an ordered conformation (beta turn) in amphipathic environment, and are able to antagonize the neutrophil functions evoked by fMLF. Moreover, the extent of inhibition of Ca2+ intracellular enhancement, as well as of superoxide anion production and granule enzyme release, appears related to their affinity toward the formylpeptide receptor. The free acid peptide derivatives appear to be more active antagonists than the methyl-ester ones. PMID- 10366726 TI - Rational design of a more stable yeast iso-1-cytochrome c. AB - Yeast iso-1-cytochrome c is one of the least stable mitochondrial cytochromes c. We have used a coordinated approach, combining the known functional and structural properties of cytochromes c, to engineer mutations into yeast iso-1 cytochrome c with the goal of selectively increasing the stability of the protein. The two redox forms of the native protein and six different mutant forms of yeast iso-1-cytochrome c were analyzed by differential scanning calorimetry (DSC). The relative stability, expressed as the difference in the Gibb's free energy of denaturation at a given temperature between the native and mutant forms (DeltaDeltaG(Tref)), was determined for each of the proteins. In both oxidation states, the mutant proteins C102T, T69E/C102T, T96A/C102T, and T69E/T96A/C102T were more stable than the wild-type protein, respectively. The increased stability of the mutant proteins is proposed to be due to the removal of a rare surface cysteine and the stabilization of two distorted alpha-helices. PMID- 10366727 TI - Incorporation of non-proteolytic proteins by murine alpha2-macroglobulin. AB - Human alpha2-macroglobulin is a tetrameric glycoprotein with a molecular weight of 718 kDa that is present in human plasma at high concentrations. Murine alpha2 macroglobulin is homologous to human alpha2-macroglobulin but it undergoes post translational cleavage in the subunits. Each subunit of alpha2-macroglobulin contains a thiolester which can be cleaved by small nucleophiles. In human alpha2 macroglobulin this results in a conformational change to a receptor-recognized form and a change in the electrophoretic mobility. Recent work has demonstrated that this process is reversible and during this reversal non-proteolytic proteins can become covalently trapped within the human alpha2-macroglobulin molecule. The present study further investigates this observation and examines the question whether reversal of thiolester cleavage occurs in mouse alpha2-macroglobulin. Previous studies suggest that small nucleophiles only partially convert mouse alpha2-macroglobulin to a receptor-recognized form. We demonstrate here that under appropriate conditions, mouse alpha2-macroglobulin is fully converted by NH3. We also demonstrate that despite structural and kinetic differences between human and mouse alpha2-macroglobulin, both molecules are able to incorporate non proteolytic ligands in a similar manner. This leads us to propose a general model of ligand incorporation via nucleophilic exchange in multimeric alpha macroglobulins. PMID- 10366728 TI - Bacillus thuringiensis insecticidal Cry1Aa toxin binds to a highly conserved region of aminopeptidase N in the host insect leading to its evolutionary success. AB - Bacillus thuringiensis insecticidal protein, Cry1Aa toxin, binds to a specific receptor in insect midguts and has insecticidal activity. Therefore, the structure of the receptor molecule is probably a key factor in determining the binding affinity of the toxin and insect susceptibility. The cDNA fragment (PX frg1) encoding the Cry1Aa toxin-binding region of an aminopeptidase N (APN) or an APN family protein from diamondback moth, Plutella xylostella midgut was cloned and sequenced. A comparison between the deduced amino acid sequence of PX frg1 and other insect APN sequences shows that Cry1Aa toxin binds to a highly conserved region of APN family protein. In this paper, we propose a model to explain the mechanism that causes B. thuringiensis evolutionary success and differing insect susceptibility to Cry1Aa toxin. PMID- 10366729 TI - Structure study of osteostatin PTHrP[Thr107](107-139). AB - The structure of chicken osteostatin or parathyroid hormone-related protein (PTHrP) (residues 107-139) containing an Ala/Thr substitution at the N-terminus was studied using two-dimensional proton NMR spectroscopy in an aqueous environment. Osteostatin is a separate circulating domain responsible for a range of activities related to the modulation of bone formation as well as keratinocyte proliferation. Anti-mitogenic properties of osteostatin have been detected in breast cancer cells and cytosolic calcium is used by osteostatin to signal in some neurons through a non-PTH receptor, unlike the separate circulating N terminal domain. A structural basis for the activity is presented with particular emphasis given to the conformation of the bioactive segment 107-111, forming part of a finger-like projection capable of binding to the non-PTH receptor both in the presence and absence of the remainder of the molecule which appears simply to act as a largely globular carrier. PMID- 10366730 TI - The affinity and kinetics of inhibition of cysteine proteinases by intact recombinant bovine cystatin C. AB - Recent studies have shown that the bovine cysteine proteinase inhibitor, cystatin C, is synthesized as a preprotein containing a 118-residue mature protein. However, the forms of the inhibitor isolated previously from bovine tissues had shorter N-terminal regions than expected from these results, and also lower affinity for proteinases than human cystatin C. In this work, we report the properties of recombinant, full-length bovine cystatin C having a complete N terminal region. The general characteristics of this form of the inhibitor, as reflected by the isoelectric point, the far-ultraviolet circular dichroism spectrum, the thermal stability and the changes of tryptophan fluorescence on interaction with papain, resembled those of human cystatin C. The affinity and kinetics of inhibition of papain and cathepsins B, H and L by the bovine inhibitor were also comparable with those of the human inhibitor, although certain differences were apparent. Notably, the affinity of bovine cystatin C for cathepsin H was somewhat weaker than that of human cystatin C, and bovine cystatin C bound to cathepsin L with about a four-fold higher association rate constant than the human inhibitor. This rate constant is comparable with the highest values reported previously for cystatin-cysteine proteinase reactions. The full-length, recombinant bovine cystatin C bound appreciably more tightly to proteinases than the shorter form characterized previously. Digestion of the recombinant inhibitor with neutrophil elastase resulted in forms with truncated N terminal regions and appreciably decreased affinity for papain, consistent with the forms of bovine cystatin C isolated previously having arisen by proteolytic cleavage of a mature, full-length inhibitor. PMID- 10366731 TI - Effect of ionic strength on the interfacial properties of cytochrome c. AB - The surface tension behaviour of oxidised cytochrome c (cyt c) solution was characterised at various pH and ionic strength at the air/water interface. The pendant drop method employing digital image analysis of the drop shape was applied to the measurement of the surface tension. The adsorption properties of cyt c were utilised to study the protein conformation change effected by acidification and ionic strength. At high ionic strength, the saturated steady state surface tension shows a cooperative change centred around 3.6 induced by a decrease in pH. Using spectroscopic experiments, the apparent pK of the acid induced transition of horse cyt c from the native to the molten globular state is equal to 3.5. This fact indicates that the saturated steady-state surface tension is a parameter which might be used to monitor conformation changes of cyt c. PMID- 10366732 TI - Determination of the site of disulfide linkage between heavy and light chains of silk fibroin produced by Bombyx mori. AB - The analysis of fibroin secretion-deficient 'naked-pupa' mutant silkworms has suggested that the disulfide linkage between heavy (H) and light (L) chains of fibroin, produced by the silkworm, Bombyx mori, is essential in its efficient large-scale secretion from the posterior silk gland cells. However, the site of disulfide-linkage between H- and L-chains has not been determined. In this study, cysteine residues involved in the single disulfide linkage between H- and L chains were identified as the twentieth residue from the carboxyl terminus of H chain (Cys-c20) and Cys-172 of L-chain by sequencing of genomic clones and peptide analysis. Furthermore, Cys-c4 (fourth residue from the carboxyl terminus) and Cys-c1 at the carboxyl terminus of H-chain were shown to form an intramolecular disulfide bond. PMID- 10366733 TI - Insecticidal activity of an alpha-amylase inhibitor-like protein resembling a putative precursor of alpha-amylase inhibitor in the common bean, Phaseolus vulgaris L. AB - alpha-Amylase inhibitor (alphaAI) in the common bean, Phaseolus vulgaris L., protects seeds from insect pests such as the cowpea weevil (Callosobruchus maculatus) and the azuki bean weevil (C. chinensis). Cultivars which lack alphaAI still show resistance to both bruchids. These cultivars have a glycoprotein that reacts with anti-alphaAI-1 antibodies. The glycoprotein with a molecular mass of 29 kDa (Gp29) was purified and the encoding gene was isolated. The primary structure of Gp29 is the same as alpha-amylase inhibitor-like protein (AIL) from which the encoding gene has already been isolated. AIL resembles a putative precursor of alphaAI, even though it does not form the active inhibitor. However, AIL has some inhibitory effect on the growth of C. maculatus but not C. chinensis. The presence of AIL alone is insufficient to explain the bruchid resistance of common bean cultivars lacking alpha-AI. Common bean seeds appear to contain several factors responsible for the bruchid resistance. PMID- 10366734 TI - Multi-enzyme kinetic analysis of glycolipid biosynthesis. AB - Gangliosides are acidic glycosphingolipids synthesized sequentially by a series of glycosyltransferases acting in parallel biosynthetic pathways. While most glycosyltransferases are highly specific, some, however, may catalyze equivalent steps in each pathway using different gangliosides as substrates (e.g. N acetylgalactosaminyltransferase, sialyltransferase-IV). A multi-enzyme kinetic analysis was developed on the condition that serial enzymatic reactions operate below substrate saturation. A multi-enzyme kinetic analysis enabled a simultaneous calculation of the Vmax/Km value of each enzyme derived from the equilibrium concentration of the respective substrate. Substrate concentrations [S] were determined by radioactive labelling of gangliosides in intact cells with the precursor sugars [14C]galactose and [14C]glucosamine, followed by high performance thin-layer chromatography and autoradiography of the radiolabelled glycolipids. On the basis of Michaelis-Menten kinetics, Vmax/Km values were derived from [S] by a system of linear equations. The procedure was used to analyze the development of the glycolipid composition during differentiation of rat gliomaxmurine neuroblastoma (NG108-15) cells. The Vmax/Km values calculated by multi-enzyme kinetic analysis were consistent with the kinetic data obtained with solubilized enzymes. Application of multi-enzyme kinetic analysis to published data on the correlation of enzyme activities with ganglioside levels in various cell lines and tissues indicated the validity of this method for analysis of the glycolipid biosynthesis, in particular, of its initial steps. On the basis of the kinetic analysis, it is suggested that the cell lines can be divided into two groups with respect to the substrate pools of GM3 used by sialyltransferase II and N-acetylgalactosaminyltransferase-I. The first group encompasses the majority of the neuroblastoma cell lines and the embryonic rat brain where the two enzymes share a common pool of GM3. In the second group, the two enzymes do not compete for the same pool of GM3, indicating a different subcellular localization of CMP-NeuAc:GM3 alpha2-8-sialyltransferase and UDP-N acetylgalactosaminyl:GM3 N-acetylgalactosaminyltransferase. In this study, the theory of a multi-enzyme kinetic analysis is discussed and its application to analysis of the glycolipid biosynthesis in neuroblastoma cells is demonstrated. A multi-enzyme kinetic analysis can be applied to other biosynthetic pathways and provides the advantage of analyzing kinetic data with intact cells or tissue samples. PMID- 10366735 TI - OHIO-1 beta-lactamase mutants: the Arg244Ser mutant and resistance to beta lactams and beta-lactamase inhibitors. AB - Mutations at residue 244 (Ambler numbering system) in the class A TEM beta lactamase confer resistance to inactivation by beta-lactamase inhibitors and result in diminished turnover of beta-lactam substrates. The Arg244Ser mutant of the OHIO-1 beta-lactamase, an SHV family enzyme, demonstrates variable susceptibilities to beta-lactamase inhibitors and has significantly reduced catalytic efficiency. The minimum inhibitory concentrations (MICs) for Escherichia coli DH5alpha expressing the Arg244Ser beta-lactamase were reduced when compared to the strain bearing the OHIO-1 beta-lactamase: ampicillin, 512 vs. 8192 micrograms ml-1; cephaloridine, 4 vs. 32 micrograms ml-1, respectively. The MICs for the beta-lactam beta-lactamase inhibitor combinations demonstrated resistance only to ampicillin-clavulanate, 16/8 vs. 8/4 micrograms ml-1 respectively. In contrast, there was increased susceptibility to ampicillin sulbactam, ampicillin-tazobactam, and piperacillin-tazobactam. When compared to the OHIO-1 beta-lactamase homogenous preparations of the Arg244Ser beta-lactamase enzyme demonstrated increased Km and decreased kcat values for benzylpenicillin (Km=17 vs. 50 microM, kcat=345 vs. 234 s-1) and cephaloridine (Km=97 vs. 202 microM, kcat=1023 vs. 202 s-1). Although the Ki and IC50 values were increased for each inhibitor when compared to OHIO-1 beta-lactamase, the turnover numbers (tn) required for inactivation were increased only for clavulanate. For the Arg244Ser mutant enzyme of OHIO-1, the increased Ki, decreased tn for the sulfones, and different partition ratio (kcat/kinact) support the notion that not all class A enzymes are inactivated in the same manner, and that certain class A beta-lactamase enzymes may react differently with identical substitutions in structurally conserved amino acids. The resistance phenotype of a specific mutations can vary depending on the enzyme. PMID- 10366736 TI - Nucleotide sequences of genes for ribosomal protein L41 and tRNAThr(AGU) from Coprinus cinereus. AB - The nucleotide sequences of genes for the homolog in Coprinus cinereus of the eukaryotic ribosomal protein L41 and for tRNAThr(AGU) are reported. The gene for tRNAThr(AGU) was located upstream of the gene for the L41 ribosomal protein, and these genes were adjacent to each other but in opposite orientations. The deduced amino acid sequence of ribosomal protein L41 exhibited strong homology to those of L41 proteins of several yeasts. The 56th amino acid of the deduced protein was proline, as it is in the L41 protein of a cycloheximide-sensitive strain of yeast. The putative secondary structure of the tRNA gene resembled the characteristic cloverleaf structure of tRNAs. Elements resembling an A-box and a B-box were found in the gene for tRNAThr(AGU). These boxes are known as internal promoter elements in genes for eukaryotic tRNAs. PMID- 10366737 TI - Effects of kainate-mediated excitotoxicity on the expression of rat counterparts of A170 and MSP23 stress proteins in the brain. AB - Stress proteins play important roles in the protective mechanisms under critical conditions for cell survival. We report here the expression of A170 and MSP23, oxidative stress-inducible proteins, under kainate-mediated excitotoxicity in the rat brain. A170 mRNA was significantly induced in the brain 5-8 h after i.p. kainate administration. MSP23 mRNA was observed at quite a low level in the rat brain, and the induction of MSP23 mRNA was not observed during the period 24 h after kainate administration. Immunoblot analysis demonstrated that the maximal expression level of A170 protein occurred 8 h after treatment in each part of the brain. MSP23 protein was constitutively expressed in the brain and the level of this protein was significantly decreased during the period 24 h after kainate administration. In situ hybridization and immunohistochemical studies showed that A170 was expressed predominantly in neurons, especially in pyramidal neurons of the cerebrum and cerebellar Purkinje cells, while MSP23 was expressed in oligodendrocytes. The induction of A170 was observed in the regions which are affected by excitotoxicity and this induction was observed in the earlier phase than cell death. Also, the region which shows high vulnerability to excitotoxicity such as pyramidal cell layer in the hippocampus, showed lower A170 expression than that which shows resistance to excitotoxicity, such as the dentate gyrus in the hippocampus. These results suggest that A170 may play a protective role in the brain under kainate-mediated excitotoxicity. PMID- 10366738 TI - NMDA receptor subunit gene expression in the rat brain: a quantitative analysis of endogenous mRNA levels of NR1Com, NR2A, NR2B, NR2C, NR2D and NR3A. AB - Electrophysiological recordings have shown NMDA receptors to be heterogenous structures capable of responding to selected antagonists and agonists in multiple ways. This diversity in functional response has led investigators to conclude that these channels are comprised of unique combinations of receptor subunits which determine a cell's functional NMDA-signature [H. Meguro, H. Mori, K. Araki, E. Kushiya, T. Kutsuwada, M. Yamazaki, T. Kumanishi, M. Arakawa, K. Sakimura, M. Mishina, Functional characterization of a heteromeric NMDA receptor channel expressed from cloned cDNAs, Nature (London) 357 (1992) 70-74; T. Ishii, K. Moriyoshi, H. Sugihara, K. Sakurada, H. Kadotani, M. Yokoi, C. Akazawa, R. Shigemoto, N. Mizuno, S. Nakanishi, Molecular characterization of the family of the N-methyl-d-aspartate receptor subunits, J. Biol. Chem. 268 (1993) 2836-2843; K.A. Wafford, C.J. Bain, B. Le Bourdelles, P.J. Whiting, J.A. Kemp, Preferential co-assembly of recombinant NMDA receptors composed of three different subunits, NeuroReport 4 (1993) 1347-1349; T. Priestley, P. Laughton, J. Myers, B. Le Bourdelles, J. Kerby, P.J. Whiting, Pharmacological properties of recombinant human N-methyl-d-aspartate receptors comprising NR1a/NR2A and NR1a/NR2B subunit assemblies expressed in permanently transfected mouse fiberblast cells, Mol. Pharmacol. 48 (1995) 841-848; P.H. Seeburg, N. Burnashev, G. Kohr, T. Kuner, R. Sprengel, H. Monyer, The NMDA receptor channel: molecular design of a coincidence detector, Recent Prog. Horm. Res. 50 (1995) 19-34; A.L. Buller, D.T. Monagahan, Pharmacological heterogeneity of NMDA receptors: characterization of NR1a/NR2D heteromers expressed in Xenopus oocytes, Eur. J. Pharmacol. 320 (1997) 87-94]. In situ hybridization and immunocytochemical studies have shown that there is a spatio-temporal level of expression throughout the brain for each of the receptor subunits with some regions showing a strong preference for a particular subunit. Although these studies collectively show that there are regional differences with respect to NMDA receptor subunit expression in the brain, it has not been determined at what level(s) these genes are expressed or whether each region displays a unique NMDA-subunit signature. The present study was undertaken to examine the level of gene expression for the NR1, NR2A, NR2B, NR2C, NR2D and NR3A receptor subunits in isolated regions of rat brain using the nuclease protection assay. Results show that each of the brain regions examined expresses all six NMDA receptor subunits. The level of message expression for NR1 greatly exceeded that of the other subunits combined, with values ranging from 67-88% of the total subunit gene expression. The relative proportions of the other subunits (NR2A-D and NR3A) varied widely, suggesting that NMDA receptor composition is unique to each region of the brain. PMID- 10366739 TI - Visualization of time-dependent redistribution of delta-opioid receptors in neuronal cells during prolonged agonist exposure. AB - To date, the visualization of delta-opioid receptor (DOR) internalization has been largely focused on the events of short-term agonist treatment in transfected non-neuronal cells. In this study, we followed DOR trafficking upon prolonged agonist exposure in the neuronally derived neuro2a cells, stably transfected with the fusion DOR (HA-DOR) cDNA. Internalization of surface DOR was clearly visualized in 5 min of exposure to agonist (100 nM DADLE), and the cell surface DOR remained low throughout the entire 24 h agonist exposure. Significant intracellular accumulation was visible at 20 min exposure, and increased to a maximum at 4 h, after which intracellular DOR staining gradually diminished. DOR intracellular staining was enhanced in the presence of agonist and chloroquine, a lysosomotropic agent, suggesting that internalized receptors were targeted to lysosomes and degraded upon prolonged treatment. Time-dependent colocalization of DOR with transferrin and LAMP-2 following short-term and prolonged agonist exposure further confirmed that receptor was distributed to early endosomes (sequestration) and subjected to lysosomes for degradation (down-regulation), respectively. PMID- 10366740 TI - Production and characterization of an anti-serotonin 1A receptor antibody which detects functional 5-HT1A binding sites. AB - We describe the production and characterization of a specific anti-5-HT1A receptor antibody made against a fusion protein consisting of glutathione-S transferase (GST) coupled to a 75-amino acid sequence from the middle portion of the third intracellular loop (5-HT1A-m3i, serine253-arginine327) of the rat 5 HT1A receptor protein. This region was chosen to avoid putative phosphorylation and glycosylation sites and regions of known homology with other 5-HT receptors. Western blot analysis indicated that the polyclonal anti-5-HT1A-m3i antibody accurately recognized the fusion protein expressed in bacteria and labeled a prominent 67 kDa protein band in the hippocampus, cortex, brainstem, cerebellum and kidney with a density profile corresponding to the relative abundance of the 5-HT1A receptor in these tissues. No protein was detected in liver or muscle tissue preparations, and no protein bands were labeled in any of the above tissues following preabsorption of the antibody with the 5-HT1A-m3i fusion protein. Immunohistochemistry revealed prominent labeling in limbic structures including the hippocampus, amygdala, entorhinal cortex, and septum as well as in raphe nuclei. In the hippocampus, 5-HT1A-m3i labeling revealed a characteristic laminar pattern that coincided with that seen by autoradiographic binding of the 5-HT1A agonist [3H]-8-OH-DPAT in all strata of the hippocampal formation. In the dorsal and medial raphe nuclei, anti-5-HT1A-m3i antibodies labeled the somatodendritic membranes of 5-HT neurons, consistent with its role as an autoreceptor. The detailed matching of the anti-5-HT1A-m3i antibody with [3H]-8 OH-DPAT binding suggests that the antibody recognizes a functionally active form of the 5-HT1A receptor protein capable of binding 5-HT1A agonist ligands. These anti-5-HT1A antibodies may therefore be useful tools in localizing functional 5 HT1A receptors in specific regions of the brain as well as in studying the plasticity and ontogeny of the 5-HT1A receptor at the cellular and subcellular level. PMID- 10366741 TI - Proopiomelanocortin gene expression is decreased in the infundibular nucleus of postmenopausal women. AB - Previous studies have shown that estrogen withdrawal decreases the secretion of beta-endorphin from the monkey hypothalamus. In addition, there are consistent age-associated changes in beta-endorphin neurons in the rodent. Based on these findings, we hypothesized that the activity of hypothalamic beta-endorphin neurons would be decreased in the hypothalamus of postmenopausal women. In the present study, we examined the expression of proopiomelanocortin (POMC) mRNA, the precursor mRNA for beta-endorphin, in the medial basal hypothalamus of premenopausal and postmenopausal women. Every 20th sagittal section through the hypothalamus was hybridized with a synthetic [35S]labeled, 48-base oligonucleotide probe complementary to POMC mRNA. Labeled neurons were counted and their somatic profile areas were measured with an image-combining computer microscope system. The number of POMC mRNA-containing neurons/section in the infundibular nucleus was reduced by 65% in postmenopausal women. In contrast, there was no significant difference in the number of neurons expressing POMC gene transcripts in the retrochiasmatic region. The POMC neurons in the retrochiasmatic area were also distinct morphologically from those in the infundibular nucleus. The differences between the infundibular and retrochiasmatic regions suggest that functional subgroups of POMC neurons exist in the human hypothalamus. Our findings provide evidence that the activity of hypothalamic POMC neurons is decreased in the infundibular nucleus of postmenopausal women. Both aging and gonadal steroid withdrawal may contribute to the decline in POMC gene expression in postmenopausal women. PMID- 10366742 TI - Characterization of a promoter for the human glial cell line-derived neurotrophic factor gene. AB - To address the regulation of glial cell line-derived neurotrophic factor (GDNF) gene expression, we have isolated 5' extended cDNAs, cloned the human GDNF gene, and characterized the promoter. GDNF-encoding 5' extended cDNAs containing a novel exon were isolated via reverse transcription-polymerase chain reaction (RT PCR) of mRNA from human fetal kidney and adult skeletal muscle. The GDNF gene was isolated from a human genomic library in a P1 bacteriophage vector. Analysis of the 5' flanking sequence revealed a promoter that lacks a CCAAT-box motif and is GC rich. Consensus binding sites for a variety of transcription factors have been identified in the promoter. Promoter/reporter plasmids have been constructed by fusion of the promoter and a portion of exon I to a luciferase gene. The promoter/reporter construct and a number of promoter deletions were transiently transfected into two human cell lines known to express GDNF. Multiple enhancer and silencer regions were revealed as well as a minimal promoter sufficient for basal transcription. Finally, a RT-PCR assay, specific for transcripts originating from this GDNF promoter, was developed and used to show that this promoter is active in vivo. The results suggest GDNF is regulated in a complex manner. PMID- 10366743 TI - Structure and regulation of the human NeuroD (BETA2/BHF1) gene. AB - In this study, we isolated and characterized the human NeuroD (BETA2/BHF1) gene. This gene was found to consist of two exons and one intron. The promoter regions were well-conserved compared with the mouse NeuroD gene. Two transcription start points (TSPs) were determined by the oligo-capping method. One TATA box was located at -31 bp from the lower TSP. The results of a transient transfection assay using the human neuroblastoma cell line IMR-32 and hamster insulin tumor cell line HIT-T15 suggested that there are at least three positive regulatory regions in the promoter. In these regions, four E boxes (CANNTG), named the E1 to E4 boxes, and two GC boxes were present. Cotransfection of the NeuroD expression vector into IMR-32 cells enhanced the NeuroD promoter activity by about 4-fold. A deletion and mutation analysis revealed that the E1 and E4 boxes, especially the E1 box, are associated with autoactivation and that E2 and E3 boxes are not associated with autoactivation. As mutation analysis of E3 box showed a decrease in the enhancer activity to the basal level, it showed that the E3 box is important to activate the NeuroD transcription. These results raised the possibility that the NeuroD gene expression is positively regulated through the E box sequence, not only by NeuroD itself but also by another E box binding protein. PMID- 10366744 TI - Co-expression of c-Jun and ATF-2 characterizes the surviving retinal ganglion cells which maintain axonal connections after partial optic nerve injury. AB - The expression of c-fos, c-jun, jun-b, jun-d, srf and pc4 mRNA was examined after partial optic nerve crush in the adult rat retina by in situ hybridization. Optic nerve injury led exclusively to the upregulation of c-jun, with cellular label indicative for c-jun mRNA in the retinal ganglion cell layer after two days, three days and one week post-injury. This expression pattern was in accordance with the appearance of c-Jun immunoreactivity in retinal flat mounts. Injection of an antisense but not a missense oligonucleotide against c-jun after partial crush resulted in a reduced number of connected retinal ganglion cells (RGCs) as shown by retrograde labeling. Prelabeling of RGCs with fluorogold before optic nerve section and subsequent antisense targeting against c-jun, however, led to a slightly higher number of surviving but axotomized RGCs. C-Jun antibody staining of retinal whole mounts pre- or postlabeled after crush by intracollicular administration of fluorogold showed strong c-Jun immunoreactivity in connected RGCs and also in a population of disconnected RGCs. Double labeling with an antibody directed against the transcription factor ATF-2 revealed strong co expression of c-Jun and ATF-2 in connected RGCs but not in axotomized cells. Taken together these data indicate that both RGCs in continuity and those in discontinuity with the superior colliculus respond both equally to the noxious stimulus with c-Jun expression. Moreover, the co-expression of c-Jun with high levels of ATF-2 appears to be essential for either the continuity or survival of RGCs which remain connected with their target. In disconnected RGCs, however, low levels of ATF-2 and the co-expression of c-Jun may be related to cell death. PMID- 10366745 TI - The neurotrophin receptors trkA, trkB and trkC are differentially regulated after excitotoxic lesion in rat striatum. AB - In the present work, we examined the time-dependent changes in trkA, trkB and trkC mRNA levels induced by the injection of glutamate receptor agonists into the striatum. Changes in trk mRNAs induced by quinolinate, alpha-amino-3-hydroxy-5 methyl-4-isoxazolepropionate (AMPA), kainate or 1S,3R-1-aminocyclopentane-1,3 dicarboxylic acid (ACPD) were analyzed by a ribonuclease protection assay. All high-affinity neurotrophin receptors showed differential regulation after intrastriatal injury. Up-regulation of trkA expression was observed in kainate- or ACPD-injected striata at 10 and 24 h, respectively, whereas quinolinate injection induced down-regulation between 4 and 6 h after injury. Interestingly, all the excitatory amino acid receptor agonists induced up-regulation of trkB kinase mRNA levels. This increase was maximal between 2 and 4 h after injection except in kainate injected striata, which showed the peak of expression at 10 h. In contrast, no changes in trkC mRNA expression were observed after striatal excitotoxic injury. In conclusion, our results show that trk receptor mRNA levels are differentially regulated by excitatory amino acid receptor agonists in the striatum, suggesting that changes in the levels of neurotrophin receptors might be involved either in synaptic plasticity processes or in neuronal protection in the striatal excitotoxic paradigm. PMID- 10366746 TI - Neurotensin-SPDP-poly-L-lysine conjugate: a nonviral vector for targeted gene delivery to neural cells. AB - We report herein the synthesis of a novel DNA delivery system and in vitro evidence of its ability to transfect cell lines by binding to the high-affinity neurotensin receptor and subsequent internalization of ligand-receptor complexes. The targeting vehicle consisted of neurotensin crosslinked with poly-L-lysine via N-succinimidyl-3-(2-pyridyldithio) propionate (SPDP). The SPDP-derivatives with either neurotensin or poly-L-lysine were purified by gel filtration. The conjugate resulting of the reaction of neurotensin-SPDP with HS-SPDP-poly-L lysine was purified through Biogel A 1.5. The neurotensin-SPDP-poly-L-lysine conjugate was able to bind plasmidic DNAs (pSV2cat and pGreen Lantern-1) at optimal molar ratios of 1:5 and 1:6 (DNA: conjugate), respectively. The conjugate internalized those plasmids in the cell lines (N1E-115 and HT-29) bearing the high-affinity neurotensin receptor. Expression of the plasmid products, chloramphenicol acetyltransferase and green fluorescent protein, was observed in such cell lines. Both internalization and expression of the plasmids transferred by the neurotensin-SPDP-poly-L-lysine conjugate were prevented by neurotensin (1 microM) and SR-48692 (100 nM), a specific antagonist of the high-affinity neurotensin receptor. The neurotensin-SPDP-poly-L-lysine conjugate was unable to transfect cell lines lacking the neurotensin receptor (COS-7 and L-929). In rat brain, the high-affinity neurotensin receptor is expressed by specific neurons such as those of the nigrostriatal and mesolimbic dopaminergic systems. Therefore, the neurotensin-SPDP-poly-L-lysine conjugate could be a useful tool for gene delivery to those neuronal systems. PMID- 10366747 TI - Arachidonic acid induces a long-lasting facilitation of hippocampal synaptic transmission by modulating PKC activity and nicotinic ACh receptors. AB - The present study was conducted to understand the effect of arachidonic acid on nicotinic acetylcholine (ACh) receptor-mediated synaptic plasticity. Arachidonic acid persistently (>/=1 h) potentiated currents through neuronal nicotinic ACh receptors (alpha7 and alpha4beta2) expressed in Xenopus oocytes, and the effect was blocked by the selective protein kinase C (PKC) inhibitors, such as GF109203X, PKCI, and co-expressed active PKC inhibitor peptide. This free fatty acid markedly increased nicotine-sensitive glutamate release from hippocampal slices and enhanced the rate of nicotine-sensitive miniature excitatory postsynaptic currents without affecting the amplitude in cultured hippocampal CA1 neurons under the influence of PKC. Furthermore, arachidonic acid induced a long lasting (>/=3 h) facilitation of hippocampal CA1 synaptic transmission in slices, and the effect was blocked by nicotinic ACh receptor antagonists, alpha bungarotoxin and mecamylamine. The facilitation, whereas independent of N-methyl D-aspartate (NMDA) receptors, shares a common mechanism with long-term potentiation (LTP) induced by tetanic stimulation. The results of the present study thus suggest that arachidonic acid sustains enhanced activity of nicotinic ACh receptors by interacting with a PKC pathway, thereby increasing glutamate release from presynaptic terminals, and then leading to an 'LTP-like' facilitation of hippocampal synaptic transmission. PMID- 10366748 TI - The Alzheimer-related gene presenilin 1 facilitates notch 1 in primary mammalian neurons. AB - The normal functional neurobiology of the Alzheimer's disease (AD) related gene presenilin 1 (PS1) is unknown. One clue comes from a genetic screen of Caenorhabditis elegans, which reveals that the presenilin homologue sel-12 facilitates lin-12 function [D. Levitan, I. Greenwald, Facilitation of lin-12 mediated signalling by sel-12, a Caenorhabditis elegans S182 Alzheimer's disease gene, Nature 377 (1995) 351-355]. The mammalian homologue of lin-12, Notch1, is a transmembrane receptor that plays an important role in cell fate decisions during development, including neurogenesis, but does not have a known function in fully differentiated cells. To better understand the potential role of Notch1 in mammalian postmitotic neurons and to test the hypothesis that Notch and PS 1 interact, we studied the effect of Notch1 transfection on neurite outgrowth in primary cultures of hippocampal/cortical neurons. We demonstrate that Notch1 inhibits neurite extension, and thus has a function in postmitotic mature neurons in the mammalian CNS. Furthermore, we present evidence demonstrating that there is a functional interaction between PS1 and Notch1 in mammalian neurons, analogous to the sel-12/lin-12 interaction in vulval development in C. elegans [D. Levitan, T. Doyle, D. Brousseau, M. Lee, G. Thinakaran, H. Slunt, S. Sisodia, I. Greenwald, Assessment of normal and mutant human presenilin function in Caenorhabditis elegans, Proc. Natl. Acad. Sci. U.S.A. 93 (1996) 14940-14944; D. Levitan, I. Greenwald, Effect of Sel-12 presenilin on Lin-12 localization and function in Caenorhabditis elegans, Development, 125 (1998) 3599-3606]. The inhibitory effect of Notch1 on neurite outgrowth is markedly attenuated in neurons from PS1 knockout mice, and enhanced in neurons from transgenic mice overexpressing wild type PS1, but not mutant PS1. These data suggest that PS1 facilitates Notch1 function in mammalian neurons, and support the hypothesis that a functional interaction exists between PS1 and Notch1 in postmitotic mammalian neurons. PMID- 10366749 TI - Regulation of striatal dopamine receptors by corticosterone: an in vivo and in vitro study. AB - The effects of chronic corticosterone administration and adrenalectomy on the expression of brain dopamine receptors were studied in rats. In situ hybridization and receptor binding autoradiography were carried out to determine D1, D2 and D3 receptor expression in dorsal and ventral striata. Except for down regulation of D2 mRNA in dorsal striatum after 2 week corticosterone treatment, no other significant changes were detected. In addition, the transcriptional regulation of D1 and D2 gene promoters by glucocorticoids was studied in neuroblastoma cell lines using transient transfections. While a small segment of the D2-promoter could be activated three-fold by dexamethasone, large fragments of neither D1 or D2 promoters were regulated by this treatment. Glucocorticoids do not appear to have direct overall effects on striatal dopamine receptor expression. The observed down-regulation of D2 receptor mRNA in the dorsal striatum in vivo is likely secondary to increased striatal dopamine release induced by corticosterone. PMID- 10366750 TI - Multiple splice patterns of cyclic AMP response element-binding protein mRNA in the central nervous system of the rat. AB - The alternative splicing pattern of cyclic AMP response element-binding protein (CREB) in the central nervous system (CNS) of the rat has been investigated by an exon-flanking polymerase chain reaction (PCR) strategy. A series of RT-PCR studies with primer pairs flanking all possible alternative splicing sites (corresponding to a genomic region with at least one full exon and two flanking introns) has revealed multiple splice patterns in nine regions of the rat CNS. These include some novel transcripts that lack the phosphorylation site and a segment of the leucine zipper region which is crucial for dimerization and DNA binding. Some isoforms previously reported as testis-specific were also detected in the rat CNS. The findings from this study, which include differential splicing patterns among CNS regions, suggest a complex expression and functional regulation of CREB in the CNS. PMID- 10366751 TI - Induction of the PAC1-R (PACAP-type I receptor) gene by p53 and Zac. AB - Pituitary adenylate cyclase-activating polypeptides and PAC1-R are expressed during early embryogenesis and PACAP's neurotrophic action supports a role in neuronal development. In the adult brain PACAP functions as a neuroprotective factor that attenuates the neuronal damage resulting from various insults. The tumor suppressor gene p53 and the new zinc finger protein Zac regulate apoptosis and cell cycle arrest through unrelated pathways and both genes are up-regulated under cerebral ischemia. We report here that p53 and Zac induce expression of the PAC1-R gene. By this mechanism p53 and Zac could fine-tune the balance between death promoting and protective signals and may thus fulfil a dual role in ischemia. PMID- 10366752 TI - Positive- and negative-regulation of Ca2+ entry through ACh receptor channels by phosphorylation. AB - During development of mammalian neuromuscular junctions, the expression of ACh receptors (AChRs) switches from an embryonic form to an adult form, which provides a higher Ca2+ permeable channel. The lack of this switch can prevent normal development of neuromuscular junctions. In non-mammalian species, however, the switch appears not to occur. The results of the present study show that Ca2+ entry through non-mammalian AChR channels was controlled by differential phosphorylation rather than a subunit switch. This finding provides support for the hypothesis that localized Ca2+ influx through AChR channels has a critical role in the formation and maintenance of neuromuscular junctions. PMID- 10366753 TI - Low dose of kyotorphin (tyrosine-arginine) induces nociceptive responses through a substance P release from nociceptor endings. AB - The intraplantar injection of kyotorphin (Kyo) elicited nociceptive flexor responses in mice in a dose-dependent manner between 0.1 and 100 fmol. These actions were completely blocked by substance P (NK1) receptor antagonists, such as CP-96345 and CP-99994, but not by their inactive derivatives, CP-96344 or CP 100263, nor by MEN-10376, an NK2 antagonist. Kyo-responses were also abolished by the local pretreatment with capsaicin to deplete substance P from nociceptor endings, and in tachykinin 1 gene K/O mice. These findings suggest that Kyo indirectly stimulates nociceptor endings through a local substance P release. PMID- 10366754 TI - Detection of mitochondria-derived reactive oxygen species production by the chemilumigenic probes lucigenin and luminol. AB - Both lucigenin and luminol have widely been used as chemilumigenic probes for detecting reactive oxygen species (ROS) production by various cellular systems. Our laboratory has previously demonstrated that lucigenin localizes to the mitochondria of rat alveolar macrophages and that lucigenin-derived chemiluminescence (CL) appears to reflects superoxide O2(-.) production by mitochondria in the unstimulated macrophages. In this study, we further examined the ability of lucigenin- and luminol-derived CL to assess O2(-.) and H2O2 formation, respectively, by isolated intact mitochondria. Mitochondria were isolated from monocytes/macrophages differentiated from monoblastic ML-1 cells. Incubation of the substrate-supported mitochondria with lucigenin at non-redox cycling concentration produced lucigenin-derived CL. Luminol-derived CL was also elicited with substrate-supplemented mitochondria in the presence of horseradish peroxidase (HRP). The lucigenin-derived CL was diminished extensively by the membrane permeable superoxide dismutase (SOD) mimetics, 2,2,6, 6 tetramethylpiperidine-N-oxyl and Mn(III) tetrakis(1-methyl-4-pyridyl)porphyrin, but not by Cu,Zn-SOD. On the other hand, luminol-derived CL was not observed in the absence of HRP and was significantly inhibited by catalase. A spectrum of agents known to specifically affect mitochondrial respiration exhibited corresponding effects on both lucigenin- and luminol-derived CL. Taken together, our results demonstrate that with isolated mitochondria lucigenin-derived CL monitors intramitochondrial O2(-.) production by the mitochondrial electron transport chain, whereas the luminol-derived CL detects H2O2 released from the mitochondria. As such, use of both probes provides a comprehensive and clear assessment of ROS production by mitochondria. PMID- 10366755 TI - 1-Methyladenine production from ATP by starfish ovarian follicle cells. AB - 1-Methyladenine (1-MeAde), the oocyte maturation-inducing substance in starfish, is produced by ovarian follicle cells upon stimulation with a gonad-stimulating substance (GSS) released from the radial nerves. We have shown previously that GSS causes a reduction in the intracellular levels of ATP coincident with 1-MeAde production. The present study examined whether the adenine molecule of 1-MeAde is directly derived from ATP. When isolated follicle cells from the starfish Asterina pectinifera were preloaded with [U-14C]adenine or [U-14C]adenosine, there was an increase in the intracellular levels of radiolabeled adenine nucleotides, particularly ATP. Following further incubation with GSS, the intracellular levels of radiolabeled ATP decreased, concomitant with a marked increase in the levels of [14C]1-MeAde in the medium. The amount of ATP consumed under the influence of GSS was similar to the amount of 1-MeAde produced. However, there was no change in the levels of ADP and AMP regardless of the presence or absence of GSS. These findings strongly suggest that 1-MeAde is synthesized from ATP as a substrate in follicle cells under the influence of GSS. Furthermore, using [methyl-3H]methionine, the methyl group of 1-MeAde was found to be derived from methionine. Thus GSS appears to stimulate the synthesis of 1 MeAde from ATP via the methylation process in starfish ovarian follicle cells. PMID- 10366756 TI - Binding of pepsinogen to the 78 kDa gastrin binding protein. AB - An endogenous ligand of the 78 kDa gastrin-binding protein (GBP) has been purified from detergent extracts of porcine gastric mucosal membranes by ion exchange chromatography and preparative gel electrophoresis. The ligand bound to the GBP with high affinity (mean IC50 value of 0.31+/-0.09 microgram/ml, or 8 nM), as assessed by inhibition of cross-linking of iodinated gastrin2,17 to the GBP. Both the N- and C-terminal halves of the GBP, which had been expressed individually as glutathione-S-transferase fusion proteins in Escherichia coli, and purified on glutathione-agarose beads, bound the ligand. Two peptides derived from the ligand were purified by reversed-phase high-performance liquid chromatography (HPLC), and characterised by mass spectrometry and Edman sequencing. The peptides were 97% and 100% identical, respectively, to amino acids 119-157 and 199-219 of porcine pepsinogen A. Commercial samples of pepsinogen also bound to the GBP, with a mean IC50 value of 3.9+/-1. 2 micrograms/ml (100 nM). We conclude that the ligand is closely related, but not identical, to pepsinogen A. PMID- 10366757 TI - New biodegradable hydrogels based on a photocrosslinkable modified polyaspartamide: synthesis and characterization. AB - alpha,beta-Poly(N-2-hydroxyethyl)-DL-aspartamide (PHEA), a synthetic water soluble biocompatible polymer, was derivatized with glycidyl methacrylate (GMA), in order to introduce in its structure chemical residues having double bonds and ester groups. The obtained copolymer (PHG) contained 29 mol% of GMA residues. PHG aqueous solutions at various concentrations ranging from 30 to 70 mg/ml were exposed to a source of UV rays at lambda 254 nm in the presence or in the absence of N,N'-methylenebisacrylamide (BIS); the formation of compact gel phases was observed beginning from 50 mg/ml. The obtained networks were characterized by FT IR spectrophotometry and swelling measurements which evidenced the high affinity of PHG hydrogels towards aqueous media at different pH values. In vitro chemical or enzymatic hydrolysis studies suggested that the prepared samples undergo a partial degradation both at pH 1 and pH 10 and after incubation with enzymes such as esterase, pepsin and alpha-chymotrypsin. Finally, the effect of irradiation time on the yield and the properties of these hydrogels was investigated and the sol fractions coming from irradiated samples, properly purified, were characterized by FT-IR and 1H-NMR analyses. PMID- 10366758 TI - Purification and characterization of an anti-(A+B) specific lectin from the mushroom Hygrophorus hypothejus. AB - A lectin (HHL) was isolated from the fruiting body of the mushroom Hygrophorus hypothejus by a combination of affinity chromatography on stromas of group B erythrocytes embedded in polyacrylamide gel, and DEAE-trisacryl and gel filtration chromatography. Its molecular mass, as determined by gel filtration, is estimated to be 68000 kDa and its structure is tetrameric with four identical subunits assembled with non-covalent bonds. HHL agglutinates specifically A and B blood group erythrocytes and in hemagglutination inhibition assays, exhibits sugar-binding specificity toward lactose, the anomeric alpha form being more effective than the beta form. PMID- 10366759 TI - Generation of active oxygen species from advanced glycation end-products (AGEs) during ultraviolet light A (UVA) irradiation and a possible mechanism for cell damaging. AB - Advanced glycation end-products (AGEs) have been reported to be accumulated in dermal skin. However, the role of AGEs in the photoaging of human skin remains unknown, and for this reason, we have examined the interaction between AGEs and ultraviolet A light (UVA) from both the chemical and biological aspects. Previously, we reported that exposing human dermal fibroblasts to UVA in the presence of AGEs that were prepared with bovine serum albumin (BSA) decreased the cell viability due to superoxide anion radical s (.O2(-)) and hydroxyl radicals (.OH) generated by AGEs under UVA irradiation, and active oxygen species are detected with ESR spin-trapping. To identify the active oxygen species in detail and to clarify the cell damaging mechanism, we performed several experiments and the following results were obtained. (1) In ESR spin-trapping, by addition of dimethyl sulfoxide and superoxide dismutase, ESR signals due to .O2(-) -derived DMPO-OOH and .OH-derived DMPO-OH adducts, respectively, were detectable. (2) UVA irradiated AGEs elevated the lipid peroxide levels in both fibroblasts and liposomes. But the peroxidation in liposomes was inhibited by addition of deferoxamine. (3) Survival of fibroblasts exposed to UVA in the presence of AGEs was elevated by addition of deferoxamine. And finally, (4) survival of fibroblasts was found to be regulated by the level of H2O2. On the basis of these results, we propose a possible mechanism in which AGEs under UVA irradiation generate active oxygen species involving .O2(-), H2O2, and .OH, and the .OH species plays a harmful role in promoting cell damage. PMID- 10366761 TI - Structural requirements for the binding of non-steroidal anti-inflammatory drugs to the 78 kDa gastrin binding protein. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) inhibit the proliferation of colorectal carcinoma cell lines in vitro and reduce the risk of colorectal carcinoma in vivo. The good correlation observed between the potency of NSAIDs as inhibitors of colorectal carcinoma cell proliferation and as antagonists of a 78 kDa gastrin binding protein (GBP) suggested that blockade of the GBP might contribute to the anti-proliferative effects of NSAIDs [G.S. Baldwin, V.J. Murphy, Z. Yang, T. Hashimoto, J. Pharmacol. Exp. Ther. 286 (1998) 1110-1114]. The most potent NSAID investigated was sulindac sulphide, which had an IC50 value of 40 microM. In order to investigate the structural requirements for binding to the GBP, 26 analogues of sulindac sulphide and sulindac sulphoxide were tested for their ability to inhibit the binding of iodinated gastrin to the GBP. Six of the analogues inhibited gastrin binding by more than 50% at a concentration of 1 mM. The IC50 values estimated by computer fitting of titration data were in the range of 280-940 microM. Comparison of the analogue structures suggests that a substituent with a carboxyl group is preferred in the R2 position. In addition the location of the NSAID binding site within the GBP structure was investigated. NSAIDs bound to both the N- and C-terminal halves of the GBP, and the affinities determined were similar to the values previously reported for the full-length GBP. The results reported herein represent the first step in the rational design of more potent GBP antagonists, some of which may be useful for the treatment of colorectal carcinoma. PMID- 10366760 TI - Differential behaviour of lipid based and polycation based gene transfer systems in transfecting primary human fibroblasts: a potential role of polylysine in nuclear transport. AB - DNA delivery systems for gene therapy applications have to be able to trigger the uptake of plasmid DNA into the nucleus. We have tested two types of non-viral vector systems, lipofection (cationic lipid-based, using Lipofectamine) and polyfection (cationic polymer-based, using glycerol enhanced transferrinfection), for their ability to transfect confluent, contact inhibited primary human fibroblasts. While both systems worked well with growing fibroblasts, polyfection was superior with confluent cells. A slight reduction in cell associated plasmid DNA was observed with resting cells, but it was similar for both types of complexes. Lipofectamine showed a prevalence for transfecting cycling cells as judged by costaining transfected cells with cell cycle markers. No such bias was observed when glycerol enhanced transferrinfection was used. Microinjection of plasmid DNA/polylysine complexes into the cytoplasm of fibroblasts resulted in a higher percentage of expressing cells than injection of plasmid DNA, offering an explanation for the higher transfection levels obtained with transferrinfection in non-growing cells. PMID- 10366762 TI - Isolation and characterization of a highly sulfated heparan sulfate from ascidian test cells. AB - Several sulfated polysaccharides have been isolated from the test cells of the ascidian Styela plicata. The preponderant polysaccharide is a highly sulfated heparan sulfate with the following disaccharide composition: (1) UA(2SO4)-1-->4 GlcN(SO4)(6SO4), 53%; (2) UA(2SO4)-1-->4-GlcN(SO4), 22%; (3) UA-1-->4 GlcNAc(6SO4), 14% and (4) UA-1-->4-GlcN(SO4), 11%. Two others unidentified sulfated polysaccharides and a glycogen polymer are also present in the ascidian eggs. Histochemistry with the cationic dye 1,9-dimethyl-methylene blue and biochemical analysis of the 35S-sulfate incorporation into the eggs reveal that the sulfated glycans are present exclusively in the test cells. Possibly these sulfated polysaccharides are involved in important functions of these cells, such as to confer an external and hydrophilic layer which protect the eggs and the larvae of ascidians. PMID- 10366763 TI - Development of recombinant, immobilised beta-1,4-mannosyltransferase for use as an efficient tool in the chemoenzymatic synthesis of N-linked oligosaccharides. AB - The preparation of the conserved core structure of asparagine-linked oligosaccharides found in eukaryotic glycoproteins is an important step towards the synthesis of homogeneous neoglycoproteins. So far, however, the convenient generation of the Manbeta4GlcNAcbeta4GlcNAc (Gn2M) core trisaccharide has proved to be a major obstacle because of the inherent difficulties associated with the synthesis of beta-mannosides. Here we report the overproduction in Escherichia coli of full-length and transmembrane-deleted yeast beta-1, 4 mannosyltransferases as novel N-terminal fusions bearing a decahistidinyl sequence and the minimal human Myc epitope. The recombinant enzymes were highly active and were amenable to immobilisation by nickel(II) chelation and to immunodetection with an anti-Myc monoclonal antibody. The immobilised, transmembrane-deleted enzyme exhibited an apparent Km of 14 microM for the synthetic acceptor substrate analogue, phytanyl-pyrophosphoryl-alpha-N,N' diacetylchitobioside (PPGn2), under saturating donor conditions. This figure is comparable to those previously reported for native and recombinant yeast beta-1, 4-mannosyltransferases with, respectively, the natural dolichyl-linked acceptor and PPGn2. The validity of the reaction product was confirmed by chromatographic and spectroscopic analysis. PMID- 10366764 TI - Single micro electrode dielectrophoretic tweezers for manipulation of suspended cells and particles. AB - Cells or particles in aqueous suspension close to a single capacitively coupled micro electrode (CCME) driven with high frequency electric fields experience dielectrophoretic forces. The effects near the CCME can be used for trapping and manipulation of single cells using externally metallised glass pipettes and might be used to develop a microscope based on force or capacitance measurements in conductive media. PMID- 10366765 TI - Sialic acid-specific lectin from Tritrichomonas foetus. AB - A novel sialic acid-specific lectin (TFL) was isolated from Tritrichomonas foetus culture supernatant and purified by erythrocyte adsorption followed by fetuin agarose affinity chromatography. According to gel filtration TFL is a protein of 728 kDa, different from the two sialidases of 853 and 254 kDa, secreted by T. foetus into the medium. The lectin is formed by multimeric complexes of 66 kDa subunit according to SDS-PAGE under reducing conditions. TFL is glycosylated with 4.2% of carbohydrates, half of which is represented by glucose. The lectin reacts equally with N-acetyl and N-glycolyl neuraminic acid, free, in alpha2,3- or alpha2,6-linkage. TFL has 7-fold weaker affinity to alpha2,8-linked N acetylneuraminic acid (Neu5Ac) in colominic acid. Horse erythrocytes containing 4 O-acetyl Neu5Ac are agglutinated equally as compared to the human cells. TFL affinity to 9-O-acetyl Neu5Ac is 4-fold weaker as documented by hemagglutination inhibition with de-O-acetylated bovine submaxillary mucin, and ovine submaxillary mucin. A panel of mono- and oligosaccharides other than Neu5Ac do not inhibit TFL activity at 200 mM. The lectin does not require bivalent cations for activity, shows optimal reactivity at neutral pH and is stable at 4 degrees C. Anti-TFL antibodies identify membrane positivity on T. foetus, suggesting that the lectin functions in adhesion of the parasites. These findings, together with good stability and immunogenicity, make TFL a prospective candidate for further studies, especially in searching for efficient diagnostics and prevention of bovine trichomoniasis. PMID- 10366766 TI - Urocanic acid isomers are good hydroxyl radical scavengers: a comparative study with structural analogues and with uric acid. AB - UV-exposure of the epidermis leads to the isomerisation of trans-UCA into cis-UCA as well as to the generation of hydroxyl radicals. This study shows by means of the deoxyribose degradation test that UCA isomers are more powerful hydroxyl radical scavengers than the other 4-(5-)substituted imidazole derivatives, such as histidine, though less powerful than uric acid. UCA, present in relatively high concentrations in the epidermis, may well be a major natural hydroxyl radical scavenger. PMID- 10366767 TI - Role of glutathione on acrolein-induced cytotoxicity and mutagenicity in Escherichia coli. AB - The reduced form of glutathione (GSH) is a well-known antioxidant, while there have been few reports indicating the contribution of glutathione to protection of Escherichia coli cells from the lethal effect of oxidative damage. Here, we report that depletion of glutathione causes hypersensitivity of E. coli to acrolein, the structurally simplest alpha, beta-unsaturated aldehyde derived from lipid peroxide-degradation, and that GSH can chemically react with acrolein in vitro thus reducing its toxicity. We further demonstrated that acrolein inactivates glutathione oxidoreductase followed by depletion of glutathione, probably as a part of the toxic effect of acrolein. These results suggested that GSH contributes to cellular defense against the toxic effects. PMID- 10366768 TI - Formation and persistence of DNA adducts during and after a long-term administration of 2-nitrofluorene. AB - 2-Nitrofluorene (NF) is an environmental pollutant. Our previous studies have shown that NF is a carcinogen, primarily targeting the liver, kidney and forestomach in rats. NF-induced DNA adducts were also shown higher levels in the tumor-targeting tissues compared to non-tumor targeting organs. The present study was aimed to observe the kinetics of DNA adduct formation and persistence during the process of NF-induced tumor formation. NF was supplemented in diet at three dose levels and was fed to rats continuously for up to 11 months. DNA adduct formation in the liver, kidney, spleen and stomach of rats after different period (10 days and 11 months) of NF administration was analyzed with 32P-HPLC techniques. DNA adduct persistence in the liver was also assessed after the withdrawal of NF administration. Four major NF-DNA adducts (adducts A, B, C and D) were found in the liver and kidney. DNA adduct D showed high level in the forestomach mucosa after 10 days of NF feeding while adducts A and C were undetectable. DNA adduct C and D co-migrated with C3-(deoxyguanosin-N2-yl)-2 acetylaminofluorene (dG-N2-AAF) and N-(deoxyguanosin-8-yl)-2-aminofluorene (dG-C8 AF), respectively, by 32P-HPLC co-chromatography. DNA adducts A and B constituted the major part (>80%) of NF-DNA adducts after a long period (11 months) of NF feeding. The four NF-DNA adducts showed different recovery from different enrichment procedures, i.e., nuclease P1 or butanol treatment. Three out of the four NF-DNA adducts were still detectable in the rat liver after 11 months on the basal diet. In conclusion, four major DNA adducts are induced by NF oral administration. Among those, one is identified as dG-N2-AAF and another one as dG C8-AF. The four NF-DNA adducts showed different kinetics of formation and persistence, which may play different roles in NF-induced tumor formation. PMID- 10366769 TI - Deficient nucleotide excision repair increases base-pair substitutions but decreases TGGC frameshifts induced by methylglyoxal in Escherichia coli. AB - To investigate the mutation spectrum of a well-known mutagen, methylglyoxal, and the influence of nucleotide excision repair (NER) on methylglyoxal-induced mutations, we treated wild-type and NER-deficient (uvrA or uvrC) Escherichia coli strains with methylglyoxal, and analyzed mutations in the chromosomal lacI gene. In the three strains, the cell death and the mutation frequency increased according to the dose of methylglyoxal added to the culture medium. The frequencies of methylglyoxal-induced base-pair substitutions were higher in the NER-deficient strains than in the wild-type strain, in the presence and absence of mucAB gene. Paradoxically, the frequency of methylglyoxal-induced TGGC frameshifts was higher in the wild-type strain than in the NER-deficient strains. When the methylglyoxal-induced mutation spectra in the presence and absence of mucAB gene are compared, the ratios of base-pair substitutions to frameshifts were increased by the effects of mucAB gene. In the three strains, more than 75% of the base-pair substitutions occurred at G:C sites, independent of the mucAB gene. When the mucAB gene was present, G:C-->T:A transversions were predominant, followed by G:C-->A:T transitions. When the mucAB gene was absent, the predominant mutations differed in the three strains: in the wild-type and uvrC strains, G:C-->A:T transitions were predominant, followed by G:C-->T:A transversions, while in the uvrA strains, G:C-->T:A transversions were predominant, followed by G:C-->A:T transitions. These results suggest that NER may be involved in both the repair and the fixation of methylglyoxal-induced mutations. PMID- 10366770 TI - The micronucleus assay in exfoliated buccal cells: application to occupational exposure to polycyclic aromatic hydrocarbons. AB - Many polycyclic aromatic hydrocarbons (PAHs) have been identified as cancer inducing chemicals for animals and/or humans. Also, there is sufficient evidence that exposures in the occupational settings are carcinogenic or probably carcinogenic to human. Engine exhaust and used engine oils are major PAH sources in engine repair workshops and traffic. Analysis of micronucleus (MN) in exfoliated buccal cells is a sensitive method for monitoring genetic damage in human populations. In our study, we used three different occupational groups (Group 1; engine repair workers, Group 2; taxi drivers, Group 3; traffic police) and two controls (Control I for Group 1 and Control II for Group 2 and Group 3) for the exposed groups. We analysed MN frequencies in exfoliated buccal cells and compared the exposed groups (Group 1; n=34, Group 2; n=17, Group 3; n=15) and subjects not occupationally exposed to PAH (Control I; n=28, Control II; n=20). The mean (+/-S.D.) MN (%) frequencies in exfoliated buccal cells from Group 1 and Control I were 0.07+/-0.05 and 0. 05+/-0.04, respectively (p>0.05; Table 2). The mean (+/-S.D.) MN (%) frequencies in exfoliated buccal cells from Group 2, 3 and Control II were 0.12+/-0.05, 0.10+/-0.05 and 0.03+/-0.03, respectively (p<0. 0001, p<0.05; Table 2) Smokers and nonsmokers do not differ with respect to the incidence of MN in all groups. PMID- 10366771 TI - Cytogenetic analysis of peripheral blood lymphocytes of occupational workers exposed to low levels of ionising radiation. AB - The incidence of chromosomal aberrations was analysed in peripheral blood lymphocytes of occupationally exposed people having cumulative doses of 500 mSv. The exposed individuals showed higher frequencies of dicentrics as well as acentrics than normal controls. Absorbed radiation dose was calculated by using in vitro dose response curve established for Cobalt-60 gamma rays. In the control constituting 17 healthy individuals, two dicentrics were detected among 3700 metaphases analysed. In the exposed group 27 dicentrics and one centric ring was detected among 8400 metaphases analysed. Due to small number of dicentrics scored in each individual, the dose estimate suffers from a large statistical uncertainty. The collective dose was found to be 1.89 Gy. This is in good agreement with the corrected physical doses, assuming a mean life of 10 years for the disappearance of lymphocytes. The physical doses accumulated during the last 10 years of occupation were also in good agreement with the biological dose estimate. PMID- 10366772 TI - Genotoxicity of instant coffee: possible involvement of phenolic compounds. AB - Instant coffee exhibits direct genotoxic activity in the tester strains TA 98, 100, 102, 104 and YG 1024. In the Ames tester strain TA 100, the presence of S9 mix, S100 mix, S9 mix without cofactors led to a significant decrease of the genotoxicity observed. The decrease observed in the presence of S9 mix seems to be highly correlated with the catalase content of S9 mix. The genotoxicity of instant coffee detected in strain TA 100 was dependent on the pH, with higher genotoxic effects at pH values above neutrality. Also, dependent on the pH was the ability of some phenolic molecules present in coffee promoting the degradation of deoxyribose in the presence of Fe3+/EDTA. These results suggest that apart from other molecules present in instant coffee responsible for their genotoxicity in several short term assays, phenolic molecules could also be implicated in the genotoxicity of coffee, via reactive oxygen species arising from its auto-oxidation. PMID- 10366773 TI - Detection of influenza virus induced DNA damage by comet assay. AB - Influenza virus A2/HK/68 is known to be a biological mutagen and teratogen. Reports are available implicating influenza virus as a causative agent of chromosomal aberrations in cells in culture and also in circulating leukocytes of humans. Also, an increased incidence of abortions, prenatal mortality and congenital abnormalities during the periods of epidemics has also been reported. In view of these reports, it would be worthwhile to screen persons especially pregnant women exposed to influenza virus for possible DNA damage. The present study reports the use of Comet assay to measure influenza virus induced DNA damage. We have carried out in vitro infection experiments using human leukocytes. Our results clearly indicate that influenza virus A2/HK/68 induces DNA damage in leukocytes right from 2-h post-infection. Maximum damage was observed at 24-h post-infection. However, at 48-h post-infection, a slight decrease was observed which can be attributed to the DNA repair occurring in the cells. Thereafter, irreparable damage was noticed. Cell viability results have shown lack of cytotoxicity till 72-h post-infection. However, significant cytotoxicity was observed only at 96-h post-infection. The occurrence of DNA damage without cell death may result in chromosomal aberrations or mutations. Therefore, it is most advisable to get screened for the possible DNA damage especially persons frequently infected with influenza and pregnant women. PMID- 10366774 TI - Metabolism and selected functions of sphingolipids in the yeast Saccharomyces cerevisiae. AB - Our knowledge of sphingolipid metabolism and function in Saccharomyces cerevisiae is growing rapidly. Here we discuss the current status of sphingolipid metabolism including recent evidence suggesting that exogenous sphingoid long-chain bases must first be phosphorylated and then dephosphorylated before incorporation into ceramide. Phenotypes of strains defective in sphingolipid metabolism are discussed because they provide hints about the undiscovered functions of sphingolipids and are one of the major reasons for studying this model eukaryote. The long-chain base phosphates, dihydrosphingosine-1-phosphate and phytosphingosine-1-phosphate, have been hypothesized to play roles in heat stress resistance, perhaps acting as signaling molecules. We evaluate the data supporting this hypothesis and suggest future experiments needed to verify it. Finally, we discuss recent clues that may help to reveal how sphingolipid synthesis and total cellular sphingolipid content are regulated. PMID- 10366775 TI - Recombinant adenovirus vector mediated expression of lipoprotein (a) [Lp(a)] in rabbit plasma. AB - Lipoprotein (a) [Lp(a)] is a heterodimer of apolipoprotein (a) [apo(a)] and apolipoprotein B-100 (apoB-100) of low density lipoprotein linked by a disulfide bond. Apo(a) and apoB-100 are synthesized by the liver and covalently associate or couple to form Lp(a) extracellularly. Elevated plasma Lp(a) is an independent risk factor for vascular injury disorders such as restenosis after balloon angioplasty and accelerated graft atherosclerosis following heart transplantation. Lp(a) is not expressed in laboratory animals making studies of its pathophysiology difficult. To overcome this problem, we explored the possibility of generating Lp(a) in rabbit plasma using replication-deficient adenovirus vector mediated gene delivery. Rabbits were chosen because of their large vessels and unlike mouse or rat, rabbit apoB-100 could interact with apo(a) to generate Lp(a). The recombinant (r) adenovirus vector construct used encoded a 200 kDa apo(a) [Ad-apo(a)]. Ad-apo(a) injection into the rabbit marginal vein caused the appearance of plasma rLp(a). Injection of a r adenovirus vector expressing the bacterial LacZ gene (Ad-LacZ) or PBS (vehicle) did not result in detectable plasma rLp(a). These are the first results to demonstrate plasma expression of rLp(a) in rabbits using adenovirus vector mediated gene transfer. Therefore, this system may be suitable for investigating Lp(a)'s role in the development of vascular injury diseases in a rabbit model. PMID- 10366776 TI - Biophysical and structural characterization of 1H-NMR-detectable mobile lipid domains in NIH-3T3 fibroblasts. AB - Nature and subcellular localization of 1H-NMR-detectable mobile lipid domains (ML) were investigated by NMR, Nile red fluorescence and electron microscopy, in NIH-3T3 fibroblasts and their H-ras transformants (3T3ras) transfected with a high number of oncogene copies. Substantial ML levels (ratio of (CH2)n/CH3 peak areas R=1. 56+/-0.33) were associated in untransformed fibroblasts with both (a) intramembrane amorphous lipid vesicles, about 60 nm in diameter, distinct from caveolae; and (b) cytoplasmic, osmiophilic lipid bodies surrounded by own membrane, endowed of intramembrane particles. 2D NMR maps demonstrated that ML comprised both mono- and polyunsaturated fatty chains. Lower ML signals were detected in 3T3ras (R=0.76+/-0.37), under various conditions of cell growth. Very few (if any) lipid bodies and vesicles were detected in the cytoplasmic or membrane compartments of 3T3ras cells with R<0.4, while only intramembrane lipid vesicles were associated with moderate R values. Involvement of phosphatidylcholine hydrolysis in ML generation was demonstrated by selective inhibition of endogenous phospholipase C (PC-plc) or by exposure to bacterial PC plc. This study indicates that: (1) both cytoplasmic lipid bodies and membrane vesicles (possibly in mutual dynamic exchange) may contribute (although to a different extent) to ML signals; and (2) high levels of ras-transfection either inhibit ML formation or facilitate their extrusion from the cell. PMID- 10366777 TI - Calcium-dependent PAF-stimulated generation of reactive oxygen species in a human keratinocyte cell line. AB - During inflammation and other pathological states, the lipid mediator platelet activating factor (PAF) and reactive oxygen species (ROS) are both generated. We have been investigating the effect of exogenous PAF on ROS formation in the human keratinocyte cell line (HaCaT). ROS production, measured using luminol-enhanced chemiluminescence (CL), proved to be rapid, transient, PAF receptor-mediated, and totally dependent on an increase in intracellular Ca2+ ([Ca2+]i) and on the presence of extracellular Ca2+. Repeated administration of PAF resulted in refractoriness to the agonist in terms of both capacities to increase [Ca2+]i and generate ROS. The cells, however, continued to respond fully to other stimulants (bradykinin, epidermal growth factor, thapsigargin). The PAF-induced increases in [Ca2+]i (monitored using the fluorescent probe Fluo-3) were also rapid and transient and paralleled those of ROS generation. Relatively specific inhibitors of potential ROS-producing systems were administered in an attempt to characterize the ROS producing system(s). Inhibitors of xanthine oxidase, phospholipase A2, lipoxygenase, cyclooxygenase and NO synthase did not interfere with PAF evoked ROS. The flavoprotein inhibitor diphenyleneiodonium and the mitochondrial cytochrome oxidase inhibitor KCN, prevented generation of ROS, making NAD(P)H a candidate for the electron source of the ROS and the mitochondria a potential major site of formation. PMID- 10366778 TI - Effects of linoleic acid metabolites on electrical activity in adult rat ventricular myocytes. AB - Leukotoxin (Lx), an epoxide derivative of linoleic acid, has been suggested to be a toxic mediator of multiple organ failure in burn patients and of acute respiratory distress syndrome. Lx production was recently shown during myocardial ischemia/reperfusion. However, a recent study suggested that to be toxic Lx must be metabolized to Lx-diol. In the present study, isolated adult rat ventricular myocytes were studied with the whole-cell patch-clamp technique to determine the effects of these compounds on cardiac electrical activity. Measurements of action potentials showed that neither linoleic acid nor Lx (100 microM) caused any significant changes in action potential properties. However, Lx-diol in the range of 10-100 microM produced a dose dependent increase in duration and a decrease in overshoot of the action potential. Subsequent voltage clamp experiments isolating Na current (INa) and transient outward K current (Ito) revealed that Lx-diol inhibited INa and Ito by about 80% at 100 microM, while linoleic acid and Lx had no effect on these currents at the same concentration. While Lx-diol produced the same inhibition of INa and Ito at 100 microM, its effects were more potent on Ito with significant inhibition at 10 microM. Lx-diol also hastened the activation kinetics of Ito but not INa. The action of Lx-diol was rapid (reaching steady state in 3-5 min) and was reversible in 5-10 min following washout. Thus, Lx-diol could favor arrhythmias or cardiac arrest in intact heart and may be responsible for the cardiac problems seen in systemic inflammatory response syndrome. These results further support the suggestion that Lx is not toxic in the heart but rather must be metabolized to Lx-diol to produce toxic effects on cardiac muscle. PMID- 10366779 TI - Incorporation of esterified soybean isoflavones with antioxidant activity into low density lipoprotein. AB - We have recently reported that dietary intake of soybean isoflavone phytoestrogens resulted in increased oxidation resistance of isolated low density lipoprotein (LDL). In order to explore the underlying mechanisms we designed two types of in vitro experiments. First, we prepared several different isoflavone fatty acid esters to increase their lipid solubility and studied their incorporation into LDL. Second, the oxidation resistance of the isoflavone containing LDLs was investigated with Esterbauer's 'conjugated diene' method using Cu2+ as prooxidant. Unesterified daidzein and genistein as well as genistein stearic acid esters were incorporated into LDL to a relatively small extent (0.33 molecules per LDL particle, or less) and they did not significantly influence oxidation resistance. The oleic acid esters of isoflavones were incorporated more effectively, reaching a level of 2.19 molecules per LDL particle or more, and the 4',7-O-dioleates of daidzein and genistein exhibited prolongations of lag times by 46% (P<0.05) and 202% (P<0.01), respectively. A smaller but significant increase in lag time (20.5%, P<0.01) was caused by daidzein 7-mono-oleate. In summary, esterification of soybean isoflavones daidzein and genistein with fatty acids at different hydroxyl groups provided lipophilicity needed for incorporation into LDL. Some isoflavone oleic acid esters increased oxidation resistance of LDL following their incorporation. PMID- 10366780 TI - A porcine homolog of the major secretory protein of human epididymis, HE1, specifically binds cholesterol. AB - A porcine homolog of the major secretory protein of human epididymis, HE1, was for the first time purified from the porcine cauda epididymal fluid. The HE1 homolog was secreted into the epididymal fluid as a 19-kDa glycoprotein, whose sugar moiety was gradually processed to form a 16-kDa protein during transit through the epididymis. The HE1 homolog mRNA was detected only in the caput and corpus epididymis among the porcine tissues examined. The purified HE1 homolog specifically bound cholesterol with high affinity (Kd=2. 3 microM). The binding stoichiometry was determined to be 0.94 mol/mol, suggesting that 1 mol of cholesterol binds to 1 mol of the protein. It was also found that the HE1 homolog is a major cholesterol-binding protein in the porcine epididymal fluid. The possibility that the HE1 homolog is involved in the regulation of the lipid composition of the sperm membranes during the maturation in epididymis is discussed. PMID- 10366781 TI - Novel membrane target proteins for lipoxygenase-derived mono(S)hydroxy fatty acids. AB - Hydroxyeicosatetraenoic acids (HETEs) and hydroxyoctadecadienoic acids (HODEs) are major bioactive lipids formed via the lipoxygenase oxygenation of arachidonic and linoleic acid, respectively. These metabolites appear to be involved in various cellular actions including cell proliferation, migration and regulation of enzyme activities such as phospholipases and kinases. In view of the diversity of biological effects of these hydroxy fatty acids, it seems likely that multiple mechanisms are involved. Previous reports showed that 15(S)-HETE inhibited the 5 lipoxygenase in rat basophilic leukemia (RBL-1) cell homogenates and established the presence of specific cellular HETE binding sites in these and other cells. The present study used 15(S)-HETE biotin hydrazide and 15(S)-HETE biotin pentyl amide as probes to identify membrane target proteins present in RBL-1 cells that specifically interact with HETEs and HODEs. Two membrane-associated proteins, with apparent molecular weights of 43 and 58 kDa, were identified that specifically interact with these probes and competition experiments indicated that 13(S)-HODE and 15(S)-HETE were the most effective competitors for the hydrazide probe, followed in decreasing effectiveness by 5(S)-HETE, arachidonic acid, 15(R)-HETE, stearic acid and 12(S)-HHT, a cyclooxygenase product. The two proteins were isolated and microsequencing analysis established their identities as actin and the alpha-subunit of mitochondrial ATP synthase, respectively. In vitro binding studies confirmed that purified actin is a potential 15-HETE binding protein. Subcellular cytosolic fractions exhibited fewer protein-probe complexes than membrane fractions. The association of HETEs and HODEs with these cytoskeletal and mitochondrial proteins, respectively, represents a new development in the potential actions of these hydroxy fatty acids. PMID- 10366783 TI - Natural killer cell frequency and function in Yorkshire pigs selectively bred for high or low antibody and cell-mediated immune response. AB - Porcine NK cells are small to medium-size lymphocytes having a variety of functions that may include the regulation of immune response. Thus frequency and function of NK cells were examined in generations 6 and 8 (G6, G8) of pigs selectively bred for high (H) and low (L) antibody and cell-mediated immune response (CMIR). Using the monoclonal antibody 5C6 as a pan NK phenotype marker and target cell binding and lysis as assays of function, it was found that H and L immune response pigs had high and low NK cell frequency, respectively, at G6 and G8. Function of NK cells differed by line, with the control (unselected) pigs being higher (with respect to target cell binding) or similar (with respect to lytic activity) to H which were higher than the L line pigs. Frequency of NK cells after primary immunization was in significant negative correlation with antibody after secondary immunization with hen egg white lysozyme (HEWL). Therefore, simultaneous selection for antibody and CMIR altered the frequency and function of NK cells in pigs with C > or = H > L. PMID- 10366782 TI - Inhibition by tectorigenin and tectoridin of prostaglandin E2 production and cyclooxygenase-2 induction in rat peritoneal macrophages. AB - Tectorigenin and tectoridin, isolated from the rhizomes of Korean Belamcanda chinensis (Iridaceae) which are used as Chinese traditional medicine for the treatment of inflammation, suppressed prostaglandin E2 production by rat peritoneal macrophages stimulated by the protein kinase C activator, 12-O tetradecanoylphorbol 13-acetate (TPA), or the endomembrane Ca2+-ATPase inhibitor, thapsigargin. Tectorigenin inhibited prostaglandin E2 production more potently than tectoridin. Neither compound inhibited the release of radioactivity from [3H]arachidonic acid-labeled macrophages stimulated by TPA or thapsigargin. In addition, activities of isolated cyclooxygenase (COX)-1 and COX-2 were not inhibited by the two compounds. Western blot analysis revealed that the induction of COX-2 by TPA or thapsigargin was inhibited by the two compounds in parallel with the inhibition of prostaglandin E2 production. These findings suggest that one of the mechanisms of the anti-inflammatory activities of the rhizomes of Belamcanda chinensis is the inhibition of prostaglandin E2 production by tectorigenin and tectoridin due to the inhibition of the induction of COX-2 in the inflammatory cells. PMID- 10366784 TI - Lack of natural killer cell precursors in fetal liver of Ikaros knockout mutant mice. AB - The role of Ikaros in early stages of natural killer (NK) cell differentiation was investigated using an in vitro system that promotes proliferation and differentiation of NK cell precursors into mature NK1.1+ cells. Day 14.5 fetal liver cells from mice, either homozygous for Ikaros Null or dominant negative (DN) mutations, had severe 55- to 79-fold reductions in functional NK cell precursors. Although there was no statistically significant difference between values for +/+ and +/- Null mice, the mean precursor frequency for DN mutant (+/ ) mice was significantly above that for DN -/- mice and below that for DN +/+ mice. The NK activity values for cells generated from the NK cell precursors followed the same respective relationships found for NK cell precursor frequencies. These data suggest that the deficiency of mature NK cells in Ikaros mutant mice is related to lack of functional precursors. PMID- 10366785 TI - Impaired T lymphocyte function and differential cytokine response pattern in members from cancer families. AB - In an attempt to understand the basis of lowered natural killer (NK) and T cell functions in unaffected members from cancer families, we investigated cytotoxic T lymphocyte function (CD3-directed lysis) and the ability of the lymphocytes to respond to cytokines such as IL-2, IFN-alpha and IL-12. We observed lower CD3 mediated cytotoxic activity in these individuals supported by significantly lower numbers of circulating CD3+ lymphocytes. The cytokine treatment studies revealed impaired response to IFN-alpha and IL-12 in unaffected members and breast cancer patients. The observations presented herein not only reinforce our earlier finding that lower NK and T lymphocyte function may be a feature of cancer families, but also suggest that such impaired responses may be one of the factors contributing to lower cytotoxic potential of the circulating lymphocytes. PMID- 10366786 TI - No evidence of IFN-gamma increase in the serum of HIV-positive and healthy subjects after subcutaneous injection of a non-fermented viscum album L. extract. AB - Iscador, an aqueous extract of Viscum album L., has been used for more than 80 years as an anticancer drug. Due to its immunomodulatory potential, since the onset of the AIDS epidemic it has also been applied in the treatment of HIV positive and AIDS patients in the form of the preparation V. album QuFrF (VaQuFrF; Labor Hiscia, Arlesheim, Switzerland). In in vitro investigations, incubation of peripheral blood mononuclear cells with V. album L. extracts resulted in stimulation of lymphocyte activity with increased gene expression and release of various cytokines and also of interferon gamma (IFN-gamma). In the latent phase, HIV positives exhibit only slightly elevated IFN-gamma concentrations in serum in comparison with HIV negatives, but in the acute phase of AIDS, there is an increase in levels of IFN-gamma. As the assay of cytokine levels in serum is a simple method of measuring immune system reactions, the aim of this trial was to determine whether increases in serum IFN-gamma levels in HIV positives and HIV negatives can be detected using this method after repeated injections of VaQuFrF. Five healthy subjects and 13 HIV-positive patients were investigated. IFN-gamma concentrations in serum were assayed using an ELISA test kit (ELISA test; ENDOGEN, Cambridge, Mass., USA). No drug-related elevation of serum IFN-gamma was observed at any time point during the trial. It can thus be concluded that this method is not suitable for direct investigation of the immunomodulatory effects of VaQuFrF in vivo. PMID- 10366787 TI - Natural killer cell levels in older adult mice are gender-dependent: thyroxin is a gender-independent natural killer cell stimulant. AB - The present study aimed to quantitatively analyze the effects of thyroxin on natural killer (NK) cells in the bone marrow and spleen of older adult (6 months) mice of both sexes. Five intraperitoneal injections of thyroxin were administered over 10 days in an attempt to stimulate the virtually negligible levels of NK cells in these older mice. Immunoperoxidase labeling of an NK cell surface marker, together with a tetrachrome hematologic counterstain, permitted morphological microscopic identification of mature NK cells, distinct from all other hemopoietic and immune (lymphoid) cells in the spleen and bone marrow. The results revealed that in spite of similar total cell content in both the spleen and bone marrow in both sexes at this age, there were significant, gender-based differences in NK cells and various other hemopoietic and immune cells. NK cells in the 6-month-old female mice are significantly more abundant than those of males at this age. Granulocytes in females were also significantly more numerous in both the spleen and bone marrow than in males. By contrast, monocytes and nucleated erythroid cells were significantly more numerous in males of this age. In both sexes, at this postbreeding age, the immunostimulant, thyroxin, had the common effect of significantly elevating the levels of NK cells. Our results underline the need for considering the two basic parameters of age and gender, as potentially confounding variables, prior to therapeutic administration of thyroxin, and possibly a host of hormones, cytokines and other regulators of hemopoietic and immune phenomena. PMID- 10366788 TI - Molecular basis of new disorders of iron metabolism in man -editorial-. PMID- 10366789 TI - Retroviral gene transfer into human hematopoietic cells: an in vitro kinetic study. AB - BACKGROUND AND OBJECTIVE: Successful gene therapy applications require optimized strategies to increase gene transfer efficiency into hematopietic progenitor cells (HPCs) with long-term repopulating ability. One of the issues that needs to be clarified is how hematopoietic cells proliferate, differentiate and express the transgene after each cycle of transduction. We investigated the kinetics of cell expansion, CD34 antigen expression and transduction efficiency of human hematopoietic cells in culture conditions commonly used in retroviral gene transfer protocols. DESIGN AND METHODS: Purified CD34+ cells from cord blood (n=5) or leukapheresis products (n=9) and a retroviral vector encoding an enhanced version of the green fluorescent protein (EGFP) were used. Target cells were exposed daily to vector-containing supernatants and a combination of interleukin 3 (IL-3), interleukin 6 (IL-6), stem cell factor (SCF) and Flt3 ligand (FL). Cell samples were harvested from the cultures and analyzed at 24 hour intervals for seven consecutive days. RESULTS: We found that CD34+ cells proliferated and differentiated under our culture conditions. The number of genetically modified cells increased after each cycle of transduction. Median numbers of cells positive for both CD34 and EGFP increased steadily over the culture period, but after day four most of the EGFP+ cells had a low CD34 expression. INTERPRETATION AND CONCLUSIONS: Culturing and transducing CD34+ cells for longer periods of time under these conditions might be detrimental for ex vivo gene transfer applications since the transduced cells are likely to have a decreased potential for long-term engraftment and repopulation in vivo. PMID- 10366790 TI - Recurrent mutations in the iron regulatory element of L-ferritin in hereditary hyperferritinemia-cataract syndrome. AB - BACKGROUND AND OBJECTIVE: Hereditary hyperferritinemia-cataract syndrome (HHCS) is an autosomal dominant disorder characterized by bilateral cataracts and increased serum and tissue L-ferritin, in the absence of iron overload. The deregulation of ferritin production is caused by heterogeneous mutations in the iron regulatory element (IRE) of L-ferritin that interfere with the binding of iron regulatory proteins. DESIGN AND METHODS: We have identified several patients from three unrelated Italian families with HHCS. Iron parameters were assessed by standard methods. The IRE element of L-ferritin was amplified by PCR using appropriate primers and directly sequenced. RESULTS: Ferritin levels ranged from 918 microg/L to 2490 microg/L in the patients studied. In one family bilateral cataracts were diagnosed early in life, whereas in the others cataracts were diagnosed around 40-50 years. The female proband of family 3 presented with a severe iron deficiency anemia, which was unrecognized because of the increased ferritin values. Sequencing of the IRE element of L-ferritin in the probands of the three families identified three different nucleotide substitutions (+32 GAE A, +40 AAE G and +39 CT) in the IRE of L-ferritin. These mutations have already been reported in unrelated subjects of different ethnic origins. INTERPRETATION AND CONCLUSIONS: Our findings are consistent with recurrent mutations associated with HHCS and underline the importance of this syndrome in the differential diagnosis of unexplained hyperferritinemia. In addition, the findings highlight the role played by transferrin saturation in the diagnosis of iron deficiency in these patients. PMID- 10366791 TI - Effect of glycosylation of recombinant human granulocytic colony-stimulating factor on expansion cultures of umbilical cord blood CD34+ cells. AB - BACKGROUND AND OBJECTIVE: Granulocytic colony-stimulating factor (G-CSF) is a cytokine widely used for several purposes such as stem cell mobilization, treatment of neutropenia or in vitro cultures. Recombinant human G-CSF (rh-G-CSF) is available in two forms: a non-glycosylated (E. Coli-derived), and a glycosylated CHO-derived rhG-CSF. As previously shown, glycosylation gives a higher degree of homology between the recombinant and the wild human G-CSF molecule. This study analyses the role of glycosylation in expansion cultures comparing the biological effects of the two forms of G-CSF. DESIGN AND METHODS: CD34+ cells from nine cord blood samples were positively selected (median purity 84%) and cultured in the presence of 50 ng/mL of stem cell factor and 1, 10 or 100 ng/mL of glycosylated rh-G-CSF (GG) or a deglycosylated form (DG). After 5 days of a static, serum-dependent culture fed on day 0, nucleated cells (NC), CD34+ cells and colony-forming units were evaluated and compared using the paired Student's t-test. RESULTS: For all concentrations tested, GG was able to generate more NC and progenitors than DG was able to (p<0.05). This effect was mainly observed in CFU-GM colonies, and in CFU-Mix, and indeed no influence was detected in terms of BFU-E expansion. The presence of GG in culture causes the generation of more mature granulocytic cells, assessed by the expression of CD11b/CD15 on CD13+ population, than the presence of DG. In order to check the role of the molecule's stability in this difference, the effect of daily supplementation was tested. Continuous presence of cytokines using either form of G-CSF (daily feeding) significantly increased the rate of expansion, but again GG produced higher generation than its DG counterpart. INTERPRETATION AND CONCLUSIONS: Our results suggest that the stability of the G-CSF molecule has a predominant effect on the higher biological activity found. A glycosylated form of G-CSF is recommended for in vitro cultures using serum-dependent conditions. PMID- 10366792 TI - An Italian national multicenter study for the definition of reference ranges for normal values of peripheral blood lymphocyte subsets in healthy adults. AB - BACKGROUND AND OBJECTIVE: Reference ranges are necessary in clinical chemistry and hematology to compare an observed value and to provide meaningful information. The aim of this multicenter study was the definition of reference ranges of the relative and absolute numbers of lymphocyte subsets by evaluating a large cohort of healthy adults and by using a standard protocol to reduce the variability in both sample preparation methodology and flow cytometer operation. Other aims of this study were the evaluation of the influence of sex, age, obesity, smoking, sport and some methodological variables on lymphocyte subsets and the comparison of differential white blood cell values obtained by flow cytometry and those obtained by hematology counters. DESIGN AND METHODS: Blood samples from 1311 healthy adults (blood donors and volunteers chosen according to the Italian law for donor selection) were analyzed to study, by flow cytometry, the immunophenotype of lymphocyte subsets and their distribution in terms of percentages and absolute values. Pre-analytical and analytical phases were performed according to the guidelines of the International Federation of Clinical Chemistry (IFCC) and the Italian Group of Cytometry (GIC). T cells were defined by the expression of CD3; T subpopulations by the coexpression of CD4 or CD8 or HLA-DR; B-lymphocytes were identified by the expression of CD19 while natural killer lymphocytes were identified by positivity of CD16 and/or CD56 without CD3. We calculated, for each laboratory and for all data collected, the frequency distribution percent values and absolute values of each lymphocyte subset. The influence of age, sex, smoking, obesity and sport was calculated by the t-test. The influence of some methodological variables was calculated by the t-test and multiple regression test. RESULTS: Fifty-three flow cytometry laboratories at different institutions in Italy participated in this study. Data was obtained from 1311 healthy adults aged from 18 to 70; 968 phenotype analyses (74%) were considered eligible for statistical analysis. Significant results were found as regards sex, smoking and some methodological variables (quantity of sample, washing procedures, brand of monoclonal antibodies and kind of instruments used). The comparison between hematology counters and cytometers showed no difference for any of the parameters considered. INTERPRETATION AND CONCLUSIONS: The large number of cases, the different kinds of laboratories and their distribution throughout the country make our sample representative of the Italian adult population. The standardization criteria of pre-analytical and analytical phases (the most important issues in evaluating reference values for an indicator) assured good reproducibility among laboratories so that the obtained reference ranges may be useful for interlaboratory comparison of results. Instruments and the brand of monoclonal antibodies may represent an inevitable cause of variability. PMID- 10366793 TI - Intensive salvage chemotherapy for primary refractory or first relapsed adult acute lymphoblastic leukemia: results of a prospective trial. AB - BACKGROUND AND OBJECTIVE: Adults with primary refractory or relapsed acute lymphoblastic leukemia (ALL) have a very poor prognosis with current salvage chemotherapies. Complete remissions (CR) can be obtained with intensive regimens in 40-60% of cases, but they are short-lived. In an effort to obtain high CR rates and prolong their duration and achieve long-term survival in a substantial number of patients, we designed an intensive combination salvage regimen (RELAL 88). In this protocol, chemotherapy was to be followed by an allogeneic or autologous stem cell transplant (SCT) within three months from CR. DESIGN AND METHODS: Forty-five patients with primary refractory (n=17) or first relapsed ALL (n=28) were treated with the RELAL-88 five-day induction regimen comprising vindesine, mitoxantrone, cyclophosphamide, intermediate-dose Ara-C, prednisolone and methotrexate. Twenty-eight patients received granulocyte colony-stimulating factor (G-CSF), 16 patients from day 6 (early G-CSF group) and 12 from day 14 of therapy (delayed G-CSF group). RESULTS: Thirty-four patients (74%) achieved CR (95% CI 60-87), two died in aplasia due to infection and nine were non responders. No pretreatment variable analyzed was predictive of the chance of obtaining CR. Recovery of neutrophils occurred at a median of 29 days from the start of chemotherapy without G-CSF and 20 days with G-CSF (p = 0.005), without differences between the early and late G-CSF groups. Non-hematologic side effects were usually well tolerated and consisted mainly of infections and mucositis. Twenty-three of 34 patients (68%) who achieved CR reached the planned SCT (nine autologous and 14 allogeneic). The median overall survival was 5.7 months, and the median disease-free survival for those achieving CR was 4.6 months. Of the variables analyzed for their influence on overall survival among the 34 patients who achieved CR, only the availability of an HLA-compatible sibling was associated with a prolonged survival (p = 0.03). INTERPRETATION AND CONCLUSIONS: The RELAL-88 intensive salvage regimen produces a very high rate of CR in poor risk adult ALL. Non-hematologic toxicities were tolerable, and most eligible patients could undergo the planned SCT. G-CSF significantly shortened the period of neutropenia by about eight days, irrespective of whether it was started early or late after chemotherapy. However, as with other currently available salvage therapies, remissions were short-lived, and more effective post-remission treatment strategies are needed. In our experience, only allogeneic SCT offered the chance of long-term survival. PMID- 10366794 TI - Hematopoietic damage prior to PBSCT and its influence on hematopoietic recovery. AB - BACKGROUND AND OBJECTIVE: Patients with malignancies receive chemotherapy to induce tumor remission which could damage hematopoiesis and adversely influence hematopoietic reconstitution after transplantation. In the present study we used a long-term culture (LTBMC) system and clonogenic assays to evaluate the marrow damage in patients selected to receive peripheral blood stem cell transplantation (PBSCT). DESIGN AND METHODS: Thirty-five patients - 20 with breast cancer (BC), 9 with non-Hodgkin's lymphoma (NHL) and 6 with Hodgkin's disease (HD) - were included. Bone marrow aspiration was performed one day prior to the initiation of the conditioning therapy. CFU-GM were cultured in methylcellulose with PHA-LCM. Delta assays of plastic adherent progenitor cells (PD) were performed according to Gordon's method. LTBMC were established for 5 weeks. RESULTS: There were fewer CFU-GM from all patient groups than from normal BM (p<0.05). In contrast, the number of immature progenitor cells (PD) was not decreased. The total number of CFU-GM produced by LTBMC patients was significantly reduced (p<0.05). The adherent layer from patients was often qualitatively different. In order to know whether the hematopoietic damage could affect hematopoietic reconstitution, we correlated culture data with time taken to reach peripheral cell counts. A negative correlation (r= - 0.71) was found between percentage of stromal layer and time taken to reach 20x10(9) platelets/L (tplat= 20x3-0.08% stromal layer). INTERPRETATION AND CONCLUSIONS: We can conclude that prior to PBSCT, hematopoietic function is impaired at both the level of committed progenitor cells and that of BM stroma. This damage could influence platelet recovery. PMID- 10366795 TI - Subsets of CD34+ hematopoietic progenitors and platelet recovery after high dose chemotherapy and peripheral blood stem cell transplantation. AB - BACKGROUND AND OBJECTIVE: Randomized clinical trials have shown that peripheral blood stem cell transplantations (PBSCT) with appropriate doses of CD34+ cells are associated with rapid, complete and sustained recovery of marrow functions. Nevertheless, in a minority af patients delayed platelet recovery may occur and it remains to be established whether analysis of transplanted CD34+ cell subsets may demonstrate correlation with this phenomenon. We studied a series of 80 consecutive transplanted patients with the aim of evaluating the effect of CD34+ stem cell numbers and, in a subgroup of 32 patients, the effect of the lineage specific subset numbers on time to platelet engraftment (i.e. time to platelet counts higher than 20x10(9)/L for two consecutive days without the need for platelet transfusions). DESIGN AND METHODS: Different clinical and paraclinical factors were examined in a multivariate analysis for effect on platelet engraftment in 80 patients. RESULTS: The number of CD34+ cells/kg infused was the most important factor predicting the time to platelet engraftment. Patients receiving more than 10x10(6) CD34+ cells/kg had prompt platelet engraftment. The majority of the patients (78%) received fewer than 10x10(3) CD34+ cells/kg and 17/62 (27%) of these patients experienced delayed platelet engraftment. In 32 patients receiving fewer than 10x10(6) CD34+ cells/kg we focused on the content of different lineage specific CD34+ subsets in the PBSC products. The most significant correlation was recognized for CD34+/CD61+ megakaryocytic cell number and platelet engraftment. An inverse correlation between the CD34+/CD38Eth subset and platelet engraftment was found, indicating that a high number of CD34+/CD38Eth in the PBSC product might increase the risk for delayed engraftment. These results were further confirmed by the observation that patients who experienced platelet engraftment after day 20 had significantly more CD34+/CD38Eth cells/kg infused than patients with fast engraftment. INTERPRETATION AND CONCLUSIONS: The number of total CD34+ cells/kg infused was the most important factor predicting time to platelet engraftment. CD34+ subset analysis in a subgroup of patients suggests that a high number of uncommitted progenitors may be associated with slower platelet recovery than transplantation with a higher fraction of more committed peripheral blood stem cells. PMID- 10366796 TI - HIV-HCV RNA loads and liver failure in coinfected patients with coagulopathy. AB - BACKGROUND AND OBJECTIVE: The aim of this study was to measure contemporaneously HCV-RNA load, HIV-RNA load and CD4+ lymphocyte count in HCV/HIV coinfected patients with coagulopathy and to examine the relationship between these parameters and the liver failure. DESIGN AND METHODS: A cross-sectional study was performed on 54 patients with severe coagulopathy: 39 HCV/HIV coinfected and 15 HCV+/HIV- comparable for age and HCV exposure time. HCV-RNA and HIV-RNA load, CD4+ lymphocyte count, biochemical and ultrasonographic parameters were evaluated at the time of entry to the study. RESULTS: Mean HCV-RNA load was significantly higher in coinfected patients (643,872 717,687 copies/mL) than in HCV+/HIV- (mean 161,573 276,896 copies/mL) (p = 0.01). The 39 HCV/HIV coinfected patients had a mean HIV-RNA load of 205,913 456,311 copies/mL (range 4,000-2,500,000) and a mean CD4+ lymphocyte count of 206.5171/microL (range 5-693). Five of the 39 (12.8%) coinfected patients had liver failure. In these five patients the mean HCV-RNA load (770,200 996,426 copies/mL) was high but not significantly different from that in the 34 HCV+/HIV+ patients (623,496 682,239 copies/mL) without liver failure (p = 1.0). Coinfected patients with liver failure had a significantly higher HIV-RNA load (mean 764, 599 978,542 copies/mL) and lower CD4+ lymphocyte count (mean 52.655. 6/microL) than those observed in coinfected patients without liver failure (p = 0.007 and p = 0.03, respectively). A significant inverse correlation was found between CD4+ lymphocyte count and HIV-RNA load (r = -0.37, p = 0.01). INTERPRETATION AND CONCLUSIONS: HCV-RNA load is significantly higher in HIV+ than in HIV- patients with coagulopathy. Liver failure was found only in the HCV/HIV coinfected patients with severe immunodepression, expressed either by low CD4+ lymphocyte count or by high HIV-RNA load. PMID- 10366797 TI - HLA-C and HLA-DQB1 compatibility in unrelated cord blood transplants. AB - BACKGROUND AND OBJECTIVE: Umbilical cord blood (UCB) cells have been definitively proved to be a source of hematopoietic stem cells with repopulating capacity when transplanted into pediatric hosts with neoplastic or non-neoplastic disease. Moreover, due to the immaturity of the UCB lymphoid compartment, these transplants are usually associated with a low incidence and severity of GvHD. This clinical observation and the immaturity of the UCB lymphoid compartment justify the acceptance of UCB units which differ from their recipient by 1 or 2 HLA antigens of the six HLA A, B and DRB1 antigens conventionally typed. Whether the number and type of HLA disparities affect clinical outcome of UCB transplants has not, however, been clearly demonstrated yet. DESIGN AND METHODS: In the present study on 14 pediatric patients with high risk leukemia transplanted with UCB from unrelated donors, evaluation of HLA compatibility was extended to HLA-C and DQB1 genes and correlated to the engraftment rate and occurrence of GvHD. Conditioning regimen and GvHD prophylaxis were identical in all cases. HLA-A and B antigens were typed by serology, whereas DNA based methods were used to define HLA-C gene groups, and HLA-DRB1 and DQB1 alleles. RESULTS: Conventional HLA-A, B and DRB1 typing demonstrated that 12 recipient/donor pairs differed at one HLA locus, while 2 pairs had 2 HLA disparities. The extended HLA-typing showed that only one out of the six pairs with a different HLA-A locus had additional mismatches at HLA-C and DQB1 loci, whereas all the remaining 8 pairs, which already differed at HLA-B and/or DRB1 loci after conventional typing, had additional HLA-C and/or DQB1 mismatches (p = 0.002). By contrast, engraftment rate and occurrence of GvHD did not significantly correlate with level of HLA mismatches even after extended HLA-typing. INTERPRETATION AND CONCLUSIONS: The present data show that additional mismatched HLA-C and/or DQB1 antigens are significantly more frequent in pairs which after conventional HLA-typing differed at HLA-B and/or DRB1 loci, than in those showing one HLA-A mismatch. This observation provides an additional criterion for selection of UCB donors with the closest HLA-match when more than one unit are available. We did not, however, observe any correlation between engraftment rate, occurrence of GvHD and degree of HLA disparities detected either by standard or extended typing. These data support the notion that certain HLA differences do not affect the clinical outcome of UCB transplants and indicate that the expensive and time consuming molecular typing of HLA-C and DQB1 loci might be avoided for UCB donor selection. PMID- 10366799 TI - Erythropoietin and platelet production. AB - BACKGROUND AND OBJECTIVE: Erythropoietin (Epo) is the primary growth factor for the red cell lineage but treatment with recombinant human Epo (rHuEpo) has been shown to increase platelet counts. In several animal species treatment with rHuEpo stimulated platelet production, but platelet counts tended to normalize after 1-2 weeks and large, chronic doses even caused thrombocytopenia. This paper aims to review the evidence about the effects of Epo on megakaryopoiesis. INFORMATION SOURCES: I examined the literature published in journals covered by Medline(R)a concerning the effects of Epo, hypoxia and iron deficiency on megakaryopoiesis and platelets. The reference list of each article was reviewed to try to identify further contributions. STATE OF THE ART: In vivo data have shown that moderate Epo stimulation, i.e. that produced by standard doses of rHuEpo, short-term hypoxia or moderate iron deficiency, causes a moderate elevation of platelet counts, whereas intense Epo stimulation, as produced by high doses of rHuEpo, prolonged hypoxia or severe iron deficiency, causes some degree of thrombocytopenia. In the latter case, there appears to be a diphasic response to Epo, the initial positive response (a stimulation of platelet production) being followed by thrombocytopenia. Contrarily to the thrombocytopenia due to increased platelet destruction induced by other growth factors, Epo-induced thrombocytopenia is the result of an inhibition of platelet production. CONCLUSION AND PERSPECTIVE: Stem-cell competition between erythroid and platelet precursors appears to be the cause of these phenomena in situations of prolonged, intense stimulation by Epo. In vitro data support the existence of a common erythrocytic and megakaryocytic precursor. It remains to be determined whether these effects of rHuEpo are a result of the dose itself or of the magnitude of the erythropoietic effect of that dose. It is not known whether a lower dose given in a patient with decreased marrow function would bring about the same biological effects as those induced by high doses of rHuEpo in the presence of a normal marrow function. Caution should be exercised before using high doses of hematopoietic growth factors. PMID- 10366798 TI - Diagnostic strategies in venous thromboembolism. AB - BACKGROUND AND OBJECTIVE: Diagnosis of acute deep vein thrombosis (DVT) and of pulmonary embolism (PE) is often difficult: symptomatic patients are usually investigated employing several diagnostic tests, which should be appropriately selected and sequenced, taking into account their sensitivity, specificity, safety and cost. The objective of this paper is to evaluate the performance of the new diagnostic tests and their combination in rational diagnostic strategies. DESIGN AND METHODS: A literature review was made using a Medline(R) database search for the period 1988-1998 on the following key words in various combinations: diagnosis, diagnostic strategy, venous thrombosis, pulmonary embolism, venous thromboembolism. Results of a new study by our group on diagnosis of DVT in hospitalized patients are also discussed. RESULTS: In patients with symptoms or signs suggestive of DVT, compression ultrasound (CUS) appears to be the diagnostic test of first choice, since it is a noninvasive test with high specificity and sensitivity for proximal DVT (about 97%). When CUS gives a negative result it is usually recommended that the test is repeated after one week, since its sensitivity for calf DVT is poor. The positive and negative predictive values (PPV and NPV) of CUS in symptomatic outpatients can be improved if adequate consideration is given to clinical diagnosis, using a standardized model (ref. #9), which allows symptomatic outpatients to be categorized as having a high, moderate or low probability of DVT. In case of agreement between clinical diagnosis and CUS results, no further testing is needed: patients with high or intermediate clinical probability and positive CUS results are treated, while in patients with low clinical probability and negative CUS results the diagnosis of DVT is excluded. In the case of discrepancy between clinical diagnosis and CUS results, D-dimer test and/or venography are requested. However in patients who develop signs or symptoms of DVT in the hospital the clinical model does not work, and diagnosis should be based on an appropriate mix of CUS, D-dimer (DD) test and venography. In patients presenting with signs or symptoms of pulmonary embolism, the ventilation/perfusion (V/P) lung scan remains a pivotal diagnostic test, and pulmonary angiography the reference standard, but both methods have limitations and in recent years other diagnostic tests such as echocardiography, helical (or spiral) computerized tomography, and magnetic resonance imaging have been introduced into clinical practice. Moreover, all four diagnostic tools mentioned for DVT diagnosis can be considered. Several diagnostic strategies have been proposed and evaluated in comparative studies but there is still debate over the most efficient test combination or sequence. INTERPRETATION AND CONCLUSIONS: Diagnostic strategies which include adequate consideration of clinical diagnosis using standardized models have the potential of being more efficient for outpatients (but not for inpatients) with symptoms or signs suggesting DVT of lower limbs. For patients with suspected PE, several diagnostic strategies have been assessed: V/P lung scan remains a pivotal diagnostic test, but its limitations have been increasingly recognized and newer non-invasive techniques are gaining credit. A consensus is still to be reached over the most appropriate combination of diagnostic tests. PMID- 10366800 TI - Optimizing peripheral blood progenitor cell autologous transplantation in multiple myeloma. AB - As in other malignancies, peripheral blood progenitor cells (PBPC) have almost completely replaced bone marrow as the source of stem cells for autologous transplantation in multiple myeloma. PBPC collection could be optimized either by reducing contamination by the malignant clone or by increasing hematopoietic quality of the graft. Currently, the most promising technique for purifying the harvest is CD34 cell selection. Several pilot studies have shown the feasibility of this method in MM. However controlled studies are necessary to assess the clinical impact of CD34+ cell selection. In the IFM 94 study, CD34+ selection was optional. There was no significant difference between 50 patients receiving a CD34+ selected graft and 133 patients receiving non-selected PBPC, as regards duration of neutropenia, duration of thrombocytopenia, response rate, EFS or survival. Hematopoietic recovery after transplantation is related to the number of CD34+ cells infused. The optimal regimen for mobilizing the requested CD34+ yield is not yet known. We have completed a randomized study comparing the combination of SCF plus G-CSF and G-CSF alone after priming with cyclophosphamide 4 g/m2. The median number of leukaphereses to reach the target yield of 5x10(6) CD34+ cells/kg was 1 in the SCF group (N=55) versus 2 in the G-CSF group (N=47) (p=0.008). The median number of CD34+ cells collected in the first leukapheresis was 11. 6x10(6) in the SCF group versus 4x10(6) in the G-CSF group (p=0.003). These results are in line with those observed in other trials testing the combination of SCF and G-CSF to improve PBPC collection. PMID- 10366801 TI - Cytogenetic and molecular characterization of T-cell acute lymphoblastic leukemia as a second tumor after anaplastic large-cell lymphoma in a boy. AB - We report a case of acute T-cell lymphoblastic leukemia which developed in a boy 8.5 years after successful treatment for anaplastic large-cell lymphoma. Cytogenetic and molecular characterizations of the second tumor were performed. The cytogenetic investigation revealed a complex pattern of karyotypic alterations, including double minutes, ring chromosomes, and a duplication of the p21-32 region of chromosome 1. The microsatellite DNA analysis excluded rearrangement or deletion of the TAL1 gene in the tumor cells; rearrangements of the MLL gene were excluded by Southern blot analysis. To the best of our knowledge, this is the first report of T-cell lymphoblastic leukemia arising after treatment of CD 30+ anaplastic large-cell lymphoma. The different T-cell receptor rearrangement evidenced in the two tumors indicates that this second malignancy most likely emerged de novo, but was plausibly related to the previous radiation and chemotherapy. PMID- 10366802 TI - The irreplaceable image: Paroxysmal nocturnal hemoglobinuria. PMID- 10366803 TI - The irreplaceable image: Plasmacytoma with cutaneous and pleural involvement. PMID- 10366804 TI - Hereditary hyperferritinemia cataract syndrome: a de novo mutation in the iron responsive element of the L-ferritin gene. PMID- 10366805 TI - Enumeration of CD34+ cells in fresh and thawed/washed placental/umbilical cord blood units and thawed unit segments. PMID- 10366806 TI - GM-IVA, a short induction course for de novo acute myeloid leukemia, suitable for the elderly. PMID- 10366807 TI - The differential diagnosis between aplastic anemia and hypocellular myelodysplasia in patients with pancytopenia. PMID- 10366808 TI - Parotid and central nervous system relapse during complete hematologic remission in acute promyelocytic leukemia. PMID- 10366809 TI - Prognostic irrelevance of CD34 expression in childhood B-precursor acute lymphoblastic leukemia. PMID- 10366810 TI - May-Grunwald-Giemsa fluorescence in situ hybridization technique applied to a plasma cell leukemia. PMID- 10366811 TI - Cytogenetic and in situ hybridization findings in 27 patients with atypical B cell chronic lymphocytic leukaemia. PMID- 10366812 TI - Massive hemolysis in Clostridium perfringens infection. PMID- 10366813 TI - An atypical (b3/a3) junction of the bcr/abl gene lacking abl exon a2 in a patient with chronic myeloid leukemia. PMID- 10366814 TI - Ubiquitination gene defect found in DiGeorge syndrome. PMID- 10366815 TI - New drug makes host cell machinery unavailable to HIV. PMID- 10366817 TI - Genes and environment: informing future public health decisions. PMID- 10366816 TI - Securing public access to genomic information: the race is on. PMID- 10366818 TI - Rapid update PMID- 10366819 TI - A role for phospholipase A2 in ARDS pathogenesis. AB - Acute respiratory distress syndrome (ARDS) is a life-threatening lung injury that is characterized by arterial hypoxemia and noncardiogenic pulmonary oedema. One feature of ARDS is an alteration of pulmonary surfactant that increases surface tension at the air-liquid interface and results in alveolar collapse and the impairment of gas exchange. Type-II secretory phospholipase A2 (sPLA2-II) plays a major role in the hydrolysis of surfactant phospholipids and its expression is inhibited by surfactant. Here, we discuss the evidence that in pathological situations, such as ARDS, in which surfactant is altered, sPLA2-II production is exacerbated, leading to further surfactant alteration and the establishment of a vicious cycle. PMID- 10366820 TI - The role of agouti-related protein in regulating body weight. AB - Defects in signaling by leptin, a hormone produced primarily by adipose tissue that informs the brain of the body's energy reserves, result in obesity in mice and humans. However, the majority of obese humans do not have abnormalities in leptin or its receptor but instead exhibit leptin resistance that could result from defects in downstream mediators of leptin action. Recently, two potential downstream mediators, agouti-related protein (Agrp) and its receptor, the melanocortin-4 receptor (Mc4r), have been identified. Agrp and Mc4r are excellent candidates for human disorders of body weight regulation and represent promising targets for pharmacological intervention in the treatment of these disorders. PMID- 10366821 TI - The von Hippel-Lindau tumour suppressor protein: new perspectives. AB - von Hippel-Lindau (VHL) disease is a hereditary cancer syndrome caused by germline mutations of the VHL tumour suppressor gene. The VHL gene product, pVHL, forms multiprotein complexes that contain elongin B, elongin C and Cul-2, and negatively regulates hypoxia-inducible mRNAs. pVHL is suspected to play a role in ubiquitination given the similarity of elongin C and Cul-2 with Skp1 and Cdc53, respectively. pVHL can also interact with fibronectin and is required for the assembly of a fibronectin matrix. Finally, pVHL, at least indirectly, plays a role in the ability of cells to exit the cell cycle. Thus, pVHL is a tumour suppressor protein that regulates angiogenesis, extracellular matrix formation and the cell cycle. PMID- 10366822 TI - Recent advances in the genetics of allergy and asthma. AB - Asthma is a common condition that results from the interaction of an unknown number of genes with environmental factors. About 10% of children have asthma, usually as part of a syndrome of atopy, which is characterized by the presence of allergy, asthma, seasonal rhinitis and eczema, and tends to occur in familial clusters. The incidence of asthma is lower in adults (5%) and a significant proportion is seen without an atopic background. The prevalence of asthma has increased substantially over the past decades, particularly in the western world. Allergy and asthma are not inherited as single-gene disorders and do not show a simple pattern of inheritance. Environmental and genetic factors interact in a complex fashion to produce disease susceptibility and expression. Here, we describe the recent advances in the understanding of the inherited susceptibility to asthma and atopy and discuss their potential implications. PMID- 10366823 TI - Malaria genome project task force: a post-genomic agenda for functional analysis. PMID- 10366824 TI - The epidemiology and host-parasite relationships of Schistosoma japonicum in definitive hosts. PMID- 10366826 TI - Stretching scientific intelligence on the Web PMID- 10366825 TI - Uncovering the biology and epidemiology of Neospora caninum. PMID- 10366827 TI - Deep within the filarial genome: progress of the filarial genome project. AB - Four years ago, a WHO/United Nations Development Programme/World Bank-sponsored genome project to study the filarial lymphatic nematode parasite Brugia malayi was initiated. The project took as its aims gene discovery for drug target and vaccine candidate identification, genome mapping, dissemination of genomic data to the world community and training of endemic country partners in genomic research. In this article, the principal investigators in the laboratories behind the project describe the background to the project, the data now emerging and goals for the future. Open access to filarial genome data is emphasized. PMID- 10366828 TI - The Leishmania genome comes of Age. AB - The Leishmania Genome Network (LGN) was born in Rio de Janeiro, Brazil in 1994. In the short period that has elapsed since then, the LGN has focused solely on the acquisition of the resources, and hence data, that have enabled a rational approach to genomic sequencing of the reference strain, Leishmania major Friedlin. This has now been achieved. In this review, Alasdair Ivens and Jennie Blackwell, secretary and chairman of the LGN, respectively, re-examine the approaches that were adopted, comment on some of the interesting data that have been obtained and introduce some genome-wide approaches that will facilitate functional studies of the parasite. PMID- 10366829 TI - Exploiting the life cycle of Strongyloides ratii. AB - The nematode Strongyloides ratti has a remarkable life cycle, which has both a parasitic and a free-living phase. The free-living phase includes a choice between two developmental routes. Here, Mark Viney discusses recent advances in understanding the biology of this developmental switch and shows how the life cycle of this nematode can be used to explore the lifestyle transitions common to all parasitic nematodes, as well as to address other basic biological questions. PMID- 10366830 TI - Novel secretory pathways in Plasmodium? AB - The secretion of proteins from intraerythrocytic stages of Plasmodium falciparum into the infected host cell is still poorly understood. A recent proposal that two distinct, mutually exclusive, secretory compartments may exist within the parasite cell has received much attention. Denise Mattei, Gary Ward, Gordon Langsley and Klaus Lingelbach here critically discuss the data on which this model is based, and then they address a more general question: to what extent are unusual aspects of protein secretion in Plasmodium unique among eukaryotic cells? PMID- 10366831 TI - Transmission control and drug resistance in malaria: a crucial interaction. AB - Drug resistance is a major problem affecting progress on malaria control, while many current programmes are seeking to introduce impregnated bednets to reduce transmission and hence child mortality and morbidity. David Molyneux, Katherine Floyd, Guy Barnish and Eric Fevre propose that more consideration should be given to the interaction between transmission control and the development of drug resistance, and that vector control as a means of reducing disease transmission is involved in reducing the rate of development, and the level, of resistance. Therefore, investment in vector control can have important benefits in reducing the future expenditure on drugs (as well as other costs, such as hospitalization, management of resistant cases and severe disease, drug development and household expenditure on malaria chemotherapy). Modelling the many parameters that impact on this complex relationship will better inform policy makers. PMID- 10366832 TI - Should I stay or should I go now? A stochastic model of stage differentiation in Theileria annulata. AB - The events that initiate and determine stage differentiation of protozoan parasites are not fully understood. In this article, Brian Shiels suggests that for differentiation to the merozoite in Theileria annulata the process is predetermined by the parasite, but can be initiated and modulated by changes to the extracellular environment. Shiels proposes a mechanism operating on the basis of factors that regulate gene expression reaching a commitment threshold. Similarities across protozoan and higher eukaryotic differentiation systems lead Shiels to speculate that the T. annulata model may be of relevance to other parasites. PMID- 10366833 TI - The immunoepidemiology of nematode parasites of farmed animals: a mathematical approach. AB - The population dynamics of farmed animals are controlled by humans, and often involve high host densities, which encourage higher parasite burdens than would be usual in wild animals. As a result, the immunity to reinfection acquired by the host is an important determinant of parasite population dynamics. For example, lambs are highly susceptible to gastrointestinal nematodes as they begin to graze, but develop an immunity that accounts for the observed within-year variation in parasite load and pasture contamination. In the longer term, control measures are compromised by the development of parasite strains resistant to chemotherapy, focusing attention on the development of 'natural' measures, including the selection for resistant hosts and the development of antiparasite vaccines. Mick Roberts here considers the immunoepidemiology of parasites of farmed animals on three levels: the interaction between the parasite and the host's immune system determining the individual's level of protection; the development of acquired immunity determining the within-year parasite population dynamics; and the long-term effects of control measures on the between-year parasite population dynamics. PMID- 10366834 TI - Haptoglobin, the acute phase response and natural human immunity to trypanosomes. PMID- 10366835 TI - Reply PMID- 10366836 TI - Publish and be damned! PMID- 10366837 TI - Expanding market for urinary incontinence therapies. PMID- 10366838 TI - Lipid-based amphotericin B formulations: from animals to man. AB - Amphotericin B has been the mainstay of systemic antifungal therapy for over 30 years, despite its serious side-effects, and, although numerous alternative antifungal agents have been developed, none to date has matched the efficacy of amphotericin B. However, modern drug delivery technology has improved the safety of amphotericin B by incorporating it into lipid-based delivery systems, including liposomes. Three such formulations, based on the natural affinity of amphotericin B for lipids, are currently marketed. All increase the therapeutic index of amphotericin B, thereby allowing more aggressive treatment than is possible with the conventional product. However, they differ in structure, side effect profiles and evidence of proven efficacy as discussed in this review. PMID- 10366839 TI - Excipients as stabilizers. AB - Excipients are better known as promoters of degradation than as stabilizers of drug substances. This is not surprising. Functional groups or residues in excipients can have the propensity to interact with labile active ingredients, with attendant loss of molecular integrity or other changes in quality. Thus, the canon of work on excipients as stabilizers is not extensive. Nevertheless, possibilities exist to capitalize on our knowledge of how a drug substance degrades, and of the properties and composition of excipients, to convert unstable drugs into viable products. This article discusses such approaches to product stabilization. PMID- 10366840 TI - Methodology for pharmacokinetic/pharmacodynamic data analysis. AB - This review will highlight the role of exploratory data analysis and nonlinear regression software in pharmacokinetic-pharmacodynamic (PK/PD) data analysis. Kinetic and dynamic modelling situations typical to the pharmacokineticist in the drug industry will be addressed, and two case studies, including single-dose intravenous bolus plasma data and multiple-dose response-time data will be analysed. Approaches to assessing the suitability of model fit to the observed data will be discussed, and a summary of the key features of available commercial PK software will be given. Specific emphasis will be placed on the application of WinNonlin. PMID- 10366841 TI - Trade secrets and other R&D capital preservation techniques. AB - In today's age of ballooning R&D expenses, it is critical to preserve and make the most cost-effective use of discoveries and expertise. Business and marketing plans make positive use of such discoveries and expertise through creating and selling products based upon this intellectual property. Legal protections preserve the discoveries and expertise, by preventing competitors from profiting from the efforts of innovative companies. PMID- 10366842 TI - Monitor: progress and profiles. PMID- 10366843 TI - A natural example of protein trans-splicing. PMID- 10366845 TI - Plasmon resonance spectroscopy: probing molecular interactions within membranes. AB - Surface plasmon resonance (SPR) has become a popular method for investigating biomolecular interactions. A new variant of this technique, coupled plasmon waveguide resonance (CPWR) spectroscopy, allows the characterization of anisotropic biological membranes. Plasmon resonance can therefore be used to study the molecular events involved in a wide variety of membrane processes, including energy conversion and signal transduction. PMID- 10366844 TI - Remote contributions to subunit interactions: lessons from adult and fetal hemoglobins. PMID- 10366846 TI - The ambivalence of vitamin E in atherogenesis. AB - The early events in atherogenesis might be due to the oxidation of low- density lipoprotein. The antioxidant vitamin E, therefore, has received much attention as a potential anti-atherogenic agent. Recent mechanistic studies of the early stage of lipoprotein-lipid oxidation show that the role of vitamin E in this process is not simply that of a classical antioxidant. Unless additional compounds are present, vitamin E can have antioxidant, neutral or pro-oxidant activity. This more complex function is reflected in the results of vitamin-E-intervention studies of atherosclerosis in animals and of controlled prospective trials on the incidence of cardiovascular disease in humans, which, overall, are inconclusive. PMID- 10366847 TI - Ceramide and apoptosis. PMID- 10366848 TI - The use of diglyceride kinase for quantifying ceramide. PMID- 10366849 TI - Reply to kolesnick and hannun, and perry and hannun PMID- 10366850 TI - Ceramide second messengers and ceramide assays. PMID- 10366851 TI - A novel transactivation domain in parkin. PMID- 10366852 TI - The Ca-calmodulin-dependent protein kinase cascade. AB - The Ca2+-calmodulin-dependent protein kinase (CaM kinase) cascade includes three kinases: CaM-kinase kinase (CaMKK); and the CaM kinases CaMKI and CaMKIV, which are phosphorylated and activated by CaMKK. Members of this cascade respond to elevation of intracellular Ca2+ levels and are particularly abundant in brain and in T cells. CaMKK and CaMKIV localize both to the nucleus and to the cytoplasm, whereas CaMKI is only cytosolic. Nuclear CaMKIV regulates transcription through phosphorylation of several transcription factors, including CREB. In the cytoplasm, there is extensive cross-talk between CaMKK, CaMKIV and other signaling cascades, including those that involve the cAMP-dependent kinase (PKA), MAP kinases and protein kinase B (PKB; also known as Akt). Activation of PKB by CaMKK appears to be important in protection of neurons from programmed cell death during development. PMID- 10366853 TI - Mammalian Kruppel-like transcription factors: more than just a pretty finger. AB - The transcription factor SP1 contains three Kruppel-like zinc fingers. Recently, several related proteins, including erythroid, lung and gut-enriched Kruppel-like factors, have been identified. Together with SP1, these proteins form a sizeable family of transcription factors that share homology in their zinc-finger domains but differ elsewhere. Analysis of these differences is illuminating specific mechanisms by which transcription is regulated. PMID- 10366854 TI - Selection, history and chemistry: the three faces of the genetic code. AB - The genetic code might be a historical accident that was fixed in the last common ancestor of modern organisms. 'Adaptive', 'historical' and 'chemical' arguments, however, challenge such a 'frozen accident' model. These arguments propose that the current code is somehow optimal, reflects the expansion of a more primitive code to include more amino acids, or is a consequence of direct chemical interactions between RNA and amino acids, respectively. Such models are not mutually exclusive, however. They can be reconciled by an evolutionary model whereby stereochemical interactions shaped the initial code, which subsequently expanded through biosynthetic modification of encoded amino acids and, finally, was optimized through codon reassignment. Alternatively, all three forces might have acted in concert to assign the 20 'natural' amino acids to their present positions in the genetic code. PMID- 10366855 TI - Back to Camelot: defining the specific role of tRNA in protein synthesis. PMID- 10366856 TI - Salmonella typhimurium recognition of intestinal environments. PMID- 10366857 TI - Salmonella typhimurium recognition of intestinal environments: response PMID- 10366858 TI - Time for a fresh look at the bacterial chromosome. PMID- 10366859 TI - Transforming growth factor beta in infectious disease: always there for the host and the pathogen. AB - At the portals of pathogen entry, there are pools of the latent form of a potent cytokine, transforming growth factor beta (TGF-beta). Many infections activate these pools and stimulate further TGF-beta expression. As well as potent immunomodulation, activated TGF-beta might have important effects on pathogen entry, replication, persistence, latency and oncogenesis. PMID- 10366860 TI - Perspective: phloem transport of viruses and macromolecules - what goes in must come out. AB - Phloem transport of endogenous macromolecules and plant viruses remains poorly understood. Selective movement into and out of the phloem is tightly regulated, yet the mechanisms governing this selectivity have not been elucidated. Recent advances in identifying nonviral proteins and nucleic acids with the capacity for phloem transport will hopefully shed new light on the regulation of phloem loading and unloading. PMID- 10366861 TI - Protective mechanisms against toxic electrophiles in Escherischia coli. AB - Escherichia coli are exposed to toxic electrophiles from both endogenous and exogenous sources. To survive, E. coli have developed novel protective mechanisms that appear unique to bacteria. In the absence of one of these mechanisms, electrophiles induce rapid killing. Hence it appears that electrophile-protective mechanisms represent novel targets for antibacterial drug research. PMID- 10366862 TI - Molecular mechanisms of yeast longevity. AB - The genetic analysis of yeast longevity has illuminated the underlying molecular mechanisms of aging that invoke the importance of metabolic regulation, genetic stability and stress resistance in determination of life span. The RAS genes have emerged as important modulators of life-maintenance processes and of life span itself. PMID- 10366863 TI - Bacterial S-layers. AB - S-layers are produced by the self assembly of proteinaceous subunits on the surfaces of prokaryotes, so that planar, monomolecular-thick crystalline lattices are formed. Some archaeal and eubacterial S-layer proteins are glycosylated. These lattices typically have center-to-center spacings of less than 25 nm, which makes them attractive for biomimetic or nanotechnological applications. PMID- 10366864 TI - The involvement of leptin in humans revealed by mutations in leptin and leptin receptor genes. PMID- 10366865 TI - Two novel families of ectonucleotidases: molecular structures, catalytic properties and a search for function. PMID- 10366866 TI - G protein antagonists. AB - Heterotrimeric G proteins couple membrane-bound heptahelical receptors to their cellular effector systems (ion channels or enzymes generating a second messenger). In current pharmacotherapy, the input to G protein-regulated signalling is typically manipulated by targeting the receptor with appropriate agonists or antagonists and, to a lesser extent, by altering second messenger levels, most notably by inhibiting phosphodiesterases that hydrolyse cyclic nucleotides. When stimulated, G proteins undergo a cycle of activation and deactivation in which the alpha-subunits and the betagamma-dimers sequentially expose binding sites for their reaction partners (receptors, guanine nucleotides and effectors, as well as regulatory proteins). These domains can be blocked by inhibitors and this produces effects that cannot be achieved by receptor antagonists. Here, the structural and mechanistic information on G protein antagonists is summarized and an outline of the arguments supporting the hypothesis that G proteins per se are also potential drug targets is provided. PMID- 10366867 TI - Molecular manipulations as tools for enhancing our understanding of 5-HT neurotransmission. AB - A developing trend in exploring the sites at which drugs act is to use molecular rather than chemical agents to alter receptors, intracellular signalling mechanisms or gene expression. The 5-HT neurotransmission system is targeted by drugs useful in many behavioural disorders, including anxiety, depression, psychosis and eating disorders. It also regulates many physiological functions and provides some examples of the potential use of these new molecular approaches. This article reviews the progress made in the molecular manipulation of 5-HT receptors and discusses the potential of such tools for the treatment of diseases associated with the 5-HT transmission system. PMID- 10366868 TI - Corrigendum PMID- 10366869 TI - Pontine regulation of pelvic viscera: pharmacological target for pelvic visceral dysfunctions. AB - The pathophysiology and pharmacological targets of disorders of the bladder and colon have focused predominantly on the periphery. However, these viscera are regulated by the CNS, which, in turn, must integrate their functions with compatible behaviours. This review focuses on the role of the pontine micturition centre, Barrington's nucleus, as a key to this integration. Through its efferent network this pontine centre links parasympathetic preganglionic neurones with forebrain-projecting nuclei, providing an anatomical substrate for coregulation of pelvic visceral and forebrain activity. Disorders characterized by multiple pelvic visceral symptoms and comorbidity with psychiatric disorders (for example functional bowel disorders) might have their roots in dysfunctions of this circuit, which could provide a novel target for pharmacological treatment. PMID- 10366870 TI - Gene therapy for lung disease. AB - Gene therapy is a new field of medical research that has great potential to influence the course of treatment of human disease. The lung has been a particularly attractive target organ for gene therapy due to its accessibility and the identification of genetic deficits for a number of lung diseases. Several clinical trials have shown evidence of low levels of gene transfer and expression, but without any benefit to the patients involved. Thus, current studies are focusing on further research and technological improvements to the vectors. Gene therapy is now beginning to benefit from a shift in emphasis from clinical trials to the development of better tools and procedures to deliver gene therapy to the bedside. PMID- 10366871 TI - Wood formation in forest trees: from Arabidopsis to Zinnia/IT> PMID- 10366872 TI - Syntaxin on the brain. PMID- 10366874 TI - Signalling in gene silencing. PMID- 10366873 TI - Jumping genes and maize genomics. PMID- 10366875 TI - Stephen Hales and the cohesion theory. PMID- 10366876 TI - Regulatory mechanism of plant gene transcription by GT-elements and GT-factors. AB - GT-elements are regulatory DNA sequences ususally found in tandem repeats in the promoter region of many different plant genes. Depending on promoter structure, GT-elements can have a positive or a negative transcription function. The cognate GT-element binding factors contain one or two trihelix DNA binding motifs, which have so far been identified in plant transcription factors only. GT-factors are ubiquitously expressed; in Arabidopsis they belong to a small family of transcription factors. The functioning of plant GT-elements and GT-factors shows complex regulatory features of plant gene transcription. PMID- 10366877 TI - Defense on multiple fronts: how do plants cope with diverse enemies? AB - Plants have evolved an array of defense mechanisms to protect themselves against the wide variety of pathogens and pests with which they are confronted. Included in these defense mechanisms are inducible responses that are turned on systemically in the plant in response to attempted infection or predation. The two most studied inducible responses are systemic acquired resistance, which provides enhanced resistance to pathogen infection, and the wound response pathway, resulting in enhanced resistance to insect feeding. Recent research suggests that the two pathways are not completely independent, and the induction of one might affect the expression of the other. However, the evidence for cross talk between different induced defense response pathways is somewhat confusing, and at times contradictory. Here, we review recent advances in our understanding of how the different pathways might interact. PMID- 10366878 TI - 'Radicle' biochemistry: the biology of root-specific metabolism. AB - The roots of higher plants are a fascinating and largely unexplored biological frontier. One of their features is the ability to synthesize a remarkable diversity of secondary metabolites, and to adjust their metabolic activities in response to biotic and abiotic stress. This includes the ability to exude a complex array of micro- and macromolecules into the rhizosphere, with the potential to affect the inter-relationships between plants and beneficial or deleterious soil-borne organisms. In the past, research on root biology has been hampered by the underground growth habit of roots and by the lack of a suitable experimental system. However, recent progess in growing roots in isolation has greatly facilitated the study of root-specific metabolism and contributed to our understanding of this remarkable plant organ. PMID- 10366879 TI - Recent advances in the transformation of plants. AB - Plant transformation technology has become a versatile platform for cultivar improvement as well as for studying gene function in plants. This success represents the culmination of many years of effort in tissue culture improvement, in transformation techniques and in genetic engineering. The next challenge is to develop technology that minimizes or eliminates the tissue culture steps, and provides predictable transgene expression. PMID- 10366880 TI - Carotenoid sequestration in plants: the role of carotenoid-associated proteins. AB - In plants, carotenoid accumulation and sequestration take place within chloroplasts and chromoplasts. In the chloroplast, practically all carotenoids are associated with chlorophyll-binding proteins, whereas chromoplasts have developed a unique mechanism to sequester carotenoids within specific lipoprotein structures. Recent research into the existence of a group of homologous genes that encode carotenoid-associated proteins that aid in the generation of carotenoid-lipoprotein structures in chromoplasts, offers a new framework for elucidating the carotenoid sequestration mechanism. PMID- 10366881 TI - A guide to the Lhc genes and their relatives in Arabidopsis/IT> AB - The Lhc super-gene family encodes the light-harvesting chlorophyll a/b-binding (LHC) proteins that constitute the antenna system of the photosynthetic apparatus, and also includes some relatives whose functions are more or less unknown. The Lhc super-gene family of Arabidopsis contains >30 members and the databases contain >1000 EST clones originating from these genes. This article presents an overview of these genes and provides some tools for researchers who want to use them in their studies. PMID- 10366883 TI - Border control thwarts particles entering the brain. PMID- 10366882 TI - A primary ovarian leiomyosarcoma with micro-invasive features (stage I): is surgical excision enough? PMID- 10366884 TI - Retraction by Jeff Schell. PMID- 10366885 TI - Advances in CE/MS. PMID- 10366886 TI - Nanoparticle detection technology for chemical analysis. PMID- 10366887 TI - Analysis at home. PMID- 10366888 TI - Infrared multiphoton dissociation in an external ion reservoir. AB - In this work we present a novel scheme for performing infrared multiphoton dissociation (IRMPD) external to the mass analyzer in an external ion reservoir consisting of an rf-only multipole and a pair of electrostatic lens elements. Ions generated by electrospray ionization (ESI) are accumulated in an rf-only hexapole and dissociated by irradiation at 10.6 microns from a CW CO2 laser in the source region of the mass spectrometer. This scheme is unique from other IRMPD schemes as dissociation occurs in a spatially distinct region of the spectrometer and is independent of the mass spectrometry platform used to analyze the fragment ions. The effectiveness of the technique is demonstrated with ESI IRMPD FTICR mass spectrometry of a 20-mer phosphorothioate oligonucleotide. A comparison of the external IRMPD scheme with nozzle-skimmer dissociation and conventional in-cell IRMPD reveals a significant improvement in signal-to-noise ratio and fragment yield, particularly for larger, more highly charged fragment ions. PMID- 10366889 TI - Cellular applications of a sensitive and selective fiber-optic nitric oxide biosensor based on a dye-labeled heme domain of soluble guanylate cyclase. AB - Nitric oxide-selective sensors have been prepared with the heme domain of soluble guanylate cyclase (sGC), the only known receptor for signal transduction involving nitric oxide. Expressed in and purified from E. coli, the heme domain contains a stoichiometric amount of heme that has electronic and resonance Raman spectra almost identical to those of heterodimeric (native) sGC purified from bovine lung. The small size of the heme domain, its inability to bind oxygen, and its high affinity for nitric oxide make it well-suited for sensor applications. The heme domain has been labeled with a fluorescent reporter dye and changes in this dye's intensity are observed based on the sGC heme domain's characteristic binding of nitric oxide. The current sensors are prepared with 100-microns optical fiber but could also be prepared using submicrometer fiber tips. These sensors have fast, linear, and reversible responses to nitric oxide and are unaffected by numerous common interferents, such as oxygen, nitrite and nitrate. The sensor limit of detection is 1 microM nitric oxide. Glutathione has been shown to decrease the sensitivity of the sensor; however, the sensor response remains linear and can be calibrated on the basis of the glutathione concentration present in the biological environment of interest. The sensors have been used to measure extracellular nitric oxide production by BALB/c mouse macrophages. Minimal nitric oxide was produced by untreated cells, while high levels of nitric oxide were released from activated cells, e.g., 111 +/- 2 microM in a given cell culture. PMID- 10366890 TI - Probing proteomes using capillary isoelectric focusing-electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry. AB - Unlike the genome, the proteome is exquisitely sensitive to cellular conditions and will consist of proteins having abundances dependent upon stage in the cell cycle, cell differentiation, response to environmental conditions (nutrients, temperature, stress etc.), or disease state(s). Therefore, the study of proteomes under well-defined conditions can provide a better understanding of complex biological processes and inference of protein function. Thus, much faster, more sensitive, and precise capabilities for the characterization of cellular constituents are desired. We describe progress in the development and initial application of the powerful combination of capillary isoelectric focusing (CIEF) and Fourier transform ion cyclotron resonance (FTICR) mass spectrometry for measurements of the proteome of the model system Escherichia coli. Isotope depletion of the growth media has been used to improve mass measurement accuracy, and the comparison of CIEF-FTICR results for the analysis of cell lysates harvested from E. coli cultured in normal and isotopically depleted media are presented. The initial studies have revealed 400-1000 putative proteins in the mass range 2-100 kDa from total injections of approximately 300 ng of E. coli proteins in a single CIEF-FTICR analysis. PMID- 10366892 TI - Convergent, self-encoded bead sensor arrays in the design of an artificial nose. AB - We report a new approach to designing an artificial nose based on high-density optical arrays that directly incorporate a number of structural and operational features of the olfactory system. The arrays are comprised of thousands of microsphere (bead) sensors, each belonging to a discrete class, randomly dispersed across the face of an etched optical imaging fiber. Beads are recognized and classified after array assembly by their unique, "self-encoded" response pattern to a selected vapor pulse. The high degree of redundancy built into the array parallels that found in nature and affords new opportunities for chemical-sensor signal amplification. Since each bead is independently addressable through its own light channel, it is possible to combine responses from same-type beads randomly distributed throughout the array in a manner reminiscent of the sensory-neuron convergence observed in the mammalian olfactory system. Signal-to-noise improvements of approximately n1/2 have been achieved using this method. PMID- 10366891 TI - Direct, ultrasensitive, and selective optical detection of protein toxins using multivalent interactions. AB - Three highly sensitive, selective, and reagent-free optical signal transduction methods for detection of polyvalent proteins have been developed by directly coupling distance-dependent fluorescence self-quenching and/or resonant-energy transfer to the protein-receptor binding events. The ganglioside GM1, as the recognition unit for cholera toxin (CT), was covalently labeled with fluorophores and then incorporated into a biomimetic membrane surface. The presence of CT with five binding sites for GM1 causes dramatic change for the fluorescence of the labeled GM1. (1) In the scheme using fluorescence self-quenching as a signal transduction mechanism, the fluorescence intensity drops significantly as a result of aggregation of the fluorophore-labeled GM1 on a biomimetic surface. (2) By labeling GM1 with a fluorescence energy transfer pair, aggregation of the labeled GM1 results in a decrease in donor fluorescence and an increase in acceptor fluorescence, providing a unique signature for selective protein receptor binding. (3) In the third scheme, using the biomimetic surface as part of signal transduction and combining both fluorescence self-quenching and energy transfer mechanisms to enhance the signal transduction, a signal amplification was achieved. The detection systems can reliably detect less than 0.05 nM CT with fast response (less than 5 min). This approach can easily be adapted to any biosensor scheme that relies on multiple receptors or co-receptors. The methods can also be applied to investigate the kinetics and thermodynamics of the multivalent interactions. PMID- 10366893 TI - Hydroquinone-based derivatization reagents for the quantitation of amines using electrochemical detection. AB - Two new reagents, NDTE (2,5-dihydroxyphenylacetic acid, 2,5-bis-tetrahydropyranyl ether p-nitrophenyl ester) and HLTE (homogentisic gamma-lactone tetrahydropyranyl ether), are described for the chemical derivatization of primary and/or secondary amines to form an electrochemically active product. These reagents undergo reaction with the aforementioned analytes to form a product possessing the hydroquinone moiety, thus allowing for reversible electrochemical detection at mild oxidation potentials. The reactivity of each reagent was demonstrated by using N-ethylbenzylamine (EBzA) and the dipeptide isoleucine leucine methyl ester as model analytes. The investigation included the isolation and identification of the intermediates and final products from derivatization of EBzA. These isolated standards were subsequently characterized with respect to electrochemical properties by means of cyclic voltammetry. In LC-EC experiments, the concentration limit of detection (CLOD) of the purified EBzA product was determined to be 5 nM (100 fmol) at a detection potential of +200 mV vs Ag/AgCl ([Cl-] = 3 M). The CLOD values obtained by LC-EC after derivatization of aqueous solutions of EBzA and Ile-Leu-OMe with NDTE were 25 nM (250 fmol) and 250 nM (2.5 pmol), respectively. PMID- 10366894 TI - An electrochemiluminescence flow-through cell and its applications to sensitive immunoassay using N-(aminobutyl)-N-ethylisoluminol. AB - An electrochemiluminescence (ECL) flow-through cell with a carbon fiber electrode for ECL detection was developed, and a flow injection analysis system containing the cell was established. N-(aminobutyl)-N-ethylisoluminol (ABEI), an ECL reagent, was determined by this system. A straight-line calibration curve for ABEI (r = 0.999) was obtained from 6 fmol to 25 pmol. The detection limit of ABEI was 6 fmol (S/N = 2) and the relative standard deviation was 1.7% at 1.5 pmol (n = 10). The system was used for immunoassay of human immunogloblin G (hIgG) with ABEI-labeled anti-hIgG. A calibration curve of hIgG was obtained from 80 pg/mL to 1.3 ng/mL. The detection limit of hIgG was 80 pg/mL (S/N = 2), and the relative standard deviation was 1.8% at 0.5 ng/mL (n = 10). Sensitivity and accuracy for sera samples were found to considerably exceed those of the conventional methods, such as single-radial immunodiffusion and nepherometric immunoassay. The present system should prove useful for immunoassay. PMID- 10366895 TI - On-line technique for measuring stable oxygen and hydrogen isotopes from microliter quantities of water. AB - Detailed here is a method for extracting and analyzing oxygen and hydrogen isotopes from 10 microL-sized water samples. Based on the traditional CO2-H2O equilibration technique, the oxygen isotope exchange reaction is done exclusively in sealed 6-mm (o.d.) Pyrex tubes at 25 degrees C, with full isotope exchange completed in at least 28 h. Using the same water sample employed in the 18O equilibration, D/H extractions are done in separate sealed 6-mm (o.d.) Pyrex tubes by reaction with Zn at 450 degrees C to form H2(g). Provided that a correction factor is applied to 18O analyses, accuracy and precision for both 18O and D/H are comparable to standard techniques using much larger samples. PMID- 10366896 TI - Biopersistence and durability of nine mineral fibre types in rat lungs over 12 months. AB - The study objectives were to assess the ability of intratracheal injection methods to discriminate between nine fibre types in respect of pulmonary biopersistence, and to provide approximate estimates of relative biopersistence and durability for a study of general relationships with biological and toxicological responses. The test fibres included six samples of size-selected fibre types specially prepared for research purposes, two commercially available fibres, and amosite. A 1 mg dose of each fibre type was administered to rats by intratracheal injection. The relative biopersistence of fibres in different size categories was assessed from the changes in mean lung burden, as determined by electron microscopy, at 3 days and 1, 6 and 12 months after injection. The ability of the test materials to resist dissolution was measured in a parallel series of simple in vitro acellular experiments at two pHs and in a continuous flow dissolution test. The observed differences in the persistence of fibres of differing length recovered from rat lungs were consistent with the current hypothesis that short fibres are cleared by cellular processes and long fibres by dissolution and disintegration. Differences in persistence of long (> 20 microns) fibres were correlated with measured rates of dissolution in vitro. Differences in persistence among those fibre types also studied by others workers were consistent with their findings after inhalation and intratracheal injection. Overall, the differences in the biopersistences of the test fibres following intratracheal injection were sufficient to enable an examination of the relationship of biopersistence with other biological and toxicological responses. Biopersistence was influenced by both fibre dimensions and solubility. PMID- 10366897 TI - Influence of fibre length, dissolution and biopersistence on the production of mesothelioma in the rat peritoneal cavity. AB - A range of respirable man-made mineral fibres were tested for evidence of carcinogenicity by injection into the peritoneal cavity of male SPF Wistar rats; and differences in carcinogenicity were related to the dimensions and biopersistence of the injected fibres. The fibres tested included an amosite asbestos, a silicon carbide whisker, a special purpose glass microfibre, and a range of other man-made vitreous fibres (MMVFs) and refractory ceramic fibres (RCFs) from the TIMA fibre repository. The injected dose of each was designed as the estimated mass required to contain 10(9) fibres > 5 microns in length, as determined by optical microscopy. The numbers of long fibres (> 15 microns) contained in these doses ranged across fibres from 0.1 x 10(9) to 0.8 x 10(9) fibres; the number of long fibres thinner than 0.95 micron ranged from 0.015 x 10(9) to 0.4 x 10(9). The treatment groups contained between 18 and 24 animals. Animals were killed when they showed signs of debilitation. At autopsy, the diagnosis of mesothelioma was usually obvious macroscopically. Otherwise, histological examination of peritoneal organs was used to search for early tumour development. Judged by median survival time, four of the fibre types, in the doses administered, presented higher mesothelioma activity than amosite asbestos. The other fibres tested were less carcinogenic than the amosite. Only a ceramic material derived by extreme heating to simulate the effect of furnace or oven conditions, produced no mesotheliomas. Attempts were made, using regression models, to relate these differences to fibre dimensions and to measures of durability from separate experiments. The results pointed principally to a link with the injected numbers of fibres > 20 microns in length and with biopersistence in the rat lung of fibres longer than 5 microns. Improved quantification of the relative importance of fibre dimensions and biopersistence indices requires experimentation with a range of doses. PMID- 10366899 TI - The effects of strapped spectacles on the fit factors of three manufactured brands of full facepiece negative pressure respirators. AB - A study was conducted to determine the effects of strapped spectacles on the fit factors obtained during quantitative fit testing on three different brands of full facepiece negative pressure respirators. The three brands of respirators were evaluated with and without strapped spectacles worn by the test subjects. A total of 180 quantitative fit testing trials were conducted on ten male test subjects. For each test subject, three quantitative fit testing trials were performed with each brand of respirator with and without the spectacles. The average of the fit testing trials for each subject with each respirator was used for statistical analysis. The results demonstrated that the fit factor values were significantly lower during use of the spectacles (p < 0.05). The estimated percentage of test subjects who failed the American National Standards Institute pass/fail criteria for quantitative fit testing (1000) increased by 15-36% when spectacles were worn. PMID- 10366898 TI - Influence of characteristics of inhaled fibres on development of tumours in the rat lung. AB - The objective was to examine and quantify the influence of fibre dimensions, persistence in the lung, and dissolution and cell toxicity in vitro, on the risks of developing lung tumours in rats. Data were brought together from the studies carried out at the IOM under the Colt Fibre Research Programme, and from studies carried out in Switzerland and the USA under the programme of the Thermal Insulation Manufacturers Association. In both studies, groups of rats were exposed by inhalation to a range of airborne fibres. At the end of their lives they were examined for the presence of benign and malignant lung tumours and mesothelioma. The studies differed in a number of details, but were combined on the basis of approximate equivalence of cumulative exposure to airborne fibres. Logistic regression models were used to relate differences in carcinogenicity to fibre characteristics; dimensions, persistence in the lung after intratracheal injection, dissolution rates from bench-top flow-through experiments, measures of inflammation, and other cell responses to fibres in vitro. Despite the small number of data points, the results suggested a primary influence of the airborne concentrations of the numbers of fibres thinner than 1 micron diameter and longer than 20 microns, and of the measured dissolution rate of the fibres. While these results are based on only a small number of fibre types, the statistical model fits the data reasonably well, and enables some cautious insights into the quantitative influences of dimensions and biopersistence. Results were broadly consistent with those from intraperitoneal injection studies of the same fibres, in that the responses were dependent on both the durability of the fibres and the numbers of long thin fibres. In vitro and in vivo cell responses did not predict significantly the risk of cancer following inhalation. PMID- 10366900 TI - Immunomodulating action and structure-activity relationships of substituted phenylamides of 5-amino-3-methylisoxazole-4-carboxylic acid. AB - A series of 5-amino-3-methylisoxazole-4-carboxylic acid amides has been prepared by condensation of 5-amino-3-methylisoxazole-4-carboxylic acid with ethyl chloroformate. The resulting mixed anhydride undergoes condensation with appropriate phenylamides to form the corresponding amides 6-16. The compounds obtained were evaluated for their immunological activities in cultures of human peripheral blood mononuclear cells (PMBC). We found that the activities of the compounds in the proliferation test and in the lipopolysaccharide (LPS)-induced cytokine production in PBMC cultures were differential. The stimulatory or inhibitory effects depended strongly on the origin and location of substituents in the phenyl ring which is described in the discussion and was supported by QSAR studies. PMID- 10366901 TI - Inhibition of human immunodeficiency virus type (HIV-1) replication by some diversely functionalized spirocyclopropyl derivatives. AB - Inhibition of HIV-1 replication by differently substituted spirocyclopropyl compounds has been evaluated. Compound 21 showed a moderate activity (EC50 ranging from 2.3 to 5.8 micrograms/ml) against different HIV-1 strains (IIIB, RF, NDK, and an AZT-resistant strain) in different cell lines (MT-4 and C-8166 cells), while it was cytotoxic at 77.7 micrograms/ml, resulting in a selectivity index of 34. This compound was inactive against HIV-2 (ROD) and SIV (MAC251). From "time-of-addition" experiments compound 21, like AZT, appeared to inhibit HIV-1 at the reverse transcriptase step. PMID- 10366902 TI - Synthesis, physicochemical properties, anticonvulsant activities, and GABA-ergic and voltage-sensitive calcium channel receptor affinities of alpha-substituted N benzylamides of gamma-hydroxybutyric acid. Part 4: Search for new anticonvulsant compounds. AB - In a search for new anticonvulsant compounds, two series of N-benzylamides of alpha-(benzylamino)-gamma-hydroxybutyric acid (series A) and alpha-(2 phenylethylamino)-gamma-hydroxybutyric acid (series B), were investigated in maximal electroshock (MES), subcutaneous metrazole, and rotorod toxicity assays. The most potent anticonvulsant compounds were alpha-(benzylamino)-gamma hydroxybutyric acid N-benzylamide (3) and N-(2-chlorobenzylamide (4) with median effective (ED50) doses 63.0 mg/kg and 54.0 mg/kg, respectively. alpha-(4 Phenylpiperazinyl)-gamma-hydroxybutyric acid N-(4-methylbenzyl)amide (17) and alpha-(benzylpiperazinyl-gamma-hydroxy-butyric acid N-(4-methylbenzyl)amide (18) were also tested for their ability to potentiate [3H]-muscimol binding and to inhibit [35S]-TBPS binding (as indices of GABA-A receptor potentiation). Amide 17 exhibited activity at the GABA-A complex which may be the mechanism by which the anticonvulsant effect of this compound is mediated. The N-benzylamides of alpha (benzylamino)-gamma-hydroxybutyric acid (3-9) were also evaluated for their ability to displace [3H]-nitrendipine from voltage-sensitive calcium channel (VSCC) receptors isolated from rat cortex. PMID- 10366903 TI - Pharmacokinetic investigations with direct injection of plasma samples: possible savings using capillary electrophoresis (CE). AB - Capillary electrophoresis (CE) is often regarded as a separation technique of choice because of its high selectivity and its cost advantages compared to LC.RSD% of 0.5% have become standard for quality control assays. Using CE, sample pretreatment can often be significantly reduced, leading to notable savings of labor and reagent costs. Moreover, errors from sample pretreatment steps are avoided. A number of pharmaceuticals (e.g. acetaminophen, salicylic acid, sulfamethoxazole, theophylline, tolbutamide, and trimethoprim) have been determined in human plasma on underivatized fused silica capillaries by MEKC without sample pretreatment, the total analysis time being only 10 min. An sodium dodecyl sulfate-containing borate buffer (60 mM with 200 mM SDS) at pH 10 has been used. Between runs, proteins adsorbed to the capillary wall are removed by a rinsing regimen consisting of SDS buffer and either acetonitrile (e.g. 50% v/v) or isopropanol (e.g. 10% v/v). Other rinsing approaches are discussed (salts, enzyme containing solutions, organic solvents, sodium hydroxide, hydrofluoric acid). The separation system is tested in a concentration range between 10 ng/mL and 100 micrograms/mL, the detection limit being about 5 ng/mL. The sensitivity has been substantially improved compared to preceding work using field-amplified injection mechanisms and efficient computer algorithms that take advantage of multiwavelength detection. Correlations between the limit of quantitation (LOQ), the limit of detection (LOD) and the signal/noise ratio are discussed. A day-to day precision for relative peak areas of 1 to 2% relsdv (n > 40) has been reached in the upper concentration range. Thus, not only drug monitoring but also pharmacokinetic investigations from blood plasma have become possible without further sample pretreatment. PMID- 10366904 TI - Synthesis and biological activities of dibenzyl dipiperazine diquaternary ammonium salts. AB - Twenty new 4,4'-dibenzoyl-1,1'-dibenzyl-1,1'-(decane-1,5-diyl)-piperazi nium dihalides 5a-l and 1,1'-dibenzyl-1,1'-(decane-1,5-diyl)piperazinium dihydrochloride dihalides 6a-l were prepared and evaluated for their analgesic, sedative and anti-inflammatory activities. Structure-activity relationship studies indicated that the compounds 6c (Ar = 4-NO2C6H5) and 6k (Ar = 3-Me-C6H5) exhibited higher activities than others. Compared with the corresponding compounds 6k and 6l, the presence of benzoyl in the compound 5k and 5l exerted a contrary influence on the activities. 5h and 6h show the highest anti inflammatory activity (59%, dose 20 mg/kg and 48%, dose 1 mg/kg) in the series 5 and 6, respectively. PMID- 10366905 TI - Interaction of Viagra with the NO donors molsidomine and RE 2047 with regard to antithrombotic and blood pressure lowering activities. AB - After p.o. administration to rats in doses up to 30 mg/kg, Viagra showed no antithrombotic effect. However, it enhanced the inhibition of thrombus formation by RE 2047 from 9% to 17% (5 + 5 mg/kg) or 19% to 27% (10 + 10 mg/kg) in arterioles. This effect was even more obvious in venules where an inhibition of 9% (5 + 5 mg/kg) or 15% (10 + 10 mg/kg) was seen whereas the individual drugs had no effect. The antithrombotic activity of molsidomine was not altered. The blood pressure (b.p.) of spontaneously hypertensive rats was reduced by the combination of Viagra and RE 2047 (5 + 5 mg/kg) to 94% of normal after 2 h while the individual drugs had no effect at this dose. The coadminstration of 10 mg/kg of each drug reduced the b.p. to 87% of normal. The combination of Viagra with molsidomine decreased b.p. to 84% (5 + 5 mg/kg) or 79% (10 + 10 mg/kg), respectively. PMID- 10366906 TI - The development of an implant for the metacarpophalangeal joint of the fingers. PMID- 10366907 TI - Osseointegrated silicone implants. 18 patients with 57 MCP joints followed for 2 years. AB - 20 patients were operated on consecutively with osseointegrated MCP joint prostheses in 64 joints at our department between September 1993 and February 1995. The one-stage procedure included joint resection and cancellous bone grafting from the iliac crest before insertion of screw-shaped titanium fixtures, connected with a flexible silicone spacer. 18 patients (57 joints) were clinically and radiographically examined at median 28 (18-37) months postoperatively. Indications for surgery were joint destruction due to chronic arthritis in 17 patients (56 joints), and posttraumatic arthrosis in 1 patient (1 joint). Postoperative median range of motion was 40 (15-85) degrees, with an extension deficit of 30 (-20-70) degrees. 16 patients were satisfied, and had good pain relief and substantially improved postoperative hand function, evaluated with the standardized Sollerman hand function test. Radiographic osseointegration was obtained in 112 of 114 titanium fixtures (98%), but fracture of the silicone spacer was observed in 14 implants (25%). We conclude that osseointegration of longitudinal titanium fixtures in the bone marrow canal is possible in a one-stage procedure, but our findings show the need for a new, more durable joint spacer. PMID- 10366908 TI - Pin-tract complications in external fixation of fractures of the distal radius. AB - We analyzed retrospectively the rate and outcome of pin-tract complications in 314 unstable fractures of the distal radius, treated with the Hoffmann small frame external fixator. The overall rate of complications was 27%. The commonest complication (21%) was pin-tract infection, which was treated with oral antibiotics. There were no cases of osteitis. Complications led to premature removal of the fixator in 17 of the cases. Women over the age of 75 years had a significantly higher rate of pin loosening (17%), but not a higher rate of premature removal of the fixator due to complications. 4% of the cases had a pin site fracture, all women. The rate of pin-tract complications was high, but severe complications were rare, even in old women. PMID- 10366909 TI - Trans-styloid fixation of fractures of the distal radius. A prospective randomized comparison between 6- and 1-week postoperative immobilization in 60 fractures. AB - We performed a prospective randomized study on 60 patients with dorsally displaced extra-articular or noncomminuted intraarticular fractures of the distal radius. All 60 fractures were treated by closed reduction and Kirchner wire trans styloid fixation. 30 patients had 1 weeks' postoperative immobilization and 30 patients had 6 weeks' immobilization. All patients had a clinical and radiographic review at 6 weeks and at 1 year after the operation. Pain, range of movement and grip strength were tested clinically, and changes in dorsal tilt, frontal radial deviation, ulnar variance, and radial shortening were assessed radiographically. Rates of complications were the same in both groups. At follow up, pain was similar in both groups and range of motion and grip strength were somewhat better after early mobilization--in comparison with the opposite wrist- but this was statistically significant only for ulnar deviation. The postoperative radiographic reductions were similar in both groups, with no differences in loss of reduction after bone healing. Therefore, in Colles' fractures, trans-styloid fixation with two K-wires seems to give a stable osteosynthesis, which does not need additional immobilization with a plaster cast. PMID- 10366910 TI - Fractures of the distal forearm in young adults. An epidemiologic description of 341 patients. AB - We describe the epidemiology of all distal radial fractures in young adults (men 20-59 years, women 20-49 years) in Lund (1992-95) and Malmo (1994-95), Sweden. During the study period, there were 341 patients with 346 fractures in the two cities, found through the Hospital Register of Diagnoses in Lund and the register of the Radiology Department in Malmo. More than half of the fractures were dislocated and 2/3 of the cases involved the radiocarpal or radioulnar joints, in contrast to the predominantly extra-articular fractures in the elderly. There was an even distribution between sexes and the fractures were mainly caused by a severe trauma, i.e., more than a simple fall, most often sports injuries in January, February and May. Our findings suggest that distal radial fractures in nonosteoporotic young adults should be regarded as a special entity, at least in epidemiological studies. Possibly they also require treatment differing from that for osteoporotic fractures. PMID- 10366911 TI - Forearm fractures in Malmo, Sweden. Changes in the incidence occurring during the 1950s, 1980s and 1990s. AB - Between the 1950s and the 1980s, the incidence of forearm fractures increased in the city of Malmo. We have now collected data on all forearm fractures during 1991 and 1992 and compared them with previously published data from 1953-1957 and 1980-1981. During the 1990s, 1314 individuals with wrist fractures and 125 with shaft fractures were recorded. In men, we found a twofold increase in the standardized morbidity ratio (SMR) in the 1990s, compared with the 1950s. The 1990s, compared with the 1980s, showed a reduction in SMR to 0.85. In women, a comparison between the 1990s and the 1950s revealed a slight reduction in SMR, 0.9 during the 1990s. Comparison of the 1990s with the 1980s revealed a reduction in SMR to 0.7 after the age of 70 years. In individuals 60 years and older, we found a fivefold increase in the incidence of fractures of the shaft of the forearm, when comparing the 1990s with the 1980s. In women, the increase in incidence of wrist fractures appears to have been interrupted, when comparing the years 1991-1992 and 1980-1981. Among men, the incidence of wrist fractures appears to be increasing, even after the 1980s. The reduction in incidence among women may partly be explained by warmer winters during 1991-1992. PMID- 10366912 TI - Intramedullary nailing of humeral shaft fractures. A retrospective study of 126 cases. AB - Antegrade intramedullary nailing with four different implants was used in 126 humeral shaft fractures. There were 74 acute fractures, 17 pathologic fractures, 16 fractures malaligned in a hanging cast or brace, 15 fractures with delayed union and 4 fractures that were nailed after failed open reduction and internal fixation. The nonunion rate was 21/95 after primary operation, and after reoperations 14/95. Distraction of the fracture was a significant cause of nonunion, but not type of fracture, localization, implant, and delay between injury and surgery. Shoulder joint function was significantly impaired in 25/67 patients. The patients regarded the result as good or satisfactory in 41/67 of the cases who were followed mean 3 (0.5-10) years. We conclude that antegrade intramedullary nailing of humeral shaft fractures leads to a substantial risk of non-union and impairment of shoulder function. It can be recommended as primary treatment only when nonoperative treatment is likely to fail. PMID- 10366913 TI - Neural response of mechanoreceptors to acute inflammation in the rotator cuff of the shoulder joint in rabbits. AB - We examined with electrophysiological techniques the effects of experimentally induced inflammation on the mechanosensitive afferent units in the rotator cuff of the shoulder joint of 21 rabbits. We identified 21 mechanosensitive units belonging to group III. 12 units had mechanical thresholds of > 7.0 g and 9 units had thresholds of < 7.0 g. After injection of inflammatory agents, kaolin and carrageenan, into the joint space, ongoing afferent discharge rates increased in all units. The average discharge rate increased significantly from 7 imp/s to 15 imp/s after injection. 5 units had a decreased mechanical threshold after the injection. Acute inflammation seems to have an excitatory and sensitizing effect on the high- and low-threshold units in the rotator cuff. PMID- 10366914 TI - Internally fixed femoral neck fractures. Early prediction of failure in 203 elderly patients with displaced fractures. AB - After internal fixation of a femoral neck fracture, 3 months is the critical time for planning rehabilitation of the patient. Most failures in the elderly occur within this time. In a series of 165 patients, we followed 127 women and 38 men with a median age of 81 (63-97) years from an examination at 3 months to reoperation or survival of the hip. 36 patients had radiographic signs of disturbed healing at the 3-month follow-up--change in fracture position by 10 mm, change in screw position by 5%, backing of the screws by 20 mm, or perforation of the femoral head by the screw. These signs had a high association with local complications and need for a later reoperation. High age and male sex increased this association. Signs of impaired healing made nonunion likely, but did not predict late segmental collapse of the femoral head. Patients with signs of disturbed healing and those closest to them should be informed about the value of early check-ups in case of pain and impaired function. PMID- 10366915 TI - Low IGF-I levels in hip fracture patients. A comparison of 20 coxarthrotic and 23 hip fracture patients. AB - We measured total body bone density and body composition with dual energy x-ray absorptiometry in 43 elderly patients, 23 with hip fracture and 20 with coxarthrosis, after surgery and after 6 months. Insulin-like growth factor-1 (IGF I), a polypeptide known to affect bone metabolism, and two of its binding proteins (IGFBP-1, IGFBP-3) were measured preoperatively and after 6 months. Normal serum IGF-I levels are dependent on adequate nutrition and normal secretion of growth hormone (GH). We found consistently lower levels of IGF-I and IGFBP-3 and a tendency to higher levels of IGFBP-1 in the patients with hip fractures, who also had a lower total body mass, lower fat mass and bone mineral density than the coxarthrosis group, indicating a more catabolic state in the patients with hip fracture, even 6 months after the trauma. PMID- 10366916 TI - Gene expressions of antiinflammatory mediators in THR retrieved interfacial membranes. AB - We investigated gene expression of antiinflammatory mediators in the interfacial membranes retrieved at hip revision arthroplasty using reverse transcription polymerase chain reaction (RT-PCR). Levels of RT-PCR products were compared with those of synovial tissue from patients with osteoarthrosis or rheumatoid arthritis. Antiinflammatory mediators such as type II interleukin (IL)-1 receptor, IL-4, IL-10, IL-1 receptor antagonist, and transforming growth factor beta 1 (TGF-beta 1) were expressed in the interfacial membrane. In interfacial tissue, the level of IL-10 was lower, but that of the IL-1 receptor antagonist higher than in diseased synovial tissue. PMID- 10366917 TI - Poor outcome of the PCA and Harris-Galante hip prostheses. Randomized study of 171 arthroplasties with 9-year follow-up. AB - 155 patients (171 hips) with a mean age of 50 years (24-64) were randomized to uncemented PCA (84 hips) or Harris-Galante type I (87 hips) total hip arthroplasty. Clinical and radiographic evaluations were done regularly. The improvements in the Harris hip and pain scores did not differ. Osteolysis developed in 5 PCA and 17 Harris-Galante hips. 13 hips in the PCA and 16 in the Harris-Galante (HG) group were revised because of mechanical failures and 1 hip (HG) because of infection after a mean follow-up of 9 years. Decreased 10-year survival rate, based on revision as end-point, was noted for the PCA (85%), compared with the Harris-Galante cup (99%). The corresponding survival rate of the PCA stem (96%) was higher than that observed for the Harris-Galante design (86%). When radiographic failures were included, the survival rates of the 4 different components dropped to between 73% and 94%. These findings indicate that further revisions will be necessary and continuous radiographic follow-up is indicated to enable revision before severe bone destruction has occurred. Although the PCA and the Harris-Galante designs differed as regards the survival of the individual components, the overall clinical and radiographic survival rates of these cementless total hip arthroplasties were poor. PMID- 10366918 TI - The economics of preventing revisions in total hip replacement. AB - We showed that the selection of a cost-effective type of cement and method of prophylaxis against deep infections for patients undergoing total hip replacement depended on the number of arthroplasties performed each year at individual hospitals. When 100 arthroplasties were performed each year, the use of Palacos cement and systemic antibiotics reduced the total costs to the department, i.e., the cost of cement, infection prophylaxis and revisions. The use of gentamicin impregnated cement in combination with systemic antibiotics will further reduce the risk of revision and is another cost-effective strategy. The most effective infection prophylaxis would be achieved with a combination of gentamicin impregnated cement, systemic antibiotics and surgical enclosure. However, the additional cost of the surgical enclosure would not be offset by cost savings due to reduced risk of revisions. PMID- 10366919 TI - Use of unicompartmental instead of tricompartmental prostheses for unicompartmental arthrosis in the knee is a cost-effective alternative. 15,437 primary tricompartmental prostheses were compared with 10,624 primary medial or lateral unicompartmental prostheses. AB - Unicompartmental knee arthroplasty (UKA) is known to have a higher risk of revision than tricompartmental arthroplasty (TKA), while UKA implants are generally less expensive than TKA implants. We estimated the costs of implants and hospital stay of both procedures and related the cost difference at primary operation to the difference in number of revisions to be expected. We compared 15,437 primary TKAs and 10,624 primary medial or lateral UKAs. The operations were all done on patients with arthrosis during 1985-1995. By matching patients in the Swedish Patient Administration System with the Swedish National Knee Arthroplasty Register, the groups could be compared regarding the length of the hospital stay. The cumulative revision rate (CRR) and the relative risk of revision were calculated with survival statistics, as well as the risk of a second revision and the risk of infection. The weighted mean cost of the commonest implants in each group was used as an estimate of the implant cost. We found that the TKA patients were, on average, 2 years older at operation and had a lower CRR than the UKA patients-i.e., 10-year CRR of 12% and 16%, respectively. After adjusting for age, gender and year of operation, UKA patients were found to have a 2-day shorter hospital stay and fewer serious complications than TKA patients. The mean estimated cost of a unicompartmental implant was 57% of that of a tricompartmental implant. We conclude, that by using UKA instead of TKA in appropriate patients, money can be saved, even after taking into account the increased number of revisions to be expected. PMID- 10366920 TI - Early changes in muscle strength after total knee arthroplasty. A 6-month follow up of 30 knees. AB - We studied 30 patients with arthrosis in one knee operated on with a cemented (n 26) or an uncemented total knee arthroplasty (TKA) (n 4). Full weight-bearing from the first postoperative day was allowed in all patients, and they received standard postoperative physiotherapy. 1 week prior to surgery, and after 3 and 6 months, isokinetic and isometric muscle strength in both legs were measured, using a Cybex 6000 dynamometer. Isokinetic tests showed a bilateral, significant, and progressive increase (30-53%) in flexor muscle strength most pronounced in the operated legs. Isokinetic extensor strength increased significantly (14-18%) in the operated legs, while in the contralateral legs, a limited increase was found. Isometric flexion strength significantly decreased in the operated knees (17%). Isometric extension strength showed a temporary decrease at 3 months, which returned to the preoperative level. No significant change in isometric strength was observed in the contralateral legs. The knee pain during the muscle strength measurements decreased significantly from the preoperative level, which may indicate that the substantial pain relief within 3 months after a TKA is an important factor for evaluation of muscle strength. PMID- 10366921 TI - Patellar tendon graft position after anterior cruciate ligament reconstruction. Interobserver variability on lateral radiographs. AB - We compared the reliability and validity of graft position measurements made by 4 orthopedic surgeons on intraoperative radiographs obtained using fluoroscopic control and postoperative radiographs obtained from the same 17 patients 6 weeks after ACL reconstruction. Measurements from postoperative radiographs varied significantly more than those from intraoperative radiographs. There was little agreement between the postoperative and intraoperative measurements of the tibial and femoral graft position. We conclude that postoperative radiographs are not a sufficient tool for assessing graft placement after ACL reconstruction using patellar tendon autografts. In order to consider graft position in follow-up studies and to compare results from various surgeons, we suggest intraoperative fluoroscopy to produce radiographs for accurate and reliable measurements. PMID- 10366922 TI - Anterior cruciate ligament reconstruction affects proprioception in the cat's knee. AB - To study the cat's knee after anterior cruciate ligament reconstruction, we compared its neural and muscular activity with that in the normal and the unstable knee. We recorded the electric activity in the articular nerves (posterior -PAN and medial -MAN) and periarticular muscles (quadriceps and hamstring) while performing passive flexion, extension, external and internal rotation, and also anterior translation of the tibia at 30 degrees and 90 degrees of flexion. The same series of maneuvers was performed in the same knees after surgical section of the anterior cruciate ligament and then after anterior cruciate reconstruction. The electric activity recorded in the reconstructed knee was compared to that in the same knee before surgery and in the same unstable knee after anterior cruciate section. We observed that the reconstructed knee, compared to the injured knee, showed a decrease in articular nerves and quadriceps activity while it regained stability. This decrease converged to the recordings in the normal knee. However, differences in MAN, PAN and hamstring activity were still present in the reconstructed knee. This suggests that, although anterior cruciate reconstruction seems beneficial for restoring articular nerve and periarticular muscle activities to a certain degree, proprioception in the reconstructed knee does not match that in the normal knee. PMID- 10366923 TI - Joint position sense is not changed after acute disruption of the anterior cruciate ligament. AB - We evaluated the impact of acute, isolated ACL disruption on knee joint proprioception by means of passive-active and active-active joint position sense (JPS) measurement techniques. 18 subjects with acute, isolated and unilateral ACL disruption were tested for JPS in a standing position. The test protocol included 6 trials for each leg. In each trial, the lower leg was passively positioned to an index angle approximating either 30 degrees or 70 degrees, followed by 5 active repetitions of the index angle where the subjects attempted to reproduce the index angle to the best of their ability. The errors from the exact index angle reproduction were calculated as both real (showing both magnitude and direction) and absolute values (only magnitude). All subjects had a tendency to reproduce the index angle with both the injured and normal knees in a more flexed position (overestimation). Only the absolute error produced by the active-active test at flexion angles greater than 45 degrees produced a significant difference with a larger error for the normal knee. In all other comparisons between the injured and the normal knee no differences were found. We conclude that the afferent signals which are compromised by an acute tear of the ACL are insignificant compared to afferent signals from the other joint and muscle receptors. PMID- 10366924 TI - Acute spinal epidural abscess without concurrent spondylodiscitis. Successful closed treatment in 10 cases. AB - We performed a retrospective survey of the clinical records and radiological examinations of 10 patients with a diagnosis of spinal epidural abscess, without spondylodiscitis. All patients had an acute onset of fever and local or radiating back pain. 3 patients had mild, and 1 patient severe neurological symptoms. The diagnosis and subsequent regression of the abscess after treatment were verified by MRI. In all cases, the imaging findings included signs of septic arthritis in an adjoining facet joint. 7/10 abscesses were located in the lumbar region. Blood cultures showed Staphylococcus aureus as the etiological agent in 8/10 patients. In 2 cases, no agent was found, probably due to ongoing antibiotic therapy when the cultures were taken. All patients were treated successfully using antibiotics alone, with complete regression of the neurological symptoms. PMID- 10366925 TI - Lumbar nerve root compression by intraspinal synovial cysts. Report of 8 cases. AB - We evaluated the clinical appearance and results of surgical treatment in 8 patients with leg symptoms due to a lumbar intraspinal synovial cyst. The most frequent symptom was radicular leg pain due to unilateral single-root compression affecting the L5 or S1 nerve root. There were 2 cases with large cysts causing compression of the cauda equina, with spinal claudication as the main symptom. All cysts arose from arthrotic facet joints. Surgical excision gave good results and no recurrences have been noted 0.5-2 years postoperatively. PMID- 10366926 TI - Reduced reperfusion injury in muscle. A comparison of the timing of EPC-K1 administration in rats. AB - EPC-K1, a phosphate diester of alpha-tocopherol and ascorbic acid, is a new hydroxyl radical scavenger. We examined the effects of EPC-K1 according to differences in the timing of its administration. Warm ischemia, produced by vascular pedicle clamping, was sustained for 4 hours. After 24 hours of reperfusion, muscle injury was evaluated in 4 groups: the first group received a sham operation, the second group was treated with an intravenous injection of EPC K1 prior to ischemia, the third group was treated with EPC-K1 prior to reperfusion, and the fourth group was controls. Compared with the control group, both the preischemic and pre-reperfusion EPC-K1-treated groups showed a statistically significant amelioration in the reduction of isometric muscle contraction. There were also significant reductions in the muscle and serum levels of thiobarbituric acid reactive substances (TBA-RS) and muscle damage, indicated by the biochemical and histological study. A comparison of the timing of EPC-K1 administration revealed that only the muscle TBA-RS level in the pre reperfusion EPC-K1-treated group was significantly higher than that in the preischemic EPC-K1-treated group. These observations indicate that EPC-K1 not only by preischemic but also by pre-reperfusion administration acted effectively on reperfusion injury in muscle, thereby improving muscle function. PMID- 10366927 TI - Ethanol and its effects on fracture healing and bone mass in male rats. AB - Operatively induced, standardized tibia fractures in 42 10-week-old male rats were fixed with intramedullary nails. 21 of the rats were fed liquid containing 15% ethanol. 5 weeks after inducing the fracture, the rats were killed and the total body bone mineral density (BMD) was analyzed with the DEXA technique, and the mechanical properties of the fractured and the unfractured tibiae as well as the ipsi- and contralateral femoral shaft and femoral neck were tested. The rats given a liquid containing 15% ethanol were found to have significantly lower total BMD and total calcium than the controls. We also found a significantly lower bending moment and bending stiffness both in the fractured and unfractured tibiae among rats fed on ethanol. The energy absorption until refracture was less in rats fed on ethanol. Posttraumatic osteopenia was present, as judged by the mechanical tests of the ipsilateral femoral shaft and the femoral neck in all animals. There was no difference in this respect between the animals fed on ethanol and the controls. We found that ethanol disturbs bone metabolism which reduces the mechanical properties of the tibiae and femora of rats, but the healing process of an induced tibial shaft fracture was not affected. PMID- 10366928 TI - Hydroxyapatite coated with insulin-like growth factor 1 (IGF1) stimulates human osteoblast activity in vitro. AB - We studied the effects of a hydroxyapatite-tricalcium phosphate material coated with Insulin-like Growth Factor 1 (IGF1) on cell growth, collagen synthesis and alkaline phosphatase activity (ALP) of human osteoblasts in vitro. Cell proliferation was stimulated when osteoblasts were incubated with untreated hydroxyapatite (HA) and it was further increased by exposure to IGF1-coated HA. 3H-Proline uptake was significantly increased by treatment with either HA or IGF1 coated HA but no significant differences were found between these two groups. ALP activity was enhanced by exposure to HA, with respect to the control, and further increased by treatment with IGF1-coated HA. Our findings suggest that HA is useful for promoting osteoblast activity and IGF1 may help to improve its biological characteristics. PMID- 10366929 TI - [Acute self-induced poisoning with carbamazepine]. AB - OBJECTIVE: We report the epidemiological, clinical, toxicological and therapeutic aspects of acute voluntary intoxication with carbamazepine. PATIENTS AND METHODS: The study included 17 cases of acute carbamazepine intoxication in patients hospitalized in our toxicology unit. RESULTS: Neurological signs predominated at admission, mainly agitation or coma associated with seizures. Mydriasis and cardiovascular signs were frequent. Blood chemistry most frequently showed hyponatremia. Mean serum carbamazepine level admission was 24 mg/l (range 4 n 12 mg/l). Ten patient required respiratory assistance for 28 +/- 17 hours. Symptomatic treatment and gastric lavage (+activated carbon) provided favorable outcome. CONCLUSION: Acute carbamazepine intoxication is seen with increasing frequency. Severity is related to the degree and duration of the coma, respiratory depression, seizures, cardiovascular disorders, and metabolic abnormalities. Symptomatic and specific treatment with activated carbon are required. PMID- 10366930 TI - [Visceral leishmaniasis associated with Wegener disease. Use of lipid complex amphotericin B and liposomal amphotericin B]. AB - BACKGROUND: Leishmaniasis in a patient with Wegeneris disease raises the problem of amphotericin toxicity further compromising the pre-existing renal disorder. CASE REPORT: An anemic patient treated for Wegeneris disease developed visceral leishmaniasis. This renal failure patient was treated with lipid complex amphotericin B and liposomal amphotericin B. We report outcome at 10 months follow-up. DISCUSSION: The new formulations of amphotericin B allow effective treatment of visceral leishmaniasis in renal failure patients. Long-lasting results are probably favored by the interruption of immuno-suppressive therapy. PMID- 10366932 TI - [Fatal posttransfusion Enterobacter amnigenus septicemia]. PMID- 10366931 TI - [Mitral papillary fibroelastoma in a HIV infected patient]. AB - BACKGROUND: Papillary fibroelastoma of the endocardium is a benign cardiac tumor which may cause embolic events. CASE REPORT: We report the first case, to our knowledge, of cardiac papillary fibroelastoma which was revealed by cerebral ischemic events in a patient infected by the human immunodeficiency virus. The initial diagnosis was mitral endocarditis complicated by cerebral embolic events. DISCUSSION: This case demonstrates that fibroelastomas should be added to other valvular diseases described in HIV infected patients. We recall the embolic potential of cardiac papillary fibroelastomas which warrant surgical treatment. PMID- 10366933 TI - [Acute hepatitis after phytotherapy]. PMID- 10366934 TI - [Apropos of treatment of lead poisoning]. PMID- 10366935 TI - [Iatrogenic nosocomial drug-induced conditions: where is the risk?]. PMID- 10366936 TI - [Extemporaneous anatomo-pathologic examination of breast and thyroid diseases]. PMID- 10366937 TI - [Apropos of extemporaneous anatomo-pathologic examination of thyroid diseases]. PMID- 10366938 TI - [Apropos of anatomo-pathologic examination of breast diseases]. PMID- 10366939 TI - [Carcinomatous lymphangitis]. AB - DEFINITION: Carcinomatous lymphangitis is a radioclinical entity accounting for about 8% of all cases of lung metastasis defined as the presence of tumoral cells in lymph vessels and lung interstitium. DIAGNOSIS: Biopsy specimens or bronchial brushings obtained by fibroendoscopy or bronchioalveolar lavage fluid usually reveal adenocarcinoma. PRACTICAL MANAGEMENT: In clinical practice, the patient presents with dyspnea and non-specific infiltration on the chest x-ray. The clinical situation worsens rapidly. Millimetric CT-scan shows highly suggestive polygonal images in the subpleural area. Respiratory function tests may be helpful for the differential diagnosis, particularly in difficult cases, showing a mixed ventilation disorder without altered carbon monoxide diffusion and hypoxemia at rest without hypercapnia. SEARCH FOR THE PRIMARY CANCER: Primary lesions must be identified for specific treatment. Pathology findings help guide the search. Despite the highly unfavorable prognosis (median survival = 3 months), etiological treatment when possible can improve quality of life and possibly survival. Symptomatic treatment is indicated and must be adapted to each individual case. PMID- 10366940 TI - [LDL-apheresis therapy, methods and indications]. AB - A SEVERE DISEASE: Homozygote hyper cholestrolemia is a very severe disease with an extremely high atherogenic potential. The risk of sudden death starts at about the age of 10 years and mortality reaches nearly 100% at 20 years. In this homozygote autosomal dominant disorder, serum cholesterol rises above 6 g/l due to a total deficiency of LDL-receptors. LDL-APHERESIS: The treatment of choice, LDL-apheresis, is an aggressive treatment. Drug regimens and palliative surgery have little or no effect. Liver transplantation is highly effective but compromises long term prognosis. LDL-apheresis, indicated in case of drug resistance, can be used to clear atherogenic LDL particles extracorporally. Currently it is the most effective means of lowering serum cholesterol in these patients and avoid potential cardiovascular complications. SEVERAL TECHNIQUES: The most specific techniques are also the most costly. These techniques allow treating whole blood or previously separated plasma. Such specific techniques are indicated in children. Compared with other European countries such as Germany, the development of these techniques has been retarded in France due to the lack of financing by the national health insurance system. PMID- 10366941 TI - [Screening for exogenous erythropoietin]. AB - r-HuEPO: Ever since it was first produced, illicit use recombinant human erythropoietin (r-HuEPO), a human polypeptide which accelerates production and maturation of red cells and consequently improves aerobic potential, has been observed in athletic competitions. IMPORTANCE OF SCREENING: Unfortunately, abuse of r-HuEPO can have severe adverse effects, particularly on the cardiovascular system. Screening methods capable of detecting exogenous erythropoietin were thus developed not only to detect illicit drug use in sports competition, but also to preserve athletesi health. THEORETICAL POSSIBILITIES: Several methods have been explored. Direct methods using electrophoresis or indirect methods measuring different biological parameters (fibrin degradation products, number of soluble transferrin receptors) as well specific hematology tests (red cell morphology, hematocrit...) can theoretically detect exogenous erythropoietin. METHODS TO VALIDATE: It would be advisable to examine all of the available work on the proposed methods to develop a reliable diagnostic method for detecting r-HuEPO. Soluble transferrin receptor counts appear to be the most promising method, but specificity is not totally satisfactory. PMID- 10366942 TI - [Current problems of chemical radiation protection of organisms]. AB - The classification has been proposed for antiradiation protective agents which are divided into four groups: I. Radioprotectors; II. Adaptogens; III. Absorbents; IV. The means of Rehabilitations. Radioprotectors in turn are subdivided into myelo-, entero- and cerebroprotectors. Adaptogens act as stimulators of radioresistance. These are natural protectors which are perspective for chemical protection under low-level ionizing irradiation. Natural protectors influence regulatory systems of exposed organisms, mobilize endogenous background of radioresistance (EBR), immunity and intensivity the total non specific resistance of organism (TNRO). Natural protectors (extracted from cells, plants, animals) are low- or nontoxic and can be used with food. Absorbents, the means of protection from internal irradiation, are subdivided into the drugs which prevents incorporation of radioiodine by thyroid gland and absorption of radionuclides (137Cs, 90Sr, 239Pu, 241Am) in the digestive tract. The main features which distinguish radioprotective drugs of different mechanisms on human organisms are presented. The main which different programs of reability on invalids-Chernobyl. PMID- 10366943 TI - [Classification of radiation protective agents as a basis of modern radiation pharmacology]. AB - The consistency of the classification of prophylactic antiradiation drugs have been given consideration as history of their discovery, theory of the radioprotection mechanisms and their use in applied medicine. To prophylactic antiradiation drugs can be related to the drugs of having capable of decreasing radiation injuries burden while administrated before or immediately after radiation. The ones consists of radioprotectors with short-term of long-term action, drugs stimulating radioresistance, the ones suppressing symptoms of primary radiation reaction, the ones of early detoxication, the ones for absorbtion and elimination of radionuclides from an organism. PMID- 10366944 TI - [Radiation protective agents effective in a wide range of radiation dosages]. AB - Perspective chemical classes of prophylactic radioprotecting compounds were found from analysis of proper experimental data. Preparations effective in a wide range of radiation doses of various intensity were synthesized. Their activity are due to combination of antiradiation and many other pharmacological properties. During acute radiation these preparations protect stem cells pul of critical tissues but under prolonged radiation they normalize the hematological, immune and lipid peroxide oxidation systems. There is no strong correlation between radioprotection and increasing of functional reserve of organism. PMID- 10366945 TI - [Strategy and tactics of radiation protection of the personnel in light of future work on stabilization and reorganization of the "Shelter" compound into radiologically safe system]. AB - The research aim is to analyze the "Shelter" personnel work conditions and to estimate how these conditions specify radiological defense. "Shelter" object remains to be an accumulation of the open radiotoxic materials. It is expected that the usage of standard technologies in such conditions will be connected with the great dose-expense of external and internal irradiation. The significant quantity of the fuel dust (more 30 tonne according to uranium) and the extremely harmful microclimate conditions such as: aggressive radiochemical aerosols, exclusively artificial and obviously insufficient illumination, high humidity, discomfortable temperature regime, high ability of many working places to induce the emotional stress reaction in personnel are the specificity of "Shelter" object. The synergetic influence of radiofactor and "Shelter" object microclimate is very dangerous because of its ability to deteriorate significantly the personnel health up to acute stress reaction. In these cases there appear the episodes of the short disorders of orientation in the surrounding situation, paroxysmal consciousness disturbances which may become the nondirect reason of the overirradiation and growth of work traumatism. That's why the methods of the stress-reaction prophylaxis and improvement of the individual defense become extremely improvement for the "Shelter" object personnel. It was shown that the following medicines are perspective due to their adaptogenic and radioprotective abilities, such as: alpha-2-recombinant interferon, antiinflammatory, steroid, antioxidant and enterosorbtive preparations of biological nature. PMID- 10366946 TI - [The role of cell hypoxia in the effect of radiation protectors]. AB - In experiment with mice, rats and dogs the intimate relationship has been established between radioprotective efficiency of indraline, cystamine and mexamine and its properties to enhance succinate dehydrogenase (SDG) activity of blood lymphocytes (r = 0.95). In that analysis modifying effect of normobaric hyperoxia has been estimated. Mice and dogs were correspondingly irradiated with gamma-60Co-rays in the dose 8.33 and 3.16 Gy. In the investigation involving mice, dogs and men the effect-dose dependence of aggravating of the SDG activity was linear for indraline, but not for cystamine. In man hypoxic hypoxia with air hypoxic mixture containing 10% of oxygen has initiated rise of the SDG activity being twice as smaller as the one when indraline in the dose of 100 mg administrated. Hyperoxia suppressed radioprotective properties of indraline, cystamine and mexamine in the ED50 in term of DRF by twice and didn't virtually influenced on that in the optimum radioprotective doses. Hyperoxia and alpha adrenoblocator tropaphene also suppressed the SDG response to indraline. In vitro experiment cystamine and adrenaline held stimulating action on SDG of blood lymphocytes. The role of pharmacological stimulation of cell respiration and cell hypoxia relating with the one in mechanism of radioprotective effect of the radioprotector of two dissimilar groups was discussed. PMID- 10366947 TI - [Radiation protective efficacy of alpha-adrenomimetics during local gamma irradiation of the skin]. AB - In experiment with mice and rats radioprotective properties of direct alpha adrenomimetic drugs: indraline, mesaton (phenylephrine) and naphthyzine (naphazoline) have been investigated by local early and late radiation injuries. The drugs were S. C. and C. administrated at the site of the thigh of the hind of the animal at intervals of 5-8 min locally was irradiated with gamma-60Co-rays in the doses of 26.3-53.7 Gy at a rate of the dose of 1.31-1.54 Gy/min. When S. C. injected the topical effect of the adrenomimetics was higher than the systemic one. In experiment with mice radioprotective efficiency of topical indraline (S. C. 50 and 100 mg/kg) in term of DRF was equal to 1.34 and 1.67 for radiation burn of the skin, 1.56 and 1.91 for 3rd month radiation contracture of the hind and 1.10 and 1.50 for partial amputation of the one. In the same condition the effect of topical mesaton (S. C. 1, 2.5, 5 mg/kg) in term of DRF was accordingly equal to 1.29, 1.49 and 1.62 for early radiation injury, 1.08, 1.20 and 1.46 for 3th month radiation contracture of the hind and 1.0, 1.14, 1.33 for partial amputation of the one; the effect of topical naphthyzine (5 mg/kg) in the term of DRF was 1.36 for early radiation injury. In experiment with rats radioprotective property of systemic indraline (I. M. 100 mg/kg) in the term of DRF was equal to 1.39 for radiation burn of the skin, 1.39 for 6th month radiation contracture of the hind, 1.41 for partial amputation of the one. When C. administrated the effect of topical indraline (5% solution in 50% DMSO or ethyl alcohol solution) in term of DRF was accordingly equal to 1.17 and 1.18 for radiation burn ot the skin, 1.50 and 1.35 for radiation contracture of the hind, 1.41 and 1.28 for amputation of the one. When S. C. administrated the effect of topical and systemic mesaton (2.5 mg/kg) in term of DRF was accordingly equal to 1.30 and 1.04 for radiation burn ot the skin. 1.25 and 1.0 for radiation contracture of the hind, 1.30 and 1.0 for amputation of the one. PMID- 10366948 TI - [Toxicity and radiation protective properties of 2(3)-guanidinoalkanethiols: structure-activity]. AB - In experiments with mice radioprotective properties and toxicity of 2(3) guanidinoalkanethiols were investigated by exposure to gamma-irradiation in the dose of 6.4 Gy. The obtained results allow to find an association between radioprotective activity of 2(3)-guanidinoalkanethiol compounds and their structure and toxicity. PMID- 10366949 TI - [Increased efficacy of radiation protection against fission neutrons using unithiol]. AB - It was found that the combination of unithiol (Sodium salt of 2,3-dimercapto-1 propansulfonic acid) with cystamine and AET diminished their toxicity. The optimum ratio for the antitoxic effect is 0.5 molar equivalent of unithiol per radioprotective 1.0 equivalent of thiol. Animals withstand big doses of protectors well, that gives an opportunity to use increased amounts of cystamine and AET. In the experiments with circular irradiation of male (CBA x C57B1)F1 mice weighing 18-22 g with fission neutrons (the neutron mean energy was 0.85 MeV, the contribution of gamma-quanta to the total was 25%, dose rate was 14 cGy/min) it was shown that the combination of unithiol with cystamine and AET enhances their radioprotective effect: the DRF of cystamine (150 mg/kg)--1.1, and the DRF of the combination of cystamine (300 mg/kg) with unithiol (152 mg/kg)- 1.2; the DRF of AET (150 mg/kg)--1.2, the DRF of the combination of AET (300 mg/kg) with unithiol--1.4. Thus, the enhancement of dose of the radioprotectors, which was made possible as a result of their combination with unithiol, leads to enhancement of efficacy of chemical protection against fission neutron irradiation as much as 10-20%. Efficacy of AET is found to be comparable to efficacy of this protector in conditions of X-rays irradiation. PMID- 10366950 TI - [Effect of unithiol on cystamine toxicity in dogs]. AB - In experiments with dogs it was shown that administration of unithiol before administration of cystamine decreases toxic effect of the last: period of excitation induced by cystamine is shortened, the time of emetic reaction decreases and expression of the emetic reaction to this preparation is diminished, the recorded EKG changes of conditions of heart decrease. PMID- 10366951 TI - [Radioprotective effect of lymphokinine]. AB - It is shown that lymphokinine, cytokine complex, i. a. applied in mice (50,000 U/kg) 24 hours before the following acute of prolonged irradiation in dose LD50 85/30 increases animal survival by 30-40%. Lymphokinine shifts down the expression of postradiational leucopenia, significantly accelerates leucocytes and myelokaryocyte recovery and stimulates colony-forming ability of bone marrow cells in irradiated animals. Lymphokinine induced increase of endogenous radioresistance rate is suggested to be associated with its ability to accelerate early haemopoietic cells proliferative and migration activity and modify cells cycle of marrow haemopoietic cells. PMID- 10366952 TI - [Radioprotective properties of carnosine]. AB - In experiments with rats and mice there was shown the ability of carnosine (beta alanyl-L-histidine) to depress accumulation of lipoperoxidative products in blood serum and to increase the yield of spleen endocolonies in irradiated animals. There were investigated duration of the protective action and compared effects of single and prolonged administration of carnosine. Carnozine restores the postradiative decrease in the cytochrome P-450 content in rat liver microsomes. Besides, the ability of carnosine to prevent the postradiative decline in the activity of UDP-glucoronyltransferase. Some possible mechanisms of its radioprotective action are discussed. PMID- 10366954 TI - [Evaluation of radioprotective properties of a lipocarotenoid extract from mycelium of basidiomycetes during external irradiation]. AB - Radioprotective properties of LPC extract from mycelium of Basidiomycetes fungus at doses 12.0; 3.0 and 1.0 Gy of external irradiation were investigated. The extract of LPC at 2.5 mkg/ml concentration promoted the survival of animals (to 40%) and increased the average life at the lethal dose. At 1.0 Gy dose the use of extract resulted in restoration of radiosensitive organs weights, the number of blood leucocytes and some indexes of lipid peroxidation in blood plasma. The extract has produced some effect on the amount of spermatogenic cells in the testis. PMID- 10366953 TI - [Increase in radiation resistance of an organism after administration of metabolic drug MR-33]. AB - Original MP-33 metabolic drug has inductive action on intracellular levels of glutathione (GSH) and GSH-related enzymes (glutathione peroxidase-GPx, glutathione S-transferase-GST) of animals and human (liver, heart, erythrocytes). Treatment of Wistar male rats with MP-33 per os (6 mg/kg) 30 min before gamma irradiation with a dose of 7 Gy results in 35% survival and increasing of mean life. In "mother-foetus" system the induction of intracellular level of GSH, GST and GPx by MP-33 leads to enhancement of foetus radioresistance. In contrast to MP-33 us of GSH (6 and 100 mg/kg) is not effective. Perspective of MP-33 us in relation to radioecological crisis is discussed. PMID- 10366955 TI - [Inhibition of lipoxygenase activity reduces radiation-induced DNA fragmentation in lymphocytes]. AB - Lypoxygenase activity of lymphocytes towards exogenous substrate linolic acid and the level of endogenous arachidonate metabolite 15-HETE are increased within 6 h and decreased below the control level at 24 h after 1 Gy X-ray irradiation of Wistar rats. DNA fragmentation is stimulated both in vivo within 12 h and during in vitro incubation of lymphocytes, obtained 3 h following irradiation. DNA analysis by agarose gel shows that DNA fragments are multiples of the 200 base pair unit. When 20 microM nordihydroguayaretic acid, a selective inhibitor of arachidonate lypooxidation, was added to the incubation medium, DNA fragmentation was observed to be significantly less, especially at the early steps of incubation. PMID- 10366956 TI - [Therapeutic effect of ultraviolet irradiation of autologous blood in early period of acute radiation sickness]. AB - In experiments with dogs the acute radiation sickness was caused by common relatively uniform 3.5 Gy dose gamma-irradiation. Reinfusion of UV-irradiated autologous blood induced undoubted curative effect, which manifested itself in increased mean life, reduced mortality, more full restoring of blood-formation. PMID- 10366957 TI - [Efficacy of radioprotectors and gene mutations following 2-deoxyribosyl damage. Computer modeling]. AB - The fundamental new universal method of evaluation of the interaction between macroradical and radioprotector (the access window method) is presented here. It's based on the modelling phenomenon of molecule penetration to the active centre of macromolecule structure through the functional groups free "window". The steric modelling of the B-DNA structure fragments allowed to measure the conformation parameters of the intermediate stereocomplex between interacting substancies. Using the thesis about molecule InH mask the interaction process of InH (steric hindered phenol 4-oxy-3,5-di-tret-butyl-alpha-metylbenzylamine, mercaptoethanol, MET, and L-cysteine, CYS) with 2-deoxyribosyl five radicals in DNA was studied. It was determined, that the radioprotection maximum effect on single-strand breakage formation in irradiated cell can reach 2/3 of the total sugar radicals yield and for MET and CYS (with minor mask squares) -87.5, and 91%, accordingly (it agrees with experiment in literature data). PMID- 10366958 TI - [Prevention of atherosclerosis and myocardial infarction in the liquidators of the accident at the Chernobyl Power Plant]. AB - The examination of 1450 patients-participants of elimination consequences after Chernobyl accident has revealed reasons of disablement: mental amd neurologic diseases -32%; cardiologic -28%; oncologic -13%; traumatic -14%. The reason of death in 60% were cardiologic diseases. Arteriosclerosis and myocardial infarction were revealed among participants of 1986 in 85% and 1987 only in 15%. There were revealed high efficiency of myocardial infarction's prophylactic by means of prescription hypoholesterinemic preparations. PMID- 10366959 TI - [Characteristics of infection in children during prolonged oxidative stress after radiation exposure]. AB - In the article on discusses the problems of origin and duration of chlamydia and herpes-virus infection in children with chronic somatic pathology and different degrees of radiation stress in contaminated territories of Brianskaya region (the level of contamination according to 137Cs = 9.37-19.73 Ci/km2). PMID- 10366960 TI - [Adaptogen MIGI-K in rehabilitation of Chernobyl liquidators]. AB - Experimental studies of the mussels hydrolysate (MIGI-K) have shown that this preparation can enhance the radioresistance and general resistance of the organism. The mechanisms of its action are related mainly with its influence on the components of the biochemical back-ground of the resistance and with its antioxidative activity. Some properties of this preparation, for example, absence of toxicity and harmful side effects, made it possible as an adaptogene and as a remedy, increasing radionuclides excretion. PMID- 10366961 TI - [Effect of mussels hydrolysate MIGI-K on the adaptive response od bone marrow hematopoietic stem cells in mice]. AB - Radioprotective action of MIGI-K hydrolysate on adaptive response was studied. It was shown that this preparation protecting animals under lethal and sub-lethal doses of irradiation can also modify adaptive response. It was supposed that adaptive response model can be applied for studying possibilities of radioprotectors' use under low doses irradiation. PMID- 10366962 TI - [Protective effects of MIGI-K during UV irradiation of animals]. AB - Prophylactic peroral use of MIGI-K preparation (products of acid hydrolysis of mussels meat) largely or completely prevented intensification of lipoperoxidation and depression of antioxidative systems (superoxide dismutase, glutathione peroxidase, nonprotein thiols, lipoantioxidants) in skin and liver of UV irradiated rats. PMID- 10366963 TI - [Efficacy of antioxidant preparations used for correction of impairment of oxidative homeostasis in Chernobyl liquidators]. AB - The influence of two antioxidative preparations (Vetoron, Morevit) on the parameters of antioxidative system and lipoperoxidative processes in blood of men, who took part in the cleaning up after Chernobyl NPP accident, under the clinical control in the hospital of Scientific Center for Radiation Medicine AMS of Ukraine was studied. It was shown that Morevit (extract of protein hydrocarboneus concentrate of Mytilis galloprovincialis) is more effective as promoter of normalization lipoperoxidative processes disorders and restoration of antioxidative enzymes activity. The main principles of antioxidative therapy were formulated. PMID- 10366964 TI - [Radioprotective properties of phenoxane during low intensity low dose gamma irradiation]. AB - The effect of prophylactic and therapeutic administration of phenoxane on the parameters of lipid peroxidation (LPO) regulatory system in murine tissues under low intensity gamma-irradiation (15 cGy) over a wide range of the agent doses is studied in dynamics. The different pattern of the phenoxane effect depending on its dose as well as the time of administration (before or after irradiation) is revealed with physiological tests (the dynamics of leucocyte amount, the blood formula, the change in spleen mass per mouse), on biochemical level (the changes in the LPO products content in murine blood and tissues), in biophysical properties (the lipid antioxidant activity level, the rate of oxygen consumption under the initiated oxidation of ethylbenzene in the presence of lipids from the intact and experimental murine organs) and also by the analysis of variability of the biochemical and biophysical properties in tissues and blood leucocytic formula within groups of mice. PMID- 10366965 TI - [Effect of melanin on mutagenic activity of chronic irradiation and adaptive response in mice]. AB - Pigment melanin peroral injection was shown to decrease significantly mutagenic action of chronic irradiation at low dose rate (0.007 Gy/h), and melanin protection against chronic irradiation is even more effective than against acute one. A phenomenon of radioadaptive response was revealed in mice in vivo by tests of chromosome aberrations in bone marrow cells and reciprocal translocations in germ cells. Adaptive response before priming dose was not observed if melanin was injected. If melanin was applied between conditioning and basic doses both radioprotective and adaptive effects were observed. PMID- 10366966 TI - [The use of nontraditional approaches to the analysis of the effects of radiation on the system of protein phosphorylation in spleen lymphocytes]. AB - Postradiation disturbances in functioning of Ca(2+)- and cyclic nucleotide dependent protein kinase systems are established. Kinetic index of different protein kinases of spleen lymphocytes under irradiation with doses of 0.5 and 1 Gy are analysed using return-cards-construction and Lyapunov's-coefficient determination methods. It is shown that determined disturbances can be considered as a part of general unspecific adaptational reaction of lymphocytes to irradiation. PMID- 10366967 TI - [Effect of beta-carotene oil and bee pollen on ion transport in rat brain slices following radiation-chemical exposure]. AB - Chronic combined exposure to ionizing radiation with dose of 0.25 Gy and cadmium chloride or atrazine to be present in drinking water at five-fold Limited Permissible Concentration (LPC) values led to the additively reduced intercellular K+ level in rat brain, that was at first choice caused by the active ion transport disorders in the case of irradiation, and by the changes in membrane permeability in the case of toxic loading. Applying of beta-carotene oil and bee pollen both abolished radiation effects, but no chemical toxicant ones. Authors supposed the selective action of the observed drugs to be connected with antioxidant activities of them. PMID- 10366968 TI - [Possible modification of radiation injury using radio frequency electromagnetic radiation]. AB - The possibility of radioprotective action of electromagnetic fields and radiations in radiofrequency range have been considered. It has been shown that the EMF and EMR effects depend on parameters of acting field. It is necessary to establish biophysical and biochemical ways and mechanisms of EMF and EMR action for effective use of radioemissions as radioprotectors. PMID- 10366969 TI - [Use of metal salts for radioprotection of plants during radioactive pollution of the territory]. AB - The applying salts of some metals to radionuclide contaminated soddy-podzolic soil in the zone of Chernobyl nuclear power station or the spraying of plants by its solutions are showing the radioprotective effect (salts of iron, zinc, cobalt and manganese) and decreasing the uptake of 90Sr and 137Cs through roots (salts of zinc, manganese, boron, lithium, cobalt and copper). PMID- 10366970 TI - [Radiation activation of angiotensin-converting enzyme during low-dose irradiation]. AB - Radiation activation of angiotensin-converting enzyme (respect to Cbz-Phe-His-Leu as substrate) was obtained at the gamma (137Cs, t(irr) = 10s-2h, D approximately 3 Gy)- and X (plasma foces source, t(irr) = 10(-9)s, Cu-filter, D approximately 2 x 10(-5) Gy)-irradiation. The inactivation of the horseradish peroxidase at the same X-irradiation dose (2 x 10(-5) Gy) took place. Based on the experimental data and on the mathematical model proposed by us we made a conclusion that the special points exist on the dose response curves. Besides, the deduction that an activation is a common process at radiation changes of the different enzymes follows from our model. The activation of tobacco peroxidase (in presence of Ca(2+)-cations and without them) and of recombinant horseradish peroxidase (in presence of H2O2) by gamma-irradiation was really observed (respect to guaiacol as substrate). PMID- 10366971 TI - [Humoral and cell mediated immunity induced by HBV-S gene recombinant retroviral vector]. AB - OBJECTIVE: To investigate the effectiveness of recombined retrovirus vector in gene therapy. METHODS: The retroviral vector PLXSN-S was constructed and transferred into PA317 by means of electroporation, then HepG2, P815, and EL4 cells were infected with the pseudovirus produced from PA317. In the meantime the gene immunization of PLXSN-S was also studied. RESULTS: HBsAg was expressed variously in the eukaryotic cells mentioned above and successfully evoked antibody response and CTL response to HBsAg after gene immunization. CONCLUSION: Intramuscular DNA immunization represents an efficient technique for priming CTL and antibody response to HBsAg. PLXSN may be used as an effective vector to carry genes of interest to target cells and immunization with it's DNA vaccine might be clinically useful for gene therapy. PMID- 10366972 TI - [Detection of NV-HB/s DNA and c-myc mRNA in mice inoculated with hepatitis B nucleic acid vaccine--NV-HB/s]. AB - OBJECTIVE: To explore the persistence period of plasmid in mice muscles injected with NV-HB/s and the possible influence on oncogene C-myc. METHODS: Anti-HBs was detected by ELISA; NV-HB/s was detected by In-situ and Southern-blot hybridization separately; C-myc mRNA was detected by In-situ hybridization. RESULTS: All mice injected with NV-HB/s were positive of anti-HBs after 1 month of immunization; NV-HB/s was negative when detected by In-situ hybridization; By Southern-blot hybridization, the positive rates of NV-HB/s were decreasing from 100% in 2 months to 25% after 6 months of inoculation. CONCLUSIONS: NV-HB/s would be disintegrated little by little after injected into mouse muscles; no C-myc activation was observed in 6 months after NV-HB/s inoculation. PMID- 10366973 TI - [Inhibitory effect of triplex forming oligodeoxynucleotides on HBV replication and synthesis of antigen]. AB - OBJECTIVE: To explore the inhibitory effect of triplex forming oligodeoxynucleotides(TFO) on replication of HBV and synthesis of antigen. METHODS: A 21 mer phosphorothioate triplex forming oligodeoxynucleotides (TFO21) directed at SP1 sites in HBV core promoter and a control of 21 mer phosphorothioate oligodeoxynucleotides (ODNcon) were synthesized. HepG 2.2.15 cells which can produce HBsAg, HBeAg, HBV DNA and HBV particle were treated with TFO21 and ODNcon. In HepG 2.2.15 cells treated with these oligodeoxynucleotides, the levels of HBsAg, HBeAg, and HBV DNA were examined by ELISA and dot hybridization method, respectively. RESULTS: The levels of HBsAg, HBeAg and HBV DNA in TFO21 group were lower than those in the bank control group. At concentration of 10 mumol/L, TFO21 inhibited synthesis of HBsAg and HBeAg by 57.5% and 77%. The inhibitory effect of TFO21 was dosage-dependent, and was related to time in which 2.2.15 cells were incubated with TFO21. No inhibition was observed in the ODNcon group. No toxicity was observed in the 2.2.15 cells treated with oligodeoxynucleotides. CONCLUSION: These results indicate that TFO is a potent inhibitor for HBV replication and synthesis of antigen, and also suggest that TFO is a therapeutic potential for the treatment of patients infected with HBV. PMID- 10366974 TI - [Study of transcription and cleavage in vitro of HDV with HBV-specific hammerhead ribozyme]. AB - OBJECTIVE: To study the effect of hepatitis B virus(HBV) specific ribozyme(RZ) and recombinant hepatitis D virus(HDV) inserting hammerhead ribozyme(rHDVRZP and rHDVRZA). METHODS: 831 bp HBV C gene fragment was cloned under the control of T7 promoter, 32P-labeled HBV transcript was incubated with gel-purified RZ, rHDVRZA, rHDVRZP at different temperature and autoradiographed after denaturing gel electrophoresis. RESULT: These results show that rRZ, rHDVRZA, rHDVRZP were active at 37 degrees C and more so at higher temperatures. CONCLUSION: Recombinant Delt virus could serve as a vector for the delivery of a ribozyme specific for hepatitis B virus cleavage. Our data demonstrate the value of recombinant ribozyme as potential therapeutic agents for treatment of HBV infection. Further study about cleavage in vitro and in vivo will continue. PMID- 10366975 TI - [Inhibitory effect of replication and expression of HDV by antisense oligodeoxynucleotides in H1 delta 9 cell]. AB - OBJECTIVE: To study inhibitory effect of an antisense oligodeoxynucleotide (ASODN) and its phosphorothioate (S-ASODN) on replication and expression of HDV in H1 delta 9 cell. METHODS: In previous studies, it was proved that the ASODNs which are complementary to genomic HDV ribozyme self-cleavage site and stem I regions can inhibit availably genomic HDV ribozyme activity. In the present study, a 15-mer ASODN and S-ASODS which are complementary to this region (nucleotide 684-698) were added to medium of cultured H1 delta 9 cell. HDAg and HDV cDNA were detected by ELISA and Dot blot hybridization. RESULTS: Twenty fourth hour after 6 mumol/L ASODN and S-ASODN were added, both the secreting amount of HDAg in the supernatant and HDV RNA were inhibited. The inhibiting rates were 76.14% and 84.50% respectively. When the concentration of S-ASODN was 2, 4, 6 mumol/L, the inhibiting rate showed the feature of dosage dependence. Inhibitory effect of ASODN and S-ASODN were similar in the same dosage. CONCLUSION: The results suggest that ASODN and S-ASODN can availably inhibit replication and expression of HDV in H1 delta 9 cell. PMID- 10366976 TI - [Effect of mutation (T1762A1764) on hepatitis B virus core promoter activity]. AB - OBJECTIVE: With the knowledge that the substituted mutation of T1762A1764 in hepatitis B virus (HBV) core promoter (CP) region was the most common variation during chronic HBV infection, we proceed to study the role that the mutation T1762A1764 plays in effecting the core promoter. METHODS: Seven patients with fulminant hepatitis and one asymptomatic HBV carrier were investigated for screening T1762A1764 mutation by cloning and PCR products direct sequencing. PCR products containing HBV precore/core gene regulatory sequences were directly cloned into chloramphenicol acetyltransferase (CAT) expressing plasmid. CAT assay was done to compare CAT expressing level after transfecting into HepG2 cell line and transient expression. RESULTS: Results reveal that T1762A1764 mutation was the main factor that represses CAT expression in vitro. CONCLUSION: Among the several other variations within hepatitis B virus' core promoter region, T1762A1764 mutation has mainly affected the down-regulating activity of core promoter. PMID- 10366977 TI - [The clinical and pathology study of HBV precore mutant infection in chronic HBV patients]. AB - OBJECTIVE: To elucidate the clinical feature, pathological change and IFN therapy of HBV pre-c mutant(1,896 G-->A point mutation). METHODS: Mutation specific PCR was employed for detecting precore mutation from 108 serum and/or liver samples of patients with chronic hepatitis B infection. Liver damage was evaluated with Knodell's histology activity index(HAI) in 46 patients. RESULTS: HBV precore mutant existed popularly in different e systems status, but the detective rate of HBV precore mutant or mutant prevailing over wild type in HBeAb+ group(14.29%, 46.23%) was significantly higher than that in HBeAg+(0%, 6.45%)group. Mutant infection was associated with the severity of liver diseases, the detective rate in severe chronic hepatitis group was significantly higher(13.64%, 40.91%) than that in chronic asympotamatic carriers groupe(0%, 15.62%). 5 precore mutant positive patients with IFN treatment showed either response or nonresponse to IFN therapy. There was no significant difference of liver pathological change between mutant type and wild type infection in patients with mild chronic hepatitis. CONCLUSION: HBV precore mutant infection existed commonly in patients with chronic hepatitis B, despite of the e system status, though it was more prevalent in patients with HBeAb positive. Further study is needed in the pathogenicity and the treatment of HBV precore mutation. PMID- 10366979 TI - [Helicobacter pylori and congestive gastropathy]. AB - OBJECTIVE: This study evaluates the prevalence and significance of Helicobacter pylori(H. pylori) infection in patients with portal hypertension. METHODS: 177 patients were selected. Among whom, 135 patients with portal hypertension and 42 noncirrhotic patients with nonulcerative dyspepsia as a control group. In all patients diagnostic upper endoscopy was performed and gastric biopsies were taken for histological examination and diagnosis of H. pylori infection. RESULTS: Of the portal hypertensive patients, 63 patients had congestive gastropathy, 17(27%) of whom were positive for H. pylori infection and 72 patients did not have gastropathy, 18(25%) of whom were positive for H. pylori infection. In the control group 23 of 42(55%) were positive for H. pylori infection. H. pylori was found less frequently in congestive gastropathy patients than in the control group. We also found that the presence and severity of congestive gastropathy is independent of the H. pylori status. CONCLUSION: The role of H. pylori infection in the pathogenesis of congestive gastropathy seems to be unlikely and we suggest that there is no need for its routine eradication in cirrhotic patients. PMID- 10366978 TI - [Relationship between the expression of transforming growth factor beta type I receptor (T beta R I) and prognosis of hepatocellular carcinoma (HCC)]. AB - OBJECTIVE: To study the expression of T beta R I in HCC tissues and its clinical significance. METHODS: Expressions of T beta R I in HCC tissues (HT, n = 30) and surrounding liver tissues(HST, n = 30) as well as four normal control liver tissues (CIT, n = 4) were determined by radioligand binding assay with 125I TGF beta 1 as the ligand. Relationships between some clinicopathological parameters of HCC and the relative expression rates of T beta R I in HCC were studied. RESULTS: Compared with that in HST, the expression of T beta R I HT was significantly lower than in HST and CIT (P < 0.01). The expression of T beta R I in HST is similar to that in CIT. T beta R I relative expression rates in the group of no capsule or capsule invaded and in the group of tumor embolus were markedly decreased (P < 0.05). No close relationships were found between the T beta R I relative expression rate and age, sex, HBV infection, liver cirrhosis, AFP concentration, tumor diameter, tumor number, tumor differentiation as well as tumor embolus. CONCLUSIONS: The low expression of T beta R I in HT may be one of the key event which promote the HCC malignant growth by protecting them against the growth inhibition effect of active TGF beta. The T beta R I relative expression rate may be related to the prognosis of HCC. PMID- 10366980 TI - [Serological character of hepatitis E virus mutants]. AB - OBJECTIVE: 64 sera with serologically negative nonA-E were tested for HEV RNA and their serological characteristics were studied. METHODS: HEV RNA was tested by a reverse transcriptase PCR(RT-PCR) and the positive PCR products were cloned and sequenced. Of 15 patients positive for HEV RNA, 6 were followed up and tested for anti-HEV. RESULTS: 15 of them(23.4%) were HEV RNA positive. 9 of the 15 HEV sequences have 95% homology and the remaining 6 have only 80% homology as compared with the Chinese HEV strain at the nucleotide level. Of 15 patients positive for HEV RNA, 6 were followed up. 3 of 4 patients infected with typical HEV had anti-HEV seroconversion 1 month after infection and 2 patients infected with HEV mutants were persistently negative for anti-HEV 1 or 3 months after infection. CONCLUSION: The Commercial anti-HEV EIA Kit can not detect HEV mutants. PMID- 10366981 TI - [Comparison of hypervariable region gene of hepatitis C virus between an individual infected persistently and an individual recovered from infection]. AB - OBJECTIVE: To explore the law of recovery from HCV infection. METHODS: Amplifiying, sequenceing and analyzing the gene of hypervariable region from an individual infected persistently and an individual recovered from infection. RESULTS: One base insertion (two clones, 10%) in the person recovered other than that infected persistently, but there is no difference between them in percentage of mumation, diversity of quasispecies and genetic distance among clones. CONCLUSION: The range of genetic variation at one time point did not seem to have obvious relationship with the HCV persistent infection or recovery from HCV infection. This study gave basic information for searching the relation of genetic variation and clinical status, and designing HCV vaccine. PMID- 10366982 TI - [Genotyping of HCV isolates from different populations in Shanghai by using second generation line probe assay]. AB - OBJECTIVE: To investigate the distribution and frequency of various genotypes in different populations in Shanghai. METHODS: 109 HCV isolates by using RT-nested PCR including 81 from patients with hepatitis C (PHCs), 7 from blood donors (BDs) and 21 from intravenous drug abusers (IVDAs) were genotyped by using recently improved second generation line probe assay (INNO-LIPA HCV II innogenetics N.V.Belgium). RESULTS: Among 81 HCV isolates from PHCs, there were 71 (87.6%) genotype 1b, 4(4.9%)2a, 2(2.5%)3b, 1(1.2%)6a, 2(2.5%) mixed genotypes (1 for 2a or 2c + 2b + 1, 1 for 1b + 6a) and 1 (1.2%) undeterminable respectively, and among 7 from BDs, there were 5 1b and 2 genotype 2a. But among 21 from IVDAs, there were 8(38.1%) genotype 1a, 5(23.8%)1b, 2(9.5%)2a or 2c, 2(9.5%)3a, 1(4.8%) 3b and 3(14.3%) mixed genotypes (1 for 1a + 2a or 2c, 1 for 1b + 2a or 2c + 6a and 1 for 1a + 1b + 3b) respectively. CONCLUSION: (1) 7 HCV subtypes (1a, 1b, 2a, 2b, 3a, 3b and 6a) belonging to 4 HCV genotypes (1, 2, 3, 6) were present in different populations in Shanghai. Of them, 2b, 3a, 3b and 6a were first found. (2) HCV genotype 1b was most commonly found in PHCs and BDs. 1a and 3a were only found in IVDAs. (3) The fact that more HCV subtypes were found in IVDAs in this city. PMID- 10366984 TI - [Apoptosis in MT2 cells induced by HepG2.2.15 cells]. AB - OBJECTIVE: To obtain the apoptosis in MT2 cells induced by HepG2.2.15 cells expressing Fas Ligand. METHODS: A human T-lymphotropic virus type I infected cell line MT2 was cultured overnight, then 0.5 ml serum contained hepatitis B virus DNA(HBV DNA), E antigen and S antigen was added to it together with 1 ml of fresh RPMI 1640 medium and incubated for 20-24 hours at 37 degrees C. The cells were washed three times with RPMI 1640 medium without serum, followed by continued incubation. The cells were harvested and Fas antigen were determined by immunohistochemistry. The cells expressed Fas antigen was added to HepG2.2.15 cells expressing Fas Ligand together with fresh RPMI 1640 medium and incubated for 48-72 hours at 37 degrees C. The apoptosis in MT2 cells was detected by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL). RESULTS: The Fas antigen was observed in MT2 cells harvested at 8-12 days post infected, and the strong signals of apoptosis was observed in MT2 cells incubated together with HepG2.2.15 cells for 48-72 hours. CONCLUSION: MT2, a human T cell line infected with hepatitis B virus is able to express Fas antigen. The apoptosis in MT2 cells induced by HepG2.2.15 indicated that the T-lymphocyte in the quality of effective cell and the hepatocyte in the quality of target cell could be reversed in the pathogenesis of viral hepatitis. PMID- 10366983 TI - [Liver support effect of experimental extracorporeal bioartificial liver]. AB - OBJECTIVE: To establish an extracorporeal bioartificial liver support system (EBLSS) and study its support effect for fulminant hepatic failure (FHF). METHODS: The EBLSS consists of three parts: human liver cells (hepatocytes, nonparenchymal hepatocytes) isolated and cultured by extracorporeal two-stage perfusion and spheroidal aggregates methods respectively, hollow fiber bioreactor, and supplementary circulation unit, which was employed to support the FHF model dogs. RESULTS: Compared with the control dogs treated with EBLSS but without liver cells, the survival time of artificial liver support dogs was significantly prolonged, the changes of blood pressure, heart rate and ECG were slower, and the serum ammonia and lactate levels were significantly lower at the 3 and 5 hr. In addition, a good viability of human liver cells was noted after 5 hr experiment. CONCLUSION: This EBLSS has played the metabolic role of cultured heman hepatocytes, is capable of compensating the function of the liver, and might provide effective artificial liver support and therapy for patients with FHF or severe hepatitis. PMID- 10366985 TI - [Improvement in affinity of phage antibodies against HBsAg by light chain shuffling]. AB - OBJECTIVE: We had cloned an Fd gene of human antibody against HBsAg previously. The objective of this experiment is to improve the affinity phage antibody by chain-shuffling. METHODS: Human antibody light chain gene repertoire was generated by RT-PCR from human peripheral blood lymphocyte, and then phage antibody sublibrary was constructed by inserting the repertoire into the phagmid which contained the Fd gene. The matched gene of light chain was selected by biopaning. RESULTS: After three rounds of selection, eight clones with higher absorbance than that of original clone at 490 nm in ELISA were obtained. DNA sequencing showed three of the five VL genes were kappa type, and the other two were lambda type. CONCLUSION: The results indicated that the affinity of chain shuffled phage antibodies(phab) were improved. The specificity of the selected phab was also confirmed. PMID- 10366986 TI - [Inhibitory effect of IGF-II antisense RNA on malignant phenotype of hepatocellular carcinoma]. AB - OBJECTIVE: Inhibitory effect of insulin like growth factor II (IGF-II) antisense RNA on malignant phenotype of hepatic cancer cells was studied. METHODS: A 0.1 kb cDNA of human IGF-II was reversely inserted into a eukaryotic expression vector of pcDNA3 and an IGF-II antisense RNA expression vector of pIGF-II As was obtained. The pIGF-II As was then introduced into human hepatic cancer cell line SMMC-7721. The effect of IGF-II antisense RNA which was expressed by pIGF-II As on SMMC-7721 cell cycles progression was measured with FCM. RESULTS: The coloning formation in anchorage-independent assay of SMMC-7721 cells with pIGF-II As was significantly decreased by comparison with control groups of SMMC-7721 cells and SMMC-7721 cells transfected with pcDNA3 vector alone. FCM analysis showed that the S phase of cell cycles for SMMC-7721 cells with pIGF-II As was increased, but there were no obvious changes in the control groups of SMMC-7721 cells and SMMC 7721 cells with pcDNA3. CONCLUSION: The pIGF-II As of IGF-II antisense RNA acts as an inhibitor on carcinogenesis of the SMMC-7721 cells in vitro. PMID- 10366987 TI - [Inhibitory effect of glycyrrhizin on NF-kappa B binding activity in CCl4 plus ethanol induced liver cirrhosis in rats]. AB - OBJECTIVE: To investigate the effects of Potenlini on nuclear factor-kappa B (NF kappa B) binding activity in the livers of animals models with liver cirrhosis, and to delineate the molecular mechanism of the bioactivities of Potenlini. METHODS: Male SD rats were randomly allocated into a normal control group, a model control group, and a Potenlini group. Rats in the latter two groups were treated with CCl4 and Ethanol solution in order to induce chronic liver injury. Rats in Potenlini group were given Potenlini treatment at the same time. All rats were killed at the 9th week after CCl4 administration. Serum and liver specimens were collected, serum ALT activities and histological findings were assessed. Nuclear extracts from liver tissues were prepared and gel retardation assays were performed for the evaluation of NF-kappa B activity. RESULTS: (1) Serum ALT levels were significantly reduced in rats treated with Potenlini compared with those in rats of the model control group, which had dramatically increased ALT levels. (2) Histologically, liver steatosis and fibrosis were severe in the rats of the model group, but were significantly improved in rats of the Potenlini group. (3) NF-kappa B binding activity was markedly increased in the liver specimens taken from the rats of the model control group in comparison with the binding of normal livers, but the binding levels were nearly normal in the livers of the Potenlini group. CONCLUSION: Potenlini can inhibit the NF-kappa B binding activity in CCl4 and ethanol induced chronic liver injury, and that may partially be the mechanism by which Potenlini protects liver from hepatotoxin-induced liver injury and cirrhosis. PMID- 10366989 TI - [Glycodeoxycholic acid changes the membrane fluidity and superoxides of lipids in hepatocytes]. AB - OBJECTIVES: To explore the mechanisms of hepatocyte injury caused by glycodeoxycholic acid (GDCA) through studies on the roles of GDCA to membrane fluidity and superoxides of lipids of hepatocyte. METHODS: GDCA was used to treat hepatocytes of rat cultured in vitro and changes in cell membrane fluidity and malondialdehyde(MDA) were observed. RESULTS: In the presence of GDCA (final concentration of 250 mumol/L), the concentration of hepatocyte MDA of GDCA increased significantly (P < 0.001) and membrane fluidity decreased (P < 0.02) after culture of 1 or 4 hours. ALT was directly related to MDA and fluorescence polarization (P), r was 0.945 and 0.986, respectively. CONCLUSION: GDCA can induce the increase of MDA and decrease of fluidity of hepatocyte membrane, and finally cause hepatocyte injury. PMID- 10366988 TI - [Dynamic changes of nitric oxide and biochemistry in rats with intrahepatic cholestasis]. AB - OBJECTIVE: To investigate the correlation between nitric oxide and biochemical changes of intrahepatic cholestasis. METHODS: Studies were made on endogenous NO, biochemical changes of cholestasis, liver mitochondrial function and renal excretion in a rat model at different time points after ANIT administration. RESULTS: In acute reactive phase, serum bilirubin, ALT, AKP, and bile acids significantly increased, accompanied by enhanced plasma NO and renal excretion. Liver NO increased and mitochondrial SDH activity decreased for 2-6 days after ANIT. There was a high negative correlation between liver NO and mitochondrial SDH activity (r = -0.92). CONCLUSION: These results show that higher liver NO may be an important mediator of intrahepatic cholestasis. However, plasma NO increase may play a role in rapidly decreasing serum bilirubin and restoring bile acids, ALT, and AKP to control levels by mediating enhanced renal excretion. PMID- 10366990 TI - Is there an interaction between H2-antagonists and alcohol? AB - H2-antagonists are commonly prescribed drugs and alcohol use is widespread in the community. Any possible interaction may be important because of the frequent co administration of both drugs and the potential for unexpected impairment of pyschomotor function, in particular, driving skills. Hepatic ADH is the major site of alcohol metabolism. ADH is also found in the stomach, but it is uncertain whether gastric ADH is able to metabolise a significant amount of alcohol in vivo. Significant first-pass metabolism can be demonstrated at lower doses of alcohol, and if alcohol is given after meals. Varying degrees of extraction of alcohol from the portal circulation probably explains the data regarding first pass metabolism rather than gastric metabolism by gastric ADH. H2-receptor antagonists inhibit gastric ADH activity to a variable extent. If gastric metabolism of alcohol is negligible then this inhibition has no relevance. Given the uncertainty regarding a mechanism of interaction, only carefully conducted studies in controlled environments will answer the question. The large inter subject variability of alcohol absorption means that any study which seeks to determine the effect of an H2-receptor antagonist on ethanol metabolism must have sufficient numbers. A cross-over design, with each subject acting as his own control, is preferable to avoid ascribing an effect to treatment rather than to chance. The alcohol dosing studies are reviewed and the results summarised according to dose of alcohol given. At a dose of 0.15 g/kg of alcohol, four commonly used H2-antagonists may cause a small increase in blood alcohol concentrations in certain conditions. This absolute increase is very small. The magnitude of effect is far less than the effect of taking a meal before alcohol. At doses of 0.3 g/kg and above the majority of evidence favours no interaction between H2-antagonists and alcohol. There is no interaction at doses that would be expected to impair psychomotor skills (above 25 mg/dl). There remains a question regarding the cumulative effect of repeated small doses of alcohol and further studies are required. The relationship between ethanol absorption and gastric emptying raises the possibility that the effects of H2-receptor antagonists observed at very low doses of alcohol may be due to the acceleration of gastric emptying by these drugs. This is an attractive hypothesis that explains many aspects of the debate, but studies of the effect of H2-antagonists on gastric emptying have been conflicting. PMID- 10366991 TI - Epidemiology, outcomes research, and drug interactions. AB - Drug interactions are one of several outcomes (typically adverse) associated with drug therapy. The bulk of the clinical literature that has been published about drug interactions has focused either on the mechanisms, pharmacokinetic studies, or cases observed by clinicians. However, when it comes to the true risk of a drug interaction, or the probability of such an interaction occurring in a given population, a dearth of information is available. This area is opportune for the application of epidemiological and outcomes-based studies. PMID- 10366992 TI - Absence of interaction between tiagabine, a new antiepileptic drug, and the benzodiazepine triazolam. AB - In a randomised, double blind, placebo-controlled, four-period cross-over study in 12 healthy volunteers, the potential pharmacodynamic and pharmacokinetic interactions between the new antiepileptic drug, tiagabine, and the benzodiazepine, triazolam, were investigated. A single dose of tiagabine HCl 10 mg did not enhance the sedative or cognitive effects of a single dose of the benzodiazepine triazolam 0.125 mg, although the time-course of the effects was prolonged. Furthermore, tiagabine did not produce any statistically significant effects on the pharmacokinetics of triazolam. Similarly, the pharmacokinetics of tiagabine were not modified by triazolam. Tiagabine was well tolerated when administered alone or with triazolam. PMID- 10366993 TI - Pharmacokinetics and hypotensive effect of diltiazem in rabbits after a single intravenous administration: effect of phenobarbital. AB - Metabolism of the widely used calcium antagonist diltiazem (DTZ) is an important contributing factor to its therapeutic effects. In order to study the effects of CYP3A induction on the pharmacokinetics and haemodynamic effect of DTZ, it was administered as a single 5 mg/kg dose i.v. to two groups of New Zealand white rabbits (n = 6 in each group). Prior to the injection, one of the groups received phenobarbital 20 mg/kg s.c. two times a day for 3 days to ensure CYP3A induction, and the other received normal saline. A third group of animals (n = 6) received neither phenobarbital nor DTZ, and served as the control. Blood samples, systolic and diastolic blood pressure (SBP and DBP), and heart rate (HR) recordings were obtained from each rabbit up to 7 h, and urine samples for 48 h post-dose. Plasma concentrations of DTZ and its metabolites were determined by HPLC. The results showed that phenobarbital increased the Cl and Vdss of DTZ from 24 +/- 14 to 51 +/- 4.9 ml/min/kg and from 1.9 +/- 1.2 to 3.8 +/- 0.7 l/kg, respectively (p < 0.05). It also decreased the plasma concentrations of DTZ and all the measured metabolites in this study. Both phenobarbital and DTZ decreased SBP and DBP significantly without affecting the HR. PMID- 10366994 TI - Effect of clopidogrel on naproxen-induced gastrointestinal blood loss in healthy volunteers. AB - The effect of clopidogrel, a potent inhibitor of platelet aggregation, on naproxen-induced faecal blood loss was investigated in 30 healthy volunteers in a randomized, double-blind, placebo-controlled, two parallel treatment groups study. All subjects first received naproxen 250 mg b.i.d. during 7 days, after which they were randomly allocated to additionally receive either clopidogrel 75 mg once daily (n = 15) or matching placebo (n = 15) for 11 days. Faecal blood loss was measured by the 51Cr-labelled erythrocyte method during the last four days of each of the four study periods, i.e. baseline, treatment with naproxen alone, combined treatment and wash-out. Mean daily faecal blood loss was below 0.5 ml/day during the baseline period in both treatment groups and increased during treatment with naproxen alone to (mean +/- SD) 1.14 +/- 0.58 ml/day in the naproxen + placebo group and to 1.93 +/- 1.51 ml/day in the naproxen + clopidogrel group. Addition of clopidogrel to naproxen treatment was associated with an increase of the mean daily blood loss to 6.83 +/- 9.32 ml/day, which was statistically significantly higher than 1.75 +/- 1.40 ml/day observed during treatment with naproxen + placebo. During the wash-out period, mean daily blood loss decreased to 0.98 +/- 0.51 ml/day in the naproxen + placebo group and to 1.07 +/- 0.46 ml/day in the naproxen + clopidogrel group. Based on these results, it can be concluded that clopidogrel increases naproxen-induced gastrointestinal blood loss in healthy volunteers. Caution should therefore be called for when these drugs are coadministered. PMID- 10366995 TI - Validation and use of the CALUX-bioassay for the determination of dioxins and PCBs in bovine milk. AB - There is a strong need for the development of relatively cheap and rapid bioassays for the determination of dioxins and related compounds in food. A newly developed CALUX (Chemical-Activated LUciferase gene eXpression) bioassay was tested for its possible use to determine low levels of dioxins in bovine milk. Data show that this mammalian cell-based test is very sensitive for 2,3,7,8 substituted dioxins and related PCBs, thereby reflecting the relative potencies of these compounds in comparison to TCDD (TEF-values). The limit of detection was about 50 fg of TCDD. Furthermore, the response obtained with a mixture of dioxins was additive, in accordance with the TEF-principle. Milk fat was isolated by centrifugation followed by clean-up of the fat with n-pentane, removal of the fat on a 33% H2SO4 silica column, and determination of Ah receptor agonist activity with the CALUX-bioassay. An equivalent of 67 mg fat was tested per experimental unit, resulting in a limit of quantification around 1 pg i-TEQ/g fat. To investigate the performance of the method, butter fat was cleaned and spiked with a mixture of 17 different 2,3,7,8-substituted PCDD and PCDF congeners at 1, 3, 6, 9, 12 and 15 pg TEQ/g fat, as confirmed by GC/MS. In this concentration range, the method showed a recovery of TEQs around 67% (58-87%). The reproducibility, determined in three independent series showed a CV varying between 4% and 54%, with the exception of the sample spiked at 1 pg i-TEQ (CV 97%). The repeatability determined with the sample spiked at 6 pg i-TEQ/g showed a CV of 10%. Testing of 22 bovine milk samples, taken at different sites in The Netherlands, in the CALUX assay showed combined dioxin and dioxin-like PCB levels equivalent to 1.6 pg TCDD/g fat (range 0.2-4.6). GC/MS analysis of these samples revealed an average level of 1.7 pg i-TEQ/g fat, varying between 0.5 and 4.7 pg i-TEQ/g fat. All five samples showing a GC/MS determined dioxin content of more than 2 pg i-TEQ/g fat gave a response in the CALUX-assay corresponding to more than 2 pg TCDD/g fat. These data clearly show that the CALUX-bioassay is a promising method for the rapid and low cost screening of dioxins in bovine milk. PMID- 10366996 TI - Total mercury in muscle of benthic and pelagic fish from the South Adriatic Sea (Italy). AB - Total mercury concentrations were measured from the muscle of different kinds of fish: yellow gurnard (Trigla lucerna), red gurnard (Aspitrigla cuculus) red fish (Helicolenus dactylopterus), skate spp. (Raje spp.), goldline (Sarpa salpa), atlantic bonito (Sarda sarda), mackerel (Scomber scombrus), chub mackerel (Scomber japonicus) caught in the South Adriatic Sea (south Italy) in the period June/August 1995. The highest total mercury levels were found in the benthic marine organisms and particularly in skates (Raje spp.) whose values ranged from 0.05 to 2.65 mg/kg wet wt with a mean value of 1.02 mg/kg wet wt. As for pelagic species, the highest mean levels were observed in Atlantic bonito (0.34 mg/kg wet wt), while in goldline the mean content of total mercury was the lowest (0.07 mg/kg wet wt). According to the rules in force (Official Journal of the European Communities 1994) 53% of skate and Atlantic bonito samples showed concentrations exceeding the peak value of 1 mg/kg, while for the other species, only 28% of samples exceeded the peak value fixed at 0.5 mg/kg. Correlations between total mercury concentration and specimen weight were evident in all examined species except for goldline and skates. PMID- 10366997 TI - Toxic and essential elements in fish from Nordic waters, with the results seen from the perspective of analytical quality assurance. AB - Fish from fresh, brackish and marine waters were analysed for their levels of Pb, Cd, Cr, Ni, Co, Zn, Cu, Mn, As and Se in muscle tissue. Various quality control procedures were implemented during the survey and the results were compared with those from several other surveys, most of which were carried out during the 1990s. It was noted that several elements varied widely both within and between studies. A systematic examination of a large number of surveys and ranking of their quality level indicated that a lack of quality control procedures often results in high or varying results. This may have serious consequences when results are used as a basis for for example, legislation or intake recommendations. PMID- 10366998 TI - Concentrations of fluoride in wines from the Canary Islands. AB - Potentiometry using an ion-selective electrode has widely been used for determining fluoride because of its simplicity and rapidity. The concentration of fluoride was determined (Gran's method) in 70 wines from the main wine-producing regions of the Canary Islands. The mean concentration of fluoride in wines from a region with a high concentration of fluoride in drinking waters was significantly (p < 0.05) higher than those mean concentrations obtained in the remaining wines. Non-important differences were found among the types of wine analysed. However, the fluoride concentrations of all the Canarian wines analysed here did not present a risk for the public health. PMID- 10366999 TI - Determination of borates in caviare by ion-exclusion chromatography. AB - A specific and rapid analytical procedure for the determination of borates in caviare was developed using ion-exclusion chromatography. Products require minimal pre-treatment, are simply extracted with eluent and filtered prior to chromatography. Isolation of the analyte as a 2:1 sorbitol-borate complex was accomplished using an anion exchange column with an eluent consisting of 2 mM heptafluorobutyric acid + 50 mM sorbitol with conductivity detection [corrected]. Among five sugar alcohols examined for complexation, sorbitol was found to provide optimum resolution and a 3-4 times enhancement of response over borate anion alone. The response of the complex was linear over at least a 100 fold range in concentration (0.1-10 micrograms boron/ml) with a correlation coefficient of 0.9997. Using a boric acid standard solution, the limits of detection and quantitation were 0.065 microgram/ml and 0.216 microgram/ml, respectively, calculated as boron. Replicate analyses (n = 5) for samples having commercially added borate (equivalent to 300-1000 micrograms/g boron) gave RSD values of 1.18-2.03%. Recoveries of borate from fortified samples having commercially added borate present varied from 94.5 to 101.1% while untreated samples exhibited recoveries of 78.5-108.0% over a concentration range of 23-1039 micrograms/g, calculated as boron. PMID- 10367000 TI - Quality of mechanically deboned chicken meat frankfurter incorporated with chicken skin. AB - Four formulations were processed into frankfurters with different ratios of mechanically deboned chicken meat (MDCM) and cooked chicken skin (CCS) i.e. 80/0, 70/10, 60/20 and 50/30. The products were evaluated for proximate composition, cholesterol content, colour; 'L' value (lightness) and 'a' value (redness), percentage of cooking loss, physical measurements (shearforce-kgf and folding test), thiobarbituric acid value (TBA) and taste panel evaluation. The increment of CCS in the frankfurters increased the contents of moisture, ash, protein, fat, cholesterol, the lightness ('L' value) and redness ('a' value). After 3 months of frozen storage, the increment continued except for the moisture contents for formulations with 20 and 30% CCS. The lipid oxidation (TBA value) and cooking loss were lowered in formulations with CCS. After 3 months of frozen storage, TBA value decreased, while the cooking loss increased for all the formulations. The addition of CCS increased hardness of the frankfurters but affected folding ability, with formulation with 10% CCS scoring better grade. Sensory evaluation was carried out using 30 untrained panelists to evaluate aroma, colour, appearance, hardness, juiciness, chicken taste, oily taste, rancid taste and overall acceptance of the products. The addition of CCS in the frankfurters at 10 and 20% resulted in products with taste and texture that were acceptable after 3 months of frozen storage. PMID- 10367001 TI - Replacement of animal fat with fractionated and partially hydrogenated palm oil in beef burgers. AB - Four formulations of burgers, prepared with 65% lean meat and 15% fat consisting of RBD palm stearin (PS), Socfat 4000P and Socfat 4100P and beef fat (BF) as control were evaluated for solid fat content (SFC), slip melting point (SMP), cooking loss, proximate analysis (moisture, fat and protein), colour, i.e. lightness ('L'), redness ('a') and yellowness ('b'), free fatty acid (FFA), iodine value (IV), thiobarbituric acid (TBA) and texture profile analysis (TPA). Sensory evaluation was carried out for texture, juiciness, aroma, oiliness and overall acceptance. SFC and SMP for raw and cooked SF4000P beef burgers were closest to BF control burgers, falling into the range of 35-40 degrees C. Cooking loss was highest for PS burgers, there were no significant differences (P > 0.05) amongst BF, SF4000P and SF4100P burgers. Proximate analysis on raw burgers showed SF4000P to contain high fat and lowest moisture contents. Objective textural measurements using texture profile analysis (TPA) for all cooked burgers showed no significant differences (P > 0.05) for springiness and cohesiveness. Variation of values among the formulations for hardness, gumminess and chewiness are explained by the differences of SFC for beef burgers with various types of fats. Raw and cooked PS burgers have the lightest 'L' values compared with other fat substituted burgers while BF, SF4000P and SF4100P indicated no significant differences (P > 0.05) for 'L', 'a' and 'b' values. Beef fat showed the highest amount of free fatty acids (FFA) compared to palm oil samples. For the iodine value (IV), SF4000P showed the highest value which means that it contained the highest level of unsaturated fatty acids followed by PS, BF and SF4100P successively. SF4000P had the highest TBA values followed successively by BF, PS and SF4100P. For sensory evaluation, PS burgers had the least oily taste. This may be due to its high cooking loss. Taste panelists could not differentiate burgers with substituted vegetable fats against the control burgers. PMID- 10367002 TI - Assessment of cyanogenic glucoside (cyanide) residues in Mbege: an opaque traditional Tanzanian beer. AB - Levels of cyanide in two varieties of malted millet, spent grain (Machicha) and opaque beer (Mbege) were determined. Protein content and amino acid composition of the malt, Mbege and Machicha were determined. Mbege was made in the laboratory using an improved method. The cyanide content of millet, malt, spent grain and Mbege were 40.6, 513.4, 18.9 and 8.1 ppm, respectively for the Moshi local millet variety. For Sumbawanga-2 millet variety the cyanide content was found to be 41.2, 489.2, 17.8 and 6.8 ppm for the millet, malt, spent grain and Mbege, respectively. The cyanide content increased linearly as the number of days of germination of the millet grain increased and the highest values of cyanide were attained on the third day of germination. Cyanogenic glucosides in the millet were enzymetically hydrolysed to respective cyanohydrins and volatile hydrogen cyanide due to low pH level of the Mbege which was 4. Malting of the millet increased the protein content by 5%. Lysine, the most limiting amino acid in millet, increased by 20%. It was concluded that the fermentation process of the millet malt into Mbege is efficient in reducing the levels of cyanogenic glucosides below levels considered toxic and therefore rendering the product safe. PMID- 10367003 TI - The effect of intermittent heating on some chemical parameters of refined oils used in Egypt. A public health nutrition concern. AB - As part of a public health campaign in Egypt, various chemical parameters of oil which are considered good indices in assessing the degree of thermal abuse, oxidation and overall quality (acid values, iodine values, peroxide values, etc.) were studied with respect to different frying oils. Ingestion of decomposition products formed as a results of thermal abuse and oxidation of frying oils are known to lead to a variety of symptoms and diseases (allergies, atherosclerosis, coronary heart disease). Results show that the oil most commonly used by street vendors in Egypt (blend of cotton seed and sunflower oil) is the least suitable for frying, while palm oil on the basis of the various chemical parameters studied, is the ideal choice. However, from the nutritional and public health stand point, the use of saturated oils is to be discouraged. Corn oil is therefore the next best choice from both the chemical and nutritional stand point, and is recommended for public use in a country in which deep-fried vegetable patties ('fallafel') forms the staple food item in the diet. PMID- 10367004 TI - Lipid content and fatty acid composition in foods commonly consumed by nursing Congolese women: incidences on their essential fatty acid intakes and breast milk fatty acids. AB - The fat content and fatty acid (FA) composition of nearly 40 foods, currently consumed by 102 nursing Congolese mothers living in Brazzaville, were determined to assess their impact on mothers' essential fatty acid (EFA) intakes and breast milk FA. Data on mothers' milk FA and dietary habits which allowed food selection were recently published (Rocquelin et al., 1998). Most foods were locally produced. Food samples were collected at local markets, bleached if necessary to avoid microbial degradation, and stored at +4 degrees C or -20 degrees C. They were lyophilized upon their arrival in the laboratory before lipid analyses. FA composition of food lipids was determined by capillary gas chromatography. Staple diets included low-fat, high-carbohydrate foods (processed cassava roots, wheat bread) and high-polyunsaturated fatty acid (PUFA) foods: soybean oil (high in 18 : 2 n-6 and alpha-18 : 3 n-3), bushbutter (dacryodes edulis), peanuts, avocado (high in fat and 18 : 2 n-6), freshwater and salt-water fish (high in LC n-3 and/or n-6 PUFA), and leafy green vegetables (low in fat but very high in alpha 18 : 3 n-3). Their frequent consumption by nursing mothers provided enough EFA to meet requirements due to lactation. It also explains why mothers' breast milk was rich in C8-C14 saturated FA (26% of total FA) and in n-6, n-3 PUFA (respectively 15.0% and 2.4% of total FA) highly profitable for breastfed infants' development. From this point of view, dietary habits of Congolese mothers have to be sustained for they are more adequate than most Western-type diets. PMID- 10367005 TI - Trends and nutritional significance of mineral content in fresh white asparagus spears. AB - Variations in the copper, iron, zinc, manganese, calcium, magnesium, sodium, potassium and phosphorous content of fresh white asparagus (Asparagus officinalis, L.) as a function of the spear portion and the differences between varieties (Desto and Cipre's) and thicknesses (< 11 and > 14 mm) of asparagus were investigated. The mineral elements studied showed significant changes between the ten portions into which the spears were cut, with lower concentrations, except for sodium, occurring in the asparagus portions furthest away from the tip of the spear. These changes between portions signified a greater nutritional value of the apical spear area (6 cm from the asparagus tip) according to the higher percentages of mineral RDAs supplied in the apical portions. The varieties and thicknesses of white asparagus investigated showed this diminishing distribution of the mineral content throughout the spear and displayed significant differences for the majority of the mineral elements between varieties and thicknesses, although these differences were minimal and did not have any nutritional significance since the percentages of mineral RDAs supplied were similar. Nutrient density was well over 100% for all the elements, except for sodium, and these results indicated that fresh white asparagus, if consumed in sufficient quantities, contributes substantially to the intake of these mineral elements. PMID- 10367006 TI - Effect of processing on some properties of cowpea (Vigna unguiculata), seed, protein, starch, flour and akara. AB - Large brown eye Kano white cowpea (Vigna unguiculata) seeds were processed into three batches of flour by wetting, drying individually at 30, 80, and 120 degrees C, decorticating and dry milling. Starch was extracted from the cowpea seed and protein from the flour using water as solvent. The water-extractable proteins were purified by dialysis and analysed by electrophoresis. The cowpea flour was used to produce akara balls (fried paste). The microstructure of the cowpea cotyledon, flour, starch and akara crumb were examined with a scanning electron microscope (SEM). Electrophoretic separation revealed that some of the protein fractions from the sample extracted from 30 degrees C dried cowpea were absent in the sample extracted from the 80 degrees C and 120 degrees C dried cowpeas or their quantities had decreased. In the SEM study, no difference was observed in the microstructure of the three flour samples except in the size and shape of the starch granules and particles of protein and cell wall material. The starch granules from the cowpea dried at 120 degrees C had surface defects. Cavities occurred in the cotyledons of the 80 and 120 degrees C dried cowpea seeds, some starch granules, protein matrix and sometimes the entire cell contents were lost from the cell. The protein sheet in the akara crumb became thicker as temperature increased to 80 and 120 degrees C. PMID- 10367008 TI - The role of prophylactic iron supplementation in pregnancy. AB - The prevalence, causes and role of iron prophylaxis in pregnant women was studied. All women delivered at the National University Hospital, Singapore in 1993 had their haemoglobin estimated. If it was less than 11 g/dl, blood was taken for serum iron, ferritin, transferrin, red cell zinc protoporphyrin, serum folate, vitamin B12 and thalassemia screen to establish cause of anaemia. Data was also collected with regards to their antenatal progress and iron prophylaxis. Logistic regression, Chi-square test, Fischer's exact test and Mantel-Haenszel tests were also used to assess the relationships between categorical variables. The prevalence of anaemia at first antenatal visit was 20.6% while the prevalence of anaemia at delivery was 15.3%. The commonest cause of the anaemia in pregnancy was due to iron deficiency (81.3%). In the non-anaemic group, 90.7% were on prophylactic iron supplements compared to 50.6% in the anaemic group (P < 0.001). Of the 752 women found to be anaemic at booking, 591 received prophylactic iron supplements while 161 women did not. A total of 166 (28.1%) of those with iron supplements were anaemic at delivery, whereas 140 (87.0%) of those who did not receive prophylactic iron remained anaemic at delivery (P < 0.001). Of the 2516 non-anaemic women who received prophylactic iron, 118 (4.7%) developed anaemia at delivery while 133 (34.1%) out of the 390 women who did not receive prophylactic iron were anaemic at delivery (P < 0.001). Multivariate logistic regression analysis revealed the odds of anaemia for a woman not on iron therapy was about 11 times that of her counterpart on prophylactic iron therapy (95% CI 8.76 to 14.13). A 55% reduction in odds of anaemia was estimated per 1 gm% increase in haemoglobin at booking. Prophylactic antenatal iron supplements not only prevent a fall but also improved haemoglobin levels during pregnancy. Those who were not on any iron supplements were 11 times more likely to develop anaemia in the present pregnancy. PMID- 10367007 TI - Effect of food processing on iron availability of African pearl millet weaning foods. AB - The effects of different cereal precooking process (roasting and extrusion cooking) on iron availability and protein digestibility of four African weaning foods were investigated using in vitro methods. In two weaning foods based on pearl millet, cowpea and peanut, the cereal was extruded (A) or roasted (B). In two other weaning foods having a similar composition, a low proportion of milk powder was added and the cereal extruded (C) or roasted (D). The mean +/- SD iron values (mg/100 g) were: A, 5.56 +/- 0.18; B, 9.12 +/- 0.93; C, 5.89 +/- 0.23; D, 9.04 +/- 0.85. When the pearl millet was roasted, the iron content was higher than in the extruded weaning foods (P < 0.01). However, the percent of available iron of the roasted weaning foods was very low (B, 1.64 +/- 0.01; D, 0.91 +/- 0.02). The iron availability of the extruded weaning foods, A and C, was 3.5 times and 6.5 times higher than the corresponding roasted weaning foods, B and D, respectively. This represented 332.4 +/- 4.4 and 375.1 +/- 5.8 micrograms of available iron/100 g for A and C, respectively versus 149.5 +/- 0.9 and 82.2 +/- 1.8 micrograms of available iron/100 g for B and D, respectively. No significant differences in polyphenol contents were found according to the precooking process of the cereal. The extruded weaning foods showed a higher protein digestibility of approximately 10% than the roasted ones (P < 0.05). A positive correlation was found between in vitro iron availability and protein digestibility (r = 0.976, P < 0.02). Despite a high content of iron, the iron availability of roasted pearl millet weaning foods was quite low. Extrusion cooking of the cereal improved the protein digestibility and iron availability of pearl millet weaning foods; however, the amount of available iron remained insufficient to meet the iron requirements of infants whatever the cereal processing. PMID- 10367009 TI - Nutrient intake of rural pregnant women of Haryana state, northern India: relationship between income and education. AB - The daily nutrient intake of 90 pregnant women from farming and non-farming communities in six rural villages of Haryana State, Northern India have been determined. As a result of questionnaires and interviews, nutrient intake for 3 consecutive days were calculated. Mean daily intakes of farming and non-farming pregnant women examined in this study were lower for energy, calcium and iron than the recently prescribed Indian recommended dietary allowances (RDAs). Protein intake of non-farming women was significantly lower and that of farming women was almost similar to RDA. Intake of fat by pregnant women was double the RDA. The mean daily intakes of thiamine, riboflavin and niacin by women of both the communities were found to be adequate. The diets of pregnant women could meet half the requirement of folic acid and even less than half for ascorbic acid. Income of pregnant women did not show any influence on nutrient intakes but educational level of women certainly reflected differences in vitamin intakes. PMID- 10367010 TI - Breakfast and mental health. AB - The objective of the present investigation was to study the relationship between breakfast consumption and subjective reports of mental health and health-related behaviours in a general population sample (126 subjects aged between 20 and 79 years). Individuals who consumed a cereal breakfast each day were less depressed, less emotionally distressed and had lower levels of perceived stress than those who did not eat breakfast each day. Those who consumed breakfast had a healthier lifestyle than the others in that they were less likely to be smokers, drank less alcohol and had a healthier diet. However, the relationship between cereal breakfast consumption and mental health did not reflect these differences in the smoking, alcohol consumption and diet. In conclusion, there is an association between breakfast consumption and well-being which cannot entirely be accounted for by differences in other aspects of diet or smoking and alcohol consumption. Further intervention studies are now needed to establish whether causal relationships and mechanisms underlie the associations seen in this study. PMID- 10367011 TI - Altered mechanics of cartilage with osteoarthritis: human osteoarthritis and an experimental model of joint degeneration. AB - OBJECTIVE: Studies of cartilage mechanics seek to determine the fundamental relationships between mechanical behavior and the composition and structure of healthy cartilage and to determine mechanisms for changes associated with degeneration. METHOD: The mechanics of normal and osteoarthritic (OA) human articular cartilage are reviewed. Studies of the initiation and pathogenesis of cartilage degeneration in the anterior cruciate ligament transection (ACLT) model of joint instability are also presented. RESULTS: In human cartilage with OA, tensile, compressive and shear behaviors are dramatically altered. These changes present as decreases in the modulus or stiffness of OA cartilage in tension, compression and shear loading, and increases in the propensity to swell as compared to healthy cartilage. In the ACL transection model of OA, similar changes in the mechanics of cartilage have been observed. In addition, changes in structure, composition, and as metabolism consistent with human OA have been found. Deterioration of the collagen-proteoglycan solid network, which appears to be focused at the articular surface, has been the earliest cartilage changes in the model. It remains to be determined if the initial disruption of the cartilage surface is a direct result of mechanical forces or a product of altered chondrocyte activity. CONCLUSIONS: These data and continued research using experimental models of OA provide a basis for our understanding of the pathogenesis and the time course of events in OA and will lead to the development of better procedures for disease intervention and treatment. PMID- 10367012 TI - Articular cartilage repair: are the intrinsic biological constraints undermining this process insuperable? AB - This article reviews the experimental and clinical strategies currently in use or under development for the treatment of articular cartilage lesions. The vast majority of protocols under investigation pertain to the treatment of full thickness defects (i.e., those which penetrate the subchondral bone and trabecular-bone spaces) rather than partial-thickness ones (i.e., those which are confined to the substance of articular cartilage tissue itself). This bias probably reflects the circumstance that partial-thickness defects do not heal spontaneously whereas full-thickness ones below a critical size do, albeit transiently. And it is, of course, a seemingly easier task to manipulate a process which is readily set in train than it is to overcome an induction-problem which Nature herself has not solved. Indeed, the reasons for this inert state of partial-thickness defects have only recently been elucidated, and these are briefly discussed. However, the main body of this review deals with the various transplantation concepts implemented for the repair of full-thickness defects. These fall into two broad categories: tissue-based (entailing the grafting of perichondrial, periosteal, cartilage or bone-cartilage material) and cell-based (utilizing chondroblasts, chondrocytes, periochondrial cells or mesenchymal stem cells). Cell-based systems are further subdivided according to whether cells are transplanted within a matrix (biodegradable, non-biodegradable or synthetic) or free in suspension. Thus far, the application of cell suspensions has always been combined with the grafting of a periosteal flap. The strengths and weaknesses of each concept are discussed. PMID- 10367013 TI - Biomechanics of integrative cartilage repair. AB - Cartilage repair is required in a number of orthopaedic conditions and rheumatic diseases. From a macroscopic viewpoint, the complete repair of an articular cartilage defect requires integration of opposing cartilage surfaces or the integration of repair tissue with the surrounding host cartilage. However, integrative cartilage repair does not occur readily or predictably in vivo. Consideration of the 'integrative cartilage repair process', at least in the relatively early stages, as the formation of a adhesive suggests several biomechanical approaches for characterizing the properties of the repair tissue. Both strength of materials and fracture mechanics approaches for characterizing adhesives have recently been applied to the study of integrative cartilage repair. Experimental configurations, such as the single-lap adhesive test, have been adapted to determine the strength of the biological repair that occurs between sections of bovine cartilage during explant culture, as well as the strength of adhesive materials that are applied to opposing cartilage surfaces. A variety of fracture mechanics test procedures, such as the (modified) single edge notch, 'T' peel, dynamic shear, and trouser tear tests, have been used to assess Mode I, II, and III fracture toughness values of normal articular cartilage and, in some cases, cartilaginous tissue undergoing integrative repair. The relationships between adhesive biomechanical properties and underlying cellular and molecular processes during integrative cartilage repair remain to be elucidated. The determination of such relationships may allow the design of tissue engineering procedures to stimulate integrative cartilage repair. PMID- 10367014 TI - The extracellular matrix, interstitial fluid and ions as a mechanical signal transducer in articular cartilage. AB - OBJECTIVE: (1) Provide an overview of the biomechanical factors that are required to analyze and interpret biological data from explant experiments; (2) Present a description of some of the mechano-electrochemical events which occur in cartilage explants during loading. DESIGN: A thorough and provocative discussion on the effects of loading on articular cartilage will be presented. Five simplest loading cases are considered: hydrostatic pressure, osmotic pressure, permeation (pressure loading), confined compression and unconfined compression. Details of how such surface loadings are converted or transduced by the extracellular matrix (ECM) to pressure, fluid, solute and ion flows, deformation and electrical fields are discussed. RESULTS: Similarities and differences in these quantities for the five types of loading are specifically noted. For example, it is noted that there is no practical mechanical loading condition that can be achieved in the laboratory to produce effects that are equal to the effects of osmotic pressure loading within the ECM. Some counter-intuitive effects from these loadings are also described. Further, the significance of flow-induced compression of the ECM is emphasized, since this frictional drag effect is likely to be one of the major effects of fluid flow through the porous-permeable ECM. Streaming potentials arising from the flow of ions past the fixed charges of the ECM are discussed in relation to the flow-induced compaction effect as well. CONCLUSION: Understanding the differences among these explant loading cases is important; it will help to provide greater insights to the mechano-electrochemical events which mediate metabolic responses of chondrocytes in explant loading experiments. PMID- 10367015 TI - The deformation behavior and mechanical properties of chondrocytes in articular cartilage. AB - INTRODUCTION: Chondrocytes in articular cartilage utilize mechanical signals to regulate their metabolic activity. A fundamental step in determining the role of various biophysical factors in this process is to characterize the local mechanical environment of the chondrocyte under physiological loading. METHODS: A combined experimental and theoretical approach was used to quantify the in-situ mechanical environment of the chondrocyte. The mechanical properties of enzymatically-isolated chondrocytes and their pericellular matrix (PCM) were determined using micropipette aspiration. The values were used in a finite element model of the chondron (the chondrocyte and its PCM) within articular cartilage to predict the stress-strain and fluid flow microenvironment of the cell. The theoretical predictions were validated using three-dimensional confocal microscopy of chondrocyte deformation in situ. RESULTS: Chondrocytes were found to behave as a viscoelastic solid material with a Young's modulus of approximately 0.6 kPa. The elastic modulus of the PCM was significantly higher than that of the chondrocyte, but several orders of magnitude lower than that of the extracellular matrix. Theoretical modeling of cell-matrix interactions suggests the mechanical environment of the chondrocyte is highly non-uniform and is dependent on the viscoelastic properties of the PCM. Excellent agreement was observed between the theoretical predictions and the direct measurements of chondrocyte deformation, but only if the model incorporated the PCM. CONCLUSIONS: These findings imply that the PCM plays a functional biomechanical role in articular cartilage, and alterations in PCM properties with aging or disease will significantly affect the biophysical environment of the chondrocyte. PMID- 10367016 TI - Intermittent sub-ambient interstitial hydrostatic pressure as a potential mechanical stimulator for chondrocyte metabolism. AB - OBJECTIVE: Experimental findings have suggested that the metabolic activities of articular cartilage can be influenced by mechanical stimuli. Our mathematical analysis predicted that cyclic compressive loading may create periods of intermittent sub-ambient hydrostatic pressure within the cartilage extracellular matrix. Based on this mathematical analysis, the present study was aimed to investigate whether the intermittent sub-ambient hydrostatic pressure, created in the cartilage extracellular matrix during cyclic compression, has a stimulative effect on the biosynthesis of chondrocytes. METHOD: In order to test this hypothesis, the present study developed a custom-designed sub-ambient pressure generator to subject a monolayer culture of chondrocytes to an intermittent sub ambient pressure. Using this pressure generator, the monolayer chondrocyte culture system was analyzed for 35S-sulfate and 3H-proline incorporation rates for biosynthesis of proteoglycan and collagenous/noncollagenous protein molecules, respectively. Northern analyses for aggrecan and type II collagen mRNAs were also performed. RESULTS: It was found that the intermittent sub ambient pressure produced a 40% increase in proteoglycan and a 17% increase in non-collagenous protein synthesis during the pressurization period (P < 0.05). The collagenous protein synthesis was not affected by the intermittent sub ambient pressure regimen used in this study. After the intermittent sub-ambient pressurization, the metabolic activities of the chondrocytes returned to normal (control level). The intermittent sub-ambient pressure also produced an increase in the mRNA signals for aggrecan. Therefore, we conclude that intermittent sub ambient pressure may be one of the potential mechanical stimulators of chondrocytes in articular cartilage during dynamic compression. PMID- 10367017 TI - Tensegrity and mechanoregulation: from skeleton to cytoskeleton. AB - OBJECTIVE: To elucidate how mechanical stresses that are applied to the whole organism are transmitted to individual cells and transduced into a biochemical response. DESIGN: In this article, we describe fundamental design principles that are used to stabilize the musculoskeletal system at many different size scales and show that these design features are embodied in one particular form of architecture that is known as tensegrity. RESULTS: Tensegrity structures are characterized by use of continuous tension and local compression; architecture, prestress (internal stress prior to application of external force), and triangulation play the most critical roles in terms of determining their mechanical stability. In living organisms, use of a hierarchy of tensegrity networks both optimizes structural efficiency and provides a mechanism to mechanically couple the parts with the whole: mechanical stresses applied at the macroscale result in structural rearrangements at the cell and molecular level. CONCLUSION: Due to use of tensegrity architecture, mechanical stress is concentrated and focused on signal transducing molecules that physically associate with cell surface molecules that anchor cells to extracellular matrix, such as integrins, and with load-bearing elements within the internal cytoskeleton and nucleus. Mechanochemical transduction may then proceed through local stress-dependent changes in molecular mechanics, thermodynamics, and kinetics within the cell. In this manner, the entire cellular response to stress may be orchestrated and tuned by altering the prestress in the cell, just as changing muscular tone can alter mechanical stability and structural coordination throughout the whole musculoskeletal system. PMID- 10367018 TI - Knee cartilage topography, thickness, and contact areas from MRI: in-vitro calibration and in-vivo measurements. AB - OBJECTIVE: This study assessed the three-dimensional accuracy of magnetic resonance imaging (MRI) for measuring articular surface topographies and cartilage thicknesses of human cadaveric knee joints, by comparison with the calibrated stereophotogrammetric (SPG) method. METHODS: Six fresh frozen cadaveric knees and the knees of four volunteers were imaged with a three dimensional spoiled gradient-recalled acquisition with fat suppression using a linear extremity coil in a 1.5 T superconducting magnet. The imaging voxel size was 0.47 x 0.47 x 1.0 mm. Both a manual and a semi-automated segmentation method were employed to extract topographic measurements from MRI. Following MRI, each of the six cadaveric knees was dissected and its articular surfaces quantified using stereophotogrammetry. The MRI surface measurements were compared numerically with the SPG measurements. RESULTS: For six cadaveric knees, the average accuracies of cartilage and subchondral bone surface measurements were found to be 0.22 mm and 0.14 mm respectively and the thickness measurements demonstrated an average accuracy of 0.31 mm. It was found that while most of the error may be attributed to random measurement error, the accuracy was somewhat affected by systematic errors. For each bone of the knee, accuracies were most favorable in the patella, followed by the femur and then the tibia. The more efficient semi-automated method provided equally good and sometimes better accuracies than manual segmentation. CONCLUSIONS: This study demonstrates that clinical MRI can provide accurate measurements of cartilage topography, thickness, contact areas and surface curvatures of the knee. PMID- 10367019 TI - Injury and reconstruction of the anterior cruciate ligament and knee osteoarthritis. AB - OBJECTIVE: The objective of this study was to study injury and reconstruction of the anterior cruciate ligament (ACL) and their effects on knee osteoarthritis. DESIGN: This manuscript discusses the function of knee ligaments, including the basic mechanical properties, the structural properties of their respective bone ligament-bone complexes, as well as their time- and history-dependent viscoelastic characteristics. The in-situ forces in the ACL and its replacement grafts and knee kinematics before and after ACL reconstruction are also examined. RESULTS: A robotic/universal force-moment sensor (UFS) testing system has been developed which offers a unique method in determining the multiple-degree of freedom knee kinematics and in-situ forces in human cadaveric knees. Under a 110 N anterior tibial load we found at flexion angles of 15 degrees or lower, there was a significantly larger in-situ force in the PL bundle (approximately 75 N) of the ACL as compared to the AM bundle (approximately 35 N)(P < 0.05). We also found that a quadruple semitendinosus and gracilis tendon ACL graft may be better at fully restoring in-situ forces for the whole range of knee flexion when compared to a bone-patellar tendon-bone ACL graft. CONCLUSIONS: The robotic/UFS testing system allows us to determine knee kinematics and the in-situ forces in cadaveric knees in a non-invasive, non-contact manner. Additionally, the ability to reproduce kinematics during testing allows us to evaluate ACL and ACL graft function under external and simulated muscle loading conditions. Finally, we can also examine many of the variables of ACL reconstructions that affect knee kinematics and graft forces including graft tensioning, graft type, graft placement and tibial positioning during graft fixation. PMID- 10367020 TI - Changes in biomechanical properties of tendons and ligaments from joint disuse. AB - OBJECTIVE: The purpose of this paper is to review changes in the biomechanical properties of tendons and ligaments from joint disuse. METHOD: We have reviewed 37 experimental studies on joint disuse, which have been carried out with various models of disuse and with various animals. RESULTS: Immobilization of joints has most commonly been used as a model of disuse. Immobilization of the joint deteriorates the mechanical properties of tendons and ligaments, and reduces their cross-sectional area, although there are some differences in the speed of deterioration among tissues. Remobilization returns the mechanical properties once reduced by immobilization to nearly normal quickly, although the structural properties of the bone-ligament-bone complex continue to lag behind those of the controls. Stress deprivation has been regarded as an essential causative factor in joint disuse. Even if joint motion is allowed, stress deprivation rapidly reduces the mechanical properties of the tendon and ligament tissues, and increases the cross-sectional area of them. These effects appear time- and dose dependent. Restressing increases the mechanical properties once reduced by stress deprivation, although it takes much time to completely recover them. The reduction of the ultimate stress may be explained by the reduction of the total area of collagen fibrils in tendon cross-section and the increase of thin and immature fibrils. PMID- 10367021 TI - Molecular biology and biomechanics of normal and healing ligaments--a review. AB - OBJECTIVE: In this review article, we discuss current data and concepts concerning the molecular biology and biomechanics of both normal and healing ligaments in a rabbit model. METHOD: Data is presented from light microscopy, transmission electron microscopy, molecular biology (RT-PCR), and biomechanical measurements (laxity, stress at failure, modulus, and static creep) or normal, pregnant and healing rabbit medial collateral ligaments. RESULTS: 'Flaws' in scar matrix, smaller-than-normal diameter collagen fibrils, and failure of collagen cross-link maturation may be particularly important deficiencies which appear to be related to ligament scar weakness and perhaps to scar creep. The mechanical behaviours of both normal and healing ligaments are altered by relative states of joint motion and normal ligaments are affected by systemic hormones (particularly during pregnancy). DISCUSSION: Molecular analysis of ligaments and ligament scars, combined with ongoing morphological and biomechanical studies of ligament structure and function, will ultimately reveal which factors can be manipulated clinically to optimize the restoration of normal ligament properties after ligament injuries. Further studies on the mechanisms of ligament healing, genetic markers of repair, and gender-specific differences in ligament repair responses are required. PMID- 10367022 TI - Mechanical load stimulates expression of novel genes in vivo and in vitro in avian flexor tendon cells. AB - OBJECTIVE: Our experiments were designed to test the hypothesis that tendon cells might respond differently to applied strain in vitro than in vivo. DESIGN: We tested cells in whole tendons from exercised chickens and from isolated surface (TSC) and internal tendon (TIF) in vitro that were subjected to mechanical strain. We hypothesized that tendon cells differentially express genes in response to mechanical loading in vivo and in vitro. METHODS: We utilized an in vivo exercise model in which chickens were run on a treadmill in an acute loading regime for 1 h 45 min with the balance of time at rest to 6 h total time. Gene expression was analyzed by a differential display technique. In addition, isolated avian flexor digitorum profundus TSC and TIF cells were subjected to cyclic stretching at 1 Hz, 5% average elongation for 6 h, +/- PDGF-BB, IGF-I, TGF beta 1, PTH, estrogen, PGE2, or no drug and/or no load. mRNA was then collected and samples were subjected to differential display analysis. CONCLUSIONS: Load with or without growth factor and hormone treatments induced expression of novel genes as well as some known genes that were novel to tendon cells. We conclude that the study of gene expression in mechanically loaded cells in vivo and in vitro will lead to the discovery of novel and important marker proteins that may yield clues to positive and negative cell strain responses that are protective under one set of conditions and destructive under another. PMID- 10367023 TI - The effect of exercise training programs on bone mass: a meta-analysis of published controlled trials in pre- and postmenopausal women. AB - With the aging of the population, the medical and social costs of skeletal fragility leading to fractures will cause an immense burden on society unless effective prophylactic and therapeutic regimens can be developed. Exercise is suggested as a possible regimen against involutional bone loss. The purpose of the present meta-analysis is to address a quantitative review of the randomized controlled trials (RCTs) and nonrandomized controlled trials (CTs) on the effects of exercise training programs on bone mass, measured as bone mineral density (BMD) or bone mineral content (BMC), of the lumbar spine (LS) and the femoral neck (FN) in pre- and postmenopausal women. The literature from 1966 through December 1996 was searched for published RCTs and CTs. Study treatment effect is defined as the difference between percentage change in bone mass per year in the training group and the control group. Overall treatment effects (OTs) with the 95% confidence intervals of these study treatment effects were calculated using inverse-variance weighting. Of the 62 articles identified, 25 met the inclusion criteria and were maintained for further analyses. The weighted OTs for the RCTs showed very consistently that the exercise training programs prevented or reversed almost 1% of bone loss per year in both LS and FN for both pre- and postmenopausal women. The two OTs that could be calculated for strength training programs did not reach significance. The OTs for the CTs were almost twice as high as those for the RCTs, which gives an indication of the confounding introduced by the nonrandom allocation of the subjects to groups. PMID- 10367024 TI - Urinary calcium in perimenopausal women: normative values. AB - Twenty-four hour urinary calcium was measured under controlled dietary and metabolic balance conditions in 191 normal perimenopausal women at 5-year intervals, under varying conditions of estrogen status and dietary calcium intake, generating 586 values overall. Upper and lower limits of the normal range for urinary calcium excretion are presented for the estrogen-replete and estrogen deprived states and for varying levels of calcium intake. PMID- 10367025 TI - Absorption of calcium as the carbonate and citrate salts, with some observations on method. AB - Calcium supplement use has increased and there is confusion about the relative absorbability of various sources. Absorbability of calcium from the carbonate and citrate salts was compared at 300 mg and 1000 mg calcium loads, ingested as part of a light breakfast meal. Absorption was measured at the high load both by tracer appearance in serum and by the absorptive increment in urinary calcium, and at the low load by the tracer method only. Subjects were 37 healthy adult men and women, studied as outpatients, and each tested on both salts at the same load. Mean tracer absorption (+/- SD) for both salts combined was 36.0% at the 300 mg load and 28.4% at the 1000 mg load. In both experiments the observed mean difference in absorption between salts was very small. By the tracer method the within-subject difference (carbonate less citrate) was +3.3% +/- 1.2% of the ingested dose (mean +/- SEM; P < 0.05) at the high load, and at the low load, 3.6% +/- 2.7% (NS). Combining the two experiments yielded zero difference between sources. By the urinary calcium increment method, the mean difference between salts at the 1000 mg load was 1.8 +/- 4.1 mg (NS). Side-by-side comparisons of the two methods revealed that the tracer method was 3 times more sensitive than the urinary increment method. We conclude that, when taken with food, calcium from the carbonate salt is fully as absorbable as from the citrate, and that the urinary increment method is not sufficiently sensitive to be useful in comparing sources in free-living subjects. PMID- 10367026 TI - Stiffness in discrimination of patients with vertebral fractures. AB - We measured the ultrasound parameters of the heels of 49 women with vertebral fractures and 87 age-matched controls using an Achilles ultrasound device. Average broadband ultrasound attenuation (BUA), speed of sound (SOS) and Stiffness were significantly lower in fracture patients (p < 0.0001). We also estimated the ultrasound parameters of patients compared with age-matched non fracture controls and found the mean BUA to be -1.02 SD below control values. The mean SOS was -0.97 SD and the mean Stiffness was -1.12 SD below control values. Femoral bone mineral density (BMD) at the neck, Ward's triangle and the trochanter, the total-body BMD and L2-4 BMD were measured with dual-energy X-ray absorptiometry (DXA) and found to be significantly lower in fracture patients (p < 0.0001). All correlation coefficients between ultrasound parameters and DXA measurements were > 0.5 and statistically significant (p < 0.0001). A stepwise logistic regression with presence or absence of vertebral fracture as the response variable and all ultrasound--DXA parameters as the explanatory variables indicated that the best predictor of fracture was Stiffness, with additional predictive ability provided by spine BMD. Sensitivity and specificity of all measures were determined by the areas under the receiver operating characteristic (ROC) curve, which were 0.76 +/- 0.04 for BUA, 0.77 +/- 0.04 for SOS, 0.78 +/- 0.04 for Stiffness and 0.78 +/- 0.03 for spine BMD. The areas under the ROC curves of BUA, SOS, Stiffness and spine BMD were compared and it was found that Stiffness and spine BMD were significantly better predictors of fracture than BUA and SOS. These results support many recent studies showing that ultrasound measurements of the os-calcis have diagnostic sensitivity comparable to DXA, and also demonstrated that Stiffness was a better predictor of fracture than spine BMD. PMID- 10367027 TI - Fracture incidence in Olmsted County, Minnesota: comparison of urban with rural rates and changes in urban rates over time. AB - Using the data resources of the Rochester Epidemiology Project, we carried out a descriptive study of fracture incidence among the residents of Olmsted County, Minnesota. During the 3-year period 1989-91, 2901 County residents > or = 35 years of age experienced 3665 separate fractures. The age- and sex-adjusted (to 1990 United States whites) incidence of any fracture was 2205 per 100,000 person years (95% CI, 2123 to 2286) and that of all fractures was 2797 per 100,000 (95% CI, 2705 to 2889). Age-adjusted fracture rates were 40% greater among women. Incidence rates increased with age in both sexes. One-third of the fractures involved the hip, spine or distal forearm - the skeletal sites traditionally associated with osteoporosis. The age- and sex-adjusted incidence of fractures due to moderate trauma (2205 per 100,000 person-years; 95% CI, 2106 to 2303) was twice that of fractures due to more severe trauma (1164 per 100,000; 95% CI, 1106 to 1223) and 12 times that of pathological fractures (178 per 100,000; 95% CI, 133 to 222). Overall fracture rates were 15% greater among residents of the central city of Rochester compared with the rural portion of Olmsted County. The incidence of limb fractures among Rochester residents was 14% higher than comparable rates documented for this community 20 years earlier in 1969-71, due mainly to a substantial increase in the incidence of leg fractures. PMID- 10367028 TI - Development and evaluation of a phantom for morphometric X-ray absorptiometry. AB - Morphometric X-ray absorptiometry (MXA) provides the potential to assess vertebral deformity using a technique with much lower radiation dose to the patient than standard radiographic procedures. MXA overcomes many other limitations such as cone beam distortion observed in conventional plane radiographs. A phantom has been designed to assess the accuracy of the MXA technique, to monitor long-term precision and to assess inter- and intra-operator variability. The phantom consists of two columns of 12 cylinders representing the vertebral bodies, one of regular components and one representing vertebral deformities. Each column may be inserted, as required, into a Perspex torso mimicking block. Initial assessment on the Lunar Expert-XL demonstrates that the phantom provides image parameters reflecting those found clinically. Measurement of vertebral height was found to be consistently underestimated by 4.9%. Operator precision ranged from 0.6% for posterior height measurement to 1.0% for middle height measurement of the regular component column. The corresponding precision range for the column representing vertebral deformation was 0.6% (posterior) and 1.1% (middle). Analysis of 10 scans of each column by two independent operators demonstrated a few significant differences in height assessment confined to the 'thoracic' region of the regular column. However, inter-operator variability was found to increase with increasing complexity of vertebral shape producing several differences, particularly in posterior height assessment of the deformed column. PMID- 10367029 TI - Risk factors for hip fracture in men from southern Europe: the MEDOS study. Mediterranean Osteoporosis Study. AB - The aims of this study were to identify risk factors for hip fracture in men aged 50 years or more. We identified 730 men with hip fracture from 14 centers from Portugal, Spain, France, Italy, Greece and Turkey during the course of a prospective study of hip fracture incidence and 1132 age-stratified controls selected from the neighborhood or population registers. The questionnaire examined aspects of work, physical activity past and present, diseases and drugs, height, weight, indices of co-morbidity and consumption of tobacco, alcohol, calcium, coffee and tea. Significant risk factors identified by univariate analysis included low body mass index (BMI), low sunlight exposure, a low degree of recreational physical activity, low consumption of milk and cheese, and a poor mental score. Co-morbidity including sleep disturbances, loss of weight, impaired mental status and poor appetite were also significant risk factors. Previous stroke with hemiplegia, prior fragility fractures, senile dementia, alcoholism and gastrectomy were associated with significant risk, whereas osteoarthrosis, nephrolithiasis and myocardial infarction were associated with lower risks. Taking medications was not associated with a difference in risk apart from a protective effect with the use of analgesics independent of co-existing osteoarthrosis and an increased risk with the use of anti-epileptic agents. Of the potentially 'reversible' risk factors, BMI, leisure exercise, exposure to sunlight and consumption of tea and alcohol and tobacco remained independent risk factors after multivariate analysis, accounting for 54% of hip fractures. Excluding BMI, 46% of fractures could be explained on the basis of the risk factors sought. Of the remaining factors low exposure to sunlight and decreased physical activity accounted for the highest attributable risks (14% and 9% respectively). The use of risk factors to predict hip fractures had relatively low sensitivity and specificity (59.6% and 61.0% respectively). We conclude that lifestyle factors are associated with significant differences in the risk of hip fracture. Potentially remediable factors including a low degree of physical exercise and a low BMI account for a large component of the total risk. PMID- 10367030 TI - The association of bone mineral density with vitamin D receptor gene polymorphisms. AB - A recent meta-analysis of 16 publications suggested that bone mineral density (BMD) is not associated with vitamin D receptor (VDR) gene polymorphism (VDRGP) at the 0.05 significance level when a study with genotyping mistakes is excluded. We wished to determine whether 'positive' findings supporting the BMD-VDRGP association may be explained by chance, and what factors affect the outcomes of these studies. Seventy-five articles and abstracts on the association of VDRGP with BMD and related skeletal phenotypes published before January 1997 were identified. Twenty-three of 67 (34.3%) studies on spinal BMD and 22 of 51 (43.1%) on femoral neck BMD had found a BMD-VDRGP association at p < 0.05, significantly (p = 7 x 10(-14) for spinal BMD, p = 9 x 10(-16) for hip BMD) higher than the expected 5% false positive rate under the null hypothesis of 'no association'. 'Positive' results were more frequently observed in studies on females before the menopause than those on females after the menopause (p < 0.02) or on male and female subjects combined (p < 0.05) when skeletal phenotypes at any bone sites were considered. The 'positive rate' among studies was also influenced by the age range of subjects studied and by the inclusion of subjects with osteoporosis. It is concluded that: (1) BMD is associated with VDRGP with high levels of confidence and (2) non-genetic factors and genetic heterogeneity interfere with the detection of the effects of VDRGP on bone phenotypes. PMID- 10367031 TI - In vivo MRI measurements of bone quality in the calcaneus: a comparison with DXA and ultrasound. AB - Magnetic resonance imaging (MRI) has shown promise in the assessment of bone architecture. The precision and feasibility of MRI measurements in osteoporosis in vivo have been assessed in this study. T2' was calculated from measurements of T2 and T2* in the calcaneus of 32 postmenopausal women using a gradient-echo sequence PRIME (Partially Refocused Interleaved Multiple Echo). This sequence allows the measurement of T2 and T2* in one acquisition. In vivo measurements of bone mineral density (BMD) by dual-energy X-ray absorptiometry (DXA) were made in the calcaneus, spine and femoral neck. The ultrasound parameters broadband ultrasound attenuation (BUA) and speed of sound (SOS) were also measured in the calcaneus. These three techniques have not previously been compared in the same study population. The precision of the MRI technique was poor relative to the DXA and ultrasound techniques, with a CV of 6.9% +/- 4.4% for T2' and 5.5% +/- 3.6% for T2*. Approximately 4% of this is due to system error as determined by phantom measurements. The postmenopausal women were classified as having low BMD if they had a lumbar spine (L2-4) BMD of less than 0.96 g/cm2 (more than 2 standard deviations below normal peak bone mass). Calcaneal T2' was significantly correlated with calcaneal BMD (r = -0.79, p < 0.0001), BUA (r = -0.59, p = 0.0004) and SOS (r = -0.58, p = 0.0006). T2' was significantly different in postmenopausal women with normal BMD and those with low BMD (p < 0.01). However, the difference was of only borderline significance (p < 0.06) after adjustment for age and years since menopause. PMID- 10367032 TI - Prophylactic use of alfacalcidol in corticosteroid-induced osteoporosis. AB - One hundred and forty-five patients suffering from diseases requiring long-term treatment with high doses of corticosteroids (30 mg/day or greater of prednisolone) were recruited to the study. Patients had to be steroid naive on entry to the study (not more than 15 days of treatment with a corticosteroid within the previous 24 months). Patients were randomized to receive either 1 microgram/day alfacalcidol or placebo capsules for 12 months. Bone mineral density (BMD) of the lumbar spine was assessed by dual-photon absorptiometry on entry and after 3, 6 and 12 months' treatment. Safety was monitored by the recording of all adverse events reported by patients and the regular screening of blood samples for hematology and serum biochemistry. Of the 145 patients, 74 were randomized to alfacalcidol and 71 to placebo. The treatment groups were well matched at baseline with no significant differences in demographic, clinical or biochemical parameters. The mean equivalent dose of prednisolone at baseline was 46.6 mg/day and 46.3 mg/day for the alfacalcidol and placebo group respectively. From the 145 patients randomized to treatment, 71 (38 who received alfacalcidol and 33 who received placebo) provided BMD data both at baseline and at 3, 6 and 12 months. The percentage change in BMD after 6 months' treatment was -2.11% in the alfacalcidol group and -4.00% in the placebo group (p = 0.39). After 12 months the percentage change in BMD was +0.39% (CI: -4.28 to 4.81) in the alfacalcidol group and -5.67% (CI: -8.13 to -3.21) in the placebo group, this difference (6.06%, CI: 0.88 to 11.24) being statistically significant (p = 0.02). An intention to treat analysis also showed a significant difference between the two treatment groups in alfacalcidol's favor (3.81%, p = 0.01; CI: 0.92 to 6.70). There was no significant difference between the two treatment groups in the corticosteroid dose at any time point during the study. Serum calcium was measured throughout and there were no significant differences between the two treatment groups at any visit. This study suggests that alfacalcidol can prevent corticosteroid-induced bone loss from the lumbar spine. Long-term use of alfacalcidol was not associated with any significant adverse effects in this diverse group of patients. PMID- 10367034 TI - Case-control study of the pathogenesis and sequelae of symptomatic vertebral fractures in men. AB - To investigate the pathogenesis and sequelae of symptomatic vertebral fractures (VF) in men, we have performed a case-control study, comparing 91 men with VF (median age 64 years, range 27-79 years) with 91 age-matched control subjects. Medical history, clinical examination and investigations were performed in all patients and control subjects, to identify potential causes of secondary osteoporosis, together with bone mineral density (BMD) measurements. BMD was lower at the lumbar spine and all sites in the hip in patients with VF than in control subjects (p < 0.001). Potential underlying causes of secondary osteoporosis were found in 41% of men with VF, compared with 9% of control subjects (OR 7.1; 95% CI 3.1-16.4). Oral corticosteroid and anti-convulsant treatment were both associated with a significantly increased risk of VF (OR 6.1; 95% CI 1.3-28.4). Although hypogonadism was not associated with an increased risk of fracture, the level of sex hormone binding globulin was higher (p < 0.001) and the free androgen index lower (p < 0.001) in men with VF than control subjects. Other factors associated with a significantly increased risk of VF were family history of bone disease (OR 6.1; 95% CI 1.3-28.4), current smoking (OR 2.8; 95% CI 1.2-6.7) and alcohol consumption of more than 250 g/week (OR 3.8; 95% CI 1.7 8.7). Men with VF were more likely to complain of back pain (p < 0.001) and greater loss of height (p < 0.001) than control subjects, and had poorer (p < 0.001) scores for the energy, pain, emotion, sleep and physical mobility domains of the Nottingham Health Profile. We conclude that symptomatic VF in men are associated with reduced BMD, underlying causes of secondary osteoporosis such as corticosteroid and anti-convulsant treatment, family history of bone disease, current smoking and high alcohol consumption, and that they impair the perceived health of the individual. PMID- 10367033 TI - Evaluation of finger ultrasound in the assessment of bone status with application of rheumatoid arthritis. AB - Osteoporosis associated with active rheumatoid arthritis (RA) has been demonstrated in both the axial and peripheral skeleton, especially the periarticular regions more directly affected by the disease. Quantitative ultrasound (QUS) is a recently accepted tool for the assessment of bone status, and therefore could be used to monitor bone changes in RA patients. In a cross sectional study we measured ultrasound velocity (Ad-SOS) through the proximal phalanges in three groups of female subjects. These included: 51 patients with rheumatoid arthritis (group 1), 44 general practitioner (GP)-referred patients for osteopenia (group 2) and 52 young healthy volunteers (group 3). For groups 1 and 2 bone mineral density (BMD) of the lumbar spine and proximal femur were also measured. For the RA patients BMD of the hand, measurement of hand function (HAQ and grip strength) and disease activity (ESR and CRP) were also assessed. The precision of long-term Ad-SOS measurements on volunteers gave a root mean square coefficient of variation (CV) of 0.7% and standardized CV of 3.6%. No statistically significant effect of dominance was observed in the measured Ad-SOS between the dominant and non-dominant hand (r = 0.96, p < 0.001). Ad-SOS was found to be significantly different in the three groups (p < 0.0001). Ad-SOS was highly dependent on age (r = -0.67), with a gradual reduction (-5.2 m/s per year) after the age of 30 years for female patients in both group 1 and group 2. Ad-SOS was significantly correlated with lumbar spine, femoral neck and hand BMD, with correlation coefficients of 0.49, 0.51 and 0.72 respectively for RA patients. Finger ultrasound was moderately correlated with measures of hand function, with coefficients of 0.37 and 0.39 for HAQ and grip strength respectively. Hand BMD also correlated to the same power with these parameters. Neither finger ultrasound nor BMD was significantly correlated with ESR and CRP (measures of disease activity). We have demonstrated that bone status can be assessed quickly and cheaply using a portable QUS device. Ad-SOS relates to the measure of hand function in RA patients. Longitudinal studies are required to determine the usefulness of finger ultrasound for monitoring disease progression or the effect of treatment in RA. PMID- 10367035 TI - Ultrasound velocity and attenuation in cancellous bone samples from lumbar vertebra and calcaneus. AB - We report a study of ultrasound velocity and broadband ultrasound attenuation (BUA) in human cancellous bone samples. The influence of density and microarchitecture on ultrasound propagation in cancellous bone was examined. A total of 20 samples from vertebra L1 and 21 from calcanei were studied. The direction of ultrasound propagation was anteroposterior in the vertebra and lateromedial in the calcaneus. The relationships between ultrasonic parameters and density of bone samples, apparent ash density, trabecular bone volume (BV/TV) and trabecular thickness (Tb.Th) were analyzed using a simple linear model and a multiple regression model. Velocity of ultrasound and BUA were positively correlated with density and morphometric parameters, in both vertebra and calcaneus. The best correlation was found between velocity and bone sample density in vertebra (r = 0.961, p < 0.0001) and the worst between velocity and trabecular thickness in calcaneus (r = 0.632, p = 0.002). The best correlation for BUA was with BV/TV in vertebra (r = 0.960, p < 0.0001). Using the stepwise regression procedure, BV/TV only was selected as significant for BUA and apparent ash density with Tb.Th for velocity, in both vertebra and in calcaneus. The possible influence of trabecular configuration on ultrasonic parameters is discussed, emphasizing the different slopes of regression lines obtained for vertebra and calcaneus, sites with different architecture of trabecular bone. PMID- 10367036 TI - Whole-body bone mineral measurements in 15-year-old Swedish adolescents. AB - Bone mineral area (BA), total bone mineral content (TBMC) and total bone mineral density (TBMD) were assessed by dual-energy X-ray absorptiometry (DXA) in 396 randomly selected, healthy 15-year-old Swedish boys and girls. The influence of body size, pubertal development, physical activity level (PAL), total energy expenditure (TEE), dietary intake of energy, calcium and vitamin D, and alcohol and smoking habits on TBMC and TBMD were examined in bi- and multivariate analyses. In bivariate analyses BA, TBMC and TBMD showed strong correlations with weight, height and TEE in both sexes. In boys but not in girls these bone variables were significantly correlated with dietary intakes of energy, calcium and vitamin D. No significant correlations were found between PAL and the three bone variables. In multivariate analyses with TBMC as dependent variable BA, height, weight and Tanner stages explained 88% and 87% of the variance in boys and girls respectively. In similar analyses with TBMD as dependent variable the corresponding figures were 50% and 54%. The major part of the variance in all these models was explained by BA, and only a few percent by all the other independent variables. No significant reduction was found when TEE or daily intakes of calcium or vitamin D were introduced into the models. These results illustrate the importance of including BA, weight and height as independent variables in regression models of TBMC to avoid spurious associations with other variables in the analyses. The results may also indicate that in normal Swedish adolescents environmental factors such as dietary intake of nutrients play a minor role as determinants of bone mineralization. High levels of physical activity and bone mineral measures possibly explain the lack of significant correlations between these variables and do not imply a lack of association. PMID- 10367037 TI - An unexpected change in DXA calibration not detected by routine quality control checks. AB - Since its commercial introduction a decade ago, the technique of dual-energy X ray absorptiometry (DXA) has been widely recognized as a useful and sensitive method of measuring changes in bone mineral density (BMD) at selected sites in the skeleton such as the spine and proximal femur. Because of their high precision and stable calibration, DXA scanners are frequently used in clinical trials to evaluate new treatments for osteoporosis. Quality assurance procedures based on regular scanning of phantoms are widely adopted in such trials, and continuity of the phantom BMD measurements is generally believed to ensure continuity in the in-vivo calibration. We report a change in calibration of a DXA scanner that occurred during a clinical trial where the calibration shift was different for the spine and femur sites and was not predicted or explained by the standard quality control procedures using phantoms. However, we show that provided patients enrolled in studies are thoroughly randomized and the statistical analysis is confined to the differences between the treated and control groups, then the effects of such calibration shifts on conclusions regarding the efficacy of treatment are considerably smaller than the random statistical errors. PMID- 10367038 TI - High prevalence of low femoral bone mineral density in elderly women living in nursing homes or community-dwelling: a plausible role of increased parathyroid hormone secretion. AB - The present study was designed to visit elderly women living in nursing homes and to compare their femoral neck bone mineral density (BMD) and circulating levels of parathyroid hormone (PTH) and 25-OH vitamin D (25-OHD) with those of subjects living at home, in the immediate vicinity of the nursing homes. Of 1483 women, aged 70 years and older, who were selected, 993 agreed to participate in this trial. Their femoral neck BMD (n = 993) was measured by dual-energy X-ray absorptiometry, with a specific device installed in a mobile truck. The circulating levels of 25-OHD and PTH were assessed after an overnight fast (n = 748). After stratification for age, there were no significant differences in mean femoral neck BMD values, prevalence of femoral neck osteoporosis, mean serum 25 OHD and prevalence of absolute or relative 25-OHD deficiency between the two groups. Serum levels of PTH were significantly higher in women over 80 years old living in nursing homes, compared with the community-dwelling women. After adjustment for age, a significant relation was found between femoral neck BMD and PTH levels in the whole population (p = 0.004) and in community-dwelling subjects (p = 0.039). When stratifying our population by quartiles of serum PTH values, the odds ratios for femoral neck osteoporosis were significantly increased for the top two quartiles compared with the lowest one both before (p = 0.00146) and after (p = 0.0013) adjustment for age and type of housing. From this study we conclude that femoral osteoporosis is largely underestimated in European women. Living in a nursing home is not, per se, a risk factor for decreased femoral BMD, and circulating PTH levels are a key determinant of low femoral bone density and osteoporosis. PMID- 10367039 TI - Usefulness of armspan and height comparison in detecting vertebral deformities in women. AB - The prevalence of vertebral fractures in women increases with age but only about one third of these fractures are symptomatic. On the other hand, the presence of vertebral fractures is an independent risk factor for new osteoporotic fractures. In the present study we examined the hypothesis that differences between armspan and height are related to the presence of vertebral deformities in a cohort of 494 women aged between 55 and 84 years (mean age 67.6 years, SD 8.2 years) who were randomly selected from a large general practice in The Netherlands. Height and armspan were measured and vertebral morphometry of lateral radiographs of the spine was performed. Both height and armspan decreased significantly with age. The correlation between armspan and height was 0.83. Vertebral deformities were present in 32.7% of the women (grade I in 22.4% and grade II in 10.3%). Only the prevalence of grade II deformities rose with age. The variation of the difference between armspan and height in the groups with or without grade II vertebral deformities was comparable and relatively large (range > 15 cm). The difference in mean values was small between those groups (1.6 cm) and could not differentiate between women with and without vertebral deformities. Our data show that the presence of vertebral deformities cannot be detected by the difference between armspan and height. PMID- 10367040 TI - Polymorphism of the vitamin D receptor gene and corticosteroid-related osteoporosis. AB - Corticosteroid therapy (CST) is associated with reduced intestinal calcium absorption, bone loss and increased fracture risk. As polymorphisms of the vitamin D receptor (VDR) gene may be associated with bone mineral density (BMD) and intestinal calcium absorption, we asked whether patients with a given VDR genotype receiving CST may be at increased or decreased risk for corticosteroid related bone loss and osteoporosis. We measured areal BMD (g/cm2) by dual-energy X-ray absorptiometry in 193 women (50 premenopausal, 143 postmenopausal) and 70 men with rheumatoid arthritis (n = 44), obstructive airway diseases (n = 128) and other corticosteroid-treated diseases (n = 91). All patients received a cumulative dose greater than 1.8 g per year or a minimum of 5 mg daily of prednisolone or equivalent for at least 1 year. VDR alleles were typed by polymerase chain reaction assay based on the polymorphic BsmI and TaqI restriction sites. BMD in patients was expressed as a Z-score (mean +/- SEM) derived from age- and gender-matched controls. BMD was reduced in patients at the lumbar spine (bb, -0.52 +/- 0.12; Bb, -0.47 +/- 0.11; BB, -0.65 +/- 0.18 SD; p < 0.01), femoral neck (bb, -0.46 +/- 0.10; Bb, -0.34 +/- 0.10; BB, -0.54 +/- 0.14 SD; p < 0.01), Ward's triangle (bb, -0.44 +/- 0.10; Bb, -0.31 +/- 0.10; BB, -0.45 +/- 0.13 SD; p < 0.01), and trochanter (bb, -0.50 +/- 0.10; Bb, -0.30 +/- 0.10; BB, -0.44 +/- 0.14 SD; p < 0.01). However, there was no significant difference in the deficit in BMD in any of the genotypes, either before or after adjusting for age, sex, body mass index, disease type, age at onset of disease, disease duration, cumulative steroid dosage, smoking status and dietary calcium intake. Similarly, there were no detectable differences between the BsmI genotypes and the rate of bone loss in 79 patients with repeated BMD measurements at an interval of 4-48 months. The data suggest that the VDR genotypes may not be a means of identifying patients at greater risk of corticosteroid-related bone loss. PMID- 10367041 TI - Attitudes to osteoporosis and hormone replacement therapy among elderly women. AB - This study compares the attitudes toward osteoporosis and its treatment between a group of elderly women admitted to hospital for therapy of an osteoporotic fracture and a control group admitted for joint replacement surgery. We surveyed 97 women (64 with a fracture, and 33 controls) and found that the two groups of patients demonstrated a similar risk factor profile for osteoporosis and poor knowledge of osteoporosis and its available treatments, including hormone replacement therapy (HRT). By selecting a control group of women with no recent fracture, we hoped to highlight the effect of sustaining a recent fracture on attitudes to treatment. Initially only 10% (8 in the fracture group and 2 in the control group) were interested in treatment for osteoporosis, but those women who had been admitted with a fracture were significantly more receptive to education about osteoporosis and to the offer of further investigation and treatment of osteoporosis (38 versus 10, p = 0.007). We conclude that it is worthwhile offering education, screening and treatment to elderly patients who present with a fracture. PMID- 10367042 TI - A secular trend in hip fracture incidence in East Germany. AB - The central Inpatient Register of the former German Democratic Republic was used to study the population-based epidemiology of hip fractures among 16.5 million East Germans. Incidence rates for hospital discharges for proximal femoral fractures for the age group 60 years and over were calculated for the years 1971 to 1989, the year before unification. Incidence rates for 1989 are similar to figures reported from the UK and The Netherlands, but lower than Scandinavian rates. A decrease in the admission rate was noted from 1971 to 1974 of 4.5% each year on average, and an increase from 1974 onwards of 4.4% on average. This change was observed to a different extent in all age groups. The female:male ratio of the standardized discharge incidence was stable at 2.3:1 and the female:male ratio of manifest cases increases from 4.1:1 in 1971 to 5.1:1 in 1989. An exponential increase in the incidence rates was observed with age. This apparent rate overestimated both the rate for true incident cases (by about 25 30%, if adjustments are made for readmissions and transfers) and their trend. Adjusted estimates for incident fractures show an increase of 2% annually. Cohort effects due to changed selective forces appear to be one reasonable causal explanation. PMID- 10367043 TI - The prevalence of osteoporosis in nursing home residents. AB - This study describes the prevalence of osteoporosis in a statewide sample of nursing home residents. Composite forearm bone mineral density (BMD) (including the distal radius and the distal ulna) of 1475 residents aged 65 years and older from 34 randomly selected, stratified nursing homes was assessed. BMD was expressed with reference to World Health Organization diagnostic criteria. Trends with age, gender and race were consistent with other populations. However, prevalence estimates were higher than community-based age-specific rates. The prevalence of osteoporosis for white female residents increased from 63.5% for women aged 65-74 years to 85.8% for women over 85 years of age. Only 3% had composite forearm BMD within 1 standard deviation of the young adult mean. The significance of the high prevalence of low BMD in nursing home residents is the increased fracture risk it may confer. In community cohorts of white women, the risk of hip fracture increases approximately 50% for every 1 standard deviation decrease in bone mass. However, the degree to which BMD contributes to fracture risk in this population has not been well established. PMID- 10367044 TI - Influence of current and past hormone replacement therapy on bone mineral density: a study of discordant postmenopausal twins. AB - There is controversy about the ideal timing of hormone replacement therapy (HRT) and duration of treatment. In this study we have examined intrapair differences in bone mineral density (BMD) in twins who were discordant for HRT use. Twin pairs in which only one co-twin had been exposed to HRT for more than 12 months continuously were selected from 365 postmenopausal monozygotic (MZ) and dizygotic (DZ) pairs recruited as part of the St Thomas' Adult UK Twin Registry of normal volunteers. BMD was measured by dual-energy X-ray absorptiometry at the lumbar spine and femoral neck. Intrapair differences in BMD between HRT users and non users were compared. A total of 65 HRT-discordant pairs were identified, of which 36 were discordant for current HRT use (mean age: 55.3 years, median duration of HRT use: 36 months) and 29 were discordant for past HRT use (mean age: 60.4 years, median HRT duration: 30 months). Among current users BMD was consistently and significantly higher than in non-users at both sites (lumbar spine mean intrapair difference (IPD%): 12.3%, 95% confidence interval (CI): 7.1%, 17.5%; femoral neck IPD%: 8.6%, 95% CI: 3.4%, 13.7%). The intrapair differences were substantially smaller when past users and non-users were compared (lumbar spine IPD%: 2.4%, 95% CI: -3.7%, 8.6%; femoral neck IPD%: 0.4%, 95% CI: -5.3%, 6.0%). These differences remained little changed after adjusting for the potential confounding effects of the duration of HRT use, and intrapair differences in alcohol and tobacco consumption and physical exercise. The results confirm, in a closely matched design, the findings of other observational research that current use of HRT has a major effect on BMD at the lumbar spine and femoral neck. Past users of HRT do not, however, show the same benefits. The clinical implications of these findings are that HRT needs to be used continuously to influence BMD and that alternative treatments need to be considered in those who discontinue HRT. PMID- 10367045 TI - Treatment with active vitamin D metabolites and concurrent treatments in the prevention of hip fractures: a retrospective study. AB - The purpose of this study was to determine the effect of treatment with active vitamin D metabolites and other concurrent medication on the prevention of hip fractures in elderly women. We inspected the medical records of the entire female population over 65 years of age on Sado Island, and followed a total of 11,377 women for a 3-year period. Of these, 1208 osteoporotic patients were treated with either 1,25-(OH)2D3 or 1 alpha-(OH)D3. The 765 patients who received the minimum effective dosage for more than 6 months made up the 'treatment group'. Nearly half these patients were also treated with either calcitonin or calcium. The 443 patients who received treatment with active vitamin D metabolites, but at a dosage or for a duration that did not meet the criteria for the treatment group, were deemed the 'ineffective group'. The remaining 10,169 women were the 'non treatment group'. Fractures in the non-treatment group occurred at a rate of 39.8 fractures/10,000 person-years. The rate in the treatment group was 10.8, which was significantly lower (p = 0.039). Interestingly, the fracture rate after ceasing treatment was 52.1, which was significantly higher (p = 0.002) than the rate in patients receiving treatment. No statistical differences in the fracture rate were found between the ineffective, non-treatment and post-treatment groups. A reduction in the fracture rate was observed only in the treatment subgroup that did not also receive calcitonin (p = 0.042), and not in the subgroup that also received calcitonin therapy (p = 0.333). However, there was no statistical difference in the hip fracture rates between these two subgroups (p = 0.157) and the actual number of fractures was minimal (0 vs 2). Therefore, in this study, the advantage of treatment with active vitamin D alone over combined treatment with calcitonin seems to be marginal. IN CONCLUSION: (1) treatment with active vitamin D metabolites and with combined therapy may be marginally effective in preventing hip fractures, and (2) stopping the treatment clearly increases the risk of hip fractures. PMID- 10367046 TI - Therapy of established postmenopausal osteoporosis with monofluorophosphate plus calcium: dose-related effects on bone density and fracture rate. AB - Recent experience from different groups suggests that low fluoride doses resulting in moderate increases in bone mineral density (BMD) may be advantageous in terms of fracture-reducing potency. In a randomized prospective 3-year study we examined the therapeutic efficacy of different dosages of monofluorophosphate (MFP) plus calcium in comparison with calcium alone in 134 women with established postmenopausal osteoporosis (mean age 64.0 years, average vertebral fractures per patient 3.6). Group A received 1000 mg calcium/day and a low-dose intermittent MFP regimen (3 months on, 1 month off) corresponding to an average daily fluoride ion dose of 11.2 mg. Group B received 1000 mg calcium/day plus continuous MFP corresponding to 20 mg fluoride ions per day. Group C was treated with 1000 mg calcium alone throughout the study period. Bone density was measured with dual energy X-ray absorptiometry at L2-4 and three proximal femur areas and with single photon absorptiometry at two radius sites. New vertebral fractures were identified from annual lateral radiographs of the spine. A significant reduction in subjective complaints as measured by a combined pain-mobility score (CPMS) was found in both fluoride groups in comparison with the calcium monotherapy group. Group A showed increases in BMD at all six measuring sites, reaching +12.6% at the spine after 3 years. In group B we found significant increases at the spine, Ward's triangle and distal radius, but slight decreases at the femoral neck and radius shaft. For the spine the average change amounted to +19.5% after 3 years. In group C losses of BMD were observed at all six sites, with an average loss of 1.6% for the spine at the end of the study. The incidence of new vertebral fractures per 100 patient-years was 8.6, 17.0 and 31.6 in groups A, B and C, respectively. In conclusion, both calcium-MFP regimens resulted in significantly lower vertebral fracture rates than calcium monotherapy. However, the low intermittent MFP regimen, leading to a mean annual increase in spinal BMD of only 4.2%, showed a clear trend to greater effectiveness in reducing vertebral fracture than the higher fluoride dosage that was followed by an average spinal BMD increase of 6.5% per year. Furthermore the rate of fluoride-specific side effects (lower-extremity pain syndrome) was 50% lower in patients receiving the lower fluoride dosage. PMID- 10367047 TI - Normal changes in spinal bone mineral density in a Chinese population: assessment by quantitative computed tomography and dual-energy X-ray absorptiometry. AB - This study was designed to determine age- and gender-based normative values for spinal bone mineral density (BMD) in a Chinese population. In addition, we compared our data with those of other countries and populations. Four hundred and forty-three healthy Chinese subjects, aged 10-79 years (189 males, mean age 46.9 years; 254 females, mean age 45.7 years) were recruited for BMD assessment. BMD was measured by quantitative computed tomography (QCT) and dual-energy X-ray absorptiometry (DXA), including posteroanterior DXA (PA-DXA), lateral DXA (L-DXA) and midlateral DXA (mL-DXA). For both genders, BMD values peaked in the 10-19 year age group when measured by QCT, and in the 30-39 year age group when measured by PA-DXA. BMD values decreased with age after reaching peak bone density in males and females for all measurements, except for PA-DXA in males. Male BMD values by DXA tended to increase beginning with the 60-69 age group through the 70-79 age group whether by PA-DXA, or L-DXA and mL-DXA. However, male QCT data showed stable BMD values among these two older groups. Comparative results showed female QCT data were higher in the 20-39 age group and lower after the 40-49 age group compared with American females. The peak BMD value by PA-DXA in Chinese females was reached in the same age group as American and European females and was similar in magnitude (p > 0.05). However, the peak BMD value for Chinese females was reached earlier and was significantly higher than that observed in Japanese females (p < 0.001). We conclude that the age group in which the peak BMD values are reached is different depending on the technique used, as is the calculated age-related rate of bone loss. It can be speculated that such differences reflect different timing for bone maturation in cancellous and cortical bone. PMID- 10367048 TI - Separate and combined value of bone mass and gait speed measurements in screening for hip fracture risk: results from the EPIDOS study. Epidemiologie de l'Osteoporose. AB - Based on data from the EPIDOS prospective study, we have shown that femoral bone mineral density (BMD), calcaneal ultrasound measurements and fall-related factors are significant predictors of the risk of hip fracture. The goal of the present investigation, in the same cohort of elderly women, was (1) to assess and compare the value of femoral BMD, calcaneal broadband ultrasound attenuation (BUA), gait speed and age for identifying elderly women at high risk of hip fracture and (2) to determine whether combining two or more of these measurements would improve predictive ability over single measures. A total of 5895 elderly women had baseline measurements of femoral neck BMD by dual-energy X-ray absorptiometry, calcaneal BUA and gait speed. During an average of 33 months of follow-up, 170 women suffered a hip fracture. We compared the sensitivity and specificity of single and combined measures for three specific cutoff levels to define high risk, i.e., the median, the top quartile and the top decile of risk. We found that femoral BMD, calcaneal BUA, gait speed and age have approximatively the same discriminant value to identify women at high risk of hip fracture even though certain measures and combinations of measures have a significantly higher sensitivity for certain cutoff levels. The sensitivity of the available screening tools is low, even when they are combined: to obtain a sensitivity of about 80%, approximately 50% of the population must be considered to be at high risk. PMID- 10367049 TI - Dichoptically cancelled motion. AB - We sought to determine whether or not motion-from-texture mechanisms have access to monocular input. Adopting a strategy used by Kolb and Braun (1995. Nature, 377, 336-338), we created drifting textures that were invisible to purely binocular processes. Monocular signals readily conveyed motions defined by local orientation and flicker. However, when left- and right-eye signals were displayed simultaneously, only flicker motion was visible. We conclude that motion-from texture mechanisms do not have access to monocular input. Further evidence suggests that motion from texture involves attentional tracking. PMID- 10367050 TI - Endothelial nitric oxide synthase (eNOS) is localized to Muller cells in all vertebrate retinas. AB - The distribution of endothelial nitric oxide synthase immunoreactivity (eNOS-IR) was investigated in the retinas of all phylogenetic vertebrate classes by using a monoclonal eNOS antibody. Confocal light microscopy showed immunoreactive labeling in Muller cells of fish, frog, salamander, turtle, chicken, rat, ground squirrel, and monkey retina. In vascularized retinas (rat, monkey), astrocytes and some blood vessels were also stained. Furthermore, eNOS-IR was localized to axon terminals of turtle and fish horizontal cells. These observations are the first to show the presence of eNOS-IR in Muller glia and horizontal cell structures of the vertebrate retina. PMID- 10367051 TI - Focal attention in visual search. AB - Visual search operates in different modes assumed to reflect serial and parallel processing. The basis of this distinction is not yet clear. It is often assumed that serial search involves sequential shifts of focal attention across a scene and that no such shifts occur in parallel search. Direct measurements of attention effects during search show that the focus of attention moves to the target (and away from non-targets) both in serial and parallel search. This suggests that the two search modes do not differ in their attentional load but perhaps in the way in which focal attention is directed to the target. PMID- 10367052 TI - A comment on 'Photoreceptor function in unilateral amblyopia'. PMID- 10367053 TI - Stereopsis from contrast envelopes. AB - We report two experiments concerning the site of the principal nonlinearity in second-order stereopsis. The first exploits the asymmetry in perceiving transparency with second-order stimuli found by Langley et al. (1998) (Proceedings of the Royal Society of London B, 265, 1837-1845) i.e. the product of a positive-valued contrast envelope and a mean-zero carrier grating can be seen transparently only when the disparities are consistent with the envelope appearing in front of the carrier. We measured the energy at the envelope frequencies that must be added in order to negate this asymmetry. We report that this amplitude can be predicted from the envelope sidebands and not from the magnitude of compressive pre-cortical nonlinearities measured by other researchers. In the second experiment, contrast threshold elevations were measured for the discrimination of envelope disparities following adaptation to sinusoidal gratings. It is reported that perception of the envelope's depth was affected most when the adapting grating was similar (in orientation and frequency) to the carrier, rather than to the contrast envelope. These results suggest that the principal nonlinearity in second-order stereopsis is cortical, occurring after orientation- and frequency-selective linear filtering. PMID- 10367054 TI - Development of spatial and temporal vision during childhood. AB - Using the method of limits, we measured the development of spatial and temporal vision beginning at 4 years of age. Participants were adults, and children aged 4, 5, 6, and 7 years (n = 24 per age). Spatial vision was assessed with vertical sine-wave gratings, and temporal vision was assessed with an unpatterned luminance field sinusoidally modulated over time. Under these testing conditions, spatial contrast sensitivity at every frequency increased by at least 0.5 log units between 4 and 7 years of age, at which point it was adult-like. Grating acuity reached adult values at 6 years of age. Temporal vision was more mature: at 4 years of age temporal contrast sensitivity at higher temporal frequencies (20 and 30 Hz) and critical flicker fusion frequency were already adult-like. Sensitivity at lower temporal frequencies (5 and 10 Hz) increased by 0.25 log units after the age of 4 to reach adult levels at age 7. The results suggest that temporal vision matures more rapidly than spatial vision during childhood. Thus, spatial and temporal vision are likely mediated by different underlying neural mechanisms that mature at different rates. PMID- 10367055 TI - First and second-order contributions to surface interpolation. AB - Comparisons of 1st- and 2nd-order stereopsis have typically employed isolated, or local, narrow-band targets. While these experiments have revealed a great deal about the distinction between these two types of processing, such stimuli are rare in the natural environment. Instead, local disparity signals are more likely to be part of extended surfaces that very smoothly in depth. The aim of the experiments presented here is to determine the relative contribution of 1st- and 2nd-order stereopsis to the perception of depth-modulated surfaces. Stereothresholds were measured under a range of conditions designed to isolate either 1st- or 2nd-order processing. The results demonstrate that while 2nd-order stereopsis provides local depth estimates for individual texture elements, 1st order processing is essential to the global interpolation of those estimates across surfaces. PMID- 10367056 TI - Vernier judgments in the absence of regular shape information. AB - Vernier acuity is a form of hyperacuity in which the threshold offset between a test object and a reference object is smaller than the size of a foveal cone. Because the test and the reference objects usually have regular shapes (e.g. rectangular, triangular or circular), relatively few studies have addressed the role of shape information in determining hyperacuity thresholds. In this study, we investigated the effect of shape information on hyperacuity performance using targets of irregular shape with different skew and symmetry properties. Vernier thresholds smaller than 10 arc-sec were obtained for closely spaced asymmetric irregular-shape targets. Thresholds for dots and asymmetric irregular shapes increased with increase in center-to-center gap between the targets. Unlike dots, the thresholds for asymmetric irregular shapes also increased with target area. Although the thresholds for asymmetric irregular shapes were higher than those for dots, thresholds for symmetric irregular shapes were similar. Target skew below a certain level had a negligible effect on Vernier thresholds for asymmetric shapes. Our results suggest the existence of feature-independent neural circuitry that can support hyperacuity thresholds and are consistent with the use of the centroid as a primitive for relative localization. PMID- 10367058 TI - Wondering about the wandering cyclopean eye. AB - Arguments against recent claims (Erkelens, Muijs & van Ee (1996). Vision Research, 36, 2141-2147; Mansfield & Legge (1996). Vision Research, 36, 27-41. (1997), Vision Research, 37, 1610-1613) that the position of the cyclopean eye is stimulus specific are presented. Critical to these arguments are the differences between relative and absolute visual direction tasks (Howard (1982). Human visual orientation. Wiley, New York; Ono & Mapp (1995). A restatement and modification of Wells-Hering's laws of visual direction. Perception, 24, 237-252), and between physical and perceptual descriptions of visual direction (Ono, Ohtsuka & Lillakas (1998). Proceedings of the international workshop on advances in research on visual cognition, Tsukuba, Japan (pp. 125-136); Ono & Lillakas (1997). Proceedings of the fourth international display workshop, Nagoya, Japan (pp. 831 834)). PMID- 10367057 TI - The Poggendorff illusion: a bias in the estimation of the orientation of virtual lines by second-stage filters. AB - The veridical perception of collinearity between two separated lines is distorted by two parallel lines in the space between them (the Poggendorff illusion). This paper tests the conjecture that the perception of collinearity of separated lines is based on a two-stage mechanism. The first stage encodes the orientation of the virtual line between the proximal terminators of the target lines. The second stage compares this virtual orientation with the orientation of the target lines themselves. Errors can and do arise from either process. Two parallel lines, abutting against the target lines, cause the classical Poggendorff misalignment bias. The magnitude of the bias is increased by Gaussian blur, as is a version of the Poggendorff figure containing only acute angles. In the obtuse-angle figure, on the other hand, blur decreases the misalignment bias. We argue that the acute- and obtuse-angle biases depend upon different mechanisms, and that the obtuse angle effect is more related to the obtuse-angle version of the Muller-Lyer illusion, which is also decreased by blur. If observers attempt to match the orientation of the virtual line between the two line intersections in the Poggendorff figure they make an error in the same direction as the Poggendorff bias. The orientation of the target lines in the figure, however, is veridically matched to a Gabor-patch probe, unless the target lines are very short, in which case the error is in the same direction as the Poggendorff bias. A small bend in the target lines where they abut the parallels increases the Poggendorff bias if it makes the line more orthogonal to the parallel, but has little effect in the opposite direction. The Poggendorff bias is unlikely to depend upon biases in first-stage linear filters because (a) it still exists in figures composed of short, luminance-balanced lines which are defined by contrast only; and (b) it also exists if the parallels are replaced by grating patches with the same mean luminance as the background. The orientation of the grating in the latter case affects the magnitude of the bias, but even an orientation which should reverse the Poggendorff bias by the mechanism of cross-orientation inhibition fails to do so. The Poggendorff bias is a complex effect arising from several sources. Blurring in second-stage filters with large receptive fields can explain many aspects of the phenomenon. PMID- 10367059 TI - Modelling spatial contrast sensitivity functions for chromatic and luminance modulated gratings. AB - We extended our detection model of achromatic spatial vision (Rovamo, J., Mustonen, J., & Nasanen, R. (1994a). Modelling contrast sensitivity as a function of retinal illuminance and grating area. Vision Research, 34, 1301-1314) to colour vision by taking into account the fact that due to the spatio-chromatic opponency of retinal ganglion cells and dorsal lateral geniculate nucleus (dLGN) neurons, equiluminous chromatic gratings are not affected by precortical lateral inhibition. We then tested the extended model by using Mullen's experimental data (Mullen, K. J. (1985). The contrast sensitivity of human color vision to red green and blue-yellow chromatic gratings. Journal of Physiology, 359, 381-400). The band-pass shape of the spatial contrast sensitivity function for luminance modulated green and yellow gratings transformed to a low-pass shape, resembling the chromatic spatial contrast sensitivity function for red-green and blue-yellow equiluminous gratings, when the effect of precortical lateral inhibition on grating contrast was computationally removed by dividing luminance contrast sensitivities by spatial frequency (i.e. by af, where a = 1 degree). After the removal of this direct effect of lateral inhibition, there still remained a residual shape difference between the spatial contrast sensitivity functions for chromatic and luminance gratings. It was due to indirect reduction of grating visibility by quantal noise high-pass filtered by precortical lateral inhibition. When this indirect effect of quantal noise was also removed, contrast sensitivity for luminance gratings was about twice the sensitivity for chromatic gratings at all spatial frequencies. This was evidently due to the fact that the chromatic contrast of the equiluminous grating at the opponent stage (Cole, G. R., Hine, T. & McIihagga, W. (1993). Detection mechanisms in L-, M-, and S-cone contrast space. Journal of the Optical Society of America A, 10, 38-51) was about half of the luminance contrast of either of its chromatic component. Thus, if the contrast of the equiluminous chromatic grating were not expressed as the Michelson contrast of one chromatic component grating against its own background (Mullen, K. J. (1985). The contrast sensitivity of human color vision to red green and blue-yellow chromatic gratings. Journal of Physiology, 359, 381-400) but as chromatic contrast at the opponent stage, contrast sensitivity would be the same for chromatic and luminance gratings. PMID- 10367060 TI - Behavioral evidence for visual perception of 3-dimensional surface structures in monkeys. AB - Human subjects perceive two crossing bars, one in front of the other, when shown a cross with disparity added to its horizontal limbs, and they also perceive neon color spreading when shown a stereoscopic Redies-Spillmann figure. It has thus been hypothesized that the human visual system follows the principle of generic image sampling in reconstructing 3-dimensional (3-D) surface structures. Here we examine whether monkeys also perceive these surface structures. The results indicate that monkeys, like humans, perceive two crossing bars and neon-color spreading and suggest that the principle of generic image sampling may also be applied to visual perception in monkeys. PMID- 10367061 TI - How does noise influence the estimation of speed? AB - Local motion signals have to be combined in space and time, to yield a coherent motion percept as it is involved in a variety of visual tasks. This combination necessarily means to trade-off between loosing spatio-temporal resolution by pooling local signals and maintaining perceptually significant segmentation between them. When signals are pooled to detect the presence of coherent motion in large amounts of random noise, the question raised is how the noise affects the perceived quality, in particular speed, of the coherent motion. Is there an analogy to the well-known reduction in the perceived speed of moving gratings at low contrast? Using a two-interval forced-choice procedure, we have investigated the assessment of speed in random-dot kinematograms containing different proportions of noise. Under the conditions investigated, there is no strong reduction of perceived speed with increasing noise, as long as coherence levels remain well above the thresholds for directional judgements. This basic result, which could suggest considerable but not perfect segregation of signal and noise motion components in the pooling process leading to speed estimation, is discussed in relation to a model that is designed to decode speed from a population of elementary motion detectors (EMDs) of the correlation type. A strategy to estimate speed from a set of EMDs with a variety of spatio-temporal tuning does not only provide a velocity predictor unambiguous with the spatial structure of the stimulus, but also is largely independent of noise. PMID- 10367062 TI - Psychophysical observations concerned with a foveal lesion (macular hole). AB - A not uncommon occurrence in elderly people is the development of a 'macular hole' but very few psychophysical observations have been made in such cases. I describe here some distortions of image experienced when I view objects with my right eye which has a macular hole. Objects are distorted by being shrunk towards the fovea. Thus, a disc retains its shape but becomes smaller whereas lines are broken or bent. By analogy with animal experiments it is suggested that the perceptual changes are due to physiological changes in the visual cortex. PMID- 10367063 TI - Temporal resolution deficits in the visual fields of MS patients. AB - We assessed the relationship between temporal resolution and MS-induced neuropathy. A diagnostic strategy comprising assessments of temporal resolution at 16 points in the extra-foveal visual field up to 12 degrees from the fovea was first compared with foveal temporal resolution and with a standard VEP procedure in the same MS patients. At the group level, foveal temporal resolution was less sensitive to demyelination than the 16-point diagnostic strategy, the detection rate of which was comparable to that of the VEP procedure. Cross-sensitivity of the VEP and the 16-point diagnostic procedure was low. Subsequently, the average severity of MS-induced temporal resolution deficits was studied at three retinal loci of the same size but different eccentricities. Foveal deficits were not significantly greater than more peripheral deficits within the central 12 degrees. PMID- 10367064 TI - [Chronic laryngitis in children: the role of gastroesophageal reflux]. AB - Gastro-oesophageal reflux (GOR) is associated with a number of inflammatory ENT disorders in the adult and is correlated with recurrent croup in the child. AIM: To estimate the frequency of GOR in a population of children consulting for chronic laryngotracheal symptoms. METHOD: The study included 17 children, aged between 2 and 14 years (mean: 7 years) all of whom suffered from dysphonia or a chronic cough. After a clinical ENT examination, each child had a fibreoptic laryngoscopy and a long duration pH-study lasting between 18 and 24 hours. RESULTS: Pathological GOR was discovered in 10 children, i.e. 59%. Overall the number of refluxes per study varied from 6 to 816 (mean 156). The vast majority of these refluxes occurred when the child was awake. CONCLUSION: In our series of children with chronic laryngotracheal disorders, at least 59% were shown to suffer from pathological GOR. PMID- 10367065 TI - [Variations and incidents encountered during stapes surgery for otosclerosis]. AB - The aim of this study was to report peroperative variations and incidents encountered during stapes surgery for otosclerosis, from a retrospective series of 293 primary stapedectomies performed in the ENT department of Fondation Adolphe de Rothschild, from january 1990 to december 1996. Pitfalls related to anomalous states or pathological conditions were observed in 81 cases (27.6%) and were classified as follow: narrow oval window niche (12.3%), floating footplate (5.8%), obliterative otosclerosis (4.7%), ossicular chain problems (3.4%), loss of perilymph and/or intravestibular bleeding (2.4%), fixed malleus (1%). Hearing results following surgery are presented. Because of the relatively high incidence of these unforeseen events, the otologic surgeon must be well trained and experienced to be able to use all reasonable stapedectomy or stapedotomy techniques. PMID- 10367066 TI - [The use of calvarial bone in nasal reconstruction]. AB - Twenty-seven nasal reconstructions with parietal bone graft performed between January 1989 and January 1995 were analyzed retrospectively. No complications involving parietal bone were observed. After a 2-year follow-up there have been no cases of shifting, extrusion or resorption of the rhinoplasty. Our experience shows that calvarial bone is an ideal material for nasal reconstruction bone grafts. PMID- 10367067 TI - [Otosclerosis surgery: a series of 227 cases. Introduction of CO2 laser]. AB - OBJECTIVES: To evaluate the effects of the size of the footplate opening on the hearing results in surgical treatment of otosclerosis and the use of CO2 laser in this indication. PATIENTS AND METHODS: 190 patients with otosclerosis underwent 227 procedures between 1986 and 1995. Hearing results and symptoms were analyzed to compare the different procedures: 140 stapedectomies, 87 Fisch's stapedotomies, 35 of them with manual perforator, 52 of them with CO2 laser. RESULTS: Air/bone gap closure within 10 dB was obtained in 87, 92 and 97 percent of stapedectomies and in 80, 84 et 90 percent of stapedotomies after 3 months, 1 and 3 years (NS). Bone conduction was improved in 81, 80 et 63 percent of stapedectomies and 87, 97, 60 percent of stapedotomies after the same time (NS). Air/bone gap closure within 10 dB was obtained in 75 and 80 percent of manual perforator stapedotomies and in 84 and 88 percent of CO2 laser stapedotomies after 3 months, and 1 year (NS). Bone conduction was improved in 78 and 96 percent of manual perforator stapedotomies and 95 and 100 percent of CO2 laser stapedotomies after the same time (NS). No facial palsy or prolonged vertigo occurred. There was one case of anucusis following a stapedectomy. CONCLUSION: Although both stapedectomy and stapedotomy produced equivalent air/bone gap closure, reduction of inner ear trauma was noted with Fisch's stapedotomy. CO2 laser technique is a safe procedure, which optimizes the Fisch's stapedotomy. PMID- 10367068 TI - [Bilateral rhinolithiasis by transseptal permeation: a case report]. AB - We report a case of bilateral rhinolithiasis due to transseptal destruction. The lesion developed over several years leading to bilateral nasal obstruction and recurrent purulent fetid nasal discharge. At rhinoscopy, the lesion was located in the posterior part of the nasal fossae. CT-scan showed total destruction of the bony septum and a transversal rhinolith anterior to the choanal orifices. Extraction was performed under local anesthesia. Follow-up endoscopy confirmed transseptal permeation. PMID- 10367069 TI - [Contribution of vastarel 20 mg (trimetazidine) in the treatment of vertigo: synopsis of clinical trials]. PMID- 10367070 TI - [Congenital cardiopathies in adult age. A growing problem. A new "subspecialty"?]. PMID- 10367071 TI - [Congenital long QT syndrome. The value of genetics in prognostic evaluation]. AB - The congenital long QT syndrome (QTL) is a heterogenic clinical and genetic entity characterised by prolongation of the QT interval which may be complicated by syncope and sudden death. Four genes have been identified for the cardiac potassium (KCNQ1, HERG and KCNE1) and sodium (SCN5A). The aim of this study was to assess the prognosis of the disease by the site of mutation identified on the morbid gene. Thirty-two genotyped families participated to this study. Each subject gave a clinical history, an ECG and a search for genetic mutation. Eighteen mutations in the transmembrane domains of KCNQ1 were identified in 25 families and 2 mutations in the C-terminal part were found in 4 families. The phenotype was less severe in C-terminal part mutations: less syncopes and sudden deaths (22 vs 55%, p < 0.001) and a shorter QTc (458 +/- 31 ms vs 479 +/- 31 ms, p = 0.0003). Three mutations were detected in the C-terminal part of HERG in 3 different families. Their phenotype was less severe with syncoped related to hypokalemia. The authors also report the case of a family in which two subjects who were the most severely affected had two mutations, one in HERG and the other in KCNQ1. This study confirms the value of a genetic research in assessing the severity of the congenital long QT syndrome. PMID- 10367072 TI - [Percutaneous closure of ostium secundum atrial septal defects using the Amplatz device. Preliminary study on 29 patients]. AB - The aim of the study was to assess the results of percutaneous closure of ostium secundum atrial septal defects (ASD) by the Amplatz device and to compare them with those obtained by other methods. Over a one year period, closure of ASD was performed in 28 patients aged 7 to 66 years (mean of 32 years) with exclusive left-to-right shunts. The selection of patients was made by transthoracic echocardiography. Cardiac catheterisation was performed to measure the stretched diameter of the defect. After patient or parental consent, the procedure was performed under general anaesthesia. The implantation was performed under transoesophageal echocardiographic control. The diameter of the Amplatz device corresponded to the stretched diameter of the ASD. The patients were discharged from hospital 48 hours later under platelet anti-aggregant therapy for 6 months. Clinical, electrocardiographic, radiological and echocardiographic examinations were performed at 1 month, 3 months and 1 year after implantation. The stretched diameter was 12-27 mm (mean 22 mm). The device was withdrawn in one case because of a double ASD and, in another patient, failure was due to embolisation of the obturator to the pulmonary artery before its implantation. The closure was complete in 21 of the 26 cases immediately after the procedure; at one month, two residual shunts were observed but they had disappeared at 3 months and at 1 year. Out of 46 ASD closed by the Sideris button prosthesis, occlusion was total in 29 cases and partial in 17 cases. The authors conclude that the Amplatz device is an effective prosthesis for closure of ostium secundum ASD. With strict selection procedures, the results are excellent. PMID- 10367073 TI - [Surgical evaluation of transthoracic tridimensional echocardiography in the anatomic study of atrial septal defect]. AB - The closure of atrial septal defects by interventional catheterisation requires an accurate assessment of their morphology and anatomical relationships. This study evaluated transthoracic three-dimensional echocardiography for the selection of atrial septal defects accessible to an occlusive prosthesis. The transthoracic three-dimensional echocardiographic measurements of 17 patients (4 to 55 years) with ostium secundum atrial septal defects were compared with those of the surgeon in a prospective study. The maximal diameters of the defect, the height of the interatrial septum, the distances to the superior vena cava (postero-superior border) and inferior vena cava (postero-inferior border), to the coronary sinus and the tricuspid valve were measured as a reconstruction of the interatrial septum seen from the right atrium. The aortic border was measured from a three-dimensional view from the left atrium. Thirteen of the 17 investigations (76%) were exploitable. The diameters of the defect varied during the cardiac cycle (p = 0.0002). Ther correlations between the surgical and echocardiographic measurements varied from 0.82 for the maximal diameter to 0.6 for the postero-inferior limits. Three-dimensional echocardiography is capable of detecting all the contra-indications of an occlusive prosthesis: 2 inadequate postero-inferior and 1 inadequate aortic borders, 9 maximal diameters which were too large, 3 insufficiently high atrial septa, 1 double atrial septal defect. The coronary sinus was only visualised in 1 case. Transthoracic three-dimensional echocardiography is a non-invasive technique capable of improving the selection of atrial septal defects for interventional closure. The transoesophageal approach should be reserved for candidates selected by the transthoracic investigation for the detection of small structures (coronary sinus) and when the transthoracic window is poor. PMID- 10367074 TI - [A new anatomical approach to congenital valve diseases by three-dimensional echocardiography]. AB - Congenital valvular heart disease in childhood is often complex. Conventional echocardiography provides two-dimensional views which require mental reconstruction for three-dimensional assessment. This problem may be solved by the use of three-dimensional (3D) echocardiography which obtains images of valves comparable to those seen at surgery. This was confirmed by 4 cases of congenital valvular heart disease studied by 3D echocardiography: stenotic bicuspid aortic valve disease, parachute mitral valve and two cases of mitral regurgitation in patients with atrioventricular canal. The 3D views of the aortic valve showed the commissural opening after percutaneous balloon valvuloplasty of the bicuspid valve. The surface of the aortic orifice and the surface of the two mitral leaflets were measured from 3D reconstructions. The longitudinal 3D view analysed the extension of the single obstructive mitral papillary muscle of the parachute malformation. The 3D ventricular views allowed assessment of the extension of the cleft and the surface of the three mitral leaflets of the 2 cases of atrioventricular canal. In one of these cases, the results of surgical valvuloplasty were evaluated after 3D reconstruction of the valve. 3D echocardiography is not only a diagnostic tool for congenital heart disease but also a very useful complementary investigation for accurate evaluation of congenital valvular lesions to optimise possible valve repair. PMID- 10367075 TI - [Treatment of obstruction of prosthetic conduct by percutaneous implantation of stents]. AB - Prosthetic conduits, with or without biological valves, are often inserted in surgical procedures to correct or palliate cardiac malformations. The principal problem is degeneration which causes variable degrees of obstruction requiring reoperation for their replacement. The aims of this study were to assess the feasibility, safety and efficacy of a non-surgical method of treating these obstructive prostheses by dilatation-implantation of a metallic vascular endoprosthesis (stenting). Thirteen patients were treated (age range 7.7 to 36 years; mean: 15 years). Eight had pulmonary atresia with a ventricular septal defect corrected by a valved conduit from the right ventricle to the pulmonary artery which became obstructive nearly 10 years later: the implantation of the stent reduced the transconduit pressure gradient in all cases except one who had not undergone closure of the septal defect in which the cyanosis was improved. There are two cases of obstruction of a modified Blalock anastomosis in which the stent revascularised the shunt with improvement in cyanosis. In the final 3 cases, the whole Fontan procedure was compromised by obstruction of a conduit incorporated in the system, and which dilatation with stenting considerably improved. The efficacy of the procedure was constant with no complications other than rupture of the balloon in 3 cases. The good results were maintained for an average of 7.3 months (range 1 to 25 months), but it was necessary to redilate one restenosed stent after 8 months. Dilatation followed by stenting in obstructive cardiovascular prostheses is a simple, safe and effective alternative to surgical reoperation. PMID- 10367076 TI - [Biventricular repair in neonates with hypoplasia of the left ventricle]. AB - BACKGROUND: Whether to perform uni or biventricular repair in ducto dependent neonates with hypoplastic but morphologically normal left ventricle and multi level left ventricle obstructions (hypoplastic left heart syndrome class III) remains unanswered. Echocardiographic criteria have been proposed for surgical decision. HYPOTHESIS: Increased afterload and multi level left ventricle obstruction is constant. We assumed that restoration of normal loading conditions by relief of left ventricle obstructions promotes its growth, provided that part of the cardiac output was pre operatively supported by the left ventricle, whatever the echocardiographic indices. METHODS: Twenty one ducto dependent neonates presented with this anomaly. All had aortic coarctation associated to multi level left ventricle obstruction. Pre operative echocardiographic assessment showed: mean end diastolic left ventricular volume of 13.3 +/- 3.5 mL/m2 and mean Rhodes score of -1.43 +/- 0.9. Surgery consisted in relief of left ventricle outflow tract obstruction by coarctation repair in 21 associated to atrial septal defect closure in 2, aortic commissurotomy in 1 and ascending aorta enlargement in 1. RESULTS: There were 3 early and 3 late deaths. There was no predictive risk factor for failure. Growth of the left heart was demonstrated in most patients. At hospital discharge the end diastolic left ventricular volume was 19.4 +/- 3.12 mL/m2 (p = 0.0001) and the Rhodes score was -0.38 +/- 1.01 (p = 0.0003). Actuarial survival and freedom from reoperation rates at 5 years were: 68.5% and 40.75%, respectively. CONCLUSION: Biventricular repair can be proposed to ducto dependent neonates with hypoplastic but morphologically normal left ventricle provided that all anatomical causes of left ventricle obstruction can be relieved. Secondary growth of the left heart then occurs, however the reoperation rate is not low. PMID- 10367077 TI - [Immediate and mid-term results of surgery for aortic coarctation in children under 6 months of age]. AB - The aim of this study was to assess the impact of medical and surgical advances on the results of surgery of coarctation of the aorta without major associated cardiac malformations in children aged under 6 months, operated by the surgical team of Tours over a period of 17 years. A retrospective analysis of patients' files allowed inclusion of 75 children: 34 in group I (1980-1988) and 41 in group II (1989-1996). The age at diagnosis, mode of presentation, clinical and echocardiographic parameters, initially and at surgery, were comparable in the two groups. In group II, the surgical procedure of choice changed to the Crafoord procedure (p = 0.0001), the peroperative haemodynamic complications were less common (p = 0.04), patients were operated sooner after diagnosis (p = 0.002) with a higher number of neonates (p = 0.04). There were two early deaths in group I and two late deaths in group II. Six children (8.2%) had recurrence of coarctation. This was more common in children operated before 1 month of age (p = 0.0001) and in cases of hypoplasia of the aortic arch (p < 0.01). The risk of recoarctation in neonates was lower in group II than in group I (p < 0.02). At medium term, the coarctation was considered well repaired in 93.7% and 92.6% of patients in group I and II respectively. However, hypertension on exercise was observed in 8 of the 19 children studied. Finally, secondary surgical or instrumental procedures were required in 9 children to treat associated cardiovascular abnormalities. The authors conclude that in recent years, aortic coarctation was repaired earlier, mainly by a modified Crafoord procedure, with fewer haemodynamic complications, without early mortality. The risk of recoarctation remains low but persistent in neonates. At medium term, hypertension on exercise is a common problem even in children with excellent repairs. PMID- 10367078 TI - [The Ross procedure in the acute phase of infectious endocarditis in childhood]. AB - The Ross procedure of aortic valve replacement with a pulmonary autograft has several advantages in childhood over mechanical prostheses or homografts, especially in infectious endocarditis requiring early surgery. Between January 1997 and July 1998, 3 children with no known previous cardiac disease, aged 14 months, 10 and 11 years, had aortic valve infectious endocarditis. The causal organism was not identified in 1 case and the other two were due to staphylococcus aureus and corynebacterium diphteriae. All children had severe, rapidly progressive aortic regurgitation complicated by pulmonary oedema in the baby and systemic emboli in the two older children. Surgery was performed within 9 days, 1.5 month and 2 months after the onset of the disease. The postoperative course was uncomplicated in the 3 cases. Postoperative Doppler echocardiography showed absence of autograft dysfunction or stenosis, with the presence of pulmonary regurgitation in 1 case. Pulmonary autograft has the advantages of not requiring anticoagulation, of allowing growth of the aortic ring, of not being limited by the age of the patient and of having a low risk of degeneration and infectious endocarditis. Therefore, it seems particularly indicated for cases of complicated infectious endocarditis requiring early aortic valve replacement. The early (4.8%) and late (4.3%) mortality rates were comparable to those of other techniques and are lower than those associated with valve replacement with mechanical prostheses in cases of endocarditis (8.5% versus 40%). The secondary morbidity is 18.8% with dysfunction of the autograft and/or stenosis of the pulmonary homograft. Despite a limited follow-up, aortic valve replacement by a pulmonary homograft seems better than aortic valve replacement with a homograft or mechanical prosthesis, especially in cases of complicated infectious endocarditis requiring surgery in the acute phase. Further studies are required to confirm these encouraging results. PMID- 10367079 TI - [Evolution of ventricular septal defects. Relation to echocardiographic anatomy]. AB - The aim of this study was to analyse the outcome of membranous ventricular septal defects (VSD) with respect to the echocardiographic data obtained during the first year of life. This retrospective series included patients born between January 1st 1986 and December 31st 1995, in the Indre et Loire department, with membranous ventricular septal defects alone or associated with minor abnormalities. The initial echocardiography, an echocardiography performed 2 to 6 months later, one a year later and the final echocardiography were compared. Three groups of VSD were constituted according to their diameter: group I (< or = 3 mm), group II (3-6 mm), and group III (> 6 mm). Depending on the outcome, the patients were classified as spontaneous closure (group A), surgical closure (group B) or persistent VSD (group C). The population comprised 84 children. There were 6 spontaneous deaths, three of which were unexplained, and 7 children were lost to follow-up. After the initial echocardiography, the VSD were classified as group I (38%), group II (26.2%) and group III (35.7%). After the second echocardiogram, 24 VSD changed group (31.5%), by increase (N = 10) or decrease (N = 14) in diameter. Aneurysms of the membranous septum were observed during the first two echocardiographies in 31.2% and 79.3% of VSDs of group I, 31.8% and 70% of VSDs of group II and 6.6% and 3.3% of VSDs of group III (p < 0.01). The average follow-up was 3.1 years (range 1 month-10 years). In group A (N = 22), the mean age of closure of the VSD was 26 months (3 months-7 years). In group B (N = 28), surgery was undertaken at an average age of 10 months (range 3 months-5 years). In group C (N = 21), the VSDs were classified as group I (N = 19) or group II (N = 2) at the last echocardiography. The frequency of aneurysms of the membranous septum in groups A, B and C were respectively 100%, 7.1% and 66.6% (p < 0.01). At the second echocardiographic examination, a significant relationship (p < 0.001) was observed between the diameter of the VSDs and their outcome. The VSDs of group A were associated with aneurysms of the membranous septum more often than those of group C (p < 0.005). The authors conclude that surgery is required in about one third of membranous VSD. At medium term, the others either close spontaneously or become smaller in comparable numbers. The outcome is directly related to the diameter of the VSD and the development of an aneurysm of the membranous septum. During the first 6 months, the dimensions of membranous VSDs change in about 30% of cases with an increase in frequency of aneurysms of the membranous septum. PMID- 10367080 TI - [Role of right-to-left atrial shunt in exercise tolerance of patients with Ebstein anomaly]. AB - The aim of this study was to assess cardiorespiratory tolerance to exercise in children with non-operated, paucisymptomatic and untreated froms of Ebstein's anomaly. The authors undertook a prospective study in 11 children, mean age 9.6 years, who had lung function tests, cardiorespiratory exercise stress tests (bicycle ergometry N = 8, treadmill N = 3) and contrast echocardiography. All parameters of spirometry were normal. Contrast echocardiography showed a right-to left interatrial shunt in 7 children (group 1) whereas the remaining 4 children had no shunt (group 2). The resting oxygen saturation was 97.4 +/- 2%, with no difference between the two groups. On the other hand, oxygen saturation at peak VO2 (VO2 max) was 90 +/- 9.5%, significantly lower in group 1 than in group 2 (85.7 +/- 2.2% vs 98.2 +/- 1.2%; p = 0.03). In group 1, the VO2 max was correlated to oxygen saturation (r = 0.98; p < 0.001, N = 6). The oxygen desaturation was correlated with presence of a right-to-left interatrial shunt (p = 0.01). The reduced exercise tolerance of non-operated, paucisymptomatic children with Ebstein's anomaly is due to a right-to-left interatrial shunt. In patients with poor exercise tolerance, contrast echocardiography is advised for the detection of these atrial shunts. PMID- 10367081 TI - [Prenatal diagnosis of transposition of great vessels reduces neonatal morbidity and mortality]. AB - Transposition of the great arteries (TGA) is a common malformation which sometimes has a dramatic presentation at birth but which is completely curable with early and appropriate initial management. Antenatal diagnosis of this condition may change the neonatal prognosis. The authors compared morbidity and mortality in the pre- and postoperative periods of 68 neonates with an antenatal diagnosis of TGA (foetal diagnosis) with that of 250 neonates in whom the diagnosis was made after birth (neonatal diagnosis). The delay before admission to the department was 2 +/- 2.8 hours in the foetal group and 73 +/- 210 hours in the neonatal group (p < 0.01). Severe haemodynamic distress (metabolic acidosis, multi-organ failure) were more common in the neonatal group (p < 0.01). Management on admission was identical in the two groups (p > 0.05). The preoperative mortality was 15/250 in the neonatal group (6%, 95% CI = 3-9%) compared with 0/68 in the foetal group (p < 0.05). The postoperative morbidity was comparable in the two groups (25/235 and 6/68) but the hospital stay was longer in the neonatal group (30 +/- 17 versus 24 +/- 11 days, p < 0.01). Finally, postoperative mortality was significantly higher in the neonatal group (20/235 compared with 0/68, p < 0.01) although the risk factors of death at arterial switch surgery were identical in the two groups. Therefore, antenatal diagnosis of TGA reduces neonatal morbidity and mortality in this condition. Antenatal diagnosis must be developed by the education of obstetricians. The transfer of mothers with a foetus affected by TGA to centres capable of assuming the initial management, sometimes during labour, is essential. PMID- 10367082 TI - [Right postero-lateral thoracotomy for open heart surgery in infants and children. Indications and results]. AB - In order to avoid the aesthetic prejudice of median sternotomy in young children undergoing open heart surgery for isolated congenital heart disease, a right posterolateral thoracotomy was performed in 146 children aged 5 months to 14 years. The large majority (140/146, 96%) were atrial septal defects: 130 ostium secundum, 5 sinus venosus, 1 low septal defect and 4 ostium primum (partial endocardial cushion defect). Six children had isolated perimembranous ventricular septal defects. One patient died of a probable lesional pulmonary oedema. Minor complications were observed in 15 cases and one had to be reoperated for a residual shunt. With an average follow-up of 2.6 years, all survivors are asymptomatic. The scar is normal, without cheloid or thoracic deformation, invisible to the patient when he looks in a mirror. The use of this approach requires a very accurate anatomical diagnosis, especially with regards to systemic and pulmonary drainage. The presence of a left superior vena cava draining into the coronary sinus is a contra-indication. The right posterolateral thoracotomy is now the approach of choice in its standard indication, the repair of ostium secundum atrial septal defects with large shunts, in young girls under 10 years of age. PMID- 10367083 TI - [Hemangioma and superficial arteriovenous malformations]. AB - Haemangiomas are different from true superficial vascular malformations. The haemangiomas, mainly affecting the newborn and small babies, will, after a phase of progression, sometimes regress completely. Therapeutic abstention is the rule except in high risk angiomas when steroid therapy may be effective. Visceral involvement poses problems. Superficial vascular malformations, on the other hand, arise at all ages and may affect any blood vessel. Each type has a specific clinical presentation, complementary investigations and appropriate treatment. Some are slowly progressive, for example capillary, venous and lymphatic malformations. Others are haemodynamically active, such as the arteriovenous malformations. Capillary malformations are flat angiomas with aesthetic consequences, apart from the Sturge-Weber-Krabbe syndrome. Cold, blue venous malformations confirmed by ultrasonography and magnetic resonance imaging, when necessary, require treatment adapted to their site and size: compression, embolisation, surgery or abstention. Lymphatic malformations may be cystic or tissular: the cystic lymphangioma, a soft swelling of often healthy skin, with compartments separated by septa on ultrasound scan, is usually treated by ethibloc embolisation. Arteriovenous malformations, warm and pulsatile, demonstrated at arteriography, may progress rapidly and treatment by surgery or embolisation, when necessary, has to be complete. Finally, there are complex vascular malformations which pose very difficult problems of management. PMID- 10367084 TI - [Subtotal thrombosis of the juxta-ductal thoracic aorta in a premature 36-week old infant. Alteplase therapy]. AB - The authors report the case of a neonate, a premature 36 weeks gestation male child who presented with a spontaneous thrombosis of the juxtaduetal aorta at 3 days of age. The clinical presentation mimicked that of severe coarctation with cardiocirculatory failure. The diagnosis was ineffective, the introduction of low dose alteplase (plasma activator of human recombinant plasminogen) with continued anticoagulation resulted in near complete lysis of the clot and avoided surgery. Thromboembolic cerebral and renal complications were observed during treatment. However, 6 months after the acute episode, there were no clinical or echographic sequellae. Global renal function remained normal despite mild atrophy of the parenchyma of the left kidney. This combined treatment represents an alternative to high risk surgery. The follow-up of this form of management should be rigorous in view of the potential renal and cerebral complications. PMID- 10367085 TI - [Three different types of atrial septal defects in the same family]. AB - Most of the time atrial septal defects (ASD) are sporadic. Familial forms are characterised by the same type of the ASD, and are frequently associated by other cardiac, osteoarticular (Holt-Oram syndrome) or atrioventricular conductions abnormalities (ostium secundum ASD with prolonged PR, sinus venosus ASD with sinusal bradyarrhythmia). This report describes a family in which the father and his two children present an ASD, each one of a different type (ostium primum ASD, ostium secundum ASD, sinus venosus ASD), without any other associated malformations. To the best of our knowledge, this is the first report of a family in which three different ASD types are present. PMID- 10367086 TI - [Meeting of the Pediatric Cardiology Subcommittee of the French Society of Cardiology]. PMID- 10367087 TI - [Triggering factor in acute myocardial infarction: role of intense physical exercise, anger and sexual activity]. PMID- 10367088 TI - [Study of the relaxation phase of the right ventricle]. AB - The purpose of our study was to investigate the existence or not of an isovolumic relaxation period in the right ventricle in experimental animals with normal pressures in the pulmonary artery. Right and left ventricular pressures, pulmonary and aortic pressures (microtransducers), pulmonary flow, ventricular diameters (sonomicrometer), were recorded at the same time, in 10 sheep anesthetized intravenously with pentobarbital. We obtained "off line" the first ventricular pressures derivative, the ventricular volumes and the pressure-volume loops of both ventricles. The minimum systolic right ventricular volume coincided with 0 pulmonary flow, and both with a diastolic pressure value of 0-5 mmHg in that ventricle. Once the minimum systolic volume was reached, a rapid increase of the right ventricular volume started. The right ventricular pressure-volume loop, unlike the left ventricular one, adopted a non-rectangular shape. The right ventricular ejection period lasted until the beginning of the next filling phase. We concluded that there is no right ventricular isovolumic relaxation period. PMID- 10367090 TI - [Structural and functional changes in the of heart of high-performance (canoeing) athletes]. AB - We studied two groups of healthy subjects: Group I was integrated by 13 high performance sportsmen (10 men and 3 women), devoted to the discipline of the rowing. Group II was integrated by 16 sedentary healthy subjects. All of them were studied with a two-dimensional echocardiogram, in order to study the anatomical and functional characteristics of the heart. Both groups had similar characteristics in regard of total body area, heart rate and blood pressure, the only difference was in age. The ventricular mass and the diastolic volume were greater in athletes in spite of the fact that the dimensions and transverse thicknesses were similar, this imply a longitudinal increase of the heart size. It is possible that this form of ventricular remodeling has functional advantages. On the other hand, it was demonstrated the existence of physiological hypertrophy without disorders in diastolic function. PMID- 10367089 TI - Ebstein's anomaly. Clinical profile in 174 patients. AB - The study population consisted of 148 patients who did not undergo surgical treatment and 26 who were operated, most of them diagnosed after the age of 2, with a follow-up from 6 months to 25.3 years. Patients were divided in three groups of clinical deterioration according to their functional class and cardiothoracic index (CTR) long-term follow-up in 148 nonoperated patients showed significant differences for mortality between groups I and III (p < 0.001), and between groups II and III (p < 0.02). Predictors of death included the association among functional class III or IV CTR > or = 65% with either cyanosis or arrhythmias (p < 0.05). The multivariate analysis showed that clinical deterioration (p < 0.0001), CTR (p < 0.0002) and functional class (p < 0.001), were significant for mortality. Kaplan-Meier analysis showed a survival rate of 81% in the overall patients free from surgical treatment. According to Kaplan Meier analysis, the rate of survival was lower in patients with CTR > or = 65% (63.5%), in patients who had functional class IV (52.5%) and in patients included in group III of clinical deterioration (38.2%). Despite the fact that the association of functional class III or IV plus CTR > or = 65% with either cyanosis or arrhythmias is a good predictor for death, the mortality in patients who had only one of these variables was lower. Patients included in group II of clinical deterioration in stable condition presented long survival with medical treatment. Due to the high mortality rate found in group III, surgical treatment of Ebstein's anomaly must be done before deteriorating into group III. Surgical indication must be done considering the surgical risk of each group according to the experience of the Institution and comparing the rate of surgical mortality with the rate of survival without surgery. PMID- 10367091 TI - [Chagas cardiomyopathy and the autonomic nervous system. Clinical studies]. AB - The autonomic nervous system is abnormal in patients with advanced Chagas'heart disease. Most researchers consider these autonomic abnormalities as primary, specific and irreversible. However, when and why do these abnormalities appear in the natural history of Chagas'disease, is still the subject of intense controversy. Recent morphological and functional studies strongly suggest that, the sympathetic and the parasympathetic abnormalities, are preceded by myocardial damage and left ventricular dysfunction. Consequently, the cardiac parasympathetic abnormalities and the activation of the sympathetic and of the renin-angiotensin-aldosterone systems of chagasic patients are very likely secondary and partially reversible. Therefore, neurohormonal activation, as postulated by the neurogenic theory could contribute to the progression of myocardial damage and left ventricular dysfunction. Additional clinical investigations are necessary to clarify this important issue. PMID- 10367092 TI - [Evaluation of the complications of different types of right ventricular bypass by contrast echocardiography]. AB - We evaluated 54 patients with different right heart by-pass by means of contrast echocardiography with rapid venous injection of shaken saline (3 cc, < 20 k; 6cc, > 20 k), in order to detect venous collateral circulation in partial by-pass, pulmonary arteriovenous fistulae in partial and total by-pass, and right-to-left shunt in total by-pass. Forty three patients had partial by-pass: 29 had a bidirectional cavopulmonary anastomosis with additional pulmonary flow (systemic pulmonary anastomosis and/or restrictive anterograde ventricular flow); 10 had a partial ventricular correction (bidirectional cavopulmonary anastomosis and non restrictive anterograde ventricular flow; two had classical Glenn procedures; two had Kawashima operations (bidirectional cavopulmonary anastomosis and non restrictive anterograde ventricular flow; two had classical Glenn procedures; two had kawashima operations (bidirectional cavopulmonary anastomosis with inferior vena cava interruption), and 11 with total by-pass (10 atriopulmonary anastomosis and 1 with total cavopulmonary anastomosis). The age ranged from 2.5 to 33 years (x = 12.2 years), and the mean postoperative period was 4.3 years. Venous collateral circulation: 32/43 patients (74%) with partial by-pass, specially in those without partial biventricular correction: 29/33 patients (88%) compared to those with partial biventricular correction: 3/10 (30%) p < 0.01. Pulmonary arteriovenous fistulae: 6/43 patients (14%) with partial by-pass; 6/33 (18%) with partial biventricular correction, 0/10 patients with partial biventricular correction, and 0/11 with total by-pass. Right-to-left shunt: 2/11 patients (18%) with total by-pass, all of them with atriopulmonary anastomosis. Contrast echocardiography is an excellent noninvasive method for the initial evaluation of specific dysfunctions of different right heart by-pass. We emphasize the higher frequency of collateral venous circulation in cavopulmonary anastomosis, that would explain the dysfunction with progressive hypoxia in the evolution of these patients. Pulmonary arteriovenous fistulae were detected only in partial by-pass, without partial biventricular correction (p < 0.01). The minor frequency of these fistulae in bidirectional cavopulmonary anastomosis would be due to additional pulmonary flow. PMID- 10367093 TI - [Provocation and measurement of retrograde pulsatile wave velocity in normal subjects and patients with systemic hypertension]. AB - There are not published incruent methods of provocation and/or measurement of the velocity of the reflexive arterial pulse wave. This phenomenon is implicated in the pathogenesis of arterial hypertension. We describe that during compressive sphygmomanometry (CS) done in the forearm, reflexive waves are provoked that are register in the arm with an equipment developed by us, which measures the velocity of the antegrade (APWV) and the provoked retrograde (RPWV) pulse waves. The procedure consist in: 1) detection, capture and digitalization by pneumatic cuffs of oscillopulses of the arm and the forearm, 2) detection of finger photopulse to control the efficacy of CS, 3) measurement of APWV and RPWV by taking the conduction time travel distance of pulse wave between detectors arm forearm and forearm-arm respectively. Thirty normal case (group A, GA) and 37 essential systemic hypertensive patients (group B, GB) were studied. Sixteen cases of GB had left ventricular hypertrophy (LVH). RESULTS: A reflexive wave was provoked in 99% of cases. The APWV (m/s) was 6.0 +/- 0.9 vs 7.5 +/- 1.3, p < 0.001 for GA and GB respectively. The RPWV (m/s) of the same groups were 1.8 +/- 0.3 vs 2.5 +/- 0.7, p < 0.001. The hypertensive cases with LVH had more RPWV than the cases without it (2.7 +/- 0.6 vs 2.3 +/- 0.6, p = 0.07). CONCLUSION: Hypertensive cases vs normals had higher antegrade and retrograde pulse wave velocities possible due to the major degree of arterial rigidity of the former. The method may be used in study of chronic arteriopathy. PMID- 10367094 TI - [Congenital atrioventricular junction tachycardia. Favorable response to anti arrhythmia agents]. AB - The congenital junctional ectopic tachycardia, is an unusual tachyarrhythmia, with early clinic manifestation and poor antiarrhythmic drugs response with a great infant mortality reaching rates of 35%. It deserves a special attention in its timely detection as well as in its appropriate handling with different modalities of pharmacological and nonpharmacological therapy. We reported two cases in which age of presentation of tachyarrhythmia was at three months and whose particularity was the good response to antiarrhythmic drugs; verapamil and later propafenone, used for the acute stages and a combination of propafenone plus propranolol initially for the chronic stage. Due to intolerance it was necessary to change the treatment after a year for sotalol and digital with good response. We review the literature about this topic. PMID- 10367095 TI - Interventional cardiology in congenital heart disease. AB - Interventional procedures for congenital heart disease have evolved dramatically in the last decade. Beginning with pulmonary and aortic valvuloplasty, nowadays, stents and various devices are placed inside the heart and vessels to palliate or correct different congenital defects. The present communication summarizes the experience with interventional cardiology in congenital heart disease of the Instituto Nacional de Cardiologia "Ignacio Chavez" during the last five years. PMID- 10367096 TI - [Quadricuspid aorta diagnosed by transesophageal echocardiography. A case report]. AB - We report a case of a 64 year-old woman who had a quadricuspid aortic valve associated with aortic regurgitation of mild degree diagnosed by transesophageal echocardiography (TEE). The four cuspids were of the same size. This valve anomaly could not be diagnosed by transthoracic echocardiogram (TTE). The TEE is a diagnostic noninvasive procedure that gives the best information in quadricuspid aortic valve malformation. PMID- 10367097 TI - [Participation of major histocompatibility system alleles in the susceptibility to rheumatic fever]. PMID- 10367099 TI - Placebo--efficacy and adverse effects in controlled clinical trials. AB - The therapeutic efficacy of placebo in a series of diseases has long been known. It is less well known, however, that treatment with placebo can also produce significant adverse drug reactions. Therefore, the placebo drug reactions from controlled trials were studied for the first time systematically. METHOD: The efficacy and the safety of placebos were investigated using patient and drug data pooled from randomized, placebo-controlled, multicentre studies in five different groups of indications covering the therapeutic areas of cardiology (nisoldipine), neurology/psychiatry (nimodipine/ipsapirone), metabolism (acarbose) and gastroenterology (hydrotalcite). RESULTS: The efficacy of placebo was clear, and varied not only between the five indication groups but also within them. Whereas placebo, unlike active treatment, produced hardly any improvement in symptoms in patients with severe stroke, it was as effective as active treatment in patients with mild neurological deficits, producing an improvement of about 50%. In patients with angina pectoris, placebo produced an increase in exercise tolerance (treadmill walking time to onset of ST-segment depression and angina attacks) of about 10% on average, compared with about 22% under active treatment (nisoldipine). In diabetes therapy, placebo produced no improvement in fasting and postprandial blood glucose levels compared with active treatment (acarbose), and also had no effect on HbA1C values. ADVERSE EFFECTS OF PLACEBO: Adverse drug reactions were observed under treatment with placebo. The frequency and type of placebo-induced adverse reactions also varied between indication groups. For example, typical cardiovascular effects such as tachycardia were observed in the control group. The placebo side effect profile was largely similar to the side effect profile of the active treatment. Some adverse drug reactions (such as "dry mouth" in patients with generalized anxiety syndromes) were observed more frequently under placebo than under active treatment. CONCLUSIONS: Treatment with placebo is frequently effective and cannot therefore be considered as "non treatment". Placebo effects can only be quantified by direct comparison with "non treatment". Like active treatment, treatment with placebo is frequently accompanied by adverse drug reactions. Placebo adverse effects are often disease- and active treatment-specific. The effects and adverse effects of a placebo need to be known before the effects of active treatment in controlled clinical trials can be assessed. The mechanisms of placebo effects are many and varied (e.g. endorphin release, conditioning). Since the use of drugs without regard to evidence-based medicine (prescription of drugs without proven efficacy = pseudoplacebos) may clearly also result in serious adverse effects, such practice may not only be non-beneficial but may even be harmful. PMID- 10367098 TI - [Sudden death in heart failure]. PMID- 10367100 TI - Effects of amlodipine on native cardiac Na+ channels and cloned alpha-subunits of cardiac Na+ channels. AB - The inhibitory effects of amlodipine besilate (CAS 11470-99-6) on the native Na+ current (INa) and cloned human cardiac Na+ channel alpha subunit (hH1) were studied by whole cell patch clamp techniques. Amlodipine produced tonic block of INa in a concentration- and holding potential (HP)-dependent manner with hyperpolarization of H infinity. Amlodipine produced phasic blockade of INa, which was dependent on HP and pulse duration. Amlodipine produced tonic blockade of hH1 in a concentration-dependent manner with 1 : 1 stoichiometry, and phasic blockade of hH1 which was dependent on the pulse duration. Amlodipine blocked INa in a voltage- and frequency-dependent manner via affinity to the resting as well as inactivated conformations of the alpha subunit. PMID- 10367101 TI - Investigation on SCH00013, a novel cardiotonic agent with Ca++ sensitizing action. 1st communication: phosphodiesterase III inhibitory effect and class III antiarrhythmic effect in guinea-pig heart. AB - In order to clarify the mechanism of action of 4,5-dihydro-6-[1-[2-hydroxy-2-(4 cyanophenyl)ethyl]-1,2,5,6- tetrahydropyrido-4-yl]pyridazin-3(2H)-one (SCH00013), a novel cardiotonic agent with Ca++ sensitizing action, its effects on contractile force, atrial rate and action potential, and on the activity of Na+, K(+)-ATPase and phosphodiesterase (PDE) I-IV were studied in the guinea-pig heart. SCH00013 exerted a positive inotropic effect (PIE) on isolated right ventricular papillary muscles in a concentration-dependent manner (EC50 = 9.2 mumol/l): the relative potency was milrinone > SCH00013 > vesnarinone. The PIE of SCH00013 was not influenced by propranolol, a beta-blocker, and SCH00013 did not affect the activity of cardiac Na+, K(+)-ATPase. The PIE of SCH00013 was partially inhibited by carbachol, a muscarinic receptor agonist, which implies a partial contribution of the cAMP-dependent mechanism to the PIE. SCH00013 inhibited the activity of PDE III selectively, but the potency was weak: the IC50 value was 64.9 mumol/l, which was 46 and 3.9 times less potent than those of milrinone and vesnarinone, respectively. SCH00013 and vesnarinone elicited a moderate decrease in the rate of beating of isolated right atria, while milrinone increased it. SCH00013 markedly prolonged the action potential duration and the effective refractory period with no change in the resting membrane potential and dV/dtmax, an indication that SCH00013 may suppress the activity of delayed rectifying K+ channels. These results indicate that SCH00013, that primarily acts as a Ca++ sensitizer, possesses a weak selective PDE III inhibitory effect. The potential positive chronotropic effect of SCH00013 due to PDE III inhibition may be offset by its effect on K+ channels. PMID- 10367102 TI - Investigation on SCH00013, a novel cardiotonic agent with Ca++ sensitizing action. 2nd communication: in vivo cardiovascular effects and bioavailability in dogs. AB - In vivo cardiovascular effects and bioavailability of 4,5-dihydro-6-[1-[2-hydroxy 2-(4-cyanophenyl)ethyl]-1,2,5,6- tetrahydropyrido-4-yl]pyridazin-3(2H)-one (SCH00013), a novel cardiotonic agent, were investigated. In anesthetized dogs, intravenous administration of SCH00013 (0.3-10 mg/kg) increased maximum rate of rise in left ventricular pressure (LVdP/dtmax) in a dose-dependent manner with no change in heart rate (HR) and, at the dose of 3 mg/kg or higher, at which the increase in LVdP/dtmax reached the maximum, it decreased blood pressure. In conscious dogs, oral administration of SCH00013 (1-10 mg/kg) also increased LVdP/dtmax dose-dependently with no change in HR. The increase in the plasma concentration of orally administered SCH00013 (3 mg/kg) was parallel to the increase in LVdP/dtmax. The areas under the plasma concentration versus time curve (AUC0-24 h) after oral and intravenous administration of SCH00013 (3 mg/kg) were essentially identical (15.3 +/- 2.0 micrograms.h/ml and 16.5 +/- 2.1 micrograms.h/ml, respectively). These results suggest that oral bioavailability of SCH00013 is notably high. In conclusion, the positive inotropic effect of SCH00013 with neither elevation of HR nor excessive hypotension, as well as the high oral bioavailability of this compound, may provide a beneficial pharmacological treatment of the patients with congestive heart failure. PMID- 10367103 TI - Investigation on SCH00013, a novel cardiotonic agent with Ca++ sensitizing action. 3rd communication: stereoselectivity of the enantiomers in cardiovascular effects. AB - The cardiovascular effects of the enantiomers, (+)-SCH00013 and (-)-SCH00013, of a novel cardiotonic agent 4,5-dihydro-6-[1-[2-hydroxy-2-(4-cyanophenyl)ethyl]- 1,2,5,6-tetrahydropyrido-4-yl]pyridazin-3(2H)-one (SCH00013) were investigated in vitro and in vivo. The enantiomers of SCH00013 elicited an equipotent positive inotropic effect in isolated guinea-pig papillary muscles. Both of the enantiomers had a modest negative chronotropic effect in isolated guinea-pig right atria and the difference in the chronotropic effects of the enantiomers was not significant. In anesthetized dogs, both enantiomers increased LVdP/dtmax without change in heart rate and slightly decreased blood pressure. These hemodynamic effects of the enantiomers were not significantly different from each other. (+)-SCH00013 and (-)-SCH00013 increased the extent of cell shortening in association with only a small increase in the Ca++ transients in indo-1-loaded rabbit cardiomyocytes, and both the increases in cell shortening and Ca++ transients were not significantly different between the enantiomers. Both isomers equally shifted the relationships between the increases in the cell shortening and Ca++ transients to the left and upward as compared with the relationships for the elevation of extracellular Ca++ concentration and isoproterenol, which indicates that the effectiveness of the Ca++ sensitizing effects of the enantiomers are almost equivalent. The enantiomers of SCH00013 showed equipotent inhibitory effect on the phosphodiesterase (PDE) III activity. The maximal extent and the potency of prolonging effect of the two enantiomers on the effective refractory period were also the same. Thus, the efficacy and potency of the effects on the cardiovascular parameters such as myofibrillar Ca++ sensitivity, PDE III activity and the effective refractory period for the both enantiomers of SCH00013 are equivalent, indicating that the cardiovascular effects of SCH00013 may be due to equal contribution of both enantiomers. PMID- 10367104 TI - Investigation on SCH00013, a novel cardiotonic agent with Ca++ sensitizing action. 4th communication: influence on experimentally induced ventricular arrhythmia in dogs. AB - Influence of 4,5-dihydro-6-[1-[2-hydroxy-2-(4-cyanophenyl)ethyl]- 1,2,5,6, tetrahydropyrido-4-yl]pyridazin-3(2H)-one (SCH00013) and vesnarinone (CAS 81840 15-5) on the arrhythmia experimentally induced by three different methods was investigated in dogs. In digitalis-induced arrhythmia, SCH00013 (3 mg/kg i.v.) showed a tendency to improve the arrhythmia with a decrease in the arrhythmic ratio and an increase in the conducted beats (CB), though these changes did not reach a significant level; it decreased significantly the blood pressure (BP) with no change in the total heart rate (THR) and atrial rate (AR). Vesnarinone (3 mg/kg i.v.) did not affect these parameters except for BP that was decreased significantly. In two-stage coronary ligation-induced arrhythmia, SCH00013 (1 and 3 mg/kg i.v.) did not change the arrhythmic ratio, CB, AR and BP, while the THR being slightly decreased; the arrhythmic ratio showed a tendency to decrease with SCH00013 when examined at 24 h after coronary ligation. Vesnarinone (3 mg/kg i.v.) did not affect these parameters at 24 and 48 h after ligation. In epinephrine (adrenaline)-induced arrhythmia, both SCH00013 and vesnarinone showed exacerbation of arrhythmia. SCH00013 at 1 mg/kg i.v. did not elicit ventricular fibrillation (VF) in five dogs examined, but at 3 mg/kg i.v. it elicited VF in two of three dogs. Vesnarinone at 1 mg/kg i.v. induced VF in all of three dogs examined. Incidence of VF induced by optical isomers of SCH00013 was not significantly different from each other: both isomers elicited VF in two of six dogs at 1 mg/kg i.v. and at 3 mg/kg i.v. each of them induced VF in two dogs examined. The present results indicate that SCH00013 is a cardiotonic agent that is equivalent to or less arrhythmogenic than vesnarinone in animal models of arrhythmia, such as adrenaline- and digitalis-induced arrhythmia and the two stage coronary ligation-induced arrhythmia. Optical isomers of SCH00013 were essentially equieffective in eliciting exacerbation of adrenaline-induced arrhythmia in the dog. PMID- 10367105 TI - Studies on the kinetics, metabolism and re-utilisation after intra-articular administration of hyaluronan to rabbits. AB - The pharmacokinetics of hyaluronan (HA) (CAS 9004-61-9) was measured after intra articular (i.a.) injection of 1 mg/kg of 14C-glucose-labelled HA (14C-HA) (i.e. 2,000 kDa) into the knee joint cavity of rabbits. The movement of HA from the cavity into the systemic circulation was assessed by measuring its concentration in blood and its residue in the cavity. I.a. HA moved into, and disappeared from, the bloodstream at a first-order rate. Synovial fluid, synovial membrane and articular cartilage specimens were taken 24 and 72 h post-injection, and the molecular weight (MW) of HA fractions were measured by using gel filtration chromatography (GFC) using radioactive fractions of known molecular sizes (i.e. 2,000 kDa, 300 kDa and 50 kDa). The radioactivity distribution of synovial fluid had a peak at a 2,000 kDa-equivalent fraction at both 24 and 72 h post-injection, while synovial membrane had a peak at a 300 kDa-equivalent fraction 24 h post injection and at a 50 kDa-equivalent fraction 72 h post-injection. The radioactivity distribution pattern of articular cartilage showed peaks corresponding to 2,000-300 kDa 24 h post-injection and to 50 kDa 72 h post injection. The in vivo re-utilisation of i.a. HA was investigated by assessing radioactivity in the acid-soluble, lipid and protein fractions of plasma, blood cells and liver 72 h post-injection. Results show that HA was broken down into C1 units (carbon cycle) before being re-used as an in vivo constituent in the body. PMID- 10367106 TI - Pharmacokinetics of NS-49, a phenethylamine class alpha 1A-adrenoceptor agonist. 1st communication: absorption and excretion in rats after a single administration of 14C-NS-49. AB - The absorption and excretion of NS-49 ((R)-(-)-3'-(2-amino-1-hydroxyethyl)-4' fluoromethanesulfonanilide hydrochloride, CAS 137431-04-0), a phenethylamine class alpha 1A-adrenoceptor agonist, were studied in rats after a single administration of 14C-NS-49. In addition, the protein binding of this drug was investigated in vivo and in vitro. After oral administration of 14C-NS-49 (1 mg/kg) to male rats, the radioactivity concentrations in the blood and plasma reached maximums within 1 h, then decreased biexponentially with respective elimination half-lives of 25.4 and 11.9 h. Most of the plasma radioactivity was due to unchanged NS-49, indicating of the poor metabolism of this drug in rats. The results of the in situ absorption study using the intestinal loop method showed that 14C-NS-49 was well absorbed from the small intestine. Systemic availability was high (86%), as determined by a comparison of the areas under the plasma concentration-time curves of unchanged NS-49 for oral and intravenous administrations. Food affected the absorption of NS-49. There were no significant sex-related differences in the plasma concentration profiles after the intravenous administration of 14C-NS-49 (p > 0.05). NS-49 was primarily eliminated by renal excretion, 76% and 62% of the dose being excreted unchanged in the urine after intravenous and oral administrations, respectively. The absorption rate, determined on the basis of the urinary excretion of radioactivity, was 83%, being almost the same as the systemic availability. First pass metabolism of NS-49, therefore, is considered to be very limited in rats. The excretion of radioactivity in the bile within 48 h after the oral administration of 14C-NS-49 (1 mg/kg) was 5.9% of the dose, and the excretion of radioactivity in the exhaled air after the intravenous administration (0.2 mg/kg) was negligible. The percentage of 14C-NS-49 bound to serum proteins in vitro was less than 15% in all the animal species tested. The percentage of radioactivity bound to rat serum proteins after the oral administration of 14C-NS-49 (1 mg/kg) was 16-21%. PMID- 10367107 TI - Pharmacokinetics, metabolism and excretion of megazol, a new potent trypanocidal drug in animals. AB - The pharmacokinetics of megazol (CAS 19622-55-0) was investigated after intraperitoneal and oral administration of the drug (80 mg/kg) to mice. The plasma levels were significantly higher after oral administration of drug than after intraperitoneal route (33.8 micrograms/ml compared with 19.0 micrograms/ml for Cmax, 158714 micrograms.h/l compared with 96057 micrograms.h/l for AUC). When suramin (CAS 145-63-1) was administered 24 h before oral administration of megazol, megazol absorption was accelerated (2 h compared with 4 h for Tmax) but the amount absorbed was lower (19.9 micrograms/ml compared with 33.8 micrograms/ml for Cmax and 95547 micrograms.h/l vs 158714 micrograms.h/l for AUC). In the infected mice previously treated with suramin, all estimated pharmacokinetic parameters of plasma megazol were significantly modified, in particularly an increase in the apparent volume of distribution (5.6 l/kg compared with 0.9 l/kg) with a prolongation of the elimination half-life (3 h compared with 0.7 h) of megazol. Excretion of the total radioactivity of megazol was also evaluated after oral administration of 3H-megazol to rats. Total radioactivity was eliminated predominantly via the urinary route (80%) vs. 10.5% in the faeces, 9.5% remaining in the body 8 days after dosing. When unlabelled megazol was orally administered to rats with absence or presence of suramin, megazol recovered in urine and faeces 72 h dosing was: 55.7%/2% vs 20.6%/1.6%, respectively. In the urine, unchanged megazol was present as characterized by LC MS/MS as well as 4 unknown metabolites. This study indicates that suramin significantly affects the pharmacokinetics of megazol and its elimination. PMID- 10367108 TI - Acute and subchronic toxicity studies of the new quinolone antibacterial agent irloxacin in rodents. AB - Irloxacin (6-fluorine-7-(pyrrol-1-yl)-1-ethyl-1, 4-dihydro-4-oxo-quinolone-3 carboxylic acid, CAS 91524-15-1), a new quinolone antibacterial agent, was administered as a single dose to rats and mice both by oral and intraperitoneal route in oder to study its acute toxicity. Its oral subchronic toxicity was also assessed by treating rats for 4 and 13 weeks. The results obtained showed that irloxacin was well tolerated after single administration in mice and rats, with LD50 values above 2000 and 5000 mg/kg for intraperitoneal and oral administration, respectively. In the oral subchronic toxicity studies, the histopathological examination performed after the 13-week treatment period confirmed the kidney as the target organ for toxicity. Increased presence of lipofuscin in the kidneys was observed in animals receiving 2000 or 450 mg/kg/d, and degeneration and/or dilatation of proximal renal tubules and chronic interstitial nephritis in males receiving these dosages. No histopathological findings were observed in the kidneys of animals receiving 100 mg/kg/d for 13 weeks. Other relevant findings were, presence of dark or cloudy urine with slightly lower pH in animals receiving dosages of 450 mg/kg/d and above, increased urinary protein concentration in animals receiving 2000 or 450 mg/kg/d, and increased plasma urea concentration in those receiving 2000 mg/kg/d. Moreover, increased plasma phospholipids and total cholesterol concentration, and increased liver and kidney weights were observed among treated animals. As a summary, the results have shown that irloxacin has a low acute toxicity in both mice and rats. For repeat oral administration in rats, 100 mg/kg can be considered as the non-toxic effect level after a treatment period of 13 weeks. PMID- 10367110 TI - Preclinical safety pharmacology studies on the rapid-acting insulin analogue insulin aspart. AB - [B28 Asp]-human insulin (insulin aspart, CAS 116094-23-6, X14) is a rapidly absorbed analogue of human insulin currently undergoing development for the treatment of diabetes mellitus. Cardiovascular studies and a range of standard behavioural, neurological and organ function tests were conducted in various animal species to investigate potential secondary pharmacological effects of insulin aspart. Single intravenous injections (to ensure maximum bioavailability) of a range of doses of insulin aspart were compared with a soluble human insulin preparation. The validity of the test systems was always tested with appropriate positive and/or negative (vehicle) control compounds. Secondary effects due to hypoglycaemia were identified by simultaneous administration of a glucose solution along with the highest dose of insulin aspart and the vehicle as negative control. No safety issues were raised by these experiments; insulin aspart and soluble human insulin exhibited similar pharmacological profiles throughout. PMID- 10367111 TI - Investigations on the role of cytochrome b5 and divalent cations in the maximal nifedipine oxidase activity of human liver. AB - Native human cytochrome P4503A4 was most active in nifedipine oxidation when incorporated into a binary phospholipid vesicular system with human NADPH cytochrome P450 reductase. The turnover numbers were estimated to be 17.6 and 19.6 min-1 in the presence of Mg2+ or Ca2+ ions (5 mmol/l) in the test system, respectively. Inclusion of b5 in the vesicular CYP3A4: reductase system results in a slightly lower nifedipine oxidase activity of 16.9 min-1 in the presence of Mg2+ ions. These results demonstrate that b5 is not an essential component in CYP3A4 catalyzed nifedipine oxidation in human liver. PMID- 10367112 TI - [Extrapineal melatonin: its place and role in the neuroendocrine regulation of homeostasis]. PMID- 10367113 TI - [Monoamine level, M-cholino-, and beta-adrenergic reception in the cranial cervical sympathetic ganglion during ontogenesis in desympathectomized rats]. PMID- 10367114 TI - [Effects of opioid peptides on ischemic myocardial arrhythmias during laser irradiation and disturbed heart sympathetic innervation]. PMID- 10367115 TI - [Effect of mesodiencephalic modulation on the spike electric activity of the rat small intestine during massive hemorrhage]. PMID- 10367116 TI - [Effect of parlodel on sleep-wakefulness cycle parameters in rats with experimental MPTP-induced depressive syndrome]. PMID- 10367117 TI - [Myoelectric activity and functional relations in various parts of the gastroduodenal complex during vagotomy and serotonin administration]. PMID- 10367118 TI - [Effect of melatonin on the antioxidant system and free radical lipid oxidation during traumatic shock]. PMID- 10367119 TI - [Effects of hepatoprotective agents containing phospholipids on cytochrome P-450 dependent antitoxic liver function during experimental toxic hepatitis]. PMID- 10367120 TI - [Conformational changes in myocardial actin during cardiac insufficiency caused by toxic allergic myocarditis]. PMID- 10367121 TI - [Effect of blockers of the sarcolemmal anion transporter systems on the development of myocardial edema during ischemia and recovery of the contractile function during reperfusion]. PMID- 10367122 TI - [Annexin-I-induced aggregation of the secretory human neutrophil granules]. PMID- 10367123 TI - [Response of skeletal muscles to anabolic steroid is specific and does not depend on the motor activity regime]. PMID- 10367124 TI - [Temperature dependence of various cryoglobulins]. PMID- 10367125 TI - [Effects of exogenous glucocorticoids on the bone marrow colony-forming activity during cytostatic exposure]. PMID- 10367127 TI - [Clonal succession in the hemopoietic system: number of the primitive stem hemopoietic cells and clone life span]. PMID- 10367126 TI - [Anti-arrhythmia activity of the GABA derivative TZ-50-2]. PMID- 10367128 TI - [Escherichia coli M17: analysis of the adhesive phenotype as a factor of the macroorganism colonization and/or pathogenicity]. PMID- 10367130 TI - [Membrane-attacking complexes and membrane complement inhibitors on the leukocyte surface during combined exposure with meningococcus lipopolysaccharide and complement]. PMID- 10367129 TI - [Morphofunctional characteristics of the placental macrophages in vitro in pregnancies with various outcomes]. PMID- 10367131 TI - [Genetic analysis of changes in sizes of telomeres and anti-oncogenes p53 and Rb in children with pre-B acute lymphoblastic leukemia]. PMID- 10367132 TI - [Adaptive response in the reparation-imperfect gout cells]. PMID- 10367133 TI - [Clinical and prognostic importance of the expression of epidermal growth factor receptors in non-small-cell lung carcinoma]. PMID- 10367134 TI - [Prophylaxis of regional lymph node metastasis with liposomes]. PMID- 10367135 TI - [Suppressor and antineoplastic activity of the bone marrow and spleen cells in aging AKR mice]. PMID- 10367136 TI - [Hemo- and lymphatic circulation in the peribronchial lymphatic nodes during lung inflammation and laser therapy]. PMID- 10367137 TI - [The role of SOD in pathogenesis amyotrophic lateral sclerosis]. PMID- 10367138 TI - [Skin lipids after melatonin administration in rats]. PMID- 10367139 TI - [Effect of homeopathic doses of antibodies to antigen S-100 on the electric parameters of neuronal membranes]. PMID- 10367140 TI - [Adrenal zona reticularis regeneration during hyperthermia]. PMID- 10367141 TI - [Effect of mercazolyl on the thyroid follicular structure in rats]. PMID- 10367142 TI - [Diagnostic methods and prophylaxis of purulent inflammation in postsurgical wounds]. PMID- 10367143 TI - Transitions to independent living after ABI. AB - One of the most challenging questions facing service providers and policy makers alike is the appropriate level of supervision for adults living in the community following a brain injury. In a 3-year province-wide study of people entering the community following brain injury rehabilitation, four individuals (out of 22 studied) made a transition from fully supervised living to lower levels of formal supervision during their first year in the community. The present study seeks to provide more information about these four individuals, the factors that allowed them to move to lower levels of supervision, and the perceived success of that transition. For each participant, the interviews conducted over the 1 year period in the initial study were reviewed in detail for information about independent living. In addition, each participant was interviewed again for this study, along with his significant other and three of the community programme staff who were most closely involved with his transition. To summarize, factors most salient in the success of transition included: (1) Roles and relationships of family and programme personnel; (2) staying away from drugs and alcohol; (3) availability of structured daily activities, including productive activity or community programme; (4) financial management; and (5) emotion and behaviour self-control. Secondary themes related to successful community living also included the availability of transportation and prior experience with community living since the onset of brain injury. These results offer the experience of four individuals in moving towards independent living. As such, they provide a starting point for further discussions of the process of supporting individuals to pursue the ultimate goal of independent living. PMID- 10367144 TI - A validity study of the WAIT in closed head injury. AB - This investigation is a validity study on an instrument designed to assess the cognitive status of CHI patients in the acute phase. The Wolinsky Amnesia Information Test (WAIT) is a questionnaire, orally administered and then scored in a standardized format. Interrater reliability has been previously demonstrated. In this study, the WAIT was compared to the Galveston Orientation and Amnesia Test (GOAT), CT scans, and the Glasgow Coma Scale (GCS). Seventy-five subjects participated in the study, all hospital trauma patients at an urban regional trauma centre, who had been referred to the neuropsychology consult service for cognitive testing. Patients were screened to rule out preexisting developmental, drug, alcohol, or documented prior LOC history. Patients' ages ranged from 18-59, with a mean of 33.45 years. A large majority of the patients (81.3%) were diagnosed with a mild level of CHI. Concurrent, discriminative, and construct validity of the WAIT were investigated, and found satisfactory. The data were submitted to factor analysis in order to uncover underlying constructs. In addition to the 'orientation' and 'amnesia' factors identified through factor analysis, a third factor was found and labelled: 'personal temporal/continuum memory'. Limitations of the study and possible directions for future research are discussed. PMID- 10367145 TI - The role of imagery in sexual arousal disturbances in the male traumatically brain injured individual. AB - Over 50% of individuals who suffer traumatic brain injury (TBI) demonstrate a decrease in sexual arousal post-injury. This study investigated the basis of this loss and hypothesized that it occurred as a consequence of the effect of the injury on cognition: specifically, diminution of the ability to form and manipulate sexually arousing imagery. The study compared 14 male participants who identified themselves as having alteration in sexual functioning following traumatic brain injury with a further 14 non-brain injured participants, case matched to them for age and education. All TBI participants were assessed after 2 years following injury, and had had a loss of consciousness of 3 days or greater. The results indicated that the two groups differed in terms of their performance on the Bett's QMI Scale, the Gordon Test of Visual Imagery Control, the Vividness of Sexual Imagery Scale of the Imaginal Processes Inventory, the State Trait Anxiety Inventory, and the Beck Depression Inventory. After correction for the level of depression by analysis of covariance, the TBI participants still featured lower levels of performance on the Sexual Imagery sub-scale of the Imaginary Processes Inventory. The results indicate that sexual arousal disturbances may exist above and beyond the disturbances to affect associated with the psychosocial effects of the TBI. PMID- 10367146 TI - Standardized memory tests and the appraisal of everyday memory. AB - Although ecological validity of traditional tests of memory has been questioned, use of these tests remains the standard in clinical practice. More recently, however, standardized measures with more emphasis on ecological relevance have been developed. One hundred and nineteen adults with diagnosed brain injuries completed traditional instruments of memory assessment, the Luria Nebraska Neuropsychological Battery Memory Scale (LNNB-M) and the Wechsler Memory Scale Revised (WMS-R). Subjects also completed the Rivermead Behavioural Memory Test (RBMT), an instrument designed to measure everyday memory. Additionally, clinicians rated subjects' day-to-day memory functioning at the rehabilitation facility. Results suggest that RBMT is most accurate in classifying severity of memory impairment as rated by clinicians. The LNNB-M and WMS-R were relatively accurate at classifying severely impaired and unimpaired subjects, but were much less accurate at classifying subjects in the mild and moderate impairment ranges. Implications for interpretation and use of these instruments in rehabilitation settings are discussed. PMID- 10367147 TI - ICD-10 codes detect only a proportion of all head injury admissions. AB - Despite criticism, the codes covering traumatic brain injury under the International Classification of Diseases have been widely used for sample collection in the epidemiological studies of head injury. Data have been collected from an Accident and Emergency department's (A&E's) case register on all head injury admissions to hospitals from a defined geographic area between 1 April 1996 and 31 March 1997, and compared with the list collected from the heath authority's central database by using the ICD-10 codes. Thirty seven per cent of the names in the A&E register appeared in the ICD-10 list, and 41% of the names in the ICD-10 list appeared in the list collected from the A&E department. The incidence of reported head injury was found to be three times higher among the 0 15 years age group compared with the 16-64 years age group (3.3% compared with 1% of the population). Among the 16-64 years age group, the peak rates of admission to hospital were in the months of February and August. Approximately 14% of all the head injury admissions stayed in hospital for more than 72 hours, another 14% stayed between 48 and 72 hours, and the rest (72%) stayed for less than 48 hours. PMID- 10367148 TI - Self awareness: effects of feedback and review on verbal self reports and remembering following brain injury. AB - Brain injury may produce impairments in self awareness. The magnitude of impairment is often determined by comparing patient self reports with self reports of others (report-report) or with patient performance (report performance). This paper presents data on the pattern of a self-awareness deficit in memory functioning exhibited by a brain injury survivor 5 years post-injury. The effects of practice and feedback on reporting-recall differences was examined using single case methodology. Several prospective and retrospective self reports were obtained, to allow an examination of reporting about past or future recall. Results showed that recall improved and the magnitude of report-recall differences were reduced with practice and feedback. PMID- 10367149 TI - Effects of blood glucose levels on performance in activities of daily living: a case example of a diabetic man with an acquired brain injury. AB - Dysfunctional blood glucose regulation and sequelae of acquired brain injury (ABI) can affect behavioural training in brain injury rehabilitation. The relationship is examined between blood glucose levels and performance in three activities of daily living (ADL) skills (showering, toileting, and dressing) in a 21-year-old male with ABI and Type I diabetes mellitus. Multiple daily glucometer readings were obtained both pre- and post-treatment. Skills training involved graduated prompting and reinforcement to develop independence in ADLs. Assessment and teaching occurred initially in hospital, and then was presented at home. Results show a strong negative relationship between daily fluctuations in blood glucose levels and performance; no relationship was found between daily mean levels and performance. Implications for treatment approaches for diabetic individuals with ABI are discussed. PMID- 10367150 TI - Rapid evaluation of S-100 serum levels. Case report and comparison to previous results. AB - The aim of this case report is to describe the time course of S-100 serum levels of a patient, after severe head injury, whose blood sample could be drawn very soon after injury. The results were compared to a group of patients in which a correlation between S-100 serum levels and outcome after traumatic brain injury could be demonstrated. Blood samples were taken on admission (mean 2.3 hours), 6, 12 and 24 hours after trauma and then every 24 hours up until and including the fifth day. The outcome was estimated on discharge using the Glasgow Outcome Scale. The S-100 serum level of the patient described in the case report with a favourable outcome had initially risen to 10.0 micrograms/l and showed a rapid decline. In the previous group, patients with unfavourable outcome had a S-100 serum level of 7 micrograms/l mean concerning the first probe (after 2.3 hours mean) compared to 1.5 micrograms/l mean (after 2.23 hours mean) in patients with favourable outcome (p < 0.05). In comparison to the literature, there seems to be differences regarding the enzyme liberation in stroke and head injury. Therefore, S-100 serum levels need to be interpreted with regard to collection time and underlying pathology. PMID- 10367151 TI - Complementary and alternative veterinary medicine. PMID- 10367152 TI - Concerns about recommending over-the-counter drugs to treat helminthiasis. PMID- 10367153 TI - Need for research in companion animal vaccinology. PMID- 10367155 TI - Veterinary acupuncture. PMID- 10367156 TI - Rabies revaccination for companion animals: Canadian data. PMID- 10367158 TI - Demodicosis in an American bison. AB - An 18-month-old, male American bison (Bison bison) was presented with 7- to 9-mm size nodules periorbital, perineal, and on the ventral surface of the tail. Demodex spp. were identified from the exudate by microscopic examination. Examination 6 mo later revealed that the infestation had nearly cleared without treatment. PMID- 10367157 TI - Short-term influence of prednisone and phenobarbital on thyroid function in euthyroid dogs. AB - The short-term effects of prednisone and phenobarbital on serum total thyroxine (tT4), free thyroxine (fT4), and thyroid stimulating hormone (TSH) were evaluated in euthyroid dogs. Twenty-six beagles were randomly divided into 3 groups receiving, respectively, a placebo, prednisone (1.2 to 2 mg/kg body weight, per os, every 12 hours for 3 weeks), or phenobarbital (1.8 to 3 mg/kg body weight for 1 week, then 2.7 to 4.5 mg/kg body weight, per os, every 12 hours for 2 weeks). Blood samples taken over a 6-week period were assayed for serum tT4, fT4, and TSH. Phenobarbital therapy in our study did not affect serum tT4, fT4, or TSH concentrations. Prednisone therapy, however, significantly decreased serum tT4 and fT4, but did not affect serum TSH concentrations. PMID- 10367160 TI - Subclinical copper accumulation in llamas. AB - A 9-year-old, intact male llama with mild ataxia and generalized malaise of 1 month's duration was euthanized following clinical evaluation. Excessive liver copper concentrations were found in the llama and also in clinically normal herdmates. This case documents multiple animals with increased hepatic stores from standard diets and mineral supplements. PMID- 10367159 TI - Intravenous regional anesthesia (Bier block) in a dog. AB - Intravenous regional anesthesia was used in an adult dog as part of a balanced approach to general anesthesia for amputation of the 4th digit of its right hind limb. It allowed the concentration of isoflurane to be reduced to 0.5%. PMID- 10367161 TI - Congenital renal dysplasia and psychogenic polydipsia in a Bernese mountain dog. AB - Congenital renal dysplasia was tentatively diagnosed, based on ultrasound and an intravenous urogram, in a 5-month-old female with polyuria and polydipsia. Creatinine clearance measurement revealed that the renal dysplasia was not the cause of the polyuria. A modified water deprivation test eliminated other differential diagnoses and confirmed psychogenic polydipsia. PMID- 10367162 TI - Astrocytoma arising at the optic disc in a dog. PMID- 10367163 TI - Tumors in wild adult raccoons from a suburban area. PMID- 10367164 TI - Leptospirosis in dogs. PMID- 10367165 TI - The impact of Canada's animal health status on trade. PMID- 10367166 TI - Cancer and blood coagulation: molecular aspects. PMID- 10367167 TI - Cancer therapy based on p53. PMID- 10367168 TI - Three-dimensional conformal radiation therapy for early prostate cancer. PMID- 10367169 TI - Who's on first? Sequencing chemotherapy and radiation therapy in conservatively managed node-negative breast cancer. PMID- 10367170 TI - Retinoids as antitumor agents: a new age of biological therapy. PMID- 10367171 TI - Survival advantage for prostate cancer patients treated with high-dose three dimensional conformal radiotherapy. AB - PURPOSE: The value of treating prostate cancer has been questioned, and some insist that a survival benefit is demonstrated to justify treatment. Prospective dose-escalation studies with three-dimensional conformal radiotherapy technique have demonstrated improvement in biochemical freedom from disease and local control. We report the outcomes of high-dose treatment with three-dimensional conformal radiotherapy compared with low-dose treatment for biochemical freedom from disease, freedom from distant metastasis, cause-specific survival, and overall survival. PATIENTS AND METHODS: The study design was retrospective, involving pairs matched on independent prognostic variables in which each patient treated with low-dose radiotherapy was matched with a patient treated with high dose radiotherapy. Outcomes were compared for two groups of patients: Group I: Three-dimensional conformal radiotherapy treatment--296 patients treated with more than 74 Gy matched on stage, grade, and prostate-specific antigen level, to 296 patients treated with less than 74 Gy. Group II: Three-dimensional conformal radiotherapy treatment--357 patients treated with more than 74 Gy matched on stage and grade to 357 patients treated with less than 74 Gy. RESULTS: Univariate analysis showed that dose is a significant predictor of biochemical freedom from disease, freedom from distant metastasis, and cause-specific survival for group I and biochemical freedom from disease, freedom from distant metastasis, cause specific survival, and overall survival for group II. Multivariate analysis showed that dose is a significant independent predictor in group I for biochemical freedom from disease and freedom from distant metastasis and for biochemical freedom from disease, freedom from distant metastasis, cause-specific survival, and overall survival in group II. DISCUSSION: These data provide strong support for the definitive treatment of prostate cancer with high-dose (> 74 Gy) three-dimensional conformal radiotherapy. These doses can be safely delivered with three-dimensional conformal radiotherapy techniques. Various institutions and industry must collaborate to expand the technology allowing the use of high dose three-dimensional conformal radiotherapy in the national practice beyond centers of technological excellence. PMID- 10367172 TI - Sequencing of chemotherapy and radiation in lymph node-negative breast cancer. AB - PURPOSE: To conduct a retrospective analysis of chemotherapy and radiation sequencing in lymph node-negative breast cancer patients treated with breast conserving surgery. PATIENTS AND METHODS: Between February 1982 and January 1996, 124 patients with lymph node-negative breast cancer underwent breast-conserving surgery with axillary dissection followed by chemotherapy and radiation therapy. The outcome of 68 patients who received chemotherapy first was compared with that of 56 patients who received radiation first. The two groups were balanced with respect to patient age, tumor stage, margin status, and estrogen and progesterone receptor status. Sixty-two percent of the patients had T1 primary disease. The median follow-up among surviving patients was 44 months for the chemotherapy first group and 61 months for the radiation-first group. RESULTS: There were no statistically significant differences in local control, disease-free survival, or overall survival between the two groups. Five-year actuarial rates for local control for the chemotherapy-first and the radiation-first groups were 100% and 94%, respectively. Five-year recurrence-free rates for the chemotherapy-first and radiation-first groups were 92% and 77%, respectively. The 5-year overall survival rate was 89% for both groups. DISCUSSION: Giving chemotherapy before radiation in lymph node-negative breast cancer did not compromise local control. Given the concerns about increased distant metastases if radiation is given first, the chemotherapy-radiation sequence is recommended. PMID- 10367173 TI - Preliminary phase II clinical and pharmacokinetic study of 9-cis retinoic acid in advanced cervical cancer. New York Gynecologic Oncology Group. AB - PURPOSE: 9-cis retinoic acid (ALRT 1057; 9cRA) is a promising new retinoid that binds to all known retinoic acid receptors (RAR and RXR), potentially providing it with a broader spectrum of biologic activity than either 13-cis retinoic acid or all-trans retinoic acid. It has been shown to be at least as active as all trans retinoic acid as a differentiation-inducing and antiproliferative agent in both in vivo and in vitro tumor model systems. METHODS: The New York Gynecologic Oncology Group undertook a prospective, multi-institutional phase II clinical and pharmacokinetic trial of 9cRA in patients with advanced or recurrent squamous cell or adenosquamous cell carcinoma of the uterine cervix. Patients received daily oral doses of 140 mg/m2 of 9cRA. 9cRA and its metabolites were determined by reversed-phase HPLC in plasma samples drawn at 0.5 to 8 hours. RESULTS: Sixteen patients with advanced or recurrent carcinoma of the cervix were enrolled. Therapy was well tolerated with no unexpected toxicities. There were no complete or partial responses observed, indicating that a response rate of 20% or greater to this agent could be ruled out with 95% confidence. Pharmacokinetic parameters for 9cRA on day 1 were in agreement with previous studies. The area under the plasma versus time curves for 9cRA declined by 69% between days 1 and 8 with daily 9cRA dosing and remained at this low level in those patients evaluated on day 28. 4-oxo-9-cis retinoic acid (4-oxo-9cRA) was identified as a major plasma metabolite of 9cRA. Plasma levels of 4-oxo-9-cRA were initially 71% of those of 9cRA, but in contrast to 9cRA, there was no decline in plasma levels on days 8 and 28. The ratio of the area under the curve for the 4-oxo metabolite relative to that of the parent compound increased from less than 1 on day 1 to approximately 2.4 on days 8 and 28. Thus, despite early induction of its own metabolism, levels of total retinoid metabolites persisted at pharmacologic levels at day 28. CONCLUSIONS: 9cRA with this dose and schedule was inactive in women with advanced carcinoma of the cervix. Despite a decline in plasma levels of 9cRA over time, levels of the 4-oxo metabolite tended to persist. While the 4 oxo metabolite is less potent than the parent compound, these data nevertheless suggest that the lack of clinical activity in this patient population may not be attributable exclusively to suboptimal pharmacokinetic parameters. PMID- 10367174 TI - Related case report: in vivo suppression of thyrotropin by 9-cis retinoic acid. AB - Treatment of a 48-year-old woman with advanced cervical cancer with the synthetic vitamin A derivative, 9-cis retinoic acid (9cRA), resulted in thyroid-stimulating hormone suppression without clinical evidence of thyrotoxicosis, which resolved spontaneously when the drug was withdrawn. 9cRA, which is a pan-retinoid (RAR and RXR) agonist, has previously been implicated in induction of interactions between the thyroid receptor and the retinoid receptor, RXR, with endocrine target organ specificity. Furthermore, 9cRA has been shown to down-regulate thyroid stimulating hormone messenger RNA in a pituitary-specific fashion in a murine model, a finding consistent with the pituitary-restricted thyrotoxicosis observed in our patient. This is the first reported case of thyroid-stimulating hormone suppression by 9cRA and suggests that patients receiving this agent should be monitored for pituitary and thyroid function abnormalities. PMID- 10367175 TI - Cost-effective treatment of women with advanced ovarian cancer by cytoreductive surgery and chemotherapy directed by an in vitro assay for drug resistance. AB - PURPOSE: Epithelial ovarian cancer is the fourth leading cause of cancer-related death in women. Five-year survival is about 25%, and new approaches to the treatment of this disease are dearly warranted. This study was designed to determine the feasibility of using an in vitro assay for drug resistance to guide treatment after cytoreductive surgery. We present preliminary results of this study after a median follow-up of 24 months. MATERIALS AND METHODS: We treated 66 patients with advanced ovarian cancer by use of a combination of cytoreductive surgery and chemotherapy. Patient inclusion criteria included histologic confirmation of epithelial ovarian cancer, International Federation of Gynecology and Obstectrics (FIGO) stage III, no prior chemotherapy or radiation therapy, no coexisting neoplasm, and optimal residual disease (< 2 cm). Malignant tissue from the involved ovary of each patient was tested in vitro for drug resistance, and chemotherapy was directed individually by assay results. On the basis of the assay we treated 19 patients with platinum/paclitaxel (TP) and 47 with platinum/cyclophosphamide (CP). RESULTS: Three-year survival (Kaplan-Meier estimate) was 69%; the 95% confidence interval was 58% to 80%. There was no difference in 3-year survival between the 19 patients treated with TP (66%) and the 47 patients treated with CP (74%). The cost-effectiveness of each treatment option was determined. It cost $4615 to achieve 3-year survival for patients receiving CP and $17,988 to obtain a similar survival with TP. The cost effectiveness of assay-directed therapy was $9768. DISCUSSION: Because of the high recurrence rate and the poor long-term survival of women with advanced ovarian cancer, improved therapies for this disease are needed. After surgical debulking, we used results of an in vitro assay for drug resistance to individually select chemotherapy for the patients in this study. Although the 3 year survival of 69% obtained in the present study appears good compared with previously published studies of optimally debulked patients, the results must be viewed with caution. Patients were not randomized, and differences in prognostic factors, such as tumor grade, patient age, and performance status, could account in part for the higher survival found in the current study compared with previously published studies. Treatment with either CP or TP resulted in equivalent 3-year survival. The cost to achieve 3-year survival with this protocol, including the cost of the drug resistance assay, was $9768. We believe that consideration of costs avoided by the elimination of ineffective treatments, needless toxicity, and loss of quality of life would likely increase the cost effectiveness of assay-directed therapy compared with conventional therapy. This study demonstrates that it is feasible to use an in vitro assay in routine clinical practice to eliminate ineffective chemotherapeutic agents. PMID- 10367176 TI - One-hour paclitaxel infusions: review of safety and efficacy. AB - PURPOSE: Paclitaxel has emerged as one of the most active anticancer agents in clinical oncology. Hypersensitivity reactions encountered in the clinical development of this drug prompted the implementation of premedication regimens and prolonged infusions, later amended to a 3-hour infusion schedule. Now that paclitaxel is frequently used in outpatient therapy, optimum efficiency in delivery is an issue. A 1-hour drug infusion is more efficient for both the patient and the clinic staff and can help reduce administration costs. This article reviews the current experience with 1-hour paclitaxel infusions. METHODS: Published studies using 1-hour paclitaxel infusions, including weekly studies, trials of combination chemotherapy, and combined-modality studies, were reviewed. Studies were evaluated for both efficacy and toxicity. RESULTS AND CONCLUSIONS: Paclitaxel administered by 1-hour infusion as part of weekly or every-3-week treatment regimens is active in a variety of tumors, including breast, ovarian, and lung cancer and carcinoma of unknown primary site. Leukopenia, the most common serious toxicity, is usually manageable without hematopoietic growth factor support. The frequency of neurotoxicity appears comparable for 1-hour and 3-hour regimens, and there is no increased risk of hypersensitivity reactions. Infusion duration has been suggested to be an important predictor of response in some tumor types. Evaluation of this issue using a 1-hour paclitaxel infusion as reference is reasonable. One-hour infusions of paclitaxel should simplify outpatient administration, reduce administration costs, and reduce clinic time for patients. Practical information regarding administration of paclitaxel by 1 hour infusion is provided. PMID- 10367177 TI - Present and future of 5-HT receptor agonists as antimigraine drugs. AB - Serotonin (5-hydroxytryptamine; 5-HT) is thought to play an important role in the pathogenesis of migraine. The discovery of the 5-HT1B/1D/1F agonist sumatriptan constitutes a substantial advance in the acute treatment of migraine, though it displays a number of nonnegligible shortcomings. Today, a number of second generation drugs derived from tryptamine are under advanced clinical development or are about to be marketed worldwide for the acute treatment of migraine. These tryptamine derivatives display partial agonist properties at 5-HT1B/1D receptors. It is not yet clearly established whether these agents represent a major improvement over sumatriptan in therapeutic effectiveness. Most of them also show affinity for 5-ht1F binding sites and have better oral pharmacokinetics than sumatriptan. The acute antimigraine effects of this second-generation of triptans seem to be obtained in largely the same way as with sumatriptan: by cranial vasoconstriction and inhibition of trigeminovascular activation from both peripheral and central projections. Future directions in migraine therapy should focus on agents that exhibit high intrinsic activity at 5-HT1B/1D receptors, offer a good safety profile, and demonstrate long-lasting action which might also be considered in migraine prophylaxis. PMID- 10367179 TI - Epidemiology of neuroleptic malignant syndrome. AB - The authors reviewed the trends in the incidence of neuroleptic malignant syndrome (NMS) in studies recently reported in various countries. Possible reasons for the differences in reported incidences were considered. The authors identified publications in English (and their cross-references) that estimated the incidence of NMS with a retrospective or prospective design. They compared the incidence in studies from the United States with those from other countries. The initial retrospective studies from the United States reported higher incidence rates of NMS than did similar studies from elsewhere. More recent prospective studies from the United States report a much lower incidence. Neuroleptic malignant syndrome remains a rare complication of psychotropic treatment if the syndrome is defined stringently. The high incidence reported in earlier studies in the United States can be explained by retrospective study design, loose diagnostic criteria, adherence to an amorphous "spectrum concept," and clinical practices in vogue. PMID- 10367178 TI - Neuropharmacologic aspects of apoptosis: significance for neurodegenerative diseases. AB - The cause of neuronal death in Parkinson's, Alzheimer's, and other neurodegenerative diseases is not known, except in some hereditary forms of these disorders in which a mutated gene has been identified. Even in these cases, the molecular mechanisms that underlie the loss of specific populations of neurons have not been determined, although it is highly probable that apoptosis is involved. Some of the biochemical events that occur during apoptosis have been elucidated. We focus in this review on the role played by the proapoptotic caspases, the antiapoptotic proteins of the Bcl-2 family, and the apoptosis associated signal transducers such as ceramide, calcium, and reactive nitrogen or oxygen species. The role of the mitochondria and the possible implication of cell cycle regulators will also be addressed. Of particular interest are the endogenous inhibitory mechanisms and the pharmacologic agents that can be used to block apoptosis signaling cascades, because they offer models for the development of therapeutic strategies designed to prevent the evolution of pathologic neurodegeneration. PMID- 10367180 TI - Neurophysiologic and neuropsychologic profiles of lamotrigine in epilepsy. AB - The anticonvulsant potential of lamotrigine (LTG) has been extensively assessed in open and double-blind clinical trials including patients with different types of epilepsy. In this review, the neurophysiologic and neuropsychologic profile of LTG is discussed. The electroencephalographic (EEG) findings reveal that the drug induces a decrease both in frequency and in probability of propagation of the EEG epileptiform abnormalities (interictal and ictal), whereas the background activity appears unmodified. In contrast with the traditional antiepileptic drugs (AEDs), LTG does not affect evoked responses (brainstem auditory evoked potentials and somatosensory and visual evoked potentials), indicating only a minor toxic effect on the nervous system. The neuropsychologic assessment shows that LTG does not alter the cognitive functions. In conclusion, the neurophysiologic and neuropsychologic data confirm the efficacy and safety of LTG and support its clinical use as monotherapy in epilepsy. PMID- 10367181 TI - Novel targets for therapeutic intervention against ischemic brain injury. AB - Ischemic stroke is a major health care problem worldwide, and the mechanisms that lead to ischemic brain injury are not completely defined. In the past few years, the identification of many molecules that participate in neuronal death and particularly in apoptosis, has shed light on the development of neuroprotective therapy. Glycine site antagonism of N-methyl-D-aspartate (NMDA) receptors may offer an alternative means to a blockade of glutamate neurotoxicity, which lacks most side effects associated with competitive and noncompetitive NMDA receptor antagonists. Inflammatory processes executed by some proinflammatory molecules contribute to secondary brain injury. Neutral protease calpain is capable of degrading critical cytoskeletal and regulatory proteins, mainly causing postischemic neuronal necrosis. Caspases, a family of cysteine proteases, are at the heart of the apoptotic pathway. Severe DNA damage induced by oxidative stress or apoptotic stimuli activates poly(ADP-ribose) polymerase, causing a rapid depletion of nuclear NAD pools, cellular energy, and thiols. Inhibition of these inducible molecules is expected to achieve effective neuroprotection without serious side effects because the molecules seem relatively unimportant in normal neurotransmission. In the future, the efficacy of these novel strategies should be confirmed in larger study populations, individually and in combinations, before considering clinical application. PMID- 10367182 TI - Parkinson's disease: a preliminary study of yohimbine challenge in patients with anxiety. AB - In this pilot study, we performed an oral yohimbine challenge in 6 patients with Parkinson's disease (PD) and anxiety or depression, 2 parkinsonian patients without psychiatric illness, and 2 healthy control subjects to determine whether patients with Parkinson's disease and anxiety respond to this adrenergic agent in the same way patients with idiopathic anxiety disorders respond. Given the atypical nature of depression in Parkinson's disease (characterized by prominent anxiety), we also wanted to see if patients with Parkinson's disease and depression (but no history of anxiety) are susceptible to yohimbine-induced panic. Parkinsonian patients with anxiety developed panic attacks at frequencies comparable to primary psychiatric patients with panic disorder. The one patient with PD and a history of major depression alone developed a panic attack. Regardless of their history of anxiety or depression, parkinsonian patients demonstrated a vulnerability to yohimbine-induced somatic symptoms. PMID- 10367184 TI - Myotonia associated with sarcoidosis: marked exacerbation with pravastatin. AB - A 37-year-old man with sarcoidosis developed severe electrical and clinical myotonia while taking pravastatin for hypercholesterolemia. Myotonia associated with sarcoidosis is rare. Pravastatin is associated with myotonia in animals. This case suggests that sarcoidosis and pravastatin, two entities not frequently associated with myotonia, may interact in a synergistic manner to produce severe clinical myotonia in humans. PMID- 10367183 TI - Blepharospasm and apraxia of eyelid opening in lithium intoxication. AB - We present the case of a 72-year-old woman with a history of a bipolar mood disorder chronically treated with lithium. Upon having the dose increased, she developed an acute confusional state accompanied by blepharospasm (BS) and apraxia of eyelid opening. Gait instability with frequent falls, pyramid tract signs, and postural tremor in both hands were also evident. On withdrawing lithium, symptoms remitted within 2 weeks. This patient illustrates that BS and apraxia of eyelid opening may be triggered by lithium overdose. Our case warrants the inclusion of lithium in the list of drugs liable to induce such movement disorders. PMID- 10367185 TI - Safety assessment and potential health benefits of food components based on selected scientific criteria. ILSI North America Technical Committee on Food Components for Health Promotion. PMID- 10367186 TI - [The state of thymus and bone marrow cells after long-term exposure to pulsed infrared laser irradiation of locally irradiated and injured skeletal muscle of rats]. PMID- 10367187 TI - [The effect of dexamethasone and thyroxine on the ratio of membrane and soluble forms of intestinal disaccharidases in ontogenesis of rats]. PMID- 10367188 TI - [Dependence of brain cell membranes structure on the level of vertebrate organization]. PMID- 10367189 TI - Abortion service in Amsterdam: deciding and coping in a liberal system. AB - OBJECTIVES: The Netherlands have liberal abortion laws and contraceptives are easily available. In the AMC, a service protocol is designed to facilitate decision-making. In this context, the following questions arise: Do our abortion clients know what went wrong with their contraceptive behavior? Did they decide for more effective contraception afterwards? How did they feel about their own decision and about our services, which include obligatory counselling? STUDY DESIGN: A questionnaire was sent to 23 consecutive clients, who were willing to cooperate. The questionnaire consisted of a scale with 26 items and two open ended essay-questions. RESULTS: The response rate was 45% (11 complete questionnaires). Seven women had used contraceptives, six knew what had gone wrong, five had decided to improve their contraception, four women had not used contraceptives, out of which two decided to start, seven women found the abortion decision difficult, seven women (not the same) were content with their decision, and one of them felt bereaved. The four who were not, on the whole, content with their decision felt bereaved. Eight appreciated our services, ten had used the essay-questions to tell their own story. One woman explicitly regretted her decision and would have liked more intensive counselling. CONCLUSIONS: Our system does not yet function as it should. The reasons may be psychological (unexplored ambivalence), sociocultural (difficulties in using effective contraception) and cognitive (some women were very young and not adequately informed). On a methodological level, we conclude that open-ended questions on these subjects generate more accurate and subtle information. PMID- 10367190 TI - Large observational trial of a new low-dose oral contraceptive containing 20 micrograms ethinylestradiol and 100 micrograms levonorgestrel (Miranova) in Germany. AB - The aim of this observational trial was to obtain clinical data from a large cohort of women using the new low-dose contraceptive (Miranova) and therefore to eliminate the restrictions which are normally present in clinical studies. Data were gathered in a clinical practice setting regarding cycle control and tolerance from women who were using Miranova (20 micrograms ethinylestradiol and 100 micrograms levonorgestrel). A total of 13,085 subjects were initially evaluated for this observational trial; data were available for 12,843 subjects during the treatment cycles. A total of 10,736 subjects (84.2%) completed all six treatment cycles. The method failure (Pearl index), calculated for 70,796 cycles, was 0.44. Cycle control was considered good during the trial; the length of cycle, duration of withdrawal bleeding and intensity of withdrawal bleeding did not significantly change during treatment. Intracyclic bleeding occurred mainly in the first cycle (31.4%) and declined significantly thereafter to a value of 14.5% in the third cycle. In the sixth (final) cycle, the intracyclic bleeding rate was 7.0%. There were no clinically relevant changes in mean systolic blood pressure, mean diastolic blood pressure or body weight. Miranova was shown to be an effective, well-tolerated oral contraceptive with good cycle control. PMID- 10367191 TI - GyneFix-LNG: preliminary clinical experience with a copper and levonorgestrel releasing intrauterine system. AB - A novel intrauterine contraceptive drug delivery system derived from the conventional GyneFix intrauterine implant system is described and the preliminary results in 22 women are discussed. The first objective of the development of the GyneFix-levonorgestrel system was to reduce menstrual bleeding, whether or not related to the effect of copper, by combining a shortened version of the standard GyneFix implant, having a copper surface area of 200 mm2, with a system for the sustained intrauterine delivery of levonorgestrel. The results of this initial observational study indicate that the GyneFix-levonorgestrel system, apart from being well tolerated, is safe and effective. The levonorgestrel component appears to have a beneficial effect on the amount of bleeding. A study on menstrual blood loss will be carried out to substantiate this assumption. PMID- 10367192 TI - Reason, myths and fantasies: preliminary data and reflections about the Portuguese experience with LNG-IUS-induced hypomenorrhea. AB - OBJECTIVE: To evaluate the efficacy and tolerance of the levonorgestrel-releasing intrauterine system (LNG-IUS) among Portuguese women, with particular emphasis on the implications of changes in bleeding pattern. STUDY DESIGN: Forty healthy premenopausal women (average age 42.7 +/- 4.5 years) were recruited in each one of the three Portuguese centers (a total of 120) where the LNG-IUS was used as a contraceptive method during a maximum period of 5 years. All women had previously given their informed consent and were informed about the eventual changes in bleeding pattern. Serum ferritin levels and hemogram measurements were performed before and 1 year after the introduction of the IUS. The results were analyzed by the Student t test (raw data and paired test). RESULTS: Preliminary data showed a continuation rate of 70% with an excellent efficacy rate (no pregnancies) 1 year after the introduction of the IUS. The hypomenorrhea rate was 35% while 12.5% of the women had amenorrhea. The average increase in serum ferritin level was 13 ng/ml and of hemoglobin 0.4 g/dl. The comparisons of hemoglobin results were statistically significant (p = 0.05, t raw data and p = 0.028, t paired test). Most complaints were related to cultural difficulties in accepting the changes in menstrual bleeding pattern. CONCLUSION: In order to minimize and control the most common menstrual bleeding side-effects that occur when using this IUS, as well as to improve the continuation rate, clinicians must make a judicious choice of potential users, owing to the psychological and cultural implications that menstrual bleeding changes can have, mainly among women of low socioeconomic levels. Moreover, non-contraceptive therapeutic benefits should be emphasized. PMID- 10367193 TI - Safety and effectiveness of hormonal postcoital contraception: a prospective study. AB - OBJECTIVE: The aim of this study was to evaluate the demographic characteristics of the population attending our hospital requesting postcoital contraception and to determine the effectiveness of the method and its side-effects. METHODS: A total of 503 women asking for postcoital contraception were included in a prospective open trial. After filling in a questionnaire dealing with demographic and contraceptive data, we prescribed an ethinylestradiol-levonorgestrel combination (100 micrograms/500 mg for two doses 12 h apart). RESULTS: Only 487 women were available for analysis of demographic data. A further 77 were excluded because they presented irregular menstrual cycles and 55 cases were lost for follow-up. Mean age was 22.6 +/- 5.25 years and 35.9% of cases came to the center within the first 5 h after unprotected intercourse. Only 18.8% had previously asked for postcoital contraception. Breakage of condom was the most common reason for request (81.9%). Two pregnancies occurred in the remaining 355 women. According to Dixon's method 15.5 pregnancies should be expected being the overall efficacy of 87.14%. There were no serious adverse effects. Nausea and vomiting (16.33%) were the most prevalent and 59% of the users menstruated at the expected time whilst menses were delayed in 6% of the cases. CONCLUSION: The combination of ethinylestradiol and levonorgestrel in low doses is an effective and safe method of postcoital contraception. PMID- 10367194 TI - Vaginal misoprostol for abortion at 10-13 weeks' gestation. AB - OBJECTIVE: The effectiveness and safety of misoprostol have been reported for abortion up to 22 weeks' gestation. The objective of this study was to demonstrate the effectiveness and safety of self-administration of misoprostol every 12 h, without the need of postexpulsion systematic curettage, in late first trimester abortions (10-13 weeks' gestation). METHODS: A group of 180 women with gestations from 64 to 91 days, self-administered 800 micrograms of vaginal misoprostol every 12 h for a maximum of three doses without performing postexpulsion systematic preventive curettage. Outcome measures included successful abortion (complete and incomplete abortion without requiring a surgical procedure), side-effects, mean expulsion time and vaginal bleeding. RESULTS: Successful abortion occurred in 153/180 (85%) subjects (95% confidence interval (CI) 79-90). The decrease of hemoglobin was statistically significant (p = 0.0001) but clinically unimportant: 12.1 mg/dl (SD 1.1) before treatment and 11.7 mg/dl (SD 1.1) afterwards. The mean expulsion time for patients who aborted after the first dose was 8.3 +/- 3.6 h (median 8 h, range 2-12 h). Vaginal bleeding lasted 6 +/- 3 days, spotting 7 +/- 3 days and total bleeding 13 +/- 4 days. The median dose of misoprostol administered was 1780 micrograms (range 1400 3000 micrograms). CONCLUSIONS: The high degree of acceptability, its efficacy and the fact that postabortion systematic curettage was not needed make misoprostol a suitable alternative to the currently available methods for termination of pregnancy at 10-13 weeks' gestation. PMID- 10367195 TI - Appropriately trained nurses are competent at inserting intrauterine devices: an audit of clinical practice. AB - OBJECTIVE: Very few nurses in the UK insert intrauterine devices (IUDs). Within King's Healthcare, nurses with the advanced family planning qualification (ENB AO8), who have also undertaken further training in IUD insertion, have done so for many years. This is a controversial issue and our aim was to evaluate whether women who had IUDs inserted by specialist nurses had significantly more problems compared with those inserted by doctors. STUDY DESIGN: This was a prospective, ongoing, cohort study of all IUD insertions in hospital-based family planning clinics over a 3-month period. A medical, gynecological, obstetric and contraceptive history was taken, paying particular regard to the reason for requesting an IUD. Clinical assessment, bimanual examination findings and any difficulties with insertion were noted at the time. Any postinsertion problems were noted at the follow-up visit. RESULTS: Fifty IUDs were inserted, 28 by doctors and 22 by nurses. Sixteen were for postcoital contraception and five were routine reinsertions alter removal of an existing IUD. Eleven nulliparous women had insertions (six by doctors and five by nurses). The specialist nurses did not have to consult the doctor with any problems or difficulties with insertion. Twenty-nine clients attended for their follow-up appointment at 6 weeks. Three IUDs required early removal due to unacceptable bleeding and pain--all three had been inserted by doctors. Two postcoital IUDs were removed following menstruation. Of the remaining patients, eight were noted to have minor problems (divided equally between doctor and nurse insertions). CONCLUSION: This study provides favorable evidence that adequately trained nurses are proficient and safe at IUD insertions, regardless of the woman's parity. IUDs inserted by nurses are also likely to have additional benefits of cost-effectiveness and increased client and nurse satisfaction. Follow-up is ongoing. PMID- 10367196 TI - Double Dutch: looking at the usage of combined pill plus condom in girls under 25. AB - OBJECTIVE: To establish what proportion of young girls who were prescribed the contraceptive pill at Manchester family planning clinics have first remembered, and second followed, advice also to use the condom for protection from sexually transmitted infection. METHOD: An anonymous self-administered questionnaire was issued to all females up to the age of 25 years, who were already using the pill and who attended one of 20 different clinic locations. Questions included duration of present relationship, frequency of condom use and reasons for use or non-use. RESULTS: The age of respondents ranged from 13 to 25 years, with one third in the group of 17-19-year-olds. Out of 104 responses, condoms were used most of the time by 29 girls, occasionally by 42 and never by 33. Only 15 of the 104 knew the phrase 'double Dutch'. Dual use most of the time primarily for sexually transmitted disease protection was reported by 23 girls. Previous treatment for sexually transmitted disease was reported by 14 girls, of whom half never used condoms and only three now used condoms most of the time. In the group of girls using condoms occasionally, 70% were using condoms primarily to cover missed pills, antibiotics, etc. Extrapolating from the number of condoms used in the past month and the fact that 80% of girls obtained their condoms from the clinic, the extra cost to the family planning budget is over 14,000 Pounds (over 22,500 Ecu) per year. CONCLUSION: Our result that 22% of young pill users regularly used condoms to protect from sexually transmitted diseases indicates that the message is understood, but the term 'double Dutch' is not. PMID- 10367197 TI - Access to family planning clinics--a local experience. AB - Teenage pregnancy is unacceptably high in the UK. The family planning services in the London borough of Harrow dramatically increased access to teenagers during a period of financial constraints and difficult political and ideological changes in the 5 years from 1992/93 to 1996/97. This was carried out through reconfiguration of clinical services, retraining the multidisciplinary staff, development of clinical guidelines and proformas, outreach activities, reducing wastage, and income generation. PMID- 10367198 TI - Exploring the vision of family medicine: research, technology, and practice. PMID- 10367199 TI - When a resident is incapacitated. PMID- 10367200 TI - Students as assets. PMID- 10367201 TI - Reading between the lines. PMID- 10367202 TI - Curriculum renewal and a process of care curriculum for teaching clerkship students. AB - BACKGROUND AND OBJECTIVES: A school-wide curriculum renewal led to a new clerkship curriculum that teaches core family practice competencies by focusing on the process of care in generalist practice. The organizing framework consists of five prototypic visits and their encounter tasks: 1) new problem visit, 2) checkup visit, 3) chronic illness visit, 4) psychosocial problem visit, and 5) behavioral change visit. METHODS: The seminars occur at the beginning of the rotation and use active learning techniques. Evaluation includes student perceptions of the seminars and teachers and student performance on a clinical performance examination (CPX). RESULTS: Students rated the usefulness of the seminars and the seminar leaders' teaching behaviors favorably. The CPX checklist scores showed that students could perform most of the behaviors expected for each prototypic visit. The students listed the appropriate encounter tasks nearly half of the time when describing what tasks they tried to accomplish during the CPX cases. The students listed concrete behaviors just over 50% of the time. CONCLUSIONS: The students learned the material presented in the seminars and applied it during the CPX. Students can do most of the behaviors but do not seem to describe the tasks as abstractly as faculty. These results come from one class cohort in one medical school, so the generalizability is limited until further work, including other learners, confirms these findings. PMID- 10367203 TI - Domestic violence education in family practice residencies. AB - BACKGROUND AND OBJECTIVES: This study evaluated the extent of domestic violence (DV) education in US family practice residency programs and compared the results to those of a prior study of the same topic. METHODS: We mailed a four-page survey to the directors of all US family practice residency programs. The survey asked the extent to which the topic of DV in particular and other areas of violence in general are included in the curriculum. RESULTS: Surveys were returned from 298 (65.9%) programs, of which 69.4% of respondents indicated that the extent to which violence education is a formal part of their curriculum is either somewhat or a great deal, and 79.9% responded similarly about DV education specifically. On average, programs provide 4-5 hours of training each year, mostly through didactic lectures. Compared to a previous study, our findings demonstrate an increase in violence education in these programs. CONCLUSION: Our findings demonstrate that family medicine educators have increased the amount of residency curricular time devoted to training on DV. PMID- 10367204 TI - Balint group observations: the white knight and other heroic physician roles. AB - BACKGROUND: This article reports a typology of five roles that resident family physicians on occasion assume when relating to troubling patients presented in Balint group seminars. The five roles include the white knight (my way or no way), the Pogo look-alike (I feel your pain), the missing link (you made me do it), the surrogate (I can help), and the revolutionary (let me show you). Each role reflects a particular physician's coping behavior in the context of a specific troubling relationship and is driven, in large part, by unrealistic professional expectations. The roles intend to perform a heroic function in rescuing or protecting the patient, the family, or the physician from a distressing medical situation. Balint group work provides participants with the opportunity to derive clinically useful meaning from their presentations. Residents begin to imagine a variety of therapeutic (helpful) roles to replace the ones they were induced to fill. This process has implications for practicing physicians and physician teachers for improving patient and doctor satisfaction and well-being. PMID- 10367205 TI - Factors influencing satisfaction for family practice residency faculty. AB - BACKGROUND AND OBJECTIVES: Prior published family medicine faculty satisfaction survey results were performed in 1975, 1984, and 1989. The current survey identified specific factors that contribute to family medicine faculty satisfaction and career decision making. METHODS: We mailed a self-administered questionnaire to a proportionate random sample of family medicine faculty of residency programs identified by a pre-survey of programs. The eight-page survey explored 60 professional, scheduling, compensation, and regional factors as they related to overall satisfaction and career plans. The survey also explored 59 similar factors related to the initial decision to enter academic family medicine. RESULTS: Of 383 respondents (59.2% response rate), 93% felt satisfied overall with their faculty roles. Eighty-six percent felt appreciated in their current program, and 94% reported a positive sense of professional challenge. Satisfaction, appreciation, and challenge were strongly intercorrelated and were also positively related to whether the faculty surveyed planned to stay in their current position. The opportunity to mentor residents, the ability to keep more up to date with medical information, and income level stood out as being the most significant of all the factors in predicting overall satisfaction. CONCLUSIONS: The faculty surveyed indicated high levels of satisfaction, feeling appreciated, and professional challenge. The results of this cross-sectional survey identify factors most related to satisfaction and the initial decision to enter academic family medicine. PMID- 10367206 TI - Clinical factors affecting physicians' management decisions in cases of female partner abuse. AB - BACKGROUND AND OBJECTIVES: This study determined which clinical factors influence Canadian primary care physicians' management decisions in cases of female partner abuse. METHODS: We used a cross-sectional survey design and randomly sampled (n = 2,014) English-speaking Canadian physicians with a primary interest in family or general practice who were practicing in any of the 12 provinces and territories in Canada and who were active in private practice and registered to prescribe. Respondents completed a questionnaire that required them to score management decision plans in response to case scenarios illustrating typical office-based situations that might involve domestic violence. RESULTS: The response rate was 50.7% (n = 1,022). Using forward stepwise regression analysis, the strongest predictor of whether a physician endorsed a management plan in response to violence was whether the woman acknowledged or revealed the abuse. Male physicians were more likely than females to endorse talking with the suspected abuser if he was known to them, regardless of the quality of this patient physician relationship with the abuser. CONCLUSIONS: Decisions about whether to deal with the abuse or the selection of a management plan are not dependent on the severity of the physical abuse and the emotional consequences. Whether a woman acknowledges or reveals the abuse, as well as whether both the male and female patients are in the physician's practice, are predictive of whether a physician's response to a case scenario involves dealing with spousal abuse and how he/she will address it. PMID- 10367207 TI - Medical investigations requested by patients: how do primary care physicians react? AB - BACKGROUND AND OBJECTIVES: We investigated the characteristics of patients who request medical investigations and the type of tests requested to study the manner in which primary care physicians react to these requests. METHODS: The study was conducted within the framework of a national health insurance system. Twelve primary care practices from three randomly chosen clinics with different population characteristics participated in the study. The attending physicians were instructed to ask all patients who presented to the clinics within a 7-month period and requested a medical test to complete a questionnaire, indicating the type of test(s) requested and the reason. The physicians were asked to rate the manner in which the patient made the request, their own reaction to the request, and whether they ordered the tests that were requested. RESULTS: During the survey period, 12,322 patients visited the clinics, of whom 295 (2.4%) were reported by a physician to have requested a medical investigation. More-educated patients were more likely to request tests for disease prevention. The types of tests requested were imaging scans, laboratory (blood) tests, and others. The main reason for the request was symptoms (60%), followed by disease prevention (25%). More than 30% of the requests generated self-reported negative feelings in the physician. Physician compliance with patient requests was not significantly correlated with the reason for the request. Laboratory tests were ordered significantly more often than other types. There was a strong correlation between physicians' compliance with the request and physicians' feelings about the request. CONCLUSION: Our findings raise questions about the frequency with which physicians order tests solely in response to patients' requests and provide information about circumstances in which patients make requests for medical investigations. PMID- 10367208 TI - How to use and interpret interval likelihood ratios. AB - BACKGROUND: Likelihood ratios offer important advantages over sensitivity and specificity for characterizing diagnostic tests. They can capture the magnitude of abnormality of test results, whereas sensitivity and specificity require that the test results be dichotomized into positive or negative. This is an important advantage because many diagnostic tests are measured on continuous or ordinal scales. Posttest probabilities calculated from interval likelihood ratios may be different than those calculated from sensitivity and specificity; clinical decisions derived from the use of likelihood ratios may therefore be different from decisions derived from test results characterized by sensitivity and specificity. This article demonstrates the advantages, use, and interpretation of interval likelihood ratios using the clinical scenario of a young child with a high fever. PMID- 10367209 TI - Research fellowships: a road less traveled. PMID- 10367210 TI - For Mary. PMID- 10367211 TI - [Sexual homicide--a data collection from psychiatric records]. AB - After an analysis of the concerning literature we conceptualized a questionnaire with 124 items, which describes the most common characteristics of sexual murderers. The relevance of these characteristics was examined on the basis of psychiatric records. The hypothesis was, that persons who murdered more than once show these characteristics more evidently than single murderers. We compared 20 psychiatric records about single sexual murderers with those about 10 repetitive sexual murderers. Planned offenses, chronic isolation, narcism and tendency to perversity were found more often in persons that murdered more than once. Especially the psychosocial factors were found less often than in the angloamerican literature. More pronounced were the differences between a group of sadistic murderers compared with nonsadistic murderers, using criteria for sadism extracted from Kafft-Ebings first description (1892), Schorsch and Becker (1977), MacCulloch et al. (1983) and Ressler et al. (1988). A case example illustrates the importance of an elaborated concept of sadism and its relation to personality disorder. PMID- 10367212 TI - [Outcome in borderline disorders. A literature review]. AB - This paper reviews the current state of research results on borderline disorders in terms of course and outcome, variables predisposing to good or poor outcome, suicide rates and the influence of psychotherapeutical and pharmacotherapeutical strategies. It turned out that course and outcome of borderline disorders depend on the applied diagnostic criteria and on the length of the follow-up period. The outcome of the follow-up studies of borderline schizophrenia and of the borderline syndrome according to Grinker was on the whole worse compared to those of borderline personality disorder defined by DSM-III/III-R or DIB according to Gunderson or Kernberg's criteria. Further, it could be shown that the GAS or HSRS values of the short-term follow-up studies (up to five years) ranged from 46.4 to 59.2 points whereas those of the long-term studies with an average period of 13.6 till 20 years were measured in the lower and in the mid-60 s that reflects only mild difficulties in psychosocial functioning. However, the high rate of completed suicide in BPD was to be respected: The most extensive follow-up investigation with the highest trace-rate (PI-500) revealed a suicide rate of 9% till now, and the most lethal combination of circumstances was BPD x MAD x alcohol abuse (suicide rate of 38%). Prognostic factors predisposing to poor outcome were substance abuse, admixture with antisocial and schizotypal elements, chronic hostility and affective instability with depressive and anxious features. Prognostic factors predisposing to good outcome were high IQ, extraordinary talent, high attractiveness, likeability and regular appointments with the Alcoholics Anonymous. Finally, the influence of psycho- and pharmacotherapeutical interventions were controversially debated. Several psychodynamic therapy studies resulted in satisfactory outcome scores concerning a subgroup of patients with personality traits like warmth, likeability, reliability, talent. Behavioral treatment strategies such as dialectical behavior therapy by Linehan significantly diminished parasuicidality and impulsiveness. Psychopharmacotherapy should target predominating psychopathological features: Low-dose antipsychotics against micropsychosis and prolonged severe dissociative symptoms, SSRIs and MAOIs against affective instability, and, lithium, carbamazepine or valproate against severe impulsiveness and aggressiveness. PMID- 10367213 TI - [Frequency of immunologic disorders in acute schizophrenia]. AB - Immunological examinations in schizophrenic patients have shown that there are many alterations in both arms of the immune system, i.e. cellular and humoral activities. The results are quite heterogeneous, as not even all schizophrenics show these pathological changes. Immunological findings are assumed to be etiopathogenetically related to the disease process or to be an epiphenomenon. The present study supposes that immunological alterations as they can be found during the course of schizophrenia may be an indicator for somatic vulnerability or an epiphenomenon. 60 male inpatients, fulfilling DSM-IV criteria of schizophrenia where examined during their acute phases of psychosis and during their phases of clinical improvement, by means of a serological profile including cellular and humoral parameters. The control group consisted of 42 healthy male volunteers. It was the aim of this study to find out if there were (a) overall differences in the immune profiles between patients and control group and (b) differences between different categories of schizophrenic disorder. During the acute phase nearly half of the schizophrenic patients showed pathologic immunological parameters, whereas none of the controls did. During the phase of clinical improvement the number of patients with normal immunological findings predominated. Furthermore there was a difference between the Paranoid and the Disorganized Subtype, the latter showing more immunological abnormalities. The results of this study give further support to the hypothesis that immunological aberrations should not be seen as closely etiopathogenetically related to schizophrenic disorders, but rather as an epiphenomenon (e.g. as a stress marker) and/or as indicators for somatic vulnerability. PMID- 10367214 TI - [Hemiballistic syndrome in a drummer]. AB - We report an acute right-sided functional hemiballistic syndrome of the upper and lower limb combined with functional choreatic symptoms of the right forearm and hand of an 59 year old drummer. After a wide range of diagnostic procedures, no somatic reasons could be delineated. Finally a lifetime associated syndrome could be elucidated. We therefore reviewed possible diagnostic and methodical neurological and psychosomatic aspects of functional hemiballism. PMID- 10367215 TI - [Methodology of psychiatric case histories]. AB - This paper deals with the methodology of psychiatric case histories. Three types of case histories are differentiated. The didactic case history teaches about the typical aspects of a psychiatric disorder or treatment by using an individual patient as an example. In the heuristic case history the individual case gives rise to challenging established concepts or to generate new hypotheses. Such hypotheses drawn from inductive reasoning have then to be tested using representative samples. The focus of hermeneutic case histories is the significance of pathological behaviour and experience in the context of the biography of an individual patient. So-called psychopathographies of important historical figures can also be differentiated according to these types. Based on these methodological considerations, quality standards for the named types of case histories are stated. PMID- 10367216 TI - Controversies in coronary stenting. Debate: conditional stenting is the way to go. 20th Meeting of the European Society of Cardiology (Vienna, 22 August 1998) PMID- 10367217 TI - [Genetic counseling in cardiomyopathies]. PMID- 10367218 TI - Nonsustained ventricular tachycardia as a predictor for sudden death in patients with idiopathic dilated cardiomyopathy. The role of amiodarone treatment. AB - BACKGROUND: Sudden death frequently occurs in patients with idiopathic dilated cardiomyopathy. Ventricular arrhythmias are encountered in almost all cases. The prognostic significance of life-threatening arrhythmias such as successfully resuscitated ventricular fibrillation and sustained ventricular tachycardia is well known, while it is controversial for ventricular arrhythmias of a lower degree. Amiodarone has been used widely in these patients but its value in preventing sudden death is still uncertain. The aim of this study was to evaluate the prognostic significance of runs of nonsustained ventricular tachycardia (NSVT) as a hallmark for sudden death and the efficacy of amiodarone in preventing sudden death and reducing overall mortality in a large series of patients with dilated cardiomyopathy. METHODS: Over the period between 1983 and 1994, a series of 151 consecutive patients with idiopathic dilated cardiomyopathy underwent ambulatory electrocardiographic monitoring for a mean period of 191 hours/patient. Seventy-nine patients (56 male, mean age 50.7 +/- 13.1 years) (group A) had ventricular arrhythmias of Lown class < or = 4A, while 72 (53 male, mean age 48.6 +/- 12.8 years) (group B) had one or more NSVT runs. The two groups were well matched in terms of clinical features. Mean follow-up period was 86.8 +/- 38.7 and 74.7 +/- 39.5 months, respectively. In group A no antiarrhythmic drug was administered, while in group B 54/72 patients were treated with amiodarone (mean dosage 300 mg/day) for a mean period of 69.7 +/- 37.8 months (group B1). The remaining 18 patients received class I antiarrhythmic drugs, mexiletine (12) and propaphenone (6) for a mean period of 46.1 +/- 29.4 months, because amiodarone was contraindicated (3) or serious side-effects occurred during amiodarone treatment (15), which was discontinued after a mean period of 3.8 +/- 3.1 months (group B2). RESULTS: The cumulative survival probability in the whole population was 86.6% at two years and 65.6% at five years. The rate of sudden death was 6.0% at two years and 18.3% at five years. No statistically significant difference was observed in terms of all-cause mortality or sudden death in the three groups (A, B1, B2). In group B1, amiodarone determined the disappearance of NSVT at Holter monitoring in 50% of patients (27), with no significant difference in the rate of sudden death between the two subgroups. CONCLUSIONS: In unselected patients with idiopathic dilated cardiomyopathy, cardiovascular mortality does not differ between those with NSVT on chronic amiodarone treatment and those without NSVT who have not undergone antiarrhythmic therapy. There was a trend towards a higher overall and sudden mortality rate in patients with NSVT treated with other antiarrhythmic drugs vs patients with NSVT treated with amiodarone, but due to the small size of the first group no significant difference could be calculated. Assuming NSVT as a potential prognostic marker for sudden death, amiodarone treatment may have exerted a beneficial effect in these patients, but this statement is only a presumption due to the limitations of our study. The disappearance of NSVT during amiodarone treatment is not predictive of a reduced rate in sudden death, so that the potential effect of the drug does not appear to be related to the suppression of NSVT at Holter monitoring. PMID- 10367219 TI - Effects of aspirin or picotamide, an antithromboxane agent, in combination with low-intensity oral anticoagulation in patients with acute myocardial infarction: a controlled randomized pilot trial. AB - BACKGROUND AND OBJECTIVE: Combined treatment with antiplatelet drugs and oral anticoagulants seems more effective than monotherapy in the reduction of thrombotic episodes in patients with ischemic heart diseases. We compared the safety and efficacy of two different antiplatelet drugs, aspirin (asa) and picotamide (pico)--a dual antithromboxane agent--in combination with low intensity oral anticoagulation with warfarin or acenocoumarol in acute myocardial infarction (AMI). PATIENTS AND METHODS: Primary endpoint of the study was to compare the incidence of major events (death, reinfarction, postinfarction angina and heart failure) in AMI patients undergoing thrombolytic therapy. In a controlled randomized parallel group pilot study, 101 patients with AMI were enrolled and treated with asa 160 mg/die plus low-intensity oral anticoagulation (target INR: 1.5-2.5) (n = 51) or pico 300 mg/tid plus low-intensity oral anticoagulation (n = 50). Secondary endpoint of the study was to compare the cumulative incidence of major events plus major hemorrhagic episodes defined as macroscopic hematuria, nose bleeding and melena. RESULTS: The two groups were well matched regarding the main demographic and clinical variables. AMI location was anterior in 22 and 20 patients in the pico and asa group respectively, inferoposterior in 27 (pico) and 29 (asa) patients. At the end of the six-month period, major events were observed in 20 patients in the pico group and in 31 patients in the asa group (p < 0.05). The cumulative incidence of major clinical events plus major hemorrhagic episodes was significantly lower in the pico group in comparison with the asa group (28 vs 48; p < 0.001). CONCLUSION: The results of this pilot study suggest that combination therapy with picotamide and low intensity oral anticoagulation could be a safe and effective alternative to aspirin in patient with AMI. This hypothesis should be confirmed by controlled randomized trial with an adequate sample size. PMID- 10367220 TI - [Heart catheterization via the femoral artery with a 4 French and mobilization at 2 hours]. AB - The use of small catheters for cardiac catheterization, as well as for other diagnostic and interventional procedures, can reduce iatrogenic trauma on cardiac and vascular structures. Early patient mobilization may thus reduce both patient discomfort and the length and cost of stays. The performance of 4 French catheters was evaluated in a pilot cohort of consecutive in patients who underwent coronary arteriography with the use of the femoral Judkins technique and who had no restriction to full ambulation. Patients were helped to resume full ambulation two hours after the procedure, and the femoral access site was inspected 24 hours later upon discharge. Coronary arteriography with 4 French catheters was performed in 45 patients (10 women) aged 62 +/- 10 years. In one patient with anomalous origin of the right coronary artery, selective catheterization of the coronary ostium required a catheter style available only in 5 French. In all cases, selective opacification with 4 French catheters was adequate for diagnosis. Forty-three patients were mobilized 115 +/- 10 minutes after the end of manual compression. Hematoma, bleeding or limb perfusion disturbances were absent in all cases upon inspection 22 +/- 4 hours later. This pilot experience indicates that coronary arteriography with femoral 4 French Judkins catheters is technically feasible and that patient ambulation 2 hours later is safe. This data requires confirmation in a larger patient cohort and can lead to new standards for both patient comfort and the use of hospital resources in coronary arteriography. PMID- 10367221 TI - [The management of syncope in the hospital: the OESIL Study (Osservatorio Epidemiologico della Sincope nel Lazio)]. AB - BACKGROUND: While syncope is generally considered a frequent finding in clinical practice, no clear epidemiological evidence is available about the relevance of such an event in the general population of Italy. METHODS: The OESIL Study was designed and undertaken in 15 hospitals of the Italian region of Latium in order to assess the percentage of emergency-room visits and admissions due to syncope, as well as to analyze the in-hospital diagnostic work-up performed for this condition. RESULTS: During a two-month observation period, 781 (372 males and 409 females, mean age 55.2 (22.8 years) consecutive patients came to the emergency rooms of the 15 hospitals included in the investigation due to a syncope spell (0.9% of emergency room visits); 450/781 patients (57.6%) were subsequently hospitalized (1.3% of all admissions): 48.0% of the admissions were admitted to a general medical ward, 29.3% to an observation ward, 13.3% to a cardiology section, 1.6% to a neurology section and 7.8% to other clinical sections (neurosurgery, general surgery). The mean duration of in-hospital stay was 6.9 (5.8 days; range 1-40 days). During the hospitalization period, 93.1% of patients underwent an ECG, 51.0% an EEG, 44.3% a CT scan of the central nervous system, 40.2% an echocardiogram and 19.5% a tilt-test. The syncope spell was considered to have a cardiovascular origin in 33.8% of the cases and a non-cardiovascular in 11.6% of the cases, while the origin was unknown in 54.4% of the cases. CONCLUSIONS: Collected data support the idea that syncope represents a frequent event in the general population and is responsible for a significant percentage of emergency-room visits and hospital admissions. However, the performance of conventional diagnostic work-ups is far from being satisfactory. PMID- 10367222 TI - [Therapy with nitro derivatives and the development of tolerance: a comparative study with stress ECG and dipyridamole ECG]. AB - The ECG stress test represents the most commonly-used technique to evaluate the occurrence of nitroglycerin tolerance. It acts by increasing cardiac O2 demand with resulting insufficient blood flow through a stenotic coronary artery and development of cardiac ischemia. However, other tests are also potentially suitable, such as the ECG-dipyridamole test. The aim of the present study was to evaluate the acute response of ECG-dipyridamole and ECG-stress tests to nitroglycerin. In particular, the development of nitroglycerin tolerance during chronic therapy was evaluated with both tests in patients with stable angina. Eleven patients (8 men and 3 women) with CAD proven by a previous coronarography, a known history of stable angina within at least six months and a positive response to both the tests were studied. At the end of a seven-day wash-out period, all patients were positive to initial ECG-stress and ECG-dipyridamole tests; after 3 days a new evaluation was carried out (Effort 0 and Dip 0) and this confirmed the previous results. We performed a randomized trial in two phases: acute and chronic therapy. In the acute phase, all patients underwent ECG stress and ECG-dipyridamole tests (Effort 1 and Dip 1) in a randomized fashion one day apart, four hours after administration of a 10 mg/24 h nitroglycerin patch. The chronic phase consisted of 25 days of continuous treatment with a nitroglycerin patch. The two tests (Effort 2 and Dip 2) were always repeated after four hours of the morning therapy. Nitroglycerin does not modify the hemodynamic response to dipirydamole in either acute or chronic treatment. Lastly, our data confirm the efficacy of nitroglycerin on stress and dipyridamole tests after acute administration. Nitroglycerin tolerance is confirmed by both tests although with different patterns. ECG stress test showed nitroglycerin tolerance because time to ischemia and max ST deteriorated during chronic therapy. Moreover, the ECG-dipyridamole test showed nitroglycerin tolerance because five patients with a negative acute test (Dip 1) became positive during chronic therapy (Dip 2). PMID- 10367223 TI - [The predictive value of junctional beats during the radiofrequency transcatheter ablation of the slow pathway of the nodal reentry circuit]. AB - BACKGROUND: Junctional beats (JB) are often recorded during slow pathway (SP) radiofrequency (RF) ablation in patients with atrioventricular nodal reentrant tachycardia (AVNRT). Neither the correlation between JBs and SP potentials nor the role of mechanically-evoked JBs has been clarified yet. METHODS: Two hundred eleven consecutive patients, with common AVNRT, underwent RF transcatheter ablation guided by Jackman SP potential searching. If we were unable to record an SP potential or if 4 RF pulses delivered on ideal ablation sites were ineffective, the ablation was carried out on anatomical landmarks. Light pressure was applied with the ablation catheter to each ablation site before RF delivery in order to evaluate the inducibility of JBs. RESULTS: Transcatheter ablation was performed successfully in 209/211 (99%) patients. In 17 (8.1%) patients, no SP potential was recorded. JBs were observed more often delivering RF in the mid septal region, whereas SP potentials were more often recorded at the base of the Koch triangle. The success rate (successful pulses/overall pulses) was higher in the mid-septal (58.6% in M1, 77.8% in M2) than in the postero-septal region (4% in PSC, 16.8% in P1). JBs showed a higher specificity (73.2 vs 5.3%), positive (55.5 vs 24.6%) and negative predictive value (97.3 vs 63.8%) than SP potential in identifying the successful ablation site. Mechanical JBs were evoked in 23 patients on 29 ablation sites, and 18/29 (62.1%) of them were successful ablation sites. CONCLUSIONS: The recording of JBs during or before RF ablation is a useful parameter to guide SP ablation in patients with AVNRT. Although the underlying mechanism has not been clarified yet, their preferential occurrence in the mid septal region suggests that they might be due to thermal stimulation of compact atrioventricular node. PMID- 10367224 TI - Cardiac angiosarcoma: early diagnosis. A case report. AB - Malignant tumours are rare and their diagnostic verification is more frequent at a post-mortem examination. We present a clinical case of a angiosarcoma of the atrium dextrum in a patient where the diagnosis was done precociously. The clinical case we are describing presents some characteristics not noted in literature. The patient did not present any other clinical signs other than giving assumption of the presence of neoplasm and/or secondary localization. The transthoracic echographic exam revealed the methodology to enable the removal of the neoplasm, the outline of which, was better defined with the transesophageal probe. PMID- 10367225 TI - [Chronic bacterial endocarditis on the electrical catheter in a pacemaker wearer: a clinical case report]. AB - This article reports a case of infective endocarditis in a patient with a permanent pacemaker 15 months after the generator had been replaced. The patient had Staphylococcus epidermidis isolated in several blood cultures. No interventional or clinical procedure with any risk of bacteremia was performed, nor was any infective complication of the pocket observed. Thus, the portal of entry of the etiologic agent is unclear. The role of transesophageal echocardiography in detecting pacemaker-induced endocarditis is very important and therapy of choice involves removal of the pacemaker system as soon as possible. PMID- 10367226 TI - [Noninvasive reperfusion tests: myth or reality?]. PMID- 10367227 TI - [Direct stenting of the unprotected common trunk in patients at an elevated surgical risk; the authors' personal experience and a review of the literature]. PMID- 10367230 TI - [2 medicines]. PMID- 10367228 TI - [Glucose-insulin-potassium (GIK) in the reduction of acute myocardial ischemia after an aortocoronary bypass intervention]. PMID- 10367231 TI - Non-atherosclerotic multiple aneurysms of the coronary arteries. PMID- 10367233 TI - Effects of long-term treatment with testosterone enanthate in rhesus monkeys: I. Pharmacokinetics of testosterone, testicular volume and liver metabolism of testosterone. AB - The effects of long-term administration of testosterone enanthate on the pharmacokinetics and bioavailability of testosterone were studied in adult male rhesus monkeys (n = 9), injected with 50 mg of testosterone enanthate (TE) once every 14 days for a total of 32 months. Control animals were injected with 0.2 mL olive oil. Serum testosterone levels increased sharply within 24 h of the first injection of TE and reached a peak on day 3 followed by a sharp decline, but baseline values were not reached even by day 14. Subsequent injections of TE caused a similar pharmacokinetic profile until the 55th injection; testosterone levels on day 3 declined from the 56 to 58th injection and remained in a lower range until the last injection. Repeated injections of TE increased the bioavailability of testosterone as shown by the Area Under the Curve. The nocturnal (22.00 h) surge in testosterone levels during the pretreatment phase was abolished by TE injections. TE injections altered the metabolism of testosterone by the liver, as studied in vitro; while liver from control animals converted testosterone to androstenedione as the major metabolite, androsterone was the major metabolite in chronically TE-treated animals. Spermatogenesis and the associated increase in testicular volume observed in control animals in winter were suppressed in TE-treated animals. The results indicate that repeated TE injections elevate serum testosterone to supra-physiological levels with marked fluctuations in circulating testosterone levels after each injection. Possibly in response to these elevated levels, there was a change in the metabolism of testosterone by the liver as observed in vitro. PMID- 10367232 TI - A re-appraisal of the post-testicular action and toxicity of chlorinated antifertility compounds. AB - Some 30 years ago, alpha-chlorohydrin and some analogues were considered as close to the ideal contraceptive which acted rapidly and reversibly on the post testicular maturation of spermatozoa. Despite their early promise, research funding was withdrawn only 5 years later because of what were considered to be unacceptable side-effects in primates. The literature on the toxic effects of these contraceptive agents was reviewed and was found to be wanting in respect to the rigour of scientific methods applied (impure compounds were used, inappropriate target populations were studied, excessive doses were employed, abstracts were cited from which no full publications subsequently arose). These compounds remain the closest approach yet to non-hormonal contraceptives for males and have led to the synthesis of related compounds which have a similar antifertility action but with much diminished toxicity. If toxicity remains a problem, a range of other compounds now known to have a similar antifertility action, should be investigated. PMID- 10367234 TI - Screening for microdeletions on the long arm of chromosome Y in 53 infertile men. AB - About 30% of couple infertilities are of male origin. They appear in some cases de novo and are considered idiopathic. The aim of our work was to evaluate, in these cases, the prevalence of microdeletions of the long arm of chromosome Y, within the AZF a, b and c regions using molecular biology techniques. Men with azoospermia or oligozoospermia resulting from hereditary, endocrine or obstructive causes, or with a constitutional cytogenetic abnormality were excluded. Fifty-three infertile men with azoospermia or oligozoospermia, as determined by a spermiogram, were studied. Of these, 34 were idiopathic and 7 exhibited a past history of genital infection or biological abnormalities, suggesting partial obstruction of the genito-urinary tract. A further 8 men had a varicocele and 11 cases with a history of cryptorchidism were also studied. Peripheral blood DNA was extracted from each patient, then amplified by multiplex PCR with STS genomic markers from the three Y chromosome AZF zones. PCR products were then analysed on agarose gels. In view of the difficulty of confirming the absence of a signal in molecular biology, each case suspected of having a deletion was checked by multiplex PCR through coamplification with the SRY marker. Five men with microdeletions of the long arm of the Y chromosome were diagnosed among the 53 patients. All of them included the AZFc zone and the intragenic DAZ gene markers. Furthermore, a larger Y chromosome deletion encompassing the 3 AZF zones was diagnosed, and confirmed by cytogenetic analysis. All Y chromosome microdeletions were observed in the 34 truly idiopathic azoospermia/oligozoospermia cases, corresponding to a proportion of 14.7% (or 9.4% considering the whole population of 53 infertile men). The relatively high proportion of microdeletions found in our series suggests the need for strict patient selection to avoid unnecessary screening for long arm Y chromosome microdeletions. PMID- 10367235 TI - Effect of freeze-thawing procedure on chromatin stability, morphological alteration and membrane integrity of human spermatozoa in fertile and subfertile men. AB - Cryopreservation is known to impair sperm motility and decrease the fertilization rate by detrimental effects on acrosomal structure and acrosin activity. However, the consequences of cryopreservation on the integrity of the sperm nucleus, chromatin stability and centrosome are less clear. The present study was designed to determine the effect of the freeze-thawing procedure on chromatin condensation (aniline blue staining) and the morphology (strict criteria) and membrane integrity of human spermatozoa. The structural and functional characteristics of the sperm plasma membrane were measured by the eosin-test and hypo-osmotic swelling test which were done separately. Sperm cryopreservation was performed on semen samples from two groups of men classified as fertile (n = 20) and subfertile (n = 72), based on their reproductive history and semen analysis according to WHO guidelines. The mean percentage of condensed chromatin, morphologically normal spermatozoa and membrane integrity in all semen samples investigated (n = 92) decreased significantly (p = 0.0001) after freeze-thawing, in comparison to the value observed prior to freezing. By comparing the semen samples between fertile and subfertile patients, significantly (p = 0.0009) greater damage was demonstrated in the subfertile than in the fertile group. Furthermore, no significant difference was observed between the two groups with regard to the morphological alteration and structural as well as functional damage of the sperm membrane. In conclusion, the freeze-thawing procedure significantly affects chromatin structure and sperm morphology, especially in the head and the tail regions, and this may explain the lower fertilization rate and IVF/ICSI outcome when frozen-thawed spermatozoa are used. In addition, this study demonstrates that chromatin condensation is a sensitive parameter for the evaluation of cryodamage of semen samples from fertile and subfertile patients, though subfertile patients with very poor semen characteristics have yet to be studied. It is therefore recommended that chromatin condensation be used as an additional parameter for the assessment of sperm quality after freeze-thawing. PMID- 10367236 TI - The mean of sperm parameters in semen donations from the same donor. An important prognostic factor in insemination. AB - We analysed 12,100 consecutive cycles of artificial insemination by donor spermatozoa in 1901 infertile couples. In our analysis, particular attention was given to finding an appropriate way of taking into account the respective effects of female and male factors on the pregnancy success rate and the level at which these factors act (cycle vs. woman and donation vs. donor). A total of 1213 pregnancies occurred. The pregnancy rate per cycle was lower as the age of the woman increased (p < 0.0001) and varied with the type of infertility: fecundity was higher (p = 0.03) in the case of azoospermia than of severe oligozoospermia. After taking into account these factors, significant unexplained variation in likelihood to conceive remained. A part of this heterogeneity was shown to be due to variation in fecundability between semen donors. In order to explain this heterogeneity between donors, compositional covariates were used, particularly the mean of results of the semen analysis performed for donations from the same donor. For each semen characteristic, the overall mean of the different donations of a donor was an important predictive factor of successful insemination: after taking into account all of the other factors, the odds ratios for an increase of 50 x 10(6)/mL spermatozoa, of a 20% increase in sperm motility and of a 2 point increase in the post-thaw quality index, were, respectively, 1.13, 1.37 and 1.56. After adjustment for these factors, the specific characteristics of each semen donation were no longer significantly predictive of successful insemination. This observation has a biological interpretation: sperm with low parameters but produced by a normally fertile man can have a satisfactory success rate. PMID- 10367238 TI - Semen quality changes among 2343 healthy Slovenian men included in an IVF-ET programme from 1983 to 1996. AB - To determine whether semen quality in Slovenians has changed over 14 years (1983 96), we analysed retrospectively the semen of 2343 healthy men with a normal spermiogram, who were partners of women with tubal infertility included in the IVF-ET programme. Age at semen collection, duration of sexual abstinence, semen volume, sperm concentration, total sperm count, percentage of spermatozoa with progressive motility, and normal morphology were determined. Multiple regression analysis was used to assess the changes in sperm characteristics according to the year of semen collection, year of the man's birth and the duration of sexual abstinence. Semen volume, sperm concentration, sperm count and total sperm motility did not change between 1983 and 1996, whereas between 1988 and 1996 rapid progressive sperm motility decreased by 0.95% per year (p < 0.0001). Semen volume, sperm concentration, and sperm count increased with duration of sexual abstinence. After adjustment for the year of semen collection and duration of sexual abstinence, multiple regression analysis showed that sperm concentration decreased by 0.67% per each successive year of birth (p = 0.03). Thus the sperm concentration decreased from 87.6 x 10(6)/mL in men born in the 1940s to 77.3 x 10(6)/mL in those born between 1956 and 1960. After 1960, sperm concentration was found to increase. In 2343 healthy men, no decline in semen quality, except in rapid progressive motility, was observed in the study period. Lower sperm concentration was found among men born between 1950 and 1960. This could be related to worse socio-economic status, stress or negative environmental factors in this time period. PMID- 10367237 TI - Imipramine for successful treatment of retrograde ejaculation caused by retroperitoneal surgery. AB - Retrograde ejaculation is a known complication following retroperitoneal lymph node dissection. Without further therapy, achieving paternity may be impossible. We evaluated the use of the tricyclic antidepressant imipramine in a new ovarian cycle-dependent dose regime for reversal of retrograde ejaculation in 11 patients desiring fertility. Ten patients with retroperitoneal lymph node dissection performed because of testicular cancer, and one with aortic surgery (thromboendarterectomy) were treated in an open, uncontrolled study with imipramine given at a daily oral dose increasing from 25 to 50 mg for 7 days prior to the planned ejaculation or the expected ovulation of the female partner. In all 11 patients, antegrade ejaculation could be induced (sperm counts: 3.9 276.0 x 10(6)/mL). Major side-effects were not observed, but half of the patients complained of minor degrees of dizziness, weakness, nausea or sweating during medication. Under treatment, two patients with normal sperm concentrations induced a spontaneous pregnancy. One ICSI cycle each was performed in 2 patients, with successful fertilization, out no pregnancy. In conclusion, temporary oral intake of imipramine is an effective and safe treatment to re-establish antegrade ejaculation in patients with retrograde ejaculation following retroperitoneal surgery, thus providing possibilities for paternity either through intercourse or by assisted fertilization. PMID- 10367239 TI - GH/IGF-I axis in azoospermia in primary and secondary hypogonadism: a study before and during replacement therapy. AB - The growth hormone/insulin-like growth factor-I (GH/IGF-I) axis was studied in 15 azoospermic patients and in 10 control men. Eight patients were affected by hypergonadotrophic hypogonadism and 7 by hypogonadotrophic hypogonadism. All were studied before and during replacement therapy with testosterone and gonadotrophin, respectively, using the alpha 2 adrenergic agonist, clonidine (clonidine test). The data demonstrate no differences in basal levels for IGF-I and for the GH response to clonidine in azoospermic patients, affected by primary and secondary hypogonadism, before and during replacement therapy when compared with control fertile men. In contrast to some studies which describe a reduced GH response in azoospermia and oligozoospermia, we conclude that basal serum levels of IGF-I and the GH response to clonidine are not impaired in azoospermic patients affected by primary hypogonadism before and after the restoration of normal androgenization, and in azoospermic patients affected by secondary hypogonadism, both before and after restoration of spermatogenesis. PMID- 10367240 TI - Influence of seminal plasma on cryopreservation of human spermatozoa in a biological material-free medium: study of normal and low-quality semen. AB - The objective was to evaluate the efficiency of a biological material-free medium and the role of seminal plasma (SP) in the cryopreservation of human spermatozoa. Normal semen samples and low-quality semen samples were used for this study. After centrifugation of 300 microL fractions of whole semen, pellets were resuspended either in autologous SP or in a chemically defined medium (BM) supplemented or not with 3% bovine serum albumin (BSA); after 15 min at 37 degrees C, the samples were diluted (V/V) with cryoprotective medium (30 mM NaCl; 22 mM sodium citrate, 19.4 mM fructose; 80 mM glutamine; 14%, V/V, glycerol) and maintained for 15 min at room temperature before freezing. Assessment of viability and motility was performed using fresh semen (T0), after centrifugation and resuspension prior to adding the cryoprotectant (T15), after adding the cryoprotectant (T30) and after freezing and thawing (Tpost). In all three resuspending media used, sperm viability and motility (forward and total) decreased (p < 0.05) during both the equilibration period especially before addition of the cryoprotective medium (between T0 and T15) and during the freeze thaw process comparison between T30 and Tpost. The recovery of viable and motile spermatozoa (post-thaw values/values of fresh samples) was higher (p < 0.05) in normal semen than in low-quality semen. In both groups, the recovery was slightly, but significantly, higher with SP than with BM and the presence of BSA has no beneficial effect. To conclude, these data suggest that SP may reduce the deleterious effects of cryopreservation. Nevertheless cryopreservation of spermatozoa in a medium containing neither SP nor biological substances could offer an acceptable cryoprotection of spermatozoa to be used in assisted fertilization procedures, especially for intracytoplasmic sperm injection. PMID- 10367241 TI - The maturation of sperm motility in the epididymis and vas deferens of the vervet monkey, Cercopithecus aethiops. AB - Among the diverse facets of sperm maturation, changes in motility are conspicuous and hence studies of sperm kinematics might provide good indices for sperm maturation. Accordingly, the maturation of sperm motility in the epididymis and vas deferens of the vervet monkey, Cercopithecus aethiops, was assessed using a computer-aided sperm motility analysis system. The results revealed clear trends in the development of both sperm motility per se and in the movement characteristics of motile spermatozoa from different regions of the epididymis, the vas deferens and the ejaculate, reflecting maturational changes associated with the attainment of functional motility and fertility. Motion of spermatozoa from the caput epididymis was sluggish and irregular. As the spermatozoa moved through the corpus epididymis, motility increased sharply, and continued to improve through the cauda epididymis and vas deferens. Despite the high proportion of motile cells, full maturation of motion capabilities was not completed in spermatozoa from the corpus epididymis. Only once spermatozoa had reached the cauda epididymis and vas deferens did they attain their full vigour, and swam rapidly (greater VCL, VSL and VAP) with straightline trajectories (greater LIN, WOB and STR; lower ALH, MAD and CURV). After acquiring their maximal percentage motility and progressive velocity in the cauda epididymis and vas deferens, a slight decline in motility and vigour occurred in ejaculated spermatozoa, and was possibly associated with the ageing of stored spermatozoa. The results from this investigation have revealed clear trends in the maturation of the motility of vervet monkey spermatozoa during their transit through the epididymis and vas deferens and final emergence in the ejaculate, and have provided crucial baseline information on the reproductive physiology of this potentially valuable biomedical model to serve as a reference for future studies in reproductive toxicology. PMID- 10367242 TI - 4th Workshop on Blood Markers in Breast Cancer. Milan, Italy, 28 September 1998. Proceedings. PMID- 10367243 TI - Angiogenesis-related markers and their potential clinical significance. PMID- 10367244 TI - c-erbB-2 in serum of patients with breast cancer. AB - c-erbB-2 is an oncoprotein which is overexpressed in some breast cancers. Recently it has been established that the extracellular domain of c-erbB-2 is shed into the serum of patients with breast cancer. There appears to be no association between tumor stage and extracellular domain of c-erbB-2 (c-erbB 2/ECD): c-erbB-2/ECD seems to correlate with patient prognosis whatever the stage of disease. The data also suggest that c-erbB-2/ECD may be useful in monitoring for tumor recurrence and in predicting resistance to hormonal therapy, but not as useful in predicting response to chemotherapy. This may relate to the power of this marker to reflect disease burden, which has an overwhelmingly negative impact on outcome. PMID- 10367246 TI - Blood-borne tumour markers--a sideways look. PMID- 10367245 TI - c-erbB-2 expression in primary breast cancer. AB - c-erbB-2 is an oncoprotein which is overexpressed in up to 40% of primary breast cancers. c-erbB-2 overexpression is a bad prognostic factor in patients with lymph node-positive disease. Unfortunately, there has been no agreement to date on whether c-erbB-2 overexpression is of prognostic significance in patients with lymph node-negative disease. c-erbB-2 overexpression is correlated with the absence of estrogen receptor expression in a number of publications. Correlation between c-erbB-2 overexpression and hormone sensitivity in the clinical setting is less well established and is the focus of ongoing studies. Both preclinical and clinical studies support an association between c-erbB-2 receptor overexpression and resistance to alkylating agents. In contrast, the data for c erbB-2 and anthracyclines should be viewed in a slightly different manner. Anthracyclines appear to have a greater therapeutic effect in c-erbB-2-positive disease which may be dose sensitive. In c-erbB-2-negative disease not only is the therapeutic effect reduced but there does not appear to be any improved response to higher doses of anthracyclines. The data for c-erbB-2 and the taxanes is still not clear enough to provide any definite conclusions. If there is a correlation it would at present appear to be between paclitaxel and response rates, but this needs to be confirmed in larger studies. Few studies have looked at changes in c erbB-2 on therapy. Those that have seem to show no significant change on either tamoxifen or chemotherapy. PMID- 10367247 TI - Hormonal regulation of MUC1 expression. AB - Several circulating mucinous markers, including CA 15.3, MCA, CA 459, CASA, and Truquant BR, are secreted products of the polymorphic MUC1 gene, and are used as diagnostic tools in patients with breast cancer. In clinical practice the measurement of the levels of these markers in the blood can give important information on the tumor's response to treatment and its biological behavior during disease monitoring. Since the marker levels reflect the activity of the tumor, it is important to know all factors influencing the production/secretion and the blood concentrations of MUC1 mucin. Recent findings suggest that MUC1 gene expression is regulated by steroid hormones and other substances present in the serum. Such observations are very important not only because of their biological significance but also for their clinical implications, as one approach to breast cancer therapy is based on chemical hormone manipulation. Nevertheless, we have preliminarily demonstrated that endocrine treatment in breast cancer patients does not influence the circulating CA 15.3 serum levels, so changes in marker levels are related only to the clinical evolution of the tumor. PMID- 10367248 TI - Review of clinical studies of CA 27.29 in breast cancer management. AB - A critical review of CA 27.29 and CA 15-3 is performed in this paper. A review of the literature is undertaken. A review of the FDA submissions for 27.29 for both early stage and monitoring metastatic breast cancer patients is reviewed. PMID- 10367249 TI - TPA and CA 15.3 measurements for breast cancer monitoring in a routine setting. AB - TPA and CA 15.3 concentrations were routinely determined in serum of patients treated for breast cancer during a 15-month period. ROC curves did not show differences in the ability to differentiate between NED and PD on the basis of matching tumor marker values. During monitoring of patients with NED, TPA levels showed fluctuations of more than 25% that were not disease related. We concluded that CA 15.3 is a more slowly reacting marker of tumor burden than TPA, which is an immediate indicator of cell turnover. PMID- 10367250 TI - TPS monitoring in metastatic breast cancer. AB - The management of metastatic breast cancer patients reflects the heterogeneous nature of the disease. While patients may benefit from hormonal treatment, in most cases more toxic chemotherapy is applied in the advanced stages. The pretreatment levels of TPS in patients with metastatic breast cancer are correlated with prognosis. Decreasing TPS levels (> 50%) during treatment are indicative of response. The fastest decrease in TPS levels is obtained in patients with a favorable prognosis. Increasing TPS levels (> 25%) predict disease progression with a considerable lead time (median 8 months). The clinical impact of these observations is discussed in this paper. PMID- 10367251 TI - Correlation of biochemical tumor markers with conventional pathological features in primary breast cancer. AB - In this prospective study the correlation of pathological with biological prognostic factors and serum tumor markers has been investigated in 574 patients with primary invasive breast cancer. The p53 protein and Bax level correlated positively with tumor size, lymph node status and histological grade. The serum levels of CEA, CA 15.3, TPA-M and TK correlated with tumor extent. There was a significant difference between pre- and postmenopausal breast cancer patients in serum levels of TPA-M and cytosol levels of Bax. Whether these correlations can help in predicting the prognosis of breast cancer by providing additional information with respect to the conventional factors, will have to be investigated by several years of careful clinical follow-up. PMID- 10367252 TI - Chemically defined structured lipids: current status and future directions in gastrointestinal diseases. AB - Over the past two decades various concepts for the supply of lipids in parenteral and enteral nutrition have been developed. Traditionally, the nutritional dietary management typically includes physical mixtures of medium chain and long-chain triglycerides. Recently, chemically defined structured lipids have been developed that combine the advantages of conventional fats with those of special purposes. The first structured lipids were produced by mixing pure medium-chain triglycerides and long-chain triglycerides, allowing hydrolysis to free fatty acids, followed by random transesterification of the fatty acids into mixed triglyceride molecules. This results in a triglyceride containing combinations of short-, medium-, and long-chain fatty acids on a single glycerol backbone. These have unique chemical, physical, and/or physiological properties which differ from simply blending mixtures from the starting fats. By use of 1,3-specific or 2 specific lipases it is now possible to synthesize 1,3-specific or 2-specific triglycerides containing short- and/or medium-chain acids. For instance, incorporation of linoleic, arachidonic, or eicosapentaenoic acid at the sn-2 position is being evaluated for the specific objective of modulating membrane fatty acid composition and essential fatty acid absorption in models of cancer, burns, and immune dysfunction. This contribution reviews the current status of experimental and clinical studies of chemically defined structured lipid-based fat emulsions, with emphasis on their therapeutic potential for nutritional support in hospitalized patients. PMID- 10367253 TI - Is glutamine essential for the maintenance of intestinal function? A study in the isolated perfused rat small intestine. AB - Glutamine has received considerable interest as a gut-targeted nutrient due to its proposed key role in the maintenance of intestinal structure and function. We used a preparation of isolated vascularly perfused rat small intestine to investigate whether glutamine is essential for the maintenance of intestinal function. When glutamine was available, arterial glutamine was extracted at 15 +/ 2%, and net uptake was -89 +/- 5 nmol min-1 g-1. Nitrogenous metabolites ammonia, alanine, and citrulline (41 +/- 7, 41 +/- 4, and 11 +/- 2 nmol min-1 g 1, respectively) were released into the venous perfusate, but only ammonia was also excreted into the lumen (36 +/- 3 nmol min-1 g-1). In the absence of exogenous glutamine alanine release was halved and that of citrulline and ammonia nullified. Additional inhibition of glutamine synthetase yielded the same results. In all cases variables of tissue function were fully maintained also in the absence of exogenous and/or endogenous glutamine. The inhibition of glutaminase/amidotransferase reactions, however, was accompanied by a reduction in glutamine consumption and a graded deterioration in tissue function. In conclusion, glutamine seems to be dispensable as a metabolic fuel to be fully oxidized by the mucosa. However, the inhibition of major glutamine consuming pathways was associated with impaired tissue function and viability. Therefore the role of intestinal glutamine metabolism seems to be threefold: (a) providing affluent amounts of nitrogen precursors for mucosal anabolic pathways to maintain intestinal structure and function, (b) feeding the liver with an optimal substrate mix, and (c) providing citrulline and thereby arginine for the whole organism. PMID- 10367254 TI - Effects of vascular or luminal administration and of simultaneous glucose availability on glutamine utilization by isolated rat small intestine. AB - This study examined whether the route of glutamine administration and the simultaneous availability of glucose affect intestinal glutamine metabolism. We measured net substrate exchange rates of glutamine and its nitrogenous products in the isolated vascularly and luminally perfused rat small intestine (a) as a function of glutamine provision from either the vascular or the luminal or simultaneously from both sides and (b) as a function of simultaneous availability of glucose from various routes. When glutamine was provided from the lumen, only 19-32% of absorbed glutamine appeared intact in the venous effluent, but the release of metabolic products was 170 +/- 5 nmol N min-1 g-1. This measure of intestinal glutamine metabolism was unchanged when glutamine was available only in the vascular perfusate (164 +/- 6 nmol N min-1 g-1). It increased, however, to 271 +/- 14 nmol N min-1 g-1 (P < 0.001) when glutamine was available simultaneously from both the luminal and the vascular perfusate. Glutamine consumption (-110 +/- 6 vs. -70 +/- 5 or -91 +/- 5 vs. -73 +/- 7 nmol min-1 g-1; P < 0.05 each) and the production of citrulline (11.4 +/- 0.7 vs. 10.0 +/- 0.8 or 9.8 +/- 0.5 vs. 7.8 +/- 0.4 nmol min-1 g-1; P < 0.05 each) or ammonia (124 +/- 7 vs. 88 +/- 4; P < 0.01 or 78 +/- 4 vs. 68 +/- 5 nmol min-1 g-1) decreased when glucose (vascular or luminal perfusate) became available in addition to glutamine. We conclude that glutamine is utilized by the small intestine very efficiently regardless of the route of administration being enteral or parenteral. The two routes can be used interchangeably to provide the intestinal mucosa with glutamine. Glucose and glutamine may partially substitute each other, most likely for the purpose as a metabolic fuel. PMID- 10367255 TI - Experience of 1446 rectal cancer patients in Korea and analysis of prognostic factors. AB - In order to investigate the changing pattern of rectal cancers in Korea and to identify prognostic factors, we investigated the case histories of 1446 rectal cancer patients who had received surgical treatment. During the study period there were trends toward a decrease in the ratio of rectal cancer to colon cancer, earlier detection (more Dukes' stages A and B and fewer C), a decrease in the number of abdominoperineal resections, and an increase in the number of sphincter-preserving operations. Univariate analysis of prognostic factors showed that gender, obstruction symptoms, preoperative serum carcinoembryonic antigen (CEA) level, tumor size, depth of bowel wall invasion, lymph node metastases (presence and number), tumor differentiation, operative method, and date of operation were significant, but age, symptom duration, and tumor location were not. The use of sphincter-saving operations did not adversely affect the clinical outcome. Multivariate analysis showed lymph node metastasis factor to be the most significant factor (P < 0.001); the depth of bowel wall invasion, differentiation, CEA level, and date of operation were also significant (0.001 < P < 0.05). This study shows that although anatomical extent of disease (depth of invasion and lymph node metastasis) is the most reliable prognostic predictor in rectal cancer, other factors such as preoperative CEA level and tumor differentiation also provide important information on the outcome and use of an anal-preserving operation does not adversely affect the patient survival. PMID- 10367256 TI - Increased expression of proliferative Ki-67 nuclear antigen is correlated with dysplastic colorectal epithelium in ulcerative colitis. AB - Because patients with ulcerative colitis have an increased long-term risk of colorectal cancer, colonoscopic surveillance with multiple biopsies is commonly performed for histopathological detection of dysplasia to select high-risk patients for prophylactic colectomy. Improved differentiation between neoplastic vs. nonneoplastic changes is needed because active inflammation may cause significant misinterpretation of nonneoplastic reactive/regenerative changes in the epithelium. We investigated whether the expression of proliferative antigens is correlated with various degrees of epithelial dysplasia and inflammatory changes in biopsy specimens from patients with long-standing ulcerative colitis. Colorectal biopsy specimens from patients undergoing colonoscopic surveillance were analyzed immunohistochemically using two types of monoclonal antibodies: MIB 1 against Ki-67 and NCL-PCNA against proliferating cell nuclear antigen for structural, active inflammatory, and dysplastic changes. Specimens from patients without inflammatory bowel disease or neoplasia were used as controls; these showed no increased proliferation. However, increased staining with the MIB-1 monoclonal antibody was detected in 9% of the specimens from patients with long standing ulcerative colitis without active inflammation or dysplasia; this was significantly more common in specimens indefinite for dysplasia, probably positive (24%), and in those with definite dysplasia of low (47%) or high grade (67%; P = 0.008). For increased PCNA staining, there was a non-significant correlation (P = 0.30) with increasing degrees of dysplasia. Increased MIB-1 immunostaining was found in 50% and increased PCNA immunostaining in 75% of the specimens displaying mild inflammation. Both antibodies had a 100% increased staining in specimens with moderate or severe inflammation. Increased proliferation as expressed by MIB-1 is thus better correlated with increasing degree of dysplasia than is PCNA. Neither staining method is able to differentiate neoplastic from inflammatory epithelial changes. However, in the absence of active inflammation, immunostaining for MIB-1 may be a valuable adjunct in the confirmation of dysplastic epithelial changes in long-standing ulcerative colitis, particularly in the indefinite changes category. PMID- 10367257 TI - Manometric analysis of anal sphincter damage after ileal pouch-anal anastomosis. AB - A constant reduction in anal sphincter pressure follows an ileoanal pouch procedure for ulcerative colitis and familiar adenomatous polyposis. We analyzed whether this reduction is more likely due to neurogenic damage or to direct sphincter trauma. Three-dimensional vector volume manometry was performed in 75 patients prior to the ileoanal pouch procedure and 3 months thereafter. Resting pressure was significantly reduced from 83.5 +/- 24.4 to 58.1 +/- 18.0 mmHg and squeezing pressure from 204.7 +/- 63.3 to 173.4 +/- 50.6 mmHg. Moreover, significant vector volume reductions were recorded postoperatively, and the asymmetry index increased significantly (resting: 11.5 +/- 4.1% to 18.4 +/- 7.4%; squeezing: 9.6 +/- 3.1 to 13.0 +/- 6.7%). Functional anal sphincter length at the high-pressure zone remained unchanged. Thus, there was no local damage to proximal or distal anal sphincter segments, which suggests that the postoperative impairment of sphincter function is secondary to neurogenic rather than morphological damage. PMID- 10367258 TI - Influence of phosphotyrosine kinase inhibitors on adhesive properties of highly and poorly metastatic HT-29 colon carcinoma cells to collagen. AB - Organ-specific sites of metastastic lesions are determined in part by integrin mediated adhesion to extracellular matrix (ECM) components. Using poorly (HT-29P) and highly liver-metastatic (HT-29LMM) colon carcinoma cells we previously found different integrin-mediated adhesion to various ECM components, but similar integrin expression of both cell lines. These HT-29 cell lines were used to study adhesion to collagen I (C I) and possible intracellular signaling mechanisms that could explain different adhesive properties. HT-29LMM cells had significantly poorer rates of adhesion to C I (P < 0.05) than HT-29P cells. For examination of the integrin subunits involved in adhesion to C I, cells were treated with various anti-integrin antibodies. These results demonstrated that adhesion of HT 29 cells to C I is mediated in part by the alpha 2 beta 1 integrin. Using immunoprecipitation and Western blotting, both cell lines expressed similar patterns of integrins (alpha 2, alpha 3, alpha 6, and beta 1). Weaker signals were found for the expression of alpha v and beta 5 integrins. Although poorly and highly metastatic cells possessed different patterns of adhesion to C I, these differences were not caused by different expression of integrin subunits. For investigation of the involvement of phosphotyrosine kinases in adhesion, cells were pretreated with the Erbstatin analog, Genistein, or Herbimycin A. Genistein transiently stimulated the adhesive properties of both cell lines. In contrast, Herbimycin A had biphasic effects. At lower concentrations of Herbimycin A stimulation of adhesion was found after 30 and 90 min. However, higher concentrations inhibited adhesive properties. The stimulatory effect was more pronounced in poorly metastatic HT-29P cells. The Erbstatin analog had no effect, probably because of the lack of epidermal growth factor receptor expression in both cell lines. The results suggest that adhesion of tumor cells to ECM components may be dependent on signal transduction into the cell, and tyrosine phosphorylation appears to be involved. PMID- 10367259 TI - High-dose therapy with combined 5-fluorouracil and folinic acid with and without amifostine in the treatment of patients with metastatic colorectal carcinoma. AB - In a prospective clinical trial we treated 27 patients with metastatic colorectal carcinoma a continuous 24-h high-dose regimen of 2.6 g/m2 5-fluoroucil and 500 mg/m2 folinic acid per week for 6 weeks. To evaluate the effect of amifostine, a selective cyto- and radioprotector, in reducing chemotherapy-related mucositis, 740 mg/m2 amifostine was administered before chemotherapy, and the results were compared to 15 patients without amifostine treatment. In the amifostine group the dose was reduced or treatment was stopped because of mucositis observed in 6 of the 12 patients, while 13 of the 15 control patients could not be treated according to the treatment plan. The incidence of gastrointestinal side effects of WHO grade III or higher was reduced in the amifostine group, with no mucositis of this severity, versus 3 of 15 in the control group. The response rate was identical in the two treatment arms (33.3%). PMID- 10367260 TI - Measurement of the anorectal angle by defecography for the diagnosis of fecal incontinence. AB - We assessed the reliability of anorectal angle (ARA) measurement as an index of fecal incontinence. The "posterior" ARA was measured at rest, squeezing, and straining in 69 continent and 82 incontinent subjects all complaining of various evacuation dysfunctions. The two groups were homogeneous with regard to sex distribution (48.6% vs. 51.4% men and 44.7% vs. 55.3% women, n.s.) and age (56.5 +/- 10.2 vs. 59.3 +/- 9.7 years, n.s.). The incidence of rectal prolapse was the same in the two groups (40 each). The intraobserver agreement index from two independent measurements (Pearson's correlation coefficient), age, and gender interaction [T2 Hotelling test in multivariate analysis of variance (ANOVA)] and the most discriminating category of ARA measurement (Fisher's F test in ANOVA) were calculated. In addition, the relationship between ARA and severity of incontinence was assessed by the eta coefficient. Pearson's correlation coefficient was between 0.78 and 0.98 (P < 0.01). The mean ARA differed significantly between the continent and incontinent subjects (104.5 +/- 10.3 degrees vs. 116.2 +/- 23.6 degrees at rest, 84.5 +/- 14.2 degrees vs. 95.1 +/- 20.1 degrees on squeezing, and 133.7 +/- 21.7 degrees vs. 141.7 +/- 25.9 degrees on straining; T2 0.066, P < 0.05 in multivariate ANOVA). No interaction was noted between groups and gender (T2 = 0.023; F = 1.11, n.s.). Resting ARA was shown by ANOVA to be the most discriminating index (F = 9.4 P < 0.01) between the two groups. Overall, ARA measurement was correlated with the severity of fecal incontinence (eta coefficient: 0.894 at rest; 0.811 on squeezing; 0.695 on straining); its accuracy was 79%, the false-positive rate was 15.3% and the false negative rate 26.5%. Irrespective of the underlying abnormality, namely rectal prolapse, ARA measurement by defecography can: (a) be reinterpreted reliably by the same observer and (b) differentiate continent from incontinent subjects. PMID- 10367261 TI - [The CLASSICS Study: ClopidogreL+ASA vs. Ticlopidin+ASA in patients with stents]. PMID- 10367262 TI - [Low-molecular heparin in doses adapted to body weight. Management of thromboembolic diseases]. PMID- 10367263 TI - [Comparative studies on activities of antimicrobial agents against causative organisms isolated from patients with urinary tract infections (1997). III Secular changes in susceptibility]. AB - Susceptibilities to various antimicrobial agents were examined for Enterococcus faecalis, Staphylococcus aureus, Echerichia coli, Klebsiella spp., and Pseudomonas aeruginosa that were isolated from patients with urinary tract infections (UTIs) in 9 hospitals during June 1997 to May 1998, and the results were compared with those obtained during the same period in earlier years. 1. E. faecalis The MIC90s of quinolones for E. faecalis isolated from uncomplicated UTIs have changed better state during the latest period. Among E. faecalis strains, those with high susceptibilities to ampicillin (ABPC) and minocycline (MINO) appeared to had decreased during period of 1995-1997, which recovered during the latest period. 2. S. aureus The sensitive strains of S. aureus to imipenem (IPM) and clindamycin (CLDM) had increased during the period of 1996 1997, but those have decreased again during the latest period. 3. E. coli The susceptibilities of E. coli to MINO have been better in the latest period with the MIC90 was ranged from 2 to 4 micrograms/ml. The susceptibilities to quinolones of E. coli isolated from complicated UTIs had decreased during the period of 1995-1997, but those have recovered during the latest period. 4. Klebsiella spp. Among Klebsiella spp. strains isolated from uncomplicated UTIs, those with low susceptibilities to almost cephems have increased in the latest period. To other antimicrobial agents, the susceptibilities of Klebsiella spp. did not show any changes during the latest period. 5. P. aeruginosa The susceptibilities to most agents of P. aeruginosa did not show any changes, the decreased susceptibilities to cefozopran (CZOP), carbapenems and monobactams of P. aeruginosa observed in 1996 appeared to have been retrieved in 1997. These susceptibility changes should be utilized in determining clinical treatments. PMID- 10367264 TI - [Combination effect of teicoplanin and panipenem on highly resistant strains of MRSA]. AB - We investigated the in vitro combination effect of teicoplanin (TEIC) and panipenem (PAPM) on highly oxacillin-resistant strains of Staphylococcus aureus (MRSA) isolated from various clinical specimens. Combination of TEIC and PAPM using checkerboard titration technique by agar dilution exhibited an excellent effect with mean fractional inhibitory concentration index of 0.18 +/- 0.07 on 47 MRSA strains, and the effects were judged as synergistic against all of the strains tested. In the combination of TEIC and PAPM at 1/4 MIC each against exponentially growing cells of MRSA, good bactericidal activity was found when TEIC and PAPM were added simultaneously, and PAPM was added at 1 or 2 hours prior to addition of TEIC, although the bactericidal activity was scarcely demonstrated when TEIC was added at 1 or 2 hours prior to addition of PAPM. Bactericidal activity against MRSA was enhanced in the combination of TEIC and PAPM at 1/4 MIC each for MRSA than the bactericidal activity of TEIC at 1 MIC alone. TEIC alone showed no bactericidal activity against P. aeruginosa in the mixed cultures with MRSA, while strong bactericidal activity against P. aeruginosa was induced by PAPM. In vitro bactericidal activities against mixed cultures of MRSA with P. aeruginosa were evaluated under conditions of concentrations of TEIC and PAPM, alone and in combination, whose plasma concentrations in human were simulated by a pharmacokinetic simulation model. Bactericidal activity against MRSA was enhanced by the combination of TEIC at 200 mg twice or once daily with PAPM at 500 mg twice daily in comparison with the bactericidal activity of each antibiotic alone, and P. aeruginosa was killed by the antibacterial activity of PAPM. PMID- 10367265 TI - Understanding patient willingness to recommend and return: a strategy for prioritizing improvement opportunities. AB - BACKGROUND: Beginning in April 1995, an ongoing, comprehensive measurement system has been developed and refined at BJC Health System, a regional integrated delivery and financing system serving the St Louis metropolitan area, mid Missouri, and Southern Illinois, to assess patient satisfaction with inpatient treatment, outpatient treatment, outpatient surgery, and emergency care. This system has provided the mechanism for identifying opportunities, setting priorities, and monitoring the impact of improvement initiatives. METHODS: Satisfaction with key components of the care process among 23,361 patients (7,083 inpatients, 8,885 patients undergoing outpatient tests/procedures, 5,356 patients undergoing outpatient surgery, and 2,037 patients receiving emergency care) at 15 BJC Health System facilities was assessed through weekly surveys administered in April 1995 through December 1996. RESULTS: Structural equation models were developed to identify the key predictors of patient advocation-willingness to return for or recommend care. Across all venues of care the compassion provided to patients had the strongest relationship to patient advocation. Within each venue of care, however, a slightly different set of secondary factors emerged. The resulting models provided important information to help prioritize competing improvement opportunities in BJC Health System. In one hospital, a general medicine unit working for several years with little success to improve its patient satisfaction decided to focus on two primary factors predicting patient advocation: nursing care delivery and compassionate care. Root cause analysis was used to determine why two items-staff willingness to help with questions/concerns and clear explanation about tests and procedures-were rated low. On the basis of feedback from phone interviews with discharged patients, the care delivery process was changed to encourage patients to ask questions. Across the next two quarters, this unit experienced significant improvements in both targeted items. DISCUSSION: The significance of compassionate care and care delivery again speaks not only to the importance of the technical quality of clinical care but also to the customer-focused way in which this care was provided. After the primary predictors of patient advocation were identified, management was able to strategically focus improvement initiatives to maximize their impact. Across the organization, improvement teams scanned their data to find key factors where performance was lacking. Once these key opportunities were identified, the teams developed potential solutions and launched initiatives to improve their performance. SUMMARY AND CONCLUSIONS: Results suggest that some core issues are of extreme importance to patients regardless of whether they are receiving care in an inpatient, outpatient, or emergency setting. The compassion with which care is provided appears to be the most important factor in influencing patient intentions to recommend/return, regardless of the setting in which care is provided. PMID- 10367266 TI - Identifying medical center units with disproportionate shares of patient complaints. AB - BACKGROUND: A pilot study was conducted to learn whether an academic medical center's database of patient complaints would reveal particular service units (or clinics) with disproportionate shares of patient complaints, the types of complaints patients have about those units, and the types of personnel about whom the complaints were made. RESULTS: During the seven-year (December 1991-November 1998) study period, Office of Patient Affairs staff recorded 6,419 reports containing 15,631 individual complaints. More than 40% of the reports contained a single complaint. One-third of the reports contained three or more complaints. Complaints were associated with negative perceptions of care and treatment (29%), communication (22%), billing and payment (20%), humaneness of staff (13%), access to staff (9%), and cleanliness or safety of the environment (7%). Complaints were not evenly distributed across the medical center's various units, even when the data were corrected for numbers of patient visits to clinics or bed days in the hospital. The greatest proportion of complaints were associated with physicians. DISCUSSION: Complaint-based report cards may be used in interventions in which peers share the data with unit managers and seek to learn the nature of the problems, if any, that underlie the complaints. Such interventions should influence behavioral and systems changes in some units. SUMMARY AND CONCLUSIONS: Further experience should indicate how different types of complaints lead to different kinds of interventions and improvements in care. Tests of the system are also currently under way in several nonacademic community medical centers. PMID- 10367267 TI - Using quality function deployment to capture the voice of the customer and translate it into the voice of the provider. AB - BACKGROUND: Health care has a number of historical barriers to capturing the voice of the customer and to incorporating customer wants into health care services, whether the customer is a patient, an insurer, or a community. Quality function deployment (QFD) is a set of tools and practices that can help overcome these barriers to form a process for the planning and design or redesign of products and services. The goal of the project was to increase referral volume and to improve a rehabilitation hospital's capacity to provide comprehensive medical and/or legal evaluations for people with complex and catastrophic injuries or illnesses. HIGH-LEVEL VIEW OF QFD AS A PROCESS: The steps in QFD are as follows: capture of the voice of the customer, quality deployment, functions deployment, failure mode deployment, new process deployment, and task deployment. The output of each step becomes the input to a matrix tool or table of the next step of the process. CASE STUDY EXAMPLE: In 3 1/2 months a nine-person project team at Continental Rehabilitation Hospital (San Diego) used QFD tools to capture the voice of the customer, use these data as the basis for a questionnaire on important qualities of service from the customer's perspective, obtain competitive data on how the organization was perceived to be meeting the demanded qualities, identify measurable dimensions and targets of these qualities, and incorporate the functions and tasks into the delivery of service which are necessary to meet the demanded qualities. DISCUSSION: The future of providing health care services will belong to organizations that can adapt to a rapidly changing environment and to demands for new products and services that are produced and delivered in new ways. PMID- 10367268 TI - Lessons from Europe on quality improvement: report on the Velen Castle WHO meeting. AB - BACKGROUND: In conjunction with the German Ministry of Health, the European Regional Office of the World Health Organization (WHO/EURO) held a workshop, "Experiences with Quality Management in an International Context," at Velen Castle, Velen (Nordrhein-Westfalen), Germany, January 15-17, 1998. The approximately 50 participants were selected in part on the basis of recommendations of their respective countries' health ministries. IMPLEMENTATION AND EVALUATION OF QUALITY MANAGEMENT: Possible ways to introduce quality management ranged from introduction of specific process control projects to total quality management (TQM) and reengineering. STRATEGIES FOR IMPLEMENTING QUALITY MANAGEMENT: Working group sessions identified specific strategies for high-level managers, health care providers, and various kinds of consumers to facilitate quality management. For example, managers need to transmit a vision, create a quality management infrastructure, develop reporting structures, establish a system of incentives, and manage the hospital according to the principles of continuous improvement. QUALITY MANAGEMENT MODELS AND TOOLS: Hospitals and other health care providers in Sweden are testing various methods and systems to assess and improve their organizations' ability to meet patients' demands. Benchmarking is being used as a tool for quality management of diabetes care (DiabCare France). The benchmarking data are processed centrally and made available to the health care providers in a user-friendly format for application to their own quality improvement processes. Clinical databases-registries containing process and outcome data for a well-defined patient population-can be used for quality and technology assessment, to answer questions of treatment effectiveness, and as an information tool. PRINCIPLES AND STRATEGIES FOR QUALITY MANAGEMENT AND DEVELOPMENT: Successful implementation of quality improvement benefits from local, professional, and national policies and objectives. A balance of incentives can reward efficiency or specific activities. Laws, rules, and regulations can be useful, especially if used sparingly. More education is needed at all levels of the health care system about how to understand and use information and information systems. Research is needed on what processes result in favorable outcomes. Despite optimism about the cost-saving potential of quality improvement efforts, many interventions are likely to be cost-effective without actually saving costs. Public release of performance data requires careful consideration, with participation of the professions. PMID- 10367269 TI - A. A. Brill and the politics of exclusion. PMID- 10367270 TI - Parochial reading: another reason we talk past each other. PMID- 10367271 TI - Anti-Semitism in the clinical setting: transference and countertransference dimensions. AB - Despite the salient presence of Jews in the history of psychoanalysis, literature on the subject of anti-Semitism in the clinical setting is surprisingly sparse. This paper attempts to comprehend the reasons for the dearth of literature on this important topic. A clinical section then breaks the silence surrounding expressions of anti-Semitism in the consulting room. The major focus is on transference and countertransference reactions that arise with regard to anti Semitism in the clinical setting. Since the first section is concerned with silence in the psychoanalytic community, its focus is primarily on countertransference issues that may hinder the analyst's understanding and use of anti-Semitic material. The second, clinical section focuses on the ways both transference and countertransference reactions combine and influence one another and how they may, when properly attended to, serve as catalytic tools for advancing therapeutic goals. PMID- 10367272 TI - The role of the preconscious in psychoanalysis. AB - The analytic process inevitably involves the interdigitation of the intrapsychic structures of both patient and analyst. This interplay is expressed in transference-countertransference interactions. Drawing a dichotomy between intrapsychic and interpersonal factors as central agents of psychic change is a faulty construction. Affective, behavioral interchanges between patient and analyst reflect their individual intrapsychic organizations and their interplay, which influence the form and nature of psychological change. The safer both patient and analyst feel in relation to each other, the more freely will they relax their customary cognitive controls and permit the emergence of preconscious responses. Preconscious resonance between patient and analyst is likely to facilitate the lifting of repressive barriers and the emergence of unconscious material in both participants. The integration and reworking of old conflicts then becomes possible. The role of the preconscious in facilitating the analytic process is illustrated. Creative use of preconscious processes requires the analyst's self-discipline to preserve the analytic role and keep the treatment safe for both participants. PMID- 10367273 TI - Once more onto the couch: consciousness and preconscious defenses in psychoanalysis. AB - Use of the couch in the analytic situation has a unique impact on the consciousness of both participants in the process. The hypnagogic state of the supine analysand and its resonance with the empathic reverie of the unseen analyst are explored, with a focus not on the contents of analysts' countertransferential associations, but on the diverse but converging modes in which they represent their patients' verbal productions. A clinical example is presented to illustrate the interplay between the patient's and the analyst's imaginations, an understanding of which does away with the false dichotomy between defense analysis and empathic responsiveness. The importance of a patient's conscious and preconscious defenses in the here-and-now transference (suppression, "marginalization," disavowal, negation) is also noted, as is the relation of these defenses to unconscious secondary repression. PMID- 10367274 TI - The distinction between needs and wishes: implications for psychoanalytic theory and technique. AB - After a comprehensive survey of the literature is presented and some caveats entered, this paper delineates the concept of a psychological "need," noting that it bears a complex relation to the concept of a "wish." Need is universal, wish experience-bound. A need, unlike a wish, is not subject to repression. In addition, although a wish can be replaced by another wish, a need cannot be replaced by another need. Whereas the frustration of a wish causes dynamic shifts, the frustration of a need leads to structural disintegration. Needs and wishes can be in harmony or in opposition. The paper also identifies six basic psychological needs, which would seem to be ubiquitous, though the degree to which they are overt and the ways in which they are met vary across cultures. Their gratification seems necessary for healthy psychic development to occur, for relationships to survive, and for psychoanalytic work to take hold and to continue optimally. These needs are (1) the need for one's physical needs to be deemed legitimate; (2) the need for identity, recognition, and affirmation; (3) the need for interpersonal and intrapsychic boundaries; (4) the need for understanding the causes of events; (5) the need for optimal emotional availability of a love object; and (6) the need for a resilient responsiveness by one's love objects under special circumstances. Ordinarily these needs are met during the course of treatment with no deliberate effort by the analyst. In the treatment of some patients, however, they require more direct attention. A number of clinical vignettes are presented to elucidate these ideas. PMID- 10367275 TI - The construction, reconstruction, and deconstruction of memory in the light of social cognition. AB - Psychic trauma results when the ego is overwhelmed by intolerable affect. Some childhood experiences are directly traumatic, requiring no intervening interpretive process to render them traumatic. Troublesome affect that falls short of being truly traumatic, in the strict sense of the term, may nevertheless exert a psychopathogenic effect on the child's psychic development. Whether a child is troubled by an experience depends on what that experience meant to the child. Accordingly, the psychopathogenic effects of childhood experience are a function of the child's general level of cognitive sophistication and specific ability to appreciate the subtle nuances of social interactions. If a child's cognitive capabilities are not up to the task of providing the necessary explanations for a given social interaction, the child is left to fall back on fantasy to fill in the gaps. The field of "social cognition" proves particularly helpful in understanding what a child is capable of gleaning from an experience. Research in this area helps psychoanalysts understand how experience becomes constructed and reconstructed in the form of memories. Social cognition also helps psychoanalysts understand when and how a lived experience ends up being psychopathogenic or, alternatively, ceases any longer to exert an ongoing psychopathogenic effect on an individual's psyche. A review of social cognition research leads to a reconsideration of such psychoanalytic concepts as repression, dissociation, reconstruction, and resistance. It also directs attention to the concept of developmental (as opposed to psychoanalytic) reconstruction and deconstruction. PMID- 10367276 TI - The analytic present in psychoanalytic reconstructions of the historical past. AB - The psychoanalytic reconstruction of specific influential past events has been a regular feature of psychoanalytic practice from its inception. More recently, as reconstructions of incestuous sexual abuse have become more frequent, the reconstructive process has attracted attention, and doubts have been expressed about the validity of the analytic evidence used to substantiate their inference. Over the years, psychoanalytic discourse about the adequacy of this evidence has moved from Freud's uneasy confidence to a rather sharply divided debate between advocates and skeptics. From beyond psychoanalysis, memory research raises questions about the validity of this evidence, and research on influence raises questions about the procedures by which it has been collected. A reexamination of several cases in which reconstructions emerged from unstable therapeutic circumstances leads to the conjecture that dynamics located in the analytic present may have been displaced to the historical past. Although logically "truth in the past" need not foreclose on "truth in the present," in practice, once a reconstruction surfaces, the latter seems to disappear behind the former. PMID- 10367277 TI - Howard Shevrin on Peter Wolff. PMID- 10367278 TI - Hybridization of glass-tethered oligonucleotide probes to target strands preannealed with labeled auxiliary oligonucleotides. AB - In this article we introduce a strategy of preannealing labeled auxiliary oligonucleotides to single-stranded target DNA, prior to hybridization of the DNA target to oligonucleotide arrays (genosensors) formed on glass slides for the purpose of mutation analysis. Human genomic DNA samples from normal individuals and cystic fibrosis (CF) patients (including homozygous delta F508 and heterozygous delta F508/wild type (wt) in the region examined) were used. A PCR fragment of length 138 bp (wt) or 135 bp (mutant) was produced from exon 10 in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, using a new pair of polymerase chain reaction (PCR) primers. This fragment contains four of the most frequent mutation sites causing the disease (Q493X, delta I507, delta F508, and V520F). Each of these mutations was tested using a pair of nonamer (9 mer) probes covalently attached to glass slides, representing the normal (wt) and the mutant alleles. Single-stranded target DNA was isolated from the PCR fragment using one PCR primer labeled with biotin and a streptavidin minicolumn to capture the biotin-labeled strand. Prior to hybridization to the 9-mer array on a glass slide, the unlabeled target strand was preannealed with one, three, or four auxiliary oligonucleotides, at least one being labeled with 32P. As observed previously in several laboratories, the discrimination between normal (wt) and mutant alleles at each site using oligonucleotide array hybridization ranged from very good to poor, depending on the number and location of mismatches between probe and target. Terminal mismatches along the probe were difficult to discriminate, internal mismatches were more easily discriminated, and multiple mismatches were very well discriminated. An exceptionally intense hybridization signal was obtained with a 9-mer probe that hybridized contiguously (in tandem) with one auxiliary oligonucleotide preannealed to the target DNA. The increased stability is apparently caused by strong base stacking interactions between the "capture probe" and the auxiliary oligonucleotide. The presence of the delta F508 mutation was detected with this system, including discrimination between homozygous and heterozygous conditions. Base mismatch discrimination using the arrayed 9-mer probes was improved by increasing the temperature of hybridization from 15 to 25 degrees C. Auxiliary oligonucleotides, preannealed to the single stranded template, may serve several purposes to enable a more robust genosensor based DNA sequence analysis: 1. A convenient means of introducing label into the target DNA molecule. 2. Disruption of interfering short-range secondary structure in the region of analysis. 3. Covering up of redundant binding sites in the target strand (i.e., where a given probe has more than one complement within the target). 4. Tandem hybridization with the capture probe (providing contiguous stacking) as a means for achieving efficient mismatch discrimination at the terminal position of the capture probe (adjacent to the auxiliary oligonucleotide). By use of multiple auxiliary oligonucleotides, all of the above benefits can be derived simultaneously. PMID- 10367279 TI - Mutation detection by stacking hybridization on genosensor arrays. AB - A new strategy for analysis of point mutations using oligonucleotide array (genosensor) hybridization was investigated. In the new approach, a single stranded target strand is preannealed with a labeled "stacking oligonucleotide," and then the partially duplex labeled target molecule is hybridized to an array of glass-tethered oligonucleotide probes, targeted to the region on the target immediately adjacent to the stacking oligomer. In this configuration, the base stacking interactions between the "capture probe" and the contiguously stacking oligomer stabilize the binding of the target molecule to its complementary probe on the genosensor array. The temperature of hybridization can be adjusted so that the target molecule will bind to the glass-tethered probe only in the presence of the stacking oligomer, and a single mismatch at or near the terminal position ol the capture probe disrupts the stacking interactions and thereby eliminates or greatly reduces the hybridization. This stacking hybridization approach was investigated using a collection of synthetic targets, probes, and stacking oligonucleotides, which permitted identification of conditions for optimal base mismatch discrimination. The oligonucleotide probes were tethered to the glass using a simple, improved attachment chemistry in which a 3'-aminopropanol function introduced into the probe during chemical synthesis binds covalently to silanol groups on clean, underivatized glass. "Operating parameters" examined in the stacking hybridization system included length of capture probe, position, type and number of mismatches between the probe and the target, temperature of hybridization and length of washing, and the presence of terminal phosphate group in the probe, at its junction with the stacking oligomer. The results suggest that in the stacking hybridization configuration: 1. Optimal mismatch discrimination with 9-mer probes occurs at 45 degrees C, after which little or no improvement in mispair rejection occurred on lengthy continued washing at 45 degrees C. 2. At 25 degrees C optimal mismatch discrimination occurred with 7- or 8-mer probes, or with 9-mer probes containing an additional internal mismatch. 3. The presence of a phosphate group on the 5'-end of the glass-tethered probe had no general effect on mismatch discrimination, but influenced the relative stability of different mismatches in the sequence context studied. These results provide a motivation for continued development of the stacking hybridization technique for nucleic acid sequence analysis. This approach offers several advantages over the traditional allele-specific oligonucleotide hybridization technique, and is distinct from the contiguous stacking hybridization sitrategy that the Mirzabekov laboratory has introduced (Yershov et al. (1996) Proc. Natl. Acad. Sci. USA 93, 4913-4918; Parinov et al. (1996) Nucleic Acids Res. 24, 2998 3004). PMID- 10367280 TI - Use of a short A/T-rich cassette for enhanced expression of cloned genes in Escherichia coli. AB - A short (43-bp) A/T-rich stretch of DNA located in the intergenic region between the baiA2 and baiF genes from Eubacterium sp. strain VPI 12708 was amplified by polymerase chain reaction (PCR) and inserted in front of the Shine-Dalgarno (SD) sequences of three inefficiently-expressed Eubacterium sp. strain VPI 12708 genes cloned in Escherichia coli plasmids. Insertion of this A/T-rich cassette increased gene expression in all cases tested. Deletion of part of the A/T-rich region from a baiF clone in pUC19 resulted in decreased gene expression. Synthesis of specific mRNA was increased with addition of the A/T-rich cassette to constructs containing the baiC gene from Eubacterium sp. strain VPI 12708, but mRNA synthesis was not significantly changed in cells containing plasmid constructs with the baiF and baiG genes. Enhanced translation resulting from a decrease in mRNA secondary structure in the ribosome binding site region is discussed as a possible reason for increased gene expression with the A/T-rich cassette. PMID- 10367281 TI - Fundamentals of flow cytometry. AB - Flow cytometers are instruments that are used primarily to measure the physical and biochemical characteristics of biological particles. This technology is used to perform measurements on whole cells as well as prepared cellular constituents, such as nuclei and organelles. Flow cytometers are investigative tools for a broad range of scientific disciplines because they make measurements on thousands of individual cells/particles in a matter of seconds. This is a unique advantage relative to other detection instruments that provide bulk particle measurements. Flow cytometry is a complex and highly technical field; therefore, a basic understanding of the technology is essential for all users. The purpose of this article is to provide fundamental information about the instrumentation used for flow cytometry as well as the methods used to analyze and interpret data. This information will provide a foundation to use flow cytometry effectively as a research tool. PMID- 10367282 TI - Mechanisms and assessment of lectin-mediated mitogenesis. AB - The discovery of lectin-mediated mitogenesis by Nowell in 1960 stimulated interest in the properties of lectins while advancing knowledge of immunology. Although some lectins are polyclonal activators both in vitro and in vivo, others may display a broad range of activities toward human lymphocytes. Indeed, the same lectin (e.g., wheat germ agglutinin or Datura lectin) may be mitogenic, comitogenic, or antimitogenic, depending on the experimental conditions. An individual lectin may bind to several glycoproteins on the lymphocyte surface, resulting in interactions that may or may not be functionally relevant, and that may have opposing effects. Studies with lectins and with monoclonal antibodies (MAbs) have established that a surprisingly large variety of cell-surface molecules can influence the initiation and regulation of lymphocyte activation and proliferation. Interactions between lymphocytes and accessory cells are crucial; some signals are cell-mediated, but others depend on soluble cytokines. Mitogenic lectins presumably bind to the T-cell receptor complex and also promote a positive costimulatory signal leading to the synthesis of interleukin 2 and interleukin 2 receptors (IL-2R). Nonmitogenic, comitogenic, and antimitogenic lectin activities also probably act via accessory molecules involved in costimulation. Plant lectin-animal lymphocyte interactions presumably have no physiological significance, but it is suggested that the former mimics microbial superantigens, which may function in the colonization of host cells. Mitogenic stimulation of lymphocytes can be assessed in several ways. The standard technique measures [3H]-thymidine incorporation into DNA, but nonradioactive procedures are also available. PMID- 10367283 TI - Gene targeting of retinoid receptors. AB - Gene targeting in embryonic stem (ES) cells has been employed to investigate the role of the retinoid receptors and binding proteins both in the mouse as well as in embryocarcinoma cells. It is a powerful technique for the modification of the mouse genome. With more recent refinements in gene targeting technology, it is now possible to introduce more subtle mutations in the murine genome, as well as to investigate gene function in a tissue and temporally-restricted manner. It should also be possible to modify genes in diverse diploid cell lines, to generate diverse model systems for analysis of retinoid receptor function. In this article, some of the basic principles for gene targeting are described. PMID- 10367284 TI - Serology and urea breath test in the diagnosis of H. pylori infection. AB - Helicobacter pylori is a chronic infection that has the potential for causing and initiating serious gastric disease. Specific treatment can be successful in eradication of the infection but is currently complex which hampers essential patient compliance. Therefore, the accurate detection of H. pylori and, importantly, the post-treatment check for cure is vital in the effective management of this infection. This is especially true in cases of asymptomatic individuals. Serology is now a simple ELISA with a high degree of accuracy and has been shown to be useful as a screening tool prior to endoscopy in selected cases. The urea breath test, either using C13 or C14, is a sensitive test easily applied and is the test of choice for post-treatment check for cure. It is also the gold standard for the validation of serology in different populations. PMID- 10367285 TI - Use of NO donors in biological systems. AB - Owing to the increased interest in the biological roles of nitric oxide (NO) the use of NO donors is a desired method of delivering NO to the tissues of interest. This article gives an overview of the most commonly used classes of NO donors and their biotransformation to release NO. A major consideration when choosing an NO donor is the preparation and handling of the compounds. A method has been outlined for the preparation of S-nitrosothiols which eliminates the problem of the overall instability of these compounds both as a solid and in solution. The main aim of this article is to outline the methods used in assessing the ability of NO donors to elicit a biological response in vitro in particular relaxation of vascular smooth muscle and inhibition of platelet aggregation. In addition a method is described for assessing the toxicological potential of NO donors in vitro. PMID- 10367286 TI - A cloning vector for efficient generation of cholera toxin B gene fusions for epitope screening. AB - Gene fusion proteins with epitopes attached to the amino end of cholera toxin B subunit (CTB) are useful to raise immunological responses. We describe a cloning vector, designated pCTBtet, carrying a tetracycline resistance gene (TetR) between the leader peptide and mature CTB. Removal of TetR to insert oligonucleotides encoding fusion epitopes allowed for screening of tetracycline sensitive clones. Restoration of the correct CTB reading phase was subsequently used to choose gene fusion candidate colonies. The use of pCTBtet permitted the rapid construction of 8 fusion proteins carrying 9-24 aa from Salmonella typhi OmpC and 6 hybrids with 7-31 aa from Escherichia coli colonization factor CFAI. PMID- 10367288 TI - Self-injurious behaviour: a kindling phenomenon? AB - Self-injurious behaviour continues to be a drain on the resources for rehabilitation of the children with developmental disabilities. Pathogenesis and therapy seems fragmentary, with virtually every major neurotransmitter system being identified as the putative substrate for self-injurious behaviour. This phenomenon as it cuts across the diagnostic boundaries, although it is suggestive of a heterogeneous neurochemical basis, should call for the exploration of the biological event preceding the neurochemical cascade resulting in the behaviour. The authors argue that kindling is the preceding neurophysiological event resulting in self-injurious behaviour and, thus, can be effectively prevented pharmacologically. PMID- 10367287 TI - [Electroencephalography of the premature and term newborn. Maturational aspects and glossary]. AB - From the first publication of C. Dreyfus-Brisac and N. Monod, a strong tradition combined with tremendous development of neonatal EEG has taken place in France. After 3 years of collaborative work, 12 clinical neurophysiologists trained at the Port-Royal medical school in Paris detail in this paper the currently available neonatal EEG recording techniques. They have synthesized the criteria of maturational state analysis and have defined the normal and pathological neonatal EEG patterns, including descriptions already present in the French as well as the English literature. In this review one may find a complete description of neonatal EEG patterns according to the states of vigilance and to gestational age. Furthermore, definitions of all normal and pathological patterns are provided in a glossary. Both chapters are illustrated by numerous figures. This detailed terminology in neonatal EEG should allow a better homogeneity in EEG reports, and could lead to multicentric studies on normal, unusual or pathological patterns, according to etiology. Although based on analogic EEG data, this work can equally be applied to digitized EEG tracings. PMID- 10367289 TI - Real-time continuous visual biofeedback in the treatment of speech breathing disorders following childhood traumatic brain injury: report of one case. AB - The efficacy of traditional and physiological biofeedback methods for modifying abnormal speech breathing patterns was investigated in a child with persistent dysarthria following severe traumatic brain injury (TBI). An A-B-A-B single subject experimental research design was utilized to provide the subject with two exclusive periods of therapy for speech breathing, based on traditional therapy techniques and physiological biofeedback methods, respectively. Traditional therapy techniques included establishing optimal posture for speech breathing, explanation of the movement of the respiratory muscles, and a hierarchy of non speech and speech tasks focusing on establishing an appropriate level of sub glottal air pressure, and improving the subject's control of inhalation and exhalation. The biofeedback phase of therapy utilized variable inductance plethysmography (or Respitrace) to provide real-time, continuous visual biofeedback of ribcage circumference during breathing. As in traditional therapy, a hierarchy of non-speech and speech tasks were devised to improve the subject's control of his respiratory pattern. Throughout the project, the subject's respiratory support for speech was assessed both instrumentally and perceptually. Instrumental assessment included kinematic and spirometric measures, and perceptual assessment included the Frenchay Dysarthria Assessment, Assessment of Intelligibility of Dysarthric Speech, and analysis of a speech sample. The results of the study demonstrated that real-time continuous visual biofeedback techniques for modifying speech breathing patterns were not only effective, but superior to the traditional therapy techniques for modifying abnormal speech breathing patterns in a child with persistent dysarthria following severe TBI. These results show that physiological biofeedback techniques are potentially useful clinical tools for the remediation of speech breathing impairment in the paediatric dysarthric population. PMID- 10367290 TI - Outcome measure for paediatric rehabilitation: use of the Functional Independence Measure for children (WeeFIM). A pilot study in Chinese children with neurodevelopmental disabilities. AB - PURPOSE: To study the use of Functional Independence Measure for children (WeeFIM) in monitoring neurorehabilitation programmes for children with neurodevelopmental disabilities. METHODS: The neurorehabilitation team of the Children's Habilitation Institute of the Duchess of Kent Children's Hospital were trained to administer the WeeFIM. The WeeFIM was administered to children with various neuro-developmental impairment groups undergoing neurorehabilitation programmes in the hospital inpatient and also outpatient setting. The WeeFIM was scored on hospital admission and prior to discharge for those admitted for the rehabilitation programme. The WeeFIM profile was then monitored half yearly. The pilot study used WeeFIM in assessing 104 children with different medical disease categories. The disease or impairment categories included very low birth weight babies (n = 44), cerebral palsy (n = 19), Down's syndrome (n = 9), pervasive developmental disorder (n = 11), Duchenne Muscular Dystrophy (n = 18), and others (n = 3). RESULTS: WeeFim could be used to measure disability, monitor progress, enhance communication, measure the effectiveness of treatment, and document the benefits of rehabilitation intervention. It also served as a networking of neurorehabilitation programmes for different impairment categories in a continuum of settings: hospital, community, school and at home. WeeFIM was found to be a quick and reliable functional assessment instrument in this rehabilitation facility. CONCLUSIONS: WeeFIM could be used to assist neurorehabilitation clinicians in the selection of short term realistic goals and long term rehabilitation strategies for children with various neurodevelopmental disabilities, and the subsequent progress of the children could be monitored objectively. PMID- 10367291 TI - The prevalence of attentional problems and the effect of methylphenidate in children with myelomenigocele. AB - The prevalence of attentional problems, and the effect of methylphenidate was evaluated in a clinic population of children with myelomeningocele. Families of 79 children between the ages of 6 and 15 years were screened for the presence of attention problems in their children, using Conners' questionnaires for parents and teachers, and/or the DSM-IV checklist. Thirty-nine per cent of the children exhibited attention problems, primarily without hyperactivity. Fourteen children with attentional problems were enrolled in a double-blind placebo-controlled trial methylphenidate. Response to methylphenidate was assessed with Conners' questionnaires, Conners' Continuous Performance Test, and a battery of selected neuropsychological tests. No statistically significant response was measured for the group while on methylphenidate. Four children were clinical responders to methylphenidate. The prevalence of attentional problems in children with myelomeningocele is high, and effective medication therapy needs to be studied further. PMID- 10367292 TI - Effects of angiogenic growth factors on endothelium-derived prostacyclin production by ovine uterine and placental arteries. AB - Uteroplacental and fetoplacental arteries produce substantial amounts of prostacyclin (PGI2). Because angiogenic growth factors such as basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), and epidermal growth factor (EGF) are increased in pregnancy, we hypothesized that treatment of uterine and fetoplacental arteries with bFGF, VEGF, and EGF would further enhance the pregnancy-induced increase in PGI2 production. Duplicate uterine (UA) and fetoplacental (PA) artery (primary branch off of the umbilical cord = pPA; cotyledonary or tertiary = tPA) explants from seven late gestation sheep were placed in tissue culture (RPMI; 37 degrees C) for 24 h alone or with (1-100 ng/mL) bFGF, VEGF, or EGF. To evaluate the endothelial contribution to basal and stimulated PGI2 production and to determine whether it is de novo, arteries with and without endothelium from three additional late gestation ewes, tissues were incubated in the absence or presence of growth factors with or without meclofenamate (1 microM). The stable metabolite of PGI2, 6-keto-PGF1 alpha, was measured in culture media and expressed as ng/mg wet wt 24 h. PGI2 production by UA increased (p < 0.05) from 5.43 +/- 0.26 at control to 8.93 +/- 0.99 with 100 ng/mL bFGF. Although VEGF produced a similar response, EGF did not increase PGI2 production in UA. In pPA, 100 ng/mL bFGF induced a 2.2-fold increase (p < 0.01) in PGI2 production from 1.94 +/- 0.14 to 4.20 +/- 0.31; VEGF and EGF were without effect. In tPA, 50 and 100 ng/mL bFGF increased PGI2 production from 1.98 +/- 0.14 to 3.5 +/- 1.05 and 3.96 +/- 0.46 (p < 0.02). In tPA, VEGF did not increase PGI2 production; however, 10, 50, and 100 ng/mL EGF, enhanced (p < 0.03) PGI2 production from 1.98 +/- 0.14 to 3.39 +/- 0.62, 3.62 +/- 0.26, and 2.93 +/- 0.20. Endothelium removal and meclofenamate treatment caused a 90% and 100% decrease, respectively, in basal PGI2 production, with no recovery after treatment with growth factors. We conclude that PGI2 production is augmented by bFGF in UA, pPA and tPA, by VEGF in UA, and by EGF in tPA during ovine pregnancy. Basal and stimulated PGI2 secretion is endothelium-derived via de novo synthesis. bFGF, VEGF, and EGF, in addition to angiogenesis, may modulate PGI2 production, further enhancing blood flow to the growing uteroplacental bed. PMID- 10367293 TI - PGE receptor characteristics on porcine luteal cells during the estrous cycle and early pregnancy. AB - This study examined the affinities and concentrations of prostaglandin E (PGE) receptors on porcine luteal cells during the estrous cycle and early pregnancy. Corpora lutea (CL) were obtained from nonpregnant gilts at days 9 (n = 4), 12 (n = 3), and 14 (n = 6); three gilts possessed red, vascular CL and three gilts had white nonvascular CL) of the estrous cycle, and days 9 (n = 4), 12 (n = 3), 14 (n = 5), and 30 (n = 5) of pregnancy. The CL were dissociated enzymatically to disperse single cells and the red blood cells were removed by elutriation. The luteal cells were assayed for specific PGE binding by displacement analysis with use of [3H] PGE2 and varying concentrations of unlabeled PGE2. The specific binding of [3H] PGE2 to luteal cells decreased (p < 0.05) from days 9 to 14 of the estrous cycle, but only decreased (p < 0.05) from days 9 to 12 of pregnancy. Specific binding was higher (p < 0.05) on day 14 of pregnancy than the comparable stage of the estrous cycle. The affinities of PGE receptors decreased (p < 0.05) only on the luteal cells dissociated from red, vascular CL of day 14 nonpregnant gilts compared with those of other days of the estrous cycle and pregnancy. The number of PGE receptors on porcine luteal cells was similar (p > 0.05) in pregnant and nonpregnant gilts, but decreased (p < 0.05) on days 12-14 postestrus. During early pregnancy, it was evident that high affinity PGE receptors are sustained on porcine luteal cells; however, the role of the PGE receptors in maternal recognition of pregnancy remains speculative. PMID- 10367294 TI - Opiate, cannabinoid, and eicosanoid signaling converges on common intracellular pathways nitric oxide coupling. AB - Scientific fields as they emerge initially appear to be unrelated to other projects even if they are in a similar area of interest. This is especially true in the case of opiate, cannabinoid, and eicosanoid signaling processes. In this limited speculative review, we attempt to examine aspects of their intracellular cascading signaling systems for their commonalities. We find intracellular calcium mobilization, nuclear factor kappa B involvement, adenylate cyclase activity, and, finally, constitutive nitric oxide release to be converging points for these signaling processes, occurring by separate and distinct receptor mediated effector systems. Phosphokinase C, mitogen activated protein kinase, and cytosolic phospholipase A2 also represent points of common impact. In this regard, aspirin also appears to be involved in an aspect of this signaling convergence. We conclude that many of the physiological observations regarding the actions of these signaling molecules, for example, immunosuppression, neurotransmission, vasodilation, cellular adherence, and cytotoxicity, can now be understood by considering their converging biochemical cascades. PMID- 10367295 TI - Platelet-activating factor and eicosanoids are mediators of local and systemic changes induced by immune-complexes in mice. AB - A passive Arthus reaction (AR) induced in the peritoneal cavity of mice was followed by increased local vascular permeability and haemoconcentration. The intensity of the increased vasopermeability was higher in BALB/c compared with C3H/HePas mice despite the latter being 10 times more sensitive to platelet activating factor (PAF). C3H/HePas mice however, exhibited higher levels of haemoconcentration and shock-like symptoms. Both events were inhibited by the PAF antagonist, WEB 2170. Indomethacin reduced both pathological events whereas L663,536, that inhibits leukotrienes synthesis reduced haemoconcentration but only in BALB/c mice. PAF was released into the peritoneal cavity, peak release being at 10 min after induction of AR. Prostaglandin E2 (PGE2), thromboxane B2 (TXB2), leukotriene B4 (LTB4), and leukotriene C4/D4 (LTC4/D4) were also released at this time. Similar levels of PAF and eicosanoids were found in BALB/c and C3H/HePas mice except for LTB4, which was higher in C3H/HePas. It is concluded that PAF and eicosanoids are mediators of local and systemic changes induced by immune complexes in the peritoneal cavity of mice. PMID- 10367297 TI - [Epidemiology and public health. Pursuing the clarification effort]. PMID- 10367296 TI - Nitroprusside stimulates contractility and the synthesis of 14C-acetylated PAF like substances in estrogen primed-mouse uterine horns. AB - We investigated the effect of sodium nitroprusside (SNP), a donor of nitric oxide, on the formation of platelet-activating factor (PAF) and uterine contractility in mouse uterine horns from mice treated with estrogen. Because the major pathway of PAF synthesis is the remodeling pathway in uterine tissue, we evaluated the incorporation of 14C-acetate into PAF-like molecules. Our results showed that SNP (100-300 mumol/L) caused a transient increase in the synthesis of PAF, which remained cell-associated. The addition of SNP (100-300 mumol/L) to a mouse uterine horn in an isolated organ bath preparation evoked a transient increase in contractility, which was inhibited by hemoglobin (2 micrograms/mL), a nitric oxide scavenger, but not by methylene blue (10 mumol/L), a guanylate cyclase inhibitor. The pharmacological characteristics of the contractions evoked by SNP resembled those evoked after mast cell activation, in that they were blocked by ritodrine (a beta 2 adrenergic agonist, 0.1 mumol/L); indomethacin (a cyclooxygenase inhibitor, 10 mumol/L); ketotifen (a mast cell stabilizer, 1.0 mumol/L); cromolyn sodium (a mast cell stabilizer, 100 mumol/L); pyrilamine (an H1 antagonist, 10 mumol/L); and ketanserine (5HT2 antagonist, 0.1 mumol/L). These data demonstrate that nitric oxide generated from SNP stimulated the synthesis of PAF and evoked contractility in uterine horns from mice treated with estrogen. This result suggests the possibility that these tissue conditions might be favorable for the generation of peroxynitrites, possible mediators of both effects. It is also shown that the contractility evoked by the addition of SNP was not due to production of PAF, because its antagonist, WEB 2086 (10-30 mumol/L, a concentration that blocked contractions evoked by PAF 1 nmol/L), had no effect on the SNP-evoked contractions. PMID- 10367298 TI - [Factors associated with survival in Kaposi's sarcoma in patients infected with the human immunodeficiency virus. Clinical Epidemiological Group for AIDS in Aquitaine]. AB - BACKGROUND: To study behavioral risk factors of Kaposi's sarcoma (KS) among HIV infected homosexuals in Bordeaux, southwest France. METHODS: A case-control study was performed within the Aquitaine Cohort. Cases of KS surviving in 1995 and homosexuals were systematically enrolled. For each case, two controls were selected among homosexuals surviving in the cohort. Cases and controls were matched on year of diagnosis of HIV infection. Data collection was based on a self administered questionnaire focusing on use of recreational drugs, detailed sexual practices and sexually transmitted diseases in the year preceeding the diagnosis of HIV infection, in the year after the HIV diagnosis and in the year preceeding the diagnosis of KS (or an equivalent period of time for controls). RESULTS: Twelve cases were matched to 2 controls, 15 cases to one control and 13 cases remained unmatched. Matched analysis identified an association between KS and regular sexual partner (odds ratio = 0.07; 95% confidence interval: 0.01-0.52 and p < 0.001) and active and passive oro-anal intercourse before HIV diagnosis and before KS diagnosis (p = 0.01). In the unmatched analysis including all cases, we found an association between KS and the overall number of sexual partners (p < 0.03) for all periods of interest. CONCLUSIONS: This case-control study identified sexual practices in favor of a sexually transmitted agent of KS. PMID- 10367299 TI - Can Flemish women in semi-rural areas be motivated to attend organized breast cancer screening? AB - BACKGROUND: The implementation of organized breast cancer screening in Flanders was prepared by means of pilot projects within a multicenter study. In the semi rural district of Kontich (Province of Antwerp, Flanders) a pilot project was performed using a mobile screening unit. Compared to international standards, the attendance rate for this pilot project (i.e. 34%) was low. Non-organized screening, which already exists in Flanders, at least partly explains this low attendance rate for the organized screening. The main purpose of our study was to investigate the experience of the pilot target group with respect to the organized breast cancer screening in the district of Kontich, in order to maximize the conditions for a high attendance rate in the organized breast cancer screening programme throughout Flanders. METHODS: With a random numbers procedure, performed by the computer, 500 women were selected among those who were invited to the first screening round of the breast cancer screening programme in the district of Kontich (n = 6,897). These 500 randomly selected women were asked to cooperate with a face-to-face interview. The questionnaire used dealt with the different aspects of the organized mammographic screening which were expected to influence the decision to attend. RESULTS: There were 348 women who responded to the questionnaire (69.6%): 138 of them were attenders and 210 were non-attenders at the organized breast cancer screening. Attenders and non-attenders at the organized breast cancer screening in the district of Kontich had different views about various aspects of the screening programme. The percentages of those who thought that an item was important or very important to them, were for the 138 attenders and the 210 non-attenders respectively: "to receive a personal invitation letter": 90.6 vs. 48.1% (p < 0.05); "a preliminary visit to the GP": 9.4 vs. 34.3% (p < 0.05); "possibility of examination outside business hours": 15.9 vs. 30.0% (p < 0.05). CONCLUSIONS: Although the putting into action of a mobile unit in the semi-rural area of the district of Kontich was productive, the attendance rate was still too low compared to international standards. To increase the attendance rate, the following interventions should be considered: devising the personal invitation letter in a more attractive way, activating and stimulating the important motivational role of the GP in persuading women to attend the organized screening programme and offering the invited population the possibility to have a mammographic examination performed outside business hours. Appropriate measures are being explored. PMID- 10367300 TI - [Comparison of two types of behavior and attitude surveys on alcohol, tobacco and illegal drug use]. AB - BACKGROUND: The aim of this study was to compare the results of two types of surveys frequently used in France to monitor alcohol, tobacco and illegal drug consumptions. METHODS: Data from a random sample phone survey (1993 18-75 year old French adults in November and December 1995), a quota sample face-to-face survey (1000 18-75 year old French adults in November 1995) and other sources (sales statistics and other surveys) were compared. RESULTS: Results from the two types of surveys were similar for alcohol and tobacco consumption prevalence. Concerning smoking prevalence, the random survey gave an estimate of 35.5% for the smokers ratio versus 35.2% with the quota sample survey. The daily alcohol consumer percentage was 20.9% and 22.1% respectively with the random and the quota sample surveys. Differences were observed for attitudes and illegal substances consumption: the random sample phone survey estimated at 15.5% people using drug during their life versus 13.9% for the quota sample face-to-face survey. CONCLUSIONS: Quota sampling and face-to-face surveys can be used to monitor alcohol, tobacco consumption and to a lesser extent, drug consumption trends, especially by repeated surveys, instead of random sample phone survey which are more time and money consuming. PMID- 10367301 TI - [Reliability and validity of the French version of the first part of the Appropriateness Evaluation Protocol (AEPf): pertinent criteria of hospitalization stay]. AB - BACKGROUND: The Appropriateness Evaluation Protocol (AEP) is a tool used to identify hospitalization days that are appropriate with respect to simple technical criteria, and the reasons for hospitalization days that do not meet these criteria. The goal of the study was to validate the clinical criteria of AEPf, required for identifying the reasons which explain these inappropriate hospital days. METHODS: The validity of a French version of criteria of appropriateness (AEPf) was assessed in a sample of 502 hospital days in medical and surgical wards of six French teaching hospitals. The reliability of the AEPf based conclusions was studied by the measure of the concordance between two independent measures of AEPf for the same hospital day. Validity of AEPf was studied by the measure of the concordance between AEPf-based conclusions and the judgments of physicians using implicit criteria to assess the necessity of the hospital day studied. The degree of concordance was estimated by the Kappa coefficient. RESULTS: AEPf reproducibility was high (Kappa coefficient: 0.81; 95% confidence interval (95% CI): 0.76-0.87). Validity was also high (Kappa coefficient: 0.61; 95% CI: 0.53-0.68). CONCLUSION: The French version of the AEP was thus shown to be both reliable and valid to identify hospitalisation days appropriate with respect to hospital's technical equipment and specific resources. However, the reproducibility of the assessment of reasons for hospital days that did not meet AEP criteria will require a validation. PMID- 10367302 TI - [Home and outside home food complementarity in Bamako (Mali): nutritional and economic aspects. What is the rationality behind consumers' choices?]. AB - BACKGROUND: Great modifications in social and family relationships and life style come from rapid urbanisation in developing countries. Various types of malnutrition coexist in these towns. Food consumption outside the home is more and more common. This in turn encourages rapid growth in the food informal economic sector which must be taken into account in food and nutrition policy and planning. The aim of this study was to analyse the characteristics and complementarities between home and outside the home food consumption of different kinds of individuals coming from different kinds of families in Bamako, in terms of expenditures and aspects of food and nutritional intakes. METHODS: 366 individuals from 74 families were interviewed. They were chosen according to defined criteria in three districts of Bamako of high, middle and poor socio economic level. RESULTS: At home the daily food expenditure is 2.27 and 3.79 times greater per individual in rich than in middle income and poor families respectively. Animal proteins are respectively 41%, 19% and 9% of daily protein intake. Energy from lipids is 20 to 30% in rich and middle families. In poor ones it is only 15% which is the lower limit of nutritional recommendations. Moreover, contrary to proteins and carbohydrates, the cost of lipids seems, almost incompressible. Almost everybody eats out of home food, particularly children. Its cost, on an energy basis, is higher than home food. The expense is 19 to 27% of the family food budget. It appears necessary to the satisfaction of nutritional requirements in middle income and poor families. Despite various costs, whatever be the socio-economic level, energy intakes coming from that food are equivalent in absolute terms among various kinds of individuals: children, men and women. CONCLUSION: Families had to adapt their food strategies after the 1994 Franc CFA devaluation. Various hypotheses are presented, linked to intra family relationships and, within poor families, to insertion in the street food economic sector, in order to understand, the logic of food choices. Such an analysis, where health, nutritional, economic, social and cultural aspects of food are taken into account, allows some concrete orientations for urban food and nutrition policy. PMID- 10367303 TI - [Retrospective estimation methods of pesticide occupational exposure]. AB - The extensively growing use of pesticides in the last decades has led to great improvements in agriculture but also to threats for human health. Studying long term effects to assess individual exposures encounters difficulties in exposure assessment. This article summarizes retrospective methods for assessing occupational exposures to pesticides currently used in epidemiological studies. Questionnaires, environmental and biological monitoring constitute direct assessment of exposure, in an individual approach. But the validity of questionnaires is often poor and the metrology rarely reflects past exposures. Some indirect measures have been used together with job exposure matrices. To establish dose-response relationships is important to quantify the risk, but needs accurate exposure assessment based on quantitative indexes. PMID- 10367304 TI - [Evaluation of patient satisfaction in health facilities. Review of the literature]. AB - Assessment of hospital patient satisfaction is henceforth a statutory obligation in France. The purpose of this study was to define the main settings of patient satisfaction surveys and to describe methods for assessing patient satisfaction, through a literature review. Published works have mainly been based on experts opinions; few studies have used appropriate methodology. On the whole, their authors consider that patient satisfaction assessment should provide a useful feedback to the quality improvement system of hospitals. Surveys should associate qualitative and quantitative techniques and analysis of patient complaints and letters. Patient satisfaction surveys should not be separated from theoretical work on the foundations of this concept and its measurement. PMID- 10367305 TI - [Paradigms and pragmatism]. PMID- 10367306 TI - [Misuse of genetic tests by insurance companies]. PMID- 10367307 TI - [Double-blind placebo-controlled European trial for systolic hypertension--and prevention of dementia]. PMID- 10367309 TI - [Epidemiology of childhood asthma in the department of Calvados]. AB - A survey was conducted in 1707 sixth grade school children in the Calvados department of France. A self-administered questionnaire was filled out by the children in the presence of a school nurse. The cumulative prevalence of asthma was 14.9%. There was no significant difference between children living in urban or rural areas. There was however a significant difference by sex: 18% of the boys had asthma and 11% of the girls. The cumulative prevalence of wheezing was 25.4% (current prevalence 12.9%). The current prevalence of dry nocturnal cough, respiratory infections excluded, was 33.3%; that of exercise-induced asthma, 27%. Severity was evaluated on the basis of the number of wheezing episodes since the beginning of the school year (> 3 episodes: 4.5%), the number of awakenings at night per week (several per week: 2.2%), and aggravations severe enough to bother speech (4.3%). The rate of missed school days was 18.5% and that of asthma related hospitalization 1.8%. One asthmatic child out of 4 had undergone pulmonary function tests and 1 out of 2 had a specific treatment for asthma. The high prevalence of childhood asthma in Calvados, with high morbidity, a significant number of missed school days, the exceptional nature of satisfactory pulmonary function testing, and the inconsistency of specific treatments, emphasizes the need for educational programs for parents and their family and improved physician training. PMID- 10367310 TI - [Intrathoracic coelomic cysts]. AB - Intrathoracic coelomic cysts are benign embryonic tumors with a mesothelial lining. The aim of this work was to review possible localizations (pleuropericardic and other), the remaining surgical indications, and the current situation of minimally invasive techniques. We reviewed retrospectively, 28 cases of intrathoracic coelomic cysts in 12 men and 16 women, mean age 44 years. We recorded the cyst localization, clinical signs, indication for surgery, access routes used, and outcome. Twenty-one cysts were pleuropericardial cysts and 7 were ectopic mediastinal cysts. In all 7 of the ectopic mediastinal cysts and 4 of the pleuropericardial cysts surgery was indicated for diagnosis; for the other pleuropericardial cysts the indication was based on clinical signs (n = 4), large volume (n = 4), progressing volume (n = 7), no apparent reason (n = 1) and association with surgery for pneumothorax (n = 1). Assess was by mediastinoscopy (n = 1), mediastinotomy (n = 1), sub-xyphoid route (n = 1), thoracotomy (n = 18), and videothoracoscopy (n = 7). Long-term outcomes (mean follow-up 4 years 4 months) were good with no recurrences. Postoperative sequelae were observed in 6 cases after thoracotomy and in 1 case after videothoracoscopy. In summary, pleuropericardial cysts warrant surveillance without surgery unless their volume increases or clinical signs develop. Ectopic mediastinal cysts usually require surgery for diagnosis. It would appear advisable to prefer videothoracoscopy which allows diagnosis and excision of pleuropericardial cysts. Minimal thoracotomy may be helpful for ectopic mediastinal cysts. PMID- 10367311 TI - [Malignant small-cell thoracic pulmonary tumor (Askin tumor)]. AB - We report 4 cases of malignant thoraco-pulmonary small-cell tumors (Askin tumor). Only two cases were operated. We emphasize the difficult histological diagnosis and demonstrate the importance of complete removal for survival. Prognosis remains poor. PMID- 10367312 TI - [Fire-eater's lung: report of six cases]. AB - Lung damage due to inhalation of inflammable products, especially Kerdane, is mainly observed in young subjects. Lung damage is rarely severe and the diagnosis is facilitated by the notion of inhalation. These conditions regress favorably in a few days. We report six cases. PMID- 10367313 TI - [Acute respiratory distress in a patient with sickle-cell anemia]. AB - The acute chest syndrome is a frequent complications of sickle-cell disease characterized by chest pain, fever, and new infiltrate on chest x-ray image. Pathophysiologic factors appear to be multifactorial and better known. We report the case of a 28-year-old woman with homozygous sickle cell anemia who developed acute chest syndrome probably secondary to fat embolism. PMID- 10367314 TI - [A rare cause of hemoptysis]. AB - The existence of a bronchial foreign body is an unusual cause of haemoptysis. We observed a sixty two year-old women who presented several medium-abundance haemoptysis. They were associated with a systematic alveolar-interstitial radiological picture of the ventral upper right lobe. A right upper lobectomy showed that an old bronchial foreign body (piece of bone) was responsible for the systematic intra-alveolar bleeding. Though most of the breathed foreign bodies are expressed into immediate symptoms, some of them remained undiagnosed and may be responsible for haemoptysis, infectious complications, atelectasis and for bronchiectasis. Their extraction through endoscopy or most often surgery is necessary for a proper recovery. In spite of histopathological differences between foreign bodies, broncholithiasis and lung tumor the diagnosis may be difficult clinically and on radiology. PMID- 10367316 TI - [Mercury pulmonary embolism. Two case reports]. AB - Mercury pulmonary embolism following intentional or accidental injection of metallic mercury are uncommon. Generally, there are few clinical signs (acute pneumopathy in 50% of the cases). Chest x-ray shows multiple and bilateral point opacities with a metallic density. We present two cases of mercury pulmonary embolism after intentional intravenous mercury injection (one attempted suicide and one HIV+ drug addict). PMID- 10367315 TI - [Recurrent bilateral pleurisy in an 80-year-old man]. AB - An 80-year-old man was admitted with recurrent asphyxiating pleurisy, first attributed to heart failure. During the recurrent episodes, the patient presented fever, signs of inflammation, no signs of heart failure, and subnormal cardiac function, prompting further investigations which disclosed that the patient was a homozygous carrier of the severe type of periodic disease mutation. The patient's age at symptom onset and the clinical features of this case of periodic disease are exceptional. These points emphasize the usefulness of available genetic tests in difficult diagnostic cases. It also reflects current difficulties in trying to establish correlations between genotype and phenotype in periodic disease. PMID- 10367317 TI - [An anterior mediastinal mass]. AB - A 50-year-old man developed a bronchogenic cyst complicated by hemorrhage. A complete radiographic chest work-up provided a reliable diagnostic approach. Bronchogenic cysts are usually asymptomatic incidental discoveries. Chest ultrasonography confirms the cystic nature of the mediastinal mass. Computed tomography scan and especially magnetic resonance imaging further support the diagnosis and are helpful for guiding surgery. Surgery is required because of the unpredictable risk of hemorrhage, infection or enlargement. PMID- 10367318 TI - [Positive angiogram sign: value in the diagnosis of oil-aspiration pneumonia in an elderly patient]. PMID- 10367319 TI - [How phase III therapeutic trials in multiple sclerosis should be evaluated: advances and challenges]. PMID- 10367320 TI - [Interferon beta-1a (Avonex TM): clinical and MRI impacts]. AB - This article presents critical aspects of the pivotal phase III study that led to approval of interferon beta-1a (Avonex) as treatment for relapsing-remitting multiple sclerosis (MS). An update on data from the pivotal study or other open studies, further demonstrating the practical clinical impact of Avonex, are also presented. The purpose of the pivotal study was to determine whether Avonex could slow the progressive irreversible neurological disability of relapsing-MS. 301 patients were randomised into a double-blind placebo-controlled, multicentric trial of Avonex. Avonex 6.0 MUI (30 micrograms) was administered by intramuscular injection weekly. The primary outcome variable was time to sustained physical disability at 6 months progression of at least 1.0 point on the EDSS. Avonex produced a significant delay in time to sustained EDSS progression (p = 0.02). The Kaplan-Meier estimate of the proportion of patients progressing by the end of 104 weeks was 34.9 p. 100 in the placebo group and 21.9 p. 100 in the Avonex treated group, representing a 37 p. 100 reduction in the risk of accumulating disability. Further analysis showed that the clinical efficacy related to disability did not depend on the definition of disability progression. Significantly fewer Avonex recipients progressed to EDSS milestones of 4.0 or 6.0. The exacerbation frequency was decreased by 32 p. 100 in the Avonex group. Patients treated with Avonex had also a significantly lower number and volume of gadolinium-enhanced lesions on MRI. There was no major adverse events related to treatment. Injection site reactions were rare and no reports of skin necrosis have been recorded. There was no significant and clinically relevant biological disturbances due to Avonex. PMID- 10367321 TI - [Effect of weekly intramuscular interferon beta-1a on MRI lesions in patients with relapsing multiple sclerosis]. AB - We compared the number of new gadolinium-enhancing (Gd+) and T2-weighted (T2W) lesions on 6-monthly MRI scans before and after initiating weekly intramuscular injections of interferon beta-1a (Avonex) 30 mcg. The mean number of new focal Gd+ lesions detected during the 6 monthly on-treatment scanning sessions was 58 per cent less per MRI scan and 62.5 per cent less per patient than during the 6 monthly pre-treatment scanning sessions (p = 0.016, Wilcoxon signed rank test). The results are similar for new focal Gd+ and T2W lesions. The reduction in disease activity detected by monthly Gd+ enhanced MRI scans after initiating weekly intramuscular interferon beta-1a 30 mcg is consistent with benefits observed with thrice weekly subcutaneous interferon beta-1a 10 and 30 mcg and alternate day subcutaneous interferon beta-1b 8.0 MIU. PMID- 10367322 TI - [Beta interferon in multiple sclerosis: pharmacological differences]. AB - Interferons beta (INF) would be a good recombinant therapeutic choice for the management of relapsing remitting multiple sclerosis (RRMS). Three forms are approved in various countries including one IFN beta-1b (Betaferon) and two IFN beta-1a (Avonex and Ribif). These agents are apparently similar although they are not identical. Differences may be of clinical relevance. Important differences are described here in terms of pharmacokinetics, the spectrum of side effects and molecular chemistry. The clinical consequences on the benefit/risk ratio are discussed. This review recalls the difficulty in developing such compounds and in reaching marketing approval. An improvement in the acceptability and efficiency of IFN beta could by obtained when analyzing basic research data. PMID- 10367323 TI - Autosomal dominant spinocerebellar degenerations. Clinical, pathological, and genetic correlations. AB - Historical review on hereditary spinocerebellar degenerations (SCD) revealed that some CAG repeat diseases were formerly diagnosed under several different names because of their clinical and pathological heterogeneity. A genetic abnormality in these hereditary SCDs (often with expanded CAG repeat) corresponds to a definite prototypic combination of the principal lesions in the cerebellar, extrapyramidal, and oculomotor systems, which allows neuropathological differentiation between these entities. Variability of both clinical and pathological features is mainly related to the patient's age at onset, which is often correlated with the number of CAG repeat size. Several characteristics are suggestive of these SCD: preferential degeneration of specific systems, size reduction at cellular or tissue level not accompanied by glial reaction (simple atrophy or hypoplastic change) and presence of recently identified ubiquitin positive inclusions in neurons are characteristics of these SCDs. These features provide further insight into the phenotypic development of a genetic abnormality. PMID- 10367324 TI - [Enhanced adherence of endothelial cells to blood mononuclear cells in HAM/TSP]. AB - We investigated the adhesion of blood mononuclear cells (MNC) isolated from patients with HTLV-1-associated myelopathy (HAM/TSP). MNC from HAM/TSP patients were significantly more adherent to activated endothelial monolayers than MNC from non-HAM/TSP (controls and HTLV-1 carriers) subjects. Blocking studies demonstrated that the adhesion molecules VLA-4 (CD49d), ICAM-1 (CD54), and L selectin (CD62L) all contributed to increased binding. However, anti-ICAM-1 antibody was the most efficient in inhibiting binding HAM/TSP patients MNC to activated endothelial cells. Expression on MNC of molecules involved in adhesion was also studied by flow cytometry in HAM/TSP patients, HTLV-1 carriers, and healthy control subjects after two days culture without any mitogen. In HAM/TSP patients, L-selectin expression on CD4+ and CD8+ subsets was lower than in controls; interestingly, HAM/TSP patients had lower percentage of CD4+ subset expressing L-selectin than HTLV-1 carriers. The percentage of CD4+ and CD8+ cells expressing VLA-4 was found to be similar to controls in both HAM/TSP patients and HTLV-1 carriers. Following two days in culture without mitogen, the percentage of T cells expressing ICAM-1 increased in HAM/TSP and carriers, but not in controls. This study provides information regarding trans-endothelial migration of MNC across the blood brain barrier in HAM/TSP and suggests ICAM-1 and its counterpart molecule LAF-1 are involved in massive infiltration of lymphocytes observed in the spinal cord. PMID- 10367325 TI - [Postural balance following stroke: towards a disadvantage of the right brain damaged hemisphere]. AB - In the light of studies published in the last ten years, we have suspected a differential influence of the sides of hemispheric cerebral lesions on posture and balance. A study was aimed at verifying this hypothesis, the method of which being original because many possible confounding factors such as age, sex as well as topography and size of the brain lesion have been taken into account in the statistical analysis. Inclusion criteria were: right-handed patients, first stroke, no previous disease which might have affected balance. Their postural abilities (ranging from 0 to 36) were assessed 90 +/- 3 days after stroke onset on a clinical scale. This clinical assessment was used here because it could be easily performed in all patients, irrespective of the severity of their impairment. Lesion locations were determined using the Atlas by Talairach and Tournoux and the number of cerebral areas altered gave an estimation of the lesion size. The first fifty patients consecutively admitted to rehabilitation and responding to the inclusion criteria were thus examined (25 with a right and 25 with a left hemispheric lesion; 14 women and 36 men; mean age 57.2 = yrs). The main result was lower postural performances in right brain-damaged patients than in left brain damaged patients (21.5 vs 29.4; p = 0.01). Postural abilities were also inversely related to age (r = -0.28; p = 0.04), lesion size (r = -0.41; p = 0.003) and were lower in women than in men (22.1 vs 28.8; p = 0.02). This study therefore confirms the existence of a right hemispheric dominance for postural control. The existence of inverse correlation between postural abilities and the number of omitted targets in cancellation task on one hand (r = -0.63, p < 0.001), the ipsilesional bias in line bissection on the other hand (r = -0.36; p = 0.01), argued for a relationship between the main result of this study and the well known cerebral organization of spatial information processing, based on a right hemisphere dominance for spatial attention and/or representation. The 'postural neglect' concept is discussed. PMID- 10367326 TI - [Epilepsy and videogame: which physiopathological mechanisms to expect?]. AB - Video games may induce epileptic seizures in some subjects. Most of them have photosensitive epilepsy. The triggering factors are multiple: characteristics of the softwares, effects of the electronic screen and interactivity. The wide diffusion of the video games explain the large number of descriptions of videogame induced seizures. Historical aspects and an analysis of the underlying mechanisms of videogame induced seizures are presented. PMID- 10367327 TI - [Tibial muscular dystrophy. A rare form of distal myopathy]. AB - Tibial muscular dystrophy (TMD) is a dominantly inherited late onset distal leg myopathy only described in the Finnish population as yet. A similar disorder was described by Markesbery et al. in 1974 in one American family. Assignment of the TMD locus to chromosome 2q31 has been demonstrated (Haravuori et al., 1998). We recently described a French family with clinical and laboratory findings similar to TMD (de Seze et al., 1998). Molecular genetic results indicate that the distal myopathy in this family could be linked to the TMD locus confirming TMD exists outside the Finish population. This overview of TMD will allow to describe differential diagnoses such as other distal myopathies and scapuloperoneal syndromes. PMID- 10367328 TI - [Residual cerebellar ataxia following acute phenytoin intoxication]. AB - A 30-year-old man was given high doses of phenytoin together with 4 antituberculous drugs for a seizure associated with a probable brain tuberculoma. He developed hepatic toxicity and his serum phenytoin reached the high level of 298 mumol/l (therapeutic range 40-79 mumol/l). All drugs were stopped and the biological parameters returned progressively to normal over the next 15 days. However, he remained with a cerebellar axial syndrome and was still severely ataxic 2 months later. Brain CT and MRI showed mild cerebellar atrophy. This case and the few other published ones, together with some recent experimental data, show that high doses of phenytoin can be toxic to the cerebellar cortical cells. The rarity of similar cases, while millions of epileptics are under phenytoin treatment, would however suggest that individual susceptibility may play a role in this toxicity. PMID- 10367329 TI - Ependymoma of the fourth ventricle presenting with hemifacial spasm. Report of a case. AB - According to Gardner's hypothesis (1962) later confirmed by Jannetta (1982, 1985), hemifacial spasm can usually be related to a "vascular conflict" which takes place inside the cerebellopontine angle (CPA). Occasionally, the causative lesion can be identified as a mass encasing the facial nerve at its root exit zone (REZ) from the brain stem. The hemifacial spasm has been rarely reported in presence of a contralateral CPA mass ("false localising sign"). Hemifacial spasm in patients with masses in anatomical regions other than the CPA has to be considered exceptional. The case of an adult man harboring an ependymoma of the fourth ventricle whose only neurological sign was a left hemifacial spasm is reported. The rarity of such a condition prompted us to review the literature. Particular attention has been paid to the possible pathogenetic mechanisms and their therapeutic implications. PMID- 10367330 TI - [Pharmacological vigilance and pharmacological epidemiology: principles, definition, methods and current trends in neurology]. AB - It is now well established that only clinical trials performed before drug approval are not sufficient for a full modern pharmacological evaluation of drugs and treatments. The need of both pharmacovigilance and pharmacoepidemiology is underlined in order to evaluate drugs under real conditions. After a summary of methods used in pharmacoepidemiological trials (spontaneous reports, imputability assessment, cohorts, case control studies etc.), recent pharmacoepidemiological data useful for the neurologist are summarized: side effects of tacrine and vaccines, serotoninergic syndrome and side effects of new antiepileptic drugs. PMID- 10367331 TI - Breast motion and sports brassiere design. Implications for future research. AB - Exercise usually results in a large displacement of the breasts, often leading to breast pain. Although breast pain is a common concern of exercising females, little research has been conducted in the area of breast pain. It has been suggested that a cause of breast pain is excessive breast motion. As the female breast does not contain strong intrinsic structural support, this breast motion is difficult to reduce. It is suggested that the primary anatomical support for the breast is the Cooper's ligaments; however, their true functional properties are unknown. Because of the lack of internal breast support it has been suggested that the skin covering the breast may also act as a support structure for the breast, but this has not been quantified. In an attempt to reduce breast motion, external breast supports (brassieres) have been developed. This article discusses components of current sports brassieres with implications for future research required to improve brassiere design and performance. PMID- 10367332 TI - The zone diet and athletic performance. AB - The Zone diet is the latest eating regimen marketed to improve athletic performance by opposing traditional high carbohydrate sports diets. The 40/30/30 diet is centred primarily on protein intake (1.8 to 2.2 g/kg fat free mass; i.e. total bodyweight-fat weight) and promises a change in the body's insulin to glucagon ratio through its macronutrient alterations. Changes in the existing hormonal milieu are said to result in the production of more vasoactive eicosanoids, thus allowing greater oxygen delivery to exercising muscle. This favourable condition, known as the Zone, is anecdotally reported to benefit even the most elite endurance athletes. Applying the Zone's suggested protein needs and macronutrient distributions in practice, it is clear that it is a low carbohydrate diet by both relative and absolute standards, as well as calorie deficient by any standard. Reliable and abundant peer reviewed literature is in opposition to the suggestion that such a diet can support competitive athletic endeavours, much less improve them. The notion that a 40/30/30 diet can alter the pancreatic hormone response in favour of glucagon is also unfounded. The Zone is a mixed diet and not likely to affect pancreatic hormone release in the same way individual nutrients can. Although the postprandial insulin response is reduced when comparing a 40% with a 60% carbohydrate diet, it is still a sufficient stimulus to offset the lipolytic effects of glucagon. Many of the promised benefits of the Zone are based on selective information regarding hormonal influences on eicosanoid biology. Contradictory information is conveniently left out. The principle of vasodilating muscle arterioles by altering eicosanoid production is notably correct in theory. However, what little human evidence is available does not support any significant contribution of eicosanoids to active muscle vasodilation. In fact, the key eicosanoid reportedly produced in the Zone and responsible for improved muscle oxygenation is not found in skeletal muscle. Based on the best available scientific evidence, the Zone diet should be considered more ergolytic than ergogenic to performance. PMID- 10367333 TI - Anaemia and iron deficiency in athletes. Practical recommendations for treatment. AB - Trained athletes frequently experience low levels of blood haemoglobin (13 to 14 g/100ml in men and 12 g/100ml in women) plus low haematocrit and low ferritin levels. These parameters define the concept of 'sports anaemia'. Low iron levels may be due to mechanical haemolysis, intestinal bleeding, haematuria, sweating, low iron intake or poor intestinal absorption. The resulting decrease in blood gas transport and muscle enzyme activity impairs performance. The concept of sports anaemia can be criticised. Simply measuring the blood levels does not take into account the haemodilution that occurs in athletes because of training. The lack of these measurements makes it difficult to diagnose anaemia or evaluate any treatment. Anaemia is treated by preventing decreased iron stores through a balanced food intake or iron supplements. Self-medications must be discouraged because of intolerance, risk of overdose and many other drug interactions. PMID- 10367336 TI - Stimulating research to improve the scientific basis of risk assessment. PMID- 10367334 TI - Reliability and validity of measures of cardiac output during incremental to maximal aerobic exercise. Part II: Novel techniques and new advances. AB - For exercise physiologists and sport cardiologists, one of the greatest challenges is to develop a valid, reliable, noninvasive and affordable measure of cardiac output (Q). There are several techniques available to measure Q during exercise conditions. These procedures generally provide accurate and reliable determinations of Q during submaximal exercise, but may be limited during maximal exercise conditions. The most commonly used noninvasive measures are the acetylene (C2H2) and carbon dioxide (CO2) rebreathe methods as reviewed in part I of this article. Only the foreign gas rebreathe method, using C2H2, meets all of the criteria of being noninvasive, easy to use, reliable and valid for use during maximal exercise. New methodologies have recently been developed to measure Q during exercise conditions. Although not as popular as the C2H2 and CO2 rebreathe methods, these methods have increasingly gained favour in exercise physiology and sport cardiology settings. The majority of these measures (if performed meticulously), with the exception of impedance cardiography, provide reasonably accurate and reliable determinations of Q. However, the cost of usage and technological limitations during maximal exercise have prevented these techniques from replacing the conventional measures of Q during exercise conditions. Doppler echocardiography and the modified C2H2 methods hold promise for the assessment of Q during maximal exercise. With further advances in these technologies their use in exercise physiology and sport cardiology setting may become more common. PMID- 10367337 TI - The enigma of arsenic carcinogenesis: role of metabolism. AB - Inorganic arsenic is considered a high-priority hazard, particularly because of its potential to be a human carcinogen. In exposed human populations, arsenic is associated with tumors of the lung, skin, bladder, and liver. While it is known to be a human carcinogen, carcinogenesis in laboratory animals by this metalloid has never been convincingly demonstrated. Therefore, no animal models exist for studying molecular mechanisms of arsenic carcinogenesis. The apparent human sensitivity, combined with our incomplete understanding about mechanisms of carcinogenic action, create important public health concerns and challenges in risk assessment, which could be met by understanding the role of metabolism in arsenic toxicity and carcinogenesis. This symposium summary covers three critical major areas involving arsenic metabolism: its biodiversity, the role of arsenic metabolism in molecular mechanisms of carcinogenesis, and the impact of arsenic metabolism on human risk assessment. In mammals, arsenic is metabolized to mono- and dimethylated species by methyltransferase enzymes in reactions that require S adenosyl-methionine (SAM) as the methyl donating cofactor. A remarkable species diversity in arsenic methyltransferase activity may account for the wide variability in sensitivity of humans and animals to arsenic toxicity. Arsenic interferes with DNA methyltransferases, resulting in inactivation of tumor suppressor genes through DNA hypermethylation. Other studies suggest that arsenic induced malignant transformation is linked to DNA hypomethylation subsequent to depletion of SAM, which results in aberrant gene activation, including oncogenes. Urinary profiles of arsenic metabolites may be a valuable tool for assessing human susceptibility to arsenic carcinogenesis. While controversial, the idea that unique arsenic metabolic properties may explain the apparent non-linear threshold response for arsenic carcinogenesis in humans. In order to address these outstanding issues, further efforts are required to identify an appropriate animal model to elucidate carcinogenic mechanisms of action, and to define dose response relationships. PMID- 10367335 TI - Nerve entrapment syndromes as a cause of pain in the hip, groin and buttock. AB - In sports medicine, chronic hip, groin and buttock pain is a common diagnostic problem. Because of the complex anatomy of this region and the many potential neurological causes for pain, few sports clinicians have a detailed understanding of this problem. This paper discusses the clinical aspects of nerve entrapment syndromes related to sport and takes a regional approach in order to provide a diagnostic framework for the general sports physician. The various neurological syndromes are discussed and the surgical management elaborated in detail. For some specific conditions, such as the so-called 'piriformis syndrome', the pathophysiological understanding has changed since the early descriptions and now this particular diagnosis is often ascribed to almost any cause of buttock and/or hamstring symptoms. We discuss the nature of the origin of local symptoms and note that the often described symptoms are more likely due to compression of structures other then the sciatic nerve. Furthermore, the role of piriformis hypertrophy or anatomical nerve variations in the genesis of this syndrome must be questioned. We suggest renaming this the 'deep gluteal syndrome' to account for all of the observed phenomena. As sports medicine continues to develop a scientific basis, the role of nerve entrapments as the basis for chronic symptomatology is undergoing a new understanding and clinicians need to be aware of the diagnostic possibilities and be able to advise patients accordingly on the basis of scientific fact not anecdotal fiction. PMID- 10367339 TI - Distribution of uranium in rats implanted with depleted uranium pellets. AB - During the Persian Gulf War, soldiers were injured with depleted uranium (DU) fragments. To assess the potential health risks associated with chronic exposure to DU, Sprague Dawley rats were surgically implanted with DU pellets at 3 dose levels (low, medium and high). Biologically inert tantalum (Ta) pellets were used as controls. At 1 day and 6, 12, and 18 months after implantation, the rats were euthanized and tissue samples collected. Using kinetic phosphorimetry, uranium levels were measured. As early as 1 day after pellet implantation and at all subsequent sample times, the greatest concentrations of uranium were in the kidney and tibia. At all time points, uranium concentrations in kidney and bone (tibia and skull) were significantly greater in the high-dose rats than in the Ta control group. By 18 months post-implantation, the uranium concentration in kidney and bone of low-dose animals was significantly different from that in the Ta controls. Significant concentrations of uranium were excreted in the urine throughout the 18 months of the study (224 +/- 32 ng U/ml urine in low-dose rats and 1010 +/- 87 ng U/ml urine in high-dose rats at 12 months). Many other tissues (muscle, spleen, liver, heart, lung, brain, lymph nodes, and testicles) contained significant concentrations of uranium in the implanted animals. From these results, we conclude that kidney and bone are the primary reservoirs for uranium redistributed from intramuscularly embedded fragments. The accumulations in brain, lymph nodes, and testicles suggest the potential for unanticipated physiological consequences of exposure to uranium through this route. PMID- 10367338 TI - A physiological model for tert-amyl methyl ether and tert-amyl alcohol: hypothesis testing of model structures. AB - The oxygenate tert-amyl methyl ether (TAME) is a gasoline fuel additive used to reduce carbon monoxide in automobile emissions. To evaluate the relative health risk of TAME as a gasoline additive, information is needed on its pharmacokinetics and toxicity. The objective of this study was to use a physiologically-based pharmacokinetic (PBPK) model to describe the disposition of TAME and its major metabolite, tert-amyl alcohol (TAA), in male Fischer-344 rats. The model compartments for TAME and TAA were flow-limited. The TAME physiological model had 6 compartments: lung, liver, rapidly perfused tissues, slowly perfused tissues, fat, and kidney. The TAA model had 3 compartments: lung, liver, and total-body water. The 2 models were linked through metabolism of TAME to TAA in the liver. Model simulations were compared with data on blood concentrations of TAME and TAA taken from male Fischer-344 rats during and after a 6-hour inhalation exposure to 2500, 500, or 100 ppm TAME. The PBPK model predicted TAME pharmacokinetics when 2 saturable pathways for TAME oxidation were included. The TAA model, which included pathways for oxidation and glucuronide conjugation of TAA, underpredicted the experimental data collected at later times postexposure. To account for biological processes occurring during this time, three hypotheses were developed: nonspecific binding of TAA, diffusion-limited transport of TAA, and enterohepatic circulation of TAA glucuronide. These hypotheses were tested using three different model structures. Visual inspection and statistical evaluation involving maximum likelihood techniques indicated that the model incorporating nonspecific binding of TAA provided the best fit to the data. A correct model structure, based upon experimental data, statistical analyses, and biological interpretation, will allow a more accurate extrapolation to humans and, consequently, a greater understanding of human risk from exposure to TAME. PMID- 10367340 TI - Metabolism of [14C]phenol in the isolated perfused mouse liver. AB - A previous report from this laboratory focused on the metabolism of [14C]benzene (BZ) in the isolated, perfused, mouse liver (C. C. Hedli, et al., 1997, Toxicol. Appl. Pharmacol. 146, 60-68). Whereas administration of BZ to mice results in bone marrow depression (R. Snyder et al., 1993, Res. Commun. Chem. Pathol. Pharmacol. 20, 191-194), administration of phenol (P), the major metabolite of BZ, does not. It was, therefore, of interest to determine whether the metabolic fate of P produced during BZ metabolism differed from that of P metabolized in the absence of BZ. Mouse livers were perfused with a solution of [14C]P in both the orthograde (portal vein to central vein) and retrograde (central vein to portal vein) direction to investigate the metabolic zonation of enzymes involved in P hydroxylation and conjugation. Perfusate samples were collected, separated by HPLC, and tested for radioactivity. Unconjugated metabolites were identified by comparing their retention times with nonradiolabeled standards, which were detected by UV absorption. Conjugated metabolites were identified and collected on the basis of radiochromatogram results, hydrolyzed enzymatically, and identified by co-chromatography with unlabeled BZ metabolites. The objective was to compare and quantify the metabolites formed during the perfusion of P in the orthograde and retrograde directions and to compare the orthograde P-perfusion results with the orthograde BZ results reported previously. Regardless of the direction of P perfusion, the major compounds released from the liver were P. phenylgucuronide, phenylsulfate, hydroquinone (HQ), and HQ glucuronide. A comparison of the results of perfusing P in the orthograde versus the retrograde direction showed that more P was recovered unchanged and more HQ was formed during retrograde perfusion. The results suggest that enzymes involved in P hydroxylation are generally closer to the central vein than those involved in conjugation, and that during retrograde perfusion, P metabolism may be limited by the sub-optimal conditions of perfusion. Comparison of the orthograde perfusion studies of P and BZ revealed that a larger percentage of the radioactivity released from the liver was identified as unconjugated HQ after BZ perfusion than after P perfusion. In addition, the amount of radioactivity covalently bound to liver macromolecules was measured after each perfusion and determined to be proportional to the amount of HQ and HQG detected in the perfusate samples. PMID- 10367341 TI - The role of dermal irritation in the skin tumor promoting activity of petroleum middle distillates. AB - Petroleum middle distillates (PMDs), a class of hydrocarbons which boil between 350-700 degrees F, are tumor promoters in mouse skin. The promotional activity is produced under conditions that also result in local changes, including chronic irritation and epidermal hyperplasia. The present study was conducted by comparing equal weekly doses of irritating and minimally or nonirritating test materials, to assess whether tumor promotion was a secondary response to these effects. Four PMDs, C10-C14 normal paraffins (NP), lightly refined paraffinic oil (LRPO), Jet Fuel A (JF), and steam-cracked gas oil (SCGO), were evaluated. Test materials were applied undiluted (2x/week) or as 28.6% (7x/week) or 50% (4x/week) concentrations in mineral oil for 52 weeks following initiation with dimethylbenzanthracene (DMBA). When applied undiluted, all materials produced moderate irritation and significant increase in tumor incidence. When NP, LRPO, or JF were applied in mineral oil diluent, skin irritation was generally ameliorated and few, if any, tumors were produced. SCGO was irritating and produced a significant increase in tumor frequency when administered in mineral oil diluent. These data indicate that the promotional activity of straight-run PMDs is likely related to chronic irritation at the application site and not to dose. Thus, when used appropriately in the absence of prolonged irritation, these materials should not present a tumorigenic hazard to humans. PMID- 10367342 TI - Inhalation toxicity and carcinogenicity studies of cobalt sulfate. AB - Cobalt sulfate is a water-soluble cobalt salt with a variety of industrial and agricultural uses. Several cobalt compounds have induced sarcomas at injection sites in animals, and reports have suggested that exposure to cobalt-containing materials may cause lung cancer in humans. The present studies were done because no adequate rodent carcinogenicity studies had been performed with a soluble cobalt salt using a route relevant to occupational exposures. Groups of 50 male and 50 female F344/N rats and B6C3F1 mice were exposed to aerosols containing 0, 0.3, 1.0, or 3.0 mg/m3 cobalt sulfate hexahydrate, 6 h/day, 5 days/week, for 104 weeks. Survival and body weights of exposed rats and mice were generally unaffected by the exposures. In rats, proteinosis, alveolar epithelial metaplasia, granulomatous alveolar inflammation, and interstitial fibrosis were observed in the lung in all exposed groups. Nonneoplastic lesions of the nose and larynx were also attributed to exposure to all concentrations of cobalt sulfate. In 3.0 mg/m3 male rats and in female rats exposed to 1.0 or 3.0 mg/m3, the incidences of alveolar/bronchiolar neoplasms were increased over those in the control groups. Lung tumors occurred with significant positive trends in both sexes. The incidences of adrenal pheochromocytoma in 1.0 mg/m3 male rats and in 3.0 mg/m3 female rats were increased. Nonneoplastic lesions of the respiratory tract were less severe in mice than in rats. In mice, alveolar/bronchiolar neoplasms in 3.0 mg/m3 males and females were greater than those in the controls, and lung tumors occurred with significantly positive trends. Male mice had liver lesions consistent with a Helicobacter hepaticus infection. Incidences of liver hemangiosarcomas were increased in exposed groups of male mice; however, because of the infection, no conclusion could be reached concerning an association between liver hemangiosarcomas and cobalt sulfate. In summary, exposure to cobalt sulfate by inhalation resulted in increased incidence of alveolar/bronchiolar neoplasms and a spectrum of inflammatory, fibrotic, and proliferative lesions in the respiratory tracts of male and female rats and mice. Adrenal pheochromocytomas were increased in female rats, and possibly in male rats. PMID- 10367344 TI - Organophosphorus-induced neurotoxicity in the absence of neuropathy target esterase inhibition: the effects of triphenyl phosphine in the European ferret. AB - Abou-Donia et al. (in Toxicologist, Vol. 30, 1996) have reported that repeated oral administration of the organo-phosphorus compound triphenyl phosphine (TPPn) to the domestic chicken results in neuropathological changes in the spinal cord and peripheral nerves, accompanied by ataxia and paralysis. This study also noted that single doses of TPPn resulted in no inhibition of the enzymes neuropathy target esterase (NTE) and acetylcholinesterase (AChE). We undertook the present study to determine the biochemical, neuropathological, and clinical effects of single doses of TPPn in the European ferret, a mammalian species shown to be susceptible to organophosphorus-induced neurotoxicity. Eight 12-week-old ferrets were each injected subcutaneously with either 250 mg TPPn/kg bw or 500 mg TPPn/kg bw, or with the peanut oil/ethyl ether vehicle. Twenty-four h after dosing, the brains of 5 animals from each dose group were examined for NTE and AChE activities. The remaining 3 animals in each group were observed for 6 days for the development of clinical signs, after which their brains were processed for the presence of axonal degeneration using the Fink-Heimer silver impregnation method. Single injections of TPPn had no effect on the activities of whole-brain NTE or AChE 24 h after injection. The animals observed for clinical signs showed increasing trunk and hindlimb ataxia beginning 4 days after injection, culminating in fore-and hindlimb paralysis 6 days after injection. All brains exposed to either dose of TPPn showed widespread axonal degeneration extending from the brainstem and cerebellum into midbrain and forebrain areas. The results of this study support the hypothesis that TPPn-induced neurotoxicity is a separate and distinct form of organophosphorus-induced neurotoxicity not dependent on NTE inhibition, and therefore not a variant of organophosphorus induced delayed neurotoxicity (OPIDN). PMID- 10367343 TI - Activated human T lymphocytes exhibit reduced susceptibility to methylmercury chloride-induced apoptosis. AB - Mercurials have been shown to cause apoptosis in human T cells. The objective of this study was to evaluate and compare the relative susceptibility of resting versus activated T cells to methyl mercury chloride (MeHgCl)-induced cell death. Apoptosis was assessed by Hoechst 33258 and 7-AAD staining and annexin V binding. Our results show that activation of T cells by PHA, PMA, and ionomycin, or IL-2, reduces mercury-induced apoptosis by approximately 50%. We have previously shown that the underlying basis for these toxic effects involves perturbation of mitochondrial function leading to oxidative stress and the release of cytochrome c to the cytosol. Therefore, the ability of MeHgCl to alter the mitochondrial transmembrane potential (delta psi m) and to induce the generation of reactive oxygen species (ROS) was evaluated in activated T-cells. Both resting and activated cells treated with MeHgCl exhibited a decrease in delta psi m when compared to respective control cells. ROS production was elevated in resting cells following treatment with mercury; in contrast, fewer activated T cells exhibit increased levels of ROS in the presence of MeHgCl. Similarly, MeHgCl treatment resulted in the release of cytochrome c to the cytoplasm in non activated T cells but failed to do so in the activated population. These results lead us to examine intracellular levels of bcl-2, a protein that has been shown to regulate apoptosis, presumably via its ability to associate with the mitochondrial membrane. Bcl-2 levels were found, in resting cells, to be low in the presence or absence of mercury. In comparison, activated T cells expressed elevated levels of bcl-2. The relationship between mercury-induced apoptosis in human T cells, mitochondrial dysfunction, and intracellular levels of bcl-2 are discussed. PMID- 10367345 TI - Factors contributing to the acute and subchronic adverse respiratory effects of machining fluid aerosols in guinea pigs. AB - Several physical, chemical, and microbial factors are potential contributors to the adverse pulmonary effects associated with occupational exposure to machining fluid aerosols. The present study examined the relative toxicity of 3 major classes of machining fluids (soluble, semi-synthetic, and synthetic) as well as that of unused (fresh) versus used (grab samples taken from manufacturing sites) machining fluids. Pulmonary function and changes in cellular and biochemical indices in bronchoalveolar lavage fluid were examined during and 24 h after exposure, respectively. Statistically significant differences in toxicity were observed in guinea pigs exposed for 3 h to respirable aerosols of unused machining fluids (semi-synthetic > soluble >> synthetic). In addition, greater toxicity was observed in animals exposed to used, machining fluid aerosols compared to unused fluids. Moreover, within the used machining fluid types, significantly greater adverse effects were observed in animals exposed to poorly maintained fluids (i.e., heavy microbial contamination) versus well-maintained fluids. Changes in biochemical and cellular parameters in bronchoalveolar lavage fluid occurred after a single exposure to 5 mg/m3 of the poorly maintained used machining fluid aerosols. Changes in inflammation but not LDH and protein were observed in animals repeatedly exposed to semi-synthetic machining fluid aerosols. A statistically significant increase in lavage fluid neutrophils was observed in guinea pigs exposed to 5 mg/m3 used, semi-synthetic machining fluid aerosols for 4 weeks. In separate experiments, physicochemical properties of unused machining fluids were found to contribute to the production of adverse effects. Adjustment of the alkaline and hypotonic nature of the unused semi synthetic machining fluid to isotonicity and pH 7 significantly reduced adverse effects. Together, these findings strongly suggest that multiple factors contribute to the adverse respiratory effects associated with occupational exposure to machining fluid aerosols. PMID- 10367347 TI - Delayed acute toxicity of 1,2,3,4,6,7,8-heptachlorodibenzo-p-dioxin (HpCDD), after oral administration, Obeys Haber's rule of inhalation toxicology. AB - Eight different doses (2.5 to 10.0 mg/kg) of 1,2,3,4,6,7,8-heptachlorodibenzo-p dioxin (HpCDD) were administered acutely to a total of 272 female Sprague-Dawley rats. The doses ranged from a NOAEL for wasting/hemorrhage to supralethal doses. Dose- and time-responses of wasting/hemorrhage, anemia, and cancer were and are being studied as end points of toxicity. The experiments will be continued until the last rat dies. There was a very steep dose- and time-response between the LOAEL for wasting/hemorrhage (2.8 mg/kg) and the third highest dose (4.1 mg/kg) of HpCDD. The dose-and time-responses were nearly symmetrical, obeying Haber's Rule of inhalation toxicology (c x t = constant) even beyond 100% mortality. Introduction of a minimum of 25% body weight loss as a discriminatory criterion to separate wasting from hemorrhage as the primary cause of death reduced variability from 5.8 to 3.2%. An arithmetic plot of the dose and time data resulted in a nearly perfect hyperbola. A logarithmic plot of these data yielded a straight line of similar perfection. Dose-response data at constant times illustrate the shifting of the dose-response curve towards a liminal value, which represents the necessary observation period for this effect. Time-response data at constant doses demonstrate the shifting of the time-response curve towards a liminal value, which represents the LOAEL for the dose-response of this effect. A three-dimensional plot of dose- and time-response data depicts the surface area on which c x t is constant along hyperbolas, in terms of wasting as the end point of toxicity. Surviving rats in all groups started developing anemia 126 days after dosing, but no rat died of wasting/hemorrhage after day 74. Rats surviving anemia began to die of lung cancer as of day 397 after dosing. Thus, although the experiment has been completed as far as dose- and time-responses of wasting/hemorrhage are concerned, it will be about another 2 years before complete dose and time responses will become available for anemia and lung cancer. PMID- 10367346 TI - Comparison of pulmonary and pleural responses of rats and hamsters to inhaled refractory ceramic fibers. AB - The present study was designed to determine whether pleural fiber burdens or subchronic pleural fibroproliferative and inflammatory changes can help explain the marked interspecies differences in pleural fibrosis and mesothelioma that are observed following long-term inhalation of RCF-1 ceramic fibers by rats and hamsters. Fischer 344 rats and Syrian golden hamsters were exposed to RCF-1 for 4 h per day, 5 days per week, for 12 consecutive weeks. Lung and pleural fiber burdens were characterized during and after exposure. For all time points, approximately 67% of fibers associated with lung tissues from both rats and hamsters were longer than 5 microns in length. In comparison, fibers longer than 5 microns recovered from the pleural compartment, following a 12-week exposure and 12 weeks of recovery, accounted for 13% (hamsters) and 4% (rats) of the distribution. In the 12 weeks after the cessation of exposure, the number of fibers longer than 5 microns in length remained constant in the hamster at approximately 150 fibers per cm2 pleura. This was 2 to 3 times the corresponding fiber surface density in the rat. Significant pulmonary and pleural inflammation was detected at all time points and for both species. DNA synthesis by pleural mesothelial cells was quantified by bromodeoxyuridine uptake following 3 days of labeling. Labeling indices were higher in hamsters than in rats, both for RCF-1 exposed and filtered air-control animals and was highest for the parietal surface of the pleura. Significantly greater collagen deposition was measured in the visceral pleura of hamsters 12 weeks post-exposure but was not significantly elevated in rats. These findings demonstrate that subchronic inhalation exposure to RCF-1 induces pleural inflammation, mesothelial-cell turnover, pleural fibrosis, and an accumulation of fibers with a length greater than 5 microns in the hamster. The accumulation of long fibers in the pleural space may contribute to the pathology observed in the hamster following chronic inhalation of RCF-1, whereas the presence of short, thin fibers may play a role in the acute-phase biological response seen in both species. PMID- 10367348 TI - Threshold dose response for tumor induction by genotoxic carcinogens modeled via cell-cycle delay. AB - Dose-response relationships for tumor induction in animal bioassays for carcinogenicity are often postulated to include thresholds, particularly for nongenotoxic chemicals that increase the rate of cell proliferation at high doses. In this report, thresholds are postulated also for genotoxic carcinogens. The hypothesis is based on the idea of a delay of the cell cycle induced by low level DNA damage and an acceleration at cytotoxic dose levels, thus resulting in a J-shaped (or U-shaped) dose response for cell turnover. Calculations were based on the 2-stage clonal expansion model of carcinogenesis. The background values chosen for the model parameters resulted in a 10.5% "spontaneous" 2-year cumulative tumor incidence. Using this as a starting point, a decrease by 3, 10, and 30% in the rates of cell turnover resulted in a decrease in the spontaneous tumor incidence to 9.4, 7.1 and 3.0%, respectively. Dose-responses with J-shaped curves for the rates of cell birth and death were modeled by shifted quadratic functions reaching the minimum at dose 1. Combinations with linearly increasing mutation rates also generated, under certain conditions, J-shaped dose-response curves for tumor incidence. As an example, for a 30% increase in mutation rates and a 10% decrease in cell turnover rates (both at dose 1), the dose-response curve showed an initial decrease of tumor incidence below the spontaneous rate, a reversion to the background value at 0.8 dose units, and an increase thereafter. The 0.8 dose could be considered to represent the "threshold dose." The approach presented might reconcile opposing views on thresholds on a biologically plausible mechanistic basis, and show a way for the quantitative estimation of threshold doses. PMID- 10367349 TI - Localization and comparative toxicity of methylsulfonyl-2,5- and 2,6 dichlorobenzene in the olfactory mucosa of mice. AB - Several methylsulfonyl (MeSO2) metabolites formed from chlorinated aromatic hydrocarbons have been identified in human milk, lung, and body fat, as well as in the tissues of Baltic grey seals and arctic polar bears. The tissue localization and nasal toxicity of two methylsulfonyl-substituted dichlorobenzenes (diCl-MeSO2-B), with the chlorine atoms in the 2,5-, and 2,6- positions, were investigated in female NMRI and C57B1 mice. Using tape-section autoradiography, animals dosed i.v. with 14C-labeled 2,5-, or 2,6-(diCl-MeSO2-B) showed a preferential uptake of radioactivity in the olfactory mucosa and the tracheobronchial epithelium. Histopathology showed that 2,6-(diCl-MeSO2-B) is a potent toxicant that induces necrosis in the olfactory mucosa following a single dose as low as 4 mg/kg (i.p. injection), whereas 2,5-(diCl-MeSO2-B) induced no signs of toxicity in the olfactory mucosa at doses as high as 130 mg/kg (i.p. injection). Necrosis of the Bowman's glands was the first sign of 2,6-(diCl-MeSO2 B)-induced toxicity followed by degeneration of the neuroepithelium, which implies that the Bowman's gland may be the primary site of toxicity and degeneration of the neuroepithelium may be a secondary effect. Administration of the parent compounds, 1,3-dichlorobenzene and 1,4-dichlorobenzene, or the chlorinated analog 1,2,3-trichlorobenzene (85, 85, and 105 mg/kg, respectively; i.p. injection), induced no signs of toxicity in the olfactory mucosa. These and previous results suggest that 2,6-positioned chlorine atoms and an electron withdrawing substituent in the primary position is an arrangement that predisposes for toxicity in the olfactory mucosa. PMID- 10367350 TI - Exposure of C57BL/6 mice to carbon disulfide induces early lesions of atherosclerosis and enhances arterial fatty deposits induced by a high fat diet. AB - Even though atherosclerotic cardiovascular disease (ACVD) is the number one cause of death in the United States, the effects of environmental toxicants on this process are less well studied than the effects of chemicals on the second leading cause of death, cancer. There is considerable epidemiological evidence that workers exposed to carbon disulfide (CS2) have increased rates of ACVD, and there is conflicting evidence of the atherogenic potential of CS2 from animal studies. Chemical modification, such as oxidation of low-density lipoproteins (LDL), is tightly associated with increased LDL uptake by macrophages and the development of arterial fatty streaks. CS2 has been previously demonstrated to modify several proteins in vitro including LDL, and others in vivo through derivatization and covalent cross-linking. To investigate both the capacity of CS2 to induce arterial fatty deposits by itself, and its ability to enhance the rate of fatty deposit formation induced by a western style, high fat diet, groups of 20 female C57BL/6 mice were exposed to 0, 50, 500, or 800 ppm CS2 by inhalation. Half the animals in each group were placed on an atherogenic high fat diet and half on a control diet (NIH-07). Animals were sacrificed after 1, 4, 8, 12, 16, or 20 weeks of exposure, and the rates of fatty deposit formation under the aortic valve leaflets were evaluated. Exposure of mice on the control diet to 500 and 800 ppm CS2 induced a small but significant increase in the rate of fatty deposit formation over non-exposed controls. A more striking result was observed in the animals on the high fat diet. There was marked enhancement of the rate of fatty deposit formation in mice exposed to 500 and 800 ppm over the animals on the high fat diet alone. In addition, there was a small but significant enhancement in mice exposed to 50 ppm over the rate of fatty deposit formation induced by the high fat diet alone. Analysis of erythrocyte spectrin for protein cross-linking revealed a dose-dependent formation of alpha- and beta-heterodimers in animals on both diets. These data demonstrate that CS2 is atherogenic at high concentrations, but more importantly, suggest that, in conjunction with other risk factors, CS2 at relatively low concentrations can enhance atherogenesis. PMID- 10367351 TI - The toxic and metabolic effects of 23 aliphatic alcohols in the isolated perfused rat liver. AB - We investigated the acute toxic and metabolic effects of 23-aliphatic alcohols (16 saturated and 7 unsaturated) in the isolated perfused rat liver at a concentration of 65.1 mmol/l (approximately 0.3% ethanol). The capacity of the straight chain primary alcohols (methanol, ethanol, 1-propanol, 1-butanol and 1 pentanol) to release the enzymes glutamate-pyruvate transaminase (GPT), lactate dehydrogenase (LDH) and glutamate dehydrogenase (GLDH) into the perfusate was strongly correlated with their carbon chain length. The secondary alcohols were less active in this respect whereas branching of the carbon chain did not consistently change alcohol toxicity. Unsaturation in the straight chain but not in the branched chain alcohols was accompanied by an increase in toxicity. An increased enzyme release was in general accompanied by, and correlated to, reductions in oxygen consumption, bile secretion, and perfusion flow of the isolated livers. Statistically significant correlations exist between parameters of alcohol-induced hepatotoxicity and the membrane/buffer partition coefficents of the alcohols. With the exception of methanol, all alcohols tested increased the lactate/pyruvate ratio of the perfusate, although this effect was not correlated to the degree of hepatic injury. Hepatic ATP concentrations decreased in most cases in line with hepatic injury and were particularly correlated with changes in oxygen consumption. Hepatic concentrations of reduced glutathione (GSH) were only diminished by the unsaturated alcohols, whereas an increase in hepatic oxidized glutathione (GSSG) occurred only with some of the saturated alcohols. Hepatic concentrations of malondialdehyde (MDA) increased after two saturated and three unsaturated alcohols but did not correlate with other parameters of hepatotoxicity. In conclusion, alcohol-induced hepatotoxicity is primarily due to membrane damage induced by the direct solvent properties of the alcohols. The consequences and relative contributions of alcohol metabolization to the overall hepatotoxicity of higher alcohols requires further study. PMID- 10367352 TI - Virulence gene regulation in pathogenic Escherichia coli. AB - The ability to regulate gene expression throughout the course of an infection is important for the survival of a pathogen in the host. Thus, virulence gene expression responds to environmental signals in many complex ways. Frequently, global regulatory factors associated with specific regulators co-ordinate expression of virulence genes. In this review, we present well-described regulatory mechanisms used to co-ordinate the expression of virulence factors by pathogenic Escherichia coli with a relative emphasis on diseases caused by E. coli in animals. Many of the virulence-associated genes of pathogenic E. coli respond to environmental conditions. The involvement of global regulators, including housekeeping regulons and virulence regulons, specific regulators and then sensor regulatory systems involved in virulence, is described. Specific regulation mechanisms are illustrated using the regulation of genes encoding for fimbriae, curli, haemolysin and capsules as examples. PMID- 10367353 TI - Pathogenic diversity of Escherichia coli and the emergence of 'exotic' islands in the gene stream. AB - Escherichia coli is a highly adaptive bacterial species that is both a member of the commensal intestinal flora and a versatile pathogen associated with numerous types of intestinal and systemic infections in humans and other animals. The spectrum of diseases caused by E. coli is due to the acquisition of specific virulence genes harbored on plasmids, bacteriophages, or within distinct DNA segments termed pathogenicity islands (PAIs) that are absent from the genomes of commensal E. coli strains. PAIs are likely to have been transferred horizontally and may have integrated into the E. coli chromosome through bacteriophage or plasmid integration or transposition. The contribution of intergenic inheritance to the adaptation and evolution of E. coli, types of PAIs associated with different groups of pathogenic E. coli and approaches to identify unique sequence islands (USIs), some of which might confer pathogenicity, in E. coli and other bacteria are presented. PMID- 10367354 TI - Secretion of virulence factors by Escherichia coli. AB - In order to interact with their host, pathogenic strains of E. coli need to secrete some virulence factors which can modify the metabolism of host cells, contributing to disease. Since E. coli is a Gram-negative bacteria, this secretion process involves the crossing of both the inner and the outer membranes. E. coli uses mainly four secretion mechanisms called type I, type II, type III and type IV secretion systems. In the type I secretion system, the secretion machinery is composed of three proteins forming a channel through the inner and outer membranes. It is a one-step mechanism. The secretion signal is present in the carboxyterminal region of the secreted protein but without proteolytic cleavage. In E. coli, the best studied type I secreted protein is haemolysin. In type II and type IV secretion systems, the crossing of the inner membrane involves the sec machinery with the cleavage of an aminoterminal signal sequence. The crossing of the outer membrane involves the formation of a pore either by other proteins (type II) or by the carboxyterminal region of the protein (type IV). The A-B toxins, such as heat labile enterotoxin, are secreted by the first mechanism and members of the IgA proteases are secreted by the second. The type III secretion system involves at least 20 proteins including cytoplasmic, inner membrane and outer membrane proteins. The originality of this system is the ability to inject secreted bacteria into the cytosol of the host cells. Such a system is found in attaching and effacing E. coli and in diffusely adhering E. coli. PMID- 10367355 TI - Rabbit EPEC: a model for the study of enteropathogenic Escherichia coli. AB - Colibacillosis has become, in rational rabbit breeding units of western Europe, one of the most economically and pathologically important issues since the beginning of the 1980s. Data on the virulence mechanisms and the phenotypic characters of the E. coli strains that are responsible for lethal diarrhoea epizootics have been gathered throughout the years. These strains are representative of a pathovar called enteropathogenic E. coli (EPEC) in diarrhoeagenic strains of human origin. EPEC are mainly characterized by their ability to induce a typical lesion called attachment/effacement, whose determinism lies in a pathogenicity island: the locus of enterocyte effacement. The understanding of the pathogenesis mechanisms of this type of bacteria should lead to new tools helping to control the disease in rabbit farming. PMID- 10367356 TI - Necrotoxic Escherichia coli (NTEC): two emerging categories of human and animal pathogens. AB - Necrotoxic Escherichia coli (NTEC) were originally defined as strains of E. coli producing a toxin called cytotoxic necrotising factor (CNF). Two types of CNF have been identified, each of them being genetically linked to several other specific virulence markers, a situation that allows the definition of two distinct homogeneous categories of NTEC called NTEC-1 and NTEC-2. CNF1 and CNF2 are highly homologous holoproteins containing 1,014 amino acids that exert both lethal and necrotic activities in vivo and induce multinucleation and actin stress fibres in cell cultures. The activity of CNFs on mammal cells is due to their ability to constitutively activate by deamidation the Rho proteins, a family of small GTPases that regulate the physiology of the cell cytoskeleton. In NTEC-1, the gene encoding CNF1 belongs to a pathogenicity island which also comprises the genes encoding for alpha-haemolysin and P-fimbriae. In NTEC-2 strains, CNF2 is encoded by a plasmid that also encodes, in 100% of the isolates, a new member of the cytolethal distending toxin family (CDT-III) and in about 50% of the isolates, the F17b-fimbrial adhesin that confers the ability to adhere to calf intestinal villi. The presence of CDT is also suspected in a large majority of NTEC-1 strains. NTEC-1 strains can be found in humans and in all species of domestic mammals, whereas NTEC-2 strains have only been reported in ruminants. The implication of NTEC strains has been clearly established in extra-intestinal infections of humans and animals, for instance in urinary tract infections for NTEC-1 strains. Their role in severe dysenteric syndromes, both in humans and animals, is substantiated by several clinical reports, but there is little published information on this pathogenicity in animal models of infection. The combined production of several powerful toxins (haemolysin, CNF, CDT) by NTEC strains makes them, however, potentially aggressive pathogens which deserve to be searched for on a larger scale. Moreover, NTEC-1 from man and animals appear to be highly related according to available molecular markers, which indicates that domestic animals could constitute reservoirs of NTEC strains which are pathogenic for humans. PMID- 10367357 TI - Shiga/verocytotoxins and Shiga/verotoxigenic Escherichia coli in animals. AB - Vero/Shiga toxins (VT/Stx) have an A-B structure: the A subunit carries the enzymatic activity and the B subunit binds the toxin to the membrane receptor (Gb3 or Gb4). The VT/Stx inhibit protein synthesis in the target eucaryotic cells, mainly the endothelial cells of blood vessels. The VT/Stx are subdivided into two families. VT1/Stx1 is a homogeneous family of toxins identical to the Stx of Shigella dysenteriae. VT2/Stx2 is a more heterogeneous family of toxins more distantly related to this Stx toxin. The VT2/Stx2 variants can be distinguished by the polymerase chain reaction (PCR) and/or the reaction with monoclonal antibodies. The VT/Stx-producing Escherichia coli are also subdivided into two main groups on the basis of the presence or absence of additional properties: the enterohaemorrhagic E. coli (EHEC) induce the formation of attaching/effacing lesions and carry a 60 MD plasmid encoding a specific haemolysin (the enterohaemolysin); the vero/shiga-toxigenic E. coli (VTEC/STEC) do not show these properties. The EHEC are isolated from humans and ruminants, especially young calves. They are associated with haemorrhagic enterocolitis and its sequelae in humans, the haemolytic-uraemic syndrome (HUS). The VT/Stx play a role in the occurrence of blood in the faeces and in the HUS by their action on the endothelial cells of blood vessels in the intestinal submucosa and in the renal glomeruli, after resorption through the intestinal walls. The VTEC/STEC are isolated from piglets, calves and humans. In recently weaned piglets, they cause the oedema disease, an enterotoxaemia characterized by subcutaneous, mesenteric and cerebral oedemas, with nervous disorders as main clinical signs. The oedema disease is the consequence of the action of the VT/Stx on the endothelial cells of blood vessels in various organs. In calves and humans, the role in disease of VTEC/STEC is controversial, but they could be associated with some cases of diarrhoea and HUS. The case of the O157:H7 EHEC which are present in healthy cattle of various ages, but are highly virulent for humans is of special interest. The potential zoonotic aspect of VT/Stx-producing E. coli infections in animals is detailed chapter by chapter. Prophylaxis of these infections by vaccination is the subject of the discussion on the future of the research studies on these pathogenic bacteria. PMID- 10367358 TI - Enterotoxigenic Escherichia coli (ETEC) in farm animals. AB - Animal diseases due to enterotoxigenic Escherichia coli (ETEC) typically appear as severe watery diarrhoea during the first few days of life (also a few days after weaning in pigs). ETEC adhere to the small intestinal microvilli without inducing morphological lesions and produce enterotoxins acting locally on enterocytes. This action results in the hypersecretion (of water and electrolytes) and reduced absorption. Adhesins and toxins are the two prominent virulence attributes of ETEC and the level of knowledge of these factors determines the chances for successful prevention and therapy of the disease. For animal ETEC the most common adhesins are the fimbriae (pili) on the surface: F4(K88), F5(K99), F6(987P), F41, F42, F165, F17 and F18. Enterotoxins (extracellular proteins or peptides) of animal ETEC are classified as heat-labile (LT) and heat-stable (ST) enterotoxins with further subdivisions (LTh-I, LTp-I, LTIIa, LTIIb, STaH, STaP, STb) according to antigenic and biological differences. Fimbriae and LT enterotoxins are made up of large molecular weight proteins which facilitate their utilisation in vaccines and their detection using immunodiagnostic systems. The adhesive fimbriae and enterotoxins of animal ETEC are plasmid determined (except F41 and F17). Virulence gene probes (DNA hybridisation, PCR) are specific and sensitive diagnostic and epidemiologic tools for the detection of ETEC. Based on genetic typing, the ETEC, in spite of limited serogroups, seem to represent a population of E. coli with a diverse genetic background. PMID- 10367359 TI - Escherichia coli as a pathogen in dogs and cats. AB - Certain strains of Escherichia coli behave as pathogens in dogs and cats causing gastro-intestinal and extra-intestinal diseases. Among the five known groups of diarrhoeagenic E. coli, namely enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), shiga-toxin producing E. coli (STEC) and enteroaggregative E. coli (EAggEC), only EPEC and ETEC were clearly associated with enteric disease in young dogs. ETEC isolates from diarrhoeic dogs were found to be positive for the heat-stable enterotoxins STa and STb but negative for heat-labile enterotoxin (LT). Canine ETEC were found to be different from those of other animals and humans by their serotypes, production of alpha haemolysin and adhesive factors and by the production of uncharacterized types of enterotoxins by some ETEC. Canine EPEC could be distinguished from EPEC of humans or other animals by their serotypes and by the eae-protein intimin which mediates intimate adherence of EPEC to intestinal mucosa cells. STEC were occasionally isolated from faeces of healthy and diarrhoeic dogs but their role in canine diarrhoea is not yet well known. EIEC and EAggEC were not reported to occur in dogs or cats. Very little is known on diarrhoegenic E. coli in cats and further epidemiological investigations on this subject are needed. Besides its role in gastro-intestinal infections, E. coli can cause infections of the urogenital tract and systemic disease in dogs and cats. Extra-intestinal pathogenic E. coli strains from dogs and cats belong to a limited number of serotypes and clonal groups and are frequently found as a part of the normal gut flora of these animals. Many of these E. coli strains carry P-fimbriae and produce alpha haemolysin and a necrotizing cytotoxin (CNF1). Some of the frequently isolated types of extra-intestinal pathogenic E. coli from dogs, cats and humans were found to be highly genetically related but showed differences in their P-fimbrial adhesins which determine host specificity. Transmission of extra-intestinal and enteral pathogenic E. coli between dogs and humans was reported. Further research is needed, however, to determine the role of dogs and cats as transmission vectors of pathogenic E. coli strains to other animals and humans. PMID- 10367360 TI - Avian pathogenic Escherichia coli (APEC). AB - Avian pathogenic Escherichia coli (APEC) cause aerosacculitis, polyserositis, septicemia and other mainly extraintestinal diseases in chickens, turkeys and other avian species. APEC are found in the intestinal microflora of healthy birds and most of the diseases associated with them are secondary to environmental and host predisposing factors. APEC isolates commonly belong to certain serogroups, O1, O2 and O78, and to a restricted number of clones. Several experimental models have been developed, permitting a more reliable evaluation of the pathogenicity of E. coli for chickens and turkeys. Hence, virulence factors identified on APEC are adhesins such as the F1 and P fimbriae, and curli, the aerobactin iron sequestering system, K1 capsule, temperature-sensitive hemagglutinin (Tsh), resistance to the bactericidal effects of serum and cytotoxic effects. Experimental infection studies have shown that the air-exchange regions of the lung and the airsacs are important sites of entry of E. coli into the bloodstream of birds during the initial stages of infection and that resistance to phagocytosis may be an important mechanism in the development of the disease. They have also demonstrated that F1 fimbriae are expressed in the respiratory tract, whereas P fimbriae are expressed in the internal organs of infected chickens. The role of these fimbrial adhesins in the development of disease is not yet, however, fully understood. The more recent use of genetic approaches for the identification of new virulence factors will greatly enhance our knowledge of APEC pathogenic mechanisms. Diagnosis of APEC infections is based on the clinical picture, lesions and isolation of E. coli. This may be strengthened by serotyping and identification of virulence factors using immunological or molecular methods such as DNA probes and PCR. Approaches for the prevention and control of APEC infections include the control of environmental contamination and environmental parameters such as humidity and ventilation. Antibiotherapy is widely used, although APEC are frequently resistant to a wide range of antibiotics. Vaccines containing killed or attenuated virulent bacteria protect against infection with the homologous strain but are less efficient against heterologous strains. Hence, vaccination for colibacillosis is not widely practised because of the large variety of serogroups involved in field outbreaks. PMID- 10367362 TI - Genitourinary trauma. PMID- 10367361 TI - AFA and F17 adhesins produced by pathogenic Escherichia coli strains in domestic animals. AB - AFA and F17 are afimbrial and fimbrial adhesins, respectively, produced by pathogenic Escherichia coli strains in domestic animals. F17-related fimbriae are mainly detected on bovine and ovine E. coli associated with diarrhoea or septicaemia. The F17-G adhesin subunits recognize N-acetyl-D-glucosamine (GlcNAc) receptors present on bovine intestinal cells. Some F17 subtypes also bind to GlcNAc receptors present on human uroepithelial and intestinal Caco-2 cells or to the laminin contained in the basement of mammalian membranes. F17 is often associated with other virulence factors (aerobactin, serum resistance, CNF2 toxin, K99, CS31A or AFA adhesins) on pathogenic E. coli. A cluster of only four genes is required to synthesize functional F17-related fimbrial structures. The hypothesis of multifunctional F17 fimbrial subunits is supported by the fact that: i) the N-terminal part of the adhesin subunit participates in receptor recognition, whereas the C-terminal part is required for biogenesis of the fimbrial filament; and ii) the interaction between structural and adhesin subunits seems to be crucial for the initiation of monomer polymerization. Recently, determinants related to the afa gene clusters from human pathogenic E. coli associated with intestinal and extra-intestinal infections were identified in strains isolated from calves and piglets with diarrhoea and septicaemia. Two afa-related gene clusters, designated afa-7 and afa-8, that encode afimbrial adhesins were cloned and characterized from bovine pathogenic E. coli. These animal afa gene clusters were plasmid and chromosome borne and were expressed by strains that produced other virulence factors such as CNF toxins, F17, PAP and CS31A adhesins. A high frequency of afa-8 and a low prevalence of afa-7 among bovine E. coli isolates were suggested by preliminary epidemiological studies. As with the human afa gene clusters, the animal ones encode an adhesive structure composed of two proteins: AfaE which mediates adhesion to epithelial cells and AfaD which is an invasin. PMID- 10367363 TI - Radiographic staging of renal injuries. AB - Radiographic staging of renal injuries is the orderly process of establishing an immediate diagnosis so as to expedite effective treatment. Adult patients with gross hematuria or microhematuria associated with shock should undergo urgent imaging. Computerized tomography (CT scan) is the study of choice for evaluation of stable adult and pediatric patients with suspected renal trauma. Those in need of immediate surgical intervention are best evaluated by one-shot intravenous pyelogram (IVP) to determine the extent of the injured kidney and to document the normal function of the contralateral unit. Successful management of renal trauma is guided to a major degree by appropriate renal imaging. PMID- 10367364 TI - Renal trauma: indications and techniques for surgical exploration. AB - Injury secondary to trauma has become increasingly common in modern society. In the United States, in excess of 55 million trauma patients are evaluated each year, and trauma is the leading cause of mortality in people under the age of 40 years. Of the patients with abdominal trauma, approximately 10% have an injury to the urinary tract. Renal injury, occurring in 1-5% of all traumas, is due primarily to blunt trauma. Advances in the imaging and staging of renal trauma as well as in treatment strategies have decreased the need for surgical intervention and increased renal preservation. Nevertheless, no consensus exists regarding indications and techniques for renal exploration. The goals of treatment include accurate staging, maximal preservation of renal function, and minimal complications. We discuss our current approach in the management of renal trauma. PMID- 10367365 TI - Current methods of diagnosis and management of ureteral injuries. AB - A delay in diagnosis is the most important contributory factor in morbidity related to ureteral injury. The difficulty in making the diagnosis can be minimized by maintenance of a high index of suspicion and the timely performance of the appropriate radiographic and intraoperative evaluations. A decision on the timing of repair of the ureteral injury is based on the patient's overall condition, promptness of injury recognition, and proper injury staging. Ideally, when identified promptly, ureteral injuries should be repaired immediately. However, once there has been a delay in diagnosis or in the case of an unstable patient, temporizing measures can be used for urinary diversion. With the availability of simple, minimally invasive techniques to manage urinary extravasation and the absence of any risk of ureteral hemorrhage, ureteral reconstruction can be safely deferred until an opportune time during the recovery period. Successful surgical management requires familiarity with the broad reconstructive armamentarium and meticulous attention to the specific details of each procedure. Through adherence to the diagnostic and therapeutic principles outlined, complications can be minimized and renal preservation can be maximized in patients sustaining ureteral injuries. PMID- 10367366 TI - Bladder rupture from external trauma: diagnosis and management. AB - Rupture of the bladder from external trauma is usually due to a blow to the abdomen when the bladder is distended or associated with a fractured pelvis. The diagnosis is made by performance of a retrograde cystogram and observation of contrast extravasation. An intravenous contrast study is not acceptable. All patients with an intraperitoneal bladder rupture should have formal repair. Extraperitoneal bladder ruptures may be treated with catheter drainage if the urine clears of blood promptly, the catheter drains well, and the bladder neck is not involved in the injury. Otherwise, formal repair is mandatory. PMID- 10367367 TI - Pelvic fracture injuries of the posterior urethra. AB - Posterior urethral injuries are not common and take a low priority in the management of patients with pelvic fracture injuries as these individuals almost always have multiple injuries of more serious consequence. Most patients are best treated by a suprapubic catheter for 3 months and then an end-to-end anastomotic urethroplasty in those who have developed urethral occlusions. There are roles for delayed primary repair and for endourological management in selected patients, but their exact roles have yet to be defined. PMID- 10367368 TI - Anterior urethral injury. AB - Anterior urethral injuries, although rare, may be associated with substantial long-term morbidity. Urethral injuries that occur due to penetrating trauma or penile fracture are best treated by meticulous two-layer primary repair. Those that occur due to rapid deceleration injury are usually best treated by suprapubic diversion and delayed reconstruction because of associated injuries and concomitant contusion of the supporting spongiosum. PMID- 10367369 TI - Penile fracture and testicular rupture. AB - Traumatic injuries to the penis and testicles are uncommon, likely due to the well-protected location and degree of mobility of these organs. Because of this the management of these injuries has historically been controversial. However, current literature supports immediate evaluation and surgical repair of these traumatic injuries to prevent complications such as erectile dysfunction or testicular loss. Herein the diagnostic and therapeutic options for both traumatic penile fracture and testicular rupture are reviewed with emphasis on immediate evaluation and repair. PMID- 10367370 TI - Genital skin loss: unified reconstructive approach to a heterogeneous entity. AB - Genital skin loss results from a heterogeneous group of traumatic and infectious processes. Removal of all devitalized tissue is necessary prior to reconstructive surgery; hyperbaric oxygen may help in the management of necrotizing infections. Defect coverage depends more on the remaining genital skin than on the etiology of the loss. PMID- 10367371 TI - The need for microdissectional tumor cell preparation during the molecular genetic analysis of prostate cancer. AB - For clinically localized prostate cancer, recent studies strongly indicate that the determination of p53 inactivation allows the identification of a highly aggressive subgroup of prostatic tumors associated with decreased recurrence-free and long-term survival following radical prostatectomy. However, several questions regarding the determination of p53 alterations in prostate cancer, such as the poor correlation between immunohistochemistry and molecular genetic analysis, remain to be clarified. On the DNA level, p53 gene alterations have been identified in only up to 64% of tumors exhibiting immunohistochemically detected overexpression of the p53 oncoprotein. This discrepancy can be explained either by the genetic microheterogeneity of prostate cancer or by stabilization of the wild-type protein due to posttranslational events. In the present study we tried to determine the concordance between an immunohistochemically detected p53 overexpression and the result of molecular genetic analysis. Therefore, tumor tissue obtained by microdissection from 40 prostate cancer specimens was subjected to DNA-sequence analysis. Microdissection was based either only on histopathologic criteria or on the result of the immunohistochemical staining reaction. In 8 of 14 (57%) tumors a positive immunohistochemical reaction could be confirmed by DNA sequencing, which revealed a missense point mutation at the p53 gene locus, mainly in the form of G-->A transversion in exon 5 of the p53 gene. Following the micropreparation of tumor cells exhibiting p53 oncoprotein overexpression, missense point mutation could be detected in an additional 4 cases. Following a microscopically guided tumor cell dissection according to the result of immunohistochemistry, DNA sequencing confirmed an immunohistochemically detected p53 overexpression in 86% of cases investigated. This result indicates that a microdissectional tumor cell preparation is recommended for molecular genetic analysis of histologically heterogeneous tissue specimens such as prostate cancer and should be performed according to and in addition to the result of immunohistochemistry when an immunohistochemical approach is available. PMID- 10367372 TI - Angiomyolipoma of the kidney and lymph nodes. AB - Angiomyolipoma (AML) is a benign mesenchymal tumor predominantly occurring in the kidney. Despite its low incidence of 0.07-0.03% in an unselected population, this tumor is well known, because the typical AML can be diagnosed without histological confirmation by a combination of ultrasound (US) and computerized tomography (CT) imaging in up to 95% of cases. In contrast, simultaneous involvement of the kidney and the regional lymph nodes is less known and might be confused with metastasizing malignant tumor. We report a case of the very uncommon simultaneous involvement of the kidney and the lymph nodes in AML. PMID- 10367373 TI - Positive bladder cooling reflex in patients with bladder outlet obstruction due to benign prostatic hyperplasia. AB - The bladder cooling reflex was evaluated in patients with bladder outlet obstruction to study the effect of obstruction on the afferent neural function of the bladder, especially on the C-afferents. The bladder cooling test was performed by infusion of 0 degree C saline into the bladder with simultaneous detrusor pressure measurement in 104 patients with bladder outlet obstruction due to benign prostatic hyperplasia. In 49 patients (47%) a positive cooling reflex was observed. This was defined as a rise in the detrusor pressure following cold saline instillation exceeding 15 cmH2O (range 15-130 cmH2O, mean 60.6 cmH2O; positive group). In the remainder of cases the pressure rise ranged from 0 to 12 cmH2O (mean 6.1 cmH2O; negative group). Bladder outlet obstruction may cause some alteration in the afferent neural function of the bladder, in particular of the C afferent fibers. PMID- 10367374 TI - Pesticide residue analysis: 1997-1998. PMID- 10367375 TI - Use of microwave digestion and atomic absorption spectrophotometry to determine chromic oxide as a digestibility marker in feed, feces, and ileal content. AB - Most conventional digestion procedures, such as dry ashing and wet ashing, are tedious and labor intensive. Microwave digestion is a good alternative, because microwave dissolution is faster, safer, and simpler, and provides more controlled reproducible conditions than conventional methods. The purpose of this study was to develop a microwave digestion method for mineralizing meat and bone meal diets, feces, and ileal contents. Each sample was heated on a hot plate for 10 min, dry ashed at 65 degrees C for 4 h, and transferred into microwave vessels. Then, 10 mL 70% HNO3 was added. Samples were digested for 7, 10, and 20 min at 95, 90, and 85% power, respectively. After the heating cycle, 6 mL 30% H2O2 was added, and samples were returned to the microwave for a second heating cycle of 1 and 7 min at 95% and 90% power, respectively. Finally, chromium concentration was determined by flame atomic absorption spectrophotometry. The digestion method was validated by using a standard reference material, SRM domestic sludge 2781, with a certified chromium value of 195 +/- 9 micrograms/g. The value obtained in this study was 178 +/- 11 micrograms/g, for a difference of 17 micrograms/g. Spike recovery experiments resulted in 103.16 and 100.35% recoveries of chromium from diet and feces samples, respectively. Coefficients of variation were 10.8 and 7.8%, respectively. PMID- 10367376 TI - Determination of avilamycin in poultry feeds by liquid chromatography. AB - Avilamycin was extracted from feed with acetonitrile. Isolation of avilamycin factors from feed matrix interference was accomplished by normal-phase solid phase extraction with silica as sorbent. Reversed-phase liquid chromatography was subsequently used to separate and quantitate the primary biologically active factors A and B for determination of chemical potency. This method combines specificity for avilamycins A and B in poultry feeds with simple sample preparation that removes matrix interferences. Recoveries of factor A ranged from 93.29 to 97.26%, with precision (relative standard deviation) ranging from 1.1 to 3.4%. Avilamycin factors in feed samples tested ranged from 4.45 to 17.82 micrograms/g for factor A and from 0.80 to 3.18 micrograms/g for factor B. PMID- 10367377 TI - Stability-indicating methods for determining omeprazole and octylonium bromide in the presence of their degradation products. AB - Four stability-indicating assays were developed for determining omeprazole and octylonium bromide. Omeprazole is photodegraded and estimated in the presence of its degradation products sulphenamide (I) and benzimidazole sulphide (II) by 2 methods. The first method depends on use of first-, second-, and third-derivative spectrophotometry at 290.4, 320.6, and 311.6 nm, respectively. The second method is based on applying the charge-transfer technique with chloranil as pi acceptor to form a complex with omeprazole, the absorbance of which is measured at 377 nm. These methods determine omeprazole in concentration ranges of 5-20 micrograms/mL by first-, second-, and third-derivative spectrophotometry and 10-50 micrograms/mL by charge-transfer complexation with mean accuracies of 99.92 +/- 0.73, 99.71 +/- 1.02, 99.64 +/- 0.66, and 100.24 +/- 0.81%, respectively. Octylonium bromide is determined by a densitometric method using thin-layer chromatography in the presence of its degradation products p-[2-(n octyoxy)benzoyl]-aminobenzoic acid (III) and diethyl-(2-hydroxyethyl)-methyl ammonium bromide (IV) without any interferences. Alternatively, octylonium bromide is evaluated by a colorimetric method using the acid dye rose bengal. The ion pair formed is extracted in chloroform at pH 4, and its absorbance is measured at 562 nm. These methods determine octylonium bromide in the presence of its degradation products in concentration ranges of 0.1-0.5 microgram/microL by densitometry and 4.5-22.5 micrograms/mL by colorimetry, with mean accuracies of 100.21 +/- 0.93 and 99.73 +/- 0.89%, respectively. The suggested methods were used to determine drugs in bulk powder, laboratory-prepared mixtures, and pharmaceutical dosage forms. Results were compared statistically with those obtained with reference methods. PMID- 10367378 TI - Determination of four fluoroquinolones in milk by on-line immunoaffinity capture coupled with reversed-phase liquid chromatography. AB - An automated, on-line immunoaffinity extraction method was developed for the analysis of 4 fluoroquinolones in milk: ciprofloxacin, difloxacin, enrofloxacin, and sarafloxacin. This method involves analyte extraction using an immunoaffinity capture column containing anti-fluoroquinolone antibodies coupled on-line with reversed-phase column chromatography. Liquid chromatographic analyses were performed by isocratic elution using a mobile phase of 2% acetic acid acetonitrile (85 + 15) and an Inertsil phenyl column with fluorescence detection at excitation and emission wavelengths of 278 and 444 nm, respectively. No significant interferences from the sample matrix were observed, indicating good selectivity with the immunoaffinity column. Recoveries from fortified raw milk samples (5-50 ppb of each fluoroquinolone) ranged from 72 to 90%, with standard deviations of < or = 8%. PMID- 10367380 TI - Gas chromatographic analysis of fosfomycin in plasma for pharmacokinetic analysis. AB - An efficient method for gas chromatographic analysis of fosfomycin in plasma was developed for preliminary investigations of the bioavailability in poultry of 3 commercial complexes of fosfomycin: a levorotatory Ca(-) salt, a racemic Ca(+/-) salt, and a tromethamine (THAM) salt. The method was used to determine whether the less expensive racemic mixture would provide equivalent levels of fosfomycin in blood as the pure Ca(-) form and the THAM salt. The THAM salt, a more expensive product to market, was thought to have the greatest bioavailability. The assay is selective, sensitive, and applicable to pharmacokinetic analysis. PMID- 10367379 TI - Determination of flumequine in channel catfish by liquid chromatography with fluorescence detection. AB - Rapid methods are described for determination of flumequine (FLU) residues in muscle and plasma of farm-raised channel catfish (Ictalurus punctatus). FLU residues were extracted from tissues with an acidified methanol solution, and extracts were cleaned up on C18 solid-phase extraction cartridges. FLU concentrations were determined by liquid chromatography (LC) using a C18 analytical column and fluorescence detection (excitation, 325 nm; emission, 360 nm). Mean recoveries of FLU from fortified muscle were 87-94% at 5 levels ranging from 10 to 160 ppb (5 replicates per level). FLU recoveries from fortified plasma were 92-97% at 5 levels ranging from 20 to 320 ppb. Limits of detection (signal to-noise ratio, 3:1) for the method as described were 3 and 6 ppb for muscle and plasma, respectively. Relative standard deviations (RSDs) for recoveries were < or = 12%. Live catfish were dosed with 14C-labeled or unlabeled FLU to generate incurred residues. Recoveries of 14C residues throughout extraction and cleanup were 90 and 94% for muscle and plasma, respectively. RSDs for incurred FLU at 2 levels in muscle and plasma ranged from 2 to 6%. The identity of FLU in incurred tissues was confirmed by LC/mass spectrometry. PMID- 10367381 TI - Reveal for Salmonella test system. AB - The Reveal for Salmonella (RSS) test system is a presumptive qualitative test that detects the presence of Salmonella organisms in foods within 21 h total testing time, allowing the user to release negative products 24 h earlier than when using other rapid test kits. Foods are enriched with a proprietary resuscitation medium called Revive and then selectively enriched with either Selenite Cystine or Rappaport-Vassiliadis selective media. The enriched culture is used to inoculate the RSS detection device, which initiates a lateral flow through a reagent zone containing anti-Salmonella antibodies conjugated to colloidal gold particles that capture antigens present in the culture. The antigen-antibody complex migrates farther and is captured by an additional anti Salmonella antibody, causing the colloidal gold to precipitate and form a visual line, indicating a positive result. A procedural control line also will form regardless of the presence of Salmonella organisms to indicate the test is working properly. Existing AOAC Official Methods for Salmonella organisms require a 48 h enrichment before testing. Hence, a food product has to be held before release, adding extra cost to the company and the consumer. The RSS test system was evaluated by quantitative spiking studies. Although AOAC encourages inclusion of naturally contaminated foods, almost all microbiological AOAC validation studies have been performed with artificially contaminated foods for absolute control over the study. The RSS test system is designed to test many food types for Salmonella organisms and has a limit of detection of 5-10 colony-forming units (cfu)/25 g with a false-negative rate of < 1% and a false-positive rate of < 5.0%. It showed an 81% overall agreement with the traditional procedure of the U.S. Department of Agriculture's Food Safety Inspection Service. PMID- 10367382 TI - Salmonella in foods--a new enrichment procedure for use with the TECRA Salmonella visual immunoassay: collaborative study. AB - A collaborative study was conducted to compare a new enrichment procedure for the TECRA Salmonella Visual Immunoassay with the reference method given in the U.S. Food and Drug Administration's Bacteriological Analytical Manual (BAM 7th Ed.). Three food types (milk powder, black pepper, and soy flour) were analyzed in Australia, and 3 food types (milk chocolate, dried egg, and raw turkey) were analyzed in the United States. Thirty-eight collaborators participated in the study. No significant differences (p > 0.05) were observed for the pairwise comparison of the proportion of positive samples for the TECRA method with that for the reference method. The new enrichment procedure for the TECRA method has been adopted First Action by AOAC INTERNATIONAL. PMID- 10367383 TI - Microbial ranking of porous packaging materials (exposure chamber method), ASTM method: collaborative study. AB - A collaborative study involving 11 laboratories was conducted to measure the microbial barrier effectiveness of porous medical packaging. Two randomly cut samples from each of 6 commercially available porous materials and one positive and one negative control were tested by one operator in each of 11 laboratories. Microbial barrier effectiveness was measured in terms of logarithm reduction value (LRV), which reflects the log10 microbial penetration of the material being tested. The logarithm of the final concentration is subtracted from that of the initial concentration to obtain the LRV. Thus the higher the LRV, the better the barrier. Repeatability standard deviations ranged from 6.42 to 16.40; reproducibility standard deviations ranged from 15.50 to 22.70. Materials B(53), C(50), D(CT), and E(45MF) differ significantly from the positive control. The microbial ranking of porous packaging materials (exposure chamber method), ASTM method, has been adopted First Action by AOAC INTERNATIONAL. PMID- 10367384 TI - Fusarium mycotoxins in corn and corn products in a high-risk area for gastric cancer in Shandong Province, China. AB - Consumption of fermented, but not unfermented, corn pancakes has been linked with elevated stomach cancer mortality rates in rural Linqu County in Shandong Province, China. Previous surveys of fungal contamination of corn in China have detected fumonisins, which are mycotoxins produced by Fusarium moniliforme. To determine whether mycotoxins might account for the increased risk of cancer among those consuming fermented pancakes, we obtained specimens of corn, cornmeal, unfermented and fermented pancake batter, and cooked fermented pancakes from each of 16 households in Linqu County for analysis by the U.S. Department of Agriculture. Fumonisins B1, B2, and B3 were detected (> or = 0.5 microgram/g) in 19, 25, and 6% of the corn specimens, respectively, as well as in various corn products. No type A trichothecenes were detected; however, the type B trichothecenes deoxynivalenol and 15-acetyldeoxynivalenol were detected (> or = 0.5 microgram/g) in 58 and 17% of the corn specimens, respectively, and zearalenone was detected (> or = 0.5 microgram/g) in 15% of the cornmeal specimens. The mycotoxins were detected only at low levels (< 10 micrograms/g), which did not increase with fermentation. These findings do not support the hypothesis that mycotoxin contamination increases the risk of gastric cancer among those who consume fermented Chinese pancakes. PMID- 10367385 TI - Analysis of beta-carotene in medical food by liquid chromatography with matrix solid-phase dispersion. AB - A liquid chromatographic method is described for analysis of beta-carotene in medical food. The nutrient is extracted from medical food without saponification by matrix solid-phase dispersion and quantitated by isocratic normal-phase chromatography with a Si 60 column and a mobile phase of hexane containing 0.125% (v/v) isopropyl alcohol. The limit of quantitation is 0.02 microgram/mL at 436 nm. Standard response was linear over the concentration range of 0.02-1.0 microgram/mL (r2 = 0.99998). Recoveries were determined on a zero control reference material containing added beta-carotene at various levels. Recoveries averaged 91.2% (n = 25) with coefficients of variation from 0.50 to 3.10%. The method provides a rapid, specific, sensitive, and easily controlled assay for analysis of beta-carotene in fortified medical food. In addition, retinyl palmitate can be assayed simultaneously with an in-line fluorescence detector. PMID- 10367386 TI - An immunoaffinity column for the determination of peanut protein in chocolate. AB - An immunoaffinity column was prepared from rabbit polyclonal antiserum for the determination of peanut protein from food matrixes. The anti-peanut immunoglobulin G was isolated from antiserum by affinity chromatography on a column coupled with peanut protein and then attached to an AminoLink gel. The column was applied to the determination of peanut protein in chocolate after extraction, immunoaffinity chromatography, and enzyme-linked immunosorbent assay (ELISA). Overall recoveries from chocolate spiked with 0.2-3.2 micrograms/g of peanut protein averaged 77% (range, 72-84%), and the minimum detection limit was 0.1 microgram/g. Chromatography of extracts with the column improved detection limit and eliminated the matrix effect experienced with direct ELISA of chocolate extracts. PMID- 10367387 TI - Principles to guide international standard tests for liquid chemical germicides: a proposal. AB - This review discusses issues involved in developing standard tests for liquid chemical germicides and suggests some guiding principles to be considered for future development of harmonized international standard methods for testing disinfectants and sterilants. A published test method to measure sporicidal activity is used as an example of the implementation of these principles. PMID- 10367388 TI - Dietary intake and residues of organochlorine pesticides in foods from Hsinchu, Taiwan. AB - The levels of contamination with various organochlorine pesticides (such as total HCH, heptachlor, heptachlor epoxide, aldrin, dieldrin, endrin, endosulfan, and total DDT) of different foods from 3 traditional markets were determined to estimate Taiwanese daily intake of organochlorine pesticides. Of the 18 organochlorine pesticides investigated, alpha-HCH, beta-HCH, lindane, delta-HCH, heptachlor, heptachlor epoxide, dieldrin, endrin, alpha-endosulfan, p,p'-DDE, and p,p'-DDT were detected at concentrations ranging from 0.26 to 10.2 ng/g wet weight. Contamination with organochlorine pesticides followed the order heptachlor > dieldrin > alpha-endosulfan > HCH isomers > heptachlor epoxide > DDT. Frequencies of detection of organochlorine pesticide residues ranged from 2.0 to 52.3%. alpha-Endosulfan was the most frequently detected organochlorine pesticide in the foods analyzed, followed by heptachlor epoxide (47.6%) and alpha HCH (38.9%). Estimated daily intakes (EDIs) of organochlorine pesticides from foods were 1.137 micrograms for total HCH, 2.147 micrograms for heptachlor, 0.702 microgram for heptachlor epoxide, 0.624 microgram for endosulfan, 0.098 microgram for cyclodiene, and 0.541 microgram for total DDT. These EDIs were only 0.075% of the acceptable daily intake (ADI) for lindane, 47.5% of ADI for heptachlor and heptachlor epoxide, 0.045% of ADI for total DDT, and 1.01% of ADI for aldrin and dieldrin. Therefore, consumption of the foods analyzed does not pose a risk to consumer health. PMID- 10367389 TI - Determination of haloacetic acids in water by ion chromatography--method development. AB - The microextraction/ion chromatographic (IC) method developed in this study involves extraction of 9 haloacetic acids (HAAs) from aqueous samples (acidified with sulfuric acid to a pH of < 0.5 and amended with copper sulfate pentahydrate and sodium sulfate) with methyl tert-butyl ether (MTBE), back extraction into reagent water, and analysis by IC with conductivity detection. The separation column consists of an Ion Pac AG-11 (2 mm id x 50 mm length) guard column and an Ion Pac AS-11 (2 mm id x 250 mm length) analytical column, and the concentration column is a 4 mm id x 35 mm length Dionex TAC-LP column. Use of the 2 mm id Dionex AS-11 column improved detection limits especially for trichloracetic acid (TCAA), bromodichloroacetic acid (BDCAA), dibromochloroacetic acid (DBCAA), and tribromoacetic acid (TBAA). The peak interfering with BCAA elutes at the same retention time as nitrate; however, we have not confirmed the presence of nitrate. Stability studies indicate that HAAs are stable in water for at least 8 days when preserved with ammonium chloride at 100 mg/L and stored at 4 degrees C in the dark. At day 30, recoveries were still high (e.g., 92.1-106%) for dichloroacetic acid (DCAA), BCAA, dibromoacetic acid (DBAA), trichloroacetic acid (TCAA), BDCAA, and DBCAA. However, recoveries of monochloroacetic acid (MCAA), monobromoacetic acid (MBAA), and TBAA were only 54.6, 56.8, and 66.8%, respectively. Stability studies of HAAs in H2SO4-saturated MTBE indicate that all compounds except TBAA are stable for 48 h when stored at 4 degrees C in the dark. TBAA recoveries dropped to 47.1% after 6 h of storage and no TBAA was detected after 48 h of storage. The method described here is only preliminary and was tested in only one laboratory. Additional research is needed to improve method performance. PMID- 10367390 TI - Maternal body burden of organochlorine pesticides and dioxins. AB - To investigate the body burden of organochlorine pesticides and dioxins in Japanese women, 125 milk samples were collected from 41 mothers in 1994, 42 in 1995, and 42 in 1996. Of the 125 samples, 82 were from primipara mothers (first delivery) and 43 were from multipara mothers (second or later delivery). By using capillary gas chromatography with electron capture detection, beta-HCH and p,p' DDE were detected as the major chlorine pesticides in human milk. Average levels of beta-HCH and p,p'-DDE were 475 and 368 ng/g lipid, respectively, in primipara breast milk, 314 and 259 ng/g lipid in multipara breast milk, and 420 and 330 ng/g lipid in total breast milk. Dieldrin, heptachor epoxide, oxychlordane, trans chlordane, and cis-chlordane were detected at lower average levels of 3, 4, 34, 41, and 5 ng/g lipid, respectively. By using high-resolution gas chromatography with mass spectrometric detection, dioxins were detected in all samples. Average levels of total polychlorinated dibenzo-p-dioxin (PCDD), total polychlorinated dibenzofuran (PCDF), total PCDD + PCDF, total coplanar polychlorinatedbiphenyl (CoPCB), and total dioxin were 10.0, 7.8, 17.7, 9.9, and 27.5 TEQ (toxic equivalent) pg/g lipid, respectively, in primipara breast milk; 7.0, 5.8, 12.8, 7.3, and 20.1 TEQ pg/g lipid in multipara breast milk; and 8.9, 7.1, 16.1, 8.9, and 25.0 TEQ pg/g lipid in total breast milk. In primipara breast milk, significant correlations were found among levels of beta-HCH, p,p'-DDE, total PCDD-TEQ, total PCDF-TEQ, total CoPCB-TEQ, and total TEQ except for less correlation between p,p'-DDE and total PCDF-TEQ. Levels of these analytes also significantly increased depending on mother's age, except for total Co-PCB-TEQ. For the correlation with food habit, the only positive correlation was between total PCDF-TEQs and fish intake. PMID- 10367391 TI - The impact of household preparations on the residues of pesticides in selected agricultural food commodities available in India. AB - Gas chromatographic multiresidue methods for simultaneous determination of organophosphorus, organochlorine, and organonitrogen pesticides were used to study the exposure of the Indian population to pesticide contamination at their actual dietary intakes. Selected agricultural commodities--5 kinds of vegetables (tomato, potato, okra, cabbage, and green beans), 6 kinds of cereals and pulses (rice, maize, wheat, red gram, black gram, and green gram), and 6 kinds of fruits (mango, orange, guava, banana, apple, and grapes)--readily available in Chennai City local markets--were studied for this purpose. Samples were fortified with known concentrations of various pesticides and subjected to household preparation methods commonly used in India. The impact of household preparation is very high, resulting in 65-95% decontamination of pesticides at different stages. Of 512 raw market samples analyzed, the organochlorine and organophosphorus pesticides present in 12 samples were removed during household preparations, resulting in residues well below the toxicologically acceptable limits. PMID- 10367392 TI - Determination of fat, protein, and total solids in ovine milk by near-infrared spectroscopy. AB - Analysis by near-infrared spectroscopy (NIRS) was investigated as a means of predicting quality parameters of ovine milk. Calibration equations were developed with samples of ovine milk obtained from a flock of Manchega and Lacaune dairy ewes at different stages of lactation for a wide variation in milk composition. Prediction equations for milk protein, fat, and total solids content were developed by use of reflection or transflection methods to measure absorbance values. Accuracies of measurements were compared. R2 (squared multiple correlation coefficient) values were satisfactory in most cases. The highest R2 value for milk protein content (0.92) was obtained in transflectance mode with unhomogenized milk. The highest R2 values for fat (0.99) and total solids (0.98 0.96) content were obtained in both a transflectance mode without sample conditioning and in a transflectance mode with milk homogenized at 40 degrees C. To validate the calibration, an independent set of 40 milk samples was used. The best r2 (simple correlation coefficient) values for protein, fat, and total solids were 0.92, 0.97, and 0.92, respectively. The study showed that NIRS is a potentially useful technique for evaluating the composition of unhomogenized ovine milk. PMID- 10367393 TI - Measurement of sugars and starches in foods by a modification of the AOAC total dietary fiber method. AB - A separation scheme for the determination of sugars and starch in processed food was developed. It is based on AOAC Method 985.29 for total dietary fiber with these modifications: carbohydrate starches are separated into soluble and insoluble fractions before they are hydrolyzed; acetonitrile is used instead of ethanol to separate sugars from enzyme-resistant carbohydrates, proteins, and other macromolecules; and a solid-phase extraction filter is included to remove substances that interfere with high-performance liquid chromatography (HPLC). Recovery studies indicate a > 97% sugar recovery. Twenty foods were analyzed. After enzymatic hydrolysis, fructose, glucose, sucrose, maltose, and lactose were extracted and determined by HPLC using a refractive index detector. Starch content was calculated from the increase in the amount of glucose. The results were compared with values listed on the "Nutrition Facts" panel for that food. The analyzed amounts of sugars and starches were 73-96% of declared values. PMID- 10367394 TI - Rapid and simple determination of oxytetracycline in chicken products. AB - A rapid and simple method for determining residual oxytetracycline (OTC) in chicken products (muscle, liver, and eggs) by high-performance liquid chromatography (HPLC) was developed. Samples were prepared by homogenization with acetonitrile-n-hexane (5 + 4, v/v) followed by centrifugation to minimize fat and protein contents. OTC in the acetonitrile layer was free from interfering compounds when examined by HPLC using a LiChrospher 100 RP-8 end-capped column, a mobile phase of acetonitrile-acetic acid-0.01 M disodium EDTA (28 + 2 + 70, v/v/v), and a photodiode array detector. Average recoveries from samples spiked with OTC (0.1, 0.2, and 1.0 ppm) were > 88%, with coefficients of variation between 2.3 and 5.1%. The limit of detection was 0.05 ppm. PMID- 10367395 TI - Phycotoxins. AB - The 1997-1998 period brought many new developments to the phycotoxin field. There were several reviews on phycotoxins in general, on their toxicological evaluation, and on their analysis. The ecophysiology, biosynthesis, and metabolism of polyether toxins and paralytic shellfish poisoning (PSP) toxins were also reviewed. The proceedings of the Eighth International Conference on Harmful Algae (Vigo, Spain, June 25-29, 1997) have been published and provide an excellent source of information on phycotoxins and toxic plankton bloom research. In addition, the much anticipated proceedings of the IX International IUPAC Symposium on Mycotoxins and Phycotoxins (Rome, Italy, May 27-31, 1996) have been published. Further evidence was provided to support the theory that Prorocentrum lima is the source organism for diarrhetic shellfish poisoning (DSP) toxins in Nova Scotian shellfish. In another study, different Prorocentrum species and isolates were analyzed for DSP toxins. In addition to detecting some new compounds, such as a DTX1 isomer, it was found that toxins were produced by both axenic and nonaxenic batch cultures, indicating that bacteria are probably not involved in the biosynthesis. The source organism for the spirolides, a family of fast-acting toxins reported from Nova Scotia, Canada, was determined to be Alexandrium ostenfeldii, a species that is found worldwide. The biogenetic origin of yessotoxin was reported to be Protoceratium reticulatum, another widely occurring organism. A great deal of attention and research funding has been directed at the serious problems associated with Pfiesteria piscicida. Analysts are eagerly awaiting publication of toxin structures, which will then allow the development of analytical methods. An incident of the mass mortality of California sea lions was reported in the Monterey area in May 1998. Analyses of tissue and urine samples revealed the presence of domoic acid. High levels of domoic acid were also found in anchovies and sardines, a common food source of sea lions. This is reminiscent of an incident of mass bird mortality in 1992 in the same region. Toxicological studies of domoic acid continue with one investigation on the effect of pH on toxicity in the mouse assay and others examining toxic effects in rats and cynomolgus monkeys. A study on the uptake and depuration of domoic acid in the Dungeness crab was reported. On October 20, 1997, EU (European Union) directive CE97/61 established a regulatory limit of 20 ppm for domoic acid in European shellfish, the same level as in North America. A detailed study on the oral toxicity of DSP toxins in mice was reported. Recent work by several researchers has revealed the genotoxic potential of okadaic acid and other DSP toxins. Previous work had clearly demonstrated the tumor-promoting potential of DSP toxins, but this recent evidence, which shows mutations in the progeny of okadaic acid-treated cells and the formation of DNA-adducts, increases concerns over the hazards associated with DSP-contaminated shellfish. The toxicology of yessotoxin was evaluated by Ogino et al. The toxin showed weak cytotoxicity, but was not orally lethal to mice at 10 mg/kg, and did not cause intestinal fluid accumulation, inhibition of protein phosphatase 2A (PP2A), or hemolytic effects. Similarly, Tubaro et al. saw no evidence for diarrheogenicity of homoyessotoxin isolated from mussels and from the proposed planktonic producer, Lingulodinium polyedrum. All this provides further evidence that yessotoxin should not be classed as a DSP toxin. A number of new toxins have been detected and identified. Two analogues of yessotoxin, homoyessotoxin, and 45 hydroxyhomoyessotoxin were isolated from mussels of the Adriatic Sea and identified by Satake et al. A recent DSP event in Ireland associated with cultured mussels led to the identification of azaspiracid, a unique marine toxin with spiro ring assemblies. (ABSTRACT TRUNCATED) PMID- 10367396 TI - Plant toxins. PMID- 10367397 TI - Morphology of the pupal heart, adult heart, and associated tissues in the fruit fly, Drosophila melanogaster. AB - The early pupal heart of the fruit fly Drosophila melanogaster has recently been the subject of intense physiological and molecular work, yet it has not been well described, nor has it been compared with the heart of the adult fly. In the work reported here, the hearts of adults and early pupae of D. melanogaster were studied by scanning and transmission electron microscopy and by light microscopy. The hearts of adults and early pupae both consist of a tube of circular striated muscle one cell in thickness. The alary muscles, which suspend the heart, are more delicate in the adult compared to the early pupa. The pericardial cells in both early pupae and adults are connected to the heart by connective tissue radiating from the alary muscles or dorsal diaphragm. We confirm that four major changes occur in the heart during metamorphosis: 1) a conical chamber is formed de novo in the first and second abdominal segments; 2) the adult heart curves to conform to the contour of the abdomen; 3) a layer of longitudinal striated muscle appears on the ventral surface of the heart; 4) a fourth pair of ostia is added to the three already present in the early pupa; and note additionally that 5) the ostia appear as simple openings in the heart of the early pupa but are valve-like in the adult. PMID- 10367398 TI - Experimental alteration of limb posture in the chicken (Gallus gallus) and its bearing on the use of birds as analogs for dinosaur locomotion. AB - Extant birds represent the only diverse living bipeds, and can be informative for investigations into the life-history parameters of their extinct dinosaurian relatives. However, morphological changes that occurred during early avian evolution, including the unique adoption of a nearly horizontal femoral orientation associated with a shift in center of mass (CM), suggest that caution is warranted in the use of birds as analogs for nonavian dinosaur locomotion. In this study, we fitted a group of white leghorn chickens (Gallus gallus) with a weight suspended posterior to the hip in order to examine the effects on loading and morphology. This caused a CM shift that necessitated a change in femoral posture (by 35 degrees towards the horizontal, P < 0.001), and resulted in reorientation of the ground reaction force (GRF) vector relative to the femur (from 41 degrees to 82 degrees, P < 0.001). Despite similar strain magnitudes, an overall increase in torsion relative to bending (from 1.70 to 1.95 times bending, P < 0.001) was observed, which was weakly associated with a tendency for increased femoral cross-sectional dimensions (P = 0.1). We suggest that a relative increase in torsion is consistent with a change in femoral posture towards the horizontal, since this change increases the degree to which the bone axis and the GRF vector produce mediolateral long-axis rotation of the bone. These results support the hypothesis that a postural change during early avian evolution could underlie the allometric differences seen between bird and nonavian dinosaur femora by requiring more robust femoral dimensions in birds due to an increase in torsion. PMID- 10367399 TI - Biological assay for the detection of metallo-beta-lactamases in Bacteroides fragilis. AB - A biological assay was developed for the detection of carbapenemases, particularly metallo-beta-lactamases, in Bacteroides fragilis. The isolates tested possessed the gene (cfiA) responsible for metallo-beta-lactamase production, and showed reduced susceptibility to imipenem. Carbapenemase activity was investigated spectrophotometrically and by a biological assay in which sonicates of bacterial cells were mixed with imipenem in wells cut into Isosensitest agar inoculated with an Escherichia coli indicator organism. After incubation, zones of inhibition were measured. Reductions in zone size compared to a beta-lactamase-negative control, indicating carbapenemase production, were observed with all strains that exhibited hydrolysis of imipenem when measured spectrophotometrically, and with one isolate in which activity was not detected by spectrophotometry. Inclusion of EDTA in the well mixtures abolished the reduction in zone size, indicating the presence of metallo-beta-lactamase. This simple method can detect weak carbapenemase activity that may be overlooked by spectrophotometry. PMID- 10367400 TI - Incidence of Dientamoeba fragilis in faecal samples submitted for routine microbiological analysis. AB - Over a six-month period, 857 faecal samples were submitted to the Department of Microbiology and Immunology at Sultan Qaboos University Hospital in Oman, for routine microbiological examination. All samples were stained using the Gomori trichrome method. Trophozoites of Dientamoeba fragilis were detected in 41 (5.1%) patients, making it the most common enteropathogen found in the study. Of the patients with pure D. fragilis infection, 83% had abdominal pain, the duration of which varied from one month to two years. The use of permanently stained smears allowed detection of D. fragilis for the first time in the Sultanate of Oman. PMID- 10367401 TI - Seroprevalence of Helicobacter pylori infection in patients with hepatitis B. AB - Helicobacter pylori infection has been investigated extensively in immunocompromised hosts, such as those with acquired immunodeficiency syndrome (AIDS) and organ transplant recipients. However, few reports on H. pylori prevalence in individuals with chronic HBV infection are available. The aim of this serological study is to investigate H. pylori prevalence in patients with hepatitis B. Ninety-six consecutive hospitalised patients with chronic hepatitis B were studied, together with 104 age-matched healthy individuals of similar socioeconomic status and with no evidence of hepatitis B virus infection or liver diseases. Serum samples from both groups were tested for specific IgG antibodies to H. pylori, using enzyme-linked immunosorbent assay (ELISA). Of the 96 patients with hepatitis B, 55 (57.3%) were positive for serum IgG anti-H. pylori, significantly greater than in the control group of 104, where 44 (42.3%) were positive (P < 0.05). In addition, the seroprevalence of H. pylori in the 45 patients who were positive for hepatitis B envelope antigen (HBeAg) and/or HBV DNA was 75.6% (34), compared to 41.2% (21) in the 51 patients who were negative (P < 0.005). An increase in H. pylori prevalence is present in patients with chronic hepatitis B. Further study is needed to determine whether eradication of H. pylori will benefit these patients. PMID- 10367402 TI - Serovar distribution of Chlamydia trachomatis isolates collected from the cervix: use of the polymerase chain reaction and restriction endonuclease digestion. AB - In order to study the distribution of Chlamydia trachomatis serovars isolated from the cervix of patients attending the gynaecology out-patients clinic of Safdarjang Hospital, New Delhi, India, gene typing was performed using restriction fragment length polymorphism (RFLP) analysis of a polymerase chain reaction (PCR)-amplified portion of the major outer membrane protein (MOMP). A set of primers were used to amplify a 540 bp gene fragment which encompasses the four hypervariable regions of the MOMP. EcoR1 and Xbal double digestion of the product gave distinctive patterns for the genital serovars (D-K) as demonstrated on 12% polyacrylamide gel stained with ethidium bromide. PCR and RFLP were used to genotype 50 clinical isolates and their respective control serovars. Clinical isolates demonstrated the same banding pattern as the control strain of C. trachomatis. The serovars isolated were D (39.13%), E (28.26%), G (15.25%), I (10.86%) and F (6.5%), representing 92% of those investigated. PMID- 10367403 TI - Differential effects of sodium butyrate on the transcription of the human TIS11 family of early-response genes in colorectal cancer cells. AB - The tetradecanoyl phorbol acetate (TPA)-inducible sequence 11 (TIS11) family of early-response proteins consists of at least five members. They share a highly conserved Cys3His zinc-binding motif, but otherwise have little sequence similarity. Their function remains unknown, but all are induced rapidly and transiently in response to extracellular hormone and growth factor signals. Sodium butyrate, a fermentation product of dietary fibre, effects colorectal cancer cell proliferation by inducing growth arrest, differentiation and apoptosis. In this communication, we report that butyrate has differential effects on the transcription of the three human TIS11 family members identified so far in T84 and HT-29 human colorectal cancer cell lines. Butyrate response factor 1 (BRF1) transcription is repressed, butyrate response factor 2 (BRF2) transcription is activated and there is no apparent effect on the transcription of human TIS11 (HTIS11). Induction and repression occur rapidly, with altered mRNA levels detectable within 15 min of butyrate addition. Two other short-chain fatty acids, propionate and acetate, have no detectable effects on BRF1 or BRF2 transcription. PMID- 10367404 TI - Oxidative and malondialdehyde modification of low-density lipoprotein: a comparative study of binding and degradation by macrophages and endothelial cells. AB - Comparative study of oxidatively modified low-density lipoprotein (Ox-LDL) and malondialdehyde-modified low-density lipoprotein (MDA-LDL) is important for further understanding the biological properties of Ox-LDL, such as its toxic effects, immunogenicity and multiplicity of scavenger receptor binding. In this study, the characteristics of Ox-LDL and MDA-LDL binding and degradation were compared. The results show that when their degree of modification (as determined by relative electrophoretic mobility) was similar, the binding and degradation of Ox-LDL by the macrophage cell line P388D1 were greater than those of MDA-LDL. The binding and degradation of Ox-LDL by macrophages and human umbilical vein endothelial cells increased with the degree of modification. In addition, Ox-LDL or MDA-LDL could competitively inhibit binding of labelled Ox-LDL or labelled MDA LDL to their respective macrophage receptors, and could partially inhibit each other. PMID- 10367405 TI - In situ detection of superoxide anions within porcine articular cartilage. AB - Cartilage was isolated from pig articular joints, and the production of reactive oxygen species (ROS) by chondrocytes embedded within the cartilage was assessed by two methods: the reduction of nitro blue tetrazolium and by the use of diaminobenzidine in the presence of manganese ions. Little constitutive generation of ROS was seen, but it could be detected after the addition of the calcium ionophore ionomycin. Further, the response seen was extremely heterogeneous; some cells showed a far greater release of ROS than others. Cells arranged in the columnar arrays of the deep zone were the most active, while those furthest from the cartilage/bone interface (i.e. nearer to the outer face of the cartilage) were unresponsive. Chondrocytes cultured in alginate beads also showed a similar heterogeneity in their response, suggesting that the isolation of these cells and the measurement of ROS production in a population is not representative of the true situation. PMID- 10367406 TI - Apoptosis in developing rat teeth: a preliminary report. PMID- 10367407 TI - Evaluation of a screening medium for methicillin-resistant Staphylococcus aureus. PMID- 10367408 TI - Quantification of metabolic activity in tumour fragments. PMID- 10367409 TI - Rapid and simple Microtek biotyping method for Haemophilus influenzae. PMID- 10367410 TI - Angiogenesis: a new prognostic marker for breast cancer. AB - The past two decades have seen a huge increase in the study of angiogenesis or neovascularisation. With it has come realisation of the crucial role that angiogenesis plays in the growth and metastatic potential of carcinoma. One area where there has been particular interest is in the measurement of angiogenesis in breast carcinoma, with the hope that it may prove to be an independent prognostic indicator. As advances in breast cancer treatment have increased, so has the importance of assigning a patient to the optimal treatment regime. A number of studies have assessed tumour vascularity on sections immunostained with endothelial markers. Of these, the majority have found that highly vascular carcinomas have a poorer prognosis than those with low vascularity, although several studies have reported no correlation. These discrepancies may, in large part, be due to variations in the methods employed by different laboratories. At present, microvessel quantification in breast carcinoma has yet to prove it can reliably add a measurable amount of information to that already available from established prognostic indicators such as tumour size, nodal status, histological grade, and hormone receptor status. Although promising, further study is needed to establish the true value of measuring angiogenesis in breast cancer. PMID- 10367411 TI - The role of telomeres in ageing and cancer. AB - Telomeres are regions of DNA that cap the ends of linear chromosomes. In somatic cells the telomeres shorten progressively with every cell division, reducing the number of tandem repeat sequences. Eventually the chromosomes become unstable and the cell is no longer able to replicate. This represents an inherent biological clock in which the somatic cell has only a finite capacity for division. In contrast, germ cells do not undergo telomeric shortening and have relatively unlimited capacities for cell division. The difference is that germ cells retain the enzyme telomerase which is able to restore the telomere ends that are lost during cell division. Although telomerase activity is absent in most somatic cells, cancer cells acquire the ability to activate the enzyme, ensuring their immortal growth characteristics and selective advantage over normal somatic cells. PMID- 10367412 TI - Non-capsulate Haemophilus influenzae meningitis mimicking meningococcal disease. PMID- 10367413 TI - Issues underlying the evaluation of screening programmes. AB - Screening programmes have the potential to prevent premature death and disability and to improve quality of life, but they also have the potential for harm. Identifying worthwhile screening programmes, developing the correct strategies, and implementing them effectively is no easy task. A much more critical approach to screening is now being adopted and efforts are being made to ensure that new programmes of proven benefit and which are acceptable to the public, are effectively and equitably implemented in the community. We hope that this issue will stimulate further discussion and debate. PMID- 10367414 TI - Communication and interpretation of risk. AB - The 'new genetics' enables people's risk status for many diseases and disorders to be assessed much more accurately than before, yet considerable uncertainty remains over how risk information will be evaluated and acted upon. This paper summarises some of the main themes of psychological research on risk. Risk is traditionally defined in terms of probability. However, people often have difficulty in processing statistical information and may rely instead on simplified decision rules. Decision making under risk is also critically affected by people's subjective assessments of benefits and costs. In the field of genetic risk, such assessments may vary greatly between individuals, reflecting personal and cultural preferences and ethical concerns. The goals of risk communication should, therefore, not be merely the imparting of statistical 'facts' or the reduction of anxiety, but also enabling individuals and their families to make important decisions under conditions of uncertainty. PMID- 10367415 TI - Screening for prostate cancer: the current position. AB - Prostate cancer is a significant and increasing health problem in the UK and elsewhere, and there is considerable interest in the potential for screening. Of the currently available screening tests, measurement of serum levels of prostate specific antigen appears the most promising. However, despite evidence that screening can detect asymptomatic early stage disease, there is, as yet, no evidence that mortality from prostate cancer can be reduced. There are concerns that screening may result in considerable over-diagnosis of non-progressive or slowly developing disease, and the effectiveness of radical treatment of localised disease, which itself will cause some morbidity, remains a subject of debate. Population screening should not currently be recommended. Randomised controlled trials are in progress to assess the effectiveness of screening, but these will take many years to produce results. PMID- 10367416 TI - Should we be screening for colorectal cancer? AB - Mass population screening for asymptomatic neoplastic disease is now national policy in the UK for breast cancer and has been established for many years in the early diagnosis of carcinoma of the cervix. Cancer screening is based on the concept that treatment is more effective when the disease is localised and aims to detect it when it is at a less advanced clinico-pathological stage prior to the development of symptoms. Because colorectal cancer develops in benign adenomatous polyps which are often amenable to endoscopic resection, screening may both reduce the incidence of the disease as well as improving outcome from it. Flexible sigmoidoscopy screening focuses mainly on the detection of potentially malignant adenomas, their endoscopic removal producing a decrease in colorectal cancer incidence. It is a promising approach but conclusive data on effectiveness from a Medical Research Council-sponsored multicentre randomised controlled trial will not be available before 2006. Faecal occult blood testing aims to preferentially detect early stage invasive disease. Three randomised controlled trials of faecal occult blood screening show that the disease can be detected earlier in its development leading to reduced mortality from the disease -and that this is achieved at reasonable cost. The Department of Health is currently giving consideration to its national implementation. PMID- 10367417 TI - Screening for breast and ovarian cancer: the relevance of family history. AB - The recent identification of two breast and ovarian cancer susceptibility genes- BRCA1 and BRCA2--has received a lot of publicity. Public and professional expectations of the availability and utility of genetic testing have been raised and the importance of a family history of breast cancer overemphasized. In this chapter, we examine the significance of a family history of breast or ovarian cancer in determining individual risk. A strategy for management is proposed, based on stratifying women with such a history into three different categories of risk for breast cancer: high, moderate and low. Some of the more controversial aspects of screening for breast and ovarian cancer are reviewed, including the issue of management of women who are at increased risk of these cancers by virtue of a family history, genetic predisposition, or both. There is a need for further research to clarify the most appropriate management of those at moderate risk of developing these cancers. A management strategy for women at high risk is proposed. We believe that adoption of this strategy will strengthen consistent information giving from primary to tertiary care. PMID- 10367418 TI - Prenatal screening for chromosome abnormalities. AB - An abnormal chromosome complement (aneuploidy) contributes significantly to fetal loss during pregnancy, as well as to perinatal morbidity and mortality. The contribution of chromosomal abnormalities to fetal loss decreases as pregnancy continues with an estimated 50% of first trimester spontaneous abortions due to chromosomal abnormalities, but only 5% of stillbirths (after 28 weeks). Prenatal screening for aneuploidy (in particular Down syndrome) can be undertaken using maternal serum biochemistry, fetal ultrasound or a combination of both. In this chapter the advantages and disadvantages of screening programmes currently in use, or undergoing evaluation, will be reviewed. PMID- 10367419 TI - Screening for cystic fibrosis and its evaluation. AB - Cystic fibrosis (CF) is a recessively inherited disorder for which screening has been proposed. A number of different screening strategies have been suggested, including prenatal, preconceptional, school and neonatal carrier screening, as well as screening of newborns to identify affected infants. We discuss the advantages and disadvantages of these strategies, and identify gaps in knowledge relevant to decisions to introduce a screening programme for cystic fibrosis. Screening to identify carriers during the newborn period or among school age children is inadvisable, mainly on psychosocial and cost-effectiveness grounds. Although early diagnosis of CF may improve prognosis, current scientific evidence is not sufficient to support screening newborns to identify affected infants. of the remaining two options, prenatal screening has a practical advantage because of existing facilities, while with screening before conception all reproductive options are, in principle, open to detected carrier couples. If adequate pre- and post-test counselling can be provided, both two types of screening could be introduced. PMID- 10367421 TI - Screening for genital chlamydial infection. AB - Genital chlamydial infection is a common, sexually transmitted infection that is often asymptomatic, but associated with long term morbidity in a sizeable proportion of women. Early infection can be diagnosed reliably using noninvasive methods and treated effectively with antibiotics. The case for screening in conventional high risk settings (e.g. genito-urinary medicine and termination of pregnancy clinics) is already clear. Screening in the wider community also needs evaluating if a significant impact on the problem is to be made since chlamydial infection is widely distributed among young, sexually active people who may have little contact with health services. Studies are in progress to assess the acceptability of different screening approaches to women and men in the community and to compare performance of newer diagnostic techniques. The cost-effectiveness of community-based screening in reducing morbidity needs to be evaluated empirically in randomised trials to encourage a coherent, evidence-based screening policy in this country. PMID- 10367420 TI - Evaluating newborn screening programmes based on dried blood spots: future challenges. AB - A UK national programme to screen all newborn infants for phenylketonuria was introduced in 1969, followed in 1981 by a similar programme for congenital hypothyroidism. Decisions to start these national programmes were informed by evidence from observational studies rather than randomised controlled trials. Subsequently, outcome for affected children has been assessed through national disease registers, from which inferences about the effectiveness of screening have been made. Both programmes are based on a single blood specimen, collected from each infant at the end of the first week of life, and stored as dried spots on a filter paper or 'Guthrie' card. This infrastructure has made it relatively easy for routine screening for other conditions to be introduced at a district or regional level, resulting in inconsistent policies and inequitable access to effective screening services. This variation in screening practices reflects uncertainty and the lack of a national framework to guide the introduction and evaluation of new screening initiatives, rather than geographical variations in disease prevalence or severity. More recently, developments in tandem mass spectrometry have made it technically possible to screen for several inborn errors of metabolism in a single analytical step. However, for each of these conditions, evidence is required that the benefits of screening outweigh the harms. How should that evidence be obtained? Ideally policy decisions about new screening initiatives should be informed by evidence from randomised controlled trials but for most of the conditions for which newborn screening is proposed, large trials would be needed. Prioritising which conditions should be formally evaluated, and developing a framework to support their evaluation, poses an important challenge to the public health, clinical and scientific community. In this chapter, issues underlying the evaluation of newborn screening programmes will be discussed in relation to medium chain acyl CoA dehydrogenase deficiency, a recessively inherited disorder of fatty acid oxidation. PMID- 10367422 TI - Screening for abdominal aortic aneurysms. AB - Ruptured aneurysm of the abdominal aorta is a common preventable cause of death, accounting for 2% of all deaths in men over 60 years of age. Population screening could prevent such deaths. Aortic diameter (which can be measured accurately on ultrasound) is a strong predictor of the risk of rupture, which is about 17% per year with aortic diameter > or = 6 cm, but below 0.5% per year with aortic diameter < 5 cm, with uncertainty regarding risk in the range 5.0-5.9 cm. Adopting an aortic diameter cut-off of 6.0 cm, the detection rate is estimated to be 86% (that is, 86% of all men who would rupture an aortic aneurysm could be identified and offered surgery) and the false positive rate only 0.6% (that is, 0.6% of men who would not rupture an aortic aneurysm would be so identified). In men with aortic diameter > or = 6 cm, the risk of rupture of 17% per year greatly outweighs the peri-operative mortality of about 5%. A national screening programme for men over 60 years of age could prevent 2000 deaths per year and should commence. Uncertainty remains regarding the frequency with which men with smaller aneurysms should be re-examined and the value of intervention among those with an aortic diameter of 5.0-5.9 cm, but the screening programme itself would generate data to help resolve these issues. PMID- 10367423 TI - Is screening for osteoporosis worthwhile? AB - Osteoporosis is a common condition, which is recognised by the occurrence of fragility fractures and leads to considerable mortality and morbidity with huge financial implications world-wide. Based on predicted demographic changes, the implications of this disease are set to increasingly affect the healthcare budgets of all nations. The determinants of fracture are skeletal factors, such as peak bone mass, the rate of bone loss and extra-skeletal factors, which include trauma and the response to that trauma. Some of these factors are genetically determined, but several have environmental origins, which could, theoretically, be manipulated. There are two potential means whereby osteoporotic fractures might be prevented. Measures could be targeted at the entire population, with the aim of shifting the distribution of bone mass in a beneficial direction, through modifying the behaviour of all individuals. The alternative is a high risk approach, whereby intervention is targeted only at those considered to have the greatest risk of future fracture. Mass bone density screening falls into the second approach. Bone density is a good predictor of future fracture risk, and cost-effectiveness analyses of the high risk approach suggest economic benefits of policies targeting pharmacological treatment to those individuals at highest risk. However, there are important concerns about the levels of compliance achievable with such therapeutic interventions, the balance of risks and benefits for some of these interventions (for example, hormone replacement therapy), and the outcome when treatment is discontinued. On current evidence, it is certainly not appropriate to target hormone replacement therapy for women at the menopause on the basis of a bone density screening programme. However, newer bone-specific agents are being developed which might be administered at later ages, closer to the time when fracture incidence rates rise steeply. Bone densitometry has been shown to predict fractures even in the elderly, and high risk strategies for the targeting of such agents (for example, the bisphosphonates or selective oestrogen receptor modulators) will remain important research issues for the future. PMID- 10367424 TI - Screening in child health. AB - Screening programmes in child health have evolved on the basis of individual enthusiasm and professional consensus, rather than being based on objective evidence of benefit. Three reviews have been carried out in the UK over the past 10 years. The only programmes which show robust evidence of effectiveness are those for PKU and hypothyroidism. The value of screening for hearing loss and vision defects is widely accepted, but there are many unresolved issues. Programmes for detection of congenital dislocation of the hip, congenital heart disease and growth disturbances are of doubtful value. Early identification of developmental problems is stressed by parents, but screening may not be the best way to achieve this. The UK programme of well-child care places increasing emphasis on promotion of physical and emotional health; screening tests should either be subjected to quality monitoring, or removed from the programme if they cannot fulfil the classic criteria of Wilson and Jungner. PMID- 10367425 TI - Multidimensional assessment of elderly people in the community. AB - Multidimensional assessment has been advocated as the most appropriate type of screening activity for elderly people. The emphasis has traditionally been on screening for problems with function, disability and dependency in order to identify service and treatment needs. Several randomised trials of multidimensional assessment have been conducted. The UK trials have been conducted in the setting of general practice, while trials in other European countries as well as the US have targeted elderly people living in the community. Although there appear to be possible benefits from multidimensional assessment, for example in reduced mortality, disability and hospital inpatient admissions, these trials have not been consistent in their findings, nor have they been large enough to produce results of sufficient precision and certainty to inform policy. There is stronger evidence that multidimensional assessment can prevent falls but the size of the benefit for serious falls is quite small. The UK health policy of regular assessment of people aged 75 years and above to be carried out in general practice has been implemented haphazardly with little guidance on appropriate methods and levels of assessment. A large randomised trial is currently underway in the UK which will provide evidence on the cost effectiveness of a range of different strategies for assessment. PMID- 10367426 TI - Screening in general practice and primary care. AB - General practice and its associated primary care services are the final common pathway for the delivery of most screening programmes. The absence of nationally agreed priorities, guidelines and identifiable resources has meant that screening in primary care remains somewhat arbitrary, practice varies widely and programmes remain largely unevaluated. Discussion of screening has focused largely on test characteristics and performance with less attention being given to issues of policy formation, priority setting, implementation and quality assurance. Without these elements, quality and test performance deteriorate, recruitment and follow up are incomplete and a poorly discriminating test of doubtful utility is applied inequitably and inefficiently. For general practice there are two major concerns. The first is to improve delivery of programmes of proven efficacy, such as breast or cervical screening, that already have a national framework. The second is to develop and provide a national structure for preventive programmes for cardiovascular and smoking-related disease. For cardiovascular disease, the issue is no longer whether to screen and advise whole populations for multiple risk factors, but how best to implement this programme. In this chapter, the case for screening for cardiovascular disease is reviewed and potential strategies for improving delivery of screening in general practice and primary care discussed. PMID- 10367427 TI - Quality assurance in screening programmes. AB - All screening programmes do harm; some also do good. The responsibility of the policy-maker is to decide which programmes do more good than harm at reasonable cost and then introduce them, once they are confident that the screening programme could and will reach the standard of quality required for success. The ratio of benefit to harm is not, however, constant and this relationship demonstrates a shifting balance. PMID- 10367428 TI - The economic perspective. AB - The rationale for the economic perspective on screening is presented and the particular relevance of economic evaluation highlighted. The principles of economic evaluation are described in terms of measuring and valuing the costs and outcomes associated with screening. The different types of economic evaluation are described with discussion of data sources. The problematic issues associated with time preferences and discounting and with the measurement and valuation of outcomes other than true positives are discussed. Issues associated with antenatal screening, in particular the inclusion of averted costs due to the termination of an affected pregnancy and the inclusion and valuation of the unborn child's utility, are also raised. PMID- 10367429 TI - Ethical and legal issues. AB - This article considers the general issues surrounding screening, in particular the problems involved in the collection and use of information derived from screening. Different forms of screening entail particular problems, which have their own ethical complexities. The legal and ethical issues arising from screening are likely to assume greater significance. The screening opportunities afforded by the Human Genome Project will provide humanity with choices that would have been inconceivable to any previous generation. PMID- 10367430 TI - Making media messages more suitable for health education in the African context. AB - The design of appropriate health education messages and materials is not an easy task, because a complex range of interrelated variables determine the level of message appropriateness. This article focuses on the socio-cultural dimension of health education message design. Messages that are perceived as culturally inappropriate could lose credibility among certain audiences and could even lead to polarization of health beliefs and reinforce high risk behaviour in some communities. Apart from theoretical viewpoints and principles, the article presents examples of appropriate visuals and text within the African context. Practical guidelines are offered to improve the appropriateness of media messages for health education in Africa. PMID- 10367431 TI - Photo CD for clinical imaging. AB - This paper examines the use of Photo CD as a source for printing clinical images for patients' medical case notes. Advantages include multiple resolutions, no capital investment in digital equipment, and professional and flexible presentation of pictures in patients' notes. Disadvantages include increased running costs for every film, a limited number of images stored on each CD and the requirement to set up a comprehensive image database to store and retrieve digital images. PMID- 10367432 TI - A clinical image library using photo CD. AB - In a move to update a collection of medical slide images and improve access to high-quality medical images for postgraduate teachers at the Institute of Child Health and Great Ormond Street Hospital for Children, a new collection of scanned slide images was created on Kodak Photo CD discs. Each image was available in five different resolutions. The images were catalogued in an image database by diagnosis, date, patient registration number, and anatomical view. Images could be accessed using simple search protocols and output to digital files, slides or prints. PMID- 10367433 TI - Slide presentations. AB - Although my discomfiture is directed at the lecturer, my criticism is of the slides used. I would like every lecturer and illustrator to vow never to use one slide if five will do better, and never to use a slide that needs a pointer. PMID- 10367435 TI - Estimation of evoked potentials using total least squares prony technique. AB - The authors investigate the applicability of Prony modelling to the estimation of evoked potentials. Four types of total least squares (TLS) model are considered and their optimal parameters are defined based on ten visual averaged EPs. Simulations with various signal and noise characteristics show that the TLS-Prony estimation is superior to averaging for two of the models, namely the unconstrained and the stable models. Application of the TLS-Prony estimator as a post-processor to moderate averaging allows a reduction in the number of responses averaged, or equivalently of recording time, by a factor of two. PMID- 10367436 TI - Prediction of epileptic seizures from two-channel EEG. AB - Multivariate spectral estimation based on parametric modelling has been applied to epileptic surface EEG in order to detect EEG changes that occur prior to the clinical outbreak of the seizure. A better time/frequency resolution has been achieved using residual energy ratios (Dickinson's method). Prediction of oncoming seizures was based on detection of increased preictal synchronisation by calculation of coherence and pole trajectories. The method has been tested on simulated EEG data and on real EEG data from patients with primary generalised epilepsy. Prediction times of 1-6 s have been found in several seizures from five patients. PMID- 10367434 TI - Developments in cardiovascular ultrasound. Part 3: Cardiac applications. AB - Echocardiography is still the principal, non-invasive method of investigation for the evaluation of cardiac disorders. Using Doppler ultrasound, indices such as coronary flow reserve and cardiac output can be determined. The severity of valvular stenosis can be determined by the area of the valve, either directly from 2D echo, from pressure half-time calculations, from continuity equations or from the proximal isovelocity surface area method. Alternatively, the severity of regurgitation can be estimated by colour or pulsed ultrasound detection of the back-projection of the high-velocity jet into the chamber. Myocardial wall abnormalities can be assessed using 2D ultrasound, M-mode or analysis from the radio-frequency-ultrasound signal. Doppler tissue imaging can be used to quantify intra-myocardial wall velocities, and 3D reconstruction of cardiac images can provide visualisation of the complete cardiac anatomy from any orientation. The development of myocardial contrast agents and associated imaging techniques to enhance visualisation of these agents within the myocardium has aided qualitative assessment of myocardial perfusion abnormalities. However, quantitative myocardial perfusion has still to be realised. PMID- 10367438 TI - Evaluation of catheter-mounted transducers for intra-oesophageal pressure recording in respiratory function tests. AB - Oesophageal pressure measurements in respiratory function tests are commonly performed using a balloon-catheter system. This study investigates the usefulness of catheter-mounted pressure transducers as an alternative to balloon-catheter systems. Calibration related physical properties of the catheter mounted pressure transducers are evaluated in vitro. The behaviour of these transducers in vivo is evaluated in ten volunteers by relating pressures measured in the oesophagus to airway opening pressures and by comparing these relationships with those sequentially obtained by a balloon-catheter system. The catheter-mounted pressure transducers show no drift after a proper preparation procedure. These catheters, with integrated pressure transducers, are tolerated significantly better by the subjects than are balloon catheters. The catheter-mounted pressure transducers are found to give an equivalent performance compared with the balloon-catheter system, if relative pressures are of interest. However, unpredictable and uncontrollable shifts in offset occur during the in vivo measurements, disturbing absolute pressure readings. Possible explanations for these shifts are the presence of bubbles and adhesion of mucus to the transducers, exerting Van der Waals forces, and contact with the tissue of the oesophageal wall. These shifts are found to be quite stable throughout a period of measurement and therefore of minor disturbance to relative pressure measurements, for instance in assessing the elastic properties of lungs. PMID- 10367437 TI - Technical aspects of mechnomyography recording with piezoelectric contact sensor. AB - The piezoelectric contact sensor has been widely utilised in mechanomyography (MMG). The authors aim to clarify the mechanical variables (i.e. acceleration, velocity or displacement) reflected by the MMG signal detected with a piezoelectric contact sensor (PEC), and compare the results with those obtained simultaneously by an accelerometer (ACC). To measure the acceleration-frequency response, a mechanical sinusoidal excitation of 5 to 300 Hz at a constant magnitude of 0.01 G was applied to the two transducers. The acceleration frequency response of the ACC transducer was confirmed to be almost flat. The PEC without any restriction of the transducer housing (including the combined seismic mass) demonstrated a similar response to the ACC transducer. The PEC transducer output with restricted housing decreased with increasing sinusoidal frequency and an attenuation slope of -40 dB/decade and phase angle of -180 degrees. The voluntary MMG signal during isometric knee extension was recorded simultaneously with the two transducers. The amplitude spectral density distribution of the MMG from the PEC transducer was narrow and the mean frequency was approximately one half that obtained from the ACC transducer. The amplitude spectral density distribution with the PEC transducer resembled that of the double integral over time of the ACC transducer signal. The phase angle of the PEC transducer signal was different from that of the ACC transducer signal by approximately -180 degrees. These results suggest that the PEC transducer acts as a displacement meter of muscle vibration. In addition, differences in the MMG frequency components relating to the transducer type must be taken into consideration when investigating the mechanical activity of muscle. PMID- 10367439 TI - Wavelet based ST-segment analysis. AB - A novel algorithm for ST-segment analysis is developed using the multi-resolution wavelet approach. The system detects the QRS complexes and analyses each beat using the wavelet transform to identify the characteristic points (fiducial points). These fiducial points are, iso-electric level, the J point, and onsets and offsets of the QRS complex and T wave. The algorithm determines the T onset by looking for a point of inflection between the J point and the T peak. Furthermore, detection of characteristic points by the wavelet technique reduces the effect of noise. The results show that the proposed approach gives very accurate ST levels, as compared to the conventional (empirical) technique, at higher heart rates and with different morphologies. The algorithm detects the ST segment length in 92.3% beats with an error of 4 ms, and in 97.3% beats the error is within 8 ms. The algorithm has been implemented on a TMS320C25 based add-on DSP card connected to a PC to provide the on-line analysis and display of ST segment data. PMID- 10367440 TI - Comparison of heart rate variability spectra using generic relationships of their input signals. AB - Spectral analysis of heart rate variability (HRV) is an accepted method for assessment of cardiac autonomic function and its relationship to numerous disorders and diseases. Various non-parametric methods for HRV estimation have been developed. The spectrum of counts, the instantaneous heart rate spectrum and the interval spectrum are mostly practised. Although extensive literature on their respective properties is available, there seems to be a need for a more complete comparison, eventually resulting in recommendations for applicability. The methods for HRV spectral analysis use their specific transforms of the primary R-R interval series into input signals for spectral computation. This is, in fact, the reason for obtaining different spectra. A basis for comparison is established, revealing the generic relationships of these HRV input signals. It allows for a more systematic evaluation and for further development of the considered methods. The results with simulated and real HRV data show better performance by the spectrum of counts and by a proposed instantaneous heart rate spectrum, obtained using a cubic spline interpolated input signal. The modulation depth of the primary signal can influence the accuracy of the HRV analysis methods. PMID- 10367441 TI - Application of a wavelet adaptive filter to minimise distortion of the ST segment. AB - A wavelet adaptive filter (WAF) for the removal of baseline wandering in ECG signals is described. The WAF consists of two parts. The first part is a wavelet transform that decomposes the ECG signal into seven frequency bands using Vaidyanathan-Hoang wavelets. The second part is an adaptive filter that uses the signal of the seventh lowest-frequency band among the wavelet transformed signals as primary input and a constant as reference input. To evaluate the performance of the WAF, two baseline wandering elimination filters are used, a commercial standard filter with a cutoff frequency of 0.5 Hz and a general adaptive filter. The MIT/BIH database and the European ST-T database are used for the evaluation. The WAF performs better in the average power of eliminated noise than the standard filter and adaptive filter. Furthermore, it shows a lower ST-segment distortion than the standard filter and the adaptive filter. PMID- 10367442 TI - Implication of the systolic hump in systemic arterial pressure waves. AB - The systolic hump in the aortic blood pressure wave is defined as the aortic resistance component proportional to the aortic blood flow superimposed on the windkessel component. An electrical analogue comprising a series resistance (aortic resistance) plus a resistance (peripheral resistance) and capacitance (aortic compliance) in parallel (i.e. windkessel component) is used for analysis. Curve fitting using the least-squares method is performed on calculated and measured blood pressure waves from dogs under haemodynamical conditions induced by infusion of three drugs (noradrenaline, isoproterenol and acetylcholine). The curve fitting RMS (root mean square) errors are < 3% for blood pressure waves and < 30% for blood flow waves, with good agreement between measured and calculated blood flow waveforms. Infusion of noradrenaline and acetylcholine is found to induce a significant decrease and increase in the aortic resistance, respectively. Although only a small fraction of the blood pressure wave, the systolic hump has a marked effect on the systolic pressure waveform. PMID- 10367443 TI - Thoracic geometry and its relation to electrical current distribution: consequences for electrode placement in electrical impedance cardiography. AB - In thoracic impedance cardiography (TIC) measurements the neck electrodes are often positioned at the basis of the neck, close to the neck-thorax transition. Theoretically, this neck-thorax transition will cause inhomogeneities in the current density and potential distribution. This was simulated using a 3D finite element method, solely representing the geometrical neck-thorax transition. The specific conductivity was 7 10(-3) (omega cm)-1 and the injected current was 1 mA. As expected, the model generated inhomogeneities in the current distribution at the neck-thorax transition, which reached as far as 5 cm into the neck and 20 cm into the thorax. These results are supported by in vivo measurements performed in 10 young male subjects, in which the position of the neck electrodes was varied. A two-way ANOVA revealed that the stroke volume of the lowest neck position was significantly different from the other positions. Small shifts in the position of the neck electrode resulted in large changes in impedance and stroke volume (127 to 82 ml for the Kubicek equation). To standardise the electrode position, the authors strongly recommend placement of the neck electrodes at least 6 cm above the clavicula. PMID- 10367444 TI - An electrically isolated balanced wideband current source: basic considerations and design. AB - At relatively high frequencies, the application of an alternating current through the body or a body segment results in electromagnetic stray fields which reduce the amount of current actually injected into the tissue under study. This radiation effect can be reduced by use of a symmetrical configuration current source. The symmetry of such an arrangement, however, depends on the stray capacitances of the source with respect to surrounding equipment. To minimise these effects, it is required that the source is electrically isolated from the surrounding equipment and the subject under study. In this manner stray capacitances with respect to elements of the current source are reduced. In such a configuration common mode voltages to the input amplifier of the measuring system are also reduced. The paper describes design considerations and the implementation of a wideband current source capable of injecting alternating current in the order of 300 microARMS into biological tissue having impedances up to 1 k omega. Current stabilisation is obtained by means of a control circuit which measures the actual current passing through the tissue under study. Leakage currents arising from shielding and stray capacitances are compensated for. The usable frequency range is between 4 kHz and 1024 kHz and current stability is better than 0.2%. Through the use of a symmetrical, floating circuit a configuration is obtained which substantially reduces stray effects. The current source is connected to other circuits by means of two isolation ports: (1) a transformer coupling for the carrier frequency; and (2) an opto-coupler to transfer a phase reference signal obtained from current measurement. The current amplitude can be modulated by controlling the reference input to the control loop by means of a third auxiliary isolation port for transfer of the modulating signal. PMID- 10367445 TI - Impedance of goat eye lens at different DC voltages. AB - A computer assisted AC impedance system is used to measure the DC voltage-current (V-I) characteristics and AC impedance of a goat eye lens using a two-probe Ag AgCl electrode system. The measurement of the V-I characteristics shows that when a DC voltage from 0 mV to 30 mV is applied, the resultant current decreases from an initial value of 0.58 microA to 0.006 microA. However, when the voltage is increases beyond 30 mV, the current increases and reaches a value of 0.9 microA at 100 mV. The data on the frequency response (0.01-10 Hz) of the impedance of lens tissue show an inverse relationship with frequency. The effect of various DC voltages, namely 0, 30, 50, 100 and 200 mV, on the impedance of the eye lens is also investigated over a frequency range of 0.01-10 Hz. The measurement results for both V-I characteristics and AC impedance further suggest the presence of a 30 mV voltage compartment in the goat eye lens. PMID- 10367446 TI - Feature extraction and state identification in biomedical signals using hierarchical fuzzy clustering. AB - Many problems in the field of biomedical signal processing can be reduced to a task of state recognition and event prediction. Examples can be found in tachycardia detection from ECG signals, epileptic seizure or psychotic attack prediction from an EEG signal, and prediction of vehicle drivers falling asleep from both signals. The problem generally treats a set of ordered measurements and asks for the recognition of some patterns of observed elements that will forecast an event or a transition between two different states of the biological system. It is proposed to apply clustering methods to grouping discontinuous related temporal patterns of a continuously sampled measurement. The vague switches from one stationary state to another are naturally treated by means of fuzzy clustering. In such cases, an adaptive selection of the number of clusters (the number of underlying semi-stationary processes) can overcome the general non stationary nature of biomedical signals and enable the formation of a warning cluster. The algorithm suggested for the clustering is a new recursive algorithm for hierarchical fuzzy partitioning. Each pattern can have a non-zero membership in more than one data subset in the hierarchy. A 'natural' and feasible solution to the cluster validity problem is suggested by combining hierarchical and fuzzy concepts. The algorithm is shown to be effective for a variety of data sets with a wide dynamic range of both covariance matrices and number of members in each class. The new method is applied to state recognition during recovery from exercise using the heart rate signal and to the forecasting of generalised epileptic seizures from the EEG signal. PMID- 10367447 TI - Dust anchoring characteristics of electret fibres with respect to Der p 1 allergen carrying particles. AB - The avoidance of house dust mite allergens is a major area of interest and essentially requires a significant removal of these allergens from the immediately respirable air. Electrostatic attraction and anchoring of particulate matter using electret polymers is commonly used for air filtration purposes. This effect is investigated for its possible use in domestic allergen avoidance. Polypropylene electret, heat-treated electret and non-electret, and wool and nylon fibre samples were soiled with house dust known to contain Der p 1 allergen. These samples were vacuumed at three air face velocities. The proportions of released and anchored dust were calculated. Released dust was collected and analysed for Der p 1 concentration and compared to stock dust values. Results showed that compared to uncharged fibres at least 95% more dust remained anchored in the electret fibres. Also, overall Der p 1 release was reduced by more than 49%. Der p 1 allergen concentrations in the collected dust were relatively constant for all the fibres tested, indicating no selective attraction or repulsion of Der p 1 allergen carrying particles in the experimental dust. The consistently high dust anchoring ability of the electret fibres could be used in many domestic products that are known to harbour particulate allergens, to reduce their release and inhalation. PMID- 10367448 TI - Quantification of the morphological features of a full microvascular network. AB - Investigations into the changes that occur in microvasculature following the surgical procedure called delay have brought about the need for a computer system capable of quantifying the morphological features of a full microvascular network in terms of average vessel length, diameter, and tortuosity. Both the formulaic conventions that have been developed to measure these quantities as well as their implementation in the form of a HP-9000/UNIX based computer software system that we developed specifically for this purpose are discussed. Reliability studies performed using the final system to measure the microcirculatory network of a mouse latissmus dorsi muscle (LDM) showed 95% confidence intervals within 5% of means and coefficients of variability within 7% of means for all quantities measured in large (150-300 microns), medium (50-150 microns), and small (< 50 microns) diameter vessels. These variations were significantly smaller than the changes that were observed in a preliminary study comparing these microvascular network parameters before and after delay in the hairless mouse LDM, showing the proposed quantification methods to be well suited to the study of the microvascular changes following delay. It is hoped that the formulaic conventions, implementation process and reliability data will provide a useful comparison for other researchers interested in measuring similar features of microcirculatory networks. PMID- 10367449 TI - High-speed electromechanical shutter for vision research. AB - A low-cost and high-speed electromechanical shutter is described. The design makes use of a small magnet moving in a magnetic field and includes a specially developed electronic control for optimal operation. The opening and closing time of the shutter is below 200 microseconds with overall time for generating light flashes close to 1 ms. PMID- 10367450 TI - Measurement of sealing resistance of cell-electrode interfaces in neuronal cultures using impedance spectroscopy. AB - Sealing resistance is highly significant with respect to the electrical neuron electrode contact because it decreases the stimulation threshold of neurons cultured on a planar micro-electrode array. A method is proposed for measurement of the sealing resistance using impedance spectroscopy. The effect of the sealing resistance on the total impedance spectrum of a cell-electrode interface is modelled for complete coverage of the electrode by the cell. Sensitivity analysis demonstrates that the impedance spectrum is determined by four parameters: two electrode parameters, the sealing resistance and the shunt capacitance between the lead of the electrode and the culture medium. Experimental verification of the model is performed by simultaneous measurement of the impedance spectrum and electrode coverage. A good and unique fit between the simulated and measured impedance spectra was obtained by varying the two electrode parameters and the sealing resistance. PMID- 10367451 TI - Preliminary study on the suitability of a pharmacological bio-assay based on cardiac myocytes cultured over microfabricated microelectrode arrays. AB - There are a range of techniques that can be used to assay bioactive compound. One potentially promising technique is a system consisting of microfabricated extracellular recording devices over which electrogenic cells can be grown. To date, research in this area has concentrated on the use of neurons as an electrogenic cell type. However, these cells have limitations. Only small extracellular potentials have been recorded from mammalian neurons cultured over microfabricated electrode arrays. Although such potentials may be of use in assays examining the effects of bio-active compound analogues on firing frequency, they are of little use for more detailed pharmacological studies involving analyses of signal shape. What is required is a system from which much larger extracellular potentials can be recorded. This preliminary study reports on a system based on cardiac myocytes cultured over microfabricated metal microelectrode arrays, from which potentials with a mean amplitude of 16.9 microV can be reliably recorded, which can be reversibly blocked with mumoll-1 concentrations of the sodium ion channel blocker lidocaine. Less common potentials with amplitudes of up to 3.5 mV were also recorded. It is demonstrated that cardiac myocytes cultured over microfabricated micro-electrode arrays can be used in assays of cardioactive compound analogues. PMID- 10367452 TI - Liposome-induced release of cell membrane proteins from intact tissue epithelium. AB - During extraction and purification, membrane proteins very often undergo denaturation and deactivation. To overcome this problem, the authors have tried to establish a better methodology to make the study at in vivo tissue level, not at the isolated cellular level, possible and easier. This is in vivo direct exposure of animal tissue to the liposome that contains an artificial boundary lipid (D14DPC, 1,2-dimyristamido-1,2,-deoxyphosphatidylcholine). Bullfrog and rat tongues were used. To confirm the reasonableness of this methodology, several different techniques were adopted; the nerve response study, gel electrophoretic analysis, quartz crystal microbalance (QCM) measurement and the affinity gelchromatography. When the tongue was exposed to the D14DPC-containing DMPC liposome, a significant amount of membrane protein was found in the recovered liposome (this was the production of proteoliposome). The nerve response in the neurophysiological measurement to several taste stimuli, such as L-alanine, L leucine, sucrose and quinine hydrochloride significantly decreased when the tongue was exposed to the same liposome. These phenomena were common to both bullfrog and rat tongues. The nerve response to the stimulation with L-alanine was the most remarkably affected in the liposomal treatment. Therefore, the L alanine-binding protein was focused upon to confirm the reasonableness of the QCM measurement and the affinity gelchromatography. The D14DPC-containing proteoliposome always showed significant binding to both the L-alanine affinity gel and the L-alanine-conjugated QCM. The results revealed that membrane proteins can be directly and effectively released, even from intact animal tissue epithelium, using the artificial boundary lipid-containing liposome. PMID- 10367453 TI - Cell culture approach to biocompatibility evaluation of unconventionally prepared hydroxyapatite. AB - Hydroxyapatite (HA), Ca10(PO4)6(OH)2 was produced by microwave irradiation of calcium nitrate (CaNO3.4H2O) and di-ammonium phosphate in aqueous solution. The HA formation was confirmed by X-ray diffraction analysis. HA prepared by this unconventional route was subjected to biocompatibility assay by a cell-culture method using the hybridoma cell line AE9D6 in both conventional Dulbecco's modification of Eagle's medium (DMEM) and simulated body fluid (SBF), both supplemented with 5% fetal calf serum. HA synthesised through this unconventional method showed the presence of tricalcium phosphate which can be reduced only after heat treatment at 1150 degrees C. The HA conformed to the X-ray data index file for hydroxyapatite. Biocompatibility assays showed reproducible growth and secretion patterns of cells both in DMEM as well as in SBF, thereby indicating the effectiveness of this method for the production of biocompatible HA. PMID- 10367454 TI - Volumetric analysis of CT orbital images. AB - The determination of the volume of eye muscles is a problem of considerable interest in Graves' ophtalmopathy both with respect to the diagnostic quantisation of the disease and to the measured assessment of the effectiveness of pharmacological treatment. The aim of this research is to design and test an advanced method for processing computerised tomography (CT) orbital images in order to obtain a three-dimensional reconstruction of infra-orbital muscle structures and to analyse them from a morphometric viewpoint. CT images of subjects suffering from Graves' disease were acquired before and after pharmacological treatment with immunosuppressors. They were then processed along with CT images of an anatomical phantom of known volume and fusiform morphology in order to assess the reliability of the procedure and to calculate the effect of the different modalities of acquisition and processing of CT images on the error in volume calculation. PMID- 10367455 TI - Automated sizing of DNA fragments in atomic force microscope images. AB - Current techniques used to measure lengths of DNA fragments in atomic force microscope (AFM) images require a user to operate interactive software and execute tedious error-prone cursor selections. An algorithm is proposed which provides an automated method for determining DNA fragment lengths from AFM images without interaction from the computer operator (e.g. cursor selections or mouse clicks). The approach utilises image processing techniques tailored to characteristics of AFM images of DNA fragments. The automated measurements have a mean absolute deviation of less than 1 pixel when compared to manual image-based measurements. The DNA length determined from the histogram of calculated lengths is accurate to within 3% of the actual DNA length in solution. For fragments that are 250 base-pairs long, the precision is estimated to be within 17 nm, which is about 20% of the total length. This precision was confirmed when the algorithm easily resolved fragments in one image that differed by only 17 nm. Fragment sizes up to 2000 base-pairs have been tested and successfully sized. This algorithm is being developed as part of a new solid-state DNA sizing technique for applications such as genotyping and construction of physical genome maps. PMID- 10367456 TI - Non-invasive temperature imaging of muscles with magnetic resonance imaging using spin-echo sequences. AB - The application of spin-echo magnetic resonance imaging sequences on non-invasive temperature imaging for temperature mapping of human limbs is investigated. In an in vitro experiment performed on a meat sample, the equilibrium magnetisation P and the spin-lattice relaxation time T1 are calculated from the values for the repetition time TR and the signal intensities obtained by a spin-echo sequence at different tissue temperatures as measured by a fibre-optic probe. T1 is linearly correlated to the tissue temperature, and P is linearly correlated to the reciprocal value of the absolute temperature. Both effects, taken together, lead to a non-linear dependency of the signal intensity on temperature. Therefore a TR leading to maximum temperature dependency of the signal intensity is calculated and used in the further experiments. In the in vivo experiments, the lower legs of two volunteers are cooled from outside. Images are acquired with a spin-echo sequence (1.5 T, TR = 1200 ms, TE = 10 ms). A rise in signal intensity in the muscle with falling skin temperature is observed, particularly in more peripheral muscle layers. This study shows that spin-echo sequences can be used to monitor temperature changes and temperature differences in living muscle tissue. PMID- 10367457 TI - Roughness feature of metaphase chromosome spreads and nuclei for automated cell proliferation analysis. AB - As a step towards automation of mitotic index estimation for cell proliferation studies, a roughness feature of surface-intensity images is introduced: the mean depth-width ratio of extrema (MDWRE). This feature allows identification of variable-shaped metaphases and interphase nuclei in the presence of many artefacts (one metaphase per hundreds of nuclei and thousands of artefacts). The texture of the cytological objects (seen as rough surfaces) is quantified by scanning, in one dimension, the lines contained in a closed contour. MDWRE proves to be suitable for image magnifications by a factor of as low as ten, making faster scanning of slides possible. The use of this feature gives +14%, +65%, +133% and +133% better performance figures than classical textural features derived from co-occurrence matrices, such as contrast, energy, entropy and angular second moment, respectively, and +51% better than the relative extrema density (RED). The MDWRE per object and the shape of the histogram of the depth width ratio of grey-level roughs have been shown to be very useful as textural features for the classification of metaphase images. PMID- 10367458 TI - Three-dimensional biomechanical model for simulating the response of the human body to vibration stress. AB - Several investigations have revealed that long-term exposure to whole-body vibrations can induce low back pain. In analogy to materials handling, the health risk can be assessed if the forces transmitted in the spine during vibration are known. To estimate the forces a biomechanical model has been developed in which the human trunk, neck, head and arms are represented by 16 rigid bodies. An additional body simulates the vibrating seat. The bodies are connected by visco elastic joint elements, and 56 force elements imitate the trunk and neck muscles. The motion equations are derived by means of the dynamics of systems of rigid bodies, and the motions are simulated in three directions. The frequency-response functions between the accelerations of the seat and the head satisfactorily correspond to data reported in the literature. The spine forces are composed of a static part, due to body posture, and a vibration-induced part. The relation between the oscillating parts of the forces transmitted from seat to pelvis and the spine forces are also described by frequency-response functions. To assess the health risk the simulated spine forces must be compared with the strength of the spine, bearing in mind that this is dependent on the number of load cycles. PMID- 10367459 TI - Implementation and application of real-time motion analysis based on passive markers. AB - A method for real-time motion analysis based on passive markers is presented. An opto-electronic automatic motion analyser was used as hardware platform and the real-time operation was based on the interfacing between two levels of the system architecture. True real-time acquisition, processing and representation of two dimensional and three-dimensional kinematics data were implemented through a newly conceived data acquisition procedure and high speed optimisation of the kinematics data processing. The method allows one to operate the motion analysis system in real-time; even when the data elaboration unit is required to perform other processing functions, the only consequence is a decrease in system sampling rate. The maximum number of processed and plotted markers in three dimensions at the highest system sampling rate (100 Hz) turned out to be suitable for the implementation of analytical and visual kinematics biofeedback. An example of the achievable level of complexity in terms of marker disposition model and graphic representation is reported by describing a demonstration of the real-time representation of human face movements. A clinical application of the method for patient position definition and control at radiotherapy units is presented. PMID- 10367460 TI - Development of a non-invasive treatment system for urinary incontinence using a functional continuous magnetic stimulator (FCMS). AB - A system composed of a functional continuous magnetic stimulator (FCMS) and a saddle-type coil has been developed for non-invasive treatment of urinary incontinence, especially stress incontinence and urge incontinence. The FCMS conditions were as follows: 2 kW maximum electrical power consumption, 800 V maximum capacitor voltage, 720 microseconds pulsewidth (180 microseconds rise time), and 5-30 Hz frequency. A frequency between 5 and 10 Hz is used to treat urge incontinence and a frequency between 25 Hz and 30 Hz is used to treat stress incontinence. The coil (120 mm long, 90 mm wide and 50 mm thick) fits the most suitable region for this treatment, the region from the anus to the perineum. The coil is cooled to maintain a coil temperature between 20 and 25 degrees C so that it can be used efficiently and safely. In experiments with anaesthetised dogs, it was confirmed that the urethral pressure increased when the circumference of the perineum received continuous magnetic stimulation of 720 microseconds pulsewidth (180 microseconds rise time), 10 Hz frequency and about 520 V capacitor voltage. This result suggests that magnetic stimulation can be effective as a urinary incontinence therapy. PMID- 10367462 TI - Neural network technique for detecting emergency states in neurosurgical patients. AB - The problem of reliable detection of life-threatening situations in the neurosurgical patient undergoing treatment in the ICU is still far from reaching a satisfactory solution, although several methods of clinical and instrumental evaluation have recently been developed for the early detection of oncoming signs of danger. Continuous monitoring of intracranial pressure (ICP) provides neurosurgeons with valuable information about the current condition of the patient. However, it is increasingly felt that traditional methods of extracting information from the ICP signal have reached their natural limits, mostly because of difficulties in fitting the appropriate mathematical model to this non-linear and non-stationary process. Successful implementations of artificial neural networks in many medical tasks have encouraged the application of this method of ICP processing. Two problems are considered: the prediction of trends in ICP, and recognition of the configuration of unfavourable symptoms likely to signal danger for the neurosurgical patient. The construction of neural network predictors of ICP trends is based on wavelet pre-processing of the original signal. The approach to the second task involves pre-processing of the ICP with spectral and statistical methods and classification of the extracted features of the current signal on an arbitrarily selected scale of danger. PMID- 10367461 TI - The volume conductor may act as a temporal filter on the ECG and EEG. AB - The influence of the volume conductor on the EEG, MEG, fetal ECG and fetal MCG is studied by means of simulations. The assumption that the Maxwell equations can be used in a quasi-static approximation is reconsidered and the fact that the conductivity of human tissue is frequency dependent is taken into account. It is found that displacement currents have a substantial effect on the fetal ECG and to a lesser degree on the fetal MCG. Moreover, the frequency dependence of the conductivity of the tissues within the head may have a considerable effect on the EEG. PMID- 10367463 TI - Fragmentation of bandpass-filtered QRS-complex of patients prone to malignant arrhythmia. AB - The structure of high-frequency components of electric and magnetic signals from the heart during the depolarisation phase is investigated. After averaging and broadband filtering with a binomial bandpass filter (37 Hz-90 Hz), the fragmentation of the QRS-complex is quantified. The number of extrema M and a new score value S are calculated from the signals of three electrical leads and one magnetic lead of 23 healthy subjects, 23 patients with coronary heart disease (CHD) without reported event of ventricular tachycardia or fibrillation at the time of measurement, and eight patients with CHD who have suffered from malignant tachycardia. For the parameter M, the sensitivity and specificity for healthy subjects against patients with CHD and ventricular tachycardia for the magnetic lead (the best electric lead) are 100% (75%) and 100% (100%). For the magnetic lead (best electric lead) and parameter S, the sensitivity and specificity are 100% (75%) and 95.6% (100%). PMID- 10367464 TI - Pressure measurements during injection of corticosteroids. AB - Corticosteroid injection into the orbit, eyelid and larynx is a common treatment for inflammation and neoplasm. Complications include embolisation into the ocular circulation resulting in permanent loss of vision. The overall aim of the reported research is to develop an injection cannula and monitoring system which can prevent inadvertent embolisation into the ocular circulation during injection of corticosteroids. To that end, a special cannula was designed that allows simultaneous estimation of pressure at the tip of the cannula and flow rate during injection. The cannula was tested with backpressures corresponding to physiological ranges of 0 to 125 mmHg and injection flow rates of 3 to 11 cm3 min 1. The estimated pressure at the tip of the cannula during injection of corticosteroids was compared with direct pressure measurements. The results show that the mean estimated pressure is linearly related to the mean measured pressure with a slope of 0.99 and correlation coefficient of 0.99. Statistical analyses show that with standard error of estimate (SEE) of 2.14 mmHg, the estimated pressure is well within the 95% prediction interval limits of the measured values. The estimation of pressure from the cannula and monitoring system was accurate and warrants further testing in animal models. PMID- 10367465 TI - Whole body urea kinetics. AB - Improved methods are needed for dose quantification in dialysis because the kinetic modelling based on blood side urea concentrations, as currently used, is subject to several theoretical and practical problems. Since the treatment induces a disequilibrium, it is necessary to make assumptions about the distribution and exchange of urea within the body. A new method is presented which avoids these problems. The amount of urea removed is determined from continuous measurements in the spent dialysate. It is shown that the relative dialysis efficiency K/V can be calculated from the mass removal rate in relation to the mass remaining in the body, which is also determined from the measurements. As a by-product the total mass of urea in the body before the treatment is obtained. This can be used together with the blood concentration to calculate the urea distribution volume, which might be used as an additional clinical parameter. The new method was tested in three in vitro treatments of a container with a known amount of urea added to a known volume of dialysate. The calculated dose KT/V and initial urea mass differed from the true values by less than 3%. PMID- 10367466 TI - Multichannel laser-Doppler probe for blood perfusion measurements with depth discrimination. AB - The laser-Doppler method is frequently used in clinical experiments for blood perfusion measurement. A multichannel laser-Doppler probe was constructed, and calibrated and evaluated using a flow model and real examination of healthy subjects. Results obtained on a three-capillary flow model suggest that the probe can be used to obtain information from superficial and deeper layers of the tissue. The influence of the optical arrangement of the laser-Doppler probe on the parameters of post-occlusive reactive hyperaemia was investigated in a group of 15 healthy subjects. A statistically significant increase of biological zero with distance between emitting and detecting fibres was noted. PMID- 10367467 TI - Non-invasive studies of peripheral vascular compliance using a non-occluding photoplethysmographic method. AB - A non-invasive technique is implemented to measure a peripheral vascular compliance index Cindex, using an infrared photoplethysmographic waveform as an indicator of intravascular volume change and a continuous blood pressure monitor to measure the blood pressure during each heart-beat. The non-linear behaviour of Cindex with pressure and the effect of age on Cindex are studied in 62 males (15 73 years). Repeatability tests and the effect of ice-water exposure of a portion of a limb are studied in 10 and 14 subjects, respectively. For each individual, Cindex measurements are taken at discrete values of local mean arterial pressure (Pmean), and a Cindex against Pmean plot is obtained. There is a statistically significant difference (p < 0.05) in Cindex for the lower values of Pmean (60-100 mmHg) between two age groups formed (15-52 and 58-73 years). The cold-pressor test (CPT) shows a 68% median decrease in Cindex, with an inter-quartile range of 60-77%, in a matter of seconds. The results suggest that Cindex may be a useful noninvasive indicator of peripheral vascular compliance in humans. PMID- 10367468 TI - Staffieri tracheo-oesophageal prosthesis for voice rehabilitation after laryngectomy: an evaluation of characteristics. AB - Experimental results on voice prostheses used for the rehabilitation of patients that have lost their vocal function after total laryngectomy are presented. The purpose is to evaluate the difference in aerodynamic behaviour between Staffieri voice prosthesis and other commercial valves (Groningen standard, Groningen low pressure, Panje, Provox). Two different equipments for flow-rate measurement were designed and built to compare the performance of the valves. The valves have been experimentally tested under different conditions of airflow through the valve and tracheal side pressure. The data allow calculation of the airflow resistance, the parameter usually used to compare the performance of valves. The valves have also been experimentally tested under different conditions of fluid flow through the valve and oesophageal side pressure (reverse flow). Comparing the airflow resistance of Staffieri valves of different length L and different angular extension of the razor-thin silt alpha, it has been observed that the parameter alpha has a significant influence on the characteristics, while the effect of the length L is negligible. The airflow resistance of the Provox, Groningen low pressure and Staffieri alpha = 270 degrees valves are comparable; the Panje and Staffieri alpha = 180 degrees have similar behaviour; while the Groningen Standard is comparable to the Staffieri alpha = 90 degrees. Regarding reverse flow, it is pointed out that for most of the valves (Staffieri and commercial valves), at different oesophageal pressures the fluid flow is smaller than the flow that can be tolerated by patients without giving problems. PMID- 10367469 TI - A wideband high common mode rejection ratio amplifier and phase-locked loop demodulator for multifrequency impedance measurement. AB - Design considerations and implementation of a multifrequency measuring channel for application in the field of bio-impedance measurement are discussed in this paper. The input amplifier has a differential configuration which is electrically isolated from the remaining circuits. Transformer coupling provides improved common mode rejection when compared to non-isolated input stages. The frequency characteristic of the section between input and demodulator is flat within +/- 0.1 dB between 4 kHz and 1024 kHz. The synchronous demodulator is based on a wideband switched video amplifier. In contrast to commonly used lock--in techniques, the carrier for demodulation is recovered from the input signal by means of a phase-locked loop. This method ensures zero phase shift with respect to the input signal and improves the accuracy of measurement. The system has been developed primarily for thoracic impedance cardiography (TIC) but has also successfully been applied in the field of total body bio-impedance analysis (BIA). At present an electrical impedance tomograph is under development based on the instrumentation described. Results regarding the measurement range and accuracy are given and some recordings of patient data are shown. PMID- 10367470 TI - Outlining the prostate boundary using the harmonics method. AB - In a computerised ultrasound image guidance for automated prostatectomy system, it is necessary to identify a smooth, continuous contour for the prostate (boundary) from the ultrasound image. The radial bas-relief (RBR) method, which has been reported previously, can extract a skeletonised image from an ultrasound image automatically. After this process the prostate boundary is clearly revealed. However, analysis of the image is far from complete, as there are many spurious branches that create too much ambiguity for the system to define the actual boundary. There are also sections missing from the prostate boundary. Therefore further post-processing is required to describe and define the prostate boundary. In the paper, the harmonics method is used to describe the prostate boundary. The harmonics method uses Fourier information for noise removal and encodes a smooth boundary. The results of using the harmonics method after application of the RBR method on ultrasound images are presented. Factors that affect the performance are also highlighted and discussed. PMID- 10367471 TI - Use of diaphragm transducers in the measurement of pressures on soft materials. AB - Diaphragm pressure transducers are designed to measure pressures in fluids, but have also been applied to measuring pressures on soft materials, such as at the interface between the residual limb of a lower-limb amputee and the supporting surface defined by the prosthetic socket. The reliability and accuracy of Kulite XTM-190 transducer as a pressure monitor on soft materials, such as silicone and Pelite was evaluated in three physical model set-ups. The evaluations included the uniform loading of solid disks of silicone and Pelite, the application of air pressure to the core of a contained thick-walled cylinder made of silicone, and the dynamic indentation of a contained solid silicone cylinder. Sensor measurements in all situations were similar to analytical, iterative or finite element solutions when certain conditions were met. These conditions include: (i) lubricating the interface between the soft material and the supporting structure; (ii) calibrating the transducers under surface and material conditions used during measurements; and (iii) using compatible soft materials (e.g. silicone but not Pelite). PMID- 10367473 TI - Skin substitutes from cultured cells and collagen-GAG polymers. AB - Engineering skin substitutes provides a potential source of advanced therapies for the treatment of acute and chronic wounds. Cultured skin substitutes (CSS) consisting of human keratinocytes and fibroblasts attached to collagen glycosaminoglycan substrates have been designed and tested in preclinical and clinical studies. Cell culture techniques follow general principles of primary culture and cryopreservation in liquid nitrogen for long-term storage. Biopolymer substrates are fabricated from xenogeneic (bovine) collagen and glycosaminoglycan that are lyophilised for storage until use. At maturity in air-exposed culture, CSS develop an epidermal barrier that is not statistically different from native human skin, as measured by surface electrical capacitance. Preclinical studies in athymic mice show rapid healing, expression of cytokines and regulation of pigmentation. Clinical studies in burn patients demonstrate a qualitative outcome with autologous skin that is not different from 1:4 meshed, split-thickness autograft skin, and with a quantitative advantage over autograft skin in the ratio of healed skin to biopsy areas. Chronic wounds resulting from diabetes or venous stasis have been closed successfully with allogeneic CSS prepared from cryopreserved skin cells. These results define the therapeutic benefits of cultured skin substitutes prepared with skin cells from the patient or from cadaver donors. Future directions include genetic modification of transplanted cells to improve wound healing transiently or to deliver gene products systemically. PMID- 10367472 TI - Cultivation of human keratinocyte stem cells: current and future clinical applications. AB - Cultured human keratinocytes have a wide spectrum of clinical applications. Clinical results reported by several investigators are, however, contradictory. In this review, the authors discuss the biological and surgical issues which play a key role in the clinical outcome of cultured epidermal autografts used for the treatment of massive full-thickness burns. The importance of cultivation of epidermal stem cells and of their transplantation onto a wound bed prepared with donor dermis is emphasised. The paper also reviews recent data showing that: (i) cultured epidermal autografts bearing melanocytes can be used for the treatment of stable vitiligo; (ii) keratinocytes isolated from other lining epithelia, such as oral, urethral and corneal epithelia, can be cultivated and grafted onto patients suffering from disabling epithelial defects; (iii) keratinocyte stem cells can be stably transduced with retroviral vectors and are therefore attractive targets for the gene therapy of genodermatoses. PMID- 10367474 TI - Tissue-engineered human skin substitutes developed from collagen-populated hydrated gels: clinical and fundamental applications. AB - The field of tissue engineering has opened several avenues in biomedical sciences, through ongoing progress. Skin substitutes are currently optimised for clinical as well as fundamental applications. The paper reviews the development of collagen-populated hydrated gels for their eventual use as a therapeutic option for the treatment of burn patients or chronic wounds: tools for pharmacological and toxicological studies, and cutaneous models for in vitro studies. These skin substitutes are produced by culturing keratinocytes on a matured dermal equivalent composed of fibroblasts included in a collagen gel. New biotechnological approaches have been developed to prevent contraction (anchoring devices) and promote epithelial cell differentiation. The impact of dermo epidermal interactions on the differentiation and organisation of bio-engineered skin tissues has been demonstrated with human skin cells. Human skin substitutes have been adapted for percutaneous absorption studies and toxicity assessment. The evolution of these human skin substitutes has been monitored in vivo in preclinical studies showing promising results. These substitutes could also serve as in vitro models for better understanding of the immunological response and healing mechanism in human skin. Thus, such human skin substitutes present various advantages and are leading to the development of other bio-engineered tissues, such as blood vessels, ligaments and bronchi. PMID- 10367475 TI - Pigmented human skin equivalent--as a model of the mechanisms of control of cell cell and cell-matrix interactions. AB - The melanin pigment system in human skin is extraordinarly well developed and assures the photoprotection of the skin against harmful solar radiation. Specific cell-cell interactions between one melanocytes and keratinocytes play a fundamental role in the regulation of melanogenesis and melanin pigementation, the two key elements of this system, giving rise to the concept of a structural, functional collaborative 'epidermal melanin unit,' Early experiments strongly suggested that melanocyte growth and differentiation are regulated by paracrine factors from keratinocytes and other skin cells. In addition, co-culture studies with keratinocytes has shown that the extracellular matrix acts as a local environmental signal for dendrite formation and melanogenesis. Attempts to reconstruct pigmented human skin in vitro have made great progress over the last decade. The behavior of cells in these pigmented human skin equivalents closely resembles that in vivo, and the cells can still respond to appropriate extrinsic regulatory stimuli such as ultraviolet radiation. Keratinocytes and fibroblasts have been shown to be active partners in the regulation of melanocyte distribution, viability and other differentiation functions, presumably by direct contact and the effects of various soluble paracrine factors. By reproducing cell cell and cell-matrix interactions, these culture systems provide a promising experimental model for investigating regulation of the skin pigmentary system and the role of photoprotection against harmful solar radiation. PMID- 10367476 TI - Reconstructed human epidermis composed of keratinocytes, melanocytes and Langerhans cells. AB - In addition to their basic biological interest, models of reconstructed epidermis provide useful tools for in vitro assessment of the toxicology and efficacy of new chemicals and drugs. The fact that the majority of these in vitro models are composed only of keratinocytes has excluded their use in the fields of skin pigmentation and immunology. After the successful introduction of functional melanocytes into the epidermal reconstruct, the integration of Langerhans cells remains an important challenge, particularly since after isolation of Langerhans cells from human epidermis, these cells cannot be subcultured and do not integrate into the reconstructing epidermis. The authors show that cord blood derived and CD34+ progenitors isolated from the peripheral blood give rise to residential Langerhans cells when co-seeded with normal human keratinocytes. PMID- 10367478 TI - Is serum transferrin receptor a sensitive marker of iron repletion in patients with iron-deficiency anemia and hemodialysis patients? AB - BACKGROUND: Serum transferrin receptor (sTfR) concentration has been recognized as a reliable laboratory indicator of iron deficiency in recent years. But its response to iron supplementation has not been investigated. METHODS: We evaluated the sTfR concentrations in 15 patients diagnosed with iron-deficiency anemia, in 30 patients receiving maintenance hemodialysis (HD) with iron repletion and in 31 healthy controls. The serial changes of sTfR concentration and their correlation with serum ferritin in patients with iron deficiency under iron repletion were also examined in three patients. RESULTS: In patients with iron-deficiency anemia, the sTfR concentration was 5.6 +/- 2.4 mg/ml, significantly higher than that in the control group (1.8 +/- 0.4 mg/ml) and patients receiving maintenance HD with iron repletion (1.7 +/- 0.5 mg/ml). The three patients with iron deficiency anemia who received eight to 16 weeks of iron supplementation showed steady and significant decreases in sTfR concentration and significant increases in serum ferritin and transferrin saturation. However, the decreases in sTfR concentration did not occur immediately, as did the increases in serum ferritin and transferrin saturation, following iron repletion. There was a four-week delayed response in the decrease of sTfR concentrations as measured against serum ferritin and transferrin saturation. CONCLUSIONS: sTfR concentration may not be as effective as an early index of iron repletion compared with serum ferritin and transferrin saturation. PMID- 10367477 TI - Applications of reconstructed skin models in pharmaco-toxicological trials. AB - The development of new cosmetic formulations requires precise assessment of their safety and efficacy. Today, legislation demands quality control combined with severe safety measures, as well as a limited use of animals for such testing (European Community directive 93/35/EEC). Consequently, safety assessment protocols are oriented towards in vivo tests on human volunteers and in vitro alternative methods to animal use, especially tissue engineered skin substitutes. In this paper, dermal and skin equivalents developed in the laboratory are described. The applications of reconstructed epidermis and skin substitutes for pharmaco-toxicological trials are also discussed. These tissue models have been shown to be very useful tools to assess cutaneous irritation, phototoxicity, photoprotection and to perform efficacy tests of cosmetic molecules and finished products. In conclusion, the authors are confident that these in vitro models can contribute to reduce animal use for routine toxicity testing. PMID- 10367480 TI - Arylamine N-acetyltransferase: a possible promoter in Helicobacter pylori-related gastric carcinogenesis. AB - BACKGROUND: The hypothesis of an association between peptic ulcer and infection by Helicobacter pylori in the gastroduodenal tract was suggested by Marshall and Warren in 1984. H pylori infection of the stomach is the most frequent infection in the world and exhibits an age-dependent increase. However, only a very small percentage of those infected develop gastric carcinoma, suggesting that H pylori acts as a cofactor in the pathogenesis of gastric carcinoma. N-Acetyltransferase (NAT) is expressed in uroepithelial cells and colon cytosol, while cytosolic acetyltransferase plays a critical role in susceptibility to arylamine-induced bladder and colon cancer. The presence of NAT activity in H pylori has yet to be determined. METHODS: NAT activity in H pylori from patients with peptic ulcer was studied using an acetyl coenzyme A (AcCoA) recycling assay and high-pressure liquid chromatography with p-aminobenzoic acid and aminofluorene substrates. RESULTS: The NAT activities from a number of H pylori samples were found to be 0.68 +/- 0.10 nmol/min/10(10) colony-forming units (CFUs) (intact bacteria); and 0.90 +/- 0.22 nmol/min/mg protein (cytosol) for the acetylation of 2 aminofluorene, and 0.63 +/- 0.06 nmol/min/10(10) CFUs (intact bacteria) and 0.72 +/- 0.24 nmol/min/mg protein (cytosol) for the acetylation of p-aminobenzoic acid. CONCLUSIONS: These studies show that H pylori has NAT activity, from which we speculate that the bioactivation of food-borne heterocyclic aromatic amines into genotoxic and carcinogenic products in the stomach is a possible promoter in the pathogenesis of gastric cancer. PMID- 10367479 TI - Plasma concentrations of interferon-alpha in patients with liver cirrhosis: relationship to systemic and portal hemodynamics. AB - BACKGROUND: Nitric oxide (NO) plays an important role in the pathogenesis of the hyperdynamic circulation observed in portal hypertensive states. Interferon (IFN) alpha can stimulate NO formation directly or indirectly via cytokines. However, IFN-alpha concentrations seem to increase or decrease in cirrhotic patients. This study investigated the plasma concentration of IFN-alpha in patients with cirrhosis and its relationship to systemic and portal hemodynamics. METHODS: Thirty-six patients with cirrhosis and 47 healthy controls had blood samples taken for the determination of plasma concentrations of IFN-alpha by enzyme linked immunosorbent assay. Systemic and portal hemodynamics were measured in patients with cirrhosis on the same day of blood sampling using Swan-Ganz catheterization and the thermodilution technique. RESULTS: As compared with healthy subjects, patients with cirrhosis demonstrated a significantly higher IFN alpha detectable rate (> 3 pg/ml, 14.9% vs 36.1%, p < 0.05). In cirrhotic patients, the IFN-alpha detectable rates were similar between those with and without decompensation, a hepatic venous pressure gradient greater than 12 mmHg, or the presence of large esophageal varices (p > 0.05). There was no significant difference in the systemic vascular resistance or hepatic venous pressure gradient between cirrhotic patients with and without a detectable plasma IFN alpha concentration. CONCLUSIONS: Plasma IFN-alpha concentrations tended to increase in patients with cirrhosis. However, IFN-alpha concentrations do not play a role in the hyperdynamic circulation observed in patients with cirrhosis. PMID- 10367481 TI - Body position, membrane diffusing capacity and pulmonary capillary blood volume in chronic bronchitis and pulmonary emphysema. AB - BACKGROUND: The effect of body position on diffusing capacity and its components, membrane diffusing capacity (Dm) and pulmonary capillary blood volume (Vc), in patients with chronic obstructive pulmonary disease (COPD) has remained elusive. This study was designed to evaluate the effect of body position on diffusing capacity for carbon monoxide (DLco), Dm and Vc in male patients with chronic bronchitis and pulmonary emphysema. METHODS: Pulmonary function tests including spirometry and lung volume were assessed in the erect position, and DLco, Dm and Vc were measured in the erect and supine positions in a random order in 17 men with chronic bronchitis and 19 men with pulmonary emphysema. RESULTS: Spirometry results and lung volumes were comparable between both groups of patients; however, significantly lower values of DLco and Kco (DLco corrected by alveolar volume, VA) were observed in the emphysema than in the bronchitis group. In the bronchitis group, Kco and Vc were significantly higher in the supine than in the erect position, but Dm was significantly lower in the supine position. Alternation of body position did not significantly affect DLco and its components in the emphysema group. DLco, Kco and Vc in both the erect and supine positions were significantly higher in the bronchitis than in the emphysema group. Vc-SE (SE, the data in the supine minus those in the erect position) was also significantly higher in the bronchitis group. In the bronchitis group, DLco-SE was significantly correlated with Dm-SE and Vc-SE. However, Kco-SE was highly correlated with Dm-SE. In the emphysema group, DLco-SE and Kco-SE were highly correlated with Vc-SE only. CONCLUSIONS: An increase in Vc in the supine position may account for the postural effect on Kco in bronchitis patients. In patients with pulmonary emphysema, decreased DLco and an absence of postural effect on DLco and its components may be due to a widespread abnormality of the pulmonary capillary bed. These findings may be of value in elucidating the difference in mechanisms of impaired gas exchange between patients with chronic bronchitis and pulmonary emphysema. PMID- 10367482 TI - Influence of Helicobacter pylori on gastric secretion and gastrin release in normal Chinese subjects. AB - BACKGROUND: Controversy exists concerning the influence of Helicobacter pylori on gastric secretion. Hyper-, normo- and hyposecretion of gastric acid in normal subjects with H pylori infection have been reported, although there is no such report for Chinese subjects. The goal of this study was to identify the effect of H pylori on gastric secretion in normal Chinese subjects. METHODS: Twenty normal subjects with a normal upper gastrointestinal tract by endoscopy were recruited. H pylori status was assayed by a rapid urease test. Gastric secretion and gastrin release were also measured. RESULTS: Among the subjects studied, nine were infected with H pylori. All enrolled subjects were males. Age and body weight were similar between both groups. No significant difference was found in basal acid output, maximal acid output, basal pepsin output or maximal pepsin output between the H pylori-positive group (median, 1.1 mmol/hour, 95% confidence interval 0.2-3.6 mmol/hour; 8.0, 3.0-18.3 mmol/hour; 0, -1.3-11.2 mmol/hour; and 4.1, -4.2-59.3 mmol/hour, respectively) and the H pylori-negative group (2.5, 1.3-11.3 mmol/hour; 12.2, 8.7-26.9 mmol/hour; 4.3, 1.8-13.5 mmol/hour; and 14.8, 5.7-73.0 mmol/hour, respectively). Serum basal gastrin and pepsinogen I concentration were 63.5, 50.0-78.6 pg/ml and 75.1, 50.6-89.8 ng/ml in the H pylori-positive group, and 65.9, 50.2-79.8 pg/ml and 79.1, 59.5-120.1 ng/ml in the H pylori-negative group (p > 0.05). CONCLUSIONS: H pylori plays no role in the gastric secretion and gastrin release in normal Chinese subjects. PMID- 10367483 TI - Traumatic diaphragmatic hernia with delayed presentation. AB - BACKGROUND: Traumatic diaphragmatic rupture may occur in patients with thoracoabdominal injuries, and still poses a diagnostic challenge to surgeons. Those patients who survive the events without being diagnosed in the acute phase develop chronic traumatic diaphragmatic hernia. In this study, we reviewed cases of chronic traumatic diaphragmatic hernia managed over the past 35 years. METHODS: We retrospectively evaluated the clinical courses and radiologic images of 24 cases with chronic traumatic diaphragmatic hernia. RESULTS: Motor vehicle accident with blunt abdominal trauma was the most important mode of injury. Herniation was more common to the left plural cavity than to the right. The interval between injury and the onset of symptoms ranged from two weeks to 40 years (average, 7.3 years). Vague chest pain, shortness of breath, and bowel obstruction are the most common presentations. Chest radiographic findings suggested the diagnosis of diaphragmatic hernia in 20 patients. Barium study of the gastrointestinal tract was required to confirm the diagnosis. The most common herniated abdominal viscera were the stomach and colon. All patients received thoracotomy with reduction of hernia organs and closure of the diaphragmatic defect. The hospital course was uneventful with no operative mortality. CONCLUSIONS: Careful interpretation of radiographic images and early surgical intervention are essential in the management of patients with chronic traumatic diaphragmatic hernia. Thoracotomy with reduction of herniated organs can be performed safely with satisfactory results. PMID- 10367484 TI - In vitro activities of macrolides against gram-positive aerobes, Haemophilus influenzae, Moraxella catarrhalis and Bacteroides fragilis in Taiwan. AB - BACKGROUND: New macrolides with improved pharmacokinetic characteristics have been developed and introduced for clinical use in Taiwan. In order to understand the antibacterial activities of these new macrolides, we tested their in vitro activities against common pathogenic bacteria. METHODS: Minimum inhibitory concentrations (MICs) of azithromycin, clarithromycin, dirithromycin, erythromycin and roxithromycin for clinical isolates collected from six teaching hospitals in Taiwan were determined by the agar dilution method. The tested bacteria included methicillin-sensitive and -resistant coagulase-negative staphylococci, methicillin-sensitive and -resistant Staphylococcus aureus, viridans streptococci, Streptococcus pneumoniae, Streptococcus pyogenes, Enterococcus spp, Corynebacterium spp, Haemophilus influenzae, Moraxella catarrhalis and Bacteroides fragilis. RESULTS: High MICs were detected against most of the bacteria tested except for H influenzae and M catarrhalis. The MIC90 for viridans streptococci, S pneumoniae, S pyogenes, Enterococcus spp, S aureus (both methicillin-sensitive and -resistant), coagulase-negative staphylococci (both methicillin-sensitive and -resistant), Coryne-bacterium spp, and B fragilis were all at least 128 micrograms/ml. Wide ranges of MICs were demonstrated. CONCLUSIONS: Most bacteria tested were highly resistant to macrolides. This result is a warning for clinicians that rational use of antibiotics, including macrolides, is mandatory. PMID- 10367485 TI - Iatrogenic abdominal scar endometriosis: a case report. AB - We report the case of a patient with abdominal scar endometriosis following a cesarean section. The rarity of this localization and its appearance on computerized tomography is shown. We emphasize the combination of history and image study of this pathology in the differential diagnosis of other diseases. PMID- 10367486 TI - Primary Salmonella iliopsoas abscess: a case report. AB - Primary iliopsoas abscesses are usually hematogenous or seeded via the lymphatic system from an occult focus. Staphylococcus aureus has been reported to be the predominant pathogen, whereas Salmonella sp has rarely been reported to be a major pathogen. We report the case of a 63-year-old woman who presented with a prolonged fever of two weeks' duration. On admission, physical examination revealed tenderness over the left lower abdomen and hip joint, with her thigh in constant flexion. Computerized tomography of the abdomen revealed an iliac fossa abscess. The drained pus culture yielded Salmonella group B. Percutaneous catheter drainage and appropriate antimicrobial therapy with ciprofloxacin eventually yielded good results. There was no evidence of other underlying diseases predisposing the patient to the formation of iliopsoas abscess. Salmonella infection should be considered in the diagnostic protocols of iliopsoas abscess in Taiwan, where salmonellosis is prevalent. PMID- 10367487 TI - Using three-dimensional-computerized tomography as a diagnostic tool for temporo mandibular joint ankylosis: a case report. AB - Roentgenographic examination has long been a useful diagnostic tool for temporo mandibular joint (TMJ) disease. The methods include TMJ tomography, panoramic radiography and computerized tomography (CT) scan with or without injection of contrast media. Recently, three-dimensional CT (3D-CT), reconstructed from the two-dimensional image of a CT scan to simulate the soft tissue or bony structure of the real target, was proposed. In this report, a case of TMJ ankylosis due to traumatic injury is presented. 3D-CT was employed as one of the presurgical roentgenographic diagnostic tools. The conventional radiographic examination including panoramic radiography and tomography showed lesions in both sides of the mandible. CT scanning further suggested that the right-sided lesion was more severe than that on the left. With 3D-CT image reconstruction the size and extent of the lesions were clearly observable. The decision was made to proceed with an initial surgical approach on the right side. With condylectomy and condylar replacement using an autogenous costochondral graft on the right side, the range of mouth opening improved significantly. In this case report, 3D-CT demonstrates its advantages as a tool for the correct and precise diagnosis of TMJ ankylosis. PMID- 10367488 TI - Spinal cord sarcoidosis presenting as an intramedullary mass: a case report. AB - A case of spinal cord sarcoidosis mimicking an intramedullary tumor is reported because of its rarity and difficulty in diagnosis before surgery. A 66-year-old woman began to suffer from chronic thoracic myelopathy in August, 1996. She had a history of intrathoracic sarcoidosis with left hilar adenopathy in 1991, which disappeared completely after steroid therapy for one month. Magnetic resonance imaging of the T-spine performed on 4 June, 1997, showed normal appearance in T1 weighted and T2-weighted images but abnormal enhancement at the T10-11 level. Open biopsy revealed noncaseating granulomatous inflammation and perivascular lymphocytic infiltration. Following biopsy, methylprednisolone 750 mg was given daily for three days followed by prednisolone 60 mg per day. The patient was discharged on 10 July, 1997, in a stable condition. She died on 22 July, 1997, at a local hospital due to urinary tract infection with sepsis. PMID- 10367489 TI - Postmodernism, the biocultural paradigm, and consciousness. PMID- 10367490 TI - Collaborative physician-patient planning and professional liability: opening the legal door to unconventional medicine. AB - This article explores the possibility of using collaborative planning between physicians and patients to minimize disputes and litigation regarding complementary and alternative medicine. It argues that collaborative planning changes the context for understanding professional liability, moving it from tort concepts that rest upon standards of care to a contract model that rests upon the doctrine of assumption of risk. The article also suggests that the premises of mutual decision making can benefit conventional medical practice in general. PMID- 10367491 TI - Some practical and ethical questions. PMID- 10367492 TI - Opening the legal door to unconventional medicine or opening the legal door to a new era in healthcare? PMID- 10367493 TI - Differences between acute and chronic illness may clarify issues of orthodox versus alternative medicine and tort law versus collaborative planning. PMID- 10367494 TI - A comment on the "collaborative planning" model. PMID- 10367495 TI - The necessary subjectivity of bodymind medicine: Candace Pert's molecules of emotions. AB - This article discusses the relevance of Candace Pert's Molecules of Emotions (1997) to the development of bodymind medicine and argues that Pert's research and conceptual analysis provide the missing link connecting the messages of the mind to physiological effects in the body. The research and analysis also present a fundamental challenge to both the reigning body of medicine and the scientific commitment to so-called objectivity by positing a human organism that can act upon itself with thoughts and feelings--that is, through subjectivity. The contention is twofold: The only adequate medical science is one in which the patient's subjectivity is written into the health and disease equation. But for this to happen the classical/modern concept of science needs to be dramatically reconfigured. PMID- 10367496 TI - On a practitioner's "being" as the true healing agent. AB - The healing power of a doctor comes not just from professional competence but from what the doctor is as a person. PMID- 10367497 TI - Reclaiming the connectedness of medicine. AB - The crisis in medicine is an internal crisis and offers physicians the opportunity to look within for meaning, integrity, and joy in work. PMID- 10367498 TI - Buddhism and medicine: reflections. AB - The author, whose spiritual practice combines Christianity and Zen mediation, explains how meditation and Buddhist perspectives affect her work. PMID- 10367499 TI - Do entheogen-induced mystical experiences boost the immune system? Psychedelics, peak experiences, and wellness. AB - Daily events that boost the immune system (as indicated by levels of salivary immunoglobulin A), some instances of spontaneous remission, and mystical experiences seem to share a similar cluster of thoughts, feelings, moods, perceptions, and behaviors. Entheogens--psychedelic drugs used in a religious context--can also produce mystical experiences (peak experiences, states of unitive consciousness, intense primary religious experiences) with the same cluster of effects. When this happens, is it also possible that such entheogen induced mystical experiences strengthen the immune system? Might spontaneous remissions occur more frequently under such conditions? This article advances the so called "Emxis hypothesis"--that entheogen-induced mystical experiences influence the immune system. PMID- 10367500 TI - Psychosomatic research and the theory of science. AB - In terms of current scientific paradigms, absolute knowledge in the understanding of science is beyond the capacity of the human mind. Today, linear and dualistic science are struggling against emerging new paradigms such as psychosomatic medicine, which threaten the established medical truths of contemporary society. PMID- 10367501 TI - Breast cancer, tamoxifen, and music therapy. PMID- 10367502 TI - The new immunosuppressive era. AB - Immunosuppression, although necessary to enable the graft to escape the consequence of immune surveillance carries some risks for the patients. There is an associated increase in neoplasm, opportunistic infections, and end-organ toxicity. In addition, even with excellent patient compliance, rejection (acute and chronic) remain a major limitation that contributes to the loss or decrease in the function of the allograft. New drugs have been added to the armamentarium of immunosuppressive agents to suppress allograft rejection and to rescue graft from cyclosporine-resistant. Most of the immunosuppressive agents in use today are direct at the T lymphocyte, non-specifically inhibiting T-cell activation and proliferation. The authors described here an update of these new immunosuppressive agents and new strategies used in solid organ transplantation. PMID- 10367503 TI - [Colloid cysts in the 3rd ventricle: a microsurgical series and new perspectives]. AB - Colloid cysts are rare benign CNS lesions (0.5-2% of tumors), mostly located within the third ventricle. Although sometimes asymptomatic they may cause life threatening complications and sudden death and therefore require active treatment in the vast majority of cases. Microsurgical removal warrants excellent radicality at low morbidity/mortality rates but emerging neuroendoscopic techniques have been applied successfully worldwide although longer follow-up of these patients is needed. We present out personal microsurgical series from last decade and discuss the results in the light of current knowledge and preliminary neuroendoscopic experiences. PMID- 10367504 TI - [Substernal goiter: a diagnostic and therapeutic problem. (Report of 39 surgically treated cases)]. AB - Substernal goiter is a rare pathology and its definition varied from author to author. We considered substernal the goiter that has the larger diameter below the thoracic inlet, in accord to definition of Valdoni (1957). The authors examined retrospectively 40 cases of endothoracic goiter, that represent 6.1% of patients operated on thyroid. Surgical access was in 35 cases a cervical collar incision; in 2 patients was necessary to associate a median superior sternotomy and in another patient a lateral thoracotomy. In 3 patients the access was a median sternotomy and in the last patient a posterolaterally thoracotomy. The authors carried out 12 total thyroidectomy (30.7%), 15 subtotal thyroidectomy (38.6%), 10 total lobectomy (25.6%), 2 subtotal lobectomy (5.1%). No mortality was observed. The morbility was represented by 5 monolateral temporary palsy of recurrent laringeal nerve, 6 transitory hypocalcemia, 1 tetanic syndrome, 1 transitory arrhythmy. The follow-up was managed on 35 patients (89.7%). We observed 1 definitive hypoparathyroidism, 1 paralysis of recurrent laringeal nerve, 1 mediastinic relapse. One patient was operated on total thyroidectomy 14 years after the first surgery for a differentiated carcinoma. In conclusion the treatment of substernal goiter is surgical. Debating point is the entity of exeresis. We think that the first choice operation is thyroidectomy, done through a cervical collar incision. Occasionally sternotomy or thoractomy are necessary. PMID- 10367505 TI - [In situ ductal carcinoma of the breast: current status and our experience]. AB - INTRODUCTION: Selecting the appropriate treatment strategy for the individual patient with DCIS represents a major challenge to the surgeon treating breast cancer in the 1990's. CASES AND METHODS: In this study 48 "pure" DCIS patients, treated at Surgical Department of Genoa University, have been selected and divided into Van Nuys prognostic groups. New prognostic classification (Van Nuys) defines three distinct and easily recognizable groups, each of which has a different likelihood of local recurrence if treated with breast conservation. RESULTS: Our results confirm that the risk of local recurrence increases in close relation with prognostic classification groups and suggests that different forms of DCIS may require different treatments. PMID- 10367506 TI - [DRG and gastrointestinal surgery]. AB - The diagnosis-related-groups (DRG) is the cost-based system for hospital reimbursement. However, the proceeds does not coincide with the costs. Aim of the study was to identify the profit, which we could gained with 147, 155, 158, 162, 165, 198 gastrointestinal surgery DRG. 30 consecutive patients, undergone to surgery in Clinica Chirurgica of L'Aquila University, had been studied. We had calculated the daily costs of medical and nursing practice, diagnostic tests, drugs, hospitalization, surgical instruments for every patient's therapy. The DRG proceeds had been correlated with the DRG-costs. The "major gastrointestinal surgery" had not profit (147 DRG: anterior resection of rectum = -354428 Pounds, Miles = -94020 Pounds; 155 DRG: total gastrectomy = -1920641 Pounds). On the contrary, "minimal surgery" had good profits (158 DRG: hemorroidectomy with local anestesia = 1469605 Pounds;162 DRG: sutureless groin hernioplasty = 1561200 Pounds; 198 DRG: videolaparochole-cystectomy: 1208807 Pounds). The study seems to demonstrate the disparity of the reimbursement system related to DRG. However, the surgeons, as managers, must employ warily the resources for producing DRG. PMID- 10367508 TI - [Gastric leiomyoblastoma. (Review of the literature in the light of a case)]. AB - Gastric leiomyoblastoma is a rare benign neoplasm, arising from the smooth muscle cell, that may become malignant. The most important symptoms are epigastric pain, sideropenic anaemia and upper GI bleeding, but frequently the diagnostic iter is difficult and definitive diagnosis is made only with histology after laparotomy. A personal case is reported and Literature data are reviewed; stress being laid on the uncertain biological evolution that influence mostly the choice of therapy. PMID- 10367507 TI - [Restoration of digestive continuity after subtotal gastrectomy: comparison of the methods of Billroth I, Billroth II and roux en Y. Randomized prospective study]. AB - The aim of this study was to evaluate functional results after Billroth I, Billroth II and Roux en Y reconstruction in subtotal gastrectomy. MATERIAL AND METHODS: 45 patients were randomised between 1990 and 1995 and stratified in 3 different groups: 15 BI, 15 BII and 15 Roux. They were investigated by EGDS with multiple biopsies and upper gastro-intestinal scintiscanning, to evaluate gastro esophageal reflux (GER) and dynamics of gastric emptying. Besides they answered a questionnaire: "Gastrointestinal Quality of Life Index" (GIQLI). RESULTS: A reflux esophagitis was found in 5 BI, in 7 BII and in 2 Roux (p < 0.001). No gastric lesions were found in 6 BI, in 5 BII and in 12 Roux, (BI vs. Y, p < 0.05; BII vs. Y, p < 0.001). Chronic superficial gastritis was present in 9 BI, in 4 BII and in 3 Roux (BI vs. Y, p < 0.05). Dynamic scintiscan demonstrated the presence of GER in 5 BI and gastric emptying was fast (37' < T 1/2 < 86'), but incomplete (60' residual activity: 49-62%). GER was evident in 7 BII with slow (28' < T 1/2 < 143') and incomplete (60' residual activity: 48-72%) gastric emptying. GER was detected in 2 Roux and radioactive bolus progression in the Roux limb was fast (24' < T 1/2 < 53') and complete (60 residual activity: 42 52%) (BI vs. Y; BII vs. Y, p < 0,001). There was not statistical significance between GIQLI score in the 3 groups. CONCLUSION: The authors affirm the Roux en Y is the technique of choice in subtotal gastrectomy, if compared with BI and BII. PMID- 10367509 TI - [Portal hypertension and enterostomy: a dangerous combination. Colostomy-induced varices as a rare cause of gastrointestinal hemorrhage. Report of a case and review of the literature]. AB - Variceal bleeding from enterostomy in an unusual complication of portal hypertension. The hemorrhage can be managed in most patients with local measures, but if these manouvres fail, surgical procedures may be required. Treatment options include pressure dressings, stomal revision or disconnection, suture or ligation of stomal varices, bowel resection, and porto-systemic shunting. Recently sclerotherapy and percutaneous transhepatic embolization have been reported. A case of ectopic varices in the site of colostomy complicated by recurrent hemorrhage is described and the therapeutic option utilized is discussed with review of Literature to assess the efficacy of the various treatment options. PMID- 10367510 TI - [Acute pancreatitis: case reports from 1992-1997. Analysis of the costs of DRG, role of ERCP in the reduction of hospitalization time]. AB - In the period 1992-1997, 211 cases of acute pancreatitis were observed: 81 grave cases; 130 slight cases. The ERCP and endoscopic sphincterotomy were performed in 84 cases. From the critical revision 2 groups of patients emerged, homogeneous for etiology and clinical classification, the first submitted to ERCP within 72 hours from the entry in hospital; the other after 72 from the hospitalisation. The "t" Students test for small numbers was significant (p < 001) showing a decrease of hospitalisation in the submitted group to ERCP within 3 days. In 42 cases of acute pancreatitis observed in 1997 a study was made about the hospitalised patients' costs. Two homogeneous groups of acute and slight pancreatitis have been considered. The parameters analysed were about the costs of: a) days hospitalisations; b) pharmaceutical expenses; c) hematological examination; d) surgical operation, laparothomic and laparoscopic surgery; e) instrumental examination. The results of the expense have been compared with the obtained DRG, both for each single case and also for homogeneous group of grave and slight pancreatitis. This comparison showed that the obtained gain through DRG is: a) insufficient to cover the expenses of the grave pancreatitis cases; b) clearly positive in the slight cases. This last result has been very important in the final balance; in fact is only for this positive result that final balance was economically favourable. PMID- 10367511 TI - [The timing in differential diagnosis of cystic tumors of the pancreas: clinical report of three cases of cystadenocarcinoma]. AB - The Authors analyze their own twelve-years experience about the Cystic Tumors of the Pancreas, considering the data existing in the literature. In particular, after a description of anatomo-pathological and clinical characteristics, they set out the problems in the differential diagnosis between the Cystadenocarcinomas and the other benign cystic lesions of the Pancreas. The solution of this problem is often reached only during the operation with multiple biopsies. Moreover they pay attention to the therapeutic choices, determined by the symptomatology of the patients and the localization and the histological kind of the lesions. PMID- 10367512 TI - [Benign non-parasitic cysts of the spleen]. AB - The present retrospective study is related to 7 cases of non-parasitic splenic cysts, 5 post-traumatic and 2 true epidermoid. Symptoms of displacement and pressure on adjacent viscera or physical examination showing an enlarged spleen have caused the beginning of diagnostic investigation in some patients, in others the cyst has been incidentally discovered. The young age and the positive history for prior trauma suggest for pseudocyst but they didn't give us absolute value. We have valued the contribution of the different radiological techniques (scintigraphy, US, CT, selective celiac arteriography, percutaneous biopsy) in the diagnosis of these lesions. The CT has shown to be the gold standard but it wasn't able to distinguish the post-traumatic from true splenic cysts. Such diagnosis is often not sure neither thought the histological study since the epithelial lining typical of the true cysts may have partially or completely destroyed by secondary alterations. However it can be observed also in the pseudocysts by proliferation of epithelial cells included in the traumatic hematoma. Surgery is primarily recommended for the prevention of complications as infection, hemorrhage, rupture in both types of cysts. Partial splenectomy according to the anatomic vascular distribution have permitted in 3/7 cases to resect the cyst preserving the functioning splenic tissue avoiding the long-term adverse effects of splenectomy. PMID- 10367513 TI - [Intestinal anastomosis in urology: comparison of methods]. AB - BACKGROUND: Three different intestinal anastomosis were compared during reconstructive urologic surgery; we evaluated time, cost and incidence of complications. MATERIALS AND METHODS: From November 1993 to February 1997, 45 patients (43 males and 2 females) were carried out ileal resection to fashion 30 ileal neobladder, 8 ileal conduit and 7 augmentation ileocystoplasty. The patients were randomized in 3 groups; in the first the intestinal continuity was performed by B.A.R.; the second one was treated by GLA stapling device; the last underwent a manual suture with interrupted stiches double layer (vicryl). RESULTS: The mean follow-up was 18 months. The mean time of canalization was 6.3 days. The complications were: one intestinal subocclusion (group 1); one abdominal wound infection (group 2); one anastomotic leak and one pulmonary embolism (group 3). The mean time of anastomosis was 18, 12 and 39 minutes respectively. The average total cost was 915,000 lire in the group 1; 1,280,000 lire in the group 2; 632,000 lire in the group 3. CONCLUSIONS: We believe that Biofragmentable. Anastomosis Ring (B.A.R.) is more convenient and advisable in reconstructive urologic surgery, for their quickness, effectiveness and final total cost. PMID- 10367514 TI - [Liver resection with the water dissector; preliminary experience of 8 cases]. AB - New methods for the resection of liver parenchyma was developed in order to simplify liver surgery. The object of our study was to test a method using the water dissector, a rather new equipment. Eight patients, four men end four women, have been operated, from March to September 1997, using the water dissector. Age of the patients ranged from 42 to 83 years (mean age 67 years). Indication for surgery was liver metastasis from colon cancer (6 patients), and gallbladder cancer (2 patients). 1 right lobectomy, 1 left side segmentectomy, 2 V and IV segments resection, 1 unitectomy of the VII segment and 2 III and IV segment resections was performed. In the first four patients we used the Pringle manoeuvre (clamping of the liver stalk), while we did not do it in the last four so we could compare both the operation time and the loss of blood with or without this manoeuvre. One patient died of ARDS in the seventh post-operative day, another patient, who underwent a right lobectomy, developed a biliary fistula which healed in the 10 degrees postoperative day. All the seven surviving patients was in good health, with normal liver ultrasonography when checked on the 31/12/1997. Our results show that the water dissector offers the possibility to isolate vascular stalks very easily, both with a posterior ilar approach and with an intraparenchymal approach, making possible a very accurate haemostasis, in such minimizing blood and biliary losses. This method allows the performing of oncologically correct dissections and in the meanwhile the saving of as much healthy parenchyma as possible. PMID- 10367515 TI - [Primary malignant tumor of the 4th duodenal segment at the angle of Treitz. Report of a case]. AB - A case of 34-years old woman with adenocarcinoma of the IVth duodenal segment extended to the angle of Treitz, treated with duodenojejunal segmentary resection, is described. Clinical features and diagnostic strategies are reported. Personal observation compared with Literature confirms the difficulty of an early diagnosis. The most appropriate surgical techniques for the treatment of these particularly and uncommon neoplasms often discovered in advanced stage are discussed. The better prognosis of these adenocarcinomas compared with those of the proximal duodenum (Ist and IInd segments) can be supported by embryological differences currently to be investigated. PMID- 10367516 TI - [A case of carcinoma arising on the duodenal stump after gastric resection]. AB - The duodenal cancers are very uncommon and the carcinomas arising on a duodenal stump following a B II gastric resection are a real rarity. The Aa. present a case of a patient with a cancer arisen on a duodenal stump 24 years after a gastric resection. The patient was operated with a Whipple D.C.P. PMID- 10367517 TI - Ischemic colitis with chronic stenotic evolution. AB - The authors describes a case of ischemic chronic stenosis of colon after aneurysmectomy. They consider eziopatogenesis, clinical aspects and differential diagnosis with other causes of large bowel stenosis. They illustrate on therapeutic sequence after examination of recent literature. PMID- 10367518 TI - [Primary localization of a hydatid cyst in the major dorsal muscle: report of a case]. AB - A case of primary hydatidosis of major muscle dorsal is reported. Hydatid cyst, though known to occur in many areas of the body, is rare in skeletal muscle. Echinococcosis of muscle is caused by the larval stage of Echinococcus granulosus or, rarely, by the more aggressive Echinococcus multilocularis. Although Echinococcus is the most common cause of liver cyst, hydatidosis of muscle appears to be uncommon, as muscle is involved in only 3% of echinococcal infection. We report a case of a 65 year old woman, of rural origin, with an infestation of the major muscle dorsal. We discuss the serologic and instrumental diagnosis, methods and the hypothesis of a primary muscular localization and surgical and clinical management of these atypical lesion. This authors recommend total pericystectomy Postoperative results were satisfying: no recurrence but one was found at follow up. PMID- 10367520 TI - Perspectives of scientific publications of surgery in Italy. Perspectives on the article of M. Rubino and Al. "Types of surgical articles. Comparative analysis of three Italian and three foreign journals". AB - The globalization of the information and the wide use of the English language in the scientific world impose a not avoidable comparison to the scientific Italian journals, but with value indicators that if accepted without criticism could humble the undeniable internal values. In fact such comparison is based certainly on the quality of the contents, but it has already as background parameter the language of publication if the aim is that of an adequate diffusion. The questions that rise from the analysis of the problem are essentially two: if there is still room nowadays for surgical scientific publications in Italian language and if it is correct to accept in a closed box the indicator of international comparison conditioned by the English language, with the consequence that the definite value judgement is entrusted to external opinions. Certainly the Italian journals of surgery must aim both to quality and to their diffusion for reaffirm and validate their cultural rights. The quality depends on the authors and on the surgical environment in which they operate, beyond the engagement of the scientific direction of the journal; the diffusion on the correct employment of the English language, at last of the abstracts, and on the general editorial strategy pointing to encourage and promote the widest coverage of critique. It remains but to bravely loose the knot represented from the proposal of a uncritical acceptance of the indicator of the IMPACT FACTOR with the consequent absolute judge of quality, that fatally depends from an egemonic Anglo-Saxon filter, not exempt from an unavoidable lobbystic imprint. PMID- 10367519 TI - [Type of surgical articles. Comparative study of 3 Italian and 3 foreign periodicals]. AB - BACKGROUND: Medical progress in the world is divulged to peers and colleagues through books and scientific journals. It is thus linked not only to the quality of articles, but also to their type. MATERIAL AND METHODS: The type of articles published in three Italian periodicals (Annali Italiani di Chirurgia; Chirurgia; Minerva chirurgica) and three foreign periodicals published in English (Annals of Surgery; British Journal of Surgery; Surgery, Gynecology & Obstetrics) was compared. The articles were classified according to their potential contribution to medical progress, being divided into two main groups: research papers and simple case series or case reports. Other types of articles (reviews, description of surgical techniques, editorials etc.) were excluded from the study. RESULTS: Both the total number and the percentage of research papers were markedly superior in the three foreign journals with respect to the three Italian periodicals. All the differences were highly significant. CONCLUSIONS: The number of research papers published in Italian periodicals should be increased. Greater attention should be paid to quality, at the expense of quantity, since serious research inevitably takes longer to perform. PMID- 10367521 TI - [The influence of the traction on the wrist in the course of healing of the radius distal part comminuted fractures]. AB - Traction and external fixation are the methods of choice by treating the comminuted fractures of the distal part of the radius. Bringing in two screws into the second metacarpal bone and the radius proximity from the fracture makes possible the traction and fixate the fractured bone splinters. The reposition of the comminuted fractures needs a very strong traction and provides to expand the ligaments and the wrist articular capsula. The aim of our study is to explain the mechanical influence of the external stabilization on the wrist and the wrist function after the immobilisation with the distraction. In 15 patients the examination was performed. The wrist was estimated on the base of clinical examination and rentgenography. Those examinations were performed within the moment of reduction of the dislocation, than 6 weeks and after next 6 weeks. All results are illustrated on graphics and x-ray pictures. The results of our study prove the profitable influence of the traction on the healing of the damaged articular surface in good clinical and functional position. Even a strong traction has in comparison to the control group (healing conservatively or by using various methods of the internal fixation) no influence on the wrist function. PMID- 10367522 TI - [Elbow contracture: causes, management]. AB - Early results after conservative or operative treatment of 15 patients (6 females, 9 males) for contracture of the elbow are presented. The correction achieved has been evaluated by Mayo Elbow Performance Index and by range of flexion and extension movement. Immobilization of the injured elbow should be as short as possible, fixed deformity is an indication for surgery. Resulting contracture of the elbow depends not only on injury severity but on the type of treatment also. One case has been discussed. PMID- 10367523 TI - [The sensitivity and specificity of sonographic examination in detection of rotator cuff tear]. AB - Sonographic examination was performed in 72 patients (74 shoulders) suffering from different shoulder diseases. The results were correlated with findings obtained during subsequent surgery in 57 patients (58 shoulders) and arthrography in 15 cases (16 shoulders). Rotator cuff tear was proved during surgical procedure in 37 patients (38 shoulders). The sensitivity and specificity of ultrasonography in detection of cuff tear was 98.2% and 90% respectively. In case of partial cuff tear ultrasonography had lower sensitivity--50% while specificity was 96.3%. In detection of supraspinatus tendon tear the ultrasound sensitivity was 100% and specificity 95.6%, in case of infraspinatus tear sensitivity was 66.7% and specificity 94% while for subscapularis tear these values were equal 75% and 98.1%. The size of sonographically estimated cuff defect correlated properly with intrasurgical measurement in 76.3% of all cases. PMID- 10367524 TI - [Total knee replacement with modular Kinemax endoprosthesis in osteoarthritis and rheumatoid arthritis]. AB - One to 7 years results (mean 3 years) of modular kinematic knee arthroplasty were compared in rheumatoid (RA, n = 29) and idiopathic (OA, n = 28) arthritis female patients. Comparisons of means showed lower age at surgery (59 yr. v. 66 yr., p = 0.002), lower body mass (66 kg v. 75 kg, p = 0.001) and lower Hospital for Special Surgery scoring (71.7 v. 80, p = 0.05) in RA patients. Two RA knees with prosthesis showed after 2-3 years deterioration of the previously good result. Late infections were indication for removal of the prosthesis in two patients with RA. Two patients of this group died due to complications of the disease. Authors point that knee arthroplasty carries higher risk in rheumatoid than in idiopathic osteoarthritis. PMID- 10367525 TI - [Functional assessment after operative treatment of femoral shaft fracture]. AB - The paper presents results of treatment in 135 patients (aged 16-77 years) with fracture of the femoral shaft treated with one of the four techniques: open intramedullary Kuntcher nailing, Zespol fixation, AO plating and closed locked and unlocked intramedullary nailing. The assessment has been based on authors' own 4-grade, 35-points scale involving 7 parameters. Closed intramedullary nailing proved to the most efficient. No reoperation in this group occurred. The percentage of excellent and good results was the highest--96.4% and such was the score--32.6 points. Open methods (Kuntcher nailing, Zespol fixation, AO plating) rendered 88.2%, 31.7 points and 59% excellent and good results, 26.2 points in group without complication and reoperated one respectively. PMID- 10367526 TI - [Hip Zespol fixator in osteosynthesis for trochanteric fracture]. AB - Over 250 proximal femur fractures (ones) have been treated in Department with hip Zespol fixator since 1987. There were 159 trochanteric (44 stable and 114 unstable) fractures in this group. Mean patient age at operation was 71 years. Thirty-four patients died before bony union had been achieved. One hundred thirteen cases have been reviewed. In 101 patients the fracture healed without complications, in 12 cases reoperation was required due to fixator failure. The study confirms the need for anatomic reduction, proper surgical technique and most of all weight bearing restriction till radiographically confirmed union. PMID- 10367527 TI - [Intraoperative sectional anatomy of congenital absence of tibia type I A]. AB - Intraoperative, angiographic and preparation findings within lower leg and foot of a child with tibial hemimelia type IA according to Jones are presented. Disarticulation at the knee has been performed bilaterally. Dysplasia of the distal femur, absence of the patellae, extensor apparatuses and cruciates were found. Amputated preparation revealed fibrotic band attached via interosseus membrane to the fibula, presence of unidentified muscle attached to the tip of the lateral malleolus, doubled tibialis anterior muscle on one side and accessory extensor digiti II. Talocalcaneal synostosis was bilateral, abnormal cuboids and accessory bones were found. Popliteal artery division was found at the mid-length of the fibula. Unidentified thick nerve passed the lateral ankle and divided at the sole of the foot. PMID- 10367528 TI - [Musculo-skeletal mechanisms of low back pain]. AB - Authors presented in comprehensive form updated knowledge concerning reasons and mechanisms of musculoskeletal back pain nonspecific symptom which in majority of cases, provides only little inside into the nature of the underlying problem. The physician must however find sufficient time to educate the patient as to the nature of this disorder and likely course during treatment. Understanding the source and mechanisms of the patients pain allows the physician and patient to approach the problem rationally with mutual confidence. The basic aim of the physician is exact identification of the anatomic source of pain and exclude the possibility of underlying systemic or malignant process that might threaten the patient's life. The musculoskeletal system consists of the bones and articulations, ligaments, muscles and tendons, that connect and manipulate them within spinal column. All of these tissues are richly innervated and specific biomechanical conditions cause, that lumbar part of the spine and lumbosacral junction are extremely susceptible to direct and indirect affected by a spectrum of reasons causing pain. The pathomechanism of the pain originating within the facet joints, bone and periosteum, paraspinous muscles and tendons, intervertebral disks, posterior longitudinal ligament and dorsal root ganglion was discussed. Although in majority of causes it possible to control localized pain by means of conservative treatment, however in some patients (0.5%-1%) surgery is needed to decompress neurologic structures, restore stability, and eliminate pain. PMID- 10367530 TI - [Hardness of bone tissue in growing rats treated by fluoride of various concentration]. AB - The aim of the study was to investigate the effect of fluoride exposure on both bone fluorine content and bone hardness in rats. Twenty eight 6-weeks-old female rats were divided in four groups. One group served as a control group and received distilled water to drink, and the other three groups received fluoridated water at different doses (8, 30 and 60 mg F-/l). The rats were followed for 6 weeks. Both femoral bones were obtained from each rat, and changes in cortical bone fluoride content and Vickers microhardness in distal femoral diaphysis were assessed. Fluoride concentration in bone increased significantly with the fluoride intake through the water supply. Microhardness values were significantly greater in groups receiving fluoridated water, with maximum values in the 30 mg F-/l treated group. The results of this study show the usefulness of Vickers microhardness tests in examining mechanical changes in cortical bone after experimental exposure to fluoride. PMID- 10367529 TI - [Stress fracture of free vascularized bone grafts]. AB - Three cases of stress fracture within free vascularized bone graft were found among 35 patients with completed treatment. In 32 years old male stress fracture of free vascularized fibular graft used for tibial reconstruction occurred after a dozen of one (operated) leg hops. In 29 years old female operated due to tibial pseudoarthrosis with free vascularized iliac wing graft the fracture appeared after couple of days of intense walking. In both cases fracture occurred after incorporation of the graft, several months after removal of the external fixator. Pain and swelling of the limb increased over couple of days. In the last case of 25 years old male vascularized fibula transferred to the femur fractured in the presence of external fixator one year after surgery in the course of remodeling of the graft. In all cases X-ray revealed transverse fracture line in the middle of the graft. Plaster cast immobilization has been used in one case only. Graft remodeling with marked hypertrophy was found 4 month later. PMID- 10367531 TI - [Diagnostic difficulties in the diagnosis of bipartite scaphoid: 2 case reports]. AB - The case of 2 males (aged 25 and 48) was reported. The bipartite scaphoid was suspected in both. The trial proved one was congenital and the other posttraumatic. The authors applied the criteria suggested by Cotta. The wrist X ray from childhood and MR may be useful in diagnose. PMID- 10367532 TI - [Diagnostic difficulties in primary hyperparathyroidism: a case report]. AB - The authors describe the case of the 56-year old woman suffering from the primary hyperparathyroidismus caused by the adenoma. After the osteosynthesis the pathological fracture of the femur by angle plate, the patient underwent diagnostic, setting the right diagnosis. The exision of the parathyroidal adenoma led to the disappearance of the symptoms of the disease. PMID- 10367533 TI - [A case of thrombosed popliteal artery by method of endovascular pulse spray thrombolysis in the process of multiple fractures of tibia]. AB - A case of team-treated thrombosed popliteal artery caused by blunt limb trauma with open tibia fracture is presented. Thrombosed artery was treated by endovascular pulse-spray method using Angiomed catheter. Implementation of this method allowed us to dissolve thrombus and to retrieve arterial circulation in the limb. In our opinion this procedure was an alternative for traditional surgery. After revascularisation of the limb orthopedic operation on fractured bones was performed using Zespol technique. PMID- 10367534 TI - [The treatment of multi-fragmental fractures of distal radius by external fixators by own construction]. AB - A retrospective analysis of 20 multifragmental fractures of distal radius treated in Clinic of General and Hand Surgery in Szczecin from March 1996 until November 1997 with own constructed fixateur externe showed in 80% very good and good functional results according to the criteria of Gartland and Werley, Castaing. Mean follow-up was 10 months after operation. The 20 patients were on average 56.5 years old. The fixator was removed after five to at least six weeks. An additional osteosynthesis with Kirschner wires was performed in four cases. PMID- 10367536 TI - Gene therapy: where are we going? PMID- 10367535 TI - [The experience in employing reciprocal gait orthoses]. AB - The paper presents the experience of the authors in employing reciprocal gait orthoses in a group of 23 patients age 3-25 years (mean age 7.8 years). The orthoses were indicated in patients with flaccid paresis (17 children with myelodysplasia and 3 patients with traumatic paraplegia) and with arthrogryposis (3 patients). The follow-up period was 6 months to 5 years (mean 2.4 years). The authors discuss the principles of construction and operation of reciprocal gait orthoses and types of patients in whom they are recommended. The principles of learning walking and using the orthosis are also presented. PMID- 10367537 TI - [The role of gemcitabine in the treatment of bladder tumors]. PMID- 10367538 TI - [21st annual San Antonio Breast Cancer Symposium. 12-15 December 1998, San Antonio, Texas]. PMID- 10367539 TI - Single agent 2',2'-difluorodeoxycytidine in the treatment of metastatic urothelial carcinoma: a phase II study. AB - PURPOSE: To evaluate the therapeutic activity and toxicity of gemcitabine in the treatment of metastatic urothelial carcinoma. PATIENTS AND METHODS: Twenty-four consecutive patients with recurrent- and/or metastatic urothelial carcinoma pretreated with first line cisplatin-based chemotherapy were treated with gemcitabine 1000 mg/m2/week intravenously diluted in 250 cc of normal saline as 20 minutes infusion for 3 consecutive weeks followed by a 1 week rest period. Chemotherapy was repeated every 28 days. RESULTS: All enrolled patients were evaluable for objective response accordingly to an intent-to-treat analysis. A complete response was achieved in 1 patient (4%) and a partial response in 6 cases (25%) for an overall response rate of 29% (confidence limits 18%-39%). The median duration of objective responses was 7.4+ months (range 3.0+/12.8). Six patients showed no changes (25%) with a median duration of 4.0 months. A subjective improvement in tumor-related symptoms was reported by all responding patients, and in 3 patients with no change. Six out of 9 patients with symptomatic bone lesions had a subjective improvement with reduction in analgesic drugs consumption. Objective responses were observed at all sites of disease. The median overall survival was 13.0+ months (range 4.0/16.2+). Over a total of 76 cycles (a mean of 3.1 cycles/patient), grade 1-2 leukopenia was seen in 9 patients (37%), grade 1-2 thrombocytopenia in 4 patients (17%), and grade 1 anemia in only 2 cases (8%). Grade 3 leukopenia was seen in 3 cases (12.5%). Grade 4 leukopenia or grade 3-4 thrombocytopenia were not seen. Gastrointestinal toxicity was very mild. CONCLUSIONS: Single agent gemcitabine at the dose of 1000 mg/m2 on a weekly schedule is active, at least in terms of objective response rate and tumor-related symptoms palliation, against pretreated urothelial carcinoma with good tolerability. These results compare favorably with those achieved with the most active drugs such as cisplatin and methotrexate. PMID- 10367541 TI - Age-related changes in blood pressure twenty-four-hour pattern in normotensive subjects of two populations. AB - OBJECTIVES: This study investigates the systolic (S) and diastolic (D) blood pressure (BP) 24-h pattern in normotensive healthy subjects belonging to two populations characterized respectively by a "non-salt culture" (Italian subjects) and a "salt culture" (Japanese subjects) in their dietary salt intake (4-6 g/day in Italians vs 10-12 g/day in Japanese). The comparison was performed by taking into consideration the within-day variability (WDV) and circadian rhythmicity (CR) of BP with respect to age. MATERIALS AND METHODS: Subjects investigated were 862 normotensive healthy subjects (308 Italians and 554 Japanese), stratified by age from 16 to 75 years, who volunteered for a noninvasive BP monitoring in an ordinary day of their life. The SBP and DBP time series were analyzed via conventional parametric statistics as well as chronobiological procedures. RESULTS: The biometric estimates demonstrate that BP changes in its WDV and CR as a function of age in both populations. Despite the difference in their habitual salt intake, the age-related changes in BP WDV and CR result to be almost comparable at the cross-sectional contrasts, giving origin to age-related trends for SBP and DBP which are significantly parallel. CONCLUSIONS: The comparability of BP WDV and CR in the two populations with a substantial difference in salt intake suggests that the normotensive status in human races is realized despite the difference in their habitual salt intake. This implies the ancestral development of mechanism(s) of adaptation to the possible "sodium luxus consumption". Although the adaptive mechanisms which provide a normotensive regimen under different conditions of sodium intake are almost unexplored, the racial adaptation to dietary salt constitutes, however, the initial condition for the cause-effect nexus between dietary salt intake and hypertension in human populations. PMID- 10367540 TI - [Gemcitabine in advanced stage soft tissue sarcoma: a phase II study]. AB - PURPOSE: To assess the activity and toxicity of gemcitabine in locally advanced or metastatic soft tissue sarcoma patients (pts). PATIENTS AND METHODS: Gemcitabine was administered on days 1, 8, 15 every 4 weeks at a dose of 1.000/1.250 mg/m2, respectively, in pretreated or not pretreated pts. RESULTS: Eighteen pts entered this phase II trial; sixteen had been previously treated with anthracyclines and ifosfamide. A partial response was observed in a woman with fibrous malignant istocytoma, whereas in 7 pts the disease remained stable. Median time to progression was 4 months. The treatment was well tolerated. Grade 4 toxicity was not observed. CONCLUSIONS: These results do not suggest that gemcitabine, in the dose and schedule used in this trial, may be of value in the treatment of soft tissue sarcomas. PMID- 10367542 TI - Bowel function in runners after ingestion of sweeteners. AB - OBJECTIVES: This study investigates the bowel function in outdoor runners drinking sweetened solutions both by determination of H2 in exhaled air and asking about intestinal discomfort. MATERIALS AND METHODS: Twenty-two athletes with a mean age of 22 +/- 3 yrs, received 100 ml of a 10% water solution of saccharose, glucose, mannitol, and isomalt respectively, before a 60 min outdoor run training. The trial was repeated on a rest day, one week later. Samples of the exhaled air were obtained at time zero (basal) and then every 15 min for a period of 3 hrs after administration of the solutions. The H2 was detected by breath test. RESULTS: H2 excretion was lower during exercise. Suprabasal increments of breath hydrogen after saccharose load were significantly higher than after ingestion of mannitol (p < 0.05 between 1 and 3 hrs) and isomalt intake (p < 0.05 over 30 min). After glucose load, suprabasal increments of breath hydrogen were higher than after mannitol (p < 0.05 between 1 and 3 hrs) and isomalt (p < 0.001 between 1 and 3 hrs) ingestion. CONCLUSIONS: The intestinal discomfort in runners drinking sweetened solutions seems to be related to different bowel transit time. PMID- 10367543 TI - [Bioethical aspects of Viagra]. AB - Viagra (sidenafil) has specific activity, and well demonstrated efficacy and tolerability, when used in patients with organic erectile dysfunction. Its mechanism of action besides in restoring a compromised function of erection by improving the vasodilation nitrogen oxide-mediated, through the inhibition of cGMP, in the corpus cavernosum. The drug should be used after giving a correct and complete information to physicians by several experts in the field, including the bioethicist, considering the complexity of the problem of sexuality. The bioethical perspectives of the drug should be considered in relation to the principles of totality (therapeutic principle), autonomy, freedom, responsibility, and integrity of medical profession. Viagra is of value both at the clinical level and moral level, provided the therapeutic principle and the principles of responsibility and freedom are respected in an ethic anthropological dimension that accepts the personalistic principle of corporeity. PMID- 10367544 TI - [Dexrazoxane. Current status and prospectives of cardiotoxicity of chemotherapy]. AB - PURPOSE: To evaluate efficacy of dexrazoxane (Cardioxane) in amelioration of chemotherapy induced cardiotoxicity. DESIGN: The most important experimental and clinical studies have been reviewed. RESULTS: Dexrazoxane has been shown to reduce anthracycline-induced cardiotoxicity. The drug is thought to reduce cardiac toxicity by binding to free and bound iron, thus reducing the formation of anthracycline-iron complexes and the generation of free radicals which are toxic to cardiac tissue. The majority of clinical studies has been performed in patients with metastatic breast cancer. It has been shown that dexrazoxane: 1) significantly reduces the overall incidence of cardiac events, irrespective of pre-existing cardiac risk factors; 2) does not seem to affect anthracycline activity; 3) does not increase the incidence of chemotherapy induced non-cardiac toxic effects; 4) reduces cardiac toxicity in pediatric patients given anthracyclines; 5) is cost-effective. CONCLUSIONS: Dexrazoxane is a valuable drug in amelioration of chemotherapy induced cardiotoxicity, both in adult and pediatric patients. Wether it offers protection against late-onset cardiac toxicity in this latter group of patients, remains to be demonstrated. Dexrazoxane should be always used, preferably from the onset of chemotherapy, when it is believed that cumulative doses of doxorubicin > or = 300 mg/m2 or of epirubicin > or = 480 mg/m2 will be reached. PMID- 10367545 TI - [The role of TC and MRI in the identification, characterization and staging of tumors of the spinal vertebrae]. AB - The early detection and characterization of primary and metastatic spinal bone tumors permits early and appropriate surgical or nonsurgical intervention directed toward preserving life and function. The sensitivity and multiplanar capabilities inherent in Magnetic Resonance imaging make it the imaging procedure of choice in detecting and characterizing a spinal bone lesion. Spiral Computed Tomography with multiplanar reconstruction may be a useful supplementary procedure, especially when detailed bony anatomy, particular of the posterior elements, is required for surgical intervention. Plain films play little if any role in modern imaging. The may be used as screening procedures in situations in which MR and CT are not available. PMID- 10367546 TI - [Dyspepsia and Helicobacter pylori]. AB - Since Helicobacter pylori (Hp) was first isolated in 1983, much work has been carried out on the pathogenic effects of this organism. Hp infection is common in humans and currently is the most important etiologic agent in the development of chronic active gastritis, gastric and duodenal ulcers, carcinoma and Malt lymphoma of the stomach. Moreover Hp infection has also been associated with various extradigestive diseases. At present, a role of Hp infection in dyspepsia is discussed. Dyspepsia is defined by persistence of pain, burning or discomfort localised to the upper abdomen; some authors include in dyspepsia symptoms such as belching, bloating, alitosis, nausea, postprandial repletion, vomiting and regurgitation. In absence of any underlying pathologies, such as peptic ulcer, gastroesophageal reflux, pancreatitis, biliary tract disease or others, dyspepsia is defined as functional or idiopathic dyspepsia. Functional dyspepsia may be distinct in ulcer, reflux or dysmotility-like dyspepsia and unspecified dyspepsia. Hp infection is common in dyspeptic patients and a role of this bacterium has been postulated mostly in ulcer-like dyspepsia. Mechanisms by when Hp induces dyspeptic symptoms are uncertain; bacterial cytotoxins, phlogosis mediators, activity of chronic gastritis Helicobacter-related and host immune response probably play an important role in pathogenesis of functional dyspepsia. However, dyspepsia is not present only in infected patients; therefore other pathogenic factors may be implicated in expression of dyspeptic symptoms in uninfected subjects, such as gastric dysmotility, modifications of gastric output or altered visceral sensibility, psychological factors, gastroesophageal reflux and irritable bowel. PMID- 10367547 TI - [Docetaxel in the treatment of metastatic carcinoma of the salivary glands: report of a case]. AB - We report a case history of a patient treated with single agent docetaxel (100 mg/mq every three weeks) for a metastatic salivary gland carcinoma. After surgical treatment of primary tumor, this patient underwent two subsequent resections for local relapses. However, lung and liver metastases developed which were unsuccessfully treated with carboplatin + doxorubicin, and bleomycin. Salvage therapy with docetaxel induced a significant regression (> 50%) of both liver and lung metastases. Docetaxel seems to be active in metastatic salivary gland carcinoma, and a multicentric phase II trial is now ongoing to better evaluate its activity in this disease. PMID- 10367548 TI - [Treatment of malaria in the 19th and 20th centuries]. PMID- 10367549 TI - Comparison of intravenous and intranasal heroin self-administration by morphine maintained humans. AB - Eight heroin-dependent individuals, maintained on divided daily doses of oral morphine, participated in a 2.5-week inpatient study comparing the effects of intranasal (IN) (placebo, 12.5, 25, 50, 100 mg) and intravenous (IV) (placebo, 6.25, 12.5, 25, 50 mg) heroin. Each morning, participants received $20 and a sample dose of heroin, and each afternoon they had the opportunity to self administer all or part of the morning heroin dose or money amount. Participants responded under a modified progressive-ratio schedule (PR 50, 100, 200, 400, 800, 1200, 1600, 2000, 2400, 2800) during a ten-trial self-administration task. During each trial, participants could respond for 1/10th of the heroin dose or 1/10th of the money amount. The total amount of heroin and/or money chosen during the self administration task was given at the end of the task. Thus, participants received drug and/or money twice each day: once during the morning sample session and once during the afternoon self-administration session. Participants received IV solution and IN powder simultaneously during each dosing; only one route contained active drug. Heroin produced dose-related increases in break point values by both routes of administration. Although IV heroin was approximately four-fold more potent than IN heroin, the maximal break point values for both routes were not significantly different. A similar difference in potency between the IV and IN routes was found for several ratings of subjective effects (e.g., "I feel a good drug effect," "I feel high"), but maximal subjective ratings were lower for IN compared to IV heroin. These results suggest that the reinforcing efficacy of heroin is similar by the two routes of administration, but that IN heroin is less potent than IV heroin. The results also underscore the importance of evaluating drug self-administration in the evaluation of the abuse liability of drugs. PMID- 10367550 TI - Naltrexone blockade of nicotine effects in cigarette smokers. AB - RATIONALE: The role of endogenous opiate systems in cigarette smoking remains unclear. In laboratory animals, opiate antagonists block many of the effects of nicotine, but in humans they do not consistently alter smoking behavior. OBJECTIVE: This study explored the effects of naltrexone, alone and in combination with nicotine, on smoking behavior. METHODS: In a double-blind, double-dummy, within-subjects design, 19 regular smokers received four treatments of 1 week duration: naltrexone tablet (50 mg) plus placebo skin patch, placebo tablet plus nicotine skin patch (21 mg/24 h), naltrexone tablet plus nicotine skin patch, and placebo tablet plus placebo skin patch. During each treatment, subjects rated their responses to nicotine-containing and denicotinized cigarettes in the laboratory, and to their own brand of cigarette smoked ad libitum outside the laboratory. RESULTS: Pretreatment with the nicotine patch attenuated smoking-induced decreases in craving, negative affect, and rates of ad lib smoking, and potentiated the aversiveness of a cigarette. Naltrexone reversed these effects of the nicotine patch, and produced negative effects on mood. CONCLUSIONS: The blockade of nicotine's effects by naltrexone supports a role for opioid mechanisms in cigarette smoking. PMID- 10367551 TI - Phenylalanine kinetics are associated with tardive dyskinesia in men but not in women. AB - RATIONALE: An association between tardive dyskinesia (TD) and severely impaired metabolism of the large neutral amino acid (LNAA), phenylalanine (Phe) was defined in a group of mentally retarded patients. Subsequently, an altered kinetics of Phe was associated with TD in men with schizophrenia based on plasma analyses subsequent to the ingestion of a protein meal. METHODS: In the present study, a standardized oral challenge of pure Phe (100 mg/kg in 170 ml orange juice) was administered to psychiatric patients of both sexes (n = 312), with and without TD after an overnight fast. Plasma LNAA levels were assayed both fasting and 2 h subsequent to the ingestion of the challenge. The extent of the increase in plasma Phe levels 2 h following a standardized challenge is determined by the sum of the kinetic processes of plasma absorption, tissue distribution, metabolism and elimination. RESULTS: The study hypothesis, that TD would be associated with significantly higher post-challenge plasma Phe indices of an absolute plasma Phe level and plasma Phe/LNAA ratio (a brain availability measure), was verified for the study men (n = 209), but not for the study women (n = 103). CONCLUSIONS: The demonstrated altered kinetics of Phe in men with TD indicates a greater availability of Phe to the brain in these men. We suggest that the disorder may be related to the effects of this greater availability. Such effects could be the direct neurotoxic effects of Phe and its metabolites and/or the modulating effects of these compounds on the synthesis of the monoamine neurotransmitters. The fact that TD (Yes/No) group differences in post challenge plasma Phe indices were not seen for the study women suggests the possibility of a sex difference in the biology of TD that we propose may be reflective of the young age of the study sample. PMID- 10367552 TI - Branched chain amino acids decrease tardive dyskinesia symptoms. AB - RATIONALE: Prior studies had suggested (a) that a lessened ability to clear ingested forms of the large neutral amino acid (LNAA), phenylalanine (Phe), was associated with having tardive dyskinesia (TD), and (b) that greater availability of a group of LNAA, the branched chain amino acids (BCAA), concomitant with the lower availability of Phe to the brain are associated with a decrease in TD symptoms. The present study was then conducted to test whether increasing the daily intake of the BCAA would decrease the symptoms of TD. METHODS: A 2-week trial of a BCAA medical food administered three times a day was conducted in nine men with long neuroleptic treatment histories. Frequency counts of TD movements were collected by videotape throughout the trial and these tapes were analyzed in blind random sequence for both patient and time for TD symptom level changes subsequent to completion of the trial. Plasma levels of the LNAA were also collected throughout the trial. RESULTS: A statistically significant decrease in the level of TD symptoms was observed for the sample. The symptom changes were also clinically significant in that six of the nine subjects had symptom decreases of at least 58%, with all subjects having a decrease of at least 38%. BCAA administration increased plasma BCAA concentrations and BCAA/LNAA ratios and decreased plasma Phe concentrations and the Phe/LNAA ratio. Analyses indicated a strong significant correlation between the percent increase in the plasma BCAA values at the first administration and the percent improvement in TD over the trial in eight of the nine subjects. CONCLUSIONS: The BCAA show promise as a treatment for TD. The decrease in TD symptoms seen in the trial may have been modulated by the BCAA treatment-induced increased availability of the BCAA and decreased availability of Phe to the brain. PMID- 10367553 TI - Opposite effects of 3,4-methylenedioxymethamphetamine (MDMA) on sensorimotor gating in rats versus healthy humans. AB - RATIONALE: Prepulse inhibition of acoustic startle refers to the reduction in the startle response when the startling stimulus is preceded by a weak prepulse stimulus. This phenomenon provides an operational measure of sensorimotor gating that has been found to be reduced in patients with schizophrenia and rats treated with serotonin agonists or serotonin releasers. OBJECTIVE: In this study, we compared the effects of a serotonin releaser, MDMA, on prepulse inhibition in laboratory rats and healthy human volunteers. In particular, we investigated whether MDMA disrupts PPI in humans as observed in animal studies. METHODS: Rats were tested after placebo and MDMA in a counterbalanced order at an interval of 1 week, with separate groups of rats being used for each dose of MDMA (1.7, 5.4 and 17.0 mg/kg). On each test day, rats were first tested after no injections and retested 2 h later, 10 min after a subcutaneous injection of placebo or MDMA. For the human study, a placebo-controlled within-subject design and double-blind procedures were used. Subjects were examined twice at a 2 to 4 week interval after either placebo or drug administration (order being counterbalanced). On each test day, subjects underwent baseline testing including psychological and PPI measures. Ninety minutes later, subjects received placebo or MDMA (1.7 mg/kg PO) and were retested after 75 min during the peak of behavioral effects of MDMA. RESULTS: As expected, MDMA decreased prepulse inhibition in a dose-related fashion in rats. In contrast, a typical recreational dose of MDMA (1.7 mg/kg, orally) increased prepulse inhibition in subjects experiencing robust psychological effects. CONCLUSIONS: This surprising disparity between the effects of the drug in rats and humans may reflect a species-specific difference in the mechanism of action of MDMA or in the behavioral expression of a similar pharmacological effect, or both. PMID- 10367554 TI - Residual effect of zolpidem 10 mg and zopiclone 7.5 mg versus flunitrazepam 1 mg and placebo on driving performance and ocular saccades. AB - RATIONALE: Studies report contradictory results concerning the residual effects of zolpidem and zopiclone. Moreover, residual effects of these compounds on healthy subjects have not yet been simultaneously assessed. OBJECTIVE: The present study with healthy subjects investigated the residual effects of zolpidem 10 mg and zopiclone 7.5 mg on driving performance and on ocular saccade and compared them to those under flunitrazepam 1 mg and placebo. METHODS: The study involved 16 subjects divided into two groups, a 9:00 a.m. group and a 11:00 a.m. group, in a balanced, double-blind, cross-over design. RESULTS: In the 9:00 a.m. group, zolpidem had no residual effects while zopiclone and flunitrazepam both impaired driving performance (P < 0.001 for both) and increased saccadic latency (P < 0.005; P = 0.052, respectively). Zopiclone impaired driving performance 5 times less than did flunitrazepam. In the 11:00 a.m. group, zolpidem and zopiclone had no residual effects, while flunitrazepam increased saccadic latency (P = 0.065) but did not impair driving performance. CONCLUSIONS: Zopiclone and flunitrazepam had residual effects in the first part of the morning, whereas zolpidem had no residual effects. The hierarchical character of the effects of the molecules differed according to the test administered. This is probably linked more to drug-induced specific alterations than to different sensitivities of the tests. PMID- 10367555 TI - Relationships of plasma tryptophan availability to course of illness and clinical features of alcoholism: a preliminary study. AB - RATIONALE: Serotonergic (5-HT) mechanisms may be involved in impulse control (including antisocial behavior) across psychiatric syndromes. Age of onset may differentiate alcoholics on psychopathological characteristics associated with impulse control, especially mood disturbance, hostility, and a broad range of antisocial behaviors. Thus, there may be a predictable relationship between markers of 5-HT function and age of onset-related characteristics. OBJECTIVE: We tested the hypothesis that there would be a predictable relationship between the ratio of plasma tryptophan to large neutral amino acids (i.e. TRYP/LNAA ratio), a marker of 5-HT function, age of onset and related psychopathological characteristics associated with impulse control. METHODS: Fifty-eight male and female DSM-IV diagnosed alcoholics attending an outpatient treatment center completing a comprehensive psychopathological assessment, and from whom blood samples were obtained. RESULTS: Plasma TRYP/LNAA ratio was positively correlated with symptoms of dysphoria, and negatively associated with harm avoidance on Cloninger's Temperament and Character Inventory. Low tryptophan availability was associated with antisocial-type personality characteristics. Interestingly, the few (nine) subjects who had both early onset alcoholism and antisocial personality disorder had a higher plasma tryptophan but similar TRYP/LNAA ratio to the others. CONCLUSIONS: These data suggest that a low plasma TRYP/LNAA ratio is associated with susceptibility to anxiety, antisocial-type personality characteristics, and an early age of onset for alcoholism. In contrast, a high plasma TRYP/LNAA ratio is associated with a later onset of alcoholism and dysphoria. PMID- 10367556 TI - Rated well-being in relation to plasma concentrations of l- and d-methadone in satisfied and dissatisfied patients on methadone maintenance treatment. AB - RATIONALE: One of the major problems in methadone maintenance treatment is to find optimal individual doses for the patients. OBJECTIVE: The present study investigated whether the use of rating scales together with enantioselective analysis of l-methadone might facilitate dose adjustments in a clinical situation. METHODS: Rating scales were used to evaluate subjective and objective signs of well-being in relation to plasma methadone concentrations in two groups of patients receiving methadone maintenance treatment. The first group (n = 25) was well-adjusted according to clinical observations and were satisfied with their methadone doses (86.2+/-4.3 mg). The second group (n = 25) was in need of the methadone dose adjustment; they complained of low dosing, despite a dose level of 69.2+/-4.0 mg/day. RESULTS: Results indicated a significant correlation between dose and methadone concentration among dissatisfied patients only. The trough levels of d,l-methadone and l-methadone, as well as their elimination rates, were similar in the two groups of patients. There was a variable predominance of l- over d-methadone in plasma (ratio approximate to 1.2; range 0.7-3.6). Illicit use of drugs by the patients was related to the methadone dose and to satisfaction with the dose received. Increased illicit drug use among dissatisfied patients was successfully eliminated by raising the methadone dose. Subjective and objective ratings of the satisfied patients were quite stable throughout the evaluation period, whereas the ratings of the dissatisfied patients were unstable. These patients seemed to be more sensitive to low trough levels of methadone than the satisfied patients. Associations between the subjective and objective ratings and plasma methadone, along with background characteristics, were characterized by multiple regression analyses. The plasma concentrations of l-methadone were one of the most important explanatory variables in these analyses. Associations between well-being and methadone concentrations in plasma were stronger for l-methadone than for d,l-methadone. CONCLUSIONS: Selective measurements of the active isomer and the use of rating scales should be of clinical value when monitoring methadone maintenance treatment patients. PMID- 10367557 TI - Effect of d-amphetamine on prepulse inhibition of the startle reflex in humans. AB - RATIONALE: Prepulse inhibition of the startle reflex (PPI) is attenuated in animals after administration of d-amphetamine and other drugs that stimulate mesolimbic dopamine activity. OBJECTIVE: The aim of the present study was to evaluate the effects of d-amphetamine (20 mg) on a variety of psychophysiological and subjective measures, including PPI, in humans. METHOD: Thirty-six participants (18 women) participated in a double-blind, placebo controlled, repeated measures study. In one session, participants received d-amphetamine (20 mg) orally, and in the other session, participants received an identical appearing placebo. Participants were assessed at 60, 90, and 120 min after ingestion with a 5-min block of startle trials (six control trials and six prepulse trials) followed by subjective measures of stimulation and mood. RESULTS: d-Amphetamine increased subjective measures of stimulation and euphoria, attenuated PPI, and increased heart rate, relative to placebo treatment. CONCLUSIONS: The effect of d-amphetamine on the subjective measures was substantial and consistent over time, while the effect on PPI was only observed at 90 min after ingestion, and the effect on heart rate was limited to 90 and 120 min after ingestion. PMID- 10367558 TI - In vitro immunoregulatory effects of lithium in healthy volunteers. AB - RATIONALE: There is now some evidence that major depression is associated with activation of the inflammatory response system (IRS). Lithium is effective in the treatment and prophylaxis of major depression and shows significant immunoregulatory functions. OBJECTIVE: The aims of the present study were to examine the in vitro effects of lithium on the unstimulated and lipolysaccharide (LPS) + phytohemagglutinin (PHA)-induced production of proinflammatory cytokines, such as interleukin-6 (IL-6), IL-8, tumor necrosis factor-alpha (TNFalpha) and interferon-gamma (IFNgamma), and negative immunoregulatory cytokines or proteins, such as IL-10 and the IL-1 receptor antagonist (IL-1RA). METHODS: The in vitro effects of lithium carbonate at low (10(-4) M and 10(-5) M) and therapeutic (10( 3) M) concentrations on the above cytokines and the IL-1RA were examined in nine healthy volunteers on whole blood supernatant cultured for 72 h. RESULTS: Lithium (10(-3) M) in the presence of LPS+PHA significantly increased the stimulated production of IFNgamma, IL-8, TNFalpha, IL-1RA and IL-10. Lithium (10(-3) M) significantly increased the unstimulated production of IL-8 and IL-10. CONCLUSIONS: The results suggest that lithium has significant immunoregulatory effects by increasing the production of both proinflammatory cytokines (IFNgamma, TNFalpha and IL-8) and negative immunoregulatory cytokines or proteins (IL-10 and the IL-1RA). PMID- 10367559 TI - Altered open-field behavior in experimental chronic hepatic encephalopathy after single venlafaxine and citalopram challenges. AB - RATIONALE: Latent or manifest chronic hepatic encephalopathy (HE) symptomatology often includes affective symptoms. It is therefore warranted to investigate the functional outcome of novel antidepressants when chronic HE prevails. OBJECTIVE: Portacaval shunt (PCS) in rats is a widely used experimental model for chronic HE, a neuropsychiatric syndrome accompanying liver dysfunction. HE is believed to arise from a primary alteration in neurotransmission in the CNS. PCS has been reported to increase the metabolism of serotonin in the brain, and thus the central serotonin nerve of PCS rats may contain more serotonin than normal. However, the functional relevance of this serotonergic alteration in terms of affecting behavioral performance of PCS rats has been only rarely studied. METHODS: Locomotor and rearing activities were recorded in PCS and sham-operated control rats. A single subcutaneous challenge with saline versus either the mixed serotonin/noradrenaline reuptake inhibitor venlafaxine (10 mg x kg(-1)) or the selective serotonin reuptake inhibitor citalopram (5 mg x kg(-1)) were performed. RESULTS: The PCS-saline injected rats showed reduced locomotor and rearing activity compared with sham-saline treated rats. While no significant differences could be observed following the venlafaxine challenge to controls, the PCS venlafaxine challenged rats displayed reduced behavioral activity as compared to PCS-saline treated rats. The PCS-citalopram rats, however, displayed increased activity compared with the PCS-saline rats while, again, no effect of the citalopram challenge to controls was found. CONCLUSIONS: The present study show altered but opposite behavior in PCS rats, when challenged with either venlafaxine or citalopram, compared to PCS control rats. These findings therefore support the contention that caution should be advocated when CNS monoamine active drugs are used in liver-impaired subjects until better delineation of the combined pharmacodynamic and pharmacokinetic outcome for each such drug in this condition has been made. PMID- 10367560 TI - Cognitive performance in (+/-) 3,4-methylenedioxymethamphetamine (MDMA, "ecstasy") users: a controlled study. AB - RATIONALE: (+/-) 3,4-Methylenedioxymethamphetamine (MDMA, "ecstasy") is an amphetamine analog and drug of abuse. In animals, MDMA damages brain serotonin (5 HT) neurons at doses that overlap with those used recreationally by some humans. To date, few functional sequelae of MDMA-induced 5-HT damage have been identified. OBJECTIVE: Since serotonin is thought to be involved in cognitive processes, and since previous studies have reported verbal and visual memory deficits in MDMA users, the present study sought to determine whether other cognitive processes are influenced by previous exposure to MDMA. METHODS: Twenty two MDMA users who had not used MDMA for at least 3 weeks and 23 control subjects were tested repeatedly with a computerized cognitive performance assessment battery while participating in a 5-day controlled inpatient study. Cerebrospinal fluid (CSF) measures of monoamine metabolites were also collected as an index of brain monoaminergic function. RESULTS: MDMA users and controls were found to perform similarly on several cognitive tasks. However, MDMA subjects had significant performance deficits on a sustained attention task requiring arithmetic calculations, a task requiring complex attention and incidental learning, a task requiring short term memory and a task of semantic recognition and verbal reasoning. MDMA users also had significant selective decreases in CSF 5-HIAA. CONCLUSIONS: The present CSF data provide further evidence that MDMA is neurotoxic to brain 5-HT neurons in humans, and the behavioral data suggest that brain 5-HT injury is associated with subtle, but significant, cognitive deficits. PMID- 10367561 TI - The potentiating effect of sertraline and fluoxetine on amphetamine-induced locomotor activity is not mediated by serotonin. AB - Sertraline dose-dependently increased the locomotor stimulating effect of amphetamine. At the highest dose, 20 mg/kg sertraline had a biphasic effect on amphetamine-induced hyperactivity, producing an initial reduction in amphetamine induced hyperactivity that was later followed by an augmentation of amphetamine induced hyperactivity in the last hour of the 3-h test. Sertraline, at doses of 5 and 10 mg/kg, produced an augmentation of amphetamine-induced hyperactivity over the last 2 h of the 3-h test session. Further, there was an increase in the concentration of amphetamine in the brain in rats pretreated with 5 mg/kg sertraline. Both sertraline (5 mg/kg) and fluoxetine (5 mg/kg) produced an augmentation of amphetamine-induced hyperactivity that was unaltered by a serotonergic lesion of the median and dorsal raphe nuclei that resulted in a greater than 90% depletion of serotonin in hippocampus, striatum, and nucleus accumbens. Further, both sertraline and fluoxetine inhibited spontaneous locomotor activity and this effect was also unaltered by the depletion of serotonin. Thus, serotonergic neurotransmission is not essential for the effects of sertraline and fluoxetine on spontaneous and amphetamine-induced locomotion. It is probable that sertraline and fluoxetine augment the locomotor stimulatory effect of amphetamine by decreasing the metabolism of amphetamine, perhaps via actions on cytochrome P450 isozymes. PMID- 10367562 TI - Resolution of treatment-refractory depression with naltrexone augmentation of paroxetine--a case report. PMID- 10367563 TI - Mechanisms of vascular wall calcium relaxation. AB - Using the method of isometric tension measurement in isolated blood vessels, we investigated some mechanisms of action of high calcium concentrations (>3 mM) on the mechanical activity of small branches of the rat mesenteric artery. Calcium in concentrations up to 30 mM caused relaxation of the arteries (calcium relaxation). The amplitude of the effect decreased in the presence of ouabain (10(-4) M), tetraethylammonium (10(-3) M), charibdotoxin (10(-7) M) and in the potassium-free external solution in intact and denuded rings. Glibenclamide (10( 6) M), 4-aminopyridine (10(-3) M), barium (10(-3) M) and cesium (2.10(-2) M) were inefficient. Calcium relaxation of intact vessels was impaired in the presence of N(omega)-nitro-L-arginine (10(-4) M) or methylene blue (10(-4) M) but not in the presence of indomethacin (10(-5) M). The attenuation of calcium relaxation to the same extent was observed in denuded mesenteric arteries. We conclude that calcium can cause relaxation of vascular smooth muscle cells by two mechanisms. The first is mediated via the cell membrane hyperpolarization due to the activation of Na+/K(+)-ATPase and Ca(2+)-activated potassium channels. The second mechanism is endothelium-mediated and depends on the nitrogen monoxide-guanylate cyclase pathway. PMID- 10367564 TI - Quantitative and ultrastructural analysis of the chondriome in ovogenesis and embryogenesis of the sea urchin Paracentrotus lividus. AB - The dynamics of chondriome in the ovogenesis of the sea urchin Paracentrotus lividus was studied. Growing oocytes 20-30, 50-60 and 90-100 microm in diameter ("small", "medium-sized" and "large", respectively) and mature eggs were used for the ultrastructural and stereological analysis of mitochondria. Linear parameters of mitochondria (length and thickness) were measured on 3-D reconstructions of serial ultrathin sections using the software developed in the laboratory. The following transformations of chondriome structure were shown to occur during ovogenesis: (1) the number of mitochondria (MT) increases with the growth of cytoplasmic compartment; (2) the modal length of MT increases from 0.5 microm in small oocytes to 1 microm in large ones and decreases again to 0.5 microm in the egg; this process is accompanied by changes in the relative number of spherical MT which decreases in medium-sized oocytes and subsequently rises again in the egg; (3) in medium-sized oocytes, dumbbell-shaped MT appear first, the number of these MT reaching the maximum to the stage of large oocytes. In mature eggs, the dumb-bell-shaped MT are absent; (4) in small and medium-sized oocytes, the orthodox conformation of MT is observed, in contrast to MT with a condensed matrix in large oocytes and eggs; (5) in mature eggs, mitochondrial clusters containing 10 to 20 MT of various size are formed. Based on the data obtained, we suggested that during ovogenesis of the sea urchin, specific differentiation of the chondriome is induced which leads to the increase in the quantity of MT via multiple division acts, while restricting the MT growth and variability of their shape. PMID- 10367565 TI - Induction of P-glycoprotein functional activity and multidrug resistance by a soluble factor(s) produced by some mammalian cells. AB - In the previous study we have found that Djungarian hamster fibroblasts with high levels of multidrug resistance (MDR) (colchicine-resistance index RI of 1000 to 42000) produce soluble factor(s) communicating MDR to the drug-sensitive cells of the same species by elevating the functional activity of P-glycoprotein (Pgp). Here we have shown that these cells can influence human tumor cells in the same fashion. Rat hepatoma McA RH7777 cells and their colchicine-resistant derivatives are shown to produce a factor with similar effects (induction of MDR and Pgp functional activity in the drug-sensitive cells). These effects seem to depend on the drug resistance level of the donor cells. Our results show that induction of the Pgp-mediated MDR is not species-specific and the tumor cells with intrinsic MDR (arising from the tissue with a high level of Pgp expression) can produce a factor(s) communicating this type of drug resistance to the sensitive cells. PMID- 10367566 TI - Coordinated regulation of P-glycoprotein activity and cytochrome P-4501A induction in sublines of rat hepatoma McA RH7777 cells with different levels of colchicine resistance. AB - The constitutive and induced activities of cytochrome P-4501A isoforms in hepatoma McA 7777 sublines with different levels of colchicine (CH) resistance were studied. The higher CH resistance was associated with the elevated functional activity of P-glycoprotein (Pgp). The constitutive level of benzo(a)pyrene hydroxylase and 7-ethoxyresorufin O-deethylase (cytochrome P-4501A dependent activities) were the same in sublines with different CH resistance levels. However, benzo(a)-anthracene, a cytochrome P-4501A inducing agent, more effectively induced benzo(a)pyrene hydroxylase and 7-ethoxyresorufin O-deethylase activities in sublines with elevated P-glycoprotein activity. The toxicity of benzo(a)pyrene, a compound which is simultaneously a cytochrome P-4501A-inducing agent and a toxic agent activated by cytochrome P-4501A, is more effective in sublines with elevated CH resistance. These results support the suggestion about the coordinated regulation of enzyme systems involved in the defence against various lipophilic xenobiotics. The possibility to overcome the Pgp-mediated MDR of some tumours by using a combination of some drugs including compounds which induce the cytochrome P-4501A isoforms and are activated by them is discussed. PMID- 10367567 TI - Modulation of the activity of Ca(2+)-activated K+ channels by internal Mg2+ in cultured kidney cells vero. AB - The inside-out mode of the patch-clamp method was used to study the effects of internal Mg2+ on single large-conductance (193+/-7 pS) Ca(2+)-activated K+ channels in cultured kidney cells. In the absence of Ca2+, Mg2+ (1 to 10 mM) did not activate the channels but modified the activating effect of Ca2+ ions: it decreased the Hill coefficient (n), reduced the apparent dissociation constant (K0.5), and modified the channel open and closed times. K0.5 was found to be a voltage-dependent parameter. In the absence of Mg2+, it averaged 600 microM at 20 mV and 27 microM at +30 mV (22 degrees C, pH 6.8). Mg2+ at saturating concentrations (5 to 10 mM) decreased K0.5 to 50 microM at -20 mV and to 15 microM at +30 mV. Irrespective of the membrane potential, K0.5 tended to its limit value of about 12.6 microM. Thus, the effects of membrane depolarization and Mg2+ exhibited a non-additive, competitive relationship. Mg2+ perturbed the exponential shape of the voltage dependences of K0.5. The Hill coefficient characterizing the interaction of Ca2+ ions with the channels was found to be voltage-dependent. In the absence of Mg2+, it increased rather sharply from approx. 2 to 3.5 when the membrane potential was raised from -10 to 0 mV. Mg2+ increased n in a dose-dependent manner; however, about a twofold increase of n occurred within a narrow concentration range (2 to 3 mM). The action of Mg2+ on n was, apparently, voltage-independent, and the effects of Mg2+ and voltage on n were seemingly additive. PMID- 10367568 TI - Effects of surfactants on the activity of phospholipase D. AB - We studied the dependence of the activity of cabbage phospholipase A on the substrate (phosphatidylcholine) the aggregated state of which is regulated by addition of either anionic (sodium dodecyl sulfate, cholate or oleate) or cationic (cetyl-trimethylammonium bromide) surfactants. Activation of the enzyme induced by anionic surfactants was shown to correlate with the size of their polar groups. The phospholipase hydrolase activity correlated with the transformation of multilayer liposomes into micelles. The relationship between the processes was of a complex character. The dependence of the amount of enzymically released choline on the calcium concentration passed through a sharp maximum in the presence of the anionic detergents and monotonically increased in the presence of the cationic detergent. In the former case, the sharp increase in the enzyme activity was suggested to be caused by precipitation of phospholipase D with the anionic detergent calcium salt, which can be considered as a specific type of immobilization. PMID- 10367569 TI - Determination of the fraction composition of blood lipoproteins by the small angle X-ray scattering technique. AB - A new method for analyzing the fraction composition of blood lipoproteins (LP) was developed based on the small-angle X-ray scattering (SAXS) technique. The method allows quantitative determination of the contents of basic LP fractions (high-density LP, low-density LP, very low-density LP and their subfractions) in the blood plasma or serum. The results of LP analysis by the new method were compared with electron microscopy, ultracentrifugation and gel electrophoresis data. The results obtained by SAXS correlated with those obtained by traditional methods. The new method for the determination of the LP fraction composition in the blood is rapid (1-1.5 h), uses only one reagent (e.g., sucrose) and features a high accuracy and resolution up to LP subfractions. A total of 0.05 ml of the blood plasma or serum is required for an assay. The assays can be carried out in purified preparations or in the blood plasma or serum. The method developed can be used in clinical practice for diagnostics and in scientific research. PMID- 10367570 TI - Properties of ionic channels formed by the antibiotic syringomycin E in lipid bilayers: dependence on the electrolyte concentration in the bathing solution. AB - Using the planar lipid bilayer technique, organization of ionic channels formed by the lipodepsipeptide antibiotic syringomycin E applied to one (cis) side of a lipid bilayer was studied. Low concentrations of NaCl (0.01-0.1 M) induced the opening and closing of two types of channels - "small" and "large". The large channels had single channel conductances approximately six times greater than those of the small channels. An increase in the NaCl concentration (0.6-1.0 M) decreased almost completely the chance to reveal the large channels. Although the syringomycin channels exhibited the anion selectivity within the entire range of NaCl concentrations in the bathing solutions (from 0.001 to 1.0 M) whereas the concentration gradients across the bilayers were 2 and 4, the transfer numbers for Cl-decreased with an increase in the mean NaCl concentration (from 0.83 for 0.005 M to 0.70 for 0.5 M). Moreover, at each mean value of NaCl concentration, all conductance levels had the same ion selectivity (identical reversal potential). These results suggest that at low NaCl concentrations the large channels are clusters of small channels which synchronously open and close, while at high electrolyte concentrations the screening of the charged groups responsible for channel interactions prevents the cluster formation. A new theoretical approach for the estimation of the channel radius and the number of elementary charges located at its inner surface (based on the experimental curve of the dependence of transfer number on the NaCl concentration) was developed. Based on this theoretical approach, the channel radius equal to 1 nm and one elementary charge located at its inner surface were obtained. PMID- 10367571 TI - Combined posterior chamber phakic intraocular lens and laser in situ keratomileusis: bioptics for extreme myopia. AB - PURPOSE: To examine the efficacy, predictability, stability, and safety of combined posterior chamber phakic intraocular lens (IOL) implantation and laser in situ keratomileusis (LASIK) in eyes with extreme myopia. METHODS: We analyzed the results of 67 eyes that received a posterior chamber hydrogel-collagen plate phakic IOL (STAAR Collamer Implantable Contact Lens) and also underwent secondary LASIK for the correction of extreme myopia. Mean follow-up was 3 months after the LASIK portion of the procedure (range, 1 day to 6 mo after LASIK). RESULTS: Mean preoperative spherical equivalent refraction was -23.00 +/- 3.60 D (range, -18.75 to -35.00 D), and mean refractive cylinder was 1.50 +/- 1.20 D (range, 0 to 5.00 D). Mean spherical equivalent refraction after IOL implantation and before LASIK was -6.00 +/- 2.80 D (range, -2.00 to -14.38 D) and mean refractive cylinder 1.50 +/- 1.10 D (range, 0 to 5.00 D). Mean postoperative spherical equivalent refraction at last examination after the LASIK portion of the two-part phakic IOL LASIK procedure was -0.20 +/- 0.90 D (range, +1.75 to -5.13 D), and mean refractive cylinder was 0.50 +/- 0.50 (range, 0 to 2.25 D). Eighty-five percent (57 eyes) were within +/- 1.00 D and 67% (45 eyes) were within +/- 0.50 D of emmetropia at last examination. The refractions remained stable with a statistically insignificant change (P > .05 at each interval) during follow-up. Postoperative uncorrected visual acuity at last examination was 20/20 or better in 3% (2 eyes) and 20/40 or better in 69% (46 eyes). A gain of 2 or more lines of spectacle-corrected visual acuity was seen in 51 eyes (76%) and no eyes lost 2 or more lines of spectacle-corrected visual acuity at last examination. CONCLUSION: Combined posterior chamber phakic IOL implantation with the STAAR Collamer plate lens and LASIK (bioptics) is an effective and reasonably predictable method for correcting myopia from -18 to -35 D. Gains in spectacle-corrected visual acuity were common, and results demonstrated good short-term safety and refractive stability. PMID- 10367572 TI - Posterior chamber phakic intraocular lens for hyperopia. AB - PURPOSE: A Phase I U.S. FDA clinical study of a plate haptic posterior chamber phakic intraocular lens (STAAR Surgical Implantable Contact Lens) for treatment of hyperopia was conducted at 4 sites in the United States. The purpose of this report is to assess the short-term safety and efficacy. METHODS: Ten patients with hyperopia between +2.50 and +10.875 D were implanted in one eye each with the posterior chamber plate phakic intraocular lens and were examined at baseline and 1 day, 1 week, 1, 3, and 6 months after surgery. Mean baseline hyperopia was +6.63 D. RESULTS: At 6 months postoperatively, 7 of 10 eyes (70%) had an uncorrected visual acuity of 20/20 or better and 10 of 10 (100%) had 20/40 or better. Eight of ten eyes (80%) had a spectacle-corrected visual acuity within 1 line of baseline; the other two eyes (20%) had an improvement of 3 lines. Mean 6 month postoperative spherical equivalent refraction was +0.20 +/- 0.61D (range, 0.50 to +1.50 D), a reduction of 6.025 D from baseline. Eight of 10 eyes (80%) were within +/-0.50 D of emmetropia, 9 eyes (90%) were within +/-1.00 D, and all eyes (100%) were within +/-1.50 D. No operative or postoperative complications or adverse reactions were observed. CONCLUSIONS: Results support the short-term safety, efficacy, and predictability of the STAAR Surgical Implantable Contact Lens (plate haptic posterior chamber phakic intraocular lens) in the treatment of hyperopia. PMID- 10367574 TI - Topographic predicted corneal acuity with intrastromal corneal ring segments. AB - PURPOSE: To evaluate predicted optical quality of the central anterior corneal surface before and after the intrastromal corneal ring segment (ICRS) refractive procedure using a clinical videokeratoscope and software index developed for that purpose. METHODS: Predicted corneal acuity, a topographically derived index provided with the EyeSys System 2000 videokeratscope, representing potential optical quality of the cornea, was assessed preoperatively and at postoperative month 3 in 94 eyes that received an ICRS to treat -1.00 to -6.00 D of myopia. Predicted corneal acuity was calculated by determining the difference between a measured cornea and its best-fit ellipses for reflected ring circumferences within the central 3 mm diameter zone. RESULTS: Preoperative predicted corneal acuity was 20/10 in 92 of 94 eyes (98%). At month 3 after the ICRS procedure, 48 (51%) of moderately myopic eyes were corrected to 20/20 or better, 96% (90 eyes) were corrected to 20/40 or better, and 98% of eyes (92 eyes) had a predicted corneal acuity of 20/10. For the eyes with a predicted corneal acuity of 20/10, spectacle-corrected visual acuity was normally distributed between 20/10 and 20/25. CONCLUSION: Predicted corneal acuity did not change significantly from baseline in eyes with an ICRS. This suggests that topographic irregularities in the central 3 mm of the cornea detectable by predicted corneal acuity software were not induced in the central cornea with the ICRS. PMID- 10367573 TI - Clear lens extraction with intraocular lens implantation for hyperopia. AB - PURPOSE: Current surgical options for the correction of moderate to severe hyperopia include hyperopic laser in situ keratomileusis (LASIK), phakic intraocular lens implantation and clear lens extraction with intraocular lens (IOL) implantation. We investigate the safety and efficacy of clear lens extraction with IOL implantation to correct hyperopia. METHODS: Phacoemulsification and IOL implantation was performed on 18 eyes of 10 patients. In 16 eyes, the Hoffer-Q formula was used for IOL power calculation and a single IOL was inserted; in the remaining 2 nanophthalmic eyes, the Holladay-II formula was used and two piggy-back IOLs were inserted. RESULTS: Mean preoperative spherical equivalent for distance was +6.17 D (range, +4.25 to +9.62 D). Patients were followed postoperatively for a mean of 10.5 months (range, 4 to 27 mo). Uncorrected visual acuity in all eyes was 20/50 or better with a median uncorrected visual acuity of 20/40 (range, 20/30 to 20/50). Two patients lost 2 lines of spectacle-corrected visual acuity; both of these patients achieved spectacle-corrected visual acuity of 20/30. CONCLUSIONS: Clear lens extraction with IOL implantation is a safe and effective procedure for the correction of moderate to severe hyperopia in the presbyopic age range. PMID- 10367575 TI - Phototherapeutic smoothing as an adjunct to photorefractive keratectomy in porcine corneas. AB - PURPOSE: To study the smoothing effect of phototherapeutic keratectomy (PTK) using masking fluids as an adjunct to standard photorefractive keratectomy ablations. METHODS: Six fresh porcine corneas underwent -6.00, -10.00, and -15.00 D sphere ablations using the VISX Star excimer laser. Multizone treatments to a maximum 6.5 mm radially symmetrical bed were used with a fluence of 160 mJ/cm2. Three of the treatments were supplemented with a thin layer of balanced salt solution and 6 microm of full beam PTK. The corneas were examined by electron microscopy. RESULTS: Smoother treatment zones were apparent in corneas undergoing PTK following PRK. The effect was more marked at higher dioptric ablations. CONCLUSION: PTK may improve surface smoothness after PRK, especially for higher dioptric ablations. PMID- 10367576 TI - Bilateral comparison of photorefractive keratectomy for myopia using two excimer lasers. AB - PURPOSE: The aim of this study was to compare the results of excimer laser photorefractive keratectomy (PRK) in patients who underwent PRK using the Summit Apex (Omnimed) excimer laser in one eye and the Nidek EC-5000 excimer laser in the other. METHODS: All consecutive patients who underwent PRK with the Summit Apex laser (Omnimed) in one eye and the Nidek laser (EC-5000) in the second and had at least 12 months of follow-up were included in this retrospective study (n=30). Uncorrected and spectacle-corrected visual acuity, final spherical equivalent refraction, and grade of subepithelial haze were compared. The average preoperative spherical equivalent refraction of eyes treated with the Summit laser was -6.00 D (range, -2.50 to -8.75 D), and for Nidek-treated eyes it was 5.57 D (range, -2.50 to -8.80 D). RESULTS: Forty-seven percent of Summit-treated eyes and 53% of Nidek-treated eyes had uncorrected visual acuity of 6/6 or better; 61% of Summit-treated eyes and 63% of Nidek-treated eyes had uncorrected visual acuity of 6/7.5 or better; 95% of Summit-treated eyes and 95% of Nidek treated eyes had uncorrected visual acuity of 6/12 or better (difference not statistically significant). Seventy-three percent of eyes treated with the Summit laser and 80% of eyes treated with the Nidek laser had a postoperative refraction within +/-0.50 D of emmetropia; 97% of Summit-treated eyes and 87% of Nidek treated eyes had a postoperative spherical equivalent refraction within +/-1.00 D of emmetropia; the difference between the two lasers was not statistically significant. However, the percent of eyes with persistent hyperopia was smaller in the Nidek group after 3 months (P=.0062) and after 6 months (P=.07) than in the Summit group. Videokeratography was not done. CONCLUSION: Both lasers were effective with relatively low side effects. No significant difference was found between the two lasers in postoperative uncorrected visual acuity or refractive outcome. Eyes operated with the Nidek laser had less persistent hyperopia and stabilized earlier. PMID- 10367577 TI - Correlation of subepithelial haze and refractive regression 1 month after photorefractive keratectomy for myopia. AB - PURPOSE: To relate myopic regression after photorefractive keratectomy (PRK) to subepithelial haze at the first postoperative month. METHODS: One hundred nineteen eyes of 119 patients underwent excimer laser PRK for treatment of myopia up to -8.00 D. Eyes were examined at 1, 3, 6, 9, and 12 months after surgery. All eyes received fluorometholone 0.1% for the first 5 postoperative months in a tapered dose. Dexamethasone 0.1% qid for 1 month was prescribed to all eyes with a spherical equivalent refraction less than plano, followed by an augmented dose of fluorometholone 0.1%. Eyes with myopia greater than -0.75 D at 12 months, as well as those that had received dexamethasone at any postoperative interval- regardless of refractive outcome--were considered to be regressed. Eyes that regressed and those that did not regress were compared statistically (Chi-squared statistical criterion with Yate's correction) regarding haze grade. RESULTS: Forty-seven percent (56 of 119) of eyes regressed. In 89.28% (50 of 56) of eyes, subepithelial haze grade was 1 to 2, and in 10.71% (6 of 56), subepithelial haze was graded 0 to 0.5 at 1 month. Fifty-three percent of eyes (63 of 119) did not regress and in all, subepithelial haze was graded 0 to 0.5 at the first month. The correlation between regression and haze grade 1 or more at the first postoperative month was statistically significant (P<.001). CONCLUSION: Mild to marked subepithelial haze (grade 1 to 2) at the first postoperative month after PRK for myopia is strongly related to regression of initial refractive effect and increasing myopia. PMID- 10367578 TI - Relaxing incision guided by videokeratography for astigmatism after keratoplasty for keratoconus. AB - PURPOSE: To evaluate the efficacy of topographic measurements for relaxing incisions in astigmatism following penetrating keratoplasty for keratoconus. METHODS: Twenty patients (20 eyes) had relaxing incisions between July 1989 and August 1994 for high astigmatism after penetrating keratoplasty for keratoconus. Ten eyes were evaluated using a HaagStreit keratometer (1989-1991) and 10 eyes were evaluated using EyeSys videokeratography (1991-1994). Relaxing incisions were performed at the steep meridians in the donor/host wound. RESULTS: Mean preoperative astigmatism was 7.75 +/- 2.05 D (range, 4.50 to 12.00 D) for the keratometry group, and 6.49 +/- 3.24 D (range, 1.11 to 10.13 D) for the videokeratography group. Mean astigmatism following relaxing incision was 3.90 +/ 2.02 D (range, 0.93 to 6.50 D) for the keratometry group and 3.06 +/- 1.62 D (range, 0.07 to 5.64 D) for the videokeratography group (no significant difference). Vector analysis revealed a vectorial change of 4.64 +/- 2.54 D for the keratometry group and 4.68 +/- 2.08 D for the videokeratography group (no significant difference). Mean spectacle-corrected visual acuity was significantly improved in the topography-guided group following the procedure (P = .021). Complications included perforations in 3 of 20 eyes. Four patients (4 eyes) in the keratometry group and one patient (1 eye) in the videokeratography group had residual astigmatism greater than 5.00 D. Four patients in the keratometry group needed a second procedure of relaxing incision because of irregular (2 eyes) or high (2 eyes) astigmatism. One patient (1 eye) in the videokeratography group needed a second relaxing incision. CONCLUSIONS: Videokeratography as a guide for relaxing incision has some benefits over standard keratometry. Preoperative evaluation with videokeratography did not significantly improve the postoperative astigmatism. Fewer reoperations were needed when videokeratography was used. PMID- 10367579 TI - Endothelial cell surface-associated keratan sulfate after excimer laser photoablation of the anterior rabbit cornea. AB - PURPOSE: The expression of keratan sulfate on the surfaces of corneal endothelial cells is altered when the cells are responding to injury. The purpose of this study was to investigate whether excimer laser surgery affected corneal endothelial cells and the levels of keratan sulfate associated with them. METHODS: We performed 14 bilateral, transepithelial phototherapeutic keratectomies in rabbits using a Nidek EC-5000 excimer laser. Ablations were 6 mm in diameter and 50 microm, 150 microm, or 240 microm deep. At various times following surgery the endothelium was immunolabeled for keratan sulfate and examined by scanning electron microscopy. Four untreated corneas were also examined. RESULTS: Three days after surgery, endothelial cells were not flat but were rounded or domed, a finding that was more pronounced after deeper ablations. No rounded cells, however, were seen at post-operative day 12. Keratan sulfate immunolabel was elevated on endothelial cells 3 days after surgery. By postoperative day 36, its expression was normal under the 50-microm ablations, but remained elevated under one of two 240-microm ablations. CONCLUSIONS: Corneal endothelial cells take on a rounded appearance in the early stages after excimer laser photoablations in rabbits, especially after deeper ablations. The apical surface of the endothelium also transiently expresses elevated levels of cell surface-associated keratan sulfate following surgery. These changes appear to be responses to some aspect of the surgery, and may have physiological implications. PMID- 10367580 TI - Barraquer lecture 1998. Why should refractive surgeons be looking beyond the cornea? PMID- 10367581 TI - Optometry's scope of practice expansion. PMID- 10367582 TI - Epithelial haze, punctate keratopathy, and induced hyperopia after photorefractive keratectomy for myopia. PMID- 10367583 TI - Premature dissemination of technology. PMID- 10367585 TI - Mechanisms of hypoxic coronary vasodilatation in isolated perfused rat hearts. AB - We pharmacologically investigated the potential involvement of nitric oxide (NO), prostacyclin, adenosine, adenosine triphosphate (ATP)-sensitive K (K(ATP)) channel opening and Ca2+-activated K (K(Ca)) channel opening in coronary vasodilatation during 15 min of hypoxia in isolated rat hearts perfused at a constant pressure of 70 mm Hg. The coronary flow suppressed by 10(-4) M Nomega nitro-L-arginine methyl ester (L-NAME), which corresponds to the NO-dependent flow, decreased to almost zero during hypoxia. In contrast, the NO-dependent coronary flow amounted to approximately 40% of the total coronary flow during normoxia. The suppression of coronary flow by 10(-5) M 8-phenyltheophylline (8 PT), which corresponds to the adenosine-dependent flow, was remarkable in the middle and the late phases of a 15-min hypoxia. The coronary flow suppressed by 2 x 10(-6) M glibenclamide, which corresponds to the K(ATP) channel opening dependent flow, depended on the agents added to the perfusate. However, there was a marked increase in coronary flow in the early phase of hypoxia in the heart perfused with the combination of 8-PT, 10(-2) M tetraethylammonium (TEA) and L NAME. During hypoxia, the coronary flow suppressed by TEA, which corresponds mainly to the K(Ca) channel opening-dependent flow, also depended on the agents added to the perfusate. However, during reoxygenation, there was a transient significant increase in any combination of the agents. Our study suggests that hypoxia almost completely inhibits NO production, and that K(ATP) channel opening immediately after hypoxia and subsequent enhanced adenosine production cause a marked hypoxic coronary vasodilatation. It also suggests that K(Ca) channel opening causes vasodilatation during reoxygenation. PMID- 10367584 TI - Neointimal formation after balloon-induced vascular injury in Yucatan minipigs is reduced by oral rapamycin. AB - Rapamycin, a macrolide antibiotic known to prevent allograft rejection, is a potent inhibitor of cell proliferation. Therefore we studied the effects of orally administered rapamycin in a pig model of balloon injury in an attempt to reduce the cellular proliferation and neointimal formation thought to play a role in restenosis. Twenty Yucatan minipigs, divided into groups of 10 animals each, were subjected to balloon inflation of the carotid arteries. One group received the methylcellulose vehicle for rapamycin, whereas the second group was treated for a total of 31 days with 2.0 mg/kg of rapamycin administered daily by oral gavage. This dose and treatment regimen produced significant (p < 0.05) reductions in neointimal area (59%) and in the maximal thickness of the neointima (59%) when comparisons were made with vehicle-treated animals. These effects were accompanied by a significant increase in the lumen area in animals that received rapamycin (33%). Medial area was decreased by 18% in these animals. Blood samples from rapamycin-treated pigs indicated peak concentrations of 1.87 +/- 0.45 and 1.70 +/- 0.24 ng/ml at 2 and 4 weeks after balloon angioplasty, respectively. Significant increases in blood pressure of 21 mm Hg and decreases in heart rate of 25 beats/min also were observed in rapamycin-treated animals relative to those that received vehicle. These results indicate that the antiproliferative effect of rapamycin can be demonstrated after oral dosing in a pig vascular injury model, suggesting a possible therapeutic utility for rapamycin or its analogs in patients undergoing balloon angioplasty. PMID- 10367587 TI - Multiple mechanisms are involved in the acute vasodilatory effect of 17beta estradiol in the isolated perfused rat heart. AB - The purpose of this study was to define the dose-dependent effects of 17beta estradiol on coronary flow and cardiac function in isolated rat hearts and to identify the mechanisms involved in its vasodilator action. Hearts from female and male Wistar rats were perfused at constant pressure (100 mm Hg). Stereoisomer specificity and the mechanism of vasodilation by 17beta-estradiol were examined in female rat hearts. Function was measured by a left ventricular (LV) balloon and coronary flow (CF) with an ultrasonic flowmeter. 17Beta-estradiol at 10(-6), 5 x 10(-6), and 10(-5) M increased CF in female hearts by 5 +/- 2, 27 +/- 4 (p < 0.05 vs. baseline), and 40 +/- 4% (p < 0.05 vs. baseline), respectively. The effect of 17beta-estradiol in hearts from male rats was similar but less pronounced compared with females [deltaCF 8 +/- 3, 19 +/- 3 (p < 0.05 vs. baseline)] and 25 +/- 7% (p < 0.05 vs. baseline; p < 0.05 vs. female 17beta estradiol). Maximum vasodilation by the stereoisomer 17alpha-estradiol was significantly smaller [deltaCF 5 +/- 3, 4 +/- 3 (p < 0.05 vs. female 17beta estradiol) and 14 +/- 1% (p < 0.05 vs. baseline; p < 0.05 vs. female 17beta estradiol)] for 10(-6), 5 x 10(-6), and 10(-5) M. Pretreatment with the NO synthesis inhibitor Nomega-methyl-L-arginine (10(-4) M) had no effect on the maximal vasodilator response to 17beta-estradiol (10(-5) M) [deltaCF 36 +/- 6% (p < 0.05 vs. baseline)]. When hearts were pretreated with the prostaglandin synthesis inhibitor diclofenac (10(-6) M), the maximal vasodilator effect of 17beta-estradiol was partially attenuated [deltaCF 12 +/- 7% (p < 0.05 vs. female 17beta-estradiol)]. Similarly, pretreatment with the K+ATP-blocker glibenclamide (10(-6) M) partially inhibited the maximal vasodilator effect of 17beta-estradiol [deltaCF 22 +/- 6% (p < 0.05 vs. baseline; p < 0.05 vs. female 17beta estradiol)]. Pretreatment with the Ca2+ channel antagonist nifedipine (7.2 x 10( 8) M) completely blocked the vasodilator effect. In isolated perfused rat hearts, 17beta-estradiol induced marked acute coronary vasodilation; this effect is in part gender specific, and in female hearts, largely stereoisomer specific. The dilator effect is mediated predominantly by calcium channel blockade, but prostaglandin release and K+ATP channel activation also are involved. In the isolated perfused rat heart, NO production does not contribute to the acute vasodilator effect of 17beta-estradiol. PMID- 10367586 TI - Interleukin-1alpha stimulated prostacyclin release by increasing gene transcription of prostaglandin H synthase and phospholipase A2 in human vascular endothelial cells. AB - This study was conducted to evaluate the effects of interleukin-1alpha (IL 1alpha) on prostacyclin (PGI2) production in cultured human vascular endothelial cells in association with intracellular Ca2+, inositol 1,4,5-trisphosphate (IP3), and with prostaglandin H synthase (PGHS) and phospholipase A2 (PLA2) gene expression by using the competitive polymerase chain reaction (PCR) method. IL 1alpha did not increase PGI2 production for 15 min, but induced an increase of about three-fold relative to that in controls at 60 and 180 min. IL- 1alpha had no effect on intracellular Ca2+ levels throughout the experimental period. In this study, consistent with previous reports, PGHS-1 messenger RNA (mRNA) was constitutively expressed, whereas PLA2 mRNA was not. After stimulation with IL 1alpha, PLA2 mRNA level showed an eightfold increase within 15 min, and PGHS-2 mRNA level increased by 76-fold within 180 min. PGHS-1 mRNA level was increased 1.6-fold at 180 min. These results suggest the existence of regulatory mechanisms of IL-1alpha-induced PGI2 production, which involve PGHS and PLA2 gene transcription. PMID- 10367588 TI - Heart-rate variability effects of beta-adrenoceptor agonists (xamoterol, prenalterol, and salbutamol) assessed nonlinearly with scatterplots and sequence methods. AB - Full antagonists of the cardiac beta-adrenoceptor improve heart-rate variability (HRV) in humans; however, partial agonism at the beta2-adrenoceptor has been suggested to decrease HRV. We therefore studied the HRV effects of some partial agonists of the beta1- and beta2-adrenoceptors in normal volunteers. Under double blind and randomised conditions (Latin square design), eight healthy volunteers received placebo; xamoterol, 200 mg (beta1-adrenoceptor partial agonist); prenalterol, 50 mg (beta1- and beta2-adrenoceptor partial agonist); salbutamol, 8 mg (beta2-adrenoceptor partial agonist); ICI 118,551, 25 mg (selective beta2 adrenoceptor antagonist); and combinations of each partial agonist with ICI 118,551. Single oral doses of medication (at weekly intervals) were administered at 22:30 h with HRV assessed from the overnight sleeping heart rates. HRV was determined by using standard time-domain summary statistics and two nonlinear methods, the Poincare plot (scatterplot) and cardiac sequence analysis. On placebo, the sleeping heart rate decreased significantly, between 2 and 8 h after dosing. The heart rate with ICI 118,551 was unaltered. Xamoterol, prenalterol, and salbutamol increased the sleeping heart rate. ICI 118,551 blocked the heart rate effects of salbutamol, attenuated those of prenalterol, but did not influence the xamoterol heart rate. The scatterplot (Poincare) area was reduced by beta1-adrenoceptor (xamoterol), beta2-adrenoceptor (salbutamol), and combined beta1- and beta2-adrenoceptor (prenalterol) agonism. A reduction in scatterplot length followed salbutamol, prenalterol alone, and prenalterol in combination with ICI 118,551. The geometric analysis of the scatterplots allowed width assessment (i.e., dispersion) at fixed RR intervals. At higher heart rates (i.e., 25 and 50% of RR scatterplot length), dispersion was decreased after xamoterol, prenalterol, and prenalterol/ICI 118,551. Cardiac sequence analysis (differences between three adjacent beats; deltaRR vs. deltaRRn+1) assessed the short-term patterns of cardiac acceleration and deceleration; four patterns were identified: +/+ (a lengthening sequencing), +/- or -/+ (balanced sequences), and finally -/- (a shortening sequence). Cardiac acceleration or deceleration episodes (i.e., number of times deltaRR and deltaRRn+1 were altered in the same direction) were increased after salbutamol and prenalterol. In conclusion, partial agonism at either the cardiac beta1-adrenoceptor (xamoterol), beta2-adrenoceptor (salbutamol), and beta1- plus beta2-adrenoceptors (prenalterol) altered the autonomic balance toward sympathetic dominance in healthy volunteers; blockade of the beta2-adrenoceptor with the highly selective beta2-antagonist ICI 118,551 prevented the effects of salbutamol on HRV, attenuated the HRV effects of prenalterol, but had no effect on the actions of xamoterol. Agonism at both the beta1- and beta2-adrenoceptor reduced HRV in healthy subjects; the implications for the preventive use of the beta-adrenoceptor compounds in cardiovascular disease warrant further investigation. PMID- 10367589 TI - Levcromakalim decreases cytoplasmic Ca2+ and vascular tone in basilar artery of SAH model dogs. AB - We investigated the effects of levcromakalim, a K+ channel opener, on [Ca2+]i and the contractile force of basilar arteries obtained from normal dogs and subarachnoid hemorrhage (SAH) dogs. The responsiveness to serotonin was increased more in the SAH group than in the control group. Levcromakalim decreased the resting [Ca2+]i and force more profoundly than did a Ca2+ channel blocker, nicardipine, and these effects were more prominent in the SAH group than in the control group. Levcromakalim diminished the increases in [Ca2+]i and contractile force induced by serotonin more profoundly than nicardipine did, and these effects were equal in both groups. The effects of levcromakalim were not inhibited by iberiotoxin but were antagonized completely by glibenclamide. These results suggest that levcromakalim-induced opening of adenosine triphosphate (ATP)-sensitive K+ (K(ATP)) channels reduces [Ca2+]i more effectively than does nicardipine and that levcromakalim exerts the vasodilator effects under the condition in which large conductance Ca2+-activated K+ (BK) channels are blocked with iberiotoxin. Consequently, K+ channel openers like levcromakalim may be useful drug candidates to treat delayed cerebral vasospasm after SAH. PMID- 10367590 TI - Central effects of endothelin and its antagonists on sympathetic and cardiovascular regulation in SHR-SP. AB - Intracerebroventricular injections of endothelin-1 (ET-1) are reported to cause dose-related increases in sympathetic nerve activity and blood pressure in anesthetized normotensive rats. These studies were performed to determine the following: which endothelin receptor, A or B, is involved in mediating sympathetic and cardiovascular effects of ET-1 injected centrally; whether central endothelin tonically participates in blood pressure regulation in normotensive rats; and whether the altered endothelin system in the central nervous system contributes to blood pressure elevation in hypertensive rats. ET 1, ET-A antagonist (BQ-123), or ET-B antagonist (RES-701-1) was injected into the lateral cerebral ventricle (i.c.v.) of urethane-anesthetized normotensive Wistar and Wistar-Kyoto (WKY) rats, spontaneously hypertensive rats (SHRs), and stroke prone SHRs (SHR-SPs). In Wistar rats, i.c.v. injections of ET-1 (1, 5, 10 pmol) consistently increased sympathetic nerve activity, thereby elevating blood pressure in a dose-related manner. The pressor responses induced by i.c.v. ET-1 were abolished after intravenous pretreatment with phentolamine. Neither ET-A nor ET-B antagonist, when injected centrally, altered basal levels of sympathetic nerve activity, heart rate, or blood pressure in Wistar rats. However, sympathetic activation and pressor responses induced by i.c.v. injection of endothelin were completely abolished after i.c.v. pretreatment with ET-A antagonist but were unaffected after pretreatment with ET-B antagonist. Although i.c.v. injections of ET-1 increased sympathetic nerve activity and blood pressure in WKY rats, SHRs, and SHR-SPs, the magnitudes of these responses did not differ among these three groups. In contrast, i.c.v. injections of ET-A antagonist decreased sympathetic nerve activity, blood pressure, and heart rate only in SHR SPs, but not in WKY rats and SHRs. In addition, the depressor effects of i.c.v. ET-A antagonist in SHR-SPs were ascertained while these rats were awake. In summary, i.c.v. injections of ET-1 increased sympathetic nerve activity and blood pressure via ET-A receptors but not via ET-B receptors. Central ET might tonically activate sympathetic nerve activity to thereby contribute to blood pressure elevation in SHR-SPs, but not in WKY rats and SHRs. PMID- 10367592 TI - Effect of calcium channel blocker amlodipine on the intimal-medial thickness of carotid arterial wall in type 2 diabetes. AB - Although several reports have suggested that calcium channel blockers may inhibit progression of atherosclerosis in animals, it is still controversial whether they have any clinically significant antiatherogenic action in humans. The measurement of intimal-medial thickness (IMT) of the common carotid artery by B-mode ultrasound technique has been recognized as a powerful and noninvasive method to evaluate early atherosclerotic lesions. We investigated the effect of treatment with amlodipine, a powerful calcium channel blocker, on IMT. Twenty-two hypertensive patients with type 2 diabetes were enrolled in a prospective open study. An amlodipine group (amlodipine, 5 mg; n = 11) and a control group receiving angiotensin-converting enzyme inhibitors (n = 11) were studied before and 6 months after treatment. Amlodipine treatment caused a significant decrease in IMT compared with control (-0.052 +/- 0.017 vs. 0.011 +/- 0.021 mm; p < 0.05). Although the exact mechanisms remain to be elucidated, our preliminary result suggests that amlodipine has an antiatherogenic action in type 2 diabetes. PMID- 10367591 TI - Role of A2A receptor in the modulation of myocardial reperfusion damage. AB - Adenosine protects myocardium from ischemia and reperfusion damage; however, the mechanism of action is still under discussion. We investigated whether (a) adenosine protects isolated crystalloid-perfused rabbit heart from ischemia/ reperfusion injury; (b) this action is receptor mediated and what receptor subtypes are involved, and (c) this action is dependent on an enhanced nitric oxide production. Our results showed a cardioprotective effect of adenosine (10( 4) M), of nonselective adenosine-receptor agonist 5'-N-ethyl-carboxamidoadenosine (NECA; 5 x 10(-6) M), and of A2A agonists CGS 21680 (10(-8) and 10(-6) M), 2 hexynylNECA (10(-7) M). On the contrary, A1 agonist CCPA (10(-8) and 10(-6) M) does not provide any protection. The effect has been achieved in terms of significant reduction in contracture development during reperfusion [diastolic pressure was 46.8 +/- 7.1 mm Hg (p < 0.01); 46.1 +/- 7.8 mm Hg (p < 0.01); 46.9 +/- 5.5 mm Hg (p < 0.01); and 59.3 +/- 6.7 mm Hg (p < 0.05) with 10(-4) M adenosine, 5 x 10(-6) M NECA, 10(-6) M CGS 21680, and 10(-7) M 2-hexynylNECA, respectively, versus 77.6 +/- 5.0 mm Hg in control]; reduced creatine phosphokinase release (13.5 +/- 1.6, 22.2 +/- 7.9, 14.2 +/- 3.3, and 14.1 +/- 4.5 U/gww in treated hearts vs. 34.6 +/- 7.2 U/gww in controls; p < 0.05); improved energy metabolism [adenosine triphosphate (ATP) content is 9.9 +/- 0.5, 10.4 +/- 0.6, 9.8 +/- 0.5, and 10.5 +/- 0.5 micromol/gdw in treated hearts vs. 7.6 +/- 0.2 micromol/gdw; p < 0.05]. Moreover, our data indirectly show a functional presence of A2A receptors on cardiomyocytes as the protection is A2A mediated and exerted only during reperfusion, although in the absence of blood and coronary flow changes. These activities appear independent of nitric oxide pathways, as adenosine and 2-hexynylNECA effects are not affected by the presence of a nitric oxide-synthase inhibitor (10(-4) M L-NNA). PMID- 10367593 TI - Interspecies comparison of the antiplatelet, antithrombotic, and hemorrhagic effects of SR 121566A, a novel nonpeptide GP IIb/IIIa antagonist. AB - We report the results from an interspecies comparison of the antiplatelet, antithrombotic, and hemorrhagic actions of SR 121566A, a novel nonpeptide antiplatelet agent with high affinity and specificity for the GP IIb/IIIa complex. SR 121566A exhibited in vitro antiplatelet activity against adenosine diphosphate (ADP)-induced aggregation with a rank order of potency [humans = baboons = dogs] > marmosets > guinea pigs > rabbits. These in vitro findings were predictive for the ex vivo antiplatelet potency after i.v. administration of SR 121566A to dogs, guinea pigs, and rabbits [median effective dose (ED50) values, 0.02, 0.05, and 0.15 mg/kg]. The antiplatelet actions of SR 121566A translated into an acute antithrombotic effect in an arteriovenous shunt model after i.v. administration in dogs, guinea pigs, rabbits, and marmosets (ED50, 0.08, 0.10, 0.50, and 0.007 mg/kg). Hemorrhagic effects of SR 121566A were observed in guinea pigs and rabbits at doses that represented 2-3 times the antithrombotic ED50, whereas in marmosets, no bleeding was observed at the antithrombotic ED90. These results demonstrate that SR 121566A exhibits favorable actions in terms of antithrombotic potency and hemostatic safety in different animal species, suggesting that, in humans, SR 121566A will be a good candidate as an antithrombotic compound. PMID- 10367594 TI - Inhibition by fendiline of the transient outward current in rat ventricular cardiomyocytes. AB - The effects of fendiline on the transient outward current (Ito) were investigated in rat ventricular cardiomyocytes. Extracellularly applied fendiline reduced peak and steady-state current amplitude of Ito; the inactivation of Ito was accelerated by the drug, which reflects onset of block. The described effects were concentration dependent: half-maximal effects were achieved at approximately 3 microM fendiline. Intracellularly applied fendiline (3 microM) did not affect Ito within 5 min. The steady-state current amplitude of Ito was more efficiently suppressed by the drug at 22 +/- 1 degrees C than at 36 +/- 1 degrees C. The recovery of Ito was analyzed by the application of twin depolarizing voltage pulses, interrupted by variable pulse intervals. In the presence of fendiline, recovery of Ito was about twofold slower than that under control conditions, independent of the drug concentration used, which reflects offset from block. Concentration-dependent onset but concentration-independent offset of block suggest that the described time constants correspond to voltage-dependent net binding and unbinding, respectively, of fendiline at its receptor sites. It is proposed that fendiline binds extracellularly at positive potentials to Ito channels in their open state and dissociates from the channels at rest. PMID- 10367595 TI - Comparison of anti-M2-muscarinic effect of AF-DX 116 on atrioventricular nodal conduction with those of pirenzepine and atropine as antibradyarrhythmic drugs. AB - Selectivity of antimuscarinic actions of AF-DX 116 (AF-DX) on the atrioventricular (AV) nodal conduction was compared with those of pirenzepine and atropine by using the canine isolated, blood-perfused AV node preparation and the open-chest in situ dog heart. In the isolated AV node preparation, dose-response curves for negative dromotropic effects (prolongation of Atrio-His interval) of carbachol (CCh) injected into the posterior septal artery were shifted to the right in parallel by AF-DX, pirenzepine, and atropine with apparent pA2-values of 13, 27.5, and 0.45 microg, respectively, and slopes of the modified Schild plot of nearly unity. Meanwhile, dose-response curves for coronary vasodilator effects of CCh were shifted to the right by AF-DX, pirenzepine, and atropine with the apparent pA2 values of 68, 12.5, and 0.55 microg, respectively, but the slopes were far from unity. In the in situ open-chest heart, dose-response curves for negative dromotropic effects (prolongation of AV conduction time) of CCh given intravenously were shifted to the right in parallel by AF-DX, pirenzepine, and atropine with apparent pA2 values of 36, 32, and 1.25 microg/kg, respectively, and the slope of nearly unity, whereas dose-response curves for hypotensive effects of CCh were shifted to the right by AF-DX, pirenzepine, and atropine with apparent pA2 values of 105, 15, and 0.65 microg/kg, respectively, but the slopes of AF-DX and pirenzepine were far from unity. In addition, prolongations of AV conduction time by electrical stimulation of the left vagus nerve in the in situ heart were suppressed by AF-DX, pirenzepine, and atropine with the ID50, dose for 50% suppression, of 40, 35, and 1.9 microg/kg, respectively. These results suggest that (a) the potency of antimuscarinic actions of AF-DX on the CCh induced negative dromotropic effects was almost equal to that of pirenzepine, and approximately 30 times less potent than atropine; (b) the M2-subtype selectivity of AF-DX was considerably higher in comparison with pirenzepine and atropine; and (c) the muscarinic receptor subtype on the canine AV node is entirely of the M2 type, but only sparsely developed in the coronary vascular beds. PMID- 10367597 TI - Effects of Bay Y5959 on Ca2+ currents and intracellular Ca2+ in cells that have survived in the epicardial border of the infarcted canine heart. AB - We determined and compared the effects of the dihydropyridine agonist, Bay Y5959, on the amplitude of L-type Ca2+ currents and intracellular Ca2+ transients in epicardial cells from noninfarcted hearts (NZs) and surviving cells from the epicardial border zone of 5-day infarcted canine hearts (IZs). We determined the effects of Bay Y5959 on the L-type Ca2+ current by using single cells and a whole cell voltage-clamp approach. To elucidate the effects of Bay Y5959 on the amplitude and time course of the spatially averaged intracellular Ca2+ transient (Ca(i)T), myocytes from the two cell groups were loaded and studied by using the Ca2+-sensitive indicator fura-2/AM. Bay Y5959 increased the amplitude of the L type Ca2+ current in both cell groups, but peak amplitude in NZs was always greater than that in IZs. Bay Y5959 also increased Ca(i)T amplitude in both NZs and IZs and significantly accelerated the Ca(i)T time course in IZs, particularly at the faster pacing-cycle length. We suggest that the Bay Y5959 effect to restore L-type Ca2+ currents in IZs contributes to its observed antiarrhythmic effects during the reentrant ventricular tachycardias that are known to occur in the epicardial border zone of the infarcted heart. PMID- 10367596 TI - Renal effects of L-DOPA in heart failure. AB - We examined whether low-dose L-DOPA treatment induces natriuresis and diuresis in patients with congestive heart failure who have cardiac decompensation despite treatment with digoxin, a diuretic, and an angiotensin-converting enzyme inhibitor and who respond acutely to intravenously infused dopamine. In a randomized, double-blind, placebo-controlled crossover study, 11 patients with severe congestive heart failure received L-DOPA (0.10 g, p.o., t.i.d., for 1 day and then 0.25 g, p.o., t.i.d., for 2 days after a washout period of > or = 1 day), with assessments of plasma and urinary levels of catechols, urinary volume, and sodium content, and clinical and laboratory measures of improvement of congestive heart failure. L-DOPA elicited short-term, dose-related increases in urinary volume and sodium excretion. At the 0.10-g dose, L-DOPA increased plasma L-DOPA levels and urinary L-DOPA excretion by about fivefold, whereas at the 0.25 g dose, L-DOPA increased plasma and urinary L-DOPA by >50-fold. Twenty-four-hour urinary dopamine excretion increased by about fivefold after the low dose of L DOPA and approximately 50-fold after the high dose. The results demonstrate that oral L-DOPA treatment can produce beneficial natriuretic and diuretic effects in selected patients with congestive heart failure. The bioavailability of oral L DOPA appears to vary with the dose. These results support findings from previous studies about beneficial cardiac functional effects of L-DOPA in patients with refractory heart failure. PMID- 10367598 TI - Nitric oxide decreases microvascular permeability in bradykinin stimulated and nonstimulated conditions. AB - This study examined the occurrence of endothelial nitric oxide (NO)-synthase (NOS III) in terminal mesenteric vessels and the involvement of NO in microvascular permeability. Possible effects were studied in bradykinin (BK)-induced and basal conditions. NOS expression was investigated by using NOS-III immunohistochemistry and nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase histochemistry on the light- and electron-microscopic levels. Permeability was examined in dissected mesenteries of male rats weighing 250-300 g. Tissue treatment was performed with BK (100 nM), sodium nitroprusside (SNP, 1 and 10 microM), L nitroarginine (L-NA, 300 microM), BK and L-NA, BK and SNP, L-NA and SNP, as well as with BK, SNP (10 microM), and the guanylylcyclase inhibitor ODQ (10 microM), and BK and ODQ alone. Pharmacologically induced permeability changes were studied with fluorescein isothiocyanate (FITC)-dextran 70 kDa as a tracer for macromolecular transport. Video images were analyzed with computer determination of integrated optical density (IOI). Results were statistically verified by analysis of variance and t test. Microvascular permeability was increased by 168% after BK treatment and was enhanced by NO-synthesis inhibition with L-NA by 607%. However, the NO donor SNP led to a reduced tracer extravasation to 105 and 58%, respectively, an effect blocked by ODQ. Under basal conditions without prior BK induction, L-NA also causes an increase of IOI by 25%, whereas coapplication with SNP resulted in only a 10% increase of permeability. These results point out that NO has a modulatory role for microvascular permeability by supporting the barrier function of the endothelial lining in stimulated and nonstimulated conditions. PMID- 10367599 TI - Long-term administration of atrial natriuretic peptide in patients with acute heart failure. AB - A short-term treatment of atrial natriuretic peptide (ANP), a circulating hormone of cardiac origin, is reported to improve cardiac performance in patients with chronic heart failure. However, clinical usefulness of long-term administration of ANP in patients with congestive heart failure has not been reported. We studied 36 patients with severe acute heart failure who resisted various therapy. Hemodynamic parameters were measured before and 48 h after initiating ANP infusion (n = 18) or normal saline (n = 18). Mean pulmonary capillary wedge pressure (23-->13 mm Hg), mean right atrial pressure (10-->5 mm Hg), systemic vascular resistance (2,169-->1,307 dyn x s x cm(-5)) and pulmonary vascular resistance (318-->136 dyn x s x cm(-5)) decreased significantly, whereas cardiac index (1.9-->2.6 L/min/m2) and urine volume (1,692-->2,560 ml/day) increased during long-term ANP infusion (before-->48 h). Moreover, in eight patients with long-term ANP infusion, these hemodynamic effects were maintained at 7 days after initiating ANP infusion. Vasodilating, pulmonary vasorelaxant, and diuretic activities of ANP are maintained without tolerance, and thus long-term ANP infusion is clinically useful in patients with severe acute heart failure. PMID- 10367600 TI - Blood pressure response to the first 36 hours of heart failure therapy with perindopril versus captopril. French General Hospitals National College of Cardiologists. AB - An open randomized hospital study conducted in 169 centers in France compared the blood pressure response to the first 36 h of treatment with perindopril (PER), 2 mg once daily, with that to captopril (CAP), 6.25 mg t.i.d., in 725 patients (mean age, 70 years; men, 67%) with echocardiographic left ventricular systolic dysfunction (fractional shortening, < or = 28%) due to ischemia (56.7%) or hypertension (34.5%) and a systolic blood pressure (SBP) > or = 120 mm Hg. Each dose of CAP induced a sharp and rapid decrease in blood pressure (maximum, 1.5-2 h); with PER, the decrease was gradual (maximum, 6 h) and variation was less marked. However, at 36 h, the decrease in blood pressure versus baseline was similar on both treatments. Over the 36-h period, there were 22 (3%) dropouts due to marked orthostatic hypotension (SBP, <90 mm Hg), and they were fewer with PER than with CAP: 16 cases in the CAP group versus six in the PER group (p = 0.036). At 36 h, heart rate was lower with CAP than with PER: 75.2 versus 77.5 beats/min, respectively (p = 0.039). As initial therapy for stabilized left ventricular systolic dysfunction, the first dose of PER (2 mg) induced a significantly smaller decrease in blood pressure than the first dose of CAP (6.25 mg); dropouts due to orthostatic hypotension were also significantly fewer with PER than with CAP. PMID- 10367601 TI - Inhibitory mechanisms by which suramin may attenuate neointimal formation after balloon angioplasty. AB - Restenotic neointimal lesions, a major limitation to coronary angioplasty, develop in response to diverse signals and depend on three properties of activated arterial smooth muscle cells (SMCs): proliferation, migration, and abnormal production of extracellular matrix. Most of the pharmacologic approaches targeting specific pathogenic factors facilitating development of restenosis have failed in clinical trials. Our results indicate that the polysulfonated naphthylurea suramin, a "non-specific drug" that interferes with multiple cellular proteins, inhibits neointimal formation in rabbit iliac arteries after balloon-catheter injury administered throughout the critical period of several weeks after the procedure. In vitro studies aimed at dissecting the mechanism(s) underlying the suramin-dependent effect demonstrated that, in addition to an inhibitory effect on SMC proliferation, suramin inhibited fibronectin and elastin deposition and the migration of SMCs through elastin membranes and into scratch gaps of monolayer cultures. We also demonstrated that suramin causes cell-surface accumulation of the elastin binding protein, a receptor that not only anchors SMCs to the extracellular matrix, but also inhibits SMC response to interleukin 1beta (IL-1beta). We conclude that suramin acts as a multitarget inhibitor of SMC activation and has a therapeutic potential as an agent that may attenuate arterial restenosis after angioplasty. PMID- 10367602 TI - Praise be to psychosomatic medicine. AB - OBJECTIVE: To celebrate the 60th anniversary of the publication of Psychosomatic Medicine, a review of its first 45 years of existence was performed to highlight the conceptual, methodological, and scientific advances made. METHOD: A very selective overview of key articles was made to illustrate the evolution of concepts and to document scientific progress. CONCLUSION: Psychosomatic Medicine has published important contributions to the development of an integrative theory of medicine. PMID- 10367603 TI - Reduction of natural killer cytotoxic activity in major depression: interaction between depression and cigarette smoking. AB - OBJECTIVE: Epidemiological data suggest that the presence of a depressed mood combined with cigarette smoking increases the risk of cancer at sites associated with smoking and at sites not associated with smoking. This study tested the hypothesis that major depression and smoking together contribute to a decline of natural killer cell (NK) activity, an immune parameter thought to be important in immune surveillance. METHODS: A sample of 245 men were stratified into four groups: control subjects who were not smokers, control subjects who were smokers, subjects with major depression who were not smokers, and subjects with major depression who were smokers. Blood samples were obtained for measurement of total white blood cell (WBC) counts, differential cell counts, and assay of NK activity. RESULTS: Major depression and cigarette smoking interact and were together associated with changes in WBC counts and NK activity. Depressed subjects who were smokers had higher WBC counts (p < .001) and lower NK activity (p < .01) than depressed nonsmoking subjects. However, WBC counts and NK activity were similar in control smokers and nonsmokers. Backward elimination regression analyses showed that the interaction of depression and smoking significantly (p < .001) predicted WBC counts and NK activity. CONCLUSIONS: This study extends previous findings of immune alterations in patients with major depression. Major depression and smoking interact and together contribute to an elevation of total WBC count and a decline of NK activity. PMID- 10367604 TI - Psychoneuroimmunology and immunotoxicology: implications for carcinogenesis. PMID- 10367605 TI - Relations of trait depression and anxiety to low lipid and lipoprotein concentrations in healthy young adult women. AB - OBJECTIVE: Recent evidence suggests that naturally occurring low cholesterol concentrations (<4.14 mmol/liter) are associated with depression as well as poor psychological health. For the most part, these associations have been observed in men. The current study assessed the relation of naturally occurring low lipid and lipoprotein concentrations to trait measures of depression and anxiety in 121 healthy young adult women. METHODS: Fasting lipid samples were collected at the same time as health history. Trait depression and anxiety were assessed using the Neuroticism, Extraversion, Openness-Personality Inventory (NEO-PI) depression subscale and Spielberger's Trait Personality Inventory (STPI) anxiety subscale. Analyses were conducted using both univariate and multivariate procedures. RESULTS: NEO depression was inversely associated with total cholesterol (p = .027), triglycerides (p = .012), and the ratio of total cholesterol to high density lipoprotein cholesterol (p = .059). Similarly, STPI anxiety was inversely associated with total cholesterol (p = .002), low-density lipoprotein cholesterol (p = .016), triglycerides (p = .024), and ratio of total cholesterol to high density lipoprotein cholesterol (p = .075). These associations were significant after adjustment for age, body mass index, physical activity, oral contraceptive use, and hostility. Neither depression nor anxiety was associated with high density lipoprotein cholesterol. Univariate analyses indicated that women with low total cholesterol concentrations (<4.14 mmol/liter), relative to those with moderate to high cholesterol levels, were more likely to have higher scores on the NEO depression subscale (27 of 69 (39%) vs. 10 of 52 (19%)) and STPI anxiety subscale (24 of 69 (35%) vs. 11 of 52 (21%)). CONCLUSIONS: In healthy young adult women, low lipid and lipoprotein concentrations are inversely associated with trait measures of depression and anxiety. These findings are independent of age, body mass index, physical activity, and other factors known to influence lipid concentrations. PMID- 10367607 TI - Cardiovascular and endocrine changes during sexual arousal and orgasm. PMID- 10367606 TI - Cardiovascular and endocrine alterations after masturbation-induced orgasm in women. AB - OBJECTIVE: The present study investigated the cardiovascular, genital, and endocrine changes in women after masturbation-induced orgasm because the neuroendocrine response to sexual arousal in humans is equivocal. METHODS: Healthy women (N = 10) completed an experimental session, in which a documentary film was observed for 20 minutes, followed by a pornographic film for 20 minutes, and another documentary for an additional 20 minutes. Subjects also participated in a control session, in which participants watched a documentary film for 60 minutes. After subjects had watched the pornographic film for 10 minutes in the experimental session, they were asked to masturbate until orgasm. Cardiovascular (heart rate and blood pressure) and genital (vaginal pulse amplitude) parameters were monitored continuously throughout testing. Furthermore, blood was drawn continuously for analysis of plasma concentrations of adrenaline, noradrenaline, cortisol, prolactin, luteinizing hormone (LH), beta-endorphin, follicle stimulating hormone (FSH), testosterone, progesterone, and estradiol. RESULTS: Orgasm induced elevations in cardiovascular parameters and levels of plasma adrenaline and noradrenaline. Plasma prolactin substantially increased after orgasm, remained elevated over the remainder of the session, and was still raised 60 minutes after sexual arousal. In addition, sexual arousal also produced small increases in plasma LH and testosterone concentrations. In contrast, plasma concentrations of cortisol, FSH, beta-endorphin, progesterone, and estradiol were unaffected by orgasm. CONCLUSIONS: Sexual arousal and orgasm produce a distinct pattern of neuroendocrine alterations in women, primarily inducing a long-lasting elevation in plasma prolactin concentrations. These results concur with those observed in men, suggesting that prolactin is an endocrine marker of sexual arousal and orgasm. PMID- 10367608 TI - Testosterone, gonadotropin, and cortisol secretion in male patients with major depression. AB - OBJECTIVE: Previous studies of sex hormone concentrations in depression yielded inconsistent results. However, the activation of the hypothalamic-pituitary adrenal system seen in depression may negatively affect gonadal function at every level of regulation. The objective of this study was to explore whether major depressive episodes are indeed associated with an alteration of gonadal function. METHODS: Testosterone, pulsatile LH secretion, FSH, and cortisol were assessed using frequent sampling during a 24-hour period in 15 male inpatients with major depression of moderate to high severity and in 22 healthy comparison subjects (age range 22-85 years). RESULTS: An analysis of covariance model showed that after adjustment for age only, daytime testosterone (p < .01), nighttime testosterone (p < .05), and 24-hour mean testosterone secretion (p < .01) were significantly lower in the depressed male inpatients. There was also a trend for a decreased LH pulse frequency in the depressed patients (p < .08). CONCLUSIONS: Gonadal function may be disturbed in men with a depressive episode of moderate to high severity. PMID- 10367609 TI - Effect of autonomic nervous system manipulations on gastric myoelectrical activity and emotional responses in healthy human subjects. AB - OBJECTIVE: The aim of this study was to determine the gastric myoelectrical and emotional responses provoked by two psychophysiological stimuli known to cause in one case increased sympathetic nervous system activity and in the other increased parasympathetic nervous system activity. METHODS: Electrogastrograms (EGGs) were recorded, and interbeat intervals (IBIs) were obtained from electrocardiographic recordings from 20 subjects during baseline and in response to a shock avoidance task (shock stimulus) and forehead cooling (dive stimulus). After each experimental period, subjects reported their emotional experience by rating descriptors ranging from serenity to excitement. RESULTS: During the shock stimulus, IBIs decreased significantly (p < .05), gastric tachyarrhythmias increased (p < .05), and emotional arousal increased, as indexed by reports of increased interest, excitement, and activation. In contrast, during the dive stimulus, IBIs increased (p < .05), but there were no associated changes in gastric myoelectrical activity or emotional arousal. CONCLUSIONS: Acute stress can evoke arousal and dysrhythmic gastric myoelectrical activity, and these acute changes, which occur in healthy individuals, may provide insight into functional gastrointestinal disorders. PMID- 10367610 TI - Critical life events, infections, and symptoms during the year preceding chronic fatigue syndrome (CFS): an examination of CFS patients and subjects with a nonspecific life crisis. AB - OBJECTIVE: The purpose of this study was to describe the sequence of psychosocial events and infections preceding the onset of chronic fatigue syndrome (CFS). This information was related to the temporal development of crucial symptoms in relation to the onset of, namely, fatigue, sadness, irritability, pain, and feeling of fever. METHODS: A personal interview was conducted in 46 patients (mean age, 39.5 years; SD, 9 years) who fulfilled international CFS criteria. These patients were matched with regard to age and gender to 46 carefully matched control subjects. Twenty-three percent of the study subjects were men, and 77% were women. The patient at first identified the month that coincided with the onset of CFS. Similarly, each control subject was asked to identify a "very difficult period" within approximately the same period as the patient with whom the control subject was matched. A list of 14 different life events was perused. Participants were asked to identify for each month whether each of the listed events had occurred. Furthermore, they were asked to rate the importance of the events they had experienced. In addition, for each of the cardinal symptoms (fatigue, sadness, irritability, pain, and feeling of fever) and for each month, the subjects were asked to rate, on a visual analogue scale, the symptom intensity. Also, the number of infections was noted. RESULTS: A statistically significant group difference in fatigue intensity existed during the period 4 to 10 months before the onset of CFS. During the 3 months preceding the diagnosis for the CFS patients or the peak of the crisis for the control group, there was a dramatic rise in fatigue in both groups. The CFS group reached a much higher fatigue level, which leveled off somewhat during the first year of follow-up but still remained very high in comparison with the control group, which reached precrisis levels 4 months after the peak. Similar patterns were observed for fever and pain. With regard to sadness and irritability, no group difference was observed during the period preceding the crisis. In the patient group, the level stayed high throughout the whole first year of follow-up, whereas a slow return started in the control group; precrisis levels were reached after 1 year in this group. The prevalence ratio (CFS patients/control subjects) for negative events was around 1.0 for the periods 4 to 12 months preceding CFS but 1.9 during the quarter year preceding the onset. For infections, the prevalence ratio increased successively during the four quarters preceding CFS (from 1.4 to 2.3). CONCLUSIONS: According to the retrospective self-reports, there were differences between the groups in fatigue, pain, and feeling of fever during the months preceding the crisis. With regard to depressive and irritable feelings, no preillness differences were reported between the groups. There was a reported excess prevalence of both infections and negative life events during the quarter year preceding the onset of CFS or crisis. Potential sources of error are discussed. These findings must be replicated in longitudinal studies. PMID- 10367611 TI - Relationship of physical symptoms and mood to perceived and actual blood pressure in hypertensive men: a repeated-measures design. AB - OBJECTIVE: Noncompliance with antihypertensive treatment is a significant health concern. Researchers have suggested that the absence of definable symptoms associated with elevated blood pressure (BP) attenuates patients' motivation to use medication. The current study evaluated the relation of psychological variables, including symptoms, perceptions of BP, and perceptions of medication efficacy, to physiological variables, including actual BP and the use of active antihypertensive medication vs. placebo. METHODS: Participants included 54 mildly hypertensive men who were participating in a placebo-controlled, double-blind study of the quality-of-life effects of antihypertensive therapies. Survey data and BP measurements were obtained during a series of clinic visits. RESULTS: Mixed-model analysis of variance was used to evaluate both between- and within person relations of psychological to physiological state. Results revealed significant within-person associations between predicted and actual BP. Negative mood was closely related to predicted, but not actual, BP. Participants were also relatively accurate in rating active medications as more effective than placebo. Between-persons analyses did not show relations of symptoms or moods to actual BP. CONCLUSIONS: The significant within-person relations of estimated to actual BP suggest that some individuals may be able to estimate their own BP, although the accuracy of these estimates is limited. The findings may explain patients' belief that they can self-monitor BP. The results have implications for theories of the mental representation of illness and for efforts to improve compliance with antihypertensive therapy. PMID- 10367612 TI - Ambulatory blood pressure responses and the circumplex model of mood: a 4-day study. AB - BACKGROUND: The relation between mood or emotions and concurrent ambulatory blood pressure responses holds both fundamental and clinical interest. METHODS: The primary sample consisted of 69 normotensive or borderline hypertensive but otherwise healthy adult males. The validation sample consisted of 85 healthy male undergraduate college students. Both samples underwent half-hourly 24-hour ambulatory blood pressure measurements on four separate workdays, 1 week apart. At each ambulatory measurement, subjects recorded their behavior, environment, and mood. The circular mood scale, a circular visual analogue scale based on the circumplex model of mood, was used to reflect the totality of a participant's affective state space. Longitudinal random effects regression models were applied in the data analysis. RESULTS: The results for both samples were quite similar. Sleep and posture had the greatest influence on ambulatory blood pressure and heart rate. The effects of the environmental setting, social setting, and consumption were modest but statistically significant. Independent of these covariates, mood exerted a significant effect on blood pressure and heart rate. Relative to the "mellow" default category, blood pressure increased both for "anxious/annoyed" and "elated/happy" and decreased during "disengaged/sleepy" mood. The range of mood-related blood pressure estimates was 6.0/3.7 mm Hg. CONCLUSIONS: The pattern of blood pressure responses suggests that they were related to the degree of engagement of a mood rather than the degree of unpleasantness. The hypothesis that posits that negative affect-related cardiovascular reactivity mediates the observed correlation between negative affect and disease risk should be reconsidered. PMID- 10367614 TI - Measuring counterdependency in patients with chronic pain. AB - OBJECTIVE: Some reports have characterized patients with chronic pain as counterdependent, that is, having emotional suppression, idealization of relationships, strong work ethic, a caregiver role-identity, and self-reliance. However, research has been hampered because formal measures of these traits have been lacking. In this article, we describe a five-item self-report questionnaire, the Counterdependency Scale (CDS), designed to elicit each of these traits on a Likert scale. METHODS: The CDS was administered to 150 consecutive patients evaluated in an outpatient psychiatry consultation program. RESULTS: CDS scores were normally distributed and had significant interitem correlations and test retest reliability (r = 0.68). As expected, subjects with chronic pain (N = 100) had higher mean CDS scores than those without chronic pain (t = 5.6, p = .000). CDS scores were independent of demographic variables and measures of anxiety, depression, alexithymia, and somatic amplification. CONCLUSIONS: These results suggest that counterdependency can be described by a distinct and measurable cluster of traits associated with chronic pain. PMID- 10367613 TI - Anxiety reduces baroreflex cardiac control in older adults with major depression. AB - OBJECTIVE: Although depression and anxiety predict risk of cardiac mortality, the contributions of depression and anxiety to vagal cardiac control have not been systematically evaluated. The goal of this study was to examine the relationship between state anxiety and vagal control of heart rate in older adults with major depressive disorder (MDD). Older adults (50-70 years old) were selected for this study because of the greater cardiac risk associated with low vagal cardiac control across this age range. METHODS: Fifty-six men and women with MDD were evaluated. MDD was diagnosed using the Diagnostic Interview Schedule, and severity of depression was measured using the Beck Depression Inventory and the Hamilton Rating Scale for depression. State anxiety was measured using the Spielberger State Anxiety Inventory. Power spectral analysis was used to measure two indices of vagal control: baroreflex control of heart rate (BRC(SPEC)) and respiratory sinus arrhythmia (RSA). RESULTS: State anxiety was negatively correlated with levels of BRC(SPEC) (r = -0.32, p < .05), whereas depression severity was not related to either RSA or BRC(SPEC). Furthermore, BRC(SPEC) was reduced by approximately 33% in MDD patients with state anxiety scores (ST-ANX) in the highest quartile (ST-ANX > 41, N = 13), compared with patients with ST-ANX scores in the lowest quartile (ST-ANX < 25, N = 14; p < .05). CONCLUSIONS: Anxiety, but not depression severity, is associated with reduced BRC(SPEC) in older men and women. Future studies are needed to determine whether comorbid anxiety contributes to the increased cardiovascular risk associated with MDD. PMID- 10367615 TI - Ethnic differences in thermal pain responses. AB - OBJECTIVE: Although numerous studies have reported ethnic differences in the prevalence and severity of clinical pain, little is known about how these differences affect the perception of experimental pain. The present experiment examined the effects of ethnicity (African American vs. white) on thermal pain responses in a healthy undergraduate population. METHODS: Thirty white subjects (16 women and 14 men) and 18 African Americans (10 women and 8 men) participated in the study. Thermal testing included evaluation of the following: warmth thresholds, thermal pain thresholds, thermal pain tolerances, and magnitude estimates of both the intensity and unpleasantness of thermal pain (at 46 degrees, 47 degrees, 48 degrees, and 49 degrees C). RESULTS: Although no group differences emerged for warmth thresholds, thermal pain thresholds, or pain intensity ratings, African Americans demonstrated lower thermal pain tolerances than whites. In addition, African Americans had smaller slopes and larger intercepts than whites for ratings of pain unpleasantness. Additional analyses suggested that these findings were a consequence of group differences in thermal pain unpleasantness ratings at the lowest temperatures assessed (46 degrees and 47 degrees C); at these temperatures, African Americans rated the stimuli as more unpleasant than whites. Finally, group differences in thermal pain tolerance and thermal pain unpleasantness ratings seemed to partially account for greater self reported daily pain symptoms among African Americans. CONCLUSIONS: Collectively, these findings seem to suggest ethnic differences in the perception of the affective-motivational dimension of thermal pain. PMID- 10367616 TI - Hypnosis in a case of long-standing idiopathic itch. AB - OBJECTIVE: This article presents the results of a brief hypnosis treatment of a woman with chronic, idiopathic vaginal and anal itch. METHODS: The patient was referred after 3 years of unsuccessful outcomes with standard topical and oral treatments prescribed by her family physician and three dermatologists. Treatment consisted of five sessions of self-hypnosis training in techniques of relaxation, deepening, and imagery, and home practice with an individualized instructional tape. RESULTS: After treatment, the patient reported substantial tissue healing, confirmed by her treating physician, that coincided with significant reductions in her scores of itch intensity, itch-related sleep disruption, and distress from pre- to posttreatment. These improvements continued at 4 months of follow-up, and the patient reported complete resolution of physical symptoms. CONCLUSIONS: The fact that these changes coincided with only minor improvements in general anxiety scores suggests that the resolution of the patient's itch condition was treatment specific rather than the result of methodological artifact, participant reporting bias, or a general sense of feeling better. These findings suggest that hypnosis is a cost-effective treatment for idiopathic itch conditions, especially those that are unresponsive to standard medical treatments. PMID- 10367617 TI - Development of a brief diagnostic screen for panic disorder in primary care. AB - OBJECTIVE: The purpose of this study was to determine the utility of a brief screening tool for panic disorder in the primary care setting. METHODS: A total of 1476 primary care outpatients in three primary care medical clinics on the West Coast of the United States were studied. Patients completed a brief self report measure, the five-item Autonomic Nervous System Questionnaire (ANS), while in the waiting room. The presence of DSM-IV panic disorder was subsequently determined in groups of "screen-positive" and "screen-negative" subjects using the Composite International Diagnostic Interview. A subset of patients (N = 511) also completed the 21-item Beck Anxiety Inventory. Indices of diagnostic utility were calculated using receiving operating characteristic analyses to guide the selection of optimal cutoff levels. RESULTS: The two-question version of the ANS had excellent sensitivity (range = 0.94-1.00 across the three clinic sites) and negative predictive value (0.94-1.00) but low specificity (0.25-0.59) and positive predictive value (range 0.18-0.40). The three- and five-question versions of the ANS had only modestly improved specificity, and this was achieved at the cost of reduced sensitivity and increased respondent burden to complete the questionnaire. The 21-item Beck Anxiety Inventory had maximal clinical utility at a cutoff level of > or =20, but sensitivity was lower than desirable for a screening instrument (0.67). CONCLUSIONS: The two-question version of the ANS shows promise as a screening instrument for panic disorder in the primary care setting. PMID- 10367618 TI - Endothelial function and hemodynamic responses during mental stress. AB - OBJECTIVE: The hemodynamic basis of blood pressure responses during psychological stress shows striking individual differences that share an interesting similarity with risk for cardiovascular disease. Factors accounting for these individual differences are poorly understood. The present study examined the relationship of vascular endothelial function to stress-induced hemodynamic responses. METHODS: Subjects were 40 healthy men and women, aged 25 to 44 years. Hemodynamic responses were assessed during exposure to a battery of four diverse laboratory stressors. Endothelium-dependent arterial dilation (EDAD) was measured by ultrasound imaging of the brachial artery in response to reactive hyperemia. RESULTS: High EDAD response was associated with lower resting systolic (p < .01) and diastolic blood pressure (p < .05). EDAD response was unrelated to blood pressure responses during stress. However, systemic vascular resistance responses during laboratory stress were significantly greater (p < .02) for individuals with low EDAD responses. CONCLUSIONS: Exaggerated systemic vascular resistance responses during stress may reflect endothelial dysfunction. This association may help explain the growing evidence of a relationship between stress hemodynamics and cardiovascular disease risk. The nature of this association is discussed in terms of a possible interplay between the sympathetic nervous system and the endothelium in regulation of vascular tone. PMID- 10367619 TI - Capacity to produce cytokines during weight restoration in patients with anorexia nervosa. AB - OBJECTIVE: Anorexic patients are surprisingly free of infectious complications despite their seriously undernourished state. To study this phenomenon, we longitudinally measured the capacity to produce cytokines in restricting-type anorexic patients. METHODS: Lymphoproliferative responses with phytohemagglutinin (PHA) and the capacity of whole blood to produce cytokines, such as interleukin-1 (IL-1), IL-6, tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN gamma), and granulocyte-colony stimulating factor (G-CSF), were longitudinally measured before and after weight gain, that is, at admission and at less than 60, 65, and 75% of standard body weight (SBW), in 17 patients with restricting-type anorexia nervosa and in 17 control subjects. RESULTS: Cytokine production of IL 1, IL-6, and TNF-alpha per monocyte in the anorexic patients recovered only with the start of refeeding, whereas IFN-gamma production per lymphocyte was similar to that in control subjects and did not change during weight restoration. Only G CSF production, even at 75% SBW, did not improve during weight restoration. Between the weight at admission and 65% SBW, the increase in the percentage of SBW and improvement of the total protein level were significantly correlated with improvement of the lymphocyte proliferative response with PHA. CONCLUSIONS: The capacity to produce most cytokines recovered with the start of weight gain; however, recovery was not correlated with weight gain. The results suggest that the capacity to produce cytokines in these anorexic patients was dependent on something other than the absolute value of body weight, such as the start of refeeding, the neuroendocrine system, or the autonomic nervous system. PMID- 10367620 TI - Higher abnormal leukocyte and lymphocyte counts 20 years after exposure to severe stress: research and clinical implications. AB - OBJECTIVES: Research suggests that individuals with posttraumatic stress disorder (PTSD) are more likely to develop medical conditions and other stress-related psychiatric disorders. Given these findings and others suggesting that PTSD victims may have altered neuroendocrine and immune systems, the hypothesis that Vietnam veterans with PTSD have abnormally high leukocyte and lymphocyte counts was tested. METHODS: The leukocyte and lymphocyte status of male Vietnam "theater" veterans with current partial posttraumatic stress (N = 286), anxiety (N = 274), and depression disorders (N = 192) were compared with those of Vietnam veterans without these disorders 20 years after military service (N = 2190), controlling for intelligence, race, age, income, education, type of enlistment, Vietnam volunteer status, region of birth, cigarette smoking, illicit drug use, body mass index, and alcohol consumption. Abnormal values were defined using standard laboratory reference ranges. Adjusted mean differences also were compared. RESULTS: Based on the results of two-tailed tests, PTSD-positive veterans are more likely to have adjusted leukocyte (OR = 1.83, p = .04) and T cell (OR = 1.82, p = .045) counts above the normal range and higher mean adjusted leukocyte (p = .042), lymphocyte (p = .01), T-cell (p = .008), and CD4 cell (p = .027) counts. Those with anxiety disorders have adjusted lymphocyte (OR = 1.68, p = .048) and T-cell (OR = 2.06, p = .011) counts above range. They also have test results indicating reactive delayed cutaneous hypersensitivity (OR = 1.77, p = .006), which suggests the presence of highly sensitized T-cell lymphocytes. Finally, depressed veterans are less likely to have B-cell counts above the reference range (OR = 0.55, p = .006). Results of one-tailed tests further suggest that PTSD-positive men also have abnormally high CD4 and CD8 T-cell lymphocyte counts as well (p < .05). CONCLUSIONS: Our findings suggest that chronic, primarily combat-related PTSD is associated with clinically elevated leukocyte and total T-cell counts. Those with current anxiety also have some of these abnormalities in addition to highly sensitized T-cell lymphocytes. Additional research is needed to specify the mechanisms involved here and to investigate the health risks associated with these findings. PMID- 10367621 TI - Ambulatory blood pressure, heart rate, and neuroendocrine responses in women nurses during work and off work days. AB - OBJECTIVE: This study examined women's cardiovascular and neuroendocrine responsiveness to work. METHODS: Ambulatory blood pressure (BP) and heart rate (HR) were recorded over 24-hour periods on 2 work and 2 off days during the luteal and follicular phases of the menstrual cycle in 138 registered nurses, aged 25 to 50 years. Urinary catecholamines and cortisol were measured for day and night periods. RESULTS: During waking hours systolic BP (SBP), HR, and epinephrine were higher on work than off days. Diastolic BP (DBP) and HR were highest at work. Nurses scoring high on job demands had elevations in daytime SBP, daytime HR only on work days, and nighttime epinephrine on work days. Compared with those with short work histories, nurses employed longer had consistently higher norepinephrine levels during days and nights, and higher nighttime DBP during off days. In unmarried nurses compared with married nurses, nighttime cortisol was lower during all 4 days and norepinephrine was lower during days off. All findings were independent of actigraph-recorded activity. CONCLUSIONS: Although the work environment leads to increased activity of the cardiovascular and sympathoadrenal medullary system in healthy women, the effects are modified by the woman's domestic role, by the length of her employment, and by the demands of her job. PMID- 10367623 TI - Studies of oncogenes and tumor-suppressor genes in human thyroid carcinomas, and their clinical implications. AB - INTRODUCTION AND DISCUSSION: Molecular genetic investigations relating to thyroid cancer have started to gain clinical importance, with discovery of the tumor specific oncogenes PTC 1-3 in papillary thyroid cancer and the syndrome-specific mutations of the RET protooncogene in MEN-2a, MEN-2b and familial MTC patients. Furthermore, the thyroid-specific sodium-iodine symporter, causing iodine accumulation in thyroid tissue, has been cloned and now offers studies to enhance iodine and radioiodine uptake in thyroid cancer, which could soon prove to be clinically applicable. PMID- 10367622 TI - Progression to AIDS: the effects of stress, depressive symptoms, and social support. AB - OBJECTIVE: We examined the effects of stress, depressive symptoms, and social support on the progression of HIV infection. METHODS: Eighty-two HIV-infected gay men without symptoms or AIDS at baseline were followed up every 6 months for up to 5.5 years. Men were recruited from rural and urban areas in North Carolina as part of the Coping in Health and Illness Project. Disease progression was defined using criteria for AIDS (CD4+ lymphocyte count of <200/microl and/or an AIDS indicator condition). RESULTS: We used Cox regression models with time-dependent covariates, adjusting for age, education, race, baseline CD4+ count, tobacco use, and number of antiretroviral medications. Faster progression to AIDS was associated with more cumulative stressful life events (p = .002), more cumulative depressive symptoms (p = .008), and less cumulative social support (p = .0002). When all three variables were analyzed together, stress and social support remained significant in the model. At 5.5 years, the probability of getting AIDS was about two to three times as high among those above the median on stress or below the median on social support compared with those below the median on stress or above the median on support, respectively. CONCLUSIONS: These data are among the first to demonstrate that more stress and less social support may accelerate the course of HIV disease progression. Additional study will be necessary to elucidate the mechanisms that underlie these relationships and to determine whether interventions that address stress and social support can alter the course of HIV infection. PMID- 10367624 TI - Compartment-mediated dissection for papillary thyroid cancer. AB - There is considerable controversy surrounding the appropriate treatment of papillary thyroid carcinoma (PTC), most of which centers around the extent of thyroidectomy. Despite the advocation of less than total thyroidectomy by many surgeons, there is a renewed interest by others, mainly in Europe and Japan, in the performance of routine total thyroidectomy and extensive lymph-node dissection for PTC. This has been shown to be an effective strategy for medullary thyroid carcinoma, which is not responsive to thyroid suppression or radioactive iodine treatment. PTC, however, is well treated by these adjuvant modalities and, in general, has an excellent prognosis. The benefit of extensive operations for routine cases of PTC has not been proven, and this practice is not employed by most surgeons in the United States. Node dissection is reserved for those patients with palpable adenopathy. PMID- 10367625 TI - C-cell cancer--prevention and treatment. AB - INTRODUCTION: C-cell cancer of the thyroid or medullary thyroid carcinoma (MTC) exists in a sporadic and a hereditary form, the latter of which is part of the multiple endocrine neoplasia type-2 (MEN-2) syndromes. DISCUSSION: MTC metastasises early to local (lymph nodes) and distant sites (liver, lung, bone). Therefore, early detection is mandatory to enable a chance of cure. In sporadic MTC, the sensitive tumour marker calcitonin enables detection of the disease at an early stage. In hereditary MTC, more than 95% of the patients have germline RET mutations. Thus, MEN-2 has become the paradigm for the practice of molecular medicine, and gene carriers can be identified before MTC even occurs. Surgery is the only chance of cure and recently developed surgical techniques provide the therapeutic prerequisite to achieve calcitonin normalisation in both sporadic and hereditary MTC. PMID- 10367626 TI - Risk factors associated with intraabdominal infections: a prospective multicenter study. Peritonitis Study Group. AB - INTRODUCTION AND METHODS: A prospective observational multicenter study with 18 hospitals was performed to assess preoperative risk, therapeutic management and outcome of patients with peritonitis. Data collection was carried out according to standardized and recommended definitions. Included in the study were 355 patients with macroscopically confirmed peritonitis. RESULTS: In the univariate analysis, the following factors influenced both the mortality and the incidence of postoperative complications: age, presence of certain concomitant disease, site of origin of peritonitis, type of admission and the ability of the surgeon to eliminate the source of infection. In addition, postoperative infective complications were related to the etiology of peritonitis and the exudate. In the multivariate analysis, APACHE II (P<0.001), successful operation (P<0.001), age (P<0.001), liver disease (P<0.03), malignant disease (P<0.04) and renal disease (P<0.05) turned out to be significant with respect to death. Escherichia coli was the predominant organism (51%), following by enterococci (30%) and bacteroides (25%). There was a significantly higher postoperative infection rate in patients with no adequate treatment of enterococci than patients with adequate treatment or no enterococci (P<0.05). CONCLUSION: The study demonstrated the important role of the physiological reserve of the patient and of the surgeon, which is not adequately reflected in existing scoring systems. Further investigations are needed to study the impact of enterococci on the outcome. PMID- 10367627 TI - Procalcitonin as a marker of severity in septic shock. AB - BACKGROUND/AIMS: Procalcitonin (PCT) was shown to be related to the severity of bacterial infection and is recommended as a new parameter of inflammation and infection. To evaluate the prognostic value in septic shock, PCT levels were repeatedly determined and compared with tumour necrosis factor-alpha (TNF-alpha)- and interleukin (IL)-6 bioactivity as well as with C-reactive protein (CRP) serum levels. PATIENTS: Twenty-four surgical patients with septic shock were included. Eight patients died within the study period of 14 days. METHODS: Serum levels of TNF-(WEHI 164) and IL-6 (B13-29 subclone 9) bioactivity, CRP and PCT were determined on days 1, 3, 5, 7, 10 and 14 following diagnosis of septic shock. RESULTS: Survivors and non-survivors were comparable in terms of age and severity of sepsis characterized by the APACHE II score and multiple-organ-failure score. Predominant causes of sepsis were peritonitis and necrotiszing pancreatitis. TNF levels increased in non-survivors with no significant difference to survivors. IL 6 bioactivity was increased on day 1 (P = 0.06) and remained elevated in non survivors, in whom it was significant on day 7 (P<0.05). CRP was constantly elevated with no difference between the groups. In nonsurvivors PCT remained increased, while the course of survivors was characterized by decreased values which were significantly lower (P<0.05) at every time point compared with those patients who died. A significant correlation could be found on day 1 (P<0.05) and at the end of the observation period (P<0.01) when comparing PCT levels with the multiple-organ-failure score. CONCLUSIONS: PCT seems to be a more reliable prognostic parameter in septic shock than IL-6, while TNF and CRP did not show any difference between survivors and non-survivors. These data indicate that PCT may represent a valuable parameter not only in the diagnosis of sepsis but also in the clinical course of the disease. PMID- 10367628 TI - Solid-pseudopapillary tumor of the pancreas--a rare and frequently misdiagnosed neoplasm. AB - INTRODUCTION: We describe a young woman with an unusual pancreatic tumor. FINDINGS AND DISCUSSION: Intraoperatively, a smoothly demarcated and encapsulated tumor was exposed. It was large (5 cmx4 cm) and of solid consistency, with a small stalk attached to the uncinate process. The tumor was partially surrounded by the pancreatic head. The macroscopic appearance suggested a benign tumor. Frozen sections revealed a benign pancreatic tumor, most likely of endocrine nature. Based on these findings, tumor enucleation was performed. The patient recovered rapidly from the intervention and was discharged from hospital after 2 weeks. One year after surgical treatment, the patient is without recurrence. The final diagnosis of the tumor was a solid pseudo-papillary tumor. PMID- 10367629 TI - Cystic neoplasms of the pancreas--cystadenomas and cystadenocarcinomas. AB - INTRODUCTION: Among the rare cystic pancreatic tumors, serous and mucinous cystadenoma and mucinous cystadenocarcinoma are most often diagnosed. CASE: We report on a total of 21 patients with cystic neoplasms who underwent surgery, 11 of whom had mucinous cystadenocarcinoma. Of the 10 remaining patients, serous and mucinous cystadenoma were diagnosed in two groups of five. A common feature of all cystic neoplasms is slow growth, leading to clinical symptoms at an advanced stage, with tumors frequently becoming enormous. RESULTS: In approximately half of the cases, diagnosis was possible by means of ultrasound, computed tomography and, in three instances, by preoperative percutaneous aspiration. Differential diagnosis of pseudocysts proved to be most difficult. CONCLUSION: Given the low operative risk, resection should always be performed in instances where findings cannot be clearly identified. Moreover, compared with ductal pancreatic carcinomas, the prognosis of a cystadenocarcinoma after early resection is extremely favorable, so that postponing resection might reduce the patient's prospects of being cured. PMID- 10367630 TI - Complications associated with different surgical approaches to differentiated thyroid carcinoma. AB - INTRODUCTION: With the good prognosis associated with differentiated carcinoma, the morbidity and mortality of different surgical approaches are of crucial importance. METHODS: At the Department of Surgery (Virchow Klinikum Berlin), 139 patients who underwent surgery for differentiated thyroid carcinoma between 1979 and 1994 were reviewed, focussing on postoperative complications. In 113 and 18 patients, respectively, primary and completion thyroidectomy was performed. In five patients, less than total thyroidectomy and in three patients only palliative surgery was carried out. We performed thyroidectomy without systematic lymphadenectomy (LAD) in 70 patients (51.1%). In 15 patients (10.8%), lymphadenectomy of the lateral compartment and, in 53 patients (38.1%), central LAD was performed. LAD did not significantly influence survival time in either follicular (n = 42) or papillary carcinoma (n = 97). RESULTS: No patient died because of postoperative complications. Permanent laryngeal nerve palsy occurred in no patients after thyroidectomy without LAD, in one patient after central LAD (1.9%) and in one patient after lateral LAD (6.7%). Transient laryngeal nerve palsy was seen in ten patients [six (8.6%) after thyroidectomy only, two (3.7%) after central LAD and two (13.3%) after lateral LAD] (P = 0.19). Hypocalcemia was distributed equally within the LAD groups: total transient hypocalcemia could be recorded in 54 patients (38.8%), but permanent hypocalcemia occurred only in one patient (0.7%). Postoperative recovery was delayed in patients when a more radical approach was used (P = 0.03). CONCLUSION: The magnitude of the benefit of LAD in therapy for differentiated thyroid carcinoma is still controversial. This more radical approach is not necessarily accompanied, however, by higher morbidity and mortality. PMID- 10367631 TI - Expression of wild-type ret, ret/PTC and ret/PTC variants in papillary thyroid carcinoma in Germany. AB - INTRODUCTION: Papillary thyroid carcinoma (PTC) is the most common thyroid malignancy. Although the clinical course is usually rather benign, a subset of tumors is more aggressive. The ret/PTC oncogene was found only in PTC, with varying frequencies of up to 30%. Recently, two new variants of ret/PTC could be identified in post-Chernobyl PTCs, which raised the possibility that the prevalence of ret/PTC in non-radiation-induced PTCs might be higher than previously described. Normal thyroid cells do not express wild-type ret, but there is evidence that ret activation from any cause, including wild-type ret, occurs in more than a half of papillary tumors. METHODS: We used reverse transcription polymerase chain reaction and sequencing to examine wild-type ret and all five forms of ret/PTC known today in 99 PTCs from Hannover, Dusseldorf, Halle and Regensburg. Our method could also detect other variants within the known breakpoint regions. The presence of the ret tyrosine-kinase domain was examined by immunohistochemistry. RESULTS: Seven PTC1-positive tumors and one PTC3-positive tumor (8%), but none with the new variants or other variants of PTC1, 2 or 3 could be detected. Of 43 tumors examined, 20 showed expression of wild-type ret mRNA and staining of ret protein located predominantly to the cell membrane. CONCLUSION: Variants of ret/PTC do not substantially contribute to non radiation-related ret/PTC-positive tumors, and the prevalence of ret/PTC in Germany is low in contrast to the high rate of wild-type ret expression. Thus, expression of wild-type ret should be examined for pathogenic, prognostic and possible therapeutic implications. PMID- 10367632 TI - No correlation between RET immunostaining and the codon 918 mutation in sporadic medullary thyroid carcinoma. AB - INTRODUCTION: Medullary thyroid carcinoma (MTC) occurs sporadically or as part of the inherited cancer syndrome, multiple endocrine neoplasia (MEN) type 2. The MEN2 gene has been identified as the RET proto-oncogene. Mutations in the RET proto-oncogene are associated with the pathogenesis of MTC. Approximately 23-40% of sporadic MTCs (sMTCs) have a somatic RET codon 918 mutation within the catalytic core of the tyrosine kinase, which is a mutation found in over 98% of all MEN 2B cases as a germline mutation. METHODS: In order to elucidate the role of this mutation, we examined 40 sMTCs for the codon 918 mutation. Simultaneously, we looked for overexpression of the RET protein by means of immunohistochemistry with a newly developed RET antibody. RESULTS: In 8 of 40 tumors (20%), we were able to find a RET codon 918 mutation. Nine of 40 tumors (22.5%) showed immunoreactivity with the RET antibody. CONCLUSION: The presence of the somatic RET codon 918 mutations did not correlate with the presence of positive RET immunostaining. PMID- 10367633 TI - Involvement of endothelin/nitric oxide balance in hepatic ischemia/reperfusion injury. AB - INTRODUCTION: Endothelin (ET) and nitric oxide (NO) act as opponents in the regulation of the hepatic microcirculation. During ischemia/reperfusion (I/R) ET levels are increased, whereas no rise in NO levels is observed. This imbalance may be responsible for microcirculatory disturbances. The aim of this study was to restore the delicate ET/NO balance to maintain the integrity of the hepatic microcirculation and to reduce I/R injury. METHODS: Ischemia was induced by crossclamping of the hepatoduodenal ligament for 30 min with portal decompression using a splenocaval shunt (56 Wistar rats, 200-250 g). Sham operation, ischemia and treatment groups with the endothelin receptor antagonist (ERA) bosentan (1 mg/kg body weight i.v.) and the NO donor L-arginine (400 mg/kg body weight i.v.) were performed. For assessment of the microcirculation, sinusoidal perfusion rate, diameters of sinusoids and postsinusoidal venules, leukocyte endothelium interactions and velocity of free-flowing leukocytes were investigated by means of in vivo microscopy 30-90 min after reperfusion. Local hepatic tissue PO2 was measured prior to ischemia, 30 min and 60 min after reperfusion and aspartate aminotransferase (AST)/alanine aminotransferase (ALT) levels were investigated 2 h and 6 h after reperfusion. RESULTS: After ischemia, sinusoidal and venular diameters were reduced to 76% and 85%, respectively, of sham operation group values (P<0.05), but were maintained at baseline in ERA (98/102%) and NO (102/105%) groups (P<0.05). Increased postischemic leukocyte sticking in sinusoids (144%) and venules (435%) was reduced by therapy to 110/253% (ERA) and 111/ 324% (NO), respectively (P<0.05). Perfusion rate was increased to 93% and 94% compared with 82% in the ischemia group (P<0.05). Concomitant with the improved microcirculation in therapy groups, local hepatic tissue pO2 was improved 30 min after reperfusion in the ERA (11.0 mmHg) and the L-arginine group (11.5 mmHg) relative to the ischemic group (6.9 mmHg) (P<0.05). In addition, postischemic AST/ALT increase was reduced by therapy. CONCLUSION: Our results indicate that maintenance of ET/NO balance by blocking ET receptors, as well as providing a NO donor, protects the liver microcirculation and reduces hepatic I/R injury. PMID- 10367634 TI - Is mesh fixation necessary in abdominal hernia repair? Results of an experimental study in the rat. AB - BACKGROUND: Abdominal hernia repair with implantation of synthetic meshes using the sublay technique has resulted in low recurrence rates and high patient satisfaction. AIM: The purpose of this experimental animal study was to investigate whether mesh fixation is necessary in abdominal hernia repair using a polypropylene mesh in the sublay technique. METHODS: Forty-five rats were divided into three groups after creating an abdominal wall defect (CG control group, no mesh implantation; NoFixG mesh implantation without fixation group; SG mesh with suture fixation group) with 15 animals in each group. End-points were clinical herniation pressure, hydroxyproline (HP) concentration, mesh shape and number of fibroblasts/collagen fibres of the anchor zone 7, 14 and 90 days after implantation. RESULTS: Herniation pressure, HP content and number of fibroblasts were similar between NoFixG and SG, although significantly higher in these groups than in the CG (P<0.05). Both mesh groups had significantly higher counts of fibroblasts and collagen fibres than the CG. Mesh shrinking occurred in both groups but was less in the SG. CONCLUSION: Mesh fixation was not mandatory in abdominal hernia repair using this animal model. PMID- 10367635 TI - Gastric tonometry accurately predicts mortality in experimental peritonitis in both laparoscopic and conventional surgery. AB - OBJECTIVES: Tonometry is widely used in the diagnosis of sepsis and splanchnic ischemia. This study was devised to analyze the predictive value of gastric tonometry for outcome of experimental viscus perforation-induced peritonitis. The impact of conventional and laparoscopic intervention on tonometric measurements was the main scope. METHODS: This randomized controlled intervention trial was performed in a University experimental laboratory, using 24 female Duroc pigs. Pigs were subjected to gastric perforation followed by a 12 h interval of peritonitis, and then to either laparoscopic or conventional surgical repair of the defect with peritoneal lavage. Gastric tonometry and cardiocirculatory monitoring were performed. RESULTS: Septic shock associated with peritonitis and subsequent lethal outcome was accurately predicted with gastric tonometry. Changes of gastric mucosal pH correlated significantly with decreases of MAP (r2=0.880; P<0.001) and SVR (r2=0.678; P<0.001), increase of QT (r2=0.486; P=0.013), and mortality (r=0.752; P<0.001). Mortality was significantly higher in laparoscopically treated animals compared to those subjected to the open procedure (78% vs 22%; P<0.045). CONCLUSIONS: Gastric tonometry accurately predicted mortality in experimental peritonitis. The decline of gastric mucosal pH in the laparoscopic group was more than double that of to conventionally treated animals. This finding not only reflected the increase of systemic CO2 due to higher absorption during CO2-pneumoperitoneum, but probably also indicated a more severe form of splanchnic ischemia during laparoscopic surgery. Even though tonometry can be used to accurately predict mortality and separate the high risk group, extreme caution should be applied under conditions associated with severe peritonitis. PMID- 10367636 TI - Fascioliasis observed during laparoscopic cholecystectomy. AB - Fascioliasis is an uncommon zoonotic disease caused by Fasciola hepatica, a liver fluke, for which humans act as an accidental host, infected by the ingestion of water or raw aquatic vegetables contaminated with the metacercaria. We report the case of a patient who presented to our clinic with right upper abdominal pain and nausea. Physical examination and abdominal ultrasonography revealed cholelithiasis. Peripheral blood eosinophilia was the only positive sign observed during routine laboratory tests. We therefore decided to perform laparoscopic cholecystectomy. During laparoscopy peritoneal implants approximately 0.5-1 cm diameter were detected which gave an impression of peritoneal carcinomatosa. Laparoscopic cholecystectomy was performed, and biopsies were taken from the peritoneal implants which were examined histopathologically, and fascioliasis was determined. PMID- 10367637 TI - Temporary closure of full-thickness abdominal-wall defects with mesh grafts. AB - INTRODUCTION: A simple technique is presented here for temporarily covering massive, full-thickness, abdominal-wall defects, when they cannot be closed directly. METHODS: The exposed viscera can be covered with a meshed split thickness skin graft to close the wound and seal off the abdominal cavity from the outside. Once the patient's general condition improves, the epidermal layer of the mesh graft should be removed by dermabrasion to minimize the risk of epidermal cysts, and the defect should be closed either by primary closure or by a local or free flap, depending on the defect size. PMID- 10367638 TI - Amphibian pain and analgesia. AB - Analgesics are often not provided to amphibians because the presence and severity of pain may not be recognized in these animals. In addition, there is little information on the mechanism of action of analgesic agents in amphibians. However, amphibians possess appropriate neurologic components for transmitting pain from peripheral receptors to the central nervous system and antinociceptive mechanisms to modulate pain. They are capable of displaying behavioral and physiologic modification of pain systems in response to analgesic pharmacologic agents. Therefore, pain perception in amphibians is likely analogous to that in mammals and invasive, potentially painful procedures should be accompanied by appropriate analgesia and anesthesia. Although specific doses have not been established in clinical trials, basic research into the mechanisms and regulation of endogenous opioid systems demonstrates the potential clinical benefit for the use of opioids in these animals. Other analgesics such as alpha2-agonists, ketamine, and tricaine methanesulfonate have also demonstrated analgesic potential. PMID- 10367640 TI - Health evaluation of free-ranging rockhopper penguins (Eudyptes chrysocomes) in Argentina. AB - As part of annual colony counts in Santa Cruz Province, Argentina, a health survey of rockhopper penguins (Eudyptes chrysocomes) was conducted in 1994. Forty five birds were examined during handling procedures, and blood and fecal samples were collected for laboratory analysis. All birds appeared to be in good condition. No ecto- or endoparasites were found. Hematology, plasma chemistry, and plasma mineral levels were measured and correlated with the results of bacterial and viral serology. Antibodies against Chlamydia sp., avian adenovirus, avian encephalomyelitis virus, infectious bronchitis virus, avian reovirus, and paramyxovirus-1, -2, and -3 were found. Mean plasma chemistry and mineral values differed between individuals testing positive and negative on serologic tests. There was no serologic evidence of exposure to avian influenza virus, duck viral enteritis, infectious bursal disease, infectious laryngotracheitis, Aspergillus sp., or Salmonella pullorum. Trace amounts of endrin were found in the plasma of one bird, but all other chlorinated pesticide and polychlorinated biphenyl levels were below detectable limits. PMID- 10367639 TI - Processing postmortem specimens with C18-carboxypropylbetaine and analysis by PCR to develop an antemortem test for Mycobacterium avium infections in ducks. AB - Mycobacterium avium is the causative agent of the avian mycobacteriosis commonly known as avian tuberculosis (ATB). This infection causes disseminated disease, is difficult to diagnose, and is of serious concern because it causes significant mortality in birds. A new method was developed for processing specimens for an antemortem screening test for ATB. This novel method uses the zwitterionic detergent C18-carboxypropylbetaine (CB-18). Blood, bone marrow, bursa, and fecal specimens from 28 ducks and swabs of 20 lesions were processed with CB-18 for analysis by smear, culture, and polymerase chain reaction (PCR). Postmortem examination confirmed nine of these birds as either positive or highly suspect for disseminated disease. The sensitivities of smear, culture, and PCR, relative to postmortem analysis and independent of specimen type, were 44.4%, 88.9%, and 100%, respectively, and the specificities were 84.2%, 57.9%, and 15.8%, respectively. Reductions in specificity were due primarily to results among fecal specimens. However, these results were clustered among a subset of birds, suggesting that these tests actually identified birds in early stages of the disease. Restriction fragment length polymorphism mapping identified one strain of M. avium (serotype 1) that was isolated from lesions, bursa, bone marrow, blood, and feces of all but three of the culture-positive birds. In birds with confirmed disease, blood had the lowest sensitivity and the highest specificity by all diagnostic methods. Swabs of lesions provided the highest sensitivity by smear and culture (33.3% and 77.8%, respectively), whereas fecal specimens had the highest sensitivity by PCR (77.8%). The results of this study indicate that processing fecal specimens with CB-18, followed by PCR analysis, may provide a valuable first step for monitoring the presence of ATB in birds. PMID- 10367641 TI - Pharmacokinetics of ceftazidime in loggerhead sea turtles (Caretta caretta) after single intravenous and intramuscular injections. AB - The pharmacokinetics of ceftazidime in yearling loggerhead sea turtles (Caretta caretta) following single i.m and i.v. injections were studied. Eight juvenile 1.25+/-0.18 kg turtles were divided into two groups. Four animals received 20 mg/kg of ceftazidime i.v. and four received the same dose i.m. Plasma ceftazidime concentrations were analyzed by reverse-phase high-performance liquid chromatography. The i.v. and i.m. administration half-lives were 20.59+/-3.24 hr and 19.08+/-0.77 hr, respectively. The volume of distribution was 0.42+/-0.07 L/kg, and the systemic clearance was 0.217+/-0.005 ml/min/kg. Ceftazidime was detected in all blood samples and its concentration exceeded the minimum inhibitory concentration for Pseudomonas for 60 hr after i.m. and i.v. injections. PMID- 10367642 TI - Clarithromycin pharmacokinetics in the desert tortoise (Gopherus agassizii). AB - Clarithromycin is a new, safe orally administered macrolide antibiotic active against Mycoplasma sp. in humans. Single-dose and multidose pharmacokinetic parameters were determined for clarithromycin in wild-caught desert tortoises (Gopherus agassizii) seropositive for M. agassizii. Clarithromycin blood levels were measured in three tortoises for up to 72 hr after a single oral dose of 7.5 mg/kg. In a second group of six tortoises, levels were measured after a dose of 15 mg/kg. Noncompartmental iterative two-stage Bayesian and nonparametric expectation maximization pharmacokinetic parameters were determined for each animal assuming first order rate constants. At 15 mg/kg, the maximum concentration was 1.37 microg/ml, the time to maximum concentration was 8.0 hr, and a plasma half-life of 11.69 hr was derived from the latter method. The absorption constant was 0.08/hr, the absorption half-life was 8.47 hr, and the weight-normalized volume of distribution was 5.30 L/kg. Predictions derived by the latter method suggested a dosage of 15 mg/kg p.o. every 24 hr to achieve maximal blood levels of > or =1 microg/ml for multiple dosing. However, results from a preliminary multidose study with three tortoises indicate that the drug is accumulated; therefore, the predicted dose may be closer to 15 mg/kg p.o. every 2 3 days to maintain blood levels of 2-7.5 microg/ml. (For n = 3, 2-point linear regression median estimates for the apparent elimination rate constant (K) and half-life are 0.0227/hr and 30.52 hr, respectively.) This multidose accumulation reflects a slower apparent elimination than that predicted in the eight single dose tortoises (i.e., K = 0.0593/hr, t1/2 = 11.69 hr). This study highlights a potential pitfall of depending solely on single-dose studies and the potential value of oral administration in reptiles. PMID- 10367643 TI - A three-year study of viable airborne fungi in the North Carolina Zoological Park R.J.R. Nabisco Rocky Coast Alcid Exhibit. AB - Aspergillosis is a common cause of mortality in captive birds, particularly in recently imported birds or captive chicks and their parents. Use of the Andersen N-6 single-stage viable air sampler in the North Carolina Zoological Park (NCZP) R.J.R. Nabisco Rocky Coast Alcid Exhibit before and after the introduction of birds allowed a unique study of the mycological content of the air in a developing self-contained ecosystem. The Alcid Exhibit had a median count of 17 colony-forming-units (CFU)/m3 of air in comparison to 200-500 CFU/m3 and 1,000 3,500 CFU/m3 reported in human dwellings and the NCZP R.J. Reynolds Forest Aviary, respectively. Cladosporium and Penicillium represented 21.3% and Aspergillus 1.08% of the fungi collected. During the study, no respiratory mycoses were reported in any of the alcids. Continuous high-efficiency particulate air filtration, maintenance of low exhibit air temperatures, and an environment with little residual organic material capable of supporting fungal growth were important factors contributing to low colony counts. All colony counts >100 CFU/m3 in the exhibit were related to the apparent introduction of fungi from outside the facility. A reduction in the number of fungi transported from an external source into enclosed cool-temperature aviaries may be sufficient to avoid outbreaks of aspergillosis. PMID- 10367644 TI - Evaluation of urinary and serum metabolites in Asian small-clawed otters (Aonyx cinerea) with calcium oxalate urolithiasis. AB - Baseline renal function data was collected during 24-hr periods of feeding and fasting from three male and three female adult Asian small-clawed otters (Aonyx cinerea) with calcium oxalate urolithiasis. Urine was analyzed for calcium, phosphorus, and oxalate, and urinalyses were performed. There was no evidence of glucosuria, which has been previously reported in Asian small-clawed otters with urolithiasis. Urinary oxalate levels were quite high when compared with those of dogs and humans without uroliths, and the ratio of urinary oxalate to calcium was close to 1:1 during periods of food consumption. There was no significant difference in urinary oxalate excretion between the fed and fasting states. Urinary calcium excretion was five times greater during feeding than during fasting. Calcium levels were higher in the otters than those reported for dogs without uroliths but were similar to those for normal humans. Water consumption and urine production were significantly higher during periods of food consumption. Serum chemistry analyses and electrolyte levels were also determined. There was no evidence of hypercalcemia. Fractional clearance of calcium and phosphorus and endogenous creatinine clearance were significantly higher during food consumption than during fasting. Parathyroid hormone levels were similar to those reported for dogs and cats. Serum 25-hydroxy-vitamin D was slightly lower in the otters than in dogs. PMID- 10367645 TI - Sevoflurane anesthesia in desert tortoises (Gopherus agassizii). AB - The effects of sevoflurane on anesthesia induction, recovery, ventricular pressures, heart rate, ventricular pH, blood gas values, and electrolytes were evaluated in desert tortoises (Gopherus agassizii). Tortoises were orotracheally intubated while awake and ventilated manually with 3-7% sevoflurane in oxygen (1 L/min) to achieve desired expired sevoflurane concentrations. Data, consisting of induction time, recovery time, systolic, diastolic, and mean ventricular pressures, heart rate, ventricular pH, blood gas values, and electrolytes, were collected prior to anesthesia and sequentially at 2.50% and 3.75% expired sevoflurane as measured at the junction of the endotracheal tube and the breathing circuit. Blood pressure was measured and blood samples were collected through a 25-ga needle passed through a cardiac access port that was placed while the tortoises were in dorsal recumbency. Mean (+/-SE) induction time was 2.55+/ 0.55 min, recovery time was 27.58+/-7.55 min, and duration of anesthesia was 105+/-12 min. Mean (+/-SD) values for systolic, diastolic, and mean ventricular pressures in awake tortoises were 28+/-3 mm Hg, 22+/-2 mm Hg, and 24+/-2 mm Hg, respectively. Sevoflurane (2.5% expired) significantly decreased systolic (14+/-3 mm Hg), diastolic (12+/-1 mm Hg), and mean (13+/-1 mm Hg) ventricular pressures compared with those of awake tortoises. Ventricular pressures did not decrease further with increasing depth of anesthesia. Heart rate (32+/-4 beats/min) did not change significantly under sevoflurane anesthesia. Sevoflurane administration increased ventricular PO2 but did not change Na+, K+, or iCa++ concentrations. Sevoflurane appears to provide safe and effective anesthesia with rapid induction and recovery. PMID- 10367646 TI - Effects of in vitro hemolysis on serum biochemistry values of the bottlenose dolphin (Tursiops truncatus). AB - The effects of in vitro hemolysis on 23 biochemical analytes were assessed in sera from 14 clinically healthy Atlantic bottlenose dolphins (Tursiops truncatus). Each serum sample was divided into three portions for analysis: 1) nonhemolyzed control; 2) moderate hemolysis, simulated by adding hemolyzed serum to a final concentration of approximately 150 mg/dl Hb; and 3) severe hemolysis, simulated by adding hemolyzed serum to a final concentration of approximately 500 mg/dl Hb. Moderate hemolysis resulted in statistically significant increases in the mean values of iron, lactate dehydrogenase, potassium, and uric acid and a decrease in creatinine (P < 0.001). Severe hemolysis resulted in statistically significant changes in the mean values of the above analytes in addition to the following increases: alanine aminotransferase, calcium, and serum globulins (P < 0.001) and albumin and total protein (P < 0.01). Total bilirubin and gamma glutamyl transferase levels were lower in the severely hemolyzed sample (P < 0.001). Differences in mean values for alkaline phosphatase between nonhemolyzed and hemolyzed serum were not significant but did show a downward trend in the hemolyzed sera. The presence and severity of hemolysis must be considered in the interpretation of the serum chemistry values. PMID- 10367647 TI - Clinical problems of sloths (Bradypus sp. and Choloepus sp.) in captivity. AB - A 20-yr retrospective study of disease prevalence was carried out for 51 sloths (34 Bradypus sp. and 17 Choloepus sp.) at the Sao Paulo Zoo. A total of 81 clinical disorders were detected, including nutritional (45.7%), digestive (12.3%), and respiratory (12.3%) problems and injuries (6.1%). A definitive diagnosis was not possible in 8.6% of the cases. The incidence of disease varied according to seasonal climate (winter, 32.5%; spring, 24%; summer, 22.9%; autumn, 20.5%), time in captivity (96.4% of diseases occurred within the first 6 mo and 3.6% occurred thereafter), and type of enclosure (quarantine cage, 96.4%; exhibition enclosure, 3.6%). Both young animals (86.7%) and adults (3.2%) were affected. Parasites were identified by fecal examination in 45.4% of animals with clinical illness (Ascaris sp., 80%; Coccidia sp., 20%). Bacteria such as Salmonella enteritidis, Escherichia coli, and Citrobacter freundii were isolated from feces and/or organs. The first 6 mo in captivity are critical for these animals. Proper management and early identification of medical conditions in captivity have implications for sloth population in the wild. PMID- 10367648 TI - Weights, hematology, and serum chemistry of free-ranging brown boobies (Sula leucogaster) in Johnston Atoll, Central Pacific. AB - Hematologic and serum chemistry values are reported for 105 brown boobies (Sula leucogaster) from Johnston Atoll, Central Pacific. Hematocrit, estimated total plasma solids, total and differential white cell counts, serum glucose, calcium, phosphorus, uric acid, total protein, albumin, globulin, aspartate aminotransferase, and creatinine phosphokinase were analyzed. Hematologic and serum chemistry values varied with age and sex. Values were compared with those of red-footed boobies and other tropical and temperate marine pelecaniforms. PMID- 10367649 TI - Comparison of in vitro tests for evaluation of passive transfer of immunoglobulins in giraffe (Giraffa camelopardalis). AB - Serum samples from captive giraffe (Giraffa camelopardalis) were tested to assess passive transfer of immunoglobulins using in vitro methods developed for domestic ruminants. Estimated immunoglobulin levels were compared using five tests (protein electrophoresis, total protein refractometry, zinc sulfate turbidity, glutaraldehyde coagulation, and sodium sulfite turbidity). A linear relationship was observed among total protein, gamma globulin (electrophoretic measurement), and immunoglobulin level based on spectrophotometric measurement of zinc sulfate turbidity. Nonquantitative assays also demonstrated statistical correlation with the quantitative methods. Using criteria similar to those established for domestic species, cutoff values for failure of passive transfer (FPT) were established for these tests in neonatal giraffe: 1) total protein <6.0 g/dl; 2) gamma globulin < 0.5 g/dl; 3) estimated immunoglobulin level < 1,000 mg/dl (zinc sulfate turbidity); 4) glutaraldehyde coagulation test negative; or 5) no visually detectable turbidity in 16% sodium sulfite or Bova-S negative. Retrospective examination of the medical histories showed a strong statistical association between animals designated as having FPT and those that were removed from their dams based on clinical assessment to be hand-reared. Application of these tests in the field should allow earlier detection and intervention for FPT in neonatal giraffe. PMID- 10367650 TI - Nutritional secondary hyperparathyroidism in free-living fledgling American crows (Corvus brachyrhynchos brachyrhynchos). AB - From June 1994 to June 1996, 18 fledgling American crows (Corvus brachyrhynchos brachyrhynchos) from multiple locations on Long Island, New York, were presented with signs of metabolic bone disease characterized by folding fractures of the proximal tibiotarsus. Plasma alkaline phosphatase levels were elevated, and the calcium/phosphorus ratio and 25-hydroxycholecalciferol (25-(OH)D3) levels were decreased. The histopathologic diagnosis was parathyroid hyperplasia and generalized osteodystrophia fibrosa. A diet low in bioavailable calcium and/or vitamin D3 was the probable cause. Also, exposure to xenobiotics could have contributed to the depression of 25-(OH)D3 levels. PMID- 10367651 TI - Brucella-induced abortions and infection in bottlenose dolphins (Tursiops truncatus). AB - Two bottlenose dolphins (Tursiops truncatus) aborted fetuses that died as a result of Brucella infection. Brucella placentitis occurred in both cases. Infected placenta and vaginal/uterine fluids may transmit Brucella species to other cetaceans. In a third case, an identical organism was cultured from lung necropsy tissue of an adult female T. truncatus. Microbiology, specific polymerase chain reaction, and pulsed-field gel electrophoresis results supported the designation of an additional genomic group(s), Brucella delphini, for isolates adapted to T. truncatus. Current serologic diagnostic tests reliable for known Brucella species are unreliable in detecting dolphin brucellosis. Our findings, together with previous reports, suggest that dolphin brucellosis is a naturally occurring disease that can adversely impact reproduction in cetaceans. The zoonotic significance of cetacean brucellosis is unknown, although the disease has not been reported in people who have frequent contact with dolphins. Further studies on the zoonotic aspects, distribution, prevalence, virulence, and impact of this disease in cetaceans and other marine mammal species are needed. PMID- 10367652 TI - Fatal mycotic dermatitis in captive brown tree snakes (Boiga irregularis). AB - Cutaneous fungal infections occurred in four captive brown tree snakes (Boiga irregularis). The ventral scales were most commonly affected, and lesions began as areas of erythema and edema with vesicle formation, followed by development of caseous brown plaques. Lesions usually started where ventral scales overlapped and spread rapidly. All snakes died within 14 days after clinical signs were first noted. The deaths of three of the snakes were directly attributable to the cutaneous disease; the other snake died from renal failure and visceral gout, most likely induced by gentamicin therapy. Histologically, lesions consisted of epidermal hyperplasia and hyperkeratosis, with foci of epidermal necrosis, intraepidermal vesicle formation, and subacute inflammation of the underlying dermis. These lesions were associated with bacteria and numerous septate, branched fungal hyphae within the epidermis and overlying serocelluar crusts. Hyphae that penetrated through the superficial surface of the epidermis often formed terminal arthroconidia. The same species of fungus was isolated in pure culture from the skin of three snakes, but fungal cultures were not performed on samples from the fourth snake. The fungus has been identified as the Chrysosporium anamorph of Nannizziopsis vriesii based on its formation of solitary dermatophytelike aleurioconidia and alternate and fission arthroconidia. The source of the fungus in this outbreak was not determined; however, the warm, moist conditions under which the snakes were housed likely predisposed them to opportunistic cutaneous fungal infections. PMID- 10367653 TI - Cecal inversion and subsequent colocolic intussusception in a red wolf (Canis rufus gregoryi). AB - A 2-yr-old female red wolf (Canis rufus gregoryi) presented with weight loss and diarrhea. Abnormal clinical pathology included low serum calcium, sodium, chloride, globulin, and albumin levels. Differential diagnosis included infectious enteritis, intestinal parasitism, inflammatory bowel disease, hepatic or renal disease, and malnutrition. The wolf was treated empirically, but did not improve. A second examination revealed persistent poor musculature and stool quality. Abdominal palpation revealed a firm mass; contrast radiography confirmed an intussusception. Exploratory laparotomy revealed a colocolic intussusception involving the cecum. Following reduction of the colocolic intussusception, cecal inversion (cecocolic intussusception) was identified. Because the cecal inversion could not be reduced, typhlectomy was performed through a colotomy incision. Bacterial culture of peritoneal fluid yielded two strains of Escherichia coli. Postoperatively, the wolf was placed on antibiotics and a soft diet. The diet was gradually returned to its normal formulation and the wolf progressively gained weight. Physical examination 7.5 mo following initial presentation revealed normal body weight and condition. To our knowledge, this is the first recorded incidence of cecal inversion with concurrent colocolic intussusception. PMID- 10367654 TI - Cranial edema associated with a protein-losing nephropathy in a golden-mantled flying fox (Pteropus pumilus). AB - An adult golden-mantled flying fox (Pteropus pumilus) was diagnosed with nephrotic syndrome on the basis of the findings of proteinuria, hypoalbuminemia, hypercholesterolemia, and cranial edema. Membranoproliferative glomerulitis and interstitial nephritis were confirmed antemortem by renal biopsy. The bat had received seven injections of oxytocin in the period immediately prior to presentation. The possible role of oxytocin in the development of the nephropathy is discussed. Supportive care and treatment with a single plasma transfusion, furosemide, and prednisone led to a gradual but complete resolution of the nephrotic syndrome in this animal. PMID- 10367655 TI - Rickets in two hand-reared polar bear (Ursus maritimus) cubs. AB - Two polar bear (Ursus maritimus) cubs born at the Denver Zoological Gardens were abandoned by a primiparous mother. Lethargic and extremely chilled, the cubs were transported to the zoo hospital for intensive care and hand rearing. Both cubs developed rickets. Dietary changes were instituted, and both cubs completely recovered with the exception of a bowed right femur (genu varum) in the female cub. PMID- 10367656 TI - Hypospadias in a polar bear (Ursus maritimus). AB - Corrective surgery on a 1.5-yr-old male polar bear (Ursus maritimus) with hypospadias included amputation of the vestigial penis, bilateral orchiectomy with scrotal ablation, and distal perineal urethrostomy. Hypospadias in other species is a congenital deformity that may be caused by extra- and intrauterine factors resulting in a disruption of the testosterone balance during urethral development, but the causative mechanism in this bear is unknown. The urethrostomy site was functional without complications 8 mo after surgery. PMID- 10367657 TI - Double aortic arch and persistent left vena cava in a white lion cub (Panthera leo). AB - A 4-mo-old female white lion (Panthera leo) cub was presented with a 2-wk history of persistent postprandial regurgitation, mild dyspnea, and poor weight gain. The cub was weak and thin but otherwise alert. Survey and contrast radiography revealed a large dilated esophagus cranial to the heart base, with an esophageal filling defect present at the level of the fourth thoracic vertebra. A vascular ring anomaly was tentatively diagnosed. Exploratory thoracotomy revealed a double aortic arch and a persistent left vena cava. The left aortic arch was ligated and divided, and recovery was uneventful. A single episode of regurgitation occurred within the first postoperative month, and the cub gained 5.5 kg in weight during the same time period. Neither double aortic arch nor persistent left vena cava has been reported in a nondomestic felid. PMID- 10367658 TI - Partial tracheal obstruction due to chondromas in ball pythons (Python regius). AB - Over a 9-mo period, three adult ball pythons (Python regius) (one male, two females) were evaluated for severe dyspnea. Partial obstructions of the tracheal lumen were identified radiographically and/or visualized with a 3.0-mm rigid laparoscope inserted into the tracheal lumen in all three snakes. Administration of systemic antibiotics and nebulization resulted in partial improvement of the dyspnea. In two snakes, the tracheal lesions were removed with a rigid laparoscope and a flexible biopsy instrument inserted into the tracheal lumen. The other snake died and was necropsied. Histologically, the lesions from two snakes were determined to be benign chondromas. The chondromas were composed of a variably disorganized chondroid matrix populated by quiescent, normal-appearing chondrocytes within lacunae, although the chondrocytes were increased in density compared with normal hyaline cartilage and contained rare mitotic figures. The tracheal masses in one snake grew by expansion, not invasion, and were focally continuous with a mineralized cartilage tracheal ring, suggesting a benign nature. This is the second report of tracheal chondroma in ball pythons. Tracheal chondromas are exceedingly rare in humans and domesticated animals, suggesting a possible predisposition of ball pythons for this neoplasm. PMID- 10367660 TI - Herpesvirus-associated papillomas in koi carp (Cyprinus carpio). AB - From January through November 1994, 32% (7/22) of koi carp (Cyprinus carpio) maintained in indoor aquariums developed proliferative cutaneous lesions that consisted of single to multiple 2-10-mm whitish to pink fleshy masses usually associated with fin rays. Although scaleless koi were more commonly affected (3/6) than were normally scaled koi (4/16), the difference in incidence rates was not significant (chi2 text, P > 0.05). Lesions typically resolved spontaneously in 1-3 wk, occasionally persisted for >3 mo, and recurred in several fish after 2 5 mo. Fish were otherwise asymptomatic. Wet mount preparations from lesions were densely cellular and consisted of hyperplastic epidermal cells of normal morphology without parasites or inflammatory cells. Histologically, biopsies were consistent with papillomas and were characterized by a marked benign epidermal hyperplasia without inclusion bodies or inflammatory infiltrate. Transmission electron microscopic examination revealed intranuclear and intracytoplasmic herpesvirus virions. Virus isolation attempts were unsuccessful. PMID- 10367659 TI - Lymphosarcoma with leukemic blood profile in a Savannah monitor lizard (Varanus exanthematicus). AB - A 6-mo-old male Savannah monitor lizard (Varanus exanthematicus) was presented for lethargy and anorexia of 7 days duration. Physical examination revealed a slightly raised subcutaneous mass (1 cm diameter) in the left scapular area. Fine needle aspiration cytology of the mass revealed a population of immature, pleomorphic lymphoid cells consistent with lymphosarcoma. A hemogram indicated marked leukocytosis (465,000 cells/microl) characterized by extreme lymphocytosis and many circulating lymphoid blast cells. The lizard was euthanized at the owner's request. Necropsy revealed severe hepatomegaly and multiple raised, ulcerated mural masses in the gastrointestinal tract. There were many raised, poorly demarcated tan foci in all the parenchymal organs. Histopathologic examination confirmed infiltration of all parenchymal organs by neoplastic lymphoid cells. Transmission electron microscopic examination failed to identify viruses within the neoplastic cells. A literature review revealed few reports of squamate leukemia and lymphosarcoma and none in Savannah monitor lizards. PMID- 10367661 TI - Suspected intestinal torsion in a blacktip reef shark (Carcharhinus melanopterus). AB - A blacktip reef shark (Carcharhinus melanopterus) with a history of recurrent eversion or prolapse of the valvular intestine, arrived at the Steinhart Aquarium in June 1996 and was placed in a mixed species shark display. The eversion soon reappeared, and the animal became thin and anorexic. The everted portion of the intestine was bitten by another shark and became infected. Treatment included force-feeding, manual reduction of the eversion, hydrocortisone suppositories held in place with purse-string sutures, and injections of enrofloxacin. All treatments ultimately failed, and the animal died in March 1997. Necropsy revealed that the intestine had separated into two segments at the distal portion of the pyloric stomach (duodenum); both ends were scarred and sealed off. It is suggested that a torsion (twisting along the long axis) of the intestine was responsible for the condition. PMID- 10367662 TI - Case 1. Gastric outlet obstruction caused by foreign bodies. PMID- 10367663 TI - Case 2. Cholelithiasis of the liver. PMID- 10367664 TI - Pharmacology and safety assessment of humanized monoclonal antibodies for therapeutic use. AB - The humanization of monoclonal antibodies has generated a class of therapeutic products with improved safety, longer half-lives, and greatly diminished immunogenicity. These engineered proteins are highly species specific and in many cases only cross-react in humans. Where there is cross-reactivity in nonhuman primates or other species, it is not always clear that the pharmacologic effects reflect the potential actions in human volunteers or patients. As with other biologic products, the profile of humanized monoclonal antibodies dictates the preclinical strategy. The preclinical programs for the 2 humanized monoclonal antibodies described here, anti-HLA-DR (Hu1D10) and anti-CD3 (HuM291), demonstrate several unique aspects that affected their preclinical development strategy. Hu1D10 binds to a posttranslational form of HLA-DR and recognizes this antigen in some but not all human and nonhuman primates. The second antibody, HuM291, cross-reacts with CD3 only in the chimpanzee, which is not an optimal test species. In addition, a marketed anti-CD3 product exists (OKT3), and in the preclinical development of our antibody during testing of efficacy and safety, we needed to focus on adverse effects that might be similar to those of OKT3. In these studies, the safety, pharmacokinetics, immunogenicity, and pharmacology (B- and T-cell depletion and recovery) of the 2 antibodies were evaluated. The focus in this review is on the safety and pharmacology testing and the current status of each drug. PMID- 10367665 TI - Preclinical development strategies for novel gene therapeutic products. AB - With over 220 investigational new drug applications currently active, gene therapy represents one of the fastest growing areas in biotherapeutic research. Initially conceived for replacing defective genes in diseases such as cystic fibrosis or inborn errors of metabolism with genes encoding the normal, or wild type, gene product, gene therapy has expanded into other novel applications such as treatment of cancer or cardiovascular disease, where the risk:benefit profiles may be more acceptable in relation to the severity of the disease. Different types of vectors, including modified retroviruses, adenoviruses, adenovirus associated viruses, and herpesviruses and plasmid DNA, are used to transfer foreign genetic material into patients' cells or tissues. Developing a toxicology program to determine the safety of these agents, therefore, requires a modified approach that encompasses the pharmacology and toxicity of both the gene product itself and the vector system used for delivery in the context of the application for the clinical trial. In general, the issues involved in designing and developing appropriate preclinical testing to determine the safety of these products are similar to those encountered for other recombinant molecules, including protein biotherapeutics. Limitations to some of the typical toxicology studies conducted for a traditional drug development program may exist for these agents, and nontraditional approaches may be required to demonstrate their safety. Many factors may affect the safety and clinical activity of these agents, including the route, frequency, and duration of exposure and the type of vector employed. Other safety considerations include quantitation of the duration and degree of expression of the vector in target and other tissues, the effects of gene expression on organ pathology and/or histology, evaluation of trafficking of gene-transduced cells or vector after injection, and interactions of the host immune system with the transduced cell population. Because of the unique concerns regarding each of these therapies, the Center for Biologics Evaluation and Research encourages sponsors to obtain toxicity data whenever possible while evaluating the pharmacologic activity of the vector in a species or animal model relevant to their clinical indication. Sponsors are encouraged to discuss preclinical study design and results with the Center during product development to facilitate early identification of safety concerns prior to entry of these novel agents into the clinical setting and to ensure an uninterrupted course of development while addressing issues required for licensure. PMID- 10367666 TI - Synthetic oligonucleotides: the development of antisense therapeutics. AB - Antisense therapeutics using synthetic oligodeoxynucleotides (ODNs) are currently being evaluated in clinical trials for cancer, inflammation, and viral diseases. These macromolecules afford a unique opportunity to treat disease at the molecular level. The specificity of these compounds is derived from the genetic code and Watson-Crick base pairing, utilizing an antisense paradigm for the inhibition of translation and the regulation of protein expression. Currently, most antisense ODNs in development contain a phosphorothioate (P=S) backbone. Additional modifications primarily involve the 2' position on the ribose or modification of the nucleotide linkages of the backbone. To date, no toxicities in animal models appear related to inhibition of the pharmacologic target, rather toxicities induced by P=S ODNs appear similar and are independent of pharmacologic target. In general, toxicities correlate well with pharmacokinetic or tissue distribution parameters. In primates, the primary acute effects are associated with complement activation and the systemic effects associated with accumulation of high concentrations of P=S ODNs in the kidneys. In rodents, the primary effect is an immune stimulation characterized by splenomegaly, lymphoid hyperplasia, and mononuclear cell infiltrates in multiple tissues. At extraordinarily high doses (15-50 times the targeted clinical doses), hepatocellular and renal tubular degeneration are evident in rodents. Second generation antisense compounds, new routes of administration, and new formulations appear to broaden and improve the application of antisense technology. PMID- 10367667 TI - Efficacy and concentration-response of murine anti-VEGF monoclonal antibody in tumor-bearing mice and extrapolation to humans. AB - The development of a neovascular supply (angiogenesis) is a major aspect of tumorigenesis. Recent work has indicated that vascular endothelial growth factor (VEGF) is a major regulator of angiogenesis. In vitro and in vivo studies have demonstrated that an anti-VEGF antibody is capable of suppressing the growth of human tumor cell lines. The following study was conducted in tumor-bearing nude mice to evaluate the concentration-response relationship of murine anti-VEGF monoclonal antibody (muMAb VEGF) so that an efficacious plasma concentration of the recombinant humanized form (rhuMAb VEGF) in cancer patients could be estimated. (This study was included in our Investigational New Drug application to support the clinical dosing regimen and projected human safety factors for the toxicology program.) Additionally, the growth dynamics of the tumors were evaluated as a function of dose to explore whether a mechanismic interpretation of tumor growth inhibition by muMAb VEGF is possible. On day 1, A673 human rhabdomyosarcoma cells (2 x 10(6) cells/mouse) were injected subcutaneously in 188 beige nude mice (16-24 g). Treatment with muMAb VEGF (0.05-5.0 mg/kg; n = 24/group), phosphate-buffered saline (n = 10), or anti-gp120 isotype-matched control antibody (5.0 mg/kg; n = 10) began 24 hr later. Each animal received intraperitoneal injections of test material twice weekly for 4 wk. Immediately prior to each dose, 2 mice from each muMAb VEGF group were selected randomly, and plasma was collected for pharmacokinetic evaluation; at the end of the study, samples were collected from all animals for pharmacokinetic evaluation. Tumor dimensions were recorded weekly, and at the end of the study, tumor weight and dimensions were recorded. Satisfactory tumor suppression in nude mice was achieved at muMAb VEGF doses of > or =2.5 mg/kg, where the average trough muMAb VEGF plasma concentration was 30 microg/ml (concentrations in individual animals >10 microg/ml). Assuming the pharmacokinetics of rhuMAb VEGF in patients will resemble the pharmacokinetics of a similar humanized anticancer monoclonal antibodies, a clinical dosing regimen was designed to maintain the rhuMAb VEGF plasma concentration in this efficacious range. This study shows an approach that can be used to estimate a human dosing regimen from preclinical pharmacokinetic/pharmacodynamic data. Because we have just initiated clinical trials with rhuMAb VEGF we cannot judge clinical outcome in relation to these preclinical predictions; nonetheless, it is hoped that by sharing our approach and thought processes with other investigators we can assist the discovery and development of anticancer therapeutics. PMID- 10367669 TI - Safety assessment of biotechnology-derived pharmaceuticals: ICH and beyond. AB - Many scientific discussions, especially in the past 8 yr, have focused on definition of criteria for the optimal assessment of the preclinical toxicity of pharmaceuticals. With the current overlap of responsibility among centers within the Food and Drug Administration (FDA), uniformity of testing standards, when appropriate, would be desirable. These discussions have extended beyond the boundaries of the FDA and have culminated in the acceptance of formalized, internationally recognized guidances. The work of the International Committee on Harmonisation (ICH) and the initiatives developed by the FDA are important because they (a) represent a consensus scientific opinion, (b) promote consistency, (c) improve the quality of the studies performed, (d) assist the public sector in determining what may be generally acceptable to prepare product development plans, and (e) provide guidance for the sponsors in the design of preclinical toxicity studies. Disadvantages associated with such initiatives include (a) the establishment of a historical database that is difficult to relinquish, (b) the promotion of a check-the-box approach, i.e., a tendancy to perform only the minimum evaluation required by the guidelines, (c) the creation of a disincentive for industry to develop and validate new models, and (d) the creation of state-of-the-art guidances that may not allow for appropriate evaluation of novel therapies. The introduction of biotechnology-derived pharmaceuticals for clinical use has often required the application of unique approaches to assessing their safety in preclinical studies. There is much diversity among these products, which include the gene and cellular therapies, monoclonal antibodies, human-derived recombinant regulatory proteins, blood products, and vaccines. For many of the biological therapies, there will be unique product issues that may require specific modifications to protocol design and may raise additional safety concerns (e.g., immunogenicity). Guidances concerning the design of preclinical studies for such therapies are generally based on the clinical indication. Risk versus benefit decisions are made with an understanding of the nature of the patient population, the severity of disease, and the availability of alternative therapies. Key components of protocol design for preclinical studies addressing the risks of these agents include (a) a safe starting dose in humans, (b) identification of potential target organs, (c) identification of clinical parameters that should be monitored in humans, and (d) identification of at-risk populations. One of the distinct aspects of the safety evaluation of biotechnology-derived pharmaceuticals is the use of relevant and often nontraditional species and the use of animal models of disease in preclinical safety evaluation. Extensive contributions were made by the Center for Biologics Evaluation and Research to the ICH document on the safety of biotherapeutics, which is intended to provide worldwide guidance for a framework approach to the design and review of preclinical programs. Rational, scientifically sound study design and early identification of the potential safety concerns that may be anticipated in the clinical trial can result in preclinical data that facilitate use of these novel therapies for use in humans without duplication of effort or the unnecessary use of animals. PMID- 10367668 TI - Regulatory decision strategy for entry of a novel biological therapeutic with a clinically unmonitorable toxicity into clinical trials: pre-IND meetings and a case example. AB - The following material was derived from a synthesis of case histories taken from investigational new drug (IND) applications and drug sponsors' experiences, utilizing fictionalized data to avoid any resemblance to any proprietary information; any such resemblance is accidental. These examples are used as an instructional scenario to illustrate appropriate handling of a difficult toxicology issue. In this scenario, a drug caused a toxicity in animals that was detected only by histopathologic analysis; if it were to develop in patients, no conventional clinical methods could be identified to monitor for it. It is not unusual for a firm to cancel clinical development plans for a lead drug candidate that causes such a toxicity, especially if such a drug is intended for use as a chronic therapeutic in a population of patients with a chronic disease. This case synthesis was inspired by a Food and Drug Administration (FDA) agreement to allow such a product to proceed into clinical trials after substantive pre-IND discussions and agreement on well-considered toxicology program designs. The scientists most closely involved in the strategy development included the sponsor's toxicologist, veterinary toxicologic pathologist, and pharmacokineticist, as well as the FDA's reviewing pharmacologist. The basis of this decision was thorough toxicity characterization (1-month studies in 2 species); correlating toxicities with a particular cumulative area under the curve (AUC) in both species; identification of the most sensitive species (the species that showed the lower AUC correlating with toxicity); allometric assessment of clearance of the drug in 3 nonhuman species; construction of a model of human kinetics (based on extrapolation from animal kinetics); and finally, estimation of clinical safety factors (ratios of the human estimated cumulative AUC at the proposed clinical doses, over the animal cumulative AUC that correlated with the no adverse effect levels). Industry and FDA scientists negotiated a joint assessment of risk and benefit in patients, resulting in the FDA permitting such a compound to enter into clinical trials for a serious autoimmune disease. Such constructive, early communication starts with the pre IND meeting, and the conduct and planning for this meeting can be very important in establishing smooth scientific and regulatory groundwork for the future of a drug under IND investigation. PMID- 10367670 TI - Flow cytometry in the preclinical development of biopharmaceuticals. AB - Novel biomarkers are often required in the preclinical development of biopharmaceuticals in order to characterize pharmacologic and toxicologic effects and to establish pharmacodynamic and pharmacokinetic relationships. Flow cytometry is uniquely suited for measurement of these biomarkers. Large numbers of single cells in a heterogeneous population can be rapidly identified and characterized with high accuracy and reproducibility. Cells are not damaged by the detection system and can be subsequently sorted for further morphologic or functional analysis. The availability of clinical instruments and a wide range of fluorescent probes have made this technology applicable for use in toxicologic clinical pathology. Flow cytometry has played an integral role in the development of a monoclonal antibody to human CD4 (keliximab, IDEC-CE9.1, SB 210396). Lymphocyte subset analysis and assays for expression, coating, and modulation of human CD4 were used for sequential assessment of the pharmacologic activity of keliximab in transgenic mice expressing human CD4. PMID- 10367671 TI - Lesions and identification of crystalline precipitates of glycoprotein IIb-IIIa antagonists in the rat kidney. AB - Two glycoprotein IIb-IIIa antagonists (xemilofiban, SC-54684A, and orbofiban, SC 57099B), which are platelet aggregation inhibitors, caused crystalline precipitates in the kidney tubules of rats at high dosages. Dogs were not affected. Depending on the degree of the precipitation, which was dosage dependent, and the location, which differed somewhat between the two compounds, the lesions varied from acute obstruction with tubule cell necrosis, nephron dilation, and sudden death with no inflammation to severe chronic pyogranulomatous inflammation. In order to understand the relevance of the lesions, it was important to identify the precipitates. This was technically challenging because the crystals were water soluble (dissolving in routine fixing and staining techniques) and were present in insufficient quantity to physically isolate. Techniques were devised to evaluate the crystals in situ in unstained frozen sections prepared without directly embedding the tissues in supporting medium, which interfered with the analyses. The crystals were analyzed in situ by infrared and Raman spectroscopy and time-of-flight secondary ion mass spectroscopy (TOF-SIMS). Uroliths found in the renal pelvis of one animal were analyzed by liquid chromatography/mass spectrometry. The resulting spectra showed that the crystals were the de-esterified acids of the parent compounds. This knowledge allowed us to predict that the crystalline precipitates would not be a hazard to humans because of the large multiples of the human dosage at which they occurred and because of differences in renal physiology between rats, dogs, and humans. PMID- 10367672 TI - Use of in vivo confocal microscopy to understand the pathology of accidental ocular irritation. AB - In vivo confocal microscopy (CM) provides a unique ability to section optically through living, intact tissues and organs to characterize qualitatively and quantitatively pathological changes in 4 dimensions (x, y, and z, and time). It involves the capture of real-time images without the need for excision, fixation and processing. In vivo CM principally has been used for evaluation of eyes in patients and laboratory animals but has potential application to studies of other tissues/organs. In vivo CM is being used in human ophthalmology clinics. It has been used as a research tool for quantitative, in situ measurement of corneal wound contraction, fibroblast migration, corneal endothelial cell migration, corneal epithelial cell size and desquamation following contact lens wear and surgery, and the assessment of corneal surface toxicity following application of commonly used ophthalmic preservatives. In vivo CM allows us to (a) characterize changes to a light microscopic (i.e., cellular) level; (b) quantify changes objectively: (c) conduct studies of injury and repair in the same animal and directly correlate microscopic changes to clinical observations over time as this technique is used in the living animal; and (d) conduct comparative studies in humans. Here we present a brief overview of in vivo CM and how we are using it to provide noninvasive, in situ qualitative and quantitative histopathologic characterization of accidental ocular irritation. Our intent is to provide an awareness of this relatively new methodology and one practical application of its use in research. The goal of our work is to provide objective, quantitative data for use in developing and validating mechanistically based in vitro replacement tests. PMID- 10367673 TI - Molecular pathology in the preclinical development of biopharmaceuticals. AB - Advances in cell and molecular biology have engendered a wide range of techniques that can be used to study the molecular events that underlie the cause of disease, thus producing a new field of study called "molecular pathology." These techniques can be either slide-based or non-slide-based (solution-based). The slide-based techniques include immunohistochemistry, in situ hybridization, and in situ polymerase chain reaction; pathologists play a unique role in the administration of these techniques because of their ability to interpret the end product (i.e., the slide). In this manuscript, we briefly discussed the use and impact of these slide-based techniques within all phases of drug development in the pharmaceutical industry. PMID- 10367675 TI - The toxicology of interleukin-12: a review. AB - Recombinant murine interleukin (IL)-12 (rmIL-12) exhibits antitumor, antiviral, and antimicrobial activities and can modify allergic inflammatory reactions in animal models. Recombinant human IL-12 (rhIL-12) is currently in clinical trials for treatment of cancer, asthma, and viral hepatitis. Principally a phagocyte derived cytokine, IL-12 targets natural killer cells and T lymphocytes, stimulating their activity and the secretion of interferon (IFN)-gamma. An understanding of the toxicology of IL-12, due in part to effects mediated by IFN gamma, has emerged from preclinical safety and mechanistic studies and initial clinical trials. Target organs common to several animal species and humans include the lymphohematopoietic system, intestines, liver, and lung. PMID- 10367674 TI - Development of a recombinant interleukin-4-Pseudomonas exotoxin for therapy of glioblastoma. AB - About 12,000 Americans are diagnosed with malignant astrocytoma each year. Despite surgery, radiotherapy, and chemotherapy, the prognosis of these patients remains poor. Targeted toxins based on the identification of novel antigens or receptors provide a promising new approach to treating cancer. We have identified one such cell surface protein in the form of interleukin (IL)-4 receptors (IL-4R) on human malignant astrocytoma. Normal brain tissues from frontal cortex and temporal lobe cortex do not express IL-4R. To target IL-4R, we generated a chimeric fusion protein composed of IL-4 and Pseudomonas exotoxin (IL4-PE). This toxin is highly cytotoxic to IL-4R-bearing human brain cancer cells. Preclinical toxicologic experiments were performed in mice, rats, and guinea pigs to determine an maximum tolerated dose. Intrathecal administration in cynomolgus monkeys produced high cerebrospinal fluid levels without any central nervous system or other abnormalities. When IL4-PE was injected into the right frontal cortex of rats, localized necrosis was observed at 1,000 but not < or =100 microg/ml doses. Intravenous administration of this biologic to monkeys produced reversible grade 3 or grade 4 elevations of hepatic enzymes in a dose-dependent manner. These results indicate that localized administration can produce nontoxic levels of IL4-PE that may have significant activity against astrocytoma. In vivo experiments with nude mice have demonstrated that IL4-PE has significant antitumor activity against human glioblastoma tumor model. Intratumor administration of IL4-PE has been initiated for the treatment of malignant astrocytoma in a phase I clinical trial. PMID- 10367676 TI - Preclinical and early clinical development of keratinocyte growth factor, an epithelial-specific tissue growth factor. AB - Keratinocyte growth factor (KGF) is a 28-kDa heparin-binding member of the fibroblast growth factor (FGF) family (alternative designation = FGF-7) that specifically binds to the KGF receptor, a splice variant of FGF receptor 2, which is expressed only in epithelial tissues. KGF has been identified as an important paracrine mediator of proliferation and differentiation in a wide variety of epithelial cells, including hepatocytes and gastrointestinal epithelial cells, type II pneumocytes, transitional urothelial cells, and keratinocytes in all stratified squamous epithelia. Systemic administration of recombinant human KGF (rHuKGF) provides significant cytoprotection to epithelial tissues in a number of different animal models of epithelial/mucosal damage, including models of injury to the gastrointestinal tract, lung, urinary bladder, and hair follicles. The results obtained with these preclinical models prompted an investigation of the use of rHuKGF as a cytoprotective agent against radiation- and/or chemotherapy induced oral and gastrointestinal mucositis. Several dose- and time-variable studies were conducted in normal rhesus macaques to determine the lowest dose and shortest duration of rHuKGF administration required to induce oral mucosal proliferation without other significant systemic effects. Numerous studies were also conducted in murine models of chemotherapy-induced mucositis to fine-tune the dosing schedule. These studies showed that 2-3 days of rHuKGF administration were sufficient to induce significant oral mucosal proliferation and to protect against gastrointestinal mucositis when administered prior to the initiation of chemotherapy. The results from these models were used to design a phase I study in normal human volunteers to evaluate the safety of rHuKGF and its ability to induce oral mucosal proliferation. rHuKGF was well tolerated and induced a significant increase in markers of oral mucosal proliferation following 3 days of administration at the highest doses. Phase I/II studies to evaluate the safety and efficacy of rHuKGF in the prevention of chemotherapy-induced mucositis are currently in progress. PMID- 10367677 TI - Development of a recombinant growth factor and fusion protein: lessons from GM CSF. AB - Several colony stimulating factors (CSFs) and cytokines have been successfully used to mobilize hematopoietic cells during myeloablative therapy, bone marrow failure, and transplantation and to provide supportive treatment during sepsis. The use of yeast-derived recombinant human granulocyte-macrophage CSF (rhuGM-CSF) and its interleukin-3 fusion protein, PIXY321, provides an example of issues associated with development programs for recombinant hematopoietic growth factors. Species specificity of rhuGM-CSF, different bioactivity of homologous molecules in mice, and production in laboratory animals of antibodies to human proteins limit preclinical evaluation of such molecules. In clinical trials, rhuGM-CSF was efficacious and well tolerated. The derivation of the recombinant molecule, optimal dosing, scheduling, and confounding effects of concurrent disease and treatments are factors that influence efficacy, adverse responses, and immunogenicity reported in patients treated with CSFs. In comparisons of yeast-derived with Escherichia coli-derived rhuGM-CS, the reduced severity and frequency of all adverse events, preponderance of low-grade adverse events, and similarity of positive clinical response versus adverse events reported for granulocyte CSF support safety and efficacy of yeast-derived rhuGM-CSE Enhanced pharmacoeconomic evaluations are beginning to limit and redirect clinical applications in this class of biological agents. PMID- 10367678 TI - Preclinical safety evaluation of rhuMAbVEGF, an antiangiogenic humanized monoclonal antibody. AB - Recombinant humanized antivascular endothelial growth factor (rhuMAbVEGF) is a monoclonal IgG1 antibody that is being developed as an antiangiogenic agent for use in treating a variety of solid tumors. Preclinical safety studies included an immunohistochemical tissue cross-reactivity study, in vitro hemolytic potential and blood compatibility studies, and multiple dose toxicity studies. Toxicity studies were conducted in cynomolgus monkey because rhuMAbVEGF is pharmacologically active in this species and does not bind rat or mouse vascular endothelial growth factor (VEGF). Following twice weekly administration of rhuMAbVEGF for 4 or 13 wk, young adult cynomolgus monkeys exhibited physeal dysplasia characterized by a dose-related increase in hypertrophied chondrocytes, subchondral bony plate formation, and inhibition of vascular invasion of the growth plate. In addition, decreased ovarian and uterine weights and an absence of corpora lutea were observed in females receiving 10 and 50 mg/kg/dose in the 13-wk study. Both the physeal and ovarian changes were reversible with cessation of treatment. No other treatment-related effects were observed following rhuMAbVEGF administration at doses up to 50 mg/kg. These findings indicate that VEGF is required for longitudinal bone growth and corpora lutea formation and that rhuMAbVEGF can reversibly inhibit physiologic neovascularization at these sites. PMID- 10367679 TI - BR96 sFv-PE40 immunotoxin: nonclinical safety assessment. AB - BR96 sFv-PE40, a recombinant DNA-derived fusion protein composed of the heavy- and light-chain variable region domains of the monoclonal antibody BR96 and the translocation and catalytic domains of Pseudomonas exotoxin A, is being developed for the treatment of solid tumors expressing cell surface Lewis(y)-related antigens. Single- and repeat-dose intravenous toxicity studies in rats and dogs and a comparative ex vivo tissue-binding study with rat, dog, and human tissues were conducted to assess the toxicity of BR96 sFv-PE40 and to estimate a safe starting dose in humans. Additional studies were performed to investigate the prevention of pulmonary vascular-leak syndrome, the dose-limiting toxicity of BR96 sFv-PE40 in rats, and the immunogenicity of BR96 sFv-PE40. In single-dose studies in rats, the vascular leak appeared to be primarily confined to the lungs; however, with a repeat-dose regimen (every other day for 5 doses) other organs including the brain and heart were involved at lethal doses (12-15 mg/m2 cumulative). Single doses of 1.8 mg/m2 and a cumulative 3.8 mg/m2 dose (0.75 mg/m2, every other day for 5 doses) were generally well tolerated in rats. These doses are significantly greater than doses required to cure rodents bearing human tumor xenografts. In dogs, the major target organ following single or repeated doses (every 3 days for 5 doses) was the pancreas. Morphologic changes in the exocrine pancreas ranged from atrophy with single-cell necrosis to diffuse acinar necrosis. After a 1-mo dose-free observation period, no residual pancreatic toxicity was observed in dogs given single doses up to 6.0 mg/m2 or 5 doses of 2.4 mg/m2 (12 mg/m2 cumulative). No significant pancreatic toxicity was observed at doses <0.6 mg/m2 in high Lewis(y)-expressing dogs. Assessment of trypsinlike immunoreactivity was useful in monitoring changes in pancreatic function. The immunogenicity of BR96 sFv-PE40 could be inhibited by combined treatment with an immunosuppressant in dogs, thus maintaining exposure to BR96 sFv-PE40. PMID- 10367680 TI - Correlation of toxicity and pharmacokinetic properties of a phosphorothioate oligonucleotide designed to inhibit ICAM-1. AB - ISIS 2302 is a phosphorothioate oligodeoxynucleotide with a sequence complementary to the mRNA of human intercellular adhesion molecule 1 (ICAM-1). Hybridization of ISIS 2302 to the mRNA inhibits expression of the ICAM-1 protein in response to inflammatory stimuli. A murine active antisense oligonucleotide, ISIS 3082, has been used for in vivo pharmacology studies and has anti inflammatory activity in models of organ transplant rejection, ulcerative colitis, and collagen-induced arthritis at doses ranging from 0.03 to 5 mg/kg. The safety assessment for ISIS 2302 includes general toxicity studies up to 6 mo in duration in mice and monkeys, genetic toxicity studies, and reproductive/fertility studies. ISIS 3082 was examined in parallel with ISIS 2302 in mouse toxicity and reproductive studies. The toxicities observed following systemic administration of ISIS 2302 and ISIS 3082 were similar and consistent with those observed for other compounds in this chemical class and, therefore, are independent of the suppression of ICAM-1 expression. Toxicokinetic evaluation demonstrated that toxicities occurred in organs containing the highest concentrations of ISIS 2302. Evidence of immune stimulation. including dose dependent splenomegaly, lymphoid hyperplasia, and multiorgan mixed mononuclear cell infiltrates, was the most common finding in rodent studies. Monkeys were much less sensitive than mice to immune stimulation. Kidney contained the highest concentrations of ISIS 2302. Morphologic changes observed in kidney included atrophic and regenerative changes in proximal tubular epithelium; however, there was no evidence of functional abnormalities. Additional histologic changes noted in proximal tubular epithelium included basophilic granules, which were reflective of oligonucleotide distribution and uptake in these cells. Liver also contained high concentrations of oligonucleotide, which were associated with Kupffer cell hypertrophy in mice. Changes in serum transaminases, cholesterol, and triglycerides were reflective of hepatic alterations. In monkeys, high concentrations of oligonucleotide caused a transient increase in clotting times and activation of the alternative complement pathway. All toxicities associated with ISIS 2302 were reversible and occurred at doses well above those required for pharmacologic activity or currently used in clinical trials. In addition, there has been no evidence of genetic toxicity associated with ISIS 2302, and no changes in reproductive performance, fertility, or fetal development have been noted in animals treated with ISIS 2302 or ISIS 3082. PMID- 10367681 TI - Safety evaluation of human living skin equivalents. AB - Human living skin equivalents (LSEs) offer an alternative to the use of split thickness autografts for the treatment of hard-to-heal wounds. LSEs consist of 4 active components: a well-differentiated stratum corneum derived from epidermal keratinocytes, dermal fibroblasts, and an extracellular collagen matrix. Neonatal foreskins are used as the source of keratinocytes and dermal fibroblasts for the manufacture of LSEs. Following isolation and expansion in vitro, the cells are cultured on a 3-dimensional scaffold to give an upper epidermal layer and supporting dermal layer. The resulting product has the appearance and handling characteristics of human skin. Safety evaluation of LSEs begins with insuring that foreskins are obtained only from healthy infants whose mothers are negative for a panel of adventitious agents. Keratinocyte and fibroblast cell banks are characterized using morphologic, biochemical, and histologic criteria; checked for the absence of contaminating cell types such as melanocytes, macrophages, lymphocytes, and Langerhans cells; subjected to rigorous microbiological testing (with any production materials of biological origin); and evaluated for in vivo tumorigenicity. The consistency of certain key morphologic and functional characteristics are regularly assessed. Because an LSE represents an allogeneic graft, preclinical safety studies include in vitro and in vivo determinations of its potential immunogenicity. Immunocompromised (SCID) mice reconstituted with human leukocytes or engrafted with human fetal hematolymphoid organs have been useful animal models for assessing possible immunologic responses to LSEs. Additional preclinical studies are being conducted to show that LSEs are noncytotoxic and lack allergenic, sensitizing, or irritation potential. PMID- 10367682 TI - The pathologist and toxicologist in pharmaceutical product discovery. AB - Significant change is occurring in the drug discovery paradigm; many companies are utilizing dedicated groups from the toxicology/ pathology disciplines to support early stage activities. The goal is to improve the efficiency of the discovery process for selecting a successful clinical candidate. Toxicity can be predicted by leveraging molecular techniques via rapid high-throughput, low resource in vitro and in vivo test systems. Several important activities help create a platform to support rapid development of a new molecular entity. The proceedings of this symposium provide excellent examples of these applied concepts in pharmaceutical research and development. Leading biopharmaceutical companies recognize that a competitive advantage can be maintained via rapid characterization of animal models, the cellular identification of therapeutic targets, and improved sensitivity of efficacy assessment. The participation of the molecular pathologist in this quest is evolving rapidly, as evidenced by the growing number of pathologists that interact with drug discovery organizations. PMID- 10367683 TI - Revolution through genomics in investigative and discovery toxicology. AB - The remarkable technologic and methodologic advances spurred on by the Human Genome Project are being applied throughout the life sciences. In the field of toxicology, high-resolution assays now make it possible to discover virtually all the differences in gene expression brought on by exposure to a particular xenobiotic. There are 2 principal approaches used to build a catalog of changes in gene expression: hybridization microarrays and gel-based methods, such as differential display and AFLP-based mRNA finger-printing. The power of such approaches is exemplified by the identification of more than 300 genes that differ in expression level by at least 2-fold in response to the nongenotoxic rodent liver carcinogen phenobarbital. PMID- 10367684 TI - Utilization of genetically altered animals in the pharmaceutical industry. AB - The study of transgenic and gene-deleted (knockout) mice provides important insights into the in vivo function and interaction of specific gene products. Within the pharmaceutical industry, genetically altered mice are used predominantly in discovery research to characterize the diverse functions of one or multiple gene products or to establish animal models of human disease for proof-of-concept studies. We recently used genetically altered animals in drug discovery to examine the NF-kappaB family of transcriptional regulatory genes and to elucidate their essential role in the early onset of immune and inflammatory responses. Transgenic and knockout mice are also useful in drug development, because questions regarding risk assessment and carcinogenesis, xenobiotic metabolism, receptor- and ligand-mediated toxicity, and immunotoxicity can be evaluated using these genetically altered mice. For example, the p53 knockout mouse is one of several genetically altered mice whose use may increase the sensitivity and decrease the time and cost of rodent carcinogenicity bioassays. As with any experimental model system, data obtained from genetically altered mice must be interpreted carefully. The complete inactivation of a gene may result in altered expression of related genes or physiologic compensation for the loss of the gene product. Consideration must also be given to the genetic background of the mouse strain and the impact of strain variability on disease or toxicity models. Despite these potential limitations, knockout mice provide a powerful tool for the advancement of drugs in the pharmaceutical industry. PMID- 10367685 TI - Mechanisms of disease and injury: utilization of mutants, monoclonals, and molecular methods. AB - Rapid advances in our ability to localize and quantify macromolecular changes in health and disease are being brought about by the availability of genetically altered animals (mutants), purified reagents such as monoclonal antibodies, and new molecular methods. Targeted gene deletion (knockouts) and gene insertions (transgenics) in animals can allow identification of the importance and function of macromolecules. Monoclonal antibodies and fluorescent labels coupled with advances in microscopy provide exacting and multi-dimensional information about localization and cellular changes in proteins, carbohydrates, and lipids using immunohistochemistry, fluorescent activated cell sorting, and immunoprecipitation. Similarly, new applications of molecular methods can be used to identify and localize nucleic acids in tissues via in situ hybridization, polymerase chain reaction (PCR), reverse transcription (RT) PCR, differential display RT-PCR, RNase protection assays, and microchip arrays. The ligand for CD40 (CD40L), an important immunoregulatory molecule, is an example of the successful application of mutants, monoclonal antibodies, and molecular methods to cloning and biological characterization of new molecules. CD40L knockout mice, monoclonal antibodies, and several molecular methods were used to identify mutations in CD40L as the genetic basis for hyper-IgM syndrome in humans, to provide new insights into the pathobiology of Pneumocystis carinii infection, and to evaluate CD40L for immunotherapy of tumors and opportunistic infections. PMID- 10367686 TI - Tumor models: assessing toxicity in efficacy studies. AB - Efficacy studies in animal tumor models provide an early opportunity to collect preliminary information on toxicity. When screening and evaluating cytotoxic chemotherapeutic agents, efficacy studies usually include at least 1 dosage level that causes severe toxicity and death. Pathologic evaluation in early efficacy studies may reveal major target organs, dosage/schedule relationships, pharmacokinetic/toxicity relationships, effects of formulation and route of administration, maximum tolerated dose, cause of death, and reversibility of changes in normal tissues. Intraperitoneal formulations are frequently used to establish proof of concept for promising compounds (hits) from in vitro screens; however, these crude formulations may also induce intraperitoneal inflammation and confound the interpretation of both efficacy and toxicity. Efficacy studies conducted in the later stages of drug discovery may be used to refine the dose and schedule proposed for phase I clinical trials. Efficacy studies in animal tumor models provide useful toxicologic data for screening potential drug candidates, optimizing the therapeutic index, and designing both preclinical and clinical development programs. PMID- 10367687 TI - The role of IL-10 in inflammatory bowel disease: "of mice and men". AB - Inflammatory bowel disease (IBD) is a generic term typically used to describe a group of idiopathic inflammatory intestinal conditions in humans that are generally divided into Crohn's disease and ulcerative colitis. Although the etiology of these diseases remains unknown, a number of rodent models of IBD have recently been identified, all sharing the concept that the development of chronic intestinal inflammation occurs as a consequence of alterations in the immune system that lead to a failure of normal immunoregulation in the intestine. On the basis of these models, it has been hypothesized that the development of IBD in humans may be related to a dysregulated immune response to normal flora in the gut. Immunodeficient scid mice injected with CD4+ CD45RB(high) T cells and mice deficient in interleukin (IL)-10 (IL-10-/-) are among the rodent models of IBD. In both models, there is inflammation and evidence of a Th1-like response in the large intestine, characterized by CD4+ T-cell and macrophage infiltrates, and elevated levels of interferon-gamma. Because IL-10 is an immunomodulatory cytokine that is capable of controlling Th1-like responses, the role of IL-10 was investigated in these models. IL-10 was shown to be important in regulating the development of intestinal inflammation in both models. These results provided key data that supported initiation of clinical trials evaluating the efficacy of IL 10 in patients with IBD. PMID- 10367688 TI - Animal models of arthritis: relevance to human disease. AB - Animal models of arthritis are used to evaluate potential antiarthritis drugs for clinical use. Therefore capacity of the model to predict efficacy in human disease is one of the most important criteria in model selection. Animal models of rheumatoid arthritis (RA) with a proven track record of predictability include rat adjuvant arthritis, rat type II collagen arthritis, mouse type II collagen arthritis, and antigen-induced arthritis in several species. Agents currently in clinical use (or trials) that are active in these models include corticosteroids, methotrexate, nonsteroidal anti-inflammatory drugs, cyclosporin A, leflunomide, interleukin-1 receptor antagonist, and soluble tumor necrosis factor receptors. For some of these agents, the models also predict that toxicities seen at higher doses for prolonged periods would preclude dosing in humans at levels that might provide disease-modifying effects. Animal models of osteoarthritis (OA) include mouse and guinea pig spontaneous OA, meniscectomy and ligament transection in guinea pigs, meniscectomy in rabbits, and meniscectomy and cruciate transection in dogs. None of these models have a proven track record of predictability in human disease because there are no agents that have been proven to provide anything other than symptomatic relief in human OA. Efficacy data and features of the various models of RA and OA are discussed with emphasis on their proven relevance to human disease. PMID- 10367689 TI - Preclinical development of agents for the treatment of osteoporosis. AB - Because of the high cost and long time frame of clinical testing, animal models play a crucial role in the identification and selection of agents for the treatment of osteoporosis. The use of animal models early in a program focuses on the establishment of efficacy, while animal models used later in a program to examine bone safety. More specifically, animal models are used to gain information on the skeletal mechanism of action, to examine multiple skeletal sites (axial and appendicular), and to examine the effects of higher doses than will be used in humans. Animal models also predict the usefulness of surrogate markers in clinical trials, such as formation and resorption markers, as well as bone density. The hazard of using surrogate markers for fracture prevention is highlighted by high dose fluoride administration, which can increase bone density (considered a strong predictor of fracture protection) while not protecting against fractures. Estrogen-deficient models are most commonly used to mimic the postmenopausal bone loss in women; these models are characterized by increased bone turnover and a negative bone balance. The timing of the administration of the new therapy in animal models can help determine whether the agent will be more effective in the prevention of osteoporosis or in the treatment of established osteoporosis. New methods for the measurement of bone mass or volume are less invasive, require shorter acquisition time, and have enhanced resolution, resulting in increased knowledge concerning architectural changes and specific sites of bone deposition. Finally, the measurement of biomechanical strength of bones from animal models can be used to predict protective effects on fracture rates in clinical trials. When used in combination with other methods, animal models can greatly increase our understanding of the pathophysiology of osteoporosis and can expedite the development of new therapies. PMID- 10367690 TI - Opportunistic infections of the central nervous system during HIV-1 infection (emphasis on cytomegalovirus disease). AB - Toxoplasma encephalitis, cryptococcal meningitis, progressive multifocal leukoencephalopathy (PML), and cytomegalovirus (CMV) encephalitis are the most common opportunistic infections of the central nervous system (CNS) in HIV infected patients. They occur at variable degrees of immunosuppression, and their diagnosis is based on a systematic evaluation with includes, in a definite order, ongoing prophylactic therapies, extraneurological signs, neuroimaging and CSF studies, and an anti-Toxoplasma therapeutic trial. Concurrent neurological HIV CNS disease (such as the AIDS dementia complex) is frequent. The development of reliable molecular biology techniques such as the polymerase chain reaction and their application to the CSF have made the diagnosis of virus-related opportunistic infections much easier and has limited the need for cerebral biopsy. The incidence of opportunistic infections has decreased since the introduction of recent antiretroviral therapeutic strategies. PMID- 10367691 TI - Endovascular treatment of ruptured intracranial aneurysms. AB - The Guglielmi detachable coil (GDC) is an important tool for the treatment of ruptured intracranial aneuryms by an endovascular approach. This new device may be introduced under fluoroscopy into the aneurysmal sac through a microcatheter. When the coil is judged well positioned, it can be detached with accuracy by electrolytic breakdown. The procedure is completed when a dense coil packing is obtained. When vasospasm is present, papaverin infusion or angioplasty may be used by the endovascular approach as well. Best results are achieved in cases of small aneurysm with small neck. The morbidity and mortality rates in the first 200 patients treated by GDC for a ruptured intracranial aneurysm were 4% and 1.5%, respectively. Complications are generally related to rupture of the malformation by the endovascular device or to thromboembolic events. Despite these promising results, further studies using larger numbers of patients are required to determine the exact role of these procedures in patient care. PMID- 10367692 TI - The role of the intact hemisphere in recovery of midline muscles after recent monohemispheric stroke. AB - Transcranial magnetic stimulation (TMS) of the motor cortex was used to study basic mechanisms of motor reorganization after major hemispheric stroke in humans. We sought to clarify the possible role of the intact hemisphere in motor recovery of the lingual muscles, and to evaluate the compensatory use of preexisting uncrossed motor pathways projecting to these midline muscles. TMS and bilateral surface recordings from the lingual muscles were carried out in six selected stroke patients who presented with a unilateral lingual paralysis after a limited monohemispheric ischemia. The first examination was performed during the symptomatic stage (t1) and was repeated after complete recovery of lingual function had been established (t2). The cortical motor output patterns were analyzed and compared with the data from 40 healthy controls. In the controls TMS of either hemisphere invariably produced contralateral and ipsilateral compound muscle action potentials (CMAPs), elicited through crossed and uncrossed central motor pathways, respectively. In most individuals an asymmetric cortical motor output pattern was found, as significantly greater mean CMAPs of shorter onset latencies were recorded from the contralateral lingual muscles than from the ipsitateral responses. In the six patients with a unilateral lingual paralysis a similar pattern was found on initial examination by stimulating the intact hemisphere, whereas TMS of the affected hemisphere failed to elicit any CMAP bilaterally. At t2 all patients had regained normal lingual function. Only one patient showed evidence of a complete recovery of the primarily affected hemisphere, as TMS now elicited normal CMAPs bilaterally. In the remaining five patients the unilateral interruption of the corticonuclear pathways persisted in spite of complete functional recovery. In these subjects the recovery of symmetric lingual movements must be attributed to the intact hemisphere. From this it is concluded that recovery of a unilateral lingual paralysis after restricted monohemispheric lesions is possible without recovery of the cortical motor projections from the affected hemisphere. In these cases the intact hemisphere is responsible for restoration of normal lingual movements, most likely by potentiating the effect of preexisting uncrossed motor pathways. PMID- 10367693 TI - Surgical and medical management of patients with massive cerebellar infarctions: results of the German-Austrian Cerebellar Infarction Study. AB - Surgical intervention (ventricular drainage or decompressive craniotomy) may be necessary in patients with cerebellar infarction if mass effect develops. However, patient selection and timing of surgery remain controversial, and there are few data on clinical signs in the early course that are predictive for outcome. The clinical course and neuroradiological features of 84 patients (aged 22-78, mean 58.5 years) with massive cerebellar infarction confirmed by computed tomography were prospectively observed for 21 days after admission and at 3-month follow-up using a standardized protocol. Data were gathered from 1992 to 1996 in 17 centers. The patients were assigned to three treatment groups depending on the decision of the primary caretaker: 34 underwent craniotomy and evacuation, 14 received ventriculostomy, and 36 were treated medically. Treatment groups differed regarding the level of consciousness, signs of mass effect in computed tomography and signs of brainstem involvement. The overall risk for poor outcome depended on the level of consciousness after clinical deterioration (odds ratio = 2.8). Subgroup analysis of awake/drowsy or somnolent/stupor patients revealed no relationship to treatment. The vascular territory involved did not affect outcome. Surgical treatment for massive cerebellar infarctions was not found to be superior to medical treatment in awake/drowsy or somnolent/stupor patients. Half of all patients deteriorating to coma treated with ventricular drainage or decompressive craniotomy had a meaningful recovery. We were unable to compare surgical versus medical therapy in this subgroup due to lack of control group. This study supports the notion that the level of consciousness is the most powerful predictor of outcome, superior to any other clinical sign and treatment assignment. Deterioration of consciousness typically occurred between days 2 and 4, with a maximum on day 3. PMID- 10367694 TI - Long-term treatment of cervical dystonia with botulinum toxin A: efficacy, safety, and antibody frequency. German Dystonia Study Group. AB - Data from 616 patients suffering from idiopathic cervical dystonia were analyzed to determine the efficacy and safety of treatment with botulinum neurotoxin type A (BoNT/A). Since the specific purpose of this study was to determine the long term effects of this treatment, the analysis focused specifically on the patients (n = 303) having received six or more injections. Statistical analysis of a standardized documentation showed sustained significant benefit as measured by a disease severity score independent of the type of cervical dystonia. Furthermore, pronounced individual differences were found in response to this treatment although initial clinical scores and doses of BoNT/A were similar. There was no indication of previously unknown adverse events, the only risk being the development of serum antibodies against the toxin. As in previous studies on short-term effects of BoNT/A treatment, the most frequent adverse event was dysphagia, which occurred on average 9.7 days after injection and lasted on average 3.5 weeks. While secondary nonresponse was seen in approx. 5% of patients, antibody tests revealed neutralizing serum antibodies in only 2%. On the basis of the present data, therapy of cervical dystonia with BoNT/A seems to be safe and yields good stable results even after 5 years of treatment. PMID- 10367695 TI - Daytime somnolence in myotonic dystrophy. AB - Somnolence in myotonic dystrophy (DM) has not been measured using a reliable daytime somnolence scale. The aim of this study was to compare somnolence in DM patients with healthy controls and Charcot-Marie-Tooth disease (CMT) patients using such a scale and to compare this with potential contributory factors. We investigated 35 subjects with adult-onset DM, 16 healthy controls and 13 CMT controls. The Epworth Sleepiness Scale (ESS) was the principal measurement of daytime somnolence. Nocturnal sleep was assessed using a sleep diary. Other assessments measured daytime respiratory function, cognitive function, motor impairment, disability, swallowing capacity and depression. DM and CMT patients had greater daytime sleepiness than unaffected controls. In the DM group significant correlations were found between somnolence and measures of disability, sleep quality and some measures of depression. It was concluded that there is an abnormal level of daytime somnolence in DM, which is partially associated with disability. PMID- 10367696 TI - Idiopathic blepharospasm does not lead to a parkinsonian syndrome: results of a questionnaire-based follow-up study. AB - It has been suggested that a lesion in the dopaminergic neurons of the substantia nigra pars compacta combined with eye irritation is involved in the pathophysiology of idiopathic blepharospasm. If so, these patients might be prone to develop Parkinson's disease or a parkinsonian syndrome. We therefore conducted a validated questionnaire-based follow-up study to estimate (a) the frequency of local eye disorders at onset and (b) frequency of development of parkinsonian symptoms in blepharospasm patients. Ninety-nine patients previously diagnosed with idiopathic blepharospasm were sent a two-part questionnaire to assess parkinsonian and other symptoms associated with their condition. The average period of follow-up was 12.7 years, ranging from 3 to 26 years, with an average age at onset of 53.5 years. Sixty-two patients reported other ocular symptoms prior to or at the onset of blepharospasm, and therefore ocular disease may act as a trigger to produce blepharospasm in those already predisposed. Only two patients had developed a score on the parkinsonism rating scale indicating possible Parkinson's disease, but clinical examination confirmed this not to be the case. If a lesion in the dopaminergic neurons is involved in blepharospasm, it would appear to be relatively minor (and non-progressive), since patients with idiopathic blepharospasm do not seem prone to develop parkinsonian symptoms. PMID- 10367697 TI - Spontaneous primary intraventricular hemorrhage: clinical data, etiology and outcome. AB - The clinical features, etiology, and neurological outcome in patients with primary intraventricular hemorrhage (PIVH) have rarely been reported. We retrospectively reviewed the clinical data, complementary examinations, outcome, computed tomography (CT) blood amount, and ventricle size of 13 patients (mean age 60 years, five men). We defined PIVH as hemorrhage detected by CT in the ventricular system only. The major symptoms included headache (n = 13), decreased level of consciousness (n = 9), and nausea/vomiting (n = 7). The cause was unknown in five patients; and was associated with arterial hypertension in five, vascular malformations in two, and tumor in one, although arteriography was performed in only five patients. Outcomes were death in three, asymptomatic in six, mild disability in three, and moderate disability in one. Prognosis was not related to clinical or CT data. Clinical features can suggest the diagnosis of PIVH, but cerebral CT is required for confirmation. PMID- 10367698 TI - Binding characteristics of the glucocorticoid receptor in peripheral blood lymphocytes in multiple sclerosis. AB - Although the exact etiology of multiple sclerosis (MS) remains unresolved, immune reactions are believed to be the central pathogenic mechanisms. Endogenous and therapeutic steroid hormones affect the immune system, and inflammatory diseases are associated with activation of the hypothalamic-pituitary-adrenal axis, providing evidence of an immune-endocrine interplay. Function tests in MS have revealed dysregulation of the hypothalamic-pituitary-adrenal system in a substantial proportion of patients. We characterized glucocorticoid receptor (GR) binding in peripheral blood lymphocytes from 39 MS patients and 14 age- and sex matched controls with respect to dissociation constant and binding capacity, using a whole-cell binding assay with [3H]dexamethasone as the ligand. GR binding parameters did not differ significantly between patients (Kd 8.98 +/- 1.07 nM, Bmax 183 +/- 29.8 fmol/mg) and controls (Kd 9.36 +/- 1.17 nM, Bmax 158 +/- 16 fmol/mg). No effect of age, sex, course, duration or severity of disease, or prior steroid treatments was detected. GR binding parameters were analyzed in relation to the results of the combined dexamethasone-CRH test, which reflects corticosteroid receptor function at the hypothalamus, in 30 patients and 9 controls. While controls showed a moderate correlation between binding affinity of the GR in lymphocytes and regulatory function at the hypothalamic level, the patients did not. These data suggest that the physiological relationship between binding and function of the glucocorticoid receptor is disturbed in MS. PMID- 10367699 TI - Intravenous immunoglobulins in the therapy of paraneoplastic neurological disorders. AB - The treatment of paraneoplastic neurological syndromes (e.g., tumor therapy, immunosuppressive therapy, plasmapheresis) rarely leads to an improvement in the neurological symptoms. We treated four patients suffering from paraneoplastic neurological syndromes with intravenous immunoglobulins. All four had high titers of antineuronal antibodies in serum and CSF. Two of the patients, one suffering from paraneoplastic cerebellar degeneration and the other from paraneoplastic brain stem encephalitis and polyneuropathy, received intravenous immunoglobulin treatment within 3 weeks of the onset of neurological symptoms. Both patients showed clinical improvement within 2 weeks after the initiation of therapy. They also showed a decline in the intrathecal antibody synthesis of the antineuronal antibody. Two other patients, who had suffered from paraneoplastic neuropathy for 3 and 6 months showed no improvement with the intravenous immunoglobulin therapy. In these cases there was no effect on intrathecal antibody synthesis. When started early, intravenous immunoglobulins may be of therapeutical value in treating paraneoplastic neurological syndromes. Specific intrathecal antibody synthesis may be a better measure of clinical course that autoantibody serum titers. PMID- 10367700 TI - Apolipoprotein E genotype and progression of Alzheimer's disease: the Rotterdam Study. AB - The APOE*4 allele of the apolipoprotein E gene increases the risk of Alzheimer's disease (AD), but whether it also affects the course of the disease is controversial. However, all studies on this issue until now have been based on patients at various stages of disease. In the present population-based study, 97 patients were included at a similar stage, i.e., before the onset of symptoms, and followed for up to 5 years. We found that the APOE*4 allele is not a strong determinant of survival in AD. As change in cognitive function and severity of dementia are similar for AD patients with and without APOE*4, our study suggests that progression of AD is not related to the APOE*4 allele. PMID- 10367701 TI - Deterioration in parkinsonism with low-dose pergolide. AB - The administration of dopamine agonists can have a role early in the course of Parkinson's disease, in an attempt to reduce the frequency of long-term motor complications associated with the use of levodopa. After treatment with dopamine agonists has begun, gradual dose escalation is recommended to reduce the incidence of side effects; at low doses, parkinsonian symptoms significantly decline in some patients, only to improve as the dose increases. We report a number of such patients and discuss the possible pathogenesis of this motor deterioration. PMID- 10367702 TI - Cerebellar spatial dysgraphia: further evidence. PMID- 10367703 TI - Non-communicating syringomyelia and neuromyelitis optica. PMID- 10367704 TI - Cognitive profile in dementia associated with vitamin B12 deficiency due to pernicious anaemia. PMID- 10367705 TI - Systemic sarcoidosis: a case with a focal hydrocephalus and elevated lysozyme and angiotensin-converting enzyme in the cerebrospinal fluid. PMID- 10367706 TI - Nickel-induced activation of T cells in individuals with negative patch test to nickel sulphate. AB - Contact hypersensitivity to nickel is the most common form of allergic contact dermatitis. To gain insight into the induction of this frequent disease, T cell reactivity towards nickel was investigated in "nonallergic" individuals defined as those with no skin manifestations and a negative patch test towards NiSO4. Surprisingly, we found that nickel induced proliferation of peripheral blood mononuclear cells (PBMC) from 16 of 18 adult individuals tested. This activation was specific, and no stimulation of PBMC was observed using control stimulants at equimolar concentrations. Furthermore, the NiSO4-induced activation required the presence of professional antigen-presenting cells. To describe the functional capacity of the nickel-inducible T cells, cytokine release was investigated in both nickel-allergic and nonallergic individuals. The T cells from both groups released interferon-gamma but no interleukin-4 upon stimulation with nickel, suggesting that the functional capacities of these cell populations were similar in nickel-allergic and nonallergic individuals. Thus, at this level, no qualitative differences could be demonstrated between T cells obtained from nickel-allergic and nonallergic individuals. PMID- 10367707 TI - Basal cell carcinoma possibly originates from the outer root sheath and/or the bulge region of the vellus hair follicle. AB - In the present study, the immunophenotype of basal cell carcinoma was analysed in comparison with human vellus hair follicular keratinocytes. We also established the lectin binding profile of basal cell carcinoma and human vellus hair follicles (VHF), using several lectins with different sugar specificities. Our findings showed an almost identical immunohistochemical profile for basal cell carcinoma and the suprabulbar region of the outer root sheath of VHF, whereas other follicular compartments such as the bulbar, the isthmus or the supraseboglandular regions did not correlate. In particular, homogeneous and constant expression of the basal differentiation markers CK 5 and CK 14 were found in both specimen, with no expression of the simple epithelium type keratin CK 8 and the suprabasal differentiation markers CK 1 and CK 10. CK 19 showed variable expression in basal cell carcinoma, with constant expression in the outer root sheath and the follicular bulge regions, but was always absent in interfollicular epidermal keratinocytes. In addition, the lectin binding profiles of basal cell carcinoma and the outer root sheath in the suprabulbar region of human VHF were comparable, with the presence of binding sites for PNA, Con A and WGA. These findings provide evidence for a histochemical relationship between basal cell carcinoma and the follicular epithelium of VHF which is closer than that with the epidermis, and suggest its possible origin from or possibly its differentiation pattern towards the cells of the outer root sheath and/or the follicular bulge region of the VHF. PMID- 10367708 TI - Stratum corneum thiol protease (SCTP): a novel cysteine protease of late epidermal differentiation. AB - Proteolytic enzymes play crucial roles in the formation of the stratum corneum barrier tissue and in its subsequent maturation. Despite this, the proteases involved in stratum corneum physiology are not well characterized. Hence, studies were performed to identify these proteolytic enzymes present in the peripheral layers of this tissue using a combination of tape stripping and zymography. Using this approach, a novel human cysteine protease was identified and characterized, and named stratum corneum thiol protease (SCTP). Gelatin zymography revealed that SCTP is composed of two variants with apparent molecular weights of 34 and 35 kDa which do not correspond to any previously described stratum corneum protease. Mechanistically SCTP belongs to the cysteine proteinase class as shown by: (1) acid protease activity, (2) a requirement for mild reducing conditions, and (3) the specific inhibition of activity by E64 and Z-phe-ala-diazomethylketone. Further analysis using concanavalin A affinity chromatography demonstrated that the two 34 and 35 kDa variants are both glycoproteins, which, after removal of the oligosaccharide sidechains with the specific enzyme N-glycopeptidase F, reveal a single active core protease of 32 kDa. SCTP did not crossreact with antibodies raised against the lysosomal cysteine proteases cathepsins B, H or L, thereby distinguishing it from the classical cysteine cathepsins. Localization studies revealed that SCTP is present at all depths in the stratum corneum, thereby precluding microbial contamination as the enzyme source. Moreover, it was also present at all body sites investigated, except for the hyperkeratotic palmoplantar stratum corneum. SCTP was found to be a product of late differentiation in cultured human keratinocytes; the enzyme was synthesized by differentiated calcium-switched cells and secreted into the medium, whereas nondifferentiated basal keratinocytes did not produce this protease. Moreover, human fibroblast cultures did not produce the enzyme, suggesting that SCTP is not produced by the dermis and hence is epidermal specific. The function of SCTP is unknown, but the observed gelatinolytic activity coupled with its secretion into the medium by cultured keratinocytes indicates that physiologically it is responsible for the degradation of extracellular structural proteins. Furthermore, the optimal activity at acid pH suggests that it can function in the acidic environment of the stratum corneum. It remains to be elucidated whether this enzyme has a role in desquamation. PMID- 10367709 TI - Serotonin in human allergic contact dermatitis. An immunohistochemical and high performance liquid chromatographic study. AB - Allergic contact dermatitis (ACD) is a common clinical condition leading to considerable morbidity. We have recently demonstrated that ketanserin, a serotonin antagonist, significantly inhibits nickel sulphate-induced ACD. Furthermore, serotonin-immunoreactive (IR) cells have previously been demonstrated in normal human cutaneous melanocytes. To further elucidate the role of serotonin in cutaneous contact hypersensitivity, we compared ACD involved skin and uninvolved skin from nickel-allergic patients, and normal skin from healthy volunteers, for the presence of serotonin-like immunoreactive cells using immunohistochemistry. In addition, serotonin concentrations in ACD involved and uninvolved skin were compared by high-performance liquid chromatography (HPLC). In the skin of normal healthy volunteers, the serotonin-IR cells were situated in the basal layer of the epidermis. In uninvolved skin the cells were also situated in the basal layer, but they were more numerous and the immunofluorescence intensity was greater. In involved skin, the IR cells were fewer and they were found higher up in the epidermis. Also, the configuration of these cells was different: they showed enlarged and elongated dendrites as well as dendritic spines. The serotonin antiserum-labelled cells in ACD involved skin were also NKI beteb positive (the latter is known as a reliable marker of melanocytes). The concentration of serotonin in involved skin was significantly higher than that in uninvolved skin in ACD patients (P < 0.05). Taken together, our previous and present results indicate that serotonin plays an important role in ACD. The basal epidermal serotonin-IR cells are more dendritic in ACD, and are found more superficial in the epidermis, where they might release their content of serotonin, thereby influencing the inflammatory process. PMID- 10367710 TI - FK506 and cyclosporin A inhibit stem cell factor-dependent cell proliferation/survival, while inducing upregulation of c-kit expression in cells of the mast cell line MC/9. AB - Murine mast cell proliferation and maturation are regulated by two distinct cytokines, interleukin-3 (IL-3) and the c-kit ligand, stem cell factor (SCF). In this study using cells of the mouse mast cell line, MC/9, the effects of two immunosuppressants, FK506 and cyclosporin A (CsA), were investigated. Withdrawal of IL-3 from the culture medium resulted in loss of viability of MC/9 cells. The addition of SCF in the absence of IL-3 maintained MC/9 cell survival but no cell proliferation was detected. The combined addition of IL-3 and SCF to the culture medium resulted in a more marked MC/9 cell proliferation than the addition of IL 3 alone. FK506 and CsA inhibited the SCF-dependent, but not the IL-3 dependent, stimulatory effects on MC/9 cell proliferation/survival. Apoptotic changes were analyzed using fluorescent staining with acridine orange and DNA electrophoresis. FK506 and CsA inhibited the SCF-dependent rescue effect from apoptosis. Flow cytometry showed that FK506 and CsA did not affect IL-3 receptor expression. However, immunoblot and reverse transcriptase-polymerase chain reaction (RT-PCR) analyses indicated that c-kit protein and c-kit mRNA transcripts were increased following the FK506 and CsA treatments in the presence of IL-3. In addition, MC/9 cells pretreated with FK506 or CsA showed an increased adhesiveness to NIH/3T3 cells that express membrane-bound SCF. Neither FK506 nor CsA affected c-kit tyrosine phosphorylation or MAP kinase nuclear translocation of MC/9 cells following SCF stimulation. These results indicate that FK506 and CsA, while inducing c-kit of MC/9 cells, selectively inhibit the SCF-dependent stimulatory effects on MC/9 cell proliferation/survival by a mechanism independent of, or at point(s) distal to, the c-kit-MAP kinase pathway. PMID- 10367711 TI - Ceramide potentiates, but sphingomyelin inhibits, vitamin D-induced keratinocyte differentiation: comparison between keratinocytes and HL-60 cells. AB - Differentiation of epidermal keratinocytes and leukemia HL-60 cells induced by 1,25-dihydroxyvitamin D [1,25(OH)2D] has been reported to be mediated, at least in part, by increases in cellular ceramide levels. Ceramides produced by 1,25(OH)2D-induced sphingomyelin (SM) hydrolysis also contribute to the permeability barrier lipids in keratinocytes. Exogenously supplied SM is taken up by mammalian cells, including keratinocytes, and is incorporated into cellular pools. However, the effects of exogenously added SM on keratinocyte differentiation have not been studied. Therefore, in this study, we compared exogenously added SM with a cell-permeable ceramide for their ability to stimulate keratinocyte differentiation induced by 1,25(OH)2D. Both short-chain ceramide (C2-cer) and SM stimulated the differentiation and inhibited the proliferation of HL-60 cells. As expected, this effect was potentiated by 1,25(OH)2D. However, SM inhibited the differentiation and stimulated the proliferation of keratinocytes. While C2-cer potentiated the effects of 1,25(OH)2D, SM reversed the effects of 1,25(OH)2D on keratinocytes. The ratio of SM to ceramide was significantly different between keratinocytes and HL-60 cells. While the SM level of HL-60 cells were twice that of keratinocytes, keratinocytes contained ten times more ceramides than HL-60 cells, resulting in a ceramide/SM ratio 17 times higher in keratinocytes. Thus, we identified similarities and significant differences in the sphingolipid-mediated cell signaling pathway between keratinocytes and HL-60 cells. While SM stimulated HL-60 cell differentiation, presumably by incorporation into SMase-accessible membrane pools, it inhibited keratinocyte differentiation. In keratinocytes, SM was possibly incorporated into a different cellular pool (barrier lipid pool) or altered membrane phospholipid metabolism and membrane fluidity. PMID- 10367712 TI - Characterization of the chick chorioallantoic membrane model as a short-term in vivo system for human skin. AB - We report on the cultivation and characterization of human skin on the chorioallantoic membrane of chicken eggs with the aim of replacing animals in short-term investigations in dermatology. Adult human split-thickness skin was grafted onto the chorioallantoic membrane of 5-day chick embryos. Grafts and surrounding host tissue were examined daily by in vivo stereomicroscopy and in histological sections and were characterized using a panel of monoclonal antibodies. The skin grafts were completely incorporated into the chorioallantoic membrane 2 days after transplantation. A remarkable angiogenesis occurred towards the grafts. Skin tissues revascularized within 2 or 3 days by reperfusion of the existing graft vasculature. Anastomosis of host and graft blood vessels occurred and the transplanted skin was nourished by the host blood supply as indicated by nucleated chick erythrocytes in the skin vessels. The skin grafts on the chorioallantoic membrane preserved an almost entire human phenotype. Besides a fully differentiated human epidermis and dermis containing all the cellular and extracellular constituents such as skin immune cells, capillary vessels composed of human endothelial cells were enclosed by a basement membrane of human origin. The integrin expression pattern formed in human skin transplants 5 days after grafting was identical to that of human skin controls before grafting. PMID- 10367713 TI - Topical amikacin formulation induces fibroblast growth factor and cytokine release from human dermal fibroblasts. PMID- 10367714 TI - Detection of apoptosis in hair follicles and acrosyringium of normal human scalp skin by labeling of nick ends of fragmented DNA. PMID- 10367715 TI - Langerhans cells enclosing sunburn cells in acute UV erythema in vivo. PMID- 10367716 TI - Plasma volume changes during and after acute variations of body hydration level in humans. AB - This study examined plasma volume changes (deltaPV) in humans during periods with or without changes in body hydration: exercise-induced dehydration, heat-induced dehydration and glycerol hyperhydration. Repeated measurements of plasma volume were made after two injections of Evans blue. Results were compared to deltaPV calculated from haematocrit (Hct) and blood haemoglobin concentration ([Hb]). Eight well-trained men completed four trials in randomized order: euhydration (control test C), 2.8% dehydration of body mass by passive controlled hyperthermia (D) and by treadmill exercise (60% of their maximal oxygen uptake, VO2max) (E), and hyperhydration (H) by glycerol ingestion. The Hct, [Hb], plasma protein concentrations and plasma osmolality were measured before, during and after the changes in body hydration. Different Hct and [Hb] reference values were obtained to allow for posture-induced variations between and during trials. The deltaPV values calculated after two Evans blue injections were in good agreement with deltaPV calculated from Hct and [Hb]. Compared to the control test, mean plasma volume declined markedly during heat-induced dehydration [-11.4 (SEM 1.7)%] and slightly during exercise-induced dehydration [-4.2 (SEM 0.9)%] (P < 0.001 compared to D), although hyperosmolality was similar in these two trials. Conversely, glycerol hyperhydration induced an increase in plasma volume [+7.5 (SEM 1.0)%]. These results would indicate that, for a given level of dehydration, plasma volume is dramatically decreased during and after heat exposure, while it is better maintained during and after exercise. PMID- 10367717 TI - Non-uniform mechanical activity of quadriceps muscle during fatigue by repeated maximal voluntary contraction in humans. AB - To determine the non-uniform surface mechanical activity of human quadriceps muscle during fatiguing activity, surface mechanomyogram (MMG), or muscle sound, and surface electromyogram (EMG) were recorded from the rectus femoris (RF), vastus lateralis (VL), and vastus medialis (VM) muscles of seven subjects during unilateral isometric knee extension exercise. Time- and frequency-domain analyses of MMG and of EMG fatigued by 50 repeated maximal voluntary contractions (MVC) for 3 s, with 3-s relaxation in between, were compared among the muscles. The mean MVC force fell to 49.5 (SEM 2.0)% at the end of the repeated MVC. Integrated EMG decreased in a similar manner in each muscle head, but a marked non uniformity was found for the decline in integrated MMG (iMMG). The fall in iMMG was most prominent for RF, followed by VM and VL. Moreover, the median frequency of MMG and the relative decrease in that of EMG in RF were significantly greater (P < 0.05) than those recorded for VL and VM. These results would suggest a divergence of mechanical activity within the quadriceps muscle during fatiguing activity by repeated MVC. PMID- 10367718 TI - Effect of exercise training on total daily physical activity in elderly humans. AB - This study examined the effect of 12 weeks of exercise training on daily physical activity in elderly humans. Training consisted of a weekly group session and an individual session with cardio- and weight-stack machines. A group of 15 subjects served as the exercise group [EXER mean age 59 (SD 4) years], and 7 subjects as the controls [CONT mean age 57 (SD 3) years]. Physical activity and physical fitness were measured before the start of training (T), at week 6 and week 12 (T0, T6, T12 respectively) in EXER, and at T0 and T12 in CONT. Physical activity over 14 days was measured using a tri-axial accelerometer and physical fitness was measured during an incremental exercise test. At T12, mean maximal power output had significantly increased in EXER compared to CONT 8 (SD 12) vs -5 (SD 9) W; P < 0.02] and mean submaximal heart rate (at 100 W) had reduced [-10 (SD 7) vs -2 (SD 6) beats x min(-1); P < 0.05]. No differences or changes in physical activity were observed between EXER and CONT. At T6, physical activity on training days was significantly higher than on non-training days (P < 0.001). When the accelerometer output of the training session was subtracted from the accelerometer output on training days, at T12 non-training physical activity was significantly lower than on non-training days (P < 0.004). Accelerometer output of the individual training session at T12 had significantly increased compared to T6 (P < 0.05), whereas, accelerometer output of the group training session had remained unchanged. In conclusion, in elderly subjects an exercise training programme of moderate intensity resulted in an improved physical fitness but had no effect on total daily physical activity. Training activity was compensated for by a decrease in non-training physical activity. PMID- 10367719 TI - Relationships between muscle mitochondrial DNA content, mitochondrial enzyme activity and oxidative capacity in man: alterations with disease. AB - Muscle mitochondrial content is tightly regulated, and requires the expression of both nuclear and mitochondrial genes. In addition, muscle mitochondrial content is a major determinant of aerobic exercise capacity in healthy subjects. The current study was designed to test the hypothesis that in healthy humans, muscle mitochondrial DNA (mtDNA) content is correlated with citrate synthase activity (a representative nuclear-encoded mitochondrial enzyme) and aerobic exercise capacity as defined by whole-body peak oxygen consumption (VO2). Furthermore, it was postulated that these relationships might be altered with disease. Twelve healthy and five paraplegic subjects underwent exercise testing and vastus lateralis muscle biopsy sampling. An additional ten healthy subjects and eight patients with unilateral peripheral arterial disease (PAD) underwent exercise testing and gastrocnemius muscle biopsy sampling. Citrate synthase activity and mtDNA content were positively correlated in the vastus lateralis muscles from the healthy subjects. This relationship was similar in muscle from paraplegic subjects. mtDNA content was positively correlated with peak VO2 in the healthy subjects and in the paraplegic subjects in whom peak VO2 had been elicited by functional electrical stimulation of the muscle. In contrast, the PAD subjects demonstrated higher mtDNA contents than would have been predicted based on their claudication-limited peak VO2. Thus, in healthy humans there are strong relationships between muscle mtDNA content and both muscle citrate synthase activity and peak VO2. These relationships are consistent with coordinant nuclear DNA and mtDNA expression, and with mitochondrial content being a determinant of aerobic exercise capacity. The relationships seen in healthy humans are quantitatively similar in paraplegic patients, but not in patients with PAD, a disease which is associated with a metabolic myopathy. The relationships between mtDNA content, mitochondrial enzyme activities and exercise capacity provide insight into the physiologic and pathophysiologic regulation of muscle mitochondrial expression. PMID- 10367720 TI - Role of xanthine oxidase in delayed lipid peroxidation in rat liver induced by acute exhausting exercise. AB - The aim of this study was to examine whether xanthine oxidase (XOD)-derived hepatic oxidative damage occurs in the main not during but following strenuous exercise. The degree of damage to hepatic tissue catalyzed by XOD was investigated immediately and 3 h after a single bout of exhausting exercise, in allopurinol and saline injected female Wistar rats. Allopurinol treatment resulted in increased hypoxanthine and decreased uric acid contents in the liver compared with the saline treated group, immediately and 3 h after the exercise. Analysis immediately after the exercise showed no changes in hepatic hypoxanthine, uric acid, and thiobarbituric acid-reactive substance (TBARS) contents in the saline treated group, when compared with the resting controls. However, significant increases in uric acid contents in the saline treated livers were observed 3 h after the exercise, relative to the controls. Hepatic TBARS content in the saline treated group were markedly greater than those in both the control and allopurinol treated groups after 3 h of recovery following the exercise. It was concluded that a single bout of exhausting exercise may impose XOD-derived hepatic oxidative damage, primarily during the recovery phase after acute severe exercise. PMID- 10367721 TI - Blood glucose responses in humans mirror lactate responses for individual anaerobic threshold and for lactate minimum in track tests. AB - The equilibrium point between blood lactate production and removal (La-(min)) and the individual anaerobic threshold (IAT) protocols have been used to evaluate exercise. During progressive exercise, blood lactate [La-]b, catecholamine and cortisol concentrations, show exponential increases at upper anaerobic threshold intensities. Since these hormones enhance blood glucose concentrations [Glc]b, this study investigated the [Glc] and [La-]b responses during incremental tests and the possibility of considering the individual glucose threshold (IGT) and glucose minimum (Glc(min)) in addition to IAT and La-(min) in evaluating exercise. A group of 15 male endurance runners ran in four tests on the track 3000 m run (v3km); IAT and IGT - 8 x 800 m runs at velocities between 84% and 102% of v3km; La-(min) and Glc(min) - after lactic acidosis induced by a 500-m sprint, the subjects ran 6 x 800 m at intensities between 87% and 97% of v3km; endurance test (ET) - 30 min at the velocity of IAT. Capillary blood (25 microl) was collected for [La-]b and [Glc]b measurements. The IAT and IGT were determined by [La-]b and [Glc]b kinetics during the second test. The La-(min) and Glc(min) were determined considering the lowest [La-] and [Glc]b during the third test. No differences were observed (P < 0.05) and high correlations were obtained between the velocities at IAT [283 (SD 19) and IGT 281 (SD 21) m. x min(-1); r = 0.096; P < 0.001] and between La-(min) [285 (SD 21)] and Glc(min) [287 (SD 20) m. x min( 1) r = 0.77; P < 0.05]. During ET, the [La-]b reached 5.0 (SD 1.1) and 5.3 (SD 1.0) mmol x l(-1) at 20 and 30 min, respectively (P > 0.05). We concluded that for these subjects it was possible to evaluate the aerobic capacity by IGT and Glc(min) as well as by IAT and La-(min). PMID- 10367722 TI - Autonomic nervous system responses as performance indicators among volleyball players. AB - Complex motor skills require planning and programming before execution. The autonomic nervous system (ANS) is thought to transcribe these central operations at the peripheral level: a motor act is thought to be simultaneously programmed by central and autonomic nervous structures. The aim of this study was to verify that autonomic responses reflect the quality of central motor programming leading to successful or failed performance when subjects are required to perform a complex motor skill. The specificity of the ANS response has already been demonstrated through direct recording from sympathetic fibres. It has also been demonstrated through several mental tasks and closed motor skills such as shooting: ANS responses have been shown to be capable of distinguishing success from failure. The aim of this experiment was to test whether ANS responses are capable of distinguishing two levels of achievement during the performance of a skill involving uncertainty (open skill). The subjects had to intercept a ball on a volleyball court, using the forearm receive and pass technique, in order to pass it on to a moving human target. The results were displayed in terms of accuracy: accurate passes were successful and inaccurate passes missed the target. Six autonomic variables were recorded simultaneously during the task: skin resistance and potential, skin blood flow and temperature, instantaneous heart rate and respiratory frequency. Results showed that autonomic variables were capable of distinguishing success from failure in 22 subjects out of 24. This made it possible to build up autonomic patterns characterising subjects' performances, and to confirm that autonomic functioning may reveal information processing in the central nervous system. Thus, the study of autonomic responses may constitute an inferential model of central nervous system functioning. Such a method could be used as an index for the control of mental preparation. PMID- 10367723 TI - Human isometric force production and electromyogram activity of knee extensor muscles in water and on dry land. AB - This study was designed to determine trial-to-trial and day-to-day reproducibility of isometric force and electromyogram activity (EMG) of the knee extensor muscles in water and on dry land as well as to make comparisons between the two training conditions in muscle activity and force production. A group of 20 healthy subjects (12 women and 8 men) were tested three times over 2 weeks. A measurement session consisted of recordings of maximal and submaximal isometric knee extension force with simultaneous recording of surface EMG from the vastus medialis, vastus lateralis and biceps femoris muscles. To ensure identical measurement conditions the same patient elevator chair was used in both the dry and the wet environment. Intraclass correlation coefficients (ICC) and coefficients of variation (CV) showed high trial-to-trial (ICC = 0.95-0.99, CV = 3.5%-11%) and day-to-day reproducibility (ICC=0.85-0.98, CV=11%-19%) for underwater and dry land measurements of force and EMG in each muscle during maximal contractions. The day-to-day reproducibility for submaximal contractions was similar. The interesting finding was that underwater EMG amplitude decreased significantly in each muscle during maximal (P < 0.01-P < 0.001) and submaximal contractions (P < 0.05-P < 0.001). However, the isometric force measurements showed similar values in both wet and dry conditions. The water had no disturbing effect on the electrodes as shown by slightly lowered interelectrode resistance values, the absence of artefacts and low noise levels of the EMG signals. It was concluded that underwater force and EMG measurements are highly reproducible. The significant decrease of underwater EMG could have electromechanical and/or neurophysiological explanations. PMID- 10367724 TI - Central and peripheral contributions to muscle fatigue in humans during sustained maximal effort. AB - The purpose of this study was to estimate the relative contributions of central and peripheral factors to the development of human muscle fatigue. Nine healthy subjects [five male, four female; age = 30 (2) years, mean (SE)] sustained a maximum voluntary isometric contraction (MVC) of the ankle dorsiflexor muscles for 4 min. Fatigue was quantitated as the fall in MVC. Three measures of central activation and one measure of peripheral activation (compound muscle action potential, CMAP) were made using electromyography (EMG) and electrical stimulation. Measures of intramuscular metabolism were made using magnetic resonance spectroscopy. After exercise, MVC and electrically stimulated tetanic contraction (50 Hz, 500 ms) forces were 22.2 (3.7)% and 37.3 (7.1)% of pre exercise values, respectively. The measures of central activation suggested some central fatigue during exercise: (1) the central activation ratio [MVC/(MVC + superimposed tetanic force)] fell from 0.94 (0.03) to 0.78 (0.09), (2) the MVC/tetanic force ratio fell from 2.3 (0.7) to 1.3 (0.7), and (3) the integral of the EMG (iEMG) signal decreased to 72.6 (9.1)% of the initial value, while the CMAP amplitude was unchanged. Intramuscular pH was associated by regression with the decline in MVC force (and therefore fatigue) and iEMG. The results indicate that central factors, which were not associated with altered peripheral excitability, contributed approximately 20% to the muscle fatigue developed, with the remainder being attributable to intramuscular (i.e., metabolic) factors. The association between pH and iEMG is consistent with proton concentration as a feedback mechanism for central motor drive during maximal effort. PMID- 10367725 TI - Sodium bicarbonate can be used as an ergogenic aid in high-intensity, competitive cycle ergometry of 1 h duration. AB - The aim of this study was to determine whether a dose of 300-mg x kg(-1) body mass of sodium bicarbonate would effect a high-intensity, 1-h maximal cycle ergometer effort. Ten male, well-trained [maximum oxygen consumption 67.3 (3.3) ml x kg(-1) x min(-1), mean (SD)] volunteer cyclists acted as subjects. Each undertook either a control (C), placebo (P), or experimental (E) ride in a random, double-blind fashion on a modified, air-braked cycle ergometer, attached to a personal computer to which the work and power data was downloaded at 10 Hz. Fingertip blood was sampled at 10-min intervals throughout the exercise. Blood was also sampled at 1, 3, 5, and 10 min post-exercise. Blood was analysed for lactate, partial pressure of Carbon dioxide and oxygen, pH and plasma bicarbonate (HCO-) concentration. Randomly chosen pairs of subjects were asked to complete as much work as possible during the 60-min exercise periods in an openly competitive situation. The sodium bicarbonate had the desired effect of increasing blood HCO3 prior to the start of the test. The subjects in E completed 950.9 (81.1) kJ of work, which was significantly more (F(2,27) = 5.28, P < 0.01) than during either the C [835.5 (100.2) kJ] or P [839.0 (88.6) kJ] trials. No differences were seen in peak power or in the power:mass ratio between these three groups. The results of this study suggest that sodium bicarbonate may be used to offset the fatigue process during high-intensity, aerobic cycling lasting 60 min. PMID- 10367726 TI - Biology and pathology of the skin basement membrane zone. PMID- 10367727 TI - Biology and function of hemidesmosomes. AB - Hemidesmosomes are cell-substratum adhesion sites that connect the extracellular matrix to the keratin cytoskeleton. Our knowledge of the function of these structures has greatly increased as a result of studies on patients with aberrant expression of hemidesmosome components and studies using targeted inactivation of mouse genes encoding these components. Insight into the formation of hemidesmosomes, as well as into protein-protein interactions that occur in these junctional complexes, has recently been gained by in vitro cell transfections, blot overlay and yeast two-hybrid assays. In addition, recent results indicate that the alpha6 beta4 integrin is involved in the transduction of signals that are induced by the extracellular matrix and which modulate processes as diverse as cell proliferation, differentiation, apoptosis, migration and tissue morphogenesis. Thus it seems that hemidesmosomes do not merely maintain dermo epidermal adhesion and tissue integrity, but that they are also implicated in intracellular signaling. Here we discuss recently published data on the biology and function of hemidesmosomes. PMID- 10367728 TI - Laminins of the dermo-epidermal junction. AB - Laminins are the most abundant structural non-collagenous glycoproteins ubiquitously present in basement membranes. They are multidomain molecules constituting a family of possibly more than 50 members. Some members such as laminins 5, 6 and 10 are specific of the basal lamina present under stratified epithelia. Although only few intact laminin isoforms have been purified from cultivated cells or tissues, genetic engineering has opened the way for a rapid development of laminin structural biology. Moreover, the phenotypes resulting from gene targeting in mouse or from laminin defects in acquired or inherited human diseases highlight the pivotal role of laminins in morphogenesis, development, and physiology. Indeed, the laminins display a remarkable repertoire of functions, most importantly as structural elements forming a network throughout the basement membrane to which other collagenous or non-collagenous glycoproteins and proteoglycans attach. Furthermore, they are signaling molecules providing adjacent cells with diverse information by interacting with cell surface components. PMID- 10367729 TI - Mutation analysis and molecular genetics of epidermolysis bullosa. AB - Cutaneous basement membrane zone (BMZ) consists of a number of attachment structures that are critical for stable association of the epidermis to the underlying dermis. These include hemidesmosomes, anchoring filaments and anchoring fibrils which form an interconnecting network extending from the intracellular milieu of basal keratinocytes across the dermal-epidermal basement membrane to the underlying dermis. Aberrations in this network structure, e.g. due to genetic lesions in the corresponding genes, can result in fragility of the skin at the level of the cutaneous BMZ. The prototype of such diseases is epidermolysis bullosa (EB), a heterogeneous group of genodermatoses characterized by fragility and blistering of the skin, often associated with extracutaneous manifestations, and inherited either in an autosomal dominant or autosomal recessive manner. Based on constellations of the phenotypic manifestations, severity of the disease, and the level of tissue separation within the cutaneous BMZ, EB has been divided into clinically distinct subcategories, including the simplex, hemidesmosomal, junctional and dystrophic variants. Elucidation of BMZ gene/protein systems and development of mutation detection strategies have allowed identification of mutations in 10 different BMZ genes which can explain the clinical heterogeneity of EB. These include mutations in the type VII collagen gene (COL7A1) in the dystrophic (severely scarring) forms of EB; mutations in the laminin 5 genes (LAMA3, LAMB3 and LAMC2) in a lethal (Herlitz) variant of junctional EB; aberrations in the type XVII collagen gene (COL17A1) in non-lethal forms of junctional EB; mutations in the alpha6 and beta4 integrin genes in a distinct hemidesmosomal variant of EB with congenital pyloric atresia; and mutations in the plectin gene (PLEC1) in a form of EB associated with late onset muscular dystrophy. Identification of mutations in these gene/protein systems attests to their critical importance in the overall stability of the cutaneous BMZ. Furthermore, elucidation of mutations in different variants of EB has direct clinical applications in terms of refined classification, improved genetic counseling, and development of DNA-based prenatal testing in families with EB. PMID- 10367730 TI - Biology of anchoring fibrils: lessons from dystrophic epidermolysis bullosa. AB - Anchoring fibrils are adhesive suprastructures that ensure the connection of the epidermal basement membrane with the dermal extracellular matrix. The fibrils represent polymers of collagen VII, the major structural fibril component, but may also contain other proteins. Remarkable progress has been made in the last few years in understanding the functions of skin basement membrane components including the anchoring fibrils. Novel insights into the biology of the anchoring fibrils have been gained from experimental studies on dystrophic epidermolysis bullosa (DEB), a group of inherited blistering disorders caused by mutations in the gene for collagen VII, COL7A1. Mutation analyses of DEB families have disclosed more than 100 COL7A1 gene defects so far, but the unusual complexity of the mutation constellations and their biological consequences are only beginning to emerge. In analogy to heritable disorders of other collagen genes, predictable phenotypes of COL7A1 mutations causing premature termination codons or dominant negative interference have been observed. However, collagen VII seems to represent a remarkable exception among collagens in that many mutations, including heterozygous glycine substitutions and deletions, lead to minimal phenotypes, or to no phenotype at all. In contrast to fibrillar collagens, structural abnormalities of collagen VII molecules in anchoring fibrils appear to be tolerated to a certain extent. However, the mild DEB phenotypes can be severely modulated by a second aberration in individuals compound heterozygous for two different COL7A1 mutations. Therefore, not only definition of mutation(s) but also cell biological, protein chemical and suprastructural studies of the mutated molecules yield novel insight into the molecular pathomechanisms underlying disease. PMID- 10367731 TI - The matrilins: a novel family of oligomeric extracellular matrix proteins. AB - The matrilin family at present has four members that all share a structure made up of von Willebrand factor A domains, epidermal growth factor-like domains and a coiled coil alpha-helical module. The first member of the family, matrilin-1 (previously called cartilage matrix protein or CMP), is expressed mainly in cartilage. Matrilin-3 has a similar tissue distribution, while matrilin-2 and -4 occur in a wide variety of extracellular matrices. Matrilin-1 is associated with cartilage proteoglycans as well as being a component of both collagen-dependent and collagen-independent fibrils and on the basis of the related structures other matrilins may play similar roles. The matrilin genes are strictly and differently regulated and their expression may serve as markers for cellular differentiation. PMID- 10367732 TI - Metabolic processing of newly synthesized link protein in bovine articular cartilage explant cultures. AB - In explant cultures of articular cartilage from cattle of different ages radiolabeled leucine was shown to be incorporated into link proteins 1, 2 and 3. The newly synthesized link proteins were incorporated into and lost from the cartilage extracellular matrix with time. The levels of radiolabeled link proteins 1 and 2 remaining in the matrix declined over the culture period, but there was an initial increase in the amount of radiolabeled link protein 3, before its level declined. The turnover time of the radiolabeled link proteins 1 and 2 were similar, indicating that neither link protein was preferentially processed to generate link protein 3, nor lost from the extracellular matrix. The majority of the radiolabeled link protein lost from the cartilage matrix could not be recovered from the culture medium, suggesting that turnover of the radiolabeled aggrecan complexes involves the newly synthesized link protein being internalized by the chondrocytes. Inclusion of cytotoxic proteinase inhibitors to the culture medium resulted in a marked decrease in the rate of loss of link protein from the cartilage, suggesting that the catabolism of link protein is cell-mediated and dependent on metabolically active cells. PMID- 10367733 TI - Neurite outgrowth promotion by the alternatively spliced region of tenascin-C is influenced by cell-type specific binding. AB - We have investigated the impact of cellular environment on the neurite outgrowth promoting properties of the alternatively spliced fibronectin type-III region (fnA-D) of tenascin-C. FnA-D promoted neurite outgrowth in vitro when bound to the surface of BHK cells or cerebral cortical astrocytes, but the absolute increase was greater on astrocytes. In addition, different neurite outgrowth promoting sites were revealed within fnA-D bound to the two cellular substrates. FnA-D also promoted neurite outgrowth as a soluble ligand; however, the actions of soluble fnA-D were not affected by cell type. Therefore, we hypothesized that different mechanisms of cellular binding can alter the growth promoting actions of bound fnA-D. We found that fnA-D utilizes two distinct sequences to bind to the BHK cell surface as opposed to the BHK extracellular matrix. In contrast, only one of these sequences is utilized to bind to the astrocyte matrix as opposed to the astrocyte surface. Furthermore, Scatchard analysis indicated two types of receptors for fnA-D on BHK cells and only one type on astrocytes. These results suggest that active sites for neurite outgrowth within fnA-D are differentially revealed depending on cell-specific fnA-D binding sites. Therefore, the function of tenascin-C and its various domains must be considered in terms of cellular context. PMID- 10367734 TI - Cell-type specific and thyroid hormone-dependent expression of genes of alpha1(I) and alpha2(I) collagen in intestine during amphibian metamorphosis. AB - Both the epithelium and the mesenchyme of the larval small intestine of anurans undergoes metamorphic conversion into the adult counterparts. The conversion of the mesenchyme has been poorly understood especially at the molecular level, whereas the changes of the epithelium have been extensively studied. The present study investigated the metamorphic changes of the mesenchyme of tadpoles of bullfrog, Rana catesbeiana, focusing on the expression of genes of type I collagen. By using the cDNA clones coding for a 1(I) and a 2(I) collagen as probes, expression of each collagen gene was examined. These genes were drastically up-regulated at the climax period of spontaneous metamorphosis, which was precociously mimicked by treating tadpoles with thyroid hormone. The increased expression of these genes at the climax stage was well correlated with the conversion of the thin larval mesenchyme to more thick and dense adult connective tissues of the intestine. In situ hybridization identified the fibroblasts that were actively expressing the collagen genes and, therefore, were thought to be responsible for the remodeling. These results strongly suggest that the expression of type I collagen genes is regulated during the intestinal remodeling in a cell-type specific and thyroid hormone-dependent manner. PMID- 10367735 TI - Developing needs led child and adolescent mental health services: issues and prospects. AB - For many years mental health services for children have been developed incrementally with little attention to the needs of the local population. However, over the past decade there have been attempts to develop more rational ways of planning child mental health services. This paper describes the information required to develop a needs-led child mental health service and, within that context, discusses how priorities should be set. It will be suggested that although the assessment of needs for child and adolescent mental health services is still very haphazard, there is now a clear trend for the evaluation of clinical practice to become more systematic. At an individual level we know quite a lot about the efficacy of treatment and the measurement of outcomes. At the service level, several models of good practice are being specified and evaluated. PMID- 10367736 TI - The effect of adverse life events on glycaemic control in children with insulin dependent diabetes mellitus. AB - Forty-five children aged 6 to 14 years with insulin-dependent diabetes mellitus were recruited from a paediatric diabetic clinic. Glycaemic control, child emotional and behavioural problems, and maternal mental state were assessed at recruitment and after 12 months. Life events were measured at 12 months using a standardised semi-structured interview schedule. A between-groups comparison design was used to explore the effect of life events, child emotional and behavioural problems, and maternal mental state on glycaemic control. The children had relatively high rates of emotional and behavioural problems. Overall, the children had a similar number of life events to healthy children in the general population, but proportionally, experienced fewer desirable events. Children who had experienced at least one event in a family setting, or a disappointing event, were more likely to have high glycated haemoglobin levels afterwards than were children who had not experienced such events. Linear regression models showed that only disappointments, family events and glycated haemoglobin at the start of the study made important independent contributions to glycated haemoglobin at 12 months. Child age, emotional and behavioural problems, and maternal mental state, had no effect on glycaemic control. PMID- 10367737 TI - Competencies and problems of Irish children and adolescents. AB - This study set out to investigate the behavioural and emotional problems and competencies of Irish children and adolescents using Achenbach's Child Behaviour Checklist (CBCL) and Youth Self Report (YSR). The Child Behaviour Checklist was completed by parents of 481 Irish school children aged 7-9 years and 13-15 years, and the Youth Self Report was completed by 240 adolescents. The schools were selected to represent a wide social and cultural spread. Irish young people of all ages scored significantly lower than their American counterparts on measures of competence, whether rated by themselves or their parents. The parents of the 7 9 year olds rated their children as having significantly lower total problem scores than their American counterparts, but for 13-15 year olds there were no differences in total problem scores between the Irish and American samples, whether rated by parents or the adolescents themselves. Total problem scores and externalizing scores increased with age on the CBCL and the YSR, a pattern in which Irish young people differed from those in most other cultures. Despite differences in sampling and methodology, the Irish results are similar in many respects to those seen in a number of other European studies. PMID- 10367738 TI - Social competence and emotional/behaviour problems in 6-16 year-old Swedish school children. AB - Social competence and emotional/behavioural problems as reported by parents on a Swedish version of the Child Behaviour Checklist (CBCL) were examined in 1308 Swedish school-aged children/adolescents recruited from a stratified, random sample of schools in urban, semirural, and rural areas in Uppsala County, Sweden, and from Stockholm, the capital city of Sweden. The overall response rate was 80.6%. Few gender differences were found, but adolescents received higher problem scores and higher social competence scores than the younger children. Children from the middle SES groups were regarded as having higher social competence levels, and children from the lower SES groups had higher emotional/behaviour problem scores. Children from the larger cities consistently obtained higher problem scores. Those who had received help during the previous year because of psychological problems (2%) had much higher problems scores than those who had not received help. The levels of emotional/behavioural problems in children and adolescents in the present sample seem to be comparable to those reported in similar Scandinavian studies where the CBCL has been used. However, they were considerably lower than those commonly reported in epidemiological studies of children/adolescents from other countries and cultures. PMID- 10367739 TI - Children with psychiatric disorders who are frequent attenders to primary care. AB - Child psychiatric disorder has been found to be linked to enhanced primary care attendance. We studied the somatic and psychological associations of psychiatric disorder amongst frequent (four or more consultations a year) primary care attending school children. We compared 32 children aged 7-12 years with a psychiatric disorder with 77 non-disordered (also frequently attending) children. Psychiatric disorder was not associated with type of presenting complaint at the surgery nor with chronic physical illness. However disordered children were more likely to be described by their mothers as handicapped by existing physical problems, in poor health, with low energy levels and likely to experience physical symptoms under stress. Problems in social relationships and educational difficulties were reported in more disordered children; more of them came from broken homes and had mothers who reported other psycho-social and health stresses and showed characteristic health beliefs. The findings indicate that knowledge about the child's general physical well-being and relationships and about maternal mental health may assist in the primary care identification and management of psychiatric disorders of frequently attending schoolchildren. PMID- 10367740 TI - The script model in relation to autism. AB - The primary purpose of this study was to investigate autistic children's scripts for social routines. Scripts specify familiar events in terms of who does what, when, to whom, and why. Scripts are verbalizations of mental event representations, containing and organizing generalized knowledge of how the world works. Scripts are presumed to be of vital importance for the development of shared meaning, communication, and social behaviour. In this study, children with autism were asked to explain well-known social routines, such as how you shop in a supermarket, make a cake or celebrate a birthday. The scripts of the 12 children with non-retarded autism were compared to scripts of matched normal control children. Despite the fact that all of the participating children with autism had an IQ above 90 and a mental age between 8 and 14, a significant difference in autistic and normal control children's ability to generate scripts for familiar social routines was found. The results are discussed in relation to the same children's ability to pass theory-of-mind tests and their verbal intelligence. PMID- 10367741 TI - Autistic symptoms in children with attention deficit-hyperactivity disorder. AB - Children with the syndrome of disorders of attention, motor control and perception (DAMP) invariably fill diagnostic criteria for attention deficit hyperactivity disorder (ADHD) and commonly have symptoms of autistic spectrum disorders. This study estimates the rate of autistic symptoms in a sample of children with ADHD by using the parent-rated Autism Criteria Checklist. A high proportion of parents (between 65-80%) reported significant difficulties in social interaction (particularly in empathy and peer relationships), and communication (particularly in imaginative ability, nonverbal communication and maintaining conversation). The nature and relationship between ADHD and pervasive developmental disorders is considered, as well as implications for assessment, diagnosis and treatment. PMID- 10367742 TI - Child and adolescent psychiatry in The Netherlands: patterns of practice. PMID- 10367743 TI - Bcl-2 inhibits early apoptotic events and reveals post-mitotic multinucleation without affecting cell cycle arrest in human epithelial tumor cells exposed to etoposide. AB - Defective apoptotic mechanisms are considered to play a role in both the development of malignancy and resistance to chemotherapeutic drugs. The Bcl-2 family of proteins regulate the cellular commitment to survive or die when challenged with various apoptotic stimuli. PURPOSE: The purpose of this study was to identify the point at which Bcl-2 interrupts the apoptotic cascade initiated following exposure of human tumor cells to etoposide. METHODS: A stable Bcl-2 expressing HeLa-transfected clonal cell line, along with its control-vector transfected counterpart, were utilized in this study. Following etoposide exposure, cells were examined for cell cycle arrest, formation of hyperdiploid cells, apoptotic DNA degradation, loss of plasma membrane integrity, levels of expression of members of the Bcl-2 protein family, caspase activation, degradation of poly(ADP-ribose) polymerase and movement of Bax from cytosol to cellular membrane fractions. RESULTS: Caspase activation, poly(ADP-ribose) polymerase degradation and Bax membrane insertion were initiated rapidly following etoposide removal, concomitantly with cell cycle arrest. Whereas Bcl-2 had no effect on etoposide-induced cell arrest, it interrupted all aspects of apoptosis, including activation of caspases, poly(ADP-ribose) polymerase degradation, DNA fragmentation and loss of plasma membrane integrity. Surprisingly, Bcl-2 also blocked Bax membrane insertion. In addition, Bcl-2 also prevented the increase in cellular levels of Bak, Bax and Bcl-xL, along with degradation of actin and Bax. However, inhibition of etoposide-induced apoptosis by Bcl-2 resulted in the accumulation of giant, multinucleated cells that eventually lost the ability to exclude trypan blue without apoptotic morphology or DNA degradation. CONCLUSIONS: These results indicate that biochemical apoptotic processes are initiated concomitant with etoposide-induced cell cycle arrest and are interrupted by Bcl-2 overexpression. However, the aberrant mitotic events induced by etoposide are sufficient to kill these cells even in the absence of apoptosis. PMID- 10367744 TI - Pharmacokinetics and pharmacodynamics of a novel protein kinase inhibitor, UCN 01. AB - PURPOSE: 7-Hydroxystaurosporine (UCN-01) is a potent protein kinase inhibitor and is being developed as a novel anticancer agent. We describe here its pharmacokinetics and pharmacodynamics in experimental animals. METHODS: The pharmacokinetics of UCN-01 were studied following intravenous (i.v.) administration to mice, rats and dogs at doses of 1-9, 0.35-3.5 and 0.5 mg/kg, respectively. We also studied the pharmacodynamics of UCN-01 (9 mg/kg per day) during and after five consecutive i.v. administrations to nude mice bearing xenografted human pancreatic tumor cells (PSN-1). The concentrations of UCN-01 in plasma and tumor were measured by HPLC using a fluorescence detector. RESULTS: UCN-01 in plasma after i.v. administration was eliminated biphasically in mice and rats, and triphasically in dogs. The elimination half-lives in mice, rats and dogs were 3.00-3.98, 4.02-4.46 and 11.6 h, respectively. The total clearance (Cl(total)) values in mice, rats and dogs were high (1.93-2.64, 2.82-3.86 and 0.616 l/h per kg, respectively). The hepatic clearance (Cl(hepatic)) in rats represented 54.0-81.3% of Cl(total). The volumes of distribution at steady-state in mice, rats and dogs were large (7.89-8.42, 13.0-16.9 and 6.09 l/kg, respectively). These pharmacokinetic parameters were dose-independent in mice and rats. UCN-01 produced significant inhibition of tumor growth during five consecutive i.v. administrations in mice bearing the xenografted PSN-1 cells, and the inhibitory effect continued for 3 days after the final administration. UCN-01 concentrations in tumor tissue were much higher than those in the plasma, and the ratio of tumor to plasma concentrations was about 500 at 24 h after five consecutive doses. CONCLUSIONS: The pharmacokinetic studies showed that UCN-01 has a high clearance and large distribution volume in various experimental animals, and its disposition is linear over the range of doses tested. The pharmacodynamic study showed that UCN-01 is distributed at much higher concentrations in tumor than those in plasma and that it significantly inhibits tumor growth. The high distribution of UCN-01 into tumor cells may contribute to the potent inhibition of tumor growth in vivo. PMID- 10367745 TI - Pharmacokinetics and biotransformations of oxaliplatin in comparison with ormaplatin following a single bolus intravenous injection in rats. AB - PURPOSE: Traditionally ultrafilterable Pt has been used to estimate the body exposure to platinum drugs. However, previous studies have shown that ultrafilterable Pt consists of both cytotoxic and inert biotransformation products of platinum drugs. Therefore, it has been proposed that pharmacokinetic parameters of the parent drug and its cytotoxic biotransformation products are more likely to be correlated with the drug toxicity and efficacy than those of ultrafilterable Pt. Oxaliplatin and ormaplatin are likely to form very similar biotransformation products in vivo based on previous studies. However, ormaplatin causes severe and irreversible neurotoxicity while oxaliplatin causes moderate and reversible neurotoxicity. To evaluate the hypothesis that the neurotoxicity is associated with the pharmacokinetics of active biotransformation products, we investigated the biotransformations and pharmacokinetics of oxaliplatin and ormaplatin in rats at equimolar doses. METHODS: 3H-oxaliplatin and 3H-ormaplatin were administered to Wistar male rats through single bolus i.v. injections (20 micromol/kg). Blood was sampled from 3.5 min to 360 min and centrifuged at 2000 g to separate the plasma from red blood cells (RBCs). The RBCs were sonicated and centrifuged at 13000 g to separate the cytosol from the membrane fraction. Both plasma and RBC cytosol were filtered through YMT30 membranes (Mr = 30000 kDa), and the ultrafiltrates were analyzed using a single column HPLC technique to identify and quantitate the biotransformation products. The pharmacokinetics of oxaliplatin, ormaplatin, and their biotransformation products were characterized utilizing the curve stripping and nonlinear least-squares fitting program RSTRIP. RESULTS: The decays of total, plasma, plasma ultrafilterable (PUF), RBC-bound, and plasma protein-bound Pt-dach (only Pt species with an intact dach carrier ligand were quantitated in this study) were described by biphasic curves. No significant kinetic differences between oxaliplatin and ormaplatin were observed for total, plasma, and PUF Pt-dach in the initial alpha decay phase. However, Pt dach bound to plasma proteins fourfold more quickly for ormaplatin than for oxaliplatin, and the AUC for Pt-dach bound to plasma proteins was twofold higher for ormaplatin than for oxaliplatin. The concentration of RBC-bound Pt-dach was highest at the initial time-point of 3.5 min for both drugs, which suggested a very rapid RBC uptake. The binding of Pt-dach to RBCs was slightly greater initially for ormaplatin than for oxaliplatin. However, the RBC-bound Pt-dach decayed more rapidly for ormaplatin (t(1/2alphaRBC) = 5.1 min) than for oxaliplatin (t(1,2alphaRBC) = 15.3 min). Thus the AUC(RBC) was slightly greater for oxaliplatin than for ormaplatin. The AUC was also slightly greater for oxaliplatin than for ormaplatin for the Pt-dach associated with the RBC membrane and RBC cytosolic proteins. However, there was no significant difference between oxaliplatin and ormaplatin for Pt-dach in the RBC cytosolic ultrafiltrate. There was also no significant difference in the AUCpuf between oxaliplatin and ormaplatin. Both oxaliplatin and ormaplatin produced the same types of major plasma biotransformation products including Pt(dach)Cl2, Pt(dach)(Cys)2, Pt(dach)(GSH)2, Pt(dach)(GSH), Pt(dach)(Met), and free dach. The decays of oxaliplatin, ormaplatin, and their biotransformation products were described by biphasic curves. (ABSTRACT TRUNCATED) PMID- 10367746 TI - Comparative neurotoxicity of oxaliplatin, ormaplatin, and their biotransformation products utilizing a rat dorsal root ganglia in vitro explant culture model. AB - PURPOSE: Neurotoxicity is one of the major toxicities of platinum-based anticancer drugs, especially oxaliplatin and ormaplatin. It has been postulated that biotransformation products are likely to be responsible for the toxicity of platinum drugs. In our preceding pharmacokinetic study, both oxaliplatin and ormaplatin were observed to produce the same types of major plasma biotransformation products. However, while the plasma concentration of ormaplatin was much lower than that of oxaliplatin at an equimolar dose, one of their common biotransformation products, Pt(dach)Cl2, was present at 29-fold higher concentrations in the plasma following the i.v. injection of ormaplatin than of oxaliplatin. Because ormaplatin has severe neurotoxicity and Pt(dach)Cl2 is very cytotoxic, we have postulated that Pt(dach)Cl2 is likely to be responsible for the differences in neurotoxicity between ormaplatin and oxaliplatin. In order to test this hypothesis, we compared the neurotoxicity of oxaliplatin, ormaplatin, and their biotransformation products. Since the dorsal root ganglia (DRGs) have been suggested to be the likely targtet for platinum drugs and in vitro DRG explant cultures have been suggested to be a valid model for studying cisplatin associated neurotoxicity, our comparative neurotoxicity study was conducted with DRG explant cultures in vitro. METHODS: Based on the previous studies of cisplatin neurotoxicity, we established our in vitro DRG explant culture utilizing DRGs dissected from E-19 embryonic rats. Rat DRGs were incubated for 30 min with different platinum compounds to mimic in vivo exposure conditions; this was by followed by a 48-h incubation in culture medium at 37 degrees C. At the end of the incubation, the neurites were fixed and stained with toluidine blue, and neurite outgrowth was quantitated by phase-contrast microscopy. The inhibition of neurite outgrowth by platinum compounds was used as an indicator of in vitro neurotoxicity. Since an in vivo study has indicated that the order of neurotoxicity is ormaplatin > cisplatin > oxaliplatin > carboplatin as measured by morphometric changes to rat DRGs, we initially validated our DRG explant culture model by comparing the in vitro neurotoxicity of ormaplatin, cisplatin, oxaliplatin, and carboplatin. After observing the same neurotoxicity rank between this study and a previous in vivo study, we further compared the neurotoxicity of oxaliplatin, ormaplatin, and their biotransformation products including Pt(dach)Cl2, Pt(dach)(H2O)Cl, Pt(dach)(H2O)2, Pt(dach)(Met), and Pt(dach)(GSH) utilizing the DRG explant culture model. RESULTS: Our study indicated that Pt(dach)Cl2 and its hydrolysis products were more potent at inhibiting neurite outgrowth than the parent drugs oxaliplatin and ormaplatin. In contrast, no detectable inhibition of neurite outgrowth was observed for DRGs dosed with Pt(dach)(Met) and Pt(dach)(GSH). CONCLUSION: This study suggests that biotransformation products such as Pt(dach)Cl2 and its hydrolysis products are more neurotoxic than the parent drugs oxaliplatin and ormaplatin. The different neurotoxicity profiles of oxaliplatin and ormaplatin are more likely due to the different plasma concentrations of their common biotransformation product Pt(dach)Cl2 than to differences in their intrinsic neurotoxicity. PMID- 10367747 TI - Phase I study of the cytotoxic agent N-[2-(dimethylamino)ethyl]acridine-4 carboxamide. AB - N-[2-(Dimethylamino)ethyl]acridine-4-carboxamide (DACA) is a new DNA intercalating drug with a dual mode of cytotoxic action that is thought to involve topoisomerases I and II. On the basis of novelty of action and promising preclinical activity against solid tumours in mice, DACA was selected for clinical trial under the auspices of the Cancer Research Campaign, United Kingdom. We report the phase I findings of a 3-h infusion regimen, repeated 3 weekly, of escalating doses through 18-1000 mg/m2 given to 31 patients with solid malignancies. A maximum tolerated dose (MTD) of 750 mg/m2 was identified, with 3 of 6 cycles being abandoned at 1000 mg/m2. Dose-limiting toxicity took the form of infusional arm pain, in some cases associated with facial discomfort, that was of rapid onset and subsided quickly on the cessation of infusion. The mechanism is unclear but is modulated to some extent by the rate of drug delivery, and it was unaffected in this study by concurrent anti-inflammatory or opiate medication. No host or tumour anti-proliferative activity was observed at these doses, and only minimal toxicity of any other kind was evident. Animal data suggest that the MTD achieved with this schedule may be sub-therapeutic in humans. It is therefore important that efforts be continued to explore methods of giving higher doses of DACA. PMID- 10367748 TI - Plasma pharmacokinetics of N-[2-(dimethylamino)ethyl]acridine-4-carboxamide in a phase I trial. AB - DACA [N-[2-(dimethylamino)ethyl]acridine-4-carboxamide] is an acridine derivative with high activity against solid tumours in mice and a dual mode of cytotoxic action involving topoisomerases I and II. The plasma pharmacokinetics of DACA were studied in 28 patients with solid tumours in a phase I trial. A single dose was given every 3 weeks, being escalated from a starting dose of 18 mg/m2 (as the dihydrochloride trihydrate salt) to a maximal dose, limited by severe pain in the infusion arm, of 1000 mg/m2. Drug was given by constant intravenous infusion with a target delivery period of 3 h. Blood samples were taken from the contralateral arm before, during and for up to 72 h after the infusion. DACA was separated from plasma by solid-phase extraction and was analysed by reversed-phase high performance liquid chromatography (C18 column) using fluorescence detection. A two-compartment pharmacokinetic model provided the best fit for the concentration time profiles obtained for most patients showing clearance of 1.00+/-0.36 l h(-1) kg(-1), a volume of distribution of the central compartment of 0.72+/-0.55 l/kg, an initial half-life of 0.28+/-0.19 h and a terminal half-life of 2.04+/-0.94 h. All pharmacokinetic parameters were independent of dose, indicating first-order kinetics. As DACA binds strongly to alpha1-acid glycoprotein, plasma concentrations of this protein were determined and used to estimate free-drug fractions in plasma. Estimated values for the free fraction varied from 0.9% to 3.3% and were lower than those determined by equilibrium dialysis for mice and rats (15% and 16%, respectively). At the maximum tolerated dose (MTD) of 750 mg/m2, the area under the drug concentration-time curve (AUC) was 46.2+/-4.4 microM h, exceeding that obtained in mice treated at the MTD (23.4 microM h). On the other hand, the corresponding free-drug AUC was 0.92+/-0.03 microM h, much lower than the corresponding value (3.5 microM h) determined for mice. These results suggest that free-drug rather than total drug concentrations are more appropriate for interspecies dose comparisons when significant differences exist in the free plasma fraction. PMID- 10367749 TI - Metabolism of N-[2-(dimethylamino)ethyl]acridine-4-carboxamide in cancer patients undergoing a phase I clinical trial. AB - N-[2-(Dimethylamino)ethyl]acridine-4-carboxamide (DACA) is an experimental antitumour agent that has just completed phase I clinical trials in New Zealand and the United Kingdom. Urine (0-72 h) was analysed from 20 patients receiving DACA infused over 3 h (dose range 60-1000 mg/m2, the latter being the highest dose achieved in the trial). Aliquots were analysed for DACA and its metabolites by high-performance liquid chromatography (HPLC). Over 72 h, 44+/-5% (range 20 60%) of the dose was recovered in the urine, with 0.8+/-0.3% (range 0-3.1%) occurring as DACA. The major urinary metabolite was DACA-N-oxide-9(10H)acridone, accounting for 34+/-3% of the dose. Minor metabolites were identified as N monomethyl-DACA-9(10H)acridone (2.0+/-0.5%), DACA-9(10H)acridone (3.3+/-0.5%), N monomethyl-DACA (0.2+/-0.1%) and DACA-N-oxide (0.5+/-0.1%). No ring-hydroxylated metabolite was detected. The urinary excretion of metabolites was greatest over 0 6 h in most patients. The composition of urinary metabolites was also independent of the delivered dose. Plasma was sampled at intervals throughout the infusion and at time points up to 48 h post-administration. The major plasma metabolites observed were DACA-9(10H)acridone and DACA-N-oxide-9(10H)acridone. These results indicate that, based on urinary excreted metabolites, the major biotransformation reactions for DACA in humans involve N-oxidation of the tertiary amine side chain and acridone formation, both of which appear to be detoxication reactions. PMID- 10367750 TI - Cellular sensitization to cisplatin and carboplatin with decreased removal of platinum-DNA adduct by glucose-regulated stress. AB - PURPOSE: Stress conditions, such as glucose starvation and hypoxia, that induce glucose-regulated proteins (GRPs) in cells, are seen in most solid tumors. These conditions have been shown to cause cellular resistance to multiple anticancer drugs, such as etoposide, doxorubicin, and camptothecin. We examined the effect of the GRP-inducing conditions on cellular sensitivity to cisplatin and carboplatin, which are widely used drugs against solid tumors. METHODS: We generated the GRP-inducing culture conditions by exposing cells to 2-deoxyglucose (2DG), calcium ionophore A23187 and tunicamycin, and examined cellular sensitivity to cisplatin and carboplatin under these conditions. We next measured platinum accumulation and DNA-bound platinum in 2DG-stressed cells after cisplatin exposure. RESULTS: The GRP-inducing stress conditions led to cellular sensitization to cisplatin and carboplatin. This sensitization was reversible, as the cellular sensitivity returned to normal levels 12 h after removal of 2DG. Platinum accumulation and DNA-bound platinum that were found immediately after exposure to cisplatin for 1 h were slightly increased in 2DG-stressed cells as compared with nonstressed cells. After a drug-free recovery incubation of 8 h, the DNA-bound platinum in the nonstressed cells was reduced by 33% while the amount in the 2DG-stressed cells was sustained at the initial levels. CONCLUSIONS: These results indicated that the decreased removal of platinum-DNA adducts was associated with increased sensitivity to cisplatin and carboplatin in the stressed cells. The sensitization of cancer cells under the GRP-inducing stress conditions would explain, in part, the clinical potency of platinum drugs against solid tumors. PMID- 10367751 TI - Local disposition kinetics of floxuridine after intratumoral and subcutaneous injection as monitored by [19F]-nuclear magnetic resonance spectroscopy in vivo. AB - PURPOSE: To test the utility of [19F]-nuclear magnetic resonance (NMR) spectroscopy for studying the kinetics of local drug disposition after interstitial application in vivo. METHODS: Floxuridine at 30 micromol (2.5% of the reported i.p. 50% lethal dose, LD50) was injected into rats either intratumorally (Morris hepatoma M3924A) or s.c. [19F]-NMR spectra were obtained at the site of administration for up to 5 h after injection using a 2-cm diameter surface coil at 2.0 T. Signal-time data obtained for floxuridine and the metabolite 5-fluorouracil were analyzed using linear compartment models. RESULTS: The lower limit for the quantitation of drug remaining at the site of administration was 1 micromol for tumors and 0.2 micromol for the s.c. injection site. Local drug disposition was biexponential in four of six tumors where the half-lives of the fast and slow components of disposition ranged from 4 to 26 and from 33 to 289 min, respectively. It was monoexponential in the remaining two tumors (half-lives 49 and 128 min) and in the s.c. injection experiments (n = 4, half-life 6-9 min). 5-Fluorouracil could be quantitated in three of six tumors; the estimated fraction of floxuridine converted intratumorally into 5 fluorouracil was 11-23%. Alpha-fluoro-beta-alanine was detected in the sum spectra of three of the six tumours. CONCLUSIONS: Local drug-disposition kinetics after interstitial application can be monitored noninvasively by in vivo [19F] NMR spectroscopy. Disposition kinetics after local injection is highly variable and has a slow component in this tumor, whereas it is much less variable and relatively fast in subcutaneous tissue. The results suggest that NMR spectroscopy may be useful for in vivo studies of drug release from depot preparations designed for interstitial application. PMID- 10367752 TI - Phase I and pharmacokinetic analysis of high-dose tamoxifen and chemotherapy in normal and tumor-bearing dogs. AB - PURPOSE: To determine whether tamoxifen plasma concentrations capable of blocking P-glycoprotein (Pgp) in vitro can be safely achieved in dogs and whether doxorubicin pharmacokinetic alterations occur when tamoxifen is coadministered. METHODS: Tamoxifen dose escalation studies were conducted in 7 normal dogs and in 19 tumor-bearing dogs receiving full-dose chemotherapy. Plasma tamoxifen and serum doxorubicin disposition were analyzed for putative drug interactions. RESULTS: Steady-state plasma concentrations of tamoxifen and N-desmethyl tamoxifen (NDMT) were 5-10 microM following oral tamoxifen administration at 600 mg/m2 every 12 h for 7 days to normal and tumor-bearing dogs. Mild-moderate gastrointestinal toxicity (diarrhea, anorexia) and reversible neurotoxicity were observed in dogs receiving chemotherapy plus high-dose tamoxifen. Myelosuppression was not affected by combined treatment in tumor-bearing dogs. High-dose tamoxifen decreased the clearance and volume of distribution of full dose doxorubicin. CONCLUSIONS: Concentrations of tamoxifen/ NDMT sufficient to inhibit Pgp may be achieved in dogs receiving full-dose chemotherapy with a moderate but acceptable increase in gastrointestinal toxicity. Tamoxifen affects doxorubicin metabolism in dogs at high doses resulting in increased serum exposure. Pharmacologic manipulation of Pgp expression or function in normal and tumor tissue in dogs may facilitate investigation of novel anticancer treatment strategies in humans. PMID- 10367753 TI - In vitro evaluation of flavopiridol, a novel cell cycle inhibitor, in bladder cancer. AB - PURPOSE: To determine the in vitro effects of flavopiridol on bladder cancer cell lines, immortalized urothelial cell lines, and normal urothelial cells well characterized for defects in p53, pRb, and p16. METHODS: Growth inhibition was assessed via an MTT assay and apoptosis via DAPI nuclear staining. Cell cycle analysis was performed via propidium iodide staining and fluorescent activated cell sorting (FACS). Multidrug-resistant cells were generated by continuous exposure to doxorubicin. RESULTS: Growth inhibition was not correlated with inactivation of p53, pRb, or p16. All cells experienced G2/M arrest within 24 h of flavopiridol exposure. Modest apoptosis was observed but required 72 h of continuous drug exposure to become evident. There was no obvious synergistic or antagonistic toxicity when flavopiridol was combined with radiotherapy or cisplatin dosed at the IC50 despite the observation that radiotherapy and flavopiridol led to more profound G2/M arrest than either agent alone. Doxorubicin-resistant cells, demonstrated to overexpress the MDR1 multi-drug resistance protein were equally as sensitive to flavopiridol as the parental cells. CONCLUSIONS: Flavopiridol is a novel cell cycle inhibitor that may be a useful agent in bladder cancers with tumor suppressor gene alterations and/or multidrug resistance. PMID- 10367754 TI - Omeprazole does not alter plasma methotrexate clearance. PMID- 10367755 TI - An analytical model of lumbar motion segment in flexion. AB - OBJECTIVE: To develop an analytical model of the lumbar motion segment and to determine the following under the application of flexion physiological loads: (1) the force displacement relationships of the lumbar motion segment; (2) the forces in the ligaments, disc, and facet joints; (3) the strains in the ligaments; and (4) the effect of the transection of the ligaments. DESIGN: Computer modeling. SETTING: Spinal Ergonomics and Joint Research Laboratory at The National College of Chiropractic. PROCEDURE: Computer model simulation of external loads and simulation of ligament transection. MEASURES: The following parameters were predicted in flexion by means of a computer model: (1) the load-displacement relationships of the lumbar motion segment; (2) the loads in the ligaments, disc and facet joints; (3) the strains in the ligaments; and (4) the effect of the transection of the ligaments. RESULTS: The load sharing among different ligaments predicted by this model under flexion load suggests that the supraspinous ligament carries the greatest load, followed by the yellow ligament, capsular ligament, intertransverse ligament, and interspinous ligament. The ligament strains indicate that the supraspinous ligament undergoes the maximum increase in length, followed by the interspinous ligament, yellow ligament, capsular ligament, and intertransverse ligament. The transection of ligaments increased the flexibility of the joint, the strains on the rest of the ligaments, the loads on all of the rest of the ligaments, as well as the moment on the disc, but does not significantly affect the compressive load on the disc. CONCLUSIONS: The analytical model predicts results similar to the experimental data on cadaver motion segments reported in the literature under flexion moment loads. PMID- 10367756 TI - Supervision of chiropractors: a summary of results from two surveys involving chiropractic supervisors and graduates in England and Sweden. AB - BACKGROUND: Supervision of newly graduated health care practitioners takes place in many clinical settings, such as in different types of hospital departments, in general practice, and now also within the chiropractic profession. The author conducted an initial study to determine the most important issues regarding the supervision of chiropractic graduates in Sweden. Because it is important to define and discuss the format and contents for that part of the 1-year postgraduate education program for the newly graduated chiropractor that takes place in private chiropractic offices, a new study was conducted involving chiropractic supervisors and new graduates in Sweden and England. OBJECTIVES: To establish what chiropractic supervisors and new graduates believe is important for the graduates to learn during the clinical part of their postgraduate education, to describe the characteristics required (according to supervisors and graduates in the study) to be a good supervisor for a chiropractic graduate, and to investigate whether there are differences in opinions between supervisors and graduates and between the 2 countries. METHODS: Questionnaires were sent out to 11 Swedish chiropractic graduates, 30 English chiropractic graduates, and 30 English chiropractic supervisors who had chiropractic graduates at the time of the survey. RESULTS: Participants agreed that the most important aspect of the clinical activities was to have regular meetings with the graduate, to be professional, and to explain the patient's problems. The most important characteristic to become a good supervisor was that the supervisor is willing to spend time and listen to the graduate throughout the postgraduate training period. The Swedish participants more often considered it important to give the graduates academic articles to read. CONCLUSION: The results of this study suggest that it is the human aspect and the personal relationship between the supervisor and the graduate that are important during the graduate's 1-year postgraduate education at the chiropractic clinic. Chiropractic supervisors and new graduates both believe that one of the most important aspects of supervision is the ongoing dialogue between the supervisor and the graduate. PMID- 10367757 TI - The frequency of positive common spinal clinical examination findings in a sample of premenstrual syndrome sufferers. AB - OBJECTIVE: As part of a randomized clinical trial to determine the efficacy of chiropractic therapy on premenstrual syndrome (PMS), subjects were evaluated for initial underlying spinal dysfunction. SUBJECTS: Fifty-four subjects with diagnosed PMS (using a Moos PMS questionnaire plus daily symptom monitoring) and 30 subjects with no diagnosable PMS were recruited by newspaper advertising and referrals. DESIGN: All subjects underwent a full history and physical and chiropractic examination carried out by 1 of 2 fully qualified and registered chiropractors, each with a minimum of 10 years experience. The results of the assessment for the PMS group were compared with those of the non-PMS group. SETTING: RMIT teaching clinics. DATA ANALYSIS: The data collected were entered into a spread sheet and contingency tables were created. The data were analyzed by use of chi-squared tests, with the statistical significance being set at P < .05. RESULTS: The PMS group had a higher percentage of positive responses for each of 12 measured spinal dysfunction indexes except for range of motion of the low back. The indexes where the increase was statistically significant (P < .05) were cervical, thoracic, and low back tenderness, low back orthopedic testing, low back muscle weakness, and the neck disability index. An average of 5.4 of the 12 indexes were positive for the PMS group compared with 3.0 for the non-PMS group. CONCLUSIONS: A relatively high incidence of spinal dysfunction exists in PMS sufferers compared with a comparable group of non-PMS sufferers. This is suggestive that spinal dysfunction could be a causative factor in PMS and that chiropractic manipulative therapy may offer an alternative therapeutic approach for PMS sufferers. PMID- 10367759 TI - A review of biomechanics of the central nervous system--Part I: spinal canal deformations resulting from changes in posture. AB - OBJECTIVE: To discuss how the spinal cord deforms as a result of changes in posture or biomechanical alterations of the spine. DATA COLLECTION: A hand search of available reference texts and a computer search of literature from the Index Medicus sources were collected, with special emphasis placed on spinal canal changes caused by various postural rotations and translations of the skull, thorax, and pelvis. RESULTS: All spinal postures will deform the spinal canal. Flexion causes a small increase in canal diameter and volume as the vertebral lamina are separated. Extension causes a small decrease in canal diameter and volume as the vertebral lamina are approximated. Lateral bending and axial rotation cause insignificant changes in spinal canal diameter and volume in cases without stenosis. CONCLUSIONS: Rotations of the global postural components, head, thoracic cage, and pelvis cause changes in the diameter of the spinal canal and intervertebral foramen. These changes are generally a reduction of less than 1.5 mm in extension, compared with a small increase in flexion of approximately 1 mm. These small changes do not account for the clinical observation of patients having increased neurologic signs and symptoms in flexion. PMID- 10367758 TI - Radiographic evaluation of weight-bearing orthotics and their effect on flexible pes planus. AB - OBJECTIVE: To determine whether any positive change in the alignment of the bones of the feet occur with the use of custom-made flexible orthotics, cast by weight bearing, in individuals having flexible pes planus. METHODS: Anteroposterior and lateral radiographs were obtained with and without orthotics in place. The anteroposterior and lateral talocalcaneal angles and the lateral pitch of both the left and right foot were assessed. RESULTS: The t test values and P values derived from the radiographic measurements indicated statistically significant improvements in weight-bearing foot alignment. DISCUSSION: Biomechanical faults in the pedal foundation can adversely affect any of the joints and structures of the foot/ankle complex, lower extremities, pelvis, and spine. CONCLUSION: This study supports the use of a custom-made flexible orthotic for the improvement of pedal structural alignment. PMID- 10367760 TI - Management of acute lumbar disk herniation initially presenting as mechanical low back pain. AB - OBJECTIVE: To describe the clinical management with spinal manipulation of a male patient with risk factors for lumbar disk herniation initially suffering from what appeared to be mechanical low back pain that evolved into radiculopathy; also to review issues pertinent to chiropractic/manipulative management of disk herniation. CLINICAL FEATURES: The patient initially suffered from unilateral low back pain and nonradicular/nonlancinating referral to the ipsilateral lower extremity. INTERVENTION AND OUTCOME: Disk herniation-in-evolution was included in the differential diagnosis, which was discussed with the patient, who then gave verbal informed consent for manipulative management. A day or so after the initial manipulation the presentation evolved to include S1 radiculopathy. Computed tomography, just after onset of radiculopathy, confirmed the clinical diagnosis of lumbosacral disk herniation. The patient continued with manipulative management and repeat computed tomography examination after clinical resolution about 2 months later revealed reduction in size of the apparently clinically significant herniation. CONCLUSION: Risk factors for the development of disk herniation should be considered when assessing patients suffering from what appears to be mechanical low back pain. The role played by manipulation in the development of disk herniation in this case was believed to be circumstantial rather than causal. Manipulation was used in the treatment of this patient over a period of approximately 2 months; after this time, clinical and partial computed tomography imaging resolution was evident. Ongoing clinical (neurologic) evaluation of patients with manifest or suspected disk herniation is an important aspect of management. Good-quality trials of manipulation for patients with disk herniation are imperative for the chiropractic profession. PMID- 10367761 TI - Posterior fossa ischemia and bilateral vertebral artery hypoplasia. AB - OBJECTIVE: To discuss cerebellar infarct in a patient with bilateral hypoplasia of the vertebral arteries. CLINICAL FEATURES: A 67-year-old woman suffered neck pain and headaches immediately after a minor motor vehicle accident. Fracture and dislocation were radiographically ruled out. Within a few days, the patient began to experience symptoms of vertigo and dizziness. Computed tomography scanning of the brain revealed a cerebellar infarct, and an angiogram of the vertebral arteries demonstrated bilateral hypoplasia. INTERVENTION AND OUTCOME: The cerebellar infarct created mild, stable symptoms, and the patient was monitored closely in a hospital setting until the risk of possible complications was negligible. After an uncomplicated and full recovery, the patient was given recommendations regarding critical neck positions to decrease the potential for further ischemic events. CONCLUSION: Cerebellar infarcts are rare and may be associated with rare vascular anomalies. PMID- 10367762 TI - Chiropractic: more good than harm or vice versa? PMID- 10367763 TI - The routine use of radiographic spinal displacement analysis: a dissent. PMID- 10367764 TI - Motion palpation: it's time to accept the evidence. PMID- 10367765 TI - Motion palpation: it's time to accept the evidence. PMID- 10367766 TI - Antimicrobial peptides as mediators of epithelial host defense. AB - Mammalian epithelial surfaces are remarkable for their ability to provide critical physiologic functions in the face of frequent microbial challenges. The fact that these mucosal surfaces remain infection-free in the normal host suggests that highly effective mechanisms of host defense have evolved to protect these environmentally exposed tissues. Throughout the animal and plant kingdoms, endogenous genetically encoded antimicrobial peptides have been shown to be key elements in the response to epithelial compromise and microbial invasion. In mammals, a variety of such peptides have been identified, including the well characterized defensins and cathelicidins. A major source of these host defense molecules is circulating phagocytic leukocytes. However, more recently, it has been shown that resident epithelial cells of the skin and respiratory, alimentary, and genitourinary tracts also synthesize and release antimicrobial peptides. Both in vitro and in vivo data support the hypothesis that these molecules are important contributors to intrinsic mucosal immunity. Alterations in their level of expression or biologic activity can predispose the organism to microbial infection. The regulatory and developmental aspects of antimicrobial peptide synthesis are discussed from a perspective that emphasizes the possible relevance to pediatric medicine. PMID- 10367767 TI - GB virus C (GBV-C/HGV) and E2 antibodies in children preliver and postliver transplant. AB - The association of GB virus type C (GBV-C) virus and clinical disease is uncertain. The role of GBV-C and (Envelope) E2 antibody in children with liver transplants has not been determined. This study's aim is to examine the prevalence of GBV-C in children with liver transplants, to assess the relationship of GBV-C to posttransplant hepatitis, and to determine the role of E2 antibodies. Sera from 34 children, preliver and postliver transplant, between 1989-1996 were tested for GBV-C (Ribonucleic acid) RNA by the automated Abbott LCx PCR assay. Anti-E2 antibodies were detected by an Abbott immunoassay. Recent posttransplant liver biopsies were examined for hepatitis. The results of the study determined that pretransplant, four children (12%) were GBV-C RNA positive. Posttransplant, 14 (42%) children were GBV-C RNA positive. The GBV-C RNA positive conversion rate was 33% (CI 17.2-55.7%). Patients received blood products from a mean of 68 +/- 34 donors, which correlated with GBV-C acquisition. There was no difference in the incidence (32%versus 36%; p = 0.726) or severity (grade 2.00 versus 0.68; p = 0.126) of posttransplant hepatitis in the liver biopsies of GBV C RNA negative and/or positive children, respectively. Pretransplant, nine of 32 children were anti-E2 positive. Posttransplant, eight of 32 children were anti-E2 positive, including five children who were anti-E2 positive pretransplant. Of nine children who were anti-E2 positive and GBV-C RNA negative pretransplant, three became GBV-C RNA positive posttransplant. The results of this study conclude that the prevalence of GBV-C infection in children postliver transplantation is high and that blood product transfusions correlate with GBV-C acquisition. Also, no correlation was found between GBV-C RNA and the incidence or severity of posttransplant hepatitis. Finally, E2 antibody presence before transplantation failed to provide complete protection from GBV-C acquisition. PMID- 10367768 TI - Investigation of seroprevalence of respiratory virus infections in an infant population with a multiantigen fluorescence immunoassay using heel-prick blood samples collected on filter paper. AB - Respiratory viruses are an extremely common cause of childhood morbidity. However, the current seroprevalence of viruses in infant populations is difficult to establish because invasive venipuncture may be technically and ethically unacceptable. This prospective study aimed to establish the seroprevalence of respiratory viruses in an infant population by use of a novel multiantigen fluorescence immunoassay against common respiratory viruses, using heel-prick blood samples collected on filter paper. Mothers and babies were recruited in the immediate peripartum period in the Royal Maternity Hospital, Belfast. Cord blood samples at birth and heel-prick filter paper blood samples at 7 mo were collected for measurement of virus-specific IgG to respiratory syncytial virus, influenza A virus, adenovirus, and parainfluenza virus type 1, type 2, and type 3 by indirect immunofluorescence using a multiviral assay developed for this purpose. Of 386 mothers approached, 325 (84%) permitted follow-up at 7 mo, and of these, 256 (79%) agreed to the heel prick. From 234 paired samples, 125 infections were documented. Adenovirus infections were commonest, 53 (22.6%), followed by respiratory syncytial virus, 32 (13.7%); influenza A virus, 22 (9.4%); parainfluenza virus type 3, 14 (6%); parainfluenza virus type 1, 2 (0.85%); and parainfluenza virus type 2, 2 (0.85%). These results demonstrate the seroprevalence of a range of respiratory viruses in an infant population, using a novel multiviral immunoassay. The filter paper collection of blood samples and multiantigen assay format has implications for easy, widespread viral serodiagnosis in both seroepidemiology studies and in the diagnosis of pediatric viral illnesses. Filter paper permits recovery of respiratory virus-specific IgG and can be used as a simple and acceptable epidemiologic and diagnostic tool. PMID- 10367769 TI - IL-6 cerebrospinal fluid levels are related to laryngeal IgA and epithelial HLA DR response in sudden infant death syndrome. AB - The objective was to investigate whether there is any correlation between signs of central and peripheral immune stimulation in victims of sudden infant death syndrome (SIDS), the former expressed by IL-6 in cerebrospinal fluid (CSF), the latter by IgA, IgG, and IgM immunocytes, T lymphocytes, and HLA-DR expression in laryngeal mucosa. Seventeen SIDS cases with low CSF IL-6 levels (< or =5 pg/mL) and 20 cases with high CSF IL-6 levels (> or =30 pg/mL) were subjected to immunohistochemical quantitation of IgA, IgG, and IgM immunocytes; semiquantitative scoring of T lymphocytes in the mucosa of epiglottis and larynx, and semiquantitative evaluation of HLA-DR expression. SIDS cases with IL-6 levels > or =30 pg/mL had a significantly higher number of IgA immunocytes in laryngeal mucosa (p = 0.007) and in epiglottis (p = 0.03) than cases with IL-6 levels < or =5 pg/mL. Furthermore, laryngeal HLA-DR expression was significantly more extensive in SIDS cases with IL-6 levels > or =30 pg/mL than in those with levels < or =5 pg/mL (p = 0.05). No differences were found for IgG and IgM immunocytes or for T cells. In addition, babies found prone more often had symptoms of slight infection before death and had a higher number of IgA immunocytes in the larynx (p = 0.02) than babies sleeping on their side or back. Because IL-6 levels > or =30 pg/mL correspond to the levels found in infants who die from infectious diseases such as meningitis/septicemia and pneumonia, the findings favor the hypothesis that many SIDS cases may be caused by an "overreaction" of the immune system to an otherwise harmless infection. PMID- 10367770 TI - System A amino acid transporter activity in human placental microvillous membrane vesicles in relation to various anthropometric measurements in appropriate and small for gestational age babies. AB - Fetal growth and development is dependent on the transfer of amino acids from maternal to fetal blood across the microvillous plasma membrane (MVM) and basal plasma membrane of placental syncytiotrophoblast. The aim of this study was to determine the relationship of system A amino acid transporter (SysA) activity in MVM to a variety of measurements of size at birth in a group of term small for gestational age (SGA) babies and in a group of appropriate for gestational age (AGA) babies. Mean SysA activities (nmol/mg vesicle protein/30 s +/- SEM) were: SGA, 0.027 +/- 0.004 (n = 25) and AGA, 0.045 +/- 0.005 (n = 24); p = 0.006. Spearman rank correlations were calculated for SGA (n = 19-25) and AGA (n = 21 24) groups for SysA activity against the following anthropometric measurements: abdominal circumference, birth weight, length, midarm circumference (MAC), head circumference, midarm circumference:head circumference ratio, placental weight (PW), placental ratio (placental weight:birth weight), birth weight:length ratio, Ponderal index (birth weight/length3) and triceps and subscapular skin-fold thicknesses (tsft and ssft). In SGA babies, SysA activity was positively correlated (p < 0.05) with subscapular skin-fold thicknesses (r = 0.48), triceps skin-fold thicknesses (r = 0.42), PW (r = 0.42), and placental ratio (r = 0.46). In AGA babies, the only significant correlation was an inverse one with placental ratio (r = -0.50). These data suggest there are differences in the relationship between placental SysA activity and fetal proportion in term AGA compared with SGA babies. PMID- 10367771 TI - Does increasing dietary linolenic acid content increase the docosahexaenoic acid content of phospholipids in neuronal cells of neonatal rats? AB - The objective of this study was to investigate if increasing maternal dietary linolenic acid (18:3n-3) content, by decreasing the 18:2n-6 to 18:3n-3 ratio, could increase the docosahexaenoic acid (22:6n-3) content in phospholipids of neuronal cells of rat pups at 2 weeks of age. Sprague-Dawley dams at parturition were fed semipurified diets containing decreasing ratios of 18:2n-6 to 18:3n-3 from 21.6:1 to 1:1. During the first 2 weeks of life, the rat pups received only their dam's milk. The fatty acid composition of the pups stomach contents (dam's milk) and the phospholipids from neuronal cells were identified and quantitated by gas-liquid chromatography. The stomach 22:6n-3 content analyzed from the rat pups at 2 weeks of age was altered by the maternal diet. Fatty acid analysis of phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylserine (PS) in neuronal cells of the rat pups showed no significant increase in 22:6n-3 content with increasing 18:3n-3 in the maternal diet (p > 0.05). In contrast, the content of 22:6n-3 in phosphatidylinositol (PI) was significantly increased by change in dietary 18:3n-3 intake from a dietary 18:2n-6 to 18:3n-3 ratio of 7.8:1 to 4.4:1. It is concluded that increasing maternal dietary 18:3n-3 by decreasing the 18:2n-6 to 18:3n-3 ratio does not significantly increase the 22:6n-3 content in PC, PE, and PS in neuronal cells of rat pups at 2 weeks of age. PMID- 10367772 TI - Kynurenic acid in brain and cerebrospinal fluid of fetal, newborn, and adult sheep and effects of placental embolization. AB - Concentrations of the endogenous glutamate receptor antagonist kynurenic acid (KA) were measured in various brain regions and in cisternal cerebrospinal fluid of fetal, newborn, and adult sheep. KA concentrations were significantly higher in the fetal brain and cerebrospinal fluid at 90 and 140 d gestation compared with postnatal ages. In fetuses of 132-139 d gestation, KA concentrations in cerebrospinal fluid collected by drainage from an indwelling cisternal catheter increased significantly after infusion of the organic acid transport inhibitor probenecid (100 or 200 mg/kg, i.v.) indicating active transport of KA out of the fetal brain. In fetuses in which the umbilical circulation had been chronically restricted from 120 to 140 d gestation by partial embolization of the placenta, plasma concentrations of the KA precursor kynurenine were significantly lower than in control fetuses, and KA concentrations in the hypothalamus and hippocampus were significantly reduced; other brain regions were not affected. These results indicate that the production of KA is higher in the fetal brain compared with the newborn and adult brain. Because KA diminishes the risk of excitotoxic neuronal damage under hypoxic-ischemic conditions, the high levels of KA in the brain before birth may have a neuroprotective function. The decrease of KA concentrations in the hypothalamus and hippocampus after umbilical embolization suggests that, after chronic hypoxia in utero, these regions of the brain may become more vulnerable to subsequent episodes of acute hypoxia or ischemia encountered in late gestation or during parturition. PMID- 10367773 TI - Effects of hypoxia-ischemia and inhibition of nitric oxide synthase on cerebral energy metabolism in newborn piglets. AB - The present study was designed to examine the effects of inhibition of nitric oxide synthase on cerebral energy metabolism after hypoxia-ischemia in newborn piglets. Ten 1- to 3-d-old piglets received N(omega)-nitro-L-arginine (NNLA), an inhibitor of nitric oxide synthase (NNLA-hypoxia, n = 5), or normal saline (hypoxia, n = 5) 1 h before cerebral hypoxia-ischemia. After the infusion, hypoxia-ischemia was induced by bilateral occlusion of the carotid arteries and decreasing FiO2 to 0.07 and maintained for 60 min. Thereafter, animals were resuscitated and ventilated for another 3 h. Using 1H- and 31P-magnetic resonance spectroscopy, cerebral energy metabolism was measured in vivo at 15-min intervals throughout the experiment. Phosphocreatine to inorganic phosphate ratios decreased from 2.74 +/- 0.14 to 0.74 +/- 0.36 (hypoxia group) and 2.32 +/- 0.17 to 0.18 +/- 0.10 (NNLA-hypoxia group) during hypoxia-ischemia. Thereafter, phosphocreatine to inorganic phosphate ratios returned rapidly to baseline values in the hypoxia group, but remained below baseline values in the NNLA-hypoxia group. Intracellular pH decreased during hypoxia-ischemia and returned to baseline values on reperfusion in both groups. Intracellular pH values were lower in the NNLA-hypoxia group (p < 0.001, ANOVA). Lactate was not present during the baseline period. After hypoxia-ischemia, lactate to N-acetylaspartate ratios increased to 1.34 +/- 0.28 (hypoxia group) and 2.22 +/- 0.46 (NNLA-hypoxia group). Lactate had disappeared after 3 h of reperfusion in the hypoxia group, whereas lactate to N-acetylaspartate ratios were 1.37 +/- 1.37 in the NNLA hypoxia group. ANOVA demonstrated a significant effect of NNLA on lactate to N acetylaspartate ratios (p < 0.001). Inhibition of nitric oxide synthase by NNLA tended to compromise cerebral energy status during and after cerebral hypoxia ischemia in newborn piglets. PMID- 10367774 TI - Cerebrovascular reactivity remains intact after cortical depolarization in newborn piglets. AB - Cerebrovascular reactivity is severely affected by ischemia, and changes in vascular responses have been reported after cortical spreading depression and head trauma as well. Cortical depolarization (CD) occurs during ischemia, cortical spreading depression, and head trauma, but its effects on cerebrovascular reactivity are unclear. We tested the hypothesis that CD induced by KCl diminishes the vascular responsiveness to various vasodilatory stimuli in piglets. Responses of pial arterioles were determined by changes in vascular diameter by use of a closed cranial window and intravital microscopy. Baseline arteriolar diameters were 105 +/- 3 microm (mean +/- SEM, n = 27). CD was elicited by topical administration of 1 mol/L KCl for 3 min. Vascular responses were measured before and 1 h after CD. KCl elicited CD and constricted arterioles by 54 +/- 4% (n = 27). N-methyl-D-aspartate induced dose-dependent vasodilation that was unaffected by CD; the percent changes were 9 +/- 1 versus 8 +/- 1 (before and after CD) at 10(-5) mol/L, 19 +/- 2 versus 18 +/- 3 at 5 x 10(-5) mol/L, and 29 +/- 2 versus 26 +/- 3 at 10(-4) mol/L (n = 9). Hypercapnic vasodilation was not diminished by CD; the percent changes were 15 +/- 2 versus 16 +/- 4 at 5%, and 27 +/- 5 versus 27 +/- 6 at 10% inspired CO2 (n = 8). Aprikalim and forskolin caused dilation that was also resistant to prior CD; the percent change values were 21 +/- 4 versus 18 +/- 3 and 16 +/- 2 versus 16 +/- 4 at 10(-6) mol/L, 36 +/- 5 versus 34 +/- 5 and 34 +/- 7 versus 37 +/- 7 at 10(-5) mol/L (n = 8), respectively. Finally, calcitonin gene-related peptide-induced vasodilation was unaffected by CD; percent changes were 15 +/- 3 versus 16 +/- 2 at 10(-7) mol/L and 26 +/- 4 versus 22 +/- 3 at 10(-6) mol/L (n = 8). The intact vascular responses after CD suggest that this component is not responsible for decreased cerebrovascular reactivity after ischemia, head trauma, or cortical spreading depression. PMID- 10367776 TI - Pacing-induced ventricular remodeling in the chick embryonic heart. AB - Chronic ectopic pacing in the adult heart induces myocardial hypotrophy close to the pacing site. We have recently described a similar localized decrease of compact myocardium thickness in the chick embryonic heart after 48 h of intermittent apical ventricular pacing. Here we analyze the cellular mechanisms underlying the response of the embryonic heart to pacing. Because the developing heart had been found to adjust its morphology according to functional demands by undergoing cellular hyperplasia or hypoplasia, we hypothesized that the stimulation should result in hypoplasia of the apical ventricular compartment. Morphologic analysis of hearts submitted to 18 h of effective pacing during 48 h showed a mild to moderate ventricular dilatation, a 28% decrease in the apical compact layer thickness with no changes in other ventricular locations, and atrial wall thickening. These modifications were caused by changes in the number of cell layers, whereas cell size was similar between paced and control hearts. Analysis of proliferative activity after 24 h of pacing showed a decrease of 32% in the rate of cell proliferation limited to the apical compact layer exposed to stimulation. No ultrastructural injury or increased cell death was found. These changes were accompanied by down-regulation of the myocardial growth factor fibroblast growth factor-2 but no differences were found in the expression of platelet-derived growth factor. Thus, chronic intermittent ventricular pacing induces myocardial remodeling in the chick embryonic heart, on the basis of locally regulated rates of cell proliferation. PMID- 10367775 TI - Enzyme replacement in murine mucopolysaccharidosis type VII: neuronal and glial response to beta-glucuronidase requires early initiation of enzyme replacement therapy. AB - We have previously shown that mucopolysaccharidosis type VII (MPS VII) mice receiving six weekly injections of recombinant beta-glucuronidase from birth had improved cognitive ability and reduced central nervous system lysosomal storage. However, a single beta-glucuronidase injection at 5 wk of age did not correct neuronal storage. We define the age at which central nervous system storage in MPS VII mice becomes resistant to beta-glucuronidase therapy and determine the effect of enzyme on other tissues by comparing the histology of mice begun on therapy at various times after birth. MPS VII mice received injections on the day of birth and then weekly for 5 wk with 16,000U/g beta-glucuronidase had reduced lysosomal storage in brain. The same therapy begun on d 14 of life or thereafter failed to correct neuronal storage, even when treatment was continued for six doses. Glial responsiveness or accessibility to enzyme also depended on early treatment. In contrast, leptomeningeal, osteoblast, and retinal pigment epithelial storage reduction depended on enzyme dose rather than age at initiation of therapy. Fixed tissue macrophage storage was reduced in all treated MPS VII mice, even those receiving a single dose. These observations indicate that fixed tissue macrophages in MPS VII mice remain sensitive to enzyme replacement therapy well into adulthood although neurons are responsive or accessible to enzyme therapy early in life. Because early initiation of enzyme replacement is important to achieve a central nervous system response, these studies emphasize the importance of newborn screening for lysosomal storage diseases so that early treatment can maximize the likelihood of a favorable therapeutic response. PMID- 10367777 TI - Smooth muscle cell hypertrophy versus hyperplasia in infantile hypertrophic pyloric stenosis. AB - Infantile hypertrophic pyloric stenosis (IHPS) is characterized by hypertrophy of the pyloric muscle. It is not clearly understood whether pyloric muscle enlargement is due to hypertrophy (increase in cell size) or hyperplasia (increase in cell number). In the present study, we investigated proliferative activity as well as size and number of smooth muscle cells to understand the mechanism of pyloric muscle enlargement in IHPS. Full thickness muscle biopsy specimens were obtained from 18 IHPS patients at pyloromyotomy and from 11 age matched controls. Formalin-fixed paraffin sections were immunostained with MAb MIB-1, which stains cells in the proliferating phase of the cell cycle. The proliferative index (PI) was calculated as the percentage of positive cell nuclei. Smooth muscle cell number per bundle and cell size were measured with an image analyzer. The mean PI in IHPS (9.6 +/- 5.7%) was significantly higher than that of controls (1.3 +/- 1.2%) (p < 0.01). There was a significant inverse correlation between PI and age at operation. Smooth muscle cell number per bundle in IHPS (240.6 +/- 129.4) was significantly greater than that of the controls (134.1 +/- 49.8) (p < 0.05). Smooth muscle cell size in IHPS (298.5 +/- 59.0 microm2) was also significantly greater than that of controls (154.3 +/- 21.5 microm2) (p < 0.01). Our findings suggest that hypertrophy-and hyperplasia as well-play important roles in increasing pyloric smooth muscle mass in IHPS. PMID- 10367778 TI - Interleukin-12 in human milk. AB - The presence of interleukin-12 (IL-12) in 39 samples of human milk was investigated using the enzyme-linked immunosorbent assay. IL-12 (>40 pg/mL) was detected in 24 of the 39 samples collected (1408 +/- 2256 pg/mL, mean +/- SD, n = 24). A range of concentrations of IL-12 was observed in colostrum, transitional, and mature milk, with an apparent decrease in the mean concentration over time postpartum. PMID- 10367779 TI - Assimilation of [57Co]-labeled cobalamin in human fetal gastrointestinal xenografts into nude mice. AB - Cobalamin (Cbl) and its Cbl-binding proteins are present in amniotic fluid. Because amniotic fluid is swallowed by the embryo-fetus, we studied the ability of Cbl to be transported and metabolized across the embryo-fetal digestive tract. Human embryonic stomachs and intestines were transplanted into nude mice. The basal secretion of Cbl-binding proteins was studied by gel filtration of the graft juices. Intrinsic factor (IF) was looked for in gastric mucosa by immunohistochemistry. The uptake of [57Co]-labeled Cbl by the intestinal graft was studied by Schilling tests and HPLC. IF, haptocorrin, and a transcobalamin like protein were detected in gastric juice, with concentration ranges of 5.0 26.4, 1.9-27.1, and 5.2-12.6 pmol/mL, respectively. The IF [57Co]Cbl complex had a single isoprotein with a pI at 5.6, which was maintained after incubation with neuraminidase. Urine excretion percentages (Schilling tests) ranged from 5.5 to 21.2% and from 0.3 to 1.6% when cyano-[57Co]Cbl-IF or cyano-[57Co]Cbl, respectively, was instilled in intestinal grafts. Chloroquine reduced significantly the percentage of excreted [57Co]Cbl. The [57Co]Cbl was mainly excreted as cyano-[57Co]Cbl in urines, showing a low coenzyme conversion. In conclusion, IF is secreted by the nonstimulated embryonic stomach and lacks sialic acid. Cbl binds to it and is subsequently transported across the xenografted embryo-fetal intestine. This suggests that amniotic fluid may contribute to Cbl delivery to the embryo-fetus. PMID- 10367780 TI - L-selectin expression enhances clonogenesis of CD34+ cord blood progenitors. AB - L-selectin, a surface adhesion glycoprotein expressed on leukocytes, has a well established role in mediating inflammation and lymphocyte recirculation. Recent evidence suggests that L-selectin may also influence hematopoiesis. We observed that a greater proportion of CD34+ cells express L-selectin in cord blood compared with adult bone marrow, and we hypothesized that L-selectin expression is associated with enhanced clonogenic properties. To test this, we compared CD34+/L-selectin+ cells with CD34+/L-selectin- cells in hematopoietic clonogenic assays. From CD34+/L-selectin+ cell cultures, we observed a 3-fold increase of d 12-14 colony-forming unit-granulocyte/macrophage and multipotent progenitor cells, and a 5-fold enhancement of primitive d 21 high proliferative potential colony-forming cells compared with the progeny of CD34+/L-selectin- cells. We conclude that CD34+ cord blood cells expressing L-selectin are enriched in their clonogenic activity compared with cell fractions lacking L-selectin expression. PMID- 10367781 TI - Neutrophils from term and preterm newborn infants express the high affinity Fcgamma-receptor I (CD64) during bacterial infections. AB - The high affinity Fcgamma-receptor I (FcgammaRI, CD64) is normally expressed only to a very low extent by neutrophils. During bacterial infections, however, neutrophils from adult patients significantly increase their expression of FcgammaRI. Stimulation through FcgammaRI is a highly effective way to improve various aspects of neutrophil function, including phagocytosis. In our study the expression of FcgammaRI on neutrophils from preterm (n = 9) and term (n = 3) newborn infants, children (n = 14), and adults (n = 6) during the early phase of an acute bacterial infection was investigated. Our results showed that neutrophils from newborn infants with bacterial infection expressed FcgammaRI to a significantly higher extent than both noninfected preterm (p < 0.001) and term (p < 0.001) newborn infants and that neutrophils from preterm neonates expressed FcgammaRI to the same extent as neutrophils from term neonates and older infants, children, and adults. No difference in the neutrophil cell surface expression of FcgammaRI during bacterial infections was found among newborn infants, children, and adults. Expression of FcgammaRI probably represents an important mechanism to improve neutrophil phagocytosis as well as other aspects of neutrophil function during bacterial infections, especially in preterm infants. Our study indicates that measurement of cell surface expression of FcgammaRI on neutrophils could be a useful indicator of severe bacterial infections in preterm and term neonates, as well as in older children and adults. PMID- 10367782 TI - Heparin-binding angiogenic factors (basic fibroblast growth factor and vascular endothelial growth factor) in early neonatal life. AB - This study investigated whether serum levels of the potent angiogenic factors basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), which are abundantly produced in utero by the placenta and fetal tissues, change after birth at term, consequent to diminished angiogenic but increased adaptational demands in extrauterine life. Moreover, whether serum levels of the above factors correlate with sex, birth weight, or mode of delivery was also evaluated. One milliliter of blood was drawn from 30 healthy, appropriate for gestational age, full-term infants on d 1 (N1) and 4 (N4) postnatally. In 10 of the above cases maternal and umbilical cord blood samples were also drawn. Serum was analyzed by enzyme immunoassays, using commercial kits. Levels of bFGF and VEGF were significantly lower in maternal serum than in umbilical cord (p = 0.02 and 0.036, respectively) or N1 (p = 0.009 and 0.006, respectively) and N4 serum (p = 0.009 and 0.006, respectively). Levels of bFGF in umbilical cord serum did not differ significantly from those in N1 and N4. In contrast, levels of VEGF rose in N1, differing significantly from levels in umbilical cord serum (p = 0.008). Both factors did not change from N1 to N4. Neither bFGF nor VEGF serum levels depended on sex, mode of delivery, or birth weight. In conclusion, bFGF levels in neonates do not differ from levels in fetuses, possibly reflecting diminished angiogenesis in extrauterine life, which already has started in utero. On the contrary, neonatal levels of VEGF rise significantly after birth, possibly signifying adaptation demands, in addition to angiogenesis, as VEGF is also considered a regulator of normal function. PMID- 10367783 TI - A convenient diagnostic function test of peripheral blood neutrophils in glycogen storage disease type Ib. AB - Neutrophils from patients suffering from glycogen storage disease type Ib (GSD Ib) show several defects. one of which is a decreased rate of glucose utilization. In this study, we established experimental conditions to show the stimulation of the neutrophil respiratory burst by extracellular glucose. With phorbol-myristate-acetate as stimulus of the burst, the activity of the NADPH oxidase in GSD-Ib neutrophils hardly increased on addition of glucose. In control and GSD-type Ia neutrophils, a clear increase was observed. The lack of response to extracellular glucose in GSD-Ib neutrophils is correlated with the inability to raise intracellular glucose-6-P levels on glucose addition, thereby limiting the activity of the generation of NADPH in the hexose-monophosphate shunt. Our study shows the usefulness of this test for the diagnosis of neutrophil function abnormality in GSD-Ib patients. PMID- 10367784 TI - Outcome of Chinese patients with chronic myeloid leukaemia (CML) underwent allogeneic bone-marrow transplantation (BMT). AB - Clinical studies have shown that patients with chronic myeloid leukaemia (CML) treated with allogeneic bone-marrow transplantation (BMT) experience not only prolonged disease-free survival but also complete cure in some. Therefore, we followed a cohort of 81 Chinese patients who received allogeneic BMT. PATIENTS AND METHODS: The donors were either relatives (65 siblings, 1 parent) or unrelated volunteers (15). BMT was performed at a median interval of 11.6 months from diagnosis of CML, and the stages of disease before BMT were: first chronic phase (60 patients), accelerated or second chronic phase in (10 patients), and blastic crisis (11 patients). Three conditioning regimens were employed: Bu-Cy, Cy-TBI, or Bu-Cy-TBI. Standard cyclosporin and short methotrexate protocol were used for acute graft-versus-host disease (GvHD) prophylaxis. RESULTS: There were five graft failures with three after related BMT. Patients after related or unrelated BMT had a comparable rate of neutrophil recovery (median = 22 days), but significant delay in platelet recovery occurred after unrelated BMT (median = 34 vs. 20 days, P < 0.05). The latter also had higher incidence of acute GvHD (73% vs. 41%, P < 0.05), although the incidence of chronic GvHD was not different between groups. At a median follow-up of 43.5 months, patients after related BMT had a significantly better rate of disease-free survival (68% vs. 37.3%, P < 0.05) and overall survival (81% vs. 38.9%, P < 0.05) at 4 years. Subgroup analysis of patients after related BMT showed the outcome was better when they were transplanted at first chronic phase. Multivariate analysis showed that advanced disease (RR = 2.01, 95% CI = 1.48-2.73) significantly worsened the outcome of BMT, whereas the presence of chronic GvHD had a protective effect against relapse and survival (RR = 0.09, 95% CI = 0.02-0.38). CONCLUSION: Allogeneic BMT is a curative form of treatment for patients with CML. Treatment outcome is best for those who undergo transplants from HLA-matched siblings during the first chronic phase. PMID- 10367786 TI - Is it necessary to test patients with immune thrombocytopenic purpura (ITP) for seropositivity to HTLV-1? AB - HTLV-111 (HIV-1) has been shown to be associated with thrombocytopenia of a type resembling immune thrombocytopenic purpura (ITP). HTLV-1 is a retrovirus similar to HIV-I (HTLV-III) in a number of features, such as CD4 tropism. It is responsible for several clinical entities, including adult T-cell leukemia/lymphoma. The relationship, if any, of HTLV-1 and thrombocytopenia has not been systematically studied. To determine how frequently ITP patients are commonly infected with HTLV-1, the following study was performed. Frozen serum samples from 123 randomly selected patients with ITP were thawed and tested for antibodies to HTLV-1 by enzyme-linked immunoabsorbent assay. Positives were confirmed by Western blot. Three patients were initially found to be positive for HTLV-1. One was a female of Caribbean ancestry, one was a male HIV-1+ patient, and one was an adolescent female with no known risk factors for HIV-1. The two females later tested negative for HTLV-1. As a screening program for HTLV-1 antibodies was not introduced into blood banks until November 1988, there may have been passive transfer of the virus from intravenous immunoglobulin that these patients had received. This study of a large number of ITP patients shows that it is extremely unlikely that they are infected with HTLV-1, and, therefore, it is unnecessary to screen ITP patients for seropositivity to HTLV-1. PMID- 10367785 TI - Apoptotic effects of heparin on lymphoblasts, neutrophils, and mononuclear cells: results of a preliminary in vitro study. AB - In this study the apoptotic effects of heparin on lymphoblasts, neutrophils, and mononuclear cells were evaluated by flow cytometry for detection of sub-G1 peak, in vitro. Ten children with acute lymphoblastic leukemia (ALL) at diagnosis (Group I), six children with ALL at relapse (Group II), and 10 healthy children (controls) were included in this study. Lymphoblasts in ALL patients, and neutrophils and mononuclear cells in controls, were incubated in increasing heparin concentrations (0, 5, 10, 20 U/ml). Flow cytometric analyses were performed at 0, 1, and 2 hours of incubation in heparin for determination of the apoptotic effects of heparin. In Group I apoptosis was detected in all different levels of heparin concentration except 0 U/ml at 0, 1, and 2 hours. The apoptotic effects of heparin on blast cells peaked at the first hour in 5-, 10-, and 20 U/ml heparin concentrations (p < 0.0001). In Group II similar findings were observed only at zero hour and apoptosis was higher than those in Group I except in 5-U/ml heparin concentration (p < 0.001). Apoptosis was found to increase with heparin levels in both groups (p < 0.02). In the control group, apoptosis was detected only at the 20-U/ml heparin concentration and only at the first and second hours. Lymphoblasts are more sensitive to apoptotic effects of heparin than either neutrophils and mononuclear cells (p < 0.004). It can be suggested that low-dose heparin may cause significant apoptosis of lymphoblasts while inducing no apoptosis on neutrophils and mononuclear cells. The findings of this preliminary study indicate that further and more comprehensive research on the apoptotic effect of heparin on lymphoblasts should be done. PMID- 10367787 TI - Comparative response to splenectomy in Coombs-positive autoimmune hemolytic anemia with or without associated disease. AB - We reviewed our experience in 30 patients with direct Coombs-positive (DAT+) autoimmune hemolytic anemia (AHA) who underwent splenectomy. Twelve patients had idiopathic "warm" AHA (group I) and 18 had AHA associated with systemic diseases (group II). Complete response to splenectomy was defined as having normal hemoglobin and reticulocyte count lasting for at least 6 months without subsequent medical therapy. Subnormal but greater than 50% improvement in these parameters with or without medical therapy was considered to be a partial response. Median age was 64 (23-81) in group I and 68 (23-76) in group II. Median follow-up duration was 18 and 10.9 months, respectively. Nine of 11 (82%) evaluable patients with idiopathic AHA and 3 of 16 (19%) patients with associated disease achieved a complete response. Partial response was obtained in 2 (18%) and 6 (37%) patients in groups I and II, respectively. Both complete-response and overall-response rates were statistically different between two groups (P = 0.001 and 0.02). Postoperative courses of group I patients were uneventful except for one who developed a subphrenic abscess. Five patients in group II developed bacterial infections, which were mostly pneumonias. Our findings indicate that splenectomy is an effective treatment approach with low morbidity and mortality in patients with refractory idiopathic AHA. It should, however, be considered cautiously in AHA patients with underlying systemic diseases because of its decreased efficacy and increased surgical morbidity in this subgroup. PMID- 10367788 TI - Platelet function abnormalities in Gaucher disease patients. AB - Bleeding manifestations are common in Gaucher disease patients. Although usually attributed to thrombocytopenia, some patients with relatively high platelet counts and normal coagulation tests have hemorrhagic phenomena. To investigate whether perturbed platelet function could explain these bleeding manifestations we performed platelet aggregation tests on 32 type I adult Gaucher patients who were not severely thrombocytopenic (platelet counts >50 x 10(9)/L). Seven patients (22%) had abnormal platelet aggregation. In five, platelet aggregation was markedly reduced in response to collagen and ADP and virtually absent in response to epinephrine, whereas two patients had isolated severely impaired epinephrine-induced aggregation. In one patient platelet aggregation markedly improved following one year of enzyme replacement therapy. Incubating normal platelets with high concentrations of glucocerebroside did not impair their ability to aggregate, suggesting that plasma glucocerebroside does not directly interfere with platelet function. Platelet dysfunction is a hitherto unrecognised, relatively common cause of excessive bleeding in Gaucher patients. PMID- 10367789 TI - Effect of recombinant human granulocyte macrophage-colony stimulating factor in long-term marrow cultures from patients with aplastic anemia. AB - The hematopoietic system in patients with aplastic anemia (AA) shows both quantitative and qualitative deficiencies, i.e., reduced numbers of hematopoietic progenitor cells (HPC) and impaired HPC proliferation in long-term marrow cultures (LTMC). Since recombinant human granulocyte macrophage-colony stimulating factor (rhGM-CSF) has been shown to be a potent stimulator of normal hematopoiesis, both in vivo and in vitro, in the present study we wanted to assess the possibility of stimulating hematopoiesis in LTMC from 17 patients with AA, by weekly addition of rhGM-CSF (10 ng/ml). In LTMC from 11 patients (group of responders), rhGM-CSF induced a significant increase (4.8-fold, compared with untreated cultures) in the levels of myeloid progenitor cells; in contrast, in six patients (group of nonresponders), myeloid progenitors were refractory to this cytokine. In the group of responders, rhGM-CSF also induced a pronounced increment in the levels of nonadherent and adherent cells (5.99- and 5.18-fold, respectively, compared with untreated cultures). Among the different myelopoietic lineages, rhGM-CSF preferentially stimulated the macrophagic lineage; this was evident both at the progenitor and mature cell levels. Interestingly, the effect of rhGM-CSF in LTMC from AA patients was only transient. Indeed, the effects mentioned above were observed only during the first three weeks of culture; afterwards, myeloid progenitor and nonadherent cell levels in treated cultures declined, practically reaching the levels observed in untreated cultures. At the moment, we do not know whether this transient stimulatory effect is due to the production of inhibitory cytokines, by macrophages generated in response to rhGM CSF, or to the exhaustion of the HPC pool in AA cultures. In all 17 patients, rhGM-CSF had no effect on the kinetics of erythroid or multipotent progenitor cells. These results are in keeping with clinical studies in which it has been observed that most AA patients treated with rhGM-CSF show increments in circulating monocytes and granulocytes, as well as in bone marrow cellularity. However, little or no effect is observed on erythropoiesis. The actual mechanisms involved in the in vitro effects of rhGM-CSF on myeloid progenitor cells from AA bone marrow are still not completely understood. Future studies on this issue should be encouraged, since they may help to understand the in vivo (clinical) effects of this cytokine. PMID- 10367790 TI - Phase II trial of high-dose dexamethasone for previously treated immunoglobulin light-chain amyloidosis. AB - Immunoglobulin light-chain amyloidosis (AL) is a rare disorder characterized by production of a monoclonal light chain. This insoluble light chain, or a fragment thereof, deposits in tissues as amyloid and results in disruption of organ function and, ultimately, in death. Although melphalan and prednisone are beneficial in approximately 30% of patients with the disease, many patients fail to respond, and the median survival with this disease remains < 2 years. There is a need for new agents for those patients who fail to respond to melphalan-based chemotherapy. A study was undertaken of high-dose dexamethasone in the treatment of 19 patients with AL because of reports of its benefits in previously untreated patients with amyloidosis and its known activity in the management of multiple myeloma, which has many characteristics in common with AL. In this cohort, 3 of 19 patients showed an objective organ response of the disease. The median survival of the entire group was 11.2 months. We conclude that high-dose dexamethasone therapy is of occasional benefit in patients previously treated for amyloidosis. Combining dexamethasone with other therapies may increase the response rate. PMID- 10367791 TI - Potential of denaturing gradient gel electrophoresis for scanning of beta thalassemia mutations in India. AB - Over the last few years, substantial progress has been made in developing strategies for the detection and characterization of various mutations causing beta-thalassemia. The Indian population comprises of numerous endogamous caste groups and beta-thalassemia is seen in almost all of them. Knowledge of the spectrum of beta-thalassemia mutations in the population is a prerequisite for successful implementation of a prevention programme. Among the different approaches available today, Denaturing Gradient Gel Electrophoresis (DGGE) offers a valid technical approach which is applicable for screening of known mutants and polymorphisms as well as in locating regions of DNA bearing unknown mutations. We analysed 356 unrelated beta-thalassemia heterozygotes by DGGE and detected 30 anomalous DGGE patterns. Fifteen mutations were characterized after sequencing 25 anomalous patterns. Of these, codon 10(GCC --> GCA) is a recently reported novel beta-thalassemia mutation while -28(A --> G) and codon 121(G --> T) are being reported for the first time in the Indian population. HbS and HbE also showed two anomalous DGGE patterns each. Framework (FW) linkage studies showed that four mutations were associated with different beta-globin gene frameworks. Linkage of IVSI-5(G --> C) and cap site +1(A --> C) to FW2 and 619-bp deletion to FW1 is being observed for the first time. Multiple DGGE patterns corresponding to the same mutation is one of the major drawbacks of this technique. In spite of this, if sufficient preliminary work has been carried out to compile a comprehensive catalogue of DGGE patterns; this is a powerful approach to characterize the mutation or to localize a small region of DNA in the case of rarer mutations. PMID- 10367792 TI - Case of granulocyte colony-stimulating factor-induced Sweet's syndrome. AB - A 33-year-old male was referred with a two-week history of fevers to 40 degrees C and painful, erythematous skin and oral mucosal eruptions that had failed to respond to multiple anti-infectious agents. He had a recent diagnosis of a "myeloproliferative disorder with myelodysplastic features" on bone marrow biopsy, with associated pancytopenia. Two weeks before admission, he had been treated with a course of granulocyte colony-stimulating factor (G-CSF) at a dose of 300 microg/day in an attempt to improve his neutropenia. After four days of treatment, the fever and lesions developed. Infectious evaluation was negative; however, biopsies of the skin and oral mucosal lesions revealed histology consistent with Sweet's syndrome. Intravenous methylprednisolone (30 mg/day) was started with prompt defervescence and resolution of the lesions within days. With the increasing use of G-CSF, Sweet's syndrome is becoming more commonly recognized as an adverse effect. This is the first case of G-CSF-induced Sweet's syndrome to demonstrate gingival involvement. PMID- 10367793 TI - Interferon treatment of chronic active hepatitis C during therapy of acute lymphoblastic leukemia. AB - Due to concerns that antineoplastic therapy produces prolonged decrease in immune function, interferon treatment of chronic active hepatitis C (CAHC) has been used only at one year or longer after the end of cancer therapy. We report the experience of an 11-year-old who developed symptomatic CAHC at the start of maintenance therapy for testicular relapse of acute lymphoblastic leukemia (ALL). Significant dose reduction of maintenance therapy did not improve the tolerance of antileukemic treatment. In an effort to improve his liver disease and to deliver effective antileukemic therapy, interferon alpha and an alternative maintenance therapy regimen for ALL were initiated. The patient tolerated the combined therapy well. Interferon therapy was continued for 27 months, which was three months from the end of antineoplastic therapy. At that time serum transaminase values were normal, and no HCV viral genome was detectable. Viral genome was detected 10 months later. The combined effects of interferon and antineoplastic therapy resulted in myelosuppression requiring dose reduction of both treatments. The patient remains asymptomatic and with no evidence of recurrent leukemia more than six years from diagnosis of relapse. The effect on the status of this patient's CAHC was similar to that reported among leukemic patients who underwent an interferon course more than one year from the end of antineoplastic therapy. Interferon treatment of CAHC can be given concomitantly with antineoplastic therapy. PMID- 10367794 TI - Fatal virus-associated hemophagocytic syndrome associated with coexistent chronic active hepatitis B and acute hepatitis C virus infection. AB - A 28-year-old man was admitted to our department with intermittent fever, hepatosplenomegaly and pancytopenia. Liver parameters and serum ferritin were markedly elevated. Bone marrow biopsy showed hypocellularity, histiocytic hyperplasia, and hemophagocytosis consistent with a virus-associated hemophagocytic syndrome (VAHS). There was serological evidence of chronic active hepatitis B and acute hepatitis C virus infection. The patient died despite aggressive immunosuppressive and supportive treatment. Autopsy revealed signs of acute viral hepatitis with cholestasis. Histiocytes engaged in hemophagocytosis were observed in bone marrow and spleen. The condition was interpreted as VAHS associated with chronic active hepatitis B and acute hepatitis C virus infection. To our knowledge this is the first report of a hemophagocytic syndrome in that setting. PMID- 10367795 TI - Fetal hemoglobin expression in the compound heterozygous state for -117 (G-->A) Agamma HPFH and IVSII-745 (C-->G) beta+ thalassemia: a case study. AB - We studied a family in which two inherited defects of the non-alpha-globin cluster segregate: Greek hereditary persistence of fetal hemoglobin (HPFH) and beta-thalassemia. The compound heterozygote is a healthy man with 43% HbF, Ggamma/Agamma ratio (27:73) differing from that of 10 simple heterozygotes for the Greek HPFH (92:8), normal levels of total Hb (13.3 g/dl), and reduced HbA2 levels comparing with the levels of beta-thal heterozygotes for the same mutation. Molecular analysis of the beta-globin genotype revealed the presence of the IVSII-745 (C-->G) beta+ RNA splice mutation in trans with the -117 G-->A Greek HPFH. The beta+ mutation was linked to haplotype VII and the Greek HPFH was associated with haplotype Ia. The father of the compound heterozygote carries the Greek HPFH in trans with the -158 C-->T on the Ggamma promoter, which is linked with haplotype IV. He presented 13.5% HbF with a Ggamma/Agamma ratio 75:25. His daughter was a compound heterozygote for the IVSII-745 mutation in trans with the -158 C-->T, while her HbF levels were 3.7% with a Ggamma/Agamma ratio 31:69. PMID- 10367796 TI - Spontaneous regression associated with apoptosis in a patient with acute-type adult T-cell leukemia. AB - We describe a 76-year-old man with acute-type adult T-cell leukemia, who demonstrated a spontaneous decrease in leukemic cell number, apparently coincident with apoptotic cell death. On admission the patient's white blood cell count was 38.9 x 10(9)/l with 77% abnormal lymphocytes. He also had hypoproteinemia (4.3 g/dl) from protein losing enteropathy. After admission the leukemic cell count decreased without chemotherapy, reaching 5.9 x 10(9)/l after 2 months. Studies of peripheral lymphocytes demonstrated appearance of the apoptotic cells and DNA ladder formation from the beginning of regression. Same truncated proviral DNA was recognized in primary ATL cells through the whole clinical course. The hypoproteinemia improved with intravenous nutrition, followed by increase of the leukemic cells. This case is the first report that demonstrates tumor-cell apoptosis induced clinical regression in adult T-cell leukemia. Further, we speculate that the hypoproteinemia may have been involved in the leukemic cell apoptosis. PMID- 10367797 TI - Distinctive AgNOR patterns of myeloid and lymphoid blasts in acute leukemia. AB - Bone-marrow aspiration smears of 38 cases of acute leukemia (19 acute lymphoblastic leukemia, 19 acute nonlymphocytic leukemia) were stained for argyrophilic nucleolar organiser regions (AgNOR). AgNOR were assessed numerically and morphologically. There were highly significant differences in AgNOR morphology between acute lymphoblastic leukemia and acute nonlymphocytic leukemia ANLL: lymphoblast AgNORs were usually small (<3 microm) "dots/chips," whereas myeloblasts showed larger "blebs," or a combination of the two types (complex structures). AgNOR number was significantly less in acute lymphoblastic leukemia. In acute nonlymphocytic leukemia cases, AgNOR number also showed direct correlation with Ki67 reactivity of leukemic blasts. PMID- 10367798 TI - Normal binding of plasma von Willebrand factor to platelets in essential thrombocythemia. PMID- 10367799 TI - Multifocal T-cell lymphoma of bone. PMID- 10367800 TI - Androgen-induced erythrocytosis: is it erythropoietin? PMID- 10367801 TI - Thiamine deficiency in a patient receiving chemotherapy for acute myeloblastic leukemia. PMID- 10367802 TI - Iron deficiency anemia of unknown etiology and/or resistance to the treatment: the sole manifestation of adult celiac disease (CD) PMID- 10367803 TI - Remission of pure-red-cell aplasia associated with operative cure of lung cancer. PMID- 10367805 TI - Improved stroke imaging techniques. PMID- 10367804 TI - Buckle up! Is not enough: enhancing protection of the restrained child. PMID- 10367806 TI - New pharmaceutical approaches to stroke prevention, treatment. PMID- 10367807 TI - Care gap "unconscionable"; universal coverage AAP aim. American Academy of Pediatrics. PMID- 10367808 TI - From the Centers for Disease Control and Prevention. Motor-vehicle safety: a 20th century public health achievement. PMID- 10367809 TI - Cost-effectiveness of interferon treatment for hepatitis C. PMID- 10367810 TI - Cost-effectiveness of interferon treatment for hepatitis C. PMID- 10367811 TI - Epidural analgesia and cesarean delivery. PMID- 10367812 TI - Epidural analgesia and cesarean delivery. PMID- 10367813 TI - Epidural analgesia and cesarean delivery. PMID- 10367814 TI - Yoga for carpal tunnel syndrome. PMID- 10367815 TI - Yoga for carpal tunnel syndrome. PMID- 10367816 TI - Yoga for carpal tunnel syndrome. PMID- 10367817 TI - Yoga for carpal tunnel syndrome. PMID- 10367818 TI - The realities of resident research requirements. PMID- 10367819 TI - Hormone replacement therapy and risk of breast cancer with a favorable histology: results of the Iowa Women's Health Study. AB - CONTEXT: Long-term, postmenopausal use of hormone replacement therapy (HRT) appears to increase breast cancer risk. Whether the effect of HRT use on risk of breast cancer varies among histological types of invasive carcinoma is unknown. OBJECTIVE: To determine associations between HRT use and risk of ductal carcinoma in situ (DCIS), invasive carcinoma with favorable histology, and invasive ductal or lobular carcinoma. DESIGN: Prospective cohort study whose participants were followed up continuously for 11 years from January 1986 to December 1996. SETTING AND PARTICIPANTS: Iowa Women's Health Study, a population-based random sample of postmenopausal women aged 55 to 69 years in 1986. A total of 1520 incident breast cancer cases occurred in the at-risk cohort of 37 105 women. MAIN OUTCOME MEASURES: Multivariate-adjusted relative risk (RR) of tumor-specific breast cancers associated with duration of ever, current, or past HRT use. RESULTS: Duration of ever HRT use was associated with risk of invasive carcinoma with a favorable histology, with an RR of 1.81 (95% confidence interval [CI], 1.07-3.07) for those who used HRT 5 or fewer years vs an RR of 2.65 (95% CI, 1.34-5.23) for those who used HRT for more than 5 years (P for trend = .005) after adjustment for age and other breast cancer risk factors. There was no association between ever HRT use and the incidence of DCIS or invasive ductal or lobular carcinoma. Among current hormone users, after adjusting for age and other breast cancer risk factors, the RRs (95% CIs) of invasive carcinoma with a favorable histology were 4.42 (2.00-9.76) and 2.63 (1.18-5.89) for 5 or fewer years of use and for more than 5 years of use, respectively. Risk of invasive ductal or lobular carcinoma was associated with current use (< or =5 years) of HRT with an RR of 1.38 (95% CI, 1.03-1.85). CONCLUSIONS: Exposure to HRT was associated most strongly with an increased risk of invasive breast cancer with a favorable prognosis. These data add important clinical information for assessing the risks and benefits of HRT use. PMID- 10367820 TI - The unreliability of individual physician "report cards" for assessing the costs and quality of care of a chronic disease. AB - CONTEXT: Physician profiling is widely used by many health care systems, but little is known about the reliability of commonly used profiling systems. OBJECTIVES: To determine the reliability of a set of physician performance measures for diabetes care, one of the most common conditions in medical practice, and to examine whether physicians could substantially improve their profiles by preferential patient selection. DESIGN AND SETTING: Cohort study performed from 1990 to 1993 at 3 geographically and organizationally diverse sites, including a large staff-model health maintenance organization, an urban university teaching clinic, and a group of private-practice physicians in an urban area. PARTICIPANTS: A total of 3642 patients with type 2 diabetes cared for by 232 different physicians. MAIN OUTCOME MEASURES: Physician profiles for their patients' hospitalization and clinic visit rates, total laboratory resource utilization rate and level of glycemic control by average hemoglobin A1c level with and without detailed case-mix adjustment. RESULTS: For profiles based on hospitalization rates, visit rates, laboratory utilization rates, and glycemic control, 4% or less of the overall variance was attributable to differences in physician practice and the reliability of the median physician's case-mix adjusted profile was never better than 0.40. At this low level of physician effect, a physician would need to have more than 100 patients with diabetes in a panel for profiles to have a reliability of 0.80 or better (while more than 90% of all primary care physicians at the health maintenance organization had fewer than 60 patients with diabetes). For profiles of glycemic control, high outlier physicians could dramatically improve their physician profile simply by pruning from their panel the 1 to 3 patients with the highest hemoglobin A1c levels during the prior year. This advantage from gaming could not be prevented by even detailed case-mix adjustment. CONCLUSIONS: Physician "report cards" for diabetes, one of the highest-prevalence conditions in medical practice, were unable to detect reliably true practice differences within the 3 sites studied. Use of individual physician profiles may foster an environment in which physicians can most easily avoid being penalized by avoiding or deselecting patients with high prior cost, poor adherence, or response to treatments. PMID- 10367821 TI - A prospective study of coffee consumption and the risk of symptomatic gallstone disease in men. AB - CONTEXT: Coffee has several metabolic effects that could reduce the risk of gallstone formation. OBJECTIVE: To examine the association between coffee consumption and the risk of symptomatic gallstone disease in men. DESIGN AND SETTING: The Health Professionals Follow-up Study, a prospective cohort study, in which the consumption of coffee and other caffeinated drinks was assessed starting in 1986 as part of the 131-item food frequency questionnaire given to US male health professionals with follow-up through 1996. PARTICIPANTS: A total of 46008 men, aged 40 to 75 years in 1986, without history of gallstone disease. MAIN OUTCOME MEASURES: Newly symptomatic gallstone disease (diagnosed by ultrasonography or x-ray) or a cholecystectomy. RESULTS: During 404 166 person years of follow-up, 1081 subjects reported symptomatic gallstone disease, of whom 885 required cholecystectomy. After adjusting for other known or suspected risk factors, compared with men who did not consume regular coffee in 1986 and 1990, the adjusted relative risk (RR) for those who consistently drank 2 to 3 cups of regular coffee per day was 0.60 (95% confidence interval [CI], 0.42-0.86) and for those who drank 4 or more cups per day the RR was 0.55 (95% CI, 0.33-0.92). All coffee brewing methods showed a decreased risk. The risk of symptomatic gallstone disease also declined with increasing caffeine intake (P for trend = .005). After controlling for known or suspected risk factors, the RR for men in the highest category of caffeine intake (>800 mg/d) compared with men in the lowest category (< or =25 mg/d) was 0.55 (95% CI, 0.35-0.87). In contrast, decaffeinated coffee was not associated with a decreased risk. CONCLUSIONS: In this cohort of US men, coffee consumption may have helped to prevent symptomatic gallstone disease. PMID- 10367822 TI - Impairment of endothelial functions by acute hyperhomocysteinemia and reversal by antioxidant vitamins. AB - CONTEXT: Increased levels of homocysteine are associated with risk of cardiovascular disease. Homocysteine may cause this risk by impairing endothelial cell function. OBJECTIVE: To evaluate the effect of acute hyperhomocysteinemia with and without antioxidant vitamin pretreatment on cardiovascular risk factors and endothelial functions. DESIGN AND SETTING: Observer-blinded, randomized crossover study conducted at a university hospital in Italy. SUBJECTS: Twenty healthy hospital staff volunteers (10 men, 10 women) aged 25 to 45 years. INTERVENTIONS: Subjects were given each of 3 loads in random order at 1-week intervals: oral methionine, 100 mg/kg in fruit juice; the same methionine load immediately following ingestion of antioxidant vitamin E, 800 IU, and ascorbic acid, 1000 mg; and methionine-free fruit juice (placebo). Ten of the 20 subjects also ingested a placebo load with vitamins. MAIN OUTCOME MEASURES: Lipid, coagulation, glucose, and circulating adhesion molecule parameters, blood pressure, and endothelial functions as assessed by hemodynamic and rheologic responses to L-arginine, evaluated at baseline and 4 hours following ingestion of the loads. RESULTS: The oral methionine load increased mean (SD) plasma homocysteine level from 10.5 (3.8) micromol/L at baseline to 27.1 (6.7) micromol/L at 4 hours (P<.001). A similar increase was observed with the same load plus vitamins (10.0 [4.0] to 22.7 [7.8] micromol/L; P<.001) but no significant increase was observed with placebo (10.1 [3.7] to 10.4 [3.2] micromol/L; P=.75). Coagulation and circulating adhesion molecule levels significantly increased after methionine ingestion alone (P<.05) but not after placebo or methionine ingestion with vitamins. While the mean (SD) blood pressure (-7.0% [2.7%]; P<.001), platelet aggregation response to adenosine diphosphate ( 11.4% [4.5%]; P=.009) and blood viscosity (-3.0% [1.2%]; P=.04) declined in these parameters 10 minutes after an L-arginine load (3 g) following placebo, the increase after methionine alone (-2.3% [1.5%], 4.0% [3.0%], and 1.5% [1.0%], respectively; P<.05), did not occur following methionine load with vitamin pretreatment (-6.3% [2.5%], -7.9% [3.5%], and -1.5% [1.0%], respectively; P=.24). CONCLUSION: Our data suggest that mild to moderate elevations of plasma homocysteine levels in healthy subjects activate coagulation, modify the adhesive properties of endothelium, and impair the vascular responses to L-arginine. Pretreatment with antioxidant vitamin E and ascorbic acid blocks the effects of hyperhomocysteinemia, suggesting an oxidative mechanism. PMID- 10367823 TI - Inhaled budesonide in addition to oral corticosteroids to prevent asthma relapse following discharge from the emergency department: a randomized controlled trial. AB - CONTEXT: Relapses of acute asthma following emergency department (ED) discharge can be reduced with systemic corticosteroid treatment. However, whether inhaled corticosteroids (ICSs) provide additional benefit is not known. Objective To determine whether the addition of ICSs to oral corticosteroid treatment would reduce relapses in patients with acute asthma discharged from the ED. DESIGN AND SETTING: Placebo-controlled, double-blind, randomized clinical trial conducted in a community teaching hospital ED in Canada between November 1995 and September 1997, with a 21-day follow-up. PARTICIPANTS: A total of 1006 consecutive patients aged 16 to 60 years presented to the ED with acute asthma; after excluding those using oral and/or inhaled corticosteroids as well as those meeting other exclusion criteria, 188 were included in the study. INTERVENTIONS: Patients were discharged with a nontapering course of oral prednisone (50 mg/d) for 7 days. In a double-blind fashion, patients were randomly assigned to 1600 microg/d of inhaled budesonide (n = 94) or identical placebo (n = 94) for 21 days. MAIN OUTCOME MEASURES: Incidence of relapse, defined as an unscheduled visit for worsening asthma symptoms, in budesonide vs placebo groups. Secondary outcomes included response to the Asthma Quality of Life Questionnaire, beta2-agonist use, symptom score, global asthma improvement assessment, and pulmonary function. RESULTS: Five patients in the budesonide group and 3 in the placebo group either dropped out or were lost to follow-up but were included in primary analyses. After 21 days, 12 (12.8%) of 94 patients in the budesonide group experienced a relapse compared with 23 (24.5%) of 94 in the placebo group, a 48% relapse reduction (P=.049). Asthma Quality of Life Questionnaire scores were higher (better quality) in the budesonide group (P=.001), as well as for all domain scores (P=.001 to .01). Fewer beta2-agonist activations were used at the end of the trial by patients receiving budesonide (2.4/d vs 4.2/d; P=.01). Symptom scores (P=.001 to .004) and self-assessed asthma improvement scores (based on a 7 point Likert scale) (6.2 vs 5.2; P<.001) were higher (indicating fewer symptoms) for budesonide vs placebo. There were no differences in pulmonary function between the groups (peak expiratory flow rate: budesonide, 437 vs placebo, 453 L/min; P= .39) at 21 days. Using this approach, as few as 9 patients would require budesonide to prevent 1 relapse. CONCLUSIONS: Patients discharged from the ED following treatment for acute asthma benefit from added treatment with high-dose inhaled budesonide for 21 days compared with oral corticosteroids alone. PMID- 10367825 TI - The need for global action against multidrug-resistant tuberculosis. PMID- 10367824 TI - Smallpox as a biological weapon: medical and public health management. Working Group on Civilian Biodefense. AB - OBJECTIVE: To develop consensus-based recommendations for measures to be taken by medical and public health professionals following the use of smallpox as a biological weapon against a civilian population. PARTICIPANTS: The working group included 21 representatives from staff of major medical centers and research, government, military, public health, and emergency management institutions and agencies. Evidence The first author (D.A.H.) conducted a literature search in conjunction with the preparation of another publication on smallpox as well as this article. The literature identified was reviewed and opinions were sought from experts in the diagnosis and management of smallpox, including members of the working group. CONSENSUS PROCESS: The first draft of the consensus statement was a synthesis of information obtained in the evidence-gathering process. Members of the working group provided formal written comments that were incorporated into the second draft of the statement. The working group reviewed the second draft on October 30, 1998. No significant disagreements existed and comments were incorporated into a third draft. The fourth and final statement incorporates all relevant evidence obtained by the literature search in conjunction with final consensus recommendations supported by all working group members. CONCLUSIONS: Specific recommendations are made regarding smallpox vaccination, therapy, postexposure isolation and infection control, hospital epidemiology and infection control, home care, decontamination of the environment, and additional research needs. In the event of an actual release of smallpox and subsequent epidemic, early detection, isolation of infected individuals, surveillance of contacts, and a focused selective vaccination program will be the essential items of an effective control program. PMID- 10367826 TI - Hormone replacement therapy and risk of breast cancer. PMID- 10367828 TI - JAMA Patient Page: asthma. PMID- 10367827 TI - Can physician profiles be trusted? PMID- 10367829 TI - A unifying model to explain the increased incidence and higher mortality of prostate cancer in black men. PMID- 10367831 TI - Cryotherapy for small renal cell tumors: con. PMID- 10367830 TI - Cryotherapy for small renal cell tumors: pro. PMID- 10367832 TI - History of the neurobiology of the pelvis. AB - The contemporary clinical management of a variety of pelvic disorders has benefited greatly from both basic science research discoveries and clinical observations in pelvic neurobiology. The foundation of this scientific discipline emanates from historical advances in the neuroanatomy and neurochemistry of the pelvic region. These advances, many of which were controversial when introduced, have served to shape currently accepted views regarding the neuroregulatory basis of pelvic functions. This report highlights the major contributions in the history of pelvic neurobiology, providing both an overview of the evolution of scientific thought in this discipline and insight into the role of this discipline in the development and practice of medicine and surgery related to the pelvic region. PMID- 10367833 TI - Impact of drug therapy on benign prostatic hyperplasia-specific quality of life. AB - Quality of life (QOL) is an important issue when assessing medical treatment of benign prostatic hyperplasia (BPH). There are many QOL questionnaires available, and disease-specific questionnaires are being developed. Currently, most patients undergoing treatment for BPH receive alpha-blockers or finasteride. To determine which QOL measures are being used, we did a Medline search covering the past 10 years and found 11 studies in which BPH-QOL was investigated. The wide variety of questionnaires used made comparison between drug studies difficult. When comparing studies that used at least one similar questionnaire to that of another drug study, we found alpha-blocker treatment excelled over finasteride in improving BPH-QOL in the areas of earlier response, larger decreases in mean changes, and reduced sexual side effects. QOL assessment should be a routine part of BPH treatment, and more standardized and validated measures should be used to allow for comparative, meaningful analyses. PMID- 10367834 TI - Robotic-assisted laparoscopic pyeloplasty: a pilot study. AB - OBJECTIVES: Robotic technology has been employed to manipulate the laparoscope during urologic procedures. However, to our knowledge, robotic technology has not been previously applied to actually perform the urologic laparoscopic procedure. The objective of this study was to determine the feasibility and efficacy of performing robotic-assisted laparoscopic pyeloplasty and compare it with conventional laparoscopic pyeloplasty in an acute porcine model. METHODS: Five female swine (10 kidneys) were prospectively randomized to undergo unstented robotic-assisted laparoscopic pyeloplasty (6 kidneys) or conventional laparoscopic pyeloplasty (4 kidneys). Robotic pyeloplasty was performed with the Zeus robotic system, which incorporates three remote-controlled interactive arms: one voice-activated arm to control the laparoscope and two robotic arms to manipulate purpose-designed instruments. Tissue dissection and transection of ureteropelvic junction area were performed manually by conventional laparoscopy. The pyeloureteric anastomosis during the robotic-assisted pyeloplasty was performed completely robotically from a remote workstation using running 5-0 absorbable sutures. Conventional laparoscopic pyeloplasty was performed manually by laparoscopic intracorporeal suturing and knot-tying techniques. Immediate patency and anastomotic integrity were evaluated by intravenous indigo carmine and ex vivo retrograde ureteropyelogram. RESULTS: In comparing robotic and conventional laparoscopic pyeloplasty, the following data were obtained: total surgical time (115.2 versus 94.5 minutes, P = 0.2), anastomosis time (75.7 versus 64.3 minutes, P = 0.3), and total number of suture-bites per ureter (13.0 versus 12.5, P = 0.8). Anastomoses were immediately watertight in 5 of 6 robotic and 3 of 4 conventional pyeloplasties. CONCLUSIONS: Robotic-assisted laparoscopic pyeloplasty is a feasible and effective procedure that may enhance surgical dexterity and precision. This has implications for clinical applications of laparoscopic telesurgery in the future. PMID- 10367835 TI - A new temporary catheter (ContiCath) for the treatment of temporary, reversible, postoperative urinary retention. AB - OBJECTIVES: To describe the initial experience of a newly designed temporary urethral catheter, ContiCath, as an aid in the management of postoperative or temporary outflow obstruction. In patients with normal detrusor and sphincter function, this catheter allows volitional voiding while maintaining an open prostatic urethra. METHODS: In a pilot study, 64 nonconsecutive patients with postoperative or temporary urinary retention, at eight clinical trial sites, were enrolled for the placement of this temporary catheter. Three patients did not have the catheter placed because of placement failure because of either a large median lobe or a urethral stricture. The remaining 61 patients were divided into three groups: those with non-neuropathic causes of retention and retention for 1 week or less (37 patients), those with non-neuropathic causes of retention and retention for longer than 1 week (19 patients), and those with neuropathic causes of retention and retention for longer than 1 week (5 patients). The ContiCath is placed in the office setting, in the same fashion as a Foley catheter. A blue prolene tether extends from the bulbar urethra to the meatus to assist in the removal of the device. Patients were then reassessed at 3 hours, and at 7, 14, 21, and 28 days, at which point the device was removed. RESULTS: In patients with a neuropathic cause for their retention (5 patients) and those with non neuropathic causes of retention and retention for longer than 1 week (19 patients), only 3 patients were able to void after the catheter was placed. Of the 37 patients with a non-neuropathic cause and retention 1 week or less, controlled voiding was seen in 33 patients (89%). Controlled voiding was defined as the patient's volitional ability to initiate and stop his urinary stream. There were no complications with catheter placement; however, 8 patients (24.2%) had minor adverse experiences of frequency/urgency (n = 3), incontinence (n = 3), migration of the catheter (n = 1), and pain (n = 1). CONCLUSIONS: ContiCath offers an alternative to an indwelling Foley catheter in men with temporary bladder outlet obstruction and urinary retention. PMID- 10367836 TI - Extracorporeal magnetic innervation therapy for stress urinary incontinence. AB - OBJECTIVES: To report the first data from a prospective clinical study to determine the feasibility of using extracorporeal magnetic innervation (ExMI) for the treatment of stress urinary incontinence. METHODS: We studied 83 women with demonstrable stress urinary incontinence. Treatments were for 20 minutes, twice a week for 6 weeks. For treatment, the patient sits fully clothed on a special chair; within the seat is a magnetic field generator that produces the rapidly changing magnetic field flux. Objective measures included bladder diaries, dynamic pad weight testing, urodynamic studies, and quality of life survey. RESULTS: Fifty patients have been followed up for longer than 3 months (33 patients for less than 3 months); 17 patients (34%) were dry, 16 (32%) were using not more than 1 pad per day, and 17 (34%) were using more than 1 pad per day. Pad use was reduced from 2.5 to 1.3 (P = 0.001) and leak episodes per day were reduced from 3.3 to 1.7 (P = 0.001). The pad weight was reduced from 20 to 15 g. Detrusor instability was found in 5 patients before but was demonstrated in only 1 patient after treatment. CONCLUSIONS: ExMI therapy offers a new effective modality for pelvic floor muscle stimulation. ExMI is painless, there is no need for a probe, and no need to undress for treatments. Longer follow-up is required to determine how long the benefits of treatment last and whether retreatment will be necessary. PMID- 10367837 TI - Efficacy of sildenafil citrate in prostate brachytherapy patients with erectile dysfunction. AB - OBJECTIVES: To ascertain the efficacy of sildenafil citrate (Viagra) in patients with erectile dysfunction (ED) either before or after prostate brachytherapy by an open-label, nonrandomized study. METHODS: Sixty-two patients who underwent prostate brachytherapy between March 1995 and July 1998, had ED either before or after brachytherapy, and were interested in treatment with sildenafil comprised the patient population. Clinical and treatment parameters evaluated for medication efficacy included patient age at brachytherapy and at medication administration, hypertension, diabetes, smoking history, onset of ED, potency status before implant, frequency of intercourse before brachytherapy (if potent), use of neoadjuvant hormonal manipulation, use of moderate dose external beam radiation therapy before implantation, choice of isotope, V100 (the percentage of the prostate volume receiving at least 100% of the prescribed minimal peripheral dose), and sildenafil dose. RESULTS: Fifty (80.6%) of 62 patients responded favorably to sildenafil. None of the treatment parameters predicted medication failure, and among the clinical parameters, only diabetes predicted failure (3 of 5) and only with borderline statistical validity (P = 0.046). CONCLUSIONS: Our results suggest brachytherapy-induced impotence is as amenable to sildenafil treatment as ED from other causes. In addition, our 80.6% success rate is comparable to reported results for patients who underwent bilateral nerve-sparing radical prostatectomy and significantly better than patients who underwent unilateral nerve-sparing or non-nerve-sparing approaches. PMID- 10367838 TI - The Internet and patient education--resources and their reliability: focus on a select urologic topic. AB - OBJECTIVES: The information revolution triggered by the rapid growth of the Internet has allowed healthcare providers and patients to access a rapidly expanding volume of information. To address the quality of this information, a survey of the data on a single urology-related topic available on the Internet was performed. METHODS: The search on the World Wide Web (Web) was performed using the search engine HotBot and search directory Yahoo. The Web pages were assessed according to their relevancy to the topic chosen. Relevance rates were derived from the number of relevant sites divided by the total number of sites found. Relevant sites were subsequently ranked for quality on the basis of their accuracy, comprehensiveness, and objectivity. HotBot was then subsequently divided by domain, with each assessed separately. Yahoo was analyzed in its entirety. The resources were then compared for relevance and quality of information. RESULTS: When using the keyword "Viagra," HotBot responded with 15,109 hits. Yahoo presented 51 hits under the category, "Health: Pharmacy: Drugs and Medications: Specific Drugs and Medications: Viagra (Sildenafil)." The relevance rate for the first 50 hits in the search engine HotBot was 0.08. The relevance rates for the edu and org domains found by HotBot were 0.22 and 0.24, respectively; those for com and net were both 0.10. The relevance rate for the search directory Yahoo was 0.20. For relevant sites, the quality of the information presented was significantly higher in the Yahoo and in the HotBot domains hosted by nonprofit organizations when compared with HotBot in general and with its commercially oriented domains. HotBot overall was found to contain seven excellent sites, of which only three were found within Yahoo. CONCLUSIONS: Although the medical information available on the Web has proliferated at a remarkable rate, the number of Web sites providing complete, nonbiased information continues to represent only a small portion of the total. We have shown that the search directory Yahoo reduced the number of irrelevant sites significantly, but at the same time, some very valuable information available in HotBot was missing. At present, it may be useful to conduct searches within Yahoo followed by a review of both the edu and org HotBot domains. PMID- 10367839 TI - Radical nephrectomy and nephroureterectomy in the octogenarian and nonagenarian: comparison of laparoscopic and open approaches. AB - OBJECTIVES: To retrospectively compare the outcome of laparoscopic and open radical nephrectomy or nephroureterectomy in patients 80 years old or older, inasmuch as the tolerance profile of major laparoscopic renal surgery in comparison to open surgery in the elderly patient has not been previously reported. METHODS: Since September 1997, 11 patients 80 years old or older underwent retroperitoneal laparoscopic radical nephrectomy or nephroureterectomy for cancer. These patients were compared with 6 consecutive patients 80 years old or older who underwent comparable open surgery at our institution since January 1994. No tumor had computed tomographic evidence of lymphatic, vascular, or perirenal extension. RESULTS: Baseline parameters were comparable between the laparoscopic and open groups. The laparoscopic group had a similar median surgical time (210 minutes versus 175 minutes; P = 0.1) and blood loss (150 mL versus 125 mL; P = 0.8) compared with the open group. However, specimen weight was larger in the laparoscopic group (568 g versus 292 g; P = 0.04). Moreover, the laparoscopic group had a quicker resumption of oral intake (less than 1 day versus 4 days; P <0.001), decreased narcotic requirements (14 mg versus 326 mg; P = 0.004), shorter hospital stay (2 days versus 6 days; P <0.001), and faster convalescence (14 days versus 42 days; P <0.001) compared with the open group. CONCLUSIONS: Retroperitoneal laparoscopic radical nephrectomy and nephroureterectomy are well tolerated by the elderly patient. Although our sample size was small, it appears that laparoscopy is an excellent alternative to open surgery for excision of selected renal malignancies in the octogenarian and nonagenarian population. PMID- 10367840 TI - Male urethral carcinoma: analysis of treatment outcome. AB - OBJECTIVES: To evaluate our experience with primary carcinomas of the male urethra and to analyze the impact of tumor variables and treatment on overall, disease-specific, local recurrence-free, and metastasis-free survival. METHODS: Between 1958 and 1996, we identified 46 men with primary carcinoma of the bulbar and anterior urethra. The median follow-up was 125 months (1 to 336). The patients were stratified by stage, nodal status, histologic type, treatment, type of surgery, site of disease, year at diagnosis, and smoking status. RESULTS: The overall survival and disease-specific survival rates at 5 years were 42% and 50%, respectively. The recurrence-free survival and metastasis-free survival rates at 5 years were 51% and 56%, respectively. The overall survival rate was 83% for superficial disease versus 36% for invasive tumors. The overall survival rate was 26% for tumors of the bulbar urethra versus 69% for tumors of the anterior urethra. CONCLUSIONS: Current modalities of treatment are ineffective for local control and survival. New treatment strategies are needed for urethral cancer. PMID- 10367842 TI - Elevated urinary norepinephrine in interstitial cystitis. AB - OBJECTIVES: To measure urinary catecholamines and determine the extent to which they may be elevated in urine from patients with interstitial cystitis (IC). METHODS: Random urine samples from patients with IC (n = 111) and urine from normal volunteers (n = 92) were acidified on collection (voided and catheterized specimens) and assayed for catecholamine (norepinephrine or normetanephrine) by enzyme-linked immunosorbent assay. Creatinine levels in these urine samples were also measured. RESULTS: Analysis of the data indicated that patients with IC had a higher urinary level of the neurotransmitter norepinephrine compared with the measured levels in the urine of normal volunteers (89.1 +/- 58.3 versus 54.9 +/- 37.1 microg/g creatinine, P <0.05). The metabolite normetanephrine was similar in the urine samples from these two groups. Urine from patients with bladder outlet obstruction (n = 11) did not have elevated amounts of urinary norepinephrine. The norepinephrine levels were not statistically different in the urine samples from patients with symptomatic and asymptomatic IC. The elevated urinary levels in patients with IC did not decrease after treatment with sodium pentosanpolysulfate (Elmiron), heparinoids, dimethyl sulfoxide, or combinations of these during 1 to 15 months of treatment. CONCLUSIONS: Norepinephrine was found to be elevated in the urine from patients with IC compared with urine from normal controls. This would be consistent with increased sympathetic (adrenergic) activity from the bladders of patients with IC or possibly from increased adrenal activity, since stress is associated with symptom increase in some patients with IC. Norepinephrine levels did not decrease with treatment nor did they differ between symptomatic and asymptomatic patients at the time of urine collection. PMID- 10367841 TI - Natural history of interstitial cystitis in 274 patients receiving sulfated polysaccharide therapy. AB - OBJECTIVES: To assess the natural history of interstitial cystitis in the presence of sulfated polysaccharide treatment. METHODS: This was a longitudinal study of 274 patients. Questionnaires were administered at first visit to obtain information on demographic characteristics, medical history, other risk factors, and type and severity of symptoms. Follow-up questionnaires were administered at subsequent visits to measure symptom progress. Patient status over time was measured for three symptoms: pain, urgency, and nocturia. Changes in symptom and severity were assessed at 6, 12, and 24 months on treatment. Comparisons of symptom change from baseline to 6 and 12 months were assessed for different characteristics among patients with the most severe symptoms. RESULTS: After 1 year of treatment, a decrease of two or more points in symptom score was observed for 33.5% of all patients for pain and 35.4% for urgency. Among patients with the most severe symptoms, a decrease of two or more points was observed in more than 54% for pain and urgency; 55.7% experienced remission to the moderate and mild level (49.2% and 6.5%, respectively). There was no appreciable decrease in nocturia at any severity level. With the exception of feeling heavy and experiencing dull pain at baseline, patients who did not report a specific type of pain improved more than those who did. CONCLUSIONS: The results of this study suggest that treatment with sulfated polysaccharides can help alleviate the symptoms of patients suffering from the most severe stages of interstitial cystitis. PMID- 10367843 TI - Upper urinary tract tumors developing after treatment of superficial bladder cancer: 7-year follow-up of 591 consecutive patients. AB - OBJECTIVES: To evaluate upper urinary tract tumor (UUTT) incidence and characteristics in 591 consecutive patients with low-, intermediate-, or high risk superficial bladder cancer, who were followed up for at least 5 years or until death. METHODS: From 1986 to 1992, 591 patients were treated for superficial bladder cancer: 216 patients with primary, solitary, low-grade (G1 G2), and low-stage (Ta-T1) superficial bladder cancer were considered at low risk of disease recurrence and treated with transurethral resection (TUR) alone; 182 patients with recurrent or multifocal superficial bladder cancer were considered at intermediate risk of disease recurrence or progression and treated with intravesical chemotherapy after TUR; 193 patients with carcinoma in situ, high grade (G3) superficial bladder tumor, or intravesical chemotherapy failure were considered at high risk of disease recurrence or progression and treated with bacille Calmette-Guerin (BCG). RESULTS: After a median follow-up of 86 months (range 20 to 143), 2 (0.9%) of 216 patients at low risk, 4 (2.2%) of 182 patients at intermediate risk, and 19 (9.8%) of 193 patients at high risk developed UUTTs. The incidence of UUTTs is significantly higher in patients at high risk than in those at low risk (P = 0.0004, odds ratio = 11.6, 95% confidence interval [CI] 2.5 to 40.7) or at intermediate risk (P = 0.004, odds ratio = 4.8, 95% CI 1.5 to 17.2), or both (P = 0.000006, odds ratio = 7.3, 95% CI 2.6 to 20.3). The difference between patients at low risk and those at intermediate risk was not statistically significant (P = 0.5, odds ratio = 0.4, 95% CI 0.02 to 2.6). After a median time of 36 months (range 9 to 119) from UUTT diagnosis, 5 (20%) of 25 patients have died of the disease. CONCLUSIONS: The incidence of metachronous UUTTs is low in patients with superficial bladder cancer at low or intermediate risk of disease recurrence or progression and significantly higher for patients at high risk. Because UUTT is often asymptomatic, and mortality is high, frequent and lifelong examination of the upper urinary tract is suggested, with an annual intravenous urogram and urinary cytologic analysis every 4 months in patients with superficial bladder cancer at high risk of disease recurrence or progression. PMID- 10367844 TI - Relation between urethral elasticity and bladder outlet obstruction and histologic composition of the prostate in patients with benign prostatic hyperplasia. AB - OBJECTIVES: In benign prostatic hyperplasia (BPH), bladder outlet obstruction is caused by mechanical blockage resulting from an enlarged prostate, and by the increased tone of prostatic smooth muscle. In patients with obstructive BPH, the prostatic urethra urodynamically corresponds to the flow-controlling zone (FCZ). We sought to investigate the relation between the obstruction and elastic properties of the FCZ and the histologic composition of the prostate in symptomatic patients with BPH. METHODS: The grade of infravesical obstruction was classified according to preoperative pressure-flow data from 30 men with symptomatic BPH, and the elastic properties of the FCZ were evaluated as the mean elastance, which was calculated using the three-parameter model. The area densities of smooth muscle, fibrous tissue, epithelium, and lumen were determined by quantitative morphometry, using BPH tissue obtained by transurethral resection. RESULTS: The area density of smooth muscle had a negative correlation with mean elastance (p = -0.50, P <0.01), the linear passive urethral resistance relation (p = -0.43, P <0.05), and the group-specific urethral resistance factor (p = -0.39, P <0.05). No other histologic element was correlated with these variables. CONCLUSIONS: Our results suggest that urethral elasticity and bladder outlet obstruction grade are influenced by the relative content of smooth muscle within the prostate in patients with BPH. PMID- 10367845 TI - Trends in prostatectomy for benign prostatic hyperplasia among black and white men in the United States: 1980 to 1994. AB - OBJECTIVES: To estimate the annual rate of discharge for prostatectomy for benign prostatic hyperplasia (BPH) in black and white men from 1980 to 1994 using the National Hospital Discharge Survey. METHODS: Overall and race-, age-, and year specific utilization rates were estimated for the civilian population in the United States. Length of stay was calculated for each discharge, and the results were plotted over time. An expected number of discharges based on the rates observed in 1980 was estimated to determine the impact of decreased prostatectomy rates on the number of procedures that would have been expected in this aging population. RESULTS: Discharge rates for whites were within a narrow range (233.2 to 274.5 per 100,000) from 1980 through 1990 and then displayed a monotonic decline after 1991 to 131.3 per 100,000 in 1994. Rates for blacks were 10% to 24% lower from 1980 to 1991; the decline in discharge rates began in 1993 for blacks, and by 1994 the racial gap had closed. Length of stay decreased throughout the period but length of stay averaged 30% longer for blacks throughout. On the basis of the observed rates of 1980, there were more than 140,000 fewer prostatectomies performed for BPH in 1994 than would have been expected owing to the aging of the population. CONCLUSIONS: These data demonstrate that the black/white differences in prostatectomy for BPH that were observed in the 1980s have disappeared in recent years. Furthermore, rates have declined dramatically in all age- and race specific groups. Further work is needed to determine whether this convergence in discharge rates is due to equalization of access to medical care or to differences in utilization of alternative therapies. PMID- 10367846 TI - Determination of alpha1-antichymotrypsin-PSA complex in serum does not improve the differentiation between benign prostatic hyperplasia and prostate cancer compared with total PSA and percent free PSA. AB - OBJECTIVES: To evaluate the analytical performance and diagnostic utility of alpha1-antichymotrypsin (ACT)-prostate-specific antigen (PSA) complex in serum to improve the differentiation between benign prostatic hyperplasia (BPH) and prostate cancer (PCa). METHODS: Serum concentrations of total PSA (tPSA), free PSA (fPSA), and ACT-PSA were measured in 112 untreated patients with PCa (median age 65 years), 34 patients with BPH (median age 66 years) with histologic confirmation, and 33 men without prostate disease and with a normal digital rectal examination considered as controls (median age 54 years). Sera were frozen at -80 degrees C within 2 hours after collection and then analyzed during a 12 week period. Determinations were made with the Enzymun-Test for tPSA and fPSA and with a prototype assay for ACT-PSA on the ES system (Roche Diagnostics, Boehringer Mannheim). RESULTS: The new ACT-PSA assay showed reliable data of analytical performance. The lower detection limit amounted to 0.068 microg/L. The assay was linear to 50 microg/L. Spiking experiments showed a mean recovery rate of 98.2%. No interferences of the assay were observed in patients with acute inflammation and highly increased ACT concentrations. The values of intra- and interassay imprecision ranged from 1.51% to 3.48% and 2.1% to 6.3%, respectively. The median value of ACT-PSA concentrations were significantly different (P <0.001) between controls and patients with BPH and PCa (0.40, 3.86, 5.26 microg/L, respectively). The median fPSA/tPSA and fPSA/ACT-PSA ratios were significantly different between BPH and PCa (24.3% versus 12.2%, P <0.001 and 32.9% versus 15.0%, P <0.001, respectively), but no difference of the ACT PSA/tPSA ratio was observed (78.2% versus 78.7%, P = 0.696). Receiver operating characteristics of ACT-PSA (area under the curve = 0.630) and all the derivative ratios of fPSA/ACT-PSA (area = 0.737) and ACT-PSA/tPSA (area = 0.528) were not different from that of tPSA (area = 0.619), but showed a lower discrimination power between BPH and PCa than the fPSA/tPSA ratio (area = 0.790). CONCLUSIONS: Using this prototype assay to quantify ACT-PSA in serum, we have demonstrated that ACT-PSA and the calculated derivatives are not superior in the differentiation between BPH and PCa compared with tPSA and the ratio of fPSA to tPSA. PMID- 10367847 TI - Impact of chronic dialysis on serum PSA, free PSA, and free/total PSA ratio: is prostate cancer detection compromised in patients receiving long-term dialysis? AB - OBJECTIVES: The increased incidence of malignancy (ie, prostate cancer) in patients with end-stage renal failure is well known. However, little is known of the impact of hemodialysis and various membrane types on total and free prostate specific antigen (PSA). We prospectively studied the impact of high- and low-flux dialysis membranes and kidney function on total PSA (tPSA), free PSA (fPSA), and free/total PSA ratio (f/t PSA). METHODS: A total of 149 men were included. tPSA, fPSA, and f/t PSA were measured before and immediately after dialysis with high flux (n = 101) and low-flux (n = 48) membranes in the serum and in the dialysis ultrafiltrate. A multivariate analysis of the impact of kidney function and age on the rate of change of all parameters was performed. RESULTS: Overall, a significant decrease of fPSA (from 0.49 +/- 0.3 to 0.35 +/- 0.3 ng/mL, P <0.0001) and f/t PSA (from 45 +/- 19% to 38 +/- 13%, P <0.0001) and a nonsignificant decrease in serum tPSA were observed. However, fPSA (from 0.51 +/- 0.5 to 0.27 +/ 0.3 ng/mL, P <0.0001) and f/t PSA (from 47 +/- 19% to 31 +/- 18%, P <0.0001) decreased significantly in high-flux membranes only. The ultrafiltrate contained 100% fPSA in high-flux membranes and no fPSA in low-flux membranes. Age, serum creatinine, blood urea nitrogen, and dialysis evaluation parameters (Kt/V) had no impact on correlation with changes in tPSA and fPSA. CONCLUSIONS: tPSA molecules do not pass high- and low-flux membranes; fPSA passes high-flux membranes only. The nonsignificant decrease of tPSA is due to adsorption to both dialysis membranes. Although tPSA can safely be used to screen patients on dialysis, independently from the dialysis procedure and membrane, fPSA and f/t PSA are only reliable with low-flux membranes. Finally, we can state that the fPSA is most probably cleared through the kidneys by glomerular filtration. PMID- 10367848 TI - Relationships between prostate-specific antigen and prostate volume in black and white men with benign prostate biopsies. AB - OBJECTIVES: To determine whether the higher age-adjusted serum prostate-specific antigen (PSA) levels in black compared with white men with no clinical evidence of prostate cancer reflect racial differences in relationships between PSA and prostate volume. METHODS: The age, PSA, findings on digital rectal examination (DRE), prostate volume, and PSA density were assessed prospectively in 810 consecutive, evaluable men who underwent prostate biopsy for suspected cancer but who had benign histologic findings. RESULTS: Among the black and white patients, there were significant differences in age (mean 67.2 +/- 8.1 and 65.9 +/- 7.7 years, respectively, P = 0.02), PSA (median 4.7 and 3.9 ng/mL, respectively, P <0.0001), prostate volume (median 41 and 36 mL, respectively, P = 0.004), and PSA density (median 0.11 and 0.08 ng/mL/mL, respectively, P = 0.005). Multiple linear regression analyses showed that black race was significantly associated with increased prostate volume when controlled for age (P = 0.02), with increased PSA when controlled for prostate volume and age (P = 0.002), and with increased PSA density when controlled for age (P = 0.007). When controlled for prostate volume, PSA was not significantly different in black and white men 50 to 59 years old but was significantly greater in black men 60 to 69 and 70 to 79 years old (P = 0.02 and 0.002, respectively). CONCLUSIONS: On a volume/volume basis, the benign prostatic tissue of black men appears to contribute more PSA to the circulating blood than does the benign prostatic tissue of white men, and the difference increases with advancing age. These phenomena provide a reasonable explanation for the age-adjusted racial differences in the PSA of men with no clinical evidence of cancer. PMID- 10367849 TI - Adenocarcinoma of the prostate in men younger than 40 years of age: diagnosis and treatment with emphasis on radical prostatectomy findings. AB - OBJECTIVES: Prostate cancer is rarely diagnosed in men younger than 40 years of age. It is thought, although not documented, that these tumors behave particularly aggressively. METHODS: We studied 87 men younger than 40 years old who underwent prostate needle biopsy and were from three populations: (a) 71 cases (63 benign, 7 cancer) from Dianon Systems; (b) 9 needle biopsies with cancer sent to one of us (J.I.E.) in consultation; and (c) 7 men with cancer who came to Johns Hopkins for consultation. RESULTS: The median age of men with a benign biopsy was 35 years (mean 33.9, range 22 to 39); the median age of men with cancer was 38 years (mean 35.9, range 22 to 39) (P = 0.004). The most common indications for biopsy were abnormal digital rectal examination (DRE) (n = 61); elevated prostate-specific antigen (PSA) (n = 14), and inflammatory symptoms (n = 12). Other reasons cited included hematuria, abnormal ultrasound, pain, ejaculatory problems, obstructive symptoms, and family history of prostate cancer. The median PSA was 2.6 ng/mL (mean 4.8, range 0.3 to 66) for all men, 1.2 ng/mL (mean 3.4, range 0.3 to 19.9) for benign cases, and 4.4 ng/mL (mean 8.7, range 2.1 to 66) for cancer (P = 0.0004). Abnormal DRE was not predictive of cancer. Of the 55 patients whose family history was known, 40 men had no family history of prostate cancer, and of those, only 6 (15%) had cancer. Of the 1 5 patients with a family history of cancer, 6 (40%) were found to have cancer on biopsy (P = 0.05). Of the 23 patients with cancer, 3 were lost to follow-up, 1 was treated with hormones, and 3 chose watchful waiting. The remaining 16 patients underwent radical prostatectomy and had diverse pathologic findings. Tumor volume ranged from 0.01 to 6.35 cc. Pathologic stage was pT2 in 9 cases and pT3 in 7 cases (2 with positive pelvic lymph nodes). In 14 men, serum PSA values were available: of 4 men with PSA greater than 10 ng/mL, all had Stage pT3, and of 10 men with PSA less than 10 ng/mL, 3 had Stage pT3. CONCLUSIONS: Young men who are candidates for radical prostatectomy have potentially curable disease, particularly if PSA at the time of diagnosis is less than 10 ng/mL. PMID- 10367850 TI - Adjuvant radiotherapy in patients with pathologic Stage C (pT3N0) adenocarcinoma of the prostate. AB - OBJECTIVES: This report is an update on the outcomes in the management of pathologic Stage C (T3N0) prostate cancer (CaP) with postoperative irradiation. METHODS: Between 1976 and 1994, 311 patients with pathologic Stage C CaP were treated with radical prostatectomy. Pathologic stage was as follows: C1, 60 patients (19%), C2, 146 patients (47%), and C3, 105 patients (34%). Gleason score was 2 to 4 in 10 patients (3.2%), 5 to 6 in 121 (39%), 7 in 101 (32%), and 8 to 10 in 76 (24%); median prostate-specific antigen (PSA) level was 11.9 ng/mL. Postoperative irradiation consisted of a median dose of 48 Gy. Follow-up was up to 18 years (median 5). RESULTS: The 10-year actuarial survival was 81% and 10 year disease-free survival was 51%. Pathologic stage and Gleason score were independently predictive of recurrence, each with P >0.001 after controlling for the other. Patients with pathologic Stage C3 and Gleason score 7 to 10 were in the worst prognostic category and had 5.4 times the risk of recurrence compared with patients with pathologic Stage C1-C2, Gleason score 2 to 6. Preoperative PSA was a good (P = 0.02) predictor of disease-free survival. Clinical recurrence was seen in 28 patients (9%), including 10 (3.2%) with local recurrence. PSA recurrence (PSA greater than 0.05 ng/mL) developed in 68 patients (22%). CONCLUSIONS: With the known limitations of a nonrandomized clinical trial, on the basis of the experience of this study we recommend the use of moderate dose, limited-field postoperative radiotherapy in patients with pathologic Stage C disease with Gleason score greater than 4. PMID- 10367851 TI - Prostate cancer: demographic and behavioral correlates of stage at diagnosis among blacks and whites in North Carolina. AB - OBJECTIVES: Although stage at diagnosis is one of the most important predictors of survival from prostate cancer, demographic factors, screening practices, and knowledge and beliefs associated with stage at diagnosis have not been well documented, particularly by race. METHODS: We conducted telephone interviews with 117 black and 114 white men diagnosed with prostate cancer to identify the demographic factors, healthcare-seeking behaviors, and prostate cancer-related knowledge, attitudes, and practices associated with stage. The sample was stratified by stage at diagnosis and was composed of men 50 to 74 years old who resided in a contiguous 63-county region in North Carolina and who were diagnosed at 1 of 16 participating hospitals. RESULTS: Among blacks, stage was inversely correlated with income (P = 0.04) and health insurance status (P < or = 0.001); among whites, stage was not associated with income or health insurance status, but approached significance with marital status (P = 0.06). Awareness of prostate cancer before diagnosis tended to decline with advancing stage among black men (P = 0.07), but was high for all stages (greater than 93%) among whites. Report of a prostate-specific antigen screen was inversely correlated with stage among black men (P = 0.01); a trend was observed among whites but was not significant (P = 0.20). Knowledge of prostate cancer risk factors was not significantly associated with stage for blacks or whites. Less than one third of men in each race and stage group knew that black men are at increased risk of prostate cancer. CONCLUSIONS: Demographic and other factors vary with stage and should be considered when designing and targeting interventions to reduce late diagnosis of prostate cancer. PMID- 10367852 TI - Nocturnal electrobioimpedance volumetric assessment of patients with erectile dysfunction. AB - OBJECTIVES: Electrobioimpedance volumetric assessment is a procedure that can measure penile length, cross-sectional area, and volume. From these variables, the number and duration of erectile events, volume change, and percentage of volume increase from baseline can be determined. This procedure was performed on patients with erectile dysfunction (ED) and findings were compared with patients with no history of ED. Examples of etiology are reported. METHODS: Two groups of patients with ED were evaluated by electrobioimpedance assessment. Group 1 patients (n = 23), ranging in age from 26 to 60 years (mean 50), were involved in simultaneous electrobioimpedance assessment and duplex Doppler ultrasound penile volume measurements. A tissue correction was derived. Group 2 patients (n = 10), ranging in age from 38 to 64 years (mean 50), used nocturnal electrobioimpedance volumetric assessment (NEVA) at home for 2 consecutive nights. RESULTS: After deriving an expression to correct for tissue volume, simultaneous measurement of penile blood volume by NEVA and duplex Doppler showed that the regression line for study participants and the identity line was not significantly different by analysis of variance. Using NEVA in comparing patients with ED to a reference population with no history of ED, and using a two-tailed Student's t test for means, the data demonstrated a statistically significant (P < or =0.05) difference in the number of erectile events and percentage of volume change over baseline. With NEVA data, it was possible to distinguish arterial insufficiency from veno-occlusive dysfunction. CONCLUSIONS: The present study demonstrates that electrobioimpedance volumetric assessment can be used in patients with ED. Compared with a reference population with no history of ED, the group with ED had fewer nocturnal erectile events that resulted in a smaller increase in penile blood volume change over baseline. Although the time dependence of the measured variables identifies the cause of ED, the application of NEVA to a larger population will allow further analysis of the dynamic information contained in the NEVA data. PMID- 10367853 TI - Traumatic posterior urethral injury and early primary endoscopic realignment: evaluation of long-term follow-up. AB - OBJECTIVES: The management of complete or partial posterior urethral disruption is controversial and much debate continues regarding immediate versus delayed definitive therapy. We further analyze our experience and long-term results using early endoscopic realignment. METHODS: Between April 1991 and June 1995, 8 men with posterior urethral avulsion, either complete or partial and secondary to blunt trauma and pelvic fractures, presented to our institution. A variety of endourologic techniques were employed to achieve urethral continuity while attempting to minimize stricture formation, incontinence, and impotence. RESULTS: After a mean of 50.4 months (range 35 to 85) of follow-up, 7 men (87.5%) are continent, with 2 of those requiring intermittent self-dilation ranging from once every 7 days to once a month. One patient required conversion to an open perineal urethroplasty. Of the 8 patients, 5 (62.5%) are potent, and 2 others achieve adequate erections for intercourse using intracorporeal injections. Four of the 8 have required subsequent internal urethrotomies with eventual voiding stabilization over the course of 1 2 months. Average time to realignment was 9.5 days (range 0 to 19). CONCLUSIONS: Primary endoscopic realignment offers an effective method for treating traumatic urethral injuries. Our long-term follow up provides further support for use of this technique by demonstrating that urethral continuity can be established without increased incidence of impotence, stricture formation, or incontinence. By achieving early and minimally invasive realignment, we seem to lessen the severity of stricture disease that almost uniformly afflicts those patients who undergo delayed repair. If a minimally invasive technique should fail, it does not seem to delay nor does it preclude further management using open techniques. PMID- 10367854 TI - Color flow Doppler sonography: a reliable alternative to voiding cystourethrogram in the diagnosis of vesicoureteral reflux in children. AB - OBJECTIVES: In children with urinary tract infection, the incidence of vesicoureteral reflux (VUR) is nearly 30% to 40%. The standard for the diagnosis of VUR is voiding cystourethrography (VCUG). This study assessed the role of color flow Doppler sonography (CFDS) in the diagnosis of VUR and ureteral jets. METHODS: CFDS imaging was performed in 36 patients aged 6 months to 13 years during a 4-year period. All patients underwent CFDS and VCUG within 24 to 48 hours, but the findings of the VCUG were not reported to the sonologist. The ultrasound examinations were done using a color Doppler real-time machine. Representative images of the bladder events were recorded with a multiformat camera and on VHS videotape. RESULTS: The duration of the Doppler signal varied from 0.4 to 7.5 seconds. In 31 (86.1%) of 36 patients, the results of CFDS correlated well with VCUG findings. There were three false-negative and two false positive results in the present study. Six patients underwent reimplantation during the course of their treatment. CFDS was used as a follow-up modality at the end of 6 months, and the results correlated well with standard VCUG in 4 of these patients. In the remaining 2 patients, only CFDS was performed and correlation with VCUG was not possible. VCUG was considered the reference standard in assessing the sensitivity of CFDS. CONCLUSIONS: CFDS of the bladder during the filling and micturating phases is a reliable and sensitive modality for identifying VUR and demonstrating ureteral jets. CFDS nullifies the danger of exposure to ionizing radiation and avoids the unpleasant catheterization many of these children fear. PMID- 10367855 TI - Evaluation of the functional significance of the colorectal valve used in rectal urinary diversion in children: a comparative study between cases with and without the valve. AB - OBJECTIVES: To study the long-term impact of functional isolation of rectal reservoirs on the blood chemistries and acid-base balance of children who underwent this type of urinary diversion. METHODS: A retrospective evaluation of 63 children with rectal reservoirs was performed. Of these, 40 had a colorectal valve and 23 had double-folded rectal reservoirs without a functional isolation valve. Evaluation included serum chemistry and arterial blood sample analysis to verify the impact of the created valve on homeostasis. RESULTS: There was a statistically significant difference between the two groups relative to the value of pH, P(CO2), bicarbonate, base excess, and chloride in favor of those having a colorectal valve. Reduction of the absorptive surface area of the colon is presumably the cause in view of the functional isolation created by the colorectal valve. CONCLUSIONS: In children, the long life expectancy and the benign condition for which they have diversion require the incorporation of the colorectal valve in any form of continent rectal diversion. Prophylactic alkalinization with strict follow-up is a must in those having no valve. PMID- 10367856 TI - Removal of retained Penrose drain under fluoroscopic guidance. AB - An uncommon complication of Penrose drain usage is retention of the drain by a fascial suture. Removal of a retained Penrose drain can be carried out percutaneously under fluoroscopic guidance. PMID- 10367857 TI - Initial experience with processed human cadaveric allograft skin for reconstruction of the corpus cavernosum in repair of distal extrusion of a penile prosthesis. AB - We describe our initial experience with the novel application of processed human cadaveric allograft skin in reconstruction of a damaged corpus cavernosum associated with distal extrusion of a penile prosthesis. The material was evaluated for ease of reconstruction, adequacy of repair, and outcome. Human processed dermis allograft requires no intraoperative harvesting, is technically easy to fashion, and offers adequate tensile strength in the reconstruction of damaged corpora cavernosa. This initial experience with processed human cadaveric dermis in reconstruction of damaged corpora cavernosa is encouraging. Further evaluation to define the long-term efficacy and scope of application of this material in urologic reconstructive procedures is warranted. PMID- 10367858 TI - Images in clinical urology. Giant angiomyolipoma. PMID- 10367859 TI - Unilateral renal cystic disease. PMID- 10367860 TI - Possible causes for the low prevalence of pediatric urolithiasis. AB - OBJECTIVES: To determine why the incidence of pediatric urolithiasis is less than that of adult urolithiasis, we investigated the difference in inhibition of calcium oxalate (CaOX) crystallization between pediatric and adult urinary macromolecules (UMMs). METHODS: Urinary parameters in relation to urolithiasis, the inhibition of CaOX crystallization of original urine and urine from which UMMs (greater than 3 kDa) had been removed, and the inhibition of CaOX crystal growth and aggregation of UMMs alone were measured. These inhibitory activities were compared between children and adults. RESULTS: In the original urine, the inhibition of CaOX crystallization was significantly stronger for children than for adults, but was the same in urine from which the UMMs had been removed. The inhibition of CaOX crystal growth by UMMs alone showed no significant differences between children and adults; their inhibition of CaOX crystal aggregation was significantly stronger for children than for adults. Much more glycosaminoglycan (GAG) was included in pediatric UMMs than in adult UMMs, although there was no difference in UMM concentration between urine from children and urine from adults. CONCLUSIONS: The lower incidence of CaOX lithiasis in children may be attributed, among other factors, to the stronger inhibition of CaOX crystal aggregation by pediatric UMMs, which in turn might be affected by the higher concentration of GAGs in children's urine. PMID- 10367862 TI - Comparison of the breaking strength of polyglactin mesh in urine, serum, and cell culture media. AB - OBJECTIVES: This study was designed to determine the durability of polyglactin woven mesh in various in vitro environments, including urine, since polyglactin 901 mesh has been considered for implementation in urinary tract reconstruction. METHODS: Segments of 1 x 1-cm sterile woven and knitted polyglactin 910 mesh with and without collagen coating were exposed to the following conditions: dry (at room temperature and at 37 degrees C, humidified air), in porcine and human serum and urine, in porcine urine over a range of pH levels, in infected urine, in cell culture media (MCDB 105 with 5% fetal bovine serum), and in cell culture media with porcine bladder fibroblasts. The mesh breaking strength was measured at 0, 12, 21, 28, and 36 days. RESULTS: The mean breaking strength for dry, room temperature mesh segments measured 350 g for all time intervals. At day 21, the breaking strength for all mesh types in human and porcine serum, cell culture media, and cell culture media with bladder fibroblasts was less than 10% of the control, but the human and porcine urine maintained 12% to 24% of the control breaking strength (this difference did not reach statistical significance). There was no significant difference in the breaking strength in human and porcine urine or human and porcine serum. By day 38, the breaking strength for all mesh types in all solutions was less than 5% of the control breaking strength. The presence of fibroblasts increased the rate of degradation of the mesh compared with the urine, serum, and cell culture media alone. There was a significant prolongation of degradation with decreasing pH, as well as with infected urine. This prolongation was additive; in fact, all mesh types in low pH (5.0), infected urine showed minimal degradation at 38 days. CONCLUSIONS: In acidic infected urine, the durability of polyglactin 910 mesh is significantly prolonged compared with the other conditions tested. Therefore, when used in urinary tract reconstruction, as in other organ systems, the integrity of the polyglactin mesh should diminish rapidly after 3 weeks as long as the urine is kept sterile and a neutral to alkaline pH is maintained. PMID- 10367861 TI - Comparison of absorbable and nonabsorbable sutures for microsurgical vasovasostomy in rats. AB - OBJECTIVES: At least 12% of initially patent vasovasostomies (VVs) shut down. Currently, only nonabsorbable sutures are used for VV. A synthetic, slowly absorbing, monofilament polyglactin suture has been developed that retains tensile strength for up to 6 months. We performed a prospective controlled randomized study comparing absorbable and nonabsorbable sutures for rat VVs. METHODS: Bilateral microsurgical VV was performed in three groups of 36 Wistar male rats, with 10-0 nylon, 10-0 polypropylene, and 10-0 polyglactin sutures. Twelve control rats underwent sham operations. Three rats in each group were killed at 2, 6, 12, and 24 weeks. The abdominal end of the vas deferens was transected and the intraluminal fluid examined microscopically for presence of sperm. The segment of the vas deferens containing the anastomosis was excised. Fluid from the testicular end was examined for sperm to confirm spermatogenesis. Patency was confirmed by an antegrade indigo carmine vasogram of the anastomotic segment. Segments were randomly sent for histologic or tensile strength evaluation. RESULTS: The mean tensile strength of the anastomoses performed with nylon was slightly higher than in polypropylene and polyglactin sutures, although the difference was not statistically significant. Polyglactin consistently maintained tensile strength throughout 6 months without significant fluctuations. The mean patency rate in the polyglactin group was 96%, in nylon 81%, and in polypropylene 61%. Although polyglactin had a consistently higher patency rate compared with nonabsorbable sutures, the difference was not statistically significant (P = 0.11) but indicated a strong trend. The occurrence of microscopic sperm granuloma, muscle layer injuries, intimal fibrosis, and adventitial fibrosis of the vas deferens was not significantly different between suture types. CONCLUSIONS: The three suture materials appear equivalent with respect to overall tensile strength of anastomosis; with histologic evaluation, the trend was toward better patency with polyglactin. Polyglactin 10-0 microsurgical suture is a viable alternative to nonabsorbable sutures in microsurgical VVs, although further studies are indicated to assess long-term results. PMID- 10367863 TI - Transrectal ultrasound-guided intraprostatic injection of absolute ethanol with and without carmustine: a feasibility study in the canine model. AB - OBJECTIVES: To develop a reliable intraprostatic injection technique and to define the local and systemic toxicity of intraprostatic injection of dehydrated ethanol with and without carmustine. METHODS: Twenty-three random-source male canines were divided into a control group (n = 3), a dehydrated ethanol-alone group (group 1, n = 10), and a dehydrated ethanol-plus-carmustine group (group 2, n = 10). A reliable intraprostatic injection technique was developed with the control animals. The optimal volume of dehydrated ethanol for intraprostatic injection and the local tissue effects of dehydrated ethanol injection were defined with group 1. The local tissue effects of escalating doses of carmustine were defined with group 2. All animals were injected under general anesthesia using transrectal ultrasound (TRUS) guidance. Fourteen days after injection, a repeated TRUS of the prostate was done, the animals were killed, and the bladder, prostate, and periprostatic tissues were excised for pathologic examination. RESULTS: Sonographic changes in the prostate 2 weeks after injection were present in all group 1 and 2 animals. All prostates had varying amounts of hemorrhagic and coagulative necrosis, which correlated with the TRUS findings. There were no differentiating pathologic features between group 1 and group 2 specimens. The relative amount of necrosis varied with the doses of dehydrated ethanol and carmustine injected, but was not predictable on the basis of the doses administered. Subclinical prostatic microabscesses were identified in 6 of 10 group 1 animals and 4 of 10 group 2 animals. Only group 2 animals had alterations in their blood chemistry results, all of which were self-limited. Two had white blood cell nadirs of less than 2000 5 days after injection. No animals developed incontinence, and there were no rectal injuries. CONCLUSIONS: Intraprostatic dehydrated ethanol and carmustine injections were readily controllable under TRUS guidance and resulted in hemorrhagic and coagulative necrosis of prostatic tissue with minimal associated morbidity and no incontinence in the dog model. Hematologic changes observed in the animals that received carmustine were self limiting. PMID- 10367865 TI - Effect of surgically induced varicocele on testicular blood flow and Sertoli cell function. AB - OBJECTIVES: To evaluate the effect of varicocele on testicular blood flow and expression by Sertoli cells of transferrin and androgen-binding protein (ABP), to determine whether varicoceles impair Sertoli cell function. METHODS: Experimental varicocele was established in male Sprague-Dawley rats by partial ligation of the left renal vein. The control group received a sham operation. At 30 minutes after surgery, rats underwent a xenon-133 washout study, and at 30 days after surgery, transferrin, ABP, and testicular blood flow were evaluated. Expression of transferrin and ABP were evaluated using immunohistochemical techniques. Testicular blood flow was measured using xenon-133 clearance techniques. Statistical analyses were done with an independent t test. RESULTS: The testicular blood flow was 16.7 +/- 1.25 mL/100 g/min in varicocele-bearing rats and 21.01 +/- 0.46 mL/100 g/min in sham-operated rats 30 minutes after surgery. Testicular blood flow remained decreased at 30 days in varicocele-bearing rats (15.12 +/- 1.08 mL/100 g/min) and remained stable in the control group (19.45 +/- 0.55 mL/100 g/min). The expression of transferrin and ABP was significantly reduced in varicocele-bearing rats compared with the control group. CONCLUSIONS: Our study suggests that a decrease in testicular blood flow may lead to impaired Sertoli cell function in varicocele-bearing rats. PMID- 10367864 TI - Endogenously formed nitric oxide modulates cell growth in bladder cancer cell lines. AB - OBJECTIVES: Nitric oxide (NO) is formed in many mammalian tissues, and a growing body of evidence suggests that NO is involved in cell growth and cell differentiation. Low concentrations of NO can stimulate cell growth; high concentrations result in cytostatic/cytotoxic effects. It has previously been shown that intravesical treatment with bacille Calmette-Guerin (BCG) for bladder cancer increases NO production in the human urinary bladder and that NO inhibits bladder cancer cell growth in vitro. In this study, we investigated nitric oxide synthase (NOS) activity in different bladder cancer cells and the role of the NO precursor L-arginine in cell proliferation. METHODS: NOS activity was assessed by citrulline assay in cultured normal human urothelial cells and bladder cancer cell lines T24 and MBT-2 before and after treatment with cytokines. We also measured cell growth at various L-arginine concentrations and after addition of the NOS inhibitor N(G)-nitro-L-arginine (L-NNA) in unstimulated and cytokine stimulated cells. RESULTS: Normal urothelial cells, as well as T24 and MBT-2 cells, showed calcium-dependent NOS activity under basal conditions. The bladder cancer cell lines also showed calcium-independent NOS activity in contrast to the normal cells. After cytokine treatment, both the normal cells and the cancer cell lines showed a marked increase in calcium-independent NOS activity. There was a dose-dependent stimulation of cell growth in the cancer cell lines after L arginine addition, and this effect could be antagonized by L-NNA. Cytokine treatment inhibited cell growth, and this inhibition was partly reversed by L NNA. CONCLUSIONS: Normal urothelial cells and bladder cancer cell lines MBT-2 and T24 show NOS activity, and cytokine treatment induces calcium-independent NOS activity. Our results suggest that endogenous activity of the constitutively expressed form of NOS in unstimulated cells promotes cell proliferation, and NO production secondary to increased activity of the inducible form of NOS after cytokine treatment inhibits cell growth. PMID- 10367866 TI - Presidential Address. Visions: medical education and surgical training in evolution. PMID- 10367867 TI - Adenosquamous carcinoma of the pancreas. AB - HYPOTHESIS: Adenosquamous carcinoma of the pancreas is a rare but particularly virulent variant of invasive ductal carcinoma. This review will demonstrate the aggressive biologic activity, histopathologic features, and DNA flow cytometric characteristics of this aggressive lesion. In addition, the outcome is less favorable than in other pancreatic neoplasms, in spite of aggressive surgical and postoperative adjuvant therapy. DESIGN: A retrospective review of 6 patients treated during an 8-year period. SETTING: A major urban university tertiary referral hospital. PATIENTS: There were 6 patients with this unusual tumor seen between 1990 and 1998. There were 4 men and 2 women, all white, with a mean+/-SD age of 63.5+/-14.7 years. Symptoms were similar to those in patients with more common pancreatic malignant neoplasms. RESULTS: Four patients with tumors in the head of the pancreas had pancreatoduodenectomy, and 2 with body and or tail lesions had distal pancreatectomy and splenectomy. Pathologically, all the tumors were poorly differentiated and aneuploid, and 5 of the 6 were locally metastatic. All but 1 patient had postoperative complications, but there were no operative deaths. One half of the patients received postoperative adjuvant chemotherapy and radiation therapy. Only 1 patient is still alive at 9 months after surgery, but has known residual cancer around his portal vein noted during palliative distal pancreatectomy. CONCLUSIONS: Adenosquamous carcinoma of the pancreas is an uncommon variant of exocrine pancreatic neoplasm. It is characterized by an admixture of adenomatous and squamous cell elements and demonstrates aggressive biologic behavior. This series of 6 patients is similar to the 134 cases reported since 1907, in that survival is short despite aggressive surgical therapy. Few patients with this disease live more than 1 year. Aggressive therapy should be tempered by the realization of the uniform poor prognosis associated with this malignant neoplasm. PMID- 10367868 TI - Long-term results of biliary reconstruction after laparoscopic bile duct injuries. AB - HYPOTHESIS: The Hepp-Couinaud approach to biliary enteric reconstruction for laparoscopic bile duct injuries provides a durable, long-term result in most patients. DESIGN: Retrospective study of patients who underwent operative repair of laparoscopic bile duct injuries from January 1990 through December 1997. SETTING: Academic tertiary referral center. MAIN OUTCOME MEASURES: Outcome was assessed using a grading system based on clinical symptoms, liver function tests, and need for reintervention for anastomotic stricture. The Kaplan-Meier method was employed to estimate stricture-free survival. RESULTS: Fifty-nine consecutive patients underwent operative repair of the following laparoscopic bile duct injuries (Strasberg classification): B: n = 2 (3%), C: n = 1 (1%), D: n= 2 (3%), E1: n= 5 (8%), E2: n= 16 (27%), E3: n= 25 (42%), E4: n = 5 (8%), and E5: n = 3 (5%). Forty-seven patients (80%) had 1 or more interventions prior to the index repair. The extrahepatic left bile duct (Hepp-Couinaud approach) was used in 46 of 53 patients who underwent a Roux-en-Y hepaticojejunostomy. Follow-up (mean+/ SEM, 3.7+/-0.3 years) was complete in 54 of the 57 patients still alive. Five patients developed subsequent anastomotic strictures and were treated with percutaneous transhepatic dilation (n = 3), endoscopic dilation (n = 1), and operative revision (n= 1). Excellent to good long-term results were achieved in the remaining 49 patients (91%). Life-table analysis yielded 95% and 88% chances of stricture-free survival at 2 and 5 years, respectively. CONCLUSIONS: Complex iatrogenic proximal bile duct injuries and strictures are amenable to operative repair using the extrahepatic left bile duct. The Hepp-Couinaud approach offers a durable result in more than 90% of patients, even after previous interventions have failed. PMID- 10367869 TI - Prophylactic antibiotics for elective laparoscopic cholecystectomy: are they necessary? AB - HYPOTHESIS: Prophylactic antibiotic treatment in elective laparoscopic cholecystectomy does not lower the already low infection rate associated with this procedure. DESIGN AND SETTING: Prospective double-blind randomized trial at a community-based training hospital. PATIENTS: Four hundred fifty patients undergoing elective laparoscopic cholecystectomy were randomized into 1 of 3 treatment arms: (1) preoperative cefotetan disodium, 1g intravenously; (2) preoperative cefazolin, 1g intravenously; and (3) intravenous placebo. There were no demographic differences between groups in age, smoking history, American Society of Anesthesiologists score, infection risk class, time of antibiotic administration prior to surgery, and type of skin preparation. INTERVENTIONS: Laparoscopic cholecystectomy was attempted in all cases; however, 10 patients required conversion to an open cholecystectomy and they were included in the statistical analysis. Preoperatively, all patients were randomized in a blinded manner and received cefotetan, cefazolin, or placebo intravenously. RESULTS: There were 10 postoperative infections. In the cefotetan group, there were 3 cases of superficial surgical site infections. In the cefazolin group, there were 2 superficial surgical site infections-1 pneumonia and 1 rhinosinusitis. In the placebo group, there were 2 superficial surgical site infections and 1 urinary tract infection. The overall infection rate in this series was 2.4%. Follow-up was performed at routine postoperative visits and by telephone contact. Data were evaluated using the chi2 test and analysis of variance with Duncan post hoc test (P<.05). CONCLUSION: Based on our data, use of prophylactic antibiotics does not decrease the rate of wound infections in elective laparoscopic cholecystectomy. PMID- 10367870 TI - Complex aortofemoral prosthetic infections: the role of autogenous superficial femoropopliteal vein reconstruction. AB - BACKGROUND: With increasing experience, we have encountered patients with complex aortofemoral prosthetic infections in whom extra-anatomic bypass (EAB) is not an option. HYPOTHESIS: Autogenous superficial femoropopliteal vein (SFPV) aortic reconstruction provides a limb-saving and lifesaving alternative with acceptable morbidity and mortality. DESIGN: Retrospective review. SETTING: University-based county, private, and Veterans Affairs hospitals. PATIENTS: Seventeen patients with infected aortofemoral bypasses in whom conventional EAB was impossible because of infection of previously placed EAB, massive groin and/or thigh sepsis, or both. MAIN OUTCOME MEASURES: Morbidity and mortality. RESULTS: Multiple previous operations were common (mean, 4 per patient) and included EAB (n = 11), replacement aortofemoral bypass (n = 4), prosthetic femoropopliteal bypass (n = 7), and thoracobifemoral bypass (n = 1); all bypasses became infected. Overall, 11 patients had sepsis at the time of presentation. Of the patients with massive groin infection, 7 had extensive deep infections involving most of the proximal thighs or retroperitoneum, 4 had enterocutaneous fistulae, and 2 had necrotizing fasciitis of the lower abdomen and thigh. Polymicrobial infections were common (n = 9). Four patients (24%) died in the perioperative period, 8 (47%) suffered major complications, and 4 (24%) underwent major amputations. Mortality in this group of patients was 3 times that of all other patients undergoing autogenous SFPV aortic reconstruction for prosthetic infection (8%). Amputation rates were also increased (24% vs 6%). The mean+/-SD follow-up time is 23+/-21 months. All patients maintained patent SFPV reconstructions. CONCLUSIONS: In the setting of complex aortofemoral prosthetic infections, autogenous SFPV aortic reconstruction is a useful option for patients in whom EAB is impossible and limb loss and/or death would be inevitable without revascularization. PMID- 10367871 TI - Oral contrast solution and computed tomography for blunt abdominal trauma: a randomized study. AB - HYPOTHESIS: Oral contrast solution (OC) is unnecessary in the acute computed tomographic (CT) evaluation of the patient with blunt abdominal trauma. DESIGN: Randomized controlled clinical trial. SETTING: Level I trauma center at a university-affiliated teaching hospital. PATIENTS: Five hundred adult patients sustaining blunt abdominal trauma and requiring urgent resuscitation and CT evaluation of the abdomen were eligible for the study. Those patients who were younger than 18 years, pregnant, or in police custody were excluded. One hundred six patients were excluded from the analysis (15 for inappropriate enrollment, 9 because a CT scan had not been performed, 1 owing to inability to accept a nasogastric tube, and 81 owing to missing or incomplete records). Three hundred ninety-four patients with an average age of 36 years, an average Revised Trauma Score of 10, and an average Glasgow Coma Scale score of 12 are included in the analysis. INTERVENTIONS: Patients were randomized via computer-generated assignment to 1 of 2 groups either receiving OC or not receiving OC (no OC) after placement of a nasogastric tube. All patients received intravenous contrast solution and then underwent helical CT scan of the abdomen and pelvis using the GE HiSpeed Advantage CT scanner (GE Medical Systems, Milwaukee, Wis). MAIN OUTCOME MEASURES: Abnormal CT results, need for laparotomy, missed gastrointestinal tract and solid organ injuries, nausea, and vomiting. RESULTS: There were 199 patients in the OC group and 195 patients in the no OC group. Vomiting occurred in 12.9% of patients and the incidence was not different between groups. One hundred five abnormal scans (50 OC and 55 no OC) were obtained and 33 patients with abnormal scans (19 OC and 14 no OC) underwent laparotomy. There was 1 nontherapeutic laparotomy in each group. There was 1 missed small-bowel injury in the OC group (sensitivity, 86%) and no missed small bowel injuries in the no OC group (sensitivity, 100%). Six bowel injuries were identified at laparotomy in the OC group. Two of the injuries were perforations without contrast extravasation but with pneumoperitoneum in 1. Three bowel injuries were identified in the no OC group, none of which were perforations. Seven of the 9 patients with bowel injury at laparotomy had associated intra abdominal injury. Specificity for solid organ injury was 94% in the OC group and 57.1% in the no OC group. Sensitivity for solid organ injury was 84.2% in the OC group and 88.9% in the no OC group. The average time to abdominal CT scanning after placement of a nasogastric tube was 39.02+/-18.73 minutes in the no OC group and 45.92+/-24.17 minutes in the OC group (P= .008). CONCLUSION: The addition of OC to the acute CT protocol for the evaluation of the patient with blunt abdominal trauma is unnecessary and delays time to CT scanning. PMID- 10367872 TI - Laparoscopic vs open adrenalectomy for the treatment of primary hyperaldosteronism. AB - HYPOTHESIS: That the clinical presentations, biochemical profiles, and surgical outcomes of patients treated with laparoscopic vs open adrenalectomy for primary hyperaldosteronism are different. DESIGN, SETTINGS, PATIENTS, AND INTERVENTIONS: The medical records of 80 patients with primary hyperaldosteronism who underwent open adrenalectomy between 1975 and 1986 or laparoscopic adrenalectomy between 1993 and 1998 at the University of California-San Francisco were reviewed by a single unblinded researcher (W.T.S.). MAIN OUTCOME MEASURES: Severity of hypertension and hypokalemia at diagnosis, their improvement after adrenalectomy, and operative complications. RESULTS: Thirty-eight patients underwent open adrenalectomy and 42 patients underwent laparoscopic adrenalectomy. The patients who underwent open adrenalectomy had documented hypertension for a median of 5 years before surgery; all had diastolic blood pressures greater than 100 mm Hg. Laparoscopically treated patients had documented hypertension for a median of 2.5 years preoperatively, and 20 (48%) had diastolic blood pressures greater than 100 mm Hg. The median preoperative serum potassium levels for the open and laparoscopic groups were 2.6 mmol/L and 3.3 mmol/L, respectively; the mean serum aldosterone levels were 1.47 nmol/L and 1.30 nmol/L. Thirty-two (84%) of the 38 patients who underwent open surgery and 41 (98%) of the 42 patients treated laparoscopically had adrenal adenomas. The sensitivity of preoperative computed tomographic scanning for adenomas was 83% for the patients treated with open adrenalectomy and 93% for those treated laparoscopically. There were 4 postoperative complications in the open surgery group and none in the laparoscopic group. Postoperatively, 30(81%) of 37 patients (excluding 1 patient who died of adrenocortical carcinoma) in the open surgery group and 37 (88%) of 42 patients treated laparoscopically were normotensive. Post-operative values were 3.6 to 5.0 of serum potassium per liter and 3.5 to 4.9 of serum potassium per liter in the open and laparoscopic groups, respectively. CONCLUSIONS: Patients who are treated with laparoscopic adrenalectomy for primary hyperaldosteronism are being referred with less severe hypertension and hypokalemia than patients formerly treated with open adrenalectomy. Patients treated laparoscopically had fewer postoperative complications and were equally likely to improve in blood pressure and hypokalemia. Laparoscopic adrenalectomy has become the treatment of choice for patients with primary hyperaldosteronism because of lower morbidity. PMID- 10367873 TI - Short esophagus: analysis of predictors and clinical implications. AB - HYPOTHESIS: Preoperative assessment can identify the predictors of esophageal shortening in patients with gastroesophageal reflux disease. DESIGN AND SETTING: Patient comparison study in a university-based tertiary care center. PATIENTS: A total of 236 patients with gastroesophageal reflux disease underwent primary antireflux procedures. Sixty-five patients were suspected of having a short esophagus and underwent a transthoracic approach. In 37 patients, a lengthening procedure was necessary to avoid tension on the repair. The remaining 28 patients were thought-after complete esophageal mobilization-to have sufficient length for a repair without needing a gastroplasty. An abdominal approach (laparoscopic Nissen fundoplication) was performed on 171 patients judged to have normal esophageal length. MAIN OUTCOME MEASURES: Univariate and multivariate analyses of preoperative variables were performed to identify predictors of a short esophagus. RESULTS: On univariate analysis, manometric esophageal length below the fifth percentile of normal was associated with esophageal shortening. On multivariate analysis, only the presence of an esophageal stricture predicted the need for a Collis gastroplasty (odds ratio, 7.5). The presence of Barrett's esophagus of 3 cm or greater identified patients in whom the transthoracic esophageal mobilization alone was sufficient (odds ratio, 3.4). CONCLUSIONS: The presence of a stricture was associated with esophageal shortening sufficient to require a gastroplasty. Transthoracic esophageal mobilization alone was usually sufficient to perform a safe repair without tension in patients with a Barrett's esophagus of 3 cm or greater. PMID- 10367874 TI - Endoscopic ultrasound and fine needle aspiration for the evaluation of pancreatic masses. AB - HYPOTHESIS: Endoscopic ultrasound (EUS) and endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) are accurate for the preoperative staging of pancreatic ductal carcinoma. DESIGN: Retrospective medical record review. PATIENTS: A prospective registry of 98 patients having EUS-FNA for peripancreatic masses from April 1994 to April 1998 was analyzed. MAIN OUTCOME MEASURE: The accuracy of EUS-FNA for preoperative diagnosis and staging of peripancreatic neoplasms. RESULTS: Ninety-eight patients, aged 41 to 91 years (mean age, 67 years) with peripancreatic masses were evaluated by EUS-FNA. All patients had initial computed tomography scanning with a mass seen in 49 patients, "fullness" to the pancreas in 28 patients, and no mass seen in 21 patients. Evaluation with EUS-FNA revealed 22 benign lesions, 18 T2 masses, 37 T3 masses, 1 T4 mass, and 20 masses representing nonpancreatic tumors. Results of EUS-FNA of adjacent lymph nodes were positive in 27 patients. Twenty-seven patients had surgical resection or palliation permitting operative and pathologic staging. On comparison of EUS FNA staging with surgical staging, 12 patients were the same stage, 14 patients were upstaged, and 1 patient was downstaged. The remaining patients who did not have surgery have been followed up for a mean of 15 months. Overall accuracy of EUS-FNA for differentiating benign from malignant masses was 96%. CONCLUSIONS: Endoscopic ultrasound-guided fine needle aspiration is a useful technique for the evaluation of pancreatic masses. It is highly accurate for differentiating between benign and malignant lesions and for predicting T stage, but is limited for predicting nodal status. PMID- 10367875 TI - Image-guided surgery: preliminary feasibility studies of frameless stereotactic liver surgery. AB - BACKGROUND: Liver surgery can be difficult because there are few external landmarks defining hepatic anatomy and because the liver has significant vascularity. Although preoperative tomographic imaging (computed tomography or magnetic resonance imaging) provides essential anatomical information for operative planning, at present it cannot be used actively for precise localization during surgery. Interactive image-guided surgery involves the simultaneous real-time display of intraoperative instrument location on preoperative images (computed or positron-emission tomography or magnetic resonance imaging). Interactive image-guided surgery has been described for tumor localization in the brain (frameless stereotactic surgery) and allows for interactive use of preoperative images during resections or biopsies. HYPOTHESIS: The application of interactive image-guided surgery (IIGS) is feasible for hepatic procedures from a biomedical engineering standpoint. METHODS: We developed an interactive image-guided surgery system for liver surgery and tested a porcine liver model for tracking liver motion during insufflation; liver motion during respiration in open procedures in patients undergoing hepatic resection; and tracking accuracy of general surgical instruments, including a laparoscope and an ultrasound probe. RESULTS: Liver motion due to insufflation can be quantified; average motion was 2.5+/-1.4 mm. Average total liver motion secondary to respiration in patients was 10.8 +/-2.5 mm. Instruments of varying lengths, including a laparoscope, can be tracked to accuracies ranging from 1.4 to 2.1 mm within a 27-m3 (3 X 3 X 3-m) space. CONCLUSION: Interactive image-guided surgery appears to be feasible for open and laparoscopic hepatic procedures and may enhance future operative localization. PMID- 10367876 TI - Sporadic primary hyperparathyroidism in young patients: a separate disease entity? AB - HYPOTHESIS: Sporadic primary hyperparathyroidism (1 HPT) in young persons is thought to be extremely rare. The exact incidence is unknown and little is known of the characteristics of the disease. METHODS: From 1976 to 1998, 33 patients aged 19 years or younger underwent operation for sporadic 1 HPT at a single institution. Data were recorded regarding the clinical, surgical, pathologic, and biochemical aspects, as well as long-term patient follow-up. RESULTS: There were 17 male subjects and 16 female subjects ranging in age from 9 to 19 years (median age, 17 years). Thirty-one (94%) were symptomatic: 14 (42%) had renal stones, 9 (27%) had bone disease, 1 (3%) had pancreatitis, and 7 (21%) had vague nonspecific symptoms alone. The high incidence of symptoms was matched by correspondingly high biochemical values (mean serum calcium level, 3.02 mmol/L [12.1 mg/dL]) and large adenomas (mean weight, 967 mg). Five patients (15%) underwent exploration for persistent/recurrent 1 HPT. Thirty-one patients (94%) were normocalcemic postoperatively. One patient was temporarily hypocalcemic. No patient had vocal cord paralysis or paresis. Two patients developed recurrent disease in the mean follow-up period of 10.3 years. None have shown evidence of an inherited disorder. CONCLUSIONS: It appears that 1 HPT in young patients presents as a more severe disease, in terms of symptoms, biochemistry, and extent of pathologic findings. Physicians should be aware that 1 HPT does occur in young persons in a nonfamilial setting and that it may be responsible for a wide spectrum of symptoms. As in the adult population, 1 HPT is safely and effectively treated with surgical intervention. PMID- 10367878 TI - The acute inflammatory response and its regulation. AB - The acute inflammatory response is composed of an elaborate cascade of both proinflammatory and anti-inflammatory mediators. The balance between these mediators often determines the outcome after injury. In clinical scenarios, such as trauma or sepsis, there is often unregulated production of proinflammatory mediators that can cause multiple organ failure. Further understanding of the endogenous mechanisms that control the inflammatory response is needed to facilitate development of therapeutic options. In this review, we discuss the current knowledge of the mechanisms leading to development of acute inflammatory injury as well as the factors that regulate this response. PMID- 10367877 TI - Candida infections in critically ill trauma patients: a retrospective case control study. AB - HYPOTHESIS: We sought to determine whether the usual risk factors for fungal infections are applied to trauma patients. DESIGN: Case-control study. SETTING: American College of Surgeons Committee on Trauma-certified Level I trauma center in a tertiary care community hospital. PATIENTS: Screening of medical records of a consecutive sample of 459 patients aged 16 years or older admitted to an intensive care unit for 4 days or more from 1993 through 1996 identified 20 patients infected with Candida species. Two case controls for each were selected from the remaining patients using sex, age within 5 years, mechanism of injury, and best fit of first 4 Abbreviated Injury Scale scores; the Injury Severity Score and intensive care unit length of stay were also used if needed. INTERVENTIONS: None. RESULTS: Univariate analyses by t and chi2 tests showed significance (P<.05) for number of units of blood transfused in the first 24 hours after injury, gastrointestinal perforation, hemodialysis, and total parenteral nutrition. Steroids, fungal colonization, use of central venous catheters, Acute Physiology and Chronic Health Evaluation II score, mechanical ventilation for 3 days or more, and the number and duration of antibiotics were not significantly different. Logistic regression analysis showed that only total parenteral nutrition was an independent risk factor in this trauma population. CONCLUSION: Many of the classic risk factors for fungal infection in other populations are actually concomitants of injury severity and its requisite level of care in trauma patients. Hyperalimentation in persistently critically ill trauma patients significantly increases the risk of Candida infection. PMID- 10367879 TI - Proctectomy and coloanal anastomosis for rectal cancer. AB - Fueled by a greater understanding of pelvic physiology along with an improved comprehension of rectal cancer spread, we are now able to offer most patients restoration of intestinal continuity following oncologic proctectomy. Coloanal or ultralow colorectal anastomosis can be performed in most patients with midrectal cancers, provided that anal sphincter function is not impaired preoperatively. Functional results may be improved by construction of a colonic pouch with pouch anal anastomosis. Temporary fecal diversion, usually with a diverting loop ileostomy, may be prudent, especially in patients undergoing neoadjuvant chemoradiation. PMID- 10367880 TI - Anesthesia during the Civil War. PMID- 10367882 TI - Dystroglycan versatility. PMID- 10367881 TI - Nuclear orphan receptors control cholesterol catabolism. PMID- 10367883 TI - Unlocking family secrets: K+ channel transmembrane domains. PMID- 10367884 TI - Unified nomenclature for the semaphorins/collapsins. Semaphorin Nomenclature Committee. PMID- 10367885 TI - BiP acts as a molecular ratchet during posttranslational transport of prepro alpha factor across the ER membrane. AB - We have addressed the mechanism by which proteins are posttranslationally transported across the membrane of the yeast endoplasmic reticulum (ER). We demonstrate that BiP (Kar2p), a member of the Hsp70 family resident in the ER lumen, acts as a molecular ratchet during translocation of the secretory protein prepro-alpha factor through the channel formed by the Sec complex. Multiple BiP molecules associate with each translocation substrate following interaction with the J domain of the Sec63p component of the Sec complex. Bound BiP minimizes passive backward movements of the substrate through the channel, and BiP's subsequent dissociation results in a free polypeptide in the ER lumen. Antibodies against the substrate can replace BiP, indicating that a Brownian ratchet is sufficient to achieve translocation. PMID- 10367886 TI - The protein import motor of mitochondria: unfolding and trapping of preproteins are distinct and separable functions of matrix Hsp70. AB - Mitochondrial heat shock protein 70 (mtHsp70) functions in unfolding, translocation, and folding of imported proteins. Controversial models of mtHsp70 action have been discussed: (1) physical trapping of preproteins is sufficient to explain the various mtHsp70 functions, and (2) unfolding of preproteins requires an active motor function of mtHsp70 ("pulling"). Intragenic suppressors of a mutant mtHsp70 separate two functions: a nonlethal folding defect caused by enhanced trapping of preproteins, and a conditionally lethal unfolding defect caused by an impaired interaction of mtHsp70 with the membrane anchor Tim44. Even enhanced trapping in wild-type mitochondria does not generate a pulling force. The motor function of mtHsp70 cannot be explained by passive trapping alone but includes an essential ATP-dependent interaction with Tim44 to generate a pulling force and unfold preproteins. PMID- 10367887 TI - Induction of GADD45 and JNK/SAPK-dependent apoptosis following inducible expression of BRCA1. AB - The breast cancer susceptibility gene BRCA1 encodes a protein implicated in the cellular response to DNA damage, with postulated roles in homologous recombination as well as transcriptional regulation. To identify downstream target genes, we established cell lines with tightly regulated inducible expression of BRCA1. High-density oligonucleotide arrays were used to analyze gene expression profiles at various times following BRCA1 induction. A major BRCA1 target is the DNA damage-responsive gene GADD45. Induction of BRCA1 triggers apoptosis through activation of c-Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK), a signaling pathway potentially linked to GADD45 gene family members. The p53-independent induction of GADD45 by BRCA1 and its activation of JNK/SAPK suggest a pathway for BRCA1-induced apoptosis. PMID- 10367888 TI - Reciprocal interactions of Pit1 and GATA2 mediate signaling gradient-induced determination of pituitary cell types. AB - The mechanisms by which transient gradients of signaling molecules lead to emergence of specific cell types remain a central question in mammalian organogenesis. Here, we demonstrate that the appearance of four ventral pituitary cell types is mediated via the reciprocal interactions of two transcription factors, Pit1 and GATA2, which are epistatic to the remainder of the cell type specific transcription programs and serve as the molecular memory of the transient signaling events. Unexpectedly, this program includes a DNA binding independent function of Pit1, suppressing the ventral GATA2-dependent gonadotrope program by inhibiting GATA2 binding to gonadotrope- but not thyrotrope-specific genes, indicating that both DNA binding-dependent and -independent actions of abundant determining factors contribute to generate distinct cell phenotypes. PMID- 10367889 TI - Cytonemes: cellular processes that project to the principal signaling center in Drosophila imaginal discs. AB - Wing imaginal disc cells in Drosophila develop by using information received from a signaling center associated with the anterior/posterior compartment border. We show here that disc cells have thin, actin-based extensions (cytonemes) that project to this signaling center. Cytonemes can be induced when cells from the lateral flanks of a wing disc are cultured next to cells from the A/P border or next to a source of fibroblast growth factor. Mouse limb bud cells also grow projections during a brief culture period, indicating that cytonemes are an attribute of both vertebrate and invertebrate cells. We suggest that cytonemes may be responsible for some forms of long-range cell-cell communication. PMID- 10367890 TI - MEC1-dependent redistribution of the Sir3 silencing protein from telomeres to DNA double-strand breaks. AB - The yeast Sir2/3/4p complex is found in abundance at telomeres, where it participates in the formation of silent heterochromatin and telomere maintenance. Here, we show that Sir3p is released from telomeres in response to DNA double strand breaks (DSBs), binds to DSBs, and mediates their repair, independent of cell mating type. Sir3p relocalization is S phase specific and, importantly, requires the DNA damage checkpoint genes MEC1 and RAD9. MEC1 is a homolog of ATM, mutations in which cause ataxia telangiectasia (A-T), a disease characterized by various neurologic and immunologic abnormalities, a predisposition for cancer, and a cellular defect in repair of DSBs. This novel mode by which preformed DNA repair machinery is mobilized by DNA damage sensors may have implications for human diseases resulting from defective DSB repair. PMID- 10367891 TI - Relocalization of telomeric Ku and SIR proteins in response to DNA strand breaks in yeast. AB - Telomeric TG-rich repeats and their associated proteins protect the termini of eukaryotic chromosomes from end-to-end fusions. Associated with the cap structure at yeast telomeres is a subtelomeric domain of heterochromatin, containing the silent information regulator (SIR) complex. The Ku70/80 heterodimer (yKu) is associated both with the chromosome end and with subtelomeric chromatin. Surprisingly, both yKu and the chromatin-associated Rap1 and SIR proteins are released from telomeres in a RAD9-dependent response to DNA damage. yKu is recruited rapidly to double-strand cuts, while low levels of SIR proteins are detected near cleavage sites at later time points. Consistently, yKu- or SIR deficient strains are hypersensitive to DNA-damaging agents. The release of yKu from telomeric chromatin may allow efficient scanning of the genome for DNA strand breaks. PMID- 10367892 TI - Structural view of the Ran-Importin beta interaction at 2.3 A resolution. AB - Transport receptors of the Importin beta family shuttle between the nucleus and cytoplasm and mediate transport of macromolecules through nuclear pore complexes. They interact specifically with the GTP-binding protein Ran, which in turn regulates their interaction with cargo. Here, we report the three-dimensional structure of a complex between Ran bound to the nonhydrolyzable GTP analog GppNHp and a 462-residue fragment from Importin beta. The structure of Importin beta shows 10 tandem repeats resembling HEAT and Armadillo motifs. They form an irregular crescent, the concave site of which forms the interface with Ran triphosphate. The importin-binding site of Ran does not overlap with that of the Ran-binding domain of RanBP2. PMID- 10367893 TI - Two structural transitions in membrane pore formation by pneumolysin, the pore forming toxin of Streptococcus pneumoniae. AB - The human pathogen Streptococcus pneumoniae produces soluble pneumolysin monomers that bind host cell membranes to form ring-shaped, oligomeric pores. We have determined three-dimensional structures of a helical oligomer of pneumolysin and of a membrane-bound ring form by cryo-electron microscopy. Fitting the four domains from the crystal structure of the closely related perfringolysin reveals major domain rotations during pore assembly. Oligomerization results in the expulsion of domain 3 from its original position in the monomer. However, domain 3 reassociates with the other domains in the membrane pore form. The base of domain 4 contacts the bilayer, possibly along with an extension of domain 3. These results reveal a two-stage mechanism for pore formation by the cholesterol binding toxins. PMID- 10367894 TI - Crystal structure of a phosphatidylinositol 3-phosphate-specific membrane targeting motif, the FYVE domain of Vps27p. AB - Phosphatidylinositol 3-phosphate regulates membrane trafficking and signaling pathways by interacting with the FYVE domains of target proteins. The 1.15 A structure of the Vps27p FYVE domain reveals two antiparallel beta sheets and an alpha helix stabilized by two Zn2+-binding clusters. The core secondary structures are similar to a rabphilin-3A Zn2+-binding domain and to the C1 and LIM domains. Phosphatidylinositol 3-phosphate binds to a pocket formed by the (R/K)(R/K)HHCR motif. A lattice contact shows how anionic ligands can interact with the phosphatidylinositol 3-phosphate-binding site. The tip of the FYVE domain has basic and hydrophobic surfaces positioned so that nonspecific interactions with the phospholipid bilayer can abet specific binding to phosphatidylinositol 3-phosphate. PMID- 10367895 TI - The chosen few? Positive selection and the generation of naive B lymphocytes. PMID- 10367896 TI - Developmental regulation of TCR delta locus accessibility and expression by the TCR delta enhancer. AB - We have used gene-targeted mutation to assess the role of the T cell receptor delta (TCR delta) enhancer (E delta) in alphabeta and gammadelta T cell development. Mice lacking E delta exhibited no defects in alphabeta T cell development but had a severe reduction in thymic and peripheral gammadelta T cells and decreased VDJ delta rearrangements. Simultaneous deletion of both E delta and the TCR alpha enhancer (E alpha) demonstrated that residual TCR delta rearrangements were not driven by E alpha, implicating additional elements in TCR delta locus accessibility. Surprisingly, while deletion of E delta severely impaired germline TCR delta expression in double-negative thymocytes, absence of E delta did not affect expression of mature delta transcripts in gammadelta T cells. We conclude that E delta has an important role in TCR delta locus regulation at early, but not late, stages of gammadelta T cell development. PMID- 10367897 TI - CBP/p300 integrates Raf/Rac-signaling pathways in the transcriptional induction of NF-ATc during T cell activation. AB - NF-ATc, an inducibly expressed transcription factor, controls gene expression in T lymphocytes and cardiomyocytes. We show here that the transcriptional co activators CBP/p300 bind to and control the activity of the inducible N-terminal transactivation domain of NF-ATc, TAD-A. Similar to the N terminal transactivation domain of c-Jun, TAD-A is inducibly phosphorylated, but this phosphorylation is dispensable for the interaction with CBP/p300. Constitutive active versions of c-Raf and Rac synergistically enhance the CBP/p300-mediated increase of TAD-A activity, indicating the important role CBP/p300 plays in the integration of T cell activation signals. Since a mutation of CBP abolishing HAT activity is almost as active as wild-type CBP in T cells, functions of CBP/p300 other than histone acetylation appear to control the NF-AT-dependent transcription in T cells. PMID- 10367898 TI - Distinct roles of the phosphatidylinositol 3-kinase and STAT5 pathways in IL-7 mediated development of human thymocyte precursors. AB - Here, we define the IL-7R-activated signal that promotes survival and proliferation of T cell progenitors and demonstrate that it is distinct from the signals that induce differentiation. We show that IL-7 activates PKB and STAT5 in human thymocytes. Into T cell precursors we introduced chimeric receptors with a cytoplasmic domain of the IL-7R that is no longer able to activate PI-3K/PKB and STAT5 and tested the transduced cells in a fetal thymic organ culture. We also examined the T cell precursor activity of progenitors expressing dominant negative forms of PI-3K or STAT5B. These experiments revealed that PI-3K/PKB activation is essential for the survival and proliferation of T cell precursors and suggest that STAT5 activated by IL-7 mediates T cell differentiation. PMID- 10367899 TI - Immature thymocytes employ distinct signaling pathways for allelic exclusion versus differentiation and expansion. AB - T cell receptor (TCR) beta chain allelic exclusion occurs at the thymocyte CD4- 8 (double-negative, or DN) to CD4+ 8+ (double-positive, or DP) transition, concurrently with differentiation and cellular expansion, and is imposed by a negative feedback loop in which a product of the first rearranged TCRbeta allele arrests further recombination in the TCRbeta locus. All of the major events associated with the development of DP cells can be induced by the introduction of TCRbeta or activated Lck transgenes. Here, we present evidence that the signaling pathways that promote thymocyte differentiation and expansion of RAG-deficient DN cells but not those that suppress rearrangements of endogenous TCRbeta genes in normal DN cells are engaged by activated Ras. We propose that TCRbeta allelic exclusion is mediated by effector pathways downstream of Lck but independent of Ras. PMID- 10367900 TI - Deficient T cell fate specification in mice with an induced inactivation of Notch1. AB - Notch proteins are cell surface receptors that mediate developmental cell specification events. To explore the function of murine Notch1, an essential portion of the gene was flanked with loxP sites and inactivation induced via interferon-regulated Cre recombinase. Mice with a neonatally induced loss of Notch1 function were transiently growth retarded and had a severe deficiency in thymocyte development. Competitive repopulation of lethally irradiated wild-type hosts with wild-type- and Notch1-deficient bone marrow revealed a cell autonomous blockage in T cell development at an early stage, before expression of T cell lineage markers. Notch1-deficient bone marrow did, however, contribute normally to all other hematopoietic lineages. These findings suggest that Notch1 plays an obligatory and selective role in T cell lineage induction. PMID- 10367901 TI - Designing and maintaining the mature TCR repertoire: the continuum of self peptide:self-MHC complex recognition. AB - Peripheral T cell maintenance requires a survival signal delivered upon T cell receptor (TCR)-major histocompatibility complex (MHC) molecule interaction. Since self-peptides play a critical role in the intrathymic positive selection of the mature TCR repertoire, we hypothesized an equally important role in T cell persistence. We used mice with a normal expression of MHC class II molecules but a restricted self-peptide complexity (H-2M alpha-/-) to show that an MHC class II restricted T cell specificity that displays a deficient positive selection in the H-2M alpha-/- thymus shows an impaired persistence after adoptive transfer in H 2M alpha-/- recipients. Finally, a wild-type CD4+ TCR repertoire is incompletely maintained in H-2M alpha-/- recipients. These observations suggest that, similar to intrathymic positive selection, the maintenance of the mature TCR repertoire relies on the recognition of self-peptide:self-MHC complexes. PMID- 10367902 TI - Generation of functional thymocytes in the human adult. AB - Reconstituting the immune response will be critical for the survival of HIV infected individuals once viral load is brought under control. While the adult thymus was previously thought to be relatively inactive, new data suggest it may play a role in T cell reconstitution. We examined thymopoiesis in adults up to 56 years of age and found active T cell receptor (TCR) rearrangement, generating a diverse TCR Vbeta repertoire. The resulting thymocytes are functional and are capable of responding to costimulatory signals. These data demonstrate that the adult thymus remains active late in life and contributes functional T cells to the peripheral lymphoid pool. PMID- 10367903 TI - Crystal structure of the MHC class I homolog MIC-A, a gammadelta T cell ligand. AB - The major histocompatibility complex (MHC) class I homolog MIC-A functions as a stress-inducible antigen that is recognized by a subset of gammadelta T cells independent of beta2-microglobulin and bound peptides. Its crystal structure reveals a dramatically altered MHC class I fold, both in detail and overall domain organization. The only remnant of a peptide-binding groove is a small cavity formed as the result of disordering a large section of one of the groove defining helices. Loss of beta2-microglobulin binding is due to a restructuring of the interaction interfaces. Structural mapping of sequence variation suggests potential receptor binding sites on the underside of the platform on the side opposite of the surface recognized by alphabeta T cell receptors on MHC class I peptide complexes. PMID- 10367904 TI - Granzyme A loading induces rapid cytolysis and a novel form of DNA damage independently of caspase activation. AB - Cytotoxic lymphocytes trigger apoptosis by releasing perforin and granzymes (Grn). GrnB activates the caspase apoptotic pathway, but little is known about GrnA-induced cell death. Perforin was used to load recombinant GrnA and GrnB and enzymatically inactive variants into target cells. GrnA induces single-strand DNA breaks that can be labeled with Klenow polymerase and visualized on alkaline gels. GrnA-induced DNA damage but not cytolysis requires GrnA proteolysis. GrnA induced membrane perturbation, nuclear condensation, and DNA damage are unimpaired by caspase blockade. GrnA fails to induce cleavage of caspase-3, lamin B, rho-GTPase, or PARP. GrnA-induced cytotoxicity and cleavage of PHAP II, a previously identified GrnA substrate, are unimpaired in Jurkat cells that overexpress bcl-2. Therefore, GrnA activates a novel apoptotic pathway. PMID- 10367905 TI - Granzyme A initiates an alternative pathway for granule-mediated apoptosis. AB - Granzyme (gzm) B-deficient cytotoxic lymphocytes (CTL) have a severe defect in the rapid induction of target cell apoptosis that is almost completely corrected by prolonged incubation of the CTL effectors and their targets. We show in this report that perforin-dependent, gzmB-independent cytotoxicity is caused by gzmA (or tightly linked genes). CTL deficient for gzmA and gzmB retain normal perforin function, but these CTL have a cytotoxic defect in vivo that is as severe as perforin-deficient CTL. Collectively, these results suggest that perforin provides target cell access and/or trafficking signals for the gzms, and that the gzms themselves deliver the lethal hits. The gzmA pathway appears to function independently from gzmB and may therefore provide a critical "back-up" system when gzmB is inhibited in the target cell. PMID- 10367906 TI - Increased junctional diversity in fetal B cells results in a loss of protective anti-phosphorylcholine antibodies in adult mice. AB - Fetal Igs are less diverse than adult Igs, largely because of the lack of N addition in the absence of Tdt. To test whether the absence of Tdt is essential, we generated Tg mice that express Tdt and add N regions in fetal B cells. When challenged as adults with PC-containing Streptococcus pneumoniae, these mice fail to make the hallmark T15 anti-PC Ab encoded by canonical rearrangements of Ig H and L chain genes. The anti-PC Abs from these mice are altered by premature N addition and do not protect against death from virulent pneumococcal infection. These results show that maintenance of lower Ig diversity in early life is essential for the acquisition of a complete functional adult repertoire. PMID- 10367907 TI - Mutations affecting either generation or survival of cells influence the pool size of mature B cells. AB - The mature B cell compartment of MHC class II-deficient B6 I-Aalpha(-/-) and the btk-defective CBA/N mouse strain is 4- to 5-fold smaller than in wild-type B6 mice. The defect in B6 I-Aalpha(-/-) mice is intrinsic to B cells and due to a 4- to 5-fold reduced lifespan, which however can be normalized by an I-Ealpha(d) transgene, but only when expressed early during B cell development. The reduced number of mature B cells in the btk-defective CBA/N mouse is due to a 4- to 5 fold lower number of immature splenic B cells entering the mature compartment. The combined defects of reduced lifespan and impaired generation in double mutant mice result in a severe deficiency in the mature B cell pool. PMID- 10367908 TI - Novel treatment strategies for superficial mycoses: introduction. PMID- 10367909 TI - Diagnosis of onychomycosis made simple. AB - Because onychomycosis requires long-term systemic therapy, it is essential to diagnose the infection accurately. Although in theory this diagnosis should be simple, results achieved in practice are uneven. Clinicians should be aware of the need to collect an adequate specimen and of the possible need for repeat collections. Direct microscopic techniques for examination of nail scrapings include 10% potassium hydroxide (KOH) with Parker ink, 10% KOH with dimethyl sulfoxide, and 10% KOH with Calcofluor white. When interpreting fungal culture results, it should be noted that negative results are frequent and that contaminant yeasts and molds are common, but that nondermatophytes such as Candida, Scopulariopsis, and Scytalidium can infrequently cause onychomycosis. PMID- 10367910 TI - Optimal growth conditions for the determination of the antifungal susceptibility of three species of dermatophytes with the use of a microdilution method. AB - As a prerequisite to standardization of dermatophyte susceptibility testing, conditions that support optimal growth of different dermatophyte species must be established. Eighteen isolates of Trichophyton spp. (T rubrum, T mentagrophytes, T tonsurans) were grown in 4 different media: RPMI 1640 with L-glutamine, without sodium bicarbonate and buffered at pH = 7.0; antibiotic medium #3 (Penassay); yeast nitrogen base with 0.5% dextrose buffered at pH = 7.0; and Sabouraud dextrose broth. Incubation for 6 days at 35 degrees C produced the following results: RPMI and Sabouraud dextrose supported equally sufficient growth for all strains tested; Penassay supported growth of only 33% of the isolates tested, and buffered yeast nitrogen base did not support growth of any isolates. RPMI was selected as the optimal medium, and organisms were tested at both 30 degrees C and 35 degrees C with a standardized inoculum density of 10(3) conidia/mL. No temperature differences were noted in the amount of growth of the dermatophytes tested. With RPMI at an incubation temperature of 35 degrees C, 3 inoculum sizes (10(3), 10(4), and 10(5) conidia/mL) were tested against 4 antifungal agents: griseofulvin, itraconazole, terbinafine, and fluconazole. Inoculum size did not affect minimum inhibitory concentration (MIC) results for itraconazole or terbinafine, but a larger inoculum produced a slightly higher MIC for griseofulvin and a noticeably higher MIC for fluconazole. Our data support the use of RPMI 1640, 35 degrees C, and 4 days as an incubation temperature and time, respectively, and an inoculum of 10(3) conidia/mL as optimal conditions for the determination of the antifungal susceptibility of dermatophytes. PMID- 10367911 TI - Pharmacokinetics of fluconazole in skin and nails. AB - Two studies on the pharmacokinetics of fluconazole in skin and nails are reported here. In 1 study, 12 healthy volunteers received fluconazole 50 mg once daily for 12 days and 11 healthy volunteers received fluconazole 150 mg once weekly for 2 weeks. Fluconazole assays were performed on samples of serum, stratum corneum, dermis-epidermis, and eccrine sweat. In a second study, 36 patients with toenail onychomycosis received either fluconazole 150 mg once weekly or griseofulvin 1000 mg once daily for 12 months. Fluconazole assays were performed on nail clippings and serum samples from the patients receiving fluconazole. Tissue concentrations of fluconazole regularly exceeded plasma concentrations in these studies. In the skin study, the highest concentrations were achieved in stratum corneum, with accumulation occurring up to the end of dosing. Subjects who received 50 mg once daily had higher levels of fluconazole in stratum corneum, sweat, and epidermis dermis than those subjects who received 150 mg once weekly. In the toenail study, fluconazole concentrations increased for the first 6 months, reaching levels much higher than serum concentrations (P < .001), with no significant difference between healthy and diseased nails. PMID- 10367912 TI - Onychomycosis: therapeutic update. AB - Onychomycosis is a common disease of the nail unit caused by dermatophytes, yeasts, and molds. In more than 80% of cases, onychomycosis is caused by the dermatophytes Trichophyton rubrum and Trichophyton mentagrophytes. The prevalence of onychomycosis in the world's population is 2% to 18% or higher and accounts for approximately 50% of all nail disorders. Until recently, available therapies were inadequate because of low cure rates, high relapse rates, and often dangerous side effects. An increased understanding of nail pharmacokinetics has led to the development of safer, more effective systemic therapies for onychomycosis, such as itraconazole, fluconazole, and terbinafine. These new oral antifungal agents allow shorter periods of treatment, provide rapid efficacy, and may improve patient compliance and attitudes regarding therapy. Treatment selection will depend on several factors, including appropriate spectrum of activity, adverse effects, and potential drug interactions plus patient preferences for specific dosing regimens. PMID- 10367913 TI - Treatment of tinea capitis: beyond griseofulvin. AB - Tinea capitis is a common pediatric scalp infection caused by dermatophytes. Topical therapy alone is ineffective, so oral griseofulvin has traditionally been the standard treatment. The new antimycotic agents itraconazole, terbinafine, and fluconazole represent effective treatment alternatives that have fewer problems with tolerability and adverse effects. More comparative studies are needed to determine the optimal treatment with these agents and adjuvant therapies such as antifungal shampoos, topical antimycotic agents, and corticosteroids. PMID- 10367914 TI - Oral therapy of common superficial fungal infections of the skin. AB - Although superficial fungal infections of the skin often respond to topical agents, systemic therapy is sometimes necessary. This article gives a review of the effectiveness of the oral antifungal agents fluconazole, itraconazole, and terbinafine in the treatment of pityriasis versicolor, tinea corporis/cruris, and tinea pedis. Four hundred milligrams fluconazole as a single dose and 200 mg itraconazole daily for 5 to 7 days were effective in the treatment of pityriasis versicolor; terbinafine taken orally appears to be ineffective in pityriasis versicolor. Tinea corporis and tinea cruris were effectively treated by 50 to 100 mg fluconazole daily or 150 mg once weekly for 2 to 3 weeks, by 100 mg itraconazole daily for 2 weeks or 200 mg daily for 7 days, and by 250 mg terbinafine daily for 1 to 2 weeks. Tinea pedis has been effectively treated with pulse doses of 150 mg fluconazole once weekly, with 100 mg itraconazole daily for 2 weeks or 400 mg daily for 1 week, and with 250 mg terbinafine daily for 2 weeks. PMID- 10367915 TI - The management of superficial candidiasis. AB - Superficial Candida infection includes several common conditions, most often related to some underlying local or systemic predisposition. Appropriate identification of the pathogen is important in the management of candidiasis as the result of differences in susceptibility among species and strains of Candida to different antifungal drugs. Treatment options are reviewed for oropharyngeal candidiasis, vaginal candidiasis, cutaneous candidiasis, paronychia and onychomycosis, and chronic mucocutaneous candidiasis. Because of the importance of predisposing conditions for candidiasis, adjunctive measures to abate these may be useful, although they are seldom effective in immunocompromised patients. PMID- 10367916 TI - Mal de debarquement. AB - MAIN OUTCOME MEASURE: Clinical features of mal de debarquement syndrome. RESULTS: Nearly all respondents were middle-aged women (26 of 27; mean age, 49.3 years). The duration of symptoms ranged from 6 months to 10 years (mean, 3.5 years; SD, 2.5 years). The symptoms were constant in 23 (85%) patients. Neither meclizine hydrochloride nor transdermal scopolamine was helpful. Benzodiazepines were of the most benefit. Balance rehabilitation physical therapy was undertaken by 15 patients, who on average reported a small benefit. CONCLUSIONS: More than double the number of previously reported cases of mal de debarquement syndrome were identified by this study. The syndrome usually occurs in middle-aged women following an ocean cruise. Symptoms are often refractory to vestibular suppressants as well as physical therapy. PMID- 10367917 TI - Design and test of a new tracheostoma valve based on inhalation. AB - BACKGROUND: Tracheostoma valves are used to make hand-free speaking possible for persons who have undergone a laryngectomy. OBJECTIVE: To design and test a new tracheostoma valve to improve existing tracheostoma valves. METHODS: The tracheostoma valve closes by means of strong inhalation so that all the air that is exhaled is available for phonation. The device automatically stays in the"speaking position" until the patient deliberately changes the device to the "breathing position" by a fast expiration. If all the air that has been exhaled has been consumed during phonation, the patient can inhale again, without changing the device, because a small valve automatically opens, thus allowing phonation without time limits. An experimental setup with a computer-based acquisition program was used to measure the pressure at which the valve opened and the flow at which the valve closed. The pressure and flow needed to open and close the magnetic adjustable valve were measured for different positions and contained in the computer through a data acquisition program. Also, the airflow resistance coefficients for inhaling and exhaling were measured. RESULTS: The airflow necessary to close the tracheostoma valve ranges from 1.6 to 3.8 L/s. The opening pressure of the valve ranges from 1 to 7 kPa. The airflow resistance coefficient is 290 Pa x s2 x L(-2) for inhalation and 430 Pa x s(2) x L(-2) for exhalation. CONCLUSIONS: The device appears to function well in physiological ranges and is optimally adjustable. The airflow resistance coefficient lies in the range of the entire airway resistance (120-470 Pa x s(2) x L(-2)) in quiet breathing. PMID- 10367918 TI - Long-term enhancement of botulinum toxin injections by upper-eyelid surgery in 14 patients with facial dyskinesias. AB - OBJECTIVES: To determine the effects of upper-eyelid surgery (limited myectomy, blepharoplasty, and levator aponeurotic advancement) on patients who demonstrated a suboptimal response or residual heaviness of the upper eyelids after botulinum toxin eyelid injections for facial dyskinesia. DESIGN: Retrospective study. SUBJECTS: Charts of 358 patients with a diagnosis of benign essential blepharospasm, Meige syndrome (with eyelid involvement), and hemifacial spasm were reviewed. METHODS: Data were retrospectively analyzed and included subjective and objective responses about botulinum toxin injections (number and duration of effect of injections before and after eyelid surgery). RESULTS: Of 358 patients with facial dyskinesias, 14 (3.91%) underwent upper-eyelid limited myectomy with or without upper-lid blepharoplasty (n = 5), upper-lid blepharoplasty alone (n = 6), or levator advancement with or without blepharoplasty (n = 3). Mean subjective improvement was 68.75% after limited myectomy combined with blepharoplasty and 58.33% after levator and/or blepharoplasty surgery. Average duration of effect of injections increased from 122.1 days in the patients prior to undergoing eyelid surgery to 210.5 days after surgery. CONCLUSIONS: Upper-eyelid surgery, including limited myectomy, enhanced the effect of the botulinum toxin in this small group of patients. Patients with a suboptimal response to injections in terms or moderate to marked dermatochalasis with subjective heaviness of the eyelids, upper-eyelid blepharoplasty, and/or limited myectomy should be considered. PMID- 10367920 TI - Total, subtotal, and partial surgical removal of cervicofacial lymphangiomas. AB - OBJECTIVES: To compare different surgical interventions for the treatment of extensive cervicofacial lymphangiomas and to define the minimal extent of surgery necessary to control disease. DESIGN: Retrospective study. Mean +/- SD follow-up was 31+/-4 months after surgery. Surgical procedures were grouped as follows: (1) total removal, (2) subtotal removal (all cystic structures removed, small plaques of cyst walls left attached to vital structures), (3) partial removal (major cysts removed, some partially resected cystic structures left in place), and (4) incision and aspiration with subsequent compression bandage. Control of disease was defined as no recurrent or residual tumor or as recurrent or residual tumor less than 10% of initial tumor size without evidence of growth on several postoperative examinations and without clinical symptoms or aesthetic disfigurement. PATIENTS: Twenty-one patients with cervicofacial lymphangiomas (>3 cm in maximum diameter) without thoracic involvement were evaluated. Fifteen patients were 6 years or younger and 6 were older than 6 years. No surgery was yet performed in 3 patients, for a total of 24 surgical interventions in 18 patients. SETTING: Hospitalized care in 2 referral centers. RESULTS: After total removal, disease was controlled in 5 of 5 cases; after subtotal removal, in 8 of 9 cases; after partial removal, in 1 of 7 cases; and after incision and aspiration with subsequent compression bandage, in 0 of 3 cases. Two complications were encountered-1 fully reversible paresis of the marginal branch of the facial nerve and 1 secondary healing. CONCLUSIONS: Surgical removal of cervicofacial lymphangiomas is a safe treatment modality. Disease control can be achieved if all cystic structures are removed. Small plaques of cyst walls attached to vital structures may be left in place. If small cystic extensions of lymphangiomas are only opened and left in place or if lymphangiomas are only drained following compression bandage, symptomatic residual tumor or recurrence is frequent. PMID- 10367919 TI - Malignant melanomas of the parotid: comparison of survival for patients with metastases from known vs unknown primary tumor sites. AB - BACKGROUND: Malignant melanoma (MM) rarely affects the parotid, and usually this diagnosis will herald a search for a primary skin neoplasm. Occasionally, no primary tumor is ever found, raising questions regarding prognosis and the issue of primary melanoma of the parotid. OBJECTIVE: To evaluate retrospectively the clinical and histological features of MM involving the parotid in 19 patients. DATA SOURCES: Pathology and hospital files at 3 tertiary care university hospitals. STUDY SELECTION: Patients with MM within the parotid with adequate histopathologic and immunohistochemical documentation, as well as clinical information regarding patient outcome. DATA EXTRACTION: In 6 patients, no extraparotid MM was ever identified. After parotidectomy, 5 patients (including 1 patient who died of other causes) were melanoma free at a mean of 4.2 years (range, 14 months to 7.5 years). Only 1 patient died of disease after 17 months. An extraparotid primary tumor was present in 13 patients, 10 with dermal and 3 with mucosal sites. At follow-up, only 1 of these patients was disease free after 2 years. Nine patients died of melanoma after a mean of 2.6 years (range, 10 months to 5 years); the other 3 had evidence of metastatic disease at a mean of 4.3 years (range, 3-6 years). Nondermal sites of primary tumors were the nasal cavity, sclera, and conjunctiva. DATA SYNTHESIS: Patients with metastatic MM from unknown primary tumors have a longer disease-free survival than those with metastases from known primary disease. CONCLUSIONS: Although rare, MM should be considered in the differential diagnosis of parotid tumors. Unusual mucosal or ocular sites should be considered in the search for possible primary tumor sites to avoid treatment delay. These data support the idea that patients with metastatic MM from unknown primary tumors may follow a more improved course than that of patients with metastases from known primary disease. PMID- 10367921 TI - Pitfalls in laryngotracheal reconstruction. AB - OBJECTIVE: To determine the causes of laryngotracheal reconstruction (LTR) failures. DESIGN: Retrospective chart review. SETTING: Tertiary care children's hospital. PATIENTS: Seventeen pediatric patients who underwent revision LTR from October 1, 1986, to December 31, 1998. INTERVENTION: Laryngotracheal reconstruction. MAIN OUTCOME MEASURE: Decannulation. RESULTS: Seventeen patients required a total of 42 LTRs for decannulation. There were 17 primary LTRs and 25 revision LTRs. The primary LTRs were done either at our or other institutions. Two patients died after initial LTR failed, one because of tracheotomy tube plugging and the other because of a severe respiratory syncytial virus pneumonia. All 15 remaining patients have been decannulated. There were 27 failed LTRs with 17 being primary and 10 revision LTR failures. In 3 of the 27 failed procedures, no obvious causes for failure could be found. In the remaining 24 procedures, 1 or more factors that contributed to LTR failure could be found. Poor preoperative evaluation with subsequent failure to address the airway lesion was seen in 6 procedures. Intraoperative reasons for LTR failure included inappropriate choice of graft in 2 procedures; inappropriate stent in 7; inappropriate stent length in 1; and inappropriate duration of stent in 8. In 6 procedures, the airway abnormalities identified at endoscopy were not adequately addressed at LTR. Postoperative factors for failure were poor follow-up in 2, anterior suprastomal collapse in 2, and slipped or broken stent in 2. Other factors that contributed to LTR failures included intractable gastroesophageal reflux disease in 1 procedure and keloid formation in 5. CONCLUSIONS: Although some LTRs may fail secondary to factors that are not under the surgeon's control, many LTR failures can be avoided by accurate preoperative and intraoperative assessment of the stenosis, correct choice of surgical procedure, and close postoperative monitoring. PMID- 10367922 TI - Genetic associations in age-related hearing thresholds. AB - OBJECTIVE: To determine the inheritance of age-related hearing loss. DESIGN: Cohort study comparing aggregation of hearing levels in genetically unrelated people (spouse pairs) and in genetically related people (sibling pairs, parent child pairs). SETTING: Framingham Heart Study biennial Examination 15 (1973-1975) and Framingham Offspring Study Examination 6 (1995-1998). SUBJECTS: Members of the Framingham cohorts with hearing tests and with a relative in the Framingham hearing study. MAIN OUTCOME MEASURES: Audiometric pure-tone thresholds at 250 to 8000 Hz were obtained and pure-tone average (PTA) hearing thresholds were calculated for the middle (0.5-2 kHz), high (4-8 kHz), and low (0.25-1 kHz) frequencies for each ear. The shape of the audiogram was categorized as either normal, abrupt high-frequency loss (sensory phenotype) or flat loss (strial phenotype). Correlations were made using the Familial Correlations program of the Statistical Analysis for Genetic Epidemiology software system. The level of significance was P = .01. RESULTS: Hearing threshold levels did not aggregate in spouses. Significant aggregation was noted in siblings and parent-child pairings for PTA at low, middle, and high frequencies. Sisters but not brothers had significant aggregation of each PTA measure. Mother-daughter and mother-son pairs but not father-son pairs had significant aggregation of hearing levels. For the sensory phenotype, there was significant aggregation in all related pairs except for father-child pairs. For the strial phenotype, there was significant aggregation of hearing levels in the related female pairs but not in the related male pairs. CONCLUSIONS: A clear familial aggregation occurs for age-related hearing levels, sensory presbycusis phenotypes, and strial presbycusis phenotypes. The aggregations are stronger in women than in men. The heritability estimate was greater for the strial phenotypes than for the sensory phenotypes. The data support a genetic effect on the inheritance of presbycusis in women and a mixed, genetically acquired cause in men. PMID- 10367923 TI - Short tone burst-evoked myogenic potentials on the sternocleidomastoid muscle: are these potentials also of vestibular origin? AB - OBJECTIVES: To show that short tone bursts (STBs) evoke myogenic potentials from the sternocleidomastoid muscle (SCM) that are of vestibular origin. DESIGN: Evoked potential activity was recorded from the SCMs of normal volunteers and from patients with vestibulocochlear disorders. SETTING: This outpatient study was conducted at the Department of Otolaryngology, University of Tokyo, Tokyo, Japan. SUBJECTS: Nine normal volunteers and 30 patients (34 affected ears) with vestibulocochlear disorders were examined. INTERVENTION: Diagnostic. OUTCOME MEASURES: Sound-evoked myogenic potentials in response to STBs were recorded with surface electrodes over each SCM. Responses evoked by STBs in patients were compared with responses evoked by clicks. RESULTS: In all normal subjects, STBs (0.5, 1, and 2 kHz) evoked biphasic responses on the SCM ipsilateral to the stimulated ear; the same was true for clicks. Short tone bursts of 0.5 kHz evoked the largest responses, while STBs of 2 kHz evoked the smallest. In patients with vestibulocochlear disorders, responses to STBs of 0.5 kHz were similar to responses evoked by clicks. Thirty (88%) of the 34 affected ears demonstrated the same results with 0.5-kHz STBs and with clicks. Responses were present in patients with total or near-total hearing loss and intact vestibular function. Conversely, patients with preserved hearing but with absent or severely decreased vestibular function had absent or significantly decreased myogenic potentials evoked by STBs. CONCLUSIONS: Short tone bursts as well as clicks can evoke myogenic potentials from the SCM. Myogenic potentials evoked by STBs are also probably of vestibular origin. PMID- 10367924 TI - Considerations for free-flap reconstruction of the hard palate. AB - OBJECTIVE: To evaluate the use of microvascular free-tissue transfers in the reconstruction of hard palate defects. DESIGN: Retrospective review of a case series. SETTING: Two tertiary referral centers. PATIENTS: Thirty patients had hard palatal defects that resulted from ablative oncologic surgery: 10 total or subtotal palatal defects, 14 hemipalatal defects, and 6 anterior arch defects. INTERVENTION: Nine fibular, 11 rectus abdominus, 3 scapular, 6 radial forearm, and 1 latissimus dorsi free flaps were used to reconstruct these defects. MAIN OUTCOME MEASURES: Separation of the oral cavity from the nasal and sinus cavities, complications, oral diet, speech intelligibility, and overall quality of life. RESULTS: No flap failures occurred, and all palatal defects were ultimately sealed. Nineteen patients eat a regular diet, while the remainder maintain a soft diet. Twelve patients use a conventional dental prosthesis; 8 of the dental prostheses are supported by implants. Of 23 patients examined for speech, 18 have no disorders, 3 exhibit hyponasal speech, and 2 have hypernasal speech. Overall University of Washington, Seattle, quality of life scores were fair in 2 patients, good in 6, and excellent in 12. CONCLUSIONS: Free-flap reconstruction of the palate provides reliable permanent separation of the oral and sinonasal cavities in one stage. In addition, the potential for dental rehabilitation with the restoration of masticatory function and normal phonation exists. Flap choice is tailored to specific palatal defects as well as patient needs. PMID- 10367925 TI - Efficacy of targeted chemoradiation and planned selective neck dissection to control bulky nodal disease in advanced head and neck cancer. AB - PURPOSE: To determine the efficacy of targeted chemoradiation with the radiation plus platinum (RADPLAT) protocol and planned selective neck dissection in patients with N2 to N3 nodal disease associated with upper aerodigestive tract carcinoma. METHODS: Analysis of 52 patients with N2a, N2b, or N3 disease involving 60 heminecks treated with intraarterial cisplatin, 150 mg/m2, and intravenous sodium thiosulfate, 9 g/m2, on days 1, 8, 15, and 22; radiation therapy, 180 to 200 cGy per fraction for 35 fractions (total dose, 68-74 Gy); and planned neck dissection (33 of 35 procedures were selective). RESULTS: Of the 56 evaluable heminecks, a clinical complete response was achieved in 33 (59%). Within this group, 16 neck dissections were performed, none of which yielded disease on pathological examination. A clinical partial response was obtained in 21 heminecks, of which 18 subsequently had a neck dissection, yielding disease on pathological examination in 14. In all cases, it was possible to completely excise all adenopathy with clear margins on pathological examination. The rate of regional disease control among the 56 evaluable heminecks was 91% (51/56) (median follow-up, 36 months). Four failures were associated with uncontrolled disease at other sites, and 1 was an isolated neck recurrence. CONCLUSION: Selective neck dissection appears to be an effective adjunct to targeted chemoradiation in controlling N2 to N3 neck disease. PMID- 10367926 TI - Comparison of growth factor expression in fetal and adult fibroblasts: a preliminary report. AB - BACKGROUND: Fetal wounds can heal without any histological evidence of scarring. Fetal wounds lack the inflammatory infiltrate characteristic of adult wounds, and the fetal environment is not necessary for scarless healing to occur. Recent evidence suggests that fibroblasts are the main effector of scarless healing in fetal tissue. What has not been shown is what profile of growth factors the fibroblast uses to influence wound repair. OBJECTIVE: To determine the expression of growth factors (transforming growth factors beta1, beta2, and beta3; acidic and basic fibroblast growth factors; keratinocyte growth factor; and platelet derived growth factor AA, BB, and AB) of fetal and adult fibroblasts in vitro. DESIGN: Adult and fetal fibroblasts were grown in culture, and messenger RNA was extracted by standard techniques. Northern hybridization was used to identify messenger RNA transcripts for the aforementioned growth factors. Densitometry was used to compare growth factor messenger RNA expression with that of a ubiquitously expressed control, glyceraldehyde phosphate dehydrogenase. RESULTS: The data suggest that fetal and adult fibroblasts express acidic and basic fibroblast growth factor and transforming growth factor beta1. Adult fibroblasts show twice the relative expression of these growth factors compared with fetal fibroblasts. CONCLUSIONS: The adult fibroblasts demonstrate a relative excess production of cytokines compared with fetal fibroblasts. This is thought to contribute to suboptimal wound healing in adult wounds compared with the scarless healing of fetal wounds. PMID- 10367927 TI - Nitric oxide accumulation in the nonventilated nasal cavity. AB - BACKGROUND: Nasal nitric oxide is present in high concentrations in the upper airway relative to the lower respiratory tract. OBJECTIVE: To explore the rate of nitric oxide accumulation in the nonventilated nasal cavity. METHODS: In 9 healthy subjects previously trained to close the soft palate, steady-state plateau nitric oxide levels were recorded while air was aspirated through the nasal airway in series at a constant flow rate. Nitric oxide was then allowed to accumulate in the nasal cavity by occluding both nares and keeping the velum closed. After varying occlusion times, peak nitric oxide levels and a second plateau were ascertained. RESULTS: While the subjects aspirated air at a constant flow, there was a slow rise to a first nitric oxide plateau. On opening to the analyzer after the accumulation period, the peak nitric oxide level was several times higher than the initial plateau (range, 2810-19008 ppb) and then slowly returned to previous plateau levels. There was no significant difference between initial and second plateau nitric oxide levels for any period. The accumulated nitric oxide peak increased in direct proportion to the accumulation time (P<.001). CONCLUSIONS: Nitric oxide concentrations accumulate in the nonventilated nasal cavity in proportion to the time of nonventilation. Peak nasal nitric oxide values after accumulation are similar to published sinus nitric oxide measurements obtained by direct puncture. These results suggest an important alternative source of nitric oxide in humans. PMID- 10367928 TI - New modification of hot-water irrigation in the treatment of posterior epistaxis. AB - BACKGROUND: Tamponade treatment for epistaxis is painful and traumatic to the nasal mucosa, and may necessitate hospitalization for several days. Hot-water irrigation (HWI) was introduced as a treatment of epistaxis more than 100 years ago. In a previous study the treatment proved to be effective, less painful, and less traumatic, and required a shorter hospital stay than tamponade treatment. However, HWI has the risk of aspiration during treatment. To minimize this risk, a special catheter has been designed. OBJECTIVES: To evaluate the modified HWI and to compare the results with tamponade treatment, with respect to patient compliance, effectiveness, recurrence of bleeding, pain, complications, and length of hospital stay. PATIENTS: A total of 122 patients, hospitalized for posterior epistaxis, were randomized to receive either HWI or tamponade treatment. RESULTS: In the HWI group, 31 (55%) of the patients could be discharged from the hospital after the initial treatment only, compared with 29 (44%) of the patients treated with tamponade. Using a 10-cm visual analog scale, the mean pain score during treatment was 4.7 in the HWI group compared with 7.5 in the tamponade group. The mean hospital stay was 2.9 days for the HWI group vs. 4.0 days for the tamponade group. After discharge from the hospital, necrosis or synechia was found on rhinoscopy in 12 patients (40%) in the tamponade group compared with none in the HWI group. CONCLUSIONS: Compared with tamponade treatment, HWI is as effective, requires a significantly shorter hospital stay, is less traumatic to the nose, and is significantly less painful. PMID- 10367929 TI - Left vocal cord paralysis as a primary manifestation of invasive pulmonary aspergillosis in a nonimmunocompromised host. AB - We report the first case (to our knowledge) of vocal cord paralysis as a primary manifestation of invasive pulmonary aspergillosis, which occurred in a 69-year old woman without immunodeficiency. Her chest radiograph showed left upper lobe infiltration with pleural thickening, and a computed tomogram of her chest showed a thick pleural reaction and fibrosis around the arch of the aorta. A transbronchial biopsy specimen revealed Aspergillus infection. The patient was treated with oral itraconazole. However, since vocal cord paralysis persisted, the patient underwent type I thyroplasty to improve vocal function. A review of the literature showed that the incidence of invasive pulmonary aspergillosis has increased, even in nonimmunocompromised subjects, and that the disease has a potential for recurrent laryngeal nerve palsy. Therefore, invasive pulmonary aspergillosis should be considered in patients with vocal cord paralysis. PMID- 10367930 TI - Pathologic quiz case 1. Acinic cell carcinoma of the deep lobe of the parotid gland involving the right parapharyngeal space. PMID- 10367931 TI - Pathologic quiz case 2. Cervical thymus. PMID- 10367932 TI - A 3-year-old child with a severely deviated septum and airway obstruction. PMID- 10367934 TI - A conservative role for septoplasty in young children. PMID- 10367933 TI - Septoplasty in children--yes, but do the right thing. PMID- 10367935 TI - Does "death receptor" signaling play a role in tumorigenesis and cancer therapy? AB - Physiological cell death, known as apoptosis, is an evolutionarily conserved process that is required for normal development and function of multicellular organisms. Abnormalities in cell death control are implicated as a cause or contributing factor in a range of diseases, including cancer, autoimmunity, and degenerative disorders. Importantly, the propensity of a cell to undergo apoptosis is one of the determinants of the sensitivity of tumor cells to antineoplastic therapy. Apoptosis can be triggered by stress-induced signals that arise from within the doomed cell or by signals that are elicited by binding of extracellular "death ligands" to their "death receptors." Cysteine proteases have been recognized as essential effectors of all pathways to apoptosis. Experiments with transgenic mice and gene knockout mice have shown that different caspases and their adaptor molecules are needed for "death receptor" signaling and apoptotic pathways elicited by cytokine withdrawal, DNA damage, or corticosteroids. These differences allow the pathways to be regulated by distinct inhibitors. It has been published that chemotherapeutic drugs and gamma-radiation induce apoptosis by "death ligand"-mediated activation of "death receptors," but this model has been challenged. Our review discusses this controversy in the light of current knowledge of the molecular control of apoptosis. PMID- 10367937 TI - Antitumor activity of alpha-galactosylceramide, KRN7000, in mice with EL-4 hepatic metastasis and its cytokine production. AB - Liver metastasis of primary tumor is a clinically major problem. KRN7000, an alpha-galactosylceramide, significantly augments natural killer (NK) activity of spleen cells and shows strong antitumor activity in mice with lung metastasis of melanoma B16 cells. To test whether KRN7000 has an antitumor activity in mice with hepatic metastasis of tumors, we examined the effect of KRN7000 on NK activity of hepatic mononuclear cells (MNC) and the antitumor activity in mice with liver metastasis of EL-4 cells. The in vivo administration of KRN7000 significantly augmented NK activity of hepatic MNC and inhibited tumor growth of EL-4 cells in the liver more markedly than chemotherapeutic agents, leading to a relatively high rate of cured mice. In addition, it appeared that the KRN7000 treatment is effective in mice with established EL-4 tumors. Moreover, we found that KRN7000 can produce significant amounts of interleukin 2 (IL-2), IL-4, IL 12, and interferon-gamma in a dose-dependent manner. These results suggest that KRN7000 will be useful for the treatment of cancer liver metastasis. PMID- 10367936 TI - Persistent distension and enhanced diffusive extravasation of tumor vessels improved uniform tumor targeting of radioimmunoconjugate in mice administered with angiotensin II and kininase inhibitor. AB - Induced hypertension with angiotensin II (AT-II) and the inhibition of kininase with enalapril maleate may increase the tumor targeting of radiolabeled monoclonal antibodies (MAbs). We previously found that short-period infusion of 2.0 microg/kg/min of AT-II enhanced tumor targeting of MAb without an impact on normal tissue distribution. In this study, we aimed to optimize the manipulation with these agents, and examine the possible mechanism of their effects on MAb distribution. Effect of the manipulation on tissue circulation was assessed in mice bearing colon cancer xenografts by 201Tl and 99mTc-human serum albumin (HSA) as markers of tissue blood flow and tissue blood volume and/or vascular permeability. A dose finding study of enalapril ranging from 3 to 300 microg showed that 30 microg of enalapril in combination with AT-II infusion produced the best improvement in tumor uptake of 99Tc-HSA without altering 201Tl distribution, suggesting that the increase of vascular permeability was caused by enalapril. AT-II infusion for longer than 1 h affected renal blood flow and caused subcutaneous edema. Tumor uptake of (111)In-A7, a murine IgG1, was 1.62 fold improved 72 h postinjection (P < 0.001) and intratumoral distribution became uniform with 2.0 microg/kg/min of AT-II for 1 h and 30 microg of enalapril. Vessels in manipulated tumors were distended even 48 h after the cessation of AT II infusion. In conclusion, it was suggested that persistent distension of tumor vessels and the increase of diffusive extravasation of MAb caused by short-period induced hypertension and inhibition of bradykinin degradation produced favorable effect for the MAb distribution in tumors. PMID- 10367938 TI - Establishment of a lymph node metastatic model of mouse hepatocellular carcinoma Hca-F cells in C3H/Hej mice. AB - To establish a transplantable lymph node metastatic animal model, mouse hepatocellular carcinoma Hca-F cells were implanted into C3H/Hej mice by subcutaneous inoculation. The characters of the metastasis were determined macroscopically and microscopically. During 17 generations, the percentages of metastasis of Hca-F cells remained between 80% and 100%. Hca-F cells metastasized only to the lymph node, and did not disseminate to other organs of C3H/Hej mice. The percentages of metastasis in inoculated mice and the average percentages of lymph node metastasis in each mouse increased in an almost proportional fashion with the number of implanted cells (r = 0.832, for the percentages of metastasis; r= 0.949, for the average percentages of lymph node metastasis) and time after the implantation (r= 0.933, for the percentages of metastasis; r = 0.959, for the average percentages of lymph node metastasis). This transplantable lymph node metastatic model of mouse hepatocellular carcinoma Hca-F cells in C3H/Hej mice provides a useful tool for the study of the mechanism of tumor lymph node metastasis and may provide a new insight into the development of tumor experimental therapy. PMID- 10367940 TI - Frequent mutations in the beta-catenin gene in desmoid tumors from patients without familial adenomatous polyposis. AB - Mutations in the APC gene contribute to development of sporadic desmoid tumors as well as to the hereditary tumors that usually accompany familial adenomatous polyposis (FAP). Adenomatous polyposis coli (APC) mutations cause an intracellular accumulation of beta-catenin that results in abnormal signaling in the wnt/wingless pathway. Mutations of the beta-catenin gene itself have also been noted in several types of tumors. In this study we screened the beta-catenin gene in 13 sporadic desmoid tumors for alterations in exon 3, which encodes several serine/threonine residues that are targets for phosphorylation by GSK 3beta. Somatic substitutions at codons 41 (threonine) and 45 (serine) were identified in seven independent tumors, respectively. Although no APC mutations were detected among the remaining six tumors, we found accumulation of beta catenin by Western blotting analysis in one such tumor for which frozen tissues were available. Our results have suggested that possible involvement of beta catenin activation by beta-catenin gene mutation or alteration of other factor(s) can contribute to desmoid tumorigenesis. PMID- 10367939 TI - Inhibitory effects of cytokines on ovarian and endometrial carcinoma cells in vitro with special reference to induction of specific transcriptional regulators. AB - In the present study, we have investigated the effects of interferons-alpha (IFN alpha) and -gamma (IFN-gamma), interleukin-10 (IL-10) and -13 (IL-13), transforming growth factor-beta1 (TGF-beta1), granulocyte-macrophage colony stimulating factor (GM-CSF), and tumor necrosis factor-alpha (TNF-alpha) on cell proliferation and induction of transcription factors AP-1 and NF-kappaB in UM-EC 3 human endometrial adenocarcinoma cells and UT-OC-5 ovarian carcinoma cells in vitro. In addition, cellular DNA was extracted to study if any of these factors is able to induce apoptosis. In UM-EC-3 cell line DNA synthesis was inhibited by GM-CSF, IL-10, IL-13, TGF-beta1, IFN-alpha, and IFN-gamma after 48 and 72 h in culture, whereas TNF-alpha had no significant effect on cell proliferation in any of the experiments. The inhibition of DNA synthesis was similarly observed in UT OC-5 ovarian carcinoma cells by IL-10, TNF-alpha, and IFN-gamma after 48 and 72 h, whereas IFN-alpha had no statistically significant effect. An inhibitory effect of GM-CSF was observed only after 48 h and TGF-beta after 72 h in culture, respectively. Transcription factors AP-1 and NF-kappaB were both constitutively active in UM-EC-3 and UT-OC-5 cells. The binding activity of AP-1 was found to be stimulated by all growth-inhibitory cytokines studied in both cell lines, whereas the specific binding activity of NF-kappaB was affected moderately only by TNF alpha in UT-OC-5 ovarian carcinoma cells. No signs of DNA fragmentation typical of apoptosis were observed in any of these studies. PMID- 10367941 TI - Cytotoxicity, cellular uptake, and cellular biotransformations of oxaliplatin in human colon carcinoma cells. AB - Biotransformation products of platinum anticancer drugs have been suggested to be responsible for drug efficacy and toxicity. This study was designed to determine whether the efficacy of the closely related 1,2-diaminocyclohexane-Pt (dach-Pt) compounds oxaliplatin and ormaplatin were determined primarily by the parent drugs or by one of their biotransformation products. Based on consideration of both in vitro cytotoxicity in human colon carcinoma cells (HT-29) and concentrations following oxaliplatin administration in vivo, our data suggest that the efficacy of oxaliplatin is primarily determined by the plasma levels of the parent drug, with the biotransformation products Pt(dach)Cl2, Pt(dach)(H2O)Cl, and Pt(dach)(H2O)2 making only minor contributions. The stable biotransformation products containing amino acids did not have any significant cytotoxicity. In contrast, our data suggest that the efficacy of ormaplatin is primarily determined by plasma levels of Pt(dach)Cl2. The cytotoxicity of oxaliplatin, Pt(dach)Cl2, and Pt(dach)(H2O)Cl was approximately proportional to their cellular uptake, whereas the cytotoxicity of ormaplatin, Pt(dach)(H2O)2, and Pt(dach)(Met) was less than predicted from their uptake. Treatment of HT-29 cells with equimolar external concentrations of Pt(dach)Cl2 and oxaliplatin resulted in the formation of twofold more Pt-DNA adducts following Pt(dach)Cl2 treatment than following oxaliplatin treatment. However, intracellular Pt(dach)Cl2 levels were 30-fold higher for Pt(dach)Cl2-treated cells than for oxaliplatin-treated cells. These data suggest that intracellular conversion of oxaliplatin to Pt(dach)Cl2 makes only a minor contribution to Pt-DNA adduct formation and the resultant cytotoxicity of oxaliplatin. PMID- 10367942 TI - Hypothermia causes a reversible, p53-mediated cell cycle arrest in cultured fibroblasts. AB - Normal human fibroblasts grown in cell culture undergo a reversible growth arrest when incubated at 28 degrees C. During incubation at 28 degrees C, levels of p53 and p21 rise in these cells and cell cycle analysis shows that they have undergone a cell cycle arrest. To examine the importance of p53 in mediating this arrest, mouse embryo fibroblasts that are either wild-type or that are defective in p53 were also subjected to hypothermia. Only those cells with wild-type p53 undergo a cell cycle arrest, indicating that p53 has a role in mediating this response. Because many tumor cells have defective p53, this suggests that hypothermia may increase the selective toxicity of chemotherapeutic agents for tumor cells. PMID- 10367943 TI - Vaccines and their transfection potency. AB - Viral vaccines have been shown to contain residual host cell-DNA. There is no doubt about uptake and expression of foreign DNA in mammalian cells but the mechanism of transfection is not completely understood. It is suggested that DNA associates with several compounds and is transferred into the cell by endocytosis. In this study we estimate the potential of transfection of several original adjuvants by adding reporter plasmid DNA (pDNA) to vaccines. We used fibrosarcoma cells as an in vitro model and the results indicate that the cells are not able to express pDNA. Therefore we propose that adjuvants included in viral vaccines have no potential to transfect fibroblasts. PMID- 10367944 TI - Instruments for oral disease-intervention strategies: recombinant Lactobacillus casei expressing tetanus toxin fragment C for vaccination or myelin proteins for oral tolerance induction in multiple sclerosis. AB - Lactobacillus strains possess properties that make them attractive candidates as vehicles for oral administration of therapeutics. In this report we describe the construction and analysis of recombinant Lactobacillus casei applicable in oral vaccination against an infectious disease (tetanus) and in oral tolerance induction for intervention in an autoimmune disease, multiple sclerosis. Recombinant L. casei which express surface-anchored tetanus toxin fragment C (TTFC) were generated. Quantitative analysis by flow cytometry demonstrated a high level of cell wall-bound expression of TTFC and immunogenicity was demonstrated by parenteral immunization with whole cell extracts of the recombinants. A series of expression vectors was constructed to secrete human myelin basic protein (hMBP) or hMBP as a fusion protein with beta-glucuronidase from Escherichia coli. These heterologous products produced by L. casei were detected in the growth medium and parenteral immunization with this medium evoked antibodies against hMBP, confirming that secretion indeed had occurred. Based on the different localization of the heterologous proteins, lactobacilli expressing surface-anchored TTFC are ideally suited for the induction of antibody responses, whereas lactobacilli that secrete myelin proteins can be used for the induction of peripheral T-cell tolerance. In conclusion, the specific technology described here allows the construction of a wide array of safe live recombinant lactobacilli which may prove to be useful in oral intervention strategies for the prevention of infectious diseases or treatment of autoimmune diseases. PMID- 10367945 TI - Oral vaccination of mice with human papillomavirus virus-like particles induces systemic virus-neutralizing antibodies. AB - To assess whether oral vaccination against human papillomavirus (HPV) may be feasible, we administered HPV virus-like particles (VLPs) to mice by gavage. Enzyme-linked immunosorbent assay (ELISA) results indicated that serum anti-VLP immunoglobulin G (IgG) and IgA antibodies were induced after oral vaccination, and these responses demonstrated antigenic specificities that were conformationally dependent and restricted according to HPV genotype. Importantly, orally induced postimmune sera were found to neutralize HPV-11 virions in vitro. These results indicated that the VLPs were antigenically stable in the environment of the gastrointestinal tract and were able to engage in potentially useful immune system interactions. These findings support the concept of oral vaccination against anogenital HPV disease, and suggest the possibility that this may be a useful approach to the immunization of large populations against cervical cancer and other HPV associated diseases. PMID- 10367946 TI - DNA-based non-invasive vaccination onto the skin. AB - Non-invasive vaccination onto the skin (NIVS) could improve vaccination programs because the procedure requires no specially trained personnel and may eliminate many problems associated with needle injections. There is also evidence that the efficacy of a skin-targeted vaccine may be optimal when the antigen is expressed within the outer layer that is in constant contact with potential pathogens. We report here that non-invasive gene delivery by pipetting adenovirus- or liposome complexed plasmid DNA onto the outer layer of skin could achieve localized transgene expression within a restricted subset of skin in mice and the elicitation of an immune response against the protein encoded by the DNA. These results provide a proof of principle that NIVS may appear as a novel method for the administration of DNA-based vaccines. PMID- 10367947 TI - Protection of calves against cryptosporidiosis with immune bovine colostrum induced by a Cryptosporidium parvum recombinant protein. AB - The purpose of the study was to determine if immunization with a recombinant protein (rC7) of Cryptosporidium parvum would induce immune bovine colostrum that protected calves against cryptosporidiosis following oral challenge with C. parvum oocysts. Late gestation Holstein cows with low titers of antibody to the p23 antigen of C. parvum were immunized three times with 300 microg affinity purified rC7 C. parvum recombinant protein (immune cows), or left nonimmunized (control cows). Colostrum was obtained from each cow in both groups and partitioned into identical aliquots of pooled immune colostrum or pooled control colostrum. Twelve calves obtained at birth received either immune or control colostrum within the first 2 h, and again at 12 and 24 h of age. Each calf was challenged orally with 10(7) C. parvum oocysts at 12 h of age and monitored for signs of cryptosporidiosis. All six calves administered pooled control colostrum developed severe diarrhea (mean total fecal volume = 8447+/-5600 ml) and shed an average of 1.87+/-1.66 x 10(12) C. parvum oocysts. None of the six calves administered pooled immune colostrum developed diarrhea (mean total fecal volume = 740+/-750 ml, p < 0.05), and shed significantly fewer oocysts (3.05+/-2.26 x 10(9), p < 0.05). The absence of diarrhea and 2.79 log10 (99.8%) reduction in oocyst excretion indicates that immune bovine colostrum induced by immunization with C. parvum recombinant protein rC7 provided substantial protection against cryptosporidiosis in neonatal calves. PMID- 10367948 TI - High efficiency gene replacement in Salmonella enteritidis: chimeric fimbrins containing a T-cell epitope from Leishmania major. AB - A simple, high frequency chromosomal gene replacement method of general utility was developed for Salmonella enteritidis. This system uses an unstable, imperfectly segregating, temperature-sensitive replicon, pHSG415, as a carrier of the recombinant gene of interest. It also allows for site-specific replacement of chromosomal genes without the need for antibiotic resistance markers in the recombinant genes or the use of specific bacterial strains. This strategy was used to replace the chromosomal sefA and agfA fimbrin genes of S. enteritidis 3b with recombinant genes containing a 48 bp DNA fragment encoding PT3, an immunoprotective T-cell epitope from GP63 of Leishmania major. The fidelity of chimeric fimbrial replacements were confirmed by DNA sequence analysis. Nearly 30% of the S. enteritidis clones selected in the final stage of sefA mutagenesis contained the sefA::PT3 recombinant gene, whereas for agfA the efficiency was as high as 10%. To our knowledge, this is the first report of fimbrial epitope replacement in the Salmonellae and the first chimeric fimbrin genes that have been reconstituted into a wild-type genetic background for any organism. As such, this model represents a promising 'organelle' expression system for epitope display in vaccinology. PMID- 10367949 TI - Booster vaccination with recombinant hepatitis B vaccine four years after priming with one single dose. AB - We here studied the antibody response to a booster dose four years after the administration of one single dose of recombinant HB vaccine. Before receiving the booster dose, levels of protective antibodies (anti-HBs) were generally low and 24/41 (59%) individuals lacked detectable antibodies (< 1 IU/L). Within 14 d of booster vaccination, 36/38 (95%) vaccinees showed levels of antibodies > 100 IU/L. Notably, these levels were at least as high as those of a reference group 12 months after initiation of vaccination according to the standard three-dose vaccination at intervals of 0, 1 and 6 months. In conclusion, one single dose of HB vaccine seemed to confer on young healthy individuals a well preserved B cell memory, disclosed as a rapid and strong antibody response to a second dose four years later. PMID- 10367950 TI - Gene gun DNA vaccination with Rev-independent synthetic HIV-1 gp160 envelope gene using mammalian codons. AB - DNA immunization with HIV envelope plasmids induce only moderate levels of specific antibodies which may in part be due to limitations in expression influenced by a species-specific and biased HIV codon usage. We compared antibody levels, Th1/Th2 type and CTL responses induced by synthetic genes encoding membrane bound gp160 versus secreted gp120 using optimized codons and the efficient gene gun immunization method. The in vitro expression of syn.gp160 as gp120 + gp41 was Rev independent and much higher than a classical wt.gp160 plasmid. Mice immunized with syn.gp160 and wt.gp160 generated low and inconsistent ELISA antibody titres whereas the secreted gp120 consistently induced faster seroconversion and higher antibody titres. Due to a higher C + G content the numbers of putative CpG immune (Th1) stimulatory motifs were highest in the synthetic gp160 gene. However, both synthetic genes induced an equally strong and more pronounced Th2 response with higher IgG1/IgG2a and IFNgamma/IL-4 ratios than the wt.gp160 gene. As for induction of CTL, synthetic genes induced a somewhat earlier response but did not offer any advantage over wild type genes at a later time point. Thus, optimizing codon usage has the advantage of rendering the structural HIV genes Rev independent. For induction of antibodies the level of expression, while important, seems less critical than optimal contact with antigen presenting cells at locations reached by the secreted gp120 protein. A proposed Th1 adjuvant effect of the higher numbers of CpG motifs in the synthetic genes was not seen using gene gun immunization which may be due to the low amount of DNA used. PMID- 10367951 TI - Specific IgG to gelatin in children with systemic immediate- and nonimmediate type reactions to measles, mumps and rubella vaccines. AB - We examined anti-gelatin IgG in sera of children who suffered from systemic adverse reactions upon immunization with gelatin-containing live virus vaccines. In the group of 30 children who had immediate-type reactions and anti-gelatin IgE, 30 (100%) had anti-gelatin IgG and 29 (96%) had anti-gelatin IgG4. In another group of 75 children who had nonimmediate-type reactions and no anti gelatin IgE, 22 (29%) had anti-gelatin IgG and six (8%) had IgG4. The IgG positivity well correlated with the lymphocyte proliferation assay positivity. In contrast, as a negative control, all 24 children who had no allergic reaction to live virus vaccines had no anti-gelatin IgG and IgG4. The results suggest that immune-response to gelatin may play a role in the pathogenesis of systemic nonimmediate-type reactions to the live virus vaccines. PMID- 10367952 TI - Comparative immunogenicity and tolerance of Vaqta and Havrix. AB - In an open-label, randomised trial, 520 adults of both sexes aged 18-30 years were allocated to receive one of two inactivated hepatitis A vaccines; Vaqta or Havrix, at 0 and 24 weeks. Doses used were 50 or 100 antigen units (U) of Vaqta and 1440 enzyme linked immunosorbent assay U of Havrix given as 1 ml intramuscular injections. For each trial group safety data were available for all subjects and full serological data for more than 80% of randomised volunteers. Local side effects which were mild in most cases were significantly (p < 0.0001) more common with Havrix than with Vaqta, irrespective of dose given. Systemic tolerance was similar for the 3 regimens. From 4 weeks after the first dose, > or =94% of the subjects had seroconverted. The mean antibody titres 4 weeks after the second vaccine dose were 2978, 4346 and 1589 mIU/ml in subjects who were randomised to Vaqta 50 U/dose, Vaqta 100 U/dose and Havrix 1440 U/dose, respectively. The 2 vaccines had similar immunogenicity but local tolerance was better with Vaqta. PMID- 10367953 TI - Development of recombinant ovalbumin-luteinizing hormone releasing hormone as a potential sterilization vaccine. AB - The objective was to develop an immunogenic chimeric ovalbumin-LHRH (ova-LHRH) molecule using genetic engineering. Hybrid ova-LHRH genes with either four or seven LHRH inserts were constructed by cassette mutagenesis and oligonucleotide mismatch mutagenesis. Recombinant ova-LHRH proteins were over-expressed in E. coli strain BL21 (DE3) using a pET expression system, which expresses a target protein with a C-terminal His-Tag. The C-terminal His-Tag allows purification by metal chelation chromatography. The antigenicity and biological effects of these recombinant proteins were tested in mice. In experiment 1, 17 female 7 wk old BALB/c mice were randomly divided into three groups. Six mice were injected with 50 microg of the recombinant ovalbumin (ova) protein. Five mice were injected with 50 microg of the recombinant protein with four LHRH inserts (ova-LHRH-7). Six mice were injected with 50 microg of the recombinant protein with seven LHRH inserts (ova-LHRH-7). One primary immunization using Freund's complete adjuvant was followed by one booster using incomplete adjuvant. Mice were killed 2 wk after the booster, blood collected, and the reproductive tract removed and weighed. Only ova-LHRH-7 decreased (P < 0.01) uterine-ovarian weight (89+/-11 mg) vs control (138+/-6 mg) and ova-LHRH-4 (126+/-16 mg). The genetically engineered molecule with seven LHRH inserts induced LHRH antibody titers which were significantly correlated (r = -0.79) with biological response. In experiment 2, the recombinant ova-LHRH-7 was evaluated at two doses with the adjuvants Zmax and Immumax. Seventy female 6-8 wk old BALB/c mice were randomly divided into seven groups of 10 mice each. Anti-LHRH titers were detected in all of the ova-LHRH-7 immunized mice. Significant decreases were shown in uterine-ovarian weight of the mice by the immunization with 30 microg of ova-LHRH-7 and Zmax (P < 0.005) or 10 microg of ova-LHRH-7 with Immumax (P < 0.025). These data show that the recombinant ova-LHRH-7 protein could have potential as an effective sterilization vaccine. PMID- 10367954 TI - Cationic lipid DC-Chol induces an improved and balanced immunity able to overcome the unresponsiveness to the hepatitis B vaccine. AB - Th1 and Th2 immune responses against antigens can be modulated by the use of adjuvants. Since antibody isotypes (IgG1 and IgG2a) and cytokines induced may reflect the Th differentiation taking place during the immune response, the humoral and cellular immune responses induced in mice against hepatitis B virus surface antigen (HBsAg) were examined when the antigen was either adsorbed to aluminum hydroxyde or administered with a new adjuvant the cationic lipid 3beta [N-(N',N'-dimethylaminoethane)carbamoyl]cholesterol (DC-Chol). The use of DC-Chol increased antibody responses in responding BALB/c mice, induced more consistent IgG1 and IgG2a antibody responses in OF1 mice and overcame the nonresponse to HBsAg in B10.M mice. Furthermore, DC-Chol was able to induce cellular immune responses to HBsAg. The DC-Chol induced a balanced Th1/Th2 response, which enabled mice to overcome the inherited unresponsiveness to HBsAg encountered with aluminum-adjuvanted vaccine. Thus, the DC-Chol provides a signal to switch on both Th1 and Th2 responses, which may have important implications for vaccination against hepatitis B virus, as well as for enhancing weak immunogenicity of other recombinant purified antigens in a nonresponder population. PMID- 10367955 TI - UV responses in the retina of the turtle. AB - To study processing of UV stimuli in the retina of the turtle, Trachemys dorbignii, we recorded intracellular responses to spectral light from 89 cells: 54 horizontal (47 monophasic, five (R/G) biphasic and two (Y/B) triphasic), 14 bipolar, 12 amacrine, and nine ganglion cells. Spectral sensitivities were measured with monochromatic flashes or with the dynamic constant response method in dark or chromatic adapted states. Stray light and second-order harmonics were also measured. (1) All cells responded to UV stimuli, although none had maximum sensitivity in the UV. (2) Most horizontal, bipolar, and amacrine cells had red peaked spectral sensitivities. (3) Red adaptation of all monophasic horizontal cells indicated a single red input, except one that had additional peaks in the blue and UV. (4) Responses of biphasic and triphasic horizontal cells to UV light were always hyperpolarizing. Opposition between hyperpolarizing and depolarizing responses at long wavelengths indicates that UV responses were not due to the beta band of red receptors. (5) An unstained spectrally opponent bipolar cell hyperpolarized in the center to green light and antagonistically depolarized in the surround to UV, blue, and green flashes, but hyperpolarized to red. (6) All dark-adapted amacrine cells were red-peaked monophasic cells, but red adaptation broadened their spectral-sensitivity curves or displaced their peaks. An A15, an A18, and an A24 wide-field amacrine cell were stained. (7) A G15 bistratified ganglion cell is shown here for the first time to be spectrally opponent. This UVB/RG cell depolarized to UV and blue and hyperpolarized to red and green. It differs from previously reported turtle ganglion cells in being color opponent in the entire field, not only in the surround, and in showing spatial opponency. PMID- 10367956 TI - Responses of directionally selective retinal ganglion cells to activation of AMPA glutamate receptors. AB - Previous studies in the rabbit retina have shown that drugs which block AMPA glutamate receptors abolish directional selectivity in ON-OFF directionally selective (DS) ganglion cells. The effects of activation of AMPA receptors on the directionally selective responses of these ganglion cells had not been studied. In the present study, extracellular recordings of the responses of ON-OFF DS ganglion cells to a moving bar of light were made in an in vitro rabbit retinal preparation. In control solution, bath application of AMPA (7-10 microM) abolished the light responses of most ON-OFF DS ganglion cells. On washout of AMPA, the light responses rapidly returned; however, the cells temporarily lost the ability to discriminate the direction of the moving bar of light. That is, the cells responded equally to movement in the preferred and null directions. Pretreatment of retinas with the glycine receptor antagonist strychnine (1-2 microM) did not alter the effects of AMPA. On the other hand, in retinas pretreated with the GABA(A) receptor antagonist SR95531 (0.2-0.25 microM), AMPA did not abolish the light responses of ON-OFF DS ganglion cells but instead abolished directional selectivity in these cells by bringing out a response to movement in the null direction. This finding suggests that an AMPA-induced GABA efflux from cells in the retina was responsible for the suppression of the light responses by AMPA. In control solution, application of the selective AMPA receptor agonist (S)-5-fluorowillardiine (2-3 microM) only temporarily abolished the light responses of ON-OFF DS ganglion cells. As the light responses returned, it was clear that directional selectivity had been abolished by (S)-5 fluorowillardiine. In control solution, blocking AMPA receptor desensitization with cyclothiazide (80-100 microM) greatly reduced the light responses of ON-OFF DS ganglion cells. As the light responses slowly returned on washout of cyclothiazide, directional selectivity was clearly reduced although not abolished. In retinas pretreated with SR95531, application of cyclothiazide abolished directional selectivity. Diazoxide (700-1000 microM), another blocker of AMPA receptor desensitization, abolished directional selectivity in ON-OFF DS ganglion cells without the need of adding SR95531 to the bathing solution. It is concluded that, in the rabbit retina, AMPA receptors play an important role in generating directional selectivity in ON-OFF DS ganglion cells. Moreover, excessive activation of AMPA receptors greatly compromises the mechanism for directional selectivity in ON-OFF DS ganglion cells. PMID- 10367957 TI - 5-HT2a receptors in the rabbit retina: potential presynaptic modulators. AB - Three 5-HT receptors have been implicated in retinal processing but positive identification of the receptors and the localization of receptor subtypes in the retina have not been achieved. In this study, molecular techniques were used to identify one class of 5-HT receptor--5-HT2a--in the retina, and immunohistochemical techniques were used to localize the receptor in the retinal network. Reverse transcription polymerase chain reaction (RT-PCR) techniques were used to identify a segment of the rabbit 5-HT2a gene; a 422 base fragment was identified, cloned, and sequenced. The fragment shows a high degree (ca. 90%) of nucleotide sequence identity with the 5-HT2a receptor gene from other mammals. 5 HT2a immunoreactivity was seen in both the inner and outer plexiform (synaptic) layers of the retina. Using cell-type-specific markers, the 5-HT2a immunoreactivity was shown to be on the terminals of photoreceptor and rod bipolar cells. This association of 5-HT2a receptors with these two synapses suggests that serotonin may be a modulator of synaptic function in the retina. PMID- 10367959 TI - Visual function in regenerating teleost retina following cytotoxic lesioning. AB - Teleost fish retinas can regenerate in vivo in adulthood. Retinal and visual function was assessed in adult goldfish following comprehensive retinal destruction by intraocular injection of ouabain. Electroretinograms (ERGs) and the dorsal light reflex (DLR) were used to evaluate the return of visual function. ERGs were detectable in regenerating eyes 50 to 70 days following ouabain injection. Amplitudes of both a- and b-waves increased steadily through day 210 following ouabain treatment, at which time a-wave amplitude was 90% and b wave amplitude approached 50% of the contralateral control eye. The progressive gain observed in the a-wave was attributed to photoreceptor regeneration. The increase in b-wave amplitude was attributed to an increase in the number of inner nuclear layer cells and the number and efficacy of neuronal connections to or within the inner retina. The photopic spectral sensitivity of the b-wave in regenerating retina closely matched the intrafish control retina, suggesting that the relative numbers of cone photoreceptors was normal in regeneration. The recovery of the DLR (indicated by improved postural balance during regeneration) paralleled electrophysiological gains during retinal regeneration. Fish displayed a marked longitudinal body imbalance toward the control eye following retinal destruction. Improvement in equilibrium was correlated with increasing b-wave amplitudes. When the b-wave reached 50% of control amplitude (30 weeks), normal posture was restored. The return of the ERG indicates that photoreceptors and their synaptic connections must be functional in regenerating retina. Failure of the retina to regenerate produced an abnormal DLR that persisted through 30 weeks and ERGs were not measurable. The return of normal equilibrium indicates that the regenerating retina can establish central connections to the brain, and that the regenerated connections can mediate functional visual behavior. PMID- 10367958 TI - Distribution of the glycine transporter glyt-1 in mammalian and nonmammalian retinae. AB - We have examined the distribution of the glycine transporter glyt-1 in retinae of macaques, cats, rabbits, rats, and chickens. In all species, all glycine containing amacrine cells expressed immunoreactivity for glyt-1, though the intensity of immunoreactivity for glyt-1 did not appear to directly correlate with the intensity of immunoreactivity for glycine in individual cells. A small subpopulation of glycine-immunoreactive displaced amacrine cells or ganglion cells also expressed glyt-1 in retinae from macaques, cats, chickens, and rats but not in retinae from rabbits. In addition, in all species examined, some displaced amacrine cells also contained glycine but did not express glyt-1. In monkeys, cats, and rats, populations of cells which we interpret as being glycine containing interplexiform cells expressed glyt-1: these cells lacked a content of glutamate, suggesting they are not bipolar cells. The glycine-containing bipolar cells did not express glyt-1, suggesting that these cells probably acquired their content of glycine by other means such as via gap junctional connections with glycine-containing amacrine cells. PMID- 10367960 TI - Analysis of dendritic arbors of native and regenerated ganglion cells in the goldfish retina. AB - The retinas of adult teleost fish can regenerate following injury, but little is known about the neuronal integration of the visual scene that is performed by the regenerated retina. Using goldfish retinal ganglion cells (RGCs) as the experimental system, an evaluation of dendritic arbor structure and passive electrotonic properties was developed, the aim being to quantitatively test the hypothesis that native and regenerated RGC dendritic arbors have similar structural and modeled electrotonic attributes. Fractal dimension was chosen as the descriptor of RGC dendritic arbor complexity, and the arbors' transfer function magnitudes were estimated using an electrically passive, equivalent circuit analysis. For both native and regenerated RGCs, arbors qualitatively judged to be simple tended to have lower fractal dimension values than arbors judged to be more complex. All cells had similar cut-off frequencies, and for random stimulation of greater than 25% of an RGC's population of dendritic tips, there was a positive correlation between fractal dimension and transfer function magnitude. Some regenerated RGCs had abnormally long primary dendrites, but neither the distributions of fractal dimension values, nor the estimated transfer function magnitudes, were significantly different between native and regenerated RGCs. The results appear to support the hypothesis that structural and modeled electrotonic attributes of regenerated goldfish RGCs are similar to those of native RGCs, suggesting that regenerated RGCs may restore normal visual function. PMID- 10367961 TI - Excitatory amino acid-induced inositol phosphate formation in cultured retinal pigment epithelium. AB - Excitatory amino acid (EAA)-induced production of inositolphosphates (IPs) was studied in primary cultures of chick retinal pigment epithelium (RPE) following in vitro incorporation of [3H] myo-inositol. Glutamic acid (L-glu) significantly increased [3H]-IPs accumulation (215%). L-glu agonists stimulated [3H]IPs accumulation in the following order of efficiency: N-methyl-D-aspartate (NMDA) > or = L-glu > quisqualate > or = kainate > (IS,3R)-1-aminocyclopentane-1,3 dicarboxylic acid (ACPD). Stimulation was dependent on external Ca2+. The NMDA induced response was blocked by (+)-5-methyl-10,11-dihydro-5H-dibenzo-cyclohepten 5,10-imine maleate (MK-801) and 3-(2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid (CPP) and was decreased by the L-Ca2+-channel blockers verapamil and nifedipine as well as by dantrolene. The metabotropic glutamate receptor (mGluR) antagonist (+)-alpha-methyl-4-carboxyphenylglycine (+)MCPG inhibited 3,5 dihydroxyphenylglycine (DHPG) and ACPD-induced stimulation, which demonstrates the presence in RPE of mGluRs 1 and/or 5, as well as NMDA receptors coupled directly, or through the influx of external Ca2+, to phospholipase C activation. L-glu agonists showed no effect either on basal level of intracellular cyclic adenosine monophosphate, nor on forskolin- or carbachol-induced stimulation of adenylyl cyclase. Since L-glu is released from the retina upon illumination, and receptors for this compound are present in RPE, the activation of the inositide pathway could be involved in the regulation of retina-RPE interaction, which is essential for the visual process. PMID- 10367962 TI - Origin of tectal cholinergic projections in amphibians: a combined study of choline acetyltransferase immunohistochemistry and retrograde transport of dextran amines. AB - Immunohistochemistry for choline acetyltransferase (ChAT) revealed an extensive network of cholinergic fibers in the tectum of amphibians. The distribution of ChAT immunoreactive fibers was not restricted to superficial retinocipient layers, but also included deep tectal layers. The aim of the present study was to investigate the origin of the cholinergic inputs to the tectum of amphibians. For that purpose, application of retrograde tracers in the tectum of the anuran Rana perezi and the urodele Pleurodeles waltl was combined with ChAT immunohistochemistry. Double-labeled cells were found primarily in the nucleus isthmi of both species. The cholinergic isthmotectal projection is bilateral and topographically arranged and all retrogradely labeled cells found in this nucleus were ChAT immunoreactive. Remarkably, abundant cholinergic cells in two tegmental nuclei, the pedunculopontine tegmental nucleus (anurans) and the laterodorsal tegmental nucleus (anurans and urodeles), were demonstrated to provide additional cholinergic innervation to the tectum. We compare the present results with previously reported studies in amphibians and other vertebrates, and discuss the possible functional significance of the cholinergic innervation of the amphibian tectum. PMID- 10367963 TI - TTX attenuates surround inhibition in rabbit retinal ganglion cells. AB - Patch-clamp recordings were made from ganglion cells in an in vitro dark-adapted rabbit retina preparation. Cells were stimulated by images generated on a computer monitor and focussed onto the photoreceptors. Excitatory inward currents were recorded in response to spot stimuli centered on the somas of the recorded cells. Center illumination of on-brisk-transient cells produced large transient excitatory postsynaptic currents (EPSCs) which were invariably followed by a small steady-state inward component. Illumination of a centered annulus failed to elicit the transient EPSC. Simultaneous illumination of the annulus and the center spot blocked the large transient EPSC, consistent with activation of an inhibitory surround. Application of tetrodotoxin (TTX), which blocks sodium dependent action potentials, also blocked the surround inhibition in ON-brisk transient cells as well as some other classes of ganglion cells. It is concluded that, in some ganglion cell classes, the surround is generated largely through the activity of spiking neurons, and it is suggested that the amacrine cells in the inner plexiform layer are involved. PMID- 10367964 TI - Distribution of photoreceptor types in the retina of a marsupial, the tammar wallaby (Macropus eugenii). AB - Mammalian retinae generally contain low numbers of short-wavelength-sensitive cones (S-cones) and higher numbers of middle- to long-wavelength-sensitive cones (M-cones). Some recent studies found topographic differences between the different photoreceptor types and in some instances between photoreceptors and ganglion cells. To investigate this question further, we constructed topographical maps of the different photoreceptors found in an Australian marsupial, the tammar wallaby. We used two polyclonal antibodies that have been shown to label S-cones (JH455) or M-cones (JH492) in a range of mammals. In the tammar wallaby, the antisera clearly distinguish two cone types. JH455 recognizes a small subset of cones (S-cones) with a density of less than 500 cells/mm2 in the ventral retina. Their density increases towards the dorsal retina to about 1600-2000 cells/mm2. JH492 recognizes all remaining cones (M-cones), but also faintly labels most cone cells recognized by JH455. The distribution of M-cones, unlike that of the S-cones, shows a clear horizontal streak of high cell density through the central retina, just like the ganglion cells. Unlike the ganglion cells, however, the M-cones do not peak in the temporal retina but show a very broad peak (12,000-18,000 cells/mm2) in the central or even slightly nasal retina. Based on our findings, the retina of the tammar can be divided into three distinct regions: firstly, the dorsal retina, which has a low ganglion and low cone cell density but a high percentage of S-cones (30%), is thought to provide good spectral sensitivity; secondly, the central horizontal band of retina, which has a high ganglion and high cone cell density and therefore provides good spatial resolution; and thirdly, the ventral retina, which has a low ganglion cell but high cone cell density with few S-cones (5%) and is therefore thought to have a high contrast sensitivity but low acuity. PMID- 10367965 TI - A model for the intracortical origin of orientation preference and tuning in macaque striate cortex. AB - We report results of numerical simulations for a model of generation of orientation selectivity in macaque striate cortex. In contrast to previous models, where the initial orientation bias is generated by convergent geniculate input to simple cells and subsequently sharpened by lateral circuits, our approach is based on anisotropic intracortical excitatory connections which provide both the initial orientation bias and its subsequent amplification. Our study shows that the emerging response properties are similar to the response properties that are observed experimentally, hence the hypothesis of an intracortical generation of orientation bias is a sensible alternative to the notion of an afferent bias by convergent geniculocortical projection patterns. In contrast to models based on an afferent orientation bias, however, the "intracortical hypothesis" predicts that orientation tuning gradually evolves from an initially nonoriented response and a complete loss of orientation tuning when the recurrent excitation is blocked, but new experiments must be designed to unambiguously decide between both hypotheses. PMID- 10367966 TI - Balanced interactions in ganglion-cell receptive fields. AB - Receptive fields of retinal ganglion cells in turtle have excitatory and inhibitory components that are balanced along the dimensions of wavelength, functional ON and OFF responses, and spatial assignments of center and surround. These components were analyzed by spectral light adaptations and by the glutamate agonist, 2-amino-4-phosphonobutyric acid (APB). Extracellular recordings to stationary and moving spots of light were used to map changes in receptive fields. ON spike counts minus OFF spike counts, derived from flashed stationary light spots, quantified functional shifts by calculating normalized mean response modulations. The data show that receptive fields are not static, but rather are dynamic arrangements which depend on linked, antagonistic balances among the three dimensions of wavelength, ON and OFF response functions, and center/surround areas. PMID- 10367967 TI - Ganglion cells of a short-wavelength-sensitive cone pathway in New World monkeys: morphology and physiology. AB - We have studied the morphology and physiology of retinal ganglion cells of a short-wavelength-sensitive cone (SWS-cone) pathway in dichromatic and trichromatic New World anthropoids, the capuchin monkey (Cebus apella) and tufted ear marmoset (Callithrix jacchus). In Old World anthropoids, in which males and females are both trichromats, blue-ON/yellow-OFF retinal ganglion cells have excitatory SWS-cone and inhibitory middle- and long-wavelength-sensitive (MWS- and LWS-) cone inputs, and have been anatomically identified as small-field bistratified ganglion cells (SB-cells) (Dacey & Lee, 1994). Among retinal ganglion cells of New World monkeys, we find SB-cells which have very similar morphology to such cells in macaque and human; for example, the inner dendritic tree is larger and denser than the outer dendritic tree. We also find blue-on retinal ganglion cells of the capuchin to have physiological responses strongly resembling such cells of the macaque monkey retina; for example, responses were more sustained, with a gentler low frequency roll-off than MC-cells, and no evidence of contrast gain control. There was no difference between dichromatic and trichromatic individuals. The results support the view that SWS-cone pathways are similarly organized in New and Old World primates, consistent with the hypothesis that these pathways form a phylogenetically ancient color system. PMID- 10367968 TI - Effects of inhibiting glutamine synthetase and blocking glutamate uptake on b wave generation in the isolated rat retina. AB - The purpose of the present experiments was to evaluate the contribution of the glutamate-glutamine cycle in retinal glial (Muller) cells to photoreceptor cell synaptic transmission. Dark-adapted isolated rat retinas were superfused with oxygenated bicarbonate-buffered media. Recordings were made of the b-wave of the electroretinogram as a measure of light-induced photoreceptor to ON-bipolar neuron transmission. L-methionine sulfoximine (1-10 mM) was added to superfusion media to inhibit glutamine synthetase, a Muller cell specific enzyme, by more than 99% within 5-10 min, thereby disrupting the conversion of glutamate to glutamine in the Muller cells. Threo-hydroxyaspartic acid and D-aspartate were used to block glutamate transporters. The amplitude of the b-wave was well maintained for 1-2 h provided 0.25 mM glutamate or 0.25 mM glutamine was included in the media. Without exogenous glutamate or glutamine the amplitude of the b wave declined by about 70% within 1 h. Inhibition of glutamate transporters led to a rapid (2-5 min) reversible loss of the b-wave in the presence and absence of the amino acids. In contrast, inhibition of glutamine synthetase did not alter significantly either the amplitude of the b-wave in the presence of glutamate or glutamine or the rate of decline of the b-wave found in the absence of these amino acids. Excellent recovery of the b-wave was found when 0.25 mM glutamate was resupplied to L-methionine sulfoximine-treated retinas. The results suggest that in the isolated rat retina uptake of released glutamate into photoreceptors plays a more important role in transmitter recycling than does uptake of glutamate into Muller cells and its subsequent conversion to glutamine. PMID- 10367969 TI - The dynamics of primate M retinal ganglion cells. AB - The retinal ganglion cells (RGCs) of the primate form at least two classes--M and P--that differ fundamentally in their functional properties. M cells have temporal-frequency response characteristics distinct from P cells (Benardete et al., 1992; Lee et al., 1994). In this paper, we elaborate on the temporal frequency responses of M cells and focus in detail on the contrast gain control (Shapley & Victor, 1979a,b). Earlier data showed that the temporal-frequency response of M cells is altered by the level of stimulus contrast (Benardete et al., 1992). Higher contrast shifts the peak of the frequency-response curve to higher temporal frequency and produces a phase advance. In this paper, by fitting the data to a linear filter model, the effect of contrast on the temporal frequency response is subsumed into a change in a single parameter in the model. Furthermore, the model fits are used to predict the response of M cells to steps of contrast, and these predictions demonstrate the dynamic effect of contrast on the M cells' response. We also present new data concerning the spatial organization of the contrast gain control in the primate and show that the signal that controls the contrast gain must come from a broadly distributed network of small subunits in the surround of the M-cell receptive field. PMID- 10367970 TI - Current source-density analysis of light-evoked field potentials in rabbit retina. AB - The technique of current source-density analysis was applied to several components of the light-evoked field potentials (electroretinogram) from the retina of the superfused eyecup of rabbit. The depth distributions of the major current sources and sinks were: b-wave--sink at outer plexiform layer, source at inner plexiform layer; M-wave--sink at inner plexiform layer, source at retinal surface; and slow PIII--source near outer plexiform layer, sink at retinal surface. These distributions, along with the sensitivities of these responses to certain pharmacological agents, support earlier studies that Muller cells generate the M-wave and slow PIII, but that depolarizing bipolar cells directly generate the b-wave. PMID- 10367971 TI - Light and electron microscopical analysis of nitric oxide synthase-like immunoreactive neurons in the rat retina. AB - We have investigated the morphology of the NOS-like immunoreactive neurons and their synaptic connectivity in the rat retina by immunocytochemistry using antisera against nitric oxide synthase (NOS). In the present study, several types of amacrine cells were labeled with anti-NOS antisera. Type 1 cells had large somata located in the inner nuclear layer (INL) with long and sparsely branched processes ramifying mainly in stratum 3 of the inner plexiform layer (IPL). Somata of type 2 cells with smaller diameters were also located in the INL. Their fine processes branched mostly in stratum 3 of the IPL. A third population showing NOS-like immunoreactivity was a class of displaced amacrine cells in the ganglion cell layer (GCL). Their soma size was similar to that of the type 1 cells; however, their processes stratified mainly in strata 4 and 5 of the IPL. Labeled neurons were evenly distributed throughout the retina, and the mean densities were 57.0 +/- 9.7 cells/mm2 for the type 1 cells, 239.3 +/- 43.4 cells/mm2 for the type 2 cells and 121.2 +/- 27.5 cells/mm2 cells for the displaced amacrine cells. The synaptic connectivity of NOS-like immunoreactive amacrine cells was identified in the IPL by electron microscopy. NOS-labeled amacrine cell processes received synaptic input from other amacrine cell processes and bipolar cell axon terminals in all strata of the IPL. The most frequent postsynaptic targets of NOS-immunoreactive amacrine cells were other amacrine cell processes. Ganglion cell dendrites were also postsynaptic to NOS like immunoreactive neurons in both sublaminae of the IPL. Synaptic outputs onto bipolar cells were observed in sublamina b of the IPL. In addition, a few synaptic contacts between labeled cell processes were observed. Our results suggest that NOS immunoreactive cells may be modulated by other amacrine cells and ON cone bipolar cells, and act preferentially on other amacrine cells. PMID- 10367973 TI - Diversity of temporal self-organized behaviors in a biochemical system. AB - The numerical study of a glycolytic model formed by a system of three delay differential equations revealed a notable richness of temporal structures which included the three main routes to chaos, as well as a multiplicity of stable coexisting states. The Feigenbaum, intermitency and quasiperiodicity routes to chaos can emerge in the biochemical oscillator. Moreover, different types of birhythmicity, trirhythmicity and hard excitation emerge in the phase space. For a single range of the control parameter it can be observed the coexistence of two quasiperiodicity routes to chaos, the coexistence of a stable steady state with a stable torus, and the coexistence of a strange attractor with different stable regimes such as chaos with different periodic regimes, chaos with bursting behavior, and chaos with torus. In most of the numerical studies, the biochemical oscillator has been considered under periodic input flux being the mean input flux rate 6 mM/h. On the other hand, several investigators have observed quasiperiodic time patterns and chaotic oscillations by monitoring the fluorescence of NADH in glycolyzing yeast under sinusoidal glucose input flux. Our numerical results match well with these experimental studies. PMID- 10367972 TI - The electroretinogram of the rhodopsin knockout mouse. AB - The electroretinogram (ERG) of the rhodopsin knockout (rho-/-) mouse of Humphries et al. (1997) (Humphries et al., 1997) was studied for evidence of light-evoked rod activity and to describe the cone function. The rho-/- retina develops normal numbers of rod and cone nuclei, but the rods have no outer segments, and no rhodopsin is found by immunohistochemistry. The dark-adapted ERG threshold was elevated 4.7 log units above wild-type (WT) control mice, indicating that any residual rod responses were reduced >50,000-fold, consistent with a complete functional knockout. The dark-adapted rho-/- ERG had a cone waveform, and the spectral sensitivity peaked near 510 nm for both dark-adapted and light-adapted conditions, without evidence of a Purkinje shift. The light-adapted ERG b-wave amplitude of young rho-/- mice was the same as WT. The amplitude remained steady up to postnatal day P47, but thereafter it declined to only 1-2% by P80 when no cone outer segments remained. Cone b-wave threshold of dark-adapted rho-/- mice was -1.07 +/- 0.39 log cd-s/m2 (n = 17), which is 1.27 log units more sensitive than light-adapted thresholds against a rod-suppressing Ganzfeld background of 1.61 log scotopic cd/m2. This indicates that dark-adapted WT responses to still dimmer stimuli are exclusively rod driven with minimal cone intrusion. Above this cone threshold intensity, the dark-adapted b-wave of WT will be a summation of rod and cone responses. Threshold versus intensity (TVI) studies gave no evidence of a rod influence on the mouse cone b-wave. PMID- 10367974 TI - Transient phase of enzyme reactions. Time course equations of the strict and the rapid equilibrium conditions and their computerized derivation. AB - In this contribution, we present the derivation, from the strict transient phase equations of enzyme reactions, of the transient phase equations under the usual assumptions that one or more of the reversible steps involved in the mechanism of the enzyme reaction are assumed to be in rapid equilibrium. Moreover, we present the transient phase equations of all of the species in a general enzyme system model, valid for the partial or total rapid equilibrium conditions, as well as the particular case of the strict transient phase equations. In the case of the rapid equilibrium assumptions, the equations may be given either as functions of the individual rate constants in the reversible steps assumed in rapid equilibrium or as functions of the corresponding equilibrium constants. The steady state equations are easily obtained from the transient phase equations by setting the time --> infinity. We have implemented a computer program, easy to use and with a user-friendly format for the input of data and output of results, which allows the user to derive the symbolic strict transient phase equations and/or those corresponding to the assumption that one or more of the reversible reaction steps are in rapid equilibrium. PMID- 10367975 TI - Life after Newton: an ecological metaphysic. AB - Ecology may indeed be 'deep', as some have maintained, but perhaps much of the mystery surrounding it owes more simply to the dissonance between ecological notions and the fundamentals of the modern synthesis. Comparison of the axioms supporting the Newtonian world view with those underlying the organicist and stochastic metaphors that motivate much of ecosystems science reveals strong disagreements--especially regarding the nature of the causes of events and the scalar domains over which these causes can operate. The late Karl Popper held that the causal closure forced by our mechanical perspective on nature frustrates our attempts to achieve an 'evolutionary theory of knowledge.' He suggested that the Newtonian concept of 'force' must be generalized to encompass the contingencies that arise in evolutionary processes. His reformulation of force as 'propensity' leads quite naturally to a generalization of Newton's laws for ecology. The revised tenets appear, however, to exhibit more scope and allow for change to arise from within a system. Although Newton's laws survive (albeit in altered form) within a coalescing ecological metaphysic, the axioms that Enlightenment thinkers appended to Newton's work seem ill-suited for ecology and perhaps should yield to a new and coherent set of assumptions on how to view the processes of nature. PMID- 10367976 TI - The emergence of the concept of a tool in food-retrieving behavior of the ants Formica japonica Motschulsky. AB - We propose a weak definition for the usage of a tool for an ethological study of ants. In particular, we illustrate the usage of a cart in experiments on the transportation of foods by ants as employing a logical structure including a contradiction. The contradiction originates in ruling out the very term 'tool' from the description of the behavior of the animals. Focusing on a self-similar structure underlying the description of a contradiction, we observe a particular time-series sequence of ants' behaviors following a 1/f or Zipf's law. The behaviors following the 1/f or Zipf's law manifest an appropriateness of the notion of a cart as a logical jump. PMID- 10367977 TI - Chronological changes in the ganglioside composition of human milk during lactation. AB - The normal chronological changes in the ganglioside composition of human milk during lactation were examined by means of a high-performance thin-layer chromatography (HPTLC) micro-method with 1 ml of milk from each lactation. Six human milk ganglioside compositions were found, which were designated as GM3, GD3, GX1, GX2, GX3 and GX4. GX1-GX4, which had not been described previously, were tentatively assumed to be gangliosides of the c-series because they did not react to the GA1 antibody after sialidase treatment. GD3 was the major composition of the colostrum (GD3, 42-56%; GM3, 2.22-6.5%). GM3 increased sharply at eight days postpartum (GD3, 32.22%; GM3, 27.79%) and then increased gradually after eight days until examined at seven weeks postpartum (GM3/GD3, 0.84-2.67). The newly found GX1-GX4 showed some variability in the percentage composition between individuals, and there were no distinct differences between the colostrum and the later milk. The drastic compositional changes in GM3 and GD3 during lactation might have some biological significance, such as in immunological activity, somatic growth and the nervous system. PMID- 10367978 TI - Health status of a population of infants born before 26 weeks gestation derived from routine data collected between 21 and 27 months post-delivery. AB - This retrospective study was designed: (a) to determine the extent to which routine data sources in the UK can provide data relating to the later health status of selected groups of infants; and (b) to use such an approach to describe the outcome of a geographically defined population of infants born before 26 weeks gestation. All infants of less than 26 weeks gestation admitted for neonatal intensive care during the period 1/1/91 and 31/12/93 whose mother's address at the time of birth was within the boundaries of the Trent Health Region were included. Health status was assessed against a previously described simple scheme and using information from existing sources only. During the 3-year period 249 infants of less than 26 weeks gestation were admitted for intensive care. Of these 66 (26.5%) survived to be discharged from the neonatal service. A further seven infants died before the age of 2 years. Of the remaining 59 four were lost to follow up (three could not be traced; one was living abroad). Of the 55 infants reviewed, 36 demonstrated no features, pre-defined in the classification scheme, of severe disability. However, only 30 children appeared to be considered entirely normal. CONCLUSION: Infants born before 26 weeks gestation and admitted for neonatal intensive care had, approximately, a 12% chance of normal survival to 2 years. A slightly smaller proportion of infants survived with significant disability. Existing routine data sources could be adapted to provide useful public health information about the outcome of 'high risk' groups of infants. PMID- 10367979 TI - Laminin alpha2 deficient congenital muscular dystrophy: prenatal diagnosis. AB - Laminin alpha2 chain-deficient congenital muscular dystrophy (CMD) is diagnosed by genetic analysis and by immunohistochemistry. Since laminin alpha2 chain is expressed in placental trophoblasts, the demonstration of its deficiency in chorionic villi is a useful aid to prenatal diagnosis. We present our experience with the use of the immunohistochemical method for prenatal diagnosis in four women, all of whom had at least one child with laminin alpha2 chain-deficient CMD. Immunohistochemistry provided a rapid procedure for prenatal diagnosis, and follow-up of these four cases confirmed its reliability. PMID- 10367980 TI - Maturation of body and breathing movements in 24-33 week-old fetuses threatening to deliver prematurely. AB - Maturation of spontaneous fetal body and breathing movements of 24- to 33-week old fetuses in 168 pregnancies threatening to deliver prematurely were examined on the basis of newborn outcome (premature compromised, premature healthy, term healthy). Maturation of fetuses in 60 low-risk pregnancies delivering as healthy full-term infants served as a normative comparison group. Each fetus was observed for 30 min; the amount of body and breathing movements were noted and an estimation of amniotic fluid volume was made. The pattern of behavioural maturation was similar for all outcome groups; with advancing gestation there was a decrease in body movements and an increase in breathing movements. Both reduced activity levels and advanced behaviours were observed in the high-risk outcome groups. The high-risk fetuses had reduced levels of body movements which increased with better outcome and, an earlier onset of increased amounts of breathing, occurring at 30 weeks in contrast to 33 weeks for the comparison group. In the presence of ruptured membranes, those high-risk fetuses who were born prematurely had less breathing compared to those who delivered at term. Similar maturation patterns among high- and low-risk outcome groups suggests normal/typical functional development in the high-risk fetal groups. The observed differential behaviours were associated with prematurity and most likely associated with events leading to premature labour. PMID- 10367981 TI - The neurodevelopmental outcome of term infants with different intrauterine growth characteristics. AB - The objective of this study was to test the hypothesis that neurodevelopmental outcome would differ between two groups of small-for-gestational age infants born at term showing different in utero growth characteristics during the third trimester. The design was a prospective cohort study done at a tertiary referral centre. The 76 subjects who fulfilled the inclusion criteria had an estimated fetal weight below the 10th centile for their gestation. Subsequent to enrolment, repeat ultrasound scans were performed weekly to determine growth velocity. Twenty-three infants whose change in fetal abdominal circumference between first and last scan was greater than -1.5 standard deviation scores (SDS) were assigned to the intrauterine growth retardation group (IUGR) while 53 infants whose fetal abdominal circumference changed less than 1.5 SDS were assigned to the small-for gestational-age (SGA) group. Ten infants with normal intrauterine growth were enrolled as controls. Following delivery all infants had a neurological examination and a cranial ultrasound scan. At 1 year, 75 infants (87%) were traced and reassessed (49 SGA, 18 IUGR and eight controls) with a neurological examination and a developmental assessment. At birth, impairments were found in 27 (51%) of the SGA, 13 (57%) of the IUGR groups and one (10%) of the controls. At 1 year, 18 (37%) of the SGA subjects, six (33%) of the IUGR subjects and one (13%) of the control infant were impaired, including three (6%) of the SGA subjects and one (6%) of the IUGR subjects who were disabled. We conclude that in term fetuses with an estimated birthweight below the 10th centile for their gestation, the pattern of growth in the third trimester does not affect outcome at 1 year. In spite of optimum obstetric management, nearly one-third of the combined SGA and IUGR term fetuses had suffered some, albeit minor, neurological damage. PMID- 10367982 TI - The effect of sleep state on electroretinographic (ERG) activity during early human development. AB - To assess the effects of sleep state on human retinal electric responses, full field electroretinograms were obtained in a cross-sectional study from 123 preterm infants at 36, 40 and 56 weeks of post-gestational age. At each age, electroretinographic recordings were assigned to one of two groups according to whether the infants were in active sleep or quiet sleep. Both sleep states were determined behaviorally. Pure rod, maximal, 30 Hz flicker and light adapted single cone responses were evaluated when a sleep state was clearly established. Peak-to-peak amplitudes of most electroretinographic responses were significantly larger in active sleep relative to quiet sleep at 36 and 40 weeks of post gestational age. We speculate that larger amplitudes during active sleep may play a role in the maturation of the visual system. PMID- 10367983 TI - The effect of early feeding on the neonatal blood glucose level at 1-hour of age. AB - Early infant feeding after birth is being promoted, although it is unclear whether this has any effect on carbohydrate metabolism. We planned to measure the capillary glucose at one hour (1 h) of age in a group of infants from non diabetic pregnancies using the HemoCue B-Glucose system to see if the timing and method of early feeding would influence this result. Seventy-five term infants were studied, 22 of which were breast-fed, 24 bottle fed and 29 not fed during the first hour after birth. The mean whole blood glucose results were 2.34 mmol/l, 2.52 mmol/l and 2.58 mmol/l respectively (P = NS). The first two groups were fed at a median of 22 minutes before sampling. We conclude that the timing and method of early feeding in the newborn have no significant effect on the blood glucose level measured at 1 h of age, and this remains a reliable outcome measure in studies of glucose metabolism in pregnancy. PMID- 10367984 TI - Three-dimensional ultrasonographic imaging of fetal tooth buds for characterization of facial clefts. AB - The purpose of this prospective study was to investigate whether the antenatal characterization of fetal facial clefts can be improved by three-dimensional ultrasonographic visualization of fetal tooth buds. Between January 1996 and June 1998, seventeen consecutive fetuses with facial clefts were examined for fetal maxillary tooth buds in the cleft area using three-dimensional multiplanar reconstruction. It was possible in all cases to classify the clefts either as cleft lip alone or unilateral cleft lip and palate or bilateral cleft lip and palate. Three-dimensional computed tomography and histological jaw sections of three stillborn infants were produced in order to examine the correlation between the sonographic, radiographical and histological findings. The prenatal characterization of the facial clefts by means of a visualization of the tooth buds showed to be accurate postnatally in all cases. The sonographic proof of tooth buds might gain increasing importance as this technique seems to facilitate and improve the prenatal classification of suspected facial clefts. PMID- 10367985 TI - Implications of a neural network model of early sensori-motor development for the field of developmental neurology. AB - This paper reports on a neural network model for early sensori-motor development and on the possible implications of this research for our understanding and, eventually, treatment of motor disorders like cerebral palsy. We recapitulate the results we published in detail in a series of papers [1-4]. The neural circuits in the model self-organize on the basis of rhythmic activity spontaneously generated in the model. This indicates the importance of endogenously generated activity in the developing brain. We also show that afferent feed-back from the mechanical part of the model is easily incorporated in the neural part of the model. In this way the model acquires reflex-related properties which have long been demonstrated in man. In the discussion we relate these experimental findings to the variability concept from developmental neurology and show how variable motor performance is important for motor learning. We also discuss possible implications of our modelling effort for movement disorders, specifically spastic cerebral palsy. PMID- 10367987 TI - Motor activity and perception of sleep in depressed patients. AB - Subjectively experienced sleep patterns often differ from observed sleeping behavior in insomniacs. Sleep patterns have been evaluated by measurements of motor activity in healthy subjects and insomniacs, but results may depend on the insomnia subtype. Sleep disturbances are frequent complaints in depression, but the influence of psychopathology on activity measurements remains elusive. Therefore, the relationship between reported sleep complaints and motor activity was studied in patients with major depression. Severity of depression was documented in depressed inpatients by observer-(HAMD) and self-rated scales (DACL, SHAPS-D). Self-reports of sleep were obtained by Pittsburgh Sleep Quality Index (PSQI) and daily sleep logs (DSL). Motor activity was continuously recorded over 72 h by actigraphy. 'Good' sleepers showed less motor activity during the night compared to 'poor' sleepers (p<0.01). Patients with high HAMD scores (> or = 18) showed greater nocturnal motor activity and less sleep quality compared to patients with low HAMD scores (p<0.01). When controlling for age and severity of depression, partial correlation was found to be significant between perceived daily sleep quality and nocturnal motor activity (r = -0.63, p<0.01). There was a significant effect of nocturnal motor activity as a covariate on disturbances of subjective sleep quality and severity of depression as the main effect (p<0.01). In depressed patients, nocturnal motor activity seems to be an indicator of experienced sleep disturbances. The results warrant further controlled studies to evaluate the role of psychological factors for objective measurements and subjective perception of sleep patterns. PMID- 10367986 TI - The rationale for corticotropin-releasing hormone receptor (CRH-R) antagonists to treat depression and anxiety. AB - Neuroendocrine studies strongly suggest that dysregulation of the hypothalamic pituitary-adrenocortical (HPA) system plays a causal role in the development and course of depression. Whereas the initial mechanism resulting in HPA hyperdrive remains to be elucidated, evidence has emerged that corticosteroid receptor function is impaired in many patients with depression and in many healthy individuals at increased genetic risk for an depressive disorder. Assuming such impaired receptor function, then central secretion of CRH would be enhanced in many brain areas, which would account for a variety of depressive symptoms. As shown in rats and also in transgenic mice with impaired glucocorticoid receptor function, antidepressants enhance the signaling through corticosteroid receptors. This mechanism of action can be amplified through blocking central mechanisms that drive the HPA system. Animal experiments using antisense oligodeoxynucleotides directed against the mRNA of both CRH receptor subtypes identified the CRH1 receptor as the mediator of the anxiogenic effects of CRH. Studies in mouse mutants in which this receptor subtype had been deleted extended these findings as the animals were less anxious than wild-type mice when experimentally stressed. Thus, patients with clinical conditions that are causally related to HPA hyperactivity may profit from treatment with a CRH1 receptor antagonist. PMID- 10367988 TI - Galanin has REM-sleep deprivation-like effects on the sleep EEG in healthy young men. AB - Rapid eye movement (REM) sleep deprivation leads to an induction of galanin gene expression in the rat brain, especially in the hypothalamus. Galanin affects neuroendocrine systems that are involved in sleep regulation, i.e. the growth hormone-releasing hormone-dependent system of the hypothalamus and the locus coeruleus. In the study reported here we investigated the effects of 4 x 50 microg galanin (n = 10) and of 4 x 150 microg galanin (n = 8) administered hourly between 22.00 and 01.00 h as intravenous boluses on the sleep EEG and nocturnal hormone secretion in healthy young men. Galanin administration significantly increased REM sleep in the third sleep cycle with no difference between the two doses. Spectral analysis revealed a significant increase in the EEG power in the delta and theta frequency range for the total night after the lower dose of galanin, but not after the higher dose. The secretion of growth hormone, cortisol and prolactin remained unchanged during sleep in both cases. Our data are consistent with the assumption of a functional resemblance between the effect of galanin and that of REM sleep deprivation, which is known to have antidepressive efficacy. PMID- 10367989 TI - Course of treatment received by depressed patients. AB - Using data from an observational study of affective disorders, we describe the rates of transition among levels of antidepressant treatment for subjects with Major Depressive Disorder (MDD), and relate these changes to changes in clinical status. We report on the treatment received during the first 10 years of follow up in the Collaborative Depression Study by 555 patients with a diagnosis of MDD of at least one month's duration. This work extends the initial examination of treatment received during the first eight weeks after entry into this study that showed depressed patients to be on low levels of treatment. Multiplicative intensity models which generalize survival analysis models were used to analyse these data. Description of the course of treatment of these depressed patients shows that low levels of treatment persist for these patients across subsequent episodes, and that these episodes, like the index one, are characterized by extended time in a symptomatic subcriterion state after acute symptoms have improved. These long-term descriptions of treatment support the initial hypothesis that these CDS patients were undertreated. The long-term tendency toward undertreatment seems to persist even as newer treatments become available and widely accepted in practice. PMID- 10367990 TI - Gender differences in depression: an ethological study of nonverbal behavior during interviews. AB - Previous studies of gender differences in the phenomenology of depression have focused mostly on symptoms as measured by self-report questionnaires or clinician rated scales. In this study, we examined gender differences in the interpersonal behavior of depressed patients by using ethological techniques which involve direct observation of behavior. The nonverbal behavior of 72 nondepressed volunteers and 68 patients with a DSM-III-R diagnosis of nonpsychotic unipolar depression was videorecorded during clinical interviews and scored according to an ethological scoring system including 37 behavior patterns, mostly facial expressions and hand movements. Both male and female depressed patients showed a global restriction of nonverbal expressiveness reflecting a tendency towards social withdrawal. Nonverbal expression of hostility was the only behavioral category on which depressed patients scored higher than nondepressed volunteers. Even though clinical status exerted marked effects on the ethological profile, depression did not obscure some important differences in the nonverbal behavior of males and females. As a group, depressed women showed more socially interactive behaviors than depressed men. Their modality of interacting included higher levels both of nonverbal hostility and of submissive and affiliative behaviors. These results are discussed in view of clinical data indicating a relationship between gender, style of social interaction and response to antidepressant drugs. PMID- 10367992 TI - Medial frontal cortex involvement in PTSD symptoms: a SPECT study. AB - The regional cerebral blood flow (rCBF) responses to a combat stress-related auditory stimulus was examined in Vietnam veterans diagnosed with posttraumatic stress disorder (PTSD). Based on prior data in healthy subjects, we hypothesized that the medial prefrontal cortex may be involved in the processing of stress responses. Twelve male veterans diagnosed with PTSD, 11 age-matched, combat exposed subjects without PTSD, and 12 healthy control subjects were studied with single-photon emission tomography and the blood flow tracer [99mTc]-HMPAO. Subjects were studied twice, while listening to combat sounds or white noise. Significant increases in the blood flow to the medial prefrontal cortex were observed in PTSD patients, but not in the control groups, which correlated at trend levels with psychophysical measures of stress response. These data support the involvement of the medial prefrontal cortex in the pathophysiology of PTSD, possibly mediating some of its symptoms. PMID- 10367991 TI - Dissociation between I2-imidazoline receptors and MAO-B activity in platelets of patients with Alzheimer's type dementia. AB - The I2-imidazoline receptor is expressed in brain and platelets and could represent a new binding domain on MAO-B enzyme. Brain I2-imidazoline receptors and MAO-B sites have been found to be increased in Alzheimer's disease. The study sought to evaluate I2-imidazoline receptors and MAO-B activity in platelets from patients with Alzheimer's type dementia (ATD) and matched controls. Preliminary saturation experiments of [3H]idazoxan binding to platelet purified mitochondrial membranes were performed to determine the maximal number of binding sites (Bmax) and the apparent dissociation constant (Kd). Afterwards, the I2-imidazoline receptor density ([3H]idazoxan at 8 and 20 nM in the presence of 2 x 10(-6) M efaroxan) was evaluated in 20 patients with ATD and 17 controls. MAO-B activity was quantified by [14C]PEA oxidation. All subjects were screened for cognitive evaluation by the Mini-Mental State Examination. The density of I2-imidazoline receptors was similar in ATD patients (8.4 and 14.3 fmol/mg protein) and controls (8.3 and 14.0 fmol/mg protein). MAO-B activity was 22% higher in ATD subjects. Significant correlations between I2-imidazoline receptors and MAO-B activity were observed. No relationships between I2-imidazoline receptors or MAO-B activity and the cognitive score were observed. In conclusion, platelet I2-imidazoline receptors do not show the increase of I2-imidazoline receptors previously observed in brain of subjects with ATD. The dissociation between I2-imidazoline receptors and MAO-B in platelets suggests that the enzyme contributes to but not exclusively represents the I2-imidazoline receptor. PMID- 10367993 TI - Phenotypic characteristics of Obsessive-Compulsive Disorder ascertained in adulthood. AB - Over the past decade, the increased awareness and knowledge of Obsessive Compulsive Disorder (OCD) has allowed the in-depth study of its phenotypic characteristics. The largest studies to date have described the symptom and syndrome characteristics of treatment-seeking patients. While usefully homogeneous with regard to their current state, the clinical characteristics of patients seeking treatment may only partially represent the OCD population. We report findings from 100 self-selected volunteers at various stages of their OCD illness who were participating in a genetic study. Many similarities with past reports were found, including high rates of mood disorder, significantly more mood disorder in females as compared with males, and increased social impairment among males despite an equal amount of time in episodes of disorder. On the other hand, mean age of onset in this nontreatment seeking population was younger. Lifetime rates of obsessions and compulsions in this population were substantially higher than previous reports, suggesting that the content of obsessions and compulsions shifted over time, and evolved into a lifetime repertoire. Furthermore, a separate analysis of the age of clinically significant O-C symptom onset without impairment revealed that males and females did not differ, suggesting that previous reports of earlier onset age in males may actually reflect earlier onset of impairment. Future genetic studies may benefit from the analysis of both significant O-C symptom onset, as well as the onset of full-syndromal OCD. These findings may suggest phenotypic characteristics that define homogeneous subgroups of patients with OCD. PMID- 10367994 TI - Trait versus state aspects of the MMPI during the early course of schizophrenia. AB - Scores on the Minnesota Multiphasic Personality Inventory (MMPI)-168 item version were examined during periods of clinical remission and of psychosis for recent onset schizophrenia patients (n = 19) and at comparable time intervals for demographically matched normal participants (n = 19). To determine diagnostic specificity, MMPIs for participants with bipolar affective disorder in remission (n = 12) were also examined. Methods for distinguishing between stable vulnerability indicators, mediating vulnerability factors and episode indicators of psychopathology were adapted from Nuechterlein and Dawson (1984). MMPI scales Pa, Sc and validity scale F showed a combination of trait and state qualities, characteristic of mediating vulnerability factors. These scales reflect changes that occur during psychotic episodes but also apparently tap personality characteristics that endure into periods of clinical remission. Unexpectedly, some MMPI scales that are not typically associated with psychotic disorders (i.e. Hs, D, and Hy) were significantly higher in schizophrenia patients across psychotic and clinically remitted states than in normal participants. In clinical remission, higher scores on scales Hs, D and Hy, showed some specificity to schizophrenia relative to bipolar disorder. While MMPI-168 scales Pd and Pt fit the pattern for vulnerability indicators, it was uncertain whether they belonged to the 'stable' versus 'mediating' subtype. MMPI scores that continue to be higher in remission than in a normal sample may reflect either enduring vulnerability factors or the impact of schizophrenia and the individuals' attempts to cope with the disorder. Studies of first-degree relatives will be needed to provide converging evidence that certain personality characteristics reflect genetic predisposition to schizophrenia. PMID- 10367996 TI - Exercise and the regulation of energy intake. AB - Energy balance is the resultant of ingested calories and energy expenditure and is generally maintained within narrow limits over prolonged periods. Exercise leads to an increase in energy expenditure which is, in the long-term, counteracted by increased energy intake. Evidence for this comes from a study in voluntarily running female rats that increased their daily food intake to 130% of the sedentary controls. In contrast, when considered on a short-term basis, exercise will suppress food intake to prevent a potentially dangerous disruption of energy substrate homeostasis. Studies in permanently cannulated rats submitted to a test meal and 2 hrs swimming reveal that both food intake and exercise lead to increases in glucose and free fatty acid (FFA) levels in the blood. These changes in glucose and FFA, combined with the exercise-induced alteration in among others glucagon, corticotropin releasing hormone (CRH) and body temperature, may lead to the short-term anorexic effect of exercise. PMID- 10367995 TI - Tower of Hanoi and WCST performance in schizophrenia: problem-solving capacity and clinical correlates. AB - We administered a computerized version of WCST, a well established test, sensitive to executive function deficits in schizophrenia that involves many features of cognitive processing, and of Tower of Hanoi, a test that may offer cognitive challenges more specifically related to planning and sequencing, to 28 schizophrenic patients and 28 matched controls to examine a worthwhile question regarding the relative ability of these two tasks to differentiate schizophrenia and normal groups as well as exploring the relationship of these two instruments to clinical variables. The schizophrenic patients performed significantly worse than normal subjects both on Tower of Hanoi test and on WCST. The discriminant analysis identified in a multivariate way a pattern of indexes that differentiate the two groups. This pattern, characterized by specific indexes of WCST and TOH, could suggest the existence of a common underlying factor that determines the cognitive impairment in problem-solving of schizophrenics. These findings and the relationship with positive and negative symptoms have been discussed in the light of the model of the impairment in the internal representation of context information. PMID- 10367997 TI - Skeletal muscle substrate metabolism. AB - Endurance power of muscles is determined largely by the capacities to oxidize substrates in mitochondria in the process of making ATP by oxidative phosphorylation. This review explores physiological and morphological factors that may cause limitation of carbohydrate and fat utilization by muscle cells. The pathways for oxygen and substrates converge in muscle mitochondria. In mammals, a structural limitation of carbohydrate and lipid transfer from the microvascular system to muscle cells is reached at a moderate work intensity (that is, at 40-50% of VO2max). At higher work rates intracellular substrate stores must be used for oxidation. Because of the importance of these intracellular stores for aerobic work we find larger intramyocellular substrate stores in endurance trained athletes. The transfer limitations for carbohydrates and lipids on the level of the sarcolemma implies that the design of the respiratory cascade from lungs to muscle mitochondria reflects primarily oxygen demand. PMID- 10367998 TI - Exercise training and substrate utilisation in obesity. AB - Together with diet and behavioural modification, regular exercise is one of the key components of programs for the treatment of obesity. It appears to be one of the major factors determining the long-term success of weight loss programs. One of the mechanisms that may underlie this and other beneficial effects of regular exercise is the effect of exercise training on substrate utilisation. Exercise training increases fat oxidation in lean subjects. The capacity to mobilise and oxidise fat has been shown to be impaired in obese and post-obese subjects. Thus, an increase in the capacity for fat oxidation may help to maintain fat and energy balance at a lower fat mass in individuals with a predisposition for obesity. However, in view of the impaired fat oxidation in obese and post-obese individuals the question arises whether exercise training also increases fat oxidation in the obese. Few studies have addressed this question and have investigated the effect of exercise training on substrate utilisation in obesity. Addition of an exercise training program has been shown to prevent the reduction of basal fat oxidation that is associated with diet-induced weight loss in two studies. No effect of additional exercise training on substrate oxidation during exercise was found. In post-obese subjects exercise training caused no change in 24 h substrate oxidation in one study and increased carbohydrate, rather than fat oxidation in another. In obese subjects neither low (40% maximal aerobic fitness (VO2max)) nor high (70% VO2max) intensity training was found to affect resting substrate oxidation. During exercise fractional fat oxidation was increased after low, but not after high intensity training. The question, whether exercise training increases fat oxidation in obese and post-obese as in lean subjects therefore cannot be answered conclusively at this point in time and requires further study. The role of exercise intensity and type of exercise needs to be studied as well. PMID- 10367999 TI - Role of physical activity in the prevention of obesity in children. AB - The increasing prevalence of childhood obesity and its concomitant health risks justify widespread efforts toward prevention. Although both diet and physical activity have been emphasized as appropriate interventions, the current paper focuses on the role of physical activity in obesity prevention. Children's levels of physical activity are highly variable, and may be influenced by a multitude of factors including physiological, psychological, sociocultural and environmental determinants. Although the relationship between physical activity and obesity is controversial and the protective mechanism unclear, physical activity is hypothesized to protect individuals from the development of obesity by increasing energy expenditure and resting metabolic rate (RMR) and leading to a favourable fuel utilization. The beneficial effect of physical activity in children is supported by controlled exercise intervention programs. Several broad-based public health interventions designed to increase children's levels of physical activity have been implemented in schools, families and communities, with results suggesting promising strategies for the prevention of childhood obesity. It is likely that successful prevention of childhood obesity through physical activity promotion will involve theory-based, culturally appropriate school, family and community interventions. Through policy changes, environmental planning and educational efforts in schools and communities, increased opportunities and encouragement for physical activity can be provided. PMID- 10368000 TI - Lifestyle and obesity in adolescence and young adulthood: results from the Amsterdam Growth And Health Longitudinal Study (AGAHLS). AB - OBJECTIVE: To investigate if there is a longitudinal relationship between the development of fat mass in males and females (aged between 12-28 y) and their lifestyle, with respect to diet and physical activity. DESIGN: In the Amsterdam Growth And Health Longitudinal Study (AGAHLS), a group of 500 boys and girls are being followed from the age of 13 y (mean age 13.5 y), over a period of 20 y until the age of 32 y (mean age 32.5 y). SUBJECTS AND MEASUREMENTS: Data from AGAHLS are analysed of six repeated measurements of growth (body height, body mass, four skinfolds), health parameters with respect to cardiovascular disease (CVD) (obesity, hypertension, hypercholesterolaemia) and two important lifestyle tractors: physical activity (PA) (weighted energy output) and dietary intake (DI) (total energy intake, contribution of fat, carbohydrate and protein) of about 200 males and females between the ages of 13-27 y. RESULTS: The longitudinal results of PA show a steep decrease in the energy expenditure from the age of 13 y from about 4500 Mets per week to 3000 Mets per week at the age of 27 y in both sexes. The longitudinal results of DI also show a decrease in energy intake per kilogram body mass from about 225 kJ per day at the age of 13 y to about 155 kJ at the age of 27 y. During adolescence, boys show a 15% higher energy intake and a 20% higher energy expenditure than girls. At the age of 27 y, the difference between the sexes in energy intake is reduced to 10% and the difference in energy expenditure disappears. Results of tracking analyses over the period of 15y indicate a low stability coefficient of PA (0.34; 0.19-0.49), but higher coefficients of DI (0.55; 0.45-0.64) and fat mass (0.63; 0.56-0.71). The search for important lifestyle factors that can discriminate high- from low-risk participants for a high fat mass, estimated from the sum of four skinfolds and body mass (BM), resulted in an Odds Ratio (OR) of 1.5 (1.2-1.8) with daily intake of proteins and an OR of 0.81 (0.69-0.96) with PA. Surprisingly the fat mass is negatively related with the daily intake of energy per kg BM (OR: 0.37; 0.28 0.49). The longitudinal relation between fat mass and PA (corrected for DI) between the ages of 13-27 y, showed a significant inverse relationship (P<0.01) if fat mass was estimated from the sum of four skinfolds, but not if estimated from the body mass index (BMI). CONCLUSIONS: Over the adolescent and young adult period from 13-27 y, fat mass indicates a fairly good predictability. A high PA in both sexes is related with a low fat mass. Therefore promotion of habitual physical activity in the adolescent period seems effective in the early prevention of obesity. PMID- 10368001 TI - Physical activity and the prevention of childhood obesity--Europe versus the United States. PMID- 10368002 TI - Physical activity assessment with accelerometers. AB - OBJECTIVE: Evaluation of motion sensors, specifically accelerometers, as an objective tool for the assessment of physical activity in large populations, over periods long enough to be representative of normal daily life and with minimal discomfort to the subjects. METHOD: Review of validation studies of accelerometers with indirect calorimetry as a reference method. Accelerometers were commercially available one-axial accelerometers: Caltrac, Computer Science Application (CSA) accelerometer, Mini Motionlogger Actigraph; the tri-axial accelerometer Tritrac-R3 D; and an tri-axial accelerometer for movement registration (Tracmor) from our laboratory. RESULTS: There is no clear difference for correspondence between indirect calorimetry and accelerometer counts during level walking, whether one-axial or tri-axial and placed at the wrist, hip or low back. Sedentary activities are better reflected with a tri-axial accelerometer than with a one-axial accelerometer. Two accelerometers were validated in free living conditions with doubly labeled water. The highest correlation between accelerometer output and activity induced energy expenditure was found for Tracmor. CONCLUSIONS: From all accelerometers tested, the tri-axial accelerometer for movement registration is an objective method that can be used to distinguish differences in activity levels between individuals and assess the effect of interventions on physical activity within individuals. PMID- 10368003 TI - Physical activity and weight maintenance. AB - Physical activity is an important component of a weight-reducing program. When combined with a low fat diet, a physical activity program can reduce body weight by 10-15% in individuals complying with the program. However, even in disciplined individuals, resistance to lose fat ultimately occurs generally before the body composition status of the reduced-obese subjects is comparable to that of their lean counterparts. On the other hand, this weight loss is generally sufficient to normalize the risk profile regarding the development of diabetes and heart diseases. Therefore, this suggests that trying to lose weight beyond the threshold of spontaneous resistance to lose fat, may be unnecessary and not feasible. Furthermore, if one also considers the potential risks for health associated with large fat loss, it is probably not relevant to encourage further weight loss in reduced-obese individuals, when their metabolic profile is normalized. It is also important to emphasize that in such a context, the physical activity program must be maintained on a permanent basis to prevent body weight regain. PMID- 10368004 TI - Genetic determinants of sports participation and daily physical activity. AB - OBJECTIVE: The purpose is to review the existing literature on genetic determinants of sports participation, daily physical activity (PA) resting metabolic rate (RMR) and activity as a temperamental trait. DESIGN: A synthesis will be given of the published material on this topic with special focus on twin and family data, and association and linkage studies. MEASUREMENTS: Self reported sports participation, daily PA, RMR and activity as a temperamental trait. ANALYSIS: Transmission and heritability coefficients calculated from twin and family data will be reported. RESULTS: The reported heritability coefficients for sports participation vary between 0.35-0.83, and those for daily PA between 0.29 0.62. If one of the parents or co-twins is active in sports, it is more likely that the child or co-twin is also active in sports (odds ratios (ORs) vary from 1.2-5.8). Twin and parent-child correlations for RMR also indicate a moderate genetic effect. At present, only a linkage between RMR and uncoupling protein 2 markers has been demonstrated. CONCLUSION: The genetic determination of sports participation, daily PA and RMR, varies from low to moderately high, and only between the uncoupling protein 2 genetic marker and RMR has a linkage has been demonstrated. PMID- 10368005 TI - Physical activity, obesity and blood lipids. AB - PURPOSE: To present the evidence concerning the influence of physical activity on the dyslipidaemia of obesity and overweight. METHODS: Review of a personal library of literature on the interactions of physical activity, lipoprotein metabolism and body fatness. SUMMARY OF FINDINGS: Obesity, in particular abdominal obesity, is associated with dyslipidaemia--specifically elevated plasma concentrations of triacylglycerol (TAG) in the fasted state, an exaggerated postprandial rise in plasma TAG, low concentrations of high density lipoprotein cholesterol (HDL) and possibly a preponderance of small dense low density lipoproteins. Regular physical activity contributes to the avoidance of overweight and hence to the development of dyslipidaemia. Although low levels of body fatness contribute to the high levels of HDL cholesterol and the low levels of TAG in trained people there are other important determinants of these characteristics. In particular, exercise (and probably training, that is regular, frequent exercise over months and years) enhances the metabolic capacity for TAG, possibly through mechanisms involving increased activity of lipoprotein lipase. This, in turn, has effects on other lipoprotein species such that the transport of TAG and cholesterol in the circulation is improved. There is evidence for a dose-response relationship, with for example, higher levels of HDL cholesterol in men and women who expend more energy in exercise. For the majority of healthy, sedentary adults frequent, moderate intensity exercise equivalent to a total gross energy expenditure of about 8.5 MJ per week is probably a sufficient to influence lipoprotein lipids. PMID- 10368006 TI - An evaluation of epoxy resin phantom materials for megavoltage photon dosimetry. AB - Epoxy resin phantom materials have been available for some time and are widely used for dosimetry purposes, not least in audit phantoms. Information on their behaviour is partly available in the literature, but there are different mixes and formulations often given similar names and it may not be appropriate to transfer information from one material to another. Five commercially available water substitute materials have been evaluated for use in megavoltage photon beams: WT1, WTe, RMI 451, RMI 457 and 'plastic water'. Four independent experiments were carried out to compare these materials with water in megavoltage photon beams ranging in energy from cobalt 60 to nominal 16 MV x-rays, and some general conclusions are drawn from the results as to their use. All are suitable for relative dosimetry in megavoltage photon beams. However, differences of up to 1% are observed for absolute measurements. The newer formulations, developed for electron beam use, are also closer to water for megavoltage photon beams. PMID- 10368007 TI - Patient dosimetry in abdominal arteriography. AB - This study aims at accurate quantification of x-ray exposure and effective dose to the patient in abdominal arteriography. Using an automatic monitoring system, all relevant exposure parameters were determined during 172 abdominal arteriographies. Common projections were extracted for a 'normal' reference group of procedures and used in Monte Carlo calculations of dose-area product to organ dose conversion coefficients. Dose-area product, organ doses and effective dose were quantified for intravenous and intra-arterial procedures. The large data sets describing exposure could be condensed to a set of 28 common views. New coefficients to convert dose area product to organ equivalent dose and effective dose were calculated for nine views contributing approximately 80% to the total dose-area product. The average dose-area product was 32 Gy cm2 in intravenous procedures and 47 Gy cm2 in intra-arterial procedures. The corresponding average effective doses to the patient were 4 mSv and 6 mSv respectively (range 2-12 mSv, actual value depending on procedure type and gender). It is concluded that automatic monitoring of x ray exposure parameters, complemented by the calculation of Monte Carlo organ dose conversion coefficients, is a feasible and promising approach to accurate dosimetry of complex arteriographic procedures. PMID- 10368008 TI - The magnetization transfer characteristics of human breast tissues: an in vitro NMR study. AB - A series of freshly excised human breast tissues was analysed using a nuclear magnetic resonance spectrometer and then subjected to routine histopathology examination. Tissues comprised normal parenchymal, adipose, fibrocystic, fibroadenoma and malignant types. An inversion-recovery sequence performed both with and without magnetization transfer allowed T1, T1s, M0 and Ms values to be obtained. From this information, the magnetization transfer rate constant, K, was calculated for each tissue sample. These data show that T1s provided greater discrimination between neoplasic and normal tissues than did T1. However, neither T1s nor K values provided a means of discriminating between benign and malignant disease. PMID- 10368009 TI - Evaluation of spurious results in the infrared measurement of CO2 isotope ratios due to spectral effects: a computer simulation study. AB - The application of infrared spectroscopy to the measurement of carbon isotope ratio breath tests is a promising alternative to conventional techniques, offering relative simplicity and lower costs. However, when designing such an instrument one should be conscious of several spectral effects that may be misinterpreted as changes in the isotope concentration and which therefore lead to spurious results. Through a series of computer simulations which model the behaviour of the CO2 absorption spectrum, the risk these effects pose to reliable measurement of 13CO2/12CO2 ratios and the measures required to eliminate them are evaluated. The computer model provides a flexible high-resolution spectrum of the four main isotopomer fundamental transitions and fifteen of their most significant hotband transitions. It is demonstrated that the infrared source, infrared windows and breath sample itself all exhibit strong temperature induced errors but pressure effects do not produce significant errors. We conclude that for reliable measurement of 13CO2/12CO2 ratios using infrared spectroscopy no pressure controls are required. window effects are eliminated using windows wedged at a minimum angle of 0.8-2.2 mrad, depending on the material, and the temperature sensitivity of source and gas cells necessitates stabilization to an accuracy of at least 0.2 K. PMID- 10368010 TI - Laser shaping of corneal transplants in vitro: area ablation with small overlapping laser spots produced by a pulsed scanning laser beam using an optimizing ablation algorithm. AB - Area laser lathing and trephination of donor corneas is used to produce different types of grafts for human transplantation. 193 nm (ArF excimer) laser radiation is used, since this is known to give a non-thermal laser-tissue interaction with a minimal zone of tissue damage. To guarantee the highest degree of flexibility concerning the overall shape of the grafts as well as their thickness profiles, we use a small (compared with the area to be ablated) scanning laser spot. For area lathing of the tissue we have developed a new ablation algorithm (optimized scanning laser ablation, OSLA) that can be applied to lathe and perforate any tissue--with concave (as in this application), convex or plane surface geometry- where surface precision and smoothness are key issues. Using OSLA with the Excimer Laser Corneal Shaping System (a tool for in vitro fabrication of all kinds of corneal transplants like donor buttons for keratoplasty, lamellar grafts for epikeratoplasty and refractive lenticules) enabled us to produce all types of corneal grafts with very high precision. This is considered to be a major improvement towards the production of refractive lenticules. PMID- 10368011 TI - Alanine/EPR dosimetry in brachytherapy. AB - The paper reports the experimental procedure adopted to determine the absorbed dose rate in water per reference air kerma rate, D(Kr), (d, theta), along the transverse bisector axis of a 137Cs brachytherapy source. The dose rate measurements have been carried out at difference distances, d, from the source using alanine dosemeters in a water phantom. The reference air kerma rate, Kr, was determined adopting a 'direct procedure' that uses a spherical ionization chamber in air. The dose rate constant of the source examined was D(Kr) (1, pi/2) = 0.99 +/- 0.03 cGy h(-1) (microGy h(-1))(-1). The values of the radial dose function along the transverse axis, g(d), determined with an uncertainty of 3.4% (1sigma), were found to be in good agreement with the results reported in the literature. The uncertainty in dose rate value has been estimated as 2.8% (1sigma) for distances from the source up to 7 cm. Kr has been determined with 1.2% (1sigma) uncertainty. So D(r) (d, pi/2) values were determined with 3% (1sigma) uncertainty. PMID- 10368012 TI - Determination of water absorbed dose in a carbon ion beam using thimble ionization chambers. AB - The method to measure absorbed dose to water in a field of carbon ions as applied for the heavy ion therapy project at the Heavy Ion Research Laboratory in Darmstadt (GSI), Germany, is described in detail. Thimble ionization chambers with a water absorbed calibration factor are applied. The dose obtained with this method was compared with that obtained at the heavy ion therapy facility HIMAC at the National Institute of Radiological Sciences in Chiba, Japan, using the Japanese code of practice. The agreement found was better than 1%. The combined uncertainty of the determination of absorbed dose to water was estimated to amount to 5%. PMID- 10368013 TI - Reduction of the gamma-ray component from 252Cf fission neutron source- optimization for biological irradiations and comparison with MCNP code. AB - Gamma-rays contribute 33% of the absorbed dose from an unfiltered 252Cf fission neutron source. To reduce this gamma-ray component and to enable radiobiological experiments at as high a dose rate as possible, Monte Carlo calculations for several filter materials (Al, Fe, Pb and concrete) have been made using MCNP neutron and photon transport code version 4a. A lead filter of thickness 4 cm was found to reduce the gamma-ray component to 6.7% of the total dose whilst reducing the neutron dose by only about 10%. Such a filter was installed at the MRC 252Cf neutron irradiation facility and dosimetric measurements were made using a TE-TE chamber and a 7LiF(Mg, Cu, P) TLD. Monte Carlo simulations agree with experimental measurements of neutron and gamma-ray doses within 6%. V79-4 Chinese hamster cells were irradiated with lead-filtered and unfiltered neutrons and also with 60Co gamma-rays at two dose rates. The survival fraction obtained for each radiation was consistent with the reduced gamma-ray dose. The relative biological effectiveness for neutrons alone, corrected for gamma-ray effects, was found to be 9.2 +/- 3.4 from the initial slopes and 3.1 +/- 0.5 at 10% survival, both relative to the acute gamma-rays. PMID- 10368014 TI - An MCNP-based model of a linear accelerator x-ray beam. AB - The Monte Carlo N-Particle radiation transport computer code (MCNP) has been employed on a personal computer to develop a simple model simulating the major components within the beam path of a linear accelerator radiation head, namely the electron target, primary conical collimator, beam flattening filter, wedge filter and the secondary collimators. The model was initially used to calculate the energy spectra and angular distributions of the x-ray beam for the Philips SL 75/5 linear accelerator, in a plane immediately beneath the flattening filter. These data were subsequently used as a 'source' of x-rays at the target position, to assess the emergent beam from the secondary collimators. The depth dose distributions and dose profiles at constant depth for various field sizes have been calculated for a nominal operating potential of 4 MV and found to be within acceptable limits. It is concluded that the technique may be used to calculate the energy spectra of any linear accelerator upon specification of the component dimensions, materials and nominal accelerating potential. It is anticipated that this work will serve as the basis of a quality control tool for linear accelerators and treatment planning systems. PMID- 10368015 TI - An automatic method for the identification and interpretation of clustered microcalcifications in mammograms. AB - We investigated a method for a fully automatic identification and interpretation process for clustered microcalcifications in mammograms. Mammographic films of 100 patients containing microcalcifications with known histology were digitized and preprocessed using standard techniques. Microcalcifications detected by an artificial neural network (ANN) were clustered and some cluster features served as the input of another ANN trained to differentiate between typical and atypical clusters, while others were fed into an ANN trained on typical clusters to evaluate these lesions. The measured sensitivity for the detection of grouped microcalcifications was 0.98. For the task of differentiation between typical and atypical clusters an Az value of 0.87 was computed, while for the diagnosis an Az value of 0.87 with a sensitivity of 0.97 and a specificity of 0.47 was obtained. The results show that a fully automatic computer system was developed for the identification and interpretation of clustered microcalcitications in mammograms with the ability to differentiate most benign lesions from malignant ones in an automatically selected subset of cases. PMID- 10368016 TI - Application of the fundamental parameter method to the in vivo x-ray fluorescence analysis of Pt. AB - The application of the fundamental parameter method (FPM) to the in vivo x-ray fluorescence (XRF) analysis of Pt has been investigated. The FPM is conventionally used to carry out elemental analysis of samples in vitro without the need to use standard samples of accurately known composition for system calibration. The present work has involved the use of the FPM to calculate the concentration of Pt solutions in phantoms, with concentrations ranging from 25 1000 ppm. The phantoms simulate the measurement of Pt-based chemotherapy drugs in head and neck tumours. The radiation sources were a 150 kV tungsten-anode x-ray tube and the isotope 99mTc. The minimum detection limit measured for Pt was in the range 8-30 ppm (depending on radiation source and geometry), using a narrow (5 mm) diameter beam. Dose rates in the phantom were 0.1-5 mGy h(-1). Average differences between nominal and calculated values of Pt concentration were <8% using the phantoms in air to simulate measurement of Pt in superficial body sites. If the phantoms were placed in a water bath, to simulate measurement at greater depths of overlying tissue, higher systematic differences (15-20%) were observed. This effect is probably due to multiple scattering processes in the surrounding medium. PMID- 10368017 TI - Properties of the light emerging from a diffusive medium: angular dependence and flux at the external boundary. AB - By using the diffusion approximation of the radiative transfer equation and the partial-current boundary condition, an analytical expression for the angular dependence of the specific intensity emerging from a diffusive medium has been obtained. The analytical expression for the angular distribution has been validated by comparisons with the results of Monte Carlo simulations. By using the diffusion equation and the extrapolated boundary condition, an heuristic analytical expression for the diffuse time-resolved reflectance has also been obtained by assuming that the photon flux is simply proportional to the fluence rate. For the case of the semi-infinite medium, comparisons with Monte Carlo results are presented and time-resolved reflectance data are fitted with the simple fluence rate formula. The results obtained show that the simple expression correctly describes the time-resolved reflectance giving an error in the retrieved optical parameters smaller than that of other commonly used expressions. PMID- 10368018 TI - Acoustic measurement of compressibility and thermal expansion coefficient of erythrocytes. AB - Mechanical properties of human erythrocytes, namely adiabatic compressibility and thermal expansion coefficient, have been determined using a classical ultrasound velocity and attenuation burst transmission technique. The theoretical model concerns the corpuscular part of the elastic wave propagating in a suspension of viscous particles of small size compared with the wavelength. The thermal wave contribution was taken into account. Normal and stiffened red blood cells were suspended in saline of different NaCl concentration. PMID- 10368019 TI - Measurements of the equivalent whole-body dose during radiation therapy by cytogenetic methods. AB - Estimates of equivalent whole-body dose following partial body exposure can be performed using different biophysical models. Calculations should be compared with biodosimetry data, but measurements are complicated by mitotic selection induced in target cells after localized irradiation. In this paper we measured chromosomal aberrations in peripheral blood lymphocytes during radiotherapy, and estimated the equivalent whole-body dose absorbed, by using the novel technique of interphase chromosome painting. Premature chromosome condensation was induced in stimulated lymphocytes by incubation in calyculin A, and slides were hybridized in situ with whole-chromosome DNA probes specific for human chromosomes 2 and 4. Reciprocal exchanges were used to estimate the equivalent whole-body dose, based on individual pre-treatment in vitro calibration curves. Equivalent whole-body dose increased as a function of the number of fractions, and reached a plateau at high fraction numbers. Chromosomal aberration yields were dependent on field size, tumour position and concurrent chemotherapy. Results suggest that interphase chromosome painting is a simple technique able to give a reliable estimate of the equivalent whole-body dose absorbed during therapeutic partial-body irradiation. PMID- 10368020 TI - Experimental study on the influence of the central electrode in Farmer-type ionization chambers. AB - In the IAEA TRS-381 protocol, k(cel) and p(cel) account for the central electrode perturbation during the air kerma chamber calibration and the in-phantom measurements. The values of these correction factors are based mainly on Monte Carlo simulations. In the present work experimental data on k(cel) and p(cel) for the NE-2571 chamber is provided, relative to a graphite electrode. In addition, the relative influence of the 3 mm diameter A-150 central electrode of the NE 2581 chamber is studied. The experimentally determined value of k(cel) for a 1 mm aluminium electrode is 1.008 +/- 0.2%, and of p(cel) in photon and electron beams 0.993 +/- 0.2% and 0.997 +/- 0.2% respectively. The experimental data and the Monte Carlo simulations agree to within 0.2%. No significant influence of the A 150 central electrode diameter on the absorbed dose determination is shown. PMID- 10368021 TI - A study on virtual source position for electron beams from a Mevatron MD linear accelerator. AB - The virtual source position (VSP) for electron beams of energies 5, 7, 9 10, 12 and 14 MeV and for the applicators (cones) available in the department have been measured for a Mevatron MD class linear accelerator. Different methods of obtaining the virtual source position for electron beams have been investigated in the present study. The results obtained have been compared with those of other workers. It is observed that the VSP is very much machine dependent and needs to be measured for each linear accelerator. The effect of shielding on virtual source position for the type of applicators available in the department has also been investigated. PMID- 10368023 TI - Proportionality between Wiener spectra of quantum mottle and the squares of modulation transfer functions. AB - Rossmann proposed that the Wiener spectra of the quantum mottle of radiographs made using screen-film systems were proportional to the squares of the modulation transfer functions (MTFs) of the screen-film systems. On the other hand, Lubberts theoretically pointed out that the shape of the Wiener spectrum of the quantum mottle depended on the sum of the squares of the MTFs for different depths in the screen phosphor layer, rather than the square of the sum of the MTFs for the different depths, i.e. the square of the MTF of the screen-film systems. The purpose of this study is to experimentally investigate the proportionality between the Wiener spectra of the quantum mottle and the squares of the MTFs of screen-film systems using two screen-film systems having different screen thicknesses. For this purpose, we determined correction factors for the square of the MTF of the screen-film system in the Wiener spectrum of the quantum mottle at each spatial frequency when the Wiener spectral values of the screen mottle were separated into those of the quantum mottle and structure mottle. The correction factor is the ratio of the normalized Wiener spectrum of the quantum mottle to the square of the MTF of the screen-film system. As a result, for a thin screen, the correction factors were unity for all spatial frequencies; on the contrary, for a thick screen, the factor increased with spatial frequency. By calculating the theoretical correction factors using the models for the MTF and Wiener spectrum of the quantum mottle of Nishikawa and Yaffe based on Lubberts' theory, we verified that our experimental results agreed with Lubberts' theory. Furthermore, by obtaining the screen thickness dependence of the theoretical correction factors for the two screens, we showed that, for screens thinner than 0.02 mm, Rossmann's theory can be applied to the relationship between the Wiener spectrum of the quantum mottle and the MTF of the screen-film system, whereas for screens thicker than 0.02 mm, Lubberts' theory should be applied. PMID- 10368022 TI - Photon-counting radiography with the gas microstrip detector. AB - We have built a proof of-concept photon-counting x-ray imaging system using a Xe:CH4 gas microstrip detector (GMD) as the image receptor, and have used this system to demonstrate several advantages of photon counting over energy integration. Our experiments spanned x-ray spectra from 10 to 50 kVp and Xe:CH4 pressures from 1 to 4 atm. When photon counting is done, the energy deposited in the detector by each incident photon can be measured on adjacent anode strips and a centroid calculation can be used to provide spatial resolution significantly better than the anode strip pitch. We measured > 11 lp mm(-1) at 13 kVp with our 200 microm pitch detector, and 8.2 lp mm(-1) at 50 kVp. The energy resolution of our GMD is 5.2% at 59.6 keV, and the space-charge limited counting rate is >2 x 10(6) mm(-2) s(-1) at 3 atm for a 30 kVp beam. PMID- 10368024 TI - A simple model for estimating the particle size dependence of absolute efficiency of fluorescent screens. AB - The absolute efficiency of a phosphor screen is the ratio of the light energy per unit area at the screen surface to the incident x-ray energy fluence. Particle size is a critical factor in determining the absolute efficiency, but in most models its influence is not accounted for. To allow derivation of the particle size dependence, a model is proposed that describes the optical properties of the screen by means of a single parameter, the light extinction factor, xi, and assumes that the intrinsic efficiency (light energy/energy imparted to the phosphor material) is independent of particle size. The value of xi depends on the type of screen (phosphor, reflective backing, coating and binder) and has to be determined from measurements on at least two screens with known particle size and thickness. The absolute efficiency can then be calculated for an extended range of particle sizes and/or screen thicknesses. To test the model, experimental data from the literature were used to derive values of xi for screens of La2O2S:Tb, LaOBr:Tm and ZnCdS:Ag. The extinction factor was found to vary between -6 and +20%. The non-physical negative value for xi, found from one set of experiments on La2O2S:Tb screens, may be explained as resulting from a lack of accurate knowledge of the actual tube potential, influencing calculated values of the energy imparted to the screen. The results are promising but further well-controlled experiments (including improved dosimetric calculations to account forescape of K-radiation from the screen) are needed to confirm the model. PMID- 10368026 TI - Subsets and overrelaxation in iterative image reconstruction. AB - A number of iterative image reconstruction algorithms were integrated into one formula characterizing each algorithm by only two parameters: overrelaxation and number of subsets. From the formula it follows that the ordered-subsets iteration (OS-EM) is equivalent to iteration with overrelaxation, where the OS level corresponds to the overrelaxation parameter. Algorithms represented by the formula were studied with respect to speed of convergence and image characteristics. In particular, OS-EM was compared with a single-projection iteration procedure using an optimized sequence of overrelaxation parameters (HOSP) which combines rapid convergence with reduced storage requirements. As a result, OS-EM with a constant number of subsets either needed more iteration steps than HOSP or provoked additional noise, depending on the number of subsets used during iteration. OS-EM can be improved by using decreasing OS levels, imitating the decreasing overrelaxation parameters used for HOSP. The resulting OS-EM may be slightly more rapid than HOSP, due to the increasing number of projections used simultaneously. PMID- 10368025 TI - Detective quantum efficiency of an amorphous selenium detector to megavoltage radiation. AB - The spatial frequency dependent detective quantum efficiency (DQE(f)) of a high resolution selenium-based imaging system has been measured at megavoltage energies. These results have been compared with theoretical calculations. The imaging system was a video tube with a 5 microm amorphous selenium (a-Se) target which was irradiated by 1.25 MeV gamma-rays. The modulation transfer function (MTF) decreased rapidly with spatial frequency (determined by spread of electrons in the build-up material) while the noise power spectrum was constant as a function of spatial frequency. The DQE obtained from these MTF and noise power measurements was compared with a Monte Carlo model of the pulse height spectrum of the detector. The DQE(0) model accounted for the interaction of x rays with the detector as well as the energy-dependent gain (charge generated/energy deposition). Good agreement between the calculated and measured DQE(0) was found. The model was also used to estimate the DQE(f) of a metal plate + a-Se detector which was compared with a metal plate + phosphor system of the same mass thickness. The DQE(f) s of both detectors are very similar, indicating that the choice of which detector is better will be based upon criteria other than DQE(f), such as read-out approach, ease of manufacture or sensitivity. PMID- 10368027 TI - Non-invasive measurement of chemotherapy drug concentrations in tissue: preliminary demonstrations of in vivo measurements. AB - Measurements of the tissue concentrations of two chemotherapy agents have been made in vivo on an animal tumour model. The method used is based on elastic scattering spectroscopy (ESS) and utilizes a fibre-optic probe spectroscopic system. A broadband light source is used to acquire data over a broad range of wavelengths and, therefore, to facilitate the separation of absorptions from various chromophores. The results of the work include measurements of the time course of the drug concentrations as well as a comparison of the optical measurements with high performance liquid chromatography (HPLC) analysis of the drug concentrations at the time of sacrifice. It is found that the optical measurements correlate linearly with HPLC measurements, but give lower absolute values. PMID- 10368028 TI - A novel frequency domain fluorescence technique for determination of triplet decay times. AB - Frequency domain fluorescence measurement using two diode lasers with amplitude modulation in the kHz range yields a signal component at the sum frequency. This intermodulation phenomenon was observed in an aqueous solution of haematoporphyrin (HP) and could be related to triplet state population kinetics. This indirect measurement technique may allow triplet decay time measurement during photodynamic therapy (PDT) enabling monitoring of the type II phototoxic damage rate. PMID- 10368029 TI - Optimization of axial RF field distribution in low-frequency EPR loop-gap resonators. AB - A novel coupling method that optimizes the axial RF distribution of low-frequency EPR loop-gap resonators is presented. It consists of a resonant coupling loop positioned at the centre of a two-section loop-gap resonator. This arrangement ensures a symmetrical distribution of the radio frequency field along the axis of the resonator. The design of a central coupling system suitable for EPR resonators operating at about 220 MHz is described. Experimental results show that with the central coupling system the RF field is symmetrical and has a very good axial homogeneity (100% of the resonator length). PMID- 10368030 TI - Comments on 'Investigation of the chamber correction factor (k(ch)) for the UK secondary standard ionization chamber (NE2561/NE2611) using medium energy x rays'. PMID- 10368031 TI - Endogenous and exogenous factors on sleep-wake cycle regulation. AB - A number of theories have proposed the involvement of different brain structures and neurotransmitters in order to explain the regulation of the sleep wake cycle. However, there is no clear consensus as to the mechanisms through which the brain structures and their various neurotransmitters interact to produce theses phases. Perhaps the problem is related to the fact sleep is a very fragile state, easily modified or influenced by a variety of substances or experimental manipulations. In this paper, we describe the evidence of two different groups of factors that induce important changes on the sleep wake cycle. The endogenous factors: neurotransmitters; hormone; peptides; and some substances of lipidic nature and exogenous factors: stress, food intake, learning, sleep deprivation, sensorial stimulation, exercise and temperature on the regulation the sleep-wake cycle. Likewise, we propose a hypothesis which attempts to reconcile the fact that endogenous and exogenous factors have similar effects. PMID- 10368032 TI - The effects of cannabinoids on the brain. AB - Cannabinoids have a long history of consumption for recreational and medical reasons. The primary active constituent of the hemp plant Cannabis sativa is delta9-tetrahydrocannabinol (delta9-THC). In humans, psychoactive cannabinoids produce euphoria, enhancement of sensory perception, tachycardia, antinociception, difficulties in concentration and impairment of memory. The cognitive deficiencies seem to persist after withdrawal. The toxicity of marijuana has been underestimated for a long time, since recent findings revealed delta9-THC-induced cell death with shrinkage of neurons and DNA fragmentation in the hippocampus. The acute effects of cannabinoids as well as the development of tolerance are mediated by G protein-coupled cannabinoid receptors. The CB1 receptor and its splice variant CB1A, are found predominantly in the brain with highest densities in the hippocampus, cerebellum and striatum. The CB2 receptor is found predominantly in the spleen and in haemopoietic cells and has only 44% overall nucleotide sequence identity with the CB1 receptor. The existence of this receptor provided the molecular basis for the immunosuppressive actions of marijuana. The CB1 receptor mediates inhibition of adenylate cyclase, inhibition of N- and P/Q-type calcium channels, stimulation of potassium channels, and activation of mitogen-activated protein kinase. The CB2 receptor mediates inhibition of adenylate cyclase and activation of mitogen-activated protein kinase. The discovery of endogenous cannabinoid receptor ligands, anandamide (N arachidonylethanolamine) and 2-arachidonylglycerol made the notion of a central cannabinoid neuromodulatory system plausible. Anandamide is released from neurons upon depolarization through a mechanism that requires calcium-dependent cleavage from a phospholipid precursor in neuronal membranes. The release of anandamide is followed by rapid uptake into the plasma and hydrolysis by fatty-acid amidohydrolase. The psychoactive cannabinoids increase the activity of dopaminergic neurons in the ventral tegmental area-mesolimbic pathway. Since these dopaminergic circuits are known to play a pivotal role in mediating the reinforcing (rewarding) effects of the most drugs of abuse, the enhanced dopaminergic drive elicited by the cannabinoids is thought to underlie the reinforcing and abuse properties of marijuana. Thus, cannabinoids share a final common neuronal action with other major drugs of abuse such as morphine, ethanol and nicotine in producing facilitation of the mesolimbic dopamine system. PMID- 10368033 TI - CCK-B receptor: chemistry, molecular biology, biochemistry and pharmacology. AB - Cholecystokinin (CCK) is a peptide originally discovered in the gastrointestinal tract but also found in high density in the mammalian brain. The C-terminal sulphated octapeptide fragment of cholecystokinin (CCK8) constitutes one of the major neuropeptides in the brain; CCK8 has been shown to be involved in numerous physiological functions such as feeding behavior, central respiratory control and cardiovascular tonus, vigilance states, memory processes, nociception, emotional and motivational responses. CCK8 interacts with nanomolar affinities with two different receptors designated CCK-A and CCK-B. The functional role of CCK and its binding sites in the brain and periphery has been investigated thanks to the development of potent and selective CCK receptor antagonists and agonists. In this review, the strategies followed to design these probes, and their use to study the anatomy of CCK pathways, the neurochemical and pharmacological properties of this peptide and the clinical perspectives offered by manipulation of the CCK system will be reported. The physiological and pathological implication of CCK-B receptor will be confirmed in CCK-B receptor deficient mice obtained by gene targeting (Nagata el al., 1996. Proc. Natl. Acad. Sci. USA 93, 11825-11830). Moreover, CCK receptor gene structure, deletion and mutagenesis experiments, and signal transduction mechanisms will be discussed. PMID- 10368034 TI - Brain shrinkage in alcoholics: a decade on and what have we learned? AB - Brain atrophy in alcoholics has been identified using both radiological and pathological techniques. However the magnitude and topography of the atrophy, and the factors which contribute to it, are unclear. This review compares the results of imaging and pathological studies in alcoholics examining variables which may contribute to any discrepancies. We conclude that significant brain damage does occur as a result of alcohol abuse per se, that the damage is regionally specific with the frontal lobes being particularly affected, and that both grey matter and white matter components are damaged. PMID- 10368035 TI - Relative biological effectiveness of proton beams in clinical therapy. AB - PURPOSE: In clinical proton beam radiation therapy, an RBE of 1.1 relative to megavoltage X-rays is currently being employed at most treatment centers. This RBE pertains to radiation in the spread out Bragg-peak (SOBP) for all tissue systems, all dose levels per fraction and all proton beam energies. As the number of centers and treatment sites for which proton beam therapy continues to increase and additional experimental data is accrued, a re-assessment of the justification for a generic RBE is warranted. In this paper we address: (1) the constancy of the RBE along the central axis from the plateau entrance to the distal SOBP (upstream of the distal edge); (2) RBE as a function of dose (or cell survival level); and (3) the target cell or tissue (alpha/beta) dependency of the RBE. This analysis pertains to modulated proton beams of initial energies of approximately 70-200 MeV and SOBPs of approximately 2-10 cm, respectively. RESULTS AND CONCLUSIONS: With exceptions, the available experimental data indicate that the RBE of SOBP protons increases with decreasing dose or dose per fraction and increasing depth in the SOBP, with the magnitude of both effects likely being dependent on the alpha/beta ratios of the target cells or tissues. The use of a generic RBE of 1. for all tissues, especially those exhibiting low alpha/beta values such as CNS, may be too low, especially at dose levels of < or = 2 Gy/fraction. Systematic determination of the RBE values dependent upon the three interdependent variables identified in this manuscript (beam depth, dose size and target tissue) will provide an enhanced data base for detailed treatment planning and institutional trial comparisons, thereby maximizing the therapeutic benefit of proton beams. PMID- 10368036 TI - Stereotactically guided conformal radiotherapy: a new tool for the treatment of intra-cranial benign tumors. PMID- 10368038 TI - Differences in target outline delineation from CT scans of brain tumours using different methods and different observers. AB - PURPOSE: To assess errors resulting from manual transfer of contour information for three-dimensional (3-D) target reconstruction, and to determine variations in target volume delineation of brain tumours by different radiation oncologists. MATERIALS AND METHODS: Images of 18 patients with intracranial astrocytomas were used for retrospective treatment planning by five radiation oncologists. In this study, the target outline was delineated on sequential CT slices by an experienced radiation oncologist. Thereafter, the target outline was manually reconstructed by five radiation oncologists onto an A-P or lateral scout film. The same target outline was also reconstructed as a projection using the Beam's eye view capability on a CT simulator unit. The two target outlines were compared by encompassing each shape with the smallest rectangle. The manually reconstructed radiation field was termed 'Field manually established on X-ray film (F-X)', and the automatically-established field was termed 'Field established by CT simulator (F-CT)'. In a second part of this study, four radiation oncologists defined contours from contrast enhanced CT images of nine patients with intracranial astrocytomas. The CT images of these nine cases included five pre-operative cases and four post-operative cases. Both gross tumour volume (GTV) and clinical target volume (CTV) were outlined on sequential CT slices. The target outlines for the four radiation oncologists were compared by identifying the smallest rectangular field surrounding the projection of these contours. The field established by each radiation oncologist was termed 'Field of target volume (F-TV)', and the overlapping portion of the four F-TVs for each case was termed 'Overlapped field of the target volume (Fo-TV)'. RESULTS: The average distance between the isocentres of F-X and F-CT was 0.6 +/- 0.4 cm (mean +/- SD). The average ratio of the area of F-X divided by the area of F-CT was 1.04 +/- 0.12. The area of F-X was wider than the area of F-CT for four of the five oncologists. The ratio of the area of F-TV divided by the area of Fo-TV was calculated. The average ratio was relatively greater for CTV (2.07 in pre operative cases and 2.11 in post-operative cases) than for GTV (1.12 in pre operative cases and 1.41 in post-operative cases). Among radiation oncologists, variations in the delineation of GTV were smaller than those of CTV. CONCLUSIONS: When using an X-ray simulator in treatment planning, errors resulting from the manual transfer of CT contour information to planar radiographs must be considered. When computer techniques are used to project contours onto radiographs errors resulting from individual variations when performing the contouring must be considered. PMID- 10368037 TI - Stereotactically guided conformal radiotherapy for meningiomas. AB - PURPOSE: Stereotactically guided conformal radiotherapy, (SCRT) is a high precision technique of conformal radiotherapy (RT) which reduces the volume of normal tissue irradiated compared to conventional RT and may lead to a reduction in long-term toxicity We describe the technique and the preliminary results in patients with inoperable, residual or recurrent meningiomas. MATERIAL AND METHODS: From July 1993 to November 1997, 24 patients (median age: 56 years, range: 28-72) with base of skull (n = 21). falx or upper skull (n = 3) meningiomas were treated with SCRT. The technique employed immobilization in a Gill-Thomas-Cosman (GTC) frame and CT localization with a Brown-Roberts-Wells (BRW) fiducial system for stereotactic space definition. The planning target volume (PTV) was defined as gross tumour volume (GTV) and a 0.5-1 cm margin. Treatment was delivered with three (12 patients) or four non-coplanar conformal fixed fields (12 patients) Conformal blocking was achieved either with lead alloy blocks (n = 11) or with a multi-leaf collimator (MLC) (n = 13). Patients were treated on a 6 MV linear accelerator to doses of 50-55 Gy, in 30-33 daily fractions. The treatments were carried out as part of a routine work of a busy radiotherapy department. RESULTS: Median GTV for 24 meningiomas was 21.7 cm3 (range: 4.4-183 cm3). SCRT was well tolerated with minimal toxicity Three months after the end of radiotherapy, seven of 15 patients with neurological deficit had an improvement and eight remained unchanged. Two patients experienced early side effects (one VII nerve palsy, one Addisonian state). At a median follow-up of 13 months (range: 3-43) the 1 year progression free survival and overall survival are 100%. which is within the range expected for conventional fractionated radiotherapy for meningiomas. CONCLUSIONS: SCRT is a feasible technique of high precision conformal RT for patients with meningiomas. Potential advantages in tumour control, survival and toxicity over conventional RT, require evaluation in long-term prospective studies. PMID- 10368039 TI - Normobaric oxygen treatment during radiotherapy for carcinoma of the uterine cervix. Results from a prospective controlled randomized trial. AB - BACKGROUND AND PURPOSE: Hypoxia, a frequent characteristic of cervical cancer, is associated with reduced sensitivity to irradiation and thus may be a source of radiotherapy failure. This study was planned to test the hypothesis, that inhalation of oxygen during radiotherapy may increase the radiation effect on the tumor and improve loco-regional control and overall survival. MATERIAL AND METHODS: From 1963 to 1965, a consecutive series of 208 patients with cervical cancer stage II/III who were to be treated by external irradiation plus radium inserts, were included in this study. They were randomly assigned to either receive oxygen inhalations during the radiotherapy sessions or just breathing air. Due to technical reasons the oxygen group was divided. For the first 10 months, they did receive oxygen during the radium inserts only, the last 13 months during all radiotherapy sessions. RESULTS: After median 33 years follow up, there are no differences in overall survival, cancer-specific survival or loco-regional control. Subgroup analysis shows significantly improved loco regional control in the stage IIB patients, with squamous cell carcinoma who received oxygen during all radiotherapy sessions. This improvement was especially pronounced among the patients who also received blood transfusions. CONCLUSIONS: There was no influence of normobaric oxygen treatment on the overall outcome to radiotherapy in patients with stage II cervical cancer, but subgroup analyses support the hypothesis that there is tumor areas of hypoxia-based radioresistance that may be counteracted by oxygen administration. PMID- 10368040 TI - Improvement in human tumour oxygenation with carbogen of varying carbon dioxide concentrations. AB - BACKGROUND AND PURPOSE: Carbogen (95%O2, 5%CO2) is being used in clinical trials as a hypoxic radiosensitiser. Tolerance to carbogen can be a problem, this study compares tumour oxygenation during inhalation of hyperoxic gas containing either 2% or 5% CO2. MATERIALS AND METHODS: Tumour pO2 was measured in 16 patients using the Eppendorf pO2 histograph. RESULTS: After breathing gas containing either 5% or 2% CO2 an increase in median pO2 was measured in every tumour, the frequency of low pO2 values ( < or = 10 mmHg) fell from 47% to 29% in the 5% group and from 55% to 17% in the 2% group. CONCLUSIONS: This study confirms that breathing 2% CO2 and 98% O2 is well tolerated and effective in increasing tumour oxygenation. PMID- 10368041 TI - Vascular architecture and microenvironmental parameters in human squamous cell carcinoma xenografts: effects of carbogen and nicotinamide. AB - BACKGROUND AND PURPOSE: A better understanding of the vascular architecture and the microenvironmental parameters (VAMP) will allow the identification of tumours that can be more effectively treated by intensified fractionated radiotherapy or modifiers of blood flow and oxygenation or combinations of these approaches. MATERIALS AND METHODS: Proliferation (BrdUrd), vascular architecture (endothelial marker), perfusion (Hoechst 33342) and oxygenation (NITP) were studied in two human laryngeal squamous cell carcinoma tumour lines grown as xenografts in nude mice. The effects of carbogen and nicotinamide on these parameters were evaluated. RESULTS: Carbogen treatment resulted in a decrease of the number of perfused blood vessels from 66% to 55% in one of the two tumour lines. In this tumour line nicotinamide prevented this reduction of tumour blood flow by carbogen. In both tumour lines the labelling index (LI) decreased after treatment with carbogen for 1 h, from 11-13% to 5-7%. Both tumour lines showed a drastic reduction of hypoxia by carbogen alone or by carbogen plus nicotinamide. CONCLUSIONS: In both laryngeal squamous cell carcinoma xenograft tumour lines carbogen was very effective in reducing diffusion limited hypoxia. Only in one of the two tested tumour lines carbogen also caused a reduction of tumour blood perfusion, which could be compensated for by nicotinamide. In addition, carbogen reduced tumour cell proliferation. The fact that differences in response to nicotinamide and carbogen were observed and that they can be studied in vivo provides a basis for further development of a 'predictive profile' which will guide the clinician to select the optimal treatment for individual patients or groups of patients. PMID- 10368042 TI - Erythropoietin for patients undergoing radiotherapy: a pilot study. AB - BACKGROUND AND PURPOSE: To evaluate the feasibility and efficacy of using recombinant human erythropoietin (rhEPO) to correct decreased hemoglobin levels in patients undergoing radiotherapy and to get an estimate of its influence on the efficacy of radiotherapy. MATERIALS AND METHODS: Fifty patients with cancer of the head and neck and the pelvis were randomized before radiotherapy to different rhEPO treatments (none, 3 x 150 U/kg per week i.v., 3 x 300 U/kg per week i.v. and 3 x 150 U/kg per week s.c.). Hematological parameters were evaluated weekly and the locoregional tumor control rates were determined in 38 patients with head and neck cancer. RESULTS: rhEPO-treated patients showed a significant increase in their hemoglobin values (0.7 g/100 ml per week). The rhEPO response was comparable for patients with cancer of the head and neck and the pelvis. A delayed recovery was seen when iron deficiency or impaired iron mobilization was present. No serious toxicity was observed. Locoregional tumor control was improved, although not statistically significantly, in those head and neck cancer patients who experienced a rapid rise of hemoglobin. CONCLUSIONS: Low hemoglobin levels can be safely and quickly corrected with rhEPO. This may improve the effectiveness of radiotherapy. PMID- 10368043 TI - Second non-germ cell malignancies in patients treated for stage I-II testicular seminoma. AB - PURPOSE: To measure the incidence of second non-germ cell malignancies (SNGCM) in patients treated for a stage I-II testicular seminoma. MATERIALS AND METHODS: From 1970 to 1992, 131 evaluable patients received in the Institut Claudius Regaud a post-orchiectomy treatment for a stage I-II testicular seminoma. The therapeutic modalities, including salvage treatment for six recurrences, were as follows: infradiaphragmatic radiotherapy (IDRT) (n = 55); infra- and supradiaphragmatic radiotherapy (IDRT + SDRT) (n = 64); IDRT + SDRT with chemotherapy (n = 12). The mean follow-up was 11 years. The cumulative incidence of SNGCM was compared to the overall cancer incidence in the general male population on the basis of the Tarn Cancer Registry; the relative risk was expressed as a standardized incidence ratio (SIR). RESULTS: Overall, the cumulative incidence of SNGCM was 10.7% (14/131 cases). The SIR was equal to 2.81 (95% confidence interval (CI) 1.54-4.72; P < 0.001) and increased with follow-up duration. The SIR was significantly increased in 64 patients treated with IDRT + SDRT (SIR = 3.08; 95% CI 1.47-5.66; P = 0.002) but not in 55 patients treated with IDRT alone (SIR = 0.62; 95% CI 0.01-3.43; P = 0.8). The 12 patients who received chemotherapy had an SIR of 26.2 (95% CI 5.48-77.69; P < 0.001), while the SIR was 2.26 in the 119 patients who did not receive any chemotherapy (95% CI 1.13-4.04; P = 0.01 ). Of four hematologic malignancies, three appeared in the 12 patients who received chemotherapy. CONCLUSIONS: An increased risk of SNGCM after SDRT + IDRT has been demonstrated. After IDRT alone, the risk of second cancer is not incremented after a median follow-up of 6 years, but further observation of the patients is necessary to achieve final conclusions. Our results suggest that the risk of second cancer and especially of hematologic malignancy is increased by the association of chemotherapy and radiation. PMID- 10368044 TI - Impact of localized radiotherapy on blood immune cells counts and function in humans. AB - Immune cells subsets were prospectively analyzed after localized radiotherapy (LRT). LRT reduced the levels of all lymphocyte subsets, with B-cells and naive T cells being most sensitive. Lymphocyte function was suppressed, but still within the normal range. Rapid recovery of cytotoxic T-cells/natural killer cells after LRT and the functional suppression within normal levels explains the low incidence of infections after LRT. PMID- 10368045 TI - Upregulation and spatial shift in the localization of the mannose 6 phosphate/insulin-like growth factor II receptor during radiation enteropathy development in the rat. AB - BACKGROUND AND PURPOSE: Transforming growth factor beta1 (TGF-beta1) appears to play an important role in the pathogenesis of chronic radiation-induced fibrosis in the intestine and several other organs. TGF-beta1 is secreted as a non biologically active complex and its function depends on activation. In vitro data suggest that the mannose 6-phosphate/insulin-like growth factor-beta (M6P/IGF-II) receptor is involved in the mechanism of TGF-beta1 activation. Thus, we used a rat model of radiation enteropathy to examine the potential role of the M6P/IGF II receptor in the in vivo regulation of TGF-beta1 activity and localization. MATERIALS AND METHODS: A scrotal hernia containing a loop of small intestine was created in male rats. The intestine in the scrotum was exposed to 0, 12, or 21 Gy single dose X-radiation. Groups of rats were euthanized 1 day and 2, 6 and 26 weeks after irradiation. Histopathologic injury was assessed with a radiation injury score (RIS). Computerized image analysis was used to identify M6P/IGF-II receptor-positive cells and to quantify extracellular matrix-associated TGF-beta1 immunoreactivity. Changes in urokinase plasminogen activator (uPA), tissue-like plasminogen activator (tPA) and plasminogen activator inhibitor-1 (PAI-1) immunoreactivity were also assessed. RESULTS: In normal (sham-irradiated) intestine, M6P/IGF-II immunoreactivity was confined to relatively weak, but specific epithelial staining. Irradiated intestine exhibited a highly significant time- and dose-dependent increase in the number of M6P/IGF-II receptor-positive cells (P < 0.001). There was a striking spatial shift of M6P/IGF-II receptor immunoreactivity from epithelium during the early post-radiation phase to stromal cells, most notably fibroblasts during the later stages of injury. Irradiated intestine exhibited distinct co-localization of M6P/ IGF-II receptor-positive cells and extracellular matrix-associated TGF-beta1 in areas of histopathologic injury. There were highly significant associations between the number of M6P/IGF II receptor-positive stromal cells and TGF-beta1 immunoreactivity (P < 0.001), radiation-induced fibrosis (P < 0.001) and RIS (P < 0.001). Endothelial tPA immunoreactivity decreased significantly after irradiation (P < 0.001), whereas uPA and PAI-1 immunoreactivity levels appeared to be unchanged. CONCLUSIONS: M6P/IGF-II receptor upregulation may be a key factor in the in vivo control of TGF-beta1 activity and responsible for the tissue specificity of TGF-beta1 action after irradiation. PMID- 10368046 TI - Radiosensitization by intratumoral administration of cisplatin in a sustained release drug delivery system. AB - PURPOSE: Effects of combining local irradiation and intratumoral (i.t.) administration of cisplatin (CDDP) in a sustained-release drug delivery system (epi gel) were studied in a murine SCCVII squamous cell carcinoma model in mice. MATERIALS AND METHODS: The epinephrine injectable gel was used as a drug delivery system. Intratumoral pharmacokinetics of CDDP was studied by using 195mPt-CDDP. The tumor volume quadrupling time (TVQT) and tumor growth delay (TGD) time were used to evaluate the antitumor efficacy of treatment regimens. RESULTS: The concentration and residence of 195mPt-CDDP was significantly higher in tumors treated with 195mpt-CDDP/epi gel than in tumors treated with 195mPt CDDP gel or 195mPt-CDDP suspension. Intratumoral administration of CDDP/epi gel (4 mg/kg) produced an average TGD time of 15.5 +/- 2.8 days, which was 5.2 - 7.4 times longer than CDDP suspension i.t. or i.p. When combined with a single dose of radiation (10 Gy), i.t. administration of CDDP/epi gel was 2.0 - 3.6-fold as effective as administered i.t. in suspension (39.2 +/- 4.1 vs. 19.8 +/- 3.9 days of TGD, P < 0.05) or i.p. in solution (39.2 +/- 4.1 vs. 11.0 +/- 1.6 days, P < 0.001) in inhibiting tumor growth and produced 20-60% complete remission of tumors. When combined with fractionated irradiation, pre-irradiation CDDP administration was more effective than post-radiation administration (26.7 vs. 12.1 days of TGD, P < 0.05). Mice treated with CDDP/epi gel i.t. alone or in combination with irradiation, had little systemic toxicity. CONCLUSIONS: Intratumoral administration of CDDP using the sustained-release drug delivery system is an efficient and safe method to maximize the drug concentration in tumor, minimize the systemic toxicity and enhance antitumor efficacy of irradiation. PMID- 10368047 TI - Quantification and predictors of prostate position variability in 50 patients evaluated with multiple CT scans during conformal radiotherapy. AB - PURPOSE: To determine the extent and predictors for prostatic motion in a large number of patients evaluated with multiple CT scans during radiotherapy, and evaluate the implications of these data on the design of appropriate treatment margins for patients receiving high-dose three-dimensional conformal radiotherapy. MATERIALS AND METHODS: Fifty patients underwent four serial computerized tomography (CT) scans, consisting of an initial planning scan and subsequent scans at the beginning, middle, and end of the treatment course. Each scan was performed with the patient in the prone treatment position within an immobilization device used during therapy. Contours of the prostate and seminal vesicles were drawn on the axial CT slices of each scan, and the scans were matched by alignment of the pelvic bones with a chamfer matching algorithm. Using the contour information, distributions of the displacement of the organ center of mass and organ border from the planning position were determined separately for the prostate and seminal vesicles in each of the three principle directions: anterior-posterior (AP), superior-inferior (SI) and left-right (LR). Each distribution was fitted to a normal (Gaussian) distribution to determine confidence limits in the center of mass and border displacements and thereby evaluate for the optimal margins needed to contain target motion. RESULTS: The most common directions of displacement of the prostate center of mass (COM) were in the AP and SI directions and were significantly larger than any LR movement. The mean prostate COM displacement (+/- 1 standard deviation, SD) for the entire population was -1.2 +/- 2.9 mm, -0.5 +/- 3.3 mm and -0.6 +/- 0.8 mm in the, AP and SI and LR directions respectively (negative values indicate posterior, inferior or left displacement). The mean (+/- 1 SD) seminal vesicle COM displacement for the entire population was - 1.4 +/- 4.9 mm, 1.3 +/- 5.5 mm and 0.8 +/- 3.1 mm in the AP and SI and LR directions, respectively. The data indicate a tendency for the population towards posterior displacements of the prostate from the planning position and both posterior and superior displacements of the seminal vesicles. AP movement of both the prostate and seminal vesicles were correlated with changes in rectal volume (P = 0.0014 and < 0.0001, respectively) more than with changes in bladder volume (P = 0.030 for seminal vesicles and 0.19 for prostate). A logistic regression analysis identified the combination of rectal volume > 60 cm3 and bladder volumes > 40 cm3 as the only predictor of large ( > 3 mm) systematic deviations for the prostate and seminal vesicles (P = 0.05) defined for each patient as the difference between organ position in the planning scan and mean position as calculated from the three subsequent scans. CONCLUSIONS: Prostatic displacement during a course of radiotherapy is more pronounced among patients with initial planning scans with large rectal and bladder volumes. Such patients may require more generous margins around the CTV to assure its enclosure within the prescription dose region. Identification and correction of patients with large systematic errors will minimize the extent of the margin required and decrease the volume of normal tissue exposed to higher radiation doses. PMID- 10368048 TI - A horizontal CT system dedicated to heavy-ion beam treatment. AB - A horizontal helical CT system to be used for 3-D treatment planning and for positioning verification of patients in seated position was installed in the treatment room with a fixed horizontal heavy-ion beam line. The system achieved the expected mechanical consistency and reliability. PMID- 10368049 TI - Patient position reproducibility in fractionated stereotactic radiotherapy: an update after changing dental impression material. AB - The reproducibility of patient positioning within the Gill-Thomas-Cosman relocatable stereotactic frame was re-evaluated following the substitution of a new, softer, dental impression material for the original hard acrylic compound. The average total displacement for a series of 10 patients was 1.1 mm (+/- 0.6 mm). Rotational discrepancies were small. This technique cannot deliver accurate repositioning in the absence of patient co-operation. PMID- 10368050 TI - Aerosol and splatter production by focused spray and standard ultrasonic inserts. AB - BACKGROUND: Control of contamination in the dental office has sometimes deterred practitioners from using ultrasonic scalers. Recent studies point to the aerosol and splatter produced during ultrasonic scaling as a vehicle for the possible transmission of bloodborne pathogens. A recently introduced ultrasonic insert that focuses the spray produced during scaling may reduce this aerosol contamination. An aerosol reduction device (ARD) that is attached to the ultrasonic handpiece has been shown to reduce the contamination cloud by placing suction in close proximity to the ultrasonic tip. The purpose of this study was to compare the contamination produced by a standard insert (S) and the new focused spray (F) insert with and without the use of the aerosol reduction device (ARD). METHODS: The testing was conducted in vitro within a plastic enclosure using a dye in the coolant spray. After mock scaling of a dentoform model, the number of contaminated squares on the enclosure was counted and recorded. RESULTS: Analysis of the data indicated no significant difference (P >0.05, Mann Whitney U test) between the S or F inserts in the amount of contamination produced. When the aerosol reduction device was used, there was a significant reduction (P <0.05, Mann-Whitney U test) in the amount of contamination for both inserts with a greater reduction for the standard insert. CONCLUSIONS: The traditional style of ultrasonic insert (S) and the newer focused coolant water insert (F) produce an equal amount of aerosol contamination. The amount of aerosol contamination produced by both inserts is copious. The ARD significantly reduced contamination with both styles of inserts. These findings support the use of a large bore high-volume evacuator whenever an ultrasonic scaler is used. PMID- 10368051 TI - Relationship between herpesviruses and adult periodontitis and periodontopathic bacteria. AB - BACKGROUND: Various mammalian viruses and specific bacteria seem to play important roles in the pathogenesis of human periodontitis. This study examined the relationship between subgingival herpesviruses and periodontal disease and potential periodontopathic bacteria in 140 adults exhibiting either periodontitis or gingivitis. METHODS: A nested-polymerase chain reaction (PCR) method determined the presence of Epstein-Barr virus type 1 and type 2 (EBV-1, EBV-2), human cytomegalovirus (HCMV), and herpes simplex virus (HSV) and a 16S rRNA PCR detection method identified Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Bacteroides forsythus, Prevotella intermedia, Prevotella nigrescens, and Treponema denticola. RESULTS: Using a logistic analysis, EBV-1 showed significant positive association with P. gingivalis (odds ratio [OR] 3.37), and with coinfections of P. gingivalis and P. intermedia (OR 4.03); P. gingivalis and B. forsythus (OR 3.84); P. gingivalis and T. denticola (OR 4.17); P. gingivalis, B. forsythus, and T. denticola (OR 4.06); and P. gingivalis, P. nigrescens, and T. denticola (OR 3.29). EBV-1 also showed positive association with severe periodontitis (OR 5.09), with increasing age (OR 1.03), and with periodontal probing depth at the sample sites (OR 1.77). HCMV was positively associated with coinfections of P. gingivalis and P. nigrescens (OR 3.23); P. gingivalis, B. forsythus, and P. nigrescens (OR 3.23); and P. gingivalis, P. nigrescens, and T. denticola (OR 2.59); with severe periodontitis (OR 4.65); and with age (OR 1.03). Patients with mixed viral infections revealed significant associations with P. gingivalis (OR 2.27), and with coinfections of P. gingivalis and B. forsythus (OR 2.06); P. gingivalis and P. nigrescens (OR 2.91); P. gingivalis, B. forsythus, and P. nigrescens (OR 2.91); and P. gingivalis, P. nigrescens, and T. denticola (OR 2.70) with the clinical diagnosis of slight (OR 3.73), moderate (OR 3.82), or severe periodontitis (OR 4.36), and with probing depth at the sample sites (OR 1.39). HSV and EBV-2 showed no significant associations with any of the variables tested. CONCLUSIONS: The results indicate that subgingival EBV-1, HCMV, and viral coinfections are associated with the subgingival presence of some periodontal pathogens and periodontitis. Herpesviruses may exert periodontopathic potential by decreasing the host resistance against subgingival colonization and multiplication of periodontal pathogens. PMID- 10368052 TI - The relationship between oral malodor, gingivitis, and periodontitis. A review. AB - Volatile sulfur compounds (VSC) are a family of gases which are primarily responsible for halitosis, a condition in which objectionable odors are present in mouth air. Although most patients perceive this condition as primarily a cosmetic problem, an increasing volume of evidence is demonstrating that extremely low concentrations of many of these compounds are highly toxic to tissues. VSC may, therefore, play a role in the pathogenesis of inflammatory conditions such as periodontitis. Since these compounds result from bacterial putrefaction of protein, investigations have been conducted to determine whether specific bacteria are associated with odor production. Two members of this family, hydrogen sulfide (H2S) and methyl mercaptan (CH3SH), are primarily responsible for mouth odor. Although many bacteria produce H2S, the production of CH3SH, especially at high levels, is primarily restricted to periodontal pathogens. Direct exposure to either of these metabolites adversely affects protein synthesis by human gingival fibroblasts in culture. However, methyl mercaptan has the greatest effect. Other in vitro experiments have demonstrated that cells exposed to methyl mercaptan synthesize less collagen, degrade more collagen, and accumulate collagen precursors which are poorly cross-linked and susceptible to proteolysis. CH3SH also increases permeability of intact mucosa and stimulates production of cytokines which have been associated with periodontal disease. VSC, and in particular methyl mercaptan, are therefore capable of inducing deleterious changes in both the extracellular matrix and the local immune response of periodontal tissues to plaque antigens. This article reviews these data and emphasizes the potential importance of VSC in the transition of periodontal tissues from clinical health to gingivitis and then to periodontitis. PMID- 10368053 TI - Two multi-center studies evaluating locally delivered doxycycline hyclate, placebo control, oral hygiene, and scaling and root planing in the treatment of periodontitis. AB - BACKGROUND: The clinical efficacy and safety of doxycycline hyclate (8.5% w/w) delivered subgingivally in a biodegradable polymer (DH) was compared to placebo control (VC), oral hygiene (OH), and scaling and root planing (SRP) in 2 multi center studies. METHODS: Each study entered 411 patients who demonstrated moderate to severe periodontitis. Patients had 2 or more quadrants each with a minimum of 4 qualifying pockets > or =5 mm that bled on probing. At least 2 of the pockets were > or =7 mm. Treatment with DH, VC, OH, or SRP was provided at baseline and again at month 4. Clinical parameters were recorded monthly. RESULTS: DH and SRP resulted in nearly identical clinical changes over time in both studies. Mean 9 month clinical attachment level gain (ALG) was 0.8 mm for the DH group and 0.7 mm for the SRP group in Study 1, and 0.8 mm (DH) and 0.9 mm (SRP) in Study 2. Mean probing depth (PD) reduction was 1.1 mm for the DH group and 0.9 mm for the SRP group in Study 1 and 1.3 mm for both groups in Study 2. Frequency distributions showed an ALG > or =2 mm in 29% of DH sites versus 27% of SRP sites in Study 1 and 31% of DH sites versus 34% of SRP sites in Study 2. PD reductions > or =2 mm were seen in 32% of DH sites versus 31% of SRP sites in Study 1 and 41% of DH sites versus 43% of SRP sites in Study 2. Comparisons between DH, VC, and OH treatment groups showed DH treatment to be statistically superior to VC and OH. Safety data demonstrated a benign safety profile with use of the DH product. CONCLUSIONS: Results of this trial demonstrate that treatment of periodontitis with subgingivally delivered doxycycline in a biodegradable polymer is equally effective as scaling and root planing and superior in effect to placebo control and oral hygiene in reducing the clinical signs of adult periodontitis over a 9-month period. This represents positive changes resulting from the use of subgingivally applied doxycycline as scaling and root planing was not limited regarding time of the procedure or use of local anesthesia. PMID- 10368054 TI - Persistence of oral colonization by the same Actinobacillus actinomycetemcomitans strain(s). AB - BACKGROUND: The Gram-negative facultatively anaerobic coccobacillus Actinobacillus actinomycetemcomitans is the major pathogen in localized juvenile periodontitis (LJP) and some forms of adult periodontitis (AP). A. actinomycetemcomitans can be grouped into 5 serotypes (a through e) based on differences in the carbohydrate moiety of cell surface lipopolysaccharide. The A. actinomycetemcomitans population is genetically heterogeneous. Since the studies on A. actinomycetemcomitans colonization have mostly applied only culture techniques, the clonality of the follow-up isolates has not been established. Thus, it is possible that, although A. actinomycetemcomitans could be repeatedly isolated from an individual, the initial colonizing strain was replaced by another strain. The aim of the study was to determine whether oral A. actinomycetemcomitans strains change spontaneously over time or after periodontal treatment. METHODS: A total of 922 A. actinomycetemcomitans isolates were recovered from 115 subjects. From each subject A. actinomycetemcomitans isolates were obtained from 2 to 9 follow-up samples 0.5 to 11.5 years apart. After the first sampling occasion, 99 subjects were treated for either LJP or AP, whereas the 16 non-periodontitis subjects received no treatment. All A. actinomycetemcomitans isolates were serotyped and 235 isolates from 52 subjects genotyped with AP-PCR and/or with ribotyping. RESULTS: Isolates of only one serotype, or non-serotypeable isolates alone, were repeatedly found in 104 subjects; serotype a occurred in 25%, b in 33%, c in 23%, d in 5%, e in 7%, and non-serotypeable isolates in 8% of these subjects. Two serotypes (or serotypeable isolates together with non-serotypeable isolates) occurred simultaneously in 9 subjects and in each of these subjects at least one of the serotypes was detected at each sampling occasion. In one subject the initial serotype reappeared although a different serotype was once seen alone, whereas in another subject the initial serotype could not be recovered later. Identical genotypes of A. actinomycetemcomitans were repeatedly detected in each of 52 subjects with follow up isolates of the same serotype. CONCLUSIONS: The results showed that spontaneous or treatment-induced change in the oral A. actinomycetemcomitans strain(s) is extremely rare and that colonization with the same strain(s) seems to be remarkably persistent. PMID- 10368055 TI - The use of 2 bioabsorbable barrier membranes in the treatment of interproximal intrabony periodontal defects. AB - BACKGROUND: The use of barrier membranes in the treatment of periodontal defects is well documented. There has been an increase in the use of bioabsorbable materials which do not require a second surgical procedure for removal. However, there are little data evaluating the efficacy of bioabsorbable membranes in the treatment of intrabony defects. The purpose of this investigation was to evaluate the regenerative potential of 2 bioabsorbable barrier membranes without the use of grafting materials in the treatment of interdental intrabony defects. METHODS: Twenty-three 2- or 3-walled intrabony defects were treated in 19 patients with a mean age of 50.4 years. All had completed nonsurgical treatment and a period of supportive periodontal therapy. The sites were randomly chosen to receive a barrier membrane composed of type I bovine collagen (11) or a copolymer of polylactic acid (PGA/PLA;12). A pressure sensitive disc probe was used to evaluate the following criteria at baseline and re-entry: 1) occlusal surface to the apical depth of probe penetration (OS-DP); 2) occlusal surface to the gingival margin (OS-GM); 3) occlusal surface to the alveolar crest (OS-AC); and 4) occlusal surface to the base of the osseous defect (OS-BD). Full thickness mucoperiosteal flaps were reflected to expose the surgical sites. The defects were debrided of the granulomatous tissue, the root surfaces instrumented and conditioned with 4 one-minute applications of 50 mg/ml of tetracycline. The barrier membranes were adapted to cover the defects and the flaps replaced. The postsurgical healing was uneventful and similar in both treatment modalities. RESULTS: Twenty-three sites were surgically re-entered 6 months from the time of the initial surgery. The deepest probe depth for each site was used for statistical analysis. There was a mean relative attachment gain of 2.58+/-1.90 mm for the collagen, and 2.77+/-2.13 mm for the copolymer. There was a decrease in probing depth of 3.27+/-1.91 mm and 0.69+/-1.35 mm of recession for the collagen. The PGA/PLA copolymer had 3.55+/-2.47 mm reduction in probe depth and 0.78+/-1.14 mm of recession. CONCLUSIONS: The data indicated the bioabsorbable collagen and copolymer membranes resulted in comparable results. A larger sample size would be necessary to determine if one membrane was superior to the other. PMID- 10368057 TI - Comparison of bioactive glass to demineralized freeze-dried bone allograft in the treatment of intrabony defects around implants in the canine mandible. AB - BACKGROUND: The purpose of this study was to evaluate and compare the healing of different bone grafting materials adjacent to titanium plasma-sprayed (TPS) endosseous dental implants. METHODS: Implant osteotomy sites were prepared and standardized 3-walled intrabony defects (3 mm x 5 mm x 5 mm) were created at the mesial of each implant site. Thirty-two TPS implants were placed in edentulous mandibular ridges of the 4 dogs. Periodontal dressings were placed in the defect sites so as to create a defect simulating bone loss around an implant. After 3 months, the periodontal dressing was removed, the defect sites debrided and evaluated for size, and intramarrow penetration performed. The graft materials tested were 1) canine demineralized freeze-dried bone allograft (cDFDBA); 2) bioactive glass granules of a broad size range 90 to 710 microns (BRG); and 3) bioactive glass granules of narrow size range 300 to 355 microns (NRG). One site on each side of the mandible was not filled and served as a control. Dogs were sacrificed 4 months after graft placement. RESULTS: Histologically, differences in percent bone-to-implant contact in the defect area were observed between the treatment groups. cDFDBA>control=BRG=NRG with statistical significance found between cDFDBA and control (P = 0.0379), but no statistically significant difference between control or either bioactive glass material. When comparing percent bone height fill of the defect in the grafted area, cDFDBA (65.7%) was significantly better than the control (48.9%; P < or = 0.05) with no statistically significant difference between control, broad range bioactive glass (57.3%) and narrow range bioactive glass (56.6%). When total bone area was measured, the percentage of new bone in the grafted area was cDFDBA (42.1%), broad range glass (33.1%) and narrow range glass (22.6%) with significance found between cDFDBA and NRG (P = 0.0102). The content of residual graft particles in soft tissue was significant (P = 0.0304) between cDFDBA (1.4%) and NRG (11.4%) with no significant difference between graft material for residual particle content in bone tissue. CONCLUSIONS: The results of this study indicate that percent bone-to-implant contact and percent bone height fill in an intrabony defect around titanium plasma-sprayed implants are statistically significantly higher with the use of DFDBA when compared to bioactive glass material. PMID- 10368056 TI - Effects of tobacco products on the attachment and growth of periodontal ligament fibroblasts. AB - BACKGROUND: Cigarette smoking is one of the most significant risk factors in the development and further advancement of inflammatory periodontal disease, however, the role of either nicotine or its primary metabolite cotinine in the progression of periodontitis is unclear. This study aimed to investigate the effects of nicotine and cotinine on the attachment and growth of fibroblasts derived from human periodontal ligament (PDL). METHODS: Primary cultures were prepared from the roots of extracted premolar teeth. Cells were used at both low (P3 to P5) and high (P11 to P13) passage. Cell numbers were determined over 14 days using either the 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay or with a Coulter counter. Cultures were exposed to culture medium supplemented with 1) 15% fetal calf serum (FCS) only; 2) 1% FCS only; 3) 1% FCS and nicotine (concentration range 5 ng/ml to 10 mg/ml); or 4) 1% FCS and cotinine (concentration range 0.5 ng/ml to 10 microg/ml). RESULTS: Nicotine significantly (P <0.05, by ANOVA) inhibits attachment and growth of low passage cells at concentrations >1 mg/ml and high passage PDL fibroblasts at concentrations >0.5 mg/ml. Cotinine, at the highest concentration used (10 microg/ml), appeared to inhibit attachment and growth of both low and high passage fibroblasts but this was not statistically significant (P >0.05, by ANOVA). CONCLUSIONS: Tobacco products inhibit attachment and growth of human PDL fibroblasts. This may partly explain the role of these substances in the progression of periodontitis. PMID- 10368058 TI - CREST syndrome and periodontal surgery: a case report. AB - CREST syndrome is a slowly progressive form of systemic scleroderma. It is characterized by calcinosis cutis, Raynaud's phenomenon, esophageal involvement, sclerodactyly, and telangiectasia. There are limited reports of dental treatment for patients with this syndrome, and no reports of periodontal surgical procedures. This paper presents a case report of periodontal surgical treatment in a 38-year-old female patient with CREST syndrome, and a discussion of the clinical manifestations of the syndrome as they relate to dental treatment. PMID- 10368059 TI - Adverse effects associated with a bioabsorbable guided tissue regeneration device in the treatment of human gingival recession defects. A clinicopathologic case report. AB - This clinicopathologic case report documents an adverse effect associated with the use of a polylactic acid-based barrier in the treatment of human gingival recession defects. A total of 27 consecutively treated patients, in whom guided tissue regeneration with a polylactic acid barrier was used to correct gingival recession defects, were evaluated. This adverse effect consisted of a midradicular-apical swelling, generally asymptomatic, with no apparent predilection for gender, age, tooth type or location (maxilla/mandible), or surgical procedure. It was observed in 14 of 27 (52%) patients and 22 of 41 (54%) defects. The swelling decreased in size over time and in most cases, it completely resolved within 12 months postsurgery. Histopathologic evaluation of a 14-week specimen indicated characteristics (multinucleated giant cells, foamy macrophages) consistent with a foreign body reaction. These findings suggest that patients undergoing GTR procedures with synthetic absorbable devices for the treatment of gingival recession defects should be advised of the possible occurrence of such an adverse effect. PMID- 10368060 TI - Effective periodontal treatment in a patient with type IIA von Willebrand's disease: report of a case. AB - von Willebrand's disease (vWD) is one of the most common hereditary hemorrhagic disorders. A mild to moderate deficiency of factor VIII and von Willebrand factor (vWf) often is associated with gingival bleeding. In this case report, the periodontal treatment of a patient with vWD is described. A 45-year-old woman with type IIA vWD was referred for periodontal therapy because of an episode of gingival hemorrhage and percussion pain of teeth #18 and #47. The periodontal findings included probing depths ranging from 2 to 6 mm, horizontal bone loss, and Class II furcation involvement of tooth #46. After consultation with a hematologist, apically positioned flap surgery and hemisection were performed on tooth #46 following completion of oral hygiene instruction, scaling and root planing, and endodontic therapy. The patient was given 500 units of factor VIII including vWf multimer 30 minutes before surgery. After healing of the periodontal tissue, prosthodontic treatment was undertaken on the posterior mandibular sextants. At follow-up, the probing depths ranged from 2 to 3 mm, and gingival bleeding on probing was minimal. The patient's children all had vWD. They had mild to moderate periodontitis with probing depths ranging from 2 to 5 mm and gingival bleeding on probing. With the combined efforts of the periodontist and hematologist, effective periodontal treatment can be provided to patients with von Willebrand's disease. PMID- 10368061 TI - Polymer-assisted regenerative therapy: case reports of 22 consecutively treated periodontal defects with a novel combined surgical approach. AB - This report describes the clinical application of an in situ formed barrier of poly(DL-lactide) used in combination with a composite graft of demineralized freeze-dried bone allograft (DFDBA) mixed with calcium sulfate and tetracycline in a ratio of 7:2:1 and citric acid root conditioning for the treatment of intrabony and furcation defects. The clinical outcome was assessed by changes in clinical attachment level (CAL) and probing depth (PD) in 18 consecutively treated patients with 17 intrabony and 5 furcation lesions. After patients demonstrated acceptable oral hygiene, the lesions were surgically treated with combination therapy using an in situ formed barrier over a DFDBA composite graft. Patients followed a stringent postoperative protocol and were evaluated at 6 months postsurgery. CAL improved for all sites from a presurgical average of 8.8+/-2.3 mm to 4.4+/-1.6 mm at 6 months postsurgery (4.4+/-1.5 mm gain), while PD was reduced from an average of 8.3+/-2.1 mm presurgery to 3.3+/-1.1 mm at 6 months postsurgery (5.0+/-1.8 mm reduction). Five furcations were treated, of which 4 were Class II and 1 was Class III. Of these furcation lesions, 3 had complete clinical closure, while 1 improved by 1 grade. The Class III furcation remained the same. Results suggest that DFDBA composite graft covered by an in situ formed barrier on root surfaces treated with citric acid can enhance the prognoses of teeth with periodontal lesions as measured by CAL gains and PD reductions. Further studies are warranted to compare this treatment to other more traditional forms of regenerative therapy to determine its comparative efficacy. PMID- 10368062 TI - Re: Ability of commercial demineralized freeze-dried bone allograft to induce new bone formation is dependent on donor age but not gender (1998;69:470-478) PMID- 10368063 TI - Role of the midline brainstem in feline amygdaloid kindling. AB - PURPOSE: To study the role played by the midline brainstem in feline amygdaloid kindling in general and the formation of transhemispheric positive transfer effect (PTE) in particular. METHODS: Eight adult male cats with either anterior or posterior midline brainstem bisection underwent primary and secondary site kindling and primary site retest. RESULTS: Stage 4 seizure pattern was altered with either absence of contralateral circling in the anterior group or emergence of ipsilateral circling in the posterior group. Electroclinical manifestation of the tonic phase was absent in the posterior but not the anterior group. Kindled convulsive pattern remained asymmetrical with contralateral dominance in the anterior but not the posterior group. Intermittent truncation of the secondary clonic phase of the kindled seizure occurred in both groups. PTE was reduced whereas the negative transfer effect was preserved in both groups. CONCLUSIONS: The anterior half of the midline brainstem participates in stage 4 contralateral circling. Both the anterior and posterior halves of the midline brainstem participate in the mechanisms of repetitive circling during stage 4 and the stable maintenance of the kindled stage 6 seizure. The anterior and posterior halves of the midline brainstem participate in symmetrical patterning and electroclinical tonic expression of the kindled seizure, respectively. The midline brainstem, extending from the midbrain to the pons with some posterior emphasis, participates in the mechanisms of PTE formation. The findings indicate the importance of the brainstem for electroclinical ictal processes and a transhemispheric positive transfer effect associated with temporal limbic epileptogenesis. PMID- 10368064 TI - FOS induction in brain associated with seizure and sustained cortical vasodilation in anesthetized rat. AB - PURPOSE: By estimating the anatomical distribution of neurons expressing c-fos protein, we sought to establish whether the intrinsic neural systems known to be implicated in the cerebrovascular regulation were activated during the increase in cortical blood flow associated with epileptic seizures. METHODS: A single unilateral microinjection of the cholinergic agonist, carbachol, in the thalamic generalized convulsive seizure area was used in anesthetized rats to elicit recurrent episodes of electrocortical epileptiform activity and an increase in cortical blood flow. Neuronal expression of Fos protein was analyzed to identify activated brain regions. RESULTS: We identified two cortical vasodilatory responses: a sustained cortical vasodilatory response associated with the continuous low-frequency, high-amplitude spiking and a transient cortical vasodilatory response invariably related to the recurrent spike-burst activity. The sustained cortical blood flow began to increase at 55-65 min, remaining significantly (p < 0.05) increased and reaching at the end of the experiment < or =182+/-17% of the prestimulated control. The electrocortical epileptic activity and the cerebral cortical vasodilation were associated with a marked increase in Fos immunoreactivity in the entorhinal and piriform cortices, the dentate gyrus, the hippocampus, and the amygdala. Fos-positive neurons also were found in specific thalamic nuclei, the cerebral cortex, the caudate-putamen, the hypothalamus, the pontine parabrachial nuclei, the dorsal raphe, and the rostral ventrolateral medulla. CONCLUSIONS: These results provide evidence that convulsive seizures elicited by cholinergic stimulation of the thalamus, in addition to limbic and somatic motor systems, activate central autonomic nuclei and their pathways, including those implicated in cerebrovascular regulation. PMID- 10368065 TI - Localization of the epileptogenic zone by ictal and interictal SPECT with 99mTc ethyl cysteinate dimer in patients with medically refractory epilepsy. AB - PURPOSE: To evaluate the accuracy, feasibility and clinical value of both ictal and interictal 99mTc-ethyl cysteinate dimer (ECD) single photon emission computed tomography (SPECT) in patients with medically refractory epilepsy. METHODS: The study included 75 consecutive patients, 48 with temporal lobe epilepsy (TLE group), and 27 with extratemporal epilepsy (ExT group). The accuracy of SPECT was analyzed considering the final diagnosis reached by convergence of clinical, electrophysiologic, structural, pathologic and outcome data. RESULTS: Ictal SPECT correctly identified the epileptogenic zone in 21 (91.3%) of 23 patients, whereas interictal SPECTs could correctly identify the epileptogenic zone in only 41 (62.1%) of 66 patients (chi2 = 5.56, df = 1, p < 0.05). Results were similar when the two study groups were analyzed separately. Moreover, ictal studies had significantly higher specificity (91.3 vs. 60.6%) and positive predictive value (91.3 vs. 66.2%) than interictal studies for the whole series of patients. Considering all tools used in the preoperative workup of these patients, ictal SPECT significantly contributed to the final topographic diagnosis in seven of 14 patients from TLE group and in six of nine patients from the ExT group. In these patients, ictal SPECT either obviated the need for invasive EEG or helped to define where to concentrate the efforts of invasive investigation. CONCLUSIONS: These data demonstrate that ictal SPECT can be easily achieved by using 99mTc-ECD and can accurately localize the epileptogenic zone in both temporal and extratemporal epilepsies. Ictal ECD SPECT proved to be significantly more sensitive and specific than interictal ECD SPECT, and clinically useful in the definition of the epileptogenic zone. PMID- 10368066 TI - Ketosis and epilepsy: 31P spectroscopic imaging at 4.1 T. AB - PURPOSE: To determine whether changes in the high-energy phosphates occur with use of the ketogenic diet in patients with intractable epilepsy. METHODS: 31P magnetic resonance spectroscopic imaging studies were performed at 4.1 T in seven patients with intractable epilepsy (four Lennox-Gastaut syndrome, one absence, one primary generalized tonic-clonic, and one partial complex) before and after institution of the ketogenic diet. Coronal 1H anatomic imaging also was performed to provide correlation to the 31P data. RESULTS: Taking the patients as a group, the ratio of phosphocreatine (PC)/gamma-adenosine triphosphate (ATP) measured at baseline (regular diet) compared with that measured after the ketogenic diet showed a small but significant increase from 0.61+/-0.08 to 0.69+/-0.08 (p < 0.05). Comparing the ratio of PCr inorganic phosphorus (Pi) measured at baseline with the postketogenic diet, there was a significant increase from 2.45+/-0.27 to 2.99+/-0.44 (p < 0.05). CONCLUSIONS: As a group, improvement of energy metabolism occurs with use of the ketogenic diet. This is in agreement with the chronic ketosis studies performed earlier in rodents. PMID- 10368067 TI - Differential neuronal and glial relations with parameters of ictal discharge in mesial temporal lobe epilepsy. AB - PURPOSE: EEG recordings of spontaneous seizures should provide clues to epilepsy pathogenesis and pathology. We examined onset, propagation, and termination characteristics of spontaneous seizures recorded by intracranial EEG in temporal lobe (TL) epilepsy in relation to neuronal, glial, and synaptic changes in the same tissue. METHODS: All of our patients with intracranial EEG recordings of spontaneous TL seizure onset, subsequent TL resection, and quantitative pathologic analysis of resected tissue were included. Seizure parameters were mean time to initial propagation, mean total electrical duration, uniformity of seizure-onset distribution and location, and percentage of seizures with spiking onset per patient. Tissue was analyzed for glial and neuronal density in hippocampal fields and presence of sprouting. Outcome was classified as seizure free or not. RESULTS: All seizures with onset in resected TL in 62 patients were analyzed. The percentage of each patient's seizures with spike onsets was significantly correlated with glial density in CA3 (p < 0.01). Initial propagation time was significantly and inversely correlated with neuronal density in CA4 (p < 0.02). Electrical seizure duration was significantly correlated with glial density in CA2 and CA3 (p < 0.02). Neuronal and glial density were significantly (and inversely) related to one another only in CAI (p < 0.001). Outcome was most significantly related to uniform hippocampal seizure onset. Presence or absence of sprouting was not significantly related to outcome or any EEG measure. CONCLUSIONS: These results suggest both glia and neurons exert independent influences on the expression of ictal discharges in seizures of medial TL onset. Glial density influenced interictal-ictal transition, whereas neuronal density influenced seizure propagation. These findings may have implications for pathogenesis. PMID- 10368068 TI - Kinetics of serum neuron-specific enolase and prolactin in patients after single epileptic seizures. AB - PURPOSE: To investigate and compare the temporal profile of serial levels of neuron-specific enolase (NSE) and prolactin in serum from patients after single epileptic seizures. METHODS: Measurement of NSE and prolactin by sensitive immunoassays in 21 patients with complex partial seizure (CPSs: n = 11) and secondarily generalized tonic-clonic seizures (SGTCSs; n = 10) during continuous video-EEG monitoring at four different time points (1, 3, 6, and 24 h after ictal event). Statistical analysis was performed by using a repeated-measures analysis of variance (ANOVA) model. RESULTS: Mean+/-SD values for NSE levels (ng/ml) were 12.5 +/-4.4 (1 h), 10.8+/-3.8 (3 h), 11.1+/-4.9 (6 h), and 8.2+/-1.9 (24 h). The corresponding prolactin levels (mU/L) were 1,311+/-1,034, 232+/-158, 237+/-175, and 251+/-98. There was a significant decrease of NSE and prolactin levels over time (p < 0.001). The pair-wise comparison of NSE levels showed significant differences between the time points 1 vs. 24 h (p < 0.001), 3 vs. 24 h (p = 0.007), and 6 vs. 24 h (p = 0.009). In contrast, serum prolactin levels showed a significant difference between 1 vs. 3 h (p < 0.001) only. Most of the NSE levels remained normal after CPSs and SGTCSs. At 1 h after the seizure, only 33% of the subjects had increased NSE, whereas abnormal prolactin levels occurred with a sensitivity of 80%. CONCLUSIONS: In contrast to prolactin, serum NSE is not a sensitive marker of individual seizures. Only some individuals showed an increase of NSE beyond the prolactin-sensitive time frame after a single seizure, and mean NSE levels were not significantly increased compared with those of normal controls. PMID- 10368069 TI - Risk factors for a first febrile convulsion in children: a population study in southern Taiwan. AB - PURPOSE: To identify risk factors for a first febrile convulsion among 3-year-old children by a matched case-control population study. METHODS: All 11,714 neonatal survivors born in Tainan City between October 1989 and September 1991 were enrolled. At age 3, 10,460 children were available for telephone survey for febrile convulsions, and were confirmed by home visit interviews. Those without history of seizure were randomly matched to each febrile convulsion case by age, gender, and residence district. RESULTS: Two hundred fifty six children had febrile convulsions, and 218 of them and their matched controls were available for analysis. The febrile convulsion cases had significantly more febrile episodes (four or more) per year (33.0 vs. 22.5%; p = 0.021), and cases had a higher percentage of developmental delay (3.7 vs. 0.4%; p = 0.046) and a higher percentage of febrile convulsions in their siblings (12 vs. 0.4%; p = 0.011) than controls. The other sociodemographic, environmental, and biologic variables showed no differences between cases and controls. Step-wise logistic regression showed a highly significant independent association between febrile convulsions and history of febrile convulsions in the siblings, and a moderate one between febrile convulsions and the number of febrile episodes per year. CONCLUSIONS: The presence of febrile convulsions in the siblings and the number of fever episodes per year were the independent and significant predictors of febrile convulsion for an individual case in our population-based sample. PMID- 10368070 TI - The early prognosis of epilepsy in childhood: the prediction of a poor outcome. The Dutch study of epilepsy in childhood. AB - PURPOSE: To examine which variables available early in the course of childhood epilepsy are associated with a poor short-term outcome and to develop models to predict such an outcome. METHODS: We prospectively followed up 466 children with newly diagnosed epilepsy for 2 years. Variables were collected at intake and after 6 months. Outcome was defined as the duration of the terminal remission (TR): poor (<6 months) and not poor (> or =6 months). RESULTS: Of the subjects, 31% had a poor outcome. Multivariate analysis based on the intake variables identified number of seizures, seizure type, and etiology as risk factors for a poor outcome. With the intake and 6-month variables combined, seizure type, etiology, the number of seizures, and not attaining a 3-month remission during these 6 months, and the EEG at 6 months were predictive variables. A predictive model based on the multivariate logistic-regression analysis with the intake variables was correct in 56% of the children in whom it predicted a poor outcome and in 73% of the children in whom it predicted a not-poor outcome. With the intake and 6-month variables together, these percentages were 66 and 79%, respectively. The sensitivity of these models was low (29 and 47%, respectively); the specificity was good (90 and 89%). CONCLUSIONS: The 2-year outcome of childhood epilepsy is closely related to its early course. The prognosis is poor in approximately 30% of patients. By using our data, the prediction of a poor outcome is correct in almost two thirds of the patients; however, the models produce many false-negative predictions. PMID- 10368071 TI - Family interactions as targets for intervention to improve social adjustment after epilepsy surgery. AB - PURPOSE: To identify family interactions associated with psychosocial outcome of epilepsy surgery, to design interventions to improve patient outcome. METHODS: A cross-sectional, case series study of relations among observed family behavior and psychosocial outcome of 43 patients after temporal lobectomy. Videotaped family behavior during family discussion tasks was rated for predominant family affect, affective range, and support of patient autonomy. Multiple regression analyses tested the relation of observed family characteristics to outcomes, controlling for seizure control and other psychological and disease characteristics. RESULTS: Predominant family affect predicted patients' social adjustment independent of postoperative seizure status and other disease characteristics. The relation between predominant affect and social adjustment was stronger among patients with persisting complex partial seizures (CPSs; r = 0.91), versus patients with auras (r = -0.38) and seizure-free patients (r = 0.28; multiple R = 0.71; p < 0.05). Families with a positive affective climate supported patients' autonomy. CONCLUSIONS: Two potential targets were identified for family intervention to improve postsurgical social adjustment: (a) family interactions that support a predominantly positive affective climate, and (b) family interactions that support patient autonomy. These findings are consistent with findings in normal and other clinical populations. They identify specific interactions that give rise to positive versus negative affective climate and support versus undermining of autonomy. These results lay the groundwork for intervention studies targeting these specific family interactions. Such intervention studies would clarify the direction of effect of the observed relationships and would test the efficacy of family intervention for improving psychosocial outcomes for patients with epilepsy. PMID- 10368072 TI - Risk of epilepsy in patients with multiple sclerosis: a population-based study in Iceland. AB - PURPOSE: Several clinical series reported an association between multiple sclerosis (MS) and epilepsy. We conducted a total population study in Iceland to determine the risk for developing epilepsy in patients with MS compared with that expected in the general population. METHODS: Medical records of the 188 incidence cases of clinically definite MS first diagnosed in Iceland during the 25-year study period (1965-1989) were reviewed. The cases were followed up through 1994 or until death to identify those developing seizures or epilepsy. The expected number of cases with epilepsy in the MS-incidence cohort were calculated based on the age-specific incidence for epilepsy in Iceland and the age-specific person years of follow-up in the MS cohort. RESULTS: During the 2,771 person years of observation after diagnosis of clinically definite MS, three MS patients developed epilepsy. One additional case developed epilepsy after onset of MS symptoms but before diagnosis of MS. The cumulative incidence of epilepsy by 10 years after diagnosis of MS was 1.9%. Given the age-specific person years of follow-up after diagnosis of MS, only one case of epilepsy would have been expected; standardized incidence ratio (SIR), 3.0 (95% confidence interval (CI), 0.6-8.8). CONCLUSIONS: MS is a risk factor for developing epilepsy. Patients with MS have a threefold increase in risk for developing epilepsy when compared with that expected in the general population. The reason for this increased risk is unclear and needs further investigation. PMID- 10368073 TI - The descriptive epidemiology of infantile spasms among Atlanta children. AB - PURPOSE: To determine the population-based epidemiology of infantile spasms (IS) among Atlanta children. METHODS: By using data from a cross-sectional, population based surveillance system that included 21 EEG laboratories, we identified children born in 1975-1977 in metropolitan Atlanta with IS. Cumulative incidence up to age 2 years was estimated from the number of children with IS born in the study area in 1975-1977, and age-specific prevalence was calculated from the number of children previously diagnosed with IS who lived in the study area at age 10 years. Data regarding coexisting disabilities were available from the surveillance system for developmental disabilities. RESULTS: The cumulative incidence of IS was 2.9/10,000 live births; half of the children with IS had cryptogenic IS. The age-specific prevalence of IS was 2.0/10,000 among 10-year old children. Eighty-three percent of 10-year-old children with a history of IS had mental retardation (MR, IQ < or =70); 56% of children with a history of IS had profound MR (IQ <20). Developmental outcome did not differ between the children with cryptogenic IS and those with symptomatic IS. Among the 10-year-old children with profound MR who were living in Atlanta at age 10 years, 12% had a history of IS. Fifty percent of children with IS developed Lennox-Gastaut syndrome (LGS) before age 11 years. CONCLUSIONS: The syndrome of IS is rare in the general population, yet a significant percentage of all children with profound MR and severe childhood epilepsy syndromes in the general population have a history of IS. PMID- 10368074 TI - Prospective population-based study of intermittent and continuous convulsive status epilepticus in Richmond, Virginia. AB - PURPOSE: Previous work suggested that there is a lower mortality for convulsive status epilepticus (SE) with intermittent seizures (intermittent SE) as opposed to SE with continuous seizure activity (continuous SE). A plausible hypothesis to explain this difference is that the shorter ictal time in intermittent SE is responsible for the lower mortality in this group. This study investigates the relative contributions of total ictal time and SE duration to the differing mortalities of intermittent and continuous SE. METHODS: Six hundred forty-five cases of prospectively identified convulsive SE were examined. Nonparametric statistical methods were used to compare continuous SE and intermittent SE variables. Multivariate logistic regression analyses were used to determine which factors were most highly associated with mortality. Intermittent SE cases were analyzed to evaluate the relative contributions of ictal time versus SE duration to mortality. RESULTS: Intermittent SE had a significantly lower mortality than continuous SE (19.6 vs. 31.4%; p < 0.001) in adults but not in children. Intermittent and continuous SE durations did not significantly differ in adult cases but did differ in pediatric cases. Ictal time was significantly shorter than SE duration for intermittent SE in both adults and children. After adjusting for age, etiology, and SE duration, SE type (continuous SE vs. intermittent SE) was shown to have an independent effect on mortality in adults. The relative risk of mortality for continuous SE was 1.79 times that of intermittent SE (p = 0.04). After controlling for SE duration, ictal time did not significantly affect mortality in adults. CONCLUSIONS: Intermittent and continuous convulsive SE were common in both pediatric and adult populations. Intermittent SE had a significantly lower mortality than did continuous SE. This difference in mortality was not completely explained by differences in SE duration, total ictal time, etiology, or age. Further research is needed to identify the factor(s) contributing to the significant difference in mortality between intermittent SE and continuous SE. PMID- 10368075 TI - Depth of EEG suppression and outcome in barbiturate anesthetic treatment for refractory status epilepticus. AB - PURPOSE: Barbiturate anesthetic treatment of patients with refractory status epilepticus (RSE) is often titrated to a burst-suppression record on the EEG. We sought to determine whether the depth of EEG suppression correlated with persistent seizure control in such patients. METHODS: We reviewed the EEGs and clinical course of patients treated with pentobarbital (PTB) for RSE. Persistent seizure control or relapse to status epilepticus after the taper of PTB was determined with reference to the depth of EEG suppression during treatment. RESULTS: Of 35 patients tapering PTB, persistent seizure control was achieved in six of 12 patients reaching a burst-suppression record at greatest depth of EEG suppression and in 17 of 20 patients reaching a "flat" record; three patients with neither pattern had persistent control. Survival also was somewhat better in the more suppressed group. Isolated epileptiform discharges during the barbiturate infusion did not correlate with outcome. Recurrence of electrographic status after PTB taper predicted clinical relapse. CONCLUSIONS: The EEG is important in managing PTB treatment for patients with RSE. Some period of intense seizure and EEG suppression may help in preventing relapse of status after the PTB taper. It is not necessary to suppress all epileptiform discharges, but persistent clinical and EEG monitoring is necessary to avoid relapses. PMID- 10368076 TI - Agreement and correctness in adjusting antiepileptic drug treatment: a need for rational drug treatment? AB - PURPOSE: To study interobserver variation in treatment decisions in a first follow-up contact after initiation of antiepileptic drug (AED) treatment. The results should aid us in the assessment of whether decision support can be of value in this situation. METHODS: Data from patient records were used to construct 270 different test cases containing information about the course of the disease after initiation of drug treatment. The cases were presented to five neurologists from different general hospitals who previously agreed about the diagnosis and the initial treatment for these cases. They were asked to write a prescription for each test case. RESULTS: All five neurologists agreed on a treatment decision in 21.9% of the 265 cases available for analysis. Each neurologist made a decision different from the decisions taken by all other neurologists in 14.0-19.6% of the cases. Kappa values for agreement among individual neurologists as well as for agreement between an individual and the group of his peers were low. In 82.6% of the cases, a majority of the neurologists agreed on a treatment decision. Comparing the decisions of individual neurologists with the majority decision reference (219 cases) showed a significant difference in correctness (range, 67.1-82.6%) among the neurologists. CONCLUSIONS: The fact that a majority decision could be reached in a considerable number of cases, as well as the variability in adjustment of an initiated drug treatment, leads us to the conclusion that decision support can contribute to a rational adjustment of drug treatment. PMID- 10368078 TI - Pharmacokinetics of fosphenytoin in patients with hepatic or renal disease. AB - PURPOSE: The pharmacokinetic behavior of fosphenytoin (FOS), the water-soluble prodrug of phenytoin (PHT), has been characterized in normal subjects. This is the first study of the effect of hepatic or renal disease on the rate and extent of conversion of FOS to PHT. METHODS: A single dose of fosphenytoin (250 mg over a period of 30 min) was administered to subjects with hepatic cirrhosis (n = 4), renal disease requiring maintenance hemodialysis (n = 4), and healthy controls (n = 4). Serial plasma concentrations were measured, and pharmacokinetic parameters were calculated. RESULTS: The mean time to reach the peak plasma FOS concentration was similar for each of the three groups. However, the mean time to achieve peak plasma concentrations of PHT tended to occur earlier in the hepatic or renal disease groups than in healthy subjects. The half-life of FOS was 4.5, 9.2, and 9.5 min for the three groups, respectively. There was a trend toward increased FOS clearance and earlier peak PHT concentration in subjects with hepatic or renal disease. This finding is consistent with decreased binding of FOS to plasma proteins and increased fraction of unbound FOS resulting from decreased plasma protein concentrations associated with these disease states. The conversion of FOS to PHT was equally efficient in subjects with hepatic or renal disease and healthy subjects. CONCLUSIONS: Although the differences in pharmacokinetic parameters between the three groups were not statistically significant, these data suggest the need for close clinical monitoring during FOS administration to patients with hepatic or renal disease. To minimize the incidence of adverse effects in this patient population, FOS may need to be administered at lower doses or infused more slowly. PMID- 10368077 TI - Quantification in patient urine samples of felbamate and three metabolites: acid carbamate and two mercapturic acids. AB - PURPOSE: Previously we proposed and provided evidence for the metabolic pathway of felbamate (FBM), which leads to the reactive metabolite, 3-carbamoyl-2 phenylpropion-aldehyde. This aldehyde carbamate was suggested to be the reactive intermediate in the oxidation of 2-phenyl-1,3-propanediol monocarbamate to the major human metabolite 3-carbamoyl-2-phenylpropionic acid. In addition, the aldehyde carbamate was found to undergo spontaneous elimination to 2 phenylpropenal, commonly known as atropaldehyde. Moreover, atropaldehyde was proposed to play a role in the development of toxicity during FBM therapy. Evidence for atropaldehyde formation in vivo was reported with the identification of modified N-acetyl-cysteine conjugates of atropaldehyde in both human and rat urine after FBM administration. Identification of the atropaldehyde-derived mercapturic acids in urine after FBM administration is consistent with the hypothesis that atropaldehyde is formed in vivo and that it reacts with thiol nucleophiles. Based on the hypothesis that the potential for toxicity will correlate to the amount of atropaldehyde formed, we sought to develop an analytic method that would quantify the amount of relevant metabolites excreted in patient urine. METHODS: We summarize the results of an LC/MS method used to quantify FBM, 3-carbamoyl-2-phenylpropionic acid and two atropaldehyde-derived mercapturic acids in the patient population. RESULTS: Analysis was performed on 31 patients undergoing FBM therapy. The absolute quantities of FBM and three metabolites were measured. CONCLUSIONS: This method demonstrated sufficient precision for the identification of patients exhibiting "abnormal" levels of atropaldehyde conjugates and may hold potential for patient monitoring. PMID- 10368079 TI - Induction of ethinylestradiol and levonorgestrel metabolism by oxcarbazepine in healthy women. AB - PURPOSE: To evaluate the effect of oxcarbazepine (OCBZ) on the pharmacokinetic profile of steroid oral contraceptives. METHODS: Twenty-two healthy women aged 18 44 years were recruited, and 16 of them completed the study. By using a randomized double-blind crossover design, each woman was studied in two different menstrual cycles, during which placebo or OCBZ (maintenance dosage, 1,200 mg/day) was given in randomized sequence for 26 consecutive days with a washout of at least one cycle in between. A steroid oral contraceptive containing 50 microg ethinylestradiol (EE) and 250 microg levonorgestrel (LN) was taken for the first 21 days of each cycle. Plasma concentrations of EE and LN were measured by gas chromatography-mass spectrometry in samples collected at regular intervals on days 21-23 of each cycle. RESULTS: Compared with placebo, areas under the plasma concentration curves (AUC(0-24h, geometric means) decreased by 47% for both EE (from 1,677 to 886 pg.h/ml; p < 0.01) and LN (from 137 to 73 ng.h/ml; p < 0.01), during OCBZ treatment. Peak plasma EE concentrations decreased from 180 pg/ml during the placebo cycle to 117 pg/ml during the OCBZ cycle (p < 0.01), whereas peak plasma LN concentrations decreased from 10.2 to 7.7 ng/ml (p < 0.01). The half-lives of EE and LN also decreased from 13.6 to 7.9 h (p < 0.01) and from 28.8 to 15.8 h, respectively (p < 0.01). CONCLUSIONS: OCBZ reduces plasma concentrations of the estrogen and progestagen components of steroid oral contraceptives, presumably by stimulating their CYP3A-mediated metabolism in the liver or gastrointestinal tract or both. Because this may lead to a decreased efficacy of the contraceptive pill, women treated with OCBZ should receive preferentially a high-dosage contraceptive and should be monitored for signs of reduced hormonal cover. PMID- 10368080 TI - Topiramate pharmacokinetics in infants. AB - PURPOSE: This study's goal was to provide preliminary data on the pharmacokinetics of topiramate (TPM) in a cohort of infants (younger than 4 years) participating in an open-label trial of TPM in refractory infantile spasms. METHODS: The pharmacokinetics of TPM were assessed in infants receiving a stable TPM dose for >7 days during the extension phase of this trial. Blood samples were drawn just before and 0.5. 1, 1.5, 2, 4, 6, 8, and 12 h after the morning TPM dose. TPM plasma concentrations were determined by fluorescence polarization immunoassay. The noncompartmental analysis module of WinNonlin was used to calculate individual patient pharmacokinetics profiles. RESULTS: Five infants (ages, 23.5-29.5 months) formed the study cohort. These infants had been given TPM for a median of 9 months (range, 6-11 months) and were currently receiving between 11 and 38.5 mg/kg/day TPM. One was receiving TPM monotherapy, whereas four were taking concomitant antiepileptic medications (AEDs; n = 2, enzyme-inducing agents; n = 2, non-enzyme-inducing drugs). TPM pharmacokinetics in infants appears to be linear. In this cohort, mean TPM plasma clearance (CL/F, 66.6+/-27.4 ml/h/kg) was slightly higher than that reported for children and adolescents and therefore substantially higher than that reported for adults. TPM CL/F was higher and the calculated half-life shorter in the infants receiving concomitant enzyme-inducing AEDs. CONCLUSIONS: Based on this small cohort of patients, it appears that infants may require significantly larger TPM doses, based on weight, than children, adolescents, or adults. Titration to effect and not absolute TPM dose should guide therapy in this age group. PMID- 10368081 TI - Topiramate and metabolic acidosis. AB - Topiramate (TPM) is a novel antiepileptic medication (AED) with at least three mechanisms of action. A possible fourth mechanism, that of a carbonic anhydrase inhibitor, also may contribute to its antiepileptic properties. We report a patient with intractable epilepsy and normal renal function who developed a normal anion gap metabolic acidosis, which worsened during elective surgery for temporal lobectomy. We believe this side effect of TPM can become clinically significant during surgery, concomitant use of another carbonic anhydrase inhibitor, and potentially with the ketogenic diet. PMID- 10368082 TI - Lamotrigine therapy in juvenile neuronal ceroid lipofuscinosis. AB - PURPOSE: To evaluate the effects of lamotrigine (LTG) therapy on epileptic seizures and general well-being in patients with juvenile neuronal ceroid lipofuscinosis (JNCL). METHODS: LTG was initiated in 28 patients with JNCL. The mean age of the patients at the initiation of LTG was 13.7 years (range, 6.7-28.2 years). LTG was started at a dosage of 0.1-0.5 mg/kg/day and increased every 2 weeks until a maintenance dose of 1.25-15 mg/kg/day was reached. On the basis of the indication for LTG therapy, the patients could be divided into four groups. In the first group, LTG was initiated on an add-on basis; in the second group, LTG was started as the first antiepileptic drug (AED) because of seizures, and in the third group, despite no preceding seizures, because of epileptiform activity in the whole-night polysomnography; in the fourth group, LTG replaced valproate (VPA), which was discontinued because of adverse side effects. The efficacy was assessed after 1 year on LTG. The mean follow-up time was 2.8 years (range, 1.3 5.8). RESULTS: LTG had a favorable effect in 23 of 28 patients. A decrease in frequency of seizures of > or =50% was observed in 10 and a decrease in severity of seizures in nine of the 22 patients who had preceding seizures. Increases in well-being were found in 18 of 28. During the follow-up, LTG was continued as monotherapy in 13 of 19 patients. CONCLUSIONS: In light of our experiences, LTG seems to be a valuable drug in JNCL. PMID- 10368083 TI - Estimation of topiramate in subdural cerebrospinal fluid, subcutaneous extracellular fluid, and plasma: a single case microdialysis study. AB - PURPOSE: To estimate topiramate (TPM) concentrations in subdural cerebrospinal fluid (CSF), subcutaneous extracellular fluid (ECF), and plasma and to study the correlation of TPM concentrations in these three different compartments. METHODS: In this single case study of a patient with drug-resistant partial epilepsy undergoing presurgical evaluation with subdural EEG monitoring, we used microdialysis to estimate concentrations of unbound TPM in CSF of the subdural space and ECF of abdominal subcutaneous tissue. Blood samples were drawn for estimation of TPM concentrations in plasma. RESULTS: The correlation between unbound TPM concentrations in subdural CSF and abdominal subcutaneous ECF was good. The mean ratio of ECF/CSF TPM concentration was 0.93 (SD+/-0.03) and the correlation coefficient was 0.98. The mean ratio of ECF/total plasma TPM was 0.75 (SD+/-0.06), and the correlation coefficient was 0.99. The mean ratio of CSF/total plasma TPM was 0.81 (SD+/-0.06), and the correlation coefficient was 0.97. CONCLUSIONS: Assuming a protein binding of TPM of approximately 13%. it is concluded that, based on nine microdialysis samples from a single subject, TPM levels in the CSF at the cortical surface are approximately the same as the unbound plasma levels. Additional patients should be studied to confirm the results. PMID- 10368085 TI - Are we ready to get organized? PMID- 10368086 TI - Gustatory sweating: clinical implications and etiologic aspects. AB - PURPOSE: It was the aim of this study to provide detailed general information on the clinical picture of different kinds of gustatory sweating, including reevaluation of a series of patients who underwent parotidectomy, removal of the submandibular gland, or neck dissection. PATIENTS AND METHODS: This study summarizes the statements of 548 patients questioned about the occurrence of gustatory sweating after parotidectomy (n = 296), extirpation of the submandibular gland (n = 79), and neck dissection (n = 173). RESULTS: After parotidectomy, 45% of the patients had noticed gustatory sweating. In most of them (70%), the symptoms began within 6 months after surgery. Gustatory sweating developed in only one patient with submandibular extirpation (1.5%), and not at all after neck dissection. Most patients (52%) reported that the symptoms occurred independent of the kind of food ingested. These results show that the "masticatory component" is an important trigger for Frey's syndrome. Application of Minor's test localized gustatory sweating mainly in the region of previous parotid lobe removal, but also in other areas deriving their sensory supply from the auriculotemporal, greater auricular, and lesser occipital nerves. The size of the area affected by the sweating was similar after lateral and total parotidectomy. When evaluating clinical symptoms, subjective assessment by the patients seemed to play a major role. After submandibular extirpation and neck dissection, some patients reported gustatory sweating that was not verified by Minor's test. CONCLUSION: There is general agreement that the cause of gustatory sweating is sympathetic or parasympathetic innervation of previously denervated sweat glands, initiated by gustatory triggers. The location of the "erroneous innervation" depends on the type of lesion. In cases after parotidectomy, misdirected parasympathetic regeneration is the model integrating all known factors into a rational concept. For didactic and systematic-pragmatic reasons, a clinically oriented classification of gustatory sweating (types I to III) seems to be useful. PMID- 10368087 TI - Factors influencing condylar position after the bilateral sagittal split osteotomy fixed with bicortical screws. AB - PURPOSE: Multiple articles have discussed condylar position after bilateral sagittal split osteotomy (BSSO). However, previous studies have been limited to two-dimensional evaluation of condylar position. The purpose of this study was to evaluate change in condylar position after a BSSO fixed with bicortical screws using three-dimensional computed tomography to assess the factors that may influence the ultimate position of the condyle after surgery. PATIENTS AND METHODS: Seventeen patients underwent isolated mandibular advancement involving a BSSO with rigid fixation. Reformated axial computed tomography was done 1 week before and 8 weeks after surgery. Movements evaluated included 1) medial-lateral, 2) superior-inferior, 3) anterior-posterior, and 4) condylar angulation. Three separate factors were analyzed to study their effects on the four movements noted: 1) amount of mandibular advancement, 2) amount of proximal segment rotation, and 3) preoperative shape of the mandible. A linear regression analysis was used with statistical significance set at P<.05. RESULTS: Eight weeks after a BSSO and mandibular advancement, most cases showed displacement of the condyle medially, posteriorly, superiorly, and angled medially. The amount of mandibular advancement did not correlate with medial-lateral change of the condyle. The amount of advancement correlated with the condyle angulation and superior inferior changes in condyle position. There was no correlation between amount of advancement and medial or anterior-posterior change in condyle position. There also was no correlation between any of the condylar movements and the degree of proximal segment rotation or the shape of the mandible. CONCLUSIONS: There are obvious changes in condyle position after a BSSO. These changes appear to be influenced mainly by factors other than amount of advancement, degree of proximal segment rotation, and shape of the mandible. PMID- 10368088 TI - Root-end cavity preparation after apicoectomy using a new type of sonic and diamond-surfaced retrotip: a 1-year follow-up study. AB - PURPOSE: This study evaluated the outcome of periradicular surgery using a new set of retrotips for root-end cavity preparation. PATIENTS AND METHODS: Forty three patients who had 50 consecutively treated teeth with periradicular pathology were enrolled in this prospective study. After apicoectomy, root-end cavity preparation was performed, using diamond-surfaced retrotips driven by a sonic handpiece; EBA-cement was used as the root-end filling material. Healing assessment was based on clinical and radiographic criteria. RESULTS: At the 1 year follow-up, 82% of the reexamined surgical cases presented with successful healing. Fourteen percent of the treated teeth were deemed as improved (partial healing), and 4% were classified as failures. CONCLUSION: The new retrotips were found to be ideal for root-end cavity preparation. They simplify the surgical approach to root ends where the working space is limited by restricted access. Root-ends prepared with this new sonoabrasive technique yielded excellent results at the 1-year follow-up examination. PMID- 10368089 TI - Accuracy of integration of dental casts in three-dimensional models. AB - PURPOSE: This study investigated errors occurring in three-dimensional (3D) models when plaster dental casts are integrated into them. MATERIALS AND METHODS: Three-dimensional milling models of three patients with a jaw deformity were fabricated using the Endoplan system (SPARC International Inc, Santa Clara, CA). After this, plaster dental casts were integrated into the 3D models using a face bow transfer system. Two cephalograms were then compared, one obtained from the patient and the other obtained from the 3D model painted with contrast medium. RESULTS: In two cases, the reproducibility of the dental position as determined by angle analysis was within 2 degrees, and that determined by distance analysis was within 2 mm. However, errors over 4 degrees and 4.2 mm, respectively, were observed in one case. CONCLUSION: It is clinically important to confirm the accuracy of the 3D model by cephalometric analysis, and it may be necessary to reposition the dental model based on the results. PMID- 10368090 TI - The incidence, location, and height of maxillary sinus septa in the edentulous and dentate maxilla. AB - PURPOSE: This study evaluated the incidence, location, and height of antral septa and demonstrates their clinical implications. MATERIALS AND METHODS: One hundred ninety-four maxillary posterior regions, subdivided into four groups (group 1, 61 clinically examined atrophic ridges; group 2, 41 anatomically examined atrophic ridges; group 3, 42 radiographically [CT] examined atrophic ridges; and group 4, 50 CT examined dentate maxillary ridges), were examined for the incidence, location, and height of antral septa. RESULTS: The incidence of antral septa was significantly greater (P<.01) in atrophic edentulous regions (groups 1, 2, and 3) than in dentate regions (group 4). However, the septa were much lower (P<.01). In atrophic maxillae, about 70% of antral septa were located in the anterior (premolar) region. CONCLUSIONS: Antral septa are more commonly found in edentulous atrophic maxillae than in dentate maxillae. The septae in edentulous atrophic maxillae are shorter than those found in dentate maxillae. When present, maxillary sinus septae are more common anteriorly than posteriorly. CT scanning is the preferred radiographic method for detecting the presence (or absence) of sinus septae. Panoramic radiography has less sensitivity and specificity than CT scanning for the detection of sinus septa. PMID- 10368091 TI - Additive analgesic effects of oxycodone and ibuprofen in the oral surgery model. AB - PURPOSE: A traditional approach to achieve greater analgesic efficacy is to combine an efficacious dose of a nonopioid with a dose of an opioid sufficient to produce additive analgesia without a substantial increase in the incidence of adverse effects. This study evaluated the additive analagesic effects of the combination of ibuprofen and oxycodone. PATIENTS AND METHODS: A dose of 400 mg ibuprofen was compared with 400 mg ibuprofen with oxycodone in doses of 2.5, 5, or 10 mg in the oral surgery model of acute pain. Analgesic efficacy was measured with category and visual analog scales at 15, 30, 45, and 60 minutes and hourly up to 6 hours. RESULTS: Ibuprofen plus 10 mg oxycodone produced significantly greater analgesia compared with the other three groups, as measured by the visual analog scale from 15 minutes after drug administration up to the 2-hour observation. All four treatments were similar from 3 to 6 hours, with the area under the pain intensity difference curve being similar across groups. Neither the 2.5-mg nor the 5-mg oxycodone dose provided any additive analgesia over ibuprofen at any points. Addition of oxycodone resulted in a dose-related increase in the number of patients reporting adverse effects, with significantly greater drowsiness and vomiting at the 10-mg dose. CONCLUSIONS: These results indicate that additive analgesia can be achieved for the combination of a nonsteroidal anti-inflammatory drug and an orally effective opioid, with faster onset of relief for the combination of 400 mg ibuprofen and 10 mg oxycodone over the first 2 hours after administration, but at the expense of an increased incidence of adverse events. PMID- 10368092 TI - Risk factors contributing to symptomatic plate removal in orthognathic surgery patients. AB - PURPOSE: This study analyzed the fate of miniplates in orthognathic surgery and defined risk factors that eventually result in plate removal. PATIENTS AND METHODS: The outpatient clinic files of 70 patients who had undergone orthognathic surgery were reviewed. All osteotomies were rigidly fixed with stainless steel or titanium miniplates. Study variables included age, gender, plate material, site of plates, and reasons for plate removal. RESULTS: Of 260 plates used for fixation, 31 were removed (12%). When all factors were considered together, only age was statistically significant. Patients older than 30 years of age were more likely to have plate removal (22% vs. 9%). Only when each factor was considered separately were gender and plate material statistically significant. Females (15.4% vs. 6.7%) and stainless steel plates (15.5% vs. 6.7%) were more prone to plate removal. Although more plates were removed from the buttress (15.5%) and chin (14.5%) compared with the piriform area (6.4%), this was not statistically significant. CONCLUSIONS: Age can be defined as a primary risk factor for plate removal, whereas gender and plate material are secondary. Although age and gender are not controllable, the use of titanium plates and infection control may lower the number of symptomatic plates and the need for their removal. PMID- 10368093 TI - Identification of Chlamydia trachomatis in the human temporomandibular joint. AB - PURPOSE: Reactive arthritis (ReA) as a consequence of triggering Chlamydia trachomatis infections has been extensively studied to better understand inflammatory arthritis. This study investigated whether the presence of C trachomatis can be shown in the TMJ of patients with internal derangement. PATIENTS AND METHODS: Posterior bilaminar tissue removed from 31 patients (29 F, 2 M) during TMJ articular disc repositioning and posterior ligament repair was tested for the presence of C trachomatis. Cryosections were stained using a monoclonal antibody that identifies all chlamydial serovars. Highly specific polymerase chain reaction (PCR) assays independently targeting two genes of C trachomatis also were performed; these assays also identify all serovars of this organism. RESULTS: TMJ tissue from 6 of 30 patients (20%) showed the presence of C trachomatis in the posterior bilaminar tissue on immunostaining. PCR screening identified 12 of 31 patients (39%) as having C trachomatis DNA in tissue, including four of six positive by immunostaining. All chlamydia-positive patients were female, with an average age of 36.7 years (15 to 48 years). CONCLUSIONS: The presence of C trachomatis in the human TMJ has not been previously shown. The presence of this organism may serve as the pathogenetic mechanism for TMJ dysfunction, as demonstrated in other joints. Nonapparent chlamydial infection in females may also explain the marked prevalence of TMJ symptoms in women. PMID- 10368094 TI - Three-dimensional computed tomography landmark measurement in craniofacial surgical planning: experimental validation in vitro. AB - PURPOSE: This study evaluated the measurement accuracy of three-dimensional (3D) volumetric images from spiral computed tomography (CT) in vitro. MATERIALS AND METHODS: The study sample consisted of nine cadaver heads that were submitted to an impact force by a special device to promote blunt traumatic craniofacial fractures. The heads were subsequently scanned by a spiral CT scanner (Toshiba Xpress S/X). The archived CT data were transferred to networked computer workstations (Sun Microsystems with Cemax VIP version 1.4 software) to generate 3D volumetric images. The visualization software was used to make interactive linear measurements on the 3D images. Measurements were made on the images twice by two observers, based on conventional craniofacial anatomic landmarks. The soft tissues were subsequently removed, and the same measurements were repeated on the cadaver heads with an electromagnetic digitizer (3 Space, Polhemus, Colchester, VT). RESULTS: The results showed no statistically significant differences between the 3D-CT and the physical measurements, with P>.05 for all measurements. The mean difference between the image and real measurements was less than 2 mm in all instances. CONCLUSIONS: It is concluded that measurement of the skull and facial bone landmarks by 3D reconstruction is quantitatively accurate for surgical planning and treatment evaluation of craniofacial fractures. PMID- 10368096 TI - Mechanical properties of trabecular bone in the human mandible: implications for dental implant treatment planning and surgical placement. AB - PURPOSE: This study sought to establish the relationships between bone density, elastic modulus, and ultimate compressive strength of trabecular bone in the human mandible, and to determine the influence that the cortical plates have on these values. MATERIALS AND METHODS: Nine fresh-frozen human mandibles between the ages of 56 and 90 years were cut into anterior (incisors and canine), middle (premolars), and distal (molars) sections. Seventy-six cylindrical trabecular bone specimens with bone marrow in situ were then prepared and tested in compression in the vertical direction. These tests were performed at a constant strain rate of 0.01 s(-1) with and without the presence of the cortical plates. RESULT: The density of mandibular trabecular specimens with bone marrow in situ ranged from 0.85 to 1.53 g/cm3, with a mean value of 1.14 g/cm3 (SD = 0.15). With the cortical plates present, the elastic modulus ranged from 24.9 to 240.0 megapascals (MPa), with a mean value of 96.2 MPa (standard deviation (SD) = 40.6). Without the cortical plates present, the elastic modulus ranged from 3.5 to 125.6 MPa, with a mean value of 56.0 MPa (SD = 29.6). The ultimate compressive strength of the trabecular bone ranged from 0.22 to 10.44 MPa, with a mean value of 3.9 MPa (SD = 2.7). CONCLUSION: This study indicates that the trabecular bone in the human mandible possesses significantly higher density, elastic modulus, and ultimate compressive strength in the anterior region than in either the middle or distal regions. The absence of cortical plates decreases the bone elastic modulus. These findings quantitatively confirm the need for clinical awareness in altering implant treatment plans and/or design in relation to bone density and the presence of the cortical plates. PMID- 10368095 TI - Bone regeneration produced in rat femur defects by polymer capsules containing recombinant human bone morphogenetic protein-2. AB - PURPOSE: This study examines the effect of polymer capsules containing recombinant human bone morphogenetic protein-2 (rhBMP-2) on bone regeneration in a large segmental bone defect in the rat. MATERIALS AND METHODS: Poly(DL-lactide co-glycolide) (PLGA) capsules containing 3 microg of rhBMP-2 were implanted in a 5-mm segmental femoral defect in rats, and the femurs were harvested at 4 and 8 weeks after implantation and observed by radiographically and microscopically. RESULTS: At 8 weeks after implantation, union of bone was found radiographically and histologically in the femoral defects implanted with the rhBMP-2/PLGA capsules. In contrast, the control animals did not show bridging of the defect. CONCLUSIONS: This study provides evidence that use of rhBMP-2/PLGA capsules is a promising delivery system to regenerate bone. PMID- 10368097 TI - Radiopaque mass in the submandibular region. PMID- 10368098 TI - Basic science and clinical experimental primate studies in craniofaciodental biology: a personal historical review. PMID- 10368099 TI - Malignant ameloblastoma: a case study and review. PMID- 10368100 TI - New orbital prosthesis with a blinking eyelid: report of a case. PMID- 10368101 TI - Mesenchymal chondrosarcoma of the maxilla: report of a case. PMID- 10368102 TI - Persistent sialadenitis after radioactive iodine therapy: report of two cases. PMID- 10368103 TI - Post-traumatic parotid sialocele: report of two cases. PMID- 10368104 TI - Nonfluent aphasia after closed head trauma: report of a case. PMID- 10368105 TI - Getting the picture correctly. PMID- 10368106 TI - Negative ultrasonic findings in patients with odontogenic infections. PMID- 10368108 TI - Linkage of a gene causing familial focal segmental glomerulosclerosis to chromosome 11 and further evidence of genetic heterogeneity. AB - Focal segmental glomerulosclerosis (FSGS) is a pathological entity characterized by proteinuria, nephrotic syndrome, and the progressive loss of renal function. It is a common cause of end-stage renal disease (ESRD). Recently, familial forms of FSGS have been identified. Two families with autosomal dominant FSGS were evaluated for linkage using 351 genomic microsatellite markers. Linkage, multipoint analysis, and tests for heterogeneity were performed on the subsequent results. In addition, three small families were used for haplotype analysis. Evidence for linkage was found on chromosome 11q21-q22 for the largest family, with a maximum lod score of 9.89. The gene is currently localized to an 18-cM area between flanking markers D11S2002 and D11S1986. The disease in a second family was not linked to this locus or to a previously described locus on chromosome 19q13. There were no shared haplotypes among affected individuals in the three smaller families. Our findings demonstrate that genetic heterogeneity is prevalent in FSGS in that at least three genes cause the FSGS phenotype. Identification of the genes that cause familial FSGS will provide valuable insights into the molecular basis and pathophysiology of FSGS. PMID- 10368110 TI - Thomas W. mattingly PMID- 10368109 TI - Circulation online only : june 15, 1999 PMID- 10368111 TI - Nitric oxide synthases in the failing human heart: a doubled-edged sword? PMID- 10368112 TI - Intravascular ultrasound evidence for coarctation causing symptomatic renal artery stenosis. AB - BACKGROUND: A recent study of human cadaveric renal arteries revealed that renal artery narrowing could be due not only to atherosclerotic plaque compensated for by adaptive remodeling, but also to hitherto undescribed focal narrowing of an otherwise normal renal arterial wall (ie, coarctation). The present study investigated whether vessel coarctation could be identified in patients with symptomatic renal artery stenosis (RAS). METHODS AND RESULTS: Consecutive symptomatic patients with angiographically proven atherosclerotic RAS who were referred for stent placement were studied by 30-MHz intravascular ultrasound before intervention (n=18) or after predilatation (n=18). Analysis included assessment of the media-bounded area and plaque area (PLA) at the most stenotic site and at a distal reference site (most distal cross-section in the main renal artery with normal appearance). Coarctation was considered present whenever the target/reference media-bounded area was 3-fold fewer monocytes than Ad.betaGal- or sham-treated arteries. CONCLUSIONS: NO synthase gene therapy rapidly ameliorates several markers of atherosclerosis in the cholesterol-fed rabbit. PMID- 10368114 TI - Effects of acute angiotensin II type 1 receptor antagonism and angiotensin converting enzyme inhibition on plasma fibrinolytic parameters in patients with heart failure. AB - BACKGROUND: Angiotensin converting enzyme (ACE) inhibition after myocardial infarction is associated with an improvement in plasma fibrinolytic parameters. The aim of the present study was to determine whether acute ACE inhibition and angiotensin II type 1 (AT1) receptor antagonism have similar effects in patients with heart failure. METHODS AND RESULTS: Twenty patients with moderately severe chronic heart failure received enalapril 10 mg and losartan 50 mg on 2 separate occasions in a single-blind, randomized, crossover design. Plasma tissue plasminogen activator (t-PA) and plasminogen activator inhibitor type 1 (PAI-1) antigen and activity were measured at baseline and 6 hours after the dose. Acute administration of losartan but not of enalapril reduced plasma t-PA (11%; P=0.003) and PAI-1 (38%; P<0.001) antigen concentrations, which was associated with increases in t-PA (29%; P=0.03) and decreases in PAI-1 (48%; P=0.01) activity. Changes in plasma fibrinolytic parameters were more marked during losartan treatment (P<0.02), with a 3-fold greater reduction in plasma PAI-1 antigen concentrations (P<0.05). CONCLUSIONS: Acute AT1 antagonism in patients with heart failure is associated with a significant improvement in plasma fibrinolytic parameters that is greater than during ACE inhibition. These beneficial effects of AT1 antagonism and ACE inhibition would therefore appear to be mediated principally through suppression of angiotensin II. PMID- 10368115 TI - Comparing AMI mortality among hospitals in patients 65 years of age and older: evaluating methods of risk adjustment. AB - BACKGROUND: Interest in the reporting of risk-adjusted outcomes for patients with acute myocardial infarction is growing. A useful risk-adjustment model must balance parsimony and ease of data collection with predictive ability. METHODS AND RESULTS: From our analysis of 82 359 patients >/=65 years of age admitted with acute myocardial infarction to 2401 hospitals, we derived a parsimonious model that predicts 30-day mortality. The model was validated on a similar group of 78 699 patients from 2386 hospitals. Of the 73 candidate predictor variables examined, 7 variables describing patient characteristics on arrival were selected for inclusion in the final model: age, cardiac arrest, anterior or lateral location of myocardial infarction, systolic blood pressure, white blood cell count, serum creatinine, and congestive heart failure. The area under the receiver-operating characteristic curve for the final model was 0.77 in the derivation cohort and 0.77 in the validation cohort. The rankings of hospitals by performance (in deciles) with this model were most similar to a comprehensive 27 variable model based on medical chart review and least similar to models based on administrative billing codes. CONCLUSIONS: A simple 7-variable risk model performs as well as more complex models in comparing hospital outcomes for acute myocardial infarction. Although there is a continuing need to improve methods of risk adjustment, our results provide a basis for hospitals to develop a simple approach to compare outcomes. PMID- 10368116 TI - Effect of pacing chamber and atrioventricular delay on acute systolic function of paced patients with congestive heart failure. The Pacing Therapies for Congestive Heart Failure Study Group. The Guidant Congestive Heart Failure Research Group. AB - BACKGROUND: Previous studies of pacing therapy for dilated congestive heart failure (CHF) have not established the relative importance of pacing site, AV delay, and patient heterogeneity on outcome. These variables were compared by a novel technique that evaluated immediate changes in hemodynamic function during brief periods of atrial-synchronous ventricular pacing. METHODS AND RESULTS: Twenty-seven CHF patients with severe left ventricular (LV) systolic dysfunction and LV conduction disorder were implanted with endocardial pacing leads in the right atrium and right ventricle (RV) and an epicardial lead on the LV and instrumented with micromanometer catheters in the LV, aorta, and RV. Patients in normal sinus rhythm were stimulated in the RV, LV, or both ventricles simultaneously (BV) at preselected AV delays in a repeating 5-paced/15-nonpaced beat sequence. Maximum LV pressure derivative (LV+dP/dt) and aortic pulse pressure (PP) changed immediately at pacing onset, increasing at a patient specific optimal AV delay in 20 patients with wide surface QRS (180+/-22 ms) and decreasing at short AV delays in 5 patients with narrower QRS (128+/-12 ms) (P<0.0001). Overall, BV and LV pacing increased LV+dP/dt and PP more than RV pacing (P<0.01), whereas LV pacing increased LV+dP/dt more than BV pacing (P<0.01). CONCLUSIONS: In this population, CHF patients with sufficiently wide surface QRS benefit from atrial-synchronous ventricular pacing, LV stimulation is required for maximum acute benefit, and the maximum benefit at any site occurs with a patient-specific AV delay. PMID- 10368117 TI - Blood flow dynamics in heart failure. AB - BACKGROUND: Exercise intolerance in heart failure (HF) may be due to inadequate vasodilation, augmented vasoconstriction, and/or altered muscle metabolic responses that lead to fatigue. METHODS AND RESULTS: Vascular and metabolic responses to rhythmic forearm exercise were tested in 9 HF patients and 9 control subjects (CTL) during 2 protocols designed to examine the effect of HF on the time course of oxygen delivery versus uptake (protocol 1) and on vasoconstriction during exercise with 50 mm Hg pressure about the forearm to evoke a metaboreflex (protocol 2). In protocol 1, venous lactate and H+ were greater at 4 minutes of exercise in HF versus CTL (P<0.05) despite similar blood flow and oxygen uptake responses. In protocol 2, mean arterial pressure increased similarly in each group during ischemic exercise. In CTL, forearm blood flow and vascular conductance were similar at the end of ischemic and ambient exercise. In HF, forearm blood flow and vascular conductance were reduced during ischemic exercise compared with the ambient trial. CONCLUSIONS: Intrinsic differences in skeletal muscle metabolism, not vasodilatory dynamics, must account for the augmented glycolytic metabolic responses to moderate-intensity exercise in class II and III HF. The inability to increase forearm vascular conductance during ischemic handgrip exercise, despite a normal pressor response, suggests that enhanced vasoconstriction of strenuously exercising skeletal muscle contributes to exertional fatigue in HF. PMID- 10368118 TI - Endomyocardial nitric oxide synthase and left ventricular preload reserve in dilated cardiomyopathy. AB - BACKGROUND: Patients with heart failure have modified myocardial expression of nitric oxide synthase (NOS), as is evident from induction of calcium-insensitive NOS isoforms. The functional significance of this modified NOS gene expression for left ventricular (LV) contractile performance was investigated in patients with dilated nonischemic cardiomyopathy. METHODS AND RESULTS: In patients with dilated, nonischemic cardiomyopathy, invasive measures of LV contractile performance were derived from LV microtip pressure recordings and angiograms and correlated with intensity of gene expression of inducible (NOS2) and constitutive (NOS3) NOS isoforms in simultaneously procured LV endomyocardial biopsies (n=20). LV endomyocardial expression of NOS2 was linearly correlated with LV stroke volume (P=0.001; r=0.66), LV ejection fraction (P=0.007; r=0.58), and LV stroke work (P=0.003; r=0.62). In patients with elevated LV end-diastolic pressure (>16 mm Hg), a closer correlation was observed between endomyocardial expression of NOS2 and LV stroke volume (P=0.001; r=0.74), LV ejection fraction (P=0.0007; r=0.77), and LV stroke work (r=0.82; P=0.0002). LV endomyocardial expression of NOS3 was linearly correlated with LV stroke volume (P=0.01; r=0.53) and LV stroke work (P=0.01; r=0.52). To establish the role of nitric oxide (NO) as a mediator of the observed correlations, substance P (which causes endothelial release of NO) was infused intracoronarily (n=12). In patients with elevated LV end diastolic pressure, an intracoronary infusion of substance P increased LV stroke volume from 72+/-13 to 91+/-16 mL (P=0.06) and LV stroke work from 67+/-11 to 90+/-15 g. m (P=0.03) and shifted the LV end-diastolic pressure-volume relation to the right. CONCLUSIONS: In patients with dilated cardiomyopathy, an increase in endomyocardial NOS2 or NOS3 gene expression augments LV stroke volume and LV stroke work because of a NO-mediated rightward shift of the diastolic LV pressure volume relation and a concomitant increase in LV preload reserve. PMID- 10368119 TI - Angiographic anatomy of the inferior right atrial isthmus in patients with and without history of common atrial flutter. AB - BACKGROUND: Although most ablative procedures undertaken for common atrial flutter target the inferior right atrial isthmus, comparative studies of the morphology of this area are lacking. Our study examines its angiographic anatomy, making correlations with postmortem specimens, to provide a better understanding of the anatomic substrate of this arrhythmia. METHODS AND RESULTS: The gross morphological features and dimensions of the area between the orifice of the inferior caval vein and the attachment of the septal leaflet of the tricuspid valve were determined from angiograms made in 23 patients with documented atrial flutter and 30 control subjects. For comparison, we studied 20 normal heart specimens. When viewed in right anterior oblique projection, 2 morphologically distinct areas were identified. In the specimens, the inferior isthmus measured a mean length of 30+/-4 mm, not significantly different from the dimensions obtained from angiograms of control subjects. The mean length of the isthmus, however, was greater in patients with common atrial flutter than those without (37+/-8 versus 28+/-6 mm). Patients with atrial flutter and structural heart disease had an even longer isthmus than those with flutter alone (39. 6+/-8 versus 33+/-7 mm). Compared with those without flutter, the atrial diameter was also larger in patients with flutter (57.6+/-9 versus 48.5+/-6 mm). Reevaluation carried out at follow-up 10+/-2 months after ablation did not show any reduction in atrial size, although contractility improved. CONCLUSIONS: The inferior isthmus and right atrium in patients with common atrial flutter were significantly larger than those in a control population. PMID- 10368120 TI - Short-term effect of atrial fibrillation on atrial contractile function in humans. AB - BACKGROUND: Conversion of chronic atrial fibrillation (AF) is associated with atrial stunning, but the short-term effect of a brief episode of AF on left atrial appendage (LAA) emptying velocity is unknown. The purpose of this study was to determine whether a short episode of AF affects left atrial function and whether verapamil modifies this effect. METHODS AND RESULTS: The subjects of this study were 19 patients without structural heart disease undergoing an electrophysiology procedure. In 13 patients, LAA emptying velocity was measured by transesophageal echocardiography in the setting of pharmacological autonomic blockade before, during, and after a short episode of AF. During sinus rhythm, the baseline LAA emptying velocity was measured 5 times and averaged. AF was then induced by rapid right atrial pacing. After either spontaneous or electrical conversion, LAA emptying velocity was measured immediately on resumption of sinus rhythm and every minute thereafter. The mean duration of AF was 15.3+/-3.8 minutes. The mean baseline emptying velocity was 70+/-20 cm/s. The first post-AF emptying velocity was 63+/-20 cm/s (P=0.02 versus baseline emptying velocity). The post-AF emptying velocity returned to the baseline emptying velocity value after 3.0 minutes. The mean percent reduction in post-AF emptying velocity was 9.7+/-21% (range, 15% increase to 56% decrease). A second group of 6 patients were pretreated with verapamil (0.1-mg/kg IV bolus followed by an infusion of 0.005 mg. kg-1. min-1). In these patients, the first post-AF emptying velocity, 58+/-14 cm/s, was not significantly different from the pre-AF emptying velocity, 60+/-13 cm/s (P=0.08). CONCLUSIONS: In humans, several minutes of AF may be sufficient to induce atrial contractile dysfunction after cardioversion. When atrial contractile dysfunction occurs, there is recovery of AF within several minutes. AF-induced contractile dysfunction is attenuated by verapamil and may be at least partially mediated by cellular calcium overload. PMID- 10368121 TI - Characterization of different subsets of atrial fibrillation in general practice in France: the ALFA study. The College of French Cardiologists. AB - BACKGROUND: The clinical presentation and causes of atrial fibrillation (AF) in the 1990s may differ from AF seen 2 to 3 decades ago. It was the objective of this prospective study to characterize various clinical presentations and underlying conditions of patients with AF observed in general practice in France. METHODS AND RESULTS: The study population comprised 756 patients (19 to 95 years of age) with electrocardiographically documented AF subdivided into paroxysmal (<7 days), chronic (last episode >1 month) and recent onset AF(persistent >7 days and<1 month). Symptoms were present in 670 patients (88.6%). The relative prevalences of paroxysmal, chronic, and recent onset AF were 22.1%, 51.4%, and 26.4%, respectively. Cardiac disorders, present in 534 patients (70.6%), included hypertension (39.4%), coronary artery disease (16.6%), and myocardial diseases (15.3%) as the most common. Rheumatic valvular disease represented a common cause in women (25. 0%) but not in men (8.0%). The paroxysmal group differed by a high percentage of palpitations (79.0%) and a low percentage of underlying heart disease (53.9%). With a mean follow-up of 8.6+/-3.7 months, 28 patients (3.7%) died, including 6 fatal cerebrovascular accidents. Among the 728 patients who survived, congestive heart failure occurred in 30 patients (4.1%), and embolic complications occurred in 13 patients (1.8%). In the paroxysmal AF group, 13 patients (8.0%) developed chronic AF and 51 (31.3%) had AF recurrences. At the time of follow-up, 53 patients (14.3%) from the chronic AF group and 108 patients (55.7%) from the recent onset AF group were in sinus rhythm. CONCLUSIONS: This large-scale study establishes the current demographic profile of out-of-hospital patients with AF and highlights some of the changes that have occurred in the past decades, including a particular shift in cardiac causes toward nonrheumatic AF. This study also demonstrates significant differences between various subsets of AF. PMID- 10368122 TI - Char syndrome, an inherited disorder with patent ductus arteriosus, maps to chromosome 6p12-p21. AB - BACKGROUND: Patent ductus arteriosus (PDA) is a relatively common form of congenital heart disease. Although polygenic inheritance has been implicated, no specific gene defects causing PDA have been identified to date. Thus, a positional cloning strategy was undertaken to determine the gene responsible for the Char syndrome, an autosomal dominant disorder characterized by PDA, facial dysmorphism, and hand anomalies. METHODS AND RESULTS: A genome scan was performed with 46 members of 2 unrelated families in which the disease was fully penetrant but the phenotype differed. Significant linkage was achieved with several polymorphic DNA markers mapping to chromosome 6p12-p21 (maximal 2-point LOD score of 8.39 with D6S1638 at theta=0.00). Haplotype analysis identified recombinant events that defined the Char syndrome locus with high probability to a 3. 1-cM region between D6S459/D6S1632/D6S1541 and D6S1024. CONCLUSIONS: A familial syndrome in which PDA is a common feature was mapped to a narrow region of chromosome 6p12-p21. Additional analysis with other families and polymorphic markers as well as evaluation of potential candidate genes should lead to the identification of the Char syndrome gene, which will provide insights into cardiogenesis as well as limb and craniofacial development. PMID- 10368123 TI - Tissue inhibition of angiotensin-converting enzyme activity stimulates angiogenesis in vivo. AB - BACKGROUND: Endothelial cells (ECs) represent the critical cellular element responsible for postnatal angiogenesis. Because ACE inhibitors may favorably affect endothelial function, we investigated the hypothesis that administration of the ACE inhibitor quinaprilat could enhance angiogenesis in vivo. METHODS AND RESULTS: Ten days after resection of 1 femoral artery, New Zealand White (NZW) rabbits were randomly assigned to receive recombinant human vascular endothelial growth factor (rhVEGF) administered as a single intra-arterial injection (n=6), quinaprilat (n=8) or captopril (n=7) administered as a daily subcutaneous injection, or no treatment (controls, n=6). Angiogenesis was monitored in vivo by measurement of blood pressure, vasoreactivity, and resistance in ischemic versus normal limbs at day 10 (D10) and D40; angiographic studies to identify sites of neovascularization were performed at D10 and D40, and morphometric analysis of capillary density in the ischemic limb was performed at necropsy (D40). Both functional and morphological outcomes documented augmented angiogenesis in quinaprilat-treated rabbits similar to that observed for rhVEGF and superior to that observed with either captopril or no drug (controls). Residual ACE activity was equivalent for the captopril and quinaprilat groups in plasma (42.54+/-0.03% versus 41.53+/-0.02%, P=NS) but not in tissue, where quinaprilat lowered ACE activity significantly (P<0.01) compared with captopril (13% versus 61%). CONCLUSIONS: ACE inhibition with quinaprilat promotes angiogenesis in a rabbit model of hindlimb ischemia. Thus, nonsulfhydryl ACE inhibitors with high tissue affinity may be potentially useful for therapeutic angiogenesis in ischemic tissues. Moreover, previous evidence that ACE inhibition benefits patients with myocardial ischemia may be due in part to augmented collateral development. PMID- 10368124 TI - Plasminogen activator inhibitor-1 is a major determinant of arterial thrombolysis resistance. AB - BACKGROUND: Platelet-rich thrombi are resistant to lysis by tissue plasminogen activator (tPA). Plasminogen activator inhibitor-1 (PAI-1), a rapid inhibitor of tPA, may contribute to arterial thrombolysis resistance. However, few data are available regarding the effect of PAI-1 on arterial thrombolysis in animals. We used a murine carotid injury model to test the hypothesis that PAI-1 inhibits thrombolysis mediated by pharmacological concentrations of tPA. METHODS AND RESULTS: Platelet-rich thrombi were induced in wild-type mice (PAI-1 +/+; n=11) and PAI-1-deficient mice (PAI-1 -/-; n=11) with ferric chloride. Baseline carotid blood flows and mean occlusion times did not differ between PAI-1 +/+ and PAI-1 /- mice. Clot lysis was induced by infusion of heparin (200 U/kg bolus, 70 U. kg 1. h-1 drip), human plasminogen (50 mg/kg), and tPA at 20 (n=10) or 100 (n=12) microg. kg-1. min-1. Mean plasma tPA antigens were 2.7 microg/mL (tPA infusion, 20 microg. kg-1. min-1) and 5.5 microg/mL (tPA infusion, 100 microg. kg-1. min 1), with no significant differences between PAI-1 +/+ mice and PAI-1 -/- mice. Reperfusion after tPA 20 microg. kg-1. min-1 occurred in 1 of 5 PAI-1 +/+ mice versus 5 of 5 PAI-1 -/- mice (P=0.0006). Reperfusion occurred in all mice that received tPA 100 microg. kg-1. min-1, but reperfusion times were significantly shorter in PAI-1 -/- mice (17. 8+/-2.6 minutes, n=6) than in PAI-1 +/+ mice (35.7+/-5.1 minute, n=6; P=0.01). Histological analyses confirmed that carotid thrombi were platelet rich and that PAI-1 was distributed uniformly throughout thrombi from PAI-1 +/+ mice. Lysates of PAI-1 +/+ platelets inhibited human tPA, whereas PAI-1 -/- platelet lysates did not. CONCLUSIONS: PAI-1 is a major determinant of the resistance of platelet-rich arterial thrombi to lysis by pharmacological concentrations of tPA. Strategies to inhibit or resist PAI-1 may enhance thrombolysis. PMID- 10368125 TI - Reduction in lipopolysaccharide-induced thrombocytopenia by triflavin in a rat model of septicemia. AB - BACKGROUND: Thrombocytopenia frequently occurs early in the course of Gram negative bacterial infections. Triflavin, an Arg-Gly-Asp-containing disintegrin, has been suggested to interfere with the interaction of fibrinogen with the glycoprotein IIb/IIIa complex. The present study was undertaken to determine whether triflavin could prevent thrombocytopenia in lipopolysaccharide (LPS) treated rats. METHODS AND RESULTS: In this study, 51Cr-labeled platelets were used to assess blood and tissue platelet accumulation after LPS challenge. The administration of LPS (4 mg/kg IV bolus) for 4 hours induced a reduction in radiolabeled platelets in blood and an obvious accumulation of platelets in liver. Triflavin (500 microg/kg) but not GRGDS (20 mg/kg) significantly prevented the alteration of radiolabeled platelet distribution in blood and liver when induced by LPS. Furthermore, triflavin but not GRGDS markedly suppressed the elevation in plasma thromboxane B2 concentration within the 4-hour period of LPS administration. In LPS-treated rats, the 5-hydroxytryptamine level was lower in the blood and higher in the liver compared with levels in normal saline-treated rats. Pretreatment with triflavin (500 microg/kg) significantly reversed the 5 hydroxytryptamine concentration in blood and liver of LPS-treated rats. In histological examinations and platelet adhesion assay, triflavin markedly inhibited the adhesion of platelets to subendothelial matrixes in vivo and in vitro. CONCLUSIONS: The results indicate that triflavin effectively prevents thrombocytopenia, possibly through the following 2 mechanisms: (1) Triflavin markedly inhibits platelet aggregation, resulting in decreased thromboxane A2 formation. (2) It inhibits the adhesion of platelets to subendothelial matrixes, thereby leading to a reversal in the distribution of platelets in blood and liver in LPS-treated rats. PMID- 10368126 TI - Matrix metalloproteinase inhibition attenuates early left ventricular enlargement after experimental myocardial infarction in mice. AB - BACKGROUND: Extracellular matrix synthesis and degradation contribute to the morphological changes that occur after myocardial infarction (MI). METHODS AND RESULTS: We tested the hypothesis that inhibition of matrix metalloproteinases (MMPs) attenuates left ventricular remodeling in experimental MI. Seventy-one male FVB mice that survived ligation of the left anterior coronary artery were randomized to a broad-spectrum MMP inhibitor (CP-471,474) or placebo by gavage. Echocardiographic studies were performed before randomization (within 24 hours of surgery) and 4 days later and included short-axis imaging at the midpapillary and apical levels. Infarction as defined by wall motion abnormality was achieved in 79% of the procedures (n=56), and mortality rate during the 4-day protocol was 23% (9 of 36 on treatment vs 7 of 35 on placebo; P=NS). Baseline end-diastolic and end-systolic dimensions and areas were similar (P=NS) between treated and placebo groups. At follow-up, infarcted mice allocated to MMP inhibitor had significantly smaller increases in end-systolic and end-diastolic dimensions and areas at both midpapillary and apical levels compared with infarcted mice allocated to placebo (all P<0.05). In addition, infarcted animals that received MMP inhibitor had no change in fractional shortening (-3+/-13%), whereas animals that received placebo had a decrease in fractional shortening (-12+/-12%) (P<0.05). In an analysis stratified by baseline end-diastolic area, the effects of MMP inhibition on the changes in end-systolic area and end-diastolic area were most prominent in animals that had more initial left ventricular dilatation (both P<0.05). CONCLUSIONS: -Administration of an MMP inhibitor attenuates early left ventricular dilation after experimental MI in mice. Further studies in genetically altered mice and other models will improve understanding of the role of MMPs in left ventricular remodeling. PMID- 10368127 TI - Increased cardiomyocyte apoptosis and changes in proapoptotic and antiapoptotic genes bax and bcl-2 during left ventricular adaptations to chronic pressure overload in the rat. AB - BACKGROUND: Left ventricular hypertrophy (LVH) represents both an adaptive response to increased cardiac work load and a precursor state of heart failure. Recent evidence linked cardiac myocyte death by apoptosis with LVH and heart failure. It remained unclear, however, whether apoptosis participated in the transition from LVH to left ventricular dysfunction (LVD). METHODS AND RESULTS: Cardiac myocyte apoptotic events and changes in apoptosis-specific genes were studied in a rat model of chronic pressure overload induced by transverse aortic constriction. The changes in left ventricular geometry and function were assessed by echocardiography. Transverse aortic constriction rats progressively developed "concentric" LVH and subsequently, LVD. A similar distribution of LVH and LVD was found 18 weeks after surgery. At this time point, we determined the occurrence of myocyte apoptosis by DNA laddering, in situ DNA TUNEL labeling, and light and electron microscopy. The monitoring of proapoptotic and antiapoptotic genes was determined by Western blot and immunohistochemistry. Our data demonstrated that cardiomyocyte apoptotic events increased from virtually undetectable (in sham operated controls, SH) to 0.8/10(3) and 1.5/10(3) positive nuclei in LVH and LVD, respectively. Fibrosis also increased in the subendocardial and midwall regions of LVH and LVD rats compared with SH. Expression of the proapoptotic gene bax increased, whereas that of antiapoptotic gene bcl-2 decreased in LVH and LVD compared with SH. CONCLUSIONS: These data suggest that in response to chronic pressure overload, cardiomyocyte-specific apoptosis contributed to the transition from LVH to LVD. LVH and LVD were accompanied by a dramatic cardiomyocyte upregulation of the proapoptotic gene bax and reduced bcl-2/bax ratio, predisposing cardiomyocytes to apoptosis. PMID- 10368128 TI - Opening of potassium channels: the common cardioprotective link between preconditioning and natural hibernation? AB - BACKGROUND: The tolerance of hibernating mammals to cold hypoxia is related to a factor similar to agonists of delta-opioid receptors. This study was designed to assess whether activation of these receptors could reproduce the protection conferred by ischemic preconditioning and whether such cardioprotection was similarly mediated by an opening of ATP-sensitive potassium (KATP) channels. METHODS AND RESULTS: Thirty-two isolated rat hearts were arrested with and stored in Celsior at 4 degrees C for 5 hours before being reperfused for 2 hours. They were divided into 4 equal groups. Group 1 hearts served as controls. In group 2, ischemic preconditioning was elicited by two 5-minute global ischemia periods interspersed with 5 minutes of reperfusion before arrest. In group 3, hearts were pharmacologically preconditioned with a 15-minute infusion of the delta-opioid receptor agonist D-Ala2-D-Leu5-enkephalin (DADLE; 200 micromol/L). In group 4, the protocol was similar to group 3 except that infusion of DADLE was preceded by infusion of the KATP blocker glibenclamide (50 micromol/L). The salutary effects of both forms of preconditioning were primarily manifest as a better preservation of diastolic function, a reduced myocardial edema, and reduced creatine kinase leakage. This protection was abolished by administration of glibenclamide before DADLE. CONCLUSIONS: These data suggest that activation of delta-opioid receptors improves recovery of cold-stored hearts to a similar extent as ischemic preconditioning, most likely through an opening of KATP channels. This provides a rationale for improving the preservation of hearts for transplantation by pharmacologically duplicating the common pathway to natural hibernation and preconditioning. PMID- 10368129 TI - Three-dimensional visualization of recurrent coarctation of the aorta by electron beam tomography and MRI. PMID- 10368131 TI - Myocardial perfusion in acute coronary syndrome. PMID- 10368133 TI - A high incidence of prophage carriage among natural isolates of Streptococcus pneumoniae. AB - The majority (591 of 791, or 76%) of Streptococcus pneumoniae clinical isolates examined showed the presence of two or more chromosomal SmaI fragments that hybridized with the lytA-specific DNA probe. Only one of these fragments, frequently having an approximate molecular size of 90 kb, was shown to carry the genetic determinant of the pneumococcal autolysin (N-acetylmuramic acid-L-alanine amidase). Strains carrying multiple copies of lytA homologues included both antibiotic-susceptible and -resistant isolates as well as a number of different serotypes and strains recovered from geographic sites on three continents. Mitomycin C treatment of strains carrying several lytA-hybridizing fragments caused the appearance of extrachromosomal DNA hybridizing to the lytA gene, followed by lysis of the bacteria. Such lysates contained phage particles detectable by electron microscopy. The findings suggest that the lytA-hybridizing fragments in excess of the host lytA represent components of pneumococcal bacteriophages. The high proportion of clinical isolates carrying multiple copies of lytA indicates the widespread occurrence of lysogeny, which may contribute to genetic variation in natural populations of pneumococci. PMID- 10368132 TI - Archaebacteria then ... Archaes now (are there really no archaeal pathogens?). PMID- 10368134 TI - The mdoC gene of Escherichia coli encodes a membrane protein that is required for succinylation of osmoregulated periplasmic glucans. AB - Osmoregulated periplasmic glucans (OPGs) of Escherichia coli are anionic oligosaccharides that accumulate in the periplasmic space in response to low osmolarity of the medium. Their anionic character is provided by the substitution of the glucosidic backbone by phosphoglycerol originating from the membrane phospholipids and by succinyl residues from unknown origin. A phosphoglycerol transferase-deficient mdoB mutant was subjected to Tn5 transposon mutagenesis, and putative mutant clones were screened for changes in the anionic character of OPGs by thin-layer chromatography. One mutant deficient in succinylation of OPGs was obtained, and the gene inactivated in this mutant was characterized and named mdoC. mdoC, which encodes a membrane-bound protein, is closely linked to the mdoGH operon necessary for the synthesis of the OPG backbone. PMID- 10368135 TI - A Bacillus subtilis secreted protein with a role in endospore coat assembly and function. AB - Bacterial endospores are encased in a complex protein coat, which confers protection against noxious chemicals and influences the germination response. In Bacillus subtilis, over 20 polypeptides are organized into an amorphous undercoat, a lamellar lightly staining inner structure, and an electron-dense outer coat. Here we report on the identification of a polypeptide of about 30 kDa required for proper coat assembly, which was extracted from spores of a gerE mutant. The N-terminal sequence of this polypeptide matched the deduced product of the tasA gene, after removal of a putative 27-residue signal peptide, and TasA was immunologically detected in material extracted from purified spores. Remarkably, deletion of tasA results in the production of asymmetric spores that accumulate misassembled material in one pole and have a greatly expanded undercoat and an altered outer coat structure. Moreover, we found that tasA and gerE mutations act synergistically to decrease the efficiency of spore germination. We show that tasA is the most distal member of a three-gene operon, which also encodes the type I signal peptidase SipW. Expression of the tasA operon is enhanced 2 h after the onset of sporulation, under the control of sigmaH. When tasA transcription is uncoupled from sipW expression, a presumptive TasA precursor accumulates, suggesting that its maturation depends on SipW. Mature TasA is found in supernatants of sporulating cultures and intracellularly from 2 h of sporulation onward. We suggest that, at an early stage of sporulation, TasA is secreted to the septal compartment. Later, after engulfment of the prespore by the mother cell, TasA acts from the septal-proximal pole of the spore membranes to nucleate the organization of the undercoat region. TasA is the first example of a polypeptide involved in coat assembly whose production is not mother cell specific but rather precedes its formation. Our results implicate secretion as a mechanism to target individual proteins to specific cellular locations during the assembly of the bacterial endospore coat. PMID- 10368136 TI - Characterization of the vanD glycopeptide resistance gene cluster from Enterococcus faecium BM4339. AB - VanD-type resistance to glycopeptides in Enterococcus faecium BM4339 is due to constitutive synthesis of D-alanyl-D-lactate-terminating peptidoglycan precursors (B. Perichon, P. Reynolds, and P. Courvalin, Antimicrob. Agents Chemother. 41:2016-2018, 1997). The sequence of a 5,780-bp fragment was determined and revealed six open reading frames. The 3' distal part encoded the VanHD dehydrogenase, the VanD ligase, and the VanXD DD-dipeptidase, which were highly similar to the corresponding proteins in VanA and VanB types of resistance. The deduced VanYD protein was homologous to penicillin-binding proteins that display DD-carboxypeptidase activity. The 5' end coded for the putative VanRD-VanSD two component regulatory system. Due to a frameshift mutation in the chromosomal ddl gene, BM4339 produced an impaired D-alanine:D-alanine ligase. However, since expression of the resistance genes is constitutive, growth of E. faecium BM4339 was not dependent on the presence of glycopeptides in the culture medium. PMID- 10368137 TI - A response regulator that represses transcription of several virulence operons in the group A streptococcus. AB - A search for homologs of the Bacillus subtilis PhoP response regulator in the group A streptococcus (GAS) genome revealed three good candidates. Inactivation of one of these, recently identified as csrR (J. C. Levin and M. R. Wessels, Mol. Microbiol. 30:209-219, 1998), caused the strain to produce mucoid colonies and to increase transcription of hasA, the first gene in the operon for capsule synthesis. We report here that a nonpolar insertion in this gene also increased transcription of ska (encoding streptokinase), sagA (streptolysin S), and speMF (mitogenic factor) but did not affect transcription of slo (streptolysin O), mga (multiple gene regulator of GAS), emm (M protein), scpA (complement C5a peptidase), or speB or speC (pyrogenic exotoxins B and C). The amounts of streptokinase, streptolysin S, and capsule paralleled the levels of transcription of their genes in all cases. Because CsrR represses genes unrelated to those for capsule synthesis, and because CsrA-CsrB is a global regulatory system in Escherichia coli whose mechanism is unrelated to that of these genes in GAS, the locus has been renamed covR, for "control of virulence genes" in GAS. Transcription of the covR operon was also increased in the nonpolar insertion mutant, indicating that CovR represses its own synthesis as well. All phenotypes of the covR nonpolar insertion mutant were complemented by the covR gene on a plasmid. CovR acts on operons expressed both in exponential and in stationary phase, demonstrating that the CovR-CovS pathway is separate from growth phase dependent regulation in GAS. Therefore, CovR is the first multiple-gene repressor of virulence factors described for this important human pathogen. PMID- 10368138 TI - Nitrate and nitrite control of respiratory nitrate reduction in denitrifying Pseudomonas stutzeri by a two-component regulatory system homologous to NarXL of Escherichia coli. AB - Bacterial denitrification is expressed in response to the concurrent exogenous signals of low-oxygen tension and nitrate or one of its reduction products. The mechanism by which nitrate-dependent gene activation is effected was investigated in the denitrifying bacterium Pseudomonas stutzeri ATCC 14405. We have identified and isolated from this organism the chromosomal region encoding the two-component sensor-regulator pair NarXL and found that it is linked with the narG operon for respiratory nitrate reductase. The same region encodes two putative nitrate or nitrite translocases, NarK and NarC (the latter shows the highest similarity to yeast [Pichia] and plant [Nicotiana] nitrate transporters), and the nitrate regulated transcription factor, DnrE, of the FNR family. The roles of NarX and NarL in nitrate respiration were studied with deletion mutants. NarL activated the transcription of narG, narK, and dnrE but did not affect the denitrification regulons for the respiratory substrates nitrite, nitric oxide, and nitrous oxide. The promoters of narG, narK, and dnrE carry sequence motifs, TACYYMT, which correspond to the NarL recognition sequence established for Escherichia coli. The cellular response toward nitrate and nitrite was mediated by the sensor protein NarX, which discriminated weakly between these oxyanions. Our data show that the NarXL two-component regulatory system has been incorporated into the bacterial denitrification process of P. stutzeri for selective regulation of nitrate respiration. PMID- 10368139 TI - Role in cell permeability of an essential two-component system in Staphylococcus aureus. AB - A temperature-sensitive lethal mutant of Staphylococcus aureus was found to harbor a mutation in the uncharacterized two-component histidine kinase (HK) response regulator (RR) pair encoded by yycFG; orthologues of yycFG could be identified in the genomes of Bacillus subtilis and other gram-positive bacteria. Sequence analysis of the mutant revealed a point mutation resulting in a nonconservative change (Glu to Lys) in the regulator domain of the RR at position 63. To confirm that this signal transduction system was essential, a disrupted copy of either the RR (yycF) or the HK (yycG) was constructed with a set of suicide vectors and used to generate tandem duplications in the chromosome. Resolution of the duplications, leaving an insertion in either the yycF or the yycG coding region, was achieved only in the presence of an additional wild-type copy of the two open reading frames. Phenotypic characterization of the conditional lethal mutant showed that at permissive growth conditions, the mutant was hypersusceptible to macrolide and lincosamide antibiotics, even in the presence of the ermB resistance determinant. Other mutant phenotypes, including hypersensitivity to unsaturated long-chain fatty acids and suppression of the conditional lethal phenotype by high sucrose and NaCl concentrations, suggest that the role of the two-component system includes the proper regulation of bacterial cell wall or membrane composition. The effects of this point mutation are strongly bactericidal at the nonpermissive temperature, indicating that this pathway provides an excellent target for the identification of novel antibiotics. PMID- 10368140 TI - The ATP-dependent HslVU/ClpQY protease participates in turnover of cell division inhibitor SulA in Escherichia coli. AB - Escherichia coli mutants lacking activities of all known cytosolic ATP-dependent proteases (Lon, ClpAP, ClpXP, and HslVU), due to double deletions [DeltahslVU and Delta(clpPX-lon)], cannot grow at low (30 degrees C) or very high (45 degrees C) temperatures, unlike those carrying either of the deletions. Such growth defects were particularly marked when the deletions were introduced into strain MG1655 or W3110. To examine the functions of HslVU and other proteases further, revertants that can grow at 30 degrees C were isolated from the multiple-protease mutant and characterized. The revertants were found to carry a suppressor affecting either ftsZ (encoding a key cell division protein) or sulA (encoding the SulA inhibitor, which binds and inhibits FtsZ). Whereas the ftsZ mutations were identical to a mutation known to produce a protein refractory to SulA inhibition, the sulA mutations affected the promoter-operator region, reducing synthesis of SulA. These results suggested that the growth defect of the parental double-deletion mutant at a low temperature was due to the accumulation of excess SulA without DNA-damaging treatment. Consistent with these results, SulA in the double deletion mutant was much more stable than that in the Delta(clpPX-lon) mutant, suggesting that SulA can be degraded by HslVU. As expected, purified HslVU protease degraded SulA (fused to the maltose-binding protein) efficiently in an ATP-dependent manner. These results suggest that HslVU as well as Lon participates in the in vivo turnover of SulA and that HslVU becomes essential for growth when the Lon (and Clp) protease level is reduced below a critical threshold. PMID- 10368141 TI - Redundant in vivo proteolytic activities of Escherichia coli Lon and the ClpYQ (HslUV) protease. AB - The ClpYQ (HslUV) ATP-dependent protease of Escherichia coli consists of an ATPase subunit closely related to the Clp ATPases and a protease component related to those found in the eukaryotic proteasome. We found that this protease has a substrate specificity overlapping that of the Lon protease, another ATP dependent protease in which a single subunit contains both the proteolytic active site and the ATPase. Lon is responsible for the degradation of the cell division inhibitor SulA; lon mutants are UV sensitive, due to the stabilization of SulA. lon mutants are also mucoid, due to the stabilization of another Lon substrate, the positive regulator of capsule transcription, RcsA. The overproduction of ClpYQ suppresses both of these phenotypes, and the suppression of UV sensitivity is accompanied by a restoration of the rapid degradation of SulA. Inactivation of the chromosomal copy of clpY or clpQ leads to further stabilization of SulA in a lon mutant but not in lon+ cells. While either lon, lon clpY, or lon clpQ mutants are UV sensitive at low temperatures, at elevated temperatures the lon mutant loses its UV sensitivity, while the double mutants do not. Therefore, the degradation of SulA by ClpYQ at elevated temperatures is sufficient to lead to UV resistance. Thus, a protease with a structure and an active site different from those of Lon is capable of recognizing and degrading two different Lon substrates and appears to act as a backup for Lon under certain conditions. PMID- 10368142 TI - Structural and functional roles of the surface-exposed loops of the beta-barrel membrane protein OmpA from Escherichia coli. AB - The N-terminal domain of the OmpA protein from Escherichia coli, consisting of 170 amino acid residues, is embedded in the outer membrane, in the form of an antiparallel beta-barrel whose eight transmembrane beta-strands are connected by three short periplasmic turns and four relatively large surface-exposed hydrophilic loops. This protein domain serves as a paradigm for the study of membrane assembly of integral beta-structured membrane proteins. In order to dissect the structural and functional roles of the surface-exposed loops, they were shortened separately and in all possible combinations. All 16 loop deletion mutants assembled into the outer membrane with high efficiency and adopted the wild-type membrane topology. This systematic approach proves the absence of topogenic signals (e.g., in the form of loop sizes or charge distributions) in these loops. The shortening of surface-exposed loops did not reduce the thermal stability of the protein. However, none of the mutant proteins, with the exception of the variant with the fourth loop shortened, served as a receptor for the OmpA-specific bacteriophage K3. Furthermore, all loops were necessary for the OmpA protein to function in the stabilization of mating aggregates during F conjugation. An OmpA deletion variant with all four loops shortened, consisting of only 135 amino acid residues, constitutes the smallest beta-structured integral membrane protein known to date. These results represent a further step toward the development of artificial outer membrane proteins. PMID- 10368143 TI - The glucuronic acid utilization gene cluster from Bacillus stearothermophilus T 6. AB - A lambda-EMBL3 genomic library of Bacillus stearothermophilus T-6 was screened for hemicellulolytic activities, and five independent clones exhibiting beta xylosidase activity were isolated. The clones overlap each other and together represent a 23.5-kb chromosomal segment. The segment contains a cluster of xylan utilization genes, which are organized in at least three transcriptional units. These include the gene for the extracellular xylanase, xylanase T-6; part of an operon coding for an intracellular xylanase and a beta-xylosidase; and a putative 15.5-kb-long transcriptional unit, consisting of 12 genes involved in the utilization of alpha-D-glucuronic acid (GlcUA). The first four genes in the potential GlcUA operon (orf1, -2, -3, and -4) code for a putative sugar transport system with characteristic components of the binding-protein-dependent transport systems. The most likely natural substrate for this transport system is aldotetraouronic acid [2-O-alpha-(4-O-methyl-alpha-D-glucuronosyl)-xylotriose] (MeGlcUAXyl3). The following two genes code for an intracellular alpha glucuronidase (aguA) and a beta-xylosidase (xynB). Five more genes (kdgK, kdgA, uxaC, uxuA, and uxuB) encode proteins that are homologous to enzymes involved in galacturonate and glucuronate catabolism. The gene cluster also includes a potential regulatory gene, uxuR, the product of which resembles repressors of the GntR family. The apparent transcriptional start point of the cluster was determined by primer extension analysis and is located 349 bp from the initial ATG codon. The potential operator site is a perfect 12-bp inverted repeat located downstream from the promoter between nucleotides +170 and +181. Gel retardation assays indicated that UxuR binds specifically to this sequence and that this binding is efficiently prevented in vitro by MeGlcUAXyl3, the most likely molecular inducer. PMID- 10368144 TI - Modes of action of five different endopectate lyases from Erwinia chrysanthemi 3937. AB - Five endopectate lyases from the phytopathogenic bacterium Erwinia chrysanthemi, PelA, PelB, PelD, PelI, and PelL, were analyzed with respect to their modes of action on polymeric and oligomeric substrates (degree of polymerization, 2 to 8). On polygalacturonate, PelB showed higher reaction rates than PelD, PelI, and PelA, whereas the reaction rates for PelL were extremely low. The product progression during polygalacturonate cleavage showed a typical depolymerization profile for each enzyme and demonstrated their endolytic character. PelA, PelI, and PelL released oligogalacturonates of different sizes, whereas PelD and PelB released mostly unsaturated dimer and unsaturated trimer, respectively. Upon prolonged incubation, all enzymes degraded the primary products further, to unsaturated dimer and trimer, except for PelL, which degraded the primary products to unsaturated tetramer and pentamer in addition to unsaturated dimer and trimer. The bond cleavage frequencies on oligogalacturonates revealed differences in the modes of action of these enzymes that were commensurate with the product progression profiles. The preferential products formed from the oligogalacturonates were unsaturated dimer for PelD, unsaturated trimer for PelB, and unsaturated tetramer for PelI and PelL. For PelA, preferential products were dependent on the sizes of the oligogalacturonates. Whereas PelB and PelD displayed their highest activities on hexagalacturonate and tetragalacturonate, respectively, PelA, PelI, and PelL were most active on the octamer, the largest substrate used. The bond cleavage frequencies and reaction rates were used to estimate the number of subsites of each enzyme. PMID- 10368146 TI - Functioning of DcuC as the C4-dicarboxylate carrier during glucose fermentation by Escherichia coli. AB - The dcuC gene of Escherichia coli encodes an alternative C4-dicarboxylate carrier (DcuC) with low transport activity. The expression of dcuC was investigated. dcuC was expressed only under anaerobic conditions; nitrate and fumarate caused slight repression and stimulation of expression, respectively. Anaerobic induction depended mainly on the transcriptional regulator FNR. Fumarate stimulation was independent of the fumarate response regulator DcuR. The expression of dcuC was not significantly inhibited by glucose, assigning a role to DcuC during glucose fermentation. The inactivation of dcuC increased fumarate-succinate exchange and fumarate uptake by DcuA and DcuB, suggesting a preferential function of DcuC in succinate efflux during glucose fermentation. Upon overexpression in a dcuC promoter mutant (dcuC*), DcuC was able to compensate for DcuA and DcuB in fumarate-succinate exchange and fumarate uptake. PMID- 10368145 TI - The morphological transition of Helicobacter pylori cells from spiral to coccoid is preceded by a substantial modification of the cell wall. AB - The peptidoglycan (murein) of Helicobacter pylori has been investigated by high performance liquid chromatography and mass spectrometric techniques. Murein from H. pylori corresponded to the A1gamma chemotype, but the muropeptide elution patterns were substantially different from the one for Escherichia coli in that the former produced high proportions of muropeptides with a pentapeptide side chain (about 60 mol%), with Gly residues as the C-terminal amino acid (5 to 10 mol%), and with (1-->6)anhydro-N-acetylmuramic acid (13 to 18 mol%). H. pylori murein also lacks murein-bound lipoprotein, trimeric muropeptides, and (L-D) cross-linked muropeptides. Cessation of growth and transition to coccoid shape triggered an increase in N-acetylglucosaminyl-N-acetylmuramyl-L-Ala-D-Glu (approximately 20 mol%), apparently at the expense of monomeric muropeptides with tri- and tetrapeptide side chains. Muropeptides with (1-->6)anhydro-muramic acid and with Gly were also more abundant in resting cells. PMID- 10368147 TI - Proliferation of intrahyphal hyphae caused by disruption of csmA, which encodes a class V chitin synthase with a myosin motor-like domain in Aspergillus nidulans. AB - We have found that the Aspergillus nidulans csmA gene encodes a novel protein which consists of an N-terminal myosin motor-like domain and a C-terminal chitin synthase domain (M. Fujiwara, H. Horiuchi, A. Ohta, and M. Takagi, Biochem. Biophys. Res. Commun. 236:75-78, 1997). To clarify the roles of csmA in fungal morphogenesis, we constructed csmA null mutants. The growth rate of the mutant colonies was almost the same as that of the wild-type strain, but hyphal growth was severely inhibited when a chitin-binding reagent, Calcofluor white or Congo red, was added to the medium. Moreover, morphological abnormalities in tip growth and septum formation were identified microscopically. Proliferation of intracellular new hyphae, called intrahyphal hyphae, which behaved as intrinsic hyphae, was the most striking phenotypic feature among them. These phenotypes were not suppressed when the only chitin synthase domain of csmA was expressed under the control of the alcA promoter, whereas they were suppressed when the intact form of csmA was expressed. Therefore, it was concluded that the product of csmA (CsmA) has important roles in polarized cell wall synthesis and maintenance of cell wall integrity and that the myosin motor-like domain is indispensable for these functions. PMID- 10368149 TI - Insertional inactivation of Treponema denticola tap1 results in a nonmotile mutant with elongated flagellar hooks. AB - The treponemal fla operon is comprised of numerous motility-related genes; however, the initial gene of this operon, tap1, has no known function. A recently developed system to generate specific mutants in Treponema denticola was utilized to determine if Tap1 was essential for motility. T. denticola tap1 and flanking DNA were identified, cloned, and sequenced, and a suicide plasmid that contained tap1 interrupted with an erythromycin resistance cassette (ermF and ermAM) was constructed. Because of potential polar effects from this cassette, a second plasmid that contained tap1 interrupted with a modified erythromycin resistance cassette that lacked the putative ermF transcription terminator was constructed. Electroporation-mediated allelic exchange incorporated the interrupted tap1 genes into the T. denticola chromosome, creating Tap1-deficient mutants. Reverse transcriptase PCR revealed that the erythromycin resistance cassette within tap1 did not terminate fla operon transcription in either mutant. Moreover, the phenotypes of the two mutants were indistinguishable. These mutants lacked motion in liquid culture, were unable to spread on agar plates, and lacked flagellar filaments as determined by electron microscopy. Immunoblots revealed a marked reduction in detectable FlaB flagellar filament protein compared to that of wild type; however, flaB RNA was easily detectable, and transcription levels did not appear to be altered. The basis for the lack of filament protein expression is unknown. Immunoblotting also showed that the flagellar hook protein (FlgE) was synthesized in the Tap1-deficient mutant; however, electron microscopy revealed that the mutant possessed unusual elongated hooks of variable lengths. We propose that treponemal Tap1 is analogous to FliK, which is involved in monitoring the flagellar hook length of Salmonella typhimurium. PMID- 10368148 TI - Bacterioferritin A modulates catalase A (KatA) activity and resistance to hydrogen peroxide in Pseudomonas aeruginosa. AB - We have cloned a 3.6-kb genomic DNA fragment from Pseudomonas aeruginosa harboring the rpoA, rplQ, katA, and bfrA genes. These loci are predicted to encode, respectively, (i) the alpha subunit of RNA polymerase; (ii) the L17 ribosomal protein; (iii) the major catalase, KatA; and (iv) one of two iron storage proteins called bacterioferritin A (BfrA; cytochrome b1 or b557). Our goal was to determine the contributions of KatA and BfrA to the resistance of P. aeruginosa to hydrogen peroxide (H2O2). When provided on a multicopy plasmid, the P. aeruginosa katA gene complemented a catalase-deficient strain of Escherichia coli. The katA gene was found to contain two translational start codons encoding a heteromultimer of approximately 160 to 170 kDa and having an apparent Km for H2O2 of 44.7 mM. Isogenic katA and bfrA mutants were hypersusceptible to H2O2, while a katA bfrA double mutant demonstrated the greatest sensitivity. The katA and katA bfrA mutants possessed no detectable catalase activity. Interestingly, a bfrA mutant expressed only approximately 47% the KatA activity of wild-type organisms, despite possessing wild-type katA transcription and translation. Plasmids harboring bfrA genes encoding BfrA altered at critical amino acids essential for ferroxidase activity could not restore wild-type catalase activity in the bfrA mutant. RNase protection assays revealed that katA and bfrA are on different transcripts, the levels of which are increased by both iron and H2O2. Mass spectrometry analysis of whole cells revealed no significant difference in total cellular iron levels in the bfrA, katA, and katA bfrA mutants relative to wild-type bacteria. Our results suggest that P. aeruginosa BfrA may be required as one source of iron for the heme prosthetic group of KatA and thus for protection against H2O2. PMID- 10368150 TI - Importance of cis determinants and nitrogenase activity in regulated stability of the Klebsiella pneumoniae nitrogenase structural gene mRNA. AB - The Klebsiella pneumoniae nitrogen fixation (nif) mRNAs are unusually stable, with half-lives of 20 to 30 min under conditions favorable to nitrogen fixation (limiting nitrogen, anaerobiosis, temperatures of 30 degrees C). Addition of O2 or fixed nitrogen or temperature increases to 37 degrees C or more result in the dramatic destabilization of the nif mRNAs, decreasing the half-lives by a factor of 3 to 5. A plasmid expression system, independent of nif transcriptional regulation, was used to define cis determinants required for the regulated stability of the 5.2-kb nifHDKTY mRNA and to test the model suggested by earlier work that NifA is required in trans to stabilize nif mRNA under nif-derepressing conditions. O2 regulation of nifHDKTY mRNA stability is impaired in a plasmid containing a deletion of a 499-bp region of nifH, indicating that a site(s) required for the O2-regulated stability of the mRNA is located within this region. The simple model suggested from earlier work that NifA is required for stabilizing nif mRNA under conditions favorable for nitrogen fixation was disproved, and in its place, a more complicated model involving the sensing of nitrogenase activity as a component of the system regulating mRNA stability is proposed. Analysis of nifY mutants and overexpression suggests a possible involvement of the protein in this sensing process. PMID- 10368151 TI - A mutant Escherichia coli primase defective in elongation of primer RNA chains. AB - Earlier we showed by affinity cross-linking of initiating substrates to Escherichia coli primase that one or more of the residues Lys211, Lys229, and Lys241 were involved in the catalytic center of the enzyme (A. A. Mustaev and G. N. Godson, J. Biol. Chem. 270:15711-15718, 1995). We now demonstrate by mutagenesis that only Lys241 but not Lys211 and Lys229 is part of the catalytic center. Primase with a mutation of Arg to Lys at position 241 (defined as K241R primase) is almost unable to synthesize primer RNA (pRNA) on the single-stranded DNA-binding protein (SSB)/R199G4oric template. However, it is able to synthesize a pppApG dimer plus trace amounts of 8- to 11-nucleotide (nt) pRNA transcribed from the 5' CTG 3' pRNA initiation site on phage G4 oric DNA. The amount of dimer synthesized by K241R-primase is similar to that synthesized by the wild-type primase, demonstrating that the K241R mutant can initiate pRNA synthesis normally but is deficient in chain elongation. In the general priming system, the K241R primase also can synthesize only the dimer and very small amounts of 11-nt pRNA. The results of gel retardation experiments suggested that this deficiency in pRNA chain elongation of the K241R mutant primase is unlikely to be caused by impairment of the DNA binding activity. The K241R mutant primase, however, can still prime DNA synthesis in vivo and in vitro. PMID- 10368152 TI - Transcriptional organization and in vivo role of the Escherichia coli rsd gene, encoding the regulator of RNA polymerase sigma D. AB - The regulator of sigma D (Rsd) was identified as an RNA polymerase sigma70 associated protein in stationary-phase Escherichia coli with the inhibitory activity of sigma70-dependent transcription in vitro (M. Jishage and A. Ishihama, Proc. Natl. Acad. Sci. USA 95:4953-4958, 1998). Primer extension analysis of rsd mRNA indicated the presence of two promoters, sigmaS-dependent P1 and sigma70 dependent P2 with the gearbox sequence. To get insight into the in vivo role of Rsd, the expression of a reporter gene fused to either the sigma70- or sigmaS dependent promoter was analyzed in the absence of Rsd or the presence of overexpressed Rsd. In the rsd null mutant, the sigma70- and sigmaS-dependent gene expression was increased or decreased, respectively. On the other hand, the sigma70- or sigmaS-dependent transcription was reduced or enhanced, respectively, after overexpression of Rsd. The repression of the sigmaS-dependent transcription in the rsd mutant is overcome by increased production of the sigmaS subunit. Together these observations support the prediction that Rsd is involved in replacement of the RNA polymerase sigma subunit from sigma70 to sigmaS during the transition from exponential growth to the stationary phase. PMID- 10368153 TI - Transcriptional regulation in the hyperthermophilic archaeon Pyrococcus furiosus: coordinated expression of divergently oriented genes in response to beta-linked glucose polymers. AB - The genetic organization, expression, and regulation of the celB locus of the hyperthermophilic archaeon Pyrococcus furiosus were analyzed. This locus includes the celB gene, which codes for an intracellular beta-glucosidase, and a divergently orientated gene cluster, adhA-adhB-lamA, which codes for two alcohol dehydrogenases and an extracellular beta-1,3-endoglucanase that is transcribed as a polycistronic messenger (the lamA operon). During growth of P. furiosus on either the beta-1,4-linked glucose dimer cellobiose or the beta-1,3-linked glucose polymer laminarin, the activities of both beta-glucosidase and endoglucanase were increased at least fivefold compared with levels during growth on maltose or pyruvate. Northern blot analysis revealed an enhanced transcription of both the celB gene and the lamA operon in the presence of these glucose containing substrates. The in vivo and in vitro transcription initiation sites of both the celB gene and the lamA operon were identified 25 nucleotides downstream of conserved TATA box motifs. A number of repeating sequences have been recognized in the celB-adhA intergenic region, some of which might be part of a transcriptional regulator-binding site. PMID- 10368154 TI - Porphyrin-mediated binding to hemoglobin by the HA2 domain of cysteine proteinases (gingipains) and hemagglutinins from the periodontal pathogen Porphyromonas gingivalis. AB - Heme binding and uptake are considered fundamental to the growth and virulence of the gram-negative periodontal pathogen Porphyromonas gingivalis. We therefore examined the potential role of the dominant P. gingivalis cysteine proteinases (gingipains) in the acquisition of heme from the environment. A recombinant hemoglobin-binding domain that is conserved between two predominant gingipains (domain HA2) demonstrated tight binding to hemin (Kd = 16 nM), and binding was inhibited by iron-free protoporphyrin IX (Ki = 2.5 microM). Hemoglobin binding to the gingipains and the recombinant HA2 (rHA2) domain (Kd = 2.1 nM) was also inhibited by protoporphyrin IX (Ki = 10 microM), demonstrating an essential interaction between the HA2 domain and the heme moiety in hemoglobin binding. Binding of rHA2 with either hemin, protoporphyrin IX, or hematoporphyrin was abolished by establishing covalent linkage of the protoporphyrin propionic acid side chains to fixed amines, demonstrating specific and directed binding of rHA2 to these protoporphyrins. A monoclonal antibody which recognizes a peptide epitope within the HA2 domain was employed to demonstrate that HA2-associated hemoglobin-binding activity was expressed and released by P. gingivalis cells in a batch culture, in parallel with proteinase activity. Cysteine proteinases from P. gingivalis appear to be multidomain proteins with functions for hemagglutination, erythrocyte lysis, proteolysis, and heme binding, as demonstrated here. Detailed understanding of the biochemical pathways for heme acquisition in P. gingivalis may allow precise targeting of this critical metabolic aspect for periodontal disease prevention. PMID- 10368155 TI - An intracellular iron chelator pleiotropically suppresses enzymatic and growth defects of superoxide dismutase-deficient Escherichia coli. AB - Mutants of Escherichia coli that lack cytoplasmic superoxide dismutase (SOD) exhibit auxotrophies for sulfur-containing, branched-chain, and aromatic amino acids and cannot catabolize nonfermentable carbon sources. A secondary-site mutation substantially relieved all of these growth defects. The requirement for fermentable carbon and the branched-chain auxotrophy occur because superoxide (O2 ) leaches iron from the [4Fe-4S] clusters of a family of dehydratases, thereby inactivating them; the suppression of these phenotypes was mediated by the restoration of activity to these dehydratases, evidently without changing the intracellular concentration of O2-. Cloning, complementation, and sequence analysis identified the suppressor mutation to be in dapD, which encodes tetrahydrodipicolinate succinylase, an enzyme involved in diaminopimelate and lysine biosynthesis. A block in dapB, which encodes dihydrodipicolinate reductase in the same pathway, conferred similar protection. Genetic analysis indicated that the protection stems from the intracellular accumulation of tetrahydro- or dihydrodipicolinate. Heterologous expression in the SOD mutants of the dipicolinate synthase of Bacillus subtilis generated dipicolinate and similarly protected them. Dipicolinates are excellent iron chelators, and their accumulation in the cell triggered derepression of the Fur regulon and a large increase in the intracellular pool of free iron, presumably as a dipicolinate chelate. A fur mutation only partially relieved the auxotrophies, indicating that Fur derepression assists but is not sufficient for suppression. It seems plausible that the abundant internal iron permits efficient reactivation of superoxide-damaged iron-sulfur clusters. This result provides circumstantial evidence that the sulfur and aromatic auxotrophies of SOD mutants are also directly or indirectly linked to iron metabolism. PMID- 10368157 TI - A role for a highly conserved protein of unknown function in regulation of Bacillus subtilis purA by the purine repressor. AB - Regulation of the purine biosynthetic gene purA was examined by using a transcriptional fusion to a luciferase reporter gene. Transcription was repressed about 10-fold by the addition of adenine and increased approximately 4.5-fold by the addition of guanosine. This regulation is mediated by a purine repressor (PurR). In a purR mutant, basal expression was increased 10-fold, and there was no further stimulation by guanosine or repression by adenine. An open reading frame, yabJ, immediately downstream from purR was found to have a role in the repression of purA by adenine. Repression by adenine was perturbed in a purR+ yabJ mutant, although guanosine regulation was retained. Mutations in the PurR PRPP binding motif abolished guanosine regulation in the yabJ mutant. Thus, PRPP appears to be required for upregulation by guanosine. The amino acid sequence of YabJ is homologous to the YER057c/YjgF protein family of unknown function. PMID- 10368156 TI - Construction and initial characterization of Escherichia coli strains with few or no intact chromosomal rRNA operons. AB - The Escherichia coli genome carries seven rRNA (rrn) operons, each containing three rRNA genes. The presence of multiple operons has been an obstacle to many studies of rRNA because the effect of mutations in one operon is diluted by the six remaining wild-type copies. To create a tool useful for manipulating rRNA, we sequentially inactivated from one to all seven of these operons with deletions spanning the 16S and 23S rRNA genes. In the final strain, carrying no intact rRNA operon on the chromosome, rRNA molecules were expressed from a multicopy plasmid containing a single rRNA operon (prrn). Characterization of these rrn deletion strains revealed that deletion of two operons was required to observe a reduction in the growth rate and rRNA/protein ratio. When the number of deletions was extended from three to six, the decrease in the growth rate was slightly more than the decrease in the rRNA/protein ratio, suggesting that ribosome efficiency was reduced. This reduction was most pronounced in the Delta7 prrn strain, in which the growth rate, unlike the rRNA/protein ratio, was not completely restored to wild-type levels by a cloned rRNA operon. The decreases in growth rate and rRNA/protein ratio were surprisingly moderate in the rrn deletion strains; the presence of even a single operon on the chromosome was able to produce as much as 56% of wild-type levels of rRNA. We discuss possible applications of these strains in rRNA studies. PMID- 10368158 TI - Analysis of quorum-sensing-dependent control of rhizosphere-expressed (rhi) genes in Rhizobium leguminosarum bv. viciae. AB - The rhi genes of Rhizobium leguminosarum biovar viciae are expressed in the rhizosphere and play a role in the interaction with legumes, such as the pea. Previously (K. M. Gray, J. P. Pearson, J. A. Downie, B. E. A. Boboye, and E. P. Greenberg, J. Bacteriol. 178:372-376, 1996) the rhiABC operon had been shown to be regulated by RhiR and to be induced by added N-(3-hydroxy-7-cis-tetradecenoyl) L-homoserine lactone (3OH, C14:1-HSL). Mutagenesis of a cosmid carrying the rhiABC and rhiR gene region identified a gene (rhiI) that affects the level of rhiA expression. Mutation of rhiI slightly increased the number of nodules formed on the pea. The rhiI gene is (like rhiA) regulated by rhiR in a cell density dependent manner. RhiI is similar to LuxI and other proteins involved in the synthesis of N-acyl-homoserine lactones (AHLs). Chemical analyses of spent culture supernatants demonstrated that RhiI produces N-(hexanoyl)-L-homoserine lactone (C6-HSL) and N-(octanoyl)-L-homoserine lactone (C8-HSL). Both of these AHLs induced rhiA-lacZ and rhiI-lacZ expression on plasmids introduced into an Agrobacterium strain that produces no AHLs, showing that rhiI is positively regulated by autoinduction. However, in this system no induction of rhiA or rhiI with 3OH,C14:1-HSL was observed. Analysis of the spent culture supernatant of the wild-type R. leguminosarum bv. viciae revealed that at least seven different AHLs are made. Mutation of rhiI decreased the amounts of C6-HSL and C8-HSL but did not block their formation, and in this background the rhiI mutation did not significantly affect the expression levels of the rhiI gene or rhiABC genes or the accumulation of RhiA protein. These observations suggest that there are additional loci involved in AHL production in R. leguminosarum bv. viciae and that they affect rhiI and rhiABC expression. We postulate that the previously observed induction of rhiA by 3OH,C14:1-HSL may be due to an indirect effect caused by induction of other AHL production loci. PMID- 10368159 TI - relA is required for actinomycin production in Streptomyces antibioticus. AB - The relA gene from Streptomyces antibioticus has been cloned and sequenced. The gene encodes a protein with an Mr of 93,653, which is 91% identical to the corresponding protein from Streptomyces coelicolor. Disruption of S. antibioticus relA produces a strain which grows significantly more slowly on actinomycin production medium than the wild type or a disruptant to which the intact relA gene was restored. Moreover, the disruptant was unable to accumulate ppGpp to the levels observed during the normal course of growth and actinomycin production in the wild type. The strain containing the disrupted relA gene did not produce actinomycin and contained significantly lower levels of the enzyme phenoxazinone synthase than the wild-type strain. Actinomycin synthetase I, a key enzyme in the actinomycin biosynthetic pathway, was undetectable in the relA disruptant. Growth of the disruptant on low-phosphate medium did not restore actinomycin production. PMID- 10368160 TI - AGT1, encoding an alpha-glucoside transporter involved in uptake and intracellular accumulation of trehalose in Saccharomyces cerevisiae. AB - The trehalose content in Saccharomyces cerevisiae can be significantly manipulated by including trehalose at an appropriate level in the growth medium. Its uptake is largely dependent on the expression of AGT1, which encodes an alpha glucoside transporter. The trehalose found in a tps1 mutant of trehalose synthase may therefore largely reflect its uptake from the enriched medium that was employed. PMID- 10368162 TI - Biochemical and molecular characterization of the Bacillus subtilis acetoin catabolic pathway. AB - A recent study indicated that Bacillus subtilis catabolizes acetoin by enzymes encoded by the acu gene cluster (F. J. Grundy, D. A. Waters, T. Y. Takova, and T. M. Henkin, Mol. Microbiol. 10:259-271, 1993) that are completely different from those in the multicomponent acetoin dehydrogenase enzyme system (AoDH ES) encoded by aco gene clusters found before in all other bacteria capable of utilizing acetoin as the sole carbon source for growth. By hybridization with a DNA probe covering acoA and acoB of the AoDH ES from Clostridium magnum, genomic fragments from B. subtilis harboring acoA, acoB, acoC, acoL, and acoR homologous genes were identified, and some of them were functionally expressed in E. coli. Furthermore, acoA was inactivated in B. subtilis by disruptive mutagenesis; these mutants were impaired to express PPi-dependent AoDH E1 activity to remove acetoin from the medium and to grow with acetoin as the carbon source. Therefore, acetoin is catabolized in B. subtilis by the same mechanism as all other bacteria investigated so far, leaving the function of the previously described acu genes obscure. PMID- 10368161 TI - Role for the oxyS gene in regulation of intracellular hydrogen peroxide in Escherichia coli. AB - Intracellular hydrogen peroxide is regulated in Escherichia coli by OxyR in response to the metabolic production of H2O2. Here, we show that the untranslated oxyS RNA controlled by OxyR has a role in this regulation. The oxyS transcript appears to affect the metabolic output of H2O2 rather than the removal of H2O2 by catalases-hydroperoxidases. PMID- 10368163 TI - Identification of two new proteins in spermidine nucleoids isolated from Escherichia coli. AB - The Escherichia coli nucleoid contains DNA in a condensed but functional form. Analysis of proteins released from isolated spermidine nucleoids after treatment with DNase I reveals significant amounts of two proteins not previously detected in wild-type E. coli. Partial amino-terminal sequencing has identified them as the products of rdgC and yejK. These proteins are strongly conserved in gram negative bacteria, suggesting that they have important cellular roles. PMID- 10368164 TI - Cloning and characterization of the gene encoding Pasteurella haemolytica FnrP, a regulator of the Escherichia coli silent hemolysin sheA. AB - A Pasteurella haemolytica A1 gene was identified from a recombinant library clone that expressed hemolysis in host Escherichia coli cells. The gene, designated fnrP, had sequence identity to E. coli fnr, a global transcriptional regulator of genes required for conversion to anaerobic growth. FnrP complemented anaerobic deficiencies of a fnr-null mutant strain of E. coli and increased expression of the Fnr-dependent, anaerobic terminal reductase gene, frdA. FnrP was purified, identified by immunoblotting, and shown to be nonhemolytic. When FnrP was expressed in E. coli DeltasheA, a null mutant of the cryptic hemolysin SheA, the transformants were nonhemolytic, indicating that FnrP activates this silent hemolysin. PMID- 10368165 TI - Uptake of pyocin S3 occurs through the outer membrane ferripyoverdine type II receptor of Pseudomonas aeruginosa. AB - Pyocin S3 was found to kill exclusively Pseudomonas aeruginosa isolates producing type II pyoverdine (exemplified by strain ATCC 27853). Killing was specifically inhibited by addition of type II ferripyoverdine. All Tn5 mutants resistant to pyocin S3 were defective for pyoverdine-mediated iron uptake and failed to produce an 85-kDa iron-repressed outer membrane protein. We conclude that this protein is probably the type II ferripyoverdine receptor that is used by pyocin S3 to gain entry into the cell. PMID- 10368166 TI - Mutational analysis of the Streptococcus pneumoniae bimodular class A penicillin binding proteins. AB - One group of penicillin target enzymes, the class A high-molecular-weight penicillin-binding proteins (PBPs), are bimodular enzymes. In addition to a central penicillin-binding-transpeptidase domain, they contain an N-terminal putative glycosyltransferase domain. Mutations in the genes for each of the three Streptococcus pneumoniae class A PBPs, PBP1a, PBP1b, and PBP2a, were isolated by insertion duplication mutagenesis within the glycosyltransferase domain, documenting that their function is not essential for cellular growth in the laboratory. PBP1b PBP2a and PBP1a PBP1b double mutants could also be isolated, and both showed defects in positioning of the septum. Attempts to obtain a PBP2a PBP1a double mutant failed. All mutants with a disrupted pbp2a gene showed higher sensitivity to moenomycin, an antibiotic known to inhibit PBP-associated glycosyltransferase activity, indicating that PBP2a is the primary target for glycosyltransferase inhibitors in S. pneumoniae. PMID- 10368167 TI - Fused and overlapping rpoB and rpoC genes in Helicobacters, Campylobacters, and related bacteria. AB - The genes coding for the beta (rpoB) and beta' (rpoC) subunits of RNA polymerase are fused in the gastric pathogen Helicobacter pylori but separate in other taxonomic groups. To better understand how the unique fused structure evolved, we determined DNA sequences at and around the rpoB-rpoC junction in 10 gastric and nongastric species of Helicobacter and in members of the related genera Wolinella, Arcobacter, Sulfurospirillum, and Campylobacter. We found the fusion to be specific to Helicobacter and Wolinella genera; rpoB and rpoC overlap in the other genera. The fusion may have arisen by a frameshift mutation at the site of rpoB and rpoC overlap. Loss of good Shine-Dalgarno sequences might then have fixed the fusion in the Helicobacteraceae, even if fusion itself did not confer a selective advantage. PMID- 10368168 TI - Genotype, phenotype, and protein structure in a regulator of sporulation: effects of mutations in the spoIIAA gene of Bacillus subtilis. AB - SpoIIAA, a phosphorylatable protein, is essential to the regulation of sigmaF, the first sporulation-specific transcription factor of Bacillus subtilis. The solution structure of SpoIIAA has recently been published. Here we examine four mutant SpoIIAA proteins and correlate their properties with the phenotypes of the corresponding B. subtilis mutant strains. Two of the mutations severely disrupted the structure of the protein, a third greatly diminished the rate of its phosphorylation and abolished dephosphorylation, and the fourth left phosphorylation unaffected but reduced the rate of dephosphorylation about 10 fold. PMID- 10368170 TI - The nucleocytoplasmic continuum. Pushing the (nuclear) envelope. PMID- 10368171 TI - Trends in plant cell cycle research. PMID- 10368169 TI - Activation of Escherichia coli leuV transcription by FIS. AB - The transcription factor FIS has been implicated in the regulation of several stable RNA promoters, including that for the major tRNALeu species in Escherichia coli, tRNA1Leu. However, no evidence for direct involvement of FIS in tRNA1Leu expression has been reported. We show here that FIS binds to a site upstream of the leuV promoter (centered at -71) and that it directly stimulates leuV transcription in vitro. A mutation in the FIS binding site reduces transcription from a leuV promoter in strains containing FIS but has no effect on transcription in strains lacking FIS, indicating that FIS contributes to leuV expression in vivo. We also find that RNA polymerase forms an unusual heparin-sensitive complex with the leuV promoter, having a downstream protection boundary of approximately 7, and that the first two nucleotides of the transcript, GTP and UTP, are required for formation of a heparin-stable complex that extends downstream of the transcription start site. These studies have implications for the regulation of leuV transcription. PMID- 10368172 TI - Isovariant dynamics expand and buffer the responses of complex systems: the diverse plant actin gene family. PMID- 10368173 TI - Interactions among APETALA1, LEAFY, and TERMINAL FLOWER1 specify meristem fate. AB - Upon floral induction, the primary shoot meristem of an Arabidopsis plant begins to produce flower meristems rather than leaf primordia on its flanks. Assignment of floral fate to lateral meristems is primarily due to the cooperative activity of the flower meristem identity genes LEAFY (LFY), APETALA1 (AP1), and CAULIFLOWER. We present evidence here that AP1 expression in lateral meristems is activated by at least two independent pathways, one of which is regulated by LFY. In lfy mutants, the onset of AP1 expression is delayed, indicating that LFY is formally a positive regulator of AP1. We have found that AP1, in turn, can positively regulate LFY, because LFY is expressed prematurely in the converted floral meristems of plants constitutively expressing AP1. Shoot meristems maintain an identity distinct from that of flower meristems, in part through the action of genes such as TERMINAL FLOWER1 (TFL1), which bars AP1 and LFY expression from the influorescence shoot meristem. We show here that this negative regulation can be mutual because TFL1 expression is downregulated in plants constitutively expressing AP1. Therefore, the normally sharp phase transition between the production of leaves with associated shoots and formation of the flowers, which occurs upon floral induction, is promoted by positive feedback interactions between LFY and AP1, together with negative interactions of these two genes with TFL1. PMID- 10368175 TI - Hormonally regulated programmed cell death in barley aleurone cells AB - Cell death was studied in barley (cv Himalaya) aleurone cells treated with abscisic acid and gibberellin. Aleurone protoplasts incubated in abscisic acid remained viable in culture for at least 3 weeks, but exposure to gibberellin initiated a series of events that resulted in death. Between 4 and 8 days after incubation in gibberellin, >70% of all protoplasts died. Death, which occurred after cells became highly vacuolated, was manifest by an abrupt loss of plasma membrane integrity followed by rapid shrinkage of the cell corpse. Hydrolysis of DNA began before death and occurred as protoplasts ceased production of alpha amylase. DNA degradation did not result in the accumulation of discrete low molecular weight fragments. DNA degradation and cell death were prevented by LY83583, an inhibitor of gibberellin signaling in barley aleurone. We conclude that cell death in aleurone cells is hormonally regulated and is the final step of a developmental program that promotes successful seedling establishment. PMID- 10368174 TI - The Arabidopsis dwarf mutant shi exhibits reduced gibberellin responses conferred by overexpression of a new putative zinc finger protein. AB - shi (for short internodes), a semidominant dwarfing mutation of Arabidopsis caused by a transposon insertion, confers a phenotype typical of mutants defective in the biosynthesis of gibberellin (GA). However, the application of GA does not correct the dwarf phenotype of shi plants, suggesting that shi is defective in the perception of or in the response to GA. In agreement with this observation, the level of active GAs was elevated in shi plants, which is the result expected when feedback control of GA biosynthesis is reduced. Cloning of the SHI gene revealed that in shi, the transposon is inserted into the untranslated leader so that a cauliflower mosaic virus 35S promoter in the transposon reads out toward the SHI open reading frame. This result, together with mRNA analysis, suggests that the phenotype of the shi mutant is a result of overexpression of the SHI open reading frame. The predicted amino acid sequence of SHI has acidic and glutamine-rich stretches and shows sequence similarity over a putative zinc finger region to three presumptive Arabidopsis proteins. This suggests that SHI may act as a negative regulator of GA responses through transcriptional control. PMID- 10368177 TI - Silencing gene expression of the ethylene-forming enzyme results in a reversible inhibition of ovule development in transgenic tobacco plants AB - To study the role of ethylene in plant reproduction, we constructed transgenic tobacco plants in which the expression of a pistil-specific gene coding for the ethylene-forming enzyme 1-aminocyclopropane-1-carboxylate oxidase was inhibited. Flowers from transgenic plants showed female sterility due to an arrest in ovule development. Megasporogenesis did not occur, and ovules did not reach maturity. When pollinated, pollen tubes were able to reach the ovary but did not penetrate into the immature ovule in transgenic plants. Flower treatment with an ethylene source resulted in a functional recovery of ovule development and restored guidance of the pollen tube tip into the ovule micropyle that resulted in seed set. The recovery was abolished if inhibitors of ethylene action were present. These results demonstrate that the plant hormone ethylene is required during the very early stages of female sporogenesis and ultimately to enable fertilization. PMID- 10368176 TI - Transgenic tobacco plants expressing the Drosophila Polycomb (Pc) chromodomain show developmental alterations: possible role of Pc chromodomain proteins in chromatin-mediated gene regulation in plants. AB - The chromodomain of the Drosophila Polycomb (Pc) protein has been introduced into tobacco nuclei to determine its location in the nucleus and its effect on plant development. Pc is a repressor of homeotic Drosophila genes that shares a well conserved, although not identical, chromodomain with a structural heterochromatin component, Heterochromatin Protein 1. The chromodomains might therefore play a common role in chromatin repression. An analysis of transgenic plants expressing the Pc chromodomain, which was linked to the green fluorescent protein, suggested that the Pc chromodomain has distinct target regions in the plant genome. Transgenic plants expressing the Pc chromodomain had phenotypic abnormalities in their leaves and flowers, indicating a disruption in development. In axillary shoot buds of plants displaying altered leaf phenotypes, enhanced expression of a homeodomain gene, which is downregulated in wild-type leaves, was found. In Drosophila, Pc has been shown to possess distinct chromosome binding activity and to be involved in the regulation of development-specific genes. Our results support the assumptions that the heterologous chromodomain affects related functions in Drosophila and in plants, and that chromatin modification mechanisms are involved in the regulation of certain plant genes, in a manner similar to chromatin-mediated gene regulation in Drosophila. PMID- 10368178 TI - Leaf senescence is delayed in tobacco plants expressing the maize homeobox gene knotted1 under the control of a senescence-activated promoter. AB - Leaf senescence is an active process involving remobilization of nutrients from senescing leaves to other parts of the plant. Whereas senescence is accompanied by a decline in leaf cytokinin content, supplemental cytokinin delays senescence. Plants that overexpress isopentenyl transferase (ipt), a cytokinin-producing gene, or knotted1 (kn1), a homeobox gene, have many phenotypes in common. Many of these phenotypes are characteristic of altered cytokinin physiology. The effect of kn1 on leaf senescence was tested by driving its expression using the promoter of the senescence-associated gene SAG12. SAG:kn1 tobacco plants showed a marked delay in leaf senescence but otherwise developed normally. The delay in senescence was revealed by an increase in chlorophyll content in SAG:kn1 leaves relative to leaves of the control plants and by a decrease in the number of dead leaves. Senescence was also delayed in detached leaves of SAG:kn1 plants. Delayed senescence was accompanied by increased leaf cytokinin content in older leaves expressing kn1. These experiments extend the current understanding of kn1 function and suggest that in addition to mediating meristem maintenance, kn1 is capable of regulating the onset of senescence in leaves. PMID- 10368180 TI - GEG participates in the regulation of cell and organ shape during corolla and carpel development in gerbera hybrida AB - The molecular mechanisms that control organ shape during flower development are largely unknown. By using differential hybridization techniques, a cDNA designated GEG (for Gerbera hybrida homolog of the gibberellin [GA]-stimulated transcript 1 [GAST1] from tomato) was isolated from a library representing late stages of corolla development in Gerbera. GEG expression was detected in corollas and carpels, with expression spatiotemporally coinciding with flower opening. In corollas and styles, GEG expression is temporally correlated with the cessation of longitudinal cell expansion. In plants constitutively expressing GEG, reduced corolla lengths and carpels with shortened and radially expanded stylar parts were found, with concomitant reduction of longitudinal cell expansion in these organs. In addition, in styles, an increase in radial cell expansion was detected. Taken together, these observations indicate a regulatory role for the GEG gene product in determining the shape of the corolla and carpel. The deduced amino acid sequence of the GEG gene product shares high similarity with previously characterized putative cell wall proteins encoded by GA-inducible genes, namely, GAST1, GIP (for GA-induced gene of petunia), and the GASA (for GA stimulated in Arabidopsis) gene family. Our studies suggest that GEG, the expression of which can also be induced by application of GA3, plays a role in phytohormone-mediated cell expansion. PMID- 10368179 TI - Structure of a plant cell wall fragment complexed to pectate lyase C. AB - The three-dimensional structure of a complex between the pectate lyase C (PelC) R218K mutant and a plant cell wall fragment has been determined by x-ray diffraction techniques to a resolution of 2.2 A and refined to a crystallographic R factor of 18.6%. The oligosaccharide substrate, alpha-D-GalpA-([1-->4]-alpha-D GalpA)3-(1-->4)-D-GalpA , is composed of five galacturonopyranose units (D-GalpA) linked by alpha-(1-->4) glycosidic bonds. PelC is secreted by the plant pathogen Erwinia chrysanthemi and degrades the pectate component of plant cell walls in soft rot diseases. The substrate has been trapped in crystals by using the inactive R218K mutant. Four of the five saccharide units of the substrate are well ordered and represent an atomic view of the pectate component in plant cell walls. The conformation of the pectate fragment is a mix of 21 and 31 right handed helices. The substrate binds in a cleft, interacting primarily with positively charged groups: either lysine or arginine amino acids on PelC or the four Ca2+ ions found in the complex. The observed protein-oligosaccharide interactions provide a functional explanation for many of the invariant and conserved amino acids in the pectate lyase family of proteins. Because the R218K PelC-galacturonopentaose complex represents an intermediate in the reaction pathway, the structure also reveals important details regarding the enzymatic mechanism. Notably, the results suggest that an arginine, which is invariant in the pectate lyase superfamily, is the amino acid that initiates proton abstraction during the beta elimination cleavage of polygalacturonic acid. PMID- 10368181 TI - Generation of a spacing pattern: the role of triptychon in trichome patterning in Arabidopsis. AB - Trichomes in Arabidopsis are single-celled hairs that exhibit a regular spacing pattern. Here, the role of TRIPTYCHON (TRY) in the generation of this spacing pattern is studied. By using genetic mosaics, we demonstrate that the formation of trichome clusters in try mutants is not correlated with cell lineage, indicating that TRY is required to single out trichome cells in a process involving cellular interactions. The genetic interactions of TRY, GLABRA1 (GL1), and TRANSPARENT TESTA GLABRA (T TG) in trichome patterning are assessed by determining the cluster frequency in various genetic combinations. It is shown that TRY acts as a negative regulator of GL1- and TTG-dependent pathways. Furthermore, it is demonstrated that trichome initiation in ttg-1, a strong ttg allele, is rescued almost to wild-type levels in a try background in which GL1 is expressed under the control of the cauliflower mosaic virus 35S promoter, indicating that T TG acts upstream of GL1 and TRY. These findings are incorporated into a model to explain the generation of a trichome spacing pattern from a homogeneous population of epidermal cells. PMID- 10368182 TI - Matrix attachment region binding protein MFP1 is localized in discrete domains at the nuclear envelope. AB - Recently, it has been suggested that nuclear processes, such as replication, transcription, and splicing, are spatially organized and associated with a nuclear framework called the nuclear matrix, a structure of unknown molecular composition. It has been shown that chromatin is attached to the nuclear matrix via specific DNA fragments called matrix attachment regions (MARs). We have begun to dissect the plant nuclear matrix by isolating a DNA binding protein with specific affinity for MARs. Here, it is shown that MAR binding filament-like protein 1 (MFP1) is associated with specklelike structures at the nuclear periphery that are part of isolated nuclei and the nuclear matrix. A predicted N terminal transmembrane domain is necessary for the specific targeting of MFP1 to the speckles, indicating an association with the nuclear envelope-endoplasmic reticulum continuum. In addition, it is shown that a marker protein for plant microtubule organizing centers, which has been shown to be localized on the outside of the plant nuclear envelope, is also part of the nuclear matrix. These findings indicate a close and previously undescribed connection in plants between the nuclear envelope and the internal nuclear matrix, and they suggest a function for MFP1 in attaching chromatin to specific sites at the nuclear periphery. PMID- 10368183 TI - Effect of pectin methylesterase gene expression on pea root development. AB - Expression of an inducible gene with sequences common to genes encoding pectin methylesterase (PME) was found to be tightly correlated, both spatially and temporally, with border cell separation in pea root caps. Partial inhibition of the gene's expression by antisense mRNA in transgenic pea hairy roots prevented the normal separation of root border cells from the root tip into the external environment. This phenotype was correlated with an increase in extracellular pH, reduced root elongation, and altered cellular morphology. The translation product of the gene exhibited PME activity in vitro. These results are consistent with the long-standing hypothesis that the demethylation of pectin by PME plays a key role in cell wall metabolism. PMID- 10368184 TI - ATP binding cassette modulators control abscisic acid-regulated slow anion channels in guard cells AB - In animal cells, ATP binding cassette (ABC) proteins are a large family of transporters that includes the sulfonylurea receptor and the cystic fibrosis transmembrane conductance regulator (CFTR). These two ABC proteins possess an ion channel activity and bind specific sulfonylureas, such as glibenclamide, but homologs have not been identified in plant cells. We recently have shown that there is an ABC protein in guard cells that is involved in the control of stomatal movements and guard cell outward K+ current. Because the CFTR, a chloride channel, is sensitive to glibenclamide and able to interact with K+ channels, we investigated its presence in guard cells. Potent CFTR inhibitors, such as glibenclamide and diphenylamine-2-carboxylic acid, triggered stomatal opening in darkness. The guard cell protoplast slow anion current that was recorded using the whole-cell patch-clamp technique was inhibited rapidly by glibenclamide in a dose-dependent manner; the concentration producing half maximum inhibition was at 3 &mgr;M. Potassium channel openers, which bind to and act through the sulfonylurea receptor in animal cells, completely suppressed the stomatal opening induced by glibenclamide and recovered the glibenclamide inhibited slow anion current. Abscisic acid is known to regulate slow anion channels and in our study was able to relieve glibenclamide inhibition of slow anion current. Moreover, in epidermal strip bioassays, the stomatal closure triggered by Ca2+ or abscisic acid was reversed by glibenclamide. These results suggest that the slow anion channel is an ABC protein or is tightly controlled by such a protein that interacts with the abscisic acid signal transduction pathway in guard cells. PMID- 10368185 TI - Phytochelatin synthase genes from Arabidopsis and the yeast Schizosaccharomyces pombe. AB - Phytochelatins (PCs), a family of heavy metal-inducible peptides important in the detoxification of heavy metals, have been identified in plants and some microorganisms, including Schizosaccharomyces pombe, but not in animals. PCs are synthesized enzymatically from glutathione (GSH) by PC synthase in the presence of heavy metal ions. In Arabidopsis, the CAD1 gene, identified by using Cd sensitive, PC-deficient cad1 mutants, has been proposed to encode PC synthase. Using a positional cloning strategy, we have isolated the CAD1 gene. Database searches identified a homologous gene in S. pombe, and a mutant with a targeted deletion of this gene was also Cd sensitive and PC deficient. Extracts of Escherichia coli cells expressing a CAD1 cDNA or the S. pombe gene catalyzing GSH dependent, heavy metal-activated synthesis of PCs in vitro demonstrated that both genes encode PC synthase activity. Both enzymes were activated by a range of metal ions. In contrast, reverse transcription-polymerase chain reaction experiments showed that expression of the CAD1 mRNA is not influenced by the presence of Cd. A comparison of the two predicted amino acid sequences revealed a highly conserved N-terminal region, which is presumed to be the catalytic domain, and a variable C-terminal region containing multiple Cys residues, which is proposed to be involved in activation of the enzyme by metal ions. Interestingly, a similar gene was identified in the nematode, Caenorhabditis elegans, suggesting that PCs may also be expressed in some animal species. PMID- 10368186 TI - A chloroplast-targeted heat shock protein 70 (HSP70) contributes to the photoprotection and repair of photosystem II during and after photoinhibition. AB - Dark-grown Chlamydomonas reinhardtii cultures that were illuminated at low fluence rates before exposure to high-light conditions exhibited a faster rate of recovery from photoinhibition than did dark-grown cells that were directly exposed to photoinhibitory conditions. This pretreatment has been shown to induce the expression of several nuclear heat shock protein 70 (HSP70) genes, including HSP70B, encoding a chloroplast-localized chaperone. To investigate a possible role of plastidic HSP70B in photoprotection and repair of photosystem II, which is the major target of photoinhibition, we have constructed strains overexpressing or underexpressing HSP70B. The effect of light stress on photosystem II in nuclear transformants harboring HSP70B in the sense or antisense orientation was monitored by measuring variable fluorescence, flash induced charge separation, and relative amounts of various photosystem II polypeptides. Underexpression of HSP70B caused an increased light sensitivity of photosystem II, whereas overexpression of HSP70B had a protective effect. Furthermore, the reactivation of photosystem II after photoinhibition was enhanced in the HSP70B-overexpressing strain when compared with the wild type, both in the presence or absence of synthesis of chloroplast-encoded proteins. Therefore, HSP70B may participate in vivo both in the molecular protection of the photosystem II reaction centers during photoinhibition and in the process of photosystem II repair. PMID- 10368187 TI - Sac3, an Snf1-like serine/threonine kinase that positively and negatively regulates the responses of Chlamydomonas to sulfur limitation. AB - The Sac3 gene product of Chlamydomonas positively and negatively regulates the responses of the cell to sulfur limitation. In wild-type cells, arylsulfatase activity is detected only during sulfur limitation. The sac3 mutant expresses arylsulfatase activity even when grown in nutrient-replete medium, which suggests that the Sac3 protein has a negative effect on the induction of arylsulfatase activity. In contrast to its effect on arylsulfatase activity, Sac3 positively regulates the high-affinity sulfate transport system-the sac3 mutant is unable to fully induce high-affinity sulfate transport during sulfur limitation. We have complemented the sac3 mutant and cloned a cDNA copy of the Sac3 gene. The deduced amino acid sequence of the Sac3 gene product is similar to the catalytic domain of the yeast Snf1 family of serine/threonine kinases and is therefore classified as a Snf1-related kinase (SnRK). Specifically, Sac3 falls within the SnRK2 subfamily of kinases from vascular plants. In addition to the 11 subdomains common to Snf1-like serine/threonine kinases, Sac3 and the plant kinases have two additional subdomains and a highly acidic C-terminal region. The role of Sac3 in the signal transduction system that regulates the responses of Chlamydomonas to sulfur limitation is discussed. PMID- 10368188 TI - MN/CA9 gene expression as a potential biomarker in renal cell carcinoma. AB - OBJECTIVE: To compare the levels of MN/CA9 expression with clinicopathological variables in renal cell carcinoma (RCC), and thus evaluate MN/CA9 expression as a possible biomarker for RCC. MATERIALS AND METHODS: The level of expression of MN/CA9 was evaluated in 76 surgically obtained tissue samples (49 from RCC, 22 from normal kidney and five from oncocytoma) using semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis. In RCC, MN/CA9 expression was compared with stage, grade and cell type. RESULTS: MN/CA9 was expressed in 42 of 49 (86%) RCC samples, but in only two of 22 (9%) normal kidney and none of five oncocytoma samples. Levels of MN/CA9 expression were significantly higher in tumours consisting only of clear cells than in those containing other cell types (P=0.0189), and MN/CA9 was expressed in 34 of 37 (92%) RCC samples consisting only of clear cells. Tumours of low clinical stage showed a striking increase in MN/CA9 expression, and high MN/CA9 expression was associated with a good patient outcome. CONCLUSION: These results suggest that MN/CA9 expression is a potential diagnostic biomarker of RCC, especially the clear-cell type, and can be targeted using molecular biological techniques. PMID- 10368190 TI - Buccal mucosal grafts in the treatment of ureteric lesions. AB - OBJECTIVE: To devise a procedure capable of curing complicated ureteric strictures and replacing segments of lost ureter, without the long-term infective complications of bowel interposition or the surgical magnitude of autotransplantation. PATIENTS AND METHODS: Four patients with complicated strictures and one with segmental ureteric loss were treated by buccal mucosal patch grafts and an omental wrap. One patient with segmental ureteric loss was treated by interposition of a tubularized buccal mucosal graft. RESULTS: Ureteric patency was established and maintained in all patients, there were no complications and urine was sterile in all patients at follow-up. CONCLUSION: In a few patients, buccal mucosal patch graft repair has proved capable of maintaining patency and good urinary drainage in patients with complicated ureteric strictures. Segmental ureteric loss has been replaced in one patient by a patch graft and in another by tubularized graft interposition. PMID- 10368189 TI - Outpatient flexible cystoscope-assisted insertion of ureteric catheters and ureteric stents. AB - OBJECTIVE: To examine the feasibility of inserting ureteric catheters (before retrograde ureterography) and JJ ureteric stents (both traditionally performed under general anaesthesia on inpatients) using local anaesthesia in an outpatient setting and with no patient selection bias, thus providing procedures to ease the demand on inpatient lists. PATIENTS AND METHODS: All patients presenting with an appropriate clinical indication for either of the procedures underwent insertion during a scheduled flexible cystoscopy session in the outpatient department; information was collected on a standard proforma. Antibiotic prophylaxis and a nonsteroidal analgesic were administered 30 min before the procedure. RESULTS: To date, 20 patients (seven men, age range 32-74 years, and 13 women, age range 23 86 years) have undergone one of the two procedures. Fourteen patients had attempted retrograde catheter insertion, with success in 12, and six other patients underwent attempted JJ ureteric stent insertion, with success in five. The three failures were caused by an inability to see the relevant ureteric orifice. The mean duration of each procedure was 11 min; 14 of the 17 patients who had a successful procedure had no significant pain or discomfort, while the remaining three experienced significant pain and discomfort. There were no infective complications. CONCLUSION: These two procedures are suitable for the outpatient/day-case situation, and are well tolerated and accepted by most patients. PMID- 10368191 TI - A new thermo-expandable shape-memory nickel-titanium alloy stent for the management of ureteric strictures. AB - OBJECTIVE: To assess the ease on insertion, patient tolerance, undesirable side effects, degree of encrustation and duration of upper tract decompression with a new thermo-expandable shape memory alloy ureteric stent. PATIENTS AND METHODS: From November 1996 to October 1998, 15 patients with ureteric strictures were treated with a new nickel-titanium shape-memory alloy stent, the Memokath 051 (Engineers & Doctors A/S, Hornbaek, Denmark). A total of 22 insertions were carried out. Ureteric obstruction was caused by recurrent colorectal carcinoma in four patients; two patients each with transitional cell carcinoma of the bladder, iatrogenic injury or ischaemia at the uretero-ileal anastomosis; and one patient each with metastatic lymph nodes from prostatic carcinoma, radiation-induced fibrosis, pelvi-ureteric junction obstruction, metastatic carcinoma of the vagina and extra-luminal endometriosis. The stent has a shaft diameter of 9 F and its proximal end expands to 17 F. The first three patients were treated with the original version, which expanded to 14 F. The unexpanded stent is inserted into the ureter after initial dilatation of the stricture to 12 F. The stent is expanded by injection with sterile water preheated to 50 degrees C. The procedures were carried out under a general anaesthetic and patients were allowed home the next day. The follow-up protocol included initial intravenous urography (IVU) at 6 weeks, with assessment of a mid-stream urine sample and renal function tests. These were repeated at 3-monthly intervals. Isotopic renography was performed when indicated. RESULTS: The mean (range) follow-up was 10.6 (2-21) months; there was complete relief of upper tract obstruction in all patients. No stent-related symptoms, e.g. pain, sepsis, haematuria or frequency, were noted and no encrustation has occurred so far. The stent migrated in the first three patients with the original smaller diameter of stent but decompression of the upper tracts was maintained. None of the modified wider stents have migrated. The return of peristalsis in the proximal ureter was detected during IVU. There was no apparent endothelial growth through the stent material and no re-admissions for stent-related complications. CONCLUSION: Early experience with this new stent is very encouraging. All patients have maintained satisfactory decompression of their upper tracts with no need for repeated hospitalization for stent changes. There have been no untoward side-effects so far. This stent appears to have a valuable place in the long-term management of ureteric strictures; it is probably most suited for malignant ureteric obstruction. It should be considered in the management of selected benign strictures that require long-term JJ stenting. PMID- 10368192 TI - Urinary incontinence in Northern Ireland: a prevalence study. AB - OBJECTIVE: To determine the prevalence of urinary incontinence in a Northern Ireland community drawn from four neighbouring geographical areas and to assess factors predisposing to the development of urinary incontinence. SUBJECTS AND METHODS: A three-page self-administered postal questionnaire was sent to 1050 women (age range 35-74 years), recruited randomly from a target population of 43 829 women. The main survey was preceded by a pilot survey. Respondents and those not responding were compared. RESULTS: The overall response rate was 65.6% (689/1050); there was no significant difference between respondents and those not responding. Two-hundred and thirty-one women (33.5%) reported incontinence 'sometimes' and 161 (23.4%) 'often'. Of those who had urinary incontinence, sanitary protection was required by 21.7% (85/392). This equates to 12.3% (85) of the total study population. Age (chi2=20.34; P<0.001) and parity (Mann-Whitney U test, P< 0.001) were associated with urinary incontinence, with a higher proportion of women aged 45-54 years having urinary incontinence. The menopause and postnatal pelvic floor exercises were not associated with urinary incontinence. Overall 19.9% (78/392) of women with urinary incontinence had consulted their general practitioner. Of those who required sanitary protection, 40% (34/85) had consulted their doctor. CONCLUSION: Urinary incontinence is common; it is sufficiently severe to require sanitary protection in 12% of women aged 35-74 years in a Northern Ireland community. PMID- 10368193 TI - A comparison of prelubricated hydrophilic and non-hydrophilic polyvinyl chloride catheters for urethral catheterization. AB - OBJECTIVE: To evaluate whether patients performing clean intermittent self catheterization (CISC) for a short period preferred a prelubricated, hydrophilic, disposable polyvinyl chloride (PVC) catheter or a non-hydrophilic PVC catheter which could be used several times and that had to be lubricated by the patient. PATIENTS AND METHODS: In a prospective cross-over study, 32 patients used each type of catheter for 3 weeks. After each 3-week period, the patients completed a questionnaire to assess comfort and preference, and urine specimens were obtained for culture. RESULTS: There was no significant difference between the groups in the frequency of CISC, discomfort when used, opinion on handling the catheters, preference toward one of the catheters, or of infection. CONCLUSION: Non hydrophilic PVC catheters may be used safely and with no discomfort to the patient. In addition it may be possible for the healthcare system to save money, as the non-hydrophilic PVC catheters are much cheaper. PMID- 10368194 TI - The physicochemical basis of urinary catheter encrustation. AB - OBJECTIVES: To determine the relationship between urinary pH and Ca2+ solubility in urine samples from patients who experienced either frequent ('blockers') or infrequent ('nonblockers') catheter blockage by crystalline deposits of divalent cation salts. MATERIALS AND METHODS: Fresh urine samples from 'blockers' and 'nonblockers' were collected and the ionic calcium concentration ([Ca2+ ]) measured using a Ca2+-selective electrode whilst the urinary pH was increased in 0.25 increments between 4.75 and 9.00. The pH at which crystallization occurred (nucleation) was determined and crystal composition analysed. RESULTS: The mean (sd) voided urinary pH of catheter 'blockers' was significantly more alkaline than that from 'nonblockers', at 7.63 (0.64) and 5.97 (0.80), respectively (P=0. 001). The nucleation pH of catheter 'blockers' was significantly more acid than in 'nonblockers', at 7.43 (0.73) and 6.45 (0.65), respectively (P=0.005). Urine from 'blockers' had significantly more Ca phosphate and Mg ammonium phosphate crystals. 'Blockers' were further divided into two subsets with and without urease-based urinary tract infection; both showed a decrease in the nucleation pH. CONCLUSION: In the urine from 'nonblockers' there is a wide safety margin between voided and nucleation pHs; this margin was less in the urine from 'blockers'. This reduction in the safety margin arises partly because the voided pH in those with a urinary tract infection is more alkaline. However, the decrease in the nucleation pH also suggests that a fundamental property of urine is altered, which reduces Ca2+ solubility at more neutral pH values. The long term goal is to increase the nucleation pH of catheter 'blockers' and increase the margin of safety. PMID- 10368195 TI - A century of prostatic surgery. PMID- 10368196 TI - Effect of transurethral electrovaporization of the prostate on serum prostate specific antigen concentration. AB - OBJECTIVE: To assess the effect of transurethral electrovaporization of the prostate (TUVP) on serum prostate specific antigen (PSA) concentration. PATIENTS AND METHODS: Twenty-five men (mean age 61.7 years) with symptoms of prostatism underwent TUVP. Serum PSA levels were determined before any prostatic manipulation, and again at 1 and 24 h after TUVP. In the first 6 weeks after TUVP, serum PSA was measured every week. Prostatic size was measured by transrectal ultrasonography before and again at 6 weeks after TUVP. RESULTS: The mean serum PSA concentration was significantly higher at 1 and 24 h after TUVP (P<0.001) than before. The PSA level returned to less than the value before TUVP at 2, 3 and 4 weeks afterwards in eight (32%), 13 (52%) and 20 (80%) patients, respectively; five (20%) patients reached the baseline value 6 weeks after TUVP. The mean reduction in prostatic volume 6 weeks after TUVP was 42% and the reduction in tissue volume was significantly correlated with the decrease in serum PSA concentration at 6 weeks (P<0.05). CONCLUSION: TUVP increases serum PSA levels, the transient elevation persisting for up to 6 weeks but then declining to a stable, low PSA concentration. Therefore, it is important to wait at least 6 weeks to obtain an accurate and meaningful serum PSA level after TUVP. PMID- 10368197 TI - Changing trends in prostatic cancer. AB - OBJECTIVES: To examine the diagnosis and treatment of prostatic cancer in a population-based study, reporting incidence trends and survival, in the decade before the introduction of prostate-specific antigen (PSA) testing, and thus determine whether the overall incidence of prostatic cancer is increasing or not. PATIENTS AND METHODS: The study included all men registered as having prostatic cancer in the Yorkshire region between 1981 and 1990. The Northern and Yorkshire Cancer Registry and Information Service has an active registration policy and after notification, the information received is validated by histopathology reports and case-note review. Of the patients registered, 68% were over 70 years old at the time of diagnosis (mean age 74 years). Prostatic cancer was often diagnosed incidentally, after prostatectomy for presumed benign disease. Indications for treatment were not recorded, but most patients had treatment which was designed to control outlet bladder symptoms rather than with intent to cure cancer. RESULTS: In all, 8118 patients with prostatic cancer were registered, of whom 6587 had histological confirmation. There was a 30% increase in the age-standardized incidence of prostatic cancer during the study period (P<0.001). The mortality from prostatic cancer increased by 35% (P<0.001) and the percentage of patients known to have metastases at the time of presentation increased from 18% to 24%. These changes were seen in all age groups. The overall survival was 49% at 5 years and 34% at 10 years. CONCLUSIONS: There has been a real increase in the incidence of prostatic cancer which pre-dates the use of serum PSA testing. The percentage relative survival of patients with prostatic cancer in Yorkshire during the study period is similar to that seen in other parts of the UK, but compares badly with reported survival in other countries. PMID- 10368198 TI - Patients' tolerance of transrectal ultrasound-guided prostatic biopsy: an audit of 104 cases. AB - OBJECTIVE: To determine the frequency and severity of complications following transrectal ultrasonography (TRUS) guided prostatic biopsy, and of pain during the procedure. PATIENTS AND METHODS: The study included 129 men undergoing TRUS guided prostatic biopsy who were asked to complete a questionnaire about pain and complications one week after biopsy. RESULTS: Of the 104 men who completed the questionnaire, 24% found the procedure moderately to extremely painful and 19% felt that they had had significant complications afterward, the commonest of these being painful or difficult voiding (13%) and haematuria (11%). Systemic symptoms of fever or 'sweats' occurred in 6%, with a diagnosis of septicaemia in three men, despite antibiotic prophylaxis. However, acute urinary retention occurred in only one man. Of all patients, 20% saw their general practitioner within a week, all of whom were prescribed antibiotics in addition to those given prophylactically in hospital. CONCLUSION: TRUS-guided biopsy is often a painful experience for patients and is commonly associated with complications, particularly voiding difficulties. Of particular concern were the three patients with septicaemia, and that one in five men felt sufficiently unwell to visit their doctor within a week of the procedure. PMID- 10368199 TI - A comparison of transrectal ultrasonography and endorectal magnetic resonance imaging in the local staging of prostatic carcinoma. AB - OBJECTIVE: To compare the staging accuracy of transrectal ultrasonography (TRUS) and endorectal magnetic resonance imaging (eMRI) for organ-confined prostatic carcinoma. PATIENTS AND METHODS: Twenty-five patients with clinically confined prostatic adenocarcinoma were evaluated to be candidates for radical prostatectomy. All underwent TRUS and eMRI before surgery. Imaging findings evaluated prospectively in each patient were extracapsular extension (ECE), seminal vesicle invasion (SVI) and the site of involvement. The results of the imaging techniques were compared with the histopathological findings. As two patients with metastatic lymph nodes (detected on frozen-section examination during surgery) were spared radical prostatectomy, the final evaluation included 23 patients. RESULTS: Endorectal coil MRI was more sensitive than TRUS for detecting both ECE, SVI and the site of ECE involvement in organ-confined prostatic carcinoma. TRUS was more accurate than eMRI for detecting the site of SVI involvement. However, the overall staging accuracy rates for both imaging modalities were equal. CONCLUSIONS: Neither TRUS nor eMRI was significantly better than the other for determining the local extent of prostatic carcinoma. Therefore, TRUS should be the study of choice until MRI technology improves sufficiently in the preoperative staging of localized prostate cancer. PMID- 10368200 TI - Endocrine effects, efficacy and tolerability of a 10.8-mg depot formulation of goserelin acetate administered every 13 weeks to patients with advanced prostate cancer. AB - OBJECTIVE: To determine the endocrine effects, efficacy and tolerability of a 10.8-mg depot formulation of Zoladextrade mark (goserelin acetate, Zeneca Pharmaceuticals, Wilmington, Delaware, USA), a luteinizing hormone-releasing hormone agonist analogue, when administration was extended from every 12 weeks to every 13 weeks in patients with advanced prostate cancer. PATIENTS AND METHODS: Between July 1995 and May 1996, 59 patients with either locally advanced (T3 or T4) or metastatic prostate cancer were enrolled in an open-label, multicentre trial. Primary efficacy endpoints were testosterone measurements, and assessments of prostate specific antigen (PSA) response, subjective and objective response. Quality of life (QoL) was a secondary efficacy endpoint. RESULTS: Mean testosterone concentrations decreased to < 0.3 microgram/L by week 4 and remained so for the duration of treatment. There were no statistically significant differences in mean testosterone levels between weeks 12 and 13, or weeks 25 and 26. Serum testosterone suppression was adequate in all 58 evaluable patients at week 13, and 51 of 52 (98%) patients at week 26. Of the 58 evaluable patients, 52 (90%) had a PSA response. A subjective response was recorded for six of 11 evaluable patients. Of 58 patients evaluable for objective response, 46 (79%) had a partial response, three (5%) had stable disease and nine (16%) had objective progression. Except for a significant (P=0.014) decrease in overall sexual interest, QoL was unchanged during therapy. The most common side-effects, regardless of causality, were hot flushes (67%), pain (31%) and pelvic pain (22%). Mild injection-site complaints occurred with only three of 221 (1.4%) depot injections. CONCLUSIONS: Zoladextrade mark 10.8-mg depot, administered every 13 weeks to patients with advanced prostatic cancer, is well tolerated, provides adequate suppression of serum testosterone and produces PSA, subjective and objective responses. PMID- 10368201 TI - A circumcision service for religious reasons. AB - OBJECTIVE: To review the results of a pioneering service introduced under National Health Service (NHS) cover, providing circumcision on religious grounds. SUBJECTS AND METHODS: The service was offered to all male babies aged 6-14 weeks. Circumcision was carried out using the Plastibell technique under 1% lignocaine ring block/penile block anaesthesia. Between July 1996 and August 1998, 168 circumcisions were performed and assessed postoperatively by telephone enquiry, visiting at home by the nurse and review at the hospital if and when required. RESULTS: Of the 168 babies, 31 required some follow-up care. In six babies the separated ring tracked back onto the shaft of the penis and had to be removed using a ring-cutter. Ten babies were referred for incomplete separation of the Plastibell ring; only two required removal, the rest of the rings detaching spontaneously within a few days. Two babies were reviewed for early separation of the ring and seven were reviewed for bleeding within 24 h of the operation. Two of these were immediately after circumcision, when the bleeding was seen to be from the torn frenulum. They were converted to a more formal circumcision without the Plastibell under the same anaesthesia. Surveillance with no intervention was required in the others, except one who needed a compression dressing. One baby had apyrexia, requiring antibiotics. Four babies had suspected wound infection but none was clinically significant. One parent remained dissatisfied and complained about the inadequate removal of the foreskin. CONCLUSION: The aim of the service was to provide safe circumcision requested on religious grounds. The technique was simple and easily learned by nurses. Parents have generally been very satisfied and the complication rate was acceptable, with most problems occurring early in the series, suggesting an improvement with experience. PMID- 10368202 TI - Topical vasoactive cream in the treatment of erectile failure: a prospective, randomized placebo-controlled trial. AB - OBJECTIVE: To repeat a previous study on the use of a topical treatment for erectile failure using a vasoactive cream. PATIENTS AND METHODS: Fourteen patients with erectile failure who had previously responded to intracorporeal injection therapy were enrolled in a randomized placebo-controlled trial. They were given two topical applications, comprising either a cream containing aminophylline, isosorbide dinitrate and co-dergocrine mesylate, or a placebo cream of similar appearance containing no pharmacologically active ingredients. Each patient received 16 applications, eight of the active cream and eight placebo. The creams were applied alternatively on successive occasions and the results recorded. RESULTS: The active cream, applied on 77 occasions, resulted in three good and 13 partial erections. The placebo cream, applied on 76 occasions, yielded four good and 13 partial erections. CONCLUSIONS: We were unable to reproduce the successful results reported by others; in the present study the active cream performed no better than placebo. PMID- 10368203 TI - Intracavernosal versus intraurethral alprostadil: a prospective randomized study. AB - OBJECTIVE: To compare the medicated urethral system for erection (MUSE) with standard intracavernosal prostaglandin E1 (PGE1) in the treatment of erectile dysfunction. PATIENTS AND METHODS: Sixty consecutive men with organic erectile dysfunction were prospectively randomized to receive either 20 microg of intracavernosal PGE1 (group 1, 30 patients) or 1 mg MUSE (group 2, 30 patients). Response to the drugs was recorded in the outpatient clinic and all patients continued a home-treatment programme for 3 months. After each home administration, patients recorded the grade of erection in diaries, whether or not sexual intercourse occurred and any adverse reactions to the drugs. Comfort and ease of administration were also recorded. RESULTS: The characteristics of the patients of both groups were similar; 10 patients in group 1 and 25 in group 2 completed the 3-month treatment programme, i.e. a withdrawal rate of 67% and 17% for groups 1 and 2, respectively (P<0.05). During outpatient dosing, 27 (90%) patients in group 1 and 18 (60%) patients in group 2 achieved a good erection (P<0.05). Intercourse during the 3 months of home treatment was reported at least once in 26 (87%) patients in group 1, compared with 16 (53%) patients in group 2 (P<0.05). After 3 months of home treatment, patients had administered a total of 242 doses of intracavernosal PGE1 and 360 doses of MUSE; intercourse was reported after 206 (85%) and 198 (55%) administrations of PGE1 and MUSE, respectively (P<0.05). The most common adverse reaction was urogenital pain, reported by 14 (47%) patients in group 1 and two (7%) patients in group 2 (P<0.05). Home treatment was assessed as easy by 12 (40%) patients in group 1 and 27 (90%) in group 2 (P<0. 05). CONCLUSION: Although MUSE is less effective than intracavernosal PGE1, it is more attractive and accepted well by most patients as an easy method of treatment with minimal or no discomfort. PMID- 10368204 TI - Repeat epididymovasostomies: are they worthwhile? AB - OBJECTIVE: To establish whether it is worthwhile repeating epididymovasostomy in men with persistent obstructive azoospermia. PATIENTS AND METHODS: The study included 24 men with obstructive azoospermia, persisting after previous surgery for blockage in the body or tail of the epididymes, who underwent repeat epididymovasostomy proximal to the previous anastomoses. Semen was re-analysed after 6 and 12 months, and information about pregnancy self-reported or determined by postal survey. RESULTS: The postoperative sperm concentration was >107 /mL in 15 patients (62%) and 10 female partners became pregnant (41%). Antisperm antibodies were present in nine patients and three of their partners became pregnant after the man received steroid therapy. Unilateral revision did not produce a favourable outcome. CONCLUSION: Having defined a favourable group of men with obstructive azoospermia by scrotal exploration, i.e. those with caudal epididymal blocks and patent vasa deferentia, initial technical failure should not preclude surgical revision of the anastomoses in selected cases. PMID- 10368205 TI - Total urogenital sinus mobilization: expanded applications. AB - OBJECTIVE: To report further applications of total urogenital sinus mobilization, earlier described as an easier method to correct a cloaca. PATIENTS AND METHODS: Seven children (six girls and one boy, mean age 4 years, range 3 months to 10.5 years) underwent surgery and were followed for a mean of 1 year; their diagnoses included persistent cloaca and congenital adrenal hyperplasia (CAH) in two each, and a urogenital sinus (UGS), bladder exstrophy and penile agenesis in one each. The UGS is approached through a posterior sagittal incision and dissected circumferentially to the retropubic space, allowing the UGS to descend. It is then excised and separate openings of the vagina and urethra created. This technique is applicable to a UGS of /=60 years. The frequency of each metabolic abnormality and the value of each urinary constituent were compared among subgroups of age and gender. RESULTS: Hyperoxaluria was the most common abnormality, present in 56% and 67% of patients aged <60 and >/=60 years, respectively. Hyperuricosuria was significantly more common in older than in younger patients. There were no significant differences in the frequencies of hypercalciuria and hypocitraturia between the age groups. The urinary excretion of oxalate and the ratio of oxalate to creatinine were significantly greater in older than in younger men. The frequency of low urine volume was lower in older than in younger patients and the mean urinary volume was also greater in the older group. CONCLUSIONS: Hyperuricosuria and hyperoxaluria seem to be essential risk factors for calcium oxalate stone formation in elderly patients. Urinary oxalate excretion is significantly greater in older than in younger stone formers and is more prominent in men. PMID- 10368231 TI - The role of retroperitoneoscopy in the management of renal and adrenal pathology. AB - OBJECTIVES: To describe the technique, findings and results of retroperitoneoscopic ablation of recalcitrant renal, giant adrenal and complex peripelvic cysts, and nephrectomy for nonfunctioning congenital anomalous kidneys. PATIENTS AND METHODS: Nine patients (six men and three women, mean age 56 years, range 44-68, five with renal, two with adrenal and two with peripelvic cysts, diameter 6-14 cm) were treated by retroperitoneoscopic cyst ablation using three 10-mm ports. Six further patients (two male and four female, mean age 24 years, range 13-38) underwent retroperitoneoscopic nephrectomy using three or four ports for anomalous nonfunctioning kidneys; three patients had a pelvic kidney, two a horseshoe kidney and one an iliac kidney. Isthmusectomy was also performed in the patients with horseshoe kidneys. RESULTS: Retroperitoneoscopic cyst ablation was successful in all nine patients; the mean (range) operative duration was 69 (50-85) min in patients with simple renal and adrenal cysts, and 185 (160-210) min in patients with peripelvic cysts. The mean (range) blood loss was 130 (50-200) mL and hospital stay 2.33 (2-4) days. At the last follow-up, 15 39 months after the procedure, all patients were asymptomatic and satisfied with the outcome, with no recurrence of cysts. Retroperitoneoscopic nephrectomy with isthmusectomy (when applicable) was successful in the six patients with anomalous kidneys, with a mean (range) operative duration of 105 (85-120) min; the mean (range) blood loss was 116 (75-150) mL and the analgesic requirement 208 (150 250) mg of diclofenac sodium. The hospital stay was 2-3 days and the delay before return to preoperative activity 7-14 days. CONCLUSIONS: Retroperitoneoscopic cyst ablation is a safe and effective method to treat symptomatic cysts of the upper urinary tract which are refractory to other forms of management. Dissection is difficult in patients with peripelvic cysts. Retroperitoneoscopic nephrectomy for anomalous kidneys is a challenging procedure because of the abnormal location, anomalous vessels and presence of an isthmus. With advances in laparoscopy and increasing experience, open surgery for such conditions is likely to become obsolete. PMID- 10368232 TI - Papillary renal cell carcinoma: clinicopathological characteristics and evaluation of prognosis in 42 patients. AB - OBJECTIVE: To determine the clinicopathological characteristics of histologically defined papillary renal cell carcinoma (RCC) in relation to prognosis. PATIENTS AND METHODS: In total, 768 patients with RCC underwent nephrectomy at our university hospital between 1957 and 1995. RCC was classified into clear-cell carcinoma in 689 patients (89.7%, no follow-up in 14), chromophobe cell carcinoma in 36 (4.7%, no follow-up in two) and papillary RCC in 43 (5.6%, no follow-up in one). In the present study, the 42 patients with papillary RCC who underwent nephrectomy and were followed up were those in whom the clinicopathological features of the papillary RCC were assessed. Factors assessed were the presence or absence of foam-cell infiltration, occurrence of bleeding and/or necrosis, presence or absence of a pseudocapsule, mixed occurrence with clear-cell carcinoma, presence or absence of solid variants, cytoplasmic appearance (basophilic vs eosinophilic cells), stage, nuclear grade of malignancy, and angiographic appearance in relation to prognosis. The prognosis was also compared among patients with clear-cell, chromophobe cell and papillary RCC. RESULTS: The prognosis was significantly better in patients with foam-cell infiltration (P=0.03), with a pseudocapsule (P=0.07), with no solid variants (P=0.001) and with basophilic cells (P<0.001). There were also significant differences in survival between patients with low-stage (1+2) and high-stage (3+4) disease (P=0.003), and among grades 1-3 (grade 1 vs 2, P=0.05; grade 1 vs 3, P<0.001, grade 2 vs 3, P=0.006). Furthermore, the prognosis in patients with papillary RCC was worse than in those with chromophobe cell carcinoma (P=0.02), but there was no significant difference in survival between patients with papillary RCC and those with clear-cell carcinoma. CONCLUSION: The clinicopathological features (e.g. the presence or absence of foam cells, of a pseudocapsule and of solid variants, cytoplasmic appearance, and the stage and nuclear grade of malignancy) are important prognostic factors for patients with papillary RCC. Furthermore, the prognosis in patients with papillary RCC is similar to those with clear-cell carcinoma. PMID- 10368233 TI - Changes in urinary output during laparoscopic adrenalectomy. AB - OBJECTIVE: To better understand the physiological effects of pneumoperitoneum, by examining changes in urinary output during gaseous and gasless laparoscopic adrenalectomy. PATIENTS AND METHODS: Laparoscopic adrenalectomy was performed with gas in six patients and without in three. Urinary output was measured during insufflation and after desufflation. RESULTS: In all patients who received gas, the urinary output was significantly decreased during insufflation and significantly increased after desufflation. However, there were no changes in urinary output in patients who did not receive gas. CONCLUSION: For the safety of laparoscopic surgery it is important to recognise that oliguria occurs during pneumoperitoneum, although the changes in urinary output caused no complications in renal function. PMID- 10368234 TI - The re-use of irrigating equipment for flexible cystoscopy is not safe. AB - OBJECTIVE: To determine the incidence of fluid reflux from the lower urinary tract into the connecting tubing used for irrigation in patients undergoing flexible cystoscopy. PATIENTS AND METHODS: The study was conducted in 94 consecutive male and female patients attending routine outpatient flexible cystoscopy lists. A sensor was designed and constructed to determine the presence of any retrograde flow of irrigating fluid, and the volume of any reflux through the connecting tubing. The mean (sd) cystoscope internal channel volume was 2.56 (0.25) mL; the level of significant reflux was set at >/=2. 25 mL. RESULTS: Reflux of irrigating fluid occurred in 11 males (17%) and was significant in six (9%) of 65 male patients, with the irrigating fluid reservoir set at a height of 0.78 m above the patient's mid-coronal level. No reflux occurred in the 29 females studied. CONCLUSIONS: Significant reflux can occur in males and hence the connecting tubing should be regarded as contaminated. Infection-control measures must include the prevention of transmission of blood-borne infections, e.g. hepatitis B and C viruses, and human immunodeficiency virus, because of the risk that blood may contaminate urine, and they should be implemented in all cases regardless of patient risk factors. From the available evidence, flexible cystoscopy should always be performed with single-use irrigation systems. PMID- 10368235 TI - Suprapubic catheterization: a suitable procedure for clinical nurse specialists in selected patients. AB - OBJECTIVE: To assess the feasibility and advantages of urology clinical nurse specialists (NSs) extending their professional role to include inserting initial suprapubic catheters in selected patients in the hospital and community setting. PATIENTS AND METHODS: A urology NS, who is also the district continence adviser, was formally taught by a consultant urologist how to insert suprapubic catheters using the 'Add-a-Cath' introducer technique (Femcare, UK). Once deemed competent, the NS was indemnified by the Trust to carry out the procedure in carefully selected patients using a safe and unequivocal protocol. RESULTS: Over a period of 40 months, the NS undertook the procedure in 164 patients, with 64 being catheterized in the community. There were no serious complications in the series and during the period studied, only 17 patients were referred back to the urologists for catheterization under formal theatre conditions. CONCLUSION: Initial suprapubic catheterization can be carried out safely by trained NSs in selected patients. This extended professional role has helped to improve the continuity and quality of care for patients who require long-term catheterization. The NS was also a valuable source of training for junior medical staff who have limited experience with suprapubic catheterization. PMID- 10368236 TI - The long-term outcome in patients with superficial transitional cell carcinoma of the bladder: a single-institutional experience. AB - OBJECTIVE: To determine the natural history of transitional cell carcinoma (TCC) of the bladder, and to identify factors which place patients at lifelong risk of developing progression and dying from bladder carcinoma. PATIENTS AND METHODS: The long-term outcome was evaluated retrospectively in 231 patients with superficial bladder TCC, assessed for the first time within a 6-year period from 1981 to 1986, with a median follow-up of 108 months. Of 231 patients, 217 (94%) were initially treated by transurethral or segmental resection. RESULTS: Recurrence developed in 141 of 217 (65%) patients; the duration of the interval free of recurrence was significantly less for patients with initial G3 tumours than that for those with G1 (P<0.01) and for pT1 compared with pTa disease (P<0.01). The disease progressed in 42 of 231 (18%) patients. Differences in the progression-free interval between patients with G1 and G3 tumours, and with pTa and pT1 disease, were statistically significant (P<0. 005 and P<0.001, respectively). In 27 of 231 patients (12%), TCC of the bladder was the cause of death, whilst 118 (51%) died from unrelated causes. There were no deaths among patients with initial pTaG1 tumours, compared with 10 of 26 (38%) deaths in those with pT1G3 disease at presentation. CONCLUSION: The long-term prognosis is good for patients with pTaG1 tumours, whilst pT1G3 is a potentially aggressive disease. Lifelong endoscopic surveillance is mandatory in patients in whom new tumours are very active, at least in those of younger age. Routine cystoscopy can possibly be discontinued in patients with low-grade, low-stage disease in whom a low liability of recurrence has been shown during follow-up. PMID- 10368237 TI - Radical cystoprostatectomy combined with Mainz pouch bladder substitution to the urethra: long-term results. AB - OBJECTIVE: To analyse, in a retrospective study, the oncological outcome, pouch related complications, continence and micturition after radical cystoprostatectomy combined with Mainz pouch orthotopic bladder substitution to the urethra for the treatment of bladder cancer. PATIENTS AND METHODS: Between 1986 and 1996, three urological departments contributed 108 male patients to the review. The same exclusion criteria from orthotopic bladder substitution were applied by all centres, i.e. multifocal or concomitant carcinoma in situ, tumour at the bladder neck, positive biopsy from the prostatic urethra, locally advanced tumour and lymph node involvement. In all, 103 patients were evaluable for follow up, with a mean (range) follow-up of 42 (3-132) months. RESULTS: Pathological examination of the cystectomy specimen revealed 81% organ-confined tumours. During follow-up, 84% of patients remained free of tumour, 7% developed distant metastases, 5% local recurrences, 4% urethral recurrences, and 1% upper tract urothelial cancer; 85% of patients are capable of spontaneous voiding, with a mean pouch capacity of 720 mL. Daytime continence was achieved in 88%, including 17% wearing one safety pad; 9% had stress incontinence and 3% total incontinence; 67% could sleep through the night, with either complete continence (34%) or one safety pad (33%). Nocturnal incontinence occurred in 11%. Uretero-intestinal stenosis occurred in 15 of 205 (7%) renal units, requiring ureteric reimplantations in 11, nephrectomy in three and antegrade dilatation in one. Reflux was not noted in any patient. About half the patients were on anti acidotic prophylaxis. CONCLUSION: The large bowel segment in the Mainz-pouch technique of orthotopic bladder substitution provides good reservoir capacity and continence rates, with less ileum used than in all-ileum pouches. The surgical technique is simple and reproducible, and in particular the antireflux ureteric implantation into the caecum protects the upper urinary tracts. PMID- 10368238 TI - Urinary continence and erectile function after bladder neck sling suspension in male patients with spinal dysraphism. AB - OBJECTIVE: To assess the outcome of using sling suspensions combined with clean intermittent catheterization (CIC) in patients with spina bifida, of whom a third are incontinent through pelvic floor paralysis. PATIENTS AND METHODS: Between March 1992 and April 1997, 14 male patients (mean age at surgery 11.7 years, range 6.5-15.2) with spina bifida and neurogenic sphincter incontinence underwent a puboprostatic sling suspension as a primary treatment. The procedure, via an abdominoperineal approach, consists of suspending the bladder neck by placing a simple U-shaped rectus abdominus fascial sling. The perineal approach is used to develop the plane between the rectum and Denonvillier's fascia, and to prepare the passage of the sling alongside the prostate. Apart from the sling procedure, eight of the 14 patients underwent autoaugmentation of the bladder and two underwent ileocystoplasty during the same operation. All patients used CIC daily. Erectile function was assessed by reports from the patients and their parents, and continence by report and urodynamic studies. RESULTS: Of the 14 patients, 13 achieved urinary continence with no additional procedures; one required a subsequent submucosal injection at the suspension site with silicone particles in povidone (Macroplastique(R)) to become continent. Two patients reported slight leakage at night. Before surgery, all but one patient reported having spontaneous or mechanically manipulated erections; none had erections on psychological stimulation. After surgery, erectile function was preserved in 13 of the 14 patients; in one there were problems establishing the right dissection plane between the rectum and prostate, but spontaneous erections returned a year after surgery. CONCLUSION: In males, the abdominoperineal puboprostatic sling suspension using rectus abdominis fascia appears to be a successful treatment for sphincter incontinence in patients with spina bifida, and safely maintains erectile function. PMID- 10368239 TI - beta-sitosterol for the treatment of benign prostatic hyperplasia: a systematic review. AB - OBJECTIVES: To conduct a systematic review of the evidence for the efficacy of beta-sitosterol in men with symptomatic benign prostatic hyperplasia (BPH). METHODS: Studies were identified through Medlinetrade mark (1966-98), EMBASEtrade mark, Phytodok, the Cochrane Library, bibliographies of identified trials and review articles, and contact with study authors and pharmaceutical companies. Randomized trials were included if: men had symptomatic BPH; plant extract preparations contained beta-sitosterols; a control group received placebo or a pharmacological therapy; and treatment duration was >/=30 days. Study characteristics, demographic information, enrolment criteria and outcomes were extracted. RESULTS: Four trials comprising a total of 519 men met the inclusion criteria. All were double-blind and lasted 4-26 weeks. Three studies used nonglucosidic beta-sitosterols and one used a preparation that contained only beta-sitosterol-beta-d-glucoside. Compared with placebo, beta-sitosterol improved urinary symptom scores and flow measures. For the two studies reporting the International Prostate Symptom Score (IPSS), the weighted mean difference (WMD) against placebo was -4.9 IPSS points (95% confidence interval, CI,-6.3 to-3.5). The WMD for peak urinary flow rate was 3.91 mL/s (95% CI 0.91 to 6.90, four studies) and for residual volume the WMD was -28.62 mL (95% CI-41.42 to-15.83, four studies). beta-sitosterol did not reduce prostate size. The trial using pure beta-sitosterol-beta-d-glucoside (WA184) showed no improvement in urinary flow measures. Withdrawal rates for men assigned to beta-sitosterol and placebo were 7.8% and 8.0% (not significant), respectively. CONCLUSION: beta-sitosterol improves urological symptoms and flow measures. However, the existing studies are limited by short treatment duration and lack of standardized beta-sitosterol preparations. Their long-term effectiveness, safety and ability to prevent the complications of BPH are unknown. PMID- 10368240 TI - Thrombotic risk factors associated with transurethral prostatectomy. AB - OBJECTIVE: To ascertain the potential thrombotic risk associated with transurethral prostatectomy (TURP). PATIENTS AND METHODS: The changes in coagulation variables were assessed in a prospective study of 40 patients undergoing TURP. RESULTS: There was a significant increase in thrombin antithrombin complexes 6 h after TURP (anova, P=0.01) combined with a significant decrease in activated partial thromboplastin time (anova, P=0.006), suggesting a postoperative hypercoagulable state. The significant increase in d-dimer 24 h after TURP (anova, P=0.015) in the absence of any significant rise in tissue plasminogen activator antigen levels perioperatively (anova, P=0.737) suggests a physiological fibrinolytic response to the developing procoagulant state. The absence of any significant increase in plasminogen activator inhibitor-1 antigen perioperatively (anova, P=0.348) suggests the observed hypercoagulability is not due to a 'fibrinolytic shutdown' reported in other forms of surgery. CONCLUSION: TURP is associated with a hypercoagulable prothrombotic state; aspirin withdrawal perioperatively may be hazardous, and low-dose heparin prophylaxis for venous thrombosis should be considered. PMID- 10368241 TI - Bleeding and activation of coagulation during and after transurethral prostatectomy: importance of the acute-phase response and prostate specific antigen? AB - OBJECTIVE: To evaluate the importance of coagulation activation in patients with benign prostatic hyperplasia, undergoing transurethral prostatic resection (TURP) and to examine whether changes in activity are related to blood loss, the circulatory entry of prostate specific antigen (PSA), operative trauma (resected tissue weight) and the inflammatory response, as assessed by C-reactive protein (CRP). PATIENTS AND METHODS: TURP was performed in 24 men and the weight of resected tissue and blood loss determined. The activation of coagulation was followed using new sensitive and specific assays, and the changes related to blood loss, the release of PSA, operative trauma and the acute-phase response. The area under the curve (AUC) for the measured quantities was used in correlation analysis. RESULTS: TURP was followed by a marked activation in coagulation. There was no correlation between the markers of coagulation and the operative blood loss, but the latter correlated with the weight of resected tissue (P=0.001). Postoperatively, the blood loss correlated with prothrombin fragment (F1+2; P=0.010), with thrombin-antithrombin complexes (TAT; P=0.024), and with the PSA concentrations (P=0.016) but not with fibrinogen. Serum concentrations of PSA increased significantly and the AUC in the operative period correlated with F1+2 (P=0.003) and TAT (P<0. 005), but postoperatively only with F1+2 (P=0.013). The weight of resected tissue correlated operatively with PSA (P=0.012) but not with the concentrations of F1+2 or TAT. Postoperatively, there was a correlation with the acute-phase proteins, CRP (P=0.005), fibrinogen (P=0.012) and with PSA (P=0.020). CONCLUSION: The operative blood loss is caused by surgical factors and the observed postoperative hypercoagulable state can be explained as a physiological response to bleeding, i.e. to secure haemostasis. The activity of coagulation was unrelated to operative trauma, but the acute phase proteins were. The release of PSA into the circulation probably has an effect on blood coagulation. PMID- 10368242 TI - Serum insulin-like growth factor-1 is not a useful marker of prostate cancer. AB - OBJECTIVE: To determine whether the use of serum insulin-like growth factor 1 (IGF-1) levels is more efficient than serum prostate specific antigen (PSA) levels in predicting prostate cancer in patients undergoing prostatic biopsy. PATIENTS AND METHODS: The study included 94 consecutive patients who required transrectal ultrasonography (TRUS)-guided biopsies of their prostate and who had blood samples taken before their biopsies. These samples were then analysed for IGF-1 and PSA concentrations. Six prostatic biopsies were taken from each patient; they were assessed and a diagnosis made of prostate cancer or no malignancy. RESULTS: Thirty-seven patients were found to have prostate cancer and 57 had no evidence of malignancy. There was no statistical difference in serum IGF-1 levels between these groups. The PSA level and age of the patients differed significantly between the groups (both P<0.001). There was no correlation between IGF-1 and PSA levels, and even when the age difference in the groups was considered, there was still no significant relationship between IGF-1 levels and the incidence of prostate cancer. In patients with a PSA level of 4-20 microg/L there was no statistically significant difference in IGF-1 levels between the groups. CONCLUSION: Serum IGF-1 as a tumour marker does not help to predict patients with prostate cancer. PSA level and even age were better predictors of the presence of prostate cancer than were serum IGF-1 levels. PMID- 10368243 TI - Free prostate-specific antigen 'in the field': a useful adjunct to standard clinical practice. AB - OBJECTIVES: To determine the clinical utility of the ratio of free to total prostate-specific antigen (f/tPSA), which may improve the discrimination of PSA testing in the assessment of patients with prostatic disease, in a busy clinical practice. PATIENTS AND METHODS: A series of 198 men undergoing transrectal ultrasonography (TRUS)-guided biopsy, because of a high total PSA level or an abnormal digital rectal examination or both, had blood samples taken for the assessment of both free and total PSA before any form of prostatic manipulation. The histological findings were compared with tPSA, fPSA and f/tPSA, evaluating three different thresholds of f/tPSA. RESULTS: PSA levels, PSA density and fPSA density differed significantly between those with benign histology and neoplasia. The f/tPSA only differed significantly in men with a PSA level of 10. 1-15 ng/mL. The f/tPSA threshold of 0.25 was the most useful clinically, with a negative predictive value for prostate cancer of 91%. Using this threshold would reduce the negative biopsy rate by 30%. CONCLUSION: The f/tPSA is of clinical value in reducing the negative biopsy rate and in the management of patients with a high PSA level and previous negative biopsies. PMID- 10368244 TI - The free-to-total serum prostatic specific antigen ratio as a predictor of the pathological features of prostate cancer. AB - OBJECTIVES: To analyse the role of the ratio of serum free (f) to total (t) prostatic specific antigen (f/tPSA) as a predictor of the pathological features in patients with clinical stage T1c disease submitted for radical prostatectomy. PATIENTS AND METHODS: Total and fPSA were determined before surgery in 76 consecutive patients with clinical stage T1cN0M0 disease and a serum tPSA level of 4-20 ng/mL. The pathological stage was defined as organ-confined in 47 (62%), with capsular penetration in 27 (35%) and seminal vesicle infiltration in two (3%). In 18 of the specimens (24%) the surgical margins were positive. The pathology was favourable in 41 patients (55%). Total and fPSA were determined using the Tandem and Tandem-free PSA immunoassays (Hybritech Inc, Belgium) RESULTS: The mean tPSA was: 8.7 ng/mL when the tumour was organ-confined and 8.6 ng/mL when the disease was extraprostatic (P>0.05); 8.4 ng/mL when the tumour was specimen-confined and 9.3 ng/mL when positive margins or seminal vesicle involvement were detected (P>0.05); and 8.3 ng/mL when the pathology was favourable and 9.0 ng/mL when unfavourable (P>0.05). The f/tPSA was 11.8% when the tumour was organ-confined and 9.1% when it was not (P<0.03), 11.9% when the tumour was specimen-confined and 7.6% when not (P<0.002) and 12.1% when the pathology was favourable and 9.1% when unfavourable (P<0.008). A threshold of 11% f/tPSA provided an odds ratio for organ-confined disease of 2.7, for specimen confined disease of 7.6 and for a favourable pathology of 3.9. CONCLUSION: The f/tPSA seems to provide useful information in the prediction of the pathological features of patients with clinical T1c prostate cancer and a tPSA of 4.1-20 ng/mL. PMID- 10368245 TI - The rectal administration of lidocaine gel and tolerance of transrectal ultrasonography-guided biopsy of the prostate: a prospective randomized placebo controlled study. AB - OBJECTIVE: To evaluate the effect of the rectal administration of lidocaine gel on the tolerance of systematic sextant transrectal ultrasonography (TRUS)-guided prostatic biopsies. PATIENTS AND METHODS: From January to September 1997, patients undergoing initial biopsy mapping of the prostate (with six systematic TRUS-guided transrectal biopsies) were randomized using a pre-established randomization list into two groups. In group 1, 15 mL of 2% lidocaine gel (Astra, Sodertalje, Sweden) was administered intrarectally 15 min before the biopsies. In group 2 (placebo), 15 mL of trans-sonic hydrophilic gel (Rivadis Laboratory, Thouars, France) was administered transrectally under the same conditions. Patients were randomized and the gel administered by a nurse; neither the patients nor the urologists were aware of which product was administered. At the end of the procedure, patients were asked to score the severity of discomfort of the biopsies, using a self-administered rating scale. RESULTS: In all, 109 patients were included, in either group 1 (56 patients) or group 2 (53 patients). Slight pain or no pain was experienced by the vast majority of patients in both groups. Moderate to severe pain was experienced in 12.5% of patients in group 1 and 11.3% of patients in group 2. There was no difference in patient tolerance between the groups (P=0.39). Only minor complications occurred and complication rates were not significantly different between the groups. CONCLUSION: The rectal administration of lidocaine has no impact on the tolerance to prostatic biopsy. PMID- 10368247 TI - Biofeedback or pelvic floor muscle exercises for female genuine stress incontinence: a meta-analysis of trials identified in a systematic review. AB - OBJECTIVE: To test, by meta-analysis, the conclusion of a systematic review that biofeedback was no more effective than pelvic floor muscle exercises alone for the treatment of female genuine stress urinary incontinence. MATERIALS AND METHODS: Data extracted from the five trials identified in the systematic review were subjected to pooled analysis of odds ratios for the outcome of 'cure'. RESULTS: The odds ratio for biofeedback combined with pelvic floor muscle exercises, compared with pelvic floor muscle exercises alone, leading to cure was 2.1 (95% confidence interval 0.99-4.4). CONCLUSIONS: Biofeedback may be an important adjunct to pelvic floor muscle exercises alone in the treatment of female genuine stress urinary incontinence. A quantitative statistical analysis of the studies identified leads to different conclusions from those in the systematic review. PMID- 10368246 TI - The pubofascial anchor sling procedure for recurrent genuine urinary stress incontinence. AB - OBJECTIVE: To evaluate, in a preliminary study, the outcome of a modified pubovaginal sling operation with titanium bone anchors for recurrent genuine stress urinary incontinence (GSI) in women. PATIENTS AND METHODS: This prospective study included 13 consecutive women who underwent the modified sling procedure between September 1994 and August 1996. The subjective and objective cure of urinary stress incontinence, and the occurrence of postoperative osteitis pubis, were assessed. RESULTS: All 13 patients subjectively claimed complete urinary continence and 12 were objectively cured (12 patients agreed to undergo a repeat urodynamic study) during a median (range) follow-up of 26 (19-38) months. There were no cases of postoperative osteitis pubis, bladder injury or major complications. Mild suprapubic pain was a frequent and self-limiting complication. CONCLUSIONS: This innovative modified sling procedure is effective for recurrent urinary stress incontinence, with no complication of osteitis pubis. We suggest that this procedure should be considered as a treatment for recurrent GSI and perhaps for primary GSI. A study incorporating a longer follow up and more patients has been planned. PMID- 10368248 TI - Nocturia and polyuria in men referred with lower urinary tract symptoms, assessed using a 7-day frequency-volume chart. AB - OBJECTIVES: To investigate if a 7-day frequency-volume (FV) chart could identify nocturia on a polyuric basis in patients with lower urinary tract symptoms (LUTS) suggestive of benign prostatic hyperplasia (BPH). PATIENTS AND METHODS: The study included 23 patients (mean age 62.8 years, range 42-78) with LUTS who were referred for the evaluation of potential BPH and 11 men (control subjects, mean age 63.3 years, range 58-69); all completed a 7-day FV chart investigation as outpatients. RESULTS: Nocturia was associated with nocturnal polyuria in 10 of 23 patients with LUTS; these 10 patients had a diminished diurnal variation of urine production, whereas 13 patients had a diurnal variation in urine production comparable with that in controls with no nocturia. The degree of nocturia correlated positively with nocturnal urine production but showed no relationship with sleep duration. The nocturnal polyuria in these patients was associated with a higher 24-h urine production and seemed at least partly to be caused by a higher fluid intake during daytime. CONCLUSION: Nocturia on a polyuric basis can be detected by using a FV chart. In these patients, a 3-day FV chart would be sufficient to detect nocturia on a polyuric basis and seems therefore to be a valuable tool in evaluating patients with LUTS referred for potential BPH. PMID- 10368249 TI - Intra-uterine testicular torsion: early diagnosis and treatment. AB - OBJECTIVE: To review the incidence and treatment of intra-uterine torsion of the testis which although rare is being recognized with increasing frequency. PATIENTS AND METHODS: From 1988 to 1997, five newborns (mean birth weight 3.62 kg, range 3.15-4.12) with unilateral torsion of the testis were treated; all underwent emergency exploration. The right testis was affected in three and the left in two boys. RESULTS: In all except one child, the affected testis was enlarged, firm to hard, tender, the overlying skin dark red and the affected testis higher than the contralateral testis. In one child the right testis was enlarged and higher, but soft to firm, and the overlying skin was oedematous and red. The exploration revealed extravaginal torsion of the testis which was gangrenous in four; in one after detorsion there was haemorrhage and haematoma of the cord and the tunica, and the testis was slightly congested but not gangrenous. This testis was preserved and bilateral orchidopexies performed; at 18 months both testes are palpable and of normal size. In the remaining four children the testes were frankly necrotic; they underwent orchidectomy and contralateral orchidopexy. Histology in all four revealed a totally infarcted testis with extensive haemorrhage and vascular congestion. CONCLUSION: The early diagnosis and treatment of intra-uterine torsion of the testis is essential. PMID- 10368250 TI - The outcome of artificial urinary sphincter placement after a mean 15-year follow up in a paediatric population. AB - OBJECTIVE: To evaluate the long-term outcome in children who had an artificial urinary sphincter (AUS) placed, after a minimum of 10 years of follow-up. PATIENTS AND METHODS: The medical records of patients who had an AUS placed at the Children's Hospital of Michigan were reviewed and a telephone questionnaire was then completed by all patients with an AUS currently in place. RESULTS: Forty seven children initially had an AUS placed between October 1978 and August 1986; medical records and follow-up were available for 32. After a mean follow-up of 15.4 years, 13 patients had had the AUS removed and 19 currently have an intact AUS. Erosion or infection was responsible for all AUS removals. Possible risk factors for AUS removal were prior AUS erosion, prior bladder neck surgery and a balloon pressure of >70 cmH2O. Eighteen of 19 patients with an intact AUS are dry and seven void volitionally. Revision was the most common reason for additional surgery, but the revision rate has decreased with the most current AS-800 model, to 0.03 revisions per patient-year. Of the 13 patients with an AS-800 model placed after 1987, nine have not required revision. Upper tract changes were mild and uncommon. CONCLUSION: The AUS is a durable and effective surgical option in the management of neurogenic urinary incontinence, and is the only reliable technique that can preserve volitional voiding. With technical improvements to the AUS and a longer follow-up, the revision rate has decreased. Causes of AUS removal may be preventable with improvements in surgical technique and patient selection. AUS placement should be considered as a first choice for the surgical management of neurogenic sphincteric incompetence. PMID- 10368251 TI - Hypospadias repair by skin flaps: a comparison of onlay preputial island flaps with either Mathieu's meatal-based or Duckett's tubularized preputial flaps. AB - OBJECTIVES: To evaluate the success of different skin flaps in the one-stage correction of primary hypospadias, with particular emphasis on comparing onlay preputial island flaps with Mathieu's meatal-based and Duckett's preputial tubularized flaps. PATIENTS AND METHODS: During a 12-year period, 418 patients underwent single-stage primary hypospadias repair using skin flaps, carried out by one surgeon. The surgical techniques used included Mathieu's repair in 216 (52%), Duckett's in 148 (35%), onlay preputial flaps in 42 (10%) and the Mustarde flap procedure in 12 (3%). The surgical results were reviewed, assessing complications and the functional and cosmetic outcome. RESULTS: At a mean follow up of 23 months the initial overall complication rate for flap procedures was 22%; however, after a mean of 1.4 procedures, the final success rate was 95%. The complication rate was significantly (P<0.05) higher in patients with a proximal urethral meatus, with severe chordee or in repairs involving transection of the urethral plate. However, the complication rates were not significantly different when the patients underwent repair when aged <2 years or >2 years. Despite no significant difference in overall complication rates, onlay procedures tended to be used in more severe hypospadias than was Mathieu's repair. Duckett's repair caused a significantly higher overall complication rate as fistulae, strictures, meatal stenoses and tubular abnormalities than did onlay procedures. The use of double-faced preputial island flaps resulted in an inferior cosmetic appearance than the use of single-faced flaps, but the overall complication rates did not differ significantly between these techniques. CONCLUSIONS: Hypospadias repair using skin flaps offered a reliable and durable outcome. However, complication rates were greater in patients with severe hypospadias and with techniques requiring transection of the urethral plate. The onlay preputial island-flap technique was more widely applicable than was Mathieu's repair and had a lower complication rate than Duckett's procedure. PMID- 10368252 TI - The expression of PAX5 in human transitional cell carcinoma of the bladder: relationship with de-differentiation. AB - OBJECTIVE: To investigate the expression of PAX genes, a family of developmental control genes (which encode nine nuclear transcription factors essential for embryogenesis and are proto-oncogenes in mice) in human transitional cell carcinoma (TCC) of the bladder. MATERIALS AND METHODS: PAX gene expression was assessed in three established bladder cancer cell lines and 29 primary tumours using the reverse transcriptase-polymerase chain reaction and Southern analysis. RESULTS: All three established TCC cell lines and 79% of primary TCCs expressed PAX5 mRNA. There was a significantly higher proportion of PAX5 expression in malignant than in benign urothelium (P=0.02, Fisher's exact test); nine of 12 pTa tumours (mucosa-confined), seven of eight pT1 (invading lamina propria) and eight of nine pT2 (invading muscle) expressed PAX5. A higher proportion of tumours with increasing de-differentiation expressed PAX5, which correlates well with the expression pattern of PAX5 in development. In well-differentiated tumours (grade 1), half expressed PAX5, compared with 84% of moderately to poorly differentiated tumours (grades 2/3). The odds ratio for PAX5 expression in malignancy suggests that it increases the risk of malignancy four-fold. CONCLUSION: These data support a role for the PAX family in oncogenesis, by identifying another human neoplasm in which they are inappropriately expressed. PAX5 expression in undifferentiated TCC cells may contribute to pathogenesis by supporting cellular proliferation in the de-differentiated state. Furthermore, the high incidence of PAX5 expression suggests its potential use as a diagnostic tool and therapeutic target in TCC. PMID- 10368253 TI - Immunohistochemical detection of neuronal plexuses and nerve cells within the upper urinary tract of pigs. AB - OBJECTIVE: To investigate the distribution and topography of nervous structures within the renal pelvis and upper part of the ureter of pigs, and thus help to determine the origin, propagation and mechanisms of the modulation of pelvi ureteric peristalsis. MATERIALS AND METHODS: Whole-mount preparations of renal pelves from adult pigs were stained using a universal immunostaining method with streptavidin-alkaline phosphatase. Anti-neuron-specific enolase antibody and anti neurofilament antibody were used as neuronal markers. RESULTS: The patterns of neuronal structures differed between the renal calyces, renal pelvis and upper ureter. In the calyx, there was one single dense nerve plexus; this network contained relatively thin nerve fibres running both circularly and longitudinally. In the wall of the renal pelvis and upper ureter there were two neuronal plexuses, one submucosal and another within the muscular layer; these nerve fibres were mainly orientated longitudinally. Some single nerve cells were also found at the pelvicalyceal border. CONCLUSIONS: These findings suggest a potent nervous system within the upper urinary tract of pigs that connects the renal calyces with the renal pelvis, pelvi-ureteric junction and ureter. The presence of these dense neuronal networks and nerve cells within the wall of the renal pelvis and ureter suggests that propagation, coordination and modulation of pelvi-ureteric peristalsis in pigs may arise through intrinsic nervous stimulation. PMID- 10368254 TI - In vitro investigation of the bladder-vascular selectivity of levcromakalim and YM934 in human tissues. AB - OBJECTIVE: To examine the potencies of the potassium-channel openers (KCOs) levcromakalim and YM934 as relaxing agents on human detrusor and human mesenteric artery smooth muscle, and to determine their bladder-vascular selectivity in vitro. MATERIALS AND METHODS: Strips of human detrusor muscle and mesenteric artery (with the endothelium removed) were set up in physiological salt solution and the tension developed by the tissues recorded. Tissues were precontracted with a concentration of carbachol (detrusor) or phenylephrine (artery) which caused 80% of maximal contraction, and relaxation responses to levcromakalim and YM934 were then obtained. RESULTS: Both KCOs caused relaxation of the bladder detrusor muscle and the mesenteric artery. Maximal responses, when plotted as a percentage of the precontraction, were greater for both KCOs in the bladder muscle than the artery, but the differences were small and not statistically significant. The sensitivity (drug potency) of the detrusor muscle to the KCOs was more than twice that of the artery but this selectivity was only statistically significant for YM934. CONCLUSIONS: Only minor bladder-vascular selectivity for levcromakalim and YM934 could be detected in vitro. This suggests that neither drug would be tolerated clinically, although the results suggest that further development of bladder-selective KCO agents appears feasible. PMID- 10368255 TI - The role of vitamin A in preventing renal scarring secondary to pyelonephritis. AB - OBJECTIVE: To evaluate the efficiency of exogenously administered vitamin A in preventing renal scarring caused by experimental pyelonephritis in rats. MATERIALS AND METHODS: Forty Wistar rats were injected with 0.1 mL of solution containing Escherichia coli (1010 /mL) into both renal medullae. Five equal groups were then formed: group 1 was treated only with ciprofloxacin (30 mg/kg per day, twice daily, intraperitoneally) for 5 days, starting 3 days after bacterial inoculation; in group 2, 60 kIU of vitamin A was injected intramuscularly with the bacterial inoculation; in group 3, 60 kIU of vitamin A was injected similarly, but 3 days after bacterial inoculation; in group 4, 60 kIU of vitamin A was given orally with the bacterial inoculation; and group 5 was treated with ciprofloxacin for 5 days and vitamin A intramuscularly from the third day after bacterial inoculation. All the rats were killed 6 weeks after bacterial injection; blood samples were obtained to determine serum vitamin A and beta-carotene levels, and both kidneys were examined pathologically for scarring, graded as 0 (none), 1 (mild), 2 (moderate) and 3 (severe). RESULTS: Serum vitamin A levels were higher in the rats given vitamin A (group 2-5) than in group 1, being highest in group 4, although only this group had significantly higher levels of vitamin A than group 1 (P<0.05). Histopathologically, the fibrosis was mildest in groups 2 and 4 (two of 16 kidneys grade 1), whereas it was most severe in group 1 (all 16 grade 2-3). Fibrosis was significantly less in groups 2-5 than in group 1 (P<0.05). There was a significant negative correlation between vitamin A levels and the sum of the fibrosis, inflammation and tubular atrophy scores of all rats (r=-0.391, P<0.02). beta-carotene levels were unrelated to renal scarring. CONCLUSION: The administration of vitamin A could have a role in preventing renal scar formation from pyelonephritis induced experimentally in rats. PMID- 10368257 TI - Point of technique: An S-shaped guide tube to facilitate the passage of a JJ ureteric stent at flexible cystoscopy in female patients. PMID- 10368256 TI - Point of technique. The tunnelled suprapubic catheter. PMID- 10368258 TI - Hydatid renal abscess: a report of two cases. PMID- 10368259 TI - Duodeno-ureteric fistula. PMID- 10368260 TI - Neonatal testicular loss secondary to perinatal trauma in breech presentation. PMID- 10368261 TI - Penile entrapment in a bottle: the case for using a diamond-tipped portable glass saw. PMID- 10368262 TI - A urethral duplication cyst complicated by a squamous cell carcinoma. PMID- 10368263 TI - An unusual case of renal vein thrombosis in a man taking sumatriptan. PMID- 10368264 TI - Preputial calculi. PMID- 10368265 TI - Kidney metastasis of lung cancer presenting as renal carbuncle. PMID- 10368266 TI - Acute urinary retention: an unusual presentation of a spinal arteriovenous malformation. PMID- 10368267 TI - Endoscopic incision for the treatment of a ureteric valve. PMID- 10368268 TI - Solution conformation of a parallel DNA triple helix with 5' and 3' triplex duplex junctions. AB - BACKGROUND: Polypurine x polypyrimidine sequences of DNA can form parallel triple helices via Hoogsteen hydrogen bonds with a third DNA strand that is complementary to the purine strand. The triplex prevents transcription and could therefore potentially be used to regulate specific genes. The determination of the structures of triplex-duplex junctions can help us to understand the structural basis of specificity, and aid in the design of optimal antigene oligonucleotides. RESULTS: The solution structures of the junction triplexes d(GAGAGACGTA)-X-(TACGTCTCTC)-X-(CTCTCT) and d(CTCTCT)-X-(TCTCTCAGTC)-X (GACTGAGAGA) (where X is bis(octylphosphate) and nucleotides in the triplex regions are underlined) have been solved using nuclear magnetic resonance (NMR) spectroscopy. The structure is characterised by significant changes in the conformation of the purine residues, asymmetry of the 5' and 3' junctions, and variations in groove widths associated with the positive charge of the protonated cytosine residues in the third strand. The thermodynamic stability of triplexes with either a 5' or a 3'CH+ is higher than those with a terminal thymidine. CONCLUSIONS: The observed sequence dependence of the triplex structure, and the distortions of the DNA at the 5' and 3' termini has implications for the design of optimal triplex-forming sequences, both in terms of the terminal bases and the importance of including positive charges in the third strand. Thus, triplex stabilising ligands might be designed that can discriminate between TA x T-rich and CG x C+-rich sequences that depend not only on charge, but also on local groove widths. This could improve the stabilisation and specificity of antigene triplex formation. PMID- 10368269 TI - Desulfovibrio desulfuricans iron hydrogenase: the structure shows unusual coordination to an active site Fe binuclear center. AB - BACKGROUND: Many microorganisms have the ability to either oxidize molecular hydrogen to generate reducing power or to produce hydrogen in order to remove low potential electrons. These reactions are catalyzed by two unrelated enzymes: the Ni-Fe hydrogenases and the Fe-only hydrogenases. RESULTS: We report here the structure of the heterodimeric Fe-only hydrogenase from Desulfovibrio desulfuricans - the first for this class of enzymes. With the exception of a ferredoxin-like domain, the structure represents a novel protein fold. The so called H cluster of the enzyme is composed of a typical [4Fe-4S] cubane bridged to a binuclear active site Fe center containing putative CO and CN ligands and one bridging 1, 3-propanedithiol molecule. The conformation of the subunits can be explained by the evolutionary changes that have transformed monomeric cytoplasmic enzymes into dimeric periplasmic enzymes. Plausible electron- and proton-transfer pathways and a putative channel for the access of hydrogen to the active site have been identified. CONCLUSIONS: The unrelated active sites of Ni Fe and Fe-only hydrogenases have several common features: coordination of diatomic ligands to an Fe ion; a vacant coordination site on one of the metal ions representing a possible substrate-binding site; a thiolate-bridged binuclear center; and plausible proton- and electron-transfer pathways and substrate channels. The diatomic coordination to Fe ions makes them low spin and favors low redox states, which may be required for catalysis. Complex electron paramagnetic resonance signals typical of Fe-only hydrogenases arise from magnetic interactions between the [4Fe-4S] cluster and the active site binuclear center. The paucity of protein ligands to this center suggests that it was imported from the inorganic world as an already functional unit. PMID- 10368270 TI - An archetypical extradiol-cleaving catecholic dioxygenase: the crystal structure of catechol 2,3-dioxygenase (metapyrocatechase) from Ppseudomonas putida mt-2. AB - BACKGROUND: Catechol dioxygenases catalyze the ring cleavage of catechol and its derivatives in either an intradiol or extradiol manner. These enzymes have a key role in the degradation of aromatic molecules in the environment by soil bacteria. Catechol 2, 3-dioxygenase catalyzes the incorporation of dioxygen into catechol and the extradiol ring cleavage to form 2-hydroxymuconate semialdehyde. Catechol 2,3-dioxygenase (metapyrocatechase, MPC) from Pseudomonas putida mt-2 was the first extradiol dioxygenase to be obtained in a pure form and has been studied extensively. The lack of an MPC structure has hampered the understanding of the general mechanism of extradiol dioxygenases. RESULTS: The three dimensional structure of MPC has been determined at 2.8 A resolution by the multiple isomorphous replacement method. The enzyme is a homotetramer with each subunit folded into two similar domains. The structure of the MPC subunit resembles that of 2,3-dihydroxybiphenyl 1,2-dioxygenase, although there is low amino acid sequence identity between these enzymes. The active-site structure reveals a distorted tetrahedral Fe(II) site with three endogenous ligands (His153, His214 and Glu265), and an additional molecule that is most probably acetone. CONCLUSIONS: The present structure of MPC, combined with those of two 2,3-dihydroxybiphenyl 1,2-dioxygenases, reveals a conserved core region of the active site comprising three Fe(II) ligands (His153, His214 and Glu265), one tyrosine (Tyr255) and two histidine (His199 and His246) residues. The results suggest that extradiol dioxygenases employ a common mechanism to recognize the catechol ring moiety of various substrates and to activate dioxygen. One of the conserved histidine residues (His199) seems to have important roles in the catalytic cycle. PMID- 10368271 TI - Structure of the adenylation domain of an NAD+-dependent DNA ligase. AB - BACKGROUND: DNA ligases catalyse phosphodiester bond formation between adjacent bases in nicked DNA, thereby sealing the nick. A key step in the catalytic mechanism is the formation of an adenylated DNA intermediate. The adenyl group is derived from either ATP (in eucaryotes and archaea) or NAD+4 (in bacteria). This difference in cofactor specificity suggests that DNA ligase may be a useful antibiotic target. RESULTS: The crystal structure of the adenylation domain of the NAD+-dependent DNA ligase from Bacillus stearothermophilus has been determined at 2.8 A resolution. Despite a complete lack of detectable sequence similarity, the fold of the central core of this domain shares homology with the equivalent region of ATP-dependent DNA ligases, providing strong evidence for the location of the NAD+-binding site. CONCLUSIONS: Comparison of the structure of the NAD+4-dependent DNA ligase with that of ATP-dependent ligases and mRNA capping enzymes demonstrates the manifold utilisation of a conserved nucleotidyltransferase domain within this family of enzymes. Whilst this conserved core domain retains a common mode of nucleotide binding and activation, it is the additional domains at the N terminus and/or the C terminus that provide the alternative specificities and functionalities in the different members of this enzyme superfamily. PMID- 10368272 TI - The crystal structure of plasminogen activator inhibitor 2 at 2.0 A resolution: implications for serpin function. AB - BACKGROUND: Plasminogen activator inhibitor 2 (PAI-2) is a member of the serpin family of protease inhibitors that function via a dramatic structural change from a native, stressed state to a relaxed form. This transition is mediated by a segment of the serpin termed the reactive centre loop (RCL); the RCL is cleaved on interaction with the protease and becomes inserted into betasheet A of the serpin. Major questions remain as to what factors facilitate this transition and how they relate to protease inhibition. RESULTS: The crystal structure of a mutant form of human PAI-2 in the stressed state has been determined at 2.0 A resolution. The RCL is completely disordered in the structure. An examination of polar residues that are highly conserved across all serpins identifies functionally important regions. A buried polar cluster beneath betasheet A (the so-called 'shutter' region) is found to stabilise both the stressed and relaxed forms via a rearrangement of hydrogen bonds. CONCLUSIONS: A statistical analysis of interstrand interactions indicated that the shutter region can be used to discriminate between inhibitory and non-inhibitory serpins. This analysis implied that insertion of the RCL into betasheet A up to residue P8 is important for protease inhibition and hence the structure of the complex formed between the serpin and the target protease. PMID- 10368273 TI - The 1.2 A crystal structure of hirustasin reveals the intrinsic flexibility of a family of highly disulphide-bridged inhibitors. AB - BACKGROUND: Leech-derived inhibitors have a prominent role in the development of new antithrombotic drugs, because some of them are able to block the blood coagulation cascade. Hirustasin, a serine protease inhibitor from the leech Hirudo medicinalis, binds specifically to tissue kallikrein and possesses structural similarity with antistasin, a potent factor Xa inhibitor from Haementeria officinalis. Although the 2.4 A structure of the hirustasin kallikrein complex is known, classical methods such as molecular replacement were not successful in solving the structure of free hirustasin. RESULTS: Ab initio real/reciprocal space iteration has been used to solve the structure of free hirustasin using either 1.4 A room temperature data or 1.2 A low temperature diffraction data. The structure was also solved independently from a single pseudo-symmetric gold derivative using maximum likelihood methods. A comparison of the free and complexed structures reveals that binding to kallikrein causes a hinge-bending motion between the two hirustasin subdomains. This movement is accompanied by the isomerisation of a cis proline to the trans conformation and a movement of the P3, P4 and P5 residues so that they can interact with the cognate protease. CONCLUSIONS: The inhibitors from this protein family are fairly flexible despite being highly cross-linked by disulphide bridges. This intrinsic flexibility is necessary to adopt a conformation that is recognised by the protease and to achieve an optimal fit, such observations illustrate the pitfalls of designing inhibitors based on static lock-and-key models. This work illustrates the potential of new methods of structure solution that require less or even no prior phase information. PMID- 10368274 TI - Crystal structure of Trypanosoma cruzi trypanothione reductase in complex with trypanothione, and the structure-based discovery of new natural product inhibitors. AB - BACKGROUND: Trypanothione reductase (TR) helps to maintain an intracellular reducing environment in trypanosomatids, a group of protozoan parasites that afflict humans and livestock in tropical areas. This protective function is achieved via reduction of polyamine-glutathione conjugates, in particular trypanothione. TR has been validated as a chemotherapeutic target by molecular genetics methods. To assist the development of new therapeutics, we have characterised the structure of TR from the pathogen Trypanosoma cruzi complexed with the substrate trypanothione and have used the structure to guide database searches and molecular modelling studies. RESULTS: The TR-trypanothione-disulfide structure has been determined to 2.4 A resolution. The chemical interactions involved in enzyme recognition and binding of substrate can be inferred from this structure. Comparisons with the related mammalian enzyme, glutathione reductase, explain why each enzyme is so specific for its own substrate. A CH***O hydrogen bond can occur between the active-site histidine and a carbonyl of the substrate. This interaction contributes to enzyme specificity and mechanism by producing an electronic induced fit when substrate binds. Database searches and molecular modelling using the substrate as a template and the active site as receptor have identified a class of cyclic-polyamine natural products that are novel TR inhibitors. CONCLUSIONS: The structure of the TR-trypanothione enzyme-substrate complex provides details of a potentially valuable drug target. This information has helped to identify a new class of enzyme inhibitors as novel lead compounds worthy of further development in the search for improved medicines to treat a range of parasitic infections. PMID- 10368275 TI - The crystal structure of the DNase domain of colicin E7 in complex with its inhibitor Im7 protein. AB - BACKGROUND: Colicin E7 (ColE7) is one of the bacterial toxins classified as a DNase-type E-group colicin. The cytotoxic activity of a colicin in a colicin producing cell can be counteracted by binding of the colicin to a highly specific immunity protein. This biological event is a good model system for the investigation of protein recognition. RESULTS: The crystal structure of a one-to one complex between the DNase domain of colicin E7 and its cognate immunity protein Im7 has been determined at 2.3 A resolution. Im7 in the complex is a varied four-helix bundle that is identical to the structure previously determined for uncomplexed Im7. The structure of the DNase domain of ColE7 displays a novel alpha/beta fold and contains a Zn2+ ion bound to three histidine residues and one water molecule in a distorted tetrahedron geometry. Im7 has a V-shaped structure, extending two arms to clamp the DNase domain of ColE7. One arm (alpha1(*)-loop12 alpha2(*); where * represents helices in Im7) is located in the region that displays the greatest sequence variation among members of the immunity proteins in the same subfamily. This arm mainly uses acidic sidechains to interact with the basic sidechains in the DNase domain of ColE7. The other arm (loop 23 alpha3(*)-loop 34) is more conserved and it interacts not only with the sidechain but also with the mainchain atoms of the DNase domain of ColE7. CONCLUSIONS: The protein interfaces between the DNase domain of ColE7 and Im7 are charge complementary and charge interactions contribute significantly to the tight and specific binding between the two proteins. The more variable arm in Im7 dominates the binding specificity of the immunity protein to its cognate colicin. Biological and structural data suggest that the DNase active site for ColE7 is probably near the metal-binding site. PMID- 10368276 TI - The structure of active serpin 1K from Manduca sexta. AB - BACKGROUND: The reactive center loops (RCL) of serpins undergo large conformational changes triggered by the interaction with their target protease. Available crystallographic data suggest that the serpin RCL is polymorphic, but the relevance of the observed conformations to the competent active structure and the conformational changes that occur on binding target protease has remained obscure. New high-resolution data on an active serpin, serpin 1K from the moth hornworm Manduca sexta, provide insights into how active serpins are stabilized and how conformational changes are induced by protease binding. RESULTS: The 2.1 A structure shows that the RCL of serpin 1K, like that of active alpha1 antitrypsin, is canonical, complimentary and ready to bind to the target protease between P3 and P3 (where P refers to standard protease nomenclature),. In the hinge region (P17-P13), however, the RCL of serpin 1K, like ovalbumin and alpha1 antichymotrypsin, forms tight interactions that stabilize the five-stranded closed form of betasheet A. These interactions are not present in, and are not compatible with, the observed structure of active alpha1-antitrypsin. CONCLUSIONS: Serpin 1K may represent the best resting conformation for serpins - canonical near P1, but stabilized in the closed conformation of betasheet A. By comparison with other active serpins, especially alpha1-antitrypsin, a model is proposed in which interaction with the target protease near P1 leads to conformational changes in betasheet A of the serpin. PMID- 10368277 TI - A firm hand on NFkappaB: structures of the IkappaBalpha-NFkappaB complex. AB - Two crystal structures of an IkappaB-NFkappaB complex have recently been determined. The structures show in detail how IkappaB controls the subcellular localization and activity of the eukaryotic transcription factor NFkappaB. PMID- 10368278 TI - The art of matchmaking: sequence alignment methods and their structural implications. PMID- 10368279 TI - The active conformation of plasminogen activator inhibitor 1, a target for drugs to control fibrinolysis and cell adhesion. AB - BACKGROUND: Plasminogen activator inhibitor 1 (PAI-1) is a serpin that has a key role in the control of fibrinolysis through proteinase inhibition. PAI-1 also has a role in regulating cell adhesion processes relevant to tissue remodeling and metastasis; this role is mediated by its binding to the adhesive glycoprotein vitronectin rather than by proteinase inhibition. Active PAI-1 is metastable and spontaneously transforms to an inactive latent conformation. Previous attempts to crystallize the active conformation of PAI-1 have failed. RESULTS: The crystal structure of a stable quadruple mutant of PAI-1(Asn150-->His, Lys154-->Thr, Gln319-->Leu, Met354-->Ile) in its active conformation has been solved at a nominal 3 A resolution. In two of four independent molecules within the crystal, the flexible reactive center loop is unconstrained by crystal-packing contacts and is disordered. In the other two molecules, the reactive center loop forms intimate loop-sheet interactions with neighboring molecules, generating an infinite chain within the crystal. The overall conformation resembles that seen for other active inhibitory serpins. CONCLUSIONS: The structure clarifies the molecular basis of the stabilizing mutations and the reduced affinity of PAI-1, on cleavage or in the latent form, for vitronectin. The infinite chain of linked molecules also suggests a new mechanism for the serpin polymerization associated with certain diseases. The results support the concept that the reactive center loop of an active serpin is flexible and has no defined conformation in the absence of intermolecular contacts. The determination of the structure of the active form constitutes an essential step for the rational design of PAI-1 inhibitors. PMID- 10368280 TI - The primary and three-dimensional structures of a nine-haem cytochrome c from Desulfovibrio desulfuricans ATCC 27774 reveal a new member of the Hmc family. AB - BACKGROUND: Haem-containing proteins are directly involved in electron transfer as well as in enzymatic functions. The nine-haem cytochrome c (9Hcc), previously described as having 12 haem groups, was isolated from cells of Desulfovibrio desulfuricans ATCC 27774, grown under both nitrate- and sulphate-respiring conditions. RESULTS: Models for the primary and three-dimensional structures of this cytochrome, containing 292 amino acid residues and nine haem groups, were derived using the multiple wavelength anomalous dispersion phasing method and refined using 1.8 A diffraction data to an R value of 17.0%. The nine haem groups are arranged into two tetrahaem clusters, with Fe-Fe distances and local protein fold similar to tetrahaem cytochromes c3, while the extra haem is located asymmetrically between the two clusters. CONCLUSIONS: This is the first known three-dimensional structure in which multiple copies of a tetrahaem cytochrome c3 like fold are present in the same polypeptide chain. Sequence homology was found between this cytochrome and the C-terminal region (residues 229-514) of the high molecular weight cytochrome c from Desulfovibrio vulgaris Hildenborough (DvH Hmc). A new haem arrangement in domains III and IV of DvH Hmc is proposed. Kinetic experiments showed that 9Hcc can be reduced by the [NiFe] hydrogenase from D. desulfuricans ATCC 27774, but that this reduction is faster in the presence of tetrahaem cytochrome c3. As Hmc has never been found in D. desulfuricans ATCC 27774, we propose that 9Hcc replaces it in this organism and is therefore probably involved in electron transfer across the membrane. PMID- 10368281 TI - Dual conformations for the HIV-1 gp120 V3 loop in complexes with different neutralizing fabs. AB - BACKGROUND: The third hypervariable (V3) loop of HIV-1 gp120 has been termed the principal neutralizing determinant (PND) of the virus and is involved in many aspects of virus infectivity. The V3 loop is required for viral entry into the cell via membrane fusion and is believed to interact with cell surface chemokine receptors on T cells and macrophages. Sequence changes in V3 can affect chemokine receptor usage, and can, therefore, modulate which types of cells are infected. Antibodies raised against peptides with V3 sequences can neutralize laboratory adapted strains of the virus and inhibit syncytia formation. Fab fragments of these neutralizing antibodies in complex with V3 loop peptides have been studied by X-ray crystallography to determine the conformation of the V3 loop. RESULTS: We have determined three crystal structures of Fab 58.2, a broadly neutralizing antibody, in complex with one linear and two cyclic peptides the amino acid sequence of which comes from the MN isolate of the gp120 V3 loop. Although the peptide conformations are very similar for the linear and cyclic forms, they differ from that seen for the identical peptide bound to a different broadly neutralizing antibody, Fab 59.1, and for a similar peptide bound to the MN specific Fab 50.1. The conformational difference in the peptide is localized around residues Gly-Pro-Gly-Arg, which are highly conserved in different HIV-1 isolates and are predicted to adopt a type II beta turn. CONCLUSIONS: The V3 loop can adopt at least two different conformations for the highly conserved Gly-Pro Gly-Arg sequence at the tip of the loop. Thus, the HIV-1 V3 loop has some inherent conformational flexibility that may relate to its biological function. PMID- 10368282 TI - The crystal structure of Cys-tRNACys-EF-Tu-GDPNP reveals general and specific features in the ternary complex and in tRNA. AB - BACKGROUND: . The translation elongation factor EF-Tu in its GTP-bound state forms a ternary complex with any aminoacylated tRNA (aa-tRNA), except initiator tRNA and selenocysteinyl-tRNA. This complex delivers aa-tRNA to the ribosomal A site during the elongation cycle of translation. The crystal structure of the yeast Phe-tRNAPhe ternary complex with Thermus aquaticus EF-Tu-GDPNP (Phe-TC) has previously been determined as one representative of this general yet highly discriminating complex formation. RESULTS: The ternary complex of Escherichia coli Cys-tRNACys and T. aquaticus EF-Tu-GDPNP (Cys-TC) has been solved and refined at 2.6 degrees resolution. Conserved and variable features of the aa-tRNA recognition and binding by EF-Tu-GTP have been revealed by comparison with the Phe-TC structure. New tertiary interactions are observed in the tRNACys structure. A 'kissing complex' is observed in the very close crystal packing arrangement. CONCLUSIONS: The recognition of Cys-tRNACys by EF-Tu-GDPNP is restricted to the aa-tRNA motif previously identified in Phe-TC and consists of the aminoacylated 3' end, the phosphorylated 5' end and one side of the acceptor stem and T stem. The aminoacyl bond is recognized somewhat differently, yet by the same primary motif in EF-Tu, which suggests that EF-Tu adapts to subtle variations in this moiety among all aa-tRNAs. New tertiary interactions revealed by the Cys-tRNACys structure, such as a protonated C16:C59 pyrimidine pair, a G15:G48 'Levitt pair' and an s4U8:A14:A46 base triple add to the generic understanding of tRNA structure from sequence. The structure of the 'kissing complex' shows a quasicontinuous helix with a distinct shape determined by the number of base pairs. PMID- 10368283 TI - Structure of a CXC chemokine-receptor fragment in complex with interleukin-8. AB - BACKGROUND: Interactions between CXC chemokines (e.g. interleukin-8, IL-8) and their receptors (e.g. CXCR-1) have a key role in host defense and disease by attracting and upregulating neutrophils to sites of inflammation. The transmembrane nature of the receptor impedes structure-based understanding of ligand interactions. Linear peptides based on the N-terminal, extracellular portion of the receptor CXCR-1 do bind to IL-8, however, and inhibit the binding of IL-8 to the full-length receptor. RESULTS: The NMR solution structure of the complex formed between IL-8 and one such receptor-based peptide indicates that a cleft between a loop and a beta hairpin constitute part of the receptor interaction surface on IL-8. Nine residues from the C terminus of the receptor peptide (corresponding to Pro21-Pro29 of CXCR-1) occupy the cleft in an extended fashion. Intermolecular contacts are mostly hydrophobic and sidechain mediated. CONCLUSIONS: The results offer the first details at an atomic level of the interaction between a chemokine and its receptor. Consideration of other biochemical data allow extrapolation to a model for the interaction of IL-8 with the full-length receptor. In this model, the heparin-binding residues of IL-8 are exposed, thereby allowing presentation of the chemokine from endothelial cell surface glycosaminoglycans. This first glimpse of how IL-8 binds to its receptor provides a foundation for the structure-based design of chemokine antagonists. PMID- 10368284 TI - The physiological structure of human C-reactive protein and its complex with phosphocholine. AB - BACKGROUND: Human C-reactive protein (CRP) is the classical acute phase reactant, the circulating concentration of which rises rapidly and extensively in a cytokine-mediated response to tissue injury, infection and inflammation. Serum CRP values are routinely measured, empirically, to detect and monitor many human diseases. However, CRP is likely to have important host defence, scavenging and metabolic functions through its capacity for calcium-dependent binding to exogenous and autologous molecules containing phosphocholine (PC) and then activating the classical complement pathway. CRP may also have pathogenic effects and the recent discovery of a prognostic association between increased CRP production and coronary atherothrombotic events is of particular interest. RESULTS: The X-ray structures of fully calcified C-reactive protein, in the presence and absence of bound PC, reveal that although the subunit beta-sheet jellyroll fold is very similar to that of the homologous pentameric protein serum amyloid P component, each subunit is tipped towards the fivefold axis. PC is bound in a shallow surface pocket on each subunit, interacting with the two protein-bound calcium ions via the phosphate group and with Glu81 via the choline moiety. There is also an unexpected hydrophobic pocket adjacent to the ligand. CONCLUSIONS: The structure shows how large ligands containing PC may be bound by CRP via a phosphate oxygen that projects away from the surface of the protein. Multipoint attachment of one planar face of the CRP molecule to a PC-bearing surface would leave available, on the opposite exposed face, the recognition sites for C1q, which have been identified by mutagenesis. This would enable CRP to target physiologically and/or pathologically significant complement activation. The hydrophobic pocket adjacent to bound PC invites the design of inhibitors of CRP binding that may have therapeutic relevance to the possible role of CRP in atherothrombotic events. PMID- 10368285 TI - Three-dimensional structure of a barley beta-D-glucan exohydrolase, a family 3 glycosyl hydrolase. AB - BACKGROUND: Cell walls of the starchy endosperm and young vegetative tissues of barley (Hordeum vulgare) contain high levels of (1-->3,1-->4)-beta-D-glucans. The (1-->3,1-->4)-beta-D-glucans are hydrolysed during wall degradation in germinated grain and during wall loosening in elongating coleoptiles. These key processes of plant development are mediated by several polysaccharide endohydrolases and exohydrolases. RESULTS: . The three-dimensional structure of barley beta-D-glucan exohydrolase isoenzyme ExoI has been determined by X-ray crystallography. This is the first reported structure of a family 3 glycosyl hydrolase. The enzyme is a two-domain, globular protein of 605 amino acid residues and is N-glycosylated at three sites. The first 357 residues constitute an (alpha/beta)8 TIM-barrel domain. The second domain consists of residues 374-559 arranged in a six-stranded beta sandwich, which contains a beta sheet of five parallel beta strands and one antiparallel beta strand, with three alpha helices on either side of the sheet. A glucose moiety is observed in a pocket at the interface of the two domains, where Asp285 and Glu491 are believed to be involved in catalysis. CONCLUSIONS: The pocket at the interface of the two domains is probably the active site of the enzyme. Because amino acid residues that line this active-site pocket arise from both domains, activity could be regulated through the spatial disposition of the domains. Furthermore, there are sites on the second domain that may bind carbohydrate, as suggested by previously published kinetic data indicating that, in addition to the catalytic site, the enzyme has a second binding site specific for (1-->3, 1-->4)-beta-D-glucans. PMID- 10368287 TI - A new proposal for urease mechanism based on the crystal structures of the native and inhibited enzyme from Bacillus pasteurii: why urea hydrolysis costs two nickels. AB - BACKGROUND: Urease catalyzes the hydrolysis of urea, the final step of organic nitrogen mineralization, using a bimetallic nickel centre. The role of the active site metal ions and amino acid residues has not been elucidated to date. Many pathologies are associated with the activity of ureolytic bacteria, and the efficiency of soil nitrogen fertilization with urea is severely decreased by urease activity. Therefore, the development of urease inhibitors would lead to a reduction of environmental pollution, to enhanced efficiency of nitrogen uptake by plants, and to improved therapeutic strategies for treatment of infections due to ureolytic bacteria. Structure-based design of urease inhibitors would require knowledge of the enzyme mechanism at the molecular level. RESULTS: The structures of native and inhibited urease from Bacillus pasteurii have been determined at a resolution of 2.0 A by synchrotron X-ray cryogenic crystallography. In the native enzyme, the coordination sphere of each of the two nickel ions is completed by a water molecule and a bridging hydroxide. A fourth water molecule completes a tetrahedral cluster of solvent molecules. The enzyme crystallized in the presence of phenylphosphorodiamidate contains the tetrahedral transition-state analogue diamidophosphoric acid, bound to the two nickel ions in an unprecedented mode. Comparison of the native and inhibited structures reveals two distinct conformations of the flap lining the active-site cavity. CONCLUSIONS: The mode of binding of the inhibitor, and a comparison between the native and inhibited urease structures, indicate a novel mechanism for enzymatic urea hydrolysis which reconciles the available structural and biochemical data. PMID- 10368286 TI - Crystal structures of Nova-1 and Nova-2 K-homology RNA-binding domains. AB - BACKGROUND: Nova-1 and Nova-2 are related neuronal proteins that were initially cloned using antisera obtained from patients with the autoimmune neurological disease paraneoplastic opsoclonus-myoclonus ataxia (POMA). Both of these disease gene products contain three RNA-binding motifs known as K-homology or KH domains, and their RNA ligands have been identified via binding-site selection experiments. The KH motif structure has been determined previously using NMR spectroscopy, but not using X-ray crystallography. Many proteins contain more than one KH domain, yet there is no published structural information regarding the behavior of such multimers. RESULTS: We have obtained the first X-ray crystallographic structures of KH-domain-containing proteins. Structures of the third KH domains (KH3) of Nova-1 and Nova-2 were determined by multiple isomorphous replacement and molecular replacement at 2.6 A and 2.0 A, respectively. These highly similar RNA-binding motifs form a compact protease resistant domain resembling an open-faced sandwich, consisting of a three stranded antiparallel beta sheet topped by three alpha helices. In both Nova crystals, the lattice is composed of symmetric tetramers of KH3 domains that are created by two dimer interfaces. CONCLUSIONS: The crystal structures of both Nova KH3 domains are similar to the previously determined NMR structures. The most significant differences among the KH domains involve changes in the positioning of one or more of the alpha helices with respect to the betasheet, particularly in the NMR structure of the KH1 domain of the Fragile X disease protein FMR-1. Loop regions in the KH domains are clearly visible in the crystal structure, unlike the NMR structures, revealing the conformation of the invariant Gly-X-X Gly segment that is thought to participate in RNA-binding and of the variable region. The tetrameric arrangements of the Nova KH3 domains provide insights into how KH domains may interact with each other in proteins containing multiple KH motifs. PMID- 10368288 TI - The solution structure of the guanine nucleotide exchange domain of human elongation factor 1beta reveals a striking resemblance to that of EF-Ts from Escherichia coli. AB - BACKGROUND: In eukaryotic protein synthesis, the multi-subunit elongation factor 1 (EF-1) plays an important role in ensuring the fidelity and regulating the rate of translation. EF-1alpha, which transports the aminoacyl tRNA to the ribosome, is a member of the G-protein superfamily. EF-1beta regulates the activity of EF 1alpha by catalyzing the exchange of GDP for GTP and thereby regenerating the active form of EF-1alpha. The structure of the bacterial analog of EF-1alpha, EF Tu has been solved in complex with its GDP exchange factor, EF-Ts. These structures indicate a mechanism for GDP-GTP exchange in prokaryotes. Although there is good sequence conservation between EF-1alpha and EF-Tu, there is essentially no sequence similarity between EF-1beta and EF-Ts. We wished to explore whether the prokaryotic exchange mechanism could shed any light on the mechanism of eukaryotic translation elongation. RESULTS: Here, we report the structure of the guanine-nucleotide exchange factor (GEF) domain of human EF 1beta (hEF-1beta, residues 135-224); hEF-1beta[135-224], determined by nuclear magnetic resonance spectroscopy. Sequence conservation analysis of the GEF domains of EF-1 subunits beta and delta from widely divergent organisms indicates that the most highly conserved residues are in two loop regions. Intriguingly, hEF-1beta[135-224] shares structural homology with the GEF domain of EF-Ts despite their different primary sequences. CONCLUSIONS: On the basis of both the structural homology between EF-Ts and hEF-1beta[135-224] and the sequence conservation analysis, we propose that the mechanism of guanine-nucleotide exchange in protein synthesis has been conserved in prokaryotes and eukaryotes. In particular, Tyr181 of hEF-1beta[135-224] appears to be analogous to Phe81 of Escherichia coli EF-Ts. PMID- 10368290 TI - The hexamerization domain of N-ethylmaleimide-sensitive factor: structural clues to chaperone function. AB - The hexameric structure of the D2 ATP-binding module of N-ethylmaleimide sensitive factor (NSF), a chaperone involved in SNARE complex disassembly, was recently determined. This structure and the previously determined structure of the DNA polymerase III delta' subunit have far-reaching biological significance because these modules are related to diverse ATPases that promote the assembly, disassembly and operation of various protein complexes. PMID- 10368289 TI - A six-stranded double-psi beta barrel is shared by several protein superfamilies. AB - BACKGROUND: Six-stranded beta barrels with a pseudo-twofold axis are found in several proteins. One group comprises a Greek-key structure with all strands antiparallel; an example is the N-terminal domain of ferredoxin reductase. Others involve parallel strands forming two psi structures (the double-psi beta barrel). A recently discovered example of the latter class is aspartate-alpha decarboxylase (ADC) from Escherichia coli, a pyruvoyl-dependent tetrameric enzyme involved in the synthesis of pantothenate. RESULTS: Visual inspection and automated database searches identified the six-stranded double-psi beta barrel in ADC, Rhodobacter sphaeroides dimethylsulfoxide (DMSO) reductase, E. coli formate dehydrogenase H (FDHH), the plant defense protein barwin, Humicola insolens endoglucanase V (EGV) and, with a circular permutation, in the aspartic proteinases. Structure-based sequence alignments revealed several interactions including hydrophobic contacts or sidechain-mainchain hydrogen bonds that position the middle beta strand under a psi loop, which may significantly contribute to stabilizing the fold. The identification of key interactions allowed the filtering of weak sequence similarities to some of these proteins, which had been detected by sequence database searches. This led to the prediction of the double-psi beta-barrel domain in several families of proteins in eukaryotes and archaea. CONCLUSIONS: The structure comparison and clustering study of double-psi beta barrels suggests that there could be a common homodimeric ancestor to ADC, FDHH and DMSO reductase, and also to barwin and EGV. There are other protein families with unknown structure that are likely to adopt the same fold. In the known structures, the protein active sites cluster around the psi loop, indicating that its rigidity, protrusion and free mainchain functional groups may be well suited to providing a framework for catalysis. PMID- 10368291 TI - Protein crystals and their evil twins. AB - Different types of crystal twinning are reviewed with an emphasis on how to detect the phenomenon from protein diffraction data. The recent literature and a database survey both serve as reminders to perform routine checks whenever twinning is a possibility. PMID- 10368292 TI - An open and closed case for all polymerases. AB - The recently determined structures of HIV-1 reverse transcriptase and Taq DNA polymerase in complex with DNA primer-template and an incoming nucleotide have shown that a large conformational change configures the polymerase active site for nucleotidyl transfer. The structure of reverse transcriptase in the catalytic complex will open the path to the rational design of novel nucleoside analog inhibitors of viral replication. PMID- 10368293 TI - X-ray structure of pyrrolidone carboxyl peptidase from the hyperthermophilic archaeon Thermococcus litoralis. AB - BACKGROUND: Pyrrolidone carboxyl peptidases (pcps) are a group of exopeptidases responsible for the hydrolysis of N-terminal pyroglutamate residues from peptides and proteins. The bacterial and archaeal pcps are members of a conserved family of cysteine proteases. The pcp from the hyperthermophilic archaeon Thermococcus litoralis is more thermostable than the bacterial enzymes with which it has up to 40% sequence identity. The pcp activity in archaea and eubacteria is proposed to be involved in detoxification processes and in nutrient metabolism; eukaryotic counterparts of the enzyme are involved in the processing of biologically active peptides. RESULTS: The crystal structure of pcp has been determined by multiple isomorphous replacement techniques at 1.73 A resolution and refined to an R factor of 18.7% (Rfree = 21.4%). The enzyme is a homotetramer of single open alpha/beta domain subunits, with a prominent hydrophobic core formed from loops coming together from each monomer. The active-site residues have been identified as a Cys143-His167-Glu80 catalytic triad. Structural homology to enzymes of different specificity and mechanism has been identified. CONCLUSIONS: The Thermococcus pcp has no sequence or structural homology with other members of the cysteine protease family. It does, however, show considerable similarities to other hydrolytic enzymes of widely varying substrate specificity and mechanism, suggesting that they are the products of divergent evolution from a common ancestor. The enhanced thermostability of the T. litoralis pcp may arise from hydrophobic interactions between the subunits and the presence of intersubunit disulphide bridges. PMID- 10368294 TI - Crystal structure of a viral cyclin, a positive regulator of cyclin-dependent kinase 6. AB - BACKGROUND: Cyclin-dependent kinases (CDKs) have a central role in cell-cycle control and are activated by complex formation with positive regulatory proteins called cyclins and by phosphorylation. The overexpression and mutation of cyclins and CDKs has been associated with tumorigenesis and oncogenesis. A virus-encoded cyclin (v-cyclin) from herpesvirus saimiri has been shown to exhibit highest sequence homology to type D cyclins and specifically activates CDK6 of host cells to a very high degree. RESULTS: We have determined the first X-ray structure of a v-cyclin to 3.0 A resolution. The structure of the core domains is very similar to those of cyclin A and cyclin H from human cells. To understand the structural basis for the v-cyclin specificity for CDK6 and the insensitivity of the complex to inhibitors of the p21 and INK4 families, a v-cyclin-CDK2 model was built on the basis of the known structures of human cyclin A in complex with CDK2 and the CDK inhibitor p27(Kip1). CONCLUSIONS: Although many critical interactions between cyclin A and CDK2 would be conserved in a v-cyclin-CDK2 complex, some appear sterically or electrostatically unfavorable due to shifts in the backbone conformation or sidechain differences and may contribute to v-cyclin selectivity for CDK6. The insensitivity of v-cyclin-CDK6 complexes to inhibitors of the p21 family is probably due to structural changes in v-cyclin that lead to a flatter surface area offering fewer potential contacts with the protein inhibitor. In addition, sequence changes in v-cyclin eliminate hydrogen-bonding partners for atoms of the p27(Kip1) inhibitor. This structure provides the first model for interactions between v-cyclins and host cell-cycle proteins; these interactions may be important for virus survival as well as oncogenic transformation of host cells. PMID- 10368295 TI - Crystal structure of the trimeric alpha-helical coiled-coil and the three lectin domains of human lung surfactant protein D. AB - BACKGROUND: Human lung surfactant protein D (hSP-D) belongs to the collectin family of C-type lectins and participates in the innate immune surveillance against microorganisms in the lung through recognition of carbohydrate ligands present on the surface of pathogens. The involvement of this protein in innate immunity and the allergic response make it the subject of much interest. RESULTS: We have determined the crystal structure of a trimeric fragment of hSP-D at 2.3 A resolution. The structure comprises an alpha-helical coiled-coil and three carbohydrate-recognition domains (CRDs). An interesting deviation from symmetry was found in the projection of a single tyrosine sidechain into the centre of the coiled-coil; the asymmetry of this residue influences the orientation of one of the adjacent CRDs. The cleft between the three CRDs presents a large positively charged surface. CONCLUSIONS: The fold of the CRD of hSP-D is similar to that of the mannan-binding protein (MBP), but its orientation relative to the alpha helical coiled-coil region differs somewhat to that seen in the MBP structure. The novel central packing of the tyrosine sidechain within the coiled-coil and the resulting asymmetric orientation of the CRDs has unexpected functional implications. The positively charged surface might facilitate binding to negatively charged structures, such as lipopolysaccharides. PMID- 10368296 TI - A 30-angstrom-long U-shaped catalytic tunnel in the crystal structure of polyamine oxidase. AB - BACKGROUND: Polyamines are essential for cell growth and differentiation; compounds interfering with their metabolism are potential anticancer agents. Polyamine oxidase (PAO) plays a central role in polyamine homeostasis. The enzyme utilises an FAD cofactor to catalyse the oxidation of the secondary amino groups of spermine and spermidine. RESULTS: The first crystal structure of a polyamine oxidase has been determined to a resolution of 1.9 Angstroms. PAO from Zea mays contains two domains, which define a remarkable 30 Angstrom long U-shaped catalytic tunnel at their interface. The structure of PAO in complex with the inhibitor MDL72527 reveals the residues forming the catalytic machinery and unusual enzyme-inhibitor CH.O H bonds. A ring of glutamate and aspartate residues surrounding one of the two tunnel openings contributes to the steering of the substrate towards the inside of the tunnel. CONCLUSIONS: PAO specifically oxidizes substrates that have both primary and secondary amino groups. The complex with MDL72527 shows that the primary amino groups are essential for the proper alignment of the substrate with respect to the flavin. Conservation of an N-terminal sequence motif indicates that PAO is member of a novel family of flavoenzymes. Among these, monoamine oxidase displays significant sequence homology with PAO, suggesting a similar overall folding topology. PMID- 10368297 TI - The structure of a Staphylococcus aureus leucocidin component (LukF-PV) reveals the fold of the water-soluble species of a family of transmembrane pore-forming toxins. AB - BACKGROUND: Leucocidins and gamma-hemolysins are bi-component toxins secreted by Staphylococcus aureus. These toxins activate responses of specific cells and form lethal transmembrane pores. Their leucotoxic and hemolytic activities involve the sequential binding and the synergistic association of a class S and a class F component, which form hetero-oligomeric complexes. The components of each protein class are produced as non-associated, water-soluble proteins that undergo conformational changes and oligomerization after recognition of their cell targets. RESULTS: The crystal structure of the monomeric water-soluble form of the F component of Panton-Valentine leucocidin (LukF-PV) has been solved by the multiwavelength anomalous dispersion (MAD) method and refined at 2.0 A resolution. The core of this three-domain protein is similar to that of alpha hemolysin, but significant differences occur in regions that may be involved in the mechanism of pore formation. The glycine-rich stem, which undergoes a major rearrangement in this process, forms an additional domain in LukF-PV. The fold of this domain is similar to that of the neurotoxins and cardiotoxins from snake venom. CONCLUSIONS: The structure analysis and a multiple sequence alignment of all toxic components, suggest that LukF-PV represents the fold of any water soluble secreted protein in this family of transmembrane pore-forming toxins. The comparison of the structures of LukF-PV and alpha-hemolysin provides some insights into the mechanism of transmembrane pore formation for the bi-component toxins, which may diverge from that of the alpha-hemolysin heptamer. PMID- 10368298 TI - The three-dimensional structure of a DNA translocating machine at 10 A resolution. AB - BACKGROUND: Head-tail connectors are viral substructures that are very important in the viral morphogenetic cycle, having roles in the formation of the precursor capsid (prohead), DNA packaging, tail binding to the mature head and in the infection process. Structural information on the connector would, therefore, help us to understand how this structure is related to a multiplicity of functions. RESULTS: Recombinant bacteriophage phi29 connectors have been crystallized in two dimensional aggregates. An average projection image and a three-dimensional map have been obtained at 8 A and 10 A resolution, respectively, from untilted and tilted images of vitrified specimens of the two-dimensional crystals. The average projection image reveals a central mass surrounding a channel with 12 appendages protruding from the central mass. The three-dimensional map reveals a wide domain surrounded by 12 appendages that interact with the prohead vertex, and a narrow domain that interacts with the bacteriophage tail. At the junction of the two domains, 12 smaller appendages are visualized. A channel runs along the axis of the connector structure and is sufficiently wide to allow a double-stranded DNA molecule to pass through. CONCLUSIONS: The propeller-like structure of the phi29 connector strengthens the notion of the connector rotating during DNA packaging. The groove formed by the two lanes of large and small appendages may act as a rail to prevent the liberation of the connector from the prohead vertex during rotation. PMID- 10368299 TI - Structure of acetylcholinesterase complexed with E2020 (Aricept): implications for the design of new anti-Alzheimer drugs. AB - BACKGROUND: Several cholinesterase inhibitors are either being utilized for symptomatic treatment of Alzheimer's disease or are in advanced clinical trials. E2020, marketed as Aricept, is a member of a large family of N-benzylpiperidine based acetylcholinesterase (AChE) inhibitors developed, synthesized and evaluated by the Eisai Company in Japan. These inhibitors were designed on the basis of QSAR studies, prior to elucidation of the three-dimensional structure of Torpedo californica AChE (TcAChE). It significantly enhances performance in animal models of cholinergic hypofunction and has a high affinity for AChE, binding to both electric eel and mouse AChE in the nanomolar range. RESULTS: Our experimental structure of the E2020-TcAChE complex pinpoints specific interactions responsible for the high affinity and selectivity demonstrated previously. It shows that E2020 has a unique orientation along the active-site gorge, extending from the anionic subsite of the active site, at the bottom, to the peripheral anionic site, at the top, via aromatic stacking interactions with conserved aromatic acid residues. E2020 does not, however, interact directly with either the catalytic triad or the 'oxyanion hole', but only indirectly via solvent molecules. CONCLUSIONS: Our study shows, a posteriori, that the design of E2020 took advantage of several important features of the active-site gorge of AChE to produce a drug with both high affinity for AChE and a high degree of selectivity for AChE versus butyrylcholinesterase (BChE). It also delineates voids within the gorge that are not occupied by E2020 and could provide sites for potential modification of E2020 to produce drugs with improved pharmacological profiles. PMID- 10368300 TI - The crystal structure of methylglyoxal synthase from Escherichia coli. AB - BACKGROUND: The reaction mechanism of methylglyoxal synthase (MGS) is believed to be similar to that of triosephosphate isomerase (TIM). Both enzymes utilise dihydroxyacetone phosphate (DHAP) to form an enediol(ate) phosphate intermediate as the first step of their reaction pathways. However, the second catalytic step in the MGS reaction pathway is characterized by the elimination of phosphate and collapse of the enediol(ate) to form methylglyoxal instead of reprotonation to form the isomer glyceraldehyde 3-phosphate. RESULTS: The crystal structure of MGS bound to formate and substoichiometric amounts of phosphate in the space group P6522 has been determined at 1.9 A resolution. This structure shows that the enzyme is a homohexamer composed of interacting five-stranded beta/alpha proteins, rather than the hallmark alpha/beta barrel structure of TIM. The conserved residues His19, Asp71, and His98 in each of the three monomers in the asymmetric unit bind to a formate ion that is present in the crystallization conditions. Differences in the three monomers in the asymmetric unit are localized at the mouth of the active site and can be ascribed to the presence or absence of a bound phosphate ion. CONCLUSIONS: In agreement with site-directed mutagenesis and mechanistic enzymology, the structure suggests that Asp71 acts as the catalytic base. Further, Asp20 and Asp101 are involved in intersubunit salt bridges. These salt bridges may provide a pathway for transmitting allosteric information. PMID- 10368301 TI - Crystal structure of the kinase domain of human vascular endothelial growth factor receptor 2: a key enzyme in angiogenesis. AB - BACKGROUND: Angiogenesis is involved in tumor growth, macular degeneration, retinopathy and other diseases. Vascular endothelial growth factor (VEGF) stimulates angiogenesis by binding to specific receptors (VEGFRs) on the surface of vascular endothelial cells. VEGFRs are receptor tyrosine kinases that, like the platelet-derived growth factor receptors (PDGFRs), contain a large insert within the kinase domain. RESULTS: We report here the generation, kinetic characterization, and 2.4 A crystal structure of the catalytic kinase domain of VEGF receptor 2 (VEGFR2). This protein construct, which lacks 50 central residues of the 68-residue kinase insert domain (KID), has comparable kinase activity to constructs containing the entire KID. The crystal structure, determined in an unliganded phosphorylated state, reveals an overall fold and catalytic residue positions similar to those observed in other tyrosine-kinase structures. The kinase activation loop, autophosphorylated on Y1059 prior to crystallization, is mostly disordered; however, a portion of it occupies a position inhibitory to substrate binding. The ends of the KID form a beta-like structure, not observed in other known tyrosine kinase structures, that packs near to the kinase C terminus. CONCLUSIONS: The majority of the VEGFR2 KID residues are not necessary for kinase activity. The unique structure observed for the ends of the KID may also occur in other PDGFR family members and may serve to properly orient the KID for signal transduction. This VEGFR2 kinase structure provides a target for design of selective anti-angiogenic therapeutic agents. PMID- 10368303 TI - Design of sequences with good folding properties in coarse-grained protein models. AB - BACKGROUND: Designing amino acid sequences that are stable in a given target structure amounts to maximizing a conditional probability. A straightforward approach to accomplishing this is a nested Monte Carlo where the conformation space is explored over and over again for different fixed sequences; this requires excessive computational demand. Several approximate attempts to remedy this situation, based on energy minimization for fixed structure or high-T expansions, have been proposed. These methods are fast but often not accurate, as folding occurs at low T. RESULTS: We have developed a multisequence Monte Carlo procedure where both sequence and conformational space are simultaneously probed with efficient prescriptions for pruning sequence space. The method is explored on hydrophobic/polar models. First we discuss short lattice chains in order to compare with exact data and with other methods. The method is then successfully applied to lattice chains with up to 50 monomers and to off-lattice 20mers. CONCLUSIONS: The multisequence Monte Carlo method offers a new approach to sequence design in coarse-grained models. It is much more efficient than previous Monte Carlo methods, and is, as it stands, applicable to a fairly wide range of two-letter models. PMID- 10368302 TI - Monomeric sarcosine oxidase: structure of a covalently flavinylated amine oxidizing enzyme. AB - BACKGROUND: Monomeric sarcosine oxidases (MSOXs) are among the simplest members of a recently recognized family of eukaryotic and prokaryotic enzymes that catalyze similar oxidative reactions with various secondary or tertiary amino acids and contain covalently bound flavins. Other members of this family include heterotetrameric sarcosine oxidase, N-methyltryptophan oxidase and pipecolate oxidase. Mammalian sarcosine dehydrogenase and dimethylglycine dehydrogenase may be more distantly related family members. RESULTS: The X-ray crystal structure of MSOX from Bacillus sp. B-0618, expressed in Escherichia coli, has been solved at 2.0 A resolution by multiwavelength anomalous dispersion (MAD) from crystals of the selenomethionine-substituted enzyme. Fourteen selenium sites, belonging to two MSOX molecules in the asymmetric unit, were used for MAD phasing and to define the local twofold symmetry axis for electron-density averaging. The structures of the native enzyme and of two enzyme-inhibitor complexes were also determined. CONCLUSIONS: MSOX is a two-domain protein with an overall topology most similar to that of D-amino acid oxidase, with which it shares 14% sequence identity. The flavin ring is located in a very basic environment, making contact with sidechains of arginine, lysine, histidine and the N-terminal end of a helix dipole. The flavin is covalently attached through an 8alpha-S-cysteinyl linkage to Cys315 of the catalytic domain. Covalent attachment is probably self-catalyzed through interactions with the positive sidechains and the helix dipole. Substrate binding is probably stabilized by hydrogen bonds between the substrate carboxylate and two basic sidechains, Arg52 and Lys348, located above the re face of the flavin ring. PMID- 10368304 TI - Courageous science: structural studies of bluetongue virus core. AB - The structure of the bluetongue virus core was recently reported and represents the largest structure determined to atomic resolution. As a biological machine capable of RNA transcription, the structure has immense biological significance. PMID- 10368305 TI - High energy exchange: proteins that make or break phosphoramidate bonds. AB - Several proteins that catalyze phosphoryl transfer reactions involving phosphohistidine residues have recently been structurally characterized. The architecture of two histidine kinases defines a new protein kinase fold. The diverse folds of several phosphotransfer proteins appear to be designed to foster protein-protein interactions between transfer partners. PMID- 10368306 TI - New tricks for modelers from the crystallography toolkit: the particle mesh Ewald algorithm and its use in nucleic acid simulations. PMID- 10368307 TI - Crystal structure of the first dissimilatory nitrate reductase at 1.9 A solved by MAD methods. AB - BACKGROUND: The periplasmic nitrate reductase (NAP) from the sulphate reducing bacterium Desulfovibrio desulfuricans ATCC 27774 is induced by growth on nitrate and catalyses the reduction of nitrate to nitrite for respiration. NAP is a molybdenum-containing enzyme with one bis-molybdopterin guanine dinucleotide (MGD) cofactor and one [4Fe-4S] cluster in a single polypeptide chain of 723 amino acid residues. To date, there is no crystal structure of a nitrate reductase. RESULTS: The first crystal structure of a dissimilatory (respiratory) nitrate reductase was determined at 1.9 A resolution by multiwavelength anomalous diffraction (MAD) methods. The structure is folded into four domains with an alpha/beta-type topology and all four domains are involved in cofactor binding. The [4Fe-4S] centre is located near the periphery of the molecule, whereas the MGD cofactor extends across the interior of the molecule interacting with residues from all four domains. The molybdenum atom is located at the bottom of a 15 A deep crevice, and is positioned 12 A from the [4Fe-4S] cluster. The structure of NAP reveals the details of the catalytic molybdenum site, which is coordinated to two MGD cofactors, Cys140, and a water/hydroxo ligand. A facile electron-transfer pathway through bonds connects the molybdenum and the [4Fe-4S] cluster. CONCLUSIONS: The polypeptide fold of NAP and the arrangement of the cofactors is related to that of Escherichia coli formate dehydrogenase (FDH) and distantly resembles dimethylsulphoxide reductase. The close structural homology of NAP and FDH shows how small changes in the vicinity of the molybdenum catalytic site are sufficient for the substrate specificity. PMID- 10368308 TI - Modulation of the toxicity and macromolecular binding of benzene metabolites by NAD(P)H:Quinone oxidoreductase in transfected HL-60 cells. AB - Benzene is oxidized in the liver to produce a series of hydroxylated metabolites, including hydroquinone and 1,2,4-benzenetriol. These metabolites are activated to toxic and genotoxic species in the bone marrow via oxidation by myeloperoxidase (MPO). NAD(P)H:quinone oxidoreductase (NQO1) is an enzyme capable of reducing the oxidized quinone metabolites and thereby potentially reducing their toxicities. We introduced the NQO1 gene into the HL-60 cell line to create a high MPO-, high NQO1-expressing cell line, and tested its response in assays of benzene metabolite toxicity. NQO1 expression reduced a class of hydroquinone- and benzenetriol-induced DNA adducts by 79-86%. The cytotoxicity and apoptosis caused by hydroquinone were modestly reduced, while protein binding was unchanged and the rate of glutathione depletion increased. NQO1's activity in reducing a class of benzene metabolite-induced DNA adducts may be related to its known activities in maintaining membrane-bound endogenous antioxidants in reduced form. Alternatively, NQO1 activity may prevent the formation of adducts which result from polymerized products of the quinones. In either case, this protection by NQO1 may be an important mechanism in the observation that a lack of NQO1 activity affords an increased risk of benzene poisoning in exposed individuals [Rothman, N., et al. (1997) Cancer Res. 57, 2839-2842]. PMID- 10368309 TI - A simple and sensitive method for in vitro quantitation of abasic sites in DNA. AB - A novel method for the quantitation of abasic sites (AP sites) in DNA is described. As abasic sites can be generated by controlled thermal treatment of base-modified DNA, this method can be used for estimation of the extent of DNA damage resulting from exposure to genotoxic agents. The method involves use of probe molecules 1 and 2 that contain a fluorescent label linked to an aminooxy group which reacts specifically with the aldehydic function of the ring-opened form of abasic sites. The two fluorescent probes 1 and 2 were found to react with 2-deoxyribose, a model substrate, at the optimum of pH 4.0. As spontaneous depurination occurs at low pH, the reactions with abasic DNA were carried out at neutral pH with an excess concentration of the probes. Studies with alkylated, depurinated calf thymus DNA showed that the method is selective and quantitative. Good correlations were found between the level of 7-methylguanine (7-MeGua), generated in vitro in DNA by the methylating agent dimethyl sulfate, and the amount of AP sites as determined by the method presented here. In addition, similar correlations were found when the assay was used to detect abasic sites in DNA isolated from rats treated with carcinogenic alkylating agents. In each case, the level of abasic sites, as expected, is slightly higher than the level of 7 MeGua which is known to represent about 70% of the total modifications of DNA following exposure to the methylating agent. This method may be useful not only in experimental settings but also in studies of DNA damage in humans resulting from chemotherapy or exposure to environmental agents. PMID- 10368310 TI - The nutritional supplement chromium(III) tris(picolinate) cleaves DNA. AB - Chromium(III) tris(picolinate) [Cr(pic)3] is currently a very popular nutritional supplement; however, its safety has recently been questioned, especially with regard to its ability to act as a clastogen. At physiologically relevant concentrations, Cr(pic)3 is reduced by biological reductants, including ascorbate and thiols, to Cr(II)-containing species. These species are susceptible to air oxidation, resulting in the catalytic generation of the potent DNA-damaging agent hydroxyl radical. In the absence of reductants, H2O2 can interact with Cr(pic)3 to produce hydroxyl radicals by a second, less efficient mechanism. Cr(pic)3 is extremely stable, which allows the complex to be readily absorbed but also to potentially be incorporated into cells intact. In this form, Cr(pic)3 is primed by its redox potential to enter into the generation of hydroxyl radicals. This study suggests that investigation of the long-term effects of supplementation of the diet with Cr(pic)3 are needed to assess the safety of this material. PMID- 10368311 TI - Procainamide, a drug causing lupus, induces prostaglandin H synthase-2 and formation of T cell-sensitizing drug metabolites in mouse macrophages. AB - Procainamide (PA) may cause drug-induced lupus, and its reactive metabolites, hydroxylamine-PA (HAPA) and nitroso-PA, are held responsible for this. Here, we show that N-oxidation of PA to these metabolites can take place in macrophages and lead to formation of neoantigens that sensitize T cells. Murine peritoneal macrophages (PMvarphi), exposed to PA in vitro, generated neoantigens related to HAPA as indicated by (1) their capacity to elicit a specific recall response of HAPA-primed T cells in the adoptive transfer popliteal lymph node (PLN) assay and (2) the appearance of metabolite-bound protein in PA-pulsed PMvarphi, as determined by Western blot. Analysis of five phase I enzymes that might be responsible for HAPA formation by PMvarphi pointed to prostaglandin H synthase-2 (PGHS-2) as a likely candidate. Experimental evidence that PA can be oxidized to HAPA by PGHS was obtained by exposing PA to PGHS in vitro. The resulting metabolites were identified by mass spectral analysis and covalent protein binding in ELISA. In vitro, PA exposure of PMvarphi of slow acetylator A/J and fast acetylator C57BL/6 mice failed to show significant strain differences in enzyme mRNA expression, enzyme activities, or formation of HAPA-related neoantigens. By contrast, after long-term PA treatment in vivo only in slow acetylators the PMvarphi harbored HAPA-related neoantigens and T cells were sensitized to them. PMvarphi of fast acetylator C57BL/6 mice only contained HAPA related neoantigens, and their T cells were only sensitized to them if, in addition to long-term PA treatment, their donors had received injections of phorbol myristate acetate (PMA), a known enhancer of oxidative enzymes in phagocytes. In conclusion, PA treatment leads to N-oxidation of PA by enzymes, in particular PGHS-2, present in antigen-presenting cells (APC) and, hence, to generation of neoantigens which sensitize T cells. The enhanced neoantigen formation and T cell sensitization seen in slow acetylators might be explained by their higher concentration of PA substrate that is available for extrahepatic N oxidation in APC. PMID- 10368312 TI - Mutagenicity of site-specifically located 1,N2-ethenoguanine in Chinese hamster ovary cell chromosomal DNA. AB - The adduct 1,N2-etheno(epsilon)-guanine (Gua) can be formed in DNA from exogenous or endogenous bifunctional electrophiles. Previous work with site-specifically modified oligonucleotides has shown all three possible base substitutions at the site of this residue in bacterial cells and in primer extension assays using purified polymerases (with the purified polymerases also showing deletions). A 10 mer was synthesized containing 1,N2-epsilon-Gua at a specific position and ligated into a modified pCNheIA vector, which was used to insert the modified sequence into the chromosomes of AA8 (wild-type) and UV5 (nucleotide excision repair-deficient) Chinese hamster ovary cells. Transformants were selected by antibiotic resistance; DNA was amplified by polymerase chain reaction, and resistance to the restriction endonuclease NheI was used to estimate mutation frequency. In the AA8 cells, the apparent mutation frequency was elevated >10 fold due to the presence of 1, N2-epsilon-Gua (to 4.6%). In UV5 cells, the mutation frequency was even higher (7.8%), but the estimate of the frequency in the control system (vector and unmodified sequence only) was 4.5%. Sequence analysis of 21 clones derived from the mutant fraction yielded five that correspond to base pair mutations directly at the 1, N2-epsilon-Gua site. The remainder of the mutants differed from those generated from the unmodified oligonucleotide and included deletions, rearrangements, double mutants, and base pair substitutions at sites nearby but not at the 1,N2-epsilon-Gua site. PMID- 10368313 TI - Studies on the pyrrolinone metabolites derived from the tobacco alkaloid 1-methyl 2-(3-pyridinyl)pyrrole (beta-nicotyrine). AB - Previous studies have established that the tobacco alkaloid 1-methyl-2-(3 pyridyl)pyrrole (beta-nicotyrine) is biotransformed by rabbit lung and liver microsomal preparations to an equilibrium mixture of the corresponding 3- and 4 pyrrolin-2-ones. Autoxidation of these pyrrolin-2-ones generates the chemically stable 5-hydroxy-5-(3-pyridinyl)-3-pyrrolin-2-one. This paper summarizes efforts to document more completely the pathway leading to this hydroxypyrrolinone. Chemical and spectroscopic evidence implicates the 2-hydroxy-1-methyl-5-(3 pyridinyl)pyrrole (2-hydroxy-beta-nicotyrine) as the key intermediate in this reaction pathway. Of potential toxicological interest is the detection of radical species derived from the autoxidation of this compound. PMID- 10368315 TI - Glutathione-dependent generation of reactive oxygen species by the peroxidase catalyzed redox cycling of flavonoids. AB - Catalytic concentrations of apigenin (a flavone containing a phenol B ring) and naringin or naringenin (flavanones containing a phenol B ring) caused extensive GSH oxidation at a physiological pH in the presence of peroxidase. Only catalytic H2O2 concentrations were required, indicating a redox cycling mechanism that generated H2O2 was involved. Extensive oxygen uptake ensued, the extent of which was proportional to the extent of GSH oxidation to GSSG and was markedly increased by superoxide dismutase. These results suggest that prooxidant phenoxyl radicals formed by these flavonoids co-oxidized GSH to form thiyl radicals which activated oxygen. GSH also prevented the peroxidase-catalyzed oxidative destruction of these flavonoids which suggests that phenoxyl radicals initiated the oxidative destruction. This is the first time that a group of flavonoids have been identified as prooxidants independent of autoxidation reactions catalyzed by the transition metal ions Fe3+, Fe2+, Mn2+, and Cu2+. PMID- 10368314 TI - DNA damage in deoxynucleosides and oligonucleotides treated with peroxynitrite. AB - Peroxynitrite (ONOO-) is a powerful oxidizing agent that forms in a reaction of nitric oxide (NO*) and superoxide (O2-*). We have investigated ONOO--induced DNA damage using deoxynucleosides and oligonucleotides as model substrates, with particular attention paid to the oxidation of 8-oxodG by ONOO-. With regard to deoxynucleosides, ONOO- was found to have significant reactivity only with dG; dA, dC, and dT showed minimal reactivity. However, two of the major products of ONOO--induced oxidation of dG (8-oxodG and 8-nitroG) were both found to be significantly more reactive with ONOO- than with dG. In the context of an oligonucleotide, we observed a concentration-dependent oxidation of 8-oxodG to at least two types of products, one appearing at ONOO- concentrations of /=500 microM. We also examined the susceptibility of these oxidation products to repair by FaPy glycosylase, endonuclease III, uracil glycosylase, and MutY. FaPy glycosylase, which recognizes 8-oxoG as its primary substrate, was the only enzyme that exhibited an efficient reaction with 8-oxodG oxidation products at low ONOO- concentrations (/=500 microM either was not recognized or was poorly repaired by the enzymes. While processing of the lesions was inefficient with endonuclease III and not apparent with uracil glycosylase, the excision of A opposite an 8-oxoG lesion by the enzyme MutY was not affected by the reaction of 8-oxoG with ONOO-. In addition to demonstrating the complexity of ONOO- DNA damage chemistry, these results suggest that 8-oxodG may be a primary target of ONOO- in DNA. PMID- 10368316 TI - Reaction of peroxynitrite with melatonin: A mechanistic study. AB - The pH profile of the peroxynitrite/melatonin reaction suggests that both peroxynitrous acid (ONOOH) and its anion (ONOO-) are reactive toward melatonin, but at physiological pH most of the reaction with melatonin involves ONOOH and the activated form of peroxynitrous acid (ONOOH). The formation of hydroxylated products (mainly 6-hydroxymelatonin) suggests that melatonin also reacts with ONOOH. The overall peroxynitrite/melatonin reaction is first-order in melatonin and first-order in peroxynitrite, but the hydroxylation of melatonin is presumed to be zero-order in melatonin. Melatonin is metabolized in the liver, mainly to 6 hydroxymelatonin, so we do not think this metabolite is a useful biomarker for melatonin's antioxidant activity; however, 6-hydroxymelatonin is a better chain breaking antioxidant than melatonin and may contribute to the beneficial effects of melatonin in vivo. As is now well-known, CO2 modulates the reactions of peroxynitrite. The reaction of peroxynitrite with melatonin in the absence of added bicarbonate produces mainly 6-hydroxymelatonin and 1,2,3,3a,8, 8a-hexahydro 1-acetyl-5-methoxy-8a-hydroxypyrrolo[2,3-b]indole, with some isomeric 1,2,3,3a,8, 8a-hexahydro-1-acetyl-5-methoxy-3a-hydroxypyrrolo[2,3-b]indole. In the presence of added bicarbonate, product yields decrease and 6-hydroxymelatonin is not formed. These facts suggest that melatonin scavenges reactive species (such as CO3*- and *NO2) that are produced from the peroxynitrite/CO2 reaction. The spectrum of the melatoninyl radical cation is observed both in the absence and in the presence of added bicarbonate, suggesting that the melatoninyl radical cation is the initial product and the hydroxypyrrolo[2, 3-b]indole products are derived from it. Unlike tyrosine, where both nitrated and hydroxylated products can be isolated, nitromelatonin is not found in the final products from the melatonin/peroxynitrite reaction in either the absence or presence of added bicarbonate. However, we suggest that 2-hydroxy-3-nitro- and/or 2-hydroxy-3 peroxynitro-2,3-dihydromelatonin are formed as intermediates and subsequently decompose to give 1,2,3,3a,8, 8a-hexahydro-1-acetyl-5-methoxy-8a hydroxypyrrolo[2,3-b]indole. Since peroxynitrite/CO2 governs the reactions of peroxynitrite in vivo, we suggest that the hydroxypyrrolo[2,3-b]indole products are the main products from the oxidation of melatonin by peroxynitrite-derived species in vivo, and that these products may serve as indexes for melatonin's antioxidant activity. PMID- 10368317 TI - Reactions of beta-carotene with cigarette smoke oxidants. Identification of carotenoid oxidation products and evaluation of the prooxidant/antioxidant effect. AB - Recent intervention trials reported that smokers given dietary beta-carotene supplementation exhibited an increased risk of lung cancer and overall mortality. beta-Carotene has been hypothesized to promote lung carcinogenesis by acting as a prooxidant in the smoke-exposed lung. We have examined the interactions of cigarette smoke with beta-carotene in model systems. Both whole smoke and gas phase smoke oxidized beta-carotene in toluene to several products, including carbonyl-containing polyene chain cleavage products and beta-carotene epoxides. A major product of the reaction was identified as 4-nitro-beta-carotene, which was formed by nitrogen oxides in smoke. Both cis and all-trans isomers of 4-nitro beta-carotene were detected. The hypothesis that smoke-driven beta-carotene autoxidation exerts prooxidant effects was tested in a liposome system. Lipid peroxidation in dilinoleoylphosphatidylcholine liposomes exposed to gas-phase smoke was modestly inhibited by the incorporation of 0.1 mol % beta-carotene. Both the lipid soluble antioxidant alpha-tocopherol and the water soluble antioxidant ascorbate were oxidized more slowly by gas-phase smoke exposure in liposomes containing beta-carotene. These data indicate that beta-carotene exerts weak antioxidant effects against smoke-induced oxidative damage in vitro. It is unlikely that a prooxidant effect of beta-carotene occurs under biologically relevant conditions or is responsible for an increased incidence of lung cancer observed in smokers who consume beta-carotene supplements. PMID- 10368318 TI - Mammalian cells expressing Escherichia coli O6-alkylguanine-DNA alkyltransferases are hypersensitive to dibromoalkanes. AB - The effect of expression of the DNA repair protein, O6-alkylguanine-DNA alkyltransferase, on the growth inhibitory effects of the dibromoalkanes (DBA) dibromomethane (DBM) and dibromoethane (DBE) was determined in Chinese hamster lung fibroblasts transfected with and expressing high levels of the Escherichia coli alkyltransferase (ATase) genes. These included the ogt gene and complete or truncated versions of the E. coli ada gene encoding either O6-alkylguanine (O6 alkG) or alkylphosphotriester (alkPT) ATase activities. The functional activity of the ATase in these cells was demonstrated by in vitro assay of cell extracts using 3H-methylated DNA as a substrate, and by the protection they provided against the growth inhibitory effects of methylating agents N-methyl-N'-nitro-N nitrosoguanidine (MNNG) and N-methyl-N-nitrosourea (MNU) and the chloroethylating agent 1, 3-bis(2-chloroethyl)-1-nitrosourea (BCNU). However, cells expressing the full length or the O6-alkG ATase region, but not the alkPT ATase region, of Ada were found to be more sensitive to the growth inhibitory effects of the DBA; Ogt expression sensitized cells to DBM but not significantly to DBE. Addition of DBA to cell extracts depleted O6-alkG ATase activity on the methylated DNA substrate, but had no effect on alkPT ATase activity. This suggests that ATase-mediated sensitization of the intact cells may be related to the inactivation of the ATase protein. Addition to the cell culture medium of GSH or buthionine sulfoximine in attempts to augment or deplete cellular levels of GSH had no marked effect on the ATase-mediated sensitization to DBA. This suggests that rather than GSH-mediated DNA damage, the effect may be mediated by a DNA adduct caused by the oxidative metabolic pathway. These observations indicate that expression of ATase may have a detrimental effect on cellular sensitivity to environmentally relevant alkylating agents. PMID- 10368333 TI - Invited editorial on "Neuromechanical interaction in human snoring and upper airway obstruction". PMID- 10368334 TI - Neuromechanical interaction in human snoring and upper airway obstruction. AB - The fact that snoring and obstructive apnea only occur during sleep means that effective neuromuscular functioning of the upper airway during sleep is vital for the maintenance of unimpeded breathing. Recent clinical studies in humans have obtained evidence demonstrating that upper airway neural receptors sense the negative pressure generated by inspiration and "trigger," with a certain delay, reflex muscle activation to sustain the airway that might otherwise collapse. These findings have enabled us to propose a model in which the mechanics is coupled to the neuromuscular physiology through the generation of reflex wall stiffening proportional to the retarded fluid pressure. Preliminary results on this model exhibit three kinds of behavior typical of unimpeded breathing, snoring, and obstructive sleep apnea, respectively. We suggest that the increased latency of the reflex muscle activation in sleep, together with the reduced strength of the reflex, have important clinical consequences. PMID- 10368335 TI - Differential lung mechanics are genetically determined in inbred murine strains. AB - Genetic determinants of lung structure and function have been demonstrated by differential phenotypes among inbred mice strains. For example, previous studies have reported phenotypic variation in baseline ventilatory measurements of standard inbred murine strains as well as segregant and nonsegregant offspring of C3H/HeJ (C3) and C57BL/6J (B6) progenitors. One purpose of the present study is to test the hypothesis that a genetic basis for differential baseline breathing pattern is due to variation in lung mechanical properties. Quasi-static pressure volume curves were performed on standard and recombinant inbred strains to explore the interactive role of lung mechanics in determination of functional baseline ventilatory outcomes. At airway pressures between 0 and 30 cmH2O, lung volumes are significantly (P < 0.01) greater in C3 mice relative to the B6 and A/J strains. In addition, the B6C3F1/J offspring demonstrate lung mechanical properties significantly (P < 0.01) different from the C3 progenitor but not distinguishable from the B6 progenitor. With the use of recombinant inbred strains derived from C3 and B6 progenitors, cosegregation analysis between inspiratory timing and measurements of lung volume and compliance indicate that strain differences in baseline breathing pattern and pressure-volume relationships are not genetically associated. Although strain differences in lung volume and compliance between C3 and B6 mice are inheritable, this study supports a dissociation between differential inspiratory time at baseline, a trait linked to a putative genomic region on mouse chromosome 3, and differential lung mechanics among C3 and B6 progenitors and their progeny. PMID- 10368336 TI - Effect of oral glutamine on whole body carbohydrate storage during recovery from exhaustive exercise. AB - The purpose of this study was to determine the efficacy of glutamine in promoting whole body carbohydrate storage and muscle glycogen resynthesis during recovery from exhaustive exercise. Postabsorptive subjects completed a glycogen-depleting exercise protocol, then consumed 330 ml of one of three drinks, 18.5% (wt/vol) glucose polymer solution, 8 g glutamine in 330 ml glucose polymer solution, or 8 g glutamine in 330 ml placebo, and also received a primed constant infusion of [1 13C]glucose for 2 h. Plasma glutamine concentration was increased after consumption of the glutamine drinks (0.7-1.1 mM, P < 0.05). In the second hour of recovery, whole body nonoxidative glucose disposal was increased by 25% after consumption of glutamine in addition to the glucose polymer (4.48 +/- 0.61 vs. 3.59 +/- 0.18 mmol/kg, P < 0.05). Oral glutamine alone promoted storage of muscle glycogen to an extent similar to oral glucose polymer. Ingestion of glutamine and glucose polymer together promoted the storage of carbohydrate outside of skeletal muscle, the most feasible site being the liver. PMID- 10368337 TI - Dextran restores albumin-inhibited surface activity of pulmonary surfactant extract. AB - We examined the effect of dextran (molecular weight 71,000) in counteracting the surfactant inhibitory action of plasma albumin. The surface adsorption time of 0.5 mg/ml modified natural surfactant (MNS; porcine lung extract consisting of phospholipids and hydrophobic surfactant proteins) with 7.5 mg/ml albumin decreased from 681 to 143 s by addition of dextran at a concentration of 10 mg/ml (P < 0.01). The minimum surface tension of 2.0 mg/ml MNS with 30 mg/ml albumin decreased from over 21 mN/m to below 3 mN/m when dextran was added at a concentration of 10 mg/ml (P < 0.01). Surfactant-deficient newborn rabbits given 10 ml/kg of a liquid containing 2.0 mg/ml MNS with 30 mg/ml albumin had a mean tidal volume 13 ml/kg (P < 0.05). Although the underlying mechanism remains to be elucidated, we conclude that dextran restores the albumin-inhibited surface activity of MNS. PMID- 10368338 TI - Respiratory energetics during exercise at high altitude. AB - The purpose of this study was to assess the effect of high altitude (HA) on work of breathing and external work capacity. On the basis of simultaneous records of esophageal pressure and lung volume, the mechanical power of breathing (Wrs) was measured in four normal subjects during exercise at sea level (SL) and after a 1 mo sojourn at 5,050 m. Maximal exercise ventilation (VEmax) and maximal Wrs were higher at HA than at SL (mean 185 vs. 101 l/min and 129 vs. 40 cal/min, respectively), whereas maximal O2 uptake averaged 2.07 and 3.03 l/min, respectively. In three subjects, the relationship of Wrs to minute ventilation (VE) was the same at SL and HA, whereas, in one individual, Wrs for any given VE was consistently lower at HA. Assuming a mechanical efficiency (E) of 5%, the O2 cost of breathing at HA and SL should amount to 26 and 5.5% of maximal O2 uptake, whereas for E of 20% the corresponding values were 6.5 and 1.4%, respectively. Thus, at HA, Wrs may substantially limit external work unless E is high. Although at SL VEmax did not exceed the critical VE, at which any increase in VE is not useful in terms of body energetics even for E of 5%, at HA VEmax exceeded critical VE even for E of 20%. PMID- 10368339 TI - SRF protein is upregulated during stretch-induced hypertrophy of rooster ALD muscle. AB - Serum response element 1 has previously been reported to be necessary and sufficient for activation of the skeletal alpha-actin promoter during hypertrophy of the anterior latissimus dorsi (ALD) muscle of roosters [J. A. Carson, R. J. Schwartz, and F. W. Booth. Am. J. Physiol. 270 (Cell Physiol. 39): C1624-C1633, 1996]. Serum response factor (SRF) protein is the transcription factor that binds as a homodimer to serum response element 1 and activates the skeletal alpha-actin promoter. An increased expression of exogenous SRF protein in replicating C2C12 myoblasts induced a three- to fourfold activation of the skeletal alpha-actin promoter (L. Wei, W. Zhou, J. D. Croissant, F.-E. Johansen, R. Prywes, A. Balasubramamyan, and R. J. Schwartz. J. Biol. Chem. 273: 30287-30294, 1998). Thus we hypothesized that SRF protein concentration would be increased during hypertrophy of skeletal muscle. In the present study, 10% of the rooster's body weight was attached to the left wing to induce enlargement of the ALD muscle compared with the contralateral muscle. With Western analysis, a significant increase in SRF protein per gram of wet weight of the ALD muscle was noted at 7 and 13 days of hypertrophy. Furthermore, the increase in SRF protein occurred in both crude nuclear protein and cytoplasmic fractions in 7-day stretched ALD muscles. This is the first report showing increased protein concentration for a transcription factor whose regulatory element in the skeletal alpha-actin promoter has previously been shown to be required for the transduction of a hypertrophy signal in overloaded skeletal muscle of an animal. PMID- 10368340 TI - Lack of antilipolytic effect of lactate in subcutaneous abdominal adipose tissue during exercise. AB - The purpose of our study was to evaluate the potential inhibition of adipose tissue mobilization by lactate. Eight male subjects (age, 26. 25 +/- 1.75 yr) in good physical condition (maximal oxygen uptake, 59.87 +/- 2.77 ml. kg-1. min-1; %body fat, 10.15 +/- 0.89%) participated in this study. For each subject, two microdialysis probes were inserted into abdominal subcutaneous tissue. Lactate (16 mM) was perfused via one of the probes while physiological saline only was perfused via the other, both at a flow rate of 2.5 microl/min. In both probes, ethanol was also perfused for adipose tissue blood flow estimation. Dialysates were collected every 10 min during rest (30 min), exercise at 50% maximal oxygen consumption (120 min), and recovery (30 min) for the measurement of glycerol concentration. During exercise, glycerol increased significantly in both probes. However, no differences in glycerol level and ethanol extraction were observed between the lactate and control probes. These findings suggest that lactate does not impair subcutaneous abdominal adipose tissue mobilization during exercise. PMID- 10368341 TI - Ventilatory chemosensitive adaptations to intermittent hypoxic exposure with endurance training and detraining. AB - The present study was performed to clarify the effects of intermittent exposure to an altitude of 4,500 m with endurance training and detraining on ventilatory chemosensitivity. Seven subjects (sea-level group) trained at sea level at 70% maximal oxygen uptake (VO2 max) for 30 min/day, 5 days/wk for 2 wk, whereas the other seven subjects (altitude group) trained at the same relative intensity (70% altitude VO2 max) in a hypobaric chamber. VO2 max, hypoxic ventilatory response (HVR), and hypercapnic ventilatory response, as an index of central hypercapnic chemosensitivity (HCVR) and as an index of peripheral chemosensitivity (HCVRSB), were measured. In both groups VO2 max increased significantly after training, and a significant loss of VO2 max occurred during 2 wk of detraining. HVR tended to increase in the altitude group but not significantly, whereas it decreased significantly in the sea-level group after training. HCVR and HCVRSB did not change in each group. After detraining, HVR returned to the pretraining level in both groups. These results suggest that ventilatory chemosensitivity to hypoxia is more variable by endurance training and detraining than that to hypercapnia. PMID- 10368342 TI - Effect of varied extracellular PO2 on muscle performance in Xenopus single skeletal muscle fibers. AB - The purpose of this study was to examine the development of fatigue in isolated, single skeletal muscle fibers when O2 availability was reduced but not to levels considered rate limiting to mitochondrial respiration. Tetanic force was measured in single living muscle fibers (n = 6) from Xenopus laevis while being stimulated at increasing contraction rates (0.25, 0.33, 0.5, and 1 Hz) in a sequential manner, with each stimulation frequency lasting 2 min. Muscle fatigue (determined as 75% of initial maximum force) was measured during three separate work bouts (with 45 min of rest between) as the perfusate PO2 was switched between values of 30 +/- 1.9, 76 +/- 3.0, or 159 Torr in a blocked-order design. No significant differences were found in the initial peak tensions between the high-, intermediate-, and low-PO2 treatments (323 +/- 22, 298 +/- 27, and 331 +/- 24 kPa, respectively). The time to fatigue was reached significantly sooner (P < 0.05) during the 30-Torr treatment (233 +/- 39 s) compared with the 76- (385 +/- 62 s) or 159-Torr (416 +/- 65 s) treatments. The calculated critical extracellular PO2 necessary to develop an anoxic core within these fibers was 13 +/- 1 Torr, indicating that the extracellular PO2 of 30 Torr should not have been rate limiting to mitochondrial respiration. The magnitude of an unstirred layer (243 +/- 64 micron) or an intracellular O2 diffusion coefficient (0.45 +/- 0.04 x 10(-5) cm2/s) necessary to develop an anoxic core under the conditions of the study was unlikely. The earlier initiation of fatigue during the lowest extracellular PO2 condition, at physiologically high intracellular PO2 levels, suggests that muscle performance may be O2 dependent even when mitochondrial respiration is not necessarily compromised. PMID- 10368343 TI - Hyperoxia-induced changes in antioxidant capacity and the effect of dietary antioxidants. AB - We investigated, by measuring oxygen radical absorbance capacity (ORAC), whether hyperoxia causes alterations in antioxidant status and whether these alterations could be modulated by dietary antioxidants. Rats were fed for 8 wk a control diet or a control diet supplemented with vitamin E (500 IU/kg) or with aqueous extracts (ORAC: 1.36 mmol Trolox equivalents/kg) from blueberries or spinach and then were exposed to air or >99% O2 for 48 h. Although the constituents of the extracts were not extensively characterized, HPLC indicated that blueberry extract was particularly rich in anthocyanins, and the spinach extract did not contain any anthocyanins. The ORAC was determined in samples without proteins [serum treated with perchloric acid (PCA); ORACPCA] and with proteins (ORACtot). Hyperoxia induced a decrease in serum protein concentration, an increase in serum ORACPCA, decreases in lung ORACPCA and ORACtot, and an equilibration of proteins and ORACPCA between serum and pleural effusion. These alterations suggested a redistribution of antioxidants between tissues and an increase in capillary permeability during hyperoxia. Only the blueberry extract was effective in alleviating the hyperoxia-induced redistribution of antioxidants between tissues. PMID- 10368344 TI - Intrathymic and intrasplenic oxidative stress mediates thymocyte and splenocyte damage in acutely exercised mice. AB - Reactive oxygen species may contribute to apoptosis in lymphoid tissues observed after exercise. Thymic and splenic tissues excised from control mice (C) or mice immediately after (t0) or 24 h after (t24) a run to exhaustion (RTE) were assayed for biochemical indexes of oxidative stress [thymic and splenic membrane lipid peroxides, superoxide dismutase, catalase, plasma uric acid (UA), and ascorbic acid (AA)]. There were significant increases in membrane lipid peroxides in thymus (P < 0.001) and spleen (P < 0.001) in acutely exercised mice relative to controls (thymus: C = 2.74 +/- 0.80 microM; t0 = 7.45 +/- 0.48 microM; t24 = 9.44 +/-1.41 microM; spleen: C = 0.48 +/- 0.22 microM; t0 = 1.78 +/- 0.28 microM; t24 = 2. 81 +/- 0.34 microM). The thymic and splenic tissue antioxidant enzymes concentrations of superoxide dismutase and catalase were significantly lower in samples collected at t0 relative to C and t24 mice (P < 0.001). Plasma UA and AA levels were used to assess the impact of the RTE on the peripheral antioxidant pool. There was no significant change in UA levels and a significant reduction in plasma AA concentrations (P < 0.001); the reduction in plasma AA occurred at t24 (6.53 +/- 1.64 microM) relative to t0 (13.11 +/- 0. 71 microM) and C (13.26 +/- 1.2 microM). These results suggest that oxidative damage occurs in lymphoid tissues after RTE exercise and that such damage may contribute to lymphocyte damage observed after acute exercise. PMID- 10368345 TI - Impact of hyperinsulinemia on myosin heavy chain gene regulation. AB - The purpose of this study was to determine whether hyperinsulinemia alters myosin heavy chain (MHC) gene expression in human skeletal muscle. A biopsy from the vastus lateralis was obtained in young, lean [age 24.6 +/- 1.0 (SE) yr, body fat 11.9 +/- 1.9%, body mass index 26.1 +/- 1.1 kg/m2; n = 10] men before and after 3 h of hyperinsulinemia (hyperinsulinemic-euglycemic clamp). Muscle was analyzed for mRNA of type I, IIa, and IIx MHC isoforms. Hyperinsulinemia (mean of 1,065.7 +/- 9.8 pmol/l during minutes 20 to 180) did not change (P > 0.05) the mRNA concentration of either the type I MHC or type IIA MHC isoforms. In contrast, type IIX MHC mRNA increased (P < 0.05) with hyperinsulinemia compared with the fasted condition. These data indicate that hyperinsulinemia rapidly increases type IIx MHC mRNA in human skeletal muscle. PMID- 10368346 TI - Ultrastructural muscle damage in young vs. older men after high-volume, heavy resistance strength training. AB - This study assessed ultrastructural muscle damage in young (20-30 yr old) vs. older (65-75 yr old) men after heavy-resistance strength training (HRST). Seven young and eight older subjects completed 9 wk of unilateral leg extension HRST. Five sets of 5-20 repetitions were performed 3 days/wk with variable resistance designed to subject the muscle to near-maximal loads during every repetition. Biopsies were taken from the vastus lateralis of both legs, and muscle damage was quantified via electron microscopy. Training resulted in a 27% strength increase in both groups (P < 0.05). In biopsies before training in the trained leg and in all biopsies from untrained leg, 0-3% of muscle fibers exhibited muscle damage in both groups (P = not significant). After HRST, 7 and 6% of fibers in the trained leg exhibited damage in the young and older men, respectively (P < 0.05, no significant group differences). Myofibrillar damage was primarily focal, confined to one to two sarcomeres. Young and older men appear to exhibit similar levels of muscle damage at baseline and after chronic HRST. PMID- 10368347 TI - Reversal of weightlessness-induced musculoskeletal losses with androgens: quantification by MRI. AB - Microgravity causes rapid decrement in musculoskeletal mass is associated with a marked decrease in circulatory testosterone levels, as we reported in hindlimb suspended (HLS) rats. In this model which simulates microgravity, we hypothesized that testosterone supplementation should prevent these losses, and we tested this in two studies. Muscle volumes and bone masses were quantitated by using magnetic resonance imaging (MRI) on day 12. In the first study, 12-wk-old Sprague-Dawley rats that were HLS for 12 days lost 28.5% of muscle volume (53.3 +/- 4.8 vs. 74.5 +/- 3.6 cm3 in the ground control rats; P < 0.001) and had a 5% decrease in bone mineral density (BMD) (P < 0.05). In the second study, 30 male 12-wk-old Wistar rats were HLS and were administered either a vehicle (control), testosterone, or nandrolone decanoate (ND). An additional 20 rats were used as ground controls, one-half of which received testosterone. HLS rats had a significant reduction in muscle volume (42.9 +/- 3.0 vs. 56 +/- 1.8 cm3 in ground control rats; P < 0.01). Both testosterone and ND treatments prevented this muscle loss (51.5 +/- 2 and 51.6 +/- 1.2 cm3, respectively; a 63% improvement; P < 0. 05). There were no statistical differences between the two active treatment groups nor with the ground controls. Similarly, there was an 85% improvement in BMD in the testosterone group (1.15 +/- 0.04 vs. 1.04 +/- 0.04 density units in vehicle controls; P < 0.05) and a 76% improvement in the ND group (1.13 +/- 0.07 density units), whereas ground control rats had a BMD of 1.17 +/- 0.03 density units. Because serum testosterone levels are markedly reduced in this model of simulated microgravity, androgen replacement seems to be a rational countermeasure to prevent microgravity-induced musculoskeletal losses. PMID- 10368349 TI - Acute manipulations of plasma volume alter arterial pressure responses during Valsalva maneuvers. AB - The effects of changes in blood volume on arterial pressure patterns during the Valsalva maneuver are incompletely understood. In the present study we measured beat-to-beat arterial pressure and heart rate responses to supine Valsalva maneuvers during normovolemia, hypovolemia induced with intravenous furosemide, and hypervolemia induced with ingestion of isotonic saline. Valsalva responses were analyzed according to the four phases as previously described (W. F. Hamilton, R. A. Woodbury, and H. T. Harper, Jr. JAMA 107: 853-856, 1936; W. F. Hamilton, R. A. Woodbury, and H. T. Harper, Jr. Am. J. Physiol. 141: 42-50, 1944). Phase I is the initial onset of straining, which elicits a rise in arterial pressure; phase II is the period of straining, during which venous return is impeded and pressure falls (early) and then partially recovers (late); phase III is the initial release of straining; and phase IV consists of a rapid "overshoot" of arterial pressure after the release. During hypervolemia, early phase II arterial pressure decreases were significantly less than those during hypovolemia, thus making the response more "square." Systolic pressure hypervolemic vs. hypovolemic falls were -7.4 +/- 2.1 vs. -30.7 +/- 7 mmHg (P = 0.005). Diastolic pressure hypervolemic vs. hypovolemic falls were -2.4 +/- 1.6 vs. -15.2 +/- 2.6 mmHg (P = 0.05). A significant direct correlation was found between plasma volume and phase II systolic pressure falls, and a significant inverse correlation was found between plasma volume and phase III-IV systolic pressure overshoots. Heart rate responses to systolic pressure falls during phase II were significantly less during hypovolemia than during hypervolemia (0.7 +/- 0.2 vs. 2.82 +/- 0.2 beats. min-1. mmHg-1; P = 0.05) but were not different during phase III-IV overshoots. We conclude that acute changes in intravascular volume from hypovolemia to hypervolemia affect cardiovascular responses, particularly arterial pressure changes, to the Valsalva maneuver and should be considered in both clinical and research applications of this maneuver. PMID- 10368348 TI - Sodium-free fluid ingestion decreases plasma sodium during exercise in the heat. AB - This study assessed whether replacing sweat losses with sodium-free fluid can lower the plasma sodium concentration and thereby precipitate the development of hyponatremia. Ten male endurance athletes participated in one 1-h exercise pretrial to estimate fluid needs and two 3-h experimental trials on a cycle ergometer at 55% of maximum O2 consumption at 34 degrees C and 65% relative humidity. In the experimental trials, fluid loss was replaced by distilled water (W) or a sodium-containing (18 mmol/l) sports drink, Gatorade (G). Six subjects did not complete 3 h in trial W, and four did not complete 3 h in trial G. The rate of change in plasma sodium concentration in all subjects, regardless of exercise time completed, was greater with W than with G (-2.48 +/- 2.25 vs. -0.86 +/- 1.61 mmol. l-1. h-1, P = 0.0198). One subject developed hyponatremia (plasma sodium 128 mmol/l) at exhaustion (2.5 h) in the W trial. A decrease in sodium concentration was correlated with decreased exercise time (R = 0.674; P = 0.022). A lower rate of urine production correlated with a greater rate of sodium decrease (R = -0. 478; P = 0.0447). Sweat production was not significantly correlated with plasma sodium reduction. The results show that decreased plasma sodium concentration can result from replacement of sweat losses with plain W, when sweat losses are large, and can precipitate the development of hyponatremia, particularly in individuals who have a decreased urine production during exercise. Exercise performance is also reduced with a decrease in plasma sodium concentration. We, therefore, recommend consumption of a sodium-containing beverage to compensate for large sweat losses incurred during exercise. PMID- 10368350 TI - Human skeletal sarcoplasmic reticulum Ca2+ uptake and muscle function with aging and strength training. AB - This study investigated the adaptations of skeletal muscle sarcoplasmic reticulum (SR) Ca2+ uptake, relaxation, and fiber types in young (YW) and elderly women (EW) to high-resistance training. Seventeen YW (18-32 yr) and 11 EW (64-79 yr) were assessed for 1) electrically evoked relaxation time and rate of the quadriceps femoris; and 2) maximal rates of SR Ca2+ uptake and Ca2+-ATPase activity and relative fiber-type areas, analyzed from muscle biopsies of the vastus lateralis. EW had significantly slower relaxation rates and times, decreased SR Ca2+ uptake and Ca2+-ATPase activity, and a larger relative type I fiber area than did YW. A subgroup of 9 young (YWT) and 10 elderly women (EWT) performed 12 wk of high-resistance training (8 repetition maximum) of the quadriceps and underwent identical testing procedures pre- and posttraining. EWT significantly increased their SR Ca2+ uptake and Ca2+-ATPase activity in response to training but showed no alterations in speed of relaxation or relative fiber type areas. In YWT none of the variables was altered after resistance training. These findings suggest that 1) a reduced SR Ca2+ uptake in skeletal muscle of elderly women was partially reversed with resistance training and 2) SR Ca2+ uptake in the vastus lateralis was not the rate-limiting mechanism for the slowing of relaxation measured from electrically evoked quadriceps muscle of elderly women. PMID- 10368351 TI - Detection of changes in lung tissue properties with multiple-indicator dilution. AB - We evaluated the potential utility of a group of indicators, each of which targets a particular tissue property, as indicators in the multiple-indicator dilution method to detect and to identify abnormalities in lung tissue properties resulting from lung injury models. We measured the pulmonary venous outflow concentration vs. time curves of [14C]diazepam, 3HOH, [14C]phenylethylamine, and a vascular reference indicator following their bolus injection into the pulmonary artery of isolated perfused rabbit lungs under different experimental conditions, resulting in changes in the lung tissue composition. The conditions included granulomatous inflammation, induced by the intravenous injection of complete Freund's adjuvant (CFA), and intratracheal fluid instillation, each of which resulted in similar increases in lung wet weight. Each of these conditions resulted in a unique pattern among the concentration vs. time outflow curves of the indicators studied. The patterns were quantified by using mathematical models describing the pulmonary disposition of each of the indicators studied. A unique model parameter vector was obtained for each condition, demonstrating the ability to detect and to identify changes in lung tissue properties by using the appropriate group of indicators in the multiple-indicator dilution method. One change that was particularly interesting was a CFA-induced change in the disposition of diazepam, suggestive of a substantial increase in peripheral-type benzodiazepine receptors in the inflamed lungs. PMID- 10368352 TI - Pattern of expiratory muscle activation during lower thoracic spinal cord stimulation. AB - Large positive airway pressures (Paws) can be generated by lower thoracic spinal cord stimulation (SCS), which may be a useful method of restoring cough in spinal cord-injured patients. Optimal electrode placement, however, requires an assessment of the pattern of current spread during SCS. Studies were performed in anesthetized dogs to assess the pattern of expiratory muscle recruitment during SCS applied at different spinal cord levels. A multicontact stimulating electrode was positioned over the surface of the lower thoracic and upper lumbar spinal cord. Recording electromyographic electrodes were placed at several locations in the abdominal and internal intercostal muscles. SCS was applied at each lead, in separate trials, with single shocks of 0.2-ms duration. The intensity of stimulation was adjusted to determine the threshold for development of the compound action potential at each electrode lead. The values of current threshold for activation of each muscle formed parabolas with minimum values at specific spinal root levels. The slopes of the parabolas were relatively steep, indicating that the threshold for muscle activation increases rapidly at more cephalad and caudal sites. These results were compared with the effectiveness of SCS (50 Hz; train duration, 1-2 s) at different spinal cord levels to produce changes in Paw. Stimulation at the T9 and T10 spinal cord level resulted in the largest positive Paws with a single lead. At these sites, threshold values for activation of the internal intercostal (7-11th interspaces) upper portions of external oblique, rectus abdominis, and transversus abdominis were near their minimum. Threshold values for activation of the caudal portions of the abdominal muscles were high (>50 mA). Our results indicate that 1) activation of the more cephalad portions of the abdominal muscles is more important than activation of caudal regions in the generation of positive Paws and 2) it is not possible to achieve complete activation of the expiratory muscles with a single electrode lead by using modest current levels. In support of this latter conclusion, a two-electrode lead system results in more uniform expiratory muscle activation and significantly greater changes in Paw. PMID- 10368353 TI - Effects of vagotomy on cardiovascular response to periodic apneas in sedated pigs. AB - There are few studies investigating the influence of vagally mediated reflexes on the cardiovascular response to apneas. In 12 sedated preinstrumented pigs, we studied the effects of vagotomy during apneas, controlling for apnea periodicity and thoracic mechanical effects. Nonobstructive apneas were produced by paralyzing and mechanically ventilating the animals, then turning the ventilator off and on every 30 s. Before vagotomy, relative to baseline, apnea caused increased mean arterial pressure (MAP; +19 +/- 25%, P < 0.05), systemic vascular resistance (SVR; +33 +/- 16%, P < 0.0005), and heart rate (HR; +5 +/- 6%, P < 0.05) and decreased cardiac output (CO) and stroke volume (SV; -16 +/- 10% P < 0.001). After vagotomy, no significant change occurred in MAP, SVR, and SV during apneas, but CO and HR increased relative to baseline. HR was always greater ( approximately 14%, P < 0.01) during the interapneic interval compared with during apnea. We conclude that vagally mediated reflexes are important mediators of the apneic pressor response. HR increases after apnea termination are related, at least in part, to nonvagally mediated reflexes. PMID- 10368355 TI - Effects of resistance training on selected indexes of immune function in elderly women. AB - Women aged 67-84 yr were randomly assigned to either resistance exercise (RE, n = 15) or control group (C, n = 14). RE group completed 10 wk of resistance training, whereas C group maintained normal activity. Blood samples were obtained from the RE group (at the same time points as for resting C) at rest, immediately after resistance exercise, and 2 h after exercise before (week 0) and after (week 10) training. Mononuclear cell (CD3+, CD3+CD4+, CD3+CD8+, CD19+, and CD3 CD16+CD56+) number, lymphocyte proliferative (LP) response to mitogen, natural cell-mediated cytotoxicity (NCMC), and serum cortisol levels were determined. Strength increased significantly in RE subjects (%change 8-repetition maximum = 148%). No significant group, exercise time, or training effects were found for CD3+, CD3+CD4+, or CD3+CD8+ cells, but there was a significant exercise time effect for CD3-CD16+CD56+ cells. LP response was not different between groups, across exercise time, or after training. NCMC was increased immediately after exercise for RE subjects at week 0 and for RE and C groups at week 10. The week 0 and week 10 NCMC values were above baseline for both RE and C groups 2 h after exercise. In conclusion, acute resistance exercise did not result in postexercise suppression of NCMC or LP, and 10 wk of resistance training did not influence resting immune measures in women aged 67-84 yr. PMID- 10368354 TI - Ventilatory effects of 8 h of isocapnic hypoxia with and without beta-blockade in humans. AB - This study investigated whether changing sympathetic activity, acting via beta receptors, might induce the progressive ventilatory changes observed in response to prolonged hypoxia. The responses of 10 human subjects to four 8-h protocols were compared: 1) isocapnic hypoxia (end-tidal PO2 = 50 Torr) plus 80-mg doses of oral propranolol; 2) isocapnic hypoxia, as in protocol 1, with oral placebo; 3) air breathing with propranolol; and 4) air breathing with placebo. Exposures were conducted in a chamber designed to maintain end-tidal gases constant by computer control. Ventilation (VE) was measured at regular intervals throughout. Additionally, the subjects' ventilatory hypoxic sensitivity and their residual VE during hyperoxia (5 min) were assessed at 0, 4, and 8 h by using a dynamic end tidal forcing technique. beta-Blockade did not significantly alter either the rise in VE seen during 8 h of isocapnic hypoxia or the changes observed in the acute hypoxic ventilatory response and residual VE in hyperoxia over that period. The results do not provide evidence that changes in sympathetic activity acting via beta-receptors play a role in the mediation of ventilatory changes observed during 8 h of isocapnic hypoxia. PMID- 10368356 TI - Acute hypoxic pulmonary vasoconstriction in conscious dogs decreases renin and is unaffected by losartan. AB - Acute hypoxic pulmonary vasoconstriction (HPV) may be mediated by vasoactive peptides. We studied eight conscious, chronically tracheostomized dogs kept on a standardized dietary sodium intake. Normoxia (40 min) was followed by hypoxia (40 min, breathing 10% oxygen, arterial oxygen pressures 36 +/- 1 Torr) during both control (Con) and losartan experiments (Los; iv infusion of 100 microg. min-1. kg 1 losartan). During hypoxia, minute ventilation (by 0.9 l/min in Con, by 1.3 l/min in Los), cardiac output (by 0.36 l/min in Con, by 0.30 l/min in Los), heart rate (by 11 beats/min in Con, by 30 beats/min in Los), pulmonary artery pressure (by 9 mmHg in both protocols), and pulmonary vascular resistance (by 280 and 254 dyn. s. cm-5 in Con and Los, respectively) increased. Mean arterial pressure and systemic vascular resistance did not change. In Con, PRA decreased from 4.2 +/- 0.7 to 2.5 +/- 0.5 ng ANG I. ml-1. h-1, and plasma ANG II decreased from 11.9 +/- 3.0 to 8.2 +/- 2.1 pg/ml. The renin-angiotensin system is inhibited during acute hypoxia despite sympathetic activation. Under these conditions, ANG II AT1 receptor antagonism does not attenuate HPV. PMID- 10368357 TI - Predicting risk of decompression sickness in humans from outcomes in sheep. AB - In animals, the response to decompression scales as a power of species body mass. Consequently, decompression sickness (DCS) risk in humans should be well predicted from an animal model with a body mass comparable to humans. No-stop decompression outcomes in compressed air and nitrogen-oxygen dives with sheep (n = 394 dives, 14.5% DCS) and humans (n = 463 dives, 4.5% DCS) were used with linear-exponential, probabilistic modeling to test this hypothesis. Scaling the response parameters of this model between species (without accounting for body mass), while estimating tissue-compartment kinetic parameters from combined human and sheep data, predicts combined risk better, based on log likelihood, than do separate sheep and human models, a combined model without scaling, and a kinetic scaled model. These findings provide a practical tool for estimating DCS risk in humans from outcomes in sheep, especially in decompression profiles too risky to test with humans. This model supports the hypothesis that species of similar body mass have similar DCS risk. PMID- 10368358 TI - Comparison of short-term diet and exercise on insulin action in individuals with abnormal glucose tolerance. AB - The effects of a 10-day low-calorie diet (LCD; n = 8) or exercise training (ET; n = 8) on insulin secretion and action were compared in obese men (n = 9) and women (n = 7), aged 53 +/- 1 yr, with abnormal glucose tolerance by using a hyperglycemic clamp with superimposed arginine infusion and a high-fat drink. Body mass (LCD, 115 +/- 5 vs. 110 +/- 5 kg; ET, 111 +/- 7 vs. 109 +/- 7 kg; P < 0. 01) and fasting plasma glucose (LCD, 115 +/- 10 vs. 99 +/- 4 mg/dl; ET, 112 +/ 4 vs. 101 +/- 5 mg/dl, P < 0.01) and insulin (LCD, 23.9 +/- 5.6 vs. 15.2 +/- 3.9 microU/ml; ET, 17.6 +/- 1.9 vs. 13.9 +/- 2. 4 microU/ml; P < 0.05) decreased in both groups. There was a 40% reduction in plasma insulin during hyperglycemia (0 45 min) after LCD (peak: 118 +/- 18 vs. 71 +/- 14 microU/ml; P < 0.05) and ET (69 +/- 14 vs. 41 +/- 7 microU/ml; P < 0.05) and trends for reductions during arginine infusion and a high-fat drink. The 56% increase in glucose uptake after ET (4.95 +/- 0.90 vs. 7.74 +/- 0.82 mg. min-1. kg fat-free mass-1; P < 0.01) was significantly (P < 0.01) greater than the 19% increase (5.72 +/- 1.12 vs. 6.80 +/ 0.94 mg. min-1. kg fat-free mass-1; P = not significant) that occurred after LCD. The marked increase in glucose disposal after ET, despite lower insulin levels, suggests that short-term exercise is more effective than diet in enhancing insulin action in individuals with abnormal glucose tolerance. PMID- 10368360 TI - Nitric oxide does not modulate whole body oxygen consumption in anesthetized dogs. AB - The effects of the NO synthase inhibitor NG-nitro-L-arginine methyl ester (L NAME) and the NO donor sodium nitroprusside (SNP) on whole body O2 consumption (VO2) were assessed in 16 dogs anesthetized with fentanyl or isoflurane. Cardiac output (CO) and mean arterial pressure (MAP) were measured with standard methods and were used to calculate VO2 and systemic vascular resistance (SVR). Data were obtained in each dog under the following conditions: 1) Control 1, 2) SNP (30 microg. kg-1. min-1 iv) 3) Control 2, 4) L-NAME (10 mg/kg iv), and 5) SNP and adenosine (30 and 600 microg. kg-1. min-1 iv, respectively) after L-NAME. SNP reduced MAP by 29 +/- 3% and SVR by 47 +/- 3%, while it increased CO by 39 +/- 9%. L-NAME had opposite effects; it increased MAP and SVR by 24 +/- 4% and 103 +/ 11%, respectively, and it decreased CO by 37 +/- 3%. Neither agent changed VO2 from the baseline value of 4.3 +/- 0.2 ml. min-1. kg-1, since the changes in CO were offset by changes in the arteriovenous O2 difference. Both SNP and adenosine returned CO to pre-L-NAME values, but VO2 was unaffected. We conclude that 1) basally released endogenous NO had a tonic systemic vasodilator effect, but it had no influence on VO2; 2) SNP did not alter VO2 before or after inhibition of endogenous NO production; 3) the inability of L-NAME to increase VO2 was not because CO, i.e., O2 supply, was reduced below the critical level. PMID- 10368359 TI - Age and renal prostaglandin inhibition during exercise and heat stress. AB - Aging is associated with a number of physiological changes that may cause the kidney to rely to a greater extent on vasodilatory PGs for normal functioning. Acute exercise has been shown to cause renal vasoconstriction that may be partially buffered by vasodilatory PGs. To determine the relative importance of renal PGs during exercise in older adults, we compared the renal effects of the PG inhibitor ibuprofen (1.2 g/day for 3 days) vs. a placebo control in a cohort of eight younger (24 +/- 2 yr) and eight older (64 +/- 2 yr) women during treadmill exercise ( approximately 57% maximal oxygen consumption) in the heat (36 degrees C). This over-the-counter dose of ibuprofen reduced renal PG (i.e., PGE2) excretion by 47% (P < 0. 05). Acute exercise in the heat caused dramatic decreases in glomerular filtration rate, renal blood flow, and sodium excretion in both age groups. PG inhibition was associated with greater decreases in urine production and free water clearance (P < 0.05). There were no drug-related declines in glomerular filtration rate or renal blood flow. We conclude that PG inhibition has only modest effects on renal function during exercise. Also, the lack of hemodynamic changes with PG inhibition indicates that healthy well hydrated older women are not in a renal PG-dependent state. PMID- 10368362 TI - Alterations in pulmonary surfactant after rapid arousal from torpor in the marsupial Sminthopsis crassicaudata. AB - Torpor in the dunnart, Sminthopsis crassicaudata, alters surfactant lipid composition and surface activity. Here we investigated changes in surfactant composition and surface activity over 1 h after rapid arousal from torpor (15-30 degrees C at 1 degrees C/min). We measured total phospholipid (PL), disaturated PL (DSP), and cholesterol (Chol) content of surfactant lavage and surface activity (measured at both 15 and 37 degrees C in the captive bubble surfactometer). Immediately after arousal, Chol decreased (from 4.1 +/- 0.05 to 2.8 +/- 0.3 mg/g dry lung) and reached warm-active levels by 60 min after arousal. The Chol/DSP and Chol/PL ratios both decreased to warm-active levels 5 min after arousal because PL, DSP, and the DSP/PL ratio remained elevated over the 60 min after arousal. Minimal surface tension and film compressibility at 17 mN/m at 37 degrees C both decreased 5 min after arousal, correlating with rapid changes in surfactant Chol. Therefore, changes in lipids matched changes in surface activity during the postarousal period. PMID- 10368361 TI - Estrogen has rapid tissue-specific effects on rat bone. AB - The decrease in cancellous bone formation after estrogen treatment is generally thought to be coupled with a prior decrease in bone resorption. To test the possibility that estrogen has rapid tissue-specific actions on bone metabolism, we determined the time course (1-32 h) effects of diethylstilbestrol on steady state mRNA levels for immediate-response genes, extracellular matrix proteins, and signaling peptides in the proximal tibial metaphysis and uterus by using Northern blot and RNase protection assays. The regulation of signaling peptides by estrogen, although tissue specific, followed a similar time course in bone and uterus. The observed rapid decreases in expression of insulin-like growth factor I, a growth factor associated with bone formation; decreases in mRNA levels for bone matrix proteins; evidence for reduced bone matrix synthesis; failure to detect rapid increases in mRNA levels for signaling peptides implicated in mediating the inhibitory effects of estrogen on bone resorption (interleukin-1 and -6) as well as other cytokines that can increase bone resorption; and the comparatively long duration of the bone remodeling cycle in rats indicate that estrogen can decrease bone formation by a mechanism that does not require a prior reduction in bone resorption. PMID- 10368363 TI - Sympathetic responses to head-down rotations in humans. AB - Muscle sympathetic nerve activity (MSNA) increases with head-down neck flexion (HDNF). The present study had three aims: 1) to examine sympathetic and vascular responses to two different magnitudes of HDNF; 2) to examine these same responses during prolonged HDNF; and 3) to determine the influence of nonspecific pressure receptors in the head on MSNA. The first experiment tested responses to two static head positions in the vertical axis [HDNF and intermediate HDNF (I-HDNF; approximately 50% of HDNF)]. MSNA increased above baseline during both I-HDNF and HDNF (from 219 +/- 36 to 301 +/- 47 and from 238 +/- 42 to 356 +/- 59 units/min, respectively; P < 0.01). Calf blood flow (CBF) decreased and calf vascular resistance increased during both I-HDNF and HDNF (P < 0.01). Both the increase in MSNA and the decrease in CBF were linearly related to the magnitude of the downward head rotations (P < 0.01). The second experiment tested responses during prolonged HDNF. MSNA increased (from 223 +/- 63 to 315 +/- 79 units/min; P < 0.01) and CBF decreased (from 3.2 +/- 0.4 to 2.6 +/- 0.04 ml. 100 ml-1. min-1; P < 0.01) at the onset of HDNF. These responses were maintained throughout the 30 min period. Mean arterial blood pressure gradually increased during the 30 min of HDNF (from 94 +/- 4 to 105 +/- 3 mmHg; P < 0.01). In a third experiment, head down neck extension was performed with subjects in the supine position. Unlike HDNF, head-down neck extension did not affect MSNA. The results from these studies demonstrate that MSNA: 1) increases in magnitude as the degree of HDNF increases; 2) remains elevated above baseline during prolonged HDNF; and 3) responses during HDNF are not associated with nonspecific receptors in the head activated by increases in cerebral pressure. PMID- 10368364 TI - A physiological model for predicting carboxyhemoglobin formation from exposure to carbon monoxide in rats. AB - A time-dependent simulation model, based on the Coburn-Forster-Kane equation, was written in Advanced Continuous Simulation Language to predict carboxyhemoglobin (HbCO) formation and dissociation in F-344 rats during and after exposure to 500 parts/million CO for 1 h. Blood-gas analysis and CO-oximetry were performed on samples collected during exposure and off-gassing of CO. Volume displacement plethysmography was used to measure minute ventilation (VE) during exposure. CO diffusing capacity in the lung (DLCO) was also measured. Other model parameters measured in the animals included blood pH, total blood volume, and Hb concentration. Comparisons between model predictions using values for VE, DLCO, and the Haldane coefficient cited in the literature and predictions using measured VE, DLCO, and calculated Haldane coefficient for individual animals were made. General model predictions using values for model parameters derived from the literature agreed with published HbCO values by a factor of 0.987 but failed to simulate experimental data. On average, the general model overpredicted measured HbCO level by nearly 9%. A specific model using the means of measured variables predicted HbCO concentration within a factor of 0.993. When experimentally observed parameter fluctuations were included, the specific model predictions reflected experimental effects on HbCO formation. PMID- 10368365 TI - Longitudinal distribution of chlorine absorption in human airways: comparison of nasal and oral quiet breathing. AB - The fraction of an inspired chlorine (Cl2) bolus absorbed during a single breath (Lambda) was measured as a function of bolus penetration (VP) into the respiratory system of five male and five female nonsmokers during both nasal and oral breathing at a quiet respiratory flow of 250 ml/s. The correspondence between VP and specific anatomic landmarks was found for each subject by a combination of acoustic reflection and nitrogen washout measurements. For both nasal and oral breathing, Lambda reached approximately 0. 95 at the distal end of the upper airways and reached 1.00 within the lower conducting airways. The values of a regional mass transfer parameter computed from the Lambda-VP data indicated that the resistance to Cl2 diffusion in the airway mucosa was negligible compared with the diffusion resistance in the respired gas. Changing the peak inhaled Cl2 concentration from 0.5 to 3.0 parts/million did not significantly affect the distribution of Cl2 absorption, suggesting that the underlying mass transport and chemical reaction processes were linear with respect to Cl2 concentration. PMID- 10368366 TI - Posture effects on timing of abdominal muscle activity during stimulated ventilation. AB - In humans during stimulated ventilation, substantial abdominal muscle activity extends into the following inspiration as postexpiratory expiratory activity (PEEA) and commences again during late inspiration as preexpiratory expiratory activity (PREA). We hypothesized that the timing of PEEA and PREA would be changed systematically by posture. Fine-wire electrodes were inserted into the rectus abdominis, external oblique, internal oblique, and transversus abdominis in nine awake subjects. Airflow, end-tidal CO2, and moving average electromyogram (EMG) signals were recorded during resting and CO2-stimulated ventilation in both supine and standing postures. Phasic expiratory EMG activity (tidal EMG) of the four abdominal muscles at any level of CO2 stimulation was greater while standing. Abdominal muscle activities during inspiration, PEEA, and PREA, were observed with CO2 stimulation, both supine and standing. Change in posture had a significant effect on intrabreath timing of expiratory muscle activation at any level of CO2 stimulation. The transversus abdominis showed a significant increase in PEEA and a significant decrease in PREA while subjects were standing; similar changes were seen in the internal oblique. We conclude that changes in posture are associated with significant changes in phasic expiratory activity of the four abdominal muscles, with systematic changes in the timing of abdominal muscle activity during early and late inspiration. PMID- 10368367 TI - Airway remodeling in asthma amplifies heterogeneities in smooth muscle shortening causing hyperresponsiveness. AB - Although airway remodeling and inflammation in asthma can amplify the constriction response of a single airway, their influence on the structural changes in the whole airway network is unknown. We present a morphometric model of the human lung that incorporates cross-sectional wall areas corresponding to the adventitia, airway smooth muscle (ASM), and mucosa for healthy and mildly and severely asthmatic airways and the influence of parenchymal tethering. A heterogeneous ASM percent shortening stimulus is imposed, causing distinct constriction patterns for healthy and asthmatic airways. We calculate lung resistance and elastance from 0.1 to 5 Hz. We show that, for a given ASM stimulus, the distribution of wall area in asthmatic subjects will amplify not only the mean but the heterogeneity of constriction in the lung periphery. Moreover, heterogeneous ASM shortening that would produce only mild changes in the healthy lung can cause hyperresponsive changes in lung resistance and elastance at typical breathing rates in the asthmatic lung, even with relatively small increases in airway resistance. This condition arises when airway closures occur randomly in the lung periphery. We suggest that heterogeneity is a crucial determinant of hyperresponsiveness in asthma and that acute asthma is more a consequence of extensive airway wall inflammation and remodeling, predisposing the lung to produce an acute pattern of heterogeneous constriction. PMID- 10368368 TI - Skeletal muscle phosphocreatine recovery in exercise-trained humans is dependent on O2 availability. AB - In skeletal muscle, phosphocreatine (PCr) recovery from submaximal exercise has become a reliable and accepted measure of muscle oxidative capacity. During exercise, O2 availability plays a role in determining maximal oxidative metabolism, but the relationship between O2 availability and oxidative metabolism measured by 31P-magnetic resonance spectroscopy (MRS) during recovery from exercise has never been studied. We used 31P-MRS to study exercising human gastrocnemius muscle under conditions of varied fractions of inspired O2 (FIO2) to test the hypothesis that varied O2 availability modulates PCr recovery from submaximal exercise. Six male subjects performed three bouts of 5-min steady state submaximal plantar flexion exercise followed by 5 min of recovery in a 1.5 T magnet while breathing three different FIO2 concentrations (0.10, 0. 21, and 1.00). Under each FIO2 treatment, the PCr recovery time constants were significantly different, being longer in hypoxia [33. 5 +/- 4.1 s (SE)] and shorter in hyperoxia (20.0 +/- 1.8 s) than in normoxia (25.0 +/- 2.7 s) (P 300 s-1 in 35 KP but then declined to <1 s-1 by 90 min. The decrease in frequency was not accompanied by a decrease in MEPC average amplitude. Nerve terminals accumulated the activity dependent dye FM1-43 when exposed to the dye for the final 6 min of a 120-min exposure to 35 KP. Thus synaptic membrane endocytosis continued at a high rate, although MEPCs occurred infrequently. After a 120-min exposure in 35 KP, nerve terminals accumulated FM1-43 and then destained, confirming that exocytosis also still occurred at a high rate. These results demonstrate that recycled cholinergic synaptic vesicles that were not refilled with ACh continued to dock and undergo exocytosis after membrane retrieval. Thus transport of ACh into recycled cholinergic vesicles is not a requirement for repeated cycles of exocytosis and retrieval of synaptic vesicle membrane during prolonged stimulation of motor nerve terminals. PMID- 10368391 TI - Surround suppression in the responses of primate SA1 and RA mechanoreceptive afferents mapped with a probe array. AB - Twenty-four slowly adapting type 1 (SA1) and 26 rapidly adapting (RA) cutaneous mechanoreceptive afferents in the rhesus monkey were studied with an array of independently controlled, punctate probes that covered an entire fingerpad. Each afferent had a receptive field (RF) on a single fingerpad and was studied at 73 skin sites (50 mm2). The entire array was lowered to 1.6 to 3.0 mm below the point of initial skin contact (the background indentation) before delivering indentations with one to seven probes. Indentations were generally limited to 100 microm to minimize gross mechanical interactions. There were two major, new findings. 1) The discharge rates of both SA1 and RA afferents were strongly affected by the number of probes indenting the RF simultaneously. The effect was exponential. Each increase in probe number reduced the response by 24% in SA1 and 12% in RA afferents on average. When seven probes indented the skin simultaneously, the impulse rates in SA1 and RA afferents were reduced to 20 and 40% of the rates evoked by a single probe at the hot spot (all indentations were 100 microm). This shows that before any synaptic interaction in the CNS there is already a mechanism analogous to surround inhibition that suppresses an afferent's responses to uniform indentation and makes it especially sensitive to deviations from spatial uniformity. 2) The responses of both SA1 and RA afferents were independent of background array depth over the range from 1.6 to 3 mm below the point of initial skin contact. This shows that the neural responses to elements raised above a background are independent of the applied force over a wide range of forces. To relate the background depths to indentation force and to compare humans and monkeys, we studied the biomechanics of indentation with a uniform surface. A remarkable result is that the force-displacement relationships in humans and monkeys were the same; the skin is highly compliant for the first 2 3 mm of indentation and then becomes much stiffer. The results were the same in alert humans and monkeys and in monkeys anesthetized with pentobarbital. Ketamine anesthesia made the skin much stiffer and reduced the compliant range substantially. PMID- 10368392 TI - Influence of head position on the spatial representation of acoustic targets. AB - Sound localization in humans relies on binaural differences (azimuth cues) and monaural spectral shape information (elevation cues) and is therefore the result of a neural computational process. Despite the fact that these acoustic cues are referenced with respect to the head, accurate eye movements can be generated to sounds in complete darkness. This ability necessitates the use of eye position information. So far, however, sound localization has been investigated mainly with a fixed head position, usually straight ahead. Yet the auditory system may rely on head motor information to maintain a stable and spatially accurate representation of acoustic targets in the presence of head movements. We therefore studied the influence of changes in eye-head position on auditory guided orienting behavior of human subjects. In the first experiment, we used a visual-auditory double-step paradigm. Subjects made saccadic gaze shifts in total darkness toward brief broadband sounds presented before an intervening eye-head movement that was evoked by an earlier visual target. The data show that the preceding displacements of both eye and head are fully accounted for, resulting in spatially accurate responses. This suggests that auditory target information may be transformed into a spatial (or body-centered) frame of reference. To further investigate this possibility, we exploited the unique property of the auditory system that sound elevation is extracted independently from pinna related spectral cues. In the absence of such cues, accurate elevation detection is not possible, even when head movements are made. This is shown in a second experiment where pure tones were localized at a fixed elevation that depended on the tone frequency rather than on the actual target elevation, both under head fixed and -free conditions. To test, in a third experiment, whether the perceived elevation of tones relies on a head- or space-fixed target representation, eye movements were elicited toward pure tones while subjects kept their head in different vertical positions. It appeared that each tone was localized at a fixed, frequency-dependent elevation in space that shifted to a limited extent with changes in head elevation. Hence information about head position is used under static conditions too. Interestingly, the influence of head position also depended on the tone frequency. Thus tone-evoked ocular saccades typically showed a partial compensation for changes in static head position, whereas noise-evoked eye-head saccades fully compensated for intervening changes in eye-head position. We propose that the auditory localization system combines the acoustic input with head-position information to encode targets in a spatial (or body-centered) frame of reference. In this way, accurate orienting responses may be programmed despite intervening eye-head movements. A conceptual model, based on the tonotopic organization of the auditory system, is presented that may account for our findings. PMID- 10368393 TI - Electrical stimulation of the horizontal limb of the diagonal band of broca modulates population EPSPs in piriform cortex. AB - Electrical stimulation of the horizontal limb of the diagonal band of Broca (HDB) was coupled with recording of evoked potentials in the piriform cortex. Stimulation of the HDB caused an enhancement of the late, disynaptic component of the evoked potential elicited by stimulation of the lateral olfactory tract but caused a suppression of the synaptic potential elicited by stimulation of the posterior piriform cortex. The muscarinic antagonist scopolamine blocked both effects of HDB stimulation. The enhancement of disynaptic potentials could be due to cholinergic depolarization of pyramidal cells, whereas the suppression of potentials evoked by posterior piriform stimulation could be due to presynaptic inhibition of intrinsic fiber synaptic transmission by acetylcholine. PMID- 10368394 TI - Serotonergic modulation of synapses in the developing gerbil lateral superior olive. AB - The lateral superior olive (LSO) is a primary site of binaural convergence that responds selectively to changes in interaural level difference (ILD) by integrating ipsilateral excitatory and contralateral inhibitory inputs. The circuit matures during the first three postnatal weeks, undergoing several structural and functional changes that are influenced by afferent activity. Therefore modulation of synaptic activity by neuromodulators may participate in the maturation of this circuit. The present study describes robust effects of serotonin (5-HT) on LSO synaptic function. Using whole cell voltage-clamp recording from gerbil LSO neurons (postnatal days 6-13) in an in vitro slice preparation, we have identified several distinct forms of serotonergic modulation of spontaneous and evoked synaptic transmission. First, 1-2 min application of 5 HT (100 microM) activated prolonged bursts of spontaneous inhibitory postsynaptic currents (IPSCs). However, there was an age-dependent decline, such that this effect rarely was observed beyond postnatal day 8. 5-HT apparently increased the excitability of inhibitory afferents, because 5-HT-induced IPSCs were blocked by tetrodotoxin. A second effect of 5-HT was to depress rapidly and profoundly the amplitude of electrically evoked excitatory postsynaptic currents (EPSCs). In contrast, 5-HT also depressed evoked IPSCs but to a significantly lesser degree. The receptor subtypes mediating these effects were examined using specific 5-HT agonists and antagonists. A 5-HT1 agonist, 5-carboxamidotryptamine, produced EPSC depression but did not induce spontaneous IPSCs. A 5-HT2 agonist, alpha-Me-5-HT, reproduced all the observed effects of 5-HT (PSC depression as well as induction of spontaneous IPSCs), whereas a 5-HT2 antagonist, ketanserin, blocked the induction of spontaneous IPSCs. Therefore induction of spontaneous IPSCs is mediated by 5-HT2 receptors, whereas both 5-HT1 and 5-HT2 receptor types contribute to PSC depression. Serotonergic modulation of LSO synapses may have consequences for both developmental plasticity and auditory function. Serotonergic induction of IPSCs was observed primarily in young animals and thus may represent a mechanism for amplifying the activity of inhibitory synapses in LSO during a period of use-dependent plasticity in postnatal development. PSC depression, which preferentially affects excitation, is a potential mechanism for modulation of ILD tuning. PMID- 10368395 TI - Unmyelinated afferents constitute a second system coding tactile stimuli of the human hairy skin. AB - Impulses were recorded from unmyelinated afferents innervating the forearm skin of human subjects using the technique of microneurography. Units responding to innocuous skin deformation were selected. The sample (n = 38) was split into low threshold units (n = 27) and high-threshold units (n = 11) on the basis of three distinctive features, i.e., thresholds to skin deformation, size of response to innocuous skin deformation, and differential response to sharp and blunt stimuli. The low-threshold units provisionally were denoted tactile afferents on the basis of their response properties, which strongly suggest that they are coding some feature of tactile stimuli. They exhibited, in many respects, similar functional properties as described for low-threshold C-mechanoreceptive units in other mammals. However, a delayed acceleration, not previously demonstrated, was observed in response to long-lasting innocuous indentations. It was concluded that human hairy skin is innervated by a system of highly sensitive mechanoreceptive units with unmyelinated afferents akin to the system previously described in other mammals. The confirmation that the system is present in the forearm skin and not only in the face area where it first was identified suggests a largely general distribution although there are indications that the tactile C afferents may be lacking in the very distal parts of the limbs. The functional role of the system remains to be assessed although physiological properties of the sense organs invite to speculations that the slow tactile system might have closer relations to limbic functions than to cognitive and motor functions. PMID- 10368396 TI - Influence of gaze rotation on the visual response of primate MSTd neurons. AB - When we move forward, the visual image on our retina expands. Humans rely on the focus, or center, of this expansion to estimate their direction of heading and, as long as the eyes are still, the retinal focus corresponds to the heading. However, smooth rotation of the eyes adds nearly uniform visual motion to the expanding retinal image and causes a displacement of the retinal focus. In spite of this, humans accurately judge their heading during pursuit eye movements and during active, smooth head rotations even though the retinal focus no longer corresponds to the heading. Recent studies in macaque suggest that correction for pursuit may occur in the dorsal aspect of the medial superior temporal area (MSTd) because these neurons are tuned to the retinal position of the focus and they modify their tuning during pursuit to compensate partially for the focus shift. However, the question remains whether these neurons also shift focus tuning to compensate for smooth head rotations that commonly occur during gaze tracking. To investigate this question, we recorded from 80 MSTd neurons while monkeys tracked a visual target either by pursuing with their eyes or by vestibulo-ocular reflex cancellation (VORC; whole-body rotation with eyes fixed in head and head fixed on body). VORC is a passive, smooth head rotation condition that selectively activates the vestibular canals. We found that neurons shift their focus tuning in a similar way whether focus displacement is caused by pursuit or by VORC. Across the population, compensation averaged 88 and 77% during pursuit and VORC, respectively (tuning shift divided by the retinal focus to true heading difference). Moreover the degree of compensation during pursuit and VORC was correlated in individual cells (P < 0.001). Finally neurons that did not compensate appreciably tended to be gain-modulated during pursuit and VORC and may constitute an intermediate stage in the compensation process. These results indicate that many MSTd cells compensate for general gaze rotation, whether produced by eye-in-head or head-in-world rotation, and further implicate MSTd as a critical stage in the computation of heading. Interestingly vestibular cues present during VORC allow many cells to compensate even though humans do not accurately judge their heading in this condition. This suggests that MSTd may use vestibular information to create a compensated heading representation within at least a subpopulation of cells, which is accessed perceptually only when additional cues related to active head rotations are also present. PMID- 10368397 TI - Synaptic transmission in pair recordings from CA3 pyramidal cells in organotypic culture. AB - We performed simultaneous whole cell recordings from pairs of monosynaptically coupled hippocampal CA3 pyramidal neurons in organotypic slices. Stimulation of an action potential in a presynaptic cell resulted in an AMPA-receptor-mediated excitatory postsynaptic current (EPSC) in the postsynaptic cell that averaged approximately 34 pA. The average size of EPSCs varied in amplitude over a 20-fold range across different pairs. Both paired-pulse facilitation and depression were observed in the synaptic current in response to two presynaptic action potentials delivered 50 ms apart, but the average usually was dominated by depression. In addition, the amplitude of the second EPSC depended on the amplitude of the first EPSC, indicating competition between successive events for a common resource that is not restored within the 50-ms interpulse interval. Variation in the synaptic strength among pairs could arise from a variety of sources. Our data from anatomic reconstruction, 1/CV2 analysis, paired-pulse analysis, and manipulations of calcium/magnesium ratio suggest that differences in quantal size and release probability do not appear to vary sufficiently to fully account for the observed differences in amplitude. Thus it seems most likely that the variability in EPSC amplitude between pairs arises primarily from differences in the number of functional synapses. Injections of the calcium chelator bis-(o-aminophenoxy)-N, N,N',N'-tetraacetic acid into the presynaptic neuron resulted in a rapid and nearly complete block of transmission, whereas injection of the slower-acting chelator EGTA resulted in a variable and partial block. In addition to demonstrating the feasibility of manipulating the intracellular presynaptic environment by injection into the presynaptic soma, these data, and the EGTA results in particular may suggest variability in the linkage between calcium entry sites an release sites in these synapses. PMID- 10368398 TI - Characteristics of simian adaptation fields produced by behavioral changes in saccade size and direction. AB - The gain of saccadic eye movements can be altered gradually by moving targets either forward or backward during targeting saccades. If the gain of saccades to targets of only one size is adapted, the gain change generalizes or transfers only to saccades with similar vectors. In this study, we examined the spatial extent of such saccadic size adaptation, i.e., the gain adaptation field. We also attempted to adapt saccade direction by moving the target orthogonally during the targeting saccade to document the extent of a direction or cross-axis adaptation field. After adaptive gain decreases of horizontal saccades to 15 degrees target steps, >82% of the gain reduction transferred to saccades to 25 degrees horizontal target steps but only approximately 30% transferred to saccades to 5 degrees steps. For the horizontal component of oblique saccades to target steps with 15 degrees horizontal components and 10 degrees upward or downward vertical components, the transfer was similar at 51 and 60%, respectively. Thus the gain decrease adaptation field was quite asymmetric in the horizontal dimension but symmetric in the vertical dimension. Although gain increase adaptation produced a smaller gain change (13% increase for a 30% forward adapting target step) than did gain decrease adaptation (20% decrease for a 30% backward adapting target step), the spatial extent of gain transfer was quite similar. In particular, the gain increase adaptation field displayed asymmetry in the horizontal dimension (58% transfer to 25 degrees saccades but only 32% transfer to 5 degrees saccades) and symmetry in the vertical direction (50% transfer to the horizontal component of 10 degrees upward and 40% transfer to 10 degrees downward oblique saccades). When a 5 degrees vertical target movement was made to occur during a saccade to a horizontal 10 degrees target step, a vertical component gradually appeared in saccades to horizontal targets. More than 88% of the cross-axis change in the vertical component produced in 10 degrees saccades transferred to 20 degrees saccades but only 12% transferred to 4 degrees saccades. The transfer was similar to the vertical component of oblique saccades to target steps with either 10 degrees upward (46%) or 10 degrees downward (46%) vertical components. Therefore both gain and cross-axis adaptation fields have similar spatial profiles. These profiles resemble those of movement fields of neurons in the frontal eye fields and superior colliculus. How those structures might participate in the adaptation process is considered in the DISCUSSION. PMID- 10368399 TI - Blockade of GABAA receptors facilitates induction of NMDA receptor-independent long-term potentiation. AB - An N-methyl-D-aspartate (NMDA)-independent form of long-term potentiation (LTP), which depends on postsynaptic, voltage-dependent calcium channels (VDCCs), has been demonstrated in area CA1 of hippocampus. GABA acting at GABAA receptors limits postsynaptic depolarization during LTP induction. Blockade of GABAA receptors should therefore enhance activation of postsynaptic VDCCs and facilitate the induction of this NMDA receptor-independent, VDCC-dependent LTP. In agreement with this hypothesis, pharmacological blockade of GABAA receptors in the in vitro rat hippocampal slice increased the magnitude of LTP resulting from a normally effective, high-frequency (200 Hz) tetanic stimulation protocol. In addition, GABAA receptor blockade allowed a lower frequency (25 Hz) and normally ineffective tetanic stimulation protocol to induce this form of LTP. Intracellular recordings from CA1 pyramidal cells revealed that blocking GABAA receptors during tetanic stimulation allowed greater postsynaptic depolarization, increased the number of postsynaptic action potentials fired during the tetanization, and also increased the duration of synaptically evoked action potentials. To mimic the increased action potential firing observed when GABAA receptors were blocked, we paired 25-Hz antidromic stimulation with 25-Hz orthodromic stimulation. Paired antidromic + orthodromic 25-Hz stimulation induced NMDA receptor-independent LTP, whereas neither antidromic nor orthodromic stimulation alone induced LTP. Increased action potential firing can therefore at least partially account for the facilitation of NMDA receptor-independent LTP caused by blockade of GABAA receptors. This conclusion is consistent with prior studies demonstrating that action potentials are particularly effective stimuli for the gating of VDCCs in CA1 pyramidal cell dendrites. PMID- 10368400 TI - Testing the classification of static gamma axons using different patterns of random stimulation. AB - The possibility of using randomly generated stimulus intervals (with a Poisson distribution) to identify the type(s) of intrafusal fiber activated by the stimulation of single static gamma axons was tested in Peroneus tertius muscle spindles of anesthetized cats. Three patterns of random stimulation with different values of mean intervals [20 +/- 4. 47, 30 +/- 8.94, and 40 +/- 8.94 (SD) ms] were used. Single static gamma axons activating, in single spindles, either the bag2 fiber alone or the chain fibers alone or both types of intrafusal fiber were prepared. Responses of spindle primary endings elicited by the stimulation of gamma axons were recorded from Ia fibers in cut dorsal root filaments. Cross-correlograms between stimuli and spikes of the primary ending responses, autocorrelograms, interval histograms of responses, and stimulations were built. The characteristics of cross-correlograms were found to be related not only to the type of intrafusal muscle fibers activated but also to the parameters of the stimulation. Moreover some cross-correlograms with similar characteristics were produced by the activation of different intrafusal muscle fibers. It also was observed that, whatever the type of intrafusal muscle fiber activated, cross-correlograms could exhibit oscillations after an initial peak, provided the extent in frequency of the primary ending response was small; these oscillations arise in part from the autocorrelation of the primary ending responses. Therefore, cross-correlograms obtained during random stimulation of static gamma axons cannot be used for unequivocally identifying the type(s) of intrafusal muscle fiber these axons supply. PMID- 10368401 TI - Receptive fields and binaural interactions for virtual-space stimuli in the cat inferior colliculus. AB - Sound localization depends on multiple acoustic cues such as interaural differences in time (ITD) and level (ILD) and spectral features introduced by the pinnae. Although many neurons in the inferior colliculus (IC) are sensitive to the direction of sound sources in free field, the acoustic cues underlying this sensitivity are unknown. To approach this question, we recorded the responses of IC cells in anesthetized cats to virtual space (VS) stimuli synthesized by filtering noise through head-related transfer functions measured in one cat. These stimuli not only possess natural combinations of ITD, ILD, and spectral cues as in free field but also allow precise control over each cue. VS receptive fields were measured in the horizontal and median vertical planes. The vast majority of cells were sensitive to the azimuth of VS stimuli in the horizontal plane for low to moderate stimulus levels. Two-thirds showed a "contra preference" receptive field, with a vigorous response on the contralateral side of an edge azimuth. The other third of receptive fields were tuned around a best azimuth. Although edge azimuths of contra-preference cells had a broad distribution, best azimuths of tuned cells were near the midline. About half the cells tested were sensitive to the elevation of VS stimuli along the median sagittal plane by showing either a peak or a trough at a particular elevation. In general receptive fields for VS stimuli were similar to those found in free-field studies of IC neurons, suggesting that VS stimulation provided the essential cues for sound localization. Binaural interactions for VS stimuli were studied by comparing responses to binaural stimulation with responses to monaural stimulation of the contralateral ear. A majority of cells showed either purely inhibitory (BI) or mixed facilitatory/inhibitory (BF&I) interactions. Others showed purely facilitatory (BF) or no interactions (monaural). Binaural interactions were correlated with azimuth sensitivity: most contra-preference cells had either BI or BF&I interactions, whereas tuned cells were usually BF. These correlations demonstrate the importance of binaural interactions for azimuth sensitivity. Nevertheless most monaural cells were azimuth-sensitive, suggesting that monaural cues also play a role. These results suggest that the azimuth of a high-frequency sound source is coded primarily by edges in azimuth receptive fields of a population of ILD-sensitive cells. PMID- 10368403 TI - Physiological properties of neurons in the ventral nucleus of the lateral lemniscus of the rat: intrinsic membrane properties and synaptic responses. AB - The physiological properties including current-voltage relationships, firing patterns, and synaptic responses of the neurons in the ventral nucleus of the lateral lemniscus (VNLL) were studied in brain slices taken through the young rat's (17-37 days old) auditory brain stem. Intracellular recordings were made from VNLL neurons, and synaptic potentials were elicited by electrical stimulation of the lateral lemniscus ventral to the VNLL. Current-voltage relations and firing patterns were tested by recording the electrical potentials produced by intracellular injection of positive and negative currents. There were two types of VNLL neurons (type I and II) that exhibited different current voltage relationships. In response to negative current, both type I and II neurons produced a graded hyperpolarization. Type I neurons responded to positive current with a graded depolarization and multiple action potentials the number of which was related to the strength of the current injected. The current-voltage relations of type I neurons were nearly linear. Type II neurons responded to positive current with a limited depolarization and only one or a few action potentials. The current-voltage relations of type II neurons were nonlinear near the resting potential. The membrane properties of the type II VNLL neurons may play an important role for processing information about time of onset of a sound. Type I neurons showed three different firing patterns, i.e., regular, onset-pause and adaptation, in response to small positive current. The onset-pause and adaptation patterns could become sustained when a large current was injected. The regular, onset-pause, and adaptation patterns in type I neurons and the onset pattern in type II neurons resemble "chopper," "pauser, " "primary-like," and "on" responses, respectively, as defined in in vivo VNLL studies. The results suggest that different responses to acoustic stimulation could be attributed to intrinsic membrane properties of VNLL neurons. Many VNLL neurons responded to stimulation of the lateral lemniscus with excitatory or inhibitory responses or both. Excitatory and inhibitory responses showed interaction, and the output of the synaptic integration depended on the relative strength of excitatory and inhibitory responses. Neurons with an onset-pause firing pattern were more likely to receive mixed excitatory and inhibitory inputs from the lower auditory brain stem. PMID- 10368402 TI - Maturation of neuromuscular transmission during early development in zebrafish. AB - We have examined the rapid development of synaptic transmission at the neuromuscular junction (NMJ) in zebrafish embryos and larvae by patch-clamp recording of spontaneous miniature endplate currents (mEPCs) and single acetylcholine receptor (AChR) channels. Embryonic (24-36 h) mEPCs recorded in vivo were small in amplitude (<50 pA). The rate of mEPCs increased in larvae (3.5 fold increase measured by 6 days), and these mEPCs were mostly of larger amplitude (10-fold on average) with (10 times longer than that during visual fixation. When subjects performed horizontal smooth-pursuit eye movements that ended (i.e., 0 gaze velocity) just before the head rotation, the gaze velocity perturbation at the onset of head rotation was absent or small. The initial gain of the VOR had been significantly increased by the prior pursuit movements for all subjects (P < 0.05; mean increase of 11%). In four subjects, we determined that horizontal saccades and smooth tracking of a head-fixed target (VOR cancellation with eye stationary in the orbit) also increased the initial VOR gain (by a mean of 13%) during subsequent head rotations. However, after vertical saccades or smooth pursuit, the initial gaze perturbation caused by a horizontal head rotation was similar to that which occurred after fixation of a stationary target. We conclude that the initial gain of the VOR during a sudden horizontal head rotation is increased by prior horizontal, but not vertical, visually mediated gaze shifts. We postulate that this "priming" effect of a prior gaze shift on the gain of the VOR occurs at the level of the velocity inputs to the neural integrator subserving horizontal eye movements, where gaze-shifting commands and vestibular signals converge. PMID- 10368406 TI - Modulation of stretch reflexes during imposed walking movements of the human ankle. AB - Our overall objectives were to examine the role of peripheral afferents from the ankle in modulating stretch reflexes during imposed walking movements and to assess the mechanical consequences of this reflex activity. Specifically we sought to define the changes in the electromyographic (EMG) and mechanical responses to a stretch as a function of the phase of the step cycle. We recorded the ankle position of a normal subject walking on a treadmill at 3 km/h and used a hydraulic actuator to impose the same movements on supine subjects generating a constant level of ankle torque. Small pulse displacements, superimposed on the simulated walking movement, evoked stretch reflexes at different phases of the cycle. Three major findings resulted: 1) soleus reflex EMG responses were influenced strongly by imposed walking movements. The response amplitude was substantially smaller than that observed during steady-state conditions and was modulated throughout the step cycle. This modulation was qualitatively similar to that observed during active walking. Because central factors were held constant during the imposed walking experiments, we conclude that peripheral mechanisms were capable of both reducing the amplitude of the reflex EMG and producing its modulation throughout the movement. 2) Pulse disturbances applied from early to midstance of the imposed walking cycle generated large reflex torques, suggesting that the stretch reflex could help to resist unexpected perturbations during this phase of walking. In contrast, pulses applied during late stance and swing phase generated little reflex torque. 3) Reflex EMG and reflex torque were modulated differently throughout the imposed walking cycle. In fact, at the time when the reflex EMG response was largest, the corresponding reflex torque was negligible. Thus movement not only changes the reflex EMG but greatly modifies the mechanical output that results. PMID- 10368408 TI - Turning strategies during human walking. AB - The mechanisms involved in rapidly turning during human walking were studied. Subjects were asked to walk at a comfortable speed and to turn toward the instructed direction as soon as they felt an electrical stimulus to the superficial peroneal nerve. Stimuli were presented repeatedly at random over 10- to 15-min periods of walking for turning in both directions. Electromyograms (EMGs), joint angular movements of the right leg, and forces under both feet were recorded. The step cycle was divided into 16 parts, and the responses to stimuli in each part were analyzed separately. Two turning strategies were used, depending on which leg was placed in front for braking. For example, to turn to the right when the right foot was placed in front, subjects generally altered direction by spinning the body around the right foot (spin turn). To turn left when the right foot was in front, subjects shifted weight to the right leg, externally rotated the left hip, stepped onto the left leg, and continued turning until the right leg stepped in the new direction (step turn). The step turn is easy and stable because the base of support during the turn is much wider than in the spin turn, so some subjects used it in all parts of the cycle. Initially, the deceleration of walking is similar to a rapid stopping task, which has been previously examined. The deceleration mechanism involves a sequence of distal-to proximal activation of muscles on one side of the body (soleus, biceps femoris, and erector spinae). This pattern is similar to the "ankle strategy" used in postural control during forward sway. The control of foot placement in the swing leg and muscle activities for rotating the trunk in the stance leg occurred within a step after the cue. The action of ankle inverters and elevation of the pelvis by activity of gluteus medius may contribute to the control of trunk rotation. This activity was closely related to the timing of the opposite foot strike, independent of the part of the step cycle when the stimulus was applied. In most subjects, the turn was completed without resetting the underlying walking rhythm. This first EMG analysis of rapid turning shows how common strategies for postural sway and stopping can be combined with one of two turning strategies. This simplifies the complex task of turning at a random time in the step cycle. PMID- 10368407 TI - Group I mGluR activation causes voltage-dependent and -independent Ca2+ rises in hippocampal pyramidal cells. AB - Application of the metabotropic glutamate receptor (mGluR) agonist (1S, 3R)-1 aminocyclopentane-1,3-dicarboxylic acid (ACPD) or the selective group I mGluR agonist (S)-3,5-dihydroxyphenylglycine (DHPG) depolarized both CA3 and CA1 pyramidal cells in guinea pig hippocampal slices. Simultaneous recordings of voltage and intracellular Ca2+ levels revealed that the depolarization was accompanied by a biphasic elevation of intracellular Ca2+ concentration ([Ca2+]i): a transient calcium rise followed by a delayed, sustained elevation. The transient [Ca2+]i rise was independent of the membrane potential and was blocked when caffeine was added to the perfusing solution. The sustained [Ca2+]i rise appeared when membrane depolarization reached threshold for voltage-gated Ca2+ influx and was suppressed by membrane hyperpolarization. The depolarization was associated with an increased input resistance and persisted when either the transient or sustained [Ca2+]i responses was blocked. mGluR-mediated voltage and [Ca2+]i responses were blocked by (+)-alpha-methyl-4-carboxyphenylglycine (MCPG) or (S)-4-carboxy-3-hydroxyphenylglycine (4C3HPG). These data suggest that in both CA3 and CA1 hippocampal cells, activation of group I mGluRs produced a biphasic accumulation of [Ca2+]i via two paths: a transient release from intracellular stores, and subsequently, by influx through voltage-gated Ca2+ channels. The concurrent mGluR-induced membrane depolarization was not caused by the [Ca2+]i rise. PMID- 10368409 TI - Reciprocal synaptic interactions between rod bipolar cells and amacrine cells in the rat retina. AB - Reciprocal synaptic transmission between rod bipolar cells and presumed A17 amacrine cells was studied by whole cell voltage-clamp recording of rod bipolar cells in a rat retinal slice preparation. Depolarization of a rod bipolar cell evoked two identifiable types of Ca2+ current, a T-type current that activated at about -70 mV and a current with L-type pharmacology that activated at about -50 mV. Depolarization to greater than or equal to -50 mV also evoked an increase in the frequency of postsynaptic currents (PSCs). The PSCs reversed at approximately ECl (the chloride equilibrium potential), followed changes in ECl, and were blocked by gamma-aminobutyric acidA (GABAA) and GABAC receptor antagonists and thus were identified as GABAergic inhibitory PSCs (IPSCs). Bipolar cells with cut axons displayed the T-type current but lacked an L-type current and depolarization-evoked IPSCs. Thus L-type Ca2+ channels are placed strategically at the axon terminals to mediate transmitter release from rod bipolar cells. The IPSCs were blocked by the non-N-methyl-D-aspartate (non-NMDA) receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione, indicating that non-NMDA receptors mediate the feed-forward bipolar-to-amacrine excitation. The NMDA receptor antagonist 3 ((RS)-2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid had no consistent effect on the depolarization-evoked IPSCs, indicating that activation of NMDA receptors is not essential for the feedforward excitation. Tetrodotoxin (a blocker of voltage-gated Na+ channels) reversibly suppressed the reciprocal response in some cells but not in others, indicating that graded potentials are sufficient for transmitter release from A17 amacrine cells, but suggesting that voltage-gated Na+ channels, under some conditions, can contribute to transmitter release. PMID- 10368410 TI - Voltage-activated potassium outward currents in two types of spider mechanoreceptor neurons. AB - We studied the properties of voltage-activated outward currents in two types of spider cuticular mechanoreceptor neurons to learn if these currents contribute to the differences in their adaptation properties. Both types of neurons adapt rapidly to sustained stimuli, but type A neurons usually only fire one or two action potentials, whereas type B neurons can fire bursts lasting several hundred milliseconds. We found that both neurons had two outward current components, 1) a transient current that activated rapidly when stimulated from resting potential and inactivated with maintained stimuli and 2) a noninactivating outward current. The transient outward current could be blocked by 5 mM tetraethylammonium chloride, 5 mM 4-aminopyridine, or 100 microM quinidine, but these blockers also reduced the amplitude of the noninactivating outward current. Charybdotoxin or apamin did not have any effect on the outward currents, indicating that Ca2+ activated K+ currents were not present or not inhibited by these toxins. The only significant differences between type A and type B neurons were found in the half maximal activation (V50) values of both currents. The transient current had a V50 value of 9. 6 mV in type A neurons and -13.1 mV in type B neurons, whereas the V50 values of noninactivating outward currents were -48.9 mV for type A neurons and -56.7 mV for type B neurons. We conclude that, although differences in the activation kinetics of the voltage-activated K+ currents could contribute to the difference in the adaptation behavior of type A and type B neurons, they are not major factors. PMID- 10368411 TI - Response attenuation during coincident afferent excitatory inputs. AB - The linearity of the synaptic summation of two unitary excitatory synaptic events was investigated during whole cell recordings from retinal target neurons in an eye-attached isolated brain stem preparation. Pairs of unitary excitatory postsynaptic potentials (EPSPs) were evoked by bipolar stimulation electrodes that were directed to two distinct foci on the retinal surface based on the visual receptive field boundaries. The interval between stimulation of each retinal site was incremented by 0.5-1 ms to quantify the time course of nonlinear summation using an exponential fit. Response facilitation was never observed; however, the coincident arrival of synaptic inputs caused a response attenuation in 26 of the 37 pairs studied. Twelve of the 26 pairs had time constants of their attenuation that were similar to the time constants of the decaying phases of the first EPSPs of each pair. This suggests that the attenuation of these 12 pairs may be entirely due to voltage-dependent mechanisms, such as a reduction in driving force or a change of the activity of voltage-sensitive channels. On the other hand, the 14 other pairs had their time constant of attenuation shorter than the time constants of the decaying phase of the first EPSP. In fact, the attenuation time constants were often closer to the time constants of the decaying phases of the first excitatory postsynaptic currents of each pair. This finding suggests that the attenuation of these 14 pairs involve a shunting mechanism due to the opening of synaptic channels. The presence of this conductance-dependent mechanism is supported by the finding of asymmetric effects on the time course of attenuation when the stimulation sequence was reversed. These results are discussed in terms of the processing by neurons of coincident excitatory inputs onto spatially distinct points of their dendritic trees. PMID- 10368412 TI - Differentiating cortical areas related to pain perception from stimulus identification: temporal analysis of fMRI activity. AB - In a recent functional magnetic resonance imaging study (fMRI), we reported the cortical areas activated in a thermal painful task and compared the extent of overlap between this cortical network and those activated during a vibrotactile task and a motor task. In the present study we examine the temporal properties of the cortical activations for all three tasks and use linear systems identification techniques to functionally differentiate the cortical regions identified in the painful thermal task. Cortical activity was examined in the contralateral middle third of the brain of 10 right-handed subjects, using echo planar imaging and a surface coil. In another eight subjects the temporal properties of the thermal task were examined psychophysically. The fMRI impulse response function was estimated from the cortical activations in the vibrotactile and motor tasks and shown to correspond to earlier reports. Given the fMRI impulse response function and the time courses for the thermal stimulus and the associated pain ratings, predictor functions were generated. The correlation between these predictor functions and cortical activations in the painful thermal task indicated a gradual transition of information processing anteroposteriorly in the parietal cortex. Within this region, activity in the anterior areas more closely reflected thermal stimulus parameters, whereas activity more posteriorly was better related to the temporal properties of pain perception. Insular cortex at the level of the anterior commissure was the region best related to the thermal stimulus, and Brodmann's area 5/7 was the region best related to the pain perception. The functional implications of these observations are discussed. PMID- 10368413 TI - In situ and in vitro identification and characterization of cardiac ganglion neurons in the crab, Carcinus maenas. AB - The aim of this study was to investigate the intrinsic membrane properties and hormonal responses of individual central pattern generating neurons in the cardiac ganglion of the shore crab Carcinus maenas. Because the cardiac ganglion in this crustacean species is buried within the heart musculature and is therefore inaccessible for direct morphological and electrophysiological analysis, we developed two novel in vitro preparations. First, to make the ganglion accessible, we established a brief enzymatic treatment procedure that enabled us to isolate the entire cardiac ganglion, in the absence of muscle tissue. Second, a cell culture procedure was developed to isolate individual neurons in vitro. With the use of both isolated ganglionic and neuronal cell culture techniques, this study provides the first direct account of the neuroanatomy of the cardiac ganglion in shore crabs. We demonstrate that cultured neurons not only survived the isolation procedures, but that they also maintained their intrinsic membrane and transmitter response properties, similar to those seen in the intact ganglion. Specifically, we tested the peptides proctolin, crustacean cardioactive peptide, the FLRFamide-related peptide F2, and an amine (serotonin) on both isolated ganglion and in vitro culture neurons. We measured changes in neuronal burst rate, burst amplitude, pacemaker slope, and membrane potential oscillation amplitude in response to the above four hormones. Each hormone either increased neuronal activity in spontaneously bursting neurons, or induced a bursting pattern in quiescent cells. The in vitro cell culture system developed here now provides us with an excellent opportunity to elucidate cellular, synaptic and hormonal mechanisms by which cardiac activity is generated in shore crabs. PMID- 10368414 TI - Reciprocal interactions in the turtle hindlimb enlargement contribute to scratch rhythmogenesis. AB - We examined interactions between the spinal networks that generate right and left rostral scratch motor patterns in turtle hindlimb motoneurons before and after transecting the spinal cord within the anterior hindlimb enlargement. Our results provide evidence that reciprocal inhibition between hip circuit modules can generate hip rhythmicity during the rostral scratch reflex. "Module" refers here to the group of coactive motoneurons and interneurons that controls either flexion or extension of the hip on one side and coordinates that activity with synergist and antagonist motor pools in the same limb and in the contralateral limb. The "bilateral shared core" hypothesis states that hip flexor and extensor (HF and HE) circuit modules interact via crossed and uncrossed spinal pathways: HF modules make reciprocal inhibitory connections with contralateral HF and ipsilateral HE modules and mutual excitatory connections with contralateral HE modules. It is currently unclear how much reciprocal inhibition between modules contributes to scratch rhythmogenesis. To address this issue, fictive scratch motor patterns were recorded bilaterally as electroneurograms from HF, HE, knee extensor (KE), and respiratory (d.D8) muscle nerves in immobilized animals. D3 end (low-spinal) preparations had intact spinal cords posterior to a complete D2 D3 transection. Unilateral stimulation of rostral scratch in D3-end turtles elicited rhythmic alternation between ipsilateral HF and HE bursts in most cycles; consecutive HF bursts were separated by complete silent (HF-OFF ) periods. D3-D9 and D3-D8 preparations received a second spinal transection at the caudal end of segment D9 or D8, respectively, within the anterior hindlimb enlargement. This second transection disconnected most HE circuitry (located mainly in segments D10-S2 of the posterior enlargement) from the rostral scratch network and thereby reduced the HE-associated inhibition of HF circuitry. Unilateral stimulation of rostral scratch in most D3-D9 and D3-D8 preparations evoked rhythmic or weakly modulated ipsilateral HF discharge without HF-OFF periods between bursts and without ipsilateral HE activity in the majority of cycles. In contrast, bilateral stimulation in D3-D9 and D3-D8 preparations reconstructed the HF-OFF periods, increased HF rhythmicity (assessed by fast Fourier transform power spectra and autocorrelation analyses), and reestablished weak HE-phase motoneuron activity. We suggest that bilateral stimulation produced these effects by simultaneously activating reciprocally inhibitory hip modules on opposite sides (right and left HF) and the same side (HF and residual ipsilateral HE circuitry). Our data support the hypothesis that reciprocal inhibition can contribute to spinal rhythmogenesis during the scratch reflex. PMID- 10368415 TI - SI neuron response variability is stimulus tuned and NMDA receptor dependent. AB - Skin brushing stimuli were used to evoke spike discharge activity in single skin mechanoreceptive afferents (sMRAs) and anterior parietal cortical (SI) neurons of anesthetized monkeys (Macaca fascicularis). In the initial experiments 10-50 presentations of each of 8 different stimulus velocities were delivered to the linear skin path from which maximal spike discharge activity could be evoked. Mean rate of spike firing evoked by each velocity (MFR) was computed for the time period during which spike discharge activity exceeded background, and an across presentations estimate of mean firing rate (MFR) was generated for each velocity. The magnitude of the trial-by-trial variation in the response (estimated as CV; where CV = standard deviation in MFR/MFR) was determined for each unit at each velocity. MFR for both sMRAs and SI neurons (MFRsMRA and MFRSI, respectively) increased monotonically with velocity over the range 1-100 cm/s. At all velocities the average estimate of intertrial response variation for SI neurons (CVSI) was substantially larger than the corresponding average for sMRAs (CVsMRA). Whereas CVsMRA increased monotonically over the range 1-100 cm/s, CVSI decreased progressively with velocity over the range 1-10 cm/s, and then increased with velocity over the range 10-100 cm/s. The position of the skin brushing stimulus in the receptive field (RF) was varied in the second series of experiments. It was found that the magnitude of CVSI varied systematically with stimulus position in the RF: that is, CVSI was lowest for a particular velocity and direction of stimulus motion when the skin brushing stimulus traversed the RF center, and CVSI increased progressively as the distance between the stimulus path and the RF center increased. In the third series of experiments, either phencylidine (PCP; 100-500 microg/kg) or ketamine (KET; 0.5-7.5 mg/kg) was administered intravenously (iv) to assess the effect of block of N-methyl-D aspartate (NMDA) receptors on SI neuron intertrial response variation. The effects of PCP on both CVSI and MFRSI were transient, typically with full recovery occurring in 1-2 h after drug injection. The effects of KET on CVSI and MFRSI were similar to those of PCP, but were shorter in duration (15-30 min). PCP and KET administration consistently was accompanied by a reduction of CVSI. The magnitude of the reduction of CVSI by PCP or KET was associated with the magnitude of CVSI before drug administration: that is, the larger the predrug CVSI, the larger the reduction in CVSI caused by PCP or KET. PCP and KET exerted variable effects on SI neuron mean firing rate that could differ greatly from one neuron to the next. The results are interpreted to indicate that SI neuron intertrial response variation is 1) stimulus tuned (intertrial response variation is lowest when the skin stimulus moves at 10 cm/s and traverses the neuron's RF center) and 2) NMDA receptor dependent (intertrial response variation is least when NMDA receptor activity contributes minimally to the response, and increases as the contribution of NMDA receptors to the response increases). PMID- 10368416 TI - Long-range inhibition within the zebra finch song nucleus RA can coordinate the firing of multiple projection neurons. AB - The zebra finch forebrain song control nucleus RA (robust nucleus of the archistriatum) generates a phasic and temporally precise neural signal that drives vocal and respiratory motoneurons during singing. RA's output during singing predicts individual notes, even though afferent drive to RA from the song nucleus HVc is more tonic, and predicts song syllables, independent of the particular notes that comprise the syllable. Therefore RA's intrinsic circuitry transforms neural activity from HVc into a highly precise premotor output. To understand how RA's intrinsic circuitry effects this transformation, we characterized RA interneurons and projection neurons using intracellular recordings in brain slices. RA interneurons fired fast action potentials with steep current-frequency relationships and had small somata with thin aspinous processes that extended throughout large portions of the nucleus; the similarity of their fine processes to those labeled with a glutamic acid decarboxylase (GAD) antibody strongly suggests that these interneurons are GABAergic. Electrical stimulation revealed that RA interneurons receive excitatory inputs from RA's afferents, the lateral magnocellular nucleus of the anterior neostriatum (LMAN) and HVc, and from local axon collaterals of RA projection neurons. To map the functional connections that RA interneurons make onto RA projection neurons, we focally uncaged glutamate, revealing long-range inhibitory connections in RA. Thus these interneurons provide fast feed-forward and feedback inhibition to RA projection neurons and could help create the phasic pattern of bursts and pauses that characterizes RA output during singing. Furthermore, selectively activating the inhibitory network phase locks the firing of otherwise unconnected pairs of projection neurons, suggesting that local inhibition could coordinate RA output during singing. PMID- 10368417 TI - Stochastic nature of precisely timed spike patterns in visual system neuronal responses. AB - It is not clear how information related to cognitive or psychological processes is carried by or represented in the responses of single neurons. One provocative proposal is that precisely timed spike patterns play a role in carrying such information. This would require that these spike patterns have the potential for carrying information that would not be available from other measures such as spike count or latency. We examined exactly timed (1-ms precision) triplets and quadruplets of spikes in the stimulus-elicited responses of lateral geniculate nucleus (LGN) and primary visual cortex (V1) neurons of the awake fixating rhesus monkey. Large numbers of these precisely timed spike patterns were found. Information theoretical analysis showed that the precisely timed spike patterns carried only information already available from spike count, suggesting that the number of precisely timed spike patterns was related to firing rate. We therefore examined statistical models relating precisely timed spike patterns to response strength. Previous statistical models use observed properties of neuronal responses such as the peristimulus time histogram, interspike interval, and/or spike count distributions to constrain the parameters of the model. We examined a new stochastic model, which unlike previous models included all three of these constraints and unlike previous models predicted the numbers and types of observed precisely timed spike patterns. This shows that the precise temporal structures of stimulus-elicited responses in LGN and V1 can occur by chance. We show that any deviation of the spike count distribution, no matter how small, from a Poisson distribution necessarily changes the number of precisely timed spike patterns expected in neural responses. Overall the results indicate that the fine temporal structure of responses can only be interpreted once all the coarse temporal statistics of neural responses have been taken into account. PMID- 10368418 TI - Differential roles of NMDA and non-NMDA receptors in synaptic responses of neurons in nucleus tractus solitarii of the rat. AB - The relative role of N-methyl-D-aspartate (NMDA) and non-NMDA receptors in synaptic responses of neurons in caudal nucleus tractus solitarii (cNTS) was delineated by immunohistochemical and electrophysiologic experiments in rats. Double immunohistochemical staining in in vivo experiments revealed that approximately 80% of cNTS neurons that showed Fos-like immunoreactivity induced by baroreceptor activation were generally also immunoreactive to non-NMDA receptor subunits GluR1 or GluR2. On the other hand, only 20% of Fos-labeled cNTS neurons showed immunoreactivity to NMDA receptor subunits NMDAR1 or NMDAR2. Stimulation of the ipsilateral solitary tract at suprathreshold intensity in slice preparations induced Fos expression in the cNTS and evoked either a single action potential or a complex synaptic response consisting of an initial action potential followed by a secondary slow depolarization. In a majority (70%) of cNTS neurons that exhibited the complex synaptic response, both the initial and secondary components were eliminated reversibly by 6-cyano-7-nitroquinoxaline-2,3 dione (20 microM). This non-NMDA antagonist also inhibited the single action potential manifested by the other population of cNTS neurons. On the other hand, only the secondary slow depolarization was blocked by D(-)-2-amino-5 phosphonopentanoic acid (250 microM) or potentiated by NMDA (1.7 microM). Our results suggested that NMDA and non-NMDA receptors are involved differentially in the synaptic responses of cNTS neurons. Non-NMDA receptors may be distributed predominantly on a majority of the second-order cNTS neurons that may receive primary baroreceptor afferent inputs. On the other hand, NMDA receptors are located primarily on higher-order neurons, which may be connected reciprocally with the second-order cNTS neurons. PMID- 10368419 TI - Partial uncoupling of neurotransmitter release from [Ca2+]i by membrane hyperpolarization. AB - The dependence of evoked and asynchronous release on intracellular calcium ([Ca2+]i) and presynaptic membrane potential was examined in single-release boutons of the crayfish opener neuromuscular junction. When a single bouton was depolarized by a train of pulses, [Ca2+]i increased to different levels according to the frequency of stimulation. Concomitant measurements of evoked release and asynchronous release, from the same bouton, showed that both increased in a sigmoidal manner as a function of [Ca2+]i. When each of the depolarizing pulses was immediately followed by a hyperpolarizing pulse, [Ca2+]i was elevated to a lesser degree than in the control experiments, and the rate of asynchronous release and the quantal content were reduced; most importantly, evoked quantal release terminated sooner. The diminution of neurotransmitter release by the hyperpolarizing postpulse (HPP) could not be entirely accounted for by the reduction in [Ca2+]i. The experimental results are consistent with the hypothesis that the HPP reduces the sensitivity of the release machinery to [Ca2+]i, thereby not only reducing the quantal content but also terminating the quantal release process sooner. PMID- 10368420 TI - Seizure-induced cell death produced by repeated tetanic stimulation in vitro: possible role of endoplasmic reticulum calcium stores. AB - Seizures may cause brain damage due to mechanisms initiated by excessive excitatory synaptic transmission. One such mechanism is the activation of death promoting intracellular cascades by the influx and the perturbed homeostasis of Ca2+. The neuroprotective effects of preventing the entry of Ca2+ from voltage dependent Ca2+ channels, NMDA receptors, and non-NMDA receptors, is well known. Less clear is the contribution to excitotoxicity of Ca2+ released from endoplasmic reticulum (ER) stores. We produced epileptiform discharges in combined entorhinal cortex/hippocampus slices using repeated tetanic stimulation of the Schaffer collaterals and assessed cell death after 1, 3, or 12-14 h with gel electrophoresis of genomic DNA and immunohistologically using terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine 5'-triphosphate (dUTP) nick end labeling (TUNEL) staining. We manipulated ER Ca2+ stores using two conventional drugs, dantrolene, which blocks the Ca2+ release channel, and thapsigargin, which blocks sarco-endoplasmic reticulum Ca2+-ATPases resulting in depletion of ER Ca2+ stores. To monitor epileptogenesis, and to assess effects attributable to dantrolene and thapsigargin on normal synaptic transmission, extracellular potentials were recorded in stratum pyramidale of the CA1 region. Repeated tetanic stimulation reliably produced primary afterdischarge and spontaneous epileptiform discharges, which persisted for 14 h, the longest time recorded. We did not observe indications of cell death attributable to seizures with either method when assessed after 1 or 3 h; however, qualitatively more degraded DNA always was observed in tetanized slices from the 12- to 14-h group compared with time-matched controls. Consistent with these data was a significant, fourfold, increase in the percentage of TUNEL-positive cells in CA3, CA1, and entorhinal cortex in tetanized slices from the 12- to 14-h group (16. 5 +/- 4.4, 33.7 +/- 7.1, 11.6 +/- 2.1, respectively; means +/- SE; n = 7) compared with the appropriate time-matched control (4.1 +/- 2.2, 7.3 +/- 2.0, 2.8 +/- 0.9, respectively; n = 6). Dantrolene (30 microM; n = 5) and thapsigargin (1 microM; n = 4) did not affect significantly normal synaptic transmission, assessed by the amplitude of the population spike after 30 min of exposure. Dantrolene and thapsigargin also were without effect on the induction or the persistence of epileptiform discharges, but both drugs prevented seizure-induced cell death when assessed with gel electrophoresis. We suggest that Ca2+ entering a cell from the outside, in addition to the Ca2+ contributed from ryanodine-sensitive stores (i.e., Ca2+-induced Ca2+ release), may be necessary for seizure-induced cell death. PMID- 10368421 TI - Mesial motor areas in self-initiated versus externally triggered movements examined with fMRI: effect of movement type and rate. AB - The human frontomesial cortex reportedly contains at least four cortical areas that are involved in motor control: the anterior supplementary motor area (pre SMA), the posterior SMA (SMA proper, or SMA), and, in the anterior cingulate cortex, the rostral cingulate zone (RCZ) and the caudal cingulate zone (CCZ). We used functional magnetic resonance imaging (fMRI) to examine the role of each of these mesial motor areas in self-initiated and visually triggered movements. Healthy subjects performed self-initiated movements of the right fingers (self initiated task, SI). Each movement elicited a visual signal that was recorded. The recorded sequence of visual signals was played back, and the subjects moved the right fingers in response to each signal (visually triggered task, VT). There were two types of movements: repetitive (FIXED) or sequential (SEQUENCE), performed at two different rates: SLOW or FAST. The four regions of interest (pre SMA, SMA, RCZ, CCZ) were traced on a high-resolution MRI of each subject's brain. Descriptive analysis, consisting of individual assessment of significant activation, revealed a bilateral activation in the four mesial structures for all movement conditions, but SI movements were more efficient than VT movements. The more complex and more rapid the movements, the smaller the difference in activation efficiency between the SI and the VT tasks, which indicated an additional processing role of the mesial motor areas involving both the type and rate of movements. Quantitative analysis was performed on the spatial extent of the area activated and the percentage of change in signal amplitude. In the pre SMA, activation was more extensive for SI than for VT movements, and for fast than for slow movements; the extent of activation was larger in the ipsilateral pre-SMA. In the SMA, the difference was not significant in the extent and magnitude of activation between SI and VT movements, but activation was more extensive for sequential than for fixed movements. In the RCZ and CCZ, both the extent and magnitude of activation were larger for SI than for VT movements. In the CCZ, both indices of activation were also larger for sequential than for fixed movements, and for fast than for slow movements. These data suggest functional specificities of the frontomesial motor areas with respect not only to the mode of movement initiation (self-initiated or externally triggered) but also to the movement type and rate. PMID- 10368422 TI - Cellular properties of lateral spinal nucleus neurons in the rat L6-S1 spinal cord. AB - Conventional intracellular recordings were made from 26 lateral spinal nucleus (LSN) neurons in slices of L6-S1 spinal cord from 10- to 15-day-old rats. At rest, LSN neurons did not fire spontaneous action potentials. With injection of a positive current pulse, action potentials had an amplitude of 72 +/- 7 (SD) mV and duration at half-peak height of 0.75 +/- 0.22 ms. Action potentials were followed by an afterpotential. Most LSN neurons (13/17) exhibited only an afterhyperpolarization (AHP); four neurons exhibited both a fast and a slow AHP separated by an afterdepolarization (ADP). For LSN neurons that exhibited only an AHP, a slow ADP could be identified during bath application of apamin (100 nM). Four of 11 LSN neurons showed a postinhibitory rebound (PIR). Two types of PIR were noted, one with high threshold and low amplitude and the other with low threshold and high amplitude. The PIR with high amplitude was partially blocked in 0 mM Ca2+/high Mg2+ (10 mM) recording solution. Repetitive firing properties were examined in 17 LSN neurons. On the basis of the ratio of the slopes between initial instantaneous firing and steady-state firing frequencies, neurons with low spike frequency adaptation (SFA, 8/17) and high SFA (4/17) were identified. In addition, 2/17 LSN neurons exhibited biphasic repetitive firing patterns, which were composed of a fast SFA, delayed excitation, and low SFA; another two neurons showed only delayed excitation. Plateau potentials also were found in two LSN neurons. Dorsal root stimulation revealed that most LSN neurons (12/13) had polysynaptic postsynaptic potentials (PSP); only one neuron exhibited a monosynaptic PSP. Electrical stimulation of the dorsal root evoked prolonged discharges in low SFA neurons and a short discharge in high SFA neurons. Intrinsic properties were modulated by bath application of substance P (SP). Membrane potentials were depolarized in all eight LSN neurons tested, and membrane resistance was either increased (n = 3) or decreased (n = 2). Both instantaneous firing and steady-state firing were facilitated by SP. In addition, oscillation of membrane potentials were induced in three LSN neurons. These results demonstrate that LSN neurons exhibit a variety of intrinsic properties, which may significantly contribute to sensory processing, including nociceptive processing. PMID- 10368423 TI - Sweet taste responses of mouse chorda tympani neurons: existence of gurmarin sensitive and -insensitive receptor components. AB - Inhibitory effects of gurmarin (gur) on responses to sucrose and other sweeteners of single fibers of the chorda tympani nerve in C57BL mice were examined. Of 30 single fibers that strongly responded to 0. 5 M sucrose but were not or to lesser extent responsive to 0.1 M NaCl, 0.01 M HCl, and 0.02 M quinine HCl (sucrose-best fibers), 16 fibers showed large suppression of responses to sucrose and other sweeteners by lingual treatment with 4.8 microM (approximately 20 microg/ml) gur (suppressed to 4-52% of control: gur-sensitive fibers), whereas the remaining 14 fibers showed no such gur inhibition (77-106% of control: gur-insensitive fibers). In gur-sensitive fibers, responses to sucrose inhibited by gur recovered to approximately 70% of control responses after rinsing the tongue with 15 mM beta-cyclodextrin and were almost abolished by further treatment with 2% pronase. In gur-insensitive fibers, sucrose responses were not inhibited by gur, but were largely suppressed by pronase. These results suggest existence of two different receptor components for sweeteners with different susceptibilities to gur in mouse taste cells, one gur sensitive and the other gur insensitive. Taste cells possessing each component may be specifically innervated by a particular type of chorda tympani neurons. PMID- 10368424 TI - Sodium action potentials are not required for light-evoked release of GABA or glycine from retinal amacrine cells. AB - Although most CNS neurons require sodium action potentials (Na-APs) for normal stimulus-evoked release of classical neurotransmitters, many types of retinal and other sensory neurons instead use only graded potentials for neurotransmitter release. The physiological properties and information processing capacity of Na AP-producing neurons appear significantly different from those of graded potential neurons. To classify amacrine cells in this dichotomy, we investigated whether Na-APs, which are often observed in these cells, are required for functional light-evoked release of inhibitory neurotransmitters from these cells. We recorded light-evoked inhibitory postsynaptic currents (IPSCs) from retinal ganglion cells, neurons directly postsynaptic to amacrine cells, and applied TTX to block Na-APs. In control solution, TTX application always led to partial suppression of the light-evoked IPSC. To isolate release from glycinergic amacrine cells, we used either bicuculline, a GABAA receptor antagonist, or picrotoxin, a GABAA and GABAC receptor antagonist. TTX application only partially suppressed the glycinergic IPSC. To isolate release from GABAergic amacrine cells, we used the glycine receptor blocker strychnine. TTX application only partially suppressed the light-evoked GABAergic IPSC. Glycinergic and GABAergic amacrine cells did not obviously differ in the usage of Na-APs for release. These observations, in conjunction with previous studies of other retinal neurons, indicate that amacrine cells, taken as a class, are the only type of retinal neuron that uses both Na-AP-dependent and -independent modes for light-evoked release of neurotransmitters. These results also provide evidence for another parallel between the properties of retinal amacrine cells and olfactory bulb granule cells. PMID- 10368425 TI - Lack of AMPA receptor desensitization during basal synaptic transmission in the hippocampal slice. AB - Excitatory postsynaptic currents in the CA1 region of rat hippocampal slices are mediated primarily by alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors in response to synaptically released glutamate. Outside-out patches from pyramidal cells in this region have shown that AMPA receptors are desensitized by short (1 ms) pulses of glutamate. We have taken a number of approaches to ask whether synaptic receptors desensitize in response to synaptically released glutamate in the slice. Recordings with paired pulses and minimal stimulation conditions that are presumably activating only a single release site do not show evidence for desensitization. Furthermore, cyclothiazide, a drug that blocks desensitization, does not alter paired-pulse ratios even under conditions of high probability of release, which should maximize desensitization. These results suggest that synaptic receptors do not desensitize in response to synaptically released glutamate during basal synaptic transmission. PMID- 10368426 TI - Parallel activation of primary and secondary somatosensory cortices in human pain processing. AB - Cerebral processing of pain has been shown to involve primary (SI) and secondary (SII) somatosensory cortices. However, the temporal activation pattern of these cortices in nociceptive processing has not been demonstrated so far. We therefore used whole-head magnetoencephalography to record cortical responses to cutaneous laser stimuli in six healthy human subjects. By using selective nociceptive stimuli our results confirm involvement of contralateral SI and bilateral SII in human pain processing. Beyond they show for the first time simultaneous activation onset of contralateral SI and SII after approximately 130 ms, indicating parallel thalamocortical distribution of nociceptive information. This contrasts to the serial cortical organization of tactile processing in higher primates and instead corresponds to the parallel cortical organization in lower primates and nonprimates. Thus our finding suggests preservation of the basic mammalian parallel organizational scheme in human pain processing, whereas in the tactile modality parallel organization appears to be abandoned in favor of a serial processing scheme. Functionally, preservation of direct access to SII underscores the relevance of this area in human pain processing, probably reflecting an important role of SII in nociceptive learning and memory. PMID- 10368427 TI - Contribution of the cerebellar flocculus to gaze control during active head movements. AB - The flocculus and ventral paraflocculus are adjacent regions of the cerebellar cortex that are essential for controlling smooth pursuit eye movements and for altering the performance of the vestibulo-ocular reflex (VOR). The question addressed in this study is whether these regions of the cerebellum are more globally involved in controlling gaze, regardless of whether eye or active head movements are used to pursue moving visual targets. Single-unit recordings were obtained from Purkinje (Pk) cells in the floccular region of squirrel monkeys that were trained to fixate and pursue small visual targets. Cell firing rate was recorded during smooth pursuit eye movements, cancellation of the VOR, combined eye-head pursuit, and spontaneous gaze shifts in the absence of targets. Pk cells were found to be much less sensitive to gaze velocity during combined eye-head pursuit than during ocular pursuit. They were not sensitive to gaze or head velocity during gaze saccades. Temporary inactivation of the floccular region by muscimol injection compromised ocular pursuit but had little effect on the ability of monkeys to pursue visual targets with head movements or to cancel the VOR during active head movements. Thus the signals produced by Pk cells in the floccular region are necessary for controlling smooth pursuit eye movements but not for coordinating gaze during active head movements. The results imply that individual functional modules in the cerebellar cortex are less involved in the global organization and coordination of movements than with parametric control of movements produced by a specific part of the body. PMID- 10368428 TI - Physicochemical optimization in the genetic code origin as the number of codified amino acids increases. AB - We have assumed that the coevolution theory of genetic code origin (Wong JT, Proc Natl Acad Sci USA 72:1909-1912, 1975) is essentially correct. This theory makes it possible to identify at least 10 evolutionary stages through which genetic code organization might have passed prior to reaching its current form. The calculation of the minimization level of all these evolutionary stages leads to the following conclusions. (1) The minimization percentages increased linearly with the number of amino acids codified in the codes of the various evolutionary stages when only the sense changes are considered in the analysis. This seems to favor the physicochemical theory of genetic code origin even if, as discussed in the paper, this observation is also compatible with the coevolution theory. (2) For the first seven evolutionary stages of the genetic code, this trend is less clear and indeed is inverted when we consider the global optimisation of the codes due to both sense changes and synonymous changes. This inverse correlation between minimization percentages and the number of amino acids codified in the codes of the intermediate stages seems to favor neither the physicochemical nor the stereochemical theories of genetic code origin, as it is in the early and intermediate stages of code development that these theories would expect minimization to have played a crucial role, and this does not seem to be the case. However, these results are in agreement with the coevolution theory, which attributes a role to the physicochemical properties of amino acids that, while important, is nevertheless subordinate to the mechanism which concedes codons from the precursor amino acids to the product amino acids as the primary factor determining the evolutionary structuring of the genetic code. The results are therefore discussed in the context of the various theories proposed to explain genetic code origin. PMID- 10368429 TI - A probabilistic treatment of phylogeny and sequence alignment. AB - Carrying out simultaneous tree-building and alignment of sequence data is a difficult computational task, and the methods currently available are either limited to a few sequences or restricted to highly simplified models of alignment and phylogeny. A method is given here for overcoming these limitations by Bayesian sampling of trees and alignments simultaneously. The method uses a standard substitution matrix model for residues together with a hidden Markov model structure that allows affine gap penalties. It escapes the heavy computational burdens of other models by using an approximation called the "*" rule, which replaces missing data by a sum over all possible values of variables. The behavior of the model is demonstrated on test sets of globins. PMID- 10368430 TI - Enhanced evolvability in immunoglobulin V genes under somatic hypermutation. AB - Darwinian theory requires that mutations be produced in a nonanticipatory manner; it is nonetheless consistent to suggest that mutations that have repeatedly led to nonviable phenotypes would be introduced less frequently than others-if under appropriate genetic control. Immunoglobulins produced during infection acquire point mutations that are subsequently selected for improved binding to the eliciting antigen. We and others have speculated that an enhancement of mutability in the complementarity-determining regions (CDR; where mutations have a greater chance of being advantageous) and/or decrement of mutability in the framework regions (FR; where mutations are more likely to be lethal) may be accomplished by differential codon usage in concert with the known sequence specificity of the hypermutation mechanism. We have examined 115 nonproductively rearranged human Ig sequences. The mutation patterns in these unexpressed genes are unselected and therefore directly reflect inherent mutation biases. Using a chi2 test, we have shown that the number of mutations in the CDRs is significantly higher than the number of mutations found in the FRs, providing direct evidence for the hypothesis that mutations are preferentially targeted into the CDRs. PMID- 10368431 TI - Synonymous codon choices in the extremely GC-poor genome of Plasmodium falciparum: compositional constraints and translational selection. AB - We have analyzed the patterns of synonymous codon preferences of the nuclear genes of Plasmodium falciparum, a unicellular parasite characterized by an extremely GC-poor genome. When all genes are considered, codon usage is strongly biased toward A and T in third codon positions, as expected, but multivariate statistical analysis detects a major trend among genes. At one end genes display codon choices determined mainly by the extreme genome composition of this parasite, and very probably their expression level is low. At the other end a few genes exhibit an increased relative usage of a particular subset of codons, many of which are C-ending. Since the majority of these few genes is putatively highly expressed, we postulate that the increased C-ending codons are translationally optimal. In conclusion, while codon usage of the majority of P. falciparum genes is determined mainly by compositional constraints, a small number of genes exhibit translational selection. PMID- 10368432 TI - The nonrandom location of synonymous codons suggests that reading frame independent forces have patterned codon preferences. AB - Biased codon usage is common in eukaryotic and prokaryotic genes. Evidence from Escherichia, Saccharomyces, and Drosophila indicates that it favors translational efficiency and accuracy. However, to date no functional advantages have been identified in the codon-anticodon interactions involving the most frequently used (preferred) codons. Here we present evidence that forces not related to the individual codon-anticodon interaction may be involved in determining which synonymous codons are preferred or avoided. We show that the "off-frame" trinucleotide motif preferences inferrable from Drosophila coding regions are often in the same direction as Drosophila's "in-frame" codon preferences, i.e., its codon usage. The off-frame preferences were inferred from the nonrandomness of the location of confamilial synonymous codons along coding regions-a pattern often described as a context dependence of nucleotide choice at synonymous positions or as codon-pair bias. We relied on randomizations of the location of confamilial codons that do not alter, and cannot be influenced by, the encoded amino acid sequences, codon usage, or base composition of the genes examined. The statistically significant congruency of in-frame and off-frame trinucleotide preferences suggests that the same kind of reading-frame-independent force(s) may also influence synonymous codon choice. These forces may have produced biases in codon usage that then led to the evolution of the translational advantages of these motifs as preferred codons. Under this scenario, tRNA pool size differences between preferred and nonpreferred codons initially were evolved to track the default overrepresentation of codons with preferred motifs. The motif preference hypothesis can explain the structuring of codon preferences and the similarities in the codon usages of distantly related organisms. PMID- 10368433 TI - Current intraspecific dynamics of sequence evolution differs from long-term trends and can account for the AT-richness of honeybee mitochondrial DNA. AB - The very high AT content of hymenopteran mtDNA has warranted speculation about nucleotide substitution processes in this group. Here we investigate the pattern of honeybee, Apis mellifera, mtDNA nucleotide polymorphisms inferred from phylogeny in terms of differences between the ATPase6, COI, COII, COIII, cytochrome b, and ND2 genes and strand asymmetry in mutation rates. The observed transition/transversion ratios and the distribution of nonsynonymous substitutions between regions differed significantly. The pattern of differences between genes leading to these heterogeneities (the ATPase6 and COIII genes group apart from the rest) differed markedly from that predicted on the basis of long term evolutionary change and may indicate differences between current and long term dynamics of sequence evolution. Also, there is strong strand asymmetry in substitutions, which probably results in a mutability of G and C sufficiently high to account for the AT-richness of honeybee mtDNA. PMID- 10368434 TI - Two aspects of DNA base composition: G+C content and translation-coupled deviation from intra-strand rule of A = T and G = C. AB - The relative contribution of mutation and selection to the G+C content of DNA was analyzed in bacterial species having widely different G+C contents. The analysis used two methods that were developed previously. The first method was to plot the average G+C content of a set of nucleotides against the G+C content of the third codon position for each gene. This method was used to present the G+C distribution of the third codon position and to assess the relative neutrality of a set of nucleotides to that of the G+C content of the third codon position. The second method was to plot the intrastrand bias of the third codon position from Parity Rule 2 (PR2), where A = T and G = C. It was found that whereas intragenomic distributions of the DNA G+C content of these bacteria are narrow in the majority of species, in some species the G+C content of the minor class of genes distributes over wider ranges than the major class of genes. On the other hand, ubiquitous PR2 biases are amino acid specific and independent of the G+C content of DNA, so that when averaged over the amino acids, the biases are small and not correlated with the DNA G+C content. Therefore, translation coupled PR2 biases are unlikely to explain the wide range of G+C contents among different species. Considering all data available, it was concluded that the amino acid specific PR2 bias has only a minor effect, if any, on the average G+C content. In addition, PR2 bias patterns of different species show phylogenetic relationships, and the pattern can be as a taxal fingerprint. PMID- 10368436 TI - Estimating the contributions of selection and self-organization in RNA secondary structure. AB - In addition to characteristic structural properties imposed by evolutionary modification, evolved, single-stranded RNAs also display characteristic structural properties imposed by intrinsic physical constraints on RNA polymer folding. The balance of intrinsic and functionally selected characters in the folded conformation of evolved secondary structures was determined by comparing the predicted secondary structures of evolved and unevolved (random) RNA sequences. Though evolved conformations are significantly more ordered than conformations of random-sequence RNA, this analysis demonstrates that the majority of conformational order within evolved structures results not from evolutionary optimization but from constraints imposed by rules intrinsic to RNA polymer folding. PMID- 10368435 TI - The evolution of codon preferences in Drosophila: a maximum-likelihood approach to parameter estimation and hypothesis testing. AB - Synonymous codon usage in related species may differ as a result of variation in mutation biases, differences in the overall strength and efficiency of selection, and shifts in codon preference-the selective hierarchy of codons within and between amino acids. We have developed a maximum-likelihood method to employ explicit population genetic models to analyze the evolution of parameters determining codon usage. The method is applied to twofold degenerate amino acids in 50 orthologous genes from D. melanogaster and D. virilis. We find that D. virilis has significantly reduced selection on codon usage for all amino acids, but the data are incompatible with a simple model in which there is a single difference in the long-term Ne, or overall strength of selection, between the two species, indicating shifts in codon preference. The strength of selection acting on codon usage in D. melanogaster is estimated to be |Nes| approximately 0.4 for most CT-ending twofold degenerate amino acids, but 1.7 times greater for cysteine and 1.4 times greater for AG-ending codons. In D. virilis, the strength of selection acting on codon usage for most amino acids is only half that acting in D. melanogaster but is considerably greater than half for cysteine, perhaps indicating the dual selection pressures of translational efficiency and accuracy. Selection coefficients in orthologues are highly correlated (rho = 0.46), but a number of genes deviate significantly from this relationship. PMID- 10368437 TI - Coevolution of PERB11 (MIC) and HLA class I genes with HERV-16 and retroelements by extended genomic duplication. AB - The recent availability of genomic sequence information for the class I region of the MHC has provided an opportunity to examine the genomic organization of HLA class I (HLAcI) and PERB11/MIC genes with a view to explaining their evolution from the perspective of extended genomic duplications rather than by simple gene duplications and/or gene conversion events. Analysis of genomic sequence from two regions of the MHC (the alpha- and beta-blocks) revealed that at least 6 PERB11 and 14 HLAcI genes, pseudogenes, and gene fragments are contained within extended duplicated segments. Each segment was searched for the presence of shared (paralogous) retroelements by RepeatMasker in order to use them as markers of evolution, genetic rearrangements, and evidence of segmental duplications. Shared Alu elements and other retroelements allowed the duplicated segments to be classified into five distinct groups (A to E) that could be further distilled down to an ancient preduplication segment containing a HLA and PERB11 gene, an endogenous retrovirus (HERV-16), and distinctive retroelements. The breakpoints within and between the different HLAcI segments were found mainly within the PERB11 and HLA genes, HERV-16, and other retroelements, suggesting that the latter have played a major role in duplication and indel events leading to the present organization of PERB11 and HLAcI genes. On the basis of the features contained within the segments, a coevolutionary model premised on tandem duplication of single and multipartite genomic segments is proposed. The model is used to explain the origins and genomic organization of retroelements, HERV-16, DNA transposons, PERB11, and HLAcI genes as distinct segmental combinations within the alpha- and beta-blocks of the human MHC. PMID- 10368438 TI - Is there a phylogenetic signal in prokaryote proteins? AB - Using the sequence information from nine completely sequenced bacterial genomes, we extract 32 protein families that are thought to contain orthologous proteins from each genome. The alignments of these 32 families are used to construct a phylogeny with the neighbor-joining algorithm. This tree has several topological features that are different from the conventional phylogeny, yet it is highly reliable according to its bootstrap values. Upon closer study of the individual families used, it is clear that the strong phylogenetic signal comes from three families, at least two of which are good candidates for horizontal transfer. The tree from the remaining 29 families consists almost entirely of noise at the level of bacterial phylum divisions, indicating that, even with large amounts of data, it may not be possible to reconstruct the prokaryote phylogeny using standard sequence-based methods. PMID- 10368439 TI - A probable mixed-function supraoperon in Pseudomonas exhibits gene organization features of both intergenomic conservation and gene shuffling. AB - Sequencing of an 8182-bp chromosomal region in Pseudomonas stutzeri revealed the major portion of an apparent mixed-function supraoperon (defined as a nested organization of transcriptional units encoding gene products which function in more than one biochemical pathway). A nearly identical supraoperon organization was apparent in the unpublished Pseudomonas aeruginosa genome database, where the complete Pseudomonas supraoperon was deduced. The serC(pdxF)-aroQp. pheA-hisHb tyrAc-aroF-cmk-rpsA supraoperon encodes 3-phosphoserine aminotransferase, a bidomain chorismate mutase/prephenate dehydratase, imidazole acetol-phosphate aminotransferase, cyclohexadienyl dehydrogenase, 5-enolpyruvylshikimate 3 phosphate synthase, cytidylate kinase, and ribosomal protein S1. The member genes were identified by homology analysis, enzyme assay, and/or functional complementation. Although SerC(PdxF) and HisHb exercise their primary functions in serine, pyridoxine, and histidine biosynthesis, they also have critical catalytic roles in provision of the sidechain amino groups of tryptophan, phenylalanine, and tyrosine. The likelihood of supraoperon-wide translational coupling is suggested by the highly compressed intergenic spacing (including overlapping stop and start codons), as well as by possible hairpin structures in mRNA which may sequester some of the ribosome-binding sites and thus provide a mechanism for translational coupling. A comparison of the organization of the supraoperon genes in other organisms represented in the database revealed unmistakable conservation of the linkage of these genes across wide phylogenetic boundaries, albeit with considerable gene shuffling. At least remnants and shuffled portions of the entire supraoperon are distributed throughout the Gram negative bacteria with the hisHb-tyrA-aroF gene block being conserved as distantly as the gram-positive bacteria. Such conservation of mixed-function genes may reflect the selective value of still-unknown global relationships of protein-protein interaction or regulation. PMID- 10368440 TI - Organization, structure, and evolution of the nonadult rat beta-globin gene cluster. AB - The beta-globin gene cluster of Wistar rat was extensively cloned and the embryonic genes were mapped and sequenced. Four overlapping lambda Dash recombinant clones cover about 31 kb and contain four nonadult beta-globin genes, 5'-epsilon1-gamma1-gamma2-psigamma3-3'. The epsilon1 and gamma2 are active genes, since their protein products were detected in the fetal stage of the rat (Iwahara et al., J Biochem 119:360-366, 1996). The gamma1 locus might be a pseudogene, since the ATA box in the promoter region is mutated to GTA; however, no other defect is observed. The psigamma3 locus is a truncated pseudogene because a 19 base deletion, which causes a shift of the reading frame, is observed between the second nucleotide of the putative codon 68 and codon 76. A sequence comparison suggests that the psigamma3 might be produced by a gene conversion event of the proto-gamma-globin gene set. Possible histories of the evolution of rat nonadult beta-globin genes are discussed. PMID- 10368441 TI - Reconstructing the complex evolutionary history of hepatitis B virus. AB - A detailed analysis of the evolutionary history of hepatitis B virus (HBV) was undertaken using 39 mammalian hepadnaviruses for which complete genome sequences were available, including representatives of all six human genotypes, as well as a large sample of small S gene sequences. Phylogenetic trees of these data were ambiguous, supporting no single place of origin for HBV, and depended heavily on the underlying model of DNA substitution. In some instances genotype F, predominant in the Americas, was the first to diverge, suggesting that the virus arose in the New World. In other trees, however, sequences from genotype B, prevalent in East Asia, were the most divergent. An attempt was also made to determine the rate of nucleotide substitution in the C open reading frame and then to date the origin of HBV. However, no relationship between time and number of substitutions was found in two independent data sets, indicating that a reliable molecular clock does not exist for these data. Both the pattern and the rate of nucleotide substitution are therefore complex phenomena in HBV and hinder any attempt to reconstruct the past spread of this virus. PMID- 10368442 TI - Mitochondrial genes collectively suggest the paraphyly of Crustacea with respect to Insecta. AB - Complete sequences of seven protein coding genes from Penaeus notialis mitochondrial DNA were compared in base composition and codon usage with homologous genes from Artemia franciscana and four insects. The crustacean genes are significantly less A + T-rich than their counterpart in insects and the pattern of codon usage (ratio of G + C-rich versus A + T-rich codon) is less biased. A phylogenetic analysis using amino acid sequences of the seven corresponding polypeptides supports a sister-taxon status for mollusks-annelid and arthropods. Furthermore, a distance matrix-based tree and two most parsimonious trees both suggest that crustaceans are paraphyletic with respect to insects. This is also supported by the inclusion of Panulirus argus COII (complete) and COI and COIII (partial) sequence data. From analysis of single and combined genes to infer phylogenies, it is observed that obtained from single genes are not well supported in most topologies cases and notably differ from that of the tree based on all seven genes. PMID- 10368443 TI - Apolipophorin II/I, apolipoprotein B, vitellogenin, and microsomal triglyceride transfer protein genes are derived from a common ancestor. AB - Large lipid transfer proteins (LLTP) are nonexchangeable apolipoproteins and intracellular lipid-exchange proteins involved in the assembly, secretion, and metabolism of lipoproteins. We have identified contiguous conserved sequence motifs in alignments of insect apolipophorin II/I precursor (apoLp-II/I), human apolipoprotein B (apoB), invertebrate and vertebrate vitellogenins (VTG), and the large subunit of mammalian microsomal triglyceride transfer protein (MTP). Conserved motifs present in the N-terminal part of nonexchangeable apolipoproteins encompass almost completely the large subunit of MTP, suggesting a derivation from a common ancestral functional unit, termed large lipid transfer (LLT) module. Divergence of LLTP from a common ancestor is supported by (1) the statistical significance of the combined match scores obtained after motif-based database searches, (2) the presence of several identical amino acid residues in all LLTP sequences currently available, (3) the conservation of hydrophobic clusters in an alpha-helical domain, (4) the phylogenetic analysis of the conserved sequences related to the von Willebrand factor D (VWD) module identified in nonexchangeable apolipoproteins, and (5) the presence of four and one ancestral exon boundaries in the LLT and VWD modules, respectively. Our data indicate that the genes coding for apoLp-II/I, apoB, VTG, and the MTP large subunit are members of the same multigene superfamily. LLTP have emerged from an ancestral molecule designed to ensure a pivotal event in the intracellular and extracellular transfer of lipids and liposoluble substances. PMID- 10368444 TI - Circular permutations in the molecular evolution of DNA methyltransferases. AB - Circular permutations of genes during molecular evolution often are regarded as elusive, although a simple model can explain these rearrangements. The model assumes that first a gene duplication of the precursor gene occurs in such a way that both genes become fused in frame, leading to a tandem protein. After generation of a new start codon within the 5' part of the tandem gene and a stop at an equivalent position in the 3' part of the gene, a protein is encoded that represents a perfect circular permutation of the precursor gene product. The model is illustrated here by the molecular evolution of adenine-N6 DNA methyltransferases. beta- and gamma-type enzymes of this family can be interconverted by a single circular permutation event. Interestingly, tandem proteins, proposed as evolutionary intermediates during circular permutation, can be directly observed in the case of adenine methyltransferases, because some enzymes belonging to type IIS, like the FokI methyltransferase, are built up by two fused enzymes, both of which are active independently of each other. The mechanism for circular permutation illustrated here is very easy and applicable to every protein. Thus, circular permutation can be regarded as a normal process in molecular evolution and a changed order of conserved amino acid motifs should not be interpreted to argue against divergent evolution. PMID- 10368445 TI - Motion pictures in pediatric cardiology PMID- 10368447 TI - The athlete and heart disease: diagnosis, evaluation, and management PMID- 10368446 TI - Autonomic changes and heart rate variability in children with neurocardiac syncope. AB - This study evaluated resting autonomic function and autonomic responses to head up tilt-table testing in children who experienced neurocardiac syncope to determine whether predictable differences existed between these patients and normal volunteers. Neurocardiac syncope is a common cause of syncope in children. The mechanism, though related to abnormalities in autonomic function, has not been fully elucidated, particularly in pediatric patients. This study evaluated resting autonomic tone using noninvasive autonomic function tests (i.e., Valsalva, handgrip, and deep breathing) and 24-hour heart rate variability (HRV). In addition, heart rate and blood pressure were evaluated during head-up tilt examination. Values from patients who experienced neurocardiac syncope were compared to those from age-matched normal volunteers. No significant differences were noted during noninvasive testing. Some time domain HRV variables demonstrated a trend toward significant difference (p < 0.10). Tilt testing data were significantly different in sinus beat to sinus beat (RR) intervals between controls and syncope patients at 2, 5, and 10 minutes after tilting. In addition, significant differences were noted in RR interval and the standard deviation of RR interval 1 or 2 minutes prior to syncope when compared to controls at 5 and 10 minutes after tilting. Children with syncope exhibited abnormalities during tilt testing indicating an increased sympathetic or decreased parasympathetic tone, particularly prior to syncope. Some measures of HRV might constitute noninvasive parameters that correlate with the positive tilt table test. PMID- 10368448 TI - Effects of gamma-globulin on the cardiac sequelae of Kawasaki disease. AB - Our aim was to delineate the effect of various factors, such as sex, age, serum albumin levels, and the timing of gamma-globulin (GG) therapy, on cardiac sequelae of Kawasaki disease. The patients with Kawasaki disease who were reported at the 1995-1996 nationwide survey and received 2000 mg/kg at specified hospitals were selected as the subjects of the study. A total of 2221 patients actually received the basic dose. The relationships of the GG therapy with the cardiac sequelae, sex, age, timing of GG administration (the date of initiation and duration of the regimen following disease onset), and serum albumin levels were examined by using logistic regression analysis. The odds ratios for the cardiac sequelae in patients with Kawasaki disease were high in males (1.63), in those under the age of 1 year (1.54), and in those with a serum albumin level <3.2 g/dl (2.64). The odds ratio was low in those who received GG before day 8 of the illness (0.69) or in those for whom the administration period was for 2 days or less (0.67). To prevent cardiac sequelae of Kawasaki disease it is desirable that GG therapy be started as soon as possible and completed within 2 days. PMID- 10368449 TI - Left ventricular diastolic filling patterns associated with progressive anthracycline-induced myocardial damage: A prospective study. AB - The objective of this study was to examine changes in diastolic function associated with progressive myocardial damage and their implications. We used prospective sequential Doppler echocardiographic studies of left ventricular (LV) function. The study included 125 consecutive children (median age 6.3 years) receiving anthracyclines to cumulative doses between 45 and 1150 mg/m2 (median 270 mg/m2). We measured peak early (E) and atrial (A) phase filling velocities, EA ratio, deceleration and isovolumic relaxation times (EDecT and IVRT), heart rate, and fractional shortening (SF). Results were compared serially and with individually paired control data matched for body surface area. Progressive myocardial damage was evidenced by a mean SF decrease of 1 absolute %/100 mg/m2 of anthracycline. Six patients developed cardiac failure. After 1-100 mg/m2 of anthracyclines, the EA ratio decreased (mean 1.54-1.40, p = 0.02) and IVRT became prolonged (54 vs 52 msec in controls, p = 0.03). EA ratio increased again with the next dose, usually normalizing thereafter. Twelve patients ended treatment with an EA ratio <1 (1 cardiac death) and 17 with EA ratio >2 (2 cardiac deaths). Diastolic abnormalities were not strongly predictive of reduced SF. Modest changes in left ventricular diastolic filling patterns occur during anthracycline treatment of childhood malignancies. Although 20% of patients have significant abnormalities of diastolic filling by the end of treatment, considerable individual variability renders the pathophysiological and clinical implications of the early changes uncertain. PMID- 10368450 TI - Adult congenital heart disease on CD-ROM PMID- 10368452 TI - Technetium-99m hexamethyl propylenamine oxime lung clearance in the estimation of pulmonary hypertension in congenital heart disease: A preliminary comparative study with cardiac catheterization and pathology. AB - Thirty-six patients ranging in age from 7 months to 15 years and weighing from 5300 g to 49 kg (24 undergoing corrective surgery and 12 cases with reversed shunt and no operation) underwent technetium 99m hexamethyl propylenamine oxime (Tc-99m HMPAO) lung clearance study and the results were compared with catheterization and pathology. Patients were allocated into three groups with respect to pathological grading (Heath-Edwards' classification) and the results were correlated on the basis of pathology. In group I (grades I and II), Pearson correlation coefficient was 0.86 with pulmonary artery pressure (PAP), pulmonary vascular resistance (PVR), and Tc-99m HMPAO lung clearance (t1/2). Pearson correlation coefficients were 0. 863 and 0.88 in the second (grade III) and third group (with reversed shunt and no operation). There were statistically significant differences among the groups with respect to PAP, PVR, or t1/2. The results of radionuclide study (t1/2) were very well correlated within the groups with respect to hemodynamic parameters (PAP and PVR). Tc-99m HMPAO has potential as a highly sensitive indicator for detecting early and minimal microvascular lung injuries, and it may reflect accurate lung clearance and retention enabling an estimation of the state of pulmonary hypertension. PMID- 10368451 TI - Ultrasound myocardial tissue characterization by integrated backscatter in children treated with anthracyclines. AB - The objective of our study was to evaluate integrated backscatter (IBS) measurement, an ultrasound method of myocardial tissue characterization, in children receiving cardiotoxic anthracyclines for malignancy. Myocardial injury is known to diminish the normal cyclic variation of IBS (CVIBS) during the cardiac cycle. We used a cross-sectional, case-controlled study of children receiving anthracyclines and serial, prospective observation in a subgroup of children. The study took place in a university-affiliated, tertiary referral center for pediatric cardiology and oncology. Children undergoing routine echocardiograms before, during, and after anthracycline treatment participated in this study. Children evaluated in the cardiology clinic for innocent murmurs participated as controls. There was no intervention. CVIBS was measured using specialized echocardiographic software which quantitates the intensity of backscattered echoes returning from myocardial cells within a user-defined region of interest. Standard echocardiographic measures of left ventricular function were also made. The results indicated that abnormal CVIBS was prevalent during anthracycline treatment (17%) and at late follow-up (20%). In serial studies, CVIBS decreased in all children after anthracycline treatment. Anthracycline dose and time since last dose did not predict which children would have abnormalities of left ventricular function or of CVIBS. This report provides preliminary evidence that CVIBS may be a useful supplement to the noninvasive, echocardiographic assessment of the heart during anthracycline treatment in children. PMID- 10368453 TI - Inhalation of low-dose nitric oxide to evaluate pulmonary vascular reactivity in children with congenital heart disease. AB - The objective of this study was to investigate the efficacy of low-dose nitric oxide (NO). The study used fifteen consecutive Japanese preoperative patients (7 males and 8 females) with congenital heart disease and pulmonary hyptertension (mean pulmonary arterial pressure >30 mmHg), 6 of these patients had Down's syndrome. Hemodynamic measurements were taken in room air, 100% oxygen, 5 and 40 parts per million NO (NO5 and NO40) by inhalation. The differences between two observations within the same group were determined by the two-tailed paired t test. A pulmonary vascular resistance (Rp) regression curve was constructed by using linear regression analysis. The percentage change in pulmonary arterial pressure per systemic arterial pressure (Pp/Ps) with NO40 (Pp/Ps-40) exceeded that of Pp/Ps-5 (p < 0.0001). The percentage change for the Rp with NO40 (Rp-40) was larger than that for the Rp-5 (p = 0.0003). The percentage change of Pp/Ps-5 and that with oxygen were similar (p = 0.266). The relationship between Rp-5 and Rp-40 was linear. In conclusion, the effects of NO5 were equivalent to 100% oxygen but less than NO40. NO5 should initially be used to test pulmonary reactivity. If there is no response, patients should still be given NO40. PMID- 10368454 TI - Cardiac functions in children with vitamin D deficiency rickets. AB - Nutritional deficiency of vitamin D is common in developing countries as a result of both inadequate diet and exposure to ultraviolet light. The most striking biochemical finding in this illness is hypocalcemia. Reduction in serum calcium level may affect ventricular contraction. The purpose of this study was to evaluate prospectively left ventricular function in a group of 27 infants diagnosed as having rickets. Electrocardiograms and echocardiographic studies were undertaken in all patients. A group of ten healthy infants was used as a control for the echocardiographic examinations. Patients were divided into three groups according to the biochemical classification of rickets. There were eight patients in group I, nine in group II, and ten in group III. Abnormal electrocardiographic findings were noted in four infants in group I, three in group II, and six in group III before treatment of the rickets. These changes resolved following treatment. Echocardiographic studies revealed left ventricular dysfunction in the pretreatment stage. The most striking echocardiographic finding is the increase in the ratio of interventricular septal thickness to left ventricular posterior wall thickness in eight patients from group III. This returned to normal after treatment of the rickets. This study has demonstrated echocardiographic evidence of left ventricular dysfunction in children with rickets. These abnormalities were not, however, sufficiently severe to be associated with clinical signs of cardiac failure. Cardiomyopathy may develop in rickets, especially in the third stage of the disease, and this finding may return to normal following adequate treatment of the rickets. PMID- 10368455 TI - Antibody-mediated red blood cell agglutination resulting in spontaneous echocardiographic contrast. AB - Spontaneous echocardiographic contrast is well reported in states of low flow and low shear stress, and the primary blood component involved has been reported as red blood cells via rouleaux formation. This report describes the occurrence of spontaneous echocardiographic contrast from a unique mechanism of IgM-mediated red blood cell agglutination and describes the clinical sequelae. PMID- 10368456 TI - Noninvasive external pacing in the newborn. AB - Complete atrioventricular block in the newborn occasionally requires emergency intervention when the heart rate is too low to provide adequate cardiac output. While medications are frequently ineffective, emergent implantation of a pacemaker is not always feasible and carries a significant morbidity. We report two cases of complete atrioventricular block in newborns treated immediately after birth with an external noninvasive pacemaker system. PMID- 10368457 TI - Reversible cardiomyopathy secondary to alpha-interferon in an infant. AB - Interferon-alpha (IFN-alpha) is a biological response modifier with antiviral and tumoral effect that is used in the treatment of chronic myelogenous leukemias. Adverse effects are well documented and cardiovascular disturbances mostly include hypotension and tachycardia and rarely cardiomyopathy. We report on an infant with chronic myelomonocytic leukemia (CML) diagnosed at 3 months of age who was treated with increasing IFN-alpha dosage (2.5-5.5 million U/m2/day) given subcutaneously for 7.5 months. At that age, he presented anorexia, general malaise, and nocturnal sweating for about a week, followed by respiratory distress and tachycardia. Diagnosis of congestive heart failure was suspected and documented by cardiomegaly and echographic changes of left ventricular dilated cardiomyopathy, with a 40% left ventricular ejection fraction (EF) and 20% fractional shortening (FS). He was treated with digoxin, furosemide, and angiotensin converting inhibitors, and IFN-alpha was discontinued. Progressive improvement of cardiac function was observed within 7 months of the events with normalization of the echocardiographic findings (EF 60%, FS 31%). We should emphasize the possibility of severe and reversible cardiac toxicity of IFN-alpha in infancy. PMID- 10368458 TI - Cardiovascular involvement in a boy with Sweet's syndrome. AB - Acute febrile neutrophilic dermatosis (Sweet's syndrome) is a rare disease in infancy. It may present in an isolated manner or be associated with diverse conditions. Only two children with postinflammatory slack skin who developed cardiovascular disease have been described to date, both of whom died from coronary artery occlusion. We report a boy with Sweet's syndrome and diffuse vascular disease involving the aorta and the supraaortic vessels, the pulmonary trunk, and the right coronary artery but without signs of coronary obstruction. PMID- 10368459 TI - A case of anomalous origin of the pulmonary arteries: right pulmonary artery from the descending aorta and the left pulmonary artery from the ascending aorta. AB - A 3-month-old girl classified as having persistent truncus arteriosus underwent surgical correction of the anomalous origin of the pulmonary arteries; the right pulmonary artery from the descending aorta and the left pulmonary artery from the ascending aorta. The patient died on the fourth postoperative day. The definite diagnosis and choice of surgical strategies should be further examined. PMID- 10368460 TI - Post-Mustard procedure pulmonary venous obstruction: An opportunity for anatomic correction with a one-stage arterial switch. AB - A 14-year-old boy after a Mustard procedure for transposition of the great arteries developed pulmonary hypertension secondary to baffle obstruction. This occurred over several years without apparent significant symptomatology. Systemic level pressure prevailed in the left (pulmonary) ventricle and provided an opportunity to perform a successful one-stage arterial switch. PMID- 10368461 TI - Infantile hypertrophic cardiomyopathy of glycogenosis type IX: isolated cardiac phosphorylase kinase deficiency. AB - Glycogen storage disease confined to the heart due to cardiac phosphorylase kinase deficiency causes a fatal infantile cardiomyopathy. Cardiomegaly can be detected in utero and is progressive. Electrocardiographic and echocardiographic findings are characteristic but not specific; these include large QRS complexes, short PR interval, and a hypertrophic nonobstructive pattern. Conclusive diagnosis requires biochemical analysis of myocardium, which may not be possible premortem due to the amount of tissue required. Pathologic examination of a standard cardiac biopsy can provide a presumptive diagnosis. There is no current treatment except a heart transplant. Infants succumb to heart failure and/or respiratory compromise due to pulmonary compression. This is a rare entity; only three cases have been reported to our knowledge. We report two additional cases. PMID- 10368463 TI - From other journals PMID- 10368462 TI - Balloon disruption of pulmonary artery band in a child with congenital heart disease. PMID- 10368464 TI - Upcoming events in pediatric cardiology PMID- 10368465 TI - Jumping the broom toward eliminating health disparities. Presidential address. PMID- 10368466 TI - Risk factors for abruptio placentae and eclampsia: analysis of 445 consecutively managed women with severe preeclampsia and eclampsia. AB - OBJECTIVE: Our purpose was to characterize the clinical presentation or laboratory variables predictive of either abruptio placentae or eclampsia in women with severe preeclampsia. STUDY DESIGN: Prospective collection of perinatal data from 445 consecutively managed women with severe preeclampsia and eclampsia. Univariate analysis was used to determine which of the independent variables were significantly different between the groups (abruptio placentae vs no abruptio placentae; eclampsia vs no eclampsia). Those with significant differences were then entered into multiple logistic regression analysis to determine those characteristics that were independently related to the outcome variable (abruptio placentae or eclampsia). Before multivariate analysis, the independent variables with an interval scale of measurement were converted to a dichotomous scale, with the receiver-operator characteristic curve used to determine a cutoff level. RESULTS: Univariate analysis revealed statistical significance for the following variables associated with eclampsia: uric acid concentration, > 8.1 mg/dL; proteinuria (>3+); headache; visual symptoms; deep tendon reflexes >3+; serum albumin concentration, <3 mg/dL; and serum creatinine concentration, >1.3 mg/dL. However, with subsequent multivariate analysis, only headache and deep tendon reflexes >3+ remained significant. Univariate analysis for variables associated with abruptio placentae revealed an association between bleeding and platelet count <60,000/mm3. There was no association between abruptio placentae and eclampsia and systolic, diastolic, or mean arterial pressure, quantitative proteinuria, epigastric pain, bleeding, gestational age at delivery, history of preeclampsia, or chronic hypertension. CONCLUSION: Quantitative proteinuria and degree of blood pressure elevation were not predictive of either abruptio placentae or eclampsia, as has previously been suggested. The greatest morbidity associated with eclampsia occurred in women with preterm gestations not receiving medical attention. PMID- 10368467 TI - Antenatal testing among 1001 patients at high risk: the role of ultrasonographic estimate of amniotic fluid volume. AB - OBJECTIVE: Our goal was to compare the accuracy of the amniotic fluid index and the 2-diameter pocket technique with respect to accuracy in predicting an adverse pregnancy outcome among patients at high risk undergoing antenatal testing. STUDY DESIGN: All women with high-risk pregnancies and intact membranes who underwent antenatal testing during an 18-month period were prospectively enrolled. Ultrasonographic estimates of amniotic fluid volume were performed by means of the amniotic fluid index and the 2-diameter pocket technique. Relative risks with 95% confidence intervals and receiver operator characteristic curves were calculated for patients with an ultrasonographic estimate of oligohydramnios (amniotic fluid index of 5 cm or 2-diameter pocket of >15 cm2). Outcome variables studied were intrapartum and neonatal complications. RESULTS: Among 1001 patients the mean (+/-SD) amniotic fluid index was 10.5 +/- 5 cm and the mean (+/-SD) 2-diameter pocket was 18.7 +/- 13.6 cm2. Significantly more patients (46%) were considered to have oligohydramnios according to the 2 diameter pocket criteria than according to the amniotic fluid index (21%, P <.0001, relative risk 1.7, 95% confidence interval 1.5-1.8). No significant differences in the incidences of nonreactive nonstress test results, meconium stained amniotic fluid, cesarean delivery for fetal distress, low Apgar scores, or infants with cord pH of <7.10 were observed between the oligohydramnios and normal amniotic fluid groups (P >.05) when assessed by relative risk with confidence interval and by receiver operator characteristic curves. CONCLUSIONS: Current ultrasonographic measurements with the amniotic fluid index and the 2 diameter pocket technique are poor diagnostic tests to determine whether a patient is at high risk for an adverse perinatal outcome. PMID- 10368468 TI - Vaginal hysterectomy for the woman with a moderately enlarged uterus weighing 200 to 700 grams. AB - OBJECTIVES: The purpose of this study was to compare the surgical outcomes of women with moderately enlarged uteri undergoing vaginal hysterectomy with those of women with uteri of normal size undergoing vaginal hysterectomy. A secondary objective was to investigate the roles of uterine morcellation and laparoscopically assisted vaginal hysterectomy in the treatment of these women. STUDY DESIGN: Thirty consecutive women during a 2-year period with uterine enlargement to a weight of between 200 and 700 g underwent vaginal hysterectomy or laparoscopically assisted vaginal hysterectomy limited to lysis of adhesions or adnexectomy. These patients with uterine enlargement (group 1) were compared with 160 women with uteri weighing <200 g who also underwent vaginal hysterectomy or laparoscopically assisted vaginal hysterectomy during the same interval (group 2). The 2 groups were compared for total complications, operative time, hospital stay, perioperative hemoglobin concentration change, and use of vaginal uterine morcellation and laparoscopically assisted vaginal hysterectomy. RESULTS: Operative time for vaginal hysterectomy was significantly longer for women in group 1 than for women in group 2 (66.6 +/- 26.2 minutes vs 53.0 +/- 25.5 minutes, P =.008). There was a linear relationship between uterine weight and operative time: Operative time = 47.156 + 0.056 x Uterine weight (r = 0.20, F = 7.66, degrees of freedom 1, 188, P = .006). Vaginal morcellation of the uterus was needed in 80.0% of the women in group 1 and in 10.0% of the women in group 2 (P < .001). Two women in group 1 (6.7%) and 9 women (5.6%) in group 2 had laparoscopically assisted procedures for lysis of adhesions, adnexectomy, or both, unrelated to uterine size (P =.69). There were no significant differences between the 2 groups with respect to perioperative hemoglobin concentration change or hospital stay. Finally, the rates of major surgical complications were similar in the 2 groups (3.3% in group 1 vs 4.3% in group 2, P > .99, 95% confidence interval -8.1% to 5.9%). CONCLUSIONS: Although vaginal hysterectomy requires a modest increase in operative time, it is as safe and effective for the woman with a moderately enlarged uterus as for the woman with a uterus of normal size. Vaginal uterine morcellation is the key to a successful operation and obviates the need for either abdominal or laparoscopically assisted hysterectomy solely to deal with moderate uterine enlargement. PMID- 10368469 TI - Neonatal sepsis and death caused by resistant Escherichia coli: possible consequences of extended maternal ampicillin administration. AB - OBJECTIVE: Our goal was to evaluate the relationship between neonatal death caused by sepsis associated with ampicillin-resistant organisms and length of antibiotic exposure. STUDY DESIGN: All neonatal deaths from culture-positive sepsis over a 3-year period were examined. Infants who were delivered at either the University of Mississippi Medical Center or at Saint Barnabas Medical Center at >/=24 weeks' gestation and died within 7 days of life were included. Information on the organism causing sepsis and its sensitivities was collected, and the number of doses of ampicillin administered to the mother before delivery was determined. RESULTS: Of the 78 neonatal deaths, 35 met the inclusion criteria. There were 8 cases of sepsis from ampicillin-resistant Escherichia coli and 27 cases caused by other organisms. There was a statistically significant difference between the mean number of doses of ampicillin received by the ampicillin-resistant Escherichia coli group (17.6 +/- 5.5) compared with the other organisms group (4.9 +/- 3.6) (P <.001). CONCLUSION: A relationship exists between neonatal death caused by ampicillin-resistant Escherichia coli and prolonged antepartum exposure to ampicillin. PMID- 10368470 TI - Multiple courses of betamethasone to enhance fetal lung maturation do not suppress neonatal adrenal response. AB - OBJECTIVE: Our purpose was to evaluate the neonatal adrenal gland by provocative testing in neonates of mothers who had received multiple courses of betamethasone to enhance fetal lung maturity. STUDY DESIGN: Infants of mothers who had received >/=3 courses of betamethasone for fetal lung maturation were enrolled in the study. Twenty-four hours after delivery a baseline serum cortisol concentration was obtained. A synthetic adrenocorticotropic hormone (Cortrosyn) was administered (0.25 mg/1.73 m2). Two hours later a second serum cortisol concentration was obtained. An increase in serum cortisol in response to Cortrosyn was considered a positive test result. Nominal data were compared by means of the Student t test. RESULTS: There were 9 infants enrolled in the study. The mean number of betamethasone treatment cycles was 4.8 +/- 1.09. The mean baseline cortisol level was 2.23 +/- 0.52 microgram/dL, and the mean post adrenocorticotropic hormone cortisol level was 9.86 +/- 1.70 microgram/dL. All neonates had a positive adrenocorticotropic hormone test result. Stepwise linear regression showed no association between the number of courses of betamethasone treatment cycles and the post-adrenocorticotropic hormone cortisol concentration. CONCLUSION: Multiple weekly treatment cycles of betamethasone for fetal lung maturity administered between 24 and 34 weeks' gestation do not appear to cause adrenal suppression. PMID- 10368471 TI - Does an amniotic fluid index of /=34 weeks' gestation with an amniotic fluid index of 5 cm and the same pregnancy complication. Case patients were also matched with control subjects for maternal race, age, parity, and gestational age. RESULTS: Prospectively, 79 women at high risk with an amniotic fluid index of 5. PMID- 10368472 TI - Long-term outcome of nonconservative surgery (hysterectomy) for endometriosis associated pain in women <30 years old. AB - OBJECTIVE: This study was undertaken to evaluate the effect that a patient's age at the time of hysterectomy for endometriosis-associated pain has on long-term improvement in symptoms. STUDY DESIGN: An investigation of women who underwent hysterectomy for pelvic pain and endometriosis at <30 or >40 years of age was performed by means of medical records review and mailed questionnaires. Participants were asked to complete 2 standardized surveys, the Disruption of Functioning Index and the Beck Depression Inventory. RESULTS: Sixteen women in the study group (<30 years old) and 27 women in the control group returned completed questionnaires. Although similar proportions reported overall alleviation of pain, the study group was significantly more likely to report residual symptoms, such as dyspareunia and dysuria. This younger group also more often reported a sense of loss after hysterectomy and reported more overall disruption in different aspects of life. CONCLUSION: Women who undergo hysterectomy for pelvic pain and endometriosis at <30 years old are more likely than older women to have residual symptoms, to report a sense of loss, and to report more disruption from pain in different aspects of their lives. PMID- 10368473 TI - Effects of obstetrician characteristics on cesarean delivery rates. A community hospital experience. AB - OBJECTIVE: Despite a decrease in the overall cesarean delivery rate at Ravenswood Hospital Medical Center in Chicago, a wide range of variation existed among individual obstetricians' rates. This study evaluated obstetricians' characteristics to determine whether they affected cesarean delivery rates. STUDY DESIGN: In 1994 members of my department adopted strategies to decrease the cesarean delivery rate. Data on women who were delivered at the obstetric unit from 1994-1997 and data on their neonates were studied. Certain characteristics of obstetricians were also analyzed. The data were grouped according to personal characteristics and obstetricians' cesarean delivery rates: group 1 had a low rate (15%). Pearson chi2 analysis was used to evaluate the differences between the proportions. P <.05 was considered significant. RESULTS: The departmental cesarean delivery rate decreased from 20.5% in 1994 to 15.5% in 1997 (P <.0001), whereas individual obstetricians' rates varied from 0% to 44.4%. Obstetricians in group 1 (average rate 12.2%) and group 2 (average rate 20.8%, P <.0001) served similar populations with similar outcomes. Compared with obstetricians in group 2, those in group 1 (low rate) performed more vaginal deliveries after cesarean birth and used epidural analgesia and the vacuum extractor more frequently. Young age of physician, graduation from a domestic medical school, group practice, and smaller volume of births were all significantly linked to lower cesarean delivery rates. CONCLUSIONS: Cesarean delivery rates can safely be reduced. Certain individual obstetrician characteristics influence cesarean delivery rates. Obstetricians' commitment facilitates lowering of cesarean delivery rates. PMID- 10368474 TI - The spectrum of severe preeclampsia: comparative analysis by HELLP (hemolysis, elevated liver enzyme levels, and low platelet count) syndrome classification. AB - OBJECTIVE: This study was undertaken to explore the spectrum of maternal disease with a triple classification system of HELLP (hemolysis, elevated liver enzyme levels, and low platelet count) syndrome and compare these classes with severe preeclampsia without HELLP syndrome. STUDY DESIGN: In this retrospective analytic study the pregnancies of 777 patients with class 1, 2, or 3 HELLP syndrome were compared and contrasted with those of 193 women with severe preeclampsia but without HELLP syndrome. RESULTS: Eclampsia, epigastric pain, nausea and vomiting, significant proteinuria, major maternal morbidity, and stillbirth increased as HELLP syndrome worsened from class 3 to class 1. In contrast, headache and diastolic hypertension were more common among the significantly heavier patients with severe preeclampsia without HELLP syndrome. Approximately half of pregnancies complicated by class 1 HELLP syndrome exhibited significant maternal morbidity, compared with only 11% of those complicated by severe preeclampsia without HELLP syndrome. Although a significant trend was apparent in increasing levels of lactate dehydrogenase, aspartate aminotransferase, and uric acid as HELLP syndrome worsened, there was considerable variation within groups. CONCLUSION: Laboratory and clinical indices of disease severity in patients with severe preeclampsia or eclampsia generally were highest with class 1 HELLP syndrome and were lowest when HELLP syndrome was absent. Class 3 HELLP syndrome is considered a clinically significant transitional group. PMID- 10368475 TI - Planned cesarean hysterectomy: A preferred alternative to separate operations. AB - OBJECTIVE: The aim of this study was to evaluate our institutional experience with planned cesarean hysterectomy. STUDY DESIGN: In this retrospective case control investigation of a 16-year experience, 100 pregnant women who underwent planned cesarean hysterectomy were compared with 37 patients who underwent cesarean delivery followed by a hysterectomy performed within 6 months. RESULTS: Women undergoing planned cesarean hysterectomy did not have any demonstrable increase in intraoperative or postoperative complications when compared with the cesarean delivery plus later hysterectomy group. Primarily as a result of significantly reduced hospital stay and shorter total operative time, there was a significant financial advantage associated with a single planned cesarean hysterectomy with respect to separate operations. CONCLUSIONS: A policy to undertake planned cesarean hysterectomy for carefully selected patients appeared to produce advantages without increasing risks for these patients. Secondarily, it provided resident physicians the opportunity to learn the operation with supervision and under controlled circumstances. PMID- 10368476 TI - Steroid 5alpha-reductase 2 gene melting polymorphisms in male subjects with azoospermia or oligospermia. AB - OBJECTIVE: This study was designed to test the hypothesis that mutations in the gene for type 2 steroid 5alpha-reductase (SRD5A2) may be the cause of a phenotype characterized primarily by oligospermia or azoospermia. STUDY DESIGN: Deoxyribonucleic acid from control subjects and subjects with oligospermia (n = 12) and azoospermia (n = 6) were evaluated for mutations in SRD5A2. Methods used for mutation analysis included polymerase chain reaction, Southern blotting, and denaturing gradient gel electrophoresis. RESULTS: Denaturing gradient gel electrophoresis of all 5 amplified exons resulted in similar migration patterns in samples from both control and study subjects. Genomic deoxyribonucleic acid subjected to denaturing gradient gel electrophoresis after restriction digest revealed melting polymorphisms. Direct sequencing of the gene in a single patient with a unique melting polymorphism yielded a normal sequence. CONCLUSIONS: Melting polymorphisms for SRD5A2 were detected in a group of patients with oligospermia or azoospermia. Sequence analysis did not demonstrate functional mutations in the coding sequence of this gene. PMID- 10368477 TI - Immediate recall of oral contraceptive instructions: implications for providers. AB - OBJECTIVE: The object of the study was to determine the patient characteristics associated with inadequate recall of oral contraceptive pill-taking instructions. STUDY DESIGN: Sexually active women aged 13 to 40 years (n = 150) attending university-based family planning clinics completed anonymous self-report measures that assessed demographic and reproductive characteristics, understanding of pill taking instructions, and contraceptive compliance. Logistic regression was used to determine factors associated with inadequate recall for the sample, stratified by minority versus nonminority women. RESULTS: Minority women with inadequate recall were almost 6 times more likely than minority women with adequate recall not to know the name of the prescribed oral contraceptive and were 3 times more likely to have less than a high school education. In addition there were 1-fold and 2-fold increases in likelihood of inadequate recall as certainty of pill taking instructions and general oral contraceptive knowledge, respectively, decreased. Inadequate recall was associated with poor compliance. CONCLUSION: Women with inadequate recall may be identified at the conclusion of their visit so that interventions to enhance their pill-taking skills can be provided. PMID- 10368478 TI - Early risk assessment of severe preeclampsia: admission battery of symptoms and laboratory tests to predict likelihood of subsequent significant maternal morbidity. AB - OBJECTIVE: This study was undertaken to investigate the utility of an admission battery of findings and laboratory data in the discrimination of patients with severe preeclampsia with or without HELLP (hemolysis, elevated liver enzyme levels, and low platelet count) syndrome at high risk for development of significant maternal morbidity. STUDY DESIGN: The clinical and laboratory findings at hospital admission for 970 patients with severe preeclampsia with or without HELLP syndrome were studied retrospectively to develop parameters associated with low, moderate, and high risks for the subsequent development of significant maternal morbidity involving the hematologic and coagulation, cardiopulmonary, and hepatorenal systems. RESULTS: Nausea and vomiting and epigastric pain are independent risk factors for complicated severe preeclampsia. Results of a panel of tests with values including lactate dehydrogenase level >1400 IU/L, aspartate aminotransferase level >150 IU/L, alanine aminotransferase level >100 IU/L, uric acid level >7.8 mg/dL, serum creatinine level >1.0 mg/dL, and 4+ urinary protein by dipstick can be used to discriminate the patient at high risk for significant maternal morbidity. Concentrations of lactate dehydrogenase, aspartate aminotransferase, and uric acid above these cut points have the strongest predictive value and are risk additive with worsening thrombocytopenia. CONCLUSION: The presence of nausea and vomiting, epigastric pain, or both in association with admission laboratory values that are in excess of the cutoffs for lactate dehydrogenase, aspartate aminotransferase, and uric acid concentrations or for all 6 tests is predictive of high risk of morbidity for the patient with severe preeclampsia. These factors are independent of and additive with the rising maternal risk associated with decreasing platelet count. PMID- 10368479 TI - A comparison of women with primary and recurrent pelvic prolapse. AB - OBJECTIVE: Our purpose was to identify clinically relevant differences in women with primary and recurrent pelvic organ prolapse. STUDY DESIGN: Consecutive women undergoing reconstructive surgery completed a urogynecologic history and physical examination and underwent either multichannel urodynamic testing or pelvic floor fluoroscopy, or both. Two groups were compared: primary (no prior surgery for pelvic organ prolapse) and recurrent. RESULTS: One hundred eighty-one consecutive women were studied-103 with primary and 78 with recurrent prolapse. The groups were similar with respect to age, race, weight, vaginal parity, prolapse stage, urodynamic diagnosis, extent of visceral malposition, and common urinary, anorectal, and sexual symptoms. Clinically relevant differences were found, with the recurrent group having shorter vaginal lengths (P =. 0005), being more likely to have had a hysterectomy for a nonprolapse indication (P =.00018) and to be receiving hormone replacement therapy (P =.00003). CONCLUSION: The women with primary and recurrent pelvic organ prolapse in this population were remarkably similar in many quantifiable parameters measured. The clinical differences may be related to previous surgery for pelvic organ prolapse. PMID- 10368480 TI - Calciuria in symptom-free primigravid women remote from term: is the response to an oral calcium challenge predictable? AB - OBJECTIVE: This study was undertaken to compare the calciuric response in symptom free primigravid women to an oral calcium load between those with normal urinary calcium excretions and those with relatively low urinary calcium excretions. STUDY DESIGN: This was a prospective clinical trial. Eligible primigravid women between 16 and 20 weeks' gestation provided a 24-hour urine sample for determination of urinary calcium/urinary creatinine ratio. On the basis of these results the patients were divided into 2 groups: a relatively hypocalciuric group, in which the urinary calcium excretion was 3.4 mg. kg-1. 24 h-1. All participants undertook a 3-day low calcium dietary regimen. On the fourth day women underwent an oral calcium challenge. A 2-hour urine sample was collected before ingestion of 1 g calcium carbonate (preload). One hour after ingestion the women again collected a 2-hour urine sample (postload). The urinary calcium/urinary creatinine ratios in the preload and postload samples were determined and compared within and between the groups. RESULTS: The mean change (+/-SD) between the preload and postload urinary calcium/urinary creatinine ratios in the relatively hypocalciuric group was 0.60 +/- 1.44 (P =.04); that in the normocalciuric group was 3.09 +/- 2.26 (P =.11 ). There was a 5-fold difference in the response to calcium load between the hypocalciuric women and the normocalciuric women (0.60 vs 3.09), although this difference was not statistically significant (P =.20). CONCLUSIONS: Both hypocalciuric and normocalciuric women responded to an oral calcium challenge by an increase in the calcium excretion. The cause of the hypocalciuria in women at increased risk for preeclampsia is therefore not simply poor absorption of calcium. PMID- 10368481 TI - Brain stem auditory evoked potentials in the human fetus during labor. AB - OBJECTIVE: The aim of this study was to record, during labor, the brain stem auditory evoked potentials of the fetus from standard fetal scalp electrodes. STUDY DESIGN: A personal computer-based instrument was developed to record, during labor, brain stem auditory evoked potentials from 10 fetuses ranging in gestational age from 36 to >/=40 weeks. Auditory stimulus was provided by clicks (16/s) delivered on the mother's abdomen with an intensity of 120 dB. The evoked potential signals were digitized and averaged. Interfering artifacts were excluded from the averages. RESULTS: In 80% of the subjects, all the principal waveforms of the brain stem auditory evoked potentials-waves I, III, and V-were clearly identifiable. The latency for wave I ranged from 1.2 to 2.2 ms, wave III from 3.4 to 5.6 ms, and wave V from 5.8 to 8.4 ms. The morphologic features of the waveform and the interwave latency values were similar to those of normal term infants recorded in the past studies. CONCLUSION: We have reestablished that it is possible to record auditory evoked potentials during labor from fetal scalp electrodes. Brain stem auditory evoked potentials may be a useful screening tool for at least the severely neurologically damaged fetus. PMID- 10368482 TI - A study of antenatal cocaine use-chaos in action. AB - OBJECTIVE: This study identified behaviors or conditions associated with cocaine use among prenatal patients and evaluated pregnancy outcomes. STUDY DESIGN: A case-control study of patients attending a neighborhood-based prenatal program was conducted. For each patient who used cocaine, the next prenatal registrant with both a negative history of cocaine use and a negative urine screen for cocaine served as the control. RESULTS: Cocaine use was associated with older, multiparous women who had a history of prior low birth weight infants. Prenatal care was obtained later and less frequently. Other substances, including tobacco, alcohol, and marijuana, were more commonly used. A history of physical abuse and violence, as well as incarceration, was identified more often. The prevalence of syphilis was higher. Women who used cocaine were delivered of their infants earlier; prematurity occurred more often. Their infants were smaller. Regression analysis identified factors other than cocaine as important in either low birth weight or preterm delivery. Attainment of a greater number of prenatal care visits was associated with improved outcome. CONCLUSION: Women who use cocaine have numerous significant life disturbances, which may collectively influence pregnancy outcome. Cocaine use itself is a marker and did not appear to influence the prevalence of low birth weight or preterm delivery. PMID- 10368483 TI - Placenta previa: a 22-year analysis. AB - OBJECTIVE: Our purpose was to identify what anesthetic method is safer for women with a placenta previa. STUDY DESIGN: We retrospectively reviewed all women with placenta previa who underwent cesarean delivery during the period January 1, 1976 December 31, 1997 at Northwestern Memorial Hospital. RESULTS: Of 93,384 deliveries, placenta previa was found in 514 women. Identifiable trends with time included an increasing incidence of placenta previa (r = 0.54, P <.01); cesarean hysterectomy (r = 0.54, P <.01); placenta accreta (r = 0.45, P <.03); and regional anesthesia (r = 0.84, P <.0001). The mean gestational age at delivery was 35.3 +/- 3.4 weeks and did not change with time. General anesthesia was used for delivery in 380 women and regional anesthesia was used for 134 women. Prior cesarean delivery and general anesthesia were independent predictors of the need for blood transfusion, but only prior cesarean delivery was a predictor of the need for hysterectomy. General anesthesia increased the estimated blood loss, was associated with a lower postoperative hemoglobin concentration, and increased the need for blood transfusion. Elective and emergent deliveries did not differ in estimated blood loss, in postoperative hemoglobin concentrations, or in the incidence of intraoperative and anesthesia complications. Regional and general anesthesia did not differ in the incidence of intraoperative and anesthesia complications. CONCLUSIONS: In women with placenta previa, general anesthesia increased intraoperative blood loss and the need for blood transfusion. Regional anesthesia appears to be a safe alternative. PMID- 10368484 TI - Evaluation of fetal lung maturity in diamniotic twins. AB - OBJECTIVE: This study was undertaken to evaluate the correspondence in fetal lung development between diamniotic twins. STUDY DESIGN: Lecithin/sphingomyelin ratios were determined for amniotic fluid specimens collected from each sac in 58 diamniotic twin pregnancies. RESULTS: Overall, the lecithin/sphingomyelin ratios of twin A and twin B and those of the larger and smaller twins of each pair were closely correlated (r >/= 0.83, P <.001). When stratified by gestational age, however, the percentage disparity in lecithin/sphingomyelin ratios between members of twin pairs was significantly greater at 32 weeks' gestation (15%, P =.027). An analysis of the lecithin/sphingomyelin ratios of twins with a large lecithin/sphingomyelin ratio disparity (>/=20%) suggested that this disparity was a result of 1 twin having a lecithin/sphingomyelin ratio that was advanced for gestational age with respect to that of the co-twin. Disparities in lecithin/sphingomyelin ratio were not significantly affected by fetal sex or by discordance in size between the twins. CONCLUSION: At relatively early stages of diamniotic twin gestation (90%. An additional benefit of the latter procedure is that fertility is maintained. We report our experience at Stanford University Medical Center in which this technique was used in 6 cases within the past 5 years. STUDY DESIGN: Six women between the ages of 18 and 41 years underwent placement of arterial catheters for emergency (n = 3) or prophylactic (n = 3) control of postpartum bleeding. Specific diagnoses included cervical pregnancy (n = 1), uterine atony (n = 3), and placenta previa and accreta (n = 2). RESULTS: Control of severe or anticipated postpartum hemorrhage was obtained with transcatheter embolization in 4 patients. A fifth patient had balloon occlusion of the uterine artery performed prophylactically, but embolization was not necessary. In a sixth case, bleeding could not be controlled in time, and hysterectomy was performed. The only complication observed with this technique was postpartum fever in 1 patient, which was treated with antibiotics and resolved within 7 days. CONCLUSIONS: Uterine artery embolization is a superior first-line alternative to surgery for control of obstetric hemorrhage. Use of transcatheter occlusion balloons before embolization allows timely control of bleeding and permits complete embolization of the uterine arteries and hemostasis. Given the improved ultrasonography techniques, diagnosis of some potential high-risk conditions for postpartum hemorrhage, such as placenta previa or accreta, can be made prenatally. The patient can then be prepared with prophylactic placement of arterial catheters, and rapid occlusion of these vessels can be achieved if necessary. PMID- 10368489 TI - Perinatal risks associated with borderline amniotic fluid index. AB - OBJECTIVE: Our purpose was to determine whether a borderline amniotic fluid index observed during antepartum testing confers a significant risk of adverse perinatal outcome. STUDY DESIGN: We conducted a retrospective review of all patients entering antepartum testing at Los Angeles County-University of Southern California Women's and Children's Hospital during a 4-month period beginning January 1, 1996. Women with singleton pregnancies who underwent antepartum testing within 1 week of delivery and who were delivered at our institution were identified for our study. An amniotic fluid index >5 and <10 cm was defined as "borderline" and an amniotic fluid index of 10 to 24 cm was considered normal. Markers of adverse perinatal outcome included intrapartum fetal distress, 5 minute Apgar score <7, meconium-stained amniotic fluid, and intrauterine growth restriction. RESULTS: There was a 2-fold increase in the incidence of adverse perinatal outcome among the women with borderline amniotic fluid index in comparison with control subjects with normal amniotic fluid volume. This difference reflected a 4-fold increase in the incidence of fetal growth restriction among women with a borderline amniotic fluid index. CONCLUSIONS: A borderline amniotic fluid index observed in antepartum testing is associated with an increased risk of intrauterine growth restriction and overall adverse perinatal outcome. These observations suggest that borderline amniotic fluid index merits twice-weekly antepartum testing. PMID- 10368490 TI - Young age as a prognostic factor in cervical cancer: results of a population based study. AB - OBJECTIVE: Our goal was to use population-based data to determine the difference in 5-year survival in women diagnosed with cervical cancer between those aged 18 34 years and those aged 40-60 years. STUDY DESIGN: The SEER (Surveillance, Epidemiology, and End Results) public-use database, 1973-1994, was used for this investigation. Only subjects with cervical carcinoma diagnosed between 1988 and 1990 were included. Subjects were stratified on age at diagnosis (<35 years or 40 60 years), clinical stage, histologic type, race-ethnicity, and grade. RESULTS: Two thousand cases of invasive cervical cancer were identified. The younger subgroup of patients was diagnosed with earlier-stage disease more frequently than the older group (P =.0001). When adjustments were made for non-cervical cancer causes of death, there was no difference in 5-year survival between the 2 cohorts. African American women had a poorer 5-year survival (P =.02) CONCLUSION: There was no overall difference in survival between the 2 cohorts when appropriate adjustments were made for cause of death and for stage, histologic type, and grade of disease. PMID- 10368491 TI - Isolated pleural effusion in severe ovarian hyperstimulation: A case report. AB - Assisted reproductive technology programs use controlled ovarian hyperstimulation to maximize pregnancy rates. Severe ovarian hyperstimulation syndrome is a well known risk. Pleural effusion often accompanies severe ovarian hyperstimulation syndrome. We describe 2 cases of isolated hydrothorax without concomitant ascites and review the literature of this rare finding. PMID- 10368492 TI - Initial experience with extended culture and blastocyst transfer of cryopreserved embryos. AB - OBJECTIVE: Our purpose was to evaluate the viability and transfer efficiency of cryopreserved embryos allowed to develop into blastocysts in extended culture for in vitro fertilization. STUDY DESIGN: The embryos for in vitro fertilization that had been cryopreserved at either 2 PN (pronuclear) or cleaving stage (day 1-3) were thawed and cultured for uterine transfer on day 5. Outcome for day 5 embryo transfer was prospectively compared with previous outcomes from embryos transferred on day 2 or 3. RESULTS: For embryos thawed and transferred on day 2 or 3 (n = 99), the pregnancy rate was 33%, the implantation rate per embryo transferred was 15.2%, and the rate of multiple gestations was 42.4% (14/33) with 35.7% of pregnancies having >/=3 gestational sacs. For extended culture embryos transferred on day 5 (n = 25), the pregnancy rate was 36%, the implantation rate per embryo transferred was 16.7%, and the rate of multiple gestations was 33.3% (3/9) with all of these being twins. For embryo transfers performed on day 5 in which only blastocysts were transferred (n = 9), the pregnancy rate was 66.7%, the implantation rate per blastocyst was 44.4% (greater than the rate for the day 2 or 3 embryos, P =.0043), and the rate of multiple gestations was 33.3% (2/6) with all of these being twins. In extended culture 29.8% of cryopreserved embryos progressed to the blastocyst stage. In this series 4 subjects (15.4%) did not have blastocysts by day 5. CONCLUSION: Acceptable pregnancy rates can be obtained from cryopreserved embryos cultured to the blastocyst stage with a significantly higher implantation rate. Transfer of embryos that have "self-selected" to blastocysts results in reduced risk of higher-order (>2) multiple gestations, because only 1 or 2 embryos are transferred. PMID- 10368493 TI - Effect of magnesium prophylaxis and preeclampsia on the duration of labor. AB - OBJECTIVE: Our goals were to compare duration of labor at term for (1) women with preeclampsia versus normotensive nulliparous women and (2) nulliparous women with preeclampsia who received magnesium for seizure prophylaxis versus those who did not. STUDY DESIGN: We performed a retrospective cohort study of all nulliparous, term vaginal deliveries from 1989 through 1995 at University of California, San Francisco. The perinatal database and medical records were reviewed for information on duration of labor, maternal and labor characteristics, and neonatal outcomes. The chi2 odds ratio, and Student t tests were used to compare categoric and continuous variables between women with preeclampsia and control women and between women with preeclampsia who did and those who did not receive magnesium. Logistic regression was used to evaluate variables predictive of labor duration. RESULTS: Our study subjects were 4083 normotensive nulliparous women and 154 women with preeclampsia. A sample size calculation revealed that 1764 normotensive control subjects were needed to show a 10% difference in labor duration with 80% power and alpha of 0.05. Among women with preeclampsia, 93 (60%) were treated with magnesium and 61 (40%) were not. More women with preeclampsia than normotensive women had induction of labor and received epidural anesthesia, prostaglandin gel, and oxytocin (P <.003). Total labor duration did not differ between women with preeclampsia and normotensive women (P =.15) or between women with preeclampsia who received magnesium and those who did not (P =.09). In comparison with normotensive women, those with preeclampsia had a higher rate of postpartum hemorrhage (31% vs 22%, P =.005), and the rate was even higher among preeclamptic women treated with magnesium versus those who received no magnesium (34% vs 26%, P =.002). Logistic regression, with prolonged first stage of labor (>12 hours) used as the outcome variable, indicated that epidural anesthesia (odds ratio 2.3, 95% confidence interval 1.9-2. 6), oxytocin (odds ratio 1.8, 95% confidence interval 1.6-2.2), and persistent occipitoposterior presentation (odds ratio 1.6, 95% confidence interval 1.1-2.4) were associated with prolonged labor, whereas preeclampsia (odds ratio 0.9, 95% confidence interval 0.7-1. 1) and treatment with magnesium were not (odds ratio 1.1, 95% confidence interval 0.9-1.4). Induction (odds ratio 0.5, 95% confidence interval 0.4-0.6) and birth weight <2500 g (odds ratio 0. 5, 95% confidence interval 0.4 0.8) were associated with faster labor. CONCLUSIONS: In term nulliparous women, neither preeclampsia nor magnesium prophylaxis affected labor duration. PMID- 10368494 TI - Serum and tissue hormone levels of vaginally and orally administered estradiol. AB - OBJECTIVE: Our purpose was to determine serum and endometrial estradiol levels when micronized estradiol is administered vaginally and orally. STUDY DESIGN: Five subjects were given oral estradiol (2 mg twice daily), during an artificial luteal phase, and another group of 5 subjects were given the same dose of estradiol by the vaginal route. Endometrial biopsies and blood samples were obtained on day 21 of the cycle, 2 hours after the last dose was administered. Tissue and blood samples were assayed for estradiol. RESULTS: Serum estradiol levels were significantly higher with vaginally administered estradiol than with orally administered estradiol (2344 +/- 398 vs 279 +/- 76 pg/mL, P <.005). Endometrial estradiol concentrations were also significantly higher with vaginally administered estradiol than with the oral preparation (918 +/- 412 vs 13 +/- 2 pg/mg protein, P <.05). CONCLUSIONS: Vaginal administration of estradiol is more effective in increasing serum and endometrial levels of estradiol than the oral route and may represent the optimal route of administration for recipients of egg donation. If the vaginal route of estradiol administration is considered for menopausal replacement therapy, much lower doses of the standard oral quantities should be used. Furthermore, if the uterus is present, a progestin must be used to compensate for the high tissue levels of estradiol. PMID- 10368495 TI - Clinical value of mammography for symptomatic women 35 years of age and younger. AB - OBJECTIVE: This retrospective observational study was designed to answer the following question: Is mammography clinically effective in the evaluation of women 3 years. A partial ablation is defined as ablation of only the anterior or posterior endometrial wall and avoidance of the cornual areas. A posterior myometrial biopsy to determine the depth of adenomyosis was performed and correlated with outcome. The intrauterine cavity was evaluated postoperatively for adhesions. RESULTS: Patients without deep adenomyosis had good outcome after partial ablation. No postoperative intrauterine adhesions were found. CONCLUSION: Partial endometrial ablation can successfully treat menorrhagia in patients without deep adenomyosis. It does not cause intrauterine adhesions, which may lead to hematometra and potentially delay the diagnosis of endometrial cancer. PMID- 10368500 TI - Determinants of the outcome of intrauterine insemination: analysis of outcomes of 9963 consecutive cycles. AB - OBJECTIVE: Our aim was to determine which factors influence the effectiveness of intrauterine insemination. STUDY DESIGN: This article is a retrospective statistical analysis of outcomes of 9963 consecutive intrauterine insemination cycles. RESULTS: Patient age was the main determinant of pregnancy outcome (analysis of variance F ratio = 29, P <.0001), followed by the number of follicles at the time of intrauterine insemination (analysis of variance F ratio = 9, P <.0001) and sperm motility in the inseminate (analysis of variance F ratio = 4, P =.002). A total of 18.9% of all patients <26 years old conceived, compared with 13.9% of those 26-30 years old, 12.4% of those 31-35 years old, 11.1% of those 36-40 years old, 4.7% of those 41-45 years old, and 0.5% of patients >45 years old (P <.001). When analyzed by single years, ongoing pregnancy rates after intrauterine insemination remained high through age 32 years. Across all ages and causes of infertility, 7.6% of patients with 1 follicle at the time of intrauterine insemination conceived, compared with 10. 1% with 2, 14.0% with 4, and 16.9% with 6 follicles (P <.01). When ovulation occurred before intrauterine insemination (ie, no visible follicular structures), 4.6% of patients conceived. The likelihood of pregnancy was maximized when motile sperm numbers were >/=4 million and sperm motility was >/=60%. Differences in pregnancy outcomes between sperm processing options were related to differences in sperm motility after processing; use of methods incorporating motility enhancement with pentoxifylline and motile sperm concentration through silica gradients yielded the highest overall pregnancy rates. CONCLUSION: When the results of ongoing retrospective analysis of intrauterine insemination outcomes are applied, overall intrauterine insemination pregnancy rates have increased from 5.8% per cycle in 1991 to 13.4% per cycle in 1996, during which time the average age of patients undergoing intrauterine insemination has increased from 36.1 (+/-0.2) to 39.2 (+/-0.1) years. PMID- 10368501 TI - Uterine rupture associated with the use of misoprostol in the gravid patient with a previous cesarean section. AB - OBJECTIVE: Our purpose is to report our experience with uterine rupture in patients undergoing a trial of labor after previous cesarean delivery in which labor was induced with misoprostol. The literature on the use of misoprostol in the setting of previous cesarean section is reviewed. STUDY DESIGN: This report was based on case reports, a computerized search of medical records, and literature review. RESULTS: Uterine rupture occurred in 5 of 89 patients with previous cesarean delivery who had labor induced with misoprostol. The uterine rupture rate for patients attempting vaginal birth after cesarean section was significantly higher in those who received misoprostol, 5.6%, than in those who did not, 0.2% (1/423, P =.0001). Review of the literature reveals insufficient data to support the use of misoprostol in the patient with a previous cesarean delivery. CONCLUSION: Misoprostol may increase the risk of uterine rupture in the patient with a scarred uterus. Carefully controlled studies of the risks and benefits of misoprostol are necessary before its widespread use in this setting. PMID- 10368502 TI - Misoprostol is more efficacious for labor induction than prostaglandin E2, but is it associated with more risk? AB - OBJECTIVE: Our purpose was to compare the efficacy and safety of misoprostol with dinoprostone (Prepidil) for labor induction. STUDY DESIGN: In a randomized, controlled trial of labor induction, patients were randomly assigned to receive either 50 microgram of intravaginal misoprostol every 4 hours or 0.5 mg of intracervical prostaglandin E2 every 6 hours. Eligibility criteria included gestation of >/=31 weeks, Bishop score <6, and fewer than 12 contractions per hour. Primary outcomes were cesarean section, induction to delivery time, oxytocin use, and fetal distress requiring delivery. RESULTS: One hundred fifty nine women were randomly assigned to receive misoprostol (n = 81) or Prepidil (n = 78). There were no differences in the indication for induction, preinduction Bishop score, epidural use, or cesarean section rate. Mean time to delivery was significantly shorter in the misoprostol group (19 hours 50 minutes) than in the Prepidil group (28 hours 52 minutes) (P =.005). Only 58% of women in the misoprostol group required oxytocin augmentation, in comparison with 88% of women receiving Prepidil (P =.00002). However, 41% of women receiving misoprostol and 17% receiving Prepidil had late decelerations or bradycardias (P =.001), and 20% of the misoprostol group and 5% of the Prepidil group had deliveries for fetal distress (P =.05). CONCLUSIONS: Misoprostol is more efficacious than Prepidil for labor induction. However, the significantly increased incidence of abnormal fetal heart rate tracings and the trend in increased deliveries for fetal distress with misoprostol dosing of 50 microgram every 4 hours are of concern. These data suggest that either a lower dose of misoprostol or less frequent dosing of misoprostol should be considered. PMID- 10368503 TI - Comparison of the safety and efficacy of intravaginal misoprostol (prostaglandin E1) with those of dinoprostone (prostaglandin E2) for cervical ripening and induction of labor in a community hospital. AB - OBJECTIVE: This clinical trial evaluated the efficacy of intravaginal misoprostol (prostaglandin E1) and compared it with that of dinoprostone (prostaglandin E2) for cervical ripening and induction of labor in a community hospital. STUDY DESIGN: This study involved a retrospective analysis of 81 patients undergoing cervical ripening and induction of labor with prostaglandin E2 from May 1, 1996, to May 1, 1997. A comparison prospective analysis of 145 patients undergoing the same procedure with prostaglandin E1 from May 1, 1997 to May 1, 1998, was performed. RESULTS: The mean time to delivery was significantly shorter with misoprostol (19.8 +/- 10.4 hours) than with prostaglandin E2 (31.3 +/- 13.0 hours, P <.001). Delivery within 24 hours of induction was significantly more frequent with misoprostol (71.9% of subjects vs 31.3%, P <.001). There was no difference in the cesarean delivery rate with misoprostol (25.6% vs 22.2%, P <.67). The incidence of uterine hyperstimulation was higher with prostaglandin E2 (7.4% vs 0.7%, P <.007). There were no uterine ruptures with prostaglandin E2. There were 2 uterine ruptures and 1 dehiscence with prostaglandin E1 in 3 patients with previous cesarean deliveries and 1 rupture in a patient without a history of uterine scarring. There was no difference in neonatal outcome, with the exception of a fetal death related to uterine rupture in the misoprostol group. CONCLUSIONS: Compared with prostaglandin E2, misoprostol is more effective in cervical ripening and induction of labor, is as safe for patients who do not have a history of cesarean birth, may carry a higher incidence of uterine rupture, and should not be used for patients attempting vaginal birth after previous cesarean delivery. PMID- 10368505 TI - Is tocolysis safe in the management of third-trimester bleeding? AB - OBJECTIVE: Expectant management is among the current treatment options for pregnancies complicated by third-trimester bleeding at <36 weeks' gestation. The use of tocolytic agents to stop associated contractions is still somewhat controversial, however, and the number of cases reported to date is small. The purpose of our study was to find a large number of cases of preterm third trimester bleeding that was treated with tocolytic agents and evaluate them for any evidence of potential harm related to the use of these agents. STUDY DESIGN: Every case of third-trimester bleeding for a 6-year period was obtained from a perinatal database that was created as patients were hospitalized. Only cases of patients with onset of bleeding between 23 and 36 weeks' gestation were analyzed. Data collected included the gestational age at the time of first bleeding, the gestational age at delivery, whether tocolytic agents were used, the need for transfusion, maternal morbidity, and neonatal outcome. RESULTS: A total of 236 cases, consisting of 131 cases of abruptio placentae and 105 cases of placenta previa, met the study criteria. In the abruptio placentae group 95 women (73%) were treated with tocolytic agents. In this group the mean gestational age at the time of first bleeding was 28.9 weeks, the mean time from bleeding until delivery was 18.9 days, the median time from bleeding until delivery was 7 days, and the neonatal mortality rate was 51 deaths/1000 live births. In the placenta previa group 76 patients (72%) were treated with tocolytic agents. In this group the mean gestational age at first bleeding was 29.5 weeks, the mean time from bleeding until delivery was 29.3 days, the median time from bleeding until delivery was 22 days, and the neonatal mortality rate was 39 deaths/1000 live births. In both groups the need for transfusion and the incidence of fetal distress were not increased by the use of tocolytic agents. Among the 171 combined patients who underwent tocolysis, no maternal morbidity related to the tocolytic agents was found and no stillbirths occurred after admission. The neonatal deaths were all related to complications of prematurity. CONCLUSIONS: This is the largest series to date evaluating the use of tocolytic agents in preterm patients with third-trimester bleeding. From these data there does not appear to be any increased morbidity or mortality associated with tocolytic agent use in a controlled tertiary setting. A prospective randomized trial would be necessary to determine whether tocolytic use carries any benefits. PMID- 10368504 TI - The diagnosis and classification of gestational diabetes mellitus: is it time to change our tune? AB - OBJECTIVE: This study was designed to determine the impact on our population of adopting the Carpenter and Coustan criteria for gestational diabetes mellitus in place of the currently used National Diabetes Data Group criteria, to review the evidence supporting replacement of the National Diabetes Data Group criteria with the Carpenter and Coustan criteria, and to propose analogous diagnostic criteria for diabetes in pregnant and nonpregnant women. STUDY DESIGN: The National Diabetes Data Group criteria and the proposed Carpenter and Coustan criteria were both used to retrospectively review medical records of patients screened for gestational diabetes mellitus during 1995 and 1996 in the Kaiser Permanente Northwest Division. Computerized search was performed on automated data systems and software was used for statistical analyses. A MEDLINE review of relevant literature was conducted. RESULTS: Of 8857 pregnant women screened for gestational diabetes in 1995 and 1996, 284 (3.21%) met the National Diabetes Data Group criteria, whereas 438 (4.95%) met the Carpenter and Coustan criteria. We estimate that in our population use of the Carpenter and Coustan criteria in 1996 could at best have reduced the prevalence of infants weighing >/=4000 g from 17.1% to 16.9% and the prevalence of infants weighing >/=4500 g from 2.95% to 2.91%. CONCLUSIONS: Replacing the National Diabetes Data Group criteria with the Carpenter and Coustan criteria would increase by 54% the number of pregnant women with a diagnosis of gestational diabetes mellitus and would also increase costs, while only minimally affecting prevalence of infant macrosomia. The medical literature does not provide compelling evidence for adopting the Carpenter and Coustan criteria. Standardization of both measurement of venous plasma glucose level and diagnostic criteria for gestational diabetes mellitus is an important goal. Parallel criteria for diagnosis and classification of diabetes mellitus in pregnant and nonpregnant women should be developed. PMID- 10368506 TI - Prenatal purified protein derivative skin testing in a teaching clinic with a large Hispanic population. AB - OBJECTIVE: The purpose of this study was to evaluate a tuberculosis skin testing program in an outpatient obstetric teaching clinic. STUDY DESIGN: This was a retrospective chart review and economic analysis. Medical records of all patients in the clinic (n = 1763) who were delivered of infants between 1993 and 1997 were reviewed. Fifty-four percent of the patients were Hispanic, and 38% were white. Epi Info 6.0 (chi2 analysis) was used to calculate statistical significance. RESULTS: Overall, 15.2% of the skin test results were positive. Positive test results were found in 31.3% of Asians, 23.9% of Hispanics, 9.3% of black patients, and 4.1% of white patients. Hispanics had a relative risk for a positive purified protein derivative of 5.9 (3.9 to 8.8) compared with white patients.No chest x-ray films suggested active disease. CONCLUSION: Because the highest incidence of tuberculosis is in people of reproductive age, an effective tuberculosis screening protocol during pregnancy would be valuable in reducing the incidence of tuberculosis nationally. Projected savings of this program were $57,628. PMID- 10368507 TI - Hemodynamic patterns of women with chronic hypertension during pregnancy. AB - OBJECTIVE: The purpose of the study was to evaluate the hemodynamic changes during an initial and subsequent gestation of 15 patients with chronic hypertension. STUDY DESIGN: Data on mean arterial pressure, rapid ejection time, and pulse wave arrival time, recorded at 26 weeks and 37 weeks, were correlated with birth weight and gestational age. Hemodynamic data were acquired with a noninvasive cardiovascular monitor, and blood pressure was recorded with an oscillometric technique. Statistical analyses were performed with the paired Student t test and simple correlation. RESULTS: The birth weights increased in 9 patients and decreased in 6, although more babies were delivered before 37 weeks in the subsequent pregnancy (7 vs 4 patients). No differences were found between the groups of index and repeated pregnancies or among the subgroups whose infant birth weights increased or decreased, except for the pulse wave arrival time at 37 weeks (172 +/- 21 ms vs 143 +/- 29 ms, P <.05). However, strong correlations emerged between the initial pregnancy's rapid ejection time versus the mean arterial pressure at week 37 (r = 0.81, P =.0013) and the repeated pregnancy's rapid ejection time at week 26 versus the gestational age (r = -0.95, P =.00003). CONCLUSION: These observations suggest that rapid ejection time, an empiric indicator of vascular compliance, may have a role in the evaluation and management of hypertension in pregnancy. PMID- 10368508 TI - Terbutaline. PMID- 10368510 TI - All cases of intrauterine fetal death should be evaluated for parvovirus B19 viral deoxyribonucleic acid. PMID- 10368513 TI - Intrauterine scarring as a result of total endometrial ablation could delay the diagnosis of endometrial cancer. PMID- 10368516 TI - Rectal misoprostol in the prevention of postpartum hemorrhage. PMID- 10368517 TI - "Postmenopausal" diagnosis of testicular feminization. PMID- 10368518 TI - Use of oral contraceptives by women who smoke. PMID- 10368519 TI - Benefits and risks of oral contraceptives. AB - The major benefits of modern low-dose oral contraceptives include relative safety and a high degree of efficacy, decreasing the need for abortion or surgical sterilization; reduced risks of bacterial (but not viral) pelvic inflammatory disease and of endometrial and ovarian cancer; improved menstrual regularity, with less dysmenorrhea and blood flow; and, when low-dose combination (not progestogen-only) oral contraceptives are used, reduced acne and hirsutism. Major risks are cardiovascular. Preliminary data from nonrandomized studies suggest that oral contraceptives containing third-generation progestogens are associated with increased risk of venous thromboembolism, particularly in carriers of the coagulation factor V Leiden mutation. The risk of arterial thrombosis, such as myocardial infarction or stroke, may be directly related to estrogen dose, particularly in women who have hypertension, smoke, or are >35 years old. Considering that only users aged >/=30 years who smoke >/=25 cigarettes/d have a higher estimated mortality rate than that of pregnant women, the benefits of oral contraceptives appear to outweigh their risks. PMID- 10368520 TI - Cardiovascular disease: pathogenesis, epidemiology, and risk among users of oral contraceptives who smoke. AB - Smoking increases the risk of lung cancer and cardiovascular disease among persons of both sexes. The risk of cardiovascular disease is further increased among users of oral contraceptives who smoke, particularly those who are >/=35 years old or carry the coagulation factor V Leiden mutation. Other important cardiovascular disease risk factors in women include waist/hip girth ratio >0.8, high concentration of low-density lipoprotein cholesterol (>115 mg/dL), high triglyceride level (>/=150 mg/dL) with low concentration of high-density lipoprotein cholesterol (/=100 mg/dL, hypertension, lack of physical activity, and high-fat diet. Most excess cardiovascular disease among users of oral contraceptives is due to thrombosis (not atherosclerosis); studies indicate that the lower the oral contraceptive estrogen dose is, the lower is this risk. Oral contraceptives containing the third-generation progestins desogestrel and gestodene have been associated with greater risks of venous thromboembolism than are associated with older progestins, although there is some controversy surrounding these findings. PMID- 10368521 TI - Smoking and use of oral contraceptives: impact on thrombotic diseases. AB - OBJECTIVES: The study was intended to evaluate the effects of oral contraceptives and smoking on the risks of arterial and venous thromboembolic diseases among young women. STUDY DESIGN: The study included a survey of data from published epidemiologic studies and evaluation of registry records of all Danish women discharged from the hospital from 1980 through 1993 after a first thromboembolic event. Questionnaires returned by survivors of such events and by control women during the period from 1994 through 1995 were analyzed. RESULTS: In the 1980-1993 data the absolute risk of thrombotic diseases was seen to increase rapidly with age-exponentially for acute myocardial infarction or cerebral thromboembolic attack, linearly for venous thromboembolism-with risks of arterial diseases exceeding those of venous diseases. In the 1994-1995 data the relative risk of thrombotic diseases was seen to increase among users of oral contraceptives irrespective of age. Risk of venous thromboembolism (but not of acute myocardial infarction or cerebral thromboembolic attack) declined as duration of current oral contraceptive use lengthened, risk of acute myocardial infarction or cerebral thromboembolic attack was significantly decreased as ethinyl estradiol doses were reduced, and the relative risk (compared with nonusers of oral contraceptives) for arterial thromboembolic disease among users of desogestrel or gestodene (in conjunction with midrange or low doses of ethinyl estradiol) was lower than the relative risk among users of second-generation progestogens (in conjunction with midrange doses of ethinyl estradiol). The combination of smoking with oral contraceptive use may have a synergistic effect on risks of acute myocardial infarction and cerebral thromboembolic attack (but not of venous thromboembolism), particularly among users of high-dose (50 micrograms) ethinyl estradiol preparations. CONCLUSION: Among the formulations currently marketed in Denmark, where only the progestins desogestrel and gestodene are available with low-dose (20 micrograms) ethinyl estradiol (and only desogestrel was available in that form at the time of our studies), we prefer these third-generation oral contraceptives for smokers. We might also consider such oral contraceptives for women >35 years old as long as they had no other risk factors for thrombotic arterial diseases. PMID- 10368522 TI - Effects of smoking on prostacyclin formation and platelet aggregation in users of oral contraceptives. AB - OBJECTIVE: The aim of this review was to determine which subgroups within the population of smokers and oral contraceptive users are at especially elevated risk for thromboembolic events. STUDY DESIGN: This review covers 10 articles published between 1981 and 1996 that examined the effects of smoking and oral contraceptive use, in conjunction or independently, on factors affecting the coagulation pathway, particularly the expressions of prostacyclin and thromboxane. RESULTS: Heavy, prolonged, or current nicotine use was associated with a reduction in the urinary metabolite of prostacyclin (prostaglandin I2) in oral contraceptive users. Smoking and increased excretion of thromboxane were also linked, and in 1 study the effect was dose related. These changes were associated with increased platelet aggregation. Oral contraceptive use and concurrent smoking increased the risk of acute myocardial infarction by a ratio of 10.1. Although most of this risk was seen among smokers who used second generation oral contraceptives (odds ratio 11.1), with a much reduced odds ratio for smokers who used third-generation oral contraceptives (odds ratio 3.1), the study was not controlled for estrogen dose. A reduction in myocardial infarction risk compared with that in the 1970s was seen for all oral contraceptive users, probably because of the reduced hormonal doses in current preparations. CONCLUSION: Smoking, not oral contraceptive use, constitutes the greater cardiovascular risk. However, cigarette smoking and oral contraceptive use act synergistically to increase the risk of thromboembolic events. Differences in oral contraceptive formulations may mitigate the increased risk resulting from concurrent smoking and use of oral contraceptives, but whether the progestin component or the lowered estrogen dose is responsible is unclear. PMID- 10368523 TI - Hemostatic effects of smoking and oral contraceptive use. AB - This review addresses current knowledge of the effects of lower dose oral contraceptives (containing 35, 30, or 20 micrograms of ethinyl estradiol) on hemostasis in smoking and nonsmoking women. Evidence suggests that formulations containing 30 and 35 micrograms ethinyl estradiol induce a significant activation of coagulation, whereas oral contraceptive preparations with 20 micrograms ethinyl estradiol appear to have a negligible effect or no effect. In nonsmokers who take oral contraceptives any procoagulatory effects that may occur are counterbalanced by fibrinolytic effects. In smokers, however, compensatory fibrinolytic effects to offset the procoagulatory effects seen with 30-micrograms ethinyl estradiol oral contraceptive formulations are absent, shifting the hemostatic profile toward a hypercoagulable state. This suggests that a formulation with the lowest dose of ethinyl estradiol may be most suitable for smokers who wish to use this form of contraception. PMID- 10368524 TI - Effects of oral contraceptives on hemostasis and thrombosis. AB - OBJECTIVE: The object of the study was to determine the effects of oral contraceptives on blood coagulation, in particular on the protein C pathway. STUDY DESIGN: Plasma samples from healthy men, from healthy female users and nonusers of oral contraceptives, and from heterozygous and homozygous male and female carriers of the factor V Leiden mutation (some of whom used oral contraceptives) were tested for their sensitivity to activated protein C by means of a new activated protein C resistance test developed in our laboratory. This assay is based on measurement of the effect of activated protein C on the endogenous thrombin potential, the time integral of thrombin generation initiated in plasma through the extrinsic coagulation pathway. RESULTS: The normalized activated protein C sensitivity ratio ([ETP+APC/ETP-APC]plasma/[ETP+APC/ETP APC]normal plasma, where ETP is endogenous thrombin potential, +APC is with activated protein C, and -APC is without activated protein C) of men was lower than that of healthy female nonusers of oral contraceptives. The normalized activated protein C sensitivity ratio of the users of oral contraceptives was significantly higher than that of nonusers of oral contraceptives. The normalized activated protein C sensitivity ratio of women who were using oral contraceptives with third-generation progestogens was higher than that of users of oral contraceptives with second-generation progestogens. Furthermore, the normalized activated protein C sensitivity ratio of 80% of the users of third-generation preparations fell within the 5th to 95th percentile of the normalized activated protein C sensitivity ratio of female carriers of factor V Leiden, a mutation that is associated with hereditary resistance to activated protein C and with an increased risk of venous thromboembolism. CONCLUSION: Acquired activated protein C resistance may explain the increased risk of venous thromboembolism among users of oral contraceptives reported in epidemiologic studies and the higher risk of venous thromboembolism among users of oral contraceptives with third- versus second-generation progestogens. PMID- 10368525 TI - Oral contraceptives and smoking, current considerations: recommendations of a consensus panel. AB - In a closed meeting, members of the consensus panel evaluated the presentations of the scientific panel and developed a series of recommendations. They outlined clinical imperatives related to the identification and education of patients who smoke, the physician's role in smoking cessation, and the prescription of oral contraceptives for patients who smoke. They also outlined research objectives for the future. The most important suggestions include the following: All patients should be asked about their smoking status at every visit, and all smokers should be encouraged and helped to quit. The decision to prescribe an oral contraceptive requires a detailed personal and family history of thrombotic disease. Measurement of lipid profile should be considered, along with exercise and dietary intervention, for smokers >35 years old who use or request oral contraceptives. Patients >35 years old who smoke heavily (>15 cigarettes/d) should be denied the use of oral contraceptives. Preliminary data suggest that an oral contraceptive with the very low dose of 20 micrograms ethinyl estradiol may be safer for oral contraceptive users who smoke, even for those >35 years old who have an occasional cigarette, but these laboratory findings require clinical corroboration. PMID- 10368526 TI - Prevention of stroke. Advice from experts. PMID- 10368528 TI - Vacuuming away heart attacks. PMID- 10368527 TI - Antibiotics and heart disease. The story heats up. PMID- 10368529 TI - Chronic cough associated with increased heart-attack risk. PMID- 10368530 TI - Ask the doctor. I have tried - and stopped - almost every medication known to man for treatment of high blood pressure. All of them have caused a serious side effect, impotence. Is there any medication out there or coming soon that can control blood pressure without causing impotence? PMID- 10368531 TI - Hepatitis C. Shedding light on the shadow epidemic. PMID- 10368533 TI - Mild cognitive impairment: stumbles on memory lane. PMID- 10368532 TI - Cancer vaccines: of trials and T cells. PMID- 10368534 TI - Arthritis relief: considering the alternatives. PMID- 10368535 TI - By the way, doctor... I've had a bothersome cough for years, and my doctor hasn't been able to make a diagnosis. Two chest x-rays have been normal. I find that coughing can produce embarrassing interruptions when I'm on the phone or in ordinary conversations. My family is worried about what might be going on. I'm not a smoker, so this isn't a smoker's cough I am talking about. Can you help me? PMID- 10368536 TI - Memories lost and found - part I. PMID- 10368537 TI - How useful for psychotherapists are randomized controlled experiments? PMID- 10368538 TI - Men, women, and their symptoms. PMID- 10368539 TI - What are the risks of benzodiazepine dependence? PMID- 10368541 TI - Acupuncture. PMID- 10368540 TI - Malignant melanoma: the dark side of sunshine. PMID- 10368542 TI - Biking and sex: can riding cause impotence? PMID- 10368543 TI - Race and prostate cancer. PMID- 10368544 TI - I am a 54-year-old man with a history of kidney stones. My doctor recently sent me for a "spiral CT scan." What is the difference between a spiral scan and a regular CT scan? PMID- 10368545 TI - Five new ways to look at cancer. PMID- 10368546 TI - Progesterone. PMID- 10368547 TI - Prevention and treatment. Stroke. PMID- 10368548 TI - Nutrition. Vitamin D update. PMID- 10368549 TI - Another weight-loss drug approved. PMID- 10368552 TI - Age-related prevalence of down syndrome. PMID- 10368551 TI - Appropriate model testing. PMID- 10368554 TI - Mifepristone and methotrexate: the combination for medical treatment of ectopic pregnancy. PMID- 10368557 TI - Proceed with caution: understanding and changing norms. PMID- 10368558 TI - College alcohol use: a full or empty glass? AB - Data from the Harvard School of Public Health College Alcohol Study (1993) were used to describe weekly alcohol consumption and its associated problems among a representative national sample of college students. The median number of drinks consumed/week by all students, regardless of drinking status, was 1.5. When students were divided by drinking pattern, the median number of drinks/week was 0.7 for those who did not binge drink and 3.7 for those who did so infrequently. For frequent binge drinkers, the median was considerably higher: 14.5 drinks/week. Nationally, 1 in 5 five college students is a frequent binge drinker. Binge drinkers consumed 68% of all the alcohol that students reported drinking, and they accounted for the majority of alcohol-related problems. The data indicate that behavioral norms for alcohol consumption vary widely among students and across colleges. Therefore, it may not be possible to design an effective "one size fits all" approach to address college alcohol use. PMID- 10368559 TI - Misperceptions of the norms for the frequency of alcohol and other drug use on college campuses. AB - Data from surveys of students representing 100 diverse college campuses were used to investigate the difference between the self-reported frequency of a drug's use and students' perceptions of the frequency of use. Students were asked about the frequency of their own use of 11 drugs (alcohol, tobacco, marijuana, cocaine, amphetamines, sedatives, hallucinogens, opiates, inhalants, designer drugs, and steroids) and how often they thought "the average student" on their campus used these drugs. Respondents typically misperceived their peer norms (designated as the median of self-reported use) by substantially overestimating how often the average student used each drug, both in campus samples where abstinence or infrequent use were the median of self-reports and in samples where the median of self-reports revealed more frequent use. To the extent that they may promote or reinforce students' actual use, these misperceptions should be considered in designing college drug prevention programs. PMID- 10368561 TI - Driver compliance with stop signs at pedestrian crosswalks on a university campus. AB - Pedestrians on college campuses interact continuously with various motorized vehicles. Rates of compliance with stop signs at pedestrian crosswalks and noncomplying vehicles were monitored in spring 1996 on a large urban campus. The number of pedestrians, pedestrian clearance, type of vehicle, hour of day, and day of week were monitored at 3 pedestrian crosswalks. The overall compliance rate for stop signs was 22.8 per 100 vehicles, ranging from 1.4 per 100 for bicycles to 46.2 per 100 for commuter vans. Compliance increased to 53 per 100 vehicles when pedestrians were present in the crosswalk. Several differences in compliance rates were found among the observation sites. Lowest compliance was observed for bicycles and motorcycles. Pedestrians on this and other college campuses risk injuries because of violations of pedestrian right-of-way laws. The problem calls for appreciable educational efforts by college health personnel. PMID- 10368560 TI - Examining the use of tobacco on college campuses. AB - The authors used the Health Risk Behavior Survey for University Students to assess the prevalence of tobacco use among undergraduates in the Florida state university system. They examined the relationships of gender, marital situation, and minority status to 6 smoking behaviors (tried cigarettes, smoked regularly, tried to quit smoking, age when first smoked regularly, number of cigarettes smoked in the last month, and number of days smoked in the past month). Findings suggested that White students were more likely than minority students to try cigarettes and women more likely than men to smoke regularly. Married students smoked more regularly than others and were less likely than single students to have tried to quit smoking. The investigators suggested analyzing latent behaviors associated with smoking and called for a national meta-analysis of data from smoking studies to help clinicians deal with student tobacco use. PMID- 10368562 TI - The effects of an intervention campaign to enhance seat belt use on campus. AB - The national health objectives for the year 2000 called for an increase in the use of safety restraints to 85% of motor vehicle occupants. An assessment on one campus indicated that only 79% of those observed were wearing seat belts. Nursing faculty and students undertook a multimodal intervention campaign to increase seat belt use in the campus community. Observed use of seat belts increased to 81% after the week-long intervention consisting of reminder banners, media coverage, permanent reminder signs, roll-over demonstrations, a presentation on the need for seat belt use, and distribution of seat belt use pledge cards. Although the increase was small, it was statistically significant and could represent considerable savings in healthcare costs if even 2% of the population could be saved from serious injury by using seat belts. In addition, the change in seat belt use represented a decline of nearly 10% in the number of nonusers. PMID- 10368563 TI - Combating sexual offenses on the college campus: keys to success. AB - The authors emphasize the need for programs to prevent sexual offenses at institutions of higher education. They briefly describe efforts underway at Bowling Green State University and offer 6 strategies for improving the likelihood of success in sexual assault education, prevention, and response efforts. PMID- 10368564 TI - La maladie du petit papier. PMID- 10368565 TI - [Nutritional status, obesity and metabolic control in children with type 1 diabetes mellitus]. AB - BACKGROUND AND AIMS: The aim of this study was to evaluate the nutritional status of children with type 1 diabetes and to search for possible influences of changes in body composition on aspects of diabetes. METHODS: A group of 96 diabetic subjects (41 males and 55 females) were studied, aged between 3 and 19 years old. The following parameters were examined: weight, stature, 5 skin folds, 7 circumferences, bioelectric impedance, arterial pressure, cholesterolemia, triglyceridemia, insulin dose, HbA1c and duration of disease. RESULTS: Obesity and overweight were present in 34.5% of the sample, but obesity was only observed in females (25.5%). There was also a high percentage of underweight subjects (11.5% of the entire sample). The mean values of weight BMI, 5 skin folds, 4 circumferences, FM (calculated using fold measurement and BIA) and AFA were higher in females, whereas mean values of waist/hip ratio and waist/thigh ratio and FFM (in % of body weight) were higher in males. A close correlation was also found between the 4 weight classes (underweight, normal weight, overweight, obese) and the majority of marker parameters for adiposity (5 folds, 4 circumferences, BIA, FM calculated using BIA, fold measurement and AFA). Of the other parameters examined (mean duration of disease, HbA1c assay, daily insulin dose, total cholesterolemia, triglycerididemia, arterial pressure), only the daily insulin dose showed higher values in females in 3 weight classes (underweight, normal weight and obese). Following a comparison with the control population (2469 subjects), higher mean values were found in the latter compared to diabetic subjects, but only in relation to 3 skin folds (tricipital, subscapular and suprailiac) and one circumference (forearm). CONCLUSIONS: The study shows a high frequency of overweight and obesity in children with type 1 diabetes, comparable to that in the healthy population. The finding of a higher frequency of obesity in diabetic females might be explained by their advanced puberal status, given that almost all the obese diabetic females were aged between 10 and 19 years old. The study confirms the validity of a number of anthropometric measurements (BMI, folds, circumference) and BIA in the evaluation of nutritional status in terms of body composition. PMID- 10368566 TI - [Anti-pseudomonas-specific precipitins as a marker of chronic infection in patients with cystic fibrosis]. AB - AIMS: The authors underline the characteristics of Pseudomonas aeruginosa and its methods of action on bronchial mucosa in cystic fibrosis. They then discuss the two concepts of "colonisation" and "chronic infection". METHODS: The level of "infection" was evaluated using an immunoelectrophoretic method involving the precipitation of bands of specific precipitins. A technical description of the method is included. The authors illustrate the use of the method in 78 cases of cystic fibrosis, comparing the positive results obtained using precipitin electrophoresis with the results of direct bacteriological findings. RESULTS: Using the bacteriological criteria, a total of 26.9% of patients were diagnosed as infected, whereas this percentage rose to 32.1% using precipitins. 87.2% of cases were concordant using both methods. CONCLUSIONS: As a practical solution, the authors recommend that the two methods are combined, thus obtaining a marked reduction in the number of false positives with obvious consequences in terms of therapeutic decisions. PMID- 10368567 TI - [Urinary calcium excretion in a population of children living in Southern Italy]. AB - BACKGROUND: Very few studies have evaluated the role of urinary calcium excretion as marker of bone metabolism in children. Normative data are lacking in these age groups. METHODS: In a group of 122 children (66 females e 56 males), mean age 108.13 +/- 18.73 months, attending a primary school in Nocera Superiore (Southern Italy) the following parameters were evaluated: sex, age, weight, height, BMI (weight/height2), urinary calcium excretion which was measured as the urinary calcium/creatinine concentration ratio (Uca/cr) in extemporaneous sample of the second urine in the morning. The Uca/cr ratio was not normally distributed in this childhood population. Natural logarithmic transformation was used to analyze the data. RESULTS: The mean value of Uca/cr was 0.125 +/- 0.102 mg/mg. A significant correlation was found between the Uca/cr ratio and sex. The Uca/cr ratio values were higher in girls than in boys. Mean Uca/cr was 0.144 +/- 0.113 mg/mg in females while it was 0.102 +/- 0.083 mg/mg in males ("t"-test: p < 0.05). No correlation was found between the Uca/cr ratio and other variables such as age, height, weight and BMI either in the simple or in the multiple linear regression analysis correcting both for the sex and the age. Data by age-group and sex were reported as percentile tables. CONCLUSIONS: The discussed variations of the Uca/cr in children from different areas and sex confirm the need for reference standards calculated in the different population groups. PMID- 10368568 TI - Practical applications of monitoring respiratory mechanics in newborn. AB - The authors evaluate the use of data supplied by common systems used to calculate respiratory mechanics in newborns in order to analyse the impact on the management of patients requiring assisted ventilation. Over the past few years, the sale of respiratory monitors that are easier to use and less complicated to manage, as well as being less expensive, has meant that nearly every Neonatal ICU can now determine, in real time, both the causes of respiratory insufficiency and the conditions of the newborn lung during mechanical ventilation to ensure improved adaptation of ventilatory support. Leaving aside a discussion of respiratory physiopathology and the principles regulating pulmonary ventilation, the authors focus on the practical effect that data normally supplied by this type of apparatus (airway pressure, inspiration and expiration airflow, expiration minute volume calculated for each respiratory act, tidal volume, system leaks and dynamic compliance) have on the ventilatory "setting" chosen by the clinician or on the diagnosis of some commonly found situations during neonatal mechanical ventilation. The experience described concerns the impression of clinicians, skilled in the management of patients receiving conventional artificial ventilation but not so attracted by respiratory physiopathology, regarding the value of such instruments. The unanimous positive opinion confirms the thesis whereby respiratory monitoring can certainly help the neonatologist make a real time evaluation of both the causes leading to respiratory failure and the effect on the lung of any therapeutic decisions. PMID- 10368569 TI - The Smith-Magenis syndrome: a new case with infant spasms. AB - The Smith-Magenis syndrome (SMS) is characterized by congenital anomalies, mental retardation and the interstitial deletion of the 17p. 11.2 chromosome. The subjects affected by this syndrome show cranio-facial dysmorphias, brachycephalia, skeletal, ocular, cardiac, genitourinary and otolaryngological anomalies. The central nervous system is affected and this may be shown by psychomotor retardation, intellective deficit, electroencephalographic alterations (reduced/missing REM phase); the neuroradiological tests detect megacisterna magna, cerebellar hypoplasia, cortical dysplasia, ventricular asymmetry. Behavioural troubles are frequent and, among them, self-aggressive conducts (tearing out the nails). The syndrome is associated with the interstitial deletion of the 17p. 11.2 chromosome. The diagnosis can be made in the pre-natal period and a mosaic situation is possible. Even though the cases of SMS reported in the literature allow defining a characteristic phenotype, studies have been carried out to quantify the deletion of the chromosome 17 in order to identify the chromosomic tract which is responsible for the phenotypical induction. The deletion can either appear de novo or come from one of the parents. In addition, these subjects can show peripheral neuropathy, missing or reduced deep tendon reflexes and (rarely) epileptic crises. However, by reviewing the literature, no descriptions of patients affected by infant spasms are pointed out. This report refers to a new case of Smith-Magenis syndrome in a nine-month old girl with spasms in extension. PMID- 10368571 TI - Induction and pharmacological treatment of oral osteopenia in rats. AB - BACKGROUND: Osteoporosis is a major public health problem, responsible for a great number of fractures, associated with devastating costs to society. In addition, oral bone loss has an enormous impact on the health quality of life of patients, affecting up to 90% of elderly individuals. The aim of this work was to elaborate an animal model of mandibular and maxillary osteoporosis in which to evaluate bone loss and possible prevention by pharmacological treatment. METHODS: Six Sprague-Dawley rats were gonadectomized and treated with clodronate (male) or 17 beta-estradiol (female) for two months. Six gonadectomized and six sham operated rats of both sexes were treated with placebo. The mandible and maxilla, fixed and methacrylate embedded, were serially sectioned, microradiographed and processed for histomorphometric analysis. RESULTS: Gonadectomy did not modify the amount of compact and trabecular bone in mandibles of rats of either sex, treated or not with clodronate or estrogens, compared to sham-operated rats. Compared to sham-operated rats, a 10-25% increase of bone porosity was found in the maxilla of ovariectomized rats, either receiving estrogens or not, while in male rats no difference among groups could be evidenced. CONCLUSIONS: The conclusion is drawn that rats, due to their peculiar masticatory habits yielding huge loads on oral bones, do not represent a suitable experimental model for studying oral bone loss related to skeletal osteoporosis. In order to worsen oral osteopenia it would be mandatory to combine gonadectomy with a mechanical unloading (i.e. after molar extraction) of mandibular or maxillary bone. PMID- 10368570 TI - [Science, customs and social life in the Middle Ages]. PMID- 10368573 TI - Anatomical study of the temporal fasciae and fat pads. AB - BACKGROUND AND AIM: Although a great number of studies have been published on the anatomy of the various fascial layers in the temporal region and the interposed fat pads, there is still uncertainty regarding the organization of the temporal fasciae, above all at the level of the zygomatic arch, and the relationships between the latter and the frontal branch of the facial nerve. This study aimed to describe the anatomy of the temporal fasciae and their relationships both with the interposed fat pads and with the frontal branch of the facial nerve. METHODS: The study was carried out in 10 heads from fresh cadavers which were used to dissect the different tissue layers at the level of the temporo-zygomatic arch. RESULTS: The results of the dissections made in this study confirm the existence of three over-lying fascial layers (superficial temporal fascia, intermediate temporal fascia and deep temporal fascia), interposed by the same number of fat pads (superficial, intermediate and deep). The frontal branch runs below the superficial fat pad in close contact with the periosteum of the zygomatic arch. CONCLUSIONS: The authors suggest adopting a single nomenclature to describe the fasciae and the fat pads in the temporal region. PMID- 10368574 TI - [Diagnostic criteria of Sjogren syndrome]. AB - The present knowledge about Sjogren classification, diagnosis, therapy and its prognosis is reviewed. Some researchers about immunological and histological factors are also underlined. Finally, a complete picture of this syndrome is given. PMID- 10368572 TI - [Radial forearm free cutaneous flap. Evaluation of the level of damage to the donor site]. AB - BACKGROUND: After more than a decade of experiences, the radial forearm fasciocutaneous free flap has proven to be a well standardized surgical technique, widely used for its easy application and versatility. The review of the literature shows that most of the contraindications to the use of this flap concern mainly the donor site morbidity. Aim of this work is to evaluate the complications of this surgical technique, to study the functional impairment and the scars secondary to the harvesting of the flap, in order to point out which techniques can contribute to their reduction. METHODS: Seventeen patients submitted consecutively to oral cavity reconstruction using a radial forearm flap, whose donor site has been reconstructed with a full-thickness skin graft have been studied. The postoperative complications have been examined, the functional and aesthetic alteration at the donor site analyzed, with a medium follow-up of 17.5 months. RESULTS: The study of this series has pointed out not relevant early and middle term postoperative complications, no significant hand or finger motility and sensorial deficiencies associated with a percentage of full aesthetic and functional satisfaction of patients for this treatment higher than 80%. CONCLUSIONS: On the basis of personal clinical experience, even if on a small series of cases, the conclusion is drawn that the functional and aesthetic outcomes depend significantly on the surgical techniques used to harvest the flap and to cover the donor site. PMID- 10368575 TI - [Dentinogenesis imperfecta. Scanning electron microscopic study and microanalysis]. AB - BACKGROUND: Dentinogenesis imperfecta (DI) is an inherited dentine defect which affects both the primary and secondary dentitions. Shields et al. in 1973 suggested a classification of DI within three types: type I, associated with osteogenesis imperfecta (OI), type II and type III. Although the varying clinical, radiographic and histological findings in DI have been described in detail, an available method for closer examination of the abnormal dentine matrix, electron microscopy, has seldom been used. Scanning and transmission electron microscopy studies can help to understand the pathogenesis of the different types of heritable dentine defects and the diagnosis and classification of these diseases. The aim of the present study was to evaluate a case of DI using scanning electron microscopy and microanalysis. METHODS: Dentine was obtained from tooth samples from a fourteen-year-old boy affected by DI and from third molars extracted for therapeutic reasons used as controls. Samples were observed with a scanning electron microscope, scanning electron micrographs were evaluated with an image analysis program and specimens were finally observed with a scanning electron microscope equipped for micro-analysis. RESULTS AND CONCLUSIONS: The results obtained showed that the total number of dentinal tubules was significantly reduced and the presence of a dentine mineralization defect in the patient affected by DI, in comparison to the controls. PMID- 10368576 TI - [Maxillo-mandibular fixation by mono-cortical screws. Clinical indications and surgical methods]. AB - BACKGROUND: Intermaxillary fixation is one of the most reknown and widely used techniques in maxillofacial traumatology. It's carried out usually by means of direct criss-cross teeth wiring or through the wiring of a metallic archbar on the upper and lower jaws. These techniques are time-consuming operations, they can produce dental or periodontal damages, and are not well tolerated by the patient, even under local anesthesia. Recent experiences in oral implantology and in the use of miniscrews for rigid internal fixation suggest the experimentation of new, easy to use and better tolerated systems for bone-anchored intermaxillary fixation. METHODS: 1-0 stainless steel wires and titanium monocortical screws, 2 mm of diameter and 12 and 15 mm of length, have been used as alveolar-bone anchorages for the intermaxillary fixation of 10 mandibular fractures. The fixations have been performed either under general or local anesthesia, with 2, 4 or 6 points of alveolar bone anchorage, maintaining the fixation for 15 days in condylar fractures and for 40 days in all the other cases. RESULTS: A really good compliance of the patient towards all the procedures performed under local anesthesia, with a clear reduction of postoperative discomfort has been observed. Infection or rejection of the implanted screws did not occur as well as cases of alveolar or dentoparodontal damages. CONCLUSIONS: This preliminary report on a new intermaxillary fixation technique didn't point out any significant complication of the procedure, showing at the same time that this technique can be easily performed under local anesthesia on out-patients with a better compliance, lower postoperative discomfort and good skeletal stability. PMID- 10368577 TI - [The value of leukotriene receptor antagonists in the therapy of bronchial asthma one year following the introduction of Montelukast. Symposium at Munich, 6 March 1999]. PMID- 10368578 TI - [International cooperation in world cardiology: the role of the World Heart Federation]. PMID- 10368579 TI - [The appearance of giant negative T waves in anterior acute myocardial infarct with a Q wave is associated with minor myocardial damage and a minor extension of coronary disease]. AB - INTRODUCTION AND OBJECTIVE: The early inversion of T waves in patients with acute myocardial infarction has recently been related to a better left ventricular function and a more favourable evolution, contrary to what happens in the unstable angina. On the other hand, the significance of the appearance of deep negative T waves in the early phase of some acute myocardial infarction is not known. The aim of this study is to evaluate its relation with the existing myocardial damage and the underlying coronary artery disease extension in anterior some with Q wave. METHODS: 48 patients with a first anterior Q-wave acute myocardial infarction, thrombolized or not, admitted to hospital with an evolution of less than 24 hours, and with a coronariography performed before discharge were analyzed. Giant negative T waves were defined as those which were 8 mm or more from baseline. RESULTS: 17 of the 48 patients presented giant negative T waves (T-group) and 31 did not (N-group). In the T-group patients, the size of the negative T wave was 11.29 +/- 2.86 mm and the number of precordial leads with negative T waves was 4.35 +/- 1.57. There were no differences between both groups in variables such as sex, coronary risk factors, and other basal characteristics. The T-group patients were younger, had lower peak-CK, CK-MB and LDH levels and presented greater recovery of R waves during the follow-up, the differences being significant with the N-group patients. The left ventricular ejection fraction was higher (56.3 +/- 13.4 vs 42 +/- 12%; p < 0.001) and the number of affected coronary vessels was lower in the T-group (1.12 vs 1.64; p < 0.01); there were no differences in the localization or severity of coronary lesions, nor in the frequency of postinfarction myocardial angina. None of the patients in the T-group were Killip > I, while this situation occurred in 38.7% of the N-group patients. CONCLUSIONS: The appearance of giant negative T waves in the acute or early phase of Q-wave anterior acute myocardial infarction is associated with a smaller infarct size, lower functional deterioration and less extension of the underlying coronary disease. PMID- 10368581 TI - [Is prior balloon dilatation necessary before implanting a stent?]. PMID- 10368580 TI - [A direct stent without predilatation. Our experience in 300 lesions]. AB - INTRODUCTION: The stent alone technique, direct stenting without predilatation, aims to reduce cost and procedural time. Other potential benefits are the avoidance of abrupt vessel closure after balloon angioplasty and lessening of the restenosis rate due to the reduced arterial injury. We present our experience with this therapeutic approach in a long series of patients. PATIENTS AND METHODS: 230 patients referred to our unit were included with 300 non-occlusive stenotic lesions without excessive tortuosity, calcification, length or angulation and with a reference vessel diameter > or = 2.5 mm. In these patients stent implantation without predilatation was attempted. The immediate angiographic results and procedural related complications were evaluated. RESULTS: The stent alone technique succeeded in 256 (85%) among the 300 lesions treated. In 43 (14.3%) lesions predilatation was required and in one case the stent could not be positioned. A new dilatation after deployment was required due to suboptimal stent expansion in 27/256 (10.5%) lesions. Stent embolization occurred in 5 patients, 4 stents were retrieved and there were no clinical sequelae. The best results were obtained in non-subtotal and non-bifurcated lesions type A or B1 without moderate calcification, tortuosity or angulation. CONCLUSIONS: Direct stenting is feasible in a large number of patients with a high success rate after an appropriate selection. The most optimal lesions to be treated with this technique are < or = 90% stenotic non-bifurcated lesions type A or B1 without moderate calcification, tortuosity or angulation. PMID- 10368583 TI - [The 4 Provinces Study: its principal objectives and design. The Researchers of the 4 Provinces Study]. AB - INTRODUCTION AND OBJECTIVES: Spain shows an important variation in the geographical distribution of ischaemic heart disease and cerebrovascular disease mortality. In this article, the primary objectives and design of the Four Provinces study are described. In this study we analyzed the contribution of environmental factors (diet, lipid profile and plasma antioxidants), acting in childhood, to explain differences in cardiovascular mortality rates between different provinces in Spain. METHODS: An ecological design was projected in which the units to study were four Spanish provinces with a wide variation in cardiovascular mortality in adulthood. The design compares diet, anthropometric variables and biological markers (particularly plasma lipids and antioxidant levels) in 6-7-year-old children, between the two provinces with the highest cardiovascular mortality rates and the two provinces with the lowest. The information for each province is collected in a cross-sectional manner in a representative sample of children from each province. DISCUSSION: Evidence from the literature concerning Northern European countries suggest the contribution of environmental factors during early age in the development of cardiovascular disease in adulthood. The "Four Provinces" study will provide, for the first time, information about the influence of factors in early childhood of cardiovascular risk in a Mediterranean country. The study will also offer interesting data about food intake during school age in four provinces and it will allow us to estimate values of population of the variables of interest in those provinces. PMID- 10368582 TI - [The radiofrequency catheter ablation of intranodal reentry tachycardia in children and adolescents]. AB - INTRODUCTION: Radiofrequency catheter ablation of slow pathway is the primary nonpharmacological treatment for the atrioventricular node reentrant tachycardia at present. OBJECTIVES: To evaluate the results and long term follow-up of the catheter and radiofrequency modification of the AV node in the treatment of the atrioventricular node reentrant tachycardia in children and adolescents in our center. METHODS AND RESULTS: In a series of fifteen patients, 7 men and 8 women, with a mean age of 8.7 +/- 5.5 years (range, from 4 to 18) with atrioventricular node reentrant tachycardia underwent radiofrequency catheter ablation. Six patients had been treated previously with 1.4 +/- 1.1 antiarrhythmic drugs and nine had not received treatment. In all patients slow-pathway atrioventricular node ablation guided by an anatomic stepwise approach was attempted. In 14 out of 15 patients slow pathway was successfully ablated; and in one patient with a previously failed slow-pathway ablation, a fast-pathway ablation was performed. Tachycardia recurred in one patient, and slow pathway was ablated in a second procedure. After successful slow pathway ablation in 14 patients, the shortest cycle length in which the AV conduction was maintained at 1:1, was increased from 271.3 +/- 22.6 to 316.7 +/- 30.1 ms (p < 0.001), while the AH and HV intervals and shortest cycle length of 1:1 VA conduction remained unchanged. In the patient who had fast pathway ablation the AH interval was increased from 65 to 130 ms, and retrograde VA conduction was lost. Noninducibility of the tachycardia was achieved in all patients without significant complications. During a mean follow up of 18.8 +/- 11.4 months (median of 16), all patients are symptom-free without medication. CONCLUSIONS: Radiofrequency catheter ablation is a successful and safe therapeutic alternative in the treatment of atrioventricular node reentrant tachycardia in children and adolescents. PMID- 10368584 TI - [The activation patterns during atrial fibrillation in an experimental model]. AB - INTRODUCTION AND OBJECTIVES: In atrial fibrillation, along with the mechanisms of complete reentry and random activation focal activation patterns have been described which have been attributed both to propagation from the endocardium and to the existence of zones with automatic activity. The objectives of present study are to analyze and quantify the atrial activation patterns in an experimental model of atrial fibrillation. MATERIAL AND METHODS: In 11 Langendorff-perfused rabbit hearts atrial fibrillation was induced by atrial burst pacing after right atrial dilatation with an intra-atrial balloon. A multiple electrode consisting of 121 electrodes and positioned in the right atrial free wall was used to construct the activation maps corresponding to 10 segments of 100 ms in 11 different episodes of sustained atrial fibrillation (one per experiment). RESULTS: Of the 110 segments analyzed, 44 (40%) corresponded to random activation patterns. Fifteen segments (14%) corresponded to complete reentry, and in these cases the number of consecutive rotations ranged from 1 to 2.25 (mean 1.4 +/- 0.4). In 49 segments (44%) a single activation front was seen to pass through the recording area without block; alternatively, two simultaneous fronts were recorded that did not re-excite the zone activated by the other. In two segments (2%) there was a focal activation pattern without evidence of propagation from the epicardium surrounding the activated zone. CONCLUSIONS: a) in the experimental atrial fibrillation model used, random activation patterns are more frequent than complete reentry patterns; b) complete reentry can occur in areas smaller than 1 cm2, and c) focal activation during atrial fibrillation is rare. PMID- 10368585 TI - [The induction of a septal infarct as a therapeutic alternative in hypertrophic obstructive cardiomyopathy: new observations apropos a case]. AB - The hypertrophic cardiomyopathy may be associated with a variable degree of left ventricular outflow tract obstruction. There are several available therapeutic strategies for treatment: pharmacological, dual-chamber pacing, surgery and induced septal infarction. This last one is a novel technique with less experience in practice. We present the clinical case of a patient which showed persistent and severe obstruction in spite of the medications and dual-chamber pacing, and who underwent this novel technique. The results were optimal but new observations arise from this particular case. PMID- 10368586 TI - [The therapeutic focus in severe hypertrophic obstructive cardiomyopathy with multivessel coronary disease]. AB - The association of severe hypertrophic obstructive cardiomyopathy and coronary artery disease increases surgical morbimortality, even more in patients over 65 years. We describe a combined therapeutic approach to these diseases. A 68-year old woman with a diagnosis of hypertrophic obstructive cardiomyopathy was in functional class IV for angina and dyspnea despite 360 mg of propranolol a day. An echocardiogram and a complete cardiac catheterization were performed under betablocker therapy, confirming a severe hypertrophic obstructive cardiomyopathy and revealing severe stenosis in the proximal left circumflex and the proximal right coronary arteries, and a moderate lesion in the mid-left anterior descendent. They were both treated with balloon PTCA, and a 3 x 15 mm stent was placed in the circumflex and a 3.5 x 20 mm stent in the right coronary, with an excellent angiographic result. A basal hemodynamic study was then performed and A V sequential pacing was attempted, achieving a significant decrease in the left ventricle outflow tract gradient. A DDD-R pacemaker was implanted. Echocardiographic study was performed post-implantation, and follow-up was made six months later with a new coronary angiography, hemodynamic study and a Doppler echocardiogram. At the present time A-V sequential pacing as a therapeutic option for hypertrophic obstructive cardiomyopathy and coronary angioplasty and stenting for the treatment of coronary artery disease are sufficiently established and supported to be offered as a combined therapy to patients suffering from both diseases, specially those with a higher surgical risk. PMID- 10368587 TI - [Pseudoaneurysm of the mitral-aortic fibrosa secondary to the partial detachment of a mechanical aortic prosthesis]. AB - We report a patient with an iatrogenic pseudoaneurysm of the mitral-aortic intervalvular fibrosa under the level of a prosthetic aortic valve. Partial detachment of the proximal suture line of the sewing ring is the most probable etiology. This unusual pathology was well demonstrated by transesophageal echocardiography incidentally in an asymptomatic patient who died of a brain haematoma. Aspects related to the diagnosis, treatment and prognosis of this infrequent finding are discussed. PMID- 10368590 TI - [The publication in biomedical journals of clinical trial protocols: a new way to improve research quality]. PMID- 10368588 TI - [Subpulmonary obstruction caused by an aneurysm of the membranous ventricular septum. A study of 2 cases]. AB - Aneurysms of the membranous ventricular septum can exceptionally produce subpulmonary obstruction. We report two patients, an 8-year-old and a 2-year-old respectively, with a perimembranous ventricular septal defect and subpulmonic stenosis caused by an aneurysm of the membranous septum. Diagnosis was made by Doppler two-dimensional echocardiography and was confirmed by cardiac catheterization and was surgery. It is shown that this obstruction may appear late due to the growth of the aneurysm. In conclusion, we believe that the appearance of an aneurysm on a ventricular septal defect is not always beneficial, as it may result in right ventricular outflow tract obstruction or other kinds of complications which may require cardiac surgery. PMID- 10368589 TI - [Cardiac cysts. A case of isolated cardiac hydatidosis]. AB - In the presence of cardiac cysts we must discard a hydatid disease, even if there is no involvement of other organs. Imaging techniques are useful for guiding the initial diagnosis. The presence of daughter vesicles or multiple cysts is very characteristic. We present a patient affected by cardiac hydatid disease, in the form of multiple cardiac cysts, without extracardiac affectation, who presented pericardial chest pain. The patient was dealt with surgery to avoid the risks of a cyst rupture. PMID- 10368591 TI - [Informed consent in cardiac rehabilitation]. PMID- 10368592 TI - [Targeted enamel erosion/abrasion. The treatment of enamel dysplasia by a microabrasion technic]. PMID- 10368593 TI - [Minimally invasive unreinforced adhesive composite bridges: the clinical procedure]. PMID- 10368596 TI - [The development of third molars in the children of Croatia]. AB - The objective of this research was to investigate the prevalence of third molars in children in Istria aged 7 to 18. There are 2350 panoramic radiographs of children in Istria that have been analysed and then divided into 7 age groups. In each of these groups there was the same number of boys and girls. It can be expected that over 50% of examinees have third morals at ages between 10-11. The development of third morals starts earlier in the mandible, and it is statistically significant that there are more third morals in the mandible before the age of 12 than in the maxilla (p < 0.01). Furthermore, it is statistically significant that nine year old girls have more third morals than boys, but twelve year old boys more than girls. The differences in the prevalence of third morals between the jaw sides were statistically significant only for mandibles of twelve year old girls. Hypodontia of third morals has been found in 4.5% of boys and 5.8% of girls aged 14 to 18. PMID- 10368594 TI - [The determination of the coagulation threshold of the electrolyte potassium chloride in human mixed saliva]. AB - For quantitative assessment of the stability of dispersed phase corpuscles in the gingival fluid, coagulation threshold and coagulating effect of electrolyte KCl were studied in salivary pools of caries-resistant (CR) and caries-liable (CL) subjects by the optic method widely used for investigation of pure colloid systems. Test samples were prepared by mixing certain volumes of saliva, distilled water, and KCl. Light absorbance was measured by photoelectrocolorimetry on a MKMF-1 device and pH was measured by the potentiometric method using pH-121 device. Coagulation threshold and coagulating effect of the electrolyte were estimated using the graphic method. The saliva of both CR and CL subjects generally follows the regularities typical of pure colloid systems. KCl coagulation threshold of CR subjects' saliva is higher than of subjects with caries, while its coagulating effect is higher in the CL group. Dispersed phase corpuscles in the saliva of CR subjects are more stable than in subjects with caries. These results may be used in development of quantitative methods for assessing the stability of salivary dispersed phase corpuscles and salivary mineralizing capacity in subjects with different oral status. PMID- 10368597 TI - [A comprehensive study of the mechanical effects of the long-term and regular use of chewing gum]. AB - The results of regular gum chewing (30 min 3 times a day after meals for 4 weeks) were assessed in 46 volunteers by the functional diagnosis methods. Chronometry of food chewing and arbitrary gum chewing for 30 min was carried out before the investigation. Daily duration of food chewing is about 33 min. During gum chewing, the number of masticatory strikes on the inert side is higher during the first 5 min and then levels. This leads to levelling of mechanical loading of the lateral parts of the jaws and of bioelectrical activity in the masticatory muscles proper. The gum is bit 3-5 times more often with frontal teeth than with lateral teeth. Regular use of chewing gum notably increases the blood supply to the periodontium, and the regulatory mechanisms fail to cope with it in periodontal diseases; this leads to venous congestion in the regional vascular system. PMID- 10368595 TI - [The cytological indices and electrokinetic mobility of the cell nuclei of the buccal epithelium in assessing periodontal status]. AB - Study of electrokinetic mobility of buccal epitheliocyte nuclei, nonspecific resistance of buccal mucosa cells, and cytograms of patients with periodontitis of different severity revealed a clear-cut correlation between these parameters and severity of clinical picture of periodontal inflammation. Morphological and cytological studies of buccal epithelial cells are proposed to be used for assessing the severity of periodontal inflammation and the efficacy of therapy. PMID- 10368598 TI - [The functional activity of the minor salivary glands of the lips in allergic cheilitis]. AB - Functional activity of minor salivary glands is studied in 289 children, 52 of these without signs of cheilitis and 237 with allergic cheilitis at different stages. Three variants of secretion are distinguished: fast, medium, and slow. The ratio of these variants changed in exacerbation of disease in comparison with that in health or remission. The activity of the glands depends on clinical manifestations of cheilitis. Changes in the activity of minor salivary glands determine the clinical picture, localization, and type of involvement in cheilitis. PMID- 10368599 TI - [The choice of fungicidal preparations for the treatment of candidiasis of the oral mucosa and lips]. AB - Therapeutic efficacies of various antifungal agents are assessed in 461 patients. A new method for assessing the sensitivity of Candida to various fungicidal drugs is described, permitting a rapid proper choice of the drug for treating oral candidiasis. PMID- 10368600 TI - [Septic metastatic complications in facial furuncles and carbuncles]. AB - According the information of Maxillo-Facial Surgery Clinic of Moscow Medical Academy the percentage of the patients with face furuncles and carbuncles from all the hospitalised is the following: 1994--4.5%, 1995--7.9%, 1996--11.4%, 1997- 17.0%. That gives about 3.7 times during the last 2 years. The rate of dangerous for life septic metastatic complications in the course of such diseases considerably exceeds those at phlegmons. This figure has increased 2 to 2.5 times during the last 2 years. Two clinical cases which demonstrate possibility of bacterial thromboemboli spreading in face furuncles and carbuncles not only through small, but also through large blood circulation with formation of septic centres in organs are described. The treatment tactics for such patients requires a strictly individual approach. At slightest suspicion of the complication urgent hospitalisation of the patients for active conservative therapy is necessary. In case of purulent process extension surgery is indicated. PMID- 10368601 TI - [The treatment of patients with complicated mandibular fractures using a method for the intraosseous administration of biologically active drug agents]. PMID- 10368602 TI - [The use of the Periotest method for the clinical assessment of the efficacy of dental implantation]. AB - The paper describes a modern diagnostic method: Periotest for assessing the stability of intraosseous implants. The mobility of implants after remote and immediate implantation is assessed in 65 patients with dentition defects. The study yielded complete information on the status of tissue complex, detected the early signs of impaired bone integration after implantation, and helped more reliably predict the results. PMID- 10368603 TI - [The evaluation of the efficacy of using different types of implants]. AB - Published reports on the clinical efficacy and remote results of treatment by dental implants of different types are reviewed. The authors assessed the incidence and causes of complications of implantations on the basis of a long (up to 12 years) follow-up of 865 patients to whom 1155 grafts were implanted. Methods for treating and preventing complications are developed. Implants have to be removed early (up to 1 year postoperation) because of errors in defining the indications to surgery and in the operative technique; in later periods the causes of implant removal are implant and denture fractures and bone resorption round the implant because of functional over-exercise. PMID- 10368604 TI - [Bone tissue density study of the mandible in patients after orthodontic treatment using implants]. AB - Mandibular bone compactness has been studied by ultrasonic osteometry before and after implantation. Ultrasonic examinations were carried out in remote periods after orthodontic treatment (implantations) of patients with dentition defects. Analysis of the results indicates that 2-4 years after the implantation is the critical term, because masticatory loading of dentition is levelled and the unilateral chewing reflex is restructured after fitting with dentures. PMID- 10368605 TI - [A nonpigmented melanoblastoma of the mandibular alveolar process]. AB - A 64-year-old male patient consulted a doctor for a tumor-like formation in the left submandibular area. The dentist detected ulceration of the alveolar process from the V to the VII tooth. Histological examination diagnosed pigment-free melanoblastoma, a tumor extremely rare in the oral cavity; the prognosis in such a tumor is much worse than in skin melanomas. This case confirms a viewpoint that tumors in the oral cavity are difficult to diagnose and they are usually diagnosed after they manifest themselves by metastases. PMID- 10368608 TI - [The restoration of premolars and molars with titanium pin inserts with composite coatings]. PMID- 10368607 TI - [The elimination of the retractility of the skin portion of the nasal septum]. AB - The authors propose a method which decreases the traumatism of surgery for nasal septum repair and helps attain more stable cosmetic results. An open endonasal incision is made behind the alar cartilage pedicles and continued along the fold between the alar and triangular cartilages to the nostril wing base. Perpendicular to this incision and starting from its end, another incision is made, also through the mucosal and cartilagenous tissue to the site where lateral parts of alar cartilages are changed for median. The mucocartilaginous flap is thus cut from the external upper part of the lateral compartment of the alar cartilage. The same intervention is made from the other side. Then the flaps and wound edges are mobilized, the columella is rendered a proper position, and the detected hidden tissue defect is filled with mucocartilaginous flaps. This method is based on correct understanding of the deformation, detection of the latent defect of tissues, and its repair by means of the adjacent tissues making incisions from the nasal cavity. The modification differs favorably from the prototype method. PMID- 10368606 TI - [The use of new biologically compatible materials in filling defects of the jaw]. AB - A method for plastic repair of defects and deformations of the mandible by implants made of glass-crystal material based on magnesium orthophosphate and fluoroapatite is described. Good results and experience gained by the present time in the use of biocompatible crystal glass materials recommends them for wider use. PMID- 10368609 TI - [Laser technologies for the making of dentures, orthodontic appliances, metallic and sapphire implants and root pins]. AB - The authors analyze 10-year experience gained in the use of patented laser technologies for making metal dentures, orthodontic devices, metal and sapphire implants, radical pins, and other structures from various metal dental alloys (steel IXI8H9T, cobalt-chromium, silver-palladium, gold, titanium, etc.). Kvant 155, a new-generation Russian laser device, is described. PMID- 10368610 TI - [A corrosion resistance study of titanium samples obtained by the electron-beam method]. PMID- 10368611 TI - [The probable changes in the structure of the network of state and private dental clinics]. PMID- 10368612 TI - [The rate of consultation for dental care by middle-aged and elderly subjects]. AB - The rate of consultations of elderly and senile patients applying for dental care is analyzed. A total of 510 patients aged 60 years and more who applied to a municipal dentistry center in the southeast region of Moscow were examined. The number of visits to dentist did not depend on age and sex. A notable difference was detected between the rate of consultations for subjects with intact natural teeth and patients who lost natural teeth. PMID- 10368613 TI - [The biomechanical properties of the periodontal tissues]. AB - The literature review of the structure and biomechanical properties of the teeth maxilla segment tissues and also the age and pathologic states dependencies is presented. PMID- 10368614 TI - [Apropos the article "The rapid computerized analysis of dental patients for AIDS" (A. N. Balashov et al., Stomatologiia 1997; 4: 74)]. PMID- 10368616 TI - The new public health: vision for the future. AB - On the fortieth anniversary of the Department of Social and Preventive Medicine, now renamed the Department of Community Health and Psychiatry, the contribution of the Department to the development of public health in Jamaica is briefly reviewed. The paper focuses on the challenges, goals and aspirations for the development of a "new" public health. The challenges include the need for more effective public health leadership, health promotion, better management of the health services and health reform. The latter should provide greater autonomy for the health regions and alternate ways of financing health, and should improve the quality of services provided. The Department is challenged to contribute by establishing a Public Health Residency Programme for doctors leading to a Doctor of Medicine or Doctorate in Public Health; to ensure that epidemiology becomes a basic science in the undergraduate medical curriculum, and that research plays a significant role in postgraduate training of clinical specialists; and to participate more actively in the actual practice of public health. A vision for health in the twenty-first century is given. PMID- 10368615 TI - Forty years. An introduction to the development of a Caribbean public health. AB - The development of public health and primary care in Jamaica is examined with particular reference to the historical events which paved the way for their development: notably, the collaborative work undertaken by the Rockefeller Foundation (Commissions on hookworm, tuberculosis, malaria, yaws); recommendations of the Moyne Commission (leading to the establishment of the West Indies School of Public Health); and the Irvine Commission which recommended the establishment of the University College of the West Indies. A confluence of political, social and international activity in the 1970s proved catalytic in the development of the current ethos of primary health care, and the Department of Social and Preventive Medicine was instrumental in the training of the most innovative addition to the primary care health team, the community health aide. Undergraduate and postgraduate training programmes of the Department are highlighted as it celebrates its fortieth anniversary. PMID- 10368617 TI - Informing Maternal and Child Health (MCH) policy through research. AB - Maternal and Child Health (MCH) policy over the past two decades has been strongly influenced by research. The paper presents examples of some of the research undertaken and its significant influence in shaping health service delivery. Research in child health has focussed on oral rehydration therapy, immunization and perinatal morbidity and mortality. On the maternal side, morbidity and mortality have been examined with particular focus on problems which contribute to maternal and perinatal morbidity and mortality. Policies arising out of the outcome of these studies have influenced organization of service delivery, information system development, manpower development and deployment, maternal education, surveillance/auditing, quality of care, design of physical facilities and selection of equipment. The results of these studies have also led to the identification of areas requiring further study and testing of intervention to correct the deficiencies identified. These studies demonstrate that research can and does influence health policy, and has impacted positively on the quality and cost of care provided through our health services. PMID- 10368619 TI - Pre-hospital emergency medical services, phase I. Role of the Department of Community Health and Psychiatry. AB - The Department of Community Health and Psychiatry was contracted by the Ministry of Health to assist with the implementation of a pilot programme in the Western Health Region to train fire fighters as basic level Emergency Medical Technicians (EMTs), and was responsible for its design, implementation and monitoring for the first 18 months. The course was covered in 440 hours over a 10 week period, and included training in emergency medical response, driving an emergency medical vehicle, emergency medical dispatching and inventory control. Of 76 fire fighters selected for training, 62 graduated, receiving Certificates of Merit from the Ministry of Health, Jamaica and were deployed into service on 17th April 1996. During the period 17 April to 31 December, 1996, the newly trained EMTs responded to 1,299 calls. Medical and surgical cases represented approximately 55% of all calls, followed by trauma (19%), motor vehicle accidents (9%) and obstetric emergencies (8%). Between 15% and 20% of calls in Montego Bay and Negril involved tourists. This first group of EMTs has performed well and was well received by the communities and the persons who used the service. PMID- 10368618 TI - Montserrat. Managing health care in a volcanic crisis. AB - The volcano on Montserrat, after being dormant for over 400 years, has been active for the past two years, last erupting on 27 June, 1997. With the capital, Plymouth, in the unsafe zone, major dislocation of people, facilities and services has occurred. The Health Department is splintered over five sites across an eight mile span and the temporary 30 bed hospital, sited at a primary school, is separated from its Casualty and Out-patient Department and Operating Theatre by 0.25 mile. In order to maintain continuity of care for communities, efforts have been made to keep evacuated clients and their community health workers as close together as possible. The mass emigration has depleted the health services, creating severe stress for those remaining. Elderly relatives have frequently been left behind, necessitating the establishment of special geriatric care facilities to cater to their needs. Increased and continuous health surveillance and mass media education have been integral to the prevention of major disease outbreak--particularly with added challenges to food safety, and management of liquid and solid waste disposal. Cooperation from neighbouring states, particularly Antigua, Barbados and Guadeloupe, as well as from the United Kingdom, has been critical in the management of the continuing crisis. PMID- 10368620 TI - Environmental health. Catching up with the developed world. AB - Environmental health is the ecological balance that must exist between man and his environment in order to ensure his well-being. This paper describes the range of environmental hazards (physical, biological, chemical and social) and discusses the differences in level and quality of environmental health programmes between developed and developing countries. The hazards are the same in both realms; the difference is in the level of advocacy and, thus, the demand for implementation of sound environmental health policies and practices. Issues which developing countries need to address to improve environmental health management are outlined. PMID- 10368621 TI - The social worker. A member of the primary care team? AB - The primary health care team at present does not include social workers as routine members. If however, we, accept the World Health Organization definition of health, which includes social well being, then it follows that the social worker should be considered as a member of the health team to attend to this aspect of health in the service delivery mix. This paper presents the experience of a social worker assigned to the August Town/Hermitage Type III health centre during the period March 1995 to February 1996 and her contribution to patient welfare. The expected roles of the social worker and his or her contribution to the health team are outlined. PMID- 10368623 TI - Alternative medicine: old ideas with new interests? AB - The use of complementary or alternative medicine has greatly increased in developed countries during the last few years. In such countries, laws have been passed and regulations made to protect both the public and the practitioners. There are a growing number of research units in universities, journals, and associations of practitioners of complementary medicine. In Jamaica, too, there is increasing recognition, if not acceptance, of the popular interest in complementary medicine. In preparation for the next century we will need to address some of the legal, educational and scientific issues raised. PMID- 10368622 TI - Integrated primary mental health care. AB - Mental health is increasingly being recognized as contributing significantly to the burden of disease, particularly now that the indicators have shifted from measures of mortality to measures of morbidity. Psychiatric morbidity in the community, based on community surveys, is estimated at 20 to 30% of the population. Increasingly, patients needing mental health services prefer to be in a general health care setting than in specialized centres. Internationally and regionally, the general policy has been to move toward the development of comprehensive mental health programmes integrated within primary health care. This integration may be structural (use of shared facilities), administrative (shared administrative resources) or functional (complete integration of clinical services, with staff jointly responsible for patient welfare). This paper examines how this integration can be achieved, and the potential role of the Department of Community Health and Psychiatry, University of the West Indies in advancing this integrative process through research and training. PMID- 10368624 TI - Improving community care for persons with the acquired immunodeficiency syndrome in Jamaica. AB - A brief review of the HIV/AIDS epidemic in Jamaica is presented along with the details of a pilot programme aimed at training lay persons and practical nurses to assist with home care of persons with AIDS. Current (1997) estimates are that 2 to 4/1,000 Jamaican adults are HIV positive. 2,301 AIDS cases have been reported to the Epidemiology Unit of the Ministry of Health, 55% of whom have died. Males represent 62% of reported cases (1421) and children < or = 9 years old, 7% (170 cases). In 1981 a Family Centre was started at the University Hospital of the West Indies to provide support to persons with AIDS and their families. This initiative identified the need to provide special training to persons who would be responsible for caring the AIDS patient at home, where most patients preferred to be managed. All but three of the 41 persons trained in the pilot programme achieved the required standards of attendance and proficiency. Certificates valid for two years were issued and refresher courses will follow. PMID- 10368625 TI - Chronic diseases: the new epidemic. AB - Mortality statistics show that there has been a significant change in the leading causes of death in Jamaica over the last 50 years, characterized by a decrease in the infectious diseases and those due to undernutrition and an increase in the non-communicable diseases. The various patterns of this epidemiological transition worldwide are outlined and the characteristics of this 'new' epidemic are discussed. Data are presented from the findings of the recent multi-country study of hypertension and diabetes, including Jamaica, which shows that as the body mass index (BMI) increases across the African diaspora, so does the prevalence of hypertension and diabetes. Among the Jamaican population studied, the prevalence of hypertension was 19.1% among males and 28.2% among females. Reported prevalence of previously diagnosed diabetes was 5.3% in men and 10.4% in females. The gender differences are in part explained by the differences in mean BMI which were 23.8 and 27.9, respectively, for males and females. 30.6% of males and 64.7% of females were either overweight or obese, with obesity prevalent in 7.2% of the males and 31.5% of the females studied. The increasing prevalence of obesity across the Caribbean is cause for concern as it significantly impacts on the demand for health and medical care. The identification of these reversible risk factors should be used to inform public policy to tackle what will be a growing concern. PMID- 10368627 TI - Health promotion: issues and answers. AB - In recent years, increased attention has been given to the development of health promotion programmes in a number of countries worldwide. Although health promotion itself is not new, a number of issues have emerged as the underlying concepts are articulated and put into practice. These relate to its relevance and ownership and to practical issues such as measurement of outcomes. This article provides a brief discussion on some of these issues and makes reference to a Caribbean framework for implementing health promotion. PMID- 10368626 TI - Curriculum development in community health: drift or shift? AB - The curriculum in community health is best described as eclectic and dynamic. Its relevance is maintained by its response to the macro-environment; this response, whether innovative or otherwise, may be incremental on the one hand or feature wholesale change consequent on radical rethinking on the other. This paper reviews the content of the emerging curriculum in community health at the University of the West Indies, Jamaica, and attempts to discern the process of change and the factors which have informed these developments. PMID- 10368628 TI - [The ecological connections of dispersal in small mammal populations]. AB - The dispersal of 27 Holarctic species of rodents was quantitatively described on the base of long-term catch method. The relative proportion of non-territorial individuals is an important parameter that reflects the intensity of dispersal. This parameter is statistically related to population (abundance and population type) and species characters (the size of species range and the level of intraspecific differentiation). The contribution of species characters to the variance of migrant portion is lower than contribution of population ones. Isolation and population specialisation are accompanied by the decrease in the portion of disperser, while in pessimal environment the increase in population fluidity is observed. Intensity of dispersal is positively correlated with the size of species range and negatively--with the level of intraspecific differentiation. Thus the dispersal in populations of small mammals serves for colonisation and reparation, providing the stable existence of given group in variable environment. PMID- 10368629 TI - [A shift in prey color preference in the green toad Bufo viridis Laur. after food satiation]. AB - In the behavioural experiments when showing simultaneously coloured stimuli (food targets) and also in choosing between two stimuli, red and blue, it had been shown with a high level of reliability that the hungry Bufo viridis which had not been fed for at least a week, in the overwhelming majority of cases chose the red coloured food targets. Within 1-3 days after an abundant feeding, in the same animals the changes occurred in their preferences of the prey colour. The percentage of choosing the blue targets by the satiated toads significantly increases. In a separate series of experiments had been demonstrated that the changing motivation is conditioned by colour, since hungry and satiated Bufo viridis under conditions of simultaneous demonstration of four stimuli, black and three different grey stimuli, chose exclusively the black stimulus. PMID- 10368630 TI - [The role of behavioral hyperactivation in the formation of anxiety in rodents]. AB - Rodents are a useful tool for studying various stress behaviours, including anxious. Physiological arousal is traditionally thought to accompany anxiety in human and animals. The paper reviews behavioural and pharmacological data on effects of anxiety observed in rodents at different arousal levels. The results indicate that mild arousal might have "paradox" anti-anxiety effects on rodent behaviour while higher arousal level is a pro-anxiety factor. The concept of "pleasurable emotion" produced in rodents by mild arousal is supported. PMID- 10368631 TI - Intravenous injection of an immunoconjugate (anti-PSA-IgG conjugated to 5-fluoro 2'-deoxyuridine) selectively inhibits cell proliferation and induces cell death in human prostate cancer cell tumors grown in nude mice. AB - Current chemotherapeutic and/or endocrine treatments for adenocarcinoma of the prostate are not delivered selectively to prostate cancer cells, therefore, they are used in very high doses that induce many unpleasant side effects in patients. New approaches are, therefore, needed to deliver drugs directly to prostate cancer cells to improve treatment effects. We hypothesized that antibody immunoglobulin G (IgG) against human prostate specific antigen (PSA) (anti-PSA IgG) could function as a carrier protein for conjugated chemotherapeutic drugs (such as 5-fluoro-2'-deoxyuridine, doxorubicin, etc.) and that the immunoconjugate could be delivered selectively to PSA-producing neoplastic prostate. Immunoconjugate would then preferentially inhibit cell proliferation and induce cell death in PSA-producing tumor cells, but not in non-PSA-producing prostate cancer cells or other solid organs of the host. The short-term treatment effect could be assessed by measuring cell death and cell proliferation in tumor bearing animals. We tested our hypothesis by intravenously injecting an immunoconjugate (anti-PSA-IgG-5-fu-2'-d) into nude mice with subcutaneous PSA producing LNCaP or non-PSA-producing Du-145 prostate tumors. During 5 days of treatment, we observed that immunoconjugate was retained preferentially in PSA producing LNCaP tumors where it produced cytotoxic effects in neoplastic prostate cells as revealed by decreased cell proliferation and increased cell death, but similar effects were not observed in non-PSA-producing Du-145 tumor cells or mouse organs. Analysis of untreated control mouse with LNCaP tumor, anti-PSA-IgG alone, anti-irrelevant-IgG-drug complex, and drug alone treatments indicated that there was little or no cytotoxic effects of these treatments on LNCaP and Du-145 tumors, and host organs. Our analysis of control and experimental data showed that the immunoconjugate was highly specific in imparting cytotoxic effects on LNCaP prostate tumors, but not on Du-145 tumors and mouse organs. Thus, we have shown that the immunoconjugate selectively delivered a chemotherapeutic drug to PSA-producing prostate tumor cells where it produced measurable cytotoxic effects on cell proliferation and cell death. This is the first report to show a successful delivery of a chemotherapeutic drug in the immunoconjugate to PSA producing LNCaP prostate tumors in nude mice and without inducing cytotoxic effects on mouse organs. PMID- 10368632 TI - Differential induction of premature chromosome condensation by calyculin A in human fibroblast and tumor cell lines. AB - The feasibility of chemically-induced premature chromosome condensation (PCC) was tested in two human fibroblast and two human malignant melanoma cell lines. To induce PCCs, calyculin A, a potent protein phosphatase inhibitor, was used at a final concentration of 80 nM. Attached cells and cells put into suspension by trypsinization were incubated with calyculin A at 37 degrees C for 1 hour. Calyculin A was able to induce PCCs in all the phases of the cell cycle with the tumour cell lines giving the highest PCC frequency. No systematic differences were observed between attached cells and cells put into suspension by trypsinization. However, a cytotoxic effect that led to the loss of 50% to 80% of the treated cells was observed. The cytotoxic effect was more severe in the fibroblast than in the tumour cell lines. The appearance of deformed, fragile and fragmented nuclei with no particular chromatin condensation would explain to some extent this cytotoxic effect. A better understanding of the mechanism of action of calyculin A would help generalizing its use to study interphase chromosome aberrations. PMID- 10368634 TI - Growth inhibition of nasopharyngeal carcinoma cells by EGF receptor tyrosine kinase inhibitors. AB - Nasopharyngeal carcinoma (NPC) is a malignancy of epithelial origin occurring with a high incidence in southern China and southeast Asia. Radiotherapy is the main treatment modality for NPC. No effective chemotherapy is available. Since prevention of EGF/EGFR binding by an EGFR specific monoclonal antibody suppressed the growth of NPC xenografts, we examined potential anti-NPC activity by a group of specific inhibitors of the EGFR family of tyrosine kinases. We found that HONE T1 NPC cells expressed high levels of EGFR tyrosine kinase activity upon stimulation by EGF. The receptor tyrosine kinase activity was specifically inhibited by either reversible (PD158780) or irreversible (PD168393) inhibitors specific for EGFR family tyrosine kinases. This inhibition led to a dose dependent suppression of anchorage-independent growth as determined by soft agar assays. A structural analog (PD159805) with no inhibitory activity against EGFR tyrosine kinase had no effect on HONE-T1 cell growth in agar. Furthermore, growth of HONE-T1 xenografts in SCID mice was also inhibited by treatment with PD158780 and PD 168393. This data provides an appealing application of EGFR tyrosine kinase inhibitors for the treatment of nasopharyngeal carcinomas. PMID- 10368635 TI - Detection of chromosome 11q13 amplification in head and neck cancer using fluorescence in situ hybridization. AB - BACKGROUND: Head and neck squamous cell carcinoma (HNSCC) remains a cancer with one of the lowest five-year survival rates. Despite a better understanding of the disease and recent advances in diagnosis and treatment, survival rates for HNSCC patients have not improved. Chromosomal abnormalities are common in HNSCC, and aberrations of chromosome 11q13 have been correlated with a poor prognosis. MATERIALS AND METHODS: In this study we utilized fluorescence in situ hybridization (FISH) to determine the incidence of 11q13 amplification in twenty primary HNSCC tumors. INT-2 was used as the 11q13 probe, and 9 and 11 centromeric probes were used as controls. RESULTS: Polysomy, greater than two copies of chromosome 11, was found in 2 of 20 tumors. INT2 (11q13) amplification was found in 3 other tumors. CONCLUSIONS: These preliminary studies indicate tht analysis of a larger sample of tumors using FISH may yield important diagnostic and prognostic information about head and neck tumors. PMID- 10368633 TI - In vitro chemopreventive efficacy screening using human keratinocytes and the in vivo data correlation. AB - Agents with potential cancer preventive activity were screened for efficacy in the Human Epidermal Cell (HEC) Assay. The HEC Assay measures inhibition of propane sultone-induced changes in the growth and/or differentiation in early passage keratinocyte cultures. The assay biomarkers were calcium tolerance, growth inhibition, and involucrin induction. The HEC Assay also provides information on the cytotoxicity of the agents following acute and chronic exposure. Agents were evaluated at non-toxic doses in the HEC Assay. The HEC Assay has been used to screen twenty-eight agents for chemopreventive efficacy. A positive response in one or more endpoints of the HEC Assay correlates 100% (16/16) with a positive response in one or more of the animal cancer prevention models (J. Cell. Biochem., 26S:29-53, 1996). The overall sensitivity for predicting efficacy in animals is 84%. The available data suggest that a positive assay response appears to be highly predictive of efficacy in vivo. PMID- 10368637 TI - Doxorubicin-induced cell death in highly invasive human gliomas. AB - Glioblastoma is the most invasive form of primary brain tumors, and is often refractory to chemotherapy. Herein, we provide evidence that two highly invasive human glioma cell lines U-87 MG and U-373 MG, entered apoptosis after 48 hours following 24 h growth arrest induced by Doxorubicin (10 micrograms/2 x 10(5) cells/ml). Apoptosis depended solely on the level of intracellular drug accumulation, and it was not related to a functional p53 tumor suppressor factor. The multidrug resistance gene 1 (mdr-1) encoded P-glycoprotein (P-gp) was weakly expressed in these cells upon exposure to Doxorubicin, and exerted no influence on the extent of cellular drug efflux. Drug efflux occurred only in U-373 MG glioma cells subsequent to physical damage of the membrane upon exposure to Doxorubicin. Pretreatment of tumor cells with 10 micrograms/ml Doxorubicin precluded tumor formation on the chorioallantoic membrane (CAM) of embryonated hen eggs. Single-dose application of 0.4 microgram Doxorubicin on CAM/U-87 MG and CAM/U-373 MG tumor transplants inhibited tumor invasion in CAM tissue by 40 to 50%. These data suggest that highly invasive glioblastomas can be driven to apoptosis following growth arrest induced by Doxorubicin, providing that intracellular drug accumulation suffices cytotoxic levels. PMID- 10368636 TI - Immunocytochemical detection of hMSH2 and hMLH1 expression in oral SCC. AB - BACKGROUND: The loss of DNA mismatch repair system was reported in hereditary non poliposis colon cancer and in other tumours. The aim of this study was to detect the protein expression pattern of hMSH2 and hMLH1 in oral squamous cell carcinoma (SCC) by immunohistochemistry in paraffin-embedded tissues. MATERIALS AND METHODS: 5 specimens, obtained from healthy oral mucosa, and 20 from oral SCC were tested with anti-hMSH2 and anti-hMLH1 monoclonal antibodies. RESULTS: Six cases (30%) showed nuclear positivity in differentiated areas (G1) and cytoplasmic positivity in areas with a lower degree of differentiation, four cases (20%) showed only cytoplasmic positivity, and only one (5%) no staining. One case of oral SCC (5%) showed no hMLH1 staining in the tumoral cells, even if normal squamous epithelium available in this section showed a nuclear positivity; six cases (30%) showed nuclear positivity in differentiated areas (G1) and cytoplasmic positivity in areas with a lower degree of differentiation, three cases (15%) showed only cytoplasmic positivity. CONCLUSIONS: These data suggest that examination of hMSH2 and hMLH1 protein expression by immunohistochemistry is important in oral SCC. The analysis of mismatches expression in these cases of oral SCC might suggest that an absent nuclear staining for both hMSH2 and hML1 could constitute a hallmark of potential phenotype mutator for this type of neoplasia. PMID- 10368638 TI - Detection of p53 protein accumulation in sputum and lung adenocarcinoma associated with indoor exposure to unvented coal smoke in China. AB - Lung cancer in Xuan Wei (XW), China has been linked to exposure to unvented coal smoke and adenocarcinoma, especially bronchioloalveolar carcinoma, is most common. p53 mutations occur commonly in lung cancers and usually generate detectable levels of p53 protein accumulation. Sputum is noninvasive to collect and ideal for screening p53 abnormalities. p53 protein accumulation was detected by immunohistochemistry in lung tumors and sputa from XW lung cancer patients to determine (1) the role of p53 in lung pathogenesis, and (2) feasibility of detecting p53 protein accumulation in sputum, p53 protein accumulation was detected in 73% (22/30) of lung adenocarcinomas from XW females exposed to coal emissions and significantly higher than the control cases (33%, p < 0.05). In sputum, we detected p53 overexpression in tumor cells in 54% (13/24) of XW cases and also in dysplastic cells (50% or 4/8). These findings suggest that p53 abnormalities is important in XW lung cancer etiology. PMID- 10368640 TI - Treatment of metastases of solid mouse tumours by NAMI-A: comparison with cisplatin, cyclophosphamide and dacarbazine. AB - The effects of NAMI-A, a novel ruthenium compound endowed with selective antimetastatic action, were tested on solid metastasising tumours of the mouse in comparison to cisplatin, cyclophosphamide and dacarbazine. Each compound was administered i.p. as freshly prepared solutions in isotonic saline in combination with surgical removal of primary tumour, and was used at the maximum tolerated dose. NAMI-A significantly reduced the growth of lung metastases either when given prior to surgery (early growing tumours) on TS/A adenocarcinoma or after surgical ablation of primary tumours (already established lung metasases) on Lewis lung carcinoma. The postsurgical treatment of mice bearing MCa mammary carcinoma caused a significant prolongation of the life-span of the treated animals. In the comparison experiments, dacarbazine was completely ineffective, cisplatin was as active as NAMI-A on MCa mammary carcinoma, slightly less active than NAMI-A on TS/A adenocarcinoma and inactive on Lewis lung carcinoma, and cyclophosphamide was always more active than any other treatment performed. These data stress that NAMI-A, independently of the lack of direct cell cytotoxicity, when compared to the reference drugs, has a potent therapeutic effect in mice bearing solid metastasising tumours. PMID- 10368642 TI - 5-Fluorouracil simultaneously maintains methotrexate antineoplastic activity in human breast cancer and protects against methotrexate cytotoxicity in human bone marrow. AB - High-dose methotrexate (MTX) cytotoxicity is maintained in MCF-7 breast cancer cells but reduced in Hs824.T human bone marrow by a priming and nontoxic 5 fluorouracil (5-FU) dose. When MCF-7 breast or Hs824.T bone marrow cells are incubated with 10 microM 5-FU and 10 microM MTX for 48 h, the growth rates of breast cancer cells were 97.59 +/- 0.97% and 21.81 +/- 3.33% of the control rate, respectively, and the growth rates of bone marrow cells were 90.61 +/- 3.71% and 29.58 +/- 2.99% of the control rate. The combinations of 5-FU 2 h prior to MTX or MTX 2 h prior to 5-FU followed by a 48 h incubation, respectively, gave growth rates of 20.96 +/- 2.44% and 19.86 +/- 2.56% of the control rate for MCF-7 cells. In bone marrow cells, the combinations of 5-FU 2 h prior to MTX or MTX 2 h prior to 5-FU followed by a 48 h incubation, respectively, gave growth rates of 79.66 +/- 7.41% (protection) and 31.39 +/- 1.77% of the control rate. Similar patterns to bone marrow emerges in platelets. These studies suggest that: a) MTX and 5-FU combination on the growth of human MCF-7 breast cancer cells is independent of sequence; and b) a priming-dose of 5-FU will protect bone marrow from MTX cytotoxicity but not breast cancer cells. Therefore, a priming and non-toxic dose of 5-FU and MTX may have maximum antineoplastic activity while at the same time provide protection to the hematopoietic system. PMID- 10368639 TI - Novel small molecule alpha v integrin antagonists: comparative anti-cancer efficacy with known angiogenesis inhibitors. AB - Recent evidence supports the involvement of integrins in angiogenesis: blockade of alpha v beta 3 and alpha v beta 5 integrins disrupts angiogenesis leading to decreased blood vessel formation and hence decreased tumor growth. We hypothesized that av antagonists could inhibit tumor growth in tumor cells devoid of alpha v beta 3 integrins. We evaluated SM256 and SD983, novel small molecules that are specific av antagonists in mouse models of angiogenesis and tumorigenesis, and compared them with standards: TNP470, a fumagillin analog in the clinic, and flavopiridol, a cell cycle kinase inhibitor. In vitro SM256 was a selective alpha v beta 3 inhibitor with an IC50 = 4nM, and the affinity of SD983 against the mouse endothelial alpha v beta 3 integrin yielded an IC50 = 2nM and an IC50 = 54nM against alpha v beta 5. In the mouse Matrigel model of angiogenesis SM256 decreased blood vessel formation (hemoglobin content) with an ED50 = 0.055 ug/kg/day, tenfold more potent than TNP470. SG545, an ester of SD983, decreased blood vessel formation with an ED50 = 6 ug/kg/day, while flavopiridol ED50 = 18 ug/kg/day. In the mouse xenograft model, using human colon carcinoma RKO cells that do not express alpha v beta 3 but express alpha v beta 5, tumor growth was inhibited by SG545 (10 mg/kg/day) and flavopiridol (5 mg/kg/every other day) 40% and 70%, respectively (p < 0.05). Although the proliferative index (measured by BrdU incorporation) was not significantly changed with SM256, SG545 or flavopiridol (29-32%), the apoptotic index increased significantly (p < 0.05) in the SM256 and SG545-treated groups (2.3-2.7%) compared with controls (1.1%), suggesting increased cell death contributed to decreased tumor volumes. Neovascularization decreased with SM256 and SG545 treatment. The data demonstrate that potent selective av antagonist can target endothelial cells, tumor cells, inhibit angiogenesis and inhibit tumor growth. PMID- 10368641 TI - Mechanisms of cell transformation by Herpesvirus saimiri. AB - Herpesvirus saimiri is a virus capable of inducing oncogenic transformation of T lymphocytes of New World primates and immortalizing human T cells in vitro. T lymphocytes immortalized by H. saimiri demonstrate functional biological responses to their antigens. Therefore, H saimiri-induced transformation of T cells emerges as a very powerful tool of T-cell biology. The mechanism of this transformation remains to be elucidated. However, it is apparent that H. saimiri proteins Tip and StpC are crucial for T-cell transformation. Potential mechanisms mediating the effects of Tip and StpC, as well as the possible role of other H. saimiri proteins, are discussed in this review. PMID- 10368643 TI - Electrically mediated drug delivery for treating subcutaneous and orthotopic pancreatic adenocarcinoma in a hamster model. AB - BACKGROUND: Current therapies for pancreatic adenocarcinoma only benefit a fraction of those diagnosed with this disease. New strategies for improving treatment are clearly needed. This study investigated the use of electrically mediated drug delivery for the treatment of pancreatic adenocarcinoma in a hamster model. MATERIALS AND METHODS: Hamster PC-1 pancreatic adenocarcinoma cells and Golden Syrian hamsters were used as a model. RESULTS: In vitro testing indicated that bleomycin was more effective than Cisplatin and Doxorubicin when delivered using pulsed electric fields. Treatment of subcutaneous tumors with bleomycin and electric fields resulted in a 100 percent complete response rate. No effect was observed when either drug or pulses were used alone. Treatment of tumors induced in the gland resulted in a 25 percent complete response rate. CONCLUSIONS: Electrochemotherapy was highly effective for subcutaneous tumors. There was also a significant antitumor effect for the more complex and clinically relevant intraoperative treatment of tumors in the pancreas. PMID- 10368644 TI - Schedule-dependent efficiency of thermochemotherapy in vitro with etoposide and heating at 43 degrees C. AB - We have investigated the effect of hyperthermia (HT) treatment on the extent of etoposide-induced apoptosis in human leukemia HL-60 cells. The cells were heated at 43 degrees C for 15 to 60 min. Relative quantitative changes in the apoptotic (Ap) fractions were monitored by flow cytometry, HT treatment for 15 min prior to or concurrently with etoposide exposure had no effect on the Ap fraction generated by the drug alone, whereas, 60 min heating resulted in an increase in the Ap fraction. Heating of the cells at 6 to 24 hr after drug exposure enhanced the Ap fraction. Taken collectively, the results indicate that appropriately scheduled HT and etoposide treatments may lead to a thermochemotherapy protocol that will result in increased etoposide-induced death of human leukemia cells. PMID- 10368645 TI - Chemopreventive activity of a C-glucuronide analog of N-(4-hydroxyphenyl) retinamide-O-glucuronide against mammary tumor development and growth. AB - The long term chemopreventive effects of the N-(4-hydroxyphenyl) retinamide-O glucuronide (4-HPROG), and its stable C-linked benzyl glucuronide analog, retinamidobenzyl glucuronide (4-HPRCG) on the growth and development of 7,12 dimethylbenz[a]anthracene-induced mammary tumors were compared. The retinamidobenzyl glucuronide is stable toward acid hydrolysis and resists the actions of beta-glucuronidase. The results indicate that the C-linked glucuronide analog, 4-HPRCG has a greater chemopreventive potency than an equimolar concentration of 4-HPROG. Tumor latency was 15% longer in rats fed 2 mmol/kg diet of 4-HPRCG as compared to 4-HPROG. At 80 days post DMBA-intubation, tumor incidence was 57% and 27% in the 4-HPROG and 4-HPRCG treated rats, respectively. Tumor multiplicity was also markedly decreased in the 4-HPRCG treated rats. At 80 days post DMBA intubation the control rats had an average of 1.43 tumors/rat compared to 0.71 and 0.36 tumors/rat in the 4-HPROG and 4-HPRCG respectively. The higher potency and low toxicity of 4-HPRCG suggest that this stable analog may have an in vivo chemopreventive advantage over its analog, 4-HPROG. The results also demonstrated that these glucuronide analogs do not bind effectively in vitro either to the nuclear retinoid receptors or to the cellular retinoid binding proteins. Regardless of the mode of action of these retinoids, they are clearly effective chemopreventive agents. PMID- 10368646 TI - Histochemical and morphological identification of Kupffer cells activated by cisplatin. AB - Cisplatin is a potent anti-cancer agent which has been shown to activate Kupffer cells. These activated macrophages demonstrate an increase in extensions, lysosomes, and peroxisomes increasing their anti-tumor activity. Wistar rats were treated with cisplatin (9 mg/kg) and sections of liver were excised for light and electron microscopic analysis at 1, 6, 15, and thirty days post treatment. Non specific esterase staining was used to differentiate Kupffer cells using light microscopy, morphologic criteria were used for TEM analysis. Liver sections taken 6 days post treatment showed the greatest number of activated macrophages, with the highest degree of activation. Interaction between natural killer cells and Kupffer cells was only seen 6 days post treatment. These results show that cisplatin's ability to enhance the immune system requires several days post treatment to reach maximum potency. PMID- 10368647 TI - Oncostatin M (OM) promotes the growth of DU 145 human prostate cancer cells, but not PC-3 or LNCaP, through the signaling of the OM specific receptor. AB - BACKGROUND: Oncostain M (OM) is a member of the interleukin-6 (IL-6) cytokine family and all members utilize gp130 as a common signal transducing receptor. Tyrosine phosphorylation of the transcription factor STAT3 plays an important role in IL-6 cytokine family-mediated reactions. OM signals are transduced through two different OM receptor complexes: a leukemia inhibitory factor (LIF)/OM receptor (a heterodimer of LIFR beta and gp130) and an OM specific receptor (a heterodimer of OMR beta and gp130). This study examined the expression of these two OM receptors as well as the effect of OM on the growth of prostate carcinoma cell lines. MATERIALS AND METHODS: PC-3, DU 145 and LNCaP cells were used. The expression of OMR beta and LIFR beta was determined by RT PCR. Tyrosine phosphorylation of STAT3 was analyzed by Western blotting. RESULTS: OM promoted the growth of DU 145 but not that of PC-3 and LNCaP. LIF had no effect on the growth of any of these cell lines. DU 145 and PC-3 expressed much higher levels of OMR beta mRNA than those of LIFR beta mRNA. Neither OMR beta nor LIFR beta mRNA was detected in LNCaP. OM, but not LIF, induced rapid tyrosine phosphorylation of STAT3 in DU 145. PC-3 lacked STAT3 expression. CONCLUSIONS: OM has been known to be a cytostatic cytokine for tumor cells. Conversely, we show that OM promotes the growth of DU 145. Further, our findings suggest that the growth-promoting signals of OM is mediated through the OM specific receptor with subsequent activation of STAT3. PMID- 10368650 TI - The activity of unloaded gelatin nanoparticles on murine melanoma B16-F0 growth in vivo. AB - BACKGROUND: Bacillus Calmette-Guerin (BCG) vaccine is a non-specific immunostimulant which has been used clinically in the treatment of melanoma. In this communication, the antimelanoma activity of BCG was related to its fibronectin-binding properties and mimicked using gelatin nanoparticles. MATERIALS AND METHODS: The fibronectin-binding properties of aqueous gelatin solutions, gelatin nanoparticles, BCG vaccine, and PS1 (a glucan extracted from Tice BCG) were compared by an enzyme-linked immunosorbent assay and their ability to suppress murine B16-F0 melanoma in vivo investigated. RESULTS: Aqueous gelatin solutions, gelatin nanoparticles and BCG all bound to fibronectin in vitro. The immunostimulant PS1 did not. In vivo, BCG and gelatin nanoparticles suppressed melanoma growth while PS1 and aqueous gelatin solutions had no effect. CONCLUSIONS: The antimelanoma activity of BCG is not due to the associated immunostimulatory glucan but can be correlated to its fibronectin-binding properties. Since solutions of gelatin have no effect whereas nanoparticles produce total suppression, this suggests a relationship between the volume of the fibronectin-binding entities and their antitumour activity. Thus, gelatin nanoparticles may represent an attractive alternative to the use of BCG vaccine in melanoma treatment. PMID- 10368649 TI - Altered sensitivity of deoxyadenosine-resistant mouse leukemia L1210 cells to various kinase inhibitors. AB - The deoxyadenosine-resistant mouse leukemia L1210 cell line (Y8) has previously been shown to have phenotypic differences that appear unrelated to the altered properties observed at the level of ribonucleotide reductase (RR). In response to various stress factors, the parental wild-type (WT) L1210 cell line undergoes cell cycle arrest; Y8 cells become apoptotic. These responses are p53 independent. Cell cycle regulation also appears different between the two cell lines, suggesting that Y8 cells are more apoptotic because of alterations in their cell cycle compared to WT cells. In order to study the relationships between cell cycle regulation and apoptosis, the effects of 2-aminopurine (2-AP), wortmannin, and PD98059, were studied on WT and Y8 cells. 2-AP induced G2/M block in both WT and Y8 cells with differences in G0/G1 and S phase contents between the two cell lines. Wortmannin induced G0/G1 block in Y8 cells, while exhibiting no effect on WT cells. PD98059 had no effect on the cell cycle of either WT or Y8 cells. In response to each inhibitor, Y8 cells underwent apoptosis to a much greater extent than the parental WT cell line. These data suggest that the specific pathways that converge on the cell cycle are altered and may be involved in the differences between a tumor cell to block in cell cycle or to undergo apoptosis. PMID- 10368648 TI - Screening of potential cancer preventing chemicals as aromatase inhibitors in an in vitro assay. AB - The inhibition of human placental aromatase was used to rank a series of compounds, with the objective of selecting compounds for further evaluation as chemopreventive agents. (+/-)-p-Aminoglutethimide, introduced over two decades ago as a treatment for breast cancer, had an IC50 of 6.5 microM. Five compounds were more potent than aminoglutethimide in this assay: (+)- vorozole, 4 hydroxyandrostenedione, miconazole nitrate, plomestane, and 4-methoxy-androst-4 ene-3,17-dione. Other compounds with known chemoprevention activity, such as curcumin and genistein, were inactive. This assay for aromatase inhibitors is a rapid, economical way of ranking compounds for further development as chemoprevention agents. PMID- 10368651 TI - Antioxidant activity of michellamine alkaloids. AB - The alkaloids michellamines A, B, and C are natural products isolated from a Central African tropical plant Ancistrocladus korupensis. We have investigated the radical scavenging ability of these compounds. The alkaloids inhibited the azo-induced oxidation of beta-phycoerythrin with IC50 values in the 0.5- to 0.8 microM range. Michellamine B also protected rat liver mitochondria against lipid peroxidation induced by adenosine diphosphate and Fe2+. The alkaloids were more potent antioxidants in these assays than several compounds being considered clinically as chemoprevention agents. PMID- 10368652 TI - Thionins from Viscum album L: influence of the viscotoxins on the activation of granulocytes. AB - BACKGROUND: Extracts from European mistletoe (Viscum album L.) are applied in adjuvant cancer treatment, and some components, especially the mistletoe lectins (ML) have been immunologically characterised, but not the thionins, termed viscotoxins (VT). MATERIALS AND METHODS: The influence of the VT on human granulocytes was studied by flow cytometry: E.coli co-stimulated respiratory burst by oxidation of dihydrorhodamine 123 to rhodamine 123 and phagocytosis by ingestion of FITC-labelled E.coli. RESULTS: VT (25 and 250 micrograms/ml), in contrast to ML, significantly enhanced phagocytosis and burst activity. VT-rich mistletoe extracts also exerted significant effects. In addition, E.coli activated granulocytes positively stain with Annexin-V and propidium iodide only due to 250 micrograms/ml VT incubation, suggesting that at this concentration burst activity was induced by the physiological activity of granulocytes after microbial ingestion and also by cytotoxic effects. CONCLUSION: Viscotoxins exert yet unknown strong immunomodulatory effects on human granulocytes, which might be of benefit for tumour patients, in addition to their cytotoxic properties. PMID- 10368653 TI - Immunohistochemical and molecular analysis of bax, bcl-2 and p53 genes in laryngeal squamous cell carcinomas. AB - BACKGROUND: We investigated the role of apoptosis and its potential alterations in laryngeal squamous cell carcinomas (LSCCs) by evaluating bax, bcl-2 and p53 protein expression in 50 cases and by characterising the molecular status of the bax and p53 genes. MATERIAL AND METHODS: p53 and bax gene mutations were investigated by means of PCR/SSCP and direct DNA sequencing, and bax, bcl-2 and p53 protein expression by means of immunohistochemistry. RESULTS: We identified p53 gene mutations in 17/50 cases (34%); p53 expression in 26 of the 50 cases (52%); bcl-2 expression in 5/50 cases (10%); bax expression in 32/47 cases (68%). 18/33 cases with a wild type p53 gene overexpressed p53 protein: 12 cases (approximately 66%) were bax+/bcl-2-. Of the remaining cases without p53 protein expression, seven cases (approximately 47%) were bax+/bcl-2-. CONCLUSIONS: Our observations suggest that the overexpression of p53 may contribute to the repression of bcl-2 and the induction of bax expression in LSCCs. However, the fact that a number of cases not expressing p53 did not present any clear up regulation of bax or down-regulation of bcl-2 suggests that bcl-2 and bax may be regulated by various mechanisms other than p53. PMID- 10368654 TI - Combinations of P-glycoprotein blockers, verapamil, PSC833, and cremophor act differently on the multidrug resistance associated protein (MRP) and on P glycoprotein (Pgp). AB - Clinical studies are currently in progress to evaluate functional modifiers of P glycoprotein (Pgp), an efflux pump associated with resistance to cancer chemotherapy. However, the effects of these modifiers on a more recently discovered efflux pump, the multidrug resistance associated protein (MRP), have not yet been fully characterized. MRP is expressed in most human tissues and is overexpressed in several tumor types. For these reasons, we have investigated the effects of three prototype Pgp modifiers, which act by different modes on the function of Pgp, on the function of MRP in two MRP-overexpressing cell lines: UMCC/VP lung and MCF-7/VP breast cancer cells. Clinically optimal plasma levels of verapamil, cremophor, and PSC833 have been shown to completely block the function of Pgp in Pgp-over expressing cells. However, in the two MRP-over expressing cell lines, these modifiers only partially blocked the function of MRP and combinations of these optimal concentrations acted antagonistically. Similar antagonistic effects were seen with combinations of suboptimal concentration levels of these blockers, while these combinations resulted in synergistic effects in Pgp overexpressing cells. For two biophysical parameters measured at the plasma membrane, membrane fluidity and membrane potential, the effects of these modifiers were essentially similar in Pgp and MRP expressing cells. We suggest that the 170 kD Pgp and the 190 kD MRP glycoproteins, imbedded in the plasma membranes, respond differently to simultaneous effects of the investigated prototype resistance modifiers. These results also suggest that the identification of the specific mechanism of drug resistance is important for the selection of chemotherapeutic strategies to block the efflux pump on the cancer cell. PMID- 10368655 TI - Local inflammation may influence oral tumor cell differentiation. AB - The mRNA levels of keratin K13, involucrin, protein kinase C alpha and epsilon, and interferon-gamma and its receptors were examined in biopsies from human oral squamous cell carcinomas. Expression of all the genes was elevated in the histologically more differentiated tumors, but it was at or below normal (perilesional control) levels in the poorly differentiated ones. For the same set of biopsies, we had previously shown that the well differentiated tumors expressed higher levels of T cell markers. As interferon-gamma stimulates differentiation, its secretion by inflammatory cells at the tumor site may influence the differentiation status of the tumor. PMID- 10368657 TI - Down-regulation of IL-10 enhances the efficacy of locoregional immunotherapy using OK-432 against malignant effusion. AB - To determine the significance of interleukin (IL)-10 in antitumor immune response, the effect of the down-regulation of tumor-derived IL-10 on locoregional immunotherapy was investigated. C3H/HeN mice were intraperitoneally (i.p.) inoculated with IL-10-producing murine breast cancer cell line, FM3A, and treated with locoregional administration of OK-432 with or without anti-IL-10 monoclonal antibody (mAb). Anti-IL-10 mAb did not affect the in vitro growth of FM3A cells. Administration of OK-432 plus anti-IL-10 mAb remarkably delayed the retention of malignant ascites and prolonged the survival of mice compared with the administration of OK-432 alone. Spleen cells which were collected from mice treated with OK-432 plus anti-IL-10 mAb and further stimulated in vitro with inactivated FM3A cells exhibited significantly higher cytotoxicity against FM3A cells than those from mice treated with OK-432 alone or from the control mice. The expression of major histocompatibility complex (MHC) class II molecules on spleen cells was up-regulated in vitro by the addition of OK-432 and anti-IL-10 mAb. Using semi-quantitative reverse transcription-polymerase chain reaction (RT PCR), cytokine mRNA levels of peritoneal exudate cells (PEC) and spleen cells were assessed on day 7 (before treatment) and day 14 (after treatment). In PEC, increased expression of IL-2 was observed with the administration of OK-432 plus anti-IL-10 mAb. In spleen cells, the expression of IL-2, IL-12 and IFN-gamma were strongly induced, and IL-4 expression was reduced by the administration of OK-432 plus anti-IL-10 mAb. It is suggested that down-regulation of tumor-derived IL-10 induces the up-regulation of the T helper type (Th) 1 population, resulting in an enhancement of the efficacy of locoregional immunotherapy with OK-432. PMID- 10368658 TI - Lecithinized ascorbic acid (PC-AS) effectively inhibits murine pulmonary metastasis. AB - In order to enhance the lipophilicity and develop the efficacy of ascorbic acid (ASA), we synthesized lecithinized ascorbic acid (PC-AS), in which a lecithin was covalently bound to ASA. Its pharmacological activity was also evaluated. The IC50 value of scavenge superoxide anions generated from hypoxanthine in combination with xanthine oxidase, indicated that the antioxidative activity of PC-AS (IC50; 22.19 microM) was about 60% of that shown by ASA (IC50; 13.35 microM). Also, PC-AS suppressed in vitro cell growth of Meth A-T, a highly metastatic cell line established by us. Although its potency (IC50; 110.0 microM) was a little lower than that of ASA, dramatic suppression was observed under serum-free culture conditions (IC50; 13.0 microM). In addition, N-acetylcysteine (NAC), an antioxidant, showed an additive inhibitory effect on cell growth in combination with PC-AS and ASA. Biodistribution studies revealed that PC-AS persisted longer in the blood (AUC0-240 min; 182.8 nmole min ml-1) than ASA (AUC0 240 min; 79.35 nmole min ml-1). It should be noted that intravenous preadministration of PC-AS significantly and dose-dependently reduced the number of colony formation in an experimental murine pulmonary metastasis model. ASA had little effect. [3H]-labeled Meth A-T cells predominantly accumulated in the lung, metastatic target organ, which was reduced by PC-AS. Our in vivo study showed that PC-AS could not totally prevent pulmonary invasion of Meth A-T cells, however, PC-AS effectively inhibited the number of metastatic colony formation. PC-AS's potency was superior to that of unmodified ASA. These findings might be in part ascribed to changes to lecithinization-induced biodistribution, antioxidative activity and cytotoxicity. PMID- 10368656 TI - Vitamin D3 analogue KH1060 combined with TPA synergistically induces mature macrophages in human myeloblastic leukemia ML-1 cells. AB - A human myeloblastic leukemia cell line, ML-1, was induced to differentiate along the monocytic lineage following exposure to a 1,25-dihydroxyvitamin D3 analogue, 20-epi-22-oxa-24a,26a,27a-tri-homo-1,25-dihydroxyvitam in D3 (KH1060) or 12-O tetradecanoylphorbol-13-acetate (TPA). The combination of KH1060 and TPA synergistically induced differentiation of ML-1 cells into mature macrophages with multinuclei. Maturation was also observed in other differentiation characteristics such as phagocytic activity, a-naphthyl acetate esterase activity, and expression of surface antigen CD14. Differentiated ML-1 cells showed attenuation of telomerase activity and cessation of proliferating activity based on evaluation of the expression of genes related to cell growth potential. Remarkable synergism was observed in TNF production. Treatment with TPA prior to KH1060 resulted in only slight production of TNF; however, treatment with KH1060 preceding TPA induced a substantial amount of TNF. PMID- 10368659 TI - Histone H1 kinase activity in one-cell mouse embryos blocked in the G2 phase by X irradiation. AB - The activation of the p34cdc2/cyclin B complex is responsible for driving the cell cycle from the G2- to the M-phase. To investigate the effects of irradiation on the activity of the p34cdc2/cyclin B complex in preimplantation embryos, we irradiated one-cell mouse embryos with 2.5 Gy of X-rays at the early pronuclear stage, and measured the fluctuations of histone H1 kinase activity (a biochemical indicator of the kinase activity of the p34cdc2) at different times during the radiation-induced G2-arrest. BALB/c embryos were chosen for these experiments, since earlier results obtained in our laboratory had shown that such a treatment induces a G2-arrest of about 20 hours in more than 90% of the embryos. Our data showed that histone H1 kinase activity of irradiated embryos remained at a very low level during the period of G2-arrest. The level of activity found during late division of the G2-arrested embryos was also significantly lower in comparison with that of control embryos or irradiated embryos dividing without delay. All together, our results suggest that a) low levels of histone H1 kinase activity are sufficient for the division of one-cell embryos, b) there could be a link between the levels of histone H1 kinase activity in mitosis and the health status of the embryo. PMID- 10368660 TI - Transport of doxorubicin in mast cell granules and the effect of the calcium antagonist verapamil. AB - P-glycoprotein has been identified in mast cells stabilized in culture as well as in rat peritoneal mast cells, and is primarily concentrated on the granular membrane. This study aimed to define the role of this protein in the transport and accumulation of doxorubicin in mast cell granules and in its histamine releasing effect. The reverting agent verapamil, that is a substrate for P glycoprotein, inhibited doxorubicin uptake in intact mast cells in a dose and time dependent manner, but had no effect on the exocytotic action of the antineoplastic drug. Doxorubicin was also concentrated in granules with intact membranes and the uptake was dependent on temperature and showed a trend for saturation. Verapamil and vinblastine, another substrate for P-glycoprotein, significantly reduced doxorubicin concentrations in intact granules. Similar results were obtained with the metabolic inhibitors sodium metavanadate, N ethylmaleimide, and sodium azide, whereas ouabain, an inhibitor of sodium potassium ATPase, was without effect. Doxorubicin was taken also up in granule remnants, consisting of a proteoglycan matrix without membrane, that are extruded from mast cells upon stimulation. However, the uptake was not dependent on temperature and was not modified by P-glycoprotein substrates or metabolic inhibitors. Rat peritoneal mast cells were examined for the expression of P glycoprotein at the protein level with C219 monoclonal antibody, using Western blot, confirming that P-glycoprotein was expressed in mast cells. These data suggest the presence of a P-glycoprotein active in the transport of doxorubicin, in mast cell granules. PMID- 10368661 TI - Molecular genetic analysis of primary lung cancer and cancer metastatic to the lung. AB - The lung is a very common site for primary cancer and metastatic disease. Advances in tumor biology have provided insight into the sequence of genetic alterations leading to tumor and metastasis formation in the lung. In this review we address two genetic alterations, the dominant ras oncogene and the p53 tumor suppressor gene, which are commonly found in lung cancer and pulmonary metastases. We discuss their specific roles in tumor development, invasion, metastasis formation, and their potential prognostic utility. In addition, we will discuss the concept of a novel modulator gene, ribonucleotide reductase, and review its role in the control of metastasis formation. PMID- 10368663 TI - Relationship between radical intensity and biological activity of cacao husk extracts. AB - The relationship between radical intensity and biological activity of cacao husk extracts was investigated. Electron spin resonance (ESR) spectroscopy demonstrated that the radical intensity of hexane, acetone, methanol and 70% methanol extracts increased with water-solubility. Several fractions of these husk extracts, separated by different column chromatographies, significantly inhibited the cytopathic effect of human immunodeficiency virus (HIV) infection in parallel with their radical intensity. However, their cytotoxic activity against human leukemic and carcinoma cell lines is not always correlated with their radical intensity. Water-soluble and lipophilic compounds might induce cytotoxic activity by different mechanisms. PMID- 10368662 TI - Drug control of solid tumour metastases: a critical view. AB - Metastasis of solid tumours represent the target of election for the pharmacological treatment of cancer. Nevertheless, commonly used treatments do not represent any selective approach, provided that drugs are mostly unspecific cytotoxics. Today many strategies adopted to interfere with metastasis growth concern the interaction with biological signals of the metastatic cells or of the host. One difference should be made between anti-metastatic and anti-metastasis drugs, in that only the latter realise the goal of selectively destroying metastasis wherever they are. In this context many agents active on newly identified molecular targets are more effective in preventing metastasis formation than in inhibiting their growth. NAMI-A, an innovative ruthenium compound, seems to provide optimism for the future and, in laboratory models, it is very active on lung metastases independently of the stage of their growth. The success of NAMI-A against metastasis should stimulate laboratory studies with appropriate experimental models to predict clinical activity, since the use of experimental conditions closely similar to those of human tumours should help the identification of more active compounds. PMID- 10368664 TI - Benzophenones as xanthine oxidase inhibitors. AB - Eight synthetic benzophenones were tested for their inhibitory effects on xanthine oxidase (XO). The enzyme, XO catalyses the oxidation of hypoxanthine to xanthine and of xanthine to uric acid, which has a lambda max of 295 nm, forming the basis for a spectrophotometric assay for the activity of XO. The results showed that 2,2',4,4'-tetrahydroxybenzophenone (6), 3,4,5,2',3',4' hexahydroxybenzophenone (8) and 4,4'-dihydroxybenzophenone (3) displayed the inhibitory effects on XO with an order of activity of IC50 = 47.59, 69.40 and 82.94 microM, respectively. The apparent inhibition constants (Ki) of (8) and (3) were 15.61 and 64.86 microM respectively, and both of them induced mixed type (non-competitive-uncompetitive) inhibitions of the substrate xanthine. PMID- 10368666 TI - Expression of Thrombomodulin in the epithelium of the urinary bladder: a possible source of urinary thrombomodulin. AB - Thrombomodulin (TM) is an endothelial thrombin receptor which acts as a natural anticoagulant through inactivation of the procoagulant activity of thromin. In the present study, we demonstrated that TM is expressed on the urinary bladder epithelium. The cell line BOY established from human transitional cell carcinoma of the urinary bladder also expressed the TM message (3.7kb). These cells express an 85 kDa TM protein which is identical in size to that of MEG-01, a human megakaryoblastic cell line, reported previously. We also ascertained the expression and localization of TM in the human urinary bladder by immunohistochemical staining. TM was localized in the membranes of the transitional epithelium and the cytoplasmic region of umbrella cells. The expression of TM in the transitional epithelium increased with worsening pathological status of cystitis. Based on these results, we concluded that TM is expressed in the urinary bladder. Further, soluble TM in urine may be partially derived from the urinary bladder. At present, the physiological significance of TM expression in the urinary bladder remains to be elucidated. PMID- 10368665 TI - Tumor infiltrating lymphocytes: CD8+ lymphocytes in canine transmissible venereal sarcomas at different stages of tumor growth. AB - This study utilized an in vivo tumor system (canine transmissible venereal sarcoma, CTVS) to assess wh correlation exists between tumor growth stage and phenotype of infiltrating lymphocytes. Additionally, the ability of CTVS cells to produce chemoattractants for canine lymphocytes was assessed. The CD8 subset of tumor infiltrating lymphocytes (TIL) during progressive growth, steady-state (no growth), and regression of the CTVS was determined. During progressive growth, the proportion of TIL expressing CD8 was significantly lower than that from regressing tumors (P < .001) and steady-state tumors (P < .01). Additionally, CTVS cell culture, CTVS spheroid cell culture, and CTVS spheroid/canine lymphocyte co-culture supernatants were tested for chemotactic activity for canine lymphocytes. CTVS and CTVS spheroid/canine lymphocyte co-culture supernatants both had significant chemotactic activity. Conversely, CTVS spheroid culture supernatants were negative for chemotactic activity for canine lymphocytes. These results indicate a correlation exists between the CD8 subset of TIL and clinical stage of the tumor. Also, we have shown that CTVS cells in vitro produce soluble factors that cause chemotaxis of canine lymphocytes. An understanding of the role of TIL and the ability of the tumor to attract them will be of help in designing strategies for immunomodulatory therapies for solid tumors. PMID- 10368667 TI - Effect of povidone-iodine on anastomotic tumor growth in an experimental model of colorectal cancer surgery. AB - Implantation of viable exfoliated tumor cells may be responsible for the local recurrence of colorectal cancer. Intraluminal colon cancer cells derived from chemically induced colon cancer in syngenic F344 rats, were introduced 2 cm proximally to a colonic anastomosis and anastomotic tumor growth was observed. Anastomotic tumor growth was found in about 30% of the animals, when sacrificed on the 3rd postoperative week. The cytotoxic efficacy of povidone-iodine (PVP-I) was tested with the same model. The administration of 10% PVP-I solution significantly reduced the incidence of tumor growth. Viable intraluminal tumor cells cause anastomotic tumor growth by becoming implanted on an anastomosis and PVP-I had effective cytotoxicity. PMID- 10368668 TI - Cimetidine enhances the antiproliferative effect of 5-fluorouracil on colon carcinoma SW620. AB - We investigated the effects of cimetidine on p-glycoprotein and on the antiproliferative effect of various anticancer drugs which are recognized by p glycoprotein. We used colon carcinoma SW620 cells and their doxorubicin resistant SW620 Ad300 derived cells, the latter express p-glycoprotein and have multidrug resistance phenotype. To assess the effect of cimetidine on the efflux activity of p-glycoprotein, we examined its effects on the accumulation of rhodamine123 which is recognized by p-glycoprotein. Cimetidine had no effect on rhodamine123 accumulation on either cell line. Cimetidine did not enhance the antiproliferative effect of any of the anticancer drugs investigated. This indicates that cimetidine was not recognized by p-glycoprotein. However, cimetidine remarkably enhanced the antiproliferation effects of 5-fluorouracil on SW620 cells, not those of 5-fluorouracil on SW620 Ad300, whereas cimetidine itself showed little effect on antiproliferation. This finding indicates that combination therapy using 5-fluorouracil and cimetidine might be useful for cancer patients. PMID- 10368669 TI - Interaction between sodium 5,6-benzylidene-L-ascorbate and gallic acid. AB - The interaction between sodium 5,6-benzylidene-L-ascorbate (SBA) and gallic acid was investigated by two different parameters: radical intensity and cytotoxicity induction. These compounds produced ESR signals of radicals under alkaline conditions. The addition of increasing concentrations of SBA completely scavenged the gallate radical and replaced the latter with its ascorbate radical. On the other hand, gallic acid dose-dependently enhanced the radical intensity of SBA. Both of these two compounds dose-dependently reduced the viable cell number of human squamous carcinoma HSC-2 cells without inducing internucleosomal DNA cleavage. Electron micrographs of the dying cells demonstrate the irreversible degenerative changes especially in the cytoplasm of the cells. The cytotoxic activity of gallic acid was almost completely eliminated by catalase, whereas SBA was totally insensitive to catalase. When these two compounds were mixed together before adding to HSC-2 cells, the cytotoxic activity of gallic acid was significantly reduced by SBA, whereas that of SBA was not reduced by gallic acid. SBA dose-dependently reduced the gallate oxidation in the culture medium. The interaction between SBA and gallic acid may modify their individual biological activity. PMID- 10368671 TI - Characterization of biological features and chemosensitivity of a new experimental lung metastasis model originating from the MXT mouse mammary adenocarcinoma. AB - The present study shows how an original mouse metastatic lung model was established from the MXT mammary adenocarcinoma. This metastatic model was obtained by injecting the C/MET clone into the tail veins of B6D2F1 mice. The C/MET clone corresponds to one of eleven cell clones that were isolated in vitro from the MXT model. Of these 11 clones, only the C/MET leads to lung metastatic tumor development when injected i.v. into mice. Furthermore, the C/MET clone colonizes the lung only. The present data show that the C/MET metastatic model and the MXT parental line are weakly (if reference is made to the P388 leukemia model) sensitive to adriamycin, clyclophosphamide and etoposide. However, under specific experimental conditions, the chemosensitivity of the C/MET model can be significantly increased. The C/MET model therefore appears to be an interesting pharmacological tool to test new investigational agents with anti-tumor potentialities to lung metastases. PMID- 10368670 TI - Antitumor protection using murine dendritic cells pulsed with acid-eluted peptides from in vivo grown tumors of different immunogenicities. AB - Peptides extracted from tumor cells after mild acid treatment can function as antigenic epitopes when presented by cultured dendritic cells. Peptides were extracted from four tumors syngeneic to C3H mice, three weakly immunogenic tumors (FSA, MCAK, HCA) and one non-immunogenic tumor (NFSA). Dendritic cells pulsed with peptides extracted from the three weakly immunogenic tumors partially protect mice from a tumor challenge with the parental cell line. This protection was evident by a slower rate of tumor appearance and a slower tumor growth curve when compared to control, non-immunized mice. However, vaccination of mice with dendritic cells pulsed with peptides derived from the non-immunogenic cell line NFSA did not elicit a protective response. Neither the route of immunization, the number of immunizations, nor the amount of peptides significantly affected the antitumor protection. Dendritic cells genetically engineered to produce IL-2 did not increase the protective effect of peptide-pulsed dendritic cells. These results suggest that only a partial protection against immunogenic tumors can be achieved when dendritic cells pulsed with acid-eluted tumor peptides are used as antitumor vaccination. PMID- 10368672 TI - Different H2 haplotypes have a strong influence on oncogene action. AB - The H2 complex has an important role in determining susceptibility to viral and chemical leukemogenesis in inbred mice. This also applies to transplantable leukemias, within the syngeneic system. In this respect H2K is sensitive, H2d is relatively sensitive, and H2b is absolutely resistant to leukemia induction and transplantation. In our present study we investigated the effect of Cyclophosphamide, (a known chemical leukemogen) on onco/suppressor gene expression in CBA/Ca mice, very shortly after treatment with chemical carcinogen without any manifestation of tumour/leukemia symptoms. Here we describe, in a "short-term" experiment, the gene expression of Ha-ras, c-myc and p53 which was similar to the leukemia induction in a "long-term" experiment. H2K showed marked elevation in terms of onco/suppressor gene expression. H2b expression was modest and H2d turned out to be more or less silent. The results obtained from a short term gene expression investigation shows similarity to those obtained earlier from long term leukemia inducing experiments. PMID- 10368673 TI - RT-PCR analysis of immune-modulating factors in PBMCs from patients with cancer: reduced IL-2 and increased IL-2-receptor (p55) expression characterize gastroenteric neoplasms. AB - BACKGROUND: To determine the potential contribution of cytokines associated with lymphocyte activation to the pathogenesis of immune impairment in gastroenteric cancer, we examined the expression of cytokine mRNA in peripheral blood mononuclear cells (PBMCs) and tumor-draining lymph nodes (LNs). MATERIAL AND METHODS: Using a reverse transcriptase-polymerase chain reaction (RT-PCR) technique, mRNA transcripts for interleukin (IL)-4, IL-2, IL-5, IL-6, IL-1 beta, IFN gamma, IL-10 and IL-2-receptor(IL-2R)(p55) were detected in PBMCs from 16 patients with gastroenteric cancer, undergoing surgical resection, and from 13 healthy donors. RESULTS: Patients expressed a different pattern of cytokines. Significantly increased IL-2R(p55) and reduced IL-2 mRNA expression were found in patients (p = 0.034, p = 0.043). IL-10 expression was significantly correlated to IL-2R(p55) expression in both groups (p = 0.011, p = 0.009). In LNs, the results reflected those from PBMCs of the same patient. CONCLUSIONS: Gastroenteric cancer showed a suppressive pattern of cytokine expression, suggesting an impairment of T cell activation that can be evidenced from the specific pattern of PBMC cytokine expression. PMID- 10368674 TI - How can one explain aneuploidy status in fibromatous and lipomatous naevus cell naevus? AB - 34 lightly fibromatous, 23 heavily fibromatous, 5 lipomatous and 10 naevus cell naevi were stained with Feulgen kit in order to evaluate their ploidy status with CAS 200 image analyzer. 26/34 lightly fibromatous, 18/23 heavily fibromatous, and 5/5 lipomatous naevi were either suspicious for aneuploidy (Auer III) or clearly aneuploid (Auer IV). In contrast all 10/10 naevus cell naevi were euploid. Proliferation (S-phase) was not increased in naevi fibromatously and lipomatously changed. The mechanisms leading to aneuploidy are discussed. PMID- 10368676 TI - Growth inhibition, nuclear uptake, and retention of anthracycline-formaldehyde conjugates in prostate cancer cells relative to clinical anthracyclines. AB - Recent data indicate that the clinical anthracycline anti-tumor drugs, doxorubicin (DOX), daunorubicin (DAU), and epidoxorubicin (EPI), catalyze the production of formaldehyde through induction of oxidative stress and bind the formaldehyde to form a metabolite which covalently bonds to DNA. Based upon this discovery, anthracycline-formaldehyde conjugates were synthesized and evaluated in three metastatic prostate cancer cell lines, LNCaP, PC-3, and DU-145. The doxorubicin-formaldehyde conjugate, Doxoform (DOXF), inhibits the growth of PC-3 and DU-145 cells 50- and 80-fold better, respectively, than the corresponding clinical drug, DOX. Daunorubicin- and epidoxorubicin-formaldehyde conjugates, Daunoform and Epidoxoform (DAUF and EPIF), inhibit the growth about 6 to 10-fold better than the clinical drugs, DAU and EPI. In addition, DAUF, DOXF, and EPIF are 2- to 20-fold more toxic to the doxorubicin-sensitive metastatic prostate cancer cell line, LNCaP. Fluorescence microscopy indicates that the nucleus is the major target for all six drugs. Flow cytometry together with fluorescence microscopy shows that DOXF and EPIF are taken up more rapidly and more abundantly and are retained in the nucleus longer than DOX and EPI, respectively, especially in DU-145 cells. The enhanced toxicity of the anthracycline-formaldehyde conjugates is attributed to their increased nuclear uptake and retention and suggests that DOXF, DAUF, and EPIF are prodrugs to the active metabolites of the clinical drugs DOX, DAU, and EPI. PMID- 10368675 TI - Immunohistochemical analysis of tumor antigen saturation following injection of monoclonal antibody G250. AB - Clinical tumor targeting studies with monoclonal antibody (mAb) G250 showed excellent targeting of primary renal cell carcinomas (RCC). However, tumor uptake decreased at higher mAb G250 doses, suggesting saturation of the G250 antigenic determinants. In this immunohistochemical study we investigated the saturability of G250 antigen sites after i.v. administration of mAb G250 at various protein doses in nude mice with RCC xenografts. Five groups of mice received five different protein doses (1, 3, 10, 30 or 100 micrograms) of murine mAb G250. Three days post injection mice were killed and the tumors were removed. Free G250 antigen sites, i.e., not targeted by the i.v. injected murine mAb G250 were determined by immunohistochemical staining with chimeric mAb G250. Distinct staining of the G250 antigen was observed only at the 1 and 3 micrograms dose whereas G250 antigen staining at higher doses was virtually negative. The results of this study indicate that saturation of antigen occurs at relatively low doses of i.v. administered mAb G250. Apparently, all antigenic determinants present on the RCC tumor cells were targeted, while previous preclinical studies suggested that i.v. administration of mAb G250 only saturated the accessible antigen sites. PMID- 10368678 TI - Topotecan-based combination chemotherapy for human malignant glioma. AB - BACKGROUND: Topotecan has been considered a promising agent for the adjuvant chemotherapy of human malignant glioma because of its novel mode of action, its activity against other solid tumors, and its good penetration across the blood brain barrier. However, the clinical effects of topotecan monotherapy in malignant glioma have been disappointing. MATERIALS AND METHODS: We sought to identify suitable partners for topotecan combination chemotherapy of malignant glioma using two well-characterized human malignant glioma cell lines, T98G and LN-229. The effects of co-exposure to topotecan and other chemotherapy drugs were assessed in cytotoxic and clonogenic cell death assays. RESULTS: We found additive, less-than-additive, or occasional antagonistic effects, but never synergistic activity of topotecan with either CCNU, VM26 or vincristine, in acute cytotoxicity or in clonogenic cell death assays, with simultaneous or sequential drug exposure. VM26 or vincristine followed by topotecan yielded the most favourable results. Further, prolonged exposure of the glioma cells to topotecan and either CCNU, VM26, vincristine, cisplatin, doxorubicin or cytarabine resulted in additive but not synergistic growth inhibition. CONCLUSIONS: The present study fails to identify a specific partner for topotecan-based combination chemotherapy of malignant glioma among the chemotherapeutic drugs examined here. While this does not exclude a possible synergy of the drug combinations examined here in vivo, a focus on novel partners for topotecan or topotecan-based chemoimmunotherapy may be more promising. PMID- 10368677 TI - Cytotoxic protein expression in non-Hodgkin's lymphomas and Hodgkin's disease. AB - Recent studies have shown that some peripheral T-cell lymphomas (PTCL) could be derived from lymphocytes with cytotoxic potential. Therefore, we have investigated by immunohistochemistry 34 cases of PTCL including 2 cases of hepatosplenic gamma delta PTCL and 5 cases of sinonasal NK-cell lymphomas as well as 7 cases of T-lymphoblastic lymphomas (T-LBL) for the expression of the cytotoxic proteins TIA-1 and granzyme B. In addition, 50 cases of Hodgkin's disease (HD) were investigated in order to see if these cytotoxic proteins are expressed by Hodgkin and Reed-Sternberg (HRS) cells. Expression of the TIA-1 is characteristic of cytotoxic cells regardless of their activation status whereas expression of granzymes is highly increased in activated cytotoxic cells. All the five cases of sinonasal NK-cell lymphomas expressed TIA-1 and granzyme B in most tumour cells. The two gamma delta PTCL cases expressed TIA-1 protein in most tumour cells but not granzyme B. Of the 32 other PTCL, 9 cases showed cytotoxic protein expression in tumour cells. These cases comprised 2 pleomorphic medium large cell (PML) (1 nodal and 1 intestinal) and 7 CD30 positive anaplastic large cell lymphomas (ALCL) (5 nodal and 2 cutaneous). Cytotoxic protein expression in our series appeared to be related to the location since 10/18 (55%) extranodal PTCL and NK-NHL and only 6/21 (28%) nodal PTCL expressed TIA-1, and related to histology since, in nodal PTCL, this pattern was observed in most anaplastic (5/6 cases) and in a few pleomorphic (1/9 cases) lymphomas, but not in AILD-type NHL (0/6 cases). The 7 cases of T-LBL did not express cytotoxic proteins in tumour cells. EBV was detected by EBER RNA in situ hybridization (RISH) in tumour cells in all 5 sinonasal NK-NHL and in scattered atypical cells in all 6 cases of AILD. Two of the 50 cases of HD weakly expressed TIA-1 and granzyme B in a proportion of HRS cells. EBV was detected by RISH in 19/50 cases of HD but no correlation was found between EBV status and expression of cytotoxic proteins in HRS cells. However, the finding that granzyme B positive cells were found very rarely in close vicinity of HRS cells suggests that the function of activated cytotoxic cells is locally inhibited by the HRS cells and/or the reactive cells in the vicinity of HRS cells. Taken together our data suggest that: a) sinonasal NK-cell NHL represent tumours of activated cytotoxic NK-cells, b) the hepatosplenic gamma delta PTCL represent tumours of nonactivated cytotoxic gamma delta T-cells, c) a small proportion of other PTCL, mostly anaplastic large cell lymphomas represent tumours of cytotoxic T-cells and d) only very few cases of HD expressing cytotoxic proteins in a proportion of tumour cells, could be derived from activated cytotoxic cells. PMID- 10368679 TI - Cell surface expression of urokinase receptor in normal mammary epithelial cells and breast cancer cell lines. AB - BACKGROUND: The urokinase receptor (uPAR) is important in the process of extracellular matrix degradation occurring during cancer cell invasion and metastasis. We wished to quantify uPAR on the surfaces of normal mammary epithelial cells (HMEC) and 6 well-known breast cancer cell lines using flow cytometry. MATERIALS AND METHODS: Cell surface uPAR was labelled with a monoclonal antibody, and this was detected with a fluorescent-labelled second antibody and accurately measured using flow cytometry. The measured fluorescent signals of the stained cells were interpolated with those of Quantum Simply Cellular bead standards to determine the number of uPAR sites per cell. RESULTS: The breast cancer cell lines ranged from 13,700 to 50,800 uPAR sites per cell, whilst HMEC cells had only 2,500 sites. CONCLUSIONS: This simple and reliable method showed that the expression of cell surface uPAR is higher in the breast cancer cell lines than in the normal mammary cells. PMID- 10368680 TI - Synthesis and cytotoxicity of aminosterols: activity studies on a non-small-cell bronchopulmonary carcinoma line (NSCLC-N6). AB - Various new aminosterols were synthesized. The antiproliferative activity of these compounds (I-IV) was studied in vitro on a continuous human non small-cell bronchopulmonary carcinoma line (NSCLC-N6) at the cell cycle level. The histograms indicate cell blockage in Phase Gl (compound I-III) associated with a reduction in the number of cells phases S and G2M and appearance of cellular debris derived from cells in Phase G1. PMID- 10368681 TI - Endonuclease activity and hydrogen peroxide-induced cytotoxicity in human glioblastoma and glioma cell lines. AB - Hydrogen peroxide (H2O2) induced internucleosomal DNA cleavage in human myelogenous leukemic cell lines (HL-60, ML-1, THP-1, U-937), but not in human glioblastoma (T98G, U87MG) and glioma (KG1C) cell lines. However, H2O2 produced apoptotic cells, characterized by cell shrinkage, nuclear fragmentation and chromatin condensation in glioblastoma and glioma cell lines. Autodigestion experiments revealed that the major endonucleases, present in all leukemic, glioblastoma and glioma cell lines, were divalent cation-independent endonuclease(s). The endonudease(s) present in the lysates of all these cells were activated at acidic, but not at neutral pH. The results suggest that the endonuclease activity might be differently regulated between leukemic and glioma cell lines. PMID- 10368682 TI - Use of CDC2 from etoposide-treated cells as substrate to assay CDC25 phosphatase activity. AB - Cyclin-dependent kinases (CDKs) regulate the key transition of the cell cycle in all organisms. In response to Etoposide (VP-16) induced DNA damage, cells undergo a G2-phase arrest resulting in the accumulation of inactive CDK1 (CDC2) kinase complexes. Here we report that upon Etoposide treatment CDC2 is phosphorylated on tyrosine 15 and is dephosphorylated and activated in vitro by recombinant CDC25 phosphatase. We also show that inactive CDC2 kinase from Etoposide-treated cells can be used as a substrate in a sensitive two-step assay of CDC25 phosphatase. This assay, which is very simple to set-up, is based on the monitoring of CDC2 kinase activity after CDC25-dependent dephosphorylation. It provides the possibility to use a highly physiological substrate in antimitotic drugs screening. PMID- 10368683 TI - Falsification of tetrazolium dye (MTT) based cytotoxicity assay results due to mycoplasma contamination of cell cultures. AB - Mycoplasma contamination of cell cultures is a frequently observed problem. Due to the inconspicuous growth in cell cultures, periodical screening procedures represent the only protection. Many influences of mycoplasma on cell culture parameters have been described. We addressed the question of whether mycoplasma contamination affects the most frequently used cytotoxicity assay, the tetrazolium based MTT assay. We contaminated C6 glioma cells with mycoplasma and performed MTT assays with doxorubicin, vincristine, etoposide and cisplatinum under various conditions. Contaminated cells demonstrated significant different results when tested with the MTT assay than mycoplasma free controls. Differences were not detectable when cells were counted as toxicity assay. Due to an additional reduction of tetrazolium by mycoplasmas, contaminated cells appeared up to 15 fold resistant to doxorubicin, vincristine and etoposide, but not to cisplatinum. Differences decreased with decreasing drug doses and decreasing plated cell count. Our findings confirm the compelling need for periodical mycoplasma screening, especially when tetrazolium based cytotoxicity assay (MTT) are used. PMID- 10368684 TI - Anticancer potential of N,N-diethylaminoethyl ethers of flavanone oximes: a comparison with mitoxantrone action on rat Yoshida sarcoma cells in vivo. AB - This study was performed to evaluate the anticancer abilities of four biologically active N,N-diethylaminoethyl ethers of flavanone oximes against rat Yoshida Sarcoma cells in vivo, and to investigate the mechanism(s) involved. The effects were compared with those of anthraquinone drug (mitoxantrone) action. The presented results provide the first evidence that all the investigated substances induce programmed cell death (apoptosis) of Yoshida Sarcoma cells in vivo. On interpretative grounds, the administration of investigated flavanone derivatives in the promotion phase of the disease led to both growth inhibition (cell cycle perturbation) and apoptosis. A correlation was found between structure of the substituent(s) at B-ring of substances and the revealed anticancer activity. The data suggest that flavanone derivatives (oxime ethers) besides their antiradical, antioxidant and radioprotector properties observed before, may act as promising anticancer agents acting in the promotion phase of disease. This finding prompted us to consider the development of a new strategy: modulation of effects using combination therapy involving mitoxantrone and flavanone oximes. PMID- 10368685 TI - Antiproliferative and cytotoxic effects of N,N-diethylaminoethyl ethers of flavanone oximes: a comparison with anthracycline(s), anthraquinone and nitroxides action. AB - Since flavanone oximes derivatives (ethers) have been shown to modulate the growth of Yoshida Sarcoma cells in vivo and to induce apoptosis, the effects of these substances on immortalized cell lines growth were examined. Cell viability and sensitivity to investigated substances was measured by the modified tetrazolium salt (MTT) assay. The antiproliferative effects were expressed as IC50 and IC90, respectively. There were very substantial differences in the dose dependency of the observed antiproliferative and cytotoxic effects. The structure activity relationship was evident and revealed that the substitution at B-ring of molecule seems to be an important factor in flavanone oxime (ether) potency. Compared to anticancer drugs (doxorubicin, aclarubicin and mitoxantrone) flavanone oximes displayed cytotoxicity at considerably higher concentrations. The antiproliferative action of the investigated model nitroxides depended on the free radical part of the molecule. N-hydroxy derivative (reduced cation form) did not influence cells proliferation and nor display any cytotoxicity at the applied range about 60 times higher than those of flavanone derivatives. Taken together it seems reasonable to suggest that flavanone oxime(s) (ethers) as compared with antracycline(s), anthraquinone and nitroxides might be especially good candidates for in the future development of new therapeutic techniques. PMID- 10368688 TI - Comparison of annamycin to adriamycin in cardiac and MDR tumor cell systems. AB - Based on the response of a wide variety of tumors to the anthracycline, Adriamycin, numerous studies have been initiated to find an even more effective analog. In this pursuit two of the obstacles that have been necessary to overcome are a unique dose dependent Adriamycin-induced cardiotoxicity reported in patients treated with this chemotherapeutic agent as well as p-gp-mediated multi drug resistance (MDR) which has been found in tumor cells exposed to Adriamycin in vitro and in vivo as well as in human tumor samples. Using an in vitro cardiac cell system and MDR+ and MDR- Friend leukemia cell lines we find that a relatively new anthracycline, Annamycin, has reduced cardiotoxic activity but is more effective in inhibiting the growth of MDR+ cells than Adriamycin. The reduced cardiotoxicity of Annamycin is approximately 10 fold lower than Adriamycin whereas the increased efficacy against the MDR+ Friend leukemia tumor cell line is about 2 fold. The observation that Adriamycin preferentially accumulates in cardiac-muscle (CM) but not in cardiac non-muscle (NM) cells while Annamycin accumulates equally in both, may explain in part the reduced cardiotoxicity of Annamycin. Moreover, the cytosolic accumulation of Annamycin vs the nuclear localization of Adriamycin suggests a different target site for each drug. PMID- 10368687 TI - Microsatellite instability is co-selectable with gene amplification in a mammalian mutator phenotype. AB - The effect of an unbalanced nucleotide pool on the stability of dinucleotide (CA)n microsatellite sequences was investigated in the mutator phenotype CSA7 clone isolated from Chinese hamster CHEF18 cell line. A series of clones isolated from CHEF18 and CSA7 cells and resistant to 6-thioguanine (TG) were shown to be stable at the three examined microsatellite loci. Furthermore, the clones isolated from CHEF18 cells and resistant to N-phosphonacetyl-L-aspartate (PALA) were stable, whereas those isolated from CSA7 clone were unstable. At the biological level the clones with instability did not show tolerance of methylnitrosourea-induced damage, thus excluding the presence of a defective mismatch repair. On the contrary, the molecular characterization showed that the instability, measured as extension or contraction of (CA)n repeats, involved numerous repeats resembling the amplification rather than mutation of the microsatellite loci. The results therefore indicate that the unbalanced nucleotide pool of CSA7 clone influences the overall rate of gene amplification and support that the unstable microsatellites are coselectable with other gene amplification events. PMID- 10368686 TI - Effect of citrus flavonoids on HL-60 cell differentiation. AB - Twenty-seven Citrus flavonoids were examined for their activity of induction of terminal differentiation of human promyelocytic leukemia cells (HL-60) by nitro blue tetrazolium (NBT) reducing, nonspecific esterase, specific esterase, and phagocytic activities. 10 flavonoids were judged to be active (percentage of NBT reducing cells was more than 40% at a concentration of 40 microM), and the rank order of potency was natsudaidain, luteolin, tangeretin, quercetin, apigenin, 3, 3, '4, '5, 6, 7, 8-heptamethoxyflavone, nobiletin, acacetin, eriodictyol, and taxifolin. These flavonoids exerted their activity in a dose-dependent manner. HL 60 cells treated with these flavonoids differentiated into mature monocyte/macrophage. The structure-activity relationship established from comparison between flavones and flavanones revealed that ortho-catechol moiety in ring B and C2-C3 double bond had an important role for induction of differentiation of HL-60. In polymethoxylated flavones, hydroxyl group at C3 and methoxyl group at C8 enhanced the differentiation-inducing activity. PMID- 10368689 TI - 5,6 Dihydro-5'-azacytidine (DHAC) restores androgen responsiveness in androgen insensitive prostate cancer cells. AB - INTRODUCTION: The androgen resistance of some prostate cancer patients may be due to transcriptional inactivation of the androgen receptor (AR) gene catalyzed by cytosine DNA methyltransferase. MATERIALS AND METHODS: To determine if an inhibitor of cytosine DNA methyltransferase, 5,6-dihydro-5'-azacytidine (DHAC), can restore the androgen sensitivity in androgen-insensitive human prostate carcinoma cell lines in vitro, we cultured androgen-insensitive (PC3, DU-145, and TSUPrl) and androgen-responsive (LNCaP) cells with subcytotoxic concentrations (< or = IC50) of DHAC for 14 days followed by exposure to dihydrotestosterone (DHT) or to hydroxyflutamide for 7 days. RESULTS AND CONCLUSIONS: Only DHAC-treated DU 145 cells showed growth stimulation by 10(-11) to 10(-9) M DHT and a partial inhibition by 10(-5) and 10(-6) M hydroxyflutamide. However, since DU-145 is the only cell line tested that is known to have a hypermethylated AR promoter, the observed effects may be due to a partial demethylation of the AR by DHAC. Our data provide an evidence that cytosine DNA methyltransferase inhibitors can restore androgen responsiveness in androgen-refractory tumor cells, which are then sensitive to growth inhibition by antiandrogens. PMID- 10368691 TI - Cytotoxic effects of MGI 114 are independent of tumor p53 or p21 expression. AB - MGI 114, an analog of illudin S, shows potent activity against a broad range of human tumors in vitro and in vivo, including drug resistant tumors. In this study we examined cytotoxicity of MGI 114 against human tumor cell lines (MCF7, MDA.MB.468, EJ1, J82, SCaBER, KG-1, HL60, and IMR-90) with differing expression of p53 and/or p21 (WAF1) tumor suppressor genes. Only MCF7 and IMR-90 express the wild type p53, WAF1 is present in high levels in MCF7 and SCaBER. WAF1 expression can be induced in KG-1, HL60, and IMR-90. The cells were treated with MGI 114 at 0.1, 1.0 and 10 micrograms/ml in 1 h exposure and with 0.01, 0.1 and 1.0 microgram/ml MGI 114 in a continuous exposure. Cell numbers were measured at days 2, 4, and 7. MGI 114 suppressed growth in all cell lines at day 2 after 1 h exposure at the two highest concentrations and at all concentrations in a continuous exposure. Some cells partly recovered from the inhibition by day 4. Expression of WAF1 had no apparent effect on growth suppression by MGI 114, however, cells with inducible WAF1 showed slower recovery from MGI 114 inhibition in comparison with the cells under non-permissive conditions. Overall, MGI 114 effectively inhibited growth of human cancer cells regardless of their p53 and WAF1 status. PMID- 10368692 TI - A PCR-aided transcript titration assay (PATTY) to measure topoisomerase I gene expression in human tumor specimens. AB - Topoisomerase I (topo I) inhibitors are promising anticancer agents with demonstrated activity against a wide range of solid tumors. Quantitative information on topol mRNA levels in tumor biopsies may predict response to topo I inhibitors. MATERIALS AND METHODS: A polymerase chain reaction aided transcript titration assay (PATTY) was developed to allow quantitation of topol mRNA in small samples. Concentrations of topol mRNA in total RNA samples were estimated by RT-PCR analysis in a human small cell lung cancer (SCLC) cell line (GLC,) and its topotecan (GL2C/SK and F) and camptothecin (GL2C/Campt) resistant sublines, human non-small cell lung cancer (NSCLC) and ovarian carcinoma samples. RESULTS: Topol PATTY showed a decreased topo I mRNA level in GLC2/SK and F (4.5 pg/100 ng total RNA) and GLC,/Campt (2.2 pg/100 ng total RNA), respectively, compared to the parent cell line GLC2 (5.4 pg/100 ng total RNA). Topol protein levels as measured by Western blotting were compatible with topol mRNA levels. Median (range) topol mRNA levels were 3.23 (2.33-5.10) pg/100 ng total RNA in resected NSCLC specimen (n = 6), and 2.03 (0.54-0.95) pg/100 ng total RNA in resected ovarian cancer specimen (n = 6). CONCLUSION: We conclude that topol PATTY is a new assay that quantitates topol mRNA levels in cell lines and small tumor samples. PMID- 10368690 TI - 5,6-Dihydro-5'-azacytidine (DHAC) affects estrogen sensitivity in estrogen refractory human breast carcinoma cell lines. AB - BACKGROUND: There is little effective therapy for patients with hormone refractory breast cancer. Hormone resistance is frequently due to the transcriptional inactivation of the estrogen receptor (ER) gene. We determined the effect of DHAC, a cytosine DNA methyltransferase (CMT) inhibitor, on the estrogen sensitivity in three human breast carcinoma cell lines with intermediate to low levels of estrogen receptor (ER) expression: MCF7 (adriamycin-sensitive), MCF7M/Adr (adriamycin-resistant), and MDA-435, and one ER+ cell line, ZR75-1. MATERIALS AND METHODS: Cells maintained in culture were exposed to DHAC or vehicle continuously for 14 days, then exposed to estradiol or tamoxifen and counted on day 21. RESULTS: Exposure to DHAC did not affect estrogen sensitivity in ZR-75-1 and MCF7M/Adr cells. DHAC treatment of MCF7 and MDA-435 cells resulted in significant (p < 0.05) growth stimulation in response to estrogen at 10(-6) M, and to growth modulation by tamoxifen at 10(-5) to 10(-7) M. CONCLUSIONS: These data suggest that DHAC can restore the estrogen sensitivity in ER-breast cancer. Thus, DHAC and other novel CMT inhibitors may have a clinical application in treating estrogen-refractory breast cancer patients by restoring the estrogen sensitivity and allowing these patients to respond again to conventional therapy with estrogen antagonists. PMID- 10368693 TI - Toxoplasma infection and cell free extract of the parasites are able to reverse multidrug resistance of mouse lymphoma and human gastric cancer cells in vitro. AB - A large number of compounds are known to reduce the ATP-dependent efflux pump activity of multidrug resistant (mdr) tumor cells. Here we report that an infection of cancer cells with T. gondii reduced the multidrug resistance of the tumour cells against cytostatic drugs. Two mouse lymphoma cell lines (Mdr L 5718 and Par 5718) were infected with Toxoplasma gondii in vitro and the reduction of efflux pump activity of the cells was measured. The drug accumulation (Rhodamin 123) was increased in the infected mdr cell lines compared with non- infected mdr cells, and no effect was shown after infection of the parental cell line. The same effect was also achieved by incubation of Mdr-tumor cells with cell lysate of Toxoplasma gondii. Mdr-1-gene expression was reduced in the infected cell lines 48 hours after infection. Co-cultivation of Toxoplasma gondii with mdr cell lines separated by a microfilter from tumor cells was performed, but this cocultivation did not change the mdr efflux activity. The effect of Toxoplasma gondii infection on the efflux pump activity and mdr-1 gene expression was also examined in the human gastric cancer cells. A sensitization of resistant gastric cancer cells was also achieved by parasite infection. This phenomenon is an evidence that a reduction of resistance in tumor cells can be achieved by a natural parasite infection. It is as yet unclear whether an active infection or another substance of T. gondii is responsible for this phenomenon. PMID- 10368694 TI - Loss of heterozygosity of TP53 is not correlated with clinicopathological variables in sporadic colorectal carcinomas. AB - We have analyzed the loss of heterozygosity (LOH) of TP53 in a series of 96 sporadic colorectal carcinomas by means of PCR, using two microsatellite sequences (TP53 and Mfd152), to investigate its possible relationship with several clinicopathological variables in the Spanish population. Forty six of the 96 patients (48%) showed loss of one allele of the microsatellite TP53, Mfd152 or both, when compared with normal colorectal mucosae and blood samples of the same patient. This high percentage of LOH seems to corroborate the important role of p53 in sporadic colorectal cancer. However, we have found that LOH on this region is independent of histological grade and tumour location. With regard to tumour Dukes' stage, the fact that a substantial proportion of tumours show LOH on 17p from the first stages of the disease could imply that this alteration is not related with the invasiveness acquisition staging. PMID- 10368695 TI - Clinical analysis of the novel breast cancer serum assay BT1. AB - In an effort to define possible clinical relationships to the BT1 serum assay, a retrospective study population including both Caucasian and African American breast cancer patients was tested. Test results were compared to clinical information provided by the Virginia Cancer Registry. The 64 patients, ages 29 through 69, had a wide range of BT1 values which were not attributable to race or age. The majority of these patients had infiltrating duct carcinoma, with eight additional morphology of neoplasm diagnoses represented in the entire group. No morphology code was associated with either a reactive or non-reactive BT1 result. Staging information was available for 26 patients, diagnosed with stages I, II, and III breast cancer. Positive BT1 values were found throughout these diagnosis stages. In addition, multiple serum samples were available from some patients. Analysis of these longitudinal samples showed different patterns, with test values remaining unreactive in 4 patients, and reactive in 9 others. Interestingly, 5 patients showed values increasing from negative to positive while 3 patients went from positive to negative over time. PMID- 10368696 TI - Dietary supplementation of selenomethionine reduces metastasis of melanoma cells in mice. AB - The present study investigated the effect of dietary supplementation of selenomethionine on pulmonary metastasis of B16BL6 murine melanoma cells in C57BL/6 mice. Mice were assigned to four groups of 15 each. They were fed a basal AIN93G diet and the basal diet supplemented with 2.5 ppm or 5 ppm selenium as selenomethionine or with 2.5 ppm selenium as selenite for two weeks before and after the intravenous injection of 0.5 x 10(5) tumor cells. At necropsy, the number and size of tumors that developed in the lungs were determined. The number of mice that had > or = 11 tumors was 13, 8, 8, and 6 (p < 0.02 compared with the control), and the median number of lung tumors was 64, 14, 12 (p < 0.05 compared with the control), and 8 (p < 0.01 compared with the control) in the control group and the groups with 2.5 ppm and 5 ppm selenium as selenomethionine and 2.5 ppm selenium as selenite. Dietary supplementation of selenomethionine decreased tumor cross-sectional area and tumor volume compared with the controls. At the same dietary level, selenite had a greater inhibitory effect on tumor size than selenomethionine. These results demonstrate that dietary supplementation of selenomethionine reduced experimental metastasis of melanoma cells in mice and inhibited the growth of metastatic tumors that formed in the lungs. It is concluded that selenomethionine is an active form of selenium that reduces experimental metastasis. PMID- 10368697 TI - Levalbuterol for asthma. PMID- 10368698 TI - Interferon plus ribavirin for chronic hepatitis C. PMID- 10368699 TI - Generic drugs. PMID- 10368700 TI - Miglitol for type 2 diabetes mellitus. PMID- 10368701 TI - A vaccine for rotavirus. PMID- 10368702 TI - Sunscreens: are they safe and effective? PMID- 10368703 TI - Cilostazol for intermittent claudication. PMID- 10368704 TI - Advice for travelers. PMID- 10368705 TI - [Orthopnea: not always of cardiac origin]. AB - Respiratory insufficiency developed in a man aged 68 after cardiac surgery and in a man aged 60 with COPD and a history of cigarette smoking after an attack of 'flu', while in a woman aged 70 with non insulin-dependent diabetes mellitus it had been present for years. All three had bilateral diaphragmatic paralysis. The diagnosis is based on the triad orthopnoea, paradoxical abdominal movements during respiration in the recumbent position and a decrease of the vital capacity in the horizontal as compared with the sitting position. The patients' physical condition could be improved with the aid of (noninvasive) ventilatory support. PMID- 10368706 TI - [Analysis and treatment of pancreatic cystic diseases]. AB - Owing to the spectacular progress in imaging techniques, cystic lesions of the pancreas are detected more often than previously, and this leads to therapeutic dilemmas. Of the cystic lesions of the pancreas, 80% are found to be pseudocysts and 10-15%, neoplastic cysts. The definition of a pseudocyst is: 'an accumulation of pancreatic juice surrounded by a wall of connective tissue or granulation tissue, developed as the result of acute pancreatitis, pancreatic injury or chronic pancreatitis'. In cases of asymptomatic pseudocyst, an expectative policy suffices; growth and symptomatic pseudocysts justify intervention. In addition to surgical internal drainage (cystojejunostomy) there are new therapeutic techniques: external drainage guided by ultrasonography or CT, and internal drainage guided by endoscopy. Endoscopic drainage is the treatment of choice, but it requires experienced hands. The cystic tumours are subdivided into two groups: serous and mucinous cystadenomas. The group of mucinous tumours is subdivided into mucinous cystadenomas and intraductal papillary mucinous tumours. The mucinous or macrocystic adenoma is potentially malignant and should be treated as a malignancy. PMID- 10368707 TI - [Ultraviolet A-I phototherapy for skin diseases]. AB - Favourable effects of sunlight on various skin diseases include inhibition of rapid proliferation of cells (psoriasis), modulation of cells in an inflammatory infiltrate (atopic eczema) and stimulation of proteolytic enzymes (scleroderma). The ultraviolet (UV) fraction of the solar spectrum is the most biologically active because it is almost completely absorbed by the skin. UVB and the combination of psoralens with UVA (PUVA) have become important therapeutic modalities, especially for psoriasis and eczema. Lamps producing long wave UV radiation are available: UVA-I light. Owing to its longer wavelength it penetrates more deeply into the skin and gives less risk of development of skin cancer than other forms of UV radiation. Good results are reported of application of UVA-I in patients suffering from atopic dermatitis, scleroderma, urticaria pigmentosa, and systemic lupus erythematosus. PMID- 10368709 TI - [Practice guidelines for diagnosis of vision disorders in mentally handicapped persons. National Organization for Quality Assurance in Hospitals]. AB - Visual impairment is more frequent among people with intellectual disability than among the general population. Because the diagnosis often fails to be made in this group screening is justified. Timely recognition may have consequences for prevention and treatment. Therefore, a consensus statement has been developed by the Dutch associations of physicians for intellectual disability, general practitioners, ophthalmologists, paediatricians, youth health physicians and orthoptists. Specialized ophthalmological assessment of all young children with retarded development is recommended, as well as assessment of visual function at the ages of 3, 6, 12 and 18 years and every 5 years from the age of 45. In adults with Down's syndrome, an additional assessment at the age of 30 is recommended for detection of age-related cataract and (increased) refractive errors. Most people can be assessed by the general practitioner, using normal methods. Diagnostic methods applicable in insufficiently co-operative people have been recommended. Low-threshold regional expert centres for diagnosis in children and adults who are difficult to assess and for specialized guidance would be advisable. PMID- 10368708 TI - [Two new antidepressants: mirtazapine and venlafaxine]. AB - Mirtazapine and venlafaxine are two novel antidepressants which are pharmacologically different from other new anti-depressants. Mirtazapine blocks the presynaptic alpha 2-adrenoreceptors and has a benign safety profile. Venlafaxine particularly inhibits the reuptake of serotonin and in higher doses it inhibits the reuptake of norepinephrine as well. Its adverse events profile is similar to that of the selective serotonin reuptake inhibitors (SSRIs). Efficacy studies show that both compounds are effective in depressed outpatients. In depressed inpatients with psychotic features, suicidality or a longlasting episode mirtazapine is not effective and in such patients venlafaxine was not studied. PMID- 10368710 TI - [Laparoscopic donor nephrectomy for kidney transplants from living family members: good preliminary results]. AB - OBJECTIVE: Evaluation of safety and technical feasibility of laparoscopic live donor nephrectomy. DESIGN: Descriptive. METHOD: The per- and postoperative results were analysed of 15 patients subjected to laparoscopic live donor nephrectomy in the Erasmus Medical Centre Rotterdam, Dept. of General Surgery, the Netherlands. Both left and right nephrectomy were performed via the transperitoneal route. The kidney was removed via a subumbilical incision. RESULTS: Laparoscopic donor nephrectomy was attempted in 15 patients and completed successfully in 14. Conversion to flank incision was resorted to one patient because of a venous bleeding. Median operating time was 290 min (SD: 57). Mean warm ischaemia time was 7 min (range: 4-17), including laparoscopic harvest. All kidneys were functioning well after transplantation. The mean duration of postoperative hospitalization of the donors was 4 days. CONCLUSION: Laparoscopic live donor nephrectomy is a safe and technically feasible procedure in a kidney transplant programme involving a living relative. PMID- 10368711 TI - [Total prostatectomy--a good option for residual prostate carcinoma after I-125 implantation of external radiotherapy]. AB - OBJECTIVE: To evaluate the results of salvage prostatectomy after previous radiation therapy for locally confined prostate cancer. DESIGN: Retrospective. METHOD: Data were collected from the records of all patients with prostate cancer who underwent salvage prostatectomy after I-125 implantation or external radiation therapy in the Netherlands Cancer Institute/Antoni van Leeuwenhoek Hospital, Amsterdam, the Netherlands, 1991-1997. Indications for surgery were: locally confined histologically proven residual cancer, good life expectancy, fit for surgery. Standard preoperative workup was done together with a tumour marker measurement, transrectal ultrasound with biopsy of the prostate and a bonescan. Per- an postoperative complications, pathology result and postoperative PSA were assessed. Progression free survival, overall survival and cancer specific survival were calculated according to the Kaplan-Meier method. RESULTS: 10 patients with a mean age of 67.2 years (range: 57-79) and a median follow up of 78 months (range: 0-89) underwent a total prostatectomy after I-125 implantation (7 patients) or external radiation therapy (3 patients). One patient died after the operation from acute tubular necrosis. One patient developed an internal hernia, requiring surgery. Four patients needed pads during the daytime for stress incontinence for urine. The 5-year progression free survival was 72% (95% confidence interval (95% CI): 44-100), the overall survival was 90% (95% CI: 73 100) and the cancer specific survival was 90% (95% CI: 73-100). No local recurrences were detected. CONCLUSION: The local control and the 5-year survival were good in this selected patient group. PMID- 10368712 TI - [Myositis in the presence of slightly elevated creatine kinase values]. AB - The clinical picture with, among other things, muscular swelling, fever and nocturnal sweating in three males aged 30, 37 and 52 years, suggested a neoplasm and sepsis. Ultimately, they were found to suffer from focal myositis, localized nodular myositis and polymyositis, respectively. The ESR and leukocyte counts were increased, the serum creatinine kinase (CK) activity was normal or slightly increased. The symptoms decreased after prednisone treatment. Both localized and generalized inflammatory muscular diseases may be present without the serum CK activity being raised. PMID- 10368713 TI - [Clinical thinking and decision making in practice. A young man in shock]. PMID- 10368714 TI - [Clinical thinking and decision making in practice. A young man in shock]. PMID- 10368715 TI - [A strontium-89 injection: a simple treatment of painful bone metastases in patients with prostate cancer unresponsive to hormonal treatment]. AB - Four patients (men aged 75, 67, 65 and 69 years) with painful osseous metastases from prostate cancer were treated by intravenous radionuclide therapy using Strontium-89. All had secondary progression after initially successful hormonal treatment. Three of these four had good responses lasting from 5 to 9 months. One patient with rapidly progressive disease did not respond. Second and third injections were successful in two patients. Mild bone marrow suppression was observed in all, but was not clinically significant. The 70-80% chance of long lasting pain alleviation through a single injection of Strontium-89 is a valuable addition in the treatment of painful bone metastases from prostate cancer, and probably also in such metastases from breast cancer. PMID- 10368716 TI - [Immunology in medical practice. XIX. Etiology and pathogenesis of auto-immune diseases]. AB - In spite of the collection in the last 50 years of an enormous amount of knowledge about aetiology and pathogenesis of autoimmune diseases, important questions have not yet been answered (adequately), as shown by the very limited therapeutic application of this knowledge. Although autoreactive B and T cells are demonstrable in healthy individuals, this does under normal circumstances not lead to disease, for instance because necessary co-factors are lacking, B lymphocytes for antibody production require help from suitable autoreactive T helper cells, because of the action of T suppressor cells and due to anti idiotypic antibodies and T cell receptors that may be directed against the antigen receptors of autoreactive B and T cells. Autoimmune diseases are sometimes subdivided into organspecific (e.g. Graves' disease) and systemic autoimmune diseases (e.g. disseminated lupus erythematosus). Roughly speaking there are two hypotheses concerning the aetiology or development of autoimmune diseases. The first hypothesis assumes a specific antigen as the driving factor, the second dysregulation of the immune system. The new insights into the pathogenesis of autoimmune diseases so far led to development of only few new drugs or therapies. PMID- 10368718 TI - [New WHO-classification of lung and pleural tumors]. AB - A new classification of the World Health Organization (WHO) of lung and pleural tumours will be published presently. Compared with the previous edition of 1981 the changed parts more accurately reflect the available therapeutic choices and the prognostic characteristics of the different tumour types. The classification is based on conventional light-microscopical typing. Additional techniques (from histochemistry, immune histochemistry, electron microscopy and molecular biology) have not yet decisive influence on tumour typing. The dichotomy between small cell and large-cell carcinomas is too simplistic, as the group of large-cell carcinomas is heterogeneous, and further differentiation leads to identification of tumour types with distinct therapeutic options and prognostic characteristics. There are new criteria for the classification of neuroendocrine tumours, such as the mitotic index. It is recommended to use the newly revised classification for diagnostic purposes, epidemiology and biologic studies. PMID- 10368717 TI - [Immunology in medical practice. XX. Organ specific autoimmune diseases]. AB - Organ specific autoimmune diseases are predominantly diseases of the endocrine glands and involve amongst others the thyroid (Hashimoto's disease, primary myxoedema, Graves' disease), the islets of Langerhans (type I diabetes mellitus) and the adrenals (Addison's disease). Over the past fifty years the knowledge on the pathogenesis of these diseases has considerably increased, leading to a large number of newly developed diagnostic tools, particularly determination of autoantibodies. Most of these autoimmune diseases have a (long) subclinical latency period. During this latency period a reliable prediction of later clinical manifestation is feasible. Preventive interventions during the latency period to correct underlying abnormalities of the target endocrine gland and/or the immune system are currently being tested in experimental animal models. PMID- 10368719 TI - [Drug induced alopecia]. AB - Hairs are produced in hair follicles that have a cyclic activity. The cycles that normal hair follicles go through are the metabolically active anagen phase, the catagen transitional phase and the resting telogen phase. Loss of more than 120 hairs daily is called alopecia. Drugs typically cause a diffuse, reversible alopecia by influencing one of the cycles that hair follicles go through. Because hair loss can have many causes, a causal relationship between a suspected drug and hair loss may be hard to prove. The principal categories of drugs that may cause alopecia listed in the literature are cytostatics, anticoagulants, interferons, tretinoid derivatives and lithium carbonate. The Netherlands Pharmacovigilance Foundation LAREB has received reports of alopecia as side effect of antimalarials, beta-receptor blocking agents, sex hormones, ACE inhibitors, angiotensin II antagonists and anticoagulants. PMID- 10368720 TI - [Prevention of a suspected epidemic of methicillin-resistant staphylococcus aureus (MRSA) in a nursing home]. AB - EPIDEMIC: Following the identification of an index patient with meticillin resistant Staphylococcus aureus (MRSA) in a Dutch nursing home with 175 residents in the south of Limburg province, the Netherlands (microbiological diagnosis obtained from a foreign laboratory), a survey was carried out to trace contacts by means of the 'ring principle' of outbreak management. If positive cultures were found in the first ring of residents the contact and source tracing was extended. According to the Dutch guidelines for MRSA in nursing homes many preventive measures were taken regarding colonised residents and employees and the cleaning of rooms. Ten days after the occurrence of the index, 29 persons, 9 employees and 20 residents, were diagnosed as colonised with MRSA. Because of this extraordinary count compared with earlier Dutch findings (only 0.16% of inhabitants colonised) there were doubts about the laboratory results. A counter expertise from a Dutch lab and the National Institute of Health and Environmental Hygiene showed no MRSA, but meticillin-sensitive S. aureus. DISCUSSION: This alleged epidemic had very aggravating consequences for residents and employees and large financial consequences for the nursing home. There was a good reaction to the crisis by a multidisciplinary team with external specialists. The Inspectorate of Health emphasized the importance of standardized quality and interpretation of laboratory results by microbiological experts. This should be kept in mind when contracting foreign laboratories specially because the Dutch policy is to strictly avoid MRSA in intramural setting. Verification of diagnosis proved again to be an essential step in outbreak management. PMID- 10368721 TI - [Follow-up after melanoma resection is more extensive than recommended in the practice guidelines, primarily for reassurance of the patient]. AB - OBJECTIVE: To determine to what extent the follow-up after resection of melanoma in practice corresponds to the relevant guidelines in the first revised version of the consensus 'Melanoma of the skin'. DESIGN: Descriptive, retrospective. METHOD: For 67 patients, who had a melanoma resected in September 1993-April 1995 in the Academic Hospital, Vrije Universiteit, Amsterdam, the Netherlands, data were collected in May-August 1997 on the first two years of follow-up from the medical records (n = 42) and through communication in writing with the referring physicians and the physicians involved in the follow-up in other hospitals in the Netherlands (n = 25). The frequency of physical examination and routine diagnostics by the doctor was evaluated. To gain insight into the reasons why in some cases the guidelines were not followed, we set up an inquiry among the 20 doctors involved in the follow-up in August 1998. RESULTS: The mean frequency of outpatient visits and physical examinations was 3-4 times per year, practically consistent with the guideline. Routine blood testing was performed in 17 patients (25%) and diagnostic imaging (X-ray or CT scan of the chest, ultrasonography of the liver) in 51 patients (76%) in deviation from the guideline. Non-compliance with the guideline could not be explained by unfamiliarity with the consensus, disagreement with the contents or existence of local protocols. Extra diagnostics were mostly meant to reassure patients. No metastases or recurrences were encountered during routine follow-up examinations, but some were found (in 8 patients) at interim visits to the outpatient clinic. CONCLUSION: The national guidelines regarding diagnostic tests in the follow-up of melanoma patients are insufficiently followed. Because redundant routine diagnostics probably have more disadvantages than benefits, a more active implementation of (future) guidelines appears necessary. PMID- 10368722 TI - [Intra-osseus gas formation in osteomyelitis of vertebrae and pelvis by Klebsiella pneumoniae]. AB - A 59-year-old woman had persistent sepsis after abdominal operations because of a volvulus and subsequently a retroperitoneal abscess, in spite of antibiotic treatment against Klebsiella pneumoniae, which grew in blood cultures. During abscess drainage, a haemorrhage from the infrarenal part of the aorta had occurred; in view of a presumed aortitis this part had been replaced by a bifemoral bypass. Computer tomography revealed intraosseous formation of gas in vertebrae and pelvis. At operation, abscesses were drained and necrotomy and sequestrotomy of the bone were performed. Cultures of the pus from the iliac crests showed K. pneumoniae. The antibiotic management was changed; the wounds were flushed regularly. After exposure of the wounds still draining after 6 months and vascular surgery because of occlusion of the bypass after 7 months, the patient recovered well. She died 2 years later from a cerebral haemorrhage. PMID- 10368723 TI - [Physical examination--rebound tenderness]. PMID- 10368724 TI - [Opportunistic screening for genital infections with Chlamydia trachomatis in sexually active population of Amsterdam. II. Cost-effectiveness analysis of screening women]. PMID- 10368726 TI - [Discordant fetal growth in multiple pregnancy: intervention should be based on chorionicity]. AB - In three women, aged 28, 35, and 38 years, with multiple pregnancies and discordant foetal growth, the question arose what to do in case of (threatening) intrauterine death of one twin. In one monochorionic pregnancy with single foetal death the survivor suffered irreversible neurological damage and died at the age of five months, in one monochorionic pregnancy the survivor was born healthy and in one dichorionic pregnancy both twins were born healthy although one twin showed severe intrauterine growth retardation. The problem concerning single foetal death in a monochorionic pregnancy is whether to terminate the pregnancy and accept the risk of premature birth to the surviving twin, or to continue the pregnancy and accept the risk of damage to the survivor. In a dichorionic pregnancy foetal death of one twin does not entail any great risk of damage to the survivor; in such a pregnancy single foetal death in a premature phase may be accepted and the pregnancy may be continued. Sonographic determination of the chorionicity in multiple pregnancy at an early stage is essential because it also determines the policy if foetal problems occur. PMID- 10368728 TI - [Immunology in medical practice. XXI. Laboratory tests for immunologic diseases]. AB - Clinical immunology has gained its firm place in health care. There are now established laboratory tests giving insight into the functioning of the immune system in the normal and diseased individual. The laboratory diagnostic tests are related to immune deficiencies, infectious diseases, allergic diseases, autoimmune diseases (both generalized and organ-specific ones), the HLA system and malignancies of immune cells. In this review the application of a number of laboratory tests in the diagnosis of immunological diseases is discussed. In the Netherlands a laboratory specialist, the 'medical immunologist' deals with the development, implementation and performance of the immunological diagnostic tests in health care. PMID- 10368727 TI - [Third circulation: twin transfusion syndrome]. AB - The estimated incidence in the Netherlands of serious previable twin-to-twin transfusion syndrome is 50-100 cases per year. The polyhydramnion-oligohydramnion sequence is the most prominent feature. Prognosis without treatment is dismal: previable preterm birth. Risk of serious morbidity in the surviving twin in case of foetal demise of the other is impressive. Once the diagnosis of monochorionicity has been made in the first trimester, detailed ultrasound examination is mandatory for early recognition of the development of the syndrome, as therapeutic options exist. Therapeutic options include piercing of the intertwin membrane, repeated amniocenteses or laser occlusion of the chorionic vasculature. In seven Dutch centres over three years time 61 cases were identified: 36 needed intervention; 63 (of the 122) children survived, 10 with longterm morbidity. PMID- 10368730 TI - [Roaming through methodology. XIII. Matching as a rule is not useful]. AB - In applied medical research matching is a technique used to control confounding in the design stage of a study. With matching, particular subjects are selected in such a way that the potential prognostic confounders are distributed equally among the comparison groups. A major disadvantage of matching is that it can be inefficient. With the so-called stratified data analysis control of confounding is achieved equally well. Therefore, from the validity and efficiency points of view matching is no longer desirable. PMID- 10368729 TI - [Informed well-considered consent ('informed consent'): an end or a means?]. AB - Since World War II, all sorts of protective measures have been taken for people who receive medical care or who are involved in scientific research. More and more, informed consent has become the standard. In the field of experimental therapeutical research, informed consent is still a controversial subject; the individual person's interests versus general interests. But informed consent appears to have become such an absolute prerequisite for all types of observational research that the future of epidemiological research based on existing data is threatened. According to both the Declaration of Helsinki and the Dutch Law for Protection of Persons, it is possible to omit asking individual informed consent under certain circumstances. Permission for research could then be given by a supervisory board or a medical-ethical commission. After all, informed consent was never meant to be a goal of its own, but a means for self protection and securing the right to autonomy. In situations where this right is curbed for other reasons, it appears that insisting on informed consent misses its target completely. PMID- 10368731 TI - [Cold tap water as a source of fatal nosocomial pneumonia due to Legionella pneumophila in a rehabilitation center]. AB - OBJECTIVE: Report of the technical, microbiological and epidemiological investigation following 2 cases of fatal Legionella pneumonia. DESIGN: Descriptive. METHOD: Faced with 2 nosocomial cases in a rehabilitation centre in the South of Limburg, the Netherlands, the water supply was investigated. Water temperatures from different taps were measured. Legionella cultures were made from respiratory patients' specimens, water samples and smears from all mixing taps (used in showers), samples from hot and cold water taps from the infected ward and from the five other wards. The strains were typed by serotyping and polymerase chain reaction. RESULTS: The circulating cold water sometimes warmed up to 40 degrees C (within the Legionella growth range). From the sputum of the 2 male patients with rheumatoid arthritis who died of Legionella pneumonia the same Legionella pneumophila (serotype I) was cultured as from the water supply. Of the showers on the contaminated ward 19% (12/63) were positive for Legionella as were 59% (35/59) of the cold water taps. Cultures from the hot water supply were negative just like control cultures from five other wards and swabs from showerheads and hoses. The cold water tubes ran next to the hot water tubes and the central heating system in the same shaft. On the infected ward patients were absent during the weekends. As one of the subsequent measures, the cold water pipes were relocated to another shaft. CONCLUSION: The combination of an elevated cold water temperature caused by heating along a distance by nearby hot water and heating piping and the regular stasis of water during the weekends when the ward was closed, most probably stimulated the multiplication of Legionella in the water supply. In order to minimize contamination of cold water its temperature must be kept below 20 degrees C. Surveillance of intramural water systems is necessary to prevent nosocomial infections. PMID- 10368733 TI - [Communicable diseases law: new legislation for infectious disease control]. AB - On April 1st, 1999 the new law on control of infectious diseases (the Infectious Diseases Act) came into effect. This law replaces former legislation, largely dating from 1928. The Infectious Diseases Act legally enables the government to take action against the spread of infectious diseases. This legal base is necessary because constitutional rights of individuals might be affected. The Infectious Diseases Act only applies to diseases specifically mentioned in the law. The Act regulates the statutory notification and the enforceable actions the authorities can take to protect the community. The mayors of municipalities decide on the appropriate measures, acting on advice of the municipal or regional public health agencies. The Infectious Diseases Act does not provide for detailed regulations regarding the control of infectious diseases but contains obligations that are the end piece of a policy continuum. PMID- 10368734 TI - [Roaming through methodology. XII. Pragmatic and pathophysiologic trials: a question of goal formulation]. PMID- 10368732 TI - [An epidemic of Salmonella typhimurium associated with traditional salted, smoked, and dried ham]. AB - OBJECTIVE: To discover the cause of an outbreak of gastroenteritis after a family party. DESIGN: Retrospective cohort study. METHODS: All 109 party-goers were asked to complete a written questionnaire about consumed food products and demographic and clinical variables and to hand in a faeces sample. The data were collected at the Public Health Institute Midden-Limburg, the Netherlands. Faeces and the remaining food products were examined microbiologically. The attack rates and the incidence rates of positive faeces culture among consumers and non consumers of specific food products were calculated as well as the corresponding relative risks (RR) with 95% confidence intervals (95% CI). RESULTS: The overall attack rate was 35%. Salmonella typhimurium phage type 20 was found in 'Coburger ham' and statistically significantly more frequently in faeces of ill compared with non-ill party-goers (RR: 6.4; 95% CI: 2.5-16.1). Twenty-eight different food products were served. Consumption of 'Coburger ham' only, was statistically significantly related to a positive faeces culture (RR: 4.1; 95% CI: 2.0-8.5). Only consumption of 'Coburger ham' and of 'bone ham' was statistically significantly related to being ill (RR: 2.4; 95% CI: 1.5-4.0 and RR: 1.4; 95% CI: 1.1-1.9, respectively). 'Coburger ham' and 'bone ham' originated from the same batch of raw meat and were prepared in the same manner in the same salt bath. The shorter duration of salting and drying of 'Coburger ham' compared with 'bone ham' corresponded with a higher relative risk of becoming ill. CONCLUSION: Consumption of 'bone ham' and 'Coburger ham' infected with S. typhimurium phage type 20 caused the outbreak. Traditional salting, drying and smoking of raw pork meat was not antimicrobiologically effective against S. typhimurium. Investigation of the antimicrobiological effect of the traditional preparation of meat and the importance of Good Manufacturing Practices and quality control in all stages of production of pork meat, according to the principles of Hazard Analysis and Critical Control Points, is advised. PMID- 10368735 TI - [A young child with fever of unknown origin; diagnosis and management]. PMID- 10368736 TI - [The young child with fever of unknown origin: diagnosis and management]. PMID- 10368737 TI - [The young child with fever of unknown origin: diagnosis and management]. PMID- 10368738 TI - [Vascular brain damage: a significant and identifiable cause of cognitive disorders and behavior disorders]. PMID- 10368739 TI - [Whipple's disease]. PMID- 10368740 TI - [Multiple amputations due to sepsis: however, functional rehabilitation is possible]. AB - Three patients had several major amputations because of disseminated intravascular coagulation accompanying purpura fulminans. A 31-year-old woman underwent a transfemoral amputation after a septic shock caused by haemolytic streptococcus A, which led to gangrene. A 57-year-old woman had a bilateral transtibial amputation after pneumococcaemia, and the third patient, a 37-year old woman, underwent a quadruple amputation following a meningococcal septic shock. The amputations were accompanied by contractures and skin damage due to ischaemic tissue changes. Additionally, cerebral and peripheral nerve dysfunction occurred. An intensive rehabilitation programme led to completely independent functioning with the use of orthotics and prosthetics. By starting a multidisciplinary approach as early as possible impairments can be treated properly and future disabilities minimized. PMID- 10368741 TI - [Sedation by non-anesthesiologists should be centralized for considerations of safety]. AB - Sedation and analgesia for diagnostic or therapeutic procedures outside the operating room by non-anaesthesiologist physicians is becoming more frequent. In reaction to sedation casualties a multidisciplinary committee organized by the National Organization for Quality Assurance in Hospitals (CBO) has developed guidelines for sedation and analgesia by non-anaesthesiologists for psychologically or physically distressing diagnostic and therapeutic procedures. In these guidelines the conditions for a qualitatively safe way of sedation or analgesia are being emphasized. It may be expected that with these guidelines sedation and analgesia by non-anaesthesiologists will increase further. With a view to the safety of the patient diagnostic and therapeutic procedures which require sedation for psychological or physical reasons should be concentrated in a diagnostic/therapeutic complex, connected to the operating complex, where it is possible to consult the expertise of an anaesthesiologist. PMID- 10368742 TI - [Epidemic of cardiovascular diseases in patients undergoing chronic hemodialysis; closer attention needs to be paid to volume expansion]. AB - The very high mortality of patients on chronic haemodialysis treatment is mainly due to cardiovascular complications. Despite the fact that some dialysis centres report much better survival by adhering to the long-known principle of volume control, investigators and advisory committees all over the world define 'adequate dialysis' exclusively in terms of removal of 'toxic factors'. Volume expansion leads to hypertension, cardiac dilatation and cardiac failure through a clear pathophysiological sequence. By giving more responsibility to dialysis nurses patients may be helped to regain their 'dry weight'. PMID- 10368743 TI - [Cardiovascular diseases in dialysis patients]. AB - Cardiovascular morbidity and mortality are higher among dialysis patients than among the general population. The cardiovascular problems often exist before the start of dialysis. Their pathogenesis is multifactorial in dialysis patients also. The prevalence of left ventricular hypertrophy is strongly increased. Adequate therapy may lead to partial remission. Cardiac ischaemia is frequent among dialysis patients and may occur without severe coronary artery disease. The prognosis of myocardial infarction in dialysis patients is poorer than in the general population. There is no proven difference between the various dialysis techniques regarding cardiovascular morbidity and mortality, while kidney transplantation may have a beneficial effect. Early diagnosis and treatment aimed at risk factors for cardiovascular disease are indicated. PMID- 10368744 TI - [Contrast-enhanced magnetic resonance angiography]. AB - Contrast-enhanced magnetic resonance angiography (MRA) involves intravenous injection of a contrast medium that increases the signal intensity of blood by shortening its T1 value. With contrast-enhanced MRA the acquisition time is short (less than 40 s for the abdominal aorta and the iliac vessels) and the images obtained can be interpreted accurately. The contrast medium currently in use virtually never causes adverse effects and is not nephrotoxic. After obtaining a three-dimensional dataset projections can be made at will. In addition, the individual partitions should be evaluated. The postprocessing time is about 15 min per examination. Current clinical applications are diagnostic examination of (stenoses of) the aortic arch and its branches, the thoracic and abdominal aorta, the visceral vessels, the renal arteries and the peripheral arteries. The sensitivity and specificity of contrast-enhanced MRA in most studies amount to over 90%. PMID- 10368745 TI - [Arterial perfusion disorders of the hand in 9 patients with arteriovenous fistula for hemodialysis]. AB - OBJECTIVE: To describe the treatment of hand ischaemia as a complication of arteriovenous fistulas, which have been used for haemodialysis vascular access. DESIGN: Retrospective. METHODS: In 1990-1998 there were in the University Hospital Maastricht, (AZM), the Netherlands, 9 patients with hand ischaemia as a complication of arteriovenous fistulas for haemodialysis. Five of these originated from the AZM, where in the same period 341 fistulas had been created. Data were collected from archives about the treatment in the 9 patients and its results. RESULTS: The patients were 7 women and 2 men, with a mean age of 61 years (range: 35-73). Four patients had diabetes mellitus. All patients had a high-flow AV fistula (mean flow: 1556 ml/min) at the level of the elbow. Surgical revision to diminish access flow volume was carried out in 2 patients, while fistula closure with creation of a new AV fistula was performed in 7 patients. Because of persistent ischaemia in 2 patients the access site was closed. Finger amputation was necessary in 3 patients. CONCLUSION: Of all patients with new vascular accesses 1.5% developed symptomatic ischaemic complications. Two out of 9 AV fistulas could be preserved after surgical correction and in 3 patients finger amputations were carried out because of irreversible necrosis. PMID- 10368747 TI - [Malignant hypertension in a young man with renal artery occlusion diagnosed with magnetic resonance angiography]. AB - A male aged 22 years developed a hypertensive crisis with encephalopathy after his antihypertensive medication had been discontinued with a view to extended diagnostics. Immediate intensive treatment led to rapid and complete recovery. By using gadopentetate acid enhanced magnetic resonance angiography it is possible to obtain a clear image of the morphology of the kidneys and the renal vasculature without the use of iodinated contrast media and arterial catheterisation. This technique revealed an occluded renal artery and a recent infarction that possibly had led to the serious and threatening events. PMID- 10368746 TI - [Guideline for administration of sedatives and analgesics by physicians who are not anesthesiologists. National Organization for Quality Assurance in Hospitals]. AB - A consensus text for sedation or analgesia in diagnostic or therapeutic procedures has been developed for application by non-anaesthetist physicians. The final consensus text has the support of 17 scientific societies in the Netherlands. There is not enough medical manpower for direct, personal, specialist-based supervision of level 3 sedation procedures (the patient is relaxed, with eyes closed, but promptly reacts to verbal commands) for significant number of patients in the Netherlands. Sedation and analgesia may be administered by other health care personnel than anaesthesiologists when a number of conditions are met. These concern risk management, information to the patient, patient's consent, requirements for medical and supporting personnel, equipment, sedation procedures, monitoring, data management, recovery and aftercare. The consensus party favours titrated administration of small doses of short acting sedative or analgetic drugs. Combining sedative and analgesic drugs increases risk. Sedation and analgesia in children and patients with mental handicaps is acceptable in terms of quality, but requires special expertise, because of the greater psychic and physical vulnerability of these categories of patients. PMID- 10368748 TI - [Risk factors for dying among ibopamine users]. PMID- 10368749 TI - [Informed well-considered consent ('informed consent'): an end or a means?]. PMID- 10368750 TI - [Intracranial arteriovenous malformations in pregnant women]. PMID- 10368751 TI - [Analysis of chromatin conformational flexibility in drosophila embryo cell free system]. PMID- 10368752 TI - [The difference in the mechanism of irreversible inhibition of cholinesterases from different species]. PMID- 10368753 TI - [The export and absorption of the prosome-like RNP (alphaRNP) displaying the activity of immunomodulation in cell cultures]. PMID- 10368754 TI - [Specific proteins of the lens epithelium of amphibian eyes]. PMID- 10368755 TI - [Age-dependent changes in brain monoamine oxidase in Brattleboro rats with genetically determined diabetes insipidus]. PMID- 10368756 TI - [The reactions of chipmunk binocular striatal cortex on performed stimuli]. PMID- 10368757 TI - [Ca-ATPase of plasma membrane--molecular target for metastazin]. PMID- 10368758 TI - [Combined inhibition of monoamine oxidase]. PMID- 10368759 TI - [The effect of chromatin nonhistone protein from rat brain on early embryogenesis of mice]. PMID- 10368760 TI - [Abnormal structure of blood cells and related hemostasis reactions during persistence of mycoplasma infection in humans]. PMID- 10368761 TI - [Comparative analysis of development of the auditory and vestibular structures in a representative of true seals, Pusa hispida (Pinnipedia:Phocidae-Pusa hispida)]. PMID- 10368762 TI - [Analysis of chromatin structural changes in Drosophila embryo cell free system]. PMID- 10368763 TI - [Self-assembly of soluble X-oligomers]. PMID- 10368764 TI - [Tissue- and species-specific markers of hematopoietic cells from rodent bone marrow]. PMID- 10368765 TI - [Activating effect of 4-aminopyridine on inward rectifier potassium current in embryonic skeletal frog myocytes]. PMID- 10368766 TI - [The role of the quantitative factor in alteration of population functional status during normal and neoplastic growth]. PMID- 10368767 TI - [Specificity of biochemical reactions of Xenopus laevis to oxidative stress after preceding action by hyperbaric oxygenation]. PMID- 10368768 TI - [Hearing sensitivity to contrast of sound spectrum patterns]. PMID- 10368769 TI - Neuroendocrine influences and repercussions of the menopause. AB - In summary, the evidence that both the ovary and the brain are key pacemakers in the menopause is compelling. Our appreciation that estrogens are important neurotrophic and neuroprotective factors has grown rapidly. Future studies will allow us to better understand the ensemble of factors that interact to maintain regular reproductive cyclicity and how this precise dynamic balance changes with age. Furthermore, understanding how estrogen exerts trophic and protective actions should lead to its use as an important therapeutic agent in the maintenance of normal neural function during aging and after injury. PMID- 10368770 TI - Assisted reproduction-in vitro fertilization success is improved by ovarian stimulation with exogenous gonadotropins and pituitary suppression with gonadotropin-releasing hormone analogues. PMID- 10368771 TI - Tamoxifen, raloxifene, and the prevention of breast cancer. PMID- 10368772 TI - Estrogen actions in the central nervous system. PMID- 10368773 TI - Alternatives to the use of estrogen in postmenopausal women. PMID- 10368774 TI - Nuclear receptor coregulators: cellular and molecular biology. AB - Nuclear receptor coregulators are coactivators or corepressors that are required by nuclear receptors for efficient transcripitonal regulation. In this context, we define coactivators, broadly, as molecules that interact with nuclear receptors and enhance their transactivation. Analogously, we refer to nuclear receptor corepressors as factors that interact with nuclear receptors and lower the transcription rate at their target genes. Most coregulators are, by definition, rate limiting for nuclear receptor activation and repression, but do not significantly alter basal transcription. Recent data have indicated multiple modes of action of coregulators, including direct interactions with basal transcription factors and covalent modification of histones and other proteins. Reflecting this functional diversity, many coregulators exist in distinct steady state precomplexes, which are thought to associate in promoter-specific configurations. In addition, these factors may function as molecular gates to enable integration of diverse signal transduction pathways at nuclear receptor regulated promoters. This review will summarize selected aspects of our current knowledge of the cellular and molecular biology of nuclear receptor coregulators. PMID- 10368775 TI - Modulation of osteoclast differentiation and function by the new members of the tumor necrosis factor receptor and ligand families. AB - Osteoblasts/stromal cells are essentially involved in osteoclast differentiation and function through cell-to-cell contact (Fig. 8). Although many attempts have been made to elucidate the mechanism of the so-called "microenvironment provided by osteoblasts/stromal cells," (5-8) it has remained an open question until OPG and its binding molecule were cloned. The serial discovery of the new members of the TNF receptor-ligand family members has confirmed the idea that osteoclast differentiation and function are regulated by osteoblasts/stromal cells. RANKL, which has also been called ODF, TRANCE, or OPGL, is a member of the TNF ligand family. Expression of RANKL mRNA in osteoblasts/stromal cells is up-regulated by osteotropic factors such as 1 alpha, 25(OH)2D3, PTH, and IL-11. Osteoclast precursors express RANK, a TNF receptor family member, recognize RANKL through cell-to-cell interaction with osteoblasts/stromal cells, and differentiate into pOCs in the presence of M-CSF. RANKL is also involved in the survival and fusion of pOCs and activation of mature osteoclasts. OPG, which has also been called OCIF or TR1, is a soluble receptor for RANKL and acts as a decoy receptor in the RANK-RANKL signaling system (Fig. 8). In conclusion, osteoblasts/stromal cells are involved in all of the processes of osteoclast development, such as differentiation, survival, fusion, and activation of osteoclasts (Fig. 8). Osteoblasts/stromal cells can now be replaced with RANKL and M-CSF in dealing with the whole life of osteoclasts. RANKL, RANK, and OPG are three key molecules that regulate osteoclast recruitment and function. Further studies on these key molecules will elucidate the molecular mechanism of the regulation of osteoclastic bone resorption. This line of studies will establish new ways to treat several metabolic bone diseases caused by abnormal osteoclast recruitment and functions such as osteopetrosis, osteoporosis, metastatic bone disease, Paget's disease, rheumatoid arthritis, and periodontal bone disease. PMID- 10368776 TI - Estrogen receptor null mice: what have we learned and where will they lead us? AB - All scientific investigations begin with distinct objectives: first is the hypothesis upon which studies are undertaken to disprove, and second is the overall aim of obtaining further information, from which future and more precise hypotheses may be drawn. Studies focusing on the generation and use of gene targeted animal models also apply these goals and may be loosely categorized into sequential phases that become apparent as the use of the model progresses. Initial studies of knockout models often focus on the plausibility of the model based on prior knowledge and whether the generation of an animal lacking the particular gene will prove lethal or not. Upon the successful generation of a knockout, confirmatory studies are undertaken to corroborate previously established hypotheses of the function of the disrupted gene product. As these studies continue, observations of unpredicted phenotypes or, more likely, the lack of a phenotype that was expected based on models put forth from past investigations are noted. Often the surprising phenotype is due to the loss of a gene product that is downstream from the functions of the disrupted gene, whereas the lack of an expected phenotype may be due to compensatory roles filled by alternate mechanisms. As the descriptive studies of the knockout continue, use of the model is often shifted to the role as a unique research reagent, to be used in studies that 1) were not previously possible in a wild-type model; 2) aimed at finding related proteins or pathways whose existence or functions were previously masked; or 3) the subsequent effects of the gene disruption on related physiological and biochemical systems. The alpha ERKO mice continue to satisfy the confirmatory role of a knockout quite well. As summarized in Table 4, the phenotypes observed in the alpha ERKO due to estrogen insensitivity have definitively illustrated several roles that were previously believed to be dependent on functional ER alpha, including 1) the proliferative and differentiative actions critical to the function of the adult female reproductive tract and mammary gland; 2) as an obligatory component in growth factor signaling in the uterus and mammary gland; 3) as the principal steroid involved in negative regulation of gonadotropin gene transcription and LH levels in the hypothalamic pituitary axis; 4) as a positive regulator of PR expression in several tissues; 5) in the positive regulation of PRL synthesis and secretion from the pituitary; 6) as a promotional factor in oncogene-induced mammary neoplasia; and 7) as a crucial component in the differentiation and activation of several behaviors in both the female and male. The list of unpredictable phenotypes in the alpha ERKO must begin with the observation that generation of an animal lacking a functional ER alpha gene was successful and produced animals of both sexes that exhibit a life span comparable to wild-type. The successful generation of beta ERKO mice suggests that this receptor is also not essential to survival and was most likely not a compensatory factor in the survival of the alpha ERKO. In support of this is our recent successful generation of double knockout, or alpha beta ERKO mice of both sexes. The precise defects in certain components of male reproduction, including the production of abnormal sperm and the loss of intromission and ejaculatory responses that were observed in the alpha ERKO, were quite surprising. In turn, certain estrogen pathways in the alpha ERKO female appear intact or unaffected, such as the ability of the uterus to successfully exhibit a progesterone-induced decidualization response, and the possible maintenance of an LH surge system in the hypothalamus. [ABSTRACT TRUNCATED] PMID- 10368777 TI - Selective estrogen receptor modulators: clinical spectrum. PMID- 10368778 TI - [Current data on bipolar disorders, epilepsy and spasticity. 4. Neuro-Forum, Dresden]. PMID- 10368780 TI - [Evolution of differential chromosome banding]. AB - Specific chromosome banding patterns in different eukaryotic taxons are reviewed. In all eukaryotes, chromosomes are composed of alternating bands, each differing from the adjacent material by the molecular composition and structural characteristics. In minute chromosomes of fungi and Protozoa, these bands are represented by kinetochores (Kt- (Cd-)bands), nucleolus organizers (N-bands), and telomeres as well as the euchromatin. In genomes of most fungi and protists, long clusters of tandem repeats and, consequently, C-bands were not revealed but they are likely to be found out in species with chromosomes visible under a light microscope, which are several tens of million bp in size. Chromosomes of Metazoa are usually larger. Even in Cnidaria, they contain C-bands, which are replicated late in the S phase. In Deuterostomia, chromosome euchromatin regions differ by replication time: bands replicating at the first half of the S phase alternate with bands replicating at the second half of the S phase. Longitudinal differentiation in the replication pattern of euchromatic regions is observed in all classes of Vertebrata beginning with the bony fish although the time when it developed in Deuterostomia is unknown. Apparently, the evolution of early and late replicating subdomains in Vertebrata euchromatin promoted fast accumulation of differences in the molecular composition of nucleoproteid complexes characteristic of early and late replicating bands. As a result, the more contrasting G/R and Q-banding patterns of chromosomes developed especially in Eutheria. The evolution of Protostomia and Plantae followed another path. An increase in chromosome size was not accompanied by the appearance of wide RBE and RBL euchromatin bands. The G/R-like banding within the interstitial chromosome regions observed in some representatives of Invertebrates and higher plants arose independently in different phylogenetic lineages. This banding pattern seems to be closer to that of C-banding than to the typical G/R-banding of the mammalian chromosomes. PMID- 10368779 TI - [Various forms of risk factors of antidepressive agents for influencing sexual functions. Moclobemid is first choice for the therapy of sexually active patients with depression]. PMID- 10368781 TI - [x811--mutation of the Pseudomonas aeruginosa transposable phage D3112 with a pleiotropic effect]. AB - Characteristics of Pseudomonas aeruginosa Transposable Phage D3112 carrying mutation x811 are described. x811 is a recessive mutation with pleiotropic effect. It determines a deteriorated lysis of infected or induced bacteria, a delayed replication, and a considerably decreased replication rate. In addition, the x811 mutation is expressed as the Kil phenotype, since high-temperature induction of prophage D3112 cts15 x811 does not cause an immediate decrease in the ability of bacteria to form colonies at 42 degrees C. Restriction analysis of DNA of D3112 cts15 x811 and its segregants has not revealed extended insertions or deletions. The characteristics of segregants of the D3112 cts15 x811 phage agree with the suggestion that the x811 mutation has emerged in a regulatory element (a gene or a site) that controls both expression of the entire early operon, including the "pre-early" function Kil, and the regulation of the repressor synthesis. PMID- 10368782 TI - [Natural plasmid pSD89 (Cmr) from Salmonella derby K89, which increases the radiation tolerance of Escherichia coli strain K-12]. AB - Several plasmids with molecular mass of 1.3-9 MDa were found in a clinical isolate of Salmonella derby K89 by electrophoresis in agarose gel. One of these plasmids, designated pSD89 (Cmr), was derived from the K89 strain via transformation of the plasmidless recipient S. derby K82 to chloramphenicol resistance. The plasmid-carrying strain K89 and the K82 strain completely cured of plasmids were equally sensitive to the lethal action of UV light, whereas the plasmid-carrying strain was even more sensitive to ionizing radiation than the plasmidless variant. Nevertheless, transformants carrying only plasmid pSD89 (Cmr) were found to be more resistant to gamma-rays and UV light than the recipient. By using an intermediate host Escherichia coli Z80 (r-m+), plasmid pSD89 (Cmr) was introduced into different E. coli K-12 strains: polA-, recA-, uvrA-, umuC-, and the wild-type strain. A slight increase in radioresistance of E. coli wild-type cells and a significant complementation of a repair defect in recA and polA mutants, but not in uvrA and umuC, were observed. PMID- 10368783 TI - [A mutant allele gam18, participating in the RecF repair path in Escherichia coli K-12]. AB - Plasmid pGam18 carrying one of the cloned mutant loci, responsible for enhanced radiation resistance in the strain Escherichia coli Gamr444, was shown to increase resistance to the lethal effect of gamma-rays with a dose modification factor DMF = 2. Enhanced resistance was observed in wild-type cells and in the mutant recBC sbcB, but not recFBC sbcA. This indicates the involvement of a product of the gam18 locus in the RecF pathway of recombinational repair. The protective effect of plasmid pGam18 against radiation was completely abolished by mutations in the most RecF pathway genes (recF, recJ, recR, recO, recQ, recN, and ruvB). However, three mutations in the uvrD gene, which encodes DNA helicase II and belongs to the RecF pathway, can be partially complemented by plasmid pGam18. These data suggest that the mutant allele gam18 affects the DNA helicase II activity at the presynaptic stage of the RecF pathway-mediated repair of DNA double-stranded breaks induced by gamma-irradiation. PMID- 10368784 TI - [Cloning and expression of the gene for the mycoplasma key division protein FtsZ in Escherichia coli]. AB - Gene ftsZ responsible for division of bacterial cells was revealed in most prokaryote groups. A 520-bp fragment of the ftsZ gene was amplified on the template of A. laidlawii DNA using degenerate primers. This fragment was sequenced and served as a hybridization probe for cloning of the full-sized copy of the A. laidlawii ftsZ gene. The amplified fragment was cloned in a pGEX3X vector and expressed in E. coli cells. Polyclonal antibodies derived from the chimeric polypeptide containing a fragment of A. laidlawii FtsZ protein interacted only with the A. laidlawii protein with molecular mass of 40 kDa. Comparison of nucleotide sequences of the ftsZ-gene region of A. laidlawii and other bacterial species showed that they were highly homologous in A. laidlawii, E. coli, and Bac. subtilis, while low homology was revealed between the A. laidlawii sequence and those of the members of the genus Mycoplasma. Analysis of the ftsZ-gene nucleotide sequences is suggested as a means to study the evolutionary relatedness of prokaryotes. PMID- 10368785 TI - [Genetic and biological characteristics of noninbred mice from the ICR colony]. AB - Noninbred mice of the ICR colony were studied by a set of characters making it possible to estimate the range of genetic polymorphism in the given population. Genotypes of mice for loci A-, B-, D-, S-, PP, Se Se, and cc were determined. Noninbred mice were polymorphic for loci A, B, D, and S. Frequencies of recessive alleles of loci A, B, and D was calculated. Noninbred mice have a high fecundity and a low level of embryonic mortality, which indicate population heterogeneity. The age of the mother was shown to have no effect on the ovulation norm. The population standard of the colony for age-dependent change in the body weight and size was established. Age-dependent survival in females and males was shown to be associated. PMID- 10368786 TI - [Linkage of a porcine esterase D locus with a lethal factor]. AB - This work is devoted to a linkage study of the gene controlling esterase D and a lethal factor in domestic pig. Segregation for two alleles EsDB, which originated from populations of miniature swine and pigs of a large white breed, and the effect of these variants on the number of offspring in a litter were compared. The results showed a linkage of locus EsD with a lethal factor. This lethal factor causes the mortality of homozygotes and half of the heterozygotes in a litter, which depends on the litter size. The lethal factor was found to be absent in miniature pigs, and in the large white breed it is predominantly linked with allele EsDB. The effect of the lethal factor on population structure and fecundity is discussed. PMID- 10368787 TI - [Allozyme differentiation of various chromosomal forms of the spotted suslik (Spermophilus suslicus Guld, 1770, Rodentia)]. AB - Eleven enzyme and four nonenzyme protein systems controlled by 25 loci were electrophoretically analyzed in the allopatric karyotypic forms (2n = 34 and 2n = 36) of the spotted souslik Spermophilus suslicus. Genetic variability and differentiation for the forms with different chromosome sets were estimated. Two discriminative loci (Alb and Tf) for the studied chromosome forms were found. The UPGMA dendrogram was constructed, which summarizes genetic (allozyme) relationships found for the forms of the spotted souslik with different chromosome sets. Subdividing the species into two karyotypic forms was shown to be followed by differentiating these forms at the allozyme level. PMID- 10368788 TI - [Burden of hereditary diseases in residents of the Mari El Republic]. AB - A summary of the medical genetic studies of the Marii El population is presented. A total of 276,900 people, 110,894 and 166,006 urban and rural inhabitants, respectively, were examined. Regarding the ethnic composition, the studied population was mostly Mari (61.96%) and Russian (32.04%). Medical genetic examination revealed 480 subjects from 260 families with autosomal dominant (AD) diseases, 234 subjects from 184 families with autosomal recessive (AR) diseases, and 49 subjects from 41 families with x-linked diseases. Segregation analysis revealed a good agreement between the expected and observed segregation frequencies for families with AR and AD diseases and allowed the frequency of hereditary diseases in the urban and rural, as well as the Russian and Mari, populations, to be estimated. The total frequency of AD diseases in Maris was approximately twice as high as in Russians (1.99 and 0.97%, respectively); substantial differences between district populations were found. The total frequency of AR diseases was also two times higher in Maris than in Russians (1.00 and 0.54%, respectively). The frequencies of AR and AD diseases in different districts were correlated with the levels of random and local inbreeding, population size, and the index of maximum selection. PMID- 10368789 TI - [Statistical methods of radiation hybrid mapping: analysis of a small number of genes]. AB - A statistical method for radiation hybrid (RH) mapping of a small number of genes detected in clones by means of isozyme analysis is suggested. This method allows analytical solution of the problem of gene ordering and qualitative comparison of relative distances between the genes that are being mapped. The possibilities offered by the suggested method are illustrated using earlier data on the distribution of four isozyme markers in human and vole (Microtus rossiaemeridionalis) clones. These markers are hypoxanthine phosphoribosyl transferase (HPRT), PGK, G6PD, and GALA. PMID- 10368790 TI - [Analysis of an operator-like structure, regulating the activity of the ribC gene in Bacillus subtilis]. AB - A point mutation (C-->A substitution) in the -35 region of a putative promoter operator site TTGCCG-17n-TACATT results in a more than 25-fold increase in the activity of ribC gene encoding the bifunctional enzyme--flavokinase/FAD-synthase- in Bacillus subtilis. PMID- 10368791 TI - [Detection of a new tissue-specific enhancer in the ct6 region of the regulatory region of the drosophila cut locus]. AB - The cut locus of Drosophila is an interesting example of a complex eukaryotic locus responsible for the development of many tissues and organs. Most of this locus is regulatory. The entire locus was cloned by Tchurikov et al. in 1986 and Blochlinger et al. in 1988. The wing ctn enhancer located 80 kb upstream of the promoter was earlier found in a 2.7 kb EcoRI-BamHI DNA fragment. The locus region 65-80 kb remote from the promoter was assumed to control the development of wings and vibrissae. We have found a new enhancer region in the ct6 region of the locus, which was in a 5 kb BamHI-EcoRI DNA fragment adjacent to the ctn enhancer. This region is responsible for the expression of the reporter lacZ gene in many tissues and organs at all stages of Drosophila development (at least in the intestine, Malpighian tubules, thoracic and abdominal sensory organs, thoracic ganglia and in ring glands). Thus, the region located 75 kb upstream of the promoter has some properties of the locus control region (LCR). PMID- 10368792 TI - Gender-specific medicine: the new profile of gynecology. AB - The science of gynecology is undergoing a change and is swiftly turning into a holistic discipline, i.e. gender-specific medicine. The rationale for this is that the hormones of the ovary not only are responsible for reproduction but also perform a number of extragenital functions that extend far into other disciplines, giving rise to a different frequency of diseases in women than in men. For example, females are five times more likely to be affected by rheumatoid arthritis than males, the same also holding true for autoaggressive conditions. This phenomenon may be accounted for by the fact that physiological auto aggression is involved in the reproductive process. Similarly, there is a difference between women and men with regard to the sicca phenomenon, or to such disorders as connective tissue weakness, cellulite, venous conditions or hypercholesterolemia. A cause-related treatment of such problems is now available through specific endocrine therapy. That is why gynecologists in future will increasingly have to adopt an interdisciplinary approach. PMID- 10368793 TI - Increase of axial and appendicular trabecular and cortical bone density in established osteoporosis with intermittent nasal salmon calcitonin therapy. AB - The aim of this study was to examine the effect of intranasal administration of salmon calcitonin to a group of 24 postmenopausal women with severe, established osteoporosis (t score < -2.5 SD) and more than one vertebral fracture. The patients were treated with 200 IU of nasal salmon calcitonin daily for 2 months with a subsequent pause of 2 months (3 cycles) and 500 mg calcium daily over a total of 12 months in an open randomized study. The patients were compared with an age matched control group of 18 women of a similar clinical status who were treated with calcium and vitamin D only. In the nasal calcitonin treatment group an increase in the trabecular axial bone density of 2.8% was achieved, as well as increase in trabecular appendicular (forearm) bone density of 1.6%, together with a cortical bone density increase of 1.8% axial and 1% appendicular. Initially, elevated values of urinary deoxypyridinoline were found in 12 women in the nasal calcitonin treatment group; these levels returned to normal under salmon calcitonin nasal therapy and documented the inhibition of increased osteoclastic activity. Cyclic intermittent calcitonin nasal therapy led to a general increase in trabecular and cortical axial and appendicular bone density, marked alleviation of the subjective sensation of pain, and a reduction in the daily dose of accompanying nonsteroidal anti-inflammatory drugs by 50%. PMID- 10368794 TI - Endocrine, neuroendocrine and behavioral effects of oral dehydroepiandrosterone sulfate supplementation in postmenopausal women. AB - Aging in women and men is characterized by a progressive decline of circulating dehydroepiandrosterone (DHEA) levels and its sulfate ester (DHEAS). The improvement of wellbeing described in postmenopausal women treated with DHEA suggests that this steroid may exert specific actions on the central nervous system (CNS). The postmenopausal period is associated with several neuroendocrine modifications. The decrease of circulating levels of beta-endorphin is considered a hormonal marker of those changes. The aim of the present study was to investigate neuroendocrine and behavioral effects of three months of DHEAS supplementation in postmenopausal women. Postmenopausal women (n = 22) were divided in three groups: the first group was treated with oral DHEAS (n = 8) (50 mg/day), the second treated with the same dose of oral DHEAS + transdermal estradiol (n = 8) (DHEAS) 50 mg/day, estradiol 50 micrograms/patch) and the third with transdermal estradiol alone (n = 6) (50 micrograms/day). Before and after 1, 2 and 3 months of therapy, the following circulating steroid and protein hormone levels were evaluated: DHEA, DHEAS, androstenedione, testosterone, estrone, estradiol, 17-hydroxyprogesterone, sex hormone-binding globulin (SHBG), follicle stimulating hormone (FSH), luteinizing hormone (LH), beta-endorphin, growth hormone (GH) and cortisol, and a Kupperman score was performed. Before and after treatments, plasma beta-endorphin levels were evaluated in response to three neuroendocrine tests: (a) clonidine, an alpha 2-presynaptic adrenergic agonist (1.25 mg i.v.) (b) naloxone, an opioid receptor antagonist (4 mg i.v.) and (c) fluoxetine, a serotonin selective reuptake inhibitor (30 mg p.o.). In both groups of women treated with DHEAS, mean basal serum DHEA, DHEAS, androstenedione, and testosterone levels significantly increased after treatment, while no changes were shown in the group receiving estradiol alone. Serum estradiol, estrone, GH and plasma beta-endorphin levels significantly increased progressively for the three months of treatment, with higher levels for estrone and estradiol in subjects receiving estradiol alone or plus DHEAS. Serum SHBG, cortisol, and 17 hydroxyprogesterone did not show significant variations under any treatment. Serum LH and FSH levels showed a significant decrease in groups treated with estradiol alone or plus DHEAS at the second and third months. The Kupperman score showed that all treatments were associated with similar and progressive improvement. Before therapy clonidine, naloxone and fluoxetine stimuli failed to modify circulating beta-endorphin levels. After each of the treatments, the beta endorphin response was completely restored and was similar, independent of the kind of therapy. Restoration of the beta-endorphin response to specific stimuli suggests that DHEAS and/or its active metabolites modulates the neuroendocrine control of pituitary beta-endorphin secretion, which may support the therapeutic efficacy of the DHEAS on behavioral symptoms. PMID- 10368795 TI - Circulating levels of immunoreactive beta-endorphin in polycystic ovary syndrome. AB - The plasma levels of beta-endorphin were studied in 64 women with polycystic ovarian disease (PCOD), from whom was selected a group of 23 women with normal weight and amenorrhea of < 36 days. On day 21, beta-endorphin levels were: mean 64.92 pg/ml; SD 37.32 pg/ml; 95% CI 48.38-81.47 pg/ml. It was also observed that their levels of opioid peptide were reduced, compared with women who had normal ovulatory cycles, both in the follicular phase (mean 70.93 pg/ml; SD 24.59 pg/ml; 95% CI 76.84-99.77 pg/ml) and luteal phase (mean 88.30 pg/ml; SD 31.80 pg/ml; 95% CI 76.84-99.77 pg/ml). The results were statistically significant (p < 0.05) for levels in PCOD patients compared with those of the luteal phase in women with normal ovulatory cycles. The decreased levels of beta-endorphin were negatively related to luteinizing hormone (LH) levels, which might explain the rise of LH levels in women with PCOD who control their weight and at the time of amenorrhea, although it is not clear if central opioid activity is reflected in the peripheral blood. PMID- 10368796 TI - Serum leptin concentrations in obese women with Down syndrome and Prader-Willi syndrome. AB - We have evaluated serum leptin concentrations in two forms of genetic obesity. The subjects examined were eight women with Down syndrome and eight women with Prader-Willi syndrome. All patients were in the reproductive age range and were obese (body mass index > or = 27 kg/m2). Plasma leptin values, analyzed as a function of body mass index showed a statistically significant correlation in both Prader-Willi (r = 0.985; p < 0.001) and Down syndrome patients (r = 0.943; p < 0.001). Obese Down syndrome women exhibited significantly lower leptin values (10.8 +/- 1.1) as compared to patients with Prader-Willi syndrome (31 +/- 2.6; p < 0.01). The linear correlation between leptin and insulin in the two groups of patients was not statistically significant. The data suggested that obesity in Prader-Willi subjects could be caused by failure of leptin to reach its target in the brain, as a consequence of defects in the receptor or in postreceptor processing, whereas data on obese patients with Down syndrome could be due to a different pathogenetic origin. PMID- 10368797 TI - Diagnosis of subtle ovulation disorders in subfertile women with regular menstrual cycles: cost-effective clinical practice? AB - Serial monitoring by plasma progesterone measurement is advised in the literature for fertility work-up, to detect ovulation disturbances in women presenting with regular menstrual cycles. Three strategies to diagnose such 'subtle ovulation disorders' (SOD, defined as anovulation, inadequately timed ovulation or ovulation of a follicle of reduced size in regularly cycling women) were evaluated, in order to investigate costs of such a diagnosis. On the basis of a 'maximal', an 'ultrasound-only', and a 'preselection' strategy, total medical costs and costs including non-medical costs were calculated for each SOD diagnosis. A 'maximal' diagnostic strategy resulted in a total medical cost of ECU 9057 per diagnosis (including non-medical costs ECU 12,787); an 'ultrasound only' strategy in ECU 4520 (ECU 6791) per diagnosis. By use of a 'preselection' strategy, 4.25% of the women were found to have an SOD, at a cost of ECU 3036 (ECU 6868) for each diagnosis. As the real significance of SOD diagnosis for the prognosis of the patient to become pregnant without treatment remains unclear, and as no randomized trials on treatment effectiveness have as yet been undertaken, it is questionable whether this approach is worthwhile. PMID- 10368798 TI - The treatment of severe premenstrual syndrome with goserelin with and without 'add-back' estrogen therapy: a placebo-controlled study. AB - The study aimed to determine if the addition of daily low-dose oral estrogen with a cyclical progestogen given to young women using a depot gonadotropin-releasing hormone (GnRH) analog implant for the treatment of their premenstrual syndrome (PMS) would affect the clinical outcome. In a double-blind placebo-controlled study in a specialist premenstrual syndrome clinic setting, 60 women aged between 20 and 45 years were randomized to one of three treatment groups: Group A (placebo implant four weekly + placebo tablets daily), Group B (goserelin 3.6 mg implant four weekly + estradiol valerate 2 mg daily with norethisterone 5 mg from days 21-28 of a 28-day cycle) or Group C (goserelin 3.6 mg implant four weekly + placebo tablets daily). Differences between PMS scores at 2, 4 and 6 months were compared with pretreatment values. There was a significant improvement in PMS scores in Group C (Zoladex + placebo) after 2, 4 and 6 months of treatment when compared to pretreatment values and Group A (placebo + placebo). The addition of a low-dose oral estrogen with a cyclical progestogen to GnRH analog treatment (Group B) resulted in a less dramatic response when compared to pretreatment values and no significant improvement when compared to Group A (placebo + placebo) at 2, 4 and 6 months of treatment. The addition of a low-dose oral estrogen with a cyclical progestogen to depot GnRH analog therapy in the treatment of PMS reduces the clinical response. PMID- 10368799 TI - Renin-angiotensin systems and reproduction. PMID- 10368800 TI - [The role of putrescine and the energy state of Escherichia coli in regulation of DNA topology during adaptation to oxidative stress]. AB - An exponential-phase culture of E. coli responded to the addition of H2O2 by a decrease in DNA supercoiling induced by the lowering of the energy status of cells, potassium leakage, and breaking of polynucleotide chains. Extending the time of exposure of E. coli cells to hydrogen peroxide led to an increase in the intracellular pools of putrescine and potassium, promotion of cellular energy status, and the restoration of DNA supercoiling to values much in excess of the prestress level. The subsequent stabilization of the intracellular putrescine pool was accompanied by a release of this polyamine from the cell. Based on these results and those available in the literature, a mechanism of E. coli adaptation to oxidative stress is suggested that assigns roles to putrescine, potassium, and cellular energy status. PMID- 10368801 TI - [Cross-action of extracellular stress adaptation factors in microorganisms]. AB - The capacity of microorganisms from different taxa to adapt to stress conditions with the help of extracellular factors exhibiting similar mechanisms of action was demonstrated. The action of adaptation factors synthesized by the enteric bacterium Escherichia coli, the soil bacterium Bacillus subtilis, and the yeast Candida utilis was characterized in biological tests. It was demonstrated that the factor of accelerated adaptation to new media (FAANM) and the growth-rate reducing factor (factor X(II)), which decreases the rate of exponential culture growth, were synthesized by all microorganisms tested. Antilysin (factor X(I)), which accelerates cell adaptation to N-ethylmaleimide, was not produced by C. utilis. PMID- 10368802 TI - [Effect of the RNAase from Bacillus intermedius on growth and physiological characteristics of Escherichia coli]. AB - The effect of the RNase from Bacillus intermedius on the growth of Escherichia coli was investigated. RNase added to growth medium enhanced the synthesis of DNA, RNA, and protein and stimulated cell division; the degree of stimulation depended on the enzyme concentration. A necessary condition for stimulation was the adsorption of the enzyme on the cell surface and its interaction with the cytoplasmic membrane, as demonstrated immunocytochemically. The adsorption of the enzyme was accompanied by a 43% decrease in the surface charge density. Other effects of RNase involved a 25% increase in the growth rate, a 38% biomass gain, and generation time shortening by 10 min. The stimulation of bacterial growth correlated with the stimulation of cellular respiration rate. PMID- 10368803 TI - [Interferon beta-1b. New therapy for patients with secondary progressive multiple sclerosis]. PMID- 10368804 TI - In search of hypotheses. PMID- 10368805 TI - Evaluation of pancreatic proteolytic enzyme treatment of adenocarcinoma of the pancreas, with nutrition and detoxification support. AB - Historically, large doses of proteolytic enzymes, along with diet, nutritional supplements, and "detoxification" procedures, have been used in alternative therapies to treat all forms of cancer, without formal clinical studies to support their use. A 2-year, unblinded, 1-treatment arm, 10-patient, pilot prospective case study was used to assess survival in patients suffering inoperable stage II-IV pancreatic adenocarcinoma treated with large doses of orally ingested pancreatic enzymes, nutritional supplements, "detoxification" procedures, and an organic diet. From January 1993 to April 1996 in the authors' private practice, 10 patients with inoperable, biopsy-proven pancreatic adenocarcinoma were entered into the trial. After one patient dropped out, an 11th patient was added to the study (however, all 11 are considered in the data tabulation). Patients followed the treatment at home, under the supervision of the authors. As of 12 January 1999, of 11 patients entered into the study, 9 (81%) survived one year, 5 (45%) survived two years, and at this time, 4 have survived three years. Two patients are alive and doing well: one at three years and the other at four years. These results are far above the 25% survival at one year and 10% survival at two years for all stages of pancreatic adenocarcinoma reported in the National Cancer Data Base from 1995. This pilot study suggests that an aggressive nutritional therapy with large doses of pancreatic enzymes led to significantly increased survival over what would normally be expected for patients with inoperable pancreatic adenocarcinoma. PMID- 10368806 TI - p53-independent apoptosis induced by genistein in lung cancer cells. AB - Lung cancer is the leading cause of cancer-related deaths in the world, with increasing incidence in many developed countries. Epidemiological data suggest that consumption of soy products may be associated with a decreased risk of cancer. Despite the association of nutrition and cancer, the molecular mechanisms by which the active metabolite in the soy diet, genistein, exerts its biological response have not been studied. We previously showed that genistein can inhibit the growth of H460 non-small-cell lung cancer (NSCLC) cells in vitro. To explore the molecular mechanisms by which genistein inhibits the growth of NSCLC cells, we investigated cell growth inhibition, modulation in gene expression, and induction of apoptosis by genistein in H460 cells, which harbor wild-type p53, and H322 cells, which possess mutated p53. Genistein was found to inhibit H460 and H322 cell growth in a dose-dependent manner. Staining with 4,6-diamidino-2 phenylindole, poly(ADP-ribose) polymerase cleavage, and flow cytometric apoptosis analysis were used to investigate apoptotic cell death, and the results show that 30 microM genistein causes cell death via a typical apoptotic pathway. Western blot analysis demonstrated upregulations of p21WAF1 and Bax by genistein in wild type and mutant p53 cell lines. Furthermore, cells treated with genistein showed an increased expression of endogenous wild-type p53, while the level of the mutant p53 protein remained unchanged. From these results, we conclude that genistein induces apoptosis in NSCLC cells through a p53-independent pathway and, thus, may act as an anticancer agent. PMID- 10368808 TI - Dietary intake and sources of isoflavones among Japanese. AB - We examined the dietary intake and sources of isoflavones (daidzein and genistein) among Japanese subjects based on dietary records (DRs). The subjects comprised two groups: 1,232 who completed one-day DRs (Group 1) and 88 men and women who kept four four-day (16-day) DRs. For quantitative data on the level of daidzein and genistein in soy foods, we extensively reviewed the literature, particularly for Japanese soy foods, and adopted the median value for each food. The median intake of daidzein was 12.1 and 9.5 mg/day among Groups 1 and 2, respectively, while the corresponding values for genistein were 19.6 and 14.9 mg/day. The top four foods (tofu, miso, natto, and fried tofu) covered about 90% of the population intake of daidzein and genistein. It did not seem feasible to estimate one's intake of isoflavones by using dietary recording/recall in epidemiological studies, since the day-to-day variation in intake was too large, the within-person coefficient of variation being 89.1% for daidzein and genistein. Therefore, we should use other methods, such as food-frequency questionnaires, focusing on the four major sources of isoflavones, to assess individual isoflavone intake. PMID- 10368807 TI - Effect of dietary conjugated linoleic acid on phorbol ester-induced PGE2 production and hyperplasia in mouse epidermis. AB - Conjugated linoleic acid (CLA) is a chemoprotective fatty acid that inhibits phorbol ester-induced skin tumor promotion in mice. The goal of the present study was to determine potential chemoprotective mechanisms through which CLA may be acting. Mice were fed diets containing 0.0%, 0.5%, 1.0%, or 1.5% CLA (by wt) for six weeks. The epidermis was evaluated for fatty acid composition, vascular permeability, prostaglandin E2 (PGE2) production, hyperplasia, ornithine decarboxylase activity, and c-myc mRNA accumulation. Fatty acid analysis of mouse epidermis demonstrated a dose-dependent increase of CLA incorporation into phospholipids and neutral lipids. In mice topically treated with the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA), dietary CLA (1.5%) significantly (p < 0.05) reduced PGE2 synthesis (2-fold). Additionally, CLA lowered accumulation of c-myc mRNA, a gene commonly associated with regulating cell cycle components involved in cellular proliferation, although this trend was not significant. Vascular permeability was unaffected by dietary CLA. These data suggest that dietary CLA modulates TPA-induced tumor promotion through a mechanism involving PGE2 production; however, dietary CLA had a moderate effect on c-myc mRNA levels and little effect on TPA-induced hyperplasia and ornithine decarboxylase activity. PMID- 10368809 TI - Effects of oral administration of tea, decaffeinated tea, and caffeine on the formation and growth of tumors in high-risk SKH-1 mice previously treated with ultraviolet B light. AB - Treatment of SKH-1 mice with ultraviolet B light (UV-B, 30 mJ/cm2) twice a week for 22-23 weeks resulted in tumor-free animals with a high risk of developing malignant and nonmalignant tumors during the next several months in the absence of further UV-B treatment (high-risk mice). In three separate experiments, oral administration of green tea or black tea (4-6 mg tea solids/ml) as the sole source of drinking fluid for 18-23 weeks to these high-risk mice inhibited the formation and decreased the size of nonmalignant squamous cell papillomas and keratoacanthomas as well as the formation and size of malignant squamous cell carcinomas. In one experiment all these inhibitory effects of tea were statistically significant, whereas in the two other experiments many but not all of the inhibitory effects of tea were statistically significant. The decaffeinated teas were inactive or less effective inhibitors of tumor formation than the regular teas, and adding caffeine back to the decaffeinated teas restored biological activity. Oral administration of caffeine alone (0.44 mg/ml) as the sole source of drinking fluid for 18-23 weeks inhibited the formation of nonmalignant and malignant tumors, and this treatment also decreased tumor size in these high-risk mice. PMID- 10368811 TI - Serum and tissue lycopene and biomarkers of oxidation in prostate cancer patients: a case-control study. AB - Dietary intake of tomatoes and tomato products containing lycopene, an antioxidant carotenoid, has been shown in recent studies to reduce the risk of cancer. This study was conducted to investigate the serum and prostate tissue lycopene and other major carotenoid concentrations in cancer patients and their controls. Serum lipid and protein oxidation was also measured. Twelve prostate cancer patients and 12 age-matched subjects were used in the study. Significantly lower serum and tissue lycopene levels (44%, p = 0.04; 78%, p = 0.050, respectively) were observed in the cancer patients than in their controls. Serum and tissue beta-carotene and other major carotenoids did not differ between the two groups (p = 0.395 and p = 0.280, respectively). Although there was no difference (p = 0.760) in serum lipid peroxidation between cancer patients and their controls (7.09 +/- 0.74 and 6.81 +/- 0.56 mumol/l, respectively), serum protein thiol levels were significantly lower among the cancer patients (p = 0.026). This study demonstrates that the status of lycopene but not other carotenoids in prostate cancer patients is different from controls. The role of dietary lycopene in preventing oxidative damage of biomolecules and thereby reducing the risk of prostate cancer needs to be evaluated in future studies. PMID- 10368810 TI - Flaxseed and lignans increase cecal beta-glucuronidase activity in rats. AB - Flaxseed has been shown in previous studies to decrease some early markers of colon cancer risk in part because of its lignans. This study determined whether the intake of flaxseed and lignans is related to the activity of bacterial beta glucuronidase, an enzyme suggested to increase colon cancer risk. Seven groups of six female rats each were fed, for four weeks, a basal high-fat (20%) diet (BD), BD supplemented with 2.5%, 5.0%, or 10.0% flaxseed, or BD with daily gavage of 0.75, 1.5, or 3.0 mg of secoisolariciresinol diglycoside (SDG), the major mammalian lignan precursor. The specific and total activities of beta glucuronidase in the cecum were significantly related to the levels of flaxseed (r = 0.539, p < 0.008 and r = 0.599, p < 0.002, respectively) and SDG (r = 0.567, p < 0.007 and r = 0.435, p < 0.04, respectively). The urinary mammalian lignan excretion also increased with increasing flaxseed or SDG levels and thus was significantly related to the specific activity (r = 0.38, p < 0.017) and total activity (r = 0.429, p < 0.007) of beta-glucuronidase. Because flaxseed and lignans are colon cancer protective, it is concluded that, in contrast to other studies, beta-glucuronidase activity may play a beneficial role in their presence by increasing mammalian lignan absorption and enterohepatic circulation. PMID- 10368812 TI - Effects of the Bowman-Birk inhibitor on clonogenic survival and cisplatin- or radiation-induced cytotoxicity in human breast, cervical, and head and neck cancer cells. AB - Bowman-Birk inhibitor (BBI) is a soybean-derived anticarcinogenic protease inhibitor previously shown to potentiate cisplatin-induced cytoxicity in human lung and ovarian cancer cells. To further assess the potential of BBI as a sensitizing agent for cancer radiotherapy and chemotherapy, we evaluated the effects of BBI and a soybean concentrate enriched in BBI known as BBI concentrate (BBIC) on clonogenic survival and radiation- or cisplatin-induced cell killing in MCF7 human breast carcinoma cells, SCC61 and SQ20B human head and neck carcinoma cells, HeLa, HeLa-R1, and HeLa-R3 human cervical carcinoma cells, MCF10 nontumorigenic human epithelial cells, HTori-3 nontumorigenic human thyroid epithelial cells, and C3H10T1/2 mouse fibroblast cells. BBI and BBIC significantly suppressed the clonogenic survival of MCF7 and SCC61 cells. BBIC also suppressed the survival of SQ20B cells and enhanced radiation-induced cell killing in SCC61 and SQ20B cells and cisplatin-induced cell killing in HeLa, HeLa R1, and HeLa-R3 cells. In contrast, BBI and/or BBIC did not enhance radiation induced cell killing in MCF10 cells or cisplatin-induced cell killing in C3H10T1/2 cells. BBI did not significantly affect the survival of SQ20B cells or enhance radiation-induced cell killing in SCC61 and SQ20B cells. The clonogenic survivals of MCF10 and C3H10T1/2 cells were not adversely affected by treatment with BBI or BBIC. The clonogenic survival of HTori-3 cells was only moderately suppressed by treatment with BBIC at > or = 80 micrograms/ml. These results suggest that BBIC could be a useful agent for the potentiation of radiation- and cisplatin-mediated cancer treatment without significant adverse effects on surrounding normal tissues. PMID- 10368813 TI - Bowman-Birk inhibitor suppresses production of superoxide anion radicals in differentiated HL-60 cells. AB - The Bowman-Birk protease inhibitor (BBI) is a soybean-derived protease inhibitor with anticarcinogenic and anti-inflammatory properties. BBI has previously been shown to suppress the release of superoxide anion radicals from purified polymorphonuclear leukocytes. In the present study we evaluated the effect of BBI on the production of superoxide anion radicals in differentiated HL-60 cells. HL 60 cells are human lymphocytic cells that acquire neutrophil-like characteristics when treated with dimethyl sulfoxide or tetradecanoyl phorbol acetate. Superoxide anion radical production by differentiated HL-60 cells was measured in the presence of various concentrations of BBI or BBI concentrate, a soybean extract containing high levels of BBI. BBI was observed to suppress superoxide anion radical production by differentiated HL-60 cells in a dose-dependent manner. Extracts of differentiated HL-60 cells were also observed to produce superoxide anion radicals, but this activity was not affected by the presence of BBI. These results suggest that BBI inhibits superoxide anion radical generation in HL-60 cells but does not act as a simple free radical scavenger. PMID- 10368814 TI - Modulation of cytochrome P-450 and glutathione S-transferase isoform expression in vivo by intact and degraded indolyl glucosinolates. AB - Various dietary substances modulate the xenobiotic metabolism and may thereby protect against toxicity and carcinogenicity of food toxins. The effects of pure indolyl glucosinolates, which are present in cruciferous vegetables, on induction of specific cytochrome P-450 (CYP) and glutathione S-transferase (GST) isoforms have not been studied previously. In the present study, glucobrassicin (GB) and neoglucobrassicin (NeoGB) were purified from broccoli by use of a single-column method. Furthermore, a mixture containing 48% GB, 36% NeoGB, and 16% 4 methoxyglucobrassicin was obtained. The modulatory effects of the pure GB, NeoGB, and the mixture on activities and levels of hepatic CYP 1A, 2B1/2, and 2E1 and alpha- and mu-GST isoforms were investigated in male Wistar rats. The indolyl mixture was the most powerful and NeoGB the weakest inducer of microsomal hepatic CYP 1A1 protein and 7-ethoxyresorufin O-deethylase activity. Furthermore, intact indolyl glucosinolates were more powerful inducers than the in vitro myrosinase degraded indolyl glucosinolates. The hepatic 7-methoxyresorufin O-deethylase activities, but not CYP 1A2 protein, were induced by pure GB, whereas the mixture and NeoGB showed only minor effects. Neither CYP 2B1/2 nor 2E1 was induced by the indolyl glucosinolates. None of the hepatic GST subunits analyzed, rGST A1/2, A3, or M3, was induced significantly by the purified indolyl glucosinolates. PMID- 10368815 TI - Effect of flaxseed consumption on urinary estrogen metabolites in postmenopausal women. AB - Flaxseed, the richest known source of plant lignans, has been shown to have chemoprotective effects in animal and cell studies. Some of its effects may be mediated through its influence on endogenous hormone production and metabolism. Two competing pathways in estrogen metabolism involve production of the 2 hydroxylated and 16 alpha-hydroxylated metabolites. Because of the proposed differences in biological activities of these metabolites, the balance of the two pathways has been used as a biomarker for breast cancer risk. We examined the effects of flaxseed consumption on urinary estrogen metabolite excretion in postmenopausal women. Twenty-eight postmenopausal women were studied for three seven-week feeding periods in a randomized crossover design. During the feeding periods, subjects consumed their usual diets plus ground flaxseed (0, 5, or 10 g/day). Urinary excretion of the estrogen metabolites 2-hydroxyestrogen (2 OHEstrogen) and 16 alpha-hydroxyestrone (16 alpha-OHE1) as well as their ratio, 2/16 alpha-OHE1, was measured by enzyme immunoassay. Flaxseed supplementation significantly increased urinary 2-OHEstrogen excretion (p < 0.0005) and the urinary 2/16 alpha-OHE1 ratio (p < 0.05) in a linear, dose-response fashion. There were no significant differences in urinary 16 alpha-OHE1 excretion. These results suggest that flaxseed may have chemoprotective effects in postmenopausal women. PMID- 10368816 TI - Population-attributable risk for colon cancer in Italy. AB - Risk factors for colon cancer have essentially been considered in terms of relative risks. From a public health viewpoint, however, their impact depends not only on the strength of the association, but also on the distribution of exposures in the population. Thus we used data from a case-control study conducted in Italy between 1992 and 1996 to estimate the population-attributable risks (PARs) for colon cancer in relation to educational level, physical activity, energy and vegetable intake, eating frequency, and family history of colorectal cancer. Cases were 1,225 incident, histologically confirmed colon cancer patients admitted to the major teaching and general hospitals in six Italian areas; controls were 4,154 subjects with no history of cancer, admitted to hospitals in the same catchment areas for acute, nonneoplastic diseases. By use of the distribution of the risk factors in the cases and the multivariate relative risk estimates, PARs were computed, i.e., the proportion of colon cancer that would have been avoided if all subjects were moved to the lowest exposure level. The PARs were 12% for high education, 14% for low physical activity, 14% for high energy intake, 22% for low vegetable intake, 7% for high eating frequency, and 8% for a family history of colorectal cancer. These factors together accounted for 56% of colon cancer cases. PARs were similar across age strata. Men had higher PARs for education, physical activity, and their combination, but lower PARs for energy, eating frequency, vegetable intake, and their combination than women. The percentage of colon cancers attributable to all factors considered together was 50% in men and 67% in women. Even if the PAR estimates were based on several arbitrary assumptions on the exposure distribution for various risk factors, available knowledge could, in principle, explain > 50% of cases in this Italian population, thus indicating and quantifying the theoretical scope for prevention. PMID- 10368817 TI - Lipolytic activity of anemia-inducing substance from tumor-bearing rabbits. AB - Anemia-inducing substance (AIS) is a protein of approximately 50,000 molecular weight secreted by malignant tumor tissue that depresses erythrocyte and immuno competent cell functions; in this study, its biological effects on adipocytes were examined. Changes in body weight, total body fat, and food intake were investigated in rabbits after VX2 carcinoma transplantation, and the results showed reductions of 11%, 24%, and 30%, respectively, at 40 days after transplantation compared with baseline values (before transplantation). The values were even more markedly reduced 70 days after transplantation. When cyclic plasma perfusion (2 times/wk) was started at 40 days after transplantation, the values at 70 days after transplantation (30 days after beginning plasma perfusion) recovered to 91%, 84%, and 87%, respectively, of the baseline values. AIS fractions were isolated from rabbit plasma by using a phenyl-Sepharose column before transplantation, 40 and 70 days after transplantation, and 30 days after start of plasma perfusion, and AIS activity and lipolytic activity were measured. The results showed enhancement of AIS activity and lipolytic activity as the tumors grew. Lipolytic activity also returned to baseline value as AIS was removed by adsorption by plasma perfusion, and there was a high correlation between lipolytic activity and AIS kinetics. These results strongly suggest that AIS might be one of the substances involved in the enhanced lipolytic activity in advanced tumor-bearing subjects. PMID- 10368818 TI - Dietary lutein but not astaxanthin or beta-carotene increases pim-1 gene expression in murine lymphocytes. AB - This study investigates the effect of dietary carotenoids on pim-1 gene expression in mouse splenocytes. Female BALB/c mice were fed 0%, 0.02%, or 0.4% astaxanthin, beta-carotene, and lutein for two weeks. Plasma and liver were obtained for the analysis of carotenoids. Splenocytes were isolated and cultured in the presence of concanavalin A, and the level of pim-1 mRNA was determined by Northern blot analysis. None of the carotenoids were detectable in the plasma and liver of unsupplemented mice. In plasma the concentration of astaxanthin (4.9 54.7 mumol/l) was dramatically higher than that of lutein (1.4-2.0 mumol/l) and beta-carotene (0.1-0.7 mumol/l). Carotenoid uptake by the spleen but not the liver reflected that observed in plasma. In mice fed 0.4% of each carotenoid, the absolute concentration of the carotenoid in the liver was highest for astaxanthin (24 nmol/g) followed by beta-carotene (7.5 nmol/g) and lutein (1.58 nmol/g). Mice fed lutein showed a dose-related increase in pim-1 mRNA expression. The steady state level of pim-1 mRNA in mice fed 0.4% lutein was sixfold higher than in mice fed 0.02% lutein. In contrast, dietary astaxanthin and beta-carotene did not affect pim-1 expression. Therefore, an increase in pim-1 mRNA was observed in splenocytes stimulated with concanavalin A in lutein-fed mice. This appears to be a unique effect of lutein and may be associated with its antitumor activity observed in vivo. PMID- 10368819 TI - Utility of produce ratios to track fruit and vegetable consumption in a rural community, church-based 5 A Day intervention project. AB - Previous research suggests that grocery store characteristics may be useful in evaluating community-based dietary interventions. We undertook a study to determine whether produce ratios (ratios of produce sales to total grocery sales) were a useful indicator of fruit and vegetable (F & V) consumption in a church based, community intervention trial that promoted 5 A Day guidelines within 10 rural counties of North Carolina. Produce ratios were collected from stores identified by participants in the Black Churches United for Better Health Project. Baseline and study period data for 21 stores in intervention counties and 18 stores in nonintervention counties were compared using repeated-measures analysis of variance. Produce ratios were significantly associated with seasonality (p < 0.0001), but no differences were seen between the two groups of stores. These findings do not support data from individual telephone surveys, which showed significant differences in F & V consumption between participants in the two groups. Our inability to detect differences at the store level may have been due to 1) the incapacity of produce ratios to capture F & V purchases that were juice, frozen, or canned products; 2) shifts in procuring F & Vs from grocery stores to other sources (i.e., gleaning and produce cooperatives); 3) the modest proportion of shoppers that received the full intervention dose; and 4) a general lack of power to detect differences at the store level. Therefore, although produce ratios did not serve as a valid measure for this project, if their limitations are recognized and compensated for, they may have applicability for future investigations that monitor F & V consumption. PMID- 10368820 TI - [The translation regulation of the synthesis of proteins responsible for dorsoventral differentiation of clawed toad embryos]. AB - We studied the distribution of mRNAs involved in dorsoventral differentiation of Xenopus laevis embryos (Xbra, chordin, Xnot, Xvent1, Xvent2, and Xwnt11) among polyribosomes (the translated form of mRNA) and informosomes (the nontranslated form of mRNA). It was shown using molecular hybridization that all of the studied templates are in the informosomes until the midgastrula stage, thus suggesting regulation at the level of translation. At the midgastrula stage (stage 11), translation begins on mRNAs of chordin, Xnot2, Xvent1, and Xbra, although mRNAs of Xvent1 and Xbra are partially located in the informosomes. The matrices of Xwnt11 are localized predominantly in the informosomes at the midgastrula stage, while those of Xvent2 are not seen in the polyribosomes until the end of gastrulation. We propose that the fate of different mRNAs is determined by different mechanisms of r-translational regulation in different cell lineages. It cannot be excluded that the Xvent2 transcript is not involved in translation and fulfills its morphogenetic functions in the form of RNA or RNP. PMID- 10368822 TI - [The levels and nature of translation in mouse preimplantation embryos with repressed cytokinesis]. AB - The translational activity of embryos cultured up to the 8-cell stage in cytochalasin D-supplemented medium was studied. These embryos remained unicellular during the entire preimplantation period. Blastocoel formation and hatching began in the cytochalasin-treated embryos at the same time as in the control. We have studied the overall translation rates of "one-cell embryos" and estimated the relative translational differences for individual polypeptides compared to the control embryos. Up to the early blastocyst stage, the translation levels in these embryos, measured by 35S-methionine incorporation, were about two times lower than in the control. At later stages, the differences were fivefold. The ppm values of approximately one-third of all individual spots on two-dimensional electrophoregrams differed three times or more between the control and cytochalasin-treated embryos. These results suggest that the first morphogenetic events in mammalian ontogenesis may be controlled autonomously and the timing of morphogenetic transitions is controlled by acquisition of a definite set of stage-specific factors which serves as the signal for subsequent development. PMID- 10368821 TI - [The effect of hyperbaric oxygenation on the antioxidant status of Xenopus laevis after its preliminary adaptation to oxygen]. AB - We studied the level of lipid peroxidation and the activity of antioxidant enzymes (superoxide dismutase and catalase) in various tissues of adult Xenopus laevis after an initial exposure to hyperbaric oxygenation at the developmental stage 38. We have found that irrespective to the mode of treatment, the level of lipid peroxidation and activity of antioxidant enzymes in the brain, lungs, and blood of these animals were higher as compared to control animals. We demonstrate that, after the exposure of adult animals to hyperoxia, if they were earlier subjected to hyperbaric oxygenation (0.2 MPa) at stage 38, there was no intensification of lipid peroxidation or changes in the activity of superoxide dismutase and catalase. In adult animals initially subjected to hyperbaric oxygenation at the same stage of development but at the pressure--0.7 MPa, the second exposure to hyperoxia led to a drastic intensification of lipid peroxidation in the brain; in some animals, an increased level of lipid peroxidation products in the lungs was observed. PMID- 10368823 TI - [The factors affecting the level of ovarian hormone receptors in breast tumors in women]. AB - We studied the level of estradiol and progesterone receptors in breast tumors of female patients. We have shown that the level of these receptors in tumors depends on whether the tumour is present in the left or right breast, the age of the patient, and the season of the year. PMID- 10368824 TI - Inhibitors of aspartic proteases in human diseases: molecular modeling comes of age. AB - The family of aspartic proteases such as cathepsin D, gastricin, pepsin, renin, HIV protease and others have been the subject of molecular modeling in the field of drug design in the last years. The first aspartic protease inhibitor was reported thirty years ago as a renin inhibitor. The success of HIV protease inhibitors in preventing progression to AIDS was based on the transition state analogs of renin inhibitors. Taking these three decades into consideration, an astonishing variety of chemical classes, in vitro and in vivo activities and species specificities of inhibitors of aspartic proteases have been reported. Especially inhibitors of renin, HIV protease and secreted aspartic protease of Candida albicans are covered. PMID- 10368825 TI - New benzoxazine and benzothiazine derivatives--structure-activity relations in guinea-pig heart and smooth muscle preparations. AB - New benzoxazine and benzothiazine derivatives which differ in their side chains on the nitrogen atom of the benzoxazine or benzothiazine ring were investigated regarding structure-activity relations and compared with calcium antagonistic drugs. The isometric contraction force was measured in guinea-pig papillary muscles, aortic strips and terminal ilea. Chronotropic activity was studied in right atria of guinea pigs. The benzoxazine derivative with a dimethoxyphenylethyl-N-methylaminoethylcarboxamide side chain (MS 84) had the most potent negative inotropic effect on papillary muscles and the most potent relaxing effect on terminal ilea. The benzothiazine derivative with a methylpiperazinylcarbonyl side chain (MS 63) was less effective. Benzoxazine derivatives with a methylpiperazinylcarbonyl (MS 64) or a N dimethylaminoethylcarboxamide side chain (MS 66) and the benzothiazine derivative (MS 65) with an analog side chain like MS 66 only had a weak effect. We conclude that the oxygen atom in the heterocyclic ring and the lipophilic side chain on the nitrogen atom, which is almost identical with the calcium antagonistic drug KT-362 is responsible for the most potent action. PMID- 10368827 TI - Benzimidazole condensed ring systems. XII. Synthesis and anticancer evaluation of certain pyrido[1,2-a]benzimidazole derivatives. AB - Previously, we have evaluated several pyrido[1,2-a]benzimidazoles (PBIs) as potential antineoplastic agents. Among them, NSC 649900 and NSC 682011 revealed good antineoplastic activity against some cell lines of clinically isolated human tumors. For further structure-activity relationship (SAR) studies we report here the synthesis and antineoplastic evaluation of related series of PBIs with similar haloarylamino (13-18, 23-28), haloarylaminomethylene (29-34) and haloarylazo (35-38) moieties at position 1 or 2. Some of these derivatives revealed notable activity against some tumor cell lines; the highest activity was recorded for the p-fluorophenylamino-3-phenyl-PBI (23, NSC 699944) and its p chlorophenyl analog (24, NSC 699948). These compounds were selected by the NCI for further testing in a new in vivo anticancer hollow fiber assay. PMID- 10368826 TI - Fused 1,2-dithioles, V: Carbenoid anions as intermediates in reactions of pyrrothines and their heteroanalogues. AB - Pyrrothines like thiolutine and other bicyclic 1,2-dithioles of type 1 when unsubstituted in 3-position are marked by their CH acidity. In the presence of weak bases such as triethylamine the pyrrothine 4 degraded via its anion to a thioketene trapped as the 1,3-dithietane 5. The carbenoid anions of several compounds 1 reacted with elemental sulphur forming enthiolates whose alkylation led to the corresponding thioethers or, in the case of the thiolactam 9, to the bicyclic trithiones 10 and 11. In the same manner selenides can be obtained via intermediate selenolate ions. Introduction of an aryl- or heteroarylmercapto group into compounds 1 was achieved directly by reaction of the corresponding anions with suitable disulphides. Though there may be structural limitations, this sulphurization reaction can be extended to 3-unsubstituted trithiones. The new compounds exhibited significant activity against Mycobacterium tuberculosis in the primary screening. PMID- 10368828 TI - Synthesis of heterobicyclic nitrogen systems bearing the 1,2,4-triazine moiety as anti-HIV and anticancer drugs: Part I. AB - Some new heterobicyclic nitrogen systems bearing the 1,2,4-triazine moiety (3-22) have been achieved by treatment of 3-amino-5,6-disubstituted-1,2,4-triazines 2a-h with some cyclic and acyclic oxygen compounds followed by heterocyclization. Structures of the products have been deduced by elemental analysis and spectral data. Significant anti-HIV and anticancer activities were observed in vitro for some members of the series, where compounds 5 and 18 showed a moderate activity. PMID- 10368829 TI - Synthesis and immunological activity of new 5-amino-3-methyl 4-amido and 4 ureilene isoxazole derivatives. AB - 5-Amino-3-methylisoxazole-4-carboxylic acid amides and ureilenes have been synthesized from 5-amino-3-methylisoxazole-4-carbonyl azide. The compounds were investigated for potential immunotropic activity in several immunological tests. The most interesting suppressory activities in the humoral and cellular immune response were compared to activities of analogous compounds previously described as immunostimulatory. PMID- 10368830 TI - Comparative in vitro study of different chitosan-complexing agent conjugates. AB - In this study we analysed the bioadhesive properties and the enzyme inhibitory effects of different chitosan-complexing agent conjugates. Etylenediaminetetraacetic acid (EDTA) and diethylenetriaminepentaacetic acid (DTPA), respectively, were covalently attached to chitosan by the formation of amide bonds between the primary amino group of the polymer and the carboxylic acid groups of the complexing agents. Whereas almost each primary amino groups of chitosan could be modified by EDTA, DTPA was bound to only 63.8 +/- 5.8% (n = 3; +/- SD) of the amino groups of chitosan. The remaining primary amino groups of the chitosan-DTPA conjugate lead to strongly reduced adhesive properties, with a maximum detachment force of 3.0 +/- 1.3 mN in contrast to the chitosan-EDTA conjugate with 81.7 +/- 9.9 mN in the tensil studies described here (n = 4; +/- SD). However, both polymer conjugates displayed an inhibitory effect towards the zinc-dependent proteases carboxypeptidase A (EC 3.4.17.1) and aminopeptidase N (EC 3.4.11.2). The results of this comparative study should provide substantial knowledge for the development of bioadhesive polymers as auxiliary agents for the peroral administration of peptide and protein drugs. PMID- 10368831 TI - Preformulation stability screening of ivermectin with non-ionic emulsion excipients. AB - As an important and complementary step during a preformulation study differential scanning calorimetry (DSC) and high-pressure liquid chromatography (HPLC) were used to determine the compatibility between ivermectin and commonly used excipients for preparing non-ionic emulsions. Ivermectin was found to exhibit interactions with 21 excipients, while it was compatible with 25 excipients. HPLC showed a significant decrease in drug amount +20%, when substances interacted with invermectin. PMID- 10368832 TI - Magnetophoresis: an approach to enhance transdermal drug diffusion. AB - Magnetophoresis is a novel approach in enhancing drug delivery across biological barriers. Benzoic acid, a diamagnetic substance, was selected as the drug candidate. The drug diffusion across rat abdominal skin was enhanced due to the influence of the magnetic field. The experiment with alternating on-off-fields in the same diffusion set-up confirmed that the difference in flux between passive and magnetic diffusion is not due to any variation in the experimental condition or membrane properties. The influence of magnetic field strength on diffusion flux was determined and was found to increase with increasing applied field strength. PMID- 10368833 TI - Nonspecific membrane effects of CH-103: hydrophobicity, surface activity and membrane fluidity studies in comparison with propranolol and practolol. AB - The partition coefficient, surface activity and membrane fluidizing/disordering effects of CH-103, a beta-adrenergic receptor antagonist, were compared to those of propranolol and practolol as reference compounds. Changes in membrane fluidity were followed by measuring the steady-state fluorescence anisotropy of bull sperm cells with 1-[4-(trimethylammonium)-phenyl]-6-phenyl-1,3,5-hexatriene (TMA-DPH) as a fluorescence probe. The octanol/buffer (pH 7.0) partition coefficients for CH-103, propranolol and practolol were 32.9, 5.08 and 0.013, respectively; the surface activity of the compounds decreased in the same order. CH-103 and propranolol significantly increased the fluidity of the membrane in a concentration-dependent manner, whereas practolol reduced fluidity. These physicochemical parameters correlated with the effects of these drugs on rat sarcolemmal Ca2+, Mg(2+)-ATPase, a manifestation of their nonspecific membrane activity. Our results suggest that the physicochemical properties of CH-103, similarly to those of propranolol, are the main determinants of its nonspecific membrane activity. PMID- 10368834 TI - Research designs in physiotherapy, which is best. PMID- 10368835 TI - A study of repeated lateral pinch grip in myotonic dystrophy. AB - BACKGROUND AND PURPOSE: Subjects with myotonic dystrophy present with progressive muscle weakness, myotonia and fatigue. The aim of this study was to determine whether there was a difference in response to fatiguing exercise in myotonic dystrophy individuals compared to normal subjects. If no difference was found, a similar response to a physiotherapy exercise programme as seen in normal subjects might be expected. METHOD: Ten individuals with myotonic dystrophy were compared to eight normal subjects in their response to ten repetitions of maximal lateral pinch grip efforts each five seconds in duration and separated by a ten-second rest. The root mean square (RMS) values, initial median frequency (Fmed) and slope of the median frequencies were recorded by electromyography (EMG) for the first dorsal interosseus, flexor pollicis brevis, flexor digitorum profundus and extensor digitorum communis muscles in the forearm and hand. Simultaneously, the rate of grip development, rate of grip release and work done during each grip effort were recorded by dynamometer. The RMS values for all muscles from subjects with myotonic dystrophy increased over the ten repetitions. RESULTS: The initial Fmed for all myotonic dystrophy muscles was lower than for the normal subjects. The Fmed slopes for the first and last repetition showed no significant difference to the normal subjects. Rate of grip development was no different between groups over ten repetitions. Rate of grip release was slower and work done less for the individuals with myotonic dystrophy. CONCLUSION: Results suggest the main difference between muscles affected by myotonic dystrophy and normal ones was the smaller size of muscle fibres. The increase in rate of grip release that was found is supportive of the 'warm-up' phenomenon. This appears to indicate that muscles affected by myotonic dystrophy could benefit from standard physiotherapeutic exercise methods. PMID- 10368836 TI - Inter-segmental co-ordination in sit-to-stand: an age cross-sectional study. AB - BACKGROUND AND PURPOSE: Being able to sit-to-stand (STS) effectively is an important functional skill, but there is little information available on the changes that occur with growth and maturity. This study aimed to investigate the inter-segmental co-ordination of STS in three different age groups (12-18 months, 4-5 years and 9-10 years). METHOD: The children studied wore reflective markers and were videotaped as they stood up from a height-adjustable seat that straddled a forceplate. Segmental kinematics and vertical ground reaction force were determined from the co-ordinate and forceplate data. RESULTS: Even at the earliest developmental stage the children had mastered the basic inter-segmental pattern observed in adults. The youngest children, however, were not able to end the movement in quiet standing; rather they raised up on their toes or took a step forward. Performance varied both within and between subjects. Although there was a similarity in the motor pattern used by the younger subjects to that of the older subjects, developmental trends were evident on the videotapes and on examination of the kinematic and kinetic variables. Movement time, amplitude and peak angular velocity of trunk flexion increased with age. Whereas the children in the older age groups displayed a pattern of vertical ground reaction force similar to that reported for adults, the youngest children tended to reach peak force gradually, often with fluctuations. Although there were characteristic trajectories in the phase-plane plots for each group, the overall trend was for the percentage of smooth plots representing a co-ordinated movement, to increase with age. CONCLUSIONS: Differences in inter-segmental co-ordination between the ages studied may relate to the child's ability to control horizontal momentum and to balance. PMID- 10368837 TI - Inappropriate sexual behaviours of patients towards practising physiotherapists: a study using qualitative methods. AB - BACKGROUND AND PURPOSE: Recent research recognizes the occurrence of inappropriate sexual behaviour (ISB) by patients towards health professionals. The objective of this study was to explore in-depth the clinical context and effect of incidents of ISB towards practising physiotherapists. METHOD: In-depth interviews were conducted with a sub-sample of nine physiotherapists who were part of a larger survey on ISB. Quantitative analyses of the survey responses are reported elsewhere. Interview participants were asked to describe an incident of ISB by a patient that was either perceived to be the worst or was the most recent. They were asked questions on a variety of themes, such as their relationship with the patient prior to incident, the effects of the incident, the strategies used to deal with the incident, and changes in practice as a result of the incident. RESULTS: All interview participants reported encountering some level of ISB from patients. Although the overall frequency of these behaviours was relatively low, the range of behaviours was diverse. Regardless of the perceived severity of the incident, only four participants labelled their experience as 'sexual harassment'. Many reported negative effects on work performance. Participants mainly used physical measures to prevent further incidents, rather than confronting the perpetrator or reporting the incident. CONCLUSIONS: The findings are discussed in the context of theory pertaining to boundaries and issues of transference and counter-transference. This emphasized the need for effective communication skills training of both undergraduate and graduate physiotherapists in the prevention and management of ISB from patients. PMID- 10368838 TI - Performance in functional balance tests during menopausal hormone replacement: a double-blind placebo-controlled study. AB - BACKGROUND AND PURPOSE: Recent studies have indicated that oestrogens might have an effect on postural control. The purpose of this study was to investigate the short-term effect of hormone replacement therapy (HRT) on postural control in menopausal women and to analyse the correlation between sway velocity measures and results of functional balance tests. SUBJECTS: 100 menopausal women who were randomized to receive either HRT or placebo for three months were included in the study. METHOD: The balance function was measured with nine different static and dynamic functional balance tests. The sway velocities were measured on a computerized force platform. RESULTS: No significant differences were found between the two groups in the results of the functional balance tests after the three-month placebo-controlled period or in the changes over time. However, some significant improvements occurred within both groups over this three-month period. The correlations between different sway velocities and the results of the functional balance tests were all very low (r < 0.35). CONCLUSION: It can not be concluded that HRT had a positive effect on the performance in the functional balance tests, as some improvement occurred in both groups. The low correlations indicate that the sway velocity and functional balance tests measure different aspects of balance function. PMID- 10368839 TI - The effects of knee extensor and flexor muscle training on the timed-up-and-go test in individuals with rheumatoid arthritis. AB - BACKGROUND AND PURPOSE: Rheumatoid arthritis frequently results in functional impairment. This study investigated the effect of a specific exercise regimen on function. METHOD: A randomized controlled assessor-blinded (N = 36) compared the effect of knee extensor and flexor muscle training on pain, the timed up and go (TUG) test and the Health Assessment Questionnaire in subjects with non-acute rheumatoid arthritis. RESULTS: Knee extensor and flexor muscle training increased isokinetic torques at speeds of 60 degrees/sec-1 and 120 degrees/sec-1 as measured by an isokinetic dynamometer (p = 0.02-0.003). The experimental group experienced a reduction in pain (p = 0.03), an improvement in TUG time (p = 0.01) and in function as measured by the Health Assessment Questionnaire (p = 0.04). CONCLUSIONS: Specific knee muscle training can be administered safely in people with non-acute rheumatoid arthritis, and may produce functional benefits. PMID- 10368840 TI - Management of spasticity in hereditary spastic paraplegia. PMID- 10368841 TI - ARs revisited. PMID- 10368842 TI - New estimates and projections of the population cohabiting in England and Wales. AB - This article describes the derivation of a new set of population estimates of those cohabiting in England and Wales in 1996. As well as estimating the population cohabiting by age and sex, the estimates have also been made by legal marital status. The article then discusses new projections to the year 2021, based on these estimates. PMID- 10368843 TI - Divorce and remarriage in England and Wales. AB - This short article provides a summary of the demographic consequences of divorce over the past two decades: the growth in the proportion of the population who are divorced; the increase in the age at divorce and the ageing of the divorced population; and the decline in the rates of remarriage after divorce. In addition, the extent and timing of remarriage after divorce are investigated, as well as the corresponding features of remarriage after being widowed. The differentials between men and women are presented and discussed throughout the article. PMID- 10368844 TI - 1996-based population projections by legal marital status for England and Wales. AB - This article presents the key results of the Government Actuary's Department's latest population projections by legal marital status for England and Wales. The new projections are based on the estimated population of England and Wales by marital status at mid-1996 and cover 1996-2021. PMID- 10368845 TI - Changes in fertility and family sizes in Europe. AB - This article describes recent trends in annual fertility rates, and generation measurements of average family size and family size distributions for eight selected European Countries. It compares the changing ages at which mothers are having children and the decline in higher order births, both over time and between countries: The way that variations and changes in annual rates influence generation measurements is also illustrated. Finally, the article compares prevailing annual rates with the long-term fertility assumption included in the most recent population projections for seven countries. PMID- 10368846 TI - The health and socio-economic circumstances of British lone mothers over the last two decades. AB - This article examines the trends in the socio-economic circumstances and health of lone mothers compared to couple mothers from 1979 to 1995 in Great Britain using secondary analysis of data from General Household Survey and covering 9,159 lone mothers and 51,922 couple mothers living in private households. The main measures are self perceived general health, limiting long-standing illness, poverty and working status. PMID- 10368847 TI - [Microbiological synthesis of virazole by immobilized cells]. AB - Based on the isotherms of adsorption of the cells of Xanthomonas campestris B-610 to glass and polyvinyl fibers, immobilized systems were produced, in which the cell content was sufficient for enzymatic synthesis of Virazole (1-[beta-D ribofuranosyl]-1,2,4-triazole-3-carboxamide) using adenosine as a donor of ribose (50-60% conversion of 1,2,4-triazole-3-carboxamide). The immobilized cells thus obtained retain their enzymatic activity for six months and can be used repeatedly. PMID- 10368848 TI - [Serine protease of Bacillus subtilis R]. AB - Properties of a protease preparation obtained by ethanol precipitation of a concentrate of the culture fluid of Bacillus subtilis R (1 : 4 v/v; 4 degrees C) grown under the conditions of deep cultivation were studied. The use of specific inhibitors, EDTA and phenylmethylsulfonyl fluoride, made it possible to show that the enzyme belongs to the group of serine proteases. The preparation exhibited high stability in alkaline medium and thermostability; it hydrolyzed protein substrates and retained catalytic properties in the presence of a multicomponent detergent system. The preparation is recommended for use in those branches of industry where proteolysis is required and in the production of detergents (as a biological additive). PMID- 10368849 TI - [Protective effects of agar during immobilization of strains capable to degrade aromatic compounds]. AB - A study of utilization of toxic monoaromatic compounds by microbial cells incorporated into agar matrix revealed positive effects of immobilization on the rate and completeness of biodegradation. This increase in the degrading activity is probably due to a strong protective effect of the polysaccharide carrier. The protective properties of agar are not due to sorptional and diffusional processes in the matrix. These properties were shown to be associated with the formation of a specific extracellular membrane around microcolonies. PMID- 10368850 TI - [Activity of exoglycans as sorbents of heavy metal ions]. AB - The ability of yeast exoglycans (from Cryptococcus laurentii, Cr. luteolus, and Sporobolomyces albo-rubescens) to absorb copper and lead ions has been studied. The sorption isotherms have been obtained, which indicates that the mechanism of interactions in the system polysaccharide-metal is complex. For the majority of the glycans, the absorption selectivity was considerably higher in the case of copper, as compared to lead. The observed discrepancy in the effects of pH and temperature on the absorption indicates that the bonds mediating ion absorption by glycans are of different nature. In conclusion, yeast exoglycans are efficient sorbents of copper and lead ions, and lutelan is the most active exoglycan in this respect. PMID- 10368851 TI - [Application of competitive immunoenzyme assay for analysis of group B fumonisins]. AB - A varriant of competitive ELISA, making it possible to detect fumonisin B1 (up to 0.4 ng/well) was developed, based on the use of specific polyclonal antibodies obtained by immunization of rabbits with a horseradish peroxidase conjugate of fumonisin B1. The minimum concentration of fumonisin B in water-acetonitrile extracts of corn grain, dry corn products, and feeds, detected by the ELISA version developed is 0.2 mg/kg. PMID- 10368852 TI - Clinical outcome to clozapine treatment in chronic psychiatric inpatients. AB - 1. A review of the medical records in a state psychiatric hospital was conducted to evaluate the clinical efficacy of the atypical antipsychotic, clozapine. 2. Using the Brief Psychiatric Rating Scale (BPRS), four groups of schizophrenic inpatients (n = 59) were operationally defined: Nonresponders (< 20% decrease from pre-drug baseline); Short-term Pharmacological Responders (20% decline, but not sustained); Long-term Pharmacological Responders (maintained a 20% decline) and Clinical Responders (maintained a 20% decline and achieved a BPRS < or = 36; the criterion of Kane et al. 1988). 3. There were 7 NRs, 13 STPRs, 21 LTPRs and 18 CRs 4. The STPR, LTPR and CR groups improved significantly within the first month of treatment and reached a 20% decrease in BPRS by 3 months. CRs required 5 months to attain a BPRS < or = 36. These criteria were reached at the same average doses (about 300-400 mg/day). 5. The proportion of CRs (30%) in this retrospective, naturalistic study, is remarkably close to the results of the definitive study by Kane et al. 1988. These results are also consistent with many of the controlled research studies of clozapine in hospitalized, treatment refractory psychiatric patients. PMID- 10368853 TI - Light mask 500 lux treatment for delayed sleep phase syndrome. AB - 1. Bright light exposure has been demonstrated as an effective treatment for circadian rhythm sleep disorders. Recent studies suggest that more moderate intensities of light might affect endogenous rhythms. A light mask treatment, using light applied through eyelids during sleep, was tested for Delayed Sleep Phase Syndrome. 2. The active light group (n = 5) received 500 lux light for 3 hours prior to awakening for 12 days. The placebo light group (n = 5) received 0.1 lux light with the same timing. Circadian rhythm phase was assessed from core body temperature and urinary 6-sulfatoxymelatonin measurements. The SIGH-SAD-SR mood scale was administered to assess mood. 3. There were slight trends toward a phase advance of the body temperature rhythm and a phase delay of the melatonin rhythm, and both groups reported anti-depressant benefits. However, no statistically significant effects of 500 lux light mask treatment were demonstrated compared with the placebo-light treatment. 4. More extensive studies will be required to clarify the factors of dose-response and phase-response. PMID- 10368855 TI - Depressive symptoms and schizophrenic relapses: the effect of four neuroleptic drugs. AB - 1. A prevalence of depressive symptomatology, ranging from 25% to 80% has been reported during the course of schizophrenia. 2. Depressive symptoms were assessed in 144 schizophrenic patients (DSM IV) during an acute exacerbation phase. 3. Depressive symptoms showed a prevalence ranging from 5.5% (severe clinical pictures) to 54.8 (mild clinical pictures). 4. The authors did not find a correlation between depressive symptoms per se and the presence of negative psychotic symptoms. Depression may be linked not so much to negative symptoms but to the psychotic state itself. 5. Depressive symptomatology concurrently occurred with schizophrenic relapses and improved together with the psychotic clinical picture, independently of the neuroleptic drug employed. Haloperidol, haloperidol decanoate and fluphenazine decanoate all showed a similar improvement of depressive symptoms. 6. L-sulpiride showed a trend to be most effective on depressive symptomatology in comparison to the other neuroleptics. PMID- 10368854 TI - Pharmacoclinical strategy in neuroleptic resistant schizophrenic patients treated by clozapine: clinical evolution, concentration of plasma and red blood cell clozapine and desmethylclozapine, whole blood serotonin and tryptophan. AB - 1. The aim of the study was to determine if a more rational therapeutic approach could be devised for neuroleptic resistant psychotic patients treated for months and years with clozapine. Clozapine is an atypical antipsychotic medication, but its therapeutic benefit has been limited by a high incidence of agranulocytosis and seizures. 2. The study has been performed in an open setting and included 12 patients. Some of them developed a secondary depression and were treated with fluoxetine. 3. Pharmacokinetic analysis were conducted at the same time as clinical evaluations, grading using the BPRS, the PDS, and QLS, and determinations of plasma and red blood cell clozapine and desmethylclozapine, plasma and RBC fluoxetine and norfluoxetine, whole blood serotonin and tryptophan. 4. A positive linear correlation was found only between RBC concentration and the evolution of the QLS. 5. Clozapine is efficacious both on positive and negative symptoms but its mechanism of action remains unclear. Positive symptoms disappear more quickly, sometimes followed by a post psychotic depression. Negative symptoms improve more slowly but regularly. They seem to be correlated with serotoninergic mechanisms. For whole blood 5HT, an important increase was seen about 4 weeks after Cloza administration, and then a decrease. 6. Therapeutic drug monitoring (on the same sample drawn for haematological monitoring providing) could play a useful role in the management of patients treated by clozapine: compliance, lowest dose, possible toxicity, drug interaction, lack of efficacy, relapse predictivity. PMID- 10368856 TI - A variable-number-tandem-repeat of the serotonin transporter gene and anxiety disorders. AB - 1. A polymorphism of the variable-number-tandem-repeat (VNTR) in the second intron of the serotonin transporter (ST) gene, which has been reported to be associated with major depression, was studied in anxiety disorders. 2. The VNTR of the human ST gene was compared between 103 patients with anxiety and 106 controls. 3. The frequency of the allele containing 12 copies of the VNTR element (STin2.12) was significantly higher in the combined patient group (p = 0.027), and among patients with OCD (p = 0.0326), and GAD (p = 0.0123), in comparison with in controls. 4. The presence of the STin2.12 allele was significantly associated with the risk of combined anxiety disorders (odds ratio = 2.06, 95% CI 1.09-3.90), OCD (10.2, 1.34-77.4), and GAD (3.61, 1.23-10.6). PMID- 10368857 TI - Antipsychotic drugs affect glucose uptake and the expression of glucose transporters in PC12 cells. AB - 1. Adherence of the PC12 cell line to poly-l-lysine (PLL) on tissue culture dishes stimulated glucose transport into the cells. Fluphenazine, chlorpromazine, clozapine and haloperidol inhibited glucose uptake in this system after a short (30 min) preincubation with drug. The IC50's for this effect were typically in the range of 5-40 microM. 2. Following longer exposures of the drugs (24 hr), there was a significant increase (approximately 3-fold) in the cellular levels of the glucose transporter (GLUT) isoforms, GLUT1 and GLUT3. 3. Long-term incubation (48 hr), especially with the phenothiazine drugs, was accompanied by a marked reduction in cell growth and proliferation. The rank ordering of the potencies of the drugs was essentially the same for these various effects: fluphenazine > chlorpromazine > clozapine approximately haloperidol. 4. It is suggested that the effects on glucose transport reported here may complicate the interpretation of positron emission tomography (PET) studies that rely on the uptake of radiolabeled glucose analogs to measure the physiological response to these drugs. PMID- 10368858 TI - Study of the modulatory activity of BZ (omega) receptor ligands on defensive behaviors in mice: evaluation of the importance of intrinsic efficacy and receptor subtype selectivity. AB - 1. This study examined the hypothesis that the anxiolytic effects of benzodiazepine (BZ (omega)) receptor ligands may be associated with actions at a defined receptor subtype and/or their level of intrinsic activity using the mouse defense test battery. 2. This test has been designed to assess defensive reactions of Swiss mice confronted with a natural threat (a rat) and situations associated with this threat. Primary measures taken before, during and after rat confrontation were escape attempts, flight, risk assessment and defensive threat and attack. 3. The drugs used were the non-selective BZ (omega) receptor full agonist diazepam, the non-selective BZ (omega) receptor partial agonist bretazenil and the beta-carboline abecarnil which acts as a full agonist on GABAA receptors containing the alpha 1- and the alpha 3-subunits and as a partial agonist at receptors containing the alpha 2- and the alpha 5-subunits. The drugs were given alone and diazepam was co-administered with either bretazenil or abecarnil. 4. When administered alone, diazepam attenuated several defensive responses including risk assessment activities, defensive threat/attack reactions upon forced contact with the rat and escape attempts following the removal of the rat from the apparatus. Unlike diazepam, bretazenil was devoid of significant activity on defense and abecarnil displayed depressant activity. 5. Bretazenil blocked all behavioral effects of diazepam on defense behaviors. The co administration of diazepam and abecarnil produced a behavioral profile similar to that observed when diazepam was administered alone, indicating that abecarnil did not influence the effects of diazepam on defense. By contrast, diazepam completely antagonized the sedative effects of abecarnil. 6. These findings indicate that only BZ (omega) ligands with high intrinsic efficacy at all BZ (omega) receptor subtypes display clear and specific effects on defensive behaviors in mice, and suggest that GABAA receptors containing the alpha 3 subunit might represent the primary target involved in the modulatory action of diazepam on defensive behaviors. PMID- 10368859 TI - Behavioral neurochemistry reveals a new functional dichotomy in the shell subregion of the nucleus accumbens. AB - 1. The behavioral and neurochemical effects produced by the direct infusion of amphetamine by reverse microdialysis into either the core or shell of the nucleus accumbens were studied across the anteroposterior axis of this nucleus. 2. Amphetamine (0.05; 0.10; 0.50; 1.00 microM) produced a dose-dependent increase in locomotor activity after microinfusion into either the rostral shell, caudal shell or core of the nucleus accumbens. However, the amphetamine-induced locomotor activating effect, was significantly higher in the rostral shell of the nucleus accumbens compared with both the caudal shell and core. 3. The lowest concentrations of amphetamine produced an equipotent decrease in dialysate dopamine in either the rostral shell, caudal shell, or core. At 1.0 microM, however, amphetamine selectively increased dopamine in the rostral shell. In contrast, the highest dose of amphetamine significantly increased dialysate serotonin levels over baseline only in the caudal shell of the nucleus accumbens. 4. These results demonstrate the preferential effect of amphetamine on dopamine in the rostral shell and serotonin in the caudal shell subterritory of the nucleus accumbens. PMID- 10368860 TI - Potentiation of synaptic transmission by (S)-3,5-dihydroxy phenylglycine in the rat dentate gyrus in vitro: a role for voltage dependent calcium channels and protein kinase C. AB - 1. The authors have previously shown that direct activation of metabotropic glutamate receptors (mGluRs) by (S)-3,5-dihydroxyphenylglycine ((S)-DHPG) can induce a long-lasting potentiation of synaptic transmission in the rat dentate gyrus in vitro. Here the authors provide further characterisation of this agonist induced potentiation. 2. Field excitatory post-synaptic potentials were recorded from the denate gyrus of rat hippocampal slices prepared by standard methods. 3. (S)-DHPG (40 microM) induced a significant potentiation of the field EPSP slope (148.6 +/- 4.3% compared to controls, n = 5), which occluded tetanically-induced LTP. 4. This potentiation was inhibited by the PKC inhibitors staurosporine (0.1 microM) and H-7 (100 microM) and by the voltage dependent Ca2+ channel (VDCC) blockers NiCl2 (50 microM) and nifedipine (20 microM). 5. The mGluR5 specific agonist (RS)-2-Chloro-5-Hydroxyphenylglycine (CHPG) did not induce a potentiation when applied to slices at concentrations from 20 microM to 1 mM indicating that the (S)-DHPG potentiation may be mediated through group I subtype 1 mGluRs. 6. In conclusion the (S)-DHPG-induced potentiation observed in our studies may be PKC dependent and is likely to be mediated through both T/L subtype VDCC and mGluR1 subtype receptors. PMID- 10368861 TI - Circadian rhythm of stereotyped complex behaviours in rats in environmental lead exposure. AB - 1. Stereotyped complex behaviours are present in a number of psychotic illnesses, neurological diseases and even can be generated in response to chemical environment (e.g., drugs or toxins). 2. The circadian rhythm of complex behaviours such as, rearing, preening, scratching and biting/licking was evaluated in an open-field situation in rats exposed to lead (2% lead acetate in drinking water for 30 days). 3. The circadian rhythm of rearing patterns showed depressions from 2 to 14 hr on day 3 and 13, and from 2-6 hr on day 23 (it elevated at 10 hr), whereas increased pattern was apparent at all test periods (except at 6 hr) on day 30. 4. Increased responses of circadian rhythm of preening behaviour were obtained at 18 hr (with decrease at 22 hr) on day 3, at 6 and 14-22 hr on day 13 and, at all the test periods on day 23 (except at 6 hr) and on day 30. 5. The rhythmic patterns of the scratching behaviour showed elevations at each test period as observed on day 3, 13, 23 and 30. The responses in lead-intoxicated rates, however, showed depressions in the light-period and augmentations in the dark-period. 6. The biting/licking behaviours indicated increased patterns of the circadian rhythm attaining a parabolic response, which were inconsistent to the scratching behaviour patterns. Amongst lead-intoxicated rats this behaviour exhibited depressed responses in light-period, whereas in dark-period it showed elevations. PMID- 10368862 TI - Intra-striatal phencyclidine inhibits N-methyl-D-aspartic acid-stimulated increase in glutamate levels of freely moving rats. AB - 1. The authors investigated the effect of local phencyclidine (phenylcyclohexylpiperidine, PCP) on extracellular levels of glutamate and gamma amino butyric acid (GABA) in rat striatum using in vivo microdialysis. 2. Intrastriatal infusion of PCP (1 mM) via a microdialysis probe did not alter the basal extracellular levels of either glutamate or GABA. Addition of N-methyl-D aspartic acid (NMDA; 0.2, 0.5 and 1 mM) to the perfusion medium resulted in a dose-dependent increase in extracellular levels of glutamate. 3. Intrastriatal infusion of tetrodotoxin (0.1, 1, 10 microM), a highly selective blocker of voltage-dependent sodium channels, significantly attenuated the NMDA-stimulated release of glutamate, suggesting that NMDA-evoked release of glutamate originated from the neuronal pool and that the increase of striatal glutamate level was regulated indirectly via NMDA receptors. 4. The NMDA-induced release of glutamate was reduced significantly by pretreatment with local PCP (1 mM). Dizocilpine (MK801; 0.2 mM), a non-competitive NMDA antagonist, completely inhibited the NMDA stimulated release of glutamate. 5. These results suggest that, in the striatum, PCP inhibits corticostriatal glutamatergic neurotransmission by inhibiting the release of glutamate probably via postsynaptic NMDA receptors. PMID- 10368864 TI - Backward masking in bipolar affective disorder. AB - 1. When an informational stimulus, the target, is followed closely in time by a non-informational stimulus, the mask, the visual system's processing of the informational stimulus is disturbed. This disturbance is known as backward visual masking. 2. Transient and sustained visual pathways detect different characteristics of a visual stimulus, at different times in early visual information processing, and have unique anatomic distribution with regard to retinal origin, thalamic and cortical projections. 3. Backward masking occurs by two mechanisms. Interruption occurs when activity in the transient channels of the mask disrupt activity in the sustained channels of the target. Integration occurs when activity in the sustained channels of the mask disrupt activity of the sustained channels of the target. 4. Characteristics of the mask--energy, location, or the time presented after the target--can be altered to enhance interruption or integration. Interruption is a bell-shaped function of, and integration is an exponential function of, visual performance and interstimulus interval. 5. An impairment in backward masking is present in bipolar subjects during manic episodes, is not related to the presence of psychotic symptoms, and persists when mania resolves. Lithium appears to have a detrimental effect on backward masking. PMID- 10368863 TI - A neuroanatomic model for depression. AB - 1. Emotion and mood, once thought to be governed solely by the limbic system of the brain, now are thought to be influenced by numerous nonlimbic central nervous system structures as well. 2. The present review discusses several important brain structures and neuroanatomic pathways thought to be involved in affect and mood disorders, including the amygdala, frontal neocortex, cingulate gyrus, basal ganglia, and the monoamine systems. 3. The authors propose a specific neuroanatomic model for depression that emphasizes that a distributed system of extensively interconnected CNS structures mediates emotion and affect. PMID- 10368865 TI - Tests of executive functioning predict scores on the MacAndrew Alcoholism Scale. AB - 1. Previous work reported that tests of executive functioning (EF) predict the risk of alcoholism in subject populations selected for a "high density" of a family history of alcoholism and/or the presence of sociopathic traits. The current experiment examined the ability of EF tests to predict the risk of alcoholism, as measured by the MacAndrew Alcoholism Scale (MAC), in outpatient subjects referred to a general neuropsychological testing service. 2. Sixty-eight male and female subjects referred for neuropsychological testing were assessed for their past drinking histories and administered the Wisconsin Card Sorting Test, the Wechsler Adult Intelligence Scale-Revised, the Trails (Part B) Test, and the MAC. Principal Components analysis (PCA) reduced the number of EF tests to two measures, including one that loaded on the WCST, and one that loaded on the Similarities, Picture Arrangement, and Trails tests. Multiple hierarchical regression first removed the variance from demographic variables, alcohol consumption, and verbal (i.e., Vocabulary) and non-verbal (i.e., Block Design) IQ, and then entered the executive functioning factors into the prediction of the MAC. 3. Seventy-six percent of the subjects were classified as either light, infrequent, or non-drinkers on the Quantity-Frequency-Variability scale. The factor derived from the WCST on PCA significantly added to the prediction of risk on the MAC (p = .0063), as did scores on Block Design (p = .033). Relatively more impaired scores on the WCST factor and Block Design were predictive of higher scores on the MAC. The other factors were not associated with MAC scores. 4. These results support the hypothesis that decrements in EF are associated with risk factors for alcoholism, even in populations where the density of alcoholic behaviors are not unusually high. When taken in conjunction with other findings, these results implicate EF test scores, and prefrontal brain functioning, in the neurobiology of the risk for alcoholism. PMID- 10368866 TI - Specific binding of 3H-spiperone to peripheral blood cells: relevance for the interpretation of binding studies in psychiatric disorders. AB - 1. There is an ongoing discussion regarding the elevated binding of 3H-spiperone to lymphocytes of schizophrenic patients. Several authors described an atypical binding pattern with a saturable high-affinity and a nonsaturable binding site both displaceable by (+)-butaclamol. Recent findings are still controversial (Fartacek et al., 1997; Wodarz et al., 1996), possibly due to methodological differences. The authors investigated 3H-spiperone binding to different peripheral blood cells including B- and T-lymphoblastoids. 2. B-lymphocytes (KD = 0.081 nM; Bmax = 0.46 x 10(-15) mol/10(6) cells) and macrophages (KD = 0.1-1 nM, Bmax = 2.44 x 10(-15) mol/10(6) cells) are characterized by a minor but saturable binding of 3H-spiperone in a concentration range between 0.5 and 1 nM. Above 1 nM, only non-saturable binding was measurable. Interestingly, Epstein-Barr virus (EBV) transformed lymphoblastoids (KD = 0.13 nM) have nearly the same affinity for 3H-spiperone as native B-cells, but an increased number of binding sites (Bmax = 1.76 x 10(-15) mol/10(6) cells). 3. Membranes from B-lymphoblastoids displayed a saturable binding in a concentration between 0 and 1.8 nM of 3H spiperone (KD = 0.5 nM and Bmax = 1.72 x 10(-15) mol/mg protein). Extraction with 1% digitonin resulted in a similar binding characteristic (KD = 0.17 nM and Bmax = 1.97 x 10(-15) mol/mg protein). 4. T-cells, granulocytes and MOLT-3-cells did not show is a saturable binding even not at high concentrations of 3H-spiperone. 5. The pharmacological profile of the high-affinity 3H-spiperone binding site is clearly different from the dopamine D2 and D4, serotonin 5-HT2 histamine H1 and noradrenergic alpha1 and alpha2 receptor, respectively. 6. In summary, the results suggest that spiperone binding studies to enriched lymphocytes of psychiatric patients should be interpreted cautiously. Variable amounts of leucocytes might result in a higher proportion of nonsaturable, butaclamol displaceable spiperone binding with relevance for the calculation of KD and Bmax of the saturable high-affinity site. Interestingly, homogenous B-lymphoblastoid cell lines have the same binding characteristics as native B-lymphocytes and therefore should be recommended for characterization of 3H-spiperone binding sites. PMID- 10368867 TI - Cognitive impairment and cerebral atrophy in "heavy drinkers". AB - 1. Aim of the work was to verify the following three hypotheses in alcoholics: a) right hemisphere; b) diffuse brain deficit; c) anterior brain deficit, by means of a neuropsychological and a neuroradiological assessment. 2. 15 alcoholic right hand male subjects and 15 matched controls were enrolled in the study. 3. Specifically designed neuropsychological testing was performed to investigate logical abilities, selective attention and memory. 4. Neurological investigation was performed by a standard CT scan to assess the degree and localization of brain damage. 5. Alcoholics performed worse than controls on some neuropsychological tests, i.e. Attention Matrices Test, Verbal Judgement Test, Forward Digit Span, Story Recall and Remote Memory Test. The analysis of variance adjusted by the attentional score showed no significant differences between alcoholics and controls. 6. Neuroradiological data showed a preeminent and a more frequent atrophy of the frontal region. 7. No correlations emerged between neuropsychological and neuroradiological data. 8. In conclusion, the hypothesis of anterior brain deficit seems to be confirmed by our study. PMID- 10368868 TI - Absence of correlation between amobarbital distribution as assessed with SPECT brain perfusion imaging and behavioral manifestations during the intracarotid amobarbital procedure (Wada test). AB - 1. The IAP is used presurgically in patients with temporal lobe epilepsy to predict the effects on LTM and language of the planned temporal lobectomy. This prognosis presumes that a similar pattern of perfusion will result in anesthesia of the same cerebral regions in most patients. 2. Coinjection of Tc-99m HMPAO with the barbiturate during the IAP has been used to ascertain whether this actually is true, with variable results. Moreover, most studies document only unilateral IAPs and do not report on behavioral performance. 3. The authors coinjected Tc-99m HMPAO and amobarbital in 33 IAPs from 18 patients (15 injected bilaterally, 3 unilaterally) to clarify this and to evaluate the relationship of the perfusion pattern to behavioral performance; SPECT results were also compared to angiographic evaluation obtained at the time of catheter placement. 4. SPECT perfusion data was rated for presence/absence and intensity of perfusion to the ACA, MCA, PCA territories and to H, i or c to the injection site. V, STM and LTM were graded according to a standardized protocol. 5. MCAi was perfused in 100% of cases, ACAi in 91%, PCAi in 21% and Hi in only 6%. Cross-over flow was shown in 9 studies; 50% of the patients in whom both sides were injected (on different days) had crossover, involving the ACAc territory in 80% of cases. As expected, injection on the non-ES was associated with a significantly worse LTM performance than on the ES (p = 0.006). There was no relationship between the perfusion pattern and the V level of the patients (a potential confounding variable in memory/language evaluation) during IAP, nor between perfusion pattern and LTM. STM was significantly adversely affected by the presence of crossover perfusion. Angiography in general overestimated the extent of cerebral perfusion demonstrated by SPECT, most probably because of the markedly different injection conditions. 6. Despite the best efforts to standardize injections, the perfusion pattern has been mostly unpredictable in the patients. Moreover, it has little bearing on their behavioral performance, except for the prediction of poor STM performance (the clinical implications of this remaining dubious). Marked LTM alterations after non-ES injections confirm remote hippocampal effects in the presence of only rare direct perfusion of that region. Tc-99m HMPAO/Amobarbital coinjection was unhelpful from a clinical perspective, most probably because a large part of the effects of amobarbital arise from deafferentation of regions not directly perfused by the anesthetic agent. PMID- 10368869 TI - Glucose tolerance and serum insulin levels in an animal model of obesity induced by sub-acute or chronic administration of antipsychotic drugs. AB - 1. To assess the role of insulin in the development of the obesity induced by antipsychotic drugs, a glucose tolerance test was conducted in female rats after 0 or 30 days of sulpiride administration. 2. At day 10, the area under the glucose curve did not differ between sulpiride and vehicle-treated rats, however, the area under the insulin curve was significantly decreased by sulpiride. At day, 30 the insulin response was similar in both groups, but the area under the glucose curve was significantly lower in the sulpiride-treated rats. 3. The results suggest that insulin resistance and hyperinsulinemia are not involved in the development and maintenance of the obesity induced by sulpiride. Contrarily, these findings may be related to an increased insulin sensitivity. 4. The absence of hyperinsulinemia and insulin resistance indicates important differences between primary obesity in humans and rodents and this model of drug-induced excessive weight gain. PMID- 10368870 TI - GABAB receptors are involved in the control of acute opiate withdrawal in isolated tissue. AB - 1. The effects exerted by GABAB receptor agonists and antagonists on the acute opiate withdrawal induced by mu and k receptor agonists were investigated in vitro. 2. Following a 4 min in vitro exposure to morphine (less selective mu agonist), DAGO (highly selective mu agonist) and U50-488H (highly selective k agonist) the guinea-pig isolated ileum exhibited a strong contracture after the addition of naloxone. 3. The selective GABAB receptor agonist, baclofen, at concentration of 5 x 10(-9) - 1 x 10(-8) - 5 x 10(-8) M was able to reduce dose dependently the naloxone-induced contracture after exposure to mu (morphine and DAGO) and k (U50-488H) opiate agonists. 4. Pretreatment with phaclofen (5 x 10( 9) - 1 x 10(-8) - 5 x 10(-8) M), a selective GABAB receptor antagonist, inhibited dose dependently baclofen antagonism on responses to both mu and k agonists. 5. The results of our experiments indicate that GABAB receptors are involved in the control of opiate withdrawal in vitro, confirming an important functional interaction between the GABAergic system and opioid withdrawal. PMID- 10368871 TI - Evidence of sex related differences in the effects of calcium channel blockers on neuroleptic-induced catalepsy in mice. AB - 1. Calcium channel blockers (CCBs) are reported to affect extrapyramidal motor behavior in mammals. Since sex related differences are a common feature in the pharmacological properties of several centrally active drugs, the authors decided to investigate the effects of verapamil (VER), flunarizine (FLU) and nimodipine (NIM), three pharmacologically different CCBs, on neuroleptic-induced catalepsy in male and female albino mice. 2. Catalepsy was induced with haloperidol (0.75 mg/kg, i.p.) and measured at 30-min intervals by means of a bar test. Drugs (or appropriate vehicle, for the controls) were injected i.p. 20 min before haloperidol, with each animal being used only once. 3. VER (1, 5 and 10 mg/kg) did not significantly affect catalepsy in male mice. In females, however, a significant attenuation of catalepsy was found at the two higher doses. 4. FLU (1, 5 and 10 mg/kg) did not significantly affect catalepsy in male mice, whilst a significant attenuation was observed in females with the doses of 1 and 5 mg/kg (but not with the dose of 10 mg/kg). 5. NIM (3, 10 and 30 mg/kg) potentiated neuroleptic-catalepsy in males at the doses of 10 and 30 mg/kg. In females, however, only the higher dose of NIM caused a potentiation of catalepsy. 6. These results demonstrate the existence of sex related differences in the extrapyramidal effects of CCBs in mice. Further, this sex related effect might depend, among other factors, on the particular channel involved. PMID- 10368872 TI - Further investigations of the serotonergic properties of the ibogaine-induced discriminative stimulus. AB - 1. 5-HT3, 5-HT2C, and 5-HT1A receptor ligands were assessed in rats trained to discriminate ibogaine from water. 2. Significant ibogaine-appropriate responding was observed following treatment with the 5-HT2C agonists MK-212 (79.6%) and mCPP (76.4%). This substitution was completely antagonized by metergoline, an agent with 5-HT2C antagonist properties. However, metergoline was ineffective against ibogaine itself. This suggests that although ibogaine may act as an agonist at 5 HT2C receptors, this interaction is not essential to its discriminative cue. 3. Neither the 5-HT3 agonist, mCPBG (44.3%), nor the 5-HT3 antagonist, ondansetron (48.9%) substituted for ibogaine. Likewise, the 5-HT1A agonist 8-OH-DPAT (34.7%) and the 5-HT1A antagonist WAY-100635 (30.1%) failed to substitute. Furthermore, WAY-100635 failed to antagonize the ibogaine cue. 4. Unlike 5-HT2C receptors, 5 HT1A and 5-HT3 receptors do not appear to be involved in the ibogaine stimulus. PMID- 10368873 TI - Behavioral profile of 3,4-methylenedioxy-methamphetamine (MDMA) in agonistic encounters between male mice. AB - 1. The effects of acute administration of 3, 4-methylenedioxymethamphetamine (MDMA), a synthetic amphetamine derivative (0.5-20 mg/kg, i.p.) on agonistic behavior elicited by isolation in male mice were examined. 2. Individually housed mice were exposed to anosmic "standard opponents" 30 min after MDMA injection, and the encounters were videotaped and evaluated using an ethologically based analysis. 3. MDMA (5-20 mg/kg) exhibited a behavioral profile characterized by a reduction of aggression (threat and attack) without a concomitant increase of immobility, accompanied by a decrease of social investigation and a increment of exploration from a distance, avoidance/flee and defense/submission behaviors. 4. This ethopharmacological profile might suggest an anxiogenic-like activity of MDMA in albino male mice. PMID- 10368874 TI - Effects of dothiepin on delayed conduction produced by ventricular arrhythmia in the canine heart after myocardial infarction. AB - 1. In order to clarify the arrhythmogenic effects of antidepressants, the authors examined the effects of dothiepin and amitriptyline on the ventricular activation time (VAT), effective refractory periods(ERP) and incidence of arrhythmias induced by programmed electrical stimulation(PES) in the dog heart in situ after myocardial infarction. 2. Myocardial infarction was produced by two-stage ligation of the left anterior descending coronary artery. Seven days after ligation, bipolar electrodes were sutured on the ventricular surface of the infarcted and normal zones to apply an electrical stimulation or record ventricular activation. An electrical stimulation with coupling interval 250 or 180 ms was applied on the ventricular surface, and AT was measured. 3. Dothiepin at doses of 1-3 mg/kg increased the heart rate. The VAT of coupling interval 180 ms in the infarcted zone was increased by the administration of 3 mg/kg dosulepin. Dothiepin at 3 mg/kg increased the incidence of ventricular arrhythmias induced by PES. 4. Amitriptyline, at doses of 1-3 mg/kg, significantly increased the heart rate. Amitriptyline increased the VAT dose- and frequency-dependently(2,3 mg/kg zone), and prolonged the ERP and QT c interval. Amitriptyline at doses of 1-3 mg/kg increased the incidence of ventricular arrhythmias by PES. 5. These results indicate that dothiepin, 1-3 mg/kg, has lesser effects on cardiac delayed conduction produced by ventricular arrhythmia than amitriptyline. PMID- 10368875 TI - Possible association between delusional disorder, somatic type and reduced regional cerebral blood flow. AB - 1. A 78-year-old female with DDST and pain disorder was treated by clomipramine 20-100 mg/day. The hypochondriacal delusion was completely resolved, while the pains were partially resolved. 2. The SPECT using Xe-133 taken at the early stage of clomipramine treatment, when she still had hypochondriacal delusions, showed markedly reduced rCBF in the temporal and parietal lobes, with predominance on the left hemisphere. Meanwhile, the SPECT taken after resolution of the hypochondriacal delusions showed a marked improvement in the reduced rCBF. 3. This report suggests that DDST has some association with reduced rCBF in the temporal and parietal lobes. PMID- 10368876 TI - Smoking-induced elevations in blood carboxyhaemoglobin levels. Effect on maximal oxygen uptake. AB - Many people engage in physical activity to reduce their cardiovascular risk associated with smoking. These people should be made aware of the metabolic and cardiorespiratory changes induced by chronic and acute smoking and, in particular, the exercise ramifications of increased levels of blood carbon monoxide (CO). Smoking-induced elevations in the CO content of the blood can reduce exercise tolerance and maximal aerobic capacity. Smoking also increases the reliance upon glycolytic metabolism during exercise. Together, these factors contribute to earlier fatigue in smokers compared with nonsmokers who exercise. Similar effects upon exercise tolerance are noted in those who inhale environmental tobacco smoke. PMID- 10368877 TI - Exercise stress testing. An overview of current guidelines. AB - Exercise stress testing (ET) is an inexpensive noninvasive tool that provides valuable cardiopulmonary information in healthy and diseased populations. It is most commonly used for diagnosing coronary artery disease (CAD) and developing appropriate exercise prescriptions (EP). With its widespread use and application, it is imperative that safe and appropriate guidelines and procedures are used, as there are a number of risks associated with testing in a population with or suspected of having CAD. The focus should be on the patient's safety: personnel must be properly trained and aware of all emergency procedures, contra indications for ET and indications for test termination must be strictly adhered to. Three main types of testing are prevalent: submaximal, maximal and maximal utilising gas exchange. The maximal test is most commonly used, and the submaximal is appropriate for hospitalised patients. Gas exchange data is essential when assessing congestive heart failure and timing for heart transplantation. ET is commonly performed using a treadmill or a bicycle ergometer. The treadmill provides a more familiar exercise modality and has been shown to have greater diagnostic sensitivity than the bicycle ergometer; it is, however, more expensive and requires more space in the testing room. The bicycle ergometer is more appropriate for those individuals who are severely obese or have problems with extended periods of walking. Regardless of the modality used, an appropriate exercise protocol should be used. In North America, the Bruce protocol is the most common. However, the Bruce protocol, and others that estimate exercise capacity based on equations, tend to overestimate exercise capacity. They may be too demanding for those with limited exercise capacity, and too long for those with high exercise capacity. For these people, an exercise protocol that reaches maximal capacity in 8 to 12 minutes using smaller increments in workload should be considered. Once completed, the results of ET needs to be correctly interpreted. This includes reviewing the test results while considering the patient's history, medications and indication for the test. ET can also be used to develop an EP for the participant. An EP should take into account the intensity, modality of exercise, frequency and duration, as well as being realistic for the individual and the goals to be achieved. All the information from the test results and the pre-test examination should be presented in a report that also includes the advised EP. PMID- 10368878 TI - Oxygen uptake kinetics during exercise. AB - The characteristics of oxygen uptake (VO2) kinetics differ with exercise intensity. When exercise is performed at a given work rate which is below lactate threshold (LT), VO2 increases exponentially to a steady-state level. Neither the slope of the increase in VO2 with respect to work rate nor the time constant of VO2 responses has been found to be a function of work rate within this domain, indicating a linear dynamic relationship between the VO2 and the work rate. However, some factors, such as physical training, age and pathological conditions can alter the VO2 kinetic responses at the onset of exercise. Regarding the control mechanism for exercise VO2 kinetics, 2 opposing hypotheses have been proposed. One of them suggests that the rate of the increase in VO2 at the onset of exercise is limited by the capacity of oxygen delivery to active muscle. The other suggests that the ability of the oxygen utilisation in exercising muscle acts as the rate-limiting step. This issue is still being debated. When exercise is performed at a work rate above LT, the VO2 kinetics become more complex. An additional component is developed after a few minutes of exercise. The slow component either delays the attainment of the steady-state VO2 or drives the VO2 to the maximum level, depending on exercise intensity. The magnitude of this slow component also depends on the duration of the exercise. The possible causes for the slow component of VO2 during heavy exercise include: (i) increases in blood lactate levels; (ii) increases in plasma epinephrine (adrenaline) levels; (iii) increased ventilatory work; (iv) elevation of body temperature; and (v) recruitment of type IIb fibres. Since 86% of the VO2 slow component is attributed to the exercising limbs, the major contributor is likely within the exercising muscle itself. During high intensity exercise an increase in the recruitment of low-efficiency type IIb fibres (the fibres involved in the slow component) can cause an increase in the oxygen cost of exercise. A change in the pattern of motor unit recruitment, and thus less activation of type IIb fibres, may also account for a large part of the reduction in the slow component of VO2 observed after physical training. PMID- 10368879 TI - Effect of fluvastatin in combination with moderate endurance training on parameters of lipid metabolism. AB - OBJECTIVE: To establish whether patients receiving the cholesterol synthesis enzyme inhibitor fluvastatin 20 mg/day could obtain an additional improvement in their lipid pattern as a result of physical endurance training. DESIGN: This was an observational study using a before- and after-treatment comparison of fitness and lipid parameters in outpatients with dyslipidaemia who undertook an exercise programme with or without treatment with a lipid-lowering drug. STUDY PARTICIPANTS: Participants were 18 sedentary [maximum oxygen uptake (VO2max) < 30 ml/kg bodyweight per minute] men (age range 38 to 65 years) with dyslipidaemia but without overt cardiovascular disease. INTERVENTIONS: All participants undertook a 1-hour bout of endurance training twice a week for 3 months. The training involved a circuit using various ergometers, with continuous monitoring of pulse rate, at an exercise intensity of 2 to 3 mmol/L lactate. The control group (n = 6) received no drug treatment; they completed the training programme only. The pretreatment group (n = 6) comprised participants who had already been treated with fluvastatin 20 mg/day for at least 3 months before beginning the training programme. The treatment group (n = 6) received fluvastatin 20 mg/day from the beginning of the training programme. All participants were required to comply with the exercise programme and with a standardised carbohydrate-loaded diet together with restriction of alcohol consumption to a maximum of 20 ml/day. RESULTS: In the control group, increased physical activity alone reduced serum triglyceride (TG) levels (-24.7%) and increased serum high density lipoprotein cholesterol (HDL-C) levels (+19.3%). There was a smaller effect on serum low density lipoprotein-cholesterol (LDL-C) levels (-12.8%). Similar but smaller effects were observed in the pretreatment group (i.e. patients previously treated with fluvastatin): TG -12.88%, HDL-C +13.81%, LDL-C -8.7%. Marked changes were observed in the treatment group: TG -33.1%, HDL-C +34.7%, LDL-C -40.5%, total cholesterol -30.5%. CONCLUSIONS: A reduction of serum LDL-C level in the target range of -30 to -40% cannot be achieved by this intensity of training alone. In combination with fluvastatin 20 mg/day, however, the positive effects on lipid metabolism are potentiated. Thus, treatment with fluvastatin combined with moderate endurance training is a rational mode of therapy, particularly in patients with a highly pathological lipid profile. PMID- 10368880 TI - Recommendations for the diagnosis of traumatic meniscal injuries in athletes. AB - It has always been difficult to develop a method of correctly evaluating knee injuries and, in turn, to devise the appropriate rehabilitation programme. Flawless diagnosis of meniscal injury is necessary, considering the diverse consequences of this injury for patients, and even more so in relation to athletes, bearing in mind the intensified physical demands on their bodies. There is no doubt that an accurate and concise clinical evaluation of patients with injuries to the knee is the basis for an exact diagnosis and successful treatment. The use of noninvasive methods, such as magnetic resonance imaging, in addition to clinical evaluation is recommended because of their high accuracy and negative predictive value. The use of invasive methods, such as arthroscopic operations, should be restricted to treatment, and not be used for diagnosis. PMID- 10368881 TI - [Nikolai Sergeevich Korotkov (1874-1920)]. PMID- 10368882 TI - [The possibilities for 2-stage instrumental screening in the detection of thyroid diseases in an endemic focus]. AB - The approbation and comparative assessment of the methods of early active detection of thyroid diseases were performed among the population of the endemic region. The ultrasound examination of the thyroid gland is thought to be the most effective method at the first stage of the screening examination since it is very exact (98%), specific (99%) and sensitive (99%) when detecting both diffuse and nodular alterations of the organ. The possibility to combine the ultrasound examination with the thin-needle aspiration puncture biopsy allows not only to detect the patients and form the groups of risk of probable diseases, but also to make the diagnosis and determine the methods of treatment during the mass examination. PMID- 10368883 TI - [Endoscopic and histological changes in the mucosa of the gastric stump in Helicobacter pylori infection]. AB - Under investigation there were 50 patients with resected stomach at different terms after operations: 37 patients after subtotal resection of the stomach for cancer and 13 patients after resection of 2/3 of the stomach for ulcer disease. The overwhelming majority of the patients with the resected stomach had chronic gastritis of the stump diagnosed endoscopically and histologically. No reliable difference of the endoscopic and histological indices of the state of the gastric stump mucosa was found in relation to the disease which was the cause of resection and to the time since the operation. The presence of Helicobacter pylori in the gastric stump mucosa was found in 37 of 50 examined patients (74%). In the group of patients with Helicobacter pylori in the gastric stump mucosa there were no reliable differences in endoscopic findings but changes in the histological picture were found significantly more often independent of the disease which caused the resection. Atrophic alterations in the gastric stump mucosa after resection in patients with Helicobacter pylori develop in patients of younger age and in earlier terms after operation. PMID- 10368884 TI - [Dearterialization of the liver as a treatment method in hemangiomas]. AB - The article is devoted to the experiences with treatment of hemangiomatous lesions of the liver. Results of palliative treatment of 65 patients by methods of dearterialization of the liver (ligation and embolization of the hepatic artery) are presented. The authors make a conclusion that dearterialization of the liver is a sufficiently effective method of treatment resulting in sclerosing the hemangiomas and substitution of them with the fibrous tissue. It is not followed by the development of pronounced complications and lethality. With clinical manifestations retained after embolization of the hepatic artery it is possible to perform resection of the liver with hemangioma, the operative intervention in this case being accompanied by less blood loss, and it is easier technically than when it is fulfilled without preliminary dearterialization of the liver. PMID- 10368885 TI - [The comparative hemodynamic characteristics in the lower extremities during laparoscopic and traditional interventions on the abdominal cavity organs]. AB - The plethysmographic methods were used for studying the influence of pneumoperitoneum in hemodynamics in lower extremities. It was proved to cause venous stasis in them and constriction of precapillaries in the venous area predominantly. The degree of such influence depends on the strain of the pneumoperitoneum and its duration. This factor should be taken into consideration for prophylaxis of postoperative complications. PMID- 10368886 TI - [The pathogenetic validation of the volume and the technology for the surgical correction of disorders in the musculo-venous pump of the lower extremities in patients with decompensated forms of varicose veins]. AB - The state of the venous, capillary blood circulation and lymph flow was studied in 70 patients with varicose disease at the stage of trophic disorders with the help of photoplethysmography, rheovasography, dopplerography, distal ascending and retrograde femoral, popliteal, talocrural phlebography, lymphography. It was established that a sharp disturbance of the musculo-venous pump function caused decompensation of not only the venous blood circulation but also the lymph flow, while the removal of the subcutaneous veins of the crus is not possible without deterioration of the lymphatic drainage. The authors propose not to ablate the main trunks of the subcutaneous veins on the crus but to perform occlusion of the autovein ablated from the femur. A technology of resection of the pathologically altered vein of the soleus and gastrocnemius muscles and retrograde occlusion of the posterior tibial veins is developed for improving the work of the musculo venous pump of the leg. Good results of treatment were obtained in 84.3% of the patients. PMID- 10368887 TI - [Trauma to the forearm with damage to the major arteries]. AB - The authors made an analysis of surgical methods of treatment of 63 patients and long-term results in the period from 1 to 6 years in 23 of them. The restoration of all the injured anatomical structures of the forearm and, in particular, the restoration (suture, autovenous plasty) of the major arteries and primary suture of the injured nerves are thought to be necessary for good results. PMID- 10368888 TI - [A system for assessing the stability of the osteosynthesis in femoral neck fractures]. AB - Results of experimental investigation of different biomechanical parameters of the system "bone-fixative" and bony fixatives are analyzed in relation to different types of fractures of the femur neck. The data obtained are compared with clinical results of osteosynthesis. On this basis an index system is recommended for choosing the method of treatment of fractures and the particular type of the fixative. PMID- 10368889 TI - [Transdrainage closed ultrasonic cleansing of the abdominal cavity in the prevention and treatment of infectious complications in abdominal gunshot wounds]. AB - Under analysis were results of treatment of 112 wounded with gunshot injuries of the abdomen. The flowing-irrigating aspiration dialysis followed by the closed transdrainage ultrasonic sanitation in combination with a medicinal composition (dioxidine, gentamycine, tripsin) and simultaneous intramuscular injection of solcoseryl and thymalin were used for prophylactics and treatment of infectious complications in the abdominal cavity and in the wound of the abdominal wall. The method of treatment was found to result in less amount of pyo-inflammatory complications (from 34.8 to 2.6%), shorter terms of cleansing the wound from pyo necrotic formations and liquidation of the perifocal inflammatory reaction, shorter duration of the stationary treatment. PMID- 10368890 TI - [Patterns in the development of suppurative complications in acute destructive pancreatitis and the means for their prevention]. AB - Under analysis were the results of conservative and operative treatment of 391 patients. It was shown that purulent complications were more frequent in postoperative and ischemic pancreatitis. Intensive treatment of the enzymatic phase of the disease and reactive phase of severe forms can substantially decrease the frequency of purulent complications and lethality. PMID- 10368891 TI - [The incompetence of the appendiceal stump after appendectomy]. AB - Based on an analysis of results of treatment of 2828 patients with acute appendicitis the authors discuss the causes and give recommendations for prophylactics of incompetence of the vermiform process stump. PMID- 10368892 TI - [The causes for the worsening of treatment results in patients with infected pseudarthroses of the tibia in recent years]. AB - Under analysis were results of treatment of 141 patients with infected pseudoarthroses of the tibia. Among them there were 10 women and 131 men aged from 18 to 62 years. The long-term results were followed-up during the period from 2 to 20 years. The analysis allowed a supposition that a combination of tibia segment resection and plasty with local tissues completed by suturing the wound, drainage and external fixation with the Ilizarov device leads to deterioration of the outcomes. A careful approach to treatment of such patients including the division of treatment into the stages of debridement and reconstruction seems to be more productive. PMID- 10368893 TI - [Simultaneous operations in patients with thyroid diseases]. AB - Twenty-four cases of simultaneous surgical treatment for coexisting diseases of the thyroid gland and other organs are discussed. Such operations were mainly performed for associated nodular goiter (including thyroid cancer) and benign or malignant tumours of other localisations. As a rule surgical interventions were planned beforehand. Only in 2 cases there were urgent operations for coexisting diffuse toxic goiter and complicated ulcer disease. In 4 cases thyroid resection and laparoscopic cholecystectomy were made simultaneously. Immediate and long term outcomes were good. The indications and conditions of performing the operations are discussed. PMID- 10368894 TI - [Intestinal obstruction in disseminated malignant tumors of the female genitalia: the surgical procedure and combined operations]. AB - Results of surgical treatment of 103 patients are presented who had spread malignant tumors of the female genitalia with the involvement of different segments of the intestine and urinary tract, and damaged intestinal passage. Combined operations were performed on 57 patients (55.3%), palliative--46 patients (44.7%) 54 patients were operated on urgently. For the last 10 years resectability has been increased 8.5 times owing to active surgical policy. Immediate lethality after combined operations was 10.5%. Cumulative indices of 3- and 5-year survival were 66% and 53.3% respectively. Lethality and 5-year survival after palliative operations were 30.4% and 13.8% respectively. Combined operations for cancer of female genitalia (both primary and recurrent ones and metastases) are quite reasonable in order to provide the optimum volume of cytoreduction and perform chemo- and radiation treatment. PMID- 10368895 TI - [The efficacy of Neovir in the combined treatment of patients with operable tumors of the gastrointestinal tract]. AB - Under observation there were 32 patients with operable tumors of the gastro intestinal tract. The average age of the patients was (64.8 +/- 7.5) years. Patients of the control group (18 patients were given the routine complex treatment. The main group (14 patients) were additionally given the interferonogenic immunomodulator "Neovir" which resulted in lower risk of the appearance of pyo-septic complications. It was confirmed by positive dynamics of the indices of cellular immunity, intensified production of proinflammatory cytokines and interferon-alpha. PMID- 10368896 TI - [The treatment of patients with acute appendicitis by a short-term stay in the hospital]. AB - An analysis of results of treatment of 102 patients with acute appendicitis has shown that use of non-direct endolymphatic therapy decreases the amount of antibiotics and analgetics administered after surgery, leads to less frequency of postoperative complications and the patients can be discharged from the hospital within the first 6 days after operation. PMID- 10368897 TI - [Intra- and extra-abdominal desmoids in men]. PMID- 10368898 TI - [A primary restorative operation with a favorable outcome in cancer of the transverse colon complicated by penetration into the body of the stomach with the formation of an internal fistula]. PMID- 10368899 TI - [The successful treatment of an anaerobic gas infection of the lower extremity at a district hospital]. PMID- 10368900 TI - [The saving of blood in surgery]. PMID- 10368901 TI - [Low-volume infusion therapy with hyperosmolar sodium chloride solutions in the treatment of acute massive blood loss]. PMID- 10368902 TI - [Single-row intestinal sutures and and the current suture materials in colorectal surgery]. PMID- 10368903 TI - [Ignaz Semmelweis--forefather of antisepsis (on the 180th anniversary of his birth)]. PMID- 10368904 TI - [The great surgeons of Russia at the sources of the conception and development of first aid in Saint Petersburg]. PMID- 10368907 TI - Hormonal regulation of actin and tubulin in an epithelial cell line from Chironomus tentans. AB - The morphogenetic changes in an epithelial cell line from Chironomus tentans that are evoked by molting hormones and molting hormone agonists are accompanied by transient changes in the concentration of actin and beta-tubulin protein and mRNA. As compared to controls, actin protein and mRNA concentrations increase by about 50%, whereas tubulin reaches maxima of 100% increase. The proportion between globular and filamentous actin remains constant after hormone treatment. PMID- 10368908 TI - Individual and developmental differences in disengagement of fixation in early infancy. AB - The current study investigated whether individual and developmental differences in look duration are correlated with the latency for infants to disengage fixation from a visual stimulus. Ninety-four infants (52 3-month-olds, 42 4-month olds) were tested in a procedure that measured ocular reaction time to shift fixation from a central target to a peripheral target under conditions in which the central-target either remained present ("competition" condition) or was removed from the display ("noncompetition" condition). Look duration was correlated with disengagement latency; longer-looking infants were slower than shorter-looking infants to shift fixation to the peripheral target on competition trials, but not noncompetition trials. Results were similar for 3- and 4-month olds, although 3-month-olds showed slower latencies on all trials. Furthermore, long-looking infants were not consistently slower, but rather showed greater variability in their response latencies under conditions that required disengagement of fixation. The results support the position that developmental and individual differences in look duration are linked to the development of the neural attentional systems that control the ability to disengage, or inhibit, visual fixation. PMID- 10368909 TI - The relation of affect to attention and learning in infancy. AB - The relation of positive affect to attention and learning was examined in 5-, 7-, and 9-month-olds (N = 84). Affect and attention were assessed while the infants inspected a photograph. Affect was rated globally, for overall mood, and specifically, for amount of time smiling. Attention was indexed by the duration of the infant's longest (or peak) look, a measure previously linked to differential cognitive performance. At all ages, positive affect (shown by approximately half the infants) was associated with long look durations and slower learning, as assessed on a task in which infants learned to distinguish a familiar face from a series of novel faces. By contrast, neutral affect was associated with short looks and faster learning. Affect and look duration had synergistic effects, in that learning was faster than expected for infants who displayed both short looks and neutral affect. These findings are compatible with adult research that links positive affect to less analytical processing, and provide the first evidence that affect may be associated with the speed of processing differences implicated in short and long looking. PMID- 10368910 TI - Child-directed speech produced by mothers with symptoms of depression fails to promote associative learning in 4-month-old infants. AB - Child-directed (CD) speech segments produced by 20 mothers who varied in self reported symptoms of depression, recorded during a structured play interaction with their 2- to 6-month-old infants, were used as conditioned stimuli with face reinforcers in a conditioned attention paradigm. After pairings of speech segments and faces, speech segments were assessed for their ability to increase time spent looking at a novel checker-board pattern (summation test) using 225 4 month-old infants of nondepressed mothers. Significant positive summation, an index of associative learning, was obtained in groups of infants tested with speech produced by mothers with comparatively fewer self-reported symptoms of depression (Beck Depression Inventory or BDI < or = 15). However, significant positive summation was not achieved using speech samples produced by mothers with comparatively more symptoms of depression (BDI > 15). These results indicate that the CD speech produced by mothers with symptoms of depression does not promote associative learning in infants. PMID- 10368911 TI - Where theories of mind meet magic: the development of children's beliefs about wishing. AB - In two studies, we probed children's beliefs about wishing. In Study 1, we gathered initial data on 50 3- to 6-year-old children's concepts of wishing and beliefs about its efficacy, with both a semistructured interview and a variety of tasks. Results revealed considerable knowledge about wishing in young children, along with an age-related decrease in beliefs about its efficacy. Parents were not found to encourage differently the beliefs of children at different ages, nor were they found to begin actively discouraging such beliefs at any particular age. A moderate relation was found between environmental supports for wishing and children's beliefs in its efficacy. In Study 2, we continued to probe these issues and also address the nature of the broader conceptual context in which children situate their beliefs about wishing. Participants were 92 3- to 6-year old children. Results of this study suggest that children may reconcile beliefs in the efficacy of wishing with knowledge about everyday mental-physical relations by situating these beliefs more within their emerging beliefs about magic than within their theories of mind. PMID- 10368912 TI - The use of trait labels in making psychological inferences. AB - Three studies investigated children's capacity to use trait labels as tools for making inferences about mental states. For example, knowledge that a story character is "nice" as opposed to "mean" could lead to predictions that the character would respond with greater negative affect upon discovering that his or her action had made someone upset. Study 1 (N = 48) examined whether participants (kindergartners, second graders, fifth graders, and adults) would make different psychological inferences based on whether a character was labeled as "nice" versus "mean." Study 2 (N = 30) examined the same issue with 4-year-olds using a simpler methodology. Study 3 (N = 30) extended the results of Study 2, by examining whether describing characters as "shy" versus "not shy" would lead 4 year-olds to make different mental state inferences. Taken together, these findings suggest that even for young children, trait labels can serve as a basis for making nonobvious inferences. Developmental differences are discussed. PMID- 10368913 TI - Caregiving and developmental factors differentiating young at-risk urban children showing resilient versus stress-affected outcomes: a replication and extension. AB - This study tested hypotheses from an organizational-developmental model for childhood resilience. In this model resilience reflects a child's mastery of age salient objectives, in the face of substantial adversity, by drawing on internal and external resources that enhance processes of adaptation specific to each developmental stage. Interviews were conducted with parents of 122 7- to 9-year old urban children exposed to multiple risk factors, 69 classified as resilient and 53 as maladjusted. Consistent with predictions generated by the model: (1) characteristics of a child's caregiving system and early development differentiated children with resilient and stress-affected adaptations; and (2) variables reflecting emotionally responsive, competent parenting were direct, proximal predictors of resilient status and mediators of other caregiver resources such as education, mental health, and relational history. Identified predictors of resilient status, including competent parenting and caregiver psychosocial resources, largely replicated findings from a prior study with sociodemographically comparable 9- to 12-year-old children. PMID- 10368914 TI - Age and gender as determinants of stress exposure, generation, and reactions in youngsters: a transactional perspective. AB - The present study used a contextual and transactional approach to examine age and gender differences in the experience and consequences of life stress in clinic referred preadolescents and adolescents. Eighty-eight youngsters and their parents completed the Child Episodic Life Stress Interview, a detailed semistructured interview assessing the occurrence of stressful events in multiple life domains. Interviews were coded using a contextual threat rating method to determine event stressfulness and dependence. Youngsters also completed the Children's Depression Inventory and the Revised Child Manifest Anxiety Scale to assess self-reported symptoms of depression and anxiety. Consistent with predictions, age- and gender-related patterns of life stress varied across the type and context of stressors. Most notably, adolescent girls experienced the highest levels of interpersonal stress, especially stress and conflict that they generated within parent-child and peer relationships. Preadolescent girls experienced the highest levels of independent stress and conflict in the family context. Adolescent boys experienced the highest levels of noninterpersonal stress associated with self-generated events. Girls demonstrated particular vulnerability to depressive responses to dependent stress. The results build on and extend previous theory and research on age and gender differences in close relationships and stress, and illustrate the value of more refined conceptual models and more sophisticated methodologies in child life stress research. PMID- 10368915 TI - Development of adjustment problems in girls: what syndromes emerge? AB - The development of a broad spectrum of adjustment problems in girls was studied longitudinally from late childhood to early adulthood. A specific interest concerned how well the externalizing-internalizing distinction could explain the data. The sample consisted of about 500 Swedish girls, reasonably representative of the general population. Variable-oriented methods were complemented with person-oriented methods to study syndrome formation at the level of the individual. The results suggested a rather diversified pattern of multi-problem syndromes in late childhood, whereas the syndrome structure in early adolescence was organized around a differentiation between girls with externalizing adjustment problems and girls with peer problems. An externalizing syndrome was found to be stable between late childhood and early adolescence, increasing the risk of severe maladjustment in adulthood. Internalizing problems showed no clear cut continuity with adult maladjustment. Results are discussed in relation to the externalizing-internalizing distinction, which to some extent is called in question. PMID- 10368916 TI - Longitudinal and concurrent relations among temperament, ability estimation, and injury proneness. AB - This study examined longitudinal and concurrent relations between temperament, ability estimation, and injury proneness. Longitudinal assessments of Inhibitory Control were collected through a behavioral battery at toddler (33 months) and preschool ages (46 months). Parent-reported measures of Inhibitory Control and Extraversion also were obtained at those ages. At school age (76 months), children participated in a set of tasks to assess overestimation and underestimation of physical abilities. Parents provided reports of children's temperament and injury history at school age. Results showed that children who were high on Extraversion and low on Inhibitory Control as toddlers and preschoolers tended to overestimate their physical abilities and to have more unintentional injuries at age 6. Children low on Extraversion and high on Inhibitory Control tended to underestimate their physical abilities. Implications for injury prevention are discussed. PMID- 10368917 TI - The role of an early intervention on enhancing the quality of mother-infant interaction. AB - The study examines an intervention designed to influence mothers' sensitive responsiveness toward their infant by presenting information about the newborn's competence to interact and promoting affectionate handling and interaction with the infant. Thirty-six primiparous mothers and their newborn infants participated in the study. On day 2/3 after delivery, mother-infant dyads were assigned to either: (1) an experimental group that received an intervention program designed to enhance mother-infant interaction; or (2) a control group that was presented with an intervention that emphasized basic caregiving skills. One month later an observation was undertaken in the home to assess mother-infant synchronous and asynchronous co-occurrences during free-play and infant bathing. The enhancement group showed a reliably greater frequency of co-occurrences involving vocal exchanges, looking to the partner, and physical contact. There also were differences in mothers' responsiveness to infant crying and involuntary responses. The findings show that even a modest videotaped early intervention can enhance mothers' sensitive responsiveness to the infant. PMID- 10368918 TI - Predicting mothers' beliefs about preschool-aged children's social behavior: evidence for maternal attitudes moderating child effects. AB - Maternal beliefs about children's social behavior may be important contributors to socialization and development, but little is known about how such beliefs form. Transactional models suggest that children's characteristics may influence parents. At 2 years of age, the shy and aggressive behaviors of 65 toddlers (28 females) were observed during interactions with an unfamiliar peer; as well, mothers described the extent to which they advocated protective and authoritarian childrearing attitudes. These variables were used to predict mothers emotions, attributions, parenting goals, and socialization strategies in response to vignettes depicting aggressive and withdrawn child behaviors 2 years later. Most child effects were moderated by maternal attitudes or gender effects. Authoritarian mothers of aggressive toddlers were most likely to report high control and anger, to blame their children for aggression, and to focus on obtaining compliance rather than teaching skills to their children. Protective mothers reported that they would use warmth and involvement to comfort withdrawn children, especially their daughters. PMID- 10368919 TI - A prospective study of the effects of marital status and family relations on young children's adjustment among African American and European American families. AB - The present study investigated the effects of divorce and family relations on young children's development prospectively, using an ethnically diverse sample of approximately 300 low-income families. We also were able to examine the moderating effects of ethnicity on child adjustment in always two-parent, to-be divorced, already-divorced, and always single-parent families. Results indicated that to-be-divorced European American and African American families demonstrated higher rates of preschool-age behavior problems, and already-divorced families showed similar trends. Parental conflict and behavior problems accounted for predivorce differences in child behavior problems, whereas rejecting parenting accounted for differences in problem behavior between always single-parent and always two-parent families. The results are discussed in terms of the importance of ethnicity in influencing young, low-income children's adjustment to different family structures. PMID- 10368920 TI - Experiences in after-school programs and children's adjustment in first-grade classrooms. AB - The experiences of 150 children in after-school programs were examined in relation to performance in first grade. Three aspects of program experiences (emotional climate, quality of peer interactions, and program curriculum) were associated with the children's concurrent adjustment at school, controlling for family selection factors. Staff positivity in the after-school programs was associated with boys displaying fewer internalizing and externalizing problems, whereas staff negativity was related to boys obtaining poorer grades in reading and math. Program flexibility was associated with boys having better social skills. More frequent negative interactions with peers in the programs were related to more internalizing and externalizing problems, and poorer social skills at school. Boys who attended programs offering a larger number of different activities had more internalizing and externalizing problems, and poorer grades in reading and math. After-school experiences also were related to girls' behaviors, but associations were less apparent for girls than boys. PMID- 10368921 TI - The impact of after-school peer contact on early adolescent externalizing problems is moderated by parental monitoring, perceived neighborhood safety, and prior adjustment. AB - Unsupervised peer contact in the after-school hours was examined as a risk factor in the development of externalizing problems in a longitudinal sample of early adolescents. Parental monitoring, neighborhood safety, and adolescents' preexisting behavioral problems were considered as possible moderators of the risk relation. Interviews with mothers provided information on monitoring, neighborhood safety, and demographics. Early adolescent (ages 12-13 years) after school time use was assessed via a telephone interview in grade 6 (N = 438); amount of time spent with peers when no adult was present was tabulated. Teacher ratings of externalizing behavior problems were collected in grades 6 and 7. Unsupervised peer contact, lack of neighborhood safety, and low monitoring incrementally predicted grade 7 externalizing problems, after controlling for family background factors and grade 6 problems. The greatest risk was for those unsupervised adolescents living in low-monitoring homes and comparatively unsafe neighborhoods. The significant relation between unsupervised peer contact and problem behavior in grade 7 held only for those adolescents who already were high in problem behavior in grade 6. These findings point to the need to consider individual, family, and neighborhood factors in evaluating risks associated with young adolescents' after-school care experiences. PMID- 10368922 TI - Welfare dynamics, support services, mothers' earnings, and child cognitive development: implications for contemporary welfare reform. AB - This prospective longitudinal study, using data from the National Longitudinal Survey of Youth (NLSY; N = 614), addresses the gap in the research literature regarding the effects of welfare reform on children. Key questions addressed include whether welfare dynamics and support services relevant to welfare reform, both measured across the first 5 years of life, are associated with mothers' earnings in the 6th year and three child cognitive outcomes in the 7th and 8th years: Peabody Individual Achievement Test (PIAT) math and reading scores, and the Peabody Picture Vocabulary Test (PPVT). Welfare dynamics are represented by total time on welfare, degree of cycling on and off welfare, and degree to which welfare and work are combined. Support services measured include three forms of child care (relative, babysitter, and center-based), as well as three forms of human capital supports (child support, job training, and education). Controlling for a range of background factors and for different patterns of welfare use across the first 5 years, small positive associations with mother's earnings were found for child support, education, and job training. Small positive associations also were found between child support and both math and reading scores. Finally positive associations of medium effect size were found between center care and both mothers' earnings and child PPVT scores. Although effect sizes are generally small, the results suggest the potential value of welfare reform approaches that emphasize long-term human capital development. Interactions between welfare dynamics and support services suggest subgroup differences. Specifically, positive effects of support services on earnings are strongest among mothers with higher levels of human capital (higher levels of work while on welfare, lower total time on welfare). Babysitter care appears to have negative effects on both reading and math scores of children whose mothers report low levels of work while on welfare. Implications for welfare reform policy are discussed. PMID- 10368923 TI - Binding of chlorpheniramine enantiomers to human plasma proteins. AB - The in vitro binding of RS-chlorpheniramine to human proteins was studied by equilibrium dialysis. The binding to total plasma proteins and to individual albumin and alpha-glycoprotein acid is stereoselective. (+)S-chlorpheniramine is more extensively bound than its antipode to total plasma proteins (38% vs. 23%), to albumin (20% vs. 15%) and to alpha-glycoprotein acid (23% vs. 5%). PMID- 10368924 TI - Sex differences in stereospecificity of oracin reductases in rat in vitro and in vivo. AB - In vitro and in vivo experiments to investigate possible stereospecific aspects of oracin reduction in relation to rat gender have been conducted. Incubation of oracin with rat microsomes, cytosol, and hepatocytes in the presence of various coenzymes and under aerobic or anaerobic conditions provided evidence for sex differences in the formation of 11-dihydrooracin (DHO) enantiomers. The greatest sex differences were seen in hepatocytes where females showed higher stereospecificity of the reductases than males. While female biotransformation enzymes preferentially generated approximately 82% of (+)-DHO, male enzymes gave only rise to 63% of (+)-DHO. Males displayed higher stereospecificity than females in the microsomal fraction. However, in the cytosolic fraction females exhibited higher stereospecificity than males. Similarly, in in vivo studies, the ratio of (+)- and (-)-DHO in faeces and urine gave no indication of the significant differences between the male and female rat. Enzyme stereospecificity has been defined as preferential formation of the (+)- or (-)-stereoisomer of 11 DHO by the respective enzyme. HPLC quantitative determinations of both enantiomers were performed using a Chiralcel OD-R column as the chiral stationary phase with excellent resolution and stability. PMID- 10368925 TI - A comparison between stereospecificity of oracin reduction and stereoselectivity of oxidation of 11-dihydrooracin enantiomers in vitro in rat and guinea pig. AB - 11-dihydrooracin (DHO) arises from the potential cytostatic drug oracin through the metabolic conversion of its prochiral centre (C11). The participation of reduction enzymes on production of DHO enantiomer under various incubation conditions were tested in rat and guinea pig microsomal and cytosolic fractions. Interesting differences in stereospecificity of oracin reduction enzymes were found. Reduction stereospecificity was further studied on rat and guinea pig isolated hepatocytes. The enantiomers were detected in rat and guinea pig hepatocytes in the (+)/(-) ratio 63/37 and 32/68 respectively. As the differences in the amounts of DHO enantiomers can be caused not only by stereospecificity of oracin reduction but also by subsequent conversion of the enantiomer, stereoselectivity of DHO oxidation to oracin was investigated. Synthetically prepared pure (+)- and (-)-DHO were incubated with rat or guinea pig microsomes and cytosol and with various coenzymes under aerobic or anaerobic conditions. Significant oxidation of DHO to oracin was observed in rat microsomes. This oxidation depends on NADPH and O2 and is stereoselective for (+)-DHO. The formation of oracin in the guinea pig was greater in cytosol than microsomes, but no significant preference for a particular DHO enantiomer was found. PMID- 10368926 TI - Chiral discrimination in the transport of ketoprofen and ibuprofen esters through an aqueous phase mediated by various serum albumins. AB - Serum albumins that act as carriers discriminated between enantiomers of alkyl esters of ketoprofen and ibuprofen in transport in the O/W/O (oil/water/oil) system using a U-shaped cell. The transport rate and the preferred enantiomer of the esters were substantially affected by pH, temperature, and species of albumin. Among five serum albumins studied, bovine serum albumin (BSA) showed the largest rate constant and rat serum albumin (RSA) manifested the highest enantioselectivity. Regarding enantiomer selectivity in transport overall, it is anticipated that the ester uptake step plays an important role for BSA, whereas the ester release is the key step for RSA. PMID- 10368927 TI - Isolating the contributions of familiarity and source information to item recognition: a time course analysis. AB - Recognition memory may be mediated by the retrieval of distinct types of information, notably, a general assessment of familiarity and the recovery of specific source information. A response-signal speed-accuracy trade-off variant of an exclusion procedure was used to isolate the retrieval time course for familiarity and source information. In 2 experiments, participants studied spoken and read lists (with various numbers of presentations) and then performed an exclusion task, judging an item as old only if it was in the heard list. Dual process fits of the time course data indicated that familiarity information typically is retrieved before source information. The implications that these data have for models of recognition, including dual-process and global memory models, are discussed. PMID- 10368928 TI - Memory representation of alphabetic position and interval information. AB - The authors conducted 3 sets of experiments. In the 1st set of experiments, participants made alphabetic position estimations. In the 2nd set, participants made interletter distance estimations. In the 3rd set, they made comparative judgments of the alphabetic order of a pair of letters. The results showed that participants had highly accurate ordinal level information about the alphabet in memory but that interval level information was systematically distorted. In addition, alphabetic serial information was found to be used in 2 distinct modes in memory, depending on whether the representation could be contained within the span of immediate memory. PMID- 10368929 TI - Dual-coding, context-availability, and concreteness effects in sentence comprehension: an electrophysiological investigation. AB - Event-related potentials were recorded in 2 experiments while participants read sentences in a word-by-word congruency judgment task. Sentence final words were either congruent, semantically anomalous (Experiments 1 and 2), or neutral (Experiment 2) with respect to sentence context. Half of all final words referred to concrete and half to abstract concepts. A different scalp distribution of the N400 to concrete and abstract final words was found for anomalous and neutral, but not congruent sentences. Although the interaction of context and concreteness is consistent with the context-availability model, the differential scalp distribution of effects for concrete and abstract words, as well as larger context effects for concrete words, was interpreted as being more consistent with an extended dual-code account of semantic processing. PMID- 10368930 TI - Inoculative freezing by environmental ice nuclei in the freeze-tolerant wood frog, Rana sylvatica. AB - Efficacy of inoculative freezing by ice nuclei in a simulated winter environment was studied in the wood frog (Rana sylvatica), a freeze-tolerant species that overwinters on the forest floor beneath organic detritus. Adult frogs were confined to plastic canisters and cooled to -2 degrees C over 24 hr with their ventral skin in contact with substrate (humic soil hydrated to 40, 10, or 5%, or soil/peat mixture hydrated to 20 or 10%, w/w), or their dorsal skin in contact with damp leaf mould. Whereas only 20% of control frogs cooled in dry, plastic canisters froze, freezing occurred in nearly all (98%) frogs contacting soil or leaf mould. Inoculation was briefly delayed in frogs exposed to drier substrates. Frogs exposed to an unfreezable substrate (humic soil, 5% moisture) themselves froze, apparently due to the action of constituent nuclei which commonly occur in natural materials. Although the surface over which inoculation can occur is greater in larger frogs, inoculation susceptibility was not correlated with body mass in our frogs (mean +/- SE body mass = 14.0 +/- 0.2 g; range, 9.8-17.8 g). We conclude that the high susceptibility to inoculative freezing in R. sylvatica, which is conferred by its moist, highly permeable integument, promotes freeze tolerance by ensuring that inoculation commences at relatively high temperatures. PMID- 10368931 TI - Low molecular weight acid phosphatase/phosphotyrosyl protein phosphatase in the developing chick brain: partial characterization and levels during development. AB - Low molecular weight acid phosphatase/phosphotyrosyl protein phosphatase is largely expressed in chick brain tissue during development. The enzyme was purified from brain extract prepared from 19-day-old chick embryos and from adult chickens using ammonium sulfate fractionation, gel filtration on Sephadex G-75 and two DEAE-Cellulose ion-exchange chromatography steps. The purified enzymes from embryo and adult chick brains show identical molecular weight values (about 18-20 kDa) and biochemical and structural properties such as substrate specificity, sensitivity to inhibitors, and number of free reactive sulphydryl groups. These data suggest that they are the same enzyme protein. Although the total acid phosphatase activity does not change appreciably during development, the activity associated with the low molecular weight acid phosphatase/phosphotyrosyl protein phosphatase markedly increases after birth and reaches the adult values within the first week of life. Taken together, our results suggest an involvement of the low molecular weight acid phosphatase/phosphotyrosyl protein phosphatase in postnatal development and maturation of chick brain tissue. The variations in tyrosine phosphorylation profile of chick brain polypeptides analyzed by Western blotting at the same developmental stages are also reported. PMID- 10368932 TI - Influence of torpor on milk protein composition and secretion in lactating bats. AB - In the pipistrelle bat (Pipistrellus pipistrellus), the metabolic load of lactation is not met to any significant extent by increased food intake or mobilization of body reserves, and aerial foraging accounts for most of the animal's energy expenditure even during lactation. Energy conservation must, therefore, play a critical role in maintaining lactation. The principal mechanism for energy conservation appears to be the bat's ability to enter torpor, but this may itself interrupt milk synthesis and secretion unless the pipistrelle mammary gland is adapted to counteract its effect. The effect of torpor on mammary tissue function was studied in mammary tissue explant cultures prepared in weeks 1-3 of lactation, when milk water yield was 0.20, 0.88, and 0.30 mL/d respectively. Protein synthesis measured by incorporation of radiolabeled amino acids was 44% lower (P < 0.001) in bat tissue explants cultured at ambient temperature (22 degrees C) compared with 37 degrees C. The reduction was similar to that observed in mouse mammary tissue (57%) and was unaffected by stage of lactation. Analysis of explant protein after [35S]methionine labelling showed the majority of proteins synthesised in culture to be milk proteins; it also demonstrated that the decrease in protein synthesis at ambient temperature was a general phenomenon: synthesis of both secretory and intracellular mammary proteins was reduced at the lower culture temperature. The results suggest that bat mammary tissue has no mechanism to counteract the effect of reduced body temperature and that periods of lactational torpor are likely to cause a pronounced diurnal variation in the rate of milk secretion. PMID- 10368933 TI - Physiological changes in the brushtail possum (Trichosurus vulpecula) following relocation from Armidale to Brisbane, Australia. AB - To determine the effect of relocation on the health of possums the body weights and hormone and immune responses of six male and nine female brushtail possums were monitored for 20 weeks following transfer from the environs of Armidale into enclosures in Brisbane. Over the first 6 weeks of captivity, male possums lost 11.0% of their original body weight and females lost 16.8%. The mean concentrations of plasma cortisol in the male and female possums were 14.5 and 29.4 ng/mL, respectively, and did not change over the 20-week period. Male and female possums displayed a similar pattern of thyroxine secretion over the 20 weeks, with low concentrations up to week seven (2.1 and 2.7 ng/mL, respectively) increasing to 6.9 and 5.8 ng/mL in weeks 7-12 (P < 0.005). This increase in the concentration of thyroxine corresponded with the increase in body weight. The number of white blood cells (WBCs) and the percentage of neutrophils increased from the capture to week 6-10. However, during the last 10 weeks of captivity the number of WBCs and the percentage of neutrophils decreased, indicating recovery of the immune system. This was in accord with the proliferative response of lymphocytes to the T cell mitogen PHA that increased from weeks 11-15 to weeks 16 20 in both male and female possums. The results above suggest that the Armidale possums, like the Brisbane possums, were stressed following their relocation; however, their immune systems were able to gradually recover as they adjusted to their new environment in Brisbane. The death rate of pouch young and of adult female possums after relocation was considerably higher in the Armidale possums compared to Brisbane possums. The mortality rate of Brisbane possums over the first 20 weeks of captivity was 8.3% and 19.6% for male and female possums, respectively, and for Armidale possums 16.6% and 47.1%, respectively. The possums transferred from the environs of Armidale into captivity in Brisbane were under greater stress than possums captured in Brisbane and placed in captivity in Brisbane. PMID- 10368934 TI - The sea within us. PMID- 10368935 TI - Use of developmental marker genes to define temporal and spatial patterns of differentiation during embryoid body formation. AB - Mouse embryonic stem cells are pluripotent cells that are derived from the inner cell mass of blastocysts. When induced to synchronously enter a program of differentiation in vitro, they form embryoid bodies that contain cells of the mesodermal, hematopoietic, endothelial, muscle, and neuronal lineages. Here, we used a panel of marker genes with early expression within the germ layers (oct-3, Brachyury T, Fgf-5, nodal, and GATA-4) or a variety of lineages (flk-1, Nkx-2.5, EKLF, and Msx3) to determine how progressive differentiation of embryoid bodies in culture correlated with early postimplantation development of mouse embryos. Using RNA in situ hybridization, we found that the temporal and spatial relationships existing between these marker genes in vivo were maintained also in vitro. Studying the onset of marker gene expression allowed us also to determine the time course of differentiation during the formation of embryoid bodies. Thus, stages equivalent to embryogenesis between implantation and the beginning of gastrulation (4.5-6.5 d.p.c.) occur within the first two days of embryoid body differentiation. Between days 3 and 5, embryoid bodies contain cell lineages found in embryos during gastrulation at 6.5 to 7.0 d.p.c., and after day 6 in culture, embryoid bodies are equivalent to early organogenesis-stage embryos (7.5 d.p.c.). In addition, we demonstrate that the panel of developmental markers can be applied in a screen for stage- or lineage-specific genes. Reporter gene expression from entrapment vector insertions can be co-localized with expression of specific markers within the same cell during embryoid body formation as well as during embryogenesis. Our results thus demonstrate the power of embryoid body formation as an in vitro model system to study early lineage determination and organogenesis in mammals, and indicate that they will prove to be useful tools for identifying developmental genes whose expression is restricted to particular lineages. PMID- 10368936 TI - Effects of insulin-like growth factor 1 and testosterone on the proliferation of antlerogenic cells in vitro. AB - Androgen hormones and growth factors are implicated in pedicle formation and antler transformation in deer. The potential to form a pedicle and an antler is only found in the antlerogenic periosteum (AP) overlying the presumptive antler growth region. Histological studies (Li and Suttie, '94) showed that AP consists of an inner cellular layer and an outer fibrous layer. Pedicle and antler are mainly derived from the cellular layer cells of the AP. Ossification takes place in four stages: intramembranous (IMO), transitional (OPC), pedicle endochondral (pECO) and antler endochondral (aECO). However, the precise mechanism whereby androgen hormones and growth factors control pedicle and antler formation is unknown. The aim of this study was to use cell culture techniques to investigate how testosterone and IGF1 affects the proliferation of antlerogenic cells from the four ossification stages of pedicle/antler in vitro. The results showed that in serum-free medium IGF1 stimulated the proliferation of antlerogenic cells from all four ossification stages in a dose-dependent manner. In contrast, testosterone alone did not show any mitogenic effects on these antlerogenic cells. However, in the presence of IGF1, testosterone increased proliferation of the antlerogenic cells from the IMO and the OPC stages (pedicle tissue), and reduced proliferation of the antlerogenic cells from transformation point (TP) and aECO stages (antler tissue). Therefore, the results from the present in vitro study support the in vivo findings that androgen hormones stimulate pedicle formation but inhibit antler growth. The change in the mitogenic effects of testosterone on antlerogenic cells from positive to negative occurs approximately at the change in ossification type from OPC to pECO. Therefore, these results reinforce the hypothesis that the transformation from a pedicle to an antler takes place at the time when the ossification type changes from OPC to pECO rather than at the time when the pedicle grows to its full species-specific height. PMID- 10368937 TI - Daily scheduling of the golden spiny mouse under photoperiodic and social cues. AB - A most important function of the circadian system is to ensure that behaviors and metabolism are appropriately timed with respect to the light/dark cycle and photoperiod. Ecological constraints can perturb the daily schedules; would they also impair photoperiodic adaptations? A natural model exists in the golden spiny mouse (Acomys russatus), which is nocturnal, but driven into diurnal activity when sharing the habitat with its congener, A. cahirinus. We show here that the presence of A. cahirinus alters the diurnal rhythms of body temperature and urine volume, delays excretion of the major melatonin metabolite, 6-sulfatoxymelatonin (6-SMT), and increases 2-deoxyglucose uptake by the suprachiasmatic nuclei in A. russatus. Nevertheless, a clear photoperiod effect on urine volume and 6-SMT rhythms was observed. These results indicate that the circadian system can adapt to major changes in daily scheduling without impairing daylength measurement, and consequently seasonal adaptation. PMID- 10368938 TI - Development of an efficient method to produce uniformly haploid parthenogenones. AB - Bovine ovaries (paired by cow) were obtained from a local abattoir and cumulus oocyte complexes were aspirated within six hours of slaughter. Two methods for activation [(1) calcium ionophore (ionomycin) alone (n = 191); and (2) ionomycin followed by the protein synthesis inhibitor cycloheximide (n = 207)] were evaluated for production of bovine parthenogenones. Activation with ionomycin alone resulted in a development rate of 33%, while activation with ionomycin and cycloheximide sequentially resulted in a development rate to two-cell stage of 49%. A procedure was developed to expedite accurate evaluation of activated oocytes for uniformly haploid development. Uniformly haploid parthenogenones that cleaved at least once in four days of in vitro culture were individually prepared for genetic analysis. Three techniques: (1) phosphate buffered saline; (2) TL HEPES with 0.2% ovine serum albumin; and (3) TL-HEPES with 0.2% polyvinyl pyrrolidone were compared to harvest parthenogenones for genetic analysis. The only effective method that did not create spurious results during later genetic analysis was TL-HEPES with 0.2% polyvinyl pyrrolidone. Based on the results of this study, we estimate that an average of 5-7 uniformly haploid bovine parthenogenones can be realized from each donor (using pairs of ovaries). These parthenogenones, when maintained as family units, will be valuable for accomplishment of female-specific genetic linkage analysis. PMID- 10368939 TI - Age and the allocation of attention across the time course of word recognition. AB - Younger and older adults performed lexical decisions on ambiguous words, unambiguous words, and pseudowords, and simultaneously responded to an auditory probe presented at stimulus onset asynchronies (SOAs) of 90, 180, or 270 ms. For both age groups, lexical decisions and probe responses were faster for ambiguous words than for unambiguous words, and slowest for pseudowords. For the older adults, but not the younger adults, lexical decisions were slower when the probe was presented (the dual-task condition), compared with a control condition in which the lexical decision was performed alone. The older participants also showed slower tone-detection responses in the dual-task condition than when the tone was presented alone. For all participants, proportional tone-detection times (compared with tones in isolation) decreased with increasing SOA, but this decrease was less pronounced in the older group. Finally, the time between responses in the dual-task condition was longer for older than for younger adults. The results indicate that word meaning influences the allocation of attention similarly for younger and older adults, but that older adults suffer a cost and become disproportionately slower in processes related to response coordination and output. PMID- 10368940 TI - Sociodemographic characteristics and drinking status as predictors of older women's health. AB - As part of a U.S. national survey of women's drinking and life experiences, the authors used responses from a subsample (n = 245) of women aged 55-90 years (M = 65.8 years) to examine the relationship of sociodemographic characteristics (income, marital status, and occupational status) and drinking status to several health outcomes (self-perceived general health, depression, sexual satisfaction, and sexual dysfunction). In all analyses, the authors controlled for respondent age. Results indicated that higher household income predicted greater lifetime and current sexual satisfaction with a partner as well as higher general health ratings. Women drinkers also reported better general health than did abstainers. An interaction between marital status (married or cohabitating vs. nonmarried) and employment status (employed vs. nonemployed) was a predictor of general health ratings. The authors found significant contrasts among the 4 groups when they controlled for age, income, and drinking status: (a) Among the employed respondents, the nonmarried women reported better general health than did the married women; and (b) among nonmarried respondents, the employed women reported better general health than did the nonemployed women. PMID- 10368941 TI - Locus of control and the age difference in free recall from episodic memory. AB - The authors investigated the relation of locus of control (LOC) to age differences in free-recall memory performance. Older and younger participants completed P. C. Duttweiler's (1984) Internal Control Index (ICI) and subsequently performed free-recall memory tasks. Compared with the younger participants, the older participants exhibited poorer recall with more intrusions and uncorrected repetition errors as well as reduced categorical clustering. For the older participants with less internal LOC, recall proportion and item-pair associative recall clustering were lower than for the older participants with more internal LOC. By contrast, the younger participants did not exhibit any LOC effects in their recall performance. The results suggest that a differential memory organization deficit may underlie the age differences in free recall among individuals varying in LOC when they are performing an intentional learning task. This deficit is discussed in terms of a reduced-inhibition account of cognitive aging. PMID- 10368942 TI - Human behavioral momentum in a sample of older adults. AB - Behavioral momentum, the persistence of behavior under altered environmental contingencies, is derived from Newtonian physics and operant psychology. It has relevance to behavior analysis in terms of shaping strong behaviors and ensuring effective relapse prevention strategies in behavior modification and therapy. The authors investigated whether changing the operant schedule contingencies affects the responses of older humans to different stimuli when reinforcement density is systematically manipulated. Fifteen older adults participated in a computer study in which each of 2 keys in a baseline condition was associated with the same schedule of reinforcement and multiple variable intervals; the only difference was that 1 reinforcer was 10 times larger than the other. After 6 sessions, the authors changed the contingency schedule to either an extinction condition, a variable-time schedule, or a different variable-interval schedule, to assess how participants' responses persisted when reinforcement contingencies were systematically changed. The results were consistent with the predictions of behavioral momentum. The participants not only biased their responses in favor of the more densely reinforcing key, but when contingencies changed, they showed significantly biased responses. Results supported the conclusion that healthy older adults allocate their behaviors in a manner very sensitive to training stimuli conditions; consistent with the basic principles of behavioral momentum, they show a degree of resistance to change in their behaviors when the behavioral contingencies are altered. PMID- 10368943 TI - Regular versus randomized sentences, nouns versus prepositions, and assimilation in salience. AB - The authors' research supports an alternative to the theory that organization improves retention by producing fewer elements for processing. College students' recall of randomized words was better for nouns than for prepositions. Regular sentences improved recall, but this improvement was less for nouns than for prepositions. According to the alternative theory, a part assimilates (increases in similarity) to its high-in-salience organization-produced group, thus increases in salience, and hence is retained better. The high-in-salience group for a regular sentence should be the sentence's meaning. Because the nouns were recalled better, they were more similar in salience to the regular sentences' meanings than were the prepositions. Assimilation should be minimal when parts are very similar. Therefore, the nouns' assimilation in salience should have been less than the prepositions', explaining the nouns' less improvement in recall. PMID- 10368944 TI - Influence of the specific immune response on some consistent murine behaviors. AB - The author's goal was to discover if the generation and maintenance of the specific immune response resulted in alterations of reliable behaviors (i.e., behaviors correlated over time). The behaviors (ambulation, rearing, and interaction with a conspecific) of CD1 male mice were measured in a small open field, and several days later, the mice were immunized with antigens (either splenocytes from C57BL/6 mice or a mixture of sheep erythrocytes and goat serum). The same behaviors were recorded again some hours, or some days, after immunization. Immunizations and behavioral measurements were repeated at various intervals. Blood levels of antibodies to the antigens were measured 6 days after immunization. The recorded behaviors were consistent (according to Kendall coefficient of concordance). The mice mounted antibody responses to the antigens, yet no behavioral changes were apparent during the response. On the contrary, a single injection of E. coli lipopolysaccharide decreased ambulation and rearing. It is proposed that in healthy mice kept in normal conditions, the specific immune response may be unrelated to reliable behavioral changes. PMID- 10368945 TI - Investigation of cell culture media infected with viruses by pyrolysis mass spectrometry: implications for bioaerosol detection. AB - Mass spectrometry coupled with a pyrolysis inlet system was used to investigate media from cell cultures infected with viruses. Cell culture media is an intricate mixture of numerous chemical constituents and cells that collectively produce complicated mass spectra. Cholesterol and free fatty acids were identified and attributed to lipid sources in the media (blood serum supplement and plasma membranes of host cells). These lipid moieties could be utilized as signature markers for rapidly detecting the cell culture media. Viruses are intracellular parasites and are dependent upon host cells in order to exist. Therefore, it is highly probable that significant quantities of media needed to grow and maintain viable host cells would be present if a viral agent were disseminated as an aerosol into the environment. Cholesterol was also detected from a purified virus sample, further substantiating its use as a target compound for detection. Implications of this research for detection of viral bioaerosols, using a field-portable pyrolysis mass spectrometer, is described. PMID- 10368946 TI - Observation of gel-induced protein modifications in sodium dodecylsulfate [corrected] polyacrylamide gel electrophoresis and its implications for accurate molecular weight determination of gel-separated proteins by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. AB - Matrix-assisted laser desorption ionization (MALDI) time-of-flight mass spectrometry (TOFMS) can potentially provide accurate molecular weight information of proteins separated by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). Several issues related to resolution and accuracy of molecular weight measurement are investigated by using a time-lag focusing MALDI TOF mass spectrometer. The effects of the gel components SDS, glycerol, and tris buffer on the mass spectral signals are studied systematically. Glycerol and tris buffer are shown to have little or no effect on resolution and mass accuracy, whereas SDS degrades sensitivity, resolution, and mass accuracy even at low concentrations. A simple and fast gel extraction technique is presented which is capable of detecting proteins loaded at the low-picomole level on the gel. The sample preparation procedure used in this work appears to remove most of SDS from the gel, thereby reducing the peak broadening effect caused by SDS and resulting in high resolution and accurate measurement of proteins. However, for proteins containing cysteines, the molecular ions are composed of a distribution of acrylamide-protein adducts likely formed by reaction with unpolymerized acrylamide in the gel during the gel separation process. The implications of gel induced protein modifications on the accurate molecular weight measurement of gel separated proteins are discussed. PMID- 10368947 TI - Identification of single stranded regions of DNA by enzymatic digestion with matrix-assisted laser desorption/ionization analysis. AB - Elucidating structure function relationships of DNA in cellular processes requires fast, reliable methods that can be applied to picomole amounts of sample. Higher order structure can be inferred by distinguishing paired and unpaired regions. It is shown here that enzymatic digestion coupled with product analysis by matrix-assisted laser desorption ionization (MALDI) is able to identify unpaired bases within structured DNA regions. The method is demonstrated with DNA duplexes having a five nucleotide mismatch as a 5' overhang, a 3' overhang, and an internal loop. Exo- and endonuclease digestions are performed under solution conditions (temperature, annealing, and enzyme buffers) which promote base pairing and specific enzyme activity. For each type of mismatch, the length and sequence of the single stranded region can be inferred from MALDI spectra taken as a function of digestion time. PMID- 10368948 TI - Mass spectrometric and tandem mass spectrometric behavior of nitrocatechol glucuronides: a comparison of atmospheric pressure chemical ionization and electrospray ionization. AB - The mass spectrometric (MS) and tandem mass spectrometric (MS/MS) behavior of six nitrocatechol-type glucuronides using atmospheric pressure chemical ionization (APCI) and electrospray ionization (ESI) was systematically studied, and the effect of operation parameters on the fragmentations are presented. The positive ion APCI- and ESI-MS spectra showed an intense protonated molecule and the respective negative ion spectra a deprotonated molecule with minimal fragmentation. The main fragment ions in the MS/MS spectra of the protonated and deprotonated molecules were [M + H - Glu]+ and [M - H - Glu]-, respectively, formed by the loss of the glucuronide moiety. The measured limits of detection indicated that ESI is a significantly more efficient ionization method than APCI in the negative and positive ion modes for the compounds studied. MS/MS was found to be less sensitive, but more reliable and simple than MS due to the absence of chemical noise. PMID- 10368949 TI - Hydroiodic acid attachment kinetics as a chemical probe of gaseous protein ion structure: bovine pancreatic trypsin inhibitor. AB - The kinetics of attachment of hydroiodic acid (HI) to the (M + 6H)6+ ions of native and reduced forms of bovine pancreatic trypsin inhibitor (BPTI) in the quadrupole ion trap environment are reported. Distinctly nonlinear (pseudo first order) reaction kinetics are observed for reaction of the native ions, indicating two or more noninterconverting structures in the parent ion population. The reduced form, on the other hand, shows very nearly linear reaction kinetics. Both forms of the parent ion attach a maximum of five molecules of hydroiodic acid. This number is expected based on the amino acid composition of the protein. There is a total of 11 strongly basic sites in the protein (i.e., six arginines, four lysines, and one N-terminus). An ion with protons occupying six of the basic sites has five available for hydroiodic acid attachment. The kinetics of successive attachment of HI to the native and reduced forms of BPTI also differ, particularly for the addition of the fourth and fifth HI molecules. A very simple kinetic model describes the behavior of the reduced form reasonably well, suggesting that all of the neutral basic sites in the reduced BPTI ions have roughly equal reactivity. However, the behavior of the native ion is not well described by this simple model. The results are discussed within the context of differences in the three-dimensional structures of the ions that result from the presence or absence of the three disulfide linkages found in native BPTI. The HI reaction kinetics appears to have potential as a chemical probe of protein ion three-dimensional structure in the gas phase. Hydroiodic acid attachment chemistry is significantly different from other chemistries used to probe three dimensional structure and hence, promises to yield complementary information. PMID- 10368950 TI - Evolution of DNA base composition under no-strand-bias conditions when the substitution rates are not constant. AB - The evolution of DNA base composition evolution is simplified to a six-parameter model when there are no strand biases for mutation and selection. We analyzed the dynamics of this model with special attention to the influence of a change in substitution rates. The G + C content of the DNA sequence tends to an equilibrium value that is controlled by four parameters of the model. When the substitution rates are not constant, the G + C equilibrium position is not constant. The DNA sequence base frequencies always tend to a state in which A = T and G = C within a strand, regardless of substitution rates. This is true even when the substitution rates are not constant over time. This provides a simple way of rejecting the model from inspection of present-day DNA base composition. PMID- 10368951 TI - Nucleotide variation at the runt locus in Drosophila melanogaster and Drosophila simulans. AB - Intra- and interspecific nucleotide variation for the major developmental gene runt in Drosophila was studied in D. melanogaster and D. simulans. The 1.5-kb protein-coding region and the 0.4-kb intron of the runt gene were sequenced for 11 alleles in each species. The D. melanogaster alleles originated from east Africa. Estimated parameters of intraspecific variation in D. melanogaster (exons: theta = 0.018, pi = 0.018; intron: theta = 0.014, pi = 0.014) and D. simulans (exons: theta = 0.007, pi = 0.005; intron: theta = 0.008, pi = 0.005) were below average for other X-linked genes, while divergence between species (exons: D = 0.094; intron: D = 0.069) fell within the normal range for both silent and replacement changes. This estimate for runt, along with published values for three other genes in regions of normal recombination, show east African D. melanogaster to be roughly twice as polymorphic as D. simulans. The majority of nucleotide variation, silent and replacement, in both species was found to be selectively neutral using various statistical tests (HKA, McDonald Kreitman, Tajima, and Fu and Li tests). Monte Carlo simulations of the coalescent process significantly rejected a Wright-Fisher model with respect to an amino acid polymorphism and the distribution of polymorphic sites among the D. simulans lines. This indicated an old lineage and may reflect ancestral population substructuring in D. simulans. PMID- 10368952 TI - The novel mitochondrial gene arrangement of the cattle tick, Boophilus microplus: fivefold tandem repetition of a coding region. AB - We sequenced across all of the gene boundaries in the mitochondrial genome of the cattle tick, Boophilus microplus, to determine the arrangement of its genes. The mtDNA of B. microplus has a coding region, composed of tRNA(Glu) and 60 bp of the 3' end of ND1, that is repeated five times. Boophilus microplus is the first coelomate animal known to have more than two copies of a coding sequence. The mitochondrial genome of B. microplus has other unusual features, including (1) reduced T arms in tRNAs, (2) an AT bias in codon use, (3) two control regions that have evolved in concert, (4) three gene rearrangements, and (5) a stem-loop between tRNA(Gln) and tRNA(Phe). The short T arms and small control regions (CRs) of B. microplus and other ticks suggest strong selection for small genomes. Imprecise termination of replication beyond its origin, which can account for the evolution of tandem repeats of coding regions in other mitochondrial genomes, cannot explain the evolution of the fivefold repeated sequence in the mitochondrial genome of B. microplus. Instead, slipped-strand mispairing or recombination are the most plausible explanations for the evolution of these tandem repeats. PMID- 10368953 TI - The influence of recombination on the population structure and evolution of the human pathogen Neisseria meningitidis. AB - The extent to which recombination disrupts the bifurcating treelike phylogeny and clonal structure imposed by binary fission on bacterial populations remains contentious. Here, we address this question with a study of nucleotide sequence data from 107 isolates of the human pathogen Neisseria meningitidis. Gene fragments from 12 house-keeping loci distributed around the meningococcal chromosome were analyzed, showing that (1) identical alleles are disseminated among genetically diverse isolates, with no evidence for linkage disequilibrium; (2) different loci give distinct and incongruent phylogenetic trees; and (3) allele sequences are incompatible with a bifurcating treelike phylogeny at all loci. These observations are consistent with the hypothesis that meningococcal populations comprise organisms assembled from a common gene pool, with alleles and allele fragments spreading independently, together with the occasional importation of genetic material from other species. Further, they support the view that recombination is an important genetic mechanism in the generation new meningococcal clones and alleles. Consequently, for anything other than the short term evolution of this species, a bifurcating treelike phylogeny is not an appropriate model. PMID- 10368954 TI - Genomic and evolutionary analysis of Feilai, a diverse family of highly reiterated SINEs in the yellow fever mosquito, Aedes aegypti. AB - Five short interspersed repetitive elements (SINEs) were found fortuitously in the introns of a steroid hormone receptor AaHR3-2 gene of the yellow fever mosquito, Aedes aegypti, constituting a novel family of tRNA-related SINEs named Feilai. In addition, nine other Feilai elements were found in currently available sequences in Ae. aegypti, six of which were also near genes. Approximately 5.9 x 10(4) copies of Feilai were present in Ae. aegypti, equivalent to 2% of the entire genome. An additional 35 Feilai elements were isolated from a genomic library. Of the total 49 Feilai elements, 20 were full-length. Sequence comparisons and phylogenetic analyses of the full-length elements strongly suggest that there are at least two subfamilies within the Feilai family. There is a high degree of conservation within the two subfamilies. However, sequence divergence between the subfamilies, along with the presence of highly degenerate Feilai elements, suggests that Feilai is likely a diverse family of SINEs that has existed in Ae. aegypti for a long time. Many Feilai elements were closely associated with other transposons, especially with fragments of non-LTR retrotransposons and miniature inverted-repeat transposable elements. The 500-bp sequences immediately flanking a Feilai element were highly A + T-rich, which is consistent with the fact that no Feilai has been found in the coding regions of genes. It is likely that the highly reiterated and interspersed Feilai elements are partially responsible for the pattern of short-period interspersion of the Ae. aegypti genome. The evolutionary relationship between Feilai and the Ae. aegypti genome is likely complex. PMID- 10368955 TI - Networks and groups within the genus Neisseria: analysis of argF, recA, rho, and 16S rRNA sequences from human Neisseria species. AB - To understand the pattern of nucleotide sequence variation among bacteria that frequently exchange chromosomal genes, we analyzed sequences of the recA, argF, and rho genes, as well as part of the small-subunit (16S) rRNA gene, from about 50 isolates of human commensal Neisseria species and the pathogenic N. meningitidis and N. gonorrhoeae. Almost all isolates of these species could be assigned to five phylogenetic groups that are found for all genes examined and generally are supported by high bootstrap values. In contrast, the phylogenetic relationships among groups varied according to the gene analyzed with notable incongruences involving N. cinerea and N. lactamica. Further analysis using split decomposition showed that for each gene, including 16S rRNA, the patterns of sequence divergence within N. meningitidis and closely related species were inconsistent with a bifurcating treelike phylogeny and better represented by an interconnected network. These data indicate that the human commensal Neisseria species can be separated into discrete groups of related species but that the relationships both within and among these groups, including those reconstructed using 16S rRNA, have been distorted by interspecies recombination events. PMID- 10368956 TI - Complete mitochondrial DNA sequences of the green turtle and blue-tailed mole skink: statistical evidence for archosaurian affinity of turtles. AB - Turtles have highly specialized morphological characteristics, and their phylogenetic position has been under intensive debate. Previous molecular studies have not established a consistent and statistically well supported conclusion on this issue. In order to address this, complete mitochondrial DNA sequences were determined for the green turtle and the blue-tailed mole skink. These genomes possess an organization of genes which is typical of most other vertebrates, such as placental mammals, a frog, and bony fishes, but distinct from organizations of alligators and snakes. Molecular evolutionary rates of mitochondrial protein sequences appear to vary considerably among major reptilian lineages, with relatively rapid rates for snake and crocodilian lineages but slow rates for turtle and lizard lineages. In spite of this rate heterogeneity, phylogenetic analyses using amino acid sequences of 12 mitochondrial proteins reliably established the Archosauria (birds and crocodilians) and Lepidosauria (lizards and snakes) clades postulated from previous morphological studies. The phylogenetic analyses further suggested that turtles are a sister group of the archosaurs, and this untraditional relationship was provided with strong statistical evidence by both the bootstrap and the Kishino-Hasegawa tests. This is the first statistically significant molecular phylogeny on the placement of turtles relative to the archosaurs and lepidosaurs. It is therefore likely that turtles originated from a Permian-Triassic archosauromorph ancestor with two pairs of temporal fenestrae behind the skull orbit that were subsequently lost. The traditional classification of turtles in the Anapsida may thus need to be reconsidered. PMID- 10368957 TI - The age and evolution of non-LTR retrotransposable elements. AB - A comprehensive phylogenetic analysis was conducted of non-long-terminal-repeat (non-LTR) retrotransposons based on an extended sequence alignment of their reverse transcriptase (RT) domain. The 440 amino acid positions used included a region proposed to be similar to the "thumb" of the right-handed RT structure found in retroviruses. All identified non-LTR elements could be grouped into 11 distinct clades. Using the rates of sequence change derived from studies of the vertical inheritance of R1 and R2 elements in arthropods as a comparison, we found no evidence for the horizontal transmission of non-LTR elements. Assuming vertical descent, the phylogeny suggested that non-LTR elements are as old as eukaryotes, with each of the 11 clades dating back to the Precambrian era. The analysis enabled us to propose a simple chronology for the acquisition of different enzymatic domains in the evolution of the non-LTR class of retrotransposons. The first non-LTR elements were sequence specific by virtue of a restriction-enzyme-like endonuclease located downstream of the RT domain. Evolving from this original group were elements (eight clades) that acquired an apurinic-apyrimidic endonuclease-like domain upstream of the RT domain. Finally, four of these clades have inherited an RNase H domain downstream of the RT domain. The phylogenies of the AP endonuclease and RNase H domains were also determined for this report and are consistent with the monophyletic acquisition of these domains. These studies represent the most comprehensive effort to date to trace the evolution of a major class of transposable elements. PMID- 10368958 TI - Generalized neighbor-joining: more reliable phylogenetic tree reconstruction. AB - We have developed a phylogenetic tree reconstruction method that detects and reports multiple topologically distant low-cost solutions. Our method is a generalization of the neighbor-joining method of Saitou and Nei and affords a more thorough sampling of the solution space by keeping track of multiple partial solutions during its execution. The scope of the solution space sampling is controlled by a pair of user-specified parameters--the total number of alternate solutions and the number of alternate solutions that are randomly selected- effecting a smooth trade-off between run time and solution quality and diversity. This method can discover topologically distinct low-cost solutions. In tests on biological and synthetic data sets using either the least-squares distance or minimum-evolution criterion, the method consistently performed as well as, or better than, both the neighbor-joining heuristic and the PHYLIP implementation of the Fitch-Margoliash distance measure. In addition, the method identified alternative tree topologies with costs within 1% or 2% of the best, but with topological distances of 9 or more partitions from the best solution (16 taxa); with 32 taxa, topologies were obtained 17 (least-squares) and 22 (minimum evolution) partitions from the best topology when 200 partial solutions were retained. Thus, the method can find lower-cost tree topologies and near-best tree topologies that are significantly different from the best topology. PMID- 10368959 TI - Archaea sister group of Bacteria? Indications from tree reconstruction artifacts in ancient phylogenies. AB - The 54-kDa signal recognition particle and the receptor SR alpha, two proteins involved in the cotranslational translocation of proteins, are paralogs. They originate from a gene duplication that occurred prior to the last universal common ancestor, allowing one to root the universal tree of life. Phylogenetic analysis using standard methods supports the generally accepted cluster of Archaea and Eucarya. However, a new method increasing the signal-to-noise ratio strongly suggests that this result is due to a long-branch attraction artifact, with the Bacteria evolving fastest. In fact, the Archaea/Eucarya sisterhood is recovered only by the fast-evolving positions. In contrast, the most slowly evolving positions, which are the most likely to retain the ancient phylogenetic signal, support the monophyly of prokaryotes. Such a eukaryotic rooting provides a simple explanation for the high similarity of Archaea and Bacteria observed in complete-genome analysis, and should prompt a reconsideration of current views on the origin of eukaryotes. PMID- 10368960 TI - Molecular evolution of glutamate receptors: a primitive signaling mechanism that existed before plants and animals diverged. AB - We performed a genealogical analysis of the ionotropic glutamate receptor (iGluR) gene family, which includes the animal iGluRs and the newly isolated glutamate receptor-like genes (GLR) of plants discovered in Arabidopsis. Distance measures firmly placed the plant GLR genes within the iGluR clade as opposed to other ion channel clades and indicated that iGluRs may be a primitive signaling mechanism that predated the divergence of animals and plants. Moreover, phylogenetic analyses using both parsimony and neighbor joining indicated that the divergence of animal iGluRs and plant GLR genes predated the divergence of iGluR subtypes (NMDA vs. AMPA/KA) in animals. By estimating the congruence of the various glutamate receptor gene regions, we showed that the different functional domains, including the two ligand-binding domains and the transmembrane regions, have coevolved, suggesting that they assembled together before plants and animals diverged. Based on residue conservation and divergence as well as positions of residues with respect to functional domains of iGluR proteins, we attempted to examine structure-function relationships. This analysis defined M3 as the most highly conserved transmembrane domain and identified potential functionally important conserved residues whose function can be examined in future studies. PMID- 10368961 TI - Rapid evolution of fertilization selectivity and lysin cDNA sequences in teguline gastropods. AB - Proteins mediating intercellular recognition face opposing selective forces as they evolve: purifying selection to maintain function, and diversifying selection to alter specificity. Lysin is a 16-kDa protein which enables sperm of free spawning marine snails to make a hole in the vitelline layer (VE) surrounding conspecific eggs. Previous work on abalone (Haliotis spp.) has shown that positive selection promotes rapid interspecific divergence of lysin. Here, we present data on the specificity of VE dissolution by four species of teguline gastropods, along with lysin cDNA sequences. The teguline and abalone lineages diverged over 250 MYA. As in abalone, VE dissolution by lysin in tegulines is species-selective, and positive selection promotes rapid interspecific divergence over the entire mature protein. Nonsynonymous substitution rates, calculated using a mtCOI molecular clock calibrated by two Tegula species separated by the Isthmus of Panama, are high (> 25 substitutions per site per 10(9) years). However, the extensive replacements in teguline lysins are overwhelmingly conservative with respect to type, charge, and polarity of residues. Predictions of secondary structure suggest that the size and position of alpha-helices are also conserved, even through pairwise amino acid identities between Haliotis rufescens and the different tegulines are less than 15%. PMID- 10368962 TI - Phylogenetic relationship of muscle tissues deduced from superimposition of gene trees. AB - Muscle tissues can be divided into six classes; smooth, fast skeletal, slow skeletal and cardiac muscle tissues for vertebrates, and striated and smooth muscle tissues for invertebrates. We reconstructed phylogenetic trees of six protein genes that are expressed in muscle tissues and, using a newly developed program, inferred the phylogeny of muscle tissues by superimposition of five of those gene trees. The proteins used are troponin C, myosin essential light chain, myosin regulatory light chain, myosin heavy chain, actin, and muscle regulatory factor (MRF) families. Our results suggest that the emergence of skeletal-cardiac muscle type tissues preceded the vertebrate/arthropod divergence (ca. 700 MYA), while vertebrate smooth muscle seemed to evolve independent of other muscles. In addition, skeletal muscle is not monophyletic, but cardiac and slow skeletal muscles make a cluster. Furthermore, arthropod striated muscle, urochordate smooth muscle, and vertebrate muscles except for smooth muscle share a common ancestor. On the other hand, arthropod nonmuscle and vertebrate smooth muscle and nonmuscle share a common ancestor. PMID- 10368963 TI - Performance of likelihood ratio tests of evolutionary hypotheses under inadequate substitution models. AB - In recent years, likelihood ratio tests (LRTs) based on DNA and protein sequence data have been proposed for testing various evolutionary hypotheses. Because conducting an LRT requires an evolutionary model of nucleotide or amino acid substitution, which is almost always unknown, it becomes important to investigate the robustness of LRTs to violations of assumptions of these evolutionary models. Computer simulation was used to examine performance of LRTs of the molecular clock, transition/transversion bias, and among-site rate variation under different substitution models. The results showed that when correct models are used, LRTs perform quite well even when the DNA sequences are as short as 300 nt. However, LRTs were found to be biased under incorrect models. The extent of bias varies considerably, depending on the hypotheses tested, the substitution models assumed, and the lengths of the sequences used, among other things. A preliminary simulation study also suggests that LRTs based on parametric bootstrapping may be more sensitive to substitution models than are standard LRTs. When an assumed substitution model is grossly wrong and a more realistic model is available, LRTs can often reject the wrong model; thus, the performance of LRTs may be improved by using a more appropriate model. On the other hand, many factors of molecular evolution have not been considered in any substitution models so far built, and the possibility of an influence of this negligence on LRTs is often overlooked. The dependence of LRTs on substitution models calls for caution in interpreting test results and highlights the importance of clarifying the substitution patterns of genes and proteins and building more realistic models. PMID- 10368964 TI - Diagnostic amino acids in actin genes: an idea whose time has gone. PMID- 10368965 TI - Incipient GAA repeats in the primate Friedreich ataxia homologous genes. PMID- 10368967 TI - Genetic variation of Cryphonectria hypoviruses (CHV1) in Europe, assessed using restriction fragment length polymorphism (RFLP) markers. AB - A total of 72 hypovirus-infected isolates of the chestnut blight fungus Cryphonectria parasitica were sampled from nine European countries between 1975 and 1997. The double-stranded RNA of the Cryphonectria hypoviruses (CHV1) was isolated and reverse transcription (RT)-PCR products were obtained for two different regions of the viral genome (ORF A and ORF B) using primer sequences of the type species CHV1-EP713. Both PCR products of each viral isolate were digested with four restriction endonucleases recognizing sequences of four nucleotides. The restriction fragment length polymorphism (RFLP) analysis revealed 41 genetically distinct RFLP types of CHV1 with 10 types occurring more than once. Identical RFLP types were detected nine times among viruses collected in the same location. Cluster analysis based on the RFLP banding patterns separated the viral isolates into five CHV1 clusters or subtypes. Most viral isolates (64 out of 72) grouped into one large cluster which comprised all viruses from Italy (including CHV1-EP747), Switzerland, Crotia, Bosnia, Hungary, Greece, and the French island Corsica, as well as five out of 11 isolates from continental France. Two additional subtypes of CHV1 were found in France (one related to CHV1-EP713) and one each in Spain and Germany. The Swiss samples collected over a period of 20 years showed that very little RFLP variation has evolved during this time. The results of this study are consistent with the hypothesis of multiple introductions of CHV1 into Europe. PMID- 10368969 TI - Reliable noninvasive genotyping based on excremental PCR of nuclear DNA purified with a magnetic bead protocol. AB - A new protocol for extraction of DNA from faeces is presented. The protocol involves gentle washing of the surface of the faeces followed by a very simple DNA extraction utilizing the wash supernatant as the source of DNA. Unlike most other protocols, it does not involve the use of proteinase K and/or organic extraction, but is instead based on adsorption of the DNA to magnetic beads. The protocol was tested by microsatellite genotyping across six loci for sheep and reindeer faeces. Comparison with DNA extracted from blood demonstrated that the protocol was very reliable, even when used on material stored for a long time. The protocol was compared with another simple, solid-phase DNA-binding protocol, with the result that the bead-based protocol gave a slightly better amplification success and a lower frequency of allelic drop-outs. Furthermore, our experiments showed that the surface wash prior to DNA extraction is a crucial step, not only for our protocol, but for other solid-phase protocols as well. PMID- 10368970 TI - Isolation of microsatellite markers in Astatoreochromis alluaudi and their cross species amplifications in other African cichlids. PMID- 10368971 TI - Characterization of six polymorphic microsatellite markers in gilthead seabream, Sparus aurata (Linnaeus 1758). PMID- 10368972 TI - Microsatellite markers for the onychophoran Euperipatoides rowelli. PMID- 10368973 TI - Characterization of polymorphic brown lemur (Eulemur fulvus) microsatellite loci and their amplification in the family Lemuridae. PMID- 10368974 TI - Characterization of hypervariable microsatellites in the cooperatively breeding white-browed sparrow weaver Plocepasser mahali. PMID- 10368976 TI - Quantitation of the acid and lactone forms of atorvastatin and its biotransformation products in human serum by high-performance liquid chromatography with electrospray tandem mass spectrometry. AB - A method for simultaneous quantitation of both the acid and lactone forms of atorvastatin, a new synthetic inhibitor of HMG-CoA reductase that is being marketed for the treatment of high serum cholesterol, and both the acid and lactone forms of its two biotransformation products, 2-hydroxyatorvastatin and 4 hydroxyatorvastatin, in human serum (a total of six analytes) by high-performance liquid chromatography with electrospray tandem mass spectrometry was developed and validated. A deuterium labeled analog was used as internal standard for each of the six analytes. Each point of the calibration standard curve, which ranged from 0.5 to 200 ng/mL, contained the six analytes at equal concentrations. Three groups of quality control (QC) samples were used. In the first group, combination QC samples contained all six analytes at equal concentrations. In the second group, acid-only QC samples contained only the acid forms (i.e. three analytes) at equal concentrations. In the third group, lactone-only QC samples contained only the lactone forms (i.e. three analytes) at equal concentrations. After adding the internal standard to 0.5 mL of each standard and the QC sample kept at 4 degrees C, the samples were acidified with sodium acetate buffer (pH 5.0) and then extracted with methyl tert-butyl ether. Detection was by positive ion electrospray tandem mass spectrometry using eight selected reaction monitoring channels. The acid compounds were stable in human serum at room temperature but the lactone compounds were unstable as they hydrolyzed rapidly to their respective acid forms. The conversion of the lactone compounds in both QC and post-dose human serum samples was nearly complete after 24 h at room temperature. The lactone compounds in serum could be stabilized by lowering the working temperature to 4 degrees C or lowering the serum pH to 6.0. The acid-only and the lactone-only QC samples showed that, under the sample processing conditions used, the degree of the hydrolysis of the lactone compounds or the lactonization of the acid compounds during the assay procedure was minimal (< 5%). The intra-day C.V., inter-day C.V. and the deviations from the nominal concentrations for all six analytes were within 15%, demonstrating good precision and accuracy. The required lower limit of quantitation (LLQ) of 0.5 ng/mL was achieved for each analyte. PMID- 10368977 TI - Characterization of tyrosine sulfate residues in antihemophilic recombinant factor VIII by liquid chromatography electrospray ionization tandem mass spectrometry and amino acid analysis. AB - Recombinant Factor VIII (rFVIII) is involved in the cascade of biochemical reactions leading to blood coagulation and is used for the treatment of haemophilia A. Plasma-derived FVIII (pdFVIII) has been reported to be post translationally modified by sulfation of tyrosine residues at positions 346, 1664, 1680, 718, 719 and 723. This report describes the quantitation of tyrosine sulfate residues in BHK-derived, human rFVIII by amino acid composition analysis and the identification of their positions in the polypeptide sequence using a combination of liquid chromatography and electrospray ionization mass spectrometry in the analysis of proteolytic digests of the protein. PMID- 10368978 TI - A new liquid chromatography/tandem mass spectrometric approach for the identification of class I major histocompatibility complex associated peptides that eliminates the need for bioassays. AB - Cell-surface class I major histocompatibility complex (MHC) molecules present processed self- and nonself-peptides to thymus-derived (T) lymphocytes, allowing the intracellular compartment of cells to be sampled in order to detect infection. Since the class I MHC-peptide complex plays a critical role in cell mediated immunity, it is important to obtain sequence information on the MHC associated peptides unique to infected cells as a prelude to the development of vaccines. Here, we outline and test an alternative strategy for identifying the proteins that are processed through the MHC pathway. This new strategy eliminates the necessity of developing and maintaining cytotoxic T lymphocyte (CTL) lines for peptide identification. In this new approach genome sequences from the infecting agent are scanned for stretches of amino acids that match a particular MHC binding motif. Molecular masses from these putative MHC-binding peptide sequences are calculated and compared to those found for peptides isolated from pathogen-infected host cells using liquid chromatography/mass spectrometry (LC/MS). Peptides with masses matching those in the database are then analyzed by tandem mass spectrometry (MS/MS) to determine their identity. Using this approach we were able to confirm the processing and presentation of two Trypanosoma cruzi proteins by the MHC class I pathway. These data suggest that a rigorous approach employing two-dimensional separations in conjunction with MS/MS and bioinformatics is a feasible means of identifying pathogen gene products of immunological interest when a CTL assay is unavailable or unsuccessful. PMID- 10368979 TI - Differentiation of 'isobaric' peptides and human milk oligosaccharides by exact mass measurements using electrospray ionization orthogonal time-of-flight analysis. AB - Exact mass determination is performed by electrospray ionization orthogonal time of-flight mass analysis. For peptides in the mass range of 1200-1500 Da a mass error of < 5 ppm is achieved with internal calibration within a single mass measurement provided peak intensities are high. Peptides containing isobaric amino acids like glutamine and lysine can thus be easily differentiated by their mass. In cases where more than one of these isobaric amino acids are present, the position of the amino acid can be revealed by exactly determining the mass differences between adjacent Yi" fragment ions in the collision-induced dissociation spectrum. Mass determination accuracy can be enhanced to 0.5-2 ppm by averaging over 8-10 mass measurements. Thereby compositional analysis of human milk oligosaccharides in a mixture can be performed in the mass range up to 3000 Da, even for low-intensity molecule-ion signals and for isobaric compounds with a mass difference of only 0.025 Da. PMID- 10368980 TI - Fingerprinting of Helicobacter pylori strains by matrix-assisted laser desorption/ionization mass spectrometric analysis. AB - Helicobacter pylori is an important human gastrointestinal pathogenic bacterium which is believed to colonize approximately one-half of the world's population. Different strains of H. pylori possess virulence proteins for tissue colonization, host evasion and tissue damage. The bacteria display genomic instabilities that include gene rearrangements and gene exchange. Recently, methods for matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) have been established for monitoring biomarkers in bacterial extracts. In order to establish a set of H. pylori specific biomarkers as well as a set of strain-specific biomarkers, we examined lysates and extracts from six different strains of this bacterium. Three different MALDI matrices, alpha-cyano-4-hydroxycinnamic acid, sinapinic acid, and ferulic acid were tested for sensitivity of analysis. Also, the effects of solubilizing analytes with the detergent n-octyl-beta-D-glucopyranoside were explored. It was found that a set of H. pylori specific, and a probable set of strain-specific, biomarkers could be established using MALDI-TOFMS. The use of H. pylori fingerprinting by MALDI may be useful for typing of these bacteria, or for studying genetic drift at the phenotypic level in specific strains. PMID- 10368981 TI - Escherichia coli lipopolysaccharide effects on proliferating rat liver cells in culture: a morphological and functional study. AB - Liver cells have been implicated in playing an important role in the pathogenesis of endotoxic shock-associated liver injury. The present study was designed to investigate Escherichia coli 0111:B4 lipopolysaccharide (LPS) effects on proliferating rat liver cells in culture. Isolated cell system can rarely serve as models of complete organisms, but with using an in vitro test-model, endogenic factors (e.g. hormonal effects, nervous influences, blood activation, etc.) or experimental stress in laboratory animals can be eliminated as variables affecting hepatocellular responses. In the proposed in vitro model, using specially proliferating liver cells, morphological cell alterations were observed after 30 min and low doses endotoxin administration (10 micrograms/ml). LPS induced an extreme fragility and a diminished adhesion capacity on cells. Cell-to cell contacts were also disturbed. The LPS treatment produced extreme heterogeneity in liver cell size, enlargement of nuclei, nuclear and cytoplasmic protrusions, and increased the number of large nucleoli and lipid droplets, also decreasing the angiotensin action on intracellular calcium levels. The effects observed after the LPS exposition can be related with an altered metabolism of the liver cells in culture due to a destabilization of plasma membrane, a transmembrane signalling alteration, and a mitochondrial damage. The specificity of cell response to endotoxic lipopolysaccharide suggests a multiple membrane damage inducing important metabolic disturbances. PMID- 10368982 TI - Age-related changes and location of type I, III, IV, V and VI collagens during development of four foetal skeletal muscles of double-muscled and normal bovine animals. AB - The aim of this work was to study the variability in the quantities of hydroxyproline, of type I and III collagens and in the location of types, I, III, IV, V, VI in four muscles of normal and double-muscled (DM) cattle. Samples were collected from foetuses at different ages post-conception. Both in the two genetic types and in muscles, from 110 days, types I, III, V, VI were located in perymisium and types I, IV, V, VI in endomysium. The amounts of hydroxyproline and of type I collagen increased from 150 to 180 or 230 days then decreased up to 260 days, with a trend to lower quantities in muscles of DM animals. Depending on the muscle and of the genetic type, amounts of type III, or changed as those of type I, or remained stable. Whatever the genetic type, at the end of gestation, proportions of type I and III in the total collagen are not identical in the four muscles, differences between muscles being particularly marked for type III, CT and MA muscles being the richer in this type. In addition, these two muscles contained less type III in DM animals than in normal ones. PMID- 10368983 TI - Follistatin possesses trunk and tail organizer activity and lacks head organizer activity. AB - In this report the organizer activity of follistatin was examined by transplantation of pieces of the animal cap, isolated from embryos injected with follistatin mRNA, into the blastocoele of an early host blastula (Einsteck explants). Host embryos developed a secondary axis consisting of myotomes, notochord and neural tube of the trunk or tail character. Secondary structures that are characteristic of a head, such as cement glands or brain and eyes, did not develop in these experiments. These findings suggested that follistatin may have the trunk and tail organizer activity, while it was not possible to reconstitute its head organizer activity. PMID- 10368984 TI - Lymphatic endothelium of the human tongue expresses multiple leukocyte adhesion molecules. AB - The expression of leukocyte adhesion molecules on lymphatic vessels of the human tongue was examined using histochemical and immunohistochemical methods. Three different types of lymphatic vessels were distinguished: type I vessels expressed intercellular adhesion molecule-1 (ICAM-1), platelet-endothelial cell adhesion molecule-1 (PECAM-1), and endothelial cell-selectin (ELAM-1); type II vessels expressed ICAM-1 and PECAM-1; and type III vessels expressed PECAM-1 only. The lymphatic vessels located very close to the oral epithelium (lymphatic capillaries) and the other lymphatic vessels near the oral epithelium were type I. The lymphatic vessels in the submucosal connective tissue (collecting lymphatic vessels) were type II and type III. The results suggest that there may be functional differences in the lymphatic endothelium, where lymphatic capillaries are more active than collecting lymphatic vessels in lymphocyte migration from tissue into the lymphatic vessels. PMID- 10368985 TI - Functional distribution and further characterization of human endothelial ligand for cellular L-selectin. AB - In the present study we examine the functional distribution of the human endothelial L-selectin ligand, which determines the sites of extravasation of L selectin-positive cells. A murine cell line transfected with human L-selectin adhered preferentially to the high endothelial venules (HEV) of human peripheral lymph nodes compared to the HEV of mucosal lymphoid tissues (mean of 0.83 compared to a mean of 0.07 cells per HEV respectively). In addition, an antibody against L-selectin differentially inhibited the adhesion of human lymphocytes to peripheral lymphoid tissue versus mucosal lymphoid tissue HEV (mean 41 and 5% inhibition respectively). Although both sulfoglucuronyl-containing glycolipids and sialyl-Lewis X have been proposed as endothelial ligands for L-selectin, an antibody against the former did not bind to peripheral lymph node endothelium, and an antibody against the latter did not block adhesion of L-selectin expressing cells. The enzyme O-sialoglycoprotein endopeptidase caused up to an 84% reduction in L-selectin-dependent binding, indicating that sialylated glycoproteins containing O-linked glycans are essential for a large majority of adhesion via L-selectin. PMID- 10368986 TI - Immunohistochemical localization of epididymal secretory glycoprotein EP1 in the adult male chimpanzee. AB - Proteins, synthesized by the epididymal epithelium, are secreted sequentially into the lumen of the ducts epididymis where they effect sperm maturation and enable functional motility and fertilizing capacity. EP1 is a major secretory glycoprotein of chimpanzee (Pan troglodytes) epididymis. The epididymal duct exhibits diverse histology (Smithwick & Young, 1997). Epithelia I-V of the efferent ducts show no characteristic anti-EP1 binding. The densest granules of anti-EP1 reaction product appear in epithelium VI adjacent to the basal lamina in the infranuclear region of the principal cells (PCs), in the cytoplasm of the apical half of the PCs, and in the perinuclear and perivacuolar cytoplasm of the basal cells. In epithelia VII-XIV of the ductus epididymis proper, anti-EP1 binding decreases distally and is localized in the cytoplasm of the PCs and basal cells, among the stereocilia of the luminal border, within various microvillar borders, and in the luminal fluid. Therefore, EP1 appears to be synthesized and secreted primarily in the caput region of the ductus epididymis and may be reabsorbed nonselectively across epithelia with apical microvilli, including the non-ciliated cells of efferent ducts, the distal corpus and cauda of the ductus epididymis, and the proximal ductus deferens. PMID- 10368987 TI - Effect of subcutaneous pancreatic tissue transplants on streptozotocin-induced diabetes in rats. I. Morphological studies on normal, diabetic and transplanted pancreatic tissues. AB - The present study examines the morphological changes occurring in subcutaneous pancreatic tissue grafts (SPTG) and its effect on the host pancreatic islet cells in streptozotocin (STZ)-induced diabetic rats using morphological techniques. SPTG survived after 15 weeks of transplantation. Its acinar cells degenerated but the ducts and endocrine cells survived. The surviving and newly formed pancreatic tubules and endocrine cells filled the spaces left by degenerated acinar cells. Compartmentalization of the surviving parenchymatic tissues was observed, with the pancreatic tubules lying in the periphery of the graft and the endocrine tissue in the inner portion of the graft. Lymphocytes invaded the inner portion of the graft, conglomerating around endocrine cells. It was interesting, however, that, lymphocytes where not observed in the periphery of the grafts where most of the surviving pancreatic tubules lie. In addition to this, necrotic tissues were observed in the inner part of the graft. Fifteen weeks after transplantation into the subcutaneous region, insulin, glucagon, somatostatin and pancreatic polypeptide-immunoreactive cells were observed in many parts of the graft. In the peripheral parts of the grafts, large numbers of pancreatic tubules differentiated into endocrine cells. In conclusion, the ductal and endocrine cells of pancreatic tissue fragments survived in the subcutaneous region of rat with normal pattern of distribution. PMID- 10368988 TI - Effect of subcutaneous pancreatic tissue transplants on streptozotocin-induced diabetes in rats. II. Endocrine and metabolic functions. AB - The present study examines the effect of subcutaneous pancreatic tissue grafts (SPTG) on endocrine and metabolic functions in streptozotocin (STZ)-induced diabetic rats using radioimmunoassay and biochemical techniques. SPTG survived even after 15 weeks of transplantation and significantly improved the weight of STZ-diabetic rats over a 15-week period. Although blood glucose-, cholesterol-, and glycosylated-haemoglobin (GHb) levels were not significantly lower in STZ diabetic rats treated with SPTG, the values of these biochemical parameters were lower than those in untreated diabetic rats. Plasma and pancreatic immunoreactive C-peptide (IRCP) levels did not improve after SPTG (IRCP expressed as mean +/- standard deviation were 0.22 +/- 0.07, 0.072 +/- 0.02 and 0.08 +/- 0.03 pg ml-1 in the plasma non-diabetic diabetic and treated rats respectively, while IRCP levels in the pancreas of the non-diabetic, diabetic and treated rats were 433.8 +/- 0.1, 22.9 +/- 0.01 and 10.4 +/- 0.01 pg mg tissue-1 respectively). SPTG, however, improved plasma immunoreactive insulin (IRI) levels in both plasma and pancreas. IRI values in plasma were 54.7 +/- 13.6, 18.0 +/- 5.0 and 22.1 +/- 4.3 microUI ml-1 in non-diabetic, diabetic and treated rats respectively and were 277.3 +/- 37.1, 14.7 +/- 1.8 and 30.3 +/- 15.9 microIU micrograms tissue-1 in the pancreas of non-diabetic, diabetic and treated rats respectively. There was improvement in immunoreactive glucagon (IRG) levels after SPTG. IRG values in the plasma of non-diabetic, diabetic and treated rats were 147.0 +/- 10.7, 408.0 +/- 76.5 and 247.7 +/- 3 pg ml-1 respectively whereas, IRG measured in the pancreas was 1642.25 +/- 424.23, 1899.0 +/- 290.4 and 1714.1 +/- 301.98 pg micrograms tissue-1 in non-diabetic, diabetic and treated rats, respectively. The pancreas:plasma ratio of pancreatic hormones was deranged in untreated diabetes but improved after SPTG. In conclusion, SPTG significantly improved the weight gain, pancreatic insulin content, plasma IRG and pancreas: plasma ratio of IRCP, IRI and IRG. It also reduced blood glucose-, cholesterol-, and glycosylated hemoglobin levels in STZ-diabetic rats. PMID- 10368989 TI - Effect of subcutaneous pancreatic tissue transplants on streptozotocin-induced diabetes in rats. III. Distribution of neuropeptides in normal and diabetic (host) pancreas. AB - This study examines whether there is a change in the pattern of distribution of cholecystokinin-octapeptide (CCK-8), calcitonin-gene-related peptide (CGRP), neuropeptide-Y (NPY), substance P (SP) and vasoactive intestinal polypeptide (VIP) in the pancreas of streptozotocin (STZ)-diabetic (host) rats after subcutaneous pancreatic transplantation. Varicose CCK-8-immunopositive nerve fibres were observed in the wall of blood vessels of both normal and diabetic host pancreata. The density of CCK-8-immunoreactive varicose nerve fibres appeared to have increased in host rat pancreas. CGRP was demonstrated in many nerve fibres located in the wall of blood vessels of both normal and host pancreas. CGRP, however, seemed to be better expressed in the nerves of host pancreas when compared to normal. The pancreata of both normal and diabetic (host) rats contained numerous NPY-immunopositive varicose nerve fibres located in the wall of blood vessels. SP was demonstrated in neurons located in the interlobular areas of normal tissue and in fine varicose nerve fibres of the interacinar region of the pancreas of STZ-induced diabetic rats with SPTG. In normal pancreatic tissue, VIP-immunopositive nerve fibres were observed in all areas of the pancreas. VIP-positive nerve fibres were still discernible especially in the interacinar regions of the pancreas of host rats. In conclusion, the pattern of distribution and density of NPY, SP and VIP in the pancreas of STZ-induced diabetic rats with SPTG is similar to that observed in normal pancreas, but the expression of CGRP and CCK-8 seemed to have increased as a result of transplantation and or diabetes. PMID- 10368990 TI - Ultrastructure of peritubular tissue in association with tubular hyalinization in human testis. AB - Testicular peritubular tissue, also known as the tunica propria, surrounds the seminiferous tubules and is responsible for contractile, paracrine and transport functions. The aim of the present report is to describe the pathomorphology of peritubular tissue in association with tubular hyalinization in human testis. Twenty-seven testicular biopsies from 21 subfertile and infertile men were studied with the electron microscope. Biopsies from five patients showed complete or nearly complete tubular hyalinization. In addition to changes described earlier, the following new ultrastructural features were observed: 1. loss of polarity and configuration of myoid cells; 2. protrusion of myoid cells towards the tubule and evagination of basal lamina surrounding the tubule towards the interstitial direction leading to 'bridge' formation. These 'bridges' of myoid cells often created completely separated small compartments within the tunica propria; 3. vacuolization and fragmentation of myoid cell nuclei; 4. a balloon like swelling of myoid cell containing phagolysosomes and lipid droplets. We conclude that disorganization and loss of vital functions of the extracellular matrix and myoid cells contribute to the pathogenesis of tubular hyalinization. PMID- 10368992 TI - [The development of microwave coagulation therapy for hepatocellular carcinoma]. PMID- 10368991 TI - Light microscopic observations on the reproductive tract of the male sand rat, Psammomys obesus. AB - The anatomy of the reproductive tract of the male sand rat, Psammomys obesus, was examined by light microscopy. Histologically, the reproductive tract is similar to other rodent species. Seminiferous tubules in the 1-month-old sand rat do not contain a tubular lumen but Sertoli cells, spermatogonia and spermatocytes are present. A full complement of germ cells is present in the seminiferous tubules by 2.5 months and spermatogenesis is well established. The interstitial space is not well defined until 2.5 months when cell types typical of most rodent species are observed. The epididymis is not noticeably segmented into lobules. An epididymal lumen is not observed until 2.5 months. Cauda epididymal sperm are not observed in the 1 or 2.5-month-old animals and cauda epididymal sperm counts from the 7.5 and 12.5-month-old animals are highly variable. The epididymis, proximal and middle regions of the vas deferens, seminal vesicles and prostate display morphological and histological characteristics similar to other rodent species. The distal end of the vas deferens is not expanded to form an ampulla. PMID- 10368993 TI - [Comparison of the therapeutic effects and acid suppression of H2-receptor antagonist and proton pump inhibitor in patients with gastric body ulcer--a prospective controlled trial]. AB - We studied, the rate and the affecting factor of ulcer healing of patients with gastric body ulcer treated by H2-receptor antagonist (H2-RA) and proton pump inhibitor (PPI). Gastric acidity was also examined using 24 hr pH monitoring. No difference was observed between the affecting factor of ulcer healing and healing rate (94.7% and 94.9%) among the patients treated by H2-RA or PPI. The average time below pH3 during treatment with H2-RA or PPI were 17.4 +/- 4.3 hr and 23.0 +/- 1.5 hr, respectively. Acid suppression was superior in PPI treated group than in H2-RA group. From these findings, we concluded that H2-RA had sufficient therapeutic efficacy in treating gastric body ulcer. PMID- 10368994 TI - [The clinicopathological analysis of lymph node metastasis of gastric cancer]. AB - One hundred-twenty-one cases of curative resection for gastric cancer with lymph node metastasis were analyzed to determine the prognostic value of the nodal stage (n), number of metastatic lymph nodes, maximum diameter of metastatic lymph nodes, micrometastasis of lymph nodes, histological type of lymph node metastasis, extranodular invasion and infiltration into lymphatic vessels around metastatic lymph nodes. In patients with a higher nodal stage the survival rate was lower and the nodal stage was a good prognostic indicator. Inpatients who had 7 or more metastatic lymph nodes or in whom the maximum diameter of the metastatic lymph nodes was over 15 mm or who had not only micrometastasis of lymph nodes, the survival rate was lower. These results suggest that quantitative analysis of metastatic lymph nodes is necessary. In patients who had nudifferentiated type metastatic lesions of lymph nodes, the survival rate was lower than in those with the differentiated type and the difference was larger than for the primary lesion. In patients who had extranodular invasion or infiltration into lymphatic vessels around metastatic lymph nodes, the survival rate was decreased. These results suggest that histopathological analysis of metastatic lymph nodes is necessary. PMID- 10368995 TI - [Endoscopic and pathological changes of gastric lesion before onset, during and after treatment in a patient with Cronkhite-Canada syndrome]. PMID- 10368996 TI - [Acute myocarditis due to mesalazine in a patient with ulcerative colitis]. PMID- 10368997 TI - [A case of simple liver cyst with markedly elevated CEA level in the cystic fluid]. PMID- 10368998 TI - [Occurrence of hepatocellular carcinoma in patients with chronic hepatitis C who continued to be HCV-RNA negative for up to 3 years by interferon]. PMID- 10368999 TI - [A case of drug-induced fulminant hepatic failure accompanied with marked increase of pancreatic enzymes]. PMID- 10369000 TI - [A case of primary biliary cirrhosis associated with pernicious anemia]. PMID- 10369001 TI - [A case of lymphoepithelial cyst of the pancreas]. PMID- 10369002 TI - [A case of pancreatoblastoma in an adult]. PMID- 10369004 TI - Educational programs for deaf students. Schools and programs in the U.S. PMID- 10369003 TI - The functional assessment of deaf and hard of hearing students. AB - The study reports on a set of questions added to the 1997-98 Annual Survey of Deaf and Hard of Hearing Children and Youth designed to take into consideration the functioning of children in their classroom in nine functional areas. Basing information on 30,198 students, the study describes prevalence rates of reported limitations in these functional areas for deaf and hard of hearing students, compares these to rates resulting from the reporting of categorically defined additional disabilities, and examines interrelationships among the items. Results of school estimates of students' functional hearing abilities are presented. The study's findings suggest a broader range and higher prevalence of functional limitations than would be assumed by analyzing categories of additional disabilities alone. The study's findings support the viability of functional assessment through large surveys. The discussion emphasizes the importance of functional assessment for the provision of appropriate educational services to deaf and hard of hearing children. PMID- 10369005 TI - Educational programs for deaf students. Schools and programs in Canada. PMID- 10369007 TI - University and college programs for personnel in deafness. PMID- 10369006 TI - Educational programs for deaf students. Postsecondary programs. PMID- 10369008 TI - Programs for deaf-blind children and adults. PMID- 10369009 TI - Educational programs for deaf students. Supportive and rehabilitative programs. PMID- 10369010 TI - Research on deafness. Doctoral dissertations. PMID- 10369011 TI - Francesco Rizzoli's dream (1809-1880). PMID- 10369012 TI - Changes in the femoral cortex as related to prosthetic stem. AB - The need to improve the features of resistance and duration of prosthetic implants, based on the constant increase in the number per year of total hip substitutions has encouraged the study of the causes of aseptic loosening of implants, in particular, any relationship between femoral morphology, features of the implanted prosthesis (size and coating), degree of contact between the latter and the host bone, and the occurrence of changes in bone trophism. The documentation relative to 143 patients, corresponding to 149 primary hip arthroplasties, was evaluated clinically (according to Merle d'Aubigne modified by Charnley) and radiographically, using a computerized radiographic evaluation system devised by our group. In particular, on x-rays (anteroposterior view) obtained in the area corresponding to the upper and lower margins of the smaller trochanter and of the femoral isthmus, measurements relative to femoral diameters, to the distance between them, and to the corresponding flare indexes were obtained; bone thickness, degree of bone-prosthesis correspondence and any changes in bone trophism caused by stress-shielding and stress-concentration were also measured. An analysis of the data was carried out using non-parametric statistical tests, that allowed us to reveal the surgeon's good standardization in preparation of the femur; the tendency for the cortex to thicken for prostheses of the short type, and the narrowing of the standard type prostheses, the influence of the degree of fit of the prosthesis on trophism of the femoral cortex. PMID- 10369013 TI - Evaluation of homoplastic bone graft in hip replacement revision surgery. AB - A total of 17 patients submitted to hip replacement revision surgery in our clinic between 1986 and 1994 were evaluated in order to analyze the effectiveness of homoplastic support grafting. Loss of substance involved the acetabulum and the femur. Four parameters were evaluated radiographically: fusion time, restructuring time, flattening or collapse of grafts, and, finally, graft resorption. The results showed that the homoplastic graft is characterized by excellent biological behavior, and that the success of the new implant depends mostly on surgical method used as the graft was always integrated quickly and completely. In cases with considerable loss of substance it is important to use means of support that allow for partial unloading of the forces between prosthesis and host bone, thus avoiding excessive loading in the grafts. PMID- 10369014 TI - Functional evaluation in total hip replacement patients. AB - An evaluation of total hip replacement patients requires objective criteria that allow for the measurement of any changes in the biomechanics of the prosthesis and the effects of these changes on clinical findings. An evaluation of loading and joint movement during walking in these patients has been dealt with by many authors. In this study 15 patients who had undergone total hip replacement for coxarthrosis primary or secondary to congenital dysplasia were examined by kinematic and kinetic gait analysis, clinical evaluation, and radiographic evaluation of the position of the neocenter of prosthetic joint rotation. The patients with prostheses in congenital dysplasia presented with a pattern of walking that was significantly modified. In agreement with what is reported in the literature proximal positioning had less influence on gait parameters as compared to lateral dislocation. Furthermore, changes in positioning observed did not seem to have negative effects on joint loading, measured indirectly by the calculation of joint moments. PMID- 10369015 TI - Loss of bone mass after total hip replacement: preliminary data. AB - In a total of 37 females with cemented total hip replacement for monolateral coxarthrosis, of which 13 with prosthetic stem loosening, and 11 with monolateral coxarthrosis that is not prosthetized, bone mineral density (BMD) is determined by dual ray photonic absorbimetry selecting regions of interest (ROI) on the cortex of the femurs 4 cm under the lesser trochanter and on the ischium bilaterally. In females that are not prosthetized there are differences in bone mass between the two femurs and the ischium on both sides. In prosthetized patients BMD of the femur and of the ischium on the side operated on is significantly less than on the contralateral one (Student's "t" test: p < 0.001). In patients with stable prostheses, BMD of the femur operated on is greater than that in females with prosthetic stem loosening (Student's "t" test: p < 0.000). Based on a comparison between these two groups we did not observe any other significant differences in BMD among the ROI analyzed. BMD was correlated with the amount of time since surgery only in the ROIs in prosthetized femurs. The study confirms the significant bone resorption of the cortex in prosthetized femurs and documents analogously significant reduction in BMD in the ischium on the side operated on. Finally, it indicates that prosthetic stem loosening may be associated with loss of BMD in the femoral cortex which is significantly greater than that observed, during analogous periods of time in stable implants. PMID- 10369016 TI - The surgical treatment of spondylolisthesis with transpedicular stabilization: a review of 25 cases. AB - The present study concerns a series of 25 patients submitted to vertebral stabilization and fusion for the treatment of spondylolisthesis. In all of the cases a transpedicular screw system was used. The Steffee plates and screws system was used in 22 cases; the Cotrel-Dubousset (CD) system was used in 3. The listhetic vertebra was L5 in 14 cases, it was L4 in 11 cases. Based on Meyerding radiographic criteria there were: grade I: 8 cases, grade II: 13 cases, grade III: 4 cases. Instrumented fusion was performed at only 1 level in 20 cases, at 2 levels in 5 cases. Mean follow-up was 29 months. Clinical results were evaluated based on the White criteria. The results were satisfactory in 78% of cases. The present study shows that transpedicular vertebral stabilization constitutes a safe method, offering excellent stability essential to consolidation of fusion. PMID- 10369017 TI - Distal centering in locked intramedullary osteosynthesis of the femur: use of a magnet-resistant probe. AB - The authors present an original solution for distal screw fixation in locked nail intramedullary osteosynthesis, traditionally carried out with image intensifier. The system includes the determination of an electromagnetic field produced by a microbobbin placed in the hole and, thus, coaxial with it. The study, conducted through a probe equipped with special magnet-resistant sensors capable of picking up electromagnetic variations, is based on the same principle as a compass. In addition to being effective and simple, the method avoids exposure to ionizing radiations required for the search for holes. PMID- 10369018 TI - Osteosynthesis with percutaneous wiring in fractures of the proximal humerus. AB - The authors report the preliminary results of osteosynthesis with percutaneous wiring in the treatment of fractures of the proximal humerus. This method which is still described in very few cases, is characterized by several advantages such as its minor invasiveness, and respect for vascularization of the fragments that guarantee good functional results. The application of threaded pins or pins of large diameter guarantees excellent hold with a low incidence of loosening secondary to loss of reduction. The use of a traction device applied to the surgical table simplifies reduction maneuvers and osteosynthesis. PMID- 10369019 TI - The effects of metal corrosion debris on immune system cells. AB - Authors evaluated the correlation between immune system and metal ions release in blood of 17 subjects with Cr/Co/Ni joint prostheses. For the purpose Chromium (Cr), Cobalt (Co) and Nickel (Ni) serum levels were measured and, at the same time some immunological parameters (Leukocytes, Lymphocytes and Lymphocytes T, B and Natural Killer cells sub-populations) were evaluated. The results showed a significant decrease of Leukocytes, Lymphocytes and of T Lymphocytes sub populations. At the same time it was demonstrated a significant increase of Chromium, Cobalt and Nickel levels in patients with joint prostheses as compared to control population (23 patients). In conclusion, ions release from metallic surface of the prostheses is correlated with a depression of immune system. This correlation could depend on a toxic action on immune system caused by the products released by the implant. It could also depend on a lymphocytes compartimentalization in periprosthetic tissues as a consequence of a cell mediated hypersensitivity reaction towards implants corrosion products. PMID- 10369020 TI - Evaluation of cytokines regulating bone turnover in the serum of post-menopausal and senile women. AB - The authors evaluated the Insulin-like Growth Factor-1 (IGF-1) stimulating the formation of bone tissue, and interferon gamma (IFN gamma), inhibiting bone resorption, in the serum of women, 13 of fertile age, 34 of post-menopausal age, and 14 of senile age. Values for IGF-1 in the serum were considerably low in patients of post-menopausal and senile age, and presented highly significant differences with values for subjects of fertile age. The values for IFN gamma did not present significant differences between different age groups. It may be assumed that post-menopause and during senile age physiological osteopenia may be favored by a decrease in the secretion of IGF-1. PMID- 10369021 TI - Evolution of the concepts of functional anatomy of the knee joint. AB - It is the purpose of this study to present a review of studies on the kinematics of the knee joint and load distribution in the two compartments, medial and lateral, in order to reveal the evolution of concepts of functional anatomy. Flexion-extension features have been clearly defined by numerous authors who have used different methods of research. Transverse rotation, instead, still needs to be clarified. In particular, whether or not a screw-home movement exists has been questioned by many authors who have studied the kinematics of the knee, both without loading and during walking. Dynamic studies have shown how the radiographic method (static) to deduce load distribution in the knee joint is inadequate; this is because during walking, loading tends to be transmitted on the medial compartment, also the case in valgus knees. PMID- 10369022 TI - Acetabular osteoblastoma: description of a case. AB - Osteoblastoma is a slow-progressing, benign bone tumor, that is not frequently observed in clinical orthopaedics (approximately 1% of all primary bone tumors). There is predilection for the vertebrae (posterior arch), the femur, the tibia, and the cranium; it affects young subjects (from 10 to 35 years), with predilection for males (males: females = 2:1). Symptoms are not very specific, characterized essentially by moderate, discontinuous pain, that is responsive to treatment by NSAIDS; it may, at times, be asymptomatic. On radiographic assessment it is viewed as a lytic area that is rounded, greater than 2 cm in size, with unclear margins, with or without peripheral bone reaction. It is not easy to diagnose osteoblastoma, particularly if it is localized in unusual sites, such as in the pelvis. The authors present a case of osteoblastoma of the acetabular bottom in a subject aged 22 years, that was not diagnosed unrecognized for about 2 years from the onset of symptoms. PMID- 10369023 TI - The chondroepitrochlearis muscle: case report. AB - An extremely rare muscle disorder in the pectoral region is described. It involves a musculotendinous unit which originates from the pectoralis major and inserts onto the medial epicondyle of the humerus. The authors report a case of an 11-year-old boy, affected bilaterally, who underwent surgical treatment to lengthen the chondroepitrochlearis muscle. There was significant cosmetic and functional improvement. PMID- 10369024 TI - Recurrent dislocation of the elbow in children: description of a case. AB - The authors report a case of recurrent dislocation of the elbow in a boy aged 11 years, surgically treated by reconstruction of the lateral capsuloligamentous structures. After a review of the literature, the need for a diagnostic protocol to establish the main cause of recurrent dislocation is emphasized. PMID- 10369025 TI - Quiz. Bilateral intraosseous mucous cyst of the tibia. PMID- 10369026 TI - Interruption of the radial nerve at proximal level: reconstruction following anterior transposition. AB - The authors describe two cases of radial paralysis due to interruption in the proximal level, treated by autologous nerve grafting, following anterior transposition. At long-term follow-up, in one case after 12 years, there was good functional recovery, while in the second case there was still no recovery after 2 years. The surgical technique involving anterior transposition of the nerve is described, which in this type of lesion facilitates neurorrhaphy. PMID- 10369027 TI - The health and illness of the artists: Giacomo Leopardi's scoliosis. PMID- 10369028 TI - In memoriam. Mario Campanacci. PMID- 10369029 TI - Sex bias in the diagnosis of personality disorders: an evaluation of the DSM-IV criteria. AB - This study considered whether the Diagnostic and Statistical Manual of Mental Disorders (4th ed.; American Psychiatric Association, 1994) is biased against women by requiring less dysfunction for the personality disorders that are more commonly diagnosed in women (e.g., histrionic). Clinicians estimated the extent of social dysfunction, occupational dysfunction, and personal distress suggested by each of the diagnostic criteria for 6 personality disorders. The results failed to suggest a bias against women, as there was no difference in the overall level of dysfunction associated with the female-typed personality disorder diagnostic criteria (fewer criteria are also required for the male-typed diagnoses). However, the considerable variation in dysfunction across disorders and criteria, and the minimal degree of impairment implied by some of the diagnostic criteria, also raise more general issues that should perhaps be addressed in future editions of the diagnostic manual. PMID- 10369030 TI - Cognitive reactivity and depressotypic information processing in children of depressed mothers. AB - Although high-risk research suggests that children of depressed mothers are at increased risk for psychological disorders, the mechanisms of this risk are not well understood. In the current study, the information processing of children of depressed mothers was compared with that of children whose mothers were not depressed. Half of each group received a priming induction designed to activate cognitive schemas prior to assessment. All children then completed a self referent processing task that examined the recall of negative and positive information. Results indicate that when primed, at-risk children showed a less positive self-concept and more negative information processing than did the children in the other groups. These data may offer potential clues into the mechanisms of cognitive vulnerability in at-risk children. PMID- 10369031 TI - Social skills, social outcomes, and cognitive features of childhood social phobia. AB - Social skills, social outcomes, self-talk, outcome expectancies, and self evaluation of performance during social-evaluative tasks were examined with 27 clinically diagnosed social phobic children ages 7-14 and a matched nonclinical group. Results showed that, compared with their nonanxious peers, social phobic children demonstrated lower expected performance and a higher level of negative self-talk on social-evaluative tasks. In addition, social phobic children showed social skills deficits as assessed by self- and parent report, an assertiveness questionnaire, and direct behavioral observation. Furthermore, compared with the control group, social phobic children were rated by themselves and others as significantly less socially competent with peers and were found to be less likely to receive positive outcomes from peers during behavioral observation. Implications for the assessment and treatment of childhood social phobia are discussed. PMID- 10369032 TI - Preattentive bias for emotional information in panic disorder with agoraphobia. AB - A combined emotional Stroop and implicit memory (tachistoscopic identification) task with 3 types of words (panic-related, interpersonal threat, and neutral words) and 2 exposure conditions (subliminal, supraliminal) was administered to 35 patients with panic disorder and 35 age- and sex-matched controls. The patients showed Stroop interference for panic-related words both sub- and supraliminally and a similar but not equally robust effect on interpersonal threat words. On the tachistoscopic identification task, the patients identified more panic-related words than the controls did but showed no implicit memory bias effect. The patients' subliminal Stroop interference for panic-related words was found to correlate with trait anxiety and depression, although not with anxiety sensitivity. PMID- 10369033 TI - Perceptual asymmetry differences between major depression with or without a comorbid anxiety disorder: a dichotic listening study. AB - Predictions that anxious and nonanxious depression would differ in perceptual asymmetry (PA), as well as in sensitivity for perceiving emotional words, were evaluated using dichotic listening tasks. A total of 149 patients having a major depressive disorder (51 with and 98 without an anxiety disorder) and 57 healthy controls were tested on fused-word and complex tone tasks. The anxious and nonanxious depression groups showed a consistent difference in PA across tasks; that is, the anxious group had a larger left-ear advantage for tones and a smaller right-ear advantage for words when compared with the nonanxious group. There was no group difference in sensitivity for perceiving emotional words. Patients having an anxious depression appear to have a greater propensity to activate right than left-hemisphere regions during auditory tasks, whereas those having a nonanxious depression have the opposite hemispheric asymmetry. PMID- 10369034 TI - Manipulating smoking motivation: impact on an electrophysiological index of approach motivation. AB - A chief goal of this research was to determine whether stimuli and events known to enhance smoking motivation also influence a physiological variable with the potential to index approach motivation. Asymmetry of electroencephalographic (EEG) activity across the frontal regions of the 2 hemispheres (left minus right hemisphere activation) was used to index approach motivation. In theory, if EEG asymmetry sensitively indexes approach dispositions, it should be influenced by manipulations known to affect smoking motivation, that is, exposure to smoking cues and tobacco deprivation. Seventy-two smokers participated in this research and were selectively exposed to a smoking-anticipation condition (cigarettes plus expectation of imminent smoking) following either 24 hr of tobacco withdrawal or ad libitum smoking. Results indicated that EEG asymmetry was increased by smoking anticipation and that smoking itself reduced EEG asymmetry. Results also suggested that smoking anticipation increased overall (bihemispheric) EEG activation. Results were interpreted in terms of major theories of drug motivation. PMID- 10369035 TI - Friendship clique and peer influences on body image concerns, dietary restraint, extreme weight-loss behaviors, and binge eating in adolescent girls. AB - This study explored friendship variables in relation to body image, dietary restraint, extreme weight-loss behaviors (EWEBs), and binge eating in adolescent girls. From 523 girls, 79 friendship cliques were identified using social network analysis. Participants completed questionnaires that assessed body image concerns, eating, friendship relations, and psychological family, and media variables. Similarity was greater for within than for between friendship cliques for body image concerns, dietary restraint, and EWLBs, but not for binge eating. Cliques high in body image concerns and dieting manifested these concerns in ways consistent with a high weight/shape-preoccupied subculture. Friendship attitudes contributed significantly to the prediction of individual body image concern and eating behaviors. Use of EWLBs by friends predicted an individual's own level of use. PMID- 10369036 TI - The sexual preferences of incest offenders. AB - Inclusive fitness theory suggests that discriminative solicitude and inbreeding avoidance are important mechanisms regulating parent-children interactions. From an inclusive fitness perspective, sex with one's biological children is paradoxical. The authors hypothesized that incest can occur when these mechanisms are not activated (e.g., if a father is uninvolved in child rearing) or are overwhelmed by another factor, such as pedophilic interest. They predicted that biological fathers, who presumably have been the most involved in the rearing of their victims, would show greater phallometrically measured pedophilic interest than would other incest offenders against children (e.g., grandfathers, uncles, stepfathers). The prediction was not supported. A testable alternative hypothesis to explain biological father incest is presented and the importance of assessing pedophilic interest among incest offenders is discussed. PMID- 10369037 TI - Social cognition and the manic defense: attributions, selective attention, and self-schema in bipolar affective disorder. AB - Manic patients, depressed bipolar patients, and normal controls were compared on measures of social cognition. Manic patients showed a normal self-serving bias on the Attributional Style Questionnaire, but depressed patients attributed negative events more than positive events to self. On an implicit test of attributional style, both patient groups attributed negative events more than positive events to self. Both patient groups showed slowed color naming for depression-related but not euphoria-related words. Manic patients, like normal controls, endorsed mainly positive words as true of self but, like the depressed patients, recalled mainly negative words. Findings from the implicit tests indicate a common form of psychological organization in manic and depressed patients, whereas the contrasts between the scores on the implicit and explicit measures are consistent with the hypothesis of a manic defense. PMID- 10369038 TI - Perceptual biases in psychosis-prone individuals. AB - College students screened for psychosis-proneness using the Chapman scales were compared on 4 free-vision tasks that typically yield left-spatial-field biases. The tasks included 2 chimeric face tests, consisting of happy/neutral faces and male/female faces, and 2 nonface tasks, consisting of pairs of dot-filled or gradient-filled rectangles. Participants endorsing perceptual aberration items, magical ideation items, or both (n = 98) and control participants (n = 112) were left-biased on all tasks but gradients and were most biased on emotion faces; in contrast, i.e., social anhedonia participants (n = 40) displayed very little or no left-field biases. For all groups, task intercorrelations were greatest between the 2 face tasks and between the 2 nonface tasks. These findings suggest patterns of atypical perceptual asymmetry in psychosis-prone individuals. PMID- 10369039 TI - Physiological hyperarousal: construct validity of a central aspect of the tripartite model of depression and anxiety. AB - Physiological hyperarousal (PH) is an understudied component of the tripartite model of depression and anxiety. This study contributes to the literature on PH, the tripartite model, and anxiety and its disorders, using data from psychotherapy outpatients (n = 2,448), air force cadets (n = 1,335), and undergraduates (n = 284). Psychometrics and exploratory and confirmatory factor analyses showed that PH is a reliable, cohesive, discriminable, and valid construct. Compared with subjective anxiety, PH was more associated to panic versus mood disordered status, and to panic versus generalized anxiety disordered status. As hypothesized, an aspect of anxiety sensitivity (i.e., fear of body sensations) was particularly related to subjective anxiety in the presence of PH. Results support the PH construct as replicable, valid, and clinically important and support the utility of the tripartite and related models for understanding the relation of depressive and anxious syndromes. PMID- 10369041 TI - Attributions and affective reactions of family members and course of schizophrenia. AB - The authors tested an attribution-affect model of schizophrenic relapse attending to the role of families' positive affect (warmth) and negative affect (criticism). Coders listened to interviews of 40 family members taken from C. E. Vaughn, K. S. Synder, S. Jones, W. B. Freeman, and I. R. Falloon (1984) and rated their attributions of controllability for the symptoms and behaviors of their relatives with schizophrenia. For family members not designated as emotionally overinvolved, perceptions that their ill relatives' symptoms and behaviors were under the patients' control were related to family members' warmth and criticism and to patients' clinical outcomes. Of the affective reactions, only criticism predicted outcome. In addition, patients' use of street drugs was related to attributions, criticism, and outcome. Together these findings suggest that families' attributions and criticism are important in understanding the relationship between family factors and course of illness. PMID- 10369040 TI - Skin-conductance orienting deficits and increased alcoholism in schizotypal criminals. AB - This study tested the interaction hypothesis that a subgroup of criminals with schizotypal personality would show skin-conductance orienting deficits and increased alcoholism. In a prospective, longitudinal study of alcoholism in 134 males, schizotypy was assessed during adolescence, skin-conductance orienting was assessed at ages 18-20 years, and criminal offending and alcohol abuse were assessed at ages 30-33 years. A significant interaction between schizotypy and criminality indicated that schizotypal criminals were characterized by autonomic orienting deficits. Furthermore, the rate of alcoholism in schizotypal criminals (54.8%) was significantly higher than in criminals (23.8%), schizotypal noncriminals (13.9%), and comparisons (21.7%). It is argued that schizotypal criminals are a relatively distinct group and that prefrontal dysfunction may underlie both orienting deficits and alcoholism in this group. PMID- 10369042 TI - Neuropsychological characteristics and test behaviors of boys with early onset conduct problems. AB - School-age children and adolescents with conduct problems typically exhibit deficits in verbal IQ, language abilities, and executive functions. This study examined the extent to which this pattern was evident in a clinic group of preschool boys with early onset conduct problems who met criteria for oppositional defiant disorder (ODD) with and without attention deficit hyperactivity disorder (ADHD). A 2nd question focused on the strength of relation between clinic boys' uncooperative or inattentive test behaviors and their test performance. As expected, the clinic boys showed a neuropsychological profile highly similar to the one found in older conduct problem populations. Verbal tests distinguished clinic from matched comparison boys even after controlling for observers' ratings of disruptive behavior during testing. Clinic boys with ODD and ADHD had lower verbal and executive function scores than clinic boys with ODD alone. After general vocabulary knowledge and test behavior were controlled, clinic boys were found to have poorer vocabularies for describing affective states than comparison group boys. PMID- 10369043 TI - Dimensionality of posttraumatic stress disorder symptoms in children exposed to disaster: results from confirmatory factor analyses. AB - Factor analytic studies of trauma victims' posttraumatic stress disorder (PTSD) have offered conflicting hypotheses about how to conceptualize PTSD into symptom categories. The present study used confirmatory factor analyses of self-reported PTSD symptomatology from 5,664 child and adolescent victims of Hurricane Hugo to compare 10 models of PTSD dimensionality. PTSD was best represented by a 2nd order PTSD factor that manifests in 3 symptom clusters (Intrusion/Active Avoidance, Numbing/Passive Avoidance, and Arousal). This model was cross validated on 3 age groups (late childhood, early adolescence, and late adolescence), and results indicated factorial invariance across groups. PTSD symptoms varied in relative centrality to the underlying dimensions of PTSD, which differed in their relations with anxiety and degree of traumatic exposure. Implications for classification criteria and an empirically supported theory of PTSD are discussed. PMID- 10369044 TI - An event-related brain potential substrate of disturbed response monitoring in paranoid schizophrenic patients. AB - Response monitoring in schizophrenic patients and healthy controls was assessed by measuring performance and event-related brain potentials in the flanker priming task. Three visual-context conditions were construed: Flankers and targets pointed either into the same direction or into different directions. Stimuli without any response assignment were used as flankers in the neutral context condition. The schizophrenic patients were further subdivided into paranoid (n = 19) and nonparanoid (n = 10) patients and compared with healthy controls (n = 18). Performance scores revealed that the flankers induced a similar degree of distraction by visual context in all 3 groups. Although the schizophrenic patients showed normal error correction performance, the error negativity (NE) was significantly reduced in paranoid schizophrenic patients. The attenuation of the NE possibly reflects disturbed response monitoring in these patients. PMID- 10369045 TI - Physiologic reactivity to startling tones in women with posttraumatic stress disorder. AB - Autonomic and eyeblink reactivity to startling tones were investigated in women with histories of childhood sexual abuse (CSA). Twenty-one women with current posttraumatic stress disorder (PTSD), 23 with lifetime but not current PTSD, and 13 women who never had PTSD listened to 15 95-dB, 500-ms, 1000-Hz tones with a 0 ms rise time while heart rate (HR), skin conductance (SC), and orbicularis oculi electromyogram (EMG) responses were measured. Participants in the current and lifetime PTSD groups produced larger HR responses across tones and showed slower absolute habituation of SC response magnitude compared with the never PTSD group. EMG response magnitudes did not differ among groups. Women with CSA-related PTSD showed increased autonomic reactivity and slower habituation to high-intensity tones similar to that observed in primarily male, combat PTSD samples. This suggests that heightened autonomic responsivity to startling stimuli in PTSD is not gender or event specific. PMID- 10369046 TI - Are all psychopathic individuals low-anxious? AB - Theorists commonly assume that true or primary psychopathic individuals experience little anxiety or neurotic conflict. This study examined the relationship between psychopathy and anxiety in 104 Caucasian and 113 African American incarcerated men using the Psychopathy Checklist--Revised (PCL-R; R. D. Hare, 1991) and multiple self-report measures to tap diverse interpretations of the anxiety construct (i.e., neuroticism, traditional definitions of anxiety, and fear). Analyses involving zero-order, semipartial, and point-biserial correlations indicate that PCL-R psychopathy and the anxiety construct are essentially independent. These findings suggest that either (a) the traditional belief that all psychopathic individuals are low-anxious is incorrect or (b) the PCL-R is not an adequate measure of primary psychopathy. PMID- 10369047 TI - Eye-tracking dysfunction and birth-month weather in schizophrenia. AB - The prevalence of eye-tracking dysfunction (ETD) is significantly elevated in individuals with a diagnosis of schizophrenia and in their nonschizophrenic relatives, suggesting that ETD marks a familial (most likely genetic) risk factor for schizophrenia. Birth in a season with intemperate weather is also a widely reported risk factor for schizophrenia and is particularly marked for the subgroup with no family history of the disorder. This study examined how these two risk factors covaried in 78 patients with a Diagnostic and Statistical Manual of Mental Disorders (3rd ed., rev.; American Psychiatric Association, 1987) diagnosis of schizophrenia. Eye tracking and birth-month weather were independently assessed. As hypothesized, patients without ETD were significantly more likely to be born in months with intemperate weather (both hot and cold) than either patients with ETD or people in the general population. Etiologic factors associated with severe weather near birth may be important sources of nonfamilial schizophrenia. PMID- 10369048 TI - Clinical significance. AB - This article initiates the special section on clinical significance. Within a brief precis and overview, the 4 methodological articles and the integrative commentary of the special section are introduced. A call for the inclusion of the assessment of clinical significance in treatment evaluations is extended. PMID- 10369049 TI - Normative comparisons for the evaluation of clinical significance. AB - Normative comparisons are a procedure for evaluating the clinical significance of therapeutic interventions. This procedure, consisting of comparing data on treated individuals with that of normative individuals, is described, and a step by-step statistical methodology for conducting normative comparisons in the context of treatment-outcome research is presented. Four examples of the methodology are outlined in detail. Attention is paid to potential theoretical, statistical, and methodological challenges to the implementation of normative comparisons, as well as to the advantages of normative comparisons in providing evidence for the beneficial gains of treatment. PMID- 10369050 TI - Methods for defining and determining the clinical significance of treatment effects: description, application, and alternatives. AB - This article summarizes and scrutinizes the growth of the development of clinically relevant and psychometrically sound approaches for determining the clinical significance of treatment effects in mental health research by tracing its evolution, by examining modifications in the method, and by discussing representative applications. Future directions for this methodology are proposed. PMID- 10369051 TI - Assessing social validity in clinical treatment research: issues and procedures. AB - Social validity is a term coined by behavior analysts to refer to the social importance and acceptability of treatment goals, procedures, and outcomes. This article discusses dimensions of social validity, methods used to evaluate various aspects of social validity, and the applicability of these concepts and methods in clinical treatment research. PMID- 10369052 TI - Quality of life: expanding the scope of clinical significance. AB - Clinical researchers have turned their attention to quality of life assessment as a means of broadening the evaluation of treatment outcomes. This article examines conceptual and methodological issues related to the use of quality of life measures in mental health. These include the lack of a good operational definition of the construct, the use of subjective versus objective quality of life indicators, and the nature of the relationship between symptoms and quality of life judgments. Of special concern is the ability of quality of life measures to detect treatment-related changes. The authors review the application of quality of life assessment across diverse patient groups and therapies and provide recommendations for developing comprehensive, psychometrically sophisticated quality of life measures. PMID- 10369053 TI - The meanings and measurement of clinical significance. AB - The previous articles in this special section make the case for the importance of evaluating the clinical significance of therapeutic change, present key measures and innovative ways in which they are applied, and more generally provide important guidelines for evaluating therapeutic change. Fundamental issues raised by the concept of clinical significance and the methods discussed in the previous articles serve as the basis of the present comments. Salient among these issues are ambiguities regarding the meaning of current measures of clinical significance, the importance of relating assessment of clinical significance to the goals of therapy, and evaluation of the construct(s) that clinical significance reflects. Research directions that are discussed include developing a typology of therapy goals, evaluating cutoff scores and thresholds for clinical significance, and attending to social as well as clinical impact of treatment. PMID- 10369054 TI - Till violence does us part: the differing roles of communication and aggression in predicting adverse marital outcomes. AB - Measures of communication and aggression, taken from 56 newlywed couples, were used to predict marital outcomes 4 years later. Aggression discriminated between separated or divorced couples and those who remained married. In contrast, communication discriminated between couples who were maritally satisfied after 4 years and those who were married but dissatisfied. Thus, over the 1st 4 years of marriage, marital satisfaction and dissolution appear to be predicted by separate factors. These results remained unchanged after controlling for initial marital satisfaction. Additional analyses showed that 68% of the marriages could be accurately classified into their outcome groups using communication and aggression variables. These results help to integrate prior marital research on communication and aggression, and they suggest that it may be necessary to focus on both factors in efforts to strengthen marriages and prevent divorce. PMID- 10369055 TI - Spousal negative responses to cancer patients: the role of social restriction, spouse mood, and relationship satisfaction. AB - This cross-sectional study examined perceptions of spousal negative behaviors in 219 cancer patients. A mediational model was proposed to explain why a spouse might respond negatively to an ill partner because of greater restriction on activities as functional impairment increases. A moderating effect of the spouse's marital satisfaction assessed 3 months before other study measures was also proposed. Results provided support for the proposed model. The relationship between increasing patient functional impairment and spousal negative behaviors was medicated by greater restriction in spouse activities, as well as spousal negative mood. A marginally significant moderating effect for marital satisfaction was found. Although these results must be replicated with a prospective study, the findings begin to shed light on why spouses might respond in negative ways to an ill partner. PMID- 10369056 TI - The socioeconomic impact of interpersonal violence on women. AB - Prospective data from a nationally representative sample of women were used to examine 4 objective indexes of social adjustment following direct, interpersonal crime. Household income, marital status, employment, and education level were evaluated as risk factors for and outcomes of victimization. Data were collected in 3 waves at 1-year intervals, and 2,863 women completed all 3 waves. Results indicate that women experience increased risk for victimization when income is below poverty level and when newly divorced. Further, victimization appears to increase women's risk for unemployment, reduced income, and divorce. The cyclical nature of victimization is discussed. PMID- 10369057 TI - Sexually and physically abused foster care children and posttraumatic stress disorder. AB - Considerable debate exists regarding the possible relationship between child abuse and posttraumatic stress disorder (PTSD). In this study, 3 groups of foster care children were compared. The groups included 50 sexually abused, 50 physically abused, and 50 nonabused foster care children. Participants completed the Child Post-Traumatic Stress Reaction Index, the Childhood PTSD Interview, and the Modified Stroop Procedure (MSP), which included sexual abuse and nonsexual abuse stimuli. The MSP has not been previously used in child abuse research. Results indicated that sexually and physically abused children demonstrated PTSD at a high level. The MSP discriminated between the sexually abused children with PTSD and those without PTSD. Responses to the MSP sexual abuse stimuli resulted in significantly longer color-naming times than responses to nonsexual abuse stimuli. Preadolescents demonstrated more severe PTSD than early adolescent children. PMID- 10369058 TI - Positive and negative affectivity in children: confirmatory factor analysis of a two-factor model and its relation to symptoms of anxiety and depression. AB - The positive affect (PA) and negative affect (NA) framework that is embodied in the tripartite model of anxiety and depression has proved useful with adult populations; however, there is as yet little investigation with children concerning either the measurement of PA and NA or the relation between PA and NA and levels of adjustment. A confirmatory factor analysis was used in this study to examine the structure of self-reported affect and its relation to depressive and anxious symptoms in school children (4th to 11th grade). Results supported a 2-factor orthogonal model that was invariant across age and sex. Support for the expected pattern of relations between NA and PA with symptoms of depression and anxiety was strong for the older sample (M = 14.2 years) but weaker for the younger sample (M = 10.3 years). Results also provide preliminary support for the reliability and validity of the Positive and Negative Affect Schedule for children. PMID- 10369059 TI - Conscious and nonconscious African American stereotypes: impact on first impression and diagnostic ratings by therapists. AB - Sixty therapists randomly assigned to 1 of 2 priming conditions were primed with African American stereotypes or neutral words using 80-ms flash words on a computer screen. This procedure may activate information processing outside of conscious awareness. After this task, participants were exposed to a brief vignette introducing Mr. X, a patient referred for treatment, and then were asked to rate Mr. X on various dimensions. Results indicate that participants primed with stereotype words rated Mr. X significantly less favorably on hostility related attributes and significantly more favorably on hostility-unrelated attributes than did participants primed with neutral words. The findings suggest that therapists can be affected by African American stereotypes in ways that produce negative or positive first impressions depending on the nature of the attribute that is rated. PMID- 10369060 TI - Personality and substance use disorders: II. Alcoholism versus drug use disorders. AB - The relationship between personality and substance use disorders was investigated in a community-based sample of 638 individuals who were alcoholic and/or had a drug use disorder, and 1,530 individuals who did not have a substance use disorder. Personality was assessed by the Multidimensional Personality Questionnaire; substance use diagnoses were based on standard criteria as assessed by interview. Data were analyzed using a 3-factor (Gender x Alcoholism x Drug Use Disorder) multivariate analysis of variance. The significant alcoholism main effect was associated primarily with negative emotionality, whereas the significant drug use disorder main effect was associated primarily with constraint. No significant interactions with gender were observed. These findings suggest that the elevated levels of behavioral disinhibition observed with alcoholic individuals may be attributable to a subset of alcoholic individuals who also abuse drugs other than alcohol. PMID- 10369061 TI - Occult sleep apnea in a recruited sample of older adults with insomnia. AB - Although costly polysomnography (PSG) is not routinely performed with people with insomnia, it may be more necessary with recruited older adults with insomnia because this population may pose a greater risk of veiled sleep disorders compared with younger age groups and with referred samples. The present PSG screening of a recruited sample of older adults with insomnia found a 29%-43% rate of undiagnosed sleep apnea (SA), depending on whether an apnea-hypopnea index of 15 or 5 was used, after interviews had already screened out obvious cases of SA. Also, PSGs revealed a 4% rate of occult periodic limb movements. A discriminant analysis identified overweight men reporting dry mouth at highest risk for occult SA, with an apnea-versus-insomnia classification success rate of 78%. Using PSG evaluations in research on insomnia in recruited older adults is requisite to preclude substantial representation of occult SA. PMID- 10369062 TI - Chronic and discrete stress as predictors of children's adjustment. AB - Diabetes, as a chronic stressor, and negative life events (NLEs), as a discrete stressor, were related to children's behavioral adjustment, along with moderating effects of the family environment. Diabetes and NLEs predicted both higher internalizing (INT) and externalizing (EXT) behavior problems, suggestive of nonspecific distress. Higher family conflict and lower cohesion each predicted more behavior problems (INT-EXT). However, conflict was the sole moderator of the stressors. Higher family conflict and diabetes exacerbated children's EXT behavior problems, with clinically elevated scores. Higher family conflict and higher NLEs resulted in clinically elevated INT-EXT behaviors. Conversely, low family conflict protected children's behavioral functioning from the stressors. Family cohesion was the sole predictor of children's social competencies but did not moderate the stressors. PMID- 10369063 TI - Continuing care for cocaine dependence: comprehensive 2-year outcomes. AB - This report presents 2-year outcome data from an outpatient continuing care study in which cocaine-dependent patients (N = 132) were randomly assigned to either standard group counseling (STND) or individualized relapse prevention (RP). Data on cocaine outcomes during the 6-month treatment phase of the study were presented in an earlier report (J. R. McKay, A. I. Alterman, J. S. Cacciola, M. R. Rutherford, & C. P. O'Brien, 1997). In the present report, a continuing care condition main effect was obtained on only 1 of 8 outcome variables examined. However, patients who endorsed a goal of absolute abstinence on entering continuing care had better cocaine use outcomes in RP than in STND, whereas the opposite was the case for those with less stringent abstinence goals. In addition, patients with current cocaine or alcohol dependence on entering continuing care who received RP had better cocaine use outcomes in Months 1-6 and better alcohol use outcomes in Months 13-24 than those in STND. PMID- 10369064 TI - A randomized trial comparing day and residential drug abuse treatment: 18-month outcomes. AB - Extending an earlier report of 6-month outcomes, this study reports 12- and 18 month follow-up data for clients (N = 188) entering a therapeutic community drug treatment program who were randomly assigned to day or residential treatment conditions. Outcomes included Addiction Severity Index composite scores and measures of depression, psychiatric symptoms, and social support. Both groups showed significant change over time. The pattern of change indicated decreased problem severity in the 1st 6 months and then maintenance of lowered problem severity. Comparisons between groups indicated greater improvement for residential treatment clients on social problems and psychiatric symptoms but no differences on the remaining outcomes. Although residential treatment may offer some specific advantages, the conclusion here is that improvement among day treatment clients was not significantly different from that of residential treatment clients. PMID- 10369065 TI - Coping self-talk and cognitive interference in anxious children. AB - The present study addressed the as-yet-unresolved issue of whether coping self talk facilitates or interferes with effective task performance. Tests of the relationship between coping cognition and task performance are reported when potentially confounding relationships of negative cognition and task performance are controlled. The results indicate that coping self-talk of high-anxious children was positively correlated with negative thoughts but did not contribute significantly to performance. Implications for the functional value of coping self-talk are discussed. PMID- 10369066 TI - Antibody dependent cellular phagocytosis (ADCP) and antibody dependent cellular cytotoxicity (ADCC) of breast cancer cells mediated by bispecific antibody, MDX 210. AB - BACKGROUND: MDX-210 is a bispecific antibody (BsAb) with specificity for both the proto-oncogene product of HER-2/neu (c-erbB-2) and FcgammaRI (CD64). HER-2/neu is overexpressed in malignant tissue of approximately 30% of patients with breast cancer, and FcgammaRI is expressed on human monocytes, macrophages, and IFN-gamma activated granulocytes. We investigated phagocytosis and cytolysis of cultured human breast cancer cells by human monocyte-derived macrophages (MDM) mediated by BsAb MDX-210, its partially humanized derivative (MDX-H210), and its parent MoAb 520C9 (anti-HER-2/neu) under various conditions. MATERIALS AND METHODS: Purified monocytes were cultured with GM-CSF, M-CSF, or no cytokine for five or six days. Antibody dependent cellular phagocytosis (ADCP) and cytolysis (ADCC) assays were performed with the MDM and HER-2/neu positive target cells (SK-BR-3). ADCP was measured by two-color fluorescence flow cytometry using PKH2 (green fluorescent dye) and phycoerythrin-conjugated (red) monoclonal antibodies (MoAb) against human CD14 and CD11b. ADCC was measured with a non-radioactive LDH detection kit. RESULTS: Both BsAb MDX-210 (via FcgammaRI) and MoAb 520C9 (mouse IgG1, via FcgammaRII) mediated similar levels of ADCP and ADCC. ADCP mediated by BsAb MDX H210 was identical to that mediated by BsAb MDX-210. Confocal microscopy demonstrated that dual-labeled cells represented true phagocytosis. Both ADCP and ADCC were higher when MDM were pre-incubated with GM-CSF than when incubated with M-CSF. CONCLUSIONS: BsAb MDX-210 is as active in vitro as the parent MoAb 520C9 in inducing both phagocytosis and cytolysis of MDM. MDX-210 and its partially humanized derivative, MDX-H210, mediated similar levels of ADCP. GM-CSF appears to superior to M-CSF in inducing MDM-mediated ADCC and ADCP. These studies support the ongoing clinical investigations of BsAb MDX-210 and its partially humanized derivative. PMID- 10369067 TI - Comparative analysis of breast cancer recurrence risk for patients receiving or not receiving adjuvant cyclophosphamide, methotrexate, fluorouracil (CMF). Data supporting the occurrence of 'cures'. AB - PURPOSE: To comparatively analyse the risk of recurrence at given times after surgery for breast cancer patients receiving or not receiving adjuvant CMF. PATIENTS AND METHODS: A total of 1452 node positive patients, who entered controlled clinical trials carried out at the Milan Cancer Institute and underwent radical or modified radical mastectomy for operable breast cancer, were examined. In 575 cases no further treatment was performed, whereas 877 pts were given 6 or 12 courses of adjuvant Cyclophosphamide, Methotrexate, Fluorouracil (CMF). The recurrence risk was estimated by the event-specific hazard rate for first failure and distant metastases, and, following Efron, hazard rates were fitted by logistic regression models. RESULTS: The hazard rate for first failure and distant metastases showed a double peaked pattern for both treated patients and controls, with a first major peak at about 18-24 months from surgery (early metastases), a second minor peak at the 5th-6th year, and a tapered plateau-like tail extending over 10 years from surgery (late metastases). As expected, the recurrence risk of CMF treated patients was lower than the corresponding risk of patients undergoing surgery only. However, the difference was highly evident for early recurrences, while it declined and disappeared afterwards. CONCLUSION: Our findings confirm previous reports on patients not receiving adjuvant chemotherapy, suggesting that the recurrence risk for operable breast cancer has a multipeak pattern. As far as CMF treated patients are concerned, the unchanged peak timing together with the early recurrence risk reduction in comparison to controls are much more consistent with the real nonappearance of some early recurrences (putatively 'cured' patients) than with the delay in their manifestation. As late relapsing patients seem to have at most marginal benefits from adjuvant CMF, ways to recognize patients doomed to have late recurrence and new ways for treating micrometastases resulting in late recurrences are required. PMID- 10369068 TI - Pattern of distribution of cells positive for estrogen receptor alpha and progesterone receptor in relation to proliferating cells in the mammary gland. AB - Since cell proliferation is indispensable for the growth and development of the breast, and estrogens are considered to play a major role in promoting cell proliferation, while progesterone influences its differentiation, the present work was designed with the purpose of verifying the relationship between cells containing steroid hormone receptors and proliferating cells in the normal human breast. Twelve breast samples were analyzed for their content of lobules type 1 (Lob1), Lob2, Lob3, and Lob4, and the number of cells containing estrogen receptor alpha (ER-alpha), progesterone receptor (PgR), or expressing Ki67 antibody was determined by double immunocytochemical technique with specific antibodies. The highest percentage of ER-alpha, PgR, and Ki67 positive cells was found in Lob1, with a progressive reduction in the more differentiated Lob2 and Lob3. ER-alpha and PgR positive cells were found exclusively in the breast epithelium and were negative for Ki67, while cells positive for Ki67 did not express receptors. These findings were compared with the distribution of ER-alpha and PgR in the autoradiographs of mammary gland of young virgin rats inoculated with 3H-thymidine for determination of the DNA labeling index (DNA-LI). Both the DNA-LI and the percentage of ER-alpha and PgR positive cells were maximal in the epithelium of terminal end buds, and these values were reduced in alveolar buds and lobules. ER-alpha and PgR positive cells did not proliferate, and those cells that had incorporated 3H-thymidine were negative for both receptors. Our results led us to conclude that the content of ER-alpha and PgR in the normal mammary tissue varies with the degree of lobular development, in parallel with cell proliferation. However, the expression of receptors occurs in cells other than the proliferating cells, indicating that they represent at least two separate cell populations. These findings open new avenues towards the understanding of the mechanisms through which estrogens and progesterone affect the proliferative activity of breast epithelial cells, and their role in the initiation of the cascade of events that leads a normal cell to cancer. PMID- 10369069 TI - Increased AP-1 activity in drug resistant human breast cancer MCF-7 cells. AB - The expression, DNA binding, and transactivating activity of activator protein 1 (AP-1) was examined in a series of multidrug resistant (MDR) MCF-7 human breast cancer cells that have increasing levels of MDR1 gene expression. We observed an increase in the amount of both c-jun and c-fos mRNA in cells with 12-, 65-, or 200-fold higher resistance to adriamycin when compared to drug-sensitive MCF-7 wild type (WT) cells. Electrophoretic mobility shift assays (EMSA) demonstrated an increase in the DNA binding activity of an AP-1 complex in nuclear extracts from MDR MCF-7 cells when compared to extracts from WT cells. We observed a proportional increase in luciferase expression from a reporter vector containing consensus AP-1 binding sites in transiently transfected MDR cells when compared to WT cells, indicating that AP-1 mediated gene expression is increased in drug resistant MCF-7 cells. Since the MDR1 promoter contains a putative AP-1 binding site, we used EMSA to examine AP-1 binding activity to an oligonucleotide probe that contained the relevant MDR1 promoter sequences (-123 to -108). Nuclear extracts from resistant MCF-7 cells displayed an increased level of DNA binding of Jun/Jun dimers to the probe, indicating that AP-1 was capable of binding to this promoter site. A luciferase reporter construct containing triplicate copies of the MDR1 promoter sequence was expressed at higher levels in transiently transfected MDR cells when compared to expression in WT cells. Co-transfection of WT cells with a c-jun expression vector and either of the AP-1 luciferase constructs demonstrated that c-jun could activate gene expression from both the consensus and the MDR1 AP-1 sites in a dose dependent manner. In addition, RT-PCR and western blot analysis showed that levels of MDR1 mRNA and Pgp were increased in c-jun transfected WT cells. Taken together, these data indicate that increased AP-1 activity may be an important mediator of MDR by regulating the expression of MDR1. PMID- 10369070 TI - Diet and risk for breast cancer recurrence and survival. AB - Dietary factors may influence the risk for breast cancer and also the prognosis following diagnosis and treatment. The aim of this study was to assess whether self-reported prediagnosis diet or other patient factors associated with breast cancer incidence were predictive of recurrence and survival. Patients (n = 149) diagnosed with primary breast cancer between 1989 and 1991 were followed for five or more years. Total energy (hazard ratio (HR) = 1.58, 95%, confidence interval (CI) = 1.05, 2.38) as well as total (HR = 1.46, 95% CI = 1.05, 2.01), saturated (HR = 1.79, 95% CI = 1.05, 3.04), and monounsaturated (HR = 1.65, 95% CI = 1.09, 2.49) fat intakes were associated with increased risk, and energy-adjusted bread and cereal consumption (HR = 0.55, 95% CI = 0.33, 0.93) with decreased risk of recurrence. Both total energy (HR = 1.58, 95% CI = 1.03, 2.43) and polyunsaturated fat (HR = 1.84, 95% CI = 1.09, 3.13) intakes were associated with an increased risk of death. All associations between dietary fat and recurrence and survival attenuated following energy adjustment. Oral contraceptive use (HR = 1.28, 95% CI = 1.03, 1.60), lymph node positive status (HR = 2.36, 95% CI = 1.01, 5.49), and tumor stage (HR = 2.22, 95% CI = 1.02, 4.81) were associated with increased risk of recurrence. Tumor stage (HR = 4.96, 95% CI = 1.86, 13.23), lymph node positive status (HR = 3.31, 95% CI = 1.38, 7.95), and estrogen receptor negative status (HR = 2.46, 95% CI = 1.02, 5.94) were associated with increased risk, and arm muscle circumference (HR = 0.27, 95% CI = 0.09, 0.86) and mammographic utilization (HR = 0.77, 95% CI = 0.61, 0.98) with decreased risk of death. Higher levels of energy, fat intakes, and selected patient characteristics (particularly disease stage and anthropometric indicators of adiposity) appear to increase risk of recurrence and/or shortened survival following the diagnosis of breast cancer. PMID- 10369071 TI - Dose-dependent effect of tamoxifen therapy on endometrial pathologies in postmenopausal breast cancer patients. AB - To assess whether a higher cumulative tamoxifen dose is associated with increased incidence of various types of endometrial pathologies, we compared cumulative dose of tamoxifen treatment as well as demographic characteristics, risk factors for endometrial cancer, transvaginal ultrasonographic endometrial thickness, and various treatments for the primary breast cancer between 159 postmenopausal breast cancer tamoxifen-treated patients without endometrial pathologies (group I) and 67 similar patients with endometrial pathologies (group II). A similar comparison was made between group I patients and similar patients with proliferative endometrium (group IIa), with endometrial hyperplasia (group IIb), with endometrial polyps (group IIc), and with endometrial cancer (group IId). Overall cumulative tamoxifen dose was significantly higher in group II as compared to group I (27.4+/-33.4 and 17.4+/-20.2, respectively; P<0.0252). Transvaginal ultrasonographic endometrial thickness was significantly higher in group II than in group I patients (16.3+/-11.3 mm and 12.1+/-6.3 mm, respectively; P<0.0147). The frequency of diabetes mellitus, of previous postmenopausal bleeding, and of previous exposure to hormone replacement therapy was significantly higher in group II than in group I patients (P<0.001, P<0.0001 and P<0.001, respectively). There were no significant differences in all parameters tested between group I, group IIa, group IIb, group IIc, and group IId. However, there was an obvious trend for higher cumulative tamoxifen dose in patients with benign endometrial pathologies as compared to those without endometrial pathologies or to those with endometrial cancer (Group I = 17.4+/ 20.2 g, group IIa = 22.5+/-18.5 g, group IIb = 28.1+/-20.3 g, group IIc = 31.4+/ 42.7 g and group IId = 10.4+/-12.6 g). Endometrial pathologies, except for endometrial cancer, are associated with a high cumulative dose of tamoxifen in postmenopausal breast cancer patients. PMID- 10369072 TI - Neo-adjuvant chemotherapy for operable breast cancer induces apoptosis. AB - The use of neo-adjuvant chemotherapy (often referred to as pre-operative or primary chemotherapy) represents a major change in the management of breast cancer as a systemic disease. Laboratory studies have shown that many anti-cancer agents with differing modes of action achieve cytotoxic effects by inducing apoptosis. In this study, we investigated the induction of apoptosis by neo adjuvant chemotherapy in human breast cancer. The aim was to determine whether a correlation existed between post chemotherapy apoptotic index (AI) and clinical response and patients' survival. Our results indicate that apoptosis is induced by neo-adjuvant chemotherapy and that the response is variable. Our data show that post chemotherapy AI correlated with clinical response and increased patient survival, including both relapse (disease) free survival and overall survival. Post-neo-adjuvant chemotherapy AI levels in primary breast cancer may possibly predict an individual patient's overall response. PMID- 10369073 TI - Tamoxifen and risk of large bowel cancer in women with breast cancer. AB - BACKGROUND: The increasingly consistent association between estrogen replacement therapy and colorectal cancer suggests that the anti-estrogen tamoxifen may also be associated with large bowel cancer incidence. METHODS: Women with new diagnoses of breast cancer were identified from the Surveillance Epidemiology and End Results (SEER) Program, a set of geographically defined, population based cancer registries representing approximately ten percent of the U.S. population. Of 85,411 women with local or regional breast cancer diagnosed from 1983-90, 14,984 women were reported to have received hormonal therapy and 70,427 were not known to have received hormonal therapy. Subsequent cancer diagnoses were identified in this cohort beginning 6 months after initial breast cancer diagnosis until death, or December 31, 1994. Multivariate Cox proportional hazards models were used to estimate the risk of developing colorectal cancer and other second cancers according to hormonal therapy use. RESULTS: Over the follow up period 793 colorectal, 2,648 contralateral breast, 506 endometrial, 250 ovarian, 98 gastric, and 1,765 other cancers were identified in the study cohort. While overall there was no association between hormonal therapy use and colorectal cancer (relative risk (RR) 1.09, 95% confidence interval (CI) 0.88 1.35), in the period five or more years after diagnosis, risk was increased significantly by about 50% (95% CI 1.00-2.15). As expected, based upon clinical trials data, cancers of the contralateral breast were significantly decreased, and cancers of the uterine endometrium were significantly increased. No other meaningful associations were observed. When women were excluded for whom hormonal therapy might represent therapy other than tamoxifen (premenopausal women and those who received chemotherapy), this did not meaningfully alter these estimates. CONCLUSIONS: The results of this large population based cohort study suggest that tamoxifen therapy may modestly increase risk of large bowel cancer in women, but only after 5 years following initiation of breast cancer therapy. PMID- 10369074 TI - Recognition of an epitope of a breast cancer antigen by human antibody. AB - A novel tumor-associated peptide epitope KASIFLK expressed preferentially by breast cancer cells was identified using an IgG antibody from a breast cancer patient. A cDNA library from a MCF-7 breast cancer cell line was screened to isolate three cDNA clones that were immunoreactive with this antibody. KASIFLK was located in clones 27 and 40, both of which were identical to the cDNA and protein sequence of retinoblastoma binding protein 1 (RBP1, 250-256). An affinity purified IgG antibody against the peptide epitope was completely absorbed by cytoplasmic extracts of MCF-7 cells. Immunohistochemical staining using this antibody revealed the antigen in MCF-7 cells and in 12 of 15 primary breast cancer tissues and 3 of 34 other cancer tissues, but in none of 6 normal breast tissues. Anti-KASIFLK antibody titers were significantly higher in sera of 55 breast cancer patients than in sera from 30 normal healthy donors (P>0.001). These results suggest that KASIFLK or its cross-reactive epitope is a breast cancer antigen and is immunogenic in humans. PMID- 10369075 TI - BRCA1 and BRCA2 in breast cancer. AB - Breast cancer, one of the most common serious malignancies affecting women, occurs in hereditary and sporadic forms. Hereditary breast cancer accounts for 5 10% of all cases and has some distinctive clinical features compared with sporadic breast cancer. The recently identified genes BRCA1 and BRCA2 appear to account for the majority of hereditary breast cancer in US and European populations. Both of these genes have already been localized and isolated; however, the exact functions of their proteins are not clear yet. The detection of LOH in the 17q21 and 13q12-q13 regions, where the BRCA1 and BRCA2 genes are located, indicates that BRCA1 and BRCA2 act as tumor suppressor genes. The list of identified germline mutations in BRCA1 and BRCA2 is still growing, and mutation carriers have a substantial lifetime risk of both breast and ovarian cancer. However, it is still undetermined whether BRCA1 and BRCA2 play similar important roles in sporadic breast cancer. This paper reviews the current advances in BRCA1/BRCA2 research: the structure of their genes and proteins, their mutation frequencies, their possible roles in both hereditary and sporadic breast cancers, and their functions in transcriptional regulation and DNA repair. PMID- 10369076 TI - IGF-I and retinoic acid regulate the distribution pattern of IGFBPs synthesized by the canine mammary tumor cell line CMT-U335. AB - Stromal-epithelial interactions modulate growth and development in normal and neoplastic mammary gland. The release of IGF binding proteins (IGFBPs) by the stromal compartment of the mammary gland may play a modulating role in the IGF mediated proliferation of mammary epithelium. Therefore, the IGFBP-expression pattern of the canine mammary tumor cell line U335 (CMT-U335), which has a mesenchymal phenotype, was determined. In addition, the effects of IGFs and all trans retinoic acid (RA) on DNA synthesis, and IGFBP secretion and distribution were examined. The IGFBPs secreted by CMT-U335 were characterized as IGFBP-2, -4, -5, and -6. Moreover, CMT-U335 appeared to be a suitable mammary mesenchymal cell line for study of the regulatory factors of IGFBP expression and the mechanism(s) involved. IGFs and RA enhanced IGFBP concentrations in cell-conditioned medium with IGF-I and RA having an additive effect. The IGF-I-stimulated DNA synthesis, however, was inhibited by RA. The difference between IGF-I and RA was an enhanced IGFBP-5 binding to the extracellular matrix (ECM) by RA, whereas IGF-I reduced binding to the ECM. Because high doses of insulin had no significant effects on IGFBP concentrations in the medium, it is concluded that IGF-I-induced changes in IGFBP concentrations are not mediated by type-IIGF receptors and may be the consequence of IGFBP redistribution. PMID- 10369077 TI - Risk factors for a decline in upper body function following treatment for early stage breast cancer. AB - PURPOSE: To identify risk factors for a decline in upper body function following treatment for early stage breast cancer. METHODS: We conducted a cross-sectional observational study of 213 women > 55 years of age newly diagnosed with early stage breast cancer interviewed three to five months following their definitive surgery. Patients were classified as having impaired upper body function related to their breast cancer treatment if: 1) they reported having no difficulty in performing any of three tasks requiring upper body function (pushing or pulling large objects; lifting objects weighing more than 10 pounds; and reaching or extending arms above shoulder level) prior to treatment, but reported that any of these tasks were somewhat or very difficult in the four weeks prior to interview, or 2) they reported that performing any of the three tasks requiring upper body function was somewhat difficult prior to treatment, but reported that any of these tasks were very difficult in the four weeks prior to interview. RESULTS: In multiple logistic regression models, both the extent and type of primary tumor therapy and cardiopulmonary comorbidity were significantly associated with a decline in upper body function following breast cancer treatment. CONCLUSION: Given the critical importance of upper body function in maintaining independent living, clinicians should consider the functional consequences of treatment when they discuss treatment options and post-operative care with older women who have early stage breast cancer. PMID- 10369078 TI - Methodological arguments for the necessity of randomized trials in high-dose chemotherapy for breast cancer. AB - In the past ten years high-dose chemotherapy with autologous haematopoietic stem cell support (HD-CT) has increasingly been used for breast cancer. But the vast majority of trials are small phase I/II studies showing until now not enough evidence that HD-CT is superior to conventional-dose chemotherapy (CD-CT). In contrast to this, the public perception of this treatment is different. Patients as well as physicians often uncritically believe in reports contrasting the positive results obtained in case series treated by HD-CT with those of historical control groups. This leads to the problem that many patients and also clinicians are not willing to participate in randomized trials on this topic. A critical assessment of current knowledge on the effectiveness of HD-CT in breast cancer is given. The problems related to the use of historical controls, in general, and especially in the setting of HD-CT are demonstrated. Using data of patients treated with CD-CT within trials of the German Breast Cancer Study Group (GBSG) it will be shown that results similarly favorable to those reported from patients treated with a high-dose regimen may be produced using quite simple selection mechanisms. Comparisons of patients treated with HD-CT with historical control groups of patients treated with CD-CT may be misleading. A valid treatment comparison is only possible by means of large randomized trials. Clinicians should participate in the ongoing trials and enter all eligible patients. PMID- 10369079 TI - Synergistic cooperation between c-Myc and Bcl-2 in lymph node progression of T1 human breast carcinomas. AB - The overexpression of Bcl-2, an anti-apoptotic oncogene, identifies human T1 breast cancer patients who have an increased risk of lymph-node metastasis. We examined in these patients (n = 142) whether the c-Myc oncogene influences metastatic progression in conjunction or not with Bcl-2 expression and the loss of apoptosis in tumors. The association between Bcl-2 and lymph-node metastasis was only significant when c-Myc was concomitantly expressed (chi2 test, p = 0.008). Moreover, very large associations (pOR = 6.4) between c-Myc and lymph node metastasis were observed among Bcl-2 positive tumors and tumors with loss of apoptosis (pOR = 8.4). In contrast, the metastatic advantage linked to Bcl-2 was decreased (pOR = 2) when c-Myc was not coexpressed. It is concluded that the synergism between Bcl-2 and c-Myc oncogenes may promote metastasis in breast tumors, linked to loss of apoptosis. PMID- 10369080 TI - Quality of life in women with recurrent breast cancer. AB - Although the psychosocial concomitants of early-stage breast cancer have been well-documented, the relationship between cancer recurrence and quality of life remains less clear. We conducted a prospective, longitudinal study in order to clarify the relationship between recurrent cancer and quality of life, and to determine predictors of quality of life following recurrence. Sixty-nine women with recurrent breast cancer completed questionnaires assessing multiple components of quality of life at three time points: prior to recurrence, immediately after the diagnosis of recurrence, and at follow-up 6 months later. Perceptions of overall quality of life, general health status, emotional, social, and physical functioning were poorer immediately following the diagnosis of recurrence than they had been prior to recurrence. These women also evidenced significant improvement in several domains of quality of life between initial recurrence and follow-up; nonetheless, most areas of quality of life were impaired compared to pre-recurrence. Self-reported physical symptoms were a strong predictor of post-recurrence ratings of overall quality of life. These data suggest that the recurrence of breast cancer is associated with significant changes in quality of life. Quality of life did not progressively deteriorate, however, attesting to the resilience of women coping with this major stressor. These data shed light on issues of potential importance to patients managing this serious illness and may have implications for health-care professionals working with this population. PMID- 10369081 TI - Reduced telomere DNA content is correlated with genomic instability and metastasis in invasive human breast carcinoma. AB - Telomere shortening leads to genomic instability and has been correlated with poor outcome in several types of cancer. A recently described, robust titration assay was used to quantify telomere DNA content in frozen and paraffin-embedded specimens of 49 invasive human breast carcinomas, including tumors with normal or abnormal contents of genomic DNA, which produced regional, distant, or local disease. Telomere DNA contents ranged from 53% to 370% of the content in a reference DNA purified from normal placenta. Tumors were divided into three groups of approximately equal size based on increasing telomere DNA content. All of 16 tumors in the group with the least telomere DNA (Group I), were aneuploid compared to 9/17 tumors in the group with the most telomere DNA (Group III). The Chi-square test for trend indicated that tumors with the least telomere DNA were significantly more likely to be aneuploid than tumors with the most telomere DNA (p < 0.002). Twelve of 14 tumors in Group I also produced metastatic disease compared to 8/15 tumors in Group III. The Fischer Exact Test indicated that tumors with the least telomere DNA were significantly more likely to be metastatic than tumors with the most telomere DNA (p < 0.05). There was no association between telomere DNA content and patients' age, tumors' size, grade, stage, or fraction of cells in S-phase. The correlation of reduced telomere DNA content with aneuploidy and metastasis, both of which are associated with poor outcome in invasive breast carcinoma, implies that telomere DNA content also could have prognostic value. PMID- 10369082 TI - Tumor flow in malignant breast tumors measured by Doppler ultrasound: an independent predictor of survival. AB - The purpose of this study was to investigate tumor blood flow in breast cancers with regard to its impact on the overall survival of patients. Tumor blood flow was assessed in seventy-four patients with primary breast cancer by the use of color-coded Doppler ultrasound techniques. Preoperatively obtained Doppler frequency spectra were analyzed for peak systolic flow velocity (Vmax). Color Doppler signals were detected in 71 (96%) of the breast tumors. Out of 74 patients, 17 experienced a relapse or distant metastasis, and 15 women had died due to breast cancer at the time of data analysis. The mean Vmax of the patients who had died was 0.27 m s(-1), whereas survivors showed a mean Vmax of 0.16 m s( 1) (p = 0.01). Vmax, nodal status, and progesterone receptor status remained the only significant factors of overall survival in the multivariate model, whereas tumor size, tumor grade, and estrogen receptor status failed to retain prognostic significance. Moreover, Vmax was identified as the most important prognostic marker for survival in our series. The five-year-survival was 82.3% in Vmax < or = 0.25 m s(-1) patients versus 36.6% in women with tumor flow greater than 0.25 m s(-1). Patients with Vmax > 0.25 m s(-1) experienced a 4.33-fold increased risk of death secondary to the underlying disease. In summary, our data showed that tumor blood flow velocity measured by ultrasonography is an independent prognostic factor of survival in breast cancer patients. Furthermore, tumor flow velocity allows identification of patients at very high risk of death due to breast cancer. Large scale clinical trials should evaluate the clinical usefulness and future impact of this procedure for adjuvant treatment decisions. PMID- 10369083 TI - Prognosis, treatment, and recurrence of breast cancer for women attending or not attending the Screening Mammography Program of British Columbia. AB - Breast cancer screening programs have been initiated in many countries in the past decade. To determine the impact of the Screening Mammography Program of British Columbia (SMPBC), disease and treatment outcomes for women with breast cancer diagnosed in BC between 1989 and 1996 were compared on the basis of attendance at the SMPBC. An SMPBC attender was a women diagnosed with breast cancer within three years of an SMPBC screen, regardless whether the cancer was detected as a result of that screen. Of the 13,636 women aged 40-89 years diagnosed with breast cancer in BC during the study period, 2,647 (19.4%) were SMPBC attenders. 73.5% of SMPBC attenders (N = 1,946) and 74.2% of non-attenders (N = 8,149) were referred to the BC Cancer Agency and had pathology, staging, treatment, and outcome information available. SMPBC attenders compared with non attenders were more likely to have in situ disease alone, and those with invasive cancers had smaller tumors which were less likely to have grade III histology and less likely to have spread to axillary lymph nodes (all P < 0.001). SMPBC attenders were more likely to be treated with breast conservation and less likely to receive adjuvant chemotherapy or tamoxifen (P < 0.001). Log-rank tests showed local (P = 0.017), distant (P < 0.001), and overall (P < 0.001) disease-free survival were better for SMPBC attenders. These favorable surrogate endpoints suggest that the benefits of breast screening as demonstrated by randomized trials can be translated into community practice by an organized breast screening program. PMID- 10369084 TI - Analyzing prognostic factors in breast cancer using a multistate model. AB - In breast cancer clinical research, an important goal is to analyze how factors are seen to affect the disease process. Meanwhile, the disease progression is not fully modelled using standard analysis since transitions between intermediate events such as local-regional recurrences (LRR) or metachronous contralateral breast cancer (MCBC) are not considered. In the present study, the progression of disease was modelled using a multistate model. By this approach, we assessed transitions during the course of the disease and studied prognostic factors for each transition. The model was applied to 6,185 patients with unilateral ductal invasive breast cancer, clinical stage I through III, treated between 1981 and 1988 at the Curie Institute. At first diagnosis, high clinical stage, high histological grade, positive lymph nodes, and age less than 40 years were associated with increased risks of LRR, metastases, or death. Except age, the same factors remained predictive for metastases or death following LRR. Chemotherapy for the first cancer was associated with a decreased risk for developing MCBC. As the time interval from diagnosis of the primary tumor to that of a local or contralateral recurrence increased, the risk of metastases or death decreased. Nodal status for the first tumor and clinical stage for the contralateral tumor increased the risk of metastases or death following MCBC. Conversely, the risk decreased for patients who received adjuvant hormone therapy following MCBC. In conclusion, the multistate model offers us a much more appropriate way to study prognostic factors for each transition in breast cancer disease. PMID- 10369085 TI - The effects of thromboxane synthase inhibition on reperfusion injury and endothelin-1,2 levels in allograft kidney transplantation in rats. AB - Thromboxane A2 is a proaggregative vasoconstrictor that is synthesized and released in reperfusion injury. We aimed to investigate the effects of thromboxane synthase inhibitor, UK 38485, on endothelin-1,2 (ET) response of the renal endothelium and lipid peroxidation and protein oxidation in the early period of kidney transplantation. Four groups (n=8 in group IV and n=10 in the others) [corrected] of Sprague-Dawley rats were designed as Group I (sham nephrectomy), Group II (autotransplantation), Group III (allotransplantation) and Group IV (allotransplantation group in which the allografts were perfused with UK 38485. All subjects underwent right nephrectomy after transplantation. The grafts were flushed with 4 ml of ice-cold Ringer's lactate and in Group IV 10 microg of UK 38485 was added into the solution for each kidney. In allotransplantation groups, the kidneys were harvested from allogeneic white Wistar albino rats. The kidney grafts were allowed 120 min of reperfusion after 40 min of cold ischemic period. ET-1,2 plasma concentrations in the renal vein blood and diene conjugates (DC), hydroxyalkanals (HAA), hydroxyalkenals (HAE) and malondialdehyde (MDA) levels as the products of lipid peroxidation, protein carbonyls and protein sulfhydryls as the indicators of protein oxidation were analyzed in kidney tissue. Plasma ET-1,2 concentrations increased significantly in Group II and Group III (P<0.01) when compared to Group I but decreased in Group IV in comparison with Group III (P<0.05). DC, HAA, HAE and MDA levels increased in Groups II and III (P<0.001). Significant protein oxidation occurred only in Group III (P<0.01). Perfusion of the allografts with UK 38485 prevented lipid peroxidation and protein oxidation in Group IV. Histopathological changes were mild in the last group. We concluded that, in kidney transplantation, local administration of UK 38485 has cytopreservative effects on the allografts and this effect can be related to ET-1,2 concentrations. PMID- 10369086 TI - Nitecapone inhibits myeloperoxidase in vitro and enhances functional performance after 8 h of ischemia in experimental heart transplantation. AB - Nitecapone (NC) has been shown to have beneficial effects on the functional recovery of rat hearts in Langendorff-preparation. The present study was executed to evaluate the effect of NC on preservation of grafts in heart transplantation and the role of NC in the inhibition of granulocyte infiltration. Donor hearts were perfused and stored at +4 degrees C for 8 h in either Ringer solution in the control-group (C-group, n = 26) or in NC (50 microM) added Ringer solution (NC group, n = 18). The heterotopic heart transplantation was performed. The rats in both groups were killed at either 10 min or 60 min after release of the aortic clamp and tissue samples were obtained for antioxidative capacity, myeloperoxidase activity, and lipid peroxidation measurements. In vitro studies were performed using sodium azide or nitecapone to inhibit myeloperoxidase (MPO) activity of isolated human leukocytes. A total of 61% of the grafts began to beat in the NC-group, compared to 46% in the control group. Using an arbitrary scale of functional performance, only 33% (4/12) of the grafts were classified as well functioning in the control group, compared to 82% (9/11) in the NC-group (P<0.05). MPO activity was equal in both groups after 10 min but significantly lower after 60 min in the NC-group as compared to the control group (P<0.05). In vitro studies demonstrated that NC inhibits 50% of purified MPO activity at a concentration of 10 microM. NC did not significantly affect lipid peroxidation or the preservation of endogenous antioxidants. Since NC inhibited myeloperoxidase both in vitro and in vivo, it seems that the positive effects of NC on graft preservation may be mediated via the inhibition of granulocyte infiltration. PMID- 10369087 TI - Adaptive hepatic changes in mild stenosis of the common bile duct in the rat. AB - Adaptive hepatic changes were investigated in rats with mild stenosis of the common bile duct and in sham-operated controls. The studies were performed 24 h and 7-12 days postoperatively. A continuous intravenous infusion of taurocholic acid at stepwise-increasing rates was performed to explore the responses to bile acid effects. During the infusion, bile flow and the outputs of bile acids, phospholipids, cholesterol, alkaline phosphatase and gamma glutamyl transpeptidase were studied. At the end of the infusion, hepatic morphometric measurements were performed. In other experimental sets, biliary excretions of horseradish peroxidase, a marker of microtubule-dependent vesicular transport in the hepatocyte, and sulphobromophthalein, a well-known organic anion model, were studied. In other rats, bile acid pool size and composition were determined by depletion of bile. The results in rats with mild stenosis maintained for 24 h showed a greater susceptibility to the toxicity of taurocholic acid, as revealed by the abrupt decrement in bile flow at high rates of infusion, and increased outputs of phospholipids and canalicular enzymes. Conversely, rats with mild stenosis maintained for 7-12 days showed decreased bile acid maximum secretory rate and biliary outputs of phospholipids and canalicular enzymes, as well as hepatocyte hypertrophy. These findings may explain the limited hepatic and systemic repercussion of experimental mild stenosis of the common bile duct and help us to understand the early stages of constriction of the common bile duct in man. PMID- 10369088 TI - Protective capacity of a IgM/IgA-enriched polyclonal immunoglobulin-G preparation in endotoxemia. AB - Animal experiments were carried out to investigate whether a protective effect can be achieved in endotoxemia by intravenous (i.v.) application of a polyclonal immunoglobulin preparation (IVIG-IgG/A/M) enriched with 12% IgM and 12% IgA. Following administration of IVIG-IgG/A/M (500 mg/kg), endotoxemia was induced by intraperitoneal inoculation of a sublethal dose (5x10(8) CFU/kg) of Escherichia coli (E. coli) and subsequent i.v. administration of an antimicrobial agent (Imipenem). Plasma endotoxin activity, IL-6 activity, mean arterial pressure, and skeletal muscle oxygen pressure (tpO2) were measured at regular intervals over a total observation period of 7 h. Prophylactic administration of IVIG-IgG/A/M was found to significantly attenuate (P<0.01) the antibiotic-induced increase in endotoxin activity as compared to the albumin control group. Limited endotoxemia in the IgG/A/M group was associated with reduced levels of circulating IL-6 (P<0.01). Both lipopolysaccharide-induced hypotension and depression of tissue oxygenation were attenuated (P<0.01) by pre-treatment with IVIG-IgG/A/M. The experimental results suggest that in endotoxemia the polyclonal immunoglobulin preparation has a prophylactic protective effect on the acute phase responses and reduces the cardiodepressant effects of E. coli septicaemia. PMID- 10369089 TI - Superoxide dismutase activity and the effect of N-methyl-D-aspartate antagonists on lipid peroxidation in the early phase of cold injury. AB - Free radicals, lipid peroxidation and excitatory amino acids have been implicated in the secondary mechanisms of traumatic brain injury. We used the cold injury model in rats to assess the endogenous activity of the protective enzyme superoxide dismutase (SOD) and the lipid peroxidation level in the contused tissue at an early phase of injury. Furthermore, we treated the rats with two different N-methyl-D-aspartate receptor antagonists, namely MK-801 and CPP, and evaluated their effect on lipid peroxidation in the contused tissue. Rats were divided into four groups: sham, control, treatment 1 and treatment 2 groups (n= 16 for each group). Thirty and 60 min after craniectomy or injury, tissue samples were removed. SOD activity didn't change in this period. However, lipid peroxidation in terms of malondialdehyde (MDA) amount showed a significant increase at 60 min. Fifteen minutes after injury, MK-801 (1 mg/kg), CPP (10 mg/kg) or saline (1 ml) were applied intraperitoneally in treatment 1, treatment 2 and the control groups. Treatment with MK-801 attenuated MDA levels, whereas treatment with CPP did not. The protective effect of MK-801 achieved statistical significance. These results demonstrate that SOD activity does not change in the early period of cold injury. Moreover, these results show that lipid peroxidation increases after 60 min of cold injury, and treatment with MK-801 15 min after injury can prevent this elevation. PMID- 10369090 TI - Prehension with the ipsilesional hand after unilateral brain damage. AB - Sensorimotor deficits in the hand ipsilateral to a brain lesion have been reported in different motor tasks. We evaluated performance of the ipsilesional hand in 12 patients with either left (LBD) or right brain damage (RBD) by kinematic analysis in order to precisely characterize possible deficits in the two components of prehension (transport and grasp). Both patient groups exhibited performance deficits in the main kinematic parameters, e.g., reduced velocity of the transport component and prolonged movement time. However, while LBD patients showed a more general slowing, RBD patients prolonged in particular the last phase of the movement toward the object. We suggest that relevant visuospatial representations and the adequate mapping of motor processes may be impaired after RBD. In contrast, LBD caused a more unspecific disturbance pattern, supporting the view that the precise parameterization of motor programs is impaired. Maximum grip aperture was normal in both patient groups. However, since aperture could be biased by slowed movement, the notion that the grasp component was preserved remains speculative. The patient's ability to scale the maximum velocity of the transport component to adapt to changes in movement amplitude and to scale the maximum hand aperture of the grasp component to adapt to object size was preserved in both groups. Thus both hemispheres can have competence for this scaling mechanism. PMID- 10369091 TI - Content-specific confabulation. AB - This report describes a person who confabulated following an anterior communicating artery aneurysm. His confabulation was limited to one very circumscribed area of his life and remained stable for twelve weeks, eventually improving with rehabilitation. It is argued that a content-specific confabulation of this nature challenges current theories of confabulation, and an integrative explanation is given based on previous findings by Burgess and Shallice (1996b) of the mechanisms of autobiographical recollection in healthy people. PMID- 10369092 TI - Crossed apraxia: implications for handedness. AB - Liepmann posited that right hand preference relates to left hemisphere dominance for learned skilled movements. Limb apraxia, impairment of skilled movement, typically occurs in individuals with left hemisphere (LH) lesions. The occurrence of apraxia in right-handed individuals following right-hemisphere lesions appears to refute Liepmann's hypothesis. We studied the apraxia of a right-handed man, RF, following a right frontal lesion to determine whether his apraxia paralleled the apraxia seen following LH lesions. Results of behavioral testing indicated that, like individuals with apraxia following left frontal lesions, RF was better at gesture recognition than gesture production which was significantly impaired across tasks. Kinematic motion analyses of movement linearity, planarity, and the coupling of temporospatial aspects of movements substantiated the parallel impairments in RF and patients with LH apraxia. The impairment seen in our patient with crossed apraxia provides evidence for the fractionation of systems underlying hand preference and skilled movement. PMID- 10369093 TI - A computational model of neglect dyslexia. AB - This paper presents a straightforward theory of neglect (a pre-attentive salience system selects objects' centers for the focus of an attentional "spotlight") as a computational model. This construction permits simulations of model "lesions" and allows checking unequivocally the model's implications. The current model can account for some of the common patterns of observations of neglect dyslexia: that errors increase with word length and that short words may be read as "too long". Most importantly, it also accounts for a dissociation between neglect dyslexia and neglect on line bisection tasks. A two stage model can account for the patterns of performance seen in many neglect subjects, but more important is the demonstration that a theory's implementation as a computational model permits unambiguous evaluation of the theory's implications. PMID- 10369094 TI - Mnestic block syndrome. AB - The case of a patient with largely preserved intelligence, but severe and persistent memory impairments is reported. FA, a 46-year-old patient with the diagnosis of prolonged depression was investigated repeatedly over a two year period with neuroradiological, neuropsychological, neuromonitoring and other methods. While no brain damage was detectable in FA, he manifested continued and severe anterograde and retrograde memory disorders together with an inhibition in his thinking processes. Otherwise, his intellectual capabilities were in the normal range, that is he was not pseudo-demented. Various approaches with drug treatment and psychotherapy failed to improve his condition. The condition is interpreted as 'mnestic block syndrome' and is considered to be related to an altered brain metabolism which may include changes in various transmitter and hormonal systems (GABA-agonists, glucocorticoids, acetylcholine). Whether depression contributes to this syndrome is uncertain from FA's cognitive performance, but may be a possibility. PMID- 10369095 TI - Sex related effect of unilateral brain lesions on the perception of the Mueller Lyer illusion. AB - The aim of this study was to investigate the effect of unilateral brain lesions on Mueller-Lyer (M-L) illusion in the two sexes. Patients with left hemisphere (LH) and right hemisphere (RH) damage and control subjects participated in the experiment. They inspected series of M-L patterns in which the shaft with out going fins was gradually shortened until it induced a perception opposite to the original illusion, that is, the shaft with out-going fins appeared to be shorter than the shaft with in-going fins. The subjects' task was to decide, on each trial, whether the variable shaft was longer or shorter than the other one. The point where the judgements changed from one category to the other was established using the Spearman distribution method for determining psychophysical thresholds, and was considered the measure of the strength of the illusion. The higher the value of the threshold, the stronger the illusion. Our results showed sex-related hemispheric asymmetry in subjects' susceptibility to the M-L illusion, i.e., both LH and RH lesions in females, but only RH lesions in males resulted in an increase of the strength of illusion. Moreover, males with LH lesion as well as controls partially corrected the illusory perception with practice, while both LH and RH damaged females and RH damaged males did not show this learning effect. PMID- 10369096 TI - Retrograde amnesia for world knowledge and preserved memory for autobiographic events. A case report. AB - A patient (PC) with severe and chronic retrograde amnesia for world knowledge (tested with famous events and famous faces), but unimpaired autobiographical memory is described. The 64-year-old man had traumatic brain injury four years prior to the present evaluation. Current brain imaging showed principally damage involving the infero-lateral prefrontal and the lateral temporal regions of the left-hemisphere. PC was of average intelligence, had no depression and only minor language problems, but manifested some additional anterograde memory deficits and performed subaverage in various frontal lobe-sensitive tests. Patient PC represents one of the very few cases with a preserved retrograde episodic and an impaired retrograde knowledge system, showing a dissociation between preserved retrieval of autobiographical events and amnesia for nonpersonal famous events. It is hypothesized that the sparing of autobiographical memories can be linked to the integrity of the right frontal and temporo-polar cortices. PMID- 10369097 TI - Quantitative analysis of cancellation tasks in neglect. AB - Patients with spatial neglect do not explore contralateral space effectively. Although cancellation tasks are used widely to assess this visual search deficit, their methods of analysis are not well established. We introduce logistic regression analyses for cancellation tasks in 7 patients with left neglect. We investigated the influences of spatial location, stimuli number, and target discriminability on the probability of canceling a target. As a group, neglect patients showed left and near neglect. They also explored and canceled targets further into contralateral space on arrays with fewer visual stimuli. Individual analyses revealed exceptions to these group patterns, such as two patients with far rather than near neglect. Only patients with relatively mild neglect canceled more targets when they were more easily discriminated from distracters. Logistic regression models accounted for 0.68 the variance in cancellation performances of the entire group. Shifting the unit of analysis from the proportion of targets canceled to the probability of detecting individual targets offers a powerful parametric method to analyze group and individual performances on cancellation tasks and can reveal functional dissociations in neglect. PMID- 10369098 TI - Spatial cognition in males with Fragile-X syndrome: evidence for a neuropsychological phenotype. AB - Spatial performance in a group of young Fragile-X syndrome males with FMR-1 full mutation was compared to a learning disabled control group comprising young Down's syndrome males and two control groups of mainstream schoolchildren. Performance was assessed across a wide range of spatial tasks including visuo construction, visuo-spatial memory, visuo-motor, and visuo-perception. The findings indicate a task-specific rather than global deficit in spatial performance in Fragile-X males with visuo-constructive and visuo-motor skills most vulnerable. Molecular analysis of the lymphocyte DNA found minimal evidence for a correlation between CGG expansion size and spatial performance, although tasks with a visuo-perceptual component correlated negatively with expansion size indicating that the further away the number of repeats are from the 200 threshold the poorer the performance. PMID- 10369099 TI - Gender affects word retrieval of certain categories in semantic fluency tasks. AB - Recent studies suggest that gender influences phonetically-cued fluency and some semantic memory tasks. In this study we analysed the effect of demographic variables on semantic fluency tasks. The semantic categories considered were: animals, fruits, tools and vehicles. The influence of age and education was common to all the categories considered and seems a general characteristic of the semantic fluency task. Gender had a significant effect only with fruits and tools, but a diverging role: females fared better with fruits and males with tools. We discuss whether the source of the gender effect should be located at the level of the semantic representation of each category or at the level of item recall in the short time (one minute) granted for the task. PMID- 10369100 TI - Pure topographical disorientation due to right posterior cingulate lesion. AB - We report an 82-year-old woman who developed pure topographical disorientation after a cerebral infarction involving the isthmus of the right posterior cingulate gyrus. She lost her way in new environments such as the hospital, but not in old ones such as her own house. She correctly identified familiar or unfamiliar landscapes and buildings by photographs. Her failure to memorize a new route likely resulted from a loss of directional memory over a wide area. We suggest that the right posterior cingulate gyrus contributes to memorizing a new route. PMID- 10369101 TI - The value of electron microscopy in the diagnosis and clinical management of lupus nephritis. AB - The diagnosis and clinical management of patients with lupus nephritis can be a challenge from a clinicopathologic point of view. Although the majority of patients that are biopsied already have either an established clinical diagnosis or a presumptive diagnosis of systemic lupus erythematosus, determination of the immunomorphologic characteristics, pattern, and distribution of renal involvement is important for clinical management. In a clear subset of these patients with lupus nephritis, electron microscopy plays a pivotal role in accurately characterizing the type of renal involvement and determining the degree of activity, providing useful and objective guides for patients' management. Ultrastructural evaluation can also be crucial in the initial diagnosis of patients with lupus who, at the time of biopsy, lack either diagnostic clinical manifestations and/or serologic markers, and are therefore clinically unsuspected. Electron microscopic evaluation also plays a significant role in the evaluation of renal dysfunction in transplant patients with lupus nephritis, helping to determine whether recurrence of the lupus has occurred in the renal allograft. There are some ultrastructural findings that, although not pathognomonic, in the proper clinico-pathologic context are very suggestive or even diagnostic of lupus nephritis. Correlating light, immunofluorescence, and electron microscopic findings within the clinical context of lupus nephritis cases is crucial for appropriate clinical management. In some of these patients, electron microscopy provides key information that cannot be otherwise obtained. PMID- 10369102 TI - The macrophage origin of the HIV-expressing multinucleated giant cells in hyperplastic tonsils and adenoids. AB - Replication and storage of virus are characteristic features of hyperplastic lymphoid tissues in HIV infection. In opportunistic infections, HIV is synthesized by phagocytic mononuclear and Langhans'-type multinucleated macrophages that coexpress the dendritic cell-associated S-100 and p55 antigens. However, similar cells in hyperplastic tonsils and adenoids from HIV+ individuals were alternatively identified as macrophages or, on the basis of the same S-100 and p55 staining, as dendritic cells. To consider establishing the role of these HIV-rich cells in HIV disease, it is important to reconcile this apparent discrepancy in identity. Hyperplastic tonsils and adenoid specimens were analyzed by HIV RNA in situ hybridization (ISH), light and transmission electron microscopy (TEM), and immunohistochemistry (IHC) (HIV Gag p24 protein, S-100, p55, CD68, HAM56, lysozyme, alpha-1-anti-trypsin, and alpha-1-anti-chymotrypsin). In HIV+ pediatric and adult surgical specimens (n = 11), the giant cells and their mononuclear counterpart were positive for both macrophage and p55 and S-100 IHC markers. In addition, TEM, p24 IHC, and ISH showed HIV expression by cells with typical features of macrophages. Furthermore, these cells were not unique to HIV+ specimens, being seen in 20% of hyperplastic T&A surgical specimens (n = 57) lacking HIV as well as in several types of granulomatous processes, such as sarcoidosis. These cells appear to represent an activated phenotype that can develop independent of HIV, but that may represent a viral host in HIV-infected individuals. Thus, the giant and mononuclear cells that produce striking amounts of HIV in tonsils and adenoids are of macrophage origin, yet, as in opportunistic infections, share dendritic cell-associated antigens, reflecting a common CD34+ bone marrow progenitor. PMID- 10369103 TI - The initial phase of myocardial reperfusion is not associated with aggravation of ischemic-induced ultrastructural alterations in isolated rat hearts exposed to prolonged global ischemia. AB - The present study focuses on the qualitative and sequential development of myocardial ultrastructural changes during the first 10 min of reperfusion in isolated rat hearts exposed to 60 min of global ischemia. The frequency of and the association between ultrastructural changes were examined by semiquantitative morphometry using the micrograph as unit. In each micrograph the subcellular components of the myocytes (sarcolemma, mitochondria, myofilaments and nucleus) and the endothelial cells were evaluated and graded as slightly, moderately, or severely altered. Ischemia alone induced moderate to severe ultrastructural alterations. The myocytes revealed sarcolemmal disattachment or rupture. The myocytic mitochondria had a clear matrix with abundant broken cristae and amorphous matrix densities. The myofilamental pattern was irregular or even disrupted, and most nuclei had reduced density and showed margination of chromatin. The endothelium showed vacuolization, rupture of the plasma membrane, and extracellular accumulation of cellular debris. During the first 2 min of reperfusion severe ultrastructural alterations were partly reversed. After 10 min of reperfusion both the frequency and grade of myocardial ultrastructural alternations were similar to that observed after ischemia. Cristal adhesions occurred predominately during reperfusion and were associated with moderately and severely altered myocytic mitochondrial alterations. In conclusion, the results showed that ischemic-induced ultrastructural alterations were transiently improved upon reperfusion. With exception of the development of cristal adhesions, the acute phase of reperfusion was not associated with additional ultrastructural changes in isolated buffer-perfused rat hearts exposed to prolonged ischemia. PMID- 10369104 TI - Glomerulopathic light chain-mesangial cell interactions modulate in vitro extracellular matrix remodeling and reproduce mesangiopathic findings documented in vivo. AB - Glomerulopathic light chains (LCs) are associated with two distinct mesangiopathies: AL (light-chain-related) amyloidosis and light-chain deposition disease (LCDD) with immunomorphologic features that are well documented in the literature. Even though both conditions are caused by monoclonal LCs, these entities differ dramatically in their morphologic expressions. In AL amyloidosis the mesangial matrix is replaced by amyloid fibrils, while in LCDD the matrix increases as a consequence of deposition of excess extracellular matrix (ECM). The immunomorphologic mesangial alterations observed in biopsy material are closely reproduced in vitro when mesangial cells grown on an artificial matrix are incubated with monoclonal light chains obtained from the urine of patients with either condition. This article summarizes previously reported data, reports new findings, and focuses on integrating all the available information on the subject. When mesangial cells are incubated with LCDD-LCs, production of ECM proteins (collagen IV, laminin, fibronectin, and tenascin) is increased, with maximum effect at 72 hours post LC treatment. A concomitant decrease in collagenase IV activity further accentuates the accumulation of mesangial matrix. These effects are mediated through transforming growth factor-beta (TGF-beta) activation. In contrast, when mesangial cells are incubated with Am-LCs, a decrease in ECM protein production and a stimulatory effect on collagenase IV is observed, which results in matrix degradation and facilitates amyloid deposition. The decreased TGF-beta documented in the literature in this setting precludes adequate matrix repair. These findings substantiate the morphologic alterations observed in renal biopsy specimens and in the in vitro model. Using this in vitro model, it is then possible to delineate the LC interactions with putative receptors at the mesangial cell surface that regulate mesangial cell pathobiologic responses and mesangial matrix homeostasis. PMID- 10369105 TI - Langerhans cell histiocytosis of the vulva: an ultrastructural study. AB - Langerhans cells histiocytosis (LCH) is a proliferative disorder of Langerhans cells. The lesions are normally characterized by infiltration of eosinophils, neutrophils, lymphocytes, plasma cells, and Langerhans cells. The specific cells of LCH contain Birbeck granules, express the phenotype of Langerhans cells but with markers fixed at an early stage of activation, and are functionally defective in antigen-presenting ability. The disease most often affects children; when it occurs in older patients, anal and groin involvement is quite common and vulvar lesions can be found in older females. The authors report a case of a 64 year-old woman with LCH of the vulva and diabetes insipidus. An immunohistochemical and ultrastructural study of the vulvar lesions showed an infiltrate in which antigenically and morphologically mature Langerhans cells, monocytoid cells, and cells with an intermediate phenotype between monocytes and Langerhans cells were concurrently observed. Although the clinical and histological aspects of LCH are well established, the pathogenetic mechanism of lesions is not yet known. The finding of an infiltrate composed by Langerhans cells and many putative precursors of these cells suggests the hypothesis of an in situ differentiation of Langerhans cells from immature monocytoid precursors. PMID- 10369106 TI - Small cell osteogenic sarcoma of the ribs: cytological, immunohistochemical, and ultrastructural study with literature review. AB - Small cell osteosarcoma (OS) is a rare variant of OS that is composed of small cells resembling those of Ewing's sarcoma (ES) with recognizable osteoid. This type of tumor often creates difficulty in making a diagnosis when tissue samples do not include osteoid. The frequent sites are long bones and until now there have been no reported cases arising in the ribs. A case is reported here of small cell OS occurring in the ribs of a 37-year-old female with its aspiration cytologic and electron microscopic characteristics. In the cytologic smear, the small round neoplastic cells were individually scattered or arranged in small nests. The nuclei were hyperchromatic and oval with no visible nucleoli. Ultrastructurally, the nuclei had a round or oval euchromatic chromatin pattern and occasional nucleoli. The scanty cytoplasm contained a small quantity of organelles including either tubular or dilated cisternae of RER, a few mitochondria, and free or polyribosomes. Other organelles were absent. Although the electron microscope sample of this case did not include bone mineral (hydroxy apatite), the electron microscopic features of the tumor cells were unique and useful for exclusion of other small round cell neoplasms. PMID- 10369107 TI - Serum amyloid A (SAA): a concise review of biology, assay methods and clinical usefulness. AB - Serum amyloid A (SAA) is a family of proteins encoded in a multigene complex. Acute phase isotypes SAA1 and SAA2 are synthesized in response to inflammatory cytokines. SAA and C-reactive protein (CRP) are now the most sensitive indicators for assessing inflammatory activity. In viral infection and kidney allograft rejection, SAA proved more useful than CRP. Development of convenient assay methods for SAA will facilitate its use in clinical laboratories. PMID- 10369108 TI - Serum cystatin C as an endogenous marker of the renal function--a review. AB - Since 1985, cystatin C has been suggested to be a marker of the renal function. Cystatin C is a proteinase inhibitor with a low molecular weight (M(r) = 13359). It is produced at a constant rate in all nucleated cells investigated to date, freely filtered in the renal glomeruli and reabsorbed and catabolised in the proximal tubules. The concentration of serum cystatin C is mainly determined by glomerular filtration, which makes cystatin C an endogenous marker of glomerular filtration rate (GFR). There are few data describing the influence of various factors on the production and elimination of cystatin C. Fully automated assays using particle-enhanced turbidimetry or particle-enhanced nephelometry are available and the assays are precise, rapid and usable in clinical routine practice. Reference intervals have been determined for cystatin C in adults and in children older than one year. It has been suggested that the same reference interval can be used in children older than one year and in adults without gender differences, on the assumption that the same method with the same standardisation is used. Several studies including adults and children with different renal diseases with various kidney function have suggested serum cystatin C to be a better marker of GFR than serum creatinine. PMID- 10369109 TI - Improved fluorescent PCR-based assay for sizing CGG repeats at the FRAXA locus. AB - Fragile X syndrome is the most frequent heritable genetic disease involving mental retardation and is usually caused by an expanded CGG repeat in the first exon of the FMR1 gene. Therefore, searching for CGG expansion at the FRAXA locus among the mentally retarded has become a routine investigation in neuro paediatric practice. Consequently, we have developed a fluorescent PCR-based assay for sizing repeats as an alternative to laborious and time-consuming Southern blot. The procedure utilises a reverse fluorescent labelled primer, and the Expand Long Template PCR system (Roche) with addition of dimethylsulfoxide and 7-deaza-dGTP It allows precise determination of the CGG repeat number in males and females for alleles from normal to premutation size range and detection of full mutations in males. We believe that this PCR protocol, allowing a high sample throughput, is useful for first-line screening among mentally retarded males, possibly complemented by Southern blot analysis to assess the methylation status of large mutated alleles. PMID- 10369110 TI - The influence of radiation and chemotherapy-related DNA strand breaks on carcinogenesis: an evaluation. AB - PURPOSE: DNA strand breaks are believed to induce carcinogenesis. This study was conducted to analyze induction and repair of irradiation- and chemotherapy related strand breaks in vitro. METHODS: Friend Leukemia cells were exposed to irradiation and various chemotherapeutic agents at different doses and concentrations. Occurrence of strand breaks was determined fluorometrically, measuring the rate of DNA unwinding immediately after exposure and 24 hours later. RESULTS: The amount of double-stranded DNA decreased significantly for irradiation, doxorubicin, dactinomycin and etoposide (p < or = 0.05, t-test). After 24 hours free of exposure, the persistent damage was detectable for all of these agents but not for irradiated cells, with DNA strand breaks being decreased for etoposide, unchanged for doxorubicin and increased for methotrexate as well as for dactinomycin. CONCLUSIONS: Severe DNA damage is induced by various chemotherapeutic agents and by irradiation. While repair of chemotherapy-related strand breaks may remain incomplete or prolonged for some chemotherapeutic agents, repair of radiation induced strand breaks is faster and more complete. Therefore chemotherapy-related carcinogenesis may partially be explained by prolonged persistence of DNA strand breaks. PMID- 10369111 TI - A simple HPLC method for monitoring mycophenolic acid and its glucuronidated metabolite in transplant recipients. AB - Mycophenolic acid (MPA) is nowadays in broad clinical use as a substitute for azathioprine. An immunoassay for MPA recently received approval for clinical applications. The high performance liquid chromatography (HPLC) assay for measuring MPA and its glucuronide conjugate (MPAG) we describe here is not only rapid and simple but also extremely sensitive at plasma levels obtained during standard immunosuppressive regimens. The determination of MPAG is possible without any change of the chromatographic conditions (detection wavelength of 214 nm, mobile phase: acetonitrile and 50 mmol/l o-phosphoric acid (50:50, V/V), run time: 15 min). The required equipment is a standard HPLC system including a simple UV-detector. Sample volume of 400 microl is required for both determinations. Detection limit is 0.25 micromol/l for MPA and 5 micromol/l for MPAG. Linearity is excellent for serial dilutions (0.5-25 micromol/l for MPA, 25 500 micromol/l for MPAG) and high accuracies favour the method described. More than 2000 plasma samples tested for MPA in patients after heart transplantation within one year and more than 500 samples for MPAG underline the clinical applicability of this assay. PMID- 10369112 TI - The activities of some glycosaminoglycan-degrading enzymes in the wall of the umbilical cord artery and their alteration in edema, proteinuria, hypertension (EPH)-gestosis. AB - Edema, proteinuria, hypertension (EPH)-gestosis is associated with a premature replacement of hyaluronic acid by sulphated glycosaminoglycans (GAGs), both in the umbilical cord arteries (UCAs) and in Wharton's jelly. This phenomenon may be considered as a sign of premature ageing of the umbilical cord tissues. The decrease in hyaluronic acid content in the UCA was found to be the result of reduced biosynthesis of this substance, whereas an increase in sulphated GAGs content is rather a result of slower degradation of newly synthesised GAGs. In this study the activities of GAGs-degrading enzymes in normal umbilical cord arteries and those taken from newborns delivered by mothers with EPH-gestosis were compared. It was found that EPH-gestosis results in a significant reduction in the activities of neutral endoglycosidases degrading most of the sulphated glycosaminoglycans (with the exception of heparan sulphate). The activities of exoglycosidases also decrease but to a lesser degree. These alterations are thought to be responsible for EPH-gestosis-associated accumulation of sulphated glycosaminoglycans in the extracellular matrix of the arterial wall. Such remodelling of the arterial wall may affect foetal blood circulation. The significance of these phenomena in the pathological mechanism of EPH-gestosis is discussed. PMID- 10369113 TI - Endothelin-1- and endothelin-receptors in lung biopsies of patients with pulmonary hypertension due to congenital heart disease. AB - Endothelin-1 (ET-1), with its vasoconstrictive and proliferation-stimulating effects, could play a role in the pathogenesis of primary pulmonary hypertension. We investigated the relationship between the ET-1 like immunoreactivity and the ET-receptor density, the grade of the pulmonary vasculopathy, and properties of the pulmonary circulation in patients with pulmonary hypertension due to congenital heart disease. Twenty-six patients with a median age of 1 year and 1 month (6 weeks - 17 years - 9 months) were assigned to group I (n = 15) with a pulmonary to systemic flow ratio (Qp/Qs) > or = 1.5 and a pulmonary to systemic resistance ratio (Rp/Rs) < or = 0.3 ("high flow - low resistance group") and to group II (n = 11) with a Qp/Qs < 1.5 and an Rp/Rs > 0.3 ("low flow - high resistance group"). Patients belonging to group II showed a higher ET(A)-receptor density in lung arteries (p < 0.05) and parenchyma (p < 0.01) than patients in group I. Patients with the highest ET-1 like immunoreactivity in lung artery walls also showed a trend towards a higher ET(A)-receptor density. The ET(B) receptor expression was low and not related to any of the above factors. Our results suggest that the paracrine lung ET-1 system is up-regulated in pediatric patients with secondary pulmonary hypertension associated with congenital heart disease. PMID- 10369114 TI - Increased serum neopterin concentrations in patients with Alzheimer's disease. AB - We measured serum neopterin concentrations in 24 patients with Alzheimer's disease (8 males, 16 females; age: 73.1+/-6.2 years; free of any infectious process) and fourteen controls of similar age (4 males, 10 females; age: 69.7+/ 8.8 years). Compared to controls, significantly higher concentrations of neopterin (p< 0.01) were found in patients with Alzheimer's disease. Among patients, concentrations of neopterin were higher in those with lower mini-mental state (p < 0.05), and an inverse correlation existed between mini-mental-state and neopterin concentrations. No such association existed with the duration of the disease. There were also significant correlations between neopterin and serum concentrations of immune activation markers such as soluble tumor necrosis factor (TNF) receptor and soluble interleukin-2 receptor (all p<0.01). Thus, increased concentrations of neopterin in serum of patients with Alzheimer's disease correlate with the severity of dementia. The data imply a chronic state of peripheral immune activation in Alzheimer's disease. PMID- 10369115 TI - Biomarkers of bone turnover and bone mineral density in hyperprolactinemic amenorrheic women. AB - We tested the hypothesis that biomarkers of bone resorption are increased in hyperprolactinemic amenorrheic patients with estrogen (E) deficiency, augmenting the possible risk of developing osteoporosis. Fifty hyperprolactinemic patients with amenorrhea of more than 12 months and with low serum E2, as well as 30 healthy fertile women (controls), matched for age and body mass index, participated in this study. Bromocriptine was administered orally to hyperprolactinemic patients and blood and urine samples were collected before and 12 weeks after treatment. Serum osteocalcin (OC) and bone-specific alkaline phosphatase (B-ALP), reflecting bone formation, and urinary deoxypridinoline (D Pyr) and N-telopeptide of type 1 collagen (NTX) excretion, reflecting bone resorption, were measured using direct immunoassays. Hyperprolactinemic patients had higher (p < 0.0005) levels of all the biomarkers compared to control values: (OC, 22+/-1.2 [SE] vs. 14+/-.99 ng/ml (+57 %); B-ALP, 14.2+/-0.7 vs. 7.5+/-0.8 ng/ml (+89 %); D-Pyr, 8.8+/-0.6 vs. 3.2+/-0.3 nmol/mmol creatinine (+175%) and NTX, 65+/-5.1 vs. 25+/-3.2 nmol bone collagen equivalent (BCE)/mmol creatinine (+160%)). These results were associated with significantly decreased lumbar spine bone mineral density (LS-BMD), measured by dual energy X-ray absorptiometry (DEXA). Treatment of hyperprolactinemia with bromocriptine restored normal values of bone formation and resorption markers. In conclusion, hyperprolactinemia with estrogen deficiency exhibits a significant increase of bone resorption which is associated with a significant decrease of LS-BMD. These changes may subject the patient to the possible risk of developing osteoporosis. PMID- 10369116 TI - Age relationships and sex differences in serum levels of pregnenolone and 17 hydroxypregnenolone in healthy subjects. AB - 17-Hydroxypregnenolone (3beta,17alpha-dihydroxypregn-5-en-20-one) and pregnenolone (3beta-hydroxypregn-5-en-20-one) were determined by radioimmunoassay following HPLC separation in serum of healthy subjects of both sexes from 2 to 66 years old (29 girls, 85 women, 30 boys, 89 men). The effects of age and sex on the levels of both steroids were investigated and the upper limits of normal in age groups were determined. The 17-hydroxypregnenolone levels as a function of age were characterized by a statistically significant maximum at the age of 18 and 20 years followed by a local minimum at the age of 39 and 37 years and by a statistically insignificant local maximum at the age of 55 and 49 years in men and women, respectively. Pregnenolone age-dependence was similar and the statistically significant maximum was reached at the age of 17 and 16 years, the local minimum occurred at the age of 37 and 38 years and the second, statistically insignificant, local maximum at the age of 48 and 47 years in men and women, respectively. Both 17-hydroxypregnenolone and pregnenolone in both sexes exhibited similarly shaped peaks with age. Both peaks of the polynomial fit in 17-hydroxypregnenolone were more pronounced in men than in women (13.0 and 9.20 nmol/l in the first peak; 7.72 and 4.78 in the second peak respectively). The situation with pregnenolone was the opposite. Both peaks of the polynomial fit in pregnenolone were lower in men than in women (2.29 and 3.21 nmol/l in the first peak; 0.92 and 1.78 in the second peak, respectively). The higher serum levels of pregnenolone at puberty and during fertile age and their wider variance in comparison with men could, be explained by the different gonadal steroidogenesis depending on the menstrual cycle, where the pregnenolone serves as a substrate for progesterone formation. The age dependencies of 17 hydroxypregnenolone and pregnenolone in women resembled that of unconjugated dehydroepiandrosterone. These results indicate that the increased metabolic activity in gonads in adolescence concerns not only dehydroepiandrosterone as the product of the 5-ene metabolic pathway but also its precursors. PMID- 10369117 TI - A study of hypermagnesaemia in a hospital population. AB - The purpose of this present study was to assess the prevalence of hypermagnesaemia in a hospital population. Furthermore, the relationship between hypermagnesaemia and other common electrolyte disturbances such as hypo- and hypercalcaemia, hypo- and hyperkalaemia and hypo- and hyperphosphataemia was studied. Twenty-seven percent of magnesium requests showed a serum magnesium concentration equal to, or greater than, 1.0 mmol/l. Hyperkalaemia (a plasma potassium concentration of equal to, or greater than, 5.0 mmol/l) was found in 18% of the patients with hypermagnesaemia and 25 % of these patients showed hyperphosphataemia (a plasma phosphate concentration of equal to, or greater than, 1.5 mmol/l). Of the serum magnesium requests, hypermagnesaemia was particularly common on the intensive care (23%) and the renal unit (43%). Hypermagnesaemia was also seen in patients undergoing cardiothoracic surgery (17 %) and who had an acute myocardial infarction (8 %). Seventy-three percent of patients with a plasma magnesium of greater than 1.0 mmol/l showed abnormal renal function. However, it was rare to find a serum magnesium of greater than 2.0 mmol/l (less than 1% of magnesium requests). PMID- 10369118 TI - Multiple regression analysis of interference effects from a hemoglobin-based oxygen carrier solution. AB - The use of hemoglobin-based oxygen carrier solutions in patients requiring blood transfusion will necessitate that clinical laboratories have mechanisms in place to evaluate the potential interference effect of these substances on testing methods. Because these oxygen carrier solutions contain acellular hemoglobin, but do not contain many of the intracellular enzymes and ions present in erythrocytes, interference effects from blood substitutes may be quite different when compared to in vivo or in vitro lysis of erythrocytes. We evaluated the potential interference effect of Diaspirin Cross-linked Hemoglobin on 29 different clinical laboratory analytes. Various combinations of these analytes were tested using the Hitachi 747 and 911 systems, a Beckman CX3, an Abbott AxSym, a Bayer Immuno I, and a Dade ACA IV; a total of 60 analyte/instrument combinations. We used the method of multiple regression analysis to classify interferences as analyte-dependent, analyte-independent, or a combination of the first two types. The presence of clinically significant test interference was derived by using the criteria for maximum allowable error specified in the Clinical Laboratory Improvement Amendments of 1988. Using these criteria, we found significant interference from Diaspirin Cross-linked Hemoglobin with 13 of 29 analytes tested. Interference was noted with the Hitachi 747 and 911 methods for albumin, alkaline phosphatase, total and conjugated bilirubin, cholesterol, total carbon dioxide, iron, lactate dehydrogenase, magnesium, total protein, and triglyceride. In addition, Diaspirin Cross-linked Hemoglobin interfered with measurement of L-lactate using the ACA IV and minor interference was noted with glucose measured using the Beckman CX3. Data from the interference studies was graphically displayed in the form of interference plots. These plots show the maximum allowable test error, due to Diaspirin Cross-linked Hemoglobin, as a function of analyte and interferent concentrations. Evaluation of the potential interference effect of hemoglobin-based oxygen carrier solutions with use of multiple regression analysis and graphical display of the resultant data in the form of interference plots allows for more reliable reporting of test results from specimens containing these products. PMID- 10369120 TI - Automated prozone effect detection in ferritin homogeneous immunoassays using neural network classifiers. AB - The application of turbidimetric homogeneous immunoassays made the determination of several plasma components widely available. The sensitivity and accuracy of these assays are appropriate enough for routine laboratory use; however, in the case of many pathologically high concentration samples, prozone effect (high dose hook effect) can be observed, that leads to false-negative determination. Up to the present there are no cost-effective algorithms available for the safe detection of the prozone effect. Pathological serum ferritin values can be elevated up to 5000 ng/ml, while the measuring range covers only the 0-440 ng/ml range by a commercial assay. The determination of samples with ferritin concentration higher than 1500 ng/ml results in false-negative values because of the overlapping measuring range and prozone effect range. The prozone effect can be recognised by analysis of reaction kinetics after measurement. We have developed a neural network classifier system to analyse reaction kinetics of the measurements and check the prozone effect. One thousand five hundred determinations and 77 patient samples were used for neural network training and test. Using the trained neural networks, false-negative results can be filtered immediately after the determination, without re-run; thus, the sensitivity of plasma ferritin determination may become reliable enough, even in the case of high concentration samples. Applying this new technology, false-negative serum ferritin determinations can be avoided, thus even a relatively high hook effect rate (5-12% in different patient groups) can be handled safely. PMID- 10369119 TI - Serum ionized magnesium: comparison of results obtained with three ion-selective analyzers. AB - In a two-center (Academic Medical Center, The Netherlands, and National Institutes of Health, USA) study, we compared ionized magnesium (iMg2+) results in serum determined with the AVL 988/4, KONE Microlyte 6 and NOVA CRT, which are the currently available analyzers equipped with a magnesium ion-selective electrode. The comparison was performed with frozen serum samples from normal individuals and patients. Imprecision and reference intervals were established. We found the best agreement between the KONE(x) and AVL(y) magnesium ion selective electrodes (y= 0.972x-0.013; n=138) with samples from patients. With samples from normals, all three analyzers reported significantly different results (p<0.05). Best precision was found using the NOVA; coefficients of variation established at three levels were all < 4.0%. Coefficients of variation for the AVL and KONE were <5% at normal and high iMg2+, but 10.7 and 9.4%, respectively, at iMg2+ approximately 0.30 mmol/l. The reference intervals (mean+/ standard deviation) based on measurements in fresh serum samples were different for each analyzer: 0.55-0.63 mmol/l for AVL, 0.470.57 mmol/l for KONE and 0.43 0.55 mmol/l for NOVA. Thus, significant differences among the ionized magnesium concentration obtained with the three analyzers, limit comparison of results in clinical practice, and need to be resolved (e.g. by improvement of specificity and standardization of calibrators). PMID- 10369121 TI - Recent activities of EC4 in the harmonization of clinical chemistry in the European Union. AB - This article describes the recent activities of the European Communities Confederation of Clinical Chemistry (EC4). Main goal of EC4 is harmonization of clinical chemistry in the European Union and Europe. EC4's actions connected to that are training and registration of professionals, and accreditation of laboratories. The 35000 professionals practising clinical chemistry in the EU have different backgrounds (medical, pharmaceutical, science-oriented, veterinary, or microbiological). Thus, for the harmonization of training of clinical chemists, EC4 has published a European Syllabus for Postgraduate Training, and instituted a European Union Register for Clinical Chemists. The Syllabus is an indication of the level of requirements in postgraduate training. The EC4 initiative to implement the European Register for Clinical Chemists is based on the 8 years vocational training necessary to obtain sufficient knowledge in clinical chemistry according to the European Syllabus. A guide to the EC4 Register has been published; registration leads to the title European Clinical Chemist (EurClinChem). The accreditation of laboratories must be based on a total quality management system. EC4 has described guidelines (essential criteria) which it judges appropriate for establishing the quality of medical laboratory service; it does not wish to fulfil the role of an accrediting body. Moreover, a working group has been set up to seek to harmonize the work of national accrediting bodies. Therefore, it is logical that EC4 monitors the activities of the different standardizing bodies that might influence the practice of clinical chemistry in the EU. Finally, some aspects concerning the future strategy of EC4 are brought forward. PMID- 10369122 TI - Protein A of Staphylococcus aureus evokes Th1 type response in mice. AB - Protein A (PA) of Staphylococcus aureus is known to elicit several cytokines such as IFN gamma, TNF alpha and IL1. However, it has not been delineated yet as to which differentiation pathway lymphocytes follow after stimulation by PA. In this report, we attempted to collect such evidences. Cytokines, such as IFN gamma, IL2, IL4, IL6, IL10, TNF alpha, IL1alpha and IL1beta were measured in serum by ELISA. Our results show that 1 microg dose of PA stimulates the production of IFN gamma (115 +/- 5 pg/ml), TNF alpha (250 +/- 8 pg/ml) and IL1alpha (100 +/- 5 pg/ml) as compared to control levels of, 22 +/- 2, 20 +/- 2 and 35 +/- 3 pg/ml respectively whereas IL2 and IL1beta secretion were less (beyond the lower detection limit of the kit and 25 +/- 1 pg/ml, respectively) as compared to control (28 +/- 2 and 52 +/- 4 pg/ml, respectively). Larger dose of PA (10 microg) increases the expression of IL2 (75 +/- 3 pg/ml), TNF alpha (1380 +/- 120 pg/ml), IL1alpha (495 +/- 10 pg/ml) and IL1beta (110 +/- 7 pg/ml) as compared to controls described above. We also observed that 1 microg dose of PA decreases IL4, IL6 and IL10 secretion to 9 +/- 1, 10 +/- 1 and 10 +/- 2 pg/ml, respectively, whereas 10 microg dose also decreased them to 11 +/- 1, 12 +/- 2 and 30 +/- 4 pg/ml, respectively as compared to the background controls, i.e. 50 +/- 5, 50 +/- 2 and 215 +/- 9 pg/ml respectively. The ratio of IFN gamma to IL4 increased and the peak value at 4 h, came to 13 +/- 1 and 9.6 +/- 0.5 with 1 microg and 10 microg PA, respectively, which is an established parameter indicating a Th1 type response. Flow cytometry analysis of CD4+/CD8+ cells, and c myc protein expression by splenocytes indicate that 1 microg dose of PA causes 2 fold increase of CD4+ cells with no change in CD8+ cells, and 10-fold increase in c-myc protein, whereas 10 microg dose increases CD4+ cells 4-fold, CD8+ cells 3 fold and c-myc protein 100-fold. The cell cycle data shows an induction of apoptosis in thymocytes and splenocytes with the large dose (10 microg), whereas the 1 microg dose does not show any apoptosis. This report indicates that a Th1 response is induced in mice, after PA inoculation at a dose of 1 microg animal. Thus, cytokine mediated therapeutic strategies should consider the fact that an induction of large concentration of some cytokines might become detrimental to the host. PMID- 10369123 TI - Identification of peptides presented by HLA class I molecules on cervical cancer cells with HPV-18 infection. AB - In this work we eluted peptides from purified class I MHC molecules, isolated from a novel human cervical carcinoma cell line (INBL), generated in our laboratory and positive for HPV-18 infection. A fraction of these peptides was capable of stimulating T lymphocytes obtained from a donor matched for HLA-Cw4 and who was also HPV-18+. Direct N-terminal Edman degradation of these peptides, revealed the sequence (XQFPIFLQF) that matched 85% with the sequence NVFPIFLQM localized in between the 54 and 62 residues of the HPV-18 L1 protein. After stimulation with the synthetic peptide NVFPIFLQM, T lymphocytes from the donor were capable to lyse INBL cells. Our results provide evidence of the existence of naturally occurring viral epitopes presented on cervical cancer cells by the HLA Cw4 allele, that could be useful for immunotherapy on this type of patient. PMID- 10369124 TI - Flow cytometric analysis of the molecular mechanisms of immunosuppressive action of the active metabolite of leflunomide and its malononitrilamide analogues in a novel whole blood assay. AB - Malononitrilamides (MNAs) are a new class of immunomodulatory drug highly effective in in vivo models of allo- and xenotransplantation. Knowledge of their effects on immune cells, however, is limited and has been derived solely from investigations using isolated mononuclear cells. This use of purified cells to investigate drug activity is not ideal, so we have combined the analytical power of flow cytometry with our mitogen-driven, whole blood lymphocyte activation and proliferation assays to investigate the in vitro mechanism of action of MNAs. We first show that MNAs (A77 1726, HMR1279, and HMR1715), as well as brequinar (BQR) and cyclosporine (CsA), effectively inhibit cell activation antigen expression and lymphocyte proliferation. We next show that the inhibitory effects of MNAs and BQR, but not CsA, are reversed by the addition of uridine to the culture. These results suggest that inhibition of pyrimidine biosynthesis may be a mechanism by which MNAs suppress both lymphocyte activation and proliferation since these effects were reversed when uridine nucleotide pools were replenished. This novel finding of suppression of activation antigen expression by MNAs in whole blood expands our understanding of the effects of this new class of drug. PMID- 10369125 TI - Dual specificity of a human neutralizing monoclonal antibody, specific for the V3 loop of GP120 (HIV-1). AB - Neutralizing antibodies specific for the third variable (V3) domain of gp120, the HIV-1 surface envelope protein, appear early in infection. However, they are usually highly specific for the priming isolate. To identify potential mimotopes of the V3 domain, we have screened a hexapeptide phage library with a human neutralizing mAb, mAb 268, specific for the V3 loop of the viral MN isolate. We have identified two groups of sequences. Within the first group, sequence 268-1 reproduces the linear epitope identified using a conventional epitope mapping approach. The sequence 268-1, H L G P G R, corresponds to amino acids 315-320, localized in the highly conserved tip of the V3 loop. A second group of sequences was identified, including sequence 268-2, K A I H R I. Partial homology with a more variable region of the V3 loop can be found. Using synthetic peptides, we demonstrated that peptides, 268-1 and 268-2, both interact with the same binding site as the V3 region on the 268 mAb. Moreover, both peptides can inhibit the interaction of the 268 mAb with the original immunogen, gp120MN. Peptide 268-1 can compete with peptide 268-2, albeit poorly, for binding of the 268 mAb. When injected into rabbits, KLH conjugated peptide 268-2 elicited antibodies that interact specifically with the initial immunogen gp120MN. These data suggest that peptide 268-2 is both an antigenic and immunogenic mimic of the natural antigen, gp120MN. PMID- 10369126 TI - Galectin-1, a natural ligand for the receptor-type protein tyrosine phosphatase CD45. AB - Galectin-1 binds preferentially to N-acetyllactosamine residues on oligosaccharides associated with several cell surface glycoconjugates. In the present work, placental galectin-1 has been identified to be a natural ligand for the receptor-type protein tyrosine phosphatase CD45. The binding of galectin-1 to CD45 was detected by affinity chromatography of NP 40 solubilized Jurkat T cell membranes on galectin-1 agarose followed by immunoblotting of the galectin-1 agarose bound fraction applying monoclonal antibodies to CD45 isoforms. The PTPase activity of the galectin-1 agarose binding membrane fraction could be inhibited by sodium orthovanadate. Preincubation of Jurkat T cell membrane preparations with galectin-1 decreased the membrane-associated PTPase activity in a concentration-dependent manner. Incubation of Jurkat cells with galectin-1 suppressed the immunoprecipitated PTPase activity of CD45. Galectin-1 stimulates the cell surface expression of phosphatidylserine an early indicator of apoptosis. In CD45+ Jurkat T cells, galectin-1 induces higher levels of phosphatidylserine when compared with CD45- Jurkat cells. These observations indicate that galectin-1-mediated ligation of CD45 is involved in the induction of apoptosis in Jurkat T cells. PMID- 10369127 TI - Linomide downregulates autoimmunity through induction of TH2 cytokine production by lymphocytes. AB - Linomide is a synthetic immunomodulator that has been shown to protect animals against a wide range of spontaneously developing or induced autoimmune diseases. We have previously reported that Linomide blocks both the clinical and the histopathological manifestations of experimental autoimmune encephalomyelitis (EAE) in various animal models. In this study, in an effort to elucidate the mechanisms by which Linomide suppresses EAE, and autoimmunity in general, we investigated the in vivo effects of this drug on the TH1/TH2 lymphocyte balance, which is important for the induction or inhibition of autoireactivity. Naive SJL/J mice were treated orally for 15 days with Linomide (80 mg/kg/day). Spleen cells were obtained at various time points during the treatment period and were stimulated in vitro with concanavalin A. Interleukins IL-4, IL-10 and IL-12, transforming growth factor-beta (TGFbeta) and interferon-gamma (IFNgamma) cytokine production was evaluated both by means of detection of the cytokines in the medium (by ELISA technique) and by detection of the cytokine mRNA production, using a semiquantitative reverse transcriptase polymerase chain reaction method. A significant upregulation of IL-4, IL-10 and TGFbeta was observed following treatment with Linomide, which peaked at day 10 (IL-10) or day 15 (IL-4). On the other hand, IL-12 and IFNgamma production were either unchanged or decreased. It seems therefore that Linomide induces in vivo a shift towards TH2 lymphocytes which may be one of the mechanisms of downregulation of the autoimmune reactivity in EAE. Our observations indicate that downregulation of TH1 cytokines (especially IL-12) and enhancement of TH2 cytokine production may play an important role in the control of T-cell-mediated autoimmunity. These data may contribute to the design of new immunomodulating treatments for a group of autoimmune diseases. PMID- 10369128 TI - Heat-labile enterotoxin of Escherichia coli and its site-directed mutant LTK63 enhance the proliferative and cytotoxic T-cell responses to intranasally co immunized synthetic peptides. AB - The adjuvanticity of heat-labile enterotoxin (LT) of Escherichia coli and its non toxic mutant LTK63 was assessed and compared for intranasal immunization of synthetic peptides. Mice immunized intranasally with LT, or its mutant LTK63, generated strong systemic proliferative and cytotoxic T-cell responses to co administered synthetic peptides. The wild LT toxin promoted higher peptide specific proliferative and cytotoxic T-cell responses than the LTK63 mutant. Moreover, the wild-type LT toxin was shown to promote peptide-specific memory CTL responses which were detectable 1 year after intranasal priming. Both LT and LTK63 molecules were shown to be immunogenic, with serum antibody subclasses being predominantly IgG1 and to a lesser extent IgG2a. These findings demonstrate that cellular immune responses to small synthetic peptide antigens administered by the intranasal route can be potentiated with the use of mucosal adjuvants. Moreover, the ability of LT and LTK63 to promote both CD4+ and CD8+ T-cell responses will have relevance to the design and production of future mucosal vaccines. PMID- 10369130 TI - Thrombin-induced interleukin-8 production and its regulation by interferon-gamma and prostaglandin E2 in human monocytic U937 cells. AB - Blood coagulation and inflammation pathways are linked in many aspects. A number of serum factors involved in coagulation cascades affect directly or indirectly inflammatory responses, whereas proinflammatory cytokines influence blood coagulation pathways. In this work we demonstrated that thrombin is an effective stimulus in inducing interleukin (IL)-8 expression in human monocytic cell line U937. IL-8 induction was found at the mRNA and protein levels. The effect of thrombin on IL-8 production was mimicked by thrombin receptor-activating peptide indicating that thrombin effect was mediated by the specific receptor for thrombin. Moreover, thrombin-induced IL-8 production by U937 cells was differentially regulated by interferon (IFN)-gamma and prostaglandin (PG)E2. While IFN-gamma enhanced thrombin-induced IL-8 production, PGE2 acted as a negative regulator. Taken together, thrombin may play an important role in communication between blood coagulation and inflammation by inducing IL-8 production by monocytes and this role for thrombin may be further regulated by lymphokines and lipid mediators. PMID- 10369129 TI - Immune responses to P. falciparum-MSP1 antigen: lack of correlation between antibody responses and the capacity of peripheral cellular immune effectors to respond to this antigen in vitro. AB - Protective immunity to P. falciparum blood stage infection is thought to be dependent on IgG antibodies, although the mechanisms that underlie such immunity are not clearly understood. One of the antigens thought to be involved in this protective response is MSP1. The present study has examined the levels and distribution of IgG (and IgM) antibodies to the C-terminal 19 kDa fragment of MSP1 in plasma from P. falciparum immune adult Senegalese and the capacity of the peripheral blood mononuclear cells from these patients to either proliferate or secrete IFN-gamma, IL-10 or IL-4 in vitro in response to this antigen. Specific antibodies were found in 74% of individuals' plasma; 44% of mononuclear cells proved capable of proliferating in vitro and IFN-gamma, IL-10 and IL-4 were detected in 37, 23 and 0% of culture supernatants, respectively. No significant association was found between the presence of antibodies and immune cell reactivity under the culture conditions used. This study emphasizes the complexity of the mechanisms responsible for the sustained production of potentially protective antibodies in response to proposed T-cell dependent P. falciparum blood stage antigens. PMID- 10369131 TI - Secretion of stem cell factor by alveolar fibroblasts in interstitial lung diseases. AB - Sarcoidosis (SA) and diffuse interstitial fibrosis (DIF) are characterized by alveolitis, mast cell hyperplasia and increased fibroblast proliferation. Stem cell factor (SCF) stimulates proliferation of hematopoietic progenitor cells involved in mast and stromal cell interaction. We assessed the role of SCF secreted by alveolar fibroblasts (AFb) in the development of fibrosis of DIF and SA in six patients with SA and six patients with DIF. Bronchoalveolar lavage (BAL) was performed by conventional methods. A total of 500 cells were differentially counted from Giemsa-stained cytopreps. AFb and supernatants were recovered from long-term cultures of BAL cells and from 24 h cultures of confluent AFb. Levels of SCF were measured by ELISA. Alpha actin content of AFb was characterized by immunohistochemistry. The expression of AFb mRNA for IL1 alpha and beta, TGF-beta, IFN-gamma, IL-2, IL-4, IL-5 and IL-6 was determined by RT-PCR. There was a lymphocytic predominance in the SA patients and an increase in neutrophils and eosinophils in DIF. SCF secreted by AFb from DIF was significantly higher than in SA. TNF + IL-1 significantly decreased the secretion of SCF by AFb. There was a positive correlation between SCF levels and the percentage eosinophils but not for metachromatic cells. Alpha-actin expression of AFb in DIF was significantly higher than in SA. Cytokine mRNA was extracted from AFb of two SA and two DIF patients. The profile showed that only in stimulated AFb isolated from the DIF patients can IL-5 transcripts be visualized. In conclusion, AFb can contribute to the onset of fibrosis by secreting SCF and IL-5 which, in turn, may recruit eosinophils. PMID- 10369132 TI - Inhibition of PHA-induced cell proliferation by polyclonal CD4 antibodies generated by DNA immunization. AB - Although the role of CD4 molecule as associative binding element to MHC class II is well documented, their role in T cell activation is unclear. In the present report we used DNA immunization, which is currently shown to induce potent immune responses, to produce the polyclonal antibodies specific for the CD4 molecule and used the generated antibodies to characterize the CD4 function. A rabbit was pre treated with bupivacaine hydrochloride for 24 h which was followed by intramuscular injection of DNA encoding CD4 protein (CD4-DNA) at weekly interval. By this procedure, CD4 antibodies were detected in the immunized serum after two DNA inoculations. The CD4 antibodies titer was up to 1:800 after five DNA inoculations. The rabbit polyclonal CD4 antibodies recognized both recombinant CD4 protein expressed on CD4-DNA transfected COS cells and native CD4 protein presented on peripheral lymphocytes and CD4+ cell lines. These generated CD4 antibodies could block the binding of standard CD4 mAb, Leu3a and 13B8.2, to the CD4 molecule. To characterize the function of CD4 molecule, PBMC were cultured in the presence of sub-optimal dose of PHA and the produced polyclonal CD4 antibodies. We found that the polyclonal CD4 antibodies strongly suppressed PHA induced cell proliferation. The inhibitory effect of CD4 antibodies may be due to their steric inhibition of the CD4-TCR/CD3 association or may interfere with the binding of CD4 to its ligand IL-16, resulting in the reduction of signal transduction and subsequent cellular responses. Our results indicate the possibility of utilizing DNA immunization to produce polyclonal antibodies against cell surface molecule. PMID- 10369133 TI - Generation and function of bone marrow-derived dendritic cells from CD4/CD8(-/-) double-knockout mice. AB - We present a novel, simple and straightforward method to obtain mouse bone marrow derived dendritic cells (DC) from C57Bl/6 CD4/CD8(-/-) double knock-out mice. This new method, involving culture of bone marrow cells in medium supplemented with Interleukin 4 and Granulocyte-Macrophage Colony-Stimulating Factor, does not involve negative immunodepletion of CD4+ and CD8+ populations, or extensive prior manipulations of the starting population. The resulting, loosely adherent cell population, exhibited the morphological characteristics and typical surface markers of DCs, and were endowed with the functional activities characteristic of bone marrow-derived DCs of wild-type mice. Interestingly, LCMV GP33-41 peptide loaded CD4/CD8(-/-) DCs were efficiently lysed by peptide-specific activated CTLs in vitro. Furthermore, these peptide-loaded CD4/CD8(-/-) DCs induced a peptide specific CTL response upon immunization of wild-type C57Bl/6 mice. PMID- 10369134 TI - Low affinity Fc gamma receptors on murine macrophages: mitogen-activated protein kinase activation and AP-1 DNA binding activity. AB - Mouse macrophage cell lines such as J774 express Fc receptors for IgG2a immune complexes, which upon binding of the proper ligand, trigger several signal transduction pathways. A surface to nucleus signaling through these receptors has been demonstrated. We describe here the activation of the mitogen-activated protein kinase (MAPK) and an increase in the binding of the activator protein 1 (AP-1) to DNA upon receptor stimulation. The described effects are only partially blocked by inhibitors of the Ca2+/diacylglycerol-dependent protein kinase (PKC), suggesting that differential signaling pathways are activated upon receptor cross linking and that they converge at or above the MAPK level. These results pave the way to our understanding of Fc gammaR cross-linking induced gene expression regulation. PMID- 10369135 TI - CD8+ lymphocyte-mediated suppression of human immunodeficiency virus type 1 expression in human brain cells. PMID- 10369136 TI - Production of pro-inflammatory cytokines in human monocytes: not a cascade but the dependence on protein kinase C pathway. PMID- 10369137 TI - Basal ganglia and gait control: apomorphine administration and internal pallidum stimulation in Parkinson's disease. AB - Gait coordination was analyzed (four-camera 100 Hz ELITE system) in two groups of idiopathic Parkinson disease (PD) patients. Five patients underwent continuous infusion of apomorphine and were recorded in two different sessions (APO OFF and APO ON) in the same day. Three patients with a previous chronic electrode implantation in both internal globi pallidi (GPi) were recorded in the same experimental session with the electrodes on and off (STIM ON and STIM OFF). The orientation of both the trunk and the lower-limb segments was described with respect to the vertical in the sagittal plane. Lower-limb inter-segmental coordination was evaluated by analyzing the co-variation between thigh, shank, and foot elevation angles by means of orthogonal planar regression. At least 30 gait cycles per experimental condition were processed. We found that the trunk was bent forward in STIM OFF, whereas it was better aligned with the vertical in STIM ON in both PD groups. The legs never fully extended during the gait cycle in STIM OFF, whereas they extended before heel strike in STIM ON. The multisegmental coordination of the lower limb changed almost in parallel with the changes in trunk orientation. In STIM OFF, both the shape and the spatial orientation of the planar gait loops (thigh angle vs. shank angle vs. foot angle) differed from those of physiological locomotion, whereas in STIM ON the gait loop tended to resume features closer to the control. Switching the electrodes on and off in patients with GPi electrodes resulted in quasi-parallel changes of the trunk inclination and of the planar gait loop. The bulk of the data suggest that the basal-ganglia circuitry may be relevant in locomotion by providing an appropriate spatio-temporal framework for the control of posture and movement in a gravity based body-centered frame of reference. Pallido-thalamic and/or pallido mesencephalic pathways may influence the timing of the inter-segmental coordination for gait. PMID- 10369138 TI - Cortical control of optokinetic nystagmus in humans: a positron emission tomography study. AB - Positron emission tomography (PET) was used to address the issue of physiological changes in the cerebral cortex associated to optokinetic nystagmus (OKN) in humans. We studied regional cerebral blood flow in eight volunteers during reflexive induction of OKN by a pattern of dots moving unidirectionally (toward the left side). We used two control conditions, with subjects passively viewing either stationary or incoherently moving dots. This paradigm was designed in order to differentiate the OKN-related activations from blood flow changes related to visual motion. When compared with the stationary condition, OKN activated a set of occipital areas known to be sensitive to visual motion. Bilateral activation was found in the striate cortex (V1) and the parieto occipital fissure, while area V5, the intraparietal sulcus, and the pulvinar were activated only in the left hemisphere. When compared with incoherent motion, OKN activated the V1 and the parieto-occipital fissure bilaterally and the right lingual gyrus, while a signal decrease was observed in the V5 region in both hemispheres. No significant signal changes were found in areas implicated in saccades or in processing vestibular information. These results indicate that processing of OKN-related information is associated with neural activity in a specific set of visual motion areas and suggest that this network can be asymmetrically activated by a strictly unidirectional stimulation. Results are also discussed in terms of the specific kinds of OKN-related information processing subserved by each area in this network. PMID- 10369139 TI - Thalamocortical inputs trigger a propagating envelope of gamma-band activity in auditory cortex in vitro. AB - To investigate how auditory cortex responds to thalamic inputs, we have used electrophysiological and anatomical techniques to characterize a brain slice containing functionally linked thalamocortical and intracortical pathways. In extracellular recordings, stimulation of thalamic afferents elicited a short latency field potential and current sink in layer IV of the cortex, followed by 100-500 ms of polysynaptic activity containing rapid (gamma-band, 20-80 Hz) fluctuations. Paired intracellular and extracellular recordings showed that a short-latency excitatory postsynaptic potential (EPSP) corresponded to the fast extracellular potential, and that a slow intracellular depolarization with superimposed rapid fluctuations corresponded to the polysynaptic extracellular activity. Pharmacological manipulations demonstrated that glutamate receptors contributed to mono- and polysynaptic activity, and that the gamma-band fluctuations contained intermixed rapid depolarizations and Cl(-)-mediated inhibition. The spread of evoked activity through auditory cortex was determined by extracellular mapping away from the excitatory focus (the site of the largest amplitude fast response). The short-latency potential traversed auditory cortex at 1.25 m/s and decreased over 1-2 mm, likely reflecting sequential activation of cells contacted by thalamocortical arbors. In contrast, polysynaptic activity did not decrease but propagated as a spatially restricted wave at a 57-fold slower velocity (0.022 m/s). Thus, stimulation of the auditory thalamocortical pathway in vitro elicited a fast glutamatergic potential in layer IV, followed by polysynaptic activity, including gamma-band fluctuations, that propagated through the cortex. Propagating activity may form transient neural assemblies that contribute to auditory information processing. PMID- 10369140 TI - Listing's plane rotation with convergence: role of disparity, accommodation, and depth perception. AB - Earlier studies have reported temporal rotation of Listing's plane with convergence of the eyes causing torsion, which is dependent on eye elevation. The amount by which the planes rotate differs from study to study. To gain insight into the functional significance of the temporal tilt of Listing's plane for vision, we examined whether the rotation of the plane depends on the visual conditions, namely on the stimuli driving vergence. In different conditions, accommodative vergence, disparity-vergence, combinations of disparity with accommodation or depth perception were used and the resulting rotation of Listing's plane was measured. Our findings show, for the first time, that the relationship between convergence and Listing's-plane temporal rotation depends on the stimuli driving vergence. When the stimulus contains only disparity cues, vergence and Listing's plane rotate immediately and consistently among subjects. Accommodative vergence, the mutual couplings between vergence and accommodation, can influence the orientation of Listing's plane, but they do so in a idiosyncratic way. The largest rotation was elicited by stereograms combining disparity-vergence with depth perception. These findings support the idea of a functional role of Listing's plane rotation for binocular vision, perhaps for depth perception. PMID- 10369141 TI - Three-dimensional extraocular motoneuron innervation in the rhesus monkey. I: Muscle rotation axes and on-directions during fixation. AB - The rotation axis for each of the six extraocular muscles was determined in four eyes from three perfused rhesus monkeys. Measurements of the locations of muscle insertions and origins were made in the stereotaxic reference frame with the x-y plane horizontal and the x-z plane sagittal. The computed rotation axes of the horizontal recti were close to being in the x-z plane at an angle of about 15 degrees to the z axis. The rotation axes of the vertical recti and the obliques were close to being in the x-y plane at an angle of about 30 degrees to the y axis. In five alert rhesus monkeys, we simultaneously recorded extraocular motoneuron activity and eye position in three dimensions (3D). The activity of 51 motoneuron axons was obtained from the oculomotor (n=34), trochlear (n=11), and abducens nerve (n=6) during spontaneous eye movements. To extend the torsional range of eye position, the animals were also put in different static roll positions, which induced ocular counterroll without dynamic vestibular stimulation. Periods of 100 ms during fixation or slow eye movements (<10 degrees/s) were chosen for analysis. For each motoneuron, a multiple linear regression was performed between firing frequency and 3D eye position, expressed as a rotation vector, in both stereotaxic and Listing's reference frame. The direction with the highest correlation coefficient (average R=0.94+/-0.07 SD) was taken as the on-direction. Each unit's activity could be unequivocally attributed to one particular muscle. On-directions for each motoneuron were confined to a well-defined cone in 3D. Average on-directions of motoneurons differed significantly from the corresponding anatomically determined muscle rotation axes expressed in the stereotaxic reference frame (range of deviations: 11.9 degrees to 29.0 degrees). This difference was most pronounced for the vertical recti and oblique muscles. The muscle rotation axes of the vertical rectus pair and the oblique muscle pair form an angle of 58.3 degrees, whereas the corresponding angle for paired motoneuron on-directions was 105.6 degrees. On-directions of motoneurons were better aligned with the on-directions of semicircular canal afferents (range of deviation: 9.4-18.9 degrees) or with the anatomically determined sensitivity vectors of the semicircular canals (range of deviation: 3.9-15.9 degrees) than with the anatomically determined muscle rotation axes, but significant differences remain to be explained. The on-directions of motoneurons were arranged symmetrically to Listing's plane, in the sense that the torsional components for antagonistically paired muscles were almost equal, but of opposite sign. Thus, the torsional components of motoneuron on-directions cancel when eye movements are confined to Listing's plane. This arrangement simplifies the neuronal transformations for conjugate head-fixed voluntary eye movements, while the approximate alignment with the semicircular canal reference frame is optimal for generating compensatory eye movements. PMID- 10369142 TI - Development of anticipatory postural adjustments in a bimanual load-lifting task in children. AB - Anticipatory postural adjustments (APA) are needed to perform a movement without perturbing posture. We investigated the development of APA in 3- to 4-year-old children during a bimanual load-lifting task. The task required maintaining a stable elbow position despite imposed or voluntary unloading of the forearm. Although children can compensate the consequences of unloading by using APA, their performance did not reach an adults' level. In addition, children showed high intra-individual variability in the voluntary situation, revealed by the coexistence of both adult-like and immature patterns in kinematic and electromyographic data. In conclusion, the present study reports that APA, associated with a bimanual load-lifting task, are still being set up in 3- to 4 year-old-children. The intra-individual variability should decrease with age and be associated with a progressive mastering of the timing parameters characterizing APA. PMID- 10369143 TI - Grip forces exerted against stationary held objects during gravity changes. AB - In the present study, grip forces exerted against a stationary held object were recorded during parabolic flights. Such flight maneuvers induce changes of gravity with two periods of hypergravity, associated with a doubling of normal terrestrial gravity, and a 20 s period of microgravity. Accordingly, the object's weight changed from being twice as heavy as normally experienced and weightless. Grip-force recordings demonstrated that force control was seriously disturbed only during the first experience of hyper- and microgravity, with the grip forces being exceedingly high and yielding irregular fluctuations. Thereafter, however, grip force traces were smooth, the force level was scaled to the object's weight under normal and high-G conditions, and the grip force changed in parallel with the weight during the transitions between hyper- and microgravity. In addition, during weightlessness, when virtually no force was necessary to stabilize the object, a low force was established, which obviously represented a reasonable safety margin for preventing possible perturbations. Thus, all relevant aspects of grip-force control observed under normal gravity conditions were preserved during gravity changes induced by parabolic flights. Hence, grip-force control mechanisms were able to cope with hyper- and microgravity, either by incorporating relevant receptor signals, such as those originating from cutaneous mechanoreceptors, or by adequately including perceived gravity signals into control programs. However, the adaptation to the uncommon gravity conditions was not complete following the first experience; finer tuning of the control system to both hyper- and microgravity continued over the measurement interval, presumably with a longer observation period being necessary before a stable performance can be reached. PMID- 10369144 TI - Importance of attentional mechanisms in audiovisual links. AB - The effect of auditory cues at different levels of visual processing was examined by using a visual "conjunction of features" discrimination task (experiment 1) and a "feature" discrimination task (experiment 2). In both experiments the visual target, appearing either on the left or the right of Ss' midline, was preceded by a brief tone either spatially proximal or distal to the target. In the "conjunction" task, subjects had to discriminate the orientation of a T flanked with T distractors of different orientations. In this task, assumed to require focused attention, discrimination accuracy was increased when the sound cue occurred at the subsequent visual target location and was decreased when it occurred at its alternative location. In the "feature" experiment, subjects had to discriminate the orientation of a line segment (+/-45 degrees) presented among line segment distractors. Accuracy was not affected, either when the sound was proximal or when it was distal to the location of the visual target. Results suggest that the early processing of sensory information is modality specific and that interference of auditory stimulation with visual stimuli is more pronounced as the processing of visual stimuli requires focused attention. PMID- 10369145 TI - Motor coordination and spatial orientation are affected by neurofilament maldistribution: correlations with regional brain activity of cytochrome oxidase. AB - NFH-LacZ transgenic mice are characterized by an early accumulation of the neurofilament cytoskeleton in the cell bodies of neurons with age-associated abnormalities of motor neurons and cerebellar Purkinje cells. In comparison to normal littermate controls, irrespective of age (3 and 12-20 months), NFH-LacZ transgenic mice had a lower number of rears in an open field, deficiencies in some motor-coordination tests, and a higher number of quadrant entries and escape latencies while swimming toward a visible platform. Decreased cytochrome oxidase activity in the lateral reticular nucleus of NFH-LacZ mice was associated with poor performance in two motor coordination tests. Lower metabolic activity in the lateral reticular nucleus may be secondary to previously described cerebellar abnormalities, leading to deficient motor control. The dramatic cytoskeletal perturbation characterizing NFH-LacZ mice affects only selective neuronal populations and results in selective behavioral deficits, which can be correlated with regional brain metabolic activity. PMID- 10369146 TI - Inhibitory control of competing motor memories. AB - The ability to inhibit previously learned visuomotor associations is essential for efficient learning of novel behaviors. While the neural basis of the system that might control interactions between competing motor memories is not known, it has been demonstrated that animals with ventral and orbital prefrontal cortex (PFC) deficits have particular difficulties in learning to withhold responses to previously conditioned sensory stimuli. Here we measured regional cerebral blood flow (rCBF), using positron emission tomography, during learning of a novel motor task that required inhibition of a previously learned motor memory. Subjects (n=24) learned reaching movements in a force field (field A). After a variable time interval, some subjects (n=15) learned to reach in a field with a reversed pattern of forces (field B). When the time interval was short (10 min), learning in field B was coincident with a reactivation of regions that had become initially activated during learning in field A: the left putamen and bilaterally in the dorsolateral PFC. Behaviorally, this was accompanied with perseveration that lasted for hundreds of movements, suggesting an instantiation of the internal model for field A during learning in field B. Neither the reactivation nor the perseveration were observed in a different group of subjects that learned field B at 5.5 h. We found that the regions which significantly differentiated the two groups during learning of B were in the ventrolateral PFC (bilaterally): there were sharp decreases in rCBF here in the 5.5 group but not in the 10-min group. At 5.5 h motor learning again involved the striatum, but this time in the left caudate. Neither the caudate nor the ventral PFC had exhibited learning related activity in field A. Instead, they showed changes in rCBF during the reversal of the learning problem when the previously acquired motor memory was successfully gated. The results demonstrate that: (1) perseveration of a competing motor memory may be linked to reactivation of the neural circuit that participated in acquiring that memory, and (2) the ventral PFC may play an important role in the inhibitory control of the competing motor memory. PMID- 10369147 TI - Rectal temperature and prostaglandin E2 increase in cerebrospinal fluid of conscious rabbits after intracerebroventricular injection of hemoglobin. AB - Fever accompanies subarachnoid hemorrhage (SAH) in the majority of patients. In a previous study, hemoglobin (Hb) was shown to catalyze in vitro, under aerobic conditions, the conversion of arachidonic acid to prostaglandin E2 (PGE2) and prostaglandin F2alpha. The aim of the present work was to assess whether this pathway also operates in vivo and to provide a mechanism to explain post-SAH fever. To this end, PGE2 concentration was determined in cerebrospinal fluid (CSF) of conscious rabbits chronically cannulated in the lateral ventricle and cisterna magna, following intracerebroventricular (i.c.v.) injection of 10 microg or 100 microg of commercial rabbit bicrystallized Hb as a model of SAH. Before i.c.v. injection, Hb solutions were filtered on a polimixin-B column to remove substantially, by over 90%, endotoxin-like substances. Results show that in nine rabbits injection of 10 microg Hb did not significantly modify body temperature or significantly alter CSF PGE2 content. On the contrary, in nine rabbits, injection of 100 microg Hb produced a significant increase in core temperature which was accompanied by a significant increase in CSF PGE2. When data related to these two parameters from the 9 control and 18 Hb-treated rabbits were analyzed as a single group, a linear, positive, and highly significant correlation was found. These findings indicate that, once Hb is released into the subarachnoid space during SAH, it enhances CSF PGE2 content and elicits hyperthermia, thus offering an explanation for the fever that is an aggravating condition in most SAH patients. PMID- 10369148 TI - Comparative study of the neuronal plasticity along the neuraxis of the vibrissal sensory system of adult rat following unilateral infraorbital nerve damage and subsequent regeneration. AB - The aim of the present study was to examine the physiological consequences of a unilateral infraorbital nerve lesion and its regeneration at different levels of the somatosensory neuraxis. In animals whose right infraorbital nerve had been crushed, a large unresponsive area was found in the main brainstem trigeminal nucleus (Pr5). Responses evoked by ipsilateral vibrissal deflection in the middle of Pr5 reappeared only on days 22-35 after the nerve had been transected, whereas recovery from the nerve crush took only 7-9 days. However, no sign of short-term neuronal plasticity was observed in Pr5 after peripheral nerve injury. An enlargement of the receptive fields in two-thirds of the units and a lengthening in the delay of the evoked responses were observed as long-term plastic changes in Pr5 neurons after peripheral-nerve regeneration. In the ventral posteromedial nucleus of the thalamus (VPM) of partly denervated animals, however, only minutes or hours after the nerve crush, certain units were found to respond in some cases not only to the vibrissae, but also to mechanical stimulation of the face over the eye (two units), the nose (one unit), and the midline (one unit). Apart from the experiments involving incomplete denervation, the vibrissal representation areas of the VPM were unresponsive to stimulation of both the vibrissae and other parts of the face until nerve regeneration had occurred. In the somatosensory cortex, an infraorbital nerve crush immediately resulted in a large cortical area being unresponsive to vibrissal deflection. It was noteworthy, however, that shortly after the nerve crush, this large unresponsive whisker representation cortical area was invaded from the rostromedial direction by responses evoked by stimulation of the forepaw digits. In spite of the reappearance of vibrissa evoked responses 7-10 days after the nerve crush, an expanded digital representation could still be observed 3 weeks after the nerve crush, resulting in an overlapping area of digital and vibrissal representations. The withdrawal of the expanded representation of forepaw digits was completed by 60 days after the nerve crush. The results obtained in Pr5, the VPM, and the cortex strongly suggest that the higher the station in the neuraxis, the greater the degree of plasticity after infraorbital nerve injury. PMID- 10369150 TI - Thought disorder in schizophrenia. Testing models through confirmatory factor analysis. AB - Theoretical and empirical models of thought disorder (ThD) were tested through Confirmatory factor analysis (CFA). A sample of 253 DSM-III-R acute schizophrenic patients consecutively admitted was studied. A semistructured interview for schizophrenia was used for diagnosis, and ThD was assessed by means of the Thought, Language, and Communication scale (TLC). Nine ThD models comprising the 18 symptoms of the TLC were tested (ranging from a null model to a six-factor model). The six-dimension model achieved the best fit to the data, although no perfect fit was found. ThD dimensions included in this model were Disorganization, Negative, Idiosyncratic, Semantic, Attentional, and Referential dimensions. The TLC was close to capture adequately these underlying constructs. The Disorganization and the Negative dimensions received more validity on conceptual and psychometric grounds than the remaining dimensions. Thought Disorder multidimensional models fitted the data better than one-dimension models. Thought Disorder dimensions would be potential markers for biological, neurophysiological, and neuropsychological studies of schizophrenic disorder. PMID- 10369151 TI - Prevalence of bipolar II disorder in atypical depression. AB - The diagnostic validity of atypical depression is based on its superior response to monoamine oxidase inhibitors compared to tricyclic antidepressants, and on latent class analysis. The studies on atypical depression have often not included bipolar patients. The aim of the present study was to find the prevalence of bipolar II disorder among DSM-IV atypical depression outpatients. Bipolar II and unipolar atypical depressions were also compared to find if they were variants of the same disorder or if instead they were different disorders. One hundred and forty consecutive unipolar and bipolar II outpatients, presenting for treatment of an atypical major depressive episode, were interviewed with the Structured Clinical Interview for DSM-IV, the Montgomery Asberg Depression Rating Scale (MADRS), and the Global Assessment of Functioning Scale. The prevalence of bipolar II disorder was 64.2%. The age at baseline and onset were significantly lower in bipolar II versus unipolar patients. All the other variables (MADRS items, duration of illness, severity, gender, psychosis, comorbidity, chronicity, recurrences) were not significantly different. The prevalence of bipolar II disorder among atypical depressed outpatients was higher than previously reported. PMID- 10369149 TI - Changes in electrocortical power and coherence in response to the selective cholinergic immunotoxin 192 IgG-saporin. AB - Changes in brain electrical activity in response to cholinergic agonists, antagonists, or excitotoxic lesions of the basal forebrain may not be reflective entirely of changes in cholinergic tone, in so far as these interventions also involve noncholinergic neurons. We examined electrocortical activity in rats following bilateral intracerebroventricular administration of 192 IgG-saporin (1.8 microg/ventricle), a selective cholinergic immunotoxin directed to the low affinity nerve growth factor receptor p75. The immunotoxin resulted in extensive loss of choline acetyl transferase (ChAT) activity in neocortex (80%-84%) and hippocampus (93%), with relative sparing of entorhinal-piriform cortex (42%) and amygdala (28%). Electrocortical activity demonstrated modest increases in 1- to 4 Hz power, decreases in 20- to 44-Hz power, and decreases in 4- to 8-Hz intra- and interhemispheric coherence. Rhythmic slow activity (RSA) occurred robustly in toxin-treated animals during voluntary movement and in response to physostigmine, with no significant differences seen in power and peak frequency in comparison with controls. Physostigmine significantly increased intrahemispheric coherence in lesioned and intact animals, with minor increases seen in interhemispheric coherence. Our study suggests that: (1) electrocortical changes in response to selective cholinergic deafferentation are more modest than those previously reported following excitotoxic lesions; (2) changes in cholinergic tone affect primarily brain electrical transmission within, in contrast to between hemispheres; and (3) a substantial cholinergic reserve remains following administration of 192 IgG-saporin, despite dramatic losses of ChAT in cortex and hippocampus. Persistence of a cholinergically modulated RSA suggests that such activity may be mediated through cholinergic neurons which, because they lack the p75 receptor, remain unaffected by the immunotoxin. PMID- 10369152 TI - Differential therapy effects of psychoeducational psychotherapy for schizophrenic patients--results of a 2-year follow-up. AB - There is increasing evidence of the efficacy and effectiveness of psychosocial interventions in schizophrenic patients. However, little research has been done on differential therapy effects. In a prospective, randomized clinical trial we carried out psychoeducational medication management training, cognitive psychotherapy, and key-person counseling. The patients of the control group participated in structured free-time activities for control of therapeutic commitment. Data from a total of 156 schizophrenic patients (DSM-III-R, no first admissions) were available at 2-year follow-up. We analyzed in this study whether there are differential therapy effects of these interventions, depending on patient characteristics at baseline. There was a significant statistical interaction between treatment condition (specific/non-specific) and prognosis with respect to treatment outcome. Patients with a favorable prognosis and better social functioning had a better course under the specific treatment but a less favorable outcome in the non-specifically treated control group. These results suggest that more vulnerable patients are not sufficiently capable of learning and using coping strategies for relapse prevention. We need to learn more about differential indications for psychosocial treatment. PMID- 10369153 TI - Psychotic relapse and maintenance therapy in paranoid schizophrenia: a 15 year follow up. AB - In spite of numerous reports on a 1 to 2 year maintenance neuroleptic treatment of schizophrenia, there is little systematic information on decade-long maintenance therapy. We conducted a retrospective study in fifty outpatients with paranoid schizophrenia who have been seen at our clinic for a duration of 15 years or more since their first psychotic episodes. Relapse rate within 2, 5, 10, and 15 years from remission of the first psychotic episode were 52, 60, 86, and 90%, respectively. However, the incidence of relapse decreased with time. This decrease was accounted for by the decrease of relapse observed when off drug. Conversely, the incidence of relapse occurred on drug remained unchanged. The average maintenance dose 15 years after remission of the first psychotic episode was 5.41 +/- 7.28 mg/d (haloperidol equivalents: mean +/- SD). The maintenance dose correlated significantly with the number of relapses and total duration of psychotic episodes. These results suggest that maintenance treatment remained effective for decades, although it did not ameliorate the liability to relapse itself. Repeated relapse may be associated with requirement for a higher neuroleptic dose for relapse prevention. PMID- 10369154 TI - Attentional resources in major depression. AB - Depression appears to interfere more with effortful processes than with automatic processes. This study aimed to examine attentional resources allocation by means of RT on effortful detection tasks. Ten depressed inpatients during illness and at recovery and ten healthy control subjects were given simple and choice reaction time tasks. Two types of effort demanding conditions were assessed (1) the combination of two concurrent tasks and (2) tasks involving decision making. Depressed patients improved from single to dual tasks whereas recovered and control worsened. Depressed patients showed a significant time and accuracy impairment when decision processes were involved. The decision making impairment co-occurred with a deficit in the orientation of the attention. The decline with decision making was not worsened when the choice task combined with a concurrent task and was reversible with recovery. This pattern of results exhibits differential sensitivity between two effortful tasks. Depressives may be able to mobilize resources to complete effortful tasks as far as decision processing is not required. PMID- 10369155 TI - Surviving adversity: event decay, vulnerability and the onset of anxiety and depressive disorder. AB - Knowledge concerning the temporal relationship between adverse experiences and the onset of anxiety and depressive disorders remains sparse despite life stress forming a pivotal component to social, neurological and cognitive science models of their aetiology. In this study two groups of married women were selected through their shared adverse experiences; for one group, the marital partner had recently died, and in the second group, the marital partner had recently experienced a myocardial infarction. These groups were assessed in close proximity to their event experiences and again approximately 3 months later. Adaptations of both the Longitudinal Interval Follow-up Evaluation and the Life Event and Difficulty Schedule were used to provide a detailed clinical and event history both preceding and following their experiences. Analysis showed clear evidence for the progressive decay in the adverse effects of life events over time; an attribute thus far largely neglected in work seeking to clarify event illness relationships. Comparisons between fixed and time-varying effects, representative of precisely formulated models of vulnerability/resilience, confirmed the role both of previous psychiatric consultation history and of limited individual coping skills as risk factors for the onset of diagnosable disorder. Improvements in the specification of stress modelling procedures should facilitate the integration of ideas from competing aetiological models of the onset and subsequent course of anxiety and depressive disorder. PMID- 10369156 TI - Psychopathological changes and cognitive impairment in encephalomyelitis disseminata. AB - Two hundred and twelve patients with clinically evidenced encephalomyelitis disseminata (ED), hospitalized in a neurological hospital, were observed with regard to psychopathological characteristics and cognitive changes in conformity with ICD-10 diagnostic criteria. The basis of this investigation was a standardized psychiatric interview. The age of the patients averaged 47 years whereas the duration of the disease averaged 14.3 years. 83.5% of the patients had a disease history of more than 6 years. The medium range of EDSS scores was 5.95%, the BPRS 36.7%. In 5.2% of the patients the course of ED was primarily chronic-progressive while 48% suffered from the intermittent, incomplete reversible form: 47.6% developed secondary chronic-progressive symptoms. 18 psychopathological symptoms could be identified, the main symptom was depressive mood (49%), followed by impairment of affective sensitivity (34.9%) and affective instability/incontinence (31.1%). The most prevalent diagnoses were dementia (23.1%), organic personality disorder (18.5%), mild cognitive impairment (9%), and depressive disorder (7.6%) Only 33.5% were psychopathologically unaffected. The duration of the disease in all demented patients exceeded 6 years. Patients with an organic personality disorder showed a marked increase in the later stages of their illness in contrast to patients suffering from depressive disorder. At the beginning of ED, a highly significant (p < 0.0001) impairment of vision was found in all psychiatric patients. Dementia patients and organic personality patients, on the other hand, showed an advanced degree of ataxia. Actually, there was a considerably lesser incidence of pareses in the non-psychopathological group whereas ataxia was significantly more prevalent in the three cognitively impaired ED-subgroups than in the control group. These findings set the stage for constructive discussions, taking due consideration of existing research results on ED with particular reference to the implications regarding future research as well as the clinical therapy of patients. PMID- 10369157 TI - Spontaneous recurrence of methampetamine psychosis: increased sensitivity to stress associated with noradrenergic hyperactivity and dopaminergic change. AB - We studied the factors precipitating spontaneous recurrences of methamphetamine (MAP)-induced paranoid-hallucinatory states (referred to as "flashbacks") in 28 flashbackers, along with 18 non-flashbackers with a history of MAP psychosis. Plasma levels of catecholamines and their metabolites were assayed in the 28 flashbackers, the 18 non-flashbackers, 8 subjects with persistent MAP psychosis, and 33 normal controls (22 MAP users and 11 non-users). The flashbackers had been exposed to significantly higher numbers of stressful events, and/or MAP-induced frightening paranoid-hallucinatory states during previous MAP use, than the non flashbackers. Factors triggering the flashbacks met the DSM-III-R criteria for a mild psychosocial stressor. During flashbacks, plasma norepinephrine levels increased and plasma levels of 3-methoxytyramine, which is an indicator of dopamine release, showed a smaller increase. It follows that stressful experiences together with MAP use may induce sensitization to mild psychosocial stressors. Noradrenergic hyperactivity and some degree of increased dopamine release may be involved in this process. Stress sensitization may elicit memories of MAP psychosis associated with stressful experiences in response to mild psychosocial stressors, leading to the occurrence of flashbacks. Sensitization to stress associated with noradrenergic hyperactivity, involving increased dopamine release may be central to spontaneous recurrences of MAP psychosis. PMID- 10369158 TI - Two novel polymorphic sequences in the glucocerebrosidase gene region enhance mutational screening and founder effect studies of patients with Gaucher disease. AB - Gaucher disease, an inherited glycolipid storage disorder, is caused by a deficiency of the catabolic enzyme glucocerebrosidase (EC 3.2.1.45). The gene for human glucocerebrosidase is located on chromosome 1q21 and has a highly homologous pseudogene situated 16 kb downstream. We report two novel polymorphic sequences in the glucocerebrosidase gene region: the first consists of a variable number of dinucleotide (CT) repeats located 3.2 kb upstream from the glucocerebrosidase gene, and the second is a tetranucleotide (AAAT) repeat found between the glucocerebrosidase gene and its pseudogene, 9.8 kb downstream from the functional gene. These polymorphic sequences, along with a previously reported PvuII polymorphism in intron 6 of the glucocerebrosidase gene, were analyzed in patients with Gaucher disease (n=106) and in two normal control populations, one of Ashkenazi Jewish ancestry (n=72) and the second comprising non-Jewish individuals (n=46). In these samples, strong linkage disequilibrium was found between mutations N370S, c.84-85insG, and R463C and specific haplotypes; no significant linkage disequilibrium was found when examining haplotypes of patients with the L444P mutation. Studies of these polymorphic sites in several instances also led to the recognition of genotyping errors and the identification of unusual recombinant alleles. These new polymorphic sites provide additional tools for mutational screening and founder effect studies of Gaucher disease. PMID- 10369159 TI - The STR polymorphisms in intron 8 may provide information about the molecular evolution of RH haplotypes. AB - We identified simple-sequence repeat polymorphisms in intron 8 of the RHD and RHCE genes, both of which contained the 5-bp repeat unit (AAAAT)n. We analyzed the polymorphisms of this short tandem repeat (STR) in 104 Japanese RhD-positive and 124 RhD-negative (87 RHD gene negative and 37 nonfunctional RHD gene positive) donors by the polymerase chain reaction (PCR) and subsequent typing by electrophoresis and silver staining. We found five alleles (10, 11, 12, 13, and 14 repeats) in the RHD gene and four (7, 8, 9, and 10 repeats) in the RHCE gene. The Rh phenotypes were closely associated with polymorphisms of the STR. The Ce allele had 12 repeats in the RHD gene and 9 repeats in the RHCE gene at high frequency. The cE allele frequently had 10-12 repeats in the RHD gene and 10 repeats in the RHCE gene. The 10 repeats in the RHCE gene were identified exclusively in the 87 RHD gene-negative donors and 9 repeats were identified only in those with the RhC antigen. These results indicate that both haplotypes of dce and dcE arose from single RHD gene deletion and recombination events, respectively. In the 37 RhD-negative donors with a nonfunctional RHD gene, 12 repeats in the RHD gene and 9 repeats in the RHCE gene were frequently observed. Thus, the RhD-negative with a nonfunctional RHD gene combination might have arisen from the DCe haplotype via a mutation that abolished RHD gene expression. These findings suggest that the STR polymorphisms might shed light upon the molecular evolution of RH haplotypes. PMID- 10369160 TI - Genetic variations in human fetal globin gene microsatellites and their functional relevance. AB - Short tandem repeats are abundantly present within the genome. They are commonly used as polymorphic markers but their potential functional role is poorly documented. Several of these microsatellites have been described within the beta globin locus and some could be involved in controlling gene expression. Our purpose was to investigate the extent and significance of the (TG)n(CG)m dinucleotide repeat polymorphisms in the two gamma-globin gene IVS2s. Two groups of subjects were studied: a group of 63 beta-thalassaemic patients presenting either with a severe Cooley's anaemia (n=50) or with thalassaemia intermedia (TI, n=13), and a control group of 60 unrelated healthy individuals. A high heterogeneity of the polymorphic repeats was demonstrated, extending the range of the published alleles from 13 to 22 and allowing a first attempt at making a phenotype/genotype correlation. One specific allele, (TG)13 in the Agamma-gene, was highly enriched in the TI patients (46.1% vs 2.9% of the Cooley's anaemia cases, P < 0.0002, and 23.3% in the normal controls, P < 0.008) and preferentially observed in TI patients with a high haemoglobin F (Hb F). Transient transfection assays in K562 cells, with the growth hormone gene as a reporter, showed a positive regulatory action mediated by a (TG)13-containing 243 nt IVS2 fragment. Finally, a first set of mobility shift experiments with erythroid (K562 and MEL) and nonerythroid (HeLa) cell lines showed binding of erythroid component(s) in this DNA region and the binding pattern was modified upon induction of MEL cells by DMSO. Thus, our in vivo and in vitro data raise the question of a possible contribution of the gamma-gene IVS2s polymorphic microsatellites to the variable Hb F synthesis in the major haemoglobinopathies: a well known, puzzling and still unanswered question. PMID- 10369161 TI - The structure and dynamics of ring chromosomes in human neoplastic and non neoplastic cells. AB - Acquired ring chromosomes have been found in most types of human neoplasia, with a frequency approaching 10% in malignant mesenchymal tumours. In this study, the composition and dynamics of ring chromosomes were analysed in eight cases of acute myelogenous leukaemia, 17 solid tumours, and five cases with constitutional rings. Chromosomal banding and fluorescence in situ hybridisation were performed to determine the content and the structural heterogeneity of the rings. Telomeric repeats were detected using peptide nucleic acid probes or primed in situ labelling, whereas centromeric activity was evaluated by detection of kinetochore proteins. Mitotic instability was assessed by the frequency of anaphase bridges. The results suggest that human ring chromosomes can be structurally and functionally divided into two categories. In the first of these, size variation is minimal and rearrangement at cell division is uncommon. The majority of such rings contain subtelomeric sequences. Constitutional ring chromosomes and most rings in leukaemias belong to this group, whereas only a few mesenchymal tumours exhibit rings of this type. The second category consists of rings with amplified sequences, primarily from chromosome 12, characteristically occurring in atypical lipomatous tumours and other subtypes of low or borderline malignant mesenchymal neoplasms. Variation in size and number is extensive, and breakage-fusion-bridge events occur at a high frequency. Abnormalities in pericentromeric sequences are common and, in some cases, kinetochores assemble in the absence of alphoid DNA. We conclude that it is not only the ring structure per se or the neoplastic nature of the host cell that determines ring instability, but probably also the functional role of the genes carried in the ring. PMID- 10369162 TI - The human neuregulin-2 (NRG2) gene: cloning, mapping and evaluation as a candidate for the autosomal recessive form of Charcot-Marie-Tooth disease linked to 5q. AB - Neuregulin-2 (NRG2) is a novel member of the neuregulin family of growth and differentiation factors. Through interaction with the ErbB family of receptors, neuregulin-2 induces the growth and differentiation of epithelial, neuronal, glial and other types of cells. In this study, we have cloned the human neuregulin-2 gene, and determined its genomic structure and alternative splicing patterns. By using radiation hybrid mapping panels, the human NRG2 gene was mapped to the D5S658-D5S402 region within 5q23-q33, close to an autosomal recessive form of demyelinating Charcot-Marie-Tooth (CMT) disease. The NRG2 gene was found to be on two yeast artificial chromosomes overlapping the candidate interval and was, thus, considered a good positional candidate for this form of CMT. When the entire neuregulin-2 coding sequence and splice junctions were explored, however, no mutation was identified in one CMT family linked to 5q23 q33. In addition, three intronic single nucleotide polymorphisms were identified in the NRG2 gene. Genotyping in two families localized the NRG2 gene outside of the revised candidate interval between D5S402-D5S210 and excluded NRG2 as the gene responsible for this form of CMT disease. PMID- 10369163 TI - The spectrum of microsatellite loci on chromosomes 7 and 8 in Taiwan aboriginal populations: a comparative population genetic study. AB - Sixteen microsatellite loci on chromosomes 7 and 8 of Han-Taiwanese and six Taiwan aboriginal populations were systematically analyzed by a high-resolution multiple-fluorescence-based polymerase chain reaction technique. Analysis of allele frequency distribution indicated the genetic divergence among these populations. Several alleles were unique to specific tribes. Only the D8S556 locus deviated from Hardy-Weinberg equilibrium in all tribes. Its F(IS) level, as calculated with the Nei method, was also higher and more homozygous than expected. Therefore, with the exception of D8S556, these variable number of tandem repeats (VNTR) loci are suitable genetic markers for forensic and paternal testing. The F(ST) level, as the proportion of the total variation among these tribes, ranged from 1.4% at the D7S484 locus to 6.8% at the D7S550 locus. The average F(ST) was 3.9%, suggesting that there were substantial variations among these populations. The genetic identity analysis and the genetic distance analysis reached the same conclusions, viz., that the Ami and the Paiwan tribes were genetically close to each other, that the Atayal tribe was relatively unique compared with other tribes, and that the Saisiat tribe was relatively close to the Han-Taiwanese. A dendrogram for these tribes was further constructed by the UPGMA method. These VNTR data not only facilitate forensic and paternity testing, but also provide anthropometric information for further elucidating the relationship of Taiwan populations to the Austronesian family. PMID- 10369166 TI - Testing the nonrandomness of chromosomal breakpoints using highest observed breakages. AB - To determine whether a chromosomal band is a fragile site rather than a spontaneous breakpoint, an essential step is to test the nonrandomness of breakage at the region. In this paper, the nonapplicability of the testing procedure introduced by Bohm et al. is discussed, and a new detection procedure is proposed. This new procedure considers the relations of one site with the others, and can be applied to tests of the nonrandomness of breakpoints under either the proportional probability model, or the equiprobability model. A data set for Chinese patients with colorectal carcinoma is analyzed as an illustration of the proposed method. PMID- 10369164 TI - A novel mutation of the doublecortin gene in Japanese patients with X-linked lissencephaly and subcortical band heterotopia. AB - The doublecortin (DCX) gene was recently found to be involved in patients with X linked lissencephaly and subcortical band heterotopia or double cortex syndrome. We have studied the coding regions of the DCX gene in 11 Japanese patients with cortical dysplasia and have identified three different mutations (R186C in exon 3, R272X and R303X in exon 5) in four sporadic female cases. R272X, which has been detected in two unrelated cases, is a novel mutation. Although the number of cases studied remains limited, exon 5 may be a common mutational site in Japanese patients in contrast to many previous reports concerning exons 2 and 3. PMID- 10369165 TI - Analysis of sex chromosome aneuploidy in sperm from fathers of Turner syndrome patients. AB - Numerical sex chromosome abnormalities were analyzed in sperm from four fathers of Turner syndrome patients of paternal origin to determine whether there was an increased frequency of sex chromosome aneuploidy and to elucidate whether meiotic malsegregation mechanisms could be involved in the origin of Turner syndrome. Determination of the parental origin of the single X chromosome (maternal in all four cases) and exclusion of X and Y mosaicism were carried out by polymerase chain reaction amplification of five X chromosome polymorphisms and three Y chromosome segments. A total of 45,299 sperm nuclei from Turner fathers and 85,423 sperm nuclei from eight control donors was analyzed by three-color fluorescence in situ hybridization. The four patients showed a significant increase in the percentages of XY sperm (mean 0.22%; range 0.20% to 0.22%) compared with control donors (mean 0.11%; range 0.06% to 0.18%). These results suggest that the four individuals have an increased frequency of nondisjunctional errors in meiosis I, resulting in the production of an increased proportion of XY spermatozoa and of sperm lacking a sex chromosome. PMID- 10369167 TI - Spinal shock and brain death': somatic pathophysiological equivalence and implications for the integrative-unity rationale. AB - The somatic pathophysiology of high spinal cord injury (SCI) not only is of interest in itself but also sheds light on one of the several rationales proposed for equating 'brain death' (BD) with death, namely that the brain confers integrative unity upon the body, which would otherwise constitute a mere conglomeration of cells and tissues. Insofar as the neuropathology of BD includes infarction down to the foramen magnum, the somatic pathophysiology of BD should resemble that of cervico-medullary junction transection plus vagotomy. The endocrinologic aspects can be made comparable either by focusing on BD patients without diabetes insipidus or by supposing the victim of high SCI to have pre existing therapeutically compensated diabetes insipidus. The respective literatures on intensive care for BD organ donors and high SCI corroborate that the two conditions are somatically virtually identical. If SCI victims are alive at the level of the 'organism as a whole', then so must be BD patients (the only significant difference being consciousness). Comparison with SCI leads to the conclusion that if BD is to be equated with death, a more coherent reason must be adduced than that the body as a biological organism is dead. PMID- 10369168 TI - Family situation and psychosocial issues including problems that impact on partnership or resettlement in the home and into society. PMID- 10369169 TI - Diurnal variation of antidiuretic hormone and urinary output in spinal cord injury. AB - INTRODUCTION: Healthy individuals have a nocturnal decrease in urine output due to increased plasma antidiuretic hormone levels at night. This does not occur in spinal cord injury and most patients experience nocturnal polyuria, which triggers dysreflexic crises secondary to urinary bladder overdistension, and interferes with patients' sleep due to the need for extra catheterization. OBJECTIVE: To evaluate the diurnal variation in ADH level, urinary output, and plasma and urine osmolality in SCI patients with regard to their level of injury and in comparison with age- and sex-matched healthy individuals. MATERIALS AND METHODS: Sixteen ASIA-A spinal cord-injured patients, eight with paraplegia, eight with tetraplegia, and eight healthy individuals, were evaluated for urinary output, urine and serum osmolality, and antidiuretic hormone levels during day and night hours. RESULTS: Absence of diurnal variation in urinary output and antidiuretic hormone secretion was detected in both paraplegic and tetraplegic patients, while antidiuretic hormone levels rose significantly at night in the control group. CONCLUSION: Antidiuretic hormone levels should be monitored both day and night in spinal cord injury patients, with severe nocturnal polyuria. Treatment with desaminocystein-D-arginine vasopressin can be attempted when conservative measures fail to control nocturnal polyuria, especially in patients who are on an intermittent catheterization program. PMID- 10369170 TI - Ambulation training of neurological patients on the treadmill with a new Walking Assistance and Rehabilitation Device (WARD). AB - STUDY DESIGN: Patients with neurological walking impairment were rehabilitated with a new system, consisting of an apparatus to constantly relieve the body weight and a treadmill: The Walking Assistance and Rehabilitation Device (WARD). Patients were evaluated before and after rehabilitation with clinical scales and physiological measurements. OBJECTIVES: To evaluate the effectiveness of the WARD in improving walking capability in these patients. SETTING: The study was carried out in a clinical environment (IRCCS S. Lucia, Rehabilitation Hospital, Rome, Italy). METHODS: Seven patients (six with spinal cord injuries, one with brain injury) underwent a 1 - 2 month training period with the WARD. During the WARD training the body weight constant unloading (BWCU) applied to the patient through the WARD was regularly evaluated. Oxygen consumption, carbon dioxide production and heart rate were measured in order to obtain energy and cardiac costs of walking. These measurements were carried out while walking with the WARD at an appropriate treadmill speed (ATS) and in the open field at the most comfortable speed (MCS). All measurements, in addition to clinical scores related to the walking capability, were carried out at the beginning of the WARD training period (BWT) and at the end (EWT). At the EWT the patients were tested walking with the WARD observing the same BWT conditions (same ATS and BWCU), referred to as beginning conditions second measurements (BCSM). The relationships between physiological costs and ATS were described through second order polynomial regression curves and studied. RESULTS: Comparing the data obtained at the BWT and EWT, the following results were found significantly different: (1) Clinical scores improved; (2) All patients increased their ATS; (3) The BWCU was reduced; (4) The Walking Energy Cost (WEC) and the Walking Cardiac Cost (WCC) measured when walking with the WARD at the ATS improved in all patients; and (5) The WEC and WCC measured in the open field improved in all patients. The WEC vs ATS curve found at the EWT in the BCSM was found significantly different from the BWT curve, demonstrating a major improvement due to the WARD training. CONCLUSION: Despite some limitations due to sample size and functional ambulation scale, this study has demonstrated that the WARD training is effective in improving the walking capability and efficiency of the patients. PMID- 10369172 TI - Neurological abnormalities, major orthopaedic deformities and ambulation analysis in a myelomeningocele population in Catalonia (Spain). AB - The aim of the study was to analyze the present status of neurologic abnormalities, major orthopaedic deformities and ambulatory status in a large myelomeningocele population. PATIENTS AND METHODS: Cross-sectional study based on the clinical and radiographic records of 322 patients treated and followed-up from 1967-1995. The setting was a multidisciplinary spina bifida unit within a third-level university hospital, which serves as the referral centre for these patients in Catalonia (Spain). We collected information on diagnosis, central nervous system, musculoskeletal system (spinal and hip deformities) and functional level in each patient. To study relationships among the variables, the Mann-Whitney U and the Chi-squared tests were applied. Results were considered to be statistically significant at P levels of < or = 0.05. RESULTS: Mean age was 15.9 years. 78.1% of patients had mid-lumbar, low-lumbar or sacral neurological levels; 97.5% had hydrocephalus and 68.8% were shunted. Prevalence of spine deformities was 45.3%; 38.8% had dislocation of one or both hips. Median age of walking onset was 37.1 months and 74.8% of patients were ambulatory. Median age at which ambulation ceased was 128 months (10 years and 8 months). The bivariate analysis showed statistically significant relationships between neurological level and all the variables studied (P<0.001, P<0.02) except body mass indexes and intelligence quotient. CONCLUSIONS: Neurological level was the main factor that determined neurological abnormalities, major orthopaedic deformities and ambulatory status. PMID- 10369171 TI - Improving the long-term adjustment of spinal cord injured persons. AB - STUDY DESIGN: The study involved the long-term assessment of persons with spinal cord injury (SCI) who previously participated in a nonrandomized longitudinal controlled trial. OBJECTIVES: The objective was to report on the effectiveness of early psychological intervention in improving some aspects of adjustment to SCI 2 years post injury. SETTING: The study was conducted in Sydney, Australia. METHODS: Twenty-eight SCI persons who had participated in group Cognitive Behaviour Therapy (CBT) during hospital rehabilitation were assessed for drug usage, hospital readmissions, relationships, perceived adjustment and social discrimination 2 years after treatment. The intervention group's responses on the measures were compared with a control group of 31 SCI persons who only received traditional rehabilitation services during their hospitalization. RESULTS: Subjects in the treatment group experienced less hospital re-admissions, used less drugs and reported higher levels of adjustment compared to the control group. Relationships were stable for both groups and there were no suicides in either group up to 2 years. There were no differences in perceived social discrimination between the two groups. CONCLUSION: The study suggests the long term adjustment for SCI persons is encouraging for the majority. However, the provision of group CBT for SCI persons appears to improve significantly some aspects of adjustment to the injury in the long-term. PMID- 10369173 TI - Bone mineral density differences between paraplegic and quadriplegic patients: a cross-sectional study. AB - STUDY DESIGN: This cross-sectional study was conducted by comparing bone mineral density (BMD) of paraplegic and quadriplegic patients. OBJECTIVES: The purpose of this study was to investigate the relationship between the bone mineral loss and injury level in spinal cord injury patients. SETTINGS: Experiments were conducted at Yoneda Hospital and Research Center of Health, Physical Fitness and Sports, Nagoya University, Nagoya, Japan. METHODS: Lumbar spine (L2-4), proximal femur (femoral neck, trochanter region and Ward's triangle) and whole body BMD were measured in ten paraplegic and ten quadriplegic patients using dual-energy X-ray absorptiometry (DXA, HITACHI BMD-IX). RESULTS: Significant differences were observed in the lumbar spine, trochanter region and upper extremities BMD between paraplegic and quadriplegic patients (P<0.05, P<0.05 and P<0.01, respectively), but not in the femoral neck, Ward's triangle, head, pelvis, lower extremities or whole body BMD. CONCLUSION: These results suggest that the injury level influences on the lumbar spine, upper extremities and trochanter region BMD. From a biomechanical standpoint, it is possible to explain that the differences in mechanical loading exerted on bones also affected the difference of lumbar spine BMD in the two groups. PMID- 10369174 TI - Tuberculosis of spine (C1 to D4). AB - CLINICAL DATA: Thirty-one patients with 33 lesions of spinal tuberculosis (C1-D4) are reported. The distribution of lesions was C1-C2 (11), C3-C6 (13), C7-D4 (9). Neurological complications were present in 6 (55%), 8 (61%) and 7 (78%) in each region respectively. DIAGNOSIS: Increase in the prevertebral soft tissue shadow in a standard radiograph was a useful guide to resort to CT Scan/MRI to diagnose tuberculosis of C1 and C2 region at an early (pre-subluxation) stage. The diagnosis of TB spine from C3-C6 was made confidently on clinico-radiological features. The anterior convexity and forward displacement of tracheal shadow of more than 8 mm from the vertebral bodies in a lateral view of plain X-ray and widening of superior mediastinum in an AP X-ray are useful indicators of tuberculous involvement at cervicodorsal region (C7-D4). CT Scan/MRI should be done for early diagnosis in those cases with a high index of suspicion. TREATMENT AND OUTCOME: 12/33 lesions without neural complications healed with antitubercular drugs and the use of suitable orthosis. Out of 21 lesions with neural complications 14 recovered by local rest, skull traction and multidrug therapy. Seven lesions were surgically decompressed. Of these, five recovered completely, two did not achieve useful recovery. The neural recovery following the middle path regimen for tuberculosis of C1-D4 was 90% in our cases. PMID- 10369175 TI - Regression of vasomotor disorders under intrathecal baclofen in a case of spastic paraplegia. AB - Continuous intrathecal baclofen infusion via a subcutaneously implanted programmable pump has been used in the treatment of severe spasticity. Improvement classically concerns the neurological (hypertonia, spasms, hyperreflexia), urological (bladder function) and other clinically relevant outcomes, such as functional status of daily living. This short note reports on another effect of intrathecal baclofen on vasomotor disorders and cyanosis in the lower limbs, described in a patient with spastic paraplegia. PMID- 10369176 TI - Squamous cell carcinoma of suprapubic cystostomy tract without bladder involvement. AB - This report describes a third case of squamous cell carcinoma of the suprapubic cystostomy tract. The first case reported in 1993 concerned a squamous cell carcinoma arising adjacent to the suprapubic cystostomy site and extending anteriorly to the abdominal wall in a 80-year-old man, 5 years after suprapubic urinary diversion for urethral stricture. A second case published in 1995 described a 50-year-old paraplegic man (T11-T12 spinal cord injury) in whom a suprapubic cystostomy tract squamous cell carcinoma developed after 25 years of urinary diversion. The tumour involved the cystostomy tract primarily with extension into the bladder but did not penetrate the bladder wall muscle. Our patient is in fact the second one to have a suprapubic cystostomy tract squamous carcinoma not involving the bladder. PMID- 10369177 TI - Bladder stones of unusual shape in a male with paraplegia due to spinal cord injury who has been performing self-catheterisation. PMID- 10369178 TI - Thoracolumbar fractures. PMID- 10369180 TI - NO-dependent mechanisms of adaptation to hypoxia. AB - In studying NO-dependent mechanisms of resistance to hypoxia, it was shown that (1) acute hypoxia induces NO overproduction in brain and leaves unaffected NO production in liver of rats; (2) adaptation to hypoxia decreases NO production in liver and brain; and (3) adaptation to hypoxia prevents NO overproduction in brain and potentiates NO synthesis in liver in acute hypoxia. Dinitrosyl iron complex (DNIC, 200 microg/kg, single dose, iv), a NO donor, decreases the resistance of animals to acute hypoxia by 30%. Nomega-nitro-L-arginine (L-NNA, 50 mg/kg, single dose, ip), a NO synthase inhibitor, and diethyl dithiocarbamate (DETC, 200 mg/kg, single dose, iv), a NO trap, increases this parameter 1.3 and 2 times, respectively. Adaptation to hypoxia developed against a background of accumulation of heat shock protein HSP70 in liver and brain. A course of DNIC reproduced the antihypoxic effect of adaptation. A course of L-NNA during adaptation hampered both accumulation of HSP70 and development of the antihypoxic effect. Therefore, NO and the NO-dependent activation of HSP70 synthesis play important roles in adaptation to hypoxia. PMID- 10369179 TI - S-nitrosothiols are stored by platelets and released during platelet-neutrophil interactions. AB - Interaction between platelets and neutrophils is important in vascular injury. We have investigated the storage and release of nitric oxide (NO) by platelets interacting with neutrophils. Shear-activated platelets were added to neutrophils in suspension and both superoxide and peroxynitrite formations monitored by lucigenin- and luminol-enhanced chemiluminescence. In addition, intraplatelet S nitrosothiols were measured by dichlorofluorescein fluorescence following displacement of NO by mercuric chloride. Addition of activated platelets to neutrophils caused free radical production and platelet-neutrophil rosette formation. Pretreatment of platelets with 20 microM S-nitrosoglutathione changed the balance between luminol and lucigenin chemiluminescence in favor of luminol, whereas S-nitrosoglutathione in platelet-free plasma did not produce these changes. This pattern was also observed both following inhibition of neutrophil NO synthase and in a neutrophil-free superoxide-generating system. Inhibition of platelet NO synthase decreased luminol and increased lucigenin chemiluminescence. These effects were reversed by L-arginine. Platelet activation increased intraplatelet S-nitrosothiols from 1.93+/-0.19 (mean +/- SD) to 4.9+/-1.10 x 10( 18) mol/platelet (P < 0.01); this increase halved following NO synthase inhibition, but was enhanced by approximately 220% following incubation with S nitrosoglutathione. These results show that during shear stress platelets store S nitrosothiols, which can be derived either endogenously from NO synthesis or exogenously by sequestration of S-nitrosoglutathione. Release of stored NO during platelet-neutrophil interaction alters the interaction of NO with superoxide and could modulate vascular inflammation. PMID- 10369181 TI - Continuous monitoring of in vivo nitric oxide formation using EPR analysis in biliary flow. AB - A method to continuously monitor the nitric oxide (NO) level in anesthetized rats, using an in vivo trapping reaction of NO by iron-dithiocarbamate complex, is reported. Previously, we developed a method of monitoring NO in bile samples containing an NO complex excreted from the liver (Anal. Biochem. 243, 8-14, 1996). In the present study, we modified the method so that the bile flows directly through the EPR sample cell. Rats were injected with low doses of lipopolysaccharide (LPS) to induce NO formation and were later anesthetized. After cannulation, the bile duct was connected to the inlet of the EPR sample cell and the trapping agent iron complex of D-N-methylglucamine dithiocarbamate (MGD-Fe) was administered. The EPR signal level from NO complex of MGD-Fe in the flowing bile was continuously monitored. Using this method, immediate changes in in vivo NO level in rats were observed following administration of drugs that can affect NO formation. In addition, a continuous intravenous saline containing MGD Fe made the EPR signal level stable and improved animal condition as well as survival time. Therefore, this method has two merits; (1) one can continuously monitor NO formation until it reaches the maximum level; (2) a rapid change in NO level after intervention can be followed. Using this method, we tested the effect of the substrate L-arginine and inhibitors for NO synthase activity and NO synthase induction. The sensitivity of the present method was tested by monitoring NO formation in rats after exposure to ionizing radiation. PMID- 10369182 TI - Stimulation of NO synthase activity in the immune-competent lepidopteran Estigmene acraea hemocyte line. AB - In contrast to the vertebrate immune system, nearly nothing is known about the immunological role of nitric oxide (NO) in invertebrates. This study provides evidence of the presence of a NO synthase (NOS) activity in an immune-competent, macrophage-like insect hemocyte line, previously established from larvae of the lepidopteran insect Estigmene acraea. As proven by photometric determination of nitroblue tetrazolium reduction after cell fixation, the E. acraea cells possess NADPH diaphorase (NADPHd) activity. This NADPH diaphorase activity was NADPH dependent, not inhibitable by superoxide dismutase, influenced by extracellular addition of L-arginine, and inhibited in a dose-dependent manner by the specific NOS inhibitor Nomega-monomethyl-L-arginine. Furthermore, the NADPH diaphorase activity was stimulated within 30 min by the addition of insect pathogenic bacteria (Bacillus thuringiensis var. kurstaki, Photorhabdus luminescens), bacterial lipopolysaccharide, and silica beads. In activated E. acraea cell suspensions strongly increased amounts of L-citrulline and enhanced levels of total nitrite/nitrate (as NO derivates) can be determined. This is the first report on stimulable NOS activity in insect hemocytes. PMID- 10369183 TI - Analysis of 3-nitrotyrosine in biological fluids and protein hydrolyzates by high performance liquid chromatography using a postseparation, on-line reduction column and electrochemical detection: results with various nitrating agents. AB - Nitric oxide reacts rapidly with superoxide to form the strong nitrating agent peroxynitrite, which is responsible for much of the tissue damage associated with diverse pathophysiological conditions such as inflammation. The occurrence of free or protein-bound nitrotyrosine (NTYR) has been considered as evidence for in vivo formation of peroxynitrite. However, various agents can nitrate tyrosine, and their relative significance in vivo has not been determined due to lack of a sensitive method to analyze NTYR in tissue proteins and biological fluids. We have developed a new HPLC-electrochemical detection method to analyze NTYR in protein hydrolyzates or biological fluids. The sample is injected directly into a reversed-phase HPLC column and NTYR is subsequently reduced by a platinum column to 3-aminotyrosine, which is quantified with an electrochemical detector. The method is simple, selective, and sensitive (detection limit, 0.1 pmol per 20 microl injection). We have applied this method to compare in vitro the ability of various nitrating agents to form NTYR in bovine serum albumin and human plasma. Yields of NTYR formed in human plasma proteins incubated with 1 or 10 mM nitrating agent decreased in the following order: synthetic peroxynitrite > 3 morpholinosydonimine, a generator of both NO and superoxide > Angeli's salt, which forms nitroxyl anion (NO-) > spermine-NONOate, which releases NO > sodium nitrite plus hypochlorite, which forms the nitrating agent nitryl chloride (NO2Cl). A simple purification method using a C18 Sep-Pak cartridge is also described for analysis of free NTYR in human plasma. PMID- 10369184 TI - Peroxynitrite reaction with heme proteins. AB - We have reported that low levels of peroxynitrite (PN) can cause inactivation of the heme-thiolate protein prostacyclin (PGI2)-synthase by nitration of a tyrosine residue. To prove that iron catalysis is involved we studied the interaction of PN with microperoxidase and P450nor, a heme-thiolate protein of known structure. Spectral and kinetic analyses allow to conclude on a ferryl nitrogen dioxide complex as an intermediate which decomposes in the presence of an excess of PN under formation of dioxygen, nitrite, and nitrate. This occurs in a catalytic cycle which was more efficient with P450nor than with microperoxidase. If phenol was added to the reaction mixtures of PN and the ferric complexes the ratio of hydroxylated to nitrated phenols decreased compared to the metal-free system. Phenol competed with the formation of dioxygen indicating that the ferryl intermediate was involved in both pathways. One therefore can postulate that the ferryl complex reacts with phenol to give the phenoxyradical which is nitrated in the presence of nitrogen dioxide but does not give hydroxylated products as with metal-free PN. Alternately, the ferryl nitrogen dioxide complex can oxidize a second PN molecule to the radical, *OONO, which can decompose to dioxygen and NO. The latter forms N2O3, with the remaining *NO2 radical. A third pathway consists in the isomerization to nitrate which also is catalyzed by the heme proteins since the ratio of nitrite/nitrate does not change significantly during the catalytic reaction with excess of PN. Our data explain the mechanism of nitration of PGI2-synthase, suggest a role of P450nor as a PN scavenger, and favor heme thiolate complexes for trapping PN. PMID- 10369185 TI - Increased nitric oxide concentrations in posterior hypothalamus and central sympathetic function on nitrate tolerance following subcutaneous nitroglycerin. AB - The present study was to examine the distributions of nitric oxide (NO) in the brain regions and peripheral vessels following subcutaneously administered nitroglycerin (NTG) and determine the noradrenergic activity and the role of central sympathetic function in acute nitrate tolerance. Tolerance to NTG was produced by subcutaneous (sc) administration of 4.0 mg NTG as four separate hourly pulse injections of 1.0 mg each in male (5-8 months) Sprague-Dawley rats. Rats in sham-treated group received sc injections of saline. Rats were killed by sodium pentobarbital (150 mg/kg, ip) at 10 min after last sc injection. The brain, gracilis muscle, aorta, superior mesenteric artery, coronary artery, and pulmonary vessels were quickly removed. Concentrations of nitrite (NO2-), nitrate (NO3-), and total NO2- plus NO3- (NOx-) were quantified in the micropunches of the anterior hypothalamus, the posterior hypothalamus (PH), the nucleus tractus solitarius, the lateral reticular nucleus, and the vessels in a blinded fashion. The central actions of acute tolerance to NTG were also determined using blockades of sympathetic functions in conscious rats. Four separate hourly pulse sc injections of 1.0 mg NTG produced a marked shift of the dose-response curve for arterial pressure depression induced by intravenous injection of the challenge doses of NTG. The same doses of sc NTG caused increases in NOx- [92+/ 16% (mean +/- SE)] and NO3- productions (77+/-15%) in the PH, but did not significantly change in other brain regions (n = 6). NOx- and NO3- productions were significantly enhanced in the superior mesenteric artery, aorta, coronary artery, and pulmonary vessels following sc NTG, but were not altered in gracilis muscle by the treatment. The tolerance responses to arterial pressure depression were attenuated by intravenous administration of either prazosin (300 microg/kg), an alpha1-adrenoceptor antagonist, or chlorisondamine (10 mg/kg), a sympathetic ganglion blockading agent (n = 5-6). The results suggest that acute NTG tolerance predominately increases NO production in the PH. NO production was also markedly enhanced in the large and middle vessels but not in small vessels during acute NTG tolerance. The arterial pressure tolerance to NTG was reversed by blockade of central sympathetic function. We conclude that NO formation is increased in the PH following systemically administered NTG and NO in the PH may facilitate central sympathetic functions which contribute to nitrate tolerance. PMID- 10369186 TI - Evidence for nitric oxide action on in vitro granuloma formation through pivotal changes in MIP-1alpha and IL-10 release in human schistosomiasis. AB - Nitric oxide (NO) has been implicated, both and paradoxically, as a pro- and anti inflammatory agent in a wide range of circumstances. It is of common concern that NO can be either up- or downregulated by different inflammatory cytokines. Attempting to assess the contribution of NO to the granulomatous response, we used the in vitro granuloma (IVG) model which consists on a reaction of mononuclear cells around polyacrylamide beads conjugated to antigens. Our assays employed Schistosoma mansoni antigens and human peripheral blood mononuclear cells (PBMC) from schistosomiasis patients. Recently, we have described evidence for a regulatory role of NO, with the aid of an inhibitor of NO synthesis, L NAME. The addition of L-NAME to IVG cultures elicited an increase on the granuloma formation index. Based on these data we decided to investigate the mechanisms involved in the effects of L-NAME-enhanced granuloma formation. Cytokines and chemokines are involved in inflammatory responses by, particularly the latter, inducing migration and adhesion of leukocytes, which led us on this search for their interactions with NO on granulomatous reaction. We evaluated the cytokine/chemokine-secreting profile of PBMC (treated and not treated with L NAME) on the IVG reaction in order to investigate how NO could interfere on the release of these soluble mediators. Comparison of cell culture releasing amounts of IL-2, IL-10, TNFalpha, IFNgamma, MIP-1alpha, MCP-1, and RANTES demonstrated that MIP-1alpha had increased levels when NO production was blocked with L-NAME, whereas IL-10 secretion decreased in presence of L-NAME. The other tested cytokines (IL-2, TNFalpha, and IFNgamma) and chemokines (MCP-1 and RANTES) showed no significant differences between the presence or absence of L-NAME. Results obtained in this work suggest that inhibition of NO production could upregulate the IVG reaction on human schistosomiasis through changes in the cytokine/chemokine profile released by PBMC. The mechanisms involved may lead to a MIP-1alpha-increased and IL-10-decreased secretion under our experimental conditions, which could partly account for the previously ascribed IVG exacerbating action of NO inhibition. PMID- 10369187 TI - Blockade of nitric oxide synthase potentiates the suppression of vasodilators by norepinephrine in the hepatic artery. AB - We have previously shown that nitric oxide (NO) and adenosine suppress vasoconstriction induced by norepinephrine infusion and sympathetic nerve stimulation in the hepatic artery and superior mesenteric artery. NO is involved in the control of basal vascular tone in the superior mesenteric artery but not the hepatic artery. The vasodilation induced by adenosine is inhibited by NO in the superior mesenteric artery but not in the hepatic artery. Based on these known interactions of catecholamines, adenosine, and NO, the objective of this study was to test the hypothesis that NO modulates the interaction between vasoconstrictors and vasodilators in the hepatic artery. We examined the ability of norepinephrine to suppress adenosine-mediated vasodilation and the role of NO in this interaction. Hepatic arterial blood flow and pressure were monitored in pentobarbital-anesthetized cats. The maximum hepatic arterial vasoconstrictor response to norepinephrine infusion was potentiated by blockade of NO production using Nomega-nitro-L-arginine methyl ester (L-NAME), and the potentiation was reversed by L-arginine. The maximum dilator response to adenosine was only slightly suppressed (14.0+/-5.8%, P < 0.05) by norepinephrine infusion; however, after the NO blockade, the suppression by norepinephrine of the vasodilation induced by adenosine was substantially potentiated (45.2+/-9.1%, P < 0.05). Similar results were obtained for isoproterenol-induced vasodilation. We conclude that the interaction between these vasodilators and norepinephrine was modulated by NO which inhibited the vasoconstriction and the suppression of vasodilators caused by norepinephrine and that in the absence of NO production, norepinephrine induced constriction and the ability to antagonize dilation is substantially potentiated. PMID- 10369189 TI - Evidence-based recommendations for the management of glomerulonephritis. Introduction. PMID- 10369188 TI - Intercellular signaling initiated by nitric oxide produced in heat-shocked human glioblastoma cells. AB - The accumulation of inducible nitric oxide synthase was caused by heat shock of human glioblastoma T98G cells but not of A-172 cells. The accumulation of hsp72 and p53 was observed in A-172 cells cocultivated with heat-shocked T98G cells, which was suppressed by the addition of aminoguanidine to the medium. The accumulation of these proteins was observed in A-172 cells after exposure to the conditioned medium of heat-shocked T98G cells, which was completely blocked by the addition of 2-(4-carboxyphenyl)-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3 oxide to the medium. In addition, the accumulation of these proteins in A-172 cells was induced by the administration of S-nitroso-N-acetylpenicillamine to the medium. Finally, the thermosensitivity of A-172 cells was reduced in the conditioned medium of heat-shocked T98G cells compared with conventional fresh growth medium. Our findings demonstrate that the accumulation of stress-induced proteins and thermoresistance in NO recipient cells cocultivated with heat shocked NO donor cells is induced through an intercellular signal transduction pathway initiated by NO without cell-to-cell interactions such as gap junctions. PMID- 10369190 TI - Management of minimal lesion glomerulonephritis: evidence-based recommendations. AB - The treatment of idiopathic minimal lesion disease in children has been extensively studied in randomized controlled trials, however, there is less information available for adults. This article summarizes evidence-based recommendations for management. The first attack should be treated with prednisone or prednisolone at 60 mg/m2 per day (up to a maximum of 80 mg/day) for four to six weeks, followed by 40 mg/m2 of prednisone every other day for another four to six weeks (grade A). Relapse should be treated with 60 mg/m2/day of prednisone (up to 80 mg/day) only until the urine becomes protein free for three days, and then an alternate day regimen of 40 mg/m2 should be used for another month (grade A). Patients with frequently relapsing disease will have a significant reduction in relapse frequency after eight weeks of an alkylating agent (grade A). Less rigorous studies have suggested benefit with long-term, alternate-day corticosteroid (grade D) or the antihelminthic agent levamisole (grade D). For patients with steroid-dependent disease, an 8- or 12-week course with cyclophosphamide can induce remission (grade D). In true steroid-resistant disease, observational studies have suggested that a course of cyclosporine may sometimes induce remission or restore steroid responsiveness (grade D). Large retrospective studies in adults suggest that therapeutic response is slower than in children, but adults experience fewer relapses and more prolonged remission. PMID- 10369191 TI - Conservative treatment to slow deterioration of renal function: evidence-based recommendations. AB - Despite current specific therapy, progressive deterioration of renal function in patients with primary glomerulonephritis occurs. Nonspecific renoprotective interventions that have been studied include blood pressure control, antihypertensive medications, and protein-restricted diets. To prepare this article, a MEDLINE search was conducted, followed by secondary and tertiary searches. Research papers were assessed for level of evidence, and graded recommendations were formulated. Protein-restricted diets (to 0.4 to 0.6 g/kg/day) are not recommended for all patients with reduced renal function (grade A). Very low-protein diets of 0.4 g/kg/day should be considered for patients with severe renal dysfunction (serum creatinine of more than 350 micromol/liter; grade A). However, there are concerns about recommending these diets for all patients because of the potential for long-term negative outcomes such as nutritional deficiencies. Target blood pressure for persons with proteinuria of more than 1 g/day should be less than 125/75 mm Hg [mean arterial pressure (MAP) < 92 mm Hg; grade C]. For persons with proteinuria of less than 1 g/day, the target blood pressure should be approximately MAP 98 mm Hg (less than 130/80; grade C). Angiotensin-converting enzyme inhibitor (ACEI) therapy is recommended in preference to placebo, conventional, or beta-blocker therapy for renoprotection (grade A). ACEI therapy cannot be recommended above calcium channel blockers in patients with nondiabetic renal disease (grade A). PMID- 10369193 TI - Management of idiopathic crescentic and diffuse proliferative glomerulonephritis: evidence-based recommendations. AB - Idiopathic crescentic glomerulonephritis (GN) often presents with a rapid loss of renal function and pathology showing extensive crescent formation. The disease is caused by different immunopathogenetic mechanisms, pauci-immune, often antineutrophil cytoplasmic antibody (ANCA)-positive microvasculitis, antiglomerular basement membrane (GBM) antibody disease, and immune complex formation. Historical reviews reveal poor renal prognosis, even after treatment with oral steroids and cytotoxic drugs. Prognosis has improved in the last decade. In this article, evidence-based recommendations for management are presented. Because of the high risk of end-stage renal disease (ESRD), early aggressive therapy is recommended, despite weak supporting evidence. Treatment for anti-GBM antibody-induced crescentic GN should be initiated early and should include pulse methylprednisolone, a two-week course of plasmapheresis and two months of treatment with corticosteroids and cyclophosphamide (grade B and C). Treatment for pauci-immune crescentic GN should be pulse methylprednisolone, followed by oral corticosteroids and cyclophosphamide for 6 to 12 months (grade B). Recurrences can be managed similarly (grade B), along with appropriate supportive therapy. In patients who develop ESRD, successful transplantation can be performed. Diffuse endocapillary proliferative GN is classically postinfectious. It generally has a good prognosis when no crescent formation occurs. Adult patients with persistent proteinuria, hypertension, and renal function impairment need careful follow-up and management to modify progressive hemodynamic injury. PMID- 10369192 TI - Management of focal segmental glomerulosclerosis: evidence-based recommendations. AB - Focal segmental glomerulosclerosis (FSGS) is a diagnosis based on the presence of glomeruli with segmental scarring in association with intracapillary foam cells and adhesions. To develop evidence-based treatment guidelines, a MEDLINE search was conducted, and articles were reviewed using levels of evidence. Graded recommendations were developed according to the level of evidence. There was limited evidence found on which to develop recommendations. Treatment with prednisone of 0.5 to 2 mg/kg/day should be considered in all patients and continued for six months before declaring the patient resistant to therapy. Remission is associated with the use of high doses (more than 60 mg/day) for three months; therefore, if there is a concern about prolonged use, a reduction in dose to 0.5 mg/kg/day should be made only after three months (grade D). The use of cyclosporine A (CsA) at doses to maintain serum levels at 150 to 300 microg/ml may be effective in reducing urinary protein excretion. Relapse after reducing or stopping CsA is very common. Long-term use may be required to maintain remission (grade D). The use of cytotoxic therapy (cyclophosphamide, azathioprine, and chlorambucil) for adults is second-line therapy (grade D). Plasmapheresis or protein adsorption may be recommended for renal transplant patients with recurrent FSGS (grade D). PMID- 10369194 TI - Management of membranoproliferative glomerulonephritis: evidence-based recommendations. AB - Idiopathic membranoproliferative glomerulonephritis (MPGN) is one of the least common types of GN. This article critically evaluates the literature and generates evidence-based recommendations for the management of idiopathic MPGN. For all age groups, for idiopathic MPGN with normal renal function and asymptomatic nonnephrotic range proteinuria, no specific therapy is necessary (grades B and C). Close follow-up every three to four months, with specific attention to renal function, proteinuria, and blood pressure control, is recommended. In children with MPGN and nephrotic syndrome and/or impaired renal function, a trial of steroids is warranted (grade A). The best data suggest high dose, alternate-day steroids for a period of 6 to 12 months (40 mg/m2 on alternate days). If no benefit is seen, discontinuation with close follow-up and attention to conservative treatment (that is, blood pressure control, use of agents to reduce proteinuria, and correction of metabolic abnormalities) is recommended. In adults with MPGN, impaired renal function, and/or nephrotic-range proteinuria, a trial of aspirin (325 mg daily), dipyridamole (75 to 100 mg tid), or a combination of the two for 12 months is reasonable (grade B). Again, if no benefits are seen, the treatment should be stopped. Attention to factors known to delay the progression of renal decline and close follow-up should be part of the treatment plan (grades B and C). PMID- 10369195 TI - Management of idiopathic membranous nephropathy: evidence-based recommendations. AB - Membranous nephropathy is a frequent cause of nephrotic syndrome in adults, and in one third of these patients, it leads to end-stage renal disease. Based on an extensive critical review of the literature, the following recommendations are offered. Oral high-dose corticosteroids are ineffective in producing either a sustained remission of nephrotic syndrome or in preserving renal function in patients with membranous nephropathy, and should not be used as the sole therapy (grade A recommendation). The use of azathioprine is not associated with any significant benefits, so its use is not justified (grade C). The alkylating agents cyclophosphamide and chlorambucil are both effective in the management of membranous nephropathy. Because of growing concern about long-term toxicity, especially with cyclophosphamide, these drugs should be reserved for patients who exhibit clinical features, such as severe or prolonged nephrosis, renal insufficiency, or hypertension, that predict a high likelihood of progression to end-stage renal disease. Chlorambucil in conjunction with oral steroids is the drug of first choice (grade A). Cyclophosphamide and oral steroids are alternatives (grade B). Cyclosporine may, in the future, become the agent of choice for membranous nephropathy. Currently, it is recommended (grade B) that cyclosporine use be considered in patients at high risk for progression in membranous nephropathy or if alkylating agents are contraindicated or ineffective. PMID- 10369197 TI - The brain as a new frontier for reproductive endocrinology. PMID- 10369196 TI - Management of IgA nephropathy: evidence-based recommendations. AB - The condition known as IgA nephropathy was first identified when Berger observed mesangial staining for IgA in healthy patients with isolated hematuria. These patients often presented with recurrent synpharyngitic hematuria or less frequently with asymptomatic microscopic hematuria and proteinuria. Although initially considered benign, we now recognize it as a common cause of end-stage renal failure. The overall prognosis may be better than suggested in the literature, as patients with mild asymptomatic hematuria are often not biopsied and, therefore, frequently are not included in published articles. We reviewed prospective and retrospective adult studies published after 1976 and analyzed them to produce evidence-based recommendations. Patients with proteinuria over 3 g/day, mild glomerular changes only, and preserved renal function (creatinine clearance over 70 ml/min) should be treated with prednisone. Steroids reduce proteinuria (grade B recommendation) and stabilize kidney function (grade C). The combination of cyclophosphamide, dipyridamole and warfarin should not be used (grade A), nor should cyclosporine A (grade B). In patients with progressive disease (creatinine clearance of less than 70 ml/min), fish oil should be given (grade B). A tonsillectomy could reduce proteinuria and hematuria in those patients with recurrent tonsillitis (grade D). Those with hypertension should be treated promptly with an angiotensin-converting enzyme inhibitor (grade B). PMID- 10369198 TI - Inhibins: paracrine and endocrine effects in female reproductive function. AB - A great deal of new information has arisen in the past 2 years concerning the physiology of inhibins and their clinical relevance in reproductive medicine. It is now recognized that the two inhibin isoforms, inhibin A and inhibin B, are produced by the gonads in the course of gamete maturation and have different patterns of secretion during the menstrual cycle. Inhibins are also produced by the placenta and fetal membranes and may be involved in physiological adaptation of pregnancy. Clinically, inhibins may serve as sensitive tumor markers in postmenopausal women, or as useful tools for evaluating ovarian reserve in infertile women; they may also be used in the diagnosis of materno-fetal disorders. PMID- 10369199 TI - The estrogen receptor family. AB - A significant flow of new data is currently being generated within the field of estrogen receptors and their mechanisms of action. This is primarily a result of the development of estrogen receptor knockout mice and the discovery of the second estrogen receptor, estrogen receptor beta. Both estrogen receptors appear to be involved in a multitude of regulatory events, the details of which will be worked out within the next few years. Estrogen receptor alpha appears to play a major role in the regulation of reproductive events and estrogen receptor alpha knockout female mice are completely infertile. Estrogen receptor beta knockout females have severe but incomplete infertility. Estrogen receptor beta gene mutations may, therefore, be of great clinical interest because they could perhaps explain some cases in which ovarian dysfunction leads to human infertility. Both receptors appear to be of essence for the cardiovascular system. Future studies will determine the relative importance of estrogen receptors alpha and beta in bone, the urogenital tract, the immune system, and the central nervous system, as well as in other estrogen target tissues. PMID- 10369200 TI - Vascular endothelial growth factor in reproductive biology. AB - The critical role of angiogenesis in embryology and tumor biology has been recognized for more than 20 years. However, the fact that neovascularization is essential to processes in mammalian female reproduction has only recently been appreciated widely. In this review we focus on a single angiogenic growth factor, vascular endothelial growth factor. As scientists have discovered in many aspects of cell biology, multiple and redundant signaling pathways have evolved in nature, presumably to protect essential biological functions from inactivating diseases or mutations. Despite this redundancy, some factors are of hierarchical importance. Vascular endothelial growth factor appears to be such a factor in the regulation of angiogenesis. PMID- 10369201 TI - Glycodelins: biological activity in reproduction and differentiation. AB - Studies published in recent years have elucidated the role of glycodelins in early human reproduction and the biological actions of glycodelins relative to their structure. New information has also emerged on the regulation of glycodelin synthesis and on the effects of glycodelin transfection on cellular growth and the expression of other epithelial markers. PMID- 10369203 TI - The metamorphosis of fertility management: lessons from assisted reproductive technology. PMID- 10369202 TI - Premenstrual syndromes: closing the 20th century chapters. AB - The need to re-evaluate premenstrual syndromes became apparent in 1997-1998 and early 1999. The success stories of some symptomatic treatment modalities and more sophisticated studies of pathobiology chart the pathways for future progress: the shift from a descriptive diagnosis to diagnoses based on etiology, the recognition of diversified vulnerabilities and their expression in particular situations, and specific treatment modalities. PMID- 10369204 TI - Advances in Y chromosome mapping. AB - The human Y chromosome has long been recognized as being responsible for sex determination. In fact, it also encodes more than 30 genes and gene families that participate in a variety of cellular functions, including bone development, tooth growth, and spermatogenesis. De-novo deletion of Y chromosome segments that contain spermatogenesis genes occurs frequently, resulting in low sperm production and male infertility. This article reviews our current knowledge of the structure and function of the Y chromosome is reviewed. PMID- 10369205 TI - Assisted reproduction for testicular failure: management of germ cell maturation arrest. AB - The availability of current, state of the art technology has enabled an efficient treatment in cases of infertility caused by testicular failure in which mature spermatozoa or late elongated spermatids can be recovered from the ejaculate or the testis. In those patients in whom only germ cells at earlier stages of spermatogenesis are present, results of the currently available treatments are still inconsistent and largely unpredictable, although some encouraging developments have been achieved over the past year. These new results suggest that developmental failure after fertilization with immature germ cells is mainly due to germ cell apoptosis and shows that an efficient selection of healthy germ cells can be achieved by in-vitro culture. In some cases, this method also allows premeiotic germ cells to achieve in-vitro transmeiotic maturation and thus enables men with in-vivo premeiotic maturation arrest to father a child. PMID- 10369206 TI - Immature oocyte retrieval with in-vitro oocyte maturation. AB - Early studies of in-vitro fertilization used immature oocytes. The process evolved to retrieving metaphase II oocytes, and was eventually successful. At present, aggressive ovulation induction protocols are the mainstay of assisted reproductive technology programs, but not without increased cost, multiple gestations, morbidity, potential future risks and isolated mortalities. The ability to retrieve each month's cohort of immature oocytes transvaginally opened the door to search for a new option for infertile couples requiring assisted reproductive technology. Immature oocyte retrieval combined with in-vitro oocyte maturation eliminates the stimulation, costs and time that were required to monitor oocytes, along with the short- and long-term complications. The essential components are optimal maturation media and a synchronized endometrium in which the embryos transferred from a truncated follicular phase can implant. The process has been successful in several centers with an acceptable success rate when used in conjunction with a host uterus. Future research with maturation, culture, and endometrial synchronization may allow immature oocyte retrieval with in-vitro oocyte maturation to replace in-vitro fertilization in its present form. PMID- 10369207 TI - Freezing of oocytes. AB - Storing oocytes is one of the challenges of assisted reproduction which may provide an alternative to embryo cryopreservation. Despite early disappointing results regarding survival, fertilization and cleavage rates, which led to only sporadic pregnancies in more than ten years, the recent introduction of technical modification greatly improved the clinical efficiency, with the birth of several healthy children. PMID- 10369208 TI - Screening oocytes and preimplantation embryos for aneuploidy. AB - Pregnancy and live birth rates following in-vitro fertilization decline rapidly with advancing maternal age partly because of an increase in age-related aneuploidies occurring in female meiosis. Screening oocytes or preimplantation embryos for common aneuploidies is now possible by polar body or cleavage stage biopsy and multicolour fluorescence in-situ hybridization. PMID- 10369209 TI - Culture and transfer of human blastocysts. AB - The transfer of the human embryo at the blastocyst stage during an in-vitro fertilization procedure is a way of increasing implantation rates. This, in turn, means that significantly fewer embryos are required to be transferred in order to establish a successful pregnancy. The result of this is that high order multiple gestations are eliminated, while maintaining high pregnancy rates, in in-vitro fertilization. PMID- 10369210 TI - Bibliography. Current world literature. Reproductive endocrinology. PMID- 10369211 TI - Bibliography. Current world literature. Fertility. PMID- 10369212 TI - H2O2-induced apoptotic death in serum-deprived cultures of oligodendroglia origin is linked to cell differentiation. AB - When deprived of serum, oligodendroglialike (OLN 93) cells grown on poly-L-lysine coated culture dishes cease to proliferate after 3 days and morphologically extend many fibers resembling morphologically differentiated, immature oligodendrocytes. At this time no cell death is apparent unless serum deprivation is extended for a period longer than 1 week. After 3 days in serum-deprived medium, treatment of cells with 1 mM H2O2 for 30 min facilitates apoptotic cell death, even when serum is added during the recovery period. Both serum-deprived, differentiated cells, and proliferating cells, respond to H2O2 by an initial growth arrest followed by growth resumption after 48 hr. However proliferating cells show resistance to the apoptotic effect of H2O2. This is correlated with growth arrest in the S phase at different stages of DNA replication, as well as with different timing of induced p21Waf1 expression. Thus, cells grown in serum, express elevated p21Waf1 protein levels after 4 hr, whereas serum-deprived, differentiated cells, only after 24 hr. The mRNA levels of p21Waf1 follow a similar timed pattern. Hence p21Waf1 may protect OLN 93 cells against the genotoxic effect of H2O2. The data suggest an intimate relationship between G1 arrest, morphological differentiation, and H2O2-mediated apoptosis. PMID- 10369213 TI - Superoxide mediates the cell-death-enhancing action of presenilin-1 mutations. AB - The mechanism whereby mutations in the presenilin-1 (PS-1) gene on chromosome 14 cause early-onset inherited Alzheimer's disease are unknown. We report that PC6 neural cells (a subclone of PC12 cells) expressing PS-1 mutations (M146V and L286V) exhibit increased superoxide production, nitrotyrosine accumulation, and membrane lipid peroxidation following exposure to amyloid beta-peptide 1-42 (Abeta). Mitochondrial calcium accumulation and membrane depolarization following exposure to Abeta were enhanced in cells expressing mutant PS-1. Overexpression of mitochondrial Mn-SOD greatly reduced superoxide production, nitrotyrosine formation, membrane lipid peroxidation, intramitochondrial calcium accumulation, and membrane depolarization following exposure to Abeta and conferred resistance to the apoptosis-enhancing action of the PS-1 mutations. Nitric oxide synthase inhibitors and the peroxynitrite scavenger uric acid blocked the apoptosis enhancing action of PS-1 mutations. The data suggest pivotal roles for superoxide production and resulting peroxynitrite formation in the pathogenic mechanism of PS-1 mutations. PMID- 10369214 TI - Estrogen and NGF synergistically protect terminally differentiated, ERalpha transfected PC12 cells from apoptosis. AB - The potential cytoprotective effects of the gonadal steroid estrogen, acting via its receptor (ER) on target neurons, are of considerable interest in aging, disease, and trauma. In this study, we examined the effects of estrogen on a prototypical neuronal-like cell, namely, nerve growth factor (NGF) differentiated PC12 cells when these cells are placed into an apoptosis-inducing environment. A clonal line of PC12 cells stably transfected to express full-length rat ERalpha mRNA and protein (PCER cells), as well as control cells (vector DNA transfected: PCCON cells), were differentiated into neurite bearing cells by 14 days of NGF treatment. Reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry for ERalpha revealed that terminally differentiated PCER cells continue to show robust ER mRNA and protein expression after differentiation. PCCON cells do not express ERalpha either before or after NGF induced differentiation. After differentiation, serum and NGF withdrawal from wild type PC12 cells is well known to induce cell death via apoptotic mechanisms. We challenged differentiated PCER and PCCON cells with serum-free media and assessed the effects of various treatments on cell survival and apoptosis using trypan blue staining and quantitative TUNEL staining. When differentiated PCER cells were treated with 17beta-estradiol (E2) plus NGF during a 2-day serum-free media challenge, cell survival was significantly greater, and the apoptotic index was significantly lower, than when PCER cells were treated with NGF only during the challenge. No additional improvement in cell survival, and no further reduction in apoptosis, was observed in non-ER expressing cells (PCCON) by E2 plus NGF treatment over that obtained by NGF alone. The effects of E2 on the expression of several apoptosis-related genes in this system were also examined. We found that E2 treatment of differentiated PCER cells in serum-free media increased the levels of Bcl-XL mRNA and reduced the levels of BAD mRNA relative to those in vehicle-treated PCER cells, and also relative to those in PCCON cells. Thus, estrogen's protective effects on PCER cells appear to, at least partly, involve a modulation of apoptosis-regulating genes. PMID- 10369215 TI - Localized expression of BMP and GDF mRNA in the rodent brain. AB - Expression of BMP- and GDF-related factors within the transforming growth factor beta (TGF-beta) superfamily was examined in the rat and mouse brain by in situ hybridization. Strong signals were obtained in neurons for GDF-1 and GDF-10. GDF 1 is expressed at postnatal day 6 in the cerebral cortex, hippocampal CA1 through CA3 neurons, while only weakly expressed by cells in the dentate gyrus. Granule cells and neurons in the polymorph layer of the dentate gyrus are GDF-1-positive, as are the majority of neurons in the cortex. GDF-10 shows a distinct pattern of expression: At P6, strong labelling was seen in the superficial layers of cortex, notably in the posterior cingulate cortex, and in CA3 and dentate gyrus. From postnatal day 21, GDF-1 expression is strong in the hippocampus, cortex, and thalamic nuclei, while GDF-10 expression becomes restricted to the granule cell layer in the dentate gyrus. In contrast, OP-1 expression is restricted throughout development to cells of the medial habenular nucleus, choroid plexus, and leptomeninges. The markedly different expression patterns of these BMPs suggest they serve separate functions in the brain. PMID- 10369216 TI - Glial depolarization evokes a larger potassium accumulation around oligodendrocytes than around astrocytes in gray matter of rat spinal cord slices. AB - The cell membrane of astrocytes and oligodendrocytes is almost exclusively permeable for K+. Depolarizing and hyperpolarizing voltage steps produce in oligodendrocytes, but not in astrocytes, decaying passive currents followed by large tail currents (Itail) after the offset of a voltage jump. The aim of the present study was to characterize the properties of Itail in astrocytes, oligodendrocytes, and their respective precursors in the gray matter of spinal cord slices. Studies were carried out on 5- to 11-day-old rats, using the whole cell patch clamp technique. The reversal potential (Vrev) of Itail evoked by membrane depolarization was significantly more positive in oligodendrocytes ( 31.7+/-2.58 mV, n = 53) than in astrocytes (-57.9+/-2.43 mV, n = 21), oligodendrocyte precursors (-41.2+/-3.44 mV, n = 36), or astrocyte precursors ( 52.1+/-1.32 mV, n = 43). Analysis of the Itail (using a variable amplitude and duration of the de- and hyperpolarizing prepulses as well as an analysis of the time constant of the membrane currents during voltage steps) showed that the Itail in oligodendrocytes arise from a larger shift of K+ across their membrane than in other cell types. As calculated from the Nernst equation, changes in Vrev revealed significantly larger accumulation of the extracellular K+ concentration ([K+]e) around oligodendrocytes than around astrocytes. The application of 50 mM K+ or hypotonic solution, used to study the effect of cell swelling on the changes in [K+]e evoked by a depolarizing prepulse, produced in astrocytes an increase in [K+]e of 201% and 239%, respectively. In oligodendrocytes, such increases (22% and 29%) were not found. We conclude that K+ tail currents, evoked by a larger accumulation of K+ in the vicinity of the oligodendrocyte membrane, could result from a smaller extracellular space (ECS) volume around oligodendrocytes than around astrocytes. Thus, in addition to the clearance of K+ from the ECS performed by astrocytes, the presence of the K+ tail currents in oligodendrocytes indicates that they might also contribute to efficient K+ homeostasis. PMID- 10369217 TI - Reinnervation of denervated lumbar ventral roots and their target muscle by thoracic spinal motoneurons via an implanted nerve autograft in adult rats after spinal cord injury. AB - Intraspinally implanting a nerve autograft (NAG) to promote axonal regeneration toward periphery was investigated as a surgical treatment for spinal cord injury in adult rats. Fifteen animals underwent a left hemisection of the spinal cord at T12 level and an intradural section of all ipsilateral lumbar ventral roots. In repaired animals (n = 9), the electrophysiologically selected left L3 and L4 lumbar ventral roots supplying the quadriceps muscle were anastomosed to a NAG. The NAG was taken from the right peroneal nerve and then ventrolaterally implanted into the cord at a level 7 mm rostral to the hemisection. In the control group (n = 6), sectioned lumbar ventral roots were left unrepaired. Nine months later, the animals were assessed with clinical, electrophysiological, and histological examinations. Muscle action potential and motor evoked potential were obtained from the denervated/reinnervated quadriceps in all repaired animals, with a mean amplitude of 918.3+/-328.9 microV and 215.8+/-39.7 microV, respectively. Horseradish peroxidase retrograde labeling from the denervated/repaired lumbar ventral roots, performed in five repaired animals, showed that the mean of labeled neurons, ipsilaterally located in the thoracic ventral horn near the implantation site, was 145.8+/-111.7. Histological analysis showed numerous myelinated axons in the NAG and denervated/repaired lumbar ventral roots of all repaired animals. The study of neuromuscular junctions furthermore confirmed numerous newly formed endplates appearing in the denervated/reinnervated quadriceps. These changes were absent in the control animals. These data indicate that the rostral thoracic spinal motoneurons can innervate the caudal denervated/repaired lumbar ventral roots and the target quadriceps via an implanted NAG, thereby inducing some functional recovery in adult rats after lower thoracic spinal cord injury. PMID- 10369218 TI - Pax-3 regulates neurogenesis in neural crest-derived precursor cells. AB - The peripheral nervous system consists of multiple neural lineages derived from the neural crest (NC). Pax-3 is expressed in the NC and when mutated in the splotch mouse (Sp) results in the loss of derivatives from this precursor cell population. We have investigated the role of Pax-3 in regulating the generation of neurons from NC-derived precursor cells in vitro. Pax-3 mRNA in NC cultures is initially expressed in all NC but is subsequently only retained in neurons, suggesting a role in their generation. To determine whether Pax-3 is involved in neuron development, we first examined the generation of sensory-like neurons in NC cultures from Sp mice. Fivefold less sensory-like neurons were generated in NC cultures from Sp homozygous mice as compared to wild-type littermates. The role of Pax-3 in sensory neuron generation was then directly examined in dorsal root ganglia cultures by down-regulating the expression of Pax-3 protein with antisense oligonucleotides. It was found that antisense oligonucleotides inhibited 80-90% of newly generated sensory neurons; however, there was no significant effect on the survival of sensory neurons or the precursor population. These results suggest that Pax-3 has a role in regulating the differentiation of peripheral neurons. PMID- 10369219 TI - Developmental regulation of the serotonergic transmitter phenotype in rostral and caudal raphe neurons by transforming growth factor-betas. AB - Serotonergic (5-HT) neurons of the CNS develop as two separate clusters, a rostral and a caudal group, within the brain stem raphe. We show here that the transforming growth factors -beta2 and -beta3 (TGF-beta) and the TGF-beta type II receptor are expressed in the embryonic rat raphe, when 5-HT neurons develop and differentiate. To investigate putative roles of TGF-betas in the regulation of 5 HT neuron development we have generated serum-free cultures isolated either from the rostral or the caudal embryonic rat raphe, respectively. In cultures from the caudal E14 raphe saturating concentrations (5 ng/ml) of TGF-beta2 and -beta3 augmented numbers of tryptophan hydroxylase (TpOH) -immunoreactive neurons and cells specifically taking up 5,7-dihydroxytryptamine (5,7-DHT) by about 1.7-fold over a period of 4 days. Treatment with TGF-betas also increased uptake of 3H-5HT uptake about 1.7-fold. Alterations in 5-HT neuron numbers were due to the induction of serotonergic markers rather than increased survival, as shown by the efficacy of delayed short-term treatments. Comparing rostral and caudal raphe cultures from different embryonic ages suggests that distinct effects of TGF betas reflect the responsiveness of 5-HT neurons at different ages rather than of different origins. PMID- 10369220 TI - SNAP-25 requirement for dendritic growth of hippocampal neurons. AB - Structure and dimension of the dendritic arbor are important determinants of information processing by the nerve cell, but mechanisms and molecules involved in dendritic growth are essentially unknown. We investigated early mechanisms of dendritic growth using mouse fetal hippocampal neurons in primary culture, which form processes during the first week in vitro. We detected a key component of regulated exocytosis, SNAP-25 (synaptosomal associated protein of 25 kDa), in axons and axonal terminals as well as in dendrites identified by the occurrence of the dendritic markers transferrin receptor and MAP2. Selective inactivation of SNAP-25 by botulinum neurotoxin A (BoNTA) resulted in inhibition of axonal growth and of vesicle recycling in axonal terminals. In addition, dendritic growth of hippocampal pyramidal and granule neurons was significantly inhibited by BoNTA. In contrast, cleavage of synaptobrevin by tetanus toxin had an effect on neither axonal nor dendritic growth. Our observations indicate that SNAP-25, but not synaptobrevin, is involved in constitutive axonal growth and dendrite formation by hippocampal neurons. PMID- 10369221 TI - Potentiation of GABA(A) receptor-mediated Cl-current by urotensin peptides in identified Aplysia neurons. AB - Gamma-aminobutyric acid (GABA) is the main inhibitory neurotransmitter in the vertebrate and invertebrate central nervous systems, including those of molluscs. The effects of extracellularly applied urotensin peptides (urotensin I (UI) and urotensin II (UII)) on the GABA-induced Cl- current recorded from identified neurons (R9 and R12) of Aplysia kurodai were investigated using voltage-clamp and pressure ejection techniques. Focal application of 100 nM UI and UII potentiated the GABA-induced Cl- current without affecting the resting membrane conductance and holding current. The increase was completely reversible. The GABA-induced Cl- current also was potentiated by bath-applied UI and UII (5-10 nM). The potentiating effects of UI and UII on the GABA-induced Cl- current were concentration-dependent and completely reversible. These results suggest that neurotensin peptides may decrease neuronal excitability by potentiating the GABA(A) receptor-mediated Cl- current in the neurons of mammalian and invertebrate central nervous systems. PMID- 10369222 TI - Differential, regional, and cellular expression of the stathmin family transcripts in the adult rat brain. AB - Stathmin is a ubiquitous cytosolic phosphoprotein, preferentially expressed in the nervous system, and previously described as a relay integrating diverse intracellular signaling pathways. Stathmin is the generic element of a mammalian protein family including SCG10, SCLIP, and RB3 with its splice variants RB3' and RB3". In contrast with stathmin, SCG10, SCLIP, and RB3/RB3'/RB3" are exclusively expressed in the nervous system, stathmin and SCG10 being mostly expressed during cell proliferation and differentiation, and SCLIP and RB3 rather in mature neural cells. To further understand their specific roles in the CNS, we compared the localization of the stathmin, SCG10, SCLIP, and RB3 transcripts in adult rat brain. Northern blot analysis as well as in situ hybridization experiments showed that all stathmin-related mRNAs are expressed in a wide range of adult rat brain areas. At a regional level, SCG10 and SCLIP appear generally distributed similarly except in a few areas. The pattern of expression of the RB3 transcript is very different from that of the three other members of the stathmin family. Furthermore, unlike SCG10 and SCLIP, which were detected only in neurons, but like stathmin, RB3 was detected in neurons and also in glial cells of the white matter. Altogether, our results suggest distinct roles for each member of the stathmin-related phosphoprotein family, in regard to their specific regional and cellular localization in the rat brain. PMID- 10369223 TI - What are physicians thinking? A survey of first-year medical students. PMID- 10369224 TI - Delayed post-traumatic cervical instability. AB - BACKGROUND: Cervical spine instability is a clinical entity whose biomechanical and radiological features have been widely discussed by many authors. On the other hand, the subject of delayed post-traumatic cervical instability is often surrounded by confusion due to its difficult nosologic framing; the aim of this study is to contribute to the matter. METHODS: A cooperative study was organized by the Study Group for Spinal Surgery of the Italian Society of Neurosurgery to evaluate cervical trauma patients surgically treated more than 20 days after the traumatic event. From a total number of 172 patients, twenty-five were admitted to the study, because neuroradiological investigations performed during the acute phase had shown either an absence of traumatic lesions or only minimal lesions judged to be stable. For this reason these 25 patients had not been treated by either surgery or immobilization in a halo vest. Some time after trauma, this group of patients clearly demonstrated evidence of unstable lesions requiring surgical treatment, following the appearance of new clinical signs or on neuroradiological follow-up. RESULTS: Re-examination of the neuroradiological investigations performed during the acute phase made it possible to identify elements that might have led us to suspect the presence of ligamental lesions: microfractures, dislocations less than 3 mm, and inversion of physiological lordosis. CONCLUSIONS: This review clearly indicates that patients with even mild cervical trauma must be scrupulously evaluated during the acute phase and that in some cases it is advisable to perform a more detailed neuroradiological investigation. PMID- 10369225 TI - Posterior C1-C2 transarticular screw fixation in the treatment of displaced type II odontoid fractures in the geriatric population--review of seven cases. AB - BACKGROUND: Seven geriatric patients presented with displaced Type II odontoid fractures. All patients underwent a posterior C1-C2 transarticular fixation between November 1994 and December 1996. Ages ranged between 63 and 88 years. METHODS: Fractures were treated with placement of bilateral transarticular screws, allowing immediate fixation, except in one patient, for whom only a unilateral screw was used. An autograft interspinous strut was also placed, allowing three-point fixation. Mean follow-up was 10.6 months. RESULTS: Six patients received rigid fixation and developed a stable union. One patient died before any follow-up could be obtained. Two other patients died within 1 year of unrelated causes. The remaining four patients remain active and independent. One intraoperative vertebral artery injury was identified. No clinical sequalae were noted. CONCLUSION: Posterior transarticular screw fixation is a reasonable option in treating these controversial fractures. Seven geriatric patients tolerated this surgery well, and were mobilized early, avoiding complications related to external immobilization. PMID- 10369226 TI - Atypical forms of spinal tuberculosis: case report and review of the literature. AB - OBJECTIVE: The object of this report is to highlight some of the less known atypical features of spinal tuberculosis (TB) in the hope of facilitating early diagnosis. Pure neural arch and sacral TB is rare and the co-existence of these two as widely separated skip lesions in the same patient is even rarer. CLINICAL PRESENTATION: An unusual case of tuberculous process affecting the sacrum as well as the neural arches of upper cervical vertebrae is presented. Neither the clinical features nor the imaging techniques, including radiography, bone scintigraphy, computed tomography, and magnetic resonance imaging, were helpful in establishing the diagnosis. The destructive lesion of the sacrum with a rectally palpable presacral mass was thought to be a chordoma or chondrosarcoma until the patient developed upper cervical cord compression with an extradural myelographic block. Development of this second destructive lesion involving the posterior spinal elements (the neural arch) led to a diagnosis of malignant spinal metastasis. The true diagnosis was only revealed by the histology of the solid tumor-like extradural mass in the upper cervical region and demonstration of acid-fast bacilli (AFB) in the lesion. Anti-TB chemotherapy resulted in complete resolution of sacral and cervical lesions as well as the neurologic deficits. CONCLUSION: Differential diagnosis of the obscure spinal lesion should include tuberculosis, specifically the atypical forms; especially because complete cure is possible with early treatment and neurologic morbidity is high in neglected cases. PMID- 10369227 TI - Ruptured anterior spinal artery aneurysm: a case report. AB - BACKGROUND: Spinal artery aneurysms are rare, and are usually found in association with arteriovenous malformations or coarctation of the aorta. CASE REPORT: A 42-year-old man with a ruptured anterior spinal artery aneurysm is presented here. He experienced subarachnoid hemorrhage, which was confirmed by computed tomography. Magnetic resonance imaging revealed an aneurysm in front of the upper part of the medulla. Angiography demonstrated bilateral vertebral artery occlusion. Distal vertebral arteries and the basilar artery were perfused via the dilated anterior spinal artery, which originates in the right subclavian artery. The aneurysm was located at the distal part of the anterior spinal artery, and was successfully clipped through a lateral suboccipital craniotomy 2 months after bleeding from the aneurysm. After rehabilitation, the patient was able to walk with no apparent neurologic deficit. CONCLUSIONS: This case suggests that the anterior spinal artery as a collateral route after bilateral vertebral artery occlusion is under hemodynamic stress, resulting in aneurysm formation and rupture. PMID- 10369228 TI - Post-traumatic brain stem distortion: a case report. AB - BACKGROUND: Primary traumatic brain stem injury (BSI) lesions are found most frequently in the dorsal or dorsolateral midbrain, whereas distortion of the brain stem itself is exceedingly rare. CASE REPORT: We present a 20-year-old woman with a rare brain stem injury caused by a violent motor vehicle collision. Magnetic resonance imaging at 2 months after injury revealed marked brain stem distortion with loss of the normal shape at the midbrain and pons, which were displaced anteriorly in association with the fracture of the clivus. Moreover, the medulla oblongata showed a loose winding configuration. At discharge about 5 months after injury, the Glasgow Outcome Scale was severe disability. CONCLUSION: This BSI was caused by reciprocal actions of fracture of the clivus and the direct effect on the brain stem caused by acceleration or rotational forces. PMID- 10369229 TI - Penetrating intracranial wooden object: case report and review of CT morphology, complications, and management. AB - BACKGROUND: Penetrating intracranial wooden fragments after vehicular accidents are uncommon. The CT morphology, complications, and management in such cases are quite variable. CASE REPORT: A 27-year-old male was seen with a "twig" from a tree embedded firmly just below the right medial canthus after a motorcycle accident. Diagnosis of intracranial penetrating wooden object was made on CT scanning. The wooden stick, which had splintered into two, was extricated through a craniotomy in two operative sessions. However the patient succumbed to septicemia and meningitis on the twelfth day after the accident. CONCLUSIONS: The need for prompt extrication of these objects and the causes of high mortality in this condition are discussed. The importance of imaging the intracranial compartment in injuries involving the periorbital region is emphasized. PMID- 10369230 TI - Choroid plexus papilloma of the cerebellopontine angle: a twelve patient series. AB - BACKGROUND: Choroid plexus papillomas (CPPs), of the cerebellopontine angle (CPA), are a rare entity and no surgical series have been reported so far. We reviewed all the pertinent literature of 12 patients operated on in the last 20 years at our institution. METHODS: All the patients were adults, ranging from 19 to 61 years. The group included 6 males and 6 females. Preoperatively, on computerized tomography (CT) (n = 10) or magnetic resonance imaging (MRI) (n = 4), differential diagnosis was difficult to achieve, especially with meningiomas. Hydrocephalus was disclosed in 8 cases. Angiography (n = 11) showed tumor blush with typical vascular supply in almost half the cases. RESULTS: In 6 patients a midline approach via the cerebellomedullary fissure was used; in the remaining 6 patients the retromastoid route was adopted. We found 2 "unconnected" tumors; no hilum was identified at surgery. Total tumor removal was achieved in 6 patients, predominantly in the last cases. Two patients died in the postoperative period, 3 patients had mild additional deficits, whereas 7 patients were stable or improved. All the patients were followed up for a mean period of 8.2 years. Conventional radiotherapy was carried out in 5 patients; 1 of them after tumor recurrence. Stereotactic radiotherapy was performed in 3 patients; 2 of them after recurrences. Two patients showed tumor progression and died during the follow-up. One of them presented a suprasellar metastasis and died much earlier (2 versus 7 years). CONCLUSION: Careful assessment and surgical planning is accomplished with the combined information from CT, MRI, and angiography. Typical characteristics are the following: vascular supply from the choroidal arteries, ventral extension, adhesion to the brainstem, and lower cranial nerves. Nowadays, total removal of CPPs of the CPA can be achieved with acceptable morbidity and mortality. In our experience, conventional radiotherapy did not prove effective. PMID- 10369231 TI - Extraventricular cerebral anaplastic ependymomas. AB - BACKGROUND: Anaplastic ependymomas are considered to be uncommon cerebral tumors by most authors. We have had the opportunity to study 34 cases of such lesions in 13 years. METHODS: 34 cases of anaplastic ependymoma operated in different hospitals of Maracaibo, Venezuela, during the period of 1983-1995 were analyzed. Cases of ependymoblastoma were excluded. RESULTS: Adult patients made up most of the present series. All patients harbored supratentorial growths in locations distant from the ventricular system. The microscopic pattern was of limited value to establish prognosis, for there was no constant correlation between the histologic features and tumor relapse; only in sporadic cases in which high cell density and conspicuous mitotic activity were maximally expressed, did tumor relapse occur shortly after removal of the lesion. CONCLUSION: This type of paradoxical behavior being the rule makes all attempts at predicting prognosis in these entities a disappointing task. PMID- 10369232 TI - Solitary fibrous tumor of the meninges: two new cases and review of the literature. AB - BACKGROUND: Solitary fibrous tumor (SFT), a mesenchymal neoplasm originally described in the pleura has been more recently reported to arise in a number of other sites, including the meninges. Nowadays immunohistochemistry facilitates the otherwise problematic differential diagnosis with regard to other benign and malignant spindle cell neoplasms of the central nervous system. METHODS: Two recently treated cases of meningeal SFT (one craniospinal, one spinal) are presented and discussed in the light of the present knowledge and a review of the literature. RESULTS: Total resection was followed by complete recovery and both patients are presently asymptomatic and without evidence of disease. The microscopic and immunohistochemical profiles (CD 34, vimentin positive; S-100, EMA negative) were consistent with those of previously reported cases. CONCLUSIONS: The majority of SFTs behave in a benign fashion and do not recur unless subtotally resected. Malignant variants may account for up to 37% of SFTs in other locations but have never been reported to occur in the meninges. Meningeal SFTs are to be considered a new pathological entity. Wider use of immunohistochemical screening should enable the determination of their real incidence; larger series and longer follow-up will provide conclusions about their treatment and prognosis. PMID- 10369233 TI - Endoscopic procedures for resection of arteriovenous malformations. AB - BACKGROUND: Resection of arteriovenous malformations (AVMs), particularly those located in functional areas, requires precision. To enhance that precision, endoscope-assisted microsurgery has been employed at Loma Linda University. METHODS: Twenty-five consecutive cases of AVM were treated microsurgically with endoscopic assistance. Patients were divided into two groups: (1) those having AVMs in functional areas, and (2) those whose AVMs extended into the ventricle, either in the trigonal area or the capsulocaudatothalamic area. The endoscope was inserted into the subarachnoid space to interrupt communicating venules around the major draining vein and into the cleavage developed between the AVM venous loops and surrounding brain tissue as shunting arterioles and communicating venules were interrupted. For surgery of intraventricular AVMs, the curved endoscope was inserted into the ventricle, providing visualization of the AVM core, which was dissected from the ventricular side. RESULTS: AVMs were totally resected in all cases except for two patients with capsulocaudatothalamic AVMs, which were decreased in size sufficiently to receive radiosurgery. CONCLUSION: Endoscope-assisted microsurgery enhances magnification, illumination, and technical precision while the surgeon is dissecting the AVM core vessels and while operating on AVMs extending into the ventricle. PMID- 10369234 TI - Is there a potential role for hyoid bone compression in pathogenesis of carotid artery stenosis? AB - BACKGROUND: Blood flow turbulence and increased shear stress, particularly at sites of sudden, marked arterial wall changes, are significant hemodynamic parameters in the pathogenesis of atherosclerosis. We present a case in which we found the hyoid bone protruding into the carotid vessels and may have been contributing, in part, to atherosclerotic carotid stenosis. CASE PRESENTATION: An 85-year-old woman presenting with left arm and leg weakness consistent with right hemispheric transient ischemic attack. Magnetic resonance arteriography (MRA) and carotid non-invasive studies revealed a 90% stenosis of the right internal carotid artery. At surgery, the hyoid bone on the right side was projecting into the internal carotid artery, causing indentation. There was associated rotation of the internal and external carotid arteries from their normal position. Right carotid endarterectomy was performed and the lateral one-third of the hyoid bone excised to alleviate the external compression. Postoperative spiral computerized tomography (CT) scan of the carotid vessels demonstrated the extent of hyoid resection as well as rotation of the external and internal carotid arteries. CONCLUSIONS: We suggest the possible contribution of hyoid bone compression to the pathogenesis of atherosclerotic carotid artery stenosis. This report also highlights the diagnostic value of CT angiography in the assessment of carotid artery occlusive disease. PMID- 10369235 TI - Measurement of ischemia by changes in tissue oxygen, carbon dioxide, and pH. AB - BACKGROUND: We evaluated the ability of brain tissue oxygen pressure (PO2), carbon dioxide pressure (PCO2), and pH to detect regional ischemia produced by temporary brain artery occlusion, compared with a group without artery occlusion. METHODS: Patients undergoing craniotomy for cerebrovascular surgery were recruited for this study. A 0.5-mm-diameter probe was inserted into brain tissue to measure PO2, PCO2, and pH continuously. Group 1 (n = 15) did not receive brain artery occlusion during their surgical procedure. In Group 2, brain artery occlusion was produced for aneurysm clipping (n = 10) or extracerebral to intracerebral artery bypass (n = 3). Mean arterial pressure was maintained above 90 mmHg in both groups. Measurements were made after artery occlusion or sham treatment and compared with baseline. RESULTS: Under baseline conditions, tissue PO2, PCO2, and pH were not different between the groups. In Group 2, brain artery occlusion for a median time of 7 minutes (range, 2-48 min) significantly decreased PO2 and pH and increased PCO2 compared with baseline. There were no significant changes in Group 1. During artery occlusion, PO2 decreased below 10 mmHg and/or pH decreased below 7.0 in 8 of 13 patients. CONCLUSIONS: Regional brain ischemia can be consistently detected and treated by monitoring tissue metabolism. It will be necessary in the future to identify critical levels and duration of decreases in PO2 and pH that lead to irreversible neuronal injury. PMID- 10369236 TI - Revision and removal of stimulating electrodes following long-term therapy with the vagus nerve stimulator. AB - BACKGROUND: A significant concern about vagus nerve stimulation therapy has been the disposition of the spiral stimulating electrodes once treatment is considered ineffective or is no longer desired. Because the electrodes are wrapped around the vagus nerve, there is the potential for nerve injury during their removal. METHODS: We attempted removal of the spiral stimulating electrodes from 10 patients who received long-term vagus nerve stimulation therapy for drug resistant epilepsy. In some patients, replacement with electrodes was also performed for poorly functioning leads. RESULTS: The mean duration of electrode implantation was 3.7+/-2.2 years (range 1.1-7.3 years). In seven patients, the old electrodes were removed completely from the nerve. No adverse events occurred intraoperatively or postoperatively. CONCLUSIONS: Our results indicate that the spiral electrodes may be safely removed from the vagus nerve, even after the electrodes have been implanted for several years. The reversibility of lead implantation may enhance the attractiveness of vagus nerve stimulation therapy for patients with medically-intractable epilepsy. PMID- 10369237 TI - Computer analysis of the tonic, phasic, and kinesthetic activity of pallidal discharges in Parkinson patients. AB - OBJECTIVE: Intraoperative analysis of microrecording data during pallidotomy often depends on subjective interpretation of the oscilloscope signal, especially during the analysis of phasic activity. The goals of this project were: 1) to develop an inexpensive system that allowed on-line, objective characterization of single-unit pallidal discharges, and 2) to have objective criteria to differentiate the internal part (GPi) from the external part (GPe) of the globus pallidus. METHODS: A computer program was developed that allowed the analysis of firing rates (mean, median, and quartiles), spike count per unit time, and interspike interval (ISI) histograms with Chi-square statistical evaluation. Indices were developed that measured phasic activity, including burst index (BI) for the measurement of bursts, pause index (PI) for the measurement of pauses, and pause ratio (PR) for analysis of time spent in pauses. Single-unit activity of 152 GPe and 203 GPi cells in 47 Parkinson patients were digitized using the computer soundcard during pallidotomy and analyzed using this software. RESULTS: GPe discharges had a mean firing rate = 42 Hz, BI = 0.81, PI = 0.21, and PR = 1.41. GPi had a mean firing rate = 81, BI = 1.61, PI = 0.04, and PR = 0.21. The PR was the best index that differentiated GPe from GPi, followed by PI, BI, and firing rates, in that order. Kinesthetic cells were recorded equally in GPe from GPi, and their responses to generalized movements were not significantly different. CONCLUSION: (1) Signal analysis using the digitization process of a computer sound card and dedicated software is satisfactory for the objective "on line" and "off-line" analysis of microrecordings (including phasic activity); (2) PI and PR are most helpful in differentiating neurons of GPi from those of GPe; (3) no single parameter can differentiate GPe from GPi activity in all cases; and (4) unlike the findings in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) treated monkeys, GPe and GPi of Parkinson patients have similar prevalence of kinesthetic cells and similar responses to generalized somatotopic effects. PMID- 10369238 TI - Direct identification of ventrointermediate nucleus of the thalamus on magnetic resonance and computed tomography images. AB - OBJECTIVE: The ventro-intermediate (Vim) nucleus of the thalamus is a commonly used target for the treatment of tremor. The thalamic fasciculus contains myelinated fibers, believed to play a role in the generation of tremor, that converge into a dense bundle at the inferior aspect of the Vim nucleus, making it visible on magnetic resonance (MR) and computed tomography (CT) images. This structure, therefore, can be visualized directly and targeted for thalamotomy. METHODS: Thalamotomies were performed on nine patients (who have a follow-up of 13-23 months) with parkinsonian and essential tremors using MR and CT images. The tremor target was hypointense on MR images obtained in inversion recovery sequence and hypointense on CT images. It was therefore visualized, directly targeted, and probed. Stimulation studies were done to physiologically confirm accuracy of the probe placement and then a radiofrequency lesion was made. RESULTS: Stimulation of the target identified as the Vim nucleus on MR and CT images produced responses similar to those expected from the Vim nucleus. After this site was lesioned tremor disappeared in all nine patients. CONCLUSION: The Vim nucleus of the thalamus is visible on MR and CT images. Destruction of this target abolishes parkinsonian and essential tremors. PMID- 10369239 TI - An interview with Professor Hajime Handa. PMID- 10369240 TI - Neurosurgical heritage. Fathers/sons/daughters in neurosurgery in the U.S. and Canada. PMID- 10369241 TI - Brain stem epidermoid cyst. PMID- 10369242 TI - Arachnoid cyst and epilepsy. PMID- 10369243 TI - Breaking waves down under. PMID- 10369244 TI - Chips for chimps. PMID- 10369245 TI - Creating deaths from malaria. PMID- 10369246 TI - You give me fever. PMID- 10369247 TI - A mutation in the gene encoding steroidogenic factor-1 causes XY sex reversal and adrenal failure in humans. PMID- 10369248 TI - Cloning of male mice from adult tail-tip cells. PMID- 10369250 TI - An estimated frequency of endogenous insertional mutations in humans. PMID- 10369249 TI - Detection of a cystic fibrosis modifier locus for meconium ileus on human chromosome 19q13. PMID- 10369251 TI - Imprinting and monogamy. PMID- 10369252 TI - Genetic conflicts and the private life of Peromyscus polionotus. PMID- 10369253 TI - The future of genetic counselling: an international perspective. AB - The focus of clinical genetics, and thus genetic counselling, is forecast to expand from the diagnosis and prediction of rare, often untreatable conditions, to the prediction of common, often treatable or preventable conditions. Whether this evolution is likely to proceed rapidly or at a pace that permits sensible integration of molecular genetic tools is unknown and a source of debate. It is clear, however, that genetic discoveries will modify the way in which disease and risk are conceptualized. Here, we predict how genetic counselling, specifically for more common diseases, might be provided in the decades to come. We envisage an expansion of professional roles and expertise for many health care providers and highlight the need for counselling practices to become more evidence based. Although we support an evidentiary-based approach to the integration of genetic testing into practice, genetic advance is unlikely to occur in an orderly and standardized manner within countries, much less among different countries and health care systems. Geneticists will become increasingly involved in professional education and policy-making regarding genetic testing and screening programs. PMID- 10369254 TI - Prospects for whole-genome linkage disequilibrium mapping of common disease genes. AB - Recently, attention has focused on the use of whole-genome linkage disequilibrium (LD) studies to map common disease genes. Such studies would employ a dense map of single nucleotide polymorphisms (SNPs) to detect association between a marker and disease. Construction of SNP maps is currently underway. An essential issue yet to be settled is the required marker density of such maps. Here, I use population simulations to estimate the extent of LD surrounding common gene variants in the general human population as well as in isolated populations. Two main conclusions emerge from these investigations. First, a useful level of LD is unlikely to extend beyond an average distance of roughly 3 kb in the general population, which implies that approximately 500,000 SNPs will be required for whole-genome studies. Second, the extent of LD is similar in isolated populations unless the founding bottleneck is very narrow or the frequency of the variant is low (<5%). PMID- 10369255 TI - A polymorphism that affects OCT-1 binding to the TNF promoter region is associated with severe malaria. AB - Genetic variation in cytokine promoter regions is postulated to influence susceptibility to infection, but the molecular mechanisms by which such polymorphisms might affect gene regulation are unknown. Through systematic DNA footprinting of the TNF (encoding tumour necrosis factor, TNF) promoter region, we have identified a single nucleotide polymorphism (SNP) that causes the helix turn-helix transcription factor OCT-1 to bind to a novel region of complex protein-DNA interactions and alters gene expression in human monocytes. The OCT-1 binding genotype, found in approximately 5% of Africans, is associated with fourfold increased susceptibility to cerebral malaria in large case-control studies of West African and East African populations, after correction for other known TNF polymorphisms and linked HLA alleles. PMID- 10369256 TI - Hyperornithinaemia-hyperammonaemia-homocitrullinuria syndrome is caused by mutations in a gene encoding a mitochondrial ornithine transporter. AB - Neurospora crassa ARG13 and Saccharomyces cerevisiae ARG11 encode mitochondrial carrier family (MCF) proteins that transport ornithine across the mitochondrial inner membrane. We used their sequences to identify EST candidates that partially encode orthologous mammalian transporters. We thereby identified such a gene (ORNT1) that maps to 13q14 and whose expression, similar to that of other urea cycle (UC) components, was high in liver and varied with changes in dietary protein. ORNT1 expression restores ornithine metabolism in fibroblasts from patients with hyperammonaemia-hyperornithinaemia-homocitrullinuria (HHH) syndrome. In a survey of 11 HHH probands, we identified 3 ORNT1 mutant alleles that account for 21 of 22 possible mutant ORNT1 genes in our patients: F188delta, which is common in French-Canadian HHH patients and encodes an unstable protein; E180K, which encodes a stable, properly targeted protein that is inactive; and a 13q14 microdeletion. Our results show that ORNT1 encodes the mitochondrial ornithine transporter involved in UC function and is defective in HHH syndrome. PMID- 10369257 TI - The gene mutated in adult-onset type II citrullinaemia encodes a putative mitochondrial carrier protein. AB - Citrullinaemia (CTLN) is an autosomal recessive disease caused by deficiency of argininosuccinate synthetase (ASS). Adult-onset type II citrullinaemia (CTLN2) is characterized by a liver-specific ASS deficiency with no abnormalities in hepatic ASS mRNA or the gene ASS (refs 1-17). CTLN2 patients (1/100,000 in Japan) suffer from a disturbance of consciousness and coma, and most die with cerebral edema within a few years of onset. CTLN2 differs from classical citrullinaemia (CTLN1, OMIM 215700) in that CTLN1 is neonatal or infantile in onset, with ASS enzyme defects (in all tissues) arising due to mutations in ASS on chromosome 9q34 (refs 18-21). We collected 118 CTLN2 families, and localized the CTLN2 locus to chromosome 7q21.3 by homozygosity mapping analysis of individuals from 18 consanguineous unions. Using positional cloning we identified a novel gene, SLC25A13, and found five different DNA sequence alterations that account for mutations in all consanguineous patients examined. SLC25A13 encodes a 3.4-kb transcript expressed most abundantly in liver. The protein encoded by SLC25A13, named citrin, is bipartite in structure, containing a mitochondrial carrier motif and four EF-hand domains, suggesting it is a calcium-dependent mitochondrial solute transporter with a role in urea cycle function. PMID- 10369258 TI - Determination of ancestral alleles for human single-nucleotide polymorphisms using high-density oligonucleotide arrays. AB - Here we report the application of high-density oligonucleotide array (DNA chip) based analysis to determine the distant history of single nucleotide polymorphisms (SNPs) in current human populations. We analysed orthologues for 397 human SNP sites (identified in CEPH pedigrees from Amish, Venezuelan and Utah populations) from 23 common chimpanzee, 19 pygmy chimpanzee and 11 gorilla genomic DNA samples. From this data we determined 214 proposed ancestral alleles (the sequence found in the last common ancestor of humans and chimpanzees). In a diverse human population set, we found that SNP alleles with higher frequencies were more likely to be ancestral than less frequently occurring alleles. There were, however, exceptions. We also found three shared human/pygmy chimpanzee polymorphisms, all involving CpG dinucleotides, and two shared human/gorilla polymorphisms, one involving a CpG dinucleotide. We demonstrate that microarray based assays allow rapid comparative sequence analysis of intra- and interspecies genetic variation. PMID- 10369259 TI - Pharmacogenetic association between ALOX5 promoter genotype and the response to anti-asthma treatment. AB - Clinically similar asthma patients may develop airway obstruction by different mechanisms. Asthma treatments that specifically interfere with the 5-lipoxygenase (ALOX5) pathway provide a method to identify those patients in whom the products of the ALOX5 pathway (that is, the leukotrienes) contribute to the expression of the asthma phenotype. Failure of an asthma patient to respond to treatment with ALOX5-pathway modifiers indicates that leukotrienes are not critical to the expression of the asthmatic phenotype in that patient. We previously defined a family of DNA sequence variants in the core promoter of the gene ALOX5 (on chromosome 10q11.2) associated with diminished promoter-reporter activity in tissue culture. Because expression of ALOX5 is in part transcriptionally regulated, we reasoned that patients with these sequence variants may have diminished gene transcription, and therefore decreased ALOX5 product production and a diminished clinical response to treatment with a drug targeting this pathway. Such an effect indicates an interaction between gene promoter sequence variants and drug-treatment responses, that is, a pharmacogenetic effect of a promoter sequence on treatment responses. PMID- 10369260 TI - A double-stranded RNA binding protein required for activation of repressed messages in mammalian germ cells. AB - Chromatin packaging in mammalian spermatozoa requires an ordered replacement of the somatic histones by two classes of spermatid-specific basic proteins, the transition proteins and the protamines. Temporal expression of transition proteins and protamines during spermatid differentiation is under translational control, and premature translation of protamine 1 leads to precocious nuclear condensation and sterility. We have previously suggested that the double-stranded (ds) RNA binding protein Prbp (encoded by the gene Tarbp2) functions as a translational regulator during mouse spermatogenesis. Here we show that Prbp is required for proper translational activation of the mRNAs encoding the protamines. We generated mice that carry a targeted disruption of Tarbp2 and determined that they were sterile and severely oligospermic. Using immunohistological analysis, we determined that the endogenous Prm2 mRNA and a reporter mRNA carrying protamine 1 translational-control elements were translated in a mosaic pattern. We showed that failure to synthesize the protamines resulted in delayed replacement of the transition proteins and subsequent failure of spermiation. The timing of Prbp expression suggests that it may function as a chaperone in the assembly of specific translationally regulated ribonucleoprotein particles. PMID- 10369261 TI - Mutations in MVK, encoding mevalonate kinase, cause hyperimmunoglobulinaemia D and periodic fever syndrome. AB - Hyperimmunoglobulinaemia D and periodic fever syndrome (HIDS; MIM 260920) is an autosomal recessive disorder characterized by recurrent episodes of fever associated with lymphadenopathy, arthralgia, gastrointestinal dismay and skin rash. Diagnostic hallmark of HIDS is a constitutively elevated level of serum immunoglobulin D (IgD), although patients have been reported with normal IgD levels. To determine the underlying defect in HIDS, we analysed urine of several patients and discovered increased concentrations of mevalonic acid during severe episodes of fever, but not between crises. Subsequent analysis of cells from four unrelated HIDS patients revealed reduced activities of mevalonate kinase (MK; encoded by the gene MVK), a key enzyme of isoprenoid biosynthesis. Sequence analysis of MVK cDNA from the patients identified three different mutations, one of which was common to all patients. Expression of the mutant cDNAs in Escherichia coli showed that all three mutations affect the activity of the encoded proteins. Moreover, immunoblot analysis demonstrated a deficiency of MK protein in patient fibroblasts, indicating a protein-destabilizing effect of the mutations. PMID- 10369262 TI - Mutations in the gene encoding mevalonate kinase cause hyper-IgD and periodic fever syndrome. International Hyper-IgD Study Group. AB - Hyperimmunoglobulinaemia D and periodic fever syndrome (HIDS; MIM 260920) is a rare, apparently monogenic, autosomal recessive disorder characterized by recurrent episodes of fever accompanied with lymphadenopathy, abdominal distress, joint involvement and skin lesions. All patients have high serum IgD values (>100 U/ml) and HIDS 'attacks' are associated with an intense acute phase reaction whose exact pathophysiology remains obscure. Two other hereditary febrile disorders have been described. Familial Mediterranean fever (MIM 249100) is an autosomal recessive disorder affecting mostly populations from the Mediterranean basin and is caused by mutations in the gene MEFV (refs 5,6). Familial Hibernian fever (MIM 142680), also known as autosomal dominant familial recurrent fever, is caused by missense mutations in the gene encoding type I tumour necrosis factor receptor. Here we perform a genome-wide search to map the HIDS gene. Haplotype analysis placed the gene at 12q24 between D12S330 and D12S79. We identified the gene MVK, encoding mevalonate kinase (MK, ATP:mevalonate 5-phosphotransferase; EC 2.7.1.36), as a candidate gene. We characterized 3 missense mutations, a 92-bp loss stemming from a deletion or from exon skipping, and the absence of expression of one allele. Functional analysis demonstrated diminished MK activity in fibroblasts from HIDS patients. Our data establish MVK as the gene responsible for HIDS. PMID- 10369263 TI - The gene mutated in bare patches and striated mice encodes a novel 3beta hydroxysteroid dehydrogenase. AB - X-linked dominant disorders that are exclusively lethal prenatally in hemizygous males have been described in human and mouse. None of the genes responsible has been isolated in either species. The bare patches (Bpa) and striated (Str) mouse mutations were originally identified in female offspring of X-irradiated males. Subsequently, additional independent alleles were described. We have previously mapped these X-linked dominant, male-lethal mutations to an overlapping region of 600 kb that is homologous to human Xq28 (ref. 4) and identified several candidate genes in this interval. Here we report mutations in one of these genes, Nsdhl, encoding an NAD(P)H steroid dehydrogenase-like protein, in two independent Bpa and three independent Str alleles. Quantitative analysis of sterols from tissues of affected Bpa mice support a role for Nsdhl in cholesterol biosynthesis. Our results demonstrate that Bpa and Str are allelic mutations and identify the first mammalian locus associated with an X-linked dominant, male-lethal phenotype. They also expand the spectrum of phenotypes associated with abnormalities of cholesterol metabolism. PMID- 10369264 TI - Mutations in the gene encoding 11-cis retinol dehydrogenase cause delayed dark adaptation and fundus albipunctatus. AB - The metabolic pathways that produce 11-cis retinal are important for vision because this retinoid is the chromophore residing in rhodopsin and the cone opsins. The all-trans retinal that is generated after cone and rod photopigments absorb photons of light is recycled back to 11-cis retinal by the retinal pigment epithelium and Muller cells of the retina. Several of the enzymes involved have recently been purified and molecularly cloned; here we focus on 11-cis retinol dehydrogenase (encoded by the gene RDH5; chromosome 12q13-14; ref. 4), the first cloned enzyme in this pathway. This microsomal enzyme is abundant in the retinal pigment epithelium, where it has been proposed to catalyse the conversion of 11 cis retinol to 11-cis retinal. We evaluated patients with hereditary retinal diseases featuring subretinal spots (retinitis punctata albescens and fundus albipunctatus) and patients with typical dominant or recessive retinitis pigmentosa for mutations in RDH5. Mutations were found only in two unrelated patients, both with fundus albipunctatus; they segregated with disease in the respective families. Recombinant mutant 11-cis retinol dehydrogenases had reduced activity compared with recombinant enzyme with wild-type sequence. Our results suggest that mutant alleles in RDH5 are a cause of fundus albipunctatus, a rare form of stationary night blindness characterized by a delay in the regeneration of cone and rod photopigments. PMID- 10369265 TI - Deafness and imbalance associated with inactivation of the secretory Na-K-2Cl co transporter. AB - Deafness can result from a variety of gene defects. Some genes involved in the physiology of hearing encode membrane transporters that regulate the ionic composition of the fluid bathing the inner ear. The endolymph is an extracellular fluid with an atypical composition that resembles the intracellular milieu, high in K+ and low in Na+. Recent studies have emphasized the prominent role of K+ channels in endolymph secretion and mechanical transduction. Coupled electroneutral transport of Na+, K+ and Cl- is mediated by two isoforms of the Na K-2Cl co-transporter: the absorptive isoform BSC1 (also called NKCC2, encoded by Slc12a1 in mouse) that is exclusively expressed in kidney; and BSC2/NKCC1 (encoded by Slc12a2 in mouse), the secretory isoform which has a wider pattern of expression including epithelia, muscle cells, neurons and red blood cells. These co-transporters share 57% homology at the amino acid level and are pharmacologically inhibited by loop diuretics. There is functional and histochemical evidence for the presence of the secretory isoform of the Na-K-2Cl co-transporter in gerbil, rat and rabbit inner ear. We disrupted mouse Slc12a2 and report here that Slc12a2-/- mice are deaf and exhibit classic shaker/waltzer behaviour, indicative of inner-ear defects. We localized the co-transporter to key secreting epithelia of the mouse inner ear and show that absence of functional co-transporter leads to structural damages in the inner ear consistent with a decrease in endolymph secretion. PMID- 10369266 TI - Mutations in the homeodomain of the human SIX3 gene cause holoprosencephaly. AB - Holoprosencephaly (HPE) is a common, severe malformation of the brain that involves separation of the central nervous system into left and right halves. Mild HPE can consist of signs such as a single central incisor, hypotelorism, microcephaly, or other craniofacial findings that can be present with or without associated brain malformations. The aetiology of HPE is extremely heterogeneous, with the proposed participation of a minimum of 12 HPE-associated genetic loci as well as the causal involvement of specific teratogens acting at the earliest stages of neurulation. The HPE2 locus was recently characterized as a 1-Mb interval on human chromosome 2p21 that contained a gene associated with HPE. A minimal critical region was defined by a set of six overlapping deletions and three clustered translocations in HPE patients. We describe here the isolation and characterization of the human homeobox-containing SIX3 gene from the HPE2 minimal critical region (MCR). We show that at least 2 of the HPE-associated translocation breakpoints in 2p21 are less than 200 kb from the 5' end of SIX3. Mutational analysis has identified four different mutations in the homeodomain of SIX3 that are predicted to interfere with transcriptional activation and are associated with HPE. We propose that SIX3 is the HPE2 gene, essential for the development of the anterior neural plate and eye in humans. PMID- 10369267 TI - A single EFEMP1 mutation associated with both Malattia Leventinese and Doyne honeycomb retinal dystrophy. AB - Malattia Leventinese (ML) and Doyne honeycomb retinal dystrophy (DHRD) refer to two autosomal dominant diseases characterized by yellow-white deposits known as drusen that accumulate beneath the retinal pigment epithelium (RPE). Both loci were mapped to chromosome 2p16-21 (refs 5,6) and this genetic interval has been subsequently narrowed. The importance of these diseases is due in large part to their close phenotypic similarity to age-related macular degeneration (AMD), a disorder with a strong genetic component that accounts for approximately 50% of registered blindness in the Western world. Just as in ML and DHRD, the early hallmark of AMD is the presence of drusen. Here we use a combination of positional and candidate gene methods to identify a single non-conservative mutation (Arg345Trp) in the gene EFEMP1 (for EGF-containing fibrillin-like extracellular matrix protein 1) in all families studied. This change was not present in 477 control individuals or in 494 patients with age-related macular degeneration. Identification of this mutation may aid in the development of an animal model for drusen, as well as in the identification of other genes involved in human macular degeneration. PMID- 10369268 TI - DNA methylation represses transcription in vivo. AB - DNA in somatic tissue is characterized by a bimodal pattern of methylation, which is established in the animal through a series of developmental events. In the mouse blastula, most DNA is unmethylated, but after implantation a wave of de novo methylation modifies most of the genome, excluding the majority of CpG islands, which are mainly associated with housekeeping genes. This genomic methylation pattern is broadly maintained during the life of the organism by maintenance methylation, and generally correlates with gene expression. Experiments both in vitro and in vivo indicate that methylation inhibits transcription. It has not yet been possible, however, to determine the role of DNA methylation on specific sequences during normal development. Cis-acting regulatory elements and trans-acting factors appear to be involved in both stage- and tissue-specific demethylation processes. Sp1-like elements have a key role in protecting the CpG island of Aprt (encoding adenine phosphoribosyltransferase) from de novo methylation, and when these elements are specifically mutated, the Aprt CpG island becomes methylated in transgenic mice. We have now characterized an embryo-specific element from the CpG island sequence upstream of Aprt that can protect itself from de novo methylation in transgenic mice as well as reduce methylation of flanking sequences. We placed this element on a removable cassette adjacent to a human HBB (encoding beta-globin) reporter and generated a transgene whose methylation pattern can be switched in vivo. Analysis of globin transcription in this system showed that methylation in cis inhibits gene expression in a variety of tissues, indicating that DNA modification may serve as a global genomic repressor. PMID- 10369269 TI - Vocal fold proteoglycans and their influence on biomechanics. AB - OBJECTIVE/HYPOTHESIS: To examine the interstitial proteins of the vocal fold and their influence on the biomechanical properties of that tissue. STUDY DESIGN: Anatomic study of the lamina propria of human cadaveric vocal folds combined with some viscosity testing. METHODS: Identification of proteoglycans is performed with histochemical staining. Quantitative analysis is performed using an image analysis system. A rheometer is used for viscosity testing. Three-dimensional rendering program is used for the computer images. RESULTS: Proteoglycans play an important role in tissue biomechanics. Hyaluronic acid is a key molecule that affects viscosity. DISCUSSION: The proteoglycans of the lamina propria have important biological and biomechanical effects. The role of hyaluronic acid in determining tissue viscosity is emphasized. Viscosity, its effect on phonatory threshold pressure and energy expended due to phonation is discussed. CONCLUSION: Proteoglycans, particularly hyaluronic acid, play important roles in determining biomechanical properties of tissue oscillation. Future research will likely make these proteins of important therapeutic interest. PMID- 10369270 TI - Botulinum toxin: adjunctive treatment for posterior glottic synechiae. AB - INTRODUCTION: Synechiae formation of the posterior glottis can result in tracheostomy dependence secondary to airway obstruction. Stenosis is caused by total or partial fixation of the vocal folds in adduction resulting from scar contracture. The treatment poses a management dilemma because of recurrent scar formation, made worse by mobility of the vocal folds. Although various treatment options from conservative endoscopic repair to open procedures have been proposed, the results are not satisfactory and patients often require multiple procedures. METHODS: We present the trial of a conservative approach that includes microscopic CO2 laser resection of the scar with concomitant botulinum toxin injection of the interarytenoid and thyroarytenoid muscles of the more mobile cord. This results in a temporary paresis of the adductor muscles and hence prevents overadduction in the posterior commissure during the postoperative healing period. STUDY DESIGN: We present the surgical technique and results in three patients who underwent the procedure. RESULTS: Treatment in all three patients was successful. CONCLUSIONS: The appropriate use of botulinum toxin may help improve the treatment outcome of posterior synechiae of the larynx without sacrificing any laryngeal components. PMID- 10369271 TI - Zenker's diverticulum: analysis of surgical complications from diverticulectomy and cricopharyngeal myotomy. AB - OBJECTIVE: To identify risk factors for postoperative complications in patients undergoing diverticulectomy and cricopharyngeal (CP) myotomy for Zenker's diverticulum. STUDY DESIGN: Retrospective. MATERIALS AND METHODS: A chart review was conducted of all patients with a Zenker's diverticulum who were treated with diverticulectomy and cricopharyngeal myotomy at three tertiary care centers in central Indiana between 1988 and 1998. RESULTS: Of the 24 patients identified, 9 developed postoperative complications (2 medical and 7 surgical). Statistical analysis of multiple potential risk factors revealed that only diverticulum size greater than 10 cm2 at surgery placed the patient at increased risk for postoperative surgical complications. To our knowledge, this is the first report that has specifically addressed diverticulum size as an independent risk factor for postoperative surgical complications following diverticulectomy and CP myotomy. CONCLUSIONS: Given our findings, we recommend considering diverticulopexy rather than diverticulectomy in a patient with a Zenker's diverticulum greater than 10 cm2 in size if a cervical approach is the selected treatment. PMID- 10369272 TI - Magnetic resonance navigation for head and neck lesions. AB - OBJECTIVE: Review applications of interventional magnetic resonance imaging and describe methods, procedures, and additional instrumentation for the magnetic resonance "operating theater." Describe advantages of magnetic resonance navigation for biopsies of head and neck tumors. STUDY DESIGN: Patients with palpable and nonpalpable head and neck and cranial base tumors were recruited into the study. Patients underwent magnetic resonance-guided biopsy. Retrospective analysis of 21 patients was conducted. METHODS: 0.5 Tesla superconducting open magnetic resonance imaging was used for navigation of the biopsy needle. Patient records and magnetic resonance images were reviewed. The type, size, and location of the lesions were tabulated. Type of anesthesia and monitoring method were analyzed. The histopathologic correlation was conducted in patients who required further surgeries or open surgical biopsies. RESULTS: Twenty-two biopsies were carried out in the magnetic resonance suite. One patient required general anesthesia and the other biopsies were conducted under intravenous sedation. There was only one case of nonconcurrence in a patient with Wegener's granulomatosis of the posterior orbit. Overall, a 92% concurrence rate between image-directed fine-needle aspiration, open biopsy, and surgical therapy was encountered. No complications occurred. CONCLUSIONS: The use of interventional magnetic resonance imaging to assist with fine-needle aspiration core biopsy has made the biopsy procedure safer and more accurate. Potentially morbid and disfiguring surgeries have been avoided in some patients. Deeper lesions have been more easily approached, as the needle for biopsy is under constant magnetic resonance guidance. Improved visualization for critical structures allows safer performance of biopsies. The primary difficulties of open magnetic resonance imaging relate to the need for nonferromagnetic instrumentation and equipment and their high costs. An inverse relationship exists between the imaging quality and the "dead time" required to acquire images. PMID- 10369273 TI - Surgical management of thyroid masses: assessing the need for frozen section evaluation. AB - OBJECTIVE/HYPOTHESIS: To determine the need for intraoperative frozen section to guide the extent of thyroid surgery in the presence of an adequate preoperative fine-needle aspiration (FNA) finding. METHODS: Charts of patients who presented from 1995 to 1998 to the two senior authors were reviewed. A total of 82 patients were found who satisfied the inclusion criteria of having both an adequate FNA and frozen section. The extent of surgery was based on the frozen section finding for all the patients in this study. The authors looked at the number of cases in which the surgical management would be changed if the frozen section was not obtained and the surgical decision was based only on preoperative FNA and intraoperative findings. RESULTS: FNA revealed papillary carcinoma in 18 patients that was confirmed by intraoperative frozen section and final pathology. In the remaining 64 patients, the FNA diagnosis was either benign or suspicious. When routine frozen section was done, 61 of these 64 patients were found to have either benign pathology or pathological diagnosis that was deferred to permanent section. Only three patients were found to have malignancy on frozen section that was missed by FNA. Of these three patients, two had obvious findings of malignancy at the time of surgery. This leaves only one patient with carcinoma that was missed by FNA and intraoperative findings but detected by the intraoperative frozen section. CONCLUSION: Of the 82 patients in this study, only one extra case of malignancy would be missed by elimination of the routine use of intraoperative frozen section. The authors conclude that the routine use of intraoperative frozen section may be unnecessary. The use of an adequate preoperative FNA together with sound clinical judgment at time of surgery can adequately guide the extent of surgical resection. PMID- 10369274 TI - Clinical application of fluorescence in situ hybridization for chromosome 11q13 analysis in head and neck cancer. AB - OBJECTIVE: Squamous cell carcinoma of the head and neck (HNSCC) still has one of the lowest 5-year survival rates. Despite advances in diagnosis, treatment, and research, survival rates have not improved in recent years. This report examines the utility of fluorescence in situ hybridization (FISH) in detecting chromosome 11q13 amplification in HNSCC and in evaluating the correlation between 11q13 amplification and tumor behavior. STUDY DESIGN: This study used FISH to determine the incidence of 11q13 amplification in 20 HNSCCs and 10 normal controls from the same patients. Tumor touch preparations and paraffin-embedded tissues from the same patient samples were used for comparative analysis. Both single and dual color FISH was performed. METHODS: Repetitive chromosome 11 specific alpha satellite DNA probe and chromosome 11q13 specific probe cyclin D1 were used for the FISH analysis. RESULTS: Experiments revealed amplification of chromosome 11q13 in three fresh touch preparations. FISH on paraffin tissues showed amplification in two additional samples. Intensity of amplification, as high as 20 copies per nucleus, was observed in paraffin preparations, whereas a maximum of only six copies was seen in fresh preparations. Amplification was not detected in any of the normal samples. All five cases with 11q13 amplification had metastases and four of these were from poorly differentiated tumors. In the nonamplified cases, 5 of 15 had metastases and 2 of 15 was poorly differentiated. CONCLUSIONS: The present study indicates that FISH is a useful technique for detecting molecular changes such as amplification of chromosome 11q13 in HNSCC. FISH in paraffin preparations allows for accurate measurement of intensity of amplification and makes it possible for the evaluation of a large collection of archival material. The data also suggest that 11q13 amplification is correlated with poorly differentiated tumors and metastasis. Thus FISH has the potential to be a valuable diagnostic/prognostic tool in head and neck cancers. PMID- 10369275 TI - Uses and limitations of FDG positron emission tomography in patients with head and neck cancer. AB - OBJECTIVE: Numerous authors have reported the potential usefulness of positron emission tomography (PET). These studies have had conflicting results, at least partly owing to limited sample sizes. The objective of this study is to define not only the uses, but also the limitations of PET in patients with head and neck cancer. STUDY DESIGN: Nonrandomized, retrospective analysis of PET at an academic institution. METHODS: The authors performed 146 PET scans on 133 patients with head and neck cancer. Eighteen patients (19 PET scans) with thyroid disorders were excluded. A minimum 1 year of follow-up was available in 84 patients, who were separated into groups based on whether the PET was used to detect unknown primary cancers (n = 20), stage neck nodal and distant metastases (n = 8), monitor response to nonsurgical therapy (n = 22), or detect recurrent or residual cancers (n = 34). The results of PET were compared with results from computed tomography (CT) and magnetic resonance imaging (MRI) performed in the same patients. RESULTS: Of the unknown primary cancers, PET correctly identified 7 of 20 primary sites, giving a sensitivity of 35%. When combined with CT or MRI, the sensitivity increased to 40%. When used for detection of metastatic disease, PET demonstrated five of five nodal metastases (100%) and two of four distant metastases (50%). In evaluating the response to nonsurgical therapy, PET had a sensitivity of 50% and a specificity of 83% for detecting tumor at the primary site and a sensitivity of 86% and a specificity of 73% for detecting nodal disease. When used for evaluation of recurrent/residual disease, PET identified seven of seven cases of local recurrences/residual disease and had a specificity of 85%. PET also detected seven of seven cases of nodal disease and had a specificity of 89%. CONCLUSIONS: For staging purposes, PET is limited by its lack of anatomic detail. However, PET compares favorably with CT and MRI in detecting recurrent/residual cancers. PET imaging complements the more traditional imaging modalities (CT or MRI), especially for an unknown primary cancer. PMID- 10369276 TI - Effect of photodynamic therapy on the critical primary ischemic time of fasciocutaneous flaps. AB - BACKGROUND: Photodynamic therapy (PDT) may be used as an adjuvant intraoperative therapy to improve locoregional control. PDT has been shown to delay wound healing. This raises concern about PDTs effect on survival of fasciocutaneous flaps. OBJECTIVE: Evaluate the effect of 1) PDT on the critical ischemic time in a rat fasciocutaneous flap model and 2) photosensitizer activation by the surgical light source. DESIGN: A fasciocutaneous flap, based on the left inferior epigastric vessels, was used. Ischemic times of 2, 4, 6, 8, 10, and 12 hours were induced by clamping the vascular pedicle. Animals were randomly divided into five groups: ischemia only, group I; light treatment to wound bed, group II; Photofrin before surgery with the flap elevated without a fiber optic head light, group III, or with a headlight, group IV; Photofrin prior to surgery with light treatment to the wound bed, group V. Flap survival was assessed on postoperative day 7. RESULTS: The critical primary ischemic time of group V (PDT) was significantly less (P < .05) than groups I, II, III, and IV. There was no statistical difference in the critical primary ischemic time when a fiber optic headlight was used (group III vs. group IV). CONCLUSION: Intraoperative PDT significantly reduces the critical primary ischemic time of the rat fasciocutaneous flap. White light illumination of the operative field does not result in photosensitizer activation and has no effect on the critical primary ischemic time. PMID- 10369277 TI - Open bedside tracheotomy in the intensive care unit. AB - OBJECTIVE: To demonstrate that open bedside tracheotomy is an efficient, safe, and cost-effective procedure. STUDY DESIGN: Retrospective review of more than 200 open bedside tracheotomies performed at UCLA Medical Center, Harbor-UCLA Medical Center, and West Los Angeles VA Medical Center from 1995 to 1998. METHODS: The only personnel required for the procedure were an attending or senior resident and a junior resident or intern, as well as the respiratory therapist to withdraw the endotracheal tube. No anesthetist or scrub nurse was present for any of the procedures. The procedure took an average of 15 to 25 minutes. Patients were followed for 30 days after surgery to determine the incidence of complications. RESULTS: The incidence of major complications related to the procedure, including hemorrhage and myocardial infarction, was less than 1%. The incidence of minor complications, including moderate bleeding at the tracheotomy site, was 4%. Overall mortality within 30 days was 8%, but was not related to the tracheotomy for any patients in this series. The charge for the procedure was $233 for the tracheotomy tube supplies and instruments. This cost compares favorably with an average charge of more than $3000 for the procedure in the operating room and about $1000 for a percutaneous tracheotomy kit. CONCLUSION: Review of our experience demonstrates that open bedside tracheotomies can be performed more efficiently and economically than operating room tracheotomies. The safety of this procedure is comparable to percutaneous tracheotomy but at a decreased cost. PMID- 10369278 TI - Facial nerve rehabilitation after radical parotidectomy. AB - OBJECTIVE: Examine functional outcomes in patients undergoing radical parotidectomy and facial nerve grafting. Identify factors that may affect rehabilitation in these patients. STUDY DESIGN: Retrospective chart review and photographic analyses of 12 patients undergoing radical parotidectomy with interposition nerve grafts for facial nerve reconstruction. METHODS: Data obtained for each patient regarding age, sex, histology of parotid neoplasm, cable graft source, administration of postoperative radiotherapy, and treatment for eye rehabilitation. Functional outcomes were assessed with the House Brackmann grading system at 6 months, 1 year, and 2 years after surgery. RESULTS: All nerve grafts were harvested from cervical plexus sensory nerves with microscopic epineural repair performed for all neurorrhaphies. Overall, 9 of 12 patients achieved a grade III 2 years after surgery. All patients under age 30 obtained a grade III. Of the seven patients receiving postoperative radiation, five achieved a grade III. Older patients often required surgical procedures to facilitate eye closure. CONCLUSIONS: Facial nerve rehabilitation after radical parotidectomy can be successfully achieved with cervical plexus interposition nerve grafts. Postoperative radiotherapy did not appear to affect return of function, and younger patients consistently achieved good functional outcomes after nerve grafting. Older patients frequently require surgical procedures for eye rehabilitation after radical parotidectomy. PMID- 10369279 TI - Success of the modified Epley maneuver in treating benign paroxysmal positional vertigo. AB - OBJECTIVE: Benign paroxysmal positional vertigo (BPPV) is a common condition seen by otolaryngologists. The purpose of this study is to determine the ability of the modified Epley maneuver to treat BPPV. STUDY DESIGN: Retrospective review. METHODS: A retrospective chart review of 107 patients diagnosed with BPPV at our institution between March of 1993 and June of 1995. Each patient was diagnosed with isolated BPPV by history and Hallpike-Dix maneuver. There were no other vestibular symptoms or electronystagmogram abnormalities. Patients diagnosed with BPPV received modified Epley maneuvers, were instructed to remain upright for 48 hours, and wore a soft collar for a week. Patients were followed up with repeat Hallpike-Dix maneuvers at 1 to 2 weeks. If symptoms persisted, the maneuver was repeated for up to a maximum of three times, at which point patients were considered to have failed treatment. RESULTS: The average age of patients was 57.8 years old. Thirty percent were male and the right ear was affected in 54%. The posterior semicircular canal was affected in 105 ears. The average patient received 1.23 Epley maneuvers, with a success rate of 93.4%. No successfully treated patients received mastoid vibration. Seven out of 107 patients failed after three Epley maneuvers. Two failure patients had a history of temporal bone fracture. Two failure patients were treated with posterior semicircular canal block surgery. CONCLUSION: The modified Epley maneuver is an excellent treatment for BPPV. PMID- 10369280 TI - Acoustic neuroma surgery: absent auditory brainstem response does not contraindicate attempted hearing preservation. AB - OBJECTIVE: Absence of auditory brainstem response (ABR) waveforms has been associated with a poor likelihood of hearing preservation following resection of acoustic neuromas. Our experience is reviewed for patients with absent preoperative ABR regarding hearing preservation, hearing improvement, and return of ABR. STUDY DESIGN: Retrospective review of 22 cases of acoustic neuroma resection. Nine patients with absent preoperative ABR were identified. All underwent tumor resection utilizing intraoperative cochlear nerve action potential (CNAP) monitoring. Postoperative hearing results and ABR waveforms were examined. METHODS: Charts were reviewed and tabulated for age, sex, tumor side, tumor size, preoperative and postoperative audiometric and ABR results, intraoperative monitoring results by ABR and CNAP, and surgical complications. RESULTS: Hearing preservation was achieved in seven of nine patients (78%) with absent preoperative ABR, as well as six of seven patients (86%) with tumors less than or equal to 20 mm in greatest dimension. Although intraoperative ABR monitoring was not possible in any of these patients, CNAP monitoring was successful in all. Return of ABR waveforms was observed in four of the six patients (67%) tested from 3 to 22 months postoperatively. Four of the seven patients (57%) enjoyed improvement in hearing class as defined by the guidelines of the American Academy of Otolaryngology-Head and Neck Surgery. CONCLUSIONS: Absent ABR waveforms have not been a negative prognostic sign regarding hearing preservation. CNAP monitoring is possible in these patients and likely helps to minimize iatrogenic cochlear nerve trauma. Patients with no ABR waveforms have hope of hearing preservation and even improvement following acoustic neuroma resection performed utilizing CNAP monitoring and hearing preservation surgical techniques. PMID- 10369281 TI - Double cartilage block ossiculoplasty in chronic ear surgery. AB - OBJECTIVES/HYPOTHESIS: Multiple techniques of ossicular reconstruction have been advocated for hearing rehabilitation in the setting of chronic otitis media No single method can adequately address the clinical spectrum of disease severity. In the situation of a severely diseased ear requiring a canal wall down (CWD) mastoidectomy in the presence of an intact stapes superstructure, the authors have employed a double cartilage block (DCB) ossiculoplasty. The technique and short-term results are reviewed. STUDY DESIGN: Retrospective chart review in a tertiary referral otologic practice. METHODS: Twenty-three patients underwent a CWD tympanomastoidectomy with DCB ossiculoplasty. Ages ranged from 6 to 85 years (mean, 36.1 y). The majority of ears were actively draining at the time of surgery (83%) and most procedures were revisions of prior mastoidectomies (74%). Audiometric data (mean postoperative follow-up, 19.5 mo) were calculated according to 1995 American Academy of Otolaryngology-Head and Neck Surgery guidelines. RESULTS: Audiometric results were available in 20 patients. The mean air-bone gap (ABG) was 23.8 dB after surgery. Closure of the ABG to within 20 dB was achieved in 10 of 20 patients (50%). No cases of DCB extrusion have occurred to date. CONCLUSIONS: The DCB represents an excellent alternative to biocompatible prostheses for ossicular reconstruction in the setting of severe chronic ear disease. As with all methods of ossiculoplasty, long-term follow-up will be necessary to determine if this technique remains stable in the hostile environment in which it has been employed. PMID- 10369282 TI - Reduced postoperative infections with an immune-enhancing nutritional supplement. AB - OBJECTIVES/HYPOTHESIS: Malnutrition is a significant risk factor for postoperative infections in patients undergoing oncologic surgery. This study was undertaken to determine if perioperative nutritional supplementation with an immune-enhancing formula is superior to standard formula in the prevention of postoperative infectious complications. STUDY DESIGN: This was a prospective, randomized, double-blind trial comparing perioperative nutritional supplementation with Impact and standard nutritional formulas. METHODS: Following stratification, 136 patients undergoing oncologic head and neck surgery were randomly assigned to one of four treatment groups: preoperative/postoperative Impact, postoperative Impact, preoperative/postoperative standard formula, and postoperative standard formula. Outcome measures included laboratory evaluations of nutritional status, infectious and wound healing complications, and duration of hospitalization. Statistical analysis was performed using chi2 or two-tailed Fisher Exact Tests, when appropriate. RESULTS: Intent-to-treat (P = .02) and actual therapy (P = .04) analyses revealed a significant decrease in the incidence of postoperative infectious complications (all sites) in patients who received Impact. There was no significant difference in wound healing problems or duration of hospitalization. Postoperative measures of nutrition status demonstrated a higher serum albumin (P = .05) in patients who received Impact compared with standard formula. CONCLUSIONS: Compared with standard formula, perioperative nutritional supplementation with Impact significantly reduced the incidence of infectious complications. The length of hospitalization was significantly prolonged in patients with postoperative infections, suggesting potential cost savings with the use of immune-enhancing formulas such as Impact. PMID- 10369283 TI - Intraoperative electromyography for predicting facial function in vestibular schwannoma surgery. AB - OBJECTIVE: To assess the validity of intraoperative minimal stimulation threshold (MST) for predicting long-term facial function after vestibular schwannoma surgery. STUDY DESIGN: Prospective blinded study. METHODS: MST after tumor dissection and postoperative clinical facial function, assessed using the House Brackmann grading system (HB), were used to predict long-term clinical facial function, recorded at least 6 months after surgery. RESULTS: Two hundred and nine consecutive patients fulfilled selection criteria and 184 had successful intraoperative electrophysiologic monitoring and were eligible for further study. MST of 0.05 mA had moderate accuracy for predicting good long-term facial function, with 94% sensitivity, 91% positive predictive value (PPV), 60% specificity, and 70% negative predictive value (NPV). A more relevant group of 77 patients with poor postoperative facial function (HB III-VI) were assessed for predicting good long-term function. Applying this criteria, test accuracy fell, with 83% sensitivity, 64% PPV, 60% specificity, and 75% NPV. Postoperative clinical facial function had a greater accuracy for predicting good long-term function, with 83% sensitivity, 79% PPV, 75% specificity, and 79% NPV. A model of predicted probabilities of good outcome (HB I and II) was derived from a logistic regression with two additive predictors (postoperative HB and MST). This demonstrated that for patients with postoperative HB grade V, MST aided prediction. CONCLUSIONS: Intraoperative stimulation thresholds, when assessed against a relevant group of patients with poor postoperative facial function, had poor predictive accuracy. The severity of immediate postoperative clinical facial function was the most accurate predictor of long-term outcome. MST aided long term prediction in a small but relevant group of patients with postoperative HB grade V facial function. PMID- 10369284 TI - Human temporal bone study on the postnatal ossification process of auditory ossicles. AB - OBJECTIVE: In infancy the head of the malleus and body of the incus normally contain bone marrow, which is gradually replaced by bone and converted into vascular channels with age. This study was carried out to clarify the age at which ossification of the ossicles is complete and to examine factors influencing the ossification process. STUDY DESIGN: Human temporal bone sections from 32 infants and children with or without congenital anomalies, aged 1 day to 9 years, who were born at term were studied. METHODS: The percentage bone marrow area occupying the head of the malleus and body of the incus was calculated in three horizontal temporal bone sections for each case, using computer-aided digital processing of images. RESULTS: Bone marrow was observed in both the malleus and incus in children until 25 months of age, while after the age of 25 months no bone marrow tissue was present in either of the ossicles. It appeared that the bone marrow space disappeared somewhat earlier in the malleus than in the incus. The bone marrow space around the otic capsule disappeared much earlier than that within the ossicles. The age at completion of ossification was correlated with neither the presence of congenital anomalies nor the presence of residual mesenchyme in the middle ear. CONCLUSIONS: Ossification of the ossicles seems to occur steadily throughout fetal life and after birth during development of the middle ear. Although the clinical significance of postnatal residual bone marrow within the ossicles is not known, it possibly plays a role as a blood-forming organ in early infancy. PMID- 10369285 TI - Chronology of labyrinthitis ossificans induced by Streptococcus pneumoniae meningitis. AB - OBJECTIVE: Labyrinthitis ossificans consists of novel osteogenesis that fills the normally patent cochlear and vestibular lumen as an end-stage sequelae to various pathologies. This study was designed to establish the sequence of events and chronology of the osteoneogenesis and calcification. STUDY DESIGN: A prospective randomized double-blind study. METHODS: By using serial application of different colored fluorochromes, which deposit in newly forming bone, the timing of bone deposition and bone remodeling can be established. Labyrinthitis ossificans was induced in six groups (n = 5) of gerbils by an intrathecal injection of live Streptococcus pneumoniae. Group 1 received no fluorochrome labels, group 2 received one label, group 3 received three labels, and groups 4, 5, and 6 received four labels. The temporal bones were harvested after 2 weeks (group 1), 1 month (group 2), 3 months (group 3), 4 months (group 4), 6 months (group 5), and 12 months (group 6). RESULTS: Sixteen of the 25 animals that received labels developed ossification, demonstrated with fluorescent microscopy. In the animals that developed labyrinthitis ossificans, newly formed disorganized bone began calcifying as early as 3 weeks (label 1) after S. pneumoniae injection. Osteoneogenesis continued as evidenced by the presence of the other labels when first applied at 6 weeks (label 2), and 10 weeks (label 3). Ossification, calcification, and remodeling proceeded through a 12-month course, wherein a reduction of labels was present at 6 months and total disappearance by 12 months. CONCLUSIONS: The use of fluorescent stains in this animal model provides a means to establish a timeline of the ossification seen in labyrinthitis ossificans. PMID- 10369286 TI - Influence of lateral osteotomies in the dimensions of the nasal cavity. AB - OBJECTIVES: To elucidate the importance of placement of lateral osteotomy in rhinoplasty at a level above or below the insertion of the inferior turbinate at the pyriform aperture. STUDY DESIGN: Controlled lateral osteotomies were performed in 16 cadaver noses. Eight of the lateral ostetomies were placed below (low) and eight were placed above (high) the insertion of the inferior turbinate. In all 16 noses medial osteotomies were performed. Dimensions of the nasal cavity were measured by acoustic rhinometry before and after the osteotomies. METHODS: The total minimum cross-sectional area (TMCA) and the cross-sectional area at the pyriform aperture (TCA-3.3) were calculated and the preoperative and postoperative values were analyzed statistically. RESULTS: There was no significant difference in the reduction of cross-sectional area in the group that underwent high lateral osteotomy compared with the group the underwent low lateral osteotomy. In both groups the TMCA was reduced, with 12% of the value (P = .001) before osteotomy, and the TCA-3.3 was reduced, with 15% of the value before osteotomy (P = .000). CONCLUSIONS: After lateral and medial ostetomies a significant decrease in the anterior dimensions of the nose is observed. The decrease following osteotomy does not seem to be due to the placement of the lateral osteotomy at the pyriform aperture but rather to the detachment of the bony vault from the surrounding structures. PMID- 10369287 TI - Rhinosinusitis and atopy in patients infected with HIV. AB - HYPOTHESIS: Rhinosinusitis is common during HIV infection; its prevalence is uncertain and could probably be related to clinical features, immunoallergological status, and diagnostic criteria METHODS: Seventy-four patients hospitalized with HIV infection were prospectively evaluated for the presence of rhinosinusitis based on clinical findings, nasal endoscopy, or paranasal sinus computed tomography (CT). Immune status, nasal smear, features of atopy (based on the prick test), and its contribution to sinusal inflammatory pathology were also evaluated. RESULTS: Most patients were severely immunosuppressed: CD4+ 155+/-201 cells/mL and 12+/-11% (mean +/- SD). Thirty-five percent of the patients presented at least two criteria of rhinosinusitis (clinical findings, nasal endoscopy, and CT: 35%; clinical findings and CT: 50%; nasal endoscopy and CT: 15%). CT scan showed multiple sinus involvement, opacification over 25% of the total volume of the maxillary sinus in 50% of patients, and opacification of the sphenoidal sinus in 40% of cases. Atopy was present in 18% of patients, a figure which reflects the expected prevalence in our geographic area. Two independent predictors were associated with a higher probability of rhinosinusitis: bilateral absence of maxillary infundibular patency (odds ratio, 7.5; 95% CI = 2.03-27.9) and low total count (odds ratio, 0.99; 95% CI = 0.99-1.00) or percentage of CD4+ (odds ratio, 0.93; 95% CI = 0.88 1.00). CONCLUSIONS: There is a high prevalence of rhinosinusitis in HIV-infected individuals. This finding is related to a decreased cellular immunity, but it does not appear to be related to IgE-related immediate hypersensitivity. Nasal endoscopy should be the first-step diagnostic test. However, when clinical suspicion exists and endoscopy fails to explain symptoms, CT scan is a valuable adjunct to establish this diagnosis. PMID- 10369288 TI - Postoperative care in functional endoscopic sinus surgery? AB - OBJECTIVE: To assess the value of nonintervention after FESS. STUDY DESIGN: Prospective study. METHODS: Fifty-five patients with diagnosed chronic rhinosinusitis who failed adequate medical therapy were subjected to FESS. No postoperative care, apart from nasal douching with hypertonic saline after the tenth postoperative day, was done. No antibiotics or steroids were administered routinely. Because 10 patients were not available for follow-up, only 45 patients were included in the study. RESULTS: Success rate judged by at least 50% subjective improvement of symptoms was 95.5%. However, all patients derived some benefit. The occurrence of postoperative synechiae is discussed. CONCLUSIONS: The usefulness of postoperative care of the FESS cavity needs reappraisal. PMID- 10369289 TI - Laser management of oral leukoplakias: a follow-up study of 70 patients. AB - OBJECTIVES/HYPOTHESIS: To assess the efficacy of laser therapy for the management of premalignant oral lesions. STUDY DESIGN: The study group consisted of seventy consecutive laser-treated patients with oral leukoplakia. The microscopic diagnosis included idiopathic focal keratosis, dysplasias of all grades, and verrucous hyperplasia (proliferative verrucous leukoplakia). Thirty-nine patients had some degree of microscopic dysplasia and six demonstrated high-risk proliferative verrucous leukoplakia. The clinical appearances of the lesions were white (homogeneous leukoplakia) in 48, red and white (erythroleukoplakia) in 8, and verrucous in 14. There were 38 men and 32 women in this group. The average age was 63 years (range, 31-90 y). METHODS: Lasers employed were the CO2 and Nd:YAG lasers, and standard laser safety protocols were used. RESULTS: There was no postoperative infection, hemorrhage, or paresthesia Two patients developed pyogenic granulomas in their surgical sites. Fifty-five of 70 patients were followed for more than 6 months; follow-up averaged 32 months (range 6-178 mo). Twenty-nine patients had complete control of their lesions; 19 patients had small recurrences removed with subsequent laser surgeries, leading to control; 2 patients had complete recurrences; and 5 patients developed squamous cell carcinoma at the lesion site. Verrucous lesions had an especially high rate of recurrence (83%), with 9 of 12 ultimately controlled with subsequent surgeries. CONCLUSIONS: Laser surgery of oral leukoplakia is an effective tool in a complete management strategy that includes careful clinical follow-up, patient education to eliminate risk factors and report suspicious lesions, and biopsy of suspicious lesions when appropriate. However, recurrence and progression to cancer remain a risk. PMID- 10369290 TI - Quantitative evaluation of the upper airway during nasopharyngoscopy with the Muller maneuver. AB - OBJECTIVE: To quantitatively examine changes in the upper airway caliber of normal subjects at graded negative inspiratory pressures generated during nasopharyngoscopy with a Muller maneuver. STUDY DESIGN: Eighteen normal subjects prospectively underwent nasopharyngoscopy with Muller maneuvers. Subjects performed graded and maximal effort Muller maneuvers while sitting upright, and maximal-effort Muller maneuvers in the supine position. Two regions of the upper airway--the retropalatal and retroglossal--were examined. METHODS: Images from the endoscopic examination were objectively analyzed by adjusting manually traced airway contours using full-width, half-maximum edge detection algorithm software. The adjusted tracings' area and dimensions through the airway centroid were measured. RESULTS: Muller maneuvers performed at -40 cm H2O resulted in a 64%+/ 17% (P = .0001) reduction in upper airway area that consisted of a 51%+/-20% (P = .0001) reduction in the lateral dimension and a 21%+/-24% (P = .0026) reduction in antero-posterior dimension. Muller maneuvers in the retroglossal region did not significantly reduce airway area (P = .575), but demonstrated an altered airway conformation that consisted of lateral narrowing and an increase in antero posterior dimension. Changes in body position did not result in significant differences in either airway caliber or airway dimension. CONCLUSIONS: Airway caliber during forced inspiration is mediated primarily through changes in the lateral pharyngeal walls. This study has also shown that antero-posterior and lateral airway structures are largely independent in their response to Muller maneuvers. Similarly, the retropalatal and retroglossal regions of the upper airway respond differently to forced negative intraluminal pressure. PMID- 10369291 TI - Osteosarcoma of the head and neck: a review of the Johns Hopkins experience. AB - OBJECTIVE: To determine factors including treatment modalities which influence survival in patients with osteosarcoma of the head and neck. STUDY DESIGN: Retrospective clinicopathologic study of 27 patients with osteosarcoma of the head and neck. METHODS: The clinical charts and pathology slides were reviewed on 27 patients who had osteosarcoma of the head and neck between 1946 and 1998. The following variables were examined for their effect on survival: age of diagnosis, site of tumor, presentation, race, sex, prior radiation exposure, retinoblastoma history, margin status, and method of treatment. RESULTS: The average age at the time of diagnosis of the patients was 37.6 years (range, 7-82 y). The sex distribution was similar with 14 male and 13 female patients. Eight of 27 patients had osteosarcoma of the mandible, 9 of 27 had osteosarcoma of the maxilla and paranasal sinuses, and in 10 of 27 patients osteosarcoma occurred elsewhere, including the temporal bones, occipital bones, and orbit. The overall 2-year survival was 66% with a 5-year survival rate of 55%. CONCLUSIONS: Positive surgical margins and a high tumor grade were found to have a statistically deleterious effect on overall survival. There was no detectable effect on survival of age, race, sex, prior radiation exposure, tumor site, and tumor cell type. It was not possible to differentiate between the different adjuvant treatment modalities because of the small numbers in the study. PMID- 10369292 TI - Detection of recurrent head and neck squamous cell carcinomas after radiation therapy with 2-18F-fluoro-2-deoxy-D-glucose positron emission tomography. AB - OBJECTIVES/HYPOTHESIS: Fluorodeoxyglucose positron emission tomography (FDG-PET) has been proposed as a sensitive method to diagnose and stage various malignancies. We assessed the efficacy of FDG-PET imaging in distinguishing tumor persistence/recurrence from posttreatment changes following radiation therapy for squamous carcinomas of the head and neck STUDY DESIGN: Retrospective analysis of FDG-PET results compared with biopsy results or outcome, or both. METHODS: Twenty eight patients who had undergone radiation therapy with or without surgery for treatment of squamous cell carcinoma were studied with FDG-PET imaging. There was clinical suspicion for recurrence in each patient, but no obvious mass or lesion to biopsy was found on physical examination or anatomic imaging. The results of FDG-PET imaging were compared with those of biopsy or clinical follow-up of at least 6 months, or both. RESULTS: FDG-PET imaging was positive in 13 patients, and the presence of active disease was confirmed in 12. Two thirds of the 12 received further cancer treatment. There were 15 negative FDG-PET images. Thirteen of these were confirmed true-negative images, but two studies were false negative images. The sensitivity and specificity of FDG-PET were 86% and 93%, respectively, with positive and negative predictive values of 92% and 87%, respectively. The overall accuracy was 89%. CONCLUSION: FDG-PET imaging is a useful modality to distinguish tumor persistence/recurrence from radiation induced tissue changes in the neck following treatment for head and neck cancer. FDG-PET can identify patients who may benefit from further treatment, and may lead to improved outcome for individual patients. PMID- 10369294 TI - An anatomical study of anastomoses between the laryngeal nerves. AB - OBJECTIVE: To systematize the anatomy of the connecting branches between laryngeal nerves. METHODS: Microdissection of 90 larynges obtained from necropsies (57 men and 33 women; age range, 41-95 y). RESULTS: Anastomoses between the internal and recurrent nerves appeared in four different patterns: 1) Galen's anastomosis, as the connection between the dorsal branches of both nerves (100%); 2) arytenoid plexus, as the connection between the arytenoid branches of both nerves, in relation with the arytenoid muscle, and divided in a deep part (100%) and a superficial part (86%); 3) cricoid anastomosis, previously only described in cows, located in the front of the cricoid lamina (6/10 cases); and 4) thyroarytenoid anastomosis, as the connection of a descending branch of the internal laryngeal nerve and an ascending branch of the recurrent nerve (14%). Anastomosis between the internal laryngeal and the external laryngeal nerves appeared as a connecting branch throughout the foramen thyroideum (21%). Anastomosis between the external laryngeal and recurrent nerves appeared as a connecting branch throughout the cricothyroid muscle (68%). CONCLUSION: At least two anastomoses (Galen's anastomosis and arytenoid plexus) appeared in 21% of hemilarynges, and 79% of cases had three or more anastomoses between the laryngeal nerves. The different prevalence of this complex anastomotic pattern suggests functional differences in the sensory and motor innervation of individual subjects. PMID- 10369293 TI - Establishment and characterization of human laryngeal squamous cell carcinoma cell lines. AB - OBJECTIVES: Six human laryngeal squamous cell carcinoma cell lines (SNU-46, -585, -899, -1066, -1076, -1214) established from Korean patients are reported. STUDY DESIGN: In vitro culture of six squamous cell carcinoma cell lines derived from primary tumors of the larynx. Description of the cell line phenotypes and determination of molecular characteristics. METHODS: Six laryngeal squamous cell carcinoma cell lines were cultured. The cell phenotypes, including the histopathology of the primary tumors and in vitro growth characteristics, were determined. Molecular characterization was also performed, including DNA fingerprinting analysis and abnormalities of p15, p16, p53, and TGF-betaRII genes by polymerase chain reaction-based single strand conformation polymorphism and sequencing analysis. RESULTS: All cell lines grew as adherent cells; five lines grew as monolayers and one other line grew as stratifying colonies. All lines showed 1) high viability (75%-92%) with various doubling times (36-96 h); 2) absence of Mycoplasma and other bacteria; and 3) genetic heterogeneity by DNA profile analysis. p53 Mutations were found in three lines and p16 mutations were observed in five cell lines. TGF-betaRII mutations were found in two lines: one line had frameshift mutation and another line had a missense mutation at the kinase domain. CONCLUSIONS: These newly established and characterized laryngeal squamous cell carcinoma cell lines will be useful for investigating the biologic characteristics of laryngeal cancer. PMID- 10369295 TI - Pulmonary function after total laryngectomy. AB - OBJECTIVE: To present data on pulmonary function in 59 laryngectomees using a specially designed silicone adapter for connection of the stoma to the bodyplethysmograph. STUDY DESIGN: Prospective assessment of pulmonary function in 59 patients, and comparison of lung function before and after bronchodilator testing in selected cases. METHODS: The usefulness of the adapter was examined. Data of patients with airway obstruction were tabulated according to large airway obstruction (LAO), peripheral airway obstruction (PAO), and small airway disease (SAD) types. RESULTS: Findings show that pulmonary airway obstruction was present in 81% of patients and normal pulmonary function was present in only 11 of the 59 participants (19%). LAO was found in 25%, PAO in 17%, and SAD in 39% of cases. Emphysema was diagnosed in 14% of cases within the above-mentioned LAO and PAO groups. Improvement of pulmonary function was achieved in 12 of 16 laryngectomees with airway obstruction, when a bronchodilator aerosol was administered. In 60% of cases with LAO and PAO, the laryngectomees did not know of any marked obstruction of their airways, and only 10% of those knowing about their obstruction received appropriate medical treatment. CONCLUSIONS: These results suggest that 42% of the laryngectomees tested may have benefited from further medical treatment. After laryngectomy, pulmonary function assessment was performed elsewhere in 1 of 59 cases. In light of the high prevalence of airway obstruction in laryngectomees, more frequent postoperative assessments of pulmonary function should be offered to prevent or to reduce impairment of respiratory function in this postlaryngectomy vulnerable pulmonary status. Further studies are also needed to determine the effects of therapeutic intervention, e.g., assessment of therapy outcome and influence on quality of life. PMID- 10369296 TI - Idiopathic bilateral vocal fold weakness. AB - OBJECTIVE: To describe an unrecognized clinical entity, idiopathic bilateral vocal fold weakness, and propose recommendations regarding the diagnosis and management of these cases. STUDY DESIGN: Retrospective, nonrandomized case study. METHODS: All cases of bilateral vocal fold weakness evaluated at the University of Washington Voice Disorders Clinic between 1991 to 1998 were reviewed. RESULTS: Four patients with bilateral laryngeal weakness were determined to have idiopathic bilateral vocal fold paresis following exhaustive workups, including videostroboscopy, bilateral laryngeal electromyography (EMG), neurological consultation, and other pertinent studies. CONCLUSIONS: Performing bilateral laryngeal EMG is an essential aspect of the workup of any laryngeal weakness case, particularly if the etiology is unknown on presentation. Idiopathic bilateral vocal fold weakness is an underrecognized but real clinical diagnosis that will become more familiar with the increasing utilization of laryngeal EMG in clinical situations. PMID- 10369297 TI - Repair of the persistent cerebrospinal fluid leak with the radial forearm free fascial flap. PMID- 10369298 TI - Endoscopic inferior turbinate reduction: a new technique. PMID- 10369299 TI - Endoscope-assisted vestibular neurectomy. PMID- 10369300 TI - RET/PTC fusion gene products in patients suffering from thyroid carcinomas. PMID- 10369301 TI - Cerebrospinal fluid opens a window on Alzheimer disease. PMID- 10369302 TI - Domestic violence in neurologic practice. PMID- 10369303 TI - Mechanisms of high-dose intravenous immunoglobulins in demyelinating diseases. AB - Administration of high-dose intravenous immunoglobulins has become one of the most successful new treatment regimens for demyelinating diseases. In a decade of molecular medicine, it came as a surprise that a natural blood product would prove effective in several disorders, including Guillain-Barre syndrome, chronic inflammatory demyelinating polyneuropathy, multifocal motor neuropathy, and, probably, multiple sclerosis. Many experimental studies, both in vivo and in vitro, have shown that intravenous immunoglobulins can interfere with the immune system at several levels. In addition, intravenous immunoglobulins may promote remyelination in demyelinating disease associated with viral infections. At present, no single mode of action has been identified as the crucial mechanism, which leads us to suggest that multiple effects may act in concert. PMID- 10369304 TI - Genetic linkage analysis. AB - Genetic linkage analysis is a powerful tool to detect the chromosomal location of disease genes. It is based on the observation that genes that reside physically close on a chromosome remain linked during meiosis. For most neurologic diseases for which the underlying biochemical defect was not known, the identification of the chromosomal location of the disease gene was the first step in its eventual isolation. By now, genes that have been isolated in this way include examples from all types of neurologic diseases, from neurodegenerative diseases such as Alzheimer, Parkinson, or ataxias, to diseases of ion channels leading to periodic paralysis or hemiplegic migraine, to tumor syndromes such as neurofibromatosis types 1 and 2. PMID- 10369305 TI - Cerebrospinal fluid beta-amyloid(1-42) in Alzheimer disease: differences between early- and late-onset Alzheimer disease and stability during the course of disease. AB - OBJECTIVES: To study the diagnostic potential of the 42 amino acid form of beta amyloid (beta-amyloid(1-42)) in cerebrospinal fluid (CSF) as a biochemical marker for Alzheimer disease (AD), the intra-individual biological variation of CSF-beta amyloid(1-42) level in patients with AD, and the possible effects of differential binding between beta-amyloid and apolipoprotein E isoforms on CSF-beta-amyloid(1 42) levels. DESIGN: A 20-month prospective follow-up study. SETTING: Community population-based sample of consecutive patients with AD referred to the Pitea River Valley Hospital, Pitea, Sweden. PATIENTS: Fifty-three patients with AD (mean +/- SD age, 71.4 +/- 7.4 years) diagnosed according to the National Institute of Neurological and Communicative Disorders and Stroke and Alzheimer's Disease and Related Disorders Association criteria and 21 healthy, age-matched (mean +/- SD age, 68.8 +/- 8.0 years) control subjects. MAIN OUTCOME MEASURES: Cerebrospinal fluid beta-amyloid(1-42) level--analyzed using enzyme-linked immunosorbent assay--and severity of dementia--analyzed using the Mini-Mental State Examination. RESULTS: Mean +/- SD levels of CSF-beta-amyloid(1-42) were decreased (P<.001) in patients with AD (709 +/- 304 pg/mL) compared with controls (1678 +/- 436 pg/mL). Most patients with AD (49 [92%] of 53 patients) had reduced levels (<1130 pg/mL). A highly significant correlation (r = 0.90; P<.001) between baseline and 1-year follow-up CSF-beta-amyloid(1-42) levels was found. There were no significant correlations between CSF-beta-amyloid(1-42) level and duration (r = -0.16) or severity (r = -0.02) of dementia. Low levels were also found in patients with mild dementia (Mini-Mental State Examination score, >25). CONCLUSIONS: The sensitivity of CSF-beta-amyloid(1-42) level as a diagnostic marker for AD is high. The intra-individual biological variation in CSF-beta amyloid(1-42) level is low. Low CSF-beta-amyloid(1-42) levels are also found in the earlier stages of dementia in patients with AD. These findings suggest that CSF-beta-amyloid(1-42) analyses may be of value in the clinical diagnosis of AD, especially in the early course of the disease, when drug therapy may have the greatest potential of being effective but clinical diagnosis is particularly difficult. PMID- 10369306 TI - Domestic violence against patients with chronic neurologic disorders. AB - BACKGROUND: Violent behavior caused by some neurologic disorders has been widely studied. However, the inverse, violence suffered by patients with neurologic disorders, has not been reported. Brain disorders frequently produce a high frequency of social, psychological, or physical disabilities that could leave patients vulnerable to domestic violence. OBJECTIVES: To determine the prevalence of domestic violence among female patients with chronic neurologic disorders and to identify possible diagnoses associated with the battering syndrome. DESIGN: Cross-sectional, self-administered, anonymous survey. SETTING: Tertiary care center for neurologic disorders in Mexico. PATIENTS: One thousand consecutive adult female patients with neurologic disorders, separated by medical diagnosis of functional or structural disorders. MAIN OUTCOME MEASURES: A modified version of the Abuse Assessment Screen was administered. Statistical analysis was performed using Poisson regression to estimate the prevalence ratio by univariate and multivariate analysis. RESULTS: Overall, 31.2% of women with chronic neurologic disorders were survivors of domestic violence. When separated according to the nature of the disease, 35.3% of patients with functional disorders and 28.1% of patients with brain structural disorders were victims of domestic violence (P = .02). Risk increased in relation to duration of marriage, number of children, and work outside the home. CONCLUSIONS: One third of female patients with chronic neurologic disorders in Mexico suffer domestic violence. A higher frequency of domestic violence was endured by patients with diagnosis of functional disorders as essential epilepsy, headache, migraine, trigeminal pain, depression, or vertigo. The possibility of domestic violence should be routinely explored in patients with chronic neurologic disorders of functional origin. PMID- 10369308 TI - A novel mutation in the gene for the adult skeletal muscle sodium channel alpha subunit (SCN4A) that causes paramyotonia congenita of von Eulenburg. AB - BACKGROUND: Paramyotonia congenita (PMC) of von Eulenburg is an autosomal dominant muscular disease characterized by exercise- and cold-induced myotonia and weakness. To date, 18 missense mutations in the adult skeletal muscle sodium channel alpha-subunit (SCN4A) gene have been identified to cause a spectrum of muscular diseases, including PMC of von Eulenburg, PMC without cold paralysis, potassium-aggravating myotonia, and hyperkalemic periodic paralysis. However, no obvious correlations can be made between the location or nature of amino acid substitutions in SCN4A and its clinical phenotypes. OBJECTIVE: To describe clinical and genetic features of a family with PMC of von Eulenburg. RESULTS: A Japanese family with cold-induced myotonia and weakness was diagnosed as having PMC of von Eulenburg. This phenotype was identified to be caused by a novel mutation that substituted a glutamic acid residue for a highly conserved glycine residue in the fourth transmembrane segment (S4) of domain IV. This predicted a decrease in positive charge specific for the S4. CONCLUSION: In addition to the G1456E identified in this study, 4 mutations that cause a decrease in positive charge in the S4/D4 are associated with the phenotype of PMC of von Eulenburg. This provides an important genotype-phenotype correlation in sodium channelopathies. PMID- 10369307 TI - Urinary myelin basic protein-like material in patients with multiple sclerosis during interferon beta-1b treatment. AB - OBJECTIVES: To determine levels of urinary myelin basic protein-like material (MBPLM) in patients with multiple sclerosis (MS) openly treated with interferon beta-1b and to correlate these with clinical changes. BACKGROUND: Levels of urinary MBPLM correlate with the presence of the progressive phase of MS and with the disease burden detected on T2-weighted, cranial magnetic resonance imaging. Measurement of urinary MBPLM level may be a feasible test for monitoring or predicting response to therapeutic measures. DESIGN AND METHODS: In a prospective study at one site, 166 patients with MS (131 with relapsing-remitting [RR] and 35 with secondary progressive [SP] disease) were treated for a minimum of 1 year and up to 3 years with interferon beta-1b and underwent assessment for neurologic disability (Expanded Disability Status Scale and Scripps Neurological Rating Scale) and change in disease subtype. Urine samples were obtained at 1219 of 1378 clinic visits, and urinary MBPLM level was determined and related to creatinine level to adjust for renal function. RESULTS: Statistical analysis using the general linear models procedure confirmed previous findings that the level of urinary MBPLM related to urinary creatinine level (MBPLM/creatinine) was higher (P<.001) in patients with SP than RR MS. Of the 131 patients with RR MS, SP disease developed in 13 during the observation period. Compared with those in the RR group, the RR to SP group had a higher level (P<.001) of urinary MBPLM and did not differ from the SP group. CONCLUSIONS: The level of urinary MBPLM is higher in SP MS than RR MS but not in RR MS that converts to SP MS. Level of urinary MBPLM may permit the examination of treatment tested to prevent RR disease from becoming progressive. PMID- 10369309 TI - Microalbuminuria in ischemic stroke. AB - OBJECTIVES: To determine (1) the incidence of microalbuminuria in patients with recent ischemic stroke, (2) its relationship to risk factors for stroke, (3) its prevalence in the major subtypes of ischemic stroke, and (4) its potential for identifying patients at increased risk for recurrent stroke, myocardial infarction, or vascular death. DESIGN: Prospective case-control study. SETTING: Outpatient clinics at the medical centers affiliated with the Department of Veterans Affairs and Oregon Health Sciences University in Portland, Ore. PATIENTS: A total of 186 older men and women (median age, 65 years) who were enrolled in a prospective study of risk factors for recurrent stroke, including 97 patients with recent (6-8 weeks) ischemic stroke, 51 with similar clinical risk factors for stroke, including 24 with a history of remote stroke or transient ischemic attack, and 38 community-dwelling volunteers. RESULTS: Microalbuminuria was 3 times more prevalent in patients with recent stroke (29%) than in those with clinical risk factors for stroke (10%), and was undetectable in healthy elderly controls (P<.001). The presence of microalbuminuria in recent stroke as well as in the combined recent and remote stroke or transient ischemic attack group (n = 121) was predicted by diabetes (odds ratio [OR], 8.4; 95% confidence interval [CI], 2.6-27.0; P<.001; serum albumin levels (OR, 0.12; 95% CI, 0.03-0.50; P<.005); age (OR, 1.1; 95% CI, 1.0-1.2; P<.01), and ischemic heart disease (OR, 3.0; 95% CI, 1.0-9.1; P<.05). Among patients with recent stroke the prevalence of microalbuminuria did not differ among major ischemic stroke subtypes, ie, atheroembolic, 23%; cardioembolic, 30%; and lacunar, 33%. During a mean +/- SD of 1.5 +/- 0.9 years of follow-up, 20% of patients with recent stroke, 14% with risk factors for stroke, and 0% of healthy elderly volunteers had vascular end points (P<.004), with events being as frequent in patients with microalbuminuria (32%) as in patients with macroalbuminuria (33%). After controlling for major clinical risk factors, microalbuminuria remained an independently significant predictor of future stroke in the combined recent stroke and remote stroke or transient ischemic attack group (Cox proportional hazard ratio, 4.9; 95% CI, 1.4-17.6; P<.01). CONCLUSIONS: Microalbuminuria is a common finding in patients with cerebrovascular disease and is associated with increased risk for stroke even after correction for the presence of confounding clinical risk factors. These data suggest that microalbuminuria merits further examination as a potentially inexpensive and easily measured marker of increased risk for stroke. PMID- 10369310 TI - Salvage chemotherapy with tamoxifen for recurrent anaplastic astrocytomas. AB - BACKGROUND: A prospective phase 2 study of daily oral tamoxifen citrate in young adults with recurrent anaplastic astrocytomas. METHODS: Twenty-four patients (15 men; 9 women) aged 19 to 45 years (median age, 31.5 years) with recurrent anaplastic astrocytomas were treated. All patients had been treated previously with surgery and involved-field radiotherapy (median dose, 60 Gy; range, 59-61 Gy). In addition, 22 patients were treated adjuvantly with nitrosourea-based chemotherapy (combined procarbazine hydrochloride, lomustine, and vincristine sulfate in 16; carmustine in 6). All patients were treated with salvage chemotherapy at first recurrence, with 1 to 4 chemotherapy regimens (median, 1 regimen). Tamoxifen citrate was administered orally at a fixed dosage of 80 mg/m2 as a single or a twice-daily dosage. Neurologic and neuroradiographic evaluation were performed every 12 weeks, operationally defined as a single cycle of tamoxifen. RESULTS: All patients were able to undergo evaluation. A median of 4 cycles of tamoxifen (range, 1-8 cycles) were administered. No tamoxifen-related toxic effects were seen, nor were there any treatment-related deaths. Four patients (17%) demonstrated a neuroradiographic partial response; 11 patients (46%), stable disease; and 9 patients (38%), progressive disease following a single cycle of tamoxifen. Time to tumor progression ranged from 3 to 25 months (median, 12 months). Survival ranged from 3 to 27 months (median, 13 months). Five patients are alive, with 3 receiving alternative chemotherapy regimens and 2 continuing to receive tamoxifen. In the group with responding and stable disease, median survival was 15 months (range, 8-27 months). CONCLUSION: Tamoxifen demonstrated modest efficacy with no apparent toxic effects in this heavily pretreated cohort of young adults with recurrent anaplastic astrocytomas. PMID- 10369311 TI - Deletions causing spinal muscular atrophy do not predispose to amyotrophic lateral sclerosis. AB - BACKGROUND: Amyotrophic lateral sclerosis (ALS) is a rapidly progressive, invariably lethal disease resulting from the premature death of motor neurons of the motor cortex, brainstem, and spinal cord. In approximately 15% of familial ALS cases, the copper/zinc superoxide dismutase gene is mutated; a juvenile form of familial ALS has been linked to chromosome 2. No cause has been identified in the remaining familial ALS cases or in sporadic cases and the selective neurodegenerative mechanism remains unknown. Deletions in 2 genes on chromosome 5q, SMN (survival motor neuron gene) and NAIP (neuronal apoptosis inhibitory protein gene), have been identified in spinal muscular atrophy, a disease also characterized by the loss of motor neurons. These genes are implicated in the regulation of apoptosis, a mechanism that may explain the cell loss found in the brains and spinal cords of patients with ALS. OBJECTIVE: To determine whether the mutations causing neurodegeneration in spinal muscular atrophy are present in patients with ALS in whom the copper/zinc superoxide dismutase gene is not mutated. PATIENTS AND METHODS: Patients in whom ALS was diagnosed were screened for mutations in the SMN and NAIP genes by single strand conformation analysis. RESULTS: We found 1 patient with an exon 7 deletion in the SMN gene; review of clinical status confirmed the molecular diagnosis of spinal muscular atrophy. No mutations were found in the remaining patients. CONCLUSION: The SMN and NAIP gene mutations are specific for spinal muscular atrophy and do not predispose individuals to ALS. PMID- 10369312 TI - Neurofibrillary tangles in nondemented elderly subjects and mild Alzheimer disease. AB - BACKGROUND: The relationship between neuropathological lesions and mild, "preclinical," cognitive impairments of Alzheimer disease is poorly understood. Identification of the lesions that are most closely associated with the earliest symptoms of Alzheimer disease is crucial to the understanding of the disease process and the development of treatment strategies to affect its progression. DESIGN AND MAIN OUTCOME MEASURES: We examined the extent of neurofibrillary tangles (NFTs) in 4 neocortical regions, the hippocampus, the entorhinal cortex, and the amygdala in 65 elderly subjects with no dementia, questionable dementia, mild dementia, or moderate dementia as assessed using the Clinical Dementia Rating Scale (CDR). SETTING AND PATIENTS: Postmortem study of nursing home residents. RESULTS: Neurofibrillary tangles were present in the entorhinal cortex and the hippocampus of all subjects, including those without cognitive deficits. Neocortical NFTs were mostly absent in the nondemented (CDR score, 0.0) subjects. The density of NFTs in the questionably demented (CDR score, 0.5) subjects was not significantly increased (P>.20) relative to the nondemented group in any of the brain regions studied. Significant increases (P<.04) in NFT density become apparent first in the amygdala and the temporal cortex in subjects rated to be mildly impaired (CDR score, 1.0). By the time that cognitive impairments were judged to be moderately severe (CDR score, 2.0), all regions of the brain examined, except for the occipital cortex, were significantly (P<.05) involved. CONCLUSIONS: Some NFTs are present in the entorhinal cortex and hippocampus of most elderly individuals irrespective of their cognitive status, but the density of NFTs increases as a function of dementia severity. PMID- 10369313 TI - Familial paroxysmal dystonic choreoathetosis: clinical findings in a large Japanese family and genetic linkage to 2q. AB - BACKGROUND: Paroxysmal dystonic choreoathetosis (PDC) is a rare familial movement disorder that has been mapped to chromosome 2q31-36. OBJECTIVE: To study the first Japanese family with PDC clinically and genetically. PATIENTS AND METHODS: We studied a large Japanese family in which at least 17 members in 6 generations have been affected by PDC. We interviewed and examined 26 family members, 8 of whom revealed choreoathetosis-like and dystonialike involuntary movement and 1 of whom revealed no involuntary movement but only muscle stiffness such as the aura of paroxysmal dystonic choreoathetosis (PDC). Genetic linkage studies of this family were carried out with polymorphic DNA markers. RESULTS: The attacks of involuntary movement or muscle stiffness were precipitated by ovulation, menstruation, emotional stress, or caffeine or alcohol ingestion. Magnetic resonance imaging of the brain revealed no abnormalities. Clonazepam therapy was effective for reducing the attacks, and ingestion of garlic was believed by patients to be effective for softening the attacks. An affected woman with only muscle stiffness showed remission after hysterectomy for hysteromyoma. This woman also had the disease haplotype and transferred it to her typical PDC-affected daughter. Maximal pairwise logarithm of odds scores exceeding 2.00 were obtained at D2S2250, D2S1242, D2S377, D2S2148, and D2S126. The PDC gene was demonstrated by linkage analyses to be located in a 15.3-centimorgan interval lying between D2S371 and D2S339 based on pairwise and multipoint logarithm of odds scores and obligate recombination events in affected individuals. CONCLUSIONS: Linkage of PDC to chromosome 2q32-36 was confirmed in a Japanese family. The clinical characterizations of this family with PDC include that ovulation seems also to be a precipitating factor of the attacks and that hysterectomy seems to be effective for softening the attacks. Although low-dose clonazepam treatment was most effective, garlic use was believed by affected members to be effective for softening the attacks. Furthermore, based on the results of clinical and genetic analyses, we suggest that muscle stiffness without involuntary movement may represent a forme fruste of PDC. PMID- 10369314 TI - Trigeminal neuralgia triggered by auditory stimuli in multiple sclerosis. AB - OBJECTIVES: To describe a patient with a demyelinating brainstem lesion who developed right-sided trigeminal neuralgia triggered by auditory stimuli and to discuss the pathophysiological mechanisms underlying this unusual phenomenon. DESIGN: Case report. SETTING: Referral center. PATIENT: A 27-year-old man who presented with clinical signs of a brainstem lesion developed right-sided trigeminal neuralgia triggered by auditory stimuli to the right ear. Magnetic resonance imaging and electrophysiological studies demonstrated a demyelinating lesion in the pons affecting the right lateral lemniscus and the right trigeminal pathway. This phenomenon completely subsided within 4 days. After a relapse, the diagnosis of clinically definite multiple sclerosis was made. CONCLUSION: Lateral spread of impulse activity within the demyelinating pontine lesion is the likely explanation for the unusual phenomenon of trigeminal neuralgia triggered by auditory stimuli. PMID- 10369316 TI - Do we have a treatment for Alzheimer disease? Yes. PMID- 10369315 TI - Do we have drugs for dementia? No. PMID- 10369317 TI - Seizures in the life and works of Edgar Allan Poe. AB - Edgar Allan Poe, one of the most celebrated of American storytellers, lived through and wrote descriptions of episodic unconsciousness, confusion, and paranoia. These symptoms have been attributed to alcohol or drug abuse but also could represent complex partial seizures, prolonged postictal states, or postictal psychosis. Complex partial seizures were not well described in Poe's time, which could explain a misdiagnosis. Alternatively, he may have suffered from complex partial epilepsy that was complicated or caused by substance abuse. Even today, persons who have epilepsy are mistaken for substance abusers and occasionally are arrested during postictal confusional states. Poe was able to use creative genius and experiences from illness to create memorable tales and poignant poems. PMID- 10369318 TI - Stroke: how large a public health problem, and how can the neurologist help? AB - Stroke is an enormous public health problem, the magnitude of which can be reduced mainly by effective stroke prevention and less so by effective treatment of acute stroke. The greatest effect is likely to be achieved by a mass approach to prevention, which consists of modification of lifestyle behaviors (eg, less smoking and less intake of salt, alcohol, and fat) among the general population through public education and, more importantly, government legislation. The appropriate identification and treatment of high-risk individuals by neurologists is likely to have a smaller but complimentary impact on the population burden of stroke and a substantial impact on the burden of stroke among individuals. The most cost-effective interventions for patients with transient ischemic attack and ischemic stroke are organized multidisciplinary acute care and rehabilitation in a stroke unit and early secondary prevention with aspirin, blood pressure control, smoking cessation, and, in the appropriate patient, oral anticoagulant therapy and carotid endarterectomy. The cost-effectiveness of carotid endarterectomy for asymptomatic carotid stenosis is highly questionable until data from ongoing trials (eg, Asymptomatic Carotid Surgery Trial) become available. Screening for asymptomatic carotid stenosis is more likely to be harmful than helpful, except perhaps among populations with a very high prevalence (pretest probability) of severe carotid stenosis. It is essential that the impact of these strategies on the incidence, outcome, and cost of stroke is measured and monitored. Currently, this is done simply, but unreliably, by examining changes in statistics that are already being measured, such as mortality (eg, among those younger than 70 years old, for greater accuracy). A growing priority in many countries is the development and implementation of valid, reliable, practical, and inexpensive methods of routinely collecting and evaluating data on stroke incidence, outcome, and cost. PMID- 10369319 TI - Anticardiolipin antibodies. PMID- 10369320 TI - The measurement of nitrite in adulterated urine samples by high-performance ion chromatography. AB - With the increased availability of nitrite-containing commercial products that are used for the adulteration of urine samples in the workplace, it is necessary for laboratories to be able to detect and confirm the presence of nitrite in these samples. We have developed a method to confirm the presence of nitrite in urine samples. The method uses the IonPac AS 14 analytical column with the Dionex series 45001 Bio LC system equipped with an anion self-generating suppressor and conductivity detector. Using a single-point calibration, the method is linear and accurately quantitates nitrite to 12,000 microg/mL. The limit of detection is 30 microg/mL, and the day-to-day precision of the assay has a coefficient of variation (CV) of 4.3% at 1200 microg/mL and 3.8% at 2700 microg/mL of nitrite. PMID- 10369321 TI - Prevalence of drugs used in cases of alleged sexual assault. AB - In recent years, there has been an increase in the number of reports in the U.S. of the use of drugs, often in conjunction with alcohol, to commit sexual assault. A study was undertaken to assess the prevalence of drug use in sexual assault cases in which substances are suspected of being involved. Law enforcement agencies, emergency rooms, and rape crisis centers across the U.S. were offered the opportunity to submit urine samples collected from victims of alleged sexual assault, where drug use was suspected, for analysis of alcohol and drugs which may be associated with sexual assault. Each sample was tested by immunoassay for amphetamines, barbiturates, benzodiazepines, cocaine metabolite (benzoylecgonine), cannabinoids, methaqualone, opiates, phencyclidine and propoxyphene. The positive screen results were confirmed by gas chromatography mass spectroscopy (GC-MS). In addition, each sample was tested for flunitrazepam metabolites and gamma-hydroxybutyrate (GHB) by GC-MS and for ethanol by gas chromatography-flame ionization detection (GC-FID). Over a 26-month period, 1179 samples were collected and analyzed from 49 states, Puerto Rico, and the District of Columbia. The states sending the most samples were California (183), Texas (119), Florida (61), Pennsylvania (61), New York (61), Minnesota (50), Illinois (47), Indiana (44), Michigan (40), Maryland (37), Virginia (32), and Massachusetts (31). Four-hundred sixty eight of the samples were found negative for all the substances tested; 451 were positive for ethanol, 218 for cannabinoids, 97 for benzoylecgonine, 97 for benzodiazepines, 51 for amphetamines, 48 for GHB, 25 for opiates, 17 for propoxyphene, and 12 for barbiturates. There were no samples identified as positive for phencyclidine or methaqualone. In addition, 35% of the drug-positive samples contained multiple drugs. This study indicates that, with respect to alleged sexual assault cases, the prevalence of ethanol is very high, followed by cannabinoids, cocaine, benzodiazepines, amphetamines, and GHB. Although only a couple of substances have been implicated with sexual assault, this study has shown that almost 20 different substances have been associated with this crime. This study also raises the concern of illicit and licit drug use in sexual assault cases and suggests the need to test for a range of drugs in these cases. It also highlights the need to test for GHB, which is not generally tested for in a normal toxicology screen. PMID- 10369322 TI - Nail analysis for drugs of abuse: extraction and determination of cannabis in fingernails by RIA and GC-MS. AB - Fingernail clippings were evaluated as analytical specimens for the detection and quantitation of cannabinoids. Specimens were obtained from consenting adults attending a drug clinic, along with information concerning the drugs which they had used over the previous six months. Methods for the surface decontamination and extraction of the specimens were evaluated. Detergent, water, and methanol washes followed by alkaline hydrolysis and liquid-liquid extraction were selected for use in the study. Extracts were analyzed by radioimmunoassay (RIA) and gas chromatography-mass spectrometry (GC-MS) to detect and quantitate cannabinoids present in fingernail clippings. Positive RIA results were obtained from specimens from six known cannabis users. The mean cannabinoid concentration in fingernail clippings determined by RIA was 1.03 ng/mg. Using GC-MS, the mean delta9-tetrahydrocannabinol concentration in fingernail clippings from a further 14 known cannabis users was 1.44 ng/mg. Using GC-MS, the average 11-nor-delta9 tetrahydrocannabinol-9-carboxylic acid concentration in fingernail clippings from three known cannabis users extracted in acidic pH was 19.85 ng/mg. Based on these results, fingernails are potentially useful biological specimens for the detection of past cannabis use in cases of medicolegal interest. PMID- 10369323 TI - Measurement of cadmium-induced metallothionein in urine by ELISA and prevention of overestimation due to polymerization. AB - Urinary metallothionein (MT) is a biological marker of cadmium (Cd) exposure and Cd-induced renal dysfunction. The MT is prone to oxidation due to high cysteine content and forms polymers, which can result in overestimation of the protein by immunochemical methods. The objectives of the present study were to develop an enzyme-linked immunosorbent assay (ELISA) for the measurement of MT in urine and to find ways by which the protein could either be preserved in its monomeric form or converted to this form before analysis to avoid overestimation. Urine specimens analyzed were either from rats repeatedly injected with Cd or from individuals chronically exposed to cadmium through their diets. The MT in rat urine remained in the monomeric form if the urine was collected at 4 degrees C but did not if it was collected at room temperature. The MT was also polymerized if the urine was subjected to repeated freezing and thawing. Overestimation of MT in rat urine occurred (as much as 12-fold) if the MT was polymerized. Addition of 5mM mercaptoethanol to freshly collected rat urine retarded MT polymerization, and addition of 50mM mercaptoethanol converted the polymerized MT to its monomeric form. Analysis of MT in frozen human urine samples revealed that if the urines were not treated with mercaptoethanol, the estimates of MT concentration were up to 11-fold higher than in the treated samples. We conclude that the polymerization of MT in rat and human urines is a serious problem and results in overestimation of the protein by ELISA and that this problem could be overcome by the addition of mercaptoethanol to the urine samples prior to analysis. PMID- 10369324 TI - Melatonin: aeromedical, toxicopharmacological, and analytical aspects. AB - Melatonin, a pineal hormone present in the blood of humans and other species, has a distinct diurnal variation in its biosynthesis and, therefore, in its concentration. This variation has suggested the possibility of a regulatory function in day/night-dependent physiological processes such as sleep and has led scientists to explore the effects of administered melatonin on the modulation of circadian rhythms. For the self-treatment of sleep disorders and other benefits, melatonin use has been extolled to the extent that 20 million new consumers were added to the U.S. retail market in 1995. Its principal aeromedical application has been in the experimental treatment of jet-lag effects. For aircraft passengers, melatonin administration at destination bedtime appears to improve sleep quality and to decrease the time required to reestablish normal circadian rhythms. For international aircrews that travel through multiple time zones without time to adapt to new environments, taking melatonin before arriving home may further impair already disturbed circadian rhythms. Its use to adjust to shiftwork changes by air traffic controllers, aircraft maintenance workers, and support personnel is even more controversial. Limited studies suggest that giving this hormone to shift workers should be done only under controlled conditions and that taking it at the wrong time may actually impair job performance. Because of its possible interaction with certain medications and the changes in its concentrations observed in some clinical conditions, the practitioner must exercise caution during the medical certification of airmen. The variations in the concentration of melatonin can be effectively determined by radioimmunoassay, high-performance liquid chromatography, and gas chromatography-mass spectroscopy analytical techniques. These techniques are capable of measuring the human daytime (10 pg/mL) and nighttime (30-120 pg/mL) melatonin in plasma/serum. Melatonin measurements in victims of accidental death may allow forensic scientists and accident investigators to use the relationship between its concentration and the time of day when death occurred. The most accurate estimations of the time of death result from analysis of melatonin content of the whole pineal body, whereas less accurate estimates are obtained from serum and urine analyses. Pineal levels of melatonin are unlikely to be altered by exogenous melatonin, but its blood and urine levels would change. High blood levels in a daytime crash victim would suggest exogenous supplementation. The possible interfering effects of postmortem biochemical processes on melatonin concentrations in whole blood and in other tissues are not well understood, and there is a need for the continuing research into melatonin's chronobiological properties to define its proper applications and limitations. The indiscriminate use of melatonin by aviation professionals may pose unacceptable safety risks for air travel. PMID- 10369325 TI - Detection of 6-acetylmorphine in vitreous humor and cerebrospinal fluid- comparison with urinary analysis for proving heroin administration in opiate fatalities. AB - The concentrations of morphine and 6-acetylmorphine (6-AM) in urine, cerebrospinal fluid (CSF), and vitreous humor (VH) and the morphine concentrations in blood were determined by gas chromatography-mass spectrometry for 29 fatalities after abuse of heroin either alone or in combination with alcohol and other drugs. 6-AM was found above a quantitation limit of 1 ng/mL in urine in 89% of the cases, in CSF in 68% of the cases, and in VH in 75% of the cases. The 6-AM concentrations in CSF (mean, 10 ng/mL) and VH (mean, 17 ng/mL) were in general much smaller than in urine (mean, 170 ng/mL); therefore, the different pharmacokinetic behavior of the fluids is discussed. There is no uniformity between the three fluids with respect to the presence or absence of 6 AM. Therefore, CSF or VH may be used as complementary or alternative materials to urine in order to prove heroin uptake in opiate fatalities. PMID- 10369326 TI - Procainamide inhibition of human hepatic degradation of cocaine and cocaethylene in vitro. AB - Procainamide (PA), a cardioactive drug, inhibited the degradation of both cocaine (COC) and cocaethylene (CE) when either was incubated in human liver homogenates for 3 h at 37 degrees C. PA appeared to enhance the formation of CE when COC and ethanol (ETOH) were incubated together in liver homogenate. These observations are clinically significant because cardiotoxicity is common after COC abuse and because PA may be administered to individuals who use COC alone and with ETOH. PMID- 10369328 TI - Amphetamine and fenproporex levels following multidose administration of fenproporex. AB - Drugs that are metabolized to amphetamine or methamphetamine are potentially of significant concern in the interpretation of positive drug-testing results for amphetamines. A number of different drugs have been reported to produce amphetamine in the urine of users. One of these compounds, fenproporex, has been shown to be metabolized to amphetamine, and previous reports indicated the parent compound could be detected at low levels for up to 48 h. Administration of fenproporex for seven days (one 10-mg dose per day) to five healthy volunteers resulted in amphetamine being detected in the urine of all subjects. Peak concentrations of amphetamine ranged from approximately 2850 to 4150 ng/mL. Amphetamine could be detected (> or = 5 ng/mL) in the urine for up to nearly 170 h after the last dose. Analysis of the metabolically produced amphetamine showed the presence of both enantiomers, which can be helpful in the differentiation of some illicit amphetamine use from the use of this precursor drug. In addition, evaluation of the enantiomeric composition of the metabolite (amphetamine) can be a valuable tool in the interpretation of time since last dose. More significantly, all samples that contained amphetamine at a concentration of > or = 500 ng/mL were shown to also contain detectable amounts of the parent compound. PMID- 10369327 TI - GC-MS confirmation of codeine, morphine, 6-acetylmorphine, hydrocodone, hydromorphone, oxycodone, and oxymorphone in urine. AB - A procedure for the simultaneous confirmation of codeine, morphine, 6 acetylmorphine, hydrocodone, hydromorphone, oxycodone, and oxymorphone in urine specimens by gas chromatography-mass spectrometry (GC-MS) is described. After the addition of nalorphine and naltrexone as the two internal standards, the urine is hydrolyzed overnight with beta-glucuronidase from E. coli. The urine is adjusted to pH 9 and extracted with 8% trifluoroethanol in methylene dichloride. After evaporating the organic, the residue is sequentially derivatized with 2% methoxyamine in pyridine, then with propionic anhydride. The ketone groups on hydrocodone, hydromorphone, oxycodone, oxymorphone, and naltrexone are converted to their respective methoximes. Available hydroxyl groups on the O3 and O6 positions are converted to propionic esters. After a brief purification step, the extracts are analyzed by GC-MS using full scan electron impact ionization. Nalorphine is used as the internal standard for codeine, morphine, and 6 acetylmorphine; naltrexone is used as the internal standard for the 6-keto opioids. The method is linear to 2000 ng/mL for the 6-keto-opioids and to 5000 ng/mL for the others. The limit of quantitation is 25 ng/mL in hydrolyzed urine. Day-to-day precision at 300 and 1500 ng/mL ranged between 6 and 10.9%. The coefficients of variation for 6-acetylmorphine were 12% at both 30 and 150 ng/mL. A list of 38 other basic drugs or metabolites detected by this method is tabulated. PMID- 10369329 TI - Another use of silica gel and aqueous eluent for HPLC analysis of clozapine and desmethylclozapine. AB - A procedure that involves a high-performance liquid chromatographic system with silica-bonded columns and reversed-phase eluents was fitted from a previously described method to measure clozapine and desmethylclozapine plasma levels. Clozapine and its demethylated metabolite were extracted from alkalinized serum by a liquid-liquid extraction, separated in 10 min, then quantitated at 254 nm at a minimum concentration of 20 ng/mL. The standard curves were linear over the range of 50-3000 ng/mL (r > 0.99) both for clozapine and desmethylclozapine and the assay had good sensitivity and recovery. Intra- and interday coefficients of variation for 200 and 800 ng/mL controls were less than 11.5% for clozapine and desmethylclozapine. This simple and efficient assay was used to monitor clozapine and desmethylclozapine levels from some treatment-refractory schizophrenic patients. PMID- 10369330 TI - Etodolac in equine urine and serum: determination by high-performance liquid chromatography with ultraviolet detection, confirmation, and metabolite identification by atmospheric pressure ionization mass spectrometry. AB - A high-performance liquid chromatographic method was used for the detection of etodolac in equine serum and urine. The method consisted of a one-step liquid liquid extraction, separation on a reversed-phase (RP-18) column and detection using an ultraviolet detector. Additional confirmation methods included a HPLC coupled with an atmospheric pressure chemical ionization mass spectrometer (APCI MS). Free (unbound) etodolac and its conjugates were present in the samples. Concentrations of the drug in the serum and urine samples collected from four standardbred mares after a single oral administration of Ultradol were determined. Maximum etodolac concentrations of 712, 716, 568, and 767 microg/mL in urine and 4.1, 3.6, 3.1, and 2.2 microg/mL in serum were observed. The peak concentrations of the drug were detected 2-10 h (urine) and 40 min-6 h (serum) after administration to four horses. The maximum detection time was 79 h in urine and 48 h in serum after the drug administration. The drug-elimination profiles for both urine and serum are presented and discussed. Method ruggedness and precision and stability studies of etodolac in serum and urine are presented. Three major metabolites were detected in the urine by liquid chromatography-APCI MS. All three metabolites were identified as monohydroxylated etodolac. PMID- 10369331 TI - Pitfalls when determining tissue distributions of organophosphorus chemicals: sodium fluoride accelerates chemical degradation. AB - This paper describes the tissue distributions of dichlorvos, an organophosphate, and chlorpyrifos-methyl, an organophosphorothioate, in a male individual who died after ingesting an insecticidal preparation containing these chemicals and the results of an in vitro stability study on dichlorvos and chlorpyrifos-methyl in blood and buffers. Tiny amounts of dichlorvos, 0.067 and 0.027 mg/L, were detected in the vitreous humor and cerebrospinal fluid, respectively. Although dichlorvos (0.082-8.99 mg/L or mg/kg) was detected in the thoracic aortic blood, thoracic inferior vena caval blood, pericardial fluid, bile, and spleen, it was strongly suggested that it had diffused postmortem from the stomach, which contained 879 mg, because no dichlorvos was detected in the other blood samples and tissues tested. Substantial amounts (0.615-4.15 mg/L) of chlorpyrifos-methyl were detected in all blood samples, and the order of its concentrations was as follows: pulmonary vessel blood > thoracic inferior vena caval blood > blood in the right cardiac chambers > blood in the left cardiac chambers approximately thoracic aortic blood > right femoral venous blood. The total amount of chlorpyrifos-methyl in the stomach was 612 mg. However, it was strongly suggested that virtually no chlorpyrifos-methyl diffused from the stomach into surrounding fluids and tissues postmortem because no chlorpyrifos-methyl was detected in the bile and little was found in the pericardial fluids. Neither compound was detected in the urine. In vitro experiments showed that dichlorvos (10 mg/L) almost disappeared from fresh (pH 7.4) and acidified (pH 6.2) blood samples within 24 and 72 h, respectively. However, 53 and 77% of the original amount of dichlorvos in 0.05M phosphate buffers at pH 7.4 and 6.2 were detected 72 h later. Chlorpyrifos-methyl (1 mg/L) was very stable in blood samples, regardless of the pH, during the 72-h study period, but in the pH 7.4 and 6.2 phosphate buffers, approximately 80% of the original amount had degraded after 72 h. These results indicate that organophosphates are degraded more rapidly by esterase activities than by chemical mechanisms and that organophosphorothioates are hydrolyzed chemically in aqueous solutions but are very stable in biological specimens and not metabolized by esterases. When sodium fluoride was added to blood samples, dichlorvos degraded completely within 15 min, and chlorpyrifos-methyl became very unstable. Thus, when analyzing samples to detect organophosphorus chemicals, this common preservative should not be added to fluid specimens. PMID- 10369332 TI - Comparison of solid-phase extraction and supercritical fluid extraction for the analysis of morphine in whole blood. AB - A comparative study of the quantitative determination of morphine in whole blood using solid-phase extraction (SPE) and supercritical fluid extraction (SFE) is described. Comparative studies were made of the two techniques for the extraction of morphine from authentic forensic blood specimens. Quantitative results indicate that morphine levels measured using SPE correspond well to morphine levels produced using SFE. The two techniques are therefore comparable, although SFE is faster and cleaner and extracts may be produced with higher analyte recoveries than with SPE. This paper presents a comparison of the two techniques and the morphine concentrations determined in blood. PMID- 10369333 TI - Analytical detection and quantitation of strychnine in chemically fixed organ tissues. AB - This study reports the results of the detection and quantitation of strychnine in formalin-fixed tissues and in the formalin solutions in which the tissues were fixed. The toxicological analyses were performed on formalin-fixed liver and kidney samples and formalin solutions (10% buffered pH 7) in which the same samples from a case of acute strychnine poisoning were preserved. The analyses carried out at the time of autopsy on body fluid and tissues (bile, 2.40 mg/L; stomach contents, 14.2 mg; liver, 6.68 mg/kg; kidney, 2.68 mg/kg) allowed the identification of this substance as cause of death. The tissue samples were preserved in formalin solutions for 8 weeks. The analyses performed on formalin fixed tissues (liver and kidney) and on formalin solutions, in which the same tissues were preserved, permitted the detection and quantitation of strychnine (liver, 1.59 mg/kg; formalin from the liver, 1.80 mg/L; kidney, 0.98 mg/kg; formalin from the kidney, 1.11 mg/L). The results indicate that this particular toxic substance also shows good stability in biological specimens subjected to chemical fixation. PMID- 10369334 TI - Delta 9-tetrahydrocannabivarin (delta 9-THCV) as a marker for the ingestion of cannabis versus Marinol. PMID- 10369335 TI - First identification of prednisone in human hair by liquid chromatography ionspray mass spectrometry. PMID- 10369336 TI - More data about the new psychoactive drug 2C-B. PMID- 10369337 TI - Multidetector helical CT angiography: poor cousin or contender? PMID- 10369338 TI - Paradoxical hyperfixation of HMPAO in cerebral infarction. PMID- 10369339 TI - Interventional MR imaging: concepts, systems, and applications in neuroradiology. PMID- 10369340 TI - Paravertebral arteriovenous malformations with epidural drainage: clinical spectrum, imaging features, and results of treatment. AB - BACKGROUND AND PURPOSE: Arteriovenous malformations (AVMs) of the spine or spinal cord can be characterized as spinal cord AVMs, spinal cord and dural arteriovenous fistulas, and AVMs occurring outside the dura but draining into the epidural veins. The purpose of this study was to review the clinical spectrum, imaging features, and results of treatment of paravertebral arteriovenous malformations (PVAVMs) with epidural drainage. METHODS: The clinical records and images of 10 patients with PVAVMs were analyzed retrospectively for clinical presentation, MR findings, angioarchitecture, pathophysiology, treatment efficacy, and clinical follow-up. RESULTS: Seven patients had myelopathy. The MR findings for three of these patients showed spinal cord hyperintensity on T2 weighted sequences and prominent perimedullary vessels. Angiography, performed in two of the three patients, showed evidence of reflux into the perimedullary veins from the PVAVMs. Each of these two patients underwent surgical clipping of the radicular vein leading to the perimedullary veins. In three of the seven patients, there were large epidural veins compressing the cord. Angiography performed in these patients showed large PVAVMs with multiple feeders, which were treated by a combination of transarterial and transvenous embolization. One of the seven patients had an associated spinal cord arteriovenous malformation. In three patients with incidental PVAVMs, cure was achieved by using a combination of coils and liquid adhesives by the endovascular route. CONCLUSION: The clinical presentation of PVAVMs is variable, and symptomatic lesions are the result of compression by epidural veins or of congestive myelopathy. A clear understanding of the anatomy and pathophysiology is necessary to plan treatment. Endovascular techniques are capable of curing the malformation, alleviating the symptoms, or both in a significant proportion of these lesions. PMID- 10369341 TI - Hypervascular spinal tumors: influence of the embolization technique on perioperative hemorrhage. AB - BACKGROUND AND PURPOSE: Corporectomy is an effective treatment for vertebral metastases; however, massive perioperative hemorrhage is often associated with this procedure. We compared preoperative particle, particle-coil, and coil embolizations of hypervascular spinal tumors prior to vertebral body replacement to determine which prevented perioperative hemorrhage most effectively. METHODS: The vertebral tumors of 59 patients were embolized prior to corporectomy. In 26 cases, only coils were used for the proximal occlusion of feeding segmental arteries. Twenty-four patients received a combination of polyvinyl alcohol (PVA) particles and coils, and nine tumors were embolized with particles alone. We compared intraoperative blood loss between the three groups and 10 other patients who did not undergo embolization prior to corporectomy. RESULTS: Estimation of intraoperative hemorrhage showed a median value of 4350 mL in patients without embolization, 2650 mL in cases of coil embolization, 1850 mL in cases of particle coil embolization, and 1800 mL in cases of particle embolization. The difference between unembolized patients and those who underwent coil embolization was not statistically significant. Particle and particle-coil embolizations showed very similar results, and reduced hemorrhage significantly as compared to unembolized and proximal coil occlusion cases. Residual bleeding came from the venous system and the neighborhood of the embolized region. CONCLUSION: Particle embolization prior to corporectomy can reduce perioperative hemorrhage. The additional benefit of proximal coil occlusion of arterial feeders is questionable. PMID- 10369342 TI - A simplified arteriovenous malformation model in sheep: feasibility study. AB - BACKGROUND AND PURPOSE: Recently, a swine model of a cerebral arteriovenous malformation (AVM) has been developed that closely resembles a human AVM of the brain. The creation of such a model requires sophisticated neurointerventional techniques. The purpose of this study was to develop a simple and cost-effective AVM animal model that does not require additional endovascular techniques. METHODS: A surgical anastomosis was created in seven sheep between the common carotid artery and the ipsilateral jugular vein, followed by ligation of the jugular vein above the anastomosis and of the proximal common carotid artery below the anastomosis. The anastomosis was created on the left side in four animals and on the right side in three. Cerebral angiography from the contralateral carotid artery was performed before and immediately after surgery to delineate the relevant cerebral vascular anatomy and to determine the direction of blood flow. RESULTS: An angiographic appearance simulating an AVM was found in all the animals. The ramus anastomoticus and arteria anastomotica functioned as the feeding vessels to the rete mirabile, which represented the nidus in our model, and to the jugular vein, which represented the draining vein from the malformation. Extensive collateral flow through the rete mirabile into the distal segment of the external carotid artery above the ligature was observed angiographically, with retrograde flow through the surgical anastomosis into the jugular vein. CONCLUSION: A simple surgically created experimental model for cerebral AVMs was developed in sheep without the need for additional complex endovascular catheter manipulations of intracranial branches. Such an animal model can substantially reduce the cost of research and training in the neurointerventional or radiosurgical management of AVMs. PMID- 10369343 TI - Cavernous aneurysm rupture with balloon occlusion of a direct carotid cavernous fistula: postmortem examination. AB - We present a unique case of a patient with a symptomatic carotid cavernous fistula treated successfully with balloon embolization. Her subsequent death from other disease processes allowed direct visualization of the balloon occlusion in situ at postmortem examination. PMID- 10369344 TI - The porous, guidewire-directed, detachable aneurysm liner: a new concept in the endovascular treatment of intracranial aneurysms. AB - A new device for the endovascular treatment of aneurysms is described. It consists of a guidewire-directed porous liner or bag, detachably mounted on a microcatheter and designed to be inserted into an aneurysm and to be filled with detachable coils or other embolic agents. Several prototypes have been made. Preliminary in vitro and in vivo experiments have demonstrated its behavior in relatively wide-necked aneurysms. PMID- 10369345 TI - Treatment of distal aneurysms of the cerebellar arteries by intraaneurysmal injection of glue. AB - Distal aneurysms of the cerebellar arteries are associated with a poor prognosis, as surgery or embolization with GDCs is very difficult. We report our experience with a new therapeutic method involving intraaneurysmal injection of glue. Three aneurysms were catheterized with a flow-guided microcatheter, and glue was slowly injected into the aneurysms. In two cases, treatment resulted in total occlusion of the aneurysm with preservation of the parent artery. In one case, the aim was to occlude both the aneurysm and parent artery. PMID- 10369346 TI - Cerebrovascular reserve before and after vertebral artery angioplasty. AB - The selection of patients with severe vertebrobasilar artery stenosis for angioplasty is based mainly on clinical experience rather than on controlled data. We present a patient with severe vertebral artery stenosis in whom we could document the positive effect of angioplasty on posterior circulation hemodynamics by using transcranial Doppler sonography. PMID- 10369347 TI - Cerebral angioplasty and stenting for intracranial vertebral atherosclerotic stenosis. AB - A 72-year-old man underwent cerebral angioplasty and stenting for a high-grade eccentric atherosclerotic stenosis (93%) of the right intracranial vertebral artery. The lesion was sufficiently and smoothly dilated very easily with the use of a highly flexible, balloon-expandable coronary stent. No complications occurred during or after the procedure. This therapeutic option may prove to be a safe and useful means to resolve an intracranial atherosclerotic stenosis. PMID- 10369348 TI - Intracranial stenoocclusive disease: double-detector helical CT angiography versus digital subtraction angiography. AB - BACKGROUND AND PURPOSE: To our knowledge, no large-scale studies comparing the accuracy of CT angiography (CTA) to intraarterial digital subtraction angiography (DSA) of intracranial stenosis have been reported. We attempted to determine the diagnostic value of intracranial CT angiography (CTA) of normal vasculature and variants as well as of stenoocclusive disease. METHODS: One-hundred and twelve patients underwent CTA and intraarterial angiography, and 2205 vascular segments were examined to ascertain presence, visibility, and degree of arterial stenoses (n = 105) as well as anatomic variants. Source, maximum intensity projection (MIP), and MIP-generated multiplanar reformatted (MPR) images were evaluated. RESULTS: All 55 anatomic variants were identified correctly. Visibility of small vessel segments was increased from 75% to 83% by using source images. MPR was helpful in differentiating distal vertebral hypoplasia from stenosis and in overcoming artifacts. All 43 occlusive segments were graded correctly (sensitivity = 100%, predictive value = 93.4%) as follows: severely stenotic ([n = 23], sensitivity = 78%, predictive value = 81.8%); moderately stenotic ([n = 36], sensitivity = 61%, predictive value = 84.6%); and mildly stenotic ([n = 3], sensitivity = 66%, predictive value = 28%). Normal segments (n = 2100) had a sensitivity of 99.5%, and CTA evinced a specificity of 99% for detecting stenoocclusive disease. Approximately one-third of wrong assessments were related to the petrous segment of the carotid artery. CONCLUSION: CTA with double detector technology and advanced postprocessing algorithms, including MPR, is about as reliable as MRA in depicting the vasculature of the anterior and posterior circulation and in grading intracranial stenoocclusive lesions, with the exception of the petrous segment of the carotid artery. CTA might be superior to MRA in the evaluation of poststenotic low-flow segments. PMID- 10369349 TI - Digitized cerebral synchrotron radiation angiography: quantitative evaluation of the canine circle of Willis and its large and small branches. AB - BACKGROUND AND PURPOSE: Conventional X-ray angiography lacks the sensitivity and spatial resolution needed to detect small amounts of iodinated contrast material and to quantitate diameters of the small vessels in the brain. The purpose of this study was to ascertain whether digitized synchrotron radiation microangiography, with the use of a high-definition TV camera system, can accurately show small cerebral vessels. METHODS: Six anesthetized dogs were exposed to monochromatic synchrotron radiation with an energy level of 33.3 keV optimized for iodine detection while iodinated contrast material was injected into the brachiocephalic and vertebral arteries. The images were detected with a high-definition TV camera system with a spatial resolution of 30 microm. In all, 26 cerebral angiograms of the circle of Willis with its branches were obtained, and the images were digitized at a workstation. RESULTS: The small branches of the circle of Willis were clearly visible on all images. Vasodilatation of the circle of Willis and its large and small branches induced by CO2 inhalation was quantitatively confirmed on the images: for example, the diameter of one small branch was increased from 0.24 +/- 0.04 mm to 0.38 +/- 0.12 mm. Temporal subtraction improved the image quality. CONCLUSION: The synchrotron radiation angiographic system is useful for visualizing large and small vessels deep in the brain as well as for quantitating their diameters. PMID- 10369350 TI - Tagged MR imaging of intracranial aneurysm models. AB - We used tagged MR imaging to investigate the flow in two lateral and two terminal saccular intracranial aneurysm models of different neck sizes. Imaging was performed with a 1.5-T superconducting MR system using 2D fast spatial modulation of magnetization (SPAMM) sequences with an intersegmental delay of 25 milliseconds. The flow in the saccular aneurysm models varied with the shape and size of the neck: flow was faster in wider-necked aneurysms than in those with narrower necks. PMID- 10369351 TI - Relevance of hypointense lesions on fast fluid-attenuated inversion recovery MR images as a marker of disease severity in cases of multiple sclerosis. AB - BACKGROUND AND PURPOSE: Hypointense lesions can be visible on fast fluid attenuated inversion recovery (FLAIR) MR images of the brain of patients with multiple sclerosis (MS), and they may be produced by severely damaged white matter. To test the role of these lesions as an MR marker of MS severity, we assessed their relationship with clinical findings and other MR measures. METHODS: Using a 1.5-T scanner, dual-echo rapid acquisition with relaxation enhancement, fast FLAIR, and T1-weighted MR images (24 axial, 5-mm-thick contiguous interleaved sections) were obtained from 50 patients (32 with relapsing-remitting and 18 with secondary progressive MS). RESULTS: Hypointense lesions were visible on the fast FLAIR images of 19 patients (mean number of lesions, 7.8; range 1-22); their median load was 1.4 mL (range, 0.05-12.6 mL). The median lesion load was significantly higher in patients with secondary progressive MS than in those with relapsing-remitting MS on the T1-weighted images. Both the number and the load of hypointense lesions shown by fast FLAIR imaging were significantly higher in patients with secondary progressive MS. Significant correlations were found between Expanded Disability Status Scale scores and MR lesion load. A multivariate analysis showed that only the presence of hypointense lesions on fast FLAIR images significantly separated cases of relapsing-remitting MS from cases of secondary progressive MS (relative risk, 7.1; 95% confidence interval, 2.0-25.9). CONCLUSION: The presence of hypointense lesions on fast FLAIR images was a strong predictor of disease severity in cases of MS, although the low sensitivity of this approach might limit its use for the assessment of MS evolution. PMID- 10369352 TI - Long-term changes of magnetization transfer-derived measures from patients with relapsing-remitting and secondary progressive multiple sclerosis. AB - BACKGROUND AND PURPOSE: For cases of multiple sclerosis (MS), magnetization transfer (MT) imaging may provide more pathologically specific and accurate estimates of the disease process than does conventional imaging. In this study, we evaluated changes of the MT ratio (MTR) of newly enhancing lesions, the MTR of normal-appearing white matter (NAWM), the average lesion MTR, and the MT histogram-derived metrics during a 3-year follow-up period for patients with relapsing-remitting or secondary progressive MS. METHODS: Dual-echo, conventional spin-echo, and MT images were obtained from seven patients with relapsing remitting MS, seven patients with secondary progressive MS, and five age- and sex matched control subjects at the time of study entry and 1, 13, and 37 months later. RESULTS: Newly enhancing lesions in the patients with secondary progressive MS presented a more severe and significant MTR reduction during the follow-up period as compared with those in the relapsing-remitting group. In cases of secondary progressive MS, we also observed a significant reduction of the MTR values of the NAWM and a trend toward reduction of average lesion MTR values. The patients with MS had mean percentage changes of MT histogram-derived measures that were approximately two to 10 times higher than those of the control subjects. CONCLUSION: This preliminary 3-year follow-up study shows that newly enhancing lesions and NAWM in patients with secondary progressive MS have significantly lower MTR values than do those in patients with relapsing-remitting MS. It also shows that the tissue damage that remains after enhancement ceases is more severe in secondary progressive disease. PMID- 10369353 TI - Combined magnetization transfer and proton spectroscopic imaging in the assessment of pathologic brain lesions in multiple sclerosis. AB - BACKGROUND AND PURPOSE: Conventional MR imaging of multiple sclerosis (MS) provides relatively poor pathologic specificity, which has led to the investigation of more sophisticated MR techniques. The purpose of this study was to combine magnetization transfer (MT) imaging and proton MR spectroscopic imaging (MRSI) to evaluate the specific pathologic features of myelination and neuronal integrity in patients with MS and to determine the relationship between these measures within plaques. METHODS: We acquired conventional MR, MT, and proton MRSI data and evaluated clinical disability in 30 patients with MS, whose conditions were categorized as relapsing-remitting, primary progressive, or secondary progressive. The lesions were classified, using a semiautomated edge following technique, on T2-weighted MR images, and an analysis of MT and proton MRSI data was conducted for lesion regions as well as for tissue that was categorized as normal. RESULTS: The MT ratio (MTR) of normal-appearing white matter in the patients with MS was significantly lower than in the healthy participants, whereas gray matter values were unchanged. MS lesions showed a large reduction in MTR, with old lesions exhibiting a lower MTR than new lesions. The average lesion MTR and the MR spectroscopic imaging-measured relative concentration of N-acetylaspartate, a marker of neuronal integrity, was positively correlated in patients with relapsing-remitting MS. This relationship was strengthened in regions containing new lesions. CONCLUSION: The integrated use of MT and MR spectroscopic imaging provides a more complete description of the pathologic features of MS than does conventional MR imaging alone, and our data suggest that axonal damage occurs in step with new demyelination and is not a late feature of the disease. PMID- 10369355 TI - The evolution of cerebral blood flow in the developing brain: evaluation with iodine-123 iodoamphetamine SPECT and correlation with MR imaging. AB - BACKGROUND AND PURPOSE: Although it is well established that brain maturation correlates temporally with the functions the newborn or infant performs at various stages of development, the precise relationship between function and anatomic brain maturation remains unclear. The purpose of this study was to investigate the developmental changes of regional cerebral blood flow (rCBF) in infants and children using iodine-123 iodoamphetamine (123I-IMP) and single photon emission computed tomography (SPECT). These findings were correlated with the MR imaging appearance of the brain and with known developmental changes. METHODS: Twenty-one 123I-IMP SPECT examinations of 17 patients, ranging in age from neonates to 2 years, were reviewed retrospectively. All children had had transient neurologic events in the neonatal period that did not significantly affect subsequent neuropsychological development. MR studies were performed in 12 of these patients and the MR findings were correlated with the SPECT results. RESULTS: SPECT studies showed a consistent pattern of evolving changes in 123I IMP uptake, most likely reflecting evolution of rCBF. From the 34th postconceptional week until the end of the second month after term delivery, there was predominant uptake in the thalami, brain stem, and paleocerebellum, with relatively less cortical activity. Radionuclide uptake in both the perirolandic and occipital cortices was well seen around the 40th postconceptional week and increased rapidly thereafter, with a predominance of parietal activity. By 3 months, radionuclide uptake in the cerebellar hemispheres and parietofrontal cortices increased. Frontal and temporal activity increased by age 6 to 8 months. Uptake in the basal ganglia increased by 8 months. By the beginning of the second year, rCBF showed a similar topographic pattern to that in adults. CONCLUSION: The time course of the changes in 123I-IMP uptake in the developing brain as detected by SPECT is similar to that of myelination and most likely reflects an overall topologic maturational pattern of the brain. PMID- 10369354 TI - Magnetization transfer ratio of white matter hyperintensities in subcortical ischemic vascular dementia. AB - BACKGROUND AND PURPOSE: In subjects with subcortical ischemic vascular dementia (SIVD), tissue vacuolization, myelin pallor, and demyelination have been found on pathologic examination of white matter signal hyperintensities (WMSH). Magnetization transfer ratio (MTR) values provide a potential measure of compromised white matter integrity. The purpose of this study was to determine if there were differences in MTR of WMSH between subjects with SIVD and cognitively normal healthy control subjects. METHODS: Fifteen subjects with SIVD and 16 control subjects of comparable age and sex were studied. MTR images were coregistered to MR images segmented into tissue classes (gray matter, white matter, CSF, WMSH, and lacunar infarcts). MTR of WMSH was compared across groups and examined by WMSH location, size, and total burden. RESULTS: WMSH burden was greater in SIVD patients than in control subjects (2.4% vs 0.67%). MTR of WMSH did not differ between groups, but MTR of periventricular WMSH was lower in SIVD patients than in control subjects (37.6% vs 39.4%). Even after accounting for covariant effects of lesion burden, there was still a trend toward reduced periventricular WMSH MTR in the group with dementia. There was no correlation between WMSH MTR and WMSH lesion size. CONCLUSION: These findings are consistent with observations that pathologic changes in vascular dementia are most severe in the periventricular white matter and suggest that insight into the pathophysiology of SIVD might be gleaned from studies of the periventricular region. PMID- 10369356 TI - Brain perfusion imaging in asymptomatic patients receiving cyclosporin. AB - BACKGROUND AND PURPOSE: Cyclosporin has neurotoxic effects in a significant number of transplant patients that are associated with characteristic findings on MR images. Focal abnormalities in cerebral perfusion have been implicated in the pathophysiology of cyclosporin neurotoxicity. In the clinically asymptomatic patient, however, it is not known whether any imaging evidence of cyclosporin's effect on the brain is demonstrable. Our hypothesis was that conventional MR imaging, perfusion MR imaging, and single-photon emission CT (SPECT) could enable detection of subclinical lesions in asymptomatic patients. The ability to detect such lesions might aid in the identification of persons most at risk for clinical neurotoxicity. METHODS: Ten posttransplant patients being treated with cyclosporin were recruited prospectively. Imaging studies were performed within 3 weeks of transplantation. Patients were examined with MR imaging, using standard spin-echo and dynamic contrast-enhanced perfusion techniques, and SPECT scanning. Postprocessing of MR perfusion data was performed to obtain pixel-by-pixel maps of regional cerebral blood volume, peak height, and time-to-peak parameters. RESULTS: The mean age of the patients was 45 +/- 11 years. At the time of imaging, three patients had minor neurologic manifestations commonly associated with cyclosporin (ie, mild tremor, headache), but no patient had clinical neurotoxicity. Findings on conventional MR images, MR perfusion maps, and SPECT perfusion scans were normal in all patients. CONCLUSION: Conventional MR imaging, dynamic perfusion MR imaging, and SPECT do not depict any lesions in asymptomatic patients on cyclosporin. Therefore, it may not be possible for imaging methods to aid in the identification of patients at risk for neurotoxicity. Our findings support previously published conclusions that the lesions visible in patients with clinical neurotoxicity are due to cyclosporin effects and not to preexisting coincidental abnormalities. PMID- 10369357 TI - Neuromagnetic assessment of pathophysiologic brain activity induced by minor head trauma. AB - BACKGROUND AND PURPOSE: Patients with mild traumatic brain injury (TBI) often show significant neuropsychological dysfunction despite the absence of abnormalities on traditional neuroradiologic examinations or EEG. Our objective was to determine if magnetic source imaging (MSI), using a combination of MR imaging and magnetoencephalography (MEG), is more sensitive than EEG and MR imaging in providing objective evidence of minor brain injury. METHODS: Four subject groups were evaluated with MR, MSI, and EEG. Group A consisted of 20 neurologically normal control subjects without histories of head trauma. Group B consisted of 10 subjects with histories of mild head trauma but complete recovery. Group C consisted of 20 subjects with histories of mild head injury and persistent postconcussive symptoms. The 15 subjects included in group D underwent repeat examinations at an interval of 2 to 4 months. RESULTS: No MR abnormalities were seen in the normal control group or the asymptomatic group, but five (20%) of the patients with persistent postconcussive symptoms had abnormal MR findings. EEG was abnormal for one subject (5%) from the normal control group, one (10%) from the asymptomatic group, and five (20%) from the group with persistent postconcussive symptoms. MSI was abnormal for one subject (5%) from the normal control group, one (10%) from the asymptomatic group, and 13 (65%) from the group with persistent postconcussive symptoms. There was a direct correlation between symptom resolution and MSI findings for the symptomatic head trauma group. CONCLUSION: MSI indicated brain dysfunction in significantly more patients with postconcussive symptoms than either EEG or MR imaging (P < .01). The presence of excessive abnormal low-frequency magnetic activity provides objective evidence of brain injury in patients with postconcussive syndromes and correlates well with the degree of symptomatic recovery. PMID- 10369358 TI - Magnetization transfer MR imaging in CNS tuberculosis. AB - BACKGROUND AND PURPOSE: CNS tuberculosis may simulate other granulomas and meningitis on MR images. The purpose of this study was to improve the characterization of lesions in CNS tuberculosis and to assess the disease load using magnetization transfer (MT) imaging. METHODS: A total of 107 tuberculomas in seven patients with or without meningitis and 15 patients with tuberculosis meningitis alone were studied. Fifteen patients with cysticercus granulomas with T2 hypointensity, five patients each with viral and pyogenic meningitis, and two patients with cryptococcal meningitis were also studied. The MT ratios were calculated from tuberculomas, cysticercus granulomas, and thickened meninges in tuberculous, viral, pyogenic, and cryptococcal meningitis and were compared within each pathologic group and with the MT ratio of different regions of normal brain parenchyma. Detectability of lesions on T1-weighted MT spin-echo (SE) images was compared with that on conventional SE and postcontrast MT-SE images. RESULTS: Thickened meninges appeared hyperintense relative to surrounding brain parenchyma in the basal and supratentorial cisterns on precontrast MT-SE images in all 18 patients with tuberculosis meningitis. These meninges were not seen or were barely visible on conventional SE images, and enhanced on postcontrast MT-SE images. The MT ratio from the thickened meninges of tuberculous meningitis was significantly lower than that from the meninges in cryptococcal and pyogenic disease and significantly higher than the meninges in viral meningoencephalitis. The MT ratio from T2 visible and invisible tuberculomas appeared to be significantly lower than that of normal white matter. The MT ratio of T2 hypointense cysticercus granuloma was significantly higher than that of T2 hypointense tuberculoma. CONCLUSION: Precontrast MT-SE imaging helps to better assess the disease load in CNS tuberculosis by improving the detectability of the lesions. With the use of MT ratios, it may be possible to differentiate tuberculosis from similar-appearing infective lesions on MR images. PMID- 10369359 TI - MR spectroscopy of bilateral thalamic gliomas. AB - This study reports the MR spectroscopic patterns of two patients with bithalamic glioma. In one patient, phosphorus (31P) MR spectroscopy was performed. In both patients, the proton MR spectroscopic scans showed an increased creatine phosphocreatine peak in the tumor. In the patient who underwent 31P-MR spectroscopy, an increased phosphocreatine peak was also observed. This group of thalamic tumors may be distinguished from other gliomas clinically, radiologically, and metabolically. PMID- 10369360 TI - Intraventricular meningiomas: MR imaging and MR spectroscopic findings in two cases. AB - CT, MR imaging, MR spectroscopy, and angiography were performed in two men (ages 21 and 48, respectively) with intraventricular meningioma. In both cases, CT and MR imaging showed large tumors located in the trigone of the right lateral ventricle that enhanced intensely after contrast administration. MR spectroscopy was helpful in supporting a preoperative diagnosis of meningioma in both cases. PMID- 10369361 TI - Dural arteriovenous fistula of the cavernous sinus with venous congestion of the brain stem: report of two cases. AB - We present two cases of dural arteriovenous fistula of the cavernous sinus with venous congestion of the brain stem. Both cases were detected by MR imaging and showed significant improvement on MR images after transvenous embolization. PMID- 10369362 TI - High-resolution MR cisternography of the cerebellopontine angle: 2D versus 3D fast spin-echo sequences. AB - BACKGROUND AND PURPOSE: The clinical usefulness of MR cisternography of the cerebellopontine angle, applying 2D or 3D fast spin-echo sequences, has been reported recently. Our purpose was to investigate the cause of signal loss in CSF in the prepontine or cerebellopontine angle cistern on 2D FSE MR images and to compare the cisternographic effects of 2D and 3D FSE sequences. METHODS: Preliminary experiments were performed in four volunteers to assess the causes of signal loss. Initially, using a 2D cardiac-gated cine phase-contrast method with a velocity encoding value of 6 cm/s, we measured the velocity and flow pattern of CSF. Comparisons were made to assess the effects of intravoxel dephasing, amplitude of the section-selecting gradient, echo time (TE), and section thickness. Four healthy subjects and 13 patients with ear symptoms were examined, and multisection 3-mm-thick 2D images and 30-mm-slab, 1-mm-section 3D images were compared qualitatively and quantitatively. Then, 3D MR cisternography was performed in 400 patients with ear symptoms, and qualitative evaluation was performed. RESULTS: In volunteers, the average peak velocity of CSF was 1.2 cm/s. With TE = 250, CSF may move an average of 3 mm, and can be washed out of a 3-mm thick 2D section volume. The CSF signal relative to that of a water phantom decreased gradually as TE increased on single-section 3-mm-thick 2D images. The CSF signal relative to that of the water phantom increased gradually as section thickness increased. No significant differences were noted in intravoxel dephasing and amplitude of the section-selecting gradient. The contrast-to-noise ratio (CNR) between CSF and the cerebellar peduncle, and the visibility of the cranial nerves and vertebrobasilar artery were significantly improved on 3D images in 17 subjects. In images from 400 patients, no significant signal loss in the cistern was observed using 3D FSE. CONCLUSION: CSF signal loss in thin section 2D MR cisternography is mainly attributable to the wash-out phenomenon. 3D acquisition can reduce this phenomenon and provide thinner sections. The scan time for 3D acquisition is not excessive when a long echo train length and half Fourier imaging are used. MR cisternography should be performed using a 3D acquisition. PMID- 10369363 TI - Atherosclerotic plaque at the carotid bifurcation: CT angiographic appearance with histopathologic correlation. AB - BACKGROUND AND PURPOSE: The likelihood that carotid plaque will give rise to cerebral ischemia probably relates to the degree of arterial stenosis and to plaque morphology. The aim of this study was to assess whether features seen at CT angiography might be used to predict carotid plaque stability by comparing CT angiograms with histopathologic examinations of the carotid artery bifurcation. METHODS: Nine patients with symptomatic severe carotid stenosis at intraarterial angiography had CT angiography of the carotid bifurcation before carotid endarterectomy. After endarterectomy, multiple sections of the specimens through the carotid bifurcation were examined histologically. Plaque characteristics recorded included the proportion of necrotic/lipid core, presence of hemorrhage, extent of fibrosis, ulceration, calcification, inflammatory cell infiltrate, and fibrous cap thickness. Corresponding CT angiograms were assessed for plaque size, distribution, and radiodensity as well as presence of calcific density and ulceration. Histologic findings and CT angiograms were compared. RESULTS: Plaque with a large necrotic/lipid core, which was often hemorrhagic, was found in 16 of 23 sections, and in 15 of these this histologic appearance corresponded with patchy or homogeneous low density on CT angiograms. Six of seven predominantly fibrous plaques were of soft-tissue density on CT angiograms. High density consistent with calcification was seen more frequently on CT angiograms than it was detected histologically, but CT angiography depicted plaque ulceration poorly (four ulcers at histology; two false-positive and two false-negative findings at CT angiography). CONCLUSION: CT angiography is a promising method for assessing the lumen and wall of the carotid artery. The apparent correlation between histologic appearance and plaque density on CT angiograms has important implications for the prediction of plaque stability, even though ulceration is shown inconsistently. PMID- 10369364 TI - SAPHO syndrome of the temporomandibular joint associated with sudden deafness. AB - We report a case of arthritis of the temporomandibular joint (TMJ) associated with sclerosing osteomyelitis of the mandible and temporal bone, causing deafness. The presence of a palmoplantar pustulosis established the diagnosis of SAPHO syndrome. SAPHO (an acronym referring to synovitis, acne, palmoplantar pustulosis, hyperostosis, and osteitis) syndrome is defined by the association of characteristic osteoarticular and dermatologic manifestations, with diffuse sclerosing osteomyelitis of the mandible being a part of this entity. We review the literature of SAPHO syndrome with mandibular manifestations and discuss the mechanisms of inflammatory spread from the TMJ to the cochlea. To our knowledge, this is the first description of skull base involvement in a patient with SAPHO syndrome leading to sudden deafness. PMID- 10369365 TI - MR imaging of tuberous sclerosis in neonates and young infants. AB - BACKGROUND AND PURPOSE: The MR imaging appearance of intracranial manifestations in tuberous sclerosis varies with age. The aim of this study was to specify MR characteristics in a coherent group of neonates and infants in order to distinguish them from the mature pattern. METHODS: The MR studies of seven patients under 3 months old were reviewed retrospectively. Imaging appearance, number, and distribution of tubers, white matter anomalies, subependymal nodules, and subependymal giant cell astrocytomas were analyzed. RESULTS: All patients had more white matter anomalies, subependymal nodules, subependymal giant cell astrocytomas, transmantle dysplasias, and left-hemispheric and temporal lesions, but less cortical tubers than did older patients in previous series. The lesions were easy to detect as hyperintense foci on T1-weighted images. Visibility as hypointensities on T2-weighted images was comparatively poor. CONCLUSION: The nodular subependymal and linear parenchymal tuberous sclerosis lesions in infants under 3 months old are hyperintense on T1-weighted images and hypointense on T2 weighted images as opposed to the reverse pattern of signal intensity in older persons. The scarce myelination helps to identify white matter anomalies, which become less visible as myelination progresses. Conversely, purely intracortical tubers are more difficult to diagnose in infants. Because the overall number and conspicuity of all other lesions in our series were greater than in previous series with older subjects, our findings indicate that infant age does not compromise, but facilitates, the correct MR diagnosis of tuberous sclerosis. Therefore, if tuberous sclerosis is clinically suspected within the first 3 months of life, imaging should not be delayed. PMID- 10369366 TI - Developmental changes of cerebral blood flow and oxygen metabolism in children. AB - BACKGROUND AND PURPOSE: Normal values for cerebral blood flow (CBF) and metabolism in adults are well established, but not for children. Our goal, therefore, was to clarify functional developmental changes of the brain in children in relation to CBF and oxygen metabolism. METHODS: We measured regional CBF (rCBF), regional cerebral metabolic rate for oxygen (rCMRO2), and regional oxygen extraction fraction (rOEF), using positron emission tomography (PET). We performed 30 PET studies in 24 children ages 10 days to 16 years (nine boys, 15 girls), using a steady inhalation method with C(15)O2, (15)O2, and 15CO in order to measure rCBF, rCMRO2, and rOEF, respectively. Regions of interest were set in the primary cerebral areas (sensorimotor, visual, temporal, and parietal cortex), cerebral association areas (frontal and visual association), basal ganglia (lenticular and thalamus), and posterior fossa (brain stem and cerebellar cortex). Subjects were grouped by age (< 1, 1 to < 3, 3 to < 8, and > or = 8 years), and the absolute values of the parameters were compared with those obtained from 10 healthy adults. RESULTS: rCBF and rCMRO2 were lower in the neonatal period than in older children and adults, and increased significantly during early childhood. rCBF was higher as compared with adults, peaking around age 7, whereas rCMRO2 was relatively high, with the last area to increase being the frontal association cortex. Both rCBF and rCMRO2 reached adult values during adolescence. No difference in rCBF was observed between the basal ganglia and the primary cerebral cortex; however, it was prominent in the occipital lobe in every age bracket. No significant changes in rOEF were found during childhood. CONCLUSION: The dynamic changes of rCBF and rCMRO2 observed in children probably reflect the physiologic developmental state within anatomic areas of the brain. PMID- 10369367 TI - Infantile fibromatosis of the neck with intracranial involvement: MR and CT findings. AB - CT and MR imaging studies were performed in a 3-year-old boy with infantile fibromatosis arising from the infratemporal fossa and extending into the middle cranial fossa. On CT scans, the lesion was hyperattenuating (44-49 Hounsfield units [HU]), enhancing significantly after application of contrast material (63 66 HU). The MR images showed a multilobulated lesion of heterogeneous signal intensity. The tumor was markedly hypointense on T2-weighted images and slightly hypointense on T1-weighted images relative to brain tissue, iso- or slightly hyperintense relative to tongue muscle on both T2- and T1-weighted images, and enhanced strongly after administration of gadopentetate dimeglumine. PMID- 10369368 TI - The relationship between the functional abilities of patients with cervical spinal cord injury and the severity of damage revealed by MR imaging. AB - BACKGROUND AND PURPOSE: The appearance of the damaged spinal cord after injury correlates with initial neurologic deficit, as determined by the American Spinal Injury Association grade and manual muscle test score, as well as with recovery, as assessed by manual muscle test scores. The purpose of this study was to determine whether the presence of spinal cord hemorrhage and the size and location of spinal cord edema on MR images is predictive of functional recovery in survivors of cervical spinal cord injury (SCI). METHODS: The degree of damage to the cervical spinal cord was measured on the MR images of 49 patients who underwent imaging within 72 hours of sustaining SCI. The effects of hemorrhage and length/location of edema on changes in the value of the motor scale of the functional independence measure (FIM) were assessed on admission to and discharge from rehabilitation. RESULTS: Patients without spinal cord hemorrhage had significant improvement in self-care and mobility scores compared with patients with hemorrhage. There was no significant effect of spinal cord hemorrhage on changes in locomotion and sphincter control scores. The rostral limit of edema positively correlated with admission and discharge self-care scores and with admission mobility and locomotion scores. Edema length had a negative correlation with all FIM scales at admission and discharge. CONCLUSION: The imaging characteristics of cervical SCI (hemorrhage and edema) are related to levels of physical recovery as determined by the FIM scale. Imaging factors that correlate with poor functional recovery are hemorrhage, long segments of edema, and high cervical locations. PMID- 10369369 TI - Idiopathic spinal cord herniation: value of MR phase-contrast imaging. AB - We report two patients with an idiopathic transdural spinal cord herniation at the thoracic level. Phase-contrast MR imaging was helpful in showing an absence of CSF flow ventral to the herniated cord and a normal CSF flow pattern dorsal to the cord, which excluded a compressive posterior arachnoid cyst. PMID- 10369370 TI - Another spurious description of hyperfixation of HMPAO. PMID- 10369371 TI - Three-dimensional power Doppler imaging of vertebrobasilar circulation in adults. PMID- 10369372 TI - Endovascular aneurysm treatment and the incidence of vasospasm. PMID- 10369374 TI - The importance of neuroradiologist authorship in nonspecialty journals. PMID- 10369373 TI - Direct angioplasty for acute occlusion of intracranial artery. PMID- 10369375 TI - The added value of neuroimaging for diagnosing dementia. PMID- 10369376 TI - Heterotopic neural tissue in the dural sinus: more frequent than usually assumed? PMID- 10369377 TI - Transbuccal space deep parotid biopsy. PMID- 10369378 TI - The consequences of traumatic brain injury on cerebral blood flow and autoregulation: a review. AB - In this decade, the brain argueably stands as one of the most exciting and challenging organs to study. Exciting in as far as that it remains an area of research vastly unknown and challenging due to the very nature of its anatomical design: the skull provides a formidable barrier and direct observations of intraparenchymal function in vivo are impractical. Moreover, traumatic brain injury (TBI) brings with it added complexities and nuances. The development of irreversible damage following TBI involves a plethora of biochemical events, including impairment of the cerebral vasculature, which render the brain at risk to secondary insults such as ischemia and intracranial hypertension. The present review will focus on alterations in the cerebrovasculature following TBI, and more specifically on changes in cerebral blood flow (CBF), mediators of CBF including local chemical mediators such as K+, pH and adenosine, endothelial mediators such as nitric oxide and neurogenic mediators such as catecholamines, as well as pressure autoregulation. It is emphasized that further research into these mechanisms may help attenuate the prevalence of secondary insults and therefore improve outcome following TBI. PMID- 10369379 TI - Enhanced nitric oxide synthesis reverses salt-induced alterations in blood flow and cGMP levels. AB - To understand the role of nitric oxide in salt-induced hypertension, we evaluated cardiovascular, hemodynamic and biochemical parameters in Dahl salt-sensitive rats fed low (0.3%) and high (8.0%) sodium diets. Two high salt groups received 1.25 and 2.5 g/L l-arginine in their drinking water. After three weeks of treatment, blood pressure was greater in the high salt groups. l-arginine did not modify salt-induced hypertension. Eicosapentaenoic acid (EPA) caused a smaller depressor response compared to normotensive rats. The increase in blood pressure was associated with decreases in aortic and renal blood flows. In renal artery, the reduction was counteracted by both l-arginine doses; whereas in the aorta, only the higher l-arginine one restored blood flow. The salt-induced reduction in aortic cyclic GMP level was only overcome by the higher l-arginine treatment. These data suggest that at the dose levels tested, nitric oxide reverses the reduction in cGMP and blood flow, but not the blood pressure changes associated with salt-induced hypertension. PMID- 10369381 TI - Interstrain differences in angiotensin I-converting enzyme mRNA and activity levels. Comparison between stroke-prone spontaneously hypertensive rats and Wistar-Kyoto rats. AB - Plasma angiotensin I-converting enzyme (ACE) levels are different between the stroke-prone spontaneously hypertensive rat (SHRSPHD) and the normotensive Wistar Kyoto (WKYHD) rat. This interstrain variability in plasma ACE levels is independent of blood pressure and is genetically linked to the ACE gene. The present study explored the hypothesis of an interstrain variability of tissue ACE activity and ACE gene expression levels. Tissue ACE levels were studied by enzymic activity measurement in the membrane fraction, and ACE mRNA levels were quantified by solution hybridization-ribonuclease protection assay. In lung, heart, kidney, and duodenum, membrane-bound ACE activity and ACE mRNA amount were significantly higher in WKYHD rats compared with SHRSPHD rats. No difference was observed in the testis where a specific isoform of the enzyme is produced. Our results suggest that in addition to determine differential plasma ACE levels between the WKYHD and SHRSPHD strains, the interstrain genetic variability also determines differential ACE mRNA and membrane-bound enzyme levels in somatic tissues. This likely reflects a difference in the ACE gene expression due to genetically determined regulatory mechanisms operative in all somatic tissues. PMID- 10369380 TI - Daily exercise enhances acetylcholine-induced dilation in mesenteric and hindlimb vasculature of hypertensive rats. AB - The effect of daily spontaneous running (DSR) on endothelial function was examined in spontaneously hypertensive rats (SHR). Following 8-11 weeks of DSR (n=15) or sedentary control (SED, n=15), rats were instrumented with arterial and venous catheters and mesenteric and iliac Doppler ultrasonic flow probes. Hemodynamic responses to vasodilator-mediated substances were determined under two experimental conditions; 1) bolus injection of indomethacin (10 mg/kg) and 4 bolus doses of acetylcholine (0.5-2.0 microg/kg); 2) bolus injection of N(omega) nitro-L-arginine (5 mg/kg) and 4 bolus doses of nitroglycerin (3-12 microg/kg). Hindlimb vascular conductance decreased more in response to indomethacin in DSR vs. SED rats (-18.3+/-2.8% vs. -10.4+/-2.5%). However, the mesenteric or hindlimb vascular conductance responses to N(omega)-nitro-L-arginine were not different between DSR and SED rats. DSR also enhanced mesenteric and hindlimb vascular conductance responses to acetylcholine. Results suggest that DSR enhances acetylcholine-induced vasodilation in SHR. PMID- 10369382 TI - Thapsigargin-insensitive calcium pools in vascular smooth muscle cells. AB - Since sarcoplasmic Ca2+-ATPase may play an important role for the regulation of cytosolic free calcium concentration ([Ca2+]i) and may be altered in primary hypertension, the effects of thapsigargin and bradykinin on intracellular calcium pools in cultured vascular smooth muscle cells (VSMC) from spontaneously hypertensive rats of the Munster strain (SHR) and normotensive Wistar-Kyoto (WKY) rats were investigated. VSMC were cultured on glass cover slips and [Ca2+]i was measured using the fluorescent dye fura2. To exclude transplasmamembrane calcium influx all experiments were performed in a calcium free medium. Thapsigargin, a selective inhibitor of the sarcoplasmic Ca2+-ATPase, and bradykinin, that is known to induce inositol trisphosphate release, dose dependently caused an increase of [Ca2+]i by emptying intracellular Ca2+ stores. The peak increase of [Ca2+]i after addition of saturation doses of thapsigargin (1 micromol/L) was not significantly different in the two strains (SHR: 69 +/- 11 nmol/L, n=24; WKY: 58 +/- 12 nmol/L, n=20; mean +/- SEM). When 10 micromol/L bradykinin was added after depletion of the thapsigargin-sensitive pools, still a release of [Ca2+]i could be observed. The bradykinin-induced [Ca2+]i increase was similar in the absence and presence of thapsigargin in VSMC from SHR (62 +/- 12 nmol/L, n=20; vs 52 +/- 18 nmol/L, n=22). In contrast, in the VSMC from WKY a significant reduction of the bradykinin induced [Ca2+]i-increase could be observed after the depletion of the thapsigargin sensitive calcium pools (70 +/- 8 nmol/L, n=21, vs. 33 +/- 7, n=20; p<0.002). It is concluded that bradykinin releases calcium from a pool that is not refilled by the common, thapsigargin-sensitive Ca2+-ATPase. In contrast to VSMC from normotensive WKY, in VSMC from spontaneously hypertensive rats thapsigargin and bradykinin sensitive pools may be regulated separately. PMID- 10369383 TI - Function of the isolated perfused kidneys from young or adult rats with post-DOCA salt hypertension. AB - The function of isolated perfused kidneys in post-DOCA phase of DOCA-salt hypertension was evaluated in rats subjected to DOCA-salt treatment either from youth or only in adulthood. Post-DOCA-salt hypertension was more severe in young than in adult animals. Kidneys isolated from young post-DOCA-salt hypertensive rats had higher renal vascular resistance when compared to adult ones. Perfusion of isolated kidneys over a wide range of perfusion pressure (110-170 mm Hg) has shown similar decrease of glomerular filtration, filtration fraction, diuresis and natriuresis in both age groups of DOCA-salt treated groups. Perfusion pressure-sodium excretion curves had reduced slope and were shifted to the right in both hypertensive groups, indicating huge glomerular damage. Nevertheless, the relative difference in glomerular filtration and sodium excretion between hypertensive rats and age-matched normotensive controls was always greater in younger animals. These results suggest that the more pronounced changes in glomerular hemodynamics and filtration could be involved in higher blood pressure response of young animals to DOCA-salt treatment. PMID- 10369384 TI - The effects of short-term nifedipine treatment on responsiveness of aortic rings of cadmium-hypertensive rats. AB - The effects of short-term antihypertensive treatment with nifedipine on blood pressure and vascular responsiveness were studied in cadmium-hypertensive and normotensive control rats. Cadmium administration caused a significant increase in mean arterial blood pressure. Endothelin-1, noradrenaline and angiotensin II produced concentration dependent contractions of aortic rings that attained a lower maximal contraction in cadmium-hypertensive rats. Responses of aortic rings to KCl did not show a significant difference between the groups. Nifedipine administered simultaneously with cadmium inhibited the induction of hypertension. Nifedipine treatment for 5 days significantly reduced the blood pressure in cadmium-hypertensive and normotensive rats. Neither inhibition of hypertension nor normalization of blood pressure in cadmium-hypertensive rats caused an alteration in contractile responses of aortic rings to vasoconstrictors which suggested that development of decreased vascular reactivity and of hypertension occurs simultaneously in cadmium-hypertensive rats but the role of decreased vascular reactivity in maintenance of hypertension is questionable in cadmium hypertension. PMID- 10369385 TI - Improvement in the capillarity of the left ventricular wall of stroke-prone spontaneously hypertensive rats following angiotensin II receptor blockade. AB - The effect of the angiotensin II type 1 receptor blockade, candesartan cilexetil (TCV116), on the capillarity of the innermost region of the left ventricular subendocardium were studied in stroke-prone spontaneously hypertensive rats (SHRSP). The rats were fed for 32 days on chow that contained TCV116, the average dose being 0.96 mg/kg/day. Compared with values from control SHRSP, the systolic blood pressure, left ventricular weight and cross-sectional area of cardiomyocytes decreased. A significant increase in the total capillary density was coupled with a decrease of capillary domain areas in all capillary portions. The proportion of venular capillary portions, which was low in control SHRSP, increased significantly, suggesting neoangiogenesis of capillaries. The results indicate that AT1-receptor blockade caused regression of the hypertrophied cardiomyocytes and improved capillarity of the left ventricular wall. PMID- 10369386 TI - Factors controlling epidermal growth factor (EGF) gene expression in the endometrium of the mare. AB - Previous studies showed a dramatic increase in EGF gene expression in the endometrial glands of pregnant mares around day 40 after ovulation. To investigate how the steroid hormones of pregnancy might regulate this expression, in situ hybridization was used to monitor the levels of EGF mRNA in endometrial biopsies obtained from seasonally anoestrous or ovariectomised mares given exogenous progesterone and oestrogen, alone or in combination, for up to 46 days. Biopsies were also taken from mares during the non-pregnant cycle, during normal pregnancies and pregnancies compromised by endometrial pathology (endometriosis) or because of incompatible extraspecific embryo transfers (donkey-in-horse pregnancies). Only a few samples showed weak EGF expression during the late luteal phase of the oestrous cycle. During normal pregnancy, the previously observed dramatic increase of expression after day 40 of gestation was confirmed. Although aged mares suffering from endometriosis and mares carrying an extraspecific donkey conceptus showed the same increase of EGF mRNA in normal glands, this was virtually absent from gland cross-sections compromised due to inflammatory or fibrotic changes. Administration of various doses and combinations of progesterone and oestrogen for < 35 days yielded negative or only weakly positive hybridization results, whereas progesterone alone for > or = 40 days upregulated EGF expression strongly irrespective of additional treatment with oestrogen. This is the first experimental evidence that EGF expression in the endometrium can be induced by progesterone alone. The requirement for prolonged progesterone priming is of considerable interest in the context of the unusually late stage of gestation at which placental attachment commences in equids. PMID- 10369387 TI - Sry-negative XX sex reversal in purebred dogs. AB - The gene responsible for testis induction in normal male mammals is the Y-linked Sry. However, there is increasing evidence that other genes may have testis determining properties. In XX sex reversal (XXSR), testis tissue develops in the absence of the Y chromosome. Previous polymerase chain reaction (PCR) assays indicated that autosomal recessive XXSR in the American cocker spaniel is Sry negative. In this study, genomic DNA from the breeding colony of American cocker spaniels and from privately owned purebred dogs were tested by PCR using canine primers for the Sry HMG box and by Southern blots probed with the complete canine Sry coding sequence. Sry was not detected by either method in genomic DNA of affected American cocker spaniels or in the majority (20/21) of affected privately owned purebred dogs. These results confirm that the autosomal recessive form of XXSR in the American cocker spaniel is Sry-negative. In combination with previous studies, this indicates that Sry-negative XXSR occurs in at least 15 dog breeds. The canine disorder may be genetically heterogeneous, potentially with a different mutation in each breed, and may provide several models for human Sry negative XXSR. A comparative approach to sex determination should be informative in defining the genetic and cellular mechanisms that are common to all mammals. PMID- 10369388 TI - Novel, testis-specific mRNA transcripts encoding N-terminally truncated choline acetyltransferase. AB - Previous studies reported the presence of choline acetyltransferase (ChAT) mRNA and protein in the mammalian testis. We have now found that none of the ChAT mRNAs produced in the testis is capable of encoding a full-length ChAT protein. Two ChAT cDNAs were isolated from an adult rat testis cDNA library encoding N terminally truncated ChAT proteins of 450 and 414 amino acids (aa), respectively, the former containing a novel N-terminal extension of 69 residues. Rapid Amplification of cDNA Ends (RACE) analysis revealed a complex pattern of 5' untranslated mRNA termini generated from the ChAT gene locus in the testis, all representing truncated versions of the ChAT enzyme. Two of these proteins were produced in transfected fibroblasts and found to lack ChAT activity. Neither did they show binding to the ChAT substrates, acetyl CoA and choline, in a competition assay. These results indicate that mammalian testis lacks a bona fide ChAT enzyme but expresses truncated ChAT proteins with a possible unique function to the testis. PMID- 10369389 TI - Expression and regulation of LRP-2/megalin in epithelial cells lining the efferent ducts and epididymis during postnatal development. AB - Low density lipoprotein receptor-related protein-2/megalin (LRP-2) is a receptor belonging to the low density lipoprotein receptor family that mediates endocytosis and lysosomal degradation of a variety of ligands including apolipoprotein J (Apo J)/clusterin/SGP-2. LRP-2 has been shown to be expressed regionally in the adult rat epididymis. In this study, we describe the pattern of expression of LRP-2 in the efferent ducts and epididymis during postnatal development of the rat and examine the role of testicular luminally derived substances on its expression. The expression of LRP-2 was analyzed immunocytochemically in tissues of normal animals ranging in age from postnatal day 7-90 and in 15-day-old efferent-duct-ligated animals sacrificed at later ages. In the efferent ducts, LRP-2 expression, appearing as a dense band on the apical surface of the nonciliated epithelial cells, was noted as early as day 7, well before the entry of sperm, Sertoli-cell-derived secretory products, and high levels of androgens. Efferent duct ligation studies further revealed that expression under this condition was comparable to controls at all later ages examined, suggesting that the factor regulating its expression was not a luminally derived testicular substance. In normal untreated animals, LRP-2 expression was not apparent at any of the ages examined in the proximal initial segment of the epididymis. By comparison, the distal initial segment, although having no LRP-2 expression from 7-15 days, showed expression in principal cells by day 21 which intensified at days 29 and 39. However, by day 49 and at later ages (56 and 90), LRP-2 immunoreactivity over principal cells became spotty or with weak or moderate reactivity in some cells and none in others. LRP-2 expression in the intermediate zone, proximal caput, corpus, and cauda regions also appeared in principal cells by day 21, intensified at days 29 and 39 and persisted as such at all later ages examined, correlating with high levels of androgens shown to occur by day 39. Although LRP-2 expression in the distal caput region was evident in principal cells at days 21 and 29, it became spotty with weak, moderate, or absent reactivity over principal cells at all later ages. These data suggest that LRP-2 expression is under the influence of both stimulatory and region-specific inhibitory factors. Analysis of 15-day-old efferent-duct-ligated animals at all later ages examined revealed that there was no change in LRP-2 expression along the entire epididymis, suggesting that both the stimulatory and inhibitory factors are not luminally derived testicular substances. The observed pattern of LRP-2 expression in all regions of the epididymis, except the distal caput region, was similar to that described for Apo J internalization by principal cells during postnatal development, showing a correlation between LRP-2 expression and its ligand, Apo J. In summary, LRP-2 expression in the epididymis undergoes region-specific changes during postnatal development and appears to be influenced by both stimulatory and inhibitory factors. PMID- 10369390 TI - Expression of mRNA encoding insulin-like growth factors I and II by uterine tissues and placenta during pregnancy in the rat. AB - The uterus and the placenta synthesize insulin-like growth factors (IGFs) and insulin-like binding proteins (IGFBPs). These growth factors are implicated in processes of proliferation and differentiation that occur in the uterus. To determine the patterns of expression of IGFs during rat pregnancy we used in situ hybridization with digoxigenin labeled probes on uterus from day 7 to day 16 of pregnancy. In early gestation days (7-8) both IGF mRNAs showed similar tissue distribution with relative abundance in the stroma and circular muscle layer. On days 11 and 12 expression for IGF-I mRNA was found in the mesometrial decidua and metrial gland and in the ectoplacental cone while clear expression of IGF-II mRNA could only be found in the latter. On days 13 and 14, expression for IGF-I mRNA could be detected in the mesometrial decidua and metrial gland but no expression was observed for IGF-II mRNA. A gradient of IGF-I mRNA expression could be observed in the placenta on day 16, with the trophoblastic cells of the basal zone expressing the signal with stronger intensity than in the labyrinthine zone. For IGF-II mRNA the highest expression was associated with the labyrinthine zone. Endovascular trophoblast was positive for both mRNAs. The spatial and temporal patterns of expression suggests a role for IGFs in the process of decidualization as well as in the establishment, growth and differentiation of the various trophoblast cells of the placenta. PMID- 10369391 TI - Cellular and subcellular localization of stathmin during oocyte and preimplantation embryo development. AB - Stathmin is a 19 kDa cytosolic phosphoprotein, proposed to act as a relay integrating diverse intracellular signaling pathways involved in regulation of cell proliferation, differentiation, and function. To gain further information about its significance during early development, we analyzed stathmin expression and subcellular localization in mouse oocytes and preimplantation embryos. RT-PCR analysis revealed a low expression of stathmin mRNA in unfertilized oocytes and a higher expression at the blastocyst stage. A fine cytoplasmic punctuate fluorescent immunoreactive stathmin pattern was detected in the oocyte, while it evolved toward an increasingly speckled pattern in the two-cell and later four- to eight-cell embryo, with even larger speckles at the morula stage. In blastocysts, stathmin immunoreactivity was fine and intense in inner cell mass cells, whereas it was low and variable in trophectodermal cells. Electron microscopic analysis allowed visualization with more detail of two types of stathmin immunolocalization: small clusters in the cytoplasm of oocytes and blastocyst cells, together with loosely arranged clusters around the outer membrane of cytoplasmic vesicles, corresponding to the immunofluorescent speckles in embryos until the morula stage. In conclusion, it appears from our results that maternal stathmin is accumulated in the oocyte and is relocalized within the oocyte and early preimplantation embryonic cell cytoplasm to interact with specific cytoplasmic membrane formations. Probably newly synthesized, embryonic stathmin is expressed in the blastocyst, where it is localized more uniformly in the cytoplasm mostly of inner cell mass (ICM) cells. These expression and localization patterns are probably related to the particular roles of stathmin at the successive steps of oocyte maturation and early embryonic development. They further support the proposed physiologic importance of stathmin in essential biologic regulation. PMID- 10369392 TI - Timing of the first cleavage post-insemination affects cryosurvival of in vitro produced bovine blastocysts. AB - The time of the first cleavage of bovine zygotes during in vitro culture can affect the rate of development and cell number of the blastocysts. The aim of this work was to study the effect of the timing of first cleavage on the cryosurvival of the resulting blastocysts. Following standard IVM and IVF, zygotes were cultured in modified synthetic oviduct fluid (SOF), with 10% fetal calf serum (FCS) added 48 hr post insemination, in a humidified atmosphere of 5% CO2, 5% O2 and 90% N2. Embryos which cleaved by 24, 27, 30, 33, or 36 hr after insemination (IVF) were harvested and further cultured to the blastocyst stage (day 7 or day 8 post IVF). All developing blastocysts on days 7 and 8 were classified into three groups and were cryopreserved by vitrification. Group A consisted of blastocysts (<150 microm, small blastocysts); group B consisted of expanded or hatching blastocysts (>150 microm, large blastocysts); and group C consisted of morphologically poor quality blastocysts. The vitrification solution consisted of 6.5 M glycerol and 6% bovine serum albumin in PBS (VS3a). Thawed embryos were cultured further and survival was defined as the re-expansion and maintenance of the blastocoel over 24, 48, and 72 hr, respectively. Overall survival and hatching at 72 hr post-thawing was higher in blastocysts formed by day 7 than those formed by day 8 (60% vs. 40% survival; 63% vs. 45% hatching). Large blastocysts from day-7 and day-8 groups survived significantly better than small or poor quality blastocysts (76% vs. 63% and 31%; 72% vs. 30% and 26%, respectively; P < 0.05). Day-7 blastocysts from the 27- and 30-hr cleavage groups survived significantly better than those from the 36-hr group (63% and 66% vs. 25%, P < 0.05). Day-8 blastocysts from later cleaved (30 hr) zygotes had a higher survival than the 27-hr cleavage groups (52% vs. 26%, P < 0.05). These results indicate that the day of blastocyst appearance, developmental stage, and timing of the first cleavage post-insemination can influence the cryosurvival of bovine blastocysts following vitrification. PMID- 10369393 TI - Ultrastructure of bovine embryos developed from in vitro-matured and -fertilized oocytes: comparative morphological evaluation of embryos cultured either in serum free medium or in serum-supplemented medium. AB - The ultrastructure of bovine embryos developed from in vitro-matured and fertilized oocytes, cocultured with bovine cumulus/granulosa cells either in a serum-free medium (IVMD101) or in a serum-containing medium (TCM199+CS) was compared. Embryos up to the eight-cell stage had many cellular organelles and cytoplasmic components that were randomly distributed in the cytoplasm. Mitochondria were spherical or ovoid and had only a few peripheral cristae. There were no obvious differences in the ultrastructure between embryos developed in IVMD101 and TCM199+CS up to the eight-cell stage. However, conspicuous differences in the ultrastructural features between the embryos cultured in IVMD101 and TCM199+CS were observed at the morula and blastocyst stages. At the morula stage, embryos cultured in IVMD101 had cells containing elongated mitochondria, well-developed Golgi apparatus, lipid droplets, and large vesicles resembling lysosomes. The lysosome-like vesicles were partially filled with electron-dense materials and were frequently fused with lipid droplets. The blastomeres of morulae cultured in TCM199+CS contained numerous large lipid droplets and fewer lysosome-like vesicles than those cultured in IVMD101. In blastocysts cultured in IVMD101, lysosome-like vesicles were frequently observed in the trophoblast cells and lipid droplets were present in the cytoplasm of trophoblast and inner cell mass (ICM)-cells, but they were not abundant. On the other hand, the blastocysts developed in TCM199+CS contained fewer lysosome-like vesicles and large numbers of lipid droplets. This accumulation of lipid droplets was higher in the trophoblast cells than in the ICM-cells. This study showed major differences in the ultrastructural features between the morulae and blastocysts from serum-free and serum-supplemented cultures, suggesting that the ultrastructural differences may reflect physiological characteristics of embryos. PMID- 10369394 TI - Thermotolerance of IVM-derived bovine oocytes and embryos after short-term heat shock. AB - A series of experiments were designed to study the effect of elevated temperatures on developmental competence of bovine oocytes and embryos produced in vitro. In experiment 1, the effect of heat shock (HS) by a mild elevated temperature (40.5 degrees C) for 0, 30, or 60 min on the viability of in vitro matured (IVM) oocytes was tested following in vitro fertilization (IVF) and culture. No significant difference was observed between the control (39 degrees C) and the heat-treated groups in cleavage, blastocyst formation, or hatching (P > 0.05). In experiment 2, when the HS temperature was increased to 41.5 degrees C, neither the cleavage rate nor blastocyst development was affected by treatment. However, the rate of blastocyst hatching appeared lower in the HS groups (13% in control group vs. 3.9% and 5.6% in 30 min and 60 min, respectively; P < 0.05). When IVM oocytes were treated at 43 degrees C prior to IVF (experiment 3), no difference was detected in blastocyst and expanded blastocyst development following heat treatment for 0, 15, or 30 min, but heat treatment of oocytes for 45 or 60 min significantly reduced blastocyst and expanded blastocyst formation (P < 0.05). In experiment 4, the thermotolerance of day 3 and day 4 bovine IVF embryos were compared. When embryos were pre-treated with a mild elevated temperature (40.5 degrees C) for 1 hr, and then with a higher temperature (43 degrees C) for 1 hr, no improvement in thermotolerance of the embryos was observed as compared to those treated at 43 degrees C alone. However, a higher thermotolerance was observed in day 4 than day 3 embryos. In conclusion, treatment at 43 degrees C, but not 40.5 degrees C or 41.5 degrees C significantly reduced oocyte developmental competence. An increase in thermotolerance was observed from day 3 to day 4 of in vitro embryonic development, which corresponds to the maternal to zygotic transition of gene expression in bovine embryos. PMID- 10369395 TI - Rise of intracellular Ca2+ level causes the decrease of cyclin B1 and Mos in the newt eggs at fertilization. AB - Unfertilized eggs of the newt, Cynops pyrrhogaster, are arrested at the second meiotic metaphase, with activity of the M-phase promoting factor (MPF) maintained at a high level. After fertilization, the eggs resume the cell cycle, and emit the second polar body. When the change in [Ca2+]i in the fertilized eggs was monitored by aequorin, an early increase in [Ca2+]i was observed 5-10 min after insemination and continued for about 30 sec. A late increase in [Ca2+]i then occurred 10-15 min after fertilization and continued for 30-40 min. The injection of 1,2-Bis (2 aminophenoxy) ethane-N,N,N',N',-tetraacetic acid (BAPTA) into unfertilized eggs inhibited reinitiation of the cell cycle after fertilization. Western blot analysis with antibodies against cyclin B1 or Mos indicated that both cyclin B1 and Mos were present in unfertilized eggs, but both disappeared within 30 min after fertilization. Treatment with Ca2+-ionophore decreased both cyclin B1 and Mos. Chymotryptic activity in Cynops egg extracts was not significantly increased after fertilization or activation by treatment with the Ca2+-ionophore. No change in [Ca2+]i was observed following treatment with cycloheximide, but the amount of both cyclin B1 and Mos rapidly decreased. These results indicate that resumption of meiosis in Cynops eggs is induced by an increase in [Ca2+]i at fertilization, which causes degradation of both cyclin B1 and Mos by inhibition of de novo synthesis of those proteins. PMID- 10369397 TI - Preservation of endangered species and populations: a role for genome banking, somatic cell cloning, and androgenesis? PMID- 10369396 TI - PH-20 but not acrosin is involved in sperm penetration of the macaque zona pellucida. AB - In this study, we investigated the functions of PH-20 and acrosin during the interaction of macaque sperm with the zona pellucida. Both of these sperm enzymes have been reported to be present on the inner acrosomal membrane of acrosome reacted sperm, and have been suggested to play a role during secondary sperm-zona binding in other species. Anti-macaque PH-20 IgG, anti-pig acrosin IgG and soybean trypsin inhibitor (SBTI) were used as probes for immunolocalization of the two proteins at the ultrastructural level, and as reagents for blocking sperm penetration of the macaque zona pellucida in vitro. As a control, we performed similar studies with antibodies to CD-46, which is also located on the inner acrosomal membrane, but has no known function in sperm-zona pellucida interaction. After labeling with anti-acrosin IgG, gold label was not present on the sperm surface before the acrosome reaction, but was detected over the entire head of sperm that were induced to acrosome react with calcium ionophore A23187. In contrast, when sperm were induced to acrosome react by binding to intact zona pellucida, acrosin was present in the acrosomal shroud but not on the inner acrosomal membrane. Similar results were obtained when SBTI was used as a probe for enzyme localization. PH-20 and CD-46 were demonstrated on the inner acrosomal membrane of sperm induced to acrosome react by ionophore treatment and by zona binding. Neither anti-acrosin IgG nor anti-CD-46 IgG affected sperm penetration of the zona at concentrations up to 300 microg/ml, but zona penetration was blocked completely when anti-PH-20 IgG (100 microg/ml) was present during sperm oocyte interaction. Ultrastructural observations of oocytes incubated with anti PH-20 IgG showed that acrosomal shrouds were present on the zona surface but no sperm had begun to penetrate into the zona substance. We conclude that anti-PH-20 IgG prevented sperm penetration of the macaque zona pellucida by interference with secondary sperm-zona binding, rather than primary sperm-zona binding or the zona-induced acrosome reaction. Acrosin was not detected on the inner acrosomal membrane of sperm that are induced to acrosome react after zona binding, and acrosin does not appear to be critical for sperm penetration of the macaque zona pellucida. PMID- 10369398 TI - Overview of nimesulide. PMID- 10369399 TI - Relationship of nimesulide safety to its pharmacokinetics: assessment of adverse reactions. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) are among the most widely used drugs and their use is frequently associated with severe or even serious adverse events, which increase morbidity and mortality. The increase of toxic effects, primarily of the digestive system, due to treatment with NSAIDs, underlines a need for safer NSAIDs. Nimesulide (4-nitro-2-phenoxymethanesulphonanilide) is a chemically unique anti-inflammatory agent in that it has a higher pKa (6.5) than conventional acidic NSAIDs and it is one of the newer class of NSAIDs that are selective for cyclooxygenase-2. Nimesulide also has additional activities, among them effects on production/action of oxy-radicals and other components of neutrophil activation that may contribute to the spectrum of its anti inflammatory activity as well as potentially reducing the likelihood of gastrointestinal ulcerogenicity. An analysis was performed of the safety data of nimesulide collected in clinical studies and from those reported spontaneously worldwide in the post-marketing phase. The results show that nimesulide is associated with a relatively low occurrence of adverse drug reactions especially in the gastrointestinal tract while those in the liver are within or below the general incidence with other NSAIDs. PMID- 10369400 TI - Effect of nimesulide on glucocorticoid receptor activity in human synovial fibroblasts. AB - Fibroblasts from human synovial membranes were cultured with nimesulide, naproxen or dexamethasone. Nimesulide, but not naproxen, showed effects on the glucocorticoid system that may contribute importantly to its anti-inflammatory activity. Nimesulide at therapeutically relevant concentrations induced the intracellular phosphorylation and activation of glucocorticoid receptors, and activated their binding to the target genes. Naproxen or dexamethasone markedly reduced the number of glucocorticoid receptor binding sites, in contrast to nimesulide, which had no significant effect. PMID- 10369401 TI - Molecular model of the interaction between nimesulide and human cyclooxygenase-2. AB - The cyclooxygenase-2 (COX-2) isoenzyme is a key target for COX-2-selective non steroidal anti-inflammatory drugs (NSAIDs). An important difference in binding of nimesulide compared with non-selective NSAIDs appears to involve the amino acid at position 523 of the enzyme. Replacement of valine with isoleucine at this position provides access to a binding site that is larger in COX-2 than in COX-1. Nimesulide appears to exploit this enlarged binding site for establishing a number of favourable contacts with the enzyme that lead to selective inhibition of COX-2. We made these conclusions from a three-dimensional molecular model of the active site of human COX-2, constructed using the X-ray coordinates of COX-1 from sheep seminal vesicles and COX-2 from mouse fibroblasts as templates, with the aid of sequence alignment methods and molecular modelling techniques. The resulting model was refined, and the active site was probed for regions of steric and electrostatic complementarity for ligand binding. Docking studies were then undertaken with many different nimesulide conformers, a family of which could establish very favourable interactions with the NSAID binding site of human COX-2 by exploiting the extra space made available by the isoleucine/valine replacement. The stability of the resulting complexes was studied by simulating molecular dynamics. PMID- 10369402 TI - The in vivo assessment of nimesulide cyclooxygenase-2 selectivity. AB - In man, nimesulide selectively inhibits cyclooxygenase-2 (COX-2) with little effect on haemostatic function or gastric prostaglandin formation. It causes significantly less gastrointestinal injury than naproxen, but has anti inflammatory efficacy similar to that of naproxen and other currently available non-steroidal anti-inflammatory drugs. Naproxen suppressed arachidonic-acid mediated platelet aggregation, reduced serum thromboxane B2 levels by 98% throughout the treatment period and reduced gastric mucosal prostaglandins (PGE2 and 6-keto-PGF1alpha) production by an average of 80%. This contrasts with nimesulide: platelet aggregation was not significantly affected, thromboxane B2 levels were only 29% lower and the gastric mucosal prostaglandins were inhibited in the order of approximately 20%. During the treatment period, both nimesulide and naproxen significantly inhibited COX-2-dependent PGE2 synthesis in the whole blood; however, naproxen had a lesser effect than nimesulide. PMID- 10369403 TI - Gastrointestinal toxicity of non-steroidal anti-inflammatory drugs: the effect of nimesulide compared with naproxen on the human gastrointestinal tract. AB - This overview includes theories and evaluation of non-steroidal anti-inflammatory drug (NSAID)-induced gastrointestinal toxicity. Factors in damage include microvascular aspects, neutrophil recruitment, mucosal prostaglandins, gastrointestinal secretions and bacteria. We have proposed an extensive simplified framework that includes an important local initiating effect which may involve NSAID accumulation, interaction with surface phospholipids, events that alter cellular ATP, and local/systemic effects of cyclooxygenase (COX) inhibition. COX-2-selective drugs are desirable not only because they spare COX-1 and so avoid gastrointestinal toxicity, but also because COX-2-selective agents are only weakly acidic and therefore avoid substantial accumulation in the gastric mucosa. Short-term endoscopy studies of NSAIDs are important initially to evaluate human gastroduodenal tolerability. They show that injury increases with the amount of NSAIDs even though the lowest therapeutic doses inhibit gastric COX almost completely, and that the more-acidic NSAIDs tend to cause greater gastric damage. Long-term endoscopy studies involve NSAID ingestion for at least 3 months. A main question is the extent to which the ulcers seen cause symptoms, substantial bleeding and/or perforation. Measurement of serious outcomes is thought by many to be the best assessment of gastrointestinal safety, but studies find marked variations even with the same drug. Damage to the small intestine by NSAIDs is even more frequent than to the upper gastrointestinal tract, but is difficult to evaluate. Conventional acidic NSAIDs increase the permeability of human small intestine, probably by a non-prostaglandin mechanism, but nimesulide does not do so, possibly because of its very weak acidity. PMID- 10369405 TI - Economic comparison of nimesulide and diclofenac, and the incidence of adverse events in the treatment of rheumatic disease in Greece. AB - In Greece a 15-day treatment of rheumatic disease with diclofenac costs 56% more than treatment with nimesulide. This is due to the lower incidence of gastrointestinal adverse events with nimesulide, and the absence of serious gastrointestinal complications leading to hospitalization, which more than offset the higher acquisition cost of nimesulide. The average saving by using nimesulide instead of diclofenac is about US$21 per patient for a 15-day treatment period. PMID- 10369404 TI - Comparative efficacy and safety of nimesulide and diclofenac in patients with acute shoulder, and a meta-analysis of controlled studies with nimesulide. AB - Adverse events, particularly gastrointestinal, partially offset the therapeutic value of NSAIDs. The abilities of nimesulide to inhibit COX-2 preferentially and to exert other novel anti-inflammatory actions are consistent with good efficacy and safety. This is borne out by a double-blind multicentre comparison of nimesulide and diclofenac in 122 patients with acute shoulder, and by a meta analysis of various nimesulide trials. At the end of the 14 day double-blind study, nimesulide was at least as effective as diclofenac (investigator ratings: good/very good in 79.0% of patients given nimesulide, and 78.0% with diclofenac; patient ratings: good/very good in 82.3 and 78.0% respectively). Four patients (6.5%) dropped out in the nimesulide group (two early recovery, one lack of effect, one adverse event), compared with 13 (21.7%) in the diclofenac group, due mainly to adverse events (P=0.003). Global tolerability was judged by the investigators to be good/very good in 96.8% of the nimesulide group compared with 72.9% of those given diclofenac. Judgements by the patients were 96.8 and 78.0% respectively. Both differences are highly significant statistically. The meta analysis demonstrates that nimesulide given for 2 weeks is far more efficacious than placebo in treating osteoarthritis, and is at least comparable to other NSAIDs The benefit-risk ratio for nimesulide was better in all individual studies since 100 mg nimesulide twice daily was about equal to placebo in safety and tolerability, especially regarding gastrointestinal adverse events. PMID- 10369407 TI - Expression of sex hormone-binding globulin exon 7 splicing variant mRNA in secondary spreading lesions of gynecological cancers. AB - This study was designed to determine the clinical implications of intracellular expression of sex hormone-binding globulin (SHBG) wild-type and exon 7 splicing variant mRNAs in secondary spreading lesions of gynecologic cancers using the reverse transcription-polymerase chain reaction-Southern blot and DNA sequencing analyses. Compared with primary cancers, a relative increase in SHBG variant mRNA to wild-type mRNA expression was observed (4/10 cases of uterine endometrial cancers, 5/10 cases of uterine cervical cancers, 6/10 cases of ovarian cancers) or the expression of SHBG wild-type and variant mRNAs could not be detected (5/10 cases of uterine endometrial cancers, 3/10 cases of uterine cervical cancers, 4/10 cases of ovarian cancers). On the other hand, alteration to a relative increase in SHBG wild-type mRNA expression in the metastatic lesions occurred in only 3 cases (1/10 cases of uterine endometrial cancers and 2/10 cases of uterine cervical cancers) analyzed. These results suggest that the transcription of SHBG mRNA and the regulation of its splicing might be altered with metastatic potential, and this status might be involved in a change in steroidal dependency of metastatic lesions. PMID- 10369406 TI - Quantity of partial agonist activity for antiglucocorticoids complexed with mutant glucocorticoid receptors is constant in two different transactivation assays but not predictable from steroid structure. AB - An unsolved question in steroid hormone action is why the amount of agonist activity displayed by antisteroids is not constant but varies with the assay conditions. Receptor mutations have provided insight into hormone action, presumably due to changes in the tertiary structure of the receptor that alter its interaction surfaces with the transcriptional machinery or/and co-factors. We have now employed two mechanistically different induction assays to determine whether disparate transactivation processes are similarly altered by receptor mutations. The two activation assays studied were (i) the standard induction of GREtkLUC in transiently transfected CV-1 cells and (ii) a novel modulation of endogenous receptor activity by transiently transfected receptors in HeLa cells. Five different mutations in the ligand binding and DNA binding domains of the rat glucocorticoid receptor (CS1, CS1/CD, 451/9, C656G, and R732Q) and seven steroids of varied structures (five antagonists and two agonists) were selected for use. The results in both induction assays were the same. However, no generalizations regarding steroid structure and activity emerged. Neither of two potent glucocorticoids were active with GR-CS1, or GR-CS1/CD, while RU 486 was the only antisteroid with appreciable agonist activity. With the GR-451/9 mutant, three antagonists afforded partial agonist activity. We confirmed that the C656G mutant is both "super-sensitive" and "super-selective" for transactivation. In contrast, the R732Q mutation caused significant decreases in activity with both antagonists and subsaturating concentrations of agonists. This inability to generalize about the behavior of any class of steroids with mutant receptors may reflect an induced fit for each receptor steroid complex. Nevertheless, the activity of a given steroid appeared to be constant in two different transactivation assays for a given mutant receptor. Thus, disparate transactivation processes may utilize identical receptor surfaces, even in the expression of partial agonist activity for specific antiglucocorticoids. PMID- 10369408 TI - Sex hormone binding globulin expression and colocalization with estrogen receptor in the human Fallopian tube. AB - The detection of sex hormone binding globulin (SHBG) or SHBG mRNA in several sex steroid target tissues, has raised the possibility that SHBG modulates the action of sex steroids outside the vascular compartment. The presence of SHBG mRNA was investigated by RT-PCR in the poly (A+) RNA fraction of the human Fallopian tube. Human and rat liver were used as positive and negative control tissues, respectively. The electrophoretic analysis of the amplified PCR products showed bands at 219 bp, corresponding to the expected size of the SHBG cDNA, in the Fallopian tube and human liver but not in rat liver, indicating that SHBG might be synthesized by the Fallopian tube. The cellular localization of SHBG and of estrogen receptor (ER) was examined by immunohistochemistry in consecutive sections of Fallopian tube tissues for individual staining or double immunostaining in the same section. Specific immunostaining of SHBG was present in the epithelial, vascular and muscle cells of the ampullary and isthmic region. In epithelial cells, immunoreactive SHBG was present in the apical end with the highest concentration close to the luminal membrane. The ER was localized in the nuclei of epithelial, stromal and muscle cells of the ampulla and isthmus. Double immunostaining showed that SHBG and ER are colocalized principally in epithelial cells of the ampulla and in muscle cells of the isthmus. In conclusion, the detection of SHBG and SHBG mRNA and the localization of SHBG in estrogen target cells was shown. These findings support the hypothesis that SHBG might regulate sex steroid action at the tissue level. PMID- 10369409 TI - Heterologous expression of wild type and deglycosylated human sex steroid-binding protein (SBP or SHBG) in the yeast, Pichia pastoris. Characterization of the recombinant proteins. AB - Wild type, partially and fully-deglycosylated human sex steroid-binding protein (SBP or SHBG) cDNAs lacking the native cucaryotic signal sequence were cloned into a yeast expression vector containing the Saccharomyces cerevisiae alpha factor for extracellular secretion. Following transformation into Pichia pastoris, the wild type and all constructed mutants were successfully expressed. The levels were lower for the deglycosylated mutants indicating that oligosaccharide side chains may play a role in SBP secretion. Under fermentation conditions, the wild type protein was expressed at a level of 4 mg/l while the fully-deglycosylated mutant T7A/N351Q/N367Q was expressed at about 1.5 mg/l. The latter was purified from several fermentation runs and was found to be completely deglycosylated, electrophoretically homogeneous and fully active. The aminoterminus was found to have the sequence NH2QSAHDPPAV- indicating that cleavage of the alpha-factor occurred at the A(+7)-Q(+8) peptide bond. The molecular mass of the subunit was determined to be 39,717.8 Da, which is in complete agreement with the amino acid sequence of the T7A/N351Q/N367/Q mutant. The equilibrium constants for the dissociation of 5alpha-dihydrotestosterone and steroid binding specificity were found to be identical to that of the human plasma protein indicating that the missing N-terminal segment NH2-LRPVLPT and the removal of oligosaccharide side chains do not affect the stability and active conformation of the protein. In conclusion, the data presented reveal that the SBP mutant T7A/N351Q/N367/Q is the protein of choice for solving the three dimensional structure. PMID- 10369410 TI - The regulation of oestrone sulphate formation in breast cancer cells. AB - The formation of oestrone sulphate has been examined in MCF-7 (oestrogen receptor positive, ER+) and MDA-MB-231 (ER negative, ER-) breast cancer cells. Using intact cell monolayers and a physiological substrate concentration, progesterone (1 microM) and dexamethasone (1 microM) both increased oestrone sulphate formation in MCF-7 cells. In MDA-MB-231 cells, dexamethasone, but not progesterone, increased conjugate formation. A number of growth factors, cytokines and human serum albumin (HSA), which have previously been found to regulate oestrogen synthesis, were also examined for their ability to regulate oestrone sulphate formation. In MCF-7 cells epidermal growth factor, acidic and basic fibroblast growth factors, insulin-like growth factor-type I and insulin all stimulated oestrone sulphate formation. The cytokines, tumour necrosis factor alpha (TNFalpha) and interleukin-1beta also increased conjugate formation in the ER+ cells, as did HSA. In contrast, in MDA-MB-231 cells TNFalpha was without effect and HSA inhibited oestrone sulphate formation. The ability to modulate oestrone sulphate formation in ER+ cells may be an important mechanism to limit the availability of oestrogen to interact with the ER. PMID- 10369411 TI - Regulation of estrogen activity by sulfation in human Ishikawa endometrial adenocarcinoma cells. AB - Sulfation is an important conjugation reaction in the metabolism of steroids. Steroids sulfates do not interact with the appropriate hormone receptors; additionally, the presence of the charged sulfate moiety increases the aqueous solubility and excretion of most steroids. Estrogen sulfotransferase (EST) is the major form of human cytosolic ST involved in the conjugation of estrogens. EST is important in the inactivation of beta-estradiol (E2) during the luteal phase of the menstrual cycle. EST has a significantly higher affinity for the sulfation of E2 and 17alpha-ethinylestradiol (EE2) than for other potent estrogens such as diethylstilbestrol (DES) and equine estrogens. The ability of EST to sulfate these estrogenic compounds at physiologic concentrations is important in regulating their activation of the ER in estrogen responsive cells. Human Ishikawa endometrial adenocarcinoma (ISH) cells possess an estrogen receptor (ER) regulated alkaline phosphatase (AlkPhos) which is used to assay ER activation. To study the effects of EST activity on the ER activation of different estrogenic compounds, ISH cells were stably transformed with an EST expression vector. Dose response curves for the induction of AlkPhos activity by the different estrogenic compounds were generated with EST/ISH and control pcDNA/ISH cells. EST/ISH cells were 200-fold less sensitive to E2 and EE2 than were control cells. No differences were observed in the dose response curves for DES between EST/ISH and pcDNA/ISH cells. EST/ISH cells were approximately 3-10-fold less sensitive to the equine estrogens equilin and 17-equilin as compared to control cells. The ability of EST to decrease the ER activation of an estrogen correlates with the sulfation of these compounds at nanomolar concentrations by EST/ISH and pcDNA/ISH ISH cells. These results indicate that EST is capable of efficiently inactivating E2 and EE2 but is significantly less effective in inhibiting the ER binding of other potent estrogenic compounds. PMID- 10369412 TI - Interaction between estrogen receptor and Pit-1 protein is influenced by estrogen in pituitary cells. AB - The estrogen responsiveness of the rat prolactin gene expression requires the presence of both the estrogen receptor (ER) and the tissue-specific transcription factor, Pit-1 protein. We performed protein interaction assays using anti-rat Pit 1 antiserum (a-rPit-1) to investigate the physical interactions which occur between ER and Pit-1 proteins following estrogen treatment. After fusing maltose binding protein (MBP) and Pit-1 protein, we used the resulting MBP Pit-1 fusion protein to prepare a-rPit-1. Our results show that the estrogen receptor readily co-precipitated with the Pit-1 protein drawn from the lysates of two prolactin expressing pituitary cell lines GH3 and PR1. The rate of precipitation appears to be both estrogen- and time-dependent. Cellular levels of estrogen receptors and Pit-1 proteins did not show significant changes during the time of estrogen treatment. We therefore suggest that an estrogen-dependent physical interaction between ER and Pit-1 protein exists in vivo, and that this interaction may play an important role in the regulation of prolactin gene expression. PMID- 10369413 TI - Role of proteoglycans on testosterone synthesis by purified Leydig cells from immature and mature rats. AB - In order to characterize an involvement of proteoglycans (PG) in the regulation of Leydig cell function, we have examined the effects of para-nitrophenyl-beta-D xyloside (PNPX), a specific inhibitor of PG synthesis and para-nitrophenyl-beta-D galactoside (PNPG), an inefficient structural analogue, on testosterone production by purified Leydig cells from immature and mature rats, in the presence or not of various concentrations of hCG during 24 h. Whatever the age, the addition of PNPX induces a decrease of [35S] and [3H] incorporations into cell layer associated-PG; these latter being less numerous (-50 and -25%, respectively in immature and mature rat), and less sulfated (-40%) when compared to control Leydig cells. In immature Leydig cells, the inhibition of PG synthesis decreases both the basal and weakly stimulable-hCG or -(Bu)2cAMP or -LH testosterone synthesis. In mature Leydig cells, the PG inhibition has no effect on testosterone production both in the absence of hCG and in the presence of weak amounts of hCG but increases it in the presence of subsaturating hCG concentrations. Whatever the age, the inhibition of PG synthesis is ineffective in the presence of saturating amounts of either hCG or (Bu)2cAMP. These effects are maintained in the presence of MIX, PMA, but are not observed in the presence of 22R-hydroxycholesterol. Therefore, our results suggest that in rat Leydig cells, the inhibition of PG synthesis affects the signal transduction at a step distal to cyclic AMP and more precisely, the cholesterol supply to the mitochondria by acting on its cellular distribution (free and esterified cholesterol). PMID- 10369414 TI - EcR interacts with corepressors and harbours an autonomous silencing domain functional in both Drosophila and vertebrate cells. AB - The ecdysone receptor (EcR) is a member of the large family of nuclear hormone receptors, which are ligand regulated transcription factors. In general, ligand converts these receptors into a transcriptional activator. Some vertebrate nuclear hormone receptors, such as the thyroid hormone and retinoic acid receptors, silence gene expression in the absence of ligand. EcR is involved in fly metamorphosis and is used in vertebrates as an inducible system for expression of transgenes. Here, we show that a Drosophila receptor, the EcR, harbours an autonomous silencing function in its carboxy-terminus. Interestingly, EcR mediates also silencing in vertebrate cells. In concordance with this EcR interacts with the corepressors SMRT and N-CoR, while addition of ligand reduces this interaction. Conversely, the v-erbA oncogene product, a thyroid hormone receptor derivative, mediates silencing in Drosophila cells. Thus, our data suggest the involvement of an evolutionarily conserved mechanism by which nuclear hormone receptors mediate gene silencing in multicellular organisms. PMID- 10369415 TI - Membrane anchoring of calnexin facilitates its interaction with its targets. AB - Calnexin, a chaperone that resides in the endoplasmic reticulum, participates in the quality control function of this compartment. Many glycoproteins in the process of folding associate transiently with this chaperone via interactions involving the recognition of their mono-glucosylated glycans. Some misfolded proteins which are retained in the endoplasmic reticulum exhibit prolonged association with calnexin. We have examined whether the transmembrane and cytoplasmic domains of calnexin influence the association of this chaperone with its targets. Interactions of wild type and truncated calnexin with a glycoprotein that is retained in the endoplasmic reticulum (the lymphocyte tyrosine kinase, Ltk), with membrane IgM heavy chains, and with the MHC class I heavy chain protein were investigated. A soluble calnexin molecule lacking the transmembrane domain and cytoplasmic tail does not associate with any of these proteins. When a heterologous transmembrane domain is fused to the lumenal portion of calnexin, this membrane-bound protein can bind Ltk, IgM heavy chains, and MHC class I heavy chain proteins. These results suggest that calnexin must be membrane-anchored in order to recognize its substrates. PMID- 10369416 TI - A model of multivalent ligand-receptor equilibria which explains the effect of multivalent binding inhibitors. AB - Quantitative relationships based on multivalent ligand receptor binding equilibria are developed which can describe the enhanced binding observed when polyvalent ligands are used as inhibitors of cell/cell interactions. This theory is able to explain the many orders of magnitude difference reported between the apparent dissociation constants determined for the interaction of Entamoeba histolytica membrane receptors with mono and multivalent N-Acetylgalactosaminide inhibitors. Given two experimentally accessible constants the theory provides a framework for the quantitative evaluation of custom designed polyvalent inhibitors of lectin and antibody mediated interactions. PMID- 10369417 TI - Temporal expression of a V(H) promoter-Cmu transgene linked to the IgH HS1,2 enhancer. AB - Immunoglobulin heavy chain (IgH) gene expression is guided by cis-regulatory elements which direct correct temporal and spatial expression in B lineage cells. One of these cis-acting elements is the IgH HS1,2 enhancer and previous studies in transgenic mice have revealed a temporally restricted activity of an HS1,2 enhancer-linked human beta-globin reporter gene in B lineage cells. To assess whether this enhancer can impose strict temporal regulation onto a V(H)-promoter Cmu reporter gene, transgenic mice were generated. These mice expressed high serum levels of protein from the transgene. Moreover, high levels of transgene expression were observed in spleen and thymus, while lower expression was found in heart and kidney and no expression was detected in liver and brain. Interestingly, transgene expression was confined to large, activated B cells and peritoneal B cells but not observed in small, resting splenic B cells or activated T cells. However, upon mitogenic stimulation of resting B cells with LPS, high levels of transgene expression was induced. Our data demonstrate that the HS1,2 enhancer can interact with a natural V(H) promoter in a strict temporal fashion and when provided with an appropriate activation signal, this V(H) promoter/enhancer construct can induce transgene expression in resting B, but not T lineage cells. Our data are compatible with a model whereby the regulation of IgH gene expression may be subject to regulation by distinct subsets of cis regulatory elements acting at different stages of B lymphocyte development. Thus, Ig gene expression may be regulated via an interaction between the V(H) promoter and 3' enhancer elements (here typified by the HS1,2 enhancer) in terminally differentiated B lineage cells. PMID- 10369418 TI - Regulation of the germline immunoglobulin Cgamma1 promoter by CD40 ligand and IL 4: dual role for tandem NF-kappaB binding sites. AB - Transcription of germline Ig constant region genes and associated switch regions is an early and essential step in heavy chain class switch recombination. Transcription of the germline Cgamma1 and C epsilon Ig genes is induced by IL-4 via STAT6 activation; CD40 signaling can independently induce transcription of these genes and act in synergy with IL-4 to increase expression. In the present study, we investigated the role of three tandem NF-kappaB sites (site 1, -95; site 2, -71; site 3, -53) in the regulation of the germline Cgamma1 Ig promoter by CD40 Ligand (CD40L) and IL-4 in the mouse B lymphoma cell line, BCL1-3B3. Germline gamma1 transcripts are induced by CD40L and by IL-4 in BCL1-3B3 and the combination of signals is synergistic, as in normal B cells. EMSA with crude nuclear extracts demonstrated that stimulation with CD40L results in the induction of NF-kappaB complexes that bind to each of the three NF-kappaB sites and are composed mainly of p50 and RelB, but also include c-Rel and p65. Surprisingly, site-specific mutagenesis of the NF-kappaB sites did not reduce CD40-responsiveness of germline gamma1 promoter-luciferase reporter constructs transiently transfected into BCL1-3B3. Mutation in any one NF-kappaB site, however, significantly reduced overall transcriptional activity of the promoter, both basal and induced, suggesting a role in basal promoter function. In addition, activation of the promoter by IL-4 was blocked by mutation of all three NF-kappaB sites and similarly reduced by mutation of site 1, suggesting that NF kappaB-STAT6 interactions may be necessary for STAT6-mediated transactivation of the germline gamma1 promoter. The results suggest that the three NF-kappaB sites may serve as a focus for formation of a higher-order transcription complex including STAT6, NF-kappaB and components of the basal transcription apparatus. PMID- 10369419 TI - Association of the interleukin-4 receptor alpha chain with p47phox, an activator of the phagocyte NADPH oxidase in B cells. AB - Interleukin (IL)-4 plays an important role in IgE synthesis in B cells and in Th2 differentiation in T cells. IL-4 conducts its biological activities through binding to the IL-4 receptor (IL-4R) on the surface of target cells. IL-4R are thought to be composed of the IL-4R alpha chain (IL-4R alpha) and either the IL 2R gamma chain or the IL-13R alpha chain. We have previously shown that the membrane-proximal portion in the cytoplasmic domain of the human IL-4R alpha (hIL 4R alpha) is critical for proliferation, generation of germline epsilon transcript, and activation of STAT6, based on analyses of truncated hIL-4R alphas. In this study, we found that p47phox, an activator of the phagocyte NADPH oxidase, binds to this portion by the two-hybrid system. Furthermore, we observed the association of p47phox with the hIL-4R alpha in B cells derived from a normal donor. These results suggest that p47phox is involved in the signal transduction of IL-4 in B cells. However, activation of STAT6, CD23 expression, and IgE synthesis induced by IL-4 were not affected in p47phox-deficient patients, which raises the possibility that p47phox may be important in other signaling activities as well in B cells. PMID- 10369420 TI - Analysis of human IgE epitope of Der f 2 with anti-Der f 2 mouse monoclonal antibodies. AB - Der f 2 is one of the major mite allergens recognized by human IgE antibodies of allergic patients. Using five anti-Der f 2 mouse monoclonal antibodies, human IgE epitopes of Der f 2 were analyzed. Among them, two monoclonal antibodies 15E11 and 13A4 inhibited the binding between Der f 2 and human IgE antibodies. To determine major IgE epitopes of Der f 2, epitopes for the monoclonal IgG antibodies were analyzed using 43 single site Der f 2 mutants constructed by site directed mutagenesis. Binding ability of 13A4 and 15E11 was decreased by the amino acid replacement around the C-terminus, and around 73rd, respectively. These results suggest that the C-terminal portion and the central portion around 73rd of Der f 2 were recognized by human IgE antibodies as major epitopes. The location of the putative IgE epitopes on 3-D structure of Der f 2 is also discussed. PMID- 10369421 TI - Generation and characterization of a novel single-gene-encoded single-chain immunoglobulin molecule with antigen binding activity and effector functions. AB - Monoclonal antibody (MAb) CC49 is a murine IgG1 that reacts with tumor-associated glycoprotein (TAG)-72, a pancarcinoma antigen. Clinical trials using radiolabeled CC49 for diagnostic imaging have demonstrated specific localization of more than 90% of carcinomas. The feasibility of adopting in vivo gene inoculation methods for antibody-based immunotherapy requires introduction and expression of two genes, encoding immunoglobulin (Ig) heavy and light chains, in a single cell to generate a functional antibody. To circumvent the problems inherent in this approach, we have constructed a single-gene encoding a single-chain immunoglobulin (SCIg) that, unlike previously developed SCIgs, contains all IgG domains. To construct the novel SCIg, the carboxyl end of the constant region of the chimeric (c) CC49 kappa chain is joined, via a 30 residue Gly-Ser linker peptide, to the amino terminus of the CC49 heavy chain. To our knowledge, neither a linker peptide this long nor a linkage between the constant light (C(L)) and variable heavy domains has been reported previously. Transfectomas developed by introducing the expression construct of the amplifiable gene in dihydrofolate reductase-deficient Chinese hamster ovary (CHO dhfr-) cells secrete a 160 kDa homodimeric molecule, SCIgcCC49. The in vitro antigen binding properties of SCIgcCC49 are comparable to those of cCC49 and SCIgcCC49deltaC(H)1, a single chain Ig deficient in constant heavy chain-1 (C(H)1) and C(L) domains. The antibody-dependent cellular cytotoxicity (ADCC) of SCIgcCC49 and cCC49 were also comparable. This single-gene approach for generating an immunoglobulin molecule may facilitate in vivo gene inoculation as well as ex vivo transfection of patients' cultured tumor-infiltrating lymphocytes for immunotherapy protocols for a variety of diseases, including cancer. PMID- 10369422 TI - Lymphoid specificity of a copy number-related expression of the H2-Kb transgene. AB - Transcriptional regulation of the MHC class I genes leading to their developmental and tissue specific expression is still poorly understood in spite of the recovery of a large variety of cis-controlling sequences and trans-acting factors pertaining to the 5' enhancer and the downstream regulatory element. Here we produced a series transgenic lines of mice with a genomic subclone of the H2 Kb gene consisting of 367 bp of the 5' upstream region, the coding region and 1.5 kb of the 3' downstream region and carrying all hitherto known regulatory sequences. The comparison of nine transgenic lines carrying the same H2-Kb transgene made it possible to ask whether the cis-information present in the transgene was sufficient for the tissue- and developmental-specific expression and its copy number dependence. We found the proper developmental onset of expression of the transgene at day 13 p.c. and correct tissue specific mRNA levels in adult mice. While in lymphoid tissues and in lung the number of transgene copies still correlated with RNA levels, the copy number dependence was completely lost in liver, kidney and embryonic tissues. Comparison with previously published H2-Kb transgenes indicates that the H2-Kb locus-controlling region is composed of more than one element. PMID- 10369423 TI - Inhibition of tumor necrosis factor-alpha with anti-diabetic agents. AB - It has recently been indicated that tumor necrosis factor-alpha (TNF-alpha) production is increased under chronic hyperglycemia and TNF-alpha has harmful effects on insulin sensitivity and possibly on chronic diabetic complications. Therefore it will be favorable for diabetes treatment if anti-diabetic agents also have anti-TNF-alpha activities. In this study, we have investigated effects of hypoglycemic sulfonylureas (gliclazide and glibenclamide) and a thiazolidinedione (troglitazone) on lipopolysaccharide-induced TNF-alpha production, which was evaluated by immunoassay and bioassay, in vivo using mice and partly in vitro using human peripheral blood mononuclear cells. Gliclazide significantly inhibited TNF-alpha production in vivo and also in vitro at a concentration of 10(-3) mol/l. However, glibenclamide had neither effect on TNF alpha production nor action. On the other hand, troglitazone inhibited action rather than production of TNF-alpha in vivo. In vitro troglitazone (10(-4) mol/l) significantly reduced cytolytic activity of TNF-alpha against LM cells. These results indicate that gliclazide and troglitazone have inhibitory effect on TNF alpha. PMID- 10369424 TI - Repaglinide versus glyburide: a one-year comparison trial. AB - This prospective, 1-year, multicenter, double-blind, randomized, parallel-group study was designed to show that repaglinide was at least equivalent to glyburide in patients with type 2 diabetes. Five hundred and seventy-six patients with type 2 diabetes of at least 6 months' duration were randomized to receive monotherapy with repaglinide (n = 383) or glyburide (n = 193). During weeks 1-8, doses were gradually increased to achieve a target fasting plasma glucose (FPG) range of 80 140 mg/dl. The final adjusted dose was maintained for 12 months. Repaglinide patients received a starting dose of 0.5 mg three times/day preprandially, adjusted as necessary to 1, 2 or 4 mg before breakfast, lunch and dinner. Glyburide patients received a starting dose of 2.5 mg before breakfast and placebo before lunch and dinner. Glyburide was increased as necessary to 5 or 10 mg before breakfast (placebo before lunch and dinner) or to 15 mg (10 mg before breakfast, placebo before lunch, and 5 mg before dinner). After study drug was stopped, patients were transferred to an appropriate therapy, as recommended by the investigator. Efficacy was assessed by changes from baseline in glycemic control parameters and in C-peptide, insulin, and lipid profiles. Repaglinide provided glycemic control that was at least as effective and potentially safer than that provided by glyburide. The glucose-lowering effect of repaglinide was most pronounced in pharmacotherapy-naive patients, who showed rapid and marked decreases in mean glycosylated hemoglobin levels from baseline (9.4%) to month 3 (7.6%) and month 12 (7.9%). Mean FPG levels also decreased overall in this group, from 222 mg/dl at baseline, to 175 mg/dl at month 3, to 188 mg/dl at month 12. At endpoint, morning C-peptide levels had increased significantly in glyburide treated patients compared with those treated with repaglinide, but morning fasting insulin levels did not differ significantly between the two groups. Repaglinide efficacy was sustained over 1 year and was not influenced by age or sex. Overall safety and changes in lipid profile and body weight were similar with both agents, with no significant change after extended pharmacotherapy. Weight gain data for the subset of pharmacotherapy-naive patients suggest that patients given repaglinide may gain less weight than those given glyburide. Repaglinide, at doses of 0.5-4.0 mg administered three times preprandially, was well tolerated and provided safe and consistently effectiveglycemic control during this 1-year study. Patients using repaglinide received the same therapeutic benefits as those using glyburide, and may have received additional benefits. PMID- 10369425 TI - Familial aggregation of diabetic kidney disease in Type 2 diabetes in south India. AB - The study was done to assess whether there was a familial aggregation of diabetic kidney disease (DKD) in Type 2 diabetic subjects. The profile of associated complications was also studied. Two groups of diabetic siblings of Type 2 diabetic patients, matched for age, body mass index (BMI) and duration of diabetes mellitus were studied. The siblings also had Type 2 diabetes. Group A comprised of siblings of probands with diabetic nephropathy and retinopathy (n = 30, M:F = 20:10) and Group B were siblings of probands without diabetic nephropathy or microalbuminuria (MAU) (n = 30, M:F = 14:16). Anthropometry, measurement of blood pressure and tests for proteinuria, MAU and retinopathy and ECG and biothesiometry were carried out for all study subjects. Persistent proteinuria was present in 15 (50%) siblings in group A and none in group B. MAU was detected in 26.7% (n = 7) in Group A and 3.3% (n = 1) in Group B (P = 0.057). Thus a total of 22 out of 30 cases in Group A had albuminuria. In Group A, seven (23.3%) had proteinuria and hypertension. Hypertension was present in nine (30.0%) in group A, and in five (16.7%) in group B (NS). Occurrence of retinopathy was found to be significantly higher in group A than in group B (33.3 vs 6.7%, chi2 = 5.1, P = 0.023). Abnormal ECG changes were present in 10% and 6.7% in Group A and Group B, respectively. In Group A, one patient had peripheral vascular disease (PVD) while in Group B none had PVD. A comparison of sib pairs, matched for age, duration of diabetes and the level of metabolic control showed that there was strong familial clustering of diabetic kidney disease in south Indians with Type 2 diabetes. This was independent of the familial clustering of diabetes. Prevalence of other vascular complications were also higher in Group A. PMID- 10369426 TI - Pulmonary diffusing capacity, serum angiotensin-converting enzyme activity and the angiotensin-converting enzyme gene in Japanese non-insulin-dependent diabetes mellitus patients. AB - We investigated the independent change in pulmonary diffusing capacity (DLCO) as one manifestation of pulmonary microangiopathy and to analyze the correlation between DLCO and serum ACE. We also examined the association between DLCO and the ACE genes. We examined pulmonary functions, especially %DLCO/VA (DLCO corrected by alveolar volume, percent predicted) in 54 NIDDM patients and 34 age-matched normal control subjects. Subjects were subdivided according to the degree of retinopathy. Serum ACE level was assayed by a colorimetric method in 54 patients and an insertion/deletion polymorphism in the ACE gene was amplified using the polymerase chain reaction in 52 of the 54 patients. There was a significant reduction of %DLCO/VA (percent predicted P < 0.05) in diabetic patients. In the proliferative retinopathy (PDR) group. %DLCO/VA was significantly (P < 0.05) lower than in the no diabetic retinopathy (NDR) and simple diabetic retinopathy (SDR) groups. Although the levels of serum ACE were within normal ranges in all diabetic groups, %DLCO/VA was negatively correlated with serum ACE values (r = 0.49, P < 0.0002, y = -1.4x + 109.3). Differences among DD, ID and II type of the ACE gene, with respect to the incidence of abnormal values of each clinical parameter, were not significant. DLCO was significantly reduced in patients with PDR and the serum ACE was significantly related to impaired DLCO. Our study suggests the existence of microangiopathic involvement of pulmonary vessels in NIDDM patients. PMID- 10369427 TI - Will acarbose improve the metabolic abnormalities of insulin-resistant type 2 diabetes mellitus? AB - Individuals with type 2 diabetes mellitus (n = 105; age 36-71 years) on diet therapy alone, and with quite good glycaemic control (mean HbA1c approximately 7.0%) were randomized to receive acarbose (100 mg three times daily) or placebo for 16 weeks, and changes in clinical and metabolic parameters indicative of Syndrome X were monitored. Fasting levels of glucose, glycosylated haemoglobin (HbA1c), true insulin, proinsulin, fibrinogen and lipids were measured four times weekly, and glucose, insulin, proinsulin and triglyceride responses to a standardized 1.6 MJ breakfast were determined at 0, 1 and 2 h post meal. Analysis was on an intention-to-treat basis. Fasting levels of glucose (P < 0.0001), triglycerides (P = 0.03) and HbA1c (P = 0.003) were reduced by acarbose over the 16 weeks of treatment. The mean change in HbA1c from week 0 to 16 differed by 0.4% (P = 0.003) between the two groups. Insulin (P = 0.06), proinsulin (P = 0.07) and glucose (P < 0.0001) responses to the standard meal were reduced. These data show that acarbose reduces fasting glucose and triglyceride levels, lowers HbA1c and limits the glycaemic and insulin response to food in individuals with type 2 diabetes mellitus with Syndrome X. Pharmacological agents that improve the metabolic environment and reduce insulin resistance have the potential to limit the progression of atherogenesis associated with type 2 diabetes mellitus. PMID- 10369428 TI - An inverse correlation between serum leptin levels and hemoglobin A1c in patients with non-insulin dependent diabetes mellitus. AB - We measured serum leptin concentrations in 70 patients with diabetes mellitus to investigate the relationship between serum leptin levels and glycemic control. A positive correlation between serum leptin levels and body mass index or plasma insulin was obtained as reported previously. The present study also demonstrated an inverse association of serum leptin levels with hemoglobin A1c (HbA1c). Multiple regression analysis revealed that HbA1c was an independent determinant of serum leptin levels. These results suggest that HbA1c may be a factor to influence serum leptin levels and that hyperglycemia for a long period or poorly controlled diabetes may reduce leptin levels. PMID- 10369429 TI - Long-term effect of epalrestat on cardiac autonomic neuropathy in subjects with non-insulin dependent diabetes mellitus. AB - To evaluate the effect of long-term administration of an aldose reductase inhibitor on diabetic cardiovascular autonomic neuropathy, 22 subjects with non insulin dependent diabetes mellitus (NIDDM, 11 men and 11 women, mean age; 64.8 +/- 7.8 years, duration of diabetes; 18.3 +/- 5.6 years) were administered epalrestat, one type of aldose reductase inhibitor, for 36 months. The changes in the coefficient of variation of the R-R interval (CV(R R)) during rest and the QTc interval were compared with 43 age-matched NIDDM (controls). During the study, the CV(R R) value gradually decreased in the controls, while it slightly increased in subjects treated with epalrestat. After 36 months, the CV(R R) value (2.31 +/- 1.09%) in subjects treated with epalrestat was significantly (P < 0.05) higher than that (1.84 +/- 0.75%) in the controls. There were no significant differences in QTc intervals in both groups. These results suggest that long-term administration of an aldose reductase inhibitor may be available for cardiac autonomic neuropathy in even relatively older diabetic subjects with long duration. PMID- 10369430 TI - Haemochromatosis gene mutations Cys282Tyr and His63Asp are not increased in Type 2 diabetic patients compared with the Canterbury (New Zealand) general population. AB - Genetic predisposition to haemochromatosis may be an important aetiological factor in some cases of Type 2 diabetes. Our aim was therefore to test the hypothesis that the haemochromatosis gene mutations Cys282Tyr and His63Asp are more prevalent in Type 2 diabetic patients compared with the Canterbury, New Zealand general population. We studied 230 consecutive patients referred to the Diabetes Services with age > or = 30 years and considered to have Type 2 diabetes. DNA was extracted from whole blood and amplified by polymerase chain reaction prior to restriction fragment length polymorphism analysis. The frequency of the mutations was compared with that observed previously in 1064 subjects from the Canterbury general population by chi2 testing. Iron was measured by a colorimetric method, transferrin by rate nephelometry and ferritin by immunoassay. There were 2/230 (0.8%) Cys282Tyr homozygous subjects in the diabetic group compared with 5/1064 (0.5%) NS in the general population. Although there was a trend to lower incidence of Cys282Tyr heterozygosity in the diabetic group, there was no significant difference for any of the six genotype frequencies between the two groups. Haemochromatosis gene mutations Cys282Tyr and His63Asp are therefore not increased in Type 2 diabetics compared with the general population. Transferrin saturation was a sensitive marker (100%) of genetic haemochromatosis, although ferritin had low specificity (77.8%). Genetic susceptibility to haemochromatosis is not an important aetiological factor for diabetes, and targeted screening of diabetic patients for haemochromatosis is not indicated. PMID- 10369431 TI - Early presentation of type 2 diabetes mellitus in young New Zealand Maori. AB - For 51 (5%) of the 1052 Maori patients registered with the Northland New Zealand (NZ) Diabetes Service, an initial diagnosis of diabetes was made before they had reached 30 years of age. We reviewed epidemiological and clinical data of this patient group. A total of 28 patients (55%) have type 2 diabetes, 18 (35%) have type 1 diabetes and for five patients, we were unable to determine the type of diabetes. The majority have positive family histories of diabetes (83%). The average age at diagnosis was 12.4 years for type 1 and 19.1 years for type 2 diabetes. Mean duration of diabetes and mean current HbA1c were not statistically different between the two groups, yet microalbuminuria or nephropathy was present in 62% of type 2 patients, but only 18% of the type 1 group. These statistics show that a significant number of Northland NZ Maori develop type 2 diabetes at an earlier age than expected and have a high incidence of renal complications. These findings contrast with previous New Zealand studies on the incidence of diabetes in young Maori, but are similar to those of recent overseas studies of ethnic groups with a high incidence of diabetes. PMID- 10369432 TI - Power, control and resistance in the timing of health and care. AB - In the modern period, time has been commodified and can be bought, sold and bartered. This paper argues that this means that while time may be a tool of power and control, it is also a site for resistance to power. Four case studies are evaluated from this perspective. A day-case surgery unit is examined to identify the routinization of health care, with time as a tool of control. Decisions over discharge from hospital following surgery demonstrate how time may be 'sold' to 'buy' future time. The disruption of routines during surgery suggest how time may be used as a means of resisting power, while in residential homes, old people find time on their hands as it is transformed into something which must be filled. It is argued that resistance is not achieved by recourse to 'natural' (as opposed to cultural) time, but by rethinking time creatively. PMID- 10369433 TI - Spatiality, embodiment and hazards encountered in the making of pots. AB - The aim is to relate embodiment to more distant spatiality. The vehicle is an ethnographic study in one pottery factory over a total of two and a half years of how potters encounter hazards to health and safety at work. Potters' experiences of the body and space differ from the body and space as imagined by medicine, the law and capital. Though these experiences are relatively coherent, in the wider context they carry little legitimacy and are seldom expressed. Each of three traditions, that of Marx, represented by H. Lefebvre, that of Weber and also Foucault, by B. Turner and that of Durkheim, by M. Douglas, provides a template for organising a different theme in the field notes: (1) the dialectic of real, imagined and lived spatialities, (2) the production of varied embodiment in employment and (3) the symbolic meanings of lived space. A link is made between spatiality and embodiment by considering what happens when the rhythms of manual work break down and ill-health or injury comes about. This particular link is given theoretical expression by developing parallels between Lefebvre's dialectic of spatiality and the dialectic of disease, illness and sickness, as discussed by Young and Frankenberg. PMID- 10369434 TI - Family structure and patient survival in an African-American end-stage renal disease population: a preliminary investigation. AB - Few studies have examined the influence of the family on the course of chronic illness in African-Americans. We explore the relationship between family structure, defined as marital status and household composition, and patient survival. Patient gender was examined as a possible moderator in this relationship. Using data from a survey of 476 African-American end-stage renal disease (ESRD) patients, a significant association between household composition and patient survival was found. Results from Cox proportional hazards model, controlling for patient age, indicated that patients who live in 'complex' households (i.e. those with a partner and/or others) are at greater risk for shortened survival as compared to those who live alone or with a spouse/partner (p < 0.05). When we examined whether patient gender moderates this relationship, female patients who live in these households were found to be at 2 times greater risk for shortened survival (p < 0.01) than female patients who live alone or with their spouse/ partner only. Family structure was not significantly associated with survival in male patients. Discussion and implications of findings are addressed. PMID- 10369435 TI - Information needs in terminal illness. AB - Despite evidence that doctor-patient communication affects important patient outcomes, patient expectations are often not met. Communication is especially important in terminal illness, when the appropriate course of action may depend more on patient values than on medical dogma. We sought to describe the issues important to terminally ill patients receiving palliative care and to determine whether patient characteristics influence the needs of these patients. We utilized a multimethod approach, first conducting interviews with 22 terminally ill individuals, then using these data to develop a more structured instrument which was administered to a second population of 56 terminally ill patients. Patient needs and concerns were described and associations between patient characteristics and issues of importance were evaluated. Seven key issues were identified in the initial interviews: change in functional status or activity level; role change; symptoms, especially pain; stress of the illness on family members; loss of control; financial burden and conflict between wanting to know what is going on and fearing bad news. Overall, respondent needs were both disease- and illness-oriented. Few easily identifiable patient characteristics were associated with expressed concerns or needs, suggesting that physicians need to individually assess patient needs. Terminally ill patients receiving palliative care had needs that were broad in scope. Given that few patient characteristics predicted responses, and that the majority opinion may not accurately reflect that of an individual patient, health care providers must be aware of the diverse concerns among this population and individualize assessment of each patient's needs and expectations. PMID- 10369436 TI - Psychosocial problem disclosure by primary care patients. AB - The vast majority of psychologically distressed primary care patients present exclusively somatic concerns at the outsets of their visits. However, it is not known how often such patients subsequently disclose psychosocial problems to their primary care physicians (PCPs) and what variables predict such disclosures. Our objectives were to measure, among psychologically distressed primary care patients, the frequency of disclosure of psychosocial problems (disclosure), the effects of prior psychosocial inquiry (prior inquiry) by PCPs and various patient variables on disclosure, and the effect of disclosure on mental health problem recognition (recognition) by PCPs. The study was based in the practices of 69 community-based PCPs and involved 308 adult patients with 28-item General Health Questionnaire scores of 5 or greater, indicating significant psychological distress. Disclosure occurred during 51% of visits overall and 67% of visits with prior inquiry. The odds of disclosure were increased by prior inquiry (p < 0.001), greater physician-patient familiarity (p < 0.001) and greater severity of patient psychological distress (p < 0.001). Prior inquiry and physician-patient familiarity had a negative interaction (p < 0.05) of smaller size than either variable's main effect, so that their combined effect on disclosure exceeded the effect of either variable alone but was less than multiplicative. The estimated odds ratio for recognition given disclosure was 24.13 (95% confidence interval, 11.28-51.63) after adjustment for the effects of significant covariates. We conclude that if PCPs inquire, most psychologically distressed, somatically presenting patients will disclose psychosocial problems. Inquiry is particularly productive with unfamiliar patients. PCPs can engender a substantial increase in psychosocial disclosure simply by adding one or two questions about mood or interpersonal problems to their clinical interviews. PMID- 10369437 TI - Gender, psychosocial factors and the use of medical services: a longitudinal analysis. AB - Many researchers have reported gender differences in levels of reported symptoms, morbidity, mortality and medical care utilization, but the debate continues about the underlying causes of these differences. Some have argued that women use more medical services because they are more sensitive to symptoms and interested in health, while others believe that women's greater service utilization arises from the fact that women experience more morbidities than do men. To date, these questions have not been studied prospectively. Using data from a household interview survey carried out in 1970-1971 and linked to 22 years of health services utilization records, we explored the effects of gender, self-reported health status, mental and physical symptom levels, health knowledge, illness behaviors and health concerns and interest on the long-term use of health services. After controlling for the aforementioned factors, female gender remained an independent predictor of higher utilization over the 22-year period studied, and psychosocial and health factors measured at the initial interview predicted service use even 19-22 years later. Controlling for factors identified as likely causes of gender-related differences in healthcare utilization, gender remains an important predictor of medical care use before and after removing sex specific utilization. In addition, the consistent predictive ability of attitudinal and behavioral factors, combined with the finding that health knowledge did not predict utilization, indicates that efforts to help patients assess their service needs should target the attitudinal and behavioral factors that vary with gender, rather than health-related knowledge alone. PMID- 10369438 TI - Psychosocial outcome assessments for use in cardiac rehabilitation service evaluation: a 10-year systematic review. AB - A variety of measures are currently used to assess psychosocial outcome (quality of life) in cardiac rehabilitation programmes. However, there is no consensus on the most appropriate instruments to use. Instruments that are not sufficiently responsive to change in cardiac populations are unsuitable as audit tools as they underrepresent the benefits of programme attendance. To identify the most responsive instruments in cardiac rehabilitation populations a systematic overview of studies for the 10-year period 1986-1995 was conducted. The following databases were searched: Medline, Psychlit, Cinahl and Sociofile and 32 relevant studies were identified. The effect size statistic (a comparison of the magnitude of change to the variability in baseline scores) was used to determine those instruments most responsive to change. The following instruments were identified as being responsive in more than one study: Beck Depression Inventory, Global Mood Scale, Health Complaints Checklist, Heart Patients Psychological Questionnaire and Speilberger State Anxiety Inventory. There is little consensus on psychosocial evaluation instrument use in the cardiac rehabilitation literature. A number of measures show significant potential for routine outcome assessment. Formal assessment of these instruments is recommended to inform final recommendations about instrument selection for audit and evaluation purposes in cardiac rehabilitation. PMID- 10369439 TI - Disability terminology in the media: a comparison of newspaper reports in Canada and Israel. AB - The terminology used to refer to persons with disability may both reflect and influence attitudes towards them. Negative references may perpetuate negative attitudes and stereotypes. This is of particular importance in the mass media which reaches a broad spectrum of the population. This study looked at disability terminology used in major newspapers in Canada and Israel. It focused on the nature of that terminology and whether its use was related to other factors, such as the disability model reflected in the article, the content of the article (e.g. attributes of the disabled person) and its context (e.g. type of newspaper, feature versus news items). Overall, the use of inappropriate terminology of varying types was quite prevalent in both countries. In addition, in Canada there were a considerable number of articles which had no direct reference to the disability. In general, the terminology used was considerably more positive in articles dealing with individual persons with disabilities (as opposed to groups), with disabled children and with problems of mobility and rights. The results of the study indicate that the choice of terminology cannot be explained by journalistic expedience and conciseness alone. PMID- 10369440 TI - The 'dop' system, alcohol abuse and social control amongst farm workers in South Africa: a public health challenge. AB - Many farm workers in South Africa continue to live and work under adverse conditions that are the legacy of apartheid policies. Despite its official prohibition, the arrangement by which workers are given alcohol' as a benefit of employment, known as the 'dop' system, appears to persist. Even though it is a minority of farms that currently actively practice the dop system, the ramifications of the historical 'institutionalisation of massive alcohol consumption are widespread. Heavy alcohol consumption is not only directly injurious to the health of farm workers and their families, but places them at risk to various social and environmental hazards. This is illustrated in a case of pesticide poisoning in which 24 workers were poisoned when given wine contaminated with the carbamate insecticide aldicarb. The case illustrates (i) the ongoing application of the dop system on farms in South Africa and (ii) the interaction between social factors and chemical exposures amongst farm workers. Public perceptions about the natural tendencies of 'coloured' people to drink heavily have much to do with perpetuating the dop system, and reinforcing a system geared towards the social control of rural farm workers and their families. The dop system poses a major challenge to the public health authorities in South Africa who are charged with the task of restructuring health services to address the human rights and health needs of marginal farming communities within a primary health care framework. PMID- 10369441 TI - The impact of market oriented reforms on choice and information: a case study of cataract surgery in outer London and Stockholm. AB - In the early 1990s, a set of market-oriented reforms was introduced into health care systems of the UK and Sweden, two exemplary cases of reliance on planned budgeting and integrated provision of services. In the pursuit of increased efficiency, several County Councils in Sweden have followed the public competition model, while in the UK internal market reforms were introduced. It was expected that the separation of functions of planners and purchasers from those of providers, which were to be freely chosen by the former, would achieve higher allocative efficiency but also enhance users' satisfaction with care. This paper uses cataract surgery as a case study to trace the impact of competition among providers on choice and information. Qualitative research methods were employed to record the perception of changes in their type and amount as it was given to both purchasers and patients. A set of open ended and standardised questionnaires was designed to elicit the views of all actors involved and to measure the likely transformations. Four study sites from Outer London were selected representing the diversity of responses, and the only existing large provider of eye services to Stockholm County Council was used. The analysis of the data showed that the quasi-market reforms have resulted in a change of attitude of providers. Some improvements in the amount and type of information given to purchasers and patients could also be detected, although as far as direct users were concerned, the demand has not been fully satisfied. However, the impact on choice available to patients and purchasers alike seemed to be adverse, an effect that was particularly strong in the UK case. PMID- 10369442 TI - Demograghic and socio-economic determinants of community and hospital services costs for people with HIV/AIDS in London. AB - We examined the influence of demographic, social and economic background of people with HIV/AIDS in London on total community and hospital services costs. This was a retrospective study of community and hospital service use, needs and costs based on structured questionnaires administered by trained interviewers and costing information obtained from the service purchasers and providers, based on two Genito-urinary Medicine clinics in London: the Jefferiss Wing at St. Mary's Hospital and Patric Clements at the Central Middlesex Hospital, London, England. The subjects were 225 HIV infected patients (105 asymptomatic, 59 symptomatic non AIDS and 61 AIDS). We found that over and above well established determinants of health care costs for HIV infected people such as disease stage and transmission category, social and economic factors such as employment and support of a living in partner significantly reduced community services costs. Private health insurance had a similar effect, though only a small proportion of HIV people had such cover. The cost of community services for HIV infected non-European Union nationals, mainly of African origin, was one quarter that for the European Union nationals. Community services costs were highest for heterosexually infected women and lowest for heterosexually infected men after adjusting for other factors. Hospital services costs were significantly higher for HIV infected people lacking educational qualifications and employment. We conclude that access to community care for HIV infected non-EU nationals appears to be very poor as the cost of their community services was one quarter that for the EU nationals after adjusting for the effects of transmission category, disease stage, living with a partner, employment and having a private health insurance. Additional incentives for informal care for HIV infected people could be a cost-effective way to improve their community health service provisions. PMID- 10369444 TI - A critique of seven assumptions behind psychological trauma programmes in war affected areas. AB - Programmes costing millions of dollars to address 'posttraumatic stress' in war zones have been increasingly prominent in humanitarian aid operations, backed by UNICEF, WHO, European Community Humanitarian Office and many nongovernmental organisations. The assumptions underpinning this work, which this paper critiques with particular reference to Bosnia and Rwanda, reflect a globalisation of Western cultural trends towards the medicalisation of distress and the rise of psychological therapies. This paper argues that for the vast majority of survivors posttraumatic stress is a pseudocondition, a reframing of the understandable suffering of war as a technical problem to which short-term technical solutions like counselling are applicable. These concepts aggrandise the Western agencies and their 'experts' who from afar define the condition and bring the cure. There is no evidence that war-affected populations are seeking these imported approaches, which appear to ignore their own traditions, meaning systems, and active priorities. One basic question in humanitarian operations is: whose knowledge is privileged and who has the power to define the problem? What is fundamental is the role of a social world, invariably targeted in today's 'total' war and yet still embodying the collective capacity of survivor populations to mourn, endure and rebuild. PMID- 10369443 TI - Doing a shotgun: a drug use practice and its relationship to sexual behaviors and infection risk. AB - There has been a rise in the frequency with which inhalational routes such as smoking are used for illicit drug use. A growing population of new inhalational drug users augments the pool of individuals at risk for transition to injection drug use. Further, illicit drug smoking has been implicated in the transmission of a variety of pathogens by the respiratory route, and crack smoking has been associated with an increased risk of HIV infection, particularly through the exchange of high-risk sex for drugs. Shotguns are an illicit drug smoking practice in which smoked drugs are exhaled or blown by one user into the mouth of another user. We conducted a series of ethnographic observations to attempt to characterize more fully the practice of shotgunning, the range of associated behaviors, and the settings and contexts in which this practice occurs. Shotguns may be seen as a form of drug use which has close ties to sexual behaviors, and which has both pragmatic and interpersonal motivations, combining in a single phenomenon the potential direct and indirect risk of disease transmission by sexual, blood borne and respiratory routes. These data support the need to develop and evaluate comprehensive risk reduction interventions, which take into consideration the relationships between interpersonal and sexual behaviors and specific forms of drug use. PMID- 10369445 TI - 'With the best of reasons': cervical cancer prevention policy and the suppression of sexual risk factor information. AB - Cervical cancer is a very common but largely preventable cancer. Despite considerable medical knowledge of risk and even causal factors, possible social behavioural strategies for the primary prevention of cervical cancer have rarely been explored as a viable addition to cervical screening. We examine key policy documents and interview 18 key informants on cervical cancer prevention in New Zealand. Using a discourse analytic approach we identify and discuss two discourses (which we have labelled 'protectionism' and 'right to know') which inform positions on whether or not women should be provided with information regarding sexual risk factors for cervical cancer. Cervical cancer prevention policy in New Zealand, which largely reflects a protectionist discourse, suppresses sexual risk factor information and focuses exclusively on cervical screening. The right to know discourse informs an alternative position, which contends that women have a right to be informed about risk factors. We discuss these positions in relation to questions about women's rights, the principle of informed choice, and attempts to judge what is in women's 'best interests.' PMID- 10369446 TI - Political self-characterization of US women physicians. AB - The political self-characterization of US physicians, especially including women physicians, has been poorly described. We used data from the 4,501 respondents to the Women Physicians' Health Study (WPHS), a stratified random sample of US women M.D.s surveyed in 1993-1994, to assess US women physicians' political characteristics. US women physicians were most likely to consider themselves politically moderate (36.6% of respondents). More considered themselves liberal (28.4%) or very liberal (8.8%) than considered themselves conservative (20.5%) or very conservative (5.8%). US women physicians predominantly bring moderate and liberal voices to political discourse. Organizations that wish to attract US women physician members should consider promoting less conservative policies. PMID- 10369447 TI - Geographies of health perception in Quebec: a multilevel perspective. AB - Self perceived health is a widely used measure and, in Quebec, it has been shown to vary significantly between geographical areas. In the present study, these geographical variations are examined in a multilevel analysis in order to disentangle compositional (individual characteristics) and contextual (place) effects. The analysis recognizes four levels of variation: individual, household, local and regional. Similar analyses carried out in Britain, have considered only two levels: individual and local. Data come from the 1992-1993 Quebec Health and Social Survey, a general household survey using a stratified two-stage sampling design. Health perception (the response variable) is considered with a set of individual predictor characteristics reflecting gender, lifestyle, socio-economic conditions, marital status and social support. Results show the existence of significant local area variations in health perception after having allowed for individual characteristics and variations at the household level. At the regional level, however, no systematic and significant variations remain although some individual regions are found to have a significant impact on health perception. PMID- 10369448 TI - The persisting effect of unemployment on health and social well-being in men early in working life. AB - In our studies of the effects of unemployment in the early working life of men in a British national birth cohort we have shown elsewhere that this experience was part of a longer term accumulation of social and health disadvantage. This present study asks whether men's unemployment also inflicted potential longterm damage to future socio-economic chances and health. We therefore constructed indicators of socio-economic circumstances and health at 33 years from factors already shown to be associated with health in later life. For the socio-economic indicator we used a combination of income, occupational status and home ownership and described this as socio-economic capital. For the health indicator we combined scores of body mass index, leisure time exercise, frequency of eating fresh fruit and of smoking, and described this as health capital. After controlling for pre-labour market socio-economic and health factors, prolonged unemployment is shown here to reduce significantly both socio-economic and health capital by age 33 years. We conclude that the experience of prolonged unemployment early in the working life of this population of young men looks likely to have a persisting effect on their future health and socio-economic circumstances. PMID- 10369449 TI - Effects of oral propylthiouracil treatment on nitric oxide production in rat aorta. AB - The effects of oral propylthiouracil (PTU) treatment on vascular nitric oxide (NO) production were studied in the rat aorta. Rats were fed a standard low fat diet with or without 0.1% PTU, for 2 or 4 weeks, or for 2 weeks with additional thyroxine injections. Concentration response curves were then constructed to phenylephrine (PE) in both endothelium-intact and denuded aortic rings from these animals and after incubation with 0.1 mM L-N(G)nitroarginine (L-NOARG). In addition, expression of nitric oxide synthase (NOS) was analysed in sections of aorta from PTU-treated and control rats using rabbit polyclonal antibodies to both inducible NOS (iNOS) and endothelial NOS (eNOS). Oral PTU treatment resulted in a significant reduction in both the maximum response (control, 0.53+/-0.02; 2 week PTU, 0.20+/-0.07; 4 week PTU, 0.07+/-0.02 g mg(-1)) and vessel sensitivity (EC50 values: control, 9.10x10(-8)+/-0.67; 2 week PTU, 7.45x10(-7)+/-1.15; 4 week PTU, 9.73x10(-7)+/-0.45 M) to PE in endothelium-intact vessel rings, as compared to controls (P<0.05). Both endothelial removal and incubation with L-NOARG restored the maximum response after 2, but not 4 weeks, although, in general, vessel sensitivity was not altered by either treatment. Vessels from PTU-treated rats given thyroxine injections showed no significant differences between any of the dose response curve parameters. Immunohistochemical analysis suggested that labelling for eNOS may be increased after PTU treatment as compared to control animals, whereas iNOS antibody immunoreactivity was not different between the two groups. These results suggest that the hyporesponsiveness to PE observed after oral PTU treatment is, in part, due to enhanced nitric oxide (NO) production by the endothelium, and demonstrate for the first time that thyroid hormones may play a role in the regulation of eNOS activity in the rat aorta. PMID- 10369450 TI - Molecular analysis of the Na+ channel blocking actions of the novel class I anti arrhythmic agent RSD 921. AB - RSD 921 is a novel, structurally unique, class I Na+ channel blocking drug under development as a local anaesthetic agent and possibly for the treatment of cardiac arrhythmias. The effects of RSD 921 on wild-type heart, skeletal muscle, neuronal and non-inactivating IFMQ3 mutant neuronal Na+ channels expressed in Xenopus laevis oocytes were examined using a two-electrode voltage clamp. RSD 921 produced similarly potent tonic block of all three wild-type channel isoforms, with EC50 values between 35 and 47 microM, whereas the EC50 for block of the IFMQ3 mutant channel was 110+5.5 microM. Block of Na+ channels by RSD 921 was concentration and use-dependent, with marked frequency-dependent block of heart channels and mild frequency-dependent block of skeletal muscle, wild-type neuronal and IFMQ3 mutant channels. RSD 921 produced a minimal hyperpolarizing shift in the steady-state voltage-dependence of inactivation of all three wild type channel isoforms. Open channel block of the IFMQ3 mutant channel was best fit with a first order blocking scheme with k(on) equal to 0.11+/-0.012x10(6) M( 1) s(-1) and k(off) equal to 12.5+/-2.5 s(-1), resulting in KD of 117+/-31 microM. Recovery from open channel block occurred with a time constant of 14+/ 2.7 s(-1). These results suggest that RSD 921 preferentially interacts with the open state of the Na+ channel, and that the drug may produce potent local anaesthetic or anti-arrhythmic action under conditions of shortened action potentials, such as during anoxia or ischaemia. PMID- 10369451 TI - Pharmacological analysis of the novel, selective alpha1-adrenoceptor antagonist, KMD-3213, and its suitability as a tritiated radioligand. AB - Pharmacological profiles of tritiated KMD-3213, a new antagonist of alpha1 adrenoceptor (AR), were examined in recombinant and native alpha1-AR, and compared with those of prazosin (PZ) and tamsulosin (YM-617). In saturation experiments, [3H]-KMD (10-2000 pM) showed high affinity for alpha1a-AR (pK(D) = 10.5). However, no significant binding to alpha1b-AR and insufficient/unsaturated binding to alpha1d-AR were observed at concentrations up to 2000 pM. In contrast, [3H]-PZ and [3H]-YM bound to all subtypes with high affinity (pK(D)>9). In competition experiments, KMD-3213 also had higher affinity for alpha1a-AR than for other two subtypes; pKi = 10.4, 8.1 and 8.6 for alpha1a-, alpha1b- and alpha1d-AR, respectively. [3H]-KMD also bound to the native alpha1A-AR (rat submaxillary gland) with high affinity, but not to alpha1B-AR (rat liver). In rat kidney which expresses alpha1A- and alpha1B-AR, [3H]-KMD and [3H]-PZ bound to a single high-affinity site (pK(D) = 10.8 and 10.1, respectively) with distinct amount of binding sites (Bmax = 159 and 267 fmol mg(-1) protein, respectively). [3H]-PZ binding sites consisted of low- and high-affinity sites for KMD-3213 (pKi = 7.6 and 10.7, respectively), for WB4101 (pK = 8.1 and 10.0) and for YM-617 (pKi = 8.7 and 10.8). The proportion of the high affinity site was approximately 60% in these drugs which was compatible to the ratio between Bmax of [3H]-KMD and [3H]-PZ. [3H]-KMD binding sites consisted of a single site for these drugs with affinities which were similar to those of the high affinity sites in [3H]-PZ binding. In functional experiments, KMD-3213 antagonized the contractile responses to NS-49 or noradrenaline (NA) with higher affinity in functional alpha1A- (rat caudal artery, pA2= 10.0 against NS-49) and alpha1L-AR (dog mesenteric artery, pA2 = 9.9 against NA) than in alpha1B- (dog carotid artery, pA2 = 7.7 against NA) and alpha1D-AR (rat thoracic aorta, pA2 = 8.3 against NA). These results confirm the alpha1A-AR selectivity and high affinity of KMD-3213, and indicate that [3H]-KMD can label selectively alpha1A-AR. PMID- 10369452 TI - Potassium ions and endothelium-derived hyperpolarizing factor in guinea-pig carotid and porcine coronary arteries. AB - Experiments were designed to determine in two arteries (the guinea-pig carotid and the porcine coronary arteries) whether or not the endothelium-derived hyperpolarizing factor (EDHF) can be identified as potassium ions, and to determine whether or not the inwardly rectifying potassium current and the Na+/K+ pump are involved in the hyperpolarization mediated by EDHF. The membrane potential of vascular smooth muscle cells was recorded with intracellular microelectrodes in the presence of N(omega)-L-nitro-arginine (L-NA) and indomethacin. In vascular smooth muscle cells of guinea-pig carotid and porcine coronary arteries, acetylcholine and bradykinin induced endothelium-dependent hyperpolarizations (-18+/-1 mV, n = 39 and -19+/-1 mV, n = 7, respectively). The hyperpolarizations were not affected significantly by ouabain (1 microM), barium chloride (up to 100 microM) or the combination of ouabain plus barium. In both arteries, increasing extracellular potassium concentration by 5 or 10 mM induced either depolarization or in a very few cases small hyperpolarizations which never exceeded 2 mV. In isolated smooth muscle cells of the guinea-pig carotid artery, patch-clamp experiments shows that only 20% of the vascular smooth muscle cells expressed inwardly rectifying potassium channels. The current density recorded was low (0.5+/-0.1 pA pF(-1), n = 8). These results indicate that, in two different vascular preparations, barium sensitive-inwardly rectifying potassium conductance and the ouabain sensitive-Na+/K+ pump are not involved in the EDHF mediated hyperpolarization. Furthermore, potassium did not mimic the effect of EDHF pointing out that potassium and EDHF are not the same entity in those arteries. PMID- 10369453 TI - The effect of moxonidine on feeding and body fat in obese Zucker rats: role of hypothalamic NPY neurones. AB - The antihypertensive agent moxonidine, an imidazoline Ii-receptor agonist, also induces hypophagia and lowers body weight in the obese spontaneously hypertensive rat, but the central mediation of this action and the neuronal pathways that moxonidine may interact with are not known. We studied whether moxonidine has anti-obesity effects in the genetically-obese and insulin-resistant fa/fa Zucker rat, and whether these are mediated through inhibition of the hypothalamic neuropeptide Y (NPY) neurones. Lean and obese Zucker rats were given moxonidine (3 mg kg(-1) day(-1)) or saline by gavage for 21 days. Moxonidine decreased food intake throughout by 20% in obese rats (P<0.001) and by 8% in lean rats (P<0.001), and reduced weight gain that final body weight was 15% lower in obese (P<0.001) and 7% lower in lean (P<0.01) rats than their untreated controls. Plasma insulin and leptin levels were decreased in moxonidine-treated obese rats (P<0.01 and P<0.05), but unchanged in treated lean rats. Uncoupling protein-1 gene expression in brown adipose tissue was stimulated by 40-50% (P< or =0.05) in both obese and lean animals given moxonidine. Obese animals given moxonidine showed a 37% reduction in hypothalamic NPY mRNA levels (P = 0.01), together with significantly increased NPY concentrations in the paraventricular nucleus (P<0.05), but no changes in the arcuate nucleus or other nuclei; this is consistent with reduced NPY synthesis in the arcuate nucleus and blocked release of NPY in the paraventricular nucleus. In lean animals, moxonidine did not affect NPY levels or NPY mRNA. The hypophagic, thermogenic and anti-obesity effects of moxonidine in obese Zucker rats may be partly due to inhibition of the NPY neurones, whose inappropriate overactivity may underlie obesity in this model. PMID- 10369454 TI - Role of blood-brain barrier P-glycoprotein in limiting brain accumulation and sedative side-effects of asimadoline, a peripherally acting analgaesic drug. AB - Studies with knockout mice lacking mdr1a P-glycoprotein (P-gp) have previously shown that blood-brain barrier P-gp is important in preventing the accumulation of several drugs in the brain. Asimadoline (EMD 61753) is a peripherally selective kappa-opioid receptor agonist which is under development as a therapeutic analgaesic. From the structural characteristics of this drug and its peripheral selectivity, we hypothesized that it is transported by P-gp. Using a pig-kidney polarized epithelial cell line transfected with mdr cDNAs, we demonstrate that asimadoline is transported by the mouse mdr1a P-gp and the human MDR1 P-gp. Furthermore, we show that in mdr1a/1b double knockout mice, the absence of P-gp leads to a 9 fold increased accumulation of asimadoline in the brain. In line with this accumulation difference, mdr1a/1b (-/-) mice are at least 8 fold more sensitive to the sedative effect of asimadoline than wild-type mice. Interestingly, the oral uptake of asimadoline was not substantially altered in mdr1a/1b (-/-) mice. Our results demonstrate that for some drugs, P-gp in the blood-brain barrier can have a therapeutically beneficial effect by limiting brain penetration, whereas at the same time intestinal P-gp is not a significant impediment to oral uptake of the drug. PMID- 10369455 TI - Cross-talk between group IIA-phospholipase A2 and inducible NO-synthase in rat renal mesangial cells. AB - Features of glomerulonephritis are expression of the inducible form of NO synthase (iNOS) as well as expression of the secretory group IIA-phospholipase A2 (sPLA2) in mesangial cells. Interleukin 1beta (IL-1beta) induces both enzymes with a similar time course resulting in an increase in nitrite production and sPLA2-IIA activity. In this study we investigated the relationship between the formation of NO and sPLA2-IIA induction in rat renal mesangial cells. Incubation of mesangial cells with the NO-donor, spermine-NONOate, for 24 h induced sPLA2 IIA mRNA expression and activity, whereas S-nitroso glutathione alone had only a small stimulatory effect. Stimulation of cells with IL-1beta caused a marked increase in sPLA2-IIA mRNA and activity that were potentiated 3 fold by both NO donors. Coincubation of cells with IL-1beta and the NOS inhibitor, L-N(G) monomethylarginine (L-NMMA), caused a dose-dependent inhibition of cytokine induced sPLA2-IIA mRNA expression and activity. sPLA2-IIA activity was not stimulated by 8-bromo-cyclic GMP indicating that NO-induced sPLA2-IIA induction is independent of cyclic GMP-mediated signal transduction. These data show that NO contributes to the expression by cytokines of sPLA2-IIA and establishes a novel type of interaction between iNOS and sPLA2-IIA in mesangial cells. This cross-talk between inflammatory mediators may help to promote and sustain an inflammatory state in the kidney. PMID- 10369456 TI - Antagonistic action of pitrazepin on human and rat GABA(A) receptors. AB - Pitrazepin, 3-(piperazinyl-1)-9H-dibenz(c,f) triazolo(4,5-a)azepin is a piperazine antagonist of GABA in a variety of electrophysiological and in vitro binding studies involving GABA and glycine receptors. In the present study we have investigated the effects of pitrazepin, and the GABA(A) antagonist bicuculline, on membrane currents elicited by GABA in Xenopus oocytes injected with rat cerebral cortex mRNA or cDNAs encoding alpha1beta2 or alpha1beta2gamma2s human GABA(A) receptor subunits. The three types of GABA(A) receptors expressed were reversibly antagonized by bicuculline and pitrazepin in a concentration dependent manner. GABA dose-current response curves for the three types of receptors were shifted to the right, in a parallel manner, by increasing concentrations of pitrazepin. Schild analyses gave pA2 values of 6.42+/-0.62, n = 4, 6.41+/-1.2, n = 5 and 6.21+/-1.24, n = 6, in oocytes expressing rat cerebral cortex, alpha1beta2 or alpha1beta2gamma2s human GABA(A) receptors respectively (values are given as means +/- s.e. mean), and the Hill coefficients were all close to unity. All this is consistent with the notion that pitrazepin acts as a competitive antagonist of these GABA(A) receptors; and that their antagonism by pitrazepin is not strongly dependent on the subunit composition of the receptors here studied. Since pitrazepin has been reported to act also at the benzodiazepine binding site, we studied the effect of the benzodiazepine antagonist Ro 15-1788 (flumazenil) on the inhibition of alpha1beta2gamma2s receptors by pitrazepin. Co-application of Ro 15-1788 did not alter the inhibiting effect of pitrazepin. Moreover, pitrazepin did not antagonize the potentiation of GABA-currents by flunitrazepam. All this suggests that pitrazepin does not affect the GABA receptor-chloride channel by interacting with the benzodiazepine receptor site. PMID- 10369457 TI - Characterization of the cyclic nucleotide phosphodiesterase subtypes involved in the regulation of the L-type Ca2+ current in rat ventricular myocytes. AB - The effects of several phosphodiesterase (PDE) inhibitors on the L-type Ca current (I(Ca)) and intracellular cyclic AMP concentration ([cAMP]i) were examined in isolated rat ventricular myocytes. The presence of mRNA transcripts encoding for the different cardiac PDE subtypes was confirmed by RT-PCR. IBMX (100 microM), a broad-spectrum PDE inhibitor, increased basal I(Ca) by 120% and [cAMP]i by 70%, similarly to a saturating concentration of the beta-adrenoceptor agonist isoprenaline (1 microM). However, MIMX (1 microM), a PDE1 inhibitor, EHNA (10 microM), a PDE2 inhibitor, cilostamide (0.1 microM), a PDE3 inhibitor, or Ro20-1724 (0.1 microM), a PDE4 inhibitor, had no effect on basal I(Ca) and little stimulatory effects on [cAMP]i (20-30%). Each selective PDE inhibitor was then tested in the presence of another inhibitor to examine whether a concomitant inhibition of two PDE subtypes had any effect on I(Ca) or [cAMP]i. While all combinations tested significantly increased [cAMP]i (40-50%), only cilostamide (0.1 microM)+ Ro20-1724 (0.1 microM) produced a significant stimulation of I(Ca) (50%). Addition of EHNA (10 microM) to this mix increased I(Ca) to 110% and [cAMP]i to 70% above basal, i.e. to similar levels as obtained with IBMX (100 microM) or isoprenaline (1 microM). When tested on top of a sub-maximal concentration of isoprenaline (1 nM), which increased I(Ca) by (approximately 40% and had negligible effect on [cAMP]i, each selective PDE inhibitor induced a clear stimulation of [cAMP]i and an additional increase in I(Ca). Maximal effects on I(Ca) were approximately 8% for MIMX (3 microM), approximately 20% for EHNA (1 3 microM), approximately 30% for cilostamide (0.3-1 microM) and approximately 50% for Ro20-1724 (0.1 microM). Our results demonstrate that PDE1-4 subtypes regulate I(Ca) in rat ventricular myocytes. While PDE3 and PDE4 are the dominant PDE subtypes involved in the regulation of basal I(Ca), all four PDE subtypes determine the response of I(Ca) to a stimulus activating cyclic AMP production, with the rank order of potency PDE4>PDE3>PDE2>PDE1. PMID- 10369458 TI - Distinct effects of ceramide-generating pathways in prostate adenocarcinoma cells. AB - Ceramide, generated by the hydrolysis of sphingomyelin, mediates the actions of several cytokines such as tumour necrosis factor-alpha (TNF-alpha) interferon gamma and interleukin-1beta (IL-1beta), including their inhibitory effect on tumour proliferation. We have evaluated the role of ceramide in the proliferation of prostate cancer by using the human prostate adenocarcinoma LNCaP cell line. Treatment of LNCaP cells with neutral or acidic sphingomyelinase or addition of C8- or C2-ceramide, two cell permeable analogues of endogenous ceramide, induced a profound inhibition of cell proliferation. This effect appeared after 24 h, was still present after 72 h of exposure to the drugs and exhibited concentration dependency (10-200 and 5-200 mU ml(-1) for neutral and acidic sphingomyelinase, respectively, and 1-25 microM for C8-ceramide). The inhibitory effect on cell growth caused by neutral sphingomyelinase and ceramides was rapidly reversible as LNCaP cells rapidly regained their previous proliferation rate following withdrawal of the treatment. IL-1beta produced profound inhibition of LNCaP cell proliferation and caused enhanced ceramide formation. No clear features of apoptotic cell death were detectable by either oligonucleosome formation, cytofluorimetric analysis or nuclear staining following exposure of LNCaP cells to neutral sphingomyelinase, ceramide or IL-1beta. However, clear changes in LNCaP cell cycle distribution were detectable following these treatments. In contrast, treatment with acidic sphingomyelinase or TNF-alpha induced apoptotic death detectable by flow cytometric analysis and bisbenzimide staining. In conclusion, our data demonstrate that preferential activation of distinct enzymatic pathways by cytokines may lead to different outcomes in the viability of LNCaP cells. PMID- 10369459 TI - Gi-Protein alpha-subunit mRNA antisense oligonucleotide inhibition of Gi-coupled receptor contractile activity in the epididymis of the guinea-pig. AB - We have used a reversible permeabilization method to facilitate the entry of Gialpha1, 2 and 3 G-protein subunit mRNA antisense or mismatch oligonucleotides into intact tissue, to investigate the G-protein alpha-subunit coupling of alpha2 adrenoceptors, neuropeptide Y (NPY) Y1, and A1 adenosine receptors in preparations of the epididymis of the guinea-pig. The alpha2-adrenoceptor agonist, xylazine, elicited concentration dependent contractions from preparations of phenylephrine (3 microM)-stimulated epididymis (pEC50 value 6.52+/-0.39, maximum response 236+/-41 mg force). Compared to respective mismatch controls the incubation of preparations with Gialpha2, but not with Gialpha1 or Gialpha3 mRNA antisense oligonucleotides (30 microM) reduced the maximal xylazine potentiation of phenylephrine (3 microM)-stimulated contractility (to 51+/-12% of Gialpha2 mismatch control). The oligonucleotide incubations had no effect upon the pEC50 values of xylazine. The A1 adenosine receptor agonist, cyclopentyladenosine (CPA) elicited concentration dependent contractions from preparations of phenylephrine (3 microM)-stimulated epididymis (pEC50 value 7.66+/-0.57, maximum response 208+/-54 mg force). Incubation of preparations of epididymis with Gialpha1, but neither Gialpha2 nor Gialpha3 antisense oligonucleotides reduced the maximal CPA-potentiation of phenylephrine (3 microM) stimulated contractions (to 55+/-17% of Gialpha1 mismatch control), pEC50 values were not affected. The incubation of preparations with Gialpha2 antisense mRNA oligonucleotides reduced the maximal NPY-potentiation of phenylephrine (3 microM) stimulated contractions (to 62+/-15% of Gialpha mismatch control). Compared with Gialpha2 mismatch controls, the incubation of preparations with Gialpha1 and Gialpha3 oligonucleotides also reduced the NPY-potentiation of phenylephrine (3 microM)-stimulated contractions. These studies indicate that, in the guinea-pig epididymis, alpha2-adrenoceptors and A1 adenosine receptors preferentially couple to effectors through Gialpha2 and Gialpha1 subunits respectively. In contrast NPY receptors may elicit effects through either Gialpha1, 2 or 3 subunits. PMID- 10369460 TI - Quantitative autoradiographic studies of relaxin binding in rat atria, uterus and cerebral cortex: characterization and effects of oestrogen treatment. AB - The binding characteristics of the relaxin receptor in rat atria, uterus and cortex were studied using a [33P]-labelled human gene 2 relaxin (B33) and quantitative receptor autoradiography. The binding kinetics of [33P]-human gene 2 relaxin (B33) were investigated in slide-mounted rat atrial sections. The binding achieved equilibrium after 60 min incubation at room temperature (23+/-1 degrees C) and dissociated slowly. The association and dissociation rate constants were 4.31+/-0.34x10(8) M(-1) x min(-1) and 1.55+/-0.38x10(-3) min(-1) respectively. Thus, the kinetic dissociation constant was 3.46+/-0.59 pM. Binding was saturable to a single population of non-interacting sites throughout atria, in uterine myometrium and the 5th layer of cerebral cortex. The binding affinities (pK(D)) of [33P]-human gene 2 relaxin (B33) were 8.92+/-0.09 in atrial myocardium and 8.79+/-0.04 in cerebral cortex of male rats, and 8.79+/-0.10 in uterine myometrium. Receptor densities in the cerebral cortex and atria were higher than in uterine myometrium, indicating that relaxin also has important roles in non reproductive tissues. In male rats, treatment with 17beta-oestradiol (20 microg in 0.1 ml sesame oil s.c., 18-24 h) significantly decreased the density of relaxin receptors in atria and cerebral cortex. Identical treatment in female rats had no significant effect in atria and cerebral cortex, but it significantly increased the density of relaxin receptors in uterine myometrium. Relaxin binding was competitively displaced by porcine and rat native relaxins. Porcine native relaxin binds to the relaxin receptor in male rat atria (8.90+/-0.02), and cerebral cortex (8.90+/-0.03) and uterine myometrium (8.89+/-0.03) with affinities not significantly different from human gene 2 (B33) relaxin. Nevertheless, rat relaxin binds to the receptors with affinities (8.35+/-0.09 in atria, 8.22+/-0.07 in cerebral cortex and 8.48+/-0.06 in uterine myometrium) significantly less than human gene 2 (B33) and porcine relaxins. Quantitative receptor autoradiography is the method of choice for measurement of affinities and densities of relaxin receptor in atria, uterine myometrium and cerebral cortex. High densities were found in all these tissues. 17beta-oestradiol treatment produced complex effects where it increased the densities of relaxin receptors in uterus but decreased those in atria and cerebral cortex of the male rats, and had no effect on the atria and cerebral cortex of the female rats. PMID- 10369461 TI - Heterogeneity of prejunctional NPY receptor-mediated inhibition of cardiac neurotransmission. AB - Neuropeptide Y (NPY) has been proposed as the candidate inhibitory peptide mediating interactions between sympathetic and vagal neurotransmission in several species, including man. Here, we have defined the NPY receptors involved in modulation of cardiac autonomic neurotransmission using receptor-selective agonists and antagonists in the rabbit and guinea-pig isolated right atria. In isolated atrial preparations, sympathetically-mediated tachycardia (ST; with atropine 1 microM) or vagally-mediated bradycardia (VB; with propranolol 0.1-1 microM) in response to electrical field stimulation (EFS, 1-4 pulses) were tested 0-30 min after incubation with single concentrations of vehicle, NPY (0.01-10 microM), the Y2 receptor agonist N-Acetyl-[Leu28,31]NPY(24-36) (termed N A[L]NPY(24-36)) or the Y1 receptor agonist [Leu31,Pro34]NPY (LP). The effect of NPY on the concentration-chronotropic response curves to isoprenaline and bethanechol were also assessed. Guinea-pig atria: NPY and N-A[L]NPY(24-36) caused concentration-dependent inhibition of VB and ST to EFS. Both peptides caused maximal inhibition of VB and ST within 10 min incubation and this remained constant. LP caused a concentration-dependent, transient inhibition of ST which was antagonized by the Y1-receptor antagonist GR231118 (0.3 microM), with apparent competitive kinetics. Rabbit atria: NPY (1 or 10 microM) had no effect on VB at any time point, but both NPY and LP caused a transient (approximately 10 min) inhibition of sympathetic tachycardia. This inhibition could be prevented by 0.3 microM GR231118. N-A[L]NPY(24-36) had no effect on ST. NPY had no effect on the response to beta-adrenoceptor stimulation by isoprenaline nor muscarinic receptor stimulation by bethanechol in either species. Thus, in the guinea-pig, NPY causes a stable inhibition of both VB and ST to EFS via Y2 receptors and transient inhibition of ST via Y1 receptors. In contrast in the rabbit, NPY has no effect on the cardiac vagus and prejunctional inhibition of ST is transient and mediated by a Y1-like receptor (rather than Y2). Therefore it would be surprising if NPY plays a functional role in modulation of cardiac neurotransmission in the rabbit. PMID- 10369462 TI - Effects of GABA on noradrenaline release and vasoconstriction induced by renal nerve stimulation in isolated perfused rat kidney. AB - We examined effects of gamma-aminobutyric acid (GABA) on vasoconstriction and noradrenaline (NA) release induced by electrical renal nerve stimulation (RNS) in the isolated pump-perfused rat kidney. RNS (1 and 2 Hz for 2.5 min each, 0.5-ms duration, supramaximal voltage) increased renal perfusion pressure (PP) and renal NA efflux. GABA (3, 10 and 100 microM) attenuated the RNS-induced increases in PP by 10-40% (P<0.01) and NA efflux by 10-30% (P<0.01). GABA did not affect exogenous NA (40 and 60 nM)-induced increases in PP. The selective GABA(B) agonist baclofen (3, 10 and 100 microM) also attenuated the RNS-induced increases in PP and NA efflux, whereas the RNS-induced responses were relatively resistant to the selective GABA(A) agonist muscimol (3, 10 and 100 microM). The selective GABA(B) antagonist 2-hydroxysaclofen (50 microM), but not the selective GABA(A) antagonist bicuculline (50 microM), abolished the inhibitory effects of GABA (10 microM) on the RNS-induced responses. The selective alpha2-adrenoceptor antagonist rauwolscine (10 nM) enhanced the RNS-induced responses. GABA (3, 10 and 100 microM) potently attenuated the RNS-induced increases in PP by 40-60% (P<0.01) and NA efflux by 20-50% (P<0.01) in the presence of rauwolscine. Prazosin (10 and 30 nM) suppressed the RNS-induced increases in PP by about 70 80%. Neither rauwolscine (10 nM) nor GABA (10 microM) suppressed the residual prazosin-resistant PP response. These results suggest that GABA suppresses sympathetic neurotransmitter release via presynaptic GABA(B) receptors, and thereby attenuates adrenergically induced vasoconstriction in the rat kidney. PMID- 10369463 TI - Minor structural changes in nicotinoid insecticides confer differential subtype selectivity for mammalian nicotinic acetylcholine receptors. AB - The major nitroimine insecticide imidacloprid (IMI) and the nicotinic analgesics epibatidine and ABT-594 contain the 6-chloro-3-pyridinyl moiety important for high activity and/or selectivity. ABT-594 has considerable nicotinic acetylcholine receptor (AChR) subtype specificity which might carry over to the chloropyridinyl insecticides. This study considers nine IMI analogues for selectivity in binding to immuno-isolated alpha1, alpha3 and alpha7 containing nicotinic AChRs and to purported alpha4beta2 nicotinic AChRs. Alpha1- and alpha3 containing nicotinic AChRs (both immuno-isolated by mAb 35, from Torpedo and human neuroblastoma SH-SY5Y cells, respectively) are between two and four times more sensitive to DN-IMI than to (-)-nicotine. With immuno-isolated alpha3 nicotinic AChRs, the tetrahydropyrimidine analogues of IMI with imine or nitromethylene substituents are 3-4 fold less active than (-)-nicotine. The structure-activity profile with alpha3 nicotinic AChRs from binding assays is faithfully reproduced in agonist potency as induction of 86rubidium ion efflux in intact cells. Alpha7-containing nicotinic AChRs of SH-SY5Y cells (immuno-isolated by mAb 306) and rat brain membranes show maximum sensitivity to the tetrahydropyrimidine analogue of IMI with the nitromethylene substituent. The purported alpha4beta2 nicotinic AChRs [mouse (Chao & Casida, 1997) and rat brain] are similar in sensitivity to DN-IMI, the tetrahydropyrimidine nitromethylene and nicotine. The commercial insecticides (IMI, acetamiprid and nitenpyram) have low to moderate potency at the alpha3 and purported alpha4beta2 nicotinic AChRs and are essentially inactive at alpha1 and alpha7 nicotinic AChRs. In conclusion, the toxicity of the analogues and metabolites of nicotinoid insecticides in mammals may involve action at multiple receptor subtypes with selectivity conferred by minor structural changes. PMID- 10369464 TI - Comparison of the effects of [Phe1psi(CH2-NH)Gly2]nociceptin(1-13)NH2 in rat brain, rat vas deferens and CHO cells expressing recombinant human nociceptin receptors. AB - Nociceptin(NC) is the endogenous ligand for the opioid receptor like-1 receptor (NC-receptor). [Phe1(psi)(CH2-NH)Gly2]Nociceptin(1-13)NH2 ([F/G]NC(1-13)NH2) has been reported to antagonize NC actions in peripheral guinea-pig and mouse tissues. In this study, we investigated the effects of a range of NC C-terminal truncated fragments and [F/G]NC(1-13)NH2 on NC receptor binding, glutamate release from rat cerebrocortical slices (rCX), inhibition of cyclic AMP accumulation in CHO cells expressing the NC receptor (CHO(NCR)) and electrically evoked contractions of the rat vas deferens (rVD). In radioligand binding assays, a range of ligands inhibited [125I]-Tyr14-NC binding in membranes from rCX and CHO(NCR) cells. As the peptide was truncated there was a general decline in pKi. [F/G]NC(1-13)NH2 was as potent as NC(1-13)NH2. The order of potency for NC fragments to inhibit cyclic AMP accumulation in whole CHO(NCR) cells was NCNH2> or =NC=NC(1-13)NH2>NC(1-12)NH2> >NC(1-11)NH2. [F/G]NC(1-13)NH2 was a full agonist with a pEC50 value of 8.65. NCNH2 and [F/G]NC(1-13)NH2 both inhibited K+ evoked glutamate release from rCX with pEC50 and maximum inhibition of 8.16, 48.5+/-4.9% and 7.39, 58.9+/-6.8% respectively. In rVD NC inhibited electrically evoked contractions with a pEC50 of 6.63. Although [F/G]NC(1-13)NH2, displayed a small (instrinsic activity alpha = 0.19) but consistent residual agonist activity, it acted as a competitive antagonist (pA2 6.76) in the rVD. The differences between [F/G]NC(1-13)NH2 action on central and peripheral NC signalling could be explained if [F/G]NC(1-13)NH2 was a partial agonist with high strength of coupling in the CNS and low in the periphery. An alternative explanation could be the existence of central and peripheral receptor isoforms. PMID- 10369465 TI - Concentration-dependent isoflurane effects on depolarization-evoked glutamate and GABA outflows from mouse brain slices. AB - The synaptic concentrations of glutamate and gamma-aminobutyric acid (GABA) are modulated by their release and re-uptake. The effects of general anaesthetics on these two processes remain unclear. This study evaluates the effects of isoflurane, a clinically important anaesthetic, on glutamate and GABA release and re-uptake in superfused mouse cerebrocortical slices. Experiments consisted of two 1.5-min exposures to 40 mM KCl in 30 min intervals. During the second exposure, different concentrations of isoflurane with and without 0.3 mM L transpyrrolidine-2,4-dicarboxylic acid (PDC, a competitive inhibitor of glutamate uptake transporter) or 1 mM nipecotic acid (a competitive inhibitor of GABA uptake transporter) were introduced. The ratios of the second to first KCl-evoked increases in glutamate and GABA were used to determine the isoflurane concentration-response curves. The results can be described as a sum of two independent processes, corresponding to the inhibitions of release and re-uptake, respectively. The EC50 values for the inhibitions of release and re-uptake were 295+/-16 and 805+/-43 microM for glutamate, and 229+/-13 and 520+/-25 microM for GABA, respectively. Addition of PDC did not significantly affect glutamate release but shifted the re-uptake curve to the left (EC50= 315+/-20 microM). Nipecotic acid completely blocked GABA uptake, rendering isoflurane inhibition of GABA re-uptake undetectable. Our data suggest that isoflurane inhibits both the release and re-uptake of neurotransmitters and that the inhibitions occur at different EC50's. For GABA, both EC50's are within the clinical concentration range. The net anaesthetic effect on extracellular concentrations of neurotransmitters, particularly GABA, depends on the competition between inhibition of release and that of re-uptake. PMID- 10369466 TI - Effect of the neuroprotective compound SR57746A on nerve growth factor synthesis in cultured astrocytes from neonatal rat cortex. AB - The neurotrophic factor promoting activity of the neuroprotective compound SR57746A was evaluated in primary cultures of neonatal rat cortical astrocytes by studying the synthesis of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF). A concentration- and time-dependent increase of nerve growth factor mRNA was induced by SR57746A (10 nM-1 microM). In these astrocytes, BDNF mRNA contents were increased to a significant but smaller extent, and beta actin mRNA showed no variation. SR57746A (1 microM) induced increases of both de novo protein translation after 6 h of incubation and NGF release into the extracellular medium after 6-24 h. These effects were preceded by a transient augmentation of junB, c-fos and c-jun mRNA contents. These increases of AP-1 family mRNA were associated with increased nuclear AP-1 binding activity. The results show that SR57746A can increase the synthesis and release of NGF in rat cortical astrocytes. Such effects may contribute to the drug's previously described neuroprotective effects. PMID- 10369467 TI - Effects of (-)-tertatolol, (-)-penbutolol and (+/-)-pindolol in combination with paroxetine on presynaptic 5-HT function: an in vivo microdialysis and electrophysiological study. AB - The antidepressant efficacy of selective serotonin reuptake inhibitors (SSRIs) might be enhanced by co-administration of 5-HT1A receptor antagonists. Thus, we have recently shown that the selective 5-HT1A receptor antagonist, WAY 100635, blocks the inhibitory effect of an SSRI on 5-HT cell firing, and enhances its ability to elevate extracellular 5-HT in the forebrain. Here we determined whether the beta-adrenoceptor/5-HT1A receptor ligands (+/-)-pindolol, (-) tertatolol and (-)-penbutolol, interact with paroxetine in a similar manner. Both (-)-tertatolol (2.4 mg kg(-1) i.v.) and (-)-penbutolol (2.4 mg kg(-1) i.v.) enhanced the effect of paroxetine (0.8 mg kg(-1) i.v.) on extracellular 5-HT in the frontal cortex, whilst (+/-)-pindolol (4 mg kg(-1) i.v.) did not. (-) Tertatolol (2.4 mg kg(-1) i.v.) alone caused a slight increase in 5-HT however, ( )-penbutolol (2.4 mg kg(-1) i.v.) alone had no effect. In electrophysiological studies (-)-tertatolol (2.4 mg kg(-1) i.v.) alone had no effect on 5-HT cell firing but blocked the inhibitory effect of paroxetine. In contrast, (-) penbutolol (0.1-0.8 mg kg(-1) i.v.) itself inhibited 5-HT cell firing, and this effect was reversed by WAY 100635 (0.1 mg kg(-1) i.v.). We have recently shown that (+/-)-pindolol inhibits 5-HT cell firing via a WAY 100635-sensitive mechanism. Our data suggest that (-)-tertatolol enhances the effect of paroxetine on forebrain 5-HT via blockade of 5-HT1A autoreceptors which mediate paroxetine induced inhibition of 5-HT cell firing. In comparison, the mechanisms by which ( )-penbutolol enhances the effect of paroxetine on extracellular 5-HT is unclear, since (-)-penbutolol itself appears to have agonist properties at the 5-HT1A autoreceptor. Indeed, the agonist action of (+/-)-pindolol at 5-HT1A autoreceptors probably explains its inability to enhance the effect of paroxetine on 5-HT in the frontal cortex. Overall, our data suggest that both (-)-tertatolol and (-)-penbutolol are superior to (+/-)-pindolol in terms of enhancing the effect of an SSRI on extracellular 5-HT. Both (-)-tertatolol and (-)-penbutolol are worthy of investigation for use as adjuncts to SSRIs in the treatment of major depression. PMID- 10369468 TI - ET(A) receptors are the primary mediators of myofilament calcium sensitization induced by ET-1 in rat pulmonary artery smooth muscle: a tyrosine kinase independent pathway. AB - We have investigated the possibility that ET-1 can induce an increase in myofilament calcium sensitivity in pulmonary artery smooth muscle. Arterial rings were permeabilized using alpha-toxin (120 microg ml(-1)), in the presence of A23187 (10 microM) to 'knock out' Ca2+ stores, and pre-constricted with pCa 6.8 (buffered with 10 mM EGTA). In the presence of this fixed Ca2+ concentration, 1 microM ET-1 induced a sustained, reversible constriction of 0.15 mN. Pulmonary arterial rings were freeze-clamped at the peak of the induced constriction (time matched). Subsequent densitometric analysis revealed that ET-1 (1 microM) increased the level of phosphorylated myosin light chains by 34% compared to an 11% increase in the presence of pCa 6.8 alone. In contrast to ET-1, the selective ET(B) receptor agonist Sarafotoxin S6C (100 nM) failed to induce a significant constriction. The constriction induced by 1 microM ET-1 was reversibly inhibited when the preparation was preincubated (15 min) with the ETA receptor antagonist BQ 123 (100 microM). The constriction measured 0.13 mN in the absence and 0.07 mN in the presence of 100 microM BQ 123. In contrast, the constriction induced by 1 microM ET-1 measured 0.19 mN in the absence and 0.175 mN following a 15 min pre incubation with the ET(B) antagonist BQ 788 (100 microM). The constriction induced by 1 microM ET-1 measured 0.14 mN in the presence and 0.13 mN following pre-incubation with the tyrosine kinase inhibitor Tyrphostin A23 (100 microM). We conclude that ET-1 induced an increase in myofilament calcium sensitivity in rat pulmonary arteries via the activation of ET(A) receptors and by a mechanism(s) independent of tyrosine kinase. PMID- 10369469 TI - AMP is a partial agonist at the sheep cardiac ryanodine receptor. AB - We have investigated the ability of AMP to modulate the native sheep cardiac ryanodine receptor (RyR) channel at various cytosolic [Ca2+]. Channels were incorporated into planar phospholipid bilayers and current fluctuations through the bilayer were monitored under voltage clamp conditions. We demonstrate that AMP only exhibits agonist activity if the cytosolic [Ca2+] is sufficiently high. Even in the presence of a high cytosolic [Ca2+] (65 microM), AMP cannot fully open the channel and the maximum open probability (Po) observed is approximately 0.3 at 2 mM AMP. Concentrations of AMP above the maximally activating level cause inactivation of the channel. Our experiments indicate that AMP is an agonist with such low efficacy at the ATP sites on the cardiac RyR that it is effectively an antagonist of ATP-induced increases in Po. Our study demonstrates that the number of phosphates attached to the 5'-carbon of the ribose ring of adenine-based compounds determines the efficacy of the ligand to increase the Po of the cardiac RyR. Substitution of groups at this position may lead to the identification of potent antagonists at ATP sites on RyR. PMID- 10369470 TI - Effects of hydroxocobalamin and carboxy-PTIO on nitrergic transmission in porcine anococcygeus and retractor penis muscles. AB - The effects of carboxy-PTIO and hydroxocobalamin were studied on nitrergic transmission in anococcygeus and retractor penis muscles taken during post mortem examination from young male pigs. In both muscles under resting conditions, electrical field stimulation (EFS) caused contractions that were sensitive to tetrodotoxin (1 microM) and were greatly inhibited by prazosin (1 microM) and guanethidine (10-30 microM), but were not significantly affected by atropine (1 microM). In the anococcygeus muscle, but not in the retractor penis muscle, guanethidine produced a prolonged contraction. After tone was raised by guanethidine in the anococcygeus or by phenylephrine (1 microM) in the presence of guanethidine in the retractor penis, EFS caused tetrodotoxin-sensitive relaxations. The EFS-induced relaxations were abolished by the NO synthase inhibitor N(G)-L-nitro-arginine methyl ester (L-NAME; 100 microM) and its effect was partly overcome by L-arginine (1 mM), indicating it was mediated by nitrergic nerves. Carboxy-PTIO (0.1-1 mM) had no significant effect in reducing stimulation induced nitrergic relaxations in either muscle. However, hydroxocobalamin (0.1-1 mM) caused concentration-dependent reductions of nitrergic relaxations in both muscles. Relaxations to exogenous nitric oxide (1 microM) in both muscles were abolished by carboxy-PTIO (0.3 mM) and hydroxocobalamin (0.1 mM). There were no differences in reactivity to carboxy-PTIO or hydroxocobalamin between anococcygeus and retractor penis muscles from the same species (pig). The finding also confirms earlier observations that the nitrergic transmitter is generally resistant to the NO-scavenger carboxy-PTIO. PMID- 10369471 TI - 2-Phenyl-imidazo[1,2-a]pyridine derivatives as ligands for peripheral benzodiazepine receptors: stimulation of neurosteroid synthesis and anticonflict action in rats. AB - Selective activation of peripheral benzodiazepine receptors (PBRs) in adrenal cells and brain oligodendrocytes promotes steroidogenesis. Three 2-phenyl imidazo[1,2-a]pyridine derivatives (CB 34, CB 50 and CB 54) have now been investigated with regard to their selectivity for PBRs and their ability to stimulate central and peripheral steroidogenesis in rats. The three CB compounds (10(-10)-10(-4) M) potently inhibited the binding of the PBR ligand [3H]-PK 11195 to brain and ovary membranes in vitro, without substantially affecting [3H] flunitrazepam binding to central benzodiazepine receptors. These compounds (10( 7)-10(-4) M) also had little or no marked effects on GABA-evoked Cl- currents in voltage-clamped Xenopus oocytes expressing human alpha1beta2gamma2S GABA(A) receptors. In addition, they failed to affect ligands binding to GABA(B), D1/D2 dopamine, muscarinic acetylcholine, N-methyl-D-aspartic acid and opiate receptors. Intraperitoneal administration of CB compounds (3-50 mg kg(-1)) induced a dose-dependent increase in the concentrations of neuroactive steroids in plasma and brain. The brain concentrations of pregnenolone, progesterone, allopregnanolone and allotetrahydrodeoxycorticosterone (THDOC) showed maximal increases in 96+/-3, 126+/-14, 110+/-12 and 70+/-13% above control, respectively, 30 to 60 min after injection of CB 34 (25 mg kg(-1)). CB 34 also increased the brain concentrations of neuroactive steroids in adrenalectomized-orchiectomized rats, although to a lesser extent than in sham-operated animals, suggesting that CB compounds stimulate brain steroidogenesis independently of their effects on peripheral tissues. The increase in brain and plasma neurosteroid content induced by CB 34 was associated with a marked anticonflict effect in the Vogel test. Our results indicate that the three CB compounds tested are specific and potent agonists at peripheral benzodiazepine receptors, and that they stimulate steroidogenesis in both the brain and periphery. PMID- 10369472 TI - Adenosine A3 receptors on human eosinophils mediate inhibition of degranulation and superoxide anion release. AB - The role of adenosine A3 receptors on human eosinophil degranulation and superoxide anion (O2-) release was studied in vitro using the complement fragment C5a as the main stimulus and employing a number of selective agonists and antagonists. In the presence of cytochalasin B (CB), C5a induced a dose-dependent release of the granular eosinophil peroxidase (EPO), but not O2-, whereas in the absence of CB O2- , but not EPO, was released. C5a-induced EPO release was inhibited dose-dependently by the selective A3 agonist N6-(3-iodobenzyl)-5'-N methylcarbamoyladenosine (IB-MECA) and to a lesser extent by the less-selective N6-2-(4-amino-3-iodophenyl) ethyladenosine (APNEA). The IC50 (95% CI) for IB-MECA was 6.8 microM (3.1-12.0 microM). At concentrations up to 100 microM, neither adenosine nor the selective A1 agonist N-cyclopentyladenosine (CPA) and the selective A2 agonist 2-[[2-[4-(2-carboxyethyl)phenyl]ethyl]amino]-N ethylcarboxamidoadenosine (CGS 21680) had any significant effect. The inhibitory effect of IB-MECA was almost completely abolished by pre-treatment with 1 microM of the selective A3 antagonist 9-chloro-2-(2-furyl)-5 phenylactylamino[1,2,4]triazolo[1,5-c]quina zoline (MRS 1220), but not the selective A1 antagonist 1,3-dipropyly-8-cyclopentylxanthine (DPCPX) or the selective A2 antagonist 3,7-dimethyl-1-propargylxanthine (DMPX). IB-MECA also significantly inhibited C5a-induced O2- release with IC50 (95% CI) of 9.5 microM (4.6-13.1 microM) whereas adenosine and the A1 agonist CPA potentiated this effect at low concentrations. The potentiation appeared to be a result of their direct O2- release from these cells, probably mediated via A1 receptors. The inhibition by IB-MECA was selectively reversed by MRS 1220. These results show that the A3 receptors on human eosinophils mediate inhibition of both degranulation and O2- release and suggest a therapeutic potential for A3 agonists in diseases such as asthma in which activated eosinophils are involved. PMID- 10369473 TI - Effects of the soluble guanylyl cyclase activator, YC-1, on vascular tone, cyclic GMP levels and phosphodiesterase activity. AB - The vasomotor and cyclic GMP-elevating activity of YC-1, a novel NO-independent activator of soluble guanylyl cyclase (sGC), was studied in isolated rabbit aortic rings and compared to that of the NO donor compounds sodium nitroprusside (SNP) and NOC 18. Similarly to SNP and NOC 18, YC-1 (0.3-300 microM) caused a concentration-dependent, endothelium-independent relaxation that was greatly reduced by the sGC inhibitor 1-H-[1,2,4]oxadiazole[4,3-a]quinoxalin-1-one (ODQ 10 microM; 59% inhibition of dilation induced by 100 microM YC-1) suggesting the activation of sGC as one mechanism of action. Preincubation with YC-1 (3 and 30 microM) significantly increased the maximal dilator responses mediated by endogenous NO in aortic rings that was released upon exposure to acetylcholine, and enhanced the dilator response to the exogenous NO-donors, SNP and NOC 18, by almost two orders of magnitude. Vasoactivity induced by SNP and YC-1 displayed different kinetics as evidenced by a longlasting inhibition by YC-1 (300 microM) on the phenylephrine (PE)-induced contractile response, which was not fully reversible even after extensive washout (150 min) of YC-1, and was accompanied by a long-lasting elevation of intracellular cyclic GMP content. In contrast, SNP (30 microM) had no effect on the vasoconstrictor potency of PE, and increases in intravascular cyclic GMP levels were readily reversed after washout of this NO donor compound. Surprisingly, YC-1 not only activated sGC, but also affected cyclic GMP metabolism, as it inhibited both cyclic GMP break down in aortic extracts and the activity of phosphodiesterase isoforms 1-5 in vitro. In conclusion, YC-1 caused persistent elevation of intravascular cyclic GMP levels in vivo by activating sGC and inhibiting cyclic GMP break down. Thus, YC-1 is a highly effective vasodilator compound with a prolonged duration of action, and mechanisms that are unprecedented for any previously known sGC activator. PMID- 10369474 TI - Prostaglandin DP receptors positively coupled to adenylyl cyclase in embryonic bovine tracheal (EBTr) cells: pharmacological characterization using agonists and antagonists. AB - Various prostaglandin agonists representing various classes of receptor subtypes were evaluated for their ability to stimulate adenylyl cyclase via the endogenous DP receptor in embryonic bovine tracheal (EBTr) cells. Two antagonists were used to block the agonist-induced cyclic AMP production. ZK118182 (EC50 = 16+/-4 nM), RS-93520 (EC50 = 23+/- 4 nM), SQ27986 (EC50 = 33+/-9 nM), ZK110841 (EC50 = 33+/-5 nM), BW245C (EC50 = 59+/-19 nM) and PGD2 (EC50=101+/-10 nM) (n = 4-70) were the most potent agonists. Whilst most compounds were full agonists (Emax = 100% relative to PGD2), BW245C was significantly more efficacious than PGD2 (Emax = 121+/-3%; P<0.001) and RS-93520 appeared to be a partial agonist (Emax = 64+/-9%; P<0.001). Agonists from the EP (e.g. enprostil; misoprostol; butaprost), FP (e.g. cloprostenol; fluprostenol; PHXA85), IP (iloprost; PGI2) and TP (U46619) prostanoid receptor classes were weak agonists or inactive in the EBTr cell assay system. The DP-receptor antagonist, BWA868C, showed a competitive antagonist profile with pA2 values of 8.00+/-0.02 and 8.14+/-0.13 in Schild analyses with two structurally different agonists, BW245C and ZK118182, respectively (n = 3). AH6809, another purported DP-receptor antagonist, weakly inhibited PGD2- and ZK 18182-induced cyclic AMP production (K(i)s = 808+/-193 nM and 782+/-178 nM, respectively). The current studies have characterized the DP receptor positively coupled to adenylyl cyclase in EBTr cells using a wide range of agonist and antagonist prostaglandins. These data support the utility of the EBTr cell line as a useful tool for the evaluation of DP receptor agonists and antagonists and for profiling other classes of prostaglandins. PMID- 10369475 TI - Comparison of antagonist potencies at pre- and post-synaptic GABA(B) receptors at inhibitory synapses in the CA1 region of the rat hippocampus. AB - Synaptic activation of gamma-aminobutyric acid (GABA)B receptors at GABA synapses causes (a) postsynaptic hyperpolarization mediating a slow inhibitory postsynaptic potential/current (IPSP/C) and (b) presynaptic inhibition of GABA release which depresses IPSPs and leads to paired-pulse widening of excitatory postsynaptic potentials (EPSPs). To address whether these effects are mediated by pharmacologically identical receptors the effects of six GABA(B) receptor antagonists of widely ranging potencies were tested against each response. Monosynaptic IPSP(B)s were recorded in the presence of GABA(A), AMPA/kainate and NMDA receptor antagonists. All GABA(B) receptor antagonists tested depressed the IPSP(B) with an IC50 based rank order of potency of CGP55679> or =CGP56433 = CGP55845A = CGP52432>CGP51176>CGP36742. Paired-pulse EPSP widening was recorded as an index of paired-pulse depression of GABA-mediated IPSP/Cs. A similar rank order of potency of antagonism of paired-pulse widening was observed to that for IPSP(B) inhibition. Comparison of the IC50 values for IPSP(B) inhibition and paired-pulse EPSP widening revealed a close correlation between the two effects in that their IC50s lay within the 95% confidence limits of a correlation line that described IC50 values for inhibition of paired-pulse EPSP widening that were 7.3 times higher than those for IPSP(B) inhibition. Using the compounds tested here it is not possible to assign different subtypes of GABA(B) receptor to pre- and post-synaptic loci at GABAergic synapses. However, 5-10 fold higher concentrations of antagonist are required to block presynaptic as opposed to postsynaptic receptors when these are activated by synaptically released GABA. PMID- 10369476 TI - Mitogenic activation of human prostate-derived fibromuscular stromal cells by bradykinin. AB - Biologically active kinin peptides are released from precursor kininogens by kallikreins. Kinins act on kinin receptors to mediate diverse biological functions including smooth muscle contraction, inflammation, pain and mitogenicity. All components of the kallikrein-kinin system exist in human male genital secretions suggesting that these molecules participate in physiological and pathophysiological genitourinary function. The objective of this study was to assess the consequences of kinin action on prostate cells. Primary cultures of prostate secretory epithelial (PE) and prostate fibromuscular stromal (PS) cells were established from human prostate tissue. Transcripts encoding both the human B1 and B2 bradykinin receptor subtypes were detected in human prostate transition zone tissue and in cultured cells by RT-PCR. In receptor binding assays, the B1 subtype predominated on PE cell membranes and the B2 subtype predominated on PS cell membranes. In PS cells, but not in PE cells, BK induced significant inositol phosphate accumulation and [3H]-thymidine uptake. These responses were mediated through the B2 receptor subtype. The use of signal transduction inhibitors indicated that mitogenic activation by BK occurred through both protein kinase C (PKC) and protein tyrosine kinase dependent mechanisms. PMA (phorbol 12-myristate 13-acetate) produced maximal [3H]-thymidine uptake by PS cells, resulted in cell elongation and caused the alpha-actin fibres present in PS smooth muscle cells to became organized into parallel arrays along the length of the elongated cells. In summary, the prostate contains a functional kallikrein-kinin system, which could be significant in physiological and pathophysiological prostate function. PMID- 10369477 TI - Rate of change of blood concentrations is a major determinant of the pharmacodynamics of midazolam in rats. AB - The objective of this investigation was to characterize quantitatively the influence of the rate of increase in blood concentrations on the pharmacodynamics of midazolam in rats. The pharmacodynamics of midazolam were quantified by an integrated pharmacokinetic-pharmacodynamic modelling approach. Using a computer controlled infusion technique, a linear increase in blood concentrations up to 80 ng ml(-1) was obtained over different time intervals of 16 h, resulting in rates of rise of the blood concentrations of respectively, 1.25, 1.00, 0.87, 0.46, 0.34 and 0.20 ng ml(-1) min(-1). In one group of rats the midazolam concentration was immediately brought to 80 ng ml(-1) and maintained at that level for 4 h. Immediately after the pretreatment an intravenous bolus dose was given to determine the time course of the EEG effect in conjunction with the decline of midazolam concentrations. The increase in beta activity (11.5-30 Hz) of the EEG was used as pharmacodynamic endpoint. For each individual animal the relationship between blood concentration and the EEG effect could be described by the sigmoidal Emax model. After placebo, the values of the pharmacodynamic parameter estimates were Emax = 82+/-5 microV, EC50,u = 6.4+/-0.8 ng ml(-1) and Hill factor = 1.4+/-0.1. A bell-shaped relationship between the rate of change of midazolam concentration and the value of EC50,u was observed with a maximum of 21+/-5.0 ng ml(-1) at a rate of change of 0.46 ng ml(-1) min(-1); lower values of EC50,u were observed at both higher and lower rates. The findings of this study show that the rate of change in plasma concentrations is an important determinant of the pharmacodynamics of midazolam in rats. PMID- 10369478 TI - Pindolol-insensitive [3H]-5-hydroxytryptamine binding in the rat hypothalamus; identity with 5-hydroxytryptamine7 receptors. AB - Pindolol-insensitive [3H]-5-hydroxytryptamine ([3H]-5-HT) binding to rat hypothalamic membranes was pharmacologically and functionally characterized to resolve whether this procedure selectively labels 5-HT7 receptors. Consistent with a previous report, 3 microM and not 100 nM pindolol was required to occupy fully 5-HT1A and 5-HT1B receptors. Remaining [3H]-5-HT binding was saturable (KD, 1.59+/-0.21 nM; Bmax, 53.8+/-3.1 fmol x mg protein(-1)). Displacement of [3H]-5 HT with metergoline and 5-CT revealed shallow Hill slopes (<0.5) but seven other compounds had slopes >0.8 and pKi values and the rank order of affinity were significantly correlated (r = 0.81 and 0.93, respectively) with published [3H]-5 HT binding to rat recombinant 5-HT7 receptors. In the presence of pindolol, 5-HT enhanced accumulation of [32P]-cyclic AMP was unaffected by the 5-HT4 antagonist RS39604 (0.1 microM) or the 5-ht6 antagonist Ro 04-6790 (1 microM) but significantly attenuated by mesulergine (250 nM), ritanserin (450 nM) or methiothepin (200 nM) which have high affinity for the 5-HT7 receptor. Intracerebroventricular pretreatment with the serotonergic neurotoxin 5,7 dihydroxytryptamine, 5,7-DHT, elevated the [3H]-5-HT Bmax 2 fold, indicating that the hypothalamic 5-HT7 receptor is post-synaptic to 5-HT nerve terminals and regulated by synaptic 5-HT levels. These results suggest that, in the presence of 3 microM pindolol, [3H]-5-HT selectively labels hypothalamic binding sites consistent with functional 5-HT7 receptors. PMID- 10369479 TI - Inhibition of HERG channels stably expressed in a mammalian cell line by the antianginal agent perhexiline maleate. AB - Perhexiline has been used as an anti-anginal agent for over 25 years, and is known to cause QT prolongation and torsades de pointes. We hypothesized that the cellular basis for these effects was blockade of I(Kr). A stable transfection of HERG into a CHO-K1 cell line produced a delayed rectifier, potassium channel with similar properties to those reported for transient expression in Xenopus oocytes. Perhexiline caused voltage- and frequency-dependent block of HERG (IC50 7.8 microM). The rate of inactivation was increased and there was a 10 mV hyperpolarizing shift in the voltage-dependence of steady-state inactivation, suggestive of binding to the inactivated state. In conclusion, perhexiline potently inhibits transfected HERG channels and this is the probable mechanism for QT prolongation and torsades de pointes. Channel blockade shows greatest affinity for the inactivated state. PMID- 10369480 TI - In vitro alpha1-adrenoceptor pharmacology of Ro 70-0004 and RS-100329, novel alpha1A-adrenoceptor selective antagonists. AB - It has been hypothesized that in patients with benign prostatic hyperplasia, selective antagonism of the alpha1A-adrenoceptor-mediated contraction of lower urinary tract tissues may, via a selective relief of outlet obstruction, lead to an improvement in symptoms. The present study describes the alpha1-adrenoceptor (alpha1-AR) subtype selectivities of two novel alpha1-AR antagonists, Ro 70-0004 (aka RS-100975) and a structurally-related compound RS-100329, and compares them with those of prazosin and tamsulosin. Radioligand binding and second-messenger studies in intact CHO-K1 cells expressing human cloned alpha1A-, alpha1B- and alpha1D-AR showed nanomolar affinity and significant alpha1A-AR subtype selectivity for both Ro 70-0004 (pKi 8.9: 60 and 50 fold selectivity) and RS 100329 (pKi 9.6: 126 and 50 fold selectivity) over the alpha1B- and alpha1D-AR subtypes respectively. In contrast, prazosin and tamsulosin showed little subtype selectivity. Noradrenaline-induced contractions of human lower urinary tract (LUT) tissues or rabbit bladder neck were competitively antagonized by Ro 70-0004 (pA2 8.8 and 8.9), RS-100329 (pA2 9.2 and 9.2), tamsulosin (pA2 10.4 and 9.8) and prazosin (pA2 8.7 and 8.3 respectively). Affinity estimates for tamsulosin and prazosin in antagonizing alpha1-AR-mediated contractions of human renal artery (HRA) and rat aorta (RA) were similar to those observed in LUT tissues, whereas Ro 70-0004 and RS-100329 were approximately 100 fold less potent (pA2 values of 6.8/6.8 and 7.3/7.9 in HRA/RA respectively). The alpha1A-AR subtype selectivity of Ro 70-0004 and RS-100329, demonstrated in both cloned and native systems, should allow for an evaluation of the clinical utility of a 'uroselective' agent for the treatment of symptoms associated with benign prostatic hyperplasia. PMID- 10369481 TI - L-365,260 inhibits in vitro acid secretion by interacting with a PKA pathway. AB - The aim of this study was to analyse the antisecretory mechanism of L-365,260 in vitro in isolated rabbit gastric glands. We showed that compound L-365,260, described as a non-peptide specific competitive CCK-B receptor antagonist, was able to dose-dependently inhibit [14C]-aminopyrine accumulation induced by histamine (10(-4) M), carbachol (5x10(-5) M), 3-isobutyl-1-methyl-xanthine (IBMX) (5x10(-6) M) and forskolin (5x10(-7) M) with similar IC50 values respectively of 1.1+/-0.6x10(-7) M, 1.9+/-1.2x10(-7) M, 4.2+/-2.0x10(-7) M and 4.0+/-2.8x10(-7) M. We showed that L-365,260 acted beyond receptor activation and production of intracellular second messengers and that it had no action on the H+/K+ -ATPase. We found that L-365,260 inhibited cyclic AMP-induced [14C]-aminopyrine accumulation in digitonin-permeabilized rabbit gastric glands, suggesting that this compound acted, at least in part, as an inhibitor of the cyclic AMP dependent protein kinase (PKA) pathway. PMID- 10369482 TI - Characterization of the decrease of extracellular striatal dopamine induced by intrastriatal morphine administration. AB - The effect of intrastriatally-administered morphine on striatal dopamine (DA) release was studied in freely moving rats. Morphine (1, 10 or 100 microM) was given into the striatum by reversed microdialysis, and concentrations of DA and its metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) were simultaneously measured from the striatal dialysates. Intrastriatally administered morphine significantly and dose-dependently decreased the extracellular concentration of DA, the concentrations of the acidic DA metabolites were only slightly decreased. The effect of morphine was antagonized by naltrexone (2.25 mg kg(-1), s.c.). Pretreatment with a preferential kappa opioid receptor antagonist, MR2266 [(-)-5,9 alpha-diethyl-2-(3-furylmethyl)-2' hydroxy-6,7-benzomorphane; 1 mg kg(-1), s.c.], had no effect on the decrease of extracellular DA evoked by intrastriatal morphine (100 microM). Intrastriatal administration of the selective micro-opioid receptor agonist [D-Ala2,MePhe4,Gly ol5] enkephalin (DAMGO; 1 microM), significantly decreased the extracellular concentration of DA in the striatum. When the rats were given morphine repeatedly in increasing doses (10-25 mg kg(-1), s.c.) twice daily for 7 days and withdrawn for 48 h, the decrease of extracellular DA induced by morphine (100 microM) was significantly less than that seen in saline-treated controls. Our results show that besides the well-known stimulatory effect there is a local inhibitory component in the action of morphine on striatal DA release in the terminal regions of nigrostriatal DA neurones. Tolerance develops to this inhibitory effect during repeated morphine treatment. Furthermore, our results suggest that the effect of intrastriatally-administered morphine is mediated by the micro opioid receptors. PMID- 10369483 TI - Control of glutamate release by calcium channels and kappa-opioid receptors in rodent and primate striatum. AB - The modulation of depolarization (4-aminopyridine, 2 mM)-evoked endogenous glutamate release by kappa-opioid receptor activation and blockade of voltage dependent Ca2+ -channels has been investigated in synaptosomes prepared from rat and marmoset striatum. 4-Aminopyridine (4-AP)-stimulated, Ca2+ -dependent glutamate release was inhibited by enadoline, a selective kappa-opioid receptor agonist, in a concentration-dependent and norbinaltorphimine (nor-BNI, selective kappa-opioid receptor antagonist)-sensitive manner in rat (IC50 = 4.4+/-0.4 microM) and marmoset (IC50 = 2.9+/-0.7 microM) striatal synaptosomes. However, in the marmoset, there was a significant (approximately 23%) nor-BNI-insensitive component. In rat striatal synaptosomes, the Ca2+ -channel antagonists omega agatoxin-IVA (P/Q-type blocker), omega-conotoxin-MVIIC (N/P/Q-type blocker) and omega-conotoxin-GVIA (N-type blocker) reduced 4-AP-stimulated, Ca2+ -dependent glutamate release in a concentration-dependent manner with IC50 values of 6.5+/ 0.9 nM, 75.5+5.9 nM and 106.5+/-8.7 nM, respectively. In marmoset striatal synaptosomes, 4-AP-stimulated, Ca2+ -dependent glutamate release was significantly inhibited by omega-agatoxin-IVA (30 nM, 57.6+/-2.3%, inhibition), omega-conotoxin-MVIIC (300 nM, 57.8+/-3.1%) and omega-conotoxin-GVIA (1 microM, 56.7+/-2%). Studies utilizing combinations of Ca2+ -channel antagonists suggests that in the rat striatum, two relatively distinct pools of glutamate, released by activation of either P or Q-type Ca2+ -channels, exist. In contrast, in the primate there is much overlap between the glutamate released by P and Q-type Ca2+ -channel activation. Studies using combinations of enadoline and the Ca2+ channel antagonists suggest that enadoline-induced inhibition of glutamate release occurs primarily via reduction of Ca2+ -influx through P-type Ca2+ channels in the rat but via N-type Ca2+ -channels in the marmoset. In conclusion, the results presented suggest that there are species differences in the control of glutamate release by kappa-opioid receptors and Ca2+ -channels. PMID- 10369484 TI - A study of NPY-mediated contractions of the porcine isolated ear artery. AB - Enhanced contractions of the porcine isolated ear artery by the alpha2 adrenoceptor agonist UK14304 are uncovered by pharmacological manipulation. As both neuropeptide Y (NPY) receptors and alpha2-adrenoceptors are negatively coupled to adenylyl cyclase in this tissue, we determined whether NPY is also able to produce an enhanced contraction in the same tissue, under the same conditions. NPY (0.1 microM) produced a small contraction of porcine isolated ear arteries which was 5.1+/-0.8% of the response to 60 mM KCl (n = 14). An enhanced NPY response was uncovered if the tissue was pre-contracted with 0.1 microM U46619, and relaxed back to baseline with 1-2 microM forskolin before the addition of NPY (49.8+/-5.3%, n = 14). Forskolin (1 microM) stimulated cyclic AMP accumulation in porcine ear artery segments in the presence of 0.1 microM U46619 and 1 mM isobutylmethylxanthine (IBMX), NPY (0.1 microM) inhibited this response by 40%, but had no effect on basal levels of cyclic AMP. An enhanced response to 0.1 microM NPY was also obtained after pre-contraction with 0.1 microM U46619 and relaxation with either SNP (28.9+/-5.7%, n = 14), or dibutyryl cyclic AMP (21.2+/ 4.6%, n = 14). This indicates that at least part of the enhanced response to NPY is independent of the agonist's ability to inhibit adenylyl cyclase. In conclusion, an enhanced contraction to NPY in the porcine isolated ear artery can be obtained by prior pharmacological manipulation. The enhanced responses are mediated through adenylyl cyclase-dependent and independent pathways similar to those reported for alpha2-adrenoceptors in this preparation. PMID- 10369485 TI - Potentiation of cyclic AMP-mediated vasorelaxation by phenylephrine in pulmonary arteries of the rat. AB - Alpha1-adrenoceptor agonists may potentiate relaxation to beta-adrenoceptor agonists, although the mechanisms are unclear. We compared relaxations induced by beta-adrenoceptor agonists and cyclic AMP-dependent vasodilators in rat pulmonary arteries constricted with prostaglandin F2alpha (PGF2alpha) or the alpha1 adrenoceptor agonist phenylephrine (PE). In addition, we examined whether differences were related to cyclic AMP- or nitric oxide (NO) and cyclic GMP dependent pathways. Isoprenaline-induced relaxation was substantially potentiated in arteries constricted with PE compared with PGF2alpha. Methoxamine was similar to PE, whereas there was no difference between PGF2alpha and 30 mM KCl. The potentiation was primarily due to a marked increase in the NO-independent component of relaxation, from 9.1+/-1.7% for PGF2alpha to 55.1+/-4.4% for PE. NO dependent relaxation was also enhanced, but to a lesser extent (50%). Relaxation to salbutamol was almost entirely NO-dependent in both groups, and was potentiated approximately 50% by PE. Relaxation to forskolin (activator of adenylate cyclase) was also enhanced in PE constricted arteries. Part of this relaxation was NO-dependent, but the major effect of PE was to increase the NO independent component. Propranolol diminished but did not abolish the potentiation. There was no difference in response to CPT cyclic AMP (membrane permeant analogue) between PE and PGF2alpha, suggesting that mechanisms distal to the production of cyclic AMP were unchanged. Relaxation to sodium nitroprusside (SNP) was the same for PE and PGF2alpha, although relaxation to acetylcholine (ACh) was slightly depressed. This implies that potentiation by PE does not involve the cyclic GMP pathway directly. Mesenteric arteries constricted with PE did not show potentiation of isoprenaline-induced relaxation compared to those constricted with PGF2alpha, suggesting that this effect may be specific to the pulmonary circulation. These results clearly show that PE potentiates both the NO independent and -dependent components of cyclic AMP-mediated relaxation in pulmonary arteries of the rat, although the effect on the former is more profound. We suggest that potentiation of both components is largely due to direct activation of adenylate cyclase via alpha1-adrenoceptors, within the smooth muscle and endothelial cells respectively. PMID- 10369486 TI - Further characterization of the ORL1 receptor-mediated inhibition of noradrenaline release in the mouse brain in vitro. AB - Mouse brain slices preincubated with [3H]-noradrenaline or [3H]-serotonin were superfused with medium containing naloxone 10 microM; we studied whether nociceptin (the endogenous ligand at ORL1 receptors) affects monoamine release. Furthermore, the affinities of ORL1 ligands were determined using [3H]-nociceptin binding. The electrically (0.3 Hz) evoked tritium overflow in mouse cortex slices preincubated with [3H]-noradrenaline was inhibited by nociceptin and [Tyr14] nociceptin (maximally by 80%; pEC50 7.52 and 8.28) but not affected by [des-Phe1] nociceptin (pEC50<6). The ORL1 antagonist naloxone benzoylhydrazone antagonized the effect of nociceptin and [Tyr14]-nociceptin. The effect of nociceptin did not desensitize, was not affected by blockade of NO synthase, cyclooxygenase and P1 purinoceptors and was decreased by the alpha2-adrenoceptor agonist talipexole. Nociceptin also inhibited the evoked overflow in mouse cerebellar, hippocampal and hypothalamic slices in a manner sensitive to naloxone benzoylhydrazone. The electrically (3 Hz) evoked tritium overflow in mouse cortex slices preincubated with [3H]-serotonin was inhibited by nociceptin; naloxone benzoylhydrazone antagonized this effect. The affinities (pKi) for [3H]-nociceptin binding to mouse cortex membranes were: nociceptin, 8.71; [Tyr14]-nociceptin, 9.82; [des Phe1]-nociceptin, <5.5; naloxone benzoylhydrazone, 5.85; naloxone, <4.5. In conclusion, nociceptin inhibits noradrenaline release in the mouse cortex via ORL1 receptors, which interact with presynaptic alpha2-autoreceptors on noradrenergic neurones. The effect of nociceptin does not desensitize nor does it involve NO, prostanoids or adenosine. Nociceptin also attenuates noradrenaline release from several subcortical regions and serotonin release from cortical slices by a naloxone benzoylhydrazone-sensitive mechanism. PMID- 10369488 TI - Lactic acid bacteria and the human gastrointestinal tract. AB - OBJECTIVE: This review summarises the effects of lactic acid bacteria on lactose malabsorption, bacterial/viral or antibiotic associated diarrhoea, and describes the impact of lactic acid bacteria on cancer and the fermentative products in the colon. RESULTS: Eight studies (including 78 patients) demonstrated that lactase deficient subjects absorbed lactose in yogurt better than lactose in milk, while two studies (25 patients) did not support this. Two studies (22 patients) showed that unfermented acidophilus milk was absorbed better than milk, while six studies (68 patients) found no significant differences. Addition of lactose hydrolysing enzyme, lactase, to milk improved lactose malabsorption in seven studies (131 lactose malabsorbers), while one study (10 malabsorbers) demonstrated no improvement. Lactic acid bacteria alleviated travellers' diarrhoea in one study (94 individuals) while a study including 756 individuals was borderline statistically significant. One study (50 individuals) did not find an effect of lactic acid bacteria on travellers' diarrhoea. Six studies (404 infants) demonstrated a significant effect of lactic acid bacteria on infant diarrhoea, while one study (40 infants) did not. Lactic acid bacteria moderated antibiotic associated diarrhoea in three studies (66 individuals), while two studies (117 individuals) were insignificant. CONCLUSIONS: Lactase deficient subjects benefit from a better lactose absorption after ingestion of yoghurt compared with milk and from milk added lactase, whereas ingestion of unfermented acidophilus milk does not seem to improve lactose absorption. The majority of studies support that lactic acid bacteria alleviate bacterial/viral induced diarrhoea, especially in infants, while the effect on antibiotic associated diarrhoea is less clear. Experimental studies indicate an effect of lactic bacteria on human cell cancer lines, but clinical evidence is lacking. A 'stabilising' effect of lactic acid bacteria on the colonic flora has not been documented. PMID- 10369489 TI - Preferences, quantities and concerns: socio-cultural perspectives on the gendered consumption of foods. AB - A review of the sociological research regarding the gendered features of food consumption is presented. The focus is upon differences between women and men in regard to their preferences for particular foods and types of meals, seen in relation to the cultural function of foods as symbolic markers of femininity or masculinity, assessments of the quantities of food consumed by women and men respectively, and differences between women and men in regard to concerns with food safety, health, weight reduction and fitness. Some methodological limitations of this research are discussed with particular reference to the need for interdisciplinary cooperation between sociologists and nutritionists in the design and analysis of dietary surveys. Suggestions are made in regard to future directions for sociological research in this field, with particular reference to the issue that dietary recommendations appear to focus upon increasing the consumption of foods that are markers of femininity and decreasing the consumption of foods that are markers of masculinity in Western food culture. PMID- 10369487 TI - Nitric oxide-dependent relaxation induced by M1 muscarinic receptor activation in the rat small intestine. AB - The aim of the present study was to investigate whether muscarinic M1 receptor activation induces intestinal relaxation via nerve-dependent nitric oxide formation. Mechanical activity in longitudinal segments of rat jejunum was recorded isotonically in organ baths. The muscarinic M1 receptor agonist 4-[[[(3 Chlorophenyl)amino]carbonyl]oxy]-N,N,N,-trimethyl-2-butyn- 1-ammonium chloride (McN-A-343, 10(-7)-10(-4) M) induced a concentration-dependent relaxation of rat jejunum. Relaxations induced by McN-A-343 (10(-5) M) were inhibited by the M1 receptor antagonist telenzepine (10(-8) M), and enhanced by the M3 receptor antagonist para-fluorohexahydrosiladifenidol (p-F-HHSiD; 3x10(-7) M). The inhibitory responses induced by McN-A-343 were abolished by the nitric oxide synthase inhibitors Nomega-nitro-L-arginine (L-NOARG; 10(-4) M) and Nomega monomethyl-L-arginine (L-NMMA; 3x10(-5) M), the guanylyl cyclase inhibitor 1H [1,2,4]oxadiazolo[4,3,-a]quinoxalin-1-one (ODQ; 10(-5) M), and by tetrodotoxin (TTX; 3x10(-7) M). Guanethidine or hexamethonium did not affect inhibitory responses induced by McN-A-343. In conclusion, McN-A-343 induces nerve-dependent, nitrergic relaxations in rat jejunum, via activation of muscarinic M1 receptors. Hence, selective muscarinic M1 receptor agonists or antagonists might offer possibilities for pharmacological manipulation of the NO system. PMID- 10369490 TI - Bioavailability of starch in bread rich in amylose: metabolic responses in healthy subjects and starch structure. AB - OBJECTIVE: This study investigated whether postprandial metabolic responses to bread could be lowered by substituting high amylose maize starch for a part of the flour. DESIGN AND SUBJECTS: Eight healthy subjects consumed test meals of equivalent nutritional composition based on white wheat bread, bread rich in amylose (HAWB) and spaghetti as a breakfast meal. Blood samples were collected to measure insulin and glucose concentration during two hours after consumption. The degree of starch crystallinity was investigated by X-ray diffraction and DSC analysis. RESULTS: HAWB produced low glycaemic (60 +/- 18) and insulinaemic (57 +/- 20) indexes similar to those of spaghetti (83 +/- 46, 61 +/- 16). In vitro amylase hydrolysis of the three foods showed that high amylose content in HAWB significantly lowered starch degradation in bread without affecting hydrolysis kinetics. Addition of amylose in dough increased the resistant starch content of HAWB (14% of dry matter). The resistant starch fraction was mainly composed of crystalline amylose (B-type X-ray diffraction pattern, melting temperature 105 degrees C) attributable to native high amylose maize starch incompletely gelatinised during bread-cooking. CONCLUSIONS: Bread produced by the substitution of high amylose maize starch for a part of wheat flour showed a low glycaemic index. Resistant starch in HAWB corresponded to native crystalline amylose not gelatinised during normal bread-processing conditions. PMID- 10369492 TI - Does degree of obesity influence the validity of reported energy and protein intake? Results from the SOS Dietary Questionnaire. Swedish Obese Subjects. AB - OBJECTIVE: To test the validity of a dietary questionnaire which was developed with the particular goal of measuring dietary intake in obese subjects. DESIGN: Reported energy intake was compared with 24 h energy expenditure measured in a chamber for indirect calorimetry (24 EE) and reported nitrogen intake with nitrogen in urine collected during the 24 h in the chamber. SUBJECTS: Twenty-nine overweight men and women, body mass index (BMI) ranging from 25.5 49.5 kg/m2. RESULTS: Reported energy intake correlated significantly with 24 EE (r = 0.50, P = 0.006) and reported urinary nitrogen correlated significantly with urinary nitrogen excretion (r=0.56, P=0.0015). Mean reported energy intake+/-s.d. was 10.2+/-3.6 MJ and mean 24 EEi s.d. was 10.3+/-1.9 MJ. Although this difference was small and non significant, it indicates some underreporting if one can assume that these overweight subjects are less physically active in the chamber than in free-living conditions. Reported nitrogen intake also suggested underreporting at the group level. However, when the data were analysed at the individual level it was clear that the underreporting errors did not increase with increasing degree of obesity. CONCLUSIONS: Previous studies with the SOS dietary questionnaire have demonstrated that it is possible to obtain plausible energy intakes from both obese and nonobese subjects. This present analysis further demonstrates that the questionnaire discriminates overweight subjects with high and low intakes of energy and protein, using unbiased biomarkers to judge validity. These data provide additional support for the usefulness of the SOS dietary questionnaire. PMID- 10369491 TI - Do iron and vitamin C co-supplementation influence platelet function or LDL oxidizability in healthy volunteers? AB - OBJECTIVE: To examine the effect of co-supplementation with iron and vitamin C on antioxidant status, platelet function and low density lipoprotein oxidation in normal healthy volunteers. DESIGN: The study was carried out with two groups of 20 subjects each acting as their own control, comparing presupplemention with postsupplemention. One group was supplemented with iron and the RDA level of vitamin C and the second group with iron and 260 mg/d vitamin C. SETTING: The International Antioxidant Research Centre, The Guy's, King's College and St Thomas's School of Biomedical Science, Guy's Campus, London. SUBJECTS: Forty normal healthy volunteers, recruited from the staff of the Medical School and Hospital in which two volunteers withdrew during the study. INTERVENTIONS: Subjects in both studies were randomly assigned to one of two groups (5 males and 5 females group) and received supplements containing iron (14 mg/d) and either 60 mg/d (Group A) or 260 mg/d (Group B) vitamin C for 12 wk. Blood samples were taken at 6 wk and 12 wk, and prior to supplementation and analysed for iron and antioxidant status (transferrin bound iron, vitamin C and E, and beta-carotene levels) in both studies. Samples from the first study were analysed for the susceptibility of LDL isolated from plasma to Cu2+-induced oxidation and samples from the second for platelet function. RESULTS: Transferrin-bound iron was significantly increased (P < 0.05) at 12 wk, in Group A subjects (from 14.9+/-5.3 micromol/1 to 19.5+/-2.3 micromol/l; mean+/-s.d.; n=19), whereas those in Group B showed a significant increase (P < 0.05) after 6 wk (from 15.8+/-4.5 micromol/l to 20.4+/-6.6 micromol/l; n = 19) which decreased at 12 wk (16.3+/-5.0 micromol/l). Plasma total ascorbate significantly increased from an initial level of 59.3+/-21.3 micromol/l to 87.6+/-29.0 micromol/l after 6 wk and 81.7+/-11.4 micromol/l after 12 wk following the Group B supplementation, but only after 12 wk in Group A (from 64.0+/-24.8 micromol/l to 77.2+/-13.2 micromol/l). Plasma alpha-tocopherol concentrations were significantly increased after 6 wk and 12 wk with both levels of supplementation (from 24.2+/-5.71 micromol/l Group A and 23.4+/-5.3 micromol/l Group B to 26.3+/-5.5 micromol/l and 25.71+/-4.7 micromol/1 respectively at 12wk). The mean lag phase to oxidation of low density lipoprotein (LDL) was significantly increased in subjects in Group B after 12 wk ingestion of iron and 260 mg vitamin C (from 80.0+/-14.8 min to 97.2+/-16.9 min; n = 9). Platelet sensitivity to ADP-induced aggregation was significantly decreased (P < 0.05) by 12 wk in Group A (from EC50 2.3 < or = 1.3 microM to 3.7+/-2.2 microM; n = 10), whereas those receiving higher vitamin C showed a significant decrease (P < 0.05; from EC50 1.9+/-0.6 microM to 3.1+/-1.8 microM) after 6wk which subsequently increased towards presupplemental levels (2.6+/-1.6 microM). Platelets from the latter subjects showed a significant reduction in ADP-induced ATP secretion at both 6wk and 12 wk. CONCLUSION: The results show modest beneficial effects on LDL oxidation and platelet function following supplementation with iron and vitamin C. No evidence for pro-oxidant effects was observed. PMID- 10369493 TI - Randomized, double-blind trial of chitosan for body weight reduction. AB - BACKGROUND: Overweight and obesity is a prevalent and costly threat to public health. Compelling evidence links overweight and obesity with serious disorders such as cardiovascular diseases and diabetes. Dietary regimen are notoriously burdened with poor compliance. Chitosan is promoted in the US and other countries as an oral remedy to reduce fat absorption and has now been incorporated as a major constituent into several over-the-counter remedies. The primary aim of this study is to investigate the clinical effectiveness of oral chitosan for body weight reduction. METHODS: Thirty-four overweight volunteers were included in a randomized placebo-controlled double-blind trial. Subjects were assigned to receive either four capsules of chitosan or indistinguishable placebo twice daily for 28 consecutive days. Measurements were taken at baseline, after 14 and 28d of treatment. Subjects maintained their normal diet and documented the type and amount of food consumed. Adverse effects were assessed and compliance monitored. RESULTS: Data from 30 subjects were entered into an intention-to-treat analysis. After four weeks of treatment, body mass index, serum cholesterol, triglycerides, vitamin A, D, E and beta-carotene were not significantly different in subjects receiving chitosan compared to those receiving placebo. Vitamin K was significantly increased after four weeks in the chitosan group compared with placebo (P<0.05). Compliance was 91.5% and 96.0% for chitosan and placebo groups respectively. CONCLUSION: The above data suggest that chitosan in the administered dosage, without dietary alterations, does not reduce body weight in overweight subjects. No serious adverse effects were reported. PMID- 10369494 TI - Body composition in children and adults by air displacement plethysmography. AB - OBJECTIVES: Air displacement plethysmography (ADP) may provide a partial alternative to body density (Bd) and therefore body composition measurement compared to conventional hydrodensitometry (Hd) in children. As there are no evaluation studies of ADP in children, this study had a two-fold objective: to compare Bd estimates by ADP and Hd; and to compare fat estimates by both ADP and Hd to fat estimates by another reference method, dual energy X-ray absorptiometry (DXA). SETTING: Obesity Research Center, St. Luke's/Roosevelt Hospital, New York, USA. SUBJECTS: One hundred and twenty subjects (66 females/54 males) who ranged in age from 6-86 y and in body mass index (BMI, kg/m2) from 14.1-40.0 kg/m2 met study entry criteria. STUDY DESIGN: Cross-sectional study of healthy children (age < or = 19 y) and adult group for comparison to earlier studies. Each subject completed ADP, Hd, and DXA studies on the same day. Only subjects with subjectively-judged successful Hd studies were entered into the study cohort. RESULTS: There was a high correlation between Bd by ADP and Hd (Bd Hd = 0.11 + 0.896 x Bd ADP; r = 0.93, SEE = 0.008 g/cm3, P < 0.0001), although the regression line slope and intercept differed significantly from 1 and 0, respectively. Additional analyses localized a small-magnitude Bd bias in the child (n = 48) subgroup. Both ADP and Hd %fat estimates were highly correlated (r > 0.9, P < 0.0001) with %fat by DXA in child and adult subgroups. Bland-Altman analyses revealed no significant %fat bias by either ADP or Hd vs DXA in either children or adults, although a bias trend (P = 0.11) was detected in the child subgroup. CONCLUSION: With additional refinements, the air displacement plethysmography system has the potential of providing an accurate and practical method of quantifying body fat in children as it now does in adults. PMID- 10369495 TI - Determinants of salt use in cooked meals in The Netherlands: attitudes and practices of food preparers. AB - OBJECTIVE: To assess current habits of using salt and other seasonings in food preparation, and to investigate the psychosocial determinants of salt use. In addition we examined differences in salt consumption and psychosocial determinants with respect to stages of change and socio-economic classes. SUBJECTS AND METHODS: 400 adults were interviewed to determine: 1) use of salt and other seasonings in food preparation; 2) added table salt; 3) consumption of foods with high salt content; and 4) psychosocial determinants of salt consumption. Subjects were divided into 5 stages of change for salt content of the cooked meal: precontemplation, contemplation, decision, action and maintenance. RESULTS: Salt appeared to be the type of seasoning mostly added in food preparation. Frequently used seasonings containing no added salt were pepper, onion, nutmeg, garlic, curry, sweet pepper powder, parsley, and bay-leaf. Spearman correlation coefficients between added cooking salt and taste attitude, health attitude, social influence and self-efficacy were 0.51, 0.14, 0.36, and 0.32 respectively. Correlations between consumption of foods with high salt content and convenience attitude, taste attitude, health attitude and self efficacy were 0.41, 0.27, 0.18 and 0.21 respectively. Subjects in maintenance of low salt consumption, used less salt than subjects in precontemplation. Socio economic status only showed an effect on the use of foods with high salt content. CONCLUSION AND PRACTICAL IMPLICATIONS: Health aspects did not play a major role in salt intake, whereas taste attitude was an important predictor of added cooking salt. Attempts to reduce cooking salt should be directed at satisfying taste. Nutrition education tailored to stages of change may be very helpful in decreasing salt consumption. PMID- 10369496 TI - Changes in dietary intake account for seasonal changes in cardiovascular disease risk factors. AB - OBJECTIVES: (1) to compare dietary intake in summer and winter time; (2) to measure the change in body mass index (BMI), blood pressure and serum cholesterol between winter and summer; and (3) to determine the relationships between seasonal differences in dietary intake and BMI, blood pressure and serum cholesterol measurements. SUBJECTS AND METHODS: Ninety-four male industrial employees were screened twice in one year, in their work place, at winter and summer time. Workers were recruited from two factories and response rate was 95%. Health-related variables, including dietary intake, blood pressure and serum cholesterol were evaluated at each season and were compared. Correlation coefficients between seasonal differences in dietary intake and in BMI, blood pressure and serum cholesterol were calculated. RESULTS: From summer to winter the mean values of BMI increase from 26.1 kg/cm2 to 26.6 (P=0.038), systolic blood pressure from 119.6 to 121.6 (P=0.025), diastolic blood pressure from 75.2 to 77.2 mmHg (P=0.001), total cholesterol from 200.8 to 208.6 mg/dL (P=0.001), LDL cholesterol from 125.2 to 134.9 (P=0.001) and HDL cholesterol from 42.7 to 44.3 (P=0.0084). Triglycerides levels decrease from 174 to 145 in the winter (P=0.03). Mean dietary intake of fat increases from 99.1 to 106.0 (P=0.0016), saturated fat from 43.6 to 46.3 (P=0.0137), polyunsaturated fat from 25.1 to 28.3 (P=0.0002), cholesterol from 462.0 to 497.9 (P=0.0313), sodium from 5778.5 to 8208.2 (P=0.0035), zinc from 11.6 to 12.3 (P=0.0001), vitamin B1 from 1.4 to 1.5 (P=0.002), vitamin D from 4.3 to 4.9 (P=0.0323) and vitamin E from 11.2 to 12.7 (P=0.0073). Significant correlation was shown between the seasonal increase in saturated fat and the increase in BMI (r=0.37), total cholesterol (r=0.21) and LDL cholesterol (r=0.29). Seasonal change in dietary cholesterol intake was significantly and positively correlated with serum total cholesterol (r=0.24) and LDL cholesterol (r=0.24). Blood pressure was not associated with nutritional intake variables. CONCLUSIONS: Dietary intake in summer and winter is different as well as blood pressure, BMI and serum cholesterol. The seasonal increase in fat and cholesterol intake at winter time is associated with changes in BMI and serum cholesterol. PMID- 10369497 TI - Day-to-day and within-day variation in urinary iodine excretion. AB - OBJECTIVE: To examine the day-to-day and within-day variation in urinary iodine excretion and the day-to-day variation in iodine intake. DESIGN: Collection of consecutive 24-h urine samples and casual urine samples over 24h. SETTING: The study population consisted of highly motivated subjects from our Institute. SUBJECTS: Study 1: Ten healthy subjects (seven females and three males) aged 30 46 y. Study 2: Twenty-two healthy subjects (9 males and 13 females) aged 30-55 y. METHODS: Study 1: 24-h urine samples were collected for four consecutive days. Study 2: Each urine voided over 24 h was collected into separate containers. In both studies dietary records were kept. MAIN OUTCOME MEASURES: Twenty-four-hour urinary iodine excretion, 24-h urinary iodine excretion estimated as I/Cr*24 h Cr and as a concentration in casual urine samples. RESULTS: Study 1: Both iodine excreted in 24-h urine and iodine intake varied from day-to-day. Iodine excretion correlated with iodine intake (=-0.46, P=0.01). Iodine intake (mean 89 +/- 6.5 microg/d) was not significantly different from iodine excretion (mean 95 +/- 5.3 microg/d). Study 2: Twenty-four hour iodine excretion estimated as I/Cr*24 h Cr from the morning urine sample was significantly lower than actual 24-h iodine excretion, whereas 24-h iodine excretion estimated as I/Cr*24 h Cr from the first sample after the morning sample and the last sample before the subjects went to bed was not significantly different from actual 24-h iodine excretion. Twenty four-hour urine excretion estimated as a concentration was lower than actual 24-h iodine excretion in casual urine taken at any time of the day. CONCLUSIONS: For determination of iodine status in an individual, more than one 24-h urine sample must be used. The use of the I/Cr ratio in casual urine samples is a usable measure of iodine status if corrected for the age- and sex-adjusted 24-h creatinine excretion. Further, the study suggests that fasting morning urine samples would underestimate iodine status in this population. PMID- 10369498 TI - Effect of dietary copper intakes on biochemical markers of bone metabolism in healthy adult males. AB - OBJECTIVE: To investigate the effects of changing from a medium (1.6 mg Cu/d) to a low (0.7 mg Cu/d) or a high (6.0 mg/d) Cu intake on biochemical indices of bone turnover in healthy adult males. DESIGN: A longitudinal intervention trial. SETTING: The study was conducted at the Institute of Food Research, Norwich, UK. SUBJECTS: Eleven healthy adult males aged 20-59 y were recruited from Norwich Research Park. INTERVENTION: Subjects were given medium (1.6 mg/d), low (0.7 mg/d) and high (6.0 mg/d) intakes of Cu, in that order, over consecutive 8 week periods with a minimum of 4 week washout periods. On the last d of each dietary period fasting first void urine and blood were collected. RESULTS: Serum caeruloplasmin or Cu (indices of Cu status), serum osteocalcin (biomarker of bone formation), urinary creatinine (Cr) were unaffected by dietary Cu intake. Urinary Pyr/Cr and Dpyr/Cr (biomarkers of bone resorption) were significantly increased (P < 0.05) (by 30% and 25%, respectively), when subjects were switched from the medium to the low Cu diet and were significantly decreased (P < 0.05) (by 30%) and 22% respectively), when subjects were switched from the low to the high Cu diet. CONCLUSION: The findings of the present study could have implications for bone health in individuals with marginal Cu intakes. Thus, further studies are warranted to better define the relationship of marginal Cu intakes to bone health. PMID- 10369499 TI - Dietary counselling effectively improves lipid levels in patients with endogenous hypertriglyceridemia: emphasis on weight reduction and alcohol limitation. AB - OBJECTIVE: To evaluate the short-term effect of dietary counselling in patients with endogenous hypertriglyceridemia and evaluate the effects of advised nutrient changes. DESIGN: A prospective dietary intervention study in patients with endogenous hypertriglyceridemia from January I st 1988 to December 31 st 1996 according to the Dutch guidelines for a healthy diet. Before and after the dietary intervention period of 12 weeks, 24h food recalls were used to assess dietary intake and macronutrient composition. Effectiveness was evaluated by assessment of body weight, serum lipids, lipoproteins and insulin resistance parameters. SETTING: Leiden outpatient Lipid Clinic. SUBJECTS: Forty-five newly diagnosed, untreated patients with endogenous hypertriglyceridemia. RESULTS: A significant reduction in energy intake and body weight as well as changes in macronutrient composition were observed. Total serum triacylglycerol and cholesterol levels decreased by 31% and 15%, respectively. No effects were observed on serum glucose and insulin levels. Weight reduction was significantly correlated with reduction of total plasma triacylglycerol levels and inversely correlated with changes in HDL cholesterol levels. Of all nutrients assessed, only reduction of alcohol intake correlated with improvement of total serum triacylglycerol. CONCLUSIONS: Short-term dietary counselling in patients with endogenous hypertriglyceridemia can effectively improve serum lipid and lipoprotein levels. With regard to the advised nutrient changes, weight loss and limitation of alcohol intake prove to be the best predictors of triacylglycerol reduction. PMID- 10369500 TI - Models of antioxidant protection against biological oxidative damage. PMID- 10369501 TI - Presenting statistical uncertainty in trends and dose-response relations. AB - When one estimates the effects of a polytomous exposure, it is common practice to express all effects relative to a baseline or reference level. Certain authors have challenged this practice and proposed alternatives, which we review here. One alternative, the "floating absolute risk" method, can supply useful statistics and trend graphs, but it does not yield valid confidence intervals for relative risks. All categorical methods have further shortcomings when the exposure is continuous, however. These shortcomings can be addressed by plotting or tabulating confidence limits for points on a flexible curve fitted to the uncategorized data. PMID- 10369502 TI - Look before you leap: stratify before you standardize. AB - This paper presents a mathematical model to show the conditions in which age standardization can be used to summarize age-specific rates for comparison purposes over calendar time. It shows that the conditions for valid comparison depend on the type of measure used for comparison, that is, difference, ratio, or percent change. If the measure for comparison is a difference of the standardized rates at two time points, then the age-specific rates need to maintain a constant rate difference over time for the comparison to be valid. If the measure for comparison is a ratio or percent change of the standardized rates at two time points, then the age-specific rates need to maintain a constant rate ratio over time for the comparison to be valid. Since in reality, as shown by our Canadian empirical data, age-specific rates do not always maintain a consistent pattern over time, it is recommended that one should always stratify the data to look at patterns of age-specific rates before applying age standardization. PMID- 10369503 TI - Black-white differences in disability and morbidity in the last years of life. AB - To assess black-white differences in disability and morbidity in the last years of life, the authors analyzed data from the National Health Interview Survey from 1986 to 1994, with mortality follow-up through December 1995. A baseline household interview was conducted for 10,187 decedents aged 50 years and over within 2 years before death. Data collected included long-term limitation of activity, number of chronic conditions, number of bed days, doctor visits, and days of short hospital stay during the year preceding the interview. For both blacks and whites, educational attainment was inversely associated with disability/morbidity indices. Black decedents had greater morbidity compared with whites, and this difference was consistent across educational levels. Adjustment for education reduced the black-white difference in limitation of activity score by 32%, bed days by 59%, and hospital stay days by 40%. This study from a national representative US sample indicates that black decedents experienced greater disability/morbidity and worse quality of life through their last few months or years of life. Educational attainment was associated with morbidity before death and accounted for much of the black-white difference. PMID- 10369504 TI - The Women's Health Trial Feasibility Study in Minority Populations: changes in dietary intakes. AB - This randomized clinical trial examined the feasibility of low-fat dietary interventions among postmenopausal women of diverse backgrounds. During 1992 1994, 2,208 women aged 50-79 years, 28% of whom were black and 16% Hispanic, enrolled at clinics in Atlanta, Georgia, Birmingham, Alabama, and Miami, Florida. Intervention/support groups met periodically with a nutritionist to reduce fat intake to 20% of energy and to make other diet modifications. At 6 months postrandomization, the intervention group reduced fat intake from 39.7% of energy at baseline to 26.4%, a reduction of 13.3% of energy, compared with 2.3% among controls. Saturated fatty acid and cholesterol intakes were reduced, but intakes of fruits and vegetables, but not grain products, increased. Similar effects were observed at 12 and 18 months. Black and non-Hispanic white women had similar levels of reduction in fat, but the decrease in Hispanic women was less. Changes did not vary significantly by education. While bias in self-reported intakes may have resulted in somewhat overestimated changes in fat intake, the reported reduction was similar to the approximately 10% of energy decrease found in most trials and suggests that large changes in fat consumption can be attained in diverse study populations and in many subgroups. PMID- 10369505 TI - Blood transfusion and risk of non-Hodgkin's lymphoma. AB - A few epidemiologic studies have suggested that blood transfusion may be a risk factor for non-Hodgkin's lymphoma. The authors tested this hypothesis in a population-based, case-control study, using pathologically verified non-Hodgkin's lymphoma cases and transfusion documented via medical records. In 221 age- and sex-matched case-control pairs from Olmsted County, Minnesota, in 1975-1993, the authors observed an odds ratio of 0.84 (95% confidence interval 0.50-1.41) for history of transfusion and non-Hodgkin's lymphoma. There also was no apparent association between transfusion and non-Hodgkin's lymphoma in any subgroup analysis. Results do not support the hypothesis that blood transfusion contributes to the occurrence of non-Hodgkin's lymphoma. PMID- 10369506 TI - Neural tube defects along the Texas-Mexico border, 1993-1995. AB - In response to a 1991 anencephaly cluster in Cameron County, Texas, a surveillance and neural tube defect (NTD) recurrence prevention project for NTDs was implemented in the 14 Texas-Mexico border counties. For 1993-1995, NTD affected pregnancies were identified at all gestational ages through active surveillance of multiple case-ascertainment sources. There were 87 cases of anencephaly, 96 cases of spina bifida, and 14 cases of encephalocele for respective rates of 6.4, 7.1, and 1.1 per 10,000 live births. Of the 197 NTD case women, 93% were Hispanic. The overall, Hispanic, and Anglo NTD rates were, respectively, 14.6, 14.9, and 10.6 per 10,000 live births. The NTD rate for El Paso County (9.8 per 10,000), the most northwestern Texas county, was significantly lower (p = 0.001) than the aggregate rate for the rest of the Texas border (17.1 per 10,000). The overall Texas border rate was significantly higher (p < 0.001) than a recently estimated rate of 9.3 for California and minimally higher than a recently adjusted rate of 11.3 for the Metropolitan Atlanta Congenital Defects Program counties (p = 0.052), both of which now reflect all gestational ages. Of the 197 Texas border cases, 85% (168 cases) reached a gestational age of > or =20 weeks. Excluding cases of <20 weeks' gestation in the rate had a more marked effect on reducing the anencephaly rate (4.9 per 10,000) than the spina bifida rate (6.7 per 10,000). A country of birth was known for 153 (83%) of the 184 Hispanic case-women: 63% were born in Mexico; 24%, in Texas; and 11%, elsewhere in the United States. Rates for Mexico-born Hispanic women (15.1 per 10,000) were significantly higher than rates for United States-born Hispanic women (9.5 per 10,000) (p = 0.006). PMID- 10369507 TI - Impact of multiple births and elective deliveries on the trends in low birth weight in Norway, 1967-1995. AB - To describe trends in low birth weight (less than 2,500 g), the authors analyzed 1.7 million live births and stillbirths registered between 1967 and 1995 in the Medical Birth Registry of Norway. The proportion of low birth weight infants declined from 5.3% in 1967 to 4.5% in 1979 and was followed by a steady increase that reached 5.3% in 1995. Similar trends were observed in the proportion of preterm births. Mean birth weight increased from 3,456 g in 1967 to 3,518 g in 1995. From 1979 to 1987, the increase in the prevalence of low birth weight was related to single births, and after 1987 it was related to multiple births, which increased from 2.3% of all births in 1987 to 3.1% in 1995. The proportion of low birth weight in births occurring after 37 weeks of gestation declined continuously, resulting in low birth weight births' to an increasing extent being made up of births occurring before 37 weeks of gestation. In an ecologic analysis based on county of maternal residence, the increase in low birth weight among single births was accounted for by an increase in deliveries with induction of labor or cesarean section. The authors conclude that the overall proportion of low birth weight births is not a good indicator of health in a population with extensive use of obstetric procedures that affect gestational age or assisted fertilization, which increases the number of multiple births. PMID- 10369508 TI - Child mortality in Stockholm during 1885-1910: the impact of household size and number of children in the family on the risk of death from measles. AB - Previous studies have associated overcrowding at the household level with increased mortality, especially from airborne diseases. This association may be confounded by associations with adverse socioeconomic conditions or low age at infection. This study investigated the effect of crowding on the risk of measles death. Individual entries in a population-based register and on death certificates for children aged 0-15 years living in one parish in Stockholm in 1885, 1891, and 1910 (n = 36,718) were used to analyze cause-specific and overall death rates in relation to household size and the number of children in the household, using Cox regression analysis. Bivariate analysis identified significant relations between crowding and the cause-specific risk of death, which were subsequently tested while controlling for other known risk factors for childhood death. Significant negative associations between crowding and the risk of death from pneumonia and bronchitis disappeared when controlling for other risk factors. A negative association between the risk of overall death and large household size became significantly positive when controlling for other risk factors. The increased risk of death from measles associated with proxies for crowding remained after controlling for other risk factors. In conclusion, crowding may have a statistically independent effect on the risk of death from measles. PMID- 10369509 TI - Puumala virus infections in Finland: increased occupational risk for farmers. AB - Puumala hantavirus, transmitted by bank voles (Clethrionomys glareolus), causes a mild-type hemorrhagic fever with renal syndrome. The disease is common in Finland and is considered an occupational hazard for farmers, but the actual risk has not been assessed by analytical studies. Data on 5,132 serologically confirmed Puumala virus infections during 1989-1994 were analyzed, and cases among farmers and the population living in similar conditions were compared. The farmers contracted the disease earlier and more often than did the comparison group. In the province of Mikkeli with the highest incidence (70/100,000), the risk ratio was 5.1 (95% confidence interval (CI) 3.0-8.4) for 20- to 29-year-old farmers; in the older age groups, the risk was still increased but the risk ratios were lower. The peak incidence in the comparison group was 10 years later (age group 30-39 years). For the whole country, the result was similar although less marked. The average risk ratio adjusted by age, sex, and geographic variation was 1.7 (95% CI 1.5-1.8) for the whole country and 1.9 (95% CI 1.5-2.3) for the Mikkeli province, where 80% of Puumala virus infections among young farmers could be estimated to be attributable to occupation. PMID- 10369510 TI - Effects of temperature and snowfall on mortality in Pennsylvania. AB - The relation between exposure to severe cold weather and mortality is examined in a retrospective study of deaths occurring during the month of January from 1991 to 1996 in Pennsylvania. Using division-days as units of observation (n = 1,560) aggregated from death certificates and geographic divisions, the authors estimated mortality rates for total deaths and deaths due to ischemic heart disease, cerebrovascular diseases, and respiratory diseases by analyses based on generalized estimating equations. Total mortality increased on days of "extreme" climatic conditions, that is, when snowfall was greater than 3 cm and when temperatures were below -7 degrees C (rate ratio (RR) = 1.27, 95 percent confidence interval (CI) 1.12-1.44). On days of extreme conditions, mortality due to ischemic heart diseases tripled among males aged 35-49 years (RR = 3.54, 95 percent CI 2.35-5.35), increased for men aged 50-64 years (RR = 1.77, 95 percent CI 1.32-2.38), and rose for males aged 65 years and older (RR = 1.58, 95 percent CI 1.37-1.82), when compared with milder conditions. Among females, mortality for those aged 65 years and older increased for respiratory causes (RR = 1.68, 95 percent CI 1.28-2.21) and cerebrovascular causes (RR = 1.47, 95 percent CI 1.13 1.91). Cold and snow exposure may be hazardous among men as young as 35 years. PMID- 10369511 TI - Past, present, and future directions for Alzheimer research. AB - To understand the current thinking and future research directions in Alzheimer disease (AD), it is essential to examine the breadth of research into AD. AD was identified as a distinct disease process early in this century and research into its etiology and treatment has a short, but active, history. This paper emphasizes past, present, and future biomedical research for the goal of stimulating research to solve some of the unanswered questions to ultimately help the victims of Alzheimer disease. The "hot" topics today for biomedical researchers include early diagnosis, presymptomatic markers, the role of presenilins, amyloid, protein precursor processing, and continued exploration for genetic and environmental risk factors. PMID- 10369512 TI - Alzheimer disease 1999: a status report. AB - Although the cause of Alzheimer disease (AD) remains a mystery, a great deal of progress has been made over the past 15 years. A large body of evidence suggests that beta amyloid (Abeta) deposition and toxicity are critical steps leading to neuronal death in AD. Evidence is beginning to show that antioxidant and antiinflammatory drugs and estrogen may prevent, delay onset, or slow the progression of AD. Until a cure is found it will be necessary to provide high quality long-term and palliative care in the community or in specialized treatment centers. The AD patient's caregiver often suffers in silence and research is needed to better understand disease burden to design appropriate interventions for these victims of AD. PMID- 10369513 TI - Building and maintaining an interdisciplinary research team. AB - Successful grant-writing in Alzheimer disease research, as with many other diseases, requires collaborative work among a group of individuals who represent various disciplines of relevance to the research problem. Interdisciplinary teams in Alzheimer disease research have the potential to explore more facets of a given research problem than teams that are not interdisciplinary in nature. Such teams also have the potential to produce better science and to disseminate results to a wider spectrum of relevant groups, may be more successful in achieving funding, and are stimulating and growth-enhancing for members. Building and maintaining these teams is a complex and challenging process, but identification and proactive resolution of challenges is essential. Important elements of success include establishing common goals, using a democratic group process, maintaining open communication, developing mutually acceptable policies for disseminating research results, and facilitating achievement of team members' personal and professional goals. PMID- 10369514 TI - Selection, care, and feeding of a research mentor. AB - The purpose of this article is to describe a process by which investigators can identify and select a research mentor and develop a mutually beneficial mentoring relationship. Such a relationship can benefit not only novice investigators, but also more experienced researchers who are branching out into a new field of inquiry. The term mentoring is defined and nature of the mentoring relationship is discussed. Specific information is provided regarding the types of assistance a research mentor can provide, sources of potential mentors, qualities of the successful mentor and protege, stages of the mentoring relationship, and strategies for dealing with less than optimal relationships. Tools are provided to aid the reader in identifying research needs and in rating potential mentors. PMID- 10369515 TI - Gaining and sustaining minority participation in longitudinal research projects. AB - African-Americans are often not enthusiastic about participating in Alzheimer disease research due to past exploitation from medical and public health studies. To assure adequate representation from the African-American community, and to investigate the health needs of this population, strategies to recognize the problems and address the issues have been developed. The recruitment approach used existing connections in an urban community using culturally sensitive strategies. These strategies recruited elderly patients from an existing relationship with the Boston University Medical Center Geriatric Service. They built on the researchers' association with a neighborhood health center located in one of the Boston communities and utilized the services of an outreach worker. The outreach worker was from the community, aware of the health care needs of the elderly, and had a relationship with existing elderly community groups. Several methods were used to recruit patients. These included one-on-one discussions, posters, existing relationships with elderly groups, and direct phone calls. To develop trust carefully with the community's elders, multiple trust-building strategies were used. A multifaceted recruitment approach and strong linkages between the research team and the community were essential to successful recruitment and retention of participants. PMID- 10369516 TI - Conducting research with urban elders: issues of recruitment, data collection, and home visits. AB - The National Institutes of Health 1994 guidelines require inclusion of minorities in research. People seeking grants need to be aware of techniques for recruitment of minority populations. The Boston University Alzheimer's Disease Center addresses difficult recruitment issues that arise from a skeptical minority population. The approach uses home visits to circumvent many of the barriers to recruitment and retention of participants in research studies. PMID- 10369517 TI - Brain banking: basic science methods. AB - The fundamental objectives of a brain bank are to document precisely the gross anatomical and histological findings, to establish accurate neuropathological diagnoses using well-defined criteria and standardized dissection techniques, and to serve as a source of fresh, fixed, and deep-frozen brain tissue. The brain tissue is optimally prepared and stored to provide high-quality research material that is suitable for a wide variety of investigations. A computerized database is used to classify all clinical and pathological diagnoses, neuropathological data, cause of death, postmortem interval, associated chronic diseases, and storage conditions of the tissue. Tissue availability and distribution is monitored and quality assurance is provided regarding tissue acquisition, processing, storage, and diagnosis. These functions facilitate clinicopathological correlative studies, neurochemical, molecular biological, immunohistochemical, quantitative and in vitro studies. Furthermore, the use of standardized tissue methods promotes multicenter collaborations. PMID- 10369518 TI - Qualitative research and Alzheimer disease. AB - This article presents an overview of qualitative research methods and illustrates ways in which these methods may be useful in discovering unknowns that contribute to knowledge about Alzheimer disease. An observation of an elder with Alzheimer disease sets the scene for thinking about ways to explore, describe, and interpret human experiences and the context within which these experiences are perceived. The philosophical assumptions and standards for qualitative research methods are discussed. Two of the qualitative research approaches (ethnography and grounded theory) are presented using examples from published studies to illustrate the logical consistency of fit between purpose statement, data collection strategies, analysis structures, and results specific to the language, and guidelines and rules for each of these two approaches. Practical issues of data management, including the availability of qualitative computer packages and publication, are addressed briefly. PMID- 10369519 TI - Quantitative research methods. AB - This paper provides some ideas for improving the quality of Alzheimer disease (AD) research projects by using the appropriate quantitative methods. Statistical tests for answering research questions are presented according to the types of design used. Recently published research studies illustrate some pitfalls to avoid and issues to consider when planning research; threats posed by multiple comparisons, how to control for threats, and cautions to use in interpreting those studies that do not control for the effects of chance. Careful attention to these details will strengthen the work and thus improve the state of the science relative to the care of persons with Alzheimer disease. PMID- 10369520 TI - Ethical issues in Alzheimer disease research. AB - Research in Alzheimer disease (AD) is required to advance knowledge for the ultimate goal of finding a cure or prevention and to improve care for persons with AD and their caregivers. All research needs to be planned and conducted ethically, but research in AD has many ethical issues to consider due to the diminished cognitive capacity of potential subjects and genetic testing for AD. This paper describes the development of guidelines for ethical research, applying ethical principles to issues in human subjects research, and ethical issues in AD research. Important ethical questions must be addressed in AD research and basic moral and professional values will certainly be tested as the state of the science improves and research on AD, especially research involving genetic testing for the disease, becomes more acceptable and available. PMID- 10369522 TI - Making a positive difference in the lives of nursing home residents with Alzheimer disease: the lifestyle approach. AB - Many persons with Alzheimer disease (AD) receive care outside their home, temporarily at a day care center or permanently in an assisted living arrangement or in a skilled nursing facility. How persons with AD live, and the potential to improve the lives of persons with AD, is a rich resource for conducting interdisciplinary research in AD. This paper presents the "lifestyle approach" which is a method of easing the transition from his/her home into a nursing home setting. The lifestyle approach helps establish a daily routine for residents that is familiar and that fills their days with meaningful activities. Little research has been initiated to study how to increase the quality of life of nursing home residents with AD. Each component of the lifestyle approach is presented and suggestions for studies to support or refute the author's opinion are made to stimulate research in this area. PMID- 10369521 TI - Assessment of behavioral symptom management in demented individuals. AB - This article is intended to stimulate interdisciplinary research by social, behavioral, and biomedical researchers to solve some of the problems for persons with Alzheimer disease who suffer from behavioral symptoms. We identify the need for using conceptually based measures that have adequate reliability and validity to operationalize indicators of psychological well-being, suggest instruments to assess mood, engagement, agitation, resistiveness and comfort, and pose research questions to explore responses to active involvement (individual or group), passive involvement, lack of activity, aversive activity (related to activities of daily living), and limitation of activity (restraints) in persons with dementia. Answers to the research questions posed in this article will lead to expanded knowledge and can redefine how we give care to those in need. PMID- 10369523 TI - Nursing research and solving problems of Alzheimer disease. AB - This article discusses the applied science of the nursing discipline and nursing issues related to Alzheimer disease. Three models are presented and illustrated with interdisciplinary research ideas to improve the care of persons who suffer from dementia of the Alzheimer type (DAT). Phases of Nursing Research is a five component model to (1) identify problems, (2) conduct the study, (3) disseminate results, (4) utilize findings that are to the central component, (5) practical knowledge. Promoting evidence-based care for making a recommendation against artificial feeding and providing the tools to promote natural feeding illustrates the Interdisciplinary Collaboration model. The Program of Research model is described by studies of late-stage DAT, intercurrent infections, and decision making. Issues framing nursing research are discussed and specific suggestions for research projects developed from recent funding priorities of the National Institutes of Health are given. PMID- 10369525 TI - The biochemistry of Alzheimer disease. AB - Biochemical and genetic investigations have identified four key proteins, mutations in which either cause Alzheimer disease (AD) (beta-amyloid precursor protein, presenilin 1 and 2) or confer a higher risk of developing AD (apolipoprotein E). This paper discusses the biochemical evidence that links each protein to AD, various animal and cell models that have been used in these investigations, and the putative interactions between these proteins that lead to AD. Areas that are especially fertile for novel research are noted as are gaps in our present understanding of the etiology of AD. PMID- 10369524 TI - Future directions in the analysis of gene expression in Alzheimer disease. AB - This review briefly explores future directions for gene expression research in Alzheimer disease (AD). Two complementary techniques, differential display and gene expression arrays, allow researchers to identify differentially expressed genes. These techniques promote a systematic examination of gene expression. In conjunction with a technique that allows the isolation of mRNA from individual cells, differential display and gene expression array technology can be used to examine AD-induced changes in gene expression patterns in individual cells. This area of research should elucidate information that will provide a greater understanding of the pathological processes in AD and aid in the development of therapeutic agents. PMID- 10369526 TI - Social work perspectives: issues in caregiver research: the family. AB - More than 4 million persons in the United States are believed to have Alzheimer disease. Seven of 10 are cared for in the community by family members who make numerous decisions ranging from those that have to do with personal care and daily activities to participation in research. Families who do participate in research or who agree for the dementia-impaired individual to participate, do so for a variety or reasons, including the altruistic reason of finding a cure or treatment that may help others. This paper looks at the caregiving experience and those issues that may influence the family to participate in research, what families expect from research, and how they expect researchers to treat the impaired relative. PMID- 10369527 TI - Trends in the content and methodology of Alzheimer caregiving research. AB - Research into the experience of caregiving for patients with Alzheimer disease continues to grow steadily both in the number of studies and the scope of investigation. A review of the current trends in the content and methodology of research into caregiving is provided as a guide for new investigators entering this field. Whereas most studies continue to focus on the stress and burden associated with caregiving, increased study into topics such as treatment utilization, clinical decision-making, and caregivers and early detection will continue to broaden and enrich the scope of investigation. Positive trends in research methodology, including improved sampling procedures, more frequent inclusion of control groups, the use of improved measurement techniques, and the use of more sophisticated designs and statistics, will all continue. Despite the many contributions of past research into caregiving, future studies promise to make a critical contribution to a number of fields of study. PMID- 10369528 TI - The Health Outreach Program for the Elderly (HOPE) database. AB - The purpose of the Boston University Alzheimer's Disease Center's Health Outreach Program for the Elderly (HOPE), is to facilitate research to solve the problems of Alzheimer disease (AD). The HOPE project has supported the development of a database that can be made available to qualified researchers who are studying AD. This article provides a brief introduction to databases and database concepts. In addition, the database created by the Boston University Alzheimer's Disease Center is described. The structure of the database is presented and the datapoints it contains are described. The procedures for requesting a database query are outlined. PMID- 10369529 TI - It all starts with an idea. AB - Great ideas make great research and provide the researcher with the shapes for growing the idea in an organized fashion. Successful research ideas are interesting, important, researchable, and doable. This article describes one way of organizing great ideas into successful research by using the research process, beginning with conceptualization/design and literature review, and proceeding through the articulation of the framework, purpose, assumptions, limitations, definitions, and hypotheses/research questions. Following the logical progression of the conceptualization of the idea will help the researcher to complete the grant application and be ready to develop and execute the methodology of the study. PMID- 10369530 TI - Idea to application: you are in charge. AB - The grant writing process can be daunting to a novice investigator. This article focuses on three inter-related activities that lead to a completed grant application. The first activity involves developing the research team. Selection of colleagues who will strengthen and support the research ideas is an early first step to developing a competitive grant application. It is important to communicate clearly the roles and responsibilities of each team member. The second activity is development of the scientific plan-the research proposal-from specific aims to plans for data analysis. Strategies used to review the literature, synthesize existing knowledge, develop the argument for the research proposal, and select the most appropriate method are discussed. The third activity involves developing a business plan-the budget-that demonstrates clearly the investigator's ability to complete the proposed research. These three activities are set within a timeline to guide investigators in successful completion of a grant application. PMID- 10369531 TI - Structure and process of federal funding for AD research. AB - This paper outlines the structure of the National Institute on Aging (NIA), the National Institutes of Health (NIH) agency that funds a majority of Alzheimer disease (AD) research, and the process of submitting a federal grant application. The multiple steps of the grant application process, both for the investigator preparing the application and the review process at NIH, are described. The importance of speaking with a program officer from an institute and incorporating reviewers' comments into grant applications to strengthen the proposal is stressed. PMID- 10369532 TI - The grant application: making yours stand out across the review cycle. AB - This article provides an overview of the grant application during the review process. Three levels of review occur once a grant is submitted-administrative, scientific, and programmatic. The purposes of each level of review are different, but the overall goal for the funding agency is to fund grant applications that address important research topics, are scientifically meritorious, and assist the agency in meeting their goals and objectives. PMID- 10369533 TI - Publishing the research report. AB - Although research is not finished until it is published, many investigators fail to report their research. The purpose of this article is to discuss the dissemination of research findings via the published manuscript. Barriers to publication are reviewed and strategies for overcoming these barriers are presented. Selection of a target journal is discussed, along with the purpose of the query letter. The format of the research report is addressed and specific information is provided regarding the literature review and the use of illustrative materials. Practical tips are provided for each stage of the writing process. PMID- 10369534 TI - Androgenetic alopecia. PMID- 10369535 TI - Diffuse hair loss. PMID- 10369536 TI - Alopecia areata: diagnosis and management. PMID- 10369537 TI - Congenital hypotrichosis. PMID- 10369538 TI - Traumatic alopecia. PMID- 10369539 TI - Leprosy: pathogenesis updated. PMID- 10369540 TI - Actinic reticuloid. PMID- 10369541 TI - The art, the science, and the practice of dermatology in the next millennium. PMID- 10369542 TI - Lesional T-cell subset in leprosy and leprosy reaction. AB - BACKGROUND: The T-cell-mediated immune response plays an important role in leprosy. The in situ proportion and pattern of distribution of T-cell subsets in leprosy skin lesions have been studied, but no conclusion could be drawn. METHODS: We used monoclonal antibodies for T-helper and T-suppressor surface antigen to define the nature of dermal infiltration in 17 cases of nonreactional leprosy and 20 cases of reactional leprosy. RESULTS: We found T helper admixed with T suppressor in an aggregated pattern in the granulomas of most cases of nonreactional leprosy and in type I reactional leprosy, but a diffuse infiltrate throughout the dermis of type II reactional leprosy. The T-helper/suppressor ratio was 1.68 in tuberculoid and 1.5 in lepromatous cases. The T-helper/ suppressor ratios of borderline tuberculoid (3.11) and type I reactional leprosy (2.54) were not statistically different. The T-helper/suppressor ratio of type II reactional leprosy (5.83) was statistically higher than nonreactional lepromatous cases. CONCLUSIONS: The alteration of the T-helper/suppressor ratio in our study is mainly due to the reduction of T-suppressor cells in the dermal infiltrates, especially in type II reactional leprosy. Further studies of T-suppressor functions may be important in the pathogenesis of leprosy. PMID- 10369543 TI - Pruritic papular eruption in HIV seropositive patients: a cutaneous marker for immunosuppression. AB - BACKGROUND: Previous reports have shown the correlation between certain skin disorders and immune status in human immunodeficiency virus (HIV) infected patients. Pruritic papular eruption (PPE) is the most common cutaneous manifestation in HIV infected patients. The purpose of this study is to define the relationship between the presentation of PPE and the immune status in HIV infection, as measured by the T-cell subset, and to establish the usefulness of this common eruption as a predictor of CD4 count. METHOD: In this cross-sectional study, 20 HIV-positive patients with characteristics of PPE were studied. Clinical data, skin biopsy, and immune status, evaluated by measuring CD4, CD8, and CD4/CD8, were investigated. RESULTS: Seventy-five per cent of patients already had antecedent skin disorders, so PPE is not a leading symptom in HIV infected patients; 81.25% of PPE patients had an advanced degree of immunosuppression with a CD4 count below 100/mm3 and 75% below 50/mm3. CONCLUSIONS: PPE can be regarded as a cutaneous marker of advanced HIV infection. PMID- 10369544 TI - Pemphigus in Kuwait. AB - BACKGROUND: Although pemphigus is a well-characterized entity, detailed epidemiologic studies from the Arabian Peninsula are not available. The purpose of this study was to elucidate the clinical features, course, and prognosis of pemphigus patients followed at a national dermatology center in Kuwait. METHODS: Fifty-four patients with pemphigus in this report were treated between 1981 and 1996, and were studied for several clinical features, treatment, course and prognosis. RESULTS: Around 80% of pemphigus patients were Arabs, and Kuwaitis constituted the largest number (46.3%) with a female predominance (F: M = 2:1). Pemphigus vulgaris (PV) was the commonest clinical type. The mean age of onset was 36 years. The follow-up period ranged from 2 months to 12 years (mean, 4.5 years). The majority of the patients could be managed with low-dose steroids (30 60 mg/day). Twenty per cent of the patients were in complete clinical remission and were off systemic therapy for an average of 3 years. No death secondary to the disease or its treatment was observed. CONCLUSIONS: Kuwaiti patients with pemphigus were observed to have a relatively young age of onset and a female predominance. Low doses of steroids were enough to control the disease in the majority, and at least 20% of patients were off therapy and in complete remission on follow-up. PMID- 10369545 TI - A survey of reference accuracy in two Asian dermatologic journals (the Journal of Dermatology and the Korean Journal of Dermatology). AB - BACKGROUND: The reference list is an important part of a scientific article. To be useful, it must be accurate. METHODS: To evaluate the reference accuracy in the Journal of Dermatology and the Korean Journal of Dermatology, we randomly selected 100 references from each journal and checked them against the original articles. RESULTS: The overall rate of citation errors was 24% in the Journal of Dermatology and 33% in the Korean Journal of Dermatology. Errors in the title and author names were common, each occurring in about half of the citation errors. The overall rate of quotation errors was 14% in the Journal of Dermatology and 27% in the Korean Journal of Dermatology. CONCLUSIONS: This study shows that the rate of citation errors is unacceptably high in the Journal of Dermatology and the Korean Journal of Dermatology, which significantly diminishes the value of the reference list. We would strongly urge that the peer review of citation and quotation accuracy should be strengthened. PMID- 10369546 TI - Skin pathology findings in a cohort of 1500 adult and elderly subjects. AB - BACKGROUND: No extensive studies are available in the literature on the eventual skin pathology induced by neurologic or systemic diseases in elderly individuals. Other factors, such as health and hygiene, socioeconomic status, and climate can also play an important role. METHODS: Fifteen-hundred subjects (886 women and 614 men; mean age, 67.8 years; range, 39-90 years) were admitted to the Department of Geriatrics at the Oasi Institute between 1992 and 1997; all these subjects were carefully evaluated from a dermatologic point of view. Each subject underwent specialist examinations, routine blood analyses, thoracic X-rays, cerebral computerized tomography (CT) scan, and magnetic resonance imaging (MRI) when appropriate. A group of subjects without significant neurologic or systemic disease, comprising 116 women and 60 men (mean age, 64.5 years; range, range, 40 90 years), was selected and used as a normal control group. Subsequently, our attention was focused on the eventual presence of the following neurologic diseases: Alzheimer-type dementia, vascular dementia, mixed-type dementia, subcortical dementia, Parkinson's disease, vascular brain disease, hemiplegia, etc. Thus, different subgroups were formed on the basis of such diagnostic categories and the frequency of skin pathology in each subgroup was evaluated. RESULTS: Of the 1500 subjects, 1439 stated that they had never been affected by dermatologic disease. No statistically significant difference in frequency of skin pathology was found between normal controls and the different patient subgroups. Unsuspected and singular dermatoses were found, however, such as paraneoplastic syndromes, idiopathic tripe palms, white fibrous papulosis of the neck as an expression of photoaging, conditions induced by former popular traditions of Sicilian culture (anetoderma secondary to the application of Hirudo medicinalis and erythema ab igne), pigmented dermatoses never described before in Italy (prurigo pigmentosa and friction amyloidosis), and nail abnormalities (atypical half-and-half nail, and dyschromic nail changes in multiple system atrophy and in hemiplegia). CONCLUSIONS: The dermatologic screening performed in 1500 patients revealed several unexpected diagnoses and some original observations. Some rare dermatoses were described and certain hypotheses were suggested to explain the peculiar dyschromic changes of the fingernails in multiple system atrophy, the atypical cases of half-and-half nail, and the so called idiopathic tripe palms associated with psoriasis. PMID- 10369547 TI - Carcinoma erysipeloides as a presenting feature of breast carcinoma. PMID- 10369548 TI - Eosinophilic fasciitis responsive to treatment with pulsed steroids and cyclosporine. PMID- 10369549 TI - Pityriasis lichenoides and acquired toxoplasmosis. PMID- 10369550 TI - Primary squamous syringometaplasia with no underlying malignancy. PMID- 10369552 TI - Recombinant human granulocyte-macrophage colony-stimulating factor applied locally in low doses enhances healing and prevents recurrence of chronic venous ulcers. AB - BACKGROUND: Chronic venous leg ulcers have a major medical and economic impact on the elderly worldwide. Healing of the large ulcers (>10 cm2) occurs only in two thirds of the patients and reulceration of healed ulcers recurs in one-third within 1 year. Because both healing and relapse rate influence greatly a patient's quality of life and the overall cost of treatment, every effort should be made to improve these two parameters. OBJECTIVE: To determine the safety and efficacy of topical low-dose recombinant human granulocyte-macrophage colony stimulating factor (rhu GM-CSF) for the treatment of venous ulcers, and to document any improvement in healing rates. METHODS: Thirty-eight patients (29 women, 9 men; median age, 74 years) with chronic venous insufficiency were treated with topical rhu GM-CSF (5 microg/mL 0.9% sodium chloride solution), followed by application of a compression dressing. All subjects were treated as outpatients. RESULTS: Complete healing was observed in 47 of the 52 ulcers (90.4%). The average healing time was 19 weeks. No systemic or local side-effects from the therapy were observed. Nine chronic ulcers, previously refractory to conventional treatment (pretreatment for more than 46 weeks), showed the same response rate (9/8, or 88.9%) and healing time (mean, 19 weeks). After 40 months, no reulceration of the healed ulcers was observed, but two patients developed new ulcers on the same leg. Healing remained stable, with excellent cosmetic results. CONCLUSIONS: In this first study, topically applied low-dose rhu GM-CSF was a safe treatment for chronic venous leg ulcers. Healing rates were significantly increased and relapse rates were minimal. PMID- 10369551 TI - Terbinafine vs. placebo for onychomycosis in black patients. PMID- 10369553 TI - A controlled trial of traditional Chinese herbal medicine in Chinese patients with recalcitrant atopic dermatitis. AB - BACKGROUND: There have been published reports from the United Kingdom of good responses to the use of traditional Chinese herbal medicine (Zemaphyte, Phytopharm Plc, Cambridge, UK) in treating recalcitrant atopic dermatitis. We conducted a double-blind, placebo-controlled, cross-over study among Chinese patients with recalcitrant atopic dermatitis using this same herbal preparation. METHODS: Forty patients were recruited. They were given Zemaphyte and placebo in random order, each for 8 consecutive weeks with a 4-week wash-out period in between. Scores based on the severity and extent of four clinical parameters (erythema, surface damage, lichenification and scaling) were recorded at baseline and at 4-weekly intervals throughout the 20-week trial period. RESULTS: Thirty seven patients completed the trial. There was a general trend of clinical improvement with time throughout the trial period in both patient groups, irrespective of whether they received Zemaphyte or placebo first. Zemaphyte, however, offered no statistically significant treatment effect over placebo for all four clinical parameters, except for lichenification at week 4. There were no significant carry-over effects. Blood tests for hematologic, renal and liver functions were all normal throughout the trial. CONCLUSIONS: Zemaphyte did not seem to benefit Chinese patients with recalcitrant atopic dermatitis in our study. Further research is required to evaluate its efficacy. PMID- 10369554 TI - Density of viral particles in pre and post Nd: YAG laser hyperthermia therapy and cryotherapy in plantar warts. AB - BACKGROUND: Warts often present a difficult treatment problem for clinicians because of the lack of specific antipapillomavirus agents. Plantar warts, in particular, represent a therapeutic challenge. METHODS: Twenty-five patients with plantar warts were treated with Nd:YAG hyperthermia and another 25 were treated with cryotherapy. Biopsies were taken before and after treatment in both groups and were examined for the presence of human papillomavirus deoxyribonucleic acid (HPV DNA) using in situ hybridization (ISH). RESULTS: HPV DNA was detected in 100% of untreated warts and in 96% of cryotreated warts, but was not detected in any of the hyperthermia-treated warts. CONCLUSIONS: HPV is more vulnerable to hyperthermia than to cryotherapy. PMID- 10369555 TI - Neuroprotective effects of basic fibroblast growth factor following spinal cord contusion injury in the rat. AB - Cytokines and neurotrophic factors have been implicated in the pathophysiology of injury to the central nervous system. While some cytokines are considered pro inflammatory, other factors promote neuronal growth and survival. The present study investigated the neuroprotective effects of interleukins 1 (IL-1), 4 (IL 4), and 6 (IL-6), nerve growth factor (NGF), ciliary neurotrophic factor (CNTF), and basic fibroblast growth factor (bFGF) in a contusion model of spinal cord injury. Female Sprague-Dawley rats (n = 55) sustained a 10-g weight-drop injury to the lower thoracic spinal cord (T10) from a height of 12.5 mm using the NYU impactor. A micro-infusion system (Alzet minipump) was used to continuously deliver drugs or vehicle directly into the epicenter of the contused spinal cord starting 1 or three h postinjury. At the end of 7 days, animals were perfused and the cords removed for histopathological analysis. Longitudinal serial sections were cut on a freezing microtome and stained with cresyl violet. Areas of central necrosis, partial preservation, and total zone of tissue injury were identified and traced by an independent reviewer using a computer based imaging system. The mean total zone of injury in five animals receiving vehicle infusion was 18.04+/ 4.20 mm3. The mean zone of partial preservation in these animals was 16.46+/-3.32 mm. Basic fibroblast growth factor reduced the total zone of injury by 33% [p<0.01, least significant difference (LSD) of Fisher] in five animals and the zone of partial preservation by 32% (p<0.01, LSD of Fisher) when compared to controls. There were trends toward reduction in total zone of injury and zone of partial preservation in rats treated with IL-4, CNTF, and NGF versus vehicle; however, none of these reached statistical significance. No significant differences were observed between animals receiving vehicle versus bFGF treatment commencing 3 h after injury. These data demonstrate that the continuous intramedullary infusion of bFGF initiated one hour after moderate contusion injury of the spinal cord significantly reduces the total zone of injury and the zone of partial preservation. These results support the further investigation and possible future clinical application of bFGF in the treatment of acute spinal cord contusion injury. PMID- 10369556 TI - Site-specific cleavage of 28S rRNA as a marker of traumatic brain injury. AB - A cleavage product of 28S rRNA was isolated from ipsilateral hippocampus of rat brain subjected to lateral fluid percussion induced traumatic brain injury (TBI). Northern blot analysis demonstrated that the corresponding cDNA fragment hybridized to 28S rRNA and three cleavage products. Two of the cleaved rRNA fragments (1.3 kb and 0.9 kb) were also observed in differentiated PC12 cells undergoing apoptosis induced by NGF withdrawal. The third fragment (0.6 kb) was detected only in rat brain tissue subjected to trauma, suggesting specific cleavage of 28S following TBI. The 0.6-kb fragment was found only in cortex and hippocampus ipsilateral to the trauma site, but not in brain stem, contralateral cortex or contralateral hippocampus. 28S rRNA cleavage was detected beginning 2 h after trauma and reflected injury severity. Although cleavage of 28S rRNA has been reported in association with apoptosis in white blood cells and apoptosis occurs in the experimental head injury model used, the pattern of 28S rRNA cleavage observed with TBI differs from those observed in apoptotic PC12 cells or those reported for white blood cells. Thus, whereas 28S rRNA fragmentation appears to be a marker of posttraumatic brain injury, its precise role in the secondary injury process remains to be established. PMID- 10369557 TI - Closed head injury in the rat induces whole body oxidative stress: overall reducing antioxidant profile. AB - Traumatic injury to the brain triggers the accumulation of harmful mediators, including highly toxic reactive oxygen species (ROS). Endogenous defense mechanism against ROS is provided by low molecular weight antioxidants (LMWA), reflected in the reducing power of the tissue, which can be measured by cyclic voltammetry (CV). CV records biological peak potential (type of scavenger), and anodic current intensity (scavenger concentration). The effect of closed head injury (CHI) on the reducing power of various organs was studied. Water and lipid soluble extracts were prepared from the brain, heart, lung, kidney, intestine, skin, and liver of control and traumatized rats (1 and 24 h after injury) and total LMWA was determined. Ascorbic acid, uric acid, alpha-tocopherol, carotene and ubiquinol-10 were also identified by HPLC. The dynamic changes in LMWA levels indicate that the whole body responds to CHI. For example, transient reduction in LMWA (p<0.01) in the heart, kidney, lung and liver at 1 h suggests their consumption, probably due to interaction with locally produced ROS. However, in some tissues (e.g., skin) there was an increase (p<0.01), arguing for recruitment of higher than normal levels of LMWA to neutralize the ROS. alpha-Tocopherol levels in the brain, liver, lung, skin, and kidney were significantly reduced (p<0.01) even up to 24 h. We conclude that although the injury was delivered over the left cerebral hemisphere, the whole body appeared to be under oxidative stress, within 24 h after brain injury. PMID- 10369558 TI - Behavioral responses of C57BL/6, FVB/N, and 129/SvEMS mouse strains to traumatic brain injury: implications for gene targeting approaches to neurotrauma. AB - Recent studies have suggested that mouse models of traumatic brain injury may be useful for evaluating the role of single gene products in brain trauma. In the present study, we report that three background strains (C57BL/6, FVB/N, and 129/SvEMS), commonly used in genetically altered mice, exhibit significantly different behavioral responses when subjected to sham surgery (n = 9 per group) or moderate controlled cortical impact (CCI) injury (n = 12 per group). Injured animals from all three strains showed delayed recovery of pedal withdrawal and righting reflexes compared to sham-operated controls. Significant deficits in both a forepaw contraflexion and rotarod task were evident for up to 7 days after injury, with no significant difference among strains. Sham-operated C57BL/6 mice performed significantly better than FVB/N and 129/SvEMS sham controls in a beam walking task up to 4 weeks after surgery. However, CCI-injured FVB/N mice outperformed injured animals from both other strains in this same task. Significant impairment of place learning in the Morris water maze and Barnes circular maze was observed at 7-10 days and 21-24 days after injury, respectively, in C57BL/6 mice when compared with sham controls. Sham-operated FVB/N and 129/SvEMS mice were unable to learn either task, and performance did not differ significantly from respective CCI injured animals. Our results suggest that background strain should be carefully considered with experiments involving genetically altered mice, especially when planning behavioral outcome measures after CNS injury. PMID- 10369559 TI - Age-dependent impairment of K(ATP) channel function following brain injury. AB - Previous studies observed that endothelin-1 (ET-1) contributed to ATP-sensitive K+ (K(ATP)) channel impairment 1 h following fluid percussion brain injury (FPI) in the newborn pig. The present study was designed to determine the effect of FPI on K(ATP) channel activity as a function of time in newborn (1-5 days old) and juvenile (3-4 weeks old) pigs equipped with a closed cranial window. FPI of moderate severity (1.9-2.1 atm) was produced by using a pendulum to strike a piston on a saline-filled cylinder that was fluid coupled to the brain via a hollow screw inserted through the cranium. Cromakalim, a K(ATP) agonist, produced dilation that was blunted for at least 72 h post FPI, but dilator responsiveness was restored within 168 h post FPI in the newborn pig (15+/-1% and 27+/-2% vs. 5+/-1% and 11+/-1% vs. 13+/-1% and 26+/-2% for responses to 10(-8), 10(-6) M cromakalim before, and 72 and 168 h after FPI). Similar inhibited responses were observed for calcitonin gene-related peptide, 8-Bromo cGMP, and the nitric oxide (NO) releasers SNP and SNAP. In contrast, cromakalim-induced dilation was blunted for at least 4 h, but dilator responsiveness was restored within 8 h post FPI in the juvenile pig (15+/-1% and 27+/-1% vs. 9+/-1% and 15+/-2% vs. 18+/-1% and 28+/ 1% for 10(-8), 10(-6) M cromakalim before, and 4 and 8 h post FPI). Similar inhibition of dilations of other agonists also occurred in the juvenile. CSF ET-1 increased to a greater level and remained elevated for a longer period of time in the newborn compared to the juvenile pig. BQ123, an ET-1 antagonist, pretreatment partially restored decremented agonist induced dilation following FPI in the newborn and juvenile pig (5+/-1% and 11+/-1% vs. 11+/-1% and 21+/-1% for responses to 10(-8), 10(-6) M cromakalim 72 h post FPI in the newborn in the absence and presence of BQ123). These data indicate that K(ATP) channel function is impaired to a greater extent and for a longer time period in the newborn versus the juvenile pig. These data also show that ET-1 contributes to such impaired vascular responsiveness to a greater extent in the newborn versus the juvenile pig. These data furthermore suggest that the newborn is more sensitive to traumatic vascular injury than the juvenile. PMID- 10369560 TI - Terminally differentiated human neurons survive and integrate following transplantation into the traumatically injured rat brain. AB - The present study evaluated the survival and integration of human postmitotic neurons (hNT) following transplantation into the traumatically injured rodent brain. Anesthetized male Sprague-Dawley rats (n = 47) were subjected to lateral fluid percussion brain injury of moderate severity (2.4-2.6 atm). Sham animals (n = 28) were surgically prepared, but did not receive brain injury. At 24 h following injury or sham surgery, the rats were re-anesthetized and approximately 100,000 hNT cells (freshly cultured or previously frozen) or vehicle were stereotactically injected into the ipsilateral cortex. Animals were examined for neuromotor function at 48 h, 7 days, and 14 days posttransplantation using a standard battery of motor tests. Animals were sacrificed at 2 weeks postinjury and viability of hNT grafts was assessed by Nissl staining and MOC-1 immunohistochemistry, which recognizes human neural cell adhesion molecules (NCAM) expressed on hNT cells. Transplanted hNT grafts remained viable in 83% of brain-injured animals at 2 weeks following transplantation of either fresh or frozen hNT cells. Glial fibrillary acidic protein (GFAP) immunohistochemistry revealed a marked increase in the number of reactive astrocytes following brain injury in both vehicle and hNT implanted animals. These reactive astrocytes appeared not to impede grafted cells from sending projections into host tissue. Despite the survival of transplanted cells in the traumatically injured brain, hNT cells had no significant effect on posttraumatic neurologic motor function during the acute posttraumatic period. Since hNT cells are transfectable, prolonged survival of these transplanted cells in the posttraumatic milieu suggests that grafted hNT cells may be a suitable means for delivery of therapeutic, exogenous proteins into the CNS for treatment of traumatic brain injury. PMID- 10369561 TI - Immunostimulated glial cells potentiate glucose deprivation-induced death of cultured rat cerebellar granule cells. AB - The present study investigates whether immunostimulated glial expression of inducible nitric oxide synthase influences the glucose deprivation-induced death of rat cerebellar granule cells (CGC). CGC/glia cocultures were immunostimulated by interferon-gamma (200 U/ml) and lipopolysaccharides (1 microg/ml) and 2 days later were challenged by glucose deprivation. Neurotoxicity was assessed by measuring the release of lactate dehydrogenase. Neither a 2-h glucose deprivation nor a 2-day immunostimulation altered the viability of CGC. A 2-day immunostimulation, however, markedly potentiated the glucose deprivation-induced death of CGC. The increased death of glucose-deprived CGC after immunostimulation was mimicked by the nitric oxide (NO) releasing reagent 3-morpholinosydnonimine (SIN-1) and was partially prevented by the NO synthase (NOS) inhibitor N(G) nitroarginine. The increased death of glucose-deprived CGC either after immunostimulation or by SIN-1 was not altered by various N-methyl-D-aspartate (NMDA) and non-NMDA receptor antagonists. Because superoxide dismutase and catalase, which remove superoxide anion, decreased the augmented death of glucose deprived immunostimulated CGC, the reaction of NO with superoxide to form peroxynitrite appears to be implicated in the potentiated neurotoxicity. Our data indicate that immunostimulated glial cells potentiate the death of glucose deprived neurons in part through the expression of inducible NOS but not through NMDA receptor activation. Potentiation of glucose-deprived CGC death by immunostimulated glial cells may be clinically implicated in the tendency of recurrent ischemic insults to be more severe and fatal than an initial ischemic insult. PMID- 10369562 TI - Craniocerebral trauma induces hemorheological disturbances. AB - Several mechanisms are involved in the development of secondary ischemic brain damage, including microthrombi formation, which is thought to play a prominent role. Ninety-four autopsy cases were macro- and microscopically examined by specific staining for fibrin, 74 of which showed cortical contusion after a craniocerebral trauma. Twenty cases with no neurological pathology were used as controls. Traumatic cases comprised 52 males and 22 females, with a mean age of 48 years; most cases died in the first 48 h. The total number of fibrinous microthrombi in a slice of each hemisphere was determined. The mean number of microthrombi found in contused hemisphere was 152 (37-283), with 88 in the contralateral hemisphere (21-139) as compared to 13 (0-27) in control cases. Differences were statistically significant. Globular microthrombi or "shock bodies" (2-60 micro diameter) were present in five cases. Enhanced presence of microthrombi in contused brain areas, higher incidence in young people, an increase in the amount of microthrombi up to the 9th day after injury and involvement of the contralateral hemisphere free of contusion foci were all demonstrated. Microthrombi would therefore seem to be one of the central secondary events after brain trauma to bear in mind when designing treatment strategies. PMID- 10369563 TI - Agents of the "suis-ide diseases" of swine: Actinobacillus suis, Haemophilus parasuis, and Streptococcus suis. AB - In recent years, Actinobacillus suis, Haemophilus parasuis, and Streptococcus suis have emerged as important pathogens of swine, particularly in high health status herds. Their association with a wide range of serious clinical conditions and has given rise to the moniker "suis-ide diseases." These organisms are early colonizers and, for that reason, are difficult to control by management procedures such as segregated early weaning. Vaccination, serodiagnostic testing, and even serotyping are complicated by the presence of multiple serotypes, cross reactive antigens, and the absence of clear markers for virulence. In this review, we discuss our current understanding of the pathogenesis, epidemiology, and management of the causative agents of the "suis-ide diseases" of swine. PMID- 10369564 TI - Relatedness of Streptococcus canis from canine streptococcal toxic shock syndrome and necrotizing fasciitis. AB - The emergence of streptococcal toxic shock syndrome (STSS) and necrotizing fasciitis (NF) in dogs caused by Streptococcus canis has been reported by our laboratory. Since clonal expansion is thought to be partially responsible for the spread of invasive strains of Streptococcus pyogenes in humans, the relatedness of 15 isolates of S. canis from canine STSS and/or NF was examined using pulsed field gel electrophoresis and biotyping; production of proteases and of a CAMP like reaction were also examined. Only 2 of the 15 STSS and/or NF isolates were clonally related, suggesting that the emergence of canine STSS/NF is not the result of clonal expansion of one or more highly virulent strains of S. canis. All of the isolates produced proteases and demonstrated a CAMP-like reaction, which appear to be additional characteristics of S. canis. PMID- 10369565 TI - Presence of Yersinia enterocolitica in tissues of orally-inoculated pigs and the tonsils and feces of pigs at slaughter. AB - In order to study the early events associated with infection of swine by Yersinia enterocolitica, 42 five-week-old crossbred piglets were inoculated per os with approximately 10(8) Y. enterocolitica O:3. Groups of 5 animals (and one negative control) were euthanized 30 min, 3, 6, 12, 24, 48 and 72 h following the infection. Palatine tonsils, retropharyngeal and mesenteric lymph nodes, esophagus, duodenum, jejunum, ileum (and Peyer's patches), stomach, liver, spleen and feces (from colon) were collected and analyzed for the presence of Y. enterocolitica by standard bacteriological procedures. Natural infections were also analyzed, as a complementary study, by taking one-gram samples of fecal material and tonsils from 291 pig carcasses less than 3 h after slaughter and culturing them for Y. enterocolitica using a cold enrichment technique. Within 30 min, Yersinia enterocolitica O:3 was already present at most sites. The presence of Y. enterocolitica in the liver of 3 out of 10 animals and also in the spleen of 3 out of 10 piglets, within the first 3 h postinfection, but not at later times (with one exception), probably indicated a transient bacteremia accompanying the initial stages of infection. The tonsils were colonized in most animals (13/20) as the bacteria remained present from 12 to 72 h postinfection, while only 4 out of 20 fecal samples were found to be positive over the same period. Up to 10(4) colony-forming units of Y. enterocolitica per gram of tonsil and fecal material were recovered. Finally, among the 291 animals sampled at the abattoir, a total of 79 were found positive, 70 of the tonsils sampled were positive, and bacteria were recovered in 17 fecal samples. It is therefore suggested that palatine tonsils are the most reliable tissue for the indication of an infection/colonization by Y. enterocolitica O:3 in swine and that the removal of this tissue during the slaughter process should be considered in order to minimize the possibility of contamination of meat products. PMID- 10369567 TI - A field evaluation of an indirect immunofluorescent antibody test developed to diagnose plasmacytoid leukemia in chinook salmon (Oncorhynchus tshawytscha). AB - An immunofluorescent antibody test (IFAT) developed for the diagnosis for plasmacytoid leukemia was evaluated against histology under field conditions. Previously published results from a laboratory evaluation indicated that the IFAT had a much higher sensitivity than did histology. One hundred seventy-seven moribund chinook salmon from 3 farms located in British Columbia were sampled. Sensitivity, specificity and their respective quality indices were estimated for the IFAT relative to histology. The IFAT was shown to be unreliable, particularly with respect to sensitivity. Cohen's kappa was also calculated and revealed that the agreement between the 2 tests was no better than random. In contrast to previously published results the IFAT did not perform better than histology in the presence of bacterial kidney disease. The results emphasize the importance of evaluating tests in the field conditions in which they are to be used. The possible reasons for the shortcomings of the IFAT are discussed. PMID- 10369566 TI - Identification of Mycobacterium bovis in bovine clinical samples by PCR species specific primers. AB - Tuberculosis, caused by Mycobacterium bovis is emerging as the most important disease affecting cattle. Furthermore, it results in a major public health problem when transmitted to humans. Due to its difficult and non-specific diagnosis, M. bovis has been declared to be one of the etiologic agents causing significant economic loss in the cattle industry. Our group evaluated a more rapid and specific method, based on a new polymerase chain reaction species specific primers, which amplifies a 470-base pair fragment of the M. bovis genome. A total of 275 milk-producing cows were studied by intradermal tuberculin test (ITT) which gave 184 positive and 91 negative cases. From them, 50 animals were taken from a cattle ranch free of tuberculosis. Three different samples were collected from each animal (blood, nasal mucus, and milk). Positive results were obtained from 26 animals by PCR (11.4%), 1 by bacteriological culturing (0.4%) and 1 by bacilloscopy (0.4%). This finding suggests, as in previous reports, that ITT, normally used for bovine tuberculosis detection, has the inconvenience of having a broad range of specificity and sensitivity, and the PCR technique is a more specific and sensitive test to detect infection associated with M. bovis. Therefore, we propose this PCR assay as a useful tool in the epidemiological characterization of infected animals in areas considered to be at high risk of transmission. PMID- 10369568 TI - Bovine polymorphonuclear neutrophil-mediated phagocytosis and an immunoglobulin G2 protease produced by Porphyromonas levii. AB - Acute interdigital phlegmon (AIP) is a commonly occurring anaerobic bacterial infection in cattle. This study examined in vitro the interaction of bovine polymorphonuclear granulocytic neutrophils (PMN) from blood with bacterial species involved in AIP. Polymorphonuclear neutrophils were purified from whole bovine blood, exposed to one of the three putative etiologic agents of AIP and comparatively assessed for phagocytosis using light microscopy. Fusobacterium necrophorum and Prevotella intermedia were effectively phagocytosed by PMN, but Porphyromonas levii was phagocytosed significantly less effectively by PMN. The effect of high titre anti-P. levii bovine serum on antibody-mediated phagocytosis by PMN was also evaluated. High titre serum increased the efficiency of phagocytosis of P. levii by bovine PMN. This was independent of heat labile complement factors. Antibodies specific for P. levii were assessed for protease activity capable of cleaving bovine immunoglobulins (IgG, IgG1, IgG2, and IgM). Partially purified supernatant from broth cultures of P. levii were incubated with biotinylated immunoglobulins (Igs). Samples were taken from times 0 to 72 h and examined using SDS-PAGE followed by Western blot analysis. Streptavidin alkaline phosphatase and NBT-BCIP were used to visualize the Igs for heavy and light chains as well as lower molecular weight fragments of these glycoproteins. Porphyromonas levii produced an immunoglobulin protease which readily cleaved bovine IgG into fragments, but did not act against IgM. Specifically, the enzyme may be a significant virulence factor as it may act to neutralize the antibodies demonstrated necessary for effective PMN-mediated phagocytosis. PMID- 10369569 TI - Comparison of type I and type II bovine viral diarrhea virus infection in swine. AB - Some isolates of type II bovine viral diarrhea virus (BVDV) are capable of causing severe clinical disease in cattle. Bovine viral diarrhea virus infection has been reported in pigs, but the ability of these more virulent isolates of type II BVDV to induce severe clinical disease in pigs is unknown. It was our objective to compare clinical, virologic, and pathologic findings between type I and type II BVDV infection in pigs. Noninfected control and BVDV-infected 2-month old pigs were used. A noncytopathic type I and a noncytopathic type II BVDV isolate were chosen for evaluation in feeder age swine based upon preliminary in vitro and in vivo experiments. A dose titration study was performed using 4 groups of 4 pigs for each viral isolate. The groups were inoculated intranasally with either sham (control), 10(3), 10(5), or 10(7) TCID50 of virus. The pigs were examined daily and clinical findings were recorded. Antemortem and postmortem samples were collected for virus isolation. Neither the type I nor type II BVDV isolates resulted in clinical signs of disease in pigs. Bovine viral diarrhea virus was isolated from antemortem and postmortem samples from groups of pigs receiving the 10(5) and the 10(7) TCID50 dose of the type I BVDV isolate. In contrast, BVDV was only isolated from postmortem samples in the group of pigs receiving the 10(7) TCID50 dose of the type II BVDV isolate. Type I BVDV was able to establish infection in pigs at lower doses by intranasal instillation than type II BVDV. Infection of pigs with a type II isolate of BVDV known to cause severe disease in calves did not result in clinically apparent disease in pigs. PMID- 10369570 TI - Application of the polymerase chain reaction to detect fowl adenoviruses. AB - The possibility of using the polymerase chain reaction (PCR) for the detection of fowl adenoviruses (FAdV) was tested. The optimal reaction parameters were evaluated and defined for purified genomic DNA of type 8 fowl adenovirus (FAdV 8), and then the same conditions were applied for nucleic acid extracted from infected cells. One hundred picograms of purified viral DNA, or 250 FAdV-8 infected cells, were detected by ethidium bromide staining of the PCR products in agarose gels. The sensitivity was increased to 10 pg purified viral DNA, or 25 infected cells, when the PCR products were hybridized with a specific labeled probe. Several field isolates of FAdV and the CELO virus (FAdV serotype 1) could be amplified by the same primers and conditions, but the size of the amplicons was smaller than that for the FAdV-8 PCR product. Other avian viruses and uninfected cell cultures tested negative. PMID- 10369571 TI - A 15-week experimental exposure of pigs to airborne dust with added endotoxin in a continuous flow exposure chamber. AB - The purpose of this study was to evaluate the effect of longterm exposure to airborne dust and endotoxin on the respiratory system of pigs. A continuous flow exposure chamber was built for the purpose of exposing pigs to selected airborne contaminants. Pigs (n = 6) were exposed to a combination of a very fine corn/soybean meal (40.6 mg/m3) with added lipopolysaccharide (LPS; 12.4 microg/m3) for 8 h/d over 5 d for 15 wk (75 d of exposure). Control pigs (n = 6) were housed in a room with minimal contamination of these airborne contaminants. Surprisingly, dust in the exposure chamber and the control room was highly contaminated with peptidoglycan. Changes in the lung were monitored by collecting bronchoalveolar lavage (BAL) fluid for cytology at 5 different time points throughout the exposure period. Blood samples were collected at the same time for hematology. A non-specific respiratory inflammatory response was found in exposed and control pigs, as suggested by the increased neutrophils in BAL fluid and the small inflammatory areas in the lung tissue. No macroscopic lung lesions were observed in control or exposed pigs. The findings in the control pigs imply that even low dust concentrations and possibly peptidoglycan contamination can induce cellular changes in the BAL fluid and that a true control pig does not exist. In addition, the exposed pigs developed a mild eosinophilia, indicating an allergic response to the airborne contaminants. PMID- 10369572 TI - Study of the effect of total serum protein and albumin concentrations on canine fructosamine concentration. AB - The relationship among serum fructosamine concentration and total serum protein and albumin concentrations were evaluated in healthy and sick dogs (diabetics and dogs with insulinoma were not included). Fructosamine was determined using a commercial colorimetric nitroblue tetrazolium method applied to the Technicon RA 500 (Bayer). Serum fructosamine concentration was not correlated to total protein in normoproteinemic (r = 0.03) and hyperproteinemic dogs (r = 0.29), but there was a high correlation (r = 0.73) in hypoproteinemic dogs. Similar comparison between serum fructosamine and albumin concentrations showed middle correlation (r = 0.49) in normoalbuminemic dogs and high degree of correlation (r = 0.67) in hypoalbuminemic dogs. These results showed the importance of recognizing serum glucose concentration as well as total serum protein and albumin concentrations in the assay of canine serum fructosamine concentration. PMID- 10369573 TI - Plasma myeloperoxidase level and polymorphonuclear leukocyte activation in horses suffering from large intestinal obstruction requiring surgery: preliminary results. AB - Myeloperoxidase (MPO) is a specific enzyme of neutrophil azurophilic granules with a strong oxidative activity. Thanks to a radioimmunoassay of equine myeloperoxidase, the authors have observed a significantly higher plasma level of MPO in horses operated for strangulation obstruction of the large intestine (n = 6) than in horses suffering from a non-strangulating displacement of the large intestine (n = 9). For the 2 groups, 3 phases were distinguished: reception (P1), intensive care (P2) and terminal phase (P3). The mean peak values of MPO for these phases were 121.6 ng/mL (P1), 168.6 ng/mL (P2), and 107.0 ng/mL (P3) for the non-strangulating group, and 242.6 ng/mL (P1); 426.0 ng/mL (P2), and 379.5 ng/mL (P3) for the strangulation group. The variations of the mean peak values of plasma MPO were significantly different between the 2 groups and between the different phases. A significant increase of the least square means of MPO was observed between P1 and P2. A significant decrease of the least square means of the number of circulating leukocytes was observed between P1 and P3. Polymorphonuclear neutrophil activation could play a major role in the pathogenesis of acute abdominal disease and endotoxic shock. PMID- 10369574 TI - Biodegradable alginate microspheres as a delivery system for naked DNA. AB - Sodium alginate is a naturally occurring polysaccharide that can easily be polymerized into a solid matrix to form microspheres. These biodegradable microspheres were used to encapsulate plasmid DNA containing the bacterial beta galactosidase (LacZ) gene under the control of either the cytomegalovirus (CMV) immediate-early promoter or the Rous sarcoma virus (RSV) early promoter. Mice inoculated orally with microspheres containing plasmid DNA expressed LacZ in the intestine, spleen and liver. Inoculation of mice with microspheres containing both the plasmid DNA and bovine adenovirus type 3 (BAd3) resulted in a significant increase in LacZ expression compared to those inoculated with microspheres containing only the plasmid DNA. Our results suggest that adenoviruses are capable of augumenting transgene expression by plasmid DNA both in vitro and in vivo. PMID- 10369575 TI - Experimental inoculation of heifers with bovine adenovirus type 3. AB - Nine 2-year-old heifers having BAd3-neutralizing antibody titers between 1:120 and 1:1080 were individually exposed intranasally to an aerosol of 10(8) pfu of wild type (wt) bovine adenovirus type 3 (BAd3). Four animals were kept as non inoculated controls. The heifers were examined daily for rectal temperature, weight gain/loss, nasal and ocular discharges, and other clinical signs for 10 d post-inoculation. None of the animals showed any sign of clinical disease. Virus excretion was observed in one animal only on Day 3 post-inoculation. All BAd3 inoculated heifers demonstrated a significant (P < 0.005, paired t-test) rise in BAd3-specific serum IgG, IgG1, or IgG2 ELISA titers and virus-neutralizing antibody titers compared to the titers before inoculation. All virus-inoculated animals demonstrated increased levels of BAd3-specific IgA ELISA titers in nasal secretions. These results suggest that in the presence of circulating BAd3 neutralizing antibodies, intranasal inoculation of cattle with wt BAd3 would result in inapparent infection. PMID- 10369576 TI - Effects of ambient temperature and scrotal fleece cover on scrotal and testicular temperatures in rams. AB - The objective was to determine scrotal and testicular temperatures in rams and how they are affected by ambient temperature (10 degrees C vs 25 degrees C) and scrotal fleece (densely fleeced vs shaved). Scrotal surface temperatures (SST) of the caudal aspect of the shaved hemi-scrotum at 10 degrees C vs 25 degrees C were (mean, degrees C) 28.9 and 30.5 (P < 0.03), 28.2 and 29.6 (P < 0.04), and 26.1 and 27.6 (P < 0.06) at the top, middle and bottom of the testis, respectively. Scrotal subcutaneous temperatures (SQT) on the fleeced vs shaved side were 33.5 and 32.0 (P < 0.02), 32.2 and 31.1 (P < 0.06), and 31.7 and 30.8 (P < 0.09) at the top, middle, and bottom at 10 degrees C; they were 33.9 and 32.1 (P < 0.02), 33.1 and 31.9 (P < 0.05), and 32.5 and 32.0 (P < 0.15) at 25 degrees C. Intratesticular temperatures (ITT; measured only at 25 degrees C) on the fleeced vs shaved side were 35.3 and 35.0 (P < 0.5), 35.5 and 35.2 (P < 0.4), and 35.4 and 35.0 (P < 0.3) at the top, middle, and bottom. Temperature gradients (difference from top to bottom) were greatest for SST (2.8 degrees C), moderate for SQT (1.8 to 0.1 degrees C), and not significant for ITT (-0.1 and 0.1 degrees C). The SST was approximately 1.5 degrees C warmer at all 3 locations at 25 degrees C vs 10 degrees C. Increased ambient temperature affected SQT more at the bottom than at the top. Conversely, the difference in SQT between the fleeced and shaved sides was greatest at the top. The difference in ITT (0.3 degrees C warmer on the fleeced vs the shaved side at all locations) was not significant. Therefore, the magnitude of temperature increase associated with ambient temperature or scrotal fleece was affected by both depth and vertical location. PMID- 10369577 TI - Motor-vehicle safety: a 20th century public health achievement. AB - The reduction of the rate of death attributable to motor-vehicle crashes in the United States represents the successful public health response to a great technologic advance of the 20th century-the motorization of America. Six times as many people drive today as in 1925, and the number of motor vehicles in the country has increased 11-fold since then to approximately 215 million. The number of miles traveled in motor vehicles is 10 times higher than in the mid-1920s. Despite this steep increase in motor-vehicle travel, the annual death rate has declined from 18 per 100 million vehicle miles traveled (VMT) in 1925 to 1.7 per 100 million VMT in 1997-a 90% decrease. PMID- 10369578 TI - Update: influenza activity--United States and worldwide, 1998-99 season, and composition of the 1999-2000 influenza vaccine. AB - In collaboration with the World Health Organization (WHO), the WHO international network of collaborating laboratories, and state and local health departments, CDC conducts surveillance to monitor influenza activity and to detect antigenic changes in the circulating strains of influenza viruses. This report summarizes surveillance for influenza in the United States and worldwide during the 1998-99 influenza season and describes the composition of the 1999-2000 influenza vaccine. PMID- 10369579 TI - Varicella-related deaths--Florida, 1998. AB - During 1998, the Florida Department of Health (FDH) reported to CDC six fatal cases of varicella (chickenpox). FDH investigated all death certificates for 1998 with any mention of varicella as a contributory or underlying cause. Eight deaths were identified; two were reclassified as disseminated herpes zoster and six were related to varicella, for an annual varicella death rate of 0.4 deaths per million population. Two deaths occurred in children and four in adults; none had received varicella vaccine. The infection source was identified for three cases; two adults acquired varicella from children in the home, and one child acquired varicella from a classmate. One infection source was known to be unvaccinated; the other two were presumed to be unvaccinated. This report summarizes these varicella deaths and recommends prevention strategies. PMID- 10369580 TI - Possible estuary-associated syndrome. AB - Pfiesteria piscicida (Pp) is an estuarine dinoflagellate that has been associated with fish kill events in estuaries along the eastern seaboard and possibly with human health effects. CDC, in collaboration with other federal, state, and local government agencies and academic institutions, is conducting multistate surveillance, epidemiologic studies, and laboratory research for possible estuary associated syndrome (PEAS), including possible Pp-related human illness. PMID- 10369581 TI - Assessment of meibomian gland function in dry eye using meibometry. AB - OBJECTIVE: To study meibomian gland function in dry eyes using meibometry. METHODS: Forty-two patients with clinically diagnosed dry eye that was reclassified as meibomian gland dysfunction (MGD [n = 12]), aqueous-tear deficiency (AD [n = 10]), MGD combined with AD (n = 2), "incomplete" dry eye (n = 12), and non-dry eye (6 eyes) were compared with 41 healthy control subjects. The following 2 techniques of meibometry were applied: direct meibometry (DM) measuring lipid imprints using the Meibometer, and integrated meibometry (IM) using image-scanning and computer densitometry. Tear film lipid layer thickness was assessed using interference microscopy. RESULTS: Imprints were homogeneous for all subjects except those with MGD. Mean+/-SE readings on results of DM were 127.24+/-24.4 for MGD, 306.4+/-9.2 for AD, 248.6+/-13.2 for incomplete dry eye, and 268.5+/-6.3 for controls, showing lower values in the MGD group relative to all others (P<.001). Results of IM gave similar results (P<.001, P =.01, and P<.001, respectively). Lipid layers appeared lower for the MGD group than others. CONCLUSIONS: Compared with controls, lid lipid levels are reduced in patients with MGD, and increased in women with AD. Lipid layer thickness is increased in women with AD compared with patients with MGD. Both meibometric techniques may be useful for evaluating MGD. Although DM requires special equipment (the Meibometer), it provides a record of immediate diagnostic value. Although IM is less effective than DM, it offers visual documentation of the lipid imprint, which may itself be of diagnostic value, and uses equipment available in many laboratories. PMID- 10369582 TI - Quantification of aqueous flare after phacoemulsification with intraocular lens implantation in eyes with pseudoexfoliation syndrome. AB - BACKGROUND: Impairment of the blood-aqueous barrier is a frequent finding in eyes with pseudoexfoliation syndrome (PEX). OBJECTIVE: To perform noninvasive quantification of aqueous flare using the laser flare-cell meter to analyze blood aqueous barrier breakdown following phacoemulsification with intraocular lens implantation in eyes with and without PEX. METHODS: After other conditions that might account for impairment of the blood-aqueous barrier were excluded, 11 eyes with PEX and 11 eyes with senile cataract without PEX were included in the study. Aqueous flare was quantitatively determined using a laser flare-cell meter preoperatively as well as 1, 3, and 5 days postoperatively. Phacoemulsification with posterior chamber intraocular lens implantation was performed by one surgeon. RESULTS: On the first postoperative day, flare values (calculated as mean+/-SD photon counts per millisecond) in eyes with PEX were higher (42.2+/ 21.3) than in eyes without PEX (30.6+/-15.1) (P>.05). On days 3 and 5, postoperative flare values decreased slowly in eyes with PEX (23.9+/-7.4 and 21.2+/-5.7 photon counts per millisecond, respectively) and were significantly higher than in eyes without PEX (14.8+/-5.4 and 10.5+/-1.4 photon counts per millisecond, respectively) (P<.05). CONCLUSIONS: Breakdown of the blood-aqueous barrier is significantly more extensive in eyes with PEX and may be an important risk factor for early postoperative complications. The altered response to surgery should be considered in eyes with PEX. PMID- 10369583 TI - Adenoma of the iris pigment epithelium: a report of 20 cases: the 1998 Pan American Lecture. AB - BACKGROUND: Adenoma of the iris pigment epithelium (IPE) is an uncommon lesion that can simulate iris or ciliary body melanoma, melanocytoma, and pigment epithelial cyst. OBJECTIVES: To evaluate the clinical and pathological features and prognosis of adenoma of the IPE in patients managed by us and to elucidate the features that help to differentiate this tumor from iris melanoma and other similar conditions. PATIENTS AND METHODS: The medical records of 20 patients with adenoma of the IPE were reviewed, and the clinical and histopathologic features were tabulated. RESULTS: Ten patients were male and 10 were female, with a mean age of 60.0 years (range, 11-85 years). All patients were referred because of suspected iris or ciliary body melanoma. All lesions were solitary and unilateral. Sixteen were located in the peripheral iris; 2, in the midzone; and 2, near the pupillary margin. Clinically, all tumors were abruptly elevated, all but 1 were dark gray to black, and all had a smooth, but sometimes multinodular, surface. The tumors caused thinning or complete effacement of the overlying iris stroma, but they did not directly involve the stroma. They typically blocked light with transillumination. On ultrasound biomicroscopy findings, adenoma of the IPE shows a solid tumor pattern, sometimes with small cystoid spaces. The tumor was managed by local resection in 2 patients and observation in 18, all of whom have been stable, with follow-up ranging from 6 months to 9 years. Histopathologic examination revealed a tumor originating in the IPE consisting of cords of pigment epithelial cells separated by septae of connective tissue. CONCLUSIONS: Adenoma of the IPE usually has characteristic features that should differentiate it from iris melanoma, ciliary body melanoma, iris melanocytoma, and iris cyst. Adenoma of the IPE is a benign tumor that may remain relatively stable for years. PMID- 10369584 TI - Macular hole formation: new data provided by optical coherence tomography. AB - OBJECTIVE: To establish the sequence of events leading from vitreofoveal traction to full-thickness macular hole formation. METHODS: Both eyes of 76 patients with a full-thickness macular hole in at least 1 eye were examined by biomicroscopy and optical coherence tomography. RESULTS: Sixty-one fellow eyes had a normal macula. Optical coherence tomograms showed central detachment of the posterior hyaloid over the posterior pole in 19 cases (31%) and a perifoveal hyaloid detachment not detected on biomicroscopy in 26 cases (42%). In the 4 impending macular holes, optical coherence tomography disclosed various degrees of intrafoveal split or cyst, with adherence of the posterior hyaloid to the foveal center and convex perifoveal detachment. In the 14 stage 2 holes, eccentric opening of the roof of the hole was observed, and in the 24 stage 3 holes, the posterior hyaloid was detached from the entire posterior pole. CONCLUSIONS: In fellow eyes of eyes with macular holes posterior hyaloid detachment begins around the macula, but the hyaloid remains adherent to the foveolar center, indicating the action of anteroposterior forces. This results in an intraretinal split evolving into a cystic space, and then to the disruption of the outer retinal layer and the opening of the foveal floor, thus constituting a full-thickness macular hole. PMID- 10369585 TI - Pattern of early visual field loss in HIV-infected patients. AB - OBJECTIVE: To determine the topographic pattern of visual field loss, if any, and its relationship to the stage of disease in human immunodeficiency virus-positive patients without infectious retinopathy. METHODS: A total of 151 eyes from 81 alert and cooperative patients with human immunodeficiency virus were evaluated with visual field testing. Results were analyzed relative to the associated underlying nerve fiber layer patterns associated with retinal ganglion cell axons as they traverse the retina to the optic nerve. The stage of visual field loss was analyzed relative to the length of survival using Kaplan-Meier survival analysis. RESULTS: No correlation of CD4 cell count with visual field mean defect (r2 = 0.23) or corrected pattern standard deviation (r2 = 0.00) was found. A pattern of visual field loss, consistent with sparing of the papillomacular bundles and associated with damage primarily to the inferior retina external to the posterior pole, was found. Survival analysis indicated a significant difference in time of survival between individuals with normal visual fields and those with a diffuse visual field loss, with a trend to less survival with increasing field loss severity. CONCLUSIONS: These results are consistent with disease at the level of the optic nerve. The relationship of stage of visual field loss to survival has important implications for early detection of field loss and appropriate therapeutic intervention to maintain function and quality of life. PMID- 10369586 TI - Use of the polymerase chain reaction to detect B- and T-cell gene rearrangements in vitreous specimens from patients with intraocular lymphoma. AB - OBJECTIVE: To determine whether the polymerase chain reaction for B- and T-cell gene rearrangements could be applied to vitreous specimens to aid in the diagnosis of intraocular lymphoma. METHODS: Vitreous washing specimens from 4 patients were received in balanced saline solution and centrifuged, and a portion of the pellet was used to make routine cytospins. The remainder was used to make a crude extract of DNA that was amplified for immunoglobulin heavy chain and T cell receptor gamma gene rearrangements and the 14;18 translocation by polymerase chain reaction. RESULTS: One patient had 2 specimens 2 years apart. In each, there was an identical band corresponding to the minor cluster region breakpoint of the bcl-2 oncogene, indicating the presence of a 14;18 translocation. One patient showed an immunoglobulin heavy chain gene rearrangement indicating a B cell lymphoma. Two patients showed rearrangements of the T-cell receptor gamma gene, indicating the presence of a T-cell lymphoma. CONCLUSIONS AND CLINICAL RELEVANCE: Vitreous washing specimens can be used successfully to detect B- and T cell gene rearrangements by polymerase chain reaction. This may be useful to confirm the diagnosis of intraocular large cell lymphoma in cases suggestive of the diagnosis. Prompt handling of the specimens is necessary to prevent degradation of the DNA. PMID- 10369587 TI - External beam radiotherapy in retinoblastoma: tumor control and comparison of 2 techniques. AB - OBJECTIVE: To investigate eye conservation, local control, and complication rates among children with retinoblastoma treated with 2 different external beam radiotherapy (EBR) techniques. METHODS: Fifty-eight eyes in 42 patients received EBR as the primary treatment modality for retinoblastoma (median follow-up, 37 months). The EBR technique was relative lens-sparing (RLS) in 26 eyes and modified lateral beam (MLB) in 32 eyes. Both groups were comparable in Reese Ellsworth retinoblastoma classification. If necessary, patients received focal salvage therapy. RESULTS: At 24 months, eye conservation rates were 88.5% and 89.1% among eyes treated with RLS and MLB, respectively (P = .40); tumor control rates without salvage therapy were 84.6% and 53.3% (P = .02), respectively. Among eyes with Reese-Ellsworth stage IV and V disease, eye conservation rates were 88%+/-8% and 83%+/-9% at 36 months in the RLS and MLB groups, respectively, and local tumor control rates were 81%+/-10% and 51%+/-12%. Percentages of eyes without cataract at 36 months were 83.1% and 63.0%, respectively (P = .40). Among patients observed for at least 18 months, midfacial hypoplasia developed in 38.5% and 29.4%, respectively (P = .70). CONCLUSIONS: The EBR technique was associated with high eye conservation and local control rates. Salvage therapy was performed significantly less frequently in the RLS group compared with the MLB group, and complication rates in both groups were similar. PMID- 10369588 TI - Choroidal melanomas with a collar-button configuration: response pattern after iodine 125 brachytherapy. AB - OBJECTIVE: To describe a distinctive type of postbrachytherapy response pattern among choroidal melanomas with a collar-button configuration. METHODS: Ninety three consecutive eyes with choroidal melanoma treated with iodine I 125 brachytherapy before 1991 were reviewed to identify melanomas with a collar button configuration. Postbrachytherapy response patterns of these melanomas were reviewed. RESULTS: Thirty-four of the 93 eyes contained tumors with a collar button configuration. Sixteen (47%) of the 34 eyes demonstrated a postbrachytherapy response pattern characterized by persistence of the collar button, while the body of the tumor demonstrated shrinkage. During follow-up, the color of the collar-button configuration changed from light brown to dark chocolate (16 [47%] of 34 eyes). Subsequently, irregularly clumped, darkly pigmented debris was observed to slough from the surface of the collar-button configuration into the vitreous cavity (7 [21%] of 34 eyes). The debris was comprised largely of pigment-laden macrophages in 1 eye in which a vitrectomy was performed. In another eye, histopathologic study of spherical clusters of intravitreal brown-colored debris identified malignant melanoma cells. CONCLUSIONS: A distinctive postbrachytherapy regression pattern of melanoma with a collar-button configuration has been identified. The main body of the tumor shrinks, whereas the collar-button configuration persists, appears more prominent, gradually changes to a darker color, and may then shed pigmented debris into the vitreous cavity. This pigmented debris may be composed of pigment laden macrophages and/or melanoma cells. Clinical characteristics of the vitreous debris may help distinguish malignant invasion by melanoma cells from infiltration by nonmalignant debris and macrophages. PMID- 10369589 TI - Immunohistochemical distinction of ocular sebaceous carcinoma from basal cell and squamous cell carcinoma. AB - BACKGROUND: Diagnosis of sebaceous carcinoma of the periorbital region is often delayed. Clinically, this lesion can mimic several inflammatory disorders. Histopathologically, it can mimic either squamous cell or basal cell carcinoma. OBJECTIVE: To identify an immunohistochemical approach to assist in the diagnosis of periorbital sebaceous carcinoma. METHOD: The immunohistochemical profiles of several cases of periorbital sebaceous, basal cell, and squamous cell carcinoma were examined. RESULTS: Although at least focal epithelial membrane antigen (EMA) staining can effectively distinguish sebaceous carcinoma (10 of 11 were positive) from basal cell carcinoma (1 of 16 were positive), most squamous cell carcinomas examined were also focally EMA positive (11 of 14). However, Cam 5.2 reactivity was seen in most sebaceous carcinomas (8 of 11) but no squamous cell carcinomas (0 of 14). In addition, at least focal BRST-1 reactivity was also seen in most sebaceous carcinomas (7 of 11) but no basal cell carcinomas (0 of 16). CONCLUSIONS: Periorbital sebaceous, basal cell, and squamous cell carcinomas have different immunohistochemical staining profiles; a panel of commonly available antibodies, including anti-EMA, BRST-1, and Cam 5.2, may help distinguish these diseases from each other when that distinction cannot be clearly made by light microscopy alone. PMID- 10369590 TI - Use of an orbital epidural catheter to control pain after orbital implant surgery. AB - BACKGROUND: The surgical placement of orbital implants for eviscerations, enucleations, and secondary implantations can cause severe postoperative pain that may not be relieved with high doses of narcotics. We analyzed the effectiveness of a method for postoperative pain control in orbital implant surgery using an orbital epidural pain catheter connected to a patient-controlled analgesia bupivacaine hydrochloride pump. METHODS: One hundred nineteen patients undergoing orbital hydroxyapatite implant surgery received placement of an orbital epidural catheter for the infusion of local anesthetics at the conclusion of their surgery. Patients were asked to gauge their level of comfort into the following 3 categories: total, some, or no pain relief in the first week after surgery. A separate numerical grading scale was used to further quantitate pain. Blood samples were collected in 4 patients to assess the systemic levels of bupivacaine. RESULTS: Most patients (88.2%) responded with total or some pain relief, with only 11.8% suffering severe pain. The mean numerical pain score was 2.8, within a range of 0 (no pain) to 10 (severe pain). The average plasma bupivacaine level in the 4 patients in whom this was measured was 0.38 microg/mL, which is well below the toxic level of 4.0 microg/mL. Furthermore, there were only 5 minor complications caused by the catheters, ie, 1 retrobulbar hemorrhage and 4 catheters that did not work. No permanent problems arose from any of the complications. CONCLUSIONS: The orbital epidural pain catheter is an effective means to achieve postoperative pain control after orbital implant surgery. The simple technique of insertion and management of the catheters was well tolerated in our patient population. PMID- 10369591 TI - Effect of melanin on traumatic hyphema in rabbits. AB - OBJECTIVE: To investigate the role of melanin in influencing the clearance of traumatic hyphema and in the incidence of rebleeds following the hyphemas. METHODS: Hyphemas were induced in 30 eyes of New Zealand white albino rabbits using an Nd:YAG laser. A total of 3.75 mg of synthetic melanin suspended in 0.1 mL of balanced salt solution was introduced into the anterior chambers of 16 animals. A total of 0.1 mL of balanced salt solution was injected into 14 control eyes. Hyphema levels were measured by a masked observer (V.D.B.) daily for 15 days. Pairs of animals were sacrificed at 1, 3, 5, 10, and 15 days and the eyes studied histologically. RESULTS: Hyphemas were consistently produced in all eyes with mean+/-SD levels of 1.44+/-0.22 mm and 1.57+/-0.24 mm in the melanin-treated and control eyes, respectively. The clearance of hyphemas in the melanin-treated eyes was significantly prolonged throughout the study (P<.001). The rate of rebleed in the melanin-treated group was 18.8% and in the control group was 7.1% (P<.01). Histologically, both groups showed variable degrees of blood in the anterior chambers and trabecular meshwork. In addition, the melanin-treated eyes showed free melanin, melanin-laden macrophages, and an inflammatory response in the anterior chamber and trabecular meshwork that was greater than that in the control eyes. Melanin-treated eyes with rebleeds showed organized hemorrhage with neovascularization. CONCLUSIONS: The presence of melanin results in a significantly prolonged course of hyphemas and may influence the rate of rebleeds. Occlusion of the trabecular meshwork with melanin-laden macrophages and inflammation may be the mechanisms responsible for these effects. CLINICAL RELEVANCE: The release of melanin into the anterior chamber during ocular trauma may be partly responsible for the susceptibility of darker-pigmented individuals to more serious complications following a traumatic hyphema. PMID- 10369592 TI - Topical prostaglandin F2alpha treatment reduces collagen types I, III, and IV in the monkey uveoscleral outflow pathway. AB - BACKGROUND: Topical prostaglandin F2alpha isopropyl ester increases uveoscleral outflow in monkeys and humans. OBJECTIVE: To investigate the effects of prostaglandin F2alpha isopropyl ester with topical administration on collagen types I, III, and IV within the anterior segment tissue of monkey eyes. METHODS: Eight eyes of 4 cynomolgus monkeys were evaluated. One eye of each monkey was treated with 2 microg of prostaglandin F2alpha isopropyl ester twice daily for 5 days, and intraocular pressure reduction was confirmed. These eyes were fixed in methacarn, and paraffin sections were immunostained using antibodies to collagen types I, II, or IV. To measure staining intensity, optical density (OD) was determined using 2-dimensional imaging densitometry. Mean OD scores along line segments placed over the ciliary muscle were determined. RESULTS: Mean+/-SD OD scores for collagen types I, III, and IV were less in the ciliary muscle of prostaglandin-treated eyes than in vehicle-treated eyes by 52%+/-7%, 45%+/-6%, and 45%+/-5%, respectively. In the sclera adjacent to the ciliary body, mean OD scores for collagen types I and III were less in prostaglandin-treated eyes, by 43%+/-32% and 45%+/-13%, respectively. The scleral stroma was minimally immunoreactive for collagen type IV. All differences were significant by the paired Student t test (P<.05). CONCLUSIONS: This study shows reduced collagen types I, III, and IV immunoreactivity in the ciliary muscle and adjacent sclera following topical prostaglandin F2alpha isopropyl ester treatment. These reductions may contribute to the increased uveoscleral outflow observed with topical prostaglandin treatment. CLINICAL RELEVANCE: The cellular mechanism by which certain prostaglandins lower intraocular pressure is not known. The present study provides immunohistochemical data demonstrating that intraocular pressure reduction that occurs with topical prostaglandin F2alpha is associated with a reduction of collagens within the uveoscleral outflow pathway. PMID- 10369593 TI - Effect of humidity on posterior lens opacification during fluid-air exchange. AB - OBJECTIVE: To study the relationship of humidity and the rate of lens opacity formation during fluid-air exchange using an animal model. METHODS: Vitrectomy and fluid-air exchange was carried out using 16 eyes of 8 pigmented rabbits. One eye of each rabbit was exposed to dry air and the fellow eye received humidified air using an intraocular air humidifier. In each case, the percent humidity of the intraocular air was measured using an in-line hygrometer. Elapsed time from initial air entry to lens feathering was recorded for each eye, with the surgeon observer unaware of the percent humidity of the air infusion. RESULTS: In each rabbit, use of humidified air resulted in a delay in lens feathering (P<.02), with an overall increase in time to feathering of 80% for humidified air vs room air. CONCLUSIONS: Use of a humidifier during fluid-air exchange prolongs intraoperative lens clarity in the rabbit model, suggesting that humidified air should prolong lens clarity during phakic fluid-air exchange in patients. CLINICAL RELEVANCE: Use of humidified air during vitrectomy and fluid-air exchange may retard the intraoperative loss of lens clarity, promoting better visualization of the posterior segment and enhancing surgical performance. PMID- 10369594 TI - Genetic heterogeneity of dominant optic atrophy, Kjer type: Identification of a second locus on chromosome 18q12.2-12.3. AB - OBJECTIVE: To evaluate a family with autosomal dominant optic atrophy, which has been previously linked to the Kidd blood group. DESIGN: Clinical evaluation with the assessment of visual acuity, color vision, and optic nerve appearance to determine affection status. Linkage analysis using polymorphic DNA markers. RESULTS: Visual acuities ranged from 20/20 to 6/200. Although linkage was excluded for chromosome 3q28-29, markers from chromosome 18 in the vicinity of the Kidd locus were linked to the disorder (D18S34 [maximal lod score (lodmax) of 5.38 at recombination fraction (theta) of 0.14], D18S548 [lod(max)=7.26, theta=0.09], D18S861 [lod(max)= 5.32, theta = 0.07], and D18S479 [lod(max) = 3.28, 0 = 0.12:]). Multipoint linkage analysis demonstrated lod scores of greater than 3 in an approximately 3-centimorgan region flanked by D18S34 and D18S479, using 98% penetrance and a phenocopy rate of 1/50. CONCLUSIONS: Dominant optic atrophy is genetically heterogeneous, with loci assigned to chromosomes 3q28-29 and 18q12.2-12.3. Dominant optic atrophy linked to 18q shows intrafamilial variation similar to that previously reported in families linked to 3q, with visual acuities ranging from normal to legal blindness. The overall distribution of visual acuities appears more favorable with the 18q phenotype. Both phenotypes appear to have a similar rate of visual decline. PMID- 10369595 TI - Iris color as a prognostic factor in ocular melanoma. AB - BACKGROUND: Ocular melanoma may be more prevalent among patients with light irises than those with dark irises. OBJECTIVE: To examine a large clinical series of patients with intraocular melanoma to determine if light irises are associated with increased risk of death from these tumors. METHODS: A total of 1162 patients treated with proton irradiation between 1984 and 1996 were observed through 1997. RESULTS: Iris color in the patients was blue or gray in 48%, green or hazel in 30%, and brown in 23%. Tumors in patients with blue or gray irises were less heavily pigmented (P<.001) and closer to the optic disc and macula (P<.001). Five and 10-year metastasis-related death rates were 0.14 and 0.21, respectively, for those with blue or gray irises and 0.10 and 0.15, respectively, for those with darker irises (P = .02). In a Cox proportional hazards regression controlling for tumor characteristics, patients with blue or gray irises died of metastatic disease at a rate 1.90 times (95% confidence interval, 1.26-2.85) that of patients with brown irises. The rate of metastatic death was not significantly elevated for those with green or hazel irises (relative risk, 1.43; 95% confidence interval, 0.91-2.23). CONCLUSION: Patients with blue or gray irises appear to be at increased risk of metastatic death from choroidal melanoma, independent of other risk factors. PMID- 10369596 TI - Practice patterns in diabetic retinopathy: part 1: analysis of retinopathy follow up. AB - OBJECTIVE: To evaluate ophthalmologists' management of diabetic patients. METHODS: A multiple-choice questionnaire was mailed to all ophthalmologists in New York (1985), Florida (1990), and Massachusetts (1993 and 1996). Questions included practice patterns, methods used in examination, use of photography and fluorescein angiography, indications for laser treatment, and intervals for follow-up of selected conditions. Responses were tabulated and compared between surveys and with the American Academy of Ophthalmology Preferred Practice Pattern. RESULTS: In this first report, we detail follow-up patterns of various grades of retinopathy. Physicians increasingly used duration of diabetes as a criterion in determining the length of follow-up for adults, despite persistent ambiguities for children. There were notable changes over time in nearly all aspects of follow-up for both general ophthalmologists and retina specialists. Retina specialists were less likely to treat proliferative disease, more likely to follow up patients with preproliferative disease sooner, and used longer follow-up intervals for short-duration diabetic patients, whereas a small percentage of general ophthalmologists continued to recommend treatment for background disease. CONCLUSION: There were noticeable trends toward Diabetes 2000 recommendations over time, although there remained many areas where further education appeared warranted. PMID- 10369597 TI - Muller cell cone, an overlooked part of the anatomy of the fovea centralis: hypotheses concerning its role in the pathogenesis of macular hole and foveomacualr retinoschisis. AB - Poorly recognized by anatomists and pathologists is the cone-shaped zone of Muller cells that composes the central and inner part of the fovea centralis. The importance of these cells in the structural integrity of the macula, as a repository for xanthophyll, and in the pathogenesis of macular diseases, particularly regarding idiopathic macular hole and foveomacular schisis, is hypothesized. PMID- 10369598 TI - Ethics and medical patents. PMID- 10369599 TI - Harold Ridley, MA, MD, FRCS: a golden anniversary celebration and a golden age. PMID- 10369600 TI - Lichen simplex chronicus of the eyelid. AB - Lichen simplex chronicus is a common dermatosis that rarely affects the eyelids. We report the clinical and pathologic features in the case of a middle-aged man who had lichen simplex chronicus of both lower eyelids. The clinical features suggested the presence of basal cell carcinoma. PMID- 10369601 TI - Benign lymphoid hyperplasia of the conjunctiva in children. AB - Benign lymphoid hyperplasia of the conjunctiva occurs infrequently in children, and its presentation, clinical course, and appropriate management are not well established. We describe 2 children with nasal conjunctival masses that on pathological examination demonstrated benign lymphoid hyperplasia. Local irradiation of residual tissue was deferred, and the lesions remained stable for 1 year in one case and for 3 1/2 years in the other case. No systemic involvement had occurred. Although the natural history of extranodal lymphoid hyperplasia in children is poorly documented, most cases of nodal lymphoid hyperplasia in children are at very low risk of malignant transformation. Careful observation for local and systemic disease is indicated for ocular adnexal lymphoid hyperplasia in children until a more complete understanding of its natural history is available. PMID- 10369602 TI - Foreign body entrapment in radial keratotomy incisions. PMID- 10369603 TI - Ocular inflammation associated with alendronate therapy. PMID- 10369604 TI - Optic disc neovascularization following severe retinoschisis due to shaken baby syndrome. PMID- 10369605 TI - Morning glory disc anomaly in neurofibromatosis type 2. PMID- 10369606 TI - Scurvy causing bilateral orbital hemorrhage. PMID- 10369607 TI - Acute expansion of an orbital vascular malformation. PMID- 10369608 TI - Innovation and clinical trials. PMID- 10369609 TI - Evaluating new retinal imaging techniques. PMID- 10369611 TI - Is the study of blind patients useful for understanding light perception? PMID- 10369610 TI - Retained nuclear fragment 1 year after uncomplicated phocoemulsification cataract extraction with posterior chamber intraocular lens implant. PMID- 10369612 TI - Fosinopril and hydrochlorothiazide combination versus individual components: lack of a pharmacokinetic interaction. AB - OBJECTIVE: To evaluate the pharmacokinetic interaction and bioequivalence of a combination formulation of the angiotensin-converting enzyme inhibitor fosinopril and the diuretic hydrochlorothiazide (HCTZ). METHODS: In an open-label, balanced, randomized incomplete block, three-way crossover fashion, healthy men received single doses of three of four regimens in one of two independent studies. Regimens for study A (36 subjects): (1) fosinopril 10-mg tablet, (2) HCTZ 12.5-mg tablet, (3) a combination tablet of fosinopril 10 mg plus HCTZ 12.5 mg, or (4) coadministered tablets of fosinopril 10 mg and HCTZ 12.5 mg. Study B (40 subjects) received: (1) fosinopril 20-mg tablet, (2) HCTZ 12.5-mg tablet, (3) a combination tablet of fosinopril 20 mg plus HCTZ 12.5 mg, or (4) coadministered tablets of fosinopril 20 mg and HCTZ 12.5 mg. RESULTS: There was no evidence of any significant effect of HCTZ on the pharmacokinetics of fosinoprilat, based on maximum concentration value, AUC, or cumulative urinary recovery over 24 hours. Fosinoprilat had no clinically important effect on the pharmacokinetics of HCTZ only slightly decreasing its AUC by 14% in study A. Coadministration was well tolerated; no new adverse events were reported with the combination tablet. CONCLUSIONS: Fosinopril and HCTZ in a combination tablet display pharmacokinetic profiles similar to those achieved when either drug is administered alone or when coadministered in separate tablets. When used with HCTZ, the favorable pharmacokinetic feature of fosinopril, dual and compensatory pathways of renal and hepatic elimination, is preserved. PMID- 10369613 TI - A critical evaluation of the methodology of the literature on medication compliance. AB - OBJECTIVE: To develop a simple evaluation tool to assess methodologic rigor of the literature on patient compliance with medications, and to apply the tool to a sample of the literature. METHODS: A computerized search of the MEDLINE database (January 1980-December 1996) was performed. All English-language articles on compliance with medications were identified, using the MeSH terms patient compliance and drug-therapy. A 10% sample was then randomly selected for review. Methodologic rigor was assessed on eight standards: study design, specification of patient sample, power analysis, specification of disease, specification of therapeutic regimen, duration of follow-up, definition of compliance, and compliance measurement. The raw scores of the eight standards were then combined in three summary scores, standardized from 0 to 100: study design, disease related features, and compliance issues. RESULTS: Seventy-two articles from 719 identified were reviewed. The majority of the research articles were descriptive (63.9%), and patients in these studies were selected mainly from a convenience sample (41.7%). Just nine studies were multicenter studies, and three employed power analysis. The compliance definition was replicable in 41.7% of the studies. In 22 articles neither the compliance measure nor the criteria were stated. One quarter of the studies (18) used a nonvalidated measure of compliance. Only two studies reached a score of 6 in the compliance measure, and eight studies used two different measures of compliance simultaneously. The median values in the summary scores were: study design 8.3, disease 42.9, compliance issues 50. CONCLUSIONS: The quality of the compliance research was generally poor. These low scores reflect very important shortcomings in the methodology. Such oversights make it difficult for the reader to critically assess the validity of the conclusions. PMID- 10369614 TI - Positive impact of a follow-up phone call to community pharmacies in a medicaid drug utilization program. AB - OBJECTIVE: To evaluate the impact of including community pharmacists in strategies to alter excessive prescribing of antiulcer medications (AUMs) in a statewide drug utilization review (DUR) program. Mailing educational materials to physicians is a common intervention strategy of retrospective DUR programs. However, pharmacists are typically left out of this process, ignoring a possibly influential healthcare provider. METHODOLOGY: Patients without gastroesophageal reflux disease who received > or = 1.0 normalized therapeutic equivalents (e.g., 1.0 NTE = ranitidine 300 mg/d or omeprazole 20 mg/d) for five of six prior months were included. The pharmacists and physicians of these patients were divided into one of three geographically distinct groups: group 1 physicians received mailed materials only (pharmacists were not contacted); group 2 physicians and pharmacists received mailed materials only; and group 3 physicians and pharmacists received mailed materials, and the pharmacists were contacted by phone. Mean NTE and AUM costs were analyzed six months before and six months following the intervention. RESULTS: One hundred thirty-eight, 329, and 248 patients were included in G1, G2, and G3, respectively. There was a 12.4%, 8.0%, and 14.0% reduction in NTE for G1, G2, and G3, respectively. G1 AUM cost decreased 7.7% ($7710); G2 decreased 6.8% ($14 037); G3 decreased 20.5% ($26722). The total decrease in AUM cost for the entire cohort from before to after the intervention was $48469 (p < 0.05) in the six months following the intervention. CONCLUSIONS: A follow-up phone call to pharmacists in a statewide DUR intervention enhances the effectiveness of DUR interventions under the conditions studied. Enlisting the support of community pharmacists may improve the cost savings of these interventions. PMID- 10369615 TI - A comparison of antihypertensive medication utilization before and after guideline changes using the Department of Defense Prescription Database. AB - OBJECTIVE: To determine changes in the proportion of antihypertensive medication utilization, distributed by medication classes, associated with the Fifth Report of the Joint National Committee (JNC V) guideline changes and the Department of Defense Pharmacoeconomic Center's recommendations to follow JNC V guidelines in presumed newly treated hypertensive patients. DESIGN: A 43-month, longitudinal, retrospective analysis using data from the Department of Defense Uniformed Services Prescription Database. SETTING: Seven outpatient US military sites. PATIENTS: Eligible patients (n = 7277) included those from seven military sites, aged 20-49 years, who were: (1) active-duty members of the US Armed Forces, (2) active-duty members' family members, (3) retired members of the US Armed Forces, and (4) US Armed Forces retired members' family members. MAIN OUTCOME MEASURE: Proportion of antihypertensive medication utilization, distributed by medication class over the 43-month study period. METHODOLOGY: Segmented time series analysis was used for each of the following four medication classes: angiotensin converting enzyme inhibitors, beta-blockers, calcium-channel blockers, and diuretics. RESULTS: Segmented time series analyses revealed no significant differences in utilization of the four medication classes that corresponded to published guidelines outlining initial antihypertensive therapy. CONCLUSIONS: It appeared that JNC V guidelines and recommendations of the Pharmacoeconomic Center to follow JNC V had little effect on the utilization of prescription medication classes studied. PMID- 10369616 TI - Seizure associated with olanzapine. AB - OBJECTIVE: To report a case of seizures in a patient who started receiving olanzapine, and to review seizure risk associated with antipsychotic use. CASE SUMMARY: A 31-year-old African-American woman with multiple psychiatric and medical disorders, including generalized seizure disorder, experienced seizure activity when switched from haloperidol to olanzapine. Olanzapine was discontinued, haloperidol was quickly titrated to the previous dose, and the patient was started on oral phenytoin with no further seizure activity noted. The patient remained seizure free and phenytoin was discontinued without complications. DISCUSSION: Determining causality in this case is complicated by the number of confounding factors that may have contributed to the occurrence of seizures in this patient. These factors include: (1) diagnosis of generalized seizure disorder, (2) diagnosis of organic mental disorder, (3) concurrent pharmacotherapy with medications implicated in lowering the seizure threshold, and (4) abrupt change in pharmacotherapy. The likelihood that a significant drug interaction precipitated seizure activity is doubtful. CONCLUSIONS: Considering all factors related to causality, the likelihood that olanzapine was responsible for precipitating seizure activity in this patient was judged possible. Although premarketing studies have indicated that olanzapine may be associated with minimal seizure liability, this case serves as a reminder that postmarketing surveillance of newly released medications is essential. PMID- 10369617 TI - Lamotrigine overdose presenting as anticonvulsant hypersensitivity syndrome. AB - OBJECTIVE: To describe the laboratory and physical manifestations of lamotrigine toxicity presenting as anticonvulsant hypersensitivity syndrome. CASE SUMMARY: A 49-year-old white man presented to our institution with a two-day history of low grade fever, erythema, and edema involving the periorbital area. Five days earlier, he had been placed on lamotrigine treatment for bipolar disorder. He inadvertently received four daily doses of lamotrigine 2700 mg each. Physical examination was significant for periorbital edema and discrete and confluent blanching red macules and papules involving the face, trunk, and extremities. Laboratory tests revealed leukocytosis, hepatitis, and acute renal failure. With normalization of the laboratory results, the eruptions and edema gradually resolved. DISCUSSION: Lamotrigine toxicity can lead to periorbital edema, rash, and multiorgan system abnormalities. This presentation has clinical and laboratory similarities with anticonvulsant hypersensitivity syndrome, which suggests that at some threshold concentration the amount of lamotrigine may overwhelm the body's ability to metabolize the drug, leading to a similar hypersensitivity reaction. Lamotrigine is a relatively new agent, and this report may provide useful insights on evaluating the clinical toxicology of this agent. CONCLUSIONS: Healthcare providers should be aware that lamotrigine overdose may present with multiorgan involvement, similar to anticonvulsant hypersensitivity syndrome. PMID- 10369618 TI - Amoxicillin/clavulanate-associated hepatic failure with progression to Stevens Johnson syndrome. AB - OBJECTIVE: To describe a patient who developed hepatic failure, Stevens-Johnson syndrome (SJS), and died after receiving amoxicillin/clavulanate therapy. CASE SUMMARY: A 37-year-old white man without significant past medical history received a 10-day course of amoxicillin/clavulanate for treatment of pneumonia. Thirty-two days after starting amoxicillin/clavulanate, he developed jaundice, rash, pruritus, and increasing fatigue. On further evaluation, with the exclusion of toxicity from other drugs or diseases, the time course to development of cholestatic jaundice correlated with the use of amoxicillin/clavulanate. The patient consequently died with progressive hepatic failure, renal failure, and SJS. DISCUSSION: Hepatic injury has been reported with amoxicillin/clavulanate. Signs and symptoms of jaundice and pruritus may appear up to to six weeks after stopping therapy. Most cases of liver injury have been benign and reversible on discontinuation of the amoxicillin/clavulanate. Reported hepatic reactions have been mainly cholestatic, with some mixed cholestatic/hepatocellular liver function test abnormalities. CONCLUSIONS: Clinicians should be aware of amoxicillin/clavulanate as a drug capable of causing hepatitis with eventual systemic dysfunction. While recovery is usually complete following withdrawal of the drug, in patients with rash associated with hepatic dysfunction, renal insufficiency, or other unusual symptoms, earlier consideration of initiating systemic steroids or liver transplantation referral, in hopes of avoiding progressive systemic response, might be worthwhile. PMID- 10369619 TI - The use of oral vasopressors in the management of autonomic dysfunction and orthostatic hypotension. AB - OBJECTIVE: To describe a patient with hypotension secondary to autonomic dysfunction who was successfully treated with oral vasopressors. CASE SUMMARY: A 76-year-old African-American man with a history of cerebrovascular accident with right hemiparesis 30 years prior to admission was admitted from another hospital four days after a new posterior inferior cerebellar artery occlusion and poor distal flow as manifested by weakness and hypotension. This was treated with intravenous fluids and dopamine. The dopamine was weaned and changed to phenylephrine to maintain systolic blood pressure >80 mm Hg. Fludrocortisone 0.3 mg orally once daily was initiated; pressure support garments were used for the management of orthostatic hypotension. Ephedrine 25 mg po tid was added and titrated up to 50 mg p.o. tid. Yohimbine 5.4 mg po every eight hours was added due to continued dependence on phenylephrine to maintain adequate blood pressure. Yohimbine was titrated up to 10.8 mg p.o. tid in an unsuccessful effort to wean the patient from phenylephrine. Fludrocortisone was decreased to 0.1 mg po tid and the phenylephrine was tapered off. The patient developed a pan-sensitive Escherichia coli urinary tract infection that was treated with oral trimethoprim/sulfamethoxazole. Over the subsequent days, an 80% left subclavian stenosis was detected; yohimbine and pressure support garments were discontinued. Subsequently, oral ephedrine was tapered off over two days, and fludrocortisone was tapered to 0.1 mg p.o. bid. The patient was transferred in a stable normotensive condition to an inpatient rehabilitation unit. The fludrocortisone was later discontinued with no further hypotension or orthostatic symptoms. DISCUSSION: In this case, orthostatic hypotension associated with autonomic dysfunction was successfully managed with a combination of intravenous vasopressors and hydration, pressure support garments, oral mineralocorticoids, and oral vasopressors. Oral vasopressors and mineralocorticoids are effective treatment options in the management of the vasopressor-dependent patient. In our patient the adverse effects were tolerable. After continued therapy, the oral vasopressors were withdrawn without a return of orthostatic symptoms. CONCLUSIONS: Orthostatic hypotension due to autonomic dysfunction may be successfully managed with combination oral therapy after initial treatment with intravenous vasopressors as evidenced by the absence of orthostasis. PMID- 10369620 TI - Severe multisystemic hypersensitivity reaction to carbamazepine including dyserythropoietic anemia. AB - OBJECTIVE: To report a case of multisystemic hypersensitivity reaction to carbamazepine. CASE SUMMARY: An 81-year-old white man was admitted to our hospital because of fever, morbilliform pruritic rash, and jaundice. Fifty days before admission he had taken carbamazepine 200 mg p.o. tid because of seizures. During the first few days following admission, a maculopapular rash progressed to generalized erythroderma with subsequent extensive skin exfoliation. After discontinuing carbamazepine the fever disappeared within 72 hours and hepatic function tests returned to normal within four days. Moreover, after admission the hemoglobin values gradually fell to 6.7 g/100 mL. A bone marrow aspirate showed hypercellularity with marked dyserythropoietic abnormalities, and the bone marrow biopsy showed large and diffused infiltration due to the presence of a low-grade small lymphocytic lymphoma. No specific therapy for the lymphoma was undertaken. The biochemical follow-up showed a total improvement of hemoglobin values. Eight months after drug discontinuation, the patient was asymptomatic; peripheral blood cell count and hemoglobin concentrations were persistently normal. DISCUSSION: To the best of our knowledge, this is the first published case report implicating carbamazepine as the cause of anemia associated with bone marrow hypercellularity and dyserythropoietic changes, instead of hypocellularity and reduction of erythroid precursors. An interesting point raised by our observation is the possible relation between carbamazepine intake and actual lymphoproliferative disease. The development of non-Hodgkin's lymphoma following carbamazepine treatment has been reported, with regression after the drug was discontinued. However, in our case, a bone marrow biopsy repeated eight months after drug discontinuation confirmed the diagnosis of low-grade lymphoma. CONCLUSIONS: This case report describes a severe multisystemic reaction, characterized by generalized erythroderma; and renal, hepatic, and bone marrow failure in a patient who started carbamazepine therapy 50 days beforehand. PMID- 10369621 TI - Pseudotumor cerebri secondary to intermediate-dose cytarabine HCl. AB - OBJECTIVE: To describe a case of pseudotumor cerebri associated with the administration of intermediate-dose cytarabine. CASE SUMMARY: An 11-year-old Hispanic boy with acute myeloblastic leukemia developed symptoms of pseudotumor cerebri (headache, diplopia, photophobia, nausea, vomiting) after receiving chemotherapy including cytarabine. The patient improved after a lumbar puncture and treatment with prednisone and acetazolamide, and is now asymptomatic. DISCUSSION: Pseudotumor cerebri is a condition usually associated with obese women of child-bearing age. Case reports in pediatric patients are unusual. Several medications have been implicated in causing pseudotumor cerebri, including antimicrobials (tetracycline, naladixic acid), amiodarone, lithium carbonate, vitamin A and its derivatives, growth hormone, and corticosteroids. Chemotherapy agents reported to cause pseudotumor cerebri include busulfan with cyclophosphamide, and the combination of vinblastine, cisplatin, and bleomycin. Most of the information on medication-induced pseudotumor cerebri is in the form of case reports. Different mechanisms for causing this condition have been offered for individual medications. Most of these explanations involve fluid imbalance or interference with the Na+/K+ adenosine triphosphatase pump. Controlled studies are difficult because this condition is an unpredictable and rare occurrence. Cytarabine has frequently been associated with neurologic toxicities, but few reports of pseudotumor cerebri can be found. CONCLUSIONS: The exact cause of pseudotumor cerebri in this patient is unknown, but cytarabine seems a likely cause. The mechanism by which cytarabine could cause this reaction is unclear. PMID- 10369622 TI - Use of intravenous valproate in three pediatric patients with nonconvulsive or convulsive status epilepticus. AB - OBJECTIVE: To report the pharmacokinetics of intravenous valproate (VPA) in children with generalized convulsive status epilepticus (GCSE) or nonconvulsive status epilepticus (NCSE). To provide loading and maintenance dosing for patients with hepatic induction secondary to concurrent anticonvulsants. CASE SUMMARY: Two patients (10 y, 34 mo) with GCSE refractory to benzodiazepines, phenobarbital, phenytoin, and pentobarbital received intravenous VPA. Apparent volume of distribution (Vd) following a 20 mg/kg loading dose was 0.29 L/kg. Maintenance infusions of 4-6 mg/kg/h produced steady-state total concentrations of 66 mg/L and 92.4 mg/L (unbound concentration 44.6 mg/L). Clearance ranged from 63-66 mL/h/kg. An eight-year-old with NCSE received intravenous VPA (13.4 mg/kg load followed by 9 mg/kg every 8 h). Total and unbound steady-state VPA concentrations were 32.9 mg/L and 21.2 mg/L, respectively. Elimination half-life was eight hours. DISCUSSION: We constructed a pharmacokinetic simulation using VPA parameters from children receiving mono- or polyanticonvulsants. Our Vd and elimination half-life rates were comparable with published pediatric values. Patients on hepatic inducers had clearance rates 2.5 times those of children receiving oral anticonvulsant polytherapy. Unbound fractions (48.3% and 66%) were significantly higher than normal. CONCLUSIONS: A 20 mg/kg loading dose should produce a concentration after the bolus dose of approximately 75 mg/L. Initial infusion should consider hepatic induction (noninduced = 1 mg/kg/h, polyanticonvulsant therapy = 2 mg/kg/h, and high-dose pentobarbital = 4 mg/kg/h). Adjustments should be based on response and serum concentrations. PMID- 10369624 TI - Etidronate and alendronate in the treatment of postmenopausal osteoporosis. AB - OBJECTIVE: To review the clinical trials evaluating the efficacy of etidronate and alendronate in the treatment of established postmenopausal osteoporosis. DATA SOURCE: A MEDLINE search was performed (from 1966 through September 1998) using the search terms bisphosphonates, etidronate, alendronate, and postmenopausal osteoporosis. English-language articles were considered for review. STUDY SELECTION AND DATA EXTRACTION: Prospective, randomized, double-blind, placebo controlled clinical trials using fracture as an end point were selected to review the efficacy of etidronate and alendronate in the treatment of postmenopausal osteoporosis. Results for the outcomes of bone mineral density (BMD) and fracture are summarized. DATA SYNTHESIS: Etidronate and alendronate increase spinal BMD in postmenopausal women with osteoporosis. In one study, etidronate decreased the number of women sustaining new radiographic vertebral fractures over two years, but this effect was lost after three years of treatment. Alendronate reduces the number of radiographic vertebral fractures in postmenopausal women with a low bone mass. In women with preexisting fractures, alendronate decreases the number of patients with radiographic vertebral fractures, clinical (i.e., symptomatic vertebral and nonvertebral) fractures, and hip fractures. A significant reduction in the overall number of nonvertebral fractures has not been demonstrated in clinical trials evaluating either alendronate or etidronate. CONCLUSIONS: No studies have directly compared the efficacy of alendronate and etidronate and the results of long-term clinical trials (i.e., >5 y) have not been published. Based on the results obtained in clinical trials using fracture as an end point, alendronate appears to be the bisphosphonate of choice. Safety profiles and cost should also be considered in the choice of etidronate or alendronate for the treatment of postmenopausal osteoporosis. PMID- 10369623 TI - Onycholysis associated with weekly administration of paclitaxel. AB - OBJECTIVE: To report an unusual reaction associated with weekly administration of paclitaxel. CASE SUMMARIES: Onycholysis was seen in four women with recurrent ovarian cancer being treated with low-dose, weekly paclitaxel. Two of the patients had previously received higher doses of paclitaxel on an every-three week schedule without similar reactions. Onycholysis developed between weeks 10 13 of treatment in three of the patients. In the fourth patient, it developed shortly after initiation of weekly paclitaxel. None of the reactions required dose adjustments or discontinuation of therapy. Direct toxicity to the nail bed or inhibition of angiogenesis are possible mechanisms for this reaction. DISCUSSION: Onycholysis, separation of the nail from the nail bed, is an infrequent adverse effect of drug therapy. Antineoplastic drugs have previously been reported to cause onycholysis, pigmentation, bands, thickening or thinning of the nail bed, and nail shedding. Nail changes with the taxanes, primarily docetaxel, are reported in up to 30-40% of patients. Paclitaxel is not commonly associated with dermatologic reactions, although localized skin reactions and tissue necrosis have been reported. Nail changes, pigmentation or discoloration of the nail bed, occur in 2% of patients receiving paclitaxel. CONCLUSIONS: Onycholysis is an uncommon reaction that may occur in some patients receiving weekly, low-dose paclitaxel therapy. The reaction is not life-threatening and does not warrant discontinuation of therapy. However, clinicians should be aware of the possibility of this effect and be prepared to advise patients who develop signs of nail changes. PMID- 10369625 TI - Once-daily aminoglycosides in the treatment of gram-positive endocarditis. AB - OBJECTIVE: To examine the role of once-daily aminoglycosides (ODA) in the treatment of gram-positive endocarditis. DATA SOURCES: A MEDLINE search was conducted from January 1984 to August 1998, and a Current Contents search was performed from September 1998 to December 1998, using endocarditis or aminoglycoside as key words. In addition, relevant articles were cross-referenced to screen for additional information. DATA EXTRACTION: Data published in English regarding the use of aminoglycosides in endocarditis are cited. Emphasis was placed on animal and human studies, but in vitro studies and review articles are also included. DATA SYNTHESIS: Endocarditis and the pharmacology of aminoglycosides are briefly reviewed. ODA is an alternative to conventional dosing in the treatment of endocarditis. Extensive work in endocarditis has been done recently in animals and humans to add to our understanding. Limited clinical data exist to support the theoretical advantages of increased efficacy, reduced toxicity, and potential cost savings versus traditional synergistic aminoglycoside dosing. Optimal monitoring of ODA remains undefined. CONCLUSIONS: Routine use of ODA for the treatment of endocarditis is not yet advocated. Promising supporting evidence and speculation of success of ODA in gram-positive endocarditis justify well-designed trials to further define its role in therapy. PMID- 10369626 TI - Antiinfectives update: focus on treatment and prevention of viral and associated infections. AB - OBJECTIVE: To review the clinically significant antiinfectives approved by the Food and Drug Administration (FDA) since 1996, with an emphasis on agents used for treatment, prevention, or suppression of infection in immunocompromised individuals. DATA SOURCES: A MEDLINE search covering November 1994 to March 1998 was conducted to identify all antiinfectives (new medications and old medications with new indications) and the pertinent literature for review. The search was updated in August 1998 and supplemented with an FDA listing of approved drugs to enhance completeness. STUDY SELECTION: Clinically relevant studies were selected to highlight specific points about each medication. Preclinical publications were used when sufficient information was not available from clinical trials and this information was needed for clinical practice. CONCLUSIONS: Several new and promising antiretroviral agents (stavudine, lamivudine, saquinavir soft-gel capsules, nelfinavir, efavirenz) have been approved, which may allow more options to control HIV viremia. New options for treatment, prevention, and suppression of infections in immunocompromised individuals include azithromycin, cidofovir, famciclovir, valacyclovir, and itraconazole suspension. Liposomal-based amphotericin products may be associated with less toxicity than conventional amphotericin B; however, superior efficacy has not been proven. PMID- 10369627 TI - Chlamydia pneumoniae and coronary heart disease. AB - OBJECTIVE: To review the potential association between Chlamydia pneumoniae (CP) infection and coronary artery disease (CAD), and to describe possible therapeutic interventions. DATA SOURCES: A MEDLINE search of literature (January 1966-January 1998) pertaining to CP infection associated with heart disease was performed. Additional literature was obtained from review of journals and reference lists of pertinent articles identified through the search. STUDY SELECTION AND DATA EXTRACTION: All articles involving CP and CAD were considered for possible inclusion in this review. Other selected articles involved possible links between infection and the atherosclerotic process, inflammation and inflammatory mediators in the atherosclerotic process, and isolation of CP from human tissue. DATA SYNTHESIS: Numerous reports have suggested an association between chronic CP and CAD. CP has been seroepidemiologically linked to CAD. The organism has also been isolated from atherosclerotic lesions. Two reports in humans and one report in animals have shown that macrolide therapy (azithromycin or roxithromycin) may decrease the risk of adverse cardiovascular events. CONCLUSIONS: Evidence seems to support an association between CP infection and an increased incidence of CAD. Additional and larger seroepidemiologic studies of this association need to be performed to establish a causal relationship between infection and CAD. Determination of the actual role of CP in CAD may decide the role of specific antichlamydial therapy in the management of this condition. PMID- 10369628 TI - Clozapine-associated neuroleptic malignant syndrome: two new cases and a review of the literature. AB - BACKGROUND: Clozapine has recently been found to be associated with neuroleptic malignant syndrome (NMS). Our objective is to determine if clozapine causes NMS, if the presentation of clozapine-induced NMS differs from that of traditional agents, and which set of diagnostic criteria will most readily allow diagnosis of NMS associated with clozapine. METHODS: Two new cases of clozapine-associated NMS are presented, along with previously reported cases from the literature, identified by using a MEDLINE search (1966-August 1998). From all cases, concomitant medications and washout periods were examined (if available) to assess clozapine as the likely cause of NMS. Characteristics of clozapine and traditional antipsychotic-induced NMS were compared. Different diagnostic criteria for NMS were applied to the cases to determine which were more likely to diagnose the syndrome. RESULTS: Clozapine was deemed a highly probable cause of NMS in 14 cases, a medium probability cause in five cases, and a low probability cause in eight cases. The most commonly reported clinical features were tachycardia, mental status changes, and diaphoresis. Fever, rigidity, and elevated creatine kinase were less prominent than in NMS associated with classical neuroleptics. CONCLUSIONS: Clozapine appears to cause NMS, although the presentation may be different than that of traditional antipsychotics. Levenson's original and Addonizio's modified criteria were more likely to diagnose NMS than were other criteria. Clozapine-associated NMS may present with fewer clinical features. Limitations are the lack of detailed information provided by many of the case reports and the use of "modified" diagnostic criteria for retrospective diagnosis. PMID- 10369629 TI - Alopecia due to psychotropic medications. AB - OBJECTIVE: To review the literature and describe the incidence and nature of psychotropic drug-related alopecia to assist clinicians in their therapeutic decisions when this adverse event occurs. DATA SOURCES: A MEDLINE search (December 1966-March 1998), using each drug name and the keywords alopecia or hair loss, was conducted. A database from the Clarke Institute of Psychiatry in Toronto was also searched. Additional English-language articles were identified through the bibliography of the reviewed literature. Certain pharmaceutical companies were also consulted. STUDY SELECTION: All published case reports and review articles were considered for study evaluation. DATA EXTRACTION: When possible, details regarding psychotropic drug therapy, development of alopecia, and clinical outcomes were collected for each case. DATA SYNTHESIS: Hair loss from the scalp, eyebrows, and pubic area was identified as a possible, yet uncommon, adverse effect of most psychotropic medications. There are few effective management options for alopecia; however, resolution was achieved on discontinuation of the offending drug in almost all of the cases reported. Alopecia was perceived to be an undesirable effect by most patients, often resulting in poor compliance and hence therapeutic failures. CONCLUSIONS: Due to the self-limiting nature of mild to moderate hair loss with psychotropic medications, clinicians should consider continuing therapy if there is a good clinical response and the patient agrees with this decision. If severe alopecia occurs, it is recommended to discontinue the medication and pursue therapy with another agent. This is to ensure positive therapeutic outcomes and improved patient compliance. PMID- 10369630 TI - The efficacy of extended-interval dosing of omeprazole in keeping gastroesophageal reflux disease patients symptom free. AB - The potential economic advantage of alternate-day therapy for GERD maintenance must be weighed against the potential cost of failure before it can be widely instituted. The studies presented have helped develop a clinical picture of the patients who may benefit from alternate-day therapy without risk of complications or potential increases in management costs. Bank et al., reporting on a group of patients, found that patients with Grade II-IV disease had a 61% success rate at two to eight years. Bank defined success as both maintenance of endoscopic healing and symptom control. Ladas et al. found a 66.7% success rate defined as clinical and endoscopic remission in Grades II-III disease. Kurucar et al. monitored symptom control and esophageal complications in his patients and found the regimen to have a 26% success rate in Grades III-IV disease. Lind et al. found that 83% of patients could remain symptom free with on-demand therapy if they were endoscopy-negative at baseline. The results of the Mantides et al. study are important because they imply that alternate-day omeprazole therapy may be more effective than alternatives for step-down treatment, such as ranitidine or cisapride. Furthermore, patients can be educated to increase their frequency of use if symptoms should arise. Not only does this give the patient a sense of self-empowerment over his or her disease state, but it avoids the cost of switching to a PPI due to failure with an H2RA or a motility agent. Alternate-day use of omeprazole should be attempted only during the maintenance phase of GERD therapy. Patients requiring >20 mg/d to achieve healing appeared to be poor candidates for alternate-day omeprazole maintenance therapy. Based on available studies, it would seem that patients with Grades 0-II GERD would benefit most from alternate-day therapy. A role for alternate-day therapy in Grades III-IV is apparent from the results presented but requires greater caution in view of the differing success rates (26-61%) in various studies. With Grades II-III esophagitis, a mean 24-hour gastric pH >6 and a gastric pH <4 less than 10% of the time during the initial healing phase with omeprazole 20 mg/d appeared to be associated with success on alternate-day therapy. Evidence that all marketed PPIs have similar success is not available and should not be extrapolated from the data presented. Evidence that downward dosage adjustments of PPIs versus extending dosage intervals are effective in the maintenance of GERD should be recognized. Lansoprazole has been approved for treating erosive esophagitis at 30 mg/d, with the maintenance dose established at 15 mg/d. Studies showing that lansoprazole 15 mg/d is more effective than alternate-day therapy with lansoprazole 30 mg exist, although similar studies with omeprazole have not been performed. The abstracts describing the use of alternate-day omeprazole accounted for all enrollees and included endoscopic grading or pH monitoring to document disease severity at baseline. Most also included these same objective measures as end points in combination with symptom control. This strengthens the data since the positive predictive value of typical symptoms is variable. However, there are also several significant limitations. Abstracts provide only limited information on methods. All studies other than Lind et al. lacked randomization. This study was also the only one that blinded patients to their treatment. Sample sizes for the majority of the trials were quite small. Statistical analyses were not performed on any of the trial results with the exception of the trial by Lind et al. In light of the lack of evidence of statistical significance as well as study design flaws, conclusions should be drawn cautiously. Larger well-designed trials looking at both the efficacy and cost-effectiveness of alternate day omeprazole are required before a definitive recommendation can be made. PMID- 10369632 TI - Neuroleptic malignant syndrome associated with metoclopramide. PMID- 10369631 TI - Omeprazole and vitamin B12 deficiency. AB - The mainstay for cobalamin deficiency is correction of the underlying disorder and replacement therapy. Because the defect is often one of absorption, parenteral or intranasal routes are recommended. In most cases, replacement therapy is all that is needed. The vitamin preparation most commonly used is cyanocobalamin (also called vitamin B12), which has no known physiologic role but instead is converted to a biologically active form before it can be used by tissues. The studies reviewed in this article clearly show that omeprazole therapy will decrease the absorption of vitamin B12 by preventing its cleavage from dietary proteins. However, these data are insufficient to infer that clinically significant deficiency will occur over time. In fact, some of the studies suggest that the simple addition of juices or other acidic drinks into the diet may dramatically increase cobalamin absorption. Clearly, well-designed clinical trials are needed to evaluate this theory over an extended follow-up period to determine the clinical significance of omeprazole-associated vitamin B12 deficiency and possibly identify patients at risk for deficiency. In conclusion, the possibility of dietary vitamin B12 malabsorption should be considered in patients receiving chronic omeprazole treatment and presenting with signs and symptoms of deficiency. All healthcare workers should be made aware of the potential clinical complications of omeprazole-associated vitamin B12 deficiency since it may go unrecognized and is easily corrected. This is particularly relevant for elderly patients with poor dietary intake of vitamin B12, impaired vitamin B12 stores, and certain gastrointestinal disorders. PMID- 10369633 TI - Antiretroviral therapy increases serum concentrations of amiodarone. PMID- 10369634 TI - Acral erythema associated with high-dose methotrexate infusion. PMID- 10369635 TI - Utilization and monitoring of aminoglycosides in oncology patients at a Hong Kong government hospital. PMID- 10369636 TI - Current status of cricopharyngeal myotomy for cervical esophageal dysphagia. AB - OBJECTIVE: We have reviewed our experience with cricopharyngeal myotomy for a variety of conditions causing cervical esophageal dysphagia to clarify its indications and results as well as to determine what, if any, ancillary procedures are indicated. METHODS: Eighty-three patients underwent cricopharyngeal myotomy between January 1970 and January 1995, 54 of whom had a pharyngoesophageal diverticulum. The remainder suffered from a variety of motor disorders of the upper esophageal sphincter. Clinical follow-up evaluation was obtained in 71 of the 83 patients (86%). RESULTS: Good or excellent results were obtained in 87% of the patients with pharyngoesophageal diverticula, 100% after myotomy plus diverticulectomy, 87% after myotomy plus diverticulopexy and 67% after myotomy alone. Of patients with hypertensive upper esophageal sphincter, 100% had good or excellent results, whereas only 60% with nonspecific esophageal motor disorders were so evaluated. None of the patients with bulbar palsy or miscellaneous conditions had good or excellent results. CONCLUSIONS: We recommend cricopharyngeal myotomy for all patients with a pharyngoesophageal diverticulum coupled with diverticulopexy for the majority, reserving diverticulectomy for those with recurrent pouches or extremely large pouches (6-8 cm in diameter). Good or excellent results can be expected after myotomy in patients with a hypertensive upper esophageal sphincter. Myotomy is rarely indicated for patients with dysphagia secondary to bulbar palsy. The role of cricopharyngeal myotomy for patients with non-specific esophageal motor disorders remains controversial. PMID- 10369637 TI - Tracheal resection and reconstruction: a review of 82 patients. AB - OBJECTIVE: To assess early and long term (>5 years) results of tracheal resection and reconstruction. PATIENTS AND METHODS: Eighty-two patients amongst 144 with a variety of tracheal lesions, undergoing resection and reconstruction referred to a single surgeon. A retrospective study, the patients were grouped into: Group 1 neoplastic (n = 55) subdivided into: primary tracheal malignancy (PTM, 16), secondary tracheal malignant (STM, 38), benign tracheal neoplasia (BTN, 1); 21 patients in this group had tracheal patch grafts made of Marlex mesh and pericardium; six had a bifurcation resection. Group 2 consisted of non-neoplastic (27) sub-divided into: post-intubation injuries (PII, 24) and traumatic or congenital fistula of the trachea (CTL, 3); 23 patients in this group had circumferential, and the remaining four had partial circumferential, excision of the trachea followed by reconstruction. One patient in this group had tracheal patch graft. Hospital mortality/morbidity, relief of symptoms, recurrence of lesions and long-term survival are considered. RESULTS: Group 1: Five patients (9%) died in hospital; 12 patients, four (two with patch graft) in the PTM, seven (three with patch graft) in the STM group and one in the BTT group survived between 7 and 22 years, one patient is undergoing endoscopic laser. Group 2: There was one death 2 months after surgery. Two patients had recurrence of stricture, one requiring T tube stent, the other endoscopic laser therapy; 24 patients (one with patch graft) remain well between 7 and 22 years. CONCLUSION: Tracheal resection and reconstruction is attended by good functional results. The long-term outcome for malignant disease relates to the histology and tumour staging. Patch grafting using a composite prosthesis of Marlex mesh and pericardium has a chance of long-lasting success when used in selected patients. PMID- 10369638 TI - Is preoperative percutaneous fine-needle aspiration cytology of intrathoracic lesions advisable in resectable patients? AB - OBJECTIVE: The study addresses the clinical significance of percutaneous fine needle aspiration (FNA) cytology in patients with intrathoracic lesions. METHODS: The diagnoses based on cytology in 101 patients (73 male, 28 female; age 21-78 years) with intrathoracic lesions were compared with a definitive histological diagnosis obtained by thoracotomy. Sixty-one lesions were localized in the right and 31 in the left lung, 5 bilaterally and 4 paramediastinally (maximum diameters: 0.8-12 cm; median: 3.5 cm). RESULTS: Upon FNA, 69 cases were graded malignant and 17 benign. In the remaining 15 cases the pathologists felt unable to define clearly the cell type or the biological properties, though the material was found representative. Histology yielded 80 malignant and 21 benign lesions, consistent with the cytological diagnosis in 70 cases. In 60 patients accordance between the cellular subtypes suspected after FNA and those found histologically was present. A significantly higher rate of correct FNA diagnoses was made in malignant lesions (chi-square test: p<0.05). The overall diagnostic accuracy of FNA was 0.77, the sensitivity 0.79 and the specificity 0.91 From the surgical point of view, nine resectable lung cancers, three metastases, three other malignancies and three tuberculomas would have been missed by relying on the FNA diagnoses. Eighteen pneumothoraces (nine requiring suction drainage) occurred after FNA. CONCLUSION: The indication for FNA in otherwise resectable patients should be made carefully, keeping in mind the rate of diagnostic errors and of complications, as well as the possibility for diagnostic VATS of peripheral lesions. PMID- 10369639 TI - Follow-up of patients after resection for bronchogenic carcinoma. AB - OBJECTIVE: To investigate how the members of the European Association for Cardio Thoracic Surgery (EACTS) follow up their patients after pulmonary resection for bronchogenic carcinoma. METHODS: A questionnaire was sent to 317 EACTS members (thoracic and cardiothoracic surgeons as well as surgeons of unknown field of clinical practice). We eventually received completed questionnaires from 101 (31.9%) surgeons, who were classified into "thoracic" and "others". Their answers were analysed by the chi-square test. RESULTS: One out of four EACTS members does not follow up his/her patients, while the remainder follow them up with or without the collaboration of a clinical oncologist, a pneumonologist or a family physician. Among the surgeons who follow up their patients, only one out of two does so throughout the patient's remaining life. The frequency of the routine follow-up visits as well as the type and frequency of the examinations used vary significantly among the members of the Association, but generally the frequency of visits tends to decrease with time. Although 89.8% of surgeons believe that a well scheduled follow-up is beneficial to the patient, only 67% think that such a follow-up is cost-effective. CONCLUSIONS: A great diversity was observed in the way patients operated on for lung cancer are followed up by the EACTS members. The differences were more evident between "thoracic" and "other" surgeons. However, hard data showing the effect of these differences on patients' long-term survival are not available and prospective cooperative studies on this subject are required. Taking into account that these patients are, for the rest of their lives, at high risk of development of a metachronous primary bronchogenic carcinoma or other potentially curable malignancies, we believe that a life-long follow-up is mandatory. PMID- 10369640 TI - Clinical experience with minimally invasive reoperative coronary bypass surgery. AB - OBJECTIVE: To minimize the risk of standard and reoperative coronary artery bypass, we developed a minimally invasive approach. In this study we have evaluated the effectiveness of this technique. METHOD: Between April 1994 and September 1995, 12 men and 6 women, aged 55-84 years (mean, 69 years) with chronic stable angina (4) and recent post-myocardial infarction unstable angina (14), with left ventricular ejection fractions ranging 17-60% (mean 37%), underwent reoperative coronary artery bypass grafting using 7-cm mini-left and right anterior thoracotomy and subxiphoid incisions. Coronary artery anastomoses were carried out on beating hearts with local coronary occlusion. Ischemic preconditioning, beta and calcium channel blockers and the maintenance of mean arterial pressure at 75-80 mm Hg, were used as adjuncts for myocardial protection. The internal mammary artery was isolated under direct vision up to the second rib with excision of the fourth costal cartilage. Coronary artery target sites were the left anterior descending in 12, right coronary artery in 4, obtuse marginal in 3, posterior descending in 1 and diagonal branch in 1 patient. Arterial grafts (mammary, right gastroepiploic, radial), either as single or composite grafts, were used liberally. Preoperative risk factors included congestive heart failure (7), chronic renal insufficiency (5), second reoperation (2), third reoperation (1), cerebrovascular disease (5), prior angioplasty (8) and preoperative intra-aortic balloon pumping in two patients. RESULTS: There was no perioperative mortality with minimal morbidity. Twelve patients underwent patency study of the grafts 48-72 h postoperatively. Ten of the twelve grafts were patent; one internal mammary artery graft to the left anterior descending coronary artery (<1.5 mm) early in our series was occluded and one additional left internal mammary graft had a kink several centimeters away from the anastomosis, which was successfully opened by angioplasty. At a mean follow-up interval of 8 months all 16 surviving patients are in functional class I or II and all of them remain free of angina. CONCLUSION: In selected patients reoperative coronary artery bypass grafting can be performed with this minimally invasive approach with a low perioperative morbidity and mortality rate and satisfactory early graft patency rate with good symptomatic improvement. PMID- 10369641 TI - Intraoperative internal mammary artery transit-time flow measurements: comparative evaluation of two surgical pedicle preparation techniques. AB - OBJECTS: Myocardial revascularization is performed preferentially with internal mammary artery grafts. Pedicle preparation and pharmacologic vasodilatory treatment vary greatly. Objective measurements are difficult since peripheral and later coronary vascular resistance and possible competitive flow of the native bypassed coronary artery will influence the results significantly. Our objectives were: (1) measurement of internal mammary artery graft flow with the transit-time flow technique; (2) comparison of two surgical take-down techniques (skeletonizing vs standard pedicle preparation); (3) quantitation of transit-time flow compared to the free pedicle flow and (4) the vasodilatory effect of papaverine on internal mammary artery flow. METHOD: Consecutive elective cases of coronary artery bypass grafting, performed by two surgeons using routinely either skeletonizing of the internal mammary artery (group A, n = 10) or classical pedicle preparation technique (group B, n = 10), were studied prospectively. Anesthesia, cardiopulmonary bypass and operative data were otherwise comparable; likewise, hemodynamic parameters showed no statistical differences between the two groups. Transit-time flow (CardioMed, Medi-Stim, Norway) was measured at the following time points: beginning (1) and end of take-down (2); after papaverine soaking: before (3) and on cardiopulmonary bypass (4); free flow into a beaker (5); after anastomosis; on (6) and off cardiopulmonary bypass (7). RESULTS: Measurement of mean flow showed the following results: (1) severe vasoconstruction of the internal mammary artery was detected in both groups regardless of the preparation technique (occurring earlier in group A); (2) papaverine soaking caused a moderate flow increase (up to 40%); (3) with corresponding cardiopulmonary bypass flow (4.4 vs. 4.1 l/min in group B) a higher free flow in group A was evident (67.7 vs. 50.7 ml/min); (4) after coronary grafting, transit-time flow showed no significant differences between the two groups and (5) using a 3 mm probe, a linear correlation was demonstrated between transit-time flow and simultaneously measured free flow (r = 0.89). CONCLUSION: Intraoperative transit-time flow measurement is a reliable method for assessing internal mammary artery and coronary artery bypass flow; considering the simple technical application, the procedure may be regarded as a valuable instrument of quality control. PMID- 10369642 TI - Evaluation of the long-term effectiveness of extraluminal and intraluminal vasodilators in an in vitro porcine model of arterial graft spasm. AB - OBJECTIVE: Postoperative graft spasm is a concern when arterial conduits are used because there may be insufficient arterial graft flow. Intraoperatively, vasodilators are used to increase flow and prevent spasm, but little is known about their duration of effectiveness. METHODS: To examine this we attached porcine gastroepiploic and internal thoracic arteries (GEA, n = 48; ITA, n = 24, 10-12 cm long) to a computer-controlled perfusion system (constant inflow pressure 80 mm Hg) with a fixed outflow resistance. Norepinephrine (10(-9)-10(-5) M) was incrementally added to the perfusate at baseline (B), then immediately (h+0) and 2 h (h+2) after the vessels were treated with 30 min of extraluminal or intraluminal nitroglycerin, nitroprusside, verapamil or papaverine. RESULTS: At (B), norepinephrine caused a dose-dependent decrease in flow in both the ITAs and GEAs. In the ITAs, at (h+0), both extraluminal and intraluminal papaverine and, to a lesser extent nitroprusside, increased initial flow and decreased graft sensitivity to norepinephrine. At (h+2), only extraluminal papaverine sustained this maximal effect (ED50 for extraluminal papaverine at (B) 2.6 E(-7) vs. (h+2) 1.3 E(-6), P = 0.01). For the GEAs, at (h+0), both extraluminal and intraluminal verapamil, papaverine, nitroprusside and nitroglycerin attenuated flow reduction due to norepinephrine. At (h+2), only extraluminal papaverine, extraluminal verapamil and intraluminal verapamil were effective in preventing norepinephrine induced spasm (ED50 for extraluminal papaverine at (B) 1.0 E(-7) vs. (h+2) 6.4 E( 6) (P = 0.004); extraluminal verapamil at (B) 1.2 E(-7) vs. (h+2) 4.0 E(-6); intraluminal verapamil at (B) 5.8 E(-7) vs. (h+2) 5.7 E(-6), P = 0.005). CONCLUSION: Verapamil-and papaverine-treated arteries have a greater duration of efficacy in resisting spasm than arteries treated with nitroglycerin and nitroprusside. In the ITA, extraluminal administration of papaverine is most efficacious, possibly due to the prolonged exposure afforded by this route of administration. The effects of verapamil and papaverine are more prolonged in the GEA when administered extraluminally, potentially due to absorption in the perivascular fat-pad and subsequent slow release. The results of this study suggest that extraluminally administered verapamil and papaverine appear to be the preferred vasodilators for preventing arterial graft spasm in the postoperative period. This may be especially important when multiple arterial grafts are used. PMID- 10369643 TI - Autologous platelet sequestration in patients undergoing coronary artery bypass grafting. AB - OBJECTIVES: Blood conservation remains an important issue for patients undergoing cardiac surgery with cardiopulmonary bypass. Platelet sequestration (PSQ) is an aggressive autologous blood conservation method, whose effectiveness is still debated. The main objective of the present study was to evaluate whether PSQ reduces postoperative blood transfusion requirements in patients undergoing coronary artery bypass grafting (CABG) and to determine if PSQ is a cost effective blood conservation method. MATERIAL AND METHODS: All adult patients admitted for CABG entered the study. Exclusion criteria were: recent blood transfusion (<7 days), a platelet count of 150x10(3)/microl or less, hematocrit less than 35% and body weight 50 kg or less. The sequestration was aim 20% or more of the total platelet plasma volume. The sequestration protocol was three sequestration cycles performed just prior to surgery. The concentrated platelet portion was reinfused after weaning from the cardiopulmonary bypass. Hundred seven parameters/patients were recorded. Sixty patients entered the study; 30 in the PSQ group and 30 controls (CTR). RESULTS: Patient characteristics, operation data, preoperative hematology and coagulation parameters did not differ between the groups. In the PSQ group a mean of 433+/-34 ml concentrated platelet portion was collected. The mean platelet count in the concentrated platelet portion was 749+/-157x10(3)/microl, resulting in a platelet yield of 28+/-6% (2040%). The average total chest tube blood loss was 423 ml (PSQ) compared to 858 ml (CTR), p<0.001. A greater number of CTR patients required blood transfusion postoperatively (23) compared to PSQ (3), P<0.001, and fluid requirements were also significantly increased in the control group, P<0.001. No statistical differences in hematology and coagulation parameters between the groups were observed. The hospital mortality was low and the incidence of postoperative complications was few and without group differences. Post-extubation gas exchange was better in PSQ patients compared to CTR. CONCLUSIONS: A preoperative PSQ of a minimum 20% of the total platelet plasma volume resulted in significantly lower postoperative blood loss and fluid and blood transfusion requirements compared to controls. Post-extubation gas exchange was also better after PSQ. Only one patient did not tolerate the sequestration. No other adverse effects of the procedure were observed. PMID- 10369644 TI - New approach for replacement of degenerated mitral bioprostheses. AB - OBJECTIVE: The most common indication for reoperation in patients with a mitral bioprosthetic valve is primary tissue failure. Explanation of the bioprosthesis is time-consuming and may be complicated by cardiac rupture at the atrioventricular junction or the posterior left ventricular wall where a strut is imbedded, injury to the circumflex artery and late perivalvular leak. A new approach to avoid these complications by excising only the bioprosthetic tissue and attaching a reversed aortic St. Jude valve to the intact stent has been developed and evaluated. METHODS: We have replaced degenerated mitral bioprostheses with a St. Jude valve in 73 patients during the last 12 years. In 57, including all who had their operation before 1991, explantation was used. The stent was preserved in 16 patients; in the first four we implanted a mitral St. Jude valve (SJM) within the stent, but this only allows a SJM 6-8 mm smaller than the bioprosthesis. We evolved our approach in the last 12 patients to suture a reversed aortic St. Jude valve with extended cuff to the atrial side of the bioprosthetic cuff; this allows the use of a St. Jude valve 2 mm smaller than the bioprosthesis with exact matching of the orifice sizes. The demographic and clinical profiles of the two groups were similar. RESULTS: Operative mortality was 8/57 (14%) in the explantation group and none in the stent-preservation group. Three late perivalvular leaks occurred in the explanation group, and none in the stent-preservation group. Thirteen late deaths occurred in the explanation group, with a 5-year survival rate of 68%, and one late death (cancer) in the stent-preservation group, but the follow-up is significantly shorter. CONCLUSIONS: Leaving the mitral bioprosthetic stent and cuff intact eliminates the need for extensive dissection, thus shortening and simplifying the procedure and diminishing its attendant mortality and morbidity. It offers a safe and logical approach to replacement of a degenerated mitral bioprosthesis with a St. Jude valve of comparable size which projects into the left atrium, rather than a smaller one jammed into the orifice of the bioprosthetic stent. PMID- 10369645 TI - Long-term results of valve replacement using antibiotic-sterilised homografts in the aortic position. AB - OBJECTIVE: Antibiotic-sterilised homograft valves stored at 4 degrees C have been implanted in the subcoronary position in this unit since 1973. This study was undertaken in order to assess the long-term function of these valves. METHODS: All 249 patients undergoing homograft aortic valve replacement (AVR) at the Wessex Cardiothoracic Centre between April 1973 and December 1994 were studied. Homograft valve sizes ranged from 15 mm to 28 mm internal diameter, 202 (81.1%) varying between 18 mm and 22 mm. The mean patient follow-up was 12.4 years with a total follow-up of 3096 patient-years. There were six early deaths (2.4%). RESULTS: On actuarial analysis, survival was 78.5+/-2.7% (1SE) at 10 years, 65.7+/-3.3% at 15 years and 55.0+/-3.9% at 20 years. The freedom from redo AVR was 87.9+/-2.4% at 10 years, 71.7 +/-3.8% at 15 years and 49.7+/-5.6% at 20 years. The freedom from structural degeneration was 85.6+/-2.5% at 10 years, 63.6+/-4.0% at 15 years and 41.9+/-6.4% at 20 years. On multivariate analysis the risk of valve failure was significantly higher in younger patients (P<0.0001) and in those who underwent aortic root tailoring (P = 0.024). The freedom from endocarditis was 98.4+/-0.9% at 10 years, 96.2+/-1.6% at 15 years and 95.1+/-1.9% at 20 years. Of the 249 patients, 218 had an isolated homograft AVR and were not anticoagulated. In this group there were two possible thromboembolic events. CONCLUSION: As well as the established haemodynamic benefits, this study has shown that homograft AVR with antibiotic-sterilised 4 degrees C stored homograft valves implanted in the subcoronary position, offers good long-term results. PMID- 10369646 TI - Elevated serum levels of S-100 after deep hypothermic arrest correlate with duration of circulatory arrest. AB - OBJECTIVE: Cerebral damage is a major problem after reconstructive surgery of the aortic arch and the descending aorta. Current protective strategies, including deep hypothermia and retrograde cerebral perfusion, are used to prolong the tolerated duration of circulatory arrest, and the latter may also decrease the possibility of air/particle embolization. The aim of the current study was to investigate whether the neurochemical marker S-100 is related to the duration of circulatory arrest, when the influence of embolic injury has been minimized by the use of retrograde cerebral perfusion during the last part of circulatory arrest. METHODS: Arterial serum levels of S-100 were followed before, during and after reconstructive surgery of the thoracic aorta during deep hypothermic arrest in ten adults. Retrograde cerebral blood perfusion was used during the latter part of the arrest period in eight of the ten patients. Neurologic status was followed daily. RESULTS: All patients survived the operation. The median (range) duration of cardiopulmonary bypass (CPB) was 184.5 (121-386) min. The median duration of circulatory arrest and retrograde cerebral perfusion was 50 (3-118) min and 16 (0-84) min, respectively. S-100 increased from 0.10 (0.02-0.18) microg/l preoperatively to 2.37 (0.64-10.80) microg/l after CPB (P<0.01), followed by a decrease to 0.79 (0.21-2.64) microg/l on the first postoperative day (P<0.01). The duration of circulatory arrest correlated with S-100 levels after CPB (r(S) = 0.71, P<0.05) and even better with the S-100 levels on the first postoperative day (r(S) = 0.83, P<0.01). However, there was no significant correlation between duration of arrest and duration of CPB. The duration of circulatory arrest without retrograde cerebral perfusion correlated well with S 100 levels on the first postoperative day (r(S) = 0.88, P<0.01), but not significantly with S-100 levels after CPB. CONCLUSIONS: S-100 levels after aortic surgery with deep hypothermic arrest correlate with the duration of circulatory arrest, indicating that the duration of circulatory arrest is damaging to the brain despite the use of deep hypothermia and partial retrograde cerebral perfusion. The highest correlation between S-100 and duration of arrest was seen on the first postoperative day. S-100 appears to perform well under clinical circumstances as a sensitive and discriminative marker for neuronal injury. PMID- 10369647 TI - Taussig-Bing anomaly and arterial switch: aortic arch obstruction does not influence outcome. AB - OBJECTIVE: Aortic arch obstruction is a commonly associated problem in the Taussig-Bing anomaly. Between 1983 and 1995, 28 consecutive patients with Taussig Bing anomaly underwent arterial switch operation with baffling of the left ventricle to neoaorta. Group A: 11/28 had associated aortic arch obstruction. Group B: 17/28 had isolated Taussig-Bing anomaly. We assessed whether the coexistence of subpulmonary ventricular septal defect and aortic arch obstruction pose an incremental risk factor. PATIENTS AND RESULTS: Group A: Mean age and weight were 1.4+/-1.3 months and 3.5+/-0.4 kg. The aortic arch obstruction included: hypoplasia (5/11), interruption (4/11) and discrete coarctation (2/11). Seven patients had a one-stage correction, and four had initial arch repair followed by arterial switch operation. There were no hospital deaths (CL 0-28%). Over a follow-up of 638 patient-months (mean 64+/-39), there have been no late deaths, and all patients are in New York Heart Association class 1. There have been three cases of recurrent aortic arch obstruction (two requiring reoperation, and one requiring balloon dilation). One patient has been reoperated upon for right ventricular outflow tract obstruction. The actuarial survival and freedom from reoperation rates at 6 years were 100% (CL = 66-100%) and 72.9% (CL=38-92%) respectively. Group B: Mean age and weight were 5.9+/-8.4 months and 5+/-2.1 kg. All patients had a one-stage operation. There were two early deaths (11.8%, CL = 1-36%) and one late death over a follow-up of 678 patient-months (mean 52+/-31). All survivors are in New York Heart Association class 1 and there have been no reoperations. The actuarial survival and freedom from reoperation rates at 6 years were 81% (CL = 56-93%) and 100% (CL = 76-100%) respectively. CONCLUSIONS: 1. Aortic arch obstruction has not adversely affected early or late survival (P>.05) or late functional class. 2. Patients with Taussig-Bing anomaly and aortic arch obstruction may have a higher reoperation rate than those with normal arch anatomy. 3. Taussing-Bing anomaly, with or without aortic arch obstruction, can be repaired with arterial switch operation during the neonatal period with good outcome. PMID- 10369648 TI - A comparison of inhaled nitric oxide with intravenous vasodilators in the assessment of pulmonary haemodynamics prior to cardiac transplantation. AB - OBJECTIVE: Elevated pulmonary vascular resistance and transpulmonary gradient are predictors of increased perioperative mortality in patients undergoing orthotopic heart transplantation. Sodium nitroprusside and prostacyclin PGI2 are routinely used to assess the reversibility of pulmonary vascular resistance and transpulmonary gradient in heart transplant candidates, but their use is limited by their systemic vasodilatory effect. The aim of this study was to evaluate the systemic and pulmonary haemodynamic effects of low concentration (10 and 20 parts per million) inhaled nitric oxide in patients with severe heart failure with elevated transpulmonary gradient and pulmonary vascular resistance undergoing assessment for cardiac transplantation, and to compare the haemodynamic effects of inhaled nitric oxide with those of sodium nitroprusside and prostacyclin PGI2. METHOD: In 10 consecutive patients with elevated transpulmonary gradient (16+/-2 mm Hg) and pulmonary vascular resistance (3.6 +/-0.3 Wood units (WU)) nitric oxide (10 and 20 parts per million in 23% inspired oxygen (O2) via a tight fitting facemask) and increasing doses of intravenous sodium nitroprusside and prostacyclin were administered in a random, single-blinded fashion. RESULTS: Inhalation of nitric oxide (10 ppm) reduced the transpulmonary gradient (-7+/-2 mm Hg; P<0.01) and pulmonary vascular resistance (-1.8+/-0.4 WU; P<0.001) but did not affect the systemic vascular resistance (-0.3+/-1 WU) or mean systemic arterial pressure (-1.3 5 mm Hg). Sodium nitroprusside and prostacyclin reduced the transpulmonary gradient (-4.5+/-2 mm Hg; P<0.01 and -3.6+/-2 mm Hg; P<0.05), pulmonary vascular resistance (-1.5+/-0.4 WU; P<0.001 and -1.3+/-0.4 WU; P<0.01), systemic vascular resistance (-7+/-2 WU; P<0.01 and -7.2+/-2 WU; P<0.01) and mean systemic arterial pressure (-15+/-5 mm Hg; P<0.01 and -18+/-4 mm Hg; P<0.01). CONCLUSION: Low-concentration inhaled nitric oxide is as effective as sodium nitroprusside and prostacyclin in reducing transpulmonary gradient and pulmonary vascular resistance, and is highly pulmonary vasoselective. PMID- 10369649 TI - Reduced oxygen tension during cardiopulmonary bypass limits myocardial damage in acute hypoxic immature piglet hearts. AB - OBJECTIVES: Cardiopulmonary bypass (CPB) is usually instituted in a hyperoxic fashion (oxygen tension (pO2) 300-500 mm Hg), which may expose cyanotic infants to potential reoxygenation damage. Oxygen free radicals play an important role in this injury. The rate of production of this highly reactive toxic oxygen species is dependent on the oxygen level during reoxygenation. This study tested the hypothesis that reduction of the oxygen in the bypass prime and in blood cardioplegia (BCP) to normoxic levels can reduce reoxygenation injury and will result in improved contractility. METHODS: We operated on 19 Duroc-Yorkshire piglets (2-3 weeks, 3-5 kg). Five underwent 30 min of BCP arrest during 1 h of CPB without hypoxia (control). Fourteen underwent 120 min of hypoxia (arterial pO2 20-30 mmHg) on ventilator before reoxygenation on CPB. Reflecting the clinical routine procedure, nine of them were reoxygenated on CPB for 5 min with high pO2 (350-450 mm Hg) followed by 30 min of BCP arrest (high pO2) and 25 min of reoxygenation/reperfusion on CPB with high pO2 levels (NoRx). Five others were put on CPB with pO2 reduced to normoxic levels (pO2 100 mm Hg) in CPB and BCP (Rx). Functional and biochemical measurements were made before hypoxia, as well as during and after reoxygenation. RESULTS: In contrast to controls, NoRx resulted in a 40% decrease in antioxidant reserve capacity (P<0.01) at 4 mM t butyl hydroperoxide (t-BHP), a 1212% increase in myocardial conjugated diene production during BCP induction (P<0.0003), a 1000% during reperfusion (P<0.002), a 36.1% and a 37.0% increase in coronary sinus blood conjugated dienes at 35 min (P<0.05) and 60 min (P<0.05) of reoxygenation. These biochemical changes were accompanied by a 79% reduction of left ventricular contractility (P<0.0003). Conversely, Rx led to an improvement in antioxidant reserve capacity (939+/-212 vs. 1342+/-177 nmol/g protein; P<0.003), less conjugated diene production during BCP induction (15.5+/-6.1 vs. 42.1+/-8.8 A233 nm/min per 100 g; P<0.003) and reperfusion (1.8+/-3.9 vs 22.0+/-5.5 A233 nm/min per 100 g; P<0.005), and to a significantly improved post bypass LV contractility (58+/-25 vs. 21+/-5; P<0.0003). CONCLUSIONS: These data document that hypoxemic/reoxygenation injury occurs in acute hypoxic immature piglet hearts when reoxygenated on CPB with hyperoxic pO2 (normal clinical practice). By lowering the antioxidant reserve capacity, hypoxemia seems to render the developing heart susceptible to reoxygenation damage, which occurs with the reintroduction of molecular oxygen, and is associated with free radical production, subsequent lipid peroxida tion, and depressed post bypass LV function. Reduction of pO2 during the initial reoxygenation period and during BCP arrest to normoxic levels resulted in a significant reduction of this oxygen-related damage and in much improved myocardial performance. PMID- 10369650 TI - Localized intraoperative cardiac tamponade. AB - A 65-year-old lady had undergone mitral and aortic valve replacement following an open mitral valvotomy and aortic valve exploration 5 years earlier. At reoperation, following sternotomy, extensive adhesions were encountered and it was decided to perform minimal dissection of the heart. Both the aortic and mitral valves were replaced using 23 mm and 29 mm St. Jude bileaflet valves, respectively. At the end of the procedure it was difficult to wean the patient off bypass as her mean arterial pressure dropped and the heart became dilated. It was found that a tamponade had developed, as a result of bleeding from the vent site in the pulmonary artery, and dissected a plane between the heart and the adherent pericardium. Her condition improved dramatically as the tamponade was released and she came off cardiopulmonary bypass with no inotropic support. PMID- 10369651 TI - Cor triatriatum sinistrum, aortic coarctation and bicuspid aortic stenosis in an adult. AB - Cardiac anomalies are usually diagnosed early in life, which is particularly true for their various combinations. The diagnosis in adulthood is rare. Here we report the case of a young man with an aortic coarctation corrected at the age of 16, however the associated stenotic bicuspid aortic valve and cor triatriatum sinistrum were corrected after Streptococcus viridans endocarditis 7 years later. PMID- 10369652 TI - Total anomalous pulmonary venous connection to the portal and splenic vein associated with unilateral hypoplasia of pulmonary veins. AB - Total anomalous pulmonary venous return (TAPVR) represents a rare congenital anomaly with wide anatomical and physiological variability. We report a case of a newborn with a challenging form of obstructed infracardiac TAPVR, in whom left and right pulmonary veins drained separately into the portal system. The right pulmonary venous sinus connected to the left branch of the portal vein, whereas the left venous sinus connected to the splenic vein. Surgical repair consisted of the creation of a common retrocardiac venous trunk which was anastomosed to the left atrium. The postoperative course was characterized by persisting congestion of the right lung. Two months later, right pulmonary vein hypoplasia was successfully enlarged with autologous pericardium. PMID- 10369653 TI - Intravenous amiodarone versus propafenone for atrial fibrillation and flutter after cardiac surgery: a note of caution. PMID- 10369654 TI - Isolated congenital left ventricular diverticulum. PMID- 10369655 TI - Ask the doctor. I get palpitations virtually every day. My doctor tells me not to worry about them, but I can't help it. Is there anything I can do? PMID- 10369656 TI - Ask the doctor. I have used a medication called Aldomet for many years for my high blood pressure. Recently, I had to go to the hospital, and the young intern said that he had never heard of anyone using this drug and that it was something out of the history books. Should I be on another drug? PMID- 10369657 TI - Eggcellent news. PMID- 10369658 TI - Speedier mammogram results. PMID- 10369659 TI - Gastrulation in Drosophila: the logic and the cellular mechanisms. AB - The egg contains a set of molecules that can be used to trigger cell-shape changes leading to morphogenetic movements. The temporally and spatially controlled activation of these molecules, and hence the choreography of gastrulation movements, is determined by region-specific expression of transcription factors which turn on a set of downstream targets whose products mediate the successive steps of gastrulation. PMID- 10369660 TI - Specific binding of normal prion protein to the scrapie form via a localized domain initiates its conversion to the protease-resistant state. AB - In the transmissible spongiform encephalopathies, normal prion protein (PrP-sen) is converted to a protease-resistant isoform, PrP-res, by an apparent self propagating activity of the latter. Here we describe new, more physiological cell free systems for analyzing the initial binding and subsequent conversion reactions between PrP-sen and PrP-res. These systems allowed the use of antibodies to map the sites of interaction between PrP-sen and PrP-res. Binding of antibodies (alpha219-232) to hamster PrP-sen residues 219-232 inhibited the binding of PrP-sen to PrP-res and the subsequent generation of PK-resistant PrP. However, antibodies to several other parts of PrP-sen did not inhibit. The alpha219-232 epitope itself was not required for PrP-res binding; thus, inhibition by alpha219-232 was likely due to steric blocking of a binding site that is close to, but does not include the epitope in the folded PrP-sen structure. The selectivity of the binding reaction was tested by incubating PrP res with cell lysates or culture supernatants. Only PrP-sen was observed to bind PrP-res. This highly selective binding to PrP-res and the localized nature of the binding site on PrP-sen support the idea that PrP-sen serves as a critical ligand and/or receptor for PrP-res in the course of PrP-res propagation and pathogenesis in vivo. PMID- 10369661 TI - Molecular basis of glutathione synthetase deficiency and a rare gene permutation event. AB - Glutathione synthetase (GS) catalyses the production of glutathione from gamma glutamylcysteine and glycine in an ATP-dependent manner. Malfunctioning of GS results in disorders including metabolic acidosis, 5-oxoprolinuria, neurological dysfunction, haemolytic anaemia and in some cases is probably lethal. Here we report the crystal structure of human GS (hGS) at 2.1 A resolution in complex with ADP, two magnesium ions, a sulfate ion and glutathione. The structure indicates that hGS belongs to the recently identified ATP-grasp superfamily, although it displays no detectable sequence identity with other family members including its bacterial counterpart, Escherichia coli GS. The difficulty in identifying hGS as a member of the family is due in part to a rare gene permutation which has resulted in a circular shift of the conserved secondary structure elements in hGS with respect to the other known ATP-grasp proteins. Nevertheless, it appears likely that the enzyme shares the same general catalytic mechanism as other ligases. The possibility of cyclic permutations provides an insight into the evolution of this family and will probably lead to the identification of new members. Mutations that lead to GS deficiency have been mapped onto the structure, providing a molecular basis for understanding their effects. PMID- 10369662 TI - Transport of the ADP/ATP carrier of mitochondria from the TOM complex to the TIM22.54 complex. AB - Members of the mitochondrial carrier family such as the ADP/ATP carrier (AAC) are composed of three structurally related modules. Here we show that each of the modules contains a mitochondrial import signal recognized by Tim10 and Tim12 in the intermembrane space. The first and the second module are translocated across the outer membrane independently of the membrane potential, DeltaDeltapsipsi, but they are not inserted into the inner membrane. The third module interacts tightly with the TOM complex and thereby prevents complete translocation of the precursor across the outer membrane. At this stage, binding of a TIM9.10 complex confers a topology to the translocation intermediate which reflects the modular structure of the AAC. The precursor is then transferred to the TIM9.10.12 complex, still interacting with the TOM complex. Release of the precursor from the TOM complex and insertion into the inner membrane by the TIM22.54 complex requires a DeltaDeltapsipsi-responsive signal in the third module. PMID- 10369663 TI - The mechanism mediating regenerative intercellular Ca2+ waves in the blowfly salivary gland. AB - Intercellular Ca2+ signaling in intact salivary glands of the blowfly Calliphora erythrocephala was studied by fluorimetric digital imaging combined with microinjection of putative messenger molecules. Iontophoretic injection of D-myo inositol 1,4, 5-trisphosphate (InsP3) into salivary gland cells evoked regenerative intercellular Ca2+ waves that spread through the impaled cell and several rows of surrounding cells. Ca2+ increases induced by microinjection of Ca2+ ions were confined to the injected cells and their nearest neighbors. Depletion of intracellular Ca2+ stores by thapsigargin pre-treatment did not alter the time course of the Ca2+ increase caused by Ca2+ injection. However, activation of Ca2+ release became clearly evident when Ca2+ was injected in the presence of serotonin (5-HT). Under these conditions, injection of Ca2+ triggered intercellular Ca2+ waves that consecutively passed through >10 cells. The phospholipase C inhibitor U73122 blocked 5-HT-induced Ca2+ increases but did not affect InsP3-dependent Ca2+ spiking and intercellular Ca2+ wave propagation. The results demonstrate that propagation of agonist-evoked Ca2+ waves in the blowfly salivary gland requires supra-basal [InsP3] but does not depend on feedback activation of phospholipase C. We conclude that the intra- and intercellular transmission of these Ca2+ waves is mediated by diffusion of Ca2+ and Ca2+ induced Ca2+ release via the InsP3 receptor channel. PMID- 10369664 TI - Constitutively active Pto induces a Prf-dependent hypersensitive response in the absence of avrPto. AB - Resistance in tomato to Pseudomonas syringae pv tomato (avrPto) is conferred by the gene Pto in a gene-for-gene relationship. A hypersensitive disease resistance response (HR) is elicited when Pto and avrPto are expressed experimentally within the same plant cell. The kinase capability of Pto was required for AvrPto dependent HR induction. Systematic mutagenesis of the activation segment of Pto kinase confirmed the homologous P+1 loop as an AvrPto-binding determinant. Specific amino acid substitutions in this region led to constitutive induction of HR upon expression in the plant cell in the absence of AvrPto. Constitutively active Pto mutants required kinase capability for activity, and were unable to interact with proteins previously shown to bind to wild-type Pto. The constitutive gain-of-function phenotype was dependent on a functional Prf gene, demonstrating activation of the cognate disease resistance pathway and precluding a role for Prf upstream of Pto. PMID- 10369665 TI - Protein tyrosine kinases in bacterial pathogens are associated with virulence and production of exopolysaccharide. AB - In eukaryotes, tyrosine protein phosphorylation has been studied extensively, while in bacteria, it is considered rare and is poorly defined. We demonstrate that Escherichia coli possesses a gene, etk, encoding an inner membrane protein that catalyses tyrosine autophosphorylation and phosphorylation of a synthetic co polymer poly(Glu:Tyr). This protein tyrosine kinase (PTK) was termed Ep85 or Etk. All the E.coli strains examined possessed etk; however, only a subset of pathogenic strains expressed it. Etk is homologous to several bacterial proteins including the Ptk protein of Acinetobacter johnsonii, which is the only other known prokaryotic PTK. Other Etk homologues are AmsA of the plant pathogen Erwinia amylovora and Orf6 of the human pathogen Klebsiella pneumoniae. These proteins are involved in the production of exopolysaccharide (EPS) required for virulence. We demonstrated that like Etk, AmsA and probably also Orf6 are PTKs. Taken together, these findings suggest that tyrosine protein phosphorylation in prokaryotes is more common than was appreciated previously, and that Etk and its homologues define a distinct protein family of prokaryotic membrane-associated PTKs involved in EPS production and virulence. These prokaryotic PTKs may serve as a new target for the development of new antibiotics. PMID- 10369666 TI - IpaC induces actin polymerization and filopodia formation during Shigella entry into epithelial cells. AB - Shigella proteins that are targeted to host cells by a type III secretion apparatus are essential for reorganization of the cytoskeleton during cell invasion. We have developed a semi-permeabilized cell assay that tests the effects of bacterial proteins on the actin cytoskeleton. The Shigella IpaC protein was found to induce the formation of filopodial and lamellipodial extensions in these semi-permeabilized cells. Microinjection of IpaC into cells, or cellular expression of IpaC also led to the formation of filopodial structures. Monoclonal antibodies (mAbs) directed against the C-terminus of IpaC inhibited the IpaC-induced extensions, whereas an anti-N-terminal IpaC mAb stimulated extensive lamellae formation. Shigella induced foci of actin polymerization in the permeabilized cells and these were inhibited by anti-C terminal IpaC mAbs. Consistent with a role for IpaC in Shigella-induced cytoskeletal rearrangements during entry, stable transfectants expressing IpaC challenged with Shigella showed increased bacterial internalization. IpaC-induced extensions were inhibited by a dominant-interfering form of Cdc42 or the Cdc42 binding domain of WASP, whereas a dominant-interfering form of Rac resulted in inhibition of lamellae formation. We conclude that IpaC leads to activation of Cdc42 which in turn, causes activation of Rac, both GTPases being required for Shigella entry. PMID- 10369667 TI - The PrlA and PrlG phenotypes are caused by a loosened association among the translocase SecYEG subunits. AB - prlA mutations in the gene encoding the SecY subunit of the membrane domain of the Escherichia coli preprotein translocase confer many phenotypes: enhanced translocation rates, increased affinity for SecA, diminished requirement for functional leader sequences, reduced proton-motive force (PMF) dependence of preprotein translocation and facilitated translocation of preproteins with folded domains. We now report that both prlA and prlG mutations weaken the associations between the SecY, SecE and SecG subunits of the translocase. This loosened association increases the initiation of translocation by facilitating the insertion of SecA with its bound preprotein but reduces the stimulatory effect of the PMF during the initial step of translocation. Furthermore, the originally isolated prlA4 mutant, which possesses a particularly labile SecYEG complex, acquired a secondary mutation that restored the stability while conserving the flexibility of the complex. Combinations of certain prlA and prlG mutations, known to cause synthetic lethality in vivo, dramatically loosen subunit association and lead to complete disassembly of SecYEG. These findings underscore the importance of the loosened SecYEG association for the Prl phenotypes. We propose a model in which each of the PrlA and PrlG phenotypes derive from this enhanced SecYEG conformational flexibility. PMID- 10369668 TI - Complementation of DsbA deficiency with secreted thioredoxin variants reveals the crucial role of an efficient dithiol oxidant for catalyzed protein folding in the bacterial periplasm. AB - The thiol/disulfide oxidoreductase DsbA is the strongest oxidant of the thioredoxin superfamily and is required for efficient disulfide bond formation in the periplasm of Escherichia coli. To determine the importance of the redox potential of the final oxidant in periplasmic protein folding, we have investigated the ability of the most reducing thiol/disulfide oxidoreductase, E.coli thioredoxin, of complementing DsbA deficiency when secreted to the periplasm. In addition, we secreted thioredoxin variants with increased redox potentials as well as the catalytic a-domain of human protein disulfide isomerase (PDI) to the periplasm. While secreted wild-type thioredoxin and the most reducing thioredoxin variant could not replace DsbA, all more oxidizing thioredoxin variants as well as the PDI a-domain could complement DsbA deficiency in a DsbB-dependent manner. There is an excellent agreement between the activity of the secreted thioredoxin variants in vivo and their ability to oxidize polypeptides fast and quantitatively in vitro. We conclude that the redox potential of the direct oxidant of folding proteins and in particular its reactivity towards reduced polypeptides are crucial for efficient oxidative protein folding in the bacterial periplasm. PMID- 10369669 TI - Glycoprotein reglucosylation and nucleotide sugar utilization in the secretory pathway: identification of a nucleoside diphosphatase in the endoplasmic reticulum. AB - UDP is generated in the lumen of the endoplasmic reticulum (ER) as a product of the UDP-glucose-dependent glycoprotein reglucosylation in the calnexin/calreticulin cycle. We describe here the identification, purification and characterization of an ER enzyme that hydrolyzes UDP to UMP. This nucleoside diphosphatase is a ubiquitously expressed, soluble 45 kDa glycoprotein devoid of transmembrane domains and KDEL-related ER localization sequences. It requires divalent cations for activity and hydrolyzes UDP, GDP and IDP but not any other nucleoside di-, mono- or triphosphates, nor thiamine pyrophosphate. By eliminating UDP, which is an inhibitory product of the UDP-Glc:glycoprotein glucosyltransferase, it is likely to promote reglucosylation reactions involved in glycoprotein folding and quality control in the ER. PMID- 10369670 TI - Early requirement for alpha-SNAP and NSF in the secretory cascade in chromaffin cells. AB - NSF and alpha-SNAP have been shown to be required for SNARE complex disassembly and exocytosis. However, the exact requirement for NSF and alpha-SNAP in vesicular traffic through the secretory pathway remains controversial. We performed a study on the kinetics of exocytosis from bovine chromaffin cells using high time resolution capacitance measurement and electrochemical amperometry, combined with flash photolysis of caged Ca2+ as a fast stimulus. alpha-SNAP, a C-terminal mutant of alpha-SNAP, and NEM were assayed for their effects on secretion kinetics. Two kinetically distinct components of catecholamine release can be observed upon fast step-like elevation of [Ca2+]i. One is the exocytotic burst, thought to represent the readily releasable pool of vesicles. Following the exocytotic burst, secretion proceeds slowly at maintained high [Ca2+]i, which may represent vesicle maturation/recruitment, i.e. some priming steps after docking. alpha-SNAP increased the amplitude of both the exocytotic burst and the slow component but did not change their kinetics, which we examined with millisecond time resolution. In addition, NEM only partially inhibited the slow component without altering the exocytotic burst, fusion kinetics and the rate of endocytosis. These results suggest a role for alpha SNAP/NSF in priming granules for release at an early step, but not modifying the fusion of readily releasable granules. PMID- 10369671 TI - Drainin required for membrane fusion of the contractile vacuole in Dictyostelium is the prototype of a protein family also represented in man. AB - The contractile vacuole expels water by forming a channel with the plasma membrane and thus enables cells to survive in a hypo-osmotic environment. Here we characterize drainin, a Dictyostelium protein involved in this process, as the first member of a protein family represented in fission yeast, Caenorhabditis elegans and man. Gene replacement in Dictyostelium shows that drainin acts at a checkpoint of channel formation between the contractile vacuole and the plasma membrane. A green fluorescent protein fusion of drainin localizes specifically to the contractile vacuole and rescues its periodic discharge in drainin-null cells. Drainin is a peripheral membrane protein, requiring a short hydrophobic stretch in its C-terminal region for localization and function. We suggest that drainin acts in a signaling cascade that couples a volume-sensing device in the vacuolar membrane to the membrane fusion machinery. PMID- 10369672 TI - Extracellular links in Kir subunits control the unitary conductance of SUR/Kir6.0 ion channels. AB - Potassium (K+) channels are highly selective for K+ ions but their unitary conductances are quite divergent. Although Kir6.1 and Kir6.2 are highly homologous and both form functional K+ channels with sulfonylurea receptors, their unitary conductances measured with 150 mM extracellular K+ are approximately 35 and 80 pS, respectively. We found that a chain of three amino acid residues N123-V124-R125 of Kir6.1 and S113-I114-H115 of Kir6.2 in the M1-H5 extracellular link and single residues M148 of Kir6.1 and V138 of Kir6.2 in the H5-M2 link accounted for the difference. By using a 3D structure model of Kir6.2, we were able to recognize two independent plausible mechanisms involved in the determination of single channel conductance of the Kir6.0 subunits: (i) steric effects at Kir6.2V138 or Kir6.1M148 in the H5-M2 link influence directly the diffusion of K+ ions; and (ii) structural constraints between Kir6.2S113 or Kir6. 1N123 in the M1-H5 link and Kir6.2R136 or Kir6.1R146 near the H5 region control the conformation of the permeation pathway. These mechanisms represent a novel and possibly general aspect of the control of ion channel permeability. PMID- 10369673 TI - Tolerance to toxic metals by a gene family of phytochelatin synthases from plants and yeast. AB - Phytochelatins play major roles in metal detoxification in plants and fungi. However, genes encoding phytochelatin synthases have not yet been identified. By screening for plant genes mediating metal tolerance we identified a wheat cDNA, TaPCS1, whose expression in Saccharomyces cerevisiae results in a dramatic increase in cadmium tolerance. TaPCS1 encodes a protein of approximately 55 kDa with no similarity to proteins of known function. We identified homologs of this new gene family from Arabidopsis thaliana, Schizosaccharomyces pombe, and interestingly also Caenorhabditis elegans. The Arabidopsis and S.pombe genes were also demonstrated to confer substantial increases in metal tolerance in yeast. PCS-expressing cells accumulate more Cd2+ than controls. PCS expression mediates Cd2+ tolerance even in yeast mutants that are either deficient in vacuolar acidification or impaired in vacuolar biogenesis. PCS-induced metal resistance is lost upon exposure to an inhibitor of glutathione biosynthesis, a process necessary for phytochelatin formation. Schizosaccharomyces pombe cells disrupted in the PCS gene exhibit hypersensitivity to Cd2+ and Cu2+ and are unable to synthesize phytochelatins upon Cd2+ exposure as determined by HPLC analysis. Saccharomyces cerevisiae cells expressing PCS produce phytochelatins. Moreover, the recombinant purified S.pombe PCS protein displays phytochelatin synthase activity. These data demonstrate that PCS genes encode phytochelatin synthases and mediate metal detoxification in eukaryotes. PMID- 10369674 TI - Activation of the erythropoietin receptor by the gp55-P viral envelope protein is determined by a single amino acid in its transmembrane domain. AB - The spleen focus forming virus (SFFV) gp55-P envelope glycoprotein specifically binds to and activates murine erythropoietin receptors (EpoRs) coexpressed in the same cell, triggering proliferation of erythroid progenitors and inducing erythroleukemia. Here we demonstrate specific interactions between the single transmembrane domains of the two proteins that are essential for receptor activation. The human EpoR is not activated by gp55-P but by mutation of a single amino acid, L238, in its transmembrane sequence to its murine counterpart serine, resulting in its ability to be activated. The converse mutation in the murine EpoR (S238L) abolishes activation by gp55-P. Computational searches of interactions between the membrane-spanning segments of murine EpoR and gp55-P provide a possible explanation: the face of the EpoR transmembrane domain containing S238 is predicted to interact specifically with gp55-P but not gp55-A, a variant which is much less effective in activating the murine EpoR. Mutational studies on gp55-P M390, which is predicted to interact with S238, provide additional support for this model. Mutation of M390 to isoleucine, the corresponding residue in gp55-A, abolishes activation, but the gp55-P M390L mutation is fully functional. gp55-P is thought to activate signaling by the EpoR by inducing receptor oligomerization through interactions involving specific transmembrane residues. PMID- 10369675 TI - The C-terminus of the kinase-defective neuregulin receptor ErbB-3 confers mitogenic superiority and dictates endocytic routing. AB - Signaling by the epidermal growth factor (EGF) family and the neuregulin group of ligands is mediated by four ErbB receptor tyrosine kinases, that form homo- and heterodimeric complexes. Paradoxically, the neuregulin receptor ErbB-3 is devoid of catalytic activity, but its heterodimerization with other ErbBs, particularly the ligand-less ErbB-2 oncoprotein of carcinomas, reconstitutes superior mitogenic and transforming activities. To understand the underlying mechanism we constructed a chimeric EGF-receptor (ErbB-1) whose autophosphorylation C-terminal domain was replaced by the corresponding portion of ErbB-3. Consistent with the possibility that this domain recruits a relatively potent signaling pathway(s), the mitogenic signals generated by the recombinant fusion protein were superior to those generated by ErbB-1 homodimers and comparable to the proliferative activity of ErbB-2/ErbB-3 heterodimers. Upon ligand binding, the chimeric receptor recruited an ErbB-3-specific repertoire of signaling proteins, including Shc and the phosphatidylinositol 3-kinase, but excluding the ErbB-1-specific substrate, phospholipase Cgamma1. Unlike ErbB-1, which is destined to lysosomal degradation through a mechanism that includes recruitment of c-Cbl and receptor poly-ubiquitination, the C-terminal tail of ErbB-3 shunted the chimeric protein to the ErbB-3-characteristic recycling pathway. These observations attribute the mitogenic superiority of ErbB-3 to its C-terminal tail and imply that the flanking kinase domain has lost catalytic activity in order to restrain the relatively potent signaling capability of the C-terminus. PMID- 10369676 TI - Pim kinase expression is induced by LTP stimulation and required for the consolidation of enduring LTP. AB - In animals and several cellular models of synaptic plasticity, long-lasting changes in synaptic strength are dependent on gene transcription and translation. Here we demonstrate that Pim-1, a serine/threonine kinase closely related to Pim 2 and Pim-3, is induced in hippocampus in response to stimuli that evoke long term potentiation (LTP). Mice deficient for Pim-1 show normal synaptic transmission and short-term plasticity. However, they fail to consolidate enduring LTP even though Pim-2 and Pim-3 are constitutively expressed in the hippocampus and Pim-3 expression is similarly induced by synaptic activity. Thus, expression of Pim-1 is required for LTP. Its level of expression and, consequently, its capacity to phosphorylate target proteins in dendritic and nuclear compartments of stimulated neurons might be a determining factor for the establishment of long-lasting changes in synaptic strength. PMID- 10369677 TI - Catabolite control of Escherichia coli regulatory protein BglG activity by antagonistically acting phosphorylations. AB - In bacteria various sugars are taken up and concomitantly phosphorylated by sugar specific enzymes II (EII) of the phosphoenolpyruvate:sugar phosphotransferase system (PTS). The phosphoryl groups are donated by the phosphocarrier protein HPr. BglG, the positively acting regulatory protein of the Escherichia coli bgl (beta-glucoside utilization) operon, is known to be negatively regulated by reversible phosphorylation catalyzed by the membrane spanning beta-glucoside specific EIIBgl. Here we present evidence that in addition BglG must be phosphorylated by HPr at a distinct site to gain activity. Our data suggest that this second, shortcut route of phosphorylation is used to monitor the state of the various PTS sugar availabilities in order to hierarchically tune expression of the bgl operon in a physiologically meaningful way. Thus, the PTS may represent a highly integrated signal transduction network in carbon catabolite control. PMID- 10369678 TI - A mosaic analysis in Drosophila fat body cells of the control of antimicrobial peptide genes by the Rel proteins Dorsal and DIF. AB - Expression of the gene encoding the antifungal peptide Drosomycin in Drosophila adults is controlled by the Toll signaling pathway. The Rel proteins Dorsal and DIF (Dorsal-related immunity factor) are possible candidates for the transactivating protein in the Toll pathway that directly regulates the drosomycin gene. We have examined the requirement of Dorsal and DIF for drosomycin expression in larval fat body cells, the predominant immune-responsive tissue, using the yeast site-specific flp/FRT recombination system to generate cell clones homozygous for a deficiency uncovering both the dorsal and the dif genes. Here we show that in the absence of both genes, the immune-inducibility of drosomycin is lost but can be rescued by overexpression of either dorsal or dif under the control of a heat-shock promoter. This result suggests a functional redundancy between both Rel proteins in the control of drosomycin gene expression in the larvae of Drosophila. Interestingly, the gene encoding the antibacterial peptide Diptericin remains fully inducible in the absence of the dorsal and dif genes. Finally, we have used fat body cell clones homozygous for various mutations to show that a linear activation cascade Spaetzle--> Toll-->Cactus- >Dorsal/DIF leads to the induction of the drosomycin gene in larval fat body cells. PMID- 10369679 TI - Net, a negative Ras-switchable TCF, contains a second inhibition domain, the CID, that mediates repression through interactions with CtBP and de-acetylation. AB - Signalling cascades are integrated at the transcriptional level by the interplay between factors such as the ternary complex factors (TCFs) that interact with serum response factor (SRF) and the serum response element (SRE) of the fos promoter. Net is a negative TCF that is switched to a positive regulator by the Ras signal. To understand the mechanisms of repression by Net, we used a yeast two-hybrid screen to identify factors that interact with its inhibitory domain. We isolated mCtBP1, the murine homologue of huCtBP1, a factor implicated in negative regulation of transformation by E1A plus Ras. We show that mCtBP1 interacts strongly with Net both in vitro and in vivo. The CtBP interaction domain of Net, the CID, mediates repression independently of the previously identified negative element, the NID. The CID inhibits by recruiting the co repressor mCtBP1. The CID and mCtBP1 need to use de-acetylase activity for repression, whereas the NID apparently represses by other mechanisms. Finally, we provide evidence that CtBP and de-acetylation repress the c-fos SRE in low serum when it is inactive, but not in high serum when it is active. These results provide insights into the cross-talk between pathways that inhibit and stimulate transformation at the level of Net, a regulator of gene expression. PMID- 10369680 TI - RYBP, a new repressor protein that interacts with components of the mammalian Polycomb complex, and with the transcription factor YY1. AB - The products of the Polycomb group (PcG) of genes are necessary for the maintenance of transcriptional repression of a number of important developmental genes, including the homeotic genes. A two-hybrid screen was used to search for putative new members of the PcG of genes in mammals. We have identified a new Zn finger protein, RYBP, which interacts directly with both Ring1 proteins (Ring1A and Ring1B) and with M33, two mutually interacting sets of proteins of the mammalian Polycomb complex. Ring1 binds RYBP and M33 through the same C-terminal domain, whereas the RYBP-M33 interaction takes place through an M33 domain not involved in Ring1 binding. RYBP also interacts directly with YY1, a transcription factor partially related to the product of the Drosophila pleiohomeotic gene. In addition, we show here that RYBP acts as a transcriptional repressor in transiently transfected cells. Finally, RYBP shows a dynamic expression pattern during embryogenesis which initially overlaps partially that of Ring1A in the central nervous system, and later becomes ubiquitous. Taken together, these data suggest that RYBP may play a relevant role in PcG function in mammals. PMID- 10369681 TI - Aiolos transcription factor controls cell death in T cells by regulating Bcl-2 expression and its cellular localization. AB - We searched for proteins that interact with Ras in interleukin (IL)-2-stimulated or IL-2-deprived cells, and found that the transcription factor Aiolos interacts with Ras. The Ras-Aiolos interaction was confirmed in vitro and in vivo by co immunoprecipitation. Indirect immunofluorescence shows that IL-2 controls the cellular distribution of Aiolos and induces its tyrosine phosphorylation, required for dissociation from Ras. We also identified functional Aiolos-binding sites in the Bcl-2 promoter, which are able to activate the luciferase reporter gene. Mutation of Aiolos-binding sites within the Bcl-2 promoter inhibits transactivation of the reporter gene luciferase, suggesting direct control of Bcl 2 expression by Aiolos. Co-transfection experiments confirm that Aiolos induces Bcl-2 expression and prevents apoptosis in IL-2-deprived cells. We propose a model for the regulation of Bcl-2 expression via Aiolos. PMID- 10369682 TI - Ptx1 regulates SF-1 activity by an interaction that mimics the role of the ligand binding domain. AB - Ptx1 (Pitx1) is a bicoid-related homeobox transcription factor expressed from the onset of pituitary development. It was shown to cooperate with cell-restricted factors, such as Pit1, NeuroD1/PanI and steroidogenic factor 1 (SF-1), to establish a combinatorial code conferring lineage- and promoter-specific gene transcription in the pituitary. Transcriptional synergism between Ptx1 and SF-1 on two SF-1 target genes, pituitary luteinizing hormone beta and Mullerian inhibiting substance (MIS), requires SF-1 binding to DNA and appears to result from direct physical interaction between these two proteins. The interaction between the C-terminus of Ptx1 and the N-terminal half of SF-1 results in transcriptional enhancement that equals the activity of a constitutively active SF-1 mutant and that may mimic the effect of a still unidentified SF-1 ligand. Thus, the unmasking of SF-1 activity by Ptx1 may represent a developmental mechanism to alleviate the need for SF-1 ligand in transcription and, possibly, at critical times during organogenesis. PMID- 10369683 TI - Promoter upstream bent DNA activates the transcription of the Clostridium perfringens phospholipase C gene in a low temperature-dependent manner. AB - The phospholipase C gene (plc) of Clostridium perfringens possesses three phased A-tracts forming bent DNA upstream of the promoter. An in vitro transcription assay involving C.perfringens RNA polymerase (RNAP) showed that the phased A tracts have a stimulatory effect on the plc promoter, and that the effect is proportional to the number of A-tracts, and more prominent at lower temperature. A gel retardation assay and hydroxyl radical footprinting revealed that the phased A-tracts facilitate the formation of the RNAP-plc promoter complex through extension of the contact region. The upstream (UP) element of the Escherichia coli rrnB P1 promoter stimulated the downstream promoter activity temperature independently, differing from the phased A-tracts. When the UP element was placed upstream of the plc promoter, low temperature-dependent stimulation was observed, although this effect was less prominent than that of the phased A-tracts. These results suggest that both the phased A-tracts and UP element cause low temperature-dependent activation of the plc promoter through a similar mechanism, and that the more efficient low temperature-dependent activation by the phased A tracts may be due to an increase in the bending angle at a lower temperature. PMID- 10369684 TI - Sm and Sm-like proteins assemble in two related complexes of deep evolutionary origin. AB - A group of seven Sm proteins forms a complex that binds to several RNAs in metazoans. All Sm proteins contain a sequence signature, the Sm domain, also found in two yeast Sm-like proteins associated with the U6 snRNA. We have performed database searches revealing the presence of 16 proteins carrying an Sm domain in the yeast genome. Analysis of this protein family confirmed that seven of its members, encoded by essential genes, are homologues of metazoan Sm proteins. Immunoprecipitation revealed that an evolutionarily related subgroup of seven Sm-like proteins is directly associated with the nuclear U6 and pre-RNase P RNAs. The corresponding genes are essential or required for normal vegetative growth. These proteins appear functionally important to stabilize U6 snRNA. The two last yeast Sm-like proteins were not found associated with RNA, and neither was essential for vegetative growth. To investigate whether U6-associated Sm-like protein function is widespread, we cloned several cDNAs encoding homologous human proteins. Two representative human proteins were shown to associate with U6 snRNA containing complexes. We also identified archaeal proteins related to Sm and Sm like proteins. Our results demonstrate that Sm and Sm-like proteins assemble in at least two functionally conserved complexes of deep evolutionary origin. PMID- 10369685 TI - Partial purification of the yeast U2 snRNP reveals a novel yeast pre-mRNA splicing factor required for pre-spliceosome assembly. AB - We have partially purified the U2 snRNP of Saccharomyces cerevisiae. Identification of some proteins consistently found in the purified fractions by nanoelectrospray mass spectrometry indicated the presence of a novel splicing factor named Rse1p. The RSE1 gene is essential and codes for a 148.2 kDa protein. We demonstrated that Rse1p associates specifically with U2 snRNA at low salt concentrations. In addition, we showed that Rse1p is a component of the pre spliceosome. Depletion of Rse1p and analysis of a conditional mutant indicated that Rse1p was required for efficient splicing in vivo. In vitro Rse1p is required for the formation of pre-spliceosomes. Database searches revealed that Rse1p is conserved in humans and that it belongs to a large protein family that includes polyadenylation factors and DNA repair proteins. The characteristics of Rse1p suggest that its human homologue could be a subunit of the SF3 splicing factor. PMID- 10369686 TI - Structure-specific tRNA-binding protein from the extreme thermophile Aquifex aeolicus. AB - The genome of the bacterium Aquifex aeolicus encodes a polypeptide which is related to a small portion of a sequence found in one prokaryotic and two eukaryotic tRNA synthetases. It also is related to a portion of Arc1p, a tRNA binding protein believed to be important for nuclear trafficking of tRNAs. Here we cloned, expressed and purified the 111 amino acid polypeptide (designated Trbp111) and showed by ultracentrifugation analysis that it is a stable dimer in solution. The protein was also crystallized in a monoclinic lattice. X-ray diffraction analysis at 2.8 A resolution revealed a prominent non crystallographic 2-fold axis, consistent with the presence of a symmetric homodimeric structure. Band-shift analysis with polyacrylamide gels showed that the dimer binds tRNAs, but not RNA duplexes, RNA hairpins, single-stranded RNA nor 5S rRNA. Complex formation with respect to tRNA is non-specific, with a single tRNA bound per dimer. Thus, Trbp111 is a structure-specific tRNA-binding protein. These results and other considerations raise the possibility that Trbp111 is a tRNA-specific chaperone which stabilizes the native L-shaped fold in the extreme thermophile and which has been incorporated into much larger tRNA binding proteins of higher organisms. PMID- 10369687 TI - Deficiency in Msh2 affects the efficiency and local sequence specificity of immunoglobulin class-switch recombination: parallels with somatic hypermutation. AB - During maturation of the immune response, IgM+ B cells switch to expression of one of the downstream isotypes (IgG, A or E). This class switching occurs by region-specific recombination within the IgH locus through an unknown mechanism. A lack of switch recombination in mice deficient in components of the DNA dependent protein kinase (DNA-PK)-Ku complex has pointed to a role for non homologous end joining. Here we characterize a switching defect in mice lacking a protein involved in DNA mismatch recognition. Mice deficient in Msh2 give diminished IgG (but not IgM) responses following challenge with both T cell dependent and T cell-independent antigens. This appears to reflect a B cell intrinsic defect since B cells from Msh2-deficient mice also exhibit impaired switching (but not blasting or proliferation) on in vitro culture with lipopolysaccharide. Furthermore, those switches that do occur in Msh2-deficient B cells reveal a shift in the distribution of recombination sites used: the breakpoints are more likely to occur in consensus motifs. These results, which intriguingly parallel the effects of Msh2 deficiency on hypermutation, suggest a role for Msh2 in the mechanics of class-switch recombination. PMID- 10369688 TI - Xeroderma pigmentosum variant (XP-V) correcting protein from HeLa cells has a thymine dimer bypass DNA polymerase activity. AB - Xeroderma pigmentosum variant (XP-V) represents one of the most common forms of this cancer-prone DNA repair syndrome. Unlike classical XP cells, XP-V cells are normal in nucleotide excision repair but defective in post-replication repair. The precise molecular defect in XP-V is currently unknown, but it appears to be a protein involved in translesion synthesis. Here we established a sensitive assay system using an SV40 origin-based plasmid to detect XP-V complementation activity. Using this system, we isolated a protein from HeLa cells capable of complementing the defects in XP-V cell extracts. The protein displays novel DNA polymerase activity which replicates cyclobutane pyrimidine dimer-containing DNA templates. The XPV polymerase activity was dependent on MgCl2, sensitive to NEM, moderately sensitive to KCl, resistant to both aphidicolin and ddTTP, and not stimulated by PCNA. In glycerol density gradients, the activity co-sedimented with a 54 kDa polypeptide at 3.5S, indicating that the monomeric form of this polypeptide was responsible for the activity. The protein factor corrected the translesion defects of extracts from three XPV cell strains. Bypass DNA synthesis by the XP-V polymerase occurred only in the presence of dATP, indicating that it can incorporate only dATP to bypass a di-thymine lesion. PMID- 10369689 TI - M.(phi)BssHII, a novel cytosine-C5-DNA-methyltransferase with target-recognizing domains at separated locations of the enzyme. AB - In all cytosine-C5-DNA-methyltransferases (MTases) from prokaryotes and eukaryotes, remarkably conserved amino acid sequence elements responsible for general enzymatic functions are arranged in the same canonical order. In addition, one variable region, which includes the target-recognizing domain(s) (TRDs) characteristic for each enzyme, has been localized in one region between the same blocks of these conserved elements. This conservation in the order of conserved and variable sequences suggests stringent structural constraints in the primary structure to obtain the correct folding of the enzymes. Here we report the characterization of a new type of a multispecific MTase, M.(phiphi)BssHII, which is expressed as two isoforms. Isoform I is an entirely novel type of MTase which has, in addition to the TRDs at the conventional location, one TRD located at a non-canonical position at its N-terminus. Isoform II is represented by the same MTase, but without the N-terminal TRD. The N-terminal TRD provides HaeII methylation specificity to isoform I. The TRD is fully functional when engineered into either the conventional variable region of M.(phiphi)BssHII or the related monospecific M.phi3TII MTase. The implications of this structural plasticity with respect to the evolution of MTases are discussed. PMID- 10369691 TI - Microsatellite instability. PMID- 10369692 TI - A key cytokine unlocks the door. PMID- 10369690 TI - Progressive cis-inhibition of telomerase upon telomere elongation. AB - In yeast, the constant length of telomeric DNA results from a negative regulation of telomerase by the telomere itself. Here we follow the return to equilibrium of an abnormally shortened telomere. We observe that telomere elongation is restricted to a few base pairs per generation and that its rate decreases progressively with increasing telomere length. In contrast, in the absence of telomerase or in the presence of an over-elongated telomere, the degradation rate linked to the succession of generations appears to be constant, i.e. independent of telomere length. Together, these results indicate that telomerase is gradually inhibited at its site of action by the elongating telomere. The implications of this finding for the dynamics of telomere length regulation are discussed in this study. PMID- 10369693 TI - Glutamine in parenteral nutrition: more food for thought. PMID- 10369694 TI - Sex and drugs and HCV? PMID- 10369695 TI - Cryoglobulinaemia in HCV infection: coming in from the cold. PMID- 10369696 TI - ERCP training: for the few, not for all. PMID- 10369697 TI - The gut as target organ for oral immunovaccination with allergen DNA: new hope for patients with anaphylactic reactions to food? PMID- 10369698 TI - What makes the dyspeptic patient feel ill? A cross sectional survey of functional health status, Helicobacter pylori infection, and psychological distress in dyspeptic patients in general practice. AB - BACKGROUND: Dyspepsia is prevalent in about 30% of the general population in Europe, but only 25% of people with complaints consult their general practitioner. AIMS: To study the relation between the severity of dyspeptic complaints and the health status of patients presenting to the general practitioner; and the relation with patient characteristics, Helicobacter pylori infection, and psychological distress. METHODS: A cross sectional, general practice based survey of 360 unselected primary care dyspeptic patients from 92 general practices in The Netherlands was conducted. Symptom severity was measured using a validated symptom score, H pylori using a whole blood test, and psychological distress using the GHQ-12 test. Functional health status was assessed using the COOP/Wonca charts. RESULTS: Symptoms lasting more than three months and presence of relevant psychological distress were both associated with higher levels of dyspepsia. H pylori infection, frequency of symptoms, and age had no influence on dyspepsia severity. Severity of dyspepsia and psychological distress, but not H pylori infection or duration of symptoms, affected health status univariately. Dyspepsia correlated with general health, daily activities, and social activities. In logistic modelling, health status was far better predicted by psychological distress than by dyspepsia severity. CONCLUSION: The relation between dyspeptic symptom severity and health status is limited. H pylori infection relates neither to functional health status, nor to intensity of dyspepsia. Psychological distress is a major determinant of impaired health of dyspeptic patients in general practice and may be the clue to improvement of health status in many dyspeptic patients. PMID- 10369699 TI - Antibodies to human gastric epithelial cells and heat shock protein 60 in Helicobacter pylori positive mucosa associated lymphoid tissue lymphoma. AB - BACKGROUND: Development of gastric mucosa associated lymphoid tissue (MALT) lymphoma is thought to be closely associated with host immune reactions to Helicobacter pylori. AIM: To investigate humoral immune responses in patients with MALT lymphoma to antigens shared by H pylori and human gastric epithelial cells. METHODS: Sera were obtained from H pylori positive patients with MALT lymphoma (n = 11) or other gastroduodenal diseases (peptic ulcer, n = 40; non ulcer dyspepsia, n = 20) and from H pylori negative healthy control subjects (n = 10). Antibodies to HGC-27 human gastric epithelial cells and human recombinant heat shock protein (Hsp) 60 were examined using an enzyme linked immunosorbent assay (ELISA) and immunoblotting. RESULTS: Antibody titres to HGC-27 cells were significantly elevated in H pylori positive patients with MALT lymphoma when compared with titres in patients with other gastroduodenal diseases and in healthy subjects. Immunoblotting of sera from patients with MALT lymphoma often detected a band with a molecular mass corresponding to Hsp60, and both ELISA and immunoblotting showed elevated antibody titres to the recombinant human Hsp60. Antigenic similarity between Hsp60 and H pylori HspB was documented by immunoblotting experiments. CONCLUSIONS: Autoantibodies reactive with host gastric epithelial cells are often increased in MALT lymphoma, and Hsp60 is a major target antigen. Immune responses induced by immunological cross reactivity between H pylori HspB and human Hsp60 in gastric epithelium may be involved in the development of MALT lymphoma. PMID- 10369700 TI - Regulation of tumour necrosis factor (TNF) induced apoptosis by soluble TNF receptors in Helicobacter pylori infection. AB - BACKGROUND: Tumour necrosis factor (TNF) is a predominant cytokine produced in the gastric mucosa of patients with Helicobacter pylori infection. TNF induces apoptosis in a variety of cells. The soluble TNF receptors (sTNF-Rs) can be divided into sTNF-RI and sTNF-RII, both of which inhibit TNF activity. However, their precise mechanisms remain unclear. AIM: To investigate the role of sTNF-Rs in H pylori infection. METHODS: In 40 patients, production of TNF and sTNF-Rs in gastric mucosa was measured using biopsy specimens. In addition, in gastric epithelial cells, sTNF-R release in response to TNF and the protective effect of sTNF-Rs against the cytotoxic and apoptotic activities of TNF were examined. RESULTS: TNF and sTNF-R expression was significantly higher in H pylori positive than H pylori negative patients. TNF dose-dependently induced sTNF-RI release from gastric epithelial cells. sTNF-RII was also released from the cells. TNF decreased cell viability, but the effect was very small. A combination of anti sTNF-RI and anti-sTNF-RII monoclonal antibodies significantly increased TNF induced cytotoxicity and apoptosis of gastric epithelial cells. CONCLUSIONS: These results show that sTNF-Rs are actively produced in H pylori infected gastric mucosa. sTNF-Rs appear to protect gastric epithelial cells from TNF induced apoptosis in H pylori infection. PMID- 10369701 TI - Hereditary colorectal cancer in the general population: from cancer registration to molecular diagnosis. AB - BACKGROUND: Hereditary non-polyposis colorectal cancer (HNPCC) is one of the most common inherited disorders predisposing to cancer. The genes responsible for the disease have recently been cloned and characterised; their mutations induce a generalised genomic instability which is particularly evident at microsatellite loci (replication error (RER)+ phenotype). AIMS: To investigate how to select individuals and families in the general population who should be screened for constitutional mutations predisposing to colorectal cancer. PATIENTS/METHODS: Between 1984 and 1995, 1899 colorectal malignancies in 1831 patients were registered, and in 1721 of these (94%), family trees could be obtained. Patients and families were classified into five categories according to a more or less likely genetic basis: HNPCC; "suspected" HNPCC; juvenile cases; aspecific cancer aggregation; sporadic cases. In 18 families with HNPCC as well as in 18 with suspected HNPCC, microsatellite instability in tumour tissues and constitutional mutations of two DNA mismatch repair genes (MSH2 and MLH1) could be evaluated. RER status was studied with five markers (BAT40, D2S123, D18S57, D17S787, and BAT26) in paraffin embedded tissues. Germline mutations of MSH2 or MLH1 genes were assessed on DNA and RNA extracted from lymphomonocytic cells, using reverse transcription polymerase chain reaction, single strand conformation polymorphism analysis, and direct DNA sequencing. RESULTS: HNPCC represented 2.6% and suspected HNPCC 4.6% of all registered colorectal neoplasms. Eleven out of 18 HNPCC families (61%) showed microsatellite instability as opposed to four (of 18) suspected HNPCC (22%; p<0.02). Three germline mutations (two in MSH2 and one in MLH1 gene) were found in three different large HNPCC families, whereas no mutations were detected in suspected HNPCC. CONCLUSIONS: In this study of cancer genetic epidemiology, data from a tumour registry were analysed and this ultimately led to the identification and selection of families that should be tested for mutator gene mutations. With the use of a population based approach, the incidence of mutations was appreciably lower than previously reported and limited to families with full blown HNPCC. It is possible that in most families with a clinical spectrum of HNPCC (or suspected HNPCC) other DNA mismatch repair genes are involved in the pathogenesis of the disease. PMID- 10369702 TI - Survival after rectal cancer: differences between hospital catchment areas. A nationwide study in Sweden. AB - BACKGROUND: The quality of rectal cancer surgery at small units has been debated. No national studies of this issue have been undertaken and most studies have been based on insufficient data to clarify the controversy. It has been claimed that observed differences in outcomes between specialised centres and smaller hospitals are confounded by differences in stage/severity. AIM: To compare survival after rectal cancer between hospital catchment areas. PATIENTS: All patients with rectal cancer notified to the Swedish Cancer Register in 1973-1992 (n = 30 811) were followed up by record linkage to the nationwide Death Register. METHODS: Relative survival-that is, ratio of observed to expected survival-was computed as a measure of excess mortality attributable to rectal cancer. Multivariate analysis was then performed to estimate the independent effects of hospital catchment area categories and age, year of diagnosis, and duration of follow up. RESULTS: One year relative survival among rectal cancer patients residing in catchment areas of large regional hospitals was 76%, compared with 72% for small local hospitals (p<0.001). A difference was already noted after 30 days and remained five years after diagnosis. Relative survival improved considerably overall, but the differences between catchment area categories persisted. These were not reduced by adjustment for age, time after diagnosis, or time period in multivariate models. CONCLUSION: The differences in outcome between catchment area categories could not be explained by differences in age, time period, or duration of follow up after diagnosis. They are unlikely to be explained by differences between catchment area populations with regard to the average stage of the disease at which symptoms lead to diagnosis. The differences may therefore be attributable to different strategies for diagnosing and managing patients with rectal cancer. PMID- 10369703 TI - Human right and left colon differ in epithelial cell apoptosis and in expression of Bak, a pro-apoptotic Bcl-2 homologue. AB - BACKGROUND: Propensity to colonic neoplasia differs between the right and left colon. AIMS: To examine whether this difference may be related to regional differences in epithelial apoptosis, in expression of a proapoptotic regulatory protein, Bak, and in proliferation. PATIENTS: Individuals with no history of colorectal neoplasia. METHODS: Archival blocks of colorectal tissues were immunostained for proliferating cells (antibody to Ki-67 antigen), and Bak expression (polyclonal antiserum). Cells containing DNA strand breaks, a marker of apoptosis, were identified by terminal deoxyuridine nucleotidyl nick end labelling (TUNEL). RESULTS: There were fewer TUNEL positive epithelial cells in the right colon (mean 1.2 (SE 0.1)% of all epithelial cells) than the left colon (2.2 (0.1)%, p<0.0001) or rectum (2.2 (0.3)%, p<0.05). Bak expression was less common in the right colon (mean 46 (2.3)% of epithelial cells immunoreactive) than the left colon (66 (2.7)%, p<0.0001), or rectum (67 (2.3)%, p<0.001). Bak expression and TUNEL positivity were highly positively correlated (p<0.0001). In contrast to apoptosis, mean whole crypt proliferation labelling index was similar throughout the colorectum (right colon: 15.6 (3.2)%; left colon: 13. 5 (1.2)%; rectum: 13.3 (2.3)%). CONCLUSION: The percentage of proliferating colonic epithelial cells is constant throughout the colon, but fewer epithelial cells undergo Bak mediated apoptosis in the right than in the left colon or rectum. This suggests that colonocytes may be lost by methods other than apoptosis in the right colon. PMID- 10369704 TI - Induction of retinoic acid receptor beta mediates growth inhibition in retinoid resistant human colon carcinoma cells. AB - BACKGROUND: The molecular mechanisms underlying the differential sensitivity of human colon carcinoma cells to retinoid mediated growth inhibition are poorly understood. AIM: To identify the intracellular mechanisms responsible for resistance against retinoid mediated growth inhibition in human colon carcinoma cells. METHODS: Anchorage independent growth of the human colon carcinoma cell lines HT29 and LoVo was determined by a human tumour clonogenic assay. Retinoid receptor expression was evaluated by reverse transcription polymerase chain reaction and northern blotting. Retinoid mediated transactivation was assessed by transient transfection of a pTK::betaREx2-luc reporter construct. Retinoid receptor overexpression was achieved by selecting stably transfected cell clones. RESULTS: Retinoid treatment resulted in profound dose dependent growth inhibition in HT29 cells, while LoVo cells were unaffected. The two cell lines express identical patterns of nuclear retinoid receptor mRNA transcripts. However, on retinoid treatment, retinoic acid receptor beta gene expression was upregulated only in retinoid sensitive HT29 cells, but not in retinoid resistant LoVo cells. In accordance, stable overexpression of retinoic acid receptor beta but not alpha or gamma conferred retinoid mediated growth inhibition on LoVo cells. CONCLUSION: Induction of retinoic acid receptor beta expression is required and sufficent to confer retinoid mediated growth inhibition on human colon carcinoma cells. PMID- 10369705 TI - Laterality effects of human pudendal nerve stimulation on corticoanal pathways: evidence for functional asymmetry. AB - BACKGROUND: Although motor and sensory pathways to the human external anal sphincter are bilateral, a unilateral pudendal neuropathy may still disrupt anal continence. Anal continence can, however, be preserved despite unilateral pudendal damage, and so to explain those differing observations, we postulated that pudendal innervation might be asymmetric. AIMS: To explore the individual effects of right and left pudendal nerve stimulation on the corticofugal pathways to the human external anal sphincter and thus assess evidence for functional asymmetric pelvic innervation. METHODS: In eight healthy subjects, anal sphincter electromyographic responses, evoked to transcranial magnetic stimulation of the motor cortex, were recorded 5-500 msec after digital transrectal electrical conditioning stimuli applied to each pudendal nerve. RESULTS: Right or left pudendal nerve stimulation evoked anal responses of similar latencies but asymmetric amplitudes in six subjects: dominant responses (>50% contralateral side) from the right pudendal in four subjects and from the left in two. Cortical stimulation also evoked anal responses with amplitude 448 (121) microV and latency 20.9 (1.1) msec. When cortical stimulation was preceded by pudendal nerve stimulation, the cortical responses were facilitated at interstimulus intervals of 5-20 msec. Dominant pudendal nerve stimulation induced greater facilitation of the cortically evoked responses than the non-dominant nerve. CONCLUSIONS: Cortical pathways to the external anal sphincter are facilitated by pudendal nerve conditioning, in an asymmetric manner. This functional asymmetry may explain the presence and absence of anal incontinence after unilateral pudendal nerve injury. PMID- 10369706 TI - Laser Doppler measurement of rectal mucosal blood flow. AB - BACKGROUND: Gut mucosal blood flow measurement is used to study a variety of disorders and possibly extrinsic neural function. AIMS: To determine optimal measurement criteria and validate this technique as a measure of level of activity of extrinsic autonomic gut innervation. METHODS: In 26 healthy volunteers a laser Doppler mucosal probe was applied 10 cm from the anus. Response to inhaled salbutamol 200 microgram and ipratropium 40 microgram, intravenous metoprolol 2.5 mg, and direct sacral nerve electrostimulation (in nine incontinent patients) was also studied. RESULTS: The coefficient of variation for subjects studied under identical conditions on two, three, and four days was 0.06, 0.05, and 0.06, respectively. Mean mucosal blood flow increased after a standard meal. Blood flow decreased for 15 minutes after smoking and returned to baseline at 30 minutes. Fasted measurements at 0900, 1200, 1600, and 2200 were similar. There was a negative correlation between blood flow and body size but not age. Follicular phase mucosal flow was less and more reproducible than luteal. Mucosal blood flow was highest in men and lowest in postmenopausal women. Inhaled salbutamol did not change blood flow; ipratropium significantly reduced, and metoprolol and sacral nerve stimulation increased flow. CONCLUSIONS: Measurement of gut mucosal blood flow by laser Doppler flowmetry is highly reproducible. Eating, smoking, body size, sex, ovulatory status, and menstrual phase influence blood flow. Changes in mucosal blood flow induced by autonomically active drugs and nerve stimulation confirm the role of the mucosal microcirculation as a measure of extrinsic nerve activity. PMID- 10369707 TI - Small intestinal transit, absorption, and permeability in patients with AIDS with and without diarrhoea. AB - BACKGROUND: Diarrhoea in AIDS is associated with anorexia and weight loss. The importance of gastrointestinal transit in such symptoms has not been addressed. AIMS: To assess jejunal to caecal transit times in subjects with AIDS related diarrhoea and weight loss and correlate these with measures of absorptive capacity and intestinal permeability. METHODS: Jejunal to caecal transit times were assessed in 20 seronegative controls and 60 HIV seropositive subjects from serum analysis of 3-O-methyl-D-glucose and sulphapyridine after ingestion of the monosaccharide and sulphasalazine in aqueous solution. The method also allows an estimation of gastric emptying times for liquids. Intestinal absorptive capacity and permeability were assessed by a combined test using 3-O-methyl-D-glucose, D xylose, L-rhamnose, and lactulose. RESULTS: Gastric emptying was significantly delayed in all groups of patients with AIDS. Mean jejunal to caecal transit times were not significantly different between controls (246 (62) minutes) and patients without diarrhoea (AIDS, well: 278 (103) minutes; AIDS, wasting: 236 (68) minutes), cytomegalovirus colitis (289 (83) minutes), pathogen negative diarrhoea (192 (100) minutes), or microsporidiosis (190 (113) minutes), although 30% of patients had values below the control range. Patients with cryptosporidiosis differed significantly from controls (135 (35) minutes, p<0.0001), seven of 10 having rapid transit times. Absorptive capacity was reduced and intestinal permeability significantly increased in AIDS, but did not correlate significantly with transit times. CONCLUSION: Small bowel transit is accelerated in many patients with AIDS, particularily in protozoal diarrhoea, but is not the sole explanation for malabsorption of monosaccharides. PMID- 10369708 TI - Gastric pathology in patients with common variable immunodeficiency. AB - BACKGROUND/AIMS: Common variable immunodeficiency (CVID) is an immunological disorder characterised by defective antibody production. Patients with CVID have a high risk of gastric cancer. It has been suggested that gastric cancer results from an interaction between environmental factors and a genetic predisposition. The role of Helicobacter pylori as an environmental factor in gastric carcinogenesis is of current interest. Moreover, p53 gene mutations have been reported in gastric cancer. This study focuses on the gastric pathology of patients with CVID and correlation with H pylori infection. METHODS: Thirty four consecutive dyspeptic patients with CVID (mean age 49.6 years, range 14-72; 17 men) were included in the study. An upper gastrointestinal endoscopy was performed and biopsy specimens were taken from the antrum, incisura angularis, and gastric body. Biopsies were used for histological assessment, to identify the presence of H pylori, and to evaluate p53 overexpression. RESULTS: H pylori infection was detected in 14/34 (41%) patients. Chronic active gastritis involving both antrum and body was observed more frequently in H pylori positive (79%) than H pylori negative (20%) patients (p = 0.001). Similarly, a histological feature of multifocal atrophic gastritis was found more frequently in infected (50%) than uninfected patients (10%) (p = 0.012). In addition, one case of gastric adenocarcinoma and another of notable dysplasia were observed in the H pylori positive group. Overexpression of p53 was found in six (18%) patients, including one with normal gastric mucosa. CONCLUSIONS: It can be hypothesised that both H pylori and p53 alterations play a role in the gastric carcinogenesis of patients with CVID. PMID- 10369709 TI - A double blind, randomised, controlled trial of glutamine supplementation in parenteral nutrition. AB - BACKGROUND AND AIMS: To determine whether the inclusion of 20 g free glutamine as part of the nitrogen source of parenteral feeds reduces length of hospital stay or mortality. METHODS: In a randomised, double blind, controlled trial in 168 patients clinically accepted for parenteral nutrition, standard feeds were compared with feeds in which 3.8 g of the total nitrogen was replaced with the equivalent 20 g glutamine. A minimum of 11 g nitrogen/day was used in all patients. Daily intakes of energy and nitrogen were determined using a validated computer protocol and were similar for the two groups. All feeds included trace elements, vitamins, electrolytes, and minerals. RESULTS: A total of 85 patients received a median of eight (interquartile range 5-13) daily feeds containing glutamine while 83 received a median of eight (5-15) standard feeds. No difference between groups was detected for infective complications. Twenty control patients and 14 who had received glutamine died during their hospital stay (NS). Median length of stay was 32 (23-52) days on glutamine, which was not significantly different from the control value of 35 (25-55) days. Glutamine was associated with a significant (p<0.03) reduction in length of stay in surgical patients (45 days (range 29-81) versus 30 days (range 19-54)). CONCLUSION: The benefit from glutamine supplementation of parenteral feeds as used in this trial has not been proved. Supplementation may have advantages in surgical patients and in haematological malignancy. Further trials are required. PMID- 10369710 TI - Saccharomyces boulardii upgrades cellular adaptation after proximal enterectomy in rats. AB - BACKGROUND: Saccharomyces boulardii is a non-pathogenic yeast which exerts trophic effects on human and rat small intestinal mucosa. AIMS: To examine the effects of S boulardii on ileal adaptation after proximal enterectomy in rats. METHODS: Wistar rats, aged eight weeks, underwent 60% proximal resection or transection and received by orogastric intubation either 1 mg/g body wt per day lyophilised S boulardii or the vehicle for seven days. The effects on ileal mucosal adaptation were assessed eight days after surgery. RESULTS: Compared with transection, resection resulted in mucosal hyperplasia with significant decreases in the specific and total activities of sucrase, lactase, and maltase. Treatment of resected animals with S boulardii had no effect on mucosal hyperplasia but did upgrade disaccharidase activities to the levels of the transected group. Enzyme stimulation by S boulardii was associated with significant increases in diamine oxidase activity and mucosal polyamine concentrations. Likewise, sodium dependent D-glucose uptake by brush border membrane vesicles, measured as a function of time and glucose concentration in the incubation medium, was significantly (p<0.05) increased by 81% and three times respectively in the resected group treated with S boulardii. In agreement with this, expression of the sodium/glucose cotransporter-1 in brush border membranes of resected rats treated with S boulardii was enhanced twofold compared with resected controls. CONCLUSION: Oral administration of S boulardii soon after proximal enterectomy improves functional adaptation of the remnant ileum. PMID- 10369711 TI - A randomised, double blind, multicentre trial of octreotide in moderate to severe acute pancreatitis. AB - BACKGROUND: The pharmacological inhibition of exocrine pancreatic secretion with the somatostatin analogue octreotide has been advocated as a specific treatment of acute pancreatitis. AIM: To investigate the efficacy of octreotide in acute pancreatitis in a randomised, placebo controlled trial. METHODS: 302 patients from 32 hospitals, fulfilling the criteria for moderate to severe acute pancreatitis within 96 hours of the onset of symptoms, were randomly assigned to one of three treatment groups: group P (n=103) received placebo, while groups O1 (n=98) and O2 (n=101) received 100 and 200 microg of octreotide, respectively, by subcutaneous injection three times daily for seven days. The primary outcome variable was a score composed of mortality and 15 typical complications of acute pancreatitis. RESULTS: The three groups were well matched with respect to pretreatment characteristics. An intent to treat analysis of all 302 patients revealed no significant differences among treatment groups with respect to mortality (P: 16%; O1: 15%; O2: 12%), the rate of newly developed complications, the duration of pain, surgical interventions, or the length of the hospital stay. A valid for efficacy analysis (251 patients) also revealed no significant differences. CONCLUSIONS: This trial shows no benefit of octreotide in the treatment of acute pancreatitis. PMID- 10369713 TI - Epidemiological and virological analysis of couples infected with hepatitis C virus. AB - BACKGROUND: If transmission of hepatitis C virus (HCV) infection through parenteral exposure is well documented, sexual transmission of HCV is still debated. AIMS: To perform extensive epidemiological and virological analysis in 24 couples in which each spouse was anti-HCV positive in order to delineate more precisely potential sexual transmission of HCV. PATIENTS: Twenty four couples in which each partner was anti-HCV positive. These 48 spouses were recruited in a liver unit by regular screening of spouses of index patients. METHODS: All 48 spouses completed an epidemiological questionnaire on risk factors for HCV. Qualitative detection of serum HCV RNA and determination of HCV type by genotyping and serotyping were performed. Sequence analysis of HCV strains by phylogenetic analysis was carried out in seven couples with concordant genotypes. RESULTS: The mean (SD) partnership duration was 12 (10) years. Serum HCV RNA was detected in both partners in 18 of the couples (75%) and in only one partner in six of the couples (25%). HCV typing showed concordant genotypes in 12 couples (50%), discordant genotypes in seven (29%), and in the other five couples (21%) only one spouse could be genotyped. Of the 48 spouses, 33 had a major risk factor for HCV transmission such as transfusion (n = 6) and intravenous drug use (n = 27). Eleven of the 12 couples infected with the same HCV genotype had at least one parenteral risk factor for viral transmission in both spouses. Whatever the genotype concordance, in most couples (75%), both spouses showed parenteral risk factors for viral transmission. Sequence analysis of HCV strains was possible in seven of 12 couples with identical genotype and showed different and identical isolates in four and three couples respectively. CONCLUSION: The study emphasises the risk of overestimating the importance of a very low sexual HCV transmission risk as against other, mainly parenteral, risk factors. PMID- 10369712 TI - Induction of TFF1 gene expression in pancreas overexpressing transforming growth factor alpha. AB - BACKGROUND/AIMS: Chronic pancreatitis is an inflammatory disease of the exocrine pancreas associated with extensive fibrosis, enlarged pancreatic ducts, acinar cell degeneration, and the formation of tubular complexes. The molecular and biochemical alterations associated with these histological changes are not kown. Generally, the new family of TFF peptides (formerly known as P-domain peptides or trefoil factors) is aberrantly expressed during chronic inflammatory diseases of the gastrointestinal tract. METHODS: Using human pancreatic tissues obtained from patients with chronic pancreatitis and murine pancreatic tissues obtained from transgenic mice overexpressing transforming growth factor alpha (TGF-alpha), the expression and cellular distribution of TFF1 was analysed using northern blot analysis, polymerase chain reaction (PCR), and immunohistochemistry. RESULTS: In the normal human pancreas, TFF1 was scarce, with only a few ducts exhibiting cytoplasmic TFF1 immunoreactivity. In contrast, human chronic pancreatitis tissue specimens exhibited strong TFF1 immunoreactivity in ductal cells, areas of ductal hyperplasia, and tubular complexes. Semiquantitative PCR analysis of TFF1 mRNA levels showed enhanced expression of TFF1 in the pancreas of patients with chronic pancreatitis. Furthermore, TFF1 mRNA levels were detectable in the pancreas in four of five transgenic mice overexpressing TGF-alpha. In contrast, four of five wild type mice did not exhibit a TFF1 mRNA transcript. In addition, while no specific TFF1 immunoreactivity was present in the pancreas of the wild type mice, ductal epithelial cells and duct-like tubular complexes in the pancreas of the transgenic mice overexpressing TGF-alpha exhibited pronounced TFF1 immunoreactivity. CONCLUSIONS: Ductal cells and tubular complexes in pancreatic fibrosis express TFF1. As the 5'-flanking region of TFF1 contains an epidermal growth factor responsive enhancer region and the expression of epidermal growth factor and TGF-alpha is enhanced in pancreatic fibrosis, the enhanced expression of TFF1 in pancreatic fibrosis may be mediated by TGF-alpha. PMID- 10369714 TI - Relation between severity of liver disease and renal oxygen consumption in patients with cirrhosis. AB - BACKGROUND: Worsening cirrhosis may lead to increased renal O2 metabolism caused by activation of neurohumoral antinatriuretic substances. AIMS: To evaluate the relation between the severity of liver disease, sodium excretion, and neurohumoral antinatriuretic substances on the one hand and renal O2 metabolism on the other in patients with cirrhosis. METHODS: Renal O2 consumption and haemodynamics as well as plasma concentrations of noradrenaline, renin, and aldosterone were measured. Investigations were performed in 14 patients with Pugh's grade A, 43 with grade B, and 29 with grade C liver disease. RESULTS: Renal O2 consumption significantly increased with the severity of cirrhosis (grade A, 8.9 (1.6); grade B, 15.5 (1.3); grade C, 18.0 (1.5) ml/min/m2). Plasma concentrations of noradrenaline, renin, and aldosterone significantly increased while mean arterial presssure and systemic vascular resistance significantly decreased with the severity of the disease. A significant inverse correlation was found between renal O2 consumption and sodium excretion. A significant direct correlation was found between plasma levels of noradrenaline and aldosterone on the one hand and renal O2 consumption on the other. Renal blood flow and the glomerular filtration rate did not differ significantly between patients with grade C and grade A or B disease. CONCLUSIONS: This study shows for the first time that, in patients with cirrhosis, worsening of the disease is associated with an increase in renal O2 consumption. The results suggest that increased renal O2 consumption is due to renal tubular sodium retention caused by increased levels of neurohumoral antinatriuretic substances. This neurohumoral activation is related to cirrhosis induced vasodilation. PMID- 10369715 TI - Response to interferon alpha treatment and disappearance of cryoglobulinaemia in patients infected by hepatitis C virus. AB - BACKGROUND: Mixed cryoglobulinaemia is closely associated with hepatitis C virus (HCV) infection. AIM: To assess in a prospective open study the efficiency of interferon alpha treatment of cryoglobulinaemia, as reflected by the disappearance of cryoglobulins and clinical manifestations of the disease, and to analyse the factors predictive of a response to interferon. METHOD: Eighty seven consecutive patients with chronic hepatitis C treated for the first time with interferon at a dose of 3 x 10(6) international units three times a week for six months were studied. Forty three patients had cryoglobulins, which were responsible for clinical manifestations in 12. RESULTS: At the end of interferon treatment, cryoglobulins had disappeared in 39% of the patients. A clinical improvement (except for neuropathies) was observed in all patients. Six months after interferon treatment was stopped, the same rate of response (normal alanine aminotransferase values and undectable HCV RNA) was observed in patients with or without cryoglobulins. Only 14% of patients still had undetectable cryoglobulins, and all of them also had undetectable serum HCV RNA. The disappearance of cryoglobulins was found less frequently in patients with clinical symptoms than in asymptomatic ones, but the difference was not significant. Sustained responders were more often men, infected by genotype 2 or 3, with a lower pretreatment viral load. CONCLUSION: The presence of cryoglobulins does not seem to affect the response to interferon in HCV infected patients. The improvement in cryoglobulinaemia is strongly associated with a virological response, reinforcing the hypothesis of a direct role for HCV in the pathogenesis of this disease. PMID- 10369716 TI - A prospective study of the causes of notably raised aspartate aminotransferase of liver origin. AB - BACKGROUND AND AIMS: To ascertain the causes of raised aspartate aminotransferase (AST) presumed to be of hepatic origin in two hospitals and the local community served by a centralised biochemistry laboratory. METHODS: From June 1996 to February 1997 all patients with AST greater than 400 U/l were identified by the biochemistry laboratory; the patients' clinical records were studied to determine the diagnosis, the clinical outcome, and whether the raised AST and its significance had been noted. RESULTS: A total of 137 patients with a hepatic cause for the raised AST were found. The cause of the raised AST was hepatic ischaemia/hypoxia in 68, pancreatobiliary disease in 33, primary hepatocellular disease in 23, hepatic malignancy in five, and hepatic haematoma in one. In seven patients the diagnosis was unclear. The overall mortality was high (22%) with the highest mortality in the hepatic ischaemia group (37%). The recording and interpretation of the causes of raised AST was poor with only 48% having the correct diagnosis. In 38% the raised AST was apparently not noticed by the attending clinicians. CONCLUSIONS: The commonest cause of a hepatitis like biochemical picture was hepatic hypoxia (50%) followed by pancreatobiliary disease (24%). Drug induced hepatic necrosis (8.8%) was uncommon and viral hepatitis was rare (3.6%). AST concentrations returned towards normal most rapidly in patients with hepatic hypoxia and calculous biliary obstruction. Hepatitis, viral or otherwise, is an uncommon cause of a typical hepatitic biochemical result in this community. PMID- 10369718 TI - Intestinal ganglioneuromatosis and multiple endocrine neoplasia type 2B: implications for treatment. AB - Three infants, who presented with intestinal obstruction due to diffuse transmural intestinal ganglioneuromatosis, are described. Mutation analysis of exon 16 of the RET proto-oncogene revealed germline M918T and thus, a molecular diagnosis of multiple endocrine neoplasia type 2B (MEN 2B). Two infants developed medullary carcinoma of the thyroid. The third had a prophylactic thyroidectomy despite no obvious thyroid masses and normal calcitonin concentrations, but microscopic multifocal medullary carcinoma was found on histological examination. Early recognition of intestinal ganglioneuromatosis with germline RET M918T mutation in pseudo-Hirschsprung's disease is an indication for prophylactic thyroidectomy. PMID- 10369717 TI - All-trans and 9-cis retinoic acid alter rat hepatic stellate cell phenotype differentially. AB - BACKGROUND: Hepatic stellate cells exert specific functions in the liver: storage of large amounts of retinyl esters, synthesis and breakdown of hepatic extracellular matrix, secretion of a variety of cytokines, and control of the diameter of the sinusoids. AIMS: To examine the influence of all-trans retinoic acid (ATRA) and 9-cis retinoic acid (9RA) on extracellular matrix production and proliferation of activated hepatic stellate cells. METHODS: Cells were isolated using collagenase/pronase, purified by centrifugation in nycodenz, and cultured for two weeks. At this time point the cells exhibited the activated phenotype. Cells were exposed to various concentrations of ATRA and 9RA. The expression of procollagens I, III, and IV, of fibronectin and of laminin were analysed by immunoprecipitation and northern hybridisation. RESULTS: ATRA exerted a significant inhibitory effect on the synthesis of procollagens type I, III, and IV, fibronectin, and laminin, but did not influence stellate cell proliferation, whereas 9RA showed a clear but late effect on proliferation. 9RA increased procollagen I mRNA 1.9-fold, but did not affect the expression of other matrix proteins. CONCLUSION: Results showed that ATRA and 9RA exert different, often contrary effects on activated stellate cells. These observations may explain prior divergent results obtained following retinoid administration to cultured stellate cells or in animals subjected to fibrogenic stimuli. PMID- 10369719 TI - Small, depressed lesions of the large bowel: a normal finding at endoscopy. PMID- 10369721 TI - Structured training and assessment in ERCP has become essential for the Calman era. PMID- 10369724 TI - Interpretation of absorptiometric bone mass measurements in the growing skeleton: issues and limitations. PMID- 10369720 TI - Molecules controlling lymphocyte migration to the gut. PMID- 10369725 TI - Recommendations for the registration of drugs intended for use in the treatment of male osteoporosis. PMID- 10369726 TI - Confocal laser scanning microscopy: A nondestructive subsurface histotomography of healthy human bone. AB - Microscopy of bony tissue usually requires special treatment for decalcification and processing of thin sections. Confocal laser scanning microscopy (CLSM) allows the nondestructive histotomography of organic hard tissue. The aim of this study was to visualize healthy human bone structures and to correlate identical areas in CLSM and conventional light microscopy. Each sample of healthy human lower jaw (n = 20) was divided into three parts: (1) fresh, untreated bony blocks studied by CLSM; (2) MMA-embedded thin sections (without decalcification), HE stained and studied by CLSM and conventional light microscopy (correlation of identical areas); (3) decalcificated, HE stained, histological sections studied by conventional light microscopy. In untreated bony blocks, microstructures such as osteocytes and lamellae were identified by CLSM. These structures could be correlated with conventional light microscopy. In CLSM, subcellular structures cannot yet be interpreted, whereas cytoplastic processes of osteocytes were seen with high contrast. With CLSM, nondestructive histology of cortical bone can be obtained. The risk of artifacts due to pretreatment is minimized, and subsurface visualization does not affect the interpretation. PMID- 10369727 TI - Abnormal mineral composition of osteogenesis imperfecta bone as determined by electron probe X-ray microanalysis on conventional and cryosections. AB - Osteogenesis imperfecta (OI) is a genetic disorder of the connective tissue characterized by frequent bone fractures. The cause of bone fragility is still unknown even though substantial work on collagen has been done. We measured the calcium to phosphorus ratio (Ca/P) of bone mineral from 35 OI bone samples and 25 age- and site-matched control specimens, using electron probe X-ray microanalysis in the transmission electron microscope. Ultra-thin cryosections and conventionally prepared resin sections were used. Cryo-ultramicrotomy avoids any possible artifactual demineralization that may occur in conventional aqueous media. The Ca/P ratio obtained by these two methods was compared and there was no statistical difference between them. The results were differentiated according to the clinical types of OI for the first time. The Ca/P ratio of OI bone mineral was lower than normal in both resin and cryosections, and mirrored the severity of the disease. OI type II had the lowest ratio (Ca/P = 1.49) compared with normal age- and site-matched controls (Ca/P = 1.69). This abnormal mineral composition in OI type II could be a contributory factor to bone fragility in OI bone. PMID- 10369728 TI - Early postmenopausal bone loss is prevented by estrogen and partially by 1alpha OH-vitamin D3: therapeutic effects of estrogen and/or 1alpha-OH-vitamin D3. AB - A total of 79 Japanese women who were within 5 years of menopause were randomly assigned 1alpha-hydroxyvitamin D3 [1alpha(OH)D3] 1.0 microg/day, conjugated estrogens 0.625 mg/day, a combination of both, or control (no treatment). Lumbar spine and proximal femur bone mineral density (BMD) and biochemical indices were monitored over 2 years. In the 1alpha(OH)D3-treated group, there was a nonsignificant decrease in lumbar spine BMD compared with controls, and no significant loss in the femoral neck compared with controls. In the estrogen treated group, there was a nonsignificant increase in spine BMD (+2.17% in the first year and +1.71% in the second year), and no loss in femoral neck BMD. The combination of conjugated estrogens +1alpha(OH)D3 was more effective in increasing BMD in the spine (+3. 68% in the first year and +3.63% in the second year) and femur (+2. 56% in the first year and +4.44% in the second year) BMD. There was a significant difference in lumbar spine BMD in both the first and second years between the combination-treated group and the 1alpha(OH)D3-treated and control groups (P < 0.01). Serum osteocalcin (OC) significantly decreased in the combination-treated group (-23.8% in the first year) and the estrogen-treated group (-37. 6% and -41.2% at 6 and 18 months, respectively), and serum alkaline phosphatase (Alp) decreased significantly in the first year in the combination treated (-31.5%), estrogen-treated (-27.3%), and 1alpha(OH)D3-treated (-7.9%) groups, whereas serum OC increased (+45. 4% in the first year) in women without treatment. The results of this study indicate that early postmenopausal bone loss in the femoral neck is prevented by conjugated estrogens, 1alpha(OH)D3, or both, whereas bone loss in the spine is not prevented by 1alpha(OH)D3. Estrogen proves effective in preventing early postmenopausal bone loss by markedly inhibiting bone turnover. Moreover, a synergistic bone-sparing effect can be expected when estrogen is administered concomitantly with 1alpha(OH)D3 rather than when used alone. PMID- 10369729 TI - Bone mineral density of the spine and femur in healthy Saudi females: relation to vitamin D status, pregnancy, and lactation. AB - Bone mineral density (BMD) measurements of the anterio-posterior lumbar spine and the proximal femur using dual-energy x-ray absorptiometry, as well as relevant clinical and biochemical parameters, were determined in 321 healthy Saudi females in order to establish reference values and to study the effects of physical and lifestyle factors on BMD. Mean +/- SD of age, body mass index (BMI), number of pregnancies, and total duration of lactation were 35.4 +/- 11.3 years, 26.5 +/- 5.2 kg/m2, 3.1 +/- 3.1, and 23.7 +/- 42.4 months, respectively. Mean +/- SD of serum calcium, 25-hydroxyvitamin D (25OHD), and PTH levels were 2.37 +/- 0.09 mmol/liter, 24.5 +/- 17.2 nmol/liter, and 52.0 +/- 30.8 pg/ml, respectively. Peak BMD values were observed around age 35 years at the spine and earlier at the femur. Compared with USA females, Saudi females had lower weight-matched Z scores at the spine (-0.126 +/- 1. 078, P = 0.04), femoral neck (-0.234 +/- 0.846, P < 0.0001), and Ward's triangle (-0.269 +/- 1.015, P < 0.0001). Further, the prevalence of osteopenia and osteoporosis in subjects >/=31 years old were 18-41% and 0-7%, respectively, depending on the site examined. Severe hypovitaminosis D (25OHD level T transition at nucleotide 9168 of COL1A1 genomic sequence (GenBank AF017178) was found in three unrelated patients, during the search for OI causal mutations. This polymorphic variant was then tested in the general population by Taq I restriction of genomic DNA amplified with a specifically designed restriction enzyme site-generating oligonucleotide primer. Allelic frequencies were found to be: 0.88 (C allele) and 0.12 (T allele), respectively. PMID- 10369752 TI - Polymorphisms in the human autosomal dominant polycystic kidney disease 2 (PKD2) gene. AB - Three polymorphisms of the PKD2 (MIM 173910) gene in patients with autosomal dominant polycystic kidney disease are reported: (1) a substitution from ATT (isoleucine) to GTT (valine) at codon 452; (2) a substitution from CGG (arginine) to CAG (glutamine) at codon 848; and (3) a substitution from G to A in intron 4 of the gene. The minor allelic frequencies of codon 452 and intron 4 in the Korean population were estimated to be 0.03 and 0.32, respectively. Although the codon 848 substitution was not observed in 45 unrelated healthy Korean people, the substitution did not cosegregate with the disease phenotype, suggesting that this was a rare, non-deleterious alteration. PMID- 10369753 TI - Archaeal nucleosome positioning sequence from Methanothermus fervidus. AB - DNA in Methanothermus fervidus, a hyperthermophilic archaeon, is constrained into archaeal nucleosomes in vivo by the archaeal histones HMfA and HMfB. Here, we document the translational and rotational positioning of archaeal nucleosome assembly in vitro by a sequence from the 7S RNA encoding region of the M. fervidus genome. The minor groove of the DNA at the center of the DNA sequence, protected from micrococcal nuclease digestion by incorporation into a positioned archaeal nucleosome, faces away from the archaeal histone core. PMID- 10369754 TI - Dynamic DNA contacts observed in the NMR structure of winged helix protein-DNA complex. AB - Genesis is an HNF-3/fkh homologous protein. By using multi-dimensional NMR techniques, we have obtained the solution structure and backbone dynamics of Genesis complexed with a 17 base-pair DNA. Our results indicate that both the local folding and dynamic properties of Genesis are perturbed when it binds to the DNA site. Our data show that a conserved flexible amino acid sequence (wing 1) makes dynamic contacts to DNA in the complex and a short helix is induced by Genesis-DNA interactions. Our data indicate that, unlike the HNF-3gamma/DNA complex, a magnesium ion is not required in forming the stable Genesis-DNA complex. PMID- 10369755 TI - Polyanionic inhibitors of phosphoglycerate mutase: combined structural and biochemical analysis. AB - The effects that the inhibitors inositol hexakisphosphate and benzene tri-, tetra and hexacarboxylates have on the phosphoglycerate mutases from Saccharomyces cerevisiae and Schizosaccharomyces pombe have been determined. Their Kivalues have been calculated, and the ability of the inhibitors to protect the enzymes against limited proteolysis investigated. These biochemical data have been placed in a structural context by the solution of the crystal structures of S. cerevisiae phosphoglycerate mutase soaked with inositol hexakisphosphate or benzene hexacarboxylate. These large polyanionic compounds bind to the enzyme so as to block the entrance to the active-site cleft. They form multiple interactions with the enzyme, consistent with their low Kivalues, and afford good protection against limited proteolysis of the C-terminal region by thermolysin. The inositol compound is more efficacious because of its greater number of negative charges. The S. pombe phosphoglycerate mutase that is inherently lacking a comparable C-terminal region has higher Kivalues for the compounds tested. Moreover, the S. pombe enzyme is less sensititive to proteolysis, and the presence or absence of the inhibitor molecules has little effect on susceptibility to proteolysis. PMID- 10369756 TI - Kinetic and thermodynamic stabilization of the betagamma-crystallin homolog spherulin 3a from Physarum polycephalum by calcium binding. AB - Globular proteins may be stabilized, either intrinsically, at the various levels of the structural hierarchy, or extrinsically, by ligand binding. In the case of the dormant all-beta protein spherulin 3a (S3a) from the slime mold Physarum polycephalum, binding of calcium ions causes extreme kinetic and thermodynamic stabilization. S3a is the only known single-domain member of the two Greek key superfamily of betagamma-crystallins sharing the extreme long-term stability of its homologs in vertebrate eye lens. Spectral analysis allows two Ca2+-binding sites with KD=9 microM and 200 microM to be distinguished. Unfolding in the absence and in the presence of Ca2+gives evidence for extreme kinetic stabilization of the protein: In the absence of Ca2+, the half-time of unfolding in 2. 5 M guanidinium chloride (GdmCl) equals 8.3 minutes, whereas in the presence of Ca2+, even in 7.5 M GdmCl, it exceeds nine hours. To reach the equilibrium of unfolding in the absence and in the presence of Ca2+takes one day and eight weeks, respectively. The corresponding Gibbs free energies (based on the two-state model) are 77 and 135 kJ/mol. Saturation of S3a with Ca2+leads to an upward shift of the temperature-induced equilibrium transition by ca 20 deg. C. The in situ Ca2+concentration in the spherules is sufficient for the complete complexation of S3a in vivo. PMID- 10369757 TI - Probing activation of the prokaryotic arginine transcriptional regulator using chimeric proteins. AB - The major transcription factors controlling arginine metabolism in Escherichia coli and Bacillus subtilis, ArgR and AhrC, respectively, are homologous multimeric proteins that form l -arginine-dependent DNA-binding complexes capable of repressing transcription of the biosynthetic genes (both), activating transcription of catabolic genes (AhrC only) or facilitating plasmid dimer resolution (both). Multimerisation and l -arginine binding are associated with the C-terminal 70-80 residues; the N-terminal regions contain a winged helix-turn helix DNA-binding domain. We have constructed chimeric genes in which the sequences for the N and C-terminal domains have been swapped. The resultant chimeric proteins and their corresponding native proteins have been analysed for their ability to multimerise and bind DNA operator sites in an L-arginine dependent fashion. Gel filtration and equilibrium sedimentation analysis are consistent with the formation of hexamers by all four proteins in the presence of L-arginine and at high protein concentrations (>100 nM monomer). The hexamer sedimentation coefficients suggest that there is a reduction in molecular volume upon binding L-arginine, consistent with a conformational change accompanying an allosteric activation of DNA-binding. In the absence of L-arginine or at lower protein concentrations, the hexamers are clearly in rapid equilibrium with smaller subunits, whose dominant species appear to be based on trimers, as expected from the crystal structure of the ArgR C-terminal fragment, with the exception of the ArgR-C chimera, which apparently dissociates into dimers, suggesting that in the intact protein the DNA-binding domains may have a significant dimeric interaction. The hexamer-trimer Kdis in the micromolar range, suggesting that trimers are the principal species at in vivo concentrations.DNA binding by all four proteins has been probed by gel retardation and DNase I footprinting analysis using all three types of naturally occurring operators: biosynthetic sites encompassing two 18 bp ARG boxes separated by 2 bp; biosynthetic sites containing two such boxes and a third 18 bp ARG box at a distance of 100 bp downstream, i.e. within the structural gene; and finally a catabolic operator which contains a single ARG box site. The data show that all four proteins bind to the operators at the expected regions in an L-arginine dependent fashion. From the apparent affinities of the chimeras for each target site, there is no obvious sequence-specificity associated with the N-terminal domains; rather the data can be interpreted in terms of differential allosteric activation, including DNA binding in the absence of L-arginine.Remarkably, the proteins show apparent "anti-competition" in the presence of excess, specific DNA fragments in gel retardation. This appears to be due to assembly of an activated form of the protein, probably hexamers, on the operator DNA. The data are discussed in terms of the current models for the mode of action of both native proteins. PMID- 10369758 TI - Eukaryotic signalling domain homologues in archaea and bacteria. Ancient ancestry and horizontal gene transfer. AB - Phyletic distributions of eukaryotic signalling domains were studied using recently developed sensitive methods for protein sequence analysis, with an emphasis on the detection and accurate enumeration of homologues in bacteria and archaea. A major difference was found between the distributions of enzyme families that are typically found in all three divisions of cellular life and non enzymatic domain families that are usually eukaryote-specific. Previously undetected bacterial homologues were identified for# plant pathogenesis-related proteins, Pad1, von Willebrand factor type A, src homology 3 and YWTD repeat containing domains. Comparisons of the domain distributions in eukaryotes and prokaryotes enabled distinctions to be made between the domains originating prior to the last common ancestor of all known life forms and those apparently originating as consequences of horizontal gene transfer events. A number of transfers of signalling domains from eukaryotes to bacteria were confidently identified, in contrast to only a single case of apparent transfer from eukaryotes to archaea. PMID- 10369759 TI - The topological mechanism of phage lambda integrase. AB - Bacteriophage lambda integrase (Int) is a versatile site-specific recombinase. In concert with other proteins, it mediates phage integration into and excision out of the bacterial chromosome. Int recombines intramolecular sites in inverse or direct orientation or sites on separate DNA molecules. This wide spectrum of Int mediated reactions has, however, hindered our understanding of the topology of Int recombination. By systematically analyzing the topology of Int reaction products and using a mathematical method called tangles, we deduce a unified model for Int recombination. We find that, even in the absence of (-) supercoiling, all Int reactions are chiral, producing one of two possible enantiomers of each product. We propose that this chirality reflects a right handed DNA crossing within or between recombination sites in the synaptic complex that favors formation of right-handed Holliday junction intermediates. We demonstrate that the change in linking number associated with excisive inversion with relaxed DNA is equally +2 and -2, reflecting two different substrates with different topology but the same chirality. Additionally, we deduce that integrative Int recombination differs from excisive recombination only by additional plectonemic (-) DNA crossings in the synaptic complex: two with supercoiled substrates and one with relaxed substrates. The generality of our results is indicated by our finding that two other members of the integrase superfamily of recombinases, Flp of yeast and Cre of phage P1, show the same intrinsic chirality as lambda Int. PMID- 10369760 TI - Supercoiling-dependent site-specific binding of HU to naked Mu DNA. AB - Using HU chemical nucleases to probe HU-DNA interactions, we report here for the first time site-specific binding of HU to naked DNA. An unique feature of this interaction is the absolute requirement for negative DNA supercoiling for detectable levels of site-specific DNA binding. The HU binding site is the Mu spacer between the L1 and L2 transposase binding sites. Our results suggest recognition of an altered DNA structure which is induced by DNA supercoiling. We propose that recruitment of HU to this naked DNA site induces the DNA bending required for productive synapsis and transpososome assembly. Implications of HU as a supercoiling sensor with a potential in vivo regulatory role are discussed. Finally, using HU nucleases we have also shown that non-specific DNA binding by HU is stimulated by increasing levels of supercoiling. PMID- 10369761 TI - Specificity from the synapsis of DNA elements by the Sfi I endonuclease. AB - The synapsis of DNA sites is a prerequisite for the reactions of many proteins that act at specific DNA sequences. The requirement for synapsis was investigated by analysing the reactions of Sfi I, a tetrameric restriction enzyme that cleaves DNA only after interacting with two recognition sites. In the presence of Mg2+, oligonucleotide duplexes with the cognate recognition sequence were cleaved rapidly, with cooperative kinetics, while non-cognate duplexes were not cleaved. In the absence of Mg2+, the primary complex formed by Sfi I with cognate DNA contained two duplexes synapsed by the tetramer: a secondary complex containing one duplex was seen only at elevated Sfi I concentrations. In contrast, the principal complex with non-cognate DNA contained one duplex bound to Sfi I. Pairs of non-cognate duplexes, or one cognate and one non-cognate duplex, generally failed to form synaptic complexes. On adding Mg2+to complexes with cognate DNA, cleavage occurred much more rapidly in the synaptic complex than in the secondary complex. DNA synapsis thus acts to enhance the specificity of Sfi I for its recognition sequence, by demanding two cognate sites for a catalytically active complex and by excluding non-cognate sites from the synaptic complex. PMID- 10369762 TI - Global structure and flexibility of hairpin ribozymes with extended terminal helices. AB - Global structure and flexibility of three different hairpin ribozyme constructs have been analyzed by measuring their electric dichroism decay in various buffers at temperatures between 2 and 30 degrees C. The hairpin ribozyme is characterized by two independently folding domains A and B that are connected through a hinge and have to interact to enable catalysis. The analyzed constructs feature extended terminal helices 1 and 4 with 27 and 25 bp, respectively, to increase the sensitivity of the molecular rotational diffusion time constants with respect to the interdomain bending angle. Constructs HP1 and HP2 cannot cleave because of a G+1A change at the 3'-side of the cleavage site; in HP1 the helices 2 and 3 that flank the hinge form a continuous double helical segment; in HP2 and HP3, a six nucleotide bulge confers flexibility to the expected bending site; HP3 is a cleavable form of HP2 with a G+1-base. For comparison, a standard RNA double helix with 72 bp was included in our analysis. The dichroism decay curves of the hairpin constructs after pulses of low electric field strengths can be fitted to single exponentials taus, whereas the curves after pulses of high field strengths require two exponentials. In all cases, time constants increase with RNA concentration, indicating intermolecular interactions. Extrapolation of the tausvalues measured in standard buffer (50 mM Tris (pH 7.5) and 12 mM MgCl2) to zero RNA concentration provide values of 112, 93, and 73 ns for HP1, HP2 and HP3, respectively, at 30 degrees C, indicating increasingly compact structures. The 72 bp RNA reference under corresponding conditions did not show a dependence of its decay time constant on the RNA concentration nor on the field strength; its time constant is 175 ns (standard buffer, 30 degrees C). The observation of two relaxation processes for the hairpin constructs at high field strengths indicates stretching to a more elongated state; the fast process with a time constant of the order of 50 ns is assigned to reversion of stretching, the slow process to overall rotation. The overall rotational time of the stretched state at 20 degrees C is close to that for a completely stretched rigid state; at 30 degrees C the experimental values are around 70 % of that expected for a completely stretched rigid state, indicating flexibility and/or residual bending. Bead models were constructed to simulate dichroism decay curves. The time constants observed for the 72 bp RNA are as expected for a rigid rod with a rise of 2.8 A per base-pair. Based on this rise per base-pair for models of a V and a Y-shape, we estimate average bending angles of 80(+/-20) degrees and 105 (+/-25) degrees, respectively, for the catalytically active hairpin ribozyme HP3. The energy required for stretching is of the order of the thermal energy. PMID- 10369763 TI - Inhibition of Escherichia coli RNA polymerase by bacteriophage T7 gene 2 protein. AB - The 64 amino acid residue product of bacteriophage T7 gene 2 (gp2) binds the Escherichia coli RNA polymerase and inhibits transcription. We localized the gp2 binding site to within 53 amino acid residues in the functionally dispensable region of the RNA polymerase beta' subunit. We investigated the effect of gp2 on transcription at a -10/-35 promoter and at an "extended -10" promoter. Our results indicate that binding of gp2 to the sigma70holoenzyme (Esigma70) prevents promoter recognition at -10/-35 promoters. Once open promoter complexes are formed, however, Esigma70transcription is resistant to gp2, since gp2 can no longer bind RNA polymerase. Surprisingly, transcription inhibition by gp2 is both sigma and promoter-specific. gp2 has little effect on Esigma70transcription from an extended -10 promoter, which does not depend on sigma70region 4 interactions with the -35 promoter box for its activity. gp55-dependent phage T4 late promoter transcription is also resistant to gp2. From these results, we conclude that the interaction of the sigma70region 4 with the -35 consensus promoter element is the primary target of gp2 inhibition. PMID- 10369764 TI - Erythromycin resistance mutations in ribosomal proteins L22 and L4 perturb the higher order structure of 23 S ribosomal RNA. AB - We have used chemical modification to examine the conformation of 23 S rRNA in Escherichia coli ribosomes bearing erythromycin resistance mutations in ribosomal proteins L22 and L4. Changes in reactivity to chemical probes were observed at several nucleotide positions scattered throughout 23 S rRNA. The L4 mutation affects the reactivity of G799 and U1255 in domain II and that of A2572 in domain V. The L22 mutation influences modification in domain II at positions m5U747, G748, and A1268, as well as at A1614 in domain III and G2351 in domain V. The reactivity of A789 is weakly enhanced by both the L22 and L4 mutations. None of these nucleotide positions has previously been associated with macrolide antibiotic resistance. Interestingly, neither of the ribosomal protein mutations produces any detectable effects at or within the vicinity of A2058 in domain V, the site most frequently shown to confer macrolide resistance when altered by methylation or mutation. Thus, while L22 and L4 bind primarily to domain I of 23 S rRNA, erythromycin resistance mutations in these ribosomal proteins perturb the conformation of residues in domains II, III and V and affect the action of antibiotics known to interact with nucleotide residues in the peptidyl transferase center of domain V. These results support the hypothesis that ribosomal proteins interact with rRNA at multiple sites to establish its functionally active three-dimensional structure, and suggest that these antibiotic resistance mutations act by perturbing the conformation of rRNA. PMID- 10369766 TI - Effect of magnesium on cruciform extrusion in supercoiled DNA. AB - Recently, it was reported that Mg2+greatly facilitates cruciform extrusion in the short palindromes of supercoiled DNA, thereby allowing the formation of cruciform structures in vivo. Because of the potential biological importance of this phenomenon, we undertook a broader study of the effect of Mg2+on a cruciform extrusion in supercoiled DNA. The method of two-dimensional gel electrophoresis was used to detect the cruciform extrusion both in the absence and in the presence of these ions. Our results show that Mg2+shifts the cruciform extrusion in the d(CCC(AT)16GGG) palindrome to a higher, rather than to a lower level of supercoiling. In order to study possible sequence-specific properties of the short palindromes for which the unusual cruciform extrusion in the presence Mg2+was reported, we constructed a plasmid with a longer palindromic region. This region bears the same sequences in the hairpin loops and four-arm junction as the short palindrome, except that the short stems of the hairpins are extended. The extension allowed us to overcome the limitation of our experimental approach which cannot be used for very short palindromes. Our results show that Mg2+also shifts the cruciform extrusion in this palindrome to a higher level of supercoiling. These data suggest that cruciform extrusion in the short palindromes at low supercoiling is highly improbable. We performed a thermodynamic analysis of the effect of Mg2+on cruciform extrusion. The treatment accounted for the effect of Mg2+on both free energy of supercoiling and the free energy of cruciform structure per se. Our analysis showed that although the level of supercoiling required for the cruciform extrusion is not reduced by Mg2+, the ions reduce the free energy of the cruciform structure. PMID- 10369765 TI - DNA replication errors produced by the replicative apparatus of Escherichia coli. AB - It has been hard to detect forward mutations generated during DNA synthesis in vitro by replicative DNA polymerases, because of their extremely high fidelity and a high background level of pre-existing mutations in the single-stranded template DNA used. Using the oriC plasmid DNA replication in vitro system and the rpsL forward mutation assay, we examined the fidelity of DNA replication catalyzed by the replicative apparatus of Escherichia coli. Upon DNA synthesis by the fully reconstituted system, the frequency of rpsL-mutations in the product DNA was increased to 1.9x10(-4), 50-fold higher than the background level of the template DNA. Among the mutations generated in vitro, single-base frameshifts predominated and occurred with a pattern similar to those induced in mismatch repair deficient E. coli cells, indicating that the major replication error was slippage at runs of the same nucleotide. Large deletions and other structural alterations of DNA appeared to be induced also during the action of the replicative apparatus. PMID- 10369767 TI - Origin and phylogenetic distribution of Alu DNA repeats: irreversible events in the evolution of primates. AB - Over the past 60 million years, or so, approximately one million copies of Alu DNA repeats have accumulated in the genome of primates, in what appears to be an ongoing process. We determined the phylogenetic distribution of specific Alu (and other) DNA repeats in the genome of several primates: human, chimpanzee, gorilla, orangutan, baboon, rhesus, and macaque. At the population level studied, the majority of the repeats was found to be fixed in the primate species. Our data suggest that new Alu elements arise in unique, irreversible events, in a mechanism that seems to preclude precise excision and loss. The same insertions did not arise independently in two species. Once inserted and genetically fixed, the DNA elements are retained in all descendant lineages. The irreversible expansion of Alu s introduces a vector of time into the evolutionary process, and provides realistic (rather than statistical) answers to questions on phylogenies. In contrast to point mutations, the present distribution of individual Alu s is congruent with just one phylogeny. We submit that only irreversible and taxonomically relevant events are at the molecular basis of evolution. Most point mutations do not belong to this category. PMID- 10369768 TI - Topology of Xer recombination on catenanes produced by lambda integrase. AB - Xer site-specific recombination at the psi site from plasmid pSC101 displays topological selectivity, such that recombination normally occurs only between directly repeated sites on the same circular DNA molecule. This intramolecular selectivity is important for the biological role of psi, and is imposed by accessory proteins PepA and ArcA acting at accessory DNA sequences adjacent to the core recombination site. Here we show that the selectivity for intramolecular recombination at psi can be bypassed in multiply interlinked catenanes. Xer site specific recombination occurred relatively efficiently between antiparallel psi sites located on separate rings of right-handed torus catenanes containing six or more nodes. This recombination introduced one additional node into the catenanes. Antiparallel sites on four-noded right-handed catenanes, the normal product of Xer recombination at psi, were not recombined efficiently. Furthermore, parallel psi sites on right-handed torus catenanes were not substrates for Xer recombination. These findings support a model in which psi sites are plectonemically interwrapped, trapping a precise number of supercoils that are converted to four catenation nodes by Xer strand exchange. PMID- 10369769 TI - Mutational analysis of Saccharomyces cerevisiae Smf1p, a member of the Nramp family of metal transporters. AB - We have recently shown that a member of the Nramp family of metal transporters, Saccharomyces cerevisiae Smf1p, is tightly regulated at the level of protein stability and protein sorting. Under metal replete conditions, Smf1p is targeted to the vacuole for degradation in a manner dependent on the S. cerevisiaeBSD2 gene product, but under metal starvation conditions, Smf1p accumulates at the cell surface. Here, we have addressed whether Smf1p activity may be necessary for its regulation by metal ions and Bsd2p. Well conserved residues within transmembrane domain 4 and the transport signature sequence of Smf1p were mutagenized. We identified two mutants, G190A and G424A, which destroyed Smf1p activity as monitored by complementation of a smf1 mutation. Notably, these mutations also abolished control by metal ions and Bsd2p, suggesting that Smf1p metal transport function may be necessary for its regulation. Two additional mutants isolated (Q419A and E423A) exhibited wild-type complementation activity and were properly targeted for vacuolar degradation in a Bsd2-dependent manner. However, these mutants failed to re-distribute to the plasma membrane under conditions of metal starvation. A model is proposed herein describing the probable role of Smf1 protein conformation in directing its movement to the vacuole versus cell surface in response to changes in metal ion availability. PMID- 10369770 TI - Developmental regulation of the aldolase A muscle-specific promoter during in vivo muscle maturation is controlled by a nuclear receptor binding element. AB - During the post-natal period, skeletal muscles undergo important modifications leading to the appearance of different types of myofibers which exhibit distinct contractile and metabolic properties. This maturation process results from the activation of the expression of different sets of contractile proteins and metabolic enzymes, which are specific to the different types of myofibers. The muscle-specific promoter of the aldolase A gene (pM) is expressed mainly in fast twitch glycolytic fibers in adult body muscles. We investigate here how pM is regulated during the post-natal development of different types of skeletal muscles (slow or fast-twitch muscles, head or body muscles). We show that pM is expressed preferentially in prospective fast-twitch muscles soon after birth; pM is up-regulated specifically in body muscles only later in development. This activation pattern is mimicked by a transgene which comprises only the 355 most proximal sequences of pM. Within this region, we identify a DNA element which is required for the up-regulation of the transgene during post-natal development in body muscles. Comparison of nuclear M1-binding proteins from young or adult body muscles show no qualitative differences. Distinct M1-binding proteins are present in both young and adult tongue nuclear extracts, compared to that present in gastrocnemius extracts. PMID- 10369771 TI - Identification of a discrete intermediate in the assembly/disassembly of physalis mottle tymovirus through mutational analysis. AB - Assembly intermediates of icosahedral viruses are usually transient and are difficult to identify. In the present investigation, site-specific and deletion mutants of the coat protein gene of physalis mottle tymovirus (PhMV) were used to delineate the role of specific amino acid residues in the assembly of the virus and to identify intermediates in this process. N-terminal 30, 34, 35 and 39 amino acid deletion and single C-terminal (N188) deletion mutant proteins of PhMV were expressed in Escherichia coli. Site-specific mutants H69A, C75A, W96A, D144N, D144N-T151A, K143E and N188A were also constructed and expressed. The mutant protein lacking 30 amino acid residues from the N terminus self-assembled to T=3 particles in vivo while deletions of 34, 35 and 39 amino acid residues resulted in the mutant proteins that were insoluble. Interestingly, the coat protein (pR PhCP) expressed using pRSET B vector with an additional 41 amino acid residues at the N terminus also assembled into T=3 particles that were more compact and had a smaller diameter. These results demonstrate that the amino-terminal segment is flexible and either the deletion or addition of amino acid residues at the N terminus does not affect T=3 capsid assembly. In contrast, the deletion of even a single residue from the C terminus (PhN188Delta1) resulted in capsids that were unstable. These capsids disassembled to a discrete intermediate with a sedimentation coefficent of 19.4 S. However, the replacement of C-terminal asparagine 188 by alanine led to the formation of stable capsids. The C75A and D144N mutant proteins also assembled into capsids that were as stable as the pR PhCP, suggesting that C75 and D144 are not crucial for the T=3 capsid assembly. pR PhW96A and pR PhD144N-T151A mutant proteins failed to form capsids and were present as heterogeneous aggregates. Interestingly, the pR PhK143E mutant protein behaved in a manner similar to the C-terminal deletion protein in forming unstable capsids. The intermediate with an s value of 19.4 S was the major assembly product of pR PhH69A mutant protein and could correspond to a 30mer. It is possible that the assembly or disassembly is arrested at a similar stage in pR PhN188Delta1, pR PhH69A and pR PhK143E mutant proteins. PMID- 10369772 TI - Three-dimensional structure of physalis mottle virus: implications for the viral assembly. AB - The structure of the T=3 single stranded RNA tymovirus, physalis mottle virus (PhMV), has been determined to 3.8 A resolution. PhMV crystals belong to the rhombohedral space group R 3, with one icosahedral particle in the unit cell leading to 20-fold non-crystallographic redundancy. Polyalanine coordinates of the related turnip yellow mosaic virus (TYMV) with which PhMV coat protein shares 32 % amino acid sequence identity were used for obtaining the initial phases. Extensive phase refinement by real space molecular replacement density averaging resulted in an electron density map that revealed density for most of the side chains and for the 17 residues ordered in PhMV, but not seen in TYMV, at the N terminus of the A subunits. The core secondary and tertiary structures of the subunits have a topology consistent with the capsid proteins of other T=3 plant viruses. The N-terminal arms of the A subunits, which constitute 12 pentamers at the icosahedral 5-fold axes, have a conformation very different from the conformations observed in B and C subunits that constitute hexameric capsomers with near 6-fold symmetry at the icosahedral 3-fold axes. An analysis of the interfacial contacts between protein subunits indicates that the hexamers are held more strongly than pentamers and hexamer-hexamer contacts are more extensive than pentamer-hexamer contacts. These observations suggest a plausible mechanism for the formation of empty capsids, which might be initiated by a change in the conformation of the N-terminal arm of the A subunits. The structure also provides insights into immunological and mutagenesis results. Comparison of PhMV with the sobemovirus, sesbania mosaic virus reveals striking similarities in the overall tertiary fold of the coat protein although the capsid morphologies of these two viruses are very different. PMID- 10369773 TI - RNA secondary structure prediction based on free energy and phylogenetic analysis. AB - We describe a computational method for the prediction of RNA secondary structure that uses a combination of free energy and comparative sequence analysis strategies. Using a homology-based sequence alignment as a starting point, all favorable pairings with respect to the Turner energy function are identified. Each potentially paired region within a multiple sequence alignment is scored using a function that combines both predicted free energy and sequence covariation with optimized weightings. High scoring regions are ranked and sequentially incorporated to define a growing secondary structure. Using a single set of optimized parameters, it is possible to accurately predict the foldings of several test RNAs defined previously by extensive phylogenetic and experimental data (including tRNA, 5 S rRNA, SRP RNA, tmRNA, and 16 S rRNA). The algorithm correctly predicts approximately 80% of the secondary structure. A range of parameters have been tested to define the minimal sequence information content required to accurately predict secondary structure and to assess the importance of individual terms in the prediction scheme. This analysis indicates that prediction accuracy most strongly depends upon covariational information and only weakly on the energetic terms. However, relatively few sequences prove sufficient to provide the covariational information required for an accurate prediction. Secondary structures can be accurately defined by alignments with as few as five sequences and predictions improve only moderately with the inclusion of additional sequences. PMID- 10369774 TI - High precision solution structure of the C-terminal KH domain of heterogeneous nuclear ribonucleoprotein K, a c-myc transcription factor. AB - Among it's many reported functions, heterogeneous nuclear ribonucleoprotein (hnRNP) K is a transcription factor for the c- myc gene, a proto-oncogene critical for the regulation of cell growth and differentiation. We have determined the solution structure of the Gly26-->Arg mutant of the C-terminal K homology (KH) domain of hnRNP K by NMR spectroscopy. This is the first structure investigation of hnRNP K. Backbone residual dipolar couplings, which provide information that is fundamentally different from the standard NOE-derived distance restraints, were employed to improve structure quality. An independent assessment of structure quality was achieved by comparing the backbone15N T1/T2ratios to the calculated structures. The C-terminal KH module of hnRNP K (KH3) is revealed to be a three-stranded beta-sheet stacked against three alpha helices, two of which are nearly parallel to the strands of the beta-sheet. The Gly26-->Arg mutation abolishes single-stranded DNA binding without altering the overall fold of the protein. This provides a clue to possible nucleotide binding sites of KH3. It appears unlikely that the solvent-exposed side of the beta-sheet will be the site of protein-nucleic acid complex formation. This is in contrast to the earlier theme for protein-RNA complexes incorporating proteins structurally similar to KH3. We propose that the surface of KH3 that interacts with nucleic acid is comparable to the region of DNA interaction for the double stranded DNA-binding domain of bovine papillomavirus-1 E2 that has a three dimensional fold similar to that of KH3. PMID- 10369775 TI - Conformational dynamics and molecular recognition: backbone dynamics of the estrogen receptor DNA-binding domain. AB - We examined the internal mobility of the estrogen receptor DNA-binding domain (ER DBD) using NMR15N relaxation measurements and compared it to that of the glucocorticoid receptor DNA-binding domain (GR DBD). The studied protein fragments consist of residues Arg183-His267 of the human ER and residues Lys438 Gln520 of the rat GR. The15N longitudinal (R1) and transverse (R2) relaxation rates and steady state {1H}-15N nuclear Overhauser enhancements (NOEs) were measured at 30 degrees C at1H NMR frequencies of 500 and 600 MHz. The NOE versus sequence profile and calculated order parameters for ER DBD backbone motions indicate enhanced internal dynamics on pico- to nanosecond time-scales in two regions of the core DBD. These are the extended strand which links the DNA recognition helix to the second zinc domain and the larger loop region of the second zinc domain. The mobility of the corresponding regions of the GR DBD, in particular that of the second zinc domain, is more limited. In addition, we find large differences between the ER and GR DBDs in the extent of conformational exchange mobility on micro- to millisecond time-scales. Based on measurements of R2as a function of the15N refocusing (CPMG) delay and quantitative (Lipari-Szabo type) analysis, we conclude that conformational exchange occurs in the loop of the first zinc domain and throughout most of the second zinc domain of the ER DBD. The conformational exchange dynamics in GR DBD is less extensive and localized to two sites in the second zinc domain. The different dynamical features seen in the two proteins is consistent with previous studies of the free state structures in which the second zinc domain in the ER DBD was concluded to be disordered whereas the corresponding region of the GR DBD adopts a stable fold. Moreover, the regions of the ER DBD that undergo conformational dynamics on the micro- to millisecond time-scales in the free state are involved in intermolecular protein-DNA and protein-protein interactions in the dimeric bound state. Based on the present data and the previously published dynamical and DNA binding properties of a GR DBD triple mutant which recognize an ER binding site on DNA, we argue that the free state dynamical properties of the nuclear receptor DBDs is an important element in molecular recognition upon DNA binding. PMID- 10369776 TI - The structure of adrenodoxin reductase of mitochondrial P450 systems: electron transfer for steroid biosynthesis. AB - Adrenodoxin reductase is a monomeric 51 kDa flavoenzyme that is involved in the biosynthesis of all steroid hormones. The structure of the native bovine enzyme was determined at 2.8 A resolution, and the structure of the respective recombinant enzyme at 1.7 A resolution. Adrenodoxin reductase receives a two electron package from NADPH and converts it to two single electrons that are transferred via adrenodoxin to all mitochondrial cytochromes P 450. The structure suggests how the observed flavin semiquinone is stabilized. A striking feature is the asymmetric charge distribution, which most likely controls the approach of the electron carrier adrenodoxin. A model for the interaction is proposed. Adrenodoxin reductase shows clear sequence homology to half a dozen proteins identified in genome analysis projects, but neither sequence nor structural homology to established, functionally related electron transferases. Yet, the structure revealed a relationship to the disulfide oxidoreductases, permitting the assignment of the NADP-binding site. PMID- 10369777 TI - Structure of aspartate-beta-semialdehyde dehydrogenase from Escherichia coli, a key enzyme in the aspartate family of amino acid biosynthesis. AB - Aspartate beta-semialdehyde dehydrogenase (ASADH) lies at the first branch point in an essential aspartic biosynthetic pathway found in bacteria, fungi and the higher plants. Mutations in the asd gene encoding for ASADH that produce an inactive enzyme are lethal, which suggests that ASADH may be an effective target for antibacterial, herbicidal and fungicidal agents. We have solved the crystal structure of the Escherichia coli enzyme to 2.5 A resolution using single isomorphous replacement and 3-fold non-crystallographic symmetry. Each monomer has an N-terminal nucleotide-binding domain and a dimerisation domain. The presence of an essential cysteine locates the active site in a cleft between the two domains. The functional dimer has the appearance of a butterfly, with the NADP-binding domains forming the wings and the dimerisation domain forming the body.A histidine residue is identified as a likely acid/base catalyst in the enzymic reaction. Other amino acids implicated in the enzymic activity by mutagenesis are found in the active site region and define the substrate binding pocket. PMID- 10369779 TI - MAD structure of Pseudomonas nautica dimeric cytochrome c552 mimicks the c4 Dihemic cytochrome domain association. AB - The monohemic cytochrome c552from Pseudomonas nautica (c552-Pn) is thought to be the electron donor to cytochrome cd1, the so-called nitrite reductase (NiR). It shows as high levels of activity and affinity for the P. nautica NiR (NiR-Pn), as the Pseudomonas aeruginosa enzyme (NiR-Pa). Since cytochrome c552is by far the most abundant electron carrier in the periplasm, it is probably involved in numerous other reactions. Its sequence is related to that of the c type cytochromes, but resembles that of the dihemic c4cytochromes even more closely. The three-dimensional structure of P. nautica cytochrome c552has been solved to 2.2 A resolution using the multiple wavelength anomalous dispersion (MAD) technique, taking advantage of the presence of the eight Fe heme ions in the asymmetric unit. Density modification procedures involving 4-fold non crystallographic averaging yielded a model with an R -factor value of 17.8 % (Rfree=20.8 %). Cytochrome c552forms a tight dimer in the crystal, and the dimer interface area amounts to 19% of the total cytochrome surface area. Four tighly packed dimers form the eight molecules of the asymmetric unit. The c552dimer is superimposable on each domain of the monomeric cytochrome c4from Pseudomomas stutzeri (c4-Ps), a dihemic cytochrome, and on the dihemic c domain of flavocytochrome c of Chromatium vinosum (Fcd-Cv). The interacting residues which form the dimer are both similar in character and position, which is also true for the propionates. The dimer observed in the crystal also exists in solution. It has been hypothesised that the dihemic c4-Ps may have evolved via monohemic cytochrome c gene duplication followed by evolutionary divergence and the adjunction of a connecting linker. In this process, our dimeric c552structure might be said to constitute a "living fossile" occurring in the course of evolution between the formation of the dimer and the gene duplication and fusion. The availability of the structure of the cytochrome c552-Pn and that of NiR from P. aeruginosa made it possible to identify putative surface patches at which the docking of c552to NiR-Pn may occur. PMID- 10369778 TI - Regulatory features of the trp operon and the crystal structure of the trp RNA binding attenuation protein from Bacillus stearothermophilus. AB - Characterization of both the cis and trans -acting regulatory elements indicates that the Bacillus stearothermophilustrp operon is regulated by an attenuation mechanism similar to that which controls the trp operon in Bacillus subtilis. Secondary structure predictions indicate that the leader region of the trp mRNA is capable of folding into terminator and anti- terminator RNA structures. B. stearothermophilus also encodes an RNA-binding protein with 77% sequence identity with the RNA-binding protein (TRAP) that regulates attenuation in B. subtilis. The X-ray structure of this protein has been determined in complex with L tryptophan at 2.5 A resolution. Like the B. subtilis protein, B. stearothermophilus TRAP has 11 subunits arranged in a ring-like structure. The central cavities in these two structures have different sizes and opposite charge distributions, and packing within the B. stearothermophilus TRAP crystal form does not generate the head-to-head dimers seen in the B. subtilis protein, suggesting that neither of these properties is functionally important. However, the mode of L-tryptophan binding and the proposed RNA binding surfaces are similar, indicating that both proteins are activated by l -tryptophan and bind RNA in essentially the same way. As expected, the TRAP:RNA complex from B. stearothermophilus is significantly more thermostable than that from B. subtilis, with optimal binding occurring at 70 degrees C. PMID- 10369780 TI - Characterization of the dimer interface of transcription factor NFkappaB p50 homodimer. AB - Dimers of the Rel/NFkappaB transcription factor family form with differential stabilities through the combinatorial association of five polypeptides: p50, p52, p65, cRel, and RelB. Here, we have characterized the nature of the monomer-dimer equilibrium of the p50 homodimer. Sedimentation equilibrium studies show that the equilibrium constant for p50 dimer dissociation is in the low micromolar range. Using the X-ray crystal structure of the p50 homodimer as a guide, we have created site-directed alanine mutations at ten dimer-forming residues in p50 and measured their effects on p50 homodimerization. Characterization of these alanine mutants by a series of chemical crosslinking, size-exclusion chromatography, and sedimentation equilibrium experiments shows that the most critical residue in stabilizing the p50 dimer interface is Y267. Sedimentation equilibrium experiments show that an alanine substitution at position 267 destabilizes the dimer interface by 2.0 kcal/mol. Alanine substitutions at two other positions, L269 and V310, significantly destabilize the p50 dimer interface. These two residues are observed to mediate critical interactions in the crystal structure. Together, these three residues constitute the "hot-spot" of protein-protein interaction in p50 dimerization. Of the four charged residues in the dimer interface, R252, D254, E265, and D302, only D302 contributes significantly to p50 dimer stability. D254 appears to slightly destabilize the subunit interface. Although residues H304, R305, and F307 occupy positions at the hydrophobic core of the interface and appear to be involved in multiple interactions in the X-ray crystal structure, alanine substitutions at these positions do not significantly reduce the affinity for p50 dimerization. Upon evaluating the roles of these amino acid residues at the p50 dimer interface, we propose that differential contributions of a few key residues dictate the selectivity of dimer formation within the Rel/NFkappaB family. PMID- 10369781 TI - NMR hydrogen exchange of the OB-fold protein LysN as a function of denaturant: the most conserved elements of structure are the most stable to unfolding. AB - The structure of LysN contains an OB-fold motif composed of a structurally conserved five-stranded beta-barrel capped by a poorly conserved alpha-helix between strands beta3 and beta4. Two additional alpha-helices, unique to the LysN structure, flank the N terminus of the OB-fold. The stability of LysN to unfolding has been investigated with NMR native state hydrogen exchange measurements as a function of guanidinium hydrochloride concentration, and equilibrium unfolding transitions monitored by ellipticity at 222 nm and fluorescence at 350 nm. The spectrophotometric measurements suggest an apparent two-state unfolding transition with DeltaGu(0) approximately 6 kcal/mol and m approximately 3 kcal/(molM). By contrast, NMR hydrogen exchange measurements manifest a distribution of DeltaGu(0) and m values which indicate that the protein can undergo subglobal unfolding. The largest DeltaGu(0) values from hydrogen exchange are for residues in the beta-sheet of the protein. These values, which reflect complete unfolding of the protein, are between 3 and 4 kcal/mol higher than those obtained from circular dichroism or fluorescence. This discrepancy may be due to the comparison of NMR hydrogen exchange parameters measured at residue-level resolution, with spectrophotometric parameters that reflect an unresolved super position of unfolding transitions of the alpha helices and beta-strands. The largest DeltaGu(0) values obtained from hydrogen exchange for the subset of residues in the alpha-helices of the protein, agree with the DeltaGu(0) values obtained from circular dichroism or fluorescence. Based on the hydrogen exchange data, however, the three alpha-helices of LysN are on average 3 kcal/mol less stable than the beta-sheet. Consistent with the subglobal unfolding of LysN evinced by hydrogen exchange, a deletion mutant that lacks the first alpha-helix of the protein retains a cooperatively folded structure. Taken together with previous results on the OB-fold proteins SN and CspA, the present results for LysN suggest that the most conserved elements of structure in the OB-fold motif are the most resistant to denaturation. In all three proteins, stability to denaturation correlates with sequence hydrophobicity. PMID- 10369782 TI - Independent nucleation and heterogeneous assembly of structure during folding of equine lysozyme. AB - The refolding of equine lysozyme from guanidinium chloride has been studied using hydrogen exchange pulse labelling in conjunction with NMR spectroscopy and stopped flow optical methods. The stopped flow optical experiments indicate that extensive hydrophobic collapse occurs rapidly after the initiation of refolding. Pulse labelling experiments monitoring nearly 50 sites within the protein have enabled the subsequent formation of native-like structure to be followed in considerable detail. They reveal that an intermediate having persistent structure within three of the four helices of the alpha-domain of the protein is formed for the whole population of molecules within 4 ms. Subsequent to this event, however, the hydrogen exchange protection kinetics are complex and highly heterogeneous. Analysis of the results by fitting to stretched exponential functions shows that a series of other intermediates is formed as a consequence of the stepwise assembly of independently nucleated local regions of structure. In some molecules the next step in folding involves the stabilisation of the remaining helix in the alpha-domain, whilst in others persistent structure begins to form in the beta domain. The formation of native-like structure throughout the beta-domain is itself heterogeneous, involving at least three kinetically distinguishable steps. Residues in loop regions throughout the protein attain persistent structure more slowly than regions of secondary structure. There is in addition evidence for locally misfolded regions of structure that reorganise on much longer timescales. The results reveal that the native state of the protein is generated by the heterogeneous assembly of a series of locally cooperative regions of structure. This observation has many features in common with the findings of recent theoretical simulations of protein folding. PMID- 10369783 TI - Catalysis, commitment and encapsulation during GroE-mediated folding. AB - The Escherichia coli GroE chaperones assist protein folding under conditions where no spontaneous folding occurs. To achieve this, the cooperation of GroEL and GroES, the two protein components of the chaperone system, is an essential requirement. While in many cases GroE simply suppresses unspecific aggregation of non-native proteins by encapsulation, there are examples where folding is accelerated by GroE. Using maltose-binding protein (MBP) as a substrate for GroE, it had been possible to define basic requirements for catalysis of folding. Here, we have analyzed key steps in the interaction of GroE and the MBP mutant Y283D during catalyzed folding. In addition to high temperature, high ionic strength was shown to be a restrictive condition for MBP Y283D folding. In both cases, the complete GroE system (GroEL, GroES and ATP) compensates the deceleration of MBP Y283D folding. Combining kinetic folding experiments and electron microscopy of GroE particles, we demonstrate that at elevated temperatures, symmetrical GroE particles with GroES bound to both ends of the GroEL cylinder play an important role in the efficient catalysis of MBP Y283D refolding. In principle, MBP Y283D folding can be catalyzed during one encapsulation cycle. However, because the commitment to reach the native state is low after only one cycle of ATP hydrolysis, several interaction cycles are required for catalyzed folding. PMID- 10369784 TI - SuperStar: a knowledge-based approach for identifying interaction sites in proteins. AB - An empirical method for identifying interaction sites in proteins is described and validated. The method is based entirely on experimental information about non bonded interactions occurring in small-molecule crystal structures. These data are used in the form of scatterplots that show the experimentally observed distribution of one functional group (the "contact group" or "probe") around another. A template molecule (e.g. a protein binding site) is broken down into structure fragments and the scatterplots, showing the distribution of a chosen probe around these structure fragments, are superimposed on the corresponding parts of the template. The scatterplots are then translated into a three dimensional map that shows the propensity of the probe at different positions around the template molecule. The method is illustrated for l -arabinose-binding protein, complexed with l -arabinose and with d -fucose, and for dihydrofolate reductase complexed with methotrexate. The method is validated on 122 X-ray structures of protein-ligand complexes. For all the binding sites of these proteins, propensity maps are generated for four different probes: a charged NH+3nitrogen, a carbonyl oxygen, a hydroxyl oxygen and a methyl carbon atom. Next, the maps are compared with the experimentally observed positions of ligand atoms of these types. For 74% of these ligand atoms (84% of the solvent inaccessible ones) the calculated propensity of the matching probe at the experimental positions is higher than expected by chance. For 68% of the atoms (82% of the solvent-inaccessible ones) the propensity of the matching probe is higher than that of the other three probes. These results indicate that the approach generally gives good predictions for protein-ligand interactions. The potential applications of the propensity maps range from an aid in manual docking and structure-based drug design to their use in pharmacophore development. PMID- 10369785 TI - The transferrin receptor binding site on HFE, the class I MHC-related protein mutated in hereditary hemochromatosis. AB - HFE is a class I major histocompatibility complex (MHC)-related protein that is mutated in patients with the iron storage disease hereditary hemochromatosis. HFE binds tightly to transferrin receptor (TfR), the receptor that mediates uptake of iron-loaded transferrin. The binding affinities for TfR of HFE mutants, designed using the HFE crystal structure, were measured using biosensor assays. The results allow localization of the TfR binding site on HFE to the C-terminal portion of the alpha1 domain helix and an adjacent loop, a region distinct from the ligand binding sites on class I MHC and related proteins. A biosensor-derived pH-dependent affinity profile for the HFE-TfR interaction is discussed in terms of HFE's hypothesized role in intracellular trafficking. PMID- 10369786 TI - Model of the ran-RCC1 interaction using biochemical and docking experiments. AB - RCC1, the regulator of chromosome condensation, is the guanine nucleotide exchange factor (GEF) for the nuclear Ras-like GTP-binding protein Ran. Its structure was solved by X-ray crystallography and revealed a seven-bladed beta propeller, one side of which was proposed to be the interaction site with Ran. To gain more insight into this interaction, alanine mutagenesis studies were performed on conserved residues on the surface of the structure. Purified mutant proteins were analysed by steady-state kinetic analysis of their GEF activities towards Ran. A number of residues were identified whose mutation affected either the KMor kcatof the overall reaction, or had no effect. Mutants were further analysed by plasmon surface resonance in order to get more information on individual steps of the complex reaction pathway. Ran-GDP was coupled to the sensor chip and reacted with RCC1 mutants to categorise them into different groups, demonstrating the usefulness of plasmon surface resonance in the study of complex multi-step kinetic processes. A docking solution of Ran-RCC1 structures in combination with sequence analysis allows prediction of the site of interaction between RCC1 and Ran and proposes a model for the Ran-RCC1 structure which corresponds to and extends the biochemical data. Three invariant residues which most severely affect the kcatof the reaction, D128, D182 and H304, are located in the centre of the Ran-RCC1 interface and interfere with switch II and the phosphate binding area. The structural model suggests that different guanine nucleotide exchange factors use a similar interaction site on their respective GTP-binding proteins, but that the molecular mechanisms for the release of nucleotides are likely to be different. PMID- 10369787 TI - Investigation of the interaction between DnaK and DnaJ by surface plasmon resonance spectroscopy. AB - Hsp70 chaperones assist protein folding through ATP-regulated transient association with substrates. Substrate binding by Hsp70 is controlled by DnaJ co chaperones which stimulate Hsp70 to hydrolyze ATP and, consequently, to close its substrate binding cavity allowing trapping of substrates. We analyzed the interaction of the Escherichia coli Hsp70 homologue, DnaK, with DnaJ using surface plasmon resonance (SPR) spectroscopy. Resonance signals of complex kinetic characteristics were detected when DnaK was passed over a sensor chip with coupled DnaJ. This interaction was specific as it was not detected with a functionally defective DnaJ mutant protein, DnaJ259, that carries a mutation in the HPD signature motif of the conserved J-domain. Detectable DnaK-DnaJ interaction required ATP hydrolysis by DnaK and was competitively inhibited by chaperone substrates of DnaK. For DnaK mutant proteins with amino acid substitutions in the substrate binding cavity that affect substrate binding, the strength of detected interaction with DnaJ decreased proportionally with increased strength of the substrate binding defects. These findings indicate that the detected response signals resulted from DnaJ and ATP hydrolysis-dependent association of DnaJ as substrate for DnaK. Although not considered as physiologically relevant, this association allowed us to experimentally unravel the mechanism of DnaJ action. Accordingly, DnaJ stimulates ATP hydrolysis only after association of a substrate with the substrate binding cavity of DnaK. Further analysis revealed that this coupling mechanism required the J-domain of DnaJ and was also functional for natural DnaK substrates, and thus is central to the mechanism of action of the DnaK chaperone system. PMID- 10369788 TI - Cardiovascular effects of sex hormones. PMID- 10369789 TI - Role of nitric oxide in the regulation of cardiovascular autonomic control. AB - Alteration in function of the cardiac autonomic nervous system has proved to be a powerful predictor of cardiac death or serious arrhythmia in patients with cardiac disease, yet little is known about the mechanisms by which this system is regulated. Recent evidence suggests that the gaseous molecule nitric oxide (NO) may act as an important mediator in this pathway. Histochemical staining techniques have identified neuronal populations that contain NO synthase within medullary cardio-regulatory sites and their peripheral autonomic pathways. Drugs that modulate the NO pathway (administered both systemically and into the central nervous system) cause changes in pre- and post-ganglionic sympathetic nerve activity that imply that NO serves to inhibit central sympathetic outflow. There is also evidence that NO may attenuate cardiovascular end-organ responses to sympathetic stimulation. Studies suggest that NO modulates cardiac vagal control, increasing the activity of central vagal motoneurons and, more contentiously, contributing to the bradycardic effects of vagal stimulation. NO also modulates so-called 'indirect' vagal inhibition of sympathetic cardiac responses. Additionally, central attenuation of baroreflex-mediated vagal control has been described. There is relatively little information available on the importance of NO in the regulation of human cardiovascular autonomic control. Further well controlled studies are required. PMID- 10369790 TI - Gender differences in the endothelial regulation of alpha2-adrenoceptor-mediated contraction in the rat aorta. AB - The aim of this study was to determine the possible influence of sex hormones on the contractile responses induced by clonidine, an agonist of alpha2 adrenoceptors, as well as the endothelial modulation of these responses. For this purpose, thoracic aorta segments from male (control and castrated) and female (in oestrous phase and ovariectomized) rats were used. In intact segments from the four groups of rats, clonidine (0.01-10 micromol/l) induced concentration dependent contractions, which were increased by the nitric oxide synthase inhibitor Nomega-nitro-L-arginine methyl ester (0.1 mmol/l) or by endothelium removal, but were reduced by 1 micromol/l yohimbine (an alpha2-adrenoceptor antagonist) in all animals and by 1 micromol/l indomethacin (a cyclo-oxygenase inhibitor) in control males only. The rank order of the magnitude of the maximal response was: oestrous females>ovariectomized females>control males>castrated males, whereas the sensitivity to clonidine (EC50 value) was similar in all animals. In endothelium-denuded segments, the rank order was: oestrous females=control males>ovariectomized females=castrated males. These results suggest that: (1) the presence of oestrogen or androgen increases the contraction caused by alpha2-adrenoceptor activation with clonidine; (2) endothelium negatively modulates the response to this agonist in the four groups of rats, due to endothelial NO release (entirely in females and in part in males); (3) androgen also seems to modulate the response by stimulating the release of an endothelial contracting factor, probably a prostanoid; and (4) the endothelium of males has a greater capacity than that of comparable females for negative regulation of the tension generated by the underlying vascular smooth muscle. PMID- 10369791 TI - Activation of haemostasis by exercise, mental stress and adrenaline: effects on platelet sensitivity to thrombin and thrombin generation. AB - Stress-induced activation of haemostasis may be involved in the triggering of acute coronary syndromes. We compared the effects of mental stress, dynamic exercise and adrenaline infusion on platelet sensitivity to thrombin using flow cytometric analysis of platelet fibrinogen binding in whole blood, and platelet aggregability using filtragometry ex vivo, in healthy volunteers. Furthermore, we assessed thrombin generation [prothrombin fragment 1+2 (F1+2) and thrombin antithrombin complexes in plasma] and thrombin activity (fibrinopeptide A in plasma). Exercise (bicycle ergometry) enhanced thrombin-induced platelet fibrinogen binding (P<0.05) and platelet aggregability (P<0.01), and elevated F1+2, thrombin-antithrombin complexes and fibrinopeptide A (P<0.05 for all three). Adrenaline infusion enhanced thrombin-induced platelet fibrinogen binding and platelet aggregability (P<0.05), and elevated thrombin-antithrombin complexes (P<0.05), whereas F1+2 and fibrinopeptide A levels were not significantly affected. Mental stress increased platelet sensitivity to high concentrations of thrombin only, and produced small increases in levels of thrombin-antithrombin complexes. Time control experiments showed no important changes with repeated measurements during rest. Platelet responses to exercise and adrenaline were reversible, with recovery 60 min later. Thus, heavy exercise and high levels of adrenaline reversibly increased platelet aggregability and platelet sensitivity to thrombin, and enhanced thrombin formation; the effects were most pronounced during exercise. Mental stress only weakly affected these parameters. PMID- 10369792 TI - Endothelial haemostatic factors are associated with progression of urinary albumin excretion in clinically healthy subjects: a 4-year prospective study. AB - A slightly elevated urinary albumin excretion rate (UAER), above 5-10 microgram/min, is a predictor of atherosclerotic cardiovascular disease. Endothelial dysfunction is an important early feature of atherosclerosis. The plasma concentration of von Willebrand factor (vWF), a potential marker of endothelial dysfunction, predicts a subsequent increase of UAER in patients with diabetes. The aim of this study is to test the hypothesis that high concentrations of vWF as well as other haemostatic factors predict progression of UAER in clinically healthy subjects. UAER was measured together with selected markers of haemostatic function-vWF, tissue plasminogen activator (tPA), plasminogen activator inhibitor, factor VII and fibrinogen-in healthy volunteers aged 40-65 years. After a mean follow-up of 4.1 years, 64 of 74 agreed to a re examination including re-measurement of UAER. Baseline vWF and tPA were both positively correlated to the change in UAER during follow-up (r=0.26, P=0.04 and r=0.40, P=0.001 respectively). The mean UAER increased significantly by 7.6 microgram/min and 7.5 microgram/min respectively in subjects with vWF and tPA above the medians at baseline (P=0.01 and P=0.003 respectively), whereas no changes in UAER were seen in subjects with vWF and tPA below the medians. Subjects with high tPA were also characterized by an excess of other cardiovascular risk factors at baseline. No significant differences in these risk factors were present between subjects with high or low vWF. High plasma concentrations of vWF and tPA are associated with progression of UAER in clinically healthy subjects. Both vWF and tPA are secreted by endothelial cells and the results suggest that endothelial dysfunction leads to progression of UAER. PMID- 10369793 TI - Contractile properties of rat skeletal muscles following storage at 4 degrees C. AB - The purpose of this study was to assess the potential of preservation solutions for protecting skeletal muscle function during storage at 4 degrees C. The soleus and the cutaneus trunci (CT) from the rat were stored for 2, 8 or 16 h at 4 degrees C in University of Wisconsin solution (UW), HTK-Bretschneider solution (HTK) or Krebs-Henseleit solution (KH). After storage, muscles were stimulated electrically to analyse the isometric contractile properties, such as the maximum tetanic tension (P(0)). Histological analysis was also performed. In separate experiments, the effect of the diffusion distance on muscle preservation was studied by bisecting the soleus. After 8 h of storage in UW or HTK, the contractile properties of the CT were similar to those of the control, whereas those of the soleus were reduced (P(0) values of 16% and 69% of control in UW and HTK respectively). At 16 h, the contractile properties of the CT (P(O) 28%) were again better preserved than those of the soleus (P(0) 9%). Muscle function deteriorated most after storage in KH (P(0) at 16 h: soleus, 3%; CT, 17%). The bisected soleus was equally well preserved as the CT (P(O) of bisected soleus at 8 h in UW and HTK: 86%). The functional data corresponded well with the histological data, which showed increasing muscle fibre derangement with increasing storage time. For both muscles and all solutions, the threshold stimulus current increased with increasing storage time (control, 0.1 mA; 16 h, 1.2 mA) and was strongly correlated with the deterioration in contractile properties. It is concluded that, at 4 degrees C, muscle is preserved better in UW and HTK (intracellular-like solutions) than in KH (extracellular-like solution). The soleus and CT were best protected in HTK. The diffusion distance is a critical factor for successful preservation of muscle function at 4 degrees C. The reduced function after 16 h of storage at 4 degrees C was caused by hypercontraction and necrosis of about 25% of the muscle fibres, and by deterioration of the electrical component of excitation-contraction coupling of the remaining fibres. PMID- 10369794 TI - Phagocyte functions in stressed rats: comparison of modulation by glutamine, arginine and ornithine 2-oxoglutarate. AB - The effects of diets supplemented with 6.8 mmol.day-1.kg-1 glutamine, arginine or ornithine 2-oxoglutarate [ornithine alpha-ketoglutarate (OKG), a precursor of both glutamine and arginine] on phagocyte functions [i.e. H2O2 production by leucocytes and secretion of tumour necrosis factor alpha (TNFalpha) by stimulated macrophages] of stressed rats were studied. The relationship between the immunological effects of these amino acids and their plasma and tissue (muscle and intestine) concentrations was also explored. The catabolic model used consisted of injections of dexamethasone (DEX; 1.5 mg.day-1.kg-1) for 5 days. As previously described, DEX suppressed TNFalpha secretion in stimulated macrophages. Supplementation with arginine or OKG, but not glutamine, was able to counteract the DEX effect on TNFalpha secretion. Glutamine, arginine and OKG supplementation increased H2O2 production by monocytes and polymorphonuclear neutrophils from DEX-treated rats. All DEX-treated rats showed plasma and muscle glutamine depletion and also a decrease in the concentration of arginine in the gastrocnemius. Supplementation with glutamine, arginine or OKG was not able to counteract these depletions. It was concluded that glutamine, arginine and OKG improve phagocyte responses during stress, and that glutamine depletion is not necessarily associated with dysimmunity, since no correlation between glutamine tissue pools and the immune state was observed. PMID- 10369795 TI - Effect of externally applied focused acoustic energy on clot disruption in vitro. AB - Application of low-frequency ultrasound for clot disruption has been suggested as a potential therapy to enhance thrombus dissolution, but the optimal mode for delivery of ultrasound with clot-disruptive properties has not yet been extensively explored. Target-specific effects are desirable and may be accomplished by focusing the ultrasound. Adequate focusing, however, requires a short wavelength. The aim of this study was to compare the clot-disruptive effects of different modalities of focused acoustic energy. An in vitro model (10 blood clots for each modality) was used to test the clot-disruptive capacity of (i) shock waves generated in an electrohydraulic lithotriptor; (ii) focused continuous ultrasound of frequency 1.1 MHz, delivered from a specially constructed piezoelectric transducer; and (iii) focused pulse-modulated ultrasound of frequency 1.1 MHz delivered from the same transducer. Exposure to 30 s of focused pulse-modulated ultrasound caused a marked reduction (99+/-2%) in clot weight compared with 30 shock waves (11+/-5%) or 30 s exposure to focused continuous wave ultrasound (11+/-6%) (P<0.0001). The observed marked and rapid disruptive effect on blood clots of focused high-frequency ultrasound indicates an alternative approach for external ultrasound-mediated thrombus destruction in vivo. The focused pulse-modulated technique has potential to exhibit the desired effect in a well-defined target volume and provides the means for control of the average power. PMID- 10369796 TI - Hypophosphatasia: diagnostic application of linked DNA markers in the dominantly inherited adult form. AB - Hypophosphatasia is a rare disease characterized by low serum levels of tissue non-specific alkaline phosphatase (TNSALP) and a spectrum of skeletal disease varying from the severest form with death in utero to mild with no clinical abnormality in adults. Currently, the diagnosis of hypophosphatasia is made on the basis of clinical findings, radiography, low serum alkaline phosphatase levels and raised abnormal phosphorylated metabolites; there are elevations in serum pyridoxal 5'-phosphate, urinary phosphoethanolamine and inorganic pyrophosphate. In borderline cases the biochemical diagnosis remains uncertain. Prenatally, diagnosis is made using radiography and ultrasonography together with chorionic villus tissue biopsy, in which TNSALP levels are measured using an antibody-based assay. Since hypophosphatasia results from mutations in the TNSALP gene we have, for the first time in two U.K. families, undertaken restriction fragment length polymorphism (RFLP) analysis using three intragenic RFLPs for BclI and MspI at the ALPL locus. One family was informative, and a mutant-allele specific haplotype with respect to three RFLPs was defined. In the other family the disease was shown to segregate with one allele of the BclI RFLP, but the MspI RFLPs were not informative. The disease segregated in the two families with different alleles of the BclI RFLP, suggesting that the mutations are likely to be different. We confirm that DNA analysis is likely to be the way ahead for diagnosing hypophosphatasia, and that standardized screening methods need to be developed for detecting mutations in these and other families. PMID- 10369797 TI - Changes in muscle strength in women following the menopause: a longitudinal assessment of the efficacy of hormone replacement therapy. AB - The effects of hormone deficiency at the menopause on muscle strength was examined in 10 healthy middle-aged women (1-3 years post-menopause) in a longitudinal trial over 39 weeks. Performance was compared with that of age matched females (n=11) taking a course of hormone replacement therapy (HRT). Muscle strength of the quadriceps was measured isometrically at 90 degrees of knee flexion and at angular velocities of 1.05, 2.09 and 3.13 rad/s using an isokinetic dynamometer. Hand grip strength was assessed by means of a portable dynamometer. Measurements were taken every 13 weeks for 39 weeks. Significant decreases in isometric strength (-10%) and dynamic leg strength at 1.05 rad/s ( 9%) were found in the post-menopausal women over 39 weeks. There was no change in strength in the HRT group. There were also no changes in leg strength at higher angular velocities or in grip strength for either the post-menopausal group or those taking HRT. While HRT preserved muscle strength, there was no evidence of a strengthening effect on skeletal muscle within this short period of treatment. A rapid loss of leg strength occurs post-menopausally in hormone-depleted women. HRT may offer protection against muscle weakness, although the hormone responsible for regulating strength is not evident using this model. PMID- 10369799 TI - The renal endothelin system in the Prague hypertensive rat, a new model of spontaneous hypertension. AB - In a new model of spontaneous hypertension, namely the Prague hypertensive rat (PHR), hypertension is transferred with a kidney transplanted from the PHR to its normotensive counterpart (PNR) by an as yet unknown mechanism. One candidate may be endothelin (ET), since this potent vasoconstrictor affects vascular tone, renal haemodynamics and renal excretory function, and all members of this peptide family are located within the kidney and act in an autocrine/paracrine fashion. In the present study we investigated, in the renal tissue of PHRs and PNRs: (1) preproET-1 and preproET-3 mRNAs as well as ET-1 and ET-3 peptide distribution, (2) endothelin-converting enzyme (ECE)-1 mRNA expression, and (3) ET receptors and their characteristics in membranes of glomeruli and papillae. In addition, plasma ET concentration and urinary ET excretion were determined. Quantitative measurements by competitive reverse transcription-polymerase chain reaction revealed ET-1 mRNA levels in the renal cortex from PHRs and PNRs of 1.09+/-0.13 and 1. 29+/-0.18 amol/microgram of total RNA respectively, and in red medulla of 2.72+/-0.82 and 3.30+/-0.68 amol/microgram respectively. In contrast, renal papilla from PHRs showed significantly lower levels of preproET-1 mRNA (1.81+/ 0.64 amol/microgram of total RNA, compared with 4.25+/-0.82 amol/microgram in PNRs; each n=5; P<0.05). The ET-1 peptide concentration in papillary tissue was also significantly lower in PHRs than in PNRs (120.2+/-30.8 and 491.3+/-53.4 fmol/mg of protein respectively; n=5; P<0.01), whereas it was similar in cortex and medulla from PHRs and PNRs. The preproET-3 mRNA content in renal tissue was much lower than that of preproET-1 mRNA. It was significantly higher in red medulla from PHRs compared with that from PNRs (0.25+/-0.05 and 0.13+/-0.02 amol/microgram of total RNA respectively; P<0.05), but was similar in papillae of PHRs and PNRs (0.04+/-0.02 and 0.05+/-0.01 amol/microgram respectively; n=5). Cortical preproET-3 mRNA was at the lower limit of detection. Similarly, the ET-3 peptide concentration was slightly but significantly higher in the red medulla of PHRs compared with PNRs (15.4+/-2.0 and 8.8+/-0.8 fmol/mg of protein respectively; n=5; P<0. 05), whereas no differences in ET-3 peptide concentration were found in papillae from PHRs and PNRs. ECE-1 mRNA levels were similar in the renal cortex, red medulla and papillae from PHRs and PNRs, ranging between 0.34+/ 0.03 and 0.56+/-0.12 amol/microgram of total RNA. Of the total ET receptors in glomerular membranes, 39% were ETA receptors, whereas papillary membranes contained exclusively ETB receptors. PHRs and PNRs showed similar Bmax and Kd values for ET-1 in renal glomerular membranes (Bmax, 6.5+/-1.3 and 4.9+/-1.2 pmol/mg of protein respectively; Kd, 0.69+/-0.10 and 0.56+/-0.10 nM respectively) and papillary membranes (Bmax, 9.7+/-1.1 and 11.3+/-1. 6 pmol/mg of protein respectively; Kd, 0.30+/-0.04 and 0.42+/-0.07 nM respectively). Plasma ET-1/2 concentrations (10.4+/-1.3 and 12. 2+/-1.2 fmol/ml in PHRs and PNRs respectively) and urinary ET-1 excretion (3.1+/-0.3 and 3.0+/-0.2 pmol/24 h in PHRs and PNRs respectively) were similar in hypertensive and normotensive rats. In summary, although tissue levels of preproET-3 mRNA were very low in the kidney, significantly greater amounts of preproET-3 mRNA and ET-3 peptide were found in medullary tissue from PHRs compared with PNRs, a finding that awaits further investigation. In contrast, the preproET-1 mRNA content and ET-1 peptide concentration were significantly lower in papillary tissue from PHRs compared with PNRs. Decreased synthesis of ET-1, which normally antagonizes the action of [Arg8]vasopressin, may allow increased water (and sodium) reabsorption at the level of the inner medullary collecting duct. This intrinsic defect of the kidney in the PHR may contribute to hypertension in this model, and may transmit high blood pressure on transplantation of the 'hypertensive' kidney i PMID- 10369800 TI - Effect of nebulized epoprostenol (prostacyclin) on exhaled nitric oxide in patients with pulmonary hypertension due to congenital heart disease and in normal controls. AB - Inhaled epoprostenol (prostacyclin) may be used in the treatment of severe pulmonary hypertension, improving oxygenation and reducing pulmonary artery pressures. We have observed symptomatic benefits of epoprostenol in patients with congenital heart disease that extend beyond acute haemodynamic effects of the drug, which has a short biological half-life. The aim of this study was to examine the effects of epoprostenol in patients and normal subjects on exhaled nitric oxide (eNO), based on the hypothesis that the drug may alter the resting vasoconstrictor/vasodilator balance. Nine patients with pulmonary hypertension complicating left-to-right cardiac shunts and nine healthy controls received 100 microgram of nebulized epoprostenol. Exhaled eNO was measured, using a chemiluminescence method, before, immediately after and 18 h after nebulization. There was no significant difference between the two groups in baseline eNO or eNO immediately following nebulized epoprostenol. Epoprostenol produced a delayed elevation in eNO 18 h after nebulization in patients, but not in normal controls. This study supports the concept that epoprostenol, while having no effect on the normal pulmonary circulation, acts on the hypertensive circulation via a mechanism that may result in a delayed alteration of vasoconstrictor/vasodilator balance. PMID- 10369798 TI - Effects of treatment of rheumatoid arthritis patients with an antibody against tumour necrosis factor alpha on reticuloendothelial and intrapulmonary granulocyte traffic. AB - The therapeutic effects of a monoclonal antibody against tumour necrosis factor alpha (TNFalpha) were evaluated objectively in 10 patients with rheumatoid arthritis by 111In-labelled granulocyte imaging before and after treatment, and compared with changes in granulocyte kinetics with respect to the liver, spleen and lungs. Anti-TNFalpha resulted in a decrease in the size of the whole-body pool of marginating granulocytes, as reflected by a significant increase in the 30 min intravascular recovery of labelled granulocytes from 40% (S.D. 10) to 47% (S.D. 16) of injected activity (P<0.02). The 111In contents of the spleen, liver and lungs were unchanged, so the origin of the increment in recovery was presumed to be a reduction in granulocyte margination in inflamed synovium, although this was not quantifiable. The sizes of the granulocyte pools in the liver and lungs, expressed as the 111In content of the organ per unit of circulating 111In labelled cells, were not significantly different after treatment, but the splenic granulocyte pool decreased by 16% (S.D. 19) (P<0.05). Individual changes in the size of the splenic pool showed no significant correlation with corresponding changes in 30 min recovery or with corresponding indices of inflammation (24 h 111In-granulocyte joint activity and C-reactive protein). We conclude that anti TNFalpha produces an obvious resolution in inflammatory joint activity that is accompanied by an increased circulating component of the total blood granulocyte pool, as a result of decreased margination at sites of inflammation. Anti TNFalpha may also produce a specific decrease in splenic granulocyte pooling, independent of any anti-inflammatory effects, although a similar decrease in the lungs, which might be anticipated as a result of reduced cytokine-induced granulocyte activation, could not be detected. PMID- 10369801 TI - Altered cardiovascular haemodynamics and baroreceptor-heart rate reflex in adult sheep after prenatal exposure to dexamethasone. AB - Numerous epidemiological studies, together with mounting evidence from studies in animals, point to a correlation between an adverse intrauterine environment and the early onset of cardiovascular and metabolic diseases later in life. We were the first to show that sheep exposed to dexamethasone (0.28 mg.kg-1.day-1 for only 2 days) at the end of the first month of pregnancy (PTG1), but not those exposed at the end of the second month of pregnancy (PTG2), had a higher basal mean arterial pressure (MAP) 19 months after birth. In the present study we report the MAP, cardiovascular haemodynamics and baroreflex sensitivity in these animals at 40 months of age. MAP in the PTG1 group was significantly higher than in the control group (91+/-1 mmHg and 81+/-1 mmHg respectively; P<0.001) and also when compared with the PTG2 group (82+/-1 mmHg; P<0.001). There was a significant increase in cardiac output in the PTG1 group compared with the control group (108+/-2 and 96+/-4 ml.min-1.kg-1 respectively; P<0.05). The increase in cardiac output in the PTG1 group was due to an increase in stroke volume (1.82+/-0.08 ml.kg-1. beat-1, compared with 1.46+/-0.06 ml.kg-1.beat-1 in the control group; P<0.05), but not in heart rate. In the hypertensive group of animals (PTG1), there was a rightward shift of the baroreflex curve. In group PTG2 (the normotensive group of animals), a lower gain was found before and during propranolol treatment. The decrease in gain of the baroreflex was not associated with changes in heart rate range, suggesting an impairment in the central processing of the baroreceptor signals. Thus sheep fetuses exposed to dexamethasone for only 2 days at the end of the first month of gestation have high blood pressure (dependent upon the increase in cardiac output) and a reset of the baroreflex at 40 months of age. Animals that have received prenatal dexamethasone closer to mid-gestation, although normotensive with normal cardiac output, showed an altered baroreceptor-heart rate response. PMID- 10369802 TI - Elevated activity of semicarbazide-sensitive amine oxidase in blood from patients with skeletal metastases of prostate cancer. AB - The semicarbazide-sensitive amine oxidases constitute a group of copper containing enzymes whose physiological function is unclear. The enzymes are present in various tissues, including blood plasma. At present, the source of the plasma enzyme in humans is not known. Results of a recent study suggested that semicarbazide-sensitive amine oxidase is expressed in the skeleton, e.g. in the spine. Using an indirect autoradiographic method in mice, we provide evidence that semicarbazide-sensitive amine oxidase is present in high abundance in bone tissue. Specific activities of semicarbazide-sensitive amine oxidase were estimated in blood samples from subjects with femoral bone fractures. Moreover, enzyme activities were also measured in patients suffering from prostate cancer with skeletal metastases. The level of specific semicarbazide-sensitive amine oxidase activity in serum was significantly elevated in patients with skeletal metastases compared with both healthy controls and patients having prostate cancer without signs of skeletal metastases. Based on the results of the present study, we propose that semicarbazide-sensitive amine oxidase in blood plasma may originate, at least in part, from the skeleton. PMID- 10369803 TI - Blood coagulation, fibrinolysis and exercise. PMID- 10369804 TI - Endothelial cell damage and the development or progression of atherosclerosis. PMID- 10369805 TI - Human placental nitric oxide synthase activity is not altered in diabetes. AB - Endothelial nitric oxide synthase (NOS) protein and mRNA have been identified and calcium-dependent NOS activity has been measured in human placentae during normal pregnancy. Recently, mRNA and protein for the inducible isoform of NOS have been detected in placentae of women with gestational diabetes. The aim of this study was to determine whether calcium-independent (ciNOS) and/or total (tNOS) NOS activities were increased in placentae obtained after vaginal delivery or Caesarean section from women assigned to the following groups according to standard obstetric criteria: gestational diabetes, diabetes before pregnancy and non-diabetic controls. tNOS and ciNOS were assessed by measuring the conversion of [3H]L-arginine to [3H]L-citrulline in the three groups. Michaelis-Menten constants (Km) and maximum velocities of reaction (Vmax) were calculated using Lineweaver-Burk analysis for tNOS. There were no significant differences in either ciNOS, Vmax or Km values between any of the three groups (normal, ciNOS 12.7+/-1.6%, Vmax 16.6+/-3.3 pmol.min-1.mg-1 protein, Km 15.30+/-2.6 micromol/l; gestational diabetes, ciNOS 15.4+/-1.4%, Vmax 14.8+/-5.2 pmol.min-1. mg-1 protein, Km 10.5+/-1.7 micromol/l; diabetes before pregnancy, ciNOS 13.4+/-1.1%, Vmax 14.9+/-3.4 pmol.min-1.mg-1 protein, Km 17. 7+/-2.2 micromol/l). The presence of macrosomia did not affect tNOS activity in those with diabetes before pregnancy, and glycosylated haemoglobin levels measured between weeks 27 and 39 were not correlated with ciNOS activity. The results from the present study do not provide evidence for increased placental tNOS or ciNOS activities in pregnancies complicated by gestational diabetes or diabetes present before pregnancy. PMID- 10369806 TI - Hydroxyethyl radicals in ethanol hepatotoxicity. AB - Alcoholic patients and experimental animals exposed to ethanol display biochemical signs of oxidative damage, suggesting a possible role of free radicals in causing some of the toxic effects of alcohol. The use of electron spin resonance (ESR) spectroscopy associated with spin trapping technique has demonstrated that hydroxyethyl radicals are generated during ethanol metabolism by the microsomal monoxygenase system, involving the alcohol-inducible cytochrome P450 2E1 (CYP2E1). Recent observations in rats fed intragastrically with a high fat diet containing ethanol indicate that the formation of hydroxyethyl radicals is associated with stimulation of lipid peroxidation and development of liver damage. Moreover, by alkylating liver proteins, and particularly CYP2E1, hydroxyethyl free radicals are also capable of inducing the production of specific antibodies which can be observed in ethanol-fed animals as well as in patients abusing alcohol. Recent studies have demonstrated that hydroxyethyl radical-derived antigens are exposed on the plasma membranes of hepatocytes exposed to ethanol where are able to target antibody-dependent cell-mediated immunotoxic reactions towards liver cells. Thus, beside to contribute to alcohol mediated oxidative damage, hydroxyethyl free radicals can contribute by immunological mechanisms to cause hepatocellular lesions associated with alcohol abuse. PMID- 10369807 TI - Ethanol, oxidative stress, reactive aldehydes, and the fetus. AB - The fetotoxic effects of maternal ethanol (E) consumption have been documented for over two decades, yet the mechanisms underlying this devastating phenomenon remain uncertain. The wide variety of cellular/biochemical effects of E on fetal tissues is itself a puzzle and strongly suggests that fetotoxic responses to E reflect a multifactorial setting. Many of these responses can be conceptually connected to effects on membrane structure and function. Representative of this, are studies in our laboratory documenting E effects on fetal cell replication, membrane transport systems, membrane fluidity, Na+-K+ pump expression, and EGF receptor expression. Recent studies have provided evidence that oxidative stress may be one mechanism by which E produces these membrane-related events. We initially observed E-induced oxidative stress in cultured fetal rat hepatocytes, the latter exhibiting morphological and biochemical signs of mitochondrial damage. E increased H2O2, O2-, lipid peroxidation products, along with signs of membrane damage. Supplementation with antioxidants or agents that enhance glutathione stores reversed these effects. E was found to inhibit activities of mitochondrial respiratory chain components (a potential source of the enhanced levels of H2O2, and O2-) and this could be reversed by antioxidant treatment. Subsequent studies have documented oxidative stress and membrane lipid peroxidation in fetal brain and liver (gestation day 19) following a two day maternal E consumption and in gestation day 14 and 17 "embryos" immediately following a single dose of E to the pregnant dam. The means by which E can induce oxidative stress in fetal cells is under investigation. We have examined effects of E on activities of key antioxidant enzymes and found no depressant responses. However, the low levels of antioxidants in fetal tissues and an exaggerated response of fetal mitochondria to prooxidant stimulation in vitro, suggest that fetal cells are strongly predisposed to oxidative stress. Additionally, recent studies have suggested that fetal tissues are likewise prone to the formation and subsequent accumulation of at least one toxic lipid peroxidation product, 4 hydroxynonenal. We conclude that maternal E consumption induces oxidative stress in fetal tissues and that this is responsible for some toxic responses to E. Additionally, the low antioxidant defenses in fetal tissues and accumulation of toxic aldehyde products of lipid peroxidation predispose the fetus to oxidative damage. PMID- 10369808 TI - The retinoblastoma protein-interacting zinc finger gene RIZ in 1p36-linked cancers. AB - Mutations or changes in protooncogenes and tumor suppressor genes are casually linked to human cancer pathogenesis. The RIZ gene is isolated based on the capacity its gene products to bind to the retinoblastoma tumor suppressor protein. Consistent with a potential role in the Rb pathway, RIZ may play an important role in human cancer pathogenesis. RIZ maps to human chromosome band 1p36, a region commonly altered in many types of human cancers. RIZ is the founding member of the PR-domain family of zinc finger genes. Similar to other members of this family, RIZ is involved in human cancers in an unusual yin-yang fashion. Two products are produced from the RIZ locus which differ by the presence or absence of the PR domain; the PR-plus product RIZ1 is commonly lost or underexpressed whereas the PR-minus product RIZ2 is always present in cancer cells. This yin-yang imbalance in the amount of the two RIZ products may be an important cause of malignancy. PMID- 10369810 TI - Nitric oxide in acute ischaemic stroke: a target for neuroprotection. PMID- 10369809 TI - NF-kappa B, liposomes and pathogenesis of hepatic injury and fibrosis. AB - The liver injury caused by hepatotoxins is characterized by varying degrees of hepatocyte degeneration and cell death via either apoptosis or necrosis. Generation of reactive intermediate metabolites from the metabolism of toxins and the occurrence of reactive oxygen species (ROS) during the inflammatory reaction account for a variety of pathophysiologic pathways which lead to cell death. This process can then evoke acute or chronic inflammatory responses if the injury is sustained, and these pathologic alterations eventually progress to cirrhosis. Understanding the function of transcription factors, such as nuclear factor kappa?B (NF-kappa B), in acute liver injury may provide some answers to the molecular mechanisms of toxic insults. Liposomes have been used as vehicles for drug delivery and gene therapy and they have been shown to have substantial potential in the targeting of specific cell types of the liver. Thus, the use of liposomes may improve targeting efficacy in the treatment of a variety of liver diseases. PMID- 10369811 TI - Carcinoma associated paraneoplastic peripheral neuropathy. PMID- 10369812 TI - Right hemisphere contributions to attention and intention. PMID- 10369813 TI - New hope for patients with pure lower motor neuron syndromes. PMID- 10369815 TI - Armauer Gerhard Heinrik Hansen (1841-1912). PMID- 10369814 TI - Carcinoma associated paraneoplastic peripheral neuropathies in patients with and without anti-onconeural antibodies. AB - OBJECTIVE: When to suspect a paraneoplastic disorder is a puzzling problem that has not recently been studied in a large series of patients referred for peripheral neuropathy. METHODS: From 422 consecutive patients with peripheral neuropathy, 26 were analysed who concomitantly had carcinoma but no tumorous infiltration, drug toxicity, or cachexia. Their clinical, pathological, and electrophysiological data were analysed according to the presence of anti onconeural antibodies, the latency between presentation and cancer diagnosis, and the incidence of carcinoma in the corresponding types of neuropathy of the population of 422 patients. RESULTS: Seven patients (group I) had anti-onconeural antibodies (six anti-Hu, one anti-CV2) and 19 did not (groups IIA and B). In group I, subacute sensory neuropathy (SSN) was the most frequent but other neuropathies including demyelinating neuropathies were present. Patients in group II A had a short latency (mean 7.88 months), and a rapidly and usually severe neuropathy which corresponded in 11/14 to an established inflammatory disorder including neuropathy with encephalomyelitis, mononeuritis multiplex, and acute or chronic inflammatory demyelinating polyneuropathy (CIDP). Patients in group IIB had a long latency (mean 8.4 years) and a very chronic disorder corresponding in four of five to an axonal non-inflammatory polyneuropathy. In this population, the incidence of carcinoma occurring with a short latency was 47% in sensory neuronopathy, 1.7% in Guillain-Barre syndrome, 10% in mononeuritis multiplex and CIDP, and 4.5% in axonal polyneuropathy. CONCLUSIONS: Paraneoplastic neuropathies associated with carcinoma are heterogeneous disorders. Neuropathies occurring with a long latency with tumours probably resulted from a coincidental association. Neuropathies which occurred within a few years of the tumour evolved rapidly and corresponded mostly to inflammatory disorders. As dysimmune neuropathies are probably paraneoplastic in a limited number of cases, patients with these disorders should probably not be investigated systematically for carcinoma in the absence of anti-onconeural antibodies, except when the neuropathy is associated with encephalomyelitis and probably with vasculitis. Questions remain concerning CIDP. PMID- 10369816 TI - Use of human intravenous immunoglobulin in lower motor neuron syndromes. AB - OBJECTIVE: To determine whether patients with the clinical phenotype of multifocal motor neuropathy but without the electrophysiological criteria for conduction block would respond to intravenous immunoglobulin (IVIg). METHODS: Ten patients were selected with a slowly progressive, asymmetric, lower motor neuron disorder, and were treated prospectively with IVIg at a dose of 2g/kg over 5 days. All subjects had neurophysiological testing to look for evidence of conduction block before treatment. Muscle strength was assessed by MRC grades and hand held myometry, measuring pinch and grip strength. A 20% increase in both pinch and grip myometry was considered a positive response. RESULTS: In no patient was conduction block detected. Four of the 10 patients showed a positive response to IVIg, with the best response occurring in two patients who presented with weakness but without severe muscle wasting. Three of the four responders have continued to receive IVIg for a mean period of 17 months (range 15-24 months), with continued effect. The response to IVIg was not related to the presence of anti-GM1 antiganglioside antibodies, but responders had a selective pattern of muscle weakness and normal (>90% predicted) vital capacity. CONCLUSION: The findings suggest that a course of IVIg should be considered in patients with the clinical phenotype of multifocal motor neuropathy but without neurophysiological evidence of conduction block. PMID- 10369817 TI - Evidence for cortical dysfunction in clinically non-demented patients with Parkinson's disease: a proton MR spectroscopy study. AB - OBJECTIVES: To investigate whether proton magnetic resonance spectroscopy (1H MRS) can detect cortical dysfunction in non-demented patients with Parkinson's disease, and to correlate changes with cognitive function on formal neuropsychological testing. METHODS: Multivoxel 1H MRS was performed in 17 patients with levodopa treated idiopathic Parkinson's disease with out clinical dementia, and 10 age match ed control subjects. Measurements of N-acetylaspartate (NAA)/choline (Cho), NAA/creatine+phosphocreatine (Cr), and Cho/Cr were obtained from right and left temporoparietal cortex and occipital cortex. Fourteen patients with Parkinson's disease underwent a full battery of neuropsychological testing including performance and verbal subtests of the WAIS-R, Boston naming test, FAS test, and California verbal learning test. RESULTS: There were significant temporoparietal cortex reductions in NAA/Cr ratios in right and left averaged spectra of the patients with Parkinson's disease (p=0.012 after Bonferroni correction) and in spectra contralateral to the worst clinically affected limbs of the patients with Parkinson's disease compared with controls (p = 0.003 after Bonferroni correction). There was a significant correlation between reduction in NAA/Cr ratios and measures of global cognitive decline, occurring independently of motor impairment (p=0.019). CONCLUSIONS: This study suggests that 1H MRS can detect temporoparietal cortical dysfunction in non-demented patients with Parkinson's disease. Further longitudinal studies are needed to investigate whether these 1H MRS changes are predictive of future cognitive impairment in the subset of patients with Parkinson's disease who go on to develop dementia, or occur as part of the normal Parkinson's disease process. PMID- 10369818 TI - Learning in Parkinson's disease: eyeblink conditioning, declarative learning, and procedural learning. AB - OBJECTIVE: To assess the degree of learning ability in Parkinson's disease. METHODS: Three different learning tasks: eyeblink classical conditioning with delay and trace paradigms, the California verbal learning test (CVLT), and a serial reaction time task (SRTT) were studied in patients with Parkinson's disease and normal (control) subjects. RESULTS: In the eyeblink conditioning tasks, both patients and normal subjects showed significant learning effects without between group differences. In the CVLT, patients remembered significantly fewer words than normal subjects in both short term and long term cued recall tasks. In the SRTT, normal subjects had significantly reduced response time and error rates across blocks of repeated sequence trials, whereas patients had significantly reduced error, but not response time rates. CONCLUSION: Impairment of nigrostriatal pathways selectively affects performance in complex learning tasks that are competitive and require alertness such as the SRTT, but not in simple learning procedures such as eyeblink conditioning. PMID- 10369819 TI - Ipsilesional neglect: behavioural and anatomical features. AB - OBJECTIVE: To learn more about the behavioural and anatomical features of ipsilesional neglect. METHODS: Thirty consecutive patients with spatial neglect were tested on cancellation and line bisection tasks. To learn if patients with ipsilesional neglect demonstrate the sensory-attentional or motor-intentional type of neglect, a video apparatus was used that dissociates these determinants. RESULTS: Five patients showed evidence of ipsilesional neglect. This phenomenon was seen only on the line bisection task. All patients with ipsilesional neglect had lesions involving frontal-subcortical regions. Although ipsilesional neglect evolved from early in three of five cases, the other patients displayed ipsilesional neglect without initial contralateral neglect, suggesting that ipsilesional neglect cannot be fully attributed to a compensatory strategy. The results of the tests that used the video apparatus indicate that right sided frontal or subcortical injury may induce contralateral attentional or intentional "approach" behaviours. CONCLUSIONS: Ipsilesional neglect is most often associated with frontal-subcortical lesions, cannot be entirely attributed to a compensatory strategy, and may be induced by an attentional bias, an intentional bias, or both. PMID- 10369820 TI - Dysautonomia after traumatic brain injury: a forgotten syndrome? AB - OBJECTIVES: To better establish the clinical features, natural history, clinical management, and rehabilitation implications of dysautonomia after traumatic brain injury, and to highlight difficulties with previous nomenclature. METHODS: Retrospective file review on 35 patients with dysautonomia and 35 sex and Glasgow coma scale score matched controls. Groups were compared on injury details, CT findings, physiological indices, and evidence of infections over the first 28 days after injury, clinical progress, and rehabilitation outcome. RESULTS: the dysautonomia group were significantly worse than the control group on all variables studied except duration of stay in intensive care, the rate of clinically significant infections found, and changes in functional independence measure (FIM) scores. CONCLUSIONS: Dysautonomia is a distinct clinical syndrome, associated with severe diffuse axonal injury and preadmission hypoxia. It is associated with a poorer functional outcome; however, both the controls and patients with dysautonomia show a similar magnitude of improvement as measured by changes in FIM scores. It is argued that delayed recognition and treatment of dysautonomia results in a preventable increase in morbidity. PMID- 10369821 TI - Long term effects of refractory temporal lobe epilepsy on cognitive abilities: a cross sectional study. AB - OBJECTIVE: Intractable epilepsy is related to various transient and chronic brain electric and neurochemical disturbances. There is increasing evidence that chronic epilepsy induces secondary neuronal metabolic and structural decline. However, there is no convincing evidence that the cognitive abilities of patients deteriorate with increasing duration of intractable epilepsy. METHODS: To examine whether duration of refractory temporal lobe epilepsy (TLE) is related to generalised cognitive impairment, psychometric intelligence based on the full scale intelligence quotient (FSIQ, WAIS-R) was determined in 209 patients with unilateral TLE. For analyses of variance (ANOVA) patients were grouped into three categories: <15, 15-30, and >30 years of refractory TLE. RESULTS: An ANOVA and a multiple regression analysis showed that duration of TLE affects FSIQ. Patients with >30 years of TLE performed worse than patients with 15 or 30 years of TLE. The factors side of seizure origin and type of lesion on MRI did not reach significance. A second ANOVA including education as factor showed that in patients with higher educational attainment, the mean FSIQ was stable for a longer duration of TLE than in less educated patients. Retesting 6 months after anterior temporal lobectomy seizure free patients (n=85 of 127) had an higher FSIQ but showed a similar duration effect before and after anterior temporal lobectomy. The variables age at epilepsy onset, education, frequency of interictal epileptiform discharges, frequency of habitual and generalised seizures, serum concentration of antiepileptic drugs, and polypharmacy were statistically controlled. CONCLUSIONS: Psychometric intelligence of patients with a longer duration of refractory TLE were most severely impaired. Consequently, refractory TLE seems to be associated with slow but ongoing cognitive deterioration. It is assumed that epilepsy related noxious events and agents exhaust the compensatory capacity of brain functions. However, as in dementia and Alzheimer's disease, higher educational attainment as an indicator of higher brain reserve might delay the cognitive decline. PMID- 10369822 TI - Generation of scalp discharges in temporal lobe epilepsy as suggested by intraoperative electrocorticographic recordings. AB - OBJECTIVES: To study the variability, topography, polarity, duration, and incidence of interictal epileptiform discharges (EDs) in the scalp EEG and electrocorticogram (ECoG) from 16 patients with temporal lobe epilepsy who underwent surgical treatment. METHODS: Preoperative scalp EEGs during quinalbarbitone induced sleep were compared with preresection ECoGs obtained under general anaesthesia. The analysis was based on the initial ECoG record obtained before activation by intravenous thiopentone, and the EEG during stages I and II of sleep. RESULTS: On the scalp, 15 patients had a single discharge pattern, spikes were predominantly negative, EDs were of largest amplitude at the anterior temporal electrode in 13 patients and mean discharge incidence was 4.0 (SD 4.2) discharges/min. In ECoG recordings, nine patients had two independent ECoG patterns, the polarity of spikes was negative, positive-negative, or positive, the site of maximal amplitude varied greatly between subjects, discharge incidence was 7.3 (SD 3.9) discharges/min. There was no relation between the topography of the largest spikes on the scalp and in the ECoG. In 14 patients, scalp spikes showed statistically significant longer duration on the scalp than in the ECoG. In seven patients who had frequent widespread ECoG discharges, averaging spikes across ECoG channels generated spiky patterns of duration similar to that of scalp spikes. CONCLUSION: It seems that, in temporal lobe epilepsy, scalp discharges originate from widespread ECoG discharges and tend to produce a stereotyped pattern on the scalp with largest amplitudes at the anterior temporal electrodes. This is probably due to local anatomical peculiarities in the brain coverings, such as skull discontinuities, rather than to the location of neuronal generators within the temporal lobe. Due to spatiotemporal averaging, widespread cortical discharges which become asynchronous during propagation appear with increased duration and blunted waveform in the EEG, whereas sharply localised phenomena such as positive focal spikes are not recorded from the scalp. PMID- 10369823 TI - Predictors of mortality in patients with Alzheimer's disease living in nursing homes. AB - OBJECTIVES: To identify factors associated with mortality in patients with Alzheimer's disease, and to evaluate whether these factors vary according to severity of cognitive impairment. METHODS: Data were from the SAGE database which includes information on all residents admitted between 1992 and 1995 to all Medicare/ Medicaid certified nursing homes of five US states. We conducted a longitudinal follow up study (median 23 months) on 9264 patients aged 65 years and above with a diagnosis of Alzheimer's disease. Patient data including demographic characteristics, dementia severity, comorbidity, and other clinical and treatment variables were collected with the Minimum Data Set. Information on death was derived through linkage to Medicare files. Baseline characteristics were used to predict survival in univariate and multivariate Cox proportional hazard models. RESULTS: Overall mortality rate was 50%, with a first year rate of 25.7%. Increased age (risk ratio (RR) 1. 83; 95% confidence interval (95% CI) 1.65-2.03, for patients 85+ years), male sex (RR 1.81; 95% CI 1.70-1.94), limitation in physical function (RR 1.45; 95% CI 1.27-1.66), a condition of malnutrition (RR 1.31; 95%CI 1.23-1.39), the presence of pressure ulcers (RR 1.24; 95% CI 1.13-1.36), a diagnosis of diabetes mellitus (RR 1.32; 95% CI 1.21 1.43), and of cardiovascular diseases (RR 1.22; 95% CI 1. 14-1.30) were independent predictors of death, regardless of the severity of baseline dementia. Sensory problems (hearing and vision) and urinary incontinence were associated with increased mortality only among patients with less severe dementia. The presence of disruptive behaviour, aphasia, and a diagnosis of Parkinson's disease were not related to survival. African-Americans and other minority groups were less likely to die relative to white people. CONCLUSIONS: Age, sex, functional limitation, and malnutrition seem to be the strongest predictors of death for patients with Alzheimer's disease in nursing homes. Altogether, severity of dementia has no influence on survival, yet the predictive role of certain variables depends on the degree of impairment. Minority groups have a reduced risk of death relative to white people. PMID- 10369825 TI - Perforating branches from offending arteries in hemifacial spasm: anatomical correlation with vertebrobasilar configuration. AB - OBJECTIVE: In microvascular decompression for hemifacial spasm, the perforating branches around the facial nerve root exit zone occasionally complicate facial nerve decompression. In this context, the vertebrobasilar configuration was retrospectively correlated with the perforating branches. METHODS: Based on vertebral angiography, magnetic resonance angiography, and three dimensional computed tomographic angiography, 69 patients were divided into three groups, according to the anatomy of the vertebrobasilar system. In patients with the type I configuration, the vertebral artery on the affected side was dominant and had a sigmoidal course. The type II patients had the basilar artery curving mainly towards the affected side. The type III patients showed the basilar artery either running straight or curving toward the unaffected side. The relation of the anatomical configuration of these vessels with the perforating branches around the facial nerve exit zone was investigated. RESULTS: The posterior inferior cerebellar artery in type I patients (n=33) and the anterior inferior cerebellar artery in type II (n=5) and type III (n=31) patients were the most common offending arteries. More than half of the type I patients (n=20) showed no perforating branches around the facial nerve exit zone. However, the type II (n=3) and III patients (n=23) often showed one or more perforating branches around that region. CONCLUSIONS: The configuration of the vertebrobasilar system has a significant correlation with the presence of perforating branches near the site of microvascular decompression. These perforating vessels are often responsible for the difficulty encountered in mobilising the offending artery during the procedure. PMID- 10369824 TI - White matter lesions on magnetic resonance imaging in dementia with Lewy bodies, Alzheimer's disease, vascular dementia, and normal aging. AB - OBJECTIVES: Alzheimer's disease and vascular dementia are associated with an increase in changes in white matter on MRI. The aims were to investigate whether white matter changes also occur in dementia with Lewy bodies and to examine the relation between white matter lesions and the cognitive and non-cognitive features of dementia with Lewy bodies, Alzheimer's disease, and vascular dementia. METHODS: Proton density and T2 weighted images were obtained on a 1.0 Tesla MRI scanner in patients with dementia with Lewy bodies (consensus criteria; n=27, mean age=75.9 years), Alzheimer's disease (NINCDS/ADRDA; n=28, mean age=77.4 years), vascular dementia (NINDS/AIREN; n=25, mean age=76.8 years), and normal controls (n=26, mean age=76.2 years). Cognitive function, depressive symptoms, and psychotic features were assessed using a standardised protocol. Periventricular hyperintensities (PVHs), white matter hyperintensities (WMHs) and basal ganglia hyperintensities (BGHs) were visually rated blind to diagnosis using a semiquantitative scale. RESULTS: Periventricular hyperintensities were positively correlated with age and were more severe in all dementia groups than controls. Total deep hyperintensities scores (WMHs plus BGHs) were significantly higher in all dementia groups than controls and higher in patients with vascular dementia than those with dementia with Lewy bodies or Alzheimer's disease. In all patients with dementia, frontal WMHs were associated with higher depression scores and occipital WMHs were associated with an absence of visual hallucinations and delusions. CONCLUSION: In common with Alzheimer's disease and vascular dementia, PVHs and WMHs were significantly more extensive in dementia with Lewy bodies than in controls. This overlap between different dementias may reflect shared pathological mechanisms. The link between frontal WMHs and depression and the absence of occipital WMHs and psychotic symptoms has important implications for understanding the neurobiological basis of these symptoms. PMID- 10369826 TI - Autosomal dominant burning feet syndrome. AB - Familial burning feet syndrome inherited as an autosomal dominant trait has been described in only one family. Due to an associated sensory neuropathy the autosomal dominant burning feet syndrome was suggested to represent a variant form of hereditary sensory and autonomic neuropathy type I (HSAN I). Clinical, histopathological, and molecular genetic studies were performed in a large German kindred with autosomal dominant burning feet syndrome. The autosomal dominant burning feet syndrome was associated with a neuropathy predominantly affecting small unmyelinated nerve fibres. Linkage to the HSAN I locus on chromosome 9q22 and to the Charcot-Marie-Tooth disease type 2B (CMT 2B) locus on chromosome 3q13 q22 was excluded. The autosomal dominant burning feet syndrome is neither allelic to HSAN I nor to CMT 2B and thus represents a distinct genetic entity. PMID- 10369827 TI - Pupillographic findings in neglect. AB - OBJECTIVES: Unilateral sensory neglect has been attributed to various defects, including a hemispatial attention-arousal deficit. However, support for this hypothesis has only been indirect. Therefore, the purpose of this study was to further test the hemispatial attentional-arousal hypothesis by measuring pupillary response as an index of arousal. METHODS: There were two experimental subjects with neglect and six matched controls. Stimuli (Arabic numbers) were presented on the right, left, and centre of a screen. The subjects were asked to look at the number in the centre, on the right, or left of the screen while their pupil diameter was measured. RESULTS: Unlike the control subjects, the subjects with neglect, who were aware of the left sided stimuli, did not show a pupillary dilatation when they looked at the stimulus on the left. CONCLUSIONS: Although this study provides support for the hemispatial attention-arousal hypotheses of neglect, it does not preclude the possibility that other mechanisms may also be important. PMID- 10369828 TI - Clinical, neuropathological, and molecular study in two families with spinocerebellar ataxia type 6 (SCA6). AB - To clarify the clinical, neuropathological, and molecular characteristics of spinocerebellar ataxia type 6 (SCA6), two unrelated Japanese families with SCA6 were studied. A clinical feature of the two families was late onset "pure" cerebellar ataxia. Pathologically, three SCA6 brains consistently showed Purkinje cell dominant cortical cerebellar degeneration. Morphometric analysis showed that loss of the cerebellar granule cells and inferior olivary neurons were very mild compared with the severity of Purkinje cell loss. There was no obvious ubiquitin immunoreactive nuclear inclusions. All affected patients had identical expanded alleles, and the expansion was also homogeneously distributed throughout the brain without mosaicism. The present study showed that SCA6 is characterised by Purkinje cell dominant cortical cerebellar degeneration, highly stable transmission of the CAG repeat expansion, and lack of ubiquitin immunoreactive nuclear inclusions. PMID- 10369829 TI - Generalised muscular weakness after botulinum toxin injections for dystonia: a report of three cases. AB - Three patients are reported on who developed transient generalised weakness after receiving therapeutic doses of botulinum toxin for cervical dystonia (one case) and symptomatic hemidystonia (two cases) respectively. Clinical and electrophysiological findings were in keeping with mild botulism. All patients had received previous botulinum toxin injections without side effects and one patient continued injections without recurrence of generalised weakness. The cause is most likely presynaptic inhibition due to systemic spread of the toxin. Patients with symptomatic dystonia may be more likely to have this side effect and botulinum toxin injections in these patients should be carried out cautiously. PMID- 10369830 TI - Failure to confirm a synergistic effect between the K-variant of the butyrylcholinesterase gene and the epsilon4 allele of the apolipoprotein gene in Japanese patients with Alzheimer's disease. AB - To confirm a synergistic effect between the polymorphic K variant of the butyrylcholinesterase (BChE-K) gene and the epsilon4 allele of the apolipoprotein E (APOE) gene in Alzheimer's disease, the frequency of the BChE-K allele was re examined in a large series of Japanese patients with Alzheimer's disease and controls. Two hundred and three patients with Alzheimer's disease and 288 age and sex matched controls were genotyped by polymerase chain reaction and restriction fragment length polymorphism for BChE-K and APOE. No changes were found in the frequency of BChE-K, either in the Alzheimer's disease group as a whole (0.17 v 0.14; p=0.36) or in early (0.16 v 0.16; p=0.98) or late (0.17 v 0.13; p=0.24) onset patients compared with age matched controls. The study failed to confirm the findings of a previous study which found a significantly higher incidence of BChE-K in patients with Alzheimer's disease with APOE epsilon4 allele than in controls. In the Japanese population studied here, there was no association between BChE-K and Alzheimer's disease, nor an interaction between BChE-K and APOE epsilon4 allele. PMID- 10369831 TI - Spontaneous intralesional haemorrhage in dysembryoplastic neuroepithelial tumours: a series of five cases. AB - Five patients with dysembryoplastic neuroepithelial tumour (DNT) showing extensive secondary haemorrhage, a finding not previously associated with these neoplasms, are described. The clinical presentations, neuroimaging findings, and histopathological features of these patients are reviewed. One patient, a previously asymptomatic 12 year old girl, presented with an acute intracerebral haemorrhage into a DNT. A further four young adults with histories of intractable partial and generalised seizures dating from childhood showed significant chronic haemorrhages within DNT, the MRI appearances in one patient giving a false impression of a cavernoma. Histopathology disclosed vascular abnormalities within these tumours which, together with other factors discussed, may have predisposed these tumours to haemorrhage. PMID- 10369832 TI - Retrograde temporal order amnesia resulting from damage to the fornix. AB - Some amnesic patients show an impairment of temporal order memory that cannot be accounted for by content memory deficits. The performance of an amnesic patient on memory tasks assessing the patient's content and temporal memories for remotely acquired material is described, after a lesion including the bilateral anterior fornix and adjacent anterior thalamus. The patient displayed a deficit in the temporal order tasks for remotely acquired information. Neither frontal cognitive deficits nor recognition deficits can account for this patient's poor temporal memory. This retrograde temporal order memory impairment without content memory deficits were not seen in previously reported thalamic amnesic patients. Accordingly, the present patient's poor retrograde temporal memory could hardly be explained by only a thalamic lesion. It is concluded that the patient's impairment of temporal order memory for the retrograde material is probably due to the direct disconnection between the frontal lobe and the hippocampus by disruption of the fornix. PMID- 10369833 TI - Focal (segmental) dyshidrosis in syringomyelia. AB - The features or mechanisms of dyshidrosis have not been sufficiently clarified. Neither has the difference between hyperhidrosis and hypohidrosis. To clarify the features and mechanisms of dyshidrosis (hyperhidrosis and hypohidrosis) in syringomyelia, the clinical features focusing on hidrosis of 30 patients with syringomyelia and Chiari malformation located from a syringomyelia database were prospectively analysed. The patients were classified into three groups: eight patients (26.7%) had segmental hypohidrosis, 10 (33. 3%) had segmental hyperhidrosis, and 12 (40.0%) had normohidrosis. We found that the Karnofsky functional status for the hyperhydrosis and normohidrosis groups were significantly higher than for the hypohidrosis group (p=0.0012), with no significant differences between the hyperhidrosis and normohidrosis groups. The duration from the onset of syringomyelia to the current dyshidrosis was significantly longer in the hypohidrosis group than in the hyperhidrosis group (p=0.0027). A significant correlation was identified between the duration from the onset of syringomyelia to the time at study and the performance score (r= 0.599, p=0.0003). The results substantiate previous hypotheses that in its early stage syringomyelia causes segmental hyperactivity of the sympathetic preganglionic neurons, and hyperactivity of these gradually subsides as tissue damage progresses. Focal hyperhidrosis may be regarded as a hallmark of a relatively intact spinal cord, as well as normohidrosis. PMID- 10369834 TI - Decreased cerebellar blood flow in postinfectious acute cerebellar ataxia. AB - OBJECTIVE: The aim of the present study was to evaluate the regional cerebral blood flow (rCBF) in patients with postinfectious acute cerebellar ataxia using single photon emission computed tomography (SPECT). METHODS: Five children with postinfectious acute cerebellar ataxia and five control subjects were examined. The distribution of rCBF was measured by SPECT imaging after intravenous administration of 123I-IMP (111 MBq). The rCBF ratio-defined as the ratio of rCBF in the region of interest (ROI) to that in the occipital cortex-was calculated for each cortical and subcortical ROI. The mean rCBF ratio of each region was then compared between the ataxic and control subjects. These patients and all control subjects were also evaluated using MRI. RESULTS: The rCBF ratio was significantly lower in the cerebellum of the ataxic patients than in the cerebellum of the control subjects (p<0.05). No abnormal cerebellar morphology and no abnormal signal intensities were found on MRI. CONCLUSION: 123I-IMP SPECT clearly demonstrated the decreased rCBF in the cerebellum of all patients with postinfectious acute cerebellar ataxia. PMID- 10369835 TI - Right frontal lobe slow frequency repetitive transcranial magnetic stimulation (SF r-TMS) is an effective treatment for depression: a case-control pilot study of safety and efficacy. AB - Major depression may result from decreased left frontal lobe function with respect to the right. Fast frequency repetitive transcranial magnetic stimulation (FF r-TMS) excites the underlying cortex whereas slow frequency repetitive transcranial magnetic stimulation (SF r-TMS) causes cortical inhibition. Left frontal FF r-TMS attenuates major depression whereas the inhibitory effects of right frontal SF r-TMS are unknown. This study tested the hypothesis that right frontal SF r-TMS would treat depressed patients with minimal effect on controls. A psychiatrist administered the Beck depression inventory and Hamilton D depression rating scales to eight depressed patients and six controls before and after the treatment protocol. Eight sessions of 100 right frontal lobe SF r-TMS were given at motor threshold and 0.5 Hz over a 6 week period. No adverse outcomes were noted in either group. A significant antidepressant effect was noted in depressed patients on the Beck and Hamilton D depression rating scales (p<0.05). No change on either scale was noted in the controls. In conclusion right frontal lobe SF r-TMS is a safe, non-invasive treatment for major depression that deserves further investigation. PMID- 10369836 TI - Severe necrotising cutaneous lesions complicating treatment with interferon beta 1a. PMID- 10369838 TI - Single thalamic-subthalamic artery and bilateral thalamic infarcts. PMID- 10369837 TI - Autosomal dominant muscle cramp syndrome in a Japanese family. AB - OBJECTIVES: To identify the clinical, electrophysiological, histological, and genetic characteristics of a Japanese family with a muscle cramp syndrome. METHODS: Fourteen patients (eight men, six women) were studied in four generations of a single family. Electrophysiological examinations were performed in four cases and muscle and nerve biopsies were performed on the propositus. RESULTS: The mode of inheritance seemed to be autosomal dominant. The cramps occurred during both exertion and at rest, and during sleep. Electromyographic examination indicated a neurogenic aetiology. There was a decreased number of large myelinated fibres in the sural nerve, and fibre type grouping in the quadriceps femoris muscle biopsy. CONCLUSIONS: The autosomal dominant muscle cramp syndrome in this family is probably caused by a polyneuropathy. PMID- 10369839 TI - Tetanus originating from a benign scalp tumour. PMID- 10369840 TI - Sensory alien hand syndrome. PMID- 10369841 TI - Vasomotor reactivity is exhuasted in transient ischaemic attacks with limb shaking. PMID- 10369842 TI - Calvarial and dural reconstruction PMID- 10369844 TI - Spinal cord diseases-diagnosis and treatment PMID- 10369843 TI - Echoenhancers and transcranial color duplex sonography PMID- 10369845 TI - Neurologic disease in women PMID- 10369846 TI - The rabbit lens epithelial cell line N/N1003A requires 12-lipoxygenase activity for DNA synthesis in response to EGF. AB - PURPOSE: Cultured rat lenses and primary human lens epithelial cells (HLECs) express12-lipoxygenase (12-LOX) and require a 12-LOX metabolite of arachidonic acid for growth in response to EGF and insulin. This study seeks to identify an established cell line with these characteristics. METHODS: Immunoblotting was used to screen eight lens epithelial cell lines for 12-LOX expression: the human line, HLE-B3; mouse lines alphaTN4, 17EM15, 21EM15, and MLE6, and rabbit lines N/N1003A, LEP2 and B3. DNA synthesis was measured as incorporation of 3H thymidine into DNA. Expression of c-fos mRNA was detected by RT-PCR. The involvement of 12-lipoxygenase metabolites was determined using the lipoxygenase inhibitors baicalein, cinnamyl 3,4-dihydroxy-alpha-cyanocinnamate (CDC), or nordihydroguiairetic acid (NDGA). RESULTS: 12-LOX was detected only in the rabbit lines N/N1003A, LEP2 and B3. N/N1003A cells were chosen for further study. 12-LOX inhibitors blocked DNA synthesis in response to EGF with or without insulin. Inhibition of EGF-stimulated DNA synthesis was reversed by 0.3 microM to 3 microM 12(S)hydroxyeicosatetraenoic acid (HETE), but not by equivalent concentrations of 5(S)HETE, 8(S)HETE, 15(S)HETE, or 12(R)HETE. Baicalein prevented EGF induction of c-fos mRNA. The transformed HLEC line, HLE-B3, showed little stimulation of DNA synthesis in response to EGF and was unaffected by the presence of 12-LOX inhibitors. CONCLUSIONS: N/N1003A cells, like primary cultured human lens epithelial cells or neonatal rat lenses, require 12-LOX activity for EGF dependent growth. This line will be useful for studies of the mechanism of action of 12(S)HETE. PMID- 10369847 TI - Facilitating transthoracic cardioversion of atrial fibrillation with ibutilide pretreatment. AB - BACKGROUND: Atrial fibrillation cannot always be converted to sinus rhythm by transthoracic electrical cardioversion. We examined the effect of ibutilide, a class III antiarrhythmic agent, on the energy requirement for atrial defibrillation and assessed the value of this agent in facilitating cardioversion in patients with atrial fibrillation that is resistant to conventional transthoracic cardioversion. METHODS: One hundred patients who had had atrial fibrillation for a mean (+/-SD) of 117+/-201 days were randomly assigned to undergo transthoracic cardioversion with or without pretreatment with 1 mg of ibutilide. We designed a step-up protocol in which shocks at 50, 100, 200, 300, and 360 J were used for transthoracic cardioversion. If transthoracic cardioversion was unsuccessful in a patient who had not received ibutilide pretreatment, ibutilide was administered and transthoracic cardioversion attempted again. RESULTS: Conversion to sinus rhythm occurred in 36 of 50 patients who had not received ibutilide (72 percent) and in all 50 patients who had received ibutilide (100 percent, P<0.001). In all 14 patients in whom transthoracic cardioversion alone failed, sinus rhythm was restored when cardioversion was attempted again after the administration of ibutilide. Pretreatment with ibutilide was associated with a reduction in the mean energy required for defibrillation (166+/-80 J, as compared with 228+/-93 J without pretreatment; P<0.001). Sustained polymorphic ventricular tachycardia occurred in 2 of the 64 patients who received ibutilide (3 percent), both of whom had an ejection fraction of 0.20 or less. The rates of freedom from atrial fibrillation after six months of follow-up were similar in the two randomized groups. CONCLUSIONS: The efficacy of transthoracic cardioversion for converting atrial fibrillation to sinus rhythm was enhanced by pretreatment with ibutilide. However, use of this drug should be avoided in patients with very low ejection fractions. PMID- 10369848 TI - Prevention of implantable-defibrillator shocks by treatment with sotalol. d,l Sotalol Implantable Cardioverter-Defibrillator Study Group. AB - BACKGROUND: Patients with implantable cardioverter-defibrillators often receive adjunctive antiarrhythmic therapy to prevent frequent shocks. We tested the efficacy and safety of sotalol, a beta-blocker with class III antiarrhythmic effects, for this purpose. METHODS: In a multicenter trial, patients were stratified according to left ventricular ejection fraction (< or =0.30 or >0.30), randomly assigned to double-blind treatment with 160 to 320 mg of sotalol per day (151 patients) or matching placebo (151 patients), and followed for 12 months. Kaplan-Meier analyses of the time to an event were performed. Three end points were used: the delivery of a first shock for any reason or death from any cause, the first appropriate shock for a ventricular arrhythmia or death from any cause, and the first inappropriate shock for a supraventricular arrhythmia or death from any cause. RESULTS: Compliance with double-blind treatment was similar in the two groups. There were seven deaths in the placebo group and four in the sotalol group. As compared with placebo, treatment with sotalol was associated with a lower risk of death from any cause or the delivery of a first shock for any reason (reduction in risk, 48 percent; P<0.001 by the log-rank test), death from any cause or the delivery of a first appropriate shock (reduction in risk, 44 percent; P=0.007), or death from any cause or the delivery of a first inappropriate shock (reduction in risk, 64 percent; P=0.004). Sotalol also reduced the mean (+/-SD) frequency of shocks due to any cause (1.43+/-3.53 shocks per year, as compared with 3.89+/-10.65 in the placebo group; P=0.008). In the sotalol group, the reduction in the risk of death from any cause or the delivery of a first shock for any reason did not differ significantly between patients with ejection fractions of more than 0.30 and those with ejection fractions of 0.30 or less. CONCLUSIONS: Oral sotalol was safe and efficacious in reducing the risk of death or the delivery of a first defibrillator shock whether or not ventricular function was depressed. PMID- 10369849 TI - Antibodies against human herpesvirus 8 in black South African patients with cancer. AB - BACKGROUND: Infection with human herpesvirus 8 (HHV-8) has been consistently linked to Kaposi's sarcoma, but its mode of transmission, association with other cancers, and interaction with the human immunodeficiency virus type 1 (HIV-1) are largely unknown. METHODS: Between January 1992 and December 1997, we interviewed 3591 black patients with cancer in Johannesburg and Soweto, South Africa. Blood was tested for antibodies against HIV-1 and HHV-8 in 3344 of the patients. Antibodies against HHV-8 were detected with an indirect immunofluorescence assay. The intensity of the fluorescent signal correlated well with the titers of antibodies (P<0.001). The relations among the presence of anti-HHV-8 antibodies, sociodemographic and behavioral factors, type of cancer, and the presence or absence of coexistent HIV infection were examined with the use of unconditional logistic-regression models. RESULTS: Among the 3293 subjects with cancers other than Kaposi's sarcoma, the standardized seroprevalence of antibodies against HHV 8 was 32 percent, which did not differ significantly from the standardized seroprevalence among black blood donors. Among these 3293 patients, the prevalence of antibodies against HHV-8 increased with increasing age (P<0.001) and an increasing number of sexual partners (P=0.05) and decreased with increasing years of education (P=0.007); it was not strongly associated with HIV 1 infection. Anti-HHV-8 antibodies were more frequent among black than white blood donors (P<0.001). Among the 51 patients with Kaposi's sarcoma, the standardized seroprevalence of antibodies against HHV-8 was 83 percent, significantly higher than the prevalence among those without Kaposi's sarcoma (P<0.001). For 16 other specific types of cancer, including multiple myeloma (108 cases) and prostate cancer (202 cases), the variation in the standardized seroprevalence of antibodies against HHV-8 was not remarkable. At a given intensity of fluorescence of anti-HHV-8 antibodies, Kaposi's sarcoma was more frequent among HIV-1-positive patients than among those who were HIV-1-negative (P<0.001). CONCLUSIONS: Among black patients with cancer in South Africa, the seroprevalence of anti-HHV-8 antibodies is high and is specifically associated with Kaposi's sarcoma, particularly at high titers. PMID- 10369850 TI - Plasma normetanephrine and metanephrine for detecting pheochromocytoma in von Hippel-Lindau disease and multiple endocrine neoplasia type 2. AB - BACKGROUND: The detection of pheochromocytomas in patients at risk for these tumors, such as patients with von Hippel-Lindau disease or multiple endocrine neoplasia type 2 (MEN-2), is hindered by the inadequate sensitivity of commonly available biochemical tests. In this study we evaluated measurements of plasma normetanephrine and metanephrine for detecting pheochromocytomas in patients with von Hippel-Lindau disease or MEN-2. METHODS: We studied 26 patients with von Hippel-Lindau disease and 9 patients with MEN-2 who had histologically verified pheochromocytomas and 50 patients with von Hippel-Lindau disease or MEN-2 who had no radiologic evidence of pheochromocytoma. Von Hippel-Lindau disease and MEN-2 were diagnosed on the basis of germ-line mutations of the appropriate genes. The plasma concentrations of normetanephrine and metanephrine were compared with the plasma concentrations of catecholamines (norepinephrine and epinephrine) and urinary excretion of catecholamines, metanephrines, and vanillylmandelic acid. RESULTS: The sensitivity of measurements of plasma normetanephrine and metanephrine for the detection of tumors was 97 percent, whereas the other biochemical tests had a sensitivity of only 47 to 74 percent. All patients with MEN-2 had high plasma concentrations of metanephrine, whereas the patients with von Hippel-Lindau disease had almost exclusively high plasma concentrations of only normetanephrine. One patient with von Hippel-Lindau disease had a normal plasma normetanephrine concentration; this patient had a very small adrenal tumor (<1 cm). The high sensitivity of measurements of plasma normetanephrine and metanephrine was accompanied by a high level of specificity (96 percent). CONCLUSIONS: Measurements of plasma normetanephrine and metanephrine are useful in screening for pheochromocytomas in patients with a familial predisposition to these tumors. PMID- 10369851 TI - Images in clinical medicine. von Hippel-Lindau disease. PMID- 10369853 TI - Gastrointestinal toxicity of nonsteroidal antiinflammatory drugs. PMID- 10369852 TI - The relation between funding by the National Institutes of Health and the burden of disease. AB - BACKGROUND: The Institute of Medicine has proposed that the amount of disease specific research funding provided by the National Institutes of Health (NIH) be systematically and consistently compared with the burden of disease for society. METHODS: We performed a cross-sectional study comparing estimates of disease specific funding in 1996 with data on six measures of the burden of disease. The measures were total mortality, years of life lost, and number of hospital days in 1994 and incidence, prevalence, and disability-adjusted life-years (one disability-adjusted life-year is defined as the loss of one year of healthy life to disease) in 1990. With the use of these measures as explanatory variables in a regression analysis, predicted funding was calculated and compared with actual funding. RESULTS: There was no relation between the amount of NIH funding and the incidence, prevalence, or number of hospital days attributed to each condition or disease (P=0.82, P=0.23, and P=0.21, respectively). The numbers of deaths (r=0.40, P=0.03) and years of life lost (r=0.42, P=0.02) were weakly associated with funding, whereas the number of disability-adjusted life-years was strongly predictive of funding (r=0.62, P<0.001). When the latter three measures were used to predict expected funding, the conclusions about the appropriateness of funding for some diseases varied according to the measure used. However, the acquired immunodeficiency syndrome, breast cancer, diabetes mellitus, and dementia all received relatively generous funding, regardless of which measure was used as the basis for calculating support. Research on chronic obstructive pulmonary disease, perinatal conditions, and peptic ulcer was relatively underfunded. CONCLUSIONS: The amount of NIH funding for research on a disease is associated with the burden of the disease; however, different measures of the burden of disease may yield different conclusions about the appropriateness of disease-specific funding levels. PMID- 10369854 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 18-1999. A 54-year-old woman with acute renal failure and thrombocytopenia. PMID- 10369855 TI - Redefining the role of antiarrhythmic drugs. PMID- 10369856 TI - Human herpesvirus 8 and Kaposi's sarcoma--some answers, more questions. PMID- 10369857 TI - Evaluating the burden of disease and spending the research dollars of the National Institutes of Health. PMID- 10369858 TI - Health care reform at the close of the 20th century. PMID- 10369859 TI - Defective chromosome segregation, microtubule bundling and nuclear bridging in inner centromere protein gene (Incenp)-disrupted mice. AB - INCENP is a chromosomal passenger protein which relocates from the centromere to thel spindle midzone during the metaphase-anaphase transition, ultimately being discarded in the cell midbody at the completion of cytokinesis. Using homologous recombination, we have generated Incenp gene-targeted heterozygous mice that are phenotypically indistinguishable from their wild-type littermates. Intercrossing the hetero-zygotes results in no live-born homozygous Incenp -disrupted progeny, indicating an early lethality. Day 3.5 affected pre-implantation embryos contain large, morphologically abnormal cells that fail to fully develop a blastocoel cavity or thrive in utero and in culture. Chromatin and tubulin immunocytochemical stainings of these and day 2.5 affected embryos reveal a high mitotic index, no discernible metaphase or anaphase stages, complete absence of midbodies, micronuclei formation, morphologically irregular macronuclei with large chromosome complements, multipolar mitotic configurations, binucleated cells, internuclear bridges and abnormal spindle bundling. The phenotype is consistent with a defect in the modulation of microtubule dynamics, severely affecting chromosome segregation and resulting in poorly resolved chromatin masses, aberrant karyokinesis and internuclear bridge formation. These latter occurrences could pose a physical barrier blocking cytokinesis. PMID- 10369860 TI - A common molecular basis for rearrangement disorders on chromosome 22q11. AB - The chromosome 22q11 region is susceptible to rearrangements that are associated with congenital anomaly disorders and malignant tumors. Three congenital anomaly disorders, cat-eye syndrome, der() syndrome and velo-cardio-facial syndrome/DiGeorge syndrome (VCFS/DGS) are associated with tetrasomy, trisomy or monosomy, respectively, for part of chromosome 22q11. VCFS/DGS is the most common syndrome associated with 22q11 rearrangements. In order to determine whether there are particular regions on 22q11 that are prone to rearrangements, the deletion end-points in a large number of VCFS/DGS patients were defined by haplotype analysis. Most VCFS/DGS patients have a similar 3 Mb deletion, some have a nested distal deletion breakpoint resulting in a 1.5 Mb deletion and a few rare patients have unique deletions or translocations. The high prevalence of the disorder in the population and the fact that most cases occur sporadically suggest that sequences at or near the breakpoints confer susceptibility to chromosome rearrangements. To investigate this hypothesis, we developed hamster human somatic hybrid cell lines from VCFS/DGS patients with all three classes of deletions and we now show that the breakpoints occur within similar low copy repeats, termed LCR22s. To support this idea further, we identified a family that carries an interstitial duplication of the same 3 Mb region that is deleted in VCFS/DGS patients. We present models to explain how the LCR22s can mediate different homologous recombination events, thereby generating a number of rearrangements that are associated with congenital anomaly disorders. We identified five additional copies of the LCR22 on 22q11 that may mediate other rearrangements leading to disease. PMID- 10369861 TI - High level expression of expanded full-length ataxin-3 in vitro causes cell death and formation of intranuclear inclusions in neuronal cells. AB - Spinocerebellar ataxia type 3 (SCA3) is caused by a CAG/polyglutamine repeat expansion in the SCA3 gene. To analyse the pathogenic mechanisms in SCA3, we have generated ataxin-3-expressing rat mesencephalic CSM14.1 cells. In these cells, a post-mitotic neuronal phenotype is induced by temperature shift. The isolated stable cell lines provided high level expression of non-expanded (Q23) or expanded (Q70) human full-length ataxin-3. CSM14.1 cells expressing the expanded full-length ataxin-3 developed nuclear inclusion bodies, strong indentations of the nuclear envelope and cytoplasmic vacuolation. These ultrastructural alterations were present prior to a significantly decreased viability of neuronally differentiated cells expressing expanded ataxin-3. The observed spontaneous cell death did not correlate with formation of intranuclear inclusions and was not apoptotic by ultrastructural analysis. No increased susceptibility to staurosporine-induced apoptosis was found for the expanded or non-expanded ataxin-3-expressing cell lines. These data show that high level expression of expanded full-length ataxin-3 in a neuron-like cell line generates ultrastructural alterations of SCA3 pathogenesis and results in increased spontaneous non-apoptotic cell death. PMID- 10369862 TI - A single nucleotide difference that alters splicing patterns distinguishes the SMA gene SMN1 from the copy gene SMN2. AB - Spinal muscular atrophy (SMA) is a recessive disorder characterized by loss of motor neurons in the spinal cord. It is caused by mutations in the telomeric survival motor neuron 1 ( SMN1 ) gene. Alterations within an almost identical copy gene, the centromeric survival motor neuron 2 ( SMN2 ) gene produce no known phenotypic effect. The exons of the two genes differ by just two nucleotides, neither of which alters the encoded amino acids. At the genomic level, only five nucleotides that differentiate the two genes from one another have been reported. The entire genomic sequence of the two genes has not been determined. Thus, differences which might explain why SMN1 is the SMA gene are not readily apparent. In this study, we have completely sequenced and compared genomic clones containing the SMN genes. The two genes show striking similarity, with the homology being unprecedented between two different yet functional genes. The only critical difference in an approximately 32 kb region between the two SMN genes is the C->T base change 6 bp inside exon 7. This alteration but not other variations in the SMN genes affects the splicing pattern of the genes. The majority of the transcript from the SMN1 locus is full length, whereas the majority of the transcript produced by the SMN2 locus lacks exon 7. We suggest that the exon 7 nucleotide change affects the activity of an exon splice enhancer. In SMA patients, the loss of SMN1 but the presence of SMN2 results in low levels of full length SMN transcript and therefore low SMN protein levels which causes SMA. PMID- 10369863 TI - Abundant expression and cytoplasmic aggregations of [alpha]1A voltage-dependent calcium channel protein associated with neurodegeneration in spinocerebellar ataxia type 6. AB - Spinocerebellar ataxia type 6 (SCA6) is one of the eight neurodegenerative diseases caused by a tri-nucleotide (CAG) repeat expansion coding polyglutamine (CAG repeat/polyglutamine diseases) and is characterized by late onset autosomal dominant cerebellar ataxia and predominant loss of cerebellar Purkinje cells. Although the causative, small and stable CAG repeat expansion for this disease has been identified in the [alpha]1A voltage-dependent calcium channel gene (CACNA1A), the mechanism which leads to predominant Purkinje cell degeneration is totally unknown. In this study, we show that the calcium channel mRNA/protein containing the CAG repeat/polyglutamine tract is most intensely expressed in Purkinje cells of human brains. In SCA6 brains, numerous oval or rod-shaped aggregates were seen exclusively in the cytoplasm of Purkinje cells. These cytoplasmic inclusions were not ubiquitinated, which contrasts with the neuronal intra-nuclear inclusions of other CAG repeat/polyglutamine diseases. In cultured cells, formation of perinuclear aggregates of the channel protein and apoptotic cell death were seen when transfected with full-length CACNA1A coding an expanded polyglutamine tract. The present study indicates that the mechanism of neurodegeneration in SCA6 is associated with cytoplasmic aggregations of the [alpha]1A calcium channel protein caused by a small CAG repeat/polyglutamine expansion in CACNA1A. PMID- 10369864 TI - The cytotoxic T lymphocyte antigen-4 is a major Graves' disease locus. AB - Graves' disease (GD) is an autoimmune thyroid disorder that is inherited as a complex trait. We have genotyped 77 affected sib-pairs with autoimmune thyroid disease for eight polymorphic markers spanning the cytotoxic T lymphocyte antigen 4 ( CTLA-4 ) region of chromosome 2q31-q33, and for five markers spanning the major histocompatibility complex ( MHC ) region of chromosome 6p21. Non parametric analysis showed linkage of GD to the CTLA-4 region with a peak non parametric linkage (NPL) score of 3.43 ( P = 0.0004) at the marker D2S117. The proportion of affected full-sibs sharing zero alleles (z0) reached a minimum of 0.113 close to D2S117, giving a locus-specific lambdas for this region of 2.2. Families with brother-sister sib-pairs showed a peak NPL of 3.46 ( P = 0.0003, lambdas > 10) at D2S117, compared with 2.00 ( P = 0.02, lambdas = 1.9) in the families with only affected females, suggesting a stronger influence in families with affected males. Association between GD and the G allele of the Thr17Ala polymorphism within the CTLA-4 gene ( CTLA4A/G ) was observed using unaffected sib controls ( P = 0.005). Lesser evidence for linkage was found at the MHC locus, with a peak NPL score of 1.95 ( P = 0.026), between the markers D6S273 and TNFalpha. We demonstrate that the CTLA-4 locus (lambdas = 2.2) and the MHC locus (lambdas = 1.6) together confer approximately 50% of the inherited susceptibility to GD disease in our population. PMID- 10369865 TI - New gene family defined by MORC, a nuclear protein required for mouse spermatogenesis. AB - Mammalian spermatogenesis is a complex developmental process. The analysis of mouse mutations has provided insight into biochemical pathways required for completion of this process. We previously described the autosomal recessive mouse morc TgN(Tyr)1Az(microrchidia) mutation, a serendipitous transgenic insertional mutation which causes arrest of spermatogenesis prior to the pachytene stage of meiosis prophase I. We now report the molecular characterization of the morc locus and positional cloning of a gene disrupted by the morc TgN(Tyr)1Az mutation. This gene, which we term Morc, encodes a 108 kDa protein expressed specifically in male germ cells. The transgene integrated within the first intron of Morc and was accompanied by an intragenic deletion of approximately 13 kb of genomic sequences, removing exons 2-4 and abrogating expression of the wild-type transcript. Analysis of the MORC protein sequence revealed putative nuclear localization signals, two predicted coiled-coil structural motifs and limited homology to GHL (GyraseB, Hsp90, MutL) ATPase. Epitope-tagged MORC protein expressed in COS7 cells localized to the nucleus. We also cloned the human MORC homolog and show that it too is testis-specific, but closely related human genes are transcribed in multiple somatic tissues. Homologous proteins are also present in zebrafish, nematodes, slime mold and plants. Thus, cloning of Morc defines a novel gene family whose members are likely to serve important biological functions in both meiotic and mitotic cells of multicellular organisms. PMID- 10369866 TI - LIT1, an imprinted antisense RNA in the human KvLQT1 locus identified by screening for differentially expressed transcripts using monochromosomal hybrids. AB - Mammalian imprinted genes are frequently arranged in clusters on particular chromosomes. The imprinting cluster on human chromosome 11p15 is associated with Beckwith-Wiedemann syndrome (BWS) and a variety of human cancers. To clarify the genomic organization of the imprinted cluster, an extensive screen for differentially expressed transcripts in the 11p15 region was performed using monochromosomal hybrids with a paternal or maternal human chromosome 11. Here we describe an imprinted antisense transcript identified within the KvLQT1 locus, which is associated with multiple balanced chromosomal rearrangements in BWS and an additional breakpoint in embryonal rhabdoid tumors. The transcript, called LIT1 (long QT intronic transcript 1), was expressed preferentially from the paternal allele and produced in most human tissues. Methylation analysis revealed that an intronic CpG island was specifically methylated on the silent maternal allele and that four of 13 BWS patients showed complete loss of maternal methylation at the CpG island, suggesting that antisense regulation is involved in the development of human disease. In addition, we found that eight of eight Wilms' tumors exhibited normal imprinting of LIT1 and five of five tumors displayed normal differential methylation at the intronic CpG island. This contrasts with five of six tumors showing loss of imprinting of IGF2. We conclude that the imprinted gene domain at the KvLQT1 locus is discordantly regulated in cancer from the imprinted domain at the IGF2 locus. Thus, this positional approach using human monochromosomal hybrids could contribute to the efficient identification of imprinted loci in humans. PMID- 10369867 TI - Identification of survival motor neuron as a transcriptional activator-binding protein. AB - Spinal muscular atrophy (SMA) is an inherited neuro-muscular disease characterized by specific degeneration of spinal cord anterior horn cells and subsequent muscle atrophy. Survival motor neuron ( SMN ), located on chromosome 5q13, is the SMA-determining gene. In the nucleus, SMN is present in large foci called gems, the function of which is not yet known, while cytoplasmic SMN has been implicated in snRNP biogenesis. In SMA patients, SMN protein levels and the number of gems generally correlate with disease severity, suggesting a critical nuclear function for SMN. In a screen for proteins associated with the nuclear transcription activator 'E2' of papillomavirus, two independent SMN cDNAs were isolated. The E2 and SMN proteins were found to associate specifically in vitro and in vivo. Expression of SMN enhanced E2-dependent transcriptional activation, and patient-derived SMN missense mutations reduced E2 gene expression. Our results demonstrate that SMN interacts with a nuclear transcription factor and imply that SMN may serve a role in regulating gene expression. These observations suggest that SMA may in part result from abnormal gene expression and that E2 may influence viral gene expression through SMN interaction. PMID- 10369868 TI - Ultrastructural localization and progressive formation of neuropil aggregates in Huntington's disease transgenic mice. AB - How aggregates of polyglutamine proteins are involved in the neurological symptoms of glutamine repeat diseases is unknown. We show that huntingtin aggregates are present in the neuronal processes of transgenic mice that express exon 1 of the Huntington's disease (HD) gene. Unlike aggregates in the nucleus, these neuropil aggregates are usually smaller and are not ubiquitinated. Electron microscopy reveals many neuropil aggregates in axons and axon terminals. Huntingtin aggregates in the axon terminal are co-localized with some synaptic vesicles, implying that they may affect synaptic transmission and neuronal communication. The formation of neuropil aggregates is highly correlated with the development of neurological symptoms. The present study raises the possibility that neuropil aggregates may cause a dysfunction in neuronal communication and con-tribute to the neurological symptoms of HD. PMID- 10369869 TI - A missense mutation in connexin26, D66H, causes mutilating keratoderma with sensorineural deafness (Vohwinkel's syndrome) in three unrelated families. AB - The multiplicity of functions served by intercellular gap junctions is reflected by the variety of phenotypes caused by mutations in the connexins of which they are composed. Mutations in the connexin26 (Cx26) gene ( GJB2 ) at 13q11-q13 are a major cause of autosomal recessive hearing loss (DFNB1), but have also been reported in autosomal dominant deafness (DFNA3). We now report a Cx26 mutation in three families with mutilating keratoderma and deafness [Vohwinkel's syndrome (VS; MIM 124500), as originally described]. VS is characterized by papular and honeycomb keratoderma associated with constrictions of digits leading to autoamputation, distinctive starfish-like acral keratoses and moderate degrees of deafness. In a large British pedigree, we have mapped the defect to the Cx26 locus. All 10 affected members were heterozygous for a non-conservative mutation, D66H, in Cx26. The same mutation was found subsequently in affected individuals from two unrelated Spanish and Italian pedigrees segregating VS, suggesting that D66H in Cx26 is a common mutation in classical VS. This mutation occurs at a highly conserved residue in the first extracellular domain of the Cx26 molecule, and may exert its effects by interfering with assembly into connexons, docking with adjacent cells or gating properties of the gap junction. Our results provide evidence that a specific mutation in Cx26 can impair epidermal differentiation, as well as inner ear function. PMID- 10369870 TI - Functional consequences of mutations in the early growth response 2 gene (EGR2) correlate with severity of human myelinopathies. AB - The early growth response 2 gene ( EGR2 ) is a Cys2His2zinc finger transcription factor which is thought to play a role in the regulation of peripheral nervous system myelination. This idea is based partly on the phenotype of homozygous Krox20 ( Egr2 ) knockout mice, which display hypomyelination of the PNS and a block of Schwann cells at an early stage of differentiation. Mutations in the human EGR2 gene have recently been associated with the inherited peripheral neuropathies Charcot-Marie-Tooth type 1, Dejerine-Sottas syndrome and congenital hypomyelinating neuropathy. Three of the four EGR2 mutations are dominant and occur within the zinc finger DNA-binding domain. The fourth mutation is recessive and affects the inhibitory domain (R1) that binds the NAB transcriptional co repressors. A combination of DNA-binding assays and transcriptional analysis was used to determine the functional consequences of these mutations. The zinc finger mutations affect DNA binding and the amount of residual binding directly correlates with disease severity. The R1 domain mutation prevents interaction of EGR2 with the NAB co-repressors and thereby increases transcriptional activity. These data provide insight into the possible disease mechanisms underlying EGR2 mutations and the reason for varying severity and differences in inheritance patterns. PMID- 10369871 TI - A complex pattern of evolutionary conservation and alternative polyadenylation within the long 3"-untranslated region of the methyl-CpG-binding protein 2 gene (MeCP2) suggests a regulatory role in gene expression. AB - A systematic search for expressed sequences in the human Xq28 region resulted in the isolation of 8.5 kb large contigs of human and murine cDNAs with no apparent conserved open reading frames. These cDNAs were found to be derived from the 3" untranslated region (3"-UTR) of the methyl-CpG-binding protein 2 gene ( MeCP2 ). This long 3"-UTR is part of an alternatively polyadenylated, 10.1 kb MeCP2 transcript which is differentially expressed in human brain and other tissues. RNA in situ hybridization to sections of mouse embryo and adult tissues of an Mecp2 3"-UTR probe showed ubiquitous low level expression in early organogenesis and enhanced expression in the hippocampus during formation of the differentiated brain. Sequence comparison between the human and mouse homologues revealed several blocks of very high conservation separated by less conserved sequences. Additional support for a domain-like conservation pattern of the long 3"-UTR of the MeCP2 gene was obtained by examining conservation in the chimpanzee, orangutan, macaque, hamster, rat and kangaroo. The minimum free energy distribution for the predicted RNA secondary structure was very similar in human and mouse sequences. In particular, the conserved blocks were predicted to be of high minimum free energy, which suggests weak secondary structure with respect to RNA folding. The fact that both the sequence and predicted secondary structure have been highly conserved during evolution suggests that both the primary sequence and the three-dimensional structure of the 3"-UTR may be important for its function in post-transcriptional regulation of MeCP2 expression. PMID- 10369872 TI - Presenilins interact with Rab11, a small GTPase involved in the regulation of vesicular transport. AB - Presenilin 1 (PS1) mutations account for the majority of early-onset dominant cases of familial Alzheimer's disease. Presenilins (PSs) are located in many intra-cellular compartments such as the endoplasmic reticulum, Golgi apparatus, nuclear region and vesicular structures. These proteins include from seven to nine putative transmembrane domains, with the N- and C-terminal ends and a large hydrophilic loop orientated towards the cytoplasm. We report an interaction between the human PS1 or PS2 hydrophilic loop and Rab11, a small GTPase belonging to the Ras-related superfamily. Interaction domains were mapped to codons 374-400 for PS1 and to codons 106-179 for Rab11, a region including the fourth GTP binding domain. Considering the implication of Rab proteins in vesicular transport pathways, the PS-Rab11 inter-action suggests that PSs might be involved in amyloid precursor protein vesicular routing. PMID- 10369873 TI - Up71 and up140, two novel transcripts of utrophin that are homologues of short forms of dystrophin. AB - Utrophin is a large protein which accumulates at the neuromuscular synapse and myotendinous junctions in adult skeletal muscle, and is widely expressed in several non-skeletal muscle tissues. Evidence from a variety of sources suggests that a successful strategy for treatment of Duchenne muscular dystrophy patients will be to increase expression of utrophin in muscle. There is still much to be learnt about utrophin gene regulation, in particular regarding alternative isoforms, their promoters and role in muscle and non-muscle tissues. Using 5" RACE we have identified two novel transcripts of utrophin, Up71 and Up140, with unique first exons and promoters located in intron 62 and intron 44, respectively. These transcripts appear to be structural homologues of the short dystrophin transcripts, Dp140 and Dp71, emphasizing the high degree of structural conservation between the utrophin and dystrophin genes. RT-PCR shows that Up71 and Up140 are widely expressed in both human and mouse tissues, including skeletal muscle. We present evidence for transcript-specific differential mRNA splicing of exon 71, in both Up71 and Up140, similar to that described for dystrophin. No evidence for splicing of exon 78 of utrophin was found. This is in contrast to dystrophin and may reflect a subtle functional difference in patterns of phosphorylation between the two proteins. PMID- 10369874 TI - Allelic and locus heterogeneity in inherited venous malformations. AB - Venous malformations are low-flow vascular lesions consisting of disorganized thin-walled vascular channels. These can occur sporadically but also as an autosomal dominant condition termed venous malformations, cutaneous and mucosal (VMCM; OMIM 600195). In two large unrelated kindreds mapping to chromosome 9, the identical R849W missense mutation was identified in the first kinase domain of Tie2, an endothelial cell-specific receptor tyrosine kinase. We report here the identification of four new kindreds with inherited venous malformations. Unlike the initial two families described, these four families demonstrate allelic and locus heterogeneity. In one of these families, the R849W mutation co-segregates with the disease phenotype. Three other families with venous malformations lack this mutation. One of these families is linked to markers near TIE2 on chromosome 9. In this family, we identified a novel mutation within the first kinase domain of Tie2 resulting in a Y897S change. Results from COS-1 cell transfections using expression constructs containing either the R849W or the Y897S mutation suggest that the receptors containing either mutation show ligand-independent hyperphosphorylation. These results suggest a gain-of-function mechanism for development of venous malformations in these families. Of the two remaining families, one excludes linkage to the TIE2 locus, establishing the existence of at least one additional locus for dominantly inherited venous malformations. PMID- 10369875 TI - Expression of human F8B, a gene nested within the coagulation factor VIII gene, produces multiple eye defects and developmental alterations in chimeric and transgenic mice. AB - Factor VIII-associated gene B ( F8B ) is a small human gene of unknown function which is nested within the gene encoding coagulation factor VIII ( FVIII ) in chromosome band Xq28. The sequence of F8B includes the C2 cell adhesion motif of factor VIII, which has also been identified in numerous proteins known to play important roles during development. Here we have constructed both chimeric and transgenic mice expressing normal human F8B to investigate its possible developmental effects. The chimeras produced from embryonic stem cells transfected with normal F8B under control of a cytomegalovirus promoter and selected for neomycin resistance expressed readily detectable levels of F8B mRNA in multiple tissues. They showed growth retardation, microcephaly, reduced longevity and severe ocular defects, and although they were fertile, gave birth to no F8B heterozygous pups. Seven transgenic mouse lines, produced by injection of the transgene into fertilized oocytes, were viable and of normal size but expressed lower levels of F8B mRNA. Strikingly, they showed the same severe eye abnormalities as the chimeras. These defects included anterior segment dysgenesis, absent or abnormal lens, persistence of the primary vitreous, Harderian gland tumors and ectopic pigmented cells, suggesting that migration of neural crest cells might have been perturbed during eye development. In addition, dysplastic retinas and the absence of photoreceptors were observed, providing a mouse model for retinal degeneration. PMID- 10369876 TI - Autosomal recessive familial neurohypophyseal diabetes insipidus with continued secretion of mutant weakly active vasopressin. AB - Familial neurohypophyseal diabetes insipidus is an autosomal dominant disorder characterized by post-natal development of arginine vasopressin (AVP) deficiency due to mutations in the AVP gene. All published mutations affect the signal peptide or the neurophysin-II carrier protein and are presumed to interfere with processing of the preprohormone, leading to neuronal damage. We studied an unusual Palestinian family consisting of asymptomatic first cousin parents and three children affected with neurohypophyseal diabetes insipidus, suggesting autosomal recessive inheritance. All three affected children were homozygous and the parents heterozygous for a single novel mutation (C301->T) in exon 1, replacing Pro7 of mature AVP with Leu (Leu-AVP). Leu-AVP was a weak agonist with approximately 30-fold reduced binding to the human V2 receptor. Measured by radioimmunoassay with a synthetic Leu-AVP standard, serum Leu-AVP levels were elevated in all three children and further increased during water deprivation to as high as 30 times normal. The youngest child (2 years old) was only mildly affected but had Leu-AVP levels similar to her severely affected 8-year-old brother, suggesting that unknown mechanisms may partially compensate for a deficiency of active AVP in very young children. PMID- 10369878 TI - Extensive gene order differences within regions of conserved synteny between the Fugu and human genomes: implications for chromosomal evolution and the cloning of disease genes. AB - The suitability of the Fugu genome to facilitate the identification of candidate human disease genes using comparative positional cloning is dependent upon the extent to which synteny and gene order are conserved between the two species. We have cloned seven Fugu genes which are closely linked to Surfeit genes in two regions of the Fugu genome and have mapped and ordered their human homologues both by PCR analysis of the Genebridge 4 panel of radiation hybrids and by fluorescence in situ hybridization. All seven human genes map to a 3 Mb region of chromosome band 9q34.1, approximately 2-4 Mb proximal to the human Surfeit genes. Although both Fugu regions are syntenic with human chromosome band 9q34, the relative order of the genes differs greatly in the two species. Indeed, some of the genes that are adjacent in the Fugu genome are separated by at least 2-4 Mb in the human genome. This suggests that intra-chromosomal rearrangements, most probably inversions, have been common during the 900 million years of divergent evolution separating Fugu and human. The utility of Fugu to facilitate human disease gene identification by comparative positional cloning is questioned in light of these results. PMID- 10369877 TI - Structural and functional rescue of murine rod photoreceptors by human rhodopsin transgene. AB - Mice carrying a targeted disruption of the rhodopsin gene develop a severe degenerative retinopathy, failing to elaborate rod photoreceptor outer segments (ROS), having no recordable rod electroretinogram (ERG) and losing all of their rod cells over a period of approximately 12 weeks. Murine and human rhodopsins differ in their amino acid sequences. Whether, or to what extent, such variability might influence the ability of human rhodopsin to serve as an adequate structural and functional substitute for the endogenous protein in mouse rod cells bears direct relevance to exploiting the full utility of Rho-/-animals as a model of degenerative retinal disease in man. We crossed Rho-/-mice with mice expressing a wild-type human rhodopsin transgene at levels approximating to those of the endogenous protein. Immunohistological examination of retinal selections from such animals demonstrated ROS of normal number and length and temporal expression of rhodopsin similar to that observed in wild-type animals; that is, immunoreactivity to an anti-rhodopsin antibody became clearly evident by day 3 post-partum. Whereas Rho-/-mice never display a rod ERG response, and even lose cone responses by 12 weeks of age, rescued mice showed 75% normal maximum amplitudes and had ERG b-wave thresholds (based on a 50 microV criterion) within 0.1 log unit of normal wild-type at 20 weeks, and cone amplitudes remained normal at this age. These data demonstrate very substantial structural and functional rescue of the rod photoreceptors of Rho-/-mice and long-term preservation by the human rhodopsin transgene. PMID- 10369879 TI - Mutations in the KCNQ4 gene are responsible for autosomal dominant deafness in four DFNA2 families. AB - We have previously found linkage to chromosome 1p34 in five large families with autosomal dominant non-syndromic hearing impairment (DFNA2). In all five families, the connexin31 gene ( GJB3 ), located at 1p34 and responsible for non syndromic autosomal dominant hearing loss in two small Chinese families, has been excluded as the responsible gene. Recently, a fourth member of the KCNQ branch of the K+channel family, KCNQ4, has been cloned. KCNQ4 was mapped to chromosome 1p34 and a single mutation was found in three patients from a small French family with non-syndromic autosomal dominant hearing loss. In this study, we have analysed the KCNQ4 gene for mutations in our five DFNA2 families. Missense mutations altering conserved amino acids were found in three families and an inactivating deletion was present in a fourth family. No KCNQ4 mutation could be found in a single DFNA2 family of Indonesian origin. These results indicate that at least two and possibly three genes responsible for hearing impairment are located close together on chromosome 1p34 and suggest that KCNQ4 mutations may be a relatively frequent cause of autosomal dominant hearing loss. PMID- 10369880 TI - Myotilin, a novel sarcomeric protein with two Ig-like domains, is encoded by a candidate gene for limb-girdle muscular dystrophy. AB - The striated muscle sarcomeres are highly organized structures composed of actin (thin) and myosin (thick) filaments that slide past each other during contraction. The integrity of sarcomeres is controlled by a set of structural proteins, among which are titin, a giant molecule that contains several immunoglobulin (Ig)-like domains and associates with thin and thick filaments, and [alpha]-actinin, an actin cross-linking protein. Mutations in several sarcomeric and sarcolemmal proteins have been shown to result in muscular dystrophy and cardiomyopathy. On the other hand, the disease genes underlying several disease forms remain to be identified. Here we describe a novel 57 kDa cytoskeletal protein, myotilin. Its N-terminal sequence is unique, but the C terminal half contains two Ig-like domains homologous to titin. Myotilin is expressed in skeletal and cardiac muscle, it co-localizes with [alpha]-actinin in the sarcomeric I--bands and directly interacts with [alpha]-actinin. The human myotilin gene maps to chromosome 5q31 between markers AFM350yB1 and D5S500. The locus of a dominantly inherited limb-girdle muscular dystrophy (LGMD1A) resides in an overlapping narrow segment, and a new type of distal myopathy with vocal cord and pharyngeal weakness (VCPMD) has been mapped to the same locus. The muscle specificity and apparent role as a sarcomeric structural protein raise the possibility that defects in the myotilin gene may cause muscular dystrophy. PMID- 10369881 TI - Multipoint analysis of human chromosome 11p15/mouse distal chromosome 7: inclusion of H19/IGF2 in the minimal WT2 region, gene specificity of H19 silencing in Wilms' tumorigenesis and methylation hyper-dependence of H19 imprinting. AB - WT2 is defined by maternal-specific loss of heterozygosity (LOH) on chromosome 11p15.5 in Wilms' tumors (WTs). The imprinted H19 gene, in this region, is silenced and hypermethylated in most WTs, and this is linked to pathological biallelic expression of IGF2. However, H19 and IGF2 lie within a larger imprinted domain, and the gene specificity of H19 epimutation has been a persistent question. To address this, we assessed LOH, gene expression and DNA methylation at multiple sites in and around the imprinted domain. LOH mapping showed that the entire domain, including IGF2/H19, is within the minimal WT2 region. Genes within the domain, including IPL/TSSC3/BWR1C, IMPT1/ORCTL2/BWR1A/TSSC5, KvLQT1/KCNA9 and TAPA1/CD81, as well as the zinc finger gene ZNF195/ZNFP104 near the centromeric border, were expressed persistently in many WTs. DNA hypermethylation was not detected with 5" upstream probes for IPL, IMPT1, KvLQT1 and ZNF195 in WTs or WT associated kidneys. Fully developed WTs showed variable hypomethylation at an imprinted CpG island in a KvLQT1 intron, but this was only complete in the cases with LOH and was not observed in pre-neoplastic WT-associated kidneys with H19 epimutation. Analysis of the corresponding region of mouse chromosome 7 using methyltransferase-hypomorphic mice showed that the H19 imprint was fully erased, but that the allelic bias at Ipl, Impt1, p57 Kip2 and, to a lesser extent, Kvlqt1, persisted. Pre-existing massive allelic asymmetry for DNA methylation and hyper-dependence of transcription on methylation status may underlie the mechanism of gene-specific silencing of H19 in Wilms' tumorigenesis. PMID- 10369882 TI - Murine modelling of classical lissencephaly. AB - Classical lissencephaly is a severe human neuronal migration disorder characterized by a smooth cerebral surface and a paucity of gyri. Isolated lissencephaly sequence (ILS, OMIM 601545) and Miller-Dieker syndrome (MDS, OMIM 247200) are human malformation syndromes characterized by classical lissencephaly. MDS and some cases of ILS are caused by haploinsufficiency at chromosome 17p13.3. Recent evidence suggests that mutations or deletions of the LIS1 gene, within band 17p13.3, are responsible for classical lissencephaly. LIS1 codes for a subunit of platelet-activating factor acetylhydrolase isoform 1b (PAFAH1B1 or LIS1). To investigate the pathophysiological mechanisms responsible for these two developmental defects, we have undertaken strategies to model these neuronal migration disorders in the mouse. We present a brief review of MDS and ILS, several mouse mutants with cortical neuronal migration defects, and our strategies to model ILS and MDS in the mouse. PMID- 10369884 TI - Evidence that allelic variants of the spinocerebellar ataxia type 2 gene influence susceptibility to multiple sclerosis. AB - Expanded CAG trinucleotide repeats are known to be responsible for five of the autosomal dominant spinocerebellar ataxias (SCA1, SCA2, SCA3, SCA6, and SCA7). We have typed each of these repeats in 226 multiple sclerosis sibling pair families. No expanded repeats were seen, indicating an absence of SCA phenocopies in clinically defined familial multiple sclerosis. However, transmission disequilibrium testing for these repeats demonstrated significant excess transmission of the 22 repeat length allele of the SCA2 gene (P=4. 4E-06) in multiple sclerosis patients. This observation is consistent with pleiotropic effects of the SCA2 gene, with a non-dynamic mutation/polymorphism contributing epistatically to susceptibility in multiple sclerosis. PMID- 10369883 TI - Dystrobrevin- and dystrophin-like mutants display similar phenotypes in the nematode Caenorhabditis elegans. AB - Dystrophin, the protein disrupted in Duchenne muscular dystrophy, forms a transmembrane complex with dystrophin-associated proteins. Dystrobrevins, proteins showing homology to the C-terminal end of dystrophin, and whose function is unknown, are part of the dystrophin complex. We report here that, in the nematode Caenorhabditis elegans, animals carrying mutations in either the dystrophin-like gene dys-1 or the dystrobrevin-like gene dyb-1 display similar behavioral and pharmacological phenotypes consistent with an alteration of cholinergic signalling. These findings suggest that: (1) dystrobrevin and dystrophin are functionally related and (2) their disruption impairs cholinergic signalling. PMID- 10369885 TI - The drastic reduction of SMN protein in SMA I spinal cord motor neurons is not due to inefficient transcription. AB - Spinal muscular atrophy (SMA) is caused by homozygous absence of the telomeric copy of the survival motor neuron (SMNt) gene. SMNt and its homologous centromeric copy (SMNc) encode the SMN protein, which is markedly reduced in SMA I patients. We have performed SMN transcript and protein studies on spinal cord sections of an SMA I patient using in situ hybridization and immunofluorescence. While the amount of protein was negligible, the level of transcripts was comparable with that of controls. These findings suggest that the reduced protein level is not caused by a deficient transcription of the SMNc gene. PMID- 10369886 TI - Allelic expression of the NF2 gene in neurofibromatosis 2 and schwannomatosis. AB - Neurofibromatosis type 2 (NF2) is a genetic disorder characterized by formation of multiple schwannomas and meningiomas due to inactivating mutations in the NF2 tumor suppressor gene on chromosome 22. We describe a polymorphism in the 3' untranslated region of the NF2 gene that is informative in about one-third of individuals. This polymorphism permitted an assessment of the relative expression of NF2 transcripts in lymphoblastoid cell RNA from 22 unrelated NF2 patients heterozygous for a germline NF2 mutation, along with 6 schwannomatosis patients, and 14 unaffected controls. Unequal allelic expression (1.8- to 20-fold) was detected in 15 of the NF2 cases, but in none of the schwannomatosis or control individuals. Underexpression of the NF2 mutant allele was documented for all 6 nonsense or frameshift mutations, 3 of 6 splice mutations, and 1 of 4 missense mutations, which, unexpectedly, was shown to alter the NF2 transcript and create a premature stop codon. In contrast, equal expression or slight overexpression of NF2 mutant alleles was observed for 2 in-frame deletions, 2 splice alterations, and 3 missense mutations. In the remaining 5 cases, the allele representing the mutant transcript was not known. Thus, truncating NF2 mutations, which are the most frequent alterations in NF2 patients and NF2-associated tumors, were associated with underexpression of the mutant allele, whereas the less common in frame alterations usually showed normal or slight overexpression of the mutant transcript. PMID- 10369887 TI - Is the LDL receptor-related protein involved in Alzheimer's disease? AB - LDL-related protein receptor (LRP) is the main receptor in the brain for apolipoprotein E. Moreover, the LRP gene is located on chromosome 12, the site of a potential Alzheimer's disease (AD) locus. We explored the association between the LRP gene and AD in 600 French Caucasian patients (37.8% men, mean age 72.0+/ 8.0 years, mean age at onset 68.7+/-8.1 years) and age-matched controls (n=646, 37.0% men, mean age 72.5+/-8.2 years) and observed an association between a (TTTC) repeat at the 3' end of an Alu sequence in the LRP gene and the risk of developing AD. Three alleles were detected in this population [corresponding to 83, 87, and 91 base pairs (bp)], the 91-bp allele being associated with an increased risk of developing AD [all patients: odds ratio (OR) 1.6, P<0.01; late onset AD: OR 1.8, P<0.01]. This suggests an association between the LRP locus and AD. However, in the light of studies related to the exon 3 LRP polymorphism and given the low strength of the association reported here, a biologically active variant may exist on chromosome 12, either in the LRP gene itself or in another gene in the vicinity. PMID- 10369888 TI - Electronic identification and chromosomal assignment by radiation hybrid mapping of human expressed sequence tags corresponding to new potassium channel genes. AB - Human homologues of 36 Caenorhabditis elegans potassium channels were identified by expressed sequence tag (EST) database searching. This approach was combined with radiation hybrid mapping to localize new potassium channel genes in the human genome. In addition, several ESTs whose location was already known were also identified as cDNAs encoding additional potassium channels. The identification and mapping of all these genes will make them useful tools for mutation detection in neurological as well as other human diseases. PMID- 10369889 TI - Two novel point mutations of mitochondrial tRNA genes in histologically confirmed Parkinson disease. AB - Mutations in mitochondrially encoded tRNA genes have been described in a variety of neurological disorders. One such mutation, the A to G transition at nucleotide position 4336 of the mitochondrial tRNA(Gln) gene, has been associated with both Alzheimer and Parkinson disease. We have now performed a complete sequence analysis of all 22 mitochondrially encoded tRNA genes in 20 cases of histologically proven idiopathic Parkinson disease. Genomic DNA extracted from the substantia nigra of frozen or formalin-fixed and paraffin-embedded brains was used for amplification by polymerase chain reaction followed by automated sequencing. Two new homoplasmic point mutations were detected in the genes for tRNA(Thr) (15950 G/A) and tRNA(Pro) (15965 T/C) in 1 patient each. Restriction enzyme digestion revealed absence of the 15950 G/A mutation in 96 controls and in 40 cases of neuropathologically confirmed Alzheimer disease. The 15965 T/C mutation was shown to be absent from 100 control subjects and 47 Alzheimer cases. In addition to the two novel mutations, six known sequence variants were detected in a total of 6 different patients in the genes for tRNA(Asp) (G7521A, 1), tRNA(Arg) (T10463C, 1), tRNA(LeuCUN) (A12308G, 2), and tRNA(Thr) (A15924G, 1; G15928A, 2), including 1 patient carrying the tRNA(Gln) (A4336G) mutation. The G15950A transition affects position 70 of the aminoacyl acceptor stem of tRNA(Thr), which has been implicated as a recognition element for threonyl-tRNA synthetase and, at least in some tRNAs, in the processing of primary mitochondrial transcripts. The T15965C point mutation in the mitochondrial tRNA(Pro) gene alters position 64 of the TpsiC stem. The corresponding nucleotide in bacterial aminoacyl-tRNAs is involved in the interaction with elongation factor Tu. Thus, the two novel mutations are likely to be of functional relevance and could contribute to dopaminergic nerve cell death in affected individuals. PMID- 10369890 TI - 5-HTTLPR variants not associated with autistic spectrum disorders. AB - To determine whether there is an association of polymorphic variants of the serotonin transporter (5-HTT) gene-linked polymorphic region (5-HTTLPR) and autistic spectrum disorders, we analyzed the 5-HTTLPR genotypes of 72 autistic subjects, 11 fragile X syndrome patients with autistic behavior, 43 normal subjects, and 49 fragile X syndrome non-autistic subjects. The distribution frequency of 5-HTTLPR long allele (L) and the short allele (S) variants showed no differences between subjects. Our findings do not support the hypothesis that polymorphic 5-HTTLPR variants are a susceptibility factor for autistic disorders. PMID- 10369891 TI - Biochemical and phylogenetic analyses of psychrophilic isolates belonging to the Arthrobacter subgroup and description of Arthrobacter psychrolactophilus, sp. nov. AB - During our work on psychrophilic microorganisms we obtained a large collection of new isolates. In order to identify six of these, we examined their growth properties, cell wall compositions, and their 16S rRNA gene sequences. The results showed that all of the isolates are gram-positive, aerobic, contain lysine in their cell walls, and belong to the high mol% G+C Arthrobacter subgroup. Phylogenetic analysis of the 16S rRNA genes grouped five isolates obtained from a small geographical region into a monophyletic clade. Isolate B7 had a 16S rRNA sequence that was 94.3% similar to that of Arthrobacter polychromogenes and 94.4% similar to that of Arthrobacter oxydans. Primary characteristics that distinguish isolate B7 from the Arthrobacter type strain (Arthrobacter globiformis) and A. polychromogenes include lack of growth at 37 degrees C, growth at 0-5 degrees C, the ability to use lactose as a sole carbon source, and the absence of blue pigments. Because of these differences, isolate B7 was chosen as a type strain representing a new Arthrobacter species, Arthrobacter psychrolactophilus. The sixth isolate, LV7, differed from the other five because it did not have the rod/ coccus morphological cycle and was most closely related to Arthrobacter agilis. PMID- 10369892 TI - Heterologous expression of soluble methane monooxygenase genes in methanotrophs containing only particulate methane monooxygenase. AB - The methanotrophs Methylococcus capsulatus (Bath) and Methylosinus trichosporium OB3b contain particulate methane monooxygenase (pMMO) and soluble methane monooxygenase (sMMO) genes. Other methanotrophs such as Methylomicrobium album BG8 and Methylocystis parvus OBBP contain only pMMO genes. Although molecular genetic techniques are poorly developed in methanotrophs, sMMO genes were expressed in methanotrophs normally containing only pMMO genes. This was achieved by conjugation using broad-host-range plasmids containing the native promoter and sMMO genes from Mc. capsulatus (Bath) and Ms. trichosporium OB3b. sMMO genes derived from Ms. trichosporium OB3b were expressed in an active form in Mcy. parvus OBBP and in Mm. album BG8. Therefore, all of the genes required for active sMMO synthesis were contained on the broad-host-range plasmids and were expressed in the heterologous hosts. Constitutive synthesis of pMMO was observed in Mm. album BG8 when grown at high and low copper-to-biomass ratios, while transcription of the recombinant sMMO genes was only observed under growth conditions of low copper-to-biomass ratios. Therefore, the regulatory protein(s) for sMMO synthesis was also present on the plasmid used, or the heterologous host contained a regulatory system for sMMO. Expression of sMMO genes in methanotrophs containing only pMMO will assist further investigations on the expression and regulation of MMO genes in methanotrophs. PMID- 10369893 TI - Metabolic state of Zymomonas mobilis in glucose-, fructose-, and xylose-fed continuous cultures as analysed by 13C- and 31P-NMR spectroscopy. AB - The reasons for the well-known significantly different behaviour of the anaerobic, gram-negative, ethanologenic bacterium Zymomonas mobilis during growth on fructose (i.e. decreased growth and ethanol yields, increased by-product formation) as compared to that on its second natural substrate, glucose, have remained unexplained. A xylose-fermenting recombinant strain of Z. mobilis that was recently constructed in our laboratory also unexpectedly displayed an increased formation of by-products and a strongly reduced growth rate as compared to the parent strain. Therefore, a comprehensive study employing recently developed NMR-based methods for the in vivo analysis of intracellular phosphorylated pool sizes and metabolic fluxes was undertaken to enable a global characterization of the intracellular metabolic state of Z. mobilis during growth on 13C-labelled glucose, fructose and xylose in defined continuous cultures. The 13C-NMR flux analysis indicated that ribose 5-phosphate is synthesized via the nonoxidative pentose phosphate pathway in Z. mobilis, and it identified a metabolic bottleneck in the recombinant xylose-fermenting Z. mobilis strain at the level of heterologous xylulokinase. The 31P-NMR analyses revealed a global alteration of the levels of intracellular phosphorylated metabolites during growth on fructose as compared to that on glucose. The results suggest that this is primarily caused by an elevated concentration of intracellular fructose 6 phosphate. PMID- 10369894 TI - The acuH gene of Aspergillus nidulans, required for growth on acetate and long chain fatty acids, encodes a putative homologue of the mammalian carnitine/acylcarnitine carrier. AB - The Aspergillus nidulans acuH gene, required for growth on acetate and long-chain fatty acids, was cloned by complementation of the acuH13 mutation. Northern blotting analysis showed that transcription of the acuH gene occurs in acetate grown mycelium and at higher levels in oleate-grown mycelium, but not during growth on glucose minimal medium. The acuH gene encodes a protein of 326 amino acids that belongs to the mitochondrial carrier family. The ACUH protein contains three related segments of approximately 100 amino acids in length, each segment comprising two hydrophobic domains that are probably folded into two transmembrane alpha-helices linked by an extensive polar region. Sequence comparisons suggest that the acuH gene of A. nidulans encodes the homologue of the carnitine/acylcarnitine carrier of rat and man. The uncharacterised proteins YOR100C of Saccharomyces cerevisiae, COLT of Drosophila melanogaster, and DIF-1 of Caenorhabditis elegans also seem to be homologues of ACUH. In addition to the motifs present in all members of the mitochondrial carrier family, we propose the highly conserved motif R(A,S)(V,F)PANAA(T,C)F within the sixth hydrophobic domain of these proteins as the characteristic feature of the carnitine carrier subfamily. The proposed function of the ACUH protein is the transport of acetylcarnitine molecules from the cytosol to the mitochondrial matrix, a process required during growth on acetate or on long-chain fatty acids. PMID- 10369895 TI - Lapstatin, a new aminopeptidase inhibitor produced by Streptomyces rimosus, inhibits autogenous aminopeptidases. AB - Lapstatin, a low-molecular-weight aminopeptidase inhibitor, was purified to homogeneity from Streptomyces rimosus culture filtrates. The purification procedure included extraction with methanol, followed by chromatography on Dowex 50WX4, AG50WX4, and HPLC RP C18 columns. By amino acid analysis, mass spectrometry, and NMR spectroscopy, the structure of lapstatin was shown to be 3 amino-2-hydroxy-4-methylpentanoylvaline. Lapstatin inhibited the extracellular leucine aminopeptidases from Streptomyces rimosus, Streptomyces griseus, and Aeromonas proteolytica with an IC50 in the range of 0.3-2.4 microM. IC50 values for other enzymes tested were at least tenfold higher. Leucine aminopeptidase from Streptomyces griseus was inhibited in a competitive manner, with an inhibition constant of 5 x 10(-7) M. Lapstatin is the first low-molecular-weight compound isolated from streptomycetes shown to inhibit an autogenous aminopeptidase. PMID- 10369896 TI - Selective enrichment and characterization of Roseospirillum parvum, gen. nov. and sp. nov., a new purple nonsulfur bacterium with unusual light absorption properties. AB - A new type of phototrophic purple bacterium, strain 930I, was isolated from a microbial mat covering intertidal sandy sediments of Great Sippewissett Salt Marsh (Woods Hole, Mass., USA). The bacterium could only be enriched at a wavelength of 932 (+/- 10) nm. Cells were vibrioid- to spirilloid-shaped and motile by means of bipolar monotrichous flagellation. The intracytoplasmic membranes were of the lamellar type. Photosynthetic pigments comprised bacteriochlorophyll a and the carotenoids spirilloxanthin and lycopenal. The isolated strain exhibited an unusual, long-wavelength absorption maximum at 911 nm. Sulfide or thiosulfate served as electron donor for anoxygenic phototrophic growth. During growth on sulfide, elemental sulfur globules formed outside the cells. Elemental sulfur could not be further oxidized to sulfate. In the presence of sulfide plus bicarbonate, fructose, acetate, propionate, butyrate, valerate, 2 oxoglutarate, pyruvate, lactate, malate, succinate, fumarate, malonate, casamino acids, yeast extract, L(+)-alanine, and L(+)-glutamate were assimilated. Sulfide, thiosulfate, or elemental sulfur served as a reduced sulfur source for photosynthetic growth. Maximum growth rates were obtained at pH 7.9, 30 degrees C, 50 micromol quanta m-2 s-1 of daylight fluorescent tubes, and a salinity of 1 2% NaCl. The strain grew microaerophilically in the dark at a partial pressure of 1 kPa O2. The DNA base composition was 71.2 mol% G + C. Sequence comparison of 16S rRNA genes indicated that the isolate is a member of the alpha-Proteobacteria and is most closely related to Rhodobium orientis at a similarity level of 93.5%. Because of the large phylogenetic distance to known phototrophic species of the alpha-Proteobacteria and of its unique absorption spectrum, strain 930I is described as a new genus and species, Roseospirillum parvum gen. nov. and sp. nov. PMID- 10369897 TI - Morpholine-induced formation of L-alanine dehydrogenase activity in Mycobacterium strain HE5. AB - An NAD-dependent, morpholine-stimulated L-alanine dehydrogenase activity was detected in crude extracts from morpholine-, pyrrolidine-, and piperidine-grown cells of Mycobacterium strain HE5. Addition of morpholine to the assay mixture resulted in an up to 4. 6-fold increase of L-alanine dehydrogenase activity when L-alanine was supplied at suboptimal concentration. L-alanine dehydrogenase was purified to near homogeneity using a four-step purification procedure. The native enzyme had a molecular mass of 160 kDa and contained one type of subunit with a molecular mass of 41 kDa, indicating a tetrameric structure. The sequence of 30 N terminal amino acids was determined and showed a similarity of up to 81% to that of various alanine dehydrogenases. The pH optimum for the oxidative deamination of L-alanine, the only amino acid converted by the enzyme, was determined to be pH 10.1, and apparent Km values for L-alanine and NAD were 1.0 and 0.2 mM, respectively. Km values of 0. 6, 0.02, and 72 mM for pyruvate, NADH, and NH4+, respectively, were estimated at pH 8.7 for the reductive amination reaction. PMID- 10369899 TI - Linear alkanesulfonates as carbon and energy sources for gram-positive and gram negative bacteria. AB - Several bacteria from soil and rainwater samples were enriched and isolated with propanesulfonate or butanesulfonate as sole carbon and energy source. Most of the strains isolated utilized nonsubstituted alkanesulfonates with a chain length of C3-C6 and the substituted sulfonates taurine and isethionate as carbon and energy source. A gram-positive isolate, P40, and a gram-negative isolate, P53, were characterized in more detail. Phylogenetic analysis grouped strain P40 within group IV of the genus Rhodococcus and showed a close relationship with Rhodococcus opacus. After phylogenetic and physiological analyses, strain P53 was identified as Comamonas acidovorans. Both bacteria also utilized a wide range of sulfonates as sulfur source. Strain P40, but not strain P53, released sulfite into the medium during dissimilation of sulfonated compounds. Cell-free extracts of strain P53 exhibited high sulfite oxidase activity [2.34 U (mg protein)-1] when assayed with ferricyanide, but not with cytochrome c. Experiments with whole cell suspensions of both strains showed that the ability to dissimilate 1 propanesulfonate was specifically induced during growth on this substrate and was not present in cells grown on propanol, isethionate or taurine. Whole-cell suspensions of both strains accumulated acetone when oxidizing the non-growth substrate 2-propanesulfonate. Strain P40 cells also accumulated sulfite under these conditions. Stoichiometric measurements with 2-propanesulfonate as substrate in oxygen electrode experiments indicate that the nonsubstituted alkanesulfonates were degraded by a monooxygenase. When strain P53 grew with nonsubstituted alkanesulfonates as carbon and energy source, cells expressed high amounts of yellow pigments, supporting the proposition that an oxygenase containing iron sulfur centres or flavins was involved in their degradation. PMID- 10369898 TI - Generation and initial characterization of Pseudomonas stutzeri KC mutants with impaired ability to degrade carbon tetrachloride. AB - Under iron-limiting conditions, Pseudomonas stutzeri KC secretes a small but as yet unidentified factor that transforms carbon tetrachloride (CT) to CO2 and nonvolatile products when activated by reduction at cell membranes. Pseudomonas fluorescens and other cell types activate the factor. Triparental mating was used to generate kanamycin-resistant lux::Tn5 recombinants of strain KC. Recombinants were streaked onto the surface of agar medium plugs in microtiter plates and were then screened for carbon tetrachloride degradation by exposing the plates to gaseous 14C-carbon tetrachloride. CT+ recombinants generated nonvolatile 14C labeled products, but four CT- recombinants did not generate significant nonvolatile 14C-labeled products and had lost the ability to degrade carbon tetrachloride. When colonies of P. fluorescens were grown next to colonies of CT+ recombinants and were exposed to gaseous 14C-carbon tetrachloride, 14C-labeled products accumulated around the P. fluorescens colonies, indicating that the factor secreted by CT+ colonies had diffused through the agar and become activated. When P. fluorescens was grown next to CT- colonies, little carbon tetrachloride transformation was observed, indicating a lack of active factor. Expression of lux reporter genes in three of the CT- mutants was regulated by added iron and was induced under the same iron-limiting conditions that induce carbon tetrachloride transformation in the wild-type. PMID- 10369900 TI - Analysis of the expression and function of the sigmaB-dependent general stress regulon of Bacillus subtilis during slow growth. AB - Glucose-limited continuous cultures were used to analyze sigmaB activity at decreasing growth rates. Expression of the sigmaB-dependent genes gsiB and ctc started to increase at a growth rate of 0.2 h-1, and both genes were induced approximately fivefold at a growth rate of 0.1 h-1 as compared to expression at the maximal growth rate. However, maximal sigmaB activity was only reached when the growth stopped as a result of the exhaustion of the carbon and energy source glucose. During glucose-limited growth, increased expression of the general stress regulon at growth rates below 0.2 h-1 did not provide wild-type cells with a growth advantage over sigB mutants. Instead, expression of the stress regulon seems to constitute a significant burden during glucose-limited growth, resulting in a selective growth advantage of the sigB mutant as compared to the wild-type at a growth rate of 0.08 h-1. PMID- 10369901 TI - Diffuse neonatal hemangiomatosis with extensive involvement of the brain and cervical spinal cord. AB - BACKGROUND: Diffuse neonatal hemangiomatosis (DNH) is a rare disorder first recognized at birth or during the neonatal period. DNH is characterized by numerous cutaneous and visceral hemangiomas involving three or more organ systems. MATERIALS AND METHODS: Although the skin and liver are most frequently affected, we present a case of DNH demonstrating an unusual predilection for the central nervous system (CNS). RESULTS AND CONCLUSION: We report the imaging findings in a patient with this disorder, paying particular attention to the features seen on cranial sonography and spinal MR imaging. PMID- 10369902 TI - Application of a three-point method for water-fat MR imaging in children. AB - BACKGROUND: Effective fat suppression is desirable in clinical magnetic resonance imaging. Conventional frequency selective fat suppression is dependent on accurate prescan shimming and is subject to artifacts due to magnetic field inhomogeneity. Quadrature three-point water-fat imaging with direct phase encoding is an alternative technique for fat suppression that has been previously described in adult volunteers and patients. OBJECTIVE: To evaluate the use of three-point water-fat imaging with direct phase encoding for fat-suppressed MR scans in children. MATERIALS AND METHODS: Sixty-two three-point water-fat imaging studies were performed in 55 children 2 months to 18 years old. T 1-weighted fat suppressed (water) images from this sequence were compared with frequency selective fat-suppressed images obtained in 15 patients. The reliability and subjective quality of the sequence were assessed in the remaining 47 cases. RESULTS: High-quality fat suppression was achieved in all anatomic sites studied, even where frequency selective fat-suppression failed due to magnetic susceptibility artifact. The three-point water-fat sequence was visually preferred to the frequency selective fat saturation technique in 15/15 cases. CONCLUSION: Three-point water-fat imaging has replaced the conventional frequency selective technique for fat suppression on T 1-weighted MR imaging at our institution. PMID- 10369903 TI - Reversibility of hyperintense globus pallidus on T 1-weighted MRI follow- ing surgery for a portosystemic shunt in an 8-year-old girl. AB - An 8-year-old Japanese girl with a portosystemic shunt had shown hyperammonaemia since she was 3 years of age. MRI of her brain showed bilateral hyperintense globus pallidus. A portosystemic shunt was evident on US and angiography. She underwent surgical banding of the shunt, after which the lesion and clinical symptoms disappeared. PMID- 10369904 TI - Direct coronal CT scanning of the neonatal chest. AB - BACKGROUND: The purpose of this study was to attempt to improve upon conventional coronal computed tomographic (CT) images of neonatal chest which to date have been made by reformatting thinly spaced axial images. MATERIALS AND METHODS: Nine neonates were studied by direct coronal CT scans with the patients' long axis 90 degrees to the scanning table. They were studied to further define their thoracic abnormalities detected on plain film. Spiral CT and cine scan (Imatron) were utilized. RESULTS: Congenital lung lesions such as congenital cystic adenomatoid malformation could not be diagnosed but their anatomical location could be accurately depicted, enabling easier surgical planning. The arterial supply to bronchopulmonary sequestrations was also identifiable. Tracheobronchial abnormalities such as tracheobronchus and bronchial atresia were also identifiable. Causing of air trapping, both intrinsic such as an atretic bronchus and extrinsic such as vascular compression were readily demonstrated. CONCLUSION: With neither special devices nor paraphernalia, the described method of direct coronal CT scans were both feasible and provided significant information. This technique allows for improved assessment of the tracheobronchial tree and more accurate detection, localization, and characteristics of lesions adjacent to the diaphragm. PMID- 10369905 TI - Familial pulmonary hypoplasia: plain film appearances with histopathological correlation. AB - Familial pulmonary hypoplasia is a rare cause of bilateral pulmonary hypoplasia. We describe the plain film appearances and correlate these with the histopathological findings in an infant who survived 5 weeks. In this case, improved technology prolonged survival, allowing the radiological appearance of the primitive lung architecture to become more clearly defined. PMID- 10369906 TI - Inter- and intra-observer variability in the assessment of atelectasis and consolidation in neonatal chest radiographs. AB - BACKGROUND: Radiology is an essential part of neonatal intensive care. Interpretation of chest radiographs frequently contributes to respiratory management of neonates, but there has been little assessment of the consistency of this interpretation. OBJECTIVE: To assess the inter- and intra-observer variability for the reporting of atelectasis and/or consolidation in neonatal chest radiographs. MATERIALS AND METHODS: A total of 585 chest radiographs from the 220 babies ventilated in our nursery over a 2-year period were coded by two radiologists for generalised, lobar and segmental atelectasis and/or consolidation. Two months later one of the radiologists re-coded a random sample of these films (n = 117, 20 %). Agreement was assessed by the kappa statistic and by proportions of agreement for normality and abnormality. RESULTS: The reported incidence of focal atelectasis was low (5-6 %). Focal changes of any nature were found in 21-26 % of films. Inter-observer agreement was fair to moderate (kappa = 0.25-0.44). Intra-observer agreement was mostly moderate to good (kappa = 0.38 0.66). CONCLUSION: The poor inter-observer agreement for the diagnosis of pulmonary parenchymal abnormalities on chest radiographs of neonates receiving intensive care suggests that abnormalities should be described rather than diagnoses given or that a list of differential diagnoses be offered. When research involves radiographic interpretation, the potential lack of consistency in reporting abnormalities must be borne in mind. PMID- 10369907 TI - MR cholangiography in children with autosomal recessive polycystic kidney disease. AB - BACKGROUND: Magnetic resonance cholangiography (MRC) is a relatively new, non invasive imaging technique of the biliary tree that has shown good correlation with endoscopic retrograde cholangiopancreatography. The liver manifestation of autosomal recessive polycystic kidney disease (ARPKD) is congenital hepatic fibrosis (CHF). CHF may be accompanied by Caroli's disease, which is characterised by a non-obstructive dilation of the intrahepatic bile ducts. OBJECTIVE: A prospective study was conducted to determine the presence and extent of Caroli's disease in children with ARPKD. MATERIALS AND METHODS: Seven children with ARPKD aged from 3.0 to 10. 1 years were examined. CHF was confirmed in all biopsied cases (5 of 7). All children had been followed by repeated abdominal US examinations for many years. The MR examination included a morphological imaging study using a T2-weighted turbo spin-echo sequence and a heavily T2-weighted inversion-recovery turbo spin-echo sequence with three-dimensional maximum intensity projection (MIP) reconstructions for MRC. RESULTS: The diagnosis of Caroli's disease could be made in one case by US; in two other children Caroli's disease was suspected, but the differentiation from hepatic cysts was not possible. By MRC, Caroli's disease could be diagnosed in three of seven children. Furthermore, MRC with MIP reconstructions demonstrated the extent of the disease by showing the entire biliary tree from different angles. CONCLUSIONS: MRC is a valuable method to establish the diagnosis and demonstrate the extent of Caroli's disease. PMID- 10369908 TI - The duodenal dimple: a specific fluoroscopic correlate to the duodenal web. AB - PURPOSE: We describe a new fluoroscopic sign to aid in the diagnosis of an obstructing duodenal web and its attachment site. MATERIALS AND METHODS: During an upper GI series of a neonate, a nasogastric tube was passed into an obstructed duodenum and barium injected. The tube, pressing on an obstructing web, caused dimpling of the duodenal contour at the attachment point of the web to the duodenal wall. The same maneuver at surgery caused identical dimpling. DISCUSSION: While the maneuver is described in surgical textbooks, there has been no radiologic correlate. The "duodenal dimple" is a new fluoroscopic sign of a duodenal web and its attachment point to the duodenal wall. PMID- 10369909 TI - Ultrasonographic assessment of Salmonella enterocolitis in children. AB - BACKGROUND: Salmonella enterocolitis (SE) is one of the important causes of acute infectious diarrhoea. Imaging studies are rarely performed on these patients. Consequently, ultrasound (US) features of SE are controversial. OBJECTIVE: To identify the clinical significance of US in the evaluation of SE. MATERIAL AND METHODS: Abdominal US was performed in 15 patients with SE and 9 patients with Rotavirus enterocolitis (RE). RESULTS: Ascites was present in 60 % and mural thickening of the colon in 40 % of patients with SE on abdominal US, whereas we could not identify these features in patients with RE. In patients with SE, colonic wall thickening; and ascites, the levels of C-reactive protein (CRP) were significantly higher as compared to patients with SE and no colonic wall thickening or ascites. Also, the stool occult blood test was positive more often in patients with colonic wall thickening and ascites on US than in patients without these findings. The colonic wall thickness significantly correlated with CRP and stool occult blood level. CONCLUSIONS: US is able to identify pathological changes in bowel and intra-abdominal spaces. The US findings of ascites and colonic wall thickening may be useful for determining the severity of SE. PMID- 10369910 TI - Intestinal blood-flow velocity in uncomplicated preterm infants during the early neonatal period. AB - BACKGROUND: Intestinal blood-flow changes after birth. Objective. To elucidate the factors influencing intestinal blood-flow velocity in preterm infants during the early neonatal period. MATERIALS AND METHODS: We measured blood-flow velocity in the superior mesenteric artery by pulsed Doppler US in 44 uncomplicated infants with a gestational age of less than 34 weeks and from 1 to 6 days of age. RESULTS: Time-averaged mean blood-flow velocity significantly increased with age from 1 to 6 days old. There was a significant correlation of time-averaged mean blood-flow velocity with birth weight at 1, 2, 4, 5 and 6 days of age and with the amount of enteral feeding from 4 to 6 days of age. Multivariate analysis showed that partial correlation of time-averaged mean blood-flow velocity with birth weight at 2 days of age and that with the amount of enteral feeding at 5 days of age were significant. End-diastolic blood-flow velocity was significantly lower at 1 day of age in infants with patent ductus arteriosus than those without it. CONCLUSIONS: Age, birth weight, the amount of enteral feeding and patent ductus arteriosus are included in the determinants of intestinal blood-flow velocity in preterm infants. PMID- 10369911 TI - Punctate epiphyses associated with Turner syndrome. AB - The radiographic observation of stippled calcification in cartilage defines the chondrodysplasia punctata group of bone dysplasias. Several other diseases may be associated with the radiographic finding of punctate epiphyses, usually uncommonly - for example, trisomy 21. Other more subtle chromosomal abnormalities also associated with punctate epiphyses include microdeletions of the X chromosome. A case of Turner syndrome with punctate calcification of the epiphyses is described. PMID- 10369912 TI - A simple bone cyst containing secretory cells in its lining membrane in a patient with polyostotic fibrous dysplasia. AB - An 11-year-old boy, severely affected with polyostotic fibrous dysplasia, showed radiographically rapid expansion of a cystic lesion in his right humerus. At biopsy, there was an extraordinarily thin shell of bone and a cavity encapsulated by a hypertrophic fibrous membrane and filled with yellow serous fluid. Histologically, in addition to typical features of fibrous dysplasia, the fibrous capsule membrane was composed of proliferated mesenchymal cells characteristic of the affected bone. Ultrastructurally, many secretory granules were observed in numerous cytoplasmic vacuoles in the capsular cells as well as in the cultured cells isolated from the evacuated fluid. PMID- 10369913 TI - Reversible ureteric and pelvicalyceal calcification in a 12-year-old boy. AB - A 12-year-old boy was shown on CT to have unexplained pelvicalyceal and ureteric calcification which resolved over a 16-month period. PMID- 10369914 TI - Eosinophilic cystitis following an infected urachal remnant. PMID- 10369915 TI - Decompression of pneumatocele in a neonate by percutaneous catheter placement. PMID- 10369916 TI - The human CD38 gene: polymorphism, CpG island, and linkage to the CD157 (BST-1) gene. AB - CD38 is a leukocyte activation antigen and ectoenzyme [NAD(P)+ glycohydrolase; EC 3.2.2.6] involved in numerous immune functions. The human CD38 gene is complex [eight exons, >80 kilobases (kb) long] located on Chromosome 4p15, and part of the eukaryotic NAD+ glycohydrolase/ADP-ribosyl cyclase gene family. Because of the increasing relevance of the CD38 molecule in the host immune response to infectious, tumoral, and metabolic diseases, we investigated the genetic variability and linkage of the human CD38 locus. We report that (1) the restriction endonuclease Pvu II identifies a bi-allelic polymorphism here defined as formed by the alleles CD38*A (12 kb) and CD38*B (9/2.5 kb); (2) their frequency in the healthy Italian Caucasian population is 14% and 86%, respectively; (3) the polymorphic Pvu II site is located at the 5' end of the first intron of the CD38 gene; (4) in conjunction with the polymorphic site, we identified a 900 base pair CpG island associated with the CD38 gene, with two potential Sp1 binding sites; (5) the CpG island may play a role in the regulation of CD38 expression and is hypomethylated in various cell lines; (6) by pulsed field gel electrophoresis we show that CD38 and its paralogue, the bone-marrow stromal cell antigen BST-1 (CD157), map to the same 800 kb Avi II fragment, indicating that the two human ecto-NADase genes are closely linked. PMID- 10369917 TI - Population study of allelic diversity in the human MHC class I-related MIC-A gene. AB - The polymorphism of major histocompatibility complex (MHC) class I HLA-A, -B, and -C molecules may have evolved through pathogen-driven selection of alleles with diverse peptide-binding specificities. Two MHC-encoded molecules that are distantly related to class I, MIC-A and MIC-B, do not function in the presentation of pathogen-derived peptides to T cells with alphabeta T-cell receptors (TCRs), but are broadly recognized by intraepithelial T cells with gammadelta TCRs. However, both MIC-A and MIC-B are polymorphic, displaying an unusual distribution of a number of variant amino acids in their extracellular alpha1, alpha2, and alpha3 domains. In order to further define the polymorphism of MIC-A, we examined its alleles among 275 individuals with common and rare HLA genotypes. Of 16 previously defined alleles, 12 were confirmed and 5 new alleles were identified. A two-by-two analysis of MIC-A and HLA-B alleles uncovered a number of statistically significant associations. These results confirm and extend previous knowledge on the polymorphism of MIC-A. The strong positive linkage of certain MIC-A and HLA-B alleles may have implications for studies related to MHC-associated diseases and transplantation. PMID- 10369918 TI - MIC-A allele profiles and HLA class I associations in Behcet's disease. AB - Recently a new family of non-classical MHC molecules, the MHC class I chain related protein (MIC), encoded by genes located in the major histocompatability complex have been identified. On the basis of the location of MIC genes and the structure and expression of MIC molecules it has been postulated that MIC may be a susceptibility factor in Behcet's disease (BD). We investigated the association of the 16 described external domain alleles and the transmembrane triplet repeats of MIC-A with BD in a Middle Eastern population. DNA from ninety-five patients and 102 age- and sex-matched controls were analyzed by polymerase chain reaction using allele specific primers. Our results show an increase of MIC-A*009 in the BD patient group 44/95 (46%) compared with controls 24/102 (24%) (chi2=11.3, OR=2.8, P=0.00078). MIC-A*009 was also found to be strongly associated with HLA B51 in the patients 39/44 (88%) when compared with controls 10/24 (42%) (chi2=4, P=0.04). MIC-A*009 was also found in linkage disequilibrium with HLA-B52, but only in controls. The A6 form of a MIC-A transmembrane triplet repeat was found to be significantly raised in the patients (80/95; 84%;) compared with controls (58/102, 57%) (chi2=17.5, OR=4, P=0.000028). Although the MIC-A associations described are highly significant, the association with HLA-B51 independently remains the most significant factor (chi2=56.8, P<10(-6)). The data suggests that as both MIC-A*009 and A6 are in strong linkage disequilibrium with HLA-B51, they are unlikely to be the susceptibility gene for BD but may be markers for additional risk factors. PMID- 10369919 TI - Characterization of a polymorphism in the coding region of the human C5a anaphylatoxin receptor. AB - A polymorphism was identified in the coding region of the human C5a anaphylatoxin receptor gene leading to C to T transition at nucleotide position 450 (a silent substitution in the Ala150 codon, GCC to GCT). Its distribution was studied in a population of healthy volunteers from the Quebec city region (prevalence of 2.8%) and among patients with end-stage renal failure who had previously undergone renal graft (prevalence 1.4%, not significantly different from that of the control group). This new marker provides a valuable tool to assess the risk for putative C5a-associated disorders with genetic determinism. PMID- 10369920 TI - MIC-A polymorphism in Japanese and a MIC-A-MIC-B null haplotype. AB - A polymorphic gene, MIC-A, is one of the MIC family of genes which is composed of a group of homologous genes interspersed in the class III and class I regions of the major histocompatibility complex. MIC-A is located 46 kilobases (kb) centromeric of HLA-B, and is preferentially expressed in the epithelial cells and intestinal mucosa. Recently, MIC-A and the closely related MIC-B were reported as the molecules that conferred specificity in the recognition by the Vdelta1gammadeltaT cells. In the present study, polymorphic exons 2, 3, and 4 of the MIC-A gene were analyzed using the polymerase chain reaction-single-strand conformation polymorphism method. The number of patterns found in exons 2, 3, and 4 were 5, 6, and 4, respectively, in 114 healthy Japanese subjects. Eight MIC-A alleles were observed in Japanese individuals, among which one, tentatively named MIC-AMW, has not previously been reported. There was a strong linkage disequilibrium between MIC-A and HLA-B loci: each MIC-A allele showed strong association with a particular HLA-B group. In contrast, B*3901 showed association with multiple MIC-A alleles. Furthermore, the existence of a MIC-A-MIC-B null haplotype, which is associated with HLA-B*4801, was identified. In this haplotype, a large-scale deletion (of approximately 100 kb) including the entire MIC-A gene was indicated and the MIC-B gene possessed a stop codon. PMID- 10369921 TI - Cytonuclear disequilibria in wild populations of rabbit (Oryctolagus cuniculus L.) suggest unequal allele turnover rates at the b locus (IGKC1). AB - DNA sequence comparisons suggest that evolutionary rates at the rabbit IGKC1 locus can differ among allelic lineages. Here we address the question of whether population turnover rates can vary among IGKC1 alleles. We studied the distribution of sixteen IGKC1 (or b-locus) allotypes in areas comprising the aboriginal species range (Iberian peninsula). Rabbits in this area belong to one of two distantly related mitochondrial lineages (mtDNA types) A and B. In the more recent distribution area of the species, all rabbits belong to the mtDNA type B lineage, and IGKC1 alleles b4 and b5 comprise over 90% of the gene pool. These two alleles are also predominant in areas of mtDNA type B prevalence within the Iberian range. However, in areas of mtDNA type A prevalence, the b4 and b5 allotypes are rare or absent; they apparently have been replaced by serologically related, but distinct, 'endemic' variants. The cytonuclear disequilibria were highly significant, also within the subsample consisting of populations from Spain. These observations suggest that allelic persistence times for the predominant IGKC1 lineages could be shorter than the divergence time of the major mtDNA lineages A and B. In contrast, the relative gene frequencies of the IGKC1 allele b9 were similar among the type A and type B rabbits; it was present in most populations at low frequency. In consequence, persistence times of the b9 allele appear to be longer than the divergence time of lineages A and B. The data reported here are in agreement with the DNA sequence data, providing further proof that the molecular clock can run at different rates among allelic lineages at the rabbit IGKC1 locus. PMID- 10369922 TI - Genomic organization of the HSET locus and the possible association of HLA-linked genes with immotile cilia syndrome (ICS). AB - The kinesin-related protein (HSET) gene belongs to the kinesin superfamily, the members of which are involved in cellular transport processes. The HSET gene product was previously characterized by partial cDNA sequencing. The gene is located on the short arm of human Chromosome 6 (6p21.3), at the centromeric end of the major histocompatibility complex. Here, we report the genomic structure of the complete HSET gene together with its flanking loci. Sequence analysis of the 40 kilobase (kb) cosmid clone containing the HSET gene also revealed the presence of several new genes not related to the kinesin superfamily. These include a 60S ribosomal protein L35A-like pseudogene (rPL35A-like) on the telomeric side and a polycomb-like gene (PHF1), a copper tolerance-like gene (CUTA1) and the 5' part of the synaptic ras-GTPase-activating protein (SynGAP) gene centromeric of HSET. In addition, a complete 60S ribosomal protein L12-like (rPL12L) gene in intron 3 of the HSET gene was identified which appears to have an open reading frame. The possible involvement of the HSET gene and a beta-tubulin gene (TUBB) in the pathogenesis of immotile cilia syndrome (ICS) was studied by screening two unrelated ICS families with microtubular defects and suspected HLA linkage for mutations within the HSET gene and the TUBB gene. Four single base substitutions were detected in the HSET gene, and none in the TUBB gene. On the basis of these data, a role of the HSET and TUBB products in the pathogenesis of ICS in the two families is unlikely. PMID- 10369923 TI - Selection in favor of the Ped fast haplotype occurs between mid-gestation and birth. AB - The preimplantation embryo development (Ped) gene that encodes the class Ib major histocompatibility complex protein Qa-2 influences the rate of embryonic cleavage during the preimplantation stages of development. Embryos from strains of mice that lack the Ped gene cleave slowly, while embryos that have a functional Ped gene cleave more rapidly. This effect is observed both in vivo and in vitro with the Ped fast haplotype showing dominance over the Ped slow haplotype. The Ped gene is associated with pleiotropic effects on reproduction. Certain strains of mice lacking the Ped gene (Ped slow) have smaller litters and the pups weigh less at birth and at weaning. Previously our laboratory reported that in litters derived from Ped fast/slow F1 mice backcrossed to the slow/slow parent, there were significantly more Ped fast pups than the 50% expected, at two months of age. This implies that there is selection in favor of the Ped fast haplotype at some point during development. The present study was designed to determine at what point during development selection occurs. Using a polymerase chain reaction assay, we determined that selection does not occur by days post coitus 14.5. However, our results show that there are significantly more Ped fast pups than Ped slow pups remaining in backcross litters just after birth, indicating that selection in favor of the Ped fast haplotype occurs between day 14.5 and birth. PMID- 10369924 TI - The central MHC gene IKBL carries a structural polymorphism that is associated with HLA-A3,B7,DR15. AB - Susceptibility to several disorders, including insulin-dependent diabetes mellitus and multiple sclerosis, has been associated with alleles of HLA class II genes and loci in the TNF cluster in the central major histocompatibility complex (MHC) region. As recombination within this region is rare, it is difficult to determine which genes are important. This will be facilitated by the identification of functional polymorphisms. Hence we are sequencing reverse transcription-polymerase chain reaction products derived from central MHC genes in well characterized and conserved ancestral haplotypes. Here we address the IKBL gene, which lies near the TNF cluster at the telomeric end of the central MHC. Although the IKBL cDNA sequence was conserved between most ancestral haplotypes, a synonymous nucleotide substitution, a 3' untranslated region substitution, and a single nonsynonymous substitution were identified. The latter (IKBL+738) was present in multiple examples of the 7.1 haplotype [HLA-A3, B7, DR2 (DR15)] and resulted in a cysteine to arginine substitution in a predicted protein kinase C phosphorylation site. This polymorphism did not occur in 18 other common haplotypes from the 10th International Histocompatibility Workshop and thus appears haplospecific. A role for IKBL+738 in the association between HLA-A3,B7,DR2(DR15) and susceptibility to multiple sclerosis is discussed. PMID- 10369925 TI - The H-mshi antigen is conserved among standard BALB/cBy, C57BL/6J, and wild derived CAST/Ei and SPRET/Ei inbred strains of mice. AB - The recessive male sterility and histoincompatibility mutation (mshi) arose spontaneously in the standard inbred mouse strain BALB/cBy. In addition to generating sterility in homozygous males, mshi controls the loss of a minor histocompatibility antigen designated H-mshi. To determine whether the H-mshi antigen normally expressed by the BALB/cBy strain (H-mshi(c)) is the same as or different from the antigen (H-mshi(x)) expressed by the standard inbred C57BL/6J strain or the wild-derived CAST/Ei and SPRET/Ei strains, animals heterozygous for the mutant antigen-loss allele (H-mshi-) and H-mshi(x) were grafted with tail skin from BALB/cBy mice. The long-term retention of grafts by these hosts indicates that the H-mshi antigen encoded by the BALB/cBy, C57BL/6J, CAST/Ei, and SPRET/Ei strains is histogenically identical. Conservation of this minor histocompatibility antigen among these evolutionarily diverse strains suggests that H-mshi encodes a functionally important cellular product(s). PMID- 10369926 TI - The chicken 9E3/CEF4 CXC chemokine is the avian orthologue of IL8 and maps to chicken chromosome 4 syntenic with genes flanking the mammalian chemokine cluster. AB - The gene encoding the chicken chemokine 9E3/CEF4 was cloned, sequenced, and mapped; 9E3/CEF4 was the first nonmammalian cytokine cDNA to be cloned and has significant amino acid identity with both human IL8 and human GROalpha. These results show that this cytokine is chicken IL8 and not GROalpha. The exon:intron structure of chicken IL8 corresponds almost exactly to that of human IL8 and differs from those of other known mammalian CXC chemokine genes. Analysis of the predicted amino acid sequence suggests that overall protein structure is conserved between human and chicken IL8, but that the receptor binding sites are not. Genetic distance analysis also suggests that this gene encodes chicken IL8. A number of potential regulatory sequences similar to those found in human IL8 have been identified in the promoter. These include (5'-3') a hepatocyte NF-1 binding site, an NF-kappaB binding site, and a TATAAA box. The human AP-1 binding site and CCAT box are poorly conserved in the promoter of the chicken gene, but there are other potential AP-1 binding sites and a potential CCAT box. The human IRF-1 and octamer binding sites seem to be absent. However, the chicken gene promoter contains a GATA motif not present in the promoter of human IL8. Sequence comparisons also identify conserved regions in the promoter that may function as transcription factor binding sites as yet undescribed in the human IL8 promoter. Promoter sequence polymorphisms have been identified in chicken lines C and 61, but neither lie in any of the regulatory regions mentioned above. Chicken IL8 contains nine repeats of the "instability" motif ATTTA in the 3' untranslated region (UTR) in exon 4. A multiple restriction single-stranded conformational polymorphism was identified which enabled chicken IL8 to be genetically mapped to Chromosome (Chr) 4, linked to SPP1 and ALB1, and thus showing conserved synteny with mouse Chr 5 and human Chr 4. This is the first nonmammalian chemokine gene to be genetically mapped. PMID- 10369927 TI - Expression analysis of new Ly49 genes: most transcripts of Ly49j lack the transmembrane domain. AB - Five new Ly49 genes, named Ly49j-n, have recently been identified in C57BL/6 mice. This study examined the expression of three of these new genes, Ly49j, k, and n. To determine whether the Ly49j, k, and n genes were transcribed, gene specific primers were used to amplify cDNA clones for each gene from C57BL/6 interleukin-2-activated natural killer (NK) cell cDNA. A full-length cDNA for Ly49j was detected which encodes a 267 amino acid protein and shares approximately 96% nucleotide identity with Ly49c and i. COS cells transfected with the Ly49j cDNA were shown to react with the monoclonal antibody 8H7, suggesting that the gene likely encodes a functional protein. Many different sized Ly49k and n transcripts were observed, although it is likely that they do not encode functional proteins due to missing exons or severe truncations in the open reading frames. Interestingly, the most abundant Ly49j transcript detected was shown to lack exon 3, which encodes the transmembrane domain. Similar studies performed on the same source of NK cell cDNA using Ly49c- and i-specific primers revealed the presence of transmembrane-less Ly49i transcripts, although at a much lower frequency than observed for Ly49j. We also detected Ly49g and h transcripts lacking the transmembrane domain. Despite the absence of the transmembrane region, the resulting Ly49 transcripts maintain their open reading frames, and therefore could potentially encode cytoplasmic proteins with a role in NK cell function. PMID- 10369928 TI - Opposite orientation of the alpha- and upsilon-chain constant region genes in the immunoglobulin heavy chain locus of the duck. PMID- 10369929 TI - Measurement of dog cytokines by reverse transcription-quantitative competitive polymerase chain reaction. PMID- 10369930 TI - Cis-acting regulation of splenic Art2 gene expression in inbred mouse strains. PMID- 10369931 TI - The gene encoding mouse interleukin-16 consists of seven exons and maps to chromosome 7 D2-D3. PMID- 10369932 TI - Genomic organization and sequence of the H2-T24 gene. PMID- 10369933 TI - CTL and sequence analyses of MHC class IB antigens Qa1(c) (H2-T23(r)) and Qa1(d) (H2-T23(f)). PMID- 10369934 TI - Molecular characterization of cDNAs encoding squirrel monkey (Saimiri sciureus) CD8 alpha and beta chains. PMID- 10369935 TI - MHC characterization of ALR and ALS mice: respective similarities to the NOD and NON strains. PMID- 10369936 TI - Molecular cloning of mouse NKG2A and C. PMID- 10369937 TI - The human Ly-49L gene. PMID- 10369939 TI - A new polymorphism in the 5' flanking region of the human interleukin (IL)-4 gene. PMID- 10369938 TI - Nucleotide sequences of three distinct complementary DNA clones encoding rat class II major histocompatibility complex RT1.D beta-chain proteins. PMID- 10369940 TI - A new microsatellite marker within the promoter region of the MIP-1A gene. PMID- 10369941 TI - Modelling: when and why? PMID- 10369942 TI - Biokinetics and radiation doses for carbon-14 urea in adults and children undergoing the Helicobacter pylori breath test. AB - The long-term biokinetics and dosimetry of carbon-14 were studied in nine adults and eight children undergoing carbon-14 urea breath test for Helicobacter pylori (HP) infection. The elimination of 14C via exhaled air and urine was measured with the liquid scintillation counting technique and with accelerator mass spectrometry. After the subjects had been given 110 kBq 14C-urea (children: 55 kBq) orally, samples of exhaled air were taken up to 180 days after administration and samples of urine were collected up to 40 days. Sixteen of the subjects were found to be HP-negative. In these subjects a total of 91.1%+/-3.9% (mean of adults and children +/- standard error of the mean) of the administered 14C activity was recovered. The majority of the administered activity, 88.3%+/ 6.2% in adults and 87.7%+/-5.0% in children, was excreted via the urine within 72 h after administration. A smaller fraction was exhaled. In adults 4.6%+/-0.6% of the activity was exhaled within 20 days and in children 2.6%+/-0.3%. Uncertainties in the biokinetic results are mainly due to assumptions concerning endogenous CO2 production and urinary excretion rate and are estimated to be less than 30%. The absorbed dose to various organs and the effective dose were calculated using the ICRP model for urea and CO2. The urinary bladder received the highest absorbed dose: in adults, 0.15+/-0.01 mGy/MBq and in children of various ages (7-14 years), 0.14-0.36 mGy/MBq. The findings indicate that an investigation with 14C-urea gives an effective dose to adults of 2.1+/-0.1 microSv (for 110 kBq) and to children of 0.9-2.5 microSv (for 55 kBq). From a radiation protection point of view, there is thus no reason for restrictions on even repeated screening investigations with 14C-urea in whole families, including children. PMID- 10369943 TI - Automatic segmentation of dynamic neuroreceptor single-photon emission tomography images using fuzzy clustering. AB - The segmentation of medical images is one of the most important steps in the analysis and quantification of imaging data. However, partial volume artefacts make accurate tissue boundary definition difficult, particularly for images with lower resolution commonly used in nuclear medicine. In single-photon emission tomography (SPET) neuroreceptor studies, areas of specific binding are usually delineated by manually drawing regions of interest (ROIs), a time-consuming and subjective process. This paper applies the technique of fuzzy c-means clustering (FCM) to automatically segment dynamic neuroreceptor SPET images. Fuzzy clustering was tested using a realistic, computer-generated, dynamic SPET phantom derived from segmenting an MR image of an anthropomorphic brain phantom. Also, the utility of applying FCM to real clinical data was assessed by comparison against conventional ROI analysis of iodine-123 iodobenzamide (IBZM) binding to dopamine D2/D3 receptors in the brains of humans. In addition, a further test of the methodology was assessed by applying FCM segmentation to [123I]IDAM images (5 iodo-2-[[2-2-[(dimethylamino)methyl]phenyl]thio] benzyl alcohol) of serotonin transporters in non-human primates. In the simulated dynamic SPET phantom, over a wide range of counts and ratios of specific binding to background, FCM correlated very strongly with the true counts (correlation coefficient r2>0.99, P<0.0001). Similarly, FCM gave segmentation of the [123I]IBZM data comparable with manual ROI analysis, with the binding ratios derived from both methods significantly correlated (r2=0.83, P<0.0001). Fuzzy clustering is a powerful tool for the automatic, unsupervised segmentation of dynamic neuroreceptor SPET images. Where other automated techniques fail completely, and manual ROI definition would be highly subjective, FCM is capable of segmenting noisy images in a robust and repeatable manner. PMID- 10369944 TI - Attenuation correction in whole-body FDG oncological studies: the role of statistical reconstruction. AB - Whole-body fluorine-18 fluoro-2-d-deoxyglucose positron emission tomography (FDG PET) is widely used in clinical centres for diagnosis, staging and therapy monitoring in oncology. Images are usually not corrected for attenuation since filtered backprojection (FBP) reconstruction methods require a 10 to 15-min transmission scan per bed position on most current PET devices equipped with germanium-68 rod transmission sources. Such an acquisition protocol would increase the total scanning time beyond acceptable limits. The aim of this work is to validate the use of iterative reconstruction methods, on both transmission and emission scans, in order to obtain a fully corrected whole-body study within a reasonable scanning time of 60 min. Five minute emission and 3-min transmission scans are acquired at each of the seven bed positions. The transmission data are reconstructed with OSEM (ordered subsets expectation maximization) and the last iteration is reprojected to obtain consistent attenuation correction factors (ACFs). The emission image is then also reconstructed with OSEM, using the emission scan corrected for normalization, scatter and decay together with the set of consistent ACFs as inputs. The total processing time is about 35 min, which is acceptable in a clinical environment. The image quality, readability and accuracy of uptake quantification were assessed in 38 patients scanned for various malignancies. The sensitivity for tumour detection was the same for the non-attenuation-corrected (NAC-FBP) and the attenuation-corrected (AC-OSEM) images. The AC-OSEM images were less noisy and easier to interpret. The interobserver reproducibility was significantly increased when compared with non corrected images (96.1% vs 81.1%, P<0.01). Standardized uptake values (SUVs) measured on images reconstructed with OSEM (AC-OSEM) and filtered backprojection (AC-FBP) were similar in all body regions except in the pelvic area, where SUVs were higher on AC-FBP images (mean increase 7.74%, P<0. 01). Our results show that, when statistical reconstruction is applied to both transmission and emission data, high quality quantitative whole-body images are obtained within a reasonable scanning (60 min) and processing time, making it applicable in clinical practice. PMID- 10369945 TI - Fluorodeoxyglucose positron emission tomography of soft tissue tumours: is a non invasive determination of biological activity possible? AB - Since musculoskeletal tumours comprise a large heterogeneous group of entities with different biological behaviour, clinical diagnosis of such lesions can be very difficult. The aim of this prospective study was to assess the usefulness of 2-[F-18]-fluoro-2-deoxy-d-glucose (FDG) positron emission tomography (PET) in the non-invasive evaluation of soft tissue tumours. One hundred and two patients with suspected soft tissue neoplasms were investigated by FDG-PET. The uptake of FDG was evaluated semiquantitatively by determining the tumour to background ratio (TBR). All patients underwent biopsy, resulting in the histological detection of 39 high-grade sarcomas, 16 intermediate-grade sarcomas, 11 low-grade sarcomas, 25 benign tumours, 10 tumour-like lesions such as spontaneous myositis ossificans (n = 6) and one non-Hodgkin lymphoma. All lesions except for two lipomas disclosed an increased FDG uptake. Sarcomas showed significantly higher TBR values than latent or active benign lesions (P<0.001) and aggressive benign lesions (P<0.05). Using a TBR cut-off level of 3.0 for malignancy, sensitivity of FDG-PET was 97.0%, specificity 65.7% and accuracy 86. 3%. From our data there are three main conclusions: (1) Except for patients with pseudotumoral myositis ossificans, lesions with a TBR >3 were sarcomas (91.7%) or aggressive benign tumours (8.3%). (2) Tumours with a TBR <1.5 were latent or active benign lesions, exclusively. (3) The group with intermediate TBR values (<3 and >1. 5) comprised primarily latent or active benign lesions, but also four aggressive benign tumours and two low-grade sarcomas. Our data suggest that FDG-PET represents a useful tool for the evaluation of the biological activity of soft tissue neoplasms. PMID- 10369946 TI - Clinical validation of the avidin/indium-111 biotin approach for imaging infection/inflammation in orthopaedic patients. AB - We report here the results of a validation study of the avidin/indium-111 biotin approach in patients with skeletal lesions. This study involved 54 patients with orthopaedic conditions: 20 patients with intermediate suspected osteomyelitis of the trunk, 19 patients with infection/inflammation of prosthetic joint replacements, and 15 patients with suspected osteomyelitis of appendicular bones. Avidin (3 mg) was injected as an i.v. bolus, followed 4 h later by 111In-biotin; imaging was acquired 30 min and 16-18 h after administration of 111In-biotin. Technetium-99m hexamethylpropylene amine oxime (99mTc-HMPAO)-labelled leucocyte scintigraphy was performed in 39/54 patients. The overall sensitivity of the avidin/111In-biotin scan was 97.7% (versus 88.9% for 99mTc-HMPAO leucocyte scintigraphy). While the diagnostic performance of avidin/111In-biotin scintigraphy was similar to that of 99mTc-HMPAO leucocyte scintigraphy in patients with prosthetic joint replacements or osteomyelitis of appendicular bones, the avidin/111In-biotin approach clearly performed better than 99mTc-HMPAO leucocyte scintigraphy in patients with suspected osteomyelitis of the trunk (100% sensitivity, specificity and accuracy versus 50% sensitivity, 100% specificity and 66.7% accuracy for 99mTc-HMPAO-leucocyte scintigraphy). These results demonstrate the feasibility of the avidin/111In-biotin approach for imaging sites of infection/inflammation in the clinical setting. Although no systematic advantages of avidin/111In-biotin scintigraphy were found versus 99mTc HMPAO leucocyte scintigraphy, the newer scintigraphic method is more practicable and involves lower biological risk for the operators. PMID- 10369947 TI - Allogenic bone graft viability after hip revision arthroplasty assessed by dynamic [18F]fluoride ion positron emission tomography. AB - The biological fate of allogenic bone grafts in the acetabular cavity and their metabolic activity after acetabular augmentation is uncertain but is most important for the stability of hip implants after hip revision arthroplasty. The aim of this study was to quantify regional bone metabolism after hip replacement operations. Dynamic [18F]fluoride ion positron emission tomography (PET) was used to investigate the metabolic activity of acetabular allogenic bone grafts and genuine bone, either 3-6 weeks (short-term group, n = 9) or 5 months to 9 years (long-term group, n = 10) after hip revision arthroplasty. Applying a three compartment model, the fluoride influx constant was calculated from individually fitted rate constants (Knlf) and by Patlak graphical analysis (Kpat). The results were compared with genuine cancellous and cortical acetabular bone of contralateral hips without surgical trauma (n = 7). In genuine cortical bone, Knlf was significantly increased in short- (+140.9%) and long-term (+100.0%) groups compared with contralateral hips. Allogenic bone grafts were characterised by a significantly increased Knlf in the short-term group (+190.9%) compared with contralateral hips, but decreased almost to the baseline levels of contralateral hips (+45.5%) in the long-term. Values of Knlf cor-related with the rate constant K1 in genuine (r = 0.89, P<0.001) and allogenic bone regions (r = 0.79, P<0.001), indicating a coupling between bone blood flow and bone metabolism in genuine bone as well as allogenic bone grafts. Kpat values were highly correlated with Knlf measurements in all regions. In conclusion, [18F]fluoride ion PET revealed the presence of an increased host bone formation in allogenic bone grafts early after hip revision arthroplasty. In contrast to genuine cortical bone, allogenic bone graft metabolism decreased over time, possibly due to a reduced ability to respond to the same extent as genuine bone to elevated metabolic demands after surgery. PMID- 10369948 TI - Serum bone alkaline phosphatase levels enhance the clinical utility of prostate specific antigen in the staging of newly diagnosed prostate cancer patients. AB - The aim of this study was to analyse the clinical utility of serum bone alkaline phosphatase (BAP) in addition to prostate-specific antigen (PSA) in the staging of newly diagnosed untreated prostate cancer patients. A prospective study was conducted, analysing serum BAP and PSA concentrations in 295 consecutive newly diagnosed untreated prostate cancer patients (T1-4, N0-1, M0-1b), 93 of whom had bone metastases on bone scan. The relationship of each marker with extent of bone disease, the influence of several clinical variables on both serum marker levels, the efficiency in predicting bone metastasis through receiver operating characteristic curves and, finally, the clinical utility in avoiding unnecessary bone scans were determined. Significant differences were found in the serum levels of both BAP and PSA between patients with and patients without bone metastases. Multiple regression analysis showed the extent of bone disease to be the only variable that influenced both serum levels. However, while serum BAP levels showed a statistical relationship with extent of bone disease, serum PSA levels did not. The best prediction of bone scan findings was obtained with the combination of both markers using a cut-off of 20 ng/ml, with positive and negative predictive values of 46.5% and 100%, respectively. This greater efficiency could permit 32.2% of initial bone scans to be avoided. False-positive and false-negative rates of BAP were 7.5% and 14%, respectively. This study suggests that serum BAP levels could play a complementary role in the diagnosis of bone metastasis in prostate cancer patients. This marker could provide useful clinical information on the degree of skeletal metastasis and constitute an easy way of enhancing the clinical utility of PSA. The addition of this marker to PSA in the initial evaluation could permit staging bone scan to be avoided at a PSA range of 10-20 ng/ml, with significant implications for cost saving. PMID- 10369949 TI - Reverse redistribution on thallium-201 single-photon emission tomography after primary angioplasty: a one-year follow-up study. AB - The prognostic significance of reverse redistribution (RR) on thallium-201 single photon emission tomography (SPET) images after acute myocardial infarction (AMI) has not been studied in detail. Moreover, RR data in patients treated with primary angioplasty are lacking. Fifty consecutive patients (including 40 men with a mean age of 54+/-11 years) with a first AMI were treated with primary angioplasty and followed up for 13+/-5 months for the following end-points: death, reinfarction and recurrent angina requiring revascularisation. Admission and peak creatine kinase myocardial enzyme (CKMB) and ejection fraction (EF) at discharge were studied as markers of myocardial damage. Thallium-201 stress redistribution SPET studies at 1 month were analysed using a 13-segment, 4-point scoring system. Segments showing a worsening of perfusion by at least 1 point on redistribution studies were defined as showing RR. RR was present in 13 (26%) patients (group 1) and absent in 37 (74%) (group 2). Both groups were comparable for age, sex, peak CKMB release, EF and Q-wave myocardial infarctions. TIMI flow 3 was obtained in 92% in group 1 and 95% in group 2 (P = 0.95). On admission, CKMB was significantly lower in group 1 (18+/-14 vs 44+/-41 U/l, P = 0.03). Also, segments showing reversible perfusion were significantly more frequent in group 2 (1/169 vs 57/481, P = 0. 01). During follow-up, no death occurred and the combined documented endpoint of reinfarction and recurrent angina requiring angioplasty or coronary artery bypass grafting was significantly more frequently reached in group 2 (0/13 vs 10/37, P = 0.046). In conclusion, RR is common (26%) after primary angioplasty for a first AMI and is associated with lesser myocardial damage on admission. Patients with RR rarely have reversible segments on 201Tl SPET and tend to have a favourable outcome after 1 year of follow-up. PMID- 10369950 TI - Thallium-201 right lung/heart ratio during exercise in patients with coronary artery disease: relation to thallium-201 myocardial single-photon emission tomography, rest and exercise left ventricular function and coronary angiography. AB - The aim of this study was to correlate lung thallium-201 uptake on exercise with 201Tl single-photon emission tomography (SPET) myocardial perfusion imaging, rest and exercise equilibrium radionuclide angiographic and coronary angiographic findings in patients with coronary artery disease (CAD) using a simple, reproducible lung/heart (L/H) ratio that would be easy to use in clinical practice. L/H ratio was defined on the anterior planar image obtained during exercise 201Tl SPET acquisition as the mean counts per pixel in an entire right lung field region of interest divided by the mean counts per pixel in the hottest myocardial wall region of interest. We studied 103 patients. Fifty-nine patients (group I) with <5% likelihood of CAD were used as a reference group. In 44 CAD patients (group II), L/H ratio was compared with 201Tl SPET, radionuclide angiographic and coronary angiographic variables. The group I L/H ratio of 0.35+/ 0.05 (mean +/-1 SD) was significantly lower (P<0.001) than the group II L/H ratio of 0. 45+/-0.10. An L/H ratio >0.45 (mean + 2 SD in group I) was considered abnormal. In group II, L/H ratio showed a significant correlation with stress and rest 201Tl perfusion defect size (r = 0. 39 and r = 0.42, P<0.01, respectively), but not with extent of ischaemic myocardium. The mean L/H ratio was 0.41+/-0.10 in patients with one-vessel disease (n = 15), 0.46+/-0.08 in those with two vessel disease (n = 17) and 0.47+/-0.12 in those with three-vessel disease (n = 12), but no significant difference was found between the three subgroups. L/H ratio showed a significant inverse relation with rest and exercise left ventricular ejection fraction (r = -0.37 and r = -0.50, P<0.05 and P<0.001, respectively). Using stepwise multiple regression analysis, exercise left ventricular ejection fraction and previous history of hypertension were the sole two variables independently predictive of the L/H ratio. In conclusion, although lung thallium uptake is usually found to correlate with extent and severity of CAD, increased L/H ratio should primarily be considered as a marker of exercise induced left ventricular systolic and perhaps diastolic dysfunction, probably independent of the underlying cardiac disease. PMID- 10369951 TI - Potential of iodine-123 metaiodobenzylguanidine single-photon emission tomography to detect abnormal functional status of the pulmonary neuroadrenergic system in irradiated lung. AB - The potential of iodine-123 metaiodobenzylguanidine (MIBG) to detect functional abnormalities of the pulmonary neuroadrenergic system (PNS) in irradiated lung areas (ILAS) was preliminarily explored using single-photon emission tomography (SPET). The subjects included five healthy subjects and a total of 31 patients with peripheral-type lung cancer treated by radiation; 15 patients (group A) had received a dose of less than 36 Gy (mean +/- SD: 28.2 +/-6.2 Gy), and 16 patients (group B) had received a higher dose (mean +/- SD: 51.2 +/- 3.5 Gy) at the time of examination. MIBG SPET scans acquired 15 min and 3 h after injection were used to measure the MIBG uptake ratio (count ratio of the ILA to the contralateral non ILA) and the clearance rate [percentage of (early counts - delayed counts)/early counts] from the ILAs without noticeable abnormal opacities on chest computed tomography scan. Lung perfusion changes were also assessed by technetium-99m macroaggregated albumin SPET. By contrast to the homogeneous MIBG uptake in the lungs of the healthy subjects, MIBG uptake was folcally decreased in correspondence with the ILAs in all patients, including 11 patients (73.3%) of group A with relatively preserved lung perfusion. The reduction MIBG uptake was significant (P<0.0001), and the MIBG clearance rate from the ILAs was also significantly faster than the clearance rates from the normal lungs and contralateral non-ILAs (both P<0.01). Group B patients showed significantly lower MIBG uptake and faster clearance from the ILAs than group A patients (P<0. 001 and P<0.05, respectively), although there was no significant difference in the clearance from the non-ILAs. Overall, MIBG uptake/clearance from the ILAs correlated significantly with the radiation dose in the 31 patients (r = -0.656; P<0.0001 and r = 0. 387; P<0.05, respectively). Perfusion changes were inversely correlated with the clearance from the ILAs (r = -0.432, P<0.05), but did not correlate with MIBG uptake. These preliminary results suggest that MIBG may have the potential to be a marker of abnormal functional status of the PNS produced by irradiation and may facilitate investigation of irradiation lung injury independently of morphological or lung perfusion changes. PMID- 10369952 TI - Influence of high-energy photons on the spectrum of iodine-123 with low- and medium-energy collimators: consequences for imaging with 123I-labelled compounds in clinical practice. AB - Using iodine-123 labelled radiotracers, the presence of 2.5% high-energy photons causes image deterioration due to increased scatter. To investigate the influence of these photons on image quality, we measured the spectrum of 123I with a medium energy (ME), a low-energy all-purpose (LEAP) and a low-energy high-resolution (LEHR) collimator. Even in air, using low-energy collimators a high baseline activity was observed over the total energy detection range of the gamma camera. The 159-keV photopeak to scatter activity ratio fell from 5.9 for ME to 3.6 and 2.9 for LE collimators. Acquisition of images with LEHR collimators with energy windows set at 159 keV and 500 keV demonstrated that the 159-keV LEHR image is a combination of the ME image of the object and of the LEHR 500-keV image. Because of their important septal penetration and greater geometric detection efficiency compared with the 159-keV photons of 123I, the contribution of high-energy photons is dependent on the source-detector distance. For a small source placed in air, the scatter to photopeak activities varied from 17.4% at 80 cm to 37.8% at 5 cm distance from an LEHR collimator. Considering only the scatter problem, ME collimators are the best choice for 123I studies. When using LE collimators for high-resolution tomography with 123I-labelled compounds, scatter contribution from high-energy photons has to be corrected for quantitative analysis or when dual-isotope studies are performed, whether or not these studies are acquired simultaneously. PMID- 10369953 TI - Radioiodine uptake in inactive pulmonary tuberculosis. AB - Radioiodine may accumulate at sites of inflammation or infection. We have seen such accumulation in six thyroid cancer patients with a history of previously treated pulmonary tuberculosis. We also review the causes of false-positive radioiodine uptake in lung infection/inflammation. Eight foci of radioiodine uptake were seen on six iodine-123 diagnostic scans. In three foci, the uptake was focal and indistinguishable from thyroid cancer pulmonary metastases from thyroid cancer. In the remaining foci, the uptake appeared nonsegmental, linear or lobar, suggesting a false-positive finding. The uptake was unchanged, variable in appearance or non-persistent on follow-up scans and less extensive than the fibrocystic changes seen on chest radiographs. In the two patients studied, thyroid hormone level did not affect the radioiodine lung uptake and there was congruent gallium-67 uptake. None of the patients had any evidence of thyroid cancer recurrence or of reactivation of tuberculosis and only two patients had chronic intermittent chest symptoms. Severe bronchiectasis, active tuberculosis, acute bronchitis, respiratory bronchiolitis, rheumatoid arthritis-associated lung disease and fungal infection such as Allescheria boydii and aspergillosis can lead to different patterns of radioiodine chest uptake mimicking pulmonary metastases. Pulmonary scarring secondary to tuberculosis may predispose to localized radioiodine accumulation even in the absence of clinically evident active infection. False-positive radioiodine uptake due to pulmonary infection/inflammation should be considered in thyroid cancer patients prior to the diagnosis of pulmonary metastases. PMID- 10369954 TI - Economic evaluation studies in nuclear medicine: the need for standardization. AB - The guidelines for publishing economic evaluations require a statement of the economic importance of the analysis and the viewpoint from which it has been carried out, as well as specification of at least two alternative programmes or interventions, the form of economic evaluation, the outcome measure, the method of costing, the time horizon and adjustment for timing of costs and benefits (e.g. by a discount factor), and the allowance for uncertainties (e.g. by implementation of a sensitivity analysis). The decision analysis can be based on clinical trial data, on retrospective or administrative databases, or on modelling. The choice of outcome measures is the key issue in an economic evaluation. In cost-effectiveness analysis, benefits are usually measured in natural units. This is the form of economic evaluation most frequently used in nuclear medicine. Endpoints of effectiveness applied in studies in this field have been procedures avoided, procedures initiated, cardiac events, survival probability, morbidity, quality of life and protracted or failed surgical procedures. In other instances, surrogate endpoints have been used such as metastases detected, staging, viability or tumour response. This, however, limits comparability of cost-effectiveness considerably, as proof of a change in the health outcome cannot be obtained. Measures of utility such as QALYs (quality adjusted life years) have so far only been applied for decision tree analysis. Useful examples of economic evaluation studies in nuclear medicine are presented here for fluorodeoxyglucose positron emission tomography (FDG-PET) in the preoperative staging of non-small cell lung cancer, for FDG-PET in differentiating indeterminate solitary pulmonary nodules, for somatostatin receptor scintigraphy in detecting metastases of carcinoid tumours, for routine preoperative scintigraphy with sestamibi in patients with parathyroid adenoma, for periodic measurement of thyroid-stimulating hormone in detecting mild thyroid failure, for diagnostic algorithms including a lung scan in patients with suspected pulmonary embolism, for myocardial perfusion imaging as an incremental prognostic factor in patients with coronary artery disease, and for the use of radioiodine as first-line therapy of Graves' hyperthyroidism and of toxic nodular goitres. Further evaluations of effectiveness or utility should be carried out within a multidisciplinary framework to ensure that nuclear medical procedures are included in the general management guidelines. PMID- 10369955 TI - Leu343Phe substitution in the Malx3 protein of Saccharomyces cerevisiae increases the constitutivity and glucose insensitivity of MAL gene expression. AB - To utilise maltose as a carbon source Saccharomyces cerevisiae needs one or more functional MAL loci that contain the MALx1 gene encoding maltose permease, MALx2 encoding maltase, and MALx3 encoding a transcriptional activator. Maltose causes a rapid MALx3-dependent induction of MAL gene transcription, and glucose represses this activation via Mig1p. A MALx3 gene conveying high MAL gene expression in the absence of maltose in a malx3 laboratory mutant strain has been isolated from baker's yeast. The construction of hybrid genes between the isolated gene and a highly regulated MALx3 gene showed that constitutivity was the result of multiple amino-acid alterations throughout the structural gene. The combined effect of these amino-acid alterations was shown to be stronger than the sum of their individual effects on constitutivity. Analysis in glucose-repressed conditions confirmed that increased MALx3 transcript levels increased the glucose insensitivity of MAL gene expression but did not affect constitutivity. Analysis of four mutations between aa 343 and 375, lying within a proposed negative regulatory domain, showed that the single mutation of Leu343Phe increased the glucose insensitivity of MAL gene expression by 30-fold. These results demonstrate that not only Mig1p modulation of MALx3 expression, but also the MALx3 protein structure, is involved in the glucose-insensitive expression of the MAL genes. PMID- 10369956 TI - Strand interruptions confer strand preference during intracellular correction of a plasmid-borne mismatch in Saccharomyces cerevisiae. AB - Site-directed mutagenesis was used to construct yeast centromere plasmids in which a strand nick or gap could be placed 5' or 3', on either strand, to a reporter gene (SUP4-o) carrying defined base mismatches. The plasmids were then transformed into yeast cells and the direction and efficiency of mismatch repair were assayed by scoring colouring of the transformant colonies. Strands that were nicked were consistently corrected more often than intact strands, but the effect was very small. However, placement of a small gap at the same positions as the nicks resulted in a marked increase in selection for the gapped strand and an enhanced efficiency of mismatch repair. Both the preference for the gapped strand and correction of the mismatch were offset by deletion of the mismatch repair gene PMS1. Together, the results suggest that strand interruptions can direct intracellular mismatch correction of plasmid-borne base mispairs in yeast. PMID- 10369957 TI - DNA-binding factors assemble in a sequence-specific manner on the maize mitochondrial atpA promoter. AB - The maize mitochondrial atpA promoter has been well-characterized using in vitro transcription. The functional elements of this promoter comprise a central domain extending from -7 to +5 relative to the transcription start site, and an upstream domain of 1-3 bp that is purine-rich and centered around positions -11 to -12. As a first step in characterizing the transcriptional machinery, exonuclease-III mapping (toeprinting) was used to map the borders of DNA-protein interactions using either a 107-bp wild-type template or transcriptionally-inactive templates containing linker-scanning mutations. These experiments revealed that, with a wild-type promoter, protein factors occupy as much as 36 bp, from positions -20 to +16 relative to the transcription initiation site. Protein-binding patterns were altered when the linker-scanning mutants were used, suggesting that either the number or conformation of DNA-binding proteins could account for their inability to promote transcription initiation. PMID- 10369958 TI - ATP synthase 5' untranslated regions are specifically bound by chloroplast polypeptides. AB - Expression of the large ATP synthase gene cluster in spinach (Spinacia oleracea) chloroplasts is regulated at the post-transcriptional level. RNA stability and the translational efficiency of some chloroplast transcripts have been shown to be regulated through RNA-protein interactions in the 5' untranslated region (5' UTR). In this report we show that spinach chloroplast extracts contain polypeptides that specifically interact with the 5' UTRs of three of the four genes in the large ATP synthase gene cluster. A subset of binding polypeptides may be gene-specific, although at least one appears to be a more general chloroplast RNA-binding protein. We hypothesize that these RNA-protein interactions may affect the expression of this gene cluster from two perspectives. The first would be at a gene-specific level, which could serve to control the stoichiometry of ATP synthase subunits. The second would be a more global effect, which may adjust the abundance of the entire ATP synthase complex in response to environmental or developmental cues. PMID- 10369959 TI - Pleiotropic effects of a nuclear restorer-of-fertility locus on mitochondrial transcripts in male-fertile and S male-sterile maize. AB - Cytoplasmic male sterility (CMS) is encoded by the plant mitochondrial genome and can be reversed by nuclear restorer-of-fertility(Rf) alleles. In the CMS-S system of maize, reproductive failure and fertility restoration are gametophytic, occurring during the starch-filling stages of pollen development. Transcripts of the CMS-S-associated mitochondrial open reading frames (orf355 and orf77) are present from the early stages of microspore development through the aborted pollen stage. To investigate the molecular basis of fertility restoration, we compared mitochondrial-transcript accumulation in aborting CMS-S pollen and in CMS-S pollen restored to fertility by the Rf3 nuclear allele. In the presence of the Rf3 allele, novel, shorter transcripts of the orf355-orf77, cob and atp6 mitochondrial genes were created, and the relative abundance of larger transcripts was decreased for each of these loci. The altered transcript patterns cosegregated with male fertility conditioned by the Rf3 allele. The novel cob and atp6 transcripts were also observed in leaf-tissues of both normal and S cytoplasm plants carrying the Rf3 allele. These observations support the hypothesis that the Rf3 allele encodes, or regulates, a modifier of mitochondrial transcript (Mmt) activity that affects both CMS and essential mitochondrial gene transcripts. PMID- 10369960 TI - Comparison of ribosomal DNA ITS regions among geographic isolates of Cenococcum geophilum. AB - Cenococcum geophilum is an ecologically important mycorrhizal fungus with a global distribution and a wide host range. It has been difficult to study since it forms only sterile mycelia and, occasionally, sclerotial bodies. Because of its lack of morphological variability, its taxonomy and phylogenetic origins have until recently remained unclear. To better understand the genetic variation and environmental adaptability of C. geophilum, a molecular phylogeny was constructed based on the nuclear ribosomal DNA internal transcribed spacers (ITS1 and ITS2) of 69 isolates from various hosts and habitats. The results suggest DNA sequence conservation in the ITS regions. Considering its broad geographic and host range, this ITS conservation was unexpected. Our data imply that the ITS2 region is under evolutionary pressure to maintain the RNA secondary structure (similar to the pressure on the CgSSU introns) involved in the post-transcriptional processing of rRNA. Also, C. geophilum has very short ITS regions, thus limiting the number of mutable sites. This limited ITS variability suggests a recent radiation of C. geophilum, having been geographically distributed by a variety of efficient processes. C. geophilum appears to be a single taxonomic entity, possibly a single species. Therefore, it is an extremely adaptable, as well as ecologically valuable, taxon. PMID- 10369961 TI - An exceptional group-I intron-like insertion in the SSU rDNA of lichen mycobionts. AB - An exceptional group-I intron-like insertion at position 940 of the nuclear small subunit rDNA is found in lichen mycobionts of the families Parmeliaceae and Lecanoraceae. This shared insertion site is exceptional as it follows a G. Although several features of the self-splicing Tetrahymena intron are missing, the conserved structure of the presumed core region indicates that the new intron like insertion, which is missing in mature transcripts, is not part of a silenced ribosomal repeat. It is unlikely that the new insertion is horizontally transferred from the adjacent position 943. A phylogenetic analysis indicates congruence with lichen phylogeny and suggests that the insertion has been vertically inherited. PMID- 10369962 TI - Trans-splicing and cis-splicing in the colourless Euglenoid, Entosiphon sulcatum. AB - The colourless Euglenoid Entosiphon sulcatum is thought to have diverged before the symbiotic event which gave rise to the photosynthetic Euglenoid species as Euglena gracilis. We have isolated genes encoding spliced leader-sequence RNA (SL RNA) and we show that pre-mRNAs are matured via a trans-splicing reaction in E. sulcatum, as in the case of E. gracilis. The 2.5-kb repeated DNA fragment which encodes the SL-RNA gene also encodes a 5S rRNA gene as well as the genes for the small nuclear (sn) RNAs U1, U2 and U5. The presence of snRNA U1 indicates that the classical cis-splicing mechanism also exists in E. sulcatum. In addition, we show that the E. sulcatum beta-tubulin gene has the intron borders GU-AG, typical of spliceosome-matured introns which have not yet been found in E. gracilis. The probable origins of trans- and cis-mechanisms in Euglenoids are discussed. PMID- 10369963 TI - Identification and characterization of brt1, a gene down-regulated during B regulated development in Schizophyllum commune. AB - To identify genes regulated by the B mating-type genes a differential RNA display was established for the homobasidiomycete Schizophyllum commune and a gene, brt1, was identified. The mRNA concerned is highly abundant in monokaryotic mycelia and is down-regulated in matings in which either the B genes alone or else both the A and B genes are different between the mates. This places the gene brt1 under the control of the pheromone response system of S. commune encoded by the B mating type loci. Sequence analysis revealed similarity to a novel protein family, two members of which have been shown to inhibit translational re-initiation. PMID- 10369964 TI - PaGrg1, a glucose-repressible gene of Podospora anserina that is differentially expressed during lifespan. AB - A loss-of-function mutation in Grisea, a gene of Podospora anserina that was previously shown to code for a copper-modulated transcription factor, leads to a significant increase in lifespan. In an attempt to identify and to isolate potential target genes controlled by GRISEA, a RT-differential-display screen was performed. This approach resulted in the identification of a gene, PaGrg1, that is differentially expressed in the wild-type and in the long-lived grisea mutant. In the mutant, transcript levels of PaGrg1 were found to be much lower than in the wild-type even if copper was added to the growth medium in amounts that revert the phenotype of this copper-uptake mutant to wild-type characteristics. PaGrg1 is a discontinuous gene with a single intron and encodes a protein of 71 amino acids sharing a high degree of sequence identity (65%) with the developmentally regulated, catabolite-repressed grg-1 gene of Neurospora crassa. Transcription of PaGrg1 increases upon carbon starvation indicating that PaGRG1 represents a putative stress protein. Transcript levels of PaGrg1 were found to increase during aging in both the wild-type strain and the long-lived mutant. However, even in the senescent stage of grisea, they are much lower than in juvenile cultures of the wild-type strain. The data suggest that threshold transcript levels of PaGrg1, and/ or additional unidentified genes which are controlled by GRISEA and which are subject to catabolite repression, are significantly involved in lifespan control. This conclusion is supported by the finding that, in contrast to the wild-type, the lifespan of grisea does not increase when cultures are grown on non-repressible carbon sources. PMID- 10369966 TI - The growth industry of the nineties. AB - This article critically reviews the current state of knowledge relating to outcome following decompressive surgery for lumbar spinal stenosis. Non-biased reviews of outcome are rare, follow-up times are not uniform and generally short, surgical techniques are not standardized. Rationale regarding the efficacy of concomitant spinal fusion is unconvincing. A procedure that is becoming ever more common in the neurosurgical armamentarium appears to be poorly described with respect to outcomes. PMID- 10369965 TI - Aa-Pri2, a single-copy gene from Agrocybe aegerita, specifically expressed during fruiting initiation, encodes a hydrophobin with a leucine-zipper domain. AB - The Aa-Pri2 gene, specifically expressed during basidiocarp differentiation of the mushroom Agrocybe aegerita, was cloned. Sequence analysis showed a 525-nt ORF interrupted by three introns (51, 48, 54 bp). One TATA box (-94), two CAAT boxes (-382, -398) and one CT-rich motif (-149) were present. The transcription start point was located by primer extension at position -60. The hydrophobic Aa-PRI2 protein (123 aa) possessed a putative signal peptide (aa1-aa18), eight cysteine residues characteristic of the hydrophobin proteins, and one putative leucine zipper domain (aa50-aa71). Hybridization of the genomic DNA of several edible mushrooms with the Aa-Pri2 cDNA, showed a 2.7-kb DNA fragment highly similar to the Aa-Pri2 gene in the species Agrocybe chaxingu; sequences showing slight hybridization signals were detected in Pleurotus eryngii, Lentinula edodes and Agaricus bisporus. PMID- 10369967 TI - Do cerebral arteriovenous malformations recur after angiographically confirmed total extirpation? AB - Cerebral arteriovenous malformations (AVMs) are thought to result from a failure of embryogenesis in the otherwise normal differentiation of primordial vascular channels into mature arteries, capillaries, and veins. Although these are essentially congenital vascular malformations, marked enlargement and/or recurrence of cerebral AVMs has been reported in the recent literature. Using MEDLINE (1966- 1998), we searched the recurrence of cerebral AVMs and analyzed all reported recurrent cases after total surgical extirpation and negative postoperative angiogram, and discussed the proposed mechanisms of the recurrence of cerebral AVMs. A thorough literature survey disclosed only 12 documented recurrent cases (9 were documented in English and 3 in Japanese), which shows the rarity of the recurrence of cerebral AVMs, although the actual rate of recurrence is not known because of the lack of routine long-term follow-up. The location of recurrent cerebral AVMs was the cerebral hemisphere, and initial presentation was hemorrhage in all cases. Recurrence occurred in patients under 20 years of age in 9 of 11 cases, which implies the propensity of recurrence of cerebral AVMs in immature brain vasculature. There are no definite proven mechanisms to explain why congenital anomalies such as cerebral AVMs recur after total extirpation, but recently two plausible mechanisms have been proposed. One is angiogenesis disregulated by vas-cular endothelial growth factor (VEGF) and the other is a new anatomical entity, 'hidden compart-ments'. Although VEGF is one of the main angiogenetic factors and its important role in fetal brain and pathological neovascularization has been reported, the synthesis of VEGF might be insufficient to explain the recurrence of cerebral AVMs because VEGF-positive staining is also found in nonrecurrent patients. Hidden compartments are angiographically unfilled compartments, in spite of an adequate examination, which may be located within, contiguous with, or relatively far from the angiographically demonstrated AVM. Although it might explain unsolved clinical phenomena such as regrowth, recurrence, and per- or postoperative unanticipated bleeding and brain swelling, the existence of hidden compartments should be proved by high-resolution radiological examinations or during operations. The presence of recurrent cerebral AVMs after complete extirpation by modern microsurgical techniques indicates that cerebral angiography in the early postoperative stage, the golden standard to assess the disappearance of cerebral AVMs, is not sufficient to eliminate the risk of hemorrhage, and careful long-term follow-up studies should be planned. PMID- 10369968 TI - Subarachnoid haemorrhage of unknown aetiology: what next? AB - Subarachnoid haemorrhage (SAH) is often associated with negative cerebral angiography. Following the exclusion of other causes, a patient may be suspected of harbouring an occult intracranial aneurysm, with risk of recurrent bleeding and death. These patients are often identified on the basis of clinical presentation and computed tomography (CT) findings, and require expeditious further investigation if morbidity and mortality are to be minimized. Currently available options include repeated cerebral angiography, surgical exploration, and the newer technologies of computed tomography angiography (CTA) and magnetic resonance angiography (MRA). We review these options, based on current literature, with particular emphasis on the expanding roles of CTA and MRA. A multimodality management protocol is proposed, with decisions based on clinical urgency, patient progress and the natural history of aneurysmal SAH, particularly vasospasm and aneurysm thrombosis. PMID- 10369969 TI - Surgical treatment of dumbbell neurinomas of the cervical spine. AB - Cervical neurinomas ex-tending through the intervertebral foramen are uncommon. They raise difficult problems of surgical management. The few papers dealing with surgical technique for removal of these tumors are reviewed. The surgical approach has to be chosen from posterior, anterior, and anterolateral routes. Analysis is essentially devoted to the radicality of tumor resection, nerve root preservation, relation to the vertebral artery, and compromise of spinal stability. PMID- 10369970 TI - Proton magnetic resonance spectroscopy. AB - Proton magnetic resonance spectroscopy (MRS) permits in vivo determination of biochemical parameters within brain tissue, utilizing the same magnetic resonance (MR) scanner and head coil that are utilized for conventional MR imaging. This technology has been evolving and improving over the past decade, with most of the current published work based on the measurement of rather large single voxels, utilizing variable echo times, aimed at characterizing changes in brain tumors, demyelinating diseases, abscesses, and metabolic diseases. Future work will utilize multiple-voxel techniques so that volumes of tissue can be examined with smaller voxels in reasonable acquisition times, providing a greater understanding of the metabolite composition of the brain, especially as more and more metabolites are identified within the spectra. PMID- 10369971 TI - Radiosurgery for pituitary adenoma. AB - The role of irradiation in the management of pituitary adenomas is not well defined. Nevertheless, patients with residual or recurrent tumours have been treated with conventional external-beam radiotherapy and more recently with high precision stereotactic techniques of stereotactic radiosurgery (SRS) and stereotactic radiotherapy (SRT). We review some of the recently published articles on the efficacy and toxicity of SRS in the light of the current literature describing the results of conventional radiotherapy. While the general perception is that single-fraction SRS is more effective and less toxic than fractionated radiotherapy and in hormone-secreting tumours may produce a faster decline in elevated hormone levels, the available evidence suggests higher toxicity than seen with fractionated treatment without the reassurance of long term tumour control. There is also no convincing evidence for more rapid reduction of elevated hormones. For the treatment of larger non-spherical pituitary adenomas, it may be appropriate to explore a potentially safer high precision technique of fractionated conformal stereotactic radiotherapy (SCRT). In conclusion, there is currently little justification for the routine use of single-fraction SRS for the treatment of the majority of patients with benign pituitary adenomas. PMID- 10369972 TI - Medulloblastoma - late outcome. AB - Long-term survivors of medulloblastoma do occur, even in high-risk groups. Infants and toddlers surviving past the age of 21 years can expect significant intellectual impairment if their radiation therapy was given at approximately 2 years of age. If radiation therapy is delayed or avoided, most of those who survive long term are in a regular school classroom. All of the latter had complete tumor resection. Sixty-two percent of children who received conventional radiation therapy (mean 24 Gy) developed primary hypothyroidism. Tumor markers and improved understanding of tumor biology may lead to more effective surveillance monitoring. PMID- 10369973 TI - Meningiomas in childhood. AB - Meningiomas are rare intracranial neoplasms in the pediatric population. Most of the reports concerning these tumors have stressed some distinguishing features comparable with the adult counterpart. Besides the lower incidence, the nearly equal sex distribution, the relatively common infratentorial location and the frequent development within the ventricular system have been emphasized. A poor prognosis of pediatric meningiomas has also been claimed by authors who have suggested that this could be owing to a possibly more aggressive behavior, accounting for the huge size that these tumors have usually reached at diagnosis, the difficult surgical excision, and the higher recurrence rate. The analysis of recent series challenges the concept that pediatric meningiomas do actually bear a worse prognosis than similar tumors occurring in older patients. Improved surgical techniques have in fact resulted in higher percentages of complete tumor removal. Better histological delineation has allowed the identification of highly aggressive meningeal neoplasms, which in the past were not differentiated properly, thus contributing to the apparent higher incidence of recurrent tumors in the pediatric population. When the studies are limited to the "classical" form of meningioma, it becomes apparent that pediatric meningiomas do not behave more aggressively than meningiomas in adult patients. PMID- 10369974 TI - Latex allergy in spina bifida patients. AB - Latex allergy has become a major problem in children with spina bifida, who need to undergo many major aggressive diagnostic and therapeutic procedures. Latex allergy is increasing in medical and surgical practice. Although early reports of latex allergy date from 1927, only over the last decade there has been more attention paid to latex allergy. This is due to an increasing number of reported cases of mild to fatal adverse reactions to latex. Risk groups have been identified; among these are patients undergoing multiple surgeries such as those with spina bifida. In this critical review, we aim to emphasize some aspects of the current management of surgical patients with latex allergy. PMID- 10369976 TI - Papers reviewed in this issue. PMID- 10369977 TI - Publications scanned for pertinent articles. PMID- 10369978 TI - Tumor diagnosis in the adult liver transplant candidate. AB - Hepatic transplantation has emerged as a potentially curative treatment of certain malignant hepatic neoplasms such as hepatocellular carcinoma, bile duct carcinoma, fibrolamellar hepatocellular carcinoma, metastases from neuroendocrine tumors, and epithelioid hemangioendothelioma. In the early years of hepatic transplantation, there was great enthusiasm to cure patients with unresectable hepatobiliary malignancy. This early enthusiasm was tempered by the unfavorable outcome of transplantation in advanced cases of malignancy and the organ-donor shortage. Presently, patients have to be selected with predictable likelihood for long-term survival. Pre-transplantation imaging is indispensable for detection, characterization, staging, and surgical road-mapping before the procedure. The present article focuses on the role of imaging modalities in these different aspects of preoperative assessment. PMID- 10369979 TI - Screening applications for MRI in the detection of upper abdominal disease: comparative study of non-contrast-enhanced single-shot MRI and contrast-enhanced helical CT. AB - PURPOSE: To compare the value of 'push-button' single-shot non-contrast-enhanced MRI and contrast-enhanced helical CT for detection of upper abdominal disease. METHODS: In 120 patients, images obtained with non contrast-enhanced single-shot MRI (T2: double echo HASTE, and T1: turbo FLASH) and contrast-enhanced helical CT were compared. Lesions or abnormalities were divided in 8 anatomical categories (1: liver; 2: pancreatobiliary; 3: kidney/adrenal gland; 4: retroperitoneum; 5: vascular; 6: spleen; 7: gastrointestinal tract and peritoneum; 8: base of thorax) and classified as follows: 2: seen at MRI only; 1: better seen at MRI; 0: no difference; -1: better seen at CT; -2: seen at CT only. Also recorded were the 'door-to-door' examination times. RESULTS: Of a total of 629 abnormalities, 594 were detected at MRI (94 %) and 536 at CT (85 %). CT offered better results in two categories only: retroperitoneum (mean score: -0.68) and vascular (mean score -0.87). Mean examination times were 19 min for CT and 14.8 min for MRI. CONCLUSION: Unenhanced single-shot MRI is a valuable first step of a comprehensive upper abdominal MR exam and may even be the final step in many patients. PMID- 10369981 TI - Pathways of lymph node involvement in upper abdominal malignancies: evaluation with high-resolution CT. AB - The aim of this study was to enhance our understanding of the pathways of lymphatic spread of primary carcinomas in the upper abdomen by recognizing the development, configuration, and frequency of nodal enlargement in discrete anatomic regions. The study included 417 patients with histologically confirmed carcinomas (CC) of the stomach (n = 267), liver (n = 98), gallbladder (n = 25), and bile ducts (n = 27). All patients were studied by high-resolution CT and tumor extension to the lymph nodes of the subperitoneal space was clearly identified in 59 patients [33 with CC of the stomach, 8 with CC of the gallbladder, 3 with CC of the bile ducts, and 15 with hepatocellular carcinoma (HCC)]. In 47 of 59 patients this extension was confirmed by surgery or aspiration biopsy. Three discrete anatomic groups of lymph nodes were recognized producing a relatively distinct CT configuration when involved: (a) the hepatoduodenal seen in 49 patients; (b) the peripancreatic demonstrated in 33 patients; and (c) the aortocaval recognized in 16 patients. These groups of lymph nodes can be seen individually involved or in combination. Recognition of involvement of these nodes is important for correct diagnosis and staging of upper abdominal malignancies. The development of this involvement follows the natural flow of lymph via the lesser omentum to the retroperitoneal space. PMID- 10369980 TI - New sonographic imaging observations in focal pancreatitis. AB - The imaging findings that ultrasonographically differentiate focal acute pancreatitis (FAP) from a malignant lesion of the pancreas are described. Focal acute pancreatitis is ultrasonographically (US) characterized as a hypoechoic, homogeneous, localized, subsegmental, non-expansive and diffusely demarcated lesion located mostly in the head of the pancreas. It could not be visualized using CT. Endoscopic retrograde cholangiopancreatography (ERCP) performed in 13 of the 32 patients, showed chronic pancreatitis. Focal acute pancreatitis disappeared in 1-6 months at US follow-up. The clinical diagnoses were acute pancreatitis in 11 patients, chronic pancreatitis in 12 patients, biliary disease in 5 patients, hepatopathia in 1 patient while the diagnosis was unknown in 2 patients. No patient developed any pancreatic cancer during a median of 85 months of follow-up. In conclusion, the present data indicate that patients with FAP at US, without any focal lesion seen on either CT or ERCP, have a benign pancreatic lesion, which resolves in 1-6 months; thus, such patients probably do not need any further investigation or follow-up at all. PMID- 10369982 TI - Double-contrast magnetic resonance examination of ulcerative colitis. AB - The aim of our work was to propose a double-contrast magnetic resonance examination (DCMRE) in the follow-up of ulcerative colitis (UC), comparing this new technique with X-ray double-contrast barium enema (DCBE). After preparation with colon-cleansing regimen used for DCBE, six UC patients and six control subjects underwent a 1.5-T examination: supine position, coronal and axial fat spectral-saturation breath-hold gradient-echo T1-weighted sequences after intravenous hypotonization and 1500-2000 cc air insufflation. Without evacuating the primarily insufflated air, the same images were acquired after endorectal administration of negative superparamagnetic contrast agent (600 cc) and intravenous administration of positive paramagnetic contrast agent (0.2 mmol/kg). All patients had undergone DCBE in the four preceding weeks. We found significant increase in wall thickness of UC affected vs apparently unaffected segments (p = 0.0425) and vs CG (p = 0.0447), significant increase in enhancement percent of UC affected vs apparently unaffected segments (p = 0.0161) and vs CG (p = 0.0185), and no significant difference for enhancement percent of UC unaffected segments vs CG. DCMRE and DCBE localized the UC extension at the same sites in all patients. Double-contrast MR examination time was 20-30 min. This new method could be used in follow-up of UC patients. PMID- 10369983 TI - US-guided gallbladder aspiration: a new diagnostic method for biliary fascioliasis. AB - Fasciola hepatica is a trematode which is found worldwide. The diagnosis is usually delayed because the disease is relatively rare and the parasite or its eggs must be shown in bile samples for verification. We report three cases in which the diagnosis of fascioliasis was established by simple US-guided aspiration of the gallbladder. This new diagnostic method is less invasive, safe, and easy compared with the conventional endoscopic methods. PMID- 10369984 TI - Atypical MRI presentation of a small splenic hamartoma. AB - Hamartomas of the spleen usually appear isointense on T1-weighted MR images and hyperintense on T2-weighted images. We describe a histologically proven case which presented as a small (2.5 cm) focal mass isointense to splenic parenchyma on T1-weighted images and hypointense on both turbo-spin-echo T2 and short T1 inversion recovery images. Dynamic MRI revealed a delayed enhancement during the arterial phase becoming isointense and prolonged on subsequent images. This prolonged enhancement has previously been described as a characteristic pattern in these tumours. The lack of oedema and necrosis and the presence of fibrous tissue in the hamartoma at histopathology likely account for the low signal intensity on all sequences. PMID- 10369985 TI - Acute hepatobiliary tuberculosis: a report of two cases and a review of the literature. AB - Two cases of hepatobiliary tuberculosis are described. Case one, the macronodular type of hepatic tuberculosis, presented as pyrexia of unknown origin and was eventually diagnosed by sectional imaging when a mass lesion developed in the liver and aspiration revealed acid-fast bacilli. Case two presented with jaundice due to a hilar bile duct stricture. The patient was successfully treated by repeated bile duct stenting and later chemotherapy for tuberculosis. In both cases previous positive histology or culture would have expedited diagnosis and treatment. Acute hepatobiliary tuberculosis remains a rare disease. Suspicion of the disease and adequate biopsy are important to allow prompt appropriate treatment. PMID- 10369986 TI - Duodenal duplication cyst: MR imaging appearance. AB - The MR imaging appearance of a duodenal duplication cyst is reported. MR imaging confirmed the diagnosis suggested by ultrasound and CT scans. Fat-suppressed MR imaging before and after oral administration of the positive contrast agent Gd DTPA was able to define tissue planes between the lesion and adjacent structures, such as the head of the pancreas, providing useful information for an accurate surgical approach. To our knowledge this is the first reported case of a duodenal duplication cyst in an adult demonstrated by MR imaging. PMID- 10369987 TI - Osteomyelitis: a review of currently used imaging techniques. AB - Conventional radiographs remain the initial imaging modality involved in the diagnosis of osteomyelitis. Bone scintigraphy and its specific agents did not only eliminate the problems of inherent low sensitivity of conventional radiographs, but also increased the specificity to higher degrees. Spiral CT, on the other hand, has solved several diagnostic problems, such as osteomyelitis of the sterno-clavicular junction and hidden areas in the pelvic bones. Magnetic resonance imaging with its multiplanar capability, greater anatomic details and excellent soft tissue bone marrow contrast resolution has a significant role in surgical planning and limb preservation. Ultrasound and US-guided aspiration has recently been involved in the diagnosis and management of osteomyelitis with several advantages particularly in children. Our goal in this review is to outline the ability of various imaging techniques by comparing their strengths and weaknesses in the diagnosis of osteomyelitis. Finally, we suggest various imaging algorithms for specific clinical scenarios. Spondylitis and septic arthritis are not discussed in this review. PMID- 10369988 TI - Optimized preoperative planning of calcaneal fractures using spiral computed tomography. AB - The aim of this study was the evaluation of spiral-CT examinations in preoperative planning of calcaneal fractures supported by 3D reconstructions after electronic disarticulation. We examined 45 patients with 47 calcaneal fractures with diagnostic spiral-CT examinations in a prospective study. In addition to the conventional axial slices and sagittal reconstructions, 3D reconstructions prior to and after electronic disarticulation were performed and rated by orthopaedic surgeons and radiologists. The following diagnostic criteria were rated: involvement of articular facets, number of fragments and hindfoot deformities. Axial slices were considered to be the gold standard, because not all patients underwent surgical treatment. Axial slices showed involvement of 90 articular facets (100 %). Three-dimensional reformations after electronic disarticulation depicted 82 fractures (82 of 90, 91 %), sagittal reconstructions 63 fractures (63 of 90, 70 %). Three-dimensional reconstructions without electronic disarticulation showed five fractures (5 of 90, 5.5 %). The number of fragments was demonstrated best on sagittal reconstructions (two, three or four fragments); five fragments were diagnosed best on axial slices, and 3D reconstruction without electronic disarticulation showed only a very small number of fragments, due to overlaying bones. Hindfoot deformities (lateralisation, varus deformation, shortening) were demonstrated best on axial slices, except in terms of showing height reduction, which was demonstrated better on sagittal and 3D reconstructions. Three-dimensional reconstructions after electronic disarticulation support a clear understanding of the 3D position of the fragments and of their displacement in comparison with 3D reconstruction without electronic disarticulation, which is essential for an effective surgical reduction. Due to the potential manipulation of surface-oriented 3D reconstructions, regardless of whether electronic disarticulation is used, this imaging technique should not be considered in an isolated framework. Three-dimensional images without electronic disarticulation are superfluous in any case. PMID- 10369989 TI - Assessment of osteosarcoma response to neoadjuvant chemotherapy: comparative usefulness of dynamic gadolinium-enhanced spin-echo magnetic resonance imaging and technetium-99m skeletal angioscintigraphy. AB - The aim of this work was to study and compare the usefulness of dynamic contrast enhanced spin-echo MR imaging with high temporal resolution hydroxymethylene diphosphonate technetium-99 m skeletal angioscintigraphy in predicting the osteosarcoma histological response to neoadjuvant chemotherapy. Twelve patients with resectable osteosarcoma were prospectively monitored with dynamic MR imaging and skeletal scintigraphy before start of neoadjuvant chemotherapy, after two cycles of therapy and before surgery. Neoplasm signal intensity and activity intensity were plotted against time, and slopes were calculated for percentage increase over baseline values in the first minute. Stability and increase in slope values during or after chemotherapy were defined as a "radiological non response". Changes in slopes were compared with the "histological response" (Huvos grading). At midpoint of the chemotherapy, these two imaging modalities failed in predicting final histological response. After the completion of the chemotherapy, these imaging modalities allowed the prediction of histological response with the same accuracy (91 %). In this series, dynamic MR imaging and technetium skeletal scintigraphy provide similar results regarding the prediction of final histological response during neoadjuvant chemotherapy; these results cannot be used to modify the therapeutic protocol at midpoint of chemotherapy; these imaging tools predict accurately the histological response at the end of chemotherapy. These latter results may permit anticipation of the adjuvant chemotherapy strategy during decalcification procedures in resected osteosarcoma and thus to monitor chemotherapy in non-surgical osteosarcoma. PMID- 10369990 TI - Dysplasia epiphysealis hemimelica of the scaphoid bone. AB - We report a rare case of dysplasia epiphysealis hemimelica (DEH) in the wrist of a 7-year-old boy. Clinical, radiological and histopathological manifestations are discussed. The correct diagnosis of DEH, however, was made by the confrontation of the radiological and pathological data. The radiologist should inform the pathologist correctly about the imaging findings in order to avoid misdiagnosis of the lesion as osteochondroma. PMID- 10369991 TI - The various MRI patterns of pituitary apoplexy. AB - The aim of this study was to describe the various MRI features, in correlation to surgical and pathological findings, in patients who presented with pituitary apoplexy (PA). Eleven patients presenting with PA, were evaluated with various MR protocols including spin-echo (SE) T1-weighted sequences in 9 of 11 patients, post gadolinium SE T1-weighted sequences in only 8 of 11 patients, and with T2 weighted SE sequences in 2 of 11 patients. All patients had transsphenoidal pituitary surgery after MR studies. The severity of presenting symptoms ranged from headaches to coma. Ten patients had pituitary macroadenoma; one had a non hemorrhagic metastatic lesion into a non-adenomatous pituitary gland. Of the 11 patients, one was studied at the acute stage of PA (1 day after onset), 9 at the subacute period (3-15 days after onset), and one at the late stage (5 months after onset). Images compatible with intratumoral hemorrhage were found in all macroadenomas, whereas the metastatic pituitary lesion did not show evidence of bleeding. All gadolinium-enhanced studies showed partial tumoral enhancement. The SE T2-weighted studies demonstrated areas of low and high signal intensities in keeping with the presence of blood degradation contents. Pituitary apoplexy present with different MR features, including hemorrhagic and non-hemorrhagic characteristics on T1-weighted images. Gadolinium-enhanced images do not provide complementary diagnostic information when the presence of blood is assessed on plain images. PMID- 10369992 TI - Evaluation of the role of magnetic resonance myelography in lumbar spine imaging. AB - The purpose of our study was to evaluate the accuracy of MR myelography in depicting disc herniation in the lumbar spine when compared with conventional MRI in patients presenting with clinical evidence of disc herniation. One hundred patients referred for conventional MR imaging of the lumbar spine also had coronal MR (TR 9000 ms, TE 272 ms eff, NEX 3, echo train length 32) myelography performed. Three experienced observers compared magnetic resonance myelography (MRM) with conventional lumbar spine MR using the following variables: visibility of thecal sac and nerve roots, and the presence, location and severity of disc herniation. Disc protrusions were seen at 110 disc space levels on conventional MR images as opposed to 93 on MRM. However, only 72 % of lesions seen on conventional MR were diagnosed by MRM. Similarly, only 63.8 % of nerve root compression abnormalities seen at conventional MR were visualized when compared with conventional MRM. The sensitivity, specificity and accuracy of MRM when compared with conventional MR was 72, 93 and 85 %, respectively. The MRM technique yields images that resemble conventional myelography and may be used to help confirm abnormalities seen on conventional MR in selected cases; however, the large number of false-positive and false-negative examinations indicates that caution should be used in interpreting MRM images. PMID- 10369993 TI - Comparison of digital subtraction angiography with gadolinium-enhanced magnetic resonance angiography in the diagnosis of renal artery stenosis. AB - Renal artery stenosis (RAS) is a treatable cause of hypertension and renal failure for which no ideal screening technique is currently available. We evaluated the use of dynamic gadolinium-enhanced magnetic resonance angiography (MRA) for the diagnosis of RAS. Sixty-two patients with secondary hypertension were enrolled in the study. All patients had conventional renal angiography and gadolinium enhanced MRA. The sequence used was a 3D FMP SPGR sequence with the following parameters (TR: 26 ms, TE: 6.9 ms, flip angle 40 degrees, field of view 36 x 36 cm, matrix 246 x 256, 1 excitation). Gadolinium 0.3 mmol/kg was administered and 60 1. 5-mm-thick partitions were obtained over a duration of 3.5 min. The MRA images were then compared with conventional digital subtraction angiography (DSA) images. Conventional DSA demonstrated 138 renal arteries, whereas gadolinium-enhanced MRA demonstrated 129 (93 %). Twenty-one renal artery stenoses and four occluded arteries were seen at conventional DSA. Gadolinium enhanced MRA had a sensitivity of 88 %, specificity of 98 %, accuracy of 96 %, positive predictive value of 92 % and negative predictive value of 97 % when compared with conventional DSA. Gadolinium-enhanced MRA is an accurate technique for identifying patients with RAS. It is less sensitive in picking up accessory renal arteries. PMID- 10369994 TI - Horseshoe kidney associated with anomalous inferior vena cava. AB - Horseshoe kidney associated with anomalous inferior vena cava is a rare congenital anomaly. Radiological demonstration of this combined anomaly is also uncommon, with only two cases of preisthmic inferior vena cava with horseshoe kidney in the imaging literature. We report a case of simultaneous horseshoe kidney and inferior vena cava lying anterior to the right renal moiety diagnosed by ultrasound and computed tomography. PMID- 10369995 TI - Atypical retroperitoneal fibrosis: MRI findings. AB - A case of retroperitoneal fibrosis with an unusual perirenal involvement diagnosed at MR imaging is reported. Other conditions, such as metastatic disease or lymphoma, may be considered especially when the initial presentation is not typical. Imaging modalities in this condition are discussed. PMID- 10369996 TI - Testicular microlithiasis: US findings in six pediatric cases and literature review. AB - The aim of this article is to report on six pediatric cases of testicular microlithiasis (TM) and to review literature reports, in order to schedule US and/or other control examinations, particularly when concomitant focal or diffuse alterations of the testicular parenchymal structure are present, considering the possible association of TM with testicular tumors. Six patients (age range 4-12 years) underwent US examination for scrotal trauma (two cases) unilateral cryptorchidism (one case) follow-up after orchidopexy for bilateral cryptorchidism (one case), and varicocele (two cases). Five examinations were performed with high-frequency probes (10/13 MHz) and seven with 5/7.5-MHz frequency transducers. Follow-up US examinations were performed at different times depending on initial clinical indications, presence of underlying disease, and initial US findings. Two of the six patients underwent three US examinations, two patients underwent two US examinations, and the remaining two patients underwent only one US examination. The patients underwent a total of 12 US examinations. Microliths were bilateral in four patients and unilateral in two patients. In these two latter cases, the contralateral testis was, in one case, cryptorchid and could not be evaluated by US; in the other case it was small and hyperechogenic with orchidopexy sequelae. In three cases microliths were distributed throughout the testis. In the remaining three cases they were present in limited areas of parenchyma. As to the importance of microliths, it was defined as mild in three cases and moderate/severe in three cases. Intratubular testicular microlithiasis is a well-proved histological finding (biopsy or autopsy). More recent is the US demonstration of TM with consequent definition of its pattern: usually bilateral hyperechogenic multiple small foci without acoustic shadows with complete or partial extension to the parenchyma. Testicular microlithiasis is a rare finding. Moreover, the pediatric cases reported in the literature are very few. However, the use of high-frequency US transducers (10-13 MHz) has recently allowed an easier demonstration of this disease also in children. Of particular interest is the study of the still-debated association of microliths with other diseases such as neoplasms. Some aspects need further investigation, namely the real incidence of microliths in the healthy population, the incidence of tumors in patients with microliths, the differences between adults and children, and the different types of follow-up at different ages. In pediatric age, if TM represents an isolated sign, patients need non-invasive US follow-up until adult age. Only if TM is in association with focal lesions of testis parenchyma is it mandatory to perform biopsy or surgical treatment. PMID- 10369997 TI - Fetus-in-fetu in the scrotal sac of a newborn infant: imaging, surgical and pathological findings. AB - We report a case of fetus-in-fetu located in the scrotal sac of a newborn male infant. Plain radiography (including specimen radiography), ultrasonography and MRI clearly demonstrated vertebral column, ribs, skull, pelvic bones, femurs and a portion of tibiae and humeri. The diagnosis was confirmed by pathological examination. PMID- 10369998 TI - Coronary-subclavian steal syndrome: treatment with percutaneous transluminal angioplasty and stent placement. AB - The aim of this study was to assess the efficacy of percutaneous transluminal angioplasty (PTA) and stenting in the management of the coronary-subclavian steal syndrome (CSSS). A 56-year-old man presented with CSSS due to occlusion of the left subclavian artery. He was treated with PTA and placement of two stents in the left subclavian artery. Systolic blood pressure became equal in both arms and dizziness disappeared. There were no complications. Percutaneous transluminal angioplasty and stenting can effectively and safely manage CSSS. PMID- 10369999 TI - Percutaneous ilio-caval thrombectomy with the Amplatz device: preliminary results. AB - The Amplatz Thrombectomy Device (ATD) is a percutaneous, rotational thrombectomy catheter, capable of recirculating and homogenizing the thrombus in order to obtain mechanical clot dissolution. The authors present their experience with mechanical thrombectomy with the ATD in eight cases of ilio-caval thrombosis. Under temporary caval filter protection, the ATD was introduced through the right transjugular approach (in one patient this was used in combination with the right femoral approach) and activated for a time ranging from 90 to 180 s. Complete clearing of thrombotic material in the treated venous segments was achieved in six cases (75 %), partial success was obtained in one case (12.5 %) and failure occurred in one patient (12.5 %). One patient developed a recurrence of venous iliac thrombosis 1 week after the procedure and postphlebitic syndrome 6 months after the first episode of deep venous thrombosis, and one patient died from acute myocardial infarction, unrelated to thrombectomy session, after 3 days. A negative clinical and radiological follow-up at 3, 6, 12 and 24 months was obtained in the remaining six patients. If a fresh free-floating ilio-caval clot must be removed immediately, the ATD can be effective under temporary filter protection. PMID- 10370000 TI - Percutaneous catheter and guidewire fragmentation with local administration of recombinant tissue plasminogen activator as a treatment for massive pulmonary embolism. AB - The purpose of this article is to report four patients with massive pulmonary embolism treated with percutaneous catheter and guidewire fragmentation and local administration of recombinant tissue plasminogen activator (r-TPA). Four patients with massive pulmonary embolism initially underwent pulmonary angiography. Thrombus fragmentation was performed with both standard angiographic guidewires and catheters followed by local infusion of 41-200 mg of r-TPA. Pulmonary angiography was repeated after treatment. All patients survived with improvement in their clinical status and eventual discharge from hospital. Angiography in all patients post treatment demonstrated improvement in pulmonary perfusion (mean Miller score before treatment 22.5; mean Miller score after treatment 5.75). No patient had a significant complication. Mechanical fragmentation of the thrombus followed by local infusion of r-TPA was an effective treatment for massive pulmonary embolism in these four patients with no significant complications. PMID- 10370001 TI - Endovascular treatment of superior vena cava obstruction in patients with malignancies. AB - The aim of this study was to report our experience on the management of superior vena cava obstruction (SVCO) secondary to malignant disease, using endovascular procedures. Twenty-six patients with SVCO due to primary or secondary tumors of the lung or the mediastinum, or catheter inserted for treatment of an extra thoracic neoplasm, had an endovascular therapy which consisted of stenting, angioplasty, thrombo-aspiration or local fibrinolysis. Immediately after the procedure, rapid relief of symptoms occurred in 24 (90 %) of the patients. The mean Kishi's score decreased from 5.5 to 0.96. Immediate complications included one death related to pericarditis bleeding following fibrinolysis. Three patients relapsed after 20 days, 4 months and 6 months, and needed a second stenting. At 6 months the primary patency rate was 83 % and the secondary patency rate was 89 %. Endovascular treatment of SVCOs is a simple and safe procedure to restore the patency of the superior vena cava in malignant SVCO. It should be indicated in most cases as first-line treatment and performed as early as possible. PMID- 10370002 TI - Right diaphragmatic rupture and hepatic hernia: an indirect sign on computed tomography. AB - We report a case of blunt traumatic right diaphragm rupture with hepatic hernia. The diagnosis was first suggested by an abnormal hepatic location depicted on axial CT. This finding can be considered as a potentially new indirect sign of right diaphragm rupture in patients with blunt trauma. The diagnosis was then confirmed by reformatted CT and MR images. PMID- 10370003 TI - FDG PET-negative liver metastases of a malignant melanoma and FDG PET-positive hurthle cell tumor of the thyroid. AB - A Hurthle cell tumor (oncocytoma) of the thyroid presented as a hypermetabolic focus in a fluorodeoxyglucose positron emission tomography (FDG PET) study which was performed as staging procedure in a patient with malignant melanoma. This finding led to the initial diagnosis of a metastasis. In contrast, multiple liver metastases, seen on MRI and sonography, did not show any increased FDG uptake. Cytology results of one liver mass confirmed a melanoma metastasis, and of the neck mass, a Hurthle cell tumor. The Hurthle cell tumor was, based on clinical evidence, thought to be benign. This is the first description of a FDG PET positive benign Hurthle cell tumor, with FDG PET-negative liver metastases of a malignant melanoma, in the same patient. PMID- 10370004 TI - MR imaging: acronyms and clinical applications. AB - The intention of this article is to provide an overview of all MR imaging techniques that are accessible on most of commercially available scanners and have the potential to be used in routine clinical applications. The techniques implemented by the major vendors are briefly explained, including a comparison of the commonly used acronyms. A classification scheme is introduced which provides a reasonable illustration of similarities and differences between various techniques. The imaging techniques are divided into two main groups, the spin echo and gradient-echo sequences. Within each group is the basic sequence, those which require a preparation of the magnetization, those which use multiple echoes to fill the k-space and those which are performed in a single shot. For each technique the typical clinical applications are listed or the potential applications which have been published. PMID- 10370005 TI - Field strength and dose dependence of contrast enhancement by gadolinium-based MR contrast agents. AB - The relaxivities r1 and r2 of magnetic resonance contrast agents and the T1 relaxation time values of tissues are strongly field dependent. We present quantitative data and simulations of different gadolinium-based extracellular fluid contrast agents and the modulation of their contrast enhancement by the magnetic field to be able to answer the following questions: How are the dose and field dependences of their contrast enhancement? Is there an interrelationship between dose and field dependence? Should one increase or decrease doses at specific fields? Nuclear magnetic relaxation dispersion data were acquired for the following contrast agents: gadopentetate dimeglumine, gadoterate meglumine, gadodiamide injection, and gadoteridol injection, as well as for several normal and pathological human tissue samples. The magnetic field range stretched from 0.0002 to 4.7 T, including the entire clinical imaging range. The data acquired were then fitted with the appropriate theoretical models. The combination of the diamagnetic relaxation rates (R1 = 1/T1 and R2 = 1/T2) of tissues with the respective paramagnetic contributions of the contrast agents allowed the prediction of image contrast at any magnetic field. The results revealed a nearly identical field and dose-dependent increase of contrast enhancement induced by these contrast agents within a certain dose range. The target tissue concentration (TTC) was an important though nonlinear factor for enhancement. The currently recommended dose of 0.1 mmol/kg body weight seems to be a compromise close to the lower limits of diagnostically sufficient contrast enhancement for clinical imaging at all field strengths. At low field contrast enhancement might be insufficient. Adjustment of dose or concentration, or a new class of contrast agents with optimized relaxivity, would be a valuable contribution to a better diagnostic yield of contrast enhancement at all fields. PMID- 10370006 TI - Primary aorto-duodenal fistula: evaluation with computed tomography. PMID- 10370007 TI - Persistent trigeminal artery associated with basilar artery hypoplasia: MR and MRA findings. PMID- 10370008 TI - Association between spinal extradural and cerebral arteriovenous malformations appearing as acute paraplegia after epidural hemorrhage. PMID- 10370009 TI - Results of spiral CT in patients with fractures of the craniocervical junction suspected on conventional radiographs. PMID- 10370010 TI - Quiz of the month: foreign body perforation of the cervical esophagus caused by a walnut. PMID- 10370011 TI - The effect of phospholipids and mucin on bacterial internalization in an enterocyte-cell culture model. AB - A primary component of the intestinal mucous layer that functions as a barrier to luminal bacteria is mucin, a high-molecular-weight glucoprotein. In addition, the mucous layer also contains other important elements such as phospholipids (PLs), which may effect bacterial translocation (BTL). It has been reported that mucin inhibits Escherichia coli translocation; however, the effect of PLs on intestinal permeability is still controversial. We have recently reported that the concentration of mucous PLs is higher in neonatal as compared to adult rabbits. The functional significance of these biochemical differences on BTL remains to be determined. The aim of this study was to evaluate the effect of PL and mucin composition on BTL in a human enterocyte-cell culture model. Human enterocyte Caco-2 cells were seeded in 24-well tissue-culture plates and grown for 14 days to confluence. The monolayers were pretreated with phosphate buffered saline as control, 10 mg/ml or 20 mg/ml mucin, 0. 5 mM or 1.0 mM PL mixture based on neonatal (NPL) and adult (APL) composition, and 10 mg/ml mucin with 0.5 mM either APL or NPL mixtures 30 min before a 120-min incubation period at 37 degrees C with 10(8) colony forming units (CFU) of E. coli C25. Non-internalized bacteria were killed by the addition of gentamicin. Internalized bacteria were quantified by counting CFU from enterocyte-cell lysates grown on MacConkey's agar. Mucin inhibited bacterial internalization, while both compositions of PLs promoted it. Mucin added to the PL solution also diminished the stimulatory effect of PLs on bacterial internalization. These results indicate that the increased concentration of PLs found in the intestinal mucous layer of neonates, and/or the alteration in the balance between PLs and mucin, may play a role in the increased BTL seen in neonates. PMID- 10370012 TI - The effect of mucin on bacterial translocation in I-407 fetal and Caco-2 adult enterocyte cultured cell lines. AB - Although the intestinal mucosa forms a crucial barrier between the host and the environment, bacterial translocation (BT) occurs frequently in neonates and may be a source of sepsis. The intestinal mucous gel layer is thought to be a vital component of the gut barrier and is composed, in part, of a family of glycoproteins known as mucins. Our aim was to study the effects of mucin on BT in an enterocyte cell-culture model using a fetal (I-407) and an adult (Caco-2) intestinal cell line. I-407 and Caco-2 cells were grown to confluence on porous filters in a two-chamber Transwell system. The integrity of the monolayers was confirmed by transepithelial electrical resistance (TEER) and permeability using the macromolecule dextran blue. Cells were treated with mucin (40 mg/ml) prior to inoculation of 1 x 10(6) Escherichia coli C25. The magnitude of BT was determined quantitatively by culturing the samples from the basal chamber of the wells and was expressed as log 10 [Colony Forming Units (CFU)/ml]. Statistical analysis was performed by the Mann-Whitney U test with statistical significance at P < 0.05. Mucin inhibited BT across both fetal and adult cultured enterocyte monolayers; however, the inhibitory effect was less on the fetal cells compared to the adult cells. Dextran-blue studies showed that monolayers were intact throughout the experiments. Despite 98% inhibition of BT, mucin had a statistically significant effect on post-bacterial inoculation TEER in Caco-2 cells and no effect in I-407 cells. The ability of mucin, a mucous-barrier glycoprotein, to inhibit BT across immature intestinal enterocytes, as in the neonate, may be diminished compared to mature adult enterocytes. PMID- 10370013 TI - Effect of growth hormone on bacterial translocation in experimental short-bowel syndrome. AB - Despite the adaptive process triggered in the remaining intestine by massive bowel resection, bacterial translocation (BT) is frequent under these conditions. Several trophic factors, including growth hormone (GH), are involved in the process of adaptation in short-bowel syndrome (SBS). However, the effect of GH on BT has not been investigated experimentally. The aim of this study was to test the hypothesis that GH administration prevents BT in rats with SBS receiving only parental nutrition (PN). Nineteen adult Wistar rats underwent central venous cannulation and were randomly assigned to one of two groups receiving for 10 days two treatment regimes: PN group (n = 10): fasting, all-in-one PN solution (300 ml. kg. 24 h, 280 kcal/kg. 24 h), 80% gut section including ileocecal valve; GH group (n = 9): fasting, same PN regime and resection plus GH 1 mg/kg s.c). At the end of the experiment, the rats were killed and mesenteric lymph nodes (MLN) and samples of systemic and portal blood were obtained and cultured. Several samples of full-thickness jejunal wall were taken for determining cell proliferation index (PCNA) and mucosal thickness. Jejunal mucosal thickness increased by 30% and PCNA index by 35% in GH-treated rats in comparison with those treated with PN alone. However, contrary to our expectations, BT expressed by positive culture of intestinal flora in portal blood, MLN, or systemic blood was found in 60% of PN and 87% of GH animals (P = 0.1). Translocation to the general circulation expressed by the presence of organisms in systemic blood was detected in 0% of PN and 44% of GH rats (P < 0.05). Although exogenous GH improves gut mucosal structure in rats with SBS treated with PN, it seems to increase rather than decrease mucosal permeability to intestinal bacteria. PMID- 10370014 TI - Long-term insulin-secretory function of islets of Langerhans encapsulated with a layer of confluent chondrocytes for immunoisolation. AB - Islet transplantation is a potential cure for diabetes mellitus. The major problem for broad clinical application remains the prevention of transplant rejection without major side effects. Immunoisolation is an experimental strategy to prevent rejection by separating the transplanted cells from the host immune system using a barrier device. Current methods use artificial, not completely inert materials as barrier devices and induce an unwanted foreign-body (FB) reaction. Using the recipients of own cells for encapsulation, the FB reaction could be prevented. This study describes a new method of encapsulation of islets of Langerhans within a capsule of chondrocytes, which may serve as an immunoisolation barrier utilizing the immunoprivileged properties of the chrondrocyte matrix, and demonstrates the functional survival of the encapsulated islets in vitro. PMID- 10370015 TI - Heterotopic hepatocyte transplantation utilizing pancreatic islet cotransplantation for hepatotrophic stimulation: morphologic and morphometric evaluation. AB - Hepatocyte transplantation using three-dimensional (3D) matrices is under evaluation as an alternative therapy for liver diseases. It is known that hepatotropic stimulation optimizes hepatocyte engraftment. We investigated hepatotrophic stimulation by portocaval shunt operation (PCS) or pancreatic islet cotransplantation (ICT) over a period of 6 months. Lewis rats served as donors and recipients, respectively. One week prior to hepatocyte implantation PCS (group A) or a sham operation (groups B-D) was performed. Four polyvinyl-alcohol matrices, each containing 1.25 x 10(7) hepatocytes (groups A and C), 1.25 x 10(7) hepatocytes and 500 islets (group B), or cell-free culture medium (group D, control) were implanted between recipients' small-bowel mesenteric leaves. One, 3, and 6 months after implantation eight polymers from each group were harvested and analyzed by morphometry, PAS reaction, and immunohistochemistry for insulin, glucagon, and bromodesoxyuridine. Morphologically healthy-appearing hepatocytes were found in all cell transplantation groups at all times. Stimulation by either PCS or ICT significantly increased hepatocyte area at 1 and 6 months compared to unstimulated specimens (group C). Over time, an increase in hepatocyte area was noted in all groups. There were no significant differences in proliferation ratios between the three experimental groups. The initially reduced PAS reaction became normal after 3 months. 3D matrices provided a sufficient environment for transplanted hepatocytes and islets. Hepatocytes proliferated and maintained differentiation independent of hepatotrophic stimulation for at least 6 months when 3D matrices were utilized. ICT efficiently stimulated transplanted hepatocytes by means of hepatocyte area. These results justify further research on hepatocyte transplantation and ICT with regard to clinical application. PMID- 10370016 TI - Effect of antenatal glucocorticoid administration on insulin-like growth factor I and II levels in hypoplastic lung in nitrofen-induced congenital diaphragmatic hernia in rats. AB - There is increasing evidence to suggest that insulin-like growth factors (IGF) I and II play a crucial role in fetal lung development. Expression of IGF-I and II has been demonstrated to be predominant during fetal life and decreases prior to birth. Antenatal glucocorticoids are reported to improve lung immaturity. The aim of this study was to investigate the effect of antenatal glucocorticoid administration on IGF-I and II expression in nitrofen-induced congenital diaphragmatic hernia (CDH) in rats. A CDH model was induced in pregnant rats following administration of 100 mg nitrofen on day 9.5 of gestation (term = 22 days). Dexamethasone (0.25 mg/kg) was given intraperitoneally on days 18.5 and 19.5 of gestation. Cesarean section was performed on day 21. The fetuses were divided into three groups: I, normal controls; II, nitrofen-induced CDH; and III, nitrogen-induced CDH with antenatal dexamethasone treatment. mRNA was extracted from whole lung and a reverse transcription-polymerase chain reaction (RT-PCR) was performed to evaluate the relative amounts of IGF I and II mRNA. Levels of mRNA were expressed as a ratio of the band density divided by that of beta-actin, a housekeeping gene known to be expressed at a constant level. Immunohistochemistry using anti-rat IGF I and II antibody was also performed in each group. Levels of IGF I mRNA were significantly increased in group II (0.50 +/- 0.08) compared to group I (0.34 +/- 0.10) or group III (0.32 +/- 0.06) (P < 0.05). Levels of IGF II mRNA were also significantly increased in group II (0.95 +/- 0.20) compared to group I (0.42 +/- 0.07) or group III (0. 31 +/- 0.09) (P < 0.05). Strong IGF I and II expression was observed in the hypoplastic CDH lung (group II), mainly in the bronchiolar epithelium. IGF I and II expression in group I and III lungs was either absent or weak. The finding of significant reductions in IGF I and II mRNA and protein levels in dexamethasone-treated CDH lung suggest that dexamethasone may accelerate the fetal stage of lung development. PMID- 10370017 TI - Pulmonary neuroendocrine cells in nitrofen-induced diaphragmatic hernia and the effect of prenatal glucocorticoids. AB - The high mortality associated with congenital diaphragmatic hernia (CDH) is due to pulmonary hypoplasia and hypertension, structural and functional abnormalities which can to some extent be ameliorated by prenatal administration of glucocorticoids. In the hypoplastic, hypertensive lungs of neonatal rats in which CDH has been induced by nitrofen, those pulmonary neuroendocrine cells (PNCs) containing calcitonin gene-related peptide (CGRP) increase in number, and it has been suggested that this might be due to inhibition of secretion of the peptide, the consequent decrease in its vasodilatory effects contributing to the hypertension. Whether this increase affects the entire population of PNCs, however, and how these cells are affected by administration of prenatal glucocorticoids, is unknown. As revealed by immunolabelling for protein gene product (PGP) 9.5, a general marker of NCs and expressed per cm2 tissue section, the total PNC population in rats with nitrofen-induced CDH was significantly greater than in controls receiving only olive oil (672 vs 375/cm2, P = 0.03) and was further increased (824 per cm2) in animals treated prenatally with dexamethasone (n = 8 in all groups). The increase in the total PNC population in rats with CDH is similar in magnitude to that described for the CGRP-containing subpopulation. Since the major role of the products of PNCs is now thought to be the regulation of development of pulmonary tissues and their response to injury, it is probable that the expansion of their population in the abnormal lungs associated with CDH is an adaptive response to pulmonary maldevelopment, a response possibly augmented by exogenous corticosteroids. PMID- 10370018 TI - The tracheobronchial tree is abnormal in experimental congenital diaphragmatic hernia. AB - Neonates with congenital diaphragmatic hernia (CDH) have other malformations that contribute to the high mortality. The nitrofen rat model allows experimental study of these anomalies. This study examines whether the tracheobronchial tree is also abnormal in this model. Time-mated rats received 100 mg nitrofen on gestational day 9. 5; 90 fetuses were harvested on day 21 (near full term) and dissected. The trachea and bronchi were stained with alcian blue-alizarin red and their anatomy was examined by transillumination under a microscope. The findings were compared with those of 11 suitable controls. Control pups had no malformations. Those with CDH (n = 57) had significantly decreased numbers of tracheal rings in comparison with controls (22.9 +/- 1.9 vs 26 +/- 1.9, P < 0.05) and 40/57 had fragmented rings (0 in controls). Twelve CDH animals had, in addition, tracheal stenoses of variable severity, sometimes related to vascular rings. Nitrofen fetuses without CDH (n = 33) had only short tracheas and 4 had mild stenoses. Nitrofen-exposed fetuses have, in addition to lung hypoplasia and sometimes CDH, severe tracheobronchial anomalies that suggest the involvement of pathogenetic mechanisms capable of acting on various tissue components. The genetic control of organogenesis is most probably disturbed by the teratogen. PMID- 10370019 TI - Lung hypoplasia caused by nitrofen is mediated by down-regulation of thyroid transcription factor TTF-1. AB - Prenatal exposure to nitrofen induces lung hypoplasia and diaphragmatic hernias very similar to those in human disease, but the mechanisms are still unknown. Thyroid transcription factor 1 (TTF-1) is involved in lung ontogeny and regulation of the expression of surfactant proteins, and is likely abnormally expressed in nitrofen-induced lung hypoplasia. This study examines the effect of nitrofen on TTF-1 messenger RNA (mRNA) expression in the lungs of prenatal rat fetuses and a human lung-cell line (NCI-H441) that expresses both TTF-1 and surfactant proteins in vivo. Lungs from preterm fetuses harvested from rats with 100 mg nitrofen on gestational day 9.5 and NCI-H441 cells maintained in RPMI medium containing 10% fetal bovine serum and exposed to nitrofen for different times and concentrations were assayed for TTF-1 mRNA by northern blot analysis. mRNA for TTF-1 was decreased in nitrofen-exposed pups in comparison with controls, and exposure to nitrofen caused a dose- and time-related decrease in TTF-1 expression in H441 cell cultures. These results indicate that nitrofen downregulates TTF-1 both in vivo and in vitro. Since this interferes with lung development, it is reasonable to accept that lung hypoplasia in this model is in part due to the direct effect of the teratogen rather than to compression by the abdominal viscera herniated into the thorax. This mechanism should be explored in the clinical setting. PMID- 10370020 TI - Altered intramuscular innervation and synapse formation in internal sphincter achalasia. AB - Internal anal sphincter achalasia (IASA) is a condition with a clinical presentation similar to Hirschsprung's disease, but with the presence of ganglion cells on rectal biopsy. The diagnosis of IASA is made on anorectal manometry, which demonstrates the absence of a rectosphincteric reflux on rectal balloon inflation. In order to understand the nature of neuronal abnormalities in this condition, we performed immunohistochemistry using PGP 9.5 (a general neuronal marker) and synapsin I (a presynaptic marker) in IAS specimens from 10 patients with IASA and 8 normal controls. In the IAS of normal controls, there were many PGP 9.5 and synapsin I-positive nerve fibers. In IASA PGP 9.5-immunoreactive fibers were markedly reduced and synapsin I-positive fibers were either absent or markedly reduced. Our findings demonstrate that the IAS in achalasia patients has defective intramuscular innervation as well as defective innervation of the neuromuscular junction, thereby contributing to the motility dysfunction. PMID- 10370021 TI - Whole-mount NADPH-diaphorase histochemistry is a reliable technique for the intraoperative evaluation of extent of aganglionosis. AB - Multiple seromuscular biopsies at three levels (narrow segment, transitional zone, and dilated segment) were taken and investigated intraoperatively to determine the extent of aganglionosis. Using the whole-mount preparation technique, circular muscle fibers were separated from the specimens. After a short prefixation, the muscle fibers were stained by the NADPH-diaphorase technique and were examined within 20-25 min. A fine and dense neuronal meshwork was observed between circular muscle fibers in the normal and ganglionic part of the bowel. In contrast, there was a complete lack of NADPH-diaphorase-positive fibers in the circular muscle of aganglionic colon. In the transitional zone, NADPH-diaphorase-positive fibers were markedly reduced compared to the ganglionic region. The density of these fibers increased and attained normal levels in the proximal bowel above the transition zone. These results suggest that whole-mount NADPH-diaphorase histochemistry is a three-dimensional technique suitable for the intraoperative evaluation of extend of aganglionosis. The technique is sufficiently rapid to be used in conjunction with routine frozen sections to assist in the diagnosis and in selecting the optimal level of resection at the time of pull-through operation. PMID- 10370022 TI - Increased expression of transforming growth factor-alpha in infantile hypertrophic pyloric stenosis. AB - Infantile hypertrophic pyloric stenosis (IHPS) is characterized by hypertrophy of the pyloric muscle. The growth of smooth-muscle cells (SMCs) is regulated by several growth factors. Transforming growth factor-alpha (TGF-alpha) is a growth regulatory peptide found in a wide range of embryonic and adult tissues. It has been recognized that TGF-alpha has growth-promoting effects in vascular and visceral SMCs. The aim of this study was to investigate whether TGF-alpha plays a role in the pyloric-muscle hypertrophy in IHPS. Full-thickness pyloric-muscle biopsy specimens were obtained at the time of pyloromyotomy from 10 IHPS patients (age range 24-76 days). Age-matched control material included 10 pyloric-muscle specimens taken at autopsy in patients without evidence of gastrointestinal disease. Indirect immunohistochemistry was performed using the ABC method with anti-TGF-alpha polyclonal antibody. In-situ hybridization was performed using a digoxigenin-labelled, TGF-alpha-specific oligonucleotide probe. There was a marked increase in TGF-alpha immunoreactivity and messenger RNA (mRNA) expression in SMCs in pyloric circular and longitudinal muscle in IHPS specimens compared to controls. The increased expression of TGF-alpha mRNA together with increased TGF alpha immunoreactivity in IHPS suggests increased local synthesis of TGF-alpha by pyloric SMCs, causing pyloric-muscle hypertrophy. PMID- 10370023 TI - Notochord involvement in experimental esophageal atresia. AB - Esophageal atresia (EA) is often accompanied by vertebral defects and other anomalies. The adriamycin rat model of EA has disclosed the embryology of the malformation and shown that the vertebrae and notochord are also abnormal. This study describes the nature of notochord malformations in rat embryos exposed to adriamycin. Time-mated rats received either 1.75 mg/kg adriamycin or vehicle i.p. on gestational days (E) 6 to 9; E-12, E-12.5, and E-13 embryos were harvested, embedded in paraffin, and serially sectioned at 3 microm in transverse plane from the head to the stomach for subsequent PAS staining. The findings in both groups were compared at the three endpoints. Control embryos had neither tracheoesophageal nor notochord malformations. On day 12, only 11/36 adriamycin embryos were normal; 7/36 had abnormal notochords, 11/36 had EA, and 7/36 had both. The corresponding figures for 12.5 days were 12/27, 0/27, 7/27, and 8/27 and those for the day 13 7/23, 5/23, 3/23, and 8/23. The malformed notochords were thickened, bifurcated, or trifurcated in the sagittal plane. The simultaneous presence of notochord and esophageal malformations suggests a direct link between both defects, but our observation of isolated occurrence of both shows that they reflect two expressions of the profound disturbance of embryonic para-axial organization responsible for the cluster of malformations rather than a cause-effect association. PMID- 10370024 TI - Unilateral agenesis of the diaphragm: a separate entity or an extremely large defect? AB - Since the mid-1980s, unilateral agenesis of the diaphragm (DA) has attracted the attention of paediatric surgeons as more babies affected by this extreme form of congenital diaphragmatic hernia (CDH) survive. Some authors believe that it represents a separate clinical entity. We undertook a retrospective analysis of all babies with CDH treated in the South-West Regional Paediatric Surgical Centre in Bristol between 1981 and 1995. Of 108 babies 16 (14.8%) were identified as having DA. All presented with severe respiratory distress from birth. In comparison to the group of patients with postero-lateral hernia, neonates with DA had lower Apgar scores and required longer preoperative stabilisation with inotropic support and vasodilators. Nine were subjected to operation and all required diaphragmatic replacement. Only 3 survived; thus, mortality in the DA group was 81.25%, and among those who underwent surgery 66.6% The same data for babies with postero-lateral hernia were 15.2% and 7.2%, respectively. Our results indicate that DA is associated with high morbidity and mortality, but we have not found any evidence that this anomaly is a distinct entity. In addition, we reviewed all post-mortem reports of fetuses with diaphragmatic defects available for the same period. Of 19 fetuses, 10 (52.6%) had DA. The morphological details of the diaphragmatic defect and the presence of associated anomalies were analysed. Our observations support the hypothesis that DA occurs in the very early stages of embryonic life and may be attributed to developmental arrest of the septum transversum. PMID- 10370025 TI - Helicobacter pylori infection in postoperative patients with biliary atresia. A benign colonization? AB - The prevalence of Helicobacter pylori infection in postoperative patients with biliary atresia (BA) was investigated in relation to esophageal varices, portal hypertensive gastropathy (PHG), and peptic ulcer disease (PUD) in 25 Japanese patients (10 boys and 15 girls) aged from 16 months to 20 years. Gastric biopsy specimens obtained during endoscopy were used for both the rapid urease test and modified Giemsa staining. The patients were classified into three groups according to liver function: 15 in group A (total bilirubin [TB] < 1.0 mg/dl), 7 in group B (1.0 /= 2.0 mg/dl). Esophageal varices were found in 19 patients (60% of group A and all patients in groups B and C) and PHG in 3 group B patients. However, no gastric or duodenal ulcers were found in any case. Only 2 patients (8%) had H. pylori colonization of the gastric mucosa. Both, however, belonged to group A, and the degree of chronic neutrophilic infiltration of the mucosal layer was as mild as that of the other patients. The prevalence of H. pylori infection and PUD in postoperative patients with BA was quite low, and a pathological relationship with the severity of liver disease could not be determined in these patients. PMID- 10370026 TI - Unsuccessful air-enema reduction of intussusception: is a second attempt worthwhile? AB - Pneumatic reduction of idiopathic intussusception is successful in about 80% of cases, while 60% of the failures are reduced at surgery without resection. To determine whether delayed, repeated attempts at enema reduction of failures would reduce the need for operation in selected cases, over a 2-year period (1994-1996 inclusive), 17 infants with idiopathic intussusception underwent delayed repeat enemas 2-19 h following the first failed attempt at reduction. Clinical parameters and radiologic findings were evaluated with respect to outcome. Ten intussusceptions were successfully reduced after the second attempt in 9 and after the fourth attempt in 1. Seven children underwent a laparotomy, 5 because of failure of progressive reduction at air enema (AE). Two were taken to surgery early in the series, 1 because of perforation during a second attempt and 1 while awaiting a third reduction attempt. The 10 successful reductions all showed progressive movement of the intussusceptum on each AE; the 2 who perforated failed to show progressive reduction on their second AE. Because of these cases, the remaining 5 were referred to surgery because of failure of progressive reduction of the intussusceptum on the second attempt. At laparotomy, of the 7 unsuccessful reductions, 4 required resection and 3 had difficult manual reduction. The presence of vomiting, a mass, and/or bloody stools were not predictors of outcome. Failures had higher body temperatures (38.1 +/- 0.3 vs 37.4 +/- 0.1 degrees C, P = 0.07), heart rates (153.7 +/- 8 vs 136.9 +/- 2.1 min, P = 0.03), and longer duration of symptoms (36.8 +/- 4 vs 21.3 +/- 3.6 h; P = 0.01) than successes. Delayed repeat AEs may be safe and effective in selected cases of idiopathic intussusception, but should be considered only if significant movement of the intussusceptum is noted at each attempt. The ideal time for repeat AE reduction prior to surgery is not established, but 2-4 h appears appropriate. Pyrexia, tachycardia, and duration of symptoms greater than 36 h are relative contraindications to this course of management. PMID- 10370027 TI - Hydatid disease of the liver in childhood. AB - The records of 100 children with hydatid disease were reviewed retrospectively from 1978 to 1997; 43 were girls and 57 were boys. The mean age was 9.14 years; 61 patients had 124 hepatic cysts. Presenting symptoms were asymptomatic abdominal masses, found masses incidentally during ultrasonography (US), or acute abdomen. Plain X-ray films, US, or computerized tomography (CT) are sufficient for diagnostic evaluation in endemic areas. In the differential diagnosis, laboratory investigations such as the Casoni and Weinberg tests, indirect hemagglutination, eosinophilia, and ELISA were also used. These tests may give negative results, however, in some patients with hydatid disease. The mean follow up time was 10.5 years (range 1-18 years), the mean duration of hospitalization 7 days. The complication rate was 3.6%. Mortality was 3.27% and occurred after the administration of formaldehyde and hypertonic scolicidal agents. Hydatid disease of the liver can be treated medically in selected patients; conservative surgical approaches that save as much parenchyma as possible, such as partial cystectomy and capitonnage, are indicated in the other cases. PMID- 10370028 TI - Anal transposition without colostomy: functional results and complications. AB - Rectovestibular fistula (RVF) is the most common form of anorectal anomaly in female infants. In the surgical repair of these malformations, most pediatric surgeons use cutback, fistula transposition with or without colostomy, and lately, posterior anorectoplasty with colostomy. This is a retrospective evaluation of the functional results and complications in 47 patients who underwent fistula transposition without colostomy for the treatment of a RVF. We prefer to perform the operation when the rectovaginal septum is amenable to dissection (width >2 mm). All patients had voluntary bowel movements; 28 (60%) had completely normal bowel habits, 45 (96%) good and only 2 (4%) fair. We did not encounter serious surgical complications such as infection dehiscence, and fistula recurrence. We thus prefer anal transposition without colostomy to other modes of surgical therapy for RVF. PMID- 10370029 TI - Resection of foregut-derived duplications by minimal-access surgery. AB - Eight children underwent minimal-access surgery (MAS) for duplications of foregut derivatives. The efficacy and safety of this approach are reviewed. The seven patients with mediastinal lesions had video-assisted thoracoscopic resection. One lesion presented as a subdiaphragmatic esophageal diverticulum, which was excised laparoscopically. Between March 1991 and October 1997, eight children were treated. Mean age was 27 months and mean weight was 11. 4 kg. Mean operating time was 106 min, and mean postoperative hospital stay was 4.5 days (median = 2 days). Persistent air leaks occurred in two patients who had centrally-located bronchogenic cysts. One of these, who had undergone subtotal excision with laser photoablation of the remaining cyst mucosa, developed a recurrence that was excised at thoracotomy. We conclude that esophageal and bronchogenic cysts and duplications may be safely excised by MAS in children, with excellent cosmetic and functional outcome. Two technical points are noted: (1) a thoracostomy tube is required for central mediastinal lesions; and (2) complete excision is required to prevent recurrence. PMID- 10370030 TI - Perineal-mound defects. AB - Perineal-mound (PMD) and genital-fold defects cause anorectal malformations (ARM) in both sexes. They are common in females and usually present as a low anomaly (except rectovestibular fistula). They are rare in males and present as an intermediate anomaly. A common embryological explanation for these defects with varied presentation in males and females is discussed. These anomalies should be grouped separately in the classification of ARM. We present five male patients with PMD, three of whom had imperforate anus with rectobulbar fistula and perineal hypospadias and two who had imperforate anus with rectoperineal fistula. PMID- 10370031 TI - Anderson-Hynes pyeloplasty in horseshoe kidney in children: is it effective without symphysiotomy? AB - Contemporary reports on surgery for horseshoe kidney (HK) still recommend isthmotomy and lateropexy to complete an open pyeloplasty. To evaluate whether simple Anderson-Hynes pyeloplasty without symphysiotomy is effective for relief of ureteropelvic junction obstruction (UPJO) in HK, we studied the records of ten children, two of whom had bilateral UPJO. Only one child presented with calculi; 11 units were operated upon for UPJO, 1 needed a partial nephrectomy. The surgical outcome was evaluated with emphasis on the changes in renal drainage and function assessed by ultrasonography and diuretic renal scans. Associated vesicoureteral reflux was observed more often (25%) than with UPJO in normal kidneys. Obstruction was caused by a crossing lower-pole vessel in three cases, a high ureteral insertion in two and narrowing of the UPJ 7. Postoperative follow up (mean 5.5 years) revealed improved renal function and good drainage in all cases. Hydronephrosis vanished in 7, whereas grade 2 hydronephrosis remained in two children with former refluxive megaureter and grade 3 in one. All children are doing well and have no symptoms due to the persistent isthmus (Rovsing syndrome). It is concluded that simple Anderson-Hynes pyeloplasty via a flank incision is a highly effective and safe procedure for treating UPJO in HK. PMID- 10370032 TI - Testicular catch-up growth after varicocele correction in adolescents. AB - We evaluated retrospectively the outcome of artery-sparing (AS) versus non-artery sparing (NAS) laparoscopic varicocelectomy and measured any reversal of testicular growth. Twenty patients (13 left and 7 bilateral varicoceles) were evaluated after surgery. A total of 27 varicocelectomies (20 AS and 7 NAS) were performed. The indication for surgery was smaller testicular size on the affected side in all patients and discomfort/pain in 3. The mean age was 12.9 years (range 8-15 years) at surgery. The testicular volumes were determined clinically and by color Doppler sonography (US). The follow-up time was 6-48 months after surgery. There were 4 recurrences out of 27 varicocelectomies (15%), of which 1 has been reoperated. Testicular volumes were equal in both groups after surgery, indicating catch-up growth except in the cases with minor recurrences (2 AS and 2 NAS varicocelectomies). In 12 testes, dilated veins in the pampiniform plexus were revealed by US. No severe intraoperative complications occurred. Three patients had a hydrocele after surgery (11%). These data show that there is testicular catch-up growth after varicocelectomy, but some questions remain unanswered: (1) should the remaining dilated veins detected by Doppler US be tackled; and (2) is an AS operation worthwhile? PMID- 10370033 TI - T-lymphocyte subsets in the contralateral testis after unilateral blunt testicular trauma in pre-pubertal mice. AB - Although spermatozoa express antigens, they normally do not produce an immunological response because of the blood-testis barrier and the predominance of CD8(+) T-lymphocytes in the rete testis. Unilateral blunt testicular trauma (UBTT) has been reported to decrease fertility. The present study was designed to evaluate the sub-populations of T-lymphocytes in mice with testicular trauma. Twenty male mice aged 20 days were randomized into control and test groups. At about 70 days of age the contralateral testis was harvested, cell suspensions were prepared, and immunofluorescence staining was performed for detection of CD4(+ )and CD8(+) T-lymphocytes by flow-cytometry. The ratio of CD8(+) and CD4(+) lymphocytes was significantly higher (P < 0.001) in the control mice compared to the UBTT group (1.3 +/- 0.3 vs 0.5 +/- 0.01). The results suggest that UBTT alters the CD8(+)/CD4(+) ratio in the contralateral testis, which may have an important bearing on the pathogenesis of infertility in cases of testicular injury. PMID- 10370034 TI - Urethral mobilization and meatal advancement: a surgical principle in hypospadias repair. AB - A technique of urethral mobilization and advancement in hypospadias repair using the urethral elasticity to partially or completely bridge the defect in urethral length was employed in 56 children. In 46 with distal hypospadias it was the only procedure used. In 10 with proximal hypospadias, it was combined with other techniques. In distal hypospadias, no postoperative fistula occurred. Complications of the operation were 3 meatal stenoses that responded to dilatation, 1 urethral injury immediately repaired with no consequent fistula, and 1 chordee that was subsequently corrected. Of the 10 children with proximal hypospadias, 3 developed minor fistulae and 1 meatal stenosis. Urethral mobilization was found to be a safe and effective procedure in the management of hypospadias. It could be the only procedure required in distal hypospadias, or in combination with other procedures in proximal hypospadias. PMID- 10370035 TI - Apoptosis and bcl-2 oncogene expression in Wilms' tumor. AB - Wilms' tumor (WT) usually has a good outcome, although a poor prognosis is often related to more advanced stages and anaplastic features. Apoptosis occurs with variable frequency in malignant tumors, and may have a role in reducing their growth rate. The bcl-2 proto-oncogene inhibits apoptosis, and the consequent increase in the number of cells may play a role in the development of tumors. The aim of this study was to analyze the role of apoptosis and bcl-2 expression in WT. Twenty-six resected WT specimens were studied; 12 patients had stage I tumor, 4 stage II, 5 stage III, 3 stage IV, and 2 stage V. Twenty-three tumors were classified as favorable histology (FH) and 3 as unfavorable (UH). The mean follow up was 34 months; 22 patients were alive and 4 were dead (2 with FH: 1 stage III and 1 stage IV, and 2 with UH stages 4). Apoptosis was detected by the in-situ end-labelling technique; bcl-2 expression was detected by immunohistochemistry. An apoptotic index (AI) was calculated as the ratio of apoptotic to normal cells in each specimen. The AI was lower in higher tumor stages, with a significant difference between stages I and IV (P < 0.05). In cases with UH, Al was lower than in tumors with FH (P < 0.01). The AI was also lower in patients who died than in those who survived (P < 0. 01). In all specimens no correlation between AI and bcl-2 expression was observed. Progression to advanced stages of WT and a poor prognosis f anaplastic tumors may be linked with disruption of the mechanisms that control apoptosis. Bcl-2 does not play a role as a regulator of apoptosis in WT, other oncogenes and tumor-suppression genes may be more involved in inhibiting apoptosis in WT. PMID- 10370036 TI - Accessory limbs associated with spina bifida - a second look. AB - An accessory limb associated with spina bifida was already reported by the authors. We had then described it as a result of a very early splitting of the limb bud arising from the paraxial mesoderm. We have subsequently seen three other such cases, which are described in this report as well as a review of five other cases in the literature. It is proposed that the growth of the accessory limb occurs from a mesodermal blastema that is a result of de-differentiation from Schwann cells. PMID- 10370037 TI - Sonographic management of infantile clavicular fractures. AB - A new method for radiation-free management of infantile clavicular fractures is presented. Forty-nine infants with clavicular fractures were examined radiologically and sonographically by independent examiners. The two imaging techniques were compared with regard to practicability, diagnostic results, and assessment of the healing process. The paired t-test showed no significant difference between X-ray and ultrasound (US) for the criteria of practicability and primary diagnostic results. With regard to assessment of the healing process, a highly significant advantage in favor of US was registered. One week before radiographs revealed any sign of bone healing, stress-resistant, stable healing of tissue is seen on US. Even consolidation disorders such as early signs of pseudarthrosis are detected very early by US. This enormous diagnostic advantage reduces the duration of immobilization, the frequency and number of follow-up examinations, and last but not least, provides a cost-effective method of treatment without exposure to ionizing radiation. PMID- 10370038 TI - Hyaluronate, tetrachlorodecaoxide, and galactolipid prevent adhesions after implantation of Gore-Tex and dura mater into the abdominal wall in rats. AB - Gore-Tex (GT) and dura mater (DM) are used as prosthetic materials for the closure of the abdominal wall defects, however, they create intra-abdominal adhesions. This study addresses the question of which substances can reduce these adhesions. In rats, Gore-Tex and DM were placed on the inner abdominal wall. Two weeks later the animals were killed; the anterior abdominal wall was excised and photographed. The photographs were digitized and the surface area covered by adhesion was measured by computer analysis. In animals where DM or GT was implanted without the addition of an anti-adhesive substance, 45% of the DM and 34% of the GT surface was covered by adhesions. When hyaluronate (HA), tetrachlorodecaoxide, or galactolipid was applied to the bowel intra-operatively, adhesions were found on only 14%, 11% or 8% of the GT surface. For DM, only HA was effective, and reduced adhesions to 9% and 10%, respectively. Plasmin, taurolidine, and streptokinase-streptodomase were ineffective in preventing adhesions in both DM and GT. PMID- 10370039 TI - Congenital cystic adenomatoid malformation of the lung associated with esophageal atresia and tracheoesophageal fistula. AB - Bronchopulmonary malformations associated with esophageal atresia (EA) and tracheoesophageal fistula (TEF) are extremely rare. The authors describe a case of type II congenital cystic adenomatoid malformation (CCAM) of the right lower lobe associated with EA and TEF (Vogt-Gross type C) in a full-term female infant. The CCAM presented as an incidental radiologic finding, and a contralateral tension pneumothorax developed shortly after surgical repair of the EA. Early recognition of this rare association is essential for correct operative management. PMID- 10370040 TI - Pyloric atresia associated with epidermolysis bullosa, malrotation, and high anorectal malformation with recto-urethral fistula: a report of successful management. AB - Pyloric atresia (PA) is an uncommon anomaly that may be associated with many other congenital anomalies, the commonest of which is junctional epidermolysis bullosa (JEB). Most of the cases of PA associated with JEB (Herlitz syndrome) reported have been fatal. A case of PA associated with JEB, malrotation, and a high anorectal malformation with a rectourethral fistula, which was hitherto undescribed, was successfully managed at our institution. PMID- 10370041 TI - Inflammatory pseudotumor of the liver in Kostmann's disease. AB - A case in which inflammatory pseudotumor of the liver (IPTL) seemed to complicate severe congenital neutropenia (Kostmann's disease) is reported. IPTL is a rare entity, especially in childhood. The exact etiology of this lesion is unknown, but it is generally regarded as a benign, reactive inflammatory condition. Based on 15 reported pediatric cases in the literature, the causes, diagnosis, and treatment of IPTL are discussed. PMID- 10370042 TI - Subcapsular hemorrhage of the liver in a very-low-birth-weight neonate: survival after decompression laparotomy. AB - Subcapsular hemorrhage of the liver in a very-low-birth-weight neonate was successfully treated by decompression laparotomy. This may be the second smallest survivor after surgery in the literature. PMID- 10370043 TI - Megacystis-microcolon-intestinal hypoperistalsis syndrome associated with megaesophagus. AB - Megacystis-microcolon-intestinal hypoperistalsis syndrome is a rare congenital disorder characterized by megacystis and hypoperistalsis of the gastrointestinal tract. About 80 cases have been reported, predominantly in females. We present a female newborn with typical features of the syndrome associated with megaesophagus. PMID- 10370044 TI - Urachal cyst associated with a suprapubic sinus. AB - Urachal anomalies are frequent and may exhibit many anatomical variations. We report a case of a urachal cyst that had a sinus tract extending to the lower abdomen just above the pubic symphysis. Histologic examination of the specimens showed a squamous-epithelium-lined sinus tract and a columnar-epithelium-lined cyst, which suggested a developmental disorder. This may be an extremely rare case where the urachal cyst opened into the suprapubic sinus. PMID- 10370045 TI - Gartner's duct cyst with unilateral renal dysplasia presenting as an introital mass in a new born. AB - A persistent Gartner's duct cyst associated with ipsilateral renal agenesis or dysplasia is rare. A vaginal cyst at the introitus as the presenting complaint is very rare, and has not been previously described in a neonate. Sepsis despite the presence of renal agenesis, or non- or poorly functioning renal tissue, is an indication for ureterectomy or nephroureterectomy on the affected side. PMID- 10370046 TI - Extensive epidural teratoma in early infancy treated by multi-stage surgery. AB - We report a rare case of extensive extradural teratoma successfully treated by multi-stage laminotomy and thoracotomy. A 34-day-old, dyspneic infant had a large posterior mediastinal mass identified on a chest X-ray radiograph. Imaging studies disclosed that the mass originated from the extradural space at the level of the lower thoracic spine, extending cephalad to C4 and caudad to L4 and severely compressing the spinal cord anteriorly, causing paraplegia. The tumor expanded bilaterally through the intraspinal foramina, coalescing to form a huge mediastinal mass. The upper half of the teratoma was removed utilizing a laminotomy from T3 through T9; 2 months later the lower half was excised via a laminotomy from T11 to L3. An additional procedure was required to resect recurrent tumor through a laminotomy from T8 to T12. The reconstructed vertebral arches were well-preserved in shape, with an almost normal spinal canal. PMID- 10370047 TI - Intrapericardial teratoma diagnosed prenatally in a twin fetus. AB - Prenatal diagnosis at 32 weeks' gestation of an anteromediastinal tumor in a twin fetus allowed immediate neonatal intensive management after delivery at 34 weeks' gestation. At 48 h of age the patient underwent a median sternotomy; complete resection of the tumor was possible. Histologically, it was a mature teratoma. At age 1 year both twins are well. PMID- 10370048 TI - Magnetic resonance imaging helps in the early diagnosis of myositis ossificans in children. AB - Two cases on myositis ossificans circumscripta (MOC) in the arm are reported. Plain X-ray films and magnetic resonance imaging (MRI) were performed in both cases. In the first, an intramuscular tumor-like mass without calcifications was found on MRI with soft-tissue edema extension. In the second, MRI disclosed additional bone-marrow edema. The diagnosis of MOC was confirmed by biopsy in one case and by follow-up in the other. MOC is a benign soft-tissue lesion that is rare in children, with an acute course and usually spontaneously favorable evolution. The differential diagnosis from an infection or a malignant tumor remains difficult. The best imaging modalities are conventional radiography and MRI. The MRI patterns of MOC are typical but not pathognomonic; typical MRI findings in conjunction with clinical symptoms during the early phase of MOC permit the postponement of a biopsy or aggressive surgical procedures. Surgery is indicated for cases not showing typical MOC calcifications at a later stage. PMID- 10370049 TI - Bladder-neck repair in urinary bladder exstrophy. AB - A simple modification of an existing technique for bladder-neck reconstruction in exstrophy of the urinary bladder is reported. The technique involves tubularization of the posterior urethra up to just below the ureteric orifices. It differs from other techniques in that no part of the bladder tissue is used for buttressing the repair, but all is utilized for enhancing the bladder volume. Only 2 of 20 patients remained incontinent after bladder-neck reconstruction; the remaining 18 have achieved socially acceptable continence. PMID- 10370050 TI - Primary reconstruction of a congenital anterior urethral diverticulum. AB - A new technique for primary reconstruction of a congenital anterior urethral diverticulum is described. This operation eliminates the obstruction caused by the distal lip of the diverticulum and at the same time strengthens the unsupported urethral wall using tissue from the walls of the diverticulum. It has been used very effectively in two boys. PMID- 10370051 TI - Use of a flexible, illuminated stylet to demonstrate the proximal oesophageal pouch in surgical repair of oesophageal atresia. PMID- 10370052 TI - Comparison of hepaticoantrostomy and hepaticojejunostomy. PMID- 10370053 TI - Upper-pole pelviureteric junction obstruction: a critical review. PMID- 10370054 TI - The use of laparoscopy in the diagnosis and treatment of ventriculo-peritoneal shunt in children. PMID- 10370055 TI - The CLC chloride channel family. AB - Chloride channels perform important roles in the regulation of cellular excitability, in transepithelial transport, cell volume regulation, and acidification of intracellular organelles. This variety of functions requires a large number of different chloride channels that are encoded by genes belonging to several unrelated gene families. The CLC family of chloride channels has nine known members in mammals that show a differential tissue distribution and function both in plasma membranes and in intracellular organelles. CLC proteins have about 10-12 transmembrane domains. They probably function as dimers and may have two pores. The functional expression of channels altered by site-directed mutagenesis has led to important insights into their structure-function relationship. Their physiological relevance is obvious from three human inherited diseases (myotonia congenita, Dent's disease and Bartter's syndrome) that result from mutations in some of their members and from a knock-out mouse model. PMID- 10370057 TI - Sarcomere length during contraction of isolated guinea-pig ventricular myocytes. AB - An improved method was developed for measuring sarcomere length (SML) during twitch contractions of single cardiac ventricular myocytes, using a charge coupled photodiode array self-scanning at a rate of 1.5 ms/element. The average resting SML of 111 cells was 1.88+/-0.04 microm (mean +/-SD). When contractions were triggered by action potentials under perforated-patch conditions, the time course of SML shortening closely followed changes in cell length. A large variation was observed in contraction time course between myocytes, some cells having a phasic component with a duration at 50% shortening (full-width at half maximum; FWHM) of approximately 40 ms, while others shortened more slowly (FWHM of phasic component @100 ms). FWHM was highly correlated with relaxation half time, but with neither action potential duration nor resting SML. The kinetics of slowly contracting cells could not be converted to the rapid type by using conditioning trains or applying isoprenaline. The steady-state SML/pCa relation in ventricular myocytes was measured by applying solutions of various pCa immediately after localized punctures of the surface membrane using a focal laser beam. The Hill coefficient, nH, was @4-5 and K1/2@400-500 nM, but there was no evidence of two populations of cells with different Ca2+ sensitivities. PMID- 10370056 TI - Lack of involvement of G proteins in the activation of cardiac CFTR Cl- current by genistein. AB - The involvement of guanine nucleotide-binding proteins (G proteins) in the activation of cardiac adenosine 3',5'-cyclic monophosphate (cAMP)-dependent cystic fibrosis transmembrane conductance regulator (CFTR) Cl- current (ICl) by the tyrosine kinase inhibitor genistein (GST) was investigated in guinea-pig ventricular myocytes. Pertussis toxin (PTX) and intracellular application of 1 mM non-hydrolysable guanosine-5'-0-(2-thiodiphosphate) (GDPbetaS) and guanosine-5'-0 (3-thiotriphosphate) (GTPgammaS) were used to modify G protein activity, and the efficacy of the treatments determined by examining the activation of ICl by isoproterenol (ISO) and forskolin (FSK), and its inhibition by 1 microM acetylcholine (ACh). GDPbetaS inhibited ISO-activated ICl by 80-90%, but had little effect on ICl activated by different GST regimens (50 microM; 100 microM; 50 microM plus 0.1 microM FSK). GTPgammaS had little effect on the amplitude of ICl activated by 1 microM ISO, whereas it increased the amplitude of the current activated by 50 and 100 microM GST and rendered it insensitive to 1 microM ACh (inhibition of 2+/-2% versus (PTX-sensitive) inhibition of 94+/-3% in control myocytes). Unlike ICl activated by ISO in GTPgammaS-dialysed myocytes, ICl activated by GST deactivated on removal of the drug. GST (50 microM) reversibly increased ICl by nearly 50% in myocytes with Gs selectively activated by 1 microM ISO, and also reversibly increased the ICl that was persistently activated after withdrawal of ISO from GTPgammaS-dialysed myocytes. These results indicate that G proteins are not involved in the pathway between GST binding and CFTR opening, and suggest that enhanced adenylate cyclase activity in GTPgammaS-dialysed myocytes mediates the potentiated responses to GST. PMID- 10370058 TI - Anion selectivity of apical membrane conductance of Calu 3 human airway epithelium. AB - Anion selectivity of the cystic fibrosis conductance transmembrane conductance regulator (CFTR) and other channels and parallel pathways expressed endogenously in apical membranes of polarized Calu-3 epithelial monolayers was studied under control conditions and during cAMP stimulation. Basolateral membranes were eliminated using alpha-toxin. The cAMP-stimulated, gradient-driven currents had the sequence Br>/=Cl>/=NO3>SCN> I>/=F>formate>HCO3>acetate>propionate=butyrate=ATP= PPi=PO4=SO4=0. Selectivity of parallel cAMP-independent pathway(s) was Br>Cl=SCN=NO3>I>formate=F >HCO3>acetate>propionate. SCN, I, F or formate blocked cAMP-stimulated, but not control, Cl currents. Anions >0.53 nm in diameter were impermeant, suggesting that the apical CFTR channel has a limiting diameter of 0.53 nm. The selectivity, blocking patterns and pore size of the cAMP-stimulated conductance pathway were very similar to those in previous reports in which CFTR was heterologously expressed in non-epithelial cells. Thus, CFTR appears to be the major apical anion conductance pathway in Calu-3 cells, and its conduction properties are independent of the expression system. CFTR in Calu-3 cells also conducts physiologically relevant anions, but not ATP, PO4 or SO4. A pathway parallel (probably a tight junction) showed a different selectivity than CFTR. PMID- 10370059 TI - Vestibular semicircular canal epithelium of the rat in culture on filter support: polarity and barrier properties. AB - The inner ear of mammals contains the vestibular apparatus which is involved in the maintenance of posture and balance. The tubular structure of the apparatus is bathed by the potassium-rich endolymph and sodium-rich perilymph in the luminal and abluminal compartments, respectively. The luminal compartment is lined by a continuous epithelium with islets of receptor organs, which separates the luminal from the abluminal compartment. The present work focuses on the epithelium, without the receptor organs, and shows that it can be reconstituted in culture. The epithelium from 4-day-old Wistar rats was grown on microporous membranes. High transepithelial electrical resistances (4000-6000 Omega.cm2) were achieved after 4-8 days in culture. The epithelium was characterized by the presence of cytokeratin, ZO-1 protein, occludin, and the presence of tight junctions and kinocilia. The transepithelial resistance of the cell monolayer withstood endolymph/perilymph dual bathing when the apical pole of the cells was in contact with endolymph, but collapsed in the reverse configuration. Weak but statistically highly significant basal to apical rubidium (86Rb) transport was observed. These findings show that this epithelium maintains its in vivo polarity and could enhance the potassium composition of endolymph up to maturity. This new culture model, in which dual bathing is possible, should enable further in vitro studies of the sensory vestibular epithelia. PMID- 10370060 TI - A Na+-activated K+ current (IK,Na) is present in guinea-pig but not rat ventricular myocytes. AB - The effects of removing extracellular Ca2+ and Mg2+ on the membrane potential, membrane current and intracellular Na+ activity (aiNa) were investigated in guinea-pig and rat ventricular myocytes. Membrane potential was recorded with a patch pipette and whole-cell membrane currents using a single-electrode voltage clamp. Both guinea-pig and rat cells depolarize when the bathing Ca2+ and Mg2+ are removed and the steady-state aiNa increases rapidly from a resting value of 6.4+/- 0.6 mM to 33+/-3.8 mM in guinea-pig (n=9) and from 8.9+/-0.8 mM to 29.3+/ 3.0 mM (n=5) in rat ventricular myocytes. Guinea-pig myocytes partially repolarized when, in addition to removal of the bathing Ca2+ and Mg2+, K+ was also removed, however rat cells remained depolarized. A large diltiazem-sensitive inward current was recorded in guinea-pig and rat myocytes, voltage-clamped at 20 mV, when the bathing divalent cations were removed. When the bathing K+ was removed after Ca2+ and Mg2+ depletion, a large outward K+ current developed in guinea-pig, but not in rat myocytes. This current had a reversal potential of 80+/-0.7 mV and was not inhibited by high Mg2+ or glybenclamide indicating that it is not due to activation of non-selective cation or adenosine triphosphate (ATP)-sensitive K channels. The current was not activated when Li+ replaced the bathing Na+ and was blocked by R-56865, suggesting that it was due to the activation of KNa channels. PMID- 10370061 TI - Hypoxia inhibits the synthesis of phosphoinositides in the rabbit carotid body. AB - Hypoxic transduction in the carotid body (CB) is regulated by several systems of second messengers, but the role of the phospholipase C system has not been studied. The aim of the present study was to characterize the turnover rate of inositol phosphates (InsPs) and phosphoinositides (PIs) and their modifications by hypoxia in the rabbit CB in vitro. In CBs, in which the PIs had been labelled previously with 3H-myo-inositol, hypoxia in the presence of LiCl did not modify the accumulation of 3H-InsPs, whilst exposure to hypoxia during the loading period in the presence of LiCl reduced the accumulation of 3H-InsPs by more than 50%. Endogenous levels of inositol 1,4,5-trisphosphate were unaltered by hypoxia. Synthesis of 3H-PIs from 3H-myo-inositol was markedly inhibited by hypoxia in the CB, but not in the rat superior cervical ganglion used as control tissue. Levels of 3H-phosphatidylinositol (3H-PtdIns), 3H-phosphatidylinositol 4-monophosphate and 3H-phosphatidylinositol 4,5-bisphosphate were similarly decreased, indicating that inhibition occurs at a step prior to PtdIns synthesis. It is concluded that the phospholipase C system of second messengers does not play a significant role in the short-term regulation of hypoxic transduction cascade. It can be speculated that the decrease in PI availability produced by hypoxia might be involved in the functional changes observed in the CB on chronic hypoxic exposure. PMID- 10370062 TI - Electrophysiological properties of gap junction channels in hepatocytes isolated from connexin32-deficient and wild-type mice. AB - Hepatocytes were isolated from wild-type and connexin32-deficient (Cx32 deficient) mice. Pairs of cells were chosen to study the electrical properties of gap junction channels using the dual voltage-clamp method. The total gap junction currents revealed that Cx32-deficient hepatocytes express one type of connexin (Cx26) and wild-type hepatocytes express two types of connexins (Cx26 and Cx32). The unitary gap junction currents suggest that Cx32-deficient cells have homotypic channels (Cx26-Cx26) while wild-type cells form homotypic (Cx26-Cx26, Cx32-Cx32) and heterotypic channels (Cx26-Cx32). Homotypic channels exhibited a main conductance and a residual conductance, both virtually insensitive to gap junction voltage (Vj) (Cx32-Cx32: gammaj,main=31 pS, gammaj,residual=9 pS; Cx26 Cx26: gammaj,main=102 pS, gammaj,residual=17 pS). Residual states were regularly seen in Cx32-Cx32 channels, but rarely in Cx26-Cx26 channels. Heterotypic channels showed a main conductance and a residual conductance. The former was sensitive to Vj (average gammaj,main=52 pS). The electrophysiological data suggest that Cx32 hemichannels are more abundant than Cx26 hemichannels in prenatal (ratio 4:1) and adult wild-type hepatocytes (ratio 23:1) and that the total number of gap junction channels is larger in prenatal cells than in adult cells. The diversity of the relationship gj, ss/gj,inst=f(Vj) (gj,ss: gap junction conductance at steady state; gj,inst: instantaneous gap junction conductance; Vj: transjunctional voltage) seen in wild-type cells suggests that the ratio Cx26/Cx32 hemichannels is variable among hepatocytes. A comparison of total and unitary conductances implies that Cx26 hemichannels are down-regulated in Cx32-deficient cells and that docking between Cx26 and Cx32 hemichannels occurs randomly. While the gap junction currents are compatible with homotypic and heterotypic channels, the presence of heteromeric channels cannot be excluded. PMID- 10370063 TI - Increased mRNAs for procollagens and key regulating enzymes in rat skeletal muscle following downhill running. AB - The purpose of the study was to investigate pre-translational regulation of collagen expression after a single bout of exercise. We analysed steady-state messenger ribonucleic acid (mRNA) levels for collagen types I, III and IV, alpha- and beta-subunits of prolyl 4-hydroxylase and lysyl oxidase (enzymes modifying procollagen chains), and enzyme activity of prolyl 4-hydroxylase from rat soleus muscle (MS) and the red parts of quadriceps femoris muscle (MQF) after 12 h and after 1, 2, 4, 7 and 14 days of downhill (-13.5 degrees ) treadmill running at a speed of 17 m.min-1 for 130 min. Histological and biochemical assays revealed exercise-induced muscle damage in MQF but not MS. Steady-state mRNA levels for the alpha- and beta-subunits of prolyl 4-hydroxylase in MQF, lysyl oxidase in MS and MQF were increased 12 h after running, whereas prolyl 4-hydroxylase activity did not increase until 2 days after exercise. The mRNA levels for the fibrillar collagens (I and III) and basement membrane type IV collagen significantly increased 1 day and 12 h after exertion, respectively. Peak mRNA levels were observed 2-4 days after running, the increases being more pronounced in MQF than in MS. No significant changes were observed in types I or III collagen at the protein level. Strenuous downhill running thus causes an increase in gene expression for collagen types I and III and their post-translational modifying enzymes in skeletal muscle in a co-ordinated manner. These changes, together with the increased gene expression of type IV collagen, may represent the regenerative response of muscle extracellular matrix to exercise-induced injury and an adaptive response to running exertion. PMID- 10370064 TI - H+ ion modulation of C-type inactivation of Shaker K+ channels. AB - The participation of an extracellular loop in C-type inactivation of voltage gated K+ channels was investigated. A wild-type phenylalanine (at position 425) between the fifth putative transmembrane segment (S5) and the pore region of the Shaker K+ channel was mutated to a histidine and the functional consequences of protonating the imidizole group of the histidine were examined. C-type inactivation of both wild-type and mutant channels was sensitive to external pH over the range of 5.2-8. The pH dependence of wild-type channels was characterized by an apparent pK value of 5. 0. The inactivation kinetics of F425H mutant channels had a pH dependence with a pK of 5.8 - in addition to the pH dependence of the wild-type channels. Moreover, at pH 7 and 8 the voltage dependence of C-type inactivation kinetics was manifest only in the F425H mutant channels. C-type inactivation in wild-type channels involves a chemical group with a low pK. Taken together, these results suggest that residues located in the extracellular S5-pore loop of the Shaker K+ channel participate in C-type inactivation. PMID- 10370065 TI - Properties of T-type calcium current in enkephalinergic neurones in guinea-pig hypothalamic slices. AB - The guinea-pig hypothalamic magnocellular dorsal nucleus (mdn) exclusively contains enkephalinergic neurones providing inputs to the septum. This nucleus is believed to play a role in hippocampo-septo-hypothalamic relationships. mdn neurones display prominent low-threshold Ca2+ spikes, which differ in their propensity to trigger either a burst of Na+ spikes or a single spike. In the present study, whole-cell voltage-clamp experiments were carried out on thick slices at 34 degrees C to characterize the pharmacological and physical properties of the transient Ca2+ current (IT) underlying the low-threshold spikes. Recorded cells were dye-labelled and identified as belonging to the mdn. In bursting and non-bursting neurones, IT was reduced by amiloride (1 microM) and octanol (1 mM), and during replacement of Ca2+ by Ba2+. The Ca2+ channel blocker mibefradil (10 microM) had only a slight blocking action. Nifedipine (100 microM) and flunarizine (1 microM) had no effect. IT activated between -80 mV and -50 mV and the mean peak current was 1050 pA. Steady-state activation and inactivation curves were fitted by a Boltzmann equation. The half-activation voltage was -70 mV, slope factor=3.6, and half-inactivation voltage was about -80 mV, slope factor=4.5. Time-to-peak and time constant of inactivation were voltage dependent. Recovery from activation occurred within 500 ms. When compared with results on other IT, the present data show that the current possesses distinct pharmacological and physical properties. Nevertheless, all investigated cells displayed a homogenous profile of IT, suggesting that the differences in spike pattern between mdn neurones are not due to different populations of Ca2+ channels. PMID- 10370066 TI - AT1 receptor up-regulation in intestine in chronic renal failure is segment specific. AB - The role of the AT1 receptor in stimulating colonic K+ secretion was investigated in rats with chronic renal failure (CRF) induced by 5/6 nephrectomy. Compared to control rats, CRF rats up-regulate mucosal AT1 receptors approximately two-fold in the proximal (PC) and distal (DC) colonic segments. In contrast, there was no alteration in AT1 receptor protein mass in jejunal or ileal mucosa. Using 86Rb+ as a tracer for transmural K+ fluxes, a significant stimulation of the basal K+ secretory flux across both PC and DC was observed in vitro and this alteration in K+ transport was temporally correlated with the increase in angiotensin II (ANG II) receptors. In both PC and DC the significant increases in the receptor protein were evident 48 h after partial nephrectomy and they were sustained through 6 weeks. These studies support the hypothesis that CRF-induced secretion of K+ is mediated by an up-regulation of AT1 receptors exclusively in the large intestinal segments. PMID- 10370067 TI - Complete reversal of run-down in rabbit cardiac Ca2+ channels by patch-cramming in Xenopus oocytes; partial reversal by protein kinase A. AB - The rabbit cardiac Ca2+ channel (alpha1C) expressed in Xenopus oocytes exhibited a complete run-down of ionic currents when cell-attached patches were excised. The alpha1C channel was expressed alone or was coexpressed with the accessory beta2a or beta1b subunit. The catalytic subunit of protein kinase A (PKAc) and MgATP were capable of delaying the run-down of single-channel currents. In 33% of the alpha1C patches, and 26% of the alpha1C+beta2a patches, inclusion of PKAc in the bath solution delayed the run-down for a maximum of 20 min. In experiments where PKAc in the bath was not sufficient to delay the run-down of channel activity, insertion of the patch back into the oocyte (patch-cramming) could restore channel activity. Gating currents were also measured in the alpha1C+beta1b channel and were not subject to any run-down, even after the complete run-down of ionic currents. The results presented here reveal that PKAc is capable of delaying the run-down of currents in a subset of patches. The patch cramming results suggest that a cytoplasmic factor, in addition to phosphorylation of the channel (by PKAc), may be involved in the maintenance of channel activity. PMID- 10370068 TI - Localization of sodium and potassium currents at sites of nerve-muscle contact in embryonic Xenopus muscle cells in culture. AB - Macro-patch current recordings were used to determine the spatial distribution of INa, inactivating (IIK) and non-inactivating (IK) outward potassium currents on nerve-contacted Xenopus muscle cells grown in culture. Sites of nerve contact were identified under 400x magnification and macroscopic currents recorded from cell-attached membrane patches ranging in diameter from 2 microm to 6 microm. Membrane patch (n=133) recordings revealed no detectable current, or INa and/or IIK, or just IK. More than 85% of the membrane patches from which INa (n=35) could be recorded occurred within 40 microm of the position of nerve contact. In addition, more than 90% of the membrane patches which contained INa also contained IIK. The density of IIK in these membrane patches that also contained INa was significantly higher than the density of IIK or IK in membrane patches without INa. Sites of ACh receptor localization at nerve-muscle contacts, identified by labelling with rhodamine-alpha-bungarotoxin, were also found to contain both INa (4/5 cells) and IIK (5/5 cells). These results suggest that in Xenopus muscle cells in culture the channels mediating IIK along with sodium channels are clustered at sites of nerve contact and synaptic specializations. PMID- 10370070 TI - Cardioventilatory coupling in the anaesthetised rabbit, rat and guinea-pig. AB - Cardioventilatory coupling is a temporal coherence of respiratory and cardiac rhythms, seen in humans at rest, and during sleep and anaesthesia. In this study we compared the cardioventilatory coupling of anaesthetised rabbits, rats and guinea-pigs. Breathing two successive anaesthetic concentrations (1 or 2% isoflurane) we compared the effect of anaesthetic depth and species on (1) heart rate, (2) heart rate variability, (3) ventilatory rate (f), (4) ventilatory variability, (5) ratio HR/f, (6) degree of coupling (Shannon entropy of the distribution of intervals between inspiration and the preceding electrocardiographic R wave - the RI interval) and (7) coupling pattern, classified into four sub-patterns (I-IV) based upon inspection of the RI interval time series. Rabbits exhibited significantly less ventilatory variability and coupling than rats or guinea-pigs. The sub-pattern of coupling also differed between the three species. Rabbits showed coupling only when HR and f were close to integer ratios whereas other species coupled at non-integer ratios. Ventilatory variability in the rat and guinea-pig differed according to the pattern of coupling observed. Of the three species studied, the rat and guinea pig demonstrated coupling most similar to that of anaesthetised human subjects. Anaesthetic concentration did not influence the pattern or degree of coupling. PMID- 10370069 TI - 5-hydroxytryptamine blocks the fetal more potently than the adult mouse muscle acetylcholine receptor. AB - The action of 5-hydroxytryptamine (5-HT) on fetal (gamma-AChR) and adult (epsilon AChR) muscle acetylcholine receptors was studied in transfected BOSC 23 cells expressing mouse AChRs and in acutely dissociated mouse muscle fibres. In transfected cells, coapplication of 5-HT (0.01-10 mM) with ACh reversibly reduced the amplitude and accelerated the decay of the ACh-activated whole-cell currents, in a dose- and voltage-dependent manner. 5-HT blocked faster and with higher potency the gamma-AChR than the epsilon-AChR. Cell-attached recordings from transfected BOSC 23 cells and embryonic or neonatal muscle fibres were made. When 5-HT (5 microM) was included in the patch pipette, ACh-evoked unitary events acquired a bursting behaviour, which was more pronounced for gamma- than epsilon AChR, though it was enhanced by membrane hyperpolarization for both AChR species. There was no effect on the single-channel conductance. It is concluded that 5-HT behaves as an open-channel blocker of muscle AChRs, with higher efficacy on gamma than on epsilon-AChR. PMID- 10370071 TI - Novel properties of the depolarization-induced endogenous sodium conductance in the Xenopus laevis oocyte. AB - It has been shown by means of the two-microelectrode voltage-clamp technique that in membranes of Xenopus laevis oocytes a Na+-selective permeability can be activated by long-lasting or repetitive depolarization (R.T. Kado and C. Baud, Journal of Physiology, Paris, 77:1113-1117, 1981). In this study the permeability in inside-out giant membrane patches with diameters of 20-30 microm was analysed. Once induced, the Na+ permeability has a voltage-dependent open probability that increases with positive potentials and half-maximally activates at about 0 mV. Sudden changes of membrane potential elicit transient currents with strongly voltage-dependent time constants of from less than 1 ms at -150 mV to several hundreds of milliseconds at positive potentials. In contrast to the on-cell configuration, the permeability ceases completely within a few minutes in the cell-free inside-out configuration. This rundown can be prevented by including MgATP, but not Mg2+ or ATP alone, in the intracellular solution. Intracellular Mg2+ ions, in addition to being a co-factor for ATP in the activation process, decrease the permeability in a dose-dependent manner. Steady-state fluctuation analysis gave no evidence that an increased noise level is caused by open-close kinetics of an ion channel, suggesting that the single-channel conductance is below 1 pS if a channel-like structure is the origin of the endogenous Na+ permeability. PMID- 10370072 TI - Use of electrochemical impedance measurements to monitor beta-adrenergic stimulation of bovine aortic endothelial cells. AB - Due to the high permeability of endothelial cell layers derived from macrovascular vessels, precise determination of their barrier function towards ion movement requires refined experimental techniques. We thus cultured bovine aortic endothelial cells (BAEC) directly on thin gold-film electrodes and measured the electrochemical impedance to study their passive electrical properties in general and during beta-adrenergic stimulation. Impedance spectra (10-2.10(6) Hz) of confluent cell monolayers revealed that the electrical characteristics of the cells can be modelled by a simple resistor-capacitor parallel network. Under control conditions the overall resistance of confluent BAEC monolayers was 3.6+/-0.6 Omega.cm2 (n=30) and the capacitance was 0. 6+/-0.1 microF/cm2. Both quantities are discussed with respect to morphological characteristics of these cells. Stimulation of BAECs with the synthetic beta adrenoceptor agonist isoproterenol leads to a concentration-dependent, highly specific increase of the cell layer resistance characterized by a concentration for half-maximal response (EC50) of 0.3+/-0.1 microM. The cell layer capacitance, however, remained unaffected. Using impedance measurements at a single frequency, we analysed the response of BAECs to treatment with isoproterenol in comparison with several chemically unrelated compounds known to stimulate the adenosine 3',5'-cyclic monophosphate (cAMP)-dependent signal transduction cascade. These studies confirmed that the enhancement of the cell layer resistance after beta adrenergic stimulation is mediated by an increase in intracellular cAMP. PMID- 10370074 TI - Comparison of Na+-Ca2+ exchange current elicited from isolated rabbit ventricular myocytes by voltage ramp and step protocols. AB - In this study, the effects of three different voltage protocols on the Na+-Ca2+ exchange current (INa-Ca) of rabbit right ventricular myocytes were studied. Whole-cell patch-clamp recordings were made using a Cs+-based internal dialysis solution and external solutions designed to block major interfering currents. INa Ca was measured at 35-37 degrees C as (5 mM) Ni-sensitive current elicited by: a 2 s descending ramp (DR: +80 to -120 mV); a 2 s ascending ramp (AR: -120 to +80 mV) and 500 ms voltage steps (VS) between -120 and +80 mV. DR and AR were applied from -40 mV and elicited INa-Ca with reversal potentials (Erev) of -17.6+/-2.5 mV (mean+/-SEM; n=16) and -46.2+/-4. 1 mV (n=10; P=0.0001) respectively. This difference was maintained when the holding potential was -80 mV (-44.0+/-2.1 mV, n=24 and -86. 3+/-4.8 mV, n=10; P=0.0001), when the internal Ca chelator (EGTA) was replaced with BAPTA (-19.5+/-1.8 mV and -46.3+/-1.6 mV, n=6; P=0. 0003) and when DR and AR were applied alternately to the same cell. Experiments using modified ramp waveforms suggested a possible mechanism for these differences. Increases in subsarcolemmal Ca caused by Ca entry (coupled to Na extrusion) during the initial positive potential phase of the DR might have induced INa-Ca reversal at less negative potentials than observed with AR, during the initial phase of which subsarcolemmal Ca would not have accumulated. These data suggest that INa-Ca during voltage-clamp experiments can be significantly influenced by the type of voltage protocol chosen, as the protocol appears to induce subsarcolemmal changes in Ca and Na concentration that are independent of Ca buffering in the bulk cytosol and can occur on a pulse-to-pulse basis. PMID- 10370073 TI - Evidence for an electrogenic, negatively protein-kinase-A-modulated, Na+ dependent HCO3- transporter in human lymphocytes. AB - We studied the effects of external HCO3- on pHi regulation in human lymphocytes after an acid load. Cells were acidified by preincubation with NH4Cl and pHi recovery was measured with the fluorescent dye BCECF. Cells recovering in HCO3- containing medium reached a higher final pHi, the H+ efflux rate was increased and shifted to alkaline pHi compared to that of cells recovering in HCO3--free solution. The resting pHi was higher in a HCO3--containing solution. Experiments carried out in the presence of amiloride, DIDS and in the absence of external Na+ suggest the existence of two major mechanisms acting in the pHi recovery of lymphocytes after an acid load: an amiloride-sensitive Na+/H+ exchanger and a DIDS-sensitive Na+-dependent HCO3- transporter. The last mechanism could be a Na+/HCO3- cotransporter based on membrane potential changes determined with the potential-sensitive fluorescent probe bis-oxonol. Preincubation of cells with forskolin and H-89 showed protein-kinase-A-dependent downregulation of the amiloride-insensitive recovery of pHi in human lymphocytes. In summary, this paper provides functional evidence for the existence of a Na+/HCO3--dependent mechanism involved in pHi recovery in human lymphocytes following an acid load, that is electrogenic and downregulated by PKA. PMID- 10370075 TI - Effects of beta-escin and saponin on the transverse-tubular system and sarcoplasmic reticulum membranes of rat and toad skeletal muscle. AB - Mechanically skinned skeletal muscle fibres from rat and toad were exposed to the permeabilizing agents beta-escin and saponin. The effects of these agents on the sealed transverse tubular system (t-system) and sarcoplasmic reticulum (SR) were examined by looking at changes in the magnitude of the force responses to t system depolarization, the time course of the fluorescence of fura-2 trapped in the sealed t-system, and changes in the magnitude of caffeine-induced contractures following SR loading with Ca2+ under defined conditions. In the presence of 2 microg ml-1 beta-escin and saponin, the response to t-system depolarization was not completely abolished, decreasing to a plateau, and a large proportion of fura-2 remained in the sealed t-system. At 10 microg ml-1, both agents abolished the ability of both rat and toad preparations to respond to t system depolarization after 3 min of exposure, but a significant amount of fura-2 remained in sealed t-tubules even after exposure to 100 microg ml-1 beta-escin and saponin for 10 min. beta-Escin took longer than saponin to reduce the t system depolarizations and fura-2 content of the sealed t-system to a similar level. The ability of the SR to load Ca2+ was reduced to a lower level after treatment with beta-escin than saponin. This direct effect on the SR occurred at much lower concentrations for rat (2 microg ml-1 beta-escin and 10 microg ml-1 saponin) than toad (10 microg ml-1 beta-escin and 150 microg ml-1 saponin). The reverse order in sensitivities to beta-escin and saponin of t-system and SR membranes indicates that the mechanisms of action of beta-escin and saponin are different in the two types of membrane. In conclusion, this study shows that: (1) beta-escin has a milder action on the surface membrane than saponin; (2) beta escin is a more potent modifier of SR function; (3) simple permeabilization of membranes is not sufficient to explain the effects of beta-escin and saponin on muscle membranes; and (4) the t-system network within muscle fibres is not a homogeneous compartment. PMID- 10370076 TI - The effects of inhibition of the sarcolemmal Ca-ATPase on systolic calcium fluxes and intracellular calcium concentration in rat ventricular myocytes. AB - The effects of carboxyeosin, an inhibitor of the sarcolemmal Ca-ATPase, were studied on intracellular Ca and membrane currents in isolated rat ventricular myocytes. In the absence of carboxyeosin, 150-ms-duration depolarizing pulses from -40 to 0 mV resulted in an L-type Ca current on depolarization and a Na-Ca exchange "tail" current on repolarization. The calculated entry of Ca on the L type current was 1.3 times greater than the efflux via the Na-Ca exchange. The addition of carboxyeosin (20 microM) resulted in either an increase of the Na-Ca exchange current or a decrease of the L-type Ca current such that the Ca entry and efflux were exactly equal. These results suggest that, under control conditions, a carboxyeosin-sensitive Ca-ATPase contributes about 24% of the total Ca efflux from the cell and, therefore, that the sarcolemmal Ca-ATPase has a significant role in regulation of sarcolemmal Ca fluxes. PMID- 10370077 TI - Studies on the topology of the renal type II NaPi-cotransporter. AB - The rat type II sodium/phosphate cotransporter (NaPi-2) is a 85- to 90-kDa glycosylated protein located at the proximal tubular brush border membrane. Hydropathy predictions suggest eight transmembrane domains (sTM) with a large glycosylated loop between sTM 3 and sTM 4. We have studied the membrane topology of NaPi-2 expressed in oocytes. A 33-amino-acid fragment containing the FLAG epitope was inserted into seven loops connecting the sTMs and into the NH2- and COOH-ends of the protein. FLAG-antibody binding suggested that the loops connecting sTM 1 and sTM 2 as well as sTM 3 and sTM 4 are located extracellularly. Based on the lack of FLAG-antibody binding we suggest intracellular locations for the NH2- and COOH-termini and the region connecting sTM 4 and sTM 5. Immunoprecipitation studies of in vitro translated protein also suggest that the NH2-terminus is sited extracellularly. In immunohistochemical studies with NaPi-2-transfected MDCK cells, an interaction with NH2- and COOH- terminal antipeptide antibodies could only be obtained after membrane permeabilization. The presented data are an experimental documentation of the intracellular location of the NH2- and COOH-termini, and of the extracellular location of extracellular loops 1 and 2. PMID- 10370078 TI - The isolated working mouse heart: methodological considerations. AB - Our aim was to develop a working isolated murine heart model, as the extensive use of genetically engineered mice in cardiovascular research requires development of new miniaturized technology. Left ventricular (LV) function was assessed in the isolated working mouse heart perfused with recirculated oxygenated Krebs-Henseleit bicarbonate buffer (37 degrees C pH 7.4) containing 11.1 mM glucose and 0.4 mM palmitate bound to 3% albumin. The hearts worked against an afterload reservoir at a height equivalent to 50 mmHg, and heart rate was controlled by electrical pacing of the right atrium. LV pressure was measured with a micromanometer connected to a small steel cannula inserted through the apex of the heart. The experimental protocol consisted of two interventions. First, following instrumentation and stabilization, the preload reservoir was raised from a pressure equivalent of 7 to 22.5 mmHg, while pacing at 390 beats.min-1. Thereafter the height of the preload reservoir was set to 10 mmHg, and the pacing rate was varied from 260 to 600 beats.min-1. Aortic and coronary flows were measured by timed collections of effluent from the afterload line and that dripping from the heart, respectively [aortic+coronary flow=cardiac output (CO)]. Elevation of LV end-diastolic pressure (LVEDP) from approximately 5 to 10 mmHg resulted in a twofold increase in average cardiac power [product of LV developed pressure (LVDevP) and CO], whereas myocardial contractility (first derivative of LV pressure, dP/dt) and LVDevP (LV systolic pressure-LVEDP) increased only minimally (5-10%). Measured LVEDP was lower than the equivalent height of the preload reservoir by an amount that was related to the heart rate. Cardiac power, LVDevP and dP/dt were stable at heart rates up to 400 beats.min-1, but declined markedly with higher rates, consistent with the decrease in LVEDP. Thus, cardiac power was reduced to 50% of its maximum value when stimulated at approximately 500 beats.min-1, and at even higher rates there was little ejection. By systematic manipulation of the height of the preload reservoir and heart rate, we conclude that LV afterload and preload can be assessed only by high-fidelity measurement of intraventricular pressures. The heights of the afterload column and the preload reservoir are unreliable and potentially misleading indicators of LV afterload and preload. PMID- 10370079 TI - A rapid wavelength changer based on liquid crystal shutters for use in ratiometric microspectrophotometry. AB - Many of the fluorescent indicator molecules most useful for measuring intracellular concentrations of ions of biological importance, such as Ca2+ or H+, require illumination first at one wavelength, at which the fluorescence depends strongly on the concentration of the ion, and then at another wavelength (e.g. the isosbestic point), so that a ratio can be obtained. Existing wavelength changers are mechanical and involve moving filters, mirrors or gratings. These systems have the disadvantage that they introduce mechanical shocks that can interfere with simultaneous electrophysiological recording. In addition, they require special electrical driving systems and are relatively expensive, especially if they are capable of switching rapidly. We describe a new wavelength changer based on liquid crystal shutters which has the following advantages: (1) it has no mechanical moving parts; (2) it can switch rapidly (@1 ms) and in any desired pattern (off - on1 - off - on2 - off, etc.); (3) it is driven by a low power 15-V pulse; and (4) it is substantially cheaper than existing wavelength changers. Its limitations are that it does not pass wavelengths shorter than about 400 nm and transmission in the range 430-700 nm is only 20-40%. PMID- 10370080 TI - Expression of sulphonylurea receptor protein in mouse kidney. AB - The sulphonylurea receptor (SUR) is the site of action for sulphonylurea derivatives such as glibenclamide, which are widely used as oral hypoglycaemic agents. Sulphonylureas have also been shown to affect urine flow and salt excretion by the kidney; therefore, the use of these drugs may have important implications for the pharmacological manipulation of renal salt handling. The purpose of the present investigation was to increase our understanding of the possible role of SUR in the regulation of renal function by determining the distribution of SUR isoforms within mouse kidney. Immunostaining with anti-SUR antisera revealed specific staining of SUR2B in distal nephron segments of mouse kidney. A diffuse, low level staining was observed in proximal tubules in the inner cortical region. No evidence was found for the presence of SUR2B in intra renal blood vessels. Reverse-transcription polymerase chain reaction and Western blotting experiments indicated that SUR2B is the only known isoform expressed. These data demonstrate that SUR2B in mouse kidney is expressed in tubule regions that are critical in determining renal salt excretion. PMID- 10370081 TI - Ionic currents expressed in a cell line derived from the organ of Corti of the Immortomouse. AB - We have investigated the maturation of adult hair cell electrophysiology in a population of precursor cells in a conditionally immortal cell line. The cell line, UB/OC-2, from the embryonic organ of Corti of the H-2Kb-tsA58 transgenic mouse, permits cells to grow proliferatively at 33 degrees C and to differentiate at 39 degrees C. Whole-cell patch-clamp recordings showed that proliferating cells had a different electrophysiology to differentiating cells. Differentiating cells had a conditionally expressed slowly activating inward current activated by hyperpolarization. The current was not blocked by extracellular application of 0.5 mM Ba2+, but was blocked reversibly by 2 mM Cs+. The current was found to be carried by both K+ and Na+ ions (PK/PNa=2.2) and activated by 10 microM forskolin. These properties identify the slowly activating current as Ih. A proportion of proliferating and differentiating cells exhibited a voltage-gated Na+ current, INa. INa was abolished in Na-free external medium and was inhibited reversibly by tetrodotoxin (TTX) with Ki=64 nM. Together these results suggest that proliferating and differentiating hair cell precursors in the immortal cochlear cell line UB/OC-2 express currents which are also found in developing hair cells. PMID- 10370082 TI - The cellular mechanisms of Cl- secretion induced by C-type natriuretic peptide (CNP). Experiments on isolated in vitro perfused rectal gland tubules of Squalus acanthias. AB - We have examined the mechanism whereby C-type natriuretic peptide (CNP), an agonist acting through the second messenger cGMP, enhances NaCl secretion in the rectal gland of Squalus acanthias. Single rectal gland tubules (RGT) were dissected manually, perfused in vitro and equivalent short-circuit current [Isc=transepithelial voltage/transepithelial resistance (Rte)] as well as basolateral membrane voltage (Vbl) were measured. CNP was added to luminal and basolateral perfusates at concentrations between 1 and 1000 nmol/l and its effects on the above parameters were compared to those of a "stimulation cocktail" (Stim, containing dibutyryl cAMP, adenosine and forskolin) that maximally enhances cytosolic cAMP, and other agonists and hormones such as guanylin, vasoactive intestinal peptide (VIP), and adenosine. CNP had no effect from the luminal side (n=6). Its effects from the basolateral side consisted of a substantial increase in Isc (-31.6+/-7.7 to -316+/-82.2 microA/cm2, n=15). CNP significantly depolarized the luminal membrane from -87. 4+/-1.0 to -82.3+/-2.6 mV (n=12). Vbl was not changed (n=12) but the fractional conductance for K+ was increased (n=3). These effects were qualitatively and even quantitatively comparable to those of other agonists acting via cytosolic cAMP, but were less marked than those caused by Stim (n=64). The effects of VIP and CNP on Isc were not additive (n=5). The cytosolic Ca2+ concentration ([Ca2+]i) was monitored using the fura-2 fluorescence ratio (FFR 340/380 nm) and it was found that CNP, like agonists acting via cAMP, enhances FFR significantly from 1.02+/-0.05 to 1.32+/-0.05 (n=8) with a time constant in the 1-2 min in range. Our data suggest that CNP, acting via the second messenger cGMP, induces a marked increase in Isc in the rectal gland. The concomitant fall in Rte corresponds to increases in the luminal membrane Cl- conductance and in the basolateral membrane K+ conductance. The latter effect is probably due to an increase in [Ca2+]i. PMID- 10370083 TI - Blockade of HERG channels expressed in Xenopus oocytes by external H+. AB - We have investigated the effect of external H+ concentration ([H+]o) on the human ether-a-go-go-related gene (HERG) current (IHERG), the molecular equivalent of the cardiac delayed rectifier potassium current (IKr), expressed in Xenopus oocytes, using the two-microelectrode voltage-clamp technique. When [H+]o was increased, the amplitude of the IHERG elicited by depolarization decreased, and the rate of current decay on repolarization was accelerated. The activation curve shifted to a more positive potential at lower external pH (pHo) values (the potential required for half-maximum activation, V1/2, was: -41.8 mV, -38.0 mV, 33.7 mV, -26.7 mV in pHo 8.0, 7.0, 6.6, 6.2, respectively). The maximum conductance (gmax) was also affected by [H+]o: a reduction of 7.9%, 14.6%, and 22.8% was effected by decreasing pHo from 8.0 to 7.0, 6.6, and 6.2, respectively. We then tested whether this pH effect was affected by the external Ca2+ concentration, which is also known to block HERG channels. When the extracellular Ca2+ concentration was increased from 0.5 mM to 5 mM, the shift in V1/2 caused by increasing [H+]o was attenuated, suggesting that these two ions compete for the same binding site. On the other hand, the decrease in gmax caused by increasing [H+]o was not significantly affected by changing external Ca2+ levels. The results indicate that HERG channels are inhibited by [H+]o by two different mechanisms: voltage-dependent blockade (shift of V1/2) and the decrease in gmax. With respect to the voltage-dependent blockade, the interaction between H+ and Ca2+ is competitive, whereas for the decreasing gmax, their interaction is non competitive. PMID- 10370084 TI - Characterisation of a mutant of barnacle troponin C lacking Ca2+-binding sites at positions II and IV. AB - This study investigates a mutant barnacle troponin C (TnC) protein (BTnC2-4-) in which the Ca2+-binding sites (sites II and IV) have been rendered non-functional. Eliminating Ca2+ binding at both Ca2+-binding sites of barnacle TnC did not prevent the incorporation of BTnC2-4- into TnC-depleted myofibrillar bundles, although, as expected, the mutant was not able to effect muscle regulation. We conclude that the Mg2+ involved in stabilising the interaction between TnC and TnI in the barnacle must bind at a separate location to the Ca2+-binding sites. Competition experiments between BTnC2-4- and wild-type barnacle TnC (BTnCWT) or the native isoform BTnC2 indicate that BTnC2-4- has an approximately fourfold higher affinity for barnacle TnI than BTnCWT but a lower affinity for TnI compared to BTnC2. These results indicate that disabling sites II and IV changes the affinity of BTnC2-4- for TnC-denuded barnacle myofibrils, altering the stability of the bond formed between TnC and the thin filament. PMID- 10370085 TI - Ionic currents and current-clamp depolarisations of type I and type II hair cells from the developing rat utricle. AB - Ionic currents and the voltage response to injected currents were studied in an acutely dissected preparation of the rat utricle between birth and postnatal day 12 (PN12). Based upon morphological criteria, the sensory cells examined were divided into two classes, "type I" and "type 2 category," the latter of which may include some immature type I cells. The former group comprises a clearly defined electrophysiological population, with one large outwardly rectifying potassium conductance that is sensitive to 4-aminopyridine (4-AP), insensitive to tetraethylammonium (TEA) and displays voltage-dependent activation kinetics. In the absence of enzymatic dissociation procedures, and with the epithelium left largely intact, the mean half activation of this conductance was -30.3 mV at PN3, and -37.5 mV at PN12. At both stages it was almost entirely turned off at -74 mV. Omission of ATP from the intracellular solution appeared to prevent rundown of this conductance. Type II category hair cells formed a more heterogeneous population, exhibiting a distinct TEA-sensitive delayed rectifier potassium conductance; the rapidly activating and inactivating IA; an inward rectifier; and inward sodium currents at around PN3. Both cell types depolarised strongly in response to injected currents, with time courses reflecting the activation kinetics of their major outward conductances. PMID- 10370086 TI - The influence of the paraventricular nucleus on baroreceptor dependent caudal ventrolateral medullary neurones of the rat. AB - The question of whether neurones in the paraventricular nucleus (PVN) of the hypothalamus have an influence on barosensitive vasomotor neurones of the caudal ventrolateral medulla (CVL) was addressed in this study using anaesthetised rats. Extracellular microelectrode recordings were made from 27 vasomotor neurones in the CVL, identified by their cardiac cycle-related probability of discharge, by the increase in activity in response to an increase in arterial blood pressure produced by i.v. phenylephrine and by the decrease in activity in response to a decrease in blood pressure produced by i.v. nitroprusside. Activation of neurones at different sites in the PVN with a microinjection of d,l-homocysteic acid (DLH) activated 12 CVL neurones and inhibited 6 CVL neurones. In four CVL units single shock electrical stimulation at a PVN pressor site, first identified with DLH, elicited a synaptic action potential with a latency of 29+/-0.3 ms. It is concluded that PVN neurones can influence the CVL vasomotor neurones directly. This could be one means by which PVN-depressor and sympatho-inhibitory effects are produced. PMID- 10370087 TI - Bradykinin-stimulated Cl- secretion in T84 cells. Role of Ca2+-activated hSK4 like K+ channels. AB - Bradykinin (BK)-stimulated colonic Cl- secretion was studied in T84 colonic adenocarcinoma cells by measuring BK (50 nM)-evoked changes in cytosolic free [Ca2+] ([Ca2+]i), membrane conductance and transepithelial ion transport. In T84 cells grown on impermeable supports, BK stimulated a transient increase in [Ca2+]i as assessed by fura-2 ratio imaging. In cell-attached, patch-clamp recordings, BK transiently activated low-conductance K channels. These channels were activated/inactivated reversibly in inside-out patches by switching [Ca2+]i in the bath between 30 nM and 100 nM. Excised channels recorded with 160 mM [K+] in bath and pipette exhibited an inwardly rectifying current/voltage-relation, conductances of 10+/-1 pS and 34+/-4 pS (n=10) at positive and negative voltages, respectively, and a 15-fold lower permeability for Na+ than for K+. The mean open probability of these channels did not depend on voltage but increased with increasing [Ca2+]i with an apparent concentration for a half-maximal response (EC50) of 110 nM, resembling that of hSK4 K+ channels. Application of the reverse transcriptase-polymerase chain reaction technique showed hSK4 messenger ribonucleic acid (mRNA) to be expressed in T84 cells. Macroscopic currents in T84 cells showed a similar dependence on [Ca2+]i. Whole cell conductance (in nS/10pF) increased from 0.5+/-0. 1 (n=6) at 10 nM [Ca2+]i in the pipette solution to 1.5+/ 0.2 (n=7) at 100 nM, and to 2.0+/-0.5 (n=7) at 1 microM due to activation of a K+ conductance. In Ussing-chambered T84 monolayers grown on filters, BK did not evoke a short-circuit current (Isc). When, however, the monolayers were pre stimulated by forskolin (1 microM), BK further enhanced Cl-secretion (DeltaIsc=21+/-5 microA/cm2, n=10) transiently and biphasically. In conclusion, BK enhances cyclic adenosine monophosphate-stimulated Cl- secretion in T84 cells, probably via basolateral, Ca2+-liganded activation of low-conductance hSK4-type K+ channels. PMID- 10370088 TI - Effects of phorbol esters on excitation-contraction coupling and protein kinase C activity of frog twitch muscle fibers. AB - By recording the calcium transients evoked by voltage-clamp depolarizing pulse with arsenazo III as a calcium indicator, it has been shown that 1 micromol/l phorbol 12,13-dibutyrate (PDBu), a protein kinase C (PKC) agonist, causes a transient potentiation and then a depression of the calcium transients of twitch muscle fibers in frogs. PDBu also produces an initial translocation and activation of PKC, which is followed by a down-regulation. To find out whether the effect of PDBu on the calcium transients depends on PKC, a correlated study of the effect of phorbol esters on calcium transients and PKC activity was performed. The calcium transients and PKC activity were similarly affected by PDBu in ordinary and cold-accommodated frogs, but the effects occurred more quickly in the latter. However, they still changed in parallel as in ordinary frogs. 1 or 10 micromol/l, 4-alpha-phorbol, a PKC-inactive analogue of phorbol ester, caused a partial depression of the calcium transients in cold-accommodated frogs, while PKC activity was not affected. Moreover, the transient potentiation of the calcium transients induced by 1 micromol/l PDBu could be antagonized by the PKC inhibitors 10 micromol/l chelerythrine chloride or 10 micromol/l polymyxin B (PMB). All these results suggest that: (1) the transient potentiation of calcium transients induced by PDBu is caused by activation of PKC; (2) phorbol ester can depress the calcium transients by a mechanism that is independent of PKC. PMID- 10370089 TI - Characteristics of a transient outward current (sensitive to 4-aminopyridine) in Ca2+-tolerant myocytes isolated from the rabbit atrioventricular node. AB - A transient outward current (Ito) has been observed in the atrioventricular node (AVN), but its characteristics in Ca-tolerant AVN myocytes have not been investigated previously. In this study, Ito was measured from Ca-tolerant rabbit AVN myocytes at 37 degrees C, using the whole-cell patch-clamp technique. With interfering currents inhibited, 500-ms voltage-clamp pulses applied from -80 mV elicited Ito at potentials positive to -30 mV, which increased in magnitude with test potential amplitude. This current was completely blocked by external application of 5 mM 4-aminopyridine (4-AP). During a command pulse, Ito activated rapidly then inactivated with a bi-exponential time-course. Fast and slow time constants of current inactivation (tauf and taus, respectively) showed voltage dependence. At 0 mV, tauf was 14.5+/-2.7 ms and taus was 112.8+/-21. 2 ms, whilst at +60 mV tauf was 6.7+/-1.1 ms and taus was 63.7+/-9.2 ms (n=25). The steady state inactivation relationship showed half-maximal inactivation at -33.8 mV (n=8). Re-activation of Ito after an inactivating pre-pulse showed a bi exponential time-course of recovery: tau1 was 196+/-70 ms, and tau2 was 2707+/ 1010 ms (n=6, at -80 mV). Repetitive application of voltage-clamp test pulses showed that Ito inactivation accumulated on repetitive stimulation, but reached a steady state rapidly for a given pulse frequency (0. 2-1.0 Hz). AVN Ito was sensitive to the class 1 anti-arrhythmic flecainide (EC50 for peak current of 24 microM), which showed selectivity for the rapidly inactivating current component. Quinidine also inhibited Ito in a dose-dependent fashion, but did not affect the current time-course. Under voltage-clamp conditions, a simulated diastolic depolarisation from -70 to -45 mV did not significantly reduce Ito amplitude, and under current-clamp conditions 4-AP inhibited spontaneous action potentials. Although this is consistent with a significant role for Ito in shaping AVN activity, under the conditions of this study 4-AP also partially blocked the "rapid" delayed rectifier current, IKr, and so the effects of 4-AP on action potentials could not be attributed exclusively to its effects on Ito. PMID- 10370090 TI - Use of replication-deficient adenoviruses to study signal transduction pathways. Muscarinic responses in HSG and HT29 epithelial cell lines are mediated by G protein betagamma-subunits. AB - HSG and HT29 cells express muscarinic receptors that increase intracellular free Ca2+ ([Ca2+]i) by activating phospholipase Cbeta. In the present study, we have used the measurement of [Ca2+]i with Fura-2 to show that these receptors are of the M3 sub-type and that the increase in [Ca2+]i triggered when they are activated is not sensitive to pertussis toxin. We have also used replication deficient adenoviruses expressing wild-type and dominant-negative mutants of the alpha-subunits of the heterotrimeric G proteins, Gq and Gi2, to investigate the mechanisms by which these receptors control phospholipase Cbeta. We find that the Ca2+ response to 100 micromol/l carbachol is not affected by increased expression of the wild-type alpha-subunit of Gq, but is blocked by the dominant-negative mutant of Gq and by both the wild-type and the dominant-negative mutant alpha subunits of Gi2. Expression of alpha-subunits of Gi2 presumably blocks the response to carbachol by scavenging free betagamma-subunits. We conclude that in HSG and HT29 cells, the Ca2+ response to M3 receptor activation is mediated by the betagamma- rather than the alpha-subunits of Gq. PMID- 10370091 TI - Exogenous CCK and gastrin stimulate pancreatic exocrine secretion via CCK-A but also via CCK-B/gastrin receptors in the calf. AB - A predominance of the pancreatic cholecystokinin (CCK) receptor of the B/gastrin subtype (CCK-B/G) was reported in calves older than 1 month. Specific CCK-A and CCK-B/G receptor antagonists (SR 27897 and PD 135158, respectively) were used to identify the CCK receptor subtype involved in exogenous CCK- and gastrin-induced exocrine pancreatic responses. Conscious calves (2 months old) with catheterized pancreas, jugular vein and duodenum were used; the pancreatic juice was continuously reinfused. CCK (30 pmol kg-1 min-1, 40 min) evoked an increase in pancreatic juice flow and enzyme secretion, while the same dose of gastrin increased enzyme secretion alone. CCK-induced pancreatic secretion was abolished by SR 27897 (15 nmol kg-1 min-1, 55 min) and reduced by PD 135158 (0.15 nmol kg-1 min-1, 55 min). Gastrin-induced enzyme secretion was reduced by PD 135158 (50% to 90%) and to a lesser extent by SR 27897 (50% to 60%). These results demonstrate that CCK and gastrin in the physiological range stimulate pancreatic exocrine secretion in calves and that these effects are partly mediated by CCK-B/G receptors. Although CCK-A receptors are not predominantly expressed, they seem to play a major role in the response of pancreatic exocrine secretion to CCK. PMID- 10370092 TI - Localization of aquaporin-3 mRNA and protein along the gastrointestinal tract of Wistar rats. AB - Since specific proteins responsible for water transport (aquaporins, AQPs) have been identified in a great variety of tissues, we decided to study the presence of AQP3 in the gastrointestinal tract (GIT) of Wistar rats. Poly(A+) RNA was purified from the mucosa of the stomach, jejunum, ileum and colon, and gross detection of AQP3 mRNA was done by Northern blot analysis. In situ hybridization studies were carried out to precisely localize the distribution of this transcript. Sections of the different tissues were hybridized with @400-bp [35S]riboprobes. The results presented here demonstrate that AQP3 is expressed throughout the GIT, with its expression in the colon and ileum greater than that in the stomach. Immunohistochemistry experiments, using a polyclonal antibody against AQP3, revealed that AQP3 protein is present at the basolateral membrane of the epithelial cells lining the villus tip of the small intestine and colon. The finding of AQP3 in the intestinal epithelia strongly suggests that this protein functions as a pathway for water transport in this epithelium. PMID- 10370093 TI - A transient and a persistent calcium release are induced by chlorocresol in cultivated mouse myotubes. AB - The effect of 4-chloro-m-cresol (4-CmC), a stabilizing agent used in commercial preparations of the muscle relaxant succinylcholine, on intracellular free calcium levels in cultivated mouse myotubes was studied. Calcium signals were monitored with an inverted microscope equipped for fluorescence photometry using fura-2 as the calcium indicator. Upon bath application of 500 microM 4-CmC for 90 s, two separate calcium signals, a transient and a sustained one, could be regularly discriminated. First, with a delay of 2 s, the intracellular calcium concentration increased from 41+/-13 to 541+/-319 nM, peaked after 2-5 s and declined within 10 s to nearly resting values (n=36). Then, after a delay of up to 20 s, intracellular calcium rose quickly again to almost the same value and stayed elevated as long as the drug was applied. Upon drug removal, intracellular calcium rapidly decreased to a new level that was always slightly higher than the original base line. At 250 microM 4-CmC, the response was small, whereas at 500 microM it was at its maximum. Thus, the concentration-response curve was very steep. Replacement of extracellular calcium by EGTA and application of calcium channel blockers revealed that, for both the transient and the sustained response, calcium was released from intracellular stores. Pre-treatment with thapsigargin (0.1 microM) or ryanodine (10 microM) abolished both signal components. Repeated short-term applications of 4-CmC suggest that the two components may arise from different systems. PMID- 10370094 TI - Low edge damage container insert that adjusts intestinal forceps biopsies into Ussing chamber systems. AB - Ussing chamber experiments with human intestinal tissue are impeded by the small size of forceps biopsy specimens. Therefore, a miniaturized container insert featuring low edge damage was designed with an exposure area of only 0.05 cm2. It allows measurement of short-circuit current (ISC) and transmural resistance (Rt) on endoscopically obtained biopsy specimens, as well as alternating current impedance analysis and conductance scanning. Comparison with larger specimens mounted in a conventional Ussing chamber without the insert (exposure area 0.54 cm2) was made using rat jejunum and rectum. No differences in ISC, Rt, or secretory response were found, indicating proper sealing and prevention of edge damage, as well as tissue viability in the container system. If biopsy samples obtained from human rectum were mounted in the insert, the local resistance near the edge was almost the same as the overall resistance (52.3 Omega.cm2). Epithelial and subepithelial resistances of human rectum were 43+/-1 Omega.cm2 and 10+/-1 Omega.cm2, respectively. In conclusion, we present a tool that allows reliable Ussing-type, impedance, and conductance scanning measurements to be made from intestinal biopsy specimens. PMID- 10370095 TI - Cell-cycle-dependent changes in membrane potential of L-929 cells caused by the effect of hydrogen peroxide. AB - The changes in membrane potential of L-929 fibroblasts caused by H2O2 (10-50 microM) at different phases of the cell cycle were investigated. The membrane potential of cells in G0, early G1, and G2/M responded to H2O2 by hyperpolarizing, while cells in late G1/S responded by depolarizing. Quinidine (50 microM), a blocker of Ca2+-dependent K+ conductance, inhibited the hyperpolarization response of L-929 cells to H2O2 in all cases. Measurements of reactive oxygen species (ROS) production by the cells at the same cell cycle phases indicated that the hyperpolarization response of the cells is linked with minimal production of ROS, and that the depolarization response is associated with maximal production of ROS inside the cell. PMID- 10370097 TI - Effects of pH on spontaneous tension oscillation in skinned bovine cardiac muscle. AB - Skinned cardiac muscle preparations exhibit spontaneous tension oscillations (spontaneous oscillatory contractions; SPOCs) in the absence of Ca2+, and in the presence of MgATP, MgADP and inorganic phosphate (Pi; ADP-SPOC). Similar oscillations occur in the presence of sub-micromolar concentrations of Ca2+ under normal activating conditions without MgADP and Pi (Ca-SPOC). In the study presented here, we investigated the effects of pH on both types of SPOC in skinned bovine cardiac ventricular muscle. First, a decrease in pH increased the MgADP concentration required to induce the half-maximal isometric tension that is obtained in the absence of Ca2+ and in the presence of MgATP (ADP-contraction). The inhibitory effect of Pi on ADP-contractions was not affected by pH. Second, ADP-SPOCs occurred upon the addition of Pi to the solution that resulted in ADP contraction, and the relative amplitude and the period of the tension oscillation in the presence of 2 mM MgATP, 10 mM MgADP and 10 mM Pi were unchanged under all pH conditions examined (6.6, 7.0, 7.4). On the contrary, the relative amplitude and the period of the Ca-SPOCs were markedly diminished at pH 6.6. Finally, we constructed state diagrams showing the effects of pH on SPOC conditions. The state diagram shows that SPOCs occur less frequently under acidic conditions than at neutral pH. We suggest that the intermediate state of crossbridges that is required for SPOCs is more difficult to attain at a low pH. PMID- 10370096 TI - Regulation of L- and N-types of Ca2+ channels by intracellular ATP4- in frog dorsal root ganglion neurons. AB - The roles of free Mg2+ ions, ATP4- ions and Mg-ATP complexes in the regulation of N- and L-types of Ca2+ channels were studied in frog dorsal root ganglion (DRG) neurons using the whole-cell patch-clamp technique. Because Mg2+ ions interact with ATP4- ions to form Mg-ATP complexes, addition of one species can influence the concentrations of the other two. In this study their concentrations were carefully controlled by varying the concentrations of two constituents at a time while keeping the third constant. The effects of each of the three species on barium currents through L-type (IBaL) and N-type (IBaN) Ca2+ channels were plotted against its concentrations. The dose-response curves for ATP4- show that IBaL and IBaN proportionally increased with ATP4- concentrations up to 1 mM at three different Mg2+ concentrations. At a fixed concentration of ATP4-, IBaL and IBaN remained unchanged even when pMg changed from 3 to 5. Dose-response curves for IBaL and IBaN plotted against Mg-ATP concentration did not show a consistent pattern. H-7 and Mg2+ ions did not exert any blocking effect on the activity of either Ca2+ channel type, and neither dibutyryl-cAMP nor NKH-477 had any stimulating effect, suggesting that phosphorylation is not likely to be involved in ATP-induced potentiation. From these observations, it is concluded that L-type and N-type Ca2+ channels in frog DRG neurons are regulated by ATP4- ions alone, and that the neuronal Ca2+ channels are regulated by mechanisms that are different from those regulating the cardiac Ca2+ channels. PMID- 10370098 TI - Protein phosphotyrosine phosphatase inhibitors suppress regulatory volume decrease and the volume-sensitive Cl- conductance in mouse fibroblasts. AB - The effects of the protein tyrosine phosphatase (PTP) inhibitors, pervanadate, monoperoxo(picolinato)- oxo-vanadate(V) [mpV(pic)] and dephostatin, on regulatory volume decrease (RVD) and the volume-sensitive Cl- current in mouse L-fibroblasts were studied with the aid of video microscopy and the whole-cell patch-clamp technique. The RVD induced by the hyposmotic shift from 300 to 150 mosmol/l, was strongly suppressed in cells that had been pre-incubated in pervanadate (25 microM) or in mpV(pic) (10 microM), or subjected to extracellular application of dephostatin (20 microM). The acceleration in RVD caused by gramicidin (0.5 microM) was also slowed down by pervanadate pre-treatment, suggesting that the PTP inhibitors affected the volume-sensitive Cl- conductance. Inhibition of the volume-sensitive Cl- current by pervanadate (25 microM) pre-treatment and by acutely applied dephostatin (20 microM) was confirmed in the whole-cell experiments (by @70% and by @50%, respectively). Both pervanadate and dephostatin inhibited the outward and inward Cl- currents equally, which suggests that only the number of open channels was affected. The amplitude of the Cl- current decreased slowly during application of dephostatin and did not recover after its termination. We conclude that in mouse L-fibroblasts, similar to bovine chromaffin cells, inhibition of PTPs results in the suppression of both RVD and the volume-sensitive Cl- current. PMID- 10370099 TI - N-Terminal deletions of rKv1.4 channels affect the voltage dependence of channel availability. AB - Rat Kv1.4 potassium channels undergo rapid inactivation, which is mediated by the N-terminal structure of the polypeptide. This inactivation can be removed by N terminal deletion of about 20 residues. However, more substantial deletion (e.g. 37 residues) restores inactivation suggesting a second inactivating domain [Kondoh et al. J Biol Chem 272:19333-19338, 1997]. Here we provide evidence that this inactivation shares all properties with N-type inactivation. Pore mutations, which are supposed to affect C-type inactivation, have no effect. In addition, the redox regulation of inactivation, which is typical for Kv1.4 channels, can be conferred to the inactivation of the deleted constructs by incorporation of an N terminal cysteine residue. The most remarkable feature of this secondary inactivation is the existence of two components in the steady-state voltage dependence of inactivation. For mutant rKv1. 4Delta2-37 about 90% of the channels only activate when the holding membrane potential is more negative than about 120 mV; the remaining 10% show the typical threshold at -60 mV. Mutagenesis of the truncated channel affected the relative amplitudes of these two components, but not the voltage dependence. The results suggest that the secondary ball structures of rKv1.4 channels interact with the protein structures responsible for activation. PMID- 10370100 TI - Blockade of the delayed rectifier K+ currents, IKr, in rabbit sinoatrial node cells by external divalent cations. AB - We investigated the actions of various divalent cations on the delayed rectifier K+ currents (IKr) in rabbit sinoatrial node cells using the whole-cell voltage clamp technique in isotonic K+ solutions. External divalent cations decreased the amplitude of currents, accelerated the time course of deactivation and shifted the activation to positive potentials in a dose-dependent manner. The concentrations for half-maximum inhibition of the steady-state currents (KM) obtained at 0 mV were 0.63, 1.36, 1.65 and 2.16 mM for Ni2+, Co2+, Mn2+ and Ba2+, respectively. The effect was voltage dependent (KM decreased e-fold for 12.2-16.8 mV hyperpolarization), but the dependence did not vary significantly among different cations. Acceleration of the time course of current deactivation by the increase of divalent cation concentration was well fitted by the voltage dependent block model, and the binding rate constant (k1) was obtained. The binding rates for the ions took the following order: Ni2+ >Co2+ >Mn2+ >Ba2+. The degree of the shift of activation occurred in the same order: Ni2+ >Co2+ >Mn2+ >Ba2+. From these results, we conclude that IKr channels are non-selectively blocked by most divalent cations from the external side and that the binding site is located deep inside the channel, resulting in a steep voltage dependence of the blockade. PMID- 10370101 TI - The relative order of IP3 sensitivity of types 1 and 3 IP3 receptors is pH dependent. AB - The type-3 inositol 1,4,5-trisphosphate (IP3) receptor, in contrast to the type-1 IP3 receptor (IP3R), is not stimulated by sulfhydryl oxidation and is less sensitive to adenosine 5'-triphosphate. In the present study we compared the effect of pH on the Ca2+ release induced by IP3 and cytosolic Ca2+ between IP3R3 expressing 16HBE14o- cells and IP3R1-expressing A7r5 cells. Changing pH from 6.8 to 7.5 decreased the IP3 concentration required for half-maximal stimulation of IP3R3 (EC50) 10.7-fold (from 2.14 to 0.20 microM). Similar alkalinization decreased the IP3 concentration (EC50) for stimulation of IP3R1 only 2.5-fold (from 0.87 to 0.35 microM). IP3R1 is therefore the more sensitive isoform at pH 6.8, while IP3R3 is more sensitive at pH 7.5. Stimulation and inhibition of IP3R1 and -3 by low and high cytosolic [Ca2+] respectively was observed at both pH 6.8 and 7.5. Increasing [H+] shifted the Ca2+-activation curve of IP3R1 towards higher [Ca2+] but did not affect the Ca2+ dependence of IP3R3. We conclude that IP3R1 and -3 differ markedly in their response to protons. PMID- 10370102 TI - Lithium evokes a more pronounced natriuresis when administered orally than when given intravenously to salt-depleted rats. AB - The effects on renal sodium excretion of giving lithium chloride (LiCl; 0.75 mmol per kg body mass) by gavage or intravenously were investigated. The experiments were carried out on Wistar-Kyoto (WKY) or spontaneously hypertensive (SHR) rats in metabolic cages. The rats had been on a low-salt diet for 4 days. Urine excretion of water, sodium and potassium was followed before and for 24 h after giving LiCl. An oral dose of LiCl evoked a more pronounced renal sodium excretion in either strain of rat as compared to that following intravenous administration, in agreement with previous observations of the effects of giving sodium chloride. Choline chloride (1.5 mmol per kg body mass) given by gavage to WKY rats or SHR evoked no change in the renal excretion of sodium. Based on the results of the present study and on observations reported in the literature, we propose that the intestinal tract contains a sodium "sensor", which upon activation releases a natriuretic factor to cause renal sodium excretion. The present results indicate that the proposed "sensor" is sensitive to lithium but not chloride ions. PMID- 10370103 TI - The Na+2Cl-K+ cotransporter in the rectal gland of Squalus acanthias is activated by cell shrinkage. AB - Effects of cAMP on Cl- secretion, intracellular Cl- activity and cell volume were studied in isolated perfused rectal gland tubules (RGT) of Squalus acanthias with electrophysiological and fluorescence methods. Recording of equivalent short circuit current (Isc) showed that cAMP stimulates Na+Cl- secretion in a biphasic manner. The first and rapid phase corresponds to Cl- exit via the respective protein-kinase-A- (PKA-) phosphorylated Cl- conductance. The inhibitory effect of the loop diuretic furosemide (0.5 mmol/l, n=12) indicates that second phase reflects the delayed (1-2 min) activation of the Na+2Cl-K+ cotransporter. During the first phase cytosolic Cl- activity, as monitored by 6-methoxy-N-(3 sulfopropyl) quinolinium (SPQ) fluorescence, fell to 78% (n=23) of the control value. Concomitantly, a transient fall in cell volume was recorded by calcein fluorescence to 92% (n=5) of the control value. Preincubation of the RGT with phalloidin (0.1 mmol/l, n=6) or cytochalasin D (0.1 mmol/l, n=4) almost completely prevented the development of the second phase of Isc activation. When cytosolic Cl- activity was increased by exposing the RGT to a high K+ concentration (25 mmol/l), in the presence of mannitol to prevent volume increases, stimulation was unaffected and biphasic. In contrast, when cell volume was clamped to an increased value (115%, n=8) by removing extracellular NaCl, the second phase was abolished completely (n=11). These data suggest that the primary and key process for triggering the Na+2Cl-K+ cotransport is transient cell shrinkage. PMID- 10370104 TI - Cholinergic modulation of non-adrenergic, non-cholinergic relaxation in isolated, small coronary arteries from lambs. AB - The presence of cholinergic innervation of small coronary arteries in the lamb was investigated by measuring choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) activities and by performing in vitro experiments in a microvascular myograph to establish whether or not there is a cholinergic component in the response to electrical field stimulation (EFS). ChAT-specific activity was present in proximal coronary segments, but was significantly higher in small coronary arteries. AChE-positive ganglia and fibres were distributed within the adventitia and outer third of the media in proximal coronary segments, and dense perivascular nerve plexuses were observed in small coronary arteries. Acetylcholine induced contractions in all preparations examined and relaxations in 20% of the segments contracted with the thromboxane analogue U46619. EFS did not induce neurogenic contractions in lamb small coronary arteries. In the presence of the alpha-adrenoceptor antagonist, phentolamine, EFS caused frequency dependent reproducible relaxations that were enhanced by the blocker of cholinergic transmission, botulinum neurotoxin. An inhibitor of AChE, physostigmine, had no significant effect on the relaxations caused by EFS, while both the muscarinic receptor antagonist, atropine, and the muscarinic M2-receptor antagonist, AFDX 116, enhanced these responses. Blockade of sympathetic neurotransmission with guanethidine or incubation with the P2-receptor antagonist, suramin, abolished the relaxations induced by EFS, whereas propranolol was without effect. Low-frequency EFS caused less relaxation in preparations activated by acetylcholine than in those contracted with U46619, while sensitivity and maximal relaxation induced by adenosine 5'-triphosphate (ATP) were not different in U46619- and acetylcholine-contracted arteries. The presence of the enzymes necessary for both biosynthesis and degradation of acetylcholine and the finding that blockers of cholinergic neurotransmission enhance EFS-induced relaxations suggest that small coronary arteries are cholinergically innervated. PMID- 10370105 TI - The properties of the inward rectifier potassium currents in rabbit coronary arterial smooth muscle cells. AB - In freshly-isolated, single, smooth muscle cells of rabbit coronary arteries, an inward rectifier K+ current [IK(IR)] was identified using the whole-cell voltage clamp technique. The current/voltage (I/V) relationship of IK(IR) showed strong inward rectification with a very small outward current when the smooth muscle cells were dialyzed with a pipette solution containing Mg2+. However, dialyzing the cells with a nominally Mg2+-free pipette solution revealed a significant outward current hump in the I/V relation of IK(IR), suggesting that the strong inward rectification of IK(IR) is partly due to the inhibitory effects of internal Mg2+. IK(IR) was unaffected by tetraethylammonium (1 mM), 4 aminopyridine (1 mM), or glibenclamide (1 microM), but was inhibited by extracellular Ba2+ with a concentration of 0.87 microM eliciting half-maximal inhibition at -120 mV. IK(IR) induced in rabbit coronary smooth muscle cells declined during very negative hyperpolarizing steps, due largely to a block by external Na+. IK(IR) was inhibited by alpha1-adrenergic stimuli. Methoxamine, an alpha1-adrenergic agonist, concentration dependently inhibited IK(IR) in the presence of the beta-adrenergic antagonist propranolol. The methoxamine concentration required for half-maximal inhibition was 205 microM. We conclude that inward rectifier K+ current is present in rabbit coronary smooth muscle cells and that it shares many properties with the inward rectifier K+ current described for other cell types. PMID- 10370106 TI - Forskolin increases apical sodium conductance in cultured toad kidney cells (A6) by stimulating membrane insertion. AB - The role of membrane traffic in the stimulation of apical Na+ permeability caused by increases in cytoplasmic cyclic AMP was assessed by measuring the effects of forskolin on transepithelial capacitance (CT), transepithelial conductance (GT), and short-circuit current (Isc) in A6 cultured toad kidney cells. Apical water permeability was probed by recording cell volume changes after reducing the osmolality of the apical bath. We found that forskolin does not increase the osmotic water permeability of the apical membrane of A6 cells, and thus does not stimulate the insertion of water channels. Comparison of the effects of forskolin and insulin on Na+ transport demonstrated that both agents produce reversible increases in CT, GT and Isc. GT and CT increased proportionally during the rising phase of the insulin response. However, a non-linear relationship between both parameters was recorded when forskolin was given in NaCl Ringer's solution. The relationship between CT and GT became linear after the effects of forskolin on Cl conductances were eliminated by substituting Cl- by an impermeant anion. In contrast, in Cl--containing Na+-free solutions, the non-linearity became more pronounced. Successive additions of insulin and forskolin caused additive increases in CT. Because increases in CT and Na+ transport occurred in the absence of stimulation of water permeability and increases of CT and GT were directly proportional when Na+ was the major permeating ion across the apical membrane, we suggest that the increase in apical Na+ permeability in the presence of either forskolin or insulin is due to the insertion of channels residing in intracellular pools. In contrast, the increased Cl- permeability caused by forskolin may be related to the activation of channels already present in the membrane. PMID- 10370107 TI - Corticotropin-releasing hormone acts on guinea pig ileal smooth muscle via protein kinase A. AB - In contraction studies corticotropin-releasing hormone (CRH) was found to relax ileal but not gastric and jejunal smooth muscles of the guinea-pig, precontracted with BaCl2. Under whole-cell patch-clamp conditions, CRH concentration dependently activated Ca2+-sensitive K+ currents (IK) with ED50=20 pM at 100 nM and ED50=0. 13 pM at 500 nM intracellular Ca2+ respectively. This increase was accompanied by significant hyperpolarization of the cell membranes. CRH 9-41 peptide fragment did not affect IK amplitude, membrane potential or contraction. The CRH-induced increase of IK densities was accelerated in the presence of high intracellular Ca2+ concentrations (500 nM) and was abolished by pretreatment of cells with either ryanodine or thapsigargin, which cause depletion of intracellular Ca2+ stores, as well as in cells treated under conditions prohibiting intracellular Ca2+ store refilling. The effect of CRH on IK was not affected by bath application of various selective inhibitors of membrane-bound phospholipases, protein kinase C, cGMP-dependent protein kinase or Ca2+/calmodulin-dependent protein kinase II, but was effectively antagonized by blockers of protein kinase A (PKA) or adenylyl cyclase. Neither forskolin nor the catalytic subunit of PKA could mimic the effect of CRH on IK. Thus, it was suggested that CRH exerts its relaxing activity on ileal smooth muscle cells via PKA-dependent phosphorylation of some intracellular target coupled to sarcoplasmic reticulum Ca2+ storage machinery. PMID- 10370108 TI - A global defect in scaling relationship between electrical activity and availability of muscle sodium channels in hyperkalemic periodic paralysis. AB - Hyperkalemic periodic paralysis (HyperPP) is a hereditary disorder characterized by alternate episodic attacks of muscle weakness and muscle myotonia. The most common mutation associated with HyperPP is a T704M substitution in the skeletal muscle sodium channel. This mutation increases sodium persistent currents, alters voltage dependence of activation and impairs slow inactivation. The present study shows experimental evidence in support of a potentially important global defect caused by the T704M mutation. While the effective rate of recovery from slow inactivation, in both normal and mutated channels, is related to the duration of past activity by a power law function, the scaling power of the mutated channel is significantly greater. This difference between the channels offers a clue for an explanation to the wide range of time scales, history dependence, and the mixed myotonic/paralysis effect, which mark the clinical picture of HyperPP. PMID- 10370109 TI - The role of tumour necrosis factor-alpha (TNF-alpha) in fever and the acute phase reaction in rabbits. AB - We investigated the role of tumour necrosis factor-alpha (TNF-alpha) in fever and the acute phase reaction using a specific type-IV phosphodiesterase inhibitor, rolipram, that inhibits the production of TNF-alpha. The body temperatures and serum iron concentrations of rabbits were measured following injections of lipopolysaccharide (LPS) or Staphylococcus aureus (S. aureus) with either rolipram, diclofenac sodium or the appropriate control solutions. Rolipram significantly (P<0.05) inhibited the first phase of both LPS and Staphylococcal fever, but had no effect on the second phase. The fall in serum iron concentration was not significantly affected by the injection of rolipram together with LPS or S. aureus. These results suggest that TNF-alpha is a pyrogen that plays a role during the first phase of fever, at least. However, TNF-alpha appears not to mediate the fall in serum iron concentration during the acute phase reaction. PMID- 10370110 TI - Inhibition of force and shortening in smooth muscle by vanadate. AB - We have investigated the effects of vanadate (Vi) on force generation by, and shortening of, chemically skinned smooth muscle preparations from guinea-pig taenia coli at 22 degrees C. A method, using phosphatase inhibitors, was introduced to obtain stable, long-lasting contractions in thiophosphorylated preparations. Vi (10-1000 microM) dose-dependently inhibited active force, to about 20% of its maximum level. At a higher temperature (30 degrees C), the rate of inhibition was faster but the extent of inhibition was less. The rate of contraction following photolytic release of ATP to fibres in rigor was not affected by Vi (30 microM). The maximal shortening velocity (Vmax) was inhibited in a similar manner as active force by Vi (30 microM). In conclusion, the results suggest that Vi interacts with a force-generating actomyosin-ADP (AMADP) state reached after phosphate release. The rate of inhibition of smooth muscle contraction was markedly lower than in skeletal muscle, suggesting differences either in properties of the Vi-bound states or, more likely, in the concentration of AMADP states capable of binding Vi. This suggests that the long duty cycle in smooth muscle is not associated with a higher relative population of AMADP states reached immediately after Pi release, but rather by an increase in the population of subsequent force-generating cross-bridge states. The Vi-bound cross-bridges introduce an internal load to shortening, possibly acting in a similar manner as cross-bridge states introduced at low levels of activation. PMID- 10370111 TI - Measurement of concentration-response relationships by concentration-ramp application of agonists. AB - Dose-response relationships of agonist- induced membrane currents were obtained during accumulation of agonist and a simultaneously applied dye in the culture dish. The concentration of the dye was continuously measured using a laboratory made photometer. When the superfusate was switched from the control solution to a test solution containing an agonist and an inert dye (e.g. Fast Green) both accumulated in the bath together and the change of dye concentration altered the intensity of light falling through the objective. The change in light intensity in an area directly surrounding the cell under study was measured by the photometer. The normal microscope lamp was used as the light source and a colour filter was placed between the microscope lamp and the condenser in order to enhance the signal-to-noise ratio of the photometer output. Assuming that the rates of accumulation and clearance of dye are similar to those of the agonist, it is possible to continuously measure the membrane current at given dye and agonist concentrations. To calculate the dose-response curves we calibrated the relationship between the dye concentration and the photometer voltage output. PMID- 10370112 TI - Tachycardiac response in SHR and WKY rats induced by stress. PMID- 10370113 TI - Hox C6 expression during development and regeneration of forelimbs in larval Notophthalmus viridescens. AB - A central theme concerning the epimorphic regenerative potential of urodele amphibian appendages is that limb regeneration in the adult parallels larval limb development. Results of previous research have led to the suggestion that homeobox containing genes are "re-expressed" during the epimorphic regeneration of forelimbs of adult Notophthalmus viridescens in patterns which retrace larval limb development. However, to date no literature exists concerning expression patterns of any homeobox containing genes during larval development of this species. The lack of such information has been a hindrance in exploring the similarities as well as differences which exist between limb regeneration in adults and limb development in larvae. Here we report the first such results of the localization of Hox C6 (formerly, NvHBox-1) in developing and regenerating forelimbs of N. viridescens larvae as demonstrated by whole-mount in situ hybridization. Inasmuch as the pattern of Hox C6 expression is similar in developing forelimb buds of larvae and epimorphically regenerating forelimb blastemata of both adults and larvae, our results support the paradigm that epimorphic regeneration in adult newts parallels larval forelimb development. However, in contrast with observations which document the presence of Hox C6 in both intact, as well as regenerating hindlimbs and tails of adult newts, our results reveal no such Hox C6 expression during larval development of hindlimbs or the tail. As such, our findings indicate that critical differences in larval hindlimb and tail development versus adult expression patterns of this gene in these two appendages may be due primarily to differences in gene regulation as opposed to gene function. Thus, the apparent ability of urodeles to regulate genes in such a highly co-ordinated fashion so as to replace lost, differentiated, appendicular structures in adult animals may assist, at least in part, in better elucidating the phenomenon of epimorphic regeneration. PMID- 10370114 TI - Incorporation of mouse zona pellucida proteins into the envelope of Xenopus laevis oocytes. AB - All vertebrate eggs have extracellular matrices, referred to as the zona pellucida in Mus musculus and the vitelline envelope in Xenopus laevis. The mouse zona, composed of three sulfated glycoproteins (ZP1, ZP2, ZP3), is critical for fertilization and early development, and mice lacking a zona pellucida produce no live offspring. The primary structures of mouse ZP1 (623 amino acids), ZP2 (713 amino acids) and ZP3 (424 amino acids) have been deduced from full-length cDNAs, but posttranslational modifications result in mature zona proteins with molecular masses of 200-180 kDa, 140-120 kDa, and 83 kDa, respectively. The vitelline envelope forms a similar structure around Xenopus eggs and contains three glycoproteins that are structurally related (39-48% amino acid similarity) to the three mouse zona proteins. To investigate whether the structural semblances are sufficient to allow incorporation of the mouse zona proteins into the Xenopus vitelline envelope, capped synthetic mRNAs encoding ZP1, ZP2, and ZP3 proteins were injected into the cytoplasm of stage VI Xenopus oocytes. After 20 h of incubation the oocytes were harvested, and posttranslationally modified zona proteins were detected with monoclonal antibodies specific to mouse ZP1, ZP2, and ZP3. The oocytes were imaged with confocal microscopy to detect individual zona proteins in the extracellular matrix of the oocytes, and this localization was confirmed biochemically. Thus the mouse zona proteins appear to have been sufficiently conserved through 350 million years of evolution to be incorporated into the extracellular envelope surrounding Xenopus eggs. PMID- 10370115 TI - The role of wingless in the development of multibranched crustacean limbs. AB - Arthropods are the most diverse and speciose group of organisms on earth. A key feature in their successful radiation is the ease with which various appendages become readily adapted to new functions in novel environments. Arthropod limbs differ radically in form and function, from unbranched walking legs to multibranched swimming paddles. To uncover the developmental and genetic mechanisms underlying this diversification in form, we ask whether a three-signal model of limb growth based on Drosophila experiments is used in the development of arthropod limbs with variant shape. We cloned a Wnt-1 ortholog (Tlwnt-1) from Triops longicaudatus, a basal crustacean with a multibranched limb. We examined the mRNA in situ hybridization pattern during larval development to determine whether changes in wg expression are correlated with innovation in limb form. During larval growth and segmentation Tlwnt-1 is expressed in a segmentally reiterated pattern in the trunk. Unexpectedly, this pattern is restricted to the ventral portion of the epidermis. During early limb formation the single continuous stripe of Tlwnt-1 expression in each segment becomes ventrolaterally restricted into a series of shorter stripes. Some but not all of these shorter stripes correspond to what becomes the ventral side of a developing limb branch. We conclude that the Drosophila model of limb development cannot explain all types of arthropod proximodistal outgrowths, and that the multibranched limb of Triops develops from an early reorganization of the ventral body wall. In Triops, Tlwnt-1 plays a semiconservative role similar to that played by Drosophila wingless in segmentation and limb formation, and morphological innovation in limb form arises in part through an early modulation in the expression of the Tlwnt-1 gene. PMID- 10370116 TI - Genetic analysis of steel and the PG-M/versican-encoding gene AxPG as candidates for the white (d) pigmentation mutant in the salamander Ambystoma mexicanum. AB - Vertebrate non-retinal pigment cells are derived from neural crest (NC) cells, and several mutations have been identified in the Mexican axolotl Ambystoma mexicanum (Ambystomatidae) that affect the development of these cell lineages. In "white" (d) mutant axolotls, premigratory NC cells differentiate as pigment cells, yet fail to disperse, survive, or both, and this leads to a nearly complete absence of pigment cells in the skin. Previous studies revealed that d affects pigment cell development non-autonomously, and have reported differences between white and wild-type axolotls in the structure and composition of the extracellular matrix through which NC and pigment cells migrate. Here we test the correspondence of d and two candidate genes: steel and AxPG. In amniotes, Steel encodes the cytokine Steel factor (mast cell growth factor; stem cell factor; kit ligand), which is expressed along the migratory pathways of melanocyte precursors and is required by these cells for their migration and survival; mammalian Steel mutants resemble white mutant axolotls in having a deficit or complete absence of pigment cells. In contrast, AxPG encodes a PG-M/versican-like proteoglycan that may promote the migration of A. mexicanum pigment cells, and AxPG expression is reduced in white mutant axolotls. We cloned a salamander orthologue of steel and used a partial genetic linkage map of Ambystoma to determine the genomic locations of steel, AxPG, and d. We show that the three genes map to different linkage groups, excluding steel and AxPG as candidates for d. PMID- 10370117 TI - sox30: a novel zebrafish sox gene expressed in a restricted manner at the midbrain-hindbrain boundary during neurogenesis. AB - The Sox family of proteins is thought to act to regulate gene expression in a wide variety of developmental processes. Here we describe the cloning of sox30, a novel sox gene from the zebrafish (Danio rerio). In situ hybridization shows that sox30 is expressed in a restricted manner at the boundary between the midbrain and hindbrain during nervous system development. This expression pattern is in direct contrast to that of most other neuronally expressed Sox genes which are expressed throughout the nervous system. PMID- 10370118 TI - Neurogenesis during caudal spinal cord regeneration in adult newts. AB - After tail amputation in urodele amphibians, dramatic changes appear in the spinal cord rostral to the amputation level. Transection induces a proliferation response in cells lining the ependymal canal, giving rise to an ependymal tube in which neurogenesis occurs. Using the thymidine analog bromodeoxyuridine (BrdU) in short- and long-term labeling of cells undergoing DNA synthesis (S phase of the cell cycle), specific cell markers, and cell cultures, we show that neurons derive from the proliferative ependymal layer of the ependymal tube. PMID- 10370119 TI - Distinct expression patterns of different enhancer of split bHLH genes during embryogenesis of Drosophila melanogaster. AB - E(spl) bHLH genes are targets of the Notch pathway: they are transcriptionally activated in response to the Notch signal. Yet, during imaginal development, additional regulatory factors appear to modulate transcription resulting in different expression patterns. During early embryogenesis all E(spl) bHLH genes are expressed in roughly the same domain, namely the neurogenic ectoderm. Within this region these seven genes show a highly dynamic, yet distinct transcriptional activity. Our analysis further detected tissue specific expression of some E(spl) genes at later embryonic stages. Prominent differences were observed in the dorsolateral and procephalic neuroectodermal regions as well as in the mesoderm. These observations indicate that other factors in addition to the Notch signal participate in the regulation of the individual E(spl) genes not only in imaginal tissues but also during neuroblast specification and other cell fate determination events in the embryo. PMID- 10370120 TI - The MADS domain containing transcription factor cMef2a is expressed in heart and skeletal muscle during embryonic chick development. AB - Muscle enhancer factor 2 (MEF2) proteins are important transcription factors for muscle-specific gene activation. Four family members are known in mammals, referred to as MEF2A, MEF2B, MEF2C, and MEF2D. Here we report the isolation and expression pattern of the chick Mef2a gene (cMef2a). cMef2a expression starts in precardiac mesoderm of HH stage 8 embryos. During further embryonic development expression continues in the heart tube and later in atrium and ventricle. A second cMef2a expression domain appears in somites of stage 13 embryos. Somitic cMef2a expression is limited to the myotome and is not found in newly formed somites until the muscle-specific transcription factors MyoD and myogenin are present. This suggests that activation of the cMef2a gene in skeletal muscle is dependent on these basic helix-loop-helix transcription factors. cMef2a expression in heart and skeletal muscle continues into adulthood when it is also seen in intestinal mesenchyme and in brain. PMID- 10370121 TI - A system to efficiently maintain embryonic lethal mutations in the flour beetle Tribolium castaneum. AB - Due to its small size, short life cycle, and easy maintenance, the flour beetle Tribolium castaneum is well suited for the genetic analysis of development. One drawback of Tribolium as a genetic system is, however, the difficulty of keeping embryonic lethal lines. Presently, only few lethal mutations can be kept as balanced stocks. Therefore, heterozygous carriers must be identified anew in every generation in order to maintain a recessive embryonic mutation. To alleviate this problem we have devised a block system that allows the simultaneous processing of many mutant lines or test crosses for visual inspection of larval cuticle phenotypes. Using this technique, one person can maintain about 100 embryonic lethal stocks, which makes feasible the thorough genetic analysis of embryogenesis in this species. PMID- 10370122 TI - Characterization of the Tribolium Deformed ortholog and its ability to directly regulate Deformed target genes in the rescue of a Drosophila Deformed null mutant. AB - We have analyzed the Tribolium castaneum ortholog of the Drosophila homeotic gene Deformed (Dfd) and determined its expression pattern during embryogenesis in this beetle. Tc Deformed (Tc Dfd) is expressed in the blastoderm and the condensing germ rudiment in a region that gives rise to gnathal segments. During germ band extension Tc Dfd is expressed in the mandibular and maxillary segments, their appendages, and the dorsal ridge. Comparison of insect Dfd protein sequences reveals several highly conserved regions. To determine whether common molecular features reflect conserved regulatory functions we used the Gal4 system to express the Tribolium protein in Drosophila embryos. When Tc Dfd is expressed throughout embryonic ectoderm under the control of P69B, the beetle protein autoregulates the endogenous Dfd gene. In addition, the Drosophila proboscipedia gene (a normal target of Dfd) is ectopically activated in the antennal and thoracic segments. We also compared the ability of the beetle and fly proteins to rescue defects in Dfd- mutants by expressing each throughout the embryonic during embryogenesis. Both proteins rescued Dfd- defects to the same extent in that they each restore the development of mouth hooks and cirri, as well as cause gain-of function abnormalities of posterior mouth parts. As before, pb was ectopically activated in the antennal segment. This is the first demonstration of the ability of a heterologous homeotic selector protein to directly regulate a target gene independent of an endogenous Drosophila autoregulatory loop. PMID- 10370123 TI - The zebrafish eya1 gene and its expression pattern during embryogenesis. AB - The eyes absent-like genes encode a group of putative transcriptional coactivators with a sole representative in Drosophila and several members in mammals. Haploinsufficiency of the human EYA1 gene results in branchio-oto-renal syndrome characterized by developmental anomalies of the branchial arches, the three compartments of the ear and the kidney. As a first step towards a functional analysis of this gene in lower vertebrates, we isolated its zebrafish homologue, eya1, and studied its expression pattern during embryogenesis. The eya1 cDNA predicts a protein with 84.7% identity with the human homologue. Transcripts are first detected at the tailbud stage in presumptive cranial placodal precursor cells. Thereafter, eya1 expression continues in anterior pituitary, olfactory, otic, and lateral line placodes. Aside from these placodal sites of expression, eya1 transcripts were observed in the somites, developing pectoral fins, and branchial arches. No expression was found in pronephros or Wolffian duct of the zebrafish renal system. Within the developing ear, eya1 expression becomes confined to the ventral part of the otic vesicle from where the acoustic ganglion precursor cells arise and the sensory patches differentiate. In the lateral line, eya1 is expressed in the placodes, ganglia, migrating primordia, and receptive organs at all developmental stages, including both the differentiating hair and supporting cells. Taken together, these results indicate a remarkable similarity in both the structure and expression pattern of eya1 between higher and lower vertebrates, suggesting that the function of this gene has been conserved throughout vertebrate evolution. PMID- 10370124 TI - Expression of MADS box genes ZMM8 and ZMM14 during inflorescence development of Zea mays discriminates between the upper and the lower floret of each spikelet. AB - Most floral meristem and organ identity genes of dicotyledonous plants belong to the MADS box gene family. Since they are generally transcribed in those tissues and organs whose identity they determine, they are excellent markers for developmental processes. Here we report the cDNA cloning of a pair of MADS box genes, ZMM8 and ZMM14, from the monocotyledonous plant maize. Maize inflorescences are composed of spikelets which contain two florets, an upper and a lower one. Although upper and lower florets develop in a very similar way in male inflorescences, ZMM8 and ZMM14 expression was found in all organs of upper florets, but no transcripts were detected in lower florets. In contrast, two other MADS box genes were found to be expressed in lower florets in the same way as in upper florets. Our observations suggest that during spikelet development ZMM8 and ZMM14 work as selector genes which are involved in distinguishing the upper from the lower floret. Alternatively, these genes may be involved in conferring determinacy to the spikelet or upper floret meristem. Our data suggest that in the phylogenetic lineage that led to maize an ancient type of MADS box gene has been recruited during evolution for the establishment of novel positional information not found within the simple inflorescences of dicotyledonous plants such as Arabidopsis. PMID- 10370125 TI - Ectopic mesodermal expression promoted by the eight-cell stage Bufo arenarum subequatorial cytoplasm. AB - Mesodermal determinants were investigated by cytoplasmic transfer and blastomere isolation in the eight-cell stage of Bufo arenarum. Their existence was confirmed by assaying the subequatorial cytoplasm's ability to respecify the developmental potency of animal quartets. The gray subequatorial cytoplasm, but not animal cytoplasm, is able to divert the ectodermal fate of animal quartets to several mesodermal components. The source of the transplanted cytoplasm was important in determining the category of the resulting structures. Ventral subequatorial cytoplasm from ventrovegetal blastomeres generated ventral derivatives, namely erythrocytes and mesenchyma. Dorsal subequatorial cytoplasm from dorsovegetal blastomeres produced dorsolateral derivatives, such as notochord, muscle, nephric tubules, and coelomic epithelium, including mesenchyma. On the other hand, transfer of vegetal pole cytoplasm to animal quartets resulted in the formation of groups of endoderm-like cells dispersed among epidermal cells. However, the presence of such cells did not cause any mesodermal induction. The present findings suggest the existence of cytoplasmic information responsible for mesodermal specification. The alternative hypothesis that animal blastomeres become mesoderm due to vegetal induction is questioned. PMID- 10370127 TI - Differential expression of two nonallelic MyoD genes in developing and adult myotomal musculature of the trout (Oncorhynchus mykiss). AB - Previously we identified two nonallelic MyoD encoding genes in the rainbow trout. These two MyoD genes (TMyoD and TMyoD2) were duplicated during the tetraploidization of the salmonid genome. In this study we show that TMyoD and TMyoD2 exhibit a distinct spatiotemporal pattern of expression that defines discrete cell populations in the developing somite. TMyoD expression is first detected in the mid-gastrula on either side of the elongating embryonic shield. During the anterior-to-posterior wave of somite formation the TMyoD transcript is initially present in adaxial cells of both the presomitic mesoderm and the forming somites. A lateral extension of TMyoD expression occurs only when the myotomes acquire their characteristic chevron shape pointing rostrally. By contrast, the initial expression of TMyoD2 occurs in somites that have already formed and is limited to the posterior compartment of somites. Further, in postlarval trout we observed a differential expression of TMyoD and TMyoD2 genes in muscle fibers with differing phenotype. Collectively, these data provide evidence that the two trout MyoD encoding genes have evolved to become functionally different. A comparison of the expression patterns of the two trout MyoD genes with that of myogenin allowed us to position them in the regulatory pathway leading to muscle differentiation. PMID- 10370126 TI - Tissue-specific requirements for a phosphorylation site in the Fushi tarazu homeodomain. AB - The homeodomain protein Fushi tarazu (Ftz) is required for several embryonic patterning processes including segmentation and neurogenesis. During the stages that these processes are regulated the protein is differentially phosphorylated, suggesting that phosphorylation plays a role in helping the protein to regulate different functions in different tissues. We showed in a recent study that one of the Ftz phosphorylation sites, a protein kinase A-type site in the N-terminal arm of the homeodomain, is required for normal Ftz-dependent segmentation. Here we test whether phosphorylation of this site (Thr-263) is also required in the developing central nervous system (CNS). A well-established role for Ftz in the CNS is for the differentiation of neurons referred to as RP2 neurons. Absence of Ftz expression in these cells causes a failure of certain target genes to be expressed and subsequent defects in RP2 differentiation. In contrast to its effect on segmentation, we find that mutation of Thr-263 to Ala (or Asp) has no effect on these CNS functions. This suggests that the phosphorylation state of this site is irrelevant for Ftz function in the CNS, and that there are tissue specific differences in the requirements for Ftz phosphorylation. PMID- 10370128 TI - Differential response of fetal and neonatal myoblasts to topographical guidance cues in vitro. AB - Fusion of mononucleated myoblasts into parallel arrays of mutinucleated myotubes is an essential step in skeletal myogenesis. The formation of such a highly ordered structure requires myoblasts to come together, orient and align in the correct location prior to fusion. We report here that fetal and neonatal myoblasts can use topographical features as strong guidance cues in vitro. Myoblasts were cultured on multiple grooved substrata of varying dimensions, and the axial orientations of individual cells were recorded. Both fetal and neonatal myoblasts aligned parallel with the direction of deep grooves (2.3-6.0 micron), which is correlated well with the location of myoblasts in similar sized grooves during secondary myogenesis. Fetal myoblasts also responded to shallower grooves (0. 04-0.14 micron) by aligning parallel or perpendicular to the direction of the grooves, indicating the ability of these cells to respond to fine elements normally encountered within the developing muscle architecture. In contrast, neonatal myoblasts failed to respond to shallow grooves, adding to the suggestion that fetal and neonatal myoblasts may represent separate populations of myoblasts. Overall, the results demonstrate that myoblasts respond to large and small features of the physical topography in vitro and indicate that structural elements in the microenvironment of the muscle may play a critical role in myoblast spatial organization during myogenesis. PMID- 10370129 TI - Zebrafish CTH1, a C3H zinc finger protein, is expressed in ovarian oocytes and embryos. AB - The Zfcth1 gene is, as the previously cloned carp cth1 gene, related to the mammalian TIS 11 family of primary response genes and encodes a protein with two putative CCCH zinc fingers. This report describes the RNA expression of this gene during oogenesis and early embryogenesis up to gastrulation in the zebrafish (Danio rerio). Maternal cth1 message is present in the ovary of 1-month-old fish and of adult fish in oocytes at all stages of maturation. In the youngest oocytes the message is localized in the cytoplasm all around the nucleus, in larger oocytes the message becomes restricted to the future animal pole of the embryo, and in mature oocytes the expression is sharply localized in the cortical layer under the micropyle. After ovulation the cth1 messenger spreads over the cytoplasmic cap and is distributed over the blastomeres during subsequent cleavages. In subsequent stages maternal expression of cth1 gradually disappears. From early epiboly stages onward embryonic cth1 expression is localized to the germ ring and the hypoblast cells in the central part of the embryonic shield. In the shield, cth1 expression largely overlaps with the area of gooscoid expression in the first involuting cells. In stages after 70% of epiboly cth1 expression diminishes and soon can no longer be detected in the embryo. Next to a developmental role in cell fate determination we propose a function for cth1 during oocyte maturation. PMID- 10370130 TI - Gene transfer into insect brain and cell-specific expression of bombyxin gene. AB - A transgene reporter consisting of the bombyxin gene promoter and the green fluorescent protein coding region was introduced into intact brains of the silkworm Bombyx mori by in vitro electroporation. After in vitro culture of the brains, the fluorescence derived from the introduced reporter gene was observed in all cases in eight neurosecretory cells that had previously been identified as bombyxin-producing cells (BPCs). Although the fluorescence was not always observed in all cells, it was specific to BPCs, indicating that the reporter was under the control of the bombyxin gene promoter in a BPC-specific manner. Electroporatical introduction of a reporter gene was therefore found to be a suitable method for analyzing cell-specific expression in intact tissues and to be substitute for germ-line transmission of reporters in the transgenic system. Application of this technique enables us to analyze the cell-specific expression of transgene reporters within a few days and treat more than several dozens of the reporters within 1 month, which is difficult to do with the transgenic system. PMID- 10370131 TI - The application of thermodynamic principles to histochemical and morphometric tissue research: principles and practical outline with focus on the glycosciences. AB - Physicochemical terms such as entropy or current of entropy are commonly used to refer solely to the description of reactions in the realm of chemistry and physics. Since these thermodynamic terms have a predictive value for the further course of development of such reactions, e.g., extent of a chemical reaction or affinity of molecular interactions, it is tempting to introduce the respective algorithms to biological problems. By combining quantitative morphology with the histochemical visualization of distinct cellular and textural properties such as nuclear DNA contents or intensity of histochemical staining, equations from the general theory of thermodynamics can be adapted. They permit appropriate calculations to be performed which introduce the entropy concept to the processing of the information collected by analysis of structures formed by histochemically labeling cells. The theory of weighted graphs offers the appropriate mathematical tools. Nuclei are defined as vertices. Their DNA contents measured by the integrated optical density or additional cellular features (for example staining intensity of applied immuno-/ligandohistochemical probes) define the associated weights and the minimum spanning tree as a derivative from Voronoi's theorem for the definition of the geometrical neighborhood. This technique is equivalent to syntactic structure analysis as developed by Lu and Fu (1978), Sanfeliu et al. (1981), Kayser and Schlegel (1982), and Kayser (1988). Assuming that the texture of a healthy tissue is displayed in the energetically most efficient and stable manner to perform the required biological functions, i.e., to maintain the lowest level of entropy, deviations from this level are reflected in differences in distances and weights between neighboring nuclei or cells. The measure of textural differences in relation to the normal appearance of an organ or tissue is denoted as structural entropy. Since organisms or their compartments are thermodynamically open systems, they are insufficiently described by the (structural) entropy. This parameter only provides a snapshot, with no information about the status of entropy changes from a directed exchange with the environment. The current of entropy, which is equivalent to the amount of entropy exported or imported through a boundary, is an appropriate measure of the "thermodynamic distance" of the system under consideration from its environment, as is readily appreciated for tumors. A solid tumor is a biological system embedded in another one (healthy tissue). Its current of entropy can be calculated if its boundary and proliferative activity are known. This parameter can be measured by histochemical methods (Ki-67 antibody) or from the cytometric characteristics (percentage of S phase-related tumor cells), and the boundary can be measured by the volume fraction of the internal vessels and the size of the external surface of the tumor. Since biochemical factors will contribute to the generation and establishment of these structural and thermodynamic features at the level of tissue organization, histochemical studies can uncover the correlation with these parameter alterations. Taking glycohistochemical determinants as an example for this hypothesis, the potential value of combining the results of immuno /ligandohistochemistry with the data derived from the cytometric or syntactic structure analysis measurements and from the calculations based on thermodynamic theorems is illustrated. PMID- 10370132 TI - Isograft and xenograft of the subcommissural organ into the lateral ventricle of the rat and the formation of Reissner's fiber. AB - The subcommissural organ (SCO) secretes glycoproteins into the cerebrospinal fluid (CSF) that aggregate and form Reissner's fiber (RF). The factors involved in this aggregation are not known. One factor may be the hydrodynamics of the CSF when flowing through the aqueduct. This hypothesis was tested by isografting rat SCO and xenografting bovine SCO into the lateral ventricle of rats. Xenografts were either fresh bovine SCO or explants cultured for 30 days before transplantation. The grafts were investigated by electron microscopy and immunocytochemistry using antibodies against RF glycoproteins, serotonin and the glucose transporter I. Maximal time of transplantation was 43 days for isografts and 14 days for xenografts. The isografts were not reinnervated but were revascularized; they secreted into the ventricle RF glycoproteins that became progressively packed into pre-RF and RF structures identical to those formed by the SCO in situ. RF was confined to the host ventricle and at its distal end the constituent proteins disassembled. Xenografts were neither reinnervated nor revascularized and secreted into the host ventricle a material that never formed an RF. These findings indicate that the CSF factor responsible for the formation of RF is species specific, and that this process does not depend on the hydrodynamics of the CSF. The blood vessels revascularizing the isografted SCO acquired the characteristics of the vessels irrigating the SCO in situ, namely, a tight endothelium displaying glucose transporter I, and a perivascular space containing long-spacing collagen, thus indicating that basal release of glycoproteins may also occur in the grafted SCO. PMID- 10370133 TI - Changes in fibre populations of the rat hairy skin following selective chemodenervation by capsaicin. AB - Perineural application of capsaicin results in a selective and permanent reduction in the sensitivity to noxious chemical and heat stimuli and elimination of the neurogenic inflammatory response. The present quantitative immunohistochemical study has been undertaken to reveal the populations of cutaneous afferent nerves that are affected by perineural capsaicin treatment. Areas of intact and chemodenervated skin were determined with the aid of the vascular labelling technique. In sections taken from intact skin areas, staining with antibodies against protein gene product 9.5 revealed a rich epidermal innervation. Fibres immunoreactive for growth-associated protein 43 were also abundant; nerve fibres immunoreactive for substance P and calcitonin gene-related peptide were less numerous. Somatostatin- and RT97-immunoreactive fibres were seen only in the subepidermal layer. In sections taken from skin areas supplied by the sciatic nerve treated with capsaicin 3 days previously, the number of epidermal nerve fibres immunoreactive to protein gene product 9.5, growth associated protein 43, substance P and calcitonin gene-related peptide was reduced by 90%, 95%, 97% and 66%, respectively. These changes persisted for at least 42 days. The findings reveal that the majority of epidermal axons are capsaicin-sensitive and comprise a chemically heterogeneous population. Reductions in cutaneous fibre populations following perineural capsaicin treatment may result from both the degeneration of sensory axons and the depletion of neuron-specific macromolecules. In addition, most cutaneous nociceptive axons may not use the major sensory neuropeptides substance P and calcitonin gene-related peptide as afferent neurotransmitters. PMID- 10370134 TI - The effects of hibernation on the myenteric plexus of the golden hamster small and large intestine. AB - We have examined the effects of hibernation on the neurochemical composition of myenteric neurones in the small and large intestine of the golden hamster using immunohistochemical and histochemical techniques. Hibernation was induced in golden hamsters by altering the photoperiod and external ambient temperature. Age matched hamsters kept at room temperature and those kept at 5 degrees C but which failed to hibernate were used as controls. Cell counts were carried out to examine possible changes in the numbers of cell bodies immunoreactive to all of the markers examined. The results demonstrated a significant increase during hibernation in the number of neurones immunoreactive to vasoactive intestinal polypeptide, substance P and calcitonin gene-related peptide; cell bodies positive for tyrosine hydroxylase, which were largely absent in the control animals, were prominent in the hibernating animals. There was a significant decrease in the number of neurones immunoreactive to 5-hydroxytryptamine, and no significant changes in the numbers of neurones immunoreactive to protein gene product and nitric oxide synthase. It is suggested that selective upregulation and downregulation of myenteric neurones containing certain neurotransmitters may occur as a protective mechanism during hibernation to maintain the integrity of the muscular and mucosal layers of the intestine in the absence of luminal contents. PMID- 10370135 TI - Differential expression of nitric oxide synthases in EGF-responsive mouse neural precursor cells. AB - Nitric oxide (NO) is a gaseous, radical molecule that plays a role in various physiological processes in the nervous system such as learning and hippocampal plasticity. It is generated from l-arginine by nitric oxide synthases (NOS), which come in three isoforms depending on the tissue of origin, namely inducible NOS (iNOS in macrophages), endothelial-NOS (eNOS in endothelial cells) and neural NOS (nNOS in neural cells). We used epidermal growth factor (EGF)-responsive nestin-positive neural precursor cells originating from the mouse E16 embryonic striatum, and studied the relative expression of NOS isoforms probed with isoform specific antibody using the avidin-biotin immunohistochemical method. Our data revealed both nNOS and eNOS to be expressed in both neurospheres and desegregated neural precursor cells. However, iNOS signals were virtually undetectable in both cell categories. When the neural precursor cells were carried in the presence of poly-l-ornithine (PLO), there was a strong induction of the expression of iNOS proteins, indicating the possibility that this isoform is amenable to modulation by extracellular cues. These preliminary results suggest both nNOS and eNOS to be important in the physiology of neural precursor cells, and that iNOS might also play a role at certain stages in the life of these cells. PMID- 10370136 TI - Tissue expression of the vesicle protein pantophysin. AB - The cell-type restricted expression of cytoplasmic microvesicle membrane protein isoforms may be a consequence of the functional adaptation of these vesicles to the execution of specialized processes in cells of different specialization. To characterize the expression of the vesicle protein pantophysin, an isoform of the synaptic vesicle proteins synaptophysin and synaptoporin, we have prepared and characterized antibodies useful for the immunological detection of pantophysin in vitro and in situ. Using these reagents, we show by immunoblot analyses that pantophysin expression is not homogeneous but differs significantly between various bovine tissues. Furthermore, these differences are not exactly paralleled by the expression of other vesicle proteins of the SCAMP (secretory carrier associated membrane protein) and VAMP (vesicle-associated membrane protein) types that have previously been localized to pantophysin vesicles in cultured cells. By immunofluorescence microscopy, pantophysin expression is seen predominantly in non-neuroendocrine cells with pronounced membrane traffic. Pantophysin staining codistributes with SCAMP and VAMP immunoreactivities in most instances but differs in some. Remarkably, pantophysin staining in liver is restricted to cells surrounding sinusoids and is not detectable in hepatocytes, similar to that of the SCAMP and VAMP isoforms as detected by our reagents in tissue sections. PMID- 10370137 TI - Detection of endogenous biotin in various tissues: novel functions in the hippocampus and implications for its use in avidin-biotin technology. AB - Significant amounts of endogenous biotin were detected by avidin-peroxidase in fixed rat kidney, liver, and brain. The staining was indistinguishable from the true signals of immunoreactivity and could not be consistently blocked by pretreatment with avidin. The finding that certain neurons in the hippocampus contain more biotin than neurons in other areas of the brain suggests that biotin might have novel functions in the brain other than its well-known role as cofactor of carboxylases. Critical examination of published immunohistochemical localization studies on rat kidney strongly suggests that many false-positive results have been considered as true signals. Interference of endogenous biotin in any study using avidin-biotin technology must be considered if biological tissues are involved. The published data obtained by this method should therefore be reevaluated. Furthermore, appropriate controls, blockers and caution in interpreting results must be exercised, not only in immunohistochemistry but also in any applications of avidin-biotin technology. PMID- 10370138 TI - Immunohistochemical evidence for the presence of tryptophan hydroxylase and serotonin in the rodent Harderian gland. AB - The Harderian gland is considered as being an extrapineal source of melatonin. In most rodents, the Harderian gland contains two epithelial cell types (I and II). The aim of this study has been to define which cell type is involved in indoleamine synthesis. The presence and localization of serotonin (melatonin precursor) and tryptophan hydroxylase (the rate-limiting enzyme for serotonin synthesis) have been investigated by immunohistochemistry in male Wistar rats, Syrian hamsters and Djungarian hamsters. The results of the present study show that immunoreactivity for tryptophan hydroxylase and serotonin is confined to the type I cell, suggesting that this cell type is involved in indoleamine synthesis in the rodent Harderian gland. PMID- 10370139 TI - Localization of TFF3 peptide to porcine conjunctival goblet cells. AB - TFF-peptides (formerly P-domain peptides, trefoil factors) form a new family of mucin-associated peptides mainly in the gastrointestinal tract. TFF3 is a typical secretory product of intestinal goblet cells and occurs also in the respiratory tract. Here, polyclonal antisera specific for TFF3 were used in Western blot analysis and immunofluorescence to determine the presence and distribution of TFF3 in the porcine conjunctiva, which is the primary source for ocular mucins. Significant accumulation of TFF3 was detected in conjunctival goblet cells but not in the lacrimal glands. This peptide, together with ocular mucins, may play a role in the rheological function of the tear film. PMID- 10370140 TI - Stretch-induced myogenin, MyoD, and MRF4 expression and acute hypertrophy in quail slow-tonic muscle are not dependent upon satellite cell proliferation. AB - The objectives of these studies were to determine if (1) hypertrophy-stimulated myogenic regulatory factor (MRF) mRNA increases occur in the absence of proliferating satellite cells, and (2) acute hypertrophy occurs without satellite cell proliferation. Adult and aged quails were exposed to 0 or 2500 Rads gamma irradiation, and then wing muscles were stretch-overloaded for 3 or 7 days. MRF mRNA levels in stretch-overloaded and contralateral anterior latissimus dorsi (ALD) muscles were determined after 3 days; hypertrophy was determined after 7 days. The elimination of proliferating cells in irradiated muscles was verified histologically by bromodeoxyuridine incorporation. Relative levels of MRF4, MyoD, and myogenin mRNA were elevated 100%-400% in stretch-overloaded ALD muscles from irradiated adult quails indicating that satellite cell proliferation was not a prerequisite for MRF mRNA increases. Myogenin was the only MRF that exhibited mRNA increases that were lowered by irradiation. This suggests that satellite cells contribute only to myogenin mRNA increases in non-irradiated adult muscles following 3 days of stretch-overload. Stretch-overloaded ALD muscles from aged quails had a relative increase in myogenin mRNA of approximately 150%. The myogenin increase was the same in non-irradiated and irradiated aged animals and also the same as that in stretch-overloaded muscles from irradiated adult quails. Together, these data indicate that attenuated increases in MRF expression in muscles from aged animals are attributable to lower satellite cell MRF expression. ALD muscle masses and protein contents in adult irradiated quails approximately doubled after 7 days of stretch-overload demonstrating hypertrophy despite the elimination of satellite cell proliferation. PMID- 10370141 TI - Sarcomeric myosin heavy chain is degraded by the proteasome. AB - Cardiac myofibrillar proteins, like all other intracellular proteins, are in a dynamic state of continual degradation and resynthesis. The balance between these opposing metabolic processes ultimately determines the number of functional contractile units within each cardiac muscle cell. Although alterations in myofibrillar protein degradation have been shown to contribute to cardiac growth and remodeling, the intracellular proteolytic systems responsible for degrading myofibrillar proteins to their constitutive amino acids are currently unknown. Lactacystin, a recently developed, highly specific proteasome inhibitor, was used in this study to examine the role of the proteasome in myosin heavy chain (MHC) degradation in cultured neonatal rat ventricular myocytes. Cells were treated with growth medium alone or with lactacystin (1-50 microM) for up to 48 h. Lactacystin significantly increased the total protein/DNA ratio and markedly prolonged MHC half-life. Other proteasome inhibitors, namely carbobenzoxy-L leucyl-L-leucyl-L-leucinal (10 microM) and N-acetyl-L-leucyl-L-leucyl-norleucinal (100 microM), were also effective in suppressing MHC degradation. Lactacystin and other proteasome inhibitors also suppressed the markedly accelerated MHC degradation associated with Ca2+ channel blockade but did not prevent the disassembly and loss of myofibrils that accompanied contractile arrest. Thus, sarcomere disassembly precedes the degradation of MHC, which is at least in part mediated by the proteasome. PMID- 10370142 TI - Regulated expression patterns of IRX-2, an Iroquois-class homeobox gene, in the human breast. AB - In the mouse mammary gland, homeobox gene expression patterns suggest roles in development and neoplasia. In the human breast, we now identify a family of Iroquois-class (IRX) homeobox genes. One gene, IRX-2, is expressed in discrete epithelial cell lineages being found in ductal and lobular epithelium, but not in myoepithelium. Expression is absent from associated mesenchymal adipose stroma. During gland development, expression is concentrated in terminal end buds and terminal lobules and is reduced in a subset of epithelial cells during lactation. In contrast to observations for many homeobox genes in the mouse mammary gland in which homeobox gene expression is lost on neoplastic progression, IRX-2 expression is maintained in human mammary neoplasias. Data suggest IRX-2 functions in epithelial cell differentiation and demonstrate regulated expression during ductal and lobular proliferation as well as lactation. PMID- 10370143 TI - Articular chondrocytes and synoviocytes in a co-culture system: influence on reactive oxygen species-induced cytotoxicity and lipid peroxidation. AB - OBJECTIVE: A new co-culture system of rat articular chondrocytes and synoviocytes (HIG-82; cell line) was incubated with phorbol myristate acetate (PMA), H2O2 or a combination of Fe2+ and ascorbic acid to simulate inflammation-like radical attacks in articular joints. METHODS: Chondrocytes were characterized by immunocytochemistry against collagen type II, transmission electron (TEM) and light microscopy. Lipid peroxidation was investigated by measuring thiobarbituric acid-reactive material in the supernatants, cytotoxicity by determining release of lactate dehydrogenase and proliferation by measuring [3H]thymidine incorporation, culture protein and DNA. RESULTS: PMA or Fe2+ and ascorbic acid induced lipid peroxidation in chondrocytes and synoviocytes that was decreased significantly in co-cultures. PMA and H2O2 dose dependently induced release of lactate dehydrogenase in chondrocytes, which was lowered in co-cultures or in previously co-cultured chondrocytes to a nearly basal level. In contrast, conditioned media of synoviocyte cultures showed no lowering effect on the radical-induced toxicity. Protection against H2O2-induced damage of cellular membranes by co-culturing was also shown by TEM. Synoviocytes released chondrocyte-stimulating growth factors spontaneously without previous interaction. CONCLUSION: Chondrocytes establish protective mechanisms against reactive oxygen species via an interaction with synoviocytes. Our co-culture model presents a possible way to study mechanisms of inflammation in articular joints under defined conditions. PMID- 10370144 TI - Beta1-integrins in the cartilage matrix. AB - Integrins are cell-surface receptors that mediate cell attachment to extracellular matrix components. The pericellular matrix in cartilage not only is a mechanical framework, but is also important for chondrocyte differentiation and stabilization of the phenotype. The interaction between chondrocytes and pericellular matrix is mediated, in part, by integrin receptors. We have previously demonstrated the presence of beta1-integrins in the cartilage matrix of organoid culture of limb buds from 12-day-old mouse embryos by immunohistological methods. In order to corroborate these findings, we have further investigated the distribution of integrins in the cartilage matrix by immunoelectron microscopy and by immunoprecipitation methods. Cartilage tissue of limb buds of 17-day-old mouse embryos was treated with collagenase and the cell free and cellular protein-free supernatant was removed and used for immunoprecipitation experiments. Immunoprecipitation with antibodies against beta1-, alpha1-, alpha3-, and alpha5beta1-integrins and collagen type II, followed by immunoblotting with the same antibodies, demonstrated the presence of these integrins and collagen type II in the supernatant. The integrins found in the cartilage matrix could have been either secreted or shed by the cells. The question as to whether they have a function in the cartilage matrix, such as interlinking, in the matrix organization or in the stabilization of matrix components remains to be elucidated. PMID- 10370145 TI - Frozen cultured sheets of human epidermal keratinocytes enhance healing of full thickness wounds in mice. AB - Sheets of cultured allogeneic human keratinocytes have been used for the treatment of burns and chronic leg ulcers but there has been no animal assay for the therapeutic action of these cultures. In order to analyze the effects of frozen cultures of human keratinocytes on wound healing, we have developed such an assay based on the rate of repair of full-thickness skin wounds in immunocompetent NMR1 mice. Reepithelialization of the control wounds, originating from the murine epithelium at the edge of the wound, occurred at a constant rate of advance of 150 microm/day. When frozen cultured human epidermal sheets were thawed at room temperature for 5-10 min and applied to the surface of the wound, the murine epithelium advanced at 267 microm/day. Most wounds treated with frozen cultures completely healed after 10 days, whereas most control wounds required 16 days. The accelerated reepithelialization did not depend on the presence of proliferative human keratinocytes in the frozen cultures. The cultures also promoted early formation of granulation tissue and laminin deposition over the surface of the wound bed. This simple assay should permit quantitative analysis of the effects on healing exerted not only by cultured cells, but also by proteins and small molecules. PMID- 10370146 TI - Immunolocalization of hyaluronic acid and inter-alpha-trypsin inhibitor in mice. AB - The direct interaction of hyaluronic acid (HA) and heavy chain (HC) of the inter alpha-trypsin inhibitor (IalphaI) family plays a critical role in the organization and stabilization of the extracellular matrix. The aim of the present investigation was to elucidate the distribution of the IalphaI HC and HA in adult mouse tissues. An immunohistochemical method using a rabbit polyclonal antibody raised against mouse IalphaI heavy-chain peptide and a specific probe for HA (biotinylated HA-binding protein) was used to demonstrate an immunolocalization of IalphaI HC and HA. Distribution and localization of HA was of three types, namely, colocalization with IalphaI HC itself (cartilaginous tissue and ovary), localization around IalphaI HC immunostaining (lung, intestine and skeletal muscle), and localization at a small distance from IalphaI HC or a different distribution pattern (brain, liver, skin and kidney). These results indicate that IalphaI HC could function as an HA-rich matrix stabilizer on the cells of cartilage and maturing ovary, in which IalphaI HC shows colocalization with its predominant ligand, HA. PMID- 10370147 TI - Localisation of P2X5 and P2X7 receptors by immunohistochemistry in rat stratified squamous epithelia. AB - In order to investigate whether purinoceptors are involved in the physiological renewal and regeneration of epithelia, we used immunohistochemical techniques on fresh frozen sections of various stratified squamous epithelial tissues (cornea, tongue, soft palate, oesophagus, vagina and footpad) of the rat and specific polyclonal antibodies to unique peptide sequences of P2X1-7 receptors. Only two of the antibodies, anti-P2X5 and anti-P2X7, reacted with epithelial structures. P2X5 immunoreactivity was mainly associated with the membranes of the proliferating and differentiating cell layers (spinous and granular layer) in both keratinised and non-keratinised epithelia and growing hair follicles. In contrast, P2X7 immunoreactivity was clearly associated with the keratinisation process, the staining being most intense in the upper keratinised and the exfoliated layers. These findings suggest, for the first time, that P2X5 and P2X7 receptors play an important role in the physiological turnover of continuously regenerating cells, and further, raise the possibility that they represent novel targets for the development of pharmacological tools of potential benefit for diseases of epithelial dysfunction. PMID- 10370148 TI - The antisecretory factor: synthesis and intracellular localisation in porcine tissues. AB - The antisecretory factor, AF, is a 41-kDa protein, cloned and sequenced from a human pituitary library. AF is a potent inhibitor of experimental intestinal hypersecretion in rats and pigs. An antiserum against the C-terminal of the truncated, recombinantly produced AF protein was raised in rabbits. The affinity purified antiserum was used to study the expression of AF in mucosal membranes and in the pituitary gland of the pig; distinctly stained cells were found in lymphoid cells in the connective tissue of all parts of the gastrointestinal, respiratory and urinary tracts. Cytoplasmic AF was demonstrated in endocrine and epithelial cells in the pituitary gland. In situ hybridisation with a digoxigenin labelled mRNA probe also demonstrated specific cytoplasmic staining in epithelial and lymphoid cells in all of these tissues. The cells stained by either method were similarly distributed topographically within the tissues. The results suggest that a specific defined cell population in these various tissues possesses the capability of both synthesising and storing the AF protein within the cellular cytoplasmic compartment. PMID- 10370149 TI - Development of the wing discs of Zophobas atratus under natural and experimental conditions: occurrence of a gradual larval-pupal commitment in the epidermis of tenebrionid beetles. AB - Using light and electron microscopy, we studied the development of the wing discs in the large beetle Zophobas atratus, under natural and experimental conditions. A reversible differentiation of the wing discs is usually observed during supernumerary instars of crowded larvae. Juvenile hormone analog (JHA) application during the wandering period or compelled experimental crowding during the larval-pupal switchover - or commitment - inhibits the onset of metamorphosis. Isolation, followed by recrowding, also induces the disc cells to secrete unusual cuticular material. Recrowding is able to trigger the reversal of metamorphosis during the 4-day period when larval-pupal commitment is taking place. Likewise, feeding behaviour which normally stops at commitment often recovers. Ecdysis of intermediate instar animals (prothetelic larvae) corroborates the occurrence of a temporal and spatial variation to commitment, unique to each organ. All these data lead us to consider this 4-day period, which we have called the C period or commitment period, extending from the wandering stage (the previous T period) to the crooked posture stage (i.e. from eyestage 4 to 7) as the physiological time during which the larval organs are gradually committed to differentiate into pupal organs. PMID- 10370150 TI - Presumptive insect circadian pacemakers in vitro: immunocytochemical characterization of cultured pigment-dispersing hormone-immunoreactive neurons of Leucophaea maderae. AB - The accessory medulla with its associated pigment-dispersing hormone immunoreactive neurons appears to be the pacemaker that controls the circadian locomotor activity rhythm of the cockroach Leucophaea maderae. To permit studies at the level of individual, identified, pacemaker neurons, we developed specific long-term primary cell cultures of fully differentiated adult neurons of the accessory medulla. As judged from soma diameter distribution, the cultures contain an unbiased representation of apparently all neuronal types of the accessory medulla. The cultured cells survive and grow processes for more than 2 months with or without additional hemocyte coculturing. However, a strong positive effect on initial outgrowth was observed with hemocyte coculturing. At least six different morphological cell types of the accessory medulla could be distinguished in vitro. Among these only one cell type, the monopolar type C cell, was recognized in vitro with an antiserum against the neuropeptide pigment dispersing hormone. Thus, the identifiable monopolar type C cells are candidates for circadian pacemaker neurons and will be the focus of further physiological characterizations. PMID- 10370151 TI - Types, numbers and distribution of synapses on the dendritic tree of an identified visual interneuron in the brain of the locust. AB - The descending contralateral movement detector (DCMD), an identified descending interneuron in the brain of the locust Schistocerca gregaria has been investigated by using light and electron microscopy. We describe the fine structure, distribution and numbers of synapses that it receives from another identified brain neuron, the lobular giant movement detector (LGMD), and from unidentified neurons. The DCMD dendrites emerging from the integrative segment vary in form and number between individuals and sexes but always form a flattened dendritic domain. The arborizations and the integrative segment appear to be exclusively postsynaptic. Two types of synaptic contacts (Type 1 and 2) onto the DCMD can be discerned as having either round (Type 1) or pleiomorphic synaptic vesicles (Type 2) and by large (Type 1) or small (Type 2) subsynaptic appositions. Contact zones of Type 1 synapses are smaller than those of Type 2. LGMD-synapses are of Type 1 and occur intermingled with presynaptic sites of unidentified units. Some branches of the DCMD receiving input from unidentified units are devoid of contacting LGMD processes. Synapses of both types are randomly distributed over the DCMD integrative segment and at fibres with similar sizes. Type 1 synapses are much more frequent than Type 2 synapses and their number is negatively correlated with fibre diameter. For a whole DCMD dendritic arborization, a total of 8500 active zones of chemical synapses has been calculated, including a minimum of 2250 LGMD-synapses and about 1000 Type 2 synapses. The DCMD may thus receive a considerable amount of input from as yet unidentified neurons. PMID- 10370152 TI - Differentiation of pinopsin-immunoreactive cells in the developing quail pineal organ: an in-vivo and in-vitro immunohistochemical study. AB - The avian pineal organ contains several types of photoreceptors with different photopigments: rhodopsin, iodopsin, and pinopsin. We have previously examined the differentiation of both rhodopsin-like and iodopsin-like immunoreactive cells during pineal development in quail embryos to determine the onset of synthesis of specific proteins and their cellular localization. In the present study, we have performed pinopsin immunohistochemistry on in-vivo developing and in-vitro cultured pineal organs of quail embryos. The results were compared with those obtained with rhodopsin and iodopsin immunohistochemistry. In the developing pineal organs, pinopsin immunoreactivity was detected at embryonic day 8, i.e. five days earlier than rhodopsin-like and iodopsin-like immunoreactivities. It was localized exclusively in the protrusions extending into the lumen throughout development, whereas rhodopsin-like and iodopsin-like immunoreactivities were usually found both in cell bodies and processes. These differences were also observed under two different types of culture conditions (dissociated cell culture and organ culture) indicating that, in the avian pineal organ, the expression pattern of the pinopsin gene is basically different from those of the other two pineal photopigments. The present study suggests that pineal cells have a mechanism for the polarized transport of pinopsin molecules. PMID- 10370154 TI - Orthopedic breakthroughs dispatched from Yokohama: from 19th-century developments to 21st-century prospects. PMID- 10370153 TI - Elongated pericyte-like cells connect discrete capillaries in the cochlear stria vascularis of gerbils and rats. AB - Bridging structures between discrete capillaries in the stria vascularis of the cochlea were studied morphologically in gerbils and rats. Serial thin sections for transmission electron microscopy revealed (1) that elongated cells surrounded by the basal lamina provided the structural basis for the bridging structure, (2) that the basal lamina surrounding the elongated cell extended to the basal lamina around the capillary endothelial cell, (3) that the electron density of the cytoplasm was similar to that of the pericytes around the capillaries, and (4) that the cell was attached to the capillaries at both ends only. Visualization of the basal lamina by immunofluorescent methods revealed (1) that capillaries were often bent at the site of attachment of the bridging cell, (2) that the bridging cell bifurcated occasionally, and (3) that the density of the bridging cell was much higher in the stria vascularis than in the underlying spiral ligament. Filamentous actin visualized by fluorescent phalloidin was not apparent in the bridging cell. We propose that the bridging cell provides mechanical strength to the tortuous capillary network in the stria vascularis and participates in the specific function of the stria vascularis in cooperation with other types of cells. PMID- 10370155 TI - Remodeling of patellar tendon grafts augmented with woven polyester after anterior cruciate ligament reconstruction in humans. AB - Arthroscopic and histological evaluations of anterior cruciate ligament (ACL) grafts were made in 38 patients at various intervals after reconstruction was performed with patellar tendons (PTs) augmented with woven polyester. The interval between the surgery and the examinations ranged from 3 to 36 months (mean, 14.1 months). Biopsy specimens with woven polyester were removed from the anterior mid-portion of the graft 5 mm in depth from the graft surface. Arthroscopically, all 38 grafts showed thick ligamentous tissues, but some polyester fibers had ruptured in 9 grafts. The transplanted grafts were arthroscopically very similar to normal ACL 12 months after reconstruction. Histologically, a marked inflammatory response to the woven polyester was observed in the first 4 months after the operation, but this decreased with time. Five of 6 grafts obtained 18-36 months postoperatively showed decreased cellularity, with longitudinally oriented fibrous bundles, suggesting approaching ligament maturity. The appearance of the fibroblasts in the grafts varied from patient to patient, with 13 of 37 grafts (35.1%) showing spindling fibroblasts, and 24 grafts (64.9%) oval ones. (The remaining graft consisted of mesenchynal cells). Clinically, postoperative KT 1000 arthrometer measurements showed that the mean injured-to-uninjured difference in anterior knee laxity was 1.7 +/- 2.6 mm. There was no chronic synovitis or effusion during the study period. These results suggest that grafts with PT augmented with woven polyester transform arthroscopically and histologically into a structure similar to that of normal ligaments after reconstruction, and may act as a substitute for the ACL. The advantage of woven polyester for the augmentation of PT grafts was that it allowed earlier mobilization and weight-bearing after reconstruction, without adverse effects on graft integrity. PMID- 10370156 TI - Analysis of synovial fluid components of hydrarthrosis in long-term hemodialysis patients. AB - The synovial fluid components in long-term hemodialysis patients (HD; 43 knees in 43 patients) were investigated and compared with those in patients with osteoarthritis (OA; 21 knees in 21 patients) and rheumatoid arthritis (RA; 26 knees in 26 patients). The average ages in the three groups were, respectively, 60.7 years (range, 34-79 years), 63.2 years (range, 48-88 years), and 59.7 years (range, 37-76 years). The duration of hemodialysis in the HD group averaged 14.0 years (range, 4-24 years). The concentrations of hyaluronic acid, protein, and isomers of chondroitin sulfate (chondroitin 6-sulfate [C6S] and chondroitin 4 sulfate [C4S]) in the synovial fluid, and its viscosity were measured. Differences in each of the parameters were investigated according to disease clinical stage, roentgenological grade, and periods of dialysis in the HD group. The viscosity of the synovial fluid and the concentration of hyaluronic acid in HD patients were similar to those in OA patients; however, the C6S/C4S ratio in the synovial fluid of HD patients was similar to that in RA patients. The latter finding suggests that synovitis may be present in the hydrarthrosis of HD patients. The cause of this synovitis in HD patients remains to be elucidated. PMID- 10370157 TI - New method of assessing outcome of treatment for cervical myelopathy (M/T method) -preliminary report. AB - In some patients with cervical radiculomyelopathy, neurological examination reveals extreme muscle weakness in an upper extremity with no, or a relatively insignificant, sensory deficit. In assessment of treatment outcome in these patients, the previous method used is thought to be unsuitable. We developed a new method of assessment (the M/T method) in which the recovery of upper extremity muscle weakness is assessed. The purpose of this study was to evaluate this new method. In the M/T method, change in the function of the spinal cord, including the anterior root, is expressed as an index number (recovery rate) in relation to the time-course. The function of the nerve tissue is expressed as the value for the manual muscle testing (MMT) of muscle power, and the muscle tested is represented by one that has the least MMT value before treatment. The unit of measurement of the time-course is 3 months. The index is the difference between the MMT value at the time of follow-up and that before treatment, divided by the number of time units after treatment. The index can be expressed in both fractional and decimal forms. Fractions are useful for examining an individual patient's progress and the decimal form is useful for comparing the data of multiple patients. The M/T method was employed in 16 patients (13 men, 3 women; age, 45-79 years; follow-up, 6 months - 8 years) whose chief complaints were muscle weakness in the upper extremities and who had undergone double-door laminoplasty. Seven patients had an M/T index of 1 or more; the score was 4 in one patient, 2 in four patients, and 1 in two patients. Two patients had an M/T index between 0 and 1; 0.17 and 0.25. Five patients had an index value of 0. The M/T index in two patients was less than 0; -0. 17 and -0.11. Using the M/T method, recovery can be presented as an index number, and as a result, it is easy to compare differences in the recovery rate among patients. The M/T method is useful for evaluation of the viability of the spinal cord, including the anterior root; in particular for those patients in whom neurological examination reveals extreme muscle weakness in an upper extremity with no, or a relatively insignificant, sensory deficit. PMID- 10370158 TI - Is bone density in the distal femur affected by use of cement and by femoral component design in total knee arthroplasty? AB - The objective of this study was to evaluate changes in bone density in the distal femur 2 years after total knee arthroplasty with four different implant designs using cemented or cementless femoral components. Bone density was measured retrospectively from radiographs of 114 knees, using a densitometer. A decrease in bone density of up to 57% was identified in the distal femora with a cemented femoral component 2 years after surgery, compared with a decrease of up to 28% with a cementless, porous-coated component of the same design. The differences between the four implant designs in the changes in bone density in the anterior distal femur were significant at 2 years (P < 0.001). A possible cause of this result may be a difference in load transfer due to the different methods of fixation. The surgeon should expect decreased bone density in the distal femur at the time of revision surgery, especially with a cemented femoral component. PMID- 10370159 TI - Electromyographic evaluation after endoscopic carpal tunnel release in idiopathic carpal tunnel syndrome. AB - The purpose of this study was to electromyographically evaluate results in patients with carpal tunnel syndrome (CTS) who underwent endoscopic carpal tunnel release (ECTR). The subjects were 26 patients with idiopathic CTS (37 hands) who were followed for at least 6 months after ECTR. To compare results informatively, hands were classified into four groups: those with normal distal motor latency (DML) and sensory conduction velocity (SCV) were classified as group A, those with normal DML and abnormal SCV as group B, those with an abnormal DML and normal SCV as group C, and those with abnormal DML and SCV as group D. All but one of the hands were classified as group D on the basis of preoperative electromyographic evaluation, while one was classified as group C. The mean preoperative obtainable DML and SCV values were 7.2 m and 27.3 m/s, respectively. Postoperatively, 12 hands were in group A, 8 hands in group B, 2 hands in group C, and 15 hands in group D. The mean DML and SCV values at final follow-up were 4.3 ms and 40.8 m/s, respectively. Of the 25 hands with muscle atrophy before surgery, 6 hands were in group A, 5 hands were in group B, 1 hand was in group C, and 13 hands were in group D at final follow-up. Thenar muscle atrophy and denervation potentials were present before surgery in 13 of the 15 hands classified as group D at the final follow-up. PMID- 10370160 TI - Anterior cruciate ligament reconstruction and physiological joint laxity: earliest changes in joint stability and stiffness after reconstruction. AB - Physiological joint laxity is an important element of normal knee joint function, providing smooth joint movement. However, the objective evaluation of post operative results after knee ligament surgery is usually based primarily on stability and range of motion, and joint laxity has been ignored. In this study, we measured the joint stiffness of 82 knees undergoing anterior cruciate ligament (ACL) reconstruction with the Leeds-Keio artificial ligament, before the operation, immediately after the operation, and finally when the full range of motion was achieved postoperatively; changes in joint laxity after the ACL reconstruction were investigated. Before the operation, joint laxity was greater than that of the normal side (P < 0.01), but immediately after the operation it diminished compared not only with that observed preoperatively, but also with that of the normal side. When the full range of motion was achieved, joint laxity was lower than that observed immediately after the operation (P < 0.01), but still remained higher than that of the normal side (P < 0.01). In other words, stability was achieved, but joint laxity was diminished through the operation. In this series, a stiffer artificial ligament than the natural ACL was used, and maximum tension was applied during the operation, aiming at better stability, but this may cause diminution of joint laxity. PMID- 10370161 TI - Accurate pedicle screw insertion under the control of a computer-assisted image guiding system: laboratory test and clinical study. AB - We used a commercially available computer-assisted navigation system (StealthStation; Sofamor Danek, Memphis, TN, USA) in both an in-vitro and a clinical study performed in 1996-1998. The basic data used for navigation were preoperative computed tomography (CT) scan imaging data. The position of the probe or drill guide was superimposed in real-time on a monitor. For the in-vitro study, ten plastic lumbar spine models (50 vertebrae) were used. The entrance hole for the screw was made by drilling, following navigation. Using the navigation system, we drilled 88 holes through the pedicles into the vertebral bodies of 44 vertebral models. All 88 pedicle holes were contained within the pedicle without perforation. The mean deviation of the hole positions from the surgical plan was 1.78 +/- 0.81 mm, and the mean angular deviation was 2.28 degrees +/- 1.92 degrees. In 29 patients, using the navigation system, we introduced 169 pedicle screws at the planned position. Fifty-one screws were used for thoracic and 118 screws for lumbar spinal fixation. All screws correctly passed through the pedicles. There were no neurological complications after surgery. Using this guided surgery system, we achieved satisfactory results both in the laboratory and in a clinical setting. PMID- 10370162 TI - An operative procedure for advanced Kienbock's disease. Excision of the lunate and subsequent replacement with a tendon-ball implant. AB - Excision of the lunate and subsequent replacement with a tendon implant was performed in 22 patients with Kienbock's disease between 1971 and 1985. This procedure was indicated mainly for those with advanced Kienbock's disease, i.e., stage III or IV according to the Lichtman classification. After the collapsed lunate is removed, a tendon-ball implant, made of the palmaris longus and plantaris tendons is placed in the resultant space in the carpus. A forearm distractor is applied during the operation, and distraction is continued for 4 weeks postoperatively. We report the long-term results in 15 patients, whose average follow-up period was 16 years and 3 months. One patient with infection was excluded from the study because the implanted tendon was removed 2 weeks after the operation, and 6 patients were lost to follow-up. All patients were free of pain after the surgery. The flexion-extension range of the wrist increased by 14.2 degrees, on average, after the surgery. The average grip power of the operated hand was 90.2% of that in the non-operated hand. Calcification and ossification were frequent in the implanted tendons a few months postoperatively. The average carpal height ratio (defined as carpal height/length of the third metacarpal) was 0.53 before the operation and 0.49 at the time of follow-up. According to Dornan's classification of clinical results, 9 of the 15 patients were classified as having excellent results and 6 as good. PMID- 10370163 TI - Measurement of bone marrow blood volume in the knee by positron emission tomography. AB - Positron emission tomography was used to measure bone marrow blood volume (BBV), an important hemodynamics parameter, in the knee. The subjects were 11 healthy male volunteers (mean age, 23.6 years; range, 21-27 years). The 15O-labelled carbon monooxide (C15O) single-breath inhalation method was used. In the distal femur, regional (r) BBV in the posterior area of the epiphysis (medial, 2. 25 ml/100 cm3 bone marrow; lateral, 2.03 ml/100 cm3) was significantly less than that in the anterior area of the epiphysis (medial, 3.48 ml/100 cm3; lateral, 3.01 ml/100 cm3) and that in the metaphysis-to-distal diaphysis (2.90-3.67 ml/100 cm3). In the proximal tibia, rBBVs in the metaphysis-to-proximal diaphysis (2. 32 2.76 ml/100 cm3) were significantly less than those in the area of the physis (medial, 3.30 ml/100 cm3; lateral, 3.53 ml/100 cm3). These regional differences in rBBV within the knee may be associated with the development of ischemic bone marrow disorders, such as steroid-induced osteonecrosis, in the knee. PMID- 10370164 TI - Microsatellite alterations in various sarcomas in Japanese patients. AB - To determine the usefulness of microsatellite analysis in the differential diagnosis of various sarcomas, we investigated microsatellite alterations at 12 microsatellite loci by polymerase chain reaction and electrophoresis in 39 Japanese patients with sarcomas. The sarcomas were: osteosarcoma, Ewing's sarcoma, chondrosarcoma, liposarcoma, leiomyosarcoma, epithelioid leiomyosarcoma, rhabdomyosarcoma, synovial sarcoma, and malignant fibrous histiocytoma. We also examined ten leiomyomas to contrast with leiomyosarcoma. No microsatellite instability (MSI) or loss of heterozygosity (LOH) were found in Ewing's sarcoma, chondrosarcoma, epithelioid leiomyosarcoma, malignant fibrous histiocytoma, and leiomyoma. Only three patients, one each with liposarcoma, leiomyosarcoma, and synovial sarcoma, manifested MSI, whereas, osteosarcoma, liposarcoma, leiomyosarcoma, rhabdomyosarcoma, and synovial sarcoma manifested LOH, with an incidence of 43%, 14%, 86%, 20%, and 75%, respectively. Interestingly, three patients showed unusual patterns of LOH, probably due to intratumoral heterogeneity. Kaplan-Meier analysis revealed that LOH on 11p was predictive of poor prognosis in osteosarcoma. The low incidence of MSI indicates that MSI is not necessary for neoplastic transformation in sarcomas. However, the very high incidence of LOH in leiomyosarcoma indicates that microsatellite analysis may serve for the differential diagnosis of leiomyosarcoma versus leiomyoma. Microsatellite analysis may also predict prognosis in osteosarcoma. PMID- 10370165 TI - Effects of 4-hydroperoxy ifosfamide in combination with other anticancer agents on human cancer cell lines. AB - Ifosfamide is one of the currently available anticancer agents with a broad spectrum of clinical activity against a variety of tumors. To investigate its optimal combinations, we studied the effect of 4-hydroperoxy ifosfamide (the active form of ifosfamide) in combination with other anticancer agents against two human cancer cell lines, MG-63 (an osteosarcoma cell line) and MOLT-3 cells (a T-cell leukemia cell line). The cells were incubated for 4 days and 3 days, respectively, in the presence of 4-hydroperoxy ifosfamide and the other agent. Cell growth inhibition was determined by MTT assay. The effects of these drug combinations at the concentration producing 50% inhibition (IC50) were analyzed by the isobologram method. 4-Hydroperoxy ifosfamide showed additive effects with bleomycin, cisplatin, cytarabine, doxorubicin, etoposide, 5-fluorouracil, and mitomycin C, while it showed a protective effect with methotrexate in both cell lines. 4-Hydroperoxy ifosfamide showed an additive effect with vincristine in the MG-63 cell line, while it showed a sub-additive effect in the MOLT-3 cell line. No anticancer agents tested showed a supra-additive effect with 4-hydroperoxy ifosfamide. These data suggest that ifosfamide is advantageous for simultaneous administration with a majority of the anticancer agents we studied. Methotrexate is an inappropriate drug for simultaneous administration with ifosfamide. PMID- 10370166 TI - Transdermal iontophoretic delivery of enoxacin from various liposome-encapsulated formulations. AB - The major purpose of this work was to study the effect of various liposome formulations on the iontophoretic transport of enoxacin through excised rat skin. The electrochemical stability of these liposomes was also evaluated. The encapsulation percentage of enoxacin was significantly enhanced after 6 h incubation in an electric field; whereas the fusion of liposomes was inhibited by application of electric current. The results of iontophoretic drug transport showed that the permeability of enoxacin released from liposomes was higher compared with that of free drug. The iontophoretic permeability of enoxacin released from liposomes increased with a decrease in the fatty acid chain length of the phospholipid, which may be due to the different phase transition temperatures of the phospholipids. Incorporation of charged phospholipid resulted in an alteration of the transdermal behavior of enoxacin: the iontophoretic permeation as well as the amount of enoxacin partitioned in skin was greatly reduced after incorporation of stearylamine in liposomes, which can be attributed to the competitive ion effect. The enoxacin released from stratum corneum-based liposomes showed the highest amount of enoxacin partitioned into skin depot. The results of employing cathodal iontophoresis on negative charged liposomes suggested that the liposomal vesicles or phospholipids may carry enoxacin into deeper skin strata via the follicular route. PMID- 10370167 TI - Poly(fumaric-co-sebacic anhydride). A degradation study as evaluated by FTIR, DSC, GPC and X-ray diffraction. AB - The degradation of three poly(fumaric-co-sebacic anhydride) [P(FA:SA)] copolymers is examined in a composition of microspheres made by the hot melt encapsulation process. The emergence of low molecular weight oligomers occurs during degradation of the copolymer microspheres, as evidenced by a variety of characterization methods. Characterization was conducted to determine the extent of degradation of the polyanhydride microspheres using Fourier-transform infrared spectroscopy (FTIR), gel permeation chromatography (GPC), differential scanning calorimetry (DSC) and X-ray diffraction. It is demonstrated that degradation of P(FA:SA) is greatly accelerated at basic pH, yet there is little difference between degradation in neutral and acidic buffers. A good correlation exists between the results of each characterization method, which allows a better understanding of the degradation process and the resulting formation of low molecular weight oligomers in poly(fumaric-co-sebacic anhydride). PMID- 10370168 TI - Factors influencing nonoxynol-9 permeation and bioactivity in cervical mucus. AB - The effects of delivery gel pH and osmolarity on both the mass transport and 'biodiffusion' of the spermicide nonoxynol-9 (N9) in bovine cervical mucus were evaluated. Delivery gels were calcium chloride crosslinked alginate containing 3% N9, and were manufactured over a pH range of 3.4 to 5.9 and an osmolarity range of 300 to 900 mosmol. Mass transfer parameters (diffusion coefficients and total drug loading) were determined using a new UV spectrophotometric technique while biodiffusion (the diffusion distance into mucus at which sperm are killed) was assessed using the Double Ended Test. It was found that delivery gel pH had a significant effect on spermicidal efficacy of the alginate-N9 system; biodiffusion increased with decreasing pH. Actual N9 diffusion into mucus was found to be influenced by both the delivery gel pH and osmolarity. At high N9 concentration (near the gel/mucus interface), mass transport tended to decrease with decreasing pH at the highest osmolarity. At low concentration, mass transport tended to decrease with increasing osmolarity and decrease with increasing pH at the highest osmolarity. The difference between low and high concentration behavior can be attributed to N9 micelle formation. These findings are interpreted in the context of the design of intravaginal drug delivery vehicles for spermicides. PMID- 10370169 TI - Comparison of the effects of short, high-voltage and long, medium-voltage pulses on skin electrical and transport properties. AB - High-voltage pulses have been shown to increase rates of transport across skin by several orders of magnitude on a time scale of minutes to seconds. Two main pulse protocols have been employed to promote transport: the intermittent application of short ( approximately 1 ms) high-voltage (approximately 100 V across skin) pulses and a few applications of long (=100 ms) medium-voltage (>30 V across skin) pulses. In order to better evaluate the benefits of each protocol for transdermal drug delivery, we compared these two protocols in vitro in terms of changes in skin electrical properties and transport of sulforhodamine, a fluorescent polar molecule of 607 g/mol and a charge of -1. Whereas both protocols induced similar alterations and recovery processes of skin electrical resistance, long pulses of medium-voltage appeared to be more efficient in transporting molecules across skin. Skin resistance decreased by three (short pulses) and two (long pulses) orders of magnitude, followed by incomplete recovery in both cases. For the same total transported charge, long pulses induced faster and greater molecular transport across skin than short pulses. In addition, a greater fraction of the aqueous pathways created by the electric field was involved in molecular transport when using long pulse protocols. Transport was concentrated in localized transport regions (LTRs) for both protocols but LTRs created by long pulses were an order of magnitude larger than those formed by short pulses and the short pulses created an order of magnitude more LTRs. Overall, this study is consistent with the creation of fewer, but larger aqueous pathways by long, medium-voltage pulses in comparison to short, high-voltage pulses. PMID- 10370170 TI - Evaluation of the use of xanthan as vehicle for cationic antifungal peptides. AB - Oral candidiasis frequently occurs in individuals with dry mouth syndrome (xerostomia), in immunocompromised patients and in denture wearers. The aim of this study was to develop a formulation which will prolong the retention time of antimicrobial agents at the site of application. The activity against Candida albicans of a synthetic cationic peptide dhvar 1, based on the human fungicidal salivary peptide histatin 5, was tested either in a mixture with the bioadhesive polymer xanthan, or after covalent coupling to this polymer. The presence of xanthan resulted in an increase of the LC50 value of the peptide from 2.6 (S.D.=0.6) to 5.8 (S.D.=4.0). Covalent coupling caused an additional increase of the LC50 value to 18.4 (S. D.=6.7). Coupling caused a reduction of the viscosity and elasticity of the xanthan solution related to the applied concentration of the coupling agent. Incubation of the peptide with clarified human whole saliva resulted in proteolytic degradation of the peptide. In the presence of xanthan the degradation occurred more slowly. It was concluded that xanthan is an appropriate vehicle for antimicrobial peptides in a retention increasing formulation. PMID- 10370171 TI - Sol-gel composite films with controlled release of biocides. AB - The release of biocides (benzoic, sorbic and boric acids) incorporated into modified silica films was investigated with respect to composite structure. The liberation rates of the embedded acids are proportional to the biocide-to-silica ratio and are changed by adding soluble polymers such as hydroxypropylcellulose. The rates of liberation correlate with biocidal activity, i.e., the growth of microorganisms such as Escherichia coli, Lactobacillus plantarum and Penicillium sp. is strongly suppressed by contact with such composite films. In a similar way, strong fungicide and insecticide effects were observed after impregnating wood with composites containing boric acid. PMID- 10370172 TI - Percutaneous absorption of sunscreens from liquid crystalline phases. AB - The purpose of the present study was to investigate the effects of two nonionic surfactants with liquid crystalline structures on the cutaneous availability of two sunscreens. Three liquid crystalline structures were investigated: lamellar, hexagonal and cubic. The diffusion of sunscreens within the liquid crystals was determined by measuring transport kinetics into an unloaded surfactant medium from a similar system loaded with the sunscreens. The diffusion coefficients were the greatest in the cubic systems for benzophenone-4 (a hydrosoluble sunscreen) and in lamellar systems for octyl methoxycinnamate (a liposoluble sunscreen). So the diffusion in this surfactant systems was strongly dependent on the structure of the liquid crystal and on the physicochemical properties of the solute. The transcutaneous fluxes were determined using a Franz-type diffusion cell. The liquid crystalline vehicles modified the transcutaneous fluxes of benzophenone-4 but did not change those of octyl methoxycinnamate. The solute diffusion within the vehicle was not the rate-determining step for transcutaneous permeation for either sunscreen. The diffusion of benzopenone-4 within the stratum corneum and that of octyl methoxycinnamate within the dermis could be the rate-determining steps for their transcutaneous permeation. These two steps could be affected differently by nonionic surfactant vehicles. PMID- 10370173 TI - Thermodynamic prediction of active ingredient loading in polymeric microparticles. AB - The growing use of microparticles as a controlled-delivery system for pharmaceutical and non-pharmaceutical active ingredients (AIs) has prompted a costly trial-and-error development of new and effective microparticle systems. In order to facilitate a more rational design and optimization of AI loadings in microparticles, we have developed a molecular-thermodynamic theory to predict the loading of liquid AIs in polymeric microparticles that are manufactured by a solvent evaporation process. This process involves the emulsification of a liquid polymer solution (consisting of polymer and AI dissolved in a volatile solvent) in an aqueous surfactant solution. The theory describes the equilibrium distribution of the AI between the aqueous phase and the dispersed polymeric droplets. The universal functional activity coefficient (UNIFAC) and UNIFAC-Free Volume (FV) group-contribution methods are utilized to model the nonidealities in the water and polymeric droplet phases, respectively. The inputs to the theory are: (i) the chemical structures, densities and total masses of the manufacturing ingredients, (ii) the manufacturing temperature and (iii) the glass transition temperature of the polymer. Since surfactant concentrations exceeding the critical micellar concentration (CMC) are often required in order to stabilize the dispersed polymeric droplets during the emulsion manufacturing process, the theory also accounts for AI solubilization in surfactant micelles present in the manufacturing solution. To test the AI loading predictions, we compare theoretical predictions of AI loadings in poly(lactic acid), poly(methyl methacrylate) and polystyrene microparticles to experimentally measured ones for five model AIs with varying degrees of hydrophobicity (benzyl alcohol, n-octanol, geraniol, farnesol and galaxolide). We also demonstrate how the developed theory can be utilized to screen polymers with respect to their abilities to load a given AI, as well as to provide guidelines for manufacturing microparticles having the desired AI loading. PMID- 10370174 TI - Spatially constrained localized transport regions due to skin electroporation. AB - Rapid, controlled molecular transport across human skin is of great interest for transdermal drug delivery and minimally invasive chemical sensing. Short, high voltage pulses have been shown previously to create localized transport regions in the skin. Here, we show that these regions can be constrained to occur at specific sites using electrically insulating masks that restrict the field lines. The increase in total ionic and molecular transport per area was comparable to the levels observed in unconstrained electroporation of human skin. Constraining the area of intervention to encompass small areas of interest, a primary feature in the design of microdevices for transdermal drug delivery, can provide the same levels of flux as the unconstrained case. PMID- 10370175 TI - A dose-response study of epidural liposomal bupivacaine in rabbits. AB - Liposomes are drug delivery systems used to prolong local effects of bupivacaine. We studied the relationships between motor and hemodynamic changes and epidural doses of plain bupivacaine (P) and liposomal bupivacaine (L) in rabbits equipped with chronical lumbar epidural and femoral arterial catheters. Liposomal (phosphatidylcholine-cholesterol) suspensions contained 20 mg ml-1 of lipid, and different doses of bupivacaine (Lipo 7.5=7.5-; Lipo 3.7=3. 75-; Lipo 2.5=2.5-; Lipo 1.2=1.25-; and Lipo 0.7=0.65-mg of bupivacaine per ml). Forty rabbits were randomly assigned to five groups to receive epidural anesthesia (1 ml) as follows: Groups I to V received 0.65 to 7.5 mg of bupivacaine as P then as L. Release rate of bupivacaine from liposome was significantly slower using Lipo 3.7 than after Lipo 2.5 (Td was 3.9 h and 1.7 h respectively). Increasing the doses of L and P resulted in faster onset time for complete motor blockade and in a prolonged duration of motor effects. Liposomal formulation appears to be a powerful delivery system to prolong the motor effects of bupivacaine since E50 was lower and Emax higher than after the use of plain solution (E50 4.49+/-1.81 mg and Emax 152+/-40 min for P; and E50 2.61+/-0.23 mg and Emax 202+/-9 min for L). Hemodynamic changes were linearly related to doses of bupivacaine injected. The best bupivacaine-to-lipid ratio to prolong motor effects using our model was 3.75 mg and 20.0 mg respectively (Lipo 3.7). PMID- 10370176 TI - Stealth PEGylated polycyanoacrylate nanoparticles for intravenous administration and splenic targeting. AB - The aim of the present work was to investigate the biodistribution characteristics of PEG-coated polycyanoacrylate nanoparticles prepared by the nanoprecipitation/solvent diffusion method using the previously synthesized poly(MePEGcyanoacrylate-hexadecylcyanoacrylate) copolymer. It was observed that [14C]-radiolabeled PEGylated nanoparticles remained for a longer time in the blood circulation after intravenous administration to mice, compared to the non PEGylated poly(hexadecylcyanoacrylate) (PHDCA) nanoparticles. Furthermore, hepatic accumulation was dramatically reduced, whereas a highly increased spleen uptake was shown. The PEGylation degree of the polymer seemed not to affect the in vivo behavior of the nanoparticles, whereas previously obtained in vitro data have shown a modification of plasma protein adsorption depending on the density of PEG at the surface of the particles. Moreover, the study of the in vitro cytotoxicity of the nanoparticles revealed that the PEGylation of the cyanoacrylate polymer reduced its toxicity. These results open up interesting perspectives for the targeting of drugs to other tissues than the liver. PMID- 10370177 TI - Perivascular delivery of heparin for the reduction of smooth muscle cell proliferation after endothelial injury. AB - Thin flexible sheets composed of poly(lactic acid) (PLA) laminated polyanhydride, poly(erucic acid dimer-sebacic anhydride) (P(EAD-SA)), loaded with heparin were evaluated in vitro and in vivo. PLA was used for coating the polyanhydride to improve the release profile and improve the strength of the films. Heparin was released constantly for 20 days from PLA-coated 2% loaded P(EAD-SA). The uncoated film of P(EAD-SA) released heparin for only 4 days. The localized delivery of heparin around the carotid artery was investigated by implanting polymer loaded with [3H]heparin around the carotid artery of rats and the heparin release and tissue distribution was monitored. The maximum heparin concentration in the artery exposed to the drug was on day 4 for the P(EAD-SA) uncoated device (fast releasing system) and day 11 for the coated devices. The control artery, the uncovered segments of the artery, and the surrounding tissue contained negligible amounts of radioactivity. These data confirm that heparin was delivered locally without systemic exposure. Two independent animal studies were conducted to evaluate the effectiveness of these heparin-releasing devices. In both studies the balloon catheter injury in a rat model was used. After inflicting an injury to the common carotid, a matrix oriented with its long axis along the artery was placed under the injured portion of the vessel. In both studies the treated rats showed a very thin layer of neointima where the control group showed a significant reduction of the artery internal diameter with SMC neointima ratio greater than 1. PMID- 10370178 TI - Uncertainty quantification in the health consequences of the boiling liquid expanding vapour explosion phenomenon. AB - A methodology for estimating the risk owing to the phenomenon of boiling liquid expanding vapour explosion (BLEVE) in the presence of uncertainties both in the model and in the parameters of the models is presented. BLEVE takes place when a tank containing liquefied petroleum gas (LPG) is exposed to fire and fails catastrophically. Two models have been used in the estimation of the intensity of thermal radiation from the resulting fireball, namely the solid-flame model assuming an emission power independent of the combustion mass and the point source model that estimates the emissive power as a function of the combustion mass. Three measures of the BLEVE consequences, the intensity of thermal radiation, the dose of thermal radiation and the probability of loss of life as a result of the exposure to the thermal radiation and as a function of the distance from the center of the tank have been considered. Uncertainties in the exact values of the parameters of the models have been quantified and the resulting uncertainties in the three consequence measures have been assessed. A sensitivity analysis on the relative contribution of the uncertainty in each of the input variables to the uncertainties of the consequence measures has been performed. One conclusion is that the uncertainties in the probability of loss of life are mainly due to the uncertainties in the model of the physical phenomenon rather than to the uncertainties of the dose-response model. PMID- 10370180 TI - Ammonium nitrate: a promising rocket propellant oxidizer AB - Ammonium nitrate (AN) is extensively used in the area of fertilizers and explosives. It is present as the major component in most industrial explosives. Its use as an oxidizer in the area of propellants, however, is not as extensive as in explosive compositions or gas generators. With the growing demand for environmental friendly chlorine free propellants, many attempts have been made of late to investigate oxidizers producing innocuous combustion products. AN, unlike the widely used ammonium perchlorate, produces completely ecofriendly smokeless products. Besides, it is one of the cheapest and easily available compounds. However, its use in large rocket motors is restricted due to some of its adverse characteristics like hygroscopicity, near room temperature phase transformation involving a volume change, and low burning rate (BR) and energetics. The review is an attempt to consolidate the information available on the various issues pertaining to its use as a solid propellant oxidizer. Detailed discussions on the aspects relating to phase modifications, decomposition chemistry, and BR and energetics of AN-based propellants, are presented. To make the review more comprehensive brief descriptions of the history, manufacture, safety, physical and chemical properties and various other applications of the salt are also included. Copyright 1999 Elsevier Science B.V. PMID- 10370179 TI - Regulatory requirements for biocides on the market in the European Union according to Directive 98/8/EC. AB - In early 1998, the European Commission and Parliament adopted a new Directive concerning the placing on the market of biocidal products. The Directive is to be implemented in the member states by May 2000. The member states are currently concerned with the national implementation into legislation whereas the Commission is setting up the proposal for a review programme for the existing active substances and the products in which they are used. This paper describes the effort currently undertaken (up to the end of December 1998) to define the procedures to be used and characterise the substances covered. PMID- 10370181 TI - Geotechnical techniques for the construction of reactive barriers AB - One of the newest and most promising remediation techniques for the treatment of contaminated groundwater and soil is the reactive barrier wall (commonly known as PRB for permeable reactive barrier or reactive barrier). Although a variety of treatment media and strategies are available, the most common technique is to bury granular iron in a trench so that contaminated groundwater passes through the reactive materials, the contaminants are removed and the water becomes 'clean'. The principal advantages of the technique are the elimination of pumping, mass excavation, off-site disposal, and a very significant reduction in costs. The use of this technology is now becoming better known and implemented. Special construction considerations need to be made when planning the installation of reactive barriers or PRBs to ensure the design life of the installation and to be cost-effective. Geotechnical techniques such as slurry trenching, deep soil mixing, and grouting can be used to simplify and improve the installation of reactive materials relative to conventional trench and fill methods. These techniques make it possible to reduce the hazards to workers during installation, reduce waste and reduce costs for most installations. To date, most PRBs have been installed to shallow depths using construction methods such as open trenching and/or shored excavations. While these methods are usable, they are limited to shallow depths and more disruptive to the site's normal use. Geotechnical techniques are more quickly installed and less disruptive to site activities and thus more effective. Recently, laboratory studies and pilot projects have demonstrated that geotechnical techniques can be used successfully to install reactive barriers. This paper describes the factors that are important in designing a reactive barrier or PRB installation and discusses some of the potential problems and pitfalls that can be avoided with careful planning and the use of geotechnical techniques. Copyright 1999 Elsevier Science B.V. PMID- 10370182 TI - A new direct microscopy based method for evaluating in-situ bioremediation. AB - A new epifluorescent microscopy based method using 5-cyano-2, 3-ditolyl tetrazolium chloride (CTC) and 5-(4,6-dichlorotriazinyl) aminofluoroscein (DTAF) was developed for quantifying total microbial biomass and evaluating levels of microbial activity. CTC is a tetrazolium dye that forms fluorescent intracellular formazan when biologically reduced by components of the electron transport system and/or dehydrogenases of metabolically active bacteria. DTAF is a fluorescein based fluorochrome that selectively stains bacterial cell walls thereby enabling quantification of total bacterial biomass. CTC can be used in conjunction with DTAF to provide the optical resolution necessary to differentiate metabolically active cells from inactive cells in microbial populations associated with subsurface soils. The CTC/DTAF staining method has been shown to be effective for quantifying the metabolic activity of not only aerobic bacteria, but also diverse groups of anaerobic bacteria. This method allows for the rapid quantification of total and active bacterial numbers in complex soil samples without enrichment or cell elution. In this study, CTC/DTAF staining was applied to evaluate in-situ microbial activity in petroleum hydrocarbon contaminated subsurface soils from Sites 3 and 13 at Alameda Point, CA. At each site, subsurface microbial activity at two locations within contaminated plumes were examined and compared to activity at two geologically similar but uncontaminated background locations. Significant bacterial populations were detected in all soils examined, and the biomass estimates were several orders of magnitude higher than those obtained by conventional culture-based techniques. Both the total bacterial concentrations and the numbers of active bacteria in soils from contaminated areas were substantially higher than those observed in soils from background locations. Additionally, the percentages of metabolically active bacteria in the contaminated areas were consistently higher than those detected in background areas, suggesting that the enhanced microbial activity was due to microbial contaminant degradation. Although conventional heterotrophic plate counts failed to show significant microbial activity at either of the sites, soil gas carbon dioxide and methane measurements confirmed that hydrocarbon contaminant degradation was occurring in both areas. The CTC/DTAF staining protocol proved to be a rapid, reliable, and inexpensive method to evaluate the progress of in-situ bioremediation. PMID- 10370183 TI - Laboratory model of a petroleum migration barrier in arctic alaska AB - Child's Pad is a gravel construction pad that was contaminated with petroleum during oil-field service operations in Deadhorse, AK. As part of a remedial action plan, a buffer strip of uncontaminated sandy gravel was placed along certain sections of the pad boundary. A peroxygen formulation manufactured by Regenesis Copyright, sold as Oxygen Release Compound (ORC(R)), was placed in the buffer strips. The ORC was intended to supply oxygen to aerobic microorganisms capable of degrading petroleum. Tests were conducted in a 1/2 scale laboratory cell to determine the oxygen release characteristics of the ORC when subjected to expected subsurface flow rates of up to 0. 02 l/s (6.9 m/day). In laboratory tests, a zone of enhanced oxygen concentration was formed down-gradient from the ORC socks. Only during periods when the flow rate was less than 0.01-0.015 l/s (3. 5-5.2 m/day) was ORC-oxygen observed at monitoring points up-gradient or directly cross-gradient of the ORC. Conclusions from the laboratory study were that ORC may provide an aerobic zone in the Child's Pad barrier as far as 1 m directly down-gradient of the sock during periods of high flows (6.9 m/day). Copyright 1999 Elsevier Science B.V. PMID- 10370184 TI - Evaluation of transdermal iontophoresis of enoxacin from polymer formulations: in vitro skin permeation and in vivo microdialysis using Wistar rat as an animal model. AB - Polymers were used in vehicles to form hydrogel matrices in this study to evaluate the in vitro permeation and in vivo microdialysis of enoxacin. The highest transdermal delivery determined by area under flux-time curve (AUC) and intracutaneous enoxacin concentration were observed in methylcellulose (MC) and polyvinylpyrrolidone (PVP) hydrogels, respectively. To avoid the pH shift in vehicles during iontophoresis, buffer species were added to formulations to increase the buffer capacity. As expected, the permeability of enoxacin of anodal iontophoresis was larger than that of cathodal iontophoresis. Combination of benzalkonium chloride, a cationic surfactant as an enhancer, and iontophoresis exerted an enhancing effect for anionic enoxacin at pH 10.0. However, no effect or a negative effect was detected for cationic enoxacin in deionized water or pH 5.0 buffer, due to the shielding of the negative charge in the skin. The skin residue of enoxacin was slightly increased after the incorporation of Azone in PVP hydrogel. The result of in vivo microdialysis was in accordance with that of in vitro study. The effect of Azone on the intracutaneous enoxacin was more significant for in vivo microdialysis than in the in vitro study indicating the clinical feasibility of Azone for iontophoretic delivery. Microdialysis can be considered as a useful technique to investigate the pharmacokinetics of transdermal iontophoresis in vivo. PMID- 10370185 TI - Acyloxymethyl acidic drug derivatives: in vitro hydrolytic reactivity. AB - A series of acyloxymethyl drug derivatives of the NH-acidic drugs, phenytoin and theophylline and of the carboxylic acid drugs, thioctic acid and indomethacin, were prepared in order to determine the effect of varying the nature of the drug on the in vitro rate of hydrolysis catalyzed by porcine liver esterase and human plasma. The acyl portion was comprised of either valeric acid (val) or gamma linolenic acid (GLA). With the exception of some GLA prodrugs, the derivatives displayed first-order kinetics in both enzyme systems. The NH-acidic drug derivatives were hydrolyzed faster than the carboxylic drug derivatives by porcine liver esterase and human plasma. It was found that the short chain valeric acid derivatives were hydrolyzed faster than the GLA derivatives. The rates of hydrolysis for the relatively smaller prodrugs of theophylline and thioctic acid were greater than the rates of hydrolysis for the bulkier phenytoin and indomethacin prodrugs indicating steric hindrance was important. The lipophilicity index, log K, of the valeric acid drug derivatives was plotted against the logarithm of the hydrolysis rate constant, k, and it was observed that log k decreased with an increase in log K. A comparison of these results with those of previous studies where the alkyl and acyl moieties were varied of acyloxyalkyl theophylline derivatives has provided a rationale, based on lipophilicity, for the structure of a prodrug to be designed based on an in vitro desired rate of hydrolysis. PMID- 10370186 TI - Investigations into the structure and composition of beta cyclodextrin/poly(acrylic acid) microspheres. AB - Microspheres composed of the hydrophilic polymer poly(acrylic acid) (PAA), with and without beta-cyclodextrin (beta-CD), were prepared by a water-in-oil (w/o) solvent evaporation technique. Microspheres were characterised for particle size, beta-CD and residual oil content. The type of matrix formed during microsphere synthesis was investigated by solid state carbon 13C NMR, in vitro release of beta-CD and swelling measurements. A high encapsulation efficiency of the beta-CD was observed (?90%). The in vitro release of beta-CD in water over 24 h was initially rapid ( approximately 70% in 3 h) with no further loss thereafter, suggesting potential covalent binding of the residual beta-CD. NMR indicated that in the presence of beta-CD, two concomitant chemical processes occur during microsphere synthesis: (i) esterification of the hydroxyl group(s) of the beta-CD with the carboxylic acid groups of the PAA; and (ii) the formation of intra /inter-polymer acid anhydrides. PMID- 10370187 TI - A comparison of the bronchodilatory effect of 50 and 100 microg salbutamol via Turbuhaler and 100 microg salbutamol via pressurized metered dose inhaler in children with stable asthma. AB - AIM: The aim of the study was to compare the efficacy of single doses of salbutamol Turbuhaler (50 and 100 microg), salbutamol pressurized metered dose inhaler (pMDI) (100 microg) and placebo in children with stable chronic reversible airway obstruction. Primary efficacy variable (FEV1-av) was calculated as the area under the curve of forced expiratory volume in one second (FEV1) (AUC, 0-4 h) and divided by the observed time. DESIGN: The study was of a randomized, single-dose, crossover and double-blind design. Seven centres participated. FEV1 was measured pre-dose and at 15 min, 0.5, 1, 1.5, 2, 3 and 4 h post study dose. PATIENTS: Forty asthmatic children (9 girls) with a mean age of 9 years (range: 6-12), mean FEV1 of 1.6 l (range: 0.9-2.4) and a mean FEV1 in percentage of predicted normal value of 80% (range: 61-109) were randomized into the study. The mean reversibility 30 min after inhaling 2x100 microg salbutamol from pMDI was 20% (range: 9-45) or 15% (range: 8-27) in percentage of predicted normal value. RESULTS: The mean FEV1-av was 1.63 l for placebo, 1.71 l for 50 microg salbutamol Turbuhaler, 1.76 l for 100 microg salbutamol Turbuhaler and 1.76 for 100 microg salbutamol pMDI. Corresponding values for maximum FEV1 were 1.76, 1. 85, 1.87 and 1.87 l, respectively. There were no statistically significant differences between the active treatments in FEV1-av or maximum FEV1. All active treatments were significantly better than placebo. CONCLUSION: No significant differences in bronchodilating effect between 50, 100 microg salbutamol Turbuhaler and 100 microg salbutamol pMDI in children, aged 6-12 years, with stable asthma could be demonstrated. All active treatments were significantly better than placebo. PMID- 10370188 TI - Development of a membrane-controlled transdermal therapeutic system containing isosorbide dinitrate. AB - The formulation of a transdermal delivery system for isosorbide dinitrate (ISDN) was examined. It was found that the target release rate should be 4.01 mg/h per 20 cm2 for optimal dosing. In order to reach such this zero order release rate, a membrane permeation controlled transdermal therapeutic system (TTS) formulation was developed, with ethylene vinyl acetate copolymer (EVAC) and polyethylene (PE) membranes as rate controlling membranes; a carbomer gel was used as the drug reservoir. The release of ISDN from this drug delivery device was studied in vitro using FDA recommended method. PIB adhesive on the EVAC or PE membrane caused a decreased flux of ISDN; the release kinetics fitted Higuchi matrix kinetics. TTS with EVAC membrane release ISDN at a rate much lower than the calculated target release rate, but with PE membranes, the release rate was very close to the target. Release rate studies have revealed that, as the VA content in EVAC membrane increased, the flux of ISDN increased. All these results were compared with the commercial product Frandol(R) Tape S from Japan. It was found that the release rate of Frandol was close to target release rate and fitted matrix kinetics. These results suggested that TTS that contain PE membrane as rate controlling membrane, polyisobutylene (PIB) adhesive and carbomer gel as a reservoir can be applicable as a TTS for ISDN. PMID- 10370189 TI - Topical use of chitosan in ophthalmology: tolerance assessment and evaluation of precorneal retention. AB - The mucoadhesive polysaccharide chitosan was evaluated as a potential component in ophthalmic gels for enabling increased precorneal drug residence times. This cationic vehicle was expected to slow down drug elimination by the lacrymal flow both by increasing solution viscosity and by interacting with the negative charges of the mucus. The molecular weight (Mw) and concentration of polysaccharide were studied in four types of chitosan as parameters that might influence ocular tolerability and precorneal residence time of formulations containing tobramycin as therapeutic agent. An ocular irritation test, using confocal laser scanning ophthalmoscopy (CLSO) combined with corneal fluorescein staining, clearly demonstrated the excellent tolerance of chitosan after topical administration onto the corneal surface. Gamma scintigraphic data showed that the clearance of the formulations labelled with 99mTc-DTPA was significantly delayed in the presence of chitosan with respect to the commercial collyrium (Tobrex(R)), regardless of the concentration and of the molecular weight of chitosan in solution. At least a 3-fold increase of the corneal residence time was achieved in the presence of chitosan when compared to Tobrex(R). PMID- 10370190 TI - Microagglomeration of pulverized pharmaceutical powders using the Wurster process I. Preparation of highly drug-incorporated, subsieve-sized core particles for subsequent microencapsulation by film-coating. AB - A novel agglomeration process of pulverized pharmaceutical powders into subsieve sized agglomerates (microagglomeration) was designed for manufacturing highly drug-incorporated core particles for subsequent microencapsulation by film coating. The microagglomeration of pulverized phenacetin powder, whose mass median diameter was 9 microm, was performed by spraying an aqueous colloidal dispersion of acrylic polymer, Eudragit(R) RS30D, as a binding/coating agent using a spouted bed assisted with a draft tube (the Wurster process), and the effect of process variables was examined. An appropriate spray liquid flow rate made it possible to produce microagglomerates of 20-50 microm with 60% yield. However, 10% of the product still survived as particles smaller than 10 microm even at the elevated liquid flow rate. In contrast, the survived particles smaller than 10 microm tended to be predominantly reduced to 2%, while coarse agglomerates larger than 53 microm were not excessively produced, by additionally setting a fixed bed of glass beads in the spouted bed apparatus. The length of the draft tube influenced compaction of the agglomerates as well as their surface smoothening. Equipping the fixed bed of the glass beads and the long draft tube in the spouted bed allowed us to prepare microagglomerates of 20-50 microm at yield of 55% applicable as highly drug-incorporated, free-flowing, surface smoothed, narrowly size-distributed core particles for subsequent microencapsulation by film-coating. PMID- 10370191 TI - Interaction of a nonionic surfactant-based organogel with aqueous media. AB - In an attempt to explain the rather short half-life of molecules at the injection site after their intra-muscular administration in a sorbitan monostearate organogel, in vitro studies were carried out to study the effects of an aqueous medium (simulating interstitial fluid at injection site) on the physical form of the organogel. When the gel mass comes in contact with an aqueous phase, the latter penetrates into the organic gel via the sorbitan monostearate tubular network, resulting in gel breakdown into smaller fragments. The surfactant tubular network act as a conduit for water penetration into the gel. Meanwhile, emulsification, aided by the surfactants present in the gel, also occurs at the gel surface between the organogel and the aqueous phase. This leads to a gradual erosion of the gel as oil droplets bud off from the gel mass. From these in vitro observations, we speculate that after gel administration in vivo, dynamic interactions occur between the local interstitial fluid and the gel mass: fluid penetration into the gel and emulsification at the gel surface is thus responsible for gel breakdown and so a relatively short duration of drug at the injection site. PMID- 10370192 TI - Methyl-beta-cyclodextrin and doxorubicin pharmacokinetics and tissue concentrations following bolus injection of these drugs alone or together in the rabbit. AB - The purpose of this work was to determine the pharmacokinetics and the tissue concentrations of methyl-beta-cyclodextrin (MEBCD) and doxorubicin (DOX) in rabbits following administration of MEBCD and DOX, alone or in combination. MEBCD (200 mg/kg) and DOX (1 mg/kg) were intravenously injected to white New Zealand rabbits and blood samples were obtained over a 48-h period after administration. After this period, administration was repeated and animals were killed 1, 2 or 4 h after injection. Heart, liver and kidney were then removed. MEBCD and DOX analysis in plasma and tissues was performed using two HPLC methods with fluorimetric detection. MEBCD pharmacokinetic profile was consistent with a two compartment model (t1/2 alpha: 30 min; t1/2 beta: 7 h). Co-administration with DOX did not modify the main pharmacokinetic parameters of MEBCD. However, C5 min, t1/2 alpha, t1/2 beta and AUCinfinity were decreased by the co-administration of DOX with MEBCD compared to DOX alone. Assays of excised tissues showed that DOX enhanced the cardiac, renal and hepatic concentrations of MEBCD. On the other hand, MEBCD did not alter the cardiac distribution of DOX, while renal and hepatic distribution profiles were modified. In this study, the pharmacokinetic parameters of MEBCD injected intravenously were determined for the first time. DOX did not enhance MEBCD pharmacokinetic profile but MEBCD reduced the distribution half-life of DOX. Tissue determination showed that MEBCD did not enhanced the cardiac accumulation of DOX, which is auspicious for further in vivo experiments using the co-administration of DOX and MEBCD. PMID- 10370193 TI - Temporal dependence of ectopeptidase expression in alveolar epithelial cell culture: implications for study of peptide absorption. AB - There is little data available regarding the extent of peptide metabolism encountered following inhalation to the lung. We have studied the activity of five ectopeptidases in primary rat alveolar epithelial cells, A549 cells and pulmonary macrophages. Peptidase activity was assayed in the plasma membrane fractions (PMF) of primary type II alveolar epithelial cells (ATII cells) after 2 days in culture and after 7 days in culture when they had formed monolayers of type I-like cells (ATI-like cells). Dipeptidyl peptidase IV (DPP) activity fell from 36.65 mU/mg protein to 16.29 mU/mg protein between day 2 and day 7 in culture, aminopeptidase N (AMN) activity increased from 16.16 mU/mg protein to 23.99 mU/mg protein, angiotensin-converting enzyme (ACE) activity was lost (4.29 mU/mg protein at day 2, not detected at day 7), and carboxypeptidase M (CPM) activity was acquired (not detected at day 2, 5.20 mU/mg protein at day 7). The profile of exopeptidase expression in A549 cells was similar to that of primary rat alveolar cells at day 7 in culture (DPP 24.24 mU/mg protein, AMN 47.74 mU/mg protein, CPM 4.28 mU/mg protein, ACE not detected). Macrophages expressed high levels of aminopeptidases (DPP 46.85 mU/mg protein, AMN 28.28 mU/mg protein) but carboxypeptidase activity was not detected. Low neutral endopeptidase 24.11 (NEP) activity was found in all cell types studied (0.96-2.41 mU/mg protein). The qualitative and quantitative changes in the peptidase activity of primary cultured rat alveolar cells between day 2 and day 7 in culture has implications for the use of alveolar cell monolayers as drug absorption models to investigate peptide absorption from the lung. Ectopeptidase activity in cultured alveolar cells can be used to infer the peptide-metabolising capacity of the surface of the alveolar epithelium. The aminopeptidase activity in particular, if representative of enzyme activity in vivo, would offer a significant metabolic barrier to systemic delivery of peptides via the lung. PMID- 10370194 TI - Effect of linolenic acid/ethanol or limonene/ethanol and iontophoresis on the in vitro percutaneous absorption of LHRH and ultrastructure of human epidermis. AB - The effect of enhancer(s) (e.g. ethanol (EtOH), 5% linolenic acid/EtOH, and 5% limonene/EtOH) and iontophoresis was investigated on the in vitro percutaneous absorption of luteinizing hormone releasing hormone (LHRH) and ultrastructure of human epidermis by transmission electron microscopy (TEM). 5% linolenic acid/EtOH or 5% limonene/EtOH significantly enhanced (P<0.05) the passive flux of LHRH through human epidermis in comparison to the control (no enhancer treated epidermis). Iontophoresis further increased the flux of LHRH through enhancer(s) treated epidermis. Iontophoretic flux of LHRH through 5% linolenic acid/EtOH and 5% limonene/EtOH treated epidermis was significantly (P<0.05) enhanced in comparison to iontophoretic flux through the control epidermis. TEM is the most efficient way to visualize the ultrastructure of the stratum corneum (SC). TEM results reveal that iontophoresis in combination with enhancers (e.g. linolenic acid/EtOH or and limonene/EtOH) transformed the highly compact cells of the SC into a looser network of filaments, disrupted the keratin pattern, and resulted in swelling of SC cell layers of human epidermis. Thus, linolenic acid/EtOH or limonene/EtOH in combination with iontophoresis increased the flux of LHRH through human epidermis by disrupting keratin pattern as well as loosening and swelling of SC cell layers. PMID- 10370195 TI - Toxicity of a particulate formulation for the intraperitoneal application of mitoxantrone. AB - Mitoxantrone (MXN) has demonstrated therapeutic efficacy in the intraperitoneal treatment of malignancies. However, severe local toxicity is dose limiting. Therefore, a particulate formulation of MXN, the drug incorporated in albumin microspheres, was evaluated concerning tolerability. Survival rates as well as alterations in body weight, food intake, water intake, urine volume, urine specific gravity, urine protein content, and complete blood count were observed following single or multiple intraperitoneal injections of MXN solution, dispersions containing MXN-loaded microspheres or unloaded microspheres, and the injection vehicle to female and male Sprague-Dawley rats. Applied MXN dosage was equivalent to 30 mg/m2 body surface area. Unloaded microspheres were well tolerated without signs of toxicity. Application of MXN solution or MXN-loaded microspheres resulted in similar survival rates (56% 9 weeks after single injection) and in a comparable bone marrow toxicity (mainly leucopenia). Body weight, food and water intake as well as urine volume were decreased following application of MXN solution, whereas a progressive gain in weight and no remarkable alterations in nutrition and urine excretion were noted after administration of MXN-loaded and unloaded microspheres, or of the injection vehicle. In conclusion, intraperitoneal injection of MXN incorporated in albumin microspheres exhibits in part less toxicity than conventional treatment. PMID- 10370196 TI - Liposomes as carriers of the antiretroviral agent dideoxycytidine-5' triphosphate. AB - The presence and replication of the human immunodeficiency virus (HIV) in cells of the mononuclear phagocyte system (MPS) together with the preferential uptake of liposomes in macrophages suggest that liposomes can become a valuable carrier of anti-HIV agents. Moreover, liposomes reduce toxicity of encapsulated drugs and protect encapsulated drugs against rapid degradation in the blood circulation. To overcome problems associated with the administration of free nucleosides and to improve targeting to the MPS, dideoxycytidine-5'-triphosphate (ddCTP) was encapsulated in liposomes. Liposomes were stable with regard to retention of the entrapped drug, particle size and chemical stability of ddCTP. Results obtained with liposome encapsulated ddCTP in the murine acquired immunodeficiency syndrome (MAIDS) model indicate that ddCTP encapsulated in liposomes can reduce proviral DNA in cells of the mononuclear phagocyte system (MPS) in both spleen and bone marrow. PMID- 10370197 TI - Pulmonary distribution and clearance of two beclomethasone liposome formulations in healthy volunteers. AB - The pulmonary distribution and clearance of 99mTc-labelled beclomethasone dipropionate (Bec) dilauroylphosphatidylcholine (DLPC) and dipalmitoylphosphatidylcholine (DPPC) liposomes were compared in 11 healthy volunteers using gamma scintigraphy. As delivered by using the Aerotech jet nebulizer both liposome aerosols had a suitable droplet size (mass median aerodynamic diameter 1.3 microm) allowing deep pulmonary deposition. However, in the total drug output during the inhalation there was a relatively large difference between DLPC and DPPC of 11.4 and 3.1 microg, respectively. In a gamma camera study no significant differences existed in the central/peripheral lung deposition between the DLPC and DPPC formulations. Progressive clearance of both Tc-labelled Bec liposomes was seen: 24 h after inhalation, 79% of the originally deposited radioactivity of DLPC liposomes and 83% of that of DPPC liposomes remained in the lungs. Thus there was slightly slower clearance of inhaled liposomes using DPPC instead of DLPC. We conclude that both liposome formulations are suitable for nebulization, although aerosol clouds were more efficiently made from the DLPC liposome suspension. Our results support the view that liposome encapsulation of a drug can offer sustained release and drug action in the lower airways. PMID- 10370198 TI - Dehydration and rehydration of a hydrate diclofenac salt at room temperature. AB - The salt diclofenac/N-(2-hydroxyethyl) pyrrolidine crystallizes from water as a dihydrate, while it precipitates from organic solvents anhydrously: the two salts have different crystal structures. Dehydration of the dihydrate salt was carried out in a desiccator over silica gel at room temperature: the process occurs with the retention of the crystal structure. Slight changes observed in the diffractograms suggest, that soon after dehydration, a phase transition starts, slowly due to the low temperature of the process. The reaction was followed determining the loss of weight as a function of time and by thermal analysis, since the dihydrate and the dehydrate forms have different thermograms, but similar diffractograms. The reaction was complete after 24 h. The analysis of the experimental data suggests a kinetic process related to a one-dimensional diffusion of the crystallization water molecules outwards the solid particles. At room temperature, the dehydrate material rapidly back-absorbs the two molecules of crystallization water from the atmosphere moisture. The interaction with water of the different forms of the salt was discussed as a function of their solid structures as well as of the complex equilibria present in aqueous solution: these can explain previous apparently anomalous results. PMID- 10370199 TI - Design of controlled release inert matrices of naltrexone hydrochloride based on percolation concepts. AB - The percolation theory is a statistical theory able to study chaotic or disordered systems that has been applied in the pharmaceutical field since 1987. Through the application of this theory, the design of controlled release inert matrices has been improved. The aim of the present paper is to estimate the percolation thresholds, the most important concept of the percolation theory, which characterise the release behaviour of controlled release inert matrices of naltrexone hydrochloride. Matrix tablets were prepared using naltrexone hydrochloride as a potent narcotic antagonist and Eudragit(R) RS-PM as matrix forming material in different ratios, keeping constant the drug and excipient particle sizes. In vitro release assays were carried out exposing only one side of the tablets to the dissolution medium. The drug percolation threshold was estimated using different methods. The method of Leuenberger and Bonny gives 31.11+/-7.95% v/v as the critical porosity, which corresponds to a percolation range from 12 to 20% (w/w) of drug content. The release profiles and the release kinetics are in agreement with this result. A change in the exponent k (from 0.29 to 0.57) has been found in this region. Using scanning electron microscopy, the percolation threshold has been observed in a higher concentration range (20-35% w/w). This fact can be attributed to the low accuracy of the visual methods, mainly due to the extrapolation from 2D to 3D systems. If a percolating cluster is observed in two dimensions, the percolation threshold of the 3D system will be already clearly exceeded. The excipient percolation threshold is estimated between 25.4 and 31.1% (v/v) based on the release profiles and the analysis of the release kinetics. PMID- 10370200 TI - Degradation kinetics of 4-dedimethylamino sancycline, a new anti-tumor agent, in aqueous solutions. AB - The kinetics of degradation of the new anti-tumor drug, 4-dedimethylamino sancycline (col-3) in aqueous solution at 25oC were investigated by high-pressure liquid chromatography (HPLC) over the pH-range of 2-10. The influences of pH, buffer concentration, light, temperature, and some additives on the degradation rate were studied. The degradation of col-3 was found to follow first order kinetics. A rate expression covering the degradation of the various ionic forms of the drug was derived and shown to account for the shape of the experimental pH rate profile. Under basic conditions, the degradation of col-3 involves oxidation, which is catalyzed by metal ions and inhibited by EDTA and Sodium bisulfite. PMID- 10370201 TI - Water sorption and near IR spectroscopy to study the differences between microcrystalline cellulose and silicified microcrystalline cellulose before and after wet granulation. AB - Silicified microcrystalline cellulose (SMCC) has been shown to have advantages over conventional microcrystalline cellulose (MCC). These advantages are (i) improved tablet strength compared to that achieved with MCC, (ii) the retention of compressibility after wet granulation, whereas MCC produces weaker tablets after wet granulation, and (iii) superior flow properties than MCC. In this study gravimetric and calorimetric vapour sorption data and near IR spectroscopy have been used to study MCC and SMCC before and after wet granulation. It was found that MCC, SMCC and wet granulated SMCC had essentially identical physical structures (except for a size increase due to granulation). Wet granulated MCC had a different enthalpy of water sorption at low RH, and its near IR spectrum was different from the other samples in the region which relates to C-H bonding. It can be concluded that MCC and SMCC are of very similar structures, thus these analytical techniques cannot provide an explanation for the improvements in compressibility. However the change in compressibility in MCC after wet granulation may relate to the observed differences in internal bonding in this sample. PMID- 10370202 TI - Chitinosans as tableting excipients for modified release delivery systems. AB - The term 'chitinosans' embraces the spectrum of acetylated poly(N-glucosamines) ranging from chitin to chitosan. Chitinosans (I), at acidic pH, have protonated amines which can interact with oppositely charged drug ions and, thereby, modify drug release from drug delivery systems. Tablets were compressed from a physical mixture containing salicylic acid (II) as the model drug, I, and magnesium stearate. Five commercial I compounds, varying in degree of deacetylation and molecular weight, were selected. Tablets were compressed at 5000, 10 000, and 15 000 psig using a Carver and a single punch tablet press. The differential scanning calorimetry thermograms provided evidence of I-II interaction in the powder blend. Analysis of variance (ANOVA) indicated that the compression pressure did not significantly affect the crushing strength (CS) or the release profile of II from the I-matrix tablets (P?0.05). Furthermore, the ANOVA also indicated that the tablet press used during manufacture did not affect the above properties (P?0.05); however, the chitinosans significantly affected the CS as well as the release profile of II from I-matrix tablets (P<0.05). This study provides further evidence for the use of commercial I compounds as excipients for use in modified release drug delivery systems. PMID- 10370203 TI - Incorporation mechanism of guest molecules in crystals: solid solution or inclusion? AB - Guest molecules (impurities or additives), together with some crystallization solvent, are often incorporated into the host crystals during crystallization from solution. The guest molecules may be incorporated either in solid solution or in liquid inclusions, or by both mechanisms. The mechanism of guest incorporation has been examined by a simple calculation method which is based on the equality of the guest/solvent mole ratio in the initial crystallization medium and in the putative inclusions. Application of this calculation method to eight guest+host systems described in the literature has shown that a negligible amount (at most 0.2%) of the guest molecules is incorporated into the crystal lattice in liquid inclusions. Therefore, it is concluded that the vast majority of the guest molecules are incorporated into the crystals in solid solution, as previously suggested, but hitherto unproven, for these guest-host systems. PMID- 10370204 TI - Ocular devices for the controlled systemic delivery of insulin: in vitro and in vivo dissolution. AB - Both in vitro flow-through and in vivo device removal methods were utilized to determine the dissolution rate of insulin from a Gelfoam(R) based eye device. The dissolution profiles generated by these two methods are comparable. The in vivo data suggests that there is a direct relationship between blood glucose lowering and the rate of release of insulin from the device. The in vitro dissolution results indicate that the release of insulin from the device is flow-rate dependent. The prolonged activity of the insulin is due to the gradual release of insulin from the device which results from the lachrymal system's slow and constant tear production. PMID- 10370205 TI - Targetability of novel immunoliposomes prepared by a new antibody conjugation technique. AB - In order to develop long-circulating immunoliposomes (IL), which combine sterical stabilization with a superior targetability, we have introduced a new methodology for attaching monoclonal antibodies directly onto the distal ends of liposome grafted polyethylene glycol (PEG) chains. Therefore, we have synthesized a new PEG-PE derivative, which had been endgroup-functionalized with cyanuric chloride. Antibodies can simply be coupled to this membrane anchor in mild basic conditions (pH 8.8) without the need for previous antibody derivatizations. The coupling results have been determined with consideration to various liposome parameters and have been compared to several established antibody coupling procedures, where antibodies had been linked directly to the liposome surface in the presence of PEG (conventional IL). To investigate the targetability of the resulting new IL, anti E-selectin mAb have been coupled and the degree of binding selectin containing cells has been analyzed. The terminal coupled antibodies show a 1.8 fold higher degree of in vitro cell binding compared to conventional IL, which has been attributed to the antibody position being more easy accessible at the PEG termini. Furthermore, we have illustrated the liposome surface topology and the coupled antibodies by atomic force microscopy, which for such fluid IL has been used first. These images have finely corresponded to the cell binding results, and have been discussed in terms of antibody position and flexibility at the liposome surface. PMID- 10370206 TI - Effect of the temperature on a hydrate diclofenac salt. AB - The salt Diclofenac/N-(2-hydroxyethyl) pyrrolidine when crystallizes from water forms a di-hydrate, which looses the crystallization water molecules on heating or in the presence of silica gel, undergoing a phase transition. The two processes were followed at room temperature, at 40 and 50 degrees C by thermal analysis and analyzing the dimensional parameters obtained by scanning electron microscopy as a function of the changes occurring in the solid state. The fractal dimension of the particle surface (DS) was determined for the di-hydrate, the anhydrate and the anhydrous forms: DS values are close together suggesting that the processes modify only slightly the external morphology of the particles. The reactive dimension (DR) to dissolution suggests that the salt after the thermal treatment has a dissolution behaviour identical to that observed for the salt obtained from organic solvents. The two processes de-hydration and phase transition can be carried out at relatively low temperature, suggesting an important pathway to obtain the anhydrous form starting from the di-hydrate one. PMID- 10370207 TI - Release of adriamycin from poly(gamma-benzyl-L-glutamate)/poly(ethylene oxide) nanoparticles. AB - Prolonged circulation of anticancer agent in blood is expected to decrease the host toxicity and enhance the anticancer activity. The purpose of this study is to develop and characterize the prolonged and sustained release formulation of anticancer agent using biodegradable poly(gamma-benzyl-L-glutamate)/poly(ethylene oxide) (PBLG/PEO) polymer nanoparticles. PBLG/PEO polymer is a hydrophilic/hydrophobic block copolymer and forms a micelle-like structure in solution. Spherical nanoparticles incorporating adriamycin were prepared by a dialysis method. The fluorescence intensity of adriamycin in the nanoparticles was increased when sodium dodecylsulfate was added. It is one of the evidences of entrapment of adriamycin in the polymer nanoparticles. Only 20% of entrapped drug was released in 24 h at 37 degrees C a and the release was dependent on the molecular weight of hydrophobic polymer. The endothermic peak of adriamycin at 197 degrees C disappeared in the nanoparticles system, suggesting the inhibition of a crystallization of adriamycin by polymer adsorption during the precipitation process. The mean residence time of adriamycin from the nanoparticles was more than threefold that from a free adriamycin. These results suggest usefulness of PBLG/PEO nanoparticles as a sustained and prolonged release carrier for adriamycin. PMID- 10370208 TI - The effect of daunorubicin and glutaraldehyde treatment on the structure of erythrocyte membrane. AB - The effect of daunorubicin (DNR) and glutaraldehyde on erythrocyte membrane structure was examined by Electron Spin Resonance spectroscopy. Human erythrocytes were incubated with daunorubicin and then with glutaraldehyde to prevent drug efflux. We have demonstrated that DNR alone caused changes in membrane fluidity mainly in the hydrophobic regions of the lipid bilayer. When DNR-preincubated erythrocytes were treated with glutaraldehyde, the alterations in fluidity were observed in the polar regions as well as in the deeper regions of the cell membrane. The incorporation of drug and glutaraldehyde into human erythrocytes also caused conformational alterations in membrane cytoskeletal proteins and changes in the internal viscosity of the cells. The results suggest that glutaraldehyde in the drug-pretreated erythrocytes may lead to significant perturbations in the organization of the plasma membrane lipids and proteins. PMID- 10370209 TI - Nasal mucoadhesive delivery systems of the anti-parkinsonian drug, apomorphine: influence of drug-loading on in vitro and in vivo release in rabbits. AB - Lyophilized polyacrylic acid powder formulations loaded with apomorphine HCl were prepared and the influence of drug loading on in vitro release and in vivo absorption studied after intranasal administration in rabbits. These formulations prepared with Carbopol 971P, Carbopol 974P and polycarbophil sustained apomorphine release both in vitro and in vivo. The in vitro release rate and mechanism were both influenced by the drug loading. There was no large influence of drug loading on the time to achieve the peak (Tmax) for a particular polymer, but Tmax differed between different polymers. For a particular drug loading, the Tmax from Carbopol 971P was the slowest compared with that for Carbopol 974P and polycarbophil; however, only the Tmax from Carbopol 971P loaded with 15% w/w of apomorphine was significantly longer than polycarbophil of similar drug loading (P=0.0386). The trend further observed was that increasing drug loading led to increased peak plasma concentration and area under the curve (AUC). In the second part of this study, a mixture containing an immediate release component and sustained release formulation was administered in an attempt to increase the initial plasma level, as this could be therapeutically beneficial. Only one peak plasma concentration was observed and the initial plasma concentrations were no higher than those obtained with solely sustained release formulation. The Tmax, the peak plasma drug concentration (Cmax) and AUC from the lactose-containing formulation were lower than the formulation without lactose but the differences were only marginally statistically significant for Cmax (P=0.0911) and AUC (P=0.0668), but not Tmax (P=0.2788). PMID- 10370210 TI - Properties of solid dispersions of piroxicam in polyvinylpyrrolidone. AB - Solid dispersions of piroxicam were prepared with polyvinylpyrrolidone (PVP) K-17 PF and PVP K-90 by solvent method. The physical state and drug:PVP interaction of solid dispersions and physical mixtures were characterized by X-ray diffraction, Fourier transform infrared (FTIR) spectroscopy and differential scanning calorimetry (DSC). FTIR analysis demonstrated the presence of intermolecular hydrogen bonding between piroxicam and PVP in solid dispersions. These interactions reflected the changes in crystalline structures of piroxicam. The amorphousness within the PVP moeity might be predicted in piroxicam dispersions by the disappearance of N-H or O-H peak of piroxicam. Dissolution studies indicated a significant increase in dissolution of piroxicam when dispersed in PVP. The better results were obtained with the lower molecular weight PVP K-17 than with higher molecular weight PVP K-90. The non-amorphous solid dispersions in PVP K-17 showed almost equally fast dissolution rates to amorphous dispersions in PVP K-90. The mechanism of dissolution of solid dispersion in PVP K-90 is predominantly diffusion-controlled due to the very high viscosity of PVP K-90. Dissolution was maximum with the amorphous solid dispersions containing drug:PVP K-17 1:5 and 1:6 which showed a 40-fold increase in dissolution in 5 min as compared with pure drug. PMID- 10370211 TI - Modeling of supersaturated dissolution data. AB - A recursion equation which relies on the population growth model of dissolution is used for the analysis of supersaturated dissolution data. The concentration time data of dissolution experiments are initially transformed to fractions of dose dissolved-generations by adopting an appropriate time interval as the time step of the recursion equation. A computer program is used to derive estimates for the maximum fraction of dose dissolved and the fraction of dose remaining in solution at steady state. Good fittings were observed when this equation was applied to phenytoin and nifedipine supersaturated dissolution data obtained from literature. PMID- 10370212 TI - Protein encapsulation in biodegradable amphiphilic microspheres. AB - MPOE-PLA microspheres containing bovine serum albumin (BSA) were prepared by the double emulsion method with high encapsulation efficiency ( approximately 93%). Confocal scanning microscopic analysis using MPOE-PLA labelled with 1 pyrenemethanol showed the MPOE coating of the microsphere surface. This coating improves the performance of the release system compared with PLA microspheres; the hydrophilic chains reduce the BSA adsorption onto the microspheres and increase the amount of BSA released in the supernatant. Microsphere analysis using atomic force microscopy showed that the presence of the MPOE chains also leads to surface roughness. Studies of the diffusion of 1% rhodamine aqueous solution into the microspheres by means of confocal microscopy showed a fast diffusion of water through the matrices containing high molecular weight MPOE chains (?10 000 g mol-1) and could explain the fast release of BSA from these microspheres. PMID- 10370213 TI - Preparation and evaluation of flurbiprofen-loaded microemulsion for parenteral delivery. AB - The purpose of this study was to improve the solubility of flurbiprofen, a poorly water-soluble drug, in an oil-in-water (o/w) microemulsion that is suitable for parenteral administration. Microemulsions with varying ratios of oil to surfactant were prepared with ethyl oleate, Tween 20 and isotonic solution. The effect of formulation variables on the particle size of microemulsion and solubility of flurbiprofen in microemulsion system was investigated. The pharmacokinetic parameters of flurbiprofen after intravenous administration of flurbiprofen-loaded microemulsion were compared with those of a solution of the drug. The mean droplet diameter of microemulsion containing less than 1% (w/w) of flurbiprofen was below 100 nm. The maximum solubility of flurbiprofen in the microemulsion system was found to be 10 mg/ml. However, the mean droplet diameters of flurbiprofen-loaded o/w microemulsions tend to be increased at room temperature. The pharmacokinetic parameters of flurbiprofen after intravenous administration of flurbiprofen-loaded microemulsion to rats were not significantly different from those of flurbiprofen in phosphate-buffered saline solution. It can be concluded that microemulsions of flurbiprofen prepared with ethyl oleate and Tween 20 can be used as a parenteral drug carrier for this and other poorly water-soluble drugs, provided that physical stability can be properly addressed. PMID- 10370214 TI - In vitro evaluation of biodegradable epsilon-caprolactone-co-D, L-lactide/silica xerogel composites containing toremifene citrate. AB - Poly(epsilon-caprolactone-co-D,L-lactide) polymers were blended with toremifene citrate or with toremifene citrate impregnated silica xerogel in order to develop a controlled release formulation. The copolymers were synthesized by bulk polymerization and characterized by nuclear magnetic resonance, size exclusion chromatography and differential scanning calorimetry analyses. The in vitro release of toremifene citrate, an antiestrogenic compound, and silica was carried out in simulated body fluid (pH 7.4) containing 0.5 wt% sodium dodecylsulphate at 34 degrees C. The in vitro release studies indicate that the release flux of toremifene citrate increases with increasing weight fraction of caprolactone in the copolymer. Silica xerogel had a minor enhancing effect on the release rate of toremifene citrate. Copolymers containing larger amounts of D,L-lactide (PLA-CL20 and PLA-CL40 copolymers) were not suitable matrices for the delivery of toremifene citrate in a controlled manner because of the burst effect. The fraction of toremifene citrate released from PLA-CL80 matrix increased with the increasing loading of toremifene citrate. The results of the study indicate that the in vitro release of toremifene citrate can be adjusted by varying the polymer composition and also the initial drug loading. PMID- 10370215 TI - Cerium-doped diosmectite for topical application studies of the cerium-clay interaction. AB - Cerium is used for its antiseptic and immunomodulatory properties in burn injury. We have developed a cerium-doped clay to replace existing ointments. Adsorption and release of cerium (Ce3+) by diosmectite were studied at 22+/-2 degrees C, in the presence of various other cationic species. Simple spectrofluorimetric determination of cerium was used (lambdaexc=240 nm/lambdaem=360 nm). Cerium binding reached a plateau within 2 min and was a function of the electrolyte content of the solution in contact with the clay. Langmuir isotherm treatment led to a maximal binding capacity of 66 mg of Ce3+ per gram of clay. Partial release occurred within 2 min (19% in the presence of isotonic NaCl solution). The ionic strength of the solution, and the ionic radius and charge of the electrolytes present in the bathing solution significantly influenced cerium release, in contrast to pH and temperature changes. These results strongly point to a cationic exchange mechanism between diosmectite and cerium solution. PMID- 10370216 TI - Transepithelial transport of bepridil in the human intestinal cell line, Caco-2, using two media, DMEMc and HBSS. AB - The purpose of this work was to study transepithelial transport of bepridil, an anticalcic agent, through monolayer cells Caco-2, using two experimental media with different chemical components. For experimentation, the measure of the transepithelial electrical resistance (TEER) allowed us to evaluate the state of cells; and the quantities of bepridil have been quantified using a gas chromatography/mass spectrometry system. First, when using the medium alone, without bepridil, Caco-2 cell integrity is, at least, maintained for 8 h using both media. However, for 24-h studies, only the DMEMc medium, rich in essential nutriments, allowed cell integrity to be maintained. Then, with bepridil in HBSS medium, the TEER measurement showed a dose-dependent toxic effect of bepridil, whereas in the DMEMc medium, the toxic effect was only found for the highest dose (12 microg). This difference is probably related to the high binding of bepridil to proteins of the DMEMc medium, therefore minimising the concentration of the free compound. The kinetics of bepridil result from two phenomena: first, an immediate passage of a slight part of bepridil through the cell barrier and second, a high retention of most of the bepridil dose in the cell level. The transfer of bepridil from the apical to the basolateral compartment appears quantitatively and kinetically different using DMEMc or HBSS medium. The retention of the compound in the 'filter with Caco-2 cells' compartment is higher in DMEMc medium (60% at 3 microg) than in HBSS medium (46% at 3 microg), and bepridil entering the basolateral compartment is delayed in the DMEMc medium. This study exhibits the importance of the selected medium on results and interpretation of data and the predominance of DMEMc to study the transport of lipophilic compounds highly retained in cells. PMID- 10370217 TI - Are calculated log P values for some guanine derivatives by different computer programs reliable? AB - The log P values of n-octanol/water for some guanine derivatives, acyclovir, deoxyaciclovir and their acetyl congeners, were calculated by some commercially available computer programs for log P calculation. These values were compared with those obtained by the conventional shake-flask method. It was established that the calculations of log P values for examined guanine derivatives by these computation programs do not give reliable results. PMID- 10370218 TI - Thermally reversible xyloglucan gels as vehicles for oral drug delivery. AB - The potential, as sustained release vehicles, of gels formed in situ following the oral administration of dilute aqueous solutions of a xyloglucan polysaccharide derived from tamarind seed has been assessed by in vitro and in vivo studies. Aqueous solutions of xyloglucan that had been partially degraded by beta-galactosidase to eliminate 44% of galactose residues formed rigid gels at concentrations of 1.0 and 1.5% w/w at 37 degrees C. The in vitro release of indomethacin and diltiazem from the enzyme-degraded xyloglucan gels followed root time kinetics over a period of 5 h at 37 degrees C at pH 6.8. Plasma concentrations of indomethacin and diltiazem, after oral administration to rats of chilled 1% w/w aqueous solutions of the enzyme-degraded xyloglucan containing dissolved drug, and a suspension of indomethacin of the same concentration were compared. Constant indomethacin plasma concentrations were noted from both formulations after 2 h and were maintained over a period of at least 7 h. Bioavailability of indomethacin from xyloglucan gels formed in situ was increased approximately threefold compared with that from the suspension. The results of this study suggest the potential of the enzyme-degraded xyloglucan gels as vehicles for oral delivery of drugs. PMID- 10370219 TI - Clonazepam release from bioerodible hydrogels based on semi-interpenetrating polymer networks composed of poly(epsilon-caprolactone) and poly(ethylene glycol) macromer. AB - Poly(ethylene glycol)(PEG) macromers terminated with acrylate groups and semi interpenetrating polymer networks (SIPNs) composed of poly(epsilon caprolactone)(PCL) and PEG macromer were synthesized to obtain a bioerodible hydrogel. Polymerization of PEG macromer resulted in the formation of cross linked gels due to the multifunctionality of macromer. Glass transition temperature (Tg) and melting temperature (Tm) of PEG networks and PCL in the SIPNs were inner-shifted, indicating an interpenetration of PCL and PEG chains. Water content in the SIPNs increased with increasing PEG weight fraction due to the hydrophilicity of PEG. The amount of clonazepam (CNZ) released from the SIPNs increased with higher content in the SIPNs, lower drug loading, lower concentration of PEG macromer during the SIPNs preparation, and higher molecular weight of PEG. In particular, a combination with low PEG content and low CNZ solubility in water led to long-term constant release from these matrices in vitro and in vivo. PMID- 10370220 TI - Design of a fluid energy single vessel powder processor for pharmaceutical use. AB - This study introduces a motionless novel single vessel powder processor designed to carry out all of the unit operations in the preparation of powders for tableting. The processor used controllable fluid dynamics to provide the energy for each unit operation. The vessel design was evaluated using a computational fluid dynamics model which indicated the flow necessary for the intended processing operations to take place. The processor performance was evaluated experimentally for two unit processes: particle size reduction and dry powder mixing. The processor was found capable of reducing the size of lactose granules from a median particle diameter of 459 microm to a median particle diameter of 182 microm within 5 min under optimal process conditions. It was found that a formulation containing lactose granules (373 microm median particle diameter) and a model drug, sodium chloride (30 microm), could be mixed to an improved degree of homogeneity in comparison with equivalent powders blended using a conventional turbulent tumbling technique. It was concluded that a processor having controllable fluid dynamics offered the potential to perform multi-task processing of powders. PMID- 10370221 TI - Effect of ethanol/propylene glycol on the in vitro percutaneous absorption of aspirin, biophysical changes and macroscopic barrier properties of the skin. AB - The effect of the solvent systems ethanol (EtOH), propylene glycol (PG) and combinations thereof was examined on the in vitro percutaneous absorption of the antithrombotic, aspirin, through porcine epidermis. Biophysical changes in the stratum corneum lipids were studied through the use of Fourier transform infrared (FTIR) spectroscopy. Macroscopic barrier properties of the epidermis were examined through the use of in vitro transepidermal water loss (TEWL). The flux of aspirin increased with increasing concentrations of EtOH in the solvent systems. The maximum flux of aspirin was achieved by 80% EtOH in combination with 20% PG beyond which (i.e. 100% EtOH) there was no increase in the flux. FTIR spectroscopic study was enacted in order to determine the biophysical properties of the stratum corneum when the solvents were applied. The FTIR spectra of the stratum corneum treated with 80% EtOH/20% PG showed a maximum decrease in absorbance for the asymmetric and symmetric C&z. sbnd;H peaks, which suggests a greater loss of the lipids in the stratum corneum layers. In vitro TEWL studies allowed an investigation into the macroscopic barrier integrity properties of the stratum corneum. The TEWL results indicated that each of the solvent systems significantly enhanced (P<0.05) in vitro TEWL in comparison to the control. In conclusion, 80% EtOH/20% PG enhanced the percutaneous absorption of aspirin by perturbing the macroscopic barrier integrity of the stratum corneum and through a loss of stratum corneum lipids. PMID- 10370223 TI - Cytomegalovirus retinitis revealing a leukemic gamma/delta T-cell lymphoma. PMID- 10370224 TI - Gonadal and Extragonadal Functions of Ad4BP/SF-1: Developmental Aspects. AB - Gene disruption studies have clearly indicated that 4-binding protein/steroidogenic factor 1 (Ad4BP/SF-1), originally identified as a steroidogenic tissue-specific transcription factor, plays crucial roles in the process of non-steroidogenic as well as steroidogenic tissue differentiation. Although the precise mechanisms underlying differentiation of these tissues are still under investigation, spatial and temporal expression profiles of the gene encoding Ad4BP/SF-1 support its contribution to tissue development from the earliest stages of ontogeny. PMID- 10370225 TI - Corticotropin-releasing Hormone in Human Pregnancy and Parturition. AB - The role of corticotropin-releasing hormone (CRH) in the regulation of pituitary adrenocorticotropin secretion and the stress response is well established. However, in recent years this peptide has been found to serve a number of functions outside the classic neuroendocrine domain. During pregnancy, CRH derived from the placenta is thought to play a crucial role in the regulation of foetal maturation and the timing of delivery, and CRH has also been implicated in the control of foetal-placental bloodflow. Abnormalities of the placental CRH system might be involved in the pathogenesis of preterm labour, foetal growth retardation and pre-eclampsia, which are the three leading causes of perinatal morbidity and mortality in developed countries. PMID- 10370226 TI - Pregestational Diabetes Mellitus and Pregnancy. AB - In a pregnancy complicated by diabetes the foetus is at increased risk of congenital malformations, macrosomia and stillbirth. For the mother, diabetes complications (retinopathy nephropathy) may worsen, and in this group there is a higher incidence of hypertension, pre-eclamptic toxaemia and intrauterine growth restriction. However, women with good control who are free of diabetic complications can expect a normal outcome. PMID- 10370228 TI - Regulation of the FSH Receptor in the Ovary. AB - The follicle-stimulating hormone receptor (FSHR) is found exclusively on granulosa cells from as early as the two-layer or primary stage of folliculogenesis. Up to four alternatively spliced transcripts have been described. An increase in steady-state levels of FSHR mRNA as well as a change in alternative splicing appear to be important during early folliculogenesis. FSHRs remain on the granulosa cells of healthy follicles until they become atretic or luteinize, with relatively little change in receptor number per cell. Factors that increase expression are FSH itself, cAMP and its analogs, activin and transforming growth factor beta (TGF-beta), whereas epidermal growth factor (EGF)/TGF-alpha decrease expression. The regulatory regions of the FSHR gene are more akin to 'housekeeping' genes than to a highly regulated gene, and there is an E box in the promoter region. Regulation of the initial expression of FSHR mRNA at the two-granulosa cell layer stage of folliculogenesis is not understood. It will also be important to understand whether the alternatively spliced FSHR transcripts and their prevalence in the ovary during early folliculogenesis represent a biologically important phenomenon. PMID- 10370229 TI - Regulation of the Primate Corpus Luteum: Cellular and Molecular Perspectives. AB - The process of luteinization, during which follicular granulosa cells are transformed into luteal cells, is accompanied by dramatic changes in the responses of luteal cells to luteinizing hormone (LH) and cAMP. The goal of this review is to summarize the findings of recent studies in monkeys, which have shown that the expression of the cAMP-dependent nuclear transcription factor CREB (cAMP-response element-binding protein) ceases upon luteinization, and also to suggest possible consequences of its loss on corpus luteum function and lifespan. PMID- 10370230 TI - Aging and Muscle Loss. AB - Aging is associated with a decrease in fat-free mass, an increase in fat mass, and progressive impairment of muscle function and performance. Diminishing anabolic hormone levels and progressive declines in muscle protein turnover contribute to the multifactorial pathophysiology of age-associated sarcopenia. The potential effects of anabolic hormone replacement on body composition and functional capacity are only beginning to be studied. PMID- 10370231 TI - A guide to the WWW PMID- 10370232 TI - Quality control and assurance from the development to the production of biopharmaceuticals. AB - Consumer and patient safety have become the prerequisites for (bio)pharmaceutical product development, production and marketing. The ability to provide an effective, pure, safe product is the primary factor determining the product's success. However, with an ever-increasing number of national and international regulations, 'quality assurance' has acquired a threatening ring for many project managers. Many think that ensuring and improving quality is expensive, but regulations aid public acceptance. Good manufacturing practice can be developed into a business asset and need not be seen as merely a regulatory hurdle. PMID- 10370233 TI - Biotechnology as a route to nanotechnology AB - In manufacturing, it is common to hold, position and assemble parts in a fashion that is not possible today at the molecular scale, yet the basic principles of positional assembly are just as applicable whether the parts are meters or nanometers in size. The ability to hold and position parts gives us remarkable flexibility, whether we are making furniture or synthesizing complex molecular structures. Applying this powerful concept at the molecular scale will require the development of new tools and pose new challenges in many fields, but the rewards will be enormous. PMID- 10370234 TI - Integrated pathway-genome databases and their role in drug discovery. AB - Integrated pathway-genome databases describe the genes and genome of an organism, as well as its predicted pathways, reactions, enzymes and metabolites. In conjunction with visualization and analysis software, these databases provide a framework for improved understanding of microbial physiology and for antimicrobial drug discovery. We describe pathway-based analyses of the genomes of a number of medically relevant microorganisms and a novel software tool that visualizes gene-expression data on a diagram showing the whole metabolic network of the microorganism. PMID- 10370235 TI - Basidiomycetes as new biotechnological tools to generate natural aromatic flavours for the food industry AB - Consumer preference for natural food additives has led to an increasing demand for natural aromatic compounds. An alternative production process to plant and chemical sources is the use of biotechnological methods involving microorganisms, which ensure a stable supply, quality and price. Among filamentous fungi, white rot basidiomycetes represent an important group that generate a wide range of flavouring compounds, particularly aromatic molecules. Their biotechnological potential to produce natural aromatic flavours de novo or by biotransformation thus presents a very interesting challenge. PMID- 10370237 TI - Optical mapping and its potential for large-scale sequencing projects. AB - Physical mapping has been rediscovered as an important component of large-scale sequencing projects. Restriction maps provide landmark sequences at defined intervals, and high-resolution restriction maps can be assembled from ensembles of single molecules by optical means. Such optical maps can be constructed from both large-insert clones and genomic DNA, and are used as a scaffold for accurately aligning sequence contigs generated by shotgun sequencing. PMID- 10370236 TI - Exploiting antibody-based technologies to manage environmental pollution. AB - Many commodities used in the food, cosmetic, chemical and environmental industrial sectors function by binding to other molecules. Antibodies can bind virtually any molecule, from large proteins to small organic ligands, and could replace substances with undesirable medical, social or environmental side effects, if they could be provided in stable configurations and in quantities and at costs acceptable to industry. Antibody fragments also offer the possibility of sensitive detection and efficient removal of organic pollutants from the environment. PMID- 10370238 TI - First the CDKs, now the DDKs. AB - In budding yeast, Dbf4p and Cdc7p control initiation of DNA synthesis. They form a protein kinase - Cdc7p being the catalytic subunit and Dbf4p a cyclin-like molecule that activates the kinase in late G1 phase. Dbf4p also targets Cdc7p to origins of replication, where probable substrates include certain Mcm proteins. Recent studies have identified Dbf4p- and Cdc7p-related proteins in fission yeast and metazoans. These homologues also phosphorylate Mcm proteins and could have a similar function to that of Dbf4p-Cdc7p in budding yeast. Thus, it seems likely that, like the cyclin-dependent kinases (CDKs), the Dbf4p-Cdc7p activity is conserved in all eukaryotes. PMID- 10370239 TI - Is dynamin a regular motor or a master regulator? PMID- 10370240 TI - Germline cysts: a conserved phase of germ cell development? AB - Germ cells in many vertebrate and invertebrate species initiate gametogenesis by forming groups of interconnected cells known as germline cysts. Recent studies using Xenopus, mouse and Drosophila are beginning to uncover the cellular and molecular mechanisms that control germline cyst formation and, in conjunction with morphological evidence, suggest that the process is highly conserved during evolution. This article discusses these recent findings and argues that cysts play an important and general role in germ line development. PMID- 10370241 TI - Hsp90's secrets unfold: new insights from structural and functional studies. AB - Hsp90 is a molecular chaperone associated with the folding of signal-transducing proteins, such as steroid hormone receptors and protein kinases. Results from recent studies have shed light on the structure of Hsp90 and have demonstrated that it can bind to and hydrolyse ATP. Hsp90 forms several discrete subcomplexes, each containing distinct groups of co-chaperones that function in folding pathways. Although Hsp90 is not generally involved in the folding of nascent polypeptide chains, there is a growing list of proteins whose activity depends on its function, including heat-shock factor. This review addresses recent developments in our understanding of the structure and function of Hsp90. PMID- 10370242 TI - Occludin and claudins in tight-junction strands: leading or supporting players? AB - Tight junctions have attracted much interest from cell biologists, especially electron microscopists, since on freeze-fracture electron microscopy they appear as a well-developed network of continuous, anastomosing intramembranous strands (tight-junction strands). These strands might be directly involved in the 'barrier' as well as 'fence' functions in epithelial and endothelial cell sheets, but until recently little was known of their constituents. This review discusses current understanding of the molecular architecture of tight-junction strands, focusing on the recent discovery of two distinct types of tight-junction-specific integral membrane proteins, occludin and claudins. PMID- 10370243 TI - Regulation of Smad signalling by protein associations and signalling crosstalk. AB - Smads are intracellular signalling mediators for the family of transforming growth factor beta (TGF-beta)-related growth and differentiation factors, which signal through transmembrane serine/threonine kinases. Following receptor-induced activation, heteromeric Smad complexes translocate into the nucleus, where they act as transcription factors. Recent progress has revealed that Smad signalling is not merely determined by activation of the class of TGF-beta receptors, but is also regulated through crosstalk with other kinase signalling cascades. In addition, the Smads regulate transcription through functional cooperativity and physical interactions with other transcription factors, which might also be targets for regulation by other signalling cascades. This signalling crosstalk might explain the complexity of the responses to TGF-beta and related factors. PMID- 10370244 TI - Richard W. Young: and the band marched on. PMID- 10370245 TI - Use of caged fluorochromes to track macromolecular movement in living cells. AB - One way to visualize and track the movement of macromolecules in the living cell is to follow their movement after tagging the molecule with a 'caged' or chemically masked fluorochrome. The fluorochrome does not fluoresce until the caging group is released by spot photoactivation, and the bright fluorescent signal can then be tracked as it moves into the dark surrounding area of the cell. When coupled with rapid imaging microscopy, it is possible to measure rates of movement as fast as macromolecular diffusion. This article describes the use of photoactivatable fluorochromes to track the intracellular movement of both proteins and nucleic acids and to track cell lineages. PMID- 10370246 TI - Careers-perspective interview. PMID- 10370247 TI - ERM proteins in cell adhesion and membrane dynamics: Authors' correction PMID- 10370248 TI - Evidence for multi-functional interactions in early visual motion processing. PMID- 10370249 TI - Cytoskeletal dynamics in dendritic spines: direct modulation by glutamate receptors? AB - A wide heterogeneity in dendritic-spine morphology is observed and ultrastructural changes can be induced following experimental stimulation of neurons. Morphological adaptation of a given spine might, thus, reflect its history or the current state of synaptic activity. These changes could conceivably result from rearrangements of the cytoskeleton that is subjacent to excitatory synapses. This article dicusses the direct and indirect interactions, between glutamate receptors and the cytoskeletal proteins, which include PDZ containing proteins, actin and tubulin, as well as associated proteins. In fact, the synaptic-activity-controlled balancing of monomeric, dimeric and polymeric forms of actin and tubulin might underlie the changes in spine shape. These continuous adaptations could be relevant for physiological events, such as learning and the formation of memory. PMID- 10370250 TI - Innate and adaptive immune responses can be beneficial for CNS repair. AB - The limitation of immune responsiveness in the mammalian CNS has been attributed to the intricate nature of neuronal networks, which would appear to be more susceptible than other tissues to the threat of permanent disorganization when exposed to massive inflammation. This line of logic led to the conclusion that all forms of CNS inflammation would do more harm than good and, hence, the less immune intervention the better. However, mounting evidence indicates that some forms of immune-system intervention can help to protect or restore CNS integrity. We have shown that the innate immune system, represented by activated macrophages, can facilitate the processes of regeneration in the severed spinal cord. More recently, we found that autoimmune T cells that are specific for a component of myelin can protect CNS neurons from the catastrophic secondary degeneration, which extends traumatic lesions to adjacent CNS areas that did not suffer direct damage. The challenge, therefore, is to learn how to modify immune interactions in the traumatized CNS in order to promote its post-injury maintenance and repair. PMID- 10370251 TI - Not only how, but also what and why. PMID- 10370252 TI - Reply. PMID- 10370253 TI - Corrigendum. PMID- 10370254 TI - Gauging sensory representations in the brain. AB - The stream of information that enters a sensory system is a product of the ecological niche of an organism and the way in which the information is sampled. The most salient characteristic of this sensory stream is the rich temporal structure that is caused by changes in the environment and self motion of sensors (for example, rapid eye or whisker movements). In recent years, substantial progress has been made in understanding how such rapidly varying stimuli are represented in the responses of sensory neurons of a large variety of sensory systems. The crucial observation that has emerged from these studies is that individual action potentials convey substantial amounts of information, which permits the discrimination of rapidly varying stimuli with high temporal precision. PMID- 10370255 TI - The retrosplenial cortex and emotion: new insights from functional neuroimaging of the human brain. AB - Little is known about the function of the retrosplenial cortex and until recently, there was no evidence that it had any involvement in emotional processes. Surprisingly, recent functional neuroimaging studies show that the retrosplenial cortex is consistently activated by emotionally salient words. A review of the functional neuroimaging literature reveals a previously overlooked pattern of observations: the retrosplenial cortex is the cortical region most consistently activated by emotionally salient stimuli. Evidence that this region is also involved in episodic memory suggests that it might have a role in the interaction between emotion and episodic memory. Recognition that the retrosplenial cortex has a prominent role in the processing of emotionally salient stimuli invites further studies to define its specific functions and its interactions with other emotion-related brain regions. PMID- 10370256 TI - CART peptides: novel addiction- and feeding-related neuropeptides. AB - CART peptides are novel, putative brain-gut neurotransmitters and co-transmitters that probably have a role in drug abuse, the control of feeding behavior, sensory processing, stress and development. They are abundant, processed and apparently released. Exogenously applied peptides cause inhibition of feeding and have neurotrophic properties. Although the precise sequences, relative abundance and efficacy of all CART peptides are currently being determined, small molecules that are active at putative CART receptors could have substantial therapeutic promise. PMID- 10370258 TI - Sperm displacement in yellow dung flies: a role for females. PMID- 10370259 TI - Statistical analysis of microsatellite DNA data. PMID- 10370257 TI - Patterning motoneurons in the vertebrate nervous system. AB - Vertebrate motoneurons show considerable diversity in their soma locations, axonal trajectories and innervation targets. Results from studies of a variety of vertebrate species as well as fruit-flies are elucidating the mechanisms by which this diversity is generated. Motoneuron subpopulations appear to be defined by combinations of transcription factor genes expressed in distinct spatiotemporal patterns in both motoneuron progenitors and postmitotic motoneurons. Notochord derived signals can induce motoneuron formation, paraxial-mesoderm-derived signals can pattern motoneuron subpopulations along the rostrocaudal body axis, and local signals within the neural tube can regulate the number and time at which motoneurons form. Additional, later signals can promote formation of proper central circuitry and motoneuron survival. The identification of the genes and signals responsible for regulating these processes should help to provide a more detailed understanding of motoneuron patterning. PMID- 10370261 TI - Pterosaurs: back to the traditional model? AB - For most of the past 200 years, pterosaurs have been reconstructed with wing membranes attached to both the forelimbs and hindlimbs and with a quadrupedal stance and gait when grounded. In the early 1980s, this traditional model was replaced by a new design with narrow, stiff wings confined to the forelimbs and an upright, bipedal, digitigrade stance and gait. However, new studies of complete, uncrushed skeletal remains, well preserved wing membranes and extensive new pterosaur tracks, combined with reanalysis of the relationships of pterosaurs to other reptiles, suggest that a return to the traditional model is now overdue. PMID- 10370260 TI - Fatal fighting in fig wasps - GBH in time and space. PMID- 10370262 TI - Relating populations to habitats using resource selection functions. AB - Habitat use can be characterized by resource selection functions (RSFs) that are proportional to the probability of an area being used by an animal. We highlight two procedures that have recently been used to relate RSFs to population density, dependent upon which field procedures are practical for a species. These new developments allow RSF models to be interfaced with geographical information systems (GIS) to map the probability of use, and ultimately populations, across landscapes. PMID- 10370263 TI - Patch structure, dynamics and implications for the functioning of arid ecosystems. AB - Arid ecosystems present a two-phase mosaic structure of high- and low-cover patches. Vegetation patches differ among ecosystems in size and shape. However, recent studies indicate striking similarities in patch dynamics and in mechanisms explaining their origin and maintenance. Two major types of system, banded and spotted vegetation, which are characterized by patch shape, both originate from common mechanisms, although each is dominated by a different driver. Banded vegetation occurs when water is the dominant driver of the redistribution of materials and propagules, whereas spotted vegetation results when wind is the major redistribution driver. Model analysis indicates that patchy vegetation structure enhances primary production. PMID- 10370265 TI - Extinction by infection. PMID- 10370264 TI - Reply from L. Bromham, D. Penny and M.J. Phillips. PMID- 10370266 TI - Coastal marine communities: trends and perspectives from human-exclusion experiments. AB - The ecological roles of humans in marine communities have been poorly studied. Humans have special characteristics, such as culture, and are perceived as complex ecological actors. Observations and experiments conducted in coastal (rocky intertidal and nearshore) 'no-take' areas or reserves in Chile and around the world have permitted a better understanding of the role played by humans as top predators and the resulting trophic-cascade effects along the food-webs. These studies have revealed an urgent need to incorporate humans into ecological studies and have helped to promote links between ecology and social sciences. PMID- 10370268 TI - Linking biodiversity information sources. PMID- 10370267 TI - Causes and consequences of recent freshwater invasions by saltwater animals. AB - Transitions from marine to freshwater habitats constitute dramatic shifts between 'adaptive zones' that have initiated the radiation and speciation of many taxa. As recently as 10?000 years ago, deglaciation resulted in marine fauna being trapped in freshwater lakes. In modern times, human activity has caused the acceleration of freshwater invasions from marine or brackish habitats, leading to serious environmental problems. The rapid pace of these invasions provides ideal opportunities for examining initial responses to environmental change and mechanisms involved in habitat transitions. Despite conservation implications and evolutionary applications, recent transitions to fresh water remain inadequately explored. PMID- 10370269 TI - Tapping artificially into natural talents. PMID- 10370271 TI - Phase locking: another cause of synchronicity in predator-prey systems. PMID- 10370270 TI - Molecular dates and the mammalian radiation. PMID- 10370272 TI - DOs' approach to patient is 'wholistic'--not holistic. PMID- 10370273 TI - Screening for hemochromatosis can prevent serious disease. PMID- 10370274 TI - Asthma: an underappreciated disease with a huge impact on society. PMID- 10370275 TI - Updated HIV guidelines available. PMID- 10370276 TI - Beyond the 'box'. PMID- 10370277 TI - Domestic violence: effect on pregnancy outcome. AB - A predominantly white, suburban, indigent population of pregnancy women were followed up to determine the incidence of domestic violence and its effect on preterm delivery, low birth weight, and outcome of pregnancy (infant admission to the neonatal intensive care unit. A total of 489 gravidas were screened for domestic violence and drug and alcohol abuse. Patients were assigned to the control group if they had no substance abuse and no domestic violence and to the study group if they had no substance abuse but were victims of domestic violence. Of the total study population, 20% were victims of domestic violence. Among patients suffering domestic violence, 22% had preterm deliveries as compared with 9% of patients without domestic violence (P = .002). Sixteen percent of patients in the study group had low-birth-weight babies compared with 6% of women in the control group (P = .002). No significant relationship was found between domestic violence and admission to the neonatal intensive care unit. Therefore, domestic violence is a risk factor for preterm delivery and low-birth-weight infants. PMID- 10370278 TI - An osteopathic approach to asthma. AB - Asthma has become a serious challenge to clinical medicine today, with an increase in incidence, morbidity, and mortality over the past two decades. Asthma continues to be a problem despite increased knowledge of the pathophysiology of asthma coupled with the development of a variety of new and innovative medications that can be used to treat asthma. Five areas involving asthma management are reviewed and involve a failure to do the following: (1) identify disease instability and progression; (2) adopt an optimal pharmacologic treatment plan; (3) identify and help the patient avoid environmental triggers; (4) evaluate and treat certain disruptive psychodynamic issues; and (5) use essential non-pharmacologic modes of therapy such as osteopathic manipulation, nutritional considerations, physical training, and controlled breathing techniques that may help to favorably modify the asthma disease process. PMID- 10370279 TI - Quality medical management of the geriatric population using practice guidelines, physician-managed home health services, and continuous quality-improvement management strategies. AB - One challenge confronting physicians in the 1990s is the delivery of cost effective quality healthcare to the elderly. Given current utilization, federal expenditures on Medicare and other federal healthcare programs are projected to increase to more than 6% of the gross national product by the year 2030. Physicians' practice habits will have an impact on the cost of caring for the nation's elderly. It is therefore essential for physicians to recognize their role in delivering quality geriatric care. First, they must be active and diligent in the pursuit of practice guidelines that support changes in the current standards of care. Second, physicians should consider alternative approaches to present practice patterns such as physician-managed home health services. Finally, physicians need to take a proactive approach to quality by supporting continuous quality improvement. The purpose of this review is to present some of the existing established practice guidelines, data to support the use of physician-managed home care as an alternative practice approach, and suggestions to incorporate the principles of continuous quality improvement to serve as one practice model that can be used by physicians to improve the quality of medical care given to America's elderly. PMID- 10370280 TI - Pancreatic cystic neoplasms. AB - Cystic neoplasms of the pancreas are relatively rare. This makes the evaluation and treatment of these tumors widely varied. The authors describe a patient who came to our hospital with complaints of abdominal pain, but no other related symptoms. Diagnostic evaluation of the patient yielded normal results, except for inspection and palpation of the abdominal areas, which revealed a large epigastric mass; this finding was confirmed subsequently by ultrasonographic examination and computed tomographic scanning. This article presents the case and reviews the literature, specifically related to diagnosis and current treatments. PMID- 10370281 TI - Scripted role play: a technique for teaching sexual history taking. AB - Many inexperienced physicians are uncertain as to how to go about gathering a sexual history. They have had courses in basic history taking, but they do not know how to go about obtaining the information in a nonthreatening way. Their opportunities for interaction with real patients have typically been limited. Scripted role play is a very effective method for providing residents with an opportunity to enhance their sexual-history-taking skills. This article describes scripted role play as a teaching method; explains why it is effective for teaching sexual history taking; presents general guidelines for script writing for this purpose; details how to organize a teaching session; and provides some tips for the session facilitator. PMID- 10370282 TI - [The meaning of duodenostasis in the intestinal phase gastric secretion disorders in bleeding duodenal ulcer]. AB - The data on the duodenal motor-evacuational function investigation in 62 patients with an ulcer bleeding are presented. It was established that the degree of duodenal chronic impassability is correlating with the degree of disorders of the gastric secretion intestinal phase. That's why it is necessary to stipulate the elimination of hyperchlorhydria in all phases of gastric secretion while such complication correction. PMID- 10370284 TI - [The influence of Helicobacter pylori on the surgical outcome of duodenal ulcer]. AB - There were examined 79 patients, operated on for duodenal ulcer disease. Before the operation the Helicobacter pylori (HP) dissemination of stomach and duodenum of significant degree was diagnosed in 51.9%, mild degree--in 34.2% and of a nonsignificant one--in 13.9% of patients. After the operation the HP dissemination of stomach and duodenum had persisted. PMID- 10370283 TI - [Clinical-endoscopic factors of gastro-duodenal ulcer associated with Helicobacter pylori in elderly and senile patients]. AB - Clinical peculiarities of the disease were detected, basing on the analysis of observation of 73 elderly and senile patients with gastroduodenal ulcer disease. While the gastric ulcer disease presence Helicobacter pylori was revealed in 71.4% of patients, aged 60-74 years old, and in 63.3%--aged older than 75 years, while duodenal ulcer disease--in 83.8 and 70% accordingly. PMID- 10370285 TI - [The role of a free radical mechanisms in intestinal obstruction during peritonitis and its repair]. AB - In 53 patients with diffuse peritonitis in a toxical phase there was investigated the dynamics of the gut functional impassability regress after the operation. There were adduced the grounds for application of the preparations with antihypoxical and antioxidant properties in complex therapy of such patients. The method of selective spectral peripheric electrogastroenterography was introduced as a noninvasive method of computeric monitoring of the gut functional state. PMID- 10370286 TI - [Blood circulation in the liver after distal spleno-renal anastomosis in cirrhosis]. AB - Complex hemodynamical investigations were performed in 24 patients with liver cirrhosis before and after the distal splenorenal anastomosis (DSRA) conduction according to conventional and modified methods. After the conventional DSRA formation the portal blood flow had reduced down by 39.6% and after modified method--by 14%. Linear speed of the blood flow in a portal vein after the conventional DSRA formation had reduced a twice, after the modified DSRA--did not change. The pressure in a portal vein after the conventional DSRA formation had reduced by 35%, and after the modified DSRA--by 11%. The portal perfusion lowering had promoted the compensational increase of the blood flow in hepatic artery after the conventional DSRA forming by 22% and after the modified DSRA--by 8%. PMID- 10370287 TI - [Causes of unsatisfactory outcome in patients with diabetes mellitus complicated by necrotic inflammation of the foot]. AB - Retrospective analysis of the conservative and surgical treatment results of 222 patients with diabetes mellitus, complicated by necrotic-inflammatory affection of foot, was performed for the 1991-1996 period. In 25.2% of observations the extremity amputation was done on the hip level, 13 (5.8%) of patients died. The unsatisfactory result causes were the reoperation performance on foot in a critical ischemia zone, the decrease of the first operation volume, nonadequate correction of metabolic, haemorheologic disturbances, antibacterial therapy. PMID- 10370288 TI - [Reinnervation of a skeletal muscle after its directed injury]. AB - The connection of the motor nerve stump with a muscle permits to achieve the definite standard of the muscle structures reinnervation, but this process is limited the nerve localization zone. Morphological substantiation of the muscle reinnervation method in experiment is proposed, including the fascicules preparation and accommodation of the nerve-neurotizer in a muscle depth, the neurotropismus stimulation using the directed damage of the muscle tissue. The method was tested in clinic in 9 patients. PMID- 10370290 TI - [Chylothorax]. AB - Six cases of chylothorax of their own observation were depicted. In one of them the cause of the disease was not disclosed. In four patients a pleural puncture or pleurodesis were accomplished, in two--an operation was performed. PMID- 10370289 TI - [Hormonal, immune and metabolic status in intraoperative injury in the course of electrical stimulation with propofol]. AB - There are presented the results of complex examination of neurohumoral and immune systems in patients, operated in conditions of electroimpact with propofol. Were determined the significant reduction of quantity and redistribution of activated lymphocytes, the rise of secondary immunodeficiency state while preservation of stable indexes of neuroendocrine system. PMID- 10370291 TI - [Clinical course and surgical treatment of complex forms of recurrent acute paraproctitis]. AB - Clinical course and results of surgical treatment of 94 patients with complex forms of recurrent acute paraproctitis were analysed. Radical surgical intervention was conducted in 68 patients, of whom in 35--using the pararectal abscess opening with cryptectomy, in 17--with translocation of a distal rectum mucosa, in 16--with a ligature conduction through internal aperture of fistula. One-nine year follow-up of 89 patients is available. After radical intervention the recurrence and postoperative complications had occurred in 19%, after nonradical one--in all the patients. PMID- 10370292 TI - [Pathogenesis, surgical treatment and prophylaxis of goiter recurrence]. AB - The causes of the nontoxic goiter recurrency occurrence, the prophylactic methods and the results were analysed according to the data obtained while examination and treatment of 115 patients. The autoimmune damage of a leaved thyroidal gland remnant constitutes one of the causes of the genuine recurrency of goiter. PMID- 10370293 TI - [Minimally invasive therapy of the cystic-transformed thyroidal nodes]. AB - The possibility of the miniinvasive methods application in the treatment of a cystic-transformed nodes of thyroid gland was studied up. In 59 patients the diagnostical and therapeutical interventions were carried out: in 39--the node's liquid contents aspiration (in 2 patients while cytological investigation the malignant transformation was revealed with operative treatment recommended); in 19--the ethanol sclerosing therapy (in 14--single, in 5--two-stage according to original method); in 4--the laser photocoagulation. The above mentioned methods efficacy was 82.2, 92.8 100 and 100% accordingly. PMID- 10370294 TI - [The influence of the efferent and quantum methods of treatment on endotoxemia pathogenetical factors, the course and outcome of burns]. AB - There were observed 424 injured persons with severe and extremely severe burn and endotoxemia of a high degree. In 176 of them (the main group) for the purpose of additional nonspecific detoxication were applied an efferent and quantum methods: plasmapheresis, indirect electrochemical oxidation of blood, the laser and ultraviolet irradiation of blood. In the main group 23.3% of patients died and in a control one--62.5%. It was noted the reduce of the most severe complication of the burn disease--sepsis (an early one--in 5 times and a late one--in 1.5 times). The survival index in the main group while the sepsis had appeared was three times higher than in a control one. PMID- 10370295 TI - [The application of endolymphatic polychemotherapy in inoperative cancer of the lungs]. AB - In 164 patients with inoperable pulmonary cancer endolymphatical infusion of cyclophosphamide and metotrexate had promoted the efficacy and security promotion of polychemotherapy conduction in comparison with such a therapy, using intravenous introduction of preparations. It was noted the increase of a life span and a five-year survival of the patients. PMID- 10370296 TI - [First experiment in the application of fibrin glue "Bioadhesive" in the surgical treatment of mammary gland tumors]. AB - The method of prophylaxis and treatment of an early complications of surgical treatment of the mammarial gland tumors using native biological fibrinous glue "Bioadhesive" was proposed. The preparation application had permitted to reduce the operative wound healing duration, to reduce a wound exudate quantity, the duration and intensity of lymphorrhea and the seroma rise frequency, and to improve significantly an operative intervention cosmetic result. PMID- 10370297 TI - [Methods of treatment of the mammary gland cancer in its initial stage]. AB - The results of treatment of 1231 women patients with the mammarial gland cancer of initial stages was analysed, in 151 of them the organ-preserving operation was performed. The results of treatment with five-year follow-up, using the organ preserving operations do not cede to such in application of classical mastectomy. PMID- 10370298 TI - [Experimental models of abdominal aorta aneurysms]. PMID- 10370299 TI - [Statistical methods used for the evaluation of disease severity and surgical outcome]. PMID- 10370300 TI - [Giant cyst of the left lung complicated by mediastinal hernia and hemoptysis]. PMID- 10370301 TI - [The treatment of the extra-adrenal pheochromocytoma]. PMID- 10370302 TI - [Amino acid composition of the bile in gallbladder inflammatory diseases]. PMID- 10370303 TI - [Observation of giant Zenker's diverticulum complicated by esophageal obstruction]. PMID- 10370304 TI - [The application of endoscopic methods in the treatment of vesico-ureteral reflux in children with the use of polyacrylamide hydrogel "Interfall"]. PMID- 10370305 TI - [Regional scientific-practical conference of surgeons on "Current issues of emergency surgical services for abdominal organs"]. PMID- 10370306 TI - [Classification and selection of treatment of chronic gastroduodenal ulcer complicated by hemorrhage in elderly and senile patients]. AB - The classification of chronical bleeding gastroduodenal ulcer in elderly and senile patients, considering the blood loss severity and concurrent pathology, was proposed. The tactics of treatment--conservative, operative, or actively waiting one--was determined according to the classification adduced. The proposed classification application and the tactics of treatment also have permitted to reduce mortality from 15.7 to 11.2%. PMID- 10370307 TI - [Purulent pancreatitis]. AB - For last 12 years 455 patients with an acute pancreatitis were treated in the clinic. In 29 (6.3%) of them purulent pancreatitis, originating from pancreonecrosis, was diagnosed. Operative intervention consisted of laparotomy conduction, bursa omentalis draining through lig, gastrocolicum, excision of purulent-necrotic foci in pancreas and retroperitoneal fat, draining purulent cavity using two silicon tubes, introduced via left and right epigastral regions, durable lavage of purulent cavity with antiseptic solutions. Five patients died. PMID- 10370308 TI - [Surgical treatment of an acute pancreatitis]. AB - The results of surgical treatment of 107 patients with an acute pancreatitis for 1990-1998 period were analysed. The miniinvasive methods application, directed on the aseptic pancreatic necrosis transition to purulent pancreatitis prophylaxis and timely conduction of operative intervention for purulent pancreatitis, have permitted to lower postoperative mortality from 26 to 10.5%. PMID- 10370309 TI - [The status of splanchnic blood circulation in patients with varicose veins of the esophagus and stomach in liver cirrhosis]. AB - Complex hemodynamical investigations were done in 166 patients with liver cirrhosis and the portal hypertension syndrome. Patients with varicose veins of the esophagus and stomach versus patients with isolated varicose veins of the esophagus have significantly higher resistance of vessels of the a. hepatica and v. porta systems, more pronounced losses of portal perfusion at the expense of varicose veins of stomach, gastro- and splenorenal shunts and lower volumetric blood flow in v. lienalis. While varicose veins of the esophagus and stomach occur an absolute values of the arterial and portal blood flow in liver are lowering, common hepatic blood flow reduces. The varicose veins of the stomach existence testifies high degree of the portosystemic shunting development with subsequent lowering of volumetric blood flow in v. lienalis in comparison with such in isolated varicose veins of the esophagus. PMID- 10370310 TI - [The forming of distal splenorenal anastomosis in conjunction with gastroesophageal junction suturing during treatment of the portal hypertension syndrome in children]. AB - The results of operations in 147 patients, performed for the portal hypertension syndrome, are adduced. The patients aged from 8 mo to 14 years. The best late follow-up result was noted after the distal splenorenal anastomosis forming in conjunction with suturing of esophageal veins and left gastric vein. PMID- 10370311 TI - [The iliac vein occlusion]. AB - Basing on roentgenological and morphological investigations the causes of the iliac veins passability disorder were studied up. In 45.1% of observations the stenosis was revealed of the left and in 1.2% of the right common iliac vein. Among the stenosis causes were osteal or cartilagenous prominences of vertebral column, intravascular structures (webs, membranes) in the left common iliac vein, the external iliac vein squeeze by internal iliac artery, retroperitoneal fibrosis, the right common iliac artery aneurysm, anomalous branching of parietal pelvic artery. PMID- 10370312 TI - [The use of extracorporeal stimulation of venous flow in the treatment of trophic ulcer in elderly and senile patients]. AB - Results of observations of 132 patients ageing from 61 to 93 years are adduced. There were performed the quilitative and quantitative examination of the trophic ulcer contamination with infection, comparative characteristics of coagulograms of a general blood flow and of the blood, out-flowing from trophic ulcer, prognostication and possibility of acceleration of the trophic ulcer healing depending on the organism reactivity using extracorporeal stimulation of venous outflow, its normalisation using the device for regulating compression of the body with the help of band dressings. PMID- 10370313 TI - [Immediate and remote results of cryosurgical treatment of oral cavity cancer]. AB - The results of treatment using cryodestruction, combined and complex therapy of the malignant tumor of the oral cavity mucosa in 116 patients was summarised. PMID- 10370314 TI - [Surgical treatment of postop large size abdominal hernia in patients with obesity]. AB - There were 140 patients with morbid obesity operated on for postoperative abdominal hernia. In 2 (1.4%) patients an acute cardiopulmonary insufficiency occurred, and in another 2 (1.4%) an acute thrombophlebitis of the lower extremities veins. Two patients died. The hernia recurrence have occurred in 3 (2.1%) patients operated according to Mayo method in terms from 1 to 5 years later. PMID- 10370315 TI - [Quantitative evaluation of severity degree of endotoxemia in burns and possibilities for its correction]. AB - Efferent and quantum methods were used in 176 injured persons with the burns and severe and extremely severe endotoxemia (ET) for additional nonspecific detoxication (AND). In 108 patients AND was applied for prophylaxis, beginning from the burn shock period, and in remaining patients--for significant ET, caused by the infection generalisation. An early sepsis was revealed in 3.7% of injured persons of the main and in 18.7% of control group. If the AND methods were applied while sepsis has begun in injured persons with extremely severe ET, the mortality was three times lower in comparison with such without AND application, causing the total mortality lowering in 2.7 times. PMID- 10370316 TI - [Surgical treatment of hemolytic anemia]. AB - Splenectomy was done in 448 patients with various forms of hemolytic anemia. Recovery was noted in 362 patients (of them 143 children) with hereditary microspherocytic hemolytic anemia (HMHA). Of 8 patients with hereditary nonmicrospherocytic hemolytic anemia (HNMHA) one died, in 7 general status had improved. Of 78 patients with autoimmune hemolytic anemia (AIHA) 3 died, in 47 (60%) satisfactory result was noted, general status improved--in 18 (23%). If surgical intervention is indicated the operation for HMHA is radical method of treatment, HNMHA--the selective one, and AIHA--one of the main. PMID- 10370317 TI - [Errors and hazards in the surgical treatment of vascular tumors of the mediastinum contributing to the respiratory tract compression in children]. AB - Examination and treatment of 16 children with vascular tumor of mediastinum ageing from 22 days to 11 years was conducted. In 12 patients the respiratory ways compression was noted. The mistakes and dangers while tumor excision were analysed, the methods of their prophylaxis using special technical tricks while performing the operation were elaborated. PMID- 10370318 TI - [The use of high-frequency artificial ventilation of lungs in the surgery of the trachea]. AB - In 100 patients the highly-frequential (HF) artificial pulmonary ventilation (APV) was applied while conduction of reconstructive operation on trachea and its bifurcation. While application of two-pulmonary ventilation or single-pulmonary and part of another pulmonary ventilation using HF APV an adequate pulmonary gas exchange was guaranteed, but using of HF APV of one pulmonary more than in 30% of observations severe hypercapnia and/or hypoxemia had occurred. In comparison with conventional methods of APV the improved conditions of surgeon's work appeared, the frequency of intraoperative complications--severe hypoxemia and hypercapnia- had reduced. The main fault of the HF APV method is the embarrassed monitoring of respiration efficacy. PMID- 10370319 TI - [The study of anastomoses performed with the utilization of synthetic absorbable sutures in the surgery of the stomach and intestines]. AB - Intestinal and gastro-intestinal anastomoses, performed using synthetic absorbable suture materials, foreign-dexon, vicryl and polydioxanon and native oktselon, were studied up in experiment. The oktselon thread do not cede to foreign threads in quality and has advantages over them in some features. It is possible to regulate the terms of a firmness loss and resorption changing the threads oxidation degree during their manufacturing. In tissues, sutured using oktselon thread, the processes of reparative regeneration are passing in maximally favourite conditions. PMID- 10370320 TI - [Current status of neurosurgical services in the Ukraine and prospects of their development]. PMID- 10370321 TI - [The main trends in the reform of the surgical services in the regional administration, improvement of the quality and efficacy of surgical services to the population]. PMID- 10370322 TI - [Modern approach to surgical treatment of duodenal ulcers]. PMID- 10370323 TI - [Sutureless methods of surgeries on gastrointestinal organs in clinic and experiment]. PMID- 10370324 TI - [Application of external endoscopic duodenotomy to duodenal sutures insufficiency]. PMID- 10370325 TI - [Follow-up study results of organ-salvaging surgeries for perforative pyloric duodenal ulcers]. PMID- 10370326 TI - [The combination of perforative gastric ulcer and hepatic and splenic laceration after closed abdominal trauma]. PMID- 10370328 TI - [Generalized type of fat embolism]. PMID- 10370327 TI - [X-ray-guided surgical revascularization of profound femoral artery in patients with arterial occlusive diseases of the lower extremities in a severe stage of ischemia]. PMID- 10370329 TI - Effects of response type on coordinated responses during arm movement. AB - This paper reports some experimental results on the coordination of finger and vocal responses with passing through a target position in multijoint arm movement. In Experiment 1, we found that the difference in the timing of finger and vocal responses cannot be attributed entirely to efferent or representational effects. Instead, it appears to reflect the extent to which information about the internal stimuli generated by the arm movement are available to the centers controlling these different responses. That is, it is a compatibility effect. In Experiment 2, the case in which a finger response is made on the same side of the body as the moving arm was compared with the case in which it is made with the contralateral hand, which remains static. The interaction effect observed suggests that the pathways subserving coordinated responses are informationally encapsulated, so that information about arm movement is not shared between the neural centers controlling different coordinated responses. PMID- 10370330 TI - Tactile roughness perception with a rigid link interposed between skin and surface. AB - Subjects made roughness judgments of textured surfaces made of raised elements, while holding stick-like probes or through a rigid sheath mounted on the fingertip. These rigid links, which impose vibratory coding of roughness, were compared with the finger (bare or covered with a compliant glove), using magnitude-estimation and roughness differentiation tasks. All end effectors led to an increasing function relating subjective roughness magnitude to surface interelement spacing, and all produced above-chance roughness discrimination. Although discrimination was best with the finger, rigid links produced greater perceived roughness for the smoothest stimuli. A peak in the magnitude-estimation functions for the small probe and a transition from calling more sparsely spaced surfaces rougher to calling them smoother were predictable from the size of the contact area. The results indicate the potential viability of vibratory coding of roughness through a rigid link and have implications for teleoperation and virtual-reality systems. PMID- 10370331 TI - Haptic estimates of discordant visual-haptic size vary developmentally. AB - In two size-conflict experiments, children viewed various squares through a reducing (1/2) lens while manually grasping them through a hand-concealing cloth. Then, using either vision or touch, they selected a match from a set of comparison squares. Forty 6-, 9-, and 12-year-olds participated in Experiment 1. Vision dominated the visual estimates of all three age groups; however, for the haptic estimates, the dominant modality varied developmentally: Vision dominated the 6-year-olds' haptic estimates, whereas neither modality dominated the 9-year olds' haptic estimates, and touch dominated the 12-year-olds' haptic estimates. In Experiment 2, 24 six-year-olds were tested, as in Experiment 1; however, half of them were shown the size-distorting effects of the lens just prior to testing. Although this reduced the visual dominance of their haptic estimates, the effect was weak and short-lived. The haptic dominance seen in the data of the 12-year olds was conspicuously absent. PMID- 10370332 TI - Contribution of ankle, knee, and hip joints to the perception threshold for support surface rotation. AB - The purpose of the present experiment was to investigate the extent to which subjects can perceive, at very slow velocities, an angular rotation of the support surface about the medio-lateral axis of the ankle, knee, hip, or neck joint when visual cues are not available. Subjects were passively displaced on a slowly rotating platform at .01, .03, and .05 deg/sec. The subjects' task was to detect movements of the platform in four different postural conditions allowing body oscillations about the ankle, knee, hip, or neck joint. In Experiment 1, subjects had to detect backward and forward rotation (pitching). In Experiment 2, they had to detect left and right rotations of the platform (rolling). In Experiment 3, subjects had to detect both backward/forward and left/right rotations of the platform, with the body fixed and the head either fixed or free to move. Overall, when the body was free to oscillate about the ankle, knee, or hip joints, a similar threshold for movement perception was observed. This threshold was lower for rolling than for pitching. Interestingly, in these postural conditions, an unconscious compensation in the direction opposite to the platform rotation was observed on most trials. The threshold for movement perception was much higher when the head was the only segment free to oscillate about the neck joint. These results suggest that, in static conditions, the otoliths are poor detectors of the direction of gravity forces. They also suggest that accurate perception of body orientation is improved when proprioceptive information can be dynamically integrated. PMID- 10370333 TI - Tops are more salient than bottoms. AB - Past research has verified that observers assume that objects are reliably oriented with respect to a gravitationally centered coordinate system. Observers also appear to attend more to specific parts of objects, like faces, that typically are closer to the top. In the present work, we explored whether or not observers have a generic bias to view tops as being more salient than bottoms. In three experiments, observers indicated whether random shapes appeared to be more similar to comparison shapes that shared identical tops rather than bottoms. Observers exhibited a reliable tendency to match figures with similarly shaped tops. Matching choice was also a function of global shape attributes such as axis of elongation or size. The findings are consistent with the notion that, in nature, tops tend to be the most visible part and to provide the best information with respect to important aspects of objects such as animal intentionality and artifact functionality. PMID- 10370334 TI - Parsing silhouettes: the short-cut rule. AB - Many researchers have proposed that, for the purpose of recognition, human vision parses shapes into component parts. Precisely how is not yet known. The minima rule for silhouettes (Hoffman & Richards, 1984) defines boundary points at which to parse but does not tell how to use these points to cut silhouettes and, therefore, does not tell what the parts are. In this paper, we propose the short cut rule, which states that, other things being equal, human vision prefers to use the shortest possible cuts to parse silhouettes. We motivate this rule, and the well-known Petter's rule for modal completion, by the principle of transversality. We present five psychophysical experiments that test the short cut rule, show that it successfully predicts part cuts that connect boundary points given by the minima rule, and show that it can also create new boundary points. PMID- 10370335 TI - Uniform connectedness and classical Gestalt principles of perceptual grouping. AB - We assessed whether uniform connectedness (UC; Palmer & Rock, 1994) operates prior to effects reflecting classical principles of grouping: proximity and similarity. In Experiments 1 and 2, reaction times to discriminate global letters (H vs. E), made up of small circles, were recorded. The small circles were respectively grouped by proximity, similarity of shapes, and by UC. The discrimination of stimuli grouped by similarity was slower than those grouped by proximity, and it was speeded up by the addition of UC. However, the discrimination of stimuli grouped by proximity was unaffected by connecting the local elements. In Experiment 3, similar results occurred in a task requiring discrimination of the orientation of grouped elements, except that the discrimination of stimuli grouped by UC was faster than that of those grouped by weak proximity. Experiment 4 further showed that subjects could respond to letters composed of discriminably separate local elements as fast as to those without separated local elements. The results suggest that grouping by similarity of shapes is perceived slower than grouping by UC, but grouping by proximity can be as fast and efficient as that by UC. PMID- 10370336 TI - Searching for impossible objects: processing form and attributes in early vision. AB - In five experiments, we investigated the extent to which form (shape) and metric attributes (three-dimensional, 3-D, orientation), both defined by relations between line elements, are processed in early vision. Search for a target defined by an abstract property of form (i.e., impossibility) was slow and serial. In contrast, search for a 3-D orientation target was considerably easier. Subsequent experiments suggest that this difference reflects the fact that 3-D orientation is derivable from localized sets of lines, whereas impossibility is an idiosyncratic property of the complete set of relations between lines. We conclude that only "gross" aspects of form are available in early vision as the complete set of line relations is not processed preattentively. However, localized processing of line relations is sufficient to derive 3-D orientation. PMID- 10370337 TI - The role of letter identity and letter position in orthographic priming. AB - Four experiments are reported investigating orthographic priming effects in French by varying the number and the position of letters shared by prime and target stimuli. Using both standard masked priming and the novel incremental priming technique (Jacobs, Grainger, & Ferrand, 1995), it is shown that net priming effects are affected not only by the number of letters shared by prime and target stimuli but also by the number of letters in the prime not present in the target. Several null results are thus explained as a tradeoff between the facilitation generated by common letters and the inhibition generated by different letters. Inhibition was significantly reduced when different letters were replaced by nonalphabetic symbols. Facilitation effects disappeared when the common letters did not have the same relative position in the prime and target strings, thus supporting a relative-position coding scheme for letters in words. PMID- 10370338 TI - Ultra-precise quantal timing: evidence from simultaneity thresholds in long-range apparent movement. AB - Conditions for the disappearance of long-range apparent movement were investigated. In an experiment on beta motion, critical interstimulus intervals (ISIs) of downward simultaneity thresholds for stimuli presented in continuous alternation were determined for exposure durations (EDs) varying from 3 to 160 msec. Each subject performed each test twice. Data were collected in three sessions, each for one of three angular separations (3 degrees, 6 degrees, and 12 degrees) and the full set of EDs. The distribution of critical ISIs collapsed across subjects, EDs, and angular separations shows sharp maxima at regular distances within a range of 0-110 msec ISI. Significant or near-significant peaks were found at ISIs of 5, 9, 22, 27, 43, 55, and 107 msec. Although mean critical ISIs shifted with spatial separation, no essential shift of the main maxima occurred. Evidence of a periodic modulation with a period duration of 4.5 msec was obtained from the distributions of differences between critical ISIs of the first tests and their replications, which exhibit extremely low standard deviations (< 10 msec). These results agree well with previous analyses (Geissler, 1987, 1992) that led to a taxonomic model of quantal timing, briefly summarized in this paper. Further consequences are discussed and related to earlier developments (Geissler, 1991, 1992, 1997). PMID- 10370339 TI - Can new objects override attentional control settings? AB - Previous research suggests that attentional capture by abrupt onsets is contingent on top-down attentional control settings. Four experiments addressed whether similar contingencies hold for capture elicited by the appearance of new perceptual objects. In a modified spatial cuing task, targets defined by abrupt onset or color were paired with distractors consisting of an abrupt brightening of an existing object or the abrupt appearance of a new object. In Experiments 1 and 2, when subjects searched for an onset target, both distractor types produced evidence of capture. When subjects searched for a color target, however, distractors produced no evidence of attentional capture, regardless of whether they consisted of a new perceptual object or not. Experiments 3-5 showed that the lack of distractor effects in the color-target condition cannot be accounted for by rapid recovery from capture. It was concluded that attentional capture by new objects is subject to top-down modulation by attentional control settings. PMID- 10370340 TI - Age similarities in the inertial properties of attention. AB - Adult age differences in the mode of allocation of visual attention were investigated, using a visual search task with a circular display containing one target letter and seven distractor letters. In two experiments, a total of 56 younger adults (M = 20 years) and 56 older adults (M = 66 years) searched for a target appearing with equal probability at one of two cued locations. The first cue appeared 115 msec before display onset, and the second cue appeared with display onset; distance between the two cued locations was varied. Target identification performance indicated that attention was inertial, in that reaction time for second-cued targets was related either to the area of the portion of the visual field containing possible target locations or to the mean path length of a serial self-terminating search. There were no age-related decrements in the allocation of visual attention. PMID- 10370341 TI - Examining the effect of practice on inhibition of return in static displays. AB - In a recent article, Weaver, Lupianez, and Watson (1998) reported that both object-based and location-based inhibition of return effects were reduced with practice. The present study was conducted to (1) replicate the reduction of inhibition of return with practice in single-session experiments with a variety of displays and responses and (2) to examine the notion that the reduction was, at least partly, due to habituation. However, no evidence for practice-related changes in the size of the inhibitory effect were found over a series of different inhibition of return experiments using static displays (using various numbers of target locations, types of keypress responses, and number of trials). Overall, the results suggest that inhibition of return is a robust phenomenon and may not, with static displays, be especially sensitive to practice effects. PMID- 10370343 TI - Been there, survived that. PMID- 10370342 TI - An area theorem for the same-different experiment. AB - If observers in a same-different experiment base their decisions on the absolute difference between observations on a trial, the area under the receiver operating characteristic equals the maximum proportion of correct decisions that an unbiased independent-observations observer could attain. Even though the differencing strategy is suboptimal, the area measure yields an index of optimal performance. PMID- 10370344 TI - Honesty is best. PMID- 10370345 TI - Government regulations ridiculous. PMID- 10370346 TI - MedBytes. PMID- 10370347 TI - Forum on ethics. Where is the outrage? An open letter to the speaker of the TMA House of Delegates. PMID- 10370348 TI - Medicare's new army. PMID- 10370349 TI - Wading in the gene pool. PMID- 10370350 TI - Talking with teens. PMID- 10370351 TI - Who decides? PMID- 10370352 TI - Preparing for 2000. PMID- 10370353 TI - Third-party funding of health care services for the uninsured of Tarrant County. AB - This study investigated how many citizens of Tarrant County do not have health insurance, what sources fund their health care, and how this level of funding compares with the privately insured population. Data for 1995 were obtained from various government agencies, local hospitals, local charitable organizations, and a survey of local private physicians. Medicare and Medicaid were considered to be forms of health insurance. Approximately 20% of the citizens of Tarrant County did not have health insurance. Third-party funding for health care for this population was approximately 56% of the funding for the privately insured population ($967 vs $1726 per person per year). The Tarrant County Hospital District contributed 52% of the uninsured funding; private hospitals and physicians, 26%; other state and local agencies, 21%; and charitable donations, 1%. I conclude that health care for the uninsured of Tarrant County is rationed compared with that of the privately insured population. PMID- 10370354 TI - The public health care of Central Americans in Houston. AB - In recent years, Americans have witnessed a marked change in the source of immigrant groups. Current immigrants are more likely to be former residents of the less developed world (nations such as Guatemala, Nigeria, India, and Viet Nam) than were earlier immigrants. In urban Texas, the influx of peoples from Central America is particularly striking and is largely a consequence of homeland political and economic instability. The new immigrants tend to be young and sexually active. We analyzed utilization patterns of Central Americans at our district health care facilities over 18 months and compared results with those of our non-Central American health care recipients. The 30,000 annual visits by Central Americans accounted for 4% of all visits. Disproportionately large amounts of care were given for sexually transmitted diseases and obstetric problems; conversely, small amounts were given for chronic illnesses, infectious diseases, acquired immunodeficiency syndrome, mental health problems, and adverse fetal outcomes. Few exotic tropical diseases were recognized or treated. PMID- 10370355 TI - Society for Epidemiologic Research 32nd annual meeting. Baltimore, Maryland, USA. June 10-12, 1999. Abstracts. PMID- 10370356 TI - Central nervous system cancers in first-degree relatives and spouses. AB - The increasing incidence of high-grade astrocytomas in the elderly, the associations between these malignancies and environmental factors, and case reports suggesting a familial component to these tumors prompted this study of primary brain tumors in first-degree relatives and spouses. This article describes the findings in 154 patients from 72 consecutive families accrued to the National Familial Brain Tumor Registry from 1991 to 1996. Medical records, pathological slides, and demographic data were reviewed for each identified case. Parents and children were affected in 33 families, siblings in 27, and husbands and wives in 12. The median age of the patients was 50.5 years, 55% were men, and 70% had high-grade astrocytomas. The pattern of tumor occurrence in this population is different from most familial cancers. These tumors did not involve multiple generations or occur at an unusually early age. In addition, the cases tended to cluster in time, with 47% of the familial and 50% of the husband-wife cases occurring within a 5-year span. In families with an affected parent and child, the diagnosis was made in the child before the parent in 45% of the cases. Prognostic factors for these patients appear to be similar to that reported for typical high-grade astrocytomas. This study demonstrates that primary brain tumors can occur in families without a known predisposing hereditary disease. The ages of these patients, the clustering of cases in time, the few affected generations, and the occurrence of brain tumors in spouses suggest that environmental exposures may be important in the etiology of this neoplasm. Although this hypothesis requires further study, it is plausible given the known associations in animals and humans between high-grade astrocytomas and radiation, toxic chemicals, and viruses. PMID- 10370357 TI - Randomized double-blind comparison of single high-dose ondansetron and multiple standard-dose ondansetron in chemotherapy-naive pediatric oncology patients. AB - This prospective, double-blind, randomized study compares the antiemetic efficacy of an equivalent dose of ondansetron administered as a single high dose or as multiple standard doses in pediatric oncology patients. Thirty-one chemotherapy naive patients were randomized at diagnosis to receive either single high-dose ondansetron (0.6 mg/kg, maximum dose 32 mg) or multiple standard-dose ondansetron (0.15 mg/kg, maximum dose 8 mg, every 4 hr for four doses). Antiemetic efficacy was assessed by an emesis scale described as follows: 1, no nausea or emesis; 2, nauseous but able to eat; 3, nauseous and unable to eat; and 4, emesis. Sixteen patients received high-dose and 15 received standard-dose ondansetron. Patients receiving moderately or severely emetogenic chemotherapy were evenly distributed between the two treatment groups. Eighty-one percent of patients receiving high dose and 80% receiving standard-dose ondansetron rated 1 or 2 on the emesis scale (p = 0.93). No patient experienced any clinical or laboratory toxicity. Our study suggests that single high-dose ondansetron is as efficacious as the multiple standard-dose regimen and is well tolerated. Its use will facilitate the administration of ondansetron in pediatric patients receiving chemotherapy. PMID- 10370358 TI - Activities of DNA turnover and free radical metabolizing enzymes in cancerous human prostate tissue. AB - Activities of adenosine deaminase (ADA), 5'nucleotidase (5'NT), xanthine oxidase (XO), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) and levels of thiobarbituric acid reagent substances (TBARS) were measured in 10 cancerous and 10 noncancerous human prostate tissues. Decreased activities of DNA turnover enzymes (ADA and 5'NT), increased activities of GSH-Px and CAT, and unchanged activities of SOD and XO were observed in cancerous prostate tissues compared with those of noncancerous ones. TBARS levels were found to be higher in cancerous tissues than noncancerous ones. In correlation analysis, mostly positive correlations were established between enzyme activities of the cancerous tissues, whereas no meaningful correlations were found between enzyme activities of the noncancerous tissues except for a positive correlation between XO and SOD. The results indicate that the activities of DNA turnover enzymes were reduced, which was possibly an attempt to lower the rate of purine catabolism, and the activities of GSH-Px and CAT enzymes were increased, probably in response to increased free radical stress occurring in cancerous prostate tissues. Increased concentrations of TBARS suggested oxidant stress and thus accelerated peroxidative reactions in the cancerous tissues, even though antioxidant defense mechanisms were activated. These findings suggest that enzymatic antioxidant systems of cancerous prostate tissues cannot sufficiently eliminate oxidant factors and prevent cellular peroxidative reactions occurring during the carcinogenic process. PMID- 10370359 TI - Treatment of a malignant enterocutaneous fistula with octreotide acetate. AB - An enterocutaneous malignant fistula developed in a patient who had a retroperitoneal angiosarcoma. He was treated with octreotide acetate subcutaneously. Drainage decreased and ceased after 2 weeks of therapy. The closure of this malignant fistula suggests that palliative therapy with octreotide acetate merits further study in view of the grave prognosis of this complication. PMID- 10370360 TI - A thousand points of light or just dim bulbs? Radiolabeled antibodies and colorectal cancer imaging. AB - Radioimmunoscintigraphy (RIS) is coming into its own as an imaging modality in clinical oncology. Early experience with indium-111-labeled intact murine monoclonal antibodies (MoAbs) in colorectal cancer suggested that RIS images hepatic metastases poorly. Moreover, an antimurine immune response was frequently provoked, precluding multiple follow-up RIS studies in individual patients due to reticuloendothelial sequestration of the radioimmunoconjugate before tumor targeting could occur. Recent trials of technetium-99m-labeled antibody fragments and human MoAbs have demonstrated significant improvement in imaging efficacy, and repeated or serial imaging is possible because of the absence of associated immunogenicity. RIS is demonstrably more sensitive than conventional diagnostic modalities (CDM) such as computed tomography (CT) for detection of extrahepatic abdominal and pelvic colorectal carcinoma and is complementary to CDM in imaging liver metastases. In a surgical decision-making analysis comparing CT, RIS (IMMU 4 99mTc-Fab'; CEA-Scan), and CT plus RIS in patients with recurrent or metastatic colorectal cancer, CT plus RIS improved correct prediction of resectability by 40% and correct prediction of unresectability by 100% compared with CT alone. At the present time, RIS used in combination with CDM contributes an incremental improvement in diagnostic accuracy in colorectal cancer patients with known or suspected recurrent disease. Basic and clinical research currently in progress promises to yield agents and methods that provide rapid high-resolution imaging, high tumor-to-background ratios in all organs at risk for tumor recurrence or metastasis, negligible immunogenicity and toxicity, and a significant further improvement in the accuracy of clinical decision making in oncology patients. PMID- 10370361 TI - Psychological aspects of ovarian cancer. AB - Ovarian cancer presents a range of physical and psychological symptoms during stages of diagnosis, treatment, and survival. Women at risk for ovarian cancer who attend screening programs are vulnerable to high levels of depression and anxiety, particularly young women with poor social support. Multiple physiological stressors of surgical menopause, steroid therapy, and pain present during active treatment that place women at high risk of depression and anxiety during this time. Symptoms of anxiety and depression are also prevalent immediately after chemotherapy and during palliative care. Screening for psychological distress may be useful to identify women who will benefit from psychological counseling. They should be referred to a mental health professional affiliated with the hospital at which they are receiving oncology services. Brief group or individual supportive psychotherapies are effective in relieving psychological distress. Face-to-face psychological intervention should be tailored to the patient's degree of physical mobility. Pain, discomfort, and severe mood symptoms should be addressed pharmacologically, when possible, by a psychiatric consultant knowledgeable in oncology psychiatry. Survivors experience chronic fear of recurrence, sexual dysfunction, and identity disturbance. Reports that ovarian cancer can result in positive life changes, such as closer interpersonal relationships, are encouraging and may provide hope to patients who become despairing about the future. PMID- 10370363 TI - Narrowing the options: the process of deciding on prostate cancer treatment. AB - Prostate cancer is a pressing health concern in the United States and one surrounded by continual controversy. Currently there is no consensus regarding the efficacy of routine screening, nor has one treatment modality been demonstrated as superior. Patients and spouses are asked to choose from several options: radical prostatectomy, radiation therapy, or the "watch and wait" approach. A grounded theory design was used to examine the actual treatment decision-making process as it occurred over time among 18 newly diagnosed prostate cancer patients and their wives in western North Carolina. Couples were interviewed conjointly and individually to explore their perceptions of the decision process. All interviews were audiotaped, transcribed verbatim, and analyzed using content analysis techniques. Couples negotiated decisions through their common and unique personal and family histories, biases, and individual coping styles. They narrowed the options based on these factors. Most couples received their counseling regarding treatment options exclusively from the surgeon who narrowed the options based on age and physiologic status. Most couples chose surgery believing it to be the only treatment promising cure. Distinct misconceptions about radiation therapy were noted. Concern about potential side effects did not deter men from selecting surgery, although men and their wives differed in their willingness to accept treatment "at any cost." Information regarding potential for cure and risk of recurrence were highly important factors in the decision process. The decision was incomparable with any other life decisions the couples had faced. PMID- 10370362 TI - Psychosocial concerns and quality of life in breast cancer survivors. PMID- 10370365 TI - Growth of oncology physician practice management companies. AB - The practice of oncology is changing dramatically, spurred on by managed care initiatives throughout the United States. As a result, physicians are faced with multiple demands from insurers, managed care organizations, and patients. In response to these demands, oncology physician practice management companies have entered the cancer market. This article describes the driving factors leading to consolidation in practice settings, the risks and benefits to oncologists of affiliating with these companies, and the organizational characteristics of four of these larger corporations. This review article is of broad interest to oncologists practicing in the United States and is meant to provide a useful reference for considering a physician practice management company as a business partner. PMID- 10370366 TI - New chemoimmunotherapy: courtesy of a more flexible Food and Drug Administration. PMID- 10370367 TI - Decision to operate, radiate, or watch and wait: the prostate cancer dilemma. PMID- 10370368 TI - Free radicals, oxidative stress, and DNA metabolism in human cancer. PMID- 10370369 TI - Bruton's tyrosine kinase controls a sustained calcium signal essential for B lineage development and function. AB - Genetic data support a role for Btk during the B lineage development transitions regulated by signaling through both the pre-B and the B cell antigen receptors. Dysregulated signaling at each of these transitions can result in failure of these cell populations to proliferate and subsequent cell death. Btk-dependent IP3 production is crucial for maintaining the sustained calcium signal in response to BCR engagement and is likely to regulate a subset of transcriptional events essential for B lineage growth or survival. Identification of these Btk dependent signals will be important in understanding B cell activation, differentiation, and cell death. This information may lead to therapies specifically targeting these events in B cell autoimmunity or malignancy and provide a fuller understanding of the appropriate target populations and potential negative consequences of Btk gene therapy in XLA. Identification of Btk/Tec family kinases in an increasing number of vertebrate and invertebrate cell lineages suggests that the link between Btk and the PLC gamma/IP3/calcium signaling pathways may be broadly conserved. PMID- 10370370 TI - Regulation of CCR5 and CXCR4 expression by type 1 and type 2 cytokines: CCR5 expression is downregulated by IL-10 in CD4-positive lymphocytes. AB - HIV-1 transmission and disease progression is, in general, characterized by initial predominance of macrophage tropic, non-syncytium-inducing strains followed by a switch to T-cell tropic, syncytium-inducing strains. Using sensitive, quantitative kinetic RT-PCR, we examined cytokine regulation of tropism-specific HIV-1 coreceptor expression in PBMCs from HIV-1-seronegative individuals. Proinflammatory (TNF-alpha and IL-12) and type 1 cytokines (IFN gamma and IL-2) significantly upregulated CCR5 (wt allele) mRNA expression in CCR5 homozygous wild-type (wt/wt) and heterozygous individuals (wt/del) (P < 0.02). CCR5 (wt) mRNA expression in unstimulated PBMCs was significantly increased in wt/wt individuals compared to that of wt/del individuals (P < 0.01). In wt/del individuals, del CCR5 mRNA was expressed at 10-fold greater levels than wt CCR5 mRNA in unstimulated PBMCs from the same individual. Flow cytometry confirmed that upregulated CCR5 mRNA following type 1 cytokine stimulation leads to increased cell surface expression of CCR5 protein. The type 2 cytokine IL-10 downregulated both CCR5 mRNA and protein expression in wt/wt and wt/del individuals. Proinflammatory, type 1, and type 2 cytokines significantly increased CXCR4 mRNA expression in wt/wt, wt/del, and del/del CCR5 genotypes (P < 0.02). These results suggest that changes in the cytokine milieu influence chemokine receptor expression and may explain emergence of tropism-specific strains facilitating HIV transmission and disease progression. PMID- 10370371 TI - Discordance between IgA switching at the DNA level and IgA expression at the mRNA level in IgA-deficient patients. AB - IgA deficiency is a common immune disorder in Caucasians and is associated with certain MHC conserved extended haplotypes, such as [HLA-B8, SC01, DR3], which presumably carry a susceptibility gene(s). We applied a competitive digestion circularization PCR method to quantitate the number of switch (S)mu to S alpha rearrangements in peripheral B cells from IgA-deficient subjects homozygous for this haplotype and compared their number with the productive C alpha mRNA level to determine C alpha gene expression in IgA-switched B cells. Two types of defects, low expression of both secreted and membrane forms of productive C alpha mRNA in IgA-switched B cells and impaired IgA switching, were characterized in IgA-deficient subjects homozygous for [HLA-B8, SC01, DR3]. The former defect was also found in another noncarrier subject. It may directly cause low IgA secretion and reflects a blockade in post-IgA switch differentiation of B cells. These results suggest that the heterogeneity of defects in IgA deficiency is not simply ascribable to MHC susceptibility genes. PMID- 10370373 TI - Decreased dipeptidyl peptidase IV enzyme activity of plasma soluble CD26 and its inverse correlation with HIV-1 RNA in HIV-1 infected individuals. AB - Human plasma contains soluble CD26/dipeptidyl peptidase IV (sCD26/DPPIV) although its physiological significance remains unclear. To determine whether the plasma sCD26 levels have clinical relevance in HIV-1 infected individuals, the concentration and DPPIV enzyme activity of plasma sCD26 were measured. While there is no significant difference between the plasma levels of sCD26 in 90 HIV-1 infected individuals and in 79 uninfected controls, specific DPPIV enzyme activity of sCD26 was significantly decreased HIV-1 infected individuals (P < 0.0001). Specific DPPIV enzyme activity was correlated with the levels of CD4+ T cells (r = 0.247; P < 0.02), CD8+ T cells (r = 0.236; P < 0.03), and adenosine deaminase (r = 0.227; P < 0.05) and had an inverse correlation with HIV-1 RNA (Spearman's r = 0.474; P = 0.0012). Furthermore, recombinant sCD26 enhanced the in vitro PPD-induced response of lymphocytes from HIV-1 infected individuals with decreased specific DPPIV enzyme activity. These results suggest that the specific DPPIV enzyme activity of plasma sCD26 may contribute to the immunopathogenesis of HIV infection. PMID- 10370372 TI - C/EBP beta in rheumatoid arthritis: correlation with inflammation, not disease specificity. AB - Rheumatoid arthritis synovial tissue was examined and compared with osteoarthritis tissue for the presence of the nuclear transcription factor C/EBP beta (NF-IL-6). The region (lining or sublining), cell type, and subcellular distribution (cytoplasmic or nuclear) of the expression of C/EBP beta was characterized. Rheumatoid arthritis synovial fluid and blood and normal peripheral blood were also examined. C/EBP beta was detected in the synovial lining and in sublining cells of synovial tissue from patients with both rheumatoid and osteoarthritis. A significant (P < 0.001 and < 0.05, respectively) increase in the percentage of cells with nuclear staining was seen in the lining layer, compared to cells in the sublining region, in rheumatoid and osteoarthritis. In both diseases a strong correlation (r = 0.79, P < 0.001) was observed between the percentage of cells in the synovial lining that were positive for nuclear C/EBP beta and lining cell depth. Two-color immunohistochemistry demonstrated that both macrophages and fibroblast-like synoviocytes were positive for nuclear C/EBP beta. The presence of C/EBP beta was confirmed by immunohistochemistry and Western blot analysis with isolated synovial fibroblasts. Nuclear C/EBP beta was also detected in rheumatoid synovial fluid monocytes/macrophages, but not in lymphocytes or neutrophils. Western blot analysis confirmed the presence of C/EBP beta in these cells. The intensity of C/EBP beta staining was greater (P < 0.001) in synovial fluid monocytes than in those from normal or rheumatoid peripheral blood. In conclusion, the enhanced nuclear staining for C/EBP beta in the synovial lining, compared to the sublining, suggesting activation in the lining, and the positive correlation of lining layer depth with the percentage of cells in the lining positive for nuclear C/EBP beta, suggest a potential role for C/EBP beta in chronic inflammation. The regulation of the production or activity of C/EBP beta, to inhibit inflammatory mediator expression by synovial macrophages and fibroblasts, offers a novel approach to therapeutic intervention. PMID- 10370374 TI - Correlation of thymic pathology with HLA in myasthenia gravis. AB - The aim of this study was to investigate associations between thymic pathology and HLA in myasthenia gravis. HLA typing was performed in 95 of 125 Caucasian patients who underwent transsternal thymectomy for myasthenia gravis between 1976 and 1995. Multiple comparison procedures applied within each HLA locus demonstrated significant correlations between the ancestral suprahaplotype A1 B8 DRB1*0301 DRB3*0101 DQA1*0501 and thymic hyperplasia and between HLA-A24 and thymoma. A weaker association was found between A3 and thymic atrophy and thymolipoma. On logistic discriminant analysis, HLA-B8 (P = 0.001) and HLA-A3 (P = 0.028) were identified as the only significant classifiers to jointly provide a good discriminator between the thymic pathologies. When the suitability of HLA for detection of thymoma was examined in a second logistic regression analysis, both HLA-A24 (OR 9.7; 95% CI [1.6, 73.7]) and HLA-B8 (OR 0.1; 95% CI [0.0, 0.5]) were significant predictive factors. The above correlations between thymic pathology and HLA-A3, HLA-A24, and HLA-B8 (but not MHC class II alleles) suggest an involvement of MHC class I restricted T cells in myasthenic autoimmunity that may partially be reflected by thymic pathology. PMID- 10370375 TI - Ectopic expression of B7-1 (CD80) on T lymphocytes in autoimmune lpr and gld mice. AB - Defective Fas-mediated apoptosis in mice, caused by the gld mutation in the fas ligand gene, results in the development of lupus-like autoantibodies and severe lymphoproliferation. We previously demonstrated ectopic expression of the costimulatory molecule B7-1 (CD80) on T lymphocytes in B6/gld mice. This report extends these observations by demonstrating similar results in B6/lpr mice, which possess a mutation in the gene encoding Fas. Additionally, we demonstrate that this phenomenon is age-dependent and occurs on multiple subsets of B6/gld T lymphocytes. B7-1 upregulation is observed on T cells from both conventionally housed and specific-pathogen-free B6/gld mice, suggesting that this is not a consequence of infection by pathogen. T cells from lpr and gld mice show increased binding of CTLA4-Ig fusion protein, suggesting that the upregulated B7 1 is functional. CD28, a receptor for B7-1 which activates T cells, is upregulated in B6/lpr and B6/gld mice, while CTLA4, a negative regulator of T cells which binds B7-1, is not. Our results suggest that ectopic expression of B7 1 on T cells of lpr and gld mice may be playing a role in exacerbation of lymphoproliferation and/or autoimmunity. PMID- 10370376 TI - Protection from Lyme neuroborreliosis in nonhuman primates with a multiantigenic vaccine. AB - In an effort to develop an effective and safe vaccine for lyme disease, rhesus macaques were injected with a multiantigenic preparation of Borrelia burgdorferi, strain N40. One month later animals were boosted before intradermal challenge with infectious B. burgdorferi. Challenges were performed at 1 and again at 5 months after the booster vaccination. Vaccinated and control nonvaccinated animals were monitored for development of systemic infection by measurement of serum anti-spirochetal antibodies by ELISA and Western blotting, and neurological involvement was monitored by testing of cerebrospinal fluid (CSF) and PCR analysis of central nervous system (CNS) tissue obtained at necropsy. Two control nonhuman primates (NHPs), given saline injections instead of vaccine and then challenged with B. burgdorferi, developed CSF pleocytosis, PCR positivity of the brain, and high levels of specific anti-B. burgdorferi antibody in the serum and CSF. Necropsy studies revealed widespread invasion of the CNS of one of these animals by the spirochete. In contrast, none of the four vaccinated animals developed evidence of invasion of the CNS after either of two challenge inoculations with infectious B. burgdorferi. In addition to resisting infection, no vaccinated animal demonstrated any untoward consequence of vaccination. These data demonstrate that a multiantigenic vaccine is effective in preventing systemic infection and lyme neuroboreliosis in NHPs and suggest that a successful vaccine could be developed in humans which would prevent lyme disease. PMID- 10370377 TI - Characterization of the local and systemic immune responses in patients with cutaneous leishmaniasis due to Leishmania major. AB - In this study skin biopsies and peripheral blood samples were obtained from patients with cutaneous leishmaniasis caused by Leishmania major. Samples were obtained at diagnosis and during healing when the lesions had regressed to half the original size. At diagnosis most of the cells expressed HLA-DR. The numbers of CD8+ cells in the lesions were higher at diagnosis than during healing. By contrast, a lower percentage of PBMC expressed CD8+ cells at diagnosis probably due to sequestration in the lesion. In the lesion, in situ staining for IFN gamma, IL-10, and IL-4 showed marked variation between all patients in the number of positive cells for a particular cytokine. The proliferative response of PBMC to leishmanial antigens and IFN-gamma production tended to increase during healing. Cytokine patterns in the PBMC in response to Leishmania antigen was more specific than in the lesion and correlated better with the clinical manifestations. Possible reasons for this are discussed. PMID- 10370378 TI - Effects of SIVmac infection on peripheral blood CD4+CD8+ T lymphocytes in cynomolgus macaques. AB - We have previously reported that CD4+CD8+ double-positive (DP) T cells with a resting memory phenotype exist in a substantial proportion of peripheral blood lymphocytes of adult cynomolgus macaques. In this study, we examined the effects of simian immunodeficiency virus of macaque (SIVmac) infection on DP T cells. In vitro, SIVmac239 nef-open (239) and its nef-deletion mutant replicated well in both CD4+CD8- and DP T cells. However, when the macaques were infected with 239, DP, but not CD4+CD8-, T cells were transiently increased in parallel with cell activation and viral replication, followed by depletion within 1 month postinfection. Interestingly, the nef gene was required for depletion but not for the increase and activation of DP T cells. These data suggest that the pathogenic SIV infection may downmodulate production and/or blood circulation of DP T cells by a Nef function-related mechanism(s) different from that for the depletion of CD4+CD8- T cells. PMID- 10370379 TI - Enhancement of endothelial cell E-selectin expression by sera from patients with active Behcet's disease: moderate correlation with anti-endothelial cell antibodies and serum myeloperoxidase levels. AB - We studied the in vitro E-selection expression of endothelial cells treated with sera from patients with Behcet's disease (BD) and factors (anti-endothelial cell antibodies, anti-neutrophil cytoplasmic antibodies, cytokines, and myeloperoxidase (MPO) that may contribute to adhesion molecule expression. A total of 21 patients with BD and 27 healthy controls were studied. In vitro E selectin endothelial cell expression was investigated by ELISA after HUVEC incubation with sera or purified IgG from patients with BD and controls. Increased E-selectin expression was observed when endothelial cells were incubated with sera from patients with active disease or from patients with circulating anti-endothelial cell antibodies and high levels of plasma myeloperoxidase. Abnormalities of endothelial cell function have been suggested to play a role in the occurrence of vascular damage in BD. Our findings suggest that anti-endothelial cell antibodies and neutrophil hyperactivity, as inferred from the high plasma MPO levels in patients with active disease, may explain endothelial cell activation and neutrophil accumulation in vascular lesions. PMID- 10370380 TI - Phenotype and cytokine profile of Schistosoma mansoni specific T cell lines and clones derived from schistosomiasis patients with distinct clinical forms. AB - It is essential to distinguish the role of T lymphocytes on the physiopathology associated to more severe forms of schistosomiasis and on the immunomodulation that evolves in the majority of infected people. In this study, we generated Schistosoma mansoni-specific T cell lines and clones from patients with the acute and chronic (intestinal and hepatosplenic forms) phases of disease, from former ones, and from uninfected individuals sensitized to parasite soluble antigens. T cell lines derived from nontreated acute infected donors were capable of producing IL-4 and IL-5, while cells from treated patients secreted IFN-gamma. Lines from intestinal chronic and antigen-sensitized donors preferentially produced IFN-gamma, while those from hepatosplenic patients secreted all three cytokines. The cytokine analysis of CD4+ T cell clones revealed a Th2/Th0 pattern (clones producing IL-4 and IL-5 and clones producing all three cytokines) for those derived from infected patients, while cells from antigen-sensitized donors exhibited an opposite Th1/Th0 pattern (clones producing IFN-gamma and clones producing all three cytokines). The possible role of these T cell populations on human schistosomiasis mansoni is discussed. PMID- 10370381 TI - Expression of costimulatory molecules CD80 and/or CD86 by a Kaposi's sarcoma tumor cell line induces differential T-cell activation and proliferation. AB - During physiological stimulation of resting T-cells, at least two activation signals by antigen presenting cells are required. Besides the first antigen specific signal, the second costimulatory signal involves CD80 and CD86 expressed by the antigen presenting cell. These costimulatory molecules have been suggested to be of clinical relevance in many different autoimmune and malignant disease processes. We previously observed that tumor cells in Kaposi's sarcoma (a common AIDS-related cutaneous neoplasm) completely lack both CD80 and CD86, and these tumor cells fail to stimulate T-cell proliferation. In this study, using a Kaposi's sarcoma tumor cell line designated SLK, various stable transfected cell lines were produced. Tumor cells that were either singly positive for either CD80 or CD86, as well as a double-positive cell line, were examined for their ability to induce T-cell activation, T-cell proliferation, and cytokine production profiles. Compared to the parental double-negative tumor cell line, the CD80 positive cells, but not the CD86-positive tumor cells, induced significant T-cell activation and proliferation. Tumor cells expressing both CD80 and CD86 also induced T-cell activation. After stimulation by the transfected tumor cells, T cells produced a Th-1 type cytokine production profile with increased IL-2 and IFN-gamma levels. These results demonstrate that Kaposi's sarcoma tumor cells lacking co-stimulatory molecules cannot induce T-cell activation; however, if they express CD80, they can induce peripheral blood T-cell proliferation, and there is a differential response as expression of CD86 did not have the same immunostimulatory effect. PMID- 10370382 TI - Neutrophils from Mycobacterium avium-infected mice produce TNF-alpha, IL-12, and IL-1 beta and have a putative role in early host response. AB - Recent evidence supports a role for neutrophils in the host defense against Mycobacterium avium. To determine whether the depletion of neutrophils has an effect on the outcome of infection in mice as determined by the number of bacteria in liver and spleen, we administered RB6-8C5 anti-neutrophil antibody intraperitoneally both early and late in the infection. Mice were then observed for 14 days and harvested. The number of viable bacteria in liver and spleen was determined. While administration of RB6-8C5 antibody early in infection resulted in a significant increase in the number of bacteria in organs when compared with mice receiving immunoglobulin control, administration of RB6-8C5 antibody late in infection (week 3) did not have an impact on the bacterial load in tissue. Infection of CD18 knockout mice (with impaired neutrophil function), however, did not show a significant enhancement of M. avium growth when compared with that of wild-type control mice. Neutrophils were found to produce increased amounts of TNF-alpha and IL-12 and IL-1 than control uninfected mice during the initial phase of infection, but not after 2 weeks following infection (although IL-1 beta levels continue elevated). The results suggest that neutrophils may have a role in the early (innate) immune response against M. avium but it is only evident after acute depletion of neutrophils and not in mice with chronic neutrophil impairment. PMID- 10370383 TI - [Electrophysiological brain-stem diagnosis in patients with a pronounced obstructive sleep apnea syndrome]. AB - BACKGROUND AND OBJECTIVE: The pathophysiological basis of obstructive sleep apnoea syndrome (OSAS) is a loss of tone of the oropharyngeal muscles during sleep, leading to partial and/or total collapse of the oropharyngeal muscle tube. The normal inspiration-synchronous activation (ISA) of the tongue muscles is diminished or lost in patients with OSAS. Observations of OSAS in patients with ischaemic pontomedullary lesions, syringobulbia (syringomyelia), olivo-ponto cerebellar atrophy or disseminated encephalopathy have indicated that it is due to impairment of ISA. In this study systematic electrophysiological studies (EPS) were undertaken in patients with severe OSAS, as demonstrated by polysomnography, to determine whether, in addition to the central abnormality of oropharyngeal muscular tone, one can demonstrate an increased incidence of subclinical, functional disorder in the brain stem as well as any pointers to a central localization of abnormalities in OSAS. PATIENTS AND METHODS: 19 patients with severe OSAS were examined for the presence of subclinical brain-stem lesions by means of acoustic evoked potentials, blinking reflex, masseter reflex, masseter silent period, electronystagmogram together with vestibular testing and magnetic evoked potentials of the tongue musculature. RESULTS: Findings were unremarkable in 15 of the 19 patients. EPS gave reproducible evidence of functional left pontine (blink reflex) brain-stem lesions in two of the patients and of bilateral pontomesencephalic (masseter reflex) brain-stem lesions in the other two. CONCLUSION: In the absence of systematic abnormalities our findings speak against relevant brain-stem lesions being the cause and/or consequence of OSAS. PMID- 10370384 TI - [Severe pulmonary hypertension in systemic lupus erythematosus. The successful therapy of an unusual manifestation]. AB - OBJECTIVE AND CLINICAL FINDINGS: A 48-year-old woman was hospitalized because of haemoptysis. Until shortly before admission she had been on phenprocoumon after pulmonary embolism sustained 18 months previously. Six months before admission systemic lupus erythematodes (SLE) had been diagnosed and treatment with cortisone initiated. Physical examination revealed jugular venous congestion, tachycardia, dyspnoea on even minimal physical activity and pretibial oedema. INVESTIGATIONS: Lung scintigraphy showed a perfusion deficiency in the right lung, unchanged since a test 18 month before. Doppler echocardiography recorded an estimated pulmonary artery systolic pressure of 110 mm Hg. Angiography showed a fully patent superior vena cava and nearly complete occlusion of the main right pulmonary artery by a thrombus. DIAGNOSIS, TREATMENT AND COURSE: The haemoptyses ceased after 5 days of treatment with methylprednisolone, 130 mg daily for 5 days, reduced after 5 days to 80 mg i.v. every other day, plus cyclophosphamide, 50 mg daily by mouth. The pulmonary hypertension remained unchanged so that pulmonary thrombendarterectomy was indicated. Surgery revealed extensive mediastinal fibrosis and almost complete occlusion of the thick-walled right pulmonary artery by thrombus adherent to the wall. Histology showed vasculitis of the pulmonary arterial intima and of the small pulmonary vessels. After thrombectomy the pulmonary arterial systolic pressure fell to an remained at below 40 mm Hg. Phenprocoumon was continued (at an INR of 2.5-3.5) as was immunosuppressive treatment. The patient has remained free of symptoms and is able to be physically active. CONCLUSION: Pulmonary hypertension is a serious complication of SLE. Echocardiography is recommended for both the original diagnosis and serial follow-up, complemented by other imaging methods if indicated. PMID- 10370385 TI - [Littoral-cell angioma--a rare differential diagnosis on splenic tumors]. AB - HISTORY AND PRE-ADMISSION FINDINGS: Routine abdominal sonography of a 51-year-old man 6 years after removal of the right testis and radiotherapy for a seminoma revealed a 3 cm mass within the spleen. INVESTIGATIONS: All biochemical tests were normal. Computed tomography (CT) and magnetic resonance imaging confirmed the tumour which had not been present on the CT before the seminoma had been treated. No other space-occupying lesions were found in the thorax and abdomen. TREATMENT AND COURSE: A splenectomy was performed because a metachronous metastasis of the seminoma was suspected. The operation and subsequent course were uneventful. At operation the tumour had been diagnosed as an haemangioma because of its gross appearance, but histological and immunohistochemical examination revealed a littoral cell angioma. CONCLUSION: The littoral cell angioma is a benign vascular lesion in the red pulp of the spleen, which may be caused by different stimuli such as chronic infection or tumours. This case illustrates, that this tumour should be considered in the differential diagnosis of an unclear neoplasm in the spleen. PMID- 10370386 TI - [Sleep apnea and erectile dysfunction]. PMID- 10370387 TI - [Interferon-alpha and ribavirin: advances in the therapy of chronic hepatitis C]. PMID- 10370388 TI - [The object and goals of rehabilitation. Rehabilitation in health insurance under law]. PMID- 10370390 TI - [Clinical research (3). Therapeutic studies]. PMID- 10370389 TI - [Depressive diseases after heart operations]. PMID- 10370391 TI - Scandinavian Society for the Study of Diabetes 34th annual meeting jointly with the Diabetes Education Study Group, Finland 1st annual symposium. Turku, Finland, 6-9 May 1999. Abstracts. PMID- 10370392 TI - Myocardial electrogenesis and the electrocardiogram. AB - Determinants of the electrocardiographic voltage are reviewed with formulation of certain parameters. The dipole moment of a single fiber and consequently, a possible maximal moment of the double layer of the activation wave front are estimated as 0.21 mA.cm per unit area (cm2). The longitudinal activation of parallel fibers produces much stronger double layer than the transverse activation across fibers. Without any loss of the electrical force of fibers, a normal ventricle of 200 g in weight would create a possible maximal QRS area of 350 microV.sec in a surface lead. The normal endo- to epicardial ventricular activation is predominantly transverse with respect to fiber orientation and rather inefficient in electrogenesis. It is implied that abnormalities in ventricular conduction may possibly improve the effectiveness of myocardial generator. PMID- 10370394 TI - Nocturnal oxygen desaturation in coronary artery disease. AB - Nocturnal oxygen desaturation and sleep apnea may provoke myocardial ischemia and arrhythmias in patients with coronary artery disease (CAD). Additionally, these factors may accelerate coronary atherosclerosis in the long term and they may play a role in the progression of the disease process. On the other hand, studies related to this subject are limited. This study was conducted to investigate the nocturnal oxygen desaturation and apneas during sleep in patients with CAD and to assess the possible association of these factors with CAD. We studied 22 male patients with CAD confirmed by coronary angiography who did not have symptomatic pulmonary disease and fourteen male healthy controls without known heart disease. Patients were randomly selected from men undergoing coronary angiography. Controls were age and sex matched and selected from the population registry. The normal controls were of similar body mass index to the patients. None of them were obese. The patients and controls underwent standard polysomnography. Men with CAD and controls had a similar apnea-hypopnea index (2.3 +/- 3.8 vs. 1.2 +/- 1.7). Mean oxygen desaturation index was higher among patients than controls (2.1 vs. 0.5, p < 0.05). Patients with CAD spent 3.1% (9.7 +/- 13.6) of total sleep time desaturated, while the same proportion in controls were 0.5% (1.9 +/- 4.1)(p < 0.05). Although both groups of patients were of similar heart rates at initial, the development of bradycardia during sleep was significantly higher in patients compared with controls (43.3% vs. 25.3%, p < 0.05). The results demonstrate that sleep disordered breathing, in particular nocturnal oxygen desaturation, occurs more common in patients with CAD compared to controls. Additionally, patients are at higher risk of developing bradycardia during sleep. This findings suggest that oxygen desaturation during sleep might contribute to the progression of CAD. PMID- 10370393 TI - Influence of calcium antagonists on long-term survival of patients treated with coronary angioplasty. AB - A meta-analysis reported that nifedipine increased mortality dose-dependently in patients with coronary artery disease. However, there have been few studies (specifically in Asians) on the long-term prognosis of patients treated with calcium antagonists after successful coronary angioplasty (PTCA). The subjects consisted of 583 consecutive patients (461 males, aged 59 +/- 10), who underwent successful elective PTCA between 1985 and 1990. First, they were divided into two groups; the calcium antagonist (+) group (n = 560) and the calcium antagonist (-) group (n = 23), and were evaluated in terms of total survival and cardiac events. Second, the calcium antagonist (+) group was further divided into 4 groups according to calcium antagonist type, i.e., short-acting nifedipine group (n = 156), long-acting nifedipine group (n = 203), diltiazem group (n = 184) and the other group (n = 17), and these groups were evaluated in the same way. The primary end-point was set as death from any cause. Secondary end-points were any cardiac events, including non-fatal acute myocardial infarction, coronary artery bypass surgery and repeat PTCA. The mean follow-up period was 4.5 +/- 1.8 years. A multivariate analysis was performed with the Cox proportional-hazard model. The Kaplan-Meier analysis showed that the calcium antagonist (-) group had significantly worse prognoses than the calcium antagonist (+) group (p < 0.05), and that there was no significant difference among the prognoses of the four calcium antagonists groups. The multivariate analysis revealed that the use of a calcium antagonist was one of the independent factors positively contributing to the prognosis. The use of any type of calcium antagonist did not increase mortality in patients who underwent successful elective PTCA, rather, it contributed to a favorable outcome. PMID- 10370395 TI - Negative U-wave as a predictor of antihypertensive treatment effect on regression of echocardiographic hypertrophy in hypertensive patients. AB - To clarify the mechanism and the predictors of the reduction in left ventricular mass (LVM) induced by antihypertensive drugs, forty hypertensive patients were classified according to the presence of left ventricular hypertrophy (LVH) as a defined by echocardiographic LVM findings (LVH group: 27 patients, non-LVH group: 13 patients) and according to the presence of negative U-waves (NU) (NU group: 10 patients, non-NU group: 30 patients). Negative U-waves appeared in the LVH group only (10 of 27 patients). The hemodynamic determinants were investigated as a mechanism of LVM reduction in 38 of these patients who were treated for 2 years with antihypertensive drugs. In the LVH group, thickness of interventricular septum (IVST), posterior wall thickness (PWT) and LVM increased significantly compared to the non-LVH group. In the NU group, the left ventricular end diastolic dimension (LVDD) was significantly larger and the relative wall thickness was significantly smaller compared to the non-NU group in the LVH group, with no difference in LVM between the two groups. Negative U-waves disappeared in all cases after treatment. Significant decreases in LVDD and LV wall thickness were observed in the NU group and significant decrease in LV wall thickness in the non-NU group. LVM index was reduced by 24.0% in the NU group and 9.5% in the non-NU group. The disappearance of negative U-waves was an independent predictor of the reduction of LVH. PMID- 10370396 TI - Diurnal variation of ECG intervals and R or T amplitudes in healthy male subjects assessed by means of spectral and cosinor analysis. AB - Twelve lead standard ECGs 35 healthy male volunteers, recorded from 08.00 H up to 01.00 H, were analysed retrospectively in order to investigate primarily the diurnal changes in vital sign measurements, ECG interval durations (PQ, QRS, QTc) and R wave as well as T wave amplitudes. Data were analysed descriptively and by means of ANOVA, spectral analysis and cosinor models. ANOVA revealed significant variations for vital sign parameters and the QTc interval, although remarkable variation in all ECG parameters was observed. T wave amplitudes in particular showed marked alterations, with changes up to 40%. Transient maxima of median T wave amplitudes and QTc interval duration were found at 12.00 H and 14.00 H, respectively. R wave amplitudes and PQ duration revealed lower values in the afternoon. Although there was remarkable interindividual variability, ultradian rhythmic activity of ECG parameters was confirmed by spectral analysis and cosinor models based on population data. PMID- 10370397 TI - Different effects of temocapril and cadralazine on electrocardiographic voltages and left ventricular mass in patients with essential hypertension. AB - To assess whether electrocardiographic variables are useful to detect the regression of left ventricular (LV) mass after long-term antihypertensive treatment, we related electrocardiographic voltages to echocardiographic variables before and after treatment with an ACE inhibitor, temocapril (TEM), or direct vasodilator, cadralazine (CAD). Twenty-one patients with essential hypertension were treated with either TEM (n = 11) or CAD (n = 10) for one year. LV mass index (LVMI) by echocardiography and Sokolow-Lyon voltage (SV1 + RV5), Cornell voltage (RaVL + SV3) and RV5 + RV6 by standard 12-lead electrocardiographic voltages were determined before and after treatment. Both drugs decreased blood pressure to the same extent. Both Sokolow-Lyon voltage and RV5 + RV6 tended to decrease in the ACE group (40.0 +/- 9.4 to 37.2 +/- 9.4 mm and 44.7 +/- 13.5 to 41.7 +/- 11.7 mm, respectively, N.S.), but not in the CAD group (38.4 +/- 6.8 to 39.7 +/- 7.7 mm and 42.9 +/- 10.4 to 46.8 +/- 11.2 mm, respectively, N.S.). LVMI decreased in the ACE group (-24 +/- 22 g/m2), whereas it increased in the CAD group (37 +/- 27 g/m2, p < 0.01). Change in LVMI was correlated with the changes in RV5 + RV6 and Sokolow-Lyon voltage (r = 0.73, p < 0.01 and r = 0.70, p < 0.01, respectively), but not with that in Cornell voltage. These results indicated that the changes in voltage criteria of RV5 + RV6 and Sokolow-Lyon are useful to assess the change in LVM after antihypertensive treatment in patients with essential hypertension although voltage variables in electrocardiogram were not sensitive to detect changes in LVMI. PMID- 10370398 TI - Lipoprotein(a) concentrations in healthy subjects in the Dominican Republic. Comparison with Japanese. AB - Previous studies have shown that serum concentrations of lipoprotein(a) [Lp(a)] are markedly different among different ethnic groups. We examined the serum levels of total cholesterol, high density lipoprotein (HDL) cholesterol and Lp(a) in apparently healthy subjects aged 20-69 years in Japan (n = 865) and the Dominican Republic (n = 1,893). Dominicans had significantly lower levels of total cholesterol and HDL cholesterol than Japanese. The distribution of Lp(a) concentrations were markedly skewed towards low levels in both Japanese and Dominicans. However, the mean Lp(a) concentration in Dominicans was approximately 2 times higher than in Japanese (21.7 +/- 23.7 vs 12.3 +/- 15.9 mg/dl, p < 0.001). This is possibly because the majority of Dominicans are of mixed Negroid and European blood. PMID- 10370399 TI - Poor reproducibility of false-positive tilt testing results in healthy volunteers. AB - Positive responses to head-up tilt testing occur in healthy subjects. However, the reproducibility of "false-positive" tilt testing results has not been clarified. To study the reproducibility of "false-positive" responses, we prospectively performed 2 tilt tests separated by 1 to 10 (mean 3.2) weeks in 20 healthy males aged 23 to 40 years (mean 30 years). The baseline tilt test (80 degrees for 30 minutes) ended positive in 4 (20%) subjects on the initial test and 2 (10%) on the second test with only 1 (5%) who had consecutive positive responses. No additional positive responses were noted during the isoproterenol (0.01 microgram/kg/min)-tilt test for 10 minutes. We demonstrated that a false positive response occurred in 5 (25%) of 20 young males who underwent 2 tilt tests, however, only 1 (5%) subject had consecutive positive responses. Poor reproducibility may be characteristic of false-positive responses in head-up tilt testing. PMID- 10370400 TI - The effects of antiplatelet agents in the prevention of ventricular tachyarrhythmias during acute myocardial ischemia in rats. AB - Experiments were performed in rat models to study the effectiveness of various antiplatelet agents in the prevention of ventricular tachyarrhythmias during acute myocardial ischemia. The time to the onset of ST-segment elevation and initiating ventricular arrhythmias, frequency and incidence of ventricular arrhythmias, and mortality rates were observed during acute myocardial ischemia (20 minutes) induced by ligation of the proximal left anterior descending coronary artery (LAD) in anesthetized rats. Four groups were studied: Control group (n = 10, not pretreated); Aspirin pretreated group (n = 10, 300 mg/kg p.o. for 1 wk); Ticlopidine pretreated group (n = 10, 200 mg/kg p.o. for 1 wk); and Abciximab (Platelet glycoprotein IIb/IIIa receptor antagonist) pretreated group (n = 10, 2 mg/kg i.v. 10-20 minutes before an experiment). No significant difference was observed in the time to the onset of ST-segment elevation and ventricular arrhythmias between the groups. The incidence of ventricular tachycardia (VT) in the abciximab group was significantly lower than in the control group (p < 0.05) and ventricular fibrillation (VF) in the aspirin and ticlopidine group was significantly lower than in the control group (p < 0.05). The mortality rate in the ticlopidine group was significantly lower than in the control group (p < 0.01). This study suggests aspirin, ticlopidine, and abciximab can effectively prevent VT or VF during acute myocardial ischemia induced by nonthrombotic occlusion and its antiarrhythmic effect may lead to prolonged survival. PMID- 10370401 TI - Ventricular fibrillation with small amplitude of activation and its implications for implantable cardioverter defibrillator treatment. AB - An implantable cardioverter defibrillator (ICD) was implanted in a patient with ventricular fibrillation (VF) related to old myocardial infarction. During VF, amplitude of ventricular activation was small, and the ventricular sensitivity at 1.2 mV failed to detect several small ventricular activations. When the sensitivity was changed to 0.3 mV, both under- and oversensed beats occurred during VF, and at the ventricular sensitivity of 0.15 mV, the undersensed beats disappeared while oversensed beats markedly increased. Defibrillation test was repeated one and four weeks after the implantation, and these inappropriate beats were minimized at the ventricular sensitivity of 0.3 mV. We should pay attention to the amplitude of ventricular activation to avoid possible trouble in ICD therapy. PMID- 10370402 TI - Hypoglycemia induced by interaction between clarithromycin and disopyramide. AB - A 59-year-old man receiving hemodialysis was hospitalized due to severe hypoglycemic attack. The patient had been treated with disopyramide (50 mg/day) because of paroxysmal atrial fibrillation. Hypoglycemia occurred after taking clarithromycin (CAM, 600 mg/day), a macrolide antibiotic. The serum disopyramide concentration reached 8.0 micrograms/ml (23.6 microM) in the presence of CAM, while it was 1.5 micrograms/ml before the addition of CAM. A 75 g oral glucose tolerance test and daily profiles of blood glucose value showed that blood glucose levels were significantly lower in the presence of CAM and disopyramide compared to that in the absence of these drugs. The Turner index in the presence of CAM and disopyramide was significantly higher than that in the absence of these drugs, suggesting that a toxic concentration of disopyramide enhanced insulin secretion, resulting in the induction of hypoglycemic attacks, in which the inhibitory effects of CAM on the hepatic chytochrome P-450 might be involved. QT and QTc intervals were prolonged in the presence of CAM and disopyramide, but torsades de points were not observed in this patient receiving nicorandil (15 mg/day). Thus, it should be taken into account that life-threatening hypoglycemia may result from the interaction between clarithromycin and disopyramide. PMID- 10370403 TI - [The prospects and problems of electron-beam CT (EBT)]. PMID- 10370404 TI - [The multimodal integration, correlation and fusion of morphology and function: the methods and initial clinical applications]. AB - Besides the pure visual diagnosis of morphology in comparison with function, digital multimodal image analysis is steadily gaining in importance. Digital image processing is used in integration, correlation and fusion of topographically identical image data. After defining these terms and describing the acquisition techniques and influence parameters, this article reviews the methods of multimodal image processing, with emphasis on correlation methods. It also gives a short description of special methods in nuclear medicine. The clinical part briefly reviews the clinical use as well as the progress achieved and the benefits expected for diagnostic applications. PMID- 10370405 TI - [Dynamic 31-phosphorus magnetic resonance spectroscopy of the m. quadriceps: the effect of sex and age on the spectroscopic parameters]. AB - PURPOSE: 31P-MRS is used to assess the influence of sex und age on quadriceps muscle metabolism before and after exercise. MATERIALS AND METHODS: 32 healthy volunteers (15 women, 17 men, mean age: 38 +/- 17 yrs.) were examined by dynamic phosphorus-31 (31P) magnetic resonance spectroscopy (MRS). In the magnet, the quadriceps muscle was stressed by an isometric und an isotonic form of exercise until exhaustion, respectively. RESULTS: Resting conditions: With increasing subjects' age, the ratio beta-adenosine triphosphate/total phosphate decreased (r = -0.37; p = 0.02). With increasing subjects' age, the ratios inorganic phosphate/phosphocreatine (r = 0.79; p = 5 x 10(-8), phosphomonoester/beta adenosine triphosphate (r = 0.74; p = 10(-6) and phosphodiester/beta-adenosine triphosphate (r = 0.62; p = 10(-4) increased. The pH was the only one of the evaluated spectroscopic parameters which showed a sex-dependence: Female subjects had a significantly lower pH (7.03 +/- 0.02) than male subjects (7.05 +/- 0.03; p = 0.01). Exercise: With increasing age, the maxima of inorganic phosphate/phosphocreatine were less extreme during both of the exercises (r = 0.42; p = 0.0005). Likewise, the exercise-induced acidosis was less severe with increasing age (r = 0.53; p = 6 x 10(-6). After the end of the exercise, the times of half recovery of inorganic phosphate/phosphocreatine and the pH correlated neither with the subjects' age nor with sex or cross-sectional areas of the quadriceps muscle. CONCLUSION: Sex and age of volunteers affect spectroscopic results. This influence had to be considered in the interpretation of spectroscopic studies. PMID- 10370406 TI - [The determination of left and right ventricular ejection and filling functions of the heart by fast cine MRT in the breath-hold technic in subjects]. AB - PURPOSE: Evaluation and comparison of localized and global left and right ventricular ejection and filling with fast cine MR imaging in the breath-hold technique. MATERIALS AND METHODS: 10 healthy volunteers were examined with a 1.5 Tesla unit and phased-array-coil using a segmented FLASH-2D sequence in breath hold technique. Peak ejection and peak filling rates [PER, PFR end-diastolic volume (EDV)/s)]. time to PER and PFR [TPER, TPFR ms] and time of end-systole [TSYS in % RR-interval] of all slices (complete-slice-evaluation) were evaluated and compared to three left ventricular and one right ventricular slices (reduced three-slice-evaluation). RESULTS: There were significant regional left ventricular differences of PER (p = 0.002) and PFR (p = 0.007), but not of TPER and TPFR. Ejection and filling indices of the left ventricular middle slice were closest to the overall evaluation of all sections. In the left-/right-side comparison the right ventricular PFR was higher than the left ventricular (5.1 and 4.2 EDV/s) and the right ventricular TPFR was earlier than the left (92.2 and 123.5 ms). CONCLUSIONS: With fast cine techniques, regional and global left and right ventricular ejection and filling indices can be evaluated in addition to the global heart volume indices. The three-slice-evaluation represents a comprehensive, clear and time-saving method for daily routine. PMID- 10370408 TI - [MRT versus CT in the diagnosis of pneumonias: an evaluation of a T2-weighted ultrafast turbo-spin-echo sequence (UTSE)]. AB - PURPOSE: To evaluate a T2-weighted URSE sequence for the assessment of pulmonary infiltrations in comparison to CT. METHODS: 28 MRT scans of 22 patients with confirmed pneumonia were recorded on a 1.5 Tesla apparatus with an expiratory and diastolic triggered, T2-weighted ultrafast-spin-echo sequence in axial slice mode with the following parameters: TReff/TE/Turbo-factor 2000-4000/90 ms/21-23; slice thickness/separation 6/0.6 mm; FOV 360 mm; 24 slices. 24 spiral CTs (since thickness/table advance: 1-2 mm/10mm) were available for comparison. The separate evaluation of MRTs and CTs was performed by three radiologists in a consensus procedure with regard to pulmonary lesions (e.g., infiltration, round foci, net patterns) and image quality of the MRTs (4-step scale). RESULTS: In 71% of the cases the CTs and MRTs agreed with the diagnosis and representation of the lesions, in 25% MRT was superior. MRT was better for the detection of pulmonary abscesses. In 93% the image quality of the MRT was very good to good. CONCLUSIONS: MRT in the technique presented here is in most cases equal to CT for the detection of pneumonia. Diagnosis of pulmonary abscesses seems to be better with MRT. PMID- 10370407 TI - [The assessment of the patency of coronary bypass vessels with a 2D T2-weighted turbo-spin-echo sequence (HASTE) in the breath-hold technic]. AB - AIM: To evaluate the patency of coronary artery bypass grafts with a 2D T2 weighted breath-hold turbo-spin-echo sequence. METHODS: 38 patients with 97 grafts (19 internal mammary artery and 78 saphenous vein grafts) and a total of 120 distal anastomoses were studied at 1.5 Tesla in supine position using a phased array body coil. An ECG gated 2D T2-weighted breath-hold turbo-spin-echo sequence (HASTE) was performed. Reference method was selective coronary angiography. The image material was evaluated independently by two radiologists (observer one, a radiological fellow and the second a staff radiologist). RESULTS: Observer 1 reached a sensitivity of 96% (72/75) and a specificity of 91% (20/22), positive predictive value was 97%, negative predictive value 87%. 79 of the 97 (81%) patent distal anastomoses were correctly identified. Observer 2 achieved a sensitivity of 92% (69/75) and a specificity of 82% (18/22), positive and negative predictive values were 95% and 75% respectively. From 97 patent distal anastomoses 59 (61%) were recognized. The interobserver agreement was good (Cohen's kappa = 68%, p-value [McNemar] = 58%). CONCLUSION: Using the HASTE sequence a reliable assessment of graft patency is possible. This sequence is a helpful tool for planning flow measurements and 3D MR angiography. PMID- 10370409 TI - [The extracapsular spread of cervical lymph node metastases: the diagnostic value of computed tomography]. AB - OBJECTIVE: To assess the value of computed tomography in the determination of extracapsular neoplastic spread and soft tissue infiltration of cervical lymph nodes. MATERIALS AND METHODS: Prospectively 165 CT reports of patients with squamous cell carcinoma in the head-neck region were compared with the histologic specimens after neck dissection. RESULTS: CT reached a sensitivity of 80.9% in the determination of extra-capsular neoplastic spread. The infiltration of cervical muscles and the jugular vein was always determined but-often diagnosed false-positive. Therefore, the overall specificity in determining extracapsular neoplastic spread was low with 72.7%. CONCLUSIONS: The accuracy of computed tomography (76.3%) in the determination of extracapsular neoplastic spread, infiltration of cervical muscles and the jugular vein is only partially satisfying. PMID- 10370410 TI - [The value of magnetic resonance tomography (MRT), magnetic resonance cholangiopancreatography (MRCP) and endoscopic retrograde cholangiopancreatography (ERCP) in the diagnosis of pancreatic tumors]. AB - PURPOSE: To prospectively evaluate the role of MRI including MR cholangiopancreatography (MRCP) compared to endoscopic retrograde cholangiopancreatography (ERCP) in the diagnosis of pancreatic cancer. MATERIAL AND METHODS: ERCP and MRI including MRCP were performed in 52 patients with suspected pancreatic cancer. MRCP was obtained using a single-shot RARE technique. The results of axial images and MRCP were compared to concurrently performed ERCP examinations. The standards of reference were the surgical and pathological findings, respectively. Image quality of MRCP was assessed using a three-step-score (1 = good, 2 = fair, 3 = nondiagnostic). RESULTS: In 88% of the cases the MRCP was of good quality. Only in 4% was MRCP non-diagnostic. The combination of MRI and MRCP showed an overall accuracy of 88%, whereas the overall accuracies of MRCP alone and ERCP were 80%, and 85%, respectively. The positive predictive values of MRI/MRCP, MRCP alone, and ERCP were 91%, 85%, and 88%, respectively. CONCLUSION: For the detection of pancreatic cancer MRI including MRCP is comparable to ERCP and can be regarded as the method of choice in patients with suspected pancreatic cancer. ERCP is the procedure of choice in patients with contraindications to MRI and in patients in whom additional therapeutic procedures are performed. PMID- 10370411 TI - [3D endosonography to clarify distal ureteral processes]. AB - PURPOSE: Evaluation of 3D endoluminal sonography as a diagnostic modality in lower ureteral pathologies. MATERIALS AND METHODS: Between December 93 and December 97, 36 consecutive patients with negative findings on IVP and still suspected ureteral obstruction were referred for transrectal or transvaginal endosonography. Patients mean age was 63 years and all of them presented clinically obvious symptoms such as colic pain and miction disturbances. RESULTS: In 31 of 36 patients (86%) 3D endosonography was diagnostic although previous IVP was negative. Reasons for ureteral obstructions were ureteral calculi in 23, urological tumours in 5 and rectal carcinoma in two cases. In one patients a gynecological tumour caused the symptomatology. In the remaining 5 patients CT/MRT or invasive retrograde ureterography/ureteroscopy had to be performed for the final diagnosis. CONCLUSIONS: The use of 3D endosonography shows encouraging results in the diagnosis of distal ureteral pathologies. In case of negative findings on IVP it should therefore precede invasive diagnostic modalities or cost intensive imaging techniques. PMID- 10370412 TI - [The characteristic values in the MR study of cerebral blood flow with high spatial and temporal resolution]. AB - PURPOSE: Different parameters for the evaluation of perfusion studies with dynamic MR measurements after the administration of paramagnetic contrast agent were compared. METHOD: An echo planar imaging (EPI) sequence was developed that allows dynamic multi-slice data acquisition. Evaluations of the measurements were performed by calculating parameter maps. Phenomenological quantities such as maximal signal reduction and time to peak as well as derived quantities such as approximations of the regional cerebral blood volume were used as parameters. Patients with a low grade astrocytoma, with an acute cerebral infarction and with stenoses of the carotid arteries were examined. RESULTS: On comparison of the derived parameters with the phenomenological parameters no major differences were found in qualitative comparisons of the parameter maps. The comparison of relative parameter values in selected regions within the datasets of 20 patients show a high correspondence between phenomenological and derived parameter values. CONCLUSIONS: Using EPI measurement techniques with high temporal resolution, parameter maps of simple values like maximal signal reduction and time-to-peak are sufficient to visualize relative differences of perfusion within the selected slices. However, the calculation of relative rCBV and rCBF values might be helpful for the detailed characterization of signal courses in selected regions. PMID- 10370413 TI - [Digital 3D rotational angiography for the preoperative and preinterventional clarification of cerebral arterial aneurysms]. AB - OBJECTIVE: Does the recently introduced 3D angiography provide additional information beyond standard angiography (DSA) for the diagnosis of cerebral aneurysms? METHODS: During a 3-months period DSA and 3D-angiography were performed in 40 patients harbouring a total of 49 aneurysms. Vascular regions that presented an aneurysm diagnosed by DSA were reevaluated by 3D-angiography. RESULTS: In two patients, vessel-loops previously described as aneurysms by DSA could be identified by 3D-angiography. In one patient, an aneurysm was diagnosed that could not be detected by DSA. In another case, the definitive diagnosis of an aneurysm was obtained only with 3D-angiography. In one patient, an aneurysm was diagnosed that could not be detected by DSA. In another case, the diagnosis of an aneurysm was obtained only with 3D-angiography. In two cases, aneurysms could be definitively excluded by 3D-angiography, whereas in another aneurysm a vessel originating from this lesion was identified. The size of the aneurysms measured by both methods was identical. CONCLUSIONS: Multiple projections of 3D angiography provide a better evaluation of the anatomic situation regarding the base of the aneurysm as well as the relationship of an aneurysm to neighbouring vessels. Further, an exact differentiation between a vessel loop and an aneurysm can be made. Therefore, 3D-angiography is a valuable tool when used in conjunction with DSA. PMID- 10370414 TI - [3D-TSE MR cholangiopancreatography with breath triggering to clarify unclear hepatopathies in children]. AB - PURPOSE: Evaluation of 3D-TSE MR-cholangiography with respiratory triggering in the work up of hepatopathies in infants and young children. PATIENTS AND METHOD: 16 infants (4-16 years) with increased transaminases, two with recurrent pancreatitis, were examined at 1.5 T (ACS-NT II, Philips Medical Systems) using a 3D-TSE MRCP with respiratory triggering in addition to a regular MRI of the liver. The MRCP was compared to ERCP. Two radiologists and one gastroenterologist evaluated the technical quality, visualization of the pancreaticobiliary system, and the diagnostic value of the examinations. RESULTS: Technically feasible were 14/16 MRCPs and 13/16 ERCPs. Two MRCP were not of diagnostic value due to motion artifacts and in three ERCP cannulation of the papilla was not possible. 14/16 ERCP required general anaesthesia, while MRCP needed i.v. sedation in two patients only. Extrahepatic ducts/cystic duct/pancreatic duct were visualized in 14/12/8 patients using MRCP, and in 13/10/3 patients using ERCP, both without adverse effects or complications. Intrahepatic ducts were better delineated with MRCP. In 10 patients with histologically proven periportal fibrosis (n = 7) and liver fibrosis (n = 1) or antineutrophil cytoplasmatic antibodies and associated inflammatory bowel disease, MRCP and ERCP revealed pathological results. CONCLUSION: MRCP using a 3D-TSE sequence with respiratory triggering is a good non-invasive technique for delineation of the biliary tract in infants and young children for the work up to hepatopathies. PMID- 10370415 TI - [The Perflex stent, a new balloon-expandable vascular stent: the initial clinical results]. AB - PURPOSE: Clinical evaluation of a newly developed balloon-expandable vascular stent. METHOD: Participating in a multi-center study our clinic enrolled 12 patients with 15 iliac artery stenoses (Fontaine IIa-III) that were treated with the new stent. Three patients had bilateral stenoses. The mean ankle-brachial index (ABI) at rest was 0.76 +/- 0.25 (+/- 1 standard deviation), the mean ABI after exercise was 0.47 +/- 0.20. Indication for stent insertion was a residual trans-lesion pressure gradient > or = 10 mmHg after balloon angioplasty. Follow up included clinical examination, measurements of the ABI at rest and after exercise, and colour duplex ultrasound (CDS) on the day of hospital dismissal and 1, 6 and 12 months after stent insertion. RESULTS: 16 stents were placed successfully. One stenosis was treated with two stents. Acute complications did not occur. The Fontaine stage increased by at least one stage in all patients. The ABI at rest rose to 0.91 +/- 0.26, the ABI after exercise to 0.86 +/- 0.29. During follow-up one stent occlusion, one stenosis proximal to a stent, and CDS findings suggesting restenosis in two further stents were observed. CONCLUSIONS: It appears that the Perflex stent may be used in the iliac arteries with a success rate similar to other stents. However, this preliminary result has to be confirmed in a larger series and after a longer follow-up. PMID- 10370416 TI - [Direct digital magnification mammography with a large-surface detector made of amorphous silicon]. AB - PURPOSE: To investigate the image quality of a new direct digital mammography system using a large-area amorphous silicon X-ray detector in a phantom study. MATERIALS AND METHODS: The contrast-detail resolution as a function of the tube voltages, the magnification factors and the mean glandular doses were investigated using dedicated test objects. RESULTS: The contrast-detail resolution was significantly improved in comparison with conventional screen-film mammography. Usually, the doses necessary to obtain these high-quality survey mammograms were smaller. CONCLUSIONS: By combining the direct magnification technique and a digital flat panel detector the limited spatial resolution of such image receptors can be overcome. With this direct digital mammography technique, a digital image was directly captured without an intermediate step of optical or mechanical scan. PMID- 10370417 TI - [Primary bone tumors and "tumor-like lesions" of the shoulder. Their histopathology and imaging]. AB - Purpose of this review is to demonstrate typical X-ray, CT and MR morphology of primary bone tumors and "tumor-like lesions" of the shoulder in correlation with histopathology. 711 primary bone tumors of the shoulder and proximal humerus were studied. 602 were localized in the humerus, 90 in the scapula and 19 in the clavicula. The most frequent benign tumors were osteochondromas (n = 143), simple bone cysts (n = 115), enchondromas (n = 75) and aneurysmal bone cysts (n = 50). Fibrous dysplasia (n = 25), chondroblastoma (n = 15), osteoid osteoma (n = 13), giant cell tumors (n = 12) and non ossifying fibroma (n = 11) were less frequent. The most frequent malignant bone tumors were osteosarcoma (n = 72), chondrosarcoma (n = 52) and Ewing's sarcoma (n = 46). Focal plasmocytoma (n = 20) and lymphoma (n = 10) were less frequent. The average age of the patients was 31.5 years. Some of these tumors were typically located in the shoulder, i.e. simple bone cysts and chondroblastoma. In summary the shoulder was a rather infrequent site of primary bone tumors, but since most of these tumors were benign, the radiologist should be aware of the differential diagnosis to guide therapy. PMID- 10370418 TI - [The complete disintegration of microcalcification in a ductal invasive breast carcinoma]. PMID- 10370419 TI - [Gout in the area of the cervical vertebrae and the sternoclavicular joint]. PMID- 10370420 TI - [Abscessing cavernitis--its imaging by MRT]. PMID- 10370421 TI - [Ganglioglioma of the cerebellum on the CT]. PMID- 10370428 TI - Deinstitutionalization at the beginning of the new millennium. AB - Deinstitutionalization, which began in the mid-1950s, has had dramatic effects. It has decreased the number of occupied state hospital beds from 339 to 29 per 100,000 population. The plight of the new generation of chronically and severely mentally ill persons has posed the most serious problem: these individuals no longer receive life-long hospital admission and thus permanent asylum from the demands of the world. A comprehensive, integrated, and adequately funded system of care for this population needs to be established in the community. Where such systems exist, they can lead to higher levels of functioning and a better quality of life. Unfortunately, treatment, housing, and rehabilitation resources are presently insufficient to serve the substantial population of mentally ill in the community. Moreover, some patients who have been deinstitutionalized cannot be effectively treated without highly structured 24-hour care. The way deinstitutionalization has been implemented has probably contributed to the large numbers of severely mentally ill persons on the streets and in jails. Although deinstitutionalization can result in a much richer life experience in the community, much more needs to be done to make that occur. PMID- 10370429 TI - Experimental approaches to cognitive disturbance in Alzheimer's disease. AB - This paper reviews available and potential treatments for the cognitive disturbances associated with Alzheimer's disease. The neurochemical, neuropathological, and molecular-biological abnormalities associated with this disorder, as well as possible sites for pharmacological intervention, are discussed. These sites include genetic alterations in apolipoprotein E, amyloid precursor protein, and presenilin. Additionally, modification of amyloid processing, tau processing, and calcium regulation may have a role in future treatment. Intriguing epidemiological findings involving antiinflammatories, antioxidants, and estrogen for the cognitive deficits associated with Alzheimer's disease suggest the need for clinical trials of these agents. The current status of cholinesterase inhibitors, muscarinic receptor agonists, nicotine, and adrenergic and glutaminergic approaches to treatment are described. PMID- 10370430 TI - Population-based guidelines for performance measurement: a preliminary report. AB - This paper describes the development of--and early efforts to validate- guidelines that indicate average amounts of service expected to be used by a population of patients with a given disorder who are served by a comprehensive mental health system. These guidelines address expected service use by individuals in 55 diagnostic groups. The purpose of these guidelines is to provide a gauge for evaluating the amounts of service being delivered by managed care organizations. Three population-based guidelines (for attention deficit/hyperactivity disorder, major depressive disorder, and schizophrenia) are compared to actual amounts of service delivered to enrollees in large behavioral health care systems. PMID- 10370431 TI - Psychotic acts: the question of meaning. PMID- 10370433 TI - Inpatient residency training in a time of change. PMID- 10370432 TI - The psychotherapist-patient privilege. PMID- 10370434 TI - Generativity and community in mental health: continuity and challenge. PMID- 10370435 TI - Clozapine for comorbid substance use disorder and schizophrenia: do patients with schizophrenia have a reward-deficiency syndrome that can be ameliorated by clozapine? AB - Alcohol and other drugs of abuse are commonly used by persons with schizophrenia and contribute to the overall morbidity of the disorder. Standard, or typical, antipsychotic drugs do not limit such substance use and may even render it more likely. However, preliminary data from our group and others suggest that the atypical antipsychotic clozapine may decrease substance use in this population. While recognizing the likelihood that substance use decreases negative symptoms (as well as extrapyramidal symptoms) in persons with schizophrenia, we hypothesize that the biological basis of substance use relates to a "reward deficiency syndrome" secondary to dysfunctional dopamine-mediated mesocorticolimbic neurons in these individuals. We further suggest that clozapine's beneficial effect in patients with comorbid schizophrenia and substance use disorders may relate to its presumed ability to ameliorate the deficits in both the mesocortical and mesolimbic dopaminergic neuronal projections through its various actions on dopaminergic, serotonergic, and particularly noradrenergic neurons. PMID- 10370436 TI - Sex differences in marijuana use in the United States. AB - Marijuana and other cannabis preparations are the most widely used illicit drugs in the United States. A review of the literature reveals a number of sex differences in the epidemiology and adverse medical consequences of marijuana use. In 1995, 6.5% of females and 10.5% of males aged 12 and older reported marijuana use in the previous year. Although 4% more males than females used marijuana, the percentage of males using marijuana between 1994 and 1995 had decreased, whereas the percentage of females using marijuana during that same period had increased. Among females, the age of initiation of use is declining and the prevalence of problems with marijuana is on the rise. Both male and female marijuana users may experience adverse effects of cognitive dysfunction and airway inflammation. However, clinicians should be aware of sex-specific effects of marijuana use, including a possible increased risk of prostate cancer for male users and possible adverse effects on reproductive hormones in female users. Review of the available information on this topic indicates that we have much more to learn about the similarities and differences between males and females with respect to patterns of use, adverse consequences, and vulnerabilities to marijuana. PMID- 10370437 TI - Chronic posttraumatic stress disorder: a review and checklist of factors influencing prognosis. AB - Mental health clinicians are often asked to evaluate prognosis in individuals with posttraumatic stress disorder (PTSD) in clinical, administrative, and legal contexts. Although chronicity of PTSD has been addressed in a number of trauma studies, the data have not been integrated into a coherent approach to the assessment of prognosis. In this paper, the peer-reviewed PTSD literature is surveyed to assist clinicians in making informed prognostic evaluations of the course of PTSD in adults. Potential risk factors, grouped into 11 categories (PTSD stressors, PTSD symptoms, current comorbidity, lifetime comorbidity, childhood separation and abuse, demographics, life stressors, family history, support, treatment, and functional impairment), are reviewed. Knowledge of these risk factors, and of factors associated with chronic PTSD, is helpful in assessing the potential for or degree of chronicity present at the initial evaluation of the patient, as well as in measuring treatment response during the course of therapy. Early identification and the appropriate treatment and management of remediable risk and associated factors may help prevent the development of chronic PTSD. Longitudinally assessing the response of treatable risk factors should provide an additional means for evaluating prognosis. A PTSD Prognostic Checklist, which rates risk and associated factors in each category, is proposed. Validity and reliability have not yet been established for this instrument. It is hoped that clinicians will use and conduct research on it as an initial step toward advancing its scientific utility. PMID- 10370438 TI - Autistic symptoms in a 21-year-old college student: perspectives on diagnosis and treatment. PMID- 10370439 TI - Strategies for augmentation of SSRI treatment: a survey of an academic psychopharmacology practice. PMID- 10370440 TI - The development of National Alcohol Screening Day. PMID- 10370441 TI - Ice-nine and human prion disease. PMID- 10370442 TI - Replacing cost with value. PMID- 10370443 TI - Antidepressant-induced sexual dysfunction: review, classification, and suggestions for treatment. AB - Sexual function, an important component of quality of life, is often affected by antidepressant treatment. Reports associate antidepressant medications with a wide range of sexual disorders of desire, arousal, and orgasm, and with the occurrence of sexual pain. Fewer sexual dysfunctions have been reported with bupropion, nefazodone, and mirtazapine than with the monoamine-oxidase inhibitors, tricyclic antidepressants, selective serotonin-reuptake inhibitors, and venlafaxine. Sexual dysfunctions may occur in more than half of patients treated with selective serotonin-reuptake inhibitors, but patients may not readily divulge sexual information unless a clinician is knowledgeable and proactive in assessment. Once an antidepressant-induced sexual dysfunction is detected and its nature is characterized, an appropriate treatment intervention can be chosen in order to alleviate the sexual disorder and enhance treatment compliance. This review classifies antidepressant-induced sexual dysfunctions, discusses assessment and differential diagnosis, and describes currently reported treatment approaches. PMID- 10370444 TI - Primary and drug-induced disorders of water homeostasis in psychiatric patients: principles of diagnosis and management. AB - Psychotropic drugs, as well as some psychiatric disorders, can produce neurotoxic and life-threatening abnormalities of water and electrolyte balance that require prompt and appropriate medical intervention. Compulsive fluid intake by psychotic patients (primary polydipsia) can produce delirium due to water intoxication with hyponatremia. Several psychotropic drugs cause water retention by decreasing renal clearance, as in the syndrome of inappropriate antidiuretic hormone secretion. Lithium and other agents interfere with renal resorption of water to cause nephrogenic diabetes insipidus. Clinical signs in these disorders range from lethargy and confusion to stupor, seizures, coma, and death. This overview provides a conceptual framework for differentiating among and safely managing these relatively common disorders. PMID- 10370445 TI - The pain of being borderline: dysphoric states specific to borderline personality disorder. AB - The objective of this study was to identify the dysphoric states that best characterize patients meeting criteria for borderline personality disorder and distinguish them from those in patients with other forms of personality disorder. One hundred forty-six patients with criteria-defined borderline personality disorder and 34 Axis II controls filled out the Dysphoric Affect Scale, a 50-item self-report measure that was designed for this purpose and has good internal consistency and test-retest reliability. Twenty-five dysphoric states (mostly affects) were found to be significantly more common among borderline patients than controls but nonspecific to borderline personality disorder. Twenty-five other dysphoric states (mostly cognitions) were found to be both significantly more common among borderline patients than controls and highly specific to borderline personality disorder. These states tended to fall into one of four clusters: (1) extreme feelings, (2) destructiveness or self-destructiveness, (3) fragmentation or "identitylessness," and (4) victimization. In addition, three of the 25 more-specific states (feeling betrayed, like hurting myself, and completely out of control), when occurring together, were particularly strongly associated with the borderline diagnosis. Equally important, overall mean Dysphoric Affect Scale scores correctly distinguished borderline personality disorder from other personality disorders in 84% of the subjects. Taken together, the results of this study suggest that the subjective pain of borderline patients may be both more pervasive and more multifaceted than previously recognized, and that the overall "amplitude" of this pain may be a particularly good marker for the borderline diagnosis. PMID- 10370446 TI - Identity and intimacy in psychotherapy with a young woman: four perspectives. PMID- 10370447 TI - Deinstitutionalization and community care: social welfare policy as mental health policy. PMID- 10370448 TI - The courage to sit: one resident's reflections on the psychiatry training process. PMID- 10370449 TI - Treatment algorithms: bane or boon to mental health? PMID- 10370450 TI - Negative symptoms in schizophrenia: neurobiological models and treatment response. AB - Evidence from lesioning studies and neuroimaging has linked negative symptoms to dysfunction of the prefrontal cortex, the limbic system, and the basal ganglia. Although such symptoms have been most strongly associated with dopaminergic hypoactivity in the prefrontal cortex, other neurotransmitters including norepinephrine, serotonin, and the excitatory amino acids may also play a role. In some patients moderate doses of conventional neuroleptics clearly improve negative symptoms; the response of such symptoms is relatively greater with clozapine and probably with certain serotonin-dopamine antagonists. Recent studies demonstrating improvement of negative symptoms when conventional neuroleptics are augmented with selective serotonin-reuptake inhibitors or with agents active at the glycine-modulatory site of the glutamatergic N-methyl-D aspartate receptor complex suggest that further amelioration of primary negative symptoms may be possible through pharmacological strategies involving multiple neurotransmitter systems. PMID- 10370451 TI - Talisman or taboo: the controversy of the suicide-prevention contract. AB - The suicide-prevention contract is a widely used but overvalued clinical and risk management technique. The scant information on this topic in the psychiatric and mental health literature is reviewed, along with the literature on collateral subjects including suicide prediction, medicolegal aspects of treating suicidal patients, the therapeutic alliance, and countertransference with suicidal patients. A group of 112 psychiatrists and psychologists was surveyed about their use of suicide-prevention contracts; the majority of them had never received any formal training on the topic. A combination of factors--the unpredictability of suicide, the many different antecedents to completed suicides, the complex psychological reactions of clinicians (including fear of litigation), the incongruity between clinical and legal usages of the contract concept, and the hazards that come of collapsing a complex treatment process into a few words- limit the applicability of suicide-prevention contracts. We reason that the use of these contracts is based upon subjective belief rather than on objective data or formal training. We recommend an alternative approach to suicide risk management rooted in the well-known and well-defined principles of informed consent. PMID- 10370453 TI - Two views of a delusion. PMID- 10370452 TI - Assessing the quality of psychiatric care: research methods and application in clinical practice. AB - The drive to contain the costs of health care in the United States is focusing attention on how quality of care is affected. This article discusses research methods for assessing the quality of psychiatric care and reviews findings from some major studies evaluating care. These findings are mixed, highlighting areas in which quality of care is less than optimal, as well as the importance of continued research and the need to develop better research methods. Evidence based criteria and more-sensitive risk-adjustment techniques must be employed if data on quality are to yield fair comparisons among health plans. The challenge is to refine the methods now in use at both the research and clinical levels, so that better-quality assessments can be made for policy formulation, physician education, and consumer choice. PMID- 10370454 TI - Treatment of depression during pregnancy: balancing the risks. PMID- 10370455 TI - On spending other people's money. PMID- 10370456 TI - Looking a gift horse in the mouth: the ethics of gift-giving in psychiatry. PMID- 10370457 TI - [Antibiotic therapy at home, an alternative of choice to hospitalization]. PMID- 10370458 TI - [Terminal and palliative care at home]. PMID- 10370459 TI - [Clinical research in pediatric practice applied to ambulatory infectious pathology]. PMID- 10370460 TI - [Home enteral and parenteral nutrition]. PMID- 10370461 TI - [Nutritional needs in the normal adolescent]. PMID- 10370462 TI - [Nutrition in the normal adolescent, iron requirement: effective nutritional requirement]. PMID- 10370463 TI - [The property of calcium in the child and the adolescent: importance in the acquisition of bone mineral density]. PMID- 10370464 TI - [Genetics of overweight and obesity]. PMID- 10370465 TI - [Inhaled medications: the application of nebulization in France]. PMID- 10370466 TI - [Aerosols are a game for children]. PMID- 10370467 TI - [Corticoids and nebulization]. PMID- 10370468 TI - [Antibiotics and nebulization]. PMID- 10370469 TI - [Aerosols in pediatrics: present and future]. PMID- 10370470 TI - [When should one treat a child with chronic hepatitis C with interferon?]. PMID- 10370471 TI - [Natural history of hepatitis C in children]. PMID- 10370472 TI - [Should one treat children with chronic hepatitis C with interferon?]. PMID- 10370473 TI - [New therapeutic possibilities in the treatment of hepatitis B]. PMID- 10370474 TI - [Serious hepatitis A in a child]. PMID- 10370475 TI - [Viral hepatitis: vaccination for hepatitis A and B virus]. PMID- 10370476 TI - [Factors in brain growth and development]. PMID- 10370477 TI - [Development of cortico-cortical connections in primates]. PMID- 10370478 TI - [Genetics and neurobiology of development]. PMID- 10370479 TI - [Monoaminergic neurotransmission and brain dysfunctions in the newborn]. PMID- 10370480 TI - [Functional imagery of cerebral anoxia-ischemia in the newborn]. PMID- 10370481 TI - [Disturbances in aminoglycan synthesis]. PMID- 10370482 TI - [Carbohydrate-deficient glycoprotein syndrome and glycosylation of N glycoproteins]. PMID- 10370484 TI - [Pediatric experiences of a perinatal network organization]. PMID- 10370483 TI - [The Burgundy neonatal program]. PMID- 10370485 TI - [Practical problems of the functioning of the perinatal network in the Loire region]. PMID- 10370486 TI - [Perinatal networks--operation in the Centre region]. PMID- 10370487 TI - [Experiences of the division of neonatology of Lausanne]. PMID- 10370488 TI - [Interest in the study of dose-response relationship for determination of the optimal dose of medications]. PMID- 10370489 TI - [Pharmacology of antipyretics: applications to their use in pediatrics]. PMID- 10370490 TI - [Concern of pharmacovigilance: proarrhythmic effects of medications in current use]. PMID- 10370491 TI - [Pediatric consequences of prenatal diagnosis of heart abnormalities and rhythm problems]. PMID- 10370492 TI - [Diaphragmatic hernias]. PMID- 10370493 TI - [Toxoplasmosis]. PMID- 10370494 TI - [Pediatric consequences of prenatal diagnosis: uropathies]. PMID- 10370495 TI - [Prenatal diagnosis of fetal abnormalities: importance of a prognostic evaluation]. PMID- 10370496 TI - [Contribution of genetics]. PMID- 10370497 TI - [Notification of abnormalities and handicaps before and after birth]. PMID- 10370498 TI - [Ethical aspects of prenatal diagnosis and prognosis]. PMID- 10370499 TI - [Consequences of prenatal diagnosis on the organization of care]. PMID- 10370500 TI - [Conditions and modalities of discharge of the premature newborn]. PMID- 10370501 TI - [Discharge to the home of the large premature infant. Findings of North Pas-de Calais (Epipage). Regional Group of Perinatal Epidemiology]. PMID- 10370502 TI - [Psychosocial problems posed by the discharge to the home of premature newborns]. PMID- 10370503 TI - [A Scandinavian experience]. PMID- 10370504 TI - [Follow-up of the premature infant at home--cooperation between town and home]. PMID- 10370505 TI - [Care of the newborn at Cotonou. Actual state and difficulties]. PMID- 10370506 TI - [Neonatology in the Maghreb region: cost]. PMID- 10370507 TI - [Therapeutic alternatives in neonatology: towards a reduction of the cost of prematurity]. PMID- 10370508 TI - [Childhood asthma and atmospheric pollution]. PMID- 10370509 TI - [Lead poisoning in children]. PMID- 10370510 TI - [Effect of noise on the health of the child]. PMID- 10370511 TI - [Life rhythms in children]. PMID- 10370512 TI - [Drug dermatitis: from benign to serious forms]. PMID- 10370513 TI - [Physiopathology and care of toxic epidermal necrolysis]. PMID- 10370514 TI - [Angiomas and major tissue dysplasias. Pathogenic origins and management]. PMID- 10370515 TI - [Burns: prehospital management]. PMID- 10370517 TI - [Management of heart diseases in children in this country in the process of development]. PMID- 10370516 TI - [ Head and truncal abnormalities and secondary clinical aspects of 22q11 microdeletion. A series of 111 patients]. PMID- 10370518 TI - [Serious illness and prevention of sudden infant death]. PMID- 10370519 TI - [Management of child AIDS victims in an African milieu]. PMID- 10370520 TI - [Prevention of accidents on public roads in children in Lebanon. Is there a path to follow?]. PMID- 10370521 TI - [Acute intestinal invagination: pneumatic reduction (experience with 2033 cases)]. PMID- 10370522 TI - [Cysticercosis]. PMID- 10370523 TI - [Prevention of rotavirus diarrhea by vaccination]. PMID- 10370524 TI - [Important factors in rotavirus infections]. PMID- 10370525 TI - [Vaccines for rotavirus infections: efficacy and eventual role of vaccination programs]. PMID- 10370526 TI - [Cure of childhood cancers]. PMID- 10370527 TI - [Good examples: nephroblastomas, localized neuroblastomas]. PMID- 10370528 TI - [Solid tumors: sarcomas of bone and soft tissue]. PMID- 10370529 TI - [Metastatic tumors, brain tumors]. PMID- 10370530 TI - [Early detection and medical management of sickle cell anemia: five years of experience in Cotonou]. PMID- 10370531 TI - [Hematopoietic stem cell transplantation in sickle cell anemia]. PMID- 10370532 TI - [Vision development in the infant: a plea for testing]. PMID- 10370533 TI - [Evoked otoacoustic emissions in the newborn]. PMID- 10370534 TI - [Evoked otoacoustic emissions at birth: the Luxembourg experience]. PMID- 10370535 TI - [Presentation of apparatus and methods of measurement]. PMID- 10370536 TI - [Technical aid in the management of an asthmatic crisis: measurement of peak expiratory flow rate and transcutaneous pO2]. PMID- 10370537 TI - [Technical help in following the asthmatic child]. PMID- 10370538 TI - [Distorted mother/baby interactions: early symptomatic manifestations]. PMID- 10370539 TI - [Postnatal depression is also of interest in pediatrics]. PMID- 10370540 TI - [Perinatal maternal depression: effects on the baby and young child]. PMID- 10370541 TI - [Difficulty with school work]. PMID- 10370542 TI - [The link between anxiety dependence with school]. PMID- 10370543 TI - [Normal language in language delay]. PMID- 10370544 TI - [Dysphasias of development]. PMID- 10370545 TI - [Oral and written language]. PMID- 10370546 TI - [The psychiatric point of view]. PMID- 10370547 TI - ["Life threatening events in infancy": plea for a semiologic approach and rationalization of examinations]. PMID- 10370548 TI - [Polyvalent immunoglobins in general pediatrics: what place, what risk, and what cost?]. PMID- 10370549 TI - [Pediatrics in terms of civil and professional jurisdiction]. PMID- 10370550 TI - [Antibiotic therapy of acute pyelonephritis: which treatment to offer?]. PMID- 10370551 TI - [Various modes of viral transmission from the mother to the fetus]. PMID- 10370552 TI - [Means of diagnosing viral infections in the fetus]. PMID- 10370554 TI - [Current status of clinical knowledge, physiopathology, and treatment of Langerhans histiocytosis (histiocytosis X)]. PMID- 10370553 TI - [Maternal-fetal viral infections: preventive treatment of maternal-fetal transmission during pregnancy, delivery and after birth]. PMID- 10370555 TI - [Genetic defects in control of T cell activation evolving in a syndrome of lymphohistiocytic activation]. PMID- 10370556 TI - [Hereditary abnormalities of phagocytic cells]. PMID- 10370557 TI - [Substitution therapy for deficiencies in antibody production by immunoglobulins]. PMID- 10370558 TI - [Mechanisms for treatment: periodic disease]. PMID- 10370559 TI - [Is it wrong to use immunostimulants in pediatric practice?]. PMID- 10370560 TI - [Community pneumopathies]. PMID- 10370561 TI - [Pneumopathies in children in developing countries]. PMID- 10370562 TI - [Nosocomial bacterial pneumopathies in children]. PMID- 10370563 TI - [Pulmonary infections in the newborn: diagnostic and therapeutic aspects]. PMID- 10370564 TI - [The infant, his mother and the doctor: the medicalization of infancy (18th-20th Century)]. PMID- 10370565 TI - [Emergencies as revealing asymmetries and paradoxes in the mother-caregiver relationship]. PMID- 10370566 TI - [Taking into account the health needs of the child in an emergency situation: the beginnings of the local approach]. PMID- 10370567 TI - [The increase in pediatric emergencies]. PMID- 10370568 TI - [The evolution of pediatric emergencies at Sainte-Justine Hospital]. PMID- 10370569 TI - [Emergency in pediatric hematologic oncology. French Group for the Study of Cancers and Leukemias of the Child (CFECLE)]. PMID- 10370570 TI - [Methods of recourse in pediatric emergencies: how is the activity inscribed in the field of ambulatory care?]. PMID- 10370571 TI - [The false debate about false emergencies]. PMID- 10370572 TI - [Participation of liberal pediatrics in the management of emergencies in France]. PMID- 10370573 TI - [Demography of pediatric medicine and emergencies]. PMID- 10370574 TI - [Child psychiatric emergencies: experiences in the creation of a reception and an emergency unit within CHU in Bicetre]. PMID- 10370575 TI - [Collaboration of child psychiatric nurses with pediatric emergencies]. PMID- 10370576 TI - [Analysis of methods of recourse for care and of the quality of management of children consulting for asthma or pediatric emergencies]. PMID- 10370577 TI - [Pediatric emergencies in the regional health program (SROS) Aquitaine]. PMID- 10370578 TI - [Project of a care network for the reception of medicosurgical pediatric emergencies in the urban community of Lyon]. PMID- 10370579 TI - [The place of interns: before, now and after...]. PMID- 10370580 TI - [Irradiation in the diagnostic design in pediatrics]. PMID- 10370581 TI - [Blood products and substitutes]. PMID- 10370582 TI - [Pediatric cardiology at the turn of the millennium: how can one control a cultural revolution?]. PMID- 10370583 TI - [The pathology of receptors: a concept which revives endocrinology]. PMID- 10370584 TI - [Artificial respiration in the child: the gains of the last five years]. PMID- 10370585 TI - [Friedreich's ataxia and mitochondria: the puzzle reconstructed]. PMID- 10370586 TI - [Diagnosis of states of ketosis in pediatrics]. PMID- 10370587 TI - Concomitant splenectomy for hypersplenic thrombocytopenia in hepatic resection for hepatocellular carcinoma. AB - BACKGROUND/AIMS: Resection of hepatocellular carcinoma (HCC) in patients with liver cirrhosis and hypersplenic thrombocytopenia (HSTC) is risky. Controversy exists concerning the role of concomitant splenectomy for HSTC in cirrhotic patients undergoing hepatectomy for HCC. METHODOLOGY: During the past 10 years, 294 patients have undergone hepatic resection for HCC in our department. Among them, 11 cirrhotic patients with severe HSTC (platelet count < or = 80000/mm3) underwent splenectomy simultaneously. The clinical outcomes were retrospectively reviewed. RESULTS: The resected spleen weighed 479 +/- 242 g. The post-operative mortality and morbidity were 9.1% and 27.3%, respectively. In all patients, the platelet count was elevated to above 100000/mm3, and serum total bilirubin was significantly lowered within 1 week of operation. The overall 5-year actuarial and disease-free survival rates were 66.7%. None of the patients developed severe infectious complications during the follow-up period. CONCLUSIONS: Concomitant splenectomy for severe HSTC in cirrhotic patients undergoing hepatectomy for HCC is justified as the benefits of concomitant splenectomy by far surpass the adverse effects. PMID- 10370588 TI - Is hepatic resection absolutely contraindicated for hepatocellular carcinoma in Child-Pugh class C cirrhotic patients? AB - BACKGROUND/AIMS: Liver resection for hepatocellular carcinoma (HCC) in Child-Pugh class C cirrhotic patients is considered to be high risk and even contraindicated. This study examined our results of hepatectomy for HCC in such cirrhotic patients. METHODOLOGY: A retrospective review of the clinicopathological features, as well as early and late resection results of Child-Pugh class A (n = 181) and class C patients (n = 13) were compared. The extent of hepatectomy was based on the pre-operative liver function test and indocyanine-green (ICG) clearance rate. RESULTS: The tumor size in class C patients was smaller than that in class A patients. There were no significant differences with regard to operative blood loss, amount of blood transfusion, operative morbidity or mortality. The surgical margins of class C patients were narrower (p = 0.003). The tumors of class C patients had higher incidences of well-formed capsules and absence of satellite nodules. The 5-year disease-free and actuarial survival rates of class A and C patients were 35.4% and 40.7% (p = 0.28), and 48% and 50% (p = 0.13), respectively. CONCLUSIONS: Not all HCCs in Child-Pugh class C cirrhotic patients are contraindicated for liver resection. In the absence of uncontrollable ascites, marked jaundice and encephalopathy, surgical resection is still justified in some selected cases, in spite of a narrow surgical margin. PMID- 10370589 TI - Surgical resection of hepatocellular carcinoma in elderly cirrhotic patients. AB - BACKGROUND/AIMS: Both cirrhosis and old age have been reported to be risk factors for hepatic resection. This study evaluated the clinical results of hepatic resection in elderly hepatocellular carcinoma (HCC) patients with cirrhosis. METHODOLOGY: During a 5-year period, 248 patients with HCC underwent curative hepatic resection. Among them, 24 elderly patients (age: > or = 70 years) with cirrhosis (Group I), 24 patients (age: > or = 70 years) without cirrhosis (Group II), and 98 patients (age: < 70 years) with cirrhosis (Group III) were selected for the study. The clinical and pathologic parameters, including pre-operative demographic features, surgical factors, pathological factors, DNA flow-cytometric analysis of the resected specimen, and post-resection prognosis were compared among the three groups. RESULTS: Group I patients had a significantly higher incidence of small-size tumors, hepatitis C infection, concomitant esophageal varices, and minor resection with a shorter surgical margin in the resected specimen. The surgical morbidity and mortality of Group I was similar to that of Group II and III patients. However, the disease-free survival rate was significantly lower in the Group I patients than in Group II (p = 0.02) and Group III patients (p = 0.04). CONCLUSIONS: Our findings indicate that although hepatic resection can be done safely in elderly cirrhotic HCC patients, the prognosis for these patients was less favorable even when curative resection was performed. PMID- 10370590 TI - Role of angiogenesis in hepatitis and hepatocellular carcinoma. AB - BACKGROUND/AIMS: Hepatocellular carcinoma (HCC) is usually a hypervascular tumor. Factor VIII-related antigens, including von Willebrand factor, are known to be expressed in HCC, which cause capillarization of the sinusoids of HCC. Capillarization of hepatic sinusoids may play a role in hepatocarcinogenesis and its metastasis. The aim of this study is to clarify the expression of Factor VIII in patients with hepatitis B or C (n = 18) and HCC (n = 16). METHODOLOGY: All specimens were sufficient for immunohistochemical study of the neo-angiogenesis with regard to clinical results. Microvessel count per square millimeter (MVC) and hot spot of microvessel per square millimeter (HSV) were measured from the histochemical study. RESULTS: In the patients with hepatitis group, the positive staining on the vessels of the portal triad was 11.1% (2/18) but in the non neoplastic tissue of HCC patients the positive rate was 68.7% (11/16) showing a significant difference from the hepatitis group. The amount of vasculatures was easily found in the surrounding capsule of resected HCC. The MVC of the capsule was 10.17 +/- 2.78 and 13.66 +/- 5.42 for the HCC with non-direct invasion and direct invasion during operation, respectively. The HSV of capsules were 7.51 +/- 2.09 and 9.14 +/- 4.02 for the non-invasion and invasion, respectively. Therefore, in our study, it is clear that the high MVC or HSV scores were found in patients of direct invasion. However, there was no relation between hepatitis B or hepatitis C to the tumor invasiveness. The median survival times were 21.5 months for the non-invasive group and 14.5 months for the invasive group (p < 0.05). The positive rate of Factor VIII in the vessels of the portal triad were 60% and 83.3% for the non-invasive and invasive groups, respectively. However, the lower values of MVC and HSV showed a trend toward a longer recurrence time. CONCLUSIONS: It is pertinent to prove that the high score of neo-angiogenesis has a high risk of recurrence. In addition, it is wise to pay more attention to the interval of the follow-up study to detect the recurrent lesion earlier, where possible, in the patient with a high score of microvascularity. PMID- 10370591 TI - Comparison of liver resection for hepatocellular carcinoma in hepatitis B and hepatitis C-related cirrhotic patients. AB - BACKGROUND/AIMS: The differences of liver resection for hepatocellular carcinoma (HCC) between hepatitis B and C-related cirrhotic liver remain unknown. This study compares the surgical results of HCC in hepatitis B and hepatitis C-related cirrhotic patients in an area endemic of hepatitis B. METHODOLOGY: A retrospective comparison of the clinicopathological features and early and long term results of 110 cirrhotic patients with seropositive hepatitis B surface antigen only (group B) and 55 patients with seropositive anti-hepatitis C antibody only (group C) was carried out. RESULTS: Group C patients were older, had a lower serum alpha-fetoprotein level, greater indocyanine retention rate, and higher incidence of multicentric tumors. Tumor size was larger and there was a higher incidence of combined satellite nodules in group B patients. There were no significant differences in operative morbidity and mortality between the two groups. Group B patients had a slightly shorter disease-free interval (p = 0.07) but a better actuarial survival rate (p = 0.05) than group C patients. CONCLUSIONS: The hepatitis status did not affect the operative risks in cirrhotic livers. However, after resection of HCC, poorer liver functional reserve in hepatitis C-related cirrhotic patients caused poorer actuarial survival rate when compared with hepatitis B-related cirrhotic patients. PMID- 10370592 TI - Dysplasia in the gastrointestinal tract. AB - Epithelial dysplasia is an unequivocal neoplastic transformation which can be recognized with routine morphology. It is the morphological expression of one of the early steps of carcinogenesis. Its diagnosis depends upon the recognition of cytological and architectural abnormalities. A dysplastic lesion can, according to the severity of these abnormalities, be subdivided into different degrees. The presence of dysplasia in endoscopic biopsies may indicate a precursor lesion but it may also be a marker of malignancy. New techniques looking for molecular alterations can help to make a more reliable diagnosis of dysplasia. PMID- 10370593 TI - Molecular and phenotypic markers of hamartomatous polyposis syndromes in the gastrointestinal tract. AB - Hamartomatous gastrointestinal polyposis syndromes have always been considered as non-neoplastic. Nevertheless, an increased cancer risk both within and outside the gastrointestinal tract may exist in these syndromes. The hamartomatous polyps may sometimes harbor dysplasia, but their neoplastic potential is unknown. The genetic defects causing the hamartomatous syndromes are less well established than, for example, familial adenomatous polyposis (FAP) and hereditary non polyposis colorectal cancer (HNPCC). The genetic studies on the Mendelian inherited syndromes FAP and HNPCC have made a major contribution to the identification of genes involved in colorectal tumorigenesis. The genes involved in colorectal cancer development may also contribute to cancer development in the hamartomatous polyposis syndromes, and are currently under investigation. Furthermore, new insights into the development of various cancers may be obtained by the isolation and characterization of genes involved in Mendelian inherited hamartomatous polyposis syndromes. This report summarizes the available literature on this subject, and describes the pheno- and genotypic features of the hamartomatous syndromes of juvenile polyposis, Peutz-Jeghers syndrome, and Cowden's disease. PMID- 10370594 TI - Upper gastrointestinal polyps in familial adenomatous polyposis. AB - Familial adenomatous polyposis (FAP) is an autosomal dominant disease in which affected family members develop numerous colorectal adenomas with a virtually 100% chance of malignant degeneration unless the colon is prophylactically removed. After colectomy the main cause of death is upper gastrointestinal malignancy. The majority of FAP patients also develop upper gastrointestinal polyps, and especially those in the antrum and duodenum are usually neoplastic. Therefore, surveillance of FAP patients through endoscopy plus biopsy is recommended. The Spigelman classification in which the number of adenomas, the size, architecture and degree of dysplasia account for the scoring system, provides a guide for follow-up in these patients. Molecular genetic markers to assess the risk of upper gastrointestinal cancer in FAP patients are as yet not available. PMID- 10370595 TI - Cell kinetic measurements: principles, guidelines for treatment? PMID- 10370596 TI - Pancreatic endocrine tumors: diagnostic pitfalls. PMID- 10370597 TI - Radioimmunoguided surgery for colorectal carcinoma. PMID- 10370598 TI - Mechanisms and prevention of recurrent colorectal cancer. PMID- 10370599 TI - The medical treatment of colorectal cancer: actual status and new developments. AB - Colorectal cancer is one of the most frequent malignancies and one of the greatest causes of cancer death in the Western world. The prognosis is determined by the stage at diagnosis. Patients with metastatic colon cancer have a bad prognosis. Chemotherapeutic treatment with 5-Fluorouracil (5-FU) and folinic acid is actually considered as the standard treatment in patients with metastatic disease. Although the survival benefit is relatively small, many patients can benefit from this treatment in terms of tumor regression or symptom improvement. Several new drugs are actually in development and create hope for improved tumor or symptom control and longer survival. Thymidylate synthase inhibitors (raltitrexed), topoisomerase I inhibitors (irinotecan), the oral 5-FU prodrugs (capecitabine, UFT), ethynyluracil, and oxaliplatin are promising new drugs. The challenge will be to determine the best combination of these new drugs and the exact sequence in which these drugs will be used. Adjuvant post-operative chemotherapy in colon cancer is one of the most important advances in oncology that has been introduced into the clinic during the last years. For rectal cancer, an adjuvant treatment should consist of a combined chemo-radiotherapy. The search for better prognostic factors for recurrence should help to focus on a better adjuvant treatment for patients with the highest risk for recurrence. PMID- 10370600 TI - Reflections on three field lymphadenectomy in carcinoma of the esophagus and gastroesophageal junction. AB - BACKGROUND/AIMS: One of the most controversial questions in the surgical treatment of carcinoma of the esophagus and gastroesophageal junction (GEJ) is the extent of lymph node dissection, in particular the value of the cervical lymph node dissection (the so-called third field). METHODOLOGY: This study reflects a single institution's experience with this extensive lymphadenectomy, the technique of which is described in detail. RESULTS: Adding the third field to the lymph node dissection markedly improved accuracy of staging. Unforeseen involvement of lymph nodes in the neck was found in 30%. In T3N+ tumors of the GEJ, as much as 16.6% of positive lymph nodes were detected in the neck. Locoregional recurrence without distant metastasis was found in 6 patients (17.8%) out of a group of 37 patients with a minimum follow-up of 5 years. All 6 patients had stage IV disease because of distant lymph node metastasis (M+Ly). In 3 of these patients, locoregional recurrence occurred only after 3 years or more. In a subsequent series of 100 esophagectomies performed between 1992 and July 1993 no difference in outcome between radical versus standard resection was noticed for early stage I and II. However, there is a tendency towards a better estimated 5-year survival in favor of radical dissections (21%) versus standard resection (12%) in stage III and IV. CONCLUSIONS: Extensive three field lymphadenectomy can be safely performed without increasing hospital mortality (0%) and morbidity. Improved accuracy of staging, prolonged disease-free survival and potential increased cure rate are confirmed by our experience. Survival obtained with this technique has to be compared with survival obtained by other, multimodality treatment forms. PMID- 10370601 TI - Risk factors of acute ulcer bleeding. AB - Peptic ulcer disease is the most common cause of upper gastrointestinal bleeding. Most ulcers stop bleeding spontaneously; however, a poorer prognosis is indicated by clinical features such as severe bleeding, inability to clear gastric lavage, advanced age, and serious co-morbid illness. NSAID usage is an important risk factor particularly in the elderly population. Endoscopic stigmata of recent bleeding may provide the most helpful prognostic information which helps identify high risk patients and direct management specifically at preventing rebleeding in this cohort of patients. PMID- 10370603 TI - Endoscopic therapy for upper gastrointestinal hemorrhage: a state of the art. PMID- 10370602 TI - The role of endoscopic Doppler-sonography. AB - Endoscopic Doppler sonography is a useful method that permits a differentiation to be made between high-risk lesions in danger of rebleeding and prognostically harmless ulcerations. Using this technique, vessels in the base of the ulcer can be reliably identified, and the indication for local endoscopic treatment established. The pulsed Doppler can be used to test the efficacy of prior endoscopic therapy within the framework of follow-up investigations; when arterial blood flow signals are found to persist, treatment needs to be repeated. With the aid of this "programmed" Doppler sonography-controlled endoscopic approach, which in some cases may be repeatable, the number of rebleeds, emergency operations, and probably also mortality, can be permanently lowered. Endoscopic Doppler sonography can also provide important additional information in the area of primary diagnosis and endoscopic treatment of esophageal and gastric varices. The Doppler exploration facilitates the assessment of the sclerosing effect, and is capable of identifying gastric varices and distinguishing these from other submucosal processes. For an assessment of the butyl cyanoacrylate varix, the Doppler is of particular value. PMID- 10370604 TI - Non-surgical treatment of variceal bleeding. AB - Variceal bleeding is still one of the key therapeutic problems inpatients with portal hypertension. Vasoactive drugs for the treatment of acute hemmorrhage as well as for the prevention of first and recurrent bleeding have been shown to be effective, in some situations as effective as endoscopic sclerotherapy. In recent years, endoscopic band ligation has replaced sclerotherapy for the prevention of rebleeding and become the new nonsurgical standard in this situation. Furthermore, the transjugular, intrahepatic, portosystemic stent shunt (TIPS) has gained widespread acceptance as a salvage procedure for acute bleeders unresponsive to other treatments as well as the second line approach to failures of chronic endoscopic or drug therapy for prevention of rebleeding. Primary prophylaxis is still the domain of beta-blockers or/and long-acting nitrates. PMID- 10370605 TI - Laparoscopic versus open treatment of patients with acute cholecystitis. AB - BACKGROUND/AIMS: The studies published so far mention a high rate of complication and conversion in laparoscopic surgical treatment of acute cholecystitis. Considering the relatively high conversion rate in cases of acute cholecystitis, it is necessary to pre-operatively estimate the chance of successful laparoscopic cholecystectomy. One of the aims of this study was to determine the factors that influence the chance of success of this technique. Another aim was to define possible advantages of the method. METHODOLOGY: From 1991 through to 1995, a total of 295 patients in whom acute cholecystitis had been diagnosed on the basis of clinical examination, laboratory data, ultrasonography and pathohistological examination, underwent operative therapy. The laparoscopic approach was attempted in 49 of these patients. Since the patients who underwent primary open surgery were markedly handicapped with regard to severity of inflammation and co-morbid factors, we identified a sub-group of these patients who were comparable to those who underwent laparoscopic cholecystectomy in accordance of the above-mentioned criteria. RESULTS: The rate of conversion (44.9%) correlated with the severity of inflammation, which was determined on the basis of leukocytosis > 10 x 10(9)/l (p = 0.004) and the pathohistological diagnosis (p = 0.005). Hence, the rate of conversion was 71.4% in cases of empyema of the gallbladder but only 29.2% in cases of edematous cholecystitis. In patients whose leukocyte count decreased within 4 days of conservative treatment, a successful laparoscopic cholecystectomy (LC) was performed in 91.7% (11/12) of cases, while 8 patients whose leukocyte count increased or showed no reduction during this time required conversion to open cholecystectomy (p = 0.0001). In cases of acute cholecystitis, the complication rate after LC is lesser in respect of wound infection (p = 0.07) and pneumonia (p = 0.04). In all patients, obesity was a risk factor for wound infection (p = 0.04). Injury to the small intestine was registered in 1 case but in no case was LC associated with injury to the bile duct. CONCLUSIONS: The degree of inflammation and its response to conservative treatment, which are determined on the basis of leukocytosis and clinical improvement, are clear indications of the chance of successful delayed laparoscopic cholecystectomy within the first week. Hence, all patients whose leukocyte count does not decrease after antibiotic treatment should be treated with open cholecystectomy (OC). The complication rate following LC is less than that following OC. Although no injury to the bile duct has been observed in cases of acute cholecystitis, major complications are possible and should not be excluded. PMID- 10370606 TI - Enhanced dissolution of gallstone by combining ethanol with two commonly used cholelitholytic solvents. AB - BACKGROUND/AIMS: Contact dissolution therapy is one of the non-surgical treatments for patients with gallstone. Among the various solvents, methyl tert butyl ether (MTBE) is used for cholesterol gallstone, while tetrasodium ethyl dimethyl tetraacetate (EDTA-4Na) solution is used to dissolve calcium bilirubinate stones. However, the contents of gallstone cannot be precisely predicted while they are still present in the human body. This study was designed to test if the MTBE and EDTA can be mixed together and to test the solubility of different kinds of gallstone in each original solution and mixture. METHODOLOGY: Each 0.1 gm of mixed cholesterol stone, brown stone and pigment stone from 18 patients was used. Pure ethanol was chosen to enhance the miscibility between the organic phase of MTBE and the aqueous phase of EDTA. The contents of gallstone after dissolution were examined with scanning electromicroscopy. RESULTS: We found the mixture of ethanol, MTBE and EDTA to be the most efficient solvent in gallstone dissolution in comparison with the other two original solvents. The mixture reached a dissolution percentage of 97.96 +/- 1.00, 88.96 +/- 6.51 and 67.75 +/- 14.26 for cholesterol, brown and black pigment gallstone, respectively. CONCLUSIONS: We concluded that ethanol can be used to mix the MTBE and EDTA with good preservation in their litholytic effects on gallstone. The ethanol-MTBE-EDTA solvent is, therefore, a promising universal cholelitholytic agent which deserves further tests for its safety and efficacy in the in vivo study. PMID- 10370607 TI - Endoscopic nasobiliary drainage for treating bile leak after laparoscopic cholecystectomy. AB - BACKGROUND/AIMS: Bile leaks are common complications of laparoscopic cholecystectomy. We evaluated the diagnosis and endoscopic treatment of bile leaks. METHODOLOGY: A total of 436 patients underwent laparoscopic cholecystectomy with infrahepatic drainage. We performed immediate endoscopic retrograde cholangiopancreatography (ERCP) on all patients with bile discharge from an infrahepatic drain, and treated bile leaks which were not due to a major ductal injury by endoscopic nasobiliary drainage (ENBD) without endoscopic sphincterotomy (ES). RESULTS: Ten patients developed bile leaks which were recognized within 18 hours of operation. ERCP, on post-operative day 1 or 2, showed a bile leak from the cystic duct (9 patients) or the liver bed (1 patient). All patients underwent ENBD. Only 1 patient, who had a retained stone, had ES. In all patients, the bile leak resolved promptly and both the infrahepatic and nasobiliary drains were removed within 6 days of cholecystectomy. All patients were asymptomatic at a mean follow-up of 30 months. CONCLUSIONS: Routine placement of an infrahepatic drain is recommended for the early detection of bile leaks. Bile leaks can be successfully treated by prompt ENBD without ES. PMID- 10370608 TI - Serum alkaline phosphatase after extensive liver resection: a study in patients with biliary tract carcinoma. AB - BACKGROUND/AIMS: To clarify a correlation between serum alkaline phosphatase (ALP) levels and liver function and regeneration after major hepatectomy. METHODOLOGY: Post-operative changes in serum ALP levels were retrospectively examined in 91 non-cirrhotic patients with biliary tract carcinoma who underwent right hepatic lobectomy or more extensive liver resection. In addition, changes in liver volume after resection were assessed in 31 patients who underwent computed tomography before surgery and within 1 month after resection. RESULTS: Serum ALP levels reached its nadir on post-operative day 1, followed by a gradual increase until post-operative day 28. In patients with post-hepatectomy liver failure (n = 32), serum ALP levels were significantly lower on days 1, 7, 10, 14, 21, and 28 after resection than in those without such failure (n = 59). Unexpectedly, the volumetric study of the liver showed no significant difference between the two groups in the remnant liver volume after resection. CONCLUSIONS: Serum ALP levels can function as an indicator of liver function after hepatectomy, but not reflect morphological regeneration of the liver. Thus, increased ALP levels after hepatectomy may not reflect the cellular proliferation process itself. PMID- 10370609 TI - Percutaneous drainage of emphysematous cholecystitis associated with pneumoperitoneum. AB - Emphysematous cholecystitis, a relatively rare variant of acute cholecystitis, is associated with high morbidity and mortality rates. In the presence of a concomitant pneumoperitoneum, these rates may be considered even higher, approaching those of perforation of the gallbladder. The first choice of treatment in cases presenting with pneumoperitoneum is emergency laparotomy. We performed a staged procedure as a second best alternative. In a 65 year-old female patient, initial percutaneous cholecystostomy with a strict intravenous antibiotics regimen, and subsequent cholecystectomy 6 months, later was carried out with successful outcome. A review of the literature revealed 13 other cases of this combination. Treatment modalities and outcome of these patients are discussed. PMID- 10370610 TI - Hyperbaric oxygenation as adjuvant therapy to surgery of emphysematous cholecystitis. AB - Three cases of emphysematous cholecystitis are presented. The role of hyperbaric oxygenation as excellent adjuvant therapy to urgent surgical as well as intensive conservative treatment is emphasized. PMID- 10370611 TI - The role of lymphatic drainage of the liver in gallbladder cancer: a case report. AB - We report a case of a patient with a unique lymph node relapse after right hepatectomy and aggressive lymph node dissection for gallbladder cancer. There was extensive involvement of the hepatic parenchyma from the primary tumor, but no extension to the lymph nodes or other adjacent organs. Seventeen months later, the patient underwent re-dissection of the retroperitoneal lymph nodes with right nephrectomy and partial resection of the vena cava because of lymph node recurrence at the hilum of the right kidney. This pattern of lymph node metastasis to the right side of the vena cava from gallbladder cancer invading the liver is probably due to the distinct lymphatic drainage of the liver. PMID- 10370612 TI - Aneurysm of the celiac trunk: diagnosis with US-color-Doppler. Presentation of a new case and review of the literature. AB - Aneurysms of the celiac trunk are the rarest forms of aneurysms of the visceral arteries. Since 1958, when Schumaker reported the first case to be successfully treated surgically, only 69 cases have been reported in the international literature. The detection of such aneurysms, which are often asymptomatic, is mostly occasional. Approximately 15-20% of cases may be complicated by rupture with a mortality rate of around 80%. This eventuality makes surgical treatment mandatory even in asymptomatic cases. The authors report on their experience with the surgical treatment of one case of aneurysm of the celiac trunk and then go on to review the relevant literature. PMID- 10370613 TI - The impact of genetics on Crohn's disease. AB - BACKGROUND/AIMS: Reports focusing on the familiarity and "pedigree" of patients with Crohn's Disease (CD) are increasing. The study of the role of genetics as a predisposing factor in providing the ideal milieu upon which environmental agents and immuno-inflammatory responses may act, could be paramount in finding the pith of the etiopathogenesis of this disease. METHODOLOGY: In order to determine the impact of familiarity on CD, a series of 187 patients, managed between January 1965 and January 1997, was subdivided into two groups. In group I (145 pts.), the family history relied only on direct information from the patient, while in group II (42 pts.) a prospective study was carried out involving both close and distant relatives who were interviewed and, in some cases, clinically investigated. RESULTS: In this study, relatives with suspected CD were 9 out of 122 in group I patients (7.4%), while in the more detailed assessment of group II, 18 out of 42 cases (43%) had an ascertained CD familiarity. CONCLUSIONS: The importance of familiarity in the pathogenesis of CD may be higher than expected if properly sought for. Reports on the possibility that the onset of CD may be strongly influenced by genetics, favor our hypothesis that the true etiology may find its base in a hemolymphatic disorder of the mesentery followed by superimposed inflammatory responses. PMID- 10370614 TI - Endoscopic sphincterotomy in the management of biliary leakage after partial hepatectomy. AB - We present the case of a 22 year-old man with biliary leakage caused by partial hepatectomy. He was successfully treated with endoscopic sphincterotomy alone. PMID- 10370615 TI - Stop-flow thoracic perfusion for unresectable pulmonary carcinoma with or without inhalation IL-2 immunotherapy. A prospective randomized study. AB - BACKGROUND/AIMS: Pulmonary carcinoma, metastatic or primary, remains a challenging disease with a poor and dismal prognosis. This prospective randomized study compares the results of stop-flow thoracic perfusion and systemic chemotherapy with or without the addition of Interleukin-2 (IL-2) inhalation immunotherapy with respect to overall survival and quality of life. METHODOLOGY: From January 1996 to January 1999 stop-flow thoracic perfusion using two balloon tipped catheters was carried out in combination with maintenance system chemotherapy, with (Group B) or without (Group A) inhalation IL-2 immunotherapy in 67 patients suffering from histologically diagnosed unresectable lung carcinoma. RESULTS: The mean survival for Group A patients was 12 months (M +/- SD: 12 +/- 8.82) while in Group B patients it was 19 months (M +/- SD: 19 +/- 8.47). CONCLUSIONS: Stop-flow thoracic perfusion in combination with systemic maintenance chemotherapy and IL-2 inhalation immunotherapy prolongs survival and ensures a good quality of life. PMID- 10370616 TI - Transplenic and transtumoral in vivo immunostimulation: effect on cellular immunity parameters. AB - BACKGROUND/AIMS: It has been shown that IL-2 and gamma-IFN own an additive immunostimulatory effect against tumors. Moreover, it has been suggested that the locoregional route of administration of immunopotentiating agents has substantial advantages compared to the systemic route. The aim of the present study was to evaluate the immune response of patients with advanced unresectable liver and pancreatic malignant disease who were submitted for transplenic and transtumoral IL-2 and gamma-IFN combination immunotherapy. METHODOLOGY: In 32 patients, the following therapeutic regimen was applied for 10 consecutive days: a) 5 days transplenic administration of 5 mg IL-2 plus 50 micrograms gamma-IFN in a Lipiodol-Urografin emulsion, and b) 5 days transtumoral administration of the same drug cocktail. In the beginning (T1) and on day 15 (T2) after treatment, the following parameters were performed: CD3+, CD4+, CD8+, IL-2r+, HLA-DR+ lymphocytes in peripheral blood spontaneously, and after 48-hour T-lymphocyte cultures in the presence of PHA. RESULTS: CD3+, CD4+ and CD8+ T-lymphocytes were significantly increased in peripheral blood after treatment (p < 0.05, p < 0.05, and p < 0.003, respectively), whereas IL-2r expression and HLA-DR expression on CD4+ T-cells were not influenced significantly (p > 0.05). After PHA 48-hour T cell cultures, T-cell subpopulations showed the following alterations: CD3+, CD4+ and CD8+ T-cells were significantly increased (p < 0.001, p < 0.006 and p < 0.01, respectively). Moreover, IL-2r expression as well as HLA-DR expression on CD4+ T cells were also significantly increased (p < 0.001). CONCLUSIONS: Locoregional treatment exerted a beneficial immunopotentiating effect on T-cell populations, IL-2r expression and HLA-DR expression. This is a promising mode of action in the management of malignant metastatic disease. PMID- 10370617 TI - Detection of bile duct cancer by autofluorescence cholangioscopy: a pilot study. AB - BACKGROUND/AIMS: There is increasing interest in the detection of malignancy by autofluorescence endoscopy. This is the first report on autofluorescence endoscopy in bile duct cancer. METHODOLOGY: Nine patients with bile duct cancer underwent percutaneous transhepatic autofluorescence cholangioscopy using light induced fluorescence endoscopy in the gastrointestinal tract (LIFE-GI) system. RESULTS: The cancerous lesion had quite a different color in comparison with normal mucosa, which was seen as light blue, by the autofluorescence endoscopy. In all patients, cancerous legions were observed as dark red. Additionally, in 7 of the 9 cases a white fluorescence was also seen in the cancerous lesion. CONCLUSIONS: In previous reports on the gastrointestinal tract and bronchial tree, cancerous lesions appeared dark red when examined by autofluorescence endoscopy. In our study, it is suggested that a white fluorescence is also an important finding for making diagnosis of bile duct cancer by the autofluorescence endoscopy. PMID- 10370618 TI - Retained foreign bodies following intra-abdominal surgery. AB - BACKGROUND/AIMS: A retained foreign body in the abdominal cavity following surgery is a continuing problem. Despite precautions, the incidence is grossly underestimated. The purpose of this study is to report the result of surgical treatment on 24 consecutive cases treated by the authors during a 10-year period. METHODOLOGY: All consecutive patients with a confirmed diagnosis of foreign body after abdominal surgery were studied. Data collected included the patients' age and sex, the initial diagnosis and primary surgical treatment, period of time between the probable causative operation and the definitive treatment, nature of the foreign body, clinical presentation, predisposing factors, and diagnosis and management; morbidity and mortality are presented as well as guidelines for prevention. RESULTS: All patients were symptomatic. Eight patients presented as intraabdominal sepsis (4 with intestinal obstruction, 4 with entero- or colo cutaneous fistula), non-specified abdominal pain in 3, persistent sinus and granuloma in 2, abdominal palpable mass in another 2 cases, and 1 patient with vaginal discharge. The diagnosis was established pre-operatively in 15 cases by means of plain abdominal radiographs, ultrasound or computed tomography (CT) scan. Morbidity was observed in 50% and the rate of surgical reinterventions because of fistulas or residual sepsis in 18%. The mortality was almost 10%. CONCLUSIONS: The clinical manifestations ranged from mild abdominal pain, palpable mass, persistent drainage and granuloma to intestinal obstruction secondary to adhesions or occlusion of the intestinal lumen because of migration of the foreign body and intraabdominal sepsis. Despite this being a rare situation, when it happens it presents as a very serious problem to patients with high rates of morbidity and mortality. Prevention remains the key to the problem. PMID- 10370619 TI - Cell proliferation and cell loss in nodule-in-nodule hepatocellular carcinoma. AB - BACKGROUND/AIMS: In order to clarify the significance of the balance between cell proliferation and cell loss during the progression of hepatocellular carcinoma, 16 operative specimens of nodule-in-nodule hepatocellular carcinoma were investigated. METHODOLOGY: In 16 specimens, cell proliferation was evaluated by the expression of Ki-67 nuclear antigen, and cell loss was also examined by the method of terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL). The expressions of p53 protein, bcl-2 protein and Fas antigen were also investigated to clarify the relationship between their expression and cell kinetics. RESULTS: The Ki-67 labeling index of the inner nodules was higher than that for the outer nodules (18.9% vs. 7.2%; p < 0.05) and the TUNEL labeling index of the inner nodules was also higher than that for the outer nodules (12.8% vs. 6.6%; p < 0.05). The increasing rate of the Ki-67 labeling index from Edmondson's grade I to II was 3.9 +/- 3.0, that from grade II to III was 3.9 +/- 2.4, while the increasing rate of the TUNEL labeling index from grade I to II was 2.7 +/- 0.3 and that from grade II to III was 1.7 +/- 0.2 (p < 0.05). p53 Protein was observed in 5 cases, while bcl-2 protein was found in 4 cases in the border area of the inner nodule. However, Fas antigen was found in none of the examined cases. Regarding the Ki-67 positive rate in the inner nodule, the Ki-67 positive rate in the p53 protein positive cases was significantly higher than that in the negative cases (30.3 +/- 15.4 vs. 11.9 +/- 9.2; p < 0.05). However, the TUNEL labeling index was not affected by the expression of those proteins. CONCLUSIONS: This study suggested that tumor progression depends on a disturbance in the cell kinetic balance caused not by a decrease in the absolute amount of cell loss but in the chaotic balance between cell loss and cell proliferation. PMID- 10370620 TI - Laparoscopic splenectomy: the latest modern technique. AB - BACKGROUND/AIMS: Recent advances in technical instruments have resulted in increased safety and simplicity in laparoscopic surgery. The purpose of this article is to introduce our latest operative techniques for laparoscopic splenectomy. METHODOLOGY: The patient is placed in the right semidecubitus position and the gastrosplenic ligament including the short gastric vessels was performed by using an ultrasonically activated scalpel. The splenic artery and vein were resected at the splenic hilum with an autosuture device. The electromechanical morcellator was used to remove the spleen. RESULTS: The laparoscopic splenectomy was successfully performed in all 74 patients from 1992 1997. There was no deaths related to the operation. Conversion to open surgery with a small incision of 5 cm was required in one patient with advanced liver cirrhosis and portal hypertension and 45 patients with portal hypertension. CONCLUSIONS: A laparoscopic splenectomy is considered to be a safe and feasable modality for the treatment for hematologic disorders of both the spleen and other benign tumors. PMID- 10370621 TI - Gene transfer with cationic lipid into human hepatocellular carcinoma in nude mice. AB - BACKGROUND/AIMS: Using a cationic lipid, gene transfection into the tumor of a human hepatocellular carcinoma model in nude mice was attempted in order to explore the possibility of its use in gene therapy. METHODOLOGY: A DNA-lipid complex was made by combining the cationic lipid distearyldimethyl ammonium bromide (DDAB) with pCMV sPORT expressing the reporter gene LacZ. The expression of this complex was first investigated in vitro against the human hepatocellular carcinoma cell line Li7HM. It was then injected directly into a hepatocellular carcinoma model tumor created by implanting Li7HM into the liver of BALB/c nu/nu mice, and the expression of LacZ was histologically evaluated. RESULTS: LacZ gene was expressed in Li7HM in vitro with the optimized DNA-lipid complex. Cell toxicity was not a problem. Expression of LacZ was also seen in the mouse hepatocellular model tumor into which the complex had been injected, indicating successful gene transfection with this method. CONCLUSIONS: Direct injection of a DNA-lipid complex into a mouse hepatocellular carcinoma model tumor is a safe and simple method of gene transfection, proving this to be a viable method of transfer for use in gene therapy. PMID- 10370622 TI - Reappraisal of K-ras and p53 gene mutations in the recurrence of Dukes' B2 rectal cancer after curative resection. AB - BACKGROUND/AIMS: Recurrence of rectal cancer remains a major clinical problem. This study was conducted to evaluate the impact of K-ras and p53 mutations on the recurrence of rectal cancer. METHODOLOGY: A total of 166 resected Dukes' B2 stage rectal carcinomas were collected between January 1990 and April 1994. The stored frozen tissues were retrieved for immunocytochemistry of p53 and genomic analysis of K-ras and p53 genes. The data of K-ras and p53 gene mutations were correlated with clinicopathological variables. The concordance of immunocytochemistry with genomic analysis in the survey of p53-mutations was examined. The follow-up data were analyzed by Kaplan-Meier estimator. RESULTS: Sixty-nine patients (41.6%) developed recurrent tumor. A significantly higher recurrence rate (p = 0.0013) and shorter median recurrence time were noted in p53 mutated than non-mutated cancers. Mutations in K-ras gene do not significantly increase the risk of tumor recurrence (p = 0.1702). K-ras and p53 mutations are not associated with clinicopathological parameters (p > 0.05). Kappa statistic indicates highly significant reproducibility between immunocytochemistry and genomic analysis for p53 mutations (p < 0.0001). CONCLUSIONS: Presence of p53 mutation significantly increases the recurrence rate and shortens the recurrence time of the resected rectal cancers. Pre-operative routine check for p53 mutations by immunocytochemistry may be beneficial in choosing the optimal surgical strategy for rectal cancer. PMID- 10370624 TI - Surgery of icteric-type hepatoma after biliary drainage and transcatheter arterial embolization. AB - BACKGROUND/AIMS: The authors aimed to study the importance of pre-operative jaundice reduction in the surgical treatment of icteric-type hepatoma (IHCC). METHODOLOGY: A series of 10 patients with IHCC was reviewed. Eight out of the 10 patients underwent biliary drainage. Obstructive jaundice in the other 2 patients resolved spontaneously. Nine patients subsequently underwent transcatheter arterial embolization (TAE), which appeared to have an additional effect in reducing jaundice. RESULTS: Consequently, 9 of the 10 patients achieved sufficient reduction of the jaundice preoperatively. After the evaluation of liver function, 8 patients underwent hepatectomy without any appreciable morbidity or mortality. The median survival time of the resected cases was 18 months. CONCLUSIONS: A combination of biliary drainage and subsequent TAE is a recommended pre-operative strategy for the successful surgical treatment of IHCC. PMID- 10370623 TI - Liver, spleen and tumor volume measured by personal computer. AB - BACKGROUND/AIMS: Computed tomography (CT) scans are common examinations for patients with chronic liver diseases. To quantitate the organ or tumor volume from the scans and to accomplish the task in an efficient way with the most economic equipment, we developed a system based on a personal computer. METHODOLOGY: We used color-markers and transparency to sketch the edges of liver, hepatoma, and spleen. Each organ or tumor of interest is marked out by fine-point markers on pieces of transparency. The sketch was scanned into a digitized image format on a personal computer (Pentium 133). The calculation involves edge detection, three-dimensional reconstruction, and voxel counting. By using summation-of-the-area and trapezoid approximation technique, the voxels of each structure are counted. In this study, we illustrate the potential application in the management of a hepatic cancer patient. RESULTS: After digitalization, the data size of CT images is about 1 to 1.5 megabytes. It takes less than 5 min to complete volume calculation. CONCLUSIONS: By this method, tumor load before and after chemotherapy can be estimated easily and accurately. This would be helpful in clinical practice. PMID- 10370625 TI - Fundal varices: problem and management. AB - BACKGROUND/AIMS: The pathophysiology of gastric varices may be due to generalized or segmental portal hypertension. A considerable debate has arisen regarding the role of injection sclerotherapy in the pathogenesis of gastric varices. METHODOLOGY: During the period from 1987 to 1997, a total of 1686 cases with bleeding varices were presented to our center and 225 cases (13.3%) with bleeding gastric varices were diagnosed. There were 198 males and 27 females with a total mean age of 45.7 years (+/- 7.6). Primary fundal varices (FV) were found in 121 (54%) cases and secondary FV were found in 104 (46%) cases. All patients with isolated FV presented with repeated attacks of upper gastrointestinal bleeding. RESULTS: The pathological diagnosis was studied in 120 cases; it was schistosomal in 8.3% of cases, non-schistosomal in 33.3% of cases, and mixed (Schistosomal with post viral cirrhosis) in 58.3% of cases. Seventy-five cases were subjected to splenectomy and gastroesophageal decongestion (SGED), 64 cases were subjected to distal splenorenal shunt (DSRS), and 86 cases were subjected to sclerotherapy. Mortality after DSRS was 7.8%, after SGED it was 12%, and after sclerotherapy it was 21%. Rebleeding was the major complication and occurred in 3% after DSRS, in 13% after SGED, and in 18% of cases after sclerotherapy. CONCLUSIONS: Gastric varices are not an uncommon condition as a cause of upper gastrointestinal bleeding. Our findings support the hypothesis that gastric varices may be considered a late sequel of injection sclerotherapy, though they may also be considered as one of the pathophysiologies of generalized portal hypertension. Finally, DSRS was found to be the treatment of choice in the management of fundal varices. PMID- 10370626 TI - Glutaraldehyde-fixed heterologous pericardium for vena cava grafting following hepatectomy. AB - BACKGROUND/AIMS: Glutaraldehyde-fixed heterologous pericardium has been widely used for grafts in cardiac surgery. We applied it for inferior vena cava (IVC) patch grafting following combined resection of the liver and the IVC. METHODOLOGY: IVC grafting using a glutaraldehyde-fixed horse pericardium following combined resection of the liver and the IVC was performed in 2 patients -one with hepatocellular carcinoma and the other with hepatic metastasis following rectal cancer. The retrohepatic vena cava defect was closed with a 10 x 5 cm patch in one patient and a 7 x 4 cm patch in the other. RESULTS: Hepatic vascular exclusion was avoided in both patients. The IVC exclusion period was 40 min for the first patient and 25 min for the second. One patient required a veno venous bypass with an active centrifugal pump of 153 min. There was no complication and no graft infection. The microscopic extension to the IVC was evident in one patient, and fibrous adhesive was evident in the IVC wall of the other. One patient died of hepatic failure 3 years and 6 months after surgery, and the other died of hepatic recurrence 7 months after surgery. Both grafts were patent, without calcification and stricture, until the patients' death. CONCLUSIONS: Glutaraldehyde-fixed heterologous pericardium is an option for IVC grafting. PMID- 10370627 TI - Total gastrectomy. AB - Total gastrectomy with lymph node dissection is one of the standard operations for gastric malignancies without distant metastases. Surgical procedures of total gastrectomy are described with illustrations easy to understand anatomy. Practical application and survival of patients are also shown. PMID- 10370628 TI - US guided biliary drainage during hepatopancreatico-jejunostomy for diffuse bile duct carcinoma. AB - Liver failure is one of the principal causes of post-operative morbidity and mortality after major hepatectomy for diffuse bile duct cancer. To prevent this complication, biliary decompression must be guaranteed before and during the operation. If a nasobiliary catheter is positioned pre-operatively, biliary drainage can be maintained during hepatopancreato-duodenectomy by introducing a transhepatic drain under sonographic guidance. This original technique is described herein. PMID- 10370630 TI - Pancreatic head cystadenoma: a case report. AB - An 18 year-old resident of Zagreb was admitted to our hospital with intermittent pain in the right subcostal region. On examination, a palpable resistance was found in the upper abdomen. After extensive clinical and laboratory tests, a tumor of the pancreatic head, 80-85 mm in diameter, was verified. Cytologically, a diagnosis of microcystic adenoma of the pancreas was established. The patient underwent a cephalic pancreatoduodenectomy with preservation of the pylorus. Six months later the patient was no longer on a diet and, at follow-up, 3 years after surgery, she is symptom-free and feeling well. PMID- 10370629 TI - Recurrence of the Budd-Chiari syndrome after orthotopic liver transplantation. AB - Obstruction of the hepatic venous outflow with or without involvement of the vena cava results in the Budd-Chiari syndrome (BCS). BCS may be limited to the liver but there is a variety of systemic disorders forming the etiology of BCS in the majority of cases. Surgery has a major impact on treatment of the BCS within a wide range of therapeutic strategies. The ultimate option of surgical management of the BCS is orthotopic liver transplantation (OLTx). The case of a patient with recurrent disease more than 5 years after OLTx for BCS due to paroxysmal nocturnal hemoglobinuria is analyzed with complete documentation. The literature is reviewed and the probable underlying causes for recurrent disease after OLTx for BCS are discussed including therapeutic consequences. PMID- 10370631 TI - Sialyl Lewis X expression in vascular permeating lesions as a factor for predicting colorectal cancer metastasis. AB - BACKGROUND/AIMS: To evaluate the potential of sialyl Lewis X (SLX) expression for predicting residual liver recurrence, we examined the vascular permeating lesions surrounding the metastatic liver tumors for SLX as a marker of residual liver recurrence. METHODOLOGY: Twenty-eight cases of metastatic liver tumors of colorectal cancers were examined using SLX monoclonal antibody for SLX expression in surrounding vascular permeating lesions. RESULTS: The cumulative residual liver recurrence rate in cases positive for SLX (SLX(+)) expression in the vascular permeating lesions surrounding the metastatic liver tumor was significantly higher than in the negative cases (p < 0.05). Furthermore, logistic regression analysis revealed that SLX positivity in the lesions surrounding the metastatic liver tumor was an important discriminant for residual liver recurrence (p = 0.0197). CONCLUSIONS: These results indicated that SLX expression in vascular permeating lesions surrounding the metastatic liver tumor is a predictive factor for residual liver recurrence. PMID- 10370632 TI - Clinicopathologic and carcinogenetic appraisal of DNA replication error in sporadic T3N0M0 stage colorectal cancer after curative resection. AB - BACKGROUND/AIMS: DNA replication error (RER) was found to play a role in the carcinogenesis of a subset of sporadic colorectal cancers. This study was conducted to evaluate the clinicopathologic implications of RER in T3N0M0 stage colorectal cancers. To better understand the carcinogenesis of sporadic colorectal cancer, the RER status was further correlated with the alterations of K-ras, p53 and deleted in colorectal cancer (DCC) genes which were frequently involved in the adenoma-carcinoma sequence. METHODOLOGY: Seventy-eight patients with curatively resected T3N0M0 stage sporadic colorectal cancer were accumulated. The stored frozen tissues were retrieved for analyses of 1) microsatellite instability at 7 distinct chromosomal loci, 2) loss of heterozygosity at DCC gene, 3) K-ras gene mutation, 4) p53 expression, and 5) DNA content. The RER status was correlated with various clinicopathologic and molecular genetic factors. The survival of patients stratified by RER status was analyzed by Kaplan-Meier estimator. RESULTS: The RER-positive tumor was detected in 32.1% (25/78) of patients. The RER-positive cancer patients presented with distinct clinicopathologic features including young age of tumor onset, proximal tumor location, mucin production in histology, a higher rate of synchronous and metachronous colorectal cancers, and an increased incidence of extracolonic primary cancer. Patients with RER-positive tumor were found to have an improved prognosis with the 5-year survival probability of 76% and 45% in RER-positive and RER-negative groups, respectively (p < 0.05). The RER-positive tumors tended to have normal p53 expression, DNA diploidy, and a lower DNA index. The rate for the loss of heterozygosity of DCC gene was significantly lower in RER-positive tumors. RER status was not associated with K-ras mutation. CONCLUSIONS: The clinicopathologic features and carcinogenesis of RER-positive sporadic colorectal cancers were considered different from those of RER-negative tumors. The presence of RER may identify a subset of less aggressive tumors with good prognosis in T3N0M0 stage colorectal cancers. PMID- 10370633 TI - Pre-operative staging of recto-sigmoid colon carcinoma by upper gastrointestinal endoscopic ultrasonography. AB - BACKGROUND/AIMS: To assess the accuracy of a new generation endoscopic ultrasonography (EUS)(GF-M200) in pre-operative staging of recto-sigmoid colon carcinoma invasion and lymph node metastasis. METHODOLOGY: Seventy-three patients with biopsy proven colon cancer were included in this study. These comprised 60 patients with rectal carcinoma and 13 patients with sigmoid carcinoma. All patients were pre-operatively examined by EUS. Pathological findings of the depth of tumor invasion and presence of lymph node metastasis were correlated with EUS. RESULTS: EUS has an overall accuracy rate of 89% in staging of recto-sigmoid cancer. The diagnostic accuracy rate was 83% for T1, 83% for T2, 93% for T3, and 71% for T4. Understaging and overstaging occurred in 6% and 6%, respectively. In determining lymph node metastasis, the overall accuracy rate was 77%, with a sensitivity and specificity rate of 77% and 76%, respectively. CONCLUSIONS: EUS is a valuable staging modality in the staging of the depth of tumor invasion, not only for rectal carcinoma but also for tumors located at the sigmoid colon. PMID- 10370634 TI - Laparoscopic total proctocolectomy with ileal J pouch-anal anastomosis. AB - BACKGROUND/AIMS: Minimally invasive surgery has developed as one of the most important advances in surgical techniques. The laparoscopic procedure has been successfully used to perform colonic resections. Inflammatory bowel diseases like ulcerative colitis (UC) and familial adenomatous polyposis (FAP) appear as a main indication for total laparoscopic proctocolectomy. METHODOLOGY: At the Second Department of Surgery of the "Regina Elena" Institute for Cancer Research, 5 non selected patients were submitted within a 3-year period (1993-1996) to a total laparoscopic proctocolectomy with a restorative ileal J pouch-anal anastomosis. They comprised 3 males suffering from UC and 2 females affected by FAP. RESULTS: No patients undergoing laparoscopic procedure were converted. The average operative time was 364 min (480 min in the first case, 290 min in the fifth case). There were no intra-operative or post-operative complications (except a mild peritoneal bleeding in the first case, spontaneously stopped). Post operative pain was mild and no analgesics were required. Late results were excellent, with good bowel function within 1 year after the operation, without dietetic, working and sport restrictions and without sexual disorders, mainly in males. CONCLUSIONS: Laparoscopic total proctocolectomy in the hands of skilled laparoscopic surgeons is a feasible, safe and effective procedure, with early and late results comparable to, and in some aspects better than, those obtained with "open" surgery. Moreover, it does not have the disadvantage of intra-operative fluid loss, prolonged post-operative ileus, pain and, in younger patients, psychological discomfort of the wide scar. PMID- 10370635 TI - Colorectal carcinoma: laparoscopic versus traditional open surgery. A clinical trial. AB - BACKGROUND/AIMS: The purpose of this perspective study was to define the role of laparoscopic surgery in the treatment of colorectal carcinoma. METHODOLOGY: One hundred colorectal cancer patients were submitted to surgical treatment between 1993 and 1996. Fifty patients were operated on by videolaparoscopy, the other 50 were operated on according to the standard "open" technique. The two groups had similar demographic (age, gender), pathological (site, stage), and surgical (type and extent of resection) data. Early and late results, benefits and drawbacks of the minimally invasive technique are compared to those of standard open surgery. RESULTS: No intra-operative complications and no operative mortality occurred in the two groups. Early results (complications within 30 days from surgery) were: 1 pneumonia, 3 wound sepsis, and 3 fistulas (one required a reoperation) in the laparoscopic group; 2 wound sepsis and 5 fistulas (spontaneously recovered) in the open group. Late complications occurred in the laparoscopic group only: 1 bowel bridle occlusion 2 months after surgery (that required a reoperation), and 2 stenoses of the colorectal Knight-Griffen anastomosis, successfully treated by dilatation. Concerning the oncologic results, data were calculated on 40 laparoscopic and 43 open curative resections (stage I, II and III): 20% (8/40) of the laparoscopic and 23% (10/43) of the open group patients resulted in neoplastic progression. The neoplastic recurrences were single site (liver or regional) in 3 laparoscopic and in 5 open patients; multiple sites of relapse were observed in 5 laparoscopic (liver, peritoneum and 1 trocar site) and in 5 open (liver, peritoneum and 1 scar) cases. Five-year disease-free survival rates (Kaplan-Meier method) were similar in the two groups: 73.2% in the laparoscopic and 70.1% in the open. CONCLUSIONS: Laparoscopic surgery seems to be a feasible and effective treatment of colorectal cancer and, with the improvement of technology and surgeon skill, it will represent an excellent alternative to the more diffuse and consolidated open surgery technique. PMID- 10370636 TI - CEA and CA 19-9 as prognostic indexes in colorectal cancer. AB - Carcinoembryonic Antigen (CEA) and CA 19-9 are tumor markers expressed by colorectal cancers (CR), particularly in advanced cases. The aim of this study was to evaluate the prognostic value of pre-operative elevated CEA and/or CA 19-9 levels for patients with CR. Blood samples were collected from 74 patients. CEA and CA 19-9 were determined by ELISA (normal range: 0-3 ng/ml for CEA and 0-37 U/ml for CA 19-9). All patients were followed-up for at least 30 months or until death. At the time of diagnosis, 42% of the patients had elevated serum levels of CEA and 35% of CA 19-9. Relapse was observed in 33 patients, 73% of whom had elevated CEA and/or CA 19-9 levels. Among patients without relapse, 68% and 73% had normal values of CEA and CA 19-9, respectively. Ninety-three percent of patients, who had CR recurrence during the first year, had an elevated CEA and/or CA 19-9 level, while 67% of the patients with CR after 1 year, had normal tumor markers. Elevated pre-operative serum CEA and CA 19-9 levels were each predictive of increased cancer mortality (p = 0.001 for CEA, p = 0.01 for CA 19-9). Raised CEA and CA 19-9 levels identify patients at high risk for CR and death and may be useful in selecting patients for adjuvant therapy. PMID- 10370637 TI - A minimally invasive approach to rectal cancer--sacrolaparoscopic approach. AB - BACKGROUND/AIMS: To investigate the feasibility of a technique with minimal skin incision, while retaining a rate of cure and safety equivalent to conventional rectal amputation, by making use of the advantages of laparoscopic procedures, we performed a minimally invasive laparoscopic rectal amputation. METHODOLOGY: Six patients suffering from rectal cancer with cardiac and/or respiratory disorders underwent laparoscope-assisted rectal amputation. The procedure was performed in three steps: 1) sacral approach, 2) laparoscope-assisted abdominal approach under CO2 insufflation, and 3) extracorporeal resection of the inferior mesenteric artery (IMA) and stoma making without CO2 insufflation. RESULTS: Intra-operative cardiopulmonary functions were maintained within normal range during CO2 insufflation. Although all patients had severe respiratory or cardiac disorder or diabetes mellitus, no complications were observed during and after surgery. The post-operative course was uneventful for our patients, each of whom could eat on the first post-operative day and walk on the third post-operative day. All patients were discharged from the hospital uneventfully. CONCLUSIONS: Laparoscope assisted rectal amputation is technically feasible, adequate tumor excision can be achieved with it and post-operative recovery is improved. Sacrolaparoscopic rectal amputation appears to be a safe alternative procedure for patients with rectal cancer and even with severe cardiopulmonary disorders. PMID- 10370638 TI - Serological studies on CEA, CA 19-9, STn and SLX in colorectal cancer. AB - BACKGROUND/AIMS: Sialyl Le(x) (NeuAca2-3Galb1-4(Fuca1-3)GlcNAc1-R) (SLX) was introduced as cancer-associated. In this study, serological expression of SLX was examined with that of carcinoembryonic antigen (CEA), carbohydrate antigen 19-9 (CA 19-9) and sialyl Tn antigen (STn) concerning the prognosis and clinicopathological findings of patients with colorectal carcinoma. METHODOLOGY: One hundred and seventeen patients were operated on for colorectal cancer and were enrolled in this study. Serum CEA, CA 19-9, STn and SLX levels were measured pre-operatively by radioimmunoassays and the cut-off values were 5ng/ml, 37U/ml, 45U/ml, and 38U/ml, respectively. RESULTS: Serologically, the positive rates of CEA, CA 19-9, STn and SLX antigens were 33.3, 26.5, 23.1, and 17.1%, respectively. The CEA, CA 19-9 and SLX are stage specific in clinical stage. In the CEA, CA 19-9, STn, SLX-positive patients, 5-year survival rates were 41.0, 29.7, 50.1, and 29.1%, respectively. In the tumor marker-positive patients, prognosis was significantly poorer than in the negative patients. In the patients with Curability A or B, the difference in survival between the SLX-positive and negative patients was significant. Multivariate analysis revealed that significant prognostic indicators were accompanying liver metastasis, histological type, depth of invasion, CEA and SLX. CONCLUSIONS: The combination assay of serum CEA, CA 19-9, STn and SLX will be beneficial for diagnosis and follow-up of colorectal cancer. PMID- 10370639 TI - Increased production of vascular endothelial growth factor by intestinal mucosa of patients with inflammatory bowel disease. AB - BACKGROUND/AIMS: Vascular endothelial growth factor (VEGF) is a heparin-binding glycoprotein with potent angiogenic, mitogenic and vascular permeability enhancing activities specific for endothelial cells. Recent studies have shown significantly increased VEGF serum levels in patients with active Crohn's disease and ulcerative colitis. The origin of the circulating VEGF is not yet completely described. The present investigation examines the VEGF production of colonic mucosa in consideration of mucosal disease activity in patients with inflammatory bowel disease. METHODOLOGY: Fifteen patients with inflammatory bowel disease were studied, 9 patients with Crohn's disease and 6 patients with ulcerative colitis. Biopsies were taken from endoscopically inflamed and non-inflamed colonic mucosa. Therefore, an analysis of the spontaneous VEGF production of cultured biopsies without stimulus and of the histological grade of inflammation scored on a scale of 0-3 (normal mucosa--severe chronic colitis) were performed. Eight patients with irritable bowel syndrome served as controls. VEGF levels in the supernatant of cultured mucosal biopsies were measured using an enzyme linked immunosorbent assay. RESULTS: VEGF production is expressed as pg/mg wet weight of the biopsies. Inflamed mucosa of patients with active ulcerative colitis (16.27 +/- 10.39, p = 0.003, n = 6) and active Crohn's disease (9.88 +/- 5.98, p < 0.012, n = 9) showed a significantly higher spontaneous production of VEGF by colonic mucosa than normal mucosa of controls (3.16 +/- 1.63, n = 8). In addition, there was an increased unstimulated VEGF production by cultured inflamed mucosa of patients with Crohn's disease compared with non-inflamed mucosa (3.88 +/- 3.66, p < 0.015, n = 9). In both Crohn's disease and ulcerative colitis, there was no significant difference between VEGF production by non-inflamed mucosa and normal mucosa of controls. CONCLUSIONS: The present study identifies the intestinal mucosa as one of the origins of the elevated VEGF serum levels in patients with active inflammatory bowel disease and verifies the findings of recent studies about the importance of VEGF in Crohn's disease and ulcerative colitis. PMID- 10370641 TI - Intramural duodenal hematoma of pancreatic origin. AB - Non-traumatic intramural hematoma of the duodenum is an unusual clinical entity. Indeed, in a majority of 70% of patients intramural hematoma of the duodenum is caused by a blunt, frequently minor abdominal trauma. The main etiology of non traumatic intramural hematoma of the duodenum in the adult is overdose anticoagulant therapy. Rarer causes include pancreatic disease, blood dyscrasia or vascular collagen disease. In this presentation a case of pancreatitis-induced intramural duodenal hematoma is discussed and compared with corresponding data in the literature. PMID- 10370640 TI - Laparoscopic treatment of peptic ulcers. A review after 6 years experience with Hill-Barker's procedure. AB - BACKGROUND/AIMS: This study illustrates our experience in treating duodenal ulcers by means of laparoscopy over a period of 6 years and the results after a lengthy careful follow-up. METHODOLOGY: From October 1991 to October 1997 we submitted 35 patients, 28 men and 7 women of an average age of 51 years (range: 22-78), to vagotomy with minimally invasive access: 23 Hill-Barkers, 2 Taylors, 6 thoracoscopic truncal vagotomies, and 4 laparoscopic truncal vagotomies. Of the patients submitted to surgery with the Hill-Barker technique, 8 were resistant to medical therapy, 11 decided not to continue with long-term medical therapy, 3 assumed an irregular medical therapy, and 1 who had been suffering for a long time from an ulcerous disease required vagotomy in association with laparoscopic cholecystectomy. In 16 patients a bleeding complication preceded surgery. RESULTS: In our experience, the average duration of the operation with the Hill Barker technique is 40 min (range: 30-80 min), with return to normal feeding in 1 day without any disorders and return home on day 3 (range: 2-5). The patients have been followed for 3-54 months. One patient (4.3%) was lost during the follow up. Twenty-one (91.3%) out of the 23 submitted to anterior superselective and posterior truncal vagotomy were pain and ulcer-free without medical therapy. There was only one relapse (4.3%) after 7 months where the patient underwent left thoracoscopic truncal vagotomy. CONCLUSIONS: In our opinion, as posterior truncal and anterior superselective vagotomy using the Hill-Barker technique guarantees the same excellent results, it is preferable due to the speed and ease of performance and to the low cost compared with other procedures which take more time (e.g., Taylor's section and suture of the anterior gastric wall) and require the use of particularly expensive equipment (e.g., Gomez-Ferrer's mechanical sectioning and suturing). PMID- 10370642 TI - Prediction of the lower esophageal sphincter pressure after oral molsidomine by sydnonimine plasma concentrations. AB - BACKGROUND/AIMS: Recent studies suggest that endogenous nitric oxide decreases lower esophageal sphincter pressure (LESP). Substances leading to the formation of nitric oxide, such as molsidomine, decreases the human LESP. It is not yet clear whether this reduction is related to plasma concentrations of molsidomine, the nitrate-active substance sydnonimine (SIN-1) or to serum concentrations of nitrate/nitrite (NOx) as a stable end-product of volatile nitric oxide. METHODOLOGY: We performed a double blind controlled crossover trial in 8 healthy male volunteers. Plasma concentrations of molsidomine, SIN-1 and serum concentrations of NOx as well as esophageal manometry were determined. RESULTS: Mean basal LESP was significantly decreased from 25.4 +/- 2.8 mmHg to 21.9 +/- 2.7 mmHg and 21.4 +/- 2.6 mmHg 2 and 3 hours after molsidomine administration, respectively (mean +/- SEM; n = 8; p < 0.05). The maximum decrease of LESP from the baseline within 1-4 hours after molsidomine administration was 7.6 +/- 1.5 mmHg (mean +/- SEM; n = 8; p < 0.01). The decrease of the LESP correlated significantly with plasma concentrations of SIN-1 (r = -0.53; p = 0.002). NOx levels remained unchanged. CONCLUSIONS: Molsidomine decreases the LESP and plasma concentrations of the active metabolite SIN-1 may predict the potency of molsidomine to lower LESP. NOx was useless as a control metabolite to measure the LESP in response to molsidomine in healthy volunteers. PMID- 10370643 TI - Beta 1 integrin expression in adenocarcinoma of Barrett's esophagus. AB - BACKGROUND/AIMS: In vitro and in vivo studies did not show that beta 1 integrin expression is associated with malignant transformation or that it is of prognostic value in some malignant tumors. There are no data on the expression or prognostic value of beta 1 integrins in adenocarcinoma of Barrett's esophagus. METHODOLOGY: We assessed the expression pattern and the prognostic impact of beta 1 integrins in paraffin-embedded specimens of 41 patients with adenocarcinoma of Barrett's esophagus by immunochemistry. At the time of investigation, neither histomorphological parameters nor the survival time were known. RESULTS: There was no correlation between histomorphological parameters and the expression of beta 1 integrins. The expression of beta 1 integrins had no influence on long- term survival. There was a relationship between the prognosis and the following histopathological parameters: pT, pN and pM category, the UICC stage, the presence of lymphangiosis, and the DNA content of the tumor cells. CONCLUSIONS: The preliminary results obtained in this study did not show that the expression of beta 1 integrins was of prognostic value in patients with adenocarcinoma of Barrett's esophagus. Further studies in a larger number of patients are required to confirm the results obtained in this investigation. PMID- 10370644 TI - Immunohistochemical expression of thymidine phosphorylase/platelet-derived endothelial cell growth factor in squamous cell carcinoma of the esophagus. AB - BACKGROUND/AIMS: Thymidine phosphorylase (dThdPase) is identical to platelet derived endothelial cell growth factor which is an angiogenic factor. We attempted to clarify the significance of dThdPase expression in esophageal squamous cell carcinoma (SCC). METHODOLOGY: Tissues samples were taken from 50 patients with esophageal SCC after curative surgery. The expression of dThdPase was immunohistochemically examined using a monoclonal antibody to dThdPase (clone 654-1). Microvessels in SCC stained for Factor VIII-related antigen were counted. Vascular endothelial growth factor (VEGF) was immunostained with R11. Ki-67 antigen was immunostained with MIB-1, terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate biotin nick end labeling was performed, and Ki 67 labeling index (LI) and apoptotic index were calculated. RESULTS: The expression of dThdPase was observed in 30 patients (60%). Between the SCC with and without dThdPase expression, significant differences were found in microvessel count (p < 0.001) and VEGF expression (p < 0.01), but not in Ki-67 LI and apoptotic index. With regard to the clinicopathologic factors, significant differences were observed in histologic venous invasion (p < 0.01) and lymph node metastasis (p < 0.05). Survival rate after surgery was better in the patients with dThdPase-negative SCC (p < 0.05), and distant organ metastasis after surgery was frequently observed in the patients with dThdPase-positive SCC (p < 0.05). CONCLUSIONS: These results suggest that dThdPase expression may be associated with angiogenic promotion and may be one of the prognostic factors in esophageal SCC. PMID- 10370645 TI - Histochemical study of vascular endothelial growth factor in squamous cell carcinoma of the esophagus. AB - BACKGROUND/AIMS: Vascular endothelial growth factor (VEGF) plays a key role in tumor angiogenesis. The aim of this study was to clarify the significance of VEGF expression in esophageal squamous cell carcinoma (SCC). METHODOLOGY: Tissues samples were taken from 52 patients with esophageal SCC after surgery. VEGF expression in these SCCs was examined immunohistochemically. Microvessels in the tumor stained for Factor VIII-related antigen were counted. Ki-67 antigen as a proliferative marker was immunostained with MIB-1, and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate biotin nick end labeling was performed for the evaluation of apoptosis. Ki-67 labeling index (LI) and apoptotic index were then calculated. RESULTS: VEGF expression was observed in 30 of the patients (57.7%). The microvessel count was significantly higher (p = 0.007), and the apoptotic index was significantly lower (p < 0.0001) in the SCC with VEGF expression than in the SCC without it, but no significant difference was observed in the Ki-67 LI between these groups. There was an inverse correlation between the microvessel count and the apoptotic index (p = 0.007). In the clinicopathologic factors, histologic venous invasion of cancer cells (p = 0.039) and lymph node metastasis (p = 0.049) were significantly correlated with VEGF expression. The survival rate after curative surgery was better in the patients without VEGF expression (p < 0.05), and distant organ metastasis after surgery was frequently observed in the patients with VEGF expression (p = 0.023). CONCLUSIONS: These results suggest that VEGF expression is associated with angiogenesis in esophageal SCC, and may be a prognostic factor in patients with esophageal SCC. Furthermore, apoptosis may be influenced by angiogenesis in esophageal SCC. PMID- 10370646 TI - Endoscopic balloon dilation for benign esophageal anastomotic stricture: factors influencing its effectiveness. AB - BACKGROUND/AIMS: The aim of this study was to identify factors that might affect the results of treating benign anastomotic stricture of the esophagus with balloon dilation. METHODOLOGY: Balloon dilation was performed on 35 patients with benign esophageal anastomotic stricture of the upper (esophageal cancer: 18) or lower (gastric cancer: 15, esophageal varices: 2) esophagus. The procedure was considered effective when patients were able to maintain a solid diet more than 12 months after the last dilation. The follow-up period ranged from 15-130 months (mean: 51 months). RESULTS: A total of 245 dilations were performed, with an average of 6.6 dilations per patient. Treatment was effective in 29 patients (83%). Balloon dilation was successful when treating strictures shorter than 12 mm in length. The strictures were significantly shorter in patients treated effectively (5.6 vs. 30.8 mm). The diameter of the stricture did not affect the results. All the strictures in the lower esophagus and all those resulting from stapled anastomoses were treated successfully, while the effectiveness of treating strictures in the upper esophagus or those resulting from hand-sewn anastomoses was 67% and 57%, respectively. Strictures without prior leakage were treated effectively 92% of the time, while the success rate fell to 56% if there was a preceding leak. An average of 4.4 dilations were performed in effective cases, while the average was 17.5 dilations in ineffective cases. The number of repeat dilations was correlated with the length of the stricture. CONCLUSIONS: Balloon dilation can successfully treat strictures shorter than 12 mm long. The correlation equation may be used to predict the number of repeat dilations and treatment results, and is useful for deciding when to use an alternative method to balloon dilation. PMID- 10370647 TI - Endoscopic management for bleeding esophageal varices: sclerotherapy versus sclerotherapy plus band ligation versus band ligation alone. One year experience at a main hospital in Saudi Arabia. AB - BACKGROUND/AIMS: This study was done retrospectively to compare the outcome of sclerotherapy alone, band ligation alone and band ligation alternating with sclerotherapy in treatment of esophageal varices. METHODOLOGY: During 1 year 30 patients were admitted with variceal bleeding. They received either injection sclerotherapy (8 patients) or band ligation (11 patients), and 11 patients had a combination of both either during first bleed or during follow-up therapy, which is more than 2 sessions in each group. RESULTS: The success rate for stopping first bleeding was 100% in the band ligation and sclerotherapy alone group. The rebleeding rate was 27% in the combination group, 9% in the band ligation group, and none had rebleeding in the sclerotherapy group during follow-up. Eradication of varices was observed in 33% of patients after a second set of sclerotherapy and band ligation. CONCLUSIONS: Our study showed no significant difference between sclerotherapy versus band ligation in stopping initial bleeding or eradication of varices during the follow-up period, but there was a difference in re-bleeding rates among the three groups. PMID- 10370649 TI - Possibility of pre-operative diagnosis of lymph node metastasis based on morphology. AB - BACKGROUND/AIMS: The accuracy of pre-operative diagnosis of lymph node metastasis is insufficient. Our aim was to define the possibility of diagnosing metastatic lymph nodes based on morphology. METHODOLOGY: One hundred and fifty-seven patients with pre-operatively untreated esophageal squamous cell carcinoma underwent resection, 5334 lymph nodes were isolated, and the short and long diameters were measured. We tried to construct a linear regression line for metastasis rate versus lymph node size (long diameter classified at intervals of 1 mm) by each location. The ratio of short diameter to long diameter (SL ratio) of metastasis-positive lymph nodes was compared with that of negative ones at each location. RESULTS: Gradient and intercept of overall regression line was 0.0213 and 0.0101, respectively, and the long diameter producing a metastasis rate of 80% (LD80) was 37.1 mm. Metastasis-positive lymph nodes larger than calculated LD80 represented no more than 9.5% of all the corresponding metastasis positive nodes. The locations with significant difference of SL ratio between metastasis-positive and negative ones were limited to right cardiac, left gastric artery, thoracic paratracheal, bifurcation, and the highest mediastinal nodes. CONCLUSIONS: There is a low possibility that lymph node metastasis can be exactly diagnosed pre-operatively based on the size and morphology. PMID- 10370648 TI - Palliative treatment of esophageal cancer: self-expanding metal stents versus Postlethwait technique. AB - BACKGROUND/AIMS: The development of new techniques for palliation of esophageal carcinoma with lower morbidity and mortality than surgical procedures. METHODOLOGY: Between 1981 and 1994, 258 patients with esophageal and cardiac cancer were treated in our Department. We selected two groups: Group A, 25 patients underwent a by-pass with an isoperistaltic gastric tubular (Postlethwait technique) and, group B, in 30 patients we placed 35 autoexpandable esophageal stents. We subsequently performed a retrospective study. RESULTS: In group A, dysphagia was not relieved in 6 patients (24%) and we found no complications in 18 patients (72%). The hospitalization period ranged from 18-50 days. Hospital mortality rate was 24% (6 patients). Mean survival was 5.4 months (range: 3-9 months). All patients in group B, except for 2, were relieved of dysphagia. Two patients (6.6%) died in the immediate post-intubation period though none of the deaths were related to technical complications. Hospitalization period ranged from 5-12 days. Mean survival was 6 months (range: 12 days to 9 months). CONCLUSIONS: Currently, surgical by-pass procedures are restricted to the patient with an incurable disease not identified until operation time. PMID- 10370650 TI - Metastasis of an esophageal carcinoma to a giant gastric ulcer. AB - In patients with esophageal carcinoma it is considered that stomach metastasis is induced mainly via the lymphatic route rather than via the bloodstream route that is common in other types of distant organ metastasis. A 56 year-old patient is reported who underwent synchronous subtotal esophagectomy and total gastrectomy for a middle third esophageal carcinoma and a giant peptic ulcer within the gastric fundus. The final histopathologic examination revealed a squamous cell carcinoma of the esophagus with concomitant squamous tumor implantation within the gastric ulcer. The increased cell proliferation in the ulcer margin can serve as a "biological background or base" for implantation. PMID- 10370651 TI - ELISA determination of serum hyocholic acid concentrations in humans and their possible clinical significance. AB - BACKGROUND/AIMS: Hyocholic acid (HCA), a bile acid isolated from pigs, has a different structure from the predominant bile acids from humans. METHODOLOGY: We prepared an antiserum to HCA in rabbits and developed an enzyme-linked immunosorbent assay (ELISA), which we used to measure serum HCA in healthy subjects and patients with a variety of gastrointestinal and non-gastrointestinal diseases. RESULTS: Patients with hepatic cirrhosis had a mean HCA concentration that was 120-fold greater than that in healthy subjects. Markedly elevated HCA levels were also present in patients with primary hepatoma or pancreatic cancer but not in patients with cancer of the breast, bile duct, duodenum, or stomach. CONCLUSIONS: If these results are confirmed by further study, HCA measurement may prove clinically useful. PMID- 10370652 TI - Quantification of thymidylate synthase gene expression in human gastrointestinal carcinoma tissues using competitive PCR. AB - BACKGROUND/AIMS: Thymidylate synthase (TS) is a target enzyme for 5-fluorouracil (5-FU). It is important to know the TS expression level in tumor tissue for chemotherapy. We evaluated the TS expression level using competitive polymerase chain reaction (cPCR), and estimated the reliability and reproducibility of this method. METHODOLOGY: TS expression was assessed by both Fluorodeoxyuridine monophosphate (FdUMP) binding assay and cPCR in 9 human adenocarcinoma cell lines and 50 human adenocarcinoma tissues obtained by surgical resection. TS expression was also assessed by cPCR in nine biopsy specimens obtained before surgery. We synthesized TS and beta-actin competitors. Following cPCR, PCR products were quantified on ethidium bromide-stained gels using a digital image analyzer. TS mRNA/beta-actin mRNA ratio was used to determine relative TS expression level. RESULTS: The number of FdUMP binding sites on TS and TS mRNA/beta-actin mRNA ratio were significantly correlated in both cell lines and surgically resected specimens (p < 0.01). A linear regression line formed from the data points was obtained for TS mRNA/beta-actin mRNA ratios in surgical specimens versus TS mRNA/beta-actin mRNA ratios in biopsy specimens (p < 0.01). CONCLUSIONS: Assessment of TS expression by cPCR using our competitors was accurate and reproducible. PMID- 10370653 TI - Gastroduodenal lesions in children with juvenile rheumatoid arthritis. AB - BACKGROUND/AIMS: There are few studies about gastrointestinal abnormalities in patients with juvenile rheumatoid arthritis-probably due to the fact that this association is not frequently recognized. The aim of our study was to observe the prevalence of endoscopic gastroduodenal lesions in these patients. METHODOLOGY: Fourteen patients with juvenile rheumatoid arthritis, all of them using non steroidal anti-inflammatory drugs associated or not with methotrexate, were assessed clinically and by endoscopy. Gastric antrum biopsy and Helicobacter pylori search were also performed. RESULTS: The mean age of the patients was 10.6 years (7 boys). Abdominal pain was observed in 27% of them. Macroscopic endoscopic lesions were found in 43% and infection by Helicobacter pylori in 57%. The correlation between anemia and endoscopic abnormalities was statistically significant (p < 0.05). CONCLUSIONS: Our data show that patients with juvenile rheumatoid arthritis have considerable susceptibility to gastroduodenal lesions, especially if they are using any drug association and present anemia. PMID- 10370654 TI - Endoscopic incision and balloon dilatation for cicatricial anastomotic strictures. AB - BACKGROUND/AIMS: Endoscopic incision or balloon dilatation is common therapy for cicatricial anastomotic strictures after gastrointestinal surgery. These therapies are not always effective. METHODOLOGY: There were 6 patients who failed either endoscopic incision or balloon dilatation alone and who underwent a combination of the two therapies. Two or three small radial incisions were made in the scar of the stricture with the endoscopic electrocautery under direct vision with fiberscopy. Then, the incisions were split bluntly and the stenosis was dilated over 15-20 minutes with balloon-dilator. This procedure was performed once or twice at a 2-week interval. RESULTS: In 5 of the 6 patients, the stenosis was improved in subjective criteria and objective symptoms. In the last patient, only objective improvement was noted. There were no complications. CONCLUSIONS: Endoscopic incision plus balloon dilatation is an effective and safe treatment for cicatricial anastomotic strictures which have failed either therapy alone. PMID- 10370655 TI - Effect of non-steroidal anti-inflammatory drugs on gastric and duodenal prostaglandin concentrations in patients with Helicobacter pylori infection. AB - BACKGROUND/AIMS: Both Helicobacter pylori and non-steroidal anti-inflammatory drugs are reported to affect gastroduodenal prostaglandin synthesis. However, their influence on gastric mucosal prostaglandins remains unclear. The aim of this study was to investigate the influence of nonsteroidal anti-inflammatory drugs on mucosal prostaglandin synthesis in patients with Helicobacter pylori infection. METHODOLOGY: We enrolled 87 Helicobacter pylori-infected patients in this study (gastric ulcer: 33, duodenal ulcer: 41, and non-ulcer dyspepsia: 13). Of them, 27 patients received non-steroidal anti-inflammatory drugs. Endoscopy was performed and biospy specimens from gastric body, antrum and duodenal bulb were assessed for Helicobacter pylori and prostaglandin concentration. RESULTS: A significantly lower mucosal prostaglandin E2 level at gastric body (142.2 +/- 28.1 ng/mg vs. 222.0 +/- 12.4 ng/mg, mean +/- SEM) and antrum (131.3 +/- 26.4 ng/mg vs. 226.0 +/- 19.0 ng/mg) was noted in Helicobacter pylori-infected gastric ulcer patients with non-steroidal anti-inflammatory drugs ingestion than in that of patients without non-steroidal anti-inflammatory drugs ingestion (p < 0.05). Using a multivariate analysis, we found that non-steroidal anti-inflammatory drug was an independent variable affecting gastric and duodenal mucosal prostaglandin E2 synthesis in patients with Helicobacter pylori-infected gastric ulcer. CONCLUSIONS: Non-steroidal anti-inflammatory drugs decrease gastroduodenal mucosal prostaglandin E2 synthesis in gastric ulcer patients with Helicobacter pylori infection. PMID- 10370656 TI - Endoscopic placement of a long intestinal tube-device in order to reduce the distress of patients with intestinal obstruction. AB - We have designed the newly devised endoscopic procedure for inserting and placing the intestinal tube into the duodenum in order to reduce the distress of patients with intestinal obstruction. PMID- 10370657 TI - Intra-operative enteroscopy for obscure gastrointestinal bleeding. AB - Small bowel enteroscopy has been reported useful in the non-surgical evaluation of the small intestine in patients with obscure gastrointestinal bleeding but findings may be limited due to incomplete small bowel intubation and a lack of tip deflection. Intra-operative enteroscopy (IOE) is accepted as the ultimate diagnostic procedure for complete evaluation of the small bowel in these patients. Two patients with obscure gastrointestinal bleeding and deep anemia underwent IOE during surgical exploration. Angiodysplastic lesion with a diameter of 3 cm was found at jejunum in the first patient and segmental jejunal resection was performed. Enteroscopy showed red punctate lesions with a diameter of 1-3 mm located at proximal jejunum and extending to the ileum in the second patient. Total jejunal resection was performed. There was no recurrence of gastrointestinal bleeding during 36 months follow-up. PMID- 10370658 TI - Ileus with incarceration of Spigelian hernia. AB - An 85 year-old female with ileus due to incarceration of Spigelian hernia, which was diagnosed pre-operatively with the findings of the abdominal computed tomography and plain radiographic pictures, is reported. A simple hernioplasty was made by suturing the internal oblique and transverse muscles to the rectus sheath. The post-operative condition is satisfactory without any recurrence of hernia. PMID- 10370659 TI - Carcinoid tumor of the ileum with intestinal obstruction. AB - An 83 year-old female presenting with intestinal obstruction due to a carcinoid tumor of the small intestine is herein reported. The intra-operative findings revealed a stenotic lesion and ischemic changes of the ileum. A segmental jejunotomy was performed and a submucosal tumor was recognized as a causal lesion. Histopathological investigation demonstrated the features of carcinoid tumor. PMID- 10370660 TI - Radiological manifestations of liver and gastrointestinal parasitic infections. AB - There is a wide variation in the prevalence of parasitic diseases in different countries, particularly in the Tropics. The clinical status of affected patients varies from normal to severe ill health. Radiology has acquired a major role in the diagnosis and, in some instances, the management of a significant variety of parasitic infections. Plain films, ultrasound, computed tomography and magnetic resonance imaging, together with the help of clinical and laboratory tests, can reach a diagnosis with a high percentage of accuracy. PMID- 10370661 TI - Helicobacter pylori and gastroesophageal reflux disease: friends or foes? AB - Gastroesophageal reflux disease (GERD) is responsible for a high proportion of digestive symptoms attributable to the upper gastrointestinal tract. Helicobacter pylori (H. pylori) is the main etiologic factor in chronic gastritis and gastroduodenal ulcer disease, but its relation with GERD has not yet been established. The aim of this paper is to review the relationship between H. pylori and GERD, trying to answer the question whether a nexus of "friendship" or "hate" exists between them. Although H. pylori may, in theory, represent a cause for GERD, available data suggest that the infection is not a risk factor for the development of GERD, and the microorganism could even represent a protective factor against this disease. The antisecretory effect of proton pump inhibitors (PPIs) seems to depend on the presence of the infection and H. pylori eradication has, therefore, negative consequences on the efficacy of antisecretory drugs (although its possible clinical relevance, precisely in patients with GERD, remains unknown). Moreover, H. pylori eradication in patients with duodenal ulcer disease is associated in some studies, but not in others, with a higher incidence of GERD, although the reported reflux esophagitis is usually mild. It can be concluded, from these data, that investigating or treating H. pylori infection is not recommended in patients with GERD (when these patients do not need PPI maintenance therapy). Finally, it has recently been recommended to eradicate H. pylori infection in those patients with GERD needing long-term treatment with omeprazole, as some studies have reported that this drug induces, in presence of the microorganism, an atrophic gastritis, with the consequent theoretic risk of gastric cancer. However, several arguments against this attitude can be postulated, and noteworthy are the following: many studies suffer important methodological defects, several authors report contrary results, and the possibility that H. pylori could play, as previously mentioned, a protective role against GERD. It may be concluded, therefore, that the indication of eradicating H. pylori in patients with GERD and maintenance therapy with PPIs, although supported by several arguments, cannot be considered as definitively established. In conclusion, H. pylori and GERD seem to have, in any case, a "friendly" relationship, although it may be transformed into one of "hate" when PPIs enter the scene. PMID- 10370662 TI - Correlation between human hepatocyte growth factor and interleukin-6 concentrations after surgery. AB - BACKGROUND/AIMS: The aim of this study was to determine whether the production of human hepatocyte growth factor (hHGF) occurs in injured tissue following surgery in conjunction with interleukin-6 (IL-6) synthesis, and independently of liver resection. METHODOLOGY: The post-operative concentrations of hHGF and IL-6 in the serum and local exudative fluid were measured in 33 patients following abdominal (n = 20) or thoracic surgery (n = 13) (non-HTX), and in 7 undergoing a partial hepatectomy (HTX). RESULTS: Concentrations of two cytokines in the fluid were much higher than those in the serum (p < 0.001) in all patients. The maximum serum levels of hHGF were significantly correlated with those of IL-6 (p < 0.05), intra-operative blood loss (p < 0.005), and operating time (p < 0.005) in patients undergoing non-HTX. In these patients, the fluid levels of hHGF were also significantly correlated with those of IL-6 (p < 0.001). In serum and fluid, the hHGF/IL-6 ratios were markedly higher after HTX than after non-HTX (p < 0.05). CONCLUSIONS: These results suggests that hHGF may be produced in conjunction with IL-6 in injured tissue following surgery, but the production of hHGF may predominate over that of IL-6 in the liver following partial hepatectomy. PMID- 10370663 TI - Surgical management for adrenal gland metastasis of hepatocellular carcinoma. AB - BACKGROUND/AIMS: Although the adrenal gland is one of the common sites for metastasis from hepatocellular carcinoma, the significance of adrenalectomy for treatment of metastatic hepatocellular carcinoma still remains unclear. METHODOLOGY: Analysis of 4 patients with adrenal metastasis from among 390 patients with hepatocellular carcinoma admitted to our department between October 1994 and December 1997, and a review of 79 cases reported between 1984 and 1997, were performed. RESULTS: Four patients with adrenal metastasis, right in 2 and left in 2, underwent surgical treatment. Three of the patients developed tumor thrombi in the renal vein or inferior vena cava. Diagnosis of adrenal metastasis was made by ultrasonography or computed tomography scan, following an increase in serum tumor markers. Adrenalectomy with removal of the venous tumor thrombi was performed successfully. Three patients died of recurrence within 1 year, and one patient died due to other causes showing no recurrence at autopsy. The literature review revealed the left-sided metastases were significantly larger than the right-sided ones (p < 0.01). There was no significant difference in the survival periods between left and right metastasis, or between patients who underwent adrenalectomy and those treated by other means (p > 0.05). CONCLUSIONS: The rationality of surgical treatment for adrenal metastasis from hepatocellular carcinoma still remains controversial. However, we believe that adrenalectomy would be a safe procedure and increases the chance of survival for patients. PMID- 10370665 TI - Resection of peritoneal metastases from hepatocellular carcinoma. AB - BACKGROUND/AIMS: To evaluate whether resection of peritoneal metastases arising from hepatocellular carcinoma (HCC) has a role to play in the management of the disease. METHODOLOGY: Resections of peritoneal metastases from HCC were performed in 6 patients. The survival of the patients was evaluated in relation to feature of primary liver tumor, number of peritoneal metastases and period between hepatectomy and resection of peritoneal metastases. RESULTS: Two patients had peritoneal metastases at the time of hepatectomy for HCC, their resection being carried out synchronously. In the other 4 patients, peritoneal metastases became evident between 6 and 34 months (mean: 18) after hepatectomy; resection was performed at the time of presentation of the metastases. Patient survival after resection of the peritoneal metastases ranged from 3-31 months. The 4 patients who survived for more than 1 year had the following features: 1) a small number of metastatic nodules (= or < 4); 2) low alpha-fetoprotein (AFP) values (mean: 205 ng/ml); and, (3) metachronous occurrence of the peritoneal metastases. CONCLUSIONS: Resection of peritoneal metastases arising from HCC may be of value in improving patient survival. PMID- 10370664 TI - Adjuvant hepatic arterial infusion chemotherapy after radical hepatectomy for hepatocellular carcinoma--results of long-term follow-up. AB - BACKGROUND/AIMS: This clinical study aimed to clarify the effectiveness and indication of adjuvant hepatic arterial infusion chemotherapy (HAIC) that is performed to prevent recurrence after radical hepatectomy for hepatocellular carcinoma (HCC). METHODOLOGY: From January 1986 to December 1992, 135 HCC patients, who tolerated curative hepatic resection in which all of the macroscopic HCC was removed, were included in this study. They were divided into two groups. One group was comprised of 68 patients who received HAIC after radical hepatectomy (HAIC (+) group), and the other group was comprised of 67 patients who received radical hepatectomy alone (HAIC (-) group). In the HAIC (+) group, an emulsion of doxorubicin (30-50 mg) and lipiodol (3-5 ml) was injected from a reservoir every 2 or 3 months for 1 year. RESULTS: The cumulative survival rates in the HAIC (+) group (79.1%, 54.5% and 39.9% at 3, 5, and 7 years after hepatectomy, respectively) were better than those in the HAIC (-) group (69.2%, 38.1% and 26.8%, respectively) (p = 0.086). The disease-free survival rates in the HAIC (+) group (50.8%, 31.7% and 25.6% at 3, 5, and 7 years after hepatectomy, respectively) were significantly better than those in the HAIC (-) group (25.7%, 20.6% and 6.4%, respectively) (p = 0.006). This improvement was evident for 3 years after hepatectomy. The adjuvant HAIC was effective especially in patients with good liver function, whose tumor size ranged between 2.1 cm and 5 cm in diameter, and who received a minor hepatic resection. CONCLUSIONS: Adjuvant HAIC was effective in preventing recurrence after radical hepatectomy for HCC. This treatment is especially indicated for patients with good liver function, whose tumor size ranges between 2.1 cm and 5 cm in diameter, and who have received a minor hepatic resection. PMID- 10370666 TI - TT virus infection in chronic liver disease. AB - BACKGROUND/AIMS: The exact role of the novel hepatotropic TT virus regarding the etiology of viral hepatitis, as well as the progression towards chronic liver disease has as yet not been defined. Moreover, the contribution of TTV infection to the course of chronic hepatitis B or C virus infections also still awaits clarification. Hence, the aim of our study was to investigate the impact of TTV infection on clinical severity and histology of chronic liver disease originating from HBV and/or HCV infections in Thai patients concomitant with the determination of TTV's association with non-B, non-C chronic liver disease and compared to its prevalence among voluntary blood donors. METHODOLOGY: DNA was extracted from the sera collected from 115 hepatitis B patients, 41 hepatitis C, and 48 negative for either viral marker, who had all been diagnosed with chronic liver disease ranging from chronic hepatitis over cirrhosis to hepatocellular carcinoma. The sera obtained from 200 voluntary blood donors served as controls. TTV DNA was amplified by seminested polymerase chain reaction (PCR) employing primers derived from the genome's most conserved region. The PCR products were analyzed by gel electrophoresis. Liver function tests were performed by means of a chemical analyzer. RESULTS: TTV DNA was detected in 20% of the HBV-positive and 19.5% of the HCV-positive chronic liver disease patients. Within the group of patients seronegative for both viral markers, TTV was detected in 8.3%. Furthermore, its DNA was identified in 6.8% of the HCC patients and finally, in 7% of the blood donors. Yet, no significant differences between TTV infected and non-infected patients were found as to demographic data, assumed source of infection, biochemical abnormalities, or severity of liver histology. CONCLUSIONS: TTV appears to be highly prevalent on a worldwide scale but regarding etiology of and progression towards serious liver disease, its contribution seems to be minor if not altogether non-existent. Hence, regarding clarification of its clinical significance, further studies are certainly required. PMID- 10370667 TI - Long-term efficacy of recombinant interferon alpha 2a in the treatment of chronic hepatitis C: a randomized prospective study comparing two dose schedules in Chinese patients. AB - BACKGROUND/AIMS: This study is the first randomized prospective clinical trial of interferon in hepatitis to be conducted according to the guidelines of Good Clinical Practice (GCP) in China. The object of this study is to compare the long term efficacy of a dose of 3MU of recombinant IFN alpha 2a (rIFN-alpha 2a) three times a week (t.i.w.) for 6 months with a starting dose of 6MU for 3 months and subsequent reduction to 3 MU t.i.w for a further 3 months. METHODOLOGY: Sixty eight serological and histologically proven chronic hepatitis C patients with elevated serum ALT were randomized into two groups. A total of 63 patients were studied with full course of treatment. Five patients were withdrawn from the trial, 2 due to personal reasons and 3 due to adverse drug reactions during treatment. Thirty patients received 6MU IFN-alpha 2a t.i.w. for 3 months followed by 3MU t.i.w. for another 3 months (group A). Thirty-three patients received 3MU IFN-alpha 2a t.i.w. for 6 months (group B). RESULTS: The sex, age, baseline serum bilirubin, ALT and AST levels were matched in both groups. At the end of 6 months the complete and partial response rates in group A were 60.0% and 16.7%, respectively, and the clearance of serum HCV-RNA was 53.3%. In group B, the complete and partial response rates were 72.7% and 3.0%, respectively, and the clearance of HCV-RNA was 61.3%. These patients were followed up for 6, 12, and 18 months after stopping treatment. In group A, the rates of complete normalization of ALT and clearance of serum HCV-RNA at 24 months were 50.0% and 60.0%, respectively. In group B, the rates of normalization of ALT and clearance of HCV RNA at 24 months were 54.4% and 41.9%, respectively. The efficacy between the two groups showed no statistically significant difference; the response rates of treatment were similar to the patients with HCV genotype 1b and 2a. Six patients (10.8% of the study population) developed neutralization antibodies to IFN-alpha 2a during treatment, 4 of them responded to the treatment. Adverse drug reactions (ADR) were common, but most of them were tolerable and the incidence of ADR was similar in both groups. CONCLUSIONS: IFN-alpha 2a is effective in the treatment of Chinese patients with chronic hepatitis C. The sustained response rates and ADR among two dose schedule groups are similar. PMID- 10370668 TI - Survival after repeat hepatic resection for recurrent colorectal metastases. AB - BACKGROUND/AIMS: This is a retrospective study examining survival of patients undergoing repeat hepatic resection for recurrent colorectal metastases. METHODOLOGY: The records of 41 patients undergoing hepatic resection for metastatic colorectal cancer were reviewed. Curative resections (negative resection margin and no extrahepatic disease) were attempted in all patients. Recurrence developed in 26 (63%) patients, with disease being confined to the liver in 16 (39%) patients. Ten of them (24%) underwent hepatic resection and make up the study population. RESULTS: Ten patients (4 women, 6 men; mean age: 62 years, range: 50-82 years) developed recurrence confined to the liver at the median interval of 16 months (range: 5-34 months) after the first hepatectomy. In 6 patients the recurrent cancer(s) involved both the area near the resection line and remote sites from the site of the first hepatic resection. In 3 patients recurrent cancer(s) was located at sites remote from the first liver resection. In 1 patient the recurrent cancer was located in the same area as the original hepatic resection. Three formal hepatectomies and seven non-anatomical (wedge) resections were performed. The mean blood loss was 900 cc (range: 100-2700 cc); the mean hospital stay was 19 days (range: 8-34 days). There was no perioperative mortality. Morbidity was 20%. Four patients died of recurrent disease, with a mean disease-free survival of 13 months (range: 5-21 months). Two patients had a second recurrence resected at 10 and 24 months, respectively, after the second hepatic resection. One of these 2 patients had a fourth hepatic resection for hepatic recurrence and is still alive with no evidence of disease. Six patients are alive, 4 of them without evidence of disease, with a median follow-up time of 30 months (range: 22-64 months). Actuarial 4-year specific survival was 44%. Actuarial disease-free survival at 4 years was 18%. CONCLUSIONS: In appropriately selected patients, repeat hepatic resection for colorectal metastases is a worthwhile treatment. Mortality, morbidity, and survival are similar to those following the initial resection. PMID- 10370669 TI - Combined therapy with radiofrequency thermal ablation and intra-arterial infusion chemotherapy for hepatic metastases from colorectal cancer. AB - BACKGROUND/AIMS: In this preliminary study, we investigated the efficacy of combined radiofrequency thermal ablation therapy (RFA) with hepatic arterial infusion chemotherapy (HAI) in the treatment of multiple liver metastases from colorectal cancer. METHODOLOGY: Nine patients with bilobular multiple metastases was treated. The number of nodules was 6.0 +/- 3.9 (range: 2-13), and the size was 2.1 +/- 1.0 cm (range: 0.5-4.8 cm) in diameter. RFA was performed using a RF generator operating at 460 kHz with a 15-gauge, 4-prong custom RF needle. Treatment temperature was kept at 90-110 degrees C for 5 min. 5-Fluorouracil (5 FU) was administered by weekly 750-1250 mg/body/5 h as the regimen of HAI. RESULTS: During a 15.2-month follow-up period, 6 of 9 patients survived more than 1 year. Three of the 6 survived more than 2 years. Serum CEA level in 5 patients dropped from 24.5 +/- 9.5 ng/ml to 10.3 +/- 5.5 ng/ml. Local recurrence was observed in 5 patients and new lesions in 4. Extrahepatic recurrence was observed in 5 patients. There were no serious complications but one HAI-related cerebral thrombosis. CONCLUSIONS: Combined RFA with HAI would be effective and safe. This modality provides a new option for the treatment of multiple liver metastases from colorectal cancer. PMID- 10370670 TI - Portal serum human hepatocyte growth factor levels after partial hepatectomy. AB - BACKGROUND/AIMS: We investigated whether or not hepatocyte growth factor increases in portal serum via an endocrine mode after partial hepatectomy in humans. METHODOLOGY: Portal blood was sampled through a catheter inserted through the umbilical vein to the portal trunk during surgery in 17 patients. Serum human hepatocyte growth factor levels were determined by enzyme-linked immunosorbent assay. RESULTS: Human hepatocyte growth factor levels were higher in portal than in peripheral serum throughout the study. Portal and peripheral serum human hepatocyte growth factor levels without complications increased rapidly and reached a maximum level 1 day after partial hepatectomy. The maximal level of portal and peripheral serum human hepatocyte growth factor was 1.20 and 1.00 ng/ml, respectively. In the case of hepatic failure after partial hepatectomy, portal and peripheral serum human hepatocyte growth factor levels markedly increased and reached 9.31 ng/ml and 6.78 ng/ml 2 days before death, respectively. CONCLUSIONS: These results suggest that hepatocyte growth factor increases in portal serum via an endocrine mode after partial hepatectomy in humans. Furthermore, measurement of the portal and peripheral serum human hepatocyte growth factor levels may be useful for the clinical evaluation of patients with post-operative hepatic failure. PMID- 10370672 TI - Risk factors associated with intra-operative blood loss in hepatectomized patients with giant cavernous hemangioma of the liver. AB - BACKGROUND/AIMS: The aim of this study was to clarify risk factors associated with intra-operative blood loss in hepatectomized patients with giant cavernous hemangioma (GCH) of the liver. METHODOLOGY: Twenty patients with GCH of the liver were treated by hepatectomy. Eleven patients with intra-operative blood loss > 2000 ml (mean: 7145 +/- 7080 m; Group 1) were reviewed retrospectively and compared to 9 patients with intra-operative hemorrhage < 2000 ml (mean: 918 +/- 429 ml; Group 2). RESULTS: Although there were no significant differences in pre operative AST, ALT, and ICG-15 or fibrinogen and platelets between the two groups, pre-operative total bilirubin and fibrin degradation product (FDP) in Group 1 was significantly higher than in Group 2. Mean operation time and intra operative blood transfusion in Group 1 versus Group 2 were 433 min vs. 213 min (p < 0.0001) and 3036 ml vs. 422 ml (p = 0.0072), respectively. The weight of resected liver (r = 0.821, p < 0.0001), maximum diameter of tumor (r = 0.782, p < 0.0001) and operation time (r = 0.748, p < 0.0001) were the most highly correlated with intra-operative blood loss, followed by pre-operative total bilirubin (r = 0.605, p = 0.0038), FDP level (r = 0.576, p = 0.0068) and intra operative blood transfusion (r = 0.561, p = 0.0089). CONCLUSIONS: These findings suggest that pre-operative management to reduce the tumor size, total bilirubin and FDP levels may be essential to minimize intra-operative hemorrhage and blood transfusion. PMID- 10370671 TI - Use of transcatheter arterial infusion of anticancer agents with lipiodol to prevent recurrence of hepatocellular carcinoma after hepatic resection. AB - BACKGROUND/AIMS: Hepatectomy has been accepted as a reliable cure for primary hepatocellular carcinoma (HCC). However, the residual liver recurrence rate after hepatectomy remains high. To improve the prognosis after hepatectomy for HCC, repeated post-operative transcatheter arterial infusions of anticancer drugs and lipiodol (TAI) was given. This study evaluates the efficacy of this treatment for preventing residual liver recurrence after hepatectomy. METHODOLOGY: TAI after hepatectomy was performed in 24 (TAI group) of 65 cases showing tumor invasion such as infiltration to the capsule, intraportal spread, and intrahepatic metastasis. In TAI, a mixture of Mitomycin C (MMC) and Adriamycin (ADM) is administered with lipiodol via the hepatic artery. The recurrence and survival rates of the TAI (n = 24) and non-TAI (n = 41) groups were compared to evaluate the efficacy of TAI after hepatectomy. RESULTS: The TAI group had a lower cumulative residual liver recurrence rate than the non-TAI group (p < 0.01). Division of residual liver recurrence cases into two groups according to the duration of recurrence showed that the rate of recurrence within 1 year after hepatectomy was lower in the TAI group (10.0%) (1/10) than in the non-TAI group (48.4%) (15/31) (p = 0.07). Also, the cumulative survival rate in the TAI group was significantly higher (p < 0.05). The morbidity rate was 16.6%. Bilomas occurred without infection in 2 cases, and liver abscess in one. CONCLUSIONS: TAI may be an effective surgical adjuvant against residual liver recurrence, and we suggest that its effectiveness results from suppression of intrahepatic micrometastases rather than multicentric carcinogenesis. PMID- 10370673 TI - Interferon treatment in patients with chronic hepatitis C with normal alanine aminotransferase activity. AB - BACKGROUND/AIMS: Chronic hepatitis C virus carriers may have repeatedly normal alanine aminotransferase activity despite detectable viremia and histological hepatitis. We aimed to evaluate the effect of interferon treatment in these cases. METHODOLOGY: Twelve patients with persistently normal alanine aminotransferase levels at least 6 months before therapy were treated with recombinant interferon (IFN)alpha-2b for 6 months, totaling 840 MU in amount. Alanine aminotransferase levels were measured monthly during treatment and after treatment withdrawal, and HCV-RNA levels were measured by polymerase chain reaction before treatment, and 6 and 12 months after treatment withdrawal. RESULTS: At treatment withdrawal, HCV-RNA levels had significantly decreased and HCV-RNA disappeared in 9 of the 12 patients by polymerase chain reaction. At 6 months after treatment withdrawal, HCV-RNA reappeared in 6 of the 9 patients whose HCV-RNA was negative at treatment withdrawal. Over all, only 4 of the 12 patients (33%) were sustained virological responders (HCV-RNA is negative more than 6 months after treatment withdrawal). Pre-treatment HCV-RNA levels in a sustained virological responder was significantly lower than that of transient and non-responders (4.9 +/- 1.6 vs. 7.7 +/- 1.6 log10[copies/ml], p < 0.05). Of 8 patients who did not achieved sustained virological response, alanine aminotransferase levels had transiently increased above normal during treatment in one patient and after treatment withdrawal in 6 patients; however, in the remaining one patient abnormal values have continued from 8 months after treatment withdrawal till now for 24 months. CONCLUSIONS: In patients with chronic hepatitis C with normal alanine aminotransferase levels, the response to interferon therapy was by no means satisfactory. However, if it would be used in cases with the lower pre-treatment HCV-RNA levels with careful attention to a transient alanine aminotransferase elevation, the more a sustained virological response might be expected. PMID- 10370674 TI - Relationship between ornithine decarboxylase gene abnormalities and human hepatocarcinogenesis. AB - BACKGROUND/AIMS: Ornithine decarboxylase is essential for cell growth. Its activity was high in human hepatocellular carcinoma tissues and was highest in poorly differentiated tumors. METHODOLOGY: To find if there are tumor-specific ornithine decarboxylases, we examined the ornithine decarboxylase cDNA sequences of 91 clones prepared from hepatoma tissue and non-cancerous tissue of resected liver specimens from 15 patients with hepatocellular carcinoma. RESULTS: Ornithine decarboxylase gene mutations were more frequently detected in the hepatoma tissue. The incidence of mutation in hepatoma tissue was related to dedifferentiation. Mutation in regions rich in proline, glutamic acid, serine, and threonine were detected in moderately and poorly differentiated hepatocellular carcinoma only. CONCLUSIONS: Our findings suggested that the sequence of ornithine decarboxylase in hepatocellular carcinoma often is different from that in normal liver and that mutation of its gene is related to the progression of hepatocellular carcinoma. PMID- 10370675 TI - Homeless patients with hepatocellular carcinoma in Osaka City, Japan. AB - BACKGROUND/AIMS: Although the number of homeless persons is increasing worldwide, studies delineating the health status of these persons according to various medical perspectives, including hepatology, are limited. However, such studies are important for understanding the pathogenesis of diseases and their prevention. METHODOLOGY: Thirty homeless patients with hepatocellular carcinoma (HCC) and 15 with liver cirrhosis (LC) who were admitted to the Osaka Socio Medical center Hospital during the past 6 years were analyzed clinicopathologically. All were from the Airin district of Osaka City. RESULTS: The patients with HCC had a history of long stay (mean: 25 years) in the district and many infectious opportunities and most of them were malnourished. The main causes of liver disease in the patients with HCC were hepatitis C virus (HCV) (77%), alcohol abuse (73%), and the combination of HCV and alcohol abuse (50%). Serum HCV RNA concentration was 10(5.8 +/- 0.9) copies/50 microliters in the 21 HCC patients and 10(6.5 +/- 0.7) copies/50 microliters in the 14 LC patients (p < 0.02). Six HCC patients (20%) were positive for the GB virus C/Hepatitis G virus (GBV-C/HGV) RNA in association with HCV or hepatitis B virus (HBV). Only 2 patients with HCC underwent the curative operations and most of the HCC cases were in progressed stages. CONCLUSIONS: A long stay in a hygiene-poor environment increases the opportunity for infection in homeless people. The causative agents in the HCC and LC patients were mostly HCV, alcohol abuse, and a combination of the two. Since the quantification of HCV-RNA in the HCC patients was lower, the high level of HCV-RNA may not be a risk factor for the development of HCC. GBV C/HGV may not also. The reversion to former healthy living conditions and reduction in alcohol consumption as soon as possible may contribute to low incidence of HCC and save the tax dollar expenditures among homeless people. PMID- 10370676 TI - The role of colchicine in prevention of hepatic cirrhosis induced by carbon tetrachloride. AB - BACKGROUND/AIMS: The present study was undertaken to determine whether colchicine has a beneficial effect in the prevention of hepatic cirrhosis when it is given simultaneously with CCl4. METHODOLOGY: Wistar rats were employed as experimental animals and divided into 6 groups: Group I received saline solution, Group II, saline solution and mineral oil; Group III, colchicine (10 micrograms/100 g) and mineral oil; Group IV, colchicine (10 micrograms/100 g) and CCl4; Group V, colchicine (5 micrograms/100 g) and CCl4; and, Group VI received saline solution and CCl4. The effect of colchicine was evaluated by liver function tests, serum total proteins, electrolytes and histological evaluation. RESULTS: The results demonstrated higher values of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and total bilirubin in groups IV and V when compared with group VI (p < 0.05). No difference between group VI and groups IV and V was observed in histological evaluation, serum total proteins and electrolytes (p < 0.05). CONCLUSIONS: Colchicine, as given in this study, did not have any protective effect in the prevention of cirrhosis induced by carbon tetrachloride. PMID- 10370677 TI - Intrahepatic interleukin-2 with chemotherapy for unresectable liver metastases: a randomized multicenter trial. AB - BACKGROUND/AIMS: A pilot study of Interleukin-2 (IL-2) with chemotherapy for unresectable colorectal liver metastases revealed a favorable response rate (76%). This prospective, randomized, multicenter study was conducted to evaluate the efficacy of this treatment protocol. METHODOLOGY: Over a period of 32 months, 46 patients with unresectable liver metastases were randomly assigned to 1 of 3 treatment groups: group A: chemotherapy alone, group B: chemotherapy plus high dose, intermittent IL-2 (2.1 x 10(6) U twice weekly) or group C: chemotherapy plus low-dose, continuous IL-2 (7 x 10(5) U daily). Treatment continued for 4 weeks in the hospital and on an outpatient basis according to the clinical response. No crossover between treatment arms was permitted. RESULTS: IL-2 combined with chemotherapy produced a higher complete and partial response rate of 40% in group A, 60% in group B, and 78% in group C. Toxicity related to IL-2 included fever, chills, malaise, and eosinophilia. CONCLUSIONS: Hepatic arterial infusion of chemotherapy plus IL-2 resulted in an increased tumor response when compared with chemotherapy alone. To confirm the efficacy of this treatment protocol, we have started a large-scale, randomized, multi-institution trial. PMID- 10370678 TI - Intra-arterial versus systemic chemotherapy for non-operable hepatocellular carcinoma. AB - BACKGROUND/AIMS: To compare intra-arterial (regional) hepatic chemotherapy with doxorubicin, to the systemic (intravenous) one in patients with non-resectable (Stage IVA) hepatocellular carcinoma. METHODOLOGY: Seventy-two patients with inoperable hepatocellular carcinoma were randomized to receive doxorubicin 50 mg/m2 as a bolus infusion either via an implantable intra-arterial catheter (Group A) or as systemic chemotherapy (Group B) every 21-28 days. RESULTS: Patients of Group A had a higher rate of objective and subjective remissions and Karnofsky performance status improvement in comparison to Group B. The mean survival was 7 months (range: 2-16) for Group A and 6.5 months (range: 1-13) for Group B, but this difference was not statistically significant. The quality of life remained at an acceptable level until death in both groups. CONCLUSIONS: A slight but not statistically significant superiority of intra-arterial chemotherapy against the systemic one is concluded. PMID- 10370679 TI - Prevention of infectious complications after transjugular intrahepatic portosystemic shunt in cirrhotic patients with a single dose of ceftriaxone. AB - BACKGROUND/AIMS: Patients with cirrhosis of the liver are prone to bacterial infections. Therapeutic interventions such as endoscopic sclerotherapy increase the risk of bacterial infections in these patients. Following insertion of a transjugular intrahepatic portosystemic shunt (TIPS), the incidence of severe bacterial infections was recently shown to be 20% after elective procedures. This finding suggests antibiotic prophylaxis with the TIPS procedure. Antibiotic prophylaxis using cefotiam or cefotaxime/ampicillin did not significantly reduce infectious complications. The aim of the present study was therefore to investigate the efficacy of two different doses of a long-acting cephalosporin in prevention of bacterial infection after TIPS. METHODOLOGY: Eighty-two patients with cirrhosis (age: 52 +/- 2 years) who underwent elective TIPS were randomized to receive a single i.v. dose of either 1 g or 2 g Ceftriaxone 1 hour before the intervention. Patients with evidence of or suspected infections and patients on antibiotic therapy within 7 days prior to TIPS were excluded. Body temperature was monitored t.i.d. for 1 week and white blood count (WBC) and C-reactive protein (CRP) were determined before TIPS and 1 day and 1 week after TIPS. RESULTS: Only 2 of 82 patients (2.6%) showed signs of infection following TIPS insertion: One of 40 patients receiving 1 g Ceftriaxone and 1 of 42 patients receiving 2 g Ceftriaxone prior to TIPS developed temperature > 38.5 degrees C. In the latter patient this was due to pneumonia. This patient received antibiotic treatment with imipenem for 10 days. Temperature in the other patient normalized within 12 hours and he did not require antibiotic treatment. No significant differences in temperature, WBC and CRP between the different doses of Ceftriaxone were observed. CONCLUSIONS: Prophylactic treatment with Ceftriaxone reduces the reported incidence of bacterial infections after TIPS in patients with cirrhosis of the liver. Prophylaxis with 1 g Ceftriaxone seems as efficacious as 2 g. PMID- 10370680 TI - Initial daily interferon administration can gain more eradication of HCV-RNA in patients with chronic hepatitis C, especially with serum intermediate viral load. AB - BACKGROUND/AIMS: We studied the effect of initial daily administration of interferon for the treatment of chronic hepatitis C, especially in patients with intermediate viral load. METHODOLOGY: Consecutive patients who met the inclusion criteria were randomly enrolled into two groups in this study. All patients analyzed could be treated with interferon-alpha for 6 months. Patients in group A were administered 6 million units of interferon-alpha subcutaneously daily initially for 2 weeks and then thrice weekly. Patients in group B were treated with the same dose of interferon-alpha thrice weekly from the first administration. We decided the criteria of complete remission as the absence of serum HCV-RNA at both points of the end of interferon treatment and 6 months later. RESULTS: Due to the relationship between the efficacy and serum viral load, we decided the criteria of the intermediate load as the quantitative value of serum HCV-RNA to be not lower than 10(5.0) and not higher than 10(6.5) copies/ml. Seventy-six and 78 patients, whose genotype and quantitative value of serum HCV-RNA could be measured before treatment, were analyzed in group A and B, respectively. The rate of complete remission in group A (40.8%) was higher than that in group B (25.6%), significantly (p = 0.046). In the intermediate viral load group, the rate of complete remission in group A (52.3%) was significantly higher than that in group B (29.3%) (p = 0.045). In the patients with genotype 1 b virus, the rate of complete remission had a tendency to be higher in group A (33.3%) than in group B (17.4%) (not significant). In the patients with genotype 2, the rate of complete remission was higher in group A (77.8%) than in group B (41.2%) (significant, p = 0.041). CONCLUSIONS: These results suggest that the initial daily interferon administration is necessary to gain a higher rate of serum HCV-RNA eradication in patients with intermediate viral load in chronic hepatitis C. PMID- 10370681 TI - An approach for difficult hepatectomy--retrograde hepatectomy in 29 patients with liver malignant tumor. AB - BACKGROUND/AIMS: Resection remained the best treatment for malignant liver tumor. However, it is difficult to resect a tumor which is huge and tightly invaded or adhered to the surrounding organs by classical procedures because of poor exposure. The purpose of the present study was to verify that retrograde hepatectomy was an acceptable approach. METHODOLOGY: Retrograde hepatectomy means that the operative procedure is reversed as compared with classical methods. Transection of the liver parenchymal was performed first, isolating adhesions between the resected liver and diaphragm or partial phrenectomy followed, and then after cutting corresponding ligaments, the liver tumor was removed. If the adjacent organs were invaded or adhered too tightly to be separated, they were removed with the resected liver. This approach was adopted in 29 patients with liver malignancy (group A) for difficult hepatectomy from June 1994 to June 1997. In the same period, classical hepatectomy was performed in 13 patients used as a control group (group B). The differences between these two groups were analyzed. RESULTS: When group A was compared with group B, the operative mortality was 0% versus 7.7% (p > 0.05), the operative time was shorter, being 175.9 +/- 49.7 min (range: 150-250 min) versus 251.9 +/- 66.9 min (range: 180-360 min) (p < 0.05), the estimated intra-operative blood loss being 1430.0 +/- 807.6 ml (600-4200 ml) versus 2907.7 +/- 1497.9 ml (800-7000 ml) (p < 0.05), and the incidence of post operative complications was lower (p < 0.05). CONCLUSIONS: Retrograde hepatectomy is an alternative method to classical hepatectomy and suitable for resection of localized huge liver tumor when the exposure is inadequate by classical approach, particularly when the tumor adheres or invades closely to the diaphragm and/or the surrounding structures. PMID- 10370682 TI - Emergency liver resection for ruptured hepatocellular carcinoma complicating cirrhosis. AB - BACKGROUND/AIMS: From a consecutive series of 51 patients surgically treated from January 1993 to August 1997 for hepatocellular carcinoma (HCC) complicating cirrhosis, 6 subjects (12%) presented with acute hemoperitoneum due to spontaneous rupture of the tumor: 3 patients were suffering from chronic hepatitis C, 2 were affected by alcoholic cirrhosis, and one by chronic hepatitis B. The present paper reports experience of the treatment of ruptured HCC complicating cirrhosis in 6 patients undergoing emergency hepatectomy. METHODOLOGY: Hemoperitoneum was successfully diagnosed pre-operatively with the combination of abdominal ultrasound (US) and paracentesis. All subjects had a known history of chronic liver disease, but undiagnosed HCC. Child-Pugh classification assessed the hepatic functional reserve to predict operative risk. Surgical indication was based on hemodynamic instability and/or persistent bleeding. Time from admission to operation was recorded as well as tumor site, size and number, the site of bleeding, and the duration of surgery and hepatic devascularization. Tumor location was defined according to segmental anatomy. All patients underwent one-stage liver resection (segmentectomy VII-VIII in one patient; non-anatomical wedge resections in 5). Operative mortality was defined as death within 30 days of surgery. RESULTS: No intra-operative death occurred. In 4 patients the post-operative course was uneventful. Two patients died 2 weeks after surgery from liver failure (one patient) eventually complicated by renal failure (one patient). Three patients are alive and 2 of them disease-free at 24 months after surgery, whilst one patient has died from liver failure 21 months after surgery in the presence of intrahepatic recurrence of HCC. CONCLUSIONS: Present experience, combined with a literature review on 755 ruptured HCC cases, indicates that emergency liver resection is feasible in patients with limited tumor and preserved liver function (Child-Pugh A or B grade); surgical resection is the only procedure possibly associated with long-term survival, as shown by 4/6 patients of ours surviving more than 12 months, with 2 subjects disease-free at 24 months. Conservative management, such as surgical/radiological devascularization, packing or plication, can be conducted on high risk patients, though long-term survivors have not been reported. PMID- 10370683 TI - Extended hepatectomy with ePTFE graft vena caval replacement and hepatic vein reconstruction: a case report. AB - A 69 year-old man with a history of thoracoplastic surgery for pulmonary tuberculosis, who required a blood transfusion and subsequently tested positive for hepatitis C virus, developed a right hypochondrial mass, swelling of the lower extremities and malaise. A huge hepatocellular carcinoma invading the suprahepatic vena cava with tumor thrombi was diagnosed radiographically. An extended right hepatectomy with supra- to retrohepatic IVC resection was performed in an en bloc fashion using a centrifugal pump for hepatic vascular exclusion (HVE). The supra- to retrohepatic IVC was replaced with an expanded polytetrafluoroethylene (ePTFE) graft, 20 mm x 10 cm in size, and the left hepatic venous confluence was reconstructed. Twenty-one months after surgery, the patient is in good condition without recurrence of tumor. PMID- 10370684 TI - The non-surgical treatments of hepatocellular carcinomas in a patient with severe hemophilia A. AB - We recently treated a patient with multiple hepatocellular carcinomas (HCC) who had severe hemophilia A and thrombocytopenia secondary to cirrhosis. He was not a candidate for surgery because of his liver failure. Transcatheter arterial chemoembolization and percutaneous ethanol injection therapy were successfully performed without complications by maintaining factor VIII levels above 40%. We believe that the non-surgical treatment of HCC can be performed safely in patients with coagulation disorders, by the appropriate administration of coagulation factors, despite thrombocytopenia and/or elongation of the prothrombin time from cirrhosis. PMID- 10370685 TI - Surgical management for lymph node recurrence of resected fibrolamellar hepatocellular carcinoma: a case report. AB - Fibrolamellar hepatocellular carcinoma (FLHCC), which is quite uncommon in Japan, is known to be frequently associated with lymph node metastasis in Western countries. Herein, we describe a case of a 25 year-old Japanese woman with recurrent FLHCC in the lymph nodes after undergoing right hepatic lobectomy. She underwent a second operation for removal of a recurrent celiac lymph node tumor 23 months after the initial operation. In Japan, the frequency of lymph node metastasis in ordinary hepatocellular carcinoma is only 1.6%, whereas 3 out of 9 (33%) reported domestic FLHCCs including this case had lymph node metastasis. The surgical management of lymph node metastasis in FLHCC is discussed. PMID- 10370686 TI - An approach to the rational use of steatotic donor livers in liver transplantation. AB - Steatosis or fatty change is a common finding in donor liver biopsies during liver transplantation, and seems to be more frequent than in the general population. Fat can be stored in hepatocytes within macrovacuoles (macrosteatosis) or microvacuoles (microsteatosis), with different degrees of severity. Higher degrees of both macro and microsteatosis may increase the severity of the ischemia-reperfusion lesion producing an initial poor function in the recipient. Different pathogenic mechanisms have been investigated. However, only severe macrosteatotic (> 60%) grafts have been associated with primary non function, and are universally rejected for transplantation. While donor livers with any severity of microsteatosis do not influence recipient survival and can be safely implanted, donor livers with moderate to severe macrosteatosis (30-60%) have a relative risk of primary non-function and should be considered for transplantation in the absence of other known risk factors. A protocol with a rational use of these steatotic livers is suggested. PMID- 10370687 TI - Laparoscopic pancreatic surgery: indications, techniques and preliminary results. AB - BACKGROUND/AIMS: The purpose of this study was to evaluate feasibility, effectiveness and reproducibility of laparoscopy in the diagnosis and management of pancreatic diseases. METHODOLOGY: At the Second Department of Surgery of the "Regina Elena" Institute for Cancer Research, between June 1995 and June 1997, possibilities of the laparoscopy in diagnosis and staging of pancreatic malignancies were studied. Twenty-four consecutive cases of pancreatic adenocarcinoma were observed: 10 patients were pre-operatively considered unresectable and 14 cases were submitted to a laparoscopic study. Moreover, in the period from January 1996 to June 1997, a study on the therapeutical potentialities of laparoscopy was performed, attempting 5 pure or assisted laparoscopic pancreatic resections. RESULTS: Three cases (21.4%) out of the 14 evaluated were found to be metastatic (liver and peritoneum); 11 patients were successfully resected with an open standard procedure. Laparoscopy allowed a correct staging in all cases, even in the 21.4% of ultrasonography (US), TC and angiographic false negatives. The 5 operative procedures consisted of one resection of a cystadenoma, one resection of a head cyst, one Wirsung lithotomy with pancreaticojejunostomy for lithiasic chronic pancreatitis, and two body-tail splenopancreatectomies (1 cystadenoma and 1 adenocarcinoma). Satisfactory early results were obtained in all these cases. CONCLUSIONS: Laparoscopic diagnosis and staging of pancreatic cancer is a valuable method which allows a very high accuracy. Pancreatic resections, even if feasible, are still to be considered an experimental procedure. PMID- 10370688 TI - Pathophysiology after pylorus-preserving pancreatoduodenectomy: a comparative study of pancreatogastrostomy and pancreatojejunostomy. AB - BACKGROUND/AIMS: Pylorus-preserving pancreatoduodenectomy using the pancreatogastrostomy technique may result in pancreatic exocrine insufficiency and obstruction of the pancreatic duct. A prospective randomized comparison of pancreatogastrostomy and pancreatojejunostomy was therefore performed to assess pathophysiologic changes after pylorus-preserving pancreatoduodenectomy. METHODOLOGY: The study population consisted of 23 patients (pancreatogastrostomy: 10, pancreatojejunostomy: 13) who were observed for 2 years. RESULTS: Neither physical condition (dietary intake, body weight, performance status, and frequency of bowel movements) nor nutritional parameters (serum levels of total protein, albumin, total cholesterol, and cholinesterase) differed significantly between the two groups; these parameters recovered to pre-operative levels within 1 year in both groups. Changes in pancreatic function diagnosis (PFD) test results were similar between the two groups. The glucose tolerance test results revealed deterioration of glucose tolerance in 2 patients (20%) in the pancreatogastrostomy group and 3 patients (23%) in the pancreatojejunostomy group. In 2 of 3 patients in each group with non-dilated pancreatic ducts before surgery, the pancreatic ducts dilated after surgery. Diabetes developed after surgery in one such patient in each group. No significant differences were observed between the two groups with respect to changes in glucose tolerance test results and the diameter of the pancreatic duct. CONCLUSIONS: This prospective randomized study demonstrates no difference in pathophysiologic changes between patients undergoing pancreatogastrostomy or pancreatojejunostomy after pylorus preserving pancreatoduodenectomy, at least in the first 2 years. PMID- 10370689 TI - Ampullary carcinoma: prognostic significance of ploidy, cell-cycle analysis and proliferating cell nuclear antigen (PCNA). AB - BACKGROUND/AIMS: The aim of the present study is to assess the nuclear DNA ploidy patterns, the fraction of cells in the various phases of the cell cycle as determined by flow cytometry and to evaluate Proliferative cell-nuclear antigen (PCNA) expression in order to examine the relationships between phase-two molecular factors, clinicopathological aspects and outcome of patients with cancers of the ampulla of Vater. METHODOLOGY: Paraffin-embedded specimens from 18 cases of cancers of ampulla of Vater radically resected between 1985 and 1995 were analyzed by flow-cytometry and immunohistochemical staining with monoclonal antibody to the PCNA. The relationships between cell-proliferation kinetics, PCNA positive cancer cells, clinicopathological findings and the clinical course were evaluated. RESULTS: Pathologist reports documented 17 papillary adenocarcinomas and one case of mucinous carcinoma. According to the TNM classification, 4 patients were in stage I, 7 in stage II and 7 in stage III. Locally advanced ampullary tumors (T3-T4) had a significantly worse prognosis (p = 0.01); survival at 3 and 5 years for stage I-II patients (11 cases) was 90% and 79% as compared to 42% and 42% for patients with stage III (8 cases), respectively (p = n.s.). Thirteen cancers (72%) were diploid and 5 (28%) aneuploid. Patients with aneuploid tumors were younger (mean age: 59 years) than patients with diploid tumors (mean age: 66 years; p = 0.04). No significant correlation was found between size of the tumor (T), lymphnodal status (N), grading (G) or aneuploidy. Difference in terms of survival between aneuploid and diploid patients was relevant (16 vs. 121 months) but, due to the small number of cases, was not statistically significant (p = n.s). The mean value of S-phase fraction (SPF) was 14.8%. PCNA positive rate significantly correlates with size of the tumor (T1-T2 vs. T3-T4; p = 0.03). Actuarial overall survival resulted in 70%, 63% and 31% at 1, 5 and 10 years, respectively. The high rate of diploidy (72%) supports the relative benign behavior of ampullary cancers. CONCLUSIONS: PCNA positive rate significantly correlates with size of the disease. Aneuploidy, although without significant prognostic value, correlates well with survival. Because of the wide range of all variables, more data are needed to establish the relationships between pathological factors, DNA ploidy and PCNA rate and their significance as molecular predictors of prognosis in ampulla of Vater cancers. PMID- 10370691 TI - Pancreatic metastasis from renal cell carcinoma causing massive gastrointestinal bleeding in von Hippel-Lindau disease. AB - Patients with von Hippel-Lindau disease (VHLD) may develop pancreatic lesions, including cysts, serous cystadenomas and islet cell tumors. However, only a few cases of pancreatic metastasis have been reported. We present a case of VHLD with multiple pancreatic metastases from renal cell carcinoma. One of the metastases invaded the duodenum, causing massive bleeding. PMID- 10370690 TI - Three generations of hereditary chronic pancreatitis. AB - The patient was a 22 year-old male. Hereditary chronic pancreatitis was suspected as a diagnosis since his mother's uncle had been operated on for chronic pancreatitis 14 years previously at the age of 64 years and his mother had been operated on for chronic pancreatitis with calculi 5 years previously at the age of 40 years. Surgery was needed, since: 1) he had experienced abdominal pain for 8 years; 2) endoscopic retrograde cholangiopancreatography (ERCP) revealed a marked irregular dilatation in the main pancreatic duct and a marked irregular dilatation and protein plugs in the ductule of the tail of the pancreas; and, 3) pancreatic functional diagnostic (PFD) test examination showed a 75% decrease in exocrine function. If a surgical procedure had not been performed, the patient would likely have experienced calculi formation in the pancreas and a further decrease in exocrine function. Since the patient was very young and had many protein plugs in the dilated ductule of the tail of the pancreas, we decided to perform a spleen-preserving Puestow's procedure with removal of the tail of the pancreas. Clinical and pathological findings of hereditary pancreatitis are reviewed. PMID- 10370692 TI - Rosai-Dorfman disease (sinus histiocytosis with massive lymphadenopathy) of the pancreas: first case report. AB - Rosai-Dorfman Disease is a histiocytic proliferative disorder which primarily affects lymph nodes. Extranodal involvement occurs in about one half of the patients and the head and neck area represents the region most commonly involved. We present the case of a 48 year-old female who was found with a pancreatic mass during evaluation for abdominal pain. She underwent a distal pancreatectomy and splenectomy. Her pathology showed sinus histiocytosis with massive lymphadenopathy (SHML) involving the pancreas and lymph nodes and focally the spleen. The histiocytes characteristically contained one or more viable lymphocytes in the cytoplasm. The lymphocytes had penetrated the cytoplasm in a process known as "emperipolesis", where the lymphocytes continued to have free movement in the histiocyte. The histiocytic cells were positive with S-100 protein and CD68. Rosai-Dorfman Disease (SHML) can affect the peripancreatic lymph nodes with possible secondary pancreatic involvement and present as a pancreatic mass. PMID- 10370693 TI - Economic evaluations of gastric and pancreatic cancer. AB - The total cost of cancer care in the US is about $146 billion, of which pancreatic cancer comprises $2.6 billion (1.8% of the total) and gastric cancer comprises $1.8 billion (1.3%). We have reviewed published studies presenting economic analysis of treatment or follow-up for patients with pancreatic or gastric cancer. Relatively few studies report on economic evaluations of pancreatic cancer care. There are also few economic studies for gastric cancer, although we identified three cost-effectiveness analyses. In general, economic analyses in these areas are relatively unsophisticated, relying on charge data or simple multipliers (e.g., average cost per day in the hospital multiplied by days in the hospital), and are often limited to in-hospital costs (particularly studies for pancreatic cancer). A wide range of costs is included in these studies and a variety of methodologies for assigning costs are used, making comparisons between studies difficult. Future health economics research in this area should evaluate the costs and effectiveness of alternative practice patterns for gastric and pancreatic cancer; conduct additional cost-effectiveness analyses of chemotherapeutic interventions; consider quality of life, survival, stage at diagnosis, patient-borne costs, and complications of therapy; and, take advantage of administrative data from large populations. PMID- 10370695 TI - Xanthoma of the stomach--some morphometrical peculiarities and scanning electron microscopy. AB - BACKGROUND/AIMS: "Lipid Iceland's" in gastric mucosa, a rare finding in gastroenterology, is described in this paper. The aim of our investigation was to evaluate the frequency of the so-called "gastric xanthoma" in biopsy specimens and to describe the picture elicited by scanning electron microscopy. METHODOLOGY: We investigated cell characteristics histochemically, morphometrically and with scanning electron microscopy. RESULTS: The frequency of xanthoma in our gastrobiopsy material was 0.018% (4 of 21,650 cases) over a 6 year period. The picture of stomach xanthoma given by scanning electron microscopy is very typical of the condition and is called "rolling stones". CONCLUSIONS: We suggest that this diagnostic method warrants further investigation. PMID- 10370694 TI - Lymphoepithelioma-like carcinoma of the stomach: a subset of gastric carcinoma with distinct clinicopathological features and high prevalence of Epstein-Barr virus infection. AB - BACKGROUND/AIMS: To evaluate the clinicopathological features of lymphoepithelioma-like carcinoma of stomach in Taiwan. METHODOLOGY: Of 379 patients with gastric adenocarcinoma, from 1993 to 1996, 6 of them with lymphoepithelioma-like carcinoma of stomach were retrospectively studied. RESULTS: Five patients were females and one patient was male. Their age ranged from 51-75 years with a mean age of 61.5 years. Endoscopically, 2 patients were initially diagnosed as early gastric cancer and the other 4 were diagnosed as advanced gastric cancer. Three patients had tumors located in the lower third of the stomach, while the other three tumors were located in the middle and upper third. Two tumors invaded into the serosal layer and the other four lesions were confined at submucosal and muscular layers. Using the in situ hybridization method, all 6 patients (100%) had positive nuclear Epstein-Barr virus-encoded small RNA signals in the tumor cells but not in the surrounding lymphoid stroma and non-neoplastic gastric mucosa. Helicobacter pylori was found in 4 (66.7%) of the cases. The mean follow-up period of the 6 patients was 27 months. Five patients were free of the disease. Lymph node involvement and mesenteric implantation was noted in one patient in which cancer recurred 1 year after gastrectomy. CONCLUSIONS: Lymphoepithelioma-like carcinoma of stomach in this study revealed a female predominance, preferential localization in the proximal part of the stomach, better prognosis, and a high association with Epstein-Barr virus infection. PMID- 10370696 TI - Survival in early gastric cancer: multivariate analysis on 72 consecutive cases. AB - BACKGROUND/AIMS: Few reports from the Western hemisphere have investigated the impact of pathological features and surgical modalities on the prognosis of patients affected by early gastric cancer (EGC). In particular, the extent of lymphadenectomy (limited vs. extended) and the type of gastric resection (subtotal vs. total) remain controversial issues in the management of EGC. The aim of this study was to identify factors influencing prognosis in patients affected by EGC. METHODOLOGY: Hospital records and pathological specimens of 72 patients with EGC undergoing resective surgery during the period 1981-1995 were retrospectively reviewed. Patient status was determined by follow-up examination or by telephone contact. Univariate and multivariate analysis was used to calculate the 5-year survival probabilities with respect to the following variables: age (< or = 65, > 65), sex, depth of invasion (mucosal, submucosal) tumor location (upper, middle and lower third), gross appearance (type I, type II and type III), size (< or = 1.5 cm, > 1.5 cm), presence or absence of lymph node metastasis, histological type (intestinal, diffuse), extent of lymphadenectomy (limited or extended), and type of gastrectomy (total or distal subtotal). Survival was the outcome variable studied. RESULTS: Multivariate logistic regression analysis showed that age, nodal involvement and depth of invasion were independently associated with poor survival. CONCLUSIONS: Results showed a significant dominance of host- and tumor-related factors over the type of surgical procedure on prognosis of EGC patients. PMID- 10370697 TI - Segmental gastrectomy for early cancer in the mid-stomach. AB - BACKGROUND/AIMS: We modified the surgical procedure for segmental gastrectomy, which is normally used for peptic ulcers, to treat early gastric cancer of the mid-stomach. In this paper, we describe the surgical technique and its results. METHODOLOGY: The location of the tumor was confirmed by intra-operative endoscopic examination. An area 2 cm proximal and distal to the tumor was marked with sutures. Firstly, the lymph nodes were dissected from around the perigastric and along the left gastric and common hepatic arteries. Then, a segmental gastrectomy was performed. The greater omentum, omental sac, and vagal nerve, including the hepatic, pyloric and celiac rami, were left intact. An end-to-end gastrogastrostomy was performed using Gambee's sutures and 4-0 monofilament polydioxanone. Gastric drainage was not necessary. RESULTS: We performed segmental gastrectomies on 30 patients. Tumors less than 1 cm in diameter were found in 4 patients; 1.1-2 cm in 14, 2.1-5 cm in 11, and a tumor exceeding 5.1 cm in one patient. The cancer was confined to the mucosa in 23 patients; in the other 7, it had penetrated the submucosa. No lymph node metastases were found but 2 patients had microscopic invasion or permeation of the lymphatic vessels. One patient required post-operative balloon dilation of the pyloric sphincter for delayed gastric emptying. The remaining patients had no post-operative complications. To date, 29 patients, excluding one who died in a traffic accident, have survived disease-free for a mean of 30 months (range: 7-51). Their body weight and dietary volume returned to pre-operative levels within 12 months of surgery. CONCLUSIONS: Patients who underwent segmental gastrectomy have had a reasonably good quality of life in the post-operative follow-up to date. PMID- 10370698 TI - Relationship between enterogastric reflux estimated by scintigraphy and the presence of Helicobacter pylori. AB - The aim of the study is assessment of the relationship between enterogastric reflux and the presence of Helicobacter pylori infection as factors that cause gastritis, peptic ulcer and adenocarcinoma ventriculi. The study was performed in 52 patients with different digestive disorders, using gamma camera, during 90 min (1 frame/min) after intravenous injection of 185 MBq 99mTc-dietil IDA in the cubital vein. According to time/activity curves from the region of hepatobiliary system and stomach, index of enterogastric reflux (EGR) was assessed. There was no correlation between the presence of Helicobacter pylori and EGR (r = 0.181, df = 52, p > 0.05). However, Helicobacter pylori was present more frequently in the patients with positive EGR (p < 0.01), but there were no significant differences (p > 0.05) in reflux value in patients with either positive or negative findings of Helicobacter pylori. PMID- 10370699 TI - Post-operative chemotherapy in non-curative gastrectomy for advanced gastric cancer. AB - BACKGROUND/AIMS: The definitive effects of post-operative chemotherapy for prolonging survival in patients with non-curative gastrectomy for advanced gastric cancer have not been established. METHODOLOGY: Eighty-three patients with advanced gastric cancer who underwent non-curative gastrectomy were divided into 49 patients with post-operative chemotherapy (chemotherapy group) and 34 patients without post-operative chemotherapy (control group). Chemotherapy regimens were as follows: oral 5-fluorouracil (5-FU) alone (n = 22), intravenous mitomycin (MMC) plus 5-FU (n = 20), intravenous methotrexate (MTX) plus 5-FU (n = 3), intravenous cisplatin plus 5-FU (n = 2), and hepatic arterial infusion of 5-FU plus oral 5-FU (n = 2). No prior chemotherapy or radiation therapy was given. RESULTS: Although the age in the control group (mean: 71.9 years) was significantly older than in the chemotherapy group (mean: 66.1 years), there were no significant differences in the other clinical and pathological background data between the two groups. The 1-year survival rate in the chemotherapy group (71.4%) was significantly higher than in the control group (50.0%). However, the 3-year and 5-year survival rates did not significantly differ in the chemotherapy group versus the control group, 30.6% vs. 32.4% and 24.5% vs. 32.4%, respectively. Although a significant difference did not exist between the two groups, median survival after operation in the chemotherapy group (20.5 months) was longer than that in the control group (16.2 months). Furthermore, median survival of patients with peritoneal dissemination in the chemotherapy group (16.4 months) was significantly longer than that in the control group (7.7 months). CONCLUSIONS: Post-operative chemotherapy may contribute to prolonged survival in patients with non-curable advanced gastric cancer, even when patients had peritoneal dissemination. However, the long-term survival rate was not improved by post-operative chemotherapy. More aggressive chemotherapy may be needed to improve the long-term prognosis for such patients. PMID- 10370700 TI - Transcutaneous electrogastrography: a non-invasive method to evaluate post operative gastric disorders? AB - BACKGROUND/AIMS: With the development of high-performance computer programs, transcutaneous electrogastrography has experienced a renaissance in the last few years and is widely recommended as a non-invasive diagnostic tool to evaluate functional gastric disorders. We assessed the clinical value of electrogastrography in symptomatic and asymptomatic patients after a variety of procedures of the upper gastrointestinal (GI) tract. METHODOLOGY: Electrogastrography tracings were recorded with a commercially available data logger using a recording frequency of 4 Hz. A standard meal was given between a 60 min preprandial and a 60 min postprandial period. The following parameters were analyzed pre- and postprandially utilizing Fourier and spectral analysis: Regular gastric activity (2-4 cycles/minute), bradygastria (0.5-2 cycles/minute), tachygastria (4-9 cycles/minute), dominant frequency and power of the dominant frequency. Nineteen asymptomatic healthy volunteers served as a control group. Forty-nine patients, who had undergone upper intestinal surgery, were included in the study (cholecystectomy n = 10, Nissen fundoplication n = 10, subtotal gastrectomy n = 8, truncal vagotomy, and gastric pull-up as esophageal replacement n = 6). Twenty of these patients complained of epigastric symptoms post-operatively, while 12 of these 20 patients also had a scintigraphic gastric emptying study with Tc99m labeled semisolid meal. RESULTS: Preprandial gastric electric activity was between 2 and 4 cycles/minute in 60-90% of the study time in healthy volunteers. In all study groups the prevalence and power of normal electric activity increased significantly after the test meal (p < 0.001). After cholecystectomy, Nissen fundoplication, subtotal gastrectomy or vagotomy and gastric pull-up pre- and postprandial gastric electric activity showed a greater variability compared to normal volunteers (p < 0.05), but no typical electrogastrography pattern could be identified for the different surgical procedures. There was no significant difference in the electrogastrography pattern between asymptomatic and symptomatic patients and patients with normal or abnormal scintigraphic gastric emptying curves. CONCLUSIONS: There is no specific electrogastrography pattern to differentiate between typical surgical procedures or epigastric symptoms. To date, electrogastrography does not contribute to the diagnosis and analysis of gastric motility disorders after upper intestinal surgery. PMID- 10370701 TI - Control of solitary gastric fundal varices and portosystemic encephalopathy accompanying liver cirrhosis by balloon-occluded retrograde transvenous obliteration (B-RTO): a case report. AB - In a patient with liver cirrhosis complicated by solitary gastric fundal varices and portosystemic encephalopathy, Balloon-occluded retrograde transvenous obliteration (B-RTO) of the varices was performed. The gastric varices were decreased in size 2 weeks after treatment and had not recurred after 1 year. B RTO successfully occluded the portosystemic shunt (gastrorenal shunt). Accordingly, the patient's blood ammonia levels, total bile acid level, and 15 min retention rate of indocyanine green decreased, and his hepatic encephalopathy improved. However, since consecutive increase in blood flow through the portal collateral vessels except for gastrorenal shunt vessel at 6 months and 1 year after B-RTO was noted, further careful follow-up may be required. PMID- 10370702 TI - Ten-year survival after pancreatoduodenectomy for advanced gastric cancer--report of two cases. AB - We performed pancreatoduodenectomy for 5 patients with gastric cancer, and here we present 2 who have survived for more than 10 years. Patient one had a large antral tumor tightly adherent to the head of the pancreas. Pancreatoduodenectomy with lymph node dissection was performed. Pathologic examination of the resected specimen revealed that the tumor was a well differentiated adenocarcinoma invading the duodenum, but not the pancreas. Patient two had an infrapyloric lymph node metastasis invading not only the pancreatic head, but also the duodenocolic ligament and the transverse mesocolon. Pancreatoduodenectomy and right hemicolectomy with lymph node dissection were performed. Pathological examination of the resected specimen revealed grade III lymph node metastasis, and invasion of the pancreas by the metastatic infrapyloric lymph node. These results indicate that complete resection of tumor by pancreatoduodenectomy may result in a long survival not only for the patients in whom pancreatic invasion and/or lymph node metastasis is limited, but also for some patients with tumor invading the pancreatic parenchyma and/or of grade III lymph node metastasis. PMID- 10370703 TI - Legal issues in reproductive health care for adolescents. AB - Adolescents need reproductive and sexuality-related health care. State and federal laws affect consent, confidentiality, and payment for services. State minor consent laws allow minors to consent based on the minor's status or the services sought. State and federal laws affect confidentiality of health information generally as well as programs for specific services, such as family planning or drug and alcohol abuse treatment. Medicaid, the new State Children's Health Insurance Program, and categorical funding programs are important funding sources. Adolescents' ability to use these sources for confidential reproductive and sexuality-related services is evolving. However, possible changes in federal law and the transition to managed care in both public and private insurance present significant challenges they may face in the future. PMID- 10370704 TI - Sexuality and sex education at home and school. AB - This article reviews numerous studies of parent-adolescent communication about sexuality and 30 studies of sexuality and HIV education programs. Although parent adolescent communication about sexuality has increased during the last few decades, both the occurrence and the quality of this communication could still be greatly improved. There does not exist a clear simple relationship between greater parent-adolescent communication and less adolescent risk-taking behavior, but both adults and youth believe it is important anyway. Sexuality and HIV education programs do not increase any measure of sexual activity, but some of them with ten common characteristics do reduce sexual risk-taking, either by delaying or reducing sexual behavior or increasing condom use. Thus, these programs can be an effective component in a larger initiative to reduce HIV, other STDs, and unplanned pregnancy in adolescence. PMID- 10370705 TI - An update on contraception for adolescents. AB - Almost one million American adolescents become pregnant each year. Awareness of up-to-date contraceptive methods allows clinicians to appropriately counsel male and female adolescents, preferably prior to the onset of sexual activity. Abstinence, barrier methods, including the female condom and non-latex male condom, and hormonal methods, including oral contraceptive pills, injectables, and subdermal implants, are all suitable for adolescent use. New oral contraceptive pills with lower estrogen content or multiphasic hormone levels may ameliorate side effects, such as headache or breakthrough bleeding. Frank discussion of confidentiality and of the various contraceptive methods and their side effects allows adolescents to choose the best method for them, thus improving the likelihood of adherence. PMID- 10370706 TI - Gynecological health care for adolescents with developmental disabilities. AB - The gynecological health care of adolescents with developmental disabilities presents a unique challenge to care providers. Adolescence is a time of turmoil for many girls and for adolescents with special needs it may seem overwhelming. Patients, their parents and their caregivers need special attention during these changes in their lives. This chapter focuses on how to perform a reproductive health evaluation focusing on the history and physical exam, especially the pelvic exam. Menstrual hygiene and menstrual abnormalities are discussed in detail. Other topics include abuse and abuse prevention, contraception, and cyclical behavior changes. Patience, persistence, and adaptation of the usual examination techniques can lead to a thorough and balanced assessment of the patient and help her, her parents, and her caregivers with the challenges of adolescence. PMID- 10370707 TI - Sexually transmitted diseases: testing and treating. AB - Adolescents remain a group at particular risk for STD acquisition due to a combination of biological and psychosocial factors. Access to care can be an obstacle to seeking appropriate screening and treatment for many adolescents; undetected infection may lead to unwanted sequelae, including pelvic inflammatory disease, chronic abdominal pain, tubal scarring, and increased risk of ectopic pregnancy. With respect to gonorrhea, chlamydia, syphilis, and chancroid, the hope is that improved detection will decrease sequelae by prompting earlier recognition and treatment. In all cases of suspected sexual abuse cultures remain of utmost importance because of the negative consequences associated with a possible false-positive test result. Urine screening in certain settings, such as school-based health centers and juvenile detention centers, remains positive; however, adolescents with a positive test may still require further evaluation to identify HPV and abnormal Pap smear findings, syphilis, and other STDs currently not recognizable with a simple urine screen. PMID- 10370708 TI - Pap smears: screening, interpretation, treatment. AB - Cervical cancer remains an important public health problem in the United States. Cervical dysplasia and rarely carcinoma in situ can occur during adolescence, especially in adolescents who become sexually active with multiple partners or who have been exposed to HPV, especially types 16 and 18. Papanicolaou smear is recommended for any adolescent who has ever been sexually active or exposed to human papillomavirus or is 18 years of age. This article discusses the new classification systems for cytological diagnoses, as well as some of the new types of Pap smear preparations, including the ThinPrep system and computer assisted automated Pap test screening, which may increase the sensitivity of this screening test and reduce the false-negative rate of Pap smears. PMID- 10370709 TI - Menstrual cycle abnormalities: diagnosis and management. AB - In summary, the diagnosis and management of menstrual disorders in adolescent girls is a particular challenge. The clinician must keep an open mind and consider physical conditions as well as psychosocial factors that may play a role in the menstrual dysfunction. Knowledge of congenital and genetic conditions, chronic biomedical as well as psychosomatic disorders, lifestyle choices, and complications of sexual behavior are essential to properly diagnose and treat menstrual disorders in young women. A detailed and confidential medical and psychosocial history and a thorough physical examination, including external genital examination, rectoabdominal or vaginal bimanual examination, and speculum examination (if indicated) are necessary. PMID- 10370710 TI - Amenorrhea in the athlete. AB - Amenorrhea in the athlete is commonly encountered in clinical practice. The work up of an athlete with amenorrhea should include consideration of all the physiological and pathological conditions that give rise to amenorrhea in any adolescent. Delay or failure to recognize and manage these patients may result in the emergence of athletic triad with potential serious consequences of increased stress fractures, scoliosis, and thin body mass. This article reviews amenorrhea in the adolescent athlete with respect to body composition, disordered eating, osteoporosis, psychological factors, warning signs, treatment, and outcomes. Need for further strategies specific to prevention, surveillance, research, health consequences, medical care, and public and professional education is addressed. PMID- 10370711 TI - Pelvic pain. AB - Pelvic pain is a common symptom in the adolescent female. Acute pain may represent a life-threatening situation and torsion, ectopic pregnancy, and PID must be considered. For the young patient who presents with chronic pelvic pain, a multidisciplinary approach is essential to facilitate diagnosis and management. Whenever possible, organic disease such as endometriosis, adhesions, and obstructive malformations should be identified and treated as indicated. Developing a treatment team, recognizing psychosocial and environmental factors, and encouraging long-term relationships are critical components in the care of these patients and in the prevention of recurrent symptom formation and future disability. PMID- 10370712 TI - Vulvar disorders in adolescents. AB - Bacterial and fungal vulvitis, dermatitis, inflammatory dermatoses, secondary drug reactions, viral infections, and a variety of vulvar tumors can all present as primary vulvar problems in adolescents. In addition, systemic disease can present with vulvar involvement. These disorders can be extremely anxiety provoking in adolescent females who are dealing with issues surrounding self image, physical maturation, and sexuality. A detailed history and physical exam can provide many clues to the underlying problem, but sometimes a biopsy is necessary to establish a diagnosis. Consultations from other services, such as dermatology or infectious diseases, can be very helpful. The best approach involves a combination of empathetic reassurance, careful diagnosis, and successful treatment. In order to facilitate optimal care of adolescents with vulvar disorders, a comprehensive review of the literature is presented. PMID- 10370714 TI - Ovarian masses. AB - All types of ovarian tumors seen in the adult population can also be seen in children and adolescents. However, the relative incidence of each entity varies considerably with age. Functional non-neoplastic cysts and masses are much more frequent in the adolescent. Furthermore, in the adolescent age group, neoplasms are more likely to be germ cell rather than epithelial in origin. Fortunately, in children and adolescents only 10% of ovarian tumors are malignant. In diagnostic evaluation of ovarian masses, a complete history and physical examination are of utmost importance. Whenever an ovarian mass, be it physiologic or neoplastic, benign or malignant, is diagnosed in an adolescent female or prepubertal child, every effort must be made to preserve the reproductive function in that female in order to ensure future childbearing. PMID- 10370713 TI - Polycystic ovary syndrome. AB - Many adolescents present with hirsutism and irregular menses. The challenge for the clinician is to distinguish physiologic anovulatory cycles from true menstrual disorders such as PCOS, and to differentiate PCOS from other causes of hyperandrogenism in hirsute adolescents. Common clinical features seen in adolescents with PCOS include hirsutism, acne, menstrual irregularity, and obesity. Biochemical abnormalities include hyperandrogenism, acyclic estrogen production, LH hypersecretion, decreased levels of SHBG, and hyperinsulinemia. Management strategies for a patient with PCOS include treatment of features which may cause distress to the adolescent, such as hirsutism, acne, and irregular menses, and prevention of long-term sequelae. Oral contraceptive pills, antiandrogens, and cosmetic treatments are used to treat hirsutism, acne, and menstrual irregularity. Oral contraceptive pills or medroxyprogesterone acetate are given to prevent endometrial hyperplasia and carcinoma. Counseling about weight loss and nutrition are essential, as weight loss may improve signs of hyperandrogenism and menstrual irregularity and may prevent NIDDM and cardiovascular disease. Insulin-sensitizing agents show promise in terms of decreasing hyperandrogenism, restoring ovulatory cycles, treating infertility, and preventing long-term sequelae. Finally, it is important to recognize that adolescents with PCOS may experience psychological distress because of the clinical manifestations of hyperandrogenism or when confronted with the information that they have a chronic illness. Psychological support should be available for these young women. Future research is likely to further elucidate the pathophysiology of PCOS, identify candidate genes, and clarify which adolescents are at risk for long-term sequelae. Prospective studies are needed to identify which therapies could potentially reduce the risk of infertility, diabetes, cardiovascular disease, and endometrial carcinoma in young women with PCOS. PMID- 10370715 TI - Treatment of diseases of the central nervous system using encapsulated cells. PMID- 10370716 TI - Intracranial endoscopy. AB - Since 1910, when Lespinasse [73] in Chicago was the first surgeon to use an endoscopic device for the treatment of a neurologic disease, various methods of endoscopy have evolved into accepted diagnostic and therapeutic adjuncts of modern neurosurgery. Nevertheless, until recently technical shortcomings of the available endoscopes have prevented the widespread use of neuroendoscopy. However, now, at the end of the 20th century, endoscopes can be regarded as some of the most important instruments for the development of microneurosurgery into the 3rd millennium. The aim of this review of intracranial endoscopy in neurosurgery, which admittedly might not be completely objective in the authors' personal assessment of various endoscopic techniques, is first to depict the historical evolution of neuroendoscopy, second to describe the technical equipment used in intracranial endoscopic neurosurgery, third to characterize the most frequent endoscopic methods in brain surgery, and fourth to indicate how neuroendoscopy might develop in the near future. It will be shown that this ongoing evolutionary process in neuroendoscopy was only possible with the mutual influence of improved diagnostic techniques, increased microanatomical knowledge, refined neurosurgical instrumentation--especially the introduction of the surgical microscope, and endoscopic diagnostic and therapeutic strategies. PMID- 10370718 TI - Recent advances in the treatment of central nervous system germ cell tumors. PMID- 10370717 TI - Chronic deep brain stimulation for movement disorders. PMID- 10370719 TI - Hypothalamic gliomas. PMID- 10370720 TI - Surgical approaches to the anterior fossa, and preservation of olfaction. PMID- 10370721 TI - [Membrane stabilizers (chromones and ketotifen)]. AB - The drugs called "Membrane stabilisers" are composed of cromones (sodium cromoglycate and nedocromil) and Ketotifen. They inhibit the degranulation of mastocytes by a membrane stabilising effect. Ketotifen is distinguished from the cromones by a conjoint antihistamine effect. Nowadays, the indications are for prophylactic treatment of allergic asthma, rhinitis and allergic conjunctivitis and the manifestations of food allergy. Therapeutic efficacy seems to be good in intermittent or mildly persistent asthma, especially in children or young subjects. Secondary effects are rare and generally benign, with reduced contra indications. PMID- 10370722 TI - [Anti-histamines]. AB - The use of anti-histamines remains the therapy of choice for the ensemble of allergic pathologies with a reaginic mechanism. Knowledge of the metabolism of histamine gives an understanding of a number of allergic symptoms and also clarifies the mechanism of action of the anti-histamines. Many molecules are appearing on the market, each with a more precise action on the specific H1 receptors and also carrying other pharmacological specificities of better and better study. Observance of contra-indications, in particular cardio-vascular, allows their use without reserve, whilst knowing that there are always problems of specific tolerance, that are variable from one individual to another, specially on drowsiness. The importance of stopping anti-histamines must be remembered, every time that this is possible, before making an allergic diagnosis by skin tests. PMID- 10370723 TI - [Corticosteroids]. AB - The corticosteroids or glucocorticoids have a preponderant place in the treatment of allergic manifestations. They are used to ward of the inflammatory process triggered and auto-maintained by some mediators (histamine, tryptase, leucotrienes, prostaglandin, ECP, MBP ...) that are released by some cells (mastocytes, basophils, eosinophils ...) during the contest of antigen--antigen receptor site. It is essential to understand the mechanism of action of the glucocorticoids as well as their secondary effects to adapt prescriptions better. PMID- 10370724 TI - [Leukotriene antagonists]. AB - Today, medicinal treatment of asthma over the long term is well codified and involves inhaled corticosteroid therapy, in association or not with beta 2 mimetic substances. Numerous studies have demonstrated the importance of leukotrienes in bronchoconstriction and in the delayed inflammatory reaction of asthma. At the same time, antileukotriene molecules are being introduced. Several are sold in different countries. At present, only one product is available in France: Montelukast. It is officially indicated as a complementary treatment of mild to moderate persistent asthma, not sufficiently balanced by inhaled corticosteroid therapy and as a treatment of exercise-induced asthma. In the future, studies should further define the position of these substances in the long term treatment of asthma. PMID- 10370725 TI - [Adrenaline and beta 2 adrenergics]. AB - Adrenalin is the drug for anaphylactic shock. The product is usable directly by the patient in the form of an auto-injectable kit, by the physician who treats by the sublingual or subcutaneous routes, in a resuscitation unit extra and inter hospital by the intra-veinous route with cardiac monitoring. Precaution have to be observed (in angina especially), but it is without formal contra-indications. The beta 2 adrenergics are non-specific bronchodilators. Two categories are available: short action molecules, crisis drugs, prolonged activity molecules (greater than 12 hours). Several routes of administration are useable: spray or powder (inhalation) sub-cutaneous, taking by mouth. PMID- 10370726 TI - [Therapeutics of the future for allergy]. AB - Future improvement in the therapeutic arsenal in allergy must in part be by improvement of the existing, by the development of new molecules that are more powerful and more selective amongst the antiH1, chromones, Beta-2 mimetics, corticoids as well as the development of new routes of already-know products. Specific Immunotherapy (ITS) will undoubtedly benefit from progress in geni genetics, from improvement in adjuvants and development of new routes of administration. Control of the environment will assist industry for the manufacture of anti mite bedding and household materials. Primary prevention will be based on better identification of populations at risk and on rationalisation of the alimentary diversification of infants, so controlling the environment of populations at risk. Finally, better understanding of the complex interaction of cytokines will allow consideration of creation, following the recent example of antileukotrienes, of new products of intervention and in particular regulation of the TH1-TH2 balance. PMID- 10370727 TI - The inordinate complexity of delivering laboratory services in the USA. PMID- 10370728 TI - Capillary electrophoresis in clinical chemistry. PMID- 10370730 TI - Review of investigation and management of severe hyponatraemia in a hospital population. AB - The purpose of this study was to assess retrospectively the prevalence of severe hyponatraemia in a hospital population and its laboratory investigation, treatment and clinical outcome. Over a 6-month period 47 patients (27 women and 20 men) were found to have a plasma sodium concentration of < or = 120 mmol/L (this number made up less than 0.17% of all plasma sodium requests over that time period). The mean patient age was 75 +/- 16 years and the average hospital stay was 37 +/- 45 days (1-179 days). Patient mortality was 51% (women 57% and men 43%). The mean initial plasma sodium concentration was 116 +/- 4.5 mmol/L, rising after therapeutic intervention to a mean of 130 +/- 4.2 mmol/L. The mean plasma sodium correction rate was 4.7 +/- 4.3 mmol/L/24 h (0.9-17.5 range). Twelve per cent of the patients had their plasma sodium raised at a rate of greater than 10 mmol/L/24 h after their initial presentation. Two patients may have had symptoms and signs suggestive of cerebral oedema/cortical dysfunction: in one patient the sodium concentration was raised at a rate of 9.5 mmol/L/24 h and in the other at 12.0 mmol/L/24 h. Sixty-one per cent of the patients had a chest infection, 44% were on diuretics, 28% had congestive cardiac failure, 28% were post-operative (9% orthopaedic procedures), 19% had carcinoma and 9% were on a selective serotonin re-uptake inhibitor. Regarding laboratory investigations, 56% had liver function tests, 41% had thyroid function tests, 36% had plasma osmolality determination, 36% had urinary electrolytes including urinary osmolality and < 2% had tests to exclude hypoadrenalism. PMID- 10370731 TI - Prevention of protein adsorption: effect on patient results for microalbuminuria. AB - Adding Triton X-100 to the diluent used for preparation of standard curves for protein assays can prevent non-specific adsorption of proteins from standards with a low protein content. Since results for low urinary albumin concentrations in control samples from different laboratories showed a wide range of mean values, we studied the effect of adding Triton X-100 to the diluent used in the preparation of the standard curve. Adding Triton X-100 resulted in absorbances which were more reproducible for both serum-based and purified-albumin-based standards. Results for control samples were also less variable in the Triton X 100 assay system. In control samples and patient samples the concentration of albumin was significantly lower in the Triton-X-100-based assay system. Adsorption of proteins from standards can only account for a small proportion of the variation in the range of mean results for urinary albumin in control samples from different laboratories. PMID- 10370729 TI - Assessment of corticosteroid replacement therapy in adults with adrenal insufficiency. AB - Recent work has taught us that our conventional approach to corticosteroid replacement therapy requires review. Specifically, the doses of hydrocortisone we have used are probably too high for the majority, and should ideally be administered in three or more doses through the day. Nevertheless, there is not much hard evidence that excessive glucocorticoid replacement per se will lead to adverse effects such as osteoporosis, even though it may exacerbate any tendency in those who are predisposed to it for other reasons. As such, there is no compelling need for using determinations of either UFC excretion or of the serum cortisol profile in the routine management of patients on replacement therapy. Nevertheless, such measures may be considered in those thought to be at particular risk of osteoporosis, and in whom it is felt that special effort should be made to ensure that they are receiving the minimum dose possible. In such circumstances, a cortisol day curve is likely to be of more value than measurement of UFC. PMID- 10370732 TI - Evaluation of five different high-density lipoprotein cholesterol assays: the most precise are not the most accurate. AB - We evaluated the accuracy and performance of four different test kits for the direct determination of high-density lipoprotein (HDL)-cholesterol and compared them with the phosphotungstic acid/MgCl2 assay. All four homogeneous assays were precise (within-run CV of < 2.0% and between-run CV of < 6.4%); both assays based on immuno-inhibition had the lowest CVs (within-run 1.3% and 0.9%; between-run 2.3% and 2.2%). Interference from haemolysis was negligible, but triglyceride concentrations gave a negative interference. The effects of conjugated and unconjugated bilirubin were opposite; conjugated bilirubin showed a negative interference of up to 40%; unconjugated bilirubin interfered positively up to 50%. Using the recently validated indirect phosphotungstic acid/MgCl2 method as a comparison, all four homogeneous assays did not fulfil the National Cholesterol Education Program total error standard, mainly due to the positive biases of 12 to 42%, apparently associated with improper calibrators. Both assays involving immuno-inhibition showed a concentration-dependent bias. PMID- 10370733 TI - Blood dolichols in a patient with abetalipoproteinaemia. AB - Dolichol and dolichyl derivatives have an important function as glycosyl carriers in the assembly of the N-asparaginyl-linked oligosaccharide core region of glycoproteins. Dolichols are synthesized through the cholesterol biosynthesis pathways in all mammalian organs and are present in all tissues, and are also associated with lipoproteins in the blood circulation. However, the origin and metabolic pathway of blood dolichols remain unknown. Abetalipoproteinaemia is a disorder of the secretion of very low-density lipoprotein (VLDL) from the liver and of chylomicrons from the intestine into the blood circulation. Therefore, examination of blood dolichols in this disorder may provide valuable information on their origin and metabolic pathway. Dolichols were exclusively associated with the high-density lipoprotein (HDL) fraction (80.7 +/- 6.3% of total dolichols) in control human blood. Serum from a patient also contained dolichols in the HDL fraction (82.8% of total dolichols). The total amount of dolichols was higher in the patient (207.0 ng/mL) than in the controls (106.2 +/- 22.7 ng/mL, n = 14). The compositions of dolichols were very similar to each other. These results indicated that, at least in the patient with abetalipoproteinaemia, the HDL associated dolichols were possibly derived from the liver not through other lipoproteins but through dolichol transfer protein, or were possibly taken up and carried by HDL from peripheral tissues. PMID- 10370734 TI - Influence of mammary artery as a bypass vessel on the results of seven biochemical assays after coronary artery bypass surgery. AB - We compared the changes in troponin T, creatine MB isoenzyme mass concentration (CK-MB mass), creatine kinase MB isoenzyme activity (CK-MB activity), creatine kinase (CK), alpha-hydroxybutyrate dehydrogenase (HBD), lactate dehydrogenase (LD) and aspartate aminotransferase (AST) concentrations after coronary artery grafting with saphenous vein grafts, without or in combination with uni- or bilateral internal mammary artery(ies) as bypass vessels in 73 patients. An increase in CK concentration after surgery was highest for the bilateral internal mammary artery bypass patient group and lowest for the group who received only saphenous vein grafts. We present 90th percentile values for the seven tests. PMID- 10370735 TI - Different strategies in the laboratory diagnosis of autoimmune disease: immunofluorescence, enzyme-linked immunosorbent assay or both? AB - We investigated the clinical utility of different strategies for antinuclear antibodies (ANA) and antibodies to extractable nuclear antigens (ENA) testing. All requests for ANA and ENA (n = 485) in a 20-week period were tested by immunofluorescence (FANA) and immunodiffusion (strategy 1), enzyme-linked immunosorbent assay (ELISA) techniques (strategy 2) or a combination of FANA and ELISA (strategy 3). Results of strategy 1 were positive by FANA in 8% (by immunodiffusion in 2%). By ELISA, 11% of the samples tested positive. In 12% (n = 60) of the cases the two strategies did not agree. The positive predictive value (PPV) for autoimmune disease of strategy 1 was significantly higher than that for strategy 2, but after exclusion of rheumatoid arthritis this difference was abolished. In strategy 2 reagent costs were high but working time comparably shorter. With strategy 3 PPV results were not better, whereas costs and working time were higher. The most frequently occurring reasons for ANA/ENA test requests were: joint symptoms (37%), follow up (30%) or abnormal laboratory result (7%). In a survey of the clinicians 66% replied that the test result did not have any consequences, irrespective of the result or the strategy used. We conclude that FANA and immunodiffusion are superior to ELISA techniques. However, the clinical value of ANA/ENA testing is low and more selective test ordering is strongly recommended. PMID- 10370736 TI - Three-colour, one-tube flow cytometric method for enumeration of CD4+ and CD8+ lymphocytes. AB - We investigated the usefulness of a one-tube, three-colour flow cytometric method for enumerating CD4+ and CD8+ lymphocytes. This method does not use any control antibodies and we compared it to the standard methods using either control and CD14/CD45 antibodies or control antibodies only on 38 blood samples from healthy and human immunodeficiency virus-infected patients. The one-tube method showed good agreement with the other, more complicated methods and is therefore suitable for reliable enumeration of CD4+ and CD8+ T-lymphocytes. PMID- 10370737 TI - Measurement of urinary amino acids using high-performance liquid chromatography equipped with a strong cation exchange resin pre-column. AB - We determined the urinary amino acid concentrations by high-performance liquid chromatography (HPLC) using an eluent buffer which had been processed through a column packed with strong cation-exchange resin. With this column, ghost peaks resolved or much reduced in size and shifted to a post-arginine position. A clearer separation of peaks of urinary amino acids and ammonia was obtained. This could be due to the elimination of O-phthalaldehyde reactive substances in the eluent by the resin column. This method does not require a separate step to remove ammonia, unlike most of the methods currently used, and thus saves time. PMID- 10370739 TI - Variations in serum angiotensin-converting enzyme activity following lung resection: a controlled study in a clinical setting. AB - Angiotensin-converting enzyme (ACE) plays an important role in post-operative cardiovascular control, especially in critical illness. Since the pulmonary endothelium is the major site of ACE production it would seem probable that surgical resection of lung tissue would significantly influence serum ACE (sACE) activity. The aim of this study was to investigate the effect of surgery on early post-operative sACE activity in patients undergoing lung resection (n = 18) and a control group of patients (n = 18) undergoing thoracotomy for other reasons. An early post-operative, sustained sACE fall without significant difference in sACE activity between the two groups was observed 6 h after the operation. Furthermore, there was no correlation between post-operative sACE variations and the amount of lung tissue resected. It appears that surgical removal of lung tissue does not significantly affect the post-operative sACE response. This may be due to the presence of important extra-pulmonary ACE-producing sites or to the compensation of the remaining pulmonary vascular endothelium. PMID- 10370738 TI - Measurement of phenylalanine and tyrosine in plasma by high-performance liquid chromatography using the inherent fluorescence of aromatic amino acids. AB - An isocratic high-performance liquid chromatography (HPLC) method is described using the natural fluorescence of phenylalanine and tyrosine compared with that of an internal standard N-methyl phenylalanine. Plasma precipitated with 6% perchloric acid was separated isocratically using a base-deactivated C18 column with 5% acetonitrile in water as the mobile phase. Fluorescent measurements at an excitation wavelength of 215 nm and emission 283 nm showed only three peaks for tyrosine, phenylalanine and the internal standard eluting within 9 min. Inter batch coefficients of variation for phenylalanine were 2.9% and 1.8% at levels of 70 and 567 mumol/L, respectively, and 2.9% at a level of 63 mumol/L for tyrosine. The results for phenylalanine for this method showed a small mean positive bias (11 mumol/L) when compared with the target all-method means for UK National External Quality Assessment Scheme samples (n = 31). The results for tyrosine showed a small positive mean bias (10 mumol/L) when compared with an ion-exchange chromatographic method (n = 40). This method provides a quick and simple alternative to those using HPLC with pre- or post-column derivatization for monitoring patients with phenylketonuria. It is also less subject to interferences than HPLC methods using ultraviolet detection, particularly for the early eluting tyrosine peak. PMID- 10370740 TI - Effect of oral supplementation with vitamin E on the oxido-reductive status of red blood cells in normal mice and mice subject to oxidative stress by chronic administration of adriamycin. AB - The effect of oral vitamin E supplementation on the oxido-reductive status of red blood cells in normal mice and those subject to oxidative stress by chronic administration of the anti-tumour drug Adriamycin was investigated. Mice were randomly separated into three groups of 20 animals each and maintained on diets identical in all respects except for vitamin E content. Group 1 received a low vitamin E diet that provided 10 mg vitamin E/kg body weight/day, group 2 received a normal mice chow diet (45 mg vitamin E/kg body weight/day) while group 3 received a high vitamin E diet (200 mg vitamin E/kg body weight/day). In comparison with the normal mice in group 1, their counterparts in groups 2 and 3 exhibited significantly higher (P < 0.001) activities of superoxide dismutase (SOD) in red blood cells (79.4% higher in group 2 and 114.2% higher in group 3, respectively) and produced lower concentrations of malondialdehyde (MDA) (22.9% less in group 2 and 51.2% less in group 3, respectively), with little difference in the glutathione peroxidase (GPX) activity. In Adriamycin-treated animals on the low vitamin E diet (group 1) the red blood cell SOD activity and MDA production were 46.2% and 200.7% higher (P < 0.001), respectively, and the GPX activity was 39.1% lower than in the red blood cells of untreated (normal) animals in the same group. The Adriamycin-induced changes were significantly less in animals receiving higher doses of vitamin E (groups 2 and 3). Thus, in the group maintained on the high vitamin E diet (group 3), Adriamycin administration resulted in only a 38.9% increase in the MDA production above that generated by red blood cells of normal mice in the same group, with no significant change in the SOD or GPX activities. Thus, in normal conditions as well as in conditions of oxidative stress, high doses of vitamin E appear to be able to protect the oxido reductive status of red blood cells by modulating the extent of lipid peroxidation as well as the activities of antioxidant enzymes. PMID- 10370741 TI - Behaviour of urinary dipeptidase in patients with chronic renal failure. AB - Renal dipeptidase (EC 3.4.13.19) activity in serum and urine from healthy volunteers (n = 20), patients with diabetes (n = 18) and patients with chronic renal failure (n = 5) was measured using glycyl-D-alanine as substrate. The assay was highly specific for the enzyme and was not affected by the various aminopeptidases present in serum and urine. No difference in serum renal dipeptidase activity was observed between the groups. The enzyme activity (U/L) in urine was higher than that in serum, irrespective of the group, suggesting the urine concentration was not affected by the serum concentration. The mean renal dipeptidase activities in urine were 2.56, 2.46 and 0.78 U/mol creatinine for healthy subjects, patients with diabetes and patients with chronic renal failure, respectively. The renal dipeptidase activity was significantly lower in the chronic renal failure group. The urinary excretion of dipeptidase (U/mmol creatinine) showed significant inverse correlations with that of beta 2 microglobulin, albumin and alpha 1-microglobulin, and with serum concentrations of creatinine, beta 2-microglobulin and alpha 1-microglobulin. We suggest that urine dipeptidase may be a useful marker of renal diseases. PMID- 10370742 TI - A kinetic model of mitochondrial aspartate aminotransferase transmigration in hepatobiliary disorders. AB - A hypothetical kinetic model of transmigration of mitochondrial aspartate aminotransferase (m-AST) from liver to blood and its elimination from blood was constructed and assessed for prediction by computer simulation. The elimination of m-AST from plasma in healthy rats followed first-order kinetics. Depletion of m-AST from the liver after induction of hepatobiliary disorders in rats with alpha-naphthylisothiocyanate (ANIT) also followed first-order kinetics. In contrast, the changes in metabolic rate in the liver after ANIT administration, as estimated from the plasma indocyanine green clearance, approximated to a second-order time function. Based on these data, the time-dependent change in plasma m-AST activity after ANIT administration was simulated by computer according to a hypothetical kinetic model. The result of computer simulation of the change in plasma m-AST coincided well with that obtained experimentally, suggesting that continuous measurements of m-AST may be helpful to the clinical diagnosis of liver injury. PMID- 10370743 TI - Measurement of glycated haemoglobin by boronate-affinity high-pressure liquid chromatography. PMID- 10370744 TI - Establishing trimester- and ethnic group-related reference ranges for fructosamine and HbA1c in non-diabetic pregnant women. PMID- 10370745 TI - Measurement of HbA1c by high-performance liquid chromatography in patients with renal failure. PMID- 10370746 TI - Effect of atorvastatin on low-density lipoprotein subfraction profile. PMID- 10370747 TI - Comparison of two methods for measurement of cardiac troponin T in patients with acute myocardial infarction. PMID- 10370748 TI - Serum retinol and alpha-tocopherol levels in healthy Asian subjects and patients. PMID- 10370749 TI - Serum chromogranin A and its relationship to endogenous markers of glomerular filtration rate. PMID- 10370750 TI - Detection of lactate dehydrogenase isoenzyme-M in body fluids containing albumin. PMID- 10370751 TI - Methionine loading in a Down's syndrome patient with cerebral infarction. PMID- 10370752 TI - Pseudocholinesterase deficiency: a dangerous, unrecognized complication of the ovarian hyperstimulation syndrome. PMID- 10370753 TI - Analysis of hCG: clinical applications and assay requirements. PMID- 10370754 TI - Influence of the laboratory upon requests for oral glucose tolerance tests for the diagnosis of diabetes mellitus. PMID- 10370755 TI - Paying for health care in the USA. PMID- 10370756 TI - New antiepileptic drugs. PMID- 10370757 TI - Carbohydrate-deficient glycoprotein syndromes: inborn errors of protein glycosylation. AB - The carbohydrate-deficient glycoprotein (CDG) syndromes (CDGS) are a series of autosomal recessive enzyme deficiencies which result in incomplete glycosylation of plasma proteins. CDGS types Ia and Ib have been related to deficiencies of phosphomannomutase and phosphomannose isomerase, respectively, while CDGS type II results from a deficiency of N-acetylglucosaminyltransferase II. Secondary CDG syndromes are associated with galactosaemia and hereditary fructose intolerance. The diagnosis of CDGS is most easily made by studying the glycoforms of suitable marker proteins using either electrophoresis or isoelectric focusing. This paper reviews the structure of the glycan chains of proteins and structural alterations in CDGS. It also outlines analytical techniques which are useful in the laboratory study of protein glycoforms and the diagnosis of CDGS. PMID- 10370758 TI - Hypophosphataemia in general practice patients. AB - We compared plasma phosphate concentrations in general practice patients and hospital inpatients and outpatients over an 8-month period. The distribution of results in all three groups was similar and 12-16% of results were at or below 0.8 mmol/L. In general practice patients, 8.3% of results from males and 12.1% from females were below the lower limit of their respective reference ranges. Eighteen of these patients (0.2% of results) had plasma phosphate concentrations < or = 0.4 mmol/L. On follow-up, only two of these patients had any attributable cause for their severe hypophosphataemia; in the remainder, it was unexpected and unexplained. Hypophosphataemia in outpatients and general practice patients is more common than has previously been appreciated. We present a strategy for further investigation of these patients. PMID- 10370759 TI - Levels of 24,25-dihydroxyvitamin D3, 25-hydroxyvitamin D3 and 25-hydroxyvitamin D3 3-sulphate in human plasma. AB - The concentrations of (24R)-24,25-dihydroxyvitamin D3[24,25(OH)2D3], 25 hydroxyvitamin D3[25(OH)D3] and its 3-sulphate [25(OH)D3(3)S] in the plasma of healthy subjects, patients with chronic renal failure, patients with climacteric syndrome, pregnant women and foetuses were determined using the enzyme-linked immunosorbent assay and high-performance liquid chromatography. 25(OH)D3(3)S was not detected in about one-third of the plasma samples from patients with chronic renal failure (n = 26). The three metabolites in maternal plasma reached the highest levels in the second trimester of pregnancy followed by a decrease to the values obtained in the first trimester. Older healthy women (age range 44-71 years) showed higher levels of 24,25(OH)2D3 and 25(OH)D3 in the plasma than did young healthy women (age range 21-29 years), whereas no clear difference was observed between the older healthy women and patients with climacteric syndrome. The level of 25(OH)D3(3)S in the plasma was higher in the latter patients than in healthy women. PMID- 10370760 TI - Influence of acetate and bicarbonate dialysate on blood short- and long-chain acyl carnitine in adult pyelonephritis patients. AB - The effect of acetate and bicarbonate haemodialysis (HD) on the concentrations of erythrocyte, whole-blood and plasma total carnitine (TC), free carnitine (FC), short- (SC) and long-chain acylcarnitine (LC) and acylcarnitine (AC) as well as the ratio of AC to FC was investigated in 30 healthy subjects (15 men and 15 women) and 27 patients (10 men and 17 women) with chronic pyelonephritis (CPN) undergoing chronic HD. Fourteen patients (5 men and 9 women) used acetate HD and the remainder (5 men and 8 women) used bicarbonate HD. The mean predialysis erythrocyte, whole-blood and plasma concentrations of TC, FC, SC, LC and AC as well as the ratio of AC to FC were not significantly different from those in healthy controls (P > 0.05). However, after acetate or bicarbonate HD, a significant decrease in erythrocyte, whole-blood and plasma concentrations of TC, FC, SC, LC and AC were found, compared with either predialysis or healthy control values (P < 0.001). Furthermore, the ratio of AC to FC was significantly higher following acetate HD as compared with either acetate or bicarbonate predialysis values (P < 0.001). The observed variations in response between acetate and bicarbonate HD may be due to enhanced formation of acetyl-coenzyme A and fatty acid synthesis during acetate HD. PMID- 10370761 TI - Biological variability and reference intervals for total plasma homocysteine. AB - We determined the intra-individual biological variability of plasma homocysteine in 20 healthy subjects. The intra-individual coefficient of variation was relatively low (8.3%), indicating that a single measurement can be used to characterize the average homocysteine concentration. A population study measuring plasma homocysteine and serum folate levels was conducted on serum samples collected from 1109 randomly selected, fasting adults with a wide age range. We determined age- and gender-specific central 0.95 intervals and found that subjects in the highest quartile of serum folate had significantly lower (P = 0.0001) mean plasma homocysteine concentrations than did those in the lowest quartile of folate values. An 'ideal' homocysteine reference range, based on targeting those subjects who are likely to be folate replete, is preferable to the population-based range using the central 0.95 interval. PMID- 10370762 TI - Essential fatty acids profile and lipid peroxides in severe pre-eclampsia. AB - There is increasing evidence that an imbalance between products of arachidonic acid metabolism (such as prostaglandins, prostacyclin and thromboxane A2) and lipid peroxidation products might have a role in the pathogenesis of pre eclampsia. In order to study some of the changes taking place in pre-eclampsia, the plasma concentrations of essential fatty acids and lipid peroxidation products [measured as thiobarbituric-acid (TBA)-reactive substance] were evaluated in 25 patients with severe pre-eclampsia and 20 normal pregnant control subjects. Arachidonic acid concentration (median and range) was significantly higher in the pre-eclamptic group (14.1; 11.0-19.8 micrograms/L) compared with the normal pregnant subjects (2.5; 2.0-4.4 micrograms/L). Plasma concentration of TBA-reactive substance was also significantly higher in the pre-eclamptic cases (14.8; 13.4-16.8 vs 11.4; 9.0-12.8). These results suggest that arachidonic acid and lipid peroxidation products may play a role in the pathogenesis of pre eclampsia. PMID- 10370763 TI - The portable laboratory: an evaluation of the accuracy and reproducibility of i STAT. AB - The i-STAT point-of-care testing analyser, which utilizes single-use multisensor cartridges, was evaluated. Two cartridges were assessed: the 6+ and the G3+, which provide results for urea, glucose, sodium, potassium, chloride, haematocrit, pH, PCO2, PO2 and various calculated parameters. The results for all analytes agreed well with the analysers in routine use in the laboratory. The reproducibility was comparable even when analysis was carried out by a nurse with only 5 min training. The system was found to be reliable, easy to use and required no maintenance (only a 2-min daily check of the electronics). These features, together with portability and the storage capacity for results, make the i-STAT suitable for point-of-care use, particularly in critical care units. PMID- 10370764 TI - Analytical error of home glucose monitors: a comparison of 18 systems. AB - In a quality review of 18 home blood glucose monitors, we measured the imprecision and incidence of significant error of 12 colorimetric and six amperometric systems by using capillary blood specimens from patients attending diabetes clinics. Imprecision at the mean glucose concentration found in the respective blood specimens (about 9 mmol/L) gave coefficients of variation (CV) ranging from 5.2 to 22.8%. Eight monitors including five of amperometric design had a CV of less than 10%. The incidence of significant error (defined as the proportion of specimens differing in value by more than 15% from a reference hexokinase assay of glucose in capillary blood) varied from 6 to 76%. Among the eight monitors identified as being most precise, the majority produced results that differed markedly from the reference assay, underlining the need for a common approach to calibration of home glucose monitors. PMID- 10370765 TI - Survey of serum potassium reference measurements. AB - We compared the quality of reference measurements for serum potassium in four reference laboratories from three different European countries, using a panel of 60 native patients' samples. The reference methods were based on either ion chromatography (one laboratory) or flame atomic emission spectrometry (three laboratories). Performance specifications for serum potassium measurements were defined as a maximum overall coefficient of variation (CV) of 1.5%, a maximum bias of 0.65% and a maximum total error of 3.0%. The overall imprecision for all laboratories was in the range of 0.7 to 1.3%, and was thus below the proposed specification of 1.5%. However, two laboratories reported 12 and 13 quadruplicates with CVs exceeding this limit. The mean bias (expressed as deviation from the overall mean of all laboratories) for all reference laboratories was < 0.65%. In the lower concentration range, however, one laboratory exceeded this limit. No laboratory measured samples with a total error above 3.0%. From these results, it can be concluded that the reference measurements, and, thus, also the reference methodologies, based on ion chromatography and flame atomic emission spectrometry were equivalent, and able to satisfy current analytical specifications for serum potassium measurements. PMID- 10370766 TI - Serum and tissue antioxidant capacity in cervical intraepithelial neoplasia investigated using an enhanced chemiluminescent reaction. AB - Depleted antioxidant defence has been implicated in the pathogenesis of cervical neoplasia. We determined the systemic and local antioxidant status of women with this condition. Thirty-four women with varying grades of cervical intraepithelial neoplasia, 25 patients who had been treated successfully with diathermy loop excision, and 56 women who had no evidence of cervical abnormality acted as controls. Total antioxidant capacity of serum and protein-free serum, and of neat and protein-free homogenized cervical punch biopsies were determined using enhanced chemiluminescence. Mean serum antioxidant capacity of patients with current neoplasia and treated patients was not significantly different from that of controls. However, mean antioxidant capacity of homogenized cervical tissue from women with neoplasia was significantly lower than control means (P < 0.005), while results for treated patients were intermediate between those from diseased and normal samples (P < 0.05). The enhanced chemiluminescence technique has potential as a suitable method for measuring total antioxidant capacity of cervical tissue, and warrants further investigations using other tissue types. Significant antioxidant depletion in cervical intra-epithelial neoplasia appears to be confined to the local cervical mucosa. PMID- 10370767 TI - Assessment of the feasibility of conducting population prevalence studies of chronic renal failure according to ethnic group: a survey of clinical biochemistry laboratories in Greater London and south east England. AB - The planning of renal replacement therapy is based on assessment of population need for the white population, but uses historical trends in treatment uptake for black and Asian ethnic minority groups, for whom the incidence of chronic renal failure (CRF) is not known. Epidemiological studies of CRF are based upon follow up of plasma creatinine results obtained from clinical biochemistry laboratories. We conducted a postal questionnaire survey of UK National Health Service (NHS) and private clinical biochemistry laboratories in Greater London and the south east of England to arrange the design and test the feasibility of carrying out a study to determine ethnic-specific rates of CRF. Fifty-five NHS laboratories (90%) and 19 private laboratories (57%) responded. Few pathology computer systems recorded ethnic group, patient post code, or diagnosis; although 31 of the laboratory computers (42%) were linked with the hospital Patient Administration System which could supply these data. Approximately 5.5 million electrolyte profiles and 20 million individual renal function tests are carried out annually in south east England. Ninety per cent of those were performed within NHS laboratories, implying that a study can use NHS sources alone without risk of any undue bias. Sixty laboratories (81%) included creatinine in their routine electrolyte profile, which would be a requirement for any study. Thirty-one laboratories (42%) archived tests within 1 year of entry, which would rule out a retrospective study design. A prospective study is feasible and should be carried out as soon as is practicable. PMID- 10370768 TI - Problematic performance of thyroid stimulating hormone kit in external quality assessment does not affect its popularity. PMID- 10370769 TI - Accuracy of Down's syndrome risks produced in a prenatal screening program. AB - Down's syndrome screening in the second trimester using a variety of combinations of maternal serum markers has become an established part of antenatal care over the past decade. The methodology of calculating Down's syndrome risk using statistical models based on the Gaussian distribution functions of the various markers used is often considered daunting, and many screening centres rely on specific screening computer software to undertake the relevant calculations. Such software is often a closed system which does not allow the user to check or change any of the parameters within the model. It is clear from a recent UK NEQAS survey of Down's syndrome screening laboratories (Spencer, Ellis, Seth, 1997, unpublished data) that many centres are unsure what method of calculating risk and what population parameters are used in their software, and how appropriate these are for the population being screened. This confusion has led to uncertainty over the reliability of the risk estimates produced in Down's syndrome screening programs. PMID- 10370770 TI - Total antioxidant power of plasma: male-female differences and effect of anticoagulant used. PMID- 10370771 TI - Raised urinary pyridinium cross-links in humoral hypercalcaemia of malignancy. PMID- 10370772 TI - Caution is needed in point-of-care monitoring of oxygen therapy in chronic obstructive pulmonary disease. PMID- 10370773 TI - A survey of endocrine function testing by clinical biochemistry laboratories in the UK. PMID- 10370774 TI - External quality assurance--a personal view. PMID- 10370775 TI - Secondary adrenal failure in a young woman presenting as hypoglycaemic coma. PMID- 10370776 TI - Rationing in cancer treatment: is the identification of essential cytotoxic drugs feasible? PMID- 10370777 TI - Drug combinations: dangerous liaisons or great expectations? PMID- 10370779 TI - Essential drugs for cancer therapy: a World Health Organization consultation. AB - The WHO has previously produced recommendations on the essential drugs required for cancer therapy. Over the last five years several new anti cancer drugs have been aggressively marketed. Most of these are costly and produce only limited benefits. We have divided currently available anti-cancer drugs into three priority groups. Curable cancers and those cancers where the cost-benefit ratio clearly favours drug treatment can be managed appropriately with regimens based on only 17 drugs. All of these are available, at relatively low cost, as generic preparations. The wide availability of these drugs should be the first priority. The second group of drugs may have some advantages in certain clinical situations. Based on current evidence, drugs in the third group are judged as currently not essential for the effective delivery of cancer care. Adequate supportive care programmes with the widespread availability of effective drugs for pain control are of considerably greater importance. The adoption of these priorities will help to optimise the effectiveness and efficiency of chemotherapy and ensure equitable access to essential drugs especially in low resource environments. Clearly this paper represents the views of its contributors. The WHO welcomes feedback from all oncologists so that the advice it gives to governments in prioritising the procurement of anti cancer drugs can be as comprehensive as possible. PMID- 10370778 TI - The third-generation non-steroidal aromatase inhibitors: a review of their clinical benefits in the second-line hormonal treatment of advanced breast cancer. AB - Three new aromatase inhibitors have recently completed phase III evaluation as treatment of metastatic breast cancer in post-menopausal women whose disease has progressed despite tamoxifen therapy: anastrozole (ARIMIDEX, Zeneca), letrozole (FEMARA, Novartis) and vorozole (RIVIZOR, Janssen). All belong to the third generation of non-steroidal aromatase inhibitors, and each is superior to previous generations in terms of potency and selectivity. The trials that have been performed compare each agent to megestrol acetate, and letrozole and vorozole to aminoglutethimide. Although the studies are not directly comparable due to differing study designs and patient populations, it has been demonstrated each of these drugs provides single agent, once-daily, oral palliation of hormone responsive, post-menopausal metastatic breast cancer. Letrozole is clearly more effective than megestrol acetate, and anastrozole and vorozole are possibly so. All three are better tolerated than the progestin, particularly in terms of weight gain. Both letrozole and vorozole are significantly more effective, and better tolerated than aminoglutethimide. Overall, this most recent generation of aromatase inhibitors is a clear improvement on our current standard second-line therapies. In 1999, tamoxifen remains the first choice in the hormonal therapy of breast cancer. Following tamoxifen failure, the optimal second-line hormonal therapy remains undefined, but aminoglutethimide and megestrol acetate are no longer optimal therapy in this setting. The third-generation non-steroidal aromatase inhibitors must now be compared to each other, to the steroidal aromatase inhibitors, to the pure anti-oestrogens, and to tamoxifen. PMID- 10370780 TI - Pharmacokinetic interactions of paclitaxel, docetaxel and their vehicles with doxorubicin. AB - BACKGROUND: The combination of doxorubicin (Dx) with paclitaxel or docetaxel is clinically effective but there are concerns regarding the higher incidence of cardiotoxicity of the combination compared with Dx alone. The mechanism of the increased toxicity is still unclear. PURPOSE: To assess whether there is a pharmacokinetic interaction between paclitaxel, docetaxel or their vehicles and Dx in mice. MATERIALS AND METHODS: CDF1 male mice were treated with Dx either alone (10 mg/kg i.v.) or in combination with paclitaxel (25 mg/kg) or docetaxel (25 mg/kg) or their vehicles, i.e., cremophor-ethanol-glucose (cremophor) or polysorbate80-ethanol-glucose (polysorbate). Four mice were killed 4, 8 or 24 hours after Dx in each experimental group and Dx was assayed in serum and in heart, liver, kidney and spleen by HPLC. RESULTS: Four hours after treatment the concentrations of Dx in heart, liver and kidney were much higher in mice concomitantly treated with paclitaxel, docetaxel (dissolved in either cremophor or polysorbate) and cremophor. At subsequent times the differences were modest and only reached statistical significance in a few cases. Dx metabolites were modified by concomitant treatment with taxanes or their vehicles. In particular, the levels of Dx aglycone in liver and kidney were significantly lower in mice treated with the combination than in mice given Dx alone. CONCLUSIONS: Paclitaxel, docetaxel and cremophor when given together with Dx modify its distribution and metabolism, increasing Dx levels in many tissues including the heart. This might have some bearing on the toxicity of regimens in which Dx is combined with taxanes. PMID- 10370781 TI - Vinorelbine as first-line chemotherapy for advanced breast cancer in women 60 years of age or older. AB - BACKGROUND: Older patients with advanced breast cancer are less likely to receive chemotherapy than younger patients. Vinorelbine is an attractive alternative in this setting because of its clinical activity and low frequency of side effects. This multicenter, phase II trial was designed to assess the safety and efficacy of intravenous vinorelbine as first-line therapy in women > or = 60 years old. PATIENTS AND METHODS: Fifty-six women (median age, 72 years; range 60-84 years), with measurable advanced breast cancer and no prior chemotherapy for metastatic disease, were enrolled and included in the analysis. Vinorelbine 30 mg/m2 was administered weekly for 13 weeks and then every two weeks until development of progressive disease; doses were reduced or delayed to manage toxicity. RESULTS: The objective response rate was 38% (95% confidence interval (95% CI): 24%-51%); median duration of response, nine months; median time to disease progression in all patients, six months. The major dose-limiting toxicity was hematologic, which led to a median dose intensity of 20.6 mg/m2/week. Grade 3-4 nonhematologic toxicity consisted of asthenia (7%); nausea and generalized pain (5%); vomiting, chest pain, abdominal pain, and elevated AST (4%); fever, diarrhea, constipation, and injection site reaction (2%). Neurotoxicity and alopecia were grade 1-2 and relatively infrequent. CONCLUSIONS: Vinorelbine offers a promising alternative for the management of advanced breast cancer in elderly patients who are concerned about the subjective side effects of cytotoxic chemotherapy. The dose limiting toxicity is neutropenia, which is readily managed with dose adjustment. Nonhematologic toxicity, including gastrointestinal side effects, is minimal. Randomized studies are warranted to compare the activity of vinorelbine with that of other regimens in elderly patients. PMID- 10370782 TI - Paclitaxel by 24-hour infusion with doxorubicin by 48-hour infusion as initial therapy for metastatic breast cancer: phase I results. AB - PURPOSE: We and others have demonstrated the antineoplastic efficacy of paclitaxel as a single agent in metastatic breast cancer. We performed this phase I trial to evaluate the combination of paclitaxel with doxorubicin. PATIENTS AND METHODS: Eligible patients had measurable or evaluable metastatic breast cancer for which this was the initial cytotoxic treatment. They may have received adjuvant chemotherapy with other drugs. The study had four parts. In part 1, the patients received paclitaxel by 24-hour infusion followed by doxorubicin by 48 hour infusion. The paclitaxel dose was to be escalated from a starting dose of 125 mg/m2, and the doxorubicin dose was to remain constant at 60 mg/m2 with treatment repeated every three weeks. The results of part 1 prompted part 2 which was a study of the reverse sequence. Part 3 was a formal study of pharmacology and has been reported (J Clin Oncol 14: 2713-21, 1996). In part 4, patients received doxorubicin 50 mg/m2 by bolus followed by paclitaxel 150 mg/m2 by 24 hour infusion for courses 1 and 2. In all subsequent courses doxorubicin was administered by 48-hour infusion. All patients in all four parts of the study had baseline cardiac scans. All patients received standard premedication for paclitaxel. RESULTS: Forty-eight patients were treated in all four parts of the study. In part 1 (10 patients), the maximum tolerated dose (MTD) was paclitaxel 125 mg/m2/24 hours followed by doxorubicin 48 mg/m2/48 hours as defined by dose limiting mucositis and neutropenic fever which occurred at the starting dose. For part 2 (21 patients), the MTD was doxorubicin 60 mg/m2/48 hours followed by paclitaxel 160 mg/m2/24 hours. In part 4 (seven patients), the MTD was doxorubicin 50 mg/m2/bolus followed by paclitaxel 135 mg/m2/24 hours. In parts 2 and 4, the dose-limiting toxic effect was neutropenia. Of the entire cohort of 48 patients, seven (15%) had a complete response (one persists at five years without intervening therapy), 26 (54%) had a partial response for an objective response rate of 69% (95% confidence interval (95% CI): 54%-81%). The median follow-up of all living patients is 38+ months (range 20+ to 62+); the median response duration is seven months (range 2-33.7+); the median overall survival is 20.5 months (range 5-54+). The median time to progression is 9.6 months (range 1-33.7+ months). Two patients developed congestive heart failure, one at 24 months after her final dose of doxorubicin which amounted to a cumulative lifetime total doxorubicin dose of 870 mg/m2, one after a total of 660 mg/m2. In both, cardiac symptoms were controlled with medications. CONCLUSIONS: The combination of paclitaxel/24 hours with doxorubicin/48 hours is an effective antineoplastic treatment for metastatic breast cancer. However, the incidence of complete response, the median overall survival, and time to progression were not greater than for standard doxorubicin-based combinations. Additionally, a sequence dependent interaction between paclitaxel and doxorubicin, given in the schedule described here, was defined. Other strategies and schedules should be evaluated to maximize the antineoplastic efficacy of these two potent agents. PMID- 10370783 TI - Metaplastic breast cancer: prognosis and response to systemic therapy. AB - BACKGROUND: Metaplastic breast cancer is a rare disease with little information available to guide therapy. The goals of this study were to describe the patient characteristics, systemic therapies and clinical outcomes of all patients with primary metaplastic breast cancer treated at Mayo Clinic between 1976 and 1997. PATIENTS AND METHODS: Patients were identified through the medical index of Mayo Clinic. Clinical information was abstracted from the medical record of each patient. A literature search using MEDLINE and CANCERLIT for the years 1966-1997 was performed to identify all previously reported case series in the English language containing 10 or more patients. RESULTS: Twenty-seven patients were identified with a median age at diagnosis of 59 years (range 39-90 years). The median tumor size was 3.4 cm (range 0.5-7.0 cm). One patient had metastatic disease at presentation. Twenty-three patients had information available on nodal status, estrogen receptor (ER) and progesterone receptor (PR) status. Twenty patients (87%) were node-negative and three patients (13%) were both ER and PR positive. Disease-free survival (DFS) and overall survival (OS) were assessed for those who presented with local-regional disease. The three-year DFS was 40% (95% CI: 23%-73%) and the three-year OS was 71% (95% CI: 51%-97%). In univariate analysis, those patients 60 years of age or older at diagnosis were found to have an increased DFS (P = 0.011). Among those with prior estrogen use, both DFS (P = 0.022) and OS (P = 0.003) were decreased. Thirteen patients (50%) developed metastases with a median DFS time of 2.4 years. Ten different chemotherapy regimens were utilized for metastatic disease and one partial response was observed. There were no responses to tamoxifen in four patients with metastatic disease. Median survival after the development of metastases was eight months. CONCLUSIONS: Despite presenting more commonly as node-negative disease, DFS and OS in metaplastic breast cancer is decreased compared to typical adenocarcinomas. Systemic therapy also appears to be less effective. Patients with metaplastic breast cancer, particularly those with metastatic disease could be appropriate candidates for innovative therapeutic regimens. PMID- 10370784 TI - Phase II trial combining mitomycin with 5-fluorouracil, epirubicin, and cisplatin in recurrent and metastatic undifferentiated carcinoma of nasopharyngeal type. AB - BACKGROUND: This phase-II study was conducted to investigate the potential benefit from the addition of mitomycin to a conventional anthracycline-cisplatin- and 5-fluorouracil-based chemotherapy for recurrent and metastatic undifferentiated carcinoma of nasopharyngeal type (UCNT). PATIENTS AND METHODS: Between July 1989 and December 1991, 44 consecutive patients (M/F 36/8; median age: 45, range 20-72; performance status (PS) 0: 20 patients, PS 1: 14 patients, PS 2: 10 patients) with recurrent or metastatic UCNT were entered in this study after complete clinical, biological, and radiological pre-therapeutic work-ups. Chemotherapy (FMEP regimen) consisted of 800 mg/m2/day 5-fluorouracil in continuous infusion from day 1 to day 4 combined with 70 mg/m2 epirubicin, 10 mg/m2 mitomycin, and 100 mg/m2 cisplatin on day 1, every four weeks for six cycles. Mitomycin was delivered in cycles 1, 3, and 5 only. Eleven patients had isolated loco-regional recurrences, 12 patients had local recurrences associated with distant metastasis, and 21 patients had metastasis only. Toxicity and response were evaluated according to WHO criteria. TOXICITY: Grade 3-4 neutropenia was observed in 122 of 212 evaluable cycles (57%) and 39 of 44 patients (89%); febrile neutropenia occurred in 16 patients (36%) and 24 cycles (11.3%). Grade 3-4 thrombocytopenia was observed in 27 patients (61%) and 45 cycles (21%), including 27 of 45 cycles (60%) with mitomycin. Grade 3 anemia was noted in 18 patients (40%) and 23 cycles (11%), including 18 of 23 cycles (78%) with mitomycin. Grade 3-4 mucositis occurred in 25 cycles (11%) and 14 patients (32%), mainly in those previously treated with radiation therapy in the head and neck area. There were four treatment-related deaths (9%); three of them neutropenia-related, and one of cardiac toxicity. RESPONSE: Forty-four patients were evaluable for response: There were 23 of 44 objective responses (52%), including six complete responses (13%), and 17 partial responses (38%). Additional radiotherapy was given to 13 patients after documentation of response: Nasopharyngeal tumor + cervical nodes (eight patients) and/or on bone metastasis sites (five patients); mediastinal lymph nodes (one patient). At a median follow up of 87 months (range 71-100), five patients are alive and in continuous complete remission. The median survival time was 14 months and the median time to progression nine months. CONCLUSION: The regimen under study is active in recurrent/metastatic UCNT, but associated with excessive toxicity. PMID- 10370785 TI - Clinical outcome after autologous transplantation in non-Hodgkin's lymphoma patients with high international prognostic index (IPI). AB - BACKGROUND: Dose intensification and autologous stem cell transplantation as front-line therapy in non-Hodgkin's lymphoma patients (NHL) is a matter for debate, although preliminary data suggest a role for it in patients at high risk of resistance or relapse according to the international prognostic index (IPI). PURPOSE AND STUDY DESIGN: To compare retrospectively the clinical outcome of two cohorts of NHL patients with high-risk IPI treated with MACOP-B for 12 weeks (38 patients) or high-dose chemotherapy (44 patients) including eight weeks of MACOP B, one or two intensification cycles with mitoxanthrone, dexamethasone, high-dose ara-C and finally BEAM chemotherapy with autologous hemopoietic progenitor cell transplantation. RESULTS: The actuarial estimate of event (progression, relapse or death)-free survival (EFS) at three years was better (58% vs. 41%, P = 0.08) for patients treated with intensive regimen even though the overall survival did not show a statistically significant difference (63% vs. 50%, P = 0.27). Multivariate analysis showed that the high-dose chemotherapy program was the only independent variable correlating with a reduction in the event rate. CONCLUSION: Early autologous stem-cell transplantation might improve the clinical outcome of high-risk patients according to IPI. PMID- 10370786 TI - Interleukin-10 levels are often elevated in serum of adults with Hodgkin's disease and are associated with inferior failure-free survival. AB - BACKGROUND: Interleukin-10 (IL-10) is a pleiotropic cytokine that protects B- or T-lymphocytes and hemopoietic progenitors from apoptosis induced by doxorubicin, glucocorticoids, or deprivation of growth factors. IL-10 is also immunosupressive, and tumor cells secreting IL-10 can grow in syngeneic or allogeneic hosts, and can inhibit the generation of tumor-specific cytotoxic T cells. Hodgkin-Reed-Sternberg cells are derived from follicular center B cells and they may be latently infected by EBV. When this occurs they often express IL 10. Based on these considerations we investigated the relationship between pretreatment serum IL-10 levels and failure-free survival (FFS) in Hodgkin's disease (HD). PATIENTS AND METHODS: Untreated patients, older than 16 years, with biopsy-proven HD, were included if treated with ABVD or equivalent regimens, and if pretreatment serum was available. IL-10 levels were determined with a capture enzyme-linked immunoassay specific for cellular IL-10. RESULTS: Among healthy adult volunteers serum IL-10 levels ranged from 4.8-9.8 pg/ml (mean 7.1, standard deviation 1.5 pg/ml). Therefore levels > or = 10 pg/ml were considered elevated. We identified 101 patients with available serum. Their median age was 32 years, and 60% had B-symptoms. Ann Arbor stage was I in 4, II in 21, III in 35, and IV in 41 patients. Histology was nodular sclerosis in 74, mixed cellularity in 12, lymphocyte predominance in six, lymphocyte depletion in one, and unclassified in eight patients. Pretreatment serum IL-10 levels were elevated in 51 patients, and were higher in those with serum albumin < 3.5 g/dl, B symptoms, serum beta 2 microglobulin > or = 2.5 mg/l, anemia, and AAS III or IV. After a median follow up of 32 months for survivors, 20 patients have progressed, and the three-year FFS of those with high vs. normal serum IL-10 was 60% +/- 9 vs. 91 +/- 9% (50% vs. 50% of the population; P = 0.004 by log-rank). Among patients with Ann Arbor stage III or IV the three-year FFS for those with high vs. normal serum IL-10 (58 vs. 42% of the population) was 57 +/- 9% vs. 92 +/- 6% (P = 0.008 by log-rank). Multivariate analysis using Cox's proportional hazards model confirmed that IL-10 was an independent variable associated with inferior FFS in this population. CONCLUSIONS: Elevation of serum IL-10 levels is frequent and is associated with inferior FFS in adults with ABVD-treated HD. This observation should be verified in other patient populations. In addition, the source and the role of IL-10 in the biology of HD should be further investigated. PMID- 10370787 TI - Pharmacokinetic schedule finding study of the combination of gemcitabine and cisplatin in patients with solid tumors. AB - PURPOSE: To determine possible schedule dependent pharmacokinetic and pharmacodynamic interactions between gemcitabine (2',2'-difluorodeoxycytidine, dFdC) and cisplatin (cis-diammine-dichloroplatinum, CDDP) in patients with advanced stage solid tumors in a phase I trial. PATIENTS AND METHODS: A total of 33 patients with advanced stage solid tumors were treated with gemcitabine (30 min infusion, 800 mg/m2) and cisplatin (one-hour infusion, 50 mg/m2). Sixteen patients had a four-hour interval between gemcitabine (days 1, 8, 15) and cisplatin (days 1 and 8), followed by the reverse schedule and seventeen patients had a 24-hour interval between gemcitabine (days 1, 8, 15) and cisplatin (days 2 and 9), followed by the reverse schedule. Gemcitabine and cisplatin pharmacokinetics were measured in plasma and white blood cells (WBC), isolated from blood samples taken at several time points after the start of treatment. RESULTS: A four-hour time interval between both agents did not reveal major differences in plasma pharmacokinetics of gemcitabine, dFdU (deaminated gemcitabine) and platinum (Pt), and of gemcitabine-triphosphate (dFdCTP) accumulation and Pt-DNA adduct formation in WBC between the two different sequences of gemcitabine and cisplatin. In the patients treated with the 24-hour interval, cisplatin before gemcitabine did not significantly change peak gemcitabine levels and the AUC of plasma dFdU, but tended to increase dFdCTP AUC in WBC 1.5-fold (P < 0.06). Gemcitabine before cisplatin decreased the plasma AUC of Pt 2.1-fold (P = 0.03). No significant differences in Pt-DNA adduct levels in WBC were found, although gemcitabine before cisplatin tended to increase the 24 hour retention of Pt-DNA adducts. Creatinine clearance on day 28 was related to the peak plasma levels of total Pt (linear regression coefficient (r) = 0.47, P = 0.02, n = 26). Furthermore, the increase in the Pt-GG to Pt-AG ratio 24 hours after cisplatin treatment was related to the overall response of patients (r = 0.89, P < 0.01, n = 8). CONCLUSIONS: Of all schedules the treatment of patients with cisplatin 24 hours before gemcitabine led to the highest dFdCTP accumulation in WBC and total Pt levels in plasma. These characteristics formed the basis for further investigation of this schedule in a phase II clinical study. PMID- 10370788 TI - Activity of SCH 66336, a tricyclic farnesyltransferase inhibitor, against human tumor colony-forming units. AB - BACKGROUND: The ras gene product regulates transduction of growth-proliferative signals from the membrane to the nucleus. Mutationally-activated Ras is the oncogene most frequently found in human tumors. In order to perform its function in cell signaling, Ras must be farnesylated on the CAAX motif present on the carboxyl terminus of the ras protein. This reaction is catalysed by farnesyl protein transferase. In the present study, SCH 66336, an orally bioavailable nonpeptide tricyclic farnesyltransferase inhibitor, was tested against a large variety of human tumors to define its preclinical activity profile, utilizing the human tumor cloning assay. MATERIALS AND METHODS: A soft agar cloning assay was used to determine the in vitro effects of SCH 66336 against primary human tumor specimens taken directly from patients. A total of 70 evaluable specimens were exposed to SCH 66336 for 14-day continuous exposure at concentrations ranging from 0.1 to 2.5 microM. In vitro responses were defined as an inhibition > or = 50% of human tumor colony forming units at a given concentration. RESULTS: There was a positive relationship between concentration and response to SCH 66336. With the highest concentration (2.5 microM), response was demonstrated in 50% (three of six) of breast tumors, 40% (6 of 15) of ovarian tumors, and 38% (5 of 13) of non-small-cell lung tumor colony forming units. Among the 69 specimens tested at the concentration of 2.5 microM, SCH 66336 had activity in 27% of tumor specimens that were resistant to doxorubicin, 38% of tumor specimens resistant to cisplatin, 33% of tumor specimens resistant to paclitaxel, and 27% of tumor specimens resistant to etoposide. CONCLUSIONS: The broad spectrum of soft agar growth inhibition by SCH 66336 in the human tumor cloning assay, and its efficacy at physiologically relevant concentrations in animal models, suggest that SCH 66336 may deserve future clinical trials in patients with ovarian, breast and non small-cell lung cancer. PMID- 10370789 TI - Induction of gamma-glutamylcysteine synthetase gene expression by platinum drugs in peripheral mononuclear cells of lung cancer patients. AB - BACKGROUND: To investigate in vivo the roles of gamma-glutamylcysteine synthetase (gamma-GCS), multidrug resistance-associated protein (MRP), human canalicular multispecific organic anion transporter (cMOAT) and DNA topoisomerase I (topo I) in relation to platinum drug resistance, we monitored the changes of the steady state levels of the mRNAs for these factors in peripheral mononuclear cells (PMN) after completing platinum drug administration. PATIENTS AND METHODS: PMN from 46 subjects were studied. We obtained PMN from 14 previously untreated lung cancer patients and 14 normal volunteers to measure the baseline gene expression levels. We then obtained PMN from 18 patients with previously untreated advanced lung cancer before and after they received platinum drug treatment. We analyzed the gene expression levels by using the quantitative reverse transcription polymerase chain reaction (RT-PCR). RESULTS: There were no differences in the baseline expression levels between normal volunteers and lung cancer patients in any of the genes. After platinum drug administration, the heavy subunit of gamma-GCS (gamma-GCSh) expression level increased 2.5-fold within 24 hours and the increase persisted for a month, whereas the light subunit of gamma-GCS (gamma-GCSl) expression level did not show an early response but had increased after a month. By contrast, the MRP, cMOAT and topo I expression levels were similar before, during and after chemotherapy. CONCLUSIONS: These results suggest that the gene expression levels of both subunits of gamma-GCS play an important in vivo role in platinum drug resistance. PMID- 10370790 TI - Cytomegalovirus pneumonia in a patient with breast cancer on chemotherapy: case report and review of the literature. AB - Cytomegalovirus (CMV) pneumonia in the setting of non-transplantation patients is a rarity. We present a case of CMV pneumonitis in a woman with stage IV breast cancer, with brain metastases, receiving both chemotherapy and systemic corticosteroids. A review of the literature reveals this as a unique case. Potential viral etiologies should therefore be considered in cancer patients with pneumonia receiving non-transplantation chemotherapy-regimens, particularly if steroids are a component of their therapy. PMID- 10370791 TI - The oncogenic 30 and 69 bp deletion variants of the EBV LMP-1 gene are common in HIV-negative lymphoproliferations, both malignant and benign. AB - BACKGROUND: In vitro studies have shown that the 30 and 69 base pair (bp) deletion variants of the latent membrane protein (LMP)-1 gene of the Epstein-Barr virus (EBV) have a higher transforming capacity than the wild-type variant. In recent years these studies have triggered an in vivo search for such potentially oncogenic variants in lymphoid tissues. PATIENTS AND METHODS: We used polymerase chain reaction (PCR) to investigate the prevalence of LMP-1 gene variants in EBV positive lymph nodes from 60 HIV-negative Italian patients with benign and malignant lymphoid disorders. RESULTS: The 30 bp variant was detected in 10 of 39 (25.6%) malignant lymphomas but also in 4 of 13 (30%) reactive lymphadenitis with follicular hyperplasia. Of note is the fact that the 69 bp variant was detected in three cases of malignant lymphoproliferation but also in two cases of localized Castleman's disease of hyalin vascular type. CONCLUSIONS: The molecular detection of the oncogenic variants of the LMP-1 gene in a lymph node biopsy as an indicator of the aggressiveness of the EBV-associated lymphoproliferative disease must be considered with caution. The relatively high frequency of the 69 bp variant in our series compared with that reported in the literature probably reflects a different incidence of LMP-1 variants in healthy populations from different geographical areas. PMID- 10370792 TI - Does a gene in the Xq28 region increase the risk of non-Hodgkin's lymphomas? Working Group for the Epidemiology of Hematolymphopoietic Malignancies in Italy. PMID- 10370793 TI - Paclitaxel and gemcitabine in advanced non-nasopharyngeal head and neck cancer: a phase II study conducted by the Hellenic Cooperative Oncology Group. AB - BACKGROUND: Paclitaxel as monotherapy or in combination with other drugs has demonstrated significant activity in patients with squamous cell carcinoma of the head and neck region (SCCHN). Preclinical studies have shown gemcitabine to be highly active in SCCHN cell lines. PURPOSE OF THE STUDY: To evaluate the activity and toxicity of the combination of paclitaxel by three-hour infusion and gemcitabine as first-line chemotherapy in patients with recurrent and/or metastatic head and neck cancer (HNC). PATIENTS AND METHODS: From September 1996 until May 1998, 44 patients with non-nasopharyngeal recurrent and/or metastatic HNC entered the study. There were 37 men and seven women with a median age of 61 years (range 35-79) and a median performance status of 1 (range 0-2). The location of the primary tumor in the majority of them was either the larynx or the oral cavity. Treatment consisted of six cycles of gemcitabine 1100 mg/m2 over 30 min on days 1 and 8 immediately followed on day 1 by paclitaxel 200 mg/m2 by three-hour infusion. The treatment was repeated every three weeks. RESULTS: Twenty-four (55%) patients completed all six cycles of treatment. A total of 205 cycles were administered, 165 (81%) of them at full dose. The median relative dose intensity (DI) of gemcitabine was 0.93 and of paclitaxel 0.95. Except for alopecia, which was universal, grade 3-4 toxicities included neutropenia (21%), thrombocytopenia (5%), anemia (5%), infection (5%), flu-like syndrome (5%) and peripheral neuropathy (2%). Five (11%) patients achieved complete and 13 (30%) partial responses, for an overall response rate of 41%. After a median follow-up of 13 months, the median time to progression was four months and median survival nine months. CONCLUSIONS: The combination of paclitaxel and gemcitabine is active and well tolerated in patients with recurrent and/or metastatic HNC-randomized studies comparing this combination with other regimens are warranted. PMID- 10370794 TI - Repetitive high-dose therapy with ifosfamide, thiotepa and paclitaxel with peripheral blood progenitor cell and filgrastim support for metastatic and locally advanced breast cancer: results of a phase I study. AB - BACKGROUND: This phase I study was designed to determine the optimal dosages of a novel repetitive high-dose therapy regimen for patients with metastatic breast cancer (MBC). PATIENTS AND METHODS: The planned treatment was three cycles of high-dose ifosfamide, thiotepa and conventional-dose paclitaxel delivered every 28 days with progressive dose-escalation in successive cohorts. Each cycle was supported by peripheral blood progenitor cells (PBPC) and filgrastim. RESULTS: Twenty-three patients were entered into this trial. Of the planned 69 treatment cycles, 59 were delivered and fifteen patients completed all three cycles. The dose-limiting toxicities were renal tubular acidosis, encephalopathy, mucositis and enterocolitis. There was one treatment-related hemorrhagic death. CONCLUSIONS: The recommended doses for phase II or III studies are ifosfamide (10 g/m2), thiotepa (350 mg/m2) and paclitaxel (175 mg/m2). PMID- 10370795 TI - Watchful monitoring for hepatitis B reactivation after intensive chemotherapy. PMID- 10370797 TI - Legal regulations of tissue and organ transplantation in Poland. PMID- 10370796 TI - The role of adhesion molecules in allotransplanted islet cells rejection. Prolongation of islet cells allograft survival by antiadhesion treatment. AB - Insulin-dependent diabetes mellitus is an autoimmune disease caused by the selective destruction of islet beta cells. Allo or xeno transplantation of islet cells may establish a novel promising method of IDDM therapy. Understanding how lymphocytes recognize beta cell antigens is essential for the elucidation of the pathogenesis of islet dysfunction. Leukocyte adhesion to the target cells (endothelium, islets) via adhesion molecule pathways plays an important role in auto and allo/xeno antigen recognition and effector cytodestruction of target cells. However, the expression of these molecules on the endothelium and islet cells during the rejection process still remains unclear. There are some publications describing possible roles of these antigens in the response to the graft. The expression of some of adhesion molecules may contribute to a new method for the diagnosis of graft rejection and its therapy when adhesion blocking substances are used for the treatment. PMID- 10370798 TI - Pathways of antigen traffic from skin allotransplant to recipient lymph nodes--an evolutionarily developed route for initiation of rejection of foreign antigens. AB - Immune events developing in the bed of skin allograft and draining lymphatics and lymph nodes are probably a copy of what happens in skin after invasion by bacteria, viruses or fungi. The mechanism of local immune response developed in skin during the evolution and is highly conserved and efficient in elimination of foreign antigens. This is why the take of a skin allograft is so difficult and immunosuppressive measures applied after allogeneic skin transplantation remain so ineffective. Authors present their results of studies on the human skin immune humoral and cellular factors, underlining their specificities and differences compared with blood. They also analyze the role of non-immunological factors, such as tissue fluid formation and lymph flow in transportation of alloantigen to the lymph nodes. The migratory properties of immune cells are an indispensable factor in transfer of alloantigen to the lymph nodes. Understanding of the evolutionarily developed immune events in skin may allow to analyze the process of skin allograft rejection and can give hints for more effective immunosuppressive policy. PMID- 10370799 TI - Monitoring of rat islet allografts with dithizone after induction of donor specific transplant tolerance by intrathymic administration of soluble alloantigens. AB - Transplantation of whole pancreas or pancreatic islets remains a promising approach to treatment of diabetes mellitus. Since there is no efficient method presently known for in vivo detection of pancreatic islet rejection, we have utilized dithizone [DTZ] to monitor the survival of transplanted islet allografts following the induction of tolerance by a new strategy of deliberate introduction of donor antigens into the adult thymus. In this study, we examined the morphology of islet allografts in vivo and in vitro following pretreatment with intrathymic (IT) inoculation of 2 mg soluble Ag obtained from 3M KCl extracts of resting T-cells with or without ALS immunosuppression in the WF-to-Lewis combination. Fresh isolated rat islets stained pink 3-5 minutes following exposure to medium containing 0.12 mM DTZ solution in DMSO. Intravenous (i.v.) injection of DTZ solution into unmodified recipients of islet allografts that had rejected their grafts showed massive degranulation of islets which did not stain pink with DTZ. This was confirmed by microscopic finding of fibrosis and lymphocytic infiltration. In contrast, i.v. injection of DTZ solution into long term recipients of islet allografts at 50, 100, and 150 days after transplantation showed viable islet cells which stained crimson red with DTZ and the findings were confirmed with microscopic sections. This study demonstrates that DTZ is an effective means of in vivo and in vitro identification of transplanted pancreatic islets and suggests that this strategy may have potential clinical application in the diagnosis of the pancreatic islet rejection. PMID- 10370800 TI - Sinusoidal lymphocytes of liver graft may limit the recurrence of hepatic tumor in liver transplant recipients. AB - The liver immune function is associated with specific lymphocyte population transiently marginated in the liver sinusoids. These cells are of blood origin, however they are phenotypically and functionally different from peripheral blood lymphocytes. The question arises whether tumor proliferating in the liver can modify cell recruitment and function of marginating sinusoidal lymphocytes. Studies were performed in Wistar rats. Livers of normal and colon cancer (induced by i.v. injection of CC531 cells) metastases bearing rats were perfused for sinusoidal lymphocyte isolation. Our studies showed no difference between the number of lymphocytes retained in sinusoids of tumor bearing and normal rats. T lymphocyte subsets remained in similar proportions in liver with colorectal metastases as in normal rats. Long lasting presence of tumor in the liver was accompanied by decreased cytotoxic activity of liver sinusoidal lymphocytes, whereas it had no influence on cytotoxicity of peripheral blood lymphocytes. Repopulation of tumor liver with peripheral blood cells restored cytotoxic activity of sinusoidal lymphocytes. PMID- 10370801 TI - Correlation between heart rate variability and hypothermic storage in human cardiac transplant recipients. PMID- 10370802 TI - The incidence of malignancy in heart transplant recipients. AB - OBJECTIVES AND METHODS: 219 heart transplant recipients with survival over 3 months were retro- and prospectively analysed for the incidence of primary neoplasms. Patients received immunosuppressive drugs (cyclosporine A, azathioprine, steroids) with a 4-5 days induction course of Rabbit Anti-Thymocyte Immunoglobulin (RATG) or monoclonal antibodies induction /OKT3/ in some cases. Anti-rejection treatment consisted of pulse doses of methyloprednisolon or RATG. RESULTS: 9 cases of malignancy (4.1%) with one case of pre-malignant liver condition (dysplasia gigantocellulare, 0.45%) were found (8M; 1F; age: 45-67 y.o., x57.7). Symptoms of neoplasms occurred 7-79 months (x31.4) postoperatively. Skin carcinomas: planoepitheliale, spinocellulare, soft tissue neoplasms/mesenchymal sarcoma, larynx Ca planoepitheliale, lung: adenocarcinoma and Ca microcellulare, kidney Ca clarocellulare and post transplant non-Hodgkin lymphoma were diagnosed. Chemo- and radiotherapy, surgery and reduction of immunosuppression did not change the outcome of malignancy in 6 pts.; (regression 1 pt was., remission-2 pts). Patients died 7-86 months after Htx (x41), 4-25 mos. (x12.5) after suffering from first symptoms and 0-10 months (x4.9) after pathology-based diagnosis of neoplasm. CONCLUSIONS: Heart transplant recipients have an increased risk of carcinogenesis. The incidence of malignancies in the studied group is similar or even lower than in other reports. PMID- 10370803 TI - [Cytokines and growth delay during chronic inflammatory diseases in children]. PMID- 10370804 TI - [The value of procalcitonin in neonatal infections]. AB - The value of procalcitonin (PCT) measurement is not presently completely assessed for the diagnosis of neonatal infections. PATIENTS AND METHODS: This parameter was assessed in a prospective study in the neonatal intensive care unit of Clermont-Ferrand Hospital (France) in comparison to C-reactive protein. All newborn infants admitted before 24 h of life (day 0) in the neonatal intensive care unit were included in the study. Newborns (102) were assigned to one of four groups: group 1: non-infected newborns (n = 41); group 2: possibly infected newborns (n = 33); group 3: probably infected newborns (n = 10); group 4: confirmed infections (n = 18 bacterial or fungal infections). C-reactive protein and PCT were determined in the sera at D0, D1, D3 and D8. We determined the optimal cutoff value of PCT using the Receiver Operating Characteristic curves (R.O.C.). RESULTS: The cutoff value is 1.5 ng/mL at D0 and 10 ng/mL at D1. PCT cutoff value is significantly higher at D1 because of a significant PCT peak on the first day of life independent of any infectious stimulus. Our study shows that at D0 and D1 infected newborn infants had significantly higher mean PCT and C-reactive protein values than non infected newborn infants. C-reactive protein has a better specificity but PCT has better sensitivity and negative predictive value. CONCLUSION: PCT seems to be an interesting marker of neonatal infections especially during the first 24 h of life even though the mechanism of PCT synthesis remains unclear. PMID- 10370805 TI - [Adverse effects of the vaccines Tetracoq, IPAD/DTCP and DTCP. A French study of regional drug monitoring centers]. AB - On request of the French Drug Agency, the Regional Pharmacovigilance Center (RPVC) of Tours has been in charge of the analysis of adverse events (AEs) associated with tetravalent vaccines IPAD/DTCP, DTCP and Tetracoq, and reported to the RPVC or to the pharmaceutical companies that produce them. METHODS: All AEs spontaneously reported during use of one of these vaccines to one of the French Pharmacovigilance Centers or to the responsible firms between January 1, 1986 and December 31, 1990 were take into account. An AE was noted as "serious" in accordance with the European criteria. The incidence of adverse effects was estimated by evaluating the ratio of adverse effects and the number of sales of the vaccine for the same period. RESULTS: From 1986 to 1990, 631 AEs (with 19 duplicate cases) associated with tetravalent vaccines in 606 children (75% < 1 year) were reported. The most frequent AEs were: local AEs at the site of injection (43%), neurologic disorders (12%), hyperthermia (10%) and allergic reactions (10%). Serious AEs represented 25% of all AEs and were similar to those usually described with these vaccines, particularly persistent crying (23), febrile seizures (12), apyretic seizures (14), uneasiness (28) and, rarely, shock (3). CONCLUSION: Incidences of AEs reported with pentavalent vaccines are very low, probably underestimated because of the under-notification by prescribers of AEs of vaccines licensed some time ago. It will be interesting to compare these data with AEs of penta- and hexavalent vaccines since they have replaced tetravalent vaccines. PMID- 10370806 TI - [Retrospective study of 32 cases of intraosseous perfusion]. AB - BACKGROUND: We studied all intraosseous infusions performed between 1994 and 1997 by the pediatric intensive care unit and by the pre-hospital emergency medical staff in the Hopital d'Enfants, Toulouse, France. POPULATION AND METHODS: We report 32 cases of intraosseous infusions in 30 children aged 2 weeks to 9 years. RESULTS: In our population, such a technique has been used in about 60% of all cardiopulmonary arrest, drowning or traffic accident cases. Intraosseous infusion was successful in all cases, on the first attempt in more than 80% of cases. Nine children recovered without any sequelae. No major complications have been observed. CONCLUSIONS: Intraosseous infusion is safe, rapid and effective. It is an essential alternative route in pediatric resuscitation when no other venous access can be performed quickly. An effort must be made on behalf of its diffusion and teaching. PMID- 10370807 TI - [Growth curves from birth to 6 years of age: growth in weight, height and cranial circumference according to sex]. AB - The growth supervision of children using growth curves is a widespread and useful tool in general pediatric practice. In France the latest reference curves are rather ancient, therefore it seemed to us interesting to re-examine some growth parameters and to compare them to the current reference data. PATIENTS AND METHODS: The studied sample was composed of 7,000 children from the Rhone-Alpes region in France who were seen for a school health check-up. Anthropometric measurements (35,000) related to weight, height and cranial circumference of these children aged from 0 to 6 were selected from their health booklets. Centile curves for these three variables were drawn from these measurements using the LMS method, which is specifically suited to these types of data. RESULTS: If height and cranial circumference can be considered as normally distributed, weight is markedly skewed to the right, reflecting a high prevalence of children with heavy weight. Comparison with current references data shows more or less similar results for weight, height and cranial circumference: the medians and the 2.5 centile are constantly higher in our sample than those of the reference data (the discrepancy increasing with age). The differences are more important regarding the part of the distribution which concerns the highest values: overall, the 90th centile of our distribution corresponds rather closely to the 97.5th one of the current reference data. Thus, at 6 years of age, the 97.5th centile of the reference weight distribution for girls is 23,400 kg, while it is estimated at 27,770 kg in our sample. One should take in account the different characteristics of the two studies to interpret these differences. CONCLUSION: The aim and use of such growth curves is discussed, together with recent computer applications in this field. PMID- 10370808 TI - [An unusual care of intestinal invagination: jejunojejunal invagination]. AB - BACKGROUND: Jejunal intussusception is uncommon in comparison with ileocolic form. It is more frequent in children over 2 years of age and has an atypical subacute presentation. An underlying anatomical cause is usually found. CASE REPORT: A 14-year-old boy was admitted for abdominal pain with bilious vomiting. The physical examination was normal, with only the ultrasonography showing an intussusception in the left hypochondrium. At laparotomy the diagnosis of jejunal intussusception was made; its reduction was impossible. A resection and end to end anastomosis was performed. The anatomopathology examination found a polyp in ectopic gastric mucosa. CONCLUSION: Jejunal intussusception must be better understood as its diagnosis could be made too late. Surgical exploration is the treatment of choice because of the usual underlying anatomical cause. PMID- 10370809 TI - [Neonatal toxic erythema: 3 atypical cases]. AB - Diagnosis of pustular dermatosis occurring during the first days of life is based on clinical findings. Erythema toxicum neonatorum (ETN) is the more frequent benign self limiting eruption in the newborn. CASE REPORTS: Three cases of ETN with localized pustules to the genitals and perineal area are described. COMMENT: When encountering a newborn with a localized pustulosis rash, it is important to separate benign condition as ETN from those that require prompt diagnosis and therapy. Atypical ETN and pustular dermatosis due to bacterial or viral infections or inflammatory diseases (e.g., eosinophilic pustulosis) can be differentiated by cytological and bacterial samples. PMID- 10370810 TI - [Smith-Lemli-Opitz syndrome]. AB - BACKGROUND: Smith-Lemli-Opitz syndrome (SLOS) is an autosomic recessive metabolic affection. Children affected by SLOS exhibit a defect in cholesterol biosynthesis associated with a high concentration of cholesterol precursor 7 dehydrocholesterol (7 DHC) and its isomers, which is due to an enzymatic block at the level of delta-7-DHC reductase. SLOS has been subdivided into two types on the basis of clinical severity: type I is the classic and type II is the severe one. CASE REPORT: A full term female was born from a pregnancy complicated by oligoamniosis and intra-uterine growth retardation. The neurologic status was immediately impaired with severe hypotonia, absence of reflexes, and abnormal crying. She exhibited multiple congenital anomalies with a facial dysmorphia, anomalies of members, unicornus uterus and a pyloric stenosis. Plasmatic concentration exhibited a normal cholesterolemia contrasting with an elevated level of 7 and 8 DHC. Major alimentary tract defect led to enteral and parenteral nutrition. The severe neurological defect led to death on the 16th day of life. CONCLUSION: Despite normal blood cholesterol levels that can be attributed to enteral and parenteral nutrition, the severity of clinical findings and the lethal course permit to classify this case as type II. PMID- 10370811 TI - [Favorable outcome of treatment with NTBC of acute liver insufficiency disclosing hereditary tyrosinemia type I]. AB - BACKGROUND: Hereditary tyrosinemia type I is a disease with a severe prognosis. Main causes of death are acute liver failure, neurologic crises and hepatocarcinoma. NTBC, which acts as an inhibitor of the 4-hydroxyphenylpyruvate dioxygenase, prevents the formation of toxic metabolites involved in hepatic, renal and neurologic lesions. CASE REPORTS: Results of NTBC therapy used in three infants with type I tyrosinemia who presented with acute liver failure are reported. The diagnosis relied on the finding of high plasmatic levels of tyrosine and methionine, and abnormal urinary excretion of succinyl acetone and delta aminolevulinic acid. Treatment with NTBC was initiated within 2 to 8 days from onset of symptoms. Signs of liver failure resolved after 3 weeks therapy. After 12 to 39 months of follow-up, outcome remains favorable. CONCLUSION: The results reported here highlight the efficiency of NTBC in type I tyrosinemia with early acute onset. However, the long term outcome needs to be determined with regards to prevention of hepatocarcinoma and toxicity of the drug. PMID- 10370812 TI - [Fetal endoscopic surgery: between dreams and reality. (South-East medicine and prenatal research)]. AB - "Open" fetal surgery has had little success in the management of fetal malformations, and is limited by two important unresolved difficulties: control of the operating stress on the fetus and maternal tocolysis. The "mini-invasive" approach by endoscopic surgery circumvents these difficulties and appears promising. Decision making with the dilemma between prenatal or postnatal surgery remains critical. PMID- 10370813 TI - [For or against the early use of nasal continuous positive pressure and exogenous surfactant in hyaline membrane disease. Physiopathologic arguments]. AB - The use of nasal CPAP in the treatment of respiratory distress syndrome in very premature newborns follows pathophysiological basis. The authors emphasize the usefulness of nasal CPAP and surfactant in the treatment of respiratory distress syndrome. The aim of this strategy is to reduce alveolar atelectasis, thus reducing the incidence and the severity of respiratory distress syndrome, together with a possible reduction of the incidence of bronchopulmonary dysplasia. PMID- 10370814 TI - [Antiretroviral therapy and prevention of maternal-fetal transmission of HIV-1. Current and future strategies]. AB - Azidothymidine is effective and recommended for the prophylaxis of vertical HIV transmission. Data regarding this treatment have been collected over the last decade, leading to it being widely prescribed despite the lack of information concerning its long term toxicity. Antiretroviral drug combinations administered during pregnancy appear to ensure a better protection of both mothers and their offspring. However, data available on the adverse effects of these therapies during pregnancy are scarce and mainly obtained from in vitro or animal models. Therefore there is a need for multicentric trial including long-term follow-up of exposed patients. PMID- 10370815 TI - [Radiologic ase of the month. Double aortic arch detected during esophageal pH metry]. PMID- 10370816 TI - [Attitude of pediatricians toward adopted children from foreign countries]. AB - Adopted children from foreign countries represent a high risk population for infectious diseases, nutritional problems and neuro-developmental delay. Medical screening including clinical and biological evaluation is recommended after arrival. PMID- 10370817 TI - [Post-traumatic stress syndrome in children]. AB - Post traumatic stress disorder occurs in children as well as in adults following a stressful traumatic event, either unique and exceptionally severe, or recurrent as in abused children. The main symptoms are repetition, avoidance and neurovegetative activation. Prevention and recognition of this disorder are important. Parents must be sensitized to the necessity of an early management. PMID- 10370818 TI - [Analgesia using a (50/50) mixture of nitrous oxide/oxygen in children]. AB - Nitrous oxide is a gas that has been used to provide analgesia to patients for more than a century. Its modern use started in the late sixties when a mixture of 50% nitrous oxide/50% oxygen was prepared. Nitrous oxide/oxygen provides analgesia within 3 minutes of inhalation and this analgesic effect disappears in less than 4 minutes after cessation. Its administration is very easy and a complete or partial pain relief is observed in 75 to 81% of patients. The gas mixture has been found to be safe with few side effects and no significant adverse reactions. Diffusion hypoxia which could lead to hypoxemia was reported in 1955, but recent work does not confirm this complication. Nitrous oxide/oxygen mixture constitutes an excellent drug to control procedure-related pain in children. This articles describes the history, the pharmacology, and the clinical uses of nitrous oxide/oxygen in children. PMID- 10370819 TI - [The usefulness of statistical tests in publications]. PMID- 10370820 TI - [Precocious puberty in adopted children, a risk not to be forgotten]. PMID- 10370821 TI - [Streptococcus group A adenitis and bacteremia complicating varicella]. PMID- 10370822 TI - [Isolated and persistent elevation and of transaminases (aspartate aminotransferase) using a macroenzyme]. PMID- 10370823 TI - [Pediatrics and family politics]. PMID- 10370824 TI - [The prevention of anemia in premature infants: the role of recombinant human erythropoietin in a level III neonatology department]. PMID- 10370825 TI - [In children, social determinants of health are changing]. PMID- 10370826 TI - Alternatives to allogeneic blood transfusions. PMID- 10370827 TI - Complications of regional blocks: what do we really know? PMID- 10370829 TI - Management of the severely head injured patient. AB - The care of the severely brain injured patient presents important challenges to the anesthesiologist. The patients are often young, there is little time for preparation, there are associated injuries, the airway management may be difficult or risky to the brain or spinal cord, the outcome varies widely and it is not always possible to monitor the desired physiological variables. Furthermore, there is no single intervention or agent that has emerged as being beneficial, but the degree to which care is taken in the domain of the anesthesiologist, will have an important impact on outcome. The central themes in patient care include rapid attention to resuscitation and maintenance of adequate CPP and oxygen delivery. Careful attention to post-operative care will also affect outcome. Recognizing that the outcome may not be determined solely at the time of impact is an important concept for all who take care of these patients. PMID- 10370828 TI - Adverse outcomes in ambulatory anesthesia. PMID- 10370830 TI - The prevention and treatment of cerebral ischemia. AB - Although the major focus of recent cerebral protection research has been aimed at developing receptor-specific drugs, this effort has currently resulted in few improvements in patient outcome. Until advances in pharmacology translate to improvements in humans, the clinician and his patients will be well served by using more traditional techniques to prevent and treat cerebral ischemic events. This approach will involve interventions to a) identify patients who are experiencing or are at risk for developing cerebral ischemia, and b) alter systemic physiology in an attempt to lessen the duration and severity of any ischemic insults. Initial therapy should include interventions to improve cerebral perfusion and the oxygen carrying capacity of the blood. Once this is accomplished, measures should be taken to control blood glucose concentrations and treat fever. In otherwise stable surgical patients, mild reductions in patient temperature also may be of benefit, provided the temperature reductions do not introduce problems in systemic physiology and the patient is rewarmed prior to awakening from general anesthesia. General anesthetic choice may be of importance in controlling intracranial pressure and seizure activity; however, if direct cerebral protection is desired, the anesthetic of choice should be a barbiturate. Finally, in the patient at risk for cerebral vasospasm, nimodipine treatment should be considered. Collectively, these interventions should increase the patient's chance for optimal neurologic recovery following ischemia. PMID- 10370832 TI - Perioperative factors influencing surgical morbidity: what the anesthesiologist needs to know. PMID- 10370831 TI - Death in the ICU--the Halifax experience. PMID- 10370833 TI - Awareness during anesthesia. PMID- 10370834 TI - The new relaxants: are they worth it? PMID- 10370835 TI - Anesthesia for minimally invasive surgery. PMID- 10370836 TI - Management of controversies in obstetric anesthesia. PMID- 10370837 TI - Practical pharmacokinetics as applied to our daily anesthesia practice. PMID- 10370838 TI - Risks and benefits of the practice of anesthesiology. PMID- 10370839 TI - Anemia, hypoxia and hypercapnia thresholds. Lessons from physiological limits in critically ill patients. AB - Physiological alterations occur in the critical care medicine and reflect illness. Rendering patients physiologic parameters in the range that is normal for the population is not necessarily good; it may be frankly harmful. We do not currently possess outcome-based tools that allow us to titrate physiological parameters and ensure improved outcome. It is highly unlikely that our practice will evolve to inducing anemia, hyperthermia, hypoxemia, hypercapnia and hypotension in our critically ill patients! However, evolving knowledge of the appropriate thresholds for these parameters in critically ill patients, in addition to greater understanding of the potential iatrogenic illness associated with parameter 'normalization', could lead to provision of enhanced patient care. In the coming years it is possible that we will redefine the 'normal' range for common clinical and laboratory values relating to the critically ill. We may switch to 'context sensitive' interpretation of parameters of illness, and manage critically ill patients accordingly. PMID- 10370840 TI - Current evaluation and management of vulvovaginitis. AB - There are many problems in the diagnosis and treatment of vaginitis. Often, the patient is not examined (telephone treatment) or examined improperly with lack of attention to the wet prep. In patients with recurrent vaginitis, it should not be assumed that the current infection is the same as a previous infection without a thorough examination. At times, there is an overuse of topical steroids for all vulvar symptoms or use of antifungals for all vulvar symptoms. The various abnormalities in vulvovaginitis have unique physical findings, laboratory tests, and treatments. It should be remembered that unusual conditions of the vagina and vulva may resemble vulvovaginitis. Many vulvar conditions must be considered when a patient reports discharge and itching. It is important to remember that if the treatment is not working, reconsider the diagnosis. PMID- 10370841 TI - Ambulatory management of uterine leiomyomata. PMID- 10370842 TI - Advances in the diagnosis and treatment of human papillomavirus infections. AB - During the past decade, much has been learned about the natural history of HPV. Most infections occur early in one's sexual life. The overwhelming majority of infections are cleared by the host immune system and never present as warts or neoplasia. Certain patient behavior such as smoking, frequent sex with many different partners, other STD infections, especially HIV, and immune-suppressive drugs promote HPV expression and cause persistent infection. Persistent HPV infection is very strongly related to neoplasia. In addition to high-risk HPV types, variant subtypes have been identified that interact with the host immune system to subvert host immunity and encourage viral persistence. New treatment programs rely on drugs that modulate the immune system and disrupt viral persistence. There is a real possibility of HPV vaccines in the future, which may protect the unexposed patient. PMID- 10370843 TI - Clinical management of vulvodynia. AB - Vulvodynia represents a group of vulvar disorders that can be clinically perplexing. Unfortunately, there are no simple tests for its diagnosis. Patients often are reluctant to report vulvar pain. Some are embarrassed to reveal their perceived "sexual dysfunction" caused by dyspareunia. Others have been told in the past "It's all in you head," or "It's just a yeast infection." Most patients with vulvodynia are seen by multiple practitioners and are placed on a variety of treatments before a correct diagnosis is made. All too often, these treatments are not only ineffective but also damaging. Careful clinical investigation is required for the correct diagnosis and treatment of vulvodynia (Figure 2). PMID- 10370845 TI - Office assessment of female urinary incontinence. PMID- 10370844 TI - Office management of benign breast disease. PMID- 10370846 TI - Office screening for gynecologic cancer. PMID- 10370847 TI - Office hysteroscopy. AB - Office hysteroscopy has developed into an easy, safe, quick, and effective method of intrauterine evaluation that provides immediate results, offers the capacity of direct targeted biopsies of suspicious focal lesions, and offers the direct treatment of some intrauterine conditions. It has been facilitated by the availability of small-caliber endoscopes. Because of its simplicity and ease, the procedure is applicable as a screening method for patients with abnormal uterine bleeding or questionable hysterograms and for patients with suspected intrauterine pathology. Office hysteroscopy can be undertaken in a short period of time with minimal morbidity and inconvenience to the patient. It is important, nonetheless, to select the patients appropriately and time the examination strictly to the early follicular phase, once menstruation has ceased. When suction aspiration plastic cannulas are used for endometrial sampling, the combined procedure, hysteroscopy-suction sampling, offers an excellent method in the evaluation of patients with abnormal uterine bleeding. Transvaginal sonography with or without fluid enhancement complements the uterine evaluation, rather than replacing hysteroscopy, by outlining intramural uterine lesions such as myomas, adenomyosis, and other adnexal pathology not susceptible to hysteroscopic evaluation. Although some patients may not require analgesia or anesthesia for office hysteroscopy, the majority will benefit from a paracervical block or topical anesthesia, particularly if a suction endometrial aspiration will follow hysteroscopy or if any hysteroscopic intervention is performed, including a targeted biopsy. The success office hysteroscopy depends on the appropriate selection of the patient, the absence of contraindications, adequate instrumentation, and meticulous technique. PMID- 10370848 TI - Office management of early induced abortion. AB - Fertility control, including induced abortion, is an integral part of women's health care. With the availability of sensitive home pregnancy tests, increasing numbers of women are seeking abortion during early pregnancy. Recent technological advances allow women with early pregnancies to choose medical or surgical abortion options. With appropriate skills and preparation, clinicians can offer the full range of early abortion techniques safely in the office setting. Helping women choose the most appropriate method enhances safety, efficacy, and patient satisfaction. PMID- 10370849 TI - Primary care for women: the role of the obstetrician-gynecologist. AB - It is critical to understand that "one size does not fit all" in health care. Each patient is unique and involved in her own destiny. Our goal should be to maximize her health status, to provide health education, and to advocate for her health and well-being. We must implement a spectrum of primary care in the office to further expand the excellent health care that obstetrician-gynecologists have always provided to women. PMID- 10370850 TI - Review of autoimmune thrombocytopenia: pathogenesis, diagnosis, and management in pregnancy. PMID- 10370851 TI - Gestational thrombocytopenia. PMID- 10370853 TI - Overview of platelet physiology and laboratory evaluation of platelet function. AB - Appropriate laboratory testing for the platelet-type bleeding disorders hinges on an adequate assessment in the history and physical examination. Patients with histories and screening laboratory results consistent with coagulation disorders (hemophilia, disseminated intravascular coagulation) are not appropriate candidates for platelet function testing. In contrast, patients with a lifelong history of platelet-type bleeding symptoms and perhaps a positive family history of bleeding would be appropriate for testing. Figure 6 depicts one strategy to evaluate these patients. Platelet morphology can easily be evaluated to screen for two uncommon qualitative platelet disorders: Bernard-Soulier syndrome (associated with giant platelets) and gray platelet syndrome, a subtype of storage pool disorder in which platelet granulation is morphologically abnormal by light microscopy. If the bleeding disorder occurred later in life (no bleeding with surgery or trauma early in life), the focus should be on acquired disorders of platelet function. For those patients thought to have an inherited disorder, testing for vWD should be done initially because approximately 1% of the population has vWD. The complete vWD panel (factor VIII coagulant activity, vWf antigen, ristocetin cofactor activity) should be performed because many patients will have abnormalities of only one particular panel component. Patients diagnosed with vWD should be classified using multimeric analysis to identify the type 1 vWD patients likely to respond to DDAVP. If vWD studies are normal, platelet aggregation testing should be performed, ensuring that no antiplatelet medications have been ingested at least 1 week before testing. If platelet aggregation tests are normal and if suspicion for an inherited disorder remains high, vWD testing should be repeated. The evaluation of thrombocytopenia may require bone marrow examination to exclude primary hematologic disorders. If future studies with thrombopoietin assays confirm preliminary results, however, the bone marrow examination of certain patients may be replaced by a thrombopoietin level. PMID- 10370852 TI - Alloimmune thrombocytopenia. AB - For the fetuses who are at risk for antenatal or postnatal sequelae from AIT, prevention and treatment are now possible. This requires the attention of the obstetrician to factors in the patient's history and early referral to a center experienced in the diagnosis and management of fetal AIT. PMID- 10370854 TI - Diagnosis and management of thrombotic microangiopathies during pregnancy. PMID- 10370855 TI - Clinical uses of intravenous immunoglobulin in pregnancy. AB - The obstetric literature contains numerous reports of IVIG therapy for various conditions encountered during pregnancy. The mechanisms of action of IVIG are uncertain and may vary depending on the specific disorder. Immunoglobulin G infusions appear to be well tolerated by the parturient. The occurrence of major and minor side effects is uncommon, and infectious morbidity is low. Further research will be necessary to elucidate the specific mechanisms of action of IVIG in certain disease states. Determining the exact "therapeutic agent" in IVIG for each specific disease state may allow for a more tailored approach to treatment (i.e., isolation or production of the particular antibody). Outcome assessment, long-term positive and negative effects, cost-benefit analysis, and effects on fetal and neonatal immune function require further study through randomized trials. PMID- 10370856 TI - HELLP syndrome. PMID- 10370857 TI - Congenital disorders of platelet function. PMID- 10370858 TI - James Cowles Prichard's anthropology: remaking the science of man in early nineteenth-century Britain. PMID- 10370859 TI - Behavior and pituitary-adrenal function in large white and Meishan pigs. AB - Six-wk-old piglets of both sexes from European Large White (LW, n = 36) and Chinese Meishan (MS, n = 24) breeds were individually exposed to a novel environment, a stressful stimulation. Behavioral and pituitary-adrenal reactivity were investigated. When compared with LW, MS pigs displayed low locomotion (18.5 +/- 2.2 vs. 41.0 +/- 3.8 squares crossed/10 min; P < 0.0001), and defecation scores (0.58 +/- 0.15 vs. 4.86 +/- 0.37 fecal boli; P < 0.0001). Basal concentrations of cortisol were higher in MS (96.1 +/- 1.1 vs. 44.9 +/- 1.1 ng/ml; P < 0.0001), although no differences between breeds were found in basal concentrations of adrenocorticotropic hormone (ACTH). In response to novel environment exposure, the ACTH increase was greater in LW than in MS, but the cortisol response was not different on a log scale. To further investigate the pituitary-adrenal differences between the two breeds, the 24-hr profile of ACTH and cortisol plasma concentrations, a corticotropin-releasing factor (CRF) and a coupled dexamethasone-ACTH test were studied. Five castrated male 9-wk-old piglets from each breed were fitted surgically with a jugular vein catheter. A classic marked circadian rhythm of cortisol and a weak nycthemeral variation of ACTH were found. Cortisol concentrations were approximately twice higher in MS exclusively during the early light phase (from 0800-1200 hr) of the cycle, but no significant interbreed difference was found in the circadian rhythm of ACTH. Administration of CRF (1 microgram/kg iv) induced the same significant increase in plasma ACTH and cortisol concentrations in both breeds. Administration of ACTH (10 micrograms/kg i.v.) increased significantly cortisol concentrations and revealed no difference in plasma cortisol response to ACTH. These data suggest that the hypercortisolism of MS pigs is of adrenal origin, and related to extrapituitary factors that control the adrenal sensitivity during the light phase of the diurnal cycle. PMID- 10370860 TI - Effect of immunization against melatonin on prolactin concentrations and the timing of reproductive transitions in ewes. AB - The objective of this experiment was to develop a procedure for immunizing ewes against melatonin that would alter the effects of changing photoperiod on seasonal reproduction and prolactin secretion. Ewes were immunized against human serum albumin (HSA) as controls (n = 9) or a melatonin-human serum albumin conjugate (0.25 mg; n = 10) on December 14th (Day 0) and boosted 9 times. They were maintained on natural photoperiod and then transferred indoors and exposed to long days for 35 d, followed by short days for 146 d, long days for 93 d, and short days for a further 123 d. Antibody titers to melatonin (at a serum dilution of 1:1,250) were significantly higher in immunized ewes (27.3 +/- 6.6%) than controls (0.7 +/- 0.1%; P < 0.001). At the end of the experiment, antibody titers in immunized ewes (at dilution of 1:50) were higher in blood (43.7 +/- 8.2%) than in cerebrospinal fluid (10.8 +/- 3.9%; P < 0.05), and highly correlated (r2 = 0.746). Onset of the breeding season was advanced slightly after the second transfer from long to short days in immunized ewes (April 12 +/- 3 d) compared with controls (April 25 +/- 3 d; P < 0.05). Mean serum prolactin concentrations were lower (P < 0.05) in melatonin-immunized ewes compared with controls on natural photoperiod, after transfer from long to short days, during long days, and after the second transfer from long to short days. In conclusion, despite melatonin-immunization increasing antibody titers in blood and cerebrospinal fluid, and decreasing prolactin concentrations over much of the experiment, minimal effects on the timing of reproductive transitions in the ewes were evident. This discrepancy between the response of the prolactin and reproductive axes to melatonin immunization supports the hypothesis of a dual site of action of melatonin, with melatonin acting in the pituitary gland to mediate the effects of photoperiod on prolactin secretion and in the mediobasal hypothalamus to affect reproductive responses. PMID- 10370861 TI - Action of long(R3)-insulin-like growth factor-1 on protein metabolism in beef heifers. AB - Insulin-like growth factor-1 (IGF-1) is perhaps the most important endogenous factor controlling growth. Most studies to date in livestock have shown that IGF 1 has greatest efficacy when animals are in a catabolic state. We have determined the effects of an i.v. infusion of the IGF-1 analog Long(R3)-IGF-1 on protein metabolism in beef heifers that were slowly losing liveweight because of restricted feeding. There was a tendency for both whole-body protein and skeletal muscle protein to be conserved in Long(R3)-IGF-1-treated heifers. Long(R3)-IGF-1 administration markedly reduced the plasma concentrations of all amino acids measured and glucose. There was a significant change in the profile differences of endogenous plasma IGF-1 concentrations during the 8-hr infusion period, with plasma IGF-1 decreasing sharply in the test group. There was a significant difference in mean profiles for plasma IGF-2 between the test and control groups. Overall, plasma IGF-2 for the control group decreased only slightly over time (about 40 ng/ml), whereas the test group decreased dramatically (by about 140 ng/ml). Increased plasma concentrations of a 31-32-kDa IGF-binding protein (possibly IGF-binding protein-1) in the treated group was detected by radioligand blot. We found that Long(R3)-IGF-1 infusion tended to preserve whole-body and muscle protein in beef heifers on a low-quality diet, and suggest that further investigation of this treatment may provide an alternative approach to reducing weight loss during the dry season. PMID- 10370863 TI - Relative insensitivity of avian skeletal muscle glycogen to nutritive status. AB - Previous studies in avian species have reported time-dependent losses in muscle glycogen with prolonged feed withdrawal (FW). However, cervical dislocation was used to collect tissues, a method that results in significant involuntary muscle convulsions. In this study, cervical dislocation alone was found to reduce muscle glycogen by 23%, therefore, barbiturate overdose was used to collect tissue samples before and after FW, at the end of refeeding, and from continuously fed controls at each interval. Additionally, plasma samples from 6-wk-old male chickens were taken at the initiation and end of a 24-hr feed withdrawal, and at various times during refeeding. After 24 hr of FW, liver glycogen decreased markedly (77%; P < 0.05), whereas muscle glycogen decreased slightly and transiently, such that it returned to and remained at control levels, even after prolonged (72 hr) FW. Plasma glucose was decreased, whereas glucagon was elevated after a 24-hr feed withdrawal (P < 0.05), when compared with control concentrations. Muscle glycogen levels were not significantly increased over control levels after refeeding, but liver glycogen was increased by 380% (P < 0.05). Feed deprivation followed by refeeding resulted in increased circulating insulin and glucose levels when compared with control levels. Therefore, by using methods of tissue collection that ensure that muscle glycogen determinations are not confounded by artifactual degradation, these results verify that regulation of avian muscle glycogen stores is similar to that in mammals. PMID- 10370862 TI - The drop in plasma thyrotropin concentrations in fasted chickens is caused by an action at the level of the hypothalamus: role of corticosterone. AB - Fasting has severe effects on thyroid metabolism in the chicken: plasma thyroxine (T4) concentrations increase, whereas 3',5,3-triiodothyronine (T3) concentrations decrease. In the present report we studied the effect of fasting at the level of: 1) the pituitary (plasma thyrotropin (TSH) concentrations; the sensitivity of thyrotrophs to corticotropin-releasing hormone (CRH) and TSH-releasing hormone (TRH)); and 2) the hypothalamus (TRH content). A regulatory role of corticosterone is discussed. One day of fasting resulted in a drop in plasma TSH concentrations. Fed and nonfed animals were treated with ovine CRH (oCRH) or TRH. The sensitivity of thyrotrophs to the respective hypothalamic hormones was increased when animals were subjected to a 1-d period of fasting. A 75% (TRH) and 50% (oCRH) increase in plasma TSH was recorded in fasted animals, whereas both secretagogues did not evoke any response in their fed counterparts. The drop in plasma TSH cannot, therefore, be attributed to a loss in sensitivity of thyrotrophs to hypothalamic stimulatory control. In an identical experiment, plasma TSH concentrations decreased, whereas hypothalamic TRH content was higher in fasted animals, suggesting a decreased hypothalamic TRH release toward the pituitary. In both fasting experiments, plasma corticosterone concentrations were increased after 1 d of fasting. Because an i.v. injection of corticosterone elevated hypothalamic TRH contents and decreased plasma TSH concentrations, a corticosterone-induced TSH decrease during fasting is suggested through an action at the level of the hypothalamus. PMID- 10370864 TI - Recent progress in research on wine and its components and their favorable effects on health. PMID- 10370865 TI - Advances in the study of secondary metabolites occurring in grapes and wines. AB - In recent years significant advances have been made in the field of secondary metabolites belonging to the polyphenol group and precursors to varietal aromas. Following research on anthocyanins, flavonoids, flavans and phenolic acids of the benzoic and cinnamic type, hydroxystilbenes were thoroughly investigated because of their pharmacological importance. Their presence in the components of grape skins was first noted in 1980. Varietal aromas have mostly been found in their glycoside form. They are known to belong to the class of terpene alcohols, norisoprenoids and benzenoids, though their role in human metabolism is as yet little known. PMID- 10370866 TI - Stilbene compounds: from the grapevine to wine. AB - Stilbenes are natural compounds occurring in a number of plant families, including Vitaceae and (within this family) Vitis vinifera L., which is the most important species grown worldwide for grape and wine production. Stilbenes (resveratrol and viniferins) are present in grapevine as constitutive compounds of the woody organs (roots, canes, stems) and as induced substances (in leaves and fruit) acting as phytoalexins in the mechanisms of grape resistance against certain pathogens. Resveratrol (3, 5, 4'-trihydroxystilbene) was also detected in wine and it was thought to be the active principle of red wines that were shown to reduce heart diseases. This paper reviews data, obtained by the Viticulture Institute of the Catholic University at Piacenza and taken from the literature, on some aspects of stilbene physiology in grapevine and on their relation to resveratrol wine levels. Constitutive stilbene contents of woody organs are reported, as well as the possible role of cluster stems as a source of resveratrol for wine. The accumulation of stilbenes in grape berries infected by grey mould (Botrytis cinerea Pers.) has been investigated and the effects of environmental factors on resveratrol grape and wine levels will be discussed. An unidentified new hydroxystilbene was detected in wine. PMID- 10370867 TI - Cancer chemopreventive activity of resveratrol. AB - Resveratrol (3,5,4'-trihydroxy-trans-stilbene) is a naturally occurring compound shown to inhibit carcinogen-induced preneoplastic lesion formation in mouse mammary organ culture and tumorigenesis in the two-stage mouse skin model. Cancer chemopreventive potential was also suggested in various assays reflective of the three major stages of carcinogenesis. Anti-initiation activity was indicated by its antioxidant and antimutagenic effects, inhibition of the hydroperoxidase function of cyclooxygenase (COX), and induction of phase II drug-metabolizing enzymes. Antipromotion activity was indicated by antiinflammatory effects, inhibition of production of arachidonic acid metabolites catalyzed by either COX 1 or COX-2, and chemical carcinogen-induced neoplastic transformation of mouse embryo fibroblasts. Antiprogression activity was demonstrated by its ability to induce human promyelocytic leukemia (HL-60) cell differentiation. Moreover, pretreatment of mouse skin with resveratrol significantly counteracted 12-O tetradecanoylphorbol-13-acetate (TPA)-induced oxidative stress, as evidenced by numerous biochemical responses. Resveratrol reduced the generation of hydrogen peroxide, and normalized levels of myeloperoxidase and oxidized-glutathione reductase activities. It also restored glutathione levels and superoxide dismutase activity. As judged by the reverse transcriptase-polymerase chain reaction, resveratrol selectively inhibited TPA-induced expression of c-fos and transforming growth factor-beta 1 (TGF-beta 1), but did not affect other TPA induced gene products including COX-1, COX-2, c-myc, c-jun, and tumor necrosis factor-alpha. These data indicate that resveratrol may interfere with reactive oxidant pathways and/or modulate the expression of c-fos and TGF-beta 1 to inhibit tumorigenesis in mouse skin. As reported herein, in addition to the activities described above, resveratrol inhibited the de novo formation of inducible nitric oxide synthase (iNOS) in mouse macrophages stimulated with lipopolysaccharide. This finding suggests an additional mechanism by which resveratrol may function as a cancer chemopreventive agent. PMID- 10370868 TI - Effect of resveratrol and some other natural compounds on tyrosine kinase activity and on cytolysis. AB - Resveratrol is a phytoalexin with several biological and pharmacological activities including the "French paradox". We investigated the effect of resveratrol on cytolytic activity by oxygen reactive species and on soluble and particulate tyrosine kinases from human placenta and human prostatic adenoma. These effects were compared with those of piceatannol, quercetin, catechin and epicatechin. Fifty percent of erythrocyte lysis due to H2O2-lactoperoxidase-KI incubation, in which I3-, OI- and oxygen singlet are produced, was obtained after 22 +/- 7 (SD) min in the absence of the tested compounds. The 50% lysis was obtained after 66 +/- 15, 129 +/- 35, 196 +/- 21, 240 +/- 63 and 420 +/- 80 min with 40 microM piceatannol, quercetin, resveratrol, epicatechin and catechin respectively. Protection was concentration dependent. The assay of tyrosine kinase activity was performed using two different substrates as follows: substrate A corresponded to the sequence 1-17 of gastrin, and substrate B to sequence 6-20 of cell division kinase p34cdc2. In all experiments, initial velocity was measured. When assayed with both substrates, tyrosine kinase activities from particulate and cytosolic fractions of placenta were more inhibited by piceatannol and quercetin. Resveratrol significantly inhibited the particulate fraction and the cytosolic fraction respectively when substrates A and B were employed: Catechin acted as an inhibitor with substrate A and particulate fraction while in the other experimental conditions it acted as an activator. Resveratrol inhibited the tyrosine kinase of particulate and cytosolic fractions of prostatic adenoma assayed with substrate A and B. PMID- 10370869 TI - Effects of resveratrol on the rat brain respiratory chain. AB - The aim of this work was to investigate the possible effects of resveratrol on the mitochondrial respiratory chain in rat brains. Isolation of mitochondria was performed at 4 degrees C using differential centrifugation. Mitochondrial respiration rate (0.4 mg of protein/ml) was determined by measuring mitochondrial oxygen consumption with a Clark electrode at 37 degrees C. Respiratory control ratio (RCR) was evaluated as the state 3/state 4 ratio of oxidative phosphorylation with substrates adenosine 5'-diphosphate (ADP) and malate plus glutamate, respectively in the presence and in the absence of resveratrol. The rate of oxygen consumption by the different complexes was checked using rotenone (2 microM), malonate (10 mM), antimycin A (1 microM), potassium cyanide (KCN) (0.3 mM) and oligomycin (10 microM) to inhibit complexes II, III, IV, V and I, respectively. Moreover, enzyme activity determinations were checked as follows: the activities of complexes II-III were measured as the rate of cytochrome c reduction at 550 nm (37 degrees C) successively triggered either by succinate (complexes II and III) or by decylubiquinol (DUQH2) (complex III), in the presence and in the absence of resveratrol. Adenosine 5'-triphosphate (ATP) synthase activity was checked as ATP hydrolysis (ATPase) at 37 degrees C for 10 min from purified mitochondria on Percoll gradient. The inorganic phosphate (Pi) concentration was measured by the Fiske and Subbarow method. When complexes I to V were activated by glutamate plus malate, resveratrol (10(-11) - 10(-4) M) significantly decreased RC (p < 0.001) following a biphasic curve with two EC50 values, 0.162 +/- 0.072 microM and 24.5 +/- 4.0 microM, representing about 56% of total oxygen consumption inhibition. We also observed a concentration-dependent effect on state 3 with two EC50 values, 2.28 +/- 0.87 nM and 27 +/- 5 microM respectively. On the other hand, resveratrol inhibited state 4 following a concentration-dependent curve with an EC50 of 37 +/- 11 microM. When complex IV operated alone, resveratrol (100 microM) did not modify oxygen consumption compared with control, indicating that this molecule did not inhibit complex IV. Thus resveratrol inhibits the mitochondrial respiratory chain through complexes I to III. In order to confirm these data, we measured the enzymatic activity of ubiquinol cytochrome c reductase alone and in the presence of resveratrol. In the presence of disrupted mitochondria, after freeze thawing cycles (three times), resveratrol inhibited about 20% of complex III activity. These results suggest that resveratrol and DUQH2 could be competitive on complex III. Resveratrol significantly inhibited ATPase activity (p < 0.001) following a biphasic curve with two EC50 values, 0.39 +/- 0.15 nM and 23.1 +/- 6.4 microM, both representing about 80% of oligomycin-dependent ATPase total activity. Resveratrol was effective as a protecting agent on the three models of oxidation. On lipid peroxidation of brain synaptosomes induced by the Fenton reaction, it was three times more potent than DUQH2. Its effectiveness in reducing 1,1-diphenyl-2-picryl hydrazyl radical (DPPH degrees) showed a stoichiometry of two, indicating that two hydrogen atoms of resveratrol were abstracted by the process. Resveratrol was also able to scavenge the superoxide anion (O2 degrees) generated from rat forebrain mitochondria in a concentration dependent manner. In conclusion, resveratrol can decrease complex III activity by competition with coenzyme Q. This property is especially interesting as this complex is the site where reactive oxygen substances (ROS) are generated. By decreasing the activity of complex III, resveratrol cannot only oppose the production of ROS but can also scavenge them. PMID- 10370870 TI - Resveratrol, map kinases and neuronal cells: might wine be a neuroprotectant? AB - Alcohol is noxious to the brain and peripheral nervous system. However, wine contains substances that may have positive biological and pharmacological effects. Resveratrol is the most studied and probably the most active of these substances. This naturally occurring compound, which is present in wine and grapes, reduces oxidative stress in neuronal-like cell cultures. We have shown that resveratrol induces phosphorylation of the mitogen-activated protein (MAP) kinase family members, extracellular regulated kinase 1 (ERK1) and ERK2, in the human neuroblastoma SH-SY5Y cells in vitro at much lower concentrations than those found in the plasma of rats after oral wine administration. MAP kinases are involved in numerous different aspects of signal transduction in the cells. In particular, phosphorylation of ERK2 has been related to the synaptic changes at the basis of memory and learning processes. These findings, together with our own, on resveratrol-induced activation of MAP kinases in human neuronal-like cells, and previously published epidemiological studies which have demonstrated an inverse relationship between moderate wine intake and dementia, suggest that wine (not alcohol) may have a positive effect on nervous cells. PMID- 10370871 TI - Stimulation of endogenous adenosine release by oral administration of quercetin and resveratrol in man. AB - Epidemiological evidence indicates that moderate alcohol consumption is associated with a significant decrease in the incidence of certain cardiovascular disorders, which can lead to impaired quality of life and to death. However, there are no objective data suggesting a cause-effect relationship and detailed research based on definitive working hypotheses is needed. We tested two flavonoids in man and found that these substances can belong, at least in part, to a wine-dependent mechanism, which leads to increased adenosine plasma levels. If these results could be confirmed by analyzing all the possible influences leading to blood nucleoside increase, a hypothesis of diet-dependent cellular preconditioning could be discussed. PMID- 10370872 TI - Nonalcoholic compounds of wine: the phytoestrogen resveratrol and moderate red wine consumption during menopause. AB - The literature on the natural sources of resveratrol, a phytochemical substance found in grapes and wine, is reviewed herein. Its structure is similar to that of diethylstilbestrol and, like other authors, we consider that resveratrol might be a phytoestrogen. We analyzed the populations who ingest this substance, as well as the known biological effects of phytoestrogens in humans. The literature on the effects of resveratrol on female reproduction, osteoporosis and cancer was assessed using relevant case reports and cohort studies, as well as randomized trials and review articles. We conclude that phytoestrogens exhibit physiological effects in humans and that these estrogenic actions increase the biological reactions produced by moderate red wine consumption with meals. PMID- 10370874 TI - Beneficial effects of white wines. AB - Wine phenolics have been reported to have health benefits, including protection against cardiovascular diseases and anticarcinogenic effects. White wines are usually made with the free-running juices without pomace, which has no contact with the grape skins. This is the main reason that the phenolic content of white wines is lower than that of red wines. However, white wine phenols have a comparable or higher antioxidant capacity than red wine phenols. Therefore, it is important to determine which phenolic compounds are present in white wines and which factors affect phenolic composition. We studied the influence of several factors, including variety and different technological processes, on phenolic composition. Significant differences were observed when any of these variables were considered. Consequently, if wine makers were to take these factors into consideration, the possible beneficial effects of their wines could be increased. PMID- 10370873 TI - Cardioprotection of red wine: role of polyphenolic antioxidants. AB - Epidemiological studies suggest that the consumption of wine, particularly of red wine, reduces the incidence of mortality and morbidity from coronary heart disease. This has given rise to what is now popularly termed the "French paradox". The cardioprotective effect has been attributed to antioxidants present in the polyphenol fraction of red wine. Grapes contain a variety of antioxidants, including resveratrol, catechin, epicatechin and proanthocyanidins. Of these, resveratrol is present mainly in grape skin while proanthocyanidin is present in the seeds. In this report, we provide evidence that red wine extract as well as resveratrol and proanthocyanidins are equally effective in reducing myocardial ischemic reperfusion injury, which suggests that these red wine polyphenolic antioxidants play a crucial role in cardioprotection. PMID- 10370875 TI - Wine and polyphenols related to platelet aggregation and atherothrombosis. AB - Epidemiological studies have demonstrated an inverse correlation between moderate wine and alcohol consumption and morbidity and mortality from coronary heart disease (CHD). This protective effect has been associated with an increase in the plasma level of high density lipoprotein (HDL)-cholesterol, as it is well known that plasma HDL is inversely correlated with CHD. In addition, it has become evident that blood platelets contribute to the rate of development of atherosclerosis and CHD through several mechanisms. Recent studies have shown HDL cholesterol levels can explain only 50% of the protective effect of alcoholic beverages. The other 50% may be partly related to decreased platelet activity. The antiplatelet activity of wine is explained not only by ethanol but also by the polyphenolic components with which red wines are richly endowed. Several studies carried out in humans and animals have shown that wine phenolics could exert their effects by reducing prostanoid synthesis from arachidonate. In addition, it has been suggested that wine phenolics could reduce platelet activity mediated by nitric oxide. Moreover, wine phenolics increase vitamin E levels while decreasing the oxidation of platelets submitted to oxidative stress. However, a rebound phenomenon of hyperaggregability is observed after acute alcohol consumption but not after wine consumption. This protection afforded by wine has been duplicated in animals with grape phenolics added to alcohol. This rebound phenomenon could explain ischemic strokes or sudden deaths known to occur after episodes of drunkenness. It appears that wine and wine phenolics in particular could significantly inhibit platelet aggregation and that this could explain, at least in part, the protective effect of red wine against atherosclerosis and coronary heart disease. PMID- 10370877 TI - Wine: risk factors for liver disease and antifibrotic compounds. AB - Wine can be considered an integral part of the Mediterranean diet and moderate alcohol intake can be beneficial for health. This health-promoting effect is presumably due to the presence of antioxidant substances. It is also known that excessive alcohol intake can lead to liver cirrhosis. A main pathogenetic mechanism in liver cirrhosis is the activation of hepatic stellate cells which acquire a myofibroblast-like phenotype. Excessive production of oxidative stress products may initiate the activation process. Phenolic compounds contained in red wine have been shown to have antifibrotic properties on activated hepatic stellate cells. In vitro and in vivo studies are needed for a better evaluation of the clinical relevance of these findings. PMID- 10370878 TI - Wine and migraine: compatibility or incompatibility? AB - According to popular belief, alcoholic beverages are to be avoided in the case of headache, a term which includes migraine, the most common type of headache. An imbalance between pain transmission and inhibition has been suggested and partly proved to be the mechanism of migraine. This means that peripherally acting substances following wine intake are unlikely to trigger migraine attacks. We hypothesized that factors other than the mere consumption of alcohol can trigger migraine attacks. In an attempt to corroborate this assumption, we carried out a 14-month study in 307 volunteers. All the volunteers had no health problems apart from suffering from migraine without aura. During the entire study period, patients had to complete a diary/questionnaire every time they consumed alcohol. The questionnaires included items regarding the quantity (measured in dl) and the type of alcohol they consumed as well as information about their lifestyle. The volunteers also had to complete a pain diary. It was observed that spirits and sparkling wines were significantly (p > 0.0001) more frequently related to migraine attacks than other alcoholic beverages. Nonetheless, there was no statistical relationship between the consumption of alcohol and migraine attacks. On the other hand, a positive relationship was established between stressful events and the onset of migraine attacks. As an overall result, it was observed that low amounts of alcohol (i.e., 1 dl of 4-14% alcohol/vol. and 0.4 dl of 35 42% alcohol/vol.) did not induce a significant increase in the frequency of migraine attacks. Moreover, it emerged that alcoholic beverage intake during stress periods was related to a significantly higher frequency of migraine attacks (p > 0.0001 for spirits and sparkling wines, p > 0.009 for red wine and p > 0.006 and p > 0.004 for white wine and beer, respectively). Routine blood tests revealed that the subjects who prefer red wine showed a lower level (p > 0.05) of total cholesterol, independently of sex or age. Due to the small sample size (197 vs. 96 tested subjects), this last observation can not be regarded as conclusive. PMID- 10370876 TI - Plasma polyphenols and antioxidants, oxidative DNA damage and endothelial function in a diet and wine intervention study in humans. AB - An intervention study was performed to evaluate the influence of a Mediterranean diet, a high fat diet, and their supplementation with red wine in moderate amounts, on biochemical, physiological, and clinical parameters related to atherosclerosis and other chronic diseases. For 3 months two groups of 21 male volunteers each, received either a Mediterranean diet or a high fat diet; during the second month, red wine was added isocalorically, 240 ml/day. Participants were kept under close medical and nutritional surveillance. At days 0, 30, 60 and 90, clinical, physiological and biochemical evaluations were made. Plasma vitamin C was significantly decreased in the high fat diet group compared to the Mediterranean diet group. After wine supplementation to the Mediterranean diet, a significant 13.5% increase in plasma vitamin C was observed. Furthermore, when wine was added vitamin E decreased significantly in plasma, 15% in the high fat diet and 26% in the Mediterranean diet. Total plasma antioxidant capacity (total antioxidant reactivity) increased 28% above basal levels in the Mediterranean diet group, but not in the high fat diet group. In both groups, wine induced a marked increase in total antioxidant reactivity above basal levels, 56% and 23%, respectively. Oxidative DNA damage, detected as 8-hydroxydeoxyguanosine (8-OHdG) levels in blood leukocyte DNA, was markedly increased by the high fat diet; however, it was strongly reduced, to approximately 50% basal values, after wine supplementation, both in the high fat diet and Mediterranean diet groups. Endothelial function, evaluated noninvasively as flow-mediated vascular reactivity of the brachial artery, was suppressed by the high fat diet, and was normal after wine supplementation. These effects are attributed to oxidative stress associated with a high fat diet, and to the elevated plasma antioxidant capacity associated with wine consumption and the Mediterranean diet. The results presented support the following conclusions: a high fat diet induces oxidative stress; a diet rich in fruits and vegetables enhances antioxidant defenses; wine supplementation to a high fat or a Mediterranean diet increases plasma antioxidant capacity, decreases oxidative DNA damage, and normalizes endothelial function. PMID- 10370879 TI - Scientific basis for the health benefits of the Mediterranean diet. AB - The meaning and influence of the recently introduced concept of the "Mediterranean diet" are explained herein. Although now viewed as a discovery, the Mediterranean people's surprise at this "new" concept is stressed as they are already well acquainted with the qualities of this dietary model. Important changes in lifestyle and nutrition are currently taking place, which may not be the best way to maintain a healthy diet. Possibly, too many ancient nutritional traditions have been abandoned. If this is so, the return to this old dietary model, which has proven to be so beneficial to health, should be welcomed. To understand the significance of the Mediterranean diet the most beneficial compounds and details of the Mediterranean "classical model of lifestyle and nutrition" are explained. Finally, hypotheses about the scientific basis of the benefits of the Mediterranean diet are explained. PMID- 10370880 TI - Fragments of tradition: revisiting the virtues of wine. AB - Wine is a characteristic example of how difficult it is to make a clear-cut distinction between products that are good for man's health and those that damage it. Over the centuries wine has not only been used as a foodstuff, a luxury item and a poison, but also a drug, a carrier for preparing medicines, a cure-all for the ills of the body and soul, and has even assumed an aura of holiness and mysticism. All of these can fall into the categories of treatment and prevention. In taking an historic look at the various opportunities wine has provided over the centuries--especially in the history of pharmacy--the aim is not to reject the product, but rather to reconfirm its qualities in the light of recent discoveries, and above all, highlight how these qualities are inherent to the product as a whole, not just in terms of its individual components. Wine has many characteristics that could classify it as a plant compound. This could serve as a reevaluation of tradition and traditional medicines, not by dissecting them, but by reconsidering them in their historical, medical and even religious entireties. Finally, but no less importantly, some active ingredients of wine with acknowledged therapeutic properties have been used for millennia in traditional Chinese plant therapy. Ingredients contained in other plants have been used for the same purposes and have recently been "scientifically" discovered. PMID- 10370881 TI - [Ethics and genetics in psychiatry]. PMID- 10370882 TI - [Somatic complains, psychiatric disorders and MMPI 2]. AB - Somatic complaints are a frequent motive of medical consulting. They are often linked to an underlying psychiatric disorder and may be its only manifestation, and thus may cause misdiagnosis. Links between somatic complaint and the main psychiatric disorders (depression, personality disorders and psychosis) are studied among a sixty-six psychiatric patients sample who answered the 567 questions of the latest Minnesota Multiphasic Personality Inventory (MMPI 2). Correspondence between clinical diagnosis and MMPI 2 self-evaluation is also studied. PMID- 10370883 TI - [Utilization of a self-contained psychometry-oriented device, for study of affective and cognitive function of a subject]. AB - We appraised the possibility to use the Psycho-Log 24, an ambulatory, adapted to chronopsychometry device, which realizes simplified questionnaires and performance test adaptable to each subject; this device has dimensions like a pocketbook; its storage capacity is compatible with a long time use. 9 adult volunteers (8 healthy subjects and 1 depressive patient) participated to this study. All of them effected reaction times (TRS) during 3 to 31 days at the rate of at least 4 sessions/day; some of them completed mental calculations (CM), letters cancellations (BRL) and auto-estimation scales (from 1 to 9 items, also measured by a classic method). For TRS and all the subjects, the frequencies histograms calculated by hour and on a 24 h scale, showed a bimodal variation, with a major peak suggesting a circadian rhythm. 5 subjects had different time for the daily best performances of each hand (shifted from 5 to 12 h). In the healthy adults, for BRL and CM, the best performances in speed and precision are shifted and could be related to the "fatigue" auto-estimated peaks. There is a significant correlation between auto-estimations by Psycho-Log and the classic method. The coherence between the auto-estimations of the depressive subject is strong during the remission stage, and lower during the relapse stage. The used method seems an effective mean to know the temporal structure of the cognitive and affective functions of a subject, and to estimate the preserving of this structure; the method first could constitute an exploratory mean for an objective evaluation of the drugs effects and patients quality of life, secondly could be implicated in researches with diagnosis target. PMID- 10370884 TI - [Anxiolytics and hypnotics use by psychiatric inpatients' children: preliminary study]. AB - Psychotropic drugs are widely used in adulthood in France, and few studies have yet been made concerning children. Literature studies have shown an important consumption on their part, increasing with age and especially for girls. Certain family factors which may prove to determine such use have been pointed out like family habits of toxique use, maternal depression, mother's occupation and family troubles. This consumption also seems related to psychiatric symptomatology in children together with previous use in childhood. This feasibility study will attempt to research the forms of anxiolytics and hypnotics use by children aged 6 to 16, one parent being a psychiatric in-patient. 51 in-patients of the university section have been included, corresponding to 74 children. The investigator look down socio-demographic date as well as anxiolytics and hypnotics use in a lifetime and that of antalgics for the week prior to the interview. The parents' depressive and anxious symptoms were measured by the HAD and the children's psychopathological symptoms by the CBCL. A phone contact was proposed to teenagers 13 to 16 years old. One patient only refused the interview, 7 patients left without filling in the questionnaire. The parents more often than not refused to establish contact with the teenager. Only 5 teenagers out of 22 could actually be interviewed. Whereas 94.1% of parents use psychotropic drugs, only 16.2% of children have used them throughout their lifetime. The consumption of these drugs is not related to age or sex but rather seems to be linked with the children's symptomatology. The development of this study should confirm these results on a wider scale through a different approach of teenagers. PMID- 10370885 TI - [Study of the relationships between self-injurious behavior and pain reactivity in infantile autism]. AB - Autism can be considered as an early general developmental disorder, characterized by problems of social interaction, problems of verbal and non verbal communication, and behavioral or ideational stereotypes. However, within autism we observe a clinical heterogeneity of autistic disorders which suggests the possibility of autistic subtypes. Several authors hypothesize an analgesia among autistic children; this analgesia may be related to self-mutilation found among autistics. The current research had two objectives: 1) to develop and validate evaluation tools for measuring aggression directed towards the self (Yale-Paris Self-Injurious Behavior Scale: YAPA SIB) and pain reactivity (Pre Linguistic Behavioral Pain Reactivity Scale: PLBPRS); instruments appropriate for autistics and capable of showing different behavioral sub-types; 2) to study in 80 autistic children pain reactivity, self-injurious behavior, and their relation in different observational situations. The results show that the scales of self injurious behavior and pain reactivity have good discriminative capacity, good test-retest reliability, and good validity. The results suggest additionally that the apparent decreased pain reactivity observed in autistics does not derive from a real analgesia but from a different mode of pain expression related to difficulties with verbal communication, body representation and certain cognitive disorders (learning disorders, problems representing sensations and emotions, problems establishing cause-effect relationships). Additionally, there is a significant relationship between certain self-injurious behaviors and the apparent reduced pain reactivity. Interpretations of this result are presented and the possible role of stress in autism is discussed. PMID- 10370886 TI - [Mechanisms of nutrient and drug transfer through the blood-brain barrier and their pharmacological changes]. AB - The blood-brain barrier is formed by the endothelial cells of the brain capillaries. Its primary characteristic is the impermeability of the capillary wall due to the presence of complex tight junctions and a low endocytic activity. Essential nutrients are delivered to the brain by selective transport mechanisms, such as glucose transporter and a variety of amino acid transporters. Although most drugs enter the brain by passive diffusion through the endothelial cells depending of their lipophilicity, degree of ionization, molecular weight, relative brain tissue and plasma bindings--some of them can use specific endogenous transporters. In these cases, binding competition on the transporter with endogenous products or nutrients can occur and limit the drug transfer. The blood-brain barrier can be a major impediment for the treatment of diseases of the central nervous system, since many drugs are unable to reach this organ at therapeutic concentrations. Various attempts have been made to overcome the limiting access of drugs to the brain: chemical modification of drugs, development of more hydrophobic analogs or linking an active compound to a specific carrier. Transient opening of the blood-brain barrier has been achieved by intracarotid infusion of hypertonic mannitol solutions or of bradykinin analogs in humans. Another way to increase or decrease brain delivery of drugs is to modulate the P-glycoprotein (P-gp) whose substrates are actively pumped out the cell into the capillary lumen. We actually dispose of many P-gp inhibitors or inducers in order to enhance the therapeutic effects of centrally acting drugs or to decrease central adverse effects of peripheric drugs. PMID- 10370887 TI - [Risperidone and concept of bipolar neuroleptic]. AB - We present a review of the stimulant properties of risperidone, analyzing them according to the model of bipolar neuroleptic proposed in 1979 by Colonna and Petit. The bipolar neuroleptics (pimozide, sulpiride) present stimulant properties at low dose and sedative effects at high dose whereas monopolar neuroleptics (levomepromazine, chlorpromazine) present from the low dose sedative effects. The stimulant properties might arise from a preferential blockade of the presynaptic dopaminergic receptors, giving and enhancement of the dopaminergic transmission. Risperidone might follow the bipolar model; we review the stimulant effects in laboratory animals and in clinical settings, showing exacerbation of manic symptoms and a possible antidepressive effect. These properties could be due to the capacity to increase the cerebral monoamines by the 5-HT2A and alpha 2 antagonism. Risperidone could be used in the treatment of clinical syndromes combining depressive and psychotic symptoms. PMID- 10370888 TI - [Compliance with and tolerance of sustained-release lithium carbonate]. AB - More than 40 years after its efficacy in the management of acute mania was first described, lithium remains first-line drug treatment for bipolar mood disorders. Studies conducted under natural conditions have shown that one of the main causes of failure is associated with poor treatment compliance. The poor compliance rate observed with treatment of this nature currently stands between 18 and 47%. The differences highlighted in the various studies can be explained, at least in part, by the definitions and assessment methods employed in each study. It is generally acknowledged that the longer the evaluation period, the worse the compliance. Certain factors accounting for poor compliance point to the patients themselves whilst others are directly treatment-related, i.e. therapeutic efficacy and tolerance. Compliance with two pharmaceutical dosage forms of lithium carbonate, namely the standard (250 mg) and sustained-release (400 mg) formulations, was assessed under natural conditions in patients satisfying DSM IV diagnostic criteria for bipolar disorder type I, II or non-specified or for major recurrent depression. Evaluation via a "heteroquestionnaire" was conducted retrospectively over 3 months and prospectively over 6 months. Assessments were carried out at baseline and after 3 and 6 months. Patients who responded at a minimum of two evaluations were included in the statistical analysis. Forty-four (44) of the 50 patients selected were included. A global compliance rate of 74% was recorded at 6 months and this was deemed to be excellent. These results corroborate published data relating to the overall good compliance observed with lithium patients. Tolerance and compliance were better in the group receiving the sustained-release formulation (400 mg per tablet). A certain number of factors likely to influence the quality of compliance were measured. No correlation was established between the quality of compliance assessed by the doctor and data relating to the disorder or to the patients'understanding of and attitude towards treatment. Two variables were significantly correlated with the extent of patient assessed compliance, namely those patients displaying better compliance generally started treatment later in life (35.7 years +/- 12.0 compared with 25 years + 15.6; p = 0.03) and took fewer tablets (1.7 tablets/day +/- 1.1 versus 2.7 tablets/day +/- 1.6; p = 0.03). These factors are discussed. PMID- 10370889 TI - [The treatment of social phobias: efficacy of cognitive and and behavioral group therapy]. AB - 55 patients with social phobias were given group cognitive-behavioural therapy. The study protocol comprised three phases: (1) the pre-inclusion phase with 5 individual consultations before group treatment; (2) the group phase itself, comprising 20 weekly sessions of 2 hours each and (3) the post-therapy phase, comprising a 6 month follow-up group session, and 2 evaluation sessions 6 and 12 months after the end of group therapy. The techniques used were assertiveness training and cognitive therapy. Assessments were carried out using the Rathus Assertiveness Schedule (23), Brief standard self rating for phobic patients (Marks) (19) and Beck Depression Inventory Scores (1), at specified times during the study: at the start of the individual phase; at the start, in the middle and at the end of the group phase, as well as at 6 month and 1 year follow-up. The results are interesting. Follow-up was very good, with 54 out of 55 patients finishing the group treatment, and with 50 patients being evaluated at 6 and 12 months. Global analysis of the results showed that at the end of group therapy, there was a statistically significant improvement in the score on the Rathus Assertiveness Schedule, on the social sub-score, on the brief standard self rating for phobic patients (Marks). The improvement in these variables was maintained at 6 and 12 month follow-up. PMID- 10370890 TI - [Anticonvulsants in agitation and behavior disorders in demented subjects. Report of 8 cases]. AB - Aggressive agitation, agitation and insomnia with generalized anxiety are commonly observed in Alzheimer's disease. These symptoms remain a principal problem in the clinical management of elderly patients. Neuroleptics are commonly the selected medication for controlling severe aggression, especially the violent out bursts often seen in demented patients. Their use is frequently complicated by side effects, particularly somnolence and confusion. Valpromide and Carbamazepine have been efficacy alternatives and very well tolerated. We report eight cases of demented patients who presented an agitation and aggressive behaviors and had been treated with Valpromide or Carbamazepine. The patients agitation was well controlled at that point and had no apparent side effects. A combination Valpromide or Carbamazepine with neuroleptics permitted a reduction doses of neuroleptics and their side effects. We think that these behaviors disorders belong to the mood disorders. The symptomatology is modified because an alteration of cognitive faculty. PMID- 10370891 TI - [Preventive role of lithium in self-injurious behavior: a case report]. AB - The authors report a case of 48 years old woman with mental retardation, followed for 30 years. She has conduct disorder and during this period we note a large number of self-injuries. She swallowed broken glasses, safety pins and once jumped from a bridge. The impulsiveness of these self-injuries is the main factor; they are not planned self-attempts. Lots of different treatments have been prescribed (neuroleptics, antidepressant serotonin uptake blockers and others) and it is only with the lithium that we could observe a period of 2 years without self-injuries. This case questions about the way of action of the lithium and about a possible specificity in the preventive action on conduct disorder with self-injurious behavior, on this type of patient. PMID- 10370892 TI - International Incidence of Childhood Cancer, Vol. II. PMID- 10370893 TI - Epidemiology of childhood cancer. PMID- 10370894 TI - Gaucher's disease: a model for modern management of a genetic disease. PMID- 10370895 TI - Haemochromatosis and the liver. PMID- 10370896 TI - The porphyrias. PMID- 10370897 TI - Amyloidosis and the liver. PMID- 10370898 TI - Alpha 1-antitrypsin deficiency. PMID- 10370899 TI - Hepatic granulomas. PMID- 10370900 TI - Members of the European Association for the Study of the Liver, 1999. PMID- 10370902 TI - Pathogenesis and preclinical course of Parkinson's disease. AB - Idiopathic parkinsonism (IP) is defined by its classic symptomology, its responsiveness to therapies which elevate dopamine levels, and by the failure to identify a specific etiological factor. The progressive and irreversible degeneration of dopaminergic neurons projecting from the substantia nigra pars compacta (SNc) to the striatum and the presence of SNc Lewy bodies are regarded as the essential pathological bases of IP, but neither the initiator(s) nor the nature of the degeneration have been determined, nor its relationship with degenerative changes in other parts of the IP brain. This paper discusses the various hypotheses that have been proposed to explain these phenomena, arguing that IP be regarded as a multisystem disorder, both at the level of individual neurons and at the whole brain level. It is probable that IP is the result of a multifactorial process, and that a cascade of interacting and overlapping biochemical mechanisms determine the course of the disease. PMID- 10370901 TI - Post mortem studies in Parkinson's disease--is it possible to detect brain areas for specific symptoms? AB - Parkinson's disease (PD) is characterized by progressive neuronal loss associated with Lewy bodies in many subcortical nuclei leading to multiple biochemical and pathophysiological changes of clinical relevance. Loss of nigral neurons causing striatal dopamine deficiency is related to both the duration and clinical stages (severity) of the disease. The clinical subtypes of PD have different morphological lesion patterns: a) The akinetic-rigid type shows more severe cell loss in the ventrolateral part of substantia nigra zona compacta (SNZC) that projects to the dorsal putamen than the medial part projecting to caudate nucleus and anterior putamen, with negative correlation between SNZC cell counts, severity of akinesia-rigidity, and dopamine loss in the posterior putamen. Reduced dopaminergic input causes overactivity of the GABA ergic inhibitory striatal neurons projecting via the "indirect loop" to SN zona reticulata (SNZR) and medial pallidum (GPI) leading to inhibition of the glutamatergic thalamo cortical motor loop and reduced cortical activation. b) The tremor-dominant type shows more severe neuron loss in medial than in lateral SNZC and damage to the retrorubral field A8 containing only few tyrosine hydroxylase and dopamine transporter immunoreactive (IR) neurons but mainly calretinin-IR cells. A8 that is rather preserved in rigid-akinetic PD (protective role of calcium-binding protein?) projects to the matrix of dorsolateral striatum and ventromedial thalamus. Together with area A10 it influences the strial efflux via SNZR to thalamus and from there to prefrontal cortex. Rest tremor in PD is associated with increased metabolism in the thalamus, subthalamus, pons, and premotor cortical network suggesting an increased functional activity of thalamo-motor projections. In essential tremor, no significant pathomorphological changes but overactivity of cerebello-thalamic loop have been observed. c) In the akinetic rigid forms of multisystem atrophy, degeneration is more severe in the lateral SNZC with severe loss of calbindin-IR cells reflecting initial degeneration of the striatal matrix in the caudal putamen with transsynaptic degeneration of striatonigral efferences that remain intact in PD. This fact and loss of striatal D2 receptors--as in advanced stages of PD--are reasons for negative response to L dopa substitution. These data suggest different pathophysiological mechanisms of the clinical subtypes of PD that have important therapeutic implications. d) Involvement of extranigral structures in PD includes the mesocortical dopaminergic system, the noradrenergic locus coeruleus, dorsal vagal nucleus and medullary nuclei, serotonergic dorsal raphe, nucleus basalis of Meynert and other cholinergic brainstem nuclei, e.g. Westphal-Edinger nucleus (controlling pupillomotor function), posterolateral hypothalamus and the limbic system, e.g. amygdaloid nucleus, part of hippocampal formation, limbic thalamic nuclei with prefrontal projections, etc. Damage to multiple neuronal systems by the progressing degenerative process causing complex biochemical changes may explain the variable clinical picture of PD including vegetative, behavioural and cognitive dysfunctions, depression, pharmacotoxic psychoses, etc. Future comparative clinico-morphological and pathobiochemical studies will further elucidate the pathophysiological basis of specific clinical symptoms of PD and related disorders providing a broader basis for effective treatment strategies. Parkinson's disease (PD) is characterized by progressive degeneration of the nigrostriatal dopaminergic system and other subcortical neuronal systems leading to striatal dopamine deficiency and other biochemical deficits related to the variable clinical signs and symptoms of the disorder. (ABSTRACT TRUNCATED) PMID- 10370903 TI - Clinical efficacy of budipine in Parkinson's disease. AB - The lipophilic t-butyl analog of 1-alkyl-4,4-diphenyl piperidine, budipine, possesses a polyvalent spectrum of mechanisms of action. Budipine experimentally increased the brain content of norepinephrine, serotonine, dopamine and histamine in reserpine treated rats. Budipine did not alter the receptor affinity of these neurotransmitters but antagonizes the effect of NMDA at its receptor binding site in vitro. Budipine reduced MPP+ toxicity in the nigrostriatal system of mice. This complex pharmacologic profile is not comparable to the one of convenient antiparkinsonian drugs. In clinical trials budipine reduced tremor, akinesia and rigidity. Budipine induced a relevant additional positive effect in patients with an optimal dopaminergic therapy based on levodopa and dopamine agonists, such as bromocriptine. Current available data suggest that the need for levodopa application in early stages of the disease may be postponed by budipine and that the long-term application of budipine may induce a levodopa-sparing effect. PMID- 10370904 TI - Multiple mechanisms of action: the pharmacological profile of budipine. AB - Four major components of the mechanism of action have been identified for the antiparkinsonian drug budipine up to now. 1) The primary action of budipine is an indirect dopaminergic effect as shown by facilitation of dopamine (DA) release, inhibition of monoamine oxidase type B (MAO-B) and of DA (re) up-take and stimulation of aromatic L-amino acid decarboxylase (AADC), which in sum might be responsible for enhancing the endogenous dopaminergic activity. 2) Radioligand and functional studies at the N-methyl-D-aspartate (NMDA) type glutamate receptor characterize budipine as a low-affinity, uncompetitive antagonist with fast kinetics and moderate voltage-dependency at the phencyclidine (PCP) binding site, comparable to that observed with amantadine, thereby counteracting an increased excitatory glutamatergic activity. 3) The antimuscarinic action of budipine, verified by functional and binding studies at native muscarinic M1-M3 and human recombinant m1-m5 receptor subtypes in vitro, is up to 125-fold weaker than that of biperiden and corresponds to its approximately 100-fold lower potency to cause experimentally-induced peripheral antimuscarinic effects and explains only part of its high potency, which equals biperiden, to suppress cholinergically evoked tremor. 4) An additional inhibition of striatal gamma-aminobutyric acid (GABA) release by budipine may be beneficial to suppress an increased striatal GABAergic output activity. The contribution of other observed effects to the therapeutic action of budipine, i.e. weak stimulation of noradrenaline and serotonin release, binding to brain sigma1 receptors and blockade of histamine H1 receptors, is not yet clear. By means of these multiple mechanisms, budipine might correct the imbalance of striatal output pathways by restoring DA levels in the striatum, and positively influence the secondary changes in other transmitter systems (glutamate, acetylcholine, GABA) observed in Parkinson's disease. PMID- 10370905 TI - Parkinson's disease: one disease entity or many? AB - Although most neurologists accept that Parkinson's disease (PD) is a unique disorder, we still lack a valid diagnostic biological tool and therefore depend primarily on clinical examination in diagnosis. The distinction between PD and other parkinsonian syndromes is discussed. PMID- 10370906 TI - Parkinson's disease--a multifactorial neurodegenerative disorder. AB - The pathogenesis of idiopathic Parkinson's disease (PD) is not known, but is thought to be multifactorial, deriving from environmental factors acting on genetically predisposed individuals with aging. Association studies of DNA polymorphisms are able to detect a genetic background predisposing to PD. Mechanisms as oxidative stress, xenobiotica toxicity and altered dopamine metabolism might lead to a selective cell death of most vulnerable nerve cells and represent the primary subject to be studied by DNA analysis. Furthermore, protein aggregation is likely to be a major cause for the disease. Recently it has been shown that alpha-synuclein is accumulated in Lewy bodies of sporadic PD and mutated in some rare families with an autosomal dominant trait of the disease (ADPD). The identification of further genes responsible for PD will subsequently lead to first insights into the pathogenesis of one of the most common neurodegenerative disorders in humans. PMID- 10370907 TI - Mechanism and consequences of nerve cell death in Parkinson's disease. AB - The etiology of Parkinson's disease remains unknown, making it difficult to develop therapeutical approaches to stop the progression of the disease. The best known treatment to date is based on the use of L-DOPA or dopaminergic agonists. These are merely substitutive therapies and have limitations because of their side effects. Thus, the development of new therapeutical strategies will require a far better knowledge of the mechanism and the consequences of nerve cell death in Parkinson's disease. Parkinson's disease is characterized by a selective vulnerability of sub-populations of dopaminergic neurons in the mesencephalon. The fact that the neurons which degenerate in Parkinson's disease are already sensitive to oxidative stress in control subjects and the reported increased production of oxygen free radicals in Parkinson's disease suggest that oxidative stress may be involved in the mechanism of nerve cell death. Furthermore, oxygen free radicals are also involved in an oxygen-dependent pro-apoptotic pathway stimulated by the inflammatory reaction observed in Parkinson's disease. These data suggest that anti-oxidant or anti-inflammatory treatments may slow down the progression of the disease. On the other hand, new substitutive therapies may be developed by trying to restore the activity of the neurons located downstream from the nigrostriatal pathway. Indeed, the nigrostriatal denervation induces a hyper-activity of the output structures of the basal ganglia (internal segment of the globus pallidus and substantia nigra pars reticulata), as demonstrated in various animal models of the disease. These changes in the activity of the output structures of the basal ganglia seem to be directly induced by the hyperactivity of the glutamatergic afferent fibers from the subthalamic nucleus. The fact that L-DOPA treatment or a reduction in the activity of the subthalamic nucleus alleviate the symptoms of the disease and restore the activity of the output structures of the basal ganglia in parkinsonism suggests that these structures play a key role in the pathophysiology of the disease and could represent a potential therapeutic target. PMID- 10370908 TI - Functional imaging of Parkinson's disease: is it possible to detect brain areas for specific symptoms? AB - H2(15)O PET activation studies enable the brain systems involved in controlling different aspects of motor function to be defined. In Parkinson's disease (PD) freely chosen limb movements are performed slowly. This bradykinesia is associated with selective underactivity of the supplementary motor area and dorsal prefrontal cortex, frontal association areas that receive subcortical input principally from the basal ganglia. At the same time there is compensatory overactivity of the lateral premotor and parietal cortex, areas that have a primary role in facilitating motor responses to visual and auditory cues. This finding explains why PD patients find it easier to perform cued as opposed to freely chosen actions. Levels of activation of the supplementary motor area and dorsal prefrontal cortex in PD can be restored with dopaminergic medication, implants of fetal mesencephalic tissue, internal pallidotomy or high frequency electrical subthalamic stimulation. Activation studies suggest that Parkinsonian rest tremor arises from a combination of inappropriate overactivity of cerebellar connections and loss of dopaminergic function. When tremor is relieved by ventral thalamotomy or thalamic stimulation this cerebellar overactivity is corrected but at the expense of reducing levels of primary motor cortex activation. It has been hypothesised that dyskinesias in PD arise due to altered dopamine receptor binding following chronic exposure to levodopa stimulation. Functional imaging findings, however, are against this hypothesis and rather suggest that downstream increases in basal ganglia opioid neurotransmission are more likely to be relevant. PMID- 10370909 TI - Multiple system atrophy. AB - Multiple system atrophy (MSA) is a degenerative central nervous system disease of unclear origin. Patients affected typically show symptoms attributable to the combined involvement of the extrapyramidal, pyramidal, cerebellar and autonomic nervous systems. At onset patients mostly see a doctor because of extrapyramidal or--more rarely--cerebellar symptoms. Evidence of autonomic nervous system involvement is often not apparent, at least to the neurologist, before the history is taken. In later stages, by contrast, involvement of all of the above systems is clinically detectable. PMID- 10370910 TI - Tremorlytic activity of budipine in Parkinson's disease. AB - In order to objectively quantify the tremorlytic activity of budipine in Parkinson's disease (PD) we performed longterm tremor recordings in a subset of patients enrolled in two clinical trials. Eleven PD patients with marked tremor participating in an open-label study underwent longterm recording before and during medication. Nine patients completed the study. Tremor occurrence was reduced from 52 +/- 18.6% to 34.7 +/- 19.3% (p < 0.05); tremor intensity decreased from 15.3 +/- 4.8 (SNR) to 11.3 +/- 4.8 (p < 0.01). UPDRS tremor subscores were also significantly improved. Fourteen patients who enrolled in a multicenter, double-blind, placebo-controlled study underwent longterm tremor analysis in addition to the Columbia University Rating Scale (CURS). Tremor occurrence was improved in the budipine group (n = 7) from 24.7 +/- 15.5% to 14.8 +/- 14.5% (p < 0.05). Tremor intensity decreased from 9.1 +/- 2.5 (SNR) to 7.2 +/ 1.6. However, the latter result was statistically not significant, probably due to the small patient number. In the placebo-group (n = 7) there was no reduction of tremor occurrence or of tremor intensity. The CURS sum score was improved from 5.7 to 3.0 in the budipine group, whereas there was only a smaller improvement in the placebo group (from 7.1 to 5.5). These data suggest that budipine is an effective tremorlytic agent in PD, which may be used as an alternative to anticholinergics. PMID- 10370911 TI - Current management of motor fluctuations in patients with advanced Parkinson's disease treated chronically with levodopa. AB - Motor fluctuations after long-term administration of levodopa may be due to central pharmacodynamic mechanisms such as reduced striatal synthesis and storage of dopamine from exogenous levodopa and subsensitization of postsynaptic dopaminergic receptors. Peripheral pharmacokinetic mechanisms may be equally important, particularly in motor fluctuations of the "delayed on" (increased time latencies from dose intake to start-up of clinical benefit) and "no-on" (complete failure of a levodopa dose to exert an "on" response) types. Levodopa itself has a very poor solubility. In addition, there is delayed gastric emptying in many advanced patients. Therefore, an oral dose of levodopa may remain in the stomach for long periods of time before it passes into the duodenum where there is immediate absorption. Consequently, in order to overcome response fluctuations caused by impaired pharmacokinetic mechanisms and to improve its absorption, we recommend that levodopa be taken in multiple small doses, on an empty stomach, preferably crushed and mixed with a lot of liquid. Protein intake should be minimized. Prokinetic drugs such as prepulsid (Cisaprid) could be used to facilitate gastric motility and levodopa transit time. Administration of crushed levodopa through nasoduodenal or gastrojejunostomy tubes may be helpful in certain circumstances. Bypassing the stomach with subcutaneous injections of apomorphine may provide dramatic rescue from difficult "off" situations. Oral and s.c. administration of novel, extremely soluble prodrugs of levodopa, e.g., levodopa ethylester, may offer a new approach to overcome difficulties in levodopa absorption. Addition of dopamine agonists, MAO-B inhibitors, COMT inhibitors and controlled release levodopa preparations may be helpful in prolonging the duration of efficacy of each single levodopa dose. Levodopa, administered orally, usually combined with peripheral dopa decarboxylase inhibitors, continues to be the most widely-used and most effective pharmacological treatment for Parkinson's disease (Melamed, 1987). Undoubtedly, the outstanding therapeutic success of levodopa represents a dramatic and revolutionary breakthrough in medicine, in general, and in neurology, in particular. Although, since the introduction of levodopa, there have been many additional pharmacological and even surgical anti-parkinsonian strategies, it still stands out as a mandatory axis of treatment in the majority of patients (Steigler and Quinn, 1992). Indeed, levodopa therapy improves, sometimes markedly, the motor signs and symptoms of the illness, the functional capacity and quality of life and perhaps also life expectancy of the afflicted patients. It is therefore unfortunate that after an initial problem-free period of successful, smooth and stable clinical benefit from levodopa that lasts about two to five years, the responsiveness of many patients worsens with the emergence of a variety of complications (Marsden et al., 1982; Hardie et al., 1984). These adverse reactions include dyskinesias and dystonias, psychotic problems and, particularly, the troublesome motor fluctuations (Marsden and Parkes, 1977; Marsden, 1994). The latter phenomenon may be particularly complex, limiting and disabling. It is believed that most patients on long-term levodopa therapy will, sooner or later, develop response fluctuations of varying types and severity (Riley and Lang, 1993). Because of the serious impact of these phenomena on the quality of life and function of the patients, many efforts are now being undertaken to identify the responsible mechanisms and to devise preventive and therapeutic measures. PMID- 10370912 TI - Differentiation of dopamine agonists and their role in the treatment of Parkinson's disease. AB - Since the pioneering work of Hornykiewicz and his colleagues, it has been recognized that dopaminergic cells die selectively in Parkinson's disease, and considerable improvement in symptoms can be achieved by administering levodopa, so that it may be converted to dopamine. However, levodopa has side-effects, and its duration of action is relatively brief. For these reasons, alternative approaches have been undertaken to stimulate the dopamine receptors. In particular, artificial agonists for dopamine receptors have been developed. The pioneer compound was bromocriptine, which stimulates the D2 family of receptors. Bromocriptine is an ergot derivative, and other compounds that are structurally related to ergot have been developed. In particular, lisuride and pergolide have been used for several years. Recently, an ergot derivative with an exceptionally long plasma half-life has been studied, cabergoline. Now there are also non-ergot derivatives that are D2 agonists, and are likely to have a role in the treatment of Parkinson's disease. Both ropinirole and pramipexole fall into this category, and each has been released in various countries for the treatment of Parkinson's disease. All of these compounds stimulate the D2 family of receptors, but they have varying actions on the D1 family of receptors. At present, there is no definite information on the role of the D1 family of receptors in either the therapeutic response to levodopa, or the development of adverse reactions. However, preliminary studies with a D1 agonist, ABT-431, are now in progress. PMID- 10370913 TI - Free radical scavengers: chemical concepts and clinical relevance. AB - Free radicals are involved in the pathology of many CNS disorders, like Parkinson's disease, Alzheimer's disease, or stroke. This discovery lead to the development of many radical scavengers for the clinical treatment of neurodegenerative diseases. In this review, the different chemical concepts for free radical scavenging will be discussed: nitrons, thiols, iron chelators, phenols, and catechols. Especially catechols, like the naturally occurring flavonols, the synthetic drug nitecapone, or the endogenous catacholamines and their metabolites, are of great interest, as they combine iron chelating with radical scavenging activity. We present data on the radical scvenging activity of dopamine and apomorphine, which prevent lipid peroxidation in rat brain mitochondria and protect PC12 cells against H2O2-toxicity. PMID- 10370914 TI - Differential diagnosis of tremor. AB - The differential diagnosis of tremor is based on the clinical distinction of rest, postural and intention tremor and the presence of additional clinical signs and data from the medical history. The most common pathological tremors are essential tremor and the tremors of Parkinson's disease. Among the patients with essential tremor those with intention tremor are often misdiagnosed as cerebellar tremors. Patients with monosymptomatic resting tremors represent a special subgroup of Parkinson's disease. Primary orthostatic tremor and dystonic tremor are rare clinical syndromes which have recently been well defined. Holmes' tremors are defined by their low frequency and the occurrence of resting and intention tremor. Palatal tremor can be separated into two subgroups. Psychogenic tremor can be diagnosed on the basis of clinical criteria. The gold standard of tremor differential diagnosis is still based on clinical criteria. PMID- 10370916 TI - Effects of a thermoplastic foot orthosis on patellofemoral pain in a collegiate athlete: a single-subject design. AB - STUDY DESIGN: Single-subject, A-B-A-B format. OBJECTIVE: To evaluate the effects of a foot orthosis on pain and function by controlling forefoot pronation. BACKGROUND: Foot orthoses have been recommended to treat patellofemoral pain by controlling excessive pronation, but few studies have used an experimental design to validate this approach. METHODS AND MEASURES: A 19-year-old collegiate softball player had an acute onset of patellofemoral pain in her left knee. A visual analog scale was used to assess pain intensity and a functional index questionnaire (FIQ) was used to measure the subject's perceived functional level. RESULTS: Statistical analysis using the celeration line technique showed significant improvements between all the consecutive baseline and intervention phases of the VAS data and in 7 of the 9 paired phases of the FIQ data. CONCLUSIONS: Limiting pronation with a thermoplastic orthosis was an effective treatment for this subject. PMID- 10370915 TI - Relationship between static and dynamic foot postures in professional baseball players. AB - STUDY DESIGN: Observational study of static and dynamic foot postures in professional baseball players. BACKGROUND: Throughout the course of a professional baseball season, running, cutting, and sprinting activities can produce a breakdown in players' foot function, causing overuse injuries. OBJECTIVES: To investigate the relationship between static and dynamic foot postures; to determine the occurrence of abnormal foot postures in professional baseball players and the incidence of overuse injuries in the lower extremity; and to compare the foot postures of pitchers to those of positional players. METHODS AND MEASURES: The foot postures of 74 professional baseball players were evaluated at rest and during gait. Measures of static foot posture were obtained with a goniometer and included the subtalar neutral position, forefoot/rearfoot position, ankle joint dorsiflexion, tibial angle in standing, and calcaneal angle in standing. The FootTrak motion analysis system provided measures of dynamic foot posture (rearfoot supination and pronation) during the stance phase of gait. A questionnaire was completed by players who reported previous lower extremity injuries. The chi-square statistic was used to determine the associations between forefoot position (varus or valgus) and the amount of foot pronation during gait. RESULTS: The forefoot varus and calcaneal valgus in standing was significantly associated with the maximum pronation during the stance phase of gait. Of the 65 players who demonstrated excessive pronation (> 8 degrees), 28 (43%) also reported a previous lower extremity injury. No statistically significant difference occurred, however, between injured and uninjured players with respect to the mean values of static or dynamic foot posture. In addition, foot postures were not associated with a player's position. CONCLUSIONS: Selected measures of static rearfoot and forefoot postures may have value in predicting dynamic rearfoot movement during the stance phase of gait. Excessive pronation in the baseball players we studied was not found to be a significant contributing factor in the development of overuse injuries. PMID- 10370917 TI - Relationship between ankle invertor H-reflexes and acute swelling induced by inversion ankle sprain. AB - STUDY DESIGN: Single group, post-test design using the uninvolved lower extremity as the experimental control. OBJECTIVES: To determine relationships between ankle swelling and flexor digitorum longus and peroneus longus H-reflex amplitude and latency. BACKGROUND: Primary capsuloligamentous injury, neural injury, and joint effusion and swelling may contribute to H-reflex changes following inversion ankle sprain. The relationship between ankle swelling and invertor or evertor H reflexes has not been reported. METHODS AND MEASURES: Fifteen subjects with acute grade I or II inversion ankle sprains (mean +/- SD) 6.5 +/- 3 days after onset participated in this study. Swelling was estimated using a tape measure and the figure-of-eight girth assessment method. H-reflexes were determined using standard techniques. Paired t-tests were used to compare mean differences in ankle girth (swelling) and ankle invertor or evertor H-reflex amplitude and latency between the involved and uninvolved limbs. Pearson product moment correlations were used to assess relationships between swelling and H-reflex variables. RESULTS: Involved limb ankle girth was increased with respect to the uninvolved limb (1.5 +/- 0.9 cm) and the involved ankle flexor digitorum longus latency was delayed (0.72 +/- 0.7 ms). There was a moderate positive association (r = 0.73) between the latency delay in the involved ankle flexor digitorum longus and swelling. There were no significant differences in H-reflex amplitude and peroneus longus latency between ankles. CONCLUSIONS: Grade I or II inversion sprains and the related swelling appear to delay involved ankle flexor digitorum longus latency to a greater extent than peroneus longus latency. Clinicians need to direct greater attention to the ankle invertors when designing and implementing ankle rehabilitation programs, particularly during the swelling management phase of treatment. PMID- 10370918 TI - Lumbosacral position sense during pelvic tilting in men and women without low back pain: test development and reliability assessment. AB - STUDY DESIGN: A single group test-retest design to evaluate the reproducibility of lumbosacral position sense measurements. OBJECTIVES: To develop a measure of position sense in the lumbosacral area and to determine test-retest reliability. BACKGROUND: Proprioception, muscle control, and coordination training could be the key issues in resolving neuromuscular dysfunction in patients with low back pain, but there are no standard ways to assess these parameters. METHODS AND MEASURES: A piezoresistive accelerometer attached to the skin over the sacrum was used to research the repositioning accuracy of active pelvic tilting, between days, of 14 young nonimpaired subjects (20 to 26 years of age) in standing. RESULTS: The mean absolute error for repositioning accuracy (the difference between criterion and matching positions) was 1.81 degrees (+/- 0.85). The intraclass correlation coefficient between measurements obtained on days 1 and 2 was moderate (R = 0.51). The average standard error of measurement associated with the intraclass correlation coefficient was 0.5 degree (95% confidence interval = +/- 0.99 degree). CONCLUSIONS: These findings suggest that the proposed test is sensitive with moderate test-retest reliability to examine lumbosacral position sense in healthy subjects. Further adjustments in the testing protocol are needed to improve the test-retest reliability. PMID- 10370919 TI - Effects of instruction in jumping technique and experience jumping on ground reaction forces. AB - STUDY DESIGN: Randomized, controlled trial. OBJECTIVE: To determine the roles that augmented feedback from instruction in jumping technique and sensory feedback from experience jumping play in assisting individuals to land softly from a jump. BACKGROUND: Jumping and landing activities play a major role in many sports and daily activities. Feedback may assist individuals in decreasing landing forces and thus reduce the chances of sustaining an injury. METHODS AND MEASURES: Nonimpaired subjects (n = 91) were randomly assigned to either an augmented or sensory feedback condition. All subjects were asked to jump from a box 300 mm in height and land as softly as possible on a force plate. Pre intervention ground reaction forces (GRF) were recorded. Subjects in the augmented feedback condition were then given instructions to focus on hip and knee joint motion as well as a forefoot landing technique for their next jump. Subjects in the sensory feedback condition were asked to use the experience of their first jump to land softly for their next jump. Post-intervention GRFs were recorded and all GRFs were expressed as a multiple of body weight. RESULTS: Those in the augmented condition significantly reduced their GRF scores from pre-(mean = 4.53 +/- 1.51) to post-(mean = 3.57 +/- 1.10) jump, whereas those in the sensory condition did not (mean pre GRF = 4.51 +/- 1.77; mean post GRF = 4.33 +/- 1.54). CONCLUSIONS: High ground reaction forces may be a precipitating factor associated with an injury, where the site of tissue damage would benefit from decreased forces. These findings support the use of instructions related to joint motion to reduce landing forces. PMID- 10370921 TI - Update: NCCN practice guidelines for the treatment of breast cancer. National Comprehensive Cancer Network. PMID- 10370920 TI - Reproducibility of guidelines. A comparison of the NCCN and ASCO lung cancer guidelines. AB - Clinical practice guidelines are reproducible if two groups of experts, presented with the same evidence and methods, derive similar recommendations. Reproducibility is an essential attribute in justifying the use of clinical practice guidelines to guide and monitor physicians' practices. This article will compare guidelines for the management of advanced non-small-cell lung cancer developed by the National Comprehensive Cancer Network (NCCN) with guidelines derived by the American Society of Clinical Oncology (ASCO). This comparison aims to determine whether the two guidelines are similar or whether there are major differences that pose significant problems for clinicians attempting to manage patients according to one oncology standard. PMID- 10370922 TI - Outcomes assessment in the NCCN: 1998 update. National Comprehensive Cancer Network. AB - The assessment of outcomes is a major priority of the National Comprehensive Cancer Network (NCCN). To date, the NCCN's outcomes program has (1) made a systematic inventory of member institutions' existing data sources; (2) begun a collaborative project with the National Cancer Data Bank to compare specific surgical patterns of care in NCCN institutions with national norms; and (3) begun the creation of a prospective outcomes database within the NCCN. As the first step in the database initiative, five NCCN institutions participated in a pilot project in one disease, breast cancer, aimed at developing and testing techniques of data collection, aggregation, and analysis. Six more sites were added to the pilot program in 1998. As of September 1998, data on over 1,000 unique cases had been submitted to the database, with the number of additional cases expected to triple or quadruple within the ensuing 12 months. Future plans call for (1) further expansion of the breast cancer database, both in terms of the number of institutions participating and the scope of data being collected; (2) the addition of a second disease, non-Hodgkin's lymphoma, to the database; and (3) the establishment of partnerships with other organizations for whom the database might provide useful information, such as insurers, pharmaceutical and biotechnology firms, and regulatory and accrediting bodies. PMID- 10370923 TI - Financial modeling for global pricing: the NCCN breast cancer prototype. National Comprehensive Cancer Network. AB - In conjunction with its Outcomes Database, the National Comprehensive Cancer Network (NCCN) is developing a series of financial models to calculate the cost of various episodes of care for cancer. The prototype model is based on the NCCN Breast Cancer Practice Guidelines. This article describes how the model was developed, its constituent parts, and what information it is designed to provide. To accommodate the differences in costs in various areas of the United States, all costs and charges in the model are institution-specific. PMID- 10370924 TI - NCCN practice guidelines for Hodgkin's disease. National Comprehensive Cancer Network. PMID- 10370925 TI - NCCN practice guidelines for the management of psychosocial distress. National Comprehensive Cancer Network. PMID- 10370926 TI - NCCN colorectal cancer screening practice guidelines. National Comprehensive Cancer Network. PMID- 10370928 TI - Medicare, managed care, and cancer. AB - The Medicare program has undergone phenomenal changes during the last decade. Many of the changes relate to the growth of Medicare managed care. Changes in Medicare will continue as the Health Care Financing Administration implements the provisions of the Balanced Budget Act of 1997. As a result, Medicare should be open to more types of delivery systems that are organized in different ways than traditional health maintenance organizations. Payments should be improved so that these organizations can enroll sicker people and can be paid for accepting and treating those people. Beneficiaries should be provided with enough information that they have the necessary data to select plans based on quality, not just cost. PMID- 10370927 TI - Modulation of multidrug resistance: a paradigm for translational clinical research. AB - Resistance of cancer cells is the major limitation to the success of chemotherapy. Although many mechanisms of cellular resistance to anticancer drugs have been defined, the best understood of these is multidrug resistance (MDR), caused by the multidrug transporter, P-glycoprotein (P-gp), the product of the MDR1 gene. New drugs developed specifically to inhibit P-gp and modulate MDR, such as valspodar (PSC 833 [Amdray]), are currently undergoing clinical testing. Moreover, agents designed to inhibit other mechanisms of drug resistance are currently in development, and concurrent blockade of multiple mechanisms of resistance appears to be a promising approach. Coadministration of MDR1-related chemotherapeutic drugs with an MDR modulator may enhance the bioavailability of these agents sufficiently to enable oral dosing, which would potentially be more convenient and less toxic. PMID- 10370929 TI - NCCN practice guidelines for fever and neutropenia. National Comprehensive Cancer Network. PMID- 10370930 TI - Antidepressants in rehabilitation. AB - Antidepressant medications have a variety of uses in addition to the treatment of depression. This article focuses on the two most widely used categories of antidepressants, tricyclic antidepressants (TCAs) and the newer selective serotonin reuptake inhibitors (SSRIs), and highlights their use in post-stroke depression and chronic pain. PMID- 10370931 TI - Analgesics. Opioids, adjuvants, and others. AB - The purpose of this article is to summarize the main categories of pain-relieving medications. The authors review a number of analgesic preparations and treatments, with special emphasis on advantages, precautions, limitations, and various routes of administration. PMID- 10370932 TI - Antiepileptic drugs. AB - Antiepileptics are a very important class of medications, and the number of these drugs available for clinical use has increased dramatically in the last decade. The pharmacology and indications for use in a variety of physiatric patient groups are comprehensively and systematically reviewed. PMID- 10370933 TI - Anti-inflammatory therapy. AB - The inflammatory response involves a complex set of stereotyped biochemical and cellular reactions that aim to eliminate pathogens. Systemic inflammatory disorders, such as rheumatoid arthritis and systemic lupus erythematosus, feature a persistent, uncontrolled inflammatory response that typically culminates in tissue damage. The control of this inflammatory response is of paramount importance in preventing irreversible tissue damage, as well as in controlling symptoms. NSAIDs and corticosteroids are the most effective agents currently available to control the clinical manifestations of inflammation, although they usually need to be used in conjunction with other "disease modifying" strategies in order to achieve optimum control of chronic inflammatory disorders. As with all pharmacologic therapy, the risk to benefit ratio of each agent needs to be considered carefully before embarking on extended courses of therapy. Strategies to minimize the long-term risks of NSAIDs and corticosteroids are continuing to evolve and hold the promise of greater safety without loss of efficacy. PMID- 10370934 TI - Antihypertensives. AB - Management of hypertension requires a thorough knowledge of currently available antihypertensive drugs. This article reviews the mode of action of the major classes of antihypertensives, indications for their use, and adverse effects. The main emphasis is placed on their use, in physiatric practice. PMID- 10370935 TI - Antispasticity medications. AB - The article illustrates a practical approach to the challenging management of problematic, generalized spasticity. Use of dose titration to achieve symptomatic relief is described. Currently approved pharmaceuticals used as antispasticity agents and muscle relaxants and other medications with antispasticity effects are reviewed. PMID- 10370936 TI - Neuromuscular blockers. AB - The use of corrosive, injectable neuromuscular blockers has been a treatment option for many years; however, the more recent advent of botulinum toxin (BTX) treatment has revived interest in localized treatments. This article reviews the use of local anesthetics, alcohol, phenol, and BTX treatment for localized muscular overactivity syndromes. PMID- 10370937 TI - Intrathecal drug therapy. AB - The direct application of drugs for non-hospitalized patients became a practical therapeutic modality with the advent of implantable drug delivery devices, or "pumps". This article describes the use of pumps for the intrathecal infusion of baclofen, morphine and clonidine. PMID- 10370938 TI - Antimicrobial agents. AB - This article employs a comprehensive approach to review antimicrobial drugs, emphasizing the strengths and limitations of drugs including the traditional, those newly introduced and those currently under development. The significant issue of antimicrobial resistant organisms is addressed specifically. PMID- 10370939 TI - Medications used in the treatment of multiple sclerosis. AB - Recently, there have been numerous advances in the treatment of multiple sclerosis. This article focuses on reviewing the various forms of MS, managing its various symptoms, and possibly altering the course of the disease in an overall attempt to improve the quality of life of the patient. PMID- 10370940 TI - Neuroprotective agents. AB - Although basic research has revealed many mechanisms involved in the repair or elimination of damaged neurons, turning these mechanisms into clinically useful neuroprotective interventions is a slow process. Numerous neurotrophic factors seem to mediate neuronal repair and viability, but because the neurotrophic factors are proteins or polypeptides, they cannot be given orally and do not enter the brain if given intravenously. Tapping into the neuroprotective potential of the neurotrophic factor mechanisms must await further developments. Similarly, pharmacological agents that protect damaged neurons by reducing glutamate excitotoxicity, by scavenging free radicals, or by increasing adenosine inhibitory influences, are not ready yet for widespread clinical use. Also, appropriate therapeutic protocols for currently available neuroprotective agents such as vitamin E, selegiline, and NSAIDs remain to be determined. Given the rate of advance of research in this area, however, meaningful neuroprotection and neurorescue will be attainable in the very near future. In the meantime, neuron damaging oxidative stress can be kept in check by insuring adequate dietary sources of antioxidants. Although there is as yet little or no scientific evidence that dietary antioxidants are neuroprotective, the consumption of high antioxidant foods, such as blueberries and strawberries, is appealing to most people regardless of any neuroprotective potential. PMID- 10370941 TI - Tranquilizers, stimulants, and enhancers of cognition. AB - For patients with cognitive dysfunction, the appropriate use of neurotransmitter specific medication may permit independent living. This article examines a variety of tranquilizers, stimulants, and enhancers of cognition and evaluates which may be most effective given their side effect profiles. PMID- 10370942 TI - Urinary and gastrointestinal systems medications. AB - This article reviews the medical management of the neurogenic bladder and bowel. The drugs discussed specifically affect detrusor instability, detrusor weakness, high urethral pressure, low urethral closure pressure, inflammatory cystitis, and chronic constipation. PMID- 10370943 TI - Special considerations for medication use in children with developmental disabilities. AB - Regardless of the underlying cause of a child's developmental disability, a similar constellation of associated medical conditions may occur. Successful management approaches in children with developmental disabilities are generally interdisciplinary, frequently involving a variety of medical and surgical disciplines, allied health professionals, and the child's family. Pharmacologic intervention is an integral aspect of the management plan, but consideration must be given both to the possible effects of the medications on a child's growth and development, and to the avoidance of adverse drug interactions and reactions. PMID- 10370945 TI - [Place of surveillance in the treatment of localized prostate cancer]. AB - Surveillance with deferred treatment in patients with clinically prostate confined prostate cancer is a controversial subject. The policy of prostate cancer screening for increasingly low PSA levels and the use of more extensive biopsy protocols result in the detection of a large number of small tumours, probably early in their subclinical course. Conversely, some patients who receive initial active treatment are understaged by the pretreatment assessment and develop treatment-related adverse effects without any oncological benefit. The main published series concerning surveillance were analysed with particular emphasis on those with the longest observation periods. Data concerning specific survival and progression-free survival are discussed, together with the results obtained in terms of the quality of life of patients submitted to surveillance. Analysis of these data show a potential benefit of surveillance in elderly patients (over the age of 70 years) with small, low-grade prostatic tumours. PMID- 10370944 TI - [Place of chemotherapy in the treatment of invasive bladder tumors]. AB - Urothelial bladder tumours are chemosensitive. Chemotherapy is indicated in the case of metastatic bladder cancer. The M-VAC protocol remains the reference treatment. The efficacy of this protocol is estimated to be about 18% in terms of complete responses and 20% of these responding patients achieve long-term survival. New combinations, comprising drugs such as ifosfamide, gallium nitrate, paclitaxel or gemcitabine, appear to be promising. Neoadjuvant and adjuvant chemotherapy cannot be considered to be standard treatment at the present time. PMID- 10370946 TI - [Comparison of spiral computed tomography without contrast media and intravenous urography in the diagnosis of renal colic]. AB - OBJECTIVES: This prospective study was designed to determine the reliability of noncontrast spiral CT scan, compared to that of intravenous urography (IVU), in the diagnostic assessment of acute renal colic. MATERIAL AND METHODS: 53 patients, admitted with an empirical diagnosis of renal colic after initial assessment, were included and underwent spiral CT scan without contrast agent injection, immediately followed by IVU. These examinations were performed according to a blind protocol by two different radiologists. We initially looked for the presence of stones and/or urinary tract obstruction. Patients with a stone and/or obstruction (urinary tract dilatation) visible on CT scan were then examined for the presence of CT signs associated with stone disease (infiltration of the perirenal and periureteric fat, oedema of the ureteric wall and loss of sinus fat). We calculated the statistical correlation between the presence or absence of obstruction and these accessory signs. RESULTS: 45 stones were recovered (in 36 cases before the two examinations). 36 stones were identified on CT versus only 24 on urography. Urinary tract dilatation was demonstrated in 26 out of 53 cases by both urography and CT. The frequency of accessory signs visible on CT in the presence of stones (n = 36) was 66% for infiltration of periureteric fat, 36% for infiltration of perirenal fat and 75% for oedema of the ureteric wall. In the presence of urinary tract dilatation, these frequencies were 92%, 84% and 60% respectively. CONCLUSION: Noncontrast spiral CT is a reliable and rapid diagnostic modality for the detection of urinary stones, providing a morphological study equivalent to that of IVU and able to guide appropriate treatment. It should replace IVU in the diagnostic assessment of renal colic. PMID- 10370947 TI - [Renal retransplantation in adults. Comparative prognostic study]. AB - For a long time, retransplanted patients were considered to be high-risk subjects, but a number of studies have shown that retransplantation can give good short-term results. OBJECTIVES: Between October 1987 and February 1997, 51 retransplanted patients (Group 1) were compared with 96 patients (Group 2), matched for age, sex and date of transplantation, receiving a first kidney, with a mean age of 41 +/- 10 years and a mean follow-up of 44 +/- 32 months. RESULTS: The patient did not differ in terms of aetiology of the renal disease, mismatches and duration of dialysis before the first transplant and the duration of dialysis before the second transplant was 53 +/- 54 months. In Group 1, transplant loss was due to an immunological [34], surgical [8], or medical [3] cause, or due to recurrence of the disease [3] and discontinuation of treatment [5]. Hyperimmunized subjects were more numerous in Group 1: 21.4% vs 5.2% (p < 0.01). Immunosuppression only differed in terms of Cyclosporin: 65% for Group 1 vs 45% for Group 2 (p < 0.05). The two groups did not differ in terms of acute rejection (33% for Group 1 vs 48% for Group 2), serum creatinine at last review (140 +/- 51 +/- 65 m mol/l) and 5-year and 8-year patient (92 vs 82% and 92 vs 76%) and graft survival curves (85 vs 75% and 59 vs 61%). CONCLUSION: This study confirms that retransplantation can be successfully performed, even in terms of long-term results. PMID- 10370948 TI - [Treatment of ureteropelvic junction stenosis with cold blade retrograde endopyelotomy]. AB - BACKGROUND: The classical treatment of ureteropelvic junction (UPJ) stenosis consists of open surgical resection-anastomosis. Endopyelotomy, a less invasive procedure, allows antegrade or retrograde endoscopic incision of this junction. The authors' experience of cold blade retrograde endopyelotomy (CBRE) is presented and compared with other techniques. MATERIAL AND METHOD: Thirteen patients (3 men and 10 women), with a mean age 55.3 years, underwent CBRE with ureteroscopic and fluoroscopic control for primary (10/13) or secondary (3/13) UPJ stenosis. The clinical and urographic efficacy of the method was assessed retrospectively. RESULTS: Median operating times and hospital stays were 40 min and 4 days respectively. The postoperative complication rate was 15.4% (two urinary tract infections). Eleven of the 13 patients are currently symptom-free with a mean follow-up of 26.5 months. Follow-up urography (mean follow-up: 11.9 months) shows a satisfactorily revascularized junction in 84.6% of cases (1 asymptomatic stenosis, 1 symptomatic stenosis revised by open surgery). CONCLUSION: Compared to the other treatments of UPJ stenoses, CBRE is a simple, minimally invasive technique, which reduces operating time and hospital stay and therefore presents a good cost-efficacy ratio. PMID- 10370949 TI - [Primary small cell carcinoma of the bladder]. AB - OBJECTIVES: Primary small cell carcinomas of the bladder differ from transitional cell carcinomas by their rarity, histological characteristics, malignant potential and treatment. This study analysed the diagnostic criteria and therapeutic results obtained in a consecutive patient series over a 6-year period. MATERIALS AND METHODS: 7 patients (6 men and one woman) suffering from primary small cell carcinoma of the bladder were evaluated. Histological slides, treatment modalities and duration of survival were reviewed. RESULTS: The commonest clinical presentation was macroscopic haematuria. All tumours were invasive at the time of diagnosis. Two patients were treated by partial cystectomy, one of whom also received adjuvant chemotherapy. One patient was treated by radical cystectomy and 4 also received adjuvant chemotherapy, including 2 with neoadjuvant radiotherapy at a dosage of 65 Gy. The three patients treated by a single treatment modality (surgery alone or chemotherapy alone) had a shorter survival, in contrast with patients treated by a combination of chemotherapy and/or surgery. CONCLUSION: Primary small cell carcinomas of the bladder are rare and have a poor prognosis. Treatment must consist of a combination of neoadjuvant or adjuvant chemotherapy and surgery or radiotherapy to achieve the best results. PMID- 10370950 TI - [Prevention of temporary prostatic obstruction after high-energy microwave thermothery by placement of prostatic bridge catheter]. AB - OBJECTIVES: The clinical utility of a novel intraurethral prostatic bridge catheter (PBC) was evaluated for prevention of temporary prostate obstruction following targeted high-energy transurethral microwave thermotherapy (TUMT) in patients with benign prostatic hyperplasia (BPH). MATERIAL AND METHODS: High energy TUMT was administered to 54 BPH patients under topical urethral anesthesia followed by placement of a PBC, which remained indwelling up to 1 month. Patient evaluation included determination of peak urinary flow rate (Qmax), International Prostate Symptom Score (IPSS), and quality of life (QOL) score at baseline, immediately following TUMT and PBC placement, and at periodic intervals thereafter up to 1 month. Results were compared retrospectively with those of 51 patients who underwent TUMT followed by standard temporary urinary catheterization, generally for 24 h. RESULTS: Immediately following TUMT and PBC placement significant improvements (p < 0.0005) were observed in mean Qmax, IPSS and QOL score of 59.3%, 33.5% and 23.6% respectively, compared with baseline values. Further improvements were demonstrable up to 1 month, at which time mean Qmax, IPSS and QOL score had improved 79.0%, 54.9% and 56.5%, respectively, vs baseline means (p < 0.0005). In a retrospective comparison at baseline and 14 days between PBC recipients (PBC group) and a cohort of TUMT patients who had undergone temporary standard catheterization and subsequent catheter removal (standard catheterization group), mean baseline Qmax, IPSS and QOL score were similar between the two groups. However, at the 14 day follow-up evaluation in the PBC group mean Qmax was 101.8% higher, and IPSS and QOL score were 47.9% and 51.1% lower, respectively, than the corresponding values in the standard catheterization group (p 0.0005). The PBC was well tolerated and remained in situ throughout the entire 1 month follow-up period in 48/54 (88.9%) patients. Early PBC removal was performed in 3/54 patients (5.6%) because of urinary retention and in 3/54 patients (5.6%) due to PBC migration. During the acute post-TUMT recovery period, PBC recipients experienced impairment in sexual function which, though statistically significant, was comparatively small in magnitude. CONCLUSION: PBC provides an efficacious and well-tolerated option for preventing prostatic obstruction in the acute post-TUMT period. This approach avoids the inconvenience and infection risk of standard indwelling catheters or intermittent self-catheterization. PBC insertion and removal are rapid, facile and non traumatic. PBC placement may prove useful in improving the early results of TUMT. PMID- 10370951 TI - [Prospective follow-up of 3,228 patients suffering from clinical benign prostatic hyperplasia (BPH) treated for 3 years wi alfuzosin in general practice. BPH Group in General Practice]. AB - OBJECTIVE: To determine (a) the amplitude and duration of reduction of the symptom score and improvement of the HRQL score (including sexual function), (b) the adverse effects and (c) the incidence of acute urinary retention and prostatic surgery during the 3 years of alfuzosin treatment. MATERIAL AND METHODS: 3,228 patients suffering from BPH were included by 812 centers in a 3 year open prospective study and were treated with alfuzosin (immediate release) at the recommended dosage. A symptom score (modified Boyarsky) and a specific HRQL score, comprising 20 items including 3 questions on sexuality (Urolifetm BPH Qol20) were self-administered on inclusion and after 3, 6, 12, 18, 24, 30 and 36 months. RESULTS: 2,579 patients (79.9%) completed the 3 years of the study. The symptom score was significantly decreased by 54% at 3 months and this reduction was maintained until 36 months (-48.4%); the HRQL score was significantly improved by 45.4% at 12 months and this improvement was maintained until 36 months (+43.4%). Alfuzosin was well tolerated: the qualitative and quantitative distribution of adverse effects was identical to that previously observed in placebo-controlled trials (vertigo-dizziness: 2.1%). Adverse effects were responsible for 4.2% of drop-outs from the trial. 120 patients (3.7%) were operated for BPH and 9 patients (0.3%) developed acute urinary retention. CONCLUSION: This prospective study confirms the long-term safety of use of alfuzosin under routine general practice conditions and emphasizes the need to measure HRQL in the context of the patient's opinion. PMID- 10370952 TI - [Value of free PSA/total PSA ratio in therapeutic decisions in the case of a single positive biopsy of the prostate]. AB - OBJECTIVES: Study of the value of the free PSA/total PSA ratio in the therapeutic decision concerning prostatic adenocarcinoma, in the case of a single positive biopsy. MATERIAL AND METHODS: The free PSA/total PAS ratio was calculated on serum samples derived from 37 patients with clinically localized prostatic carcinoma and only one positive biopsy, in whom radical prostatectomy was performed. RESULTS: The free PSA/total PSA ratio appeared to be independent of pathological stage and histological prognostic criteria (grade and score, degree of capsular effraction). CONCLUSION: In the case of a single positive biopsy, calculation of the free PSA/total PSA ratio does not appear to provide any decisional criteria in favour of radical prostatectomy. PMID- 10370953 TI - [Can preoperative urodynamic examination allow us to predict the risk of incontinence after radical prostatectomy?]. AB - OBJECTIVES: To verify whether bladder dysfunction detected by urodynamic studies prior to radical prostatectomy can predict postoperative continence status. MATERIAL AND METHODS: Twenty patients diagnosed with prostate cancer had multichannel subtracted filling and voiding videocystometry before undergoing radical retropubic prostatectomy. Postoperatively, all patients had periodic clinical assessment of continence status. RESULTS: On preoperative filling cystometry, detrusor instability with a maximal detrusor pressure greater than 15 cm H2O was demonstrated in 12/20 patients (60%). Postoperatively, 11/20 patients (55%) were continent, 4 (20%) had mild stress incontinence and 5 (25%) complained of episodic urge incontinence. However, only 5 of the 12 patients with preoperatively diagnosed detrusor instability manifested clinical urge incontinence after surgery (positive predictive value = 41.6%). CONCLUSION: The incidence of preoperative detrusor instability in our series was high, but little correlation was found between this finding and postoperative incontinence. PMID- 10370954 TI - [Cancer of the urethra in women: experience of the National Institute of Cancer of Brazil: 1992-1997]. AB - OBJECTIVE: Describe the results of surgery, radiation therapy and chemotherapy for treatment of primary malignant neoplasms of female urethra. PATIENTS AND METHODS: Since 1982, 31 patients with urethral cancer were evaluated at our institution (follow-up ranging from 0 to 127 months). Ten patients were treated with external beam irradiation and 1 patient received preoperative therapy after surgery. Two patients refused treatment and 1 received chemotherapy. Three patients presented with disseminated metastatic disease at the first examination received exclusively palliative treatment. RESULTS: Five patients have survived from 3 to 10 years following treatment without recurrence. Of 31 patients 18 developed distant metastases during the first 24 months of follow-up irrespective of treatment employed. CONCLUSION: With exception of primary melanoma, prognosis was not related to histologic features. Patients who underwent surgery and radiotherapy had a better survival rate than did those who received radiotherapy alone. Total urethrectomy with appendico-vesicostomy can be an alternative surgical method for entire urethral ivnasive lesions without cystourethrectomy and preserving urinary continence. PMID- 10370955 TI - [Leydig cell testicular tumors. A series of 10 observations]. AB - OBJECTIVES: To define the clinical and laboratory characteristics and natural history of Leydig cell tumours in order to define a general management plan. MATERIAL AND METHODS: The files of 10 patients operated for Leydig cell testicular tumour between 1982 and 1996 were studied retrospectively. RESULTS: In nine out of ten cases, the presenting complaint was gynaecomastia, erectile dysfunction or infertility. In every case, serum testosterone was normal or low and oestradiol was normal or elevated. Eight patients were treated by radical orchidectomy, and two by subcapsular orchidectomy. The course was favourable in 9 out of 10 cases in the absence of any other treatment. Only one patient had an immediately malignant form with a fatal outcome. CONCLUSION: Although many teams prefer total orchidectomy because of the diagnostic difficulty associated with malignant forms, simple subcapsular orchidectomy should become the first-line treatment, provided it is subsequently followed by close surveillance, as it preserves maximum fertility, and these tumours usually have a favourable prognosis. PMID- 10370956 TI - [Human ejaculation: physiology, surgical conservation of ejaculation]. AB - OBJECTIVE: The most widely accepted theory to explain ejaculation consists of two phases: 1) A phase of accumulation of the various constituents of semen inside the prostatic urethra. 2) A phase of expulsion with opening of the striated sphincter, while the smooth sphincter of the bladder neck remains closed. The objective of this study was therefore to confirm or invalidate this hypothesis. MATERIAL AND METHODS: Eight volunteers were studied by continuous transrectal ultrasonography allowing dynamic analysis of ejaculation. Sixteen patients presenting with a bladder neck obstruction syndrome were treated by a technique preserving the last centimetre of supramontanal urethra. RESULTS: In the eight healthy subjects, we constantly and successively observed: filling of the ejaculatory ducts. prostatic contractions. Expulsion of the contents of the seminal vesicles into the inframontanal urethra, without prior ballooning of the prostatic urethra. In the 16 patients treated for bladder neck obstruction, we did not observe any case of retrograde ejaculation. PMID- 10370957 TI - [Primary leiomyosarcoma of the kidney. A case report]. AB - Primary sarcomas of the kidney are exceptional, representing 1 to 3% of all renal tumours in adults. They have a poor prognosis. The authors report a case of primary leiomyosarcoma of the kidney in a 44-year old patient, presenting in the form of low back pain and haematuria. Treatment consisted of radical nephrectomy. Two months later, the patient presented with hepatic metastases. The patient is currently receiving chemotherapy. PMID- 10370959 TI - [Retroperitoneal mucinous cystadenoma]. AB - The authors report an unusual case of retroperitoneal mucinous cystadenoma in contact with the isthmus of a horseshoe kidney and associated with congenital absence of the left ovary. The retroperitoneal site is very unusual, as only about twenty cases have been reported in this site. The pathogenesis of these tumours remains unclear and the authors discuss the various hypotheses proposed to date. PMID- 10370958 TI - [Puerperal thrombophlebitis of the ovarian vein revealed by renal colic]. AB - The authors report the clinical case of a young woman with thrombophlebitis of the right ovarian vein following delivery by caesarean section, initially presenting in the form of renal colic. In the light of a review of the literature, they recall the pathophysiological mechanisms of ovarian thrombophlebitis and the various features observed on imaging examinations. The most frequent clinical features are also described. The authors emphasize the potential, but rare severity of this disease, characterized by the risk of pulmonary embolism, and its treatment, which is usually medical. PMID- 10370960 TI - [Traumatic dislocation of the testis. Report of three cases]. AB - The authors report three cases of traumatic dislocation of the testis. Each case concerned a young patient, victim of a motorbike accident with direct trauma to the perineum and scrotum on the reservoir. Preoperative radiological assessment consisted of ultrasound, computed tomography, or even magnetic resonance imaging, depending on the case. Surgical exploration revealed 3 traumatic dislocations of the testis, including one with associated torsion of the spermatic cord. One case also presented fracture of the contralateral testis, which was not dislocated. Each case was repaired by repositioning of the testis in the scrotum. The fractured testis could not be preserved. A review of the literature confirmed the rarity of these traumatic lesions. Dislocations are usually inguinal, sometimes bilateral, and can be ectopic. Spermatic cord lesions may also be associated. The authors define the place of radiological assessment, which can guide the diagnosis and therapeutic strategy. Surgical exploration, allowing assessment of the lesions, is required in every case. Repositioning of the testis in the scrotum ensures cure when the testis is still viable. PMID- 10370961 TI - [Perineal lipoma and accessory scrotum]. AB - An accessory scrotum associated with perineal lipoma is extremely rare. We report a case of a median lipoma of the perineum in an accessory scrotum. The literature is reviewed. PMID- 10370962 TI - [Risk of accidental contamination by the human immunodeficiency virus (HIV): review and management]. AB - In surgery practice, everyone must know what to do in case of injury with exposure to body fluids. The chimioprophylactic treatment has to be easy to take in case of high risk of VIH transmission. To prevent these accident, surgery on infected people must always be done by surgeon who have experience. PMID- 10370963 TI - [Pressure-flow urologic analysis in children: reflections after 20 years of practice]. AB - Pressure-flow analysis in children allows optimal definition of the main urodynamic parameters of micturition and clarifies certain aspects of physiology and functional disorders. This examination has allowed urologists to establish reliable relationships between the findings of the clinical interview and clinical examination and the reality of urodynamic parameters, so that, after being widely used for more than 20 years, its indications have now become exceptional, limited to the detection of detrusor-sphincter pathophysiology and a few rare voiding disorders, poorly defined by clinical findings or imaging. PMID- 10370964 TI - [Pressure-flow analysis of micturition in men in clinical practice]. PMID- 10370965 TI - [Pressure-flow analysis of micturition in women in clinical practice]. PMID- 10370966 TI - [Mathematical model of micturition allowing a detailed analysis of free urine flowmetry]. AB - A mathematical model of micturition allowing precise analysis of uroflowmetry curves (VBN method) is described together with some of its applications. The physiology of micturition and possible diagnostic hypotheses able to explain the shape of the uroflowmetry curve can be expressed by a series of differential equations. Integration of the system allows the validity of these hypotheses to be tested by simulation. A theoretical uroflowmetry is calculated in less than 1 second and analysis of a dysuric uroflowmetry takes about 5 minutes. The efficacy of the model is due to its rapidity and the precision of the comparisons between measured and predicted values. The method has been applied to almost one thousand curves. The uroflowmetries of normal subjects are restored without adjustment with a quadratic error of less than 1%, while those of dysuric patients require identification of one or two adaptive parameters characteristic of the underlying disease. These parameters remain constant during the same session, but vary with the disease and/or the treatment. This model could become a tool for noninvasive urodynamic studies. PMID- 10370967 TI - [Obstructive effect of a urethral catheter of urination parameters: theoretical study]. PMID- 10370968 TI - [Penile disassembly technique: a new approach in the surgical reconstruction of hypospadias]. AB - We report our experiences with penile disassembly technique in the treatment of severe hypospadias. During a period from November 1995 to December 1997 the technique was applied to 102 patients, aged from 9 months to 32 years. The principle of the technique involves separation of the penis into its component parts: glans cap with neurovascular bundle (dorsally) together with undivided or divided urethra and urethral plate (ventrally), and corpora cavernosa. After correction of the curvature and different techniques of urethroplasty the penile entities are joined into normal anatomical relationships. Our goal was also to achieve some degree of penile enlargement in small hypospadiac penises. The patients were followed from 4 to 29 months (mean 19.2 months). Straightening of the penis was achieved in all cases without recurrence of curvature. Complications were related to urethroplasty: 3 urethral stenoses, 2 fistulas and 2 diverticulum. Penile disassembly technique is very effective for most severe hypospadias. Possibility of penile augmentation is real with this technique. PMID- 10370969 TI - The contribution of molecular genetics in the diagnosis and management of neuromuscular disorders. AB - It is true that the recent advances in molecular genetics have generated a medical revolution. This is especially true for the inherited neuromuscular disorders. There have been many spectacular recent discoveries with new genes being found and their protein products identified. One of the most remarkable aspects of this progress is the nexus which has developed between the basic discovery and its clinical application. As soon as a new genetic mutation is reported, the information may be used immediately to establish the molecular diagnosis for that disorder in any part of the world which has a DNA laboratory. This is done by using primers derived from the published DNA sequences using the polymerase chain reaction (PCR). This development is of immense value for the clinician as it provides an exact molecular diagnosis often with prognostic information and the test results can be used for genetic counselling and prenatal diagnosis. One of the unexpected outcomes of this work has been the surprising variation which has been shown to exist between genotype and phenotype. Previously, one mutation was believed to be responsible for one clinical disorder. However, it is now known that one genotype may be responsible for a variety of phenotypes and vice versa. In the field of neuromuscular disorders the most notable advances have occurred for Duchenne muscular dystrophy and the related dystrophinopathies and for the group of limb girdle muscular dystrophies, especially the subgroup of sarcoglycanopathies. Other areas are the congenital myopathies, the 'channel-opathies' and the mitochondrial cytopathies. In this review the most commonly used molecular genetic and immunocytochemical methods using antibodies to the protein product are outlined together with the principles of their application in the neuromuscular clinic. Included are the provisos and pitfalls which need to be kept in mind in the interpretation of DNA results for each patient. PMID- 10370970 TI - Problems and solutions in the rehabilitation of patients with progressive muscular dystrophy. PMID- 10370971 TI - Muscular dystrophy: genetic counselling and family planning. PMID- 10370972 TI - Mitochondrial myopathies. Clinical and diagnostic aspects. PMID- 10370973 TI - Dystrophia myotonica. Clinical, pathophysiological and molecular aspects. PMID- 10370974 TI - Congenital muscular dystrophy. Care of children and families. PMID- 10370975 TI - Assessment of motor functions. PMID- 10370976 TI - Physiotherapy in muscular dystrophy. PMID- 10370977 TI - The impact on British blood services on BSE and v-CJD: implications for patients, donors and public health. PMID- 10370978 TI - The role of leukotriene receptor antagonists (LTRAs) in the management of asthma. PMID- 10370979 TI - Treatment for carotid atherosclerosis--who should have it and who should not? PMID- 10370980 TI - Seasonality of presentation of type I diabetes mellitus in children. Scottish Study Group for the Care of Young Diabetics. AB - Environmental influences are thought to have an aetiological role in onset of diabetes in children. Month of onset in over 2000 children in Scotland was established and there was an excess in colder/darker months than in warmer/lighter months. A meta-analysis of 21 previous studies with over 13,000 patients gave the same result at a much higher level of significance. A mechanism is postulated based on previous viral induced islet cell damage with ongoing progressive auto-immune destruction. There may be physiological seasonal changes with winter stress on carbohydrate and lipid metabolism. The raised winter levels of pituitary, adrenal and thyroid hormones fail to be antagonised by falling level of insulin. A role for seasonal variation in exercise and nutrition is considered. PMID- 10370981 TI - Treatment limiting decisions in a neurosurgical unit. AB - In a previous paper on mortality audit we reported on the use of treatment limiting decisions (TLDs) in a neurosurgical unit in the year 1988. In this paper we compare the findings of a similar audit for 1997. It appears that our unit's policy of openly discussing all TLDs in patients who die had led to such decisions being made at a more appropriate stage in the patient's illness. Regular review of TLDs is probably helpful in increasing the confidence of clinicians to make these difficult decisions openly and timeously. PMID- 10370982 TI - Changes in skeletal and cardiac muscle enzymes during the Scottish Coast to Coast Triathlon. AB - While skeletal muscle injury is common after prolonged exercise, evidence in the literature supporting cardiac muscle injury is conflicting. Creatine kinase and cardiac troponin-I were measured, in 31 amateur athletes (25 male) before, and 12 24 hours after, a 300 km cycling/running/canoe triathlon event. A short questionnaire was used to assess level of fitness, training and previous experience. Creatine kinase levels were greater after the 45 km cross-country run compared with after a 155 km road cycle (60.5 +/- 62.8 iu/L/kg vs 19.3 +/- 9.6 iu/kg, P = 0.03). Individuals performing running and cycling events consecutively had creatine kinase similar to those observed after running alone (50.2 +/- 53.8 iu/L/kg vs 60.5 +/- 62.8 iu/L/kg, P = 0.55). Cardiac troponin-I was elevated above the normal range (0.1 ng/L) in six athletes (four in running and cycling events, one in the running and one in the cycling event). We conclude that running produces significantly more skeletal muscle injury than cycling and that strenuous endurance exercise involving running and cycling in amateur trained athletes is associated with release of cardiac specific enzymes. The functional and longer term consequences of this require further study. PMID- 10370983 TI - The influence of a nurse practitioner on out of hours work intensity for surgical house officers. AB - We have prospectively studied the influence of a nurse practitioner service on out of hours work intensity of surgical house officers. Data collection was achieved by prospective audit. The study was set in the surgical wards in a large teaching hospital. The main outcome measures were; 1) the nature and frequency of overnight calls to the nurse practitioner, and 2) the outcome of these calls (doctor not bleeped, telephone advice given by doctor or doctor attended ward). A total of 645 calls were made over the 75 night study period (8.6 calls/night). Two hundred and ninety-six calls were managed by the nurse practitioner alone. This represents a 46% reduction in work intensity for the surgical house officer. This study illustrates the benefits of a nurse practitioner service and also identifies important areas for undergraduate education in preparing medical students for the common problems encountered during the surgical on-call period. PMID- 10370984 TI - Antiplatelet therapy for secondary stroke prevention. AB - The precise role of antiplatelet therapy in secondary stroke prevention remains a matter of some debate. Although specific antiplatelet agents (notably aspirin, ticlopidine, dipyridamole, and clopidogrel) have been shown to be active in the prevention of secondary stroke, questions remain about the effective dose of these agents and their potential efficacy in combined therapeutic regimens. In addition, haematological and gastrointestinal side-effects of antiplatelet agents remain a significant clinical concern for patients and prescribing physicians. This review article examines research on both monotherapy and combination antiplatelet therapy for secondary stroke prevention, with an emphasis on lessons learned about dosage schedules, treatment protocols, and side-effect profiles. PMID- 10370985 TI - Audit of the effect of introducing local guidelines for referral for carotid duplex scanning. AB - Guidelines for referral of patients for carotid duplex scanning were introduced stating that scans were only indicated for patients with hemispheric localising symptoms or asymptomatic bruits. A 19% reduction in referral rate was seen for new scans in a one year period. The number of "inappropriate" requests fell from 27.6% in 1994 to 12.8% in 1995. There was no reduction in the numbers of patients identified with effectively occult asymptomatic disease who may have been suitable for surgery in the same period. Guideline introduction had the effect of reducing unnecessary scans without any effect on the overall pick-up, rate for carotid disease. PMID- 10370986 TI - Anatomic basis of chronic groin pain with special reference to sports hernia. AB - Chronic pain on the ventral surface of the scrotum and the proximal ventro-medial surface of the thigh especially in athletes has been diagnosed in various ways; recently, in Europe the concept of "sports hernia" has been advocated. However, since few reports discuss the detailed course of the nerves in association with the pain, we examined the cutaneous branches in the inguinal region in 54 halves of 27 adult male cadavers. From our results, in addition to the cutaneous branches from the ilioinguinal n. (in 49 of 54: 90.7%), cutaneous branches originating from the genital branches of the genitofemoral nerve were found in the inguinal region in 19 of 54 halves (35.2%). In 7 cases (in 7 of 54: 13.0%) the genital branch and the ilioinguinal nerve united in the inguinal canal. In 6 cases the genital branch pierced the inguinal lig. to enter the inguinal canal, and in three cases the genital branch pierced the border between the ligament and the aponeurosis of the obliquus externus m. to be distributed to the inguinal region. Therefore, the courses of the genital branches vary considerably, and may have a very important role in chronic groin pain produced by groin hernia. In addition, entrapment by the ligament may be a reasonable candidate for the cause of chronic groin pain. PMID- 10370988 TI - Anatomy of the common trunk of the middle and left hepatic veins: application to liver transplantation. AB - An anastomosis between the common trunk of the middle and left hepatic veins of the receiver and the cranial portion of the inferior vena cava of the donor is one of the techniques for restoration of hepato-caval continuity in orthotopic liver transplantation. This technique avoids dissection of the retrohepatic vena cava and total caval clamping. The aim of this study was to define the feasibility of this technique by a morphologic and biometric study of the common trunk of the middle and left hepatic veins on the basis of 64 injection-corrosion hepatic specimens and 21 fresh subjects. A common trunk for the middle and left hepatic veins was present in 54 of 64 cases (84%) with a length of 3 to 17 mm. The diameter of the new ostium constructed by section 0.5 cm proximal to the junction of the middle and left hepatic veins was 23.9 +/- 2.3 mm, which approximated to that of the vena cava where it traversed the diaphragm (24.4 +/- 2.0 mm). These findings confirmed that restoration of hepato-caval continuity by anastomosis between the common trunk of the middle and left hepatic veins of the receiver and the cranial portion of the vena cava of the graft is possible without incongruence. This study makes no assumptions about the hemodynamic effects associated with the smallest diameter of the true ostium of the common trunk at its opening into the inferior vena cava. In this study, the morphology of the common trunk was comparable to that observed by Nakamura. Further, we propose an anatomo-clinical classification allowing evaluation of the facility of vascular control of the common trunk in terms of the number and location of the collateral veins. PMID- 10370987 TI - Anatomic basis of minimal anterior extraperitoneal approach to the lumbar spine. AB - Anterior lumbar spine approaches may be indicated for fusion in degenerative lumbar spine disorders or to fill discal and bone gaps after fracture reduction. We present an anterior extraperitoneal approach applicable to any discal and vertebral levels from T12 to S1. The anatomic study, based on 25 cadavers, highlights retroperitoneal dissection principles for easy kidney and duodenopancreatic mobilisation and direct left anterior access to the entire lumbar spine. We established a precise description of the lumbar veins and the anastomoses between the left renal vein and hemiazygos system, in order to define different topographic and anatomic factors related to safe and easily reproducible approaches for cage or graft implementation. Independent of the level and previous intraperitoneal surgery, lumbar spine access with this approach safeguards the kidney, ureter, spleen, hypogastric plexus and duodenopancreatic system. Regarding operating time, blood-loss and possibilities for freshening and grafting, this technique seems an effective counterbalance to the difficulties and complex technology of endoscopic approaches. The clinical study includes our first 42 cases in traumatic and degenerative lesions. Avoiding the neurologic or hemorrhagic risk inherent in classical posterior lumbar interbody fusion (PLIF) techniques, it can be considered as a reasonable and valid alternative. This technique could be used in the near future for mini invasive discal prosthesis insertion. PMID- 10370990 TI - The saphenous venous compartments. AB - The relationships between the connective framework of the lower extremity hypodermis and the saphenous veins was studied by dissection, stereomicroscopy, ultrasonography and histology in 64 lower limbs. A fibroelastic lamina was evidenced in the hypodermis of the medial aspect of the thigh and leg and in the back face of the leg. This lamina, together with the underlying muscular fascia, fixed the boundaries of two compartments occupied by the saphenous veins and nerves. The adventitia of the saphenous veins was connected to the compartment walls by thick connective strands. The saphenous veins ran deeply in the hypodermis, closely ensheathed by a fibroelastic sleeve. As a consequence, they could no longer be considered as a truly superficial vein. This term seems to be appropriate only for their tributaries, which ran in a more superficial plane just below the dermis. The role of the saphenous vessels in blood return from the lower limbs may be greater than classically accepted. In fact, due to their close fascial ensheathing and adventitial anchoring, muscular contractions may enhance blood flow within these vessels as occurs in the intermuscular veins. Finally, dilative pathology of the saphenous vein may be resisted by the membranous lamina as a sort of a fibroelastic shield. PMID- 10370989 TI - Limb immobilization and intimal hyperplasia--an echo-Doppler study in man. AB - The authors studied the circulatory alterations observed in the upper limbs of patients showing muscular atrophy due to flaccid paralysis of traumatic origin. Ten patients who had suffered avulsion of the nerve roots from C4 to D1 that occurred 9 to 216 months previously, presented a significant degree of muscular atrophy of the affected upper limb, despite physiotherapy. We performed echo doppler examinations of the patients to measure the lumen of the subclavian, axillary, brachial, radial and ulnar arteries and also of the veins of both upper limbs. The measurements from both upper limbs in each patient were statistically compared. The data revealed a significant reduction of the width of the lumen of the arteries and veins and a reduced arterial blood flow in the affected limb in comparison with the normal one. The greater echogenicity and the abnormal Doppler waves of the affected vessels suggest that they presented an increased thickness and a hardened wall. The authors propose that this finding may be related to an intimal hyperplasia brought about by muscular atrophy or by the observed blood flow reduction. PMID- 10370991 TI - Elastic reinforcement and thickness of the joint capsules of the lower cervical spine. AB - Mechanical studies have shown the major strength of the joint capsules of the lower cervical spine, especially in its ventrolateral part. The aim of this study was to examine the structure of the joint capsules in order to discover if there is a correspondence between biomechanical properties and descriptive anatomy. Ten transverse sections and 4 sagittal sections obtained from 6 cadavers were observed under light microscopy at X 25 to X 250 magnification. Standard stains and specific elastic fiber stain were used for histologic preparation. The data were the thickness of the joint capsules in the different quadrants, and the topography and direction of the elastic fibers. The results showed that the ventrolateral part of the joint capsules is thick and reinforced by oblique elastic fibers. The dorsal part is thin. The authors suggest that the descriptive anatomy of the joint capsules confirms their mechanical properties. They note that the role of the ventrolateral part is supplemented by that of the posterior longitudinal ligament for the stability of the functional cervical spinal unit. PMID- 10370992 TI - Cranio-facial dysmorphism: experimental study in the mouse, clinical applications. AB - To obtain a better understanding of mandibulo-facial dysostosis and hemicraniofacial microsomia in man, the authors carried out a histologic and scanning electron microscope study of the facial malformations produced in mouse embryos by retinoic acid and methyl-triazene. The administration of 400 mg/kg 13 cis-retinoic acid (RA) to pregnant C57BL mice on day 9 of gestation produced anomalies of the cephalic extremity in the embryos resembling human mandibulo facial dysostosis. The 64 embryos collected presented hypoplasia of the branchial arches or the snout in 79% of cases, auricular anomalies in 47% and ophthalmic anomalies in 12.5%. Fourteen NMRI mice on day 10.5 of gestation were treated with 1.5 mg (0.5 mg/kg) methyl-triazene (Methyl). The 126 embryos collected had developed a very high percentage of micromandibles and anomalies of both embryonic ears (94.6% to 100%). Finally, although the facial anomalies produced by retinoic acid resemble the human mandibulo-facial dysostosis syndrome, no correlation was found between hemicraniofacial microsomia and the administration of methyl-triazene. PMID- 10370993 TI - Anatomic and radioanatomic study of the lateral genicular arteries: application to prevention of postoperative hemarthrosis after arthroscopic lateral retinacular release. AB - Arthroscopic lateral retinacular release can be complicated by hemarthrosis in 10 to 18% of cases. The vascular structures involved are the lateral vascular pedicles of the knee. This study examines the topography of these pedicles. Anatomic and radioanatomic studies carried out in 50 specimens defined the route of the vascular pedicles at the lateral aspect of the knee. From the measurements carried out, we noted the relative homogeneity of the routes taken by the different proximo-lateral vascular pedicles, which are highly vulnerable, and the variability of the disto-lateral arterial routes. A tracing-paper study identified two distinct routes for the disto-lateral vascular pedicle and evaluated the risk of injury to it in surgical approaches to the lateral aspect of the knee. Finally, the topographic data of the study suggest the possibility of preventive hemostasis of the proximo-lateral pedicle via a minimal approach close to the patella. Furthermore, it seems possible to avoid cutting the disto lateral pedicle if it is localised by cutaneous trans-illumination at the beginning of the operation. PMID- 10370994 TI - A new method of three-dimensional computer assisted reconstruction of the developing biliary tract. AB - A three-dimensional (3-D) computer assisted reconstruction of the biliary tract was performed in human and rat embryos at Carnegie stage 23 to describe and compare the biliary structures and to point out the anatomic relations between the structures of the hepatic pedicle. Light micrograph images from consecutive serial sagittal sections (diameter 7 mm) of one human and 16 rat embryos were directly digitalized with a CCD camera. The serial views were aligned automatically by software. The data were analysed following segmentation and thresholding, allowing automatic reconstruction. The main bile ducts ascended in the mesoderm of the hepatoduodenal ligament. The extrahepatic bile ducts: common bile duct (CD), cystic duct and gallbladder in the human, formed a compound system which could not be shown so clearly in histologic sections. The hepato pancreatic ampulla was studied as visualised through the duodenum. The course of the CD was like a chicane. The gallbladder diameter and length were similar to those of the CD. Computer-assisted reconstruction permitted easy acquisition of the data by direct examination of the sections through the microscope. This method showed the relationships between the different structures of the hepatic pedicle and allowed estimation of the volume of the bile duct. These findings were not obvious in two-dimensional (2-D) views from histologic sections. Each embryonic stage could be rebuilt in 3-D, which could introduce the time as a fourth dimension, fundamental for the study of organogenesis. PMID- 10370996 TI - Prospecting and evaluation of the anatomy sites on the Internet. AB - The Internet undoubtedly has an important part to play in medical teaching, generally speaking, and particularly in anatomy. We therefore undertook to survey, list, explore and study the various sites written in French or English devoted to anatomy on the Internet in order to evaluate them. Sites were identified from their URL address, by search engine or by hypertext links found in already listed sites. Useless and non-relevant sites were excluded. Fifty-two sites were selected and evaluated as to their navigability, illustrations, text and general presentation. Addresses of theses sites are available on a web page (http:/(/)www.rockefeller.univ-lyon1.fr/Anato mie-Lyon-Nord; section liens, section Galerie Virtuelle). This directory could be a very useful working tool for the use of teachers and students. It will be regularly updated. PMID- 10370995 TI - Subacromial space width changes during abduction and rotation--a 3-D MR imaging study. AB - The objectives of this study were to determine systematic changes of the normal subacromial space width during abduction and rotation, and to analyze the spatial relationship of the supraspinatus muscle with the acromion and clavicle. 12 healthy volunteers were imaged by an open MR scanner in 5 different positions of abduction and in 3 positions of rotation. After three dimensional (3D) reconstruction and 3D Euclidian distance transformation, the minimal spatial distances between the humerus and the acromion and the humerus and clavicle were computed. The minimal acromio-humeral distance decreased significantly from 30 degrees of abduction (mean 7.0 mm +/- 1.6 mm) to 120 degrees (mean 3.9 mm; +/- 1.8 mm; p < 0.0001). At 30 degrees, the minimal distance penetrated the supraspinatus, whereas at 120 degrees it was always located lateral to the supraspinatus tendon. At 90 degrees with internal rotation (7.6 mm, +/- 2.3 mm) the minimal acromio-humeral distance was larger than in neutral rotation (5.4 mm, +/- 2.3 mm) or external rotation (4.4 mm, +/- 2.2 mm; p < 0.05), but it penetrated the supraspinatus tendon at its most vulnerable part, reaching the acromion at its anterior inferior border. We conclude that the subacromial space width changes during abduction and rotation and that the supraspinatus is in closest contact to the anterior inferior border of the acromion in 90 degrees of abduction with 45 degrees internal rotation. These values obtained in volunteers can be used as a basis for further investigations in patients with the impingement syndrome. PMID- 10370997 TI - Variations and anomalies of the human orbital muscles. AB - The purpose of the study is to describe some rare and hitherto unreported uni- and bilateral anomalies of the orbital rectus muscles (duplication, triplication, accessory bellies, interrectal muscular bridges, false insertion) which were found by chance during the dissections of three cadavers (one adult, two fetuses). The levator palpebrae superioris muscles in the same specimens exhibited some variations (medial and lateral muscular slips) and anomalies (bipartite and unipartite levator). Cross-sections of interrectal bridges in the orbit of an adult were investigated histologically. Numerous nerves were distributed in the interstitium; groups of lipofuscin granules were found in all the myocytes. An attempt is made to explain these anomalies and variations through consideration of developmental and comparative anatomy. The relevance of these anomalies in coronal sections such as those acquired from CT and MRI is discussed. PMID- 10370999 TI - Multiple variations of the azygos venous system. AB - In a dissection performed in our department, we observed multiple variations of the azygos venous system. The hemiazygos vein was absent. The posterior 8th, 9th, and 10th intercostal veins united and their common trunk crossed the vertebral column obliquely lying anterior to the aorta and posterior to the esophagus and opening into the azygos vein at the level of T7-T8 vertebrae. The 7th left posterior intercostal vein also crossed the column anteriorly and joined the common trunk. The present report identifies the variable positions and courses of the veins related to the azygos system. It is important to keep in mind that different courses of the azygos system do exist, so that extra caution is required during surgery of the mediastinum and also in appropriately interpreting the radiographs. PMID- 10370998 TI - Left renal vein variations. AB - The highly complex embryological development of the left renal vein compared to its right counterpart results in greater variations which are clinically significant. The study aimed to identify these variations and to document its incidence. Cadaveric study: 153 kidney pairs were harvested en bloc, dissected, 100 resin casts prepared and 53 plastinated; renal venography performed on further 58 adults and 20 foetal cadavers. Clinical study: (retrospective analysis): a) radiological study, 104 renal venograms; b) live related renal transplantation, 148 donor left kidneys; c) abdominal aortic aneurysm surgery, 525 patients. Total sample size: 1008. Renal collars observed in 0.3%; retro aortic vein 0.5%; additional veins 0.4%; posterior primary tributary 23.2%, (16.7% Type IB; 6.5% Type IIB, cadaveric series, only). Our results differ significantly in incidence to that reported in the literature: renal collar 0.2 30%; retro-aortic vein 0.8-7.1%; additional renal vein 0.8-6%. Variations are clinically silent and remain unnoticed until discovered during venography, operation or autopsy. To a transplant surgeon, morphology acquires special significance, since variations influence technical feasibility of operation. Prior knowledge of circum-aortic vein is important when blood samples from suprarenal or renal veins are collected. Collar may provide developed collateral pathway immediately after surgery if renal interruption planned without awareness of its presence. Variations restrict availability of vein for mobilisation procedures. In aortic aneurysm repair, retro-aortic vein is important. During retroperitoneal surgery, the surgeon may visualise a pre-aortic vein but be unaware of an additional retroaortic component or a posterior primary tributary, and may avulse it while mobilising the kidney or clamping the aorta. PMID- 10371000 TI - Resin-modified glass ionomer cement restorations in primary molars. AB - The aim of this multicenter study was to evaluate the 3-year performance of a resin-modified glass ionomer cement in primary molars. A total of 174 class II restorations were placed in 85 children by 6 dentists. The restorations were evaluated during a 3-year period using slightly modified USPHS criteria (van Dijken 1986). Of the 174 restorations 161 were evaluated after 1 year, 121 after 2 years and 68 after 3 years. A total of 81, restorations exfoliated or was extracted during the study. The cumulative failure rate after 1, 2 and 3 year was 8.1%, 11.7% and 19.8% respectively. The main reasons for failure were secondary caries and loss of retention. PMID- 10371001 TI - Activation of human B-lymphocytes by Prevotella intermedia. AB - Black pigmented Gram negative Prevotella intermedia is a potential pathogenic bacterium in periodontal disease. Lipopolysaccharides (LPS) from Gram negative bacteria are the main antigens that stimulate antibody production and cytokine synthesis. Many reports indicate a strong correlation of specific antibodies of the immunoglobulin G (IgG) class with antigens of periopathogenic bacteria in periodontal disease. The aim of this study was to measure lymphocyte proliferation and immunoglobulin (IgA, IgG and IgM) production from B-lymphocytes in response to P. intermedia stimulation. P. intermedia was originally isolated from human periodontal pockets by means of a gingival crevice lavage method. The periodontal pocket was washed with salt solution and the solution was then aspirated into a cannula for further culturing in the laboratory. The identification of P. intermedia was made on Brucella agar. Lymphocytes were isolated from human peripheral blood. P. intermedia was added to the lymphocytes in concentrations from 0.005% to 0.1%. The immunoglobulin production was measured by enzyme-linked immunosorbent assay (ELISA). The results showed that IgA and IgG were produced in response to stimulation with P. intermedia. The IgG response was dose-dependent. No stimulation of IgM production was obtained. It is concluded from the present observations that P. intermedia can stimulate proliferation of lymphocytes and production of IgA and IgG. PMID- 10371002 TI - Oral health in pre-school children living in Sweden. Part III--A longitudinal study. Risk analyses based on caries prevalence at 3 years of age and immigrant status. AB - A decreasing proportion of children require operative treatment each year and an increasing proportion of children will at recall require no operative care. This would provide resources to give individualised prevention to children at risk for developing new carious lesions. From 3 years of age, however, the prevalence of caries increases up to the age when the primary dentition exfoliates, even though the children attend regular dental service programmes yearly, and for some children dental caries remains a significant problem. The purpose of the present study was to describe the dental health of a group of 6-year-old children living in Sweden, with special reference to caries prevalence at 3 years of age and to immigrant status. At 6 years of age, 45% of the children were free of initial and manifest carious lesions in the primary cuspids and molars. The mean caries increment between 3 and 6 years was 0.9 tooth surfaces for children who were caries free at 3 years of age compared to 4.5 tooth surfaces for children with manifest carious lesions at the same age. The mean caries increment from 3 to 6 years was 1.3 in the non-immigrant group and 3.6 in the immigrant group. For the majority of the children in this study, current preventive dental care seems sufficient. However, for about one-third of the children (children with carious lesions at 3 years of age and/or immigrant status), current caries preventive programme used in the Public Dental Service are inadequate. PMID- 10371003 TI - Therapeutic jaw exercises and interocclusal appliance therapy. A comparison between two common treatments of temporomandibular disorders. AB - From a total of 1344 consecutive patients referred to a TMD clinic, twenty-six patients fulfilled the strict inclusion criterias of TMD of mainly muscular origin. Half of the patients were assigned to receive treatment with an interocclusal appliance, the treatment being performed by a dentist. The other half was instructed to perform individualized therapeutic jaw exercises, and this treatment was managed by a dental assistant. The treatment result was evaluated after six months. The two treatments had a positive and equal effect upon both signs and symptoms of TMD. A further follow-up by questionnaire one to four years after the final clinical examination showed a lasting treatment result in most patients. Many patients, however, continued to perform jaw exercises and/or to wear their appliances. This indicates that these two treatments are mostly symptomatic and not causal. The conclusion of the present investigation is that therapeutic jaw exercises, managed by a dentist or a dental assistant, is a cost effective treatment with a prognosis comparable to a treatment with an interocclusal appliance and can thus be recommended as the first therapy of choice in patients with TMD of mainly muscular origin. PMID- 10371004 TI - The influence of stabilisation appliance therapy and other factors on the treatment outcome in patients with temporomandibular disorders of arthrogeneous origin. AB - The aim of this study was to investigate if the difference in treatment outcome between patients provided with a stabilisation appliance and a control appliance was due to the treatment and/or other factors in patients with temporomandibular disorders (TMD) of arthrogeneous origin. Sixty patients were assigned to two equally sized groups: a treatment group, treated with a stabilisation appliance, and a control group given a control appliance. Thirteen possible background variables for the treatment outcome were correlated to changes in severity of temporomandibular joint (TMJ) pain on a verbal scale in the two patient groups. The logistic regression analyses revealed that, after correcting for the background variables, stabilisation appliance treatment was a strong explanatory factor for a positive treatment outcome, with a significance of P = 0.0013 compared to patients belonging to the control group. Background variables of significant importance for the treatment outcome were male sex (positive) (P = 0.0268), and severe or very severe TMJ pain (negative) (P = 0.0034). These findings indicate that not only the treatment with a stabilisation appliance but also sex and the intensity of the TMJ pain before treatment might influence the treatment outcome in patients with TMD of arthrogeneous origin. PMID- 10371005 TI - Rinderpest vaccination and the incidence and development of trypanosomosis in cattle. AB - An investigation was made into whether recent vaccination of cattle with tissue culture rinderpest virus would cause immunosuppression and lead to more frequent or more severe infection with trypanosomes in animals grazing in tsetse-infested areas. Herds of cattle on Galana Ranch in Kenya were divided, with approximately half of each herd being vaccinated with tissue culture rinderpest virus strain Kabete 'O', while the rest remained unvaccinated. The herds were then exposed to the risk of natural infection with trypanosomes on the ranch. Three experiments were performed during different seasons. Infections with Trypanosoma congolense and Trypanosoma vivax were frequently detected but there was no evidence that vaccinated animals were more likely to acquire trypanosome infections or to show a more severe disease than unvaccinated cattle. It is concluded that tissue culture rinderpest vaccine does not cause immunosuppression and can safely be used in cattle likely to be exposed to tsetse flies and trypanosomosis. PMID- 10371007 TI - Prevalence and distribution of animal trypanosomosis on Buvuma islands in Lake Victoria, Uganda. PMID- 10371006 TI - Susceptibility of local Nigerian and exotic chickens to infectious bursal disease by contact exposure. AB - One hundred 6-week-old susceptible cockerels were inoculated with a pathogenic strain of infectious bursal disease virus (IBDV) and kept in the same pen as 100 each of 6-week-old pullets, local chickens and broilers. The cockerels developed depression and diarrhoea on day 3 post inoculation (PI) and most of the pullets and some of the local chickens and broilers showed similar signs on day 4 PI. Loss in weight was severe and similar in the pullets and local chickens, being significantly greater than that in the broilers from days 3-11 PI. The total mortality was 85%, 66.7%, 30% and 20% for the pullets, cockerels, local chickens and broilers, respectively. The lesions were more severe in the pullets and local chickens than in the broilers. IBDV antigen and antibody were detected, respectively, in all the bursal and serum samples from the infected chickens tested. The contact exposure method used in this study simulates better what happens in nature than inoculation with IBDV. The reduced mortality observed among the local chickens, compared with that (61.5%) seen in earlier studies using intraocular inoculation of IBDV, may have been due to behavioural differences that tend to result in their ingesting a relatively low dose of the virus. PMID- 10371008 TI - Improvement of cattle productivity through rapid alleviation of African animal trypanosomosis by integrated disease management practices in the agropastoral zone of Yale, Burkina Faso. AB - Investigations to identify the causes of high mortalities in cattle in the agropastoral zone (ZAP) of Yale started in March 1993. African animal trypanosomosis (AAT) was found to be the major constraint, with incidence rates exceeding 30%, justifying a tsetse control programme, which started in March/ April 1994. The treatment of all cattle at bimonthly intervals with deltamethrin 1% pour on and the display of 1500 insecticide impregnated targets during the 6 months of the dry season each year helped to reduce the tsetse populations (Glossina tachinoides and G. morsitans submorsitans) by more than 90%. In less than 7 months, the incidence of AAT dropped below 5% and remained there throughout the intervention until June 1996, in spite of an increase to 3 months in the interval between the treatments. Mean PCV values increased significantly from 26.5-30.9%, before, to 30.7-36.3% during the intervention. The improvement in the overall health resulted in a resumption in fertility and milk production, allowing the sale of dairy products in Leo, thus creating a gross income of about $US3/day for the Fulani women. PMID- 10371009 TI - Perceived constraints to privatization of delivery of veterinary services in Ghana. AB - Ghana is on the verge of privatizing selected activities in the delivery of animal health services. However, various constraints are being encountered. The aim of this paper is to identify these constraints so as to help find and solutions to them. Questionnaires were administered to veterinarians in Ghana to elicit their responses on various issues concerning privatization. A significant proportion (61%) of government veterinarians, who formed 94% of the respondents, were unwilling to go into private practice. Among the reasons given were that private practice was too risky, that farmers were unwilling or unable to pay for services, that capital to start practices was lacking and that the societal value for animals was low. Also, low livestock densities in many areas and the absence of commercial livestock farming were perceived as deterrents to the sustainability of private practice. Furthermore, the poor macroeconomic environment of high inflation, high interest rates and unstable currency discouraged investment. If privatization of veterinary services is to succeed in Ghana, these perceptions have to be addressed and solutions found, since veterinarians are the targets of the privatization process. PMID- 10371010 TI - Calliandra calothyrsus as a supplement for milk production in the Kenya highlands. AB - Two on-farm experiments and one on-station observation were conducted between July 1994 and September 1995 to study the effect of supplementation with fresh fodder of Calliandra calothyrsus on milk production from grade Friesian and Ayrshire cows in the second trimester of their lactations. The cattle were kept under zero-grazing systems on small farms in the coffee-based land use system at altitudes of 1500 to 1800 m on the slopes of Mt Kenya. These cows form a pivotal part of the farming system since they produce both milk for sale and manure for crop production. Milk production is normally in the region of 10 kg/cow per day when the animals are fed on a diet based on Napier grass and crop residues, together with 2-4 kg day of commercial concentrate. In terms of milk production, 3 kg of fresh calliandra had the same effect on yield as 1 kg of additional dairy meal and, at normal production levels, the effects of the two supplements were strictly additive. Calliandra had a marked positive effect (about a 10% increase) on the butterfat content of the milk, a factor that was highly valued by farmers, even though institutional buyers as yet offer no premium price for milk quality. The average small farm can produce enough calliandra fodder to supplement two dairy cows and some additional small stock from relatively underutilized niches along the farm perimeter and terrace risers, without any adverse effect on current levels of crop production. PMID- 10371011 TI - Herd-specific and age-specific seroprevalence of Neospora caninum in 14 British dairy herds. AB - All the cattle in 14 dairy herds in England were tested for Neospora caninum specific antibodies with a commercial ELISA. Three of the herds had had sporadic abortions, eight had had endemic abortions and three had had epidemic abortions associated with N caninum before the study. Of 4295 cattle tested, 17.1 per cent were seropositive and the herd-specific prevalence ranged from 7.3 per cent to 44.8 per cent. No significant effect of either herd size (P = 0.988), endemic (P = 0.869) or epidemic (P = 0.138) patterns of abortion on herd-specific prevalence was found by using logistic regression analysis. There was no evidence in any herd of a significant increase in prevalence with age; the prevalence in seven-to 12-month-old cattle was not significantly different (P > 0.400) from the prevalence in older cattle, except that there was a significantly lower prevalence (P = 0.017) in 13-to 24-month-old cattle. The results of this study are consistent with vertical transmission being the major route of N caninum transmission in these herds. PMID- 10371012 TI - Dynamic adrenal function testing in eight dogs with hyperadrenocorticism associated with adrenocortical neoplasia. AB - The results of adrenocorticotropin (ACTH) stimulation and low-dose dexamethasone suppression tests (LDDST) were evaluated retrospectively in eight dogs with clinical signs of hyperadrenocorticism arising from functional adrenocortical tumours, and compared with the results from 12 dogs with confirmed pituitary dependent hyperadrenocorticism (PDH). The post-ACTH cortisol concentration in the dogs with adrenocortical tumours ranged from 61 to 345-6 nmol/litre (median 251.5 nmol/litre) and they were within the reference range (150 to 450 nmol/litre) in five and unexpectedly low (< 150 nmol/litre) in three dogs. Both the basal and post-ACTH cortisol concentrations were significantly lower in the dogs with adrenocortical neoplasia than in the dogs with PDH. Eight hours after the LDDST, only two of six dogs with adrenocortical tumours had a cortisol concentration above 30 nmol/litre, and the median resting, three, and eight-hour cortisol concentrations were 31.5, 23.0, and 22.7 nmol/litre respectively. There was no significant cortisol suppression during the LDDST, although interpretation was complicated by the low cortisol concentrations, but two dogs showed a pattern of apparent suppression. Two dogs with adrenal tumours showed a diagnostically significant increase in 17-OH-progesterone concentration in response to ACTH although their cortisol concentrations did not increase greatly. These results differ from previous reports of the response of functional adrenal tumours to dynamic endocrine tests. PMID- 10371013 TI - Experimental Brucella ovis infection in pregnant ewes. AB - Forty yearling Brucella-free ewes were inoculated with Brucella ovis by the conjunctival route in mid or late first pregnancy. Only a few ewes excreted B ovis during pregnancy and gave birth to stillborn lambs, but most of them excreted the organism at lambing or during lactation. One of the 11 lambs which were born alive but died before they were weaned was found to be infected postmortem. In contrast, none of the 46 surviving lambs which were reared in isolation until adulthood, was found to be infected. At weaning, the 40 ewes were mated again with five Brucella-free rams. Although many of the ewes excreted B ovis, none of the rams was found to be infected when necropsied after mating. Most of the ewes that became pregnant, all having excreted B ovis during their first pregnancy, cleared the infection during the second pregnancy. However, three remained persistently infected and excreted B ovis in their milk throughout the second lactation. None of the lambs born to these three ewes was found to be infected when necropsied at weaning. PMID- 10371014 TI - Dental wear in horses in relation to the microhardness of enamel and dentine. AB - The microhardness of enamel, primary dentine and secondary dentine was determined in the incisor teeth of 39 horses of three different breeds, trotter horses, Belgian draft horses and Arab horses. Using a microhardness tester fitted with a Knoop diamond indenter, the overall Knoop Hardness Number was determined for each tissue, and the influence of breed and age on the hardness were evaluated. Enamel and secondary dentine were significantly harder in Arab horses than in trotters and Belgian draft horses, but there were no significant differences between draft horses and trotter horses in the hardness of their enamel and dentine. PMID- 10371015 TI - Metaphyseal osteomyelitis in a Labrador retriever. PMID- 10371016 TI - Persistence of the efficacy of a novel injectable ivermectin formulation and injectable doramectin against gastrointestinal nematodes in cattle. PMID- 10371017 TI - Displaced abomasum in dairy cattle. PMID- 10371018 TI - Probing the structure of multi-stranded guanine-rich DNA complexes by Raman spectroscopy and enzymatic degradation. AB - The multi-stranded DNA complexes formed by the oligonucleotides d(T15G4T2G4), Tel, and d(T15G15), TG, were examined by nuclease digestion and Raman spectroscopy. Both Tel and TG can aggregate to form structures consisting of multiple, parallel-oriented DNA strands with two independent structural domains. Overall, the structures of the TG and Tel aggregates appear similar. According to the Raman data, the majority of bases are in C2'-endo/anti conformation. The interaction of guanines at the 3'-ends in both complexes stabilizes the complexes and protects them from degradation by exonuclease III. The 5'-extensions remain single-stranded and the thymines are accessible to single-strand-specific nuclease digestion. The extent of enzymatic cleavage at the junction at the 5' end of the 15 thymines implies a conformational change between this part of the molecule and the guanine-rich region. The differential enzymatic sensitivity of the complexes suggests there are variations in backbone conformations between TG and Tel aggregates. TG aggregates were more resistant to digestion by DNase I, Mung Bean nuclease, and S1 nuclease than Tel complexes. It is proposed that the lower DNase I sensitivity may be partly due to the more stable backbone exhibited by TG than Tel complexes. Structural uniformity along the guanine core of TG is suggested, as there is no indication of structural discontinuities or protected sites in the guanine-rich regions of TG aggregates. The lower extent of digestion by Mung Bean nuclease at the 3' end implies that these bases are inaccessible to the enzyme. This suggests that there is minimal fraying at the ends, which is consistent with the extreme thermal stability of the TG aggregates. PMID- 10371020 TI - Reaction-diffusion microtubule concentration patterns occur during biological morphogenesis. AB - Reaction-diffusion processes can lead to a macroscopic concentration pattern from an initially homogeneous solution, and thus provide a physical-chemical mechanism for biological pattern formation and morphogenesis. The central prediction of reaction-diffusion theory is that the patterns contain periodic concentration variations in some of the reactives. Microtubules assembled in vitro spontaneously self-organise and form stationary striped macroscopic structures. In agreement with reaction-diffusion theory. Here we show, in agreement with reaction-diffusion theory, that these preparations contain substantial microtubule concentration variations. Similar striped microtubule patterns arise during Drosophila embryogenesis. A characteristic of these patterns is their dependence on sample dimensions. In Drosophila eggs shortened by ligation, we found that the microtubule pattern varied with egg fragment length in the same way as the in vitro microtubule pattern varied with sample length, and as expected from theory. This is evidence that reaction-diffusion structures occur during Drosophila morphogenesis. PMID- 10371019 TI - Three-photon excitation of N-acetyl-L-tyrosinamide. AB - We observed emission from the tyrosine derivative N-acetyl-L-tyrosinamide (NATyrA) when excited with the fundamental output of a femtosecond Ti:Sapphire laser from 780 to 855 nm. The dependence on incident laser power indicates a three-photon process. The emission spectra and intensity decay in glycerol-water (30:70) at 5 degrees C were found to be identical for one- and three-photon excitation. Also the excitation spectrum of three-photon-induced fluorescence of NATyrA corresponds to the one-photon excitation spectrum. The time-zero or fundamental anisotropy spectrum was reconstructed from the frequency-domain anisotropy decays. The three-photon anisotropies are similar or larger than the one-photon anisotropies. These three-photon anisotropies are surprising given the near zero values known for tyrosine with two-photon excitation. The observations indicate that one- and three-photon excitation directly populates the same singlet excited states(s). However, the origin of the anisotropies with multi photon excitation of tyrosine remain unclear and unpredictable. PMID- 10371021 TI - Simulation of the distribution and diffusion of a rigid amphipathic particle embedded in a model membrane. AB - We simulate, by Brownian dynamics, the distribution, orientation and diffusion of a rigid molecule, represented as a dumbbell, with amphipathic nature, embedded in a model membrane. The significant features of a biological membrane are reproduced by means of a Maier-Saupe orienting potential, an enclosing potential and a lipophobic potential. We also evaluate the equilibrium quantities, such as order parameter, and dynamic features, such as rotational or translational diffusivity, of the embedded molecule in terms of the system parameters and compare the obtained results with those obtained from model independent theory. PMID- 10371022 TI - Electrochemical studies of cytochrome c disulfide at gold electrodes. AB - The electrochemistry of disulfide in cytochrome c on gold electrodes was reported. The observed electrochemical response was used to explain why the electrochemical reaction of cytochrome c is irreversible at gold electrodes. Disulfide bonds in cytochrome c were strongly adsorbed onto the surface of gold electrodes and caused slow rate of electron transfer of the heme group. It was found that the presence of disulfides in cytochrome c was responsible for the lack of electrochemical response of the heme group on a gold electrode. The mechanisms for this effect were studied using electrochemistry and photoelectron spectroscopy. PMID- 10371023 TI - Correlation of activity of 2-(X-benzyloxy)-4,6-dimethoxyacetophenones with topological indices and with the Hansch equation. AB - The antispasmodic activities of the 2-(X-benzyloxy)-4,6-dimethoxyacetophenones (X = H, 4'-F, 4'-NO2, 4'-CH3, 4'-Cl, 3',4'-(CH3)2, 4'-OCH3, 4'-Br, 4'-C(CH3)3, 4' OCH2C6H5) against acetylcholine-induced contraction of the guinea pig ileum were correlated with different topological indices. Good correlations were obtained through a simple regression equation with electrotopological state indices (Si) for the carbon atoms S(C1) and S(C6). Using multiple linear regression with two variables the best correlations were obtained with carbons in the 6- and 1 positions with sigma. Such results indicate that the corresponding carbon atoms play an important role in the biological activity. The equation of Hansch showed that the activity of these compounds increases when the ring substituent in the benzyloxy group are more highly electron-releasing and hydrophobic. PMID- 10371024 TI - Serum beta-endorphin level in patients with depression on fluvoxamine. AB - The main interest of the present study was to determine possible alternations in beta-endorphin serum levels in healthy volunteers and in patients with depression, as well as changes in beta-endorphin serum levels caused by fluvoxamine treatment. Fluvoxamine maleate (Fevarin) was administered orally at a dose of 200 mg/day for 4 weeks. The serum levels of beta-endorphin were lower in patients with 'nonendogenous' depression (104.68 +/- 5.29 pg/ml) and those with 'endogenous' depression (36.34 +/- 2.23 pg/ml) than in healthy volunteers (125.19 +/- 1.64 pg/ml). The endogenously depressed patients had significantly lower beta endorphin levels than the nonendogenous patients. A 4-week treatment of fluvoxamine (200 mg/day) caused a statistically significant increase in beta endorphin serum levels in all patients (nonendogenous depression 132.10 +/- 2.38 pg/ml and endogenous depression 50.09 +/- 2.45 pg/ml) in comparison to values found before the onset of the therapy. The efficacy of fluvoxamine was 11.0 (+/- 9.0) evaluated by the Hamilton Rating Scale for Depression (HAMD) in the patients with a diagnosis of depression. These results indicate that determination of beta endorphin serum levels could be a valuable laboratory test in the diagnosis of depression. PMID- 10371026 TI - Electrochemical behaviour of Venlafaxine and its determination in pharmaceutical products using square wave voltammetry. AB - An electrochemical method for the determination of Venlafaxine in pharmaceutical formulations recently available on the European market is described. The electrochemical oxidation of Venlafaxine was studied at a HMDE electrode over a wide pH range (1.9-10.0) of buffered aqueous solutions using different potential sweep techniques. The results obtained showed that the best definition of the analytical signals was found in boric acid/potassium tetrahydroxoborate buffer at pH 8.7 using anodic stripping square wave voltammetry. Recovery trials were performed to assess the accuracy of results and these were compared to those provided by a HPLC technique according to the method described in literature and to the features of the pharmaceutical formulations. A relative deviation of < 0.2% was obtained. With the developed methodology, the limit of detection was 0.124 mg/l. PMID- 10371025 TI - Photochemical and photobiological studies on methylthioangelicins. AB - 4,4',5'-Trimethyl-1'-thioangelicin (1) and 4,6,4',5'-tetramethyl-1'-thioangelicin (2), two newly synthesised isosters of furocoumarins having a sulfur atom in their five-membered ring, were studied in terms of interactions with DNA, both in the ground state and after UVA light absorption. The compounds were able to intercalate the macromolecule and to photobind efficiently, forming C4 cycloadducts with thymine. The antiproliferative effect of this binding was shown in Ehrlich and HeLa cells and by T2 phage inactivation. Tests on Salmonella typhimurium indicated low mutagenic activity. In particular, compound 1 has photobiological activity comparable with that of 4,6,4'-trimethylangelicin, but is less mutagenic. PMID- 10371027 TI - Synthesis of some new biologically active thiadiazolotriazinones--Part II. AB - 4-Amino-6-phenyl/methyl-3-mercapto-1,2,4-triazin-5(4H)-ones (1) are condensed with an aromatic carboxylic acid, aryloxyacetic acid or anilinoacetic acid (2), to yield 7-substituted-3-phenyl/methyl-4H-1,3,4-thiadiazolo-[2,3-c]-1,2,4-+ ++triazin-4- ones (3). Phosphorus oxychloride is used as a cyclizing agent. All the synthesized compounds are screened for their antibacterial activities against S. aureus, E. coli, P. aeruginosa and G. bacillus. PMID- 10371028 TI - Pyrrolo[3,2-c]quinoline derivatives: a new class of kynurenine-3-hydroxylase inhibitors. AB - A series of pyrrolo[3,2-c]quinoline derivatives were synthesised and evaluated as inhibitors of selected enzymes of the kynurenine pathway. 7-Chloro-3-methyl-1H pyrrolo[3,2-c]quinoline-4-carboxylic acid (7a) was found to be a relatively potent and selective inhibitor of kynurenine-3-hydroxylase (KYN-3-OHase). A molecular modelling study showed a good superimposition of 7a with PNU-156561 and kynurenine the natural substrate of KYN-3-OHase. PMID- 10371029 TI - Synthesis of 3,6,7-substituted-quinoxalin-2-ones for evaluation of antimicrobial and anticancer activity. Part 2. AB - A new set of 35 3-alkyl and 3-ethoxycarbonylalkyl 6- and/or 7-substituted-2 quinoxalinones was prepared and submitted to a preliminary in vitro investigation of their antimicrobial, anticancer and anti-HIV activities. Results are referred. PMID- 10371030 TI - Preparation and biological evaluation of 6/7-trifluoromethyl(nitro)-, 6,7 difluoro-3-alkyl (aryl)-substituted-quinoxalin-2-ones. Part 3. AB - A new series of quinoxalinones 6/7-trifluoromethyl or nitro- and 6,7-difluoro substituted bearing various side-chains (alkyl, halogenoalkyl, benzyl and phenyl groups) at C-3 of the ring system was synthesized and submitted to preliminary in vitro evaluation for antibacterial, antifungal, antimycobacterial, anticancer and anti-HIV activities. Results of these screenings showed that compounds 23-28 exhibited a good inhibition activity against various strains of Candida. Compound 24 showed also an interesting in vitro anticancer activity. PMID- 10371031 TI - Synthesis and anticonvulsant activity of new 2,3-benzodiazepines as AMPA receptor antagonists. AB - Novel 1-aryl-3,5-dihydro-7,8-methylenedioxy-4H-2,3-benzodiazepine-4-ones (12a-j) were prepared and their anticonvulsant effects were evaluated by using various models of experimental epilepsy. The seizures were evoked both by means of auditory stimulation in DBA/2 mice and by pentylenetetrazole or maximal electroshock in Swiss mice. Some of these compounds possess marked anticonvulsant properties in all tests employed. Compounds 12 antagonise seizures induced by AMPA in analogy to the structurally-related 1-(4'-aminophenyl)-4-methyl-7,8 methylenedioxy-5H-2,3- benzodiazepine (1) (GYKI 52466), a well-known non competitive AMPA-receptor antagonist. On the other hand, these novel 2,3 benzodiazepines exhibit anticonvulsant properties that are not affected by flumazenil, but are reversed by aniracetam. In addition, when compared to model compound 1, compounds 12 show a longer-lasting anticonvulsant activity and a lower toxicity. A structure-activity relationship study carried out on compounds 12 as well as analogous 7,8-dimethoxy derivatives 2 offers an approach for designing more potent agents. PMID- 10371032 TI - Synthesis and antibacterial activity of 2-aryl-2,5-dihydro-3(3H)-oxo pyridazino[4,3-b]indole-4-carboxylic acids. AB - The in vitro antibacterial and antifungal activities of a series of pyridazinoindolonic acids II against some selected representative of Gram positive, Gram-negative bacteria and fungi have been investigated. Some interesting observations among the structural features necessary for high antibacterial activity are presented and discussed. PMID- 10371033 TI - Ribozymes: the characteristics and properties of catalytic RNAs. AB - Ribozymes, or catalytic RNAs, were discovered a little more than 15 years ago. They are found in the organelles of plants and lower eukaryotes, in amphibians, in prokaryotes, in bacteriophages, and in viroids and satellite viruses that infect plants. An example is also known of a ribozyme in hepatitis delta virus, a serious human pathogen. Additional ribozymes are bound to be found in the future, and it is tempting to regard the RNA component(s) of various ribonucleoprotein complexes as the catalytic engine, while the proteins serve as mere scaffolding- an unheard-of notion 15 years ago! In nature, ribozymes are involved in the processing of RNA precursors. However, all the characterized ribozymes have been converted, with some clever engineering, into RNA enzymes that can cleave or modify targeted RNAs (or even DNAs) without becoming altered themselves. While their success in vitro is unquestioned, ribozymes are increasingly used in vivo as valuable tools for studying and regulating gene expression. This review is intended as a brief introduction to the characteristics of the different identified ribozymes and their properties. PMID- 10371034 TI - 3'-End processing of pre-mRNA in eukaryotes. AB - 3'-Ends of almost all eukaryotic mRNAs are generated by endonucleolytic cleavage and addition of a poly(A) tail. In mammalian cells, the reaction depends on the sequence AAUAAA upstream of the cleavage site, a degenerate GU-rich sequence element downstream of the cleavage site and stimulatory sequences upstream of AAUAAA. Six factors have been identified that carry out the two reactions. With a single exception, they have been purified to homogeneity and cDNAs for 11 subunits have been cloned. Some of the cooperative RNA-protein and protein protein interactions within the processing complex have been analyzed, but many details, including the identity of the endonuclease, remain unknown. Several examples of regulated polyadenylation are being analyzed at the molecular level. In the yeast Saccharomyces cerevisiae, sequences directing cleavage and polyadenylation are more degenerate than in metazoans, and a downstream element has not been identified. The list of processing factors may be complete now with approximately a dozen polypeptides, but their functions in the reaction are largely unknown. 3'-Processing is known to be coupled to transcription. This connection is thought to involve interactions of processing factors with the mRNA cap as well as with RNA polymerase II. PMID- 10371035 TI - RNA editing. AB - The term RNA editing describes those molecular processes in which the information content is altered in an RNA molecule. To date such changes have been observed in tRNA. rRNA and mRNA molecules of eukaryotes, but not prokaryotes. The demonstration of RNA editing in prokaryotes may only be a matter of time, considering the range of species in which the various RNA editing processes have been found. RNA editing occurs in the nucleus, as well as in mitochondria and plastids, which are thought to have evolved from prokaryotic-like endosymbionts. Most of the RNA editing processes, however, appear to be evolutionarily recent acquisitions that arose independently. The diversity of RNA editing mechanisms includes nucleoside modifications such as C to U and A to I deaminations, as well as non-templated nucleotide additions and insertions. RNA editing in mRNAs effectively alters the amino acid sequence of the encoded protein so that it differs from that predicted by the genomic DNA sequence. PMID- 10371036 TI - Dynamics of translation on the ribosome: molecular mechanics of translocation. AB - The translocation step of protein elongation entails a large-scale rearrangement of the tRNA-mRNA-ribosome complex. Recent years have seen major advances in unraveling the mechanism of the process on the molecular level. A number of intermediate states have been defined and, in part, characterized structurally. The article reviews the recent evidence that suggests a dynamic role of the ribosome and its ligands during translocation. The focus is on dynamic aspects of tRNA movement and on the role of elongation factor G and GTP hydrolysis in translocation catalysis. The significance of structural changes of the ribosome induced by elongation factor G as well the role of ribosomal RNA are addressed. A functional model of elongation factor G as a motor protein driven by GTP hydrolysis is discussed. PMID- 10371037 TI - Selenocysteine inserting tRNAs: an overview. AB - One of the recent discoveries in protein biosynthesis was the finding that selenocysteine, the 21st amino acid, is cotranslationally inserted into polypeptides under the direction of a UGA codon assisted by a specific structural signal in the mRNA. The key to selenocysteine biosynthesis and insertion is a special tRNA species, tRNA(Sec). The formation of selenocysteine from serine represents an interesting tRNA-mediated amino acid transformation. tRNA(Sec) (or the gene encoding it) has been found over all three domains of life. It displays a number of unique features that designate it a selenocysteine-inserting tRNA and differentiate it from canonical elongator tRNAs. Although there are still some uncertainties concerning the precise secondary and tertiary structures of eukaryal tRNA(Sec), the major identity determinant for selenocysteine biosynthesis and insertion appears to be the 13 bp long extended acceptor arm. In addition the core of the 3D structure of these tRNAs is different from that of class II tRNAs like tRNA(Sec). The biological implications of these structural differences still remain to be fully understood. PMID- 10371038 TI - mRNA degradation in bacteria. AB - Messenger RNAs in prokaryotes exhibit short half-lives when compared with eukaryotic mRNAs. Considerable progress has been made during recent years in our understanding of mRNA degradation in bacteria. Two major aspects determine the life span of a messenger in the bacterial cell. On the side of the substrate, the structural features of mRNA have a profound influence on the stability of the molecule. On the other hand, there is the degradative machinery. Progress in the biochemical characterization of proteins involved in mRNA degradation has made clear that RNA degradation is a highly organized cellular process in which several protein components, and not only nucleases, are involved. In Escherichia coli, these proteins are organized in a high molecular mass complex, the degradosome. The key enzyme for initial events in mRNA degradation and for the assembly of the degradosome is endoribonuclease E. We discuss the identified components of the degradosome and its mode of action. Since research in mRNA degradation suffers from dominance of E. coli-related observations we also look to other organisms to ask whether they could possibly follow the E. coli standard model. PMID- 10371039 TI - Function, mechanism and regulation of bacterial ribonucleases. AB - The maturation and degradation of RNA molecules are essential features of the mechanism of gene expression, and provide the two main points for post transcriptional regulation. Cells employ a functionally diverse array of nucleases to carry out RNA maturation and turnover. Viruses also employ cellular ribonucleases, or even use their own in their reproductive cycles. Studies on bacterial ribonucleases, and in particular those from Escherichia coli, are providing insight into ribonuclease structure, mechanism, and regulation. Ongoing biochemical and genetic analyses are revealing that many ribonucleases are phylogenetically conserved, and exhibit overlapping functional roles and perhaps common catalytic mechanisms. This article reviews the salient features of bacterial ribonucleases, with a focus on those of E. coli, and in particular, ribonuclease III. RNase III participates in a number of RNA maturation and RNA decay pathways, and is regulated by phosphorylation in the T7 phage-infected cell. Plasmid and phage RNAs, in addition to cellular transcripts, are RNase III targets. RNase III orthologues occur in eukaryotic cells, and play key functional roles. As such, RNase III provides an important model with which to understand mechanisms of RNA maturation, RNA decay, and gene regulation. PMID- 10371040 TI - Ribonuclease P: the diversity of a ubiquitous RNA processing enzyme. AB - Ribonuclease P is the endonuclease required for generating the mature tRNA 5' end. The ribonucleoprotein character of this enzyme has now been proven in most organisms and organelles. Exceptions, however, are still the chloroplasts, plant nuclei and animal mitochondria where no associated RNAs have been detected to date. In contrast to the known RNA subunits, which are fairly well-conserved in size and structure among diverse phylogenetic groups, the protein contribution to the holoenzyme is highly variable in size and number of the individual components. The structure of the bacterial protein component has recently been solved. In contrast, the spatial arrangement of the multiple subunits in eukaryotic enzymes is still enigmatic. Substrate requirements of the enzymes or their catalytic RNA subunits are equally diverse, ranging from simple single domain mimics to an almost intact three-dimensional structure of the pre-tRNA substrate. As an example for an intermediate in the enzyme evolution, ribonuclease P from the Cyanophora paradoxa cyanelle will be discussed in more detail. This enzyme is unique, as it combines cyanobacterial and eukaryotic features in its function, subunit composition and holoenzyme topology. PMID- 10371042 TI - Detection of low levels of flunitrazepam and its metabolites in blood and bloodstains. AB - Detection of low levels of flunitrazepam and its metabolites was developed using solid-phase extraction to isolate the drugs from whole liquid blood and dried bloodstains, with subsequent derivatization with pentafluoropropionic anhydride (PFPA) followed by N-(tert-butyldimethyl-silyl)-N-methyl-trifluoroacetamide (MTBSTFA) with 1% TBDMSCI. Analysis was confirmed by gas chromatography/mass spectroscopy using select ion monitoring (sim) in electron impact mode. The limit of detection of this procedure using 1 ml of blood was determined to be 0.1 microgram/dl. PMID- 10371041 TI - Stereoselective analyses of selegiline metabolites: possible urinary markers for selegiline therapy. AB - The stereoselective analysis of selegiline metabolites in human urine and plasma by gas chromatography using the chiral column with the non-chiral reagent was investigated for the differentiation of selegiline therapy from the methamphetamine (MA) abuse. This method gave clear separations of MA and amphetamine (AM) isomers without any artifactual optical-opposite peaks due to the reagent. After the administration of selegiline tablets, desmethylselegiline (DMS), MA and AM were observed as (-)-isomers in the urine and plasma. Within the first 48 h after dosing, approximately 40% of selegiline administered was excreted in urine as these three metabolites. The parent drug, selegiline, was not detected in any urine or plasma samples. On the other hand, MA and AM were observed only as (+)-isomers in the urine of MA abusers. For the distinction of selegiline users from street MA abusers in urinalysis, (-)-DMS, a specific metabolite of selegiline, was not a suitable marker. (-)-DMS rapidly disappeared from urine and was excreted only 1% of the given dose. By the moment analysis with the trapezoidal integration, the mean residence times of (-)-DMS in plasma and urine were 2.7 and 3.8 h, respectively, which were 5-20 times shorter than those of (-)-MA or (-)-AM. The values of AM/MA in the urine increased from 0.24 to 0.67 (r = 0.857) along with time after the selegiline administration. This ratio was not a sufficient marker to differentiate selegiline users from MA abusers, although the values of AM/MA in 74% of MA abusers were less than 0.24. The present GC technique improved the chiral analyses of MA and AM. This chiral analysis is the most useful technique to avoid the misinterpretation in the discrimination between clinical selegiline therapy and illicit MA use. PMID- 10371043 TI - Experimental validation of forensic evidence: a study of the decomposition of buried pigs in a heavy clay soil. AB - In a murder investigation, where the victim had been strangled and buried in a shallow grave, there were discrepancies between the post mortem interval (PMI) as estimated from entomological studies and estimations determined from other evidence. This inconsistency provided the impetus for examining the decay process using pig carcasses as analogues for the human cadaver. The pigs were buried in the immediate vicinity of the original burial site in December 1996, which was the month when the victim was purported to have been interred in the previous year. The buried pigs were then monitored for 5 months which, based on the evidence other than the entomological, was the period over which the corpse was thought to have lain in the ground. The pig corpses were disturbed by scavengers in mid April: this was the same time that the human corpse was discovered in the previous year by scavengers. Insects played no role in the decomposition process until the pig carcasses had been exposed by animals. Blowflies, notably Calliphora vomitoria, were attracted to the exposed tissues and laid eggs from which larvae developed. Calliphora vomitoria is a species often used to estimate PMI. This investigation has shown that soil conditions and low seasonal temperatures had preserved the pig carcasses for longer than might be expected. Using the blowfly larvae to estimate PMI would have produced erroneous results had not the burial environment and exhumation history been investigated. PMID- 10371044 TI - A preliminary study of changes in scalp impedance during the early post-mortem period in rats. AB - An accurate and reproducible technique was used for estimation of scalp impedance (1 kHz) in each of eight rat cadavers, maintained at 9.0 +/- 0.7 degrees C, during the 1 to 144 hour post-mortem period. Scalp impedance increased non linearly, and in a bimodal manner, between 24 and 144 hours post-mortem. The relationship between scalp impedance and post-mortem interval, and the degree of individual variation in the time course of impedance change, were such that scalp impedance is unlikely to be a suitable tool for estimation of time since death. PMID- 10371045 TI - Multivariate analysis ('forensiometrics')--a new tool in forensic medicine. Differentiation between firearm-related homicides and suicides. AB - A forensiometric model to discriminate between homicidal and suicidal firearm fatalities is presented. It is based on a survey of all 45 homicidal and 251 suicidal firearm-fatalities that were examined at the Department of Forensic Medicine in Stockholm, Sweden during the period 1983-92. The model used 15 variables to describe 19.8% of the variation in the data and its goodness of prediction, Q2, was 0.742. The variables ranked in falling order of covariation with homicide (i.e. correlation with or importance in the prediction of homicide) were: presence of other injuries than firearm-wounds, entrance wound in the front part of the chest except precordium, bullet path through clothes, entrance wound in upper extremity, female victim, entrance wound in abdomen, entrance wound in head except temples, central forehead and mouth, and entrance wound in back. The variables most correlated to suicide were: used firearm found close to victim's body, reported suicidal ideation, victim's age, presence of contact wound(s), male victim, presence of farewell letter and entrance wound in mouth. The model was thereafter validated on a test-set of 18 homicides and 84 suicides during 1993-95. All suicides and 16 homicides were classified by the model in agreement with the previous police and forensic medical examinations. Thus, the model's sensitivity to classify homicides is estimated to be 89% and its specificity 100%. PMID- 10371046 TI - A case of fatal intoxication with ammonium sulfate and a toxicological study using rabbits. AB - Agricultural fertilizers such as ammonium sulfate are widely used in house gardens as well as in agriculture, but few case reports or toxicological studies of ingested fertilizers have been reported. This paper investigates a fatal case of ammonium sulfate poisoning and demonstrates its clinical and biochemical findings in rabbits. An 85-year-old woman was found dead lying on the ground outside her house in the middle of March, but the autopsy could not determine the cause of her death. Examination at the police laboratory of the solution in the beer can found next to her showed that it was very likely ammonium sulfate. Our measurement showed a significant increase of ammonium and sulfate ions in serum and gastric contents. The cause of her death was determined as poisoning by ammonium sulfate. The total dose of 1500 mg/kg of ammonium sulfate was administered to three rabbits, all of which showed similar symptoms such as mydriasis, irregular respiratory rhythms, local and general convulsions, until they fell into respiratory failure with cardiac arrest. EEG showed slow, suppressive waves and high-amplitude slowing wave pattern, which is generally observed clinically in hyperammonemia in man and animal. There was a remarkable increase in the concentration of ammonium ion and inorganic sulfate ion in serum, and blood gas analysis showed severe metabolic acidosis. These results, mainly findings by EEG, have shown that a rapid increase in ammonium ions in blood can cause damaging the central nervous system without microscopic change. When the cause of death can not be determined, measurement of ammonium ion, inorganic ion and electrolytes in blood as well as in stomach contents at forensic autopsy is necessary. PMID- 10371047 TI - Fatal neck injuries caused by blank cartridges. AB - We report three cases where fatal neck injuries were caused by blanks from starting pistols. The weapons were loaded with blank cartridges or tear gas cartridges. Neither live ammunition nor any form of projectile was used. All three cases involved a contact discharge. The gas pressure caused by firing the weapons created extensive wound cavities in all three cases. Each victim died from blood loss as a result of ruptured cervical vessels; there were no air embolisms. In one case, a man shot himself eight times with two different starting pistols, and the wounds could be matched to each gun by the muzzle imprint marks on the neck. PMID- 10371048 TI - Solid-phase microextraction for the determination of iodinated trihalomethanes in drinking water. AB - A headspace solid-phase microextraction (HS-SPME) method has been developed for the determination of iodinated trihalomethanes (ITHMs) in treated water samples. Mixed THMs (bromochloroido-, bromodiiodo-, chlorodiiodo-, dibromoiodo- and dichloroiodo-) were previously synthesized since commercial standards are not available. HS-SPME has shorter equilibration times than direct SPME, a cleaner background and a longer fiber life. Experimental parameters such as the selection of SPME coatings, sample volume, extraction time and addition of salts were studied. The Carbowax-divinylbenzene fiber appears to be the most suitable for the determination of ITHMs. Analytical parameters such as linearity, limit of detection and precision were also evaluated. HS-SPME was compared to liquid liquid microextraction for the analyses of spiked treated water samples, obtaining a good agreement. It is concluded that HS-SPME has a great potential for drinking water analysis. PMID- 10371049 TI - Quantitative determination of 2-methoxy-3-isobutylpyrazine in red wines and grapes of Bordeaux using a stable isotope dilution assay. AB - Quantitative analysis of 2-methoxy-3-isobutylpyrazine (MIBP) in grapes and wines was developed, using a stable isotope dilution assay. This was applied to red grapes and wines from the Bordeaux region. The grapes and the wines of the 1995 and 1996 vintages came from the three most frequently used varieties of the region, Merlot, Cabernet Franc and Cabernet Sauvignon. The wines made from Cabernet Sauvignon grapes exhibited levels of MIBP (mean concentration, 12 ng l-1 for 1996 vintage and 13 ng l-1 for 1995 vintage) close to or higher than its odour threshold in wines (10 ng l-1) and slightly higher than the amounts found in the Merlot wines (mean concentration, 8 ng l-1 for 1996 vintage and 4 ng l-1 for 1995 vintage), especially those of the 1996 vintage. The variation in the levels of MIBP in grape samples and in their corresponding wines was monitored at four different stages towards the end of maturation. MIBP was present in all grapes and wines analysed, even in surmaturation. A linear trend was observed between grapes and wines of the three cultivars during maturation. PMID- 10371050 TI - Mercury contamination associated with artisanal gold mining on the island of Mindanao, the Philippines. AB - The Agusan River basin of eastern Mindanao, the Philippines, hosts several centres of artisanal gold mining, the most important of which, Diwalwal, is a significant gold producer in global terms. An investigation of the environmental impacts of artisanal mining in the Agusan system, with particular reference to mercury contamination, was initiated in 1995 following reports of several incidents of human Hg poisoning in the province of Davao del Norte. Results show drainage downstream of Diwalwal is characterised by extremely high levels of Hg both in solution (maximum 2906 micrograms/l) and in bottom sediments (> 20 mg/kg). Filtered surface water Hg levels exceed the WHO Drinking Water guideline and the US-EPA Water Quality Criteria for the Protection of Aquatic Life for a downstream distance of more than 14 km, including channel sections utilised for fishing and potable water supply. The Environment Canada sediment quality Hg Toxic Effect Threshold for the Protection of Aquatic Life is exceeded for a downstream distance of 20 km. Hair Hg data indicate that ballmill and CIP plant operators processing Hg contaminated tailings at eastern Mindanao's principal gold beneficiation centre, Apokon, may be subject to enhanced occupational Hg exposure. It appears that the wider population of this area has not been affected. PMID- 10371051 TI - Transfer of 137Cs and stable Cs from soil to potato in agricultural fields. AB - The concentrations of 137Cs, stable Cs and K were measured in soils and potatoes collected from 26 agricultural fields in Aomori, Japan and soil-to-potato transfer factors of 137Cs and stable Cs were determined. The concentrations of 137Cs derived from fallout deposition and stable Cs in soils were 1-37 Bq kg-1 and 1-11 mg kg-1, respectively. The isotopes, 137Cs and stable Cs, were homogeneously mixed in the rooting zone in the upper 20 cm of soil in agricultural fields. The concentrations of 137Cs and stable Cs in potatoes were 50-2000 mBq kg-1 dry wt. and 0.004-0.13 mg kg-1 dry wt., respectively. The soil to-potato transfer factor of 137Cs was in the range of 0.0037-0.16, which was higher than that of stable Cs of 0.00052-0.080. The transfer factors of 137Cs and stable Cs were correlated and the geometric mean of 137Cs was 0.030, which was four times higher than that of stable Cs at 0.0075. This implied that artificially added 137Cs is more mobile and more easily absorbed by plants than stable Cs in the soil. The concentration of K in potatoes showed a relatively constant value, independent of that in the soil. The transfer factors of both 137Cs and stable Cs decreased with increasing K concentration in the soil, which was mainly supplied as fertilizers to the fields. This suggests that the transfer of both 137Cs and stable Cs from soil to potato was affected by the presence of K in the soil. However, the transfer factors of 137Cs and stable Cs were independent of the amount of organic materials in soils. PMID- 10371052 TI - Long-term fate of organics in waste deposits and its effect on metal release. AB - The long-term chemical evolution in waste deposits and the release of toxic metals was investigated. The degradation of organic matter and hence the potential efflux of heavy metals in a long-term perspective was studied by defining some scenarios for waste deposits containing organic compounds, different longevity and functions of covers and different water and air intrusion rates. The scenarios were based on various transport processes as well as different landfill constructions. The rates of influx of oxygen into both saturated and partially saturated landfills have been estimated. Each scenario takes the form of a mathematical model. The starting point for all the studied cases is the humic phase, i.e. the phase after the methane production has stopped. Based on the different cases studied, it appeared that landfills where the waste is below the water table could have advantages over the other cases. Recognizing that this option is not accepted in most countries we, nevertheless, suggested it should be reevaluated. The main conclusion is that the degradation of humic matter and hence the release of toxic metals can be substantially decreased if potential build-up of hydraulic gradients are avoided and if the landfill is located below the water surface. A conceivable alternative construction would be to place it in a depression--either natural or artificial- and to construct it so that under normal conditions it would always be water saturated. PMID- 10371053 TI - Selenium contents of human milk and infant formulas in Spain. AB - The selenium content of Spanish human milk samples and different milk-based and soy-based infant formulas has been estimated by using a flow injection hydride atomic absorption spectrometric method after microwave digestion of the organic matter. Mean values of 11.4 +/- 3.7 and 10.7 +/- 4.6 ng/ml for colostrum and transitional milk, 8.4 +/- 3.4 and 5.3 +/- 1.9 ng/ml for mature milk at 1 month and up to 2 months respectively, was obtained. These values are close to those reported by others authors in Europe, and lower than the ones from the US, Japan and Korea. Selenium contents of the analyzed infants' formulas ranged from 2.7 to 9.6 ng/ml and from 1.8 to 7.5 ng/ml for soy and milk-based infant formulas, respectively. The variability in selenium contents is large, although mean values are close to the ones given in other European countries. Selenium contents are not usually given on the product. The selenium intakes were estimated assuming that infants fed only human milk. The intakes ranged from 2.0 to 8.4 micrograms/day and from 3.4 to 12.9 micrograms/day for colostrum and transitional milk, respectively, and from 2.6 to 10.3 micrograms/day for mature milk at 1 month, and from 1.2 to 8.3 micrograms/day for milk up to 2 months. The analyzed infant formulas provide significantly less selenium than the 10 micrograms/day corresponding to the recommended daily allowance for infants from 0 to 6 months. PMID- 10371054 TI - Urban anglers' perception of risk from contaminated fish. AB - The Newark Bay Complex includes the Newark Bay, tidal portions of the Hackensack River, Passaic River, Arthur Kill, and Kill van Kull. It is a highly industrialized urban area including five counties and more than 20 local governments with a large racially-mixed population of more than 3 million people. In 1982, research conducted by the New Jersey Department of Environmental Protection (NJDEP) showed elevated levels of chemical contaminants in five species of fish and one type of crab in the Newark Bay Complex. Subsequently, the State of New Jersey adopted advisories to guide citizens on safe consumption practices for fish and crabs. Since then, fish consumption advisories have been issued primarily through the Fish and Game Digest, a publication distributed by the state to licensed anglers. However, anglers in the Complex are not required to have a fishing license because the waters are marine. Therefore, most anglers in this area do not receive advisory information. To gain greater insight into the information sources and risk perceptions of urban anglers, a survey was conducted of 300 anglers at 26 fishing and crabbing locations in the Newark Bay Complex during the summer and early fall of 1995. The objectives of the study were to learn anglers': (1) knowledge of fish consumption advisories; (2) belief in the advisories; (3) perception of how safe fish are to eat; (4) sources for information about fish and fishing; and (5) sources for information on fish consumption advisories. The study concluded that while 60% had heard about advisories, they either did not believe or were unconcerned about health effects from eating contaminated species. In addition, the most used source for information about fish and fishing was other fishermen, while newspapers were selected as a source for information about community news, health, and food safety. PMID- 10371055 TI - Bioavailability of fine dispersed platinum as emitted from automotive catalytic converters: a model study. AB - Automobile exhaust catalytic converters emit fine dispersed elemental platinum, Pt (0), in the nanometer range coated on larger aluminium oxide carrier particles. A pre-requisite for a potential systemic toxic effect of the emitted platinum is its bioavailability which was investigated using laboratory animals. To this end, a model substance was synthesised which consisted of aluminium oxide particles < or = 5 microns onto which platinum particles > or = 4 nm were deposited by a calcination process. These particles closely resemble those emitted from automobile exhaust converters. This model substance was applied to female Lewis rats in two doses by intratracheal instillation; the animals were killed after 1, 7, 28 and 90 days. In addition, the model substance was also applied during a 90-day inhalation study. After microwave digestion of the tissues, the platinum was determined in all organs and body fluids by inductively coupled plasma/mass spectrometry (ICP/MS). Platinum was found in the blood, urine and faeces and all important organs (liver, spleen, kidneys, adrenals, stomach, femur). Based on the platinum content determined in the body fluids and all organs (except the lung and the faeces) it was calculated that up to 16% of the platinum was retained in the lung 1 day after intratracheal instillation and up to 30% of the fine dispersed platinum deposited on an average during 90 days inhalation in the lung was bioavailable. Using size exclusion chromatography (SEC) in combination with ICP/MS, it was shown that > or = 90% of the bioavailable platinum was bound to high molecular weight compounds (approximately 80-800 kDa), most likely proteins. PMID- 10371056 TI - Role of reference materials in analysis of environmental pollutants. AB - This paper discusses the importance and use of reference materials for quality assurance and quality control in environmental analysis. The general classification of reference materials and categorisation of those for chemical composition are presented. The most common reference materials for pollutants in air, water, waste water, soil, sediments, sludge and some biological materials and their producers are tabulated. Definitions, practical recommendations on selection and handling, and application areas of reference materials are also presented. PMID- 10371057 TI - Biochemical indicators of occupational health hazards in Nkalagu cement industry workers, Nigeria. AB - Routine diagnostic tests for some organ-system functions were adopted as biochemical probes in health screening of cement milling workers. Thirty-five volunteers from the Nkalagu cement industry, Nigeria were screened. Out of 14 biochemical parameters studied, raised levels of serum bicarbonates (HCO-), aspartic transaminase (AST) and alanine transaminase (ALT) were observed among cement milling workers compared with a control population. Elevated bicarbonate values were found to differ statistically (P < 0.05) between the quarry workers and the rest of the volunteer work population. In a similar manner, elevated ALT values differ statistically (P < 0.05) between the kilning workers and the rest of the study population. Elevated bicarbonate values are associated with chronic bronchitis, while elevated values of AST and ALT suggest susceptibility to hepatitis, particularly anicteric hepatitis as the elevated values of the transaminase were not matched by a corresponding rise in serum bilirubin. The implications of this study to occupational health screening cannot be overemphasized. PMID- 10371058 TI - Do alterations in the rate of gastric emptying after injection sclerotherapy for oesophageal varices play any role in the development of portal hypertensive gastropathy? AB - Bleeding from portal hypertensive gastropathy (PHG) has been estimated to account for up to 30% of all upper gastrointestinal haemorrhage in patients with cirrhosis and portal hypertension. Although portal hypertension seems to be an essential prerequisite, the precise mechanisms responsible for the development of PHG are unknown. The aim of this study was to examine the role of injection sclerotherapy of oesophageal varices in the development of PHG. Gastric emptying was studied using a radionuclide test meal with the emptying characteristics of a slow liquid in 57 patients with cirrhosis and/or portal hypertension (median age 53 yrs), of whom 34 had received injection sclerotherapy for their oesophageal varices and 20 normal healthy volunteers (median age 42 yrs). As vagal damage is associated with more rapid emptying of liquids, despite hold up of solids, this technique might be expected to demonstrate such damage if gastric emptying was accelerated. The results indicated that there was no difference in the rate of gastric emptying between normal healthy volunteers and portal hypertensive patients. However, patients who had received injection sclerotherapy emptied their stomachs faster than those who had not (p < 0.05). Furthermore, the speed of gastric emptying correlated directly with the number of injections (r = 0.41; p = 0.02) and the volume of sclerosant injected (r = 0.39; p = 0.03). These observations suggest that injection sclerotherapy for oesophageal varices results in disturbances of gastric emptying that may contribute to the pathogenesis of portal hypertensive gastropathy. PMID- 10371059 TI - Reoperations for bleeding portal hypertension. Surgical rescue of surgical failures. AB - BACKGROUND: Surgery for portal hypertension has a low rebleeding rate. Patients that rebleed can be grossly divided into those who die as a consequence of the episode, those who don't die but develop liver failure (remaining as Child-Pugh C) and those who, in spite of the bleeding episode, retain good liver function (Child-Pugh A or B). At our hospital, the latter group is considered for further surgical treatment. We report here the results of surgical rescue of surgical failures. METHODS: In a twenty year period, 36 patients (30 Child-Pugh A, 6 Child Pugh B) were reoperated. The files of these patients were reviewed. RESULTS: Average age was 33 years. Cirrhosis was present in 31 cases. All patients were electively reoperated with portal blood flow preserving procedures. Operative mortality for the whole group was 12% and for the Child-Pugh A group 6.6%. Rebleeding was observed in 5.5%. Postoperative incapacitating encephalopathy was recorded in one case (2.7%). Good quality of life was recorded in 84% of the cases. Survival (Kaplan-Meier) was 78% at 6 months and 69% at 5 years. CONCLUSIONS: Surgical failures in low risk patients (Child-Pugh A or B) can be treated by means of surgery, and a low mortality, re-bleeding and encephalopathy rate can be expected. The performance of a portal blood flow preserving procedure is recommended. PMID- 10371061 TI - Pulmonary lymphangitis carcinomatosa and acute pancreatitis: a rare presentation of choledochal cyst. AB - Pulmonary lymphangitis carcinomatosa is an unusual cause of death in a young adult. This case describes an apparently healthy young woman who presented with severe acute pancreatitis, which is a recognized complication of a choledochal cyst. Autopsy examination revealed advanced malignancy with poorly differentiated adenocarcinoma penetrating the wall of the choledochal cyst and metastatic adenocarcinoma in the lymph nodes, lungs and kidneys. This case emphasises the unusual presentation of a choledochal cyst with acute pancreatitis and the aggressive nature of malignancy associated with this congenital anomaly. PMID- 10371060 TI - Results of surgical treatment (modified Sugiura-Futagawa operation) of portal hypertension associated to complete splenomesoportal thrombosis and cirrhosis. AB - BACKGROUND: Hemorrhagic portal hypertension, secondary to both intrahepatic and extrahepatic portal hypertension, is an uncommon entity. In this condition, the extrahepatic and the intrahepatic obstruction of the portal vein, due to chronic liver disease, produce a more severe form of hemorrhagic portal hypertension that is more difficult to control. The results of surgical treatment (modified Sugiura Futagawa operation) in this subset of patients is analyzed. METHODS: Among 714 patients with a history of hemorrhagic portal hypertension, 14 cases were found with histologically proven liver cirrhosis and complete splenomesoportal thrombosis demonstrated by means of preoperative angiography. Patients with incomplete (partial) splenomesoportal thrombosis were excluded. There were nine males and 5 females with a mean age of 51 years. Alcoholic cirrhosis was demonstrated in 50% of the cases, post hepatitic cirrhosis in 28%, primary biliary cirrhosis in 7%, and cryptogenic cirrhosis in 14%. There were nine Child Pugh A and 5 B cases. All cases were treated by means of our modified Sugiura Futagawa procedure. RESULTS: Bleeding recurrence from esophagogastric varices was shown in one case, colonic varices in one case and hypertensive gastropathy in another of the survivors. Post operative encephalopathy was shown in 3 of the cases. The thirty-six month survival rate was 30% (Kaplan-Meier). CONCLUSIONS: The combination of intrahepatic plus extrahepatic portal hypertension has a worse prognosis. Treatment options are limited (sclerotherapy and/or devascularization), because shunt surgery, TIPS and liver transplantation have a very restricted role and postoperative outcome is poor. PMID- 10371062 TI - Endoscopic management of bleeding ectopic varices with histoacryl. AB - Bleeding from antral and duodenal varices is an uncommon feature in patients with portal hypertension. We report a patient with cirrhosis and portal vein thrombosis, who had a massive bleed from antral and duodenal varices. Bleeding was controlled with endoscopic injection of varices using histoacryl. Endoscopic treatment and the relatively uncommon occurrence of antral and duodenal varices are highlighted. PMID- 10371063 TI - Mucin Hypersecreting Intraductal Papillary Neoplasm of the pancreas. AB - Mucin Hypersecreting Intraductal Papillary Neoplasm is a rare neoplasm that arises from ductal epithelial cells. This entity is distinct from the more commonly known Mucinous Cystadenoma or Mucinous Cystadenocarcinoma. Despite this distinction, it has been erroneously categorized with these more common cystic neoplasms. Characteristic clinical presentation, radiographic, and endoscopic findings help distinguish this neoplasm from the cystadenomas and cystadenocarcinomas. Histopathologic identification is not crucial to the preoperative diagnosis. This neoplasm is considered to represent a premalignant condition and, therefore, surgical resection is warranted. Prognosis, following resection, is felt to be curative for the majority of patients. We present two cases of Mucin Hypersecreting Intraductal Papillary Neoplasm and discuss their diagnosis and surgical therapy. PMID- 10371064 TI - Choledochal cyst associated with polycystic kidney disease: report of a case. AB - We report a very rare case of type I choledochal cyst associated with a polycystic kidney disease. A 48-year-old female had been dependent on hemodialysis for chronic renal failure due to polycystic kidney disease and was incidentally diagnosed to have a dilated common bile duct by an ultrasonography. An endoscopic retrograde cholangiopancreatography showed a spindle-shaped, dilated common bile duct (type I choledochal cyst) without visualization of the pancreatic duct. She underwent a resection of the choledochal cyst. Intraoperative cholangiography showed no reflux of contrast medium into the pancreatic duct. Amylase level of the aspirated bile from the bile duct was not elevated. In the case of choledochal cyst combined with renal fibropolycystic disease, pancreaticobiliary maljunction may not contribute to the etiology of choledochal cyst. In such cases, management of choledochal cyst is still controversial and requires further discussion. PMID- 10371065 TI - Adenoma of the ampulla of Vater: a genetic condition? AB - The etiology of adenoma of the ampulla of Vater is not well understood. Previous authors reported the association of this neoplasm with polycystic kidney disease of two fraternal sisters. They concluded that these two conditions were somehow related. We describe a case of ampullary adenoma associated with polycystic kidney disease. This presentation raises again the question of a possible link between these two diseases. PMID- 10371066 TI - Is banding an acceptable treatment for varices that have not bled (prophylaxis)? PMID- 10371067 TI - Chemoradiation after pancreaticoduodenectomy for pancreatic adenocarcinoma is it of proven benefit? PMID- 10371068 TI - Intractable ascites management: the role of side-to-side portacaval shunt. PMID- 10371069 TI - Does sphincteroplasty predispose to bile duct cancer? PMID- 10371070 TI - Peripheral inflammation is associated with decreased veratridine-induced release of GABA in the rat ventrocaudal periaqueductal gray: microdialysis study. AB - Systemic administration of opiates or direct injection of opioid peptides into the periaqueductal gray (PAG) produces a profound antinociception which is thought to be associated with inhibition of neuronal activity in the PAG. This inhibitory effect has been postulated to result from opiate inhibition of GABAergic neurons in the PAG. Whether this opioid-GABAergic system is affected in acute pain state has not been investigated. The present study was thus designed to determine the effects of unilateral peripheral inflammation on ventrocaudal PAG gamma-aminobutyric acid (GABA) release in the rat using in vivo microdialysis and subsequent high pressure liquid chromatography (HPLC) analysis. Microdialysis was chosen to perform direct and dynamic studies of amino acid concentrations in the PAG in control rats and in animals subjected to acute and prolonged inflammation caused by injection of 120 microl of Complete Freund's Adjuvant (CFA) into the hind paw. GABA release was significantly decreased in the CFA treated groups both 24 h as well as 7 days post-treatment. GABA release decreased to approximately one-fourth that of the 24 h mineral oil control group. Likewise, veratridine-induced release of GABA was decreased in rats treated with CFA 7 days prior to dialysis. Systemic injection of naloxone (5 mg/kg i.p.) caused selective and significant block in the decrease of veratridine-induced release of GABA in the 24 h CFA-treated rats. Taken together with data from our previous studies, these results suggest that the decrease in veratridine-induced GABA release in this study may be due to an increase opiate inhibition of GABA resulting from the induction of acute or prolonged elevation of nociceptive input. PMID- 10371071 TI - Disorders of proprioceptive responses in monkeys after cerebellar lesions: an analysis using the Denny-Brown Collection. AB - We used the Denny-Brown Research Collection to study in detail the reflex responses of monkeys after ablation of the anterior lobe, posterior lobe or the entire cerebellum. The Collection includes written, film and histological records, and photographs of the brain at autopsy. Large cerebellar ablations severely suppress proprioceptive responses, thereby significantly impairing the capacity to stand, walk, and hop. Cutaneous reflexes are also impaired, although more selectively, permitting expression of normally suppressed responses such as magnet reactions and tactile avoiding responses. Enhancement (release) of responses to truncal cutaneous stimulation, along with suppression of opposing proprioceptive responses, leads to postures of persistent flexion. Large cerebellar lesions also interfere with reflex responses mediated by visual and vestibular systems. More limited cerebellar ablations have similar, but less severe effects. PMID- 10371072 TI - Parieto-occipital glucose hypometabolism in Parkinson's disease with autonomic failure. AB - To investigate the characteristics of regional cerebral metabolism in a subgroup of patients with Parkinson's disease and autonomic failure, we studied seven patients with Parkinson's disease with autonomic failure (PA group), 11 patients with Parkinson's disease without apparent autonomic failure (PD group), and nine normal controls using fluoro-deoxyglucose positron emission tomography (FDG-PET). To determine differences in metabolic distribution among these groups, regional relative glucose metabolic rates (RGMR), which were normalized with cerebellar values, were calculated and age-adjusted covariance analyses were done. When compared with that of controls. RGMR in the cerebral cortex of the PA group was markedly reduced in the occipital cortex (P<0.001), inferior parietal cortex (P<0.005) and superior parietal cortex (P<0.005), but without a decrease in the sensory motor and medial temporal cortices, putamen and thalamus. In contrast, the PD group did not show significant focal hypometabolic distribution. Our findings raise the possibility that Parkinson's disease with autonomic failure may overlap with the features of dementia with Lewy bodies. PMID- 10371073 TI - Malignant brain tumour mortality among children and adolescents: geographical distribution in Spain. AB - Log-linear Poisson mixed models were used to study provincial malignant brain tumour (MBT) mortality among children and adolescents in Spain (1975-1992) in order to investigate the influence of specific socio-economic factors and to produce smoothed estimators of standardised mortality ratios (SMRs). Interdependence between geographical units was taken into account by including provinces as random effect terms nested with the corresponding Autonomous Region (Spain's administrative divisions). MBT mortality showed a positive association with three variables: non-cancer-related infant mortality; percentage of provincial land surface area devoted to agriculture; and industrial/construction activity. According to the final model, SMRs increased by 4% with every 10% rise in the area devoted to agriculture, with a 4% excess risk predicted for every increase in non-cancer-related infant mortality of 1 per 1000 person-years, and a 7% excess risk for contiguous categories of industrial/construction activity. By smoothing extreme values caused by random variability, the regression model yielded a reasonable estimation of SMRs. While infant mortality may be linked to the quality of medical care available, the relationship seen between MBT mortality and agricultural area accords with the excess risk reported for farmers' offspring. Finally, industrial activity might be regarded as a risk factor or as a marker of other conditions also associated with these tumours. PMID- 10371074 TI - Activated protein C resistance in young African American patients with ischemic stroke. AB - BACKGROUND: It has been proposed that activated protein C resistance (APCR) due to the factor V Leiden (FVL) mutation may be a risk factor for stroke in young adults. However, this may not be the case for all ethnic groups due to variability in the prevalence of the FVL mutation. METHODS: Case series from a university neurology clinic. Patients with an APCR ratio of 2.2 or below were tested for the FVL mutation (nine patients). Patients on warfarin were also tested for the FVL mutation (14 patients). RESULTS: 38 African American patients under age 55 with an arterial stroke were identified. The mean age of the patients is 43.1 years. Five percent had an APCR ratio of 2.0 or below. None of the patients with an APCR ratio of 2.2 or below or the patients directly tested for the FVL mutation had the mutant allele. CONCLUSIONS: Activated protein C resistance due to the FVL mutation does not appear to be a major risk factor for stroke in young African Americans. Other, as yet unidentified, mechanisms leading to activated protein C resistance may be important in a small subset of young African American stroke patients. PMID- 10371075 TI - Secondary reduction of alpha7B integrin in laminin alpha2 deficient congenital muscular dystrophy supports an additional transmembrane link in skeletal muscle. AB - The integrins are a large family of heterodimeric transmembrane cellular receptors which mediate the association between the extracellular matrix (ECM) and cytoskeletal proteins. The alpha7beta1 integrin is a major laminin binding integrin in skeletal and cardiac muscle and is thought to be involved in myogenic differentiation and migration processes. The main binding partners of the alpha7 integrin are laminin-1 (alpha1-beta1-gamma1), laminin-2 (alpha2-beta1-gamma1) and laminin-4 (alpha2-beta2-gamma1). Targeted deletion of the gene for the alpha7 integrin subunit (ITGA7) in mice leads to a novel form of muscular dystrophy. In the present study we have investigated the expression of two alternative splice variants, the alpha7B and beta1D integrin subunits, in normal human skeletal muscle, as well as in various forms of muscular dystrophy. In normal human skeletal muscle the expression of the alpha7 integrin subunit appeared to be developmentally regulated: it was first detected at 2 years of age. In contrast, the beta1D integrin could be detected in immature and mature muscle in the sarcolemma of normal fetal skeletal muscle at 18 weeks gestation. The expression of alpha7B integrin was significantly reduced at the sarcolemma in six patients with laminin alpha2 chain deficient congenital muscular dystrophy (CMD) (age >2 years). However, this reduction was not correlated with the amount of laminin alpha2 chain expressed. In contrast, the expression of the laminin alpha2 chain was not altered in the skeletal muscle of the alpha7 knock-out mice. These data argue in favor that there is not a tight correlation between the expression of the alpha7 integrin subunit and that of the laminin alpha2 chain in either human or murine dystrophic muscle. Interestingly, in dystrophinopathies (Duchenne and Becker muscular dystrophy; DMD/BMD) expression of alpha7B was upregulated irrespective of the level of dystrophin expression as shown by a strong sarcolemmal staining pattern even in young boys (age <2 years). The expression of the beta1D integrin subunit was not altered in any of our patients with different types of muscular dystrophy. In contrast, sarcolemmal expression of beta1D integrin was significantly reduced in the alpha7 integrin knock-out mice, whereas the expression of the components of the DGC was not altered. The secondary loss of alpha7B in laminin alpha2 chain deficiency defines a biochemical change in the composition of the plasma membrane resulting from a primary protein deficiency in the basal lamina. These findings, in addition to the occurrence of a muscular dystrophy in alpha7 deficient mice, implies that the alpha7B integrin is an important laminin receptor within the plasma membrane which plays a significant role in skeletal muscle function and stability. PMID- 10371077 TI - Cytotoxic T lymphocyte-mediated cell death in paraneoplastic sensory neuronopathy with anti-Hu antibody. AB - Paraneoplastic sensory neuronopathy (PSN) has been shown to harbor characteristic anti-neuronal autoantibody 'anti-Hu' in their sera and cerebrospinal fluid. Creation of animal models exhibiting clinical or pathological features seen in PSN by means of passive transfer of anti-Hu positive IgG has not been achieved. Although, anti-Hu antibody was shown to induce neuronal cell lysis in vitro, this result has not been reproduced so far. Since prominent T cell infiltration are seen in the central nervous system and posterior spinal ganglion of the patients with anti-Hu syndrome, we studied cytotoxic T cell (CTL) activity in peripheral mononuclear cells from a patient with PSN harboring anti-Hu antibody. The activated CD8+ T cells from the patient's venous blood were shown to lyse her own fibroblasts which were incubated with interferon-gamma to induce HLA class I molecules on their surface and the recombinant HuD protein was injected into the cells by microinjector. This is the first report showing the existence of CTL in a patient with PSN. PMID- 10371076 TI - Vasomotor response to CO2 and L-Arginine in patients with severe internal carotid artery stenosis; pre- and post-surgical evaluation with transcranial Doppler. AB - BACKGROUND AND AIM: Carotid artery disease may cause both thromboembolism and cerebral blood flow disturbances, particularly in subjects with impaired hemodynamic compensatory mechanisms. The aim of this study was to evaluate by transcranial Doppler (TCD) the hemodynamic changes induced by CO2 and L-Arginine stimulation in a selected population with severe unilateral carotid stenosis (70 80%), before and after carotid endarterectomy, in order to determine the effect of surgery in the vascular hemodynamics of these patients. METHODS: We studied 20 subjects (mean age 66.4 years) consecutively admitted to our institute with ischemia and unilateral severe internal carotid artery stenosis (70-80%) detected by Color Doppler. All patients underwent arterial digital subtraction angiography to confirm the ultrasonographic evaluation. TCD was performed bilaterally; blood flow velocity was monitored during CO2 and L-Arginine stimulation both in basal conditions and three months after surgery. RESULTS: After endarterectomy, mean velocity increased in response to both stimuli with a trend toward statistical significance. A significantly lower reactivity to L-Arginine on the stenotic side was found in the pre-operative phase: this asymmetrical reactivity was no longer observable after carotid endarterectomy. CONCLUSIONS: We found a statistically significant difference in L-Arginine reactivity in the stenotic side of patients with severe unilateral internal carotid stenosis. This is probably related to an alteration of the endothelium function due to the carotid pathology, since the abnormalities disappeared three months after endarterectomy. PMID- 10371078 TI - White matter dementia in CADASIL. AB - Cerebral white matter disorders may be associated with profound neurobehavioral dysfunction. We report a 62-year-old man who had a slowly progressive 25-year history of personality change, psychosis, mood disorder, and dementia. Neurologic examination disclosed abulia, impaired memory retrieval, and preserved language, with only minimal motor impairment. Neuropsychological testing found a sustained attention deficit, cognitive slowing, impaired learning with intact recognition, and perseveration. Magnetic resonance imaging of the brain revealed extensive leukoencephalopathy. Right frontal brain biopsy showed ill-defined white matter pallor with hyaline narrowing of white matter arterioles. Granular osmiophilic material adjacent to vascular smooth muscle cells on electron microscopy of a skin biopsy, and an arginine for cysteine replacement at position 169 in the 4 EGF motif of the notch 3 region on chromosome 19q12 established the diagnosis of cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL). This case illustrates that CADASIL can manifest as an isolated neurobehavioral disorder over an extended time period. The dementia associated with CADASIL closely resembles that which may occur with other white matter disorders, and represents an example of white matter dementia. PMID- 10371079 TI - The mitochondrial DNA A3243G mutation in Portugal: clinical and molecular studies in 5 families. AB - Out of 90 Portuguese patients with mitochondrial cytopathy, six harbored the A3243G mutation in the mtDNA tRNA(Leu(UUR)) gene ('MELAS mutation'). They had heterogeneous clinical features, including myopathy with stroke-like episodes, progressive external ophthalmoparesis, diabetes mellitus, and subacute encephalopathy. Histochemical and biochemical analyses of muscle biopsies showed abundant ragged-red fibers reacting positively with the cytochrome oxidase stain, and decreased respiratory chain enzyme activities. On average, the proportion of mutated mtDNA was 67% (20-88%) in tissues from patients and 21% (0-49%) in blood from 20 maternal relatives. The proportion of mutated mitochondrial genomes in muscle did not correlate with clinical presentation or duration of disease. This study, the first in Portuguese patients, confirms the frequent occurrence of the A3243G mutation in patients with mitochondrial diseases, and emphasises the usefulness of genetic testing in reaching a correct diagnosis. PMID- 10371080 TI - Late appearance of acanthocytes during the course of chorea-acanthocytosis. AB - A case of chorea-acanthocytosis (CA) syndrome is described. The presence of acanthocytes has usually been considered an important diagnostic marker of CA. However, it is not specific and other neurological diseases have to be considered. In the present report we rule out other diagnostic possibilities and show that the acanthocytes in the peripheral blood smears can appear even later during the course of the disease. PMID- 10371081 TI - Incongruous homonymous hemianopic scotoma. AB - We report a patient presenting with incongruous homonymous hemianopic scotoma due to infarction in the territory of the lateral posterior choroidal artery. Imaging studies showed that the patient had a fresh infarct in the lateral geniculate body causing this unusual visual field defect. PMID- 10371082 TI - Neurodegeneration in Xeroderma Pigmentosum: a trinucleotide repeat mutation analysis. AB - Xeroderma Pigmentosum (XP) is a rare autosomal recessive disorder caused by defects in DNA repair. In some forms, it is clinically and pathologically characterized by neurological involvement and premature neuronal death. This study explores the hypothesis that defects in DNA repair in XP may contribute to neurological involvement by destabilizing trinucleotide repeats during replication causing expansion mutations into disease producing ranges. Trinucleotide repeat instability in each of the genes causing Machado-Joseph Disease, myotonic dystrophy, Kennedy's Disease and Huntington's Disease was analyzed by performing single genome PCR. The results of trinucleotide repeat analysis of 360 single genomes from three different forms of XP showed that the size of the repeats were in the normal range and that there was no mitotic instability. These results suggest that in XP, trinucleotide repeat expansion mutations are not involved in the pathophysiology of neurodegeneration. PMID- 10371083 TI - Histiocytic necrotizing lymphadenitis (Kikuchi's disease) with aseptic meningitis. AB - Histiocytic necrotizing lymphadenitis, or Kikuchi's disease (KD), is a self limited clinicopathologic entity recognized increasingly worldwide. A 27-year-old man with cervical lymphadenopathy and fever who was diagnosed with KD developed mild headache with no nuchal rigidity. The cerebrospinal fluid (CSF) was sterile and contained 78 white blood cells/mm3 with lymphocytes predominating, accompanied by smaller numbers of monocytes and granulocytes. This abnormality normalized spontaneously over 5 weeks. Eleven similar cases have been reported, all but one from Japan. The development of meningitis in KD was observed in four (9.8%) of 41 KD patients we have treated, suggesting that the meningitis was related to KD and not merely coincidental. PMID- 10371084 TI - Wernicke encephalopathy-like symptoms as an early manifestation of Creutzfeldt Jakob disease in a chronic alcoholic. AB - A case of Creutzfeldt-Jakob disease (CJD) with presenting Wernicke encephalopathy (WE)-like symptoms and severe insomnia is presented. An 80-year-old alcoholic man with a 6 month history of tremors, ataxia, memory loss and confabulation, developed profound insomnia, confusion, and delirium with vivid hallucinations. Polysomnography revealed a marked reduction of sleep time, with central-type sleep apnea. Neither myoclonus nor periodic synchronous discharge (PSD) was observed. An autopsy revealed diffuse spongiform changes and astrocytosis throughout the cerebral gray matter, with severe involvement of the mammillary bodies and thalamus. Prion protein (PrP) immunostaining was positive in kuru plaques in the cerebellum, PrP polymorphism at codon 129 was heterozygous Met/Val, and proteinase K resistant PrP (PrP(res)) was demonstrated by Western blotting. The lack of necrotizing lesions in the mammillary bodies, thalamus, and periaqueductal gray matter could rule out WE. The data suggest that the present case of CJD is consistent with PrP(res) type 2 (CJD M/V 2), but was unique in the lack of some typical CJD signs and the presence of signs of WE and sleep abnormalities. PMID- 10371085 TI - Actions of benzodiazepines and the benzodiazepine antagonist flumazenil may involve adenosine. AB - It has recently been reported that the benzodiazepine antagonist flumazenil (formerly Ro15-1788) has depressant actions of its own on hippocampal potentials. It is noted here that similar results were obtained over ten years earlier, in association with work showing that benzodiazepine agonists and antagonists could inhibit the uptake of adenosine. Such an action would lead to the extracellular accumulation of adenosine which, because of its ability to suppress transmitter release, could explain the recent results. PMID- 10371086 TI - Hashimoto's encephalopathy. PMID- 10371087 TI - Role of IgE in the development of allergic airway inflammation and airway hyperresponsiveness--a murine model. AB - The importance of IgE in airway inflammation and development of AHR in allergen sensitized mice has been compared and contrasted in different models of sensitization and challenge. Using different modes of sensitization in normal and genetically manipulated mice after anti-IgE treatment, we have been able to distinguish the role of IgE under these different conditions. Striking differences in the three sensitization protocols were delineated in terms of the role of allergen-specific IgE, extent of eosinophilic airway inflammation, and development of AHR (Table 1). The highest levels of IgE and eosinophil infiltration (approximately 20-fold increases) were achieved after systemic sensitization with allergen (plus adjuvant) followed by repeated airway challenge. Passive sensitization with allergen-specific IgE followed by limited airway challenge induced a modest eosinophilic inflammatory response in the airways despite high levels of serum IgE. Exposure to allergen exclusively via the airways also resulted in a modest serum IgE response and a limited eosinophilic inflammatory response (approximately fourfold increases). Under all of these conditions, inhibition of IL-5-mediated eosinophilic airway inflammation was associated with attenuation of AHR. In contrast, the differences in the responses to the different modes of allergen exposure were associated with differences in the requirements for IgE in the development of AHR (Table 1). In the two models associated with mild eosinophil infiltration (passive sensitization and exclusive airway exposure), IgE was required for the development of AHR but did not substantially enhance airway inflammation on its own. However, IgE-allergen interaction was able to enhance T-cell function in vitro and induce T-cell expansion in vivo. In mice systemically sensitized and challenged via the airways, IgE (or IgE-mediated mast-cell activation) was not required for T-cell activation, eosinophilic inflammation and activation in the airways, or development of AHR. This was most clearly seen in B-cell-deficient and mast-cell-deficient, low-IgE-responder mouse strains (B6, B10) and in anti IgE-treated high-IgEresponder mice (BALB/c). At the same time, we confirmed the importance of IgE in the induction of immediate-type hypersensitivity (mast-cell activation, immediate cutaneous hypersensitivity, passive cutaneous and systemic anaphylaxis). These differences were also highlighted by the means used to detect altered airway function. Passive sensitization and limited airway challenge or exclusive airway exposure to allergen over 10 days elicited changes in airway function that could be detected only in tracheal smooth-muscle preparations exposed to EFS. In contrast, systemic sensitization followed by repeated airway challenge resulted not only in changes in the contractile response to EFS but also in increased responsiveness to inhaled MCh. Thus, these results distinguish not only the differential involvement of IgE and eosinophil numbers but also their contribution to the readouts used to monitor airway function. Based on these studies, we conclude that IgE plays an important role in the development of airway inflammation and AHR under conditions in which limited IL-5-mediated eosinophilic airway infiltration is induced. In conditions where a robust eosinophilic inflammation of the airways is elicited, IgE (and IgE-mediated mast cell activation) does not appear to be essential for airway inflammation and the development of AHR, detected as increased responsiveness to inhaled MCh. These findings reveal the potential importance of differential targeting in the treatment of allergic diseases with a predominance of IgE-mediated symptoms, e.g., allergic rhinitis and conjunctivitis, where anti-IgE may be an effective therapy, compared to those diseases with a predominant inflammatory component, e.g., AHR in atopic bronchial asthma, where anti-inflammatory or anti-IL-5 therapy may be more beneficial. PMID- 10371088 TI - Mast-cell growth and differentiation. PMID- 10371089 TI - Long-term treatment with allergoid immunotherapy with Parietaria. Clinical and immunologic effects in a randomized, controlled trial. AB - BACKGROUND: Specific immunotherapy (SIT) is a valuable treatment for respiratory allergy, and the use of chemically modified allergens (allergoids) has improved its safety, as testified by several studies. We evaluated the effects of a SIT course with an allergoid extract of Parietaria pollen in a double-blind, placebo controlled trial. METHODS: The study was double-blind in the first year; then it was prolonged up to 3 years with all patients on active treatment. Clinical effectiveness, safety, skin reactivity, systemic immunologic parameters, and subjective assessment were evaluated. We also had available a self-evaluation recorded in a follow-up visit 4 years after the discontinuation of SIT. RESULTS: A significant reduction of the symptoms plus drug intake scores during the pollen seasons was observed in the patients receiving active SIT. The placebo patients, after switching to active SIT, also showed significant clinical improvement. The clinical efficacy persisted during years 2 and 3 of treatment. After year 1, the actively treated patients reported a significant subjective improvement (frequency of symptoms, P = 0.001; duration of symptoms, P = 0.024; physical performance, P = 0.043) compared with the placebo group. The self-evaluation by visual analog scale showed that all patients maintained a significant clinical improvement up to 4 years after discontinuing SIT (year 1: active=+31.6%, placebo=-15.7%; year 7: active=+35.8%, placebo=+31.3%). The systemic immunologic changes after active SIT paralleled those described elsewhere (IgE decreased from 22 to 9 and from 21 to 8 IU/ml; IgG4 increased from 43 to 87 and from 18 to 60 IU/ml). A significant decrease in skin reactivity to three different allergen concentrations was observed at year 3 compared with pretreatment values (P<0.05). CONCLUSIONS: The investigational SIT with Parietaria appeared to be effective and safe; a 3-year course of treatment achieved a long-lasting efficacy. PMID- 10371091 TI - Increased risk of allergy in children due to formaldehyde exposure in homes. AB - BACKGROUND: Formaldehyde levels were measured in 80 houses in the Latrobe Valley, Victoria, Australia. An association between exposure to formaldehyde and sensitization to common aeroallergens has been suggested from animal trials, but no epidemiologic studies have tested this hypothesis. METHODS: A total of 148 children 7-14 years of age were included in the study, 53 of whom were asthmatic. Formaldehyde measurements were performed on four occasions between March 1994 and February 1995 with passive samplers. A respiratory questionnaire was completed, and skin prick tests were performed. RESULTS: The median indoor formaldehyde level was 15.8 microg/ m3(12.6ppb), with a maximum of 139 microg/m3 (111 ppb). There was an association between formaldehyde exposure and atopy, and the adjusted odds ratio was 1.40 (0.98-2.00, 95% CI) with an increase in bedroom formaldehyde levels of 10 microg/m3. Furthermore, more severe allergic sensitization was demonstrated with increasing formaldehyde exposure. On the other hand, there was no significant increase in the adjusted risk of asthma or respiratory symptoms with formaldehyde exposure. However, among children suffering from respiratory symptoms, more frequent symptoms were noted in those exposed to higher formaldehyde levels. CONCLUSIONS: Low-level exposure to indoor formaldehyde may increase the risk of allergic sensitization to common aeroallergens in children. PMID- 10371090 TI - Association of ambient air-pollution levels with acute asthma exacerbation among children in Singapore. AB - BACKGROUND: Air-pollution levels have been shown to be associated with increased morbidity of respiratory diseases. METHODS: Data for ambient air-pollutant levels, meteorologic factors, and hospitalization or emergency room (ER) visits for acute asthma in Singapore children over a 5-year period (1990-4) were obtained and analyzed for associations by time-series methods. RESULTS: Throughout this period, the annual mean and 24-h mean levels for sulfur dioxide (SO2), nitrogen dioxide (NO2), and total suspended particles (TSP) and maximum 1 h daily average for ozone were generally within the air-quality guidelines established by the World Health Organization (WHO). However, positive correlation between levels of each of these pollutants and daily ER visits for asthma was observed in children aged 3-12 years, but not among adolescents and young adults (13-21 years old). The association with SO2 and TSP persisted after standardization for meteorologic and temporal variables. An adjusted increase in 2.9 ER visits for every 20 microg/m3 increase in atmospheric SO2 levels, lagged by 1 day, was observed on days when levels were above 68 microg/m3. With TSP, an adjusted increase of 5.80 ER visits for every 20 microg/m3 increase in its daily atmospheric levels, lagged by 1 day, was observed on days with levels above 73 microg/m3. Similar results were also obtained after controlling for autocorrelation by time-series analysis. CONCLUSIONS: These associations were observed even though the overall levels of all pollutants were generally within the air-quality guidelines established by the WHO. These findings suggest that asthmatic children are susceptible to increased levels of air pollutants, particularly SO2 and TSP, although the ambient levels are generally within "acceptable" ranges. PMID- 10371092 TI - Cellular infiltration and cytokine mRNA expression in perennial allergic rhinitis. AB - BACKGROUND: Allergen challenge in allergic rhinitis patients leads to local eosinophilia and Th2-type cytokine expression. Natural exposure to grass pollen is additionally characterized by epithelial mast-cell infiltration. We hypothesized that perennial allergic rhinitis is also associated with T-cell and eosinophil infiltration of the nasal mucosa, local Th2-type cytokine expression, and increased numbers of nasal epithelial mast cells. METHODS: Nasal biopsies from perennial allergic rhinitis patients and controls were analysed by immunocytochemistry for different cell populations and in situ hybridization for cytokine mRNA-expressing cells. RESULTS: Perennial allergic rhinitis was associated with increased numbers of submucosal CD3+ T cells (P=0.05), EG2+ activated eosinophils (P=0.01), and CD68+ macrophages (P=0.01) compared to controls. Epithelial, but not submucosal, tryptase-positive mast cells were also elevated in rhinitics compared to controls (P=0.01). The numbers of cells expressing interleukin (IL)-5 were higher (P=0.01) and the numbers of cells expressing IL-2 were lower (P=0.04) in rhinitic patients than controls. There were no significant differences for either IL-4 or interferon-gamma between the groups. CONCLUSIONS: Perennial allergic rhinitis is characterized by mast-cell migration into the epithelium; submucosal infiltration by T cells, eosinophils, and macrophages; and an imbalance in local T-cell cytokine production in favour of enhanced IL-5 and reduced IL-2 expression. PMID- 10371093 TI - Cow's milk allergy: diagnostic accuracy of skin prick and patch tests and specific IgE. AB - BACKGROUND: The objective of the present study was to evaluate the relevance of skin tests and the concentration of cow's milk-specific IgE antibodies in correlation with oral cow's milk challenge in infants with suspected cow's milk allergy. METHODS: The study material comprised 143 infants under the age of 2 years who had undergone a diagnostic elimination challenge because of suspected cow's milk allergy in 1996. Cow's milk-specific IgE was measured, and skin prick and patch tests were performed. RESULTS: Of the 143 oral cow's milk challenges performed, 72 (50%) were positive. Of the positive reactions, 22 involved immediate-type reactions. In 50 patients, delayed-onset reactions of eczematous or gastrointestinal type appeared. Of the infants with challenge-proven cow's milk allergy, 26% showed elevated IgE concentrations to cow's milk, 14% had a positive skin prick test, and 44% had a positive patch test for cow's milk. Interestingly, in most patch test-positive patients, the prick test for cow's milk was negative. CONCLUSIONS: Our study demonstrated that many patients with a negative prick test result had a positive patch test to cow's milk. The patch test was a more sensitive method than the prick test or RAST to detect cow's milk allergy in this study population. Our results indicate that patch testing will significantly increase the probability of early detection of cow's milk allergy. Confirmation of the diagnosis is essential in patients with negative test results but a clinical suspicion of food allergy, and in patch test-positive patients. For this purpose, the most reliable method is the elimination-challenge procedure. PMID- 10371094 TI - Cow's milk IgE and IgG antibody responses to cow's milk formulas. PMID- 10371095 TI - Continuous antihistamine treatment controls allergic inflammation and reduces respiratory morbidity in children with mite allergy. AB - BACKGROUND: Allergic reaction is characterized by a complex inflammatory process. Some of the new antihistamines have antiallergic effects and can affect the inflammatory cell recruitment via adhesion molecule downregulation. We aimed to assess in a 12-month study whether continuous treatment with an antihistamine (terfenadine) can reduce respiratory symptoms and local inflammation in children with mite allergy. METHODS: The study was double-blind and placebo-controlled: it involved two parallel groups of children suffering from rhinoconjunctivitis and/or mild intermittent asthma due to mite allergy. They received either terfenadine (1 mg/kg per body weight per day) or placebo for 1 year. Nasal, conjunctival, and bronchial symptoms were recorded by diary cards; at each of the programmed control visits, a nasal scraping for inflammatory cells and ICAM-1 was performed. Some additional clinical parameters were also recorded: days of school absence, extra visits for acute respiratory symptoms, and days of hospital admission. RESULTS: Only children treated with terfenadine achieved significant control of symptoms (P<0.05 in 8 out of 12 months) and allergic inflammation, as shown by inflammatory cell infiltrate and ICAM-1 expression at nasal level (P<0.001), and had significantly fewer extra visits and school absences than the placebo group (P<0.03). No side-effects were reported in either group. CONCLUSIONS: The present study demonstrates that continuous terfenadine treatment (1 mg/kg body weight per day) could decrease respiratory symptoms and allergic inflammation, and it had an additional antiallergic effect in reducing ICAM-1 expression on nasal epithelial cells. Therefore, the present results confirm the efficacy of a long-term therapeutic strategy in controlling allergic inflammation. PMID- 10371096 TI - Bronchial allergen challenge in subjects with low levels of allergic sensitization to indoor allergens. AB - BACKGROUND: Low skin reactivity to common inhalant allergens is frequently found in asymptomatic individuals as well as in patients with respiratory complaints. However, most studies on bronchial allergen challenge concern patients with high levels of allergic sensitization. The present study was directed to bronchial reactions after allergen challenge in subjects with low skin reactivity to Dermatophagoides pteronyssinus or cat dander. METHODS: Titrated intracutaneous skin tests, skin prick tests, specific IgE assays, histamine release on washed leukocytes, and bronchial histamine and allergen-challenge tests were performed in 20 subjects with an intracutaneous skin test threshold for cat dander (Felis domesticus) or D. pteronyssinus above 0.1 BU/ml (mean wheal diameter in skin prick test with 10000 BU/ml: 4.4mm). Ten of the 20 patients had specific IgE below the detection limit in at least one of the three IgE assays which were done. Fifteen patients had a specific IgE level below 2 kU/I in all three tests. As a positive control group, the same parameters were studied in seven moderately sensitized patients with an intracutaneous skin test threshold below 0.1 BU/ml (mean wheal diameter with 10000 BU/ml: 7.2mm). RESULTS: The 20 subjects with low levels of allergic sensitization had an early decrease in FEV1 of 8.6% (P<0.01) and a mean late decrease of 6.3% (P<0.05). There was a trend for decrease in PC20 histamine 24h after allergen challenge (-0.4 doubling doses, P=0.09). CONCLUSIONS: In this group of subjects with low levels of allergic sensitization, a statistically significant early and late decrease in FEV1 was found. However, the decrease in lung function was small and unnoticed by most patients. The increase in nonspecific bronchial hyperresponsiveness after bronchial allergen challenge did not reach statistical significance in the study group. The results indicate that allergen exposure in patients with low levels of allergic sensitization may lead to airways changes in the absence of acute symptoms. PMID- 10371097 TI - Turbutest in the training of asthmatic Turbuhaler users. AB - BACKGROUND: Correct utilization of inhalation devices is a key factor in asthma management. Objective assessment of the ability to use inhaler devices is therefore fundamental. METHODS: The objective was to assess objectively the inhalation technique of Turbuhaler users who reported having no difficulty in using such a device. A total of 600 asthmatic patients of our allergology outpatient department, daily users of Turbuhaler devices, were evaluated with Turbutest. This is a simple portable apparatus which connects a Turbuhaler inhaler with an electronic device to allow semiquantitative assessment of inhalation technique, with a score ranging from 0 (worst) to 3 (best), readily visualized on the apparatus. Patients were allowed to practice with Turbutest, assisted by an allergologist, until they improved their scores. A second Turbutest assessment was performed at a 2-month follow-up visit. RESULTS: On the first assessment, one-third of the patients had scores inadequate to allow a good inhalation, and only 20% achieved a score of 3. On the second assessment, significantly higher Turbutest scores were observed. CONCLUSIONS: Turbutest is a valuable tool in asthmatic patients' training, allowing identification and improvement of an inadequate inhalation technique with Turbuhaler. PMID- 10371098 TI - Perceived prevalence of peanut allergy in Great Britain and its association with other atopic conditions and with peanut allergy in other household members. AB - BACKGROUND: Despite increasing awareness of peanut allergy, little is known of its prevalence. We report on a two-stage interview survey conducted in Great Britain. METHODS: A total of 16434 adults (aged 15+ years) reported their own allergies and atopies and named cohabitants with peanut allergy (stage 1). Follow up interviews were conducted with identified sufferers from peanut allergy (stage 2). RESULTS: At stage 1, peanut allergy was reported in 58 respondents and 205 other household members. When we accounted for cases where peanut allergy was unconfirmed or newly reported at stage 2, the prevalence, based on 124 confirmed sufferers, was estimated as 0.48% (95% confidence interval 0.40%-0.55%). The prevalence in children (0.61%, 0.41%-0.82%) was slightly higher than in adults; age-of-onset was strikingly earlier. Prevalence was strongly associated with other atopies, particularly tree-nut allergy. Cases tended significantly to cluster in households. Half of cases had never consulted a doctor. Exactly 7.4% reported being hospitalized after a reaction. CONCLUSIONS: Peanut allergy is reported by 1 in 200 of the population and is commoner in those reporting other atopies. The fact of similar rates in children and adults argues against a recent marked rise in prevalence. The frequency and potential lethality of this disorder emphasize the need for sufferers to demographic factors, other food allergies, atopic conditions, and allergy in family/household members. Our study comprised a screening survey and detailed interviews with sufferers identified. The frequency and potential lethality of this disorder emphasize the need for sufferers to receive correct medical advice on management [corrected]. PMID- 10371099 TI - Interleukin-8 inhalation directly provokes bronchoconstriction in guinea pigs. AB - BACKGROUND: Although it has been reported that the concentration of interleukin (IL)-8 in nasal lavage fluid and sputum and its production in bronchial epithelium were increased in asthmatic subjects, the direct effects of IL-8 on the airways in vivo is unclear. METHODS: We examined bronchoconstriction in response to IL-8 inhalation through an endotracheal cannula in anesthetized, artificially ventilated guinea pigs. RESULTS: Inhalation of IL-8 at concentrations of 1 and 10 microg/ml caused significant bronchoconstriction, as revealed by the elevation of pressure at the airway opening. Moreover, the bronchoconstriction induced by IL-8 was significantly inhibited by the antihistamines diphenhydramine and terfenadine, suggesting the involvement of histamine release in the IL-8-induced bronchoconstriction. No significant leukocyte infiltration was observed in the bronchoalveolar lavage fluid or histologic findings 25 min after the first IL-8 inhalation. CONCLUSIONS: IL-8 provokes bronchoconstriction without leukocyte accumulation in the airways, mediated in part by histamine release, in guinea pigs. PMID- 10371100 TI - Biologic activity of Dermatophagoides siboney and Blomia tropicalis allergens in exposed and unexposed mite-allergic individuals. Effect of patient selection on the biologic standardization of mite extracts. AB - BACKGROUND: This study aimed to investigate the influence of patient selection criteria, i.e., mite-allergic individuals exposed and not exposed to Dermatophagoides siboney and Blomia tropicalis, on the biologic activity of mite extracts. Determination of the potency of mite extracts in vivo requires selection of patients with a clinical history of mite allergy. In Scandinavia, there are some anamnestic criteria for mite allergy, whereas in the tropics, where patients are continuously exposed to high levels of mites, selection of patients with mite allergy by clinical history is difficult. METHODS: A total of 210 Cuban asthmatics with continuous symptoms, and 43 Swedes with a clinical history of mite allergy were investigated. Skin prick tests were performed with D. siboney, D. pteronyssinus, D. farinae, B. tropicalis, Acarus siro, Lepidoglyphus destructor, and Tyrophagus putrescentiae extracts. For analysis of the biologic activity of mite extracts, Cuban patients were divided into four groups: 1) all patients skin-test-positive to mites 2) patients positive to mites, but not to other inhalant allergens 3) patients reacting most to the mite species analyzed 4) patients reactive only to mites and reacting most to the mite species analyzed. The biologic potency was calculated according to the Nordic Guidelines. RESULTS: Due to cross-reactivity between mites, Swedish mite sensitive patients, with a clear clinical history of mite allergy, but not exposed to D. siboney and B. tropicalis, were more skin reactive to these mites than were Cubans. The estimated potency increased gradually to >200% in group 4. In group 1 Cubans, the reactivity to all mites but B. tropicalis was lower than that in mite-sensitive Swedes. CONCLUSIONS: According to the influence of patient selection criteria on the estimation of the potency of mite extracts, the determination of the biologic activity of allergenic extracts in subjects without a clear-cut clinical history should be replaced by new methods when available. PMID- 10371101 TI - Pseudoallergy or nonallergic hypersensitivity. PMID- 10371102 TI - Multiple vaccination effects on atopy. PMID- 10371103 TI - Evolution from ewe's milk to cow's milk allergy. PMID- 10371104 TI - Asthma and TNF variants in Chinese and Malays. PMID- 10371105 TI - Vacuum cleaners and airborne dog allergen. PMID- 10371106 TI - Occupational rhinitis caused by beech wood dust. PMID- 10371107 TI - Renal failure--a new risk group for latex allergy? PMID- 10371108 TI - Mite allergens in feather and synthetic pillows. PMID- 10371109 TI - Early angiographic results after revascularization by minimally invasive direct coronary artery bypass (MIDCAB). AB - OBJECTIVE: Anastomosing the left anterior descending artery (LAD) by use of the internal mammary artery (IMA) via an anterior minithoracotomy represents the most commonly performed minimally invasive direct coronary artery bypass (MIDCAB). However, little is known about angiographic results beyond patency rates. METHODS: Therefore, a retrospective analysis of 205 consecutive control angiographies was performed evaluating anastomototic patency as well as the angiographic morphology of the left IMA and the LAD. RESULTS: The overall anastomotic patency rate was 98.0%. As a result of incomplete IMA preparation (6 15 cm) large side branches (n = 4), or an IMA course under tension (n = 6) were occasionally observed. Despite a tension-free course, the IMA appeared fixed to the chest wall without functional compromise in 21 cases. A restrictive thrombus formation occurred once, IMA dissection was not seen. Two of the grafts developed spasms. A distal IMA stenosis >50% was seen in five cases. Stenosis of the anastomosis (>50%) itself was found once, as well as unexpected malinsertation to diagonal branches (n = 4). Compared with preoperative angiograms, de novo stenoses of the LAD were assessed proximal (14< or =50%, 2>50%) and distal (15< or =50%, 2>50%) to the anastomosis. Elevation of the LAD out of the vascular bed was an additional finding (n = 12) in a few cases. CONCLUSIONS: The overall patency of MIDCAB-LAD-grafting appears to be equivalent to conventional IMA grafting to the LAD. Particular angiographic findings, however, may be directly associated to the applied surgical technique. PMID- 10371110 TI - Detection of coronary artery bypass graft patency by contrast enhanced magnetic resonance angiography. AB - OBJECTIVE: The subject and purpose of the prospective study was to delineate coronary artery bypass graft (CABG) course and to determine patency of aortocoronary venous bypass grafts (ACVB) compared with internal mammary artery bypass grafts (IMA) in the early postoperative follow-up, by contrast enhanced magnetic resonance angiography (MRA). For control, patients were examined with X ray angiography and spiral-computed tomography (CT). METHODS: Eighty-five patients (74 male/11 female) with a mean age of 63.7 years underwent MRA examination, applying contrast enhanced gradient-echo sequence after an average distance of 7 days from CABG surgery. A 1.5 Tesla magnetom vision (Siemens, Erlangen, Germany) with phased array coil technology was used. Overall, 247 bypass grafts (160 ACVB/87 IMA) were studied with a 3D (three dimensional) ultrashort TE gradient-echo sequence (TR/TE/a:5 ms/2 ms/40 degrees) with 512*512 matrix and 500 mm FoV in single breath-hold technique after Gd-DTPA bolus injection. CABGs were judged in three different parts, including the course of CABG and both anastomoses. CABGs were controlled by angiography and spiral-CT to examine sensitivity, specificity and efficiency of MRA examination. Additional measurement of bypass graft flow velocity of arterial and venous grafts was performed with 2D phase contrast technique in breath-hold technique with ECG triggering. RESULTS: One hundred and thirty-nine of 160 (86.9%) ACVB grafts and 83 of 87 (95.4%) IMA grafts could be visualized. Suspected occlusions of 10 CABGs were confirmed in 80% with a second modality. Five CABGs were false positive in MRA. MRA proved a high specificity (93.8%), sensitivity (89.9%) and efficiency (1.73), especially in detection of IMA to LAD and ACVB to LAD and RCA (Table 1). 3D maximum intensity projection (MIP) reconstruction was helpful in delineating CABG course and in several cases in detecting stenosis of coronary arteries. Results of flow velocity showed a significant higher mean systolic velocity in arterial bypasses than in venous grafts with a higher maximum velocity in systole than in diastole in both grafts. Bypass stenosis in distal anastomosis could not be verified with MRA and flow method. CONCLUSION: Contrast enhanced 3D ultrashort TE gradient-echo magnetic resonance angiography has the potential for being a reliable method for CABG visualization and CABG patency determination in the early postoperative period. MR flow measurement was not qualified for detection of a bypass stenosis. PMID- 10371111 TI - Preoperative lipid-control with simvastatin reduces the risk of postoperative thrombocytosis and thrombotic complications following CABG. AB - OBJECTIVE: It has earlier been suggested that postoperative thrombocytosis frequently occur after coronary artery bypass grafting (CABG) and may be linked to lipid disturbances. A prospective randomized study was undertaken to evaluate if preoperative lipid-control, using HMG-CoA-reductase inhibitor (Zocor), simvastatin, reduces the risk of postoperative thrombocytosis. METHODS: Seventy seven patients with symptomatic coronary artery disease and hypercholesterolemia (total cholesterol > or =6.2 mmol/l), planned for CABG where randomly assigned to; undergo CABG without preoperation lipid control (group I, n = 37) or undergo simvastatin-treatment (20 mg daily) to control their lipids (4 weeks) prior to CABG (group II, n = 40). RESULTS: Patient characteristics and operation data did not differ between the groups. Serum-cholesterol, cholesterol/HDL-cholesterol, LDL-cholesterol, Apolipoprotein A1 and Plasminogen were all significantly higher in group I patients compared with group II just prior to surgery. Other laboratory parameters did not differ. RESULTS: In group II, total cholesterol and cholesterol/HDL-cholesterol quota were significantly lowered by simvaststin (-2 and -29%, respectively). Postoperative thrombocytosis (platelet counts > or =400000/microl) occurred significantly more frequently in group I 81% (30/37) compared with 3% (1/40) in group II, P<0.0001. Myocardial infarction after the 7th postoperative day was more often diagnosed in group I, 14 vs. 0% in group II. Postoperative transient renal failure occurred also more frequently in group I, 24% compared with 8% in group II. Other postoperative complications and laboratory data did not differ. CONCLUSIONS: This study once again underlines the importance of lipid control using HMG-CoA-reductase inhibitors (e.g. Zocor) in patients with established coronary artery disease. For the first time it is shown that lipid-control with simvastatin prior to CABG reduces the risk of postoperative thrombocytosis, thus lowers the risk for thrombotic complications. PMID- 10371112 TI - The meaning of early mortality after CABG. AB - OBJECTIVE: Investigations of early mortality after coronary artery bypass grafting (CABG) are predominantly based on 30-day mortality or hospital mortality. The advantages, disadvantages, and usefulness of hospital mortality and 30-day mortality analyses to investigate the early risk after CABG are evaluated. METHODS: A total of 4985 patients underwent isolated CABG from June 1988 to June 1997. A follow-up was performed 180 days after CABG (response rate: 98.6%). RESULTS: The mean hospital stay was 13.5+/-9.6 days, the range was 0 to 142 days (25% quartile, 9 days; median, 12 days; 75% quartile, 15 days). The hospital mortality was 5.3%. The 30-day mortality was 5.6%. The non-parametric Kaplan-Meier curve of the time interval 0-180 days postoperatively proves the persistence of the still decreasing behaviour of the survival curve beyond the 30th day until about the 60th postoperative day. Stratified by era of operation, the 'early phase' after CABG seems to be prolonged beyond 30 days at least for the more recent operation era since 1991. Risk stratification proves that the higher the risk group, the more the early phase tends towards a prolongation. CONCLUSIONS: The hospital mortality reflects institutional habits concerning postoperative patient care. Therefore, a systematic underestimation of early mortality is likely. In contrast to hospital stay, the evaluation of 30-day mortality requires a follow-up procedure but allows interinstitutional comparisons. Nevertheless, 30-day mortality systematically underestimates the early risk, at least in the more recent CABG period. So, a standardized evaluation of a longer time period (p.e. 180 days) is recommended. PMID- 10371113 TI - Endothelin-1 is involved in plasma mediated stimulation of neutrophil adherence during coronary artery bypass grafting. AB - OBJECTIVE: Myocardial ischaemia followed by reperfusion during coronary artery bypass grafting (CABG) is known to result in the activation of polymorphonuclear neutrophils (PMN). The activation of PMN during ischaemia/reperfusion may be a result of their direct contact with activated endothelial cells and/or an effect of stimuli released from ischaemic myocardium. Increased expression of adhesion molecules on the PMN surface, after activation, leads to coronary capillary plugging with a subsequent decrease in blood flow. The purpose of the study was to evaluate plasma-mediated stimulation of PMN adhesion during CABG and to verify if endothelin-1 (ET-1), known to be a potent stimulus for PMN, is involved in stimulation of neutrophils adhesion mediated by integrins. METHODS: Coronary sinus, peripheral artery and peripheral venous plasma samples were taken from 11 patients undergoing coronary surgery before aortal cross-clamping, at the beginning of reperfusion and 30 min thereafter. PMN isolated from five healthy volunteers were incubated with the plasma (20 samples per patient) in the presence of saline or a specific ET-1 receptor blocker, and PMN adherence to a microtiter plate covered with a monoclonal antibody against CD 18 antigen (beta subunit of the integrin family of adhesion molecules) was evaluated. RESULTS: We have observed a significant increase in adhesion of PMN incubated in the presence of saline with the plasma taken from coronary sinus at the beginning of reperfusion (7.79+/-1.64% of adhering cells) as compared with plasma obtained before aortal cross-clamping from the same place (6.78+/-1.3%, P = 0.04) and from peripheral artery at the beginning of reperfusion (6.64+/-1.1%, P = 0.04, means +/- SEM). ET-1 receptor blocker, significantly decreased stimulation of PMN adhesion by coronary sinus plasma obtained at the beginning of reperfusion (6.7+/ 1.51%, P = 0.02). Plasma levels of ET-1 (ELISA) in the samples taken from coronary sinus at the beginning of reperfusion, were higher than in samples obtained before myocardial ischaemia or 30 min after reperfusion. CONCLUSIONS: We conclude, that soluble stimuli capable of stimulation of PMN adhesion are released following myocardial ischaemia during CABG and ET-1 may be involved in PMN stimulation. PMID- 10371114 TI - Surgical treatment of left ventricular post-infarction aneurysm with endoventriculoplasty: late clinical and functional results. AB - OBJECTIVE: The temporal response to endoventriculoplasty (EVP) has not been well defined. We have evaluated the long-term clinical and functional results of this technique. METHODS: From 1988 to 1997, 121 patients underwent aneurysmectomy by EVP associated with myocardial revascularization for anteroapical left ventricular postinfarction aneurysm. Among these, 39 patients (43%) underwent early post-operative cardiac catheterization (within 3 months maximum), and were available to be revaluated after a mean follow-up time of 56+/-28 months, by means of a new hemodynamic study. Left ventricular silhouettes were analyzed by means of a special software. RESULTS: The mean New York Heart Association functional class decreased from 2.5+/-0.9 to 1.6+/-0.8 (P<0.001) late postoperatively. The global ejection fraction improved early postoperatively from 43+/-13 to 61+/-13% (P<0.001), and late postoperatively slightly decreased to 42+/-13% (ns) versus preoperative values. Left ventricular end diastolic pressure early postoperatively fell from 16.8+/-7 to 15.7+/-6.7 (ns), and late postoperatively increased to 21.6+/-8.8 (ns) versus preoperative values. Pulmonary artery pressure rose early postoperatively from 31.5+/-6.4 to 32.1+/ 6.7 (ns), and late postoperatively to 34.9+/-8.9 (ns). The global contractility score decreased early postoperatively from 42.3+/-9.6 to 28.4+/-13.6 (P<0.001); the global late postoperative contractily was 35+/-14 (ns) versus preoperative values. Patients who benefit most from the operation were those with a normal postoperative contraction pattern, where ejection fraction improved respectively early postoperatively from 43+/-13 to 63+/-11% (P<0.001), and late postoperatively to 49+/-10% (P<0.001) versus preoperative values. Occlusion or critical stenosis of bypass grafts occurred in 10 patients (25.6%). There were no significant differences in hemodynamic data and hypokinesis score changes between patients with patent or occluded bypass graft, and between patients with mono or multivessel disease. The operative mortality was 6.3%, and 8.8% needed intraaortic balloon counterpulsation. The actuarial survival rates at 5 and 7 years were 73+/-6 and 61+/-6%. The mean follow-up period was 68 months (with 112 months maximum). CONCLUSIONS: We conclude that, in our patients group, EVP of left ventricular aneurysm associated with coronary grafting improves clinical status after operation. We registered a trend for a mild hemodynamic worsening, irrespective of coronary artery disease except in those patients who had shown a normal postoperative contraction pattern. PMID- 10371115 TI - The hemodynamic effects of double-orifice valve repair for mitral regurgitation: a 3D computational model. AB - OBJECTIVES: A 3D computational model has been implemented for the evaluation of the hemodynamics of the double orifice repair. Critical issues for surgical decision making and echo-Doppler evaluation of the results of the procedure are investigated. METHODS: A parametric 3D computational model of the double-orifice mitral valve based on the finite elements model has been constructed from clinical data. Nine different geometries were investigated, corresponding to three total inflow areas (1.5, 2.25 and 3 cm2) and to three orifice configurations (two equal orifices, two orifices of different areas, i.e. one twice as much the other one, and a single orifice). The simulations were performed in transit; the fluid was initially quiescent and was accelerated to the maximum flow rate with a cubic function. For each case, some characteristic values of velocity and pressure were determined: velocities were calculated downstream of each orifice, at the centre of it (Vcen1, Vcen2). The maximum velocity was also determined for each orifice (Vmax1, Vmax2). Maximum pressure drops (deltap(max)) across the valve were compared with the estimations (deltap(Bernoulli)) based on the Bernoulli formula (4 V2). RESULTS: In each simulation, no notable difference was observed between Vcen1 and Vcen2, and between Vmax1 and Vmax2, regardless of the valve configuration. Maximum velocity and deltap(max) were related to the total orifice area and were not influenced by the orifice configuration. Deltap(Bernoulli) calculated with Vmax was well correlated with the deltap(max) obtained throughout the simulations (y = 0.9126x + 0.3464, r = 0.996); on the contrary the pressure drops estimated using Vcen underestimated (y = 0.6757x + 0.3073, r = 0.999) the actual pressure drops. CONCLUSIONS: The hemodynamic behaviour of a double orifice mitral valve does not differ from that of a physiological valve of same total area: pressure drops and flow velocity across the valve are not influenced by the configuration of the valve. Echo Doppler estimation of the maximum velocities is a reliable method for the calculation of pressure gradients across the repaired valve. PMID- 10371116 TI - Survival and prognostic factors in patients undergoing parenchymal saving bronchoplastic operation for primary lung cancer: a series of 110 consecutive cases. AB - OBJECTIVE: The purpose of this study was to report our experience concerning bronchial sleeve lobectomy for treating bronchogenic cancer. METHOD: From 1980 to 1994, 110 patients underwent bronchial sleeve lobectomy for bronchogenic cancer. In 45 patients, preoperative investigations contraindicated pneumonectomy, whereas in 65 other patients, sleeve resection was performed without functional necessity. The most common procedures were sleeve lobectomy of the right upper lobe (64%), and of the left upper lobe (21%). Sixteen patients (15%) underwent additional arterial vascular resection. Seven patients had microscopic invasion of the bronchial margin without the possibility of further resection in six with regard to their limited respiratory function. Tumors were staged as follow: 32 stage IB (all T2 N0), 57 stage IIB (57T2 N1), and 17 stage IIIA (eight, T3N1; nine, T2N2), whereas four patients had an in situ cancer (four stage 0). RESULTS: Operative mortality was 2.75%. The 5- and 10-year actuarial survival rates were, respectively, 39 and 22% for the entire group. The 5-year actuarial survival rates were, 60% in stage IB, 30% in stage IIB, and 27% in stage IIIA. Four factors significantly influenced survival (P<0.05): nodal stage, arterial resection, invasion of the bronchial stump and poor functional respiratory status contraindicating pneumonectomy. CONCLUSIONS: In our experience, sleeve resection for stage I provides comparable survival to that of standard resection at equal stage. However, in patients with pathologically N1 disease, who can tolerate a pneumonectomy, a randomized study is mandatory to confirm that sleeve lobectomy can be performed without the risk of decreasing long-term survival. In our study, patients who required an associated vascular resection demonstrated a poor survival. PMID- 10371117 TI - Time trend in the surgical management of patients with lung carcinoma. AB - OBJECTIVE: The goal of the study was to analyze the histological and clinical trends in lung carcinoma and their influence upon the preoperative evaluation, surgical procedures and survival. METHODS: We retrospectively reviewed the charts of 1079 consecutive patients who underwent surgery for primary lung carcinoma between 1977 and 1996 in our institution. Patients were divided into five equal 4 year periods according to the year of surgery (1977-1980; 1981-1984; 1985-1988; 1989-1992; 1993-1996). RESULTS: Between 1977-1980 and 1993-1996, the incidence of squamous cell carcinoma significantly declined, whereas the incidence of adenocarcinoma and bronchioloalveolar carcinoma increased. During the same period, the proportion of squamous cell carcinoma visualized at bronchoscopy and the rate of preoperative histological diagnosis significantly decreased. An increasing proportion of lobectomy and less extended resection was associated with an increasing number of patients with stage I carcinoma. Meanwhile, the operative mortality significantly declined from 9 to 4% and the 5-year survival improved from 25 up to 40%. CONCLUSION: Over the last two decades, the shift in histological distribution was associated with an increasing proportion of patients with stage I disease, a lower operative mortality and a better 5-year survival. PMID- 10371118 TI - Postoperative adjuvant chemotherapy with PVM (Cisplatin + Vindesine + Mitomycin C) and UFT (Uracil + Tegaful) in resected stage I-II NSCLC (non-small cell lung cancer): a randomized clinical trial. West Japan Study Group for lung cancer surgery (WJSG). AB - OBJECTIVE: The West Japan Study Group For Lung Cancer Surgery (WJSG) conducted a randomized controlled trial in order to assess the usefulness of adjuvant chemotherapy for NSCLC. METHODS: Patients with completely resected NSCLC (stages I and II) were enrolled in the trial. These patients were randomized into two groups: a surgery alone group; and a chemotherapy group which received intravenous administration of two courses of 4-week PVM chemotherapy (80 mg/m2 of Cisplatin on day 1, 2-3 mg/m2 of Vindesine on day 1 and/or day 8, and 8 mg/m2 of Mitomycin C on day 1), after which they took 400 mg/day of UFT (Uracil + Tegaful) orally for 1 year. RESULTS: Among 229 patients registered for the study from August 1988 to July 1990, 225 were available cases (116 patients in the surgery alone group, and 109 patients in the chemotherapy group). No bias in prognostic factors could be found between the two groups. The 5-year survival rate was 71.1% for the surgery-alone group and 76.8% for the chemotherapy group with no significant difference observed. However, subset analysis demonstrated that PVM therapy improved the post operative survival of pT1N0 patients (the 5-year survival rate: 75.3% for the surgery alone group, and 90.7% for the chemotherapy group P<0.05). CONCLUSIONS: It is interesting to find that among pT1N0 patients, who were not regarded as a target of chemotherapy, those receiving chemotherapy showed significantly better prognostic results. These findings suggest the necessity of further studies on the adjuvant chemotherapy, even in the early stages. PMID- 10371119 TI - Right ventricular morphology and function after pulmonary resection. AB - OBJECTIVE: To identify the effect of pulmonary resection on right ventricular performance and its possible contribution to mortality and morbidity. METHODS: Before and 2 days after pulmonary resection for primary lung cancer in 31 patients (21 males; ages 32-69 years), echocardiographic examinations of the right ventricle were performed. Systolic, diastolic and stroke volumes as well as right ventricular ejection fraction were estimated. Right ventricular volumes were calculated using the subtracting method. RESULTS: Right ventricular end diastolic volume index increased significantly in patients after pneumonectomy: 80.4+/-7.2 ml/m2 versus preoperative evaluation: 66.1+/-5.2 ml/m2 (P = 0.031). In patients who underwent pneumonectomy right ventricular ejection fraction significantly decreased from 48+/-5.0% preoperatively to 39%+/-4.1% after surgery (P = 0.027). Fourteen patients after pneumonectomy had development of supraventricular arrhythmias postoperatively. These patients had much higher right ventricular end-diastolic volume index (76.3+/-6.4/82.1+/-7.4; P = 0.032) and lower right ventricular ejection fraction (42+/-4.3/37+/-3.9; P = 0.021) after surgery in comparison with patients who had normal sinus rhythm postoperatively. CONCLUSION: Pulmonary resection caused a significant dilatation and dysfunction of right ventricle in the early postoperative period. Early detection of deterioration in right ventricular function after pneumonectomy may provide the opportunity for interventional therapy. PMID- 10371120 TI - Ventilatory muscle recruitment and work of breathing in patients with respiratory failure after thoracic surgery. AB - OBJECTIVES: Increased work of breathing (WOB) and respiratory muscle weakness have been identified as major causes of respiratory failure after thoracic surgery. This study was undertaken firstly to characterize the mechanical impairment in patients with respiratory failure after cardio-thoracic surgery, and secondly, to determine how diaphragmatic paralysis affects deterioration in the ventilatory mechanics. METHODS: We evaluated the respiratory mechanics of 24 patients following cardiac and thoracic surgery. Ten patients without respiratory problems were examined as control subjects. There were nine patients with phrenic nerve injury and five patients without phrenic nerve injury who required mechanical ventilation for more than 7 days. Phrenic nerve injury was assessed with a phrenic nerve stimulation test. We measured the respiratory variables, the esophageal, gastric and transdiaphragmatic pressure swing (deltaPes, deltaPga and deltaPdi, respectively), and the work of breathing during quiet tidal breathing. RESULTS: Both the groups requiring mechanical ventilation exhibited abnormally negative deltaPga/deltaPes values, compared with the control subjects. A significant increase in WOB with the normal generation of deltaPdi was seen in the patients without phrenic nerve injury. In contrast, the poor generation of deltaPdi with a slight increase in work of breathing was noted in patients with phrenic nerve injury. CONCLUSIONS: These results demonstrated two different types of respiratory failure in thoracic surgery patients, focusing on the impact of phrenic nerve paralysis. Diaphragmatic dysfunction should not be overlooked in postoperative care, and the amelioration of this compromise in respiratory mechanics is an important aspect of good patient management. PMID- 10371121 TI - Surgical treatment of primary pulmonary sarcomas. AB - OBJECTIVE: We sought to identify the long-term prognosis after surgical treatment for primary pulmonary sarcoma. METHODS: Twenty-three patients were retrospectively identified as having been treated surgically for primary pulmonary sarcoma between 1981 and 1996. The records of all patients were reviewed, and the histopathology reexamined by a pathologist. RESULTS: Fifteen patients were male and eight female; their ages ranged from 20 to 78 (mean 51) years. Tumors measured between 0.9 and 12.0 (mean 5.2) cm across the greatest diameter. The histologic diagnoses were malignant fibrous histiocytoma (8, three grade 1 or 2, two grade 3), synovial sarcoma (4), malignant schwannoma (3), leiomyosarcoma (3), and one case each of angiosarcoma, intimal sarcoma, epitheloid hemangioendothelioma, fibrosarcoma and primitive neuroectodermal tumor. Three patients were found to be unresectable. All three underwent radiation and chemotherapy. Lobectomies or bilobectomies were performed in 13 patients including two sleeve resections, one carinal resection, and one chest wall resection. Four patients underwent radical pneumonectomies. Three patients with invasion of the pulmonary artery, pulmonary veins or atrial wall underwent extended resections with the use of cardiopulmonary bypass. In two, a homograft was used to reconstruct the right ventricular outflow tract. Of the resected patients, six had a positive resection margin, and four had at least one positive lymph node in the specimen. Three patients underwent repeat pulmonary resections for recurrences. Eleven patients received postoperative chemotherapy and eight had radiation therapy. Follow-up was available on 22 patients, and ranged from 2 to 183 (mean 48) months; 14 patients are disease free, six died of disease, one died of surgical complications (operative mortality 5%), and two are alive with disease. Actuarial 3- and 5-year survival of the resected patients was 69%. Size and grade were not found to be correlated with significantly increased survival, but completeness of resection was (P<0.05). CONCLUSIONS: Resection of primary pulmonary sarcomas can produce an acceptable survival rate if the resection is complete. Cardiopulmonary bypass can be a useful adjunct when tumors involve a resectable area of the heart or great vessels. PMID- 10371122 TI - Glove perforation rate in open lung surgery. AB - OBJECTIVE: In open lung surgery the surgical access is encircled by the ribs, which should result in a high glove perforation rate compared with other surgical specialities. METHODS: Prospectively the surgeon, first and second assistant and the scrub nurse wore double standard latex gloves during 100 thoracotomies. Parameters recorded were: procedure performed, number of perforations, localization of perforation, the seniority of the surgeon, manoeuvre performed at the moment of perforation, immediate cause of perforation, operation time, performance of rib resection during thoracotomy and time of occurrence of the first three perforations. RESULTS: One thousand, six hundred and seventy-three gloves (902 outer, 771 inner) were tested. In 78 operations perforations occurred. There were 150 outer glove perforations (8.9%, 0-8, mean 1.23), 19 inner glove perforations (1.13%, 0-2, mean 0.19). Cutaneous blood exposure was prevented in 78% of all operations and in 87% of all perforations. The perforation rate for the surgeon, the scrub nurse, the first and the second assistant were 61.2, 40.4, 9.7 and 3.1% of all operations, respectively. Rib resection and a duration of more than 2 h resulted in a significant rise of glove perforation rate (P<0.05). The personal experience of the surgeon and the type of operation did not correlate with glove perforation. The immediate cause leading to perforation was named in only 17 cases (13.7%) and comprised contact with bone (seven), a needle stitch (seven) and a production flaw (three). Leaks were localized mostly on the first finger (18%),second finger, (39%) palm and dorsum of the hand (16%). The average occurrence of all first perforations was 38.7 min (range 3-190) after the beginning of surgery, the second after 63.2 min (range 10 195). Fifty-four first perforations (50.5%) were found during the first 30 min of the operation. CONCLUSIONS: The reported perforation rate of 78% lies in the highest range of reported perforation rates in different surgical specialities. Double gloving effectively prevented cutaneous blood exposure and thus should become a routine for the thoracic surgeon to prevent transmission of infectious diseases from the patient to the surgeon. PMID- 10371123 TI - The possibilities of greater omentum usage in thoracic surgery. AB - OBJECTIVE: To illustrate the wider role of an omental pedicle flap in surgical treatment for diverse thoracic organ disorders we have, retrospectively, reviewed our experience over the last 8 years. METHODS: We used the greater omental pedicle flap in 68 patients. Bronchial stump omentopexy was performed in 35 patients with a high risk of impaired bronchial healing after right pneumonectomy as a preventive method and in two patients with an acute bronchial fistula. In 13 patients after circular tracheal or carinal resections and in four after esophago respiratory disjoints of fistulas we applied anastomoses or circular omentopexy of the fistula zone. The omental coverage was performed in four patients with chronic empyema, in seven patients after extensive chest wall resection and in two patients with post-sternotomy mediastinites. In one patient with idiopathic fibrosing mediastinitis, one-stage allotransplantation of tracheal thoracic segment was conducted and greater omentum (GO) was used to wrap the allograft. RESULTS: Three patients developed major complications. The first, bronchial fistula after bronchial stump omentoplasty. In two patients circular wide tracheal resection (11-12 rings) was complicated by a tracheal divergence of anastomoses. These complications were cured in a conservative way since a displaced omental flap substituted the tracheal wall and, therefore, the trachea remained hermetically sealed. In other cases, the perfect adhesive properties of the omental tissue promoted perfect adhesive properties promoted prevention of incompetence and inflammatory complications. Immunological and bacteriological examinations showed that pediculated omental flap is a lymphocyte source and promotes a decrease in bacterial quantity and activity. CONCLUSION: We think the series demonstrates the value of the omental method, which offers excellent therapeutic results following an easy surgical procedure. This method extends indications for surgical treatment and decreases the postoperative complications. PMID- 10371124 TI - Blunt diaphragmatic rupture. AB - OBJECTIVE: To identify (1) predictors of outcome in blunt diaphragmatic rupture (BDR), and (2) factors contributing to diagnostic delay. METHODS: We reviewed the charts and radiographs of 41 patients with BDR treated in our Hospital from 1988 to 1997. There were 35 male (85%) and six female, aged 17-71 (mean: 41) years. BDR was left-sided in 24 cases (58%), right-sided in 15 (36%) and bilateral in two (5%). RESULTS: Two groups of patients can be identified: group A (n = 36, 88%) with acute BDR, and group B (n = 5, 12%) with post-traumatic diaphragmatic hernia (TDH). In group A, immediate diagnosis was made in 35 cases (97%), but only in 26 (72%) preoperatively. In one case, a right BDR was missed on initial evaluation but became apparent 2 weeks later. Associated injuries were present in 34 patients (94%) involving: spleen (n = 18), rib fractures (n = 17), liver (n = 14), lung (n = 11), bowel (n = 7), kidney (n = 5) and other fractures (n = 21). Injury Severity Score (ISS) ranged from 9 to 66 (mean: 31). BDR repair was accomplished through a laparotomy in 22 cases, thoracotomy in 10 and laparo thoracotomy in four. The overall mortality rate was 16.6% (6/36). Both patients with bilateral BDR died. The patients who died were older than the survivors (mean age: 54 vs. 39 years, P<0.05), were more severely injured (mean ISS: 46 vs. 28, P<0.05) and were in shock (100 vs. 23%, P<0.05). In group B with TDH, diagnosis was delayed for 7-16 months after injury. Four patients had non specific clinical signs and one strangulation of hollow viscera. One patient had undergone surgery during acute injury but BDR was overlooked. Location of TDH was on the left in three cases and on the right in two. Delay in BDR diagnosis was 12.5% (3/24) in patients with left-sided and 20% (3/15) in patients with right sided lesions (P>0.1). Repair of TDH was achieved through thoracotomy in all cases. No mortality or major morbidity were encountered. CONCLUSIONS: (1) Predictors of BDR mortality are: age, ISS and hemodynamic status of the patient. (2) Delay in diagnosis does not influence the outcome and is not influenced by the side of BDR location. (3) BDR can easily be missed in the absence of other indications for prompt surgery, where a thorough examination of both hemidiaphragms is mandatory. A high index of suspicion combined with repeated and selective radiologic evaluation is necessary for early diagnosis. PMID- 10371125 TI - Donor and recipient treatment with the Lazaroid U-74006F do not improve post transplant lung function in swine. AB - OBJECTIVE: U-74006F is the only Lazaroid which is currently in clinical use. A number of experimental studies demonstrate that Lazaroids reduce ischemia/reperfusion injury in various organ systems. We evaluated the effect of U-74006F on reperfusion injury in a large animal model of lung allo transplantation. METHODS: Two different treatment modalities were evaluated and compared with corresponding control groups. Unilateral left lung transplantation was performed in 21 weight-matched pigs (24-31 kg). Donor lungs were flushed with 1.51 cold (1 degrees C) LPD solution and preserved for 20 h. In group I (n = 5), donor animals were pretreated with U-74006F (10 mg/ kg i.v.) 20 min before harvest. In addition U-74006F was added to the flush solution (10 mg/l). In group III (n = 6), the Lazaroid was given to the donor before flush and to the recipient before reperfusion (3 mg/kg i.v.). Group II and IV (n = 5) served as control. One hour after reperfusion, the recipient contralateral right pulmonary artery and bronchus were ligated to assess graft function only. Extravascular lung water index (EVLWI), mean pulmonary artery pressure, cardiac output, and gas exchange were assessed during a 5 h observation period. Lipid peroxidation (TBARS) and neutrophil migration (MPO activity) were measured at the end of the assessment in lung allograft tissue. RESULTS: A significant change of TBARS concentration was shown in group III (group III 78.7+/-4.6 pmol/g vs. group IV 120.8+/-7.2 pmol/g (P = 0.0065) normal lung tissue 41.3+/-4.2 pmol/g). MPO activity was reduced in group III 3.74+/-0.25 deltaOD/mg per min vs. group IV 4.97+/-0.26 deltaOD/mg per min (P = 0.027), normal lung tissue 1.04+/-0.27 deltaOD/mg per min). Pulmonary hemodynamics and gas exchange after reperfusion did not differ between groups. In group I and III, a tendency towards a reduced EVLWI was noted. CONCLUSION: We conclude that combined treatment of donor and recipient with U-74006F reduces free radical mediated injury in the allograft. However, this intervention did not result in a significant reduction of post transplant lung edema or improvement of pulmonary hemodynamics. Donor pretreatment alone did not improve lung allograft reperfusion injury. These results indicate that the benefit of U-74006F is too small to consider clinical application in lung transplantation. PMID- 10371126 TI - Influence of different routes of flush perfusion on the distribution of lung preservation solutions in parenchyma and airways. AB - OBJECTIVE: The present study was performed to investigate the influence of different routes of perfusion on the distribution of the preservation solutions in the lung parenchyma and upper airways. METHODS: Pigs were divided into four groups: control (n = 6), pulmonary artery (PA) (n = 6), simultaneous PA + bronchial artery (BA) (n = 8), and retrograde delivery (n = 6). After preparation and cannulation, cardioplegia solution and Euro-Collins solution (ECS) for lung preservation were given simultaneously. After removal of the heart, the double lung bloc was harvested. Following parameters were assessed: total and regional perfusion (dye-labeled microspheres), tissue water content, PA, aorta, left atrial and left ventricular pressures, cardiac output and lung temperature. RESULTS: Our data show that flow of the ECS in lung parenchyma did not reach control values (9.4+/-1.0 ml/min per g lung wet weight) regardless of the route of delivery (PA 6.3+/-1.5, PA + BA 4.8+/-0.9, retrograde 2.7+/-0.9 ml/min per g lung wet weight). However, flow in the proximal and distal trachea were significantly increased by PA + BA delivery (0.970+/-0.4, respectively, 0.380+/ 0.2 ml/min per g) in comparison with PA (0.023+/-0.007, respectively, 0.024+/ 0.070 ml/min per g), retrograde (0.009+/-0.003, respectively, 0.021+/-0.006 ml/min per g) and control experiments (0.125+/-0.0018, respectively, 0.105+/ 0.012 ml/g per min). Similarly the highest flow rates in the right main bronchus were achieved by PA + BA delivery (1.04+/-0.4 ml/min per g) in comparison with 0.11+/-0.03 in control, 0.033+/-0.008 in PA, and 0.019+/-0.005 ml/min per g in retrograde group. Flows in the left main bronchus were 0.09+/-0.02 ml/min per g in control, 0.045+/-0.012 ml/min per g in PA, and 0.027+/-0.006 ml/min per g in retrograde group. The flow rates were significantly (P = 0.001) increased by PA + BA delivery of the storage solution (0.97+/-0.3 ml/min per g). CONCLUSIONS: Our data show that the distribution of ECS for lung preservation is significantly improved in airway tissues (trachea and bronchi) if a simultaneous PA + BA delivery is used. PMID- 10371127 TI - Lung infections in pediatric lung transplantation: experience in 49 cases. AB - OBJECTIVES: Pulmonary infections, and particularly cytomegalovirus (CMV) infections, are a major cause of morbidity after lung transplantation. We report here our results in 49 pediatric lung transplantations. METHODS: Between may 1988 and 1997, we have done 49 lung transplantations in 42 children (en bloc double lung transplantation (DLT):10, HLTx:7, sequential bilateral sequential-lung transplantation (BSLT):31, single-lung transplantation (SLT): 1). In seven, it was a retransplantation. Among these, 34 were cystic fibrosis (CF) patients, all with multiresistant organisms (Pseudomonas aeruginosa, Burkholderia cepacia, Achromobacter xylososydans, Staphylococcus aureus). All patients were treated with multiantibiotic prophylaxy adapted to the preoperative cultures. Donor recipient CMV matching was possible in only 31 cases. CMV prophylaxy and immunosuppression protocols have evolved with time, with a current protocol of IV Gancyclovir prophylaxy for 3 months and triple drug immunosuppression without post-operative rabbit anti-thymocyte globulin (RATG) induction. There was no perioperative mortality in the primary transplantations and three early deaths in the whole group (6.1%). RESULTS: Only five patients had no pulmonary infection. The patients presented 3.2 infection episodes per year, 75% localized on the lungs, 41% during the first 3 months. Among the 13 deaths in the 1st year, 10 were directly related to infection, 60% due to CMV. After the 1st year, in all patients dying of pulmonary dysfunction or obliterative bronchiolitis (OB), bacterial infections were associated. There was no serious fungal infection. Actuarial survival at 3 months, 1, 3, 5 years were 85, 65.7, 47.5 and 28.5%, respectively. There was a significant difference in 3 year survival between patients receiving CMV negative organs (40%) and CMV positive organs (17%). CONCLUSION: In our experience, as in other's, pulmonary infection risk is important in lung transplantation. Bacterial infections were mainly an aggravating factor of secondary pulmonary dysfunction or OB, and were not the primary cause of death. CMV infections have been very severe and lead us, despite the scarcity of donors, to avoid positive donors in negative recipients, this leads to disastrous mid-term results in our experience, despite prophylaxis. PMID- 10371128 TI - Different techniques of distal aortic repair in acute type A dissection: impact on late aortic morphology and reoperation. AB - OBJECTIVE: To compare three different techniques of distal aortic repair in acute type A (de Bakey type I) aortic dissection and to evaluate their impact on the late morphology of the aortic arch and descending aorta and on the incidence of reoperation. METHODS: From 65 patients operated on due to an acute type A aortic dissection between 1989 and 1993, 54 long-term survivors underwent clinical and radiologic follow-up examination after a mean postoperative interval of 62+/-16 months. The surgical techniques of distal aortic reconstruction included closed repair using Teflon felt reinforcement under moderate hypothermic cardiopulmonary bypass (n = 20) and open repair in deep hypothermic circulatory arrest using either Teflon felt reinforcement (n = 16) or gelatin-resorcin-formaldehyde (GRF) glue (n = 18) to readapt the dissected aortic layers. In all patients, MR imaging was performed on a 1.5-T whole body imaging system for the evaluation of the morphology and function of the heart, aorta and supraaortic branches. RESULTS: Overall hospital mortality following surgical repair of type A aortic dissection was 15.4% during this time period. The highest rate of persistent false lumen perfusion (17/20, 85%) and presence of an intimal flap in the aortic arch (13/20, 65%) was observed in patients following closed repair of acute ascending aortic dissection, whereas the lowest rate of such findings was demonstrated in patients who had undergone open distal aortic repair using biological glue (false lumen perfusion 10/18, 55% and intimal flap in the arch 2/18, 11%). Redo-surgery was significantly reduced in the open repair group using GRF glue (1/18, 5.5%) as compared with the Teflon felt repair group (3/16, 18%) and the closed repair group (6/20, 30%). CONCLUSIONS: In patients with acute type A dissection, open distal aortic repair using GRF-glue favourably influences both (1) the severity of late morphologic alterations in the downstream aorta and (2) the incidence of reoperation. PMID- 10371129 TI - Selective visceral and renal perfusion in thoracoabdominal aneurysm repair. AB - OBJECTIVE: Whether or not selective visceral and renal perfusion during thoracoabdominal aortic aneurysm (TAAA) repair has a protective effect on visceral and renal function remains unknown. The aim of this study was to clarify if selective perfusion has such an effect. METHODS: From May 1982 to December 1997, 82 consecutive patients underwent TAAA repair. Patients receiving hypothermic circulatory arrest or cooling of the kidney using Ringer's lactate solution were excluded, thus 73 patients were enrolled into this study. They were divided into three groups: those in whom selective visceral and renal perfusion was performed using a roller pump (n = 41), those in whom it was performed using a centrifugal pump with a reduced heparin regimen (n = 22) and those who underwent simple aortic clamping alone (n = 10). RESULTS: Serum creatinine, total bilirubin and alanine aminotransferase levels were elevated postoperatively in patients undergoing simple cross-clamp repair, but remained almost within normal limits in patients undergoing TAAA repair with selective visceral and renal perfusion. Urine output was more in selective perfused patients than in non perfused patients. Renal dysfunction, defined by requirement of hemodialysis or by a serum level of creatinine above 3 mg/dl, occurred in four patients (10%) of the roller pump group and in two patients (9%) of the centrifugal pump group, while in four patients (40%) of the simple cross-clamping group. CONCLUSION: Our experience suggests that selective visceral and renal perfusion has a protective effect on hepato-renal function during TAAA repair. PMID- 10371130 TI - Risk factors for intracranial hemorrhage in adults on extracorporeal membrane oxygenation. AB - OBJECTIVE: Intracranial hemorrhage is a recognized complication in neonates and infants on extracorporeal membrane oxygenator support and various risk factors associated with this have been defined. The prevalence and risk factors associated with intracranial hemorrhage in adults on extracorporeal membrane oxygenator support are unknown and this study was performed to define these factors. METHODS: A retrospective study of adults supported with extracorporeal membrane oxygenators at a single institution between January 1992 and December 1996 was performed. Age, gender, weight, body surface area, renal function, anticoagulation, coagulation variables, blood flow, arterial pressure, arterial cannulation sites, duration of support, extracranial bleeding, native cardiac function and presence of intracranial microemboli were analyzed to determine the risk factors for intracranial hemorrhage. RESULTS: Fourteen out of 74 adults on extracorporeal membrane oxygenator support had intracranial hemorrhage (18.9%). An increased risk of intracranial hemorrhage showed a positive correlation with female gender (P = 0.02, odds ratio 6.5), use of heparin (P = 0.05, odds ratio 8.5), creatinine greater than 2.6 mg/ dl (P = 0.009, odds ratio 6.5), need for dialysis (P = 0.03, odds ratio 4.3) and thrombocytopenia (P = 0.007, odds ratio 18.3). Diminishing renal function and the need for dialysis were associated with increasing duration of support. Multivariable logistic regression showed female gender and thrombocytopenia, especially with platelet counts less than 50000 cells/mm3 to be the most important predictors of intracranial hemorrhage. Intracranial hemorrhage was associated with a mortality of 92.3% compared with a mortality of 61% in those without intracranial hemorrhage (P = 0.027). CONCLUSION: Intracranial hemorrhage is a significant complication in adults on extracorporeal membrane oxygenator support. Judicious management of anticoagulation, prevention of renal failure and aggressive correction of thrombocytopenia may help to lower the risk of intracranial hemorrhage in adults on extracorporeal membrane oxygenator support. PMID- 10371131 TI - Induced hypothermia as salvage treatment for refractory cardiac failure following paediatric cardiac surgery. AB - OBJECTIVE: Following corrective cardiac surgery in infants and children for congenital heart disease, a persistent low cardiac output refractory to conventional modes of treatment is associated with a mortality approaching 100%. We advocate the use of whole body hypothermia to reduce tissue oxygen demand and provide a degree of cellular protection against ischaemia allowing time for recovery. We describe our experience. METHODS: Between July 1986 and December 1995, 1885 infants and children underwent surgery (operative mortality, 6%), 1302 requiring cardiopulmonary bypass. Fifty-seven patients had a persistent low cardiac output, impaired respiratory function, decreased urine output and acidosis despite maximal intensive care treatment. Cooling to 32-33 degrees C was therefore started using a thermostatically controlled water filled cooling blanket. RESULTS: Following cooling, there was a fall in heart rate (P<0.001), a rise in mean arterial pressure (P<0.001) and a fall in mean atrial pressure (P<0.001). Significant (P<0.001) increases in pH and urine output were also recorded. Thirty-one (54%) of the 57 patients treated with cooling survived to leave hospital. No long-term sequelae have been noted in these patients. CONCLUSION: Induced hypothermia is a useful salvage treatment, in children following corrective cardiac surgery when all conventional treatment has been tried and failed. PMID- 10371132 TI - Tissue engineering in cardiovascular surgery: MTT, a rapid and reliable quantitative method to assess the optimal human cell seeding on polymeric meshes. AB - OBJECTIVE: Currently used valve substitutes for valve replacement have certain disadvantages that limit their long-term benefits such as poor durability, risks of infection, thromboembolism or rejection. A tissue engineered autologous valve composed of living tissue is expected to overcome these shortcomings with natural existing biological mechanisms for growth, repair, remodeling and development. The aim of the study was to improve cell seeding methods for developing tissue engineered valve tissue. METHODS: Human aortic myofibroblasts were seeded on polyglycolic acid (PGA) meshes. Cell attachment and growth of myofibroblasts on the PGA scaffolds with different seeding intervals were compared to determine an optimal seeding interval. In addition, scanning electron microscopy study of the seeded meshes was also performed to document tissue development. RESULTS: There was a direct correlation between cell numbers assessed by direct counting and MTT(3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltertra-zoliu m bromide) assay. Both attach rate and cell growth seeded on meshes with long intervals (24 and 36 h) were significantly higher than those seeded with short intervals (2 and 12 h) (P<0.01), there was no significant difference between 24- and 36-h seeding interval. Scanning electron microscopy also documented more cell attachment with long seeding intervals resulting in a more solid tissue like structure. CONCLUSION: It is feasible to use human aortic myofibroblasts to develop a new functional tissue in vitro. Twenty-four hours is an optimal seeding interval for seeding human aortic myofibroblasts on PGA scaffolds and MTT test is a rapid and reliable quantitative method to assess the optimal human cell seeding on polymeric meshes. PMID- 10371133 TI - Multicenter results of TADpole heart wire system used to treat postoperative atrial fibrillation. AB - OBJECTIVE: Postoperative atrial fibrillation (AF) affects 20-30% of patients undergoing open-heart surgery, delays mobilization and impairs hemodynamics. Implantation of TADpole Heart Wires offers a new method of applying internal low energy-shocks to terminate AF. The safety and efficacy of the TADpole system to treat postoperative AF was evaluated in this multicenter trial. METHODS: Two atrial wires, configured with a highly flexible 11.5 cm distal shocking and a 0.5 cm proximal pacing electrode were sutured onto the right and left atrium. Upon detection of AF, R-wave synchronized low-energy-shocks were administered via an energy attenuating External Defibrillator Interface Module or ICD programmer. RESULTS: A total of 296 patients (65+/-9.2 years, 74.7% male) have been enrolled to date in six European centers. The wire placement time was 4.2+/-2.2 min, 65 patients had a total of 83 episodes of AF treated by the TADpole Heart Wire system with a conversion rate of 88.5% (approximate energy of 6+/-2 J), early recurrence of AF was observed in ten patients (12.8%). No clinical complications were reported. The shocks were well tolerated with slight sedation. The ease of withdrawal was 2.3+/-1.2 on a graded scale of 0 (easy) to 10 (difficult). CONCLUSIONS: These multicenter results indicate that postoperative atrial cardioversion using TADpole Heart Wires is both safe and efficient. It is expected that hospital length of stay and its associated economic impact can be reduced with this therapy. PMID- 10371134 TI - Pleuro-pericardiocenthesis: an unusual procedure. PMID- 10371135 TI - Both atrial resection and superior vena cava replacement in sleeve pneumonectomy for advanced lung cancer. AB - Extended sleeve pneumonectomy including removal of the superior vena cava, right atrium and parts of left atrium on cardiopulmonary bypass was successfully performed in a 40-year-old man. The tumour was histologically proven a T4 N1 stage with margins free from tumour. Adjuvant radiochemotherapy was administered postoperatively on an outpatient base. The patient did well for 7 months then he died from myocardial infarction due to metastatic infiltration of the right coronary artery. Other metastatic deposits were not found at autopsy. More data from extended pulmonary resections are required to demonstrate a benefit. PMID- 10371136 TI - Mediastinal hibernoma. AB - A 46-year-old asymptomatic male was detected to have a posterior mediastinal mass on a routine check-up. He underwent thoracotomy to remove the mass, which was found to be a hibernoma. The mediastinum is an extremely rare site for an even rarer tumor like the hibernoma. An additional unique feature was the very large tumor size despite which the patient was asymptomatic. Imaging studies are not helpful in revealing its clinically indeterminate nature, hence a surgical specimen is necessary to establish the correct diagnosis. Total excision is advocated for cure, as there is no known malignant potential. PMID- 10371137 TI - Repair of coarctation of the aorta with simultaneous coronary artery bypass grafting without cardiopulmonary bypass. AB - The article describes the management of aortic coarctation associated with coronary artery disease by a one stage surgical procedure without cardiopulmonary bypass in a 44-year-old woman. The vascular prosthesis was anastomosed end-to-end to the descending aorta and a venous bypass was subsequently formed between circumflex coronary artery and the anastomosed vascular prosthesis. There were no postoperative complications. PMID- 10371138 TI - Malignant ventricular arrhythmias revealing anomalous origin of the left coronary artery from the pulmonary artery in two adults. AB - We report two cases of anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA), revealed by malignant ventricular arrhythmias in adult patients. A two coronary system was re-established in both patients, and cryotherapy was performed on one of the patients who, in addition, presented ventricular aneurysm triggering ventricular tachycardia. PMID- 10371139 TI - Congenital subaortic stenosis by accessory mitral valve tissue, recognition and management. AB - Accessory mitral valve tissue as the single cause for left ventricular outflow tract obstruction is a very rare cardiac malformation in normally connected hearts. We report a case in which this condition was present as single cause for left ventricular outflow tract obstruction. The surgical technique is described and a review of the literature presented. PMID- 10371140 TI - Coronary artery spasm after coronary artery bypass grafting. AB - We report a case of a 62-year-old man with severe manifestations of postoperative coronary artery spasm following effective coronary artery bypass grafting. The coronary artery spasm was manifested by ST segment elevation, hypotension and wall motion abnormalities on echocardiography. Urgent angiography confirmed the diagnosis and intracoronary infusion of nitroglycerine and verapamil relieved the coronary spasm. PMID- 10371141 TI - Inferior median sternotomy. PMID- 10371142 TI - Treatment of acute myocardial ischemia during early stages of surgery by an easily applicable method for emergency retroperfusion. PMID- 10371143 TI - Myocardial stunning and dopamine receptor polymorphism. PMID- 10371144 TI - Thermodynamic influences on the fidelity of iron-sulphur cluster formation in proteins. AB - In an organism, two thermodynamic factors are important in ensuring that homometallic [4Fe-4S] cubane clusters are formed in preference to clusters containing heterometals such as Zn or Cu. These are the electronic resonance stabilisation, which boosts the binding of Fe(II) within an Fe-S cluster relative to its normally low position in the Irving-Williams order, and attenuation of the cytoplasmic concentrations of competing metals such as Zn or Cu by specific ligands. PMID- 10371145 TI - Spermine triggers the activation of caspase-3 in a cell-free model of apoptosis. AB - Polyamines are ubiquitous organic cations required for cell proliferation. However, some evidence suggested that their excessive accumulation can induce apoptosis. We show here that, in a post-nuclear extract from U937 cells, the addition of spermine triggers the death program, represented by cytochrome c exit from mitochondria, the dATP-dependent processing of pro-caspase-3 and the onset of caspase activity. Spermine is more effective than spermidine, whereas putrescine has no effect. Polyamine acetylation abolishes their pro-apoptotic power. These data demonstrate a direct mechanism responsible for polyamine toxicity and also suggest that an excessive elevation of free polyamines could be involved in the transduction of a death signal. PMID- 10371146 TI - LIS1 and platelet-activating factor acetylhydrolase (Ib) catalytic subunits, expression in the mouse oocyte and zygote. AB - Platelet-activating factor is a phospholipid with several documented roles in the pre-implantation embryo. Enzymes that belong to the platelet-activating factor acetylhydrolases family inactivate platelet-activating factor. Cytosolic platelet activating factor acetylhydrolase (Ib) is a heterotetramer composed of two catalytic subunits (alpha1/alpha2) and two regulatory LIS1 subunits. The expression of these components was monitored in the mouse oocytes and zygotes using reverse-transcribed PCR and Western blot analysis. Interestingly, these proteins are expressed in the oocyte and zygote and their expression increases after fertilization, probably due to stabilization of maternal RNA. Lis1 mRNA transcription also increases after fertilization. However, assaying for expression of a specific paternal LIS1 isoform detected no zygotic translation in the one cell stage. These findings suggest a potential role for platelet activating factor acetylhydrolase (Ib) components in the early mouse embryo. PMID- 10371147 TI - Major differences in oxysterol formation in human low density lipoproteins (LDLs) oxidized by *OH/O2*- free radicals or by copper. AB - The aim of our study was to determine the oxysterol formation in low density lipoproteins (LDLs) oxidized by defined oxygen free radicals (*OH/O2*-). This was compared to the oxysterol produced upon the classical copper oxidation procedure. The results showed a markedly lower formation of oxysterols induced by *OH/O2*- free radicals than by copper and thus suggested a poor ability of these radicals to initiate cholesterol oxidation in LDLs. Moreover, the molecular species of cholesteryl ester hydroperoxides produced by LDL copper oxidation seemed more labile than those formed upon *OH/O2*(-)-induced oxidation, probably due to their degradation by reaction with copper ions. PMID- 10371148 TI - The interaction between the archaeal elongation factor 1alpha and its nucleotide exchange factor 1beta. AB - In Sulfolobus solfataricus the binding of the exchange factor 1beta (SsEF-1beta) to SsEF-1alpha-GDP displaces the nucleotide and the SsEF-1alpha-SsEF-1beta complex is formed. The complex itself is stable, but it dissociates upon the addition of GDP or Gpp(NH)p but not ATP. Since the rate of the formation of the SsEF-1alpha-SsEF-1beta complex is significatively slower than the rate of the nucleotide exchange catalyzed by SsEF-1beta it can be inferred that in vivo the GDP/GTP exchange reaction proceeds via an SsEF-1alpha-SsEF-1beta interaction without involving the formation of a stable binary complex as an intermediate. PMID- 10371149 TI - Exchange of conductance and gating properties between gap junction hemichannels. AB - Gap junction channels span the membranes of two adjacent cells and allow the gated transit of molecules as large as second messengers from cell to cell. The structure of the gap junction channel pore is not resolved. For identification of pore determinants we used a chimera of two connexins, cx46 and cx32E(1)43, that form membrane channels with distinct unit conductances and channel kinetics. Exchange of the first transmembrane segment (M1) between these connexins resulted in a chimera that exhibited most of the channel properties of the M1 donor, including single channel conductance, channel kinetics, and the preference to dwell at a subconductance level. The M1 segment thus appears to be an important determinant of conductance and gating properties of connexin channels. PMID- 10371150 TI - Multiple tRNA attachment sites in prothymosin alpha. AB - A covalent complex formed by bacterial tRNAs and prothymosin alpha, an abundant acidic nuclear protein involved in proliferation of mammalian cells, upon production of the recombinant rat protein in Escherichia coli cells was studied. Several tRNA attachment sites were identified in the prothymosin alpha molecule using a combination of deletion analysis of prothymosin alpha and site-specific fragmentation of the protein moiety of the prothymosin alpha-tRNA complex. The electrophoretic mobilities of the tRNA-linked prothymosin alpha and its derivatives are consistent with one tRNA molecule attached to one prothymosin alpha molecule, thus suggesting that alternative tRNA linking to one of several available attachment sites occurs. The possible effect of tRNA attachment on the nuclear uptake of prothymosin alpha is discussed. PMID- 10371151 TI - NMR structure of a minimized human agouti related protein prepared by total chemical synthesis. AB - The structure of the chemically synthesized C-terminal region of the human agouti related protein (AGRP) was determined by 2D 1H NMR. Referred to as minimized agouti related protein, MARP is a 46 residue polypeptide containing 10 Cys residues involved in five disulfide bonds that retains the biological activity of full length AGRP. AGRP is a mammalian signaling molecule, involved in weight homeostasis, that causes adult onset obesity when overexpressed in mice. AGRP was originally identified by homology to the agouti protein, another potent signaling molecule involved in obesity disorders in mice. While AGRP's exact mechanism of action is unknown, it has been identified as a competitive antagonist of melanocortin receptors 3 and 4 (MC3r, MC4r), and MC4r in particular is implicated in the hypothalamic control of feeding behavior. Full length agouti and AGRP are only 25% homologous, however, their active C-terminal regions are approximately 40% homologous, with nine out of the 10 Cys residues spatially conserved. Until now, 3D structures have not been available for either agouti, AGRP or their C terminal regions. The NMR structure of MARP reported here can be characterized as three major loops, with four of the five disulfide bridges at the base of the structure. Though its fold is well defined, no canonical secondary structure is identified. While previously reported structural models of the C-terminal region of AGRP were attempted based on Cys homology between AGRP and certain toxin proteins, we find that Cys spacing is not sufficient to correctly determine the 3D fold of the molecule. PMID- 10371152 TI - The vacuolar Ca2+/H+ exchanger Vcx1p/Hum1p tightly controls cytosolic Ca2+ levels in S. cerevisiae. AB - It is well established that the vacuole plays an important role in the cellular adaptation to growth in the presence of elevated extracellular Ca2+ concentrations in Saccharomyces cerevisiae. The Ca2+ ATPase Pmc1p and the Ca2+/H+ exchanger Vcx1p/Hum1p have been shown to facilitate Ca2+ sequestration into the vacuole. However, the distinct physiological roles of these two vacuolar Ca2+ transporters remain uncertain. Here we show that Vcx1p can rapidly sequester a sudden pulse of cytosolic Ca2+ into the vacuole, while Pmc1p carries out this function much less efficiently. This finding is consistent with the postulated role of Vcx1p as a high capacity, low affinity Ca2+ transporter and suggests that Vcx1p may act to attenuate the propagation of Ca2+ signals in this organism. PMID- 10371153 TI - Molecular analysis of a new splice variant of the human melanocortin-1 receptor. AB - The primary hormonal regulator of pigmentation is melanocyte stimulating hormone derived from proopiomelanocortin by proteolytic processing. The melanocortin-1 receptor serves a key role in the regulation of pigmentation. We describe the identification of the first intron within a melanocortin receptor. A new melanocortin-1 receptor isoform, generated by alternative mRNA splicing, encodes an additional 65 amino acids at the predicted intracellular, C-terminal tail of the melanocortin-1 receptor. When expressed in heterologous cells, the new spliced form of the melanocortin-1 receptor (melanocortin-1 receptor B) appears pharmacologically similar to the non-spliced melanocortin-1 receptor. Melanocortin-1 receptor B is expressed in testis, fetal heart and melanomas. PMID- 10371154 TI - Analysis of gene expression data using self-organizing maps. AB - DNA microarray technologies together with rapidly increasing genomic sequence information is leading to an explosion in available gene expression data. Currently there is a great need for efficient methods to analyze and visualize these massive data sets. A self-organizing map (SOM) is an unsupervised neural network learning algorithm which has been successfully used for the analysis and organization of large data files. We have here applied the SOM algorithm to analyze published data of yeast gene expression and show that SOM is an excellent tool for the analysis and visualization of gene expression profiles. PMID- 10371155 TI - Two forms of N intermediate (N(open) and N(closed)) in the bacteriorhodopsin photocycle. AB - Glutaraldehyde, aluminum ions and glycerol (that inhibit the M intermediate decay in the wild-type bacteriorhodopsin and azide-induced M decay in the D96N mutant by stabilization of the M(closed)) accelerate the N decay in the D96N mutant. The aluminum ions, the most potent activator of the N decay, induce a blue shift of the N difference spectrum by approximately 10 nm. Protonated azide as well as acetate and formate inhibit the N decay in both the D96N mutant and the wild-type protein. It is concluded that the N intermediate represents, in fact, an equilibrium mixture of the two ('open' and 'closed') forms. These two forms, like M(closed) and M(open), come to an equilibrium in the microseconds range. The absorption spectrum of the N(open) is slightly shifted to red in comparison to that of the N(closed). Again, this resembles the M forms. 13-cis-all-trans re isomerization is assumed to occur in the N(closed) form only. Binding of 1-2 molecules of protonated azide stabilizes the N(open) form. Existence of the 'open' and 'closed' forms of the M and N intermediates provides the appropriate explanation of the cooperative phenomenon as well as some other effects on the bacteriorhodopsin photocycle. Summarizing the available data, we suggest that M(open) is identical to the M(N) form, whereas M1 and M2 are different substates of M(closed). PMID- 10371156 TI - Reconstruction of the dihydropyridine site in a non-L-type calcium channel: the role of the IS6 segment. AB - Mutations of eight to nine amino acids of IIIS5, IIIS6 and IVS6 segments were shown to reconstruct the dihydropyridine (DHP) interaction site in the non-L-type alpha1E or alpha1A calcium channels. The reconstructed site enabled enantiomer selective inhibition and activation of the expressed chimeras by DHPs but failed to transfer voltage dependence of the current inhibition. Here we show that transfer of four non-conserved amino acids from the IS6 segment to the DHP sensitive alpha1E chimera increased the inhibition by (+)isradipine at the hyperpolarized membrane potential of -100 mV and enhanced the voltage-dependent block. PMID- 10371157 TI - -->H+/2e- stoichiometry in NADH-quinone reductase reactions catalyzed by bovine heart submitochondrial particles. AB - Tightly coupled bovine heart submitochondrial particles treated to activate complex I and to block ubiquinol oxidation were capable of rapid uncoupler sensitive inside-directed proton translocation when a limited amount of NADH was oxidized by the exogenous ubiquinone homologue Q1. External alkalization, internal acidification and NADH oxidation were followed by the rapidly responding (t1/2 < or = 1 s) spectrophotometric technique. Quantitation of the initial rates of NADH oxidation and external H+ decrease resulted in a stoichiometric ratio of 4 H+ vectorially translocated per 1 NADH oxidized at pH 8.0. ADP-ribose, a competitive inhibitor of the NADH binding site decreased the rates of proton translocation and NADH oxidation without affecting -->H+/2e- stoichiometry. Rotenone, piericidin and thermal deactivation of complex I completely prevented NADH-induced proton translocation in the NADH-endogenous ubiquinone reductase reaction. NADH-exogenous Q1 reductase activity was only partially prevented by rotenone. The residual rotenone- (or piericidin-) insensitive NADH-exogenous Q1 reductase activity was found to be coupled with vectorial uncoupler-sensitive proton translocation showing the same -->H+/2e- stoichiometry of 4. It is concluded that the transfer of two electrons from NADH to the Q1-reactive intermediate located before the rotenone-sensitive step is coupled with translocation of 4 H+. PMID- 10371158 TI - Association between the first two immunoglobulin-like domains of the neural cell adhesion molecule N-CAM. AB - The extracellular domain of N-CAM contains five immunoglobulin-like (Ig) and two fibronectin type III-like domains and facilitates cell-cell binding through multiple, weak interdomain interactions. NMR spectroscopy indicated that the two N-terminal Ig-like domains from chicken N-CAM (Ig I and Ig II) interact with millimolar affinity. Physico-chemical studies show that this interaction is significantly amplified when the domains are covalently linked, consistent with an antiparallel domain arrangement. The binding of the two individual domains and the dimerization of the concatenated protein were essentially independent of salt, up to a concentration of 200 mM. The residues in Ig I involved in the interaction map to the BED strands of the beta sandwich, and delineate a largely hydrophobic patch. PMID- 10371159 TI - A cognate dopamine transporter-like activity endogenously expressed in a COS-7 kidney-derived cell line. AB - The activity of the dopamine transporter is an important mechanism for the maintenance of normal dopaminergic homeostasis by rapidly removing dopamine from the synaptic cleft. In kidney-derived COS-7, COS-1 and HEK-293 but not in other mammalian cell lines (CHO, Y1, Ltk-), we have characterized a putative functional dopamine transporter displaying a high affinity (Km approximately 250 nM) and a low capacity (approximately 0.1 pmol/10(5) cells/min) for [3H]dopamine uptake. Uptake displayed a pharmacological profile clearly indicative of the neuronal dopamine transporter. Estimated Ki values of numerous substrates and inhibitors for the COS-dopamine transporter and the cloned human neuronal transporter (human dopamine transporter) correlate well with the exception of a few notable compounds, including the endogenous neurotransmitter dopamine, the dopamine transporter inhibitor GBR 12,909 and the dopaminergic agonist apomorphine. As with native neuronal and cloned dopamine transporters, the uptake velocity was sodium-sensitive and reduced by phorbol ester pre-treatment. Two mRNA species of 3.8 and 4.0 kb in COS-7 cells were revealed by Northern blot analysis similar in size to that seen in native neuronal tissue. A reverse-transcribed PCR analysis confirmed the existence of a processed dopamine transporter. However, no immunoreactive proteins of expected dopamine transporter molecular size or [3H]WIN 35,428 binding activity were detected. A partial cDNA of 1.3 kb, isolated from a COS-1 cDNA library and encoding transmembrane domains 1-6, displayed a deduced amino acid sequence homology of approximately 96% to the human dopamine transporter. Taken together, the data suggest the existence of a non-neuronal endogenous high affinity dopamine uptake system sharing strong functional and molecular homology to that of the cloned neuronal dopamine transporter. PMID- 10371160 TI - Structural and biochemical analysis of Ras-effector signaling via RalGDS. AB - The structure of the complex of Ras with the Ras-binding domain of its effector RalGDS (RGS-RBD), the first genuine Ras-effector complex, has been solved by X ray crystallography. As with the Rap-RafRBD complex (Nasser et al., 1995), the interaction is via an inter-protein beta-sheet between the switch I region of Ras and the second strand of the RGS-RBD sheet, but the details of the interactions in the interface are remarkably different. Mutational studies were performed to investigate the contribution of selected interface residues to the binding affinity. Gel filtration experiments show that the Ras x RGS-RBD complex is a monomer. The results are compared to a recently determined structure of a similar complex using a Ras mutant (Huang et al., 1998) and are discussed in relation to partial loss-of-function mutations and the specificity of Ras versus Rap binding. PMID- 10371161 TI - NMR assignments and secondary structure of the UvrC binding domain of UvrB. AB - The 55 residue C-terminal domain of UvrB that interacts with UvrC during excision repair in Escherichia coli has been expressed and purified as a (His)6 fusion construct. The fragment forms a stable folded domain in solution. Heteronuclear NMR experiments were used to obtain extensive 15N, 13C and 1H NMR assignments. NOESY and chemical shift data showed that the protein comprises two helices from residues 630 to 648 and from 652 to 670. 15N relaxation data also show that the first 11 and last three residues are unstructured. The effective rotational correlation time within the structured region is not consistent with a monomer. This oligomerisation may be relevant to the mode of dimerisation of UvrB with the homologous domain of UvrC. PMID- 10371162 TI - Novel enzymatic oxidation of Mn2+ to Mn3+ catalyzed by a fungal laccase. AB - Fungal laccases are extracellular multinuclear copper-containing oxidases that have been proposed to be involved in ligninolysis and degradation of xenobiotics. Here, we show that an electrophoretically homogenous laccase preparation from the white rot fungus Trametes versicolor oxidized Mn2+ to Mn3+ in the presence of Na pyrophosphate, with a Km value of 186 microM and a Vmax value of 0.11 micromol/min/mg protein at the optimal pH (5.0) and a Na-pyrophosphate concentration of 100 mM. The oxidation of Mn2+ involved concomitant reduction of the laccase type 1 copper site as usual for laccase reactions, thus providing the first evidence that laccase may directly utilize Mn2+ as a substrate. PMID- 10371163 TI - Use of a drug-resistant mutant of stress-activated protein kinase 2a/p38 to validate the in vivo specificity of SB 203580. AB - Stress-activated protein kinase 2a, also called p38, is inhibited by SB 203580 and this drug has been used widely to implicate this enzyme in the regulation of many physiological processes. Here, we introduce a novel method of general application, which can be used to establish whether the effects of SB 203580 are mediated via inhibition of stress-activated protein kinase 2a/p38 or whether they result from 'non-specific' effects. Four events thought to occur upon activation of stress-activated protein kinase 2a/p38 have been established unequivocally. These are the activation of mitogen-activated protein kinase-activated protein kinase-2 and mitogen- and stress-activated protein kinase-1 and the phosphorylation of their presumed substrates, heat shock protein 27 and the transcription factor cyclic AMP response element binding protein, respectively. In contrast, the SB 203580-induced activation of c-Raf is independent of stress activated protein kinase 2a/p38 inhibition. PMID- 10371164 TI - Molecular organization of cholesterol in polyunsaturated phospholipid membranes: a solid state 2H NMR investigation. AB - We compared the molecular organization of equimolar [3alpha-2H1]cholesterol in 18:0-18:1PC (1-stearoyl-2-oleoylphosphatidylcholine), 18:0-22:6PC (1-stearoyl-2 docosahexaenoylphosphatidylcholine), 18:0-20:4PC (1-stearoyl-2 arachidonylphosphatidylcholine) and 20:4-20:4PC (1,2 diarachidonylphosphatidylcholine) bilayers by solid state 2H NMR. Essentially identical quadrupolar splittings (delta v(r) = 45 +/- 1 kHz) corresponding to the same molecular orientation characterized by tilt angle alpha0 = 16 +/- 1 degrees were measured in 18:0-18:1PC, 18:0-22:6PC and 18:0-20:4PC. A profound difference in molecular interaction with dipolyunsaturated 20:4-20:4PC, in contrast, is indicated for the sterol. Specifically, the tilt angle alpha0 = 22 +/- 1 degrees (derived from delta v(r) = 37 +/- 1 kHz) is greater and its membrane intercalation is only 15 mol%. PMID- 10371165 TI - Identification of a novel alternatively spliced septin. AB - Septins are a family of cytoskeletal proteins involved in cytokinesis, targeting of proteins to specific sites on the plasma membrane, and cellular morphogenesis. While many aspects of their function in cytokinesis in yeast cells have been investigated, the function of septins in mammalian cells is less well understood. For example, septins are present in post-mitotic neurons, suggesting they have other roles in, for example, establishing cell polarity. The full extent of the septin gene family is not known in mammalian cells. To better understand the septin gene family, we have cloned and characterized a novel mammalian septin. PMID- 10371166 TI - Characterization of an alternatively spliced isoform of rat vesicle associated membrane protein-2 (VAMP-2). AB - VAMPs are vesicle associated membrane proteins that are essential for secretion. A spliced isoform of rat VAMP-2, called VAMP-2B, is characterized in this study. The VAMP-2B transcript is the result of alternative RNA splicing in which an intron is retained. The predicted amino acid sequence of VAMP-2B differs from VAMP-2 at its carboxy-terminal end. Because recent studies have shown that VAMP's carboxy-terminal end influences the protein's sorting, the location of myc epitope tagged VAMP-2B in PC12 cells was determined. Subcellular fractionation showed colocalization of myc-VAMP-2B and endogenous VAMP-2. Thus alternative RNA splicing does not affect VAMP-2 sorting in PC12 cells. PMID- 10371167 TI - Rate of HIV-1 RNA rebound upon stopping antiretroviral therapy. AB - OBJECTIVE: To determine the rate of plasma HIV-1 RNA rebound in patients stopping highly active antiretroviral therapy (HAART) after achieving undetectable viral load. DESIGN: Sequential plasma HIV RNA levels were measured in six patients during the 21 days following withdrawal from HAART. METHODS: Plasma samples were obtained from six patients who chose to withdraw from HAART because of lipodystrophy, narcotic overdose, insomnia and/or high blood pressure. Longitudinal plasma viral load was determined in triplicate upon stopping therapy. RESULTS: All patients had plasma viral loads below 50 HIV RNA copies/ml at the time of stopping therapy and had had levels below 500 copies/ml for a median of 390 days (range 39-542 days). Plasma HIV rebound upon stopping therapy was rapid (median increase 0.2 log/day; range 0.15-0.42 log/day) and initially appeared to follow first-order kinetics. Plasma HIV RNA levels returned to greater than 500 copies/ml within 6 to 15 days (median 10 days) and approached or exceeded pre-therapy levels in all patients within 21 days of stopping therapy. Extrapolating backwards to the time at which individuals stopped therapy suggested that patients had tens of thousands of total body plasma HIV RNA copies despite having 'undetectable' plasma HIV RNA. CONCLUSIONS: HIV RNA in plasma rebounds within days of stopping antiretroviral therapy. A considerable burden of total body plasma HIV RNA likely remains even during effective HAART therapy. PMID- 10371168 TI - Improvement in neutrophil and monocyte function during highly active antiretroviral treatment of HIV-1-infected patients. AB - OBJECTIVE: To investigate the effect of highly active antiretroviral treatment (HAART) on neutrophil and monocyte function in patients with moderately advanced HIV-1 infection. DESIGN: Eighteen HIV-1-infected patients with CD4 T cell counts below 350/microl, no concomitant active infection, and no previous use of protease inhibitors were treated with indinavir or ritonavir and two reverse transcriptase inhibitors and were followed up for 9 months. Ten age- and sex matched healthy subjects were included as controls. METHODS: The functional activity of neutrophils and monocytes was measured by assessing chemotaxis towards a bacterial peptide, killing activity against Candida albicans, and oxidative burst as measured by chemiluminescence production. RESULTS: Neutrophils and monocytes from the treatment group exhibited a significantly diminished baseline chemotactic and fungicidal activity compared with healthy controls (P < 0.001). After starting HAART, there was a significant improvement in chemotaxis and fungicidal activity of phagocytic cells (P < 0.001). Values of chemotaxis reached normal ranges in 13 out of 18 patients (72%) for neutrophils and eight out of 18 (44%) for monocytes, whereas phagocyte killing was rarely restored to normal values (3/18 cases for monocytes and 0/18 for neutrophils). The administration of HAART was also associated with significantly increased phagocyte chemiluminescence production in response to phorbol-12-myristate 13 acetate or opsonized C. albicans (P < 0.01). CONCLUSION: The functional improvement of two critical components of innate antimicrobial immunity, such as neutrophils and monocytes, may contribute to the improved cell-mediated immune responses against opportunistic infections in HAART-treated patients. PMID- 10371170 TI - Similar rate of disease progression among individuals infected with HIV-1 genetic subtypes A-D. AB - OBJECTIVE: HIV-1 is characterized by a high degree of genetic variation and can be divided into at least 10 distinct genetic subtypes. The purpose of this study was to investigate whether the rate of disease progression shows subtype-specific differences. DESIGN: The investigation was divided into two parts; one study in which 49 ethnic Africans were compared with 49 ethnic Swedes irrespective of the subtype of the infecting virus, and a second study in which 126 individuals infected with different genetic subtypes (28 with subtype A, 59 with subtype B, 21 with subtype C and 18 with subtype D) were compared. METHODS: CD4 cell counts, the rate of CD4 cell decline, plasma HIV-1 RNA levels, clinical status and antiviral treatment were prospectively and retrospectively recorded. The HIV-1 subtype had previously been determined by direct sequencing of the V3 domain of the env gene. RESULTS: There were no significant differences in the rate of CD4 cell decline or clinical disease progression between Africans and Swedes over an observation period of 2 years. Similarly, there were no differences in the rate of CD4 cell decline, clinical progression or plasma HIV-1 RNA levels between individuals infected with subtypes A, B, C or D over a mean observation period of 44 months. CONCLUSION: Neither the genetic subtype of the virus nor the ethnicity of the host appear to be major determinants of disease progression. PMID- 10371169 TI - Expansion of CD57 and CD62L-CD45RA+ CD8 T lymphocytes correlates with reduced viral plasma RNA after primary HIV infection. AB - OBJECTIVE: CD8 T cells, expressing cell surface molecules distinct from those on resting and naive T cells, are increased in HIV infection. The association of increased CD38 and human leukocyte antigen DR (HLA-DR) CD8 T cells with poor prognosis has suggested that activated CD8 T cells may aggravate HIV infection. We examined whether other immunological parameters might influence the viral setpoint. DESIGN: Peripheral T cells from nine untreated patients, obtained after primary HIV infection when plasma HIV had stabilized, were examined for proteins expressed in activated versus resting, memory versus naive, and cytolytic versus non-cytolytic T cells. METHODS: The proportion of CD8 T cells that stain for CD38 and HLA-DR, CD28 and CD57 was compared with plasma viraemia and CD4 cell count. These parameters were also compared with the proportion of CD4 and CD8 T cells that express CD62L and CD45RA, present on naive cells and down-modulated in memory cells. Internal staining for the cytotoxic protein granzyme A was also examined. RESULTS: An increase in CD38 and CD38 HLA-DR CD8 T cells correlated with increased plasma viral RNA (P < 0.00002, P < 0.03, respectively). An increase in CD8 T cells expressing granzyme A was associated with lower CD4 cell counts (P < 0.04). However, the expansion of CD57 and CD62L CD45RA+ CD8 T cells was associated with a lower viral setpoint (P < 0.01, P < 0.02, respectively). CONCLUSION: Phenotypically defined activated CD8 T cells may have different functions in HIV infection. Activated CD8 T cells that are CD57 or CD62L( )CD45RA+ may be beneficial, because their expansion in untreated patients correlates with a reduced viral setpoint after primary infection. PMID- 10371171 TI - The implication of the chemokine receptor CXCR4 in HIV-1 envelope protein-induced apoptosis is independent of the G protein-mediated signalling. AB - OBJECTIVE: The envelope glycoprotein complex (gp120/gp41)n of HIV-1 is one of the viral products responsible for increased apoptosis in HIV infection. Here the role of the chemokine receptor CXCR4 in HIV-1 envelope protein-induced apoptosis was investigated. METHODS: Apoptosis occurring in cocultures of chronically HIV-1 IIIB-infected cells with CD4 target cells expressing the CXCR4 receptor was quantified by terminal deoxinucleotidyl transferase dUTP nick end labeling (TUNEL) or propidium iodide staining followed by fluorescent antibody cell sorting, which allows the evaluation of single-cell killing. Moreover global (single cell- and syncytium-associated) apoptosis was quantified by a new radioactive TUNEL-derived assay. RESULTS: By using these different techniques it was shown that single and syncytium-forming CD4 T cells die by apoptosis upon contact with envelope protein expressing cells independently of viral replication. Moreover, both the CXCR4 agonist SDF-1alpha, and the antagonist AMD3100, showed inhibitory effects on HIV-1 envelope protein-induced apoptosis in the CD4 T-cell subset of peripheral blood mononuclear cells and CD4 cell lines. CXCR4 signalling-induced by HIV-1 envelope proteins in CD4 T cells was not detected. Furthermore, it was shown that envelope protein-induced apoptosis can occur after treating target cells with the Gi-protein inhibitor pertussis toxin. CONCLUSIONS: Evidence is provided for a role of CXCR4 in the mechanisms of HIV envelope protein-induced pathogenesis, contributing to selective CD4 cell killing. The results suggest that CXCR4 is involved in HIV-1-induced apoptosis; however, this role does not appear to involve G-protein-mediated CXCR4 signalling. PMID- 10371172 TI - Incorporation of zidovudine into leukocyte DNA from HIV-1-positive adults and pregnant women, and cord blood from infants exposed in utero. AB - OBJECTIVE: The nucleoside analog 3'-azido-3'-deoxythymidine (ZDV) has widespread clinical use but also is carcinogenic in newborn mice exposed to the drug in utero and becomes incorporated into newborn mouse DNA. This pilot study was designed to determine ZDV incorporation into human blood cell DNA from adults and newborn infants. DESIGN: In this prospective cohort study, peripheral blood mononuclear cells (PBMC) were obtained from 28 non-pregnant adults and 12 pregnant women given ZDV therapy, six non-pregnant adults with no exposure to ZDV, and six non-pregnant adults who last received ZDV > or = 6 months previously. In addition, cord blood leukocytes were obtained from 22 infants of HIV-1-positive, ZDV-exposed women and from 12 infants unexposed to ZDV. There were 11 mother-infant pairs involving HIV-1 -positive women. METHODS: DNA was extracted from PBMC obtained from non-pregnant HIV-1-positive adults taking ZDV, pregnant HIV-1-positive women given ZDV during pregnancy, and from adults not taking ZDV. Cord blood leukocytes were examined from infants exposed to ZDV in utero and from unexposed controls. DNA samples were assayed for ZDV incorporation by anti-ZDV radioimmunoassay (RIA). RESULTS: The majority (76%) of samples from ZDV-exposed individuals, pregnant women (8 of 12), non-pregnant adults (24 of 28), or infants at delivery (15 of 22), had detectable ZDV-DNA levels. The range of positive values for ZDV-treated adults and infants was 25-544 and 22-452 molecules ZDV/10(6) nucleotides, respectively. Analysis of 11 mother-infant pairs showed variable ZDV-DNA incorporation in both, with no correlation by pair or by duration of drug treatment during pregnancy. Two of the 24 samples from individuals designated as controls were positive by anti-ZDV RIA. The 20-fold range for ZDV-DNA values in both adults and infants suggested large interindividual differences in ZDV phosphorylation. CONCLUSIONS: Incorporation of ZDV into DNA was detected in most of the samples from ZDV-exposed adults and infants. Therefore, the biologic significance of ZDV-DNA damage and potential subsequent events, such as mutagenicity, should be PMID- 10371173 TI - Rapid disease progression in HIV-1 perinatally infected children born to mothers receiving zidovudine monotherapy during pregnancy. The Italian register for HIV Infection in Children. AB - OBJECTIVE: To investigate the outcome in children perinatally infected with HIV-1 whose mothers received zidovudine (ZDV) monotherapy in pregnancy. DESIGN: Observational retrospective study of a prospectively recruited cohort. SETTING: Italian Register for HIV Infection in Children. PATIENTS: A group of 216 children perinatally infected with HIV-1, born in 1992-1997 and derived prospectively from birth: 38 children had mothers receiving ZDV monotherapy and for 178 children the mothers received no antiretroviral treatment during pregnancy. MAIN OUTCOME MEASURES: The estimated probability of developing severe disease or severe immune suppression, survival probability [95% confidence interval (CI)] within 3 years, and the hazard ratio (95% CI), adjusted for year of birth, maternal clinical condition at delivery, birthweight and treatments (Pneumocystis carinii pneumonia chemoprophylaxis and/or antiretroviral therapy before the onset of severe disease, severe immune suppression or death) were compared. RESULTS: Comparison of HIV-1-infected children whose mothers were treated with ZDV with children whose mothers were not treated showed that the former group had a higher probability of developing severe disease [57.3% (95% CI 40.9-74.3) versus 37.2% (95% CI 30.0-45.4); log-rank test 7.83, P = 0.005; adjusted hazard ratio 1.8 (95% CI 1.1-3.1)] or severe immune suppression [53.9% (95% CI 36.3-73.5) versus 37.5% (95% CI 30.0-46.2); log-rank test 5.58, P = 0.018; adjusted hazard ratio 2.4, (95% CI: 1.3-4.3)] and a lower survival [72.2% (95% CI 50.4-85.7) versus 81.0% (95% CI 73.7-86.5); log-rank test 4.23, P = 0.039; adjusted hazard ratio of death 1.9 (95% CI 1.1-3.6)]. CONCLUSIONS: This epidemiological observation could stimulate virologic studies to elucidate whether this rapid progression depends on in utero infection or transmission of resistant virus. Findings may suggest a need to hasten HIV-1 diagnosis in infants of ZDV-treated mothers and undertake an aggressive antiretroviral therapy in those found to be infected. PMID- 10371174 TI - Dynamics of plasma cytokine levels in patients with advanced HIV infection and active tuberculosis: implications for early recognition of patients with poor response to anti-tuberculosis treatment. AB - OBJECTIVE: To examine whether the serial measurement of plasma cytokine levels can assist in the early recognition of AIDS/tuberculosis patients with poor response to anti-tuberculosis treatment. DESIGN: Longitudinal, prospective cohort study. SETTING: A university hospital, the largest centre for HIV/AIDS patients in Taiwan. METHODS: Between January 1997 and September 1998, 25 consecutive patients with advanced HIV infection and suspected tuberculosis were enrolled in the study. Plasma samples were obtained on day 1 (baseline), 3, 7 and 14 of anti tuberculosis treatment and the levels of tumour necrosis factor-alpha (TNF-alpha) were measured. Patients were classified as either responders or non-responders according to the results of assessment of symptoms and follow-up cultures during the sixth and eighth week of anti-tuberculosis treatment. Thirty consecutive HIV negative tuberculosis patients were also enrolled in the study. RESULTS: The data of a total of 16 AIDS patients (median CD4 cell count 16 x 10(6)/l; 12 responders and four non-responders) and 21 HIV-negative patients (16 responders and five non responders), whose tuberculosis was culture-proven, were included for analysis. In responders, TNF-alpha levels declined remarkably within the first week of anti tuberculosis treatment; however, the decline of TNF-alpha levels in non responders was significantly less [the median ratio of TNF-alpha level on day 7 to that at baseline was 0.32 versus 0.85 (P < 0.001) in AIDS patients; 0.34 versus 0.80 (P = 0.001) in HIV-negative patients). The lack of a > or = 50% reduction in pre-treatment TNF-alpha levels during the first week of treatment was strongly associated with a poor response to anti-tuberculosis treatment (P = 0.001 in AIDS patients; P < 0.001 in HIV-negative patients). CONCLUSION: Serial measurement of plasma TNF-alpha levels may help to assess the response to anti tuberculosis treatment in AIDS patients, in spite of very low CD4 cell counts. Failure of TNF-alpha levels to decline by > or = 50 % of pre-treatment levels in the first week of treatment may be an early surrogate marker of a poor response. PMID- 10371175 TI - Anaemia is an independent predictive marker for clinical prognosis in HIV infected patients from across Europe. EuroSIDA study group. AB - OBJECTIVES: To describe changes in haemoglobin over time and to determine the joint prognostic value of the current haemoglobin, CD4 lymphocyte count and viral load among patients from across Europe. PATIENTS: The analysis included 6725 patients from EuroSIDA, an observational, prospective cohort of patients with HIV from across Europe. METHODS: Normal haemoglobin was defined as haemoglobin greater than 14 g/dl for men and 12 g/dl for women; mild anaemia was 8-14 g/dl for men and 8-12 g/dl for women; severe anaemia was defined as less than 8 g/dl for both males and females. Linear regression techniques were used to estimate the annual change in haemoglobin; standard survival techniques were used to describe disease progression and risk of death. RESULTS: At recruitment to the study, 40.4% had normal levels of haemoglobin, 58.2% had mild anaemia and 1.4% had severe anaemia. At 12 months after recruitment, the proportion of patients estimated to have died was 3.1% [95% confidence interval (CI) 2.3-3.9] for patients without anaemia, 15.9% for patients with mild anaemia (95% CI 14.5-17.2) and 40.8% for patients with severe anaemia (95% CI 27.9-53.6; P < 0.0001). In a multivariate, time-updated Cox proportional hazards model, adjusted for demographic factors, AIDS status and each antiretroviral treatment as time dependent covariates, a 1 g/dl decrease in the latest haemoglobin level increased the hazard of death by 57% [relative hazard (RH) 1.57; 95% CI 1.41-1.75; P < 0.0001], a 50% drop in the most recent CD4 lymphocyte count increased the hazard by 51% (RH 1.51; 95% CI 1.35-1.70; P < 0.0001) and a log increase in the latest viral load increased the hazard by 37% (RH 1.37; 95% CI 1.15-1.63; P = 0.0005). CONCLUSIONS: Severe anaemia occurred infrequently among these patients but was associated with a much faster rate of disease progression. Among patients with similar CD4 lymphocyte counts and viral load, the latest value of haemoglobin was a strong independent prognostic marker for death. PMID- 10371176 TI - Determinants of sustainable CD4 lymphocyte count increases in response to antiretroviral therapy. AB - OBJECTIVE: HIV-induced CD4 lymphocyte depletion is partially reversed by antiretroviral therapy but it is unclear if the degree to which the CD4 count rises depends on viral suppression (if so, the extent of viral suppression required to achieve a maximal CD4 count rise), whether the rise is sustainable and whether it occurs in patients with CD4 count <10 x 10(6) cells/l. We aimed to address these issues. METHODS: We studied CD4 count and plasma HIV RNA values every 4 weeks for 72 weeks in 154 patients starting indinavir-containing regimens. RESULTS: Mean baseline HIV RNA and CD4 count were 4.8 log10 copies/ml and 180 x 10(6) cells/l, respectively. Overall, there was a mean increase in CD4 count of 143 x 10(6) cells/l by 72 weeks. The adjusted mean increase (adjusted for initial viral load, CD4 count and age) was strongly related to the mean viral suppression over the follow-up period (P < 0.0001). Importantly, there was a highly significant difference (P = 0.0004) in the rise in CD4 count between those with 2-3 log suppression (161 x 10(6) cells/l) and those with > 3 log suppression (314 x 10(6) cells/l; mean 3.6 log suppression in this group), suggesting that with even greater suppression the rise in CD4 lymphocytes may be still larger. We also studied whether CD4 counts were still rising after 72 weeks in patients with sustained suppression of at least 3 log in viral load. There was a significant (P = 0.004; paired t-test) rise in count of 43 x 10(6) cells/l between weeks 64 and 72 in these patients, suggesting that regeneration continues at least up to 72 weeks after therapy, provided virus replication continues to be suppressed. Patients with initial CD4 counts < 10 x 10(6) cells/l experienced no smaller rises than those at higher levels, even after adjustment for other factors. CONCLUSION: These results strongly support a direct causal relationship between HIV replication and CD4 lymphocyte count depletion. The rise in those with > 3 log suppression provides the best available indicator of the potential for natural CD4 regeneration in HIV-infected patients. However, since still greater CD4 count rises may be seen with more suppressive regimens, it may not be possible to study the intrinsic CD4 regenerative capacity until such regimens are available. PMID- 10371177 TI - Nevirapine induced opiate withdrawal among injection drug users with HIV infection receiving methadone. AB - BACKGROUND: Pharmacokinetic interactions complicate and potentially compromise the use of antiretroviral and other HIV therapeutic agents in patients with HIV disease. This may be particularly so among those receiving treatment for substance abuse. OBJECTIVE: We describe seven cases of opiate withdrawal among patients receiving chronic methadone maintenance therapy following initiation of therapy with the non-nucleoside reverse transcriptase inhibitor, nevirapine. DESIGN: Retrospective chart review. RESULTS: In all seven patients, due to the lack of prior information regarding a significant pharmacokinetic interaction between these agents, the possibility of opiate withdrawal was not anticipated. Three patients, for whom methadone levels were available at the time of development of opiate withdrawal symptoms, had subtherapeutic methadone levels. In each case, a marked escalation in methadone dose was required to counteract the development of withdrawal symptoms and allow continuation of antiretroviral therapy. Three patients continued nevirapine with methadone administered at an increased dose; however, four chose to discontinue nevirapine. CONCLUSION: To maximize HIV therapeutic benefit among opiate users, information is needed about pharmacokinetic interactions between antiretrovirals and therapies for substance abuse. PMID- 10371178 TI - Costs of HIV medical care in the era of highly active antiretroviral therapy. AB - OBJECTIVES: In the USA, Medicaid is the principal payer of the health care costs of patients with HIV infection. We wished to determine how the costs to Medicaid of patients in Maryland infected with HIV have changed in the setting of highly active antiretroviral treatment. DESIGN: Observational cohort study. METHODS: Analysis of combined economic and clinical data of patients from the Johns Hopkins HIV Service, the provider of primary and sub-specialty care for a majority of HIV-infected patients in the Baltimore metropolitan region. All patients were enrolled in Medicaid and received care longitudinally in Maryland from 1 January 1995 through 31 December 1997. Monthly Medicaid payments were calculated for all inpatient and outpatient services by fiscal year, CD4 cell count, and use of protease inhibitors. RESULTS: For inpatients with a CD4 cell count < or = 50 x 10(6) cells/l, the total health care average monthly payments remained unchanged ($2629 in 1995, $2585 in 1997). Total mean monthly payments increased for those with a CD4 cell count > 50 x 10(6) cells/l (CD4 cell count 50 200 x 10(6) cells/l, $1172 in 1995 and $1615 in 1997, P < 0.05; CD4 cell count 201-500 x 10(6) cells/l, $1078 in 1995 and $1305 in 1997, P < 0.05). However, when data were stratified according to use of a protease inhibitor-containing regimen (used during approximately 50% of follow-up time in 1996-1997) it was found that hospital inpatient payments decreased significantly in all CD4 strata for patients on a protease inhibitor-containing regimen whereas pharmacy payments increased significantly. Inpatient payments associated with treating opportunistic illness were lower in 1996-1997 for patients receiving protease inhibitor therapy compared with those not receiving protease inhibitors. On balance, total health care payments tended to be slightly lower for patients receiving a protease inhibitor regimen. CONCLUSION: Although protease inhibitor containing antiretroviral regimens are being used by only about half of our Medicaid-insured patients, when they are used, there are significantly lower hospital inpatient and community care costs, as well as lower costs associated with the treatment of opportunistic illness. Even with the concurrent increase in their pharmacy costs, total health care costs were stable or slightly lower for these patients. We believe this is a favorable result suggesting a good clinical value being achieved without an increase in costs. PMID- 10371179 TI - Gonorrhea incidence and HIV testing and counseling among adolescents and young adults seen at a clinic for sexually transmitted diseases. AB - OBJECTIVE: To determine whether HIV testing and posttest counseling may be associated with an increase in gonorrhea incidence among adolescents and young adults seen at a clinic for sexually transmitted diseases (STD). DESIGN: A historical cohort study with the collection of longitudinal data on the patients' HIV testing and counseling experience. SETTING: Delgado STD clinic of New Orleans, Louisiana, a public ambulatory primary care center that serves mainly the economically disadvantaged Black population. PATIENTS: A record-based inception cohort of 4031 patients aged 15-25 years diagnosed at the clinic between June 1989 and May 1991 with a first lifetime gonorrhea infection. INTERVENTION: Routine confidential HIV tests and posttest counseling sessions experienced at the clinic during follow-up. OUTCOME MEASURE: Incidence rate of reported gonorrhea reinfection. RESULTS: Of the patients, 51.5% were tested once for HIV antibodies and 25.9% twice or more. Formal posttest counseling occurred after 8.5% of the 4665 HIV-negative and 44.0% of the 49 HIV-positive tests. In the most pessimistic of several models controlling for history of gonorrhea, HIV testing and counseling history, and other potential confounding factors, a significantly lower rate of gonorrhea reinfection was observed after a first HIV negative test than before [adjusted relative risk (RR), 0.66; 95% confidence interval (CI), 0.59-0.74; P < 0.00011. As compared with the pretest period, significantly higher rates of gonorrhea were observed after respectively a second (RR, 1.18; 95% CI, 1.01-1.37; P = 0.03) and a third (RR, 1.52; 95% CI, 1.22-1.88; P = 0.0001) HIV-negative test. No significant association was found between HIV positive testing and any variation in gonorrhea rate (RR, 0.95; 95% CI, 0.56 1.62; P = 0.85). Posttest counseling for HIV-negative and HIV-positive results were followed respectively by a significantly higher rate of gonorrhea (RR; 1.27; 95% CI, 1.09-1.48; P = 0.002) and a non-significantly lower rate of gonorrhea (RR, 0.53; 95% CI, 0.17-1.60; P = 0.85). CONCLUSION: Our results do not exclude the possibility of a modest increase in gonorrhea incidence after routine HIV testing and counseling in an STD clinic. Nevertheless, this conclusion holds only under the least favorable assumptions and applies solely to a minority of patients. PMID- 10371180 TI - Prevalence of primary HIV drug resistance among seroconverters during an explosive outbreak of HIV infection among injecting drug users. AB - OBJECTIVES: This study examined the frequency of transmission of drug resistant HIV in the population of injecting drug users (IDU) in Vancouver, Canada during a period of particularly high virus transmission. DESIGN: All subjects enrolled in the Vancouver Injection Drug Users Study who seroconverted from HIV negative to positive status (n = 61) between December 1996 and February 1998 were eligible for analysis. The first seropositive sample from 57 individuals with plasma samples available was analyzed for resistance to antiretroviral agents by population based sequencing of the HIV protease and reverse transcriptase genes. METHODS: Plasma viral RNA was extracted and the viral reverse transcriptase and protease regions were amplified by nested reverse transcription-PCR. The presence of mutations associated with antiretroviral drug resistance was assessed by automated sequence analysis. RESULTS: Protease and reverse transcriptase sequences were successfully obtained from the 57 recent seroconverters. No cases of transmission of variants associated with significant resistance to protease inhibitors or nucleoside and non-nucleosides reverse transcriptase inhibitors were detected. CONCLUSION: The frequency of transmission of drug resistant HIV amongst these recently infected IDU is extremely low, with no protease or reverse transcriptase inhibitor resistant strains detected soon after seroconversion. The data provide no rationale for withholding treatment from this already marginalized population. PMID- 10371181 TI - Prevalence of unprotected sex among men with AIDS in Los Angeles County, California 1995-1997. AB - OBJECTIVE: To determine the prevalence of unprotected sex among men with AIDS in Los Angeles County. DESIGN: Cross-sectional study. METHODS: All men aged > or = 18 years who were newly reported to the local health department with AIDS and completed a standardized interview between January 1995 and June 1997 were included in the study. Men were classified as having unprotected sex if they reported one or more sex partners during the past year with whom they had vaginal or anal sex and did not always use a condom. RESULTS: Of 617 men interviewed, 29% reported unprotected sex in the past year. The prevalence of unprotected sex was highest among men < 30 years of age (43%) and those who had first learned of their HIV-positive status < 12 months prior to interview (44%). In all, 323 (52%) men reported one or more male sex partners in the past year. Of these, 22% reported unprotected insertive anal sex and 27% unprotected receptive anal sex. One or more female partners in the past year was reported by 131 (21%) men. Of these, 53% reported unprotected vaginal sex and 18% unprotected anal sex. CONCLUSIONS: The findings highlight the importance of early HIV detection efforts, coupled with targeted and sustained HIV prevention services for those who test positive, to prevent ongoing transmission of the virus. PMID- 10371182 TI - Reasons for failure of prophylaxis for Pneumocystis carinii pneumonia. PMID- 10371184 TI - Dichotomous effects of macrophage-derived chemokine on HIV infection. PMID- 10371183 TI - Detection of multiple drug-resistance-associated pol mutations in cervicovaginal secretions from women largely treated with antiretroviral agents. PMID- 10371185 TI - HIV-negative infants born to HIV-1 but not HIV-2-positive mothers fail to develop a Bacillus Calmette-Guerin scar. PMID- 10371186 TI - Higher antiviral activity of antiretroviral regimens including protease inhibitors. PMID- 10371187 TI - Anaphylaxis after rechallenge with abacavir. PMID- 10371188 TI - Effects of metformin on insulin resistance and central adiposity in patients receiving effective protease inhibitor therapy. PMID- 10371190 TI - Nevirapine-induced withdrawal symptoms in HIV patients on methadone maintenance programme: an alert. PMID- 10371189 TI - Lamivudine for chronic hepatitis B and HIV co-infection. PMID- 10371191 TI - Decreased needs for hospital care and antibiotics in children with advanced HIV-1 disease after protease inhibitor-containing combination therapy. PMID- 10371192 TI - Tumour necrosis factor, a key role in obesity? AB - Tumour necrosis factor-alpha (TNF) is a pleiotropic cytokine involved in many metabolic responses in both normal and pathophysiological states. In spite of the fact that this cytokine (also known as "cachectin") has been related to many of the metabolic abnormalities associated with cachexia, recent studies suggest that TNF may also have a central role in obesity modulating energy expenditure, fat deposition and insulin resistance. This review deals with the role of TNF in the control of fat mass and obesity. PMID- 10371193 TI - Characterisation of a plant 3-phosphoinositide-dependent protein kinase-1 homologue which contains a pleckstrin homology domain. AB - A plant homologue of mammalian 3-phosphoinositide-dependent protein kinase-1 (PDK1) has been identified in Arabidopsis and rice which displays 40% overall identity with human 3-phosphoinositide-dependent protein kinase-1. Like the mammalian 3-phosphoinositide-dependent protein kinase-1, Arabidopsis 3 phosphoinositide-dependent protein kinase-1 and rice 3-phosphoinositide-dependent protein kinase-1 possess a kinase domain at N-termini and a pleckstrin homology domain at their C-termini. Arabidopsis 3-phosphoinositide-dependent protein kinase-1 can rescue lethality in Saccharomyces cerevisiae caused by disruption of the genes encoding yeast 3-phosphoinositide-dependent protein kinase-1 homologues. Arabidopsis 3-phosphoinositide-dependent protein kinase-1 interacts via its pleckstrin homology domain with phosphatidic acid, PtdIns3P, PtdIns(3,4,5)P3 and PtdIns(3,4)P2 and to a lesser extent with PtdIns(4,5)P2 and PtdIns4P. Arabidopsis 3-phosphoinositide-dependent protein kinase-1 is able to activate human protein kinase B alpha (PKB/AKT) in the presence of PtdIns(3,4,5)P3. Arabidopsis 3-phosphoinositide-dependent protein kinase-1 is only the second plant protein reported to possess a pleckstrin homology domain and the first plant protein shown to bind 3-phosphoinositides. PMID- 10371194 TI - Isolation of reductase for SoxR that governs an oxidative response regulon from Escherichia coli. AB - SoxR is a transcription factor triggered by oxidative stress in Escherichia coli. Recent evidence suggests that novel redox regulation couples oxidation state to promoter activation. We have isolated the reductase for SoxR in E. coli using an assay of NADPH- and SoxR-dependent cytochrome c reductase activity. When the purified protein was incubated in an anaerobic reaction mixture containing SoxR and NADPH, the reduction of [2Fe-2S] cluster of SoxR was observed by optical and EPR spectroscopy. Our results indicate that the purified protein serves as an NADPH-dependent reduction system for SoxR. PMID- 10371195 TI - Addressing substrate glutamine requirements for tissue transglutaminase using substance P analogues. AB - We have investigated the effect on the substrate requirements for guinea pig liver (tissue) transglutaminase of a set of 11 synthetic glutamine substitution analogues making up the full sequence of the naturally occurring tissue transglutaminase substrate substance P. While a number of peptide sequences derived from proteins that are well-recognized as tissue transglutaminase substrates have been studied, the enzyme activity using substitution analogues of full-length natural substrates has not been investigated as thoroughly. Thus, our set of substance P analogues only differs from one to other by one amino acid mutation while the length (of the peptide) is maintained as in the natural parent peptide. Our results indicate that a glutamine residue is not recognized as substrate by the enzyme whether it is placed at the N- or C-terminal or between two positively charged residues or between two proline residues. To further address the effect on enzyme activity of charged amino acids in the vicinity of the reactive glutamine residue, a new set of synthetic charge replacement analogues of substance P has been also studied. Together, the results have identified new minimal requirements for modification of a particular glutamine residue in a polypeptide chain. It would be of interest to set up a full set of such requirements in order to highlight potential glutamine residues as enzyme targets in the growing list of proteins that are being described as transglutaminase substrates. PMID- 10371196 TI - Human milk lactoferrin binds two DNA molecules with different affinities. AB - Evidence is presented that lactoferrin (LF), an Fe3+-binding glycoprotein, possesses two DNA-binding sites with different affinities for specific oligonucleotides (ODNs) (Kdl = 8 nM; Kd2 approximately 0.1 mM). The high affinity site became labeled after incubation with affinity probes for DNA-binding sites; like the antibacterial and polyanion-binding sites, this site was shown to be located in the N-terminal domain of LF. Interaction of heparin with the polyanion binding site inhibits the binding of ODNs to both sites. These data suggest that the DNA-binding sites of LF coincide or overlap with the known polyanion and antimicrobial domains of the protein. PMID- 10371197 TI - Binding of amyloid beta-peptide to mitochondrial hydroxyacyl-CoA dehydrogenase (ERAB): regulation of an SDR enzyme activity with implications for apoptosis in Alzheimer's disease. AB - The intracellular amyloid beta-peptide (A beta) binding protein, ERAB, a member of the short-chain dehydrogenase/reductase (SDR) family, is known to mediate apoptosis in different cell lines and to be a class II hydroxyacyl-CoA dehydrogenase. The A beta peptide inhibits the enzymatic reaction in a mixed type fashion with a Ki of 1.2 micromol/l and a KiES of 0.3 micromol/l, using 3 hydroxybutyryl-CoA. The peptide region necessary for inhibition comprises residues 12-24 of A beta1-40, covering the 16-20 fragment, which is the minimum sequence for the blockade of A beta polymerization, but that minimal fragment is not sufficient for more than marginal inhibition. The localization of ERAB to the endoplasmic reticulum and mitochondria suggests a complex interaction with components of the programmed cell death machinery. The interaction of A beta with ERAB further links oxidoreductase activity with both apoptosis and amyloid toxicity. PMID- 10371198 TI - Targeting of tail-anchored proteins to yeast mitochondria in vivo. AB - Tail-anchored proteins are inserted into intracellular membranes via a C-terminal transmembrane domain. The topology of the protein is such that insertion must occur post-translationally, since the insertion sequence is not available for membrane insertion until after translation of the tail-anchored polypeptide is completed. Here, we show that the targeting information in one such tail-anchored protein, translocase in the outer mitochondrial membrane 22, is contained in a short region flanking the transmembrane domain. An equivalent region is sufficient to specify the localisation of Bcl2 and SNARE proteins to the secretory membranes. We discuss the targeting process for directing members of this protein family to the secretory and mitochondrial membranes in vivo. PMID- 10371199 TI - The origin of cecropins; implications from synthetic peptides derived from ribosomal protein L1. AB - We recently showed that Helicobacter pylori grown on plates produce cecropin-like antibacterial peptides to which H. pylori is resistant. This antibacterial activity was traced to fragments from the N-terminus of ribosomal protein L1 (Putsep et al., Nature, April 22, 1999). The evolutionary suggestion from this finding has now been extended by the synthesis of eight peptides with sequences taken from the N-terminus of ribosomal protein L1 (RpL1) of five different species. Two peptides of different length derived from H. pylori RpL1 showed a potent antibacterial activity, while a peptide with the sequence from Escherichia coli was 20 times less active. Like cecropins the H. pylori peptides were not cytolytic. We suggest that the cecropins have evolved from ribosomal protein L1 of an ancestral intracellular pathogen that developed to a symbiont ending as an organelle. When the R1 gene moved into the host nucleus, a duplication provided a copy from which today cecropins could have evolved. PMID- 10371200 TI - Evolutionary divergence in the broad complex, tramtrack and bric a brac/poxviruses and zinc finger domain from the candidate tumor suppressor gene hypermethylated in cancer. AB - Hypermethylated in cancer, a new candidate tumor suppressor gene located in 17p13.3, encodes a protein with five Kruppel-like C2H2 zinc finger motifs and a N terminal protein/protein interaction domain called broad complex, tramtrack and bric a brac/poxviruses and zinc finger domain. Hypermethylated in cancer appears unique in the broad complex, tramtrack and bric a brac/poxviruses and zinc finger family since it contains a 13 amino acid insertion located in a loop between the conserved beta-strand beta5 and helix alpha5 which are involved in dimerization and scaffolding of the broad complex, tramtrack and bric a brac/poxviruses and zinc finger domain. Cloning and sequencing of a murine hypermethylated in cancer gene suggests that this insertion has been acquired late in the evolution since it is present in two mammalian hypermethylated in cancer genes but absent in its zebrafish and avian counterparts. This is a unique example of a high divergence of the same broad complex, tramtrack and bric a brac/poxviruses and zinc finger domain in different species. PMID- 10371202 TI - Modified peptide nucleic acids are internalized in mouse macrophages RAW 264.7 and inhibit inducible nitric oxide synthase. AB - Overexpression of inducible nitric oxide synthase causes the production of high levels of nitric oxide, which, under pathological conditions, leads to immunosuppression and tissue damage. The results recently obtained using peptide nucleic acids, rather than traditional oligonucleotides as antigen and antisense molecules, prompted us to test their efficacy in the regulation of nitric oxide production, thereby overcoming the obstacle of cellular internalization. The cellular permeability of four inducible nitric oxide synthase antisense peptide nucleic acids of different lengths was evaluated. These peptide nucleic acids were covalently linked to a hydrophobic peptide moiety to increase internalization and to a tyrosine to allow selective 125I radiolabelling. Internalization experiments showed a 3-25-fold increase in the membrane permeability of the modified peptide nucleic acids with respect to controls. Inducible nitric oxide synthase inhibition experiments on intact stimulated macrophages RAW 264.7 after passive permeation of the two antisense peptide nucleic acids 3 and 4 demonstrated a significant decrease (43-44%) in protein enzymatic activity with respect to the controls. These data offer a basis for developing a good alternative to conventional drugs directed against inducible nitric oxide synthase overexpression. PMID- 10371201 TI - Ca2+-dependent interaction of the trpl cation channel and calmodulin. AB - The transient receptor potential-like ion channel from Drosophila melanogaster was originally identified as a calmodulin binding protein (Philips et al., 1992) involved in the dipterian phototransduction process. We used a series of fusion proteins and an epitope expression library of transient receptor potential-like fusion proteins to characterize calmodulin binding regions in the transient receptor potential-like channel through the use of [125I]calmodulin and biotinylated calmodulin and identified two distinct sites at the C-terminus of the transient receptor potential-like ion channel. Calmodulin binding site 1, predicted from searching of the primary structure for amphiphilic helices (Philips et al., 1992), covers a 16 amino acid sequence (S710-I725) and could only be detected through biotinylated calmodulin. Calmodulin binding site 2 comprises at least 13 amino acids (K859ETAKERFQRVAR871) and binds both [125I]calmodulin and biotinylated calmodulin. Both sites (i) bind calmodulin at least in a one to one stoichiometry, (ii) differ in their affinity for calmodulin revealing apparent Ki values of 12.3 nM (calmodulin binding site 1) and 1.7 nM (calmodulin binding site 2), respectively, (iii) bind calmodulin only in the presence of Ca2+ with 50% of site 1 and site 2, respectively, occupied by calmodulin in the presence of 0.1 microM (calmodulin binding site 1) and 3.3 microM Ca2+ (calmodulin binding site 2) and give evidence that (iv) a Ca2+ calmodulin-dependent mechanism contributes to transient receptor potential-like cation channel modulation when expressed in CHO cells. PMID- 10371203 TI - Inhibition of amylase secretion from differentiated AR4-2J pancreatic acinar cells by an actin cytoskeleton controlled protein tyrosine phosphatase activity. AB - Disruption of the actin cytoskeleton in AR4-2J pancreatic acinar cells led to an increase in cytosolic protein tyrosine phosphatase activity, abolished bombesin induced tyrosine phosphorylation and reduced bombesin-induced amylase secretion by about 45%. Furthermore, both tyrosine phosphorylation and amylase secretion induced by phorbol ester-induced activation of protein kinase C were abolished. An increase in the cytosolic free Ca2+ concentration by the Ca2+ ionophore A23187 had no effect on tyrosine phosphorylation but induced amylase release. Only when added together with phorbol ester, the same level of amylase secretion as with bombesin was reached. This amylase secretion was inhibited by about 40%, by actin cytoskeleton disruption similar to that induced by bombesin. We conclude that actin cytoskeleton-controlled protein tyrosine phosphatase activity downstream of protein kinase C activity regulates tyrosine phosphorylation which in part is involved in bombesin-stimulated amylase secretion. PMID- 10371204 TI - Circadian-regulated expression of a nuclear-encoded plastid sigma factor gene (sigA) in wheat seedlings. AB - The activity of a light-responsive psbD promoter in plastids is known to be regulated by a circadian clock. However, the mechanism of the circadian regulation of the psbD light-responsive promotor, which is recognized by an Escherichia coli-type RNA polymerase, is not yet known. We examined the time course of mRNA accumulation of two E. coli-type RNA polymerase subunit genes, sigA and rpoA, under a continuous light condition after 12 h light/12 h dark entrainment. Accumulation of the sigA mRNA was found to be regulated by a circadian clock, while rpoA mRNA did not show any significant oscillation throughout the experiment. PMID- 10371205 TI - Antisense oligonucleotides against receptor kinase GRK2 disrupt target selectivity of beta-adrenergic receptors in atrial myocytes. AB - K+ channels composed of GIRK subunits are predominantly expressed in the heart and various regions of the brain. They are activated by betagamma-subunits released from pertussis toxin-sensitive G-proteins coupled to different seven helix receptors. In rat atrial myocytes, activation of K(ACh) channels is strictly limited to receptors coupled to pertussis toxin-sensitive G-proteins. Upon treatment of myocytes with antisense oligodesoxynucleotides against GRK2, a receptor kinase with Gbetagamma binding sites, in a fraction of cells, K(ACh) channels can be activated by beta-adrenergic receptors. Sensitivity to beta agonist is insensitive to pertussis toxin treatment. These findings demonstrate a potential role of Gbetagamma binding proteins for target selectivity of G-protein coupled receptors. PMID- 10371206 TI - Oncogenic Ras-induced germinal vesicle breakdown is independent of phosphatidylinositol 3-kinase in Xenopus oocytes. AB - A number of reports have identified phosphatidylinositol 3-kinase as a downstream effector of Ras in various cellular settings, in contrast to others supporting the notion that phosphatidylinositol 3-kinase acts upstream of Ras. Here, we used Xenopus oocytes, a model of Ras-mediated cell cycle progression (G2/M transition) to analyze the contribution of phosphatidylinositol 3-kinase to insulin/Ras dependent signaling pathways leading to germinal vesicle breakdown and to ascertain whether phosphatidylinositol 3-kinase acts upstream or downstream of Ras in those signaling pathways. We analyzed the process of meiotic maturation induced by progesterone, insulin or micro-injected oncogenic Ras (Lys12) proteins in the presence and absence of specific inhibitors of phosphatidylinositol 3 kinase activity. As expected, the progesterone-induced maturation was independent of phosphatidylinositol 3-kinase since similar rates of germinal vesicle breakdown were produced by the hormone in the presence and absence of wortmannin and LY294002. In contrast, insulin-induced germinal vesicle breakdown was completely blocked by pre-incubation with the inhibitors prior to insulin treatment. Interestingly, similar rates of germinal vesicle breakdown were obtained in Ras (Lys12)-injected oocytes, independently of whether or not they had been pre-treated with phosphatidylinositol 3-kinase inhibitors. The effect of wortmannin or LY294002 on MAPK and Akt activation by progesterone, insulin or Ras was also analyzed. Whereas insulin activated those kinases in a phosphatidylinositol 3-kinase-dependent manner, progesterone and Ras were able to activate those kinases in the absence of phosphatidylinositol 3-kinase activity. Since Ras is a necessary and sufficient downstream component of insulin signaling pathways leading to germinal vesicle breakdown, these observations demonstrate that phosphatidylinositol 3-kinase is not a downstream effector of Ras in insulin/Ras-dependent signaling pathways leading to entry into the M phase in Xenopus oocytes. PMID- 10371207 TI - Identification of new tumor suppressor genes based on in vivo functional inactivation of a candidate gene. AB - As a step towards developing a new functional test for the identification of tumor suppressor genes, human wild type and mutant RB genes were expressed in the mouse A9 fibrosarcoma cell line under the transcriptional regulation of the tetracycline repressor using two new vectors: pLNCtTA and pETI. Following passage of the transfectants in immunodeficient SCID mice, the wild type RB gene was deleted or functionally inactivated already after the first passage in all 20 tumors tested. In contrast, a non-functional mutant RB gene was maintained in all 10 tumors studied. These results suggest that tests for the identification of tumor suppressor genes may be based on their functional inactivation in vivo, rather than on growth suppression. PMID- 10371208 TI - Schizosaccharomyces pombe UDP-galactose transporter: identification of its functional form through cDNA cloning and expression in mammalian cells. AB - The Schizosaccharomyces pombe UDP-galactose transporter cDNA (SpUGT cDNA), encoding the product of the gms1+ gene which consists of two exon sequences separated by a 173-bp intron, was cloned by RT-PCR. Its product, a hydrophobic protein of 353 amino acid residues resembling its human counterpart, was expressed in the Golgi membranes of UDP-galactose transporter-deficient Lec8 cells, and complemented the genetic defect of the mutant cells. This indicated that SpUGT cDNA encodes the functional S. pombe UDP-galactose transporter. The product of an ORF found in the second exon, which was previously assumed to be the S. pombe UDP-galactose transporter, thus represents an inactive, truncated form of the SpUGT protein. PMID- 10371210 TI - An 86 kDa diapause protein 1-like protein is a component of early-staged encapsulation-relating proteins in coleopteran insect, Tenebrio molitor larvae. AB - Recently, we reported two novel early-staged encapsulation-relating proteins (56 kDa and 48 kDa ERPs) isolated from the hemolymph of coleopteran insect, Tenebrio molitor larvae [Cho et al. (1999) Eur. J. Biochem. (in press)]. Here, a cDNA clone for another early-staged encapsulation-relating protein (86 kDa) was isolated. We found that the 86 kDa protein shows high homology with insect diapause protein 1. The 86 kDa protein was localized in the fat body and hemolymph, but not hemocyte lysate. A significant level of 86 kDa protein was detected in pre-pupae stage, but it decreased rapidly at late larvae and pupae, and no protein was found in embryo, early larvae and adult stages. This diapause protein 1-like protein is likely to be a component of early-staged encapsulation relating proteins in the insect cellular defense reaction. PMID- 10371209 TI - Nucleotide sequence of a cDNA encoding a common precursor of disintegrin flavostatin and hemorrhagic factor HR2a from the venom of Trimeresurus flavoviridis. AB - The venom of Trimeresurus flavoviridis has three disintegrins that act as platelet aggregation inhibitors by binding to integrin alphaIIb beta3 on platelets through its Arg-Gly-Asp sequence. We isolated the cDNA encoding the flavostatin precursor that is one of the disintegrins in T. flavoviridis venom. The open reading frame consisted of four regions, a pre-peptide region, a metalloprotease region, a spacer region and a disintegrin region, indicating that the flavostatin precursor belongs to the metalloprotease/disintegrin family. Surprisingly, the deduced amino acid sequence of the metalloprotease region was completely consistent with that of hemorrhagic metalloprotease HR2a, which indicated that this metalloprotease released from the flavostatin precursor functions as a hemorrhagic factor. These observations indicated that a disintegrin and a hemorrhagic metalloprotease were synthesized as a common precursor. Thus, our results support the hypothesis that a disintegrin is synthesized as a metalloprotease/disintegrin precursor and matures by cleavage from the precursor molecule. PMID- 10371212 TI - Morphological and biochemical analysis of the secretory pathway in melanoma cells with distinct metastatic potential. AB - In this report, we have investigated whether alterations of the morphological and functional aspects of the biosecretory membrane system are associated with the metastatic potential of tumor cells. To this end, we have analyzed the morphology of the Golgi complex, the cytoskeleton organization and membrane trafficking steps of the secretory pathway in two human melanoma A375 cell line variants with low (A375-P) and high metastatic (A375-MM) potential. Immunofluorescence analysis showed that in A375-P cells, the Golgi complex showed a collapsed morphology. Conversely, in A375-MM cells, the Golgi complex presented a reticular and extended morphology. At the ultrastructural level, the Golgi complex of A375-P cells was fragmented and cisternae were swollen. When the cytoskeleton was analyzed, the microtubular network appeared normal in both cell variants, whereas actin stress fibers were largely absent in A375-P, but not in A375-MM cells. In addition, the F-actin content in A375-P cells was significantly lower than in A375-MM cells. These morphological differences in A375-P cells were accompanied by acceleration and an increase in the endoplasmic reticulum to Golgi and the trans-Golgi network to cell surface membrane transport, respectively. Our results indicate that in human A375 melanoma cells, metastatic potential correlates with a well-structured morphofunctional organization of the Golgi complex and actin cytoskeleton. PMID- 10371211 TI - Expression analysis of the thyrotropin-releasing hormone receptor (TRHR) in the immune system using agonist anti-TRHR monoclonal antibodies. AB - Monoclonal anti-rat thyrotropin-releasing hormone (TRH) receptor (TRHR)-specific antibodies (mAb) were generated by immunization with synthetic peptides of rat TRHR partial amino acid sequences; one (TRHR01) was directed against a sequence (84-98) in the extracellular portion of the rat TRHR reported to be constant among different species, including man, and the second (TRHR02) recognizes the C terminal region sequence 399-412. In lysates from GH4C1 cells, a clonal rat pituitary cell line, both mAb recognize the TRHR in Western blot analysis, and TRHR02 immunoprecipitates the TRHR. Incubation of GH4C1 cells with the mAb causes a fluorescence shift in fluorescence-activated cell sorting analysis. The cells were stained specifically by both mAb using immunocytochemical techniques. Furthermore, TRHR01 is agonistic in its ability to trigger Ca2+ flux, and desensitizes the TRH receptor. We tested for TRHR in several rat organs and found expression in lymphoid tissues. TRHR01 recognizes the human TRHR, and analysis of human peripheral blood lymphocyte and tonsil-derived leukocyte populations showed receptor expression in non-activated and phytohemagglutinin-activated T and B cells. PMID- 10371213 TI - Identification of cold-sensitive mutations in the Schizosaccharomyces pombe actin locus. AB - In recent years, the actin cytoskeleton in Schizosaccharomyces pombe has become the subject of intense scrutiny. However, to date, only a single actin mutation has been identified. Described here is the isolation and characterization of four new cold-sensitive actin mutations. Sequence analysis of the mutant actin genes indicated that each of these mutations caused alterations in single amino acids that are conserved in all actin sequences. These mutants differ in their phenotypes. One of these mutations (act1-48) was identified as an extragenic suppressor of a mutation in the cdc4 gene, which is required for actin ring formation and cytokinesis. Interestingly, when act1-48 mutant cells were shifted to the restrictive temperature, actin patches were not detected but the actin ring formation and stability was unaffected. The three other mutations, act1-16, act1-32 and act1-67, primarily affected the actin ring formation or stability while F-actin patches did not seem to be substantially different in appearance. Given that the ultrastructural architectures of F-actin patches and the F-actin ring are presently unclear, these mutations, which affect one structure or the other, should be useful for future studies on the role of actin itself in the function of these F-actin-containing structures in S. pombe. PMID- 10371214 TI - Human histamine H2 receptor gene: multiple transcription initiation and tissue specific expression. AB - We have characterized the genomic structure of 12.3 kb of the 5'-flanking region of the human histamine H2 receptor gene. The multiple transcription initiation sites of the human histamine H2 receptor gene were mapped utilizing the 5'-end cap structure of mRNA. We found that a 85 bp segment (-610-(-)525 bp) immediately upstream of the initiation site exhibits a strong promoter activity in the gastric adenocarcinoma, MNK45, expressing human histamine H2 receptor. A 4.8 kb transcript of the human histamine H2 receptor gene was found in the placenta, spinal cord, lymph node and bone marrow in addition to the previously reported tissues including the heart, brain and stomach, whereas a 1.8 kb transcript was observed in almost all tissues examined. 3'-rapid amplification of cDNA ends revealed the corresponding length of the 3'-untranslated region. These results suggest that the 3'-untranslated region may be involved in the differential expression. PMID- 10371215 TI - Caspase-3 is necessary and sufficient for cleavage of protein synthesis eukaryotic initiation factor 4G during apoptosis. AB - Induction of apoptosis BJAB cells is accompanied by the rapid cleavage of protein synthesis eukaryotic initiation factor 4G and the appearance of a fragment of approximately 76 kDa. Inhibition of apoptotic proteases (caspases) has previously been shown to prevent the cleavage of eukaryotic initiation factor 4G. In MCF-7 breast carcinoma cells, which are deficient in caspase-3, eukaryotic initiation factor 4G is not cleaved but in vivo expression of caspase-3 restores eukaryotic initiation factor 4G cleavage following induction of apoptosis. Recombinant caspase-3 can also cleave eukaryotic initiation factor 4G to yield the 76 kDa fragment both in cell extracts and when the eukaryotic initiation factor 4G is presented in a purified eukaryotic initiation factor 4F complex. These results indicate that caspase-3 activity is necessary and sufficient for eukaryotic initiation factor 4G degradation. PMID- 10371216 TI - Activation of mitogen-activated protein kinase by the bradykinin B2 receptor is independent of receptor phosphorylation and phosphorylation-triggered internalization. AB - Recent evidence suggests that serine/threonine phosphorylation and internalization of beta2-adrenergic receptors play critical roles in signalling to the mitogen-activated protein kinase cascade. To investigate whether this represents a general mechanism employed by G protein-coupled receptors, we studied the requirement of these processes in the activation of mitogen-activated protein kinase by G alpha(q)-coupled bradykinin B2 receptors. Mutant B2 receptors impaired in receptor phosphorylation and internalization are fully capable to activate mitogen-activated protein kinase. Bradykinin-induced long-term effects on mitogenic signalling monitored by measuring the transcriptional activity of Elk1 were identical in cells expressing the wild-type or mutant B2 receptors. Therefore, G protein-coupled bradykinin receptors activate the mitogen-activated protein kinase pathway independently of receptor phosphorylation and internalization. PMID- 10371217 TI - Surface-modified mutants of cytochrome P450cam: enzymatic properties and electrochemistry. AB - We report the electrochemistry of genetic variants of the haem monooxygenase cytochrome P450cam. A surface cysteine-free mutant (abbreviated as SCF) was prepared in which the five surface cysteine residues Cys-58, Cys-85, Cys-136, Cys 148 and Cys-334 were changed to alanines. Four single surface cysteine mutants with an additional mutation, R72C, R112C, K344C or R364C, were also prepared. The haem spin-state equilibria, NADH turnover rates and camphor-hydroxylation properties, as well as the electrochemistry of these mutants are reported. The coupling of a redox-active label, N-ferrocenylmaleimide, to the single surface cysteine mutant SCF-K344C, and the electrochemistry of this modified mutant are also described. PMID- 10371218 TI - Hydrogen peroxide generation by higher plant mitochondria oxidizing complex I or complex II substrates. AB - The generation of H2O2 by isolated pea stem mitochondria, oxidizing either malate plus glutamate or succinate, was examined. The level of H2O2 was almost one order of magnitude higher when mitochondria were energized by succinate. The succinate dependent H2O2 formation was abolished by malonate, but unaffected by rotenone. The lack of effect of the latter suggests that pea mitochondria were working with a proton motive force below the threshold value required for reverse electron transfer. The activation by pyruvate of the alternative oxidase was reflected in an inhibition of H2O2 formation. This effect was stronger when pea mitochondria oxidized malate plus glutamate. Succinate-dependent H2O2 formation was ca. four times lower in Arum sp. mitochondria (known to have a high alternative oxidase) than in pea mitochondria. An uncoupler (FCCP) completely prevented succinate dependent H2O2 generation, while it only partially (40-50%) inhibited that linked to malate plus glutamate. ADP plus inorganic phosphate (transition from state 4 to state 3) also inhibited the succinate-dependent H2O2 formation. Conversely, that dependent on malate plus glutamate oxidation was unaffected by low and stimulated by high concentrations of ADP. These results show that the main bulk of H2O2 is formed during substrate oxidation at the level of complex II and that this generation may be prevented by either dissipation of the electrochemical proton gradient (uncoupling and transition state 4-state 3), or preventing its formation (alternative oxidase). Conversely, H2O2 production, dependent on oxidation of complex I substrate, is mainly lowered by the activation of the alternative oxidase. PMID- 10371219 TI - The thermodynamics and kinetics of electron transfer in the cytochrome P450cam enzyme system. AB - In anaerobic environments the first electron transfer in substrate-free P450cam is known to be thermodynamically unfavourable, but in the presence of dioxygen the reduction potential for the reaction shifts positively to make electron transfer thermodynamically favourable. Nevertheless a slower rate of electron transfer is observed in the substrate-free P450cam compared to substrate-bound P450cam. The ferric haem centre in substrate-free P450cam changes from six co ordinate to five co-ordinate when reduced whereas in substrate-bound P450cam the iron centre remains five co-ordinate in both oxidation states. The slower rate of electron transfer in the substrate-free P450cam is therefore attributed to a larger reorganisation energy as predicted by Marcus theory. PMID- 10371220 TI - Reproducibility in genome sequence annotation: the Plasmodium falciparum chromosome 2 case. PMID- 10371221 TI - Comment concerning the hypothesis: Origins of globular structure in proteins by Vladimir Tolstoguzov. PMID- 10371222 TI - Which blood pressure measurement is more important in the elderly? PMID- 10371223 TI - Treatment options for the weight-conscious smoker. PMID- 10371224 TI - Case of the month: the rest of the story. PMID- 10371225 TI - Newer drug therapy for congestive heart failure. AB - BACKGROUND: The management of congestive heart failure has undergone a number of modifications over the past 5 to 10 years. METHODS: These include assaying the role of inotropic drugs, evaluating the role of phosphodiesterase inhibitors, considering the role of intermittent inotropic infusion in ambulatory patients, and recognizing the importance of angiotensin-converting enzyme inhibitors. Very recently, the important role of angiotensin II receptor blocking agents and the use of beta blockade have provided additional modalities for the control of congestive heart failure. The relative usefulness of such therapy has been reviewed in this article. CONCLUSION: The management of congestive heart failure has undergone considerable change with the use of newer and more effective drugs. PMID- 10371226 TI - The challenge of drug-induced aseptic meningitis. AB - Several drugs can induce the development of aseptic meningitis. Drug-induced aseptic meningitis (DIAM) can mimic an infectious process as well as meningitides that are secondary to systemic disorders for which these drugs are used. Thus, DIAM constitutes a diagnostic and patient management challenge. Cases of DIAM were reviewed through a MEDLINE literature search (up to June 1998) to identify possible clinical and laboratory characteristics that would be helpful in distinguishing DIAM from other forms of meningitis or in identifying a specific drug as the culprit of DIAM. Our review showed that nonsteroidal anti inflammatory drugs (NSAIDs), antibiotics, intravenous immunoglobulins, and OKT3 antibodies (monoclonal antibodies against the T3 receptor) are the most frequent cause of DIAM. Resolution occurs several days after drug discontinuation and the clinical and cerebrospinal fluid profile (neutrophilic pleocytosis) do not allow DIAM to be distinguished from infectious meningitis. Nor are there any specific characteristics associated with a specific drug. Systemic lupus erythematosus seems to predispose to NSAID-related meningitis. We conclude that a thorough history on prior drug intake must be conducted in every case of meningitis, with special focus on those aforementioned drugs. If there is a suspicion of DIAM, a third-generation cephalosporin seems a reasonable treatment option until cerebrospinal fluid cultures are available. PMID- 10371227 TI - Profile for estimating risk of heart failure. AB - CONTEXT: We devised a risk appraisal function to assess the hazard of heart failure in persons who are predisposed by coronary disease, hypertension, or valvular heart disease. OBJECTIVE: To provide general practitioners and internists with a cost-effective method to select people at high risk who are likely to have impaired left ventricular systolic function and may therefore require further evaluation and aggressive preventive measures. METHODS: The routinely measured risk factors used in constructing the heart failure profile include age, electrocardiographic left ventricular hypertrophy, cardiomegaly on chest x-ray film, heart rate, systolic blood pressure, vital capacity, diabetes mellitus, evidence of myocardial infarction, and valvular disease or hypertension. Based on 486 heart failure cases during 38 years of follow-up, 4 year probabilities of failure were computed using the pooled logistic regression model for each sex; a simple point score system was employed. A multivariate profile was also produced without the vital capacity or chest x-ray film because these may not be readily available in some clinical settings. RESULTS: Using the risk factors that make up the multivariate risk formulation-derived from ordinary office procedures-the probability of developing heart failure can be estimated and compared with the average risk for persons of the same age and sex. Using this risk profile, 60% of events in men and 73% in women occurred in subjects in the top quintile of multivariate risk. CONCLUSIONS: Using this multivariate risk formulation, it is possible to identify high-risk candidates for heart failure who are likely to have a substantial yield of positive findings when tested for objective evidence of presymptomatic left ventricular dysfunction. The risk profile may also identify candidates who are at high risk for heart failure because of multiple, marginal risk factor abnormalities that might otherwise be overlooked. PMID- 10371228 TI - The long-term prognostic significance of repeated blood pressure measurements in the elderly: SPAA (Studio sulla Pressione Arteriosa nell'Anziano) 10-year follow up. AB - BACKGROUND: In young and middle-aged people, both systolic (SBP) and diastolic (DBP) blood pressure have a continuous, strong, and independent relationship with subsequent cardiovascular morbidity and mortality. These relationships are not well documented in older people and, until now, studies in the elderly do not provide homogeneous results on the importance of DBP compared with SBP as a cardiovascular risk factor. OBJECTIVE: To determine whether SBP and DBP are independent indicators of mortality risk in the elderly. DESIGN: An observational prospective cohort study to analyze the long-term prognostic significance of repeated SBP and DBP measurements in the elderly. PATIENTS AND METHODS: A total of 3858 outpatients 65 years or older (mean age [SD], 72.9 [4.9] years, 43.5% men) were selected randomly by 444 Italian National Health Service general practitioners in 1983. The population was followed up for 10 years. Crude and adjusted incidence rates of total and cardiovascular mortality were analyzed for classes of SBP and DBP based on the values recorded at the 2 initial visits 1 week apart and those measured during the first 12 months of follow-up. RESULTS: During the 10-year follow-up, 74 patients (1.9%) were lost to follow-up and 1561 (41.3%) died, 709 (45.4% of all deaths) from cardiovascular causes. A positive continuous, graded, strong, and independent association was observed with both total (P<.001) and cardiovascular (P<.001) mortality for SBP but not for DBP. The pattern was similar in both sexes, in persons younger and older than 75 years, regardless of preexisting cardiovascular diseases, and whether they had been receiving antihypertensive treatment at baseline. There was no J-shaped mortality curve in the subjects with the lowest SBP and DBP. CONCLUSIONS: These findings suggest that SBP, but not DBP, is a strong, positive, continuous, independent indicator of mortality risk in the elderly and should be stressed much more than DBP in the diagnosis and treatment of hypertension in this age group. PMID- 10371229 TI - Vertebral fractures and mortality in older women: a prospective study. Study of Osteoporotic Fractures Research Group. AB - BACKGROUND: Osteoporotic fractures, including clinically detected vertebral fractures, are associated with increased mortality. However, only one third of vertebral fractures are diagnosed. It is unknown whether vertebral fractures, whether clinically apparent or not, are associated with greater mortality. OBJECTIVES: To test the hypothesis that women with prevalent vertebral fractures have greater mortality than those without fractures and to describe causes of death associated with vertebral fractures. DESIGN: Prospective cohort study with mean follow-up of 8.3 years. SETTING: Four clinical centers in the United States. PARTICIPANTS: A total of 9575 women aged 65 years or older and enrolled in the Study of Osteoporotic Fractures. MEASUREMENTS: Vertebral fractures by radiographic morphometry; calcaneal bone mineral density; demographic, medical history, and lifestyle variables; blood pressure; and anthropometric measures. In a subset of 606 participants, thoracic curvature was measured during a second clinic visit. MAIN OUTCOME MEASURES: Hazard ratios for mortality and cause specific mortality. RESULTS: At baseline, 1915 women (20.0%) were diagnosed as having vertebral fractures. Compared with women who did not have a vertebral fracture, women with 1 or more fractures had a 1.23-fold greater age-adjusted mortality rate (95% confidence interval, 1.10-1.37). Mortality rose with greater numbers of vertebral fractures, from 19 per 1000 woman-years in women with no fractures to 44 per 1000 woman-years in those with 5 or more fractures (P for trend, <.001). In particular, vertebral fractures were related to the risk of subsequent cancer (hazard ratio, 1.4;95% confidence interval, 1.1-1.7) and pulmonary death (hazard ratio, 2.1;95% confidence interval, 1.4-3.0). In the subset of women who underwent thoracic curvature measurements, severe kyphosis was also related to pulmonary deaths (hazard ratio, 2.6;95% confidence interval, 1.3-5.1). CONCLUSION: Women with radiographic evidence of vertebral fractures have an increased mortality rate, particularly from pulmonary disease and cancer. PMID- 10371230 TI - Economic analysis of low-dose heparin vs the low-molecular-weight heparin enoxaparin for prevention of venous thromboembolism after colorectal surgery. AB - BACKGROUND: Low-dose heparin and low-molecular-weight heparin are effective strategies for preventing venous thromboembolism in colorectal surgery. The economic attractiveness of these 2 strategies in North America is unknown. We conducted an economic analysis of low-dose heparin calcium compared with enoxaparin sodium, a low-molecular-weight heparin, for thromboembolism prophylaxis after colorectal surgery. METHODS: We used decision analysis, with an economic perspective of a third-party payer. Efficacy data were obtained from the Canadian Multicentre Colorectal Deep Vein Thrombosis Prophylaxis Trial and a literature review. Canadian costs for diagnosis and treatment of deep vein thrombosis (DVT), pulmonary embolism (PE), and major bleeding were obtained from chart review and a national hospital database of colorectal surgery; American costs were obtained from published literature. The main outcomes were incremental benefits (symptomatic DVTs, symptomatic PEs, and major bleeding events avoided) and incremental costs for every 1000 patients treated. RESULTS: In the Canadian Colorectal Trial, the relative risk of DVT and PE for enoxaparin compared with low-dose heparin was 1.0 (95% confidence interval, 0.7-1.5), and the relative risk of major bleeding was 1.8 (95% confidence interval, 0.8-3.9). With the use of these data in the baseline analysis, a strategy of enoxaparin prophylaxis was associated with equal numbers of symptomatic DVTs and PEs, and an excess of 12 major bleeding episodes for every 1000 patients treated, with an additional cost of $86 050 (Canadian data) or $145 667 (US data). In a sensitivity analysis using optimal assumptions for efficacy and safety of enoxaparin (relative risk of DVT, 0.8; relative risk of PE, 0.4; relative risk of major bleeding, 1.0), a strategy of enoxaparin prophylaxis was associated with 0.8 fewer symptomatic DVT, 3 fewer symptomatic PEs, and equal numbers of major bleeding episodes for every 1000 patients treated, with an additional cost of $15 217 (Canadian data) or $107 614 (US data). CONCLUSION: Although heparin and enoxaparin are equally effective, low dose heparin is a more economically attractive choice for thromboembolism prophylaxis after colorectal surgery. PMID- 10371231 TI - The efficacy of exercise as an aid for smoking cessation in women: a randomized controlled trial. AB - BACKGROUND: Smoking prevalence rates among women are declining at a slower rate than among men. OBJECTIVE: To determine if exercise, a healthful alternative to smoking, enhances the achievement and maintenance of smoking cessation. METHODS: Two hundred eighty-one healthy, sedentary female smokers were randomly assigned to either a cognitive-behavioral smoking cessation program with vigorous exercise (exercise) or to the same program with equal staff contact time (control). Subjects participated in a 12-session, group-based smoking cessation program. Additionally, exercise subjects were required to attend 3 supervised exercise sessions per week and control subjects were required to participate in 3 supervised health education lectures per week. Abstinence from smoking was based on self-report, was verified by saliva cotinine level, and was measured at 1 week after quit day (week 5), end of treatment (week 12), and 3 and 12 months later (20 and 60 weeks after quit day, respectively). RESULTS: Compared with control subjects (n = 147), exercise subjects (n = 134) achieved significantly higher levels of continuous abstinence at the end of treatment (19.4% vs 10.2%, P = .03) and 3 months (16.4% vs 8.2%, P=.03) and 12 months (11.9% vs 5.4%, P=.05) following treatment. Exercise subjects had significantly increased functional capacity (estimated VO2 peak, 25+/-6 to 28+/-6, P<.01) and had gained less weight by the end of treatment (3.05 vs 5.40 kg, P = .03). CONCLUSIONS: Vigorous exercise facilitates short- and longer-term smoking cessation in women when combined with a cognitive-behavioral smoking cessation program. Vigorous exercise improves exercise capacity and delays weight gain following smoking cessation. PMID- 10371232 TI - Sex differences among adults presenting to the emergency department with acute asthma. Multicenter Asthma Research Collaboration Investigators. AB - BACKGROUND: Asthma is an increasing problem worldwide, particularly among women. Sex differences in acute asthma presentation, management, or outcome would have important medical and economic implications. OBJECTIVE: To compare emergency department (ED) visits for acute asthma among women vs men. METHODS: We performed a prospective cohort study as part of the Multicenter Asthma Research Collaboration. Patients in the ED, aged 18 to 54 years, who presented with acute asthma underwent a structured interview in the ED and another by telephone 2 weeks later. The study was performed at 36 EDs in 18 states. Pregnant women with asthma were excluded (n=53). RESULTS: Of 1228 patients, 64.3% were women. Women did not differ significantly from men by age or education level, but women were more likely to be insured, have a primary care provider, and use inhaled corticosteroids. Women had a higher mean+/-SD peak expiratory flow rate than men, both early (expressed as percent predicted) (53%+/-21% vs 41%+/-18%; P<.001) and late (77%+/-24% vs 65%+/-21%; P<.001) in the ED stay. Despite this, women were more likely to be admitted to a hospital (multivariate odds ratio, 2.2; 95% confidence interval; 1.3-4.0) than men. At 2-week follow-up, women had not experienced more relapse events (odds ratio, 1.1) but were 1.5 times more likely to report an ongoing exacerbation (95% confidence interval; 1.0-2.4). CONCLUSIONS: Of adults who presented to the ED with acute asthma, women were almost twice as common as men. Although men received less outpatient care and had worse pulmonary function, women were more likely to be admitted to the hospital and to report an ongoing exacerbation at follow-up. Further studies are needed to better understand the complex relationship between sex and acute asthma. PMID- 10371233 TI - Staphylococcus aureus bacteremia among elderly vs younger adult patients: comparison of clinical features and mortality. AB - BACKGROUND: Previous studies give conflicting results regarding the effect of age on outcomes in Staphylococcus aureus bacteremia (SAB). These studies have been limited by retrospective design or small sample size. METHODS: We conducted a prospective cohort study of 385 patients with SAB aged 18 to 90 years. The setting was a large academic medical center. We observed patients from diagnosis of SAB to discharge or death. Discharged patients were contacted 12 weeks after their first positive culture findings. Data were collected on demographics, comorbid conditions, focus of infection, length of stay, and outcome. Primary outcomes were total mortality and death due to SAB. RESULTS: Comparisons were made between 145 patients, aged 66 to 90 years, and 240 patients, aged 18 to 60 years. Forty-three (29.7%) of the elderly patients and 36 (15%) of the younger patients died. Death directly attributable to SAB occurred in 21 (14.5%) older and 15 (6.3%) younger patients. After adjusting for confounding variables, older patients continued to have higher total mortality (odds ratio, 2.21; 95% confidence interval, 1.32-3.70), and higher mortality from SAB (odds ratio, 2.30; 95% confidence interval, 1.13-4.69). Infection with methicillin-resistant S aureus was associated with higher total mortality in the elderly (odds ratio, 2.59; 95% confidence interval, 1.23-5.43). CONCLUSIONS: Staphylococcus aureus bacteremia among the elderly is associated with high mortality. Both total mortality and mortality directly attributable to SAB are more than twice as likely in older patients. Infection with methicillin-resistant S aureus carries a worse prognosis than infection with methicillin-sensitive S aureus in the elderly. PMID- 10371234 TI - A metaregression analysis of the dose-response effect of aspirin on stroke. AB - BACKGROUND: We evaluated whether the risk of stroke depends on aspirin dose in patients with a previous transient ischemic attack or stroke. METHODS: We conducted a metaregression analysis of stroke by using published randomized, placebo-controlled trials. We analyzed studies of patients who had recently had a transient ischemic attack or stroke (ie, secondary prevention). We abstracted data on the treatment regimen and stroke. To evaluate the dose-response relationship, we conducted a metaregression analysis of study-specific risk ratios by means of weighted linear regression. RESULTS: Eleven randomized, placebo-controlled trials contributed a total of 5228 patients randomized to aspirin only and 4401 patients randomized to placebo only. The slope of the dose response curve was virtually flat across a wide range of aspirin doses from 50 to 1500 mg/d (P = .49 for test of slope not =0). Summarizing across studies, aspirin decreases the risk of stroke by about 15% (risk ratio, 0.85;95% confidence interval, 0.77-0.94). CONCLUSIONS: Aspirin reduces the risk of stroke by approximately 15%, and this effect is uniform across aspirin doses from 50 to 1500 mg/d. The lowest effective aspirin dose has not yet been identified, but it could be lower than 50 mg/d. PMID- 10371235 TI - The term "lymphangitic pulmonary metastases" resurrected. PMID- 10371236 TI - Recommendations for diagnosis, evaluation, and monitoring of patients with Gaucher disease. PMID- 10371237 TI - Dialysis or plasmapheresis for acute renal failure due to Africanized honeybee stings. PMID- 10371238 TI - Acupuncture and nicotine withdrawal. PMID- 10371239 TI - Antidepressants and smoking cessation. PMID- 10371240 TI - Benefits of influenza vaccination for bedridden patients. PMID- 10371241 TI - The emergence of the social and behavioral sciences in dentistry: Lois Cohen as principal architect. AB - The role that Lois played at the NIDR and the IADR as an advocate for biopsychosocial research in dentistry cannot be underestimated and remains one of her most cherished and lasting legacies. First, she has steadfastly maintained her vision for dentistry as a major health discipline that continues to mature toward acceptance of responsibility for every aspect of the impact that oral disease could have on the health and welfare of its patients. Next, she has been an exemplary role model as a rigorous social scientist, simultaneously advocating that such research be interdisciplinary and collaborative while reflecting only the highest standards of excellence for research from the social, biologic, and clinical sciences. Through her administrative leadership skills, she has encouraged such a research mission to be incorporated into the long-range planning of the NIDCR, IADR, FDI, and ADA. Such is the esteem in which she is held that respected social scientists have been attracted to dentistry, persuaded by her vision and drawn by her science. For several decades, thanks to Lois as the primary role model, these scientists have been able to develop their own careers and research interests while bringing cadres of new scientists similarly committed to the broadest and deepest understanding of dental and orofacial growth and development and the prevention and management of dental and orofacial conditions as those processes emerge in peoples around the world. PMID- 10371242 TI - A randomized clinical trial of triazolam in 3- to 5-year-olds. AB - Triazolam has shown promise as a sedative agent for use in pediatric dentistry. However, the efficacy of triazolam has not been previously examined in a placebo controlled study. The present clinical trial used a two-group, randomized, double blind study design to compare the efficacy of oral triazolam with that of a placebo. The primary hypothesis tested was that triazolam would reduce negative behaviors of pediatric dental patients compared with a placebo. A secondary hypothesis was that triazolam would increase the efficiency of dental treatment by reducing the need for time-consuming behavior management by the pediatric dentist. The subjects were 54 3- to 5-year-old children, randomly assigned to the drug and placebo groups. The active drug, 0.03 mg/kg triazolam (Halcion), or lactose placebo was given orally 30 min before dental treatment. Behavior management techniques commonly used in pediatric dentistry were used during dental treatment. A single pediatric dentist provided all of the dental treatment. The procedure included an inferior block anesthesia and careful attention to anesthesia effectiveness. All sessions were video-taped and the tapes coded for child and dentist behaviors by an independent observer. There were no statistically significant differences between the groups with respect to completion of dental treatment. There were no significant differences found in either the total time or the percent of time that the subjects exhibited disruptive movements, verbal or non-verbal distress. The total use of time in the dental chair was slightly higher in the placebo than in the drug group due to more time spent preparing the child. Contrary to preliminary reports in the literature, this investigation found little or no improvement in child behavior when triazolam was used as a sedative compared with a placebo. However, triazolam did shorten the length of dental treatment, primarily by reducing dentist time in preparing the child for the dental procedure (e.g., establishing rapport and shaping behavior). PMID- 10371243 TI - Occlusal disharmonies modulate central catecholaminergic activity in the rat. AB - Occlusal disharmonies have classically been thought to be involved in the etiopathogenesis of bruxism, as have, more recently, alterations in central neurotransmission, particularly dopaminergic neurotransmission. However, the connection between these two factors has still not been established. In this study, we assessed the effects of diverse occlusal disharmonies, maintained for either 1 day or 14 days, on neurochemical indices of dopaminergic and noradrenergic activity in the striatum, frontal cortex, and hypothalamus of the rat. The in vivo activity of tyrosine hydroxylase, determined as the accumulation of 3,4-dihydroxyphenylalanine (DOPA), 30 min after the administration of 3 hydroxybenzylhydrazine, a DOPA decarboxylase inhibitor, and dopamine and noradrenaline contents were quantified by high-performance liquid chromatography with electrochemical detection. The wearing of an acrylic cap on both lower incisors for 1 day induced a significant increase in DOPA accumulation in the regions analyzed, with parallel increases in dopamine levels in the hypothalamus and dopamine and noradrenaline in the frontal cortex. After the cap was maintained for 14 days, DOPA accumulation tended to return to control values, except in the left striatum, thereby causing an imbalance between hemispheres. In contrast, 1 or 14 days after the lower left and the upper right incisors were cut, less pronounced changes in catecholaminergic neurotransmission were found in the brain areas studied. Moreover, the cutting of one lower incisor did not modify either DOPA accumulation or dopamine and noradrenaline contents in the striatum or hypothalamus. These results provide experimental evidence of a modulation of central catecholaminergic neurotransmission by occlusal disharmonies, being dependent on the nature of the incisal alteration and on the time during which it was maintained. PMID- 10371244 TI - Delayed recruitment of immunocompetent cells in denervated rat periodontal ligament following experimental tooth movement. AB - It has previously been shown that the number of mononuclear phagocytic cells in the periodontal ligament (PDL) of orthodontically moved rat molars is significantly increased (p < or = 0.05) at 3, 7, and 14 days compared with the controls. Since these changes coincide with increased density of peptidergic nerve fibers, it was of particular interest to investigate a possible relation between the immunocompetent cells and sensory nerve fibers in the PDL of experimentally moved and denervated rat molars. Twenty-two young animals had the first right mandibular molar moved mesially, 7, 14, and 21 days after ipsilateral inferior alveolar nerve axotomy. The left side served as unoperated control. An immunohistochemical procedure was carried out on alternate, serial, cryostat sections with antibodies against CDllb (macrophages, dendritic cells) and class II major histocompatibility complex (MHC) molecules (RT1B). At 7 and 14 days, the number of CD11b+- and RT1B-expressing cells in the denervated PDL showed no significant difference compared with the contralateral side. However, at 21 days, when periodontal tissue re-innervation is established, the number of the investigated immunocompetent cells in the PDL of the denervated and experimentally moved mandibular molars demonstrated a significant difference compared with the contralateral and control molars (p < or = 0.05). It can be concluded that axotomy of the inferior alveolar nerve delays the recruitment of macrophage-like and class II MHC molecule-expressing cells in the PDL of orthodontically moved rat molars. The results further indicate that sensory nerve fibers interact with immunocompetent cells and participate in their mobilization to locally inflamed tissues. PMID- 10371245 TI - Root development in mice lacking functional tissue non-specific alkaline phosphatase gene: inhibition of acellular cementum formation. AB - Tissue non-specific alkaline phosphatase (TNAP) is richly present in developing teeth including the cells of the periodontal ligament. Here, we investigated tooth and root development in mice lacking the TNAP gene. Heterozygous mutants were obtained from The Jackson Laboratory, Animal Resources (Bar Harbor, ME, USA) and bred. TNAP-deficient mice and their littermates were killed from 6 to 25 days after birth and their molar blocks processed for light and electron microscopy. It was observed that the eruption of the incisors into the oral cavity was delayed for 2 to 3 days. Also, the onset of mineralization of the mantle dentin in the roots of the developing molars was delayed for 2 to 3 days. Yet, dentin and enamel formation in the homozygous mutants showed a more or less normal pattern, with the exception of localized enamel hypoplasias. The most conspicuous finding was the defective formation of acellular cementum along the molar roots. Instead of a continuous layer, the cementum was deposited as very thin and irregularly shaped patches around the bases of the periodontal ligament fibers. Sharpey's fibers were short and poorly developed. In contrast, the development of the alveolar bone, the periodontal ligament, and the cellular cementum was seemingly unaffected. It is concluded that TNAP represents an essential factor in mantle dentin mineralization and in the formation of acellular cementum. PMID- 10371246 TI - Anti-proliferative capsular-like polysaccharide antigen from Actinobacillus actinomycetemcomitans induces apoptotic cell death in mouse osteoblastic MC3T3-E1 cells. AB - Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans) has been implicated in the etiology of localized juvenile periodontitis (LJP), and produces a multiplicity of tissue-damaging products. Among those products, the capsular-like polysaccharide antigen (CPA) from A. actinomycetemcomitans is a potent mediator of bone resorption. In fact, this CPA (serotype b) is known to promote osteoclast-like cell formation via interleukin (IL)-1alpha production in mouse marrow cultures. Although osteoblasts complete bone formation, there are few reports focusing on the effect of CPA in bone-forming activity of osteoblasts in inflammatory disease sites. We hypothesized that CPA plays a mediating role in osteoblastic cells. Therefore, the purpose of this study was to examine the effect of CPA from A. actinomycetemcomitans on the mouse osteoblastic cell line MC3T3-E1 and human osteosarcoma SaOS-2 cells. A. actinomycetemcomitans serotype c resulted in a potent dose-dependent inhibition of cell proliferation of both cell lines. Characterization of the antiproliferative activity in the CPA demonstrated that it was not cytotoxic for MC3T3-E1. A 20-hour incubation with CPA-c resulted in a significant increase in apoptotic cell death in the cells, as evaluated by both cellular DNA fragmentation ELISA and FACS analysis. In contrast to the results obtained with a cytokine mixture (tumor necrosis factor-alpha, IL-1beta, and interferon-gamma), no inducible nitric oxide (NO) synthase gene expression or NO release could be detected in MC3T3-E1 after incubation with CPA-c. Further, both CPA-b and -c caused potent induction of apoptosis-related modifiers, e.g., Fas mRNA, whereas bcl-2 mRNA levels were unchanged. Therefore, this study has shown that CPA from A. actinomycetemcomitans contains a potent antiproliferative polysaccharide whose activity is associated with apoptotic cell death in MC3T3 E1, and that CPA per se is an inducer of apoptosis mediated by the Fas system but not by NO. PMID- 10371247 TI - Secretory immunoglobulin A heavy chain presents Galbeta1-3GalNAc binding structures for Actinomyces naeslundii genospecies 1. AB - Adherence of Actinomyces naeslundii ATCC 12104 to hydroxyapatite beads coated with protein fractions of parotid saliva, obtained by gel filtration on S-200 HR columns, showed GalNAcbeta1-3Galalpha-O-ethyl-inhibitable binding to high molecular-weight proteins (Stromberg et al., 1992). The present study investigates the nature of these high-molecular-weight binding proteins and determines their specific ability to mediate adherence to representative strains of Actinomyces species. Strain ATCC 12104 bound specifically in a lactose inhibitable manner to the heavy chain of secretory immunoglobulin A (S-IgA), contained within a high-molecular-weight parotid protein fraction separated on SDS-PAGE and transferred to a solid membrane support. Lactose-inhibitable binding to the heavy chain of S-IgA from human colostrum was also demonstrated. Peanut agglutinin bound to the heavy chain of parotid and colostrum S-IgAs contained on solid support membranes, confirming the presence of Galbeta1-3GalNAc residues on these molecules. Both salivary and colostrum S-IgA aggregated with strain ATCC 12104 in a GalNAcbeta1-3Galalpha-O-ethyl-inhibitable fashion. Further separation of high-molecular-weight salivary proteins on S-500 HR columns showed GalNAcbeta1 3Galalpha-O-ethyl-inhibitable binding to both mucin- and S-IgA-containing fractions. The presence of S-IgA in salivary pellicles formed in vivo on teeth was demonstrated by Western blot analysis of pellicle extracts with anti-IgA antibodies. Among strains representing A. naeslundii genospecies 1 and 2 and A. odontolyticus, only those of genospecies 1 with a particular adherence profile showed efficient GalNAcbeta1-3Galalpha-O-ethyl-inhibitable binding to S-IgA. Thus, oligosaccharides on S-IgA may promote bacterial aggregation (or adherence) and provide a mechanism by which S-IgA can interact with bacteria without prior immunological challenge. PMID- 10371248 TI - The effects of histatin-derived basic antimicrobial peptides on oral biofilms. AB - Susceptibility of bacteria to antimicrobial agents is strongly reduced by the formation of complex biofilms. We investigated whether synthetic histatin analogs with broad-spectrum antibacterial activity in vitro were also active against these complex mixtures of bacteria, as present in saliva and plaque. In a simplified model system for dental plaque, hydroxyapatite discs were placed in a continuous culture system comprised of Streptococcus mutans, S. sanguis, S. salivarius, Actinomyces naeslundii, Veillonella parvula, Fusobacterium nucleatum, and Prevotella intermedia. Ex situ treatment of the biofilms formed on these discs with 100 microg/mL of peptide dhvar4 significantly reduced facultative anaerobic, total anaerobic, and obligate anaerobic Gram-negative counts with 0.8, 0.5, and 0.5 log units, respectively. Ex vivo treatment of salivary bacteria gave reductions of 0.4, 0.7, and 1.5 log units, respectively. For ex vivo treatment of plaque bacteria, reductions of 0.4, 0.4, and 1.4 log units, respectively, were found. In both saliva and plaque samples, obligate anaerobic Gram-negative bacteria were significantly more susceptible to dhvar4 than facultatively anaerobic or anaerobic bacteria as a whole (p=0.013 and p=0.018, for salivary bacteria, and p=0.021 and p=0.020 for plaque bacteria, respectively). Although the oral bacteria are protected by biofilm formation, the synthetic histatin analog caused a significant reduction of viable counts in a model for oral biofilm as well as in isolated oral biofilms. PMID- 10371249 TI - Migration inhibitory factor related protein-8 (MRP-8) is an autocrine chemotactic factor for periodontal ligament cells. AB - Previous studies have suggested that human periodontal ligament (PDL) cells secrete a chemotactic factor which stimulates motility in an autocrine manner. Here we report the partial amino acid sequence of a purified factor which shows 100% homology with human migration inhibitory factor related protein-8 (MRP-8). In addition, reverse-transcription polymerase chain-reaction (RT-PCR) analysis revealed that mRNA encoding MRP-8 was expressed in cultured human PDL cells. To confirm that MRP-8 is chemotactic for PDL cells, we synthesized 25 mer peptides overlapped by 5 amino acids covering the entire MRP-8 protein and tested them for their chemotactic activities. The data indicated that amino acid residues 21-45 showed chemotactic activity for cultured human PDL cells. The maximum chemotactic response was observed at the concentration of 10(-15) mol/mL for human PDL cells. The chemotactic activity was estimated to be approximately 1000-fold higher than that of platelet-derived growth factor (PDGF), insulin-like growth factors-I and II (IGF-I, -II), and epidermal growth factor (EGF) when compared on a molar basis. Since MRP-8 is reported to be produced mainly by neutrophils and monocytes, the result of the current study may suggest another important role of MRP-8 in human PDL cells. PMID- 10371250 TI - Keratin 19 downregulation by oral squamous cell carcinoma lines increases invasive potential. AB - Squamous cell carcinoma (SCC) of the head and neck is the sixth most frequent cancer worldwide. The survival rate is among the lowest of the major cancers and has not improved significantly in the past two decades. Extensive local invasion and regional lymph node metastasis are, in large part, responsible for the poor clinical outcome of these tumors. Keratin intermediate filaments are the most abundant cytoskeletal proteins in SCCs and regulate the migration of normal and transformed epithelial cells. Previous studies have shown that expression of the 40-kDa keratin K19 is dysregulated in SCCs arising from oral epithelium. Immunohistochemical experiments demonstrated that, while normal epithelium and dysplastic lesions expressed abundant K19 protein, invasive SCCs exhibited a patchy or negative staining pattern. We subsequently determined that K19 expression was consistently downregulated in seven SCC lines compared with normal epithelium. We therefore wanted to determine if K19 downregulation affected the invasive phenotype of these cells. We found that SCC lines which do not express K19 are significantly more invasive in vitro than those which retain expression of this gene. Stable expression of the K19 cDNA in K19 negative cell lines altered cell morphology and intercellular adhesiveness, and significantly decreased the number of cells able to migrate through a reconstituted basement membrane. Reduced invasiveness was not due to decreased metalloproteinase activity in the K19-expressing clones. We conclude that K19 overexpression in oral SCCs decreases their invasive potential by diminishing migratory capability. PMID- 10371251 TI - Changes in Bcl-2 and Bax expression in rat tongue during 4-nitroquinoline 1-oxide induced carcinogenesis. AB - Bax is considered as a main effector of apoptosis. Bax forms homodimers and also heterodimers with Bcl-2. The function of the Bax-Bax dimer in active cell death is antagonized by Bax-Bcl-2 heterodimers. Thus, the ratio of Bcl-2 and Bax should control the susceptibility of cells to those stimuli that induce apoptotic cell death. An increase in apoptotic change has been shown in many carcinomas. In the present study, the changes in Bcl-2 and Bax expression in the tissue during carcinogenic transformation were examined immunohistochemically by means of the 4 nitroquinoline 1-oxide (4NQO)-induced carcinoma model. Animals were divided into 7 groups of 10 rats each, and given 50 ppm 4NQO solution as drinking water for 4, 8, 12, 16, 20, or 24 weeks. Ten animals were used as controls. Gradual increases in the numbers of Bcl-2- and Bax-positive cells were shown corresponding to the progression of experimental carcinogenesis. Statistically significant differences in Bcl-2 and Bax expression were demonstrated between control and four-week treatment groups (p < 0.01), and between control and eight-week treatment groups (p < 0.05), respectively. Levels of both proteins remained high after the period of dysplastic change of the epithelium. In conclusion, Bcl-2 and Bax are involved in the progression of 4NQO-induced carcinoma. PMID- 10371252 TI - Refinement of the dentinogenesis imperfecta type II locus to an interval of less than 2 centiMorgans at chromosome 4q21 and the creation of a yeast artificial chromosome contig of the critical region. AB - Dentinogenesis imperfecta type II is an autosomal-dominant disorder of dentin formation which has been mapped to the 6.6 centiMorgan D4S2691-D4S2692 interval at human chromosome 4q21. In the current investigation, the use of four short tandem repeat polymorphisms has allowed the critical region to be refined to an interval of less than 2 centiMorgans defined by recombination events in unrelated, affected individuals from two families both of which show independent evidence for linkage to chromosome 4q21. The creation of a yeast artificial chromosome contig of this newly defined interval has allowed us to demonstrate that the critical region encompasses approximately 2 Mb of DNA and that the dentin-specific gene, dentin sialoprotein, maps to this interval within 300 kb of dentin matrix acidic phosphoprotein 1 and bone sialoprotein. Moreover, dentin sialoprotein shows no recombination with the dentinogenesis imperfecta type II phenotype. Dentin sialoprotein is therefore a candidate for the dentinogenesis imperfecta type II locus. PMID- 10371253 TI - Genetic linkage of the dentinogenesis imperfecta type III locus to chromosome 4q. AB - Dentinogenesis imperfecta type III (DGI-III) is an autosomal-dominant disorder of dentin formation which appears in a tri-racial southern Maryland population known as the "Brandywine isolate". This disease has suggestive evidence of linkage to the long arm of human chromosome 4 (LOD score of 2.0) in a family presenting with both juvenile periodontitis and DGI-III. The purpose of this study was to screen a family presenting with only DGI-III to determine if this locus was indeed on chromosome 4q. Furthermore, we wanted to determine if DGI-III co-localized with dentinogenesis imperfecta type II (DGI-II), which has been localized to 4q21-q23. Therefore, a large kindred from the Brandywine isolate was identified, oral examination performed, and blood samples collected from 21 family members. DNA from this family was genotyped with 6 highly polymorphic markers that span the DGI-II critical region of chromosome 4q. Analysis of the data yielded a maximum two-point LOD score of 4.87 with a marker for the dentin matrix protein 1 (DMP1) locus, a gene contained in the critical region for DGI-II. Our results demonstrated that the DGI-III locus is on human chromosome 4q21 within a 6.6 cM region that overlaps the DGI-II critical region. These results are consistent with the hypothesis that DGI-II is either an allelic variant of DGI-III or the result of mutations in two tightly linked genes. PMID- 10371254 TI - Recommendations for the use of Lyme disease vaccine. Recommendations of the Advisory Committee on Immunization Practices (ACIP) AB - This report provides recommendations for use of a newly developed recombinant outer-surface protein A (rOspA) Lyme disease vaccine (LYMErix, SmithKline Beecham Pharmaceuticals) for persons aged 15-70 years in the United States. The purpose of these recommendations is to provide health-care providers, public health authorities, and the public with guidance regarding the risk for acquiring Lyme disease and the role of vaccination as an adjunct to preventing Lyme disease. The Advisory Committee on Immunization Practices recommends that decisions regarding vaccine use be made on the basis of assessment of individual risk, taking into account both geographic risk and a person's activities and behaviors relating to tick exposure. PMID- 10371255 TI - Tobacco, alcohol and drug use in eight- to sixteen-year-old twins: the Virginia Twin Study of Adolescent Behavioral Development. AB - OBJECTIVE: This study reports prevalences of lifetime and current alcohol, tobacco and drug use in adolescents; examines associations between substance use and a number of putative risk factors; and estimates the contribution of genetic, shared and unique environmental influences on substance use. METHOD: Substance use data were collected using the Child and Adolescent Psychiatric Assessment on a population sample of 1,412 male and female monozygotic and dizygotic twin pairs, aged 8 through 16, from the Virginia Twin Study of Adolescent Behavioral Development. RESULTS: Heritabilities were estimated to be 84% and 82% for liability to lifetime and current tobacco use, respectively. For alcohol use the role of genes and environment varied according to the context of reporting. Liability to lifetime alcohol use was estimated to be under environmental control, with 71% of the variation shared by members of a twin pair and 29% unique to individual twins. Lifetime alcohol use without the permission of a parent or guardian and current use of alcohol were predominantly explained by genetic factors (h2 = 72% and 74%). The role of genetic factors increased and that of unique environmental factors decreased with increasing severity of alcohol use. Lifetime use of any drug showed a heritability of 45%, with the shared environment accounting for 47% of the variation. Shared environmental factors explained most of the variation in marijuana use. CONCLUSIONS: Genetic factors explained a significant proportion of the variation in the use of tobacco, alcohol and other drugs. Shared environmental factors contributed significantly to lifetime alcohol use and other drug use. PMID- 10371256 TI - The TWEAK: application in a prenatal setting. AB - OBJECTIVE: The TWEAK is a screening instrument used to identify women who are risk drinkers. Potential limitations of previous studies of the TWEAK in the prenatal setting include indirect administration of the instrument to minority, indigent pregnant women. The purpose of this study is to assess the efficacy of the TWEAK when it is given directly to a sample of pregnant women of different socioeconomic backgrounds. METHOD: The original TWEAK, with two different tolerance questions, was administered to a sample of 135 pregnant women enrolled in a study of alcohol use during pregnancy at the obstetrics practices of the Brigham and Women's Hospital in Boston, Massachusetts. RESULTS: The TWEAK, using the first tolerance question (number of drinks before feeling the first effects of alcohol) with the cut point set at more than two drinks, had the best predictive ability for lifetime alcohol diagnoses and risk drinking. The sensitivity of the TWEAK can be increased if the cut point for the first tolerance question is set at two drinks, with some loss of specificity and predictive ability. Medical record assessment was the least sensitive but most specific method of identifying alcohol use by pregnant women. CONCLUSIONS: The TWEAK has promise as a screening instrument for identifying risk drinking during pregnancy. Future work should include testing in other clinical populations. PMID- 10371257 TI - Alcohol availability and homicide in New Orleans: conceptual considerations for small area analysis of the effect of alcohol outlet density. AB - OBJECTIVE: To determine the geographic relation between homicide rate and two competing measures of exposure to alcohol outlets, alcohol outlets per square mile and alcohol outlets per person. METHOD: Homicides occurring in 1994 and 1995 and on-sale and off-sale alcohol outlets with active 1995 licenses were geocoded by address for aggregation at the census tract level. Ecologic analysis of the 155 urban residential census tracts in New Orleans was conducted with controls for potential sociodemographic confounders (% black, % adults unemployed, % unmarried households, and ratio males 15-24/males 35-44). RESULTS: After logarithmic transformation of all study variables, sociodemographic confounders alone accounted for 58% (R2 = .58) of the variance of homicide rates. Adding off sale alcohol outlet density to the models, measured (beta +/- SE) either as outlets per square mile (beta = .211 +/- .062) or outlets per person (beta = .244 +/- .063), yielded strong geographic relations with homicide and increased the amount of variance explained (R2 = .62). A 10% higher off-sale outlet density accounted for a 2.4% higher homicide rate. CONCLUSIONS: Both off-sale alcohol outlets per square mile and off-sale outlets per person demonstrate strong geographic associations with homicide rates among urban residential census tracts in New Orleans. These findings suggest that communities faced with high rates of assaultive violence might consider policy interventions that address alcohol outlet related factors. PMID- 10371258 TI - Domestic violence before and after alcoholism treatment: a two-year longitudinal study. AB - OBJECTIVE: An initial study of 88 male alcoholics and their wives had shown that domestic violence decreased significantly in the year following a behavioral marital therapy (BMT) alcoholism treatment program (see J. Cons. Clin. Psychol. 63: 256-262, 1995). To determine if violence reductions were stable, the present study examined domestic violence during the second year following BMT for the 75 (of the original 88) couples who provided 2-year follow-up data on violence. METHOD: The prevalence and frequency of domestic violence were assessed for 75 male alcoholics and their wives at entry to and at 1 and 2 years after completing BMT. Data on frequency and consequences of alcoholics' drinking were collected for the 2-year follow-up period. Comparison rates of domestic violence for a demographically matched nonalcoholic sample were derived from a nationally representative survey of violence in American families. RESULTS: Husband-to-wife violence occurred in nearly two-thirds of cases in the year before BMT. For both the first and second year after BMT, violence was significantly reduced and the extent of violence was associated with the extent of the alcoholics' drinking. Frequency of posttreatment drinking was positively correlated with violence, and remitted alcoholics no longer had elevated domestic violence levels when compared with matched controls whereas relapsed alcoholics did. Analyses using various assumptions about violence for the 13 cases without violence data showed that sample attrition did not invalidate the present results. CONCLUSIONS: These results indicate that domestic violence decreased after BMT alcoholism treatment. Further, among remitted alcoholics, violence returned to the level experienced by other American families, in the same way that other aspects of marital, family and psychosocial functioning improve after successful treatment of alcoholism. PMID- 10371259 TI - The effects of a cumulative alcohol dosing procedure on laboratory aggression in women and men. AB - OBJECTIVE: This study directly compared the effects of cumulative alcohol dosing procedure on aggression in both women and men. METHOD: Thirteen women and 13 men consumed three beverages 1 hour apart. There were two experimental conditions: (1) a placebo day, when subjects consumed three 240 ml beverages, each containing only 1 ml of alcohol; and (2) an alcohol day, when subjects consumed three 240 ml beverages, each containing 0.35 g/kg of 95% alcohol. Alcohol doses for women were reduced by 8%. Prior to beverage consumption, and periodically after consumption, subjects participated in 25-minute laboratory testing sessions designed to measure aggression. In this paradigm, subjects could earn points by responding on a button, or aggress toward a fictitious opponent who ostensibly subtracted earnings from them. RESULTS: Both women and men showed an increase in aggressive responding after drinking alcohol but not placebo. As a group the greatest increases were observed after consuming the second alcohol drink (BAC = 0.08%). Aggressive responding, however, remained elevated for several hours after alcohol consumption. A post hoc analysis of the data indicated that subjects with high aggression levels under placebo conditions showed the greatest increases in aggression under alcohol conditions. CONCLUSIONS: These results indicate that at least under these conditions, alcohol does increase aggression in both women and men. The aggression-increasing effects of alcohol appear to be long-lasting and specific to individuals with the higher aggressive tendencies while sober. PMID- 10371260 TI - Psychopathy and antisocial personality disorder among alcoholic inpatients. AB - OBJECTIVE: The purpose of this study was to compare the adequacy of two measurement systems--the Revised Psychopathy Checklist (PCL-R) and DSM-III diagnosed antisocial personality disorder (APD)--to distinguish alcoholic inpatients with regard to alcoholism characteristics, criminal activities and psychiatric disorders. METHOD: The 740 patients, who included 440 men, 387 blacks and 199 Hispanics, were admitted to one of five New York State alcohol treatment inpatient centers. Each patient was interviewed, and DSM-III diagnoses and other characteristics were recorded, and trained interviewers completed the PCL-R. RESULTS: There was a statistically nonsignificant association between DSM-III based APD diagnosis and PCL-based psychopathy diagnosis. APD (relative to non APD) alcoholics had an earlier onset of problem drinking, higher levels of pathological drinking and social impairment, and a higher prevalence of familial alcoholism; a similar pattern was not indicated for PCL-R diagnosed psychopaths relative to nonpsychopaths. APD alcoholics also engaged in higher levels of criminal activities and violent acts. APD alcoholics had a higher prevalence of substance abuse disorders, and psychopaths had a higher prevalence of generalized anxiety disorder, panic disorder and schizophrenia. CONCLUSIONS: Distinct subpopulations of alcoholic inpatients are identified via the APD criteria of DSM III and the psychopathy criterion of the PCL-R. The majority of the identified psychopathic alcoholics in this sample were likely to be secondary psychopaths, characterized by features of psychopathy (e.g., callousness, manipulativeness) and emotional dysregulation and/or thought disturbance. PMID- 10371261 TI - The DSM-IV criteria for adolescent alcohol and cannabis use disorders. AB - OBJECTIVE: The aims of this study are to compare DSM-IV criteria for alcohol and cannabis use disorders with its predecessor, DSM-III-R, and to examine the validity of the new criteria in an adolescent drug clinic sample. METHOD: During evaluation, a sample of 772 adolescents (63% boys, 77% white) were administered a structured interview of diagnostic symptoms and additional problem severity measures. Independent staff ratings of problem severity and treatment referral were collected as well. RESULTS: Compared to its predecessor, DSM-III-R, application of the DSM-IV criteria for alcohol and cannabis users resulted in more abuse assignments and fewer dependence assignments. The shift in assignments appeared to be largely due to a lowering of the abuse threshold, rather than to a tightening of the dependence criteria. The external validity data generally supported the DSM-IV abuse and dependence distinction in adolescents, and the newer criteria were as valid as the older criteria. CONCLUSIONS: In contrast to DSM-III-R, the DSM-IV system yields more abuse cases and fewer dependence cases among adolescent alcohol and cannabis abusers. Validity evidence for the new criteria are defensible, yet the findings are seen as a starting point for discussing the need for tailoring substance use disorder criteria for adolescents. PMID- 10371262 TI - Alcohol use and psychosocial well-being among older adults. AB - OBJECTIVE: The objective of this study was to examine the relationships of three dimensions of alcohol consumption (frequency of drinking, usual total weekly consumption and quantity per occasion) with social and leisure status, stress and psychological well-being among people ages 65 and older. METHOD: Nurse interviewers completed in-house surveys with 826 older persons (65% women) in one community (67% response rate of the census sample). RESULTS: Overall, relationships tended to be modest or nil. Those relationships between negative functioning and heavier alcohol use that did emerge tended to be associated with higher quantity drinking (i.e., drinks per drinking day) and to some extent volume, but not with frequency of drinking. Of the psychosocial variables, poorer psychological well-being, especially depression, was most highly correlated with heavier drinking. These results were the same controlling for sex, age, education, health and use of depressant medications. CONCLUSIONS: Higher quantity and volume of alcohol use among older people show a modest positive association with poorer psychological well-being, independent of other potentially confounding variables such as sex, age, health or use of depressant medications. Frequency of drinking, however, was not related to psychosocial status. PMID- 10371263 TI - Adolescent problem drinking: stability of psychosocial and behavioral correlates across a generation. AB - OBJECTIVE: Research conducted in the 1970s demonstrated that Problem Behavior Theory could account for approximately 40% of the variance in problem drinking in both local and national sample studies. The present analyses sought to determine whether the personality, perceived environment, and behavior variables of the framework continue to contribute to the explanation of problem drinking among contemporary American youth. METHOD: Correlational and multiple regression analyses were performed on six separate databases collected at different times between 1972 and 1992. Due to sociodemographic differences among the samples, separate analyses were performed for male and female adolescents, and age, ethnicity and socioeconomic status were statistically controlled. RESULTS: There was considerable consistency across the samples in both the partial correlations and the partial multiple correlations, and this result held for both genders. Not only did the framework account for the same percentage of the variance (40%) in problem drinking in the 1992 data as it did in the 1972 data, but the results for the intervening years were consistent as well. CONCLUSIONS: The consistency of results over a 20-year period confirms that the social-psychological meaning of adolescent involvement in problem drinking has remained stable despite changes in the larger sociohistorical context. PMID- 10371264 TI - Alcohol-specific socialization, parenting behaviors and alcohol use by children. AB - OBJECTIVE: This panel study examined the relations between alcohol-specific socialization by parents (monitoring of alcohol use by children, allowing alcohol use by children at home, communicating against alcohol use and setting rules against alcohol use), general dimensions of parenting behavior (responsiveness and demandingness) and alcohol use by children. METHOD: A sample of 488 fifth grade children reported their perceptions of alcohol-specific socialization by parents, parental responsiveness and parental demandingness. These variables were used to predict alcohol use when children in the panel were in seventh grade. RESULTS: Nineteen percent of seventh-grade children reported alcohol use in the past 30 days. Logistic regression analyses indicated that, after accounting for children's age, sex, single parent status, prior use of alcohol and exposure to parental modeling of alcohol use, the odds of alcohol use were significantly greater among children who perceived no parental monitoring of alcohol use, who had been allowed by parents to have a drink with alcohol at home and who perceived relatively low levels of parental demandingness. Rules against alcohol use, parental communication against alcohol use and parental responsiveness were unrelated to the study outcome. CONCLUSIONS: Parental monitoring of alcohol use by children, family norms regarding alcohol use by children at home and parental ability to set and enforce behavioral rules merit consideration as factors that should be modified by prevention programs. There is a need, however, for additional research that further examines the relations between exposure to such parenting behaviors during childhood and alcohol use during adolescence. PMID- 10371265 TI - Wives' convergence with their husbands' alcohol use: social conditions as mediators. AB - OBJECTIVE: This study examines, in a general population, how the association of wives' alcohol use with their husbands' is mediated by social conditions such as working and child-rearing roles, age, marital happiness and socioeconomic levels. METHOD: Data come from the Quebec Health and Social Survey 1992-93. The sample comprised 6,582 couples; 3,872 couples after weighting. Regression analyses assessed the contribution of husbands' drinking to wives' drinking, independently, as well as in interaction with social conditions. Frequency of drinking in general and frequency of five or more drinks per occasion (5+) were analyzed. RESULTS: Wives' drinking is positively related to their husbands' drinking, both in terms of frequency of drinking and, to a lesser degree, of frequency of 5+. The drinking frequency association is not modified by working or child-rearing roles, nor by age, but is more marked for couples of higher socioeconomic levels and for wives happy with marital life. The frequency of 5+ association is not modified by marital happiness or by wives' working roles, but is more marked for couples of higher socioeconomic levels, for couples with a child at home and for younger women. CONCLUSIONS: This study highlights the contribution of social factors to the association of wives' drinking with their husbands' and suggests that these social factors operate through structuring marital drinking interactions. Further research is needed to clarify the complex social processes that mediate the spousal drinking association. PMID- 10371266 TI - Changes in alcohol consumption patterns following the introduction of credit cards in Ontario liquor stores. AB - OBJECTIVE: In 1994, regulatory changes were introduced in Ontario, Canada, permitting the purchase of alcoholic beverages with credit cards at government operated liquor stores. Two objectives of this study were: (1) to compare the characteristics of credit card shoppers with non credit card shoppers at liquor stores, and (2) to assess whether changes occurred in alcohol consumption patterns among shoppers following the introduction of credit cards. METHOD: Random digit dialing was used to interview 2,039 telephone participants prior to the introduction of credit cards (Time 1); 1,401 of these subjects were contacted 1 year later (Time 2). Independent sample t tests were used to compare credit card shoppers with shoppers not using credit cards, and paired t tests were performed to assess whether drinking behaviors changed from Time 1 to Time 2. RESULTS: The credit card shoppers were more likely than the non credit card shoppers to be highly educated (p < .001) and to have high incomes (p < .05). Credit card shoppers drank an average of 6.3 drinks over the previous week compared with 4.0 drinks among non credit card shoppers (p < .01). Although the overall amount of alcohol consumed among credit card shoppers dropped from 6.7 drinks at Time 1 to 6.3 at Time 2 (NS), credit card shoppers reported drinking significantly more often after credit cards were introduced (p < .05). CONCLUSIONS: The results suggest that credit cards may not present public health problems since significant increases in alcohol consumption among credit card shoppers were not found. PMID- 10371268 TI - A frequency and content analysis of alcohol advertising on Brazilian television. AB - OBJECTIVE: Two studies were conducted with the objective of analyzing the frequency and content of alcoholic beverage advertising on Brazilian television. METHOD: Study 1 presents a frequency analysis based on 84 hours of TV monitoring in which 1,640 commercials and 243 vignettes were recorded between 8:00 PM and 11:00 PM on the two main stations. Study 2 presents a content analysis of 139 alcoholic and 51 nonalcoholic beverage commercials aired in 1992-93, as evaluated by three trained judges. RESULTS: Study 1 showed the relative frequency of alcoholic beverage commercials (4.6%) to be higher than the frequency of other products such as cigarettes, nonalcoholic beverages and medicines, and lower than that of foods and various other items. Frequency of alcoholic "vignettes" (26.6%) was higher than the frequency of all the other product categories. Frequency data were closely matched by marketing investment data for the period. In Study 2, the most frequent themes and appeals present in alcohol commercials were relaxation, national symbolism, conformity, camaraderie and humor. Human models were present in most ads. Product-related themes such as information, quality or tradition were virtually absent, as were messages to drink moderately. However, 7.2% of the alcohol commercials displayed appeals promoting abusive drinking. CONCLUSIONS: The results seem to reflect the minimal regulation of alcohol advertising in Brazil, and a joint effort by health planners, educators, legislators, alcohol industries and advertising agencies is recommended as a necessary step to reduce alcoholic beverage problems in Brazilian society. PMID- 10371267 TI - Alcohol availability and workplace drinking: mixed method analyses. AB - OBJECTIVE: This article investigates the relationship between subjective social and physical availability of alcohol at work and work-related drinking. METHOD: We integrated survey and ethnographic methods to determine if and why physical and social availability of alcohol predicted work-related drinking in a manufacturing plant with approximately 6,000 employees. Survey data were obtained from in-home interviews with 984 randomly selected workers. Respondents were asked about their overall and work-related drinking, their perceptions of the ease of obtaining or consuming alcohol in the plant, the work-related drinking of others and their approval/disapproval of work-related drinking by co-workers. Ethnographic data were obtained from 3 years of periodic onsite observations and semistructured interviews with key informants to investigate factors underlying alcohol availability and drinking at work. RESULTS: Structural equations modeling of the survey data revealed that subjective social availability of alcohol at work, and particularly perceived drinking by friends and co-workers, was the strongest predictor of work-related drinking. Typical frequency and quantity of alcohol consumption and heavy drinking were predictive also. Subjective physical availability of alcohol was not significantly related to drinking at or before work. Findings from the ethnographic analyses explained survey findings and described characteristics of the work culture that served to encourage and support alcohol availability and drinking. CONCLUSIONS: These results are the first to show significant relationships between alcohol availability and drinking at work, to explain dynamics of that relationship and to demonstrate the potential risks of using only quantitative or only qualitative findings as the basis for prevention. PMID- 10371269 TI - The drunkest drinking driver in Sweden: blood alcohol concentration 0.545% w/v. AB - OBJECTIVE: This article examined drinking drivers (N = 81) who had unusually high blood alcohol concentrations (BAC > or = 0.400% w/v) when apprehended. These drinking drivers were investigated in relation to age, gender, weekday and time of day of arrest, amount of alcohol in the body and the rate of disappearance of alcohol from the blood. METHOD: The concentration of ethanol in whole blood was determined in triplicate by headspace gas chromatography and the rate of disappearance of ethanol (burn-off rate) was calculated when two blood specimens were taken about 1 hour apart (mean = 65 minutes, range = 33-110 minutes). RESULTS: Seven of the 81 drunk drivers were women (9%) with a mean age of 40 years (range = 29-52 years) and 74 were men (91%) with a mean age of 44 years (range = 28-68 years). The mean, median and range of BACs for the women were 0.422% w/v, 0.413% w/v and 0.403-0.451% w/v, compared with BACs of 0.425% w/v, 0.416% w/v and 0.400-0.545% w/v for the men, respectively (p > .05). The BAC was not dependent on subjects' age (F = 1.04, 7/73 df, p > .05) nor the weekday when they were apprehended (F = 1.62, 6/74 df, p > .05). Most drunk drivers (49%) were apprehended between 12 noon and 6 PM, 40% were involved in traffic accidents and 17% did not hold a valid driving license. The mean (+/-SD) disappearance rate of ethanol from blood was 0.023% +/- 0.01% w/v per hour (N = 26), with a range of 0.013 to 0.061% w/v per hour. CONCLUSIONS: Drinking alcohol to reach a BAC of 0.400% w/v or more and attempting to drive a motor vehicle indicates an exceptionally high cellular tolerance to the impairment caused by this drug. The alcohol burn-off rate was relatively high in these heavy drinkers (mean = 0.023% w/v per hour), which probably reflects the development of metabolic tolerance as well. PMID- 10371270 TI - Factors associated with planned avoidance of alcohol-impaired driving in high risk men. AB - OBJECTIVE: This study examines the factors associated with planning to avoid alcohol-impaired driving and successful avoidance in high-risk young men. METHOD: A targeted telephone survey was conducted with male drivers aged 21-35 years who consume alcohol and live in areas of the country where alcohol-related traffic fatalities occur frequently (N = 750). Heavy episodic drinking drivers (i.e., report driving after consuming five or more drinks) were oversampled (N = 230). Respondents were surveyed to assess their attitudes, behavior and social support regarding drinking-driving. RESULTS: Multiple logistic regression revealed that men who believe they can consume six drinks or more before it is too dangerous for them to drive were 45% less likely to report planning to avoid drinking driving. Men who believe they can drive safely after heavy episodic drinking were 61% less likely to be successful in avoiding drinking-driving. Having friends who disapprove of driving after heavy episodic drinking and believing a close friend would be successful in preventing drinking-driving were significantly associated with making plans to avoid drinking-driving, although this association was less strong for successful avoidance. Men who had their wife/girlfriend along when they were out drinking were two and one-half times more likely to make plans to avoid drinking-driving. The presence of a wife or girlfriend was an even stronger predictor (multivariate odds ratio = 3.74) of successful avoidance. CONCLUSIONS: Attitude and social factors are associated with drinking-driving avoidance in a group of young men at risk for alcohol-related driving fatalities. Friends and wives/girlfriends appear to influence drinking-driving avoidance behavior in high risk drinking drivers. PMID- 10371271 TI - Muscle performance in detoxified alcoholics. AB - OBJECTIVE: Although numerous reports have described alcohol muscle dysfunction (myopathy) in histochemical and morphological terms, a description of the relative impact of alcoholism on the functional capabilities (strength, power) of the various muscle groups of the arms and legs has not been reported. METHOD: The strength of flexor and extensor muscles of the elbow, knee and ankle was evaluated in a group of healthy detoxified white, alcoholic men (N = 83) and lifestyle control subjects (N = 61) using a computer-operated isokinetic dynamometer. RESULTS: The muscle mass (muscle-plus-bone cross-sectional area) did not differ significantly in alcoholics and controls for any of the limbs studied. Tests of isokinetic torque, work and power revealed small deficits (3-8%) in alcoholics in muscle groups involved in elbow flexion/extension, knee extension and ankle dorsiflexion. Tests of isometric torque in the knee extensors at different degrees of muscle stretch revealed approximately 4-6% lower torque in alcoholics at four different muscle lengths. The force-velocity (isoinertia knee extension) measures indicated slightly slower speeds of contraction (3.9-6.6%) in alcoholics under varying load conditions. CONCLUSIONS: The small magnitude of the muscle performance deficit in this group of alcoholics was less than expected and is probably related to (1) the elimination of subjects with other medical comorbidities or polydrug abuse, (2) the comparison with a control group of somewhat similar lifestyle and (3) the long (35-day) detoxification period elapsing before testing. PMID- 10371272 TI - Suicidal ideation among the United States drinking population: results from the National Longitudinal Alcohol Epidemiologic Survey. AB - OBJECTIVE: Data from a national representative sample of adults was used to identify major risk factors of suicidal ideation among the U.S. drinking population. METHOD: Data from a sample of 18,352 current drinkers, 18 years of age and older, were analyzed by means of multiple logistic regression analysis. In these analyses, multivariate associations were examined between risk factors for suicidal ideation and the occurrence of suicidal ideation. RESULTS: For men and women, past year major depression and alcohol dependence were identified as risk factors of suicidal ideation, with major depression having the more sizable impact. Suicidal ideation was increased among men with a past alcohol use disorder, and elevated among women who had used drugs nonmedically and developed a drug use disorder during the past year. The occurrence of a recent physical illness and lifetime treatment for major depression among men and women increased the risk of suicidal ideation, while marriage was protective against ideation for both sexes. Unemployment and having a family history of alcoholism increased the risk of suicidal ideation among men, but not women. CONCLUSIONS: Major findings are discussed in terms of the impact of severity versus chronicity of psychopathology on suicidal ideation, gender roles and differential engagement in suicidal ideation, and the recognition and treatment of major depression as the single most important intervention in reducing suicidal behavior. PMID- 10371273 TI - Research on complementary medicine in rheumatic diseases: the need for better quality studies and reproduction of claimed positive results. PMID- 10371274 TI - 5'-Nucleotidase as a marker of both general and local inflammation in rheumatoid arthritis patients. AB - OBJECTIVES: To evaluate measurements of serum and synovial fluid 5'-nucleotidase (5'N) activity as a marker of general and local inflammation in arthritis, and to resolve a contradiction in the literature as to whether or not the activity of 5'N in the synovial fluids of rheumatoid arthritis (RA) patients is raised in comparison with that in the synovial fluids of other arthritis patients. METHODS: Assays for 5'N were carried out in the presence of inhibitors of other phosphatases, AMP deaminase and of 5'N itself. RESULTS: The 5'N activity in the synovial fluid of RA patients was both significantly higher (mean 1.7-fold) and had a greater variance than that in the synovial fluids of other arthritis patients, and the contradiction in the literature was resolved. There was a strong correlation between the 5'N activity in the sera of RA patients and their erythrocyte sedimentation rate. There was no significant correlation between the 5'N in the serum and synovial fluid for the RA patients, in marked contrast to the strong correlation between the two 5'N activities shown by the osteoarthritis patients. The 5'N activity was greater in the synovial fluid than in the serum for virtually all the patients, showing that it was being made locally. CONCLUSIONS: The 5'N activity in the serum (which came mostly from the liver) could be used as a marker of general inflammation, whereas the 5'N in the synovial fluid was mostly produced locally, and could be used as a marker of joint inflammation, particularly for the RA patients. PMID- 10371275 TI - Clinical and laboratory manifestations of systemic sclerosis (scleroderma) in Black South Africans. AB - A retrospective study of systemic sclerosis (SSc) in Blacks attending a tertiary hospital on the Witwatersrand, South Africa, was undertaken. The female:male ratio of the 63 patients was 4.6:1 and the mean age of onset of SSc was 36.1 yr. Four of the 11 males were ex-goldminers and nine females resided close to goldmines. Forty-one patients had diffuse cutaneous SSc (dcSSc), 18 had limited cutaneous SSc (lcSSc) and four were unclassified. Overall, 56% had pulmonary fibrosis, 37% had myositis and 98% were antinuclear antibody (ANA) positive, with a notable absence of anti-centromere antibodies. Subset comparisons showed myositis and a reduced forced vital capacity to be significantly more common with dcSSc than lcSSc. The only significant sex differences were that arthralgia/arthritis was more common in women, while calcinosis occurred more frequently in men. Seven of the eight known deaths occurred in patients with dcSSc. These findings, particularly the age of disease onset, predominance of the dcSSc subset, inflammatory features of myositis and a raised erythrocyte sedimentation rate, and absence of anti-centromere antibodies, are similar to those reported previously in African-Americans. PMID- 10371276 TI - Effects of soluble interleukin-1 type II receptor on rabbit antigen-induced arthritis: clinical, biochemical and histological assessment. AB - OBJECTIVES: To investigate the effects of soluble interleukin-1 (IL-1) type II receptor (sIL-1RII) on a number of clinical, biochemical and histological parameters in rabbit antigen-induced arthritis. METHODS: Arthritis was induced by intra-articular injection of methylated bovine serum albumin (mBSA) into rabbits pre-sensitized to the same antigen. An initial i.v. bolus of sIL-1RII was administered, followed by s.c. mini-pump dosing for 14 days, starting at the time of the arthritis induction. Animals received vehicle (saline 500 microl + 5 microl/h), low-dose sIL-1RII (13.4 microg + 1.34 microg/h) or high-dose sIL-1RII (40.2 microg + 4.02 microg/h). RESULTS: Marked, dose-related inhibition of joint diameter, plasma prostaglandin E2 (PGE2), and synovial fluid IL-1alpha and IL 1beta concentrations were seen after administration of sIL-1RII. However, synovial fluid PGE2 concentrations and synovial fluid cell counts were not affected. A significant inhibitory effect was also seen histologically on soft tissue swelling and joint damage with high-dose sIL-1RII. CONCLUSIONS: These results demonstrate that IL-1 plays an important role in the pathogenesis of rabbit antigen-induced arthritis, thus confirming it as an excellent animal model with respect to evaluating anti-cytokine therapies for rheumatoid arthritis. PMID- 10371277 TI - Serum MMP-3 in rheumatoid arthritis: correlation with systemic inflammation but not with erosive status. AB - OBJECTIVE: Metalloproteinases (MMP) play an important role in the remodelling of the extracellular matrix. However, evidence that they are responsible for tissue damage in pathological situations remains circumstantial. Stromelysin (MMP-3) production is increased in rheumatoid arthritis (RA), and has been proposed as a marker of joint damage. The relevance of serum levels of MMP-3 to erosions in RA was studied. METHODS: Fifty-three patients with active RA of > 5 yr duration and with available X-rays were stratified according to disease duration. Hand X-rays were scored for erosions. Patients were then classified into upper and lower quartiles. Serum MMP-3 levels were compared between these two groups. RESULTS: No significant differences in serum MMP-3 were seen between high and low eroders. A statistically significant correlation was observed between sMMP-3 and erthyrocyte sedimentation rate and C-reactive protein. CONCLUSIONS: Serum MMP-3 is not an independent marker of joint damage, but is correlated with systemic inflammation. Its precise role in joint damage in RA remains to be elucidated. PMID- 10371278 TI - Chlamydia pneumoniae as a triggering infection in reactive arthritis. AB - OBJECTIVE: To determine the role of Chlamydia pneumoniae as a triggering infection in reactive arthritis (ReA). METHODS: Sixty patients with acute arthritis were screened for the evidence of triggering infections. In all patients, bacterial stool cultures, culture of Chlamydia trachomatis in urethra/cervix, and/or bacterial serology were studied. Chlamydia pneumoniae antibodies were measured by specific microimmunofluorescence test. RESULTS: Thirty-five of 60 patients fulfilled the diagnostic criteria for ReA. Thirty-one patients had microbial/serological evidence of preceding infection due to Salmonella, Yersinia, Campylobacter or Chlamydia trachomatis, or they had enteritis or urethritis prior to arthritis. Four additional patients had high antibody titre for C. pneumoniae. Three of these four patients had preceding lower respiratory symptoms, and were positive for HLA-B27. The clinical picture of C. pneumoniae-positive ReA patients was similar to that of ReA patients with other definite aetiology. CONCLUSION: Chlamydia pneumoniae is a triggering factor in approximately 10% of patients with acute ReA. PMID- 10371280 TI - Socio-economic consequences of rheumatoid arthritis in the first years of the disease. AB - OBJECTIVE: Few data have been presented to document the impact of rheumatoid arthritis (RA) on socio-economic well-being. In this study, exact figures on socio-economic consequences were assessed. METHODS: The socio-economic consequences were studied in an inception cohort (186 early RA patients, mean disease duration 3 yr) by measuring the change in work capability, income, rest during the daytime, leisure time activity, transport mobility, housing and social support occurring in the first years of the disease. RESULTS: For 89% of the patients, RA had an impact on one of the socio-economic items; for 58%, at least three of these items were affected simultaneously. Work disability appeared to be 4-15 times higher than in the general population. After 3 yr, 42% of the patients were registered as work disabled. Nearly a quarter of the patients experienced income reduction. Over 40% of the patients claimed extra rest during the daytime. Leisure activity changed towards activities with a lower joint load. There was a decline in transport mobility for 52% of the patients. Social support increased strongly. CONCLUSIONS: Socio-economic change already presents in the first years of RA and appears to be influenced by age, gender, marital status and work disability. Furthermore, physical limitation appeared to be predictive for work related income reduction, reduced transport mobility and development of social dependency. PMID- 10371279 TI - Familial aggregation of rheumatoid arthritis in The Netherlands: a cross sectional hospital-based survey. European Consortium on Rheumatoid Arthritis families (ECRAF). AB - OBJECTIVES: To study the familial aggregation of rheumatoid arthritis (RA) in The Netherlands and to analyse the effect of proband characteristics on the concordance rates for RA. Secondary aims were to compare the characteristics of patients in an early RA inception cohort with those of regular patients and to select Dutch families for the genome-wide scan carried out by the European Consortium on RA families (ECRAF). METHODS: A cross-sectional, hospital-based survey aimed to identify affected sibpair (ASP) families among our whole RA population. Familial RA, or an ASP family, was defined by the presence of at least two siblings fulfilling the 1987 ACR criteria for RA. RESULTS: The estimated prevalence for familial RA was 9.8% and similar to that found in previous hospital series. The true-positive reporting rate for RA in sibs was 60%. Sibship size in ASP families (mean +/- S.D. = 7.8+/-3.3) was significantly larger than in the Dutch population. Probands with familial RA were more often rheumatoid factor positive and had a longer follow-up. Male gender and history of joint replacements were associated with higher concordance rates for RA. However, regression analysis showed that, correcting for sibship size, the concordance rate for RA was largely not explained by proband characteristics. Compared to regular RA patients, our inception cohort encompassed more male and/or rheumatoid factor-negative patients, but had similar performance in the study and rate of familial aggregation. CONCLUSIONS: The familial aggregation of RA in The Netherlands is not increased and occurs preferentially in large sibships. Among proband characteristics, sibship size is most clearly related to the recurrence of RA in particular families. Patients' recognition of RA manifestations in relatives is not optimal. PMID- 10371281 TI - A comparative quantitative morphometric study of cell apoptosis in synovial membranes in psoriatic, reactive and rheumatoid arthritis. AB - OBJECTIVES: Inflammatory arthritides/synovitides such as psoriatic (PsA), reactive (ReA) and rheumatoid (RA) arthritis share numerous immunopathological features, but develop different patterns of joint involvement. To investigate whether distinctive cell apoptosis may play a role in this context, we have assessed synovial cell apoptosis in situ in PsA and ReA, and compared it with RA and 'non-inflammatory' controls. METHODS: TdT-mediated dUTP nick end-labelling (TUNEL) of DNA breaks complemented immunoperoxidase staining for CD68 or LCA as the specific cell markers. RESULTS: The proportion of apoptotic synovial lining cells was high in PsA, ReA and RA compared to values in controls (P < 0.05). No differences existed between these inflammatory arthritides in numbers or type of apoptotic lining cells. In RA, however, in contrast to PsA and ReA, apoptotic lining cells were clustered or, in a small subset of samples, were very low in number. Prominent apoptosis of inflammatory cells in the sublining in ReA has accounted for higher overall apoptotic cell numbers in synovial stroma (sublining + perivascular inflammatory cell infiltrates) in this condition than in RA or PsA (P < 0.05). CONCLUSIONS: No disease-specific pattern in the phenotype of apoptotic synovial lining cells could be suggested in any of the inflammatory arthritides studied. However, topological differences in the lining and quantitative differences in the inflammatory cell apoptosis in synovial stroma may in part explain the occurrence of the prominent synovial lining cell hyperplasia distinguishing RA from ReA and PsA. On the other hand, relatively frequent inflammatory cell apoptosis may contribute both to the downregulation of synovial inflammation and to the control of synovial lining hyperplasia in ReA. PMID- 10371282 TI - Biomechanical analysis of posture in patients with spinal kyphosis due to ankylosing spondylitis: a pilot study. AB - OBJECTIVES: Patients with ankylosing spondylitis may experience a progressive spinal kyphosis, which induces a forward and downward displacement of the centre of mass (COM) of the trunk. In this pilot study, the possible mechanisms used to compensate for the displacement of the trunk COM were analysed. METHODS: Joint angles of hip, knee and ankle were determined in four patients with ankylosing spondylitis and compared to data of 18 healthy subjects. Each patient stood on a force platform and had to adopt several predefined postures, which were recorded by a video camera. RESULTS: In three patients, the hips were flexed when standing relaxed, and in all patients hip extension was limited. The knee angles of three patients were smaller and in two patients the angle of the ankles was larger compared to healthy subjects. CONCLUSIONS: The results suggest that the hip joints are at least no longer involved in balance control. This may imply that conservative therapy should focus on the prevention of restriction of the hip joints. PMID- 10371283 TI - New arguments for a vasculitic nature of polymyalgia rheumatica using positron emission tomography. AB - OBJECTIVE: To study the possible contribution of fluorodeoxyglucose (FDG) positron emission tomography (PET) in the diagnosis of giant cell arteritis and polymyalgia rheumatica. METHODS: A consecutive case series consisting of five patients with polymyalgia rheumatica, six patients with temporal arteritis and 23 age-matched patients with other inflammatory conditions were evaluated with FDG PET. Studies were performed before therapy with steroids was started. RESULTS: A total of 4/6 patients with giant cell arteritis and 4/5 patients with polymyalgia had increased FDG uptake in their thoracic vessels, compared to 1/23 controls (P < 0.001). Increased vascular FDG uptake in the upper legs was seen in 8/11 patients with giant cell arteritis or polymyalgia compared to 8/23 control patients (P < 0.05), and in the lower legs in 6/11 patients compared to 6/23 controls (P = not significant). CONCLUSIONS: FDG-PET scan is the first non invasive technique which may indicate large-vessel vasculitis and which can show its extension throughout the body. It strongly suggests that polymyalgia rheumatica is a form of vasculitis. PMID- 10371284 TI - Positive association of the HLA DMB1*0101-0101 genotype with rheumatoid arthritis. AB - OBJECTIVE: HLA DM is a non-classical major histocompatibility complex (MHC) class II molecule that has been shown to facilitate peptide loading with classical class II molecules. METHODS: In this study, we analysed the polymorphism in exon 3 of HLA DMA and DMB genes by a polymerase chain reaction-sequence-specific oligonucleotide probe method in 163 rheumatoid arthritis (RA) patients and 146 ethnically matched controls. The HLA-DRB1 genotype was also analysed by a reverse dot blot method. RESULTS: Our results show in RA patients a significant increase in the HLA DMB*0101 allele frequency (83% vs 72.3% of the controls, P < 1.6 x 10( 3), significance at P < 0.0125) and in the HLA DMB*0101-0101 homozygote genotype frequency [70.8% vs 50% of the controls, P < 4.2 x 10(-4), significance at P < 0.00625, odds ratio (OR) = 2.4, 95% confidence interval (CI): 1.43-4]. The increase in DMB*0101 allele and homozygote genotype frequencies was independent of a linkage disequilibrium between DMB and DRB1 alleles. The analysis of non random associations between the HLA-DM and DRB1 alleles only revealed a significant association in controls between DMB*0104 and DRB1*07 alleles (delta = 0.01, P < 7 x 10(-4), significance at P < 9.6 x 10(-4)). On the other hand, the DMB*0101-0102 genotype frequency was increased in DRB1*0401-negative RA patients as compared to controls (11% vs 2%, P < 0.011, significance at P < 0.015, OR = 6.2, 95% CI: 1.2-30). CONCLUSION: Our data suggest that HLA-DM alleles could play a role in the genetic susceptibility to RA. PMID- 10371285 TI - Porphyria cutanea tarda affecting a rheumatoid arthritis patient treated with methotrexate: association or coincidence? AB - We describe the case of a 44-yr-old woman, suffering from rheumatoid arthritis for 15 yr, who developed porphyria cutanea tarda while being treated with methotrexate. The cutaneous lesions healed and the metabolic anomalies improved after a few months, despite continuing the treatment. PMID- 10371286 TI - Childhood Behcet's disease: clinical features and comparison with adult-onset disease. AB - OBJECTIVE: To study the clinical spectrum of Behcet's disease (BD) in childhood, in comparison to adult-onset disease. METHODS: Nineteen children, who fulfilled disease criteria up to the age of 16 yr, were studied. The results were compared to those of 34 adult patients with BD. An activity index and severity score were calculated for both study groups. RESULTS: The mean age of disease onset was 6.9+/-3.9 yr, similar ages of onset were found in males and females. The clinical spectrum of childhood BD resembled that of adult disease; however, the prevalence of certain manifestations was different between children and adults. Children with BD had significantly less genital ulcers, less vascular thromboses and more non-specific gastrointestinal symptoms, as well as central nervous system involvement and arthralgia. A relatively high prevalence of uveitis was found in childhood BD. The activity index and severity score were significantly lower in children than in adults. CONCLUSION: Our results point to a similar systemic expression of BD in children and adults; however, the disease seems to run a less severe course in children. PMID- 10371287 TI - SAPHO syndrome or psoriatic arthritis? A familial case study. AB - OBJECTIVE: To discuss the relationships between SAPHO (synovitis, acne, pustulosis, hyperostosis and osteitis) syndrome and the group of spondylarthropathies. METHODS: Few reports of familial SAPHO have been published. We describe three children, two sisters and one brother, whose clinical and radiological presentation was in accordance with SAPHO syndrome. RESULTS: Two children developed psoriasis, and one child palmoplantar pustulosis. Both sacroiliac and sternoclavicular joints were involved in these three cases. Some features in our observations are also common to psoriatic arthritis. No association was found with HLA antigens, but a history of trauma preceding the onset of symptoms was present in all three children. CONCLUSIONS: We can consider that SAPHO is nosologically related to spondylarthropathies. Psoriatic arthritis could be the missing link between SAPHO and spondylarthropathies. It is likely that both genetic and environmental factors are involved. PMID- 10371288 TI - Evaluation of tumour necrosis factor alpha, interferon gamma and granulocyte macrophage colony stimulating factor levels in juvenile chronic arthritis. PMID- 10371289 TI - Prevalence of hypothyroidism among Arabs with rheumatoid arthritis. PMID- 10371290 TI - Etoposide in Wegener's granulomatosis. PMID- 10371291 TI - Allergic pancytopenia to trimethoprim-sulphamethoxazole for Pneumocystis carinii pneumonia following methotrexate treatment for rheumatoid arthritis. PMID- 10371292 TI - An unusual case of pigmented villonodular synovitis of the spine: benign aggressive and/or malignant? PMID- 10371293 TI - High serum level of macrophage-colony stimulating factor (M-CSF) in adult-onset Still's disease. PMID- 10371294 TI - Microscopic polyangiitis associated with antiphospholipid syndrome. PMID- 10371295 TI - Giant cell arteritis in a patient with limited cutaneous systemic sclerosis. PMID- 10371296 TI - Silastic prostheses--a forgotten cause of lymphadenopathy in rheumatoid arthritis. PMID- 10371297 TI - Increased antibody responses to Klebsiella serotypes K26, K36, and K50 in patients with ankylosing spondylitis. PMID- 10371298 TI - Acroparaesthesia--a typical finding in vitamin D deficiency. PMID- 10371299 TI - Establishment and characterization of normal and initiated hamster tracheal epithelial cell system. AB - The development and characterization of normal hamster tracheal epithelial (HTE) cell system and its initiated subline is described in the present study. Normal HTE cells grew in a monolayer, had a stable diploid karyotype, were anchorage dependent and non-tumorigenic. The presence of desmosomal attachments and keratin filaments confirmed the epithelial nature of these cells. An initiated subline DTC8 was isolated after treatment of HTE cells with a suboptimal dose of 9,10 dimethyl-1,2-benz[a]anthracene (DMBA). These DTC8 cells grew in a monolayer, had a higher growth rate and saturation density, were weakly anchorage independent and non-tumorigenic. Treatment of DTC8 cells with 100 ng 12-O-tetradecanoyl phorbol-13-acetate (TPA), resulted in transformation of these cells which then showed anchorage independent growth on semisolid agar and formed tumours in 85% animals. As DTC8 cells showed heterogeneity in chromosome number, they were further cloned by the limiting dilution method using gamma-irradiated hamster embryonic fibroblasts as a feeder layer. The clone H7I, isolated among these clones had all the properties of initiated cells. PMID- 10371300 TI - The need for dynamic methods for measuring cell cycle perturbations: a study in radiation-treated lymphoblastoid cell lines of varying p53 status. AB - Reports on the p53-related cell cycle and apoptotic responses of EBV-transformed lymphoblastoid cell lines to DNA damage have led to some confusion. This may be due to differences in the nature of the specific p53 mutations under examination, but it can also be partly attributed to methodological and analytical problems (e.g. the inappropriate use of static DNA histograms for cell cycle analysis). Taking seven lymphoblastoid cell lines derived from both normal individuals and Li-Fraumeni Syndrome/Li-Fraumeni-Like (LFS/LFL) patients of differing p53 status, we completed a detailed study of radiation-induced cell cycle perturbations. Using BrdUrd pulse labelling and flow cytometry it was found that, regardless of p53 status, the cells did not arrest in G1 despite all of the lines showing p53 upregulation 3 hours postirradiation. The irradiated cells did, however, show a general slowing both in S-phase entry from G1 and in movement through S-phase. These facts would not have been apparent from the analysis of static DNA histograms. The problems with the use of static methods to assess changes in the dynamics of cell cycle progression apply not only to studies involving EBV transformed cell lines, but also to a wide range of investigations into the molecular control of cell proliferation. PMID- 10371301 TI - Comparison of methods based on annexin-V binding, DNA content or TUNEL for evaluating cell death in HL-60 and adherent MCF-7 cells. AB - HL-60 and MCF-7 cells were treated with 0.15 microM camptothecin (CPT) or with the solvent dimethylsulfoxide (DMSO) for the controls, for 2, 3 and 4 h or for 24, 48 and 72 h, respectively. The apoptotic index (AI) was then evaluated in parallel by the following flow cytometric methods: (1) double staining of unfixed cells with fluoresceinated annexin V and propidium iodide (PI), this after detachment by trypsinization in the case of MCF-7 cultures; (2) prefixation in 70% ethanol, extraction of degraded, low molecular weight DNA with 0.2 M phosphatecitrate buffer and analysis of the DNA content stained with PI; (3) TUNEL, i.e. labelling of DNA strand breaks with biotin-dUTP, followed by staining with streptavidin-fluorescein and counterstaining with PI. In HL-60 cells, the three methods gave similar results for the AI (3-4% in the controls and at 2 h of CPT treatment, and 35-43% at 3 and 4 h after CPT). This indicates that CPT induced membrane alteration and DNA fragmentation occurred concomitantly in those cells. For MCF-7 cells, CPT-induced apoptosis developed more slowly, the AI, whether based on annexin V or on DNA content, remained unchanged at 24 h, then was increasing to 8% at 48 h and to 25% at 72 h of treatment. In these cells, the TUNEL index did not increase prior to 72 h, and the increase was minor (up to 9% vs. 2-3% in the controls) at 72 h of the treatment. This indicates that in MCF-7 cells DNA strand breaks cannot be effectively labelled, which may be due to inaccessibility of 3'-OH ends in the breaks to exogenous terminal deoxynucleotidyl transferase. The mechanism of endonucleolytic DNA fragmentation thus may be different, depending on the cell type. PMID- 10371302 TI - Oscillating growth patterns of multicellular tumour spheroids. AB - The growth kinetics of 9L (rat glioblastoma cell line) and U118 (human glioblastoma cell line) multicellular tumour spheroids (MTS) have been investigated by non-linear least square fitting of individual growth curves with the Gompertz growth equation and power spectrum analysis of residuals. Residuals were not randomly distributed around calculated growth trajectories. At least one main frequency was found for all analysed MTS growth curves, demonstrating the existence of time-dependent periodic fluctuations of MTS volume dimensions. Similar periodic oscillations of MTS volume dimensions were also observed for MTS generated using cloned 9L cells. However, we found significant differences in the growth kinetics of MTS obtained with cloned cells if compared to the growth kinetics of MTS obtained with polyclonal cells. Our findings demonstrate that the growth patterns of three-dimensional tumour cell cultures are more complex than has been previously predicted using traditional continuous growth models. PMID- 10371303 TI - Correlation between cholesterol esterification, MDR1 gene expression and rate of cell proliferation in CEM and MOLT4 cell lines. AB - A positive correlation between cholesterol esterification and growth rate potential was previously found in our laboratory during the growth of CEM and MOLT4 lymphoblastic cells. In the current study, we investigated whether the rates of cholesterol esters synthesis correlate with changes of acyl CoAcholesterol acyltransferase (ACAT) mRNA levels and of other genes implied in cholesterol biosynthesis and uptake, such as 3-hydroxy-3-methylglutaryl-CoA (HMGCoA) reductase and low density lipoprotein (LDL) receptor. The results showed that the more rapid growing CEM cells had lower levels of expression of HMGCoA reductase and LDL receptors compared to MOLT4. By contrast, ACAT mRNA levels were higher in CEM cells, further supporting the concept of a possible involvement of cholesterol esters in the regulation of cell growth and division. In this study, high levels of cholesterol esterification and of expression of ACAT gene were also associated with a markedly increased expression of multidrug resistance (MDR1) gene, suggesting that MDR1 activity might contribute to regulate the rate of cell growth and division by modulating intracellular cholesterol ester levels. PMID- 10371304 TI - Induction of apoptosis by hinokitiol, a potent iron chelator, in teratocarcinoma F9 cells is mediated through the activation of caspase-3. AB - Hinokitiol, a potent iron chelator, has been reported to induce differentiation in teratocarcinoma F9 cells with a reduction of viable cells. In this study, we examined the steps leading to eventual cell death by hinokitiol during differentiation. Hinokitiol induced DNA fragmentation of F9 cells in a concentration- and time-dependent manner. This effect was also observed in a cell free system using the nuclei from intact cells and the cytosols from hinokitiol treated cells. In contrast, hinokitiol methyl ether and hinokitiol-Fe (III) complex, which are deficient in iron-chelating activity, showed no DNA fragmentation activity in both cell culture and cell-free systems. These results suggest that iron deprivation by hinokitiol may be involved in the induction of apoptosis of F9 cells. Caspase-3, one of the key enzymes in the apoptotic cascade, was specifically activated by hinokitiol treatment, but not by the other two derivatives. In addition, its specific inhibitor, benzyloxycarbonyl-Val-Ala Asp-fluoromethyl ketone, strongly blocked hinokitiol-induced DNA fragmentation. These results indicate that iron deprivation by hinokitiol can induce apoptosis of F9 cells through the activation of caspase-3. PMID- 10371305 TI - Do you CARE? A national register for cleft lip and palate patients. Craniofacial Anomalies Register. PMID- 10371306 TI - Mandibular condyle fractures: a consensus. AB - A consensus was obtained following a two-day international conference to review the management of mandibular condyle fractures. Whilst areas of disagreement still exist, there are many areas of agreement. It is hoped this editorial will stimulate debate leading to internationally accepted guidelines. PMID- 10371307 TI - Occlusal outcome in patients undergoing orthognathic surgery with internal fixation. AB - Fifty consecutive patients undergoing orthognathic surgery with internal fixation (IF) were studied retrospectively with a weighted Peer Assessment Rating (PAR) Index to assess occlusal outcome at the end of all active treatment, and compared with 50 patients who had undergone treatment for malocclusion by orthodontic means alone. In the surgically treated patients, the mean percentage reduction in the weighted PAR Index was 83% and 31 out of 38 patients (82%) were 'greatly improved'. This implies a high standard of treatment in terms of the occlusal outcome. There was no difference in the proportion of patients having a final weighted PAR Index of less than 10 and no significant difference in the final weighted PAR Index between the two groups. This suggests that the occlusal outcome is no different whether patients undergo orthognathic surgery or orthodontic treatment alone, and that excellent occlusal results can be achieved in patients undergoing orthognathic surgery with internal fixation. PMID- 10371308 TI - Digital imaging in the assessment of facial deformity. AB - A combination of digital imaging and conventional radiographic techniques has been used to obtain a life-size composite image of the facial soft tissue profile, the skull, and the teeth, anatomically superimposed on each other. The system overcomes major problems associated with assessment of facial deformity such as magnification, the exposure of photographs and radiographs from different positions, radiographs with overexposed soft tissue profiles, lengthy procedures in photographic studios, and, in addition, enables a degree of partial automation in the management of patients. PMID- 10371309 TI - An adjustable bone fixation system for sagittal split ramus osteotomy: preliminary report. AB - After sagittal split osteotomy, osteosynthesis for stabilization of the fragments is usually performed by means of rigid or semirigid fixation to avoid maxillomandibular fixation (MMF). Sometimes, this leads to malpositioning of the fragments with resulting 'immediate relapse' or disturbances of the temporomandibular joints (TMJs) such as progressive idiopathic condylar resorption, which is the worst effect. We have therefore developed a new bone fixation system that allows three-dimensional adjustment even after fixation of the fragments. The special configuration of the system avoids intraoperative dislocation of the mandibular condyle. We have used the method successfully in 40 patients, 27 of whom had mandibular retrognathism and 13 of whom had mandibular prognathism. In particular, there has been no evidence of resorptive changes. There were no postoperative complications. PMID- 10371310 TI - Ferromagnetic MRI artifact secondary to a previous mandibular modified condylotomy. PMID- 10371311 TI - Jaw fractures in Enugu, Nigeria, 1985-95. AB - A retrospective analysis of 900 patients with jaw fractures of the facial bones during the period January 1985 - December 1995 indicated that 747(83%) resulted from road traffic accidents, 75(8.4%) from interpersonal violence, 39(4.3%) from accidents during sporting events, and 36(4%) from occupational accidents, while the causes of 3(0.3%) were not stated. The left side of the face was affected more often than the right. The mandible was twice as likely to be fractured as the zygomaticomaxillary complex. The symphysis-body-angle and the condylar region were the most common sites of fracture of the mandible, while the zygoma was the area most often affected in the middle third of the face. Most maxillofacial fractures occurred in the age group 21-30 years, and the lowest among those over 60. Three times as many men were affected as women. We conclude that there is high incidence of fractures of the facial bones caused by traffic accidents in our environment and, in all age groups, men were more likely to be affected than women. PMID- 10371312 TI - Policy of consultant oral and maxillofacial surgeons towards removal of miniplate components after jaw fracture fixation: pilot study. AB - A pilot study was undertaken to find out the policy of the 23 consultant oral and maxillofacial surgeons in the West Midlands towards removal of miniplates after jaw fractures had healed. All 23 replied. Two consultants did not use miniplates; the other 21 respondents used titanium systems, and two also used a stainless steel system. None of the 21 respondents routinely removed all miniplates. The estimated total of miniplates removed after the fracture had healed ranged between 5% and 40% (mode 5%). The main indications for removal were wound infection or dehiscence, before construction of a prosthesis, patients' concern about permanent retention of an implant, and thermal conductivity. We conclude that miniplates and screws are removed mainly to treat symptoms caused by the implants. PMID- 10371313 TI - Fractured surgical instruments located using radiographs of the suction bottle. AB - This paper describes the use of radiographs of suction bottle receptacles for locating elusive fragments of surgical instruments that have fractured per operatively. PMID- 10371314 TI - The intermaxillary screw: a dedicated bicortical bone screw for temporary intermaxillary fixation. PMID- 10371315 TI - The pearl steel wire: a simplified appliance for maxillomandibular fixation. PMID- 10371316 TI - Closure of radial forearm free flap donor site with local full-thickness skin graft. AB - A triangular shaped full-thickness skin graft harvested adjacent to the donor site of the radial forearm flap, as originally described by Liang et al, has successfully been used in seven consecutive patients for coverage of the donor site of the radial forearm free flap. In all patients this resulted in a robust coverage with no late wound breakdown and an aesthetic appearance far superior to split-thickness skin-graft coverage. We recommend this technique which is feasible in the majority of cases and reduces both donor site and graft site morbidity of the radial forearm flap. PMID- 10371318 TI - Arteriovenous fistula after temporomandibular arthroscopy. AB - Temporomandibular joint arthroscopy has been associated with various vascular injuries including haemorrhage, pseudoaneurysm and fistula. We describe the endovascular balloon embolization of a traumatic superficial temporal arteriovenous fistula that complicated TMJ arthroscopy. We conclude that suspected vascular injuries after this procedure should be investigated by arteriography, and that embolization is a safe and effective treatment for superficial temporal artery fistulas. PMID- 10371317 TI - Temporomandibular joint ankylosis: review of thirty-two cases. AB - I have reviewed aetiology, sex, age at time of treatment, clinical features, radiographic findings, anaesthetic techniques, surgical treatment, complications, and results in 32 patients with ankylosis of the temporomandibular joint. Trauma and infection were the commonest causes of ankylosis: 50% and 41%, (n = 13), respectively. The 21-30 year age group had the most trauma cases. Twenty (63%) of the patients presented with bilateral ankylosis. Failing to do jaw-opening exercises was the main cause of relapse. PMID- 10371319 TI - Technique for harvesting tibial cancellous bone modified for use in children. AB - We present modifications of the technique of harvesting adult tibial cancellous bone that enables its safe and reliable use in children. We studied 15 children undergoing secondary grafting of alveolar clefts radiographically and clinically and compared them with a similar group of children who had bone harvested from the iliac crest. Duration of hospital stay (mean 1, range 1-2 compared with 3, range 2-4) analgesic requirements (duration 1-4 days compared with 1-3 weeks) were all considerably less when bone was taken from the proximal tibia and mobility was considerably improved (able to walk on day 1 compared with restricted walking for 1-3 weeks. PMID- 10371320 TI - Sensory disorders after separation of the nasopalatine nerve during removal of palatal displaced canines: prospective investigation. AB - During a prospective study after separation of the nasopalatine nerve at the foramen incisivum during exposure or removal of impacted and palatal displaced maxillary canines, 59 patients were examined neurologically for 4 weeks postoperatively over an investigation period of 18 months. During the first week after the operation, subjective as well as objective sensory disorders were found in all of the patients, but after 4 weeks at the most no neurological deficit could be detected in any patient. PMID- 10371321 TI - Tetracycline sclerotherapy for the treatment of recurrent pooling of plasma in the submandibular tissue space: case report. AB - We present an unusual case of recurrent swelling after removal of the submandibular and sublingual salivary glands which was found to be the result of a collection of plasma. This was successfully treated by an injection of tetracycline to induce sclerosis. PMID- 10371322 TI - Polyvinylsiloxane dental bite registration material used to splint a composite graft of the nasal rim. AB - We describe the construction of a nostril splint made from heavy body silicone based dental impression material. This bio-compatible material supported a large chondrocutaneous auricular graft during early healing. The splint immobilized and maintained the shape of the alar rim and was further used nightly during the expected period of wound contraction. PMID- 10371323 TI - Electrocautery-ignited endotracheal tube fire: case report. AB - The risk of fire in the airway associated with laser surgery is well known, but there are reports of endotracheal tube fires ignited by electrocautery, particularly during pharyngeal surgery or tracheostomy or both. This uncommon complication has potentially devastating consequences. Surgeons undertaking these procedures should be aware of this complication and be familiar with measures to avoid them. We present a case report of an electrocautery-ignited endotracheal tube fire during an elective tracheostomy, which resulted in the patient's death. PMID- 10371325 TI - Re: Devani et al. Dental extractions in patients on warfarin: is alteration of anti-coagulant regime necessary? PMID- 10371324 TI - Postanginal septicaemia with external jugular venous thrombosis: case report. AB - Postanginal septicaemia is a syndrome of anaerobic septicaemia, septic thrombophlebitis of the internal jugular vein, and metastatic infections, that follows a localized infection in the area drained by the large cervical veins. The syndrome was well-known and often fatal in the preantibiotic era. It is now rather rare, presumably as a result of the almost routine use of prophylactic antibiotics. The symptoms are classic, and it should be suspected in any case where septicaemia and metastatic lesions are preceded by a head and neck infection. We report a case that is typical, except that branches of the external jugular vein were thrombosed. To our knowledge this has not been reported previously. PMID- 10371326 TI - Osteoradionecrosis and hyperbaric oxygen. PMID- 10371328 TI - Delayed tracheal extubation in oral and maxillofacial surgery. PMID- 10371327 TI - Madelung's disease, an asymmetric presentation. PMID- 10371329 TI - Non-ossifying fibroma of the mandible: report of a case. PMID- 10371331 TI - Histological subtypes of pleomorphic adenoma and age-frequency distribution. PMID- 10371330 TI - Osteomyelitis of the mandible in HIV infection. PMID- 10371332 TI - Standard of proof for medical negligence. PMID- 10371333 TI - Orthodontic bracketing: an alternative for intermaxillary fixation in mandibular fracture. PMID- 10371334 TI - A simple index using video image analysis to predict disease outcome in primary breast cancer. AB - Image analysis was used to investigate the prognostic significance of immunostaining for oestrogen receptor (ER), p53 tumour-suppressor protein and tumour cell proliferation (MIB-1) in a random cohort of 200 primary breast cancer patients with between 4 and 7 years of clinical follow-up. Image measurements of the percentage of immunopositive cancer cell nuclei (% positive nuclear area) were recorded for the above tumour features for each patient. Assessment of relative risk using Cox's univariate analysis indicated that tumour size, number of cancer-involved nodes, MIB-1 and ER % positive nuclear area were significantly associated with breast cancer disease outcome, i.e., relapse-free survival and overall survival. In multivariate analysis, tumour size, number of involved nodes, ER and MIB-1 % positive nuclear area were retained as independent predictors of prognosis, depending on the image measurement cut-point used. A prognostic model, which can be used without reference to nodal involvement, was constructed for tumour size, ER cut-point of 30% positive nuclear area and MIB-1 cut-point of 10% positive nuclear area. Kaplan-Meier analysis of this image-based prognostic index identified 4 risk groups with predicted 5-year overall survival rates of 93%, 83%, 76.7% and 61.5%. We conclude that image measurements of ER and proliferative rate can be combined with tumour size to construct a prognostic index which reliably predicts disease outcome in primary breast cancer without knowledge of the nodal status of the patient. PMID- 10371335 TI - Prognostic value of CD44 variant expression in primary breast cancer. AB - CD44 is a family of cell surface transmembrane glycoproteins members which differ in the extracellular part by sequences derived by alternative splicing of 10 variant exons (v1-v10). CD44 proteins containing such variant sequences have been implicated in tumor metastasis formation. Here, we have evaluated the expression of CD44 variants by immuno-histochemistry in primary breast cancer samples of 237 node-negative and 230 node-positive patients. For the analysis of samples derived from node-negative patients, the exon-specific antibodies used were DIII, vff7 and vff18 (v6), vff17 (v7/v8), fw11.24 (v9) and vff16 (v10). With the different antibodies which recognize v6 epitopes, the majority of tumors were positively stained (> or = 65% of the tumors) with varying intensities. Thirty-nine percent of the tumors were positively stained with the antibody vff16, and approximately half of the tumors with the antibodies vff17 and fw11.24. The expression of CD44 v6 epitopes in tumors from node-negative patients was associated with a favorable prognosis, both upon univariate and multivariate analysis. The expression of CD44 v7/8, v9 or v10 epitopes was not significantly related with relapse-free survival. Samples from node-positive patients were only examined with the antibodies vff7, vff17 and vff18. The staining with none of these antibodies was correlated with the length of relapse-free survival of the patients. Our data suggest that, generally, the usefulness of knowledge of CD44 variant expression is of limited value for assessing the risk of relapse in patients with primary breast cancer. However, the expression of exon v6 of CD44 may be a marker to identify patients with a relatively favorable prognosis in node-negative patients. PMID- 10371336 TI - No evidence for germline PTEN mutations in families with breast and brain tumours. AB - Germline mutations of the PTEN gene are involved in Cowden disease, a genetic condition associated with an increased risk of breast cancer. Further somatic PTEN mutations have been found in glioblastomas and to a lesser extent in meningiomas. Therefore, PTEN germline mutations were searched for in a series of 20 unrelated women with breast cancer who also had a personal or familial breast brain tumour history. Inclusion criteria were 1. family history of breast cancer; 2. absence of germline BRCA1 and p53 mutation; and 3. at least one case of brain tumour (glioblastoma, meningioma, or medulloblastoma) in either the index case or one of their first or second degree relatives. Any stigmata of Cowden disease was an exclusion criteria. Screening of the PTEN gene for point mutations or small rearrangements were performed using the denaturing gradient gel electrophoresis method on the 9 coding exons. No disease-associated mutation of the PTEN gene has been detected in our series. It is, thus, unlikely that PTEN is a significant BRCA predisposing locus. However, one might ask whether breast cancer cases resulting from germline PTEN mutation could occur without any mammary histological feature of Cowden disease. PMID- 10371338 TI - Expression of cell cycle markers in colorectal carcinoma: superiority of cyclin A as an indicator of poor prognosis. AB - Our aim was to analyze the relationship between the proliferative activity of cancer cells, assessed using some cell cycle markers, and clinicopathological factors in colorectal carcinoma patients. Immunostaining for Ki-67 (pan-cell cycle marker), cyclin D1 (G1-phase marker) and cyclin A (S- to G2-phase marker), and in situ hybridization for histone H3 mRNA (S-phase marker) were carried out. Immunoreactivity was evaluated semiquantitatively using a scoring system to calculate a staining index (SI). The expression of cyclin D1, histone H3 mRNA and cyclin A correlated significantly with Ki-67 antigen expression. The SIs of Ki 67, cyclin A and histone H3 mRNA were significantly higher in patients > or = 65 years of age than in those < 65. The SIs of Ki-67 and cyclin D1 in poorly differentiated adenocarcinomas were significantly higher than in the other tumor types. Furthermore, the SI of cyclin D1 in carcinomas with lymph node metastasis was higher than in carcinomas without metastasis and was higher in advanced carcinomas than early carcinomas. The overall survival was significantly lower in patients with cyclin A overexpression than in those without. Multivariate analysis indicated that cyclin A overexpression is an independent prognostic factor in patients with colorectal adenocarcinoma. Our results indicate that cyclin D1 overexpression correlates with poor adenocarcinoma differentiation and tumor progression, and cyclin A overexpression is a superior indicator of poor prognosis compared with the other cell cycle markers tested. PMID- 10371337 TI - Glutathione-S-transferase polymorphisms and risk of squamous-cell carcinoma of the head and neck. AB - Differences in genetic susceptibility to tobacco-induced carcinogenesis appear to modulate an individual's risk of squamous-cell carcinoma of the head and neck (SCCHN). Risk for SCCHN may be associated with the null alleles of the carcinogen metabolizing genes glutathione-S-transferase (GST) T1 and GSTM1. In this study, we evaluated the association between GSTM1 and GSTT1 null genotypes and risk of SCCHN in a matched case-control study of 162 patients with SCCHN and 315 healthy controls. Our results showed that 53.1% of cases and 42.9% of controls were null for GSTM1, whereas 32.7% of cases and 17.5% of controls were null for GSTT1 (p < 0.05 and p < 0.001, respectively). Furthermore, 19.8% of cases but only 7.9% of controls were null for both genes (p < 0.001). Multivariate analysis using logistic regression models, including age, sex, ethnicity, smoking status, alcohol status and GST genotypes, showed that both of these genotypes remained independent risk factors for disease [adjusted odds ratios (ORs) = 1.50 and 2.27, respectively; 95% confidence intervals (CIs) = 1.01-2.23 and 1.43-3.60, respectively). When the genotypes were divided into neither null, either null or both null, there was a dose-response relationship (adjusted OR = 1.50, 95% CI = 0.98-2.30) for the either-null group and (adjusted OR = 3.64, 95% CI = 1.94-6.84) for the both-null group (p < 0.001, trend test). Our findings suggest that the GSTM1 and GSTT1 null genotypes are independent risk factors for SCCHN and markers for genetic susceptibility to tobacco-induced carcinogenesis. PMID- 10371339 TI - Heat-shock-protein-27 (hsp27) expression in ovarian carcinoma: relation in response to chemotherapy and prognosis. AB - Heat-shock protein 27 (hsp27) is one of the small heat-shock proteins. Its expression in ovarian- and breast-cancer cell lines has been associated with resistance to cisplatin and doxorubicin. In addition, hsp27 expression appears to facilitate cellular growth, differentiation and motility. In several human carcinomas, hsp27 expression might also be related to worse prognosis. The aim of this study was to evaluate the prognostic value of hsp27 expression in patients with ovarian carcinoma in relation to their response to chemotherapy and overall survival. Hsp27 expression was assessed by immunohistochemistry in 77 patients with ovarian carcinoma stage IC-IV. All patients received cisplatin- and doxorubicin-based chemotherapy and had long-term follow-up. In 30 patients, paired tumour samples were available, obtained before and after chemotherapy. Hsp27 immunostaining was positive in 86% of patients before and in 72% of patients after chemotherapy. Hsp27 expression was not related to any clinicopathologic factor, including previously determined p53 expression. Univariate analysis showed that, in stage-III and -IV patients, younger age, no residual tumour after first laparotomy, < or = 1 litre ascites, response to first line chemotherapy and absence of hsp27 expression were associated with longer median progression-free survival. However, in multivariate analysis, only age, ascites and response to chemotherapy retained independent prognostic value. PMID- 10371340 TI - Expression of Fas (CD95/APO-1) and Fas ligand in lung cancer, its prognostic and predictive relevance. AB - In order to examine whether or not the expression of the apoptosis-related receptor Fas (CD-95/APO-1) and its ligand (FasL) has relevance for patient survival, immunohistochemistry was used to analyze the proteins of both factors in 164 non-small cell lung carcinomas. Patients with Fas-positive tumors exhibited significantly longer survival times than patients with Fas-negative carcinomas. In contrast, FasL did not significantly influence patient survival time. A multivariate analysis of clinical and biological factors indicated that lymph node status and Fas expression were significant prognostic factors. Carcinoma patients who were negative for both Fas and FasL had a significantly higher incidence of lymph node involvement than did carcinoma patients who were positive for Fas and FasL. Carcinomas that were positive for Fas and FasL demonstrated a greater sensitivity to doxorubicin in vitro. PMID- 10371341 TI - Expression of cyclins and cyclin-dependent kinases in smooth muscle tumors of the uterus. AB - To investigate the cell cycle regulatory mechanisms involved in the growth of smooth muscle tumors, we studied the expression of Ki-67, cyclins E and A, and their catalytic partners, the cyclin-dependent kinases cdk2 and cdc2 by using tissue specimens from benign and malignant smooth muscle tumors. These included 20 cases of usual leiomyoma (UL), 18 of cellular leiomyoma (CL), 8 of bizarre leiomyoma (BL), 8 of uncertain malignant potential tumors (UMP) and 20 of leiomyosarcoma (LMS). The proliferation rate detected by Ki-67 was low in normal myometrium and leiomyomas (UL, CL and BL), but it was markedly increased in LMS. The expression of the cyclins (E and A) and cdks (cdk2 and cdc2) was also low in normal myometrium and leiomyomas. However, the expression of these factors was markedly increased in LMS. In addition, a survival analysis using Log-rank test, revealed that LMSs with positive staining for cyclin A and with diffusely staining for cyclin E were associated with significantly shorter survival. Our results suggest that expression of cyclins and cdks may be involved in the growth control of uterine smooth muscle tumors. PMID- 10371342 TI - Expression of mucin 1 (MUC1) in esophageal squamous-cell carcinoma: its relationship with prognosis. AB - Using 2 anti-mucin 1 (MUC1) monoclonal antibodies (MAbs), DF3 and BCP8, we examined MUC1 expression immunohistochemically in 192 esophageal squamous-cell carcinomas (SCCs). In normal squamous epithelium of the esophagus, DF3 was not expressed, but BCP8 was expressed on the cell membrane, mainly in the surface layer. In esophageal SCCs, DF3 and BCP8 were expressed mainly on the cell membrane of SCC cells, but also in the cytoplasm in several cases. To analyze the correlation of MUC1 expression and the prognosis of the patients, the 192 cases were divided into 2 groups: high-expression group (HEG, > 50% of the neoplastic cells stained) and low-expression group (LEG, < 50% of neoplastic cells stained). DF3-HEG (24 patients) showed a significantly poorer survival rate than DF3-LEG (168 patients), whereas there was no significant difference in survival between BCP8-HEG (43 patients) and BCP8-LEG (149 patients). Also, in the analysis of 162 patients with advanced stage (submucosal or deeper invasion) to exclude the influence of low expression of DF3 and BCP8 in 30 patients with early stage (up to the level of muscularis mucosae), DF3-HEG (24 patients) showed significantly poorer survival than DF3-LEG (138 patients), whereas there was no significant difference in survival between BCP8-HEG (42 patients) and BCP8-LEG (120 patients). The results of our study on esophageal SCC suggest that the expression of sialyl oligosaccharides detected by DF3 is related to poor prognosis. PMID- 10371343 TI - BRCA1 expression levels predict distant metastasis of sporadic breast cancers. AB - The role of BRCA1 in progression of sporadic breast cancers has to date been equivocal, although preliminary studies on small numbers of samples have suggested an association between expression levels of this gene and acquisition of an invasive phenotype. We have further reasoned that loss of oestrogen receptor positivity may have a detrimental effect on BRCA1 expression. In order to test this hypothesis and extend earlier investigations we have applied a sensitive RT-PCR procedure to determine the associations between BRCA1 expression and a variety of clinical parameters in a sample cohort derived from sporadic breast tumour specimens. We have established that BRCA1 and ER mRNA expression are closely associated (p=0.013), indicating a possible functional relationship between these 2 genes. We have further identified an association between low levels of BRCA1 expression and acquisition of distant metastasis in sporadic disease (p=0.019). In light of our findings, we suggest that suppression of BRCA1 has a role to play in progression of a significant fraction of sporadic breast cancers and may additionally prove to be a useful, novel, prognostic marker for this disease type. PMID- 10371344 TI - Novel indications for BRCA1 screening using individual clinical and morphological features. AB - Since there is a lack of common family profile among BRCA1-gene carriers, and since the risk of being a mutation carrier is not limited to women with a family history of breast or ovarian cancer, multivariate statistical analysis using the logistic-regression model was carried out, to discriminate between sporadic cases and BRCA1-breast cancers (BRCA1-BCs), especially when information about the family history of breast/ovarian cancer and ethnicity are irrelevant or unavailable, in order to offer specific medical treatment to this population. We examined 32 BRCA1-BCs selected at cancer genetic clinics and 200 consecutive controls without family history of breast cancer for age at onset and current morphological parameters. Following the multivariate analysis, 3 parameters only, namely, early age at cancer onset [odds ratio (OR) for each year = 1.16; p < 0.0001], estrogen-receptor negativity (OR = 5.7; p = 0.01) and poor differentiation (OR = 5; p = 0.03) were found significant factors for predicting BRCA1-carrier status. The expected impact in BRCA1 screening of our model was estimated using data on 5700 breast-cancer cases from a hospital-based registry. Only 50 and 15% of tumours with early age at onset below 35 years present one or the other 2 discriminant parameters respectively. Consequently, whereas the probability of finding a BRCA1 mutation is rated low (6.2%) when the sole criterion of early onset up to the age of 35 years is used, based on our model, in the sub-group of women with a tumor that is both estrogen-receptor-negative and poorly differentiated the mutation-detection rate is predicted to be above the 10% chance level recommended by the ASCO guidelines. This sub-group of women, representing about 1% of all breast-cancer cases in Western countries, consequently deserves to be tested. PMID- 10371346 TI - c-erbB-2 over-expression in amplified and non-amplified breast carcinoma samples. AB - We investigated c-erbB-2 oncogene amplification and over-expression in 79 invasive breast carcinoma samples using fluorescence in situ hybridization (FISH) and immunohistochemistry, with the aim of studying relationships between neoplasms over-expressing c-erbB-2 with or without amplification and bio pathological parameters used in clinical breast cancer. Nineteen samples showed amplification, and all of these were positive by immuno-histochemistry. Moderate or intense immunostaining was present in a further 22 samples without c-erbB-2 amplification and was not related to any increased number of c-erbB-2 signals: 15 samples exhibited chromosome 17 polysomy, 3 monosomy and 4 no FISH abnormalities. Thirty-eight samples were immunonegative: 18 exhibited chromosome 17 polysomy, 9 monosomy and 11 no alterations. Samples having c-erbB-2 over-expression associated with amplification showed DNA aneuploidy and hormonal receptor loss to a greater extent than those expressing c-erbB-2 without amplification or immunonegative samples (chi2 test, p = 0.007, 0.008 and 0.008, respectively). The proliferation rate, detected by Ag-NOR staining, was highest in amplified samples (Kruskal Wallis test, p = 0.009). Our results indicate that tumours showing both c-erbB-2 over-expression and amplification exhibit more aggressive biological characteristics than those with only over-expression or immunonegative tumours. Since both c-erbB-2 amplification and over-expression have been related to negative responses to chemotherapy and poor prognosis, these differences might have clinical implications. The combination of FISH and immuno-histochemistry may be helpful to achieve this aim. PMID- 10371345 TI - Cyclin-D1 expression in human renal-cell carcinoma. AB - Cyclin-D1 over-expression represents one of several common alterations in the G1 S transition associated with malignancies. Conclusive evidences indicate that cyclin D1 is a proto-oncogene and the gene is amplified or rearranged in different tumour types. Since very little is known about aberrations in the G1-S transition in human renal-cell carcinoma (RCC), we have characterized the expression of cyclin D1 in 80 human renal-cell carcinomas and 12 normal kidney cortex tissues using Western blotting. The cyclin-D1-protein content varied considerably and 75% of the tumours expressed higher levels than normal kidney cortex, in contrast to 25% of the tumours either lacking cyclin D1 or with low protein levels. Although it is difficult to define aberrant expression of cyclin D1, the results might indicate that the proto-oncogene was activated in a sub-set of RCC. It is also possible that low expression of cyclin D1 represents an aberrant down-regulation of the protein. Immunohistochemical assessment of cyclin D1 in a sub-set of the tumours showed large variations in the fraction of cyclin D1-positive cells, supporting the Western-blot analyses. Surprisingly, cyclin-D1 expression did not correlate with proliferation determined by Ki-67-antigen expression or S-phase analyses. In non-papillary renal-cell carcinomas, high cyclin-D1 expression was associated with a diploid DNA profile and smaller tumour size, but there was no association between cyclin-D1 expression and tumour stage or nuclear grade. In nonpapillary tumours, high cyclin-D1 expression was further significantly associated with a better prognosis according to univariate and multivariate analyses (p = 0.005 and 0.002 respectively), as compared with highly aggressive tumours with low cyclin-D1 levels. PMID- 10371347 TI - Expression of c-erbB2 and p53 protein is similar in breast cancer from British and Japanese women. AB - In an attempt to explain the difference in outcome between British and Japanese women with breast cancer we have compared histopathological features, expression of c-erbB2 and p53 proteins and clinical outcome of 191 British (Anglo) women with 171 Japanese women treated between 1979 and 1980. The Japanese patients were significantly younger than the Anglo patients, while in premenopausal women the latter had significantly smaller tumors. The proportion of tumors expressing c erbB2 and p53 proteins was similar in both populations. c-erbB2 positivity was significantly associated with positive lymph node status and with poorly differentiated carcinomas. Duration of relapse-free and overall survival was significantly longer in the Japanese women than in the Anglo women. Women with c erbB2-negative tumors had a longer overall survival than women with c-erbB2 positive tumors and this difference was accentuated when patients were stratified according to country of origin. Japanese women with c-erbB2-negative tumors had the best outcome, whereas the Anglo women with c-erbB2-positive tumors had the worst. There was no relationship between p53 status and any histopathological features or clinical outcome. Differences in the expression of c-erbB2 and p53 do not explain the better outcome experienced by Japanese breast cancer patients. PMID- 10371349 TI - Molecular mediators of tumor angiogenesis: enhanced expression and activation of vascular endothelial growth factor receptor KDR in primary breast cancer. AB - The progression of breast cancer growth and its ability to metastasize are associated with the process of angiogenesis. In this study, we examined the protein expression of vascular endothelial growth factor (VEGF) and its specific and functional receptor KDR in human breast tissue. We investigated a total of 13 mammary carcinomas, 3 fibroadenomas, 5 specimens with fibrocystic breast disease as well as normal (adjacent to malignant) breast tissue using immunohistochemistry and Western blot analysis. In all carcinomas examined, functional KDR protein was present independent of tumor type, tumor stage and histological grade as demonstrated by tyrosine phosphorylation analysis of KDR. When malignant tissues were compared with their neighboring non-neoplastic regions, activated KDR was found to be expressed to a much higher extent within the malignant tissue samples. In fibroadenomas, KDR was barely detectable, whereas in fibrocystic breast disease KDR expression was variable. Immunostaining of KDR was localized to endothelium and epithelium of mammary ducts in malignant and benign breast tissue, while VEGF immunoreactivity was primarily found in the endothelium and also in tumor cells and macrophages. Our data demonstrate that KDR activation is enhanced in breast cancer in vivo and emphasize the functional role of VEGF and KDR in the development of malignant breast disease. PMID- 10371348 TI - Patient-specific mutation databases for oral cancer. AB - Development of databases, summarising the genetic events associated with oral squamous cell carcinoma (SCC), should increase our understanding of the molecular basis of these lesions. Additionally, databases will help establish whether different cancer subtypes show different growth characteristics, because the multistage carcinogenic process is different in the various tumour subtypes. This new knowledge may also provide new prognostic information, as these aberrations represent fundamental biological characteristics of each tumour. To assess the value of incorporating the results from loss of heterozygosity (LOH) analysis into patient-specific mutation databases, we have carried out microsatellite analysis with 52 polymorphic markers at 13 key chromosomal regions implicated in the pathogenesis of head and neck cancers. Altered expression of the Rb, p53 and DCC tumour suppressor genes has also been studied by immunohistology. Our results shed light on the different pathways that lead to cancer and reveal that a variety of different patterns of allelic imbalance (AI) were detected at all TNM stages, reflecting the different clinical behaviour that tumours classified as being of the same TNM stage may exhibit. Summarising the level of genetic damage as a fractional allelic loss (FAL) score and the presence of AI at 3p22-26, 3p14.3-12.1 and 9p21 was found to be a better predictor of outcome than the TNM system. This finding suggests that molecular data can be incorporated into conventional staging systems to provide more accurate prognostic information for this group of patients. PMID- 10371350 TI - Serum concentrations of squamous cell carcinoma antigen in patients with vulvar intraepithelial neoplasia and vulvar cancer. AB - Our aim was to evaluate whether serum concentrations of squamous cell carcinoma antigen (SCC-Ag) are an independent prognostic factor in patients with vulvar cancer. We measured SCC-Ag in pretreatment serum samples of 55 patients with squamous cell vulvar cancer, 30 patients with vulvar intraepithelial neoplasia (VIN) grade III and 50 healthy female controls. The results were compared with clinical data. Median serum concentrations of SCC-Ag in healthy female controls, patients with VIN III, and patients with invasive vulvar cancer were 0.5 (range 0.1 to 3.8) ng/mL, 0.5 (range 0 to 4.1) ng/mL and 1.6 (range 0.3 to 65) ng/mL, respectively (Mann-Whitney U test, p < 0.001). The 75% quantile of serum concentrations of SCC-Ag in patients with vulvar cancer was defined as cut-off level. Elevated pretreatment serum concentrations of SCC-Ag were significantly correlated with a shorter disease-free and overall survival (log-rank test, p=0.002; and p<0.001, respectively). A multivariate Cox regression model showed that serum concentrations of SCC-Ag are a prognostic factor of disease-free and overall survival independent of tumour stage (multivariate Cox regression model, p=0.03; and p=0.048, respectively). Pre-treatment serum concentrations of SCC-Ag were not correlated with tumour stage, histological grade and patients' age. In summary, our data indicate that serum concentrations of SCC-Ag may be an additional independent prognostic factor of disease-free and overall survival in patients with vulvar cancer. PMID- 10371351 TI - Detection of messenger RNA for the beta-subunit of chorionic gonadotropin in urinary cells from patients with transitional cell carcinoma of the bladder by reverse transcription-polymerase chain reaction. AB - We studied whether detection of messenger-RNA (mRNA) for the beta-subunit of chorionic gonadotropin (CGbeta) in urinary cells from bladder cancer patients could be used as a marker of disease activity. Sixty-eight urine samples from patients under follow-up for bladder cancer and 23 samples from patients with other malignancies and non-malignant surgical conditions, as well as 14 samples from healthy controls were analyzed. RNA was isolated from urinary cells collected by centrifugation. Reverse transcription-polymerase chain reaction (RT PCR) was used to detect CGbeta mRNA. The results were compared to those obtained by cystoscopy and urinary cytology. For comparison, we determined CG and CGbeta in serum and urine and the core fragment of CGbeta (CGbeta cf) in urine by immunofluorometric assays. CGbeta mRNA was detected in 29 of 68 urine samples from patients with a history of bladder cancer, whereas all 14 samples from healthy controls tested negative. Elevated levels of CGbeta were observed in serum in 18 of 45 bladder cancer patients, but the association with CGbeta mRNA was weak. However, CGbeta mRNA expression in the absence of detectable cancer also occurred in some conditions associated with cellular atypia such as urinary tract infection, instrumentation and certain therapies. There was a highly significant association between histologically verified transitional cell carcinoma of the bladder and CGbeta mRNA in urine (p = 0.0014), implying CGbeta mRNA expression in tumor tissue. We conclude that CGbeta mRNA is a potential new marker for monitoring of bladder cancer. Further studies are needed to evaluate whether it provides independent clinical information. PMID- 10371352 TI - Tumor-infiltrating dendritic cells in adenocarcinomas of the breast: a study of 143 neoplasms with a correlation to usual prognostic factors and to clinical outcome. AB - Dendritic cells (DC) are the most potent antigen-presenting cells, and induce antigen-specific immune responses. Infiltration of tumors by DC is thought to reflect the interaction between the host immune system and tumor cells. Tumor infiltrating DC (TIDC) are believed to evolve into tumor-antigen pulsed cells and then to migrate to local lymph nodes, where they activate anti-tumor immune responses. Indirect clinical evidence supporting this theory is provided by studies showing that high TIDC densities are associated with favorable prognosis in some tumor types. In the present study, we evaluated 143 primary breast adenocarcinomas for the presence of DC, using immunohistochemistry with the anti S100 protein antibody. We analyzed the relationship between the degree of infiltration by S100+ TIDC and the usual prognostic factors and clinical outcome. The results show that 42% of breast adenocarcinomas contain S100 TIDC. The number of S100+ TIDC varies according to the grade of tumors as follows: GRIII > GRII > GRI. A relationship is also found between S100+ TIDC and tumor size, lymph-node involvement, estrogen/progesterone receptor status and age. However, the presence of S100+ TIDC, even at the highest density, was not correlated with metastasis free survival or overall survival. PMID- 10371354 TI - Mutational analysis of the p73 gene in human breast cancers. AB - In primary breast cancer, mutations of the p53 tumor suppressor gene lead to loss of growth-suppressive properties and poor outcome. Recently, a p53-related gene, termed p73, has been cloned and its gene product possesses a function similar to p53. p73 has been mapped at chromosome 1p36.3, a region frequently deleted in breast cancer, neuroblastoma and other malignancies. To elucidate the functional significance of p73 in the oncogenesis of breast cancer, we have studied genetic alterations of p73 in tissue specimens obtained from 87 patients with primary breast cancer. Thirteen percent of informative cases showed loss of heterozygosity (LOH) at the p73 gene. However, there was no correlation between the p73 LOH and clinical features such as histopathological types, metastatic behavior or expression of estrogen or progesterone receptor. The levels of p73 transcript in primary breast cancer were not significantly different from those in normal breast tissue. Moreover, PCR-SSCP analysis failed to detect any missense or frameshift mutations in the p73 gene. Our observations suggest that allelic loss, expression levels and mutations of the p73 gene may not contribute to oncogenesis of primary breast cancers. PMID- 10371353 TI - Expression of p27 is associated with Bax expression and spontaneous apoptosis in oral and oropharyngeal carcinoma. AB - p27Kip1, a cyclin-dependent kinase inhibitor, is a negative regulator of the cell cycle, and apoptosis is a genetically encoded program of cell death. To clarify the relationship between the cell cycle and apoptosis, we investigated expression of p27, cyclin D1 and apoptosis-related proteins (p53, Bax, Bcl-2 and c-Myc) in 60 cases of oral and oropharyngeal squamous-cell carcinoma (SCC) using an immuno histochemical approach, and evaluated spontaneous apoptosis in vivo. Our most notable finding was that spontaneous apoptosis in the p27-positive group was significantly higher than that in the p27-negative group (p = 0.028). In addition, the percentage of p27-positive cells was clearly correlated with that of Bax-positive cells (gamma = 0.288, p = 0.028) and with that of cyclin D1 positive cells (gamma = 0.416, p = 0.002). Expression of p27 was inversely associated with the clinical stage of total tumor progression (p = 0.027). However, no correlation was found between p27 expression and the following parameters: gender, tumor size, lymph node metastasis, overall survival and disease-free survival. Our results give evidence that the action of the cell cycle regulator p27 is closely linked with apoptosis in clinical samples from patients and indicate that over-expression of p27 might induce apoptosis in cancer cells through elevation of Bax expression, thereby acting on tumor progression. PMID- 10371356 TI - Allelic loss on chromosome 6Q in primary prostate cancer. AB - Molecular genetic analyses of human prostate cancer (CaP) has revealed frequent loss of specific chromosome regions suggesting the presence of putative tumor suppressor gene(s) (TSG) on these chromosome loci whose inactivation may play a role in prostate tumorigenesis. To understand the role of 6q alterations in CaP, we have undertaken a comprehensive analysis of proximal 6q. Genomic DNA from tumor and normal prostate tissues from radical prostatectomy specimens of 38 patients were analyzed by polymerase chain reaction (PCR) for 13 polymorphic microsatellite loci on 6q. Allelic losses of 1 or more polymorphic loci were detected in 11 of 38 patients (29%). Six of 11 tumors showing any 6q deletion were found to have allelic losses at D6S1056 and D6S300 loci. Our results revealed a 1.5 megabase interval between D6S1056 and D6S300 at 6q16.3-21 as the minimal region of deletion, which may contain the putative TSG involved in prostate tumorigenesis. One of the tumor samples demonstrated homozygous deletion at a distal location D6S314 (6q23-24), suggesting another locus potentially associated with CaP. Although the relationship of 6q loss of heterozygosity (LOH) with various clinico-pathologic variables, i.e., cancer recurrence or pathologic stage, did not reveal a statistically significant association, the risk for 6q LOH to non-organ confined (pT3) disease was 5-fold higher than for organ confined disease. PMID- 10371355 TI - Modulation of interleukin-18 expression in human colon carcinoma: consequences for tumor immune surveillance. AB - The production in colon cancer of interferon-gamma (IFN-gamma), a type-1 T-helper (TH1) cytokine, is considered as a marker of good prognosis. We asked whether interleukin-18 (IL-18), which strongly induces IFN-gamma and regulates Fas ligand (Fas-L)-dependent cytotoxicity, may play a role in colon homeostasis, and if its expression was modulated in colon adenocarcinomas. We analyzed 14 specimens of colon adenocarcinomas, 6 of normal colon mucosa of the series, and 6 colon-tumor cell lines. The expression of IL-18, of ICE protease, involved in the processing of this cytokine, and of the downstream effectors of IL-18, IFN-gamma and Fas-L was analyzed by RT-PCR. We further performed IL-18 immunostaining of normal and tumor specimens. The results were correlated with tumor dissemination and clinical outcome. We report the synthesis of IL-18 in human normal colon, mainly by epithelial cells of the mucosa. Out of the 6 tumor cell lines, 4 expressed IL 18 transcripts, but neither ICE mRNA nor secreted forms of IL-18 were detected. We observed decreased or abolished synthesis of IL-18 in colon adenocarcinomas, as compared with normal mucosa. Thus, half of the colon-cancer tissues (7/14 cases) expressed neither IFN-gamma nor Fas-L. This feature was correlated with the existence of distant metastases (Fischer's exact test, p = 0.02) and an unfavorable outcome. These findings suggest that production of IL-18 in human colon may play a role in homeostasis and in tumor immune surveillance, by enhancing IFN-gamma production and Fas-L-dependent cytotoxicity of immune cells. PMID- 10371357 TI - Fungal infections after bone marrow transplant. AB - With improved control of cytomegalovirus infection, invasive fungal infections have become the leading cause of infectious mortality after bone marrow transplantation (BMT). A number of changes in transplant practices have led to changes in patterns of fungal infections: neutropenic episodes have been shortened through the use of hematopoietic growth factors and peripheral blood as a source of stem cells. More potent immunosuppressive regimens, including T-cell depletion techniques, have encouraged the use of alternate donor sources with greater numbers of transplant recipients experiencing more prolonged and more profound immunodeficiency following engraftment. The advent of new antifungal agents has led to a decline in Candida infections, but has encouraged the emergence of other less susceptible fungal pathogens. The development of molecular techniques to distinguish different fungal strains has led to identification of nosocomial transmission as an unexpected means for the spread of fungal infections in BMT units. These shifts in fungal infection patterns emphasize the need for infection control monitoring. The development of more accurate diagnostic tools and the incorporation of new antifungal agents into practice are needed to further improve outcomes. PMID- 10371358 TI - Quantity and quality of engrafting cells in cord blood and autologous mobilized peripheral blood. AB - Cord blood (CB) and autologous mobilized peripheral blood stem/progenitor cells (PBSC) are now used widely for clinical transplantation. We characterized the short-term (<8 weeks) and long-term (>8 weeks) engraftment in NOD/SCID mice resulting from transplanted CD34+ cells from these two sources. We also quantified the frequency of long-term engrafting cells, and the average proliferative capacity of individual engrafting cells by a competitive repopulation assay with binomial variance-covariance modeling. We found that 0.5 million CD34+ CB cells were able to generate sustained, high-level, multilineage human hematopoiesis, whereas a sixfold higher number of CD34+ PBSC (3 million) from cancer patients undergoing chemotherapy generated comparable short-term, but much lower sustained multilineage human hematopoiesis after transplantation. In comparison to CD34+ cells from PBSC from cancer patients, long-term engrafting cells were approximately eightfold enriched in CB CD34+ cells, and each CB long term engrafting cell had an approximately 15-fold higher multilineage proliferative capacity. Thus, the number and function of transplantable hematopoietic cells were remarkably different between these two sources of stem/progenitor cells. PMID- 10371359 TI - HLA-C disparity between patients and unrelated donors matched for HLA-A, -B, and DRB1 alleles: impact of serological vs. DNA typing for HLA-A and -B loci. AB - High incidences of graft failure, graft-vs.-host disease (GVHD), and serious infections following unrelated donor (URD) marrow transplantation, despite apparent human leukocyte antigen (HLA) identity, may reflect the presence of molecular disparities, including those for HLA-C alleles between the patient and the URD. The level of these disparities could be significant, because as many as 42 alleles are currently known for HLA-C locus. We studied 84 patients and 251 potential URDs to evaluate 1) the extent of HLA-C disparity between the patient and the URD identified by serology for HLA-A and -B and by DNA typing for -DRB1 and 2) the level of HLA-C disparity between patients and URDs matched by high resolution DNA typing for HLA-A, -B, and -DRB1. The DNA typing was performed at the Memorial Sloan Kettering Cancer Center, and the serotyping was provided by the registries. Of 251 URDs matched by HLA-A and -B serology and -DRB1 (sA_sB_dnaDRB1 ); 94, 75, and 82 were 6/6, 5/6, and 4/6 matches, respectively. Of 94 sA_sB_dnaDRB1 6/6 URDs, 51 (54.3%) were matched for both HLA-C alleles. In contrast, 31 (41.3%) 5/6 (p=0.12) and 15 (18.3%) 4/6 (p < 0.01) sA_sB_dnaDRB1 URDs were matched for both HLA-C alleles. Following DNA typing for HLA-A and -B, 52 (55.3%) of 94 6/6, 30 (40%) of 75 5/6, and 25 (30.5%) of 82 4/6 sA_sB_dnaDRB1 URDs remained 6/6, 5/6, and 4/6 matches at the DNA level (dnaA_B_DRB1). HLA-C disparities continued to exist in the dnaA_B_DRB1 URD group. Of 54 dnaA_B_DRB1 6/6 URDs, 41 (75.9%) were matched for both HLA-C alleles. Only 45.3% of the 5/6 (p=0.01) and 22.2% of the 4/6 (p < 0.01) dnaA_B_DRB1 URDs were matched for both HLA-C alleles. In the 6/6 category, the frequency of HLA-C matching improved (75.9 vs. 54.3%; p=0.01) following DNA matching for HLA-A and -B. In comparison to mismatching for HLA-B locus, mismatching for either HLA-DRB1 or -A resulted in a lower odds ratio for HLA-C disparity. The presence of a common haplotype in the sA_sB_dnaDRBl (p=0.06) URD category improved the level of HLA-C matching. We identified alleles that are associated with high (B*1501, B*4402, B*5101, DRB1*0101, A*0201, A*1101, A*2301, and A*3201) or low (B*0702, B*0801, B*1302, B*3502, DRB1*0301, DRB1*1104, A*0101, A*3001, and A*6801) probability of HLA-C disparity. Overall, sA_sB_dnaDRB1 as well as dnaA_B_DRB1 matched URDs for non Caucasian patients were more likely to have HLA-C disparity in comparison to the matched URDs of Caucasian patients. However, a high incidence of HLA-C disparities was identified even in the URDs for Caucasian patients. Whether the disparities demonstrated by this study contribute to the higher immunological complications noted following URD bone marrow transplantation is unclear. Outcome analysis and studies aimed at understanding the functional role of HLA-C may provide an answer. PMID- 10371360 TI - A novel four-drug ablative regimen with hemopoietic stem cell rescue for patients with breast cancer: a phase II study. AB - High-dose chemotherapy and autologous hemopoietic stem cell transplantation (HSCT) may provide durable progression-free survival in some patients with stage IV breast cancer (S4Brca). We have studied a new four-drug intensive preparative regimen with HSCT in a group of 158 women with S4Brca to define the risk and potential benefit of this regimen in this patient population. From May 1988 through May 1997, 158 women with S4Brca at a single center were treated with cisplatin, etoposide, thiotepa, and cyclophosphamide (PETCy) plus autologous HSCT Eligible patients were also treated with posttransplant involved-field radiation therapy. Patients with estrogen-receptor positive tumors not previously treated with tamoxifen also received this therapy for 5 years following transplantation. All patients experienced significant toxicity requiring blood-product support and parenteral nutrition. Eighteen patients (11%) died of regimen-related toxic events. With a median follow-up of 540 days for surviving patients, a retrospective Kaplan-Meier analysis projects an overall survival of 38+/-8.5% (95% CI) at 890 days with a maximum follow-up of 8.8 years. For 52 patients in sensitive relapse, the median event-free survival time is 767 days, with 46.2+/ 15.3% (95% CI) predicted to be alive at 884 days with a maximal follow-up of almost 9 years. Nearly one-half of patients in this study with S4Brca in sensitive relapse have experienced durable remissions following PETCy ablation and HSCT. Although toxicity is significant, the PETCy regimen produces a favorable balance between efficacy (event-free survival) and treatment failure (relapse + regimen-related toxic death) compared with published results. These data suggest that within the high-dose range for preparative therapy, a steep dose-response may exist for breast cancer. Trials comparing the dose intensity of preparative regimens are warranted. PMID- 10371361 TI - Relationship of tacrolimus whole blood levels to efficacy and safety outcomes after unrelated donor marrow transplantation. AB - Tacrolimus has proved effective for preventing acute graft-vs.-host disease (GVHD) after unrelated donor marrow transplantation, but the therapeutic window is apparently narrow. Therapeutic drug monitoring could potentially be used to guide dose modifications, but the optimal target range of tacrolimus blood concentrations is unknown. We determined whether acute GVHD and renal dysfunction correlated with tacrolimus whole blood levels as measured by the IMx assay. Data were analyzed for 97 adults treated in a multicenter trial of tacrolimus and methotrexate or methylprednisolone as GVHD prophylaxis after unrelated donor marrow transplantation. The rate of grades II-IV GVHD was 49%; 81% of patients had a doubling of the serum creatinine; and 61% had a serum creatinine <2 mg/dL. The initial tacrolimus target range for the clinical trial was 10-60 ng/mL. Tacrolimus blood levels were averaged over a 14-day period, and Cox models were used with averaged blood levels as a time-dependent covariate. No significant change was observed in the risks of acute GVHD, doubling of creatinine, need for dialysis, or death over a tacrolimus blood level range of 5-40 ng/mL, but there was a direct correlation between risk of developing a creatinine <2 mg/dL and increasing tacrolimus blood levels (4.7% increased risk for each 1 ng/mL increase in blood concentration, p < 0.001). When the tacrolimus level exceeded 20 ng/mL, there was a 2.2-fold increase in the rate of renal toxicity (p < 0.001), a trend for an increase in mortality (relative risk 3.9, p=0.08), and no impact on the risk of GVHD. This analysis supports 10-20 ng/mL as the therapeutic range of tacrolimus whole blood steady state or trough levels for unrelated donor marrow transplantation. PMID- 10371362 TI - Atenolol and fetal growth in pregnancies complicated by hypertension. AB - Atenolol use may be associated with growth retardation when given in pregnancy, although the relationship to trimester of initiation, duration of treatment, and its use as monotherapy is still uncertain. To compare the obstetric and fetal outcome between women receiving atenolol (as monotherapy) and other antihypertensive drug monotherapies, and also to investigate the effect of duration of treatment on fetal growth, we performed a retrospective cohort study of 312 pregnancies in 223 women attending an Antenatal Hypertension Clinic. Atenolol (as monotherapy) was given in 78 pregnancies (25.0%), other types of antihypertensive drugs as monotherapy were given in 53 pregnancies (17.0%), and multiple drug combinations were given in 90 pregnancies (28.8%). In 91 pregnancies (29.2%) no antihypertensive drugs were given. Atenolol was found to be associated with lower birth weight and ponderal index values, with a trend toward a higher prevalence of preterm (<37 weeks) delivery and small-for gestational-age babies when compared to other antihypertensive drugs as monotherapy, or to no treatment. The adverse effect of atenolol was more pronounced in women receiving the drug earlier in their pregnancy, and continuing the drug for a longer duration. In conclusion, atenolol should be avoided in the early stages of pregnancy and given with caution at the later stages, as it is associated with fetal growth retardation, which is related to duration of treatment. PMID- 10371363 TI - A clinical trial to improve high blood pressure care in young urban black men: recruitment, follow-up, and outcomes. AB - This randomized trial recruited and followed underserved, inner-city, hypertensive (HTN), young black men and investigated whether a nurse-community health worker team in combination with usual medical care (SI) increased entry into care and reduced high blood pressure (HBP), in comparison to usual medical care (UC) alone. Emergency department records, advertising, and BP screenings identified potential participants with HBP. Telephone calls and personal contacts tracked enrollees. Of 1391 potential participants, 803 (58%) responded to an invitation to be screened and scheduled a visit. Of these, 528 (66%) kept an appointment, 207 (35%) were BP eligible, and 204 (99%) consented to enroll. At 12 months 91% of men were accounted for and 85.8% (adjusted for death, in jail, or moved away) were seen. Mean BP changed from 153(16)/98(10) to 152(19)/94(11) mm Hg in the SI group and 151(18)/98(11) to 147(21)/92(14) mm Hg in the UC group (P = NS). High rates of participation are attainable in this population; however, culturally acceptable ways of delivering HBP care are needed. PMID- 10371364 TI - The renin-angiotensin system and blood pressure: differences between blacks and whites. AB - Although blacks have lower plasma renin activity compared to whites, the corresponding differences in serum angiotensin converting enzyme (ACE) levels have not been well studied. Furthermore, few studies have examined the relationship of renin activity and ACE levels to blood pressure (BP) in blacks. We addressed these questions in a cross-sectional study conducted in 110 blacks and 183 whites who were not on antihypertensive medications. Three BP readings were obtained during a clinic visit. Plasma renin activity was assayed by radioimmunoassay and serum ACE levels were measured by spectrophotometry. Mean systolic and diastolic BP were 122.6 and 77.9 mm Hg in the blacks, and 123.4 and 77.9 mm Hg in the whites, respectively. Plasma renin activity was significantly lower in the blacks compared to the whites (0.92 v 1.26 ng/mL/h, respectively, P < .05), but ACE levels were similar in both groups (28.8 v 29.6 U/L, respectively). Renin activity was significantly and inversely associated with systolic and diastolic BP in both the blacks and the whites. ACE levels, however, were inversely associated with BP in the blacks but positively associated with BP in the whites (P = .02 for interaction on diastolic BP), even after adjustment for age, gender, body mass index (BMI), alcohol consumption, and heart rate. The corresponding interaction between ACE level and race on systolic BP was of borderline significance (P = .06). These results suggest that levels of ACE are similar in blacks and whites but their association with BP is possibly reflecting underlying ethnic differences in regulation of BP. PMID- 10371365 TI - Sustained antihypertensive actions of a dual angiotensin-converting enzyme neutral endopeptidase inhibitor, sampatrilat, in black hypertensive subjects. AB - Our objective was to evaluate the safety and antihypertensive efficacy of sampatrilat, a novel dual inhibitor of both angiotensin-converting enzyme (ACE) and neutral endopeptidase (NEP), in subjects poorly responsive to ACE inhibitor monotherapy. The ability of sampatrilat (50 to 100 mg daily) (n = 28) to lower blood pressure was compared with that of the ACE inhibitor lisinopril (10 to 20 mg daily) (n = 30) using a double-blind, randomized, parallel group study design over a 56-day treatment period in black hypertensives. Changes in systolic (SBP) and diastolic (DBP) blood pressure were determined using repeated ambulatory blood pressure (ABP) monitoring. Both sampatrilat and lisinopril decreased plasma ACE concentrations after 28 and 56 days. The decrease in plasma ACE concentrations (U/L) was greater after lisinopril (-9.33 +/- 0.52) as compared with sampatrilat (-6.31 +/- 0.70) (P = .0001) therapy. Lisinopril, but not sampatrilat, increased plasma renin activity. Lisinopril produced a transient decrease in mean 24-h ABP (mm Hg) at 28 days (SBP = -9.0 +/- 2.3, DBP = -5.7 +/- 1.3; P < .01), which returned to pretreatment values by 56 days of therapy. Alternatively, sampatrilat produced a sustained decrease in mean ABP over the 56 day treatment period (day 28: SBP = -7.3 +/- 1.8, DBP = -5.2 +/- 1.2; P < .01: day 56: SBP = -7.8 +/- 1.5; DBP = -5.2 +/- 0.95; P < 0.01) with a greater treatment effect on DBP than that of lisinopril at day 56 (P = .05). Treatment emergent adverse events were noted to be similar between both treatment groups. We conclude that the antihypertensive actions of ACE/NEP inhibitor monotherapy in black subjects offers a novel therapeutic approach to patients otherwise resistant to the sustained antihypertensive actions of ACE inhibitor monotherapy. PMID- 10371366 TI - Altered sympathetic and vagal modulations of the cardiovascular system in patients with pheochromocytoma: their relations to orthostatic hypotension. AB - To examine sympathetic and vagal cardiovascular regulatory mechanisms in the pathogenesis of orthostatic hypotension in pheochromocytoma, we continuously monitored blood pressure (Finapres) and RR interval (electrocardiogram) in supine and standing positions in 12 patients with pheochromocytoma, 43 patients with essential hypertension, and 30 normotensive subjects. Mayer wave power spectrum of systolic blood pressure variability (approximately 0.1 Hz) and respiratory power spectrum of the RR interval variability (approximately 0.25 Hz) were taken as measures of sympathetic vascular and cardiac vagal modulations, respectively. Systolic blood pressure decreased more upon standing in pheochromocytoma patients (-21 +/- 7 mm Hg) than in normotensive subjects (-5 +/- 2 mm Hg) or essential hypertensive patients (-3 +/- 2 mm Hg) (P < .005 for both), whereas heart rate tended to increase most in the pheochromocytoma group. Postural reduction in systolic blood pressure was highly correlated with postural increase in heart rate (reciprocal change in RR interval) in the pheochromocytoma group (r = 0.716, P < .01) suggesting that baroreflex is well functioning in those patients. The Mayer wave power spectrum in recumbency was extremely depressed in pheochromocytoma patients (1.1 +/- 0.2 mm Hg2) compared with normotensives (4.5 +/- 0.8 mm Hg2) or essential hypertensives (5.6 +/- 0.6 mm Hg2) (P < .001 for both). This parameter increased significantly with standing in all groups but remained lower in patients with pheochromocytoma (5.1 +/- 1.0 mm Hg2) than in normotensives (7.1 +/- 0.9 mm Hg2, P = NS), whereas essential hypertensive patients demonstrated far greater value (19.2 +/- 3.8, P < .01 for both). The respiratory power spectrum of the RR interval in recumbency of pheochromocytoma patients (189 +/- 54 msec2) was less than in normotensive subjects (714 +/- 100 msec2, P < .001) but did not differ from that in patients with essential hypertension (214 +/- 41 msec2). The respiratory power spectrum of the RR interval upon standing was markedly suppressed in pheochromocytoma patients (36.9 +/- 16.7 msec2) compared with normotensive subjects (129.5 +/- 23.6 msec2) or essential hypertensive patients (126.6 +/- 28.6 msec2) (P < .001 for both). Postural decrement in the respiratory power spectrum of the RR interval correlated positively with postural increase in heart rate (r = 0.577, P < .05) in patients with pheochromocytoma. After successful surgery (n = 9), the Mayer wave power spectrum of the systolic blood pressure and the blood pressure response to orthostasis were normalized. These data suggest that altered sympathetic vascular regulation is central to the pathogenesis of orthostatic hypotension in pheochromocytoma, whereas cardiac vagal regulation acts to compensate. PMID- 10371367 TI - Metabolic, hemodynamic, and cardiac effects of captopril in young, spontaneously hypertensive rats. AB - Spontaneously hypertensive rats (SHR) demonstrate elevated blood pressure, cardiac hypertrophy, glucose intolerance, and insulin resistance compared with age-matched Wistar-Kyoto rats (WKY). We investigated concurrent effects of captopril on blood pressure, cardiac mass, myocardial enzyme activities, glucose tolerance, and insulin action in young male SHR. At 10 weeks of age, SHR were randomized into two groups, one receiving distilled water, the other a captopril solution (50 mg/kg body weight/day). We also examined age-matched WKY receiving distilled water. Blood pressure was measured by tail-cuff during the 4-week treatment period and oral glucose tolerance was tested at the end of treatment. Hearts were weighed and ventricular tissue was assayed for activities of 3 hydroxyacyl-CoA dehydrogenase, citrate synthase, and hexokinase. Growth rates were similar between captopril-treated and control SHR, but less than those of WKY. Captopril reduced blood pressure (134 +/- 8 v 177 +/- 8 mm Hg, P < .05) and left ventricular mass (-18%, P < .05) in SHR. Cardiac enzyme activities also changed with captopril treatment, reflecting an increased capacity for beta oxidation of fatty acids and reduced potential for glucose phosphorylation in the left ventricle of SHR. Serum concentrations of glucose, insulin, and free fatty acids after a brief fast and in response to oral glucose were not different after captopril treatment, suggesting no improvement in insulin action or glucose tolerance. In summary, treatment of young male SHR with captopril reduces blood pressure and cardiac mass, and promotes a small but significant increase in cardiac capacity for oxidation of fatty acids and reduction of glucose phosphorylation. In contrast, metabolic effects of captopril on oral glucose tolerance and insulin action were not evident. PMID- 10371368 TI - Sustained contraction to angiotensin II and impaired Ca2+-sequestration in the smooth muscle of stroke-prone spontaneously hypertensive rats. AB - Contractile response to angiotensin II (AngII) of vascular smooth muscle was compared between 12-week-old, stroke-prone spontaneously hypertensive rats (SHRSP) and Wistar-Kyoto rats (WKY). AngII induced phasic and tonic contraction in denuded aortic ring preparation. The phasic contraction was concentration dependent (AngII 10(-9) to 10(-6) mol/L) and similar in both strains. However, the relaxation after phasic contraction was significantly attenuated in SHRSP compared with that in WKY. To examine the recovery of contractile responses to the repeated stimulation, AngII was applied three times at 20- and 60-min intervals. The first maximal contraction was similar in both strains, but the response to the second stimulation was significantly reduced in SHRSP, compared with all three responses in WKY. These results suggested that Ca2+-sequestration into the Ca2+ store is delayed in SHRSP. Cyclopiazonic acid (10(-5) mol/L), an intracellular Ca2+-pump inhibitor, decreased spontaneous relaxation and increased the sustained contraction in WKY, whereas it did not affect the contraction in SHRSP. Insulin, which modulates tonic contraction by facilitating Ca2+-extrusion, was applied at peak contraction by AngII. It enhanced relaxation after phasic contraction in a concentration-dependent manner in SHRSP, but it did not affect the relaxation in WKY. These results suggest that increased sustained contraction observed in SHRSP reflects at least partly the impaired Ca2+-pump activity leading to hypertension. PMID- 10371369 TI - Alteration of muscle fiber composition linking to insulin resistance and hypertension in fructose-fed rats. AB - The aim of this study was to examine the role of muscle fiber composition in insulin resistance and the effect of a calcium channel antagonist on insulin sensitivity in fructose-induced insulin resistant and hypertensive rats. Six-week old male Sprague-Dawley rats were fed either normal rat chow (control) or fructose-rich diet (FFR). For the last 2 weeks of a 6-week period of either diet, the rats were treated, by gavage, with gum arabic solution (control or FFR) or a dihydropyridine calcium channel antagonist, benidipine hydrochloride (3 mg/kg/day: FFR + Ca), then the euglycemic hyperinsulinemic glucose clamp technique was performed to evaluate insulin sensitivity. Blood pressure was measured weekly for 6 weeks. At the end of the glucose clamp, the soleus muscle was dissected out for determination of muscle fiber composition by ATPase methods. Blood pressure was elevated at 2 weeks after the start of fructose-rich chow feeding and persisted thereafter throughout the study. Blood pressure at the glucose clamp in the FFR was significantly higher than that in the control group (142 +/- 2 v 155 +/- 2 mm Hg, P < .01) and the calcium antagonist significantly lowered blood pressure of FFR (136 +/- 6 mm Hg for FFR +/- Ca, P < .05). The average rate of glucose infusion during glucose clamp, as a measure of insulin sensitivity (M value), was significantly lower in the FFR than in the control (15.4 +/- 0.4 v 10.9 +/- 0.6 mg/kg/min, P < .01). The calcium channel antagonist partially improved the M value compared to that of FFR (13.4 +/- 0.7 mg/kg/min in FFR +/- Ca, P < .01 compared to FFR, P < .05 compared to control). The composite ratio of type I fiber in soleus muscle was significantly decreased in FFR compared to control (81.7 +/- 1.5% v 75.0 +/- 1.7%, P < .01), and the composite ratio of type I fiber in rats treated with the calcium channel antagonist (FFR +/ Ca) recovered to the control level (79.9 +/- 1.1%, P < .05 compared to FFR). The M value was significantly correlated with the compositions of type I and type II fibers (for type I fibers, r = 0.80, P < .01; for type II fibers, r = -0.81, P < .01). These results suggest that fiber composition of skeletal muscle links insulin resistance and that a calcium channel antagonist may modulate muscle fiber composition in hypertensive animal model, fructose-fed rats. PMID- 10371370 TI - Genetic susceptibility of the donor kidney contributes to the development of renal damage after syngeneic transplantation. AB - Solitary kidneys, especially in rats, appear vulnerable to develop functional and structural damage. However, differences in susceptibility exist between strains. It is not clear whether this is intrinsic to the kidney or due to environmental factors. Therefore, the aim of the present study was to investigate possible differences in genetic susceptibility for renal damage. By transplanting different rat donor kidneys into a normotensive, histocompatible recipient, the kidneys were exposed to the same blood pressure profiles, metabolic and hormonal environment. Kidneys from young adult hypertensive fawn-hooded (FHH) rats, a strain showing early onset renal damage, normotensive, renal damage-resistant August x Copenhagen-Irish (ACI), and (ACI x FHH) F1 donors were transplanted into male F1 recipients. The native kidneys of the recipients were removed 1 week after transplantation. The results were mutually compared and to their unilaterally nephrectomized littermates. Systolic blood pressure (SBP) and albuminuria (UaV) were determined at the time of transplantation and at 8 and 16 weeks. The histomorphologic analysis included the incidence of focal glomerulosclerosis (FGS), and determination of chronic transplant dysfunction according to the BANFF criteria. A negative impact of the transplantation technique in this syngeneic situation could not be detected as F1 transplants did not differ functionally and morphologically from their UNx controls. Transplanting an ACI kidney did not result in significant changes of SBP, UaV, and incidence of FGS compared to F1 transplants and ACI-UNx. In contrast, FHH kidneys did show a progressive increase of UaV and glomerulosclerosis and a significantly higher BANFF score, whereas the SBP did not differ from F1 transplants. The moderate hypertension seen in FHH did not travel with the kidney. Compared to the FHH-UNx rats, transplantation of a FHH kidney did significantly attenuate the increase of UaV and FGS. The susceptibility of the donor kidney appears to be an important factor in the development of chronic renal damage. This may play a role in the long-term functional changes seen after clinical renal transplantation. PMID- 10371371 TI - Effects of digoxin-specific antibody Fab fragment (Digibind) on blood pressure and renal water-sodium metabolism in 5/6 reduced renal mass hypertensive rats. AB - The importance of increased endogenous digitalis-like factor (EDLF) in volume expanded hypertension has been generally agreed. To further clarify the role of EDLF on the development of hypertension and renal water-sodium handling in 5/6 reduced renal mass hypertensive rats (RRM), we studied the effects of acute administration of digoxin-specific antibody Fab fragment (Digibind) in the early phase and the chronic phase of hypertension in RRM. RRM and sham-operated rats were given 1% saline for 1 or 4 weeks. RRM were injected Digibind (60 mg/kg) or vehicle (0.9% saline) intravenously in the first or fourth week under thiobutabarbital anesthesia. All sham-operated rats were administered Digibind under the same condition. Digibind altered neither blood pressure, heart rate, urine volume, nor urinary sodium excretion in sham-operated rats. However, Digibind produced a gradual but significant decline in mean arterial pressure to the level slightly above that in sham-operated rats from 153 +/- 5 to 131 +/- 5 mm Hg in the first week and from 181 +/- 6 to 129 +/- 4 mm Hg in the fourth week without any significant change in heart rate. The decrease in mean arterial pressure at 160 min after Digibind administration in the fourth week (-48 +/- 5 mm Hg) was greater than that in the first week (-22 +/- 4 mm Hg). No differences were observed in urine volume, urinary sodium excretion, or plasma norepinephrine concentration between Digibind and vehicle-treated RRM in either week. These data suggest that EDLF would contribute to both the early and chronic phase in the development of hypertension in RRM. PMID- 10371372 TI - Effects on regional renal blood flow when unclipping a two-kidney, one-clip hypertensive Wistar rat during renal nerve stimulation. AB - Blood pressure (BP) is rapidly normalized when removing the obstruction from the renal artery of a two-kidney, one-clip renovascular hypertensive rat (unclipping). This study tested whether efferent renal nerve stimulation (ERNS) of the unclipped kidney affects this drop in BP or the associated changes in diuresis-natriuresis and regional renal blood flow. Three groups of anesthetized renovascular hypertensive Wistar rats were studied: 1) W(C) (time control); 2) W(UC) (unclipped after 30 min); and 3) W(UC+NS) (unclipped after 30 min, with ERNS at 5 Hz for 2 h). Renal excretion and regional hemodynamics (laser Doppler) were monitored in the unclipped kidney. Medullary and cortical blood perfusion increased by 84% and 95%, respectively, in W(UC) 30 min after unclipping (P < .001) but only with 8% and 9%, respectively, in W(UC+NS) (P = NS). Unclipping induced a marked increase in diuresis-natriuresis that was largely unaffected by ERNS. In W(UC) and W(UC+NS) BP returned to normotensive levels within 4 h. However, during the first 30 min, average BP decreased significantly less in W(UC+NS) (9%, 20 mm Hg) than in W(UC) (16%, 35 mm Hg) (P < .05). ERNS at 5 Hz effectively prevented the increase in medullary blood perfusion but did not affect the fall in blood pressure or the pressure diuretic/natriuretic response seen after unclipping. The results suggest that both the reduction in BP and the pressure-induced increase in diuresis/natriuresis seen when unclipping the 2K,1C renovascular hypertensive rat occurs largely independently of ERNS and an increase in medullary blood perfusion. PMID- 10371373 TI - Association between resting heart rate and hypertension treatment in a general population. AB - Epidemiologic studies have reported a relationship between resting heart rate (RHR) and cardiovascular mortality, particularly in hypertensive subjects. In a representative sample (n = 1175) of women and men aged 35 to 64 years, we studied the associations between RHR and hypertension. RHR was associated with sex (P < .001), socioeconomic and marital status (P < .05), physical fitness (P < .001), smoking (P < .05), hypercholesterolemia (P < .01), body mass index (P < .01), blood pressure (P < .01), triglyceride levels (P < .01), glycemia (P < .05), hematocrit (P < .001 in men, not significant in women), and white blood cell count (P < .01). Logistic regression models adjusted for the above variables were developed with RHR coded as a polytomous outcome variable (RHR < 65/min; 65 < or = RHR < 75/min; 75 < or = RHR < 85/min; RHR > or = 85/min). Subjects unaware of their hypertension had significant adjusted odds ratios for high RHR categories [75 < or = RHR < 85/min: 2.11 (1.37 to 3.23), P < .001; RHR > or = 85/min: 4.71 (2.06 to 10.78), P < .001]. People treated for hypertension had nonsignificant odds ratios whatever the RHR categories. After adjustment for numerous risk factors, elevated RHR were associated with high blood pressure in unaware hypertensive subjects. The impact of antihypertensive drugs with different RHR lowering effects remains to be studied on a population basis. PMID- 10371374 TI - The Gly460Trp variant of alpha-adducin is not associated with hypertension in white Anglo-Australians. AB - A Gly460Trp variant of the cytoskeletal protein alpha-adducin has recently been implicated in the etiology of essential hypertension (HT) in a study involving southern European whites. We attempted to replicate this finding in a well characterized, extensively studied group of 112 white Australians with essential HT, with strong family history (two HT parents), early-onset, moderate to severe disease, and of British extraction. Controls were 196 normotensive (NT) white subjects whose parents were both NT older than age 50 years. A mismatch polymerase chain reaction method involving BanII was developed for genotyping. Frequency of the Trp460 allele was 0.23 in the HT and 0.24 in the NT groups (chi2 = 0.2, P = .7). No association was observed with blood pressure, body mass index, age, plasma renin, angiotensinogen, angiotensin converting enzyme, cholesterol, triglycerides, or HDL or LDL cholesterol. Our results therefore provide no support for a role for the alpha-adducin variant in hypertension, at least in our severely affected Anglo-white group with strong family history of HT. PMID- 10371375 TI - Deletion polymorphism of the angiotensin converting enzyme gene is associated with an increase in left ventricular mass in men with type 2 diabetes mellitus. AB - Previous studies evaluating the angiotensin converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism have revealed that expression of the DD genotype is associated with an increase in myocardial infarction, cardiomyopathy, and left ventricular (LV) mass in nondiabetic patients. In the present study, a cross-sectional analysis was performed to evaluate the potential relationship between the ACE I/D genotypes and the LV mass index in 289 non-insulin-dependent diabetes mellitus (NIDDM) subjects without known coronary artery disease. Two dimensional directed M-mode echocardiograms along with selected patient characteristics were obtained from the study population. The distribution of the I/D polymorphism was as follows: 63 were II (22%), 137 were ID (47%), and 89 were DD (31%). Univariately, the DD genotype was associated with an increase in LV mass in men but not in women. When subjected to a multiple regression model that included age, systolic blood pressure, duration of diabetes, duration of hypertension, presence of the black race, and the presence of the DD genotype, the DD genotype was independently associated with an increase in the LV mass index with a parameter estimate of 10.5 g/m2 (95% CI = 3.9, 17.0; P < .002) in the male subjects. Thus, in this NIDDM study population, male patients with the DD genotype are independently associated with an increased LV mass. PMID- 10371376 TI - Moderate dietary salt restriction increases vascular and systemic insulin resistance. AB - Our recent studies have indicated that severe salt restriction aggravates vascular insulin resistance in younger normotensive and hypertensive subjects. However, whether the extent of dietary salt restriction commonly advocated adversely affects vascular insulin resistance is unknown. To determine whether moderate dietary salt restriction might affect vascular and systemic sensitivity to insulin, we studied eight subjects after 1 week of a normal sodium diet (235 mEq/day) and 1 week of a moderate salt restriction (75 meq/day). Systemic insulin resistance as assessed by the fasting plasma glucose-to-insulin ratio was aggravated by dietary sodium restriction (normal sodium: 1.2 +/- 0.1 mmol/mIU; low sodium 0.6 +/- 0.1, P < .05). Salt restriction significantly reduced maximal insulin-mediated vasodilation (normal sodium: 51% +/- 5% of maximum nitroglycerin mediated response; low sodium: 28% +/- 6%, P < .01). In contrast, no alterations in nitroglycerin-mediated vasodilation nor phenylephrine-mediated vasoconstriction were noted. These studies demonstrate that moderate salt restriction aggravates both systemic and vascular insulin resistance. This impairment of the vasodilating effect of insulin could be a factor attenuating the blood pressure lowering effect of a low sodium diet. PMID- 10371377 TI - Na+ and Mg2+ contents in smooth muscle cells in spontaneously hypertensive rats. AB - Whereas in blood cells decreased magnesium concentrations and increased sodium concentrations in essential hypertension have often been described, only sparse data exist on cellular magnesium or sodium content and exchange in vascular smooth muscle cells. Therefore in aortic smooth muscle cells from 10 spontaneously hypertensive rats (SHR) of the Munster strain and 10 normotensive Wistar-Kyoto rats (WKY) aged 8 to 10 months, the intracellular magnesium and sodium content was measured. Electron-probe X-ray microanalysis was used to determine intracellular Mg2+ and Na+ concentrations in aortic cryosections 3 microm thick. The magnesium ion content was 0.90 +/- 0.15 g/kg dry weight in SHR versus 1.15 +/- 0.10 g/kg dry weight in WKY (means +/- SD, P < .05). Vascular smooth muscle sodium ion content was 6.66 +/- 0.39 g/kg dry weight in WKY and 12.61 +/- 0.91 g/kg dry weight in SHR (P < .01). Aortic smooth muscle cells from SHR are characterized by markedly lowered intracellular magnesium ion content and increased sodium ion concentrations in animals 8 to 10 months old, compared with normotensive cells. The results may be due to genetically determined disturbances in transmembrane magnesium and sodium ion transport. PMID- 10371378 TI - A study of topiramate pharmacokinetics and tolerability in children with epilepsy. AB - The pharmacokinetic and safety profile of topiramate as adjunctive therapy was assessed in pediatric patients with epilepsy in an open-label, 4-week, single center study. Six children from each of the following age groups were enrolled: 4 7 years, 8-11 years, and 12-17 years. Patients received topiramate 1 mg/kg/day for 1 week, with subsequent progressive weekly increases in dosage to 3, 6, and then 9 mg/kg/day or 800 mg/day, whichever was less. Topiramate oral plasma clearance (CI/F) was independent of dose, and steady-state plasma concentrations increased in proportion to dose. Weight-normalized topiramate CL/F was higher (P = 0.003) in pediatric patients receiving enzyme-inducing concomitant antiepileptic drugs (AEDs) (mean = 70.1 ml/minute/70 kg) than in those not receiving enzyme-inducing AEDs (mean = 33.1 mL/ minute/kg). Topiramate CL/F in children was approximately 50% greater than that observed in adults regardless of the type of concomitant AED therapy. Thus steady-state plasma topiramate concentrations for the same mg/kg dose will be approximately 33% lower in pediatric patients than in adult patients. The most frequently reported treatment emergent adverse events considered related to topiramate therapy included anorexia, fatigue, and nervousness, and no patient discontinued therapy. This study indicates that, in children 4-17 years of age, topiramate has linear pharmacokinetics, 50% higher clearance than in adults, and is generally well tolerated. PMID- 10371379 TI - Epilepsy surgery in children with pervasive developmental disorder. AB - Pervasive developmental disorder (PDD) is occasionally associated with medically intractable complex partial seizures. The outcome of PDD was explored in three males and two females who underwent epilepsy surgery at 32 months to 8 years of age (mean = 4 years) after onset of epilepsy at 1 week to 21 months of age (mean = 11 months). Four children had temporal lobe resections (three right, one left; two for focal cortical dysplasia, and two for tumors), and one had a right temporoparieto-occipital resection (for focal cortical dysplasia). Each child underwent repeated evaluations by a pediatric neuropsychologist and psychiatrist. Fourteen to 47 months (mean = 23 months) after operation, one child with persistent seizures had moderate developmental and behavioral improvement, three children (two seizure free, one with rare staring spells) had mild developmental and behavioral improvement, and the remaining child (seizure free) experienced a worsening of her PDD. The four children with mild-to-moderate improvement in postoperative cognitive and behavioral development still demonstrated persistent delay. Cognitive gains were confirmed by neuropsychologic testing in the oldest patient but were not reflected in test results from the three younger children, who had more modest improvement. The child with worsening of her PDD had cognitive and emotional deterioration to babbling, echolalia, aggressiveness, decreased social interaction, and increased mouthing of objects beginning several months postoperatively. These results suggest that families should be counseled that PDD symptoms in children with focal epileptogenic lesions may or may not improve after epilepsy surgery, even if the surgery is successful with respect to seizure control. PMID- 10371380 TI - Benign epileptic discharges in patients with lesional partial epilepsies. AB - Case reports of four patients with therapy-resistant lesional partial epilepsies and additional foci of benign epileptic discharges of childhood, in addition to the usual electroencephalogram (EEG) changes, are presented. A family history of epileptic or febrile seizures in childhood was reported in all four patients. A distant relative of one patient, not manifesting seizures, demonstrated rolandic spikes on EEG. An abnormal pregnancy (polyhydramnion, premature pains, induced labor because of an abnormal CTG , placenta insufficiency) was reported in one patient, risk factors during birth (birth 14 days after term, placenta insufficiency) were reported in one, and bacterial meningitis at 4 weeks of age was reported in one. All patients manifested a retarded, partly severe, unfavorable infantile psychomotor development. An early seizure onset was observed in all patients (in three patients during the first year of life and in one patient during the second year). A hemifacial seizure symptomatology was seen, in addition to other symptoms, in two patients, possibly indicating the seizure pattern indicative of benign partial seizures; seizures occurred exclusively in sleep in one patient. The benign focus was never located in the lesional area. It was recorded over the same hemisphere in two patients and over the other hemisphere in the other two. PMID- 10371381 TI - Charcot-Marie-Tooth disease type 1A duplication by PCR analysis. AB - By generating new junctional fragments from the recombinant Charcot-Marie-Tooth (CMT) 1A-REPs in CMT1A patients, a 3.2-kb recombination hot spot is observed in three quarters of CMT1A patients. By a polymerase chain reaction (PCR) method the authors analyzed eight patients CMT1A duplication, confirmed by Southern blot, to detect a recombination hot spot. Four patients had a novel 3.2-kb junctional fragment by PCR analysis. These four patients with a novel 3.2-kb junctional fragment had an abnormal 1789-bp fragment in addition to 1986-bp fragment after NsiI digestion (type 1). One patient who demonstrated no novel 3.2-kb junctional fragment had an abnormal 336-bp fragment in addition to 265 bp (type 2). Three patients with CMT1A duplication were not diagnosed as having CMT1A on the basis of PCR analysis. The PCR-based DNA test is valuable for screening to detect CMT1A duplication. PMID- 10371382 TI - Pediatric status epilepticus: a perspective from Saudi Arabia. AB - The purpose of this study was to assess risk factors and management of status epilepticus and non-status epilepticus seizures at a community hospital in Saudi Arabia. The research design was a prevalence study of a convenience sample of pediatric seizure episodes admitted to a 350-bed hospital from 1992 to 1997. The mean age at presentation was 2 years, 10 months, 43% of patients had no history of seizures, and 17% were transferred from other hospitals. Fifty-nine (28%) of 212 seizure episodes were status epilepticus (SE). These SE episodes were significantly more likely than non-SE episodes to be associated with a history of seizures, prior antiepileptic drug (AED) therapy, the presence of an acute etiology, and prolonged duration of seizures before hospitalization. SE episodes were also significantly more likely than non-SE episodes to receive an inappropriate AED, to require intensive care unit admission, to suffer morbidity, and to have SE recurrence at follow-up; however, the difference in mortality was not significant. In conclusion, children with SE were more likely than those with non-SE seizures to have a history of seizures and acute brain insults, prolonged seizure duration before hospitalization, and less optimal management and outcomes. Management of SE in this referral population can be improved by more rapid access to appropriate medical care. PMID- 10371383 TI - Periventricular leukomalacia: relation to gestational age and axonal injury. AB - Eighty-five infants ranging from 22 to 41 weeks gestation were diagnosed as having periventricular leukomalacia (PVL) using traditional neuropathologic methods. The lesions were also studied by immunocytochemistry for beta-amyloid precursor protein (beta-APP), a glycoprotein that has been observed in PVL. Using this technique, the distribution of white matter tissue necrosis was defined as focal, widespread, and diffuse. The type of PVL correlated with the gestational age at birth. The youngest infants tended to demonstrate widespread necrosis, and the oldest infants exhibited more focal necrosis. beta-APP immunopositivity was present in the axons around the foci of white matter necrosis in 76% of the patients and in the neurons of the adjacent cortex in 66% of the patients. In age matched control patients, there was no beta-APP reactivity in the cerebral white matter or the cortex. In most patients the distribution of beta-APP-positive axons proved to be a useful marker for demonstrating the type of PVL; however, the relationship of beta-APP to the pathogenesis of PVL requires further study. PMID- 10371384 TI - MRI and MRS in HMG-CoA lyase deficiency. AB - 3-Hydroxy-3-Methylglutaryl coenzyme A lyase (HMG-CoA) deficiency is a rare inborn error of leucine catabolism. The disease is characterized by recurrent episodes of metabolic acidosis, hyperammonemia without ketosis, hypoglycemia, lethargy, hepatomegaly, and seizures. This study has evaluated the magnetic resonance imaging (MRI) and magnetic resonance spectroscopy (MRS) findings of three patients with HMG-CoA deficiency. The common findings on all of the MRI scans were multiple, coalescent, marked lesions in periventricular white matter and arcuate fibers, most prominently in frontal or periatrial regions that were superimposed on diffuse, slightly hyperintense subcortical white matter signal. Involvement of the caudate nucleus and the dentate nucleus were observed in the reported patients. MRS studies by both STEAM and PRESS spectra of all patients revealed a decrease in N-acetylaspartate and elevation in both myoinositol and choline. A pathologic peak at 1.33 ppm, which is compatible with lactate, and a particular peak at 2.42 ppm in all patients were also found. The combination of both MRI and MRS findings could be considered as being specific in patients with HMG-CoA lyase deficiency. PMID- 10371385 TI - Gabapentin for self-injurious behavior in Lesch-Nyhan syndrome. AB - Self-injurious behavior is a common clinical problem in children with Lesch-Nyhan syndrome, an X-linked disorder of purine metabolism. This behavior is not observed in other conditions associated with increased serum concentrations of uric acid, hypoxanthine, and xanthine. Various neurotransmitters appear to play a pivotal role in self-injurious behavior. The authors present a patient with Lesch Nyhan syndrome, whose self-injurious behavior was effectively treated with gabapentin, and discuss possible mechanisms of action. PMID- 10371386 TI - Multiple sclerosis presenting as a single mass lesion. AB - A 6-year-old male presented with a subacute onset of hemiplegia and a mass lesion on computed tomography scan. Pathology from a surgical biopsy revealed demyelination only. Ten months later, he had a recurrence on the opposite side. A magnetic resonance imaging scan revealed a corresponding mass lesion in the other cerebral hemisphere, allowing the diagnosis of multiple sclerosis. Consideration should be given to diagnoses other than a tumor when the clinical picture and radiologic features are atypical. PMID- 10371387 TI - Syntelencephaly associated with connected transhemispheric cleft of focal cortical dysplasia. AB - The authors report a female with syntelencephaly associated with a connected transhemispheric cleft of focal cortical dysplasia. Syntelencephaly is a rare anomaly characterized by fusion of the hemispheres in the posterior frontal and parietal regions and is considered a new variant of holoprosencephaly. Cranial magnetic resonance imaging of the patient revealed syntelencephaly associated with bilateral fused clefts of focal cortical dysplasia without the pial ependymal seam, which was regarded as an incomplete type of schizencephaly. The underlying mechanism is discussed. PMID- 10371388 TI - Electroclinical characteristics of hemimegalencephaly. AB - Presented here are two long-term follow-up patients with hemimegalencephaly. Patient 1 had Ohtahara's syndrome, which evolved into West's syndrome. Patient 2 had localization-related epilepsy, which demonstrated epilepsia partialis continua throughout the clinical course. The patients' interictal electroencephalograms revealed asymmetric suppression-burst patterns sometime during the clinical course: only during early infancy in patient 1 and until the last follow-up (at 30 years of age) in patient 2. Both patients had moderate mental and motor disturbances with persistence of seizures. Hemiplegia was progressive during early childhood. Aggravation of hemiplegia might be related to frequent seizures and persistent electroencephalographic abnormalities during early childhood. Although asymmetric suppression-burst patterns are considered characteristic electroencephalographic findings in these cases, the duration of their appearance did not have definite prognostic significance. PMID- 10371389 TI - Pediatric manifestations of Hashimoto's encephalopathy. AB - Hashimoto's encephalopathy is a steroid-responsive encephalopathy associated with elevated blood concentrations of antithyroid antibodies. The patients are usually euthyroid or mildly hypothyroid. The authors report two pediatric patients with Hashimoto's encephalopathy and review the literature. The clinical picture in adolescents, as with adults, is pleomorphic but frequently associated with seizures, confusion, and hallucinations. Alternatively, progressive cognitive decline manifested by a drop in school performance can be observed. The diagnosis of Hashimoto's thyroiditis is often overlooked at presentation and a high degree of suspicion is necessary for proper diagnosis. PMID- 10371391 TI - Joubert's syndrome and prenatal hydrocephalus. AB - Joubert's syndrome is an autosomal-recessive condition characterized by dysgenesis of the cerebellar vermis, hypotonia, developmental delay, a respiratory pattern of alternating tachypnea and apnea, and abnormal eye movements. Radiologic findings include a midline cerebellar cleft in place of the vermis and a characteristic shape of the fourth ventricle. Prenatal hydrocephalus has been proposed as a possible etiology for the cerebellar abnormalities but has not previously been described in association with this syndrome. The authors report a patient with clinical and radiographic features consistent with Joubert's syndrome who presented with congenital hydrocephalus. PMID- 10371390 TI - Acute disseminated encephalomyelitis with probable measles vaccine failure. AB - The patient is a 10-year-old male who experienced somnolence and incomplete quadriplegia after headache and vomiting, without exanthema, for 3 days. The clinical course and magnetic resonance imaging findings of the brain and spinal cord were compatible with acute disseminated encephalomyelitis. The serologic examination revealed that the patient had rubeola because titers of IgM and IgG antibody to measles virus measured by enzyme immunoassay were 0.91 and 40 (cutoff = 0.80 and 2), respectively, at 5 weeks after the onset, the IgM titer had become negative (0.56), and the IgG titer had decreased to 17.7 at 13 weeks after the onset. Because the patient had received a measles-mumps-rubella vaccine at 12 months of age, the acute disseminated encephalomyelitis was thought to be attributed to the modified measles resulting from measles vaccine failure. PMID- 10371392 TI - Phenotyping alcoholism. PMID- 10371393 TI - The heritability of alcoholism symptoms: "indicators of genetic and environmental influence in alcohol-dependent individuals" revisited. AB - There is consistent evidence from twin and adoption studies implicating genetic factors in the etiology of alcoholism, yet few studies have examined the role of genetic influences on individual symptoms of alcoholism. In a previous study of 113 male twins, Johnson et al. (1996a) identified 7 alcoholism symptoms that were more "genetic" and 14 that were more "environmental" (that is, non-genetic) in their etiology by examining symptom concordances among monozygotic and dizygotic twin pairs. The present study represents an attempt to replicate the results of this previous study and extend them by estimating the contribution of genetic factors to the variation in liability for different alcoholism symptoms. Subjects were 3356 male twin pairs from the Vietnam Era Twin Registry. Lifetime histories of alcoholism symptoms were assessed by a structured psychiatric telephone interview. The results of the previous study were not replicated. The correlations between symptom classifications as genetic and non-genetic in the present and previous study were nonsignificant and ranged from -0.27 to 0.11. However, within the present study the correlation between symptom classifications as genetic and non-genetic was statistically significant across random split-half subsamples (r = 0.59); nine alcoholism symptoms were consistently classified as genetic and six symptoms as non-genetic in their etiology. Model-fitting analyses applied to different alcoholism symptoms yielded heritability estimates ranging from 0.03 to 0.53 with broad and overlapping confidence intervals around these estimates, ranging from 0.00 to 0.65. The results of this study highlight the difficulty of identifying more or less heritable phenotypes in twin research, and suggest that it may not be possible to identify specific alcoholism symptoms that are more genetic in their etiology than others. Nevertheless, there appears to be potentially important variation in the relative magnitude of genetic influences for individual alcoholism symptoms, and exploring these differences may lead to further insights into the nosology and etiology of alcohol-related problems. PMID- 10371394 TI - Differential effects of ethanol on insulin-signaling through the insulin receptor substrate-1. AB - Insulin stimulation increases cell proliferation and energy metabolism by activating the insulin receptor substrate I (IRS-1)-signaling pathways. This downstream signaling is mediated by interactions of specific tyrosyl phosphorylated (PY) IRS-1 motifs with SH2-containing molecules such as growth factor receptor-bound protein 2 (Grb2) and Syp. Ethanol inhibits insulin stimulated tyrosyl phosphorylation of IRS-1 and DNA synthesis. This study explores the roles of the Grb2- and Syp-binding motifs of IRS-1 in relation to the inhibitory effects of ethanol on insulin-stimulated DNA synthesis, proliferating cell nuclear antigen (PCNA) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) expression, and activation of mitogen-activated protein kinase (MAPK), which is known to be essential for cell proliferation. NIH3T3 cells were stably transfected with wild-type IRS-1, or IRS-1 mutated at the Grb2 (IRS-1deltaGrb2), Syp (IRS-1deltaSyp), or Grb2 and Syp (IRS-1deltaGrb2deltaSyp)- binding sites. Cells transfected with IRS-1 had increased levels of DNA synthesis, PCNA, GAPDH, and activated MAPK. The IRS-1deltaGrb2 transfectants were highly responsive to insulin stimulation, achieving levels of GAPDH, PCNA, and activated MAPK that were higher than control. In contrast, the IRS-1deltaSyp and IRS-1deltaGrb2deltaSyp transfectants had reduced levels of DNA synthesis, PCNA, and activated MAPK. Ethanol exposure decreased insulin-stimulated DNA synthesis, PCNA, GAPDH, and activated MAPK levels in all clones, but the wild-type IRS-1 transfectants were relatively resistant, and the IRS-1deltaGrb2 transfectants were extraordinarily sensitive to these inhibitory effects of ethanol. The findings suggest that insulin-stimulated DNA synthesis and PCNA expression are mediated through the Syp-binding domain, whereas GAPDH expression and MAPK activation are modulated through both the Grb2 and Syp motifs of IRS-1. In addition, ethanol exposure may preferentially inhibit downstream signaling that requires interaction between Syp and PY-IRS-1. PMID- 10371395 TI - Ethanol elevates c-Myc levels in cultured mouse preimplantation embryos. AB - A brief exposure to ethanol accelerates the rate of early mouse embryonic development in vitro, increasing blastocyst formation, trophoblast outgrowth, and implantation rates after embryo transfer. The physiological effects of ethanol during preimplantation development are associated with rapid changes in gene expression and apparently arise from the ability of ethanol to elevate cytoplasmic free Ca2+ and alter cellular signaling pathways. The purpose of this study was to examine whether the abundance of c-Myc, a transcription factor that promotes cell proliferation and is required for blastocyst development, is upregulated in mouse blastocysts challenged with ethanol. After exposure of mouse blastocysts to 0.1% (17.5 mM) ethanol, wc determined the levels of: 1) c-Myc mRNA, using reverse transcription and the polymerase chain reaction; and 2) c-Myc protein levels, using specific monoclonal antibodies. Within 10 min of exposure to ethanol, the relative abundance of c-Myc mRNA increased 6-fold, then rapidly returned to baseline levels within 1 hr. As expected, elevation of c-Myc mRNA by ethanol was attenuated in embryos that were first treated with the intracellular Ca2+ chelator, BAPTA-AM. Western blot analysis of solubilized embryos revealed that c-Myc mRNA was translated into a single 62-kD protein that increased in intensity 30 min after treatment with ethanol. Immunocytochemical staining demonstrated that c-Myc was localized exclusively in nuclei and that staining intensity increased significantly after 10 min. Peak levels of c-Myc protein were found 30 min after ethanol exposure and persisted for at least 2 hr. The c-myc proto-oncogene seems to be an immediate early response gene for ethanol that may regulate the transcription of other genes that influence early embryogenesis and growth. PMID- 10371396 TI - Potentiation of calcium-mediated stimulation of DNA synthesis by ethanol in human and mouse fibroblasts. AB - Alcohol abuse is a risk factor for cancers of the gastrointestinal tract, and it also can precipitate psoriasis characterized by hyperproliferation of epidermal cells. Because these effects of alcohol may involve stimulation of cell growth, and ethanol (EtOH) was shown to enhance DNA synthesis in mouse fibroblasts and epidermal cells, we conducted a study to determine whether EtOH can also stimulate mitogenesis in human fibroblasts and keratinocytes. In keratinocytes, EtOH had no effects on mitogenesis after shorter (17-hr) treatments, but it partially prevented inhibition of DNA synthesis elicited by longer treatments (3 4 days) with 2 mM calcium (Ca2+), a differentiation-inducing agent. In contrast, treatment of serum-starved zinc-treated (40 microM) human skin fibroblasts with 50-60 mM EtOH for 17 hr resulted in increased DNA synthesis. EtOH-induced DNA synthesis was blocked by 1 mM EGTA, a specific Ca2+ chelator. Despite the presence of 1.8 mM Ca2+ in the cell culture medium, the addition of 1 mM extra Ca2+ (final concentration, 2.8 mM) for 17 hr induced DNA synthesis, presumably mediated by Ca2+ receptors. In eight independent human skin fibroblast lines examined, treatment with EtOH for 46 hr, but not for 17 hr, invariably enhanced the effects of Ca2+ on DNA synthesis, consistent with synergistic stimulation of cell proliferation by EtOH and Ca2+. Neomycin, a Ca2+ receptor agonist, and EtOH also exerted synergistic effects on DNA synthesis after longer (46-hr) treatments. In mouse NIH 3T3 fibroblasts, both EtOH- and Ca2+-enhanced DNA synthesis after 17-hr treatment, but they stimulated cell proliferation only in combination. The results indicate that in human fibroblasts, EtOH can potentiate the longer-term effects of high concentrations of Ca2+ on DNA synthesis whereas, in keratinocytes, EtOH may inhibit Ca2+-induced differentiation. PMID- 10371397 TI - Chronic ethanol exposure enhances signaling through muscarinic receptors expressed by cRNA injection in Xenopus oocytes: implications for mechanism of action. AB - The molecular mechanisms underlying the cerebral symptoms of ethanol withdrawal syndrome are poorly understood. In addition to ethanol's effect on GABA and NMDA receptors, ethanol affects muscarinic acetylcholine signaling. This interaction has attracted attention because of the importance of muscarinic signaling in consciousness. Chronic ethanol exposure increases muscarinic receptor binding. Increased transcription of receptor message has been suggested as the underlying mechanism, but this hypothesis has not been tested directly. Therefore, we studied the effects of ethanol on muscarinic signaling in a model that bypasses transcription of muscarinic receptor genes. We expressed rat m1 muscarinic receptors by cRNA microinjection in Xenopus oocytes. Cells were voltage-clamped at -70 mV and effects of prolonged (24, 48, and 72 hr) exposure to ethanol (25, 50, and 100 mM) on methylcholine-induced calcium-activated Cl- currents were determined. Effects of prolonged ethanol exposure on currents induced by stimulation of lysophosphatidate receptors, direct G protein activation, or inositol trisphosphate receptor activation were studied as well. Prolonged ethanol exposure enhanced methylcholine (or lysophosphatidate-)-induced currents in a time- and concentration-dependent manner. Thus, enhanced muscarinic gene transcription is not required for ethanol enhancement of muscarinic signaling. Lack of ethanol effect on inositol trisphosphate-induced signaling suggests that intracellular signaling systems downstream of phospholipase C are not involved. In contrast, currents induced by direct G protein stimulation were enhanced significantly. Therefore, one potential site of ethanol's action on muscarinic signaling is upregulation of the associated G protein or enhancement of its functioning. PMID- 10371398 TI - Acute lead acetate administration potentiates ethanol-induced locomotor activity in mice: the role of brain catalase. AB - It has been proposed that brain catalase plays a role in the modulation of some psychopharmacological effects of ethanol. The acute administration of lead acetate has demonstrated a transient increase in several antioxidant cell mechanisms, including catalase. In the present study, we investigated the effects of acute lead acetate administration on ethanol-induced behavior, brain catalase activity, and the relation between both effects. Lead acetate (100 mg/kg) or saline was injected intraperitoncally in mice. At different intervals of time (1, 3, 5, 7, 9, or 11 days) after this treatment, ethanol (2.5 g/kg) was injected intraperitoneally and the mice were placed in open field chambers. Results indicated that the locomotor activity induced by ethanol was significantly increased. Maximum ethanol-induced locomotion increase (70% more activity than control animals) was found in animals treated with lead acetate 7 days before ethanol administration. Total brain catalase activity in lead-pretreated animals also showed a significant induction, which was maximum 7 days after lead administration. A significant correlation was found between both effects of locomotor and catalase activity. In a second study, the effect of lead administration on d-amphetamine (2.0 mg/kg) and tert-butanol-(0.5 g/kg) induced locomotor activity was investigated. Lead acetate treatment did not affect the locomotion induced by these drugs. These data suggest that brain catalase is involved in ethanol's effects. They also provide further support for the notion that acetaldehyde may be produced directly in the brain via catalase and that it may be a factor mediating some of ethanol's central effects. PMID- 10371399 TI - Effects of L-type voltage-sensitive calcium channel modulators on the discriminative stimulus effects of ethanol in rats. AB - The purpose of the present investigation was to determine whether administration of dihydropyridine-sensitive calcium channels plays a role in modulating the discriminative stimulus effects of ethanol. A food-reinforced operant methodology was used to train adult male Long-Evans rats to discriminate either 1.0 g/kg of ethanol from water or 2.0 g/kg of ethanol from water. After training, two sets of experiments were conducted. First, a time course procedure was implemented whereby a single intraperitoneal dose of either nimodipine (3, 10, 30 mg/kg), nifedipine (3, 10, 30 mg/kg), or isradipine (1, 3, 10, 17 mg/kg) was administered, and test sessions were conducted 10, 20, 30, 60, and 90 min postinjection. Complete substitution (80% or greater ethanol-appropriate responding) for ethanol by these dihydropyridine compounds varied among subjects with dose and pretreatment time. Overall, isradipine substituted for the discriminative stimulus effects of ethanol in the greatest percentage of animals in both training groups. However, substitution varied with dose. Nifedipine dose dependently substituted for ethanol in half of the animals trained with 1.0 g/kg of ethanol but was less effective in animals trained with 2.0 g/kg of ethanol. For the second set of experiments, a single dose of nimodipine, nifedipine, isradipine, or (-)-BAY k 8644 was administered before determination of the cumulative ethanol dose response. Nifedipine produced a significant leftward shift and (-)-BAY k 8644 produced a significant rightward shift in the ethanol dose-response curve in animals trained to discriminate 2.0 g/kg of ethanol from water. These results indicate that the administration of VGCC modulators plays an indirect role in the discriminative stimulus effects of ethanol. PMID- 10371400 TI - Alcohol effects on mood, equilibrium, and simulated driving. AB - BACKGROUND: The effects of alcohol on simple versus complex psychomotor performance were compared in 18 adults. METHODS: Subjects received ethanol doses of 0.0, 0.5, and 0.8 g/kg in a randomized, double-blind, within-subject design. Forty minutes after finishing their drinking, the subjects completed a 60-min battery of tests that included: 1) a sensory organization posturography test (EquiTest); 2) latency to apply the brake after appearance of a barrier in a driving simulator (brake reaction time); 3) visual analog subjective-effects scales (VAS); 4) the Profile of Mood States (POMS); 5) critical flicker fusion (CFF); and 6) choice reaction time (CRT). RESULTS: Alcohol dose dependently reduced composite equilibrium scores and increased brake reaction time. On the CRT task, total reaction time was significantly increased after the high dose but not the low dose. Alcohol dose dependently increased VAS "dizzy," "high," and "drug effect" ratings. The POMS and CFF were not significantly affected by alcohol. CONCLUSIONS: These data suggest that an ethanol dose that neither influences certain mood states nor impairs simple psychomotor task performance nonetheless may impair equilibrium and complex psychomotor tasks (e.g., driving). PMID- 10371401 TI - Acute and long term effects of buspirone treatments on voluntary ethanol intake in a rat model of alcoholism. AB - Buspirone, a 5-HT1A agonist, has been shown to decrease the intake of ethanol when given as a single dose to rats with a psychological dependence induced according to our rat model of alcoholism. The present experiment evaluates the effects different treatments with buspirone have on voluntary ethanol intake in these psychologically dependent rats. As a first treatment, buspirone was given once daily for 23 days at the dose of 20 mg/kg/day. Ethanol was withheld except for the first and the last day of the treatment. On the first day, the buspirone injection decreased ethanol intake from the pretreatment value (1.94+/-0.18 g/kg/day), down to 1.36+/-0.18 g/kg (p < 0.01, n = 12). The rats were again given a choice between water and 10% ethanol after the last injection of buspirone. During the following 24 hr period, the ethanol intake was increased to 3.56+/ 0.24 g/kg/day (p < 0.001 vs. the pretreatment intake, n = 12). A loss of correlation with the pretreatment intake of ethanol indicated an altered regulation of ethanol intake for approximately 3 more weeks. Fifteen weeks after the start of the first treatment, buspirone (20 mg/kg) was re-tested as a single dose, with no effect on ethanol intake. Twenty-two weeks after the start of the first treatment, a 1-week treatment with 20 mg/kg/day of buspirone was started. During this treatment, the rats had a continuous choice between 10% ethanol and water. There was, as in the first re-test, no effect on ethanol intake on the first day of the treatment. However, on the last 2 days of the treatment, the ethanol intake was increased to 2.86+/-0.28 g/kg and to 2.89+/-0.26 g/kg respectively (p < 0.05, n = 10 on both days, compared with the pretreatment intake of 1.78+/-0.36 g/kg). Thus, an acute dose of buspirone can decrease voluntary ethanol intake in psychologically dependent rats, but long-lasting changes in the effect of buspirone seem to develop during a 3-week treatment period. PMID- 10371402 TI - Differences in neurophysiological indices of associative learning in alcohol preferring and nonpreferring rats. AB - Alcohol-preferring (P) and -nonpreferring (NP) rats differ in baseline neurophysiological measures as well as in their neurophysiological responses to ethanol. In the present study, these lines of rats were assessed to determine whether they also differ in their neurophysiological responses during an associative learning paradigm. Male P and NP rats were implanted with electrodes in the frontal cortex, parietal cortex, and amygdala. Both groups were then exposed to an associative learning paradigm. During the first five sessions (conditioning phase), an infrequently presented tone was paired with the delivery of a food pellet. A second tone was also presented during these sessions, but this tone was never paired with food pellet presentation. During the second five sessions (extinction phase), neither of the tones were paired with food pellet presentation. Event-related potentials (ERPs) in response to the tones were recorded during both phases of the experiment. During the first session, the latency of the N1 and P3 waves from the cortical lead in response to the food paired tone was significantly longer in the NP rats than in P rats. In addition, P rats displayed significant increases in the latency of the P2 wave component in the cortex and the P3A wave component in the amygdala in response to changes in the association between food pellet and tone presentation. These data indicate that the P rats were more responsive to changes in the association between food pellet delivery and tone presentation. They also suggest more enhanced associative learning in P rats than in NP rats. This enhanced learning could be an innate trait of P rats or the result of altered learning due to differences in anxiety between P and NP rats. PMID- 10371403 TI - Prospective study of alcohol consumption patterns in relation to symptomatic gallstone disease in men. AB - Although the association between alcohol intake and gallstone disease has been characterized previously, the relation between alcohol consumption patterns, specific types of alcoholic beverages, and risk for cholelithiasis has not been addressed systematically. These issues were examined prospectively in a cohort of men who were free from symptomatic gallstone disease in 1986 and were followed to 1996. During follow-up, 2.4% of the men reported newly symptomatic gallstones that were diagnosed by ultrasonography or x-ray, or a cholecystectomy. After adjusting for other known or suspected risk factors, an increase in the amount of alcohol consumed was associated with a decreased risk of symptomatic gallstone disease. An increase in frequency of alcohol consumption also was related to decreased risk. Combining the reports of quantity and frequency of alcohol intake, a consumption pattern that reflected frequent intake (5-7 days/week) of any given amount of alcohol was associated with a decreased risk, as compared with nondrinkers. In contrast, infrequent alcohol intake (1-2 days/week) showed no significant association with risk. All alcoholic beverage types were inversely associated with risk of symptomatic gallstone disease, independent of patterns of consumption. These results suggest that frequent, moderate intake of alcohol decreases the risk for symptomatic gallstone disease, in contrast to infrequent or episodic alcohol intake. Recommendations regarding the benefit of moderate quantities of alcohol for gallstone discase should be weighed against the potential health hazards of alcohol consumption. PMID- 10371404 TI - Ethanol feeding does not affect the efficacy or pharmacokinetics of azithromycin, trovafloxacin, or ceftriaxone in a rat model of pneumococcal pneumonia. AB - A rat model of ethanol feeding was used to study the effects of ethanol on antibiotic therapy of pneumococcal pneumonia. Male Sprague-Dawley rats (150 g) received a liquid diet containing 36% of total calories as ethanol. Controls were pair-fed a liquid diet without ethanol or received rat chow. Diets began 7 days pre- and continued postinfection. Rats were infected transtracheally with type 3 Streptococcus pneumoniae and then treated with azithromycin (50 mg/kg), trovafloxacin (50 mg/kg), or ceftriaxone (100 mg/kg) injected subcutaneously twice daily for 5 days. Antibiotic levels in serum, lung cells, and lavage fluid were measured by HPLC. Ethanol- and pair-fed rats had depressed baseline peripheral neutrophil counts but were able to generate adequate numbers of peripheral and pulmonary polymorphonuclear leukocytes early in the course of their infection. Ethanol feeding did not alter the pharmacokinetics of azithromycin, trovafloxacin, or ceftriaxone. All three antibiotics were equally effective in curing experimental pneumococcal pneumonia, and survival rates were similar in treated ethanol-fed and control rats. PMID- 10371405 TI - Effect of voltage-dependent calcium channel blockers on ethanol-induced beta endorphin release from hypothalamic neurons in primary cultures. AB - The voltage-dependent calcium channel (VDCC) has been shown to mediate calcium entry into neurons that regulates neurotransmission in many neuronal cells. Four major types of VDCCs (three high-voltage-activated L-, N-, and P-types and one low-voltage-activated T-type) have been identified in neurons. Involvement of the VDCC in ethanol-stimulated beta-endorphin (beta-EP) release from hypothalamic neurons has not been studied. In the present study, the role of VDCC on basal and ethanol-induced beta-EP release was determined by using rat fetal hypothalamic cells in primary cultures. Treatments with a 50 mM dose of ethanol for 3 hr increased immunoreactive beta-EP (IR-beta-EP) release from hypothalamic cells maintained in cultures for 9 days. Ethanol-induced IR-beta-EP release was inhibited by a P/Q-type channel blocker omega-agatoxin TK (0.1-1 microM), an N type channel blocker omega-conotoxin (0.1-1 microM), an L-type blocker nifedipine (1-10 microM), and a T-type blocker flunarizine (1-10 microM). The minimal effective doses of these blockers that blocked the ethanol response produced no significant effects on basal release of IR-beta-EP; neither did these doses of the blockers produce any significant effects on cell viability. These results suggest that ethanol-stimulated IR-beta-EP release is regulated by extracellular calcium involving P-, N-, L- and T-type channels. PMID- 10371406 TI - Immune dysfunction during alcohol consumption and murine AIDS: the protective role of dehydroepiandrosterone sulfate. AB - Acquired immune deficiency syndrome (AIDS) is a clinical disorder caused by the human immunodeficiency virus (HIV) after development of severe immunosuppressive changes. Chronic ethanol (EtOH) consumption accentuates the severity of murine AIDS (MAIDS). Because hormone production is often suppressed by chronic EtOH intake, as well as retrovirus infection, we investigated whether hormone supplementation during chronic EtOH consumption contributes to slowing immune dysfunction caused by LP-BM5 infection and/or EtOH use. Because dehydroepiandrosterone sulfate (DHEAS) was previously shown to have immune enhancing properties during MAIDS, we determined whether DHEAS reduced cytokine dysregulation otherwise exacerbated by chronic EtOH intake during MAIDS. Adult female C57BL/6 mice were infected with LP-BM5 murine retrovirus. Some were fed 40% EtOH in drinking water and agar gel for 16 weeks postinfection. EtOH consumption further inhibited T- and B-cell proliferation beyond suppression due to retrovirus infection. Interleukin (IL)-2 release produced by concanavalin A stimulated splenocytes was reduced by EtOH use by infected and uninfected mice. DHEAS overcame much of the effects induced by retrovirus infection and/or EtOH use. IL-4 secretion and IL-6 secretion were enhanced. Hepatic vitamin E levels were decreased by murine retrovirus infection, as well as by EtOH use in both uninfected and infected mice. In addition, DHEAS (0.01%) supplementation during MAIDS prevented the further dysregulation of cytokines and hepatic lipid peroxidation due to EtOH intake, partially restored T- and B-cell proliferation, and maintained hepatic vitamin E levels to near normal levels. PMID- 10371407 TI - Prenatal alcohol use and offspring size at 10 years of age. AB - The Maternal Health Practices and Child Development Project is a longitudinal study of the effects of prenatal exposure to alcohol and other substances. Women were selected from a prenatal clinic and interviewed at the 4th and 7th months of pregnancy. Their offspring were examined at delivery, at 8 and 18 months, and at 3, 6, and 10 years. This report examined 610 offspring, at age 10, who were exposed prenatally to alcohol. Most alcohol use in this low-income cohort was light to moderate, although the entire spectrum of alcohol use is represented. The weight, length, head circumference, and skinfold thickness of the offspring were measured. At each assessment phase, we found a significant association between size and prenatal exposure to alcohol. At age 10, the children who were prenatally exposed to alcohol continued to be significantly smaller in weight, height, head circumference, and skinfold thickness. These results indicate that prenatal alcohol exposure has a long-term impact on offspring growth. PMID- 10371409 TI - Ethanol suppression of the functional state of polymorphonuclear leukocytes obtained from uninfected and simian immunodeficiency virus infected rhesus macaques. AB - BACKGROUND: Human immunodeficiency virus (HIV) infection and alcohol abuse frequently coexist in the host and are known to suppress individually the host response to a variety of opportunistic infections. METHODS: This study examined the effects of in vitro ethanol exposure on several functions of polymorphonuclear leukocytes (PMNs) that were obtained from uninfected and simian immunodeficiency virus (SIV)-infected rhesus macaques, at the asymptomatic and terminal stages of infection. RESULTS: The PMNs obtained from rhesus macaques at both the asymptomatic and terminal stage of SIV disease had elevated phagocytic activity and increased CD11b expression compared with PMNs from uninfected animals. In vitro 100 mM ethanol suppressed phagocytosis and CD11b adhesion molecule expression by PMNs, regardless of the stage of SIV infection. Treatment of PMNs with granulocyte colony-stimulating factor (G-CSF) attenuated the inhibitory effect seen with prior ethanol exposure. CONCLUSIONS: These data demonstrate that the functional state of PMNs from uninfected as well as SIV infected rhesus macaques is impaired by direct exposure to intoxicating concentrations of ethanol and that this effect can be attenuated by G-CSF. If alcohol intoxication similarly suppressed PMN function in vivo, it would further increase susceptibility of these hosts to secondary infections. Furthermore, G CSF may be useful in overcoming the suppressive effects of ethanol on PMN function in such patients. PMID- 10371408 TI - Effects of Nitric oxide synthase blockade on the acute response of the reproductive axis to ethanol in pubertal male rats. AB - The effects of ethanol (EtOH) and nitric oxide (NO) are well known in the adult male rat reproductive axis. In the present study, we investigate the effects of EtOH, NO, and their interaction on key genes and reproductive hormone levels in mid- (45-day) and late pubertal (55-day) male rats. Using three different NO synthase blockers--N'omega-nitro-L-arginine methyl ester (L-NAME), N'omega-nitro L-arginine (L-NA), and 7-nitroindazole--we show that it is possible to block, in part, some of the disruptive effects of EtOH. L-NAME totally prevented the EtOH induced fall in serum testosterone in both 45- and 55-day-old rats (p < 0.05 and p < 0.001, respectively). On the other hand, the D-NAME, an inactive isomer of L NAME, did not protect testosterone from suppression caused by EtOH. Similarly, L NA and 7-nitroindazole prevented the suppression of testosterone caused by EtOH in 55-day-old animals (p < 0.001 L-NA and p < 0.05 for 7-nitroindazole), but not in the 45-day-old rats. Serum luteinizing hormone (LH) was significantly reduced by EtOH in all the studies in both age groups. L-NAME (but not D-NAME) and L-NA prevented this inhibition in 55-day-old animals (p < 0.001 for L-NAME and p < 0.01 for L-NA). However, only L-NA was able to prevent the effects of EtOH on LH in the 45-day-old rats. 7-Nitroindazole was unable to prevent the decrease in LH in either age group. Despite changes in the other reproductive hormones, there were no consistent changes in hypothalamic concentrations of either LH releasing hormone (LHRH) or its precursor, pro-LHRH. No treatment caused any change in steady-state levels of beta-LH mRNA. There were no consistent changes in pro-LHRH mRNA; but, interestingly, in 45-day-old rats, L-NA given with or without EtOH lead to a significant fall in LHRH gene expression. Our findings indicate that the acute suppressive effects of EtOH on the hypothalamic-pituitary-gonadal axis of the pubertal male rat can be at least partially prevented by NO synthase blockade. PMID- 10371410 TI - Effect of acute ethanol administration on toloxatone pharmacokinetics in rabbits. AB - The pharmacokinetics of toloxatone and ethanol were determined in plasma and cerebrospinal fluid of conscious rabbits. According to a cross-over design, rabbits (n = 5) randomly received on three separate occasions either toloxatone (5 mg/kg), ethanol (1 g/kg), or toloxatone and ethanol (5 mg/kg and 1 g/kg, respectively) by intravenous injection. Toloxatone and ethanol concentrations were measured by HPLC with UV detection and GC with flame ionization detection, respectively. Ethanol concentration profiles in plasma were characterized by a rapid decline occurring within the first 30 min after administration followed by a linear (zero-order) phase that persisted for the length of the experiment. The maximum ethanol elimination rate was 0.31+/-0.20 g/h x kg. Toloxatone concentrations in plasma and cerebrospinal fluid were characterized by a multiexponential decay with effective half-lives of 0.39+/-0.06 and 0.56+/-0.07 hr, respectively. Toloxatone passage through the blood brain barrier was rapid and important. Our results also demonstrate that acute ethanol administration had no effect on toloxatone pharmacokinetics and that toloxatone administration had no effect on ethanol pharmacokinetics. PMID- 10371411 TI - Glucocorticoid fast feedback is not altered in rats prenatally exposed to ethanol. AB - Animals exposed in utero to ethanol exhibit hormonal hyperresponsiveness to stressors in adulthood. One possible mechanism for this hyperresponsiveness is a deficit in negative feedback regulation of the hypothalamic-pituitary-adrenal axis. The present study tested the hypothesis that a deficit in the fast feedback time domain may play a role in the hormonal hyperresponsiveness in ethanol exposed rats. Sprague-Dawley offspring from prenatal ethanol (E), pair-fed (PF), and ad lib-fed control (C) groups were tested in two experiments. Experiment 1 used a swim stress paradigm and tested animals at the trough of the corticosterone (CORT) circadian rhythm. Experiment 2 used ether stress and tested animals at the peak of the circadian rhythm. Animals were injected subcutaneously with CORT or saline and were immediately subjected to either a 5-min swim stress or a 1-min ether stress. Half the animals were terminated immediately after stress (5-min postinjection), and the rest were terminated 25 min later. Plasma levels of CORT and ACTH were assayed to determine whether E animals differed from control animals in showing a CORT-induced blunting of the ACTH response to the stressor, indicating alterations in fast feedback regulation. Injection of CORT significantly blunted the ACTH response to swim stress (experiment 1) in E, PF, and C females and males, compared with their saline-injected counterparts. There were no significant differences among groups. Similarly, CORT-injected males in E, PF, and C groups all exhibited a significantly blunted ACTH response to ether stress (experiment 2). CORT-injected C females also exhibited a significantly blunted ACTH response to ether stress, whereas E females showed an obvious CORT decrease that approached significance. However, PF females showed a clear deficit in fast feedback regulation. Together, these data suggest that: (1) CORT injection can serve as a fast feedback signal that can blunt the ACTH response to a stressor; and (2) prenatal ethanol exposure does not produce a deficit in hypothalamic-pituitary-adrenal feedback regulation in the fast feedback time domain. PMID- 10371412 TI - Stellate cell activation in alcoholic fibrosis--an overview. AB - There has been remarkable progress in our understanding of how chronic alcohol ingestion may lead to hepatic injury and scarring, or fibrosis. Hepatic fibrosis represents the liver's wound healing response and is characterized by accumulation of interstitial matrix, or scar. Fibrosis in the liver results from the activation of stellate cells, or resident mesenchymal cells. Stellate cell activation is a dramatic phenotype transition whose net effect is the replacement of normal liver matrix by scar. Features of stellate cell activation include increased cell accumulation from proliferation and directed migration, increased matrix production, enhanced contractility, accelerated degradation of the normal liver matrix, release of profibrogenic cytokines, and loss of cellular vitamin A. Alcohol may enhance fibrogenesis through stimulation of stellate cells by hypoxia, generation of lipid peroxides from damaged hepatocytes, production of acetaldehyde that may have direct fibrogenic activity, and through activation of Kupffer cells or resident macrophages. Unanswered questions remain to be studied, but the clarification of underlying mechanisms of fibrosis portends continued progress in our ability to treat alcoholic liver fibrosis. PMID- 10371413 TI - Cytokine regulation of hepatic stellate cells in liver fibrosis. AB - Cytokines constitute a major class of mediators responsible for "activation" of hepatic stellate cells (HSCs) in vitro and in vivo. They are largely divided into mitogenic (transforming growth factor-alpha, platelet-derived growth factor, interleukin-1, tumor necrosis factor-alpha, and insulin-like growth factor) and fibrogenic (transforming growth factor-beta and interleukin-6) cytokines. In addition to their mitogenic (stimulation of cell proliferation) and fibrogenic (induction of matrix proteins) properties, they are also shown to confer in vitro unique cellular changes known to be the key features of HSC "activation," including loss of vitamin A, stimulation of migration, enhanced cellular contractility, and matrix metalloproteinase and tissue inhibitor of metalloproteinase induction. Potential cellular sources of the cytokines consist of hepatic macrophages, endothelial cells, biliary epithelial cells, lymphocytes, platelets, hepatocytes, and activated HSCs. To better understand the mode of actions and the pathogenetic significance of cytokines/chemokines involved in "activation" of HSCs, the following four questions need to be addressed: (1) What other cytokines are expressed by HSCs to establish critical autocrine stimulation? (2) What are endogenous or exogenous priming factors for HSC stimulation? (3) What is the mechanism of activation for transforming growth factor-beta, the pivotal fibrogenic cytokine? (4) How important are HSC-derived proinflammatory mediators in liver fibrosis? This review will discuss these questions, along with the current understanding of the role of cytokines in HSC activation. PMID- 10371414 TI - Leukocytes as modulators of stellate cell activation. AB - Activation of stellate cells is central to the process of hepatic fibrogenesis. Stellate cell activation can be influenced by many factors, including cytokines, oxidants, and alterations in the perisinusoidal extracellular matrix. These factors can be produced by resident liver cells (hepatocytes, Kupffer cells, or stellate cells themselves); however, infiltrating leukocytes may also play an important role. Because liver fibrosis often follows a prolonged period of hepatic inflammation, investigators have begun to study leukocytes as modulators of stellate cell activation. The following data summarize recent investigations in this area that focus on neutrophils as well as mononuclear cells. PMID- 10371417 TI - Acetaldehyde-mediated collagen regulation in hepatic stellate cells. AB - Alcohol-induced liver cirrhosis is one of the major causes of death worldwide. Strong evidence has established that ethanol's first metabolite, acetaldehyde, is highly fibrogenic and enhances the deposition of many extracellular matrix components by hepatic stellate cells. This article reviews our current knowledge of the molecular mechanisms whereby acetaldehyde induces these activities, with particular emphasis on those related to the upregulation of type I collagen. PMID- 10371415 TI - Intracellular signaling pathways in stellate cell activation. AB - Pathological fibrogenesis in the liver is mediated by activated stellate cells. These cells have a myofibroblastic phenotype with the ability to proliferate and synthesize large quantities of extracellular matrix components. A number of factors have been proposed to initiate and perpetuate the fibrogenic process in stellate cells, including inflammatory cytokines, alterations in the extracellular matrix, growth factors, and oxidative stress. Some recent research has focused on the intracellular signaling pathways that are stimulated by these factors in stellate cells, including mitogen-activated protein kinases, phosphatidylinositol 3-kinase, focal adhesion kinase, and protein kinase C. This paper will summarize the experimental evidence that implicates these pathways in stellate cell activation, focusing on the effects of exposure to platelet-derived growth factor, tumor necrosis factor-alpha, and fibronectin. Implications for alcohol-induced hepatic fibrosis and future directions for research will also be discussed. PMID- 10371416 TI - Role of transcriptional factors in stellate cell activation. AB - Fibrogenic stimuli induce a complex activation process in the hepatic stellate cell (HSC) resulting in numerous changes in gene expression. These changes include increases in the synthesis and deposition of several extracellular matrix (ECM) proteins, most notably type I collagen. The synthesis of type I collagen following HSC activation is controlled, in part, at the transcriptional level. Dramatic changes occur in transcription factor binding to ECM genes following HSC activation. Similarities between type I collagen and biglycan gene expression suggest that common transcriptional mechanisms may regulate ECM expression following HSC activation. Several transcription factors display increased activity with HSC activation. The identification of new potential therapeutic targets may become evident providing novel therapeutic avenues aimed at either blocking or slowing the fibrogenic or inflammatory process in which the activated HSC participates. PMID- 10371418 TI - Role of metalloproteinases in liver fibrosis. AB - Collagen deposition in the cirrhotic liver is the result of an imbalance between the amount of collagen produced and that, which is degraded. Although several groups have actively investigated the mechanisms that regulate collagen gene expression in the liver, little is known regarding those involved in the regulation of interstitial collagenases. In this study, we shall express our personal ideas regarding the role of metalloproteinases in collagen degradation in the cirrhotic liver, with special emphasis on the interstitial collagenases and some factors that may limit collagen degradation in vivo. PMID- 10371419 TI - Tissue inhibitors of metalloproteinases: role in liver fibrosis and alcoholic liver disease. AB - In liver fibrosis, activated hepatic stellate cells (HSC) play a major role in the deposition of excess extracellular matrix, including fibrillar collagens type I and type III. In addition to matrix protein synthesis, HSC regulate matrix degradation in the liver. This is mediated via a combination of synthesis of matrix (pro)metalloproteinases, which activate these zymogens via specific mechanisms and by inhibiting the active matrix-degrading enzymes via expression of tissue inhibitors of metalloproteinases (TIMPs). There are currently four members of the TIMP family described and of these, both TIMP-1 and TIMP-2 are synthesised by HSC. These observations have led to the suggestion that inhibition of matrix degradation mediated by a change in HSC-expression of TIMPs relative to metalloproteinases, such as interstitial collagenase, may contribute to progression of liver fibrosis. This hypothesis is supported by studies of human liver disease in which TIMP-1 expression is upregulated 5-fold in cirrhotic compared with normal liver. TIMP-1 and TIMP-2 expression is also upregulated in animal models of progressive fibrosis, whereas expression of collagenase is unchanged. In a model which is characterized by natural resolution of liver fibrosis, degradation of the deposited fibrillar liver matrix is accompanied by rapid down-regulation of TIMP-1 expression. In alcoholic liver disease, the role of TIMPs has not been studied exhaustively, but the evidence currently available supports a role for inhibition of matrix degradation by TIMPs in this progressive fibrotic liver disease. PMID- 10371420 TI - Prevention and treatment of liver fibrosis based on pathogenesis. AB - Multiple agents have been proposed for the prevention and treatment of fibrosis. S-adenosylmethionine was reported to oppose CCl4-induced fibrosis in the rat, to attenuate the consequences of the ethanol-induced oxidative stress, and to decrease mortality in cirrhotics. Anti-inflammatory medications and agents that interfere with collagen synthesis, such as inhibitors of prolyl-4-hydroxylase and antioxidants, are also being tested. In nonhuman primates, polyenylphosphatidylcholine (PPC), extracted from soybeans, protected against alcohol-induced fibrosis and cirrhosis and prevented the associated hepatic phosphatidylcholine (PC) depletion by increasing 18:2 containing PC species; it also attenuated the transformation of stellate cells into collagen-producing transitional cells. Furthermore, it increased collagen breakdown, as shown in cultured stellate cells enriched with PPC or pure dilinoleoyl PC, the main PC species present in the extract. Because PPC and dilinoleoyl PC promote the breakdown of collagen, there is reasonable hope that this treatment may be useful for the management of fibrosis of alcoholic, as well as nonalcoholic, etiologies and that it may affect not only the progression of the disease, but may also reverse pre-existing fibrosis, as demonstrated for CCl4-induced cirrhosis in the rat and as presently tested in an ongoing clinical trial. PMID- 10371421 TI - Targeting of therapeutics to the liver: liposomes and viral vectors. AB - A successful strategy for the treatment of hepatic damage should be based on the use of nontoxic drugs with a high therapeutic index and safety profile, as well as selective targeting delivery. To achieve these goals, several biological and synthetic vehicles have been formulated with a wide range of chemical and physical properties. These vectors allow the delivery of either traditional drugs or genetic material to the cells. Despite remarkable progress, the design of an ideal carrier is still to be accomplished. Safety issues concerning viral vectors and low efficiency of synthetic delivery systems are the main limitations of present vectors. Once these problems are overcome, the liver-specific targeting systems will provide new opportunities to treat a vast range of hepatic diseases. PMID- 10371422 TI - TMJ disorders complicate treatment planning. PMID- 10371423 TI - Long-term stability of mandibular orthopedic repositioning. AB - Mandibular anterior repositioning appliances attempt to diminish temporomandibular joint pain, soft tissue noise, and myofascial discomfort by altering condyle-disc relationships. Secondary stabilization of the occlusion to this arbitrary anterior position through orthodontic tooth movement may significantly alter functional and muscular relationships. A case report is illustrated to show that as the functional environment attempted to reestablish equilibrium through adaptation, relapse occurred as the condyles "seated" posteriorly and superiorly toward their original relationship within the fossa. For all practical purposes, complete relapse of the orthodontic treatment result took place over time. PMID- 10371424 TI - Case report: treatment for a patient with a history of TMJ disorder. AB - Establishing a knowledge-based protocol for the treatment of orthodontic patients who report a history of temporomandibular dysfunction can alert the practitioner to potential treatment pitfalls before they happen. While the joints can be extremely adaptive, some individuals are subject to painful and/or limited function. Others have acquired condylar positions that, if not recognized, could lead to serious alterations in the original treatment plan. Combining a thorough diagnostic protocol with a therapeutic regimen that seeks to establish a stable condylar and occlusal position-prior to initiating treatment- is essential. PMID- 10371425 TI - Putting nickel titanium wire in its place. PMID- 10371426 TI - Finite-element morphometry of soft tissue morphology in subjects with untreated Class III malocclusions. AB - Soft tissue dynamics may contribute to maxillomandibular allometry (size-related changes in shape) associated with the development of Class III malocclusions. Lateral cephalographs of 124 prepubertal European American children were traced and 12 soft tissue landmarks were digitized. Resultant geometries were normalized, and Procrustes analysis established the statistical difference (p<0.001) between mean Class III and Class I configurations. Comparing the Class III configurations with normals for size-change, color-coded finite element analysis revealed a superoinferior gradient of positive allometry of the Class III facial nodal mesh. A conspicuous area of negative allometry (approximately 40%) was localized near soft subspinale, with a approximately 70% increase in size in the mental region. For shape-change, the Class III facial mesh was isotropic, except in the anisotropic circumoral regions. Conventional cephalometry revealed that about 50% of linear and 75% of angular parameters differed statistically (p<0.001). Soft tissue dynamics during early postnatal development may contribute to the development of Class III malocclusions. PMID- 10371427 TI - Evaluation of differential growth and orthodontic treatment outcome by regional cephalometric superpositions. AB - Cephalometric superimposition on cranial base is the accepted method for evaluating mandibular displacement during orthodontic treatment and/or growth. However, assessing mandibular position relative to the maxillary base may yield different information. The aim of this study was to evaluate the effects of regional superpositions (cranial versus maxillary) on interpreting mandibular displacement. Both methods were applied to pre- and posttreatment cephalograms of 22 growing children (12 female, 10 male) treated for Class II Division 1 malocclusion. Differences in linear and angular measurements of three mandibular landmarks (pogonion, gnathion, menton) between cranial and maxillary superpositions were statistically significant (p = 0.0001). Vertical displacement of these landmarks contributed significantly to the differences (p = 0.0001). The contribution of horizontal displacement was not statistically significant. The results support the proposition that, in growing children, posttreatment displacement of mandibular skeletal and dental components should be assessed by both maxillary and cranial base superimpositions. The maxilla is subject to rotational and translational changes during growth that may affect the position of the mandible relative to the maxilla in a way inconsistent with the mandibular displacement perceived upon cranial superposition. Since occlusion is directly associated with the positions of the maxillary and mandibular basal bones, the positions of these bones relative to each other is critical in assessing occlusal changes in individual patients. PMID- 10371428 TI - Comparison of preferences in lip position using computer animated imaging. AB - The objectives of this study were to examine the esthetic preferences of lip position in males and females, and to compare them with each other and with a common orthodontic standard using a custom computer animation program. The sample consisted of 53 young adult subjects, 25 males and 28 females. The sample was divided into orthodontically treated and untreated subjects. ANOVA and Scheffe tests were carried out to determine differences between the responses of the various groups. Also, t-tests were used to compare subjects' responses to a commonly used orthodontic standard (Ricketts' E-line). The results indicated a sex-effect, with females preferring fuller lips than males. Significant differences were also found between orthodontically treated subjects and untreated subjects, with untreated subjects preferring fuller lips. Differences were significant at p<0.05. Furthermore, both males and females preferred lip fullness greater than the Ricketts' values. PMID- 10371429 TI - Dentoskeletal effects and facial profile changes in young adults treated with the Herbst appliance. AB - This prospective Herbst study analyzed the sagittal dental and skeletal changes contributing to Class II correction in young adults. Additionally, the alteration in skeletal and soft tissue convexity occurring during treatment was assessed. Early adolescent subjects in the permanent dentition who had been treated with the Herbst appliance were used for comparison. Lateral headfilms from before and after an average treatment period of 8.5 months for the young adults and 7.1 months for the adolescents were evaluated. All adult and adolescent subjects were treated to either Class I or overcorrected Class I occlusal relationships. In both groups the improvement in sagittal incisor and molar relationships was achieved more by dental changes than by skeletal ones. The amount of skeletal change contributing to overjet and molar correction was smaller in the young adult group (22% and 25%, respectively) than in the early adolescent group (39% and 41%, respectively). Skeletal and soft tissue facial profile convexity was reduced in adults and adolescents. Facial profile improvement did not differ between the two groups. The results of this study revealed that the Herbst appliance is most effective in the treatment of Class II malocclusion in young adults. It is suggested that this treatment method could be an alternative to orthognathic surgery in borderline Class II cases. PMID- 10371430 TI - The dawn of rapid maxillary expansion. AB - The first report of lateral maxillary expansion by separation of the maxilla, written by Angell and published in 1860, was discredited. Applying our present day knowledge of the technique to the original documents indicates that the case history agrees in general with current observations. The arguments mounted against Angell, especially by McQuillen, may be dismissed as irrelevant and Angell's thesis is upheld. In addition, good reason exists to accept three further "firsts" in this unprecedented work: (1) The significance of the first permanent molars in occlusal development, (2) the use of a double-action jackscrew, and (3) the use of a retention plate. PMID- 10371431 TI - Effects of a bonded rapid maxillary expansion appliance during orthodontic treatment. AB - The aim of this prospective study was to evaluate changes in the transverse plane following use of an acrylic bonded rapid maxillary expansion (RME) appliance in growing individuals during the active phase of treatment. The sample comprised 14 consecutively treated orthodontic patients (11 girls, 3 boys) who required the use of an RME device on the basis of their individual treatment plans. The mean patient age at the start of treatment was 12.8 years, and the mean overall treatment time was 3.08 years. Seven posteroanterior cephalometric and two dental cast measurements were assessed. Repeated measure analysis of variance and Duncan's multiple range test were used to assess treatment changes. Lower nasal and maxillary base widths and angles, and upper intermolar width increased significantly during RME treatment. Upper intermolar and intercanine widths measured from the dental casts also increased significantly. Except for upper intercanine width, all measurements remained constant at the end of orthodontic treatment. The results of this study suggest that dentoskeletal changes in the transverse dimension following the use of an acrylic bonded RME are maintained satisfactorily at the end of fixed appliance therapy. PMID- 10371432 TI - Relapse after orthodontic correction of maxillary median diastema: a follow-up evaluation of consecutive cases. AB - An evaluation of 96 treated orthodontic patients with maxillary median diastema ranging from 0.50 mm to 5.62 mm (mean 1.22, SD 0.85) was performed 4.0 to 9.0 years after completion of active treatment. Pre- and posttreatment data were gathered from available records. Follow-up data were gathered from records and interviews of 37 patients, and from phone interviews of 59 patients. The incidence of diastema relapse was 49% when scored as either presence of a measurable space at follow-up, a history of orthodontic or prosthetic retreatment to close a reopened space, or continued use of a retainer to control relapse tendency. Logistic regression analysis revealed that pretreatment diastema size and presence of a family member with a similar condition were the only significant risk factors for relapse (p<0.05), while pretreatment spacing in the maxillary anterior dentition approached significance (p = 0.10). No association was found between relapse and presence of an abnormal frenum or an osseous intermaxillary cleft, although patients with an abnormal frenum had a wider pretreatment diastema than those with a normal frenum (p<0.05). Fremitus of the maxillary incisors was the only parameter at follow-up associated with space reopening (p<0.01). PMID- 10371433 TI - Longitudinal occlusal changes from primary to permanent dentition in children with normal primary occlusion. AB - This purpose of this research was to examine the stability of normal occlusion during the transition from primary to permanent dentition. The sample consisted of 128 children (83 boys and 45 girls) 4.5 to 5.5 years old with normal occlusion in the primary dentition. The subjects were reexamined at 12.5 to 13.5 years. None had received orthodontic treatment. Although all the subjects had normal occlusion in the primary dentition, 72.7% (73.5% boys and 71.1% girls) had developed anomalies following eruption of the permanent teeth. These anomalies included crowding, Class II Division 1 or Class II Division 2 malocclusion, mesial occlusion complex, lateral crossbite, anterior crossbite, premature tooth loss, openbite or other anomalies. PMID- 10371434 TI - Combinations of etchants, composite resins, and bracket systems: an important choice in orthodontic bonding procedures. AB - The objectives of this investigation were: (1) to compare the shear bond strengths (SBS) of metal, ceramic, and plastic brackets using different concentrations of maleic and phosphoric acid gels and aqueous solutions, and (2) to determine if a relationship exists between the type of acid etchant and the location of resin after debonding. A sample of 210 bovine incisors was divided among three different bracket groups (Victory series metal, Transcend 6000 ceramic, Spirit MB plastic). Prior to bonding, enamel was acid-etched using 37% phosphoric acid (H3PO4) gel and aqueous solution, 10% maleic acid gel and aqueous solution, 10% H3PO4 gel and aqueous solution, or 2% H3PO4 aqueous solution. SBS testing and the adhesive remnant index (ARI) score provided insight into the effects of the bonding process on enamel. Resin tags associated with each etchant type were inspected under scanning electron microscopy (SEM). Statistical analyses (level of significance, p = 0.05) of the data showed significant differences among groups. It was concluded that specific acid-composite-bracket combinations are recommended for use in clinical orthodontic practice in order to achieve efficient bonding. PMID- 10371435 TI - Optimization of a procedure for rebonding dislodged orthodontic brackets. AB - The purpose of this study was to compare shear bond strength (SBS) of bonded and rebonded orthodontic brackets following a variety of commonly used conditioning treatments and using both light-cured and self-cured composite resin systems. Brackets debonded during the initial determination of SBS were rebonded after the removal of residual resin from enamel surfaces using five different treatments: (1) Remove residual resin using a tungsten carbide bur, re-etch enamel surface, then bond a new bracket; (2) Remove resin from the base mesh with micro-etching then rebond the same bracket, (3) Remove residual resin from the enamel surface using resin-removing pliers, recondition the enamel with an air-powder polisher, then bond a new bracket; (4) Remove residual resin using a rubber cup and pumice, then bond a new bracket; (5) Remove residual resin using pliers alone, then bond a new bracket. The results revealed that the light-cured system produced higher shear bond strength in the initial bond than the self-cured system (p<0.005). Reconditioning the enamel surfaces using a tungsten carbide bur and acid-etching gave the highest SBS (difference 5.8 MPa; p<0.01) and clinically favorable fracture characteristics. The data suggest that the optimal procedure for rebonding dislodged orthodontic brackets is to resurface the enamel using a tungsten carbide bur, acid-etch the enamel, and use a new or re-use an old bracket after microetching. PMID- 10371436 TI - Effect of resin cure mode and fluoride content on bracket debonding. AB - Enamel decalcification around brackets is sometimes observed during and after orthodontic treatment. Reports in the literature suggest that the preventive advantage of fluoride-releasing adhesive resins may be compromised by an increased incidence of bond failure. The purpose of this study was to determine the effects on shear debonding of incorporating fluoride into the bracket bonding system. Another purpose was to determine the effect of polymerization mode on debonding. Orthodontic brackets were bonded to bovine enamel using one of three types of adhesive resin--no-mix, chemically cured, or light-cured--each formulated with and without fluoride. The teeth were stored in artificial saliva for 24 hours or 30 days and then debonded in shear. Data analysis was performed using ANOVA followed by post-hoc multiple comparison between group pairs. It was found that: (1) fluoride had either no effect or it increased the bond value; (2) the no-mix adhesive demonstrated the lowest bond value; (3) the duration of storage in artificial saliva had no effect on the bond value of the chemically cured and light-cured adhesives but did affect the no-mix adhesive; and (4) the no-mix adhesive released significantly less fluoride than the two other products. Thus, the presence of fluoride in the bonding adhesive does not reduce the force required to debond in shear, and chemically or light-cured adhesives provide consistently higher bond values over extended immersion times than the no-mix product. PMID- 10371437 TI - Localized in vivo 1H NMR detection of neurotransmitter labeling in rat brain during infusion of [1-13C] D-glucose. AB - Resolved localized nuclear magnetic resonance (NMR) signals of 1H bound to 13C label in the carbon positions of glutamate C4, C3 and glutamine C4, C3, as well as in aspartate C3, lactate C3, alanine C3, gamma-aminobutyric acid C3, and glucose C1 were simultaneously observed in spectra obtained from rat brain in vivo. Time-resolved label incorporation was measured with a new adiabatic carbon editing and decoupling (ACED) single-voxel stimulated echo acquisition mode (STEAM) sequence. Adiabatic carbon broadband decoupling of 12 kHz bandwidth was achieved in vivo, which decoupled the entire 13C spectrum at 9.4 T. Resonances from N-acetyl-aspartate and creatine were also detected, consistent with natural abundance 13C levels. These results emphasize the potential of 1H NMR for following complex biochemical pathways in localized areas of resting rat brain as well as during focal activation using infusions of 13C-labeled glucose. PMID- 10371438 TI - Low-temperature polarized helium-3 for MRI applications. AB - The first 3He nuclear magnetic resonance (NMR) experiments using low-temperature prepolarization are presented. 3He cells were polarized at 4.2 K and 4.7 T, transported to another magnet, heated to room temperature, and used for NMR experiments at 2.35 T. Cells with and without a rubidium coating were tested. In both cases, the NMR signal was greater than 100 times the thermal equilibrium signal. No evidence of a rubidium coating effect on the longitudinal relaxation time T1 of 3He (500 mbar) at 4.2 K could be demonstrated. NMR gradient-echo images of the cells were acquired. PMID- 10371439 TI - Detection of the early negative response in fMRI at 1.5 Tesla. AB - Recent experimental studies have revealed an initial decrease in magnetic resonance (MR) signal that is consistent with optical imaging results. This initial response, thought to arise from a transient increase in deoxyhemoglobin concentration, is probably more localized to the site of neuronal activation. However, with MR imaging, this early response has only been demonstrated at high fields. In this paper, the observation of the initial response at 1.5 T is reported. Compared with results obtained at 4 T, the present study reveals that the initial response grows more rapidly with the field strength than the late positive response, suggesting a more microvascular origin of the initial response. PMID- 10371440 TI - Multislice perfusion and perfusion territory imaging in humans with separate label and image coils. AB - An arterial spin labeling technique using separate RF labeling and imaging coils was used to obtain multislice perfusion images of the human brain at 3 T. Continuous RF irradiation at a peak power of 0.3 W was applied to the carotid arteries to adiabatically invert spins. Labeling was achieved without producing magnetization transfer effects since the B1 field of the labeling coil did not extend into the imaging region or couple significant power into the imaging coil. Eliminating magnetization transfer allowed the acquisition of multislice perfusion images of arbitrary orientation. Combining surface coil labeling with a reduced RF duty cycle permitted significantly lower SAR than single coil approaches. The technique was also found to allow selective labeling of blood in either carotid, providing an assessment of the artery's perfusion territory. In normal subjects, these territories were well-defined and localized to the ipsilateral hemisphere. PMID- 10371441 TI - Perfusion imaging using FAIR with a short predelay. AB - It is shown theoretically that the flow-sensitive alternating inversion recovery (FAIR) signal intensity difference for flow quantification is independent of the length of the predelay between repeated measurements when assuming complete labeling in between scans. Theory also predicts that flows quantified using the concomitant T1 difference increase significantly with decreasing predelay, because the biexponential relaxation behavior after nonselective inversion is fitted as a monoexponential. The new equations include the effect of the unequal relaxation times of water in tissue (T1) and arterial blood (T1a). While this effect is significant for the signal-difference approach, it is negligible for the T1-difference procedure when using maximum inversion-recovery times shorter than 4 sec. Experiments on cat brain using the FAIR excluding radiation damping (FAIRER) pulse sequence at three different predelays (0.8, 2, and 5 sec) confirm the theoretical predictions. PMID- 10371442 TI - High-resolution magic angle spinning 1H NMR spectroscopic studies on intact rat renal cortex and medulla. AB - High-resolution magic angle spinning 1H NMR (MAS-NMR) spectroscopy was used to investigate the biochemical composition of normal renal cortex and renal papilla samples from rats, and results were compared with those from conventional 1H NMR analysis of protein-free tissue extracts. 1H MAS NMR spectra of samples obtained from inner and outer cortex were found to be broadly similar in terms of metabolite profile, and intra- and inter-animal variability was small. However, the MAS NMR spectra from renal papilla samples were qualitatively and quantitatively different from those obtained from cortex. High levels of free amino acids and several organic acids were detected in the cortex, together with choline, glucose, and trimethylamine-N-oxide. The dominant metabolite resonances observed in papillary tissue were from glycerophosphocholine (GPC), betaine, myo inositol, and sorbitol. On increasing the magic angle spinning rate from 4,200 to 12,000 Hz, the lipid MAS 1H NMR signal profile remained largely unchanged in papillary tissue, whereas "new" resonances from triglycerides appeared in the spectra of cortical tissue, this effect being reversible on returning the spinning rate to 4,200 Hz. Further investigation into the behavior of the lipid components under different spinning rates suggested that the lipids in the cortex were present in more motionally constrained environments than those in the papilla. 1H MAS NMR spectra of tissues are of value both in interrogation of the biochemical composition of whole tissue, and in obtaining information on the mobility and compartmentalization of certain metabolites. PMID- 10371443 TI - Brain metabolic impairment in non-cerebral and cerebral forms of X-linked adrenoleukodystrophy by proton MRS: identification of metabolic patterns by discriminant analysis. AB - Cerebral metabolism in six children with X-linked adrenoleukodystrophy (X-ALD) was studied using 1H magnetic resonance spectroscopy (MRS), and the status of the patients was monitored for evaluating disease progression. Spectra were abnormal even in patients with no cerebral impairment. Four different metabolic patterns were identified, and a metabolic classification of the disease was proposed, from grade 0 to grade III. The evolution of the disease toward grade II appears to be systematic, but many patients did not evolve from this grade to grade III, which is the metabolic mark of severe progressive forms. Metabolic data of X-ALD were processed using discriminant analysis, which provides a classification accuracy of 95.2%. Proton cerebral MRS together with discriminant analysis may be useful during the follow-up in X-ALD for monitoring the evolution of the disease and the effects of therapy. PMID- 10371444 TI - Multiple quantum filtered 23Na NMR spectroscopy of the isolated, perfused rat liver. AB - Isolated, perfused rat livers were examined by single-quantum (SQ) and double quantum-filtered (DQ-filtered) 23Na spectroscopy during prolonged global ischemia and during perfusion with ouabain, low-buffer potassium, or lithium-enriched buffer. Baseline separation of the intracellular (Na(i)+) and extracellular (Na(e)+) sodium resonances using TmDOTP5- allowed a direct comparison of temporal changes in SQ versus DQ-filtered Na(i)+. The SQ Na(i)+ signal increased approximately 150% during the first 15 min of global ischemia and then remained relatively constant over the next 45 min, while the DQ-filtered signal steadily increased approximately 400% over the same 60 min period. In similar experiments in which all perfusate sodium was replaced by lithium, the DQ-filtered Na(i)+ signal increased approximately 180% over a similar period of ischemia. Exposure of livers to ouabain also resulted in larger increases in DQ-filtered versus SQ signal of Na(i)+. The approximately 290% increase in DQ-filtered sodium observed during perfusion of livers with a hypokalemic buffer (1.2 mM K+) could be completely reversed by continued perfusion with a buffer containing normal levels of K+ (4.7 mM). These data suggest that the DQ-filtered Na(i)+ signal of liver does not simply report an increase in [Na(i)+], but may be exquisitely sensitive to other intracellular events initiated by altered physiology. PMID- 10371445 TI - Off-resonance metabolite magnetization transfer measurements on rat brain in situ. AB - Off-resonance metabolite magnetization transfer (MT) experiments were performed on rat brain in vivo and post mortem, with short (18 msec) and long (144 msec) echo-time 1H nuclear magnetic resonance (NMR) spectroscopy. In vivo and post mortem, the methyl protons of total creatine and all protons from glutamate/glutamine showed a strong MT effect on off-resonance saturation, as well as the methyl protons from lactate post mortem. Other resonances, like that of A-acetyl aspartate, showed a much smaller, but detectable, MT effect. The results obtained were confirmed by combining off-resonance saturation with two dimensional correlation spectroscopy. Three water suppression techniques, i.e., presaturation, chemical shift-selective (CHESS), and selective water eliminated Fourier transform (WEFT) were evaluated for their ability to generate an MT effect, to assess their possible influence on metabolite quantification. Presaturation and selective WEFT led to alterations of the total creatine, lactate, and N-acetyl aspartate resonance intensities, while CHESS had no effect. Finally, it was shown that water protons play an important role in the generation of the observed metabolite MT effects. PMID- 10371446 TI - 31P MRS studies of exercising human muscle at high temporal resolution. AB - Methods for measuring mitochondrial activity from 31P magnetic resonance spectroscopy data collected during and after exercise were compared in controls, weight lifters, and peripheral vascular occlusive disease (PVOD) patients. There were trends toward increasing mitochondrial activity during exercise in order from PVOD patients, moderately active controls, highly active controls, to weight lifters. Results from PVOD patients show divergence of some measures due to 1) the non-exponential nature of phosphocreatine recovery, and 2) potential breakdown of [ADP] control of the mitochondria due to lack of oxygen (for Qmax calculation). These results demonstrate the utility of obtaining and directly analyzing high time resolution data rather than assuming monoexponential behavior of metabolite recovery. PMID- 10371448 TI - Sources of variability in the response of coupled spins to the PRESS sequence and their potential impact on metabolite quantification. AB - Using a numerical method of solving the equation of motion of the density matrix, an evaluation is presented of the sources of the marked variability in the response to the point resolved spectroscopy (PRESS) pulse sequence of coupled proton spin systems. The consequences of an inappropriate 180 degrees pulse design and of the limitations on radiofrequency power are demonstrated for a weakly coupled example, lactate. The dominating role of strong coupling, which is present in most brain metabolites, is demonstrated for glutamate, in which 160 terms in the density operator were tracked to monitor the gross changes in lineshape and signal intensity as a function of the two echo times. The predictions of the numerical solutions were confirmed by experiments on phantoms of aqueous metabolite solutions. PMID- 10371447 TI - Determination of relative CMRO2 from CBF and BOLD changes: significant increase of oxygen consumption rate during visual stimulation. AB - The blood oxygenation level-dependent (BOLD) effect in functional magnetic resonance imaging depends on at least partial uncoupling between cerebral blood flow (CBF) and cerebral metabolic rate of oxygen (CMRO2) changes. By measuring CBF and BOLD simultaneously, the relative change in CMRO2 can be estimated during neural activity using a reference condition obtained with known CMRO2 change. In this work, nine subjects were studied at a magnetic field of 1.5 T; each subject underwent inhalation of a 5% carbon dioxide gas mixture as a reference and two visual stimulation studies. Relative CBF and BOLD signal changes were measured simultaneously using the flow-sensitive alternating inversion recovery (FAIR) technique. During hypercapnia established by an end-tidal CO2 increase of 1.46 kPa, CBF in the visual cortex increased by 47.3 +/- 17.3% (mean +/- SD; n = 9), and deltaR2* was -0.478 +/- 0.147 sec(-1), which corresponds to BOLD signal change of 2.4 +/- 0.7% with a gradient echo time of 50 msec. During black/white visual stimulation reversing at 8 Hz, regional CBF increase in the visual cortex was 43.6 +/- 9.4% (n = 18), and deltaR2* was -0.114 +/- 0.086 sec(-1), corresponding to a BOLD signal change of 0.6 +/- 0.4%. Assuming that CMRO2 does not change during hypercapnia and that hemodynamic responses during hypercapnia and neural stimulation are similar, relative CMRO2 change was determined using BOLD biophysical models. The average CMRO2 change in the visual cortex ranged from 15.6 +/- 8.1% (n = 18) with significant cerebral blood volume (CBV) contribution to 29.6 +/- 18.8% without significant CBV contribution. A weak positive correlation between CBF and CMRO2 changes was observed, suggesting the CMRO2 increase is proportional to the CBF increase. PMID- 10371449 TI - Fast magnetic resonance coronary angiography with a three-dimensional stack of spirals trajectory. AB - In this work, three-dimensional (3D) spiral imaging has been utilized for magnetic resonance coronary angiography. Spiral-based 3D techniques can dramatically reduce imaging time requirements compared with 3D Fourier Transform imaging. The method developed here utilized a "stack of spirals" trajectory, to traverse 3D k-space rapidly. Both thick-slab volumes encompassing the entire coronary tree with isotropic resolution and thin-slab volumes targeted to a particular vessel of interest were acquired. Respiratory compensation was achieved using the diminishing variance algorithm. T2-prepared contrast was also applied in some cases to improve contrast between vessel and myocardium, while off-resonance blurring was minimized by applying a linear correction to the acquired data. Images from healthy volunteers were displayed using a curved reformatting technique to view long segments of vessel in a single projection. The results demonstrate that this 3D spiral technique is capable of producing high-quality coronary magnetic resonance angiograms. PMID- 10371451 TI - On the application of susceptibility-weighted ultra-fast low-angle RARE experiments in functional MR imaging. AB - The applicability of displaced, split-echo, and phase-cycled variants of the blood oxygenation level-dependent (BOLD) sensitized ultra-fast low-angle rapid acquisition and relaxation enhancement (UFLARE) technique for the mapping of brain function are examined in functional magnetic resonance imaging (fMRI) experiments at high magnetic field strength (3 T). Activation maps are presented for visual and motor-sensory activation. For the visual studies the range of the stimulation-associated signal intensity changes is 5-7% in voxels containing mainly gray matter and 10-15% in voxels dominated by larger vessels. The motor studies reveal signal changes of 5-10% in the primary motor cortex and in the supplementary motor area. For gray matter, T2* increases from 31.2 +/- 1.5 msec under baseline conditions to 33.0 +/- 1.5 msec during periods of visual stimulation. The results clearly demonstrate that T2*-weighted UFLARE is a robust and reliable method for detection of brain activation. The relative pros and cons of displaced, split-echo, and phase-cycled T2*-sensitized UFLARE versions are discussed for fMRI applications. Since the susceptibility weighting can be freely adjusted from zero upward, the UFLARE variants used are particularly suitable for functional examinations in regions with poor magnetic field homogeneity and at high magnetic field strengths. PMID- 10371450 TI - A numerical approach to non-circular birdcage RF coil optimization: verification with a fourth-order coil. AB - It is demonstrated that birdcage resonators, satisfying conditions of quadrature operation and radiofrequency field homogeneity, can be realized in practice on formers of non-circular cross section described by an equation of the form (x/a)n + (y/b)n = 1 where a and b are constants and n > or = 2 is an integer. Using a ladder network analogous to that of a conventional circular birdcage, optimization algorithms were employed to determine the elemental current distribution on the non-circular cylindrical surfaces. A comparison of circular, elliptical, symmetric and asymmetric fourth-order (n = 4) section birdcage current distributions is presented. A short, asymmetric fourth-order cage was constructed and tested experimentally at 3 T and compared with a conventional circular-section head coil. PMID- 10371452 TI - High spatial resolution EPI using an odd number of interleaves. AB - Ghost artifacts in echoplanar imaging (EPI) arise from phase errors caused by differences in eddy currents and gradient ramping during left-to-right traversal of kx(forward echo) versus right-to-left traversal of kx (reverse echo). Reference scans do not always reduce the artifact and may make image quality worse. To eliminate the need for reference scans, a ghost artifact reduction technique based on image phase correction was developed, in which phase errors are directly estimated from images reconstructed separately using only the forward or only the reverse echos. In practice, this technique is applicable only to single-shot EPI that produces only one ghost (shifted 1/2 the field of view from the parent image), because the technique requires that the ghosts do not completely overlap the parent image. For higher spatial resolution, typically an even number of separate k-space traversals (interleaves) are combined to produce one large data set. In this paper, we show that data obtained from an even number of interleaves cannot be combined to produce only one ghost, and image phase correction cannot be applied. We then show that data obtained from an odd number of interleaves can be combined to produce only one ghost, and image phase correction can be applied to reduce ghost intensity significantly. This "odd number interleaf EPI" provides spatial and temporal resolution tradeoffs that are complementary to, or can replace, those of even-number interleaf EPI. Odd-number interleaf EPI may be particularly useful for MR systems in which reference scans have been unreliable. PMID- 10371453 TI - Correction for EPI distortions using multi-echo gradient-echo imaging. AB - A novel and effective technique is described for distortion correction in echo planar imaging (EPI) utilizing the field maps derived from multi-echo gradient echo images. The distortions from different off-resonance related factors such as field inhomogeneity, eddy current effect, radiofrequency pulse frequency offset, and chemical shift effect can be simultaneously reduced to a great extent. With the proposed post-processing algorithm of multi-channel modulation, distortions may be corrected without unwrapping the phase discontinuities in the derived field map, a process that usually restricts the application of other field map based correction methods. Results from phantom and animal experiments at 4.7 T demonstrate the efficiency of the method in reducing the geometrical distortions in gradient-echo EPI. PMID- 10371454 TI - Quantitative solid-state NMR imaging of synthetic calcium phosphate implants. AB - It is shown that solid-state phosphorus-31 nuclear magnetic resonance imaging can be used to measure quantitatively the mass of hydroxyapatite (HA), a synthetic calcium phosphate used as an orthopedic implant material, in the presence of bone. A three-dimensional projection reconstruction technique was used to produce solid-state images from 998 free induction decays sampled in the presence of a fixed amplitude field gradient whose direction was varied uniformly over the unit sphere. Chemical selection is achieved using T1 contrast, as the synthetic calcium phosphate has a shorter T1 (1.8 sec at 4.7 T) compared with the bone (approximately 15 sec at 4.7 T in vivo, 42 sec ex vivo). Experimental results demonstrating the linear relationship between image intensity and HA density in phantoms containing HA and silicon (IV) oxide, and HA and bone are shown. Chemically pure images of bone mineral and synthetic HA have been computed from images of New Zealand White rabbits acquired in vivo at two different recycle times. The technique can be used to follow noninvasively the resorption and remodeling of calcium phosphate implants in vivo. PMID- 10371455 TI - Resistive homogeneous MRI magnet design by matrix subset selection. AB - A new technique for designing resistive homogeneous multicoil magnets for magnetic resonance imaging (MRI) is presented. A linearly independent subset of coils is chosen from a user-defined feasible set using an efficient numerical algorithm. The coil currents are calculated using a linear least squares algorithm to minimize the deviation of the actual magnetic field from the target field. The solutions are converted to practical coils by rounding the currents to integer ratios, selecting the wire gauge, and optimizing the coil cross-sections. To illustrate the technique, a new design of a short, homogeneous MRI magnet suitable for low-field human torso imaging is presented. Magnets that satisfy other constraints on access and field uniformity can also be designed. Compared with conventional techniques that employ harmonic expansions, this technique is flexible, simple to implement, and numerically efficient. PMID- 10371456 TI - The effect of scanner sound in visual, motor, and auditory functional MRI. AB - The potentially important effect of gradient switching sound on brain function during functional magnetic resonance imaging (fMRI) was studied by comparing experiments with low and high scanner sound levels. To provide a low sound level experiment, a sparse scanning method was used, characterized by long, 9 sec, periods of scanner silence interspersed with 1 sec echoplanar imaging (EPI) bursts. For the condition with high sound levels, extra EPI gradient modules were inserted in the 9 sec inter-image intervals. Visual, motor, or auditory stimuli were presented in the interval between imaging. It was found that with the addition of gradient sounds, auditory activation was significantly decreased while motor and visual activation were not significantly altered. Other general factors relating to fMRI were also examined, such as experimental duration and fatigue. For example, motion of the subjects during the experiments was found to be related to the time spent in the scanner, rather than to the ambient sound level. PMID- 10371457 TI - Resolution enhancement in single-shot imaging using simultaneous acquisition of spatial harmonics (SMASH). AB - Spatial resolution in single-shot imaging is limited by signal attenuation due to relaxation of transverse magnetization. This effect can be reduced by minimizing acquisition times through the use of short interecho spacings. However, the minimum interecho spacing is constrained by limits on gradient switching rates, radiofrequency (RF) power deposition and RF pulse length. Recently, simultaneous acquisition of spatial harmonics (SMASH) has been introduced as a method to acquire magnetic resonance images at increased speeds using a reduced number of phase-encoding gradient steps by extracting spatial information contained in an RF coil array. In this study, it is shown that SMASH can be used to reduce the effects of relaxation, resulting in single-shot images with increased spatial resolution without increasing imaging time. After a brief theoretical discussion, two strategies to reduce signal attenuation and increase spatial resolution in single-shot imaging are introduced and their performance is evaluated in phantom studies. In vivo single-shot echoplanar imaging (EPI), BURST, and half-Fourier single-shot turbo spin-echo (HASTE) images are then presented demonstrating the practical implementation of these resolution enhancement strategies. Images acquired with SMASH show increased spatial resolution and improved image quality when compared with images obtained with the conventional acquisitions. The general principles presented for imaging with SMASH can also be applied to other partially parallel imaging techniques. PMID- 10371458 TI - QUIPSS II with thin-slice TI1 periodic saturation: a method for improving accuracy of quantitative perfusion imaging using pulsed arterial spin labeling. AB - Quantitative imaging of perfusion using a single subtraction, second version (QUIPSS II) is a pulsed arterial spin labeling (ASL) technique for improving the quantitation of perfusion imaging by minimizing two major systematic errors: the variable transit delay from the distal edge of the tagged region to the imaging slices, and the contamination by intravascular signal from tagged blood that flows through the imaging slices. However, residual errors remain due to incomplete saturation of spins over the slab-shaped tagged region by the QUIPSS II saturation pulse, and spatial mismatch of the distal edge of the saturation and inversion slice profiles. By replacing the original QUIPSS II saturation pulse with a train of thin-slice periodic saturation pulses applied at the distal end of the tagged region, the accuracy of perfusion quantitation is improved. Results of single and multislice studies are reported. PMID- 10371459 TI - Are mono-exponential fits to a few echoes sufficient to determine T2 relaxation for in vivo human brain? AB - T2 relaxation decay curves from in vivo human brain tissue are rarely mono exponential. Partial volume averaging further reduces the chance of mono exponential decay. Moreover, the parameters derived from few-echo mono exponential fits change with the measurement echo times and have the largest possible variance. In this note, multi-exponential fits to 32-echo relaxation decay curves from in vivo human brain are used to design simulations (where the truth is known) to demonstrate the pitfalls of few-echo mono-exponential interpretations. PMID- 10371460 TI - Observing curved flow using RUFIS. AB - The rotating ultra-fast imaging sequence (RUFIS) is used to image spins flowing through the curved portion of a 180 degrees U-tube with a circular cross section and a ratio of inner diameter to curvature diameter of 0.41. A velocity-encoding preparation sequence is used with RUFIS to measure quantitatively the axial velocity distribution for spins under steady-flow conditions. The resulting velocity contours confirm that higher velocity spins migrate toward the outer wall of the curve as the Dean number is increased. PMID- 10371461 TI - Comparison of static and dynamic MRI techniques for the measurement of regional cerebral blood volume. AB - Two different acquisition and processing strategies to determine the regional cerebral blood volume (rCBV) with magnetic resonance imaging (MRI) are compared. The first method is based on the acquisition of the signal time course during a bolus administration of a contrast agent (dynamic method). The second method evaluates signal changes before and after the contrast agent injection (static method), assuming the contrast agent remains primarily intravascular in the brain after the first pass. Both methods were applied to the same data sets, acquired with either echoplanar imaging (EPI, n = 18) or fast low-angle shot (FLASH, n = 28) techniques. A voxel-by-voxel correlation between the static and dynamic method yielded a correlation coefficient of 0.76 +/- 0.06 for the EPI and 0.71 +/ 0.10 for the FLASH measurements. The static method was less sensitive and showed higher standard deviations for rCBV than the dynamic method. With the development of truly intravascular contrast agents, the static perfusion MRI method, which can be performed with higher signal-to-noise ratio and higher spatial resolution, may become an alternative to ultra-fast MRI for measuring rCBV. PMID- 10371462 TI - Magnetic resonance diffusion imaging of the human cervical spinal cord in vivo. AB - Knowledge of water diffusion characteristics within the human spinal cord may provide important information about the structural nature of spinal cord pathology. However, the sensitivity of diffusion imaging methods to motion and the requirement for high in-plane resolution has hitherto restricted study of spinal cord diffusion to excised samples. The first diffusion images of the human cervical spinal cord in vivo are presented using a navigated pulsed-gradient spin echo sequence. Anisotropic diffusion in the cord is demonstrated in agreement with in vitro studies, and further development of the technique for clinical studies is discussed. PMID- 10371463 TI - Quality control for functional magnetic resonance imaging using automated data analysis and Shewhart charting. AB - A data acquisition and analysis protocol for quality control (QC) of functional magnetic resonance imaging studies is presented. Two sets of data are acquired, single-timepoint data for measurement of signal-to-ghost and signal-to-noise ratios, and multiple-timepoint data for measurement of short-term drift. Since manual data analysis can be time consuming and an impediment to regular QC, an automated data processing scheme is presented. The use of automated Shewhart charting is proposed to identify significant changes in each parameter over the long term. The protocol has successfully identified system faults and deteriorations undetected by conventional QC. PMID- 10371464 TI - Comments on "Probabilistic analysis of functional magnetic resonance imaging data". PMID- 10371465 TI - New perspectives in the functional role of GABA(A) channel heterogeneity. AB - Gamma-aminobutyric acid A (GABA(A)) channels responsible for inhibitory synaptic transmission possess a consistent heterogeneity of structure in terms of distinct constitutive subunits. During the past 10 years, considerable progress has been made in understanding the magnitude of this large diversity. Structural requirements for clinically important drugs such as benzodiazepines and barbiturates have been elucidated, and the anatomical distribution in distinct neuronal populations and the developmental profiles of individual subunits have been elucidated with various techniques. However, the relevance of subunit heterogeneity to synaptic transmission is still largely lacking. Recently, substantial progress has been achieved in understanding the crucial role of desensitization as a molecular determinant in defining the duration and frequency responses of inhibitory synaptic transmission. This development, together with a combination of different experimental approaches, including patch-clamp recordings and ultrafast agonist applications in brain slices and mammalian cells expressing recombinant GABA(A) receptor, has begun to shed light on a possible role for subunit composition of synaptic receptors in shaping the physiological characteristics of synaptic transmission. Nowhere else in the central nervous system is the anatomical and developmental profile of GABA receptor heterogeneity as well understood as it is in the cerebellum. This review summarizes advances in the understanding of functional correlates to subunit heterogeneity in the cerebellum relevant for inhibitory synaptic function. PMID- 10371468 TI - Easy access to antifungal agents. PMID- 10371467 TI - Biochemical studies of the structure and function of the N-methyl-D-aspartate subtype of glutamate receptors. AB - The N-methyl-D-aspartate (NMDA) subtype of glutamate receptors plays a key role in synaptic transmission, synaptic plasticity, synaptogenesis, and excitotoxicity in the mammalian central nervous system. The NMDA receptor channel is formed from two gene products from two glutamate receptor subunit families, termed NR1 and NR2. Although the subunit composition of native NMDA receptors is incompletely understood, electrophysiological studies using recombinant receptors suggest that functional NMDA receptors consist of heteromers containing combinations of NR1, which is essential for channel activity, and NR2, which modulates the properties of the channels. The lack of agonists or antagonists selective for a given subunit of NMDA receptors has made it difficult to understand the subunit expression, subunit composition, and posttranslational modification mechanisms of native NMDA receptors. Therefore, most studies on NMDA receptors that examine regional expression and ontogeny have been focused at the level of the mRNAs encoding the different subunits using northern blotting, ribonuclease protection, and in situ hybridization techniques. However, the data from these studies do not provide clear information about the resultant subunit protein. To directly examine the protein product of the NMDA receptor subunit genes, the development of subunit-specific antibodies using peptides and fusion proteins has provided a good approach for localizing, quantifying, and characterizing the receptor subunits in tissues and transfected cell lines, and to study the subunit composition and the functional effects of posttranslational processing of the NMDA subunits, particularly the phosphorylation profiles of NMDA glutamate receptors. PMID- 10371469 TI - Microscopy of the bacterial flora on fresh vaginal smears. PMID- 10371470 TI - Circulating heat shock proteins in women with a history of recurrent vulvovaginitis. AB - OBJECTIVE: Predisposing factors influencing recurrences of bacterial vaginosis (BV) or vaginitis from Candida remain unidentified for most women. As a component of studies to determine host susceptibility factors to genital tract infections in women, we measured expression of the 60-kDa and 70-kDa heat shock proteins (hsp60 and hsp70, respectively) in the circulation of women with or without a history of recurrent BV or candidal vaginitis and with or without a current lower genital tract infection. Heat shock protein expression is associated with a down regulation of pro-inflammatory immune responses that would inhibit microbial infection. METHOD: The investigators measured hsp60 and hsp70, antibodies to these proteins, the pro-inflammatory cytokine tumor necrosis factor-alpha (TNF alpha), and the anti-inflammatory cytokine interleukin-10 (IL-10) in sera by ELISA. The study population consisted of 100 women who attended a gynecology clinic in Campinas, Brazil. Of those, 55 had a history of recurrent vulvovaginitis (RV), while 45 were controls with no such history. Only women who were asymptomatic for at least 1 month were studied. RESULTS: Although all were asymptomatic, clinical and microbiological examination revealed that five of the women with a history of RV and two controls had a current candidal vaginal infection; 16 RV patients and 12 controls had BV; and six RV patients had both BV and candidiasis. Twenty-eight RV patients and 31 controls had no clinical or microbiological detectable vaginal infection. Among the RV patients, hsp60 and hsp70 were more prevalent in those with current BV (40.9% and 50.0%, respectively) or a candidal infection (45.5% and 54.5%) than in women with no current infection (21.4% and 17.9%). In the women with no history of RV, BV was not associated with a high prevalence of hsp60 (8.3%) or hsp70 (8.3%). Interleukin-10 and TNF were not more prevalent in vaginitis patients or controls with a current candidal infection or BV than in uninfected subjects. CONCLUSION: The high prevalence of circulating hsp60 and hsp70 in women with a history of RV and current BV or vaginal candidiasis, but not in women with no history of RV, suggests that differences in heat shock protein induction may be related to susceptibility to recurrent vaginal infections. PMID- 10371471 TI - Single daily dosing of gentamicin: pharmacokinetic comparison of two dosing methodologies for postpartum endometritis. AB - OBJECTIVE: We compared the pharmacokinetics of two methods for dosing gentamicin for the treatment of postpartum endometritis with the goal of achieving adequate peak serum concentrations (>12 mg/L) and prolonged trough levels below 2 mg/L. METHODS: Group-I subjects (n = 5) received intravenous gentamicin, 5 mg/kg per total body weight over 60 min., with a maximum dose of 500 mg. Group-II subjects (n = 17) were dosed intravenously according to the following formula: Dose = desired peak concentration (fixed at 14 mg/L) * (volume of distribution, i.e., 0.35 L/kg) * adjusted body weight (in kilograms). Serum gentamicin levels were obtained 1 hr. and 8-12 hr. after infusion of the second dose. Pharmacokinetic parameters for the subjects in each group were calculated according to standard formulas. RESULTS: Subjects in Group I had significantly higher doses and peak drug concentrations (P < 0.01), while in Group II, 76% of patients had peak levels less than desired (<12 mg/L). Both groups maintained trough levels of <2 mg/L in excess of 12 hr. CONCLUSIONS: Changing to the adjusted body weight formula for Group I, while maintaining a dose between 4 and 5 mg/kg, would reduce excessive peak concentrations. Using a calculated volume of distribution of 0.4 L/kg in Group II would improve peak serum concentrations to the desired levels. Both dosing regimens ensure adequate aminoglycoside pharmacokinetic parameters and avoid the need for monitoring serial serum drug concentrations, provided the expected clinical response is also achieved. While the first dosing formula is simpler to calculate, the second dosing formula allows for more individualized dosing considerations. PMID- 10371472 TI - Evaluation of ofloxacin in the treatment of laparoscopically documented acute pelvic inflammatory disease (salpingitis). AB - OBJECTIVE: To evaluate the safety and efficacy of intravenous and oral ofloxacin monotherapy in the treatment of laparoscopically documented acute pelvic inflammatory disease (PID). METHODS: This study was conducted as an open-label, phase-III, uncontrolled, multicenter study. Patients identified with laparoscopic findings of salpingitis were treated with 400 mg of intravenous ofloxacin every 12 hours followed by 400 mg of oral ofloxacin every 12 hours for 10 to 14 days. Patients were evaluated five times for clinical and microbial efficacy. Since laparoscopy was performed only at admission, pathogens identified laparoscopically were presumed eradicated if they were present on the laparoscopic culture and the patient was clinically cured or improved at final evaluation. RESULTS: Of the 70 patients evaluable for safety (intent-to-treat population), the mean age was 25.6 years. Sixty-one of 70 patients (87%) were cured, one improved, one did not improve, and seven were unevaluable because they discontinued study participation. Fifty-one were evaluable for clinical efficacy: 50 (98%) were cured and one did not improve. Sixteen were evaluable for expanded microbiological efficacy: three had documented Neisseria gonorrhoeae; 12, Chlamydia trachomatis; and one, a mixed infection of both organisms. All cervical, laparoscopic, and endometrial cultured pathogens, including N. gonorrhoeae and C. trachomatis, were eradicated or presumed eradicated at the posttherapy visit. No serious or unexpected adverse events occurred. CONCLUSIONS: Ofloxacin monotherapy was effective and well tolerated in the treatment of laparoscopically proven PID in a geographically diverse population. Future studies are necessary to evaluate long-term outcomes and sequelae of PID treatment with single agent therapy. PMID- 10371466 TI - Structural features of heterotrimeric G-protein-coupled receptors and their modulatory proteins. AB - Over the past 20 years, the general mechanism for signaling through 7 transmembrane helix receptors coupled to GTP hydrolysis has been worked out. Although similar in overall organization, subtype variability and subcellular localization of components have built in considerable signaling specificity. Atomic resolution structures for many of the components have delineated the domain organization of these complex proteins and have given physical form to the idea of subtype specificity. This review describes what is known about the physical structures of the 7-transmembrane helix receptors, the heterotrimeric GTP binding coupling proteins, the adenylate cyclase and phospholipase C effector proteins, and signaling modulatory proteins, such as arrestin, phosducin, recoverin-type myristoyl switch proteins, and the pleckstrin homology domain of G protein receptor kinase-2. These images allow experimenters to contemplate the details of the supramolecular organization of the multiprotein complexes involved in the transmission of signals across the cellular lipid bilayer. PMID- 10371473 TI - Infertility following pelvic inflammatory disease. AB - OBJECTIVE: To assess the frequency of infertility after pelvic inflammatory disease (PID) and factors important in postinfectious tubal damage in an urban population at high risk for sexually transmitted diseases. METHODS: From a cohort of 213 women with PID documented by laparoscopy and/or endometrial biopsy, 58 women (27% of the initial cohort) were interviewed by phone 2 to 9 years after an index episode of PID. Data regarding the initial history, physical examination, microbiology, laparoscopic, and serologic findings, and data concerning interval contraception, subsequent pregnancy, subsequent infection, and chronic pelvic pain were compared among those with and without infertility at follow up. RESULTS: Nineteen (40%) of the 48 women not using contraception were involuntarily infertile after the index episode of PID. Compared with those who had an interval pregnancy, infertile women were older (P = 0.02), more likely to have a history of infertility prior to the index episode of PID (P = 0.001), and were more likely to have occluded or partially occluded fallopian tubes (P = 0.03), peritubal adhesions (P = 0.007), or perihepatic adhesions (P = 0.02) seen by laparoscopy performed during the index episode. Surprisingly, recovery of Chlamydia trachomatis was negatively related to infertility (P = 0.001), although a similar proportion of both groups had chlamydia immunoglobulin M antibody (40% vs. 31%). Chlamydia heat shock protein was weakly related to infertility (P = 0.08). The isolation of Neisseria gonorrhoeae was not significantly different between groups (53% vs. 57%). CONCLUSIONS: The high rate of postinfection infertility found was probably related to a combination of tubal damage before and during the index episode of PID. Prevention of recurrent PID and better understanding of the pathophysiology of postinfection tubal damage (which may differ between chlamydia and gonorrhea) is needed to develop more effective strategies to reduce permanent tubal damage. PMID- 10371474 TI - Prophylactic cefazolin in amnioinfusions administered for meconium-stained amniotic fluid. AB - OBJECTIVE: To determine if amnioinfusion with an antibiotic solution decreased the rate of clinical chorioamnionitis and puerperal endometritis in patients with meconium-stained amniotic fluid. METHODS: Patients in labor at 36 weeks of gestation or greater with singleton pregnancies and meconium-stained amniotic fluid were randomized to receive either cefazolin, 1 g/1,000 mL, of normal saline (n = 90) or normal saline (n = 93) amnioinfusion. Rates of clinically diagnosed chorioamnionitis and endometritis and of suspected and culture-proven neonatal infection were determined. RESULTS: Between the study and control groups, the incidences of clinical chorioamnionitis (7.8% vs. 8.6%), endometritis (2.4% vs. 3.5%), aggregate intrauterine infection (10.0% vs. 11.8%), suspected neonatal infection (17.8% vs. 21.5%), and proven neonatal infection (0.0% vs. 2.2%) were not significantly different. CONCLUSIONS: Prophylactic use of cefazolin in amnioinfusions did not significantly reduce rates of maternal or neonatal infection in patients with meconium-stained amniotic fluid. PMID- 10371476 TI - Ligneous cellulitis following spontaneous vaginal delivery. PMID- 10371475 TI - A matched prospective study of human immunodeficiency virus serostatus, human papillomavirus DNA, and cervical lesions detected by cytology and colposcopy. AB - OBJECTIVE: To compare the prevalence and type of human papillomavirus (HPV) infections in the genital tract of human-immunodeficiency-virus- (HIV) seropositive and -seronegative women matched for cytology and to examine prospectively the relationship of HPV DNA, colposcopic findings and cervical squamous intraepithelial lesions (SIL) in these matched seropositive and seronegative cohorts. METHODS: A matched prospective study of HIV-seropositive and -seronegative women undergoing cytologic screening, colposcopy, and testing for HPV DNA and other infections at each visit. RESULTS: Twenty-three HIV seropositive women were matched with 23 seronegative women by cervical cytology reading, lifetime number of sexual partners, age, and follow-up length. Fourteen pairs of these women had follow-up visits every 4 months, for 56 and 53 total visits in seropositive and seronegative women, respectively. After matching, the groups had a similar overall prevalence of HPV DNA and of HPV oncogenic (high risk) types at baseline. On follow up, HIV-seropositive women were more likely than seronegative women to develop SIL (38% vs. 10%), less likely to have negative cytology (34% vs. 60%, overall P = 0.03), more visits with HPV DNA detected (68% vs. 40% P = 0.04), and more visits with multiple HPV DNA types detected (18% vs. 0%, P = 0.02). Colposcopic lesions in the seropositive women were more likely to have sharp borders or mosaicism or to be thick white (P = 0.009). CONCLUSIONS: After matching for baseline Papanicolaou smear readings, these data suggest that over time seropositive women have more visits that yield abnormal cytology, more persistent HPV DNA detection, and more colposcopic abnormalities than seronegative women. PMID- 10371477 TI - The role of bacterial vaginosis in infection after major gynecologic surgery. AB - PURPOSE: Previous studies have reported an association between bacterial vaginosis (BV) and postoperative fever and infection. This prospective study investigated whether the intermediate or definite stages of BV are risk factors for postoperative infection after major gynecologic surgery. METHODS: Vaginal cultures were obtained preoperatively from 175 women undergoing gynecologic surgery. The diagnostic criteria for BV were based on Nugent's standardized method of Gram stain interpretation. Postoperative fever was defined as at least one temperature equal to 101.0 degrees F or greater, or two or more temperatures more than 6 hours apart equal to 100.4 degrees F or greater. RESULTS: Thirty-six percent of the positive-BV group developed a postoperative fever, compared with 20% of the Lactobacillus-predominant group and 12% of the intermediate-BV group (P = 0.017). The differences between the positive-BV group and the Lactobacillus predominant group, and between the positive-BV group and the intermediate-BV group, with respect to postoperative fever, were statistically significant (P = 0.045 and P = 0.007, respectively). The difference between the intermediate-BV group and the Lactobacillus-predominant group was not statistically significant (P = 0.28). CONCLUSIONS: Although the association between BV and postoperative febrile morbidity could be a spurious result of confounding with other variables, it may be prudent for the surgeon to identify patients with BV and treat them preoperatively. PMID- 10371478 TI - A higher standard for human research. PMID- 10371479 TI - Protecting Japan from influenza. PMID- 10371480 TI - Cytokine modulation of HIV-1 chemokine receptor expression. PMID- 10371481 TI - Keeping an eye on ICSI. PMID- 10371482 TI - Deal creates lead cloning company. PMID- 10371483 TI - World Bank evaluates the economics of tobacco. PMID- 10371484 TI - Japan considers the achievements and pitfalls of its five-year plan. PMID- 10371485 TI - Australian budget pleases most researchers. PMID- 10371486 TI - WMA to revise medical research guidelines. World Medical Association. PMID- 10371487 TI - BMS offers some relief for Africa's AIDS crisis. Bristol Myers Squibb. PMID- 10371488 TI - The NOD mouse model of type 1 diabetes: as good as it gets? PMID- 10371489 TI - SAP-less chromatin triggers systemic lupus erythematosus. PMID- 10371490 TI - HIV-1 and HAART: a time to cure, a time to kill. PMID- 10371491 TI - Latent reservoirs of HIV infection: flushing with IL-2? PMID- 10371492 TI - Is an HIV vaccine possible? PMID- 10371493 TI - Big deal about a little insulin. PMID- 10371494 TI - Graft tolerance: a duel of two signals. PMID- 10371495 TI - Cartilage to bone--angiogenesis leads the way. PMID- 10371496 TI - Blasting away leukemia. PMID- 10371497 TI - Victory at C. PMID- 10371498 TI - COX-2 inhibitors--is there cause for concern? PMID- 10371499 TI - VEGF couples hypertrophic cartilage remodeling, ossification and angiogenesis during endochondral bone formation. AB - Hypertrophic chondrocytes in the epiphyseal growth plate express the angiogenic protein vascular endothelial growth factor (VEGF). To determine the role of VEGF in endochondral bone formation, we inactivated this factor through the systemic administration of a soluble receptor chimeric protein (Flt-(1-3)-IgG) to 24-day old mice. Blood vessel invasion was almost completely suppressed, concomitant with impaired trabecular bone formation and expansion of hypertrophic chondrocyte zone. Recruitment and/or differentiation of chondroclasts, which express gelatinase B/matrix metalloproteinase-9, and resorption of terminal chondrocytes decreased. Although proliferation, differentiation and maturation of chondrocytes were apparently normal, resorption was inhibited. Cessation of the anti-VEGF treatment was followed by capillary invasion, restoration of bone growth, resorption of the hypertrophic cartilage and normalization of the growth plate architecture. These findings indicate that VEGF-mediated capillary invasion is an essential signal that regulates growth plate morphogenesis and triggers cartilage remodeling. Thus, VEGF is an essential coordinator of chondrocyte death, chondroclast function, extracellular matrix remodeling, angiogenesis and bone formation in the growth plate. PMID- 10371500 TI - Production of atypical measles in rhesus macaques: evidence for disease mediated by immune complex formation and eosinophils in the presence of fusion-inhibiting antibody. AB - The severe disease atypical measles occurred when individuals immunized with a poorly protective inactivated vaccine contracted measles, and was postulated to be due to a lack of fusion-inhibiting antibodies. Here, rhesus macaques immunized with formalin-inactivated measles vaccine developed transient neutralizing and fusion-inhibiting antibodies, but no cytotoxic T-cell response. Subsequent infection with measles virus caused an atypical rash and pneumonitis, accompanied by immune complex deposition and an increase in eosinophils. Fusion-inhibiting antibody appeared earlier in these monkeys than in non-immunized monkeys. These data indicate that atypical measles results from previous priming for a nonprotective type 2 CD4 T-cell response rather than from lack of functional antibody against the fusion protein. PMID- 10371501 TI - Inhibition of HIV replication by dominant negative mutants of Sam68, a functional homolog of HIV-1 Rev. AB - The HIV-1 Rev protein facilitates the nuclear export of mRNA containing the Rev response element (RRE) through binding to the export receptor CRM-1. Here we show that a cellular nuclear protein, Sam68 (Src-associated protein in mitosis), specifically interacts with RRE and can partially substitute for as well as synergize with Rev in RRE-mediated gene expression and virus replication. Differential sensitivity to leptomycin B, an inhibitor of CRM-1, indicates that the export pathways mediated by Rev and Sam68 are distinct. C-terminally deleted mutants of Sam68 inhibited the transactivation of RRE-mediated expression by both wild-type Sam68 and Rev. They were retained in the cytoplasm and impeded the nuclear localization of Rev in co-expressed cells. These mutants also inhibited wild-type HIV-1 replication to the same extent as the RevM10 mutant, and may be useful as anti-viral agents in the treatment of AIDS. PMID- 10371502 TI - Control of SHIV-89.6P-infection of cynomolgus monkeys by HIV-1 Tat protein vaccine. AB - Vaccine strategies aimed at blocking virus entry have so far failed to induce protection against heterologous viruses. Thus, the control of viral infection and the block of disease onset may represent a more achievable goal of human immunodeficiency virus (HIV) vaccine strategies. Here we show that vaccination of cynomolgus monkeys with a biologically active HIV-1 Tat protein is safe, elicits a broad (humoral and cellular) specific immune response and reduces infection with the highly pathogenic simian-human immunodeficiency virus (SHIV)-89.6P to undetectable levels, preventing the CD4+ T-cell decrease. These results may provide new opportunities for the development of a vaccine against AIDS. PMID- 10371503 TI - Effect of interleukin-2 on the pool of latently infected, resting CD4+ T cells in HIV-1-infected patients receiving highly active anti-retroviral therapy. AB - The size of the pool of resting CD4+ T cells containing replication-competent HIV in the blood of patients receiving intermittent interleukin (IL)-2 plus highly active anti-retroviral therapy (HAART) was significantly lower than that of patients receiving HAART alone. Virus could not be isolated from the peripheral blood CD4+ T cells in three patients receiving IL-2 plus HAART, despite the fact that large numbers of resting CD4+ T cells were cultured. Lymph node biopsies were done in two of these three patients and virus could not be isolated. These results indicate that the intermittent administration of IL-2 with continuous HAART may lead to a substantial reduction in the pool of resting CD4+ T cells that contain replication-competent HIV. PMID- 10371504 TI - The intrinsic factor-vitamin B12 receptor, cubilin, is a high-affinity apolipoprotein A-I receptor facilitating endocytosis of high-density lipoprotein. AB - Cubilin is the intestinal receptor for the endocytosis of intrinsic factor vitamin B12. However, several lines of evidence, including a high expression in kidney and yolk sac, indicate it may have additional functions. We isolated apolipoprotein A-I (apoA-I), the main protein of high-density lipoprotein (HDL), using cubilin affinity chromatography. Surface plasmon resonance analysis demonstrated a high-affinity binding of apoA-I and HDL to cubilin, and cubilin expressing yolk sac cells showed efficient 125I-HDL endocytosis that could be inhibited by IgG antibodies against apoA-I and cubilin. The physiological relevance of the cubilin-apoA-I interaction was further emphasized by urinary apoA-I loss in some known cases of functional cubilin deficiency. Therefore, cubilin is a receptor in epithelial apoA-I/HDL metabolism. PMID- 10371505 TI - Extracellular matrix proteins protect small cell lung cancer cells against apoptosis: a mechanism for small cell lung cancer growth and drug resistance in vivo. AB - Resistance to chemotherapy is a principal problem in the treatment of small cell lung cancer (SCLC). We show here that SCLC is surrounded by an extensive stroma of extracellular matrix (ECM) at both primary and metastatic sites. Adhesion of SCLC cells to ECM enhances tumorigenicity and confers resistance to chemotherapeutic agents as a result of beta1 integrin-stimulated tyrosine kinase activation suppressing chemotherapy-induced apoptosis. SCLC may create a specialized microenvironment, and the survival of cells bound to ECM could explain the partial responses and local recurrence of SCLC often seen clinically after chemotherapy. Strategies based on blocking beta1 integrin-mediated survival signals may represent a new therapeutic approach to improve the response to chemotherapy in SCLC. PMID- 10371506 TI - Ligation of the CD44 adhesion molecule reverses blockage of differentiation in human acute myeloid leukemia. AB - Blockage in myeloid differentiation characterizes acute myeloid leukemia (AML); the stage of the blockage defines distinct AML subtypes (AML1/2 to AML5). Differentiation therapy in AML has recently raised interest because the survival of AML3 patients has been greatly improved using the differentiating agent retinoic acid. However, this molecule is ineffective in other AML subtypes. The CD44 surface antigen, on leukemic blasts from most AML patients, is involved in myeloid differentiation. Here, we report that ligation of CD44 with specific anti CD44 monoclonal antibodies or with hyaluronan, its natural ligand, can reverse myeloid differentiation blockage in AML1/2 to AML5 subtypes. The differentiation of AML blasts was evidenced by the ability to produce oxidative bursts, the expression of lineage antigens and cytological modifications, all specific to normal differentiated myeloid cells. These results indicate new possibilities for the development of CD44-targeted differentiation therapy in the AML1/2 to AML5 subtypes. PMID- 10371507 TI - Characterization of circulating T cells specific for tumor-associated antigens in melanoma patients. AB - We identified circulating CD8+ T-cell populations specific for the tumor associated antigens (TAAs) MART-1 (27-35) or tyrosinase (368-376) in six of eleven patients with metastatic melanoma using peptide/HLA-A*0201 tetramers. These TAA-specific populations were of two phenotypically distinct types: one, typical for memory/effector T cells; the other, a previously undescribed phenotype expressing both naive and effector cell markers. This latter type represented more than 2% of the total CD8+ T cells in one patient, permitting detailed phenotypic and functional analysis. Although these cells have many of the hallmarks of effector T cells, they were functionally unresponsive, unable to directly lyse melanoma target cells or produce cytokines in response to mitogens. In contrast, CD8+ T cells from the same patient were able to lyse EBV-pulsed target cells and showed robust allogeneic responses. Thus, the clonally expanded TAA-specific population seems to have been selectively rendered anergic in vivo. Peptide stimulation of the TAA-specific T-cell populations in other patients failed to induce substantial upregulation of CD69 expression, indicating that these cells may also have functional defects, leading to blunted activation responses. These data demonstrate that systemic TAA-specific T-cell responses can develop de novo in cancer patients, but that antigen-specific unresponsiveness may explain why such cells are unable to control tumor growth. PMID- 10371508 TI - Treatment with humanized monoclonal antibody against CD154 prevents acute renal allograft rejection in nonhuman primates. AB - CD154 is the ligand for the receptor CD40. This ligand-receptor pair mediates endothelial and antigen-presenting cell activation, and facilitates the interaction of these cells with T cells and platelets. We demonstrate here that administration of a CD154-specific monoclonal antibody (hu5C8) allows for renal allotransplantation in outbred, MHC-mismatched rhesus monkeys without acute rejection. The effect persisted for more than 10 months after therapy termination, and no additional drug was required to achieve extended graft survival. Indeed, the use of tacrolimus or chronic steroids seemed to antagonize the anti-rejection effect. Monkeys treated with antibody against CD154 remained healthy during and after therapy. The mechanism of action does not require global depletion of T or B cells. Long-term survivors lost their mixed lymphocyte reactivity in a donor-specific manner, but still formed donor-specific antibody and generated T cells that infiltrated the grafted organ without any obvious effect on graft function. Thus, therapy with antibody against CD154 is a promising agent for clinical use in human allotransplantation. PMID- 10371509 TI - Serum amyloid P component controls chromatin degradation and prevents antinuclear autoimmunity. AB - Serum amyloid P component (SAP), a highly conserved plasma protein named for its universal presence in amyloid deposits, is the single normal circulating protein that shows specific calcium-dependent binding to DNA and chromatin in physiological conditions. The avid binding of SAP displaces H1-type histones and thereby solubilizes native long chromatin, which is otherwise profoundly insoluble at the physiological ionic strength of extracellular fluids. Furthermore, SAP binds in vivo both to apoptotic cells, the surface blebs of which bear chromatin fragments, and to nuclear debris released by necrosis. SAP may therefore participate in handling of chromatin exposed by cell death. Here we show that mice with targeted deletion of the SAP gene spontaneously develop antinuclear autoimmunity and severe glomerulonephritis, a phenotype resembling human systemic lupus erythematosus, a serious autoimmune disease. The SAP-/- mice also have enhanced anti-DNA responses to immunization with extrinsic chromatin, and we demonstrate that degradation of long chromatin is retarded in the presence of SAP both in vitro and in vivo. These findings indicate that SAP has an important physiological role, inhibiting the formation of pathogenic autoantibodies against chromatin and DNA, probably by binding to chromatin and regulating its degradation. PMID- 10371510 TI - Inducible cyclooxygenase may have anti-inflammatory properties. AB - Cyclooxygenase (COX) has two isoforms. Generally, COX 1 is constitutively expressed in most tissues, where it maintains physiological processes; inducible COX 2 is considered a pro-inflammatory enzyme and a chief target for the treatment of inflammatory diseases. Here we present evidence that COX 2 may have anti-inflammatory properties. In carrageenin-induced pleurisy in rats, the predominant cells at 2 hours are polymorphonuclear leucocytes, whereas mononuclear cells dominate from 24 hours until resolution at 48 hours. In this model, COX 2 protein expression peaked initially at 2 hours, associated with maximal prostaglandin E2 synthesis. However, at 48 hours there was a second increase in COX 2 expression, 350% greater than that at 2 hours. Paradoxically, this coincided with inflammatory resolution and was associated with minimal prostaglandin E2 synthesis. In contrast, levels of prostaglandin D2, and 15deoxy delta(12-14)prostaglandin J2 were high at 2 hours, decreased as inflammation increased, but were increased again at 48 hours. The selective COX 2 inhibitor NS 398 and the dual COX 1/COX 2 inhibitor indomethacin inhibited inflammation at 2 hours but significantly exacerbated inflammation at 48 hours. This exacerbation was associated with reduced exudate prostaglandin D2 and 15deoxy delta(12 14)prostaglandin J2 concentrations, and was reversed by replacement of these prostaglandins. Thus, COX 2 may be pro-inflammatory during the early phase of a carrageenin-induced pleurisy, dominated by polymorphonuclear leucocytes, but may aid resolution at the later, mononuclear cell-dominated phase by generating an alternative set of anti-inflammatory prostaglandins. PMID- 10371511 TI - Intra-articularly localized bacterial DNA containing CpG motifs induces arthritis. AB - Unmethylated CpG motifs are often found in bacterial DNA, and exert immunostimulatory effects on hematopoietic cells. Bacteria produce severe joint inflammation in septic and reactive arthritides; bacterial DNA may be involved in this process. We injected bacterial DNA originating from Escherichia coli and Staphylococcus aureus and oligonucleotides containing CpG directly into the knee joints of mice of different strains. Arthritis was seen by histopathology within 2 hours and lasted for at least 14 days. Unmethylated CpG motifs were responsible for this induction of arthritis, as oligonucleotides containing these motifs produced the arthritis. The arthritis was characterized by an influx of monocytic, Mac-1+ cells and by a lack of T lymphocytes. Depletion of monocytes resulted in abrogation of the synovial inflammation. Tumor necrosis factor (TNF) alpha, a cytokine produced by cells of the monocyte/macrophage lineage, is an important mediator of this disease, as expression of mRNA for TNF-alpha was evident in the inflamed joints, and the CpG-mediated inflammation was abrogated in mice genetically unable to produce this cytokine. These findings demonstrate that bacterial DNA containing unmethylated CpG motifs induces arthritis, and indicate an important pathogenic role for bacterial DNA in septic arthritis. PMID- 10371512 TI - New delivery system for plasmid DNA in vivo using atelocollagen as a carrier material: the Minipellet. PMID- 10371513 TI - Interferon beta and the cytokine trail: where are we going? PMID- 10371514 TI - The Webster's dictionary: neurologists on the Internet. PMID- 10371515 TI - Waking up to the importance of sleep in neurologic disorders. PMID- 10371516 TI - New reasons for early use of MRI in stroke. PMID- 10371517 TI - The relationship of MS to physical trauma and psychological stress: report of the Therapeutics and Technology Assessment Subcommittee of the American Academy of Neurology. PMID- 10371518 TI - Intravenous heparin for acute stroke: what can we learn from the megatrials? AB - Recently published large clinical trials of heparin and aspirin in acute stroke the International Stroke Trial, Chinese Acute Stroke Trial, and Trial of ORG 10172 in Acute Stroke Treatment--fail to show a net benefit from heparin. None of these trials used i.v., dose-adjusted, unfractionated heparin as generally employed in the United States. However, the control groups in these trials provide data on acute stroke recurrence in large numbers of patients, and these stroke recurrence rates can be used to establish an upper limit for the potential efficacy of antithrombotic therapy. The rates of recurrent ischemic stroke in the control groups of these trials were low, ranging from 0.6 to 2.2% per week. The low rates of recurrent stroke observed in these groups, coupled with the morbidity and mortality associated with i.v. heparin in this patient population, argue against routine use of i.v. heparin in the acute stroke period. PMID- 10371519 TI - Applications of diffusion-perfusion magnetic resonance imaging in acute ischemic stroke. AB - Diffusion-weighted imaging (DWI) and perfusion imaging (PI) are two new magnetic resonance technologies that are becoming increasingly available for evaluation of acute ischemic stroke patients. DWI provides information about the location of acute focal ischemic brain injury at early time points and PI can document the presence of disturbances in microcirculatory perfusion. DWI and PI are now being used in clinical practice and in clinical trials of potential acute stroke therapies to assess their utility. In the future, DWI and PI may aid in the development of effective acute stroke therapies and help identify which stroke patients are most likely to benefit from specific agents. PMID- 10371521 TI - Interferon beta-1b treatment modulates TNFalpha and IFNgamma spontaneous gene expression in MS. AB - BACKGROUND: Interferon beta (IFNbeta) lessens the overall frequency of acute attacks in patients with the relapsing-remitting form of multiple sclerosis (RRMS). IFNbeta may act by decreasing the synthesis of inflammatory cytokines. OBJECTIVES: To determine whether IFNbeta-1b treatment had an initial and sustained effect on the in vivo synthesis and secretion of tumor necrosis factor alpha (TNFalpha) and IFNgamma. METHODS: A highly sensitive reverse-transcriptase PCR technique was used to measure baseline levels of mRNA in freshly isolated cells from patients before therapy and at 3, 6, and 12 months of treatment. Also, protein concentration was measured in serum and in culture supernatants from mitogen-stimulated cells. The authors studied 16 patients, of whom 11 did not have clinical exacerbations, whereas 5 had one clinical relapse each during the study. RESULTS: Mean values of TNFalpha mRNA levels in the 11 stable patients decreased significantly at 3 and 6 months of treatment in comparison with initial data. After 6 months of therapy, IFNbeta-1b downmodulated TNFalpha transcripts in the 5 patients who experienced relapse. In this group of patients, TNFalpha levels rose sharply to reach pretreated values at 1 year of IFNbeta-1b treatment. At the beginning of therapy, 6 patients had high concentrations of serum TNFalpha, which decreased to normal values following IFNbeta-1b therapy. IFNgamma mRNA expression also diminished after 6 and 12 months of IFNbeta-1b therapy in the group of stable patients, whereas nonrelevant variations were observed in patients who had one relapse. Initially, patients' peripheral mononuclear cells secreted diminished amounts of TNFalpha and IFNgamma on PHA + PMA mitogen stimulation in comparison with normal control subjects. After 1 year of therapy, IFNbeta-1b restored the normal production of TNFalpha, whereas therapy did not restore IFNgamma secretion to control values. CONCLUSION: IFNbeta-1b decreases the spontaneous expression of two proinflammatory cytokines. PMID- 10371520 TI - A subtype of sporadic prion disease mimicking fatal familial insomnia. AB - OBJECTIVE: To establish a variant of sporadic prion disease as the sporadic form of fatal familial insomnia (FFI). BACKGROUND: FFI is a recently described prion disease characterized clinically by severe sleep impairment, dysautonomia, and motor signs, and pathologically by atrophy of thalamic nuclei, especially the medial dorsal and anterior ventral, and of the inferior olive. FFI is linked to the D178N mutation coupled with the methionine codon at position 129 in the prion protein gene (PRNP). It is also identified by the properties of the abnormal prion protein (PrP(Sc)), which has the relative molecular mass of 19 kDa, corresponding to the so-called type 2, and a marked underrepresentation of the unglycosylated form relative to the diglycosylated and monoglycosylated forms. METHODS: Clinical, pathologic, PrP(Sc), and PRNP data from 5 subjects with a sporadic prion disease phenotypically similar to FFI were collected and analyzed. RESULTS: All 5 subjects had a disease clinically similar and histopathologically virtually identical to FFI. PrP(Sc) type 2 was present in all subjects in amount and distribution similar to those of FFI. However, the PrP(Sc) did not show the striking underrepresentation of the unglycosylated isoform of the protein that is characteristic of FFI. Moreover, none of the subjects had the D178N PRNP mutation but all were homozygous for methionine at codon 129. CONCLUSION: This condition is likely to represent the sporadic form of FFI and the term "sporadic fatal insomnia" is proposed. PMID- 10371522 TI - Immunoglobulin G Fc-receptor (FcgammaR) IIA and IIIB polymorphisms related to disability in MS. AB - OBJECTIVE: MS is immunologically mediated in genetically susceptible individuals. Receptors for the Fc fragment of immunoglobulin G (IgG) (FcgammaR) link the humoral and cellular immune responses by targeting immune complexes to effector cells. Different FcgammaR show variability in their distribution, strength, and capacity of binding different IgG subclasses. METHODS: To investigate the role of FcgammaR in MS, 136 MS patients and 96 matched controls were genotyped for FcgammaRIIA and FcgammaRIIIB gene polymorphisms; the results were correlated to disease susceptibility and severity measured by the Expanded Disability Status Scale (EDSS). RESULTS: The allele frequencies of the FcgammaRIIA and FcgammaRIIIB did not differ significantly between the MS patients and the controls. Patients homozygous for the FcgammaRIIIB neutrophil antigen (NA) 1 allele had a significantly more benign course of MS than patients heterozygous or homozygous for the FcgammaRIIIB NA2 allele. Patients homozygous for the FcgammaRIIA histidine (H) allele also had a more benign course of MS than patients heterozygous or homozygous for the FcgammaRIIA arginine (R) allele. CONCLUSION: The results implicate FcgammaRIIIB and to a lesser extent FcgammaRIIA as disease modifying genes in MS. FcgammaRIIIB NA1/NA1 and FcgammaRIIA H/H bind more efficient IgG1/IgG3 and IgG2 subclasses, respectively, than FcgammaRIIIB NA2/NA2 and FcgammaRIIA R/R. A more effective processing of circulating immune complexes may be one mechanism for better clinical outcome in MS. PMID- 10371523 TI - Respiratory disturbances during sleep in syringomyelia and syringobulbia. AB - OBJECTIVE: To determine the frequency and types of abnormalities of respiratory control during sleep in syringomyelia and syringobulbia and to provide a basis to predict patients at risk of sudden death. METHODS: Thirty patients (15 male and 15 female; mean age 39.0 +/- 12.6 years) with communicating syringomyelia were divided into two groups: those with evidence of syringobulbia (17 patients) and those without compromise of the medulla or syringomyelia (13 patients). Patients were studied with pulmonary function studies and polysomnography. Respiratory center sensitivity to CO2 (rebreathing technique) was measured in 9 patients. RESULTS: Severely affected patients had mild-to-moderate restriction and individual patients had bilateral diaphragmatic or vocal cord palsy, abnormal respiratory rhythm, prolonged inspiratory time, or an abnormal respiratory response to CO2. Very prolonged central, obstructive, and mixed sleep apneas with low O2 saturation values and a fixed heart rate were recorded in most patients with syringobulbia. Five patients developed severe respiratory complications and died during a follow-up period of 10 years. Respiratory abnormalities failed to correlate with syrinx size. CONCLUSIONS: Severe abnormalities in respiratory rhythm generation during sleep occur in patients with syringobulbia. The respiratory disturbances are not due to muscle weakness and they are not correlated with the size of the cavity. The combination of dysphagia and dysphonia in patients with longstanding syringomyelia and syringobulbia predicted likelihood of respiratory disturbances during sleep. PMID- 10371526 TI - Surgical and endovascular treatment of unruptured cerebral aneurysms at university hospitals. AB - OBJECTIVE: To compare complications of surgical clipping and coil embolization in the treatment of unruptured aneurysms. BACKGROUND: Surgical clipping has been the preferred treatment for unruptured cerebral aneurysms but endovascular coil embolization is an increasingly employed alternative. No direct comparisons of the techniques are available to guide clinical decision making. METHODS: We performed a cohort study of patients treated for unruptured cerebral aneurysms at 60 university hospitals from January 1994 through June 1997 using the University HealthSystem Consortium database. The database was validated by chart review from one of the participant universities. The main outcome measures were in-hospital mortality and adverse outcomes, defined as in-hospital deaths and discharges to nursing homes or rehabilitation hospitals. RESULTS: The primary treatment modality was surgical in 2,357 cases and endovascular in 255 cases. Adverse outcomes were significantly more common in surgical cases (18.5%) compared to endovascular cases (10.6%) (p = 0.002), and the difference was not altered after adjusting for age, sex, race, transfer admissions, emergency room admissions, and year of treatment (odds ratio [OR] 2.1, 95% confidence interval [CI] 1.4 to 3.3; p = 0.001). In-hospital mortality was also increased in surgical cases (2.3% versus 0.4%; p = 0.039), but the difference was not significant in the multivariable model (OR 6.3, 95% CI 0.9 to 46.1; p = 0.07). Length of stay and hospital charges were significantly greater for surgical cases (p < 0.0001 for each), and these differences were not affected by risk adjustment. CONCLUSION: Endovascular coil embolization resulted in fewer adverse outcomes than surgery for unruptured cerebral aneurysms treated at the university hospitals studied. Although these results should be seen as preliminary, the magnitude of difference and current predominance of surgery appear to justify a randomized trial. PMID- 10371525 TI - Evaluation of early reperfusion and i.v. tPA therapy using diffusion- and perfusion-weighted MRI. AB - OBJECTIVE: To characterize the effects of recombinant tissue plasminogen activator (rt-PA) therapy and early reperfusion on diffusion-weighted (DWI) and perfusion-weighted imaging (PWI) changes observed following acute ischemic injury. METHODS: Twelve patients were evaluated prospectively using echo planar DWI and bolus tracking PWI. Six patients received i.v. rt-PA 0.9 mg/kg and were compared with six patients who did not. Patients receiving rt-PA were initially imaged (T1) 3 to 5 hours postictus (mean, 4 hours 20 minutes) whereas those not treated with tissue plasminogen activator (tPA) were imaged 4 to 7 hours postictus (mean, 5 hours, 25 minutes). Follow-up imaging was performed 3 to 6 hours (T2), 24 to 36 hours (T3), 5 to 7 days (T4), and 30 days (T5) after the first scan in all patients. Lesion volumes were measured on both DWI and time-to peak maps constructed from PW images. RESULTS: PWI was performed successfully at T1 and T3 in 11 of 12 patients. In the group that received i.v. tPA, initial PWI volumes were less than DWI volumes in five of six patients (83%), whereas only one of five patients (20%) not receiving tPA had PWI < DWI volume (p = 0.08). PWI normalized by 24 to 36 hours (T3) in 6 of 11 patients (early reperfusers), with 5 of 6 of these early reperfusers having received tPA. The aggregate apparent diffusion coefficient (ADC) values for the early reperfusers were consistently higher at T2 (p = 0.04), T3 (p = 0.002), and T4 (p = 0.0005). Five of six patients with early reperfusion demonstrated regions of elevated ADC within the ischemic zone (mean ipsilateral ADC/contralateral ADC, 1.46 +/- 0.19) by 24 to 36 hours, whereas none of the nonearly reperfusers showed these regions of elevated ADC (p = 0.015). CONCLUSION: Early reperfusion is seen more frequently with i.v. tPA therapy. In addition, the study showed that ADC may undergo early increases that are tied closely to reperfusion, and marked ADC heterogeneity may exist within the same lesion. Early reperfusion is seen more frequently with i.v. tPA therapy. PMID- 10371524 TI - Normal diffusion-weighted MRI during stroke-like deficits. AB - BACKGROUND: Diffusion-weighted MRI (DWI) represents a major advance in the early diagnosis of acute ischemic stroke. When abnormal in patients with stroke-like deficit, DWI usually establishes the presence and location of ischemic brain injury. However, this is not always the case. OBJECTIVE: To investigate patients with stroke-like deficits occurring without DWI abnormalities in brain regions clinically suspected to be responsible. METHODS: We identified 27 of 782 consecutive patients scanned when stroke-like neurologic deficits were still present and who had normal DWI in the brain region(s) clinically implicated. Based on all the clinical and radiologic data, we attempted to arrive at a pathophysiologic diagnosis in each. RESULTS: Best final diagnosis was a stroke mimic in 37% and a cerebral ischemic event in 63%. Stroke mimics (10 patients) included migraine, seizures, functional disorder, transient global amnesia, and brain tumor. The remaining patients were considered to have had cerebral ischemic events: lacunar syndrome (7 patients; 3 with infarcts demonstrated subsequently) and hemispheric cortical syndrome (10 patients; 5 with TIA, 2 with prolonged reversible deficits, 3 with infarction on follow-up imaging). In each of the latter three patients, the regions destined to infarct showed decreased perfusion on the initial hemodynamically weighted MRI (HWI). CONCLUSIONS: Normal DWI in patients with stroke-like deficits should stimulate a search for nonischemic cause of symptoms. However, more than one-half of such patients have an ischemic cause as the best clinical diagnosis. Small brainstem lacunar infarctions may escape detection. Concomitant HWI can identify some patients with brain ischemia that is symptomatic but not yet to the stage of causing DWI abnormality. PMID- 10371527 TI - Which unruptured cerebral aneurysms should be treated? A cost-utility analysis. AB - OBJECTIVE: To determine which unruptured cerebral aneurysms should be treated considering the risks. benefits, and costs. BACKGROUND: Asymptomatic unruptured cerebral aneurysms are commonly treated by surgical clipping or endovascular coil embolization to prevent subarachnoid hemorrhage (SAH). METHODS: We performed a cost-utility analysis comparing surgical clipping and endovascular coil embolization with no treatment for unruptured aneurysms. Eight clinical scenarios were defined based on aneurysm size, symptoms, and history of SAH from a different aneurysm. Health outcomes of a hypothetical cohort of 50-year-old women were modeled over the projected lifetime of the cohort. Costs were assessed from the societal perspective. We compared net quality-adjusted life years (QALYs) and cost per QALY of each therapy to no treatment. RESULTS: For an asymptomatic unruptured aneurysm less than 10 mm in diameter in patients with no history of SAH from a different aneurysm, both procedures resulted in a net loss in QALYs, and confidence intervals (CI) were not compatible with a benefit from treatment (clipping, loss of 1.6 QALY [95% CI 1.1 to 2.1]; coiling, loss of 0.6 QALY [95% CI 0.2 to 0.8]). For larger aneurysms (> or = 10 mm), those producing symptoms by compressing neighboring nerves and brain structures, or in patients with a history of SAH from a different aneurysm, treatment was cost-effective. Coiling appeared more effective and cost-effective than clipping but these differences depended on relatively uncertain model parameters. CONCLUSIONS: Treatment of small, asymptomatic, unruptured cerebral aneurysms in patients without a history of SAH worsens clinical outcomes, and thus is neither effective nor cost effective. For aneurysms that are > or = 10 mm or symptomatic, or in patients with a history of SAH, treatment appears to be cost-effective. PMID- 10371528 TI - High prevalence of CACNA1A truncations and broader clinical spectrum in episodic ataxia type 2. AB - OBJECTIVE: To characterize the nature of CACNA1A mutations in episodic ataxia type 2 (EA2), to search for mutations in sporadic cases, and to delineate better the clinical spectrum. BACKGROUND: EA2 is an autosomal dominant disorder characterized by recurrent acetazolamide-responsive attacks of cerebellar ataxia. The mutated gene, CACNA1A, located on chromosome 19, encodes the alpha1A subunit of a voltage-dependent calcium channel. So far, only three CACNA1A mutations have been identified-in two EA2 families and in one sporadic case. These three mutations disrupted the reading frame and led to truncated proteins. Interestingly, distinct types of CACNA1A mutations have been identified in familial hemiplegic migraine (missense mutations) and spinocerebellar ataxia type 6 (SCA-6) progressive cerebellar ataxia (expanded CAG repeats). However, except for SCA-6, these genotype-phenotype correlations relied on the analysis of very few families. METHODS: To characterize CACNA1A mutations, eight familial and seven sporadic EA2 patients were selected. All 47 exons of CACNA1A were screened by a combination of single-strand conformer polymorphism and sequencing analysis. In addition, the length of the CAG repeat has been determined in all patients. RESULTS: Seven new mutations were detected in four multiple case families and three sporadic cases. Six of them lead most likely to truncated or aberrant proteins. CAG repeat sizes were in the normal range. CONCLUSION: These data clearly establish the specificity of EA2 mutations compared with SCA-6 and familial hemiplegic migraine. Detailed clinical analysis of the mutation carriers showed the highly variable penetrance and expression of this disorder: Several of the carriers did not show any clinical symptom; others displayed atypical or permanent neurologic symptoms (such as recurrent, transient diplopia or severe, permanent, and isolated cerebellar ataxia). PMID- 10371529 TI - Definitive molecular diagnosis of facioscapulohumeral dystrophy. AB - OBJECTIVE: To establish the usefulness of a molecular diagnostic protocol for the autosomal dominant disease facioscapulohumeral dystrophy (FSHD). BACKGROUND: The genetic defect underlying the majority of cases is a deletion on chromosome 4q35 that is not associated with the coding sequence of any known gene. Molecular diagnosis of FSHD involves the visualization of this deletion as a "small" EcoRI restriction fragment. However, molecular diagnostics are complicated because of the homology of the telomeric regions of chromosomes 4q and 10q; the homologous 10q26 EcoRI fragments are also detected, and can fall into the size range considered to be diagnostic for FSHD. It is therefore important to distinguish the 4q35 and 10q26 EcoRI fragments, taking advantage of the presence of additional restriction sites (BlnI) in the alleles of chromosome 10q origin. METHODS: Paired digests of genomic DNA (EcoRI only and EcoRI/BlnI double digest), followed by pulsed field gel electrophoresis (PFGE), were used to establish the molecular diagnosis of FSHD in 82 unrelated index cases (46 familial, 24 proven sporadic with de novo mutations, and 12 with uncertain family history). RESULTS: In all cases fulfilling FSHD diagnostic criteria, a 4q35 EcoRI allele size of < or = 38 kb was present. The smallest 4q35 EcoRI allele in 205 normal control subjects was 41 kb. EcoRI alleles < or = 38 kb of chromosome 10q26 origin were present in 11.2% of this control group. In problematic cases, it was possible to resolve the diagnostic question. CONCLUSIONS: The combination of double digestion with EcoRI and BlnI followed by PFGE is the most reliable molecular protocol for distinguishing patients with FSHD. PMID- 10371530 TI - Novel missense mutation in the early growth response 2 gene associated with Dejerine-Sottas syndrome phenotype. AB - BACKGROUND: Mutations in the early growth response 2 (EGR2) gene have recently been found in patients with congenital hypomyelinating neuropathy and Charcot Marie-Tooth type 1 (CMT1) disease. OBJECTIVE: To determine the frequency of EGR2 mutations in patients with a diagnosis of CMT1, Dejerine-Sottas syndrome (DSS), or unspecified peripheral neuropathies. METHODS: Fifty patients and 70 normal control subjects were screened. RESULTS: A de novo missense mutation (Arg359Trp) in the alpha-helix of the first zinc-finger domain of the EGR2 transcription factor was identified in a patient diagnosed with a clinical phenotype consistent with DSS. This patient had a motor median nerve conduction velocity of 8 m/s. A sural nerve biopsy showed a severe loss of myelinated and unmyelinated fibers, evidence for demyelination, numerous classic onion bulbs, and focally folded myelin sheaths. DSS is a severe, childhood-onset demyelinating peripheral neuropathy initially thought to be inherited as an autosomal recessive trait. However, several dominant heterozygous mutations in the peripheral myelin protein 22 (PMP22) gene and dominant mutations in the peripheral myelin protein zero (MPZ) gene, both in the heterozygous and homozygous state, have been reported in patients with DSS. CONCLUSIONS: Hereditary peripheral neuropathies represent a spectrum of disorders due to underlying defects in myelin structure or formation. PMID- 10371531 TI - I.v. immunoglobulin reduces circulating proinflammatory cytokines in Guillain Barre syndrome. AB - BACKGROUND: Treatment with human i.v. immunoglobulin (IVIg) modifies the course of Guillain-Barre syndrome (GBS), but its specific mode of action is unknown. Cellular interactions mediated through the release of cytokines play a role in the pathogenesis of GBS and may be regulated by IVIg therapy. OBJECTIVE: To delineate possible immunoregulatory mechanisms of IVIg in patients with GBS. METHODS: Circulating levels of the proinflammatory cytokines, tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta, were assayed in 21 patients with GBS before and serially after IVIg therapy. Comparisons were made with serum concentration of the anti-inflammatory cytokines, soluble TNF-alpha receptor and IL-10. Serial measurements were also performed in 12 untreated patients with relatively mild disease and 7 patients treated by plasma exchange. RESULTS: Circulating levels of TNF-alpha and IL-1beta decreased after treatment with IVIg but remained relatively high in untreated patients and in those treated by plasma exchange. Clinical improvement in patients treated with IVIg was associated with a reduction in unbound TNF-alpha during the acute phase of the illness. Circulating levels of anti-inflammatory cytokines were not affected by IVIg treatment. CONCLUSION: Data presented here suggest a novel mechanism of action of IVIg that involves selective modulation of circulating proinflammatory cytokines. PMID- 10371532 TI - Comparison of Lewy body variant of Alzheimer's disease with pure Alzheimer's disease: Consortium to Establish a Registry for Alzheimer's Disease, Part XIX. AB - OBJECTIVE: To compare the clinical, neuropsychological, and neuropathologic findings in patients with AD alone with those in patients with the Lewy body variant of AD (LBV). BACKGROUND: Prior studies indicate that patients with LBV not only have distinct clinical and neuropsychological differences from those with AD alone, but have a poorer prognosis with shorter survival time. METHODS: The authors evaluated 74 patients with autopsy-confirmed AD alone and 27 patients with LBV, and compared demographic characteristics and clinical, neuropsychological, and neuropathologic findings. RESULTS: The two groups of patients were equivalent with respect to age at time of entry into the study, years of education, and sex. Two or more extrapyramidal clinical manifestations were found in 44% of patients with LBV, compared with 16% of patients with AD alone (p = 0.02). Duration of survival after entry into the study was similar in both groups, with a mean survival of 3.6 (+/-2.1) years for AD alone versus 3.8 (+/-1.9) years for LBV. Of the various neuropsychological tests administered at the last Consortium to Establish a Registry for Alzheimer's Disease evaluation, only delayed recall of a learned word list was significantly different in the two groups, with 32% of patients with LBV versus 15% of patients with AD alone recalling any items (p = 0.04). Neuropathologic findings confirmed those of previous studies and showed that neurofibrillary tangles were significantly less frequent in the neocortex of patients with LBV than in those with AD alone. CONCLUSION: Compared with patients with AD alone, those with LBV had a greater frequency of extrapyramidal manifestations, somewhat better recall on a selected memory task at their final evaluation, and a significantly lower frequency of neocortical neurofibrillary tangles at autopsy. There were no differences between the two groups, however, in survival time from entry into the study. PMID- 10371533 TI - Misrepresentation of horizontal space in left unilateral neglect: role of hemianopia. AB - BACKGROUND: Right-brain-damaged patients with left unilateral neglect are reported to misperceive the horizontal extension of contralesional stimuli as being shorter than that of ipsilesional stimuli. OBJECTIVE: To investigate the functional and anatomic correlates of horizontal space misrepresentation. METHODS: Eight right-brain-damaged patients with contralesional neglect and complete hemianopia (N+H+), nine right-brain-damaged patients with contralesional neglect and no visual field defect (N+H-), and five unilateral brain-damaged patients with contralesional complete hemianopia and no neglect (N-H+) reproduced a horizontal distance (10 cm) in the contralesional and ipsilesional hemispace. RESULTS: N+H+ patients overextended the distance contralesionally and underextended the same distance ipsilesionally. N+H- and N-H+ patients reproduced equivalent distances contralesionally and ipsilesionally. Compared with N+H- patients, N+H+ patients had a greater ipsilesional shift when bisecting horizontal lines; however, these two groups of patients had comparable neglect severity on multiple-item cancellation tasks. In the N+H+ group the area of maximal overlapping of the lesion was in the posterior cerebral lobes. CONCLUSION: Complete contralesional hemianopia after posterior brain damage is an important factor in determining misrepresentation of horizontal space in patients with left unilateral neglect. PMID- 10371534 TI - Procedural learning is impaired in patients with prefrontal lesions. AB - OBJECTIVES: To 1) determine the effect of prefrontal cortex lesions on procedural learning (PL), measured by a serial reaction-time task (SRTT); 2) confirm whether visuomotor PL is lateralized to one hemisphere; and 3) clarify the relation between visuomotor sequence learning and verbal sequence learning, working memory, and executive functions. BACKGROUND: Previous cognitive neuroscience research has implicated the prefrontal cortex in visuomotor PL but there is a lack of studies examining patients with prefrontal cortex lesions. METHODS: We studied 22 patients with strictly unilateral prefrontal cortex lesions (traumatic, ischemic, hemorrhagic, or tumors) and 52 cognitively intact controls matched for age, sex, and educational level. We administered to subjects long (10 item sequence) and short (4-item sequence) versions of the SRTT. With the long version, each hand was evaluated separately. Learning was indicated by the shortening of response times (RT) and decrease in errors across the sequential blocks and, most importantly, the rebound increase in RTs and errors when comparing the last sequence block with the next random block. Frontal lobe functions and verbal sequence learning were also assessed. RESULTS: Patients with unilateral prefrontal cortex lesions show PL impairment that involves both hands, although more errors were observed when the hand contralateral to the lesion was performing. Only those patients whose lesions were >2 cm in diameter were impaired. Neuropsychologic evaluation indicated impaired verbal sequence learning and executive function deficits. Patients with poorer working memory and verbal sequence learning were also more impaired in visuomotor sequence learning. CONCLUSIONS: The prefrontal cortex has a role in PL and is part of the neural circuit that mediates this type of learning. PMID- 10371535 TI - Brain regions associated with episodic retrieval in normal aging and Alzheimer's disease. AB - OBJECTIVE: To examine patterns of brain activation during verbal episodic retrieval in normal elderly subjects and patients in an early phase of AD. BACKGROUND: It is established that 1) a profound episodic memory impairment is a cardinal symptom of AD; and 2) some of the earliest brain changes in this disease occur in regions critical to episodic memory, such as the hippocampus and neighboring regions. Yet, it remains largely unknown whether the episodic memory deficit seen in AD is paralleled by concomitant alterations in brain activity during actual task performance in these or other brain areas. METHODS: Using PET, blood flow was assessed in normal elderly subjects and patients with early AD during two retrieval conditions involving completion of word stems: baseline and cued recall. RESULTS: The patients with AD showed a marked performance deficit in cued recall, although the two groups were indistinguishable in the baseline task condition. Both groups showed bilateral activity in orbital and dorsolateral prefrontal cortex, left precuneus, and right cerebellum, as well as decreased activity in distinct left temporal regions during cued recall. The normal elderly alone activated the left parietal cortex and the left hippocampal formation during episodic retrieval. By contrast, AD-related increases in activity during cued recall were observed in the left orbital prefrontal cortex and left cerebellum. CONCLUSIONS: The similar patterns of activations in the two groups suggest that a large distributed network involved in episodic memory retrieval functions relatively normally in early AD. Those retrieval activations seen in the normal elderly, as opposed to the patients, may reflect AD-related failures in semantic processing and successful recollection of the target information, respectively. Finally, the AD-related increases in activity were interpreted in terms of compensatory reactions to the difficulties in performing the episodic memory task. PMID- 10371536 TI - Entorhinal cortex in temporal lobe epilepsy: a quantitative MRI study. AB - BACKGROUND: The entorhinal cortex (EC) is a distinct anatomic and functional region of the anterior parahippocampal gyrus, which plays a role in seizure generation and propagation in temporal lobe epilepsy (TLE). In tissue resected from TLE patients, cell loss in the EC has been described. OBJECTIVES: To develop a standardized protocol for identifying the anatomic boundaries of the EC using high-resolution MRI and to examine morphologic changes of the EC in TLE. METHODS: We performed T1-weighted MRIs in 20 patients (7 males) with TLE (mean age 34 years) and 18 normal controls (mean age 26 years). Eleven patients had a left and 9 a right epileptic focus as defined by history, video-EEG, and surgical outcome. The volumes of the EC, the hippocampus, and the amygdala were measured using a standardized MRI protocol. Analysis of variance (ANOVA) was used to examine the effect of seizure focus lateralization and hemisphere on these volumes. An asymmetry ratio [A (%) = 100 x (R-L)/(R+L)/2] was also compared between groups using ANOVA. RESULTS: In normal controls the volume of the right EC was 1,247 +/- 127 mm3 (mean +/- standard deviation), and that of the left EC was 1,215 +/- 135 mm3 (p > 0.05). We found a bilateral reduction in the volume of the EC in TLE patients compared with controls (p < 0.05). Examination of the asymmetry ratios showed that the reduction in volume of the EC was greater ipsilateral to the epileptic focus (p < 0.05). The volumes of the hippocampus and the amygdala were smaller on the side of the focus in TLE patients compared with controls (p < 0.05). CONCLUSIONS: With a standardized protocol for the quantitative assessment of the EC, patients with unilateral TLE show bilateral reduction in the volume of the EC. However, this reduction is more severe ipsilateral to the epileptic focus. PMID- 10371537 TI - Familial aggregation of Parkinson's disease: a population-based case-control study in Europe. EUROPARKINSON Study Group. AB - OBJECTIVE: To investigate the familial aggregation of PD in a large collaborative population-based case-control study. BACKGROUND: Most previous case-control studies of the familial aggregation of PD have been hospital- or clinic-based. METHODS: We included 219 prevalent cases ascertained in three European populations (centers), using a two-phase design consisting of screening and examination by a neurologist. Each case was matched by age, sex, and center to three controls drawn from the same populations (n = 657). Presence of PD among first-degree relatives (parents and siblings) was determined using the family history approach for 175 cases and 481 controls. RESULTS: Overall, a positive family history (at least one parent or sibling affected by PD) was reported in 10.3% of patients and 3.5% of controls (odds ratio [OR] = 3.2; 95% confidence interval [CI] = 1.6 to 6.6). A similar association was observed when analyses were restricted to nondemented patients and controls (OR = 3.9; 95% CI = 1.7 to 8.7) or to newly diagnosed patients (OR = 3.3; 95% CI = 0.9 to 11.9). We found a significant trend of increasing risk with increasing number of affected relatives (p = 0.003). Analyses stratified by age showed a stronger association for younger PD patients (OR = 7.6; 95% CI = 1.5 to 38.9) than for older patients (OR = 2.5; 95% CI = 1.1 to 5.7). CONCLUSIONS: In this large sample of prevalent PD patients and population-matched controls, PD significantly aggregates in families, with the strength of the association being age-dependent. Therefore, familial factors, which can be genetic, environmental, or both, play a role in PD. PMID- 10371539 TI - The cerebellar seizures of Hughlings Jackson. PMID- 10371538 TI - A double-blind, randomized trial of topiramate in Lennox-Gastaut syndrome. Topiramate YL Study Group. AB - OBJECTIVE: To evaluate the efficacy and safety of topiramate as adjunctive therapy for Lennox-Gastaut syndrome in a multicenter, double-blind, placebo controlled trial. BACKGROUND: Conventional antiepileptic drugs are frequently ineffective against multiple-seizure types of Lennox-Gastaut syndrome. METHODS: Ninety-eight patients >1 year to <30 years of age, with slow spike-and-wave patterns on EEG, seizure types including drop attacks, and either a history of or active atypical absence seizures, were assigned to an 11-week, double-blind treatment phase with either topiramate or placebo. Topiramate was titrated to target doses of approximately 6 mg/kg/d. RESULTS: For drop attacks, the most severe seizures associated with this syndrome, the median percentage reduction from baseline in average monthly seizure rate was 14.8% for the topiramate group and -5.1% (an increase) for the placebo group (p = 0.041). Topiramate-treated patients demonstrated greater improvement in seizure severity than did placebo treated patients based on parental global evaluations (p = 0.037). The percentage of patients with a > or = 50% reduction from baseline in major seizures (drop attacks and tonic-clonic seizures) was greater in the topiramate group (15/46 or 33%) than in the control group (4/50 or 8%; p = 0.002). The most common adverse events in both groups were CNS related; there were no discontinuations from topiramate therapy due to adverse events. CONCLUSIONS: Topiramate adjunctive therapy was effective in reducing the number of drop attacks and major motor seizures and in improving seizure severity as determined by parental global evaluation. PMID- 10371540 TI - Neuro@neurolist.mcg.edu: an e-mail discussion list for neurologists. AB - INeuro@neurolist.mcg. edu: An e-mail discussion list for neurologists Neuro@neurolist.mcg.edu (Neurolist) is a discussion list for neurologists. This Internet tool, established in 1994, allows the worldwide community of neurologists to communicate quickly and efficiently using e-mail, web browsers, and newsreaders. Neurolist currently has 584 members in 51 countries. Between June 1997 and October 1998, 6179 messages were transmitted by Neurolist. Most of the list's posts (70%) concerned patient care and neurologic diseases. Users may subscribe at the web site: . The service is free. PMID- 10371541 TI - Ibuprofen treatment versus gradual introduction of interferon beta-1b in patients with MS. AB - Flu-like symptoms and injection site reactions are adverse effects of treatment with interferon beta-1b in patients with MS. We compared gradual dose escalation, ibuprofen treatment, or their combination in an open-label study. The combination reduced the incidence of flu-like symptoms to rates comparable with the placebo group in the pivotal trial but increased the frequency of injection site reactions, albeit modestly and transiently. PMID- 10371542 TI - Relevance of interleukin 1 receptor antagonist intron 2 polymorphism in Italian MS patients. AB - The A1/A1 genotype of the anti-inflammatory cytokine interleukin 1 receptor antagonist (IL-1Ra) polymorphism was more frequent in 339 Italian MS patients than in healthy controls (HCs) (odds ratio = 1.83). A more aggressive disease course was also associated with A1+ genotypes and might reflect the reduced ability of mononuclear cell cultures of A1+ HCs to produce IL-1Ra. We conclude that an IL-1Ra gene polymorphism is associated with occurrence of disease and clinical course variability in Italian MS patients. PMID- 10371543 TI - Molecular genetic analysis of the APEX nuclease gene in amyotrophic lateral sclerosis. AB - We analyzed genomic DNA from ALS patients for mutations in the apurinic/apyrimidinic endonuclease (APEX nuclease) gene. We identified three rare polymorphisms in the untranslated region of the gene and one common two-allele polymorphism (D148E). The allelic frequency D148E was significantly different in sporadic ALS patients compared with controls. A conserved amino acid change and a 4-base pair deletion were also identified in sporadic ALS patients. These data suggest that APEX nuclease may contribute to the etiology of ALS. PMID- 10371544 TI - Recurrent multifocal demyelinating neuropathy with febrile illness and IgG subset deficiency. AB - We describe a unique syndrome of recurrent multifocal demyelinating motor greater than sensory deficits in cranial and peripheral nerve distributions with rapid, spontaneous improvement. Three patients presented with episodes over a period of 7 to 24 years, largely accompanied by febrile illness. Variably decreased IgG1 and IgG3 subclass levels were found. We postulate an immune-mediated process based upon the clinical presentation and presence of decreased IgG subclass levels. PMID- 10371545 TI - Maternally inherited hearing loss in a large kindred with a novel T7511C mutation in the mitochondrial DNA tRNA(Ser(UCN)) gene. AB - Thirty-six of 43 maternally related members of a large African American family experienced hearing loss. A muscle biopsy specimen from the proband showed cytochrome c oxidase (COX)-deficient fibers but no ragged-red fibers; biochemical analysis showed marked reduction of COX activity. A novel T7511C point mutation in the tRNA(Ser(UCN)) gene was present in almost homoplasmic levels (>95%) in the blood of 18 of 20 family members, and was also found in lower abundance in the other two. Single-fiber PCR showed that the mutational load was greater in COX deficient muscle fibers. The tRNA(ser(UCN)) gene may be a "hot spot" for mutations associated with maternally transmitted hearing loss. PMID- 10371546 TI - Falling asleep at the wheel: motor vehicle mishaps in persons taking pramipexole and ropinirole. AB - The authors report a new side effect of the dopamine agonists pramipexole and ropinirole: sudden irresistible attacks of sleep. Eight PD patients taking pramipexole and one taking ropinirole fell asleep while driving, causing accidents. Five experienced no warning before falling asleep. The attacks ceased when the drugs were stopped. Neurologists who prescribe these drugs and patients who take them should be aware of this possible side effect. PMID- 10371547 TI - Bilateral perisylvian polymicrogyria in three generations. AB - A family is described in which bilateral perisylvian polymicrogyria was present in 6 members of 3 consecutive generations. Typical anatomic and clinical findings of the syndrome, with a mild phenotype, were present in the 5 affected women from all 3 generations. More severe impairment was observed in the only affected male individual, a boy, in the third generation. Analysis of the pedigree and severity of the phenotype in the affected boy are consistent with transmission of an X linked dominant trait, although other patterns of inheritance cannot be ruled out with certainty. PMID- 10371548 TI - Diagnostic Notch3 sequence analysis in CADASIL: three new mutations in Dutch patients. Dutch CADASIL Research Group. AB - To confirm the clinical diagnosis in individual Dutch patients with cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), we performed direct sequence analysis of the abnormal gene, Notch3, in patients from 11 families without prior linkage analysis to chromosome 19. Eleven missense mutations involving the loss or gain of a cysteine residue were found, of which 3 are new. Exon 4 is a mutation hotspot (9 of 11 families). Notch3 sequence analysis of CADASIL patients in a diagnostic laboratory is a feasible procedure to confirm the clinical diagnosis in individual patients. PMID- 10371549 TI - Theophylline increases "on" time in advanced parkinsonian patients. PMID- 10371550 TI - Topiramate relieves refractory intercostal neuralgia. PMID- 10371551 TI - Progesterone therapy in women with epilepsy: a 3-year follow-up. PMID- 10371552 TI - Elimination of oxcarbazepine-induced oculogyric crisis following vagus nerve stimulation. PMID- 10371553 TI - A randomized, double-blind, placebo-controlled trial of deprenyl and thiotic acid in HIV-associated cognitive impairment. PMID- 10371554 TI - A randomized, double-blind, placebo-controlled trial of deprenyl and thiotic acid in HIV-associated cognitive impairment. PMID- 10371555 TI - Micrographia after thalamo-mesencephalic infarction. PMID- 10371556 TI - Micrographia after thalamo-mesencephalic infarction. PMID- 10371557 TI - Mechanical ventilation for ischemic stroke and intracerebral hemorrhage. PMID- 10371558 TI - Major histocompatibility complex class II alleles and the course and outcome of MS. PMID- 10371559 TI - Sleep-related violence, injury, and REM sleep behavior disorder in PD. PMID- 10371560 TI - Information versus choice in infertility treatment. PMID- 10371561 TI - Breast screening in women aged 40-49 years: what next? PMID- 10371562 TI - A "breathalyser" for lung cancer? PMID- 10371563 TI - Effect of growth-hormone therapy on early atherosclerotic changes in GH-deficient adults. PMID- 10371564 TI - Screening methods for congenital toxoplasma and risk of disease. PMID- 10371565 TI - Sexual dysfunction after surgery for rectal cancer. PMID- 10371566 TI - Hospitalism in the USA. PMID- 10371567 TI - 14 years of follow-up from the Edinburgh randomised trial of breast-cancer screening. AB - BACKGROUND: The Edinburgh randomised trial of breast-cancer screening recruited women aged 45-64 years from 1978 to 1981 (cohort 1), and those aged 45-49 years during 1982-85 (cohorts 2 and 3). Results based on 14 years of follow-up and 270,000 woman-years of observation are reported. METHODS: Breast-cancer mortality rates in the intervention group (28,628 women offered screening) were compared with those in the control group (26,026) with adjustment for socioeconomic status (SES) of general medical practices. Rate ratios were derived by means of logistic regression for the total trial population and for women first offered screening while younger than 50 years. Analyses were by intention to treat. FINDINGS: Initial unadjusted results showed a difference of just 13% in breast-cancer mortality rates between the intervention and control groups (156 deaths [5.18 per 10,000] vs 167 [6.04 per 10,000]; rate ratio 0.87 [95% CI 0.70-1.06]), but the results were influenced by differences in SES by trial group. After adjustment for SES, the rate ratio was 0.79 (95% CI 0.60-1.02). When deaths after diagnosis more than 3 years after the end of the study were censored the rate ratio became 0.71 (0.53-0.95). There was no evidence of heterogeneity by age at entry and no evidence that younger entrants had smaller or delayed benefit (rate ratio 0.70 [0.41-1.20]). No breast-cancer mortality benefit was observed for women whose breast cancers were diagnosed when they were younger than 50 years. Other-cause mortality rates did not differ by trial group when adjusted for SES. INTERPRETATION: Our findings confirm results from randomised trials in Sweden and the USA that screening for breast cancer lowers breast-cancer mortality. Similar results are reported by the UK geographical comparison, UK Trial of Early Detection of Breast Cancer. The results for younger women suggest benefit from introduction of screening before 50 years of age. PMID- 10371568 TI - 16-year mortality from breast cancer in the UK Trial of Early Detection of Breast Cancer. AB - BACKGROUND: The UK Trial of Early Detection of Breast Cancer (TEDBC) is a non randomised study, which was set up in 1979 to investigate the effect of screening and education about breast self-examination on breast-cancer mortality. We report mortality results after 16 years of follow-up, including results by age at trial entry. METHODS: Eight centres (two screening, two breast self-examination, and four comparison) in England and Scotland recruited women aged 45-64 years into the initial cohort, with women reaching age 45 years during the 7-year study period included in later cohorts. The observed number of deaths from breast cancer in each centre was compared with the expected number, which was calculated by Poisson regression model; expected numbers were adjusted for pretrial breast cancer mortality. Results were analysed by 5-year age-groups, and for women aged 45-46 years and 47-49 years at entry. FINDINGS: Breast-cancer mortality was 27% lower (rate ratio 0.73 [95% CI 0.63-0.84]), adjusted for pretrial rates, in cohort 1 in the two screening centres combined than in the comparison centres. No reduction in mortality in the two breast self-examination centres combined was seen (rate ratio 0.99 [0.87-1.12]). The mortality reduction in the screening centres did not differ significantly across age-groups; a 35% reduction was seen in women in all cohorts aged 45-46 years at entry (rate ratio 0.65 [0.50-0.86]). Results were similar when deaths were restricted to those in patients diagnosed within 10 years of trial entry. INTERPRETATION: The results from TEDBC support those from randomised trials in Edinburgh and elsewhere, and show that a reduction in breast-cancer mortality resulting from screening can be achieved in the UK. There was no evidence of less benefit in women aged 45-46 years at the start of screening; the effect of screening in this age-group begins to emerge after 3-4 years. PMID- 10371569 TI - Effect of simultaneous pancreas-kidney transplantation on mortality of patients with type-1 diabetes mellitus and end-stage renal failure. AB - BACKGROUND: Long-term prognosis of patients with type-1 diabetes mellitus and end stage renal failure appears to be better after kidney transplantation compared with dialysis. Controversy exists about the additional benefit of a simultaneously transplanted pancreatic graft. We studied the effect on mortality of simultaneous pancreas-kidney transplantation compared with kidney transplantation alone from regional differences in transplantation protocols. METHODS: All 415 patients with type-1 diabetes (aged 18-52 years) who started renal-replacement therapy in the Netherlands between 1985 and 1996 were included in the analysis. Patients were allocated to a centre based on their place of residence at onset of renal failure. In the Leiden area, the primary intention to treat was with a simultaneous pancreas-kidney transplantation, whereas in the non Leiden area, kidney transplantation alone was the predominant type of treatment. All patients were followed up to July, 1997. Analyses, mortality, and graft failure were by Cox proportional-hazard model adjusted for age and sex. FINDINGS: Simultaneous pancreas-kidney transplantation was done in 41 (73%) of 56 transplanted patients in the Leiden area compared with 59 (37%) of 158 transplanted patients in the non-Leiden area (p<0.001). The hazard ratio for mortality after the start of renal-replacement therapy was 0.53 (95% CI, 0.36 0.77, p<0.001) in the Leiden area compared with the non-Leiden area. When just the transplanted patients were analysed the mortality ratio was 0.4 (95% CI 0.20 0.77, p=0.008) and was independent of duration of dialysis and early transplant related deaths. Equal survival was found for patients on dialysis only. INTERPRETATION: These data support the hypothesis that simultaneous pancreas kidney transplantation prolongs survival in patients with diabetes and end-stage renal failure. PMID- 10371570 TI - Mortality in severely malnourished children with diarrhoea and use of a standardised management protocol. AB - BACKGROUND: Severely malnourished children have high mortality rates. Death commonly occurs during the first 48 h after hospital admission, and has been attributed to faulty case-management. We developed a standardised protocol for acute-phase treatment of children with severe malnutrition and diarrhoea, with the aim of reducing mortality. METHODS: We compared severely malnourished children with diarrhoea aged 0-5 years managed by non-protocol conventional treatment, and those treated by our standardised protocol that included slow rehydration with an emphasis on oral rehydration. The standardised-protocol group included children admitted to the ICDDR,B Hospital, Dhaka between Jan 1, 1997, and June 30, 1997, while those admitted between Jan 1, 1996, and June 30, 1996, before the protocol was implemented, were the non-protocol group. FINDINGS: Characteristics on admission of children on standardised protocol (n=334) and non protocol children (n=293) were similar except that more children on standardised protocol had oedema, acidosis, and Vibrio cholerae isolated from stools. 199 (59.9%) of children on standardised protocol were successfully rehydrated with oral rehydration solution, compared with 85 (29%) in the non-protocol group (p<0.0001). Use of expensive antibiotics was less frequent in children on standardised protocol than in the other group (p<0.0001). Children on standardised protocol had fewer episodes of hypoglycaemia than non-protocol children (15 vs 30, p=0.005). 49 (17%) of children on non-protocol treatment died, compared with 30 (9%) children on standardised protocol (odds ratio for mortality, 0.49, 95% CI 0.3-0.8, p=0.003). INTERPRETATION: Compared with non protocol management, our standardised protocol resulted in fewer episodes of hypoglycaemia, less need for intravenous fluids, and a 47% reduction in mortality. This standardised protocol should be considered in all children with diarrhoea and severe malnutrition. PMID- 10371571 TI - Comparison of subcutaneous and intravenous interleukin-2 in asymptomatic HIV-1 infection: a randomised controlled trial. ANRS 048 study group. AB - BACKGROUND: Intermittent interleukin-2 therapy for HIV-1 by continuous intravenous infusion leads to sustained increase of CD4 T cells. This method of administration is, however, inconvenient and has limiting toxic effects. We did a randomised study to compare safety and efficacy of antiviral treatment alone or combined with various interleukin-2 regimens in HIV-1-infected patients. METHODS: 94 symptom-free patients, naive to antiretroviral treatment, with CD4-T-cell counts of 250-550 cells/microL at baseline were randomly assigned zidovudine and didanosine alone (n=26) or combined with interleukin-2 administered intravenously (12 million IU/day, n=22) or subcutaneously (3 million IU/m2 twice daily, n=24) for 5 days, or were given polyethylene-glycol-modified (PEG) interleukin-2 (2 million IU/m2 intravenous bolus, n=22) administered every 2 months from week 2 to week 50 (seven cycles). Safety and immunological and virological results were monitored until week 56. FINDINGS: CD4-T-cell count increased to higher than baseline by a mean of 564 cells/microL (subcutaneous group), 676 cells/microL (intravenous group), 105 cells/microL (PEG group), and 55 cells/microL (antiretroviral-therapy group, p=0.0001). 68% and 77% of patients in the subcutaneous and intravenous groups, respectively, achieved an 80% increase of CD4 T cells (p<0.001). In these two groups, 50% of patients restored a CD4/CD8-T cell ratio of more than 1. The groups did not differ significantly for changes in plasma HIV-1 RNA loads throughout the study. The duration of common side-effects of interleukin-2 was shorter in the subcutaneous group, which enabled outpatient treatment. Naive and memory CD4 T cells, CD28 expression on CD4 and CD8 T cells, and restoration of in-vitro proliferative response to mitogens and recall antigens increased in the intravenous and subcutaneous groups. INTERPRETATION: Subcutaneous interleukin-2 is a convenient regimen that, as well as intravenous therapy, improves immunological function in HIV-1-infected patients receiving two nucleosides. Larger studies are needed to show whether immunological improvements translate into clinical benefit. PMID- 10371573 TI - A large bruise. PMID- 10371572 TI - Volatile organic compounds in breath as markers of lung cancer: a cross-sectional study. AB - BACKGROUND: Many volatile organic compounds (VOCs), principally alkanes and benzene derivatives, have been identified in breath from patients with lung cancer. We investigated whether a combination of VOCs could identify such patients. METHODS: We collected breath samples from 108 patients with an abnormal chest radiograph who were scheduled for bronchoscopy. The samples were collected with a portable apparatus, then assayed by gas chromatography and mass spectroscopy. The alveolar gradient of each breath VOC, the difference between the amount in breath and in air, was calculated. Forward stepwise discriminant analysis was used to identify VOCs that discriminated between patients with and without lung cancer. FINDINGS: Lung cancer was confirmed histologically in 60 patients. A combination of 22 breath VOCs, predominantly alkanes, alkane derivatives, and benzene derivatives, discriminated between patients with and without lung cancer, regardless of stage (all p<0.0003). For stage 1 lung cancer, the 22 VOCs had 100% sensitivity and 81.3% specificity. Cross-validation of the combination correctly predicted the diagnosis in 71.7% patients with lung cancer and 66.7% of those without lung cancer. INTERPRETATION: In patients with an abnormal chest radiograph, a combination of 22 VOCs in breath samples distinguished between patients with and without lung cancer. Prospective studies are needed to confirm the usefulness of breath VOCs for detecting lung cancer in the general population. PMID- 10371574 TI - Rapid ambulance protocol for acute stroke. PMID- 10371575 TI - Brill-Zinsser disease in France. PMID- 10371576 TI - Plasma homocysteine and all-cause mortality in diabetes. PMID- 10371577 TI - The sentinel node in breast cancer: acceptable false-negative rate. PMID- 10371578 TI - Ultrasound-imaging of lateral thoracic arteries to detect breast cancer. PMID- 10371579 TI - High bioavailability of reduced iron added to UK flour. PMID- 10371580 TI - Attenuation of heart-rate variability in postmenopausal women on progestin containing hormone replacement therapy. PMID- 10371581 TI - Neonatal seizures after use of pyridoxine in pregnancy. PMID- 10371582 TI - Twinning and maternal risk of ovarian cancer. PMID- 10371583 TI - House-dust mite and cat allergens in the Antarctic. PMID- 10371584 TI - A Ca++ activated serine protease (LOPAP) could be responsible for the haemorrhagic syndrome caused by the caterpillar Lonomia obliqua. L obliqua Prothrombin Activator Protease. PMID- 10371585 TI - Taking the bite out of snake venoms. PMID- 10371586 TI - Britain on the brink of new heroin epidemic. PMID- 10371587 TI - Human T-lymphotropic virus type I infection. AB - Human T-cell lymphotropic virus type I (HTLV-I) is the first human retrovirus to be associated with malignant disease--namely, adult T-cell leukaemia/lymphoma. HTLV-I has also been associated with several non-malignant conditions, notably the chronic neurodegenerative disorder, HTLV-I associated myelopathy (also known as tropical spastic paraparesis), infective dermatitis of children and uveitis. More recent evidence points to disease associations not previously linked to HTLV I. Thus, the disease spectrum of HTLV-I is not fully known. HTLV-I has a worldwide distribution with major endemic foci in the Caribbean and southern Japan. The public health importance is confirmed by the major routes of transmission, which are mother-to-child, blood transfusion, and sexual activity. Unfortunately, no vaccine is available yet and there is no proven treatment for advanced HTLV-I disease. PMID- 10371588 TI - Neuropathic pain: aetiology, symptoms, mechanisms, and management. AB - We highlight current theories about peripheral neuropathic pain and show that progress in management is contingent on targeting treatment not at the aetiological factors or the symptoms but at the mechanisms that operate to produce the symptoms. This approach will require substantial progress in our understanding of the pathophysiology of neuropathic pain, the development of accurate diagnostic tools to discover what mechanisms contribute to the pain syndrome in an individual, and effective treatments aimed specifically at the mechanisms. PMID- 10371589 TI - Averting a malaria disaster. PMID- 10371590 TI - Glucose-insulin-potassium in acute myocardial infarction. PMID- 10371591 TI - Glucose-insulin-potassium in acute myocardial infarction. PMID- 10371592 TI - Age of entry to nursery and allergies in later life. PMID- 10371593 TI - Chronic stress in elderly carers of dementia patients and influenza vaccine. PMID- 10371594 TI - Churg-Strauss syndrome. PMID- 10371595 TI - Water drinking and the heart. PMID- 10371596 TI - Warfarin dose requirement and CYP2C9 polymorphisms. PMID- 10371597 TI - Management of cervical intraepithelial neoplasia. PMID- 10371598 TI - Familial papillary thyroid microcarcinoma. PMID- 10371599 TI - Definition and measurement of adherence to antiretroviral drugs in HIV-1-infected patients. PMID- 10371600 TI - Lipid-lowering drugs and homocysteine. PMID- 10371601 TI - Early diagnosis in Duchenne muscular dystrophy. PMID- 10371602 TI - Surgical treatment for incidentally discovered intracranial aneurysms. PMID- 10371603 TI - Beta2-adrenoceptor gene polymorphism and obesity. PMID- 10371604 TI - Common pitfalls in case management. PMID- 10371605 TI - Health care in Iraq. PMID- 10371606 TI - Health-care camps for the poor provide mass sterilisation quota. PMID- 10371607 TI - Use of laboratory animals. PMID- 10371608 TI - Episiotomy: a form of genital mutilation. PMID- 10371609 TI - COX-2 inhibitors. PMID- 10371610 TI - A much misunderstood caduceus and the case for an aesculapion. PMID- 10371612 TI - The Nobel chronicles. 1957: Daniel Bovet (1907-92). PMID- 10371614 TI - Congress of Neurological Surgeons 1998 Presidential Address PMID- 10371615 TI - The natural history of cavernous malformations: a prospective study of 68 patients AB - OBJECTIVE: Cavernous malformations are angiographically occult cerebrovascular malformations found in approximately 0.5% of the population. To help further understand the natural history of these lesions, we prospectively followed 68 patients harboring cavernous malformations. METHODS: The 68 patients in this study were all diagnosed radiographically (67 patients) or surgically (1 patient) and were entered into a patient database. Age, sex, clinical symptoms, seizure frequency, focal neurological deficits, and presence or absence of extralesional hemorrhage were all recorded at presentation. Patients were then followed prospectively to determine the rate of hemorrhage and new-onset seizures. RESULTS: The mean follow-up per patient was 5.2 years, and the total follow-up was 352.9 patient-years. There was an average of 3.4 lesions per patient. Thirteen of the patients (19%) had familial cavernous malformations. Patients with familial disease were more likely to have multiple lesions than patients with sporadic disease (85% versus 25%, respectively [P = 0.001]). Initial presentation included headache (65%), seizures (49%), and focal neurological deficit (46%). Eleven symptomatic, radiologically proven, extralesional hemorrhages occurred during the 352.9 patient-years of follow-up for an overall hemorrhage rate of 3.1% per patient-year. Female patients had a significantly higher prospective hemorrhage rate (4.2% per patient-year versus 0.9% per patient year [P = 0.04]). A history of hemorrhage at presentation was not a risk factor for subsequent hemorrhage during follow-up. The rate of new-onset seizures was 2.4% per patient-year. CONCLUSION: The clinical presentation and prospective hemorrhage rate reported here agree well with findings of other prospective studies. This information, combined with our new-onset seizure rate, should aid clinicians caring for patients with cavernous malformations. PMID- 10371616 TI - Correlation of magnetic resonance characteristics and histopathological type of angiographically occult vascular malformations AB - OBJECTIVE: Histological and radiological classification of vascular malformations has previously been attempted in an effort to understand their nature and predict their biological behavior. There exists a subgroup of vascular malformations that are angiographically occult and share a common magnetic resonance imaging (MRI) appearance but may differ in their behavior. We sought to determine any correlation between MRI features and final histopathological diagnosis. METHODS: We reviewed our series of 72 patients with angiographically occult vascular malformations operated on at Stanford University Medical Center between 1988 and 1993. Radiographic magnetic resonance images and histopathological specimens were retrospectively evaluated for various diagnostic features. RESULTS: Our data indicate that lesions exhibiting a ring of hemosiderin are associated with the presence of a cavernous malformation (CM) component (86% of CMs versus 33% of non CM lesions). A lesion associated with edema, mass effect, or a single prominent blood product on MRI correlates with the presence of an arteriovenous malformation (AVM) component. Sixty-three percent of AVMs and 80% of lesions with partial AVM components showed edema, compared with 8% of CMs and 0% of venous malformations. Sixty percent of AVMs and 63% of lesions with partial AVM components showed a single prominent blood product, compared with 8% of CMs and 0% of venous malformations. Finally, 60% of AVMs exhibited mass effect, compared with 20% of CMs. Additionally, an expansile hemorrhage is suggestive of an AVM. CONCLUSION: This study is the first to demonstrate that a particular MRI appearance of an angiographically occult vascular malformation is suggestive of an AVM component. This may have important implications with regard to the behavior of the lesion and planning of future treatment. PMID- 10371617 TI - Boron neutron capture therapy for glioblastoma multiforme: interim results from the phase I/II dose-escalation studies AB - OBJECTIVE: The primary objective of these Phase I/II dose-escalation studies is to evaluate the safety of boronophenylalanine (BPA)-fructose-mediated boron neutron capture therapy (BNCT) for patients with glioblastoma multiforme (GBM). A secondary purpose is to assess the palliation of GBM by BNCT, if possible. METHODS: Thirty-eight patients with GBM have been treated. Subtotal or gross total resection of GBM was performed for 38 patients (median age, 56 yr) before BNCT. BPA-fructose (250 or 290 mg BPA/kg body weight) was infused intravenously, in 2 hours, approximately 3 to 5 weeks after surgery. Neutron irradiation was begun between 34 and 82 minutes after the end of the BPA infusion and lasted 38 to 65 minutes. RESULTS: Toxicity related to BPA-fructose was not observed. The maximal radiation dose to normal brain varied from 8.9 to 14.8 Gy-Eq. The volume weighted average radiation dose to normal brain tissues ranged from 1.9 to 6.0 Gy Eq. No BNCT-related Grade 3 or 4 toxicity was observed, although milder toxicities were seen. Twenty-five of 37 assessable patients are dead, all as a result of progressive GBM. No radiation-induced damage to normal brain tissue was observed in postmortem examinations of seven brains. The minimal tumor volume doses ranged from 18 to 55 Gy-Eq. The median time to tumor progression and the median survival time from diagnosis (from Kaplan-Meier curves) were 31.6 weeks and 13.0 months, respectively. CONCLUSION: The BNCT procedure used has been safe for all patients treated to date. Our limited clinical evaluation suggests that the palliation offered by a single session of BNCT is comparable to that provided by fractionated photon therapy. Additional studies with further escalation of radiation doses are in progress. PMID- 10371618 TI - Tumor size predicts control of benign meningiomas treated with radiotherapy AB - OBJECTIVE: The goal of this study was to evaluate the effect of postoperative residual tumor size on the outcomes for patients with incompletely resected benign meningiomas who underwent radiotherapy (RT). METHODS: Fifty-four patients with incompletely resected benign meningiomas received postoperative RT between 1984 and 1995. Surgery consisted of partial resection for 43 patients (80%) and biopsy for 11 (20%). All patients underwent postoperative imaging (using computed tomography and/or magnetic resonance imaging), and the median residual tumor size was 3.5 cm (in greatest dimension). Thirty-eight tumors (69%) were <5 cm, and 17 (31%) were > or =5 cm. The median RT dose was 54 Gy (range, 45-60 Gy), delivered in daily fractions of 1.8 to 2 Gy. The median follow-up period was 55 months. RESULTS: The 5-year actuarial progression-free survival (PFS) rate for the entire group was 76%. The only significant predictor of PFS rates was residual tumor size. Large residual tumors (> or =5 cm) exhibited a worse 5-year PFS rate than did small tumors (<5 cm) (40 versus 93%, P < 0.0001). When analyzed as a continuous variable, residual tumor size remained a significant prognostic factor. Age, sex, tumor histological features, tumor location, timing of treatment (immediate versus delayed), extent of resection, and RT dose (<54 Gy versus > or =54 Gy) did not reach prognostic significance. The difference in PFS rates for small and large residual tumors translated into a significant difference in 5-year cause-specific survival rates (65 versus 97%, P = 0.01). CONCLUSION: For incompletely resected benign meningiomas treated with RT, residual tumor size is the most significant predictor of tumor control. Small residual tumors are well controlled with conventional RT doses and techniques. In contrast, more aggressive therapies should be considered for large tumors. PMID- 10371619 TI - Ocular microtremor in brain stem death AB - OBJECTIVE: This study was undertaken to establish whether measurement of ocular microtremor (OMT) activity could be used as a method to establish brain stem death. Presently, the diagnosis of brain stem death can be made using clinical criteria alone. OMT is a high-frequency, low-amplitude physiological tremor of the eye caused by impulses emanating from the brain stem. There have been a number of reports indicating that the recording of OMT may be useful in the assessment of comatose states and in establishing brain stem viability or death. METHODS: We obtained the OMT recordings of 32 patients suspected of having brain stem death using the piezoelectric strain gauge technique. This method involves mounting the piezoelectric probe in a headset and lowering the rubber-tipped end piece onto the anesthetized scleral surface of the subject. The signal produced is recorded on audiomagnetic tape and later played back and analyzed on an electrocardiographic tape analyzer. RESULTS: In 28 patients, initial clinical assessment confirmed the diagnosis of brain stem death and no OMT activity was recorded from these subjects. In three patients in whom initial clinical assessment demonstrated brain stem function, OMT activity was present; when brain stem death was subsequently diagnosed in these three patients, no OMT activity could be demonstrated. In the remaining patient, two of three OMT recordings demonstrated activity in spite of the absence of clinical evidence of brain stem function. A post mortem revealed bacterial cerebritis in this subject. CONCLUSION: The results suggest that OMT is a sensitive method of detecting brain stem life and that it could play an important role in the assessment of brain stem death. PMID- 10371620 TI - Venous and arterial bypass grafts for difficult tumors, aneurysms, and occlusive vascular lesions: evolution of surgical treatment and improved graft results AB - OBJECTIVE: In the treatment of patients with cranial base tumors, unclippable aneurysms, or medically intractable ischemia, it may be necessary to use high flow bypass grafts. The indications, surgical techniques and complications are discussed. METHODS: During a 10-year period, 99 saphenous vein grafts and 3 radial artery grafts were performed for 101 patients, i.e., 72 with neoplasms, 23 with aneurysms, and 6 with ischemia. Clinical follow-up monitoring of the patients was by direct examination or telephone interview, with a mean follow-up period of 41.2 months (range, 5-147 mo). Radiological follow-up monitoring was by magnetic resonance imaging, magnetic resonance angiography, or three-dimensional computed tomographic angiography, with a mean follow-up period of 32 months (range, 1-120 mo). During the follow-up period, there was one late graft occlusion and one graft stenosis. RESULTS: The use of intraoperative angiography improved the patency rate from 90 to 98% and reduced the incidence of perioperative stroke from 13 to 9.5%. Ninety-two percent of the patients were in excellent or good neurological condition at the time of discharge from the hospital, compared with 95% before surgery. The perioperative mortality rate was 2%. Other complications included three intracranial hematomas, rupture of a vein graft in a patient with Marfan's syndrome, and five tumor resection-related problems. The long-term survival rates for patients who received grafts were excellent for patients with benign tumors, fair to poor for patients with malignant tumors, good for patients with aneurysms, and excellent for patients with ischemia. CONCLUSION: The results of saphenous vein and radial artery grafting have been greatly improved by the use of intraoperative angiography, improvements in surgical techniques, and improved perioperative treatment. PMID- 10371621 TI - The Kawase approach to retrosellar and upper clival basilar aneurysms AB - OBJECTIVE: Fifteen basilar aneurysms approached via Kawase's anterior petrosectomy were analyzed to determine parameters that could reliably predict the applicability of this approach to specific basilar aneurysms on the basis of existing imaging. METHODS: Anatomic data were gathered by studying 40 dry skulls in which measurements were taken to define the limits of the surgical window. Clinical data were obtained from the review of charts and radiographic images of 15 patients surgically treated with the Kawase approach. The data were combined to categorize basilar aneurysms according to their position in relation to bony anatomy as seen on preoperative angiograms. RESULTS: Two relevant measurements were determined on lateral angiograms that were predictive of the applicability of operative approach. The K1 line determined the caudal extent of exposure of the Kawase approach to be 18 mm below the floor of the sella turcica and represented the distance to the floor of the internal auditory meatus. The K2 line determined the caudal extent of exposure of the posterior petrosectomy approach to be 24 mm below the floor of the sella turcica and represented the distance to the upper aspect of the jugular tubercle. Basilar aneurysms below the posterior clinoid process could be categorized in relationship to the regional bony anatomy in a manner that is predictive of the appropriate surgical approach as 1) retrosellar, 2) upper clival, 3) midclival, and 4) lower clival. Glasgow outcome data in 15 patients surgically treated with the Kawase approach demonstrated results comparable to those reported for ruptured basilar aneurysms. CONCLUSION: Individual basilar artery aneurysms can be categorized according to their relationship to bony anatomy on lateral view preoperative angiograms without bone subtraction. Anatomic parameters, the K1 and K2 lines, from these angiograms enable the neurosurgeon to predict the most appropriate approach for each type of basilar artery aneurysm. PMID- 10371622 TI - Role of transcranial Doppler monitoring in the diagnosis of cerebral vasospasm after subarachnoid hemorrhage AB - OBJECTIVE: The purpose of this study was to determine the correlation between transcranial Doppler (TCD) velocities and angiographic vasospasm after aneurysmal subarachnoid hemorrhage. METHODS: In the first part of this study, patients were retrospectively reviewed to correlate middle cerebral artery absolute blood flow velocities with angiographic vasospasm. In the second part of the study, the middle cerebral artery/ipsilateral extracranial internal carotid artery velocity ratio (Lindegaard ratio) was prospectively correlated with angiographic vasospasm. Angiographic vasospasm was independently graded, by observers blinded to the TCD results, as either none, mild (less than one-third artery luminal narrowing), moderate (one-third to one-half narrowing), or severe (more than one half narrowing). The sensitivity, specificity, likelihood ratios for positive and negative TCD results, positive and negative predictive values, and kappa and P values were calculated. RESULTS: One hundred one patients were analyzed in the first part of the study, and 44 patients were analyzed in the second part. Interobserver agreement regarding angiographic vasospasm was good (kappa = 0.86). Despite significant correlation between mean velocities and the degree of vasospasm, the clinical dependability of TCD velocities (evaluated using predictive values and likelihood ratios) was limited. The positive predictive value of velocities of > or =200 cm/s for moderate/severe angiographic vasospasm was 87% but that of lower velocities was approximately 50%. The negative predictive value of velocities of <120 cm/s was 94% but that of higher velocities was approximately 75%. Only the likelihood ratios for velocities of <120 or > or =200 cm/s were useful (likelihood ratio for negative result = 0.17, likelihood ratio for positive result = 16.39). Overall, 57% of patients exhibited maximum velocities in the indeterminate range between 120 and 199 cm/s. Lindegaard ratios did not improve the predictive value of TCD monitoring. CONCLUSION: For individual patients, only low or very high middle cerebral artery flow velocities (i.e., <120 or > or =200 cm/s) reliably predicted the absence or presence of clinically significant angiographic vasospasm. Intermediate velocities, which were observed for approximately one-half of the patients, were not dependable and should be interpreted with caution. PMID- 10371623 TI - Integration of functional magnetic resonance imaging supported by magnetoencephalography in functional neuronavigation AB - OBJECTIVE: In this study, the intraoperative visualization of functional data provided by functional magnetic resonance imaging (fMRI) and magnetoencephalography (MEG) leading to functional neuronavigation is demonstrated in surgery around the motor strip. METHODS: In seven patients with lesions adjacent to the central region, fMRI was performed with a 1.5-Tesla magnetic resonance system, using axial echo-planar imaging with a motor and a sensory task. Somatosensory and motor evoked fields were recorded with a biomagnetometer. fMRI and MEG were matched to an anatomic three-dimensional magnetic resonance image set by a contour fit. Then this three-dimensional image data set was transferred to the navigation microscope and displayed in the eyepieces of the microscope during surgery. Additionally, intraoperative recording of somatosensory evoked potentials was performed for verification of the central sulcus. RESULTS: In all cases, the projection of fMRI and MEG data into the operating viewing field allowed easy identification of the central region, which was confirmed by phase reversal of somatosensory evoked potentials in each case. fMRI and MEG measurements yielded corresponding results in each patient. CONCLUSION: Functional neuronavigation with integration of fMRI and MEG allows the fast identification of eloquent brain areas. The widespread availability of fMRI will result in a broad availability of functional neuronavigation, which will, in turn, contribute to the successful surgery of lesions in eloquent brain areas with lower morbidity. PMID- 10371624 TI - Endoscopic treatment of the trapped fourth ventricle AB - OBJECTIVE: To propose endoscopic treatment as an effective initial alternative for patients with a trapped fourth ventricle. METHODS: We reviewed the records of the last 16 consecutive patients with a symptomatic trapped fourth ventricle seen at the Arkansas Children's Hospital. The first eight patients underwent a shunt procedure; the next eight had endoscopic procedures. The shunt procedures consisted of either separate shunts or combined supra- and infratentorial shunts with shared distal catheters. The endoscopic procedures consisted of either fenestration into the lateral ventricle or aqueductal plasty with or without a stent. RESULTS: All patients underwent successful procedures with good outcomes, although the patients with shunts appeared to have a higher-than-expected rate of revision (50%). Seven revisions were performed on four patients, with a complication rate of 25%. Of the patients who had endoscopic procedures, one eventually required a shunt. The overall complication rate for patients who had endoscopic procedures was also 25%. CONCLUSION: Endoscopic treatment of the trapped fourth ventricle is effective in most cases. In view of the higher-than expected revision rate with fourth ventricular shunts and an equivalent complication rate, endoscopic treatment is a reasonable initial treatment option for patients with a trapped fourth ventricle. PMID- 10371625 TI - Cranial procedures without hair removal AB - OBJECTIVE: In 1992, Winston published the first large series of patients undergoing cranial neurosurgery without hair removal (Winston KR: Hair and neurosurgery. Neurosurgery 31:320-329, 1992). Prompted by this report, the senior author began a prospective trial in 1992 of cranial neurosurgery without hair removal. METHODS: All patients undergoing elective cranial surgery were offered the opportunity to undergo surgery without hair removal. The protocol advocated by Winston was strictly followed in the first 100 patients but has subsequently been modified. Patients having only cranial procedures have their head prepared for 10 minutes with chlorhexidine and water followed by an isopropyl alcohol rinse. Patients having craniofacial procedures are prepared with iodophor. A single perioperative dose of prophylactic antibiotic is administered. RESULTS: We have performed 346 cranial operations without hair removal. These include craniotomy for tumor, trauma, and aneurysm (n = 115); epilepsy procedures, including depth, subdural strip, and grid electrode placement, lobectomy, and callosotomy (n = 95); functional procedures, including thalamotomy, pallidotomy, capsulotomy, and stereotactic biopsy (n = 84); ventriculoperitoneal shunts (n = 8); brain abscess aspiration or resection (n = 5); and miscellaneous other procedures (n = 10). Twenty-nine patients underwent cranial base procedures in conjunction with an otolaryngologist and had the alternate preparation. There have been no infections and no other complications associated with not removing hair. CONCLUSION: Cranial surgery without hair removal is safe and is not associated with a discernible increased risk of infection. There are simple techniques for keeping hair out of the wound. Patients are highly desirous of keeping their hair and react very positively to this option. We advocate a greater practice of this technique in neurosurgery. PMID- 10371626 TI - Posterior cervical arthrodesis and stabilization: an early report using a novel lateral mass screw and rod technique AB - OBJECTIVE: Posterior cervical arthrodesis and stabilization with lateral mass plates is a biomechanically sound construct in multiple planes of motion. It is reproducible and especially useful when the posterior elements are missing or fractured. Unfortunately, it is difficult to use in patients with severe degenerative spondylosis because the plate is malleable only in the sagittal plane and the screw positions are dictated by the plate's entry holes. METHODS: A novel system of lateral mass screws that can be positioned before placement of a lateral construct was used in nine patients. Their outcomes as well as the technical applications of this system were reviewed. RESULTS: A total of 52 screws were placed in nine patients who underwent posterior cervical arthrodesis with the Cervifix system (Synthes USA, Paoli, PA). Diagnoses included trauma in four patients, degenerative spondylosis in three, and tumor in two. Rods were molded individually according to the patient's anatomy. Compression, distraction, and lateral rotation, if indicated, were performed. Follow-up averaged 36 weeks. Lateral and anteroposterior radiographs, obtained at progressive intervals, revealed excellent fixation and screw purchase without pull-out. There were no cases of spinal cord, nerve root, or vertebral artery injury. CONCLUSION: The Cervifix system accommodates variation in anatomic size and spacing of the lateral masses, potentiating precise screw placement. The rods can be molded in multiple planes, and selective application of compressive, distractive, or lateral rotatory forces is allowed. The system is very straightforward and simple to use, and we have had good success without pseudarthrosis or complications from screw placement in our series. PMID- 10371627 TI - Relationship between global and cortical cerebral blood flow in patients with head injuries AB - OBJECTIVE: The goal of this study was to analyze the use of thermal diffusion cerebral blood flow (TD-CBF) monitoring as a continuous monitoring method for patients with head injuries in the intensive care unit. METHODS: The TD-CBF probe was placed on normal-appearing frontal or parietal cortex at the time of surgery to treat traumatic intracranial hematomas in 35 patients, and cortical cerebral blood flow (CBF) was monitored for up to 7 days after surgery. We compared TD-CBF values with global CBF values measured by the standard Kety-Schmidt technique, and we compared changes in TD-CBF with changes in jugular venous oxygen saturation observed during intracranial pressure elevations. RESULTS: The average value for the global CBF measurements was 50.5+/-0.9 ml/100 g/min and that for the TD-CBF measurements was 60.5+/-1.4 ml/100 g/min; the average difference was 9.3+/-1.2 ml/100 g/min. The overall slope of the regression between the global CBF and TD-CBF measurements (n = 206) was 0.636 (comparison of observed slope with a slope of 0, P < 0.001). The relationship between the TD-CBF and global CBF values during 546 episodes of increased intracranial pressure was examined by comparing the changes in TD-CBF with the changes in jugular venous oxygen saturation. When the change in TD-CBF was at least 10 ml/100 g/min during an intracranial pressure elevation, the TD-CBF change reflected the change in jugular venous oxygen saturation on 85% of the occasions. CONCLUSION: The TD-CBF method is very convenient because of the continuous and automatic nature of the measurements. Most of the time, a change in TD-CBF indicated a similar change in global CBF. However, the limitations of local measurements of CBF must be kept in mind during therapeutic decision-making. PMID- 10371628 TI - Evaluation of a new fiberoptic catheter for monitoring jugular venous oxygen saturation AB - OBJECTIVE: The purpose of this study was to evaluate three modified, fiberoptic, oxygen saturation catheters as monitors of jugular venous oxygen saturation (SjvO2). METHODS: Three modified fiberoptic catheters, designated Catheters 1, 2, and 3, were evaluated. After preinsertion calibration, the catheters were inserted in the dominant jugular vein of patients with severe head injuries. The catheter reading for SjvO2 was compared with the SjvO2 value measured in a blood sample drawn through the catheter, at intervals up to 72 hours. RESULTS: Catheter 3 (a modified version of the pediatric Swan-Ganz catheter) showed significantly better performance than the other two catheters. Both the initial calibration of the catheter and the calibration with time were significantly better for Catheter 3 than for the other two catheters tested. The Catheter 3 value for SjvO2 was more than 4% different from the co-oximeter value in only 6% of the 4-hour calibration checks, compared with 26% and 29% for Catheters 1 and 2, respectively. CONCLUSION: Catheter 3, which is now being marketed as the Opticath P540-H catheter, represents a significant improvement in performance and may make SjvO2 monitoring in the intensive care unit more practical. PMID- 10371629 TI - Why have recent trials of neuroprotective agents in head injury failed to show convincing efficacy? A pragmatic analysis and theoretical considerations AB - An overview of the results of recent trials of neuroprotective agents in head injury is presented. None of the trials showed efficacy in the general population of patients with a severe head injury. A critical analysis of the possible reasons for this failure is given. Specific attention is focused on the heterogeneity of the patient population, the importance of baseline prognostic indicators, and the problems caused by the distribution of outcome and the dichotomization of these outcomes in the Glasgow Outcome Scale. Recommendations are presented for consideration in the design and analysis of future trials in head injury. PMID- 10371630 TI - Image-guided robotic radiosurgery AB - PURPOSE: To describe the design and performance of a novel frameless system for radiosurgery. This technology, called image-guided radiosurgery (IGR), eliminates the need for stereotactic frame fixation by relating the identified lesion to radiographic landmarks. CONCEPT: IGR uses a lightweight x-band linear accelerator, computer-controlled robotic arm (Fanuc manipulator [Fanuc Robotics North America, Inc., Rochester Hills, MI]), paired orthogonal x-ray imagers, and a computer workstation that performs rapid image-to-image registration. During radiosurgery, the x-ray imaging system determines the location of the lesion and communicates these coordinates to the robot, which adjusts the pointing of the linear accelerator beam to maintain alignment with the target. RATIONALE: Existing stereotactic techniques require rigid cranial fixation to establish and maintain a system of reference for targeting. Such frames cause pain for the patient, limit the use of fractionation, and necessitate a prolonged period of general anesthesia if children are to be treated. Furthermore, skeletal or any other type of rigid fixation is difficult to achieve beyond the cranium. IGR was designed to overcome these limitations, which are inherent to nearly all current radiosurgical methods. DISCUSSION: Preliminary testing and early clinical experience have demonstrated the practicality and potential of the IGR concept and have identified the most important directions for improvement. For example, an IGR prototype accurately tracked target displacements in three dimensions but showed reduced accuracy when confronted by rotational movements. This observation led to development of a new generation of tracking algorithm that promises to improve tracking in all six dimensions. Further experience indicated that improvements in the quality of the x-ray images were needed to allow the system to locate and treat target sites outside the cranium. Consequently, a new x-ray imaging technology with superior resolution and increased sensitivity has been added to the system. These improvements should make it possible to apply IGR techniques to a variety of targets located throughout the body. This article describes and critiques the components of the IGR and summarizes our preliminary clinical experience. PMID- 10371631 TI - Anterior choroidal artery supply to the posterior cerebral artery distribution: embryological basis and clinical implications AB - OBJECTIVE AND IMPORTANCE: We report four cases of anomalous anterior choroidal artery supply to the temporal, parietal, and occipital cortical regions normally supplied by branches of the posterior cerebral artery. Three of these cases were associated with significant intracranial vascular pathology. We examine the embryological basis for the anomalous vascular anatomy to emphasize the potential variability of the anterior choroidal artery distribution and illustrate the spectrum of this variability angiographically. These variants may also have significant implications for surgical and endovascular treatment. CLINICAL PRESENTATION: One patient was diagnosed with an arteriovenous malformation, and two patients were diagnosed with aneurysms, one of which presented with a subarachnoid hemorrhage. INTERVENTION: The patient with the arteriovenous malformation underwent preoperative embolization and surgical resection, one patient with an aneurysm was treated via surgical clipping, and the other aneurysm underwent endovascular coiling. CONCLUSION: This group of patients illustrates an underemphasized but potentially important vascular anomaly. Phylogeny and embryology of the intracerebral vessels suggest a mechanism related to transposed distributions. Careful attention must be directed to the microvascular anatomy of the intracerebral circulation so that these anomalies are recognized and, if necessary, factored into surgical and interventional planning. PMID- 10371632 TI - Infraoptic course of the anterior cerebral artery associated with an anterior communicating artery aneurysm: anatomic case report and embryological considerations AB - OBJECTIVE AND IMPORTANCE: An infraoptic course of the proximal anterior cerebral artery is a rare anomaly that has been reported in 32 cases to date, often in association with cerebral aneurysms. This anomaly represents a maldevelopment in the embryogenesis of the anterior circle of Willis, resulting from the persistence of the primitive prechiasmal arterial anastomosis or an error in the development of the definitive ophthalmic artery (OphA). The case of a patient with a ruptured middle cerebral artery aneurysm and an anterior communicating artery aneurysm associated with this anomaly is described, and the anatomic features are outlined. CLINICAL PRESENTATION: A 30-year-old male patient with a right temporal hematoma and subarachnoid hemorrhage was admitted to our department 4 days after the hemorrhaging episode, with normal neurological examination results. Angiography revealed a right middle cerebral artery aneurysm and an anterior communicating artery aneurysm with an anomalous precommunicating tract. INTERVENTION: The patient was surgically treated 14 days after the hemorrhaging episode, through a right frontopterional craniotomy; both aneurysms were excluded by clipping. The anomalous infraoptic proximal tract of the anterior cerebral artery was well documented, with its origin adjacent to the OphA. The patient remained neurologically intact after surgery and was discharged 8 days later. CONCLUSION: The anomalous infraoptic course of the proximal anterior cerebral artery was associated with a low bifurcation of the ipsilateral internal carotid artery and the absence of the contralateral precommunicating tract in this patient. The strict anatomic relationship with the origin of the OphA suggests an error in the development of the definitive OphA, with persistence of an anastomotic loop between the primitive dorsal and ventral OphAs. It is concluded that, for aneurysm surgery, careful angiographic evaluation and an understanding of the neurovascular relationships in the circle of Willis are essential for a successful postoperative course, especially when very rare vascular anomalies are treated. PMID- 10371633 TI - Postoperative hypotension after carotid angioplasty and stenting: report of three cases AB - OBJECTIVE AND IMPORTANCE: Hemodynamic instability after carotid angioplasty and stenting is not well recognized. We report three patients who developed sustained hypotension in the postoperative period after successful carotid angioplasty and stent placement for internal carotid artery stenosis in the carotid sinus region. CLINICAL PRESENTATION: In two patients, hypotension was initially induced by inflation of the angioplasty balloon. The third patient developed hypotension after completion of the procedure. In all cases, the hypotension persisted for 18 to 33 hours after the procedure. During the postoperative period, two of these patients also developed sinus bradycardia, which, in one patient, was further complicated by a third-degree atrioventricular block. INTERVENTION: The hypotension was successfully treated by intravenous vasopressors or inotropic agents. No permanent neurological or cardiac sequelae were observed. CONCLUSION: Sustained hypotension with or without bradycardia may develop after carotid angioplasty and stent placement, presumably as a result of carotid sinus dysfunction. During the postoperative period, patients should be monitored in settings suited to expeditious management of cardiovascular emergencies. PMID- 10371634 TI - Posterior fossa swelling and hydrocephalus resulting from hypertensive encephalopathy: case report and review of the literature AB - OBJECTIVE AND IMPORTANCE: Brain stem and cerebellar edema rarely have been described as the principal manifestation of hypertensive encephalopathy. In addition, secondary hydrocephalus has been described in only a few cases in the literature. We present an unusual case of posterior fossa swelling and hydrocephalus resulting from hypertensive encephalopathy. CLINICAL PRESENTATION: A 53-year-old man presented with increased shortness of breath, headache, and visual changes, which had been worsening for several months. Blood pressure on presentation was 253/140 mm Hg. Neuroradiological studies revealed brain stem swelling predominantly affecting the pons, with compression of the adjacent cisterns and fourth ventricle and resultant hydrocephalus. The diagnosis of brain stem glioma was briefly entertained. INTERVENTION: The patient's blood pressure was brought under control with medical management, and he was treated with dexamethasone for 48 hours. Subsequent neuroradiological studies revealed decreased posterior fossa edema as well as marked improvement in the hydrocephalus. CONCLUSION: Hypertensive encephalopathy can present principally in the posterior fossa and can give rise to obstructive hydrocephalus. Invasive treatment of the hydrocephalus is not necessarily required in this clinical setting because reduction of the blood pressure may result in rapid improvement of the hydrocephalus. PMID- 10371635 TI - Venous angiomas with arteriovenous shunts: report of three cases and review of the literature AB - OBJECTIVE AND IMPORTANCE: In spite of recent recognition of the benign nature of venous angioma (VA), only limited information is available on the clinical features of VA with arteriovenous shunt (AVS). The purpose of this study was to elucidate the clinical profile of VA with AVS. CLINICAL PRESENTATION AND INTERVENTION: We describe three patients having a VA with AVS and review the clinical features of 31 patients reported in the literature, including our three patients. The patients included 12 women and 19 men, ranging in age from 18 to 54 years. Seven patients (22.6%) presented with intracranial hemorrhage, and none of 16 patients developed a new or recurrent hemorrhage (mean follow-up period, 11 months). Treatment was conservative in 14 patients, lobectomy or partial resection of the VA in 6, removal of hematoma in 4, operation only for coexisting aneurysm or arteriovenous malformation in 4, and not known in 3. The outcome was reported as good recovery in 19 patients, persistent neurological deficits in 2, death or deterioration not related to the VA in 3, and not known in 7. CONCLUSION: Although there remains some uncertainty as to the clinical features of VA with AVS, its prognosis seems to be essentially as benign as that of VA without AVS. Thus, conservative treatment is recommend except for patients with a large hematoma or with a coexisting arteriovenous malformation or a symptomatic, accessible cavernous angioma, which may be treated by surgical intervention. Further collection of data is required to establish definite treatment guidelines. PMID- 10371636 TI - Parasellar neurenteric cyst: unusual site and histology: case report AB - OBJECTIVE AND IMPORTANCE: Intracranial neurenteric cyst is an exceedingly rare, congenital, benign, space-occupying lesion, and only a few reports of intracranial neurenteric cysts have been documented. We report a case of parasellar neurenteric cyst, a site not previously reported in the literature. A further rarity was the presence of smooth muscle as part of the cyst wall. This feature provides support for an endodermal origin of the cyst. CLINICAL PRESENTATION: A 27-year-old man presented with recurrent episodes of headache, vomiting, and left ptosis. Magnetic resonance imaging disclosed a cystic lesion in the parasellar region. INTERVENTION: A well-circumscribed cystic lesion was resected through a left frontotemporal craniotomy. At follow-up examination, persistent cranial nerve deficits were present. CONCLUSION: Hitherto unreported, a parasellar location of a well-encapsulated neurenteric cyst is described. The presence of a subepithelial smooth muscle layer, reminiscent of bronchiolar/intestinal muscularis mucosa, provides strong evidence for an endodermal origin of this cystic lesion. PMID- 10371637 TI - Giant ventral intradural extramedullary neuroma: case report AB - OBJECTIVE AND IMPORTANCE: We report an extremely rare, giant, ventrally located, intradural extramedullary neuroma in a 32-year-old woman. It extended from the foramen magnum to the level of the 10th thoracic vertebra. To our knowledge, this is the largest reported intraspinal neuroma. CLINICAL PRESENTATION: A mixed polyradicular and medullary lesion with symptoms and signs of raised intracranial pressure was observed on clinical presentation. Magnetic resonance imaging of the entire neuraxis was suggestive of a neuroma. Histological findings revealed characteristics of a classic neuroma. INTERVENTION: A complete surgical excision of the tumor was accomplished through an osteoplastic laminotomy from C1 to T10 with monitoring of the somatosensory evoked potentials. CONCLUSION: Even such a huge, ventrally located, intradural extramedullary tumor, as in this patient, can be completely removed with good results when a preoperative radiological and histological diagnosis is considered. With the help of new-generation magnetic resonance imaging scans, the intradural neural, vascular, and other structures can be clearly outlined. These tests also obviate the need for invasive diagnostic procedures such as myelography. We recommend an osteoplastic laminotomy instead of laminectomy to reduce the risk of postoperative spinal instability. PMID- 10371638 TI - Surgical treatment of cervical spine fibrous dysplasia: technical case report and review AB - OBJECTIVES AND IMPORTANCE: Cervical spine compromise by fibrous dysplasia is rare; only 20 cases have been reported. Treatment was surgical in six cases, but the management differed among the other cases. In this retrospective case report, we analyze and discuss the surgical treatment of this condition in the context of previous reports. CLINICAL PRESENTATION: A 35 year-old woman presented 1 week after the onset of spontaneous neck and left arm pain, which increased 48 hours later. A physical examination revealed only slight neck stiffness. Her previous history was unremarkable. Plain radiographic, radioisotope bone scanning, and computed tomographic studies were performed. The body of the fifth cervical vertebra was collapsed. INTERVENTION: A C5 corpectomy was performed, and the spine was reconstructed with a block of biocompatible osteoconductive polymer and a plate with four bicortical screws. Pathologic examination revealed fibrous dysplasia. The postoperative course was uneventful, and the pain disappeared. Radiographs obtained postoperatively demonstrated good spinal alignment. The patient was discharged after 7 days. Three years later, she continued to have no pain, and spine stability was preserved. CONCLUSION: Good results can be achieved in the surgical treatment of dysplastic tissue with the use of instruments for spine reconstruction and synthetic grafting to prevent invasion. PMID- 10371639 TI - Stereotaxy for hypothalamic hamartoma with intractable gelastic seizures: technical case report AB - OBJECTIVE AND CLINICAL IMPORTANCE: We report a patient with a hypothalamic hamartoma treated by stereotactic radiofrequency thermocoagulation. CLINICAL PRESENTATION: A 15-year-old girl presented with a hypothalamic hamartoma associated with intractable gelastic and tonic seizures. There were no clinical signs of precocious puberty. Magnetic resonance imaging revealed an isointense suprasellar mass, about 1 cm in diameter. TECHNIQUE: A depth electrode was placed into the hamartoma using a computed tomography-compatible stereotactic frame. Depth electroencephalographic studies allowed us to record the seizure onset from the lesion, and stereotactic radiofrequency thermocoagulation produced seizure remission. CONCLUSION: These findings suggest that hypothalamic hamartoma itself has intrinsic epileptogenicity. We believe that this surgical treatment is effective for the relatively small hypothalamic hamartoma associated with intractable gelastic seizures. PMID- 10371640 TI - Handgrip-detachable soft tissue retractor for microsurgery: technical note AB - OBJECTIVE: We developed a modified soft tissue retractor for preventing problems that occur outside of the microscopic view field during microsurgery. INSTRUMENTATION AND OPERATIVE PROCEDURE: The characteristics of the modified soft tissue retractor are detachable handgrips, which can be easily released and fixed with the manipulation of an equipped button, and a new lock adjuster, which changes its form and position so that it is withdrawn into a corner space between both arms in proportion to the open angle of the arms. The retractor is used with handgrips, as in the conventional method, until the microsurgery begins. When microsurgery begins, the handgrips are released. We have used this retractor in 45 microsurgeries and 42 burr-hole surgeries, such as stereotactic or endoscopic operations. RESULTS: The detachable handgrips can resolve problems caused by handgrips and a lock lever in conventional retractors, such as interference with the free movement of the microsurgical operator's hands, restriction of operative approach angle or direction of the light axis of a microscope, and catching or tangling of instrument cords and thin thread. In addition to microsurgery, it was beneficial in burr-hole surgeries, because operators can manipulate instruments, such as a stereotactic arm or an endoscope, without having to handle the retractor handgrips. CONCLUSION: Our modified soft tissue retractor is very useful for performing steady and safe microsurgery. PMID- 10371641 TI - Instrumentation, technique, and technology. Axon Guideline System 3000 PMID- 10371642 TI - Abducens nerve palsy after radiofrequency rhizolysis for trigeminal neuralgia: case report PMID- 10371644 TI - Nonneoplastic intramedullary spinal cord lesions mimicking tumors PMID- 10371643 TI - Asymptomatic familial cerebral aneurysms PMID- 10371645 TI - Alignment correction algorithm for transformation of stereotactic anterior commissure/posterior commissure-based coordinates for image-guided functional neurosurgery PMID- 10371646 TI - Failure of hydroxyapatite cement to set in repair of a cranial defect: case report PMID- 10371647 TI - Cocaine preexposure fails to sensitize the acquisition of cocaine-induced taste aversions. AB - In two separate experiments, rats were given either an intraperitoneal (IP) injection of 10 mg/kg cocaine once a day for 10 consecutive days (Experiment 1) or a single IP injection of 40 mg/kg of cocaine (Experiment 2) prior to receiving repeated pairings of a novel saccharin solution with cocaine (32 mg/ kg; subcutaneous; SC). Although vehicle-preexposed subjects given saccharin-cocaine pairings readily acquired an aversion to the cocaine-associated saccharin solution, subjects preexposed to cocaine (whether 10 times or only once) displayed a retarded acquisition of the aversion. That is, cocaine preexposure attenuated the acquisition of cocaine-induced taste aversions. There was no difference in the degree of attenuation between the two preexposure conditions. Thus, under conditions that are effective in inducing sensitization within other behavioral preparations there was no evidence of sensitized cocaine-induced taste aversions. The results from the present investigation are similar to reports from this laboratory and others demonstrating that preexposure to cocaine, as with a range of other psychoactive drugs, results in weaker taste aversions. The basis for the attenuating effects of cocaine preexposure was discussed in terms of an adaptation to the aversive effects of cocaine. PMID- 10371648 TI - The dopamine D3/D2 receptor agonist 7-OH-DPAT induces cognitive impairment in the marmoset. AB - Previous work has shown that dopaminergic systems are involved in cognitive function in the common marmoset. The present study investigated the role of dopamine D3 receptors in cognitive performance in the marmoset. The effects of the putative dopamine D3 receptor agonist, 7-OH-DPAT, on performance of a same day reversal visual object discrimination task were assessed using a miniature Wisconsin General Test Apparatus (WGTA). Within the same test session marmosets acquired a two-choice object discrimination initial task and a reversal task to criterion. 7-OH-DPAT (6-10 microg/kg) significantly impaired reversal task performance only, without affecting acquisition of the initial task. A higher dose of 25 microg/kg 7-OH-DPAT impaired initial task acquisition as well as reversal task acquisition, possibly as a consequence of a nonspecific influence on motor function. The dopamine D2 receptor antagonist (-)sulpiride (5-10 microg/kg) and the alpha2-receptor antagonist yohimbine (50 microg/kg) failed to attenuate the effects of 7-OH-DPAT (6 microg/kg) in this task. In contrast, the dopamine D2/D3 receptor antagonist raclopride (50 microg/kg) significantly attenuated the 7-OH-DPAT-induced impairment of reversal task performance. These results suggest that activation of dopamine D3 receptors produces a selective impairment of aspects of cognitive function in the marmoset. PMID- 10371649 TI - Effects of dopaminergic agents and of an NMDA receptor antagonist on motor coordination in Lurcher mutant mice. AB - Lurcher mutant mice, characterized by an ataxic gait and olivocerebellar degeneration, were evaluated for motor coordination in the coat-hanger test after peripheral injections of two doses of dextromethorphan, a noncompetitive N-methyl D-aspartate receptor antagonist, L-dopa/carbidopa, and SKF 77434, a dopamine D1 receptor agonist. There was an improvement in the distance traveled on the suspended horizontal string after 25 and 50 mg/kg of dextromethorphan and 37.5 mg/kg of L-dopa/carbidopa, but not after SKF 77434. None of the drugs reduced movement times or increased latencies before falling. These results indicate that NMDA receptor antagonism or stimulation of some dopaminergic mechanisms partially improve genetically determined cerebellar ataxia in mice. PMID- 10371650 TI - Combined action of thioperamide plus scopolamine, diphenhydramine, or methysergide on memory in mice. AB - The aim of the present experiments was to test the role played by the interaction of the selective H3 receptor antagonist, thioperamide, with the cholinergic, histaminergic, and serotonergic systems in modifying memory. The behavioral tests used (open-field and passive-avoidance repetition) were selected on the basis of the action displayed by thioperamide in these behavioral situations. Posttrial administration of thioperamide (5 mg/kg) resulted in an improvement in memory consolidation, as tested in the repetition of the open-field test, but repeated posttrial administration of thioperamide (2 or 5 mg/kg) had no effect in the repetition of passive avoidance test. Scopolamine (2 mg/kg) caused a deterioration in the memory processes in both tests: this effect was blocked by 2 mg/kg of thioperamide, which was itself ineffective in the test. These results may suggest that both the improvement in memory due to thioperamide and its antagonism of the amnestic effects of scopolamine are determined by activation of central cholinergic systems, due to thioperamide inhibition of H3 heteroreceptors. Diphenhydramine (2 or 10 mg/kg) was itself ineffective in the tests, but counteracted the memory improvement caused by thioperamide in the repetition of the open-field test. The effect of diphenhydramine is discussed in terms of interactions between histaminergic and cholinergic systems. Methysergide counteracted the effect of thioperamide in the open-field test only at a high dosage (50 mg/kg). The possible implication of serotonergic systems on the effects of the methysergide-thioperamide interaction in the memory process is discussed. PMID- 10371651 TI - Effects of haloperidol on communicative and aggressive behavior in male mice with different experiences of aggression. AB - Effects of two doses of haloperidol (0.1 and 0.4 mg/kg, 30 min and 24 h, IP) on communicative and aggressive behavior in C57BL/6J male mice have been studied. Some of the mice were without prior experience of aggression ("recruits"); the others had been victorious in 20 daily aggressive confrontations ("experienced winners"). Communicative behavior was estimated as the behavioral reaction to a standard tester (loser) in the partition test. Haloperidol in either dose significantly reduced communicative behavior in the "recruits." but not in the "experienced winners." Significantly fewer attacks, less total attacking time, and total time of aggressive behavior (aggressive grooming + attacks) were demonstrated by the "experiences winners," than by the "recruits," while the latency of the first attack, the number, the total and average duration of aggressive grooming events were significantly higher. In the "recruits," haloperidol dose dependently increased the latency and decreased the number of attacks, the total attacking time, and the total time of aggressive behavior 30 min and 24 h after injection. However, haloperidol did not affect the average or total time of aggressive grooming. Neither dose significantly affected any measure of aggressive behavior in the "experienced winners." It has been concluded that repeated aggression experience reduces the pharmacological sensitivity of the dopamine receptors. PMID- 10371652 TI - 5-HT2 receptor antagonism reduces hyperactivity induced by amphetamine, cocaine, and MK-801 but not D1 agonist C-APB. AB - The hyperlocomotion induced by the noncompetitive NMDA antagonist MK-801 (0.3 mg/kg SC) in mice was attenuated by the nonselective 5-HT2 antagonist ritanserin (0.12 and 0.25 mg/kg SC) and by the 5-HT2A selective antagonist MDL100907 (0.05 and 0.1 mg/kg SC). SB242084 (0.25-1.0 mg/kg), a selective 5-HT(2C) antagonist, had no effect on MK-801-induced hyperactivity. These same doses of ritanserin and MDL100907 reduced the hyperactivity induced by cocaine (10 mg/ kg). Amphetamine (2.5 mg/kg SC) induced hyperlocomotion that was also attenuated by ritanserin (0.064).25 mg/kg SC). The hyperlocomotion induced by the D1 agonist C-APB (1.0 mg/kg) is not altered by pretreatment with ritanserin or MDL100907. This suggests that compounds that increase locomotor activity via indirectly increasing dopaminergic activity (either by increased release or blockade of reuptake) require the activation of a 5-HT2A receptor. Activity of compounds that act directly at the postsynaptic dopamine receptors such as C-APB is not dependent on such a mechanism. This suggests a selective involvement of 5-HT2A receptors but not 5-HT2c receptors in the mediation of the behavioral effects of compounds that increase synaptic concentration of dopamine but not directly acting agonists. This implies that the 5-HT2A receptors modulate elevation of extracellular dopamine, not the postsynaptic sensitivity of dopamine neurons. PMID- 10371653 TI - Involvement of opioid mu1-receptors in opioid-induced acceleration of striatal and limbic dopaminergic transmission. AB - The role of mu1-opioid receptors in the acceleration of cerebral dopaminergic transmission induced by morphine and the putative mu1-opioid agonist, etonitazene, was studied in rats by measuring the tissue levels of dopamine (DA) and its metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in the dorsal striatum and nucleus accumbens. The striatal extracellular concentrations of DA and its metabolites in freely moving rats were estimated as well. Morphine (3 mg/kg) and etonitazene (2.5 microg/kg) increased the striatal and accumbal dopamine metabolism as measured by the tissue ratios of DOPAC/DA and HVA/DA. The mu1-opioid receptor antagonist, naloxonazine (15 mg/kg), significantly antagonized these elevations except the morphine-induced elevation of striatal HVA/DA ratio. Both morphine (3 mg/kg) and etonitazene (1, 2.5, and 5 microg/kg) elevated the striatal extracellular DA, DOPAC, and HVA. Naloxonazine antagonized the effects of morphine and etonitazene on striatal extracellular DA concentration as well as etonitazene's effects on DOPAC and HVA, but not morphine's effects on DOPAC and HVA. As we previously showed concerning morphine, the conditioned place preference induced by etonitazene was inhibited by naloxonazine. These findings emphasize the role of mu1-opioid receptors in opioid reward, in which the mesolimbic dopaminergic system is considered to be importantly involved. Our results clearly show that in addition to the mesolimbic dopaminergic system the mu1-opioid receptors are also involved in the control of nigrostriatal DA release and metabolism. However, the effects of etonitazene on the striatal DA differ from those of morphine, suggesting that the opioid mechanisms regulating these two DA systems differ. PMID- 10371654 TI - Transdermal nicotine: single dose effects on mood, EEG, performance, and event related potentials. AB - A 21-mg dose of nicotine was administered transdermally to 16 overnight smoking deprived smokers in a double-blind, placebo-controlled design. Mood ratings, electroencephalography (EEG), behavioral performance and event-related potential (ERP: P300) indices of attention and information processing speed were assessed before and 4 h after placebo/nicotine treatment. Although nicotine, relative to placebo, failed to alter mood, it increased absolute and relative power indices of EEG arousal, shortened reaction times, and increased P300 amplitudes. The results are discussed in relation to nicotine's actions on cholinergic transmission and its role in smoking behavior. PMID- 10371655 TI - Sensitization of stereotyped behavior to amphetamine is context and response dependent. AB - The present study was designed to determine whether the environmental context in which amphetamine is administered plays a role in the development of sensitization to the stereotyped behavioral effects of amphetamine in mice. In male CF-1 mice, the dose-response curve for stereotyped behavior elicited by amphetamine was shifted 1.9-fold to the left 48 h after pretreatment with 14 mg/kg amphetamine. Behavioral sensitization only developed in mice that were pretreated in the same or a similar environment as that of the test environment. In addition, when mice were placed in an environment that attenuated the acute expression of stereotyped behavior elicited by the pretreatment dose of amphetamine, sensitization never developed. A further experiment showed that 96% of the mice that expressed stereotypy after the ED50 pretreatment dose of 10 mg/kg amphetamine showed a stereotyped behavioral response to the lesser dose of 7 mg/kg 48 h later, indicating sensitization. In contrast, mice that did not express stereotypy after the ED50 dose of amphetamine failed to show a significant stereotyped behavioral response to amphetamine challenge compared to vehicle-pretreated controls. Therefore, the results indicate that preexposure to a single high dose of amphetamine produces context- and response-dependent sensitization to amphetamine-induced stereotyped behavior. PMID- 10371656 TI - Orphanin FQ and behavioral sensitization to cocaine. AB - The recently identified endogenous ligand for the ORL-1 (opioid receptor-like) receptor, orphanin FQ, has been shown to induce hypolocomotion and to decrease extracellular dopamine levels in the nucleus accumbens (N.Acc) after intraventricular (ICV) administration. This study investigated the effect of intraventral tegmental area (VTA) administration of orphanin FQ on the hyperlocomotor effects of peripheral cocaine administration and on the development of behavioral sensitization to cocaine. The administration of cocaine (40 mg/kg IP) once daily for 3 days to male Sprague-Dawley rats resulted in an enhanced locomotor response to a subsequent challenge of cocaine (10 mg/kg IP) 5 days later. The bilateral administration of orphanin FQ (10 microg/side or 30 microg/side) into the VTA 5-10 min prior to the administration of cocaine (40 mg/kg IP) produced a transient (15-30 min) decrease in the hyperlocomotor response to cocaine on day 1 but not on days 2 and 3 of the sensitization paradigm. Such orphanin FQ pretreatment on days 1-3 had no effect on the development of a sensitized response to cocaine (10 mg/kg IP) 5-7 days after the last orphanin FQ injection. However, repeated intra-VTA administration of orphanin FQ (30 microg/side) alone for 3 days resulted in a sensitized response to a single dose of cocaine (10 mg/kg IP) given 5-7 days later. These results indicate that orphanin FQ decreases the activity of mesolimbic dopamine neurons via an action in the VTA, an effect that is both transient and demonstrates rapid tolerance, and consequently, is insufficient to prevent the development of cocaine sensitization. The ability of the peptide to induce cocaine sensitization when administered alone despite its acute inhibitory effects is unique and requires further study to elucidate the mechanisms responsible. PMID- 10371657 TI - Serotonin2A receptor modulation of D1 dopamine receptor-mediated grooming behavior. AB - We have previously observed that intracerebroventricular infusion of a 5-HT2A receptor antisense oligonucleotide for 8 days results in an increase in cortical 5-HT2A receptor sites and an increase in central 5-HT2A receptor function as measured by quantitation of 5-HT2A receptor-mediated headshake behavior (28). Because lesioning serotonergic neurons or chronic administration of 5-HT2A receptor antagonists does not result in an increase in 5-HT2A receptor density or function in the brain, we have taken advantage of this unique upregulation of 5 HT2A receptors following 5-HT2A receptor antisense oligonucleotide infusion to study the modulation of D1 receptor-mediated behaviors by 5-HT2A receptors. Grooming behavior, elicited by acute injection of SKF 38393, was attenuated after chronic ICV infusion of a 5-HT2A receptor antisense oligonucleotide. There was also a reduction in vacuous chewing behavior induced by SKF 38393, which did not reach statistical significance. Other oral behaviors (i.e., tongue protrusions and gnawing at the cage bottom) were not attenuated. An increase in the density of cortical, as well as striatal 5-HT2A receptor sites was observed following chronic antisense oligonucleotide administration. There was no change in striatal D1 dopamine receptors following 5-HT2A receptor antisense oligonucleotide administration. SKF 38393-induced grooming behavior was also attenuated in naive rats pretreated acutely with the 5-HT2 receptor agonist DOI. These results suggest a role for the 5-HT2A receptor in the modulation of D1 receptor function. PMID- 10371658 TI - Although chemically related to amineptine, the antidepressant tianeptine is not a dopamine uptake inhibitor. AB - We investigated whether the antidepressant tianeptine shares the dopamine uptake inhibitory properties of the chemically related antidepressant amineptine. Tianeptine dose dependently (5, 10, 20, 40 mg/kg IP) increased locomotor activity in mice. This stimulant effect (20 mg/kg IP) was dose dependently prevented not only by the D1 dopamine receptor antagonist SCH 23390 (7.5. 15, 30 microg/kg SC), but also by the D2 dopamine receptor antagonist haloperidol (50, 100, 200 microg/kg IP), in contrast to that elicited by dopamine uptake inhibitors. Where the latter prevent dexamphetamine-induced (3 mg/kg SC) reversion of akinesia in mice pretreated with reserpine (4 mg/kg SC, 5 h before test), tianeptine (20 mg/kg IP, 30 min before test) did not. Tested up to a concentration of 10-4 M, tianeptine did neither inhibit the [3H]dopamine uptake into mouse striatal synaptosomes nor compete in vitro with the specific binding of [3H]WIN 35,428 at dopamine transporters from striatal membranes. Finally, in mice injected IV with a tracer dose of [3H]WIN 35,428 (1 microCi), the highest tested dose of tianeptine (40 mg/kg IP) did not reduce the specific binding of the radioligand to striatal dopamine transporters. It is concluded that the antidepressant effect of tianeptine does not depend upon a blockade of the neuronal dopamine transporter. PMID- 10371659 TI - Electrophysiological response to neuropeptide Y (NPY): in alcohol-naive preferring and non-preferring rats. AB - Electroencephalograms (EEGs) and event-related potentials (ERPs) to auditory stimuli were recorded following intracerebroventricular administration of neuropeptide Y (saline, NPY: 1.0, 3.0 nmol) in two lines of rats that have been genetically selected for alcohol preferring (P) or non-preferring (NP) behaviors. Previous studies have demonstrated that NPY has a distinct electrophysiological profile that is similar to that of ethanol. In outbred Wistar rats, both NPY and ethanol produced highly significant decreases in the amplitude and increases in the latency of the N1 component of the ERP to all three auditory stimuli. Because the N1 has been associated with attention, these data suggest that both NPY and alcohol may diminish attentional processes. In the present study, NPY-induced decreases in N1 amplitude were also found, but only to the frequently presented tone. This suggests that both P and NP rats may have attenuated responses to NPY's effects on attention/arousal. Like outbred Wistars, P and NP rats were also found to have significant NPY-induced increases in N1 latency in the cortex and hippocampus. However, in the amygdala, while P rats evidenced increases in N1 latency and decreases in N1 amplitudes, NP rats displayed the opposite effects. Spectral analysis revealed that NPY also produced differential EEG responses in P and NP rats. In previous studies in outbred Wistar rats NPY has been found to produce slowing of delta (1-2 Hz) frequencies at the 1-nmol dose and reductions in power, particularly in the higher frequencies in the amygdala, at the 3-nmol dose. This electrophysiological profile is not unlike what is seen following alcohol and benzodiazepines and is associated with anxiolysis. P rats were found to have this general pattern of EEG responses to NPY but attenuated suggesting that they may have reduced responses to electrophysiological measures of the anxiolytic effects of NPY. In contrast, NP rats had NPY-induced EEG effects in amygdala and frontal cortex that were opposite to those seen in P rats. These opposing responses to NPY tended to produce a "normalization" of the power differences that existed between the two rat lines at baseline. Taken together with previous findings that P rats have decreased NPY concentrations in limbic and frontal cortical sites, these data suggest that differences in the regulation of NPY neurons may contribute to the expression of behavioral preference for ethanol consumption in these rat lines. PMID- 10371660 TI - Reinforcement value of gustatory stimuli determined by progressive ratio performance. AB - The progressive ratio schedule provides a means to determine reinforcement value that is independent of response rate. This is achieved by increasing the number of lever presses for each successive reinforcement until, eventually, the subject fails to respond within a designated time limit. The number of responses in the final completed ratio defines the break point. When tested with appetitive gustatory stimuli (sucrose and saccharin), rats showed a concentration-dependent increase in break point when food deprived (Experiment 1A and, for sucrose only, Experiments 1B and IC) but not when water deprived (Experiment 1A). For aversive gustatory stimuli (sodium chloride, citric acid, and quinine) break point declined as concentration increased (Experiment 1A). In Experiment 1C, reinforcement duration had a significant, although relatively small, influence on break point for each of three concentrations of sucrose. Experiment 2 found a dose-dependent effect of the neuroleptic drug haloperidol (0.025, 0.05, and 0.1 mg/kg) on break point for sucrose reinforcement. The present study demonstrates that the progressive ratio schedule provides a valuable method to assess the influence of manipulations that might affect the perceived reinforcement value of gustatory stimuli. PMID- 10371661 TI - New techniques in stereotaxic surgery and anesthesia in the mouse. AB - Mice are used in increasing numbers in neuroscience research. This increase is linked to the availability of numerous pure genetic lines and the advent of transgenic animals. Many neuroscience techniques can be used in the mouse with success, including stereotaxic placement of cannulae and electrodes. With the recent publication of a mouse brain atlas by Franklin and Paxinos and improvements in surgical procedures for the mouse, stereotaxic surgery in mice can be performed routinely and with accuracy. In the present article, we describe techniques and apparatuses for the surgical implantation of cannulae in the mouse brain. We also present new developments in anesthesia, pain management, and postoperative care that improve survival and recovery times of mice. Using these new techniques, we have gained shorter training time for students, lower mortality rates following surgery, and faster recovery. PMID- 10371662 TI - Effects of antidepressants in rats trained to discriminate the beta-2 adrenergic agonist clenbuterol. AB - The ability of indirectly acting agonists such as norepinephrine uptake inhibitors, serotonin reuptake inhibitors, and atypical antidepressants to substitute for clenbuterol, a beta-2 adrenergic agonist, was examined in rats trained to discriminate 0.03 mg/kg clenbuterol and saline using a fixed-ratio 10 (FR 10) schedule with water reinforcement. The beta-2 selective adrenergic agonist clenbuterol produced an orderly dose-response relationship, and its discriminative stimulus effects were antagonized by the beta-adrenergic antagonist propranolol. It was found that the effects of tricyclic antidepressants and selective norepinephrine uptake inhibitors did not generalize to the discriminative stimulus effects of clenbuterol, with the exception of high doses of protriptyline. Moreover, compounds from other drug classes, including fluoxetine and phenelzine, did not substitute for clenbuterol. Atypical antidepressants, such as trazodone, rolipram, and bupropion also did not engender drug-appropriate responding. Prenalterol and dobutamine, both purported to be beta-1 adrenergic receptor agonists, partially substituted for clenbuterol, but at relatively high doses. The present results show that the antidepressants tested do not share discriminative stimulus effects with clenbuterol, a beta-2 adrenergic agonist. PMID- 10371663 TI - Baclofen alters ethanol-stimulated activity but not conditioned place preference or taste aversion in mice. AB - The present experiments examined the effects of the GABA(B) receptor agonist, baclofen, on the acquisition of ethanol-induced conditioned place preference (CPP) and conditioned taste aversion (CTA) in male DBA/2J mice. Mice in the CPP experiment received four pairings of ethanol (2g/kg) with a distinctive floor stimulus for a 5-min conditioning session (CS+ sessions). On intervening days (CS sessions), mice received saline injections paired with a different floor type. On CS+ days, mice also received one of four doses of baclofen (0.0. 2.5, 5.0, or 7.5 mg/kg) 15 min before an injection of ethanol. For the preference test, all mice received saline injections, and were placed on a half-grid and half-hole floor for a 60-min session. Baclofen dose dependently reduced ethanol-stimulated activity, but did not alter the magnitude of ethanol-induced CPP at any dose. For the CTA experiment, mice were adapted to a 2-h per day water restriction regimen followed by five conditioning trials every 48 h. During conditioning trials, subjects received an injection of saline or baclofen (2.0 and 6.0 mg/kg) 15 min before injection of 2 g/kg ethanol or saline following 1-h access to a saccharin solution. Baclofen did not alter the magnitude of ethanol-induced CTA at any dose. In addition, baclofen alone did not produce a CTA. Overall, these studies show that activation of GABA(B) receptors with baclofen reduces ethanol-induced locomotor activation, but does not alter ethanol's rewarding or aversive effects in the CPP and CTA paradigms in DBA/2J mice. PMID- 10371664 TI - Onset of the effects of the 5-HT1A antagonist, WAY-100635, alone, and in combination with paroxetine, on olfactory bulbectomy and 8-OH-DPAT-induced changes in the rat. AB - 5-HT1A receptor antagonists have recently been shown to accelerate the effects of some antidepressant drugs in clinical trials. In this study we investigate the effects of combining a full antagonist at the 5-HT1A receptor, WAY 100635 (0.2 mg/kg, SC) with the selective serotonin reuptake inhibitor (SSRI) paroxetine (5 mg/kg. SC) in the olfactory bulbectomized (OB) rat, an animal model of chronic (but not acute) antidepressant activity. Ambulation scores were measured in the open-field apparatus, following 3, 7, and 14 days of treatment. Further to the OB study, we simultaneously studied adaptive changes in 5-HT1A receptor function, utilizing alterations in the hypothermic response to the 5-HT1A receptor agonist 8-OH-DPAT. Paroxetine, in combination with WAY 100635, attenuated the hypothermic effects of 8-OH-DPAT as early as 3 days, with a full reversal evident following 7 days, whereas paroxetine, although attenuating the hypothermic effects in OB group by day 7, only reversed it fully after 14 days. Paroxetine alone and in combination with the antagonist reversed the olfactory bulbectomy-induced hyperactivity in the open field following 14 days of treatment only, this being the normal time of an "antidepressant" response in this model. However, there was no significant attenuation at any of the earlier time points. This further demonstrates that the reversal of this aspect of the olfactory bulbectomy-induced behavioral syndrome is insensitive to the potential faster onset of antidepressant action induced by 5-HT1A receptor antagonists. Nonetheless, WAY 100635, unlike previous studies with pindolol, did not interfere with the effects of the antidepressant in the model. The ability of the combination group to attenuate the hypothermic effects of 8-OH-DPAT faster than paroxetine alone, further emphasizes the role of the 5-HT1A receptor in the mechanism of action of antidepressants, and as a target for the development of faster acting antidepressants. PMID- 10371666 TI - Why is the efficacy of HAART so durable? PMID- 10371665 TI - Evaluation of intraarticular opioid analgesia for the relief of articular pain in the domestic fowl. AB - An experimental paradigm, based on the microcrystalline sodium urate-induced arthritis pain model, was used to investigate the potential peripheral analgesic properties of a variety of opioid agonists. The response criteria were changes in behavioral profiles and pain-related behaviors over 60 min commencing 1 h after intraarticular injection. The testing system was used to determine the potential optimum dose of intraarticular application of morphine sulphate (1-3 mg), fentanyl citrate (0.5-3 mg), and buprenorphine hydrochloride (0.05-1 mg). None of the opioid analgesics used had any effect on pain behavior, and it was concluded that opioids with a high affinity for the mu receptor when injected intraarticularly were unlikely to be of use in the treatment or diagnosis of inflammatory arthritic pain in the strain of domestic fowl chosen. PMID- 10371667 TI - Infectious pulmonary disease in patients receiving immunosuppressive therapy for organ transplantation. 1964. PMID- 10371668 TI - High risk genital papillomavirus infections are spread vertically. AB - It is well recognised that high-risk human papillomaviruses (HPVs) are spread by sexual activity, but the possibility of non-sexual transmission remains controversial. We present evidence for vertical transmission from at least 30% HPV positive mothers to their infants, resulting in persistent infection in children. That the mother is the source of infant infection has been confirmed by DNA sequencing. We also discuss the evidence for oral HPV-16 infection in children. In our own studies, HPV-16 DNA was detected in buccal cells from 48% children, aged 3-11 and transcriptionally active infection was confirmed in some children. Other studies have reported prevalences of 19%-27% among children less than 11 years of age. Studies that have failed to detect high-risk HPVs in children have used techniques which were insufficiently sensitive to detect the low levels of virus present. Serological studies also suggest that < or = 45% prepubertal children have acquired HPV-16. Thus, convincing evidence is now available for vertical transmission of high risk HPVs, which probably results in widespread infection among children. The consequences of such infections remain to be elucidated. PMID- 10371669 TI - High risk genital papillomavirus infections are not spread vertically. AB - The Medline-indexed literature on risk factors for HPV infection and HPV transmission is critically reviewed. Principles for assay validation and interpretation, reliability of different study designs and principles for interpretation of conflicting reports are discussed. The conclusions arrived at can be summarised as: (1) There is overwhelming epidemiological evidence that the only quantitatively important mode of transmission of infection with oncogenic genital HPV types is sexual. (2) There is also evidence that benign genital HPV types can be transmitted sexually, but the epidemiological data on the benign virus types are less extensive and less clear. (3) Perinatal HPV transmission is unequivocally demonstrated only for the rare disease juvenile respiratory papillomatosis. PMID- 10371670 TI - Regulation of DNA virus transcription by cellular POU family transcription factors. AB - In the same way as other viral functions, the transcription of viral genes is frequently controlled by cellular regulatory proteins either acting alone or together with virally encoded factors. In this review, I discuss three examples of such regulation in different types of DNA viruses by different members of the POU family of transcription factors, all of which involve viruses which play a role in the aetiology of specific human diseases. These are the glial cell specific transcription of JC virus which is controlled by the glial cell specific POU factor Tst-1; the regulation of human papillomavirus gene expression in the cervix by positively and negatively acting POU factors and the manner in which the balance between lytic or latent infection with HSV is controlled by positively and negatively acting POU factors which differ in their ability to interact with the virally encoded transactivator VP16. As well as being of interest in themselves, these processes may offer a therapeutic target for controlling the diseases caused by these very different viruses. PMID- 10371672 TI - Economic rate of discount and estimating cost benefit of viral immunisation programmes. AB - Many individual and societal decisions over purchase (or investment) involve consideration of timing, in that either the price may be paid now and the benefit enjoyed some time in the future or the converse the benefit enjoyed now and the price paid later. Since most individuals generally prefer the present to the future, economic theory has conventionally discounted future costs or benefits to estimate 'net present values'. The rationale for this is principally based on future uncertainty. In recent years, economists have turned their attention to valuing health as an economic 'good'. Observations of individual behaviour would imply that individuals discount future health, as other potential benefits, mostly because there is some uncertainty about their futures. Although economic theory is strongly predicated on the 'sovereignty of the individual', it does not necessarily follow that society discounts the future as do individuals, since for society the future is not so uncertain. Society's endorsement of many public health and preventive medicine objectives, which seek health gains in the future (rather than the present), imply that society's rate of discount may be appreciably lower than that of individuals. In immunisation, arguably one of the most effective of preventive measures, there is the additional benefit to others attributable to herd immunity. This paper argues that the future health gains for society arising from immunisation should not be underestimated by application of inappropriate discounting. PMID- 10371671 TI - Viral protein R of HIV-1. AB - Viral protein R (Vpr) of HIV-1 belongs to a class of so called 'accessory' proteins, originally thought to be dispensable for virus replication, at least in vitro. Indeed, viruses with mutated or deleted Vpr replicate well in transformed T cell lines. However, recently published results reveal several important functions performed by Vpr, which are critical for HIV-1 replication in vivo. Vpr plays an important role in regulating nuclear import of the HIV-1 pre-integration complex, and is required for virus replication in non-dividing cells. Vpr also induces cell cycle arrest in proliferating cells, stimulates virus transcription, and regulates activation and apoptosis of infected cells. These diverse functions are mediated by the interaction of Vpr with different cellular proteins, many of which carry the WxxF amino acid motif. The molecular events underlying the activity of Vpr are reviewed in this article. PMID- 10371673 TI - Simian immunodeficiency virus as a model of human HIV disease. AB - Because of strong clinical, pathological, virological and immunological analogies with HIV infection of humans, infection of macaques with SIV provides a valuable model for exploring crucial issues related to both the pathogenesis and prevention of HIV infection. The model has offered a unique setting for the preclinical evaluation of drugs, vaccines and gene-therapies against HIV, and has helped to identify many virus and host determinants of lentiviral disease. For instance, the importance of an intact nef gene for efficient lentivirus replication and disease induction, and the protective ability of live attenuated, nef-deleted viruses have been first demonstrated in macaques using molecular clones of SIV. More recently, the development of chimeric HIV-SIV vectors able to establish infection and induce disease in macaques has provided new opportunities for the evaluation of vaccination strategies based upon HIV antigens. The aim of this review is to describe the natural course of SIV infection in macaques and to outline how this model has contributed to our understanding of the complex interaction between lentiviruses and host immune system. PMID- 10371674 TI - Immunological changes associated with clinical improvement of asthmatic children subjected to psychosocial intervention. AB - In the present study we evaluated the impact of a program of psychosocial intervention (PSI) on the immunological status and the clinical management of a group of asthmatic children of an island population in Venezuela. We studied a total of 35 asthmatic children who belonged to either a PSI group (19 patients) or a control group (16 patients), both of which received conventional antiasthmatic treatment. The PSI group received, in addition, a 6-month psychosocial intervention program which included relaxation, guided imagery, and self-esteem workshops. During the PSI period, the number of asthmatic episodes and the use of bronchodilator medication were significantly reduced, and pulmonary function was significantly improved, compared to the 6 months before intervention. There was also a significant reduction in the specific IgE responses against the most important allergen in these children, the intestinal parasite Ascaris lumbricoides. PSI resulted in a significant increase of NK cells, an augmented expression of the T-cell receptor for IL-2, and a significant decrease of leukocytes with low affinity receptors for IgE. In fact, these surface markers became similar to those of nonasthmatic children from both Coche Island and the mainland. None of these clinical or immunological changes were seen in the control group of asthmatics who did not undergo PSI. These results are consistent with the possibility that PSI induces immunological alterations that are responsible for the clinical and physiological improvements observed in the study group. PMID- 10371675 TI - Cytokines inhibit sexual behavior in female rats: I. Synergistic effects of tumor necrosis factor alpha and interleukin-1. AB - The secretion of cytokines such as tumor necrosis factor alpha (TNFalpha) and interleukin-1 (IL-1) during immune activation induces sickness behavior. We have previously demonstrated that administration of either lipopolysaccharide (LPS) or IL-1 suppresses sexual behavior in female, but not in male rats. In the present study we sought to determine some of the mechanisms that are involved in mediating the alterations of female sexual behavior during immune activation. We report that sexual motivation of estrous females was reduced by intracerebroventricular administration of either recombinant rat (rr)TNFalpha (7.5 microg/rat) or rrIL-1beta (100 ng/rat), whereas sexual receptivity was altered only by IL-1beta. A significant reduction of both sexual motivation and receptivity was also induced by the combined administration of subthreshold doses of TNFalpha (3 microg/rat) and IL-1beta (20 ng/rat). These findings indicate that TNFalpha and IL-1beta act synergistically to suppress sexual motivation and receptivity. Moreover, LPS (100 microg/kg, ip)-induced reduction of sexual motivation was antagonized by the combined administration of the TNFalpha synthesis blocker pentoxifylline (50 mg/kg, ip) and IL-1 receptor antagonist (10 mg/kg, ip), but not by the administration of each of these substances by itself. In contrast, LPS-induced reduction of sexual receptivity was completely prevented by pentoxifylline. These findings indicate that the effects of LPS on sexual motivation are mediated by the synergistic effects of TNFalpha and IL-1, but only TNFalpha is required for the effect of LPS on receptivity. PMID- 10371676 TI - Cytokines inhibit sexual behavior in female rats: II. Prostaglandins mediate the suppressive effects of interleukin-1beta. AB - The proinflammatory cytokine interleukin-1 (IL-1) induces several behavioral alterations that are characteristic of illness, such as anorexia and reduced locomotor and social activity. We have recently demonstrated that IL-1 inhibits sexual activity, motivation and attractivity in female, but not in male rats following either central or peripheral administration. In the present study we examined the involvement of prostaglandin (PG) synthesis in mediating IL-1 induced suppression of female sexual behavior. Administration of the cyclooxygenase blockers indomethacin or ibuprofen completely prevented IL-1 induced suppression of female sexual behavior, including the reduction in proceptive behavior, the lordosis response to a male's mounts, and the preference for a sexually active partner. In a subsequent study, ex-vivo release of hypothalamic PGE2 and the secretion of corticosterone (CS) were measured in males and estrous females following IL-1 administration. At the same time and dose of IL-1 administration that significantly reduced sexual behavior in female but not male rats, IL-1 produced a significant increase in PGE2 release in female, but not in male rats. In contrast, IL-1 induced a significant elevation of serum CS levels in males but not in females. These findings suggest that PG synthesis is involved in mediating the effects of IL-1 on female sexual behavior. Furthermore, differential secretion of PGs and CS may underlie the gender difference in the effects of IL-1 on sexual behavior. PMID- 10371677 TI - Conditioned enhancement of antibody production is disrupted by insular cortex and amygdala but not hippocampal lesions. AB - Pavlovian conditioning procedures can be used to activate the immune system. A reliable conditioned increase of antibody production can be obtained in rats that have previously received a gustative or odor stimulus as the conditioned stimulus paired with an antigen, by exposing the animals to the conditioned stimulus alone. We showed evidence that an excitotoxic lesion bilaterally applied into the insular cortex or the amygdala, but not into the dorsal hippocampus, impaired the acquisition of both odor and gustatory conditioned immune enhancement. We found no effects of lesions on normal antibody production. These results suggest that the amygdala and the insular cortex are involved in the neural-immune interactions that mediate conditioned immunity. PMID- 10371692 TI - Journal of molecular and cellular cardiology: changing of the guard PMID- 10371678 TI - Norman Cousins Memorial Lecture 1998. Stress, personal relationships, and immune function: health implications. PMID- 10371693 TI - Extra-adrenal mineralocorticoids and cardiovascular tissue. AB - In experimental models where chronic inappropriate (relative to sodium intake and intravascular volume) elevations in circulating mineralocorticoids (aldosterone or deoxycorticosterone) are created, a reactive fibrosis with vascular remodeling is observed in systemic organs and the heart. Until recently, it was assumed that aldosterone was derived solely from adrenal glands via the circulation; however, there is now convincing evidence that cells of the heart and vasculature express genes responsible for the formation of both aldosterone and corticosterone and are capable of producing these steroids. Vascular endothelial and smooth muscle cells express CYP11B1 and CYP11B2, genes responsible for 11 beta -hydroxylase and aldosterone synthase, respectively. Furthermore, smooth muscle cells elaborate aldosterone. There is evidence that similar regulatory mechanisms operate in vascular cells as in adrenal cortex, since aldosterone synthase and 11 beta hydroxylase expression are differentially modulated by low sodium/high potassium, angiotensin II and ACTH. It is likely that such localized corticosteroid production also occurs at sites of tissue repair, where populations of collagen producing myofibroblasts, nourished by a neovasculature, predominate. Using a subcutaneous pouch model of granulation tissue we have obtained compelling data which would support such a notion. The mineralocorticoid receptor antagonist, spironolactone, severely attenuates pouch formation over a 2-week period and significantly reduces pouch wall hydroxyproline concentration. This effect is apparent even following adrenalectomy, when circulating corticosteroids are undetectable; however, with adrenalectomy alone, pouch formation is barely affected. This we took to be a possible indication of an effect of local, non adrenal steroids in maintaining pouch tissue. Spironolactone inhibits angiogenesis. A recent clinical study demonstrates the efficacy of low-dose spironolactone in enhancing survival in patients with advanced chronic cardiac failure. Although it is not known how spironolactone brings about such an improvement in survival, we would propose that inhibition of fibrous tissue formation and/or angiogenesis might be important contributory factors. Further studies are required to address the relative contributions of circulating vs local aldosterone in promoting normal vs pathologic connective tissue formation. PMID- 10371694 TI - Desensitization of cardiac beta-adrenoceptor signaling with heart failure produced by myocardial infarction in the rat. Evidence for the role of Gi but not Gs or phosphorylating proteins. AB - This study examined mechanisms of beta-adrenergic (AR) desensitization in a myocardial infarction (MI) model of heart failure in the rat. Inotropic responses to isoproterenol (non-selective beta-AR agonist) and RO 363 (selective beta1-AR agonist), in left atria and left papillary muscle, were reduced by up to 65%, while chronotropic responses in right atria were unaffected. beta1- and beta2-AR density did not change after MI, suggesting that changes in beta-AR responsiveness are due to changes occurring downstream of the receptor. Inotropic and chronotropic responses to forskolin were not altered in right and left atria and left papillary muscle after MI, suggesting changes at the level of the G proteins. Pertussis toxin treatment of animals restored inotropic responses to isoproterenol in left atria and left papillary muscle to levels seen in the sham group, indicating that inactivation of Gi-proteins improves inotropic function in MI rats, and that beta-ARs couple to Gi in cardiac failure. Expression of G protein receptor kinase 2 (GRK2), beta-arrestin1 and the regulatory subunits of cAMPdPK (RI alpha and RII alpha), showed no change after MI. However the expression of Gi alpha2 was significantly increased in left ventricle (sham 0.888+/-0.140, MI 1. 759+/-0.352 P=0.026), right ventricle (sham 0.031+/-0.004, MI 0. 037+/-0.002 P=0.006) and atria (sham 0.107+/-0.006, MI 0.138+/-0.006 P=0.004), with no changes observed in the expression of Gs alpha. These results suggest that increases in Gi play an important role in the decreased beta-AR responsiveness in the rat model of MI. PMID- 10371695 TI - Chronic colchicine administration attenuates cardiac hypertrophy in spontaneously hypertensive rats. AB - To determine whether the long-term inhibition of microtubule integrity in vivo by colchicine could attenuate the development of cardiac hypertrophy, we studied five groups of rats: Wistar-Kyoto rats receiving saline for 4 weeks (WKYsaline); WKY receiving colchicine, which depolymerizes microtubules (WKYcolchicine); spontaneously hypertensive rats receiving saline (SHRsaline); SHRs receiving colchicine (SHRcolchicine); and SHRs receiving lumicolchicine, an inactive stereoisomer of colchicine (SHRlumicolchicine). Seven-week-old animals were administered drugs or control substances via alternate day intraperitoneal injection for a period of 4 weeks. Dosage was gradually increased from 0.6 to 1.0 mg/kg to avoid drug toxicity. Depolymerization of myocardial microtubules by the in vivo administration of colchicine into the rats was confirmed by both Western blot analysis and immunofluorescence of tubulin protein in the hearts. Body weight (BW) was lower, while systolic blood pressure was significantly elevated in SHRs vs the WKY rats. No significant difference was found in either of these parameters between the control or treatment groups of each strain. Left ventricular (LV) weight-to-BW ratio was elevated and showed significant increases in the SHRs as compared to WKY animals, indicative of cardiac hypertrophy. When the SHRs were treated with colchicine but not vehicle or lumicolchicine, LV/BW was similar to the WKY. Changes of myocyte cross-sectional area determined using LV mid-free wall specimens were concordant with the LV/BW data. No significant changes were found in collagen volume fraction between groups. Thus the inhibition of microtubule polymerization abolished the progression of cardiac myocyte hypertrophy in SHRs independently of blood pressure. PMID- 10371696 TI - Length modulation of active force in rat cardiac myocytes: is titin the sensor? AB - The intrinsic cellular mechanisms by which length regulates myocardial contraction, the basis of the Frank-Starling relation, are uncertain. The aim of this work was to test the hypothesis that passive force, possibly via titin, participates in the modulation of Ca2+ sensitivity of cardiac contractile proteins induced by stretch. Titin degradation by a mild trypsin digestion modulated passive force induced by increasing from 1.9 to 2.3 microm sarcomere length in skinned rat cardiac cells. Force-pCa curves were established at these two sarcomere lengths after various durations of trypsin application that induced different passive force levels. They allowed us to evaluate myofilament Ca2+ sensitivity by the pCa of half-maximal activation (pCa50). In control conditions, stretching cells from 1.9 to 2.3 microm induced a leftward shift of pCa50 (DeltapCa50) of 0.39+/-0.03 pCa units (mean+/-SEM, n=8 cells), reflecting an increase in Ca2+ sensitivity of the contractile machinery. Passive force measured every 2 min decreased exponentially after the beginning of the trypsin application (t1/2 approximately 12 min). The first 30% decrease of passive force did not affect the stretch-induced variation in Ca2+ sensitivity. Then, with further decrease in passive force, DeltapCa50 decreased. At the lowest passive force investigated 20% of initial passive force, DeltapCa50 decreased by approximately 55%. These effects were not accompanied by a significant modification of either maximal activated force at pCa 4.5 solution or pCa50 at 1.9 microm sarcomere length. This indicates that there was no major functional alteration of the contractile machinery during the protocol as also suggested by contractile and regulatory protein electrophoresis on 2.5-12% gradient and 15% SDS-PAGE gels. Thus, besides modulation induced by the reduced lattice spacing during enhanced heart refilling, Ca2+ sensitivity of the cardiac contractile machinery may be controlled at least partially by internal passive load, which is known to be largely attributable to titin. PMID- 10371697 TI - Repeated cardiac pacing extends the time during which canine hearts are protected against ischaemia-induced arrhythmias: role of nitric oxide. AB - Right ventricular pacing in lightly anaesthetized dogs (4x5 min periods at a pacing rate of 220 beats/min) protects against the consequences of coronary artery occlusion when this is initiated 24 h after the pacing stimulus. The main purpose of the present experiments was to determine whether repeating the pacing stimulus, at a time when protection from the initial stimulus had faded (48 h), prolonged the protection afforded against ischaemia-induced ventricular arrhythmias and other ischaemic changes (epicardial ST-segment mapping; changes in the degree of electrical inhomogeneity in the ischaemic region). Dogs were paced on two occasions, with a 48 h period between and, at different times (48, 72 and 96 h) after the second pacing stimulus, were re-anaesthetized and subjected to occlusion of the left anterior descending coronary artery. There was a marked reduction in the severity of ischaemia-induced arrhythmias 48 and 72 h after the second pacing stimulus (reduction in occlusion-induced and reperfusion induced ventricular fibrillation, e.g. at 72 h 0/11 during occlusion and only 3/11 following reperfusion, compared to 7/21 and 10/21 respectively in the controls P<0.05). The protection had disappeared 96 h following the second pacing stimulus. Changes in ST-segment elevation and in the degree of inhomogeneity largely followed these changes in the severity of ventricular arrhythmias. The results suggest the possibility of maintaining protection against life threatening arrhythmias following coronary occlusion by repeating a preconditioning pacing stimulus. We also demonstrate that this prolonged protection afforded by repeated cardiac pacing is mediated by nitric oxide, since the marked antiarrhythmic effect observed, e.g. 72 h after the second pacing stimulus, was abolished when S-(2-aminoethyl)-isothiourea (AEST), a particularly selective inhibitor of iNOS, had been administered before coronary artery occlusion. PMID- 10371698 TI - Overexpression of endothelium nitric oxide synthase reverses the diminished vasorelaxation in the hindlimb vasculature in ischemic heart failure in vivo. AB - After myocardial infarction (MI), nitric oxide (NO)-mediated vasorelaxation is attenuated in both conduit and resistance arteries. To determine if the attenuated vasorelaxation after MI is due to downregulation of eNOS protein, pharmacological, immunoblotting, and gene transfer of eNOS were performed in rats 3 weeks after MI. Gene transfer was accomplished using a "first-generation" serotype 5, replication-deficient, adenoviral vector (1.2 x 10(9) pfus) containing eNOS cDNA in the hindlimb vasculature for 30 min. Five days after infection, overexpression of eNOS protein was confirmed by immunohistochemical staining and immunoblotting. Recombinant gene expression was localized primarily to the vascular endothelial cells. After MI, eNOS protein level decreased (3.3 +/ 0.9 vs 2.1 +/- 0.8 intensity units/microg protein, n=6, P<0.05); after gene transfer it increased (P<0.05) two-fold to 4.3 +/- 1.2 intensity units/microg protein, n=5. There were no changes in hemodynamics in MI rats transfected with eNOS. Acetylcholine (ACh)-stimulated vasorelaxation was decreased (P<0.05) by 30% after MI and was restored to normal with eNOS transfection. Addition of 100 microm NG-nitro-L-arginine methyl ester (L-NAME) abolished the difference between sham, MI, and MI transfected rats. L-arginine (1 mm) restored the ACh-response in MI-transfected rats toward control, but it did not eliminate the difference between MI and sham rats. We conclude that the attenuated endothelial NO-mediated vasorelaxation in the hindlimb after MI is due to a downregulation of eNOS protein and overexpression of eNOS transgene restores normal endothelial NO mediated vasorelaxation. PMID- 10371699 TI - CD36 deficiency has little influence on the pathophysiology of hypertrophic cardiomyopathy. AB - CD36 is homologous with myocardial long-chain fatty acid (LCFA) binding protein and has been suggested to relate to myocardial fatty acid metabolism. Myocardial scintigraphy with iodine-123 15-(p-iodophenyl)-3-(R, S)-methylpentadecanoic acid (BMIPP) revealed an impairment in LCFA metabolism chiefly in the hypertrophic myocardium in hypertrophic cardiomyopathy (HCM). Recently, the incidence of CD36 deficiency has been reported to be high in HCM patients, and CD36 deficiency was proposed as an etiology of hereditary HCM. However, the pathophysiological effect of CD36 deficiency on HCM has not been fully investigated. We analysed the expression of CD36 antigens on both platelets and monocytes obtained from 82 patients with HCM using two-color flow cytometry. Among the study patients, seven patients (8.5%) demonstrated type II CD36 deficiency, whereas type I CD36 deficiency was not detected. Two of 23 patients (8.7%) with a family history of HCM and five of 59 patients (8.5%) without a family history of HCM showed type II CD36 deficiency respectively. Contrary to the previous report, three of 53 patients with asymmetric septal hypertrophy (ASH) (5.7%) and four of 29 patients without ASH (13.8%) showed CD36 deficiency. Moreover, clinical characteristics, scintigraphic findings, echocardiographic data, and hemodynamic findings disclosed no significant differences between the HCM patients showing normal CD36 expression and those with CD36 deficiency. The incidence of CD36 deficiency in HCM patients is not higher than in the general population. Therefore, CD36 deficiency is not a characteristic factor of HCM and has little influence on the pathyphysiology of HCM. PMID- 10371700 TI - Induction of cardiac beta-adrenergic receptor kinase 1 in rat heart failure caused by coronary ligation. AB - Beta-adrenergic receptor kinase 1 (beta ARK1) participates in the desensitization of beta-adrenergic receptors by uncoupling the signal transduction. The present study was designed to examine whether neurohumoral increase is crucial for the activation of beta ARK1 in heart failure. Four weeks after the ligation of rat coronary artery, LV dP/d t max was reduced, cardiac response to isoproterenol was impaired, and ratio of right ventricular weight to body weight, an index of cardiac hypertrophy, was increased. At the same time, beta ARK1 expression and activity were augmented in the hypertrophied hearts. In addition, plasma norepinephrine content was enhanced in accordance with cardiac hypertrophy, cardiac beta ARK1 expression, LV dP/d t max, and LVEDP. These results of the present study suggest that beta ARK1 is augmented in concert with circulating norepinephrine level and that beta ARK1 may account for, at least in part, the cardiac dysfunction in rat with myocardial infarction. PMID- 10371701 TI - Activation of mitogen-activated protein kinases in in vivo ischemia/reperfused myocardium in rats. AB - In this study, we investigate the in vivo activation of mitogen-activated protein kinases (MAPK) as important signal transduction cascades observed after myocardial ischemia/reperfusion. Myocardial continuous ischemia and ischemia/reperfusion was produced in Wistar rats. The activities of MAPKs in the ischemic and ischemia/reperfused regions were measured using an in-gel kinase assay, an in vitro kinase assay and Western blot analysis. Activator protein-1 (AP-1) DNA binding activity was determined using an electrophoretic mobility shift assay. DNA fragmentation was detected as DNA ladders by agarose gel electrophoresis. The p46JNK and p55JNK activities of continuous ischemia were significantly increased at 30 min (5.9 and 4.2 fold, respectively P<0.05). Coronary reperfusion increased both p42ERK and p44ERK activities at 30 min (3.0 and 2.3 fold P<0.01), and both p46JNK and p55JNK activities at 30 min (1.4 and 1.7 fold P<0.05). The AP-1 DNA binding activities of continuous ischemia were significantly increased at 1, 3 and 7 days (28, 21 and 17 fold, respectively P<0.01). Coronary reperfusion markedly decreased AP-1 DNA binding activities at 1 (41%P<0.01) and 3 days (48%P<0.05). Myocardial DNA fragmentation was considerably more enhanced by reperfusion than continuous ischemia. In conclusion, our present work provides the first in vivo evidence that ERK and JNK are activated by reperfusion from the activities of continuous ischemia. These signal transduction mechanisms may be partially responsible for the myocardial injury. PMID- 10371702 TI - Regulatory mechanisms of calponin phosphorylation in endothelin-1-induced contraction of porcine coronary artery. AB - Calponin is an actin-associated protein that appears to play an auxiliary regulatory role in the contraction of smooth muscle. We report here on the mechanisms for regulation of calponin phosphorylation in the endothelin-1-induced contraction of porcine coronary artery. Treatment of strips of porcine artery with endothelin-1 increased calponin phosphorylation and contraction in a concentration-dependent manner. The time course of the phosphorylation was biphasic, with the response to endothelin-1. The extent of phosphorylation reached a maximum within 5 min of stimulation with 10(-7)M endothelin-1 and then it declined rapidly to reach a minimum at 20 min. A potent inhibitor of protein kinase C, GF109203X, inhibited both calponin phosphorylation and contraction that were induced by endothelin-1 at 5 min, without an inhibition for myosin light chain phosphorylation. Protein phosphatase inhibitor, okadaic acid, had no effect on the extent of phosphorylation at 5 min, but it significantly inhibited the subsequent decrease in calponin phosphorylation. In contrast, in PDBu-treated strips of coronary artery, okadaic acid caused a significant steady increase of the extent of calponin phosphorylation. Our results suggest that calponin phosphorylation might be regulated by protein kinase C and okadaic acid sensitive protein phosphatases, in the endothelin-1-induced contraction of porcine coronary artery. PMID- 10371703 TI - Cardioprotective effects of grape seed proanthocyanidin against ischemic reperfusion injury. AB - There is increasing evidence to indicate cardioprotective effects of red wine consumption. Such cardioprotective properties of wine have been attributed to certain polyphenolic constituents of grapes. The purpose of this investigation was to examine whether proanthocyanidins derived from grape seeds possess cardioprotective properties. Rats were randomly divided into two groups: grape seed proanthocyanidin was administered orally to one group of rats (100 mg/kg/day) for 3 weeks while the other group served as control. After 3 weeks, rats were killed, hearts excised, mounted on the perfusion apparatus and perfused with Krebs-Henseleit bicarbonate (KHB) buffer. After stabilization hearts were perfused in the working mode for baseline measurements of contractile functions. Hearts were then subjected to 30 min of global ischemia followed by 2 h of reperfusion. Coronary perfusates were collected to monitor malonaldehyde formation, a presumptive marker for oxidative stress development. At the end of each experiment, the heart was processed for infarct size determination. Peroxyl radical scavenging activity of proanthocyanidin was determined by examining its ability to remove peroxyl radical generated by 2,2'-azobis (2-amidinopropane) dihydrochloride while hydroxyl radical scavenging activity was tested with its ability to reduce 7-OH.-coumarin-3-carboxylic acid. The results of our study demonstrated that proanthocyanidin-fed animals were resistant to myocardial ischemia reperfusion injury as evidenced by improved recovery of post-ischemic contractile functions. The proanthocyanidin-fed group revealed reduced extent of myocardial infarction compared to the control group. Fluorimetric study demonstrated the antioxidant property of proanthocyanidin as judged by its ability to directly scavenge peroxyl radicals. Taken together, the results of this study showed that grape seed-proanthocyanidins possess a cardioprotective effect against ischemia reperfusion injury. Such cardioprotective property, at least in part, may be attributed to its ability to directly scavenge peroxyl and hydroxyl radicals and to reduce oxidative stress developed during ischemia and reperfusion. PMID- 10371705 TI - Cyclooxygenase inhibition converts the effect of nitric oxide synthase inhibition from infarct size reduction to expansion in isolated rabbit hearts. AB - Nitric oxide (NO) and prostacyclin (PGI2) are putative cardioprotective agents. Evidence indicates that there may be a reciprocal relationship involved in the synthesis of NO and PGI2, so that inhibiting the release of one mediator may promote the synthesis of the other. Therefore, we investigated the effects of concomitantly inhibiting NO and PGI2 synthesis, using NG-nitro-L-arginine (L NOARG) or indomethacin, respectively, on infarct size. Langendorff-perfused rabbit hearts were assigned randomly to one of five treatment groups of n=6: control L-NOARG 100 micromol/l; indomethacin 3 micromol/l L-NOARG 100 micromol/l + indomethacin 3 micromol/l; or L-NOARG 100 micromol/l + L-arginine 1 mmol/l. After 30 min regional ischaemia and 120 min reperfusion, infarct size was assessed by tetrazolium staining. Infarct size was reduced significantly in hearts treated with L-NOARG (20.8+/-1.3%) compared to control hearts (34.7+/ 0.4%). This reduction in infarct size was abolished by co-perfusing with a 10 fold excess of L-arginine (30.7+/-1.7%). While indomethacin alone had no effect (33.4+/-2.3%), perfusion with both L-NOARG and indomethacin resulted in a significant increase in infarct size (44.0+/-1.9%) compared to controls. Treatment with L-NOARG alone increased 6-keto PGF1alpha in coronary effluent prior to ischaemia (30.5+/-1.2 vs 16.6+/-1.3 pg/min/g in controls, P<0.05). This effect was reversed by co-perfusion with either L-arginine or indomethacin. These results indicate that the reduction in infarct size by L-NOARG may be due to increased PGI2 release. Concomitant administration of indomethacin negated this effect and revealed an adverse effect of NO synthase inhibition on infarct size. PMID- 10371704 TI - Expression of cardiac calcium regulatory proteins in atrium v ventricle in different species. AB - The duration of contraction in isolated electrically driven preparations from atrium and ventricle of mouse, rat, rabbit, guinea-pig and dog was consistently shorter in atrial compared to ventricular preparations. Overexpression of phospholamban (PLB) in transgenic mice prolonged duration of contraction, underscoring the importance of PLB for kinetics of cardiac contractility. The expression of regulatory proteins was studied by Western and Northern blot analysis. In rat myocardium, expression of the sarcoplasmic reticulum Ca2+ ATPase (SERCA) was higher in atrium than in ventricle, as was also observed in the rabbit, guinea-pig and wild-type mouse samples. Canine myocardium, however, had similar levels of SERCA (protein and mRNA) in atrium and ventricle. PLB and calsequestrin on protein and RNA levels were lower in atrium than in ventricle from rat, rabbit, guinea-pig and wild-type mouse. PLB protein and RNA levels were higher in ventricle than in atrium at ages 1 and 5 days postnatally and in adult rats. SERCA protein and RNA levels were higher in ventricle than in atrium at days 1 and 5 after birth, but lower in ventricle than in atrium in adult rats. In dog, the calsequestrin level was identical in atrium and ventricle (protein and mRNA) and PLB did not differ between atrium vs ventricle at the protein level but was lower at the mRNA level. Also, Ca2+ uptake was higher in atrium than in ventricle in the dog samples. The expression of the inhibitory subunit of troponin was unchanged between atrium and ventricle in all species studied (protein and mRNA). In dog, protein expression of triadin and junctin was lower in atrium vs ventricle. Triadin mRNA was not altered in dog atrium vs ventricle. In summary, while the hastened relaxation of atrium vs ventricle correlates in part with the lower expression of PLB and higher expression of SERCA, altered regional expression of other SR proteins handling Ca2+ may also play an important role in some species. PMID- 10371706 TI - Influence of RecA on in vivo virulence and Shiga toxin 2 production in Escherichia coli pathogens. AB - The enterohemorrhagic Escherichia coli (EHEC) O157:H7 strains 933 and 86-24 as well as the uropathogenic E. coli (UPEC) strain 536 were compared with their isogenic rec A mutants and rec A trans -complemented strains in intravenous lethality and lung toxicity assays in mice. While the wild-type EHEC strains were fully virulent, the virulence of the rec A mutants was strongly reduced. Complementation of the EHEC rec A mutants with the cloned E. coli recA gene restored their virulence capacity. The stx2EHEC mutant TUV86-2 as well as its isogenic rec A mutant were completely avirulent in both assays. In contrast, RecA had no influence on the virulence of UPEC strain 536. We conclude that the lethality observed with EHEC is presumably mainly due to Shiga toxin, which is severely down-regulated in the rec A mutants as a result of lacking spontaneous phage induction. Therefore, the EHEC rec A+strains 933 and 86-24 were compared for their Shiga toxin 2 (Stx2) production with the respective rec A-counterparts. The rec A mutants of the EHEC strains were significantly reduced in toxin synthesis and were devoid of Stx2 specific phage production. Complementation of the EHEC rec A mutants with the cloned rec A gene enabled the rec A mutants to restore toxin and phage production. These results suggest that the higher level of Stx2 synthesis in the EHEC strains is the result of a higher level of spontaneous Stx2 specific phage induction, which is controlled by RecA. PMID- 10371707 TI - Complement resistance in Borrelia burgdorferi strain 297: outer membrane proteins prevent MAC formation at lysis susceptible sites. AB - Two variants of Borrelia burgdorferi strain 297, complement-resistant wild-type (WT297) and complement-sensitive mutant (MUT297), were used as a model to study the mechanism of resistance to the alternative complement pathway in this organism. No difference in the quantity of membrane attack complex (MAC) deposition on WT297 and MUT297 was observed after 2 h incubation with normal human serum (NHS), at which time 4% of WT297 and 95% of MUT297 were killed. The polymerization of C9 bound to WT297 and MUT297 was demonstrated by immunoblotting using an anti-C9 polyclonal antibody. Immunofluorescence and thin-section immunoelectron microscopy showed MAC to be diffusely distributed on the outer membrane of both variants. Furthermore, MAC appeared to be tightly bound to the surface of both variants as demonstrated by elution studies. Protease treatment rendered WT297 susceptible to killing by NHS, suggesting that outer membrane proteins may be associated with complement resistance of WT297. One- and two dimensional gel electrophoreses showed that proteins of 20 and 30 kDa, and 66 kDa were present in WT297 but were absent or sparse in trypsin-treated WT297 and MUT297. Interestingly, immunoblotting using a polyclonal antibody against C3 showed that C3 fragments appeared to bind different acceptors on WT297 than on trypsin-treated WT297, or MUT297. Therefore, the binding of C3 fragments to acceptors on WT297, in contrast to MUT297, may not direct the formation of the MAC to lysis-susceptible sites on the surface of the bacterium, resulting in the complement resistance of WT297. PMID- 10371708 TI - Differential induction of NO synthesis by gram-positive and gram-negative bacteria and their components in bovine monocyte-derived macrophages. AB - The ability of Gram-positive and Gram-negative bacteria to promote the induction of NO synthesis in bovine monocyte-derived macrophages (MDM) was tested. Heat killed Gram-negative organisms induced NO synthesis at low concentrations (optimum 0.2 to 2 microg/ml wet weight), regardless of the strain, and the response was only moderately enhanced by co-administration of recombinant bovine interferon-gamma (rboIFN-gamma). The activity was largely, but not exclusively, due to lipopolysaccharide (LPS), since it was largely abrogated by co-incubation with polymyxin-B. Diphosphoryl-lipid-A and rough-strain LPS were two orders of magnitude more active than monophosphoryl-lipid A, but two orders of magnitude less active than smooth-strain LPS, suggesting that O side chains contribute to increasing the affinity of LPS or to act as a costimulus. Gram-positive bacteria as single stimuli were four orders of magnitude less potent in inducing NO synthesis than Gram-negative organisms, but upon costimulation with rboIFN-gamma, some of them were excellent inducers of NO synthesis. A similar rboIFN-gamma enhanced NO synthesis induction was also observed for zymosan, muramyl dipeptide, lipoteichoic acid and lipoarabinomannan, although to a lesser extent than for the whole heat-inactivated prototypic organisms. Thus, bovine macrophages exposed to rboIFN-gamma have mechanisms by which they universally react to bacterial compounds distinct from LPS by induction of NO synthesis. PMID- 10371709 TI - Immunization of mice with live oral vaccine based on a Salmonella enterica (sv Typhimurium) aroA strain expressing the Escherichia coli O111 O antigen. AB - Shiga toxin-producing Escherichia coli strains of serogroup O111 are the most frequently isolated non-O157 strains causing outbreaks of gastroenteritis with haemolytic uraemic syndrome (HUS). O antigen is a major antigen in Gram-negative bacteria, and it has been shown that O111 is a protective antigen. Attenuated Salmonella enterica sv Typhimurium aroA strain STM-1 was used as a live carrier to express the E. coli O111 O antigen. Mice immunized intraperitoneally produced serum immunoglobulin G, and mice immunized orally produced serum immunoglobulin G and secretory immunoglobulin A in the intestine against E. coli O111 cells as determined by enzyme-linked immunosorbent assays. PMID- 10371711 TI - Clenbuterol administration into the basolateral amygdala post-training enhances retention in an inhibitory avoidance task. AB - The present study examined the effects of post-training infusions of the specific beta2-adrenergic agonist clenbuterol into the basolateral nucleus of the amygdala (BLA), on inhibitory avoidance retention. Male Sprague-Dawley rats were surgically implanted with cannulae aimed at the BLA. Animals were microinfused with different doses of clenbuterol (from 1.0 to 1000.0 ng) immediately after training, and retention was tested 48 h later. Clenbuterol dose dependently affected retention performances. Whereas the 10.0-ng dose of clenbuterol induced a significant increase in the performances, the groups injected with the other doses (1.0, 100.0, and 1000.0 ng) of clenbuterol did not differ from the vehicle injected animals. These findings, indicating that clenbuterol enhanced retention of the inhibitory avoidance task, are in agreement with previous evidence showing that the adrenergic system of the amygdala is involved in the modulation of memory. Moreover, our data support the hypothesis that the memory-modulating effect of the amygdala adrenergic system is mediated, at least in part, by the activation of the beta-adrenoceptors in the BLA. These findings provide further evidence that the BLA is an important brain region in integrating hormonal influences on memory storage. PMID- 10371710 TI - Microinfusions of flumazenil into the basolateral but not the central nucleus of the amygdala enhance memory consolidation in rats. AB - Extensive evidence indicates that benzodiazepine receptors in the amygdala are involved in regulating memory consolidation. Recent findings indicate that many other drugs and hormones influence memory through selective activation of the basolateral amygdala nucleus (BLA). This experiment examined whether the memory modulatory effect of flumazenil, a benzodiazepine receptor antagonist, selectively involves the BLA. Bilateral microinfusions of flumazenil (12 nmol in 0.2 microl) into the BLA of rats administered immediately after training in an inhibitory avoidance task significantly enhanced 48-h retention performance whereas infusions into the central nucleus were ineffective. These findings indicate that the BLA is selectively involved in mediating flumazenil's influence on memory storage and are thus consistent with extensive evidence indicating that the BLA is involved in regulating memory consolidation. PMID- 10371712 TI - Inferior olive lesions impair concurrent taste aversion learning in rats. AB - Taste aversion learning can be established according to two different procedures, concurrent and sequential. For the concurrent task, two different taste stimuli are offered at the same time, one associated with simultaneous intragastric administration of an aversive stimulus and the other associated with physiological saline. This discrimination is learned by sham-lesioned control animals and by animals with lesions in the cerebellar cortex but not by rats lesioned in the inferior olive. At the same time, animals with lesions in the inferior olive and sham-lesioned animals achieve sequential learning when the gustatory stimuli are offered individually during each daily session. The results obtained show that electrolytic lesions in the inferior olive impair acquisition of concurrent learning and are analyzed in terms of an anatomical system consisting of the vagus nerve, inferior olive, and cerebellum, which differentiates between the two modalities of taste aversion learning, concurrent and sequential. PMID- 10371713 TI - Sequential training in separate paradigms impairs second task consolidation and learning-associated modulations of hippocampal NCAM polysialylation. AB - As transient and time-dependent modulations of neural cell adhesion molecule polysialylation (NCAM PSA) are associated with morphofunctional change and required for the consolidation of spatial and nonspatial forms of learning, we determined the demands imposed on this system by sequential training in the Morris water maze followed by the passive avoidance paradigm. Animals trained in this manner had recall of the water maze but not the passive avoidance response as judged by their escape and avoidance latencies, respectively. Activation of NCAM PSA on dentate neurons at the 12-h post-training time suggested information processing; however, this was significantly less than that predicted for coincident acquisition of both tasks. When sequential training was separated by an interparadigm period of 2 h, an enduring NCAM PSA activation was observed which was indistinguishable from the sum of the expected activations for each individual task. These observations suggest that the NCAM PSA response may become saturated when alternate tasks are presented without an intervening period. PMID- 10371714 TI - Support for a bimodal role for type II adrenal steroid receptors in spatial memory. AB - We investigated the effects of acute adrenal steroid treatment on spatial memory using the Y-maze and employing adrenal steroid receptor antagonists and agonists. For receptor activation, adrenalectomized rats were injected 2 h prior to their first Y-maze trial with sesame oil (adrenalectomy or SHAM), stress levels of corticosterone, a Type I receptor agonist (aldosterone), or a Type II receptor agonist (RU362). For receptor inactivation, unoperated rats were injected with a Type I receptor antagonist (RU318), a Type II receptor antagonist (RU555), sesame oil, or not injected at all. The findings indicated that spatial memory was impaired when the Type II receptors were blocked (RU555) or highly occupied (corticosterone or RU362) and normal for the other treatment conditions. These data suggest that the Type II receptors may be responsible for the inverted U shaped relationship between spatial memory and corticosterone levels reported by others. PMID- 10371715 TI - Rule-based learning impairment in rats with lesions to the dorsal striatum. AB - The present study examined the effects of lesions to the dorsal striatum (DS) in Sprague-Dawley rats, when tested on the acquisition and successive shifts in the position of a goal arm in an eight-arm radial maze. In the procedure we used, rats had to retrieve the location of one baited arm among the eight arms of the maze after it had just been presented as a sample during a forced trial. After attainment of a fixed learning criterion, rats were submitted to five successive shifts in the goal location. Results showed that DS rats were able to learn the position of the goal arm during the acquisition phase as efficiently as sham operated rats. In contrast, when the position of the goal arm was shifted, although DS rats were able to learn its new position, they made significantly more errors and required more sessions to reach criterion than sham-operated rats. These results suggested that both groups did not solve the task using the same behavioral strategy. The analysis of responses made suggested that sham operated rats solved the task using the pairing rule between the forced and the free run (matching-to-sample rule), while DS rats solved the task using only visuo-spatial processing. These data therefore suggest that the dorsal striatum plays an important role in rule-learning ability. PMID- 10371716 TI - Post-training glucose administration attenuates forgetting of passive-avoidance conditioning in 18-day-old rats. AB - The study of memory modulation in infant rats has typically focused on reminder/retrieval treatments involving reexposure to components of the internal or external training context. Rarely have studies employed pharmacological treatments to investigate the neurochemical substrates of memory storage in preweanling rats. The present study investigated the effect of 100 mg/kg of glucose, a common memory modulator in adult mammals, on memory for passive avoidance conditioning in 18-day-old Sprague-Dawley rats. Subjects that were administered an immediate post-training injection of glucose performed significantly better, on a retention test 24 h following training, than those animals that received saline. The glucose group also performed comparably to a control group that was tested 10 min following training. These results are consistent with those of the memory modulation literature in adults and suggest that the rapid rate of forgetting in immature organisms may be the result of a deficiency in a general memory modulatory system. PMID- 10371717 TI - Microtubule organization in germinated pollen of the conifer Picea abies (Norway spruce, Pinaceae). AB - The organization of microtubules in germinated pollen of the conifer Picea abies (Norway spruce, Pinaceae) was examined using primarily confocal microscopy. Pollination in conifers differs from angiosperms in the number of mitotic divisions between the microspore and the sperm and in the growth rate of the pollen tube. These differences may be orchestrated by the cytoskeleton, and this study finds that there are important functional differences in microtubule organization within conifer pollen compared to the angiosperm model systems. Pollen from P. abies contains two degenerated prothallial cells, a body cell, a stalk cell, and a vegetative cell. The body cell produces the sperm. In the vegetative cell, microtubules form a continuous network from within the pollen grain, out through the aperture, and down the length of the tube to the elongating tip. Within the grain, this network extends from the pollen grain wall to the body and stalk cell complex. Microtubules within the body and stalk cells form a densely packed array that enmeshes amyloplasts and the nucleus. Microtubule bundles can be traced between the body and stalk cells from the cytoplasm of the body cell to the adjoining cell wall and into the cytoplasm of the stalk cell. Body and stalk cells are connected by plasmodesmata. The organization of microtubules and the presence of plasmodesmata suggest that microtubules form a path for intercellular communication by projecting from the cytoplasm to interconnecting plasmodesmata. Microtubules in the elongating tube form a net axial array that ensheathes the vegetative nucleus. Microtubules are enriched at the elongating tip, where they form an array beneath the plasma membrane that is perpendicular to the direction of tube growth. This enriched region extends back 20 MUm from the tip. There is an abrupt transition from a net perpendicular to a net axial organization at the edge of the enriched region. In medial sections, microtubules are present in the core of the elongating tip. The organization of microtubules in the tip differs from that seen in angiosperm pollen tubes. PMID- 10371718 TI - Biomechanics of the columnar cactus Pachycereus pringlei. AB - We report the longitudinal variations in stiffness and bulk density of tissue samples drawn from along the length of two Pachycereus pringlei plants measuring 3.69 and 5.9 m in height to determine how different tissues contribute to the mechanical stability of these massive vertical organs. Each of the two stems was cut into segments of uniform length and subsequently dissected to obtain and mechanically test portions of xylem strands, stem ribs, and a limited number of pith and cortex samples. In each case, morphometric measurements were taken to determine the geometric contribution each tissue likely made to the ability of whole stems to resist bending forces. The stiffness of each xylem strand increased basipetally toward the base of each plant where stiffness sharply decreased, reaching a magnitude comparable to that of strands 1 m beneath the stem apex. The xylem was anisotropic in behavior, i.e., its stiffness measured in the radial and in the tangential directions differed significantly. Despite the abrupt decrease in xylem strand stiffness at the stem base, the contribution made by this tissue to resist bending forces increased exponentially from the tip to the base of each plant due to the accumulation of wood. A basipetal increase in the stiffness of the pith (and, to limited extent, that of the cortex) was also observed. In contrast, the stiffness of stem rib tissues varied little as a function of stem length. These tissues were stiffer than the xylem in the corresponding portions of the stem along the upper two-fifths of the length of either plant. Tissue stiffness and bulk density were not significantly correlated within or across tissue types. However, a weak inverse relationship was observed for these properties in the case of the xylem and stem rib tissues. We present a simple formula that predicts when stem ribs rather than the xylem strands serve as the principal stiffening agents in stems. This formula successfully predicted the observed aspect ratio of the stem ribs (the average quotient of the radial and tangential dimensions of rib transections), and thus provided circumstantial evidence that the ribs are important for mechanical stability for the distal and younger regions of the stems examined. PMID- 10371719 TI - Increased nuclear DNA content in developing cotton fiber cells. AB - The nuclear DNA content of developing cotton fiber cells (Gossypium hirsutum, cv. MD51ne) increases ~24% after 2 d postanthesis (dpa). The amount of nuclear DNA at 2 dpa is 5.4 +/- 0.27 pg. At 3-4 dpa it increases to 6.7 +/- 0.24 pg and by 5 dpa it is 6.8 +/- 0.70 pg. These values were obtained by nuclear fluorescence after staining with Hoechst 33258. Human oral squamous cell nuclei were used as a DNA standard. Nuclear DNA content increases in fibers growing on either fertilized or unfertilized ovules. The increase also is detectable in Feulgen stained nuclei using two-wavelength cytospectrophotometry. All measurements were made on isolated fiber cell nuclei using a newly developed method tailored to cotton fiber cells. The results imply that during the early stages of development fiber cell nuclei either selectively amplify certain sequences or enter S-phase replicating a portion of their genome. PMID- 10371720 TI - The morphology of Cercophora palmicola (Lasiosphaeriaceae). AB - A detailed study of ascomal morphology and development in Cercophora palmicola showed that ontogeny is ascohymeniaceous, giving rise to an ostiolate perithecium. Ascomal initials consist of a coiled ascogonium surrounded by several layers of hyphae whose cells become pseudoparenchymatous. The centrum of the young ascoma is composed of a few rows of large, thin-walled pseudoparenchymatous cells that line the ascomal wall, with the central region filled by tightly packed, filamentous paraphyses. The ascogenous system forms along the inside of the layer of pseudoparenchymatous cells at the base of the paraphyses and gives rise to unitunicate asci that grow up among the paraphyses. The wall of the mature perithecium is greatly thickened. It is composed of three regions: a thin outer region of darkly pigmented, angular cells with thickened walls; a broad central region of cells with gelatinized walls; and a thin inner region of flattened cells. Ascomal ontogeny in C. palmicola conforms well to the Sordaria type of development, as defined by Huang. PMID- 10371721 TI - Salinity as a constraint on growth of oligohaline marsh macrophytes. I. Species variation in stress tolerance. AB - The effects of increased salinity on plant growth were examined in a greenhouse experiment with four species common to oligohaline marshes of the northern Gulf of Mexico: Eleocharis palustris, Panicum hemitomon, Sagittaria lancifolia, and Scirpus americanus. Effects of final salinity reached (6 or 12 g/L), salinity influx rate (3 d or 3 wk), and duration of exposure (1, 2, or 3 mo) were investigated. Sagittaria lancifolia was the first species to show visible signs of stress, with browning and curling of older leaf edges. The salt effect was delayed for 6-8 wk in P. hemitomon, but this species had the highest aboveground tissue mortality rate at 12 g/L as exposure continued. Final salt concentration affected all species to a greater degree than did salinity influx rate. No aboveground mortality occurred at 6 g/L, but growth suppression was apparent and varied with species. The magnitude of growth suppression in response to salinity increased for all species as the duration of exposure increased. Overall, we ranked the species as follows, in order from least to most salt tolerant: Panicum hemitomon < Sagittaria lancifolia < Eleocharis palustris < Scirpus americanus. This ranking reflects the field occurrence of these species along a gradient of increasing salinity in northern Gulf of Mexico coastal habitats from freshwater wetlands through oligohaline areas to mesohaline wetlands. PMID- 10371722 TI - Salinity as a constraint on growth of oligohaline marsh macrophytes. II. Salt pulses and recovery potential. AB - The ability of common oligohaline marsh macrophytes of the northern Gulf of Mexico coast to recover from pulses of increased salinity was investigated in a greenhouse experiment with Eleocharis palustris, Panicum hemitomon, Sagittaria lancifolia, and Scirpus americanus monocultures. Components of salinity pulses applied were final salinity reached (6 or 12 g/L), salinity influx rate (3 d or 3 wk), and duration of exposure (1, 2, or 3 mo). After each exposure period, we placed plants into freshwater until the end of the 120-d experiment to determine recovery potential. The four species varied in their ability to recover from the salinity pulses. Within a species, recovery varied with final salinity level and duration of exposure, and to a lesser extent with salinity influx rate. Scirpus americanus, growth of which was stimulated by <3 mo of exposure to 6 g/L, was able to recover even under the most extreme conditions of exposure to 12 g/L salinity for 3 mo. Ability to recover decreased with increased salinity and increased duration of exposure for the remaining three species. Recovery of specific aspects of growth was also suppressed in these species by a rapid salinity influx rate compared to a slow influx rate. The complex variations in recovery patterns displayed by the different species may lead to changes in species dominance following the short-term salinity pulses that can occur during storm events, which in turn may affect marsh plant community composition and structure. PMID- 10371723 TI - Dynamic phenotypic plasticity for root growth in Polygonum: a comparative study. AB - Species differences in patterns of phenotypic plasticity may be an important aspect of adaptive diversity. Plasticity for functionally important root traits was studied in inbred field lineages of Polygonum persicaria and P. cespitosum (Polygonaceae). Replicate seedlings were grown in plexiglass rhizotrons under a range of constant and temporally variable moisture treatments. Plasticity was determined for final whole-plant biomass, root biomass allocation, and absolute and proportional root length. The dynamic aspect of root plasticity was examined by digitizing weekly tracings of the proportional deployment of each plant's root system to different vertical soil layers. Plants of both species expressed significant functionally adaptive phenotypic plasticity in the relative allocation, length, and vertical deployment of root systems in response to contrasting moisture conditions. Plasticity patterns in these closely related species were in general qualitatively similar, but for most traits differed in the magnitude and/or the timing of the plastic response. Dynamic changes in root deployment were more marked as well as faster in P. persicaria. Species differences in patterns of individual plasticity were generally consistent with the broader ecological distribution of P. persicaria in diverse as well as temporally variable moisture habitats. PMID- 10371724 TI - Relative competitive abilities and growth characteristics of a narrowly endemic and a geographically widespread Solidago species (Asteraceae). AB - Relative competitive ability and growth characteristics of the narrow endemic Solidago shortii were compared to those of the geographically widespread S. altissima. Competition and growth studies were conducted over the entire growing season in an ambient-temperature greenhouse, using a 3:1 (v/v) native limestone soil/river sand mixture. Results from a de Wit replacement series experiment (relative yield, relative yield total, plant height, aggressivity values) with S. shortii, S. altissima, and Festuca arundinacea (common competitor) suggested the following competitive hierarchy: S. altissima = F. arundinacea > S. shortii. Using classical growth analysis, we found that the competitive hierarchy was related closely to components of plant size (dry mass, height, leaf area, leaf area duration) and not to relative growth rate or any of its components (net assimilation rate, leaf area ratio, leaf weight ratio, specific leaf area). Solidago shortii allocated proportionately more dry mass to roots (but not to rhizomes) and had significantly greater root/shoot and (root + rhizome)/shoot ratios than did S. altissima. Thus, while the morphological traits of S. shortii enable it to tolerate drier habitats than S. altissima, in moist sites S. shortii easily would be overtopped and shaded out by S. altissima. Low competitive ability may be one of several factors contributing to the narrow endemism of S. shortii. PMID- 10371725 TI - Reproductive biology of Lactoris fernandeziana (Lactoridaceae). AB - Lactoris fernandeziana, monotypic in its family, is endemic to the cloud forests of Robinson Crusoe Island. Although there has been considerable study of the relationships of Lactoris, as a rare species and as a putative primitive paleoherb, little is known of its reproductive biology. Knowledge of the latter is essential for effective conservation programs. The species is gynomonoecious. The overall proportion of flowers is ~1 female:1 hermaphrodite. The inconspicuous semipendulous green flowers, usually in mixed-gender inflorescences, do not produce rewards. Hermaphrodite flowers are herkogamous and protogynous. Pollen grains are shed from the extrorse anthers in permanent dry tetrads. There is a mean of 12879 tetrads per hermaphrodite flower. Both flower types bear an average of ~18 ovules. The P/O (pollen/ovule) ratios imply facultative or obligate xenogamy, but hand pollinations show that Lactoris is self-compatible. No floral visitors were ever observed, but stigmata of open-pollinated flowers bore tetrads, and 64% of such styles had pollen tubes. Flowers enclosed in large mesh (1 mm) bags bore similar numbers of tetrads and pollen tubes. Thus, we conclude that Lactoris is anemophilous, a syndrome perhaps reflected by the P/O ratio. Low genetic diversity (isozymes and DNA) supports selfing and implies limited distance wind pollen dispersal. The small size of the island, the +/- 1000 extant Lactoris plants, coupled with anemophily, self-compatibility, and pendant flower position, have yielded a geitonogamous system with high seed set and low genetic diversity. If inbreeding depression is expressed, it is in seed germination and seedling vigor, for Lactoris is very difficult to cultivate. For this species, effective conservation practices need to focus on habitat preservation and promotion of outcrossing. PMID- 10371726 TI - Further studies of the glandular tissue of the Sauromatum guttatum (Araceae) appendix. AB - Electron microscopic studies showed that the trans-Golgi network (trans indicates the polarity of cisternae within the Golgi apparatus; it is opposite to the cis face that is adjacent to the rough endoplasmic reticulum) was involved in the processing of the osmiophilic material present in the appendix of the inflorescence of Sauromatum guttatum. This material accumulated in the rough endoplasmic reticulum and in special pockets of the plasma membrane prior to heat production. Associations between the endoplasmic reticulum and trans-Golgi network were observed. The Golgi apparatus was composed of 5-6 dictyosomes on one side and one or two somewhat detached cisternae on the other side. Various nonosmiophilic Golgi-derived vesicles were observed: small ones covered with spike-like material, large ones with a smooth surface, and irregularly shaped ones. These electron-translucent vesicles seemed to accumulate in specific localities at the plasma membrane surface in the vicinity of the osmiophilic material; they were not found when the aroma was released. During heat production, the Golgi structures shrank and the activity of the trans-Golgi network seemed to be reduced. At the same time, coated pits were seen at the plasma membrane surface. In some cells, hypertrophic Golgi apparatuses were seen with only 2-3 dictyosomes that contained granulated material in their lumens. Finally, the osmiophilic material was also found in the plasmodesmata. PMID- 10371727 TI - Differential ovule development following self- and cross-pollination: the basis of self-sterility in Narcissus triandrus (Amaryllidaceae). AB - Self-pollination results in significantly lower seed set than cross-pollination in tristylous Narcissus triandrus. We investigated structural and functional aspects of pollen-pistil interactions and ovule-seed development following cross- and self-pollination to assess the timing and mechanism of self-sterility. Ovule development within an ovary was asynchronous at anthesis. There were no significant differences in pollen tube behavior following cross- vs. self pollination during the first 6 d of growth, regardless of style morph type. Double fertilization was significantly higher following cross- vs. self pollination. Aborted embryo development was not detected following either pollination type up to seed maturity. Prior to pollen tube entry, a significantly greater number of ovules ceased to develop following self- vs. cross-pollination. These results indicate that self-sterility in N. triandrus operates prezygotically but does not involve differential pollen tube growth typical of many self-incompatibility (SI) systems. Instead, low seed set following self pollination is caused by a reduction in ovule availability resulting from embryo sac degeneration. We hypothesize that this is due to the absence of a required stimulus for normal ovule development. If this is correct, current concepts of SI may need to be broadened to include a wider range of pollen-pistil interactions. PMID- 10371728 TI - Effects of local density on pollination and reproduction in Delphinium nuttallianum and Aconitum columbianum (Ranunculaceae). AB - Plant populations vary in density both naturally and as a consequence of anthropogenic impacts. Density in turn can influence pollination by animals. For example, plants in dense populations might enjoy more frequent visitation if pollinators forage most efficiently in such populations. We explored effects of plant density on pollination and seed set in the larkspur Delphinium nuttallianum and monkshood Aconitum columbianum. At our site in the Colorado Rocky Mountains, flowers of D. nuttallianum are pollinated primarily by queen bumble bees, solitary bees, and hummingbirds, whereas those of A. columbianum are pollinated primarily by queen and worker bumble bees. We found that the quantity of pollination service to both species (pollinator visitation rate and pollen deposition) was at best weakly related to density. In contrast, seed set declined by approximately one-third in sparse populations relative to nearby dense populations. This decline may stem from the receipt of low-quality pollen, for example, inbred pollen. Alternatively, sparsity may indicate poor environmental conditions that lower seed set for reasons unrelated to pollination. Our results demonstrate the value of simultaneously exploring pollinator behavior, pollen receipt, and seed set in attempting to understand how the population context influences plant reproductive success. PMID- 10371729 TI - Sex ratios and genetic variation in a functionally androdioecious species, Schizopepon bryoniaefolius (Cucurbitaceae). AB - Androdioecy, coexistence of hermaphrodites and males, is an extremely rare breeding system in angiosperms. In the present study, Schizopepon bryoniaefolius (Cucurbitaceae) was confirmed to be functionally androdioecious based on observations of floral and pollen morphology and bagging experiments. Six out of the 11 studied populations consisted of only hermaphrodites, while the other five populations were androdioecious and the frequencies of males were consistently lower than those of hermaphrodites (5.5-28.3%). To understand the consequences of an androdioecious breeding system, genetic variation was estimated using four polymorphic allozyme loci. The degree of genetic differentiation among 11 populations was high (G(ST) = 0.688). Inbreeding coefficients (F(IS)) for all loci significantly deviated from zero. In particular, the F(IS) values averaged across the polymorphic loci in hermaphrodite populations were close to unity, suggesting that hermaphrodites are predominantly selfing in the absence of males. A significant negative correlation was found between the frequencies of males and inbreeding coefficients, indicating that outcrossing rates depend on the population sex ratio. PMID- 10371730 TI - Molecular phylogenetics of Diseae (Orchidaceae): a contribution from nuclear ribosomal ITS sequences. AB - We present here the first molecular phylogeny of tribe Diseae (Orchidoideae: Orchidaceae). Nuclear ribosomal ITS1, 5.8S rDNA, and ITS2 sequences were compared for 30 Diseae, 20 Orchideae, and four Cranichideae and Diurideae outgroups. ITS - rDNA sequences exhibited a transition:transversion ratio of 1.3 and extensive ITS length polymorphism. Phylogenetic analyses using maximum parsimony identified seven major core orchidoid groups. The branching order of the five Diseae and two Orchideae clades was weakly supported but indicated paraphyly of Diseae, with Disperis sister to the rest, followed by successive divergence of Brownleea, Disinae, Coryciinae sensu stricto (s.s.), Satyriinae, and terminated by Orchidinae plus Habenariinae. Within the monophyletic Disinae, Herschelia and Monadenia were nested within a paraphyletic Disa and clustered with D. sect. Micranthae. Within monophyletic Satyriinae, Satyridium rostratum plus Satyrium bicallosum was sister to the rest of Satyrium, and then Satyrium nepalense plus S. odorum was distinct from a cluster of six species. Coryciinae are paraphyletic because Disperis is sister to all other core orchidoids. Coryciinae s.s. are sister to Satyriinae plus Orchideae, with Pterygodium nested within Corycium. Maximum likelihood analysis supported possible affinities among Disinae, Brownleeinae, and Coryciinae but did not support monophyly of Diseae or an affinity between Disinae and Satyriinae. Morphological characters are fully congruent with the well-supported groups identified in the ITS phylogeny. PMID- 10371731 TI - In situ fossil seedlings of a Metasequoia-like taxodiaceous conifer from Paleocene river floodplain deposits of central Alberta, Canada. AB - Fossil seeds and seedlings of a Metasequoia-like taxodiaceous conifer occur in Paleocene deposits at the Munce's Hill and Gao Mine localities of central Alberta, Canada. Compression/impression specimens are preserved in upright growth positions among seedlings of the cercidiphyllaceous dicot Joffrea speirsii Crane & Stockey. There are a large number of seeds, a few of which were buried while germinating and show a radicle or short primary root. More than 500 Metasequoia like seedlings have been identified that have two linear cotyledons with parallel margins and rounded tips. Three specimens have been found that display three cotyledons. Slightly older seedlings show decussate pairs of leaves attached to the stem distal to the cotyledons. Still older seedlings have axillary branches that show varying sizes and numbers of opposite leaves arranged in a single plane distal to the opposite pairs. These specimens reveal that both Joffrea and this extinct taxodiaceous conifer were early colonizers of North American floodplain communities at the beginning of the Tertiary. PMID- 10371732 TI - [13th Special Group Session of the Association of Pharmaceutical Chemistry and the German Pharmaceutical Society. Freiburg, Germany, 8-10 March 1999. Abstracts]. PMID- 10371733 TI - [Continuing medical education in urology]. PMID- 10371734 TI - [Urology and grammar]. PMID- 10371735 TI - [Medical treatment of benign prostatic hyperplasia. The efficacy of finasteride depends on prostatic volume]. AB - OBJECTIVE: To demonstrate that the efficacy of finasteride in benign prostatic hyperplasia (BPH) depends on the prostate volume. METHODS: 75 patients with BPH were classified according to the prostate volume: 20-40 Gm (group I), 40-80 Gm (group II) and more than 80 Gm (group III). The average duration of treatment for all three groups was 16.2 months. Patients were evaluated for symptom score (I PSS), quality of life, prostate volume by ultrasound, maximum urinary flow rate, residual urine, plasma PSA and testosterone levels. RESULTS: Ultrasound demonstrated a reduction in prostatic volume of 6.8-14.8 Gm. Symptom score was reduced by 9.2-15.5 points and quality of life 2-2.7 points. Maximum urinary flow rate increased 3.4-4.4 ml/sec and residual urine decreased in all three groups. Testosterone values showed no significant changes. PSA values showed a reduction of 1.3-1.7 ng/ml. CONCLUSIONS: The efficacy of finasteride depends on the prostatic volume. Medical treatment is a temporary alternative to surgery due to its cost and because patients are unaccustomed to having to take a tablet daily. PMID- 10371736 TI - [Perioperative transfusion in prostatic surgery]. AB - OBJECTIVE: To analyze the likelihood of perioperative transfusion in prostate surgery. METHODS: The records of patients who underwent prostate surgery at the Hospital Central de Asturias en 1996 were reviewed. The abstracted patient discharge records, encoded according to the ICD-9-CM, were analyzed for gender, age, diagnosis, surgical and non-surgical procedures, including transfusions of blood products. RESULTS: 29 (14.2%) of 204 operated patients required transfusion. By univariate analysis, the likelihood of requiring transfusion was significantly higher in patients with cancer (28.8%), patients undergoing radical prostatectomy (40%), and those undergoing more than one surgical procedure (21.2%). By multivariate logistic regression analysis, the surgical procedure, age and simultaneous urinary bladder surgery were related with the likelihood of requiring transfusion. The adjusted odds ratio for perioperative transfusion was 10.14 times in radical prostatectomy than in transurethral prostatectomy, 2.67 times for patients more than 75 years than in those aged less, and 2.85 times in patients undergoing simultaneous urinary bladder surgery than those who do not. CONCLUSIONS: Due to the wide range of variability of the adjusted likelihood for transfusion in prostate surgery (3.5% to 83.4%), estimation of the individual likelihood for transfusion could be useful for planning surgical transfusions, pretransfusion tests and autologous transfusion in these patients. PMID- 10371738 TI - [Mid-term results of a program for the monitoring of superficial bladder tumors]. AB - OBJECTIVE: To identify the medium-term clinical prognostic factors for recurrence and progression in superficial bladder tumors and to analyze the efficacy and safety of the surveillance program. METHODS: 135 patients with primary superficial bladder tumor (Ta: 57, T1: 76, Tx: 2, grade I: 51, grade II: 70, grade III: 12, unknown grade: 2) were evaluated by cystoscopy/cytology alternately with ultrasound/cytology during the first two years, and by ultrasound and cytology yearly after the third year. The influence of tumor characteristics (stage, grade, size and number) and patient age and sex on tumor recurrence and progression were analyzed. The disease-free interval was expressed through an actuarial curve. The relationships between the annual recurrence rate and disease-free interval, and tumor and patient characteristics were analyzed. The influence of each variable in the presence of the other covariables was determined by multivariative analysis using a logistic regression model. RESULTS: The recurrence rate was 51.3% and the progression rate was 2.75%. The mean recurrence rate was 0.59 and the mean disease-free interval was 16.2 months. Tumor grade, size and number significantly influenced recurrence. Tumor size and number influenced the annual recurrence rate. Similarly, tumor number and size were associated with a shorter disease-free interval. Logistic regression analysis showed a relationship between tumor number and recurrence (relative risk = 2.7), and identified tumor size as a characteristic of a high recurrence rate (relative risk = 3.3). None of the factors could predict a higher risk for tumor progression. CONCLUSIONS: The estimation of the risk factors shows a wide intra and interindividual variability. It might therefore be necessary to establish them for each institution. In the medium-term, surveillance can be considered effective in terms of the recurrence rate and safe in terms of the progression rate. PMID- 10371737 TI - [Quantification of p53 oncoprotein in bladder carcinoma: 5-year experience]. AB - OBJECTIVE: Mutations in the p53 gene are frequently detected in some types of malignant tumors (bladder, prostate, kidney, lung, breast, colon and rectum). This study analyzed the utility of p53 quantitation in transitional cell carcinoma of the bladder and evaluated the results at 5 years. METHODS: A prospective clinical cohort study was conducted on 81 patients. The study comprised two groups: nontumoral bladder tissue specimens from 20 patients (group I) and tissue specimens from 61 patients with bladder carcinoma (group II). In both groups the tissue specimens were obtained between November 1992 and November 1993, including the follow-up period until July 1998. p53 expression was determined by a quantitative method based on immunoluminiscence (LIA-MAT p53). RESULTS: p53 expression was higher in bladder carcinoma than in healthy urothelial tissue; higher values of p53 were found for infiltrating and undifferentiated tumors. The p53 values were higher in patients with tumor recurrence than in those without (NS). The Bonferroni multiple comparisons test showed a higher mortality in patients with p53 > 0.9 than in patients who are alive or have died from other causes (p = 0.000). CONCLUSION: The results show that p53 LIA-MAT is an independent prognostic factor at a cutoff of 0.9 and permits identification of a subgroup of patients at high risk. PMID- 10371739 TI - [Female urinary incontinence: clinical-urodynamic correlation]. AB - OBJECTIVES: To determine the diagnostic utility of the urinary symptoms and physical examination in the urodynamic diagnosis of the type of female urinary incontinence. METHODS: A descriptive longitudinal study was conducted in 100 women undergoing urodynamic evaluation. The patients were asked about the urinary symptoms and were evaluated by physical examination and cystomanometry. RESULTS: A statistically significant correlation was found for age and clinical type of urinary incontinence. A strong trend was observed for clinical type of urinary incontinence and cystocele grade, and the urodynamic type of incontinence. A low sensitivity was found for the clinical type in relation to the urodynamic type of incontinence. CONCLUSIONS: Urinary incontinence clinical data do not permit determination of the urodynamic type of incontinence. PMID- 10371740 TI - [A decade of laparoscopic varicocelectomy: costs and learning stages]. AB - OBJECTIVE: To present the results of a retrospective study on laparoscopic varicocelectomy, with special reference to the changes in the operating time observed throughout the training period. METHODS: We have reviewed the laparoscopic varicocelectomy procedures performed from 1987 to 1997. The operating times were graphically represented and compared, and the modifications and complications observed over the ten-year period were analyzed. RESULTS/CONCLUSIONS: The operating times decreased as the number of procedures increased and its duration further decreased as the interval between operations became shorter. In our case, the average operating time after the training period has been completed is 44 minutes. The cost of laparoscopic varicocelectomy and laparoscopic surgery in general is comparable to that of open surgery if nondisposable material is utilized. PMID- 10371741 TI - [Percutaneous nephropexy in the treatment of renal ptosis]. AB - OBJECTIVE: Surgical fixation of ptotic kidneys has been utilized since the late nineties of the last century and more than 170 techniques have been described in the literature, all of which are by open surgery. The efficacy of a new percutaneous nephropexy procedure is described herein. METHODS: The technique basically consists in the fixation of the organ in its normal position by fibrous ligation created by a nephrostomy (preferably loop nephrostomy) and a nephropyeloureteral catheter inserted percutaneously. Our technique of percutaneous nephropexy was performed in 15 female patients with marked right renal ptosis, a long history of pain, which was complicated by lithiasis in the same kidney in 5 cases, and in whom medical treatment had repeatedly failed. RESULTS: At 6-14 months' follow-up, all of the patients are asymptomatic with negative urological cultures, no calculi, kidney in normal position and ureter corrected. CONCLUSIONS: The results demonstrate that our percutaneous nephropexy technique is an effective alternative treatment for the old and unresolved problem of renal ptosis. PMID- 10371743 TI - [Neurophysiologic techniques in the diagnosis of erectile dysfunction: study of 105 cases]. AB - OBJECTIVE: To present the results of the neurophysiological diagnostic evaluation in patients with erectile dysfunction. METHODS: 105 consecutive patients with complaints of erectile dysfunction were evaluated. The patients were divided into five groups according to their history: surgery/trauma, diabetes, vascular disease, toxicity, and no antecedents. Neurophysiological examination, laboratory tests, hormone and vascular studies were performed. Patient evaluation included neurophysiological studies on nerve conduction velocity (NCV), bulbocavernosus reflex (BCR), spinal and scalp somatosensory evoked potentials from the posterior tibial nerves (SEPt) and from the dorsal penile nerve (SEPp). In selected cases, electromyography and neuroimaging studies were done. RESULTS: 71 patients (67.62%) showed organic alterations; 57 of these patients had abnormal neurophysiological tests. Ranked in order of incidence, the tests were abnormal for NCV in 46/105 patients, SEPt 37/105, BCR 35/105 and SEPp in 30/105 patients. The incidence of BCR abnormalities was higher than that of the NCV only in the surgery/trauma group, probably due to a higher prevalence of local injuries, while NCV showed the highest incidence of abnormal results in the other groups of patients. Only 8 patients showed normal NCV with abnormal BCR, SEPp and/or SEPt (14.04% of patients had neurophysiological disturbances). CONCLUSION: In this study, most of our patients showed vascular, neurologic or both abnormalities, and were rarely associated with other factors. In our view, NCV is the technique of choice in the assessment of neurological disorders in patients with erectile dysfunction, whereas the other less sensitive tests are considered to be indicated basically in erectile dysfunction highly suspected as having an underlying neurogenic cause. PMID- 10371742 TI - [The impact of creating andrology units on the treatment of erectile dysfunction with intracavernosal injections]. AB - OBJECTIVE: The impact of the creation of the andrology unit on intracavernous injection therapy for erectile dysfunction is analyzed in the present study. METHODS: The records of patients treated by intracavernosal injection of vasoactive drugs for erectile dysfunction from 1990 to 1997 were reviewed. RESULTS/CONCLUSIONS: Demands for solutions to the problem of erectile dysfunction have significantly increased over the last decades. The efficacy of intracavernosal injection of vasoactive drugs in the treatment of erectile dysfunction has been demonstrated. After the creation of the andrology unit within the urology services, patient acceptance of treatment and compliance increased, and complications have diminished. PMID- 10371744 TI - [Rupture of kidney pelvis in retroperitoneal fibrosis]. AB - OBJECTIVE: To present a case of rupture of the renal pelvis in a patient with idiopathic retroperitoneal fibrosis and no previous dilatation of the left excretory system. METHODS/RESULTS: A patient with retroperitoneal fibrosis underwent surgery for ureterolysis and intraperitonealization of the right ureter. The control IVP was unremarkable. One week later, however, rupture of the left renal pelvis was observed, which was resolved by placement of a ureteral catheter. CONCLUSIONS: In the case described, rupture of the renal pelvis without previous obstruction could be due to ureteral akinesia associated with the retroperitoneal fibrosis. PMID- 10371745 TI - [Polypoid eosinophilic cystitis mimicking bladder carcinoma]. AB - OBJECTIVE: To report an additional case of polypoid eosinophilic cystitis mimicking a urothelial carcinoma of the urinary bladder. METHODS: An 81-year-old man presented with hematuria and dysuria that he had noted for the past month. The US findings were consistent with a bladder tumor and consequently the patient was submitted to transurethral resection. RESULTS: Histological analysis disclosed inflammatory infiltrates with numerous eosinophils (eosinophilic cystitis), but no evidence of tumor. The patient is well two years later. CONCLUSIONS: The present case demonstrates that polypoid eosinophilic cystitis can mimick a bladder carcinoma. Histopathological analysis is necessary for the differential diagnosis. PMID- 10371746 TI - [Spontaneous retroperitoneal hemorrhage]. AB - OBJECTIVE: The etiology of spontaneous retroperitoneal hemorrhage is analyzed and the clinical, diagnostic and therapeutic aspects are discussed. METHODS/RESULTS: Two cases of spontaneous retroperitoneal hemorrhage from a ruptured adenocarcinoma of the kidney are reported. Both patients presented the characteristic Lenk's triad. Definitive diagnosis was made on the CT findings and treatment was by nephrectomy. CONCLUSIONS: The most common cause of retroperitoneal hemorrhage arising from the kidney is a tumor. Treatment is based on the underlying cause of the underlying cause of the hemorrhage and the patient's hemodynamic status. PMID- 10371747 TI - [Traumatic unilateral testicular dislocation]. AB - OBJECTIVE: To report a case of traumatic testicular dislocation, an unusual complication of pelvic trauma. METHODS/RESULTS: A case of traumatic testicular dislocation that had been diagnosed two months after the initial injury is presented. CT and color doppler US revealed a viable right testis displaced in the inguinal subcutaneous region. The testis was repositioned in the scrotum and orchidopexy was performed. Follow-up control evaluation at 6 months revealed a normal testis in the right hemiscrotum. CONCLUSIONS: Traumatic testicular dislocation is a rare complication that may be undetected at the time of the initial injury. Surgical reduction and repositioning achieve good results. PMID- 10371749 TI - [Castleman's disease: isolated retroperitoneal mass. Report of a case]. AB - OBJECTIVE: To describe a rare case of vascular hyaline variant of Castleman's disease presenting as a solitary mass, with special reference to the radiological findings and differential diagnosis. The literature is briefly reviewed. METHODS/RESULTS: A 48-year-old male presented with nonspecific pain in the left flank. Routine analyses were unremarkable. Sonographic and CT studies showed a well-defined, highly vascularized, 3.3 x 3.6 x 4 cm retroperitoneal mass. The risk of hemorrhage made a preoperative biopsy impracticable and the mass was removed surgically. The pathological analysis of the surgical specimen showed a localized retroperitoneal angiofollicular lymphoid hyperplasia (vascular hyaline variant). CONCLUSIONS: Castleman's disease can rarely present as a solitary retroperitoneal mass, which must be distinguished from primary retroperitoneal masses that are usually malignant. Imaging techniques are not conclusive. Definitive diagnosis is based on the postoperative pathological findings. PMID- 10371748 TI - [Tumor excision in renal oncocytoma: report of a new case]. AB - OBJECTIVE: To describe a case of renal oncocytoma that was treated by conservative surgery. METHODS: Herein we describe a 55-year-old patient with a preoperative suspicion of left renal oncytoma based on the findings of punction aspiration biopsy and CT. The patient underwent tumor resection. RESULTS/CONCLUSIONS: Oncocytomas can present typical radiological features. Conservative surgery is a treatment option in carefully selected patients that usually achieves good results. PMID- 10371750 TI - [Arterial priapism. Resolution following embolization with autologous thrombus]. AB - OBJECTIVE: To report our experience with the management of arterial priapism by embolization. METHODS: Two patients with arterial priapism secondary to perineal trauma are described. Diagnostic evaluation included intracorporal blood gas, echo Doppler and selective pudendal arteriographic assessment. Treatment was by selective autologous embolization. RESULTS: Detumescence was achieved and control follow-up evaluation at 12 and 24 months demonstrated normal erectile function. CONCLUSIONS: In our view, selective embolization is an effective treatment for arterial priapism arising from perineal trauma. PMID- 10371751 TI - Congenital mesoblastic nephroma (CMN) with an unusual immunohistochemical feature. AB - OBJECTIVES: To describe a case of congenital mesoblastic nephroma (CMN) treated by radical nephrectomy with no evidence of relapses after five years in spite of an unusual positivity for proliferating cellular nuclear antigen (PCNA). METHODS: A three-month-old child presented a right renal mass with compression of the inferior vena cava. Excretory urography showed an intrarenal mass with distortion of the calyceal system. There was no evidence of metastasis. Radical nephrectomy was carried out; no adjuvant therapy was given. Histological and immunohistochemical studies were performed. RESULTS: The tumor was a 6 x 6 x 5 cm solitary mass extending into perirenal tissue, involving the hilar vessels but not the ureter. Histologically, it has been classified as a congenital mesoblastic nephroma of the classic variant. Positive reaction for vimentin and actin was observed. Strong positivity for PCNA and negativity for P53 were revealed. CONCLUSIONS: PCNA is considered to be a reliable marker of potential malignancy. This, however, contrasts with the biological behavior of our case. Further evaluation is required for correct interpretation of this additional information and to avoid inappropriate aggressive therapy. PMID- 10371752 TI - Urinary lithogen risk test: usefulness in the evaluation of renal lithiasis treatment using crystallization inhibitors (citrate and phytate). AB - OBJECTIVES: This paper presents the results of a test to globally determine the urinary risk factor of calcium stone formation in the evaluation of treatments using crystallization inhibitors, such as citrate and phytate. METHODS: Three groups of active calcium oxalate stone-formers have been selected. The lithogen urinary risk was determined using a specially designed disposable test before any medical treatment. After evaluation group I did not receive any treatment, group II was treated with potassium citrate and group III with a phytate-rich dietary complement. When 15 days had elapsed, the test to evaluate the risk of urinary calcium stone formation was applied again to the three groups. The main lithogenic biochemical parameters of each tested urine were also determined before and after treatment. RESULTS: An important number of calcium oxalate stone formers with high urinary risk factor (positive test) became negative after medical treatment (52% of the citrate-treated patients and 50% of the phytate treated patients), but only 7% of the untreated patients (1 patient) showed a decrease in their urinary risk factor for calcium stones (negative test) after 15 days had elapsed. When the treatment was not effective, in an important number of cases, the urine contained high levels of calcium or showed pH values greater than 6.5. CONCLUSION: From the obtained results it can be concluded that the test is useful to evaluate the efficacy of a given renal lithiasis medical treatment, and also the efficacy of the treatment of calcium oxalate renal lithiasis using crystallization inhibitors, such as citrate and phytate, in an important number of cases. PMID- 10371753 TI - A genetic analysis of the eating and attitudes associated with bulimia nervosa: dealing with the problem of ascertainment in twin studies. AB - Little is known about the etiology of bulimia nervosa and the attitudes associated with it. We have undertaken a study of selected (45 pairs) and unselected (106 pairs) female twins to elucidate the broad causes of individual differences in these behaviours and attitudes. The selected sample was chosen on the basis of at least one of the twin pair having a lifetime incidence of bulimia nervosa. Biometrical model fitting, which corrected for the biased twin correlations of the ascertained group, was used to investigate the genetic and environmental risk factors contributing to the development of bulimia nervosa. The best-fitting model showed that individual variation was best explained by additive genetic influences (62%) and nonshared environmental influences (38%). The proportion of genetic variance affecting individual variation in the ascertained group and the random group was not found to be significantly different. In summary, it is suggested that it may not be necessary to supplement a randomly selected sample with an ascertained sample when investigating the liability to a low-prevalence psychiatric disorder if a continuous measure of that disorder is available. PMID- 10371754 TI - Half of the variation in susceptibility to mortality is genetic: findings from Swedish twin survival data. AB - Molecular epidemiological studies confirm tremendous variability in genetic and environmental susceptibility to disease and death for humans. This variability as well as the roles of genetic and environmental factors in susceptibility to death can be estimated in the analysis of survival data on related individuals (e.g., twins). In this paper, correlated gamma-frailty models are applied to survival data on Swedish twins to estimate genetic parameters in six models of susceptibility. It is shown that the frailty model with additive genetic and nonshared environmental components fits the data best. The estimate of narrow sense heritability in gamma frailty is about 50%. The results of genetic analysis confirm our earlier findings from the studies of Danish twins that about 50% of individual susceptibility approximated by gamma-distributed frailty is heritable. PMID- 10371755 TI - On the relationships of high-frequency hearing loss and cochlear pathology to the acoustic startle response (ASR) and prepulse inhibition of the ASR in the BXD recombinant inbred series. AB - The measurement of the acoustic startle response (ASR) and prepulse inhibition (PPI) of the ASR in many inbred strains of mice, including C57BL/6 and DBA/2, may be complicated by age-related high-frequency hearing loss (HFHL) and the associated cochlear pathology. Willott and Erway (1998) have recently reported on the age-related changes of the acoustic brain response in the BXD recombinant inbred (RI) series. Based on these data, the RI series was divided into three groups: juvenile-, intermediate-, and adult-onset HFHL. Each of these groups was tested using paradigms which varied the frequency or intensity of the auditory startle and prepulse stimuli. The results obtained in adolescent mice (6-8 weeks) demonstrate that ASR performance is independent of HFHL; there was no group dependent decline in the ASR amplitudes for high-frequency stimuli. The expected effect of HFHL on PPI is to increase the salience of the still-audible tones. In response to a white-noise prepulse stimulus, the PPI in the juvenile-onset group (which shows marked HFHL at 6 weeks) was similar to that in the adult-onset group. However, when the prepulse stimulus was a pure tone, the juvenile group showed a decrease in salience across all frequencies tested (5-20 kHz). The data point out the need for carefully constructing auditory tasks in the BXD RI series, to avoid the confounding effects of HFHL. PMID- 10371756 TI - Cytoplasmic factors do not contribute to a maternal effect on ethanol teratogenesis. AB - Both maternal and fetal genetic factors influence variations in response to prenatal ethanol exposure. To assess the effect of maternal genotype on the incidence of ethanol teratogenesis, a reciprocal cross study was conducted in an animal mode using the relatively susceptible C57BL/6J (B6) and the relatively resistant DBA/2J (D2) inbred mice. This mating pattern produced four embryonic genotypes: true-bred B6B6 and D2D2 litters and hybrid B6D2 and D2B6 litters. To examine the role of maternal egg cytoplasm as the source of variation that could account for a maternal effect, B6D2 and D2B6 F1 females were mated back to B6 males, which produced two additional embryonic genotypes: B6D2.B6 and D2B6.B6. Dams were intubated with either 5.8 g/kg of ethanol or an isocaloric amount of maltose-dextrin on day 9 of pregnancy. On day 18 of pregnancy, dams were sacrificed, fetuses were removed, weighed, sexed, and examined for gross morphological malformations. Every other fetus within a litter was prepared for either skeletal or soft tissue analysis. Results showed a higher rate of teratogenesis in the B6D2 group compared to the genetically similar D2B6 group, which indicates an influence of maternal genotype on susceptibility to ethanol teratogenesis. The percentage of affected male and female fetuses did not differ, which suggests that sex-linked factors are not responsible for the maternal effect. The backcross B6D2.B6 and D2B6.B6 litters did not differ significantly for any measure of teratogenesis, suggesting that differences in maternally transmitted cytoplasmic material are not the cause of the maternal effect. Factors that could account for the maternal effect are differences in the maternal uterine environment and genomic imprinting. Separating maternal from fetal-mediated mechanisms responsible for susceptibility to ethanol teratogenesis is needed for identifying mothers and infants at risk. PMID- 10371757 TI - Rearing environmental enrichment in two inbred strains of mice: 1. Effects on emotional reactivity. AB - The effects of an enriched rearing environment on two types of anxiety-like behavior (designated "trait" and "state" anxiety) and on spontaneous activity were investigated in two inbred strains of mice, BALB/c (C) and C57BL/6(B6). Subjects were socially reared from birth to 56 days of age under enriched or standard rearing conditions. The enriched environment consisted of an assembly of plastic boxes in which a various number of objects (running wheels, pieces of plastic, etc.) offered the possibility of multiple activities. The subjects were subsequently tested in three situations: a spontaneous activity recorder, an elevated plus-maze test (a model of state anxiety), and a free exploration test (a model of trait anxiety). No group differences could be found in spontaneous activity. Environmental enrichment, however, decreased the level of both types of anxiety-like behavior in the C strain. In contrast, the level of trait anxiety of the B6 mice was not modified. The results were discussed in relation to possible CNS modifications, especially in the limbic system. PMID- 10371758 TI - Selective breeding for high and low alcohol preference in mice. AB - High and low alcohol preference (HAP and LAP, respectively) mice were created by 10 generations of bidirectional selection for differences in two-bottle choice alcohol consumption. The progenitors used for selection were HS/lbg mice, which are a genetically defined, outbred stock. During selection, mice had 24-h, daily access to 10% alcohol (v/v) and water ad libitum for 30 days and were selected based on the alcohol (g/kg) consumed per day over the entire period. Food was available ad libitum. At S10, line means for alcohol consumption differed greatly, with consumption of over 12 g/kg per day in the HAP mice and less than 2 g/kg per day in the LAP mice. Realized heritability for bidirectional selection was approximately 0.2. Female mice consumed more alcohol than male mice. There were no differences between lines in alcohol elimination rate, nor were there line differences in intake of salt or quinine solutions. However, consumption of saccharin solutions was greater in HAP mice than LAP mice, consistent with previous findings of a genetic correlation between sweet preference and alcohol drinking. Because the mouse genome is relatively well characterized, these selected lines should prove a useful tool for assessment of the genetic basis of, and phenotypes that correlate with, alcohol drinking. PMID- 10371759 TI - Comparison of two sensitization paradigms of the acoustic startle response in Wistar and Sprague-Dawley rats. AB - An increase in general responsiveness after aversive stimulation has provided a most widely accepted and well-understood sensitization paradigm. According to a second paradigm (based on the dual process theory of habituation and sensitization), not only additional aversive stimuli, but also the response eliciting stimuli themselves, induce sensitization. To relate these two sensitization paradigms, we compared the course of startle response parameters during repetitive acoustic stimulation with the change in startle amplitude after electric footshocks in outbred Wistar and Sprague-Dawley rats. Compared to the Wistar rats used, the Sprague-Dawley rats showed a lower response decrement and a shortened latency during repetitive stimulation, both of which are indicators of increased sensitization by the startle-eliciting stimuli. In addition, the Sprague-Dawley rats also demonstrated a reduced increase in startle amplitude following footshock. This was postulated to be a consequence of the strong sensitization by startle-eliciting stimuli, which interferes with sensitization elicited by footshock. Because our Wistar and Sprague-Dawley rats did not differ in initial startle amplitude, but mainly in susceptibility to sensitization, further comparisons of these genetically different stocks of rats seem to be of potential value in studying differences in fear-motivated behavior. PMID- 10371760 TI - Developmental isolation and subsequent adult behavior of Drosophila paulistorum. V. Survey of six sibling species. AB - In an investigation into the effects of developmental isolation from all conspecifics, the Drosophila willistoni group of six sibling species responded to differing degrees: all six are reproductively isolated from D. paulistorum, the tester species. Drosophila pavlovskiana, a narrow endemic, proved the most vulnerable, responding by reducing its adult sexual isolation, if eggs, any instar, and sometimes even pupae were socially isolated. To lesser degrees, D. tropicalis and D. willistoni both produced similar results only when their eggs were isolated, i.e., when from the moment of egg deposition on, there was absolutely no contact with other flies until testing for mating behavior. The remaining siblings, D. equinoxialis and D. insularis, were immovable. PMID- 10371761 TI - [Chronic renal insufficiency. A permanent public health problem]. AB - Chronic renal insufficiency raises an ever-increasing public-health problem due to its permanent growth among the general population and the escalating cost of renal replacement therapies. By the end of 1995 there were close to 33,700 patients with end-stage renal failure maintained alive with renal replacement methods in France. About 11,200 had a functioning kidney graft, whereas 22,500 were treated with various dialysis techniques, in and out-of-center hemodialysis and peritoneal dialysis. An optimal health policy should contribute both to prevent renal insufficiency and offer each patient his/her best specific mode of treatment at the lowest cost for the community. Renal transplantation should be much more widely promoted and utilized through measures aiming at reducing the too high refusal rates of organ donation in subjects with brain-death. Promotion and extension of out-of-center dialysis techniques are also necessary. Design of reliable epidemiological studies dealing not only with end-stage renal failure patients but with the early stage and time-course of renal insufficiency is also mandatory. A deeper investigation in the area of renal-risk factors and a qualified follow-up of patients with mild/moderate renal insufficiency are essential to avoid or delay an evolution towards end-stage renal failure. Prevention of renal fibrosis has a central role in such a long-term public health policy. PMID- 10371762 TI - [Inflammatory mechanisms of renal fibrosis: glomerulonephritis]. AB - Studies of glomerulonephritis models have shown that inflammatory reaction is responsible for the development of glomerulosclerosis and tubulo-interstitial sclerosis and, hence, for the progression to end stage renal failure. That macrophage accumulation and fibrosis extension are frequently not closely related events suggests that macrophages are not involved in progression process. Glomerular sclerosis is rather associated with the release of mediators from resident cells-mainly growth factors such as platelet-derived growth factor and transforming growth factor-beta--the synthesis and bioactivity of which are enhanced by inflammatory mediators. Tubulo-interstitial sclerosis is induced by inflammatory lesions of the glomerulus that lead to proteinuria. Indeed, reabsorption of proteins in proximal tubule triggers epithelial cells to release proinflammatory and prosclerotic mediators into the interstitium. New therapeutic approaches including gene transfer strategies are directed at suppressing the efficiency of such mediators. PMID- 10371763 TI - [Vascular mechanisms of renal fibrosis. Vasculonephropathies and arterial hypertension]. AB - Nephrovasculopathies are an increasing cause of end-stage renal failure. Nephrosclerosis is a common finding in the hypertensive patient. However, genetic factors play a prominent role in its incidence. Nephrosclerosis is a common cause of early renal failure in blacks of African ancestry, as opposed to white Europeans, in whom hypertensive nephrosclerosis rarely and slowly leads to uremia. That primary hypertension is accompanied by arterionephrosclerosis and arteriolonephrosclerosis, by focal and segmental glomerulosclerosis leading to glomerular obsolescence and by interstitial fibrosis has been established for nearly a century. However, renal vascular lesions can be observed in animal models as well as in some humans, especially blacks, in the absence of, or preceding the onset of hypertension. This suggests that nephroangiosclerosis might stem from a genetic defect in the renal vascular bed, a defect closely associated with the hypertensive trait. Atherosclerotic renal artery stenosis is a major, potentially remediable cause of chronic renal failure, especially in whites. Its prevalence in the atherosclerotic population is in the order of 15 percent. This figure has obvious bearing in terms of health cost. Early diagnosis and treatment by angioplasty or surgery can preclude development to end-stage renal disease and maintenance hemodialysis, as renal atrophy due to chronic ischemia resulting from renal artery stenosis can be halted or partially reversed by revascularization before extensive fibrosis sets in. Finally, renal vascular lesions are commonly observed in the course of various nephropathies, even in the absence of hypertension. The relationship between fibrogenesis and these vascular lesions, which develop along with interstitial fibrosis and entail an unfavorable prognosis in various glomerulopathies, remains to be elucidated. This is especially the case for focal-segmental glomerulosclerosis, membranous glomerulopathy and IgA glomerulonephritis. The pathophysiology of renal fibrosis induced by ischemia is centered on increased generation of angiotensin II that is fibrogenic owing to interaction with endothelin 1, PDGF-BB and TGF-beta. These notions open perspectives toward pharmacologic means to retard or even prevent the development of such various ischemic conditions to end-stage renal failure. PMID- 10371765 TI - [Genetics of kidney diseases and renal fibrosis]. AB - Renal fibrosis is found in genetic kidney diseases. In some of them, fibrosis may be due to mechanisms not dependent on the gene defect. In other cases, fibrosis depends more specifically on the gene defect: e.g. in juvenile nephronophthisis where interstitial fibrosis predominates, or in Alport's syndrome where glomerulosclerosis is triggered probably by unbalanced distribution of alpha chains of type IV collagen in glomerular basement membrane. Study of genes predisposing to renal fibrosis and progression is more complex. Genes regulating the renin-angiotensin system have been considered. The DD polymorphism of the converting enzyme gene may be associated with a more rapid progression rate of renal failure in glomerular and non-glomerular diseases. This remains, however, to be confirmed in more appropriate studies. PMID- 10371764 TI - [Metabolic mechanisms of renal fibrosis: diabetes]. AB - Diabetic nephropathy, a cause for renal fibrosis, results from glomerular haemodynamic abnormalities: intra-glomerular hypertension is provoked by pre glomerular vasodilatation (a consequence of hyperglycaemia) facing to constitutive, post-glomerular, vascular resistances. Pre-glomerular vasodilatation is due to abnormal glucose metabolism during hyperglycaemia. Studies on pathophysiology, genetics, and treatment of diabetic nephropathy are oriented by these latter two components of risk for renal fibrosis. PMID- 10371767 TI - [A priority mission: prevention, early detection, identification of risk factors of progression of chronic renal insufficiency]. AB - The prevalence of chronic renal failure is continuously increasing. The reasons are the lengthening life expectancy of the general population, the increasing age of the patient population benefitting from renal replacement therapy and the growing number of vascular and diabetic nephropathies leading to end-stage renal disease. The financial burden of treating renal failure is considerable, estimated at nearly 2 percent of the national health budget. A priority for French departments of nephrology is to initiate and participate in epidemiological studies on renal failure in order to eradicate the present pitfalls: lack of exhaustiveness and non-validation of information. The need is to create networks for patient care and management, grouping specialists such as cardiologists, diabetologists, rheumatologists and gerontologists, who face the problems stemming from renal failure, for early identification of problems, in order to apply appropriate measures to slow disease progression and to initiate renal management at the optimal time. Active participation in research is indispensable, not only in multicentric studies but also in "nephrosurveillance" and for development of guidelines for good clinical practice and good prescription practice. Many tools are available for undertaking varied experimental research. This research can lead to identification of the cellular and molecular mechanisms underlying nephron destruction and to development of effective strategies to slow or halt the progression of lesions. Identification of genetic factors that favor the appearance or progression of renal failure is in its early stages and will continue to develop. All these efforts require communication and teaching. A last point: the need to revamp hospital organization to better respond to the needs of patients requiring only a few hours in specialized units. PMID- 10371766 TI - [Factors of progression and chronic renal insufficiency and their prevention]. AB - End-stage renal failure is a major cause of health-related expenditures in the Western countries. It can be prevented in many instances by therapeutic intervention first settled in animal models. Murine experimental models of nephron loss have led to the identification of several factors involved in renal disease progression. They include increased glomerular capillary hydrostatic pressure, glomerular hypertrophy, high blood glucose, high-level proteinuria, hypoxia, and genetic factors. The role of glomerular capillary hypertension and hypertrophy cannot, however, be established with certainty in man. Clinical studies have shown that male gender, increased systemic blood pressure, severe urinary albumin loss, type of initial nephropathy, diabetes, and genetic factors, all accelerate progression of renal disease to end-stage renal failure. It is now feasible to halt the progression of renal insufficiency. Clinical trials have demonstrated the beneficial effects of angiotensin converting enzyme inhibitors, particularly in those patients with high-level proteinuria, and of blood glucose control in diabetic patients. The place that restricted protein diet and lipid lowering intervention should take, remains debated. Further clinical trials aimed at testing the efficacy of new drugs including endothelin receptor antagonists are warranted in those patients with vascular or tubulo-interstitial renal diseases. PMID- 10371768 TI - [The missions of a Nephrology Department in the year 2000]. AB - The treatment of acute renal failure: 1) Evaluate new methods of dialysis that should be less aggressive than hemodialysis, such as hemodiafiltration, in order to decrease the hemodynamic risks of an elder population. 2) Improve teaching: education of students and of physicians at the University must be more effective to allow a better diagnosis of acute renal failure and therefore its prevention, and a rapid histological examination of renal tissue that constitutes the basis of etiopathogenic treatment of glomerular and/or vascular nephropathies. The treatment of chronic renal failure: 1) Improve and develop the present therapy: automated peritoneal dialysis at home should be developed. 2) The evaluation of new techniques of hemodialysis in the patient can only be undertaken at the University Hospitals. New experimental procedures can endow the artificial kidney with most of the metabolic functions of normal kidney. This method has not been tested yet in men. 3) The xenografts are very interesting approaches because of the lack of organs, however there are major obstacles including insufficient knowledge of the mechanisms of hyperacute graft rejection, of the possibility of transmission of anthropozoonoses from animals to humans, and of gene mutation of virus that usually are not pathogenic in man; finally studies on the adaptation of the xenograft, which possess different biological parameters, to the human milieu, should be considered. 4) Cell therapy or gene therapy are currently under investigation. The major difficulties are to assess that the vector used to transfer the gene is absolutely harmless to man and to restrict the gene delivery to the cells of interest. The molecular genetics determinants should be essential to prevent kidney diseases because they would help in defining the individual susceptibilities to develop nephropathies. PMID- 10371769 TI - [Role of endovascular treatments in the management of arteriopathies of the aged subject]. AB - Endovascular procedures have deeply modified the treatment of patients with peripheral arterial diseases. However risks of transluminal angioplasty are increased with patient age. In patients with acute ischemia by distal embolism or in situ thrombosis percutaneous thromboaspiration is an alternative to the Fogarty catheter. In patients with chronic critical ischemia, revascularization is necessary. Risks of surgical treatment are rather in favour of endovascular procedures. In patients suffering of intermittent claudication, results of a series of 38 patients confirm the efficacy of angioplasty. Thus endovascular procedures can be proposed in alternative to medical treatment to patients over 70 years of age. PMID- 10371770 TI - [Vaginal hysterectomy for non-prolapse uterus]. AB - Vaginal hysterectomy in the treatment of non prolapsed uterus without malignant disease is a well codified technique. Per operative complications, post operative morbidity are less frequent and less serious than by abdominal surgery and convalescent time is less extended. Financial cost is lower compared with laparoscopic hysterectomy. The successful result of the vaginal way is determined by anatomic factors: volume and mobility of the uterus, accessibility of the vagina, but above all by the training of the surgeons. A rate of seventy per cent of hysterectomies performed by the vaginal way in the case of this indication is a realistic aim in a hospital center. PMID- 10371771 TI - [Fronto-temporal degenerative dementia. A modern neuropathologic approach]. AB - The classification of degenerative dementias with fronto-temporal atrophy has been debated since the description of Pick's disease. The study of a clinico pathological series of 10 cases using immunohistochemistry lead to the following conclusions: reserving the name of Pick's disease to those cases with argyrophilic inclusions, the most recognisable and characteristic marker at neuropathological examination, allows an easy and reliable diagnosis; keeping on with the splitting of these disorders into various clinico-pathologic entities seems today more useful than grouping them into a single syndrome until new data, based for example on genetic analysis, show that different phenotypes correspond to the same disease. PMID- 10371772 TI - [Epidemiology of Alzheimer's disease: the Paquid experience]. AB - The Paquid research program was specifically designed to study the epidemiology of Alzheimer's disease (AD). Paquid is a prospective study of a representative random sample of 4,134 elderly people living in Gironde and Dordogne, two administrative areas around Bordeaux in South-Western France. This program allowed to obtain prevalence, incidence, survival of AD and to analyze risk factors, consequences and premonitory signs of this disease. PMID- 10371773 TI - [Early nuclear modifications related to cellular activation: from histology to molecular cytopathology (1967-1997)]. AB - The nuclear chromatin structure and function are altered as soon as the first steps of cellular activation induced by membrane stimulation. Different studies carried on lymphatic tissues as well as isolated cell suspensions (lympho monocytes) demonstrate that an early and transitory chromatin dispersion, visualised by nuclear refringency (1967), is related to the genome derepression allowing DNA transcription mechanism. This is linked to the expression of c-fos proto-oncogene and that of specific proteins synthesis. This occurs before c myc expression and could play a role in the regulation of surface antigens expression. Independent from the induction of cell proliferation, but related to cell differentiation, the early and transitory chromatin dispersion as well as the influence of c-fos related to proteic system activator (AP1) is discussed in referring to recent studies (1997), as well as the regulation factors of the chromatin filament structure. PMID- 10371774 TI - [At the crossroads between B lymphoproliferative syndromes to autoimmune diseases are CD5-positive B-lymphocytes: a new hypothesis]. AB - Cells of most of the chronic lymphoid leukemias of the B cell lineage express the CD5 molecule, and this CD5 + B cell subset may be expanded in nonorgan-specific autoimmune diseases. Since autoimmune features are common in lymphoproliferative disorders and the latter be a complication in autoimmune settings, CD5 + B cell may stand at the crossroads of these two conditions. Our hypothesis is, therefore, that there are differences in CD5 + B cells arising during ontogeny ("classical") and those whereby CD5 is induced by various stimuli ("induced"). There is also evidence to support a role for CD5 on B cells, sustaining the proliferation of activated B cells, whereas resting B cells undergo apoptosis. PMID- 10371775 TI - [Evaluation of the results of organ transplantation in France: is there a center effect?]. AB - The evaluation of the results of organ allografts for each type of organ and for each center is one of the prioritary missions of the Etablissement francais des Greffes (EfG). The objectives, the methodology and the results of this evaluation have been defined and discussed with all the organ transplantation teams, after a preliminary work of the Conseil Medical et Scientifique of the Etablissement. This paper describes the evaluation experiment conducted by the EfG between 1995 and 1998. The main objectives of this first phase of the evaluation project are the identification of centers with outside of the norm results and the study of the relationship between the number of transplants performed by each center and the quality of their results. The chosen quality indicators are the excess in the patients mortality rate, computed one year after the first transplantation, for vital organs, and in the one year kidney graft loss rate for kidney transplantation. The excess of mortality is defined as the difference between observed and expected mortality rates. The expected mortality rate is estimated, for each program, by a statistical model based on a set of patient specific risk factors. All the vital organ transplantation teams who have performed more than 10 transplants between 1991 and 1995, and all the kidney transplantation teams who have performed more than 15 transplants between 1991 and 1996 have been included in the study. The main results of this evaluation experiment are the following ones: even if the intercenter variability of the results was statistically significant, it remained of low magnitude, particularly for heart, liver and kidney transplantation. None of the evaluated centers presented results outside of the norm. The results were positively correlated to the number of transplants for liver and lung transplantation. This was not the case for heart and kidney transplantation; this relationship is difficult to analyze for heart lung transplantation, due to the small number of centers included. Thanks to the data base constituted since 1959 by all the organ transplantation teams, and in spite of the partially retrospective nature of this study, which explains its limits, this evaluation experiment, opens a perspective of extension to other domains of public health. In the future, however, this kind of evaluation should be prospective; a project aimed to developing the evaluation of the results of organ transplant actions in real time was defined and is currently on-going. PMID- 10371776 TI - [Apoptosis and cancer]. AB - Apoptosis or programmed cell death is a form of regulated cell death that represents an important biological principle in tissue development and homeostasis. Its regulation appears to be perturbed in several major diseases, including cancer. The control of cell death events is extremely complex at the genetic and biochemical levels. It has been suggested that genes or factors that suppress apoptosis are essential components of carcinogenesis. The complete mechanisms implicated in apoptosis remains an issue for a future investigation. Details of how the death pathway itself is initiated, will provide a firm basis for the development of agents capable of eliciting the process in chemoresistant tumours. PMID- 10371777 TI - [Genotype, serotype and sensitivity to fluconazole of Candida albicans strains isolated from HIV-positive patients]. AB - Genotype, serotype and susceptibility in vitro to fluconazole of 104 C. albicans strains isolated from HIV+ patients were studied. The possible correlations between genotype analysed by multilocus enzyme electrophoresis (MLEE) and phenotype of medical relevance (serotype and susceptibility to fluconazole) of Candida albicans isolated from these patients treated with fluconazole were evaluated by factorial correspondence analysis. No correlation was observed between genotype and in vitro or clinical response to fluconazole. In counterpart, serotype B C. albicans was associated with some multilocus genotypic patterns. PMID- 10371778 TI - [Neuronal apoptosis in human prion diseases]. AB - Neuronal loss is a salient feature of prion diseases; however, its causes and mechanisms are unclear. The possibility that it could occur through an apoptotic process has been postulated and this is consistent with the lack of inflammation in prion disorders as supported by experimental studies. In order to test this hypothesis in humans, we examined samples of frontal and temporal cerebral cortex, striatum, thalamus and cerebellum from 26 patients who died from prion diseases. They included 16 cases of Creutzfeldt-Jakob disease (5 sporadic cases, 5 familial, 3 iatrogenic, and 3 cases with the new variant), and 10 cases of fatal familial insomnia including 8 homozygotes methionine/methionone at codon 129 of the prion protein gene and 2 heterozygotes. These were compared with age and sex matched controls. Using in situ end labelling, we identified apoptotic neurons in all the cases of Creutzfeldt-Jakob disease. A single labelled neuron was found in the eldest control. Apoptotic neurons were mostly found in damaged regions and their presence and abundance seemed to correlate closely with neuronal loss. This supports the view that apoptosis of neurons is a feature of prion diseases and may contribute to the neuronal loss which is one of the main characteristics of these conditions. Neuronal apoptosis also correlated well with microglial activation as demonstrated by the expression of major histocompatibility complex class II antigens and axonal damage as identified by beta-amyloid protein precursor immunostaining. In contrast, we found no obvious relationship between the topography and severity of neuronal apoptosis and the type, topography and abundance of prion protein deposits as demonstrated by immunohistochemistry. PMID- 10371779 TI - [Exterior and interior air pollution. Its impact on health]. PMID- 10371780 TI - [Relationships between air conditioning, airborne microorganisms and health]. AB - Concurrently with the increase of air-conditioning, potentially severe or frequent new diseases have emerged, giving rise to social and economical consequences. The first part of this work is a state of the art review of the relationships between air-conditioning, airborne microorganisms and health, through a technical, metrological and medical approach. The second part presents four studies performed in this field. Two of them deal with the relationship between airborne microorganisms and technical features of air-conditioning. Measurements performed on actual sites demonstrated the benefit of using high efficiency filters and low risk components in air-conditioning systems. The third study was aimed to look for a relationship between airborne microorganisms and sick building syndrome symptoms. Statistical analyses of individual data revealed significant associations between airborne bacteria or fungi and symptoms. These results may be the first step in determining a dose-response relationship, in order to define threshold limit values in this field. In the fourth study, the contribution of particle counting in assessing exposure to airborne microorganisms was explored by monitoring simultaneous variations of microbial and particle concentrations. The results showed that associating particle counting may allow to detect microbial variations instantaneously, and therefore improve the assessment of exposure to airborne microorganisms. PMID- 10371781 TI - [Cellular pathophysiologic effects of various air pollutants on the airways]. AB - Both epidemiological and experimental data indicate that major health consequences of urban air pollution i.e., respiratory symptoms are related, at least in part, to an alteration in the cellular processes implicated in bronchial responsiveness. In the Laboratory, we have set up techniques which enable to pre expose in vitro human isolated bronchi to some air pollutants. Using these techniques, we have examined the dosimetric relationships and the cellular mechanisms of bronchial responsiveness altered by nitrogen dioxide (NO2), ozone (O3) or aldehydes such as acrolein. The combined effects of gas pollutants and passive immunological sensitization on bronchial hyperresponsiveness have also been studied. As a general rule, experimental protocols consisted in a comparison of the responsiveness of human bronchial tissues (obtained at thoracotomy from patients undergoing resection for pulmonary carcinoma) following ex vivo exposure to air pollutants with that of paired tissues unexposed to pollutants which acted as temporal controls. We have observed that the responsiveness of human isolated bronchi is increased following pre-exposure to NO2 or O3 or to acrolein for 15 to 30 min, at concentrations in the range of ppm and microM, respectively. A common target for the action of these pollutants on airway smooth muscle reactivity has been identified i.e., the release of intracellular stored calcium ions. Direct measurements of cytosolic calcium concentration in isolated airway smooth muscle cells exposed to pollutants have confirmed this hypothesis. Finally, we have obtained results indicating that passive sensitization and exposure to pollutants act in a additional manner on human bronchial smooth muscle reactivity in response to both specific antigen and non specific agonists. Collectively, these experimental in vitro results enable (i) to establish dosimetric relationships, (ii) to examine the cellular mechanisms and (iii) to identify populations at risk for various gas pollutants. PMID- 10371782 TI - [Teledermatology--importance of telemedicine and dermatology]. AB - Due to a constantly improving technology the transfer of clinical images associated with histological images represents the actual answer to the expectation of Dermatologists. The authors give their personal experience regarding the use of telemedicine in Dermatology: discussion of clinical cases, histopathological presentation, diagnostic assistance, follow-up of patients, teaching, and research. Perspectives and development offered by telemedicine as well as the advantages of patient care, and of presenting research topics are mentioned. Also aspects which have to be clarified before telemedicine takes up are presented: as far as that goes for practicing Medicine and Dermatology, technical applicability, credibility, acceptance, confidence and responsibility. PMID- 10371783 TI - [Prevention of aspergillosis with itraconazole in neutropenic patients: importance of drug monitoring]. AB - Itraconazole is being used increasingly as prophylaxis of systemic aspergillosis in patients with immunodepression. Therapeutic itraconazole monitoring by plasma concentrations measurement is justified by its dose-dependent pharmacokinetics, drug interactions, and frequent bioavailability modifications observed in immunocompromised patients. A first study was carried out in 16 patients with haematological malignancies, given chemotherapy plus itraconazole 400-800 mg/day in a single dose as prophylactic treatment. Therapeutic through drug plasma concentration (Cmin > or = 250 ng/ml of itraconazole, or > or = 750 ng/ml of itraconazole plus hydroxyitraconazole) was not reached in 5/16 patients. Moreover results emphasised the wide inter and intra-individual variability of the steady state plasma concentrations. In another study we used a multiple dose regimen (100-200 mg x 3/day), instead of a single dose regimen. An adequate Cmin value was found in 34/36 patients. In conclusion, the wide inter and intra-individual variability of itraconazole pharmacokinetics necessitate regularly to measure the drug plasma concentrations in patients with life-threatening fungal infections. PMID- 10371784 TI - Directional coronary atherectomy: from therapeutic device to research tool in coronary artery disease. AB - Directional coronary atherectomy (DCA) was introduced as a new percutaneous revascularization modality in 1990, and was initially applied to large vessels without tortuosity or calcification, with overall results including a 95% procedural success, 94% clinical success and 4.6% major complications (urgent bypass surgery in 3.8%, Q wave myocardial infarction in 1.7%, and hospital mortality in 0.3% of patients). In addition to its established efficacy for eccentric lesions, newer applications emerged such as treatment of saphenous vein grafts, thrombus-associated lesions, aorto-ostial lesions, failed or suboptional coronary angioplasty results, bifurcation lesions and use as a part of multi vessel intervention. Comparative studies with coronary angioplasty such as CAVEAT I and II and CCAT showed better success rates with DCA vs coronary angioplasty, but failed to demonstrate benefit in restenosis rates. OARS and BOAT studies helped define optimal atherectomy techniques, which led to better acute angiographic results and to the "debulking plus stenting" concept. A spin-off of those clinical applications has been the opportunity to study the histology of tissue excised by DCA in vivo in different clinical settings. Such studies, investigating plaque ulceration, thrombosis and inflammation are reviewed, with special emphasis on new insights into unstable angina; the future of atherectomy research is also outlined with a categorization of various possible protocols to be applied utilizing coronary atherectomy specimens from live patients. PMID- 10371785 TI - Homocysteine and arterial occlusive disease: a concise review. AB - Many cross-sectional and prospective studies have shown that raised serum/plasma levels of total homocysteine increase the risk of coronary, cerebral, and peripheral artery disease. The risk associated with hyperhomocysteinemia appears to be concentration-dependent and not attributable to traditional risk factors. The odds ratio for ischemic heart disease has been estimated to be 1.4 for every 5 mumol/l increase of total plasma homocysteine. Median fasting total plasma homocysteine in adult males is approximately 10 mumol/l. Mild hyperhomocysteinemia is estimated to occur in 5-10% of the general population. Plasma concentrations are increased as a result of age, male gender, impaired renal function, low vitamin B intake, and genetically-determined defects of the enzymes involved in homocysteine metabolism. Folate supplements can reduce total homocysteine levels by approximately 25%. Studies in vitro and in vivo indicate that homocysteine can impair endothelial function. Despite increasing recognition of hyperhomocysteinemia as a risk factor for arterial occlusive disease, irrefutable proof that mild hyperhomocysteinemia contributes directly to the pathogenesis of atherothrombosis will come if interventions to lower total homocysteine reduce cardiovascular events. Family studies may also provide evidence of causality if genetic causes of hyperhomocysteinemia are found to segregate with disease. PMID- 10371786 TI - Genetics and cardiovascular risk: a role for apolipoprotein(a) polymorphism. AB - Apolipoprotein(a) [apo(a)] is the specific apolipoprotein of lipoprotein(a) [Lp(a)], a recognized cardiovascular risk factor. Apo(a) is characterized by a high genetic polymorphism with at least 34 isoforms in plasma. Recent studies have shown that in atherothrombosis apo(a) polymorphism could play a role independent of Lp(a) levels. In particular, apo(a) phenotypes seem to have their highest predictive value for coronary heart disease, when apo(a) isoforms are detected by high resolution phenotyping methods and when an adequate operative cut-off of apo(a) polymorphism is used. A strong association between apo(a) phenotypes and coronary heart disease has been also found in hypertensive, diabetic, and uremic patients. Moreover, apo(a) phenotypes seem to correlate well with the severity of coronary atherosclerosis and the age of clinical onset of coronary heart disease. These studies suggest that apo(a) polymorphism may have a great clinical usefulness in a primary prevention setting, since apo(a) phenotypes could be used together with Lp(a) levels as strong genetic predictors of atherothrombosis. The analysis of apo(a) polymorphism appears to be particularly useful in healthy subjects with a family history of atherothrombotic diseases, in patients with diseases at high cardiovascular risk (diabetes, hypertension, hypercholesterolemia) and in subjects with conditions modifying Lp(a) levels. PMID- 10371787 TI - [Heterogeneities of ventricular repolarization and vulnerability to arrhythmia. How to detect them with noninvasive methods?]. AB - Vulnerability to arrhythmias can be influenced by two conditions: a dynamic (beat to-beat) variation of repolarization sequence, and a state of heterogeneity of repolarization, i.e. a greater than normal dispersion of recovery time. The first condition is well reflected by T-wave alternans, a phenomenon characterized by alternation on every other beat basis of amplitude and morphology of T waves. Experimental studies provided evidences of close temporal correlations between ischemia-induced alternans, dispersion of repolarization and susceptibility to ventricular fibrillation. Gross T-wave alternans can be occasionally observed in patients with long QT syndrome or during acute ischemia before the onset of arrhythmias. Recent studies have demonstrated that measurement of microvolt level T-wave alternans at rest and during exercise is a promising technique for the identification of patients at risk of ventricular arrhythmias and sudden death. A state of repolarization inhomogeneity can be revealed by methods which analyze a single cardiac beat. The QT dispersion, defined as the difference between maximum and minimum QT interval measured at 12 lead ECG, is the most simple and widely used index of repolarization inhomogeneity. The major limitation is that this measure cannot be related to the actual spatial heterogeneity of repolarization, since each surface lead reflects, in different degree, the electrical activity of the whole heart. The majority of studies reported that, in various pathological conditions, the QT dispersion is higher in patients with than without ventricular arrhythmias. On the other hand, a recent large prospective study in post myocardial infarction patients failed to demonstrate the predictive value of QT dispersion, even when measured with the best available methodology. Body surface potential mapping has proven to be a useful method for detecting repolarization inhomogeneities not revealed by the analysis of conventional ECG leads. Different methods of analysis of the potential maps have been used. By applying principal component analysis of the ST-T waves, we computed the similarity index, defined as the ratio of the first principal component to the sum of all remaining components. A low value of similarity index suggests a high degree of repolarization inhomogeneity. The similarity index was found significantly lower in patients with idiopathic long QT syndrome and in patients with arrhythmogenic right ventricular dysplasia with episodes of ventricular tachycardia than in normal subjects. Future researches should aim at identifying novel reliable indices of repolarization inhomogeneity, first deduced from extensive body surface mapping, then possibly computed from digital recording of the 12 conventional leads. PMID- 10371788 TI - Sick sinus syndrome with and without atrial fibrillation: atrial refractoriness and conduction characteristics. AB - BACKGROUND: Clinical electrophysiology has focused the attention on the electrophysiological properties of the atrial muscle in patients with atrial fibrillation: shortened and inhomogeneous refractoriness and local and regional conduction slowing, as well as prolonged intra- and interatrial conduction disturbances, are well described as electrophysiological parameters associated with the genesis of atrial fibrillation. Patients with sick sinus syndrome are variously included in these studies, but electrophysiological characteristics of patients with sick sinus syndrome alone appear less investigated, even if atrial fibrillation is part of its natural history. The aim of the present study was to define the electrophysiological characteristics of sick sinus syndrome patients with or without paroxysmal atrial fibrillation, compared to subjects without atrial fibrillation and sick sinus syndrome. METHODS: We reviewed the electrophysiological data of 39 patients with sick sinus syndrome (mean age 70 +/ 8 years), who underwent an electrophysiological study in sinus rhythm for the evaluation of the atrial substrate. In 12 patients an associated history of paroxysmal atrial fibrillation was documented. Twenty-seven patients were included in the study with a diagnosis of sinus node dysfunction alone. We also considered as control group 25 subjects (mean age 63 +/- 14 years), referred to our electrophysiological laboratory for unexplained syncope or atrioventricular disturbances. Following pharmacological wash-out and at a drive cycle of 600 ms, effective and functional refractory periods, S1-A1 and S2-A2 latency, A1 and A2 width, and the latent vulnerability index (effective refractory period/A2), were measured. In addition, the P-wave duration during spontaneous sinus rhythm on the surface ECG in D II/V1 leads was measured. RESULTS: Between sick sinus syndrome patients with or without atrial fibrillation, no significant statistical differences in electrophysiological parameters were found. When compared to the control group, sick sinus syndrome patients did not show any differences in effective refractory period (239 +/- 34 vs 250 +/- 29 ms), functional refractory period (276 +/- 28 vs 280 +/- 32 ms), S1-A1 (38 +/- 16 vs 33 +/- 11 ms), and S2 A2 latency (68 +/- 25 vs 63 +/- 25 ms). In contrast, we observed remarkable differences in terms of atriogram duration A1 (60 +/- 20 vs 39 +/- 13 ms, p < 0.001), A2 (95 +/- 34 vs 57 +/- 18 ms, p < 0.001), and effective refractory period/A2 (2.8 +/- 1.2 vs 4.8 +/- 1.7 cm, p < 0.001). Also the duration of the P wave was longer (103 +/- 17 vs 94 +/- 45 ms, p < 0.05). CONCLUSIONS: In sick sinus syndrome patients with or without atrial fibrillation, electrophysiological characteristics appear homogeneous. When compared to the control group, refractoriness was quite similar. In contrast, the most important abnormalities appear based on conduction slowing disturbances, responsible for a low latent vulnerability index. This could explain, at least in part, the tendency of sick sinus syndrome to develop atrial fibrillation as a part of its natural history. At present, the influence of an altered electrophysiological substrate on pharmacological or pacing therapy in patients with sick sinus syndrome is not yet known. PMID- 10371789 TI - [Restoring the mechanical atrial function after cardioversion of atrial fibrillation: clinical and echocardiographic predictive factors]. AB - BACKGROUND: A delay in the recovery of effective mechanical atrial function after cardioversion for atrial fibrillation can predispose to thromboembolism. The aim of the present study was to assess the influence of clinical and echocardiographic parameters on the recovery of left atrial contraction after cardioversion of atrial fibrillation. METHODS: One hundred and 36 consecutive patients were evaluated and 80 were randomly cardioverted using either DC shock or i.v. procainamide. Patients who recovered sinus rhythm (26 patients treated with procainamide and 39 patients cardioverted with DC shock) underwent a complete Doppler echocardiographic examination 1 hour after the restoration of sinus rhythm and after 1 and 7 days and 1, 3, and 6 months. The following parameters were evaluated: age, underlying cardiac disease, duration and etiology of atrial fibrillation, mode of cardioversion, left ventricular diameters and function, left atrial diameters and function, assessed as atrial ejection force. The relation between these variables and atrial ejection force was tested. RESULTS: Atrial ejection force was greater immediately and at 24 hours after cardioversion in patients who underwent pharmacological therapy compared to patients treated with DC shock. In all groups atrial ejection force increased over time. The mode of cardioversion was significantly associated with the recovery of left atrial mechanical function by day 1 in univariate and multivariate analyses (odds ratio 0.14, 95% confidence interval 0.03-1.6). The other variable associated with atrial ejection force was left atrial size (odds ratio 0.15, 95% confidence interval 0.17-1.9). CONCLUSIONS: Atrial ejection force can be easily measured after cardioversion to obtain accurate information about the recovery of left atrial mechanical function. In the present study the recovery of left atrial function was influenced by the mode of cardioversion and left atrial size. PMID- 10371791 TI - [Remodelling of the radial artery graft 5 years after aortocoronary bypass intervention]. AB - BACKGROUND: The radial artery (RA) is being employed as coronary artery bypass graft with good results, but when it is proximally anastomosed to the ascending aorta, undergoes substantial hemodynamic changes which could lead to significant graft intimal hyperplasia. The aim of this study was to investigate the evolution of RA graft morphology over time. METHODS: We studied 20 patients with a perfectly patent RA graft at both 1 and 5 year angiography after coronary artery bypass graft. RESULTS: Both RA graft and grafted coronary artery diameters, assessed by quantitative coronary angiography, significantly increased at 5 years, in comparison to 1 year angiography (2.08 +/- 0.45 vs 2.54 +/- 0.53 mm, +22%, p < 0.001 and 1.92 +/- 0.47 vs 2.18 +/- 0.41 mm, +13.3%, p < 0.001, respectively). CONCLUSIONS: Hemodynamic changes following RA employment for coronary artery bypass graft stimulate a remodeling of RA graft itself and of the grafted coronary arteries. The progressive increase of diameters observed in RA grafts strongly argues against the development of flow-limiting graft intimal hyperplasia when RA is proximally anastomosed to the ascending aorta. Moreover, grafted coronary artery dilation suggests that hemorrheologic changes following coronary artery bypass graft could play a major role in the development of RA remodeling over time. PMID- 10371790 TI - In vitro hyperreactivity to lipopolysaccharide in patients with history of unstable angina is not associated with seropositivity for Cytomegalovirus, Helicobacter pylori and Chlamydia pneumoniae. AB - BACKGROUND: Inflammation and possibly chronic infections are associated with acute coronary syndromes; however, the mechanisms responsible for this association are not yet fully elucidated. The aim of this study was to assess whether the hyperreactivity of the inflammatory system, that we have shown in unstable patients with persistently elevated C-reactive protein and with recurrence of symptoms, was associated with chronic infection. METHODS: In 20 unstable angina patients seropositivity and antibody levels vs Cytomegalovirus, Helicobacter pylori and Chlamydia pneumoniae were measured and correlated with the interleukin-6 production in vivo in 1 ml of whole blood stimulated with 0.1 microgram lipopolysaccharide for 4 hours. RESULTS: No positive correlation was found between antibody titer and interleukin-6 levels. No correlation was also found between seropositivity to Cytomegalovirus, Helicobacter pylori or Chlamydia pneumoniae and interleukin-6 levels. CONCLUSIONS: Our study suggests that seropositivity for infective agents, including Chlamydia pneumoniae, does not affect the monocyte response to lipopolysaccharide and thus cannot account for the enhanced interleukin-6 production observed in unstable angina patients with raised levels of C-reactive protein and worse prognosis, and suggests the predominant role of the individual response to different stimuli. PMID- 10371792 TI - [Single coronary artery with a right origin: angiography identification of the R II subgroup]. AB - This case report illustrates the clinical and angiographic findings of 2 patients undergoing coronary angiography for ischemic heart disease and with the unexpected presence of anomalous origin of the left coronary artery from the right aortic sinus. The angiographic classification of the different subgroups of single coronary artery is reviewed and the 2 cases are identified as type R-II with septal and anterior course of the left main stem. PMID- 10371793 TI - Pneumothorax and conventional ventilation in the neonatal period. AB - BACKGROUND: Pneumothorax (pnth) as a form of the air-leak syndrome cannot be fully prevented during conventional and nonconventional ventilation. Therefore, all efforts targeted at elucidation, prognosis and prevention or decrease of the air-leak syndrome are important and essential. The objectives of the present study were to study the dynamics of the panel of indices based on the ventilator parameters and blood-gas analysis in newborns and premature neonates with pneumothorax treated with conventional ventilation who subsequently developed pneumothorax, to compare the results in survived with those of nonsurvived, to compare the computer models in the patients enrolled in the present study with the results presented in our previous investigations and to determine their clinical and prognostic significance for early detection of impending pneumothorax, the choice of optimal ventilatory technique, as well as their place with regard to the early and later evolution in the condition of the newborn. PATIENTS AND METHODS: A total of 22 newborns (10 survived and 12 nonsurvived, born in term and prematurely) who had been treated with conventional ventilation and subsequently developed pneumothorax were enrolled in the present study. The primary data (clinical and laboratory-blood-gas analysis, ventilatory parameters and their derivative indices--MAP, OI, VI, a/A, VEI, AaDO2), were calculated using the Neonatal Intensive Computer File. RESULTS AND DISCUSSION: We followed prospectively the dynamics of the panel of indices and compared the values and the major trends between the survived and the nonsurvived with pneumothorax as well as the computer models of the dynamics of the indices in the neonates with pneumothorax and the general pattern for the respective group as presented in our previous studies. Another important point in the comparative discussion is the significant difference in the values of R2. CONCLUSIONS: 1. The dynamics of the indices that we followed shows some characteristic findings and differences between the survived and nonsurvived newborns with pnth during the first three days following the initiation of conventional ventilation. 2. We found a significant difference in the values and dynamics of the indices between the newborns with pnth and those from the general groups. 3. Despite the fact that the predictive value of the panel of indices in newborns with pnth is lower as compared with the general groups, the interactions have important clinical implications for the management of artificial ventilation in high-risk newborns. PMID- 10371794 TI - Local effects on the blood gas indices in diaphyseal forearm fractures. AB - We studied the local changes of blood gas indices of arterialised capillary blood taken simultaneously from the fractured and the uninjured arm of 30 patients with diaphyseal fractures of the forearm. Blood gases were measured with a Stat Profile 5 analyser (Nova Biomedical). The results indicate a slight but statistically significant decrease of the acid-base indices without any changes in the oxygen indices. These slight changes do not affect considerably the diagnosis of the profile. PMID- 10371795 TI - Peptic esophageal stricture in children. AB - INTRODUCTION: Peptic esophageal stricture as a complication of gastroesophageal reflux disease (GERD) occurs in 5% of the affected children. MATERIAL AND METHODS: Case histories of 6 children treated successfully in the Department of Pediatrics and Clinic of Pediatric Surgery were studied. The diagnosis in each case was based on clinical symptoms (vomiting leading to hypothrophy, hematemesis, and anemia), and esophagoscopy, esophageal pH-metry (according to ESPGAN recommendations), and contrast X-ray examination. After evaluation medical treatment was applied in 3 and bougienage with a hard bougie in 6 patients. Because of failure of this treatment Nissen fundoplication and postoperative bougienage were performed in all patients. RESULTS: In all surgically treated patients complete recovery without postoperative complications was achieved. DISCUSSION: The authors give interpretation of the pathogenesis and outline the primary symptoms of the disease. Terms of performance and reliability of the instrumental methods of diagnosing are discussed. The experience in treatment of peptic esophageal stricture in children is presented. CONCLUSIONS: Medical treatment combined with bougienage yields poor results in peptic esophageal stricture and Nissen fundoplication appears to be the treatment of choice. PMID- 10371796 TI - Comparative drug monitoring of carbamazepin suspension (TroyaPharm) and Tegretol syrup (Ciba Geigy) in epileptic children. AB - INTRODUCTION: Administration of antiepileptic therapy requires use of proper drugs. The introduction and implementation of home carbamazepine preparations has substantial economic and social effects for Bulgaria. METHODS: Fifty nine epileptic children were enrolled in the trial. The serum concentrations of Carbamazepin suspension (TroyaPharm, Bulgaria) and Tegretol syrup (Ciba Geigy, Switzerland) were evaluated by gas-chromatographic analysis. RESULTS: Steady state serum antiepileptic drug concentrations (4-12 micrograms/ml) were maintained during the 3-month period of administration of both preparations. At the end of the first month of trial serum drug concentrations below the therapeutic range were observed in 15.2% of the patients on carbamazepin therapy and in 33.3% of the patients treated with Tegretol. At the end of the third month these indices changed to 21.2% and 13.3%, respectively. At the end of the first month serum level above the therapeutic range was found in 3.0% of the carbamazepin-treated patients and in none of those receiving Tegretol, while at the end of the third month in none of the blood samples did serum concentration of both drugs exceed the therapeutic range. Both preparations showed quick and substantial absorption in the first two hours after administration. It was determined by the indirect pharmacokinetic variable delta, representing the difference CSS2I- CssminI and expressed as a percentage of CssminI. This variable was 45.43% for Carbamazepin suspension and 41.00% for Tegretol syrup. CONCLUSIONS: No statistically significant differences in the examined pharmacokinetic parameters were found between both preparations. PMID- 10371797 TI - Clinical significance of serum HBeAg and HBV-DNA-specific values of virus replication in chronic hepatitis-B virus infection. AB - The prevalence of serum HBV-DNA and that of HBeAg was evaluated in 44 subjects (27 males and 17 females) aged between 3 and 59 years. They were divided in two groups: (A) chronic asymptomatic HBsAg carriers; and (B) chronic HBsAg carriers with a history of HBV infection. The patients had been chronic HBV carriers between 8 months and 15 years. All underwent clinical and biochemical evaluation. The serological markers of HBV infection were tested using ABBOTT assay kits. The serum HBV-DNA was quantified using a hemiluminiscence molecular hybridization assay (Digene-Murex). HBV-DNA+ were 13 patients (29.55 +/- 6.88%). The highest level of viral replication (up to 50%) was measured in the patients aged from 3 to 29 years while in the others a 3 to 4-fold decrease of the viral replication was detected. HBV-DNA+ were 8 (23.53 +/- 6.39%) of the chronic asymptomatic hepatitis B carriers and 5 (50.00 +/- 7.5%) of the chronic HBV carriers with former acute hepatitis B infection. Similar results were obtained for the other index of viral replication--HBeAg/anti-HBe. Eight (23.53 +/- 6.39%) patients from group I and 4 (40.00 +/- 7.38%) patients from group II were HBeAg+ while anti HBe+ were 26 (76.47 +/- 6.39%) and 6 (60.00 +/- 7.38%) patients from group I and II, respectively, i.e., about a quarter of the chronic asymptomatic HBsAg carriers and half of the chronic HBV carriers that had had an acute hepatitis B virus infection had HBV replication in their bodies. HBeAg+ patients had high levels of serum HBV-DNA (625.70-3328.00 pg/ml) which indicated extremely intensive viral replication. The presence of HBeAg and especially of HBV-DNA as markers of viral replication in chronic asymptomatic HBsAg carriers and chronic HBsAg carriers with a prior acute hepatitis B virus infection provide important information for the clinical decisions. PMID- 10371798 TI - Isolation and identification of Geotrichum candidum as an etiologic agent of geotrichosis in Bulgaria. AB - Geotrichosis affects mainly patients with systemic diseases like diabetes mellitus, leukoses, neoplasms etc. Clinically, it is similar to candidiasis and may occur as an oral, vaginal, skin, or systemic infection. Clinical specimens (98 sputa and 67 oral smears) were collected and studied using microscopic examination of Gram stained preparations and culture sampling between 1995 and 1997. Geotrichum candidum was isolated as a single pathogen in 8 sputum and 7 oral smear samples. Ten-day antifungal treatment with Nizoral was applied and resulted in relatively quick clinical improvement. The presented cases are the first cases of pulmonary and oral infections reported in our home practice in which Geotrichum candidum species was identified as a pathogen. The identification of Geotrichum candidum using combination of colonial and microscopic morphologic features increase the possibilities for diagnostic decision. PMID- 10371799 TI - A case of Chlamydia trachomatis infection in a renal allograft patient. AB - We describe a renal allograft patient with a Chlamydia trachomatis infection. A 43 year-old man was diagnosed with end-stage renal disease in 1985 which necessitated the transplantation of a cadaver kidney in 1986. The kidney was rejected two years later. A second transplantation was performed in 1991. At the beginning of 1998 symptoms and signs of chronic renal failure and dysuria set in. Routine microbiological studies were negative. Cell culture on McCoy cell line was positive for an active infection with C. trachomatis--initially 3+, then 2+, 1+ and negative following treatment. The patient was positive on the AMPLICOR CT/NG test (Roche Diagnostic Systems, Branchburg, USA) twice with OD values OVER- above 2 at 450 nm wavelength measured on an ELISA reader. The patient received treatment with azithromycin and doxycycline for 10 days following which the serum creatinine levels fell and the creatinine clearance values improved. Dynamic microbiological follow-up showed disappearance of C. trachomatis as evidenced by the negative PCR test. We conclude that the deterioration of renal function in our patient is complex but the infection with C. trachomatis is part of the complex of the underlying chronic renal failure and immunosuppressive treatment. PMID- 10371800 TI - Dynamic follow-up of the intact parathormone levels in hemodialysis patients treated with Rocaltrol and calcium. AB - The object of the present study was to follow prospectively the serum levels of intact parathormone (PTH) of hemodialysis patients and the subsequent changes following the oral administration of 1.25(OH)D3 and calcium. METHODS: We studied 30 chronic renal failure hemodialysis patients--16 men and 14 women, aged 20-70 years. Twenty-one of them were on hemodialysis with duration of up to 5 years (Group 1) and nine--up to 10 years (Group 2). All patients received oral supplementation therapy with 1.0 elemental calcium and Rocaltrol (Roche) 0.25 microgram/day. We measured the serum calcium, ionized calcium, serum phosphorus, alkaline phosphatase and the intact serum PTH levels in intervals of 12 months. RESULTS: Patients with duration of dialysis of up to 5 years had a significantly lower baseline PTH level of 392.5 +/- 94.7 pg/ml versus 896.4 +/- 160.7 pg/ml for those from the second group (P < 0.01). The intact PTH levels showed a tendency towards decrease--at the end of the study they were as follows: 372.02 +/- 76.9 for group 1 versus a significant increase for those from group 2--serum PTH levels of 1793.65 +/- 290.3 (P < 0.02). The differences in alkaline phosphatase and serum phosphorus levels at the end of the study period failed to reach statistical significance. Serum calcium levels were increased in both groups following the initiation of treatment but the difference was statistically significant only for group 2. A significant positive correlation was observed between the duration of hemodialysis treatment and the intact serum PTH levels. CONCLUSIONS: 1. Long-term low-dose conventional calcitriol therapy in combination with calcium supplementation could slow the progression of secondary hyperparathyroidism in some hemodialysis patients. 2. Low-dose therapy with active vitamin D-metabolites is effective only in hemodialysis patients with baseline serum PTH levels below 500 pg/ml and without pronounced hyperphosphatemia. PMID- 10371801 TI - A study on the prevalence of caries incipiens in 7-, 12- and 14-year-old children from Plovdiv. AB - INTRODUCTION: The decline in dental caries prevalence in many countries of the world in the last decades requires the adoption of new approaches towards determination of prevention strategies. The individuals and groups at high risk of developing caries should be differentiated from the rest of the population with the aim of carrying out selective prevention. Of particular interest is the determination of the risk of developing caries before it is clinically manifested as this would allow execution of a timely and adequate prevention. The aim of the present study was to determine the prevalence of caries incipiens on the teeth and on the dental surfaces in 7-, 12- and 14-year-old children and to compare it with the prevalence of clinical caries in the same population groups. METHODS: The study is representative by design and compares 600 children aged 7, 12 and 14 years from Plovdiv. The study is designed and carried out in compliance with the guidelines of WHO for conducting clinico-statistical study of dental caries. The diagnostic procedure of caries incipiens is visual-tactile. The test of vital staining with 2% water solution of Methylene Blau was also used. RESULTS: Caries incipiens prevalence was found to be higher than that of clinical caries in 7 year-old children (P < 0.001). In 12- and 14-year-old children the ratio of caries incipiens to clinical caries shifts in favour of clinical caries (P < 0.001). CONCLUSIONS: The higher prevalence of caries incipiens compared to clinical caries in 7-year-old children (P < 0.001) justifies its more extensive investigation as a predictive factor for developing caries in the 7-12 years age range groups. PMID- 10371802 TI - Prevention of dental caries on the first permanent molars with fluoride gel in the first year after eruption. AB - INTRODUCTION: The first permanent molars (M1) are especially sensitive to caries attack immediately after eruption. The questions concerning the effect of preventive methods during this period in groups at low risk of caries are insufficiently studied. The aim of the study was to examine the effect of caries prevention of M1 in the first year after eruption by means of fluoride gel in groups with low risk of caries. METHODS: A clinical study including 213 seven year-old children from Plovdiv was conducted in the course of one year. The children were assigned to two groups--experimental and control. In both groups children without clinical caries on M1 were included. The sampling procedure excluded subjects with multiple and diffuse obturations of the deciduous teeth, inadequate oral hygiene or undergoing orthodontics treatment. In the experimental groups four applications with 0.42% fluoride gel were performed at three months intervals. RESULTS: At the end of the study period the prevalence rate of caries on teeth and on dental surfaces was higher in the control group (P < 0.001). Caries reduction in the experimental group was 73.81% on the teeth and 73.12% on the dental surfaces. CONCLUSIONS: The results of our study show that immediately after eruption the first permanent molars are susceptible to caries in individuals at low risk of caries. This necessitates prevention activity during this period irrespective of the presumed risk. The fluoride gel applied by the author shows high anticaries activity on M1 in the first year after the eruption. PMID- 10371803 TI - Erythrocyte ferritin levels in chronic renal failure patients. AB - The authors studied the level of erythrocyte ferritin and its importance as a marker of iron metabolism in renal anemia patients. METHODS: We studied 33 chronic renal failure patients--12 men and 21 women, aged 33-72 years. Hemoglobin levels, erythrocyte counts, hematocrit, mean corpuscular hemoglobin, serum creatinine, iron and ferritin in serum and erythrocytes were measured on a regular basis throughout the study period employing the monoclonal IRMA "Micromedic" kit. RESULTS: Serum iron levels were significantly lower in first degree chronic renal failure (CRF) patients (11.07 +/- 1.54 mmol/l) as compared with the controls (P < 0.01) whereas serum (91.10 +/- 4.00 ng/ml) and erythrocyte (0.83 +/- 0.08 ng/g Hb) ferritin levels were within normal limits. There was a moderately positive correlation (r = 0.37) between serum and erythrocyte ferritin levels. In second and third degree CRF patients the serum iron (7.82 +/- 0.72 mmol/l), serum ferritin (77.60 +/- 3.24 ng/ml) and erythrocyte (0.71 +/- 0.06 ng/g Hb) ferritin levels were significantly lower as compared with those of first degree CRF patients. In the hemodialysis patients erythrocyte ferritin levels showed a tendency towards increase as compared with those in second and third degree CRF patients (P < 0.05). There was a moderately negative correlation (r = 0.44) between serum iron and erythrocyte ferritin levels in this group of patients. CONCLUSIONS: 1. Erythrocyte ferritin levels can be useful in the complex diagnostic assessment of the anemic syndrome in CRF patients. 2. Erythrocyte ferritin is a reliable indicator of iron overload in hemodialysis patients. 3. Erythrocyte ferritin gives no advantage in the evaluation of iron metabolism of medically treated CRF patients. PMID- 10371804 TI - Roentgenographic study of ethosuximide-induced functional changes in rat gastrointestinal tract and their modulation by Nivalin in a chronic experiment. AB - Ethosuximide, a typical antiabsence drug, causes gastrointestinal complaints in the drug-treated patients. In the present study we investigated in an experimental model the functional disturbances occurring in rat gastrointestinal tract (GIT) after a 100-day chronic administration of ethosuximide. Contrast radiographic study of rat gastrointestinal tract was used. The mechanical activity of GIT smooth muscle (SM) preparations was measured using an in vitro isometric technique. The 100-day ethosuximide treatment induced atonia, disturbed peristalsis, and lead to a delay of the contrast material evacuation from the gastrointestinal tract combined with a reduction of the acetylcholine effect on the contractions of GIT smooth muscles in ethosuximide-treated rats. Nivalin cannot eliminate the ethosuximide-induced disturbances in the gastrointestinal tract after a 100-day administration of ethosuximide to the rats. It is thus assumed that Nivalin is not capable of compensating the inhibiting effect of ethosuximide. The authors assume that desensitization of acetylcholine receptors may occur and this reduces the effect of Nivalin action. PMID- 10371805 TI - Family history and some other factors in primary open angle glaucoma. AB - OBJECTIVE: To study the association of a positive family history of glaucoma with age, systemic hypertension, diabetes and refraction in patients with primary open angle glaucoma (POAG). METHODS: Two hundred and five POAG patients were entered into the study. They were allocated to two groups depending on whether or not they had a positive family history of glaucoma: group I included patients with family members with glaucoma and group II--those without family members with glaucoma. Age, sex, refractive error, systemic hypertension and diabetes were analysed as factors in the groups. RESULTS: Sixty seven patients (mean age, 62.6 +/- 1.2 years, SD) had a positive family history of glaucoma. Their mean age was significantly lower (P < < 0.001, F = 17.96) compared with the mean age (68.6 +/- 0.8 years) of the group with negative family history of glaucoma. The frequency of positive history of glaucoma decreased with age. No difference were found with respect to sex, systemic hypertension and refraction between the two groups. There was a prevalence of diabetes in the group with negative family history of glaucoma. CONCLUSIONS: The results of the study suggest that there is no association of family history of glaucoma with sex, systemic hypertension, diabetes and refraction. They show that the disease has an earlier onset in cases with positive family history of glaucoma. PMID- 10371806 TI - Evaluation of the Bulgarian medicine in the ninth century. AB - One of the questions that has not been adequately addressed up to now concerns the level of the official Bulgarian medicine in the ninth and tenth centuries. A reliable assessment of the medicine at that time is possible to obtain by comparing it with the medicine in other countries of the Christian World. The author chooses the Byzantine medicine which is held by the general view to be the culmination of the Medieval medicine in the Christian world. The comparisons between the Bulgarian and Byzantine medicine allows us to make the assertion that the medicine in Bulgaria was far from being at a lower level than that of Bizantium. This fact is worth the attention of the investigators and requires additional research. PMID- 10371807 TI - On the official medical doctrine in Bulgaria after establishment of Christianity as a state religion. AB - The author has been studying the problem of Bulgarian national diet tradition in the ninth and tenth centuries for thirty years. In the course of the research, it was necessary to elucidate what medical knowledge the Bulgarians at that time possessed. The analysis revealed an orderly medical doctrine functioning immediately after the conversion of Bulgaria to Christianity and meeting the requirements of the new faith. As early as the tenth century, converting other peoples to Christianity (the Slavonic tribes primarily), Bulgarians imposed their medical doctrine as part of the Christian propaganda among the newly converted communities. This is one of the contributions of the Bulgarian medieval civilization to the world of that time, a contribution that is not well known and therefore not assessed on its merits. PMID- 10371808 TI - [The epidemiology of musculoskeletal changes due to biomechanical overload of the spine in the manual lifting of patients]. AB - The painful lumbosacral symptoms associated with manual lifting by nursing staff constitutes an increasingly important problem in Occupational Health. This category of workers is in fact particularly exposed to risk situations involving the lumbar region of the spine, due especially to the extreme variability of work on the shift, the nature of what is lifted and not always sufficient knowledge and proper performance of the movements. On the basis of a review of a series of studies made on this topic, it was possible to assess the incidence and prevalence of low back pain in selected populations and identify the postural risk to which nursing staff are exposed at work, stressing also the importance of a correct knowledge of manual lifting techniques, frequency and mode of performing them, and the psychological aspects (perception of the intensity of tasks and osteoarticular strain). It was also observed that the data on painful lumbosacral symptoms were underestimated due to the type of epidemiological investigations carried out (mostly cross-sectional) and that comparison of the data proposed was often difficult due to the different criteria of evaluation used to classify the painful symptoms and also to the various epidemiological parameters used (incidence, cumulative incidence, prevalence). PMID- 10371809 TI - [Instrumental studies in laboratories dedicated to the examination of disk overload in health workers]. AB - The main factors to be considered when applying movement analysis techniques to complex situations are analysed. In particular, when the mechanical interactions of the operator with an object to be handled are not easily measurable, as in the case of lifting a patient in a nursery, an approach in which the ground reaction forces are measured through a force platform is more convenient. In these situations the posture and the movement of the subject, that are also difficult to track, do not need to be analysed. The only information needed is the position and orientation of the intervertebral disk and some estimation of the anthropometric parameters of the part of the body which is below the vertebral level of interest. This can be achieved by a relatively simple approach and a few reasonable assumptions. In addition, the method can enable us to analyse forces and moments in 3-D. The results presented refer to several kinds of operations that are usually performed with hospital patients. They include bed to wheelchair transfer, assistance with standing up from a chair, repositioning the patient in bed. It is shown that the compression loads (1810 N to 5110 N) are comparable to those already reported in the literature. In addition, the transversal components (anterior (260 N-707 N) and lateral (5 N-200 N) shear) are of considerable interest. It is also shown how the use of appropriate ergonomical devices to help patient lifting and handling can considerably reduce the loads on the spine: in fact the compression loads are between 991 N and 1644 N. PMID- 10371810 TI - [The occupational risks and pathologies due to the manual lifting of patients in Italian legislation]. AB - Current Italian legislation establishes all necessary details of intervention procedures aimed at elimination or substantial reduction of risk due to manual lifting of patients. Chapter V of Law No 626/94, which was a thoroughly innovative step in the field of safety, hygiene and prevention at the workplace, in 3 articles and one annex incorporated Community Directive 269/90 into Italian legislation. The model to approach manual load handling, whose definition can in all respects be transferred to health facilities where load means patients who are not self-sufficient in movement, sets out a precise strategy of actions. The first action is automation of the "production process", which is inapplicable in the health care sector; the second action is to provide "aids", which is perfectly applicable to the health care sector. A further action foreseen by the regulations that should be taken into consideration in addressing the problem of manual patient handling from the point of view of prevention is found in the Law (Presidential Decree) of 14/1/97 concerning criteria of accreditation of health facilities, which also refers to the regulations contained in Law 626/94. As regards the insurance coverage of acute or chronic impairment due to manual handling of loads/patients, these aspects are still not contemplated in any regulations. This contradiction became even more evident with the introduction of Law No 626/94 where in Chapter V reference is made to "dorsal-lumbar lesions". Although these lesions are recognized legally in the acute form (accidents), there are also degenerative diseases with chronic development, and the latter are not given any protection apart from the theoretical recognition contained in the Constitutional Court Sentence 179/88. It is to be hoped that the review process of the list of compensable occupational diseases will address this problem, too, so that it can be solved as soon as possible. PMID- 10371811 TI - [The environmental characteristics of patients' rooms and their interactions with the lifting of patients]. AB - The Law (Presidential Decree) of 14/1/97 requested the regions of Italy to define and develop procedures for the accreditation of their health facilities. Since accreditation promotes the quality of the system, the support of ergonomics can be fundamental. Ergonomics, whose quality objective is harmony of the environment man-object relationship, is in a position to make a major contribution to the complete application of the procedures for quality certification (ISO regulation 9000) and of the regulations regarding mandatory safety regulations (Law 626/94). The present paper analyzes the tasks of health workers vis-a-vis patients, highlighting the improvement or worsening effect of environmental features which not only have a direct effect on movements and general activity but also have an indirect effect on psychological and behavioural conditions. The first requirement that should be taken into consideration in environments that are also used by patients is that everything that can facilitate or make their walking and access to facilities self-sufficient will certainly make the healthcare workers' tasks easier, which can then be limited to lifting required for treatment and washing. The ward, especially, is an area that is extremely important from the point of view of design because it is where the patient does practically everything and where the staff, too, are mainly engaged in hygiene/treatment tasks, involving both patients and their relatives. The bed must be able to be easily used by potentially disabled patients who have difficulty in walking and moving. The beds should therefore be equipped with or have close by handles and rails that assist the patient to lie down, get up and move from the bed; the bed should also have a manual or electrical device to adjust the height (from 40 to 70 cm) and have four wheels (that can each be blocked) to make it easier to move. The supporting legs should occupy as little space as possible and should not interfere with wheelchairs or other walking aids or with the movements of the staff. The head and foot-boards should have parts that can be used as grips. The space between both sides of the beds should be at least 90 cm and the space between headboard and wall 120 cm; bedside tables should be no deeper than 30 cm and be fitted with wheels. On the question of space distribution design, another important requirement is flexibility. Adequate flexibility of space distribution together with adaptable technological systems mean that the spaces and equipment of the ward can be easily adapted to the great variety of needs. PMID- 10371812 TI - [The assessment of exposure to and the activity of the manual lifting of patients in wards: methods, procedures, the exposure index (MAPO) and classification criteria. Movimientazione e Assistenza Pazienti Ospedalizzati (Lifting and Assistance to Hospitalized Patients)]. AB - Since a method for quantifying exposure to patient handling in hospital wards is lacking, the authors describe and propose a model for identifying the main risk factors in this type of occupational exposure: presence of disabled patients, staff engaged on manual handling of patients, structure of the working environment, equipment and aids for moving patients, training of workers according to the specific risk. For each factor a procedure for identification and assessment is proposed that is easily applicable in practice. The authors also propose a formula for the calculation of a condensed exposure index (MAPO Index), which brings together the various factors. The exposure index, which requires further, detailed study and validation, makes it possible, in practice, to plan the preventive and health measures according to a specific order of priority, thus complying with the requirements of Chapter V of Law 626/94. From a practical point of view, in the present state of knowledge, it can be stated that for MAPO Index values between 0 and 1.5, risk is deemed negligible, average for values between 1.51 and 5, and high for values exceeding 5. PMID- 10371814 TI - [The assessment of the risk due to the manual lifting of patients: the initial descriptive and analytical results on exposure levels]. AB - The paper reports the results of risk evaluation of patient lifting or moving obtained from a multicentre study on 216 wards, for both acute hospital patients and in geriatric residences. In all situations the exposure to patient lifting was assessed using a concise index (MAPO). Analysis of the results showed that only 9% of the workers could be considered as exposed to negligible risk (MAPO Index = 0-1.5); of these 95.7% worked in hospital wards and only 4.3% in geriatric wards. A further confirmation of the higher level of exposure of workers in long-term hospitalization was that 42.3% were exposed to elevated levels (MAPO Index > 5) compared with 27.7% observed in hospital ward workers. The mean values of the exposure index were 6.8 for hospital wards and 9.64 for geriatric residences and, although much higher in the latter, both categories showed high exposure. In the orthopaedic departments of the hospitals the values were higher than in the geriatric wards (MAPO Index = 10.1); medical and surgical departments showed values similar to the mean values observed in the geriatric wards. These high values were due to: severe shortage of equipment life lifting devices (95.5%) and minor aids (99.5%), partial inadequacy of the working environment (69.2%), poor training and information (96.1% lacking); only the supply of wheelchairs was adequate (65.8%). All of which points to an almost generalized non-observance of the regulations listed under Chapter V of Law No. 626/94. However, the proposed method of evaluation allows anyone who has to carry out prevention and improvement measures to identify priority criteria specifically aimed at the individual factors taken into consideration. By simulating an intervention for improvement aimed at equipment and training, 96% of the wards would be included in the negligible exposure class (MAPO Index 0 1.5). PMID- 10371813 TI - [Clinical studies in health workers employed in the manual lifting of patients: methods for the examination of spinal lesions]. AB - To enable different research groups to make a standardized collection of clinical data on alterations of the lumbar region of the spine, protocols were used for the collection and classification of data that were proposed and thoroughly validated by the authors. The protocols include a clinical/functional examination of the spine, checking for positive anamnestic threshold, for pain on pressure/palpation of the spiny apophyses and paravertebral muscles, for painful movements, in order to classify 1st, 2nd and 3rd grade functional spondylarthropathy (for different regions of the spine). An ad hoc questionnaire was also prepared for the quantitative and qualitative study of true acute low back pain and the ingravescent low back pain controlled at the onset pharmacologically. The results of this questionnaire make it possible to calculate the incidence of acute low back pain (true and pharmacologically controlled). PMID- 10371815 TI - [Initial epidemiological data on the clinical effects in health workers employed in the manual lifting of patients in wards]. AB - An investigation was carried out by teams from various centres coordinated by the EPM (Ergonomics of Posture and Movement) Research Unit on 54 different hospitals in various regions of northern and central Italy. The teams examined a total of 3341 health workers whose job involved manual handling of patients (553 male and 2788 females, 1568 working in hospitals and 1773 in geriatric residences). Numerous meetings were held to ensure that the methods of assessing the exposure indexes and spinal impairment were identical in the various teams. The final data were processed centrally at the EPM Research Unit. The sample analyzed may be considered as representative of the situation in hospitals in Italy, at least for northern and central Italy. The mean age was 36 years, mean length of service in the department 6 years and mean length of job duration not exceeding 10 years; staff turnover was high. Physical examination revealed that 8.4% of the workers had had at least one episode of acute low back pain in the previous 12 months: i.e., 4 times the values of the reference groups. Also in the case of clinical functional spondyloarthropathies of the lumbosacral spine, in the females there was a significantly higher prevalence than in the reference groups. All disorders were more severe in sectors more at risk, i.e., old peoples homes, rehabilitation centres, orthopaedic and surgical departments, and in any case higher in old peoples homes and geriatric residences. The initial data concerning the ratio between presence of spinal disease and risk index were also positive. PMID- 10371816 TI - [Acute lumbago due to the manual lifting of patients in wards: prevalence and incidence data]. AB - The aim of the study was to measure the occurrence (prevalence and incidence) of episodes of acute low back pain (definite effect) in a wide sample of health workers assisting disabled patients. A questionnaire was used for the study both of true acute low back pain and of episodes of ingravescent low back pain controlled pharmacologically at the onset. The questionnaire identified overall acute and pharmacologically controlled episodes occurring in the previous 12 months, both in the course of work and over the whole life of the subject. Appropriately trained operators administered the questionnaire to 551 subjects; 481 valid answer cards were obtained from 372 females and 109 males working in medical, orthopaedic and geriatric departments. 75.4% of the sample had high exposure index levels for patient lifting. The prevalence of true acute low back pain was 9% in males and 11% in females referred to the previous 12 months. Taking acute true and pharmacologically controlled low back pain together the prevalences rose to 13.8% for males and 26.9% in females. Data from the reference populations showed that acute low back pain did not exceed 3% on average in the previous year. Since work seniority in the hospital wards was known, the incidences were calculated, giving 7.9% in females and 5.29% in males for acute low back pain, and 19% in females and 3.49% in males for pharmacologically controlled low back pain. Considering the number of episodes in 100 workers/year, acute low back pain alone reached prevalences of 13-14%. This therefore appears to confirm the positive ratio between episodes of low back pain and duties involving assistance to disabled patients. PMID- 10371818 TI - [The application of a synthetic index of exposure in the manual lifting of patients: the initial validation experiences]. AB - Via a multicentre study coordinated by the EPM research group carried out in 216 wards in a total of 56 hospitals, old peoples homes and geriatric departments, it was possible to quantify exposure to patient handling (classified in 4 classes: 0 1.5 negligible, 1.51-5 slight to average, 5.01-10 average to high, > 10 elevated), and at the same time identify the damage to the lumbosacral spine thus caused. Both assessment of exposure and identification of health impairment were carried out using homogeneous methods. Subjects with work seniority in the job of less than 6 months and subjects who had been transferred because of back trouble were excluded from the study. It was therefore possible to carry out two types of study to assess the association between exposure and impairment. In study A, covering 3021 subjects, an analysis was performed of the association between exposure index, positive response to the anamnestic threshold for lumbosacral disorders and acute low back pain using the method of logistic analysis to obtain the prevalence odds ratios. In study B, covering 418 subjects, the analysis of association was performed on the incidence rates of episodes of acute low back pain and pharmacologically controlled acute low back pain, assuming that exposure in the wards had remained constant. The technique used was Poisson regression, thereby calculating the relative incidence rate ratios. Both for PORs and IRRs the reference group consisted of the exposure class judged as negligible (exposure index 0-1.5). The results showed that the PORs calculated for positive lumbar threshold were significant for increasing exposure classes with a positive trend for the second and third exposure class but not for the last, presumably due to a healthy worker selection effect. Neither in Study A nor in Study B were any associations observed between exposure and acute low back pain occurring in the previous 12 months: this may be due to the fact that the impairment indicator does not appear to be appropriate in terms of latency period. A different result was obtained in Study B which showed a good association between exposure and incidence rates of episodes of acute low back pain and pharmacologically controlled acute low back pain according to department. The IRRs showed a positive trend both for acute episodes (IRR: 1.932, 2.439, 2.847) and for acute plus pharmacologically controlled acute episodes (1.798, 1.830, 4.523). On the basis of these results, even with the caution required for the reasons explained in the text, it seems to be possible to identify three grades of exposure which correspond to increasing probability of impairment of the lumbosacral region of the spine: the first where risk seems negligible corresponds to an exposure level between 0 and 1.5. The second, where the episodes of low back pain may occur with an incidence 3.8 times greater, corresponds to an exposure level between 1.51 and 5, and the third corresponds to exposure levels exceeding 5, where the episodes of low back pain may occur with an incidence up to 5.6 times greater than expected. PMID- 10371817 TI - [Worker turnover because of spinal pathologies in the wards and outpatient clinics of the hospitals of Leno and Manerbo, Brescia]. AB - A total of 306 employees of the hospitals of the Leno/Manerbio health area underwent clinical and anamnestic examination in order to ascertain the existence of degenerative diseases of the spine associated with "manual handling of loads" risk. The prevalences obtained for positive anamnestic threshold concerning the lumbosacral spine, the trend of total acute low back pain and of low back pain in the last year showed lower values compared to the entire national group and in any case lower or only slightly above the values for the reference group of non exposed subjects. Therefore, in order to assess the real prevalences of disorders due to incorrect load handling in hospital environments, it is important to assess the presence of associated disorders of the spine also and especially in outpatients departments. In fact, unsuitable or unfit staff had recently been transferred from the wards to outpatients departments. 56 workers from outpatients departments underwent physical-anamnestic examination: 16 workers (4.5% of the entire group under study) from average-to-high risk wards were identified as suffering from degenerative disorders of the lumbosacral spine. Therefore the prevalence of unfit subjects from hospital wards, cancelling the effect of the turnover factor on outpatients departments, led to an almost twofold total frequency, which rose from 6.9% to 11.4%. PMID- 10371819 TI - [The assessment of exposure to the risk of the manual lifting of patients and the results of clinical studies in the hospitals of Bolzano and Bressanone]. AB - A study was made in two hospitals in the province of Bolzano in order to ascertain the possible relationship between risk due to manual lifting of patients and occurrence of back disorders in healthcare workers. Risk analysis was carried out in 16 wards, with 207 workers assessed by means of clinical/anamnestic check-up of the spine. In the majority of the wards analysed the index of exposure to patient lifting was average or high due mainly to the lack of equipment, unsatisfactory structures and furnishings, and organizational factors. This was particularly evident in the Bressanone hospital where there were fewer workers to each department. As regards the clinical assessment of the spine, from the overall analysis of the results it was seen that in the hospital in Bressanone the prevalence of degenerative disorders of the lumbosacral region of the spine was 3.9% and in the Bolzano hospital it was 5.7%. Similar differences were observed in the prevalences of episodes of acute low back pain reported in the previous 12 months: Bressanone 7.7%, Bolzano 3.8%. In both hospitals 3rd grade SAP was diagnosed in about 5% of the subjects examined and globally clinical/functional alterations of the lumbar spine were observed in 20% of the Bressanone workers and in 27.2% of the Bolzano workers. Management of the cases judged unfit for manual lifting of patients was particularly difficult because of the high percentage of such cases (8-9%). On the basis of these results, the administrations of both hospitals undertook a series of short-, medium- and long-term interventions in order to reduce the risk for the spine. PMID- 10371820 TI - [The assessment of exposure to the risk of the manual lifting of patients and the results of a clinical study in the rest homes of the Mantua area]. AB - At the request of the Geriatric Residences Management Committees, the Prevention and Safety at the Workplace Service of Mantua Hospital coordinated the practical application of Chapter V of Law 626/94 with special reference to patient handling. Assessment of exposure and clinical identification of disorders and impairment of the lumbar spine carried out using the methods proposed by the EPM Research Centre involved 15 residences and a total of 31 departments analyzed and 435 workers submitted to health surveillance by the respective certified occupational physicians. With the exception of one department, the exposure levels were found to be medium-high (MAPO Index > 1.51) considering the number of disabled patients, the significant lack of lifting equipment and the lack of training for this risk factor. Acute low back pain reported in the previous 12 months revealed a prevalence of 10%, more than 4 times that of non-exposed workers. The prevalence of functional spondylarthropathies also showed high values (16%), as also the number of degenerative diseases of the lumbosacral spine (16%). 11% of the males and 17% of the females were judged unfit for patient handling, which created problems of job reallocation. By concentrating the results at the Prevention and Safety at the Workplace Service it was possible to plan interventions for improvement, using as referents for the various residences a working group where the presence of a qualified physiotherapist proved to be particularly useful. Coordination of the certified occupational physicians also improved the quality of health surveillance. PMID- 10371821 TI - [The assessment of exposure to the risk of the manual lifting of patients. The results of a clinical study in a geriatric institute of Milan]. AB - In a geriatric hospital in Milan, with over 1000 beds and 750 healthcare workers (with different skills and training), by means of an accident register, during 1997 an 11% incidence of acute low back pain was recorded, with a mean of 19 days absence from work. For this reason, special attention was given to the assessment of manual patient handling risk and to the consequent effects on the lumbar spine using, for the purpose, the method proposed by the EPM Research Unit. Of the 10 wards analysed none showed a negligible exposure index (MAPO Index < 1.5), only one was in the medium exposure class (MAPO Index = 3.05) and the other 9 wards had high exposure (MAPO Index 5.63-24). A group of 59 men (mean age 35.6 years and mean job seniority 9 years) and 179 women (mean age 36.7 years and mean job seniority 10 years) were examined. The most significant result of the physical examination was the high prevalence of subjects who reported at least one episode of acute low back pain in the previous 12 months (16). This was double the prevalence for the country as a whole (8.3%) and 7 times greater compared to the group not exposed to manual load handling risk (2.3%). The prevalence of degenerative disorders of the lumbar spine in the group under study (4.2%) was also analysed, and is of particular interest if associated with the difficulties in completing the diagnostic process. Lastly, 6.3% of the subjects examined were judged unfit (temporarily of permanently) for work involving manual handling of patients. PMID- 10371822 TI - [The manual lifting of patients: the results of an assessment of exposure to the specific risk and of a clinical study in a rehabilitative institute]. AB - An investigation was carried out in a rehabilitation centre in the Province of Milan in order to quantify exposure to patient handling and assess impairment of the spine induced by this risk factor. Altogether 5 departments were analyzed, with different rehabilitation aims (cardiological, respiratory and neurological); 97 of the 104 exposed staff underwent physical examination by an occupational health physician. Both exposure assessment and identification of impairment were performed using the methods proposed by the EPM Research Group. The exposure indices (MAPO Index) were high in 4 departments (5.6, 6.75, 8.81 and 13.8) and only one department had an index of 2.95 which is considered low-medium. The prevalences of impairment of the lumbar region of the spine were also higher in this group of workers compared to those found in other hospitals. In particular, the disorders of the lumbar spine reported were 45.4% in the males and 62.6% in the females. Of these 13% showed a third grade SAP. About 11% of the workers who underwent physical examination reported episodes of acute low back pain in the previous 12 months with a frequency 4 times higher than that found in group of unexposed workers. Analysis of the rehabilitation centre drew attention to the need for improvement measures that took into account the particular nature of the care services involved as in some cases it is not always possible to use equipment and aids to assist certain manual handling. In these cases the solutions involve the organization aspects especially. The results of this study were used by the rehabilitation centre management in order to plan the interventions for improvement. On the whole it was shown that analysis of this risk factor and health surveillance of exposed workers in such particular structures as rehabilitation centres is of considerable importance. PMID- 10371823 TI - [Exposure to the risk of the manual lifting of patients and the results of a clinical study in 4 hospital establishments of northern Italy]. AB - The paper reports the results of a study carried out in four hospitals in northern Italy to assess exposure to patient handling and the consequent risks for the lumbar region of the spine. The methods proposed by the EPM Research Unit were used. Altogether, in the four hospitals there were 148 wards and 5596 staff. Of these, 34 wards and 510 workers were examined. The results of exposure assessment showed that the obstetrics departments had negligible risk (MAPO Index 0-1.5), urology and general surgery departments an intermediate risk (MAPO Index 1.51-5.0), while the departments of medicine, orthopaedics, neurology and rehabilitation had high risks (MAPO Index > 5.0). Of the 510 workers who underwent physical and anamnestic assessment for spinal disorders, 44% worked in the departments of medicine, which are known to have a high risk The prevalence of subjects who reported episodes of acute low back pain in the last 12 months (11.4%) was 5 times that of a group of workers not exposed to manual load handling (2.3%). Analysis of the same disorder referred to each job showed higher prevalences in the non-nursing staff (technical 25%, general 27%). The frequency of degenerative disorders of the lumbar spine was slightly lower than the figure for the country as a whole (6.7%). It is clear that measures for improvement of the environment are required aimed particularly at installing aids and staff training (for the specific risk factor), also so as to better manage the reallocation of workers judged unfit for patient handling who, in the group under study, were 5%. PMID- 10371824 TI - [Initial data on shoulder pathology in workers exposed to the risk of lifting patients in different rest homes in Trento Province]. AB - A total of 829 subjects (females: 364 aged < 36 years and 360 aged > 35 years; males: 52 aged < 36 years and 53 aged > 35 years) employed on assistance and handling of patients in 42 geriatric residences in the Province of Trento underwent physical and anamnestic examination using a standardized method proposed by the EPM Research Unit for identification of shoulder disorders. The subjects who had reported the existence of pain lasting at least one week in the previous 12 months and occurring at least once a month and not due to trauma of the scapulo-humeral joint were defined as cases with positive history. The shoulder disorders were correlated to the degree of exposure to risk due to handling of patients and quantified with a MAPO Index. The prevalences of disorders of the scapulo-humeral joint in the group under study were slightly higher than those of the workers not exposed to patient handling, however the difference was significant only in females aged over 35 years. The correlation with work seniority was positive. The frequency of scapulo-humeral periarthritis (current and past) in the subjects under study was high (5%); in addition a greater frequency of disorders was observed in subjects with higher degrees of exposure to patient handling. The above observations require further study and confirmation but do not exclude the possibility that the risk involved in handling patients can have effects not only on the dorso-lumbar region but also on other joints, such as in this case the scapulo-humeral joint. PMID- 10371825 TI - [The management of the risk due to the manual lifting of patients in a hospital risk-management program]. AB - Application of Law 626/94 in the health care area is still only partial and in many respects practically in the initial stages. This is due to a number of causes, including the special features of this sector and the extreme variability and diffusion of the risk factors involved. Assessment of risk, especially the newest ones (from manual patient handling to biological agents) involves considerable difficulties, also because the usual methods cannot be applied. The process of assessment and especially the management of risks in this sector is an extremely complex problem, and to address it requires the creation of an organized structure within the hospital consisting of a general manager, or a person delegated by the same, the medical director, the administrative director, the certified occupational physician, the director of prevention and protection. This management team must be flexible in character, draw up strategic plans according to priority criteria, periodically assess the state of advancement of the plans that will be carried out in phases. It is also necessary to ensure the active cooperation of worker safety representatives and of other experts who may be involved in the various topics addressed from time to time. The authors propose a method used in a three-year project, still under way, for the assessment of risk due to manual patient handling in a major Italian hospital; the data obtained from an assessment made in 58 wards were the following: about 60% of the wards showed a medium-high exposure level, 24% negligible exposure levels and 13.8% zero exposure due to the absence of disabled patients. On the basis of the initial data obtained from health surveillance programmes on a group of 431 exposed subjects, about 10% were judged fit with limitations due to spinal disorders. The main problems that have arisen (management, organisation, training, transfer of workers with limited job fitness, accreditation) are discussed. The method proposed for management of risk due to manual handling of patients is considered suitable for other risk factors, within the framework of a more far reaching programme for application of Law 626/94, as a means of permanent management of risk in a particularly complex working environment such as that of a hospital. PMID- 10371826 TI - [The experience of managing the risk due to the manual lifting of patients in the hospital reality of Local Health Screening Unit Enterprise 18, the Veneto region]. AB - The possibility of accidents and diseases connected with manual handling of patients can be considered as one of the major risks in hospital environments for nursing, non-nursing and technical staff. Health surveillance of exposed workers has revealed numerous cases of back disorders with consequent temporary or permanent limitations of working activity. This involves not only heavy costs for the hospital but also a reduction in the quality of the assistance due to the difficulty in replacing or increasing staff. For these reasons the hospitals of the Local Health and Social Services Unit 18 of the Regione Veneto drew up a plan of interventions in order to address this problem and reduce exposure to manual handling of patients. Details of the costs borne by the hospital due to this risk factor are reported; after the implementation of the improvement plan (health surveillance, introduction of aids, training of staff) it is shown how a reduction in costs was achieved, thus demonstrating the efficacy of the measures taken (reduction of 39.2% in days of staff absence due to back disorders). PMID- 10371827 TI - [A training experience for hospital workers employed in the manual lifting of loads (Bressanone Hospital): its contents and the verification of efficacy]. AB - A check-up of the efficacy of training in the specific field of disabled patient handling was made in the Bressanone Hospital. Some members of the staff had been previously trained in this subject and afterwards they organized courses for all the staff engaged in patient assistance. Before and after this course a series of parameters were recorded on appropriate cards, that enabled a comparison to be made of the various positions assumed by the workers during patient handling, with a concise assessment made for each manoeuvre. Altogether 80 manoeuvres were studied in the medical departments, 40 before and 40 six months after the training course. The results obtained were not up to expectations even though on the whole the number of manoeuvres judged totally incorrect was reduced considerably from 43% to 15%. This was due to a lack of coordination in the action plan drawn up for improvement of this risk factor and especially to failure to purchase equipment (for raising patients, minor aids and wheelchairs). It is stressed that training of health care staff on the topic of risk factors present in hospital environments must be considered as a part of hospital management strategy and therefore developed according to well defined programmes and methods. PMID- 10371828 TI - [The application of Title V of Legislative Decree 626/94 to the hospital care environment: a new model of approach to the prevention and safety in work environments service]. AB - The paper reports the results of a study carried out by a Prevention and Safety of Working Environments Service in checking the application of Law 626/94, especially Chapter V, in hospital environments. Via the study it was possible to identify the risks due to manual handling of patients and therefore to manage compliance with the requirements and obligations of the said Law, as regards health surveillance of exposed subjects (programme to ascertain specific job fitness) and development of plans aimed at reducing the specific risks. The project proposed by the EPM Research Unit was submitted to the health administrations via the control service (Prevention and Safety in Working Environments Service) with the aim of experimenting a new method of assessment of risk due to patient handling. In this context, the control service combined enforcement with a more influential role of safety consultant, thereby playing a more modern role of encouraging employers to undertake preventive measures, alongside the traditional role of safeguarding workers' health. PMID- 10371829 TI - [Assessment criteria in the choice of aids for the lifting of patients]. AB - A fundamental part of the prevention strategies aimed at reducing risk due to manual handling of patients is the use of appropriate aids. This paper defines the basic types of aids for hospital wards: patient lifting devices, aids for hygiene and minor aids; and also proposes a procedure for choice of the type of aid: the procedure uses a specific protocol and also analyzes work organization and the environmental features of the ward. The proposed criteria for choice concern in the first place the fundamental requirements of the equipment: safety for operator and patient, simplicity of use and comfort for the patient. Secondly the basic determinants for choice of the type of aid are the type of disabled patient usually present in the ward and the analysis of the movements made in handling patients. On this basis, for each type of aid, the specific features are defined which direct the choice of supply for the various wards that will be adequate and effective both in reducing risk due to manual handling of patients and in improving the comfort of the patients. PMID- 10371830 TI - [The prevention of risk due to the manual lifting of patients: the psychosocial component]. AB - A high percentage of musculoskeletal disorders in nursing staff with the task of patient lifting is reported in the literature. These disorders are considered to be of multiple etiology and attempts have been made not only to assess the physical load but also to identify the direct or indirect influence of organisational and psychosocial factors so that preventive measures be more appropriate and effective (Law 626/94). In this context, in a recent publication NIOSH recalled that psychosocial factors can alter the relationship between exposure to physical loads and the development or the prognosis of these disorders, and stresses that understanding these relationships is the crucial factor in assessment of exposure that can be addressed with preventive and therapeutic measures. A study was carried out on the staff (113 subjects) of a large hospital in northern Italy with departments recognised by certified occupational physicians as at risk for the musculoskeletal apparatus. In order to quantify the working conditions and the disorders of the spine, a protocol proposed by the Ergonomics of Posture and Movement Research Unit of Milan University was further developed and validated. For assessment of psychosocial factors an Italian version of R. A. Karasek's "Job Content Questionnaire" was drawn up and validated by the Centre for Study and Research of Chronic Degenerative Diseases in Working Environments of Milan University. In addition, Borg's Scale was used for perception of physical load, Kurimori and Kakizaki's Scale for mental fatigue, Kjellberg and Iwanowski's Scale for states of stress, plus the Maslach burnout inventory. Nonparametric statistical analysis was applied to determine the influence of physical and organisational and psychosocial factors on disorders of the musculoskeletal apparatus. The results confirm a good agreement between the objective and subjective assessments of "manual patient handling" risk (Kendall p from 0.26 to 0.37, p > 0.001) and the significant relationships between psychosocial factors and disorders of the lumbar region of the spine: past history of episodes of acute low back pain is associated with limited possibility of making independent decisions (U-Mann Whitney z = 2.81, p 0.004) and job insecurity (U-Mann Whitney z = -2.36, p 0.01); episodes of acute low back pain above the threshold in the previous year were associated with low discretionary powers at work. The results will be discussed on the basis of the possible prevention measures. PMID- 10371831 TI - [The management of cases of "conditional" work fitness due to spinal pathologies in health personnel]. AB - On the basis of the trend in the literature, a process was identified, that uses the results of assessment of exposure to manual handling of patients, in order to practically manage, according to defined priorities, the transfer to other jobs of a large number of cases (about 8-10% of the workforce) that were potentially unfit due to back disorders. Aside from the procedures for clinical classification and grading of the severity of the disorders, which have been dealt with elsewhere, general indications are given for the association of cases of disease with assistance to hospitalised patients according to the spinal involvement required, with appropriate evaluation and classification into different degrees of severity. It is recommended to keep the system of association "cases of disease/workplace" dynamic over time so as to guarantee the best possible working conditions practically and technically achievable, especially for these subjects. PMID- 10371832 TI - [Primary HIV-infection: infectious mononucleosis-like presentation with treatment options]. AB - Two patients, men aged 39 and 52 years, are described with a mononucleosis-like syndrome, due to a primary HIV infection. Both patient developed dermatological manifestations. One patient presented with an encephalitis with an inversed CD4/CD8 cell ratio in the cerebrospinal fluid. Primary HIV infection is often missed as a clinical diagnosis. The possible preservation of a strong cellular immune response against HIV itself in case of early start of treatment argues for an immediate intervention with a combination of antiretroviral drugs in this syndrome. PMID- 10371833 TI - [Feminization of medicine]. AB - Since 1985 more than 50% of all medical students in the Netherlands are women. Only a tiny fraction of specialists, professors, directors of hospitals or insurance companies are women, however. Feminization of medicine will balance the skewed male medical culture. This will improve health care for women and career planning for women doctors. Encouraging part time work will generate incentives to enhance the continuity of medical care for patients in general. PMID- 10371834 TI - [Retrieved memories of traumatic events in the youth]. AB - A working party of the Royal College of Psychiatrists has issued a report on retrieved recollections of sexual abuse during childhood and drawn up guidelines for the coping with such recollections. In contradiction to what the working party is asserting, amnesia for sexual abuse in the past may not be excluded. The working party's conclusion that false recollections of negative events can be induced in susceptible persons may be endorsed. As the working party also states, extreme caution is to be exercised in coping with retrieved recollections of childhood traumas, in which no reliable distinction can be made between true and false recollections because independent and objective evidence is often lacking. PMID- 10371835 TI - [Use and adverse reactions of local antibiotics and disinfectants on the skin]. AB - The local antibiotics are an important addition to the treatment of moderately severe skin infections. Development of contact allergy and bacterial resistance, however, are some of the adverse reactions that may occur. Fusidic acid and tetracycline are the most suitable products for the treatment of superficial primary infections of the skin caused by Gram-positive cocci and secondary infected dermatoses. In case of insufficient effect mupirocin may be tried. Mupirocin is an effective antibiotic in the treatment of nasal carriage of Staphylococcus aureus in cases of increased risk of infection (haemodialysis, continuous peritoneal dialysis, after thoracic surgery). Besides it is an important local antibiotic in the treatment of meticillin-resistant staphylococci in the nose. In order to prevent development of bacterial resistance its further use has to be restricted. Silver sulfadiazine is a good option for the local treatment of infections by Pseudomonas aeruginosa and other Gram-negative bacteria. Erythromycin and clindamycin are useful local antibiotics for the treatment of mild acne vulgaris. Disinfectants are mainly suitable for the use on the intact skin. PMID- 10371836 TI - [Trauma during pregnancy]. AB - Mortality due to trauma in pregnancy is not very common in the Netherlands. More often a pregnant woman presents herself for examination after trauma. Blunt trauma is more common in the third trimester. Minor trauma also needs good care, with special attention for solutio placentae. Maternal mortality after penetrating trauma is low because of the protection of vital organs by the uterus. With good treatment the mortality in pregnant trauma patients will not be higher than in nonpregnant patients. A rapid and effective resuscitation of the mother will give the foetus the best chance of survival. PMID- 10371837 TI - [Immunology in medical practice. XXII. T-cells and strategies for tolerance induction]. AB - Up to the present, aspecific immunosuppressive medication is still required to prevent rejection of transplanted allogeneic tissues and to treat autoimmune disorders. These drugs are associated with numerous adverse effects. Therefore, alternative forms of immunomodulation are being extensively studied. T-cells play a central role in regulating the immune system. Expanding insight into the cellular mechanisms involved in immune recognition and activation led to the development of new tolerance-inducing strategies. Potential sites for tolerance induction are situated centrally in the thymus as well as in the peripheral T cell population. Explicitly mentioned tolerance strategies are (a) allogeneic bone marrow transplantation and intrathymic tolerance induction (central tolerance induction), (b) peripheral T-cell clonal deletion, induction of anergy and immune deviation (all three are examples of peripheral tolerance induction). PMID- 10371838 TI - [Tissue donation in nursing homes; a survey of the number of potential donors and the knowledge and attitude of nursing home doctors and directors]. AB - OBJECTIVE: To obtain an impression of the tissue donor potential in Dutch nursing homes and of the knowledge and the attitude of nursing home physicians and nursing home directors with regard to tissue donation. Also, to gain insight into the problems associated with activating tissue donation in nursing homes. DESIGN: Descriptive and inventory. METHOD: The donor potential was calculated from data derived from the National nursing home registration system (SIVIS) in 1995. A questionnaire on the knowledge and attitude with regard to organ/tissue donation was sent to 400 randomly chosen nursing home physicians and all 323 nursing home directors. RESULTS: Out of the 10,619 somatic patients deceased in nursing homes in 1995, 2670 (25%) would have been suitable for skin and/or cornea donation. Other patients had comorbidity regarded as a contraindication for donorship or were over 80 years of age. Of the 9771 deceased psychogeriatric patients, virtually none were suitable as tissue donors. The response to the questionnaire was 55% among nursing home physicians and 66% among nursing home directors. Both groups showed inadequate knowledge with regard to tissue donation. Most nursing home physicians (85%) took a neutral position on tissue donation in nursing homes; most directors (88%) would support tissue donation in their nursing home. The two problems that were mentioned most in having a negative influence on tissue donation in nursing homes, were: the lack of knowledge of physicians and the refusal of donation by patients. CONCLUSION: Some 2700 somatic nursing home patients annually would be suitable for tissue donation. Determination of their willingness is necessary. Activating of tissue donation in nursing homes requires solving of the lack of knowledge. PMID- 10371839 TI - [Tracing of patients with familial hypercholesterolemia in the Netherlands]. AB - OBJECTIVE: To inventory the possibilities of tracing relatives of patients with familial hypercholesterolaemia (FH) by means of family tree research and DNA diagnostics. DESIGN: Descriptive. METHOD: Blood from patients with the clinical diagnosis of 'FH' was sent, through one of the lipid outpatient clinics in the country, to the Foundation for Tracing Hereditary Hypercholesterolaemia (StOEH) for DNA examination, to characterize the genetic defect. If a mutation was diagnosed in this index patient, he was invited by telephone by a StOEH staff member to have DNA testing done in relatives (especially those of the first degree). The data were stored in a data base. The analysis concerns the patients approached in 1994-1997, as well as those in whom the serum concentration of LDL cholesterol was also determined in 1993-1995. RESULTS: A total of 3013 persons were approached and examined: 146 index patients and 2867 relatives. The DNA diagnosis of 'FH' was made in 1067 relatives (37.2%), 585 (54.8%) women and 482 (45.2%) men. Of these, 21.2% were younger than 20 years, 37.0% 20-39 years, 26.6% 40-59 years and 15.2% > or = 60 years; 44.1% reported being known with a raised cholesterol level, 29.4% were treated with cholesterol-reducing drugs and 6.1% were suffering from a cardiovascular disease. Of the 990 persons in whom the serum LDL cholesterol level was determined, 325 (32.8%) were carriers of a mutation in the LDL receptor gene. 21.2% Of them had a LDL cholesterol level < P95. In the non-carrier group, 14.6% had a serum LDL cholesterol level > P95. CONCLUSION: Tracing FH patients is feasible in practice and leads to detection of as yet untreated patients. PMID- 10371840 TI - ['There is a need for women doctors to join forces': Dutch Association of Women Physicians, 1933-1998]. AB - After a history of some years, the Dutch Association of Medical Women (VNVA) was founded in 1933. The aim of the Association was to promote the interests of women doctors. At that time about 5% of the physicians were women. Their position on the labour market was not at all equal to that of their male colleagues. In 1949 some VNVA members played an important part in stimulating the recognition of women's interests by publishing on anticonception in this journal. It is still necessary to pay attention to equal opportunities for women doctors. Besides, promotion of a sex-sensitive health care was added to the mission statement. The Association nowadays has more than 2600 members. PMID- 10371841 TI - [Women and part-time employment in the medical profession]. AB - During a conference of this journal the following matters were raised: (a) part time work in medicine, although generally accepted socially for men and women is not precisely advisable for those who want to move up to higher functions; (b) owing to the long duration of medical studies, physicians are late to marry and raise a family; (c) the ambition of medical women, amply present at the medical finals, often proves to extend no further than pregnancy; (d) the career of female doctors shows a kink caused mostly by family duties and inadequacy of day nurseries; (e) the current excessive workload of (senior) medical functionaries reduces many women's ambition to attain such functions. Women who want to study medicine should consider much earlier than is nowadays the case what they want and can become within the limits of their possibilities in the wide field of medicine. Energetic striving for shortening of the training period may result in earlier fulfilment of the wish to have children. PMID- 10371842 TI - [40th Congress of the German Society for Pneumology and 25th Congress of the Austrian Society for Lung Diseases and Tuberculosis. Bad Reichenhall/Salzburg, 17 20 March 1999. Abstracts]. PMID- 10371843 TI - [Influence of the equality of ventricular pressure in the echocardiographic determination of the ejection fraction of the left ventricle]. AB - OBJECTIVES: Limited data have been reported concerning the influence of right ventricular systolic overload on the accuracy of the 2D echocardiographic (echo) determination of left ventricular (LV) ejection fraction (EF). The normal newborn at birth (high pulmonary peak systolic pressure) and just after the fall of pulmonary pressure, represent an in vivo model to study this influence. This study compares the LVEF determined by recommended 2D echocardiographic algorithms with that by 3D echo, in newborns at birth and just after the normal fall of right ventricular systolic pressure. METHODS: 100 echocardiographic studies (50 at 2 to 6 hours after birth--group I; and 50 at 7 to 14 days old--control group) were performed in 82 normal newborns, to determine LVEF by 4 geometric models (cylinder hemiellipsoid; ellipsoid biplane; single plane area length, in 4- and 2 chamber view; biplane method of discs) and visual estimation, using 3D echocardiography as the reference method. RESULTS: In group I, the correlation between 3D echo and cylinder hemiellipsoid was r = 0.62 (SEE = 4.5%); ellipsoid biplane, r = 0.69 (SEE = 4.1%); single plane area length, 4 chambers, r = 0.66 (SEE = 5.1%) and, 2 chambers, r = 0.72 (SEE = 4.0%); biplane method of discs, r = 0.83 (SEE = 3.6%), and, visual estimation, r = 0.78 (SEE = 3.5%). In the control group, the correlation between 3D echo and cylinder hemiellipsoid was r = 0.70 (SEE = 3.4%); ellipsoid biplane, r = 0.63 (SEE = 3.4%); single plane area length, 4 chambers, r = 0.79 (SEE = 3.5%) and, 2 chambers, r = 0.76 (SEE = 4.1%); biplane method of discs, r = 0.90 (SEE = 2.3%), and, visual estimation, r = 0.64 (SEE = 3.9%). IN CONCLUSION: These data suggest that the biplane method of discs and single plane area length using 2-chamber view allows a more accurate LVEF determination when significant right ventricular pressure overload is present. PMID- 10371844 TI - Cellular immune responses to Mycobacterium tuberculosis in a patient with Takayasu's arteritis. AB - INTRODUCTION: Takayasu's arteritis (TA) is a disease of unknown aetiology, characterized histologically by an inflammatory cell infiltrate that affects all layers of the arterial wall. Its association with tuberculosis (TB) was described 50 years ago, based on the presence of Langhan's giant cells and granulomas similar to those found in tuberculous lesions. The presence of TB in patients with TA well as been reported in several studies as well as a positive tuberculous response, but these associations could be fortuitous in countries where TB is endemic. Recent studies have shown that patients with TA have a heightened humoral response to mycobacterial antigens including the 65 kDa fraction, a heat shock protein (HSP) that has also been found to be expressed in the arterial wall of patients with TA. The purpose of this study was to determine lymphoproliferative response and interferon-gamma (IFN-gamma) production by peripheral blood mononuclear cells (PBMC) stimulated by live Mycobacterium tuberculosis (Mtb) H37Rv and a panel of mycobacterial antigens, in the hope of contributing to a better understanding of the cellular immune responses to Tuberculosis in Takayasu's arteritis. MATERIAL AND METHODS: Standard lymphoproliferation tests and IFN-gamma determination (ELISA) were performed in a 47-year old black man who fulfilled criteria for TA and 10 healthy controls, BCG vaccinated, Mantoux positive. The following were used: Mtb H37Rv, Purified Protein Derivative (PPD), purified 30 kDa, recombinant M. bovis BCG 10 kDa, 38 kDa, 65 kDa, 70 kDa, Short Term-Culture Filtrate Proteins (ST-CFP), Mid Term Culture Filtrate Proteins (MT-CFP) obtained from H37Rv and phytohemaglutinin (PHA) as mitogen for positive control. RESULTS: PBMC from the patient with TA when compared to the mean values of the 10 healthy donors showed decreased proliferation in response to all antigens, with the exception of 65 kDa. The TA patient showed a similar pattern of IFN-gamma production to that obtained with control donors, with the exception of higher IFN-gamma production in response to ST-CFP and MT-CFP. CONCLUSIONS: We have shown reactivity of peripheral lymphocytes to HSP 65 kDa and a trend towards higher production of IFN-gamma in response to ST-CFP and MT-CFP in a patient with TA. These facts, together with the already established heightened humoral response, strengthens the association between TB and TA. However, HSP 65 kDa is not specific to TB and we conclude that similar studies using lymphocytes obtained from the arterial wall of TA patients may help to clarify the role of mycobacterial infection in Takayasu's arteritis. PMID- 10371845 TI - [Myocardial infarct without angiographic coronary atherosclerosis: study of a group of patients]. AB - STUDY OBJECTIVES: Clinical characterization and aspects of subsidiary clinical tests in a group of patients with myocardial infarction with no visible angiographic atherosclerotic stenosis. Etiologic identification, therapeutic approach and prognostic assessment of non atherosclerotic myocardial infarction. PATIENTS: We studied patients admitted with myocardial infarction to coronary care unit over a 4 year period and in whom cardiac catheterism did not show atherosclerotic coronary stenosis (17 patients). METHODOLOGY: Retrospective study; Assessment of clinical characteristics, exercise test parameters, echocardiogram, hemodynamics, heart rate variability (HRV) and signal-averaged electrocardiogram (ECG) of this group of patients; Maximal follow-up of 44 months and minimal of 3 months (average: 19.9 +/- 12.7 months) for the occurrence of ischemic cardiac events (recurrent angina, reinfarction or sudden death). RESULTS: Non atherosclerotic coronary infarction was an unusual situation (2% of the totality of the infarction hospitalized during those 4 years--795 cases), occurring mainly among young men, with few vascular risk factors (except smoking), as small infarctions, without a preferential localization and with good evolution in the acute phase (Killip I). In the predischarge exercise test there was no residual ischemia and functional capacity was generally good. In the majority of cases left ventricular systolic function was preserved (82% of the cases). With cardiac catheterism, we observed two cases of "bridging" and four cases of slow contrast progression. In arrhythmic risk stratification with 24 hours ECG, HRV and high resolution ECG, we observed no adverse prognostic markers in the majority of the cases. The clinical observation of the patients and the tests permitted us to establish probable hypotheses for the etiological diagnoses in 10 of the cases (left main anterior descending artery "bridging"--2 cases; slow contrast progression in the coronary vessels--4 cases, severe aortic stenosis--1 case; left valvular mechanic prosthesis--1 case: probable coronary thrombosis with complete reperfusion after thrombolytic therapy--2 cases). The therapeutic approach in the acute phase was the same as that of atherosclerotic infarctions. Secondary prevention was individualized and according to each case etiology, maintaining the antiplatelet agents. In the follow-up there was unstable angina in 3 patients. There were no cases of reinfarction or sudden death. CONCLUSIONS: This study allowed the characterization of the group of non atherosclerotic myocardial infarction as a group of young men with few vascular risk factors, with small infarctions and good prognosis, without adverse arrhythmic risk markers. It also allowed to identify the probable infarction etiology in 10 patients and the secondary individual prevention for each situation. We noted a good prognosis of this situation at two years. PMID- 10371846 TI - [Experimental anatomo-pathologic study in the myocardium of animals with arterial hypertension caused via a nitric oxide synthesis blocker]. AB - INTRODUCTION: Cardiac hypertrophy and reactive (including perivascular) and reparative fibrosis in L-NAME model are similar to that of hypertensive and cardiomyopathic patients. THE AIM: To qualitatively evaluate the reactive (including perivascular) and reparative fibrosis, on days 21 and 35, occurring in hearts submitted to the L-NAME model, using special staining. MATERIAL AND METHODS: We utilized 33 normotensive Wistar rats. L-NAME was administered orally- in a concentration of 75 mg x 100 ml-1 in drinking water. Six rats were submitted during 21 days and an other 15 rats during 35 days. The arterial pressure was obtained on days 12, 20 and 34 using hydraulic plethysmography. On days 21 and 35 during the experiment the animals were anesthetized and submitted to cardiotomy. The hearts were fixed in Bouin fixative during 48 hours and processed using routine methods, emblocked in paraplast and cut in slices 4 to 7 microns thick. The special staining utilized were: Gomori's trichrome (aniline blue) or Masson trichrome, picro-sirius red-polarization, alcian blue technique (pH 0.5 and pH 2.5), periodic acid-Schiff technique (with and without amylases) and Weigert's resorcinol fuchsin solution (with and without oxon). RESULTS: Our results demonstrated an increase in the arterial pressure in animals submitted to the model. On day 21 of submission we observed modest to extensive infarct areas in the right and left ventricular myocardium. On day 35 the wide reparative areas were from old infarct areas. In some cases at day 35 the lesions reached the totality of the right ventricle in several histological slices. The right ventricle was much more affected than the left one. In both groups perivascular fibrosis was observed, nevertheless, on day 21 it was very reduced. DISCUSSION: The degree of direct influence of NO or hypertension produced by NO on hypertension and cardiac lesions during L-NAME model is discussed nowadays. Nevertheless, the fact is that NO deficit has great influence in several cases of cardiac lesions occurring in hypertension and also in cardiomyopathies. PMID- 10371847 TI - [Ischemic cardiopathy in the hypertensive]. AB - Arterial hypertension and coronary heart disease (CHD) are the clinical expression of cardiovascular remodelling, the altered cell population of the cardiovascular system being the result from many biological, psychological and social factors and genetic, enzymatic, humoral and metabolic dysfunction of the individual subject. Both diseases are affected by many common factors and have some similar aspects of the remodeling process, and these are reasons for their simultaneous presence in many patients. The assessment of the total burden of CHD risk in the hypertensive patient and the strategy to modify lifestyle and reduce not only the blood pressure, but also the total burden of risk, is therefore important. PMID- 10371848 TI - [Is the J-curve important in the treatment of the patient with arterial hypertension?]. AB - Stroke morbidity and mortality is related to arterial hypertension. The benefit of arterial hypertension therapy is evident, but the same is not true for coronary artery disease prevention. In recent years, some authors have suggested that as excessive drop in diastolic arterial pressure with therapy could be deleterious. The lack of objective evidence of such an effect required a special study design. It was achieved with the Hot study. After this study, we can probably conclude that the j-curve effect may not be relevant in arterial hypertension therapy. PMID- 10371849 TI - [Therapy of arterial hypertension with angiotensin receptor blockers]. AB - Angiotensin II antagonists block the actions of angiotensin II by occupying the AT1 receptors. With this blockade there is no bradykinin increase, the angiotensin II synthetized by the cardiac chymase is also blocked, and the AT2 receptor is stimulated (antiproliferative effect). In animal experiments, losartan reverses left ventricular hypertrophy, inhibits myocardial fibrosis and diabetic glomerulosclerosis and significantly protects from vascular cerebral diseases. In humans, the efficacy of the angiotensin II antagonists and that of other antihypertensives is similar and is potentiated by the addition of a thiazide. They are very well tolerated and no important adverse reactions are reported. Losartan decreases insulin resistance, has a very favourable hemodynamic and neurohormonal profile in patients with cardiac insuficiency, reverses proteinuria and has a uricosuric effect. Angiotensin II antagonists are a step forward towards the ideal antihypertensive drugs. PMID- 10371850 TI - Accuracy and feasibility of contrast echocardiography for detection of perfusion defects in routine practice. Comparison with wall motion and technetium--99m sestamibi single--photon emission computed tomography. PMID- 10371851 TI - [Hypertriglyceridemias]. AB - Triglyceride atherogenicity is still a matter of controversy for reasons related to their biological and epidemiological characteristics. The authors discuss and review several of these problems, as well as the metabolic relationship of triglycerides and HDL-cholesterol, data indicating a strong influence of triglycerides on cardiovascular risk, mechanisms mediating the pathogenicity of triglycerides, and some questions of drug treatment of hypertriglyceridemias. PMID- 10371852 TI - [The patient with syncope--which are the most useful clinical parameters for defining a diagnostic strategy?]. AB - Syncope is a frequent diagnosis, but the complete definition of its etiology is not uncommonly a difficult process, in which the physician sometimes needs to perform extensive diagnostic work-up, with low yield or inconclusive data. A thorough clinical evaluation allows, nevertheless, a definition of some criteria of great value in the choice of the most productive line of investigation, as well as in assessing a prognosis for the individual patient. PMID- 10371853 TI - [Guidelines for the therapy of heart failure: importance of the treatment individualization]. AB - Heart failure therapy has evolved significantly in recent years, allowing a prolonged survival and better quality of life. However, mortality and morbidity remain high, with enormous costs for the community. The incidence and prevalence of heart failure are increasing in Europe and the United States, predominantly in the elderly. The etiology of heart failure is different nowadays, with about 60% of the patients suffering from coronary artery disease. Modern treatment of heart failure involves general measures common to all patients, regardless of etiology, phase and NYHA class--etiological treatment, correction of precipitating factors, prevention, diet, programmed exercise and angiotensin converting enzyme inhibition. However, the diversity of pathophysiological and etiopathogenic aspects, the symptom intensity and the severity of the disease require individualized therapy, which implies the wise management of a panoply of drugs, mechanical resources and surgery. PMID- 10371854 TI - Prolonged fever in a male patient with a pacemaker. PMID- 10371855 TI - [The College of Cardiology of Coimbra]. PMID- 10371856 TI - About the "Collegium oto-rhino-laryngologicum amicitiae sacrum". PMID- 10371858 TI - [Thyroglossal duct cysts, surgery and histology]. AB - 70 years ago Sistrunck described a specific procedure for the management of thyroglossal duct cysts. However, this surgical procedure is not performed often. In a review on 28 cases, the authors have determined whether the Sistrunck's operation was too extensive in the treatment of thyroglossal duct cysts. 28 surgicals procedures have been performed during five years, 6 Schlange's procedure and 22 Sistrunck's procedure. We have had 17% of complications with only one recurrence, after six months, with Schlange's operation. During the interventions, we have been able to see and feel a duct in only one case. This difficulty in determining the presence of a duct intra-operatively could suggest that there was no duct. So we have undertaken a histological study of all 28 specimens obtained from surgery. Results showed the presence of one or multiple tracts in 72% of cases. Finally, this study show that Sistrunck's procedure is still the best operation for treatment of all cases of thyroglossal duct cysts. All other operations, and particularly Schlange's procedure, are inadequate because they are in contradiction with histological and embryological studies. PMID- 10371857 TI - [Elements of epidemiology and initiation of carcinogenesis in carcinomas of the upper aerodigestive tract. Future therapeutic consequence?]. AB - The study of epidemiology and of the carcinogenesis in epidermoid carcinomas of the upper aerodigestive tract shows that their occurrence is not random. Tobacco abuse plays a major role, especially because of benzopyrene, mutagen of the P53 gene, however it is associated with many other potentiating factors: alcohol, metals, hydrocarbures, virus, food, climate, genetic fragility that create genetic lesions at the origin of carcinogenesis. The latter occurs as "field cancerization" with multiple alterations of the mucosa and general attack of the control systems of the differentiation, growth and cell apoptosis which usually protect the cell against the phenomena of carcinogenesis. The P53 protein gene, retinoid receptors as well as the system of detoxifying glutathion S transferase are modified at the very early stage of these diseases, these abnormalities can be logically related to epidemiological data. These data lead us therefore to imagine complementary specific reverting therapies of induced genetic abnormalities, through the reexpression of non mutated gene encoding P53 protein and the use of retinoid. These various modalities are reported hereafter. PMID- 10371860 TI - [Middle ear ossicular replacement prostheses: cause of failure]. AB - There are various prostheses used for ossiculoplasty in chronic middle ear disease. Plastipores are one of the most commonly used prostheses. We performed ossiculoplasty with plastipores in 237 ear operations in a five years period. The results of the operations were summarized and causes of prosthesis failure were discussed. In 55 (23.2%) patients, tympanic membrane perforation recurred. Cholesteatoma recurrence was encountered in 16 (6.8%) cases. The extrusion of prosthesis was observed in 10 (4.2%) cases. The underlying ear disease was cleaned in 156 (65.8%) patients. An air-bone gap to within 20 dB was achieved in 87 (55.8%) ou of 156 patients. In 69 (44.2%) the gap closure was more than 20 dB. The experience of the surgeon in otologic surgery, severity of the chronic ear disease, status of the eustachian tube, extention of the ossicular chain disruption, type of surgery and characteristics of the prosthesis are critical for ossiculoplasty. PMID- 10371859 TI - [Our experience of carcinoma of the larynx in Togo]. AB - This is a retrospective study of 33 patients with squamous cell carcinoma of the larynx from the end ward at Tokoin Teaching Hospital in Lome. This series is from 1981 and 1995 (1 T1, 6 T2, 15 T3, 11 T4). This is an uncommon cancer. No etiological factors were identified. The high proportion requiring emergency tracheotomy (36.36%) and of T3-T4 cases (78.78%) are explained by the long delay before the first medical visit. Only 13 patients (36.36%) had a total laryngectomy. The prognosis was unfavourable. PMID- 10371861 TI - Temporal bone pathology findings due to drowning. AB - It has been reported that anoxia due to near-drowning or near-suffocation causes brain damage but not inner ear damage. On the other hand, it has been shown that brain death causes both brain damage and inner ear damage. However, studies of temporal bone pathology resulting from sudden death due to drowning are few. We studied temporal bone pathology in six cases of individuals who died of accidents due to drowning. In all temporal bones examined, we found extensive congestion petechiae and haemorrhage in the vessels in the mucosal layers of the middle ear and mastoid air cells, as well as in the vessels around the facial nerve and carotid canal. In the inner ear, there was no abnormality in Corti's organ or the vestibular organs, except in one case who died in the bath. Our findings suggest that petechiae haemorrhage or congestion in the vessels of the mucosal layer and the vessels themselves of the middle ear occurs upon acute death due to drowning. PMID- 10371862 TI - Metastatic thyroid carcinoma of the mandibule. AB - A case of metastatic papillary carcinoma to the mandible is presented. Though relatively rare, metastatic tumours of the mandible should be included in the differential diagnosis of the tumours in the parotid region. For the primary site; being in the cervicofacial region, the thyroid gland must be considered by the head and neck surgeon. PMID- 10371863 TI - The adenocarcinoma of the larynx: in aim of a clinic case. Histologic, immuno histochemistry and electron microscope studies. AB - Adenocarcinoma "not otherwise specified" of the larynx appears to be extremely rare. We report a case of adenocarcinoma "NOS" arising in the larynx of a 72 year old white man. The neoplasia was ALCIAN and PAS negative. Immuno-histochemical studies revealed negative immunoreactivity from NSE, chromogranins and S100; cytokeratin was positive. Electron microscopy revealed electrodense granules of varying sizes, but with no obvious central core surrounded by a limiting membrane. The patient underwent a total laryngectomy and functional neck dissection, with mandibular extension. And he is free of tumor after 4 years postoperatively. PMID- 10371864 TI - Liposarcoma of the hypopharynx. A case report and review of the literature. AB - A new case of well differentiated hypopharyngeal liposarcoma is reported. The author reviews the literature about the clinical and histologic features of these tumors. From the 93 head and neck liposarcomas reported 13 are located in the hypopharynx. The mean age of presentation is 61 years and males are largely predominant. Etiology is still unknown. Tumor size does not seem to affect the prognosis. Usually patients do not present with cervical lymph node metastasis nor distant metastasis. Histologic diagnosis according to Enzinger and Weis's classification could be difficult especially to distinguish between lipoma and well differentiated liposarcoma. The main prognostic factor is histologic grade but early recognition combined with a complete surgical excision can result in a decreased local recurrence rate and high survival rate. Low grade tumors often recur locally but distant metastases are rare. From the 8 well differentiated tumors reviewed 6 presented a local recurrence 2 months to 20 years after surgery but only 1 patient died without disease. High grade tumors are much more aggressive locally and metastasize frequently. Radiotherapy and chemotherapy are proposed in selected cases without evidence to be of value. PMID- 10371865 TI - [Jugular bulb diverticular and facial paralysis]. AB - The authors report a case where a woman presents a right jugular bulb procidence already known and responsible of a perception deafness. Secondarily, a right facial paralysis is appeared progressively and not regressive even with medical treatment. When the computed tomography as shown an intrapetrous diverticular, the facial paralysis treatment was surgical to decompress the facial nerve. The literature study shoes the rarity of this association facial paralysis and jugular bulb procidence (only two cases), more often responsible of deafness, tinnitus, and vertigo. The diagnosis is given by computed tomography. M.R.I. has not still be evaluated. Then the authors insist on the progressive character of the facial paralysis and on the necessity of a surgical treatment. PMID- 10371866 TI - Meningoencephalic herniation into the middle ear. AB - Meningoencephalic herniation into the middle ear (MHME) is a rare condition. It can result from ear surgery, infection, head trauma or can be spontaneous. Diagnosis requires a high degree of clinical suspicion. The presentation may suggest the condition, but sometimes the intraoperative discovery of an occult meningoencephalic herniation may be a frightening situation. Treatment planning must avoid intra-cranial complications. Transmastoid (TM) and middle cranial fossa (MCF) are alternative or complementary approaches, determined by several factors, including the size and the site of the bony defect and the presence or absence of middle ear infection. Three case reports are presented and a review of the literature is performed, to explain some aspects related to MHME, including aetiopathogenesis, clinical presentation, histopathology, diagnosis and treatment. PMID- 10371867 TI - Autofibrin glue compound and its utilization during reconstructive operations on the ear. AB - A new autofibrin glue compound (AFGC) is suggested. The experiment has demonstrated that the inclusion of antibiotics and lysozyme does not influence its adhesive qualities or sterilization with gamma radiation. It has been revealed that the dose to sterilise the compound was 75 Gy. AFGC was used during tympanoplasties in 55 patients for fixation of ossiculoplasties, fascia autotransplant and skin of the external auditory meatus. Morphological and functional results of tympanoplasties turned out to be better than those of the control group in which the glue was not used. PMID- 10371868 TI - [Gastroesophageal reflux in adults]. PMID- 10371869 TI - Nuclear weapons and the law. AB - The history of the International Court of Justice (ICJ) is summarized, with a discussion of some of its earlier Advisory Opinions. The Advisory Opinion on the legality of nuclear arms is considered in the light of the principles of international humanitarian law and a review of nuclear weapons effects. The present government's position on nuclear weapons as outlined in the Strategic Defence Review (which ignores the issue of legality) is examined critically. PMID- 10371870 TI - Detecting the health risks of radiation. AB - Radiation can cause both non-stochastic (cell-killing) effects, leading to burns, epilation, immune system damage and lens opacities, and mutational or stochastic effects due to low dose damage to single cells. If the latter are followed by clone formation or fertilization, the mutants are not recognized by the immune system, and there is no competing cause of death, cancer or leukaemia can result. These effects did not become public knowledge until after the A-bombings of Hiroshima and Nagasaki. Subsequent analysis of the data on A-bomb survivors suggests, contrary to official views, that the immune system has a complex role in the aetiology of cancer and leukaemia, and that the A-bomb survivors were unusually resistant to the harmful effects of the bombings. These findings require the re-evaluation of the effects of low-level radiation, which has increased with the growth of the nuclear industry, both civil and military. PMID- 10371871 TI - Post-invasion change in the trend of complications and outcome of pregnancy in Maternity Hospital Kuwait from 1981 to 1995. AB - To assess the trend in the complications and outcome of pregnancy in Maternity Hospital, Kuwait, a retrospective analysis of yearly hospital statistics books and labour ward records of patients delivering in Maternity Hospital Kuwait was carried out for the period 1981 to 1995. In the post-invasion period there is a significant rise in: primiparity; mothers aged 35 years or older; Kuwaiti mothers; and in multiple pregnancy. The incidence of pregnancy-induced hypertension, pre-eclampsia, and hysterectomy for postpartum haemorrhage also increased. There was a significant increase in spontaneous abortions and low birth weight babies. The incidence of hydatidiform mole has significantly decreased. Still birth rate shows a decreasing trend in the study period. The significant change in the age and parity of the mothers delivering in the post invasion period might partly explain the above changes. However, the effect of environmental pollution, social and psychological stress, and anxiety due to war may have also contributed to an increase in the complications and adverse outcomes of pregnancy. PMID- 10371872 TI - Retrospective revaluation and inhibitory associations: does perceptual learning modulate our perception of the contingencies between events? AB - The reported experiments seek to reconcile the conflict between proposed explanations of retrospective revaluation of contingency judgements in experiments with humans (e.g. Dickinson & Burke, 1996) and those of a perceptual learning phenomenon discovered in rats (Espinet, Iraola, Bennett, & Mackintosh, 1995). The experiments employ a medical diagnosis scenario, in which subjects are asked to judge whether particular symptoms signal the presence or absence of a fictitious disease. In Experiment 1, following alternating exposure to pairs of symptoms, AX and BX, subjects were informed that symptom A was predictive of illness. As a result of this training, subjects rated symptom B as predictive of the absence of the illness. In Experiment 2, a possible mechanism behind this effect was considered. In particular, it was established that pre-exposure in which the AX pair reliably preceded the BX pair produced the same pattern of results as those shown in Experiment 1. In contrast, when the order of pairs was reversed, so that BX reliably preceded AX, the reduction in ratings for symptom B was not obtained. Several extant associative theories are discussed, but none are found to account adequately for this pattern of results. To overcome these inadequacies, a parsimonious theory is suggested that explains all the effects observed in these experiments. PMID- 10371873 TI - Language-specific knowledge and short-term memory in bilingual and non-bilingual children. AB - The sensitivity of children's phonological short-term memory performance to language-specific knowledge was investigated in two experiments. In Experiment 1, monolingual English children, English-French bilingual children, and English children who were learning French as a second language were compared on measures of phonological short-term memory and vocabulary in the two languages. The children's short-term memory performance in each language mirrored their familiarity with English and French, with greater vocabulary knowledge being associated with higher levels of recall of both words and nonwords in that language. This finding was replicated in Experiment 2, in which two groups of children with good knowledge of English and French were examined: native bilingual children who had comparable knowledge of the two languages and non native bilingual children who had a greater knowledge of their native than second language. The findings indicate that phonological short-term memory is not a language-independent system but, rather, functions in a highly language-specific way. PMID- 10371874 TI - Inferring sublexical correspondences from sight vocabulary: evidence from 6- and 7-year-olds. AB - We report an experiment designed to investigate 6-to-7-year-old children's ability to acquire knowledge of sublexical correspondences between print and sound from their reading experience. A computer database containing the printed word vocabulary of children taking part in the experiment was compiled and used to devise stimuli controlled for grapheme-phoneme correspondence (GPC) frequency and rime neighbourhood consistency according to the children's reading experience. Knowledge of GPC rules and rime units was compared by asking children to read aloud three types of nonword varying in regularity of GPC and consistency of rime pronunciation. Results supported the view that children can acquire knowledge of both GPC rules and rime units from their reading experience. GPC rule strength affects the likelihood of a GPC response; rime consistency affects the likelihood of a rime response. PMID- 10371875 TI - Face naming in dementia: a reply to Hodges and Greene (1998) AB - A case study by Brennen, David, Fluchaire, and Pellat (1996) reported the case of a patient who could occasionally name celebrities with very low concurrent levels of semantic access, which is difficult to reconcile with current models of face identification. Hodges and Greene (1998) attribute Brennen et al.'s case study to artifactual explanations and provide new data, which, they claim, is evidence against the "theory of naming without semantics". This reply demonstrates that Hodges and Greene's arguments are unconvincing and that aspects of Hodges and Greene's data in fact provide support for Brennen et al.'s conclusions. It is also argued that in cases of dementia direct naming has been reported for other stimuli domains as well. PMID- 10371876 TI - Efficacy of a dentifrice containing zinc citrate for the control of plaque and gingivitis: a 6-month clinical study in adults. AB - The objective of this 6-month, double-blind, clinical study, conducted in harmony with American Dental Association (ADA) guidelines, was to evaluate the efficacy of a dentifrice containing 2% zinc citrate and 0.76% sodium monofluorophosphate in a silica base (zinc citrate dentifrice) for the control of supragingival plaque and gingivitis, compared to a control dentifrice containing 0.76% sodium monofluorophosphate in a silica base (control dentifrice). Adult men and women from the Atlanta, Georgia, area were entered in the study and stratified into two treatment groups, which were balanced for baseline Quigley-Hein Plaque Index scores and baseline Loe-Silness Gingival Index scores. Participants received an oral prophylaxis and were instructed to brush their teeth twice daily (morning and evening) for 1 minute with their assigned dentifrice, using a soft-bristled toothbrush. Examinations for supragingival plaque and gingivitis were conducted after 3 months and again after 6 months' use of the study dentifrices. Ninety nine participants complied with the protocol and completed the entire 6-month clinical study. At both the 3- and 6-month study examinations, the zinc citrate dentifrice group exhibited statistically significant reductions in both plaque and gingivitis compared to the control dentifrice group, based on whole-mouth data. At the 6-month examination, the magnitude of these reductions met or exceeded 18% for both plaque and gingivitis (25.3% for plaque; 18.8% for gingivitis). The effect of the zinc citrate dentifrice was most pronounced on the more severe manifestations of plaque and gingivitis, indicating a statistically significant (50.2%) reduction in severe plaque and a statistically significant (66.7%) reduction in severe gingivitis over the control dentifrice after 6 months of use. Similar findings were observed for data obtained from proximal, lingual, and posterior sites. Among the sites that indicated a tendency toward high levels of plaque or gingivitis based on the baseline scores, substantially fewer sites tended to continue to present such high levels at follow-up exams in the zinc citrate dentifrice group than in the control dentifrice group. Thus, in accordance with the 1986 guidelines published by the ADA and the 1994 revision published by the Task Force on Design and Analysis in Dental and Oral Research, the results of this study support the conclusion that a dentifrice containing 2% zinc citrate and 0.76% sodium monofluorophosphate in a silica base is clinically efficacious for the control of supragingival plaque and gingivitis. PMID- 10371877 TI - Clinical efficacy of a dentifrice containing zinc citrate: a 12-week calculus clinical study in adults. AB - The objective of this double-blind clinical study, conducted following the Volpe Manhold design for studies of dental calculus, was to investigate the efficacy of a dentifrice containing 2% zinc citrate and 0.76% sodium monofluorophosphate in a silica base (zinc citrate dentifrice) as compared to a control dentifrice containing 0.76% sodium monofluorophosphate in a silica base (control dentifrice). Adult men and women from the Northern New Jersey area were provided a full oral prophylaxis and assigned the use of a control (non-tartar-control) dentifrice for 8 weeks. At the completion of this initial period, participants were assessed for baseline Volpe-Manhold Calculus Index scores, provided another full prophylaxis, and stratified into two treatment groups that were balanced for age, sex, and baseline calculus. Participants were instructed to brush their teeth twice daily (morning and evening) for 1 minute with their assigned dentifrice, using a soft-bristled toothbrush. Examinations for dental calculus were again performed after 12 weeks' use of the study dentifrices. Seventy-five participants complied with the protocol and completed the entire study. At the 12 week examination, the zinc citrate dentifrice group exhibited a statistically significant 31.9% reduction in mean Volpe-Manhold Calculus Index score as compared to the control dentifrice group. Thus, the results of this clinical study support the conclusion that a dentifrice containing 2% zinc citrate and 0.76% sodium monofluorophosphate in a silica base is efficacious for the control of the development of supragingival calculus. PMID- 10371878 TI - Efficacy of a mouthrinse containing 0.05% cetylpyridinium chloride for the control of plaque and gingivitis: a 6-month clinical study in adults. AB - The objective of this 6-month, double-blind, clinical study, conducted following the American Dental Association (ADA) guidelines, was to provide an assessment of the effectiveness of a newly developed mouthrinse containing 0.05% cetylpyridinium chloride (CPC) for the control of supragingival dental plaque and gingivitis. Adult men and women from the Manchester, England, area were entered in the study, and stratified into two treatment groups (CPC mouthrinse and control mouthrinse), which were balanced for baseline Quigley-Hein Plaque Index scores and baseline Loe-Silness Gingival Index scores. Participants were given an oral prophylaxis and instructed to brush their teeth twice daily (morning and evening) for 1 minute with a soft-bristled toothbrush and fluoride dentifrice provided, immediately followed by rinsing for 30 seconds with 15 cc of their assigned mouthrinse. Examinations for supragingival plaque and gingivitis were conducted after 3 months' and again after 6 months' participation in the study. One hundred eleven participants complied with the protocol and completed the entire 6-month clinical study. At both the 3- and 6-month study examinations, the CPC mouthrinse group exhibited statistically significantly less supragingival plaque and gingivitis than did the control mouthrinse group. At the 6-month examination, the magnitude of these differences met or exceeded 24% for all 4 parameters measured (28.2% for Quigley-Hein Plaque Index, 63.4% for Plaque Severity Index, 24.0% for Loe-Silness Gingival Index, and 66.9% for Gingivitis Severity Index). The magnitude of the reductions in supragingival plaque and gingivitis were adequately large to support a claim of efficacy, in accordance with the criteria provided by the published guidelines of the ADA for the demonstration of the efficacy of a chemotherapeutic agent for the control of supragingival plaque and gingivitis. Thus, the results of this 6-month clinical study support the conclusion that a newly developed mouthrinse containing 0.05% cetylpyridinium chloride provides a statistically significant, clinically relevant level of efficacy for the control of supragingival plaque, and for the control of gingivitis, in accordance with the criteria provided by current ADA guidelines. PMID- 10371879 TI - Local delivery of antimicrobial agents for the treatment of periodontitis. AB - Periodontitis is a result of an infection with specific pathogenic microorganisms. Thus, the local delivery of antimicrobials has been investigated as a possible method for controlling this infection and treating periodontal disease. A number of antimicrobial agents have been studied both as adjunctive therapies with scaling and root planing and as stand-alone chemotherapies. These agents have been administered in irrigation solutions and as single-dose formulations, but with little long-term efficacy in the treatment of periodontitis. Recent investigations have focused on the delivery of antimicrobials in sustained-release formulations designed to maintain effective concentrations of drug within the periodontal pocket. This article provides an overview of the development of the use of locally delivered antimicrobials in periodontal therapy and the current state-of-the-art of the technique. PMID- 10371880 TI - Clinical evaluation of a new 10% carbamide peroxide tooth-whitening agent. AB - NUPRO Gold Tooth Whitening System was evaluated for efficacy according to the proposed ADA guidelines for acceptance. Sixty participants with discolored anterior teeth participated in a 14-day, double-blind, clinical trial. The participants were matched for age, gender, and oral health status and were given either a placebo gel without the active agent or the NUPRO Gold active gel, which they wore in a custom-fabricated mouth guard for home use. The shade of each participant's maxillary anterior teeth was evaluated using a value-oriented Vita Lumin Vacuum Shade Guide before the study. The same shade guide was used to determine shade changes. Time of use of the agent and potential side effects, such as tooth and gingival hypersensitivity and tissue irritation, were assessed at all recall examinations and were recorded by participants in daily diaries. The average shade change for the placebo users was less than one shade. The average shade change for the NUPRO Gold users was 6.96 shades. Tooth hypersensitivity varied from none to severe. Tissue irritation was minimal. The results of these evaluations indicate that NUPRO Gold is effective as a tooth whitening system, when administered properly under the supervision of a dentist, with commonly reported side effects of transient tooth sensitivity and minimal gingival sensitivity. Little or no change in tissue health was noted. This study was supported by Dentsply Preventive Care (York, Pennsylvania). PMID- 10371881 TI - Decay-inhibiting restorative materials: past and present. AB - Clear differences exist in the fluoride release characteristics and setting reactions of glass-ionomer cements and compomers. Differences in decay inhibition associated with specific materials are less clear. Furthermore, resin added to glass ionomer cement formulations and acids added to composite resins make it difficult to distinguish composite resins from compomers and glass ionomer cements, all of which have reported fluoride release. Optimal fluoride release from a dental restorative depends on several conditions, including oral flora, saliva, diet, mineral content of the dental tissues, and marginal seal of the restoration. Presently, in vitro and in vivo studies suggest that materials which behave similarly to silicate cements in their setting reactions and hydration characteristics will behave as decay-inhibiting restoratives. Until optimal fluoride release from dental restoratives can be quantified, dental clinicians are encouraged to consider clinical outcomes as the best test for decay inhibition. Nearly a century of clinical findings support the anticariogenicity of silicate cements. This article reviews fluoride release and anticariogenicity of restorative materials using silicate cement as a model with a well-defined mechanism for preventing secondary caries. The behavior of newer materials is compared to silicate cement for predicting decay inhibition. PMID- 10371882 TI - Forced eruption: a multidisciplinary approach for form, function, and biologic predictability. AB - There are several treatment options for patients with coronal fractures, subgingival caries perforations, and root resorption. Frequently, forced eruption is not considered, although in many cases of single-rooted teeth, forced eruption is the "gold standard" for producing an esthetic result without jeopardizing periodontal support for adjacent teeth. Sufficient tooth length, achieved through forced eruption, ensures the periodontal health of the "biologic width" and crown margin and thus a successful restorative outcome. PMID- 10371883 TI - Prefabricated post-and-core systems: an overview. AB - Restorative dentists should have a working knowledge of more than one post-and core system. This review is designed to encourage them to consider systems they might not have considered otherwise. Many restorative dentists learn to use one prefabricated post-and core system, and they have only one system on hand. When the system on hand is the solution, there is no need for an alternative. However, no single prefabricated post-and-core system can fit all endodontic restorative requirements. The best way to pick an optimum system design is to compare alternatives. The vast assortment of available system components, many supported solely by experimental data without direct clinical corroboration, makes comparison and selection a challenging process. PMID- 10371884 TI - Cryptosporidium: opportunistic environmental water pathogen. PMID- 10371885 TI - Dental dam patch: an effective intraoral repair technique using cyanoacrylate. AB - Secondary dental dam retention is a critical component of successful dental dam isolation and relates to the provision of an effective seal at the dam/tooth junction. Restorative success can be compromised if this seal is inadvertently interrupted during the operative effort. One such periodic mishap is entanglement of the bur and the interdental dam strip during caries or restorative removal. This invariably results in a gaping interproximal defect in the dam. This article discusses the importance of optimum isolation as it relates to current "wet bonding" adhesive procedures, and introduces a repair technique using a patch of dental dam and cyanoacrylate. PMID- 10371886 TI - Long-term prognosis of intentional endodontics and internal bleaching of tetracycline-stained teeth. AB - A total of 112 severely tetracycline-stained teeth in 20 patients were treated by endodontics and nonthermal internal bleaching. All teeth were healthy, intact, and had no history of trauma. The patients were monitored for 5 to 15 years. Excellent, permanent esthetic results were obtained with no side effects. The quality of endodontic treatment and lingual access restoration were important factors in the longevity of the bleaching results. PMID- 10371888 TI - Management of maxillofacial trauma. PMID- 10371887 TI - The use of posts for endodontically treated teeth. PMID- 10371889 TI - Internal fixation of maxillofacial fractures: indications and current implant technologies and materials. AB - Rigid fixation techniques are frequently utilized for the repair of maxillofacial skeletal injuries. This paper reviews the commonly available implants and materials along with their typical indications. The focus is primarily on the most recent trends and developments. Latticework plates and meshes provide greater fixation strength with smaller amounts of metal; modern reconstruction plates allow for fixation of the screws directly to the plates; self-drilling screws make application faster and easier. Resorbable implants made of various polyester polymers are now available, though their indications remain limited thus far. Future developments are discussed as well, including the advent of endoscopic plate placement and image-guided skeletal repositioning. PMID- 10371890 TI - Open reduction and internal fixation of acute mandibular fractures in adults. AB - The mandible is often fractured during maxillofacial trauma because of its prominence within the facial skeleton and the fact that it is a favorite target in interpersonal violence. Treatment of mandibular fractures has made significant advances over the years due to the study of biomechanical principles, advancements in biomaterials and instrumentation and scientifically based research of treatment outcomes. This article will review the principles and philosophy of AO/ASIF techniques as applied to treatment of mandible fractures. Representative cases of different mandible fractures and their treatment will reinforce the clinical application of rigid fixation techniques. PMID- 10371891 TI - Rigid fixation in mandibular reconstruction. AB - A primary goal of mandibular reconstruction is to preserve form and function. Free vascularized bone remains the gold standard for mandibular reconstruction of the anterior defect. While lateral mandibular defects need for vascularized bone remain controversial, free bone flaps provide the most reliable bone stock readily available for dental implantation. Mandibular alignment remains critical for recreating the 3-dimensional relationships of mandibular projection, occlusion, and condylar placement. Depending on the defect, a variety of techniques can be used to maintain mandibular alignment. These include prefabrication of the plate, placement of an internal fixation device, and intermaxillary fixation. Fixation techniques in this day and age generally rely on plates and screws. Miniplates are popular among some reconstructive surgeons while others prefer larger reconstruction plates that can span the entire defect. Bony contouring is critical since keeping bone surfaces in contact allows for improved bone healing. Condylar reconstruction this critical, when the ramus and condylar head are resected, require reconstruction to maintain the entire solid arch. A variety of these techniques are discussed. While the challenge remains reconstituting a solid arch, a variety of techniques can be employed. These techniques including fixation technique, bony contour, mandibular alignment, and condylar reconstruction will be discussed. PMID- 10371892 TI - Pediatric mandibular fractures. AB - Over the last 20 years, a revolution in the management of facial fractures has taken place. Refinements in biocompatible materials of great delicacy and strength along with advances in our understanding of biomechanics of the face, have rendered complex injuries consistently amenable to accurate 3-dimensional reconstruction. Furthermore, with the availability of education in the techniques of internal rigid fixation, these advanced techniques have become routine practice in adults. However, the suitability of rigid internal fixation for children remains controversial. There are many concerns about the effect of implanted hardware in the mandible of a growing child. In addition, some evidence suggests that the elevation of functional matrix off of bone may result in alterations in development. The goal is to restore the underlying bony architecture to its pre-injury position in a stable fashion, with a minimal of aesthetic and functional impairment. However, in children the treatment of bony injuries is most easily accomplished by techniques that may adversely effect craniofacial development. While it is not entirely possible to resolve this dilemma, there exists an extensive body of experimental and clinical information on the appropriate management of pediatric mandibular fractures which can be used to formulate a rational treatment plan for most cases. This paper presents an overview of the contemporary understanding and application of these treatment principles. PMID- 10371893 TI - Internal fixation in pediatric maxillofacial fractures. AB - Pediatric facial trauma presents unique problems in diagnosis and management. The following review highlights relevant points of craniofacial growth and applied anatomy. The epidemiology of pediatric facial fractures is presented followed by pertinent features of clinical presentation and diagnosis. Management of midfacial injuries is discussed individually for each specific type of fracture with emphasis on the role of rigid fixation. Finally, the relationships between childhood fractures, rigid fixation and craniofacial growth are reviewed. PMID- 10371894 TI - Frontal sinus fractures. AB - Frontal sinus fractures can have serious consequences due to the proximity of the sinus to the intracranial cavity, and the potential for nasofrontal duct obstruction with its long-term sequelae. Management of these fractures varies dependent on several factors. These include the degree of displacement of the fractures; involvement of the anterior table, the posterior table, and/or the nasofrontal duct; and the presence of cerebrospinal fluid leakage. This paper gives an overview of fracture treatment options, and offers a simple algorithm for management based on the fracture severity and involvement of the different aspects of the sinus. PMID- 10371895 TI - Naso-orbital-ethmoid complex fracture management. AB - Naso-orbital-ethmoid complex fracture management requires a thorough understanding of the anatomy of medial orbital structures, particularly the medial canthal tendon apparatus. Modern C/T imaging allows one to accurately define the fracture patterns present and to plan the surgical repair. The key structure in these fractures is the "central segment" of bone to which the medial canthal tendon attaches. It is essential to identify, anatomically reduce and fixate this fragment to prevent post operative telecanthus. Standard craniofacial surgical approaches allow the surgeon to widely expose these fractures with minimal secondary cosmetic deformities. Rigid fixation of the fracture segments with titanium microplating systems allows for accurate and stable fixation in a one stage surgical repair. The assessment, imaging, surgical exposure, fixation and care of these fractures will be discussed. The goal of repair is to reestablish a normal intercanthal distance and to anatomically repair the associated fractures. PMID- 10371896 TI - Management of the orbital rim and floor in zygoma and midface fractures: criteria for selective exploration. AB - The management of zygomaticomaxillary and midface fractures has been revolutionized in the past decade as a result of improved surgical access, rigid plating systems, and high-resolution computed tomography. Previously, virtually all midface fractures underwent mandatory orbital exploration to aid in reduction and stabilization. This article emphasizes the importance of reducing and fixating the facial buttresses involved in zygomatic complex fractures, and recommends orbital exploration on a selective basis. Criteria are given to decide which patients require orbital rim and floor exploration. If a patient's fracture does not meet these criteria, the fracture is managed by exposing, reducing, and stabilizing the major facial buttresses involved without performing an orbital exploration. Ninety-seven patients with zygomatic complex fractures were examined and treated with selective orbital rim and floor explorations. Most patients could be managed without the need for orbital exploration, and all were felt to have good fracture reduction and stability. We feel the selection criteria reliably identify patients who do require an orbital exploration and allow the treating surgeon to direct surgical treatment to the site of injury. PMID- 10371897 TI - Management of complex orbital fractures. AB - Fractures of the orbit place the globe and associated structures at significant risk. Management of these fractures requires a thorough ophthalmic evaluation and precise imaging. Contemporary surgical approaches allow the surgeon to repair these injuries with minimal secondary cosmetic deformities and should avoid injury to the globe. A principle goal of therapy is to anatomically reduce fracture segments and to restore a normal orbital volume. Reconstitution of a normal orbital volume and support for the globe will prevent post-injury enophthalmos and globe malposition. The evaluation, imaging, exposure, and repair of orbital fractures is reviewed. Prevention and management of complications is also discussed. The causes of and prevention of blindness is discussed. Application of these techniques should allow for the repair of orbital fractures in a single stage with minimal morbidity. PMID- 10371898 TI - Zygoma complex fractures. AB - The zygomaticomaxillary complex (ZMC) has integral structure and function in the bony skeleton of the face. The prominent convex shape of the ZMC makes it particularly vulnerable to injury. Facial trauma can result in fractures limited to a single buttress of the ZMC but more commonly results in a tetrapod fracture involving all four buttresses. Accurate clinical diagnosis is based on a thorough head and neck examination with ophthalmologic consultation when indicated. Computed tomography is the gold standard for confirmation of the clinical diagnosis and for surgical planning. Multiple surgical approaches to the ZMC are discussed, including the hemicoronal approach to the zygomatic arch, the transconjunctival and subciliary approaches to the orbital rim, and the sublabial approach to the zygomaticomaxillary suture line. The topics of one-, two-, or three-point fixation, as well as plate versus wire fixation, are discussed. Finally, the diagnosis and management of complications such as diplopia and ectropion are discussed. PMID- 10371899 TI - Cranial bone grafting in maxillofacial trauma and reconstruction. AB - Although manipulation of cranial bone has a long history, the use of cranial bone for reconstruction of craniomaxillofacial defects has only become popular in the last 15 years. Many refinements in harvesting and placement techniques have occurred over that time period. Cranial bone is valued for craniofacial skeletal reconstruction because of the proximity of the donor site to the recipient site, reasonable resistance to resorbtion, suitable geometry, and ability to be rigidly fixated. Although intracranial injury is a possibility, careful harvesting techniques minimize donor site morbidity. A detailed knowledge of the anatomy of the tissues around the skull is critical to safe harvesting of cranial bone grafts. Selection of appropriate graft types, proper handling techniques, and use of rigid fixation when indicated result in reliable reconstruction and augmentation of the craniofacial skeleton. PMID- 10371900 TI - Canal configuration of the mesiobuccal root of the maxillary first molar of a Japanese sub-population. AB - AIM: The aim of this study was to determine the percentage of anatomical canal configurations of the mesiobuccal root of the maxillary first molar in Japanese patients. METHODOLOGY: Three hundred teeth were obtained from general dentists who knew absolutely that they were extracted from Japanese patients. The distobuccal and palatal roots were amputated for radiographic convenience. Preoperative radiographs were taken of the remaining crown and mesiobuccal root (MBR) from mesiodistal and buccopalatal directions for each tooth. Routine endodontic access cavities were prepared and size 08 files were placed through the orifice into the MBR until they were seen at the apex. In some cases preparation of the canal orifice with a long shank round burr was necessary to gain access. In seven teeth, no access to the apex was possible and these teeth were eliminated. In the other 293 teeth, the MBR canal(s) was (were) enlarged up to size 15 file. If another canal opening was found at the apex a 08 file was inserted into the second opening and passed coronally. Postoperative radiographs with file(s) in place were taken from the two directions, as before. RESULTS: Of the 293 teeth, 123 (42.0%) were Type II, 89 (30.4%) showed Type III systems and 10 (3.4%) were Type IV. Suggestions for identification and treatment of the second canal in the MBR are presented. CONCLUSIONS: The proportion of cases with two canals in the mesiobuccal root of maxillary first molars from Japanese patients was high and similar to that described from studies of other ethnic populations. PMID- 10371901 TI - 3D computer-aided reconstruction of six teeth with morphological abnormalities. AB - AIM: The purpose of this study was the 3D reconstruction of six teeth with morphological peculiarities using serial cross-sections. METHODOLOGY: All the teeth were put in 3% NaOCl solution after extraction, washed under running water and air-dried. They were then embedded in a two-phase polyester resin and serial cross-sections were produced from each specimen using a special microtome. The thickness of each section was 0.75 mm. Each section was photographed under a stereoscopic microscope. The photographs of the cross-sections were digitized and the external contours of the teeth and the root-canal outline were annotated for each section. Semiautomatic alignment of the sections was achieved with the use of image-processing techniques. Three dimensional surface representation was used in this project to reconstruct the inner and outer surface of the teeth. RESULTS: The results showed in detail the internal morphology of the teeth under investigation. The fact that it was possible to observe and study these teeth from different angles is one of the main advantages of this method as the three dimensional anatomy of these teeth was apparent. CONCLUSIONS: The 3D reconstructing method is a useful tool for the study of the morphology of the teeth. PMID- 10371902 TI - Susceptibility of oral Candida species to calcium hydroxide in vitro. AB - AIM: The susceptibility of common oral Candida species to saturated aqueous calcium hydroxide solution was studied. METHODOLOGY: The yeast species tested were C. albicans (16 strains). C. glabrata (three strains), C. guilliermondii (three strains), C. krusei (two strains), and C. tropicalis (two strains). At least one reference strain of each species was used; the others were clinical isolates either from persistent apical periodontitis or from marginal periodontitis. The susceptibility of Enterococcus faecalis (ATCC 29212) was studied for comparative purposes. Standardized inocula of the strains were incubated in aqueous calcium hydroxide solution, pH 12.4, for time-periods ranging from 5 min to 6 h. Volumes of 0.1 mL of the test suspension were cultured directly on Brucella blood agar and incubated in air at 37C. The plates were inspected for growth at 24 and 48 h and the colonies were counted. The time required to reduce the number of colony-forming units to less than 0.1% of the initial number was determined for each strain. RESULTS: The sensitivity of the C. albicans strains was generally low, with 16 h of incubation required to kill 99.9% of the colony-forming units. No differences in susceptibility between C. albicans strains isolated from root-canal infections and from periodontitis were found. Both strains of C. tropicalis were killed between 3 and 6 h of incubation, whilst strains of C. guilliermondii were killed after only 1020 min of incubation. All strains of C. glabrata survived 20 min, but not 1 h, of incubation, whilst 13 h were required to kill C. krusei. Compared with E. faecalis, all Candida spp. showed either equally high or higher resistance to aqueous calcium hydroxide. CONCLUSIONS: This study indicates that Candida spp. are resistant to calcium hydroxide in vitro, which may explain the isolation of yeasts from cases of persistent apical periodontitis. PMID- 10371903 TI - Antimicrobial effects of various endodontic irrigants on selected microorganisms. AB - AIM: This study was undertaken to determine the antimicrobial effect of various endodontic irrigants against six selected microorganisms. METHODOLOGY: Staphylococcus aureus, Enterococcus faecalis, Streptococcus salivarius, Str. pyogenes, Escherichia coli and Candida albicans were included in the study. Pre sterilized Whatman paper discs, 6 mm in diameter and soaked with the test solution, were prepared and placed onto the previously seeded agar Petri plates. Each plate was incubated aerobically. A zone of inhibition was recorded for each plate and the results were analysed statistically. RESULTS: 5.25% NaOCl was effective against all test microorganisms with a substantial zone of inhibition. Saline was always ineffective. Decreased concentration of NaOCl significantly reduced its antimicrobial effect. Cresophene showed a significantly larger (P < 0.05) average zone of inhibition compared to the other experimental irrigants. Alcohol had smaller but not significantly different zones of inhibition than chlorhexidine. CONCLUSIONS: 5.25% NaOCl was superior in its antimicrobial abilities compared with other irrigants used. A reduced concentration of NaOCl (0.5%) resulted in significantly decreased antimicrobial effects. When compared with 21% alcohol, 0.5% NaOCl and 2% chlorhexidine, paramonochlorophenol (cresophene) showed a greater antimicrobial effect. PMID- 10371905 TI - Location of contact areas on rotary Profile instruments in relationship to the forces developed during mechanical preparation on extracted teeth. AB - AIM: The aim of this study was to locate the areas of direct instrument contact with dentine in the root canal system during rotary preparation and to analyse the relationship between these areas and the vertical forces and torque developed during the preparation. METHODOLOGY: Canal preparations were performed by endodontists either with the step-back (SB) or the crown-down (CD) technique. In order to locate the areas of contact, the instruments were coated with two layers of gold by electro-deposition. They were photographed before and after use, and a coding system, based on mm from the instrument tip, was devised to designate areas of gold removal or instrument wear due to friction. To standarise the conditions of instrument manipulation, the teeth were fixed in the Endographe holder, and this device was used to measure vertical forces and torque. RESULTS: The results showed that the first series of instruments used for the CD technique (taper 0.06) left 2 +/- 1 mm of the tip with the gold intact, indicating that these instruments and this step of the CD technique are the safest part of the preparation. For all other instruments (taper 0.04 series), the areas of gold removal involved the 3 mm around the tip and this finding was independent of the order of instrument use and preparation technique (SB or CD). The differences between the two techniques were significant in terms of the mean area of decolouration and the mean force and torque values. For the SB and CD techniques, the contact areas were, respectively, 10 +/- 3 and 7 +/- 2 mm. The forces and torque values were correspondingly higher for SB vs. CD; the mean values were, respectively, 19 and 21 N for vertical forces and 16 10(5) and 13 10(5) Nm for torque. CONCLUSIONS: The recorded torque values and the location on the instruments of the areas of contact with dentine during this development of torque i.e. at or near the tip, indicate that great caution should be used with the rotary technique, particularly with the taper 0.04 instruments, regardless of preparation technique. PMID- 10371904 TI - Comparison of measurements obtained with hand files or the Canal Leader attached to electronic apex locators: an in vitro study. AB - AIM: The aim of the present in-vitro study was to combine directly the Canal Leader handpiece (SET, Olching, Germany) with the electronic apex locators ROOT ZX (Morita, Kyoto, Japan) and JUSTY (Yoshida, Tokyo, Japan) to find out whether the working length values thus obtained were identical to those resulting from the combination of the same electronic devices with hand files. METHODOLOGY: A total of 50 natural extracted teeth with single canals and mature apices were used. A radiograph was used as a control and the distance from the radiographic apex to the tip of the file was measured and compared with the results of the electronic length determination. RESULTS: For both electronic devices the differences amongst the distribution of the measurements were not statistically significant under the specified conditions (P > 0.05), indicating that the measurements with hand files and with the Canal Leader were identical for the majority of the cases. CONCLUSIONS: Under the conditions of this study the working length of canals obtained with electronically assisted hand files were similar to those obtained with the electronically assisted mechanical handpiece Canal Leader. PMID- 10371906 TI - Cyclic fatigue of Profile rotary instruments after simulated clinical use. AB - AIM: The purpose of this study was to evaluate cyclic fatigue of Profile Ni-Ti rotary instruments (PRIs) after dry heat sterilization and up to 10 times simulated clinical use. METHODOLOGY: Instruments of size 40-15 were used in a crown-down technique. Three groups were included in this study. In groups 1 and 2, each set of instruments was used in five and 10 canals, respectively. Group 3 was the control group. NaOCl at a concentration of 2.5% was used as an irrigant. Each set of instruments was sterilized before each use. RESULTS: The PRI size 40 demonstrated the lowest incidence of rotations to breakage. One-way analysis of variance and Turkey's HSD test showed statistically significant differences among different file sizes within each group. CONCLUSIONS: The results showed that dry heat sterilization and simulated clinical use in the presence of NaOCl did not lead to a decrease in the number of rotations to breakage of the files. PMID- 10371907 TI - In vitro evaluation of the reliability of the Root ZX electronic apex locator. AB - AIM: An experimental comparison of the Root ZX apex locator with the real measurement of the root canal length was carried out using a saline gel to simulate the periodontium. METHODOLOGY: Lengths were taken when the needle reached the 0.5 mark and the Apex mark on the meter. These measurements were then compared with a calculated reference length representing the real length of the root canal. Reliability was assessed using Student's t-test. Precision was assessed using the mean and standard deviation of differences. RESULTS: The Root ZX gave measurements within a range of 0.5 mm in 84.72% of the cases. The intra operator and inter-operator variabilities were not statistically significant (P > 0.05). CONCLUSIONS: The Root ZX is not capable of detecting the '0.5 mm from the foramen' position and thus, should only be used to detect the foramen (major diameter). PMID- 10371908 TI - Inducible nitric oxide synthase activity by interferon-gamma-producing cells in human radicular cysts. AB - AIM: In this study, the interaction of interferon-gamma-(IFN-gamma) and inducible nitric oxide synthase (iNOS)-producing cells in human radicular cysts were investigated. METHODOLOGY: Inflamed periapical tissues were obtained from patients at the time of endodontic surgical treatments and were cut into two pieces. After fixing with acetone or 4% paraformaldehyde in phosphate-buffered saline, 5-m-thick paraffin and cryostat sections were prepared. The paraffin sections of the inflamed tissues were evaluated histologically with haematoxylineosin stains. The specimens diagnosed as radicular cysts were then examined by immunostaining. Immunohistochemistry for iNOS and fluoresence microscopy for IFN-gamma using the cryostat sections were performed with a mixture of affinity purified human iNOS antiserum and human IFN-gamma monoclonal antibodies. RESULTS: The results revealed that iNOS-gamma producing cells localized adjacent to IFN-gamma-producing cells. In addition, some of iNOS producing cells exhibited immunoreactive IFN-gamma. On the other hand, epithelial cells showed significant levels of iNOS production, but not IFN-gamma. CONCLUSIONS: The data would suggest the possibility that iNOS production could be precisely controlled by autocrine or paracrine effects of IFN-gamma producing cells in radicular cysts and might play a pivotal role in periapical lesions. These findings are consistent with a hypothesis suggesting that NO inhibitors could be used through the root canals as a pharmacological treatment for periapical lesions. PMID- 10371909 TI - Apical transportation revisited or 'where did the K-file go'? AB - CASE REPORT: This case report describes the outcome of a number of retreatments on a failed root filling in a maxillary first molar. The patient wanted all amalgams replaced by tooth-coloured Cerec restorations, including one in a symptomless maxillary molar. This tooth had a pulpotomy or a poorly done root canal treatment 10 years earlier. The molar was root-canal retreated before placing the Cerec restoration and the palatal canal was filled 5 mm short of the radiographic apex. About 1 year later the patient presented with pain. Suspecting that a second mesiobuccal canal (MB-2) had not been located, a second non surgical retreatment was instituted. MB-2 was not found and the palatal canal was retreated a third time, setting the working length 2 mm short of the radiographic apex. Because pain persisted palatally an apicectomy was performed and the tooth became symptomless. The resected palatal root apex was subsequently serially cross-sectioned, photographed and the canals analysed. Obvious apical transportation occurred during the cleaning and shaping procedures. Analyses of the canals showed that despite the retreatments, 11% of the canal cross-sectional area remained uncleaned although 7% of the root area was 'shaped'. Radiographically, the obturated palatal canal appeared reasonably well centred. However, this was disproved by the cross-sections, indicating that in this case, the clinician did not know where the K-Files had 'gone'. Apically, the obturated canal was certainly not within the natural canal. The pain located palatally was probably due to inadequate cleaning and shaping of the apical part of the root canal and its accessory canals. PMID- 10371910 TI - Thermal sensitivity of endodontically treated teeth. AB - CASE REPORTS: The problem of thermal sensitivity following non-surgical root canal treatment is explored and case reports are presented. Possible causes for post-treatment discomfort from endodontic and restorative aetiologies are discussed, as are the mechanisms to explain the patients' painful experiences. Treatment of this problem may vary from the simple replacement of a defective restoration to a more extensive non-surgical retreatment of the case, despite radiographic evidence of an acceptable root filling and normal periradicular tissues. PMID- 10371911 TI - Surgical extrusion of crown-root-fractured teeth: a clinical review. AB - AIM: In this clinical study combined surgical and endodontic treatment was performed in 20 cases of crown-root fracture and the outcomes reviewed. METHODOLOGY: Surgical treatment involved a conventional extraction and stabilization technique. Root canal treatment using calcium hydroxide was performed. Before root canal obturation, a calcium hydroxide dressing was maintained for 3 months. RESULTS: Follow-up examinations, which varied between 6 and 36 (mean 14.5) months, showed that there were no radiographic and clinical signs of progressive root resorption, marginal bone loss or periapical disease in all except one case. CONCLUSIONS: The favourable results of this study demonstrate that surgical extrusion in teeth with crown-root fractures may be an alternative treatment to orthodontic extrusion. PMID- 10371912 TI - Porcelain veneers: dentin bonding optimization and biomimetic recovery of the crown. AB - PURPOSE: The purpose of this study was to investigate the biomimetic principle in porcelain veneer reconstruction, or in other words, to assess the extent to which the restoration can mimic the biomechanics and structural integrity of the original tooth. Using an optimized luting procedure, porcelain veneers are expected to present such features even when bonded to an extensive dentin surface. METHODS AND MATERIALS: Dentin-bonded porcelain veneers were assessed using functional and cyclic thermal loads with respect to two parameters: coronal stiffness (investigated using experimental strain gauges and finite element analysis) and morphology of the tooth-restoration interface (scanning electron microscope evaluation). Two different application modes of the same dentin bonding agent, Optibond FL, were evaluated: a traditional method (dentin adhesive applied when proceeding to luting the veneer) and an alternative method (dentin adhesive applied to dentin and cured before taking the impression for the veneer). RESULTS: In the finite element model, the crown compliance increased by a factor of 2.16 after facial enamel removal and returned to 96% of its original value after the placement of the veneer. The finite element values showed a good correlation with strain gauge experimental results (one-sample t test, P > 0.35 after facial enamel removal and P > 0.19 after veneer placement). The dentin adhesive application mode was not critical to the recovery of tooth stiffness (analysis of variance, P = 0.10). However, qualitative scanning electron microscope observations demonstrated that the traditional dentin adhesive application was associated with bonding failures between the hybrid layer and the overlying resin, whereas unbroken and continuous interfaces were obtained with the new method using the same dentin adhesive. CONCLUSION: The results of this study definitely favor the biomimetic behavior of porcelain veneers bonded to teeth using an optimized application mode of dentin adhesives, because this treatment modality proved to restore both the mechanical behavior and microstructure of the intact tooth. PMID- 10371913 TI - Clinical examination of leucite-reinforced glass-ceramic crowns (Empress) in general practice: a retrospective study. AB - PURPOSE: The purpose of this study was to retrospectively evaluate leucite reinforced-glass ceramic crowns (Empress) placed in patients who regularly visit general practices. MATERIALS AND METHODS: One hundred ten Empress crowns, placed in 29 patients who visited a general practice on a regular basis, were evaluated according to the California Dental Association's (CDA) quality evaluation system. In addition, the occurrence of plaque and certain gingival conditions was evaluated. All crowns were luted with resin composite cement. The mean and median years in function for the crowns were 3.6 and 3.9 years, respectively. RESULTS: Based on the CDA criteria, 92% of the 110 crowns were rated "satisfactory." Eighty-six percent were given the CDA rating "excellent" for margin integrity. Fracture was registered in 6% of the 110 crowns. Of the remaining 103 crowns, the CDA rating excellent was given to 74% for anatomic form, 86% for color, and 90% for surface. No significant differences (P > 0.05) were observed regarding fracture rates between anterior and posterior crowns. With regard to the occurrence of plaque and bleeding on probing, no significant differences (P > 0.05) were observed between the Empress crowns and the controls. CONCLUSION: Most of the fractured crowns had been placed on molars or premolars. Although the difference between anterior and posterior teeth was not statistically significant with respect to the fracture rates obtained, the number of fractured crowns placed on posterior teeth exceeded that of those placed on anterior teeth. The difference between the fracture rates may have clinical significance, and the risk of fracture has to be taken into consideration when placing crowns on teeth that are likely to be subjected to high stress levels. PMID- 10371914 TI - Unpolymerized layer on autopolymerizing, hard reline materials. AB - PURPOSE: The purpose of this study was to clarify certain properties of the resulting unpolymerized surface layer on 6 hard, autopolymerizing reline resins. MATERIALS AND METHODS: Two of the resins examined were the conventional type with a base monomer composition of mainly methyl methacrylate, and the other four were cross-linked reline materials containing difunctional monomers. For curing, the materials were polymerized in air at 24 and 37 degrees C, and in distilled water at 37 degrees C. The powder-to-liquid ratio was changed by +/- 20 wt% of the manufacturer's specified ratio. The inhibition depth processed under these conditions was measured with an optical microscope. Each sample was then immersed in methylene blue dye bath for 3 weeks before its surface was observed. RESULTS: Especially on cross-linked reline materials, the unpolymerized layer was significantly reduced with higher temperature, lower oxygen presence, and lower powder-to-liquid ratio (P < 0.01). The unpolymerized layer was stained in all materials. There were tiny, stained voids in the polymerized region on the conventional type of reline resins, whereas no staining was found in the polymerized region on the cross-linking reline resins. CONCLUSION: The inhibition depth was strongly affected by the temperature, the presence of air, and viscosity. The unpolymerized layer was easily contaminated in all materials. However, the highly polymerized, cross-linked reline materials might be harder to contaminate than the conventional type of reline resins. PMID- 10371915 TI - A longitudinal clinical study of Procera ceramic-veneered titanium copings. AB - PURPOSE: The purpose of the present paper was to study the long-term clinical results with ceramic-veneered Procera titanium copings. MATERIALS AND METHODS: A total of 44 titanium copings (fabricated for 22 patients) veneered with a low fusing ceramic were followed for 60 to 78 months. The clinical examinations were performed by licensed specialists in prosthetic dentistry. The crowns were rated according to the California Dental Association system. In addition, Bleeding Index and Margin Index were also evaluated. RESULTS: In 3 crowns ceramic fractures necessitated their replacement. Two crowns had to be replaced because of caries. The ratings for surface and color had changed markedly, from excellent to acceptable. Regarding anatomic form, with the exception of the 3 fractured ceramic crowns, there were no obvious changes. The margin integrity, aside from the 2 crowned teeth with caries, was recorded as satisfactory (excellent or acceptable) for all other crowns; in fact, a large majority were rated excellent. Regarding Bleeding Index, there were no differences between crowned teeth and control teeth. Changes in Margin Index showed that the gingiva of the crowned teeth had retracted. CONCLUSION: Of the various clinical factors evaluated, only surface and color-related to the low-fusing ceramic used for veneering-showed any obvious change during the follow-up period. Otherwise the veneered titanium copings had, in general, performed well. PMID- 10371916 TI - Color stability of visible light-cured, hard direct denture reliners: an in vitro investigation. AB - PURPOSE: Previous studies have disclosed the unsatisfactory color stability of autopolymerizing, hard direct denture reliners (HDDR). The present study investigated the color stability of the newly introduced visible light-cured and dual-cured HDDRs. MATERIALS AND METHODS: Five HDDRs were evaluated after 1 hour, 1 day, 7 days, and 30 days of immersion in coffee, tea, and water. Color measurements were obtained with a tristimulus colorimeter, and color differences (delta E*) were calculated. RESULTS: After 30 days of immersion one visible light cured material exhibited the highest delta E* value (17.8). The other materials showed acceptable color stability in water and coffee. However, tea strongly affected their color. CONCLUSION: Reliners, staining solutions, and immersion time are significant factors that affect color stability. After 7 days of immersion, all of the materials showed perceptible color differences. Tea exhibited higher staining capacity than coffee. PMID- 10371917 TI - Displacement of mandibular removable partial denture bases by tongue movements during speech. AB - PURPOSE: The purpose of this study was to evaluate denture base displacement caused by tongue movements during speech. MATERIALS AND METHODS: Ten patients with new Kennedy Class I mandibular removable partial dentures participated in this study. All prostheses were made according to a standardized protocol. Denture base displacements when speaking a test sentence were measured with a computer-aided device that was developed for this purpose. It included 4 Hall sensors, mounted on a paraocclusal splint and magnets, embedded in the denture bases. Vertical and transverse base displacements were calculated from Hall voltages. Clasp retention was measured with a dynamometer. RESULTS: Mean displacements were 145 microns seating, 72 microns unseating, 87 microns buccally, and 74 microns lingually. Seating and unseating displacements differed statistically significantly in absolute values. Mean clasp retention was 2.5 N. No correlation was found between clasp retention and unseating base displacements. CONCLUSION: Tongue movements during speech induced only minimal displacements in the dentures investigated. Additionally, tongue activity was found to be denture stabilizing rather than denture displacing in this study. PMID- 10371918 TI - A review of in vitro and in vivo methods to evaluate the efficacy of denture cleansers. AB - PURPOSE: This review summarizes the methods employed to evaluate denture cleansers and makes some suggestions on the methodology of evaluation. MATERIALS AND METHODS: More than 20 articles evaluating the efficacy of denture cleansers were compared, and the advantages and disadvantages of each method were evaluated. RESULTS: The results obtained vary depending on the methods used to evaluate the efficacy of denture cleansers, particularly among in vitro and in vivo assays. In addition, it is pointed out that chemical denture cleansers are not as efficacious in clinical use as in the in vitro assay. The uncertainty over efficacy may be caused by nonstandardized methodology and reports of conflicting results. CONCLUSION: A standardization of the methodology is needed. This should include evaluation of the efficacy of cleansers through both in vivo and in vitro assay methods; standardization of the materials and methods used in studying the efficacy of denture-cleansing regimens; and the examination of not only selected microorganisms from limited surface areas of a denture, but the microorganisms in denture plaque from the whole surface of a denture. It would also be preferable to use methods and/or media that make it possible to not only qualify but also quantify the microbial plaque. PMID- 10371919 TI - Impact of implant interdependency when evaluating success rates: a statistical analysis of multicenter results. AB - PURPOSE: When evaluating the outcome of oral implant treatment, a statistically important aspect to consider is whether any dependency exists among implants placed within the same patient/jaw. If one implant fails, will the risk of subsequent failures increase, i.e., will any of the remaining implants also fail? In an attempt to study this question, the aims of the present study were to statistically determine if the suggested hypothesis was valid, i.e., whether dependency exists among implants in the same patient/jaw, and to determine how failure lifetable analysis should be calculated. MATERIALS AND METHODS: In the present study, multicenter trial material consisting of 1.738 oral implants in 487 patients from 4 separate studies was used. First, any dependency among implants within the same patient/jaw was determined; thereafter, cumulative success rates were calculated for one implant per patient (chosen randomly), and the range of variations in the randomized success rates was also calculated. This was then compared with the cumulative success rates based on the entire material. RESULTS: The statistical analysis showed that a dependency among the implants existed prior to their functional loading, i.e., the risk for an implant failure among the remaining implants in the same patient/jaw increased after the first failure had occurred. CONCLUSION: Both study design and statistical analysis are of importance when comparing success rates from various investigations, since dependency among implants in the same patient/jaw does exist and may influence the success rates. Consequently, it is suggested that only one randomly selected implant from each patient should be considered when calculating implant success rates. PMID- 10371920 TI - Strategies to achieve fit in implant prosthodontics: a review of the literature. AB - PURPOSE: This paper reviews the literature on advanced strategies that attempt to improve fit in implant prosthodontics with reference to the concept of the "distortion equation." MATERIALS AND METHODS: The majority of the articles reviewed were either clinical or technique articles that advocated strategies to improve fit in implant prosthodontics. A limited number of retrospective and prospective clinical trial studies were included as they related to the topic. Reviewed articles were limited to those that addressed advanced strategies to improve fit. All of the scientific studies included in this review used an in vitro experimental design. The advanced strategies were categorized into methods that address intraoral indexing and methods that use the implant master cast. RESULTS: Relatively few methods have been scientifically proven to improve fit in implant prosthodontics. Most of the tested strategies still resulted in a slight misfit of the frameworks to the implant abutments/analogues. CONCLUSION: Multiple factors preclude that the concept of "passive fit" can be achieved in implant prosthodontics, even with the use of advanced strategies. The use of meticulous, accurate implant prosthodontic procedures and the appropriate use of advanced strategies continue to be the recommended means of achieving precise fit of the implant prosthesis to the intraoral abutments. PMID- 10371921 TI - Influence of ozone on oxidation of dental alloys. AB - PURPOSE: The purpose of this study was to examine the influence of ozone on the surface of removable partial denture (RPD) alloys to determine its usefulness as a cleaning method for RPDs, since ozone has powerful sterilizing and deodorizing properties. MATERIALS AND METHODS: Two types of ozone cleaning were used. The quantities of ozone generated by both methods were the same (20 mg/h). In method A, ozone was generated for 10 minutes every 12 hours and in method B, ozone was generated over 24 hours a day. Test specimens of 3 types of dental alloy (Co-Cr, Au-Ag-Pt, and Au-Cu-Ag-Pd) were subjected to different cleaning methods for 7 days and measured in terms of reflectance, surface roughness, and weight. Five different cleaning solutions (three commercial denture cleaners, acid electrolyzed water with a pH of 2.4, and pure water) were used for comparison with the ozone treatments. RESULTS: No significant changes were detected after treatment of the Co-Cr and Au-Ag-Pt alloys with ozone. Ozone caused a slight change in the Au-Cu-Ag-Pd alloy in terms of reflectance, but the changes were significantly less than those caused by acid-electrolyzed water and one of the commercial denture cleaners. CONCLUSION: Ozone had little influence on the oxidation of dental alloys. PMID- 10371922 TI - Optimal esthetics in single-tooth replacement with the Re-Implant system: a case report. AB - PURPOSE: This report demonstrates the use of root-analogue titanium implants for single-tooth replacement. MATERIALS AND METHODS: A maxillary lateral incisor was removed and a custom-made one-stage, root-analogue titanium implant (Re-Implant) with an apical extension and a healing cap was fabricated and placed immediately after tooth extraction. Six months later the healing cap was removed, an impression was taken, and a porcelain-fused-to-metal crown was fabricated and cemented. RESULTS: No complications occurred during the healing period. A good esthetic result was achieved with the final ceramometal crown. Bony resorption and buccal soft tissue recession led to a slight discoloration of the marginal periimplant mucosa. CONCLUSION: Further research into the modalities of the immediate placement of root-analogue implants is needed to obtain predictable esthetic results concerning the soft tissue environment. PMID- 10371923 TI - Better service vs. self-service. PMID- 10371924 TI - Managed care or commercial care? PMID- 10371925 TI - The law and managed care plans. PMID- 10371926 TI - Fee-for-service dentistry in an era of managed care. PMID- 10371928 TI - A viable benefits alternative. PMID- 10371927 TI - Beyond capitation--a closer look at dental HMOs. PMID- 10371929 TI - Managed care--a personal perspective. PMID- 10371930 TI - Impact on practice value. PMID- 10371931 TI - Who knows the difference? PMID- 10371932 TI - Managed care and healthcare reform. Interview by James Pride. PMID- 10371933 TI - Subsidized smiles, clean eyes and more. PMID- 10371934 TI - Distraction osteogenesis [introduction]. PMID- 10371935 TI - Distraction osteogenesis of the mandible: a ten-year experience. AB - Mandibular distraction has been performed at the authors' institution for the past 10 years on a variety of craniofacial anomalies. This article reviews the experience with distraction and outlines the authors' treatment algorithms based on patient age and pathology. The roles of distraction versus conventional orthognathic surgery are reviewed. The need for preoperative surgical planning and postoperative orthodontic therapy is emphasized. PMID- 10371936 TI - Treatment planning and biomechanics of distraction osteogenesis from an orthodontic perspective. AB - As in traditional combined surgical and orthodontic procedures, the orthodontist has a role in the planning and orthodontic support of patients undergoing distraction osteogenesis. This role includes predistraction assessment of the craniofacial skeleton and occlusal function in addition to planning both the predistraction and postdistraction orthodontic care. Based on careful clinical evaluation, dental study models, photographic analysis, cephalometric evaluation, and evaluation of three-dimensional computed tomographic scans, the orthodontist, in collaboration with the surgeon, plans distraction device placement and the predicted vectors of distraction. Both surgeon and orthodontist closely monitor the patient during the active distraction phase, using intermaxillary elastic traction, sometimes combined with guide planes, bite plates, and stabilization arches, to mold the newly formed bone (regenerate) while optimizing the developing occlusion. Postdistraction change caused by relapse is minimal. Growth after mandibular distraction is variable and appears to be dependent on the genetic program of the native bone and the surrounding soft tissue matrix. A significant advantage of distraction osteogenesis is the gradual lengthening of the soft tissues and surrounding functional spaces. Distraction osteogenesis can be applied at an earlier age than traditional orthognathic surgery because the technique is relatively simple and bone grafts are not required for augmentation of the hypoplastic craniofacial skeleton. In this new technique, the surgeon and the orthodontist have become collaborators in a process that gradually alters the magnitude and direction of craniofacial growth. PMID- 10371937 TI - Orthodontic management of the patient undergoing mandibular distraction osteogenesis. AB - Distraction osteogenesis is a gradual incremental bone-lengthening technique that requires very precise treatment planning and surgical execution. Subtle variation in the position of the corticotomy/osteotomy or in the position of the distraction device will affect the ultimate position of the tooth-bearing (distal) segment. Other functional and/or anatomic factors (that at this time are not well understood) also have an influence on the direction in which the tooth bearing segment moves during distraction. These factors combine to create a discrepancy between the planned direction and the actual or observed direction of distraction. The charge for the orthodontist is to plan, as carefully as possible and in concert with the surgeon, the direction (vector) in which the tooth bearing segment will travel during distraction. Further, the orthodontist must be able to modify the position that the tooth-bearing segment takes by exerting orthopedic and orthodontic forces and by making adjustments to the distraction device. In addition, the orthodontist must prepare the dentition before distraction, manage the dentition during distraction, and finish the occlusion after distraction in a manner that is different from more traditional jaw respositioning techniques. PMID- 10371938 TI - Intraoral mandibular distraction osteogenesis. AB - During recent years, distraction osteogenesis has gained in popularity for the treatment of various bone deficiencies either in the vertical, transverse, or anteroposterior dimension. Distraction osteogenesis has been shown to be an effective technique for mandibular widening and lengthening where traditional orthognathic surgery has important limitations. The intraoral approach to these procedures prevents damage to the inferior alveolar nerve and the developing dental follicles, and eliminates hypertrophic facial scars. Intraoral distraction osteogenesis also avoids donor-site morbidity, and minimizes the need for blood transfusion or prolonged fixation. This intraoral application provides for enhanced patient acceptance and reduces the potentially negative psychosocial effects of wearing an extraoral distraction appliance. PMID- 10371939 TI - Combined maxillary and mandibular distraction osteogenesis. AB - Mandibular elongation by gradual distraction in patients with hemifacial microsomia is a simple and effective procedure to correct facial asymmetry. The changes in mandibular dimension result in changes in dental occlusion. These are minimal in children because of the rapid growth of the maxilla and can be corrected easily with minor orthodontic treatment. Mandibular distraction in adults with hemifacial microsomia produces good aesthetic results but leaves the patient with a severe alteration in the occlusion requiring complex orthodontic treatment over a long period of time. To avoid this problem, an incomplete Le Fort I osteotomy is performed simultaneously with the mandibular corticotomy. Intermaxillary fixation is placed on the fifth postoperative day, and distraction is initiated. This technique preserves the preexisting stable occlusion. After distraction, both the maxillary and mandibular occlusal planes become horizontal, and facial asymmetry is corrected. PMID- 10371940 TI - Maxillary distraction for the management of cleft maxillary hypoplasia with a rigid external distraction system. AB - Maxillary hypoplasia is a common finding in patients with repaired orofacial clefts. Management of this condition has been a challenge to the reconstructive team. The introduction of distraction osteogenesis to treat craniofacial skeletal dysplasias has opened alternative approaches to manage these severe conditions. In this article, the authors present their technique to distract the hypoplastic cleft maxilla using a rigid external distraction device. The clinical assessment, indications, orthodontic procedure, surgical technique, and distraction protocol are reviewed. A case report shows the use of the technique. This technique allows the reconstructive team to treat patients in all age groups with predictable and stable results. PMID- 10371941 TI - Midface distraction. AB - Since the initial application of distraction osteogenesis to the human mandible by McCarthy, distraction osteogenesis has been used for gradual lengthening of the midface in children with syndromic craniosynostosis, cleft lip and palate, hemifacial microsomia, and midface hypoplasia from other causes. Both external and internal devices are available that permit midface distraction. The background of midface distraction and the development of a Modular Internal Distraction (MID) system that permits widespread use of easily customized, buried distraction devices throughout the craniofacial region are presented. The relative and potential clinical indications for distraction, treatment planning, patient preparation, and possible surgical orthodontic interactions during distraction, as well as a variety of case examples showing the MID system, are discussed. PMID- 10371942 TI - Reconstruction of a neocondyle using transport distraction osteogenesis. AB - The process of slow bone expansion by distraction osteogenesis in conjunction with functional remodeling can also be used for the reconstruction of a neomandible and neocondyle. This is the technique of transport distraction osteogenesis. A transport disc is surgically created adjacent to the area of a discontinuity defect, and the transport disc is advanced by the process of distraction osteogenesis, using the Ilizarov principles. The mandible therefore acts as the bony template for reconstruction such that the neomandible created from the distraction process has the same size and shape as the native mandible covered by gingiva. This allows for enhanced prosthetic reconstruction. A reverse L osteotomy of the ramus can also be performed to create a transport disc to reconstruct a neocondyle. Because the leading edge of the transport disc becomes enveloped by a fibrocartilagenous cap, the ramal transport disc can be moved superiorly to create a new articulation. Patients are encouraged to open and close their mouths during the distraction process, such that the transport disc remodels to form a neocondyle. This technique was successfully used to treat patients with degenerative joint disease, condylar resorption, and bony ankylosis. PMID- 10371943 TI - Review of devices for distraction osteogenesis of the craniofacial complex. AB - Over the last 50 years, Ilizarov refined a method to successfully lengthen endochondral bones and the surrounding soft tissue matrix. Given the difficulties in reconstructing deformities of the craniofacial complex, distraction osteogenesis has recently been used to avoid the problems associated with conventional surgery and to begin correction at an earlier age. Distraction devices can be categorized by whether they are internal or external, the direction of distraction, and the site of application. External devices are capable of either unidirectional, bidirectional, or multiplanar (three dimensional) distraction. Internal or intraoral distractors are capable of unidirectional distraction only. Distraction devices used to lengthen the mandibular ramus and body, widen the mandible, augment the alveolar ridge, conduct bone transport, and advance the midface are reviewed. PMID- 10371944 TI - The many contributions of organ transplantation to science and humanity. PMID- 10371945 TI - Current status and future prospects for organ procurement and retrieval. PMID- 10371946 TI - What should be the role of the state in the development of transplantation? PMID- 10371947 TI - Living nonrelated kidney transplantation: time to be taken seriously. PMID- 10371948 TI - Nonimmunologic matching in renal transplantation. PMID- 10371949 TI - Mechanism and prevention of ischemia-reperfusion injury. PMID- 10371950 TI - Present and future challenges in living related liver transplantation. PMID- 10371951 TI - What should be the role of the state in the development of transplantation? PMID- 10371952 TI - Pediatric renal transplantation. PMID- 10371953 TI - Pathophysiology and diagnosis of acute rejection. PMID- 10371954 TI - Therapeutic strategies for optimal use of novel immunosuppressants. PMID- 10371955 TI - Current knowledge of the pathogenesis of chronic allograft dysfunction. PMID- 10371956 TI - Are there markers to initiate treatment of chronic rejection? PMID- 10371957 TI - Chronic rejection: new potential venues of therapy. PMID- 10371958 TI - Clinical application of tolerance induction in solid organ transplantation. PMID- 10371960 TI - Clinical aspects of immunologic monitoring. PMID- 10371959 TI - Regulation of immune reactivity and tolerance by antigen migration and localization: with particular reference to allo- and xenotransplantation. PMID- 10371961 TI - Potential of gene transfer in transplantation. PMID- 10371962 TI - Barriers to xenotransplantation. PMID- 10371963 TI - The Rapaport-Starzl Festschrift: a summation. PMID- 10371964 TI - HLA-G genetic polymorphism in 57 Portuguese white families studied by PCR-RFLP and PCR-SSOP. PMID- 10371965 TI - HLA-G gene polymorphism in the normal population and in recurrent spontaneous abortion in Hungary. PMID- 10371966 TI - Detection of soluble HLA-G levels in maternal serum can be predictive for a successful pregnancy. PMID- 10371967 TI - Natural killer cell reactivity and HLA-G in recurrent spontaneous abortion. PMID- 10371969 TI - A nonhuman primate model for maternal-fetal immune tolerance. PMID- 10371968 TI - Determination of soluble HLA-G and HLA-A, -B, and -C molecules in pregnancy. PMID- 10371970 TI - Analysis of HLA-G expression in tumor cell lines. PMID- 10371971 TI - Expression and regulation of HLA-G in human glioma cell lines. PMID- 10371973 TI - Transcriptional regulation of HLA-G. PMID- 10371972 TI - Preliminary study on the expression of HLA-G mRNA in normal placenta and placenta with RSA after immunotherapy. PMID- 10371974 TI - Induction of HLA-G-specific human CD8+ T cell lines by stimulation across a polymorphism of HLA-G. PMID- 10371975 TI - HLA-G expression on porcine endothelial cells protects partially against direct human NK cytotoxicity but not against ADCC. PMID- 10371976 TI - Choriocarcinoma cell line resistance to NK lysis mainly involves an HLA-G independent mechanism. PMID- 10371977 TI - Expression of HLA-E in transgenic mice. PMID- 10371978 TI - Structure of a human natural killer cell inhibitory receptor. PMID- 10371979 TI - First successful heart-lung transplantation for cystic fibrosis in Israel. PMID- 10371980 TI - Comparison of ELISA QuikScreen and PRA-STAT with CDC-AHG for PRA determination. PMID- 10371981 TI - Negative response to noninherited maternal antigens confirmed by ELISA. PMID- 10371982 TI - Initial 6-month experience with the new Israeli Organ Procurement Program: is there a penalty for expansion of the donor pool? PMID- 10371984 TI - Importance of the cardiothoracic ratio in the evaluation of cardiac transplant candidates. PMID- 10371983 TI - Differences in immunoactivation between heart and liver transplanted patients. PMID- 10371985 TI - Right atrial dilatation: major contributor to increased cardiothoracic ratio in cardiac transplant candidates. PMID- 10371986 TI - Peripheral vascular disease in pancreas allograft recipients. PMID- 10371987 TI - Graft versus host disease in a liver transplant patient with hepatitis B and hepatocellular carcinoma. PMID- 10371988 TI - Readmission to an intensive care unit following liver and kidney transplantation: a 50-month study. PMID- 10371989 TI - Solid tumors after liver transplantation. PMID- 10371990 TI - Primary sclerosing cholangitis and liver transplantation. PMID- 10371991 TI - The role of ERCP in biliary complications after liver transplantation. PMID- 10371992 TI - Pregnancy and liver transplantation. PMID- 10371993 TI - Reoperation after liver transplantation. PMID- 10371994 TI - Vascular complications post orthotopic liver transplantation. PMID- 10371995 TI - Employment is a misleading indicator for successful outcome after heart transplantation. PMID- 10371996 TI - Unexpected Wilms' tumor in a pediatric renal transplant recipient: suspected Denys-Drash syndrome. PMID- 10371997 TI - Israelis willingness to donate organs: result of a survey. PMID- 10371998 TI - Physicians' and nurses' attitudes and knowledge toward brain death. PMID- 10371999 TI - Lung transplants at Sheba Medical Center. PMID- 10372000 TI - Conversion from Sandimmune to Neoral in stable pediatric liver transplant recipients. PMID- 10372001 TI - Detection of anti-HLA antibodies by enzyme-linked immunosorbent assay, fluorescence activated cell sorter and microlymphocytotoxicity testing: a comparison of sensitivities and suggestions for standardization of ELISA. PMID- 10372006 TI - Surgical training in the 21st century. PMID- 10372007 TI - Guide for the management of breast lumps: litigious aspects. PMID- 10372008 TI - Soft-tissue images. Meckel's diverticulitis. PMID- 10372009 TI - Musculoskeletal images. Malignant change in long-standing enchondroma. PMID- 10372010 TI - Soft-tissue case 27. Magnetic resonance imaging: hepatic regenerative nodules. PMID- 10372011 TI - Musculoskeletal case 4. Idiopathic synovial osteochondromatosis. PMID- 10372012 TI - Transesophageal echocardiographic assessment in trauma and critical care. AB - Cardiac ultrasonography, in particular transesophageal echocardiography (TEE) provides high-quality real-time images of the beating heart and mediastinal structures. The addition of Doppler technology introduces a qualitative and quantitative assessment of blood flow in the heart and vascular structures. Because of its ease of insertion and ready accessibility, TEE has become an important tool in the routine management of critically ill patients, as a monitor in certain operative settings and in the aortic and cardiac evaluation of trauma patients. The rapid assessment of cardiac preload, contractility and valve function are invaluable in patients with acute hemodynamic decompensation in the intensive care unit as well as in the operating room. Because of its ease and portability, the TEE assessment of traumatic aortic injury after blunt chest trauma can be rapidly undertaken even in patients undergoing life-saving procedures. The role of TEE in the surgical and critical care setting will no doubt increase as more people become aware of its potential. PMID- 10372013 TI - Advocacy--answering old mail. Canadian Association of General Surgeons. AB - Since its inception in 1977, the Canadian Association of General Surgeons (CAGS) has struggled with its responsibility to represent general surgeons in practices across this country. The CAGS has tended to be mute in the presentation of many of its accomplishments, which have improved the role of specialists in community practice, training programs and the subspecialties of general surgery. With the forthcoming changes in direction for the Royal College of Physicians and Surgeons of Canada, based on a recent external survey, the CAGS has a golden opportunity to advocate for a clear identity, autonomous from the Royal College for the purposes of scientific meetings, continuing professional development, scientific and practice affiliation with other surgical specialty societies, and new developments with corporate sector support for advancements in science technology and education. Advocacy for general surgery must be stressed as the priority for the CAGS into the future. PMID- 10372014 TI - The urology work force in Ontario for the 21st century: feast or famine? AB - OBJECTIVE: To address the issues of work-force planning and modelling in the 21st century for the specialty of urology in the Province of Ontario. DESIGN: Data (from 1991 to 1995) regarding urology physician resources were gathered from Health Canada, the Royal College of Physicians and Surgeons of Canada, the Ontario Physician Human Resources Data Centre, the Canadian Post-M.D. Education Registry, the System for Health Area Resource Planning (SHARP) database, the Canadian Institute for Health Information and the National Physician Database. Specifically, the age and gender breakdown of currently active Ontario urologists, measures of urologist clinical activity (from Ontario Hospital Insurance Plan billings and questionnaires), inputs into and exits from the active urologist population were gathered, and estimates of future needs for urologist services, based on current population and demographic models, were made. A model to predict the balance between future needs for urology services and future supply of urologists was then created and validated against data drawn from the SHARP database. RESULTS: The model revealed that there will be a significant shortage of urologists in Ontario in the immediate and long-term future; by the year 2010 there will be a shortfall of 101 urologists in Ontario, or 51% of the total needed. CONCLUSIONS: Enlarging the urology training programs in Ontario would help to minimize the estimated shortfall. Systematic modelling of physician work-force needs for the future is necessary for the optimal allocation of health care resources. The methodology of the urology work-force model is generalizable to physician work-force planning for other specialty groups on a provincial or national basis. PMID- 10372015 TI - Doxorubicin-cisplatin chemotherapy for high-grade nonosteogenic sarcoma of bone. Comparison of treatment and control groups. AB - OBJECTIVE: To evaluate the role of chemotherapy with a combination of doxorubicin (adriamycin) and cisplatin in high-grade, nonosteogenic, non-Ewing's sarcoma (non OSA) of bone. DESIGN: A case series comparison with a literature-derived control group. SETTING: A university-affiliated tertiary care centre. PATIENTS: Thirty patients with a diagnosis of non-OSA. Of these, 8 had low-grade disease (grade 1 or 2) and 22 had high-grade disease (grade 3). Eleven of the 22 with high-grade disease had malignant fibrous histiocytoma. Seventeen patients with nonmetastatic high-grade non-OSA were compared with a literature cohort of 37 patients who met the eligibility criteria of nonmetastatic, high-grade non-OSA treated with surgery, with or without radiotherapy. The mean follow-up was 25.2 months. INTERVENTIONS: Eight patients with low-grade tumour underwent surgery alone; 22 patients with high-grade tumour underwent surgery and 6 courses of adriamycin (75 mg/m2 every 3 weeks) and cisplatin (100 mg/m2 every 3 weeks). MAIN OUTCOME MEASURES: Disease-free survival and overall survival in those with high-grade tumours treated with or without chemotherapy. RESULTS: Of 8 patients who had low grade tumours and underwent surgery alone, 3 had systemic relapse. Of the 22 having high-grade tumours, 4 did not receive chemotherapy because of age and comorbid conditions. Of the other 18, 13 received 3 courses of chemotherapy preoperatively and 3 courses postoperatively, 4 received all 6 courses postoperatively and 1 received all chemotherapy preoperatively to treat metastatic disease. In the 17-patient cohort used for comparison with the literature control group, disease-free survival was 57% at a mean follow-up of 25.6 months and overall survival was 57% at a mean follow-up of 30.1 months. In the control group, disease-free survival was 16% at a mean follow-up of 20.9 months and overall survival was 26% at a mean follow-up of 29.9 months. These differences are significant: p = 0.0000, chi 2 = 41.61 for disease-free survival and p = 0.0000, chi 2 = 46.49 for overall survival. CONCLUSIONS: The findings of this study support the use of adjuvant chemotherapy in patients with high-grade non-OSA, in whom malignant fibrous histiocytoma was the predominant histologic subtype. PMID- 10372016 TI - The hazards of vinyl glove ingestion in the mentally retarded patient with pica: new implications for surgical management. AB - OBJECTIVE: To report experience with the treatment of complications of vinyl glove ingestion in mentally retarded patients with pica. DESIGN: A retrospective case series. SETTING: Two university-affiliated hospitals. PATIENTS: Five mentally retarded patients, 4 with a history of pica, who were admitted for the management of complications resulting from the ingestion of vinyl gloves. MAIN OUTCOME MEASURES: Type of complication, treatment and operative outcome. FINDINGS: The patients ranged in age from 26 to 46 years. One patient died while awaiting surgical consultation of massive gastrointestinal bleeding from a large gastric ulcer caused by a vinyl glove bezoar (VGB). Four VGBs were removed surgically. Endoscopic removal was difficult or impossible because the gloves had become hardened and matted. CONCLUSIONS: VGB should be considered in institutionalized mentally retarded people with a history of pica when they present with gastrointestinal symptoms. VGBs should be removed directly by laparotomy, gastrotomy or enterotomy. Endoscopic removal is not recommended. PMID- 10372017 TI - Early experience with simulated trauma resuscitation. AB - Although trauma resuscitation is best taught through direct exposure with hands on experience, the opportunities for this type of teaching in Canada are limited by the relatively low incidence of serious injury and the consolidation of trauma care to a small number of centres. Simulators have been used extensively outside the health care environment and more recently have been used by anesthetists to simulate intraoperative crises. In this paper early experience using a realistic mannequin, controlled by a remote computer, that simulates a variety of physiologic and injury specific variables is presented. The resource implications of simulated resuscitation are reviewed, including one-time and operating costs. Simulated trauma resuscitation may be an educational alternative to "real-life" trauma resuscitation, but careful evaluation of the benefits and resource implications of this type of teaching through well-designed research studies will be important. PMID- 10372018 TI - The association between tamoxifen and the development of hepatocellular carcinoma: case report and literature review. AB - Tamoxifen has become one of the most widely used drugs in the treatment of breast cancer, and concerns about its long-term safety and efficacy are being raised. Investigations in rats have suggested an association between the administration of tamoxifen and the development of hepatocellular carcinoma. However, no studies to date have demonstrated an increased incidence of hepatocellular carcinoma in women treated with tamoxifen. In the case reported, a 56-year-old woman presented with hepatocellular tumours after 6 years of tamoxifen therapy for breast cancer. The patient had no other risk factors for the development of hepatocellular carcinoma. She underwent successful resection of the lesions, and subsequent pathological studies confirmed hepatocellular carcinoma with a trabecular growth pattern similar to the histologic pattern seen in tamoxifen-induced hepatocellular carcinoma occurring in rat models. PMID- 10372019 TI - Post-traumatic pancreatitis with associated aneurysm of the splenic artery: report of 2 cases and review of the literature. AB - In patients with acute pancreatitis, profuse gastrointestinal bleeding is associated with a high death rate. The cause of such bleeding must be evaluated and the bleeding controlled urgently. Aneurysm formation is usually the cause of the bleeding. Angiography is needed to make a definitive diagnosis and the bleeding site should be controlled by angiographic embolization if possible. If this fails, aneurysm resection is necessary. Two patients are described. Both had aneurysms of the splenic artery, presenting as massive gastrointestinal bleeding in one patient and bleeding into an associated pseudocyst in the other. They required surgical repair, which was successful in both cases. PMID- 10372020 TI - Simultaneous pyloric and colonic obstruction associated with hiatus hernia in a weightlifter: a case report. AB - Hiatus hernia is usually attributed to conditions that cause a chronic increase in intra-abdominal pressure such as multiple pregnancies and obesity. A 30-year old man, a weightlifter, had a massive hiatus hernia causing both high and low gastrointestinal obstruction but no involvement of the gastroesophageal junction or fundus. The onset of the obstruction is attributed to an extreme increase in intra-abdominal pressure caused by the action of lifting weights. PMID- 10372021 TI - The human genome project: a dream becoming a reality. PMID- 10372022 TI - Early and late outcomes of young patients after carotid endarterectomy. AB - BACKGROUND AND PURPOSE: Carotid atherosclerotic disease in young adults is uncommon but may be more virulent and diffuse than in older patients. Although few studies have well established that carotid endarterectomy (CEA) is of benefit for high-grade asymptomatic lesions and for moderate- and high-grade symptomatic lesions, the safety and durability of CEA in the young remain controversial. The aim of this study was to compare CEA outcome in young people with outcome in an older control group. METHODS: Thirty-five patients up to 50 years old (mean 46.5 +/- 0.5 years) and undergoing 42 CEAs were compared with a randomly selected group of 50 patients more than 60 years old (mean 68.7 +/- 0.4 years) and undergoing 55 CEAs during the same period. Data were obtained on demographics, atherosclerotic risk factors, indications for surgery, perioperative outcome, recurrent stenosis and symptoms, late stroke, and survival. RESULTS: Smoking (P < .001), alcohol consumption (P < .001), and lower levels of high-density lipoprotein cholesterol (P = .02) were more prevalent in the young patients, who were also more likely to be symptomatic at presentation (P < .001) with a higher incidence of stroke (P = .01) and contralateral carotid occlusion (P = .04). The perioperative stroke risk and mortality rates were nil for the young group. During a mean follow-up of 47 +/- 40 months, there were no significant differences between the 2 groups in survival, symptom recurrence, stenosis recurrence, and reoperation rates. Young patients had a higher incidence of contralateral disease requiring surgery (P = .04). CONCLUSIONS: These findings show that CEA may be performed in young adults with an excellent perioperative outcome; recurrence, late stroke, and survival rates do not differ significantly from those observed among their older counterparts. PMID- 10372023 TI - Temporal activity of plasminogen activators and matrix metalloproteinases during cutaneous wound repair. AB - BACKGROUND: Response to tissue injury begins with the deposition of a fibrin-rich clot or the provisional matrix. The provisional matrix consists of plasma-borne matrix molecules that serve as scaffolding for the ensuing migration of cells. During wound repair multiple cell types must migrate through the clot-matrix scaffolding. The migration of these cells through the matrix is dependent on the activity of the fibrinolytic and proteolytic systems, which include the plasminogen activator (PA) system and matrix metalloproteinases (MMP). The aim of this study was to better understand the temporal activity of these enzymes during normal wound repair. METHODS: We used the murine excisional wound model and extracted proteins under nonreducing conditions. With use of gelatin and casein zymography, we determined the activity of the MMPs during the course of wound repair. In addition, we quantified the activity of MMP-2 and MMP-9 by a standardized assay. Plasminogen zymograms were used to detect urokinase PA and tissue PA activity. Western blots were used to detect the natural inhibitor of PAs, plasminogen activator inhibitor type 1. RESULTS: Our results demonstrate the temporal activity of MMP-2, MMP-3, MMP-7, and MMP-9 during the course of normal dermal repair. The activity of urokinase PA and tissue PA were also determined; it preceded the activity of the MMPs. CONCLUSIONS: We demonstrate the temporal activity of the 2 protease families, MMPs and PAs, in the normal process of cutaneous wound healing. PMID- 10372024 TI - Heat-shock protein 72/73 and impaired wound healing in diabetic and hypercortisolemic states. AB - BACKGROUND: Impaired wound healing is a well-documented phenomenon in experimental and clinical diabetes. Emerging evidence favors the involvement of glucocorticoids (GCs) in the pathogenesis of this diabetic complication. Recent data indicated that a heat-shock protein (HSP) with a molecular weight of about 70 kd is expressed in wound healing and it is under the control of the hypothalamic-pituitary-adrenal axis. In view of these findings, the current study was designed to examine the influence of diabetes and the hypercortisolemic state on the expression of HSP 72/73 during wound healing. METHODS: Induction of diabetes was achieved by the intravenous injection of streptozotocin at a dose of 55 mg/kg. Subcutaneously implanted polyvinyl alcohol (PVA) sponges were used as a wound healing model. Control and diabetic animals received, respectively, subcutaneous 30-day timed-release pellets of GC (200 mg) and RU 486 (25 mg). Corresponding animals received placebo pellets. Expression of HSP 72/73 within the PVA sponges was assayed with use of Western blotting and immunohistochemical techniques. RESULTS: GCs caused a Cushing-like syndrome with weight loss and adrenal atrophy. A pronounced accumulation of constitutive HSP 72/73 was observed in the cytoplasm of various cell types including fibroblasts, macrophages, and endothelium of nondiabetic controls. The PVA sponge contents of HSP 72/73 were decreased as a function of diabetes. A similar phenomenon was seen in control animals receiving high doses of GCs. Partial normalization of the associated hyperglycemic and hypercortisolemic states of diabetes with insulin (hyperglycemia) and the GC receptor block RU 486 (hypercortisolemia) ameliorated the diabetes-related decrease in PVA sponge contents of HSP 72/73. CONCLUSIONS: The current study provides evidence that both diabetes and the hypercortisolemic state are associated with a reduction in PVA sponge content of HSP 72/73. An amelioration of these changes was achieved by the institution of RU 486 therapy. Although our data may point to the possibility that the diabetes-related decrease in HSP 72/73 is mediated at least in part by GCs, a confirmation regarding this premise awaits further investigation. PMID- 10372025 TI - Deletion variant of hepatocyte growth factor prolongs allograft survival after liver transplantation in rats. AB - BACKGROUND: Despite continued progress in the development of immunosuppressive agents, allograft rejection remains an important cause of morbidity and mortality after liver transplantation. We examined the effect of the deletion variant of hepatocyte growth factor (dHGF) on allograft rejection after liver transplantation. METHODS: Male Dark Agouti rats (RT1a) were selected as donors and male Lewis rats (RT1l) as recipients for a rejection model. The recipients were divided into 2 groups after orthotopic liver transplantation (OLTx): in the dHGF group dHGF was given intravenously twice a day (1 mg/kg/day) after OLTx, whereas in the control group vehicle buffer was given intravenously daily twice after OLTx. The survival period, serum chemistry studies, and histopathologic findings were then compared between the 2 groups. RESULTS: The mean survival period after OLTx in the dHGF group was significantly longer than that in the control group (21.4 +/- 1.3 days vs 11.8 +/- 0.4 days, P < .001). On the 10th posttransplant day the serum albumin level significantly improved in the dHGF group (P < .01), and the serum total bilirubin and aspartate aminotransferase levels were significantly lower in the dHGF group (P < .01 and P < .05, respectively). On the 10th posttransplant day a histologic examination revealed no apparent difference in the severity of rejection between the 2 groups. The number of proliferating cell nuclear antigen-positive hepatocytes in the dHGF group significantly increased (P < .01), whereas the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling-positive hepatocytes were significantly reduced in the dHGF group (P < .01) in comparison with those in the control group. CONCLUSION: dHGF has an antiapoptotic property as well as a proliferative and protective effect on hepatocytes under allograft rejection. dHGF might serve as a novel agent for reducing the harmful effects of hepatic allograft rejection in rats. PMID- 10372026 TI - Health status improvement after surgical correction of primary hyperparathyroidism in patients with high and low preoperative calcium levels. AB - BACKGROUND: We conducted a prospective cohort study to determine whether there are differences in functional health status between patients with low (< 10.9 mg/dL) and high (> or = 10.9 mg/dL) serum calcium levels before surgical correction of primary hyperparathyroidism (HPT) and to compare changes in health status after correction of primary HPT. METHODS: The SF-36 Health Survey, which provides demographic and condition-specific information, was used to obtain information on patients with primary HPT seen in a university hospital endocrine surgery clinic over a 4-year period before operation and again 2 months and 6 months after operation. RESULTS: A total of 155 patients were studied; 86 had calcium levels < 10.9 mg/dL (normal < 10.5 mg/dL) and 69 had serum calcium levels > or = 10.9 mg/dL (range 10.9 to 13.4 mg/dL). One hundred four patients completed 6-month reports, 55 with low calcium levels and 49 with high calcium levels. Both high and low calcium groups showed marked and virtually identical impairment of functional health status. Both groups showed marked improvement in health status at 2 months and additional improvement at 6 months, returning to normal or near normal in 6 of 8 SF-36 domains. CONCLUSIONS: Patients with primary hyperparathyroidism have significant functional health status impairment independent of the level of serum calcium. Dramatic improvement is seen after surgical correction. Referral for surgical treatment of primary HPT should not be delayed until serum calcium is elevated, as recommended in the 1990 National Institutes of Health consensus statement. PMID- 10372027 TI - How competitive forces mold strategy in academic surgery. PMID- 10372028 TI - Advice from a vendor: play the trump card. PMID- 10372029 TI - Talk the game, then play the game. PMID- 10372030 TI - Aortic aneurysm associated with systemic lupus erythematosus. PMID- 10372031 TI - Don't be shocked: implantable defibrillators during surgery. PMID- 10372032 TI - Practical issues in medication compliance. PMID- 10372034 TI - Nonadherence to immunosuppressive medications: a pilot survey of members of the transplant recipients international organization. PMID- 10372033 TI - Patient compliance in transplantation: a report on the perceptions of transplant clinicians. PMID- 10372035 TI - Health-care professional perceptions of compliance behaviors in the prerenal and postrenal transplant patient. PMID- 10372036 TI - Lifestyle assessment and heart transplant evaluation. PMID- 10372037 TI - Noncompliance in living-related donor renal transplantation: the United Network of Organ Sharing experience. PMID- 10372038 TI - Late renal allograft loss: noncompliance masquerading as chronic rejection. PMID- 10372039 TI - Cross-cultural issues in transplant compliance. PMID- 10372040 TI - Medication adherence and the transplant recipient: helping patients at each stage of change. PMID- 10372041 TI - A medical anthropologist's view on posttransplant compliance: the underground economy of medical survival. PMID- 10372042 TI - United Resource Networks: how compliance issues affect a managed care transplant network. PMID- 10372043 TI - The impact of compliance on drug development. PMID- 10372044 TI - Impact of compliance on drug development: an FDA perspective. PMID- 10372045 TI - Increased compliance through targeted behavioral interventions. PMID- 10372046 TI - Theories of group intervention. PMID- 10372048 TI - Pharmacoeconomic evaluations of immunosuppressive agents. PMID- 10372047 TI - Addiction relapse prevention in solid-organ transplantation. PMID- 10372049 TI - Role of the transplant financial coordinator and its effect on recipient compliance. PMID- 10372050 TI - Posttransplant quality of life issues: depression-related noncompliance in cardiac transplant patients. PMID- 10372051 TI - Pediatric renal transplantation: back to school issues. PMID- 10372052 TI - Liquid medication dispensing and dose monitoring: the CycloTech Cyclosporine Oral Solution Dispenser. AB - This liquid medication dispenser offers an easy, convenient means for accurate dispensing of medication. The ability of the device to store dose size, time to next dose, remaining available doses, and doses dispensed may allow for future analysis of patient behavior and improve compliance. PMID- 10372053 TI - Solid medication dispensing: the polypharm experience and lessons from other disease states. PMID- 10372054 TI - An individualized teaching and self-medication program to promote compliance in newly transplanted pediatric patients. PMID- 10372055 TI - Peer-to-peer transplant mentor program: the San Diego experience. PMID- 10372056 TI - The Arizona experience: empowering patients and families. PMID- 10372057 TI - Methodological issues in transplant compliance research. PMID- 10372058 TI - A tool to identify risk factors for noncompliance in the adult renal transplant recipient. PMID- 10372059 TI - Patient compliance with cyclosporine regimen post solid organ transplantation. PMID- 10372060 TI - Stimulus processing by type II hair cells in the mouse utricle. AB - In type II and neonatal hair cells in the mouse utricle, the receptor potentials evoked by low-frequency sinusoidal deflections of the hair bundle are attenuated by adaptation of the mechanoelectrical transduction current and the voltage dependent activation of a large potassium (K)-selective outwardly rectifying conductance, gDR. These processes may contribute to high-pass filtering of the responses of some utricular afferents to sinusoidal linear accelerations below 2 Hz. Depolarizing receptor potentials are more attenuated by gDR than are hyperpolarizing receptor potentials. It may therefore reduce nonlinear distortion introduced by mechanoelectrical transduction, which generates larger depolarizing currents than hyperpolarizing currents. The discharge properties of utricular afferents vary according to whether they innervate the striolar or extrastriolar zones of the sensory epithelium. Regional variation in hair-cell properties is likely to contribute. Preliminary results suggest that the outwardly rectifying K conductances of type II cells are slower and larger in the striola than in the extrastriola, consistent with regional variation in the relative numbers of delayed rectifier and A-current K channels. PMID- 10372061 TI - The planes of the utricular and saccular maculae of the guinea pig. AB - To establish a link between otolith anatomy and function it is necessary to know the regions of the utricular and saccular maculae, which are stimulated by any arbitrary linear acceleration stimulus. That requires accurate information about the location and orientation of the spatially extended maculae in head-fixed coordinates and referred to head-fixed landmarks (such as Reid's line). New data showing the location of the otolithic maculae in the guinea pig with respect to head-fixed stereotaxic coordinates are presented. Guinea pigs were perfused with Karnovsky's fixative and the maculae were exposed while the head was held in a guinea pig stereotaxic device. An electrolytically sharpened fine wire held in a calibrated micromanipulator was touched to points all over the surface of each macula under visual observation with the aid of a high-power operating microscope. The x, y, z coordinates of these points were plotted using a three dimensional plotting program. Both maculae have pronounced curvature so that dorsoventral shear forces will stimulate regions of both the utricular and saccular maculae. PMID- 10372062 TI - A review of otolith pathways to brainstem and cerebellum. AB - Our knowledge of otolith pathways is developing rapidly, but is still far from complete. Primary afferents from the sacculus and utricle terminate mainly in the lateral, inferior and caudal superior vestibular nuclei, and the ventral cerebellum, in particular the nodulus. Otolith signals descend via reticulo- and vestibulospinal pathways in the spinal cord to influence neck motoneurons and ascending proprioceptive afferents. Utricular information can reach the extraocular eye muscles via mono-, di-, and multisynaptic pathways, but saccular afferents probably only by multisynaptic pathways. The otolith signals are relayed from the vestibular nuclei, medullary reticular formation, inferior olive, and lateral reticular nucleus to sagittal zones in the caudal cerebellar vermis (nodulus and uvula), and influence the deep cerebellar nuclei. The graviceptive information could be channeled by the cerebellar efferents back to the vestibular and inferior olive complex, or fed into ascending pathways that would innervate the mescencephalon, the thalamus, and cerebral cortex. PMID- 10372063 TI - Signal processing related to the vestibulo-ocular reflex during combined angular rotation and linear translation of the head. AB - The contributions of vestibular nerve afferents and central vestibular pathways to the angular (AVOR) and linear (LVOR) vestibulo-ocular reflex were studied in squirrel monkeys during fixation of near and far targets. Irregular vestibular afferents did not appear to be necessary for the LVOR, since when they were selectively silenced with galvanic currents the LVOR was essentially unaffected during both far- and near-target viewing. The linear translation signals generated by secondary AVOR neurons in the vestibular nuclei were, on average, in phase with head velocity, inversely related to viewing distance, and were nearly as strong as AVOR-related signals. We suggest that spatial-temporal transformation of linear head translation signals to angular eye velocity commands is accomplished primarily by the addition of viewing distance multiplied, centrally integrated, otolith regular afferent signals to angular VOR pathways. PMID- 10372064 TI - Otolith processing in the deep cerebellar nuclei. AB - To investigate the otolith contribution to the responses of "vestibular only" neurons in the rostral fastigial nucleus (FN), single-unit activity was recorded in the alert monkey with the head fixed during static and dynamic stimulation (+/ 15 deg, 0.06-1.4 Hz) around an earth-fixed horizontal axis. Head orientation could be altered allowing for roll, pitch, and intermediate planes of orientation. For the vast majority of neurons a response vector orientation (RVO) with an optimal response and a null-response at a head orientation 90 deg apart could be determined. Presumably more than 30% of the vestibular only neurons had an otolith input, as indicated by responses to static tilt, head-position-related activity, large phase changes (> 100 deg) of neuronal activity between 0.06 and 1.4 Hz, changes of the RVO at different frequencies and complex responses (spatio temporal convergence). Thus, neurons in FN reflecting an otolith or a combined canal-otolith input are much more common than up to now thought. Vestibular-only neurons are most likely involved in vestibulospinal mechanisms. Their precise functional role has yet to be determined. PMID- 10372066 TI - Characteristics of the VOR in response to linear acceleration. AB - The primate linear VOR (LVOR) includes two forms. First, eye-movement responses to translation [e.g., horizontal responses to interaural (i.a.) motion] help maintain binocular fixation on targets, and therefore a stable bifoveal image. The translational LVOR is strongly modulated by fixation distance, and operates with high-pass dynamics (> 1 Hz). Second, other LVOR responses occur that cannot be compensatory for translation and instead seem compensatory for head tilt. This reflects an otolith response ambiguity--that is, an inability to distinguish head translation from head tilt relative to gravity. Thus, ocular torsion is appropriately compensatory for head roll-tilt, but also occurs during IA translation, since both stimuli entail IA acceleration. Unlike the IA-horizontal response, IA torsion behaves with low-pass dynamics (with respect to "tilt"), and is uninfluenced by fixation distance. Interestingly, roll-tilt, like IA translation, also produces both horizontal (a translational reflex) and torsional (a tilt reflex) responses, further emphasizing the ambiguity problem. Early data from subjects following unilateral labyrinthectomy, which demonstrates a general immediate decline in translational LVOR responses, are also presented, followed by only modest recovery over several months. Interestingly, the usual high-pass dynamics of these reflexes shift to an even higher cutoff. Both eyes respond roughly equally, suggesting that unilateral otolith input generates a binocularly symmetric LVOR. PMID- 10372065 TI - Control of spatial orientation of the angular vestibulo-ocular reflex by the nodulus and uvula of the vestibulocerebellum. AB - Eye velocity produced by the angular vestibulo-ocular reflex (aVOR) tends to align with the summed vector of gravity and other linear accelerations [gravito inertial acceleration (GIA)]. Defined as "spatial orientation of the aVOR," we propose that it is controlled by the nodulus and uvula of the vestibulocerebellum. Here, electrical stimulation, injections of the GABAA agonist, muscimol, and single-cell recordings were utilized to investigate this spatial orientation. Stimulation, injection, and recording sites in the nodulus were determined in vivo by MRI and verified in histological sections. MRI proved to be a sensitive, reliable way to localize electrode placements. Electrical stimulation at sites in the nodulus and sublobule d of the uvula produced nystagmus whose slow-phase eye-velocity vectors were either head centric or spatially invariant. When head centric, the eye velocity vector remained within +/- 45 degrees of the vector obtained with the animal upright, regardless of head position with respect to gravity. When spatially oriented, the vector remained relatively constant in space in one on-side position, with respect to the vector determined with the animal upright. A majority of induced movements from the nodulus were spatially oriented. Spatially oriented movements were generally followed by after-nystagmus, which had the characteristics of optokinetic after nystagmus (OKAN), including orientation to the GIA. After muscimol injections, horizontal-to-vertical cross-coupling was lost or reduced during OKAN in tilted positions. This supports the hypothesis that the nodulus mediates yaw-to-vertical or roll cross-coupling. The injections also shortened the yaw-axis time constant and produced contralateral horizontal spontaneous nystagmus, whose velocity varied as a function of head position with regard to gravity. Nodulus units were tested with static head tilt, sinusoidal oscillation around a spatial horizontal axis with the head in different orientations relative to the pitching plane, and off-vertical axis rotation (OVAR). The direction of the response vectors of the otolith-recipient units in the nodulus, determined from static and/or dynamic head tilts, were confirmed by OVAR. These vector directions lay close to the planes of the vertical canals in 7/10 units; many units also had convergent input from the vertical canals. It is postulated that the orientation properties of the aVOR result from a transfer of otolith input regarding head tilt along canal planes to canal-related zones of the nodulus. In turn, Purkinje cells in these zones project to vestibular nuclei neurons to control eye velocity around axes normal to these same canal planes. PMID- 10372067 TI - Functional organization of primate translational vestibulo-ocular reflexes and effects of unilateral labyrinthectomy. AB - Translational vestibulo-ocular reflexes (trVORs) are characterized by distinct spatio-temporal properties and sensitivities that are proportional to the inverse of viewing distance. Anodal (inhibitory) labyrinthine stimulation (100 microA, < 2 s) during motion decreased the high-pass filtered dynamics, as well as horizontal trVOR sensitivity and its dependence on viewing distance. Cathodal (excitatory) currents had opposite effects. Translational VORs were also affected after unilateral labyrinthectomy. Animals lost their ability to modulate trVOR sensitivity as a function of viewing distance acutely after the lesion. These deficits partially recovered over time, albeit a significant reduction in trVOR sensitivity as a function of viewing distance remained in compensated animals. During fore-aft motion, the effects of unilateral labyrinthectomy were more dramatic. Both acute and compensated animals permanently lost their ability to modulate fore-aft trVOR responses as a function of target eccentricity. These results suggest that (1) the dynamics and viewing distance-dependent properties of the trVORs are very sensitive to changes in the resting firing rate of vestibular afferents and, consequently, vestibular nuclei neurons; (2) the most irregularly firing primary otolith afferents that are most sensitive to labyrinthine electrical stimulation might contribute to reflex dynamics and sensitivity; (3) inputs from both labyrinths are necessary for the generation of the translational VORs. PMID- 10372068 TI - Inertial processing of vestibulo-ocular signals. AB - New evidence for a central resolution of gravito-inertial signals has been recently obtained by analyzing the properties of the vestibulo-ocular reflex (VOR) in response to combined lateral translations and roll tilts of the head. It is found that the VOR generates robust compensatory horizontal eye movements independent of whether or not the interaural translatory acceleration component is canceled out by a gravitational acceleration component due to simultaneous roll-tilt. This response property of the VOR depends on functional semicircular canals, suggesting that the brain uses both otolith and semicircular canal signals to estimate head motion relative to inertial space. Vestibular information about dynamic head attitude relative to gravity is the basis for computing head (and body) angular velocity relative to inertial space. Available evidence suggests that the inertial vestibular system controls both head attitude and velocity with respect to a gravity-centered reference frame. The basic computational principles underlying the inertial processing of otolith and semicircular canal afferent signals are outlined. PMID- 10372069 TI - Cross-striolar and commissural inhibition in the otolith system. AB - Neural connections from the saccular and utricular nerves to the ipsilateral vestibular neurons and the commissural effects were studied by using intracellular recordings of excitatory (E) and inhibitory (I) postsynaptic potentials (PSPs) in vestibular neurons of cats after focal stimulation of the saccular and the utricular maculae. Neural circuits from the maculae to vestibular neurons, termed cross-striolar inhibition, may provide a mechanism for increasing the sensitivity to linear acceleration and tilt of the head. It was examined whether secondary vestibular neurons activated by an ipsilateral otolith organ received a commissural inhibition from a contralateral otolith organ that occupied the same geometric plane. Results suggest that utricular-activated vestibular neurons receiving commissural inhibition may provide a mechanism for increasing the sensitivity to horizontal linear acceleration and tilt of the head. The commissural inhibition of the saccular system was much weaker than that of the utricular system. PMID- 10372070 TI - Human ocular counterrolling during roll-tilt and centrifugation. AB - To test a hypothesis about how otoliths resolve roll-tilts from translations, we measured human ocular torsion position [ocular counterrolling (OCR)] to maintained linear acceleration stimuli. All subjects (n = 8) were tested in two conditions where the same magnitude of shear along an interaural axis was generated in one of two ways: either by roll-tilt on a tilt-chair in a 1-g environment, or by centripetal linear acceleration during constant velocity rotation 1 m from the axis of rotation on a fixed-chair human centrifuge. The interaural shear to the otoliths was the same for these two conditions, but the dorsoventral shear was different and for all eight subjects the OCR on the centrifuge was significantly greater than the torsion on the tilt-chair, although the resultant angle was in fact smaller on the centrifuge than on the tilt-chair. The results confirm that dorsoventral shear is important for determining OCR. The otoliths may resolve potential stimulus ambiguities between tilts and translations by virtue of the different patterns of interaural and dorsoventral shear that these stimuli generate. PMID- 10372072 TI - Clinical testing of otolith function. AB - The subjective visual horizontal and vestibular-evoked myogenic potentials are simple, robust, and reproducible tests of otolith dysfunction that can prove clinically useful diagnostic information in patients with vertigo and other balance disorders. While they appear to have high specificity for unilateral otolith dysfunction, further clinical research will be required to establish their sensitivity. PMID- 10372073 TI - Directional abnormalities of vestibular and optokinetic responses in cerebellar disease. AB - Directional abnormalities of vestibular and optokinetic responses in patients with cerebellar degeneration are reported. Three-axis magnetic search-coil recordings of the eye and head were performed in eight cerebellar patients. Among these patients, examples of directional cross-coupling were found during (1) high frequency, high-acceleration head thrusts; (2) constant-velocity chair rotations with the head fixed; (3) constant-velocity optokinetic stimulation; and (4) following repetitive head shaking. Cross-coupling during horizontal head thrusts consisted of an inappropriate upward eye-velocity component. In some patients, sustained constant-velocity yaw-axis chair rotations produced a mixed horizontal torsional nystagmus and/or an increase in the baseline vertical slow-phase velocity. Following horizontal head shaking, some patients showed an increase in the slow-phase velocity of their downbeat nystagmus. These various forms of cross coupling did not necessarily occur to the same degree in a given patient; this suggests that different mechanisms may be responsible. It is suggested that cross coupling during head thrusts may reflect a loss of calibration of brainstem connections involved in the direct vestibular pathways, perhaps due to dysfunction of the flocculus. Cross-coupling during constant-velocity rotations and following head shaking may result from a misorientation of the angular eye velocity vector in the velocity-storage system. Finally, responses to horizontal optokinetic stimulation included an inappropriate torsional component in some patients. This suggests that the underlying organization of horizontal optokinetic tracking is in labyrinthine coordinates. The findings are also consistent with prior animal-lesion studies that have shown a role for the vestibulocerebellum in the control of the direction of the VOR. PMID- 10372071 TI - Canal and otolith afferent activity underlying eye velocity responses to pitching while rotating. AB - Pitching the head while rotating (PWR) combines periodic activation of the semicircular canals and the otoliths to generate pitch and roll eye deviations and continuous horizontal nystagmus. Monkeys were tested after individual pairs of semicircular canals were plugged and single units were recorded in the vestibular nerve while the animals were sinusoidally pitched 20-40 deg about a spatial horizontal axis with 5- and 16-s periods and simultaneously rotated about a spatial vertical axis at 30-120 deg/s. As previously shown, the steady-state horizontal response disappeared after plugging the vertical semicircular canals, but was maintained when the lateral canals were plugged. When the left anterior and right posterior canal (LARP) pair was left intact, the steady-state response depended on the axis about which the pitching took place. When the axis was normal to the LARP plane, there was no steady-state response. When the pitching axis was perpendicular to the LARP normal, the response was maximal. Firing rates of otolith units were approximately in phase with pitch position, and the addition of rotation about a vertical axis did not change the response. Lateral canal units did not have a steady-state modulation during pitch or constant velocity rotation. During PWR, they oscillated at twice the pitch frequency. This corresponded to the frequency at which the canal was maximally activated as it aligned with the plane of rotation. The amplitude of modulation increased proportionally to rotational velocity, but the phase remained the same. These characteristics were unchanged during roll while rotating (RWR), which induces little continuous nystagmus. Anterior and posterior canal units were maximally excited near pitch-velocity maxima and minima, respectively, during pure pitching. During PWR, however, the phases of both components simultaneously shifted toward each other and toward being in phase with otolith units. The peak excitation tended toward a forward-pitch position when the rotation was to the ipsilateral side, and toward a backward pitch position when the rotation was to the contralateral side. With 120-deg/s rotation during a 16-s pitch period, the phase difference between anterior and posterior canal units was as small as 17 deg. These data support the postulate that the correlation between vertical canal and otolith units is the critical factor in generating continuous unidirectional horizontal nystagmus during PWR. PMID- 10372074 TI - Assessing otolith function by the subjective visual vertical. AB - The effects of peripheral vestibular diseases on the subjective visual vertical (SVV) are resumed and provide the basis for some insights into the otolith pathophysiology. With a normal range of 0 +/- 2 deg (when measured in an upright body position), the SVV was shifted by 11 +/- 6 deg toward the ipsilateral ear in 40 patients following an acute unilateral vestibular deafferentiation (UVD), but in the opposite direction in 9 of 52 patients after stapes surgery. These opposite effects suggest a push-pull mechanism of the pairs of otolith organs with respect to the SVV. The dissociation between the SVV and the perception of body position indicates influences by unconscious reflexive mechanisms such as ocular cyclotorsion on the SVV. In chronic UVD patients, lateral shifts of the subjects during constant angular velocity rotation into various eccentric positions (+/- 16 cm) revealed a shift of the "center of graviception" close to the remaining intact contralateral inner ear. To date, this seems to be the most consistent test for clinical identification of a chronic compensated unilateral loss of otolith function. The findings regarding asymmetries in otolithic sensitivity to medially and laterally directed roll-tilts remain controversial, probably mainly because of influences of extravestibular cues. PMID- 10372075 TI - Horizontal linear vestibulo-ocular reflex testing in patients with peripheral vestibular disorders. AB - Horizontal eye movements in response to lateral head translation [linear vestibulo-ocular reflex (LVOR)] in normal subjects and in patients with bilateral vestibular failure (n = 14), unilateral vestibular nerve section (n = 9), and benign positional vertigo (n = 14), were studied. LVORs were elicited in darkness by step acceleration (0.24 g) of the whole body along the interaural axis. RESULTS AND CONCLUSIONS: (1) In patients with bilateral vestibular failure, LVORs were either absent or abnormal with asymmetries, diminished velocities, and prolonged latencies. Measurements of dynamic visual acuity during linear self motion showed decreased performance in patients at 1.0 and 1.5 Hz, which correlated with absent or delayed LVORs. These findings demonstrate the functional role of LVORs for dynamic visual acuity. (2) Early after vestibular nerve section, LVORs were diminished or absent with head acceleration toward the operated ear and normal in the opposite direction. After 6-10 weeks, responses were symmetrical again. Thus, a single utricle appears to be polarized with respect to the LVOR early after unilateral vestibular loss generating mostly contraversive responses. (3) Patients with benign positional vertigo showed mostly normal LVORs, which can be explained by minor utricular damage or central compensation of a chronic unilateral deficit. PMID- 10372076 TI - Vestibular-pursuit interactions: gaze-velocity and target-velocity signals in the monkey frontal eye fields. AB - Visual information about a moving object is obtained by accurate tracking with the eyes using the smooth pursuit system, which must interact with the vestibular system during head movement. Such pursuit-vestibular interactions require calculation of gaze (i.e., eye in space) in order to match eye velocity in space to actual target velocity, using vestibular, retinal-image velocity, and eye velocity information. To understand the role the frontal eye fields (FEFs) play in pursuit-vestibular interactions, we examined responses of pursuit-related neurons near the arcuate sulcus in head-stabilized monkeys during visual tracking tasks that dissociate eye movement in the orbit from that in space. The activity of the majority of pursuit-related neurons was related to gaze velocity. They also responded to passive body rotation in complete darkness. When the monkeys fixated the stationary target, similar modulation was observed, reflecting the velocity signal of a second test target. Muscimol infusion into the FEF pursuit areas severely impaired smooth gaze tracking. These results suggest that the region near the arcuate sulcus coordinates its various inputs to provide signals for target velocity in space and accurate gaze-velocity command during pursuit vestibular interactions. PMID- 10372077 TI - Short-latency visual stabilization mechanisms that help to compensate for translational disturbances of gaze. AB - Recent studies in primates have revealed short-latency visual tracking mechanisms that help to stabilize the eyes during translational disturbances of the observer, and so operate as backups to otolith-mediated vestibulo-ocular reflexes. One such mechanism generates version eye movements to help stabilize gaze when the moving observer looks off to one side, utilizing binocular disparity to help single out the images in the plane of fixation (ocular following). Two others generate vergence eye movements to help maintain binocular alignment on objects that lie ahead: one responds to the radial patterns of optic flow (radial-flow vergence) and the other to the changes in binocular parallax (disparity vergence). Accumulating evidence suggests that, despite their short latency, all are mediated by the medial superior temporal area of cortex. PMID- 10372078 TI - Linear vestibular self-motion signals in monkey medial superior temporal area. AB - The present study was aimed at investigating the sensitivity to linear vestibular stimulation of neurons in the medial superior temporal area (MST) of the macaque monkey. Two monkeys were moved on a parallel swing while single-unit activity was recorded. About one-half of the cells (28/51) responded in the dark either to forward motion (n = 10), or to backward motion (n = 11), or to both (n = 7). Twenty cells responding to vestibular stimulation in darkness were also tested for their responses to optic flow stimulation simulating forward and backward self-motion. Forty-five percent (9/20) of them preferred the same self-motion directions, that is, combined visual and vestibular signals in a synergistic manner. Thirty percent (6/20) of the cells were not responsive to visual stimulation alone. The remaining 25% (5/20) preferred directions that were antialigned. Our results provide strong evidence that neurons in the MST area are at least in part involved in the processing of self-motion. PMID- 10372079 TI - The contributions of vestibular signals to the representations of space in the posterior parietal cortex. AB - Vestibular signals play an important role in spatial orientation, perception of object location, and control of self-motion. Prior physiological research on vestibular information processing has focused on brainstem mechanisms; relatively little is known about the processing of vestibular information at the level of the cerebral cortex. Recent electrophysiological experiments examining the use of vestibular canal signals in two different perceptual tasks are described: computation of self motion and localization of visual stimuli in a world-centered reference frame. These two perceptual functions are mediated by different parts of the posterior parietal cortex, the former in the dorsal aspect of the medial superior temporal area (MSTd) and the latter in area 7a. PMID- 10372080 TI - The vestibular cortex. Its locations, functions, and disorders. AB - Evidence is presented that the multisensory parieto-insular cortex is the human homologue of the parieto-insular vestibular cortex (PIVC) in the monkey and is involved in the perception of verticality and self-motion. Acute lesions (patients with middle cerebral artery infarctions) of this area caused contraversive tilts of perceived vertical, body lateropulsion, and, rarely, rotational vertigo. Brain activation studies using positron emission tomography or functional magnetic resonance tomography showed that PIVC was activated by caloric irrigation of the ears or by galvanic stimulation of the mastoid. This indicates that PIVC receives input from both the semicircular canals and otoliths. PIVC was also activated during small-field optokinetic stimulation, but not when the nystagmus was suppressed by fixation. Activation of vestibular cortex areas, visual motion-sensitive areas, and ocular motor areas exhibited a significant right-hemispheric dominance. The vestibular cortex intimately interacts with the visual cortex to match the two 3-D orientation maps (perception of verticality, room-tilt illusion) and mediates self-motion perception by means of a reciprocal inhibitory visual-vestibular interaction. This mechanism of an inhibitory interaction allows a shift of the dominant sensorial weight during self-motion perception from one sensory modality (visual or vestibular) to the other, depending on which mode of stimulation prevails: body acceleration (vestibular input) or constant velocity motion (visual input). PMID- 10372081 TI - Cortical areas activated by bilateral galvanic vestibular stimulation. AB - The brain areas activated by bilateral galvanic vestibular stimulation (GVS) were studied using functional magnetic resonance imaging. In six human volunteers, GVS led to activation in the region of the temporoparietal junction, the central sulcus, and the anterior interior intraparietal sulcus, which may correspond to macaque areas PIVC, 3aV, and 2v, respectively. In addition, activation was found in premotor regions of the frontal lobe, presumably analogous to areas 6pa and 8a in the monkey. Since these areas were not detected in previous studies using caloric vestibular stimulation, they could be related to the modulation of otolith afferent activity by GVS. However, the simple paradigm used did not allow separation of the otolithic and semicircular canal effects of GVS. Further studies must be performed to clarify the question of cortical representation of the otolithic information in the human and monkey brain. PMID- 10372082 TI - The interaction of otolith and proprioceptive information in the perception of verticality. The effects of labyrinthine and CNS disease. AB - A review of recent experiments in patients with labyrinthine and neurological disorders assessing the subjective postural vertical (SPV) and the subjective visual vertical (SVV) is presented. The SPV was measured with subjects (Ss) seated in a motorized flight simulator tilting at 1.5 deg/s in roll and pitch; the Ss' task was to indicate when they entered and left self-verticality. The SVV was measured by Ss adjusting a straight line to what they perceived as gravitational upright. Clear dissociations between the SVV and SPV were found, for example, patients with acute unilateral vestibular disorders had marked tilts of the SVV toward the side of the lesion but a "lean" (bias, tilt) of the SPV was never found. Dissociations of the SPV and SVV could also be induced in normal subjects by roll-plane visual motion stimuli: the SVV was tilted in the direction of motion, but the SPV was not. Prolonged lateral body tilt did, however, bias the SVV (the "A" effect) and the SPV, but these effects are likely to be mediated by somatosensory rather than otolithic input. Evidence for the latter came from (i) findings in patients with absent vestibular function, who showed an enhanced "A" effect, and (ii) from a patient with a thalamic infarction, who showed absence of the "A" effect when leaning on the hemihypesthetic side. In separate experiments where normal Ss indicated space-vertical and space-horizontal with saccadic eye movements, we found differences between these percepts, that is, subjective external space lost orthogonality. The findings in these various experiments can be interpreted if we abandon the idea of a single, "internal representation" of verticality. Different sensory modalities convey different and sometimes conflicting messages about verticality. Otolithic and somatosensory signals can have opposite sign effects during verticality estimates while tilted. In man, somatosensory cues have a prominent role in verticality perception. PMID- 10372083 TI - The role of the otoliths in perception of the vertical and in path integration. AB - The role of the otoliths in essential performances of human orientation is analyzed. The following interactions of the otoliths are considered: 1. The otoliths cooperate with graviceptors in the trunk in the perception of body posture. The truncal graviceptors turn out to yield on average 60% of the total gain. 2. The otoliths cooperate with proprioceptors in the head-to-trunk coordinate transformation. However, under static conditions, proprioceptors in the legs, although effective in the control of posture, neither affect the perception of posture nor of the visual vertical. 3. In contrast to the perception of posture, the perception of the visual vertical (SVV) receives the necessary gravity information exclusively from the otoliths. However, their output appears to be affected by a central nervous component that tends to rotate the SVV into the z-axis of head and trunk. A theory of vectorial summation of this component, the "idiotropic vector," with the otolithic vector is able to explain the cause of the A- and E- effects, the increase of the variance of the SVV with the tilt angle, and the asymmetrical effect of rotatory visual flow. 4. Finally, it is shown that the otoliths, by the separation of the effects of tilt from those of translation, play an essential role in navigation by path integration. PMID- 10372084 TI - Replication of passive whole-body linear displacements from inertial cues. Facts and mechanisms. AB - Using path integration, normal subjects should be able to compute the distance of a traveled path even from the sole inertial sensory input. Blindfolded subjects were submitted to a passive linear forward displacement along 2 to 10 m. Their task was to replicate the traveled distance, still blindfolded, by driving the vehicle they were seated upon using a joystick that controlled linear speed. Subjects replicated both the length and the velocity profile of the passive travel, suggesting that a dynamic record of experienced motion is stored in memory. Even when the replication of passive motion dynamics was made impossible, the subjects could still replicate the displacement. The results are explained by a dynamic feedback model that performs a running comparison between the perceived instantaneous displacement of the ongoing motion and the displacement derived from a spatiotemporal record of perceived passive motion. A multimodal acceleration-related sensory input is transformed into a displacement-related perception through double time-integration. PMID- 10372085 TI - Artificial gravity considerations for a mars exploration mission. AB - Artificial gravity (AG), as a means of preventing physiological deconditioning of astronauts during long-duration space flights, presents certain special challenges to the otolith organs and the adaptive capabilities of the CNS. The key issues regarding the choice of AG acceleration, radius, and rotation rate are reviewed from the viewpoints of physiological requirements and human factors disturbances. Head movements and resultant Coriolis forces on the rotating platform may limit the usefulness of economical short centrifuges for other than brief periods of intermittent stimulation. PMID- 10372086 TI - The role of somatosensory input for the perception of verticality. PMID- 10372087 TI - Otolith-vestibular-evoked potentials in humans. Intensity, direction of acceleration (Z+, Z-), and BESA modeling of generators. PMID- 10372088 TI - Measuring the otolith-ocular response by means of unilateral radial acceleration. PMID- 10372089 TI - Saccular dysfunction in Meniere's patients. A vestibular-evoked myogenic potential study. PMID- 10372090 TI - VOR gain modulation in the monkey due to convergence of otolith and semicircular canal afferences during eccentric sinusoidal rotation. PMID- 10372091 TI - An alternative approach to the central processing of canal and otolith signals. PMID- 10372092 TI - Otolith signal processing and motion sickness. PMID- 10372093 TI - Otolith-canal interaction during pitch while rotating. PMID- 10372094 TI - Phase adaptation of the linear vestibulo-ocular reflex. PMID- 10372095 TI - Separation between on- and off-center passive motion in darkness. PMID- 10372096 TI - Development of synaptic innervation in the rodent utricle. PMID- 10372097 TI - Human gaze stabilization for voluntary off-centric head rotations. PMID- 10372098 TI - A simple model of vestibular canal-otolith signal fusion. PMID- 10372099 TI - Centrifugal force affects perception but not nystagmus in passive rotation. PMID- 10372100 TI - Oculomotor, postural, and perceptual asymmetries associated with a common cause. Craniofacial asymmetries and asymmetries in vestibular organ anatomy. PMID- 10372101 TI - Role of otoliths in spatial orientation during passive travel in a curve. PMID- 10372102 TI - Comparison of tilt estimates based on line settings, saccadic pointing, and verbal reports. PMID- 10372103 TI - Vestibular projections in the human cortex. PMID- 10372104 TI - Spatial properties of otolith units recorded in the vestibular nuclei. PMID- 10372105 TI - Introduction to stem cell biology in vitro. Threshold to the future. AB - Transplantable hematopoietic cells with multilineage reconstituting ability can be quantitated in suspensions of human or murine cells using similar assay procedures. The incorporation into these assays of stringently defined functional endpoints ensures a high degree of specificity for the cells detected. Application of these assays to stem cell-containing suspensions after they have been stimulated for several days with defined cytokines in vitro, or by a mixture of defined and/or undefined factors in vivo, has shown that net amplifications in these populations can be obtained under both circumstances. Such studies have allowed cytokine conditions that support stem cell self-renewal divisions to be identified and have also provided evidence that stem cell regeneration can be manipulated both in vitro and in vivo by altering the molecular milieu of the responding cells. These observations pave the way to future delineation of mechanisms that control the normal behavior, pathology and future clinical exploitation of hematopoietic stem cell populations. PMID- 10372106 TI - Hematopoietic commitment during embryogenesis. AB - Hematopoiesis develops initially as discrete blood islands in the extraembryonic yolk sac of the embryo. These blood islands consist of clusters of primitive erythrocytes surrounded by developing angioblasts that ultimately form the yolk sac vasculature. The close developmental association of these early hematopoietic and endothelial cells has led to the hypothesis that they develop from a common precursor, a cell known as the hemangioblast. Using a developmental model system based on the in vitro differentiation capacity of embyronic stem (ES) cells, we have identified a precursor with the capacity to generate endothelial as well as primitive and definitive hematopoietic progeny. The developmental potential of this precursor population suggests that it represents the in vitro equivalent of the hemangioblast. PMID- 10372107 TI - Humoral regulation of hematopoietic stem cells. AB - During the last two decades, studies using primarily cell culture methods disclosed that a number of hematopoietic cytokines possess stimulatory effects on primitive hematopoietic progenitors. More recently, investigators in a number of laboratories, including ours, used murine transplantation models to characterize the cytokines regulating the hematopoietic stem cells. The results are in general agreement with the cytokine interactions defined in culture. The positive cytokines may be separated into two groups: one consisting of steel factor and flt3/flt2 ligand and the other consisting of interleukin (IL)-6, IL-11, IL-12, leukemia inhibitory factor, granulocyte colony-stimulating factor (G-CSF) and thrombopoietin. Interactions of two cytokines belonging to different groups appear necessary to positively regulate the kinetics of stem cells. Surprisingly, IL-3 and IL-1 proved to have profound negative effects on hematopoietic stem cells. Studies of human hematopoietic stem cells are now necessary. PMID- 10372108 TI - Expression of novel surface antigens on early hematopoietic cells. AB - The purpose of this report is to demonstrate the expression of very recently identified surface antigens on CD34+ and AC133+ bone marrow (BM) cells. Coexpression analysis of AC133 and defined antigens on CD34+ BM cells revealed that the majority of the CD164+, CD135+, CD117+, CD38low, CD33+, and CD71low cells resides in the AC133+ population. In contrast, most of the CD10+ and CD19+ B cell progenitors and a fraction of the CD71high population are AC133-, indicating that CD34+AC133+ cells are enriched in primitive and myeloid progenitor cells, whereas CD34+AC133- cells mainly consist of B cell and late erythroid progenitors. This corresponds to the highly reduced percentage of CD10+ B cells and the absence of CD71high erythroid progenitors on AC133+ selected BM cells. A portion of 0.2-0.7% of the AC133+ selected cells do not coexpress CD34. These cells are very small and define a uniform CD71-, CD117-, CD10-, CD38low, CD135+, HLA-DRhigh, CD45+ population with unknown delineation. Four color analysis on CD34+CD38- BM cells revealed that virtually all of these primitive cells express AC133. Using an improved liposome-enhanced labeling technique for the staining of weakly expressed antigens, subsets of this population could be identified which express the angiopoietin receptors TIE (67.6%) and TEK (36.8%), the vascular endothelial growth factor receptors FLT1 (7%), FLT4 (3.2%), and KDR (10.4%), or the receptor tyrosine kinases HER-2 (15.4%) and FLT3 (CD135; 77.6%). Our results suggest that the CD34+CD38- population is heterogeneous with respect to the expression of the analyzed receptor tyrosine kinases. PMID- 10372109 TI - Lymphohematopoietic stem cell engraftment. AB - Traditional dogma has stated that space needs to be opened by cytoxic myeloablative therapy in order for marrow stem cells to engraft. Recent work in murine transplant models, however, indicates that engraftment is determined by the ratio of donor to host stem cells, i.e., stem cell competition. One hundred centigray whole body irradiation is stem cell toxic and nonmyelotoxic, thus allowing for higher donor chimerism in a murine syngeneic transplant setting. This nontoxic stem cell transplantation can be applied to allogeneic transplant with the addition of a tolerizing step; in this case presensitization with donor spleen cells and administration of CD40 ligand antibody to block costimulation. The stem cells that engraft in the nonmyeloablated are in G0, but are rapidly induced (by 12 hours) to enter the S phase after in vivo engraftment. Exposure of murine marrow to cytokines (IL-3, IL-6, IL-11 and steel factor) expands progenitor clones, induces stem cells into cell cycle, and causes a fluctuating engraftment phenotype tied to phase of cell cycle. These data indicate that the concepts of stem cell competition and fluctuation of stem cell phenotype with cell cycle transit should underlie any new stem cell engraftment strategy. PMID- 10372110 TI - Homing of long-term and short-term engrafting cells in vivo. AB - Long-term repopulating hematopoietic stem cells can be separated from cells which provided radioprotection (short-term repopulating cells) on the basis of size. This might be a result of the quiescent nature of long-term repopulating cells. To define the activity of these populations we utilized a dye, PKH26, which incorporates into the membrane of cells and is equally distributed to daughter cells when they divide. We were able to retrieve PKH26(+)-labeled cells posttransplant in the hematopoietic tissues of the recipients. We could also assess their cell cycle status and their ability, short- and long-term, to reconstitute secondary lethally irradiated hosts in limiting dilution. The results suggest that long-term repopulating cells remain quiescent in the bone marrow shortly after engraftment, whereas cells which radioprotect are more rapidly dividing. We could not detect labeled cells in the peripheral blood posttransplant, and even though cells homed to both the spleen and bone marrow the cells in the bone marrow were significantly more competent at reconstituting lethally irradiated secondary hosts. PMID- 10372111 TI - Further characterization of CD34-low/negative mouse hematopoietic stem cells. AB - We have previously reported that in adult mouse bone marrow, CD34low/- c-kit+ Sca 1+ lineage markers negative (Lin-) (CD34-KSL) cells represent hematopoietic stem cells with long-term marrow repopulating ability whereas CD34+ c-kit+ Sca-1+ Lin- (CD34+KSL) cells are progenitors with short-term reconstitution capacity. To further characterize cells in those two populations, relative expression of various genes were examined by reverse transcriptase polymerase chain reaction (RT-PCR). In CD34-KSL cells, none of the genes studied was found to be expressed with the exception of GATA-2, IL-1R alpha, IL-2R gamma, AIC-2B, c-kit, EPO-R, and c-mpl. In contrast, expression of GATA-1 and all cytokine receptor genes examined except IL-2R beta, IL-7R alpha and IL-9R alpha were found in CD34+KSL. The difference between these two populations was also shown in single cell culture analysis of these cells. When cells were clone-sorted and cultured in the presence of SCF, IL-3 and EPO, CD34-KSL cells required much more time to undergo the first cell division than CD34+KSL cells. Dormancy and random fashion of cell division by CD34-KSL cells were also evident by the analysis of the second cell division, which was found to be delayed and unsynchronous compared with CD34+KSL cells. Clonal culture analysis showed that CD34-KSL cells were more potent in proliferation and multilineage differentiation capacities than CD34+KSL cells. In a paired-daughter cell experiment, 75% of CD34-KSL and 50% of CD34+KSL paired daughter-derived colonies were nonidentical with wide variety of lineage combinations. Taken together, these data support our previous notion that CD34 KSL cells are at higher rank in hematopoietic hierarchy than CD34+KSL cells. In addition, our results using highly enriched stem cell population directly obtained from mouse bone marrow support the proposed stochastic nature of lineage commitment. PMID- 10372112 TI - Biology of IL-8-induced stem cell mobilization. AB - The CXC chemokine interleukin-8 (IL-8) has profound hematopoietic activities following systemic administration. It induces the rapid mobilization of cells with lymphomyeloid repopulating ability in mice and of hematopoietic progenitor cells in monkeys. In this paper, evidence is presented that stem cell mobilization in mice requires the functional expression on the beta 2-integrin leukocyte function-associated antigen-1 (LFA-1). In monkeys, systemic injection of IL-8 is followed by a significant increase in the circulating levels of the matrix metallo proteinase gelatinase-B (MMP-9). Based on these findings, the hypothesis is discussed that mature neutrophils serve as intermediate cells in IL 8-induced stem cell mobilization by the release of proteinases. PMID- 10372113 TI - (R)evolutionary considerations in hematopoietic development. AB - Evolutionary aspects of three characteristics of the mammalian hematopoietic system are considered in the context of both established and recent data. First, the lineage relationships among early members of the hematopoietic hierarchy are reconsidered in a tripartite model proposing lineage segregation based on vascular function, innate immunity, and acquired immunity on an evolutionary time scale. Second, the observation of two stem cell populations that differ in cell cycle status is considered as an evolved mechanism to enhance survival of the species in response to exposure to environmental toxins. Finally, the mobilization of hematopoietic stem cells into the peripheral circulation is proposed to be a mechanism for rapid dissemination of myeloid function during acute bacterial infections. These revolutionary hypotheses challenge some conventional concepts of stem cell biology, and provide an evolutionary context for considering mammalian hematopoiesis. PMID- 10372114 TI - Stem cell gene therapy for the beta-chain hemoglobinopathies. Problems and progress. AB - Virus vectors hold great promise for the stem cell gene therapy of beta-chain hemoglobinopathies. However, conventional vectors suffer from low gene transfer rates, low expression levels, and inconsistent or short-lived expression in vivo. In this review we summarize the current status of vector systems for the transduction of hematopoietic stem cells, including the development of novel vector systems and methods for selection of transduced stem cells in vivo. We also summarize efforts to achieve therapeutic expression levels of transferred globin genes with retrovirus vectors, including the manipulation of transcription cassettes, the use of globin gene enhancers, and advances in the use of chromatin insulators for improving the frequency of gene expression following hematopoietic stem cell transduction. PMID- 10372115 TI - Retroviral-fibronectin interactions in transduction of mammalian cells. AB - Hematopoiesis occurs in a complex environment in the medullary cavity in close proximity to stromal cells, fibroblasts, endothelial cells and matrix molecules. Hematopoietic cell interactions in this environment appear to involve both integrin and proteoglycan-mediated cell-cell and cell-matrix interactions. Genetic transduction of hematopoietic stem cells via retroviral vectors has been hampered by low efficiency of gene transfer. Recently, hematopoietic stem cell adhesion to the extracellular matrix molecule fibronectin has been shown to increase transduction of these target cells using retrovirus vectors. The mechanism of increased transduction appears to involve colocalization of virus particles and target cells. These data are reviewed in this paper. PMID- 10372116 TI - Amphotropic retrovirus transduction of hematopoietic stem cells. AB - Mice treated with cytokines for 5 days have large numbers of hematopoietic stem cells (HSCs) in their peripheral blood and bone marrow at 1 and 14 days after the last injection. We fractionated the HSCs from the bone marrow of these mice using elutriation at flow rates of 25, 30 and 35 ml/min. The subpopulations of HSCs from cytokine-treated mice show a 3- to 8-fold higher level of mRNA encoding the amphotropic retrovirus receptor (amphoR) compared with the corresponding HSC subpopulation from untreated mouse bone marrow. In an earlier study with mouse HSCs we showed a direct correlation between high levels of amphoR mRNA and efficient retrovirus transduction. We have now utilized our gene transfer protocol to assay amphotropic retrovirus transduction efficiency using HSCs from the bone marrow of mice treated with granulocyte-colony stimulating factor/stem cell factor (G-CSF/SCF). To extend these findings to a more clinically relevant protocol we analyzed the amphoR mRNA levels in HSCs from human cord blood and adult bone marrow. The amphoR mRNA level in HSCs from human bone marrow and fresh cord blood was detectable at an extremely low level compared with the HSC population in cryopreserved cord blood samples. The 12- to 22-fold increase in amphoR mRNA in HSCs from cryopreserved cord blood renders these HSCs likely candidates for high efficiency, gene transfer. PMID- 10372117 TI - Effects of retroviral-mediated MDR1 expression on hematopoietic stem cell self renewal and differentiation in culture. AB - Ex vivo expansion of hematopoietic stem cells would be useful for bone marrow transplantation and gene therapy applications. Toward this goal, we have investigated whether retrovirally-transduced murine stem cells could be expanded in culture with hematopoietic cytokines. Bone marrow cells were transduced with retroviral vectors expressing either the human multidrug resistance 1 gene (HaMDR1), a variant of human dihydrofolate reductase (HaDHFR), or both MDR1 and DHFR in an internal ribosomal entry site (IRES)-containing bicistronic vector (HaMID). Cells were then expanded for 15 days in cultures stimulated with interleukin (IL)-3, IL-6, and stem cell factor. When very low marrow volumes were injected into lethally irradiated recipient mice, long-term reconstitution with 100% donor cells was seen in all mice injected with HaMDR1- or HaMID-transduced cells. By contrast, engraftment with HaDHFR- or mock-transduced cells ranged from partial to undetectable despite injection of significantly larger marrow volumes. In addition, mice transplanted with expanded HaMDR1- or HaMID-transduced stem cells developed a myeloproliferative disorder that was characterized by an increase in abnormal peripheral blood leukocytes. These results show that MDR1 transduced stem cells can be expanded in vitro with hematopoietic cytokines, but indicate that an increased stem cell division frequency can lead to stem cell damage. PMID- 10372118 TI - Effects of CC, CXC, C, and CX3C chemokines on proliferation of myeloid progenitor cells, and insights into SDF-1-induced chemotaxis of progenitors. AB - Chemokines have been implicated in the regulation of stem/progenitor cell proliferation and movement. The purpose of the present study was to assess a number of new chemokines for suppressive activity and to delve further into SDF-1 mediated chemotaxis of progenitor cells. This report extends the list of chemokines that have suppressive activity against immature subsets of myeloid progenitors stimulated to proliferate by multiple growth factors to include: MCP 4/CK beta-10, MIP-4/CK beta-7, I-309, TECK, GCP-2, MIG and lymphotactin. The suppressive activity of a number of other chemokines was confirmed. Additionally, pretreatment of the active chemokines with an acetylnitrile solution enhanced specific activity of a number of these chemokines. The new chemokines found to be lacking suppressive activity include: MCP-2, MCP-3, eotaxin-1, MCIF/HCC-1/CK beta 1, TARC, MDC, MPIF-2/eotaxin-2/CK beta-6, SDF-1 and fractalkine/neurotactin. Overall, 19 chemokines, crossing the CC, CXC, and C subgroups, have now been found to be myelosuppressive, and 14 chemokines crossing the CC, CXC and CX3C subgroups have been found to lack myelosuppressive activity under the culture conditions of our assays. Because of the redundancy in chemokine/chemokine receptor interactions, it is not yet clear through which chemokine receptors many of these chemokines signal to elicit suppressive activities. It was also found that SDF-1-induced chemotaxis of progenitors can occur in the presence of fibronectin (FN) and extracellular matrix components and that FN effects involve activation of beta 1-, and possibly alpha 4-, integrins. PMID- 10372119 TI - Dissecting the marrow microenvironment. AB - Cloned human stromal cell lines representing functionally distinct cellular components of the marrow microenvironment were generated to serve as tools for identifying gene products that regulate hematopoiesis. Oligonucleotide arrays, or "gene chips" were used to provide a comprehensive comparison of gene expression among the cell lines. One line, designated HS-5, was found to secrete large amounts of cytokines, and conditioned media from this line was found to support the ex vivo expansion of both immature and mature progenitors. In contrast, a second line, designated HS-27a, does not secrete known cytokines but does support cobblestone area formation by CD34+/38lo cells. HS-27a, but not HS-5, was also found to express hJagged1, a ligand for Notch1, which may function to influence cell fate decisions of hematopoietic precursors. Both cell lines are currently being used to identify other gene products that regulate hematopoiesis and to generate reagents that will allow more formal evaluation of the putative role of hJagged1 in hematopoietic cell fate decisions. PMID- 10372120 TI - Stromal inhibition of myeloid differentiation. A possible role for hJagged1. PMID- 10372121 TI - Regulation of transendothelial migration of hematopoietic progenitor cells. AB - Transendothelial migration of hematopoietic progenitor cells occurs in the bone marrow during mobilization and homing, and may therefore play a key role in the trafficking of hematopoietic stem cells. We hypothesize that adhesion molecules, chemokines, and paracrine cytokines are involved in this multifactorial process. As suggested in several studies, downregulation of adhesion molecules (e.g., integrins) may contribute to mobilization of progenitors due to a decreased avidity to bone marrow stromal and endothelial cells, which express the corresponding ligands. Using an in vitro model of transendothelial migration, we have shown that only a small number of more mature, committed progenitors migrates spontaneously under the control of adhesion molecules of the beta-2 integrin family and their corresponding endothelial/stromal ligands. However, transendothelial migration of progenitors in vitro is substantially enhanced by the chemokine stromal cell-derived factor-1 (SDF-1), which is constitutively produced by bone marrow stromal cells. More primitive progenitors also respond to this chemokine. In addition, the ligand for SDF-1, the chemokine receptor CXCR-4, is expressed in greater levels on bone marrow CD34+ cells as compared to mobilized progenitors, suggesting that downregulation of chemokine receptors occurs during progenitor mobilization. Indeed, bone marrow CD34+ cells migrate more avidly in response to SDF-1 than mobilized progenitors. Paracrine cytokines may also play a role in hematopoietic stem cell trafficking, since growth factor stimulated hematopoietic cells produce cytokines that act on endothelial cells (e.g., vascular endothelial growth factor, VEGF), modifying their proliferation, motility, permeability, and fenestration. We conclude that transendothelial migration of hematopoietic progenitor cells is regulated by adhesion molecules, paracrine cytokines, and chemokines. Cytotoxic therapy as well as exogenously administered hematopoietic growth factors may affect adhesion molecule expression, the local cytokine and chemokine milieu, and chemokine receptor expression, which indirectly results in mobilization of hematopoietic stem cells. PMID- 10372122 TI - Hematopoietic stem/progenitor cell mobilization. A continuing quest for etiologic mechanisms. AB - The physiologic egress of mature hemopoietic cells and of hemopoietic stem/progenitor cells from bone marrow to the circulation are poorly understood processes. Likewise, the mechanism of their enforced emigration or mobilization through the use of several agents has not been unraveled. Although mobilization is suspected to be a multi-step process, involving sequential and/or overlapping changes in adhesion and migratory capacity, a model of molecular hierarchy, like the one governing the extravasation of mature leukocytes to tissues of inflammation, has not been worked out. Understanding the in vivo mechanism of mobilization has been a challenge. Signals emanating from both stromal cells and from hemopoietic cells are likely involved. However, dissecting out their roles, specificity, and interactions has been difficult. Nevertheless insightful information is rapidly emerging, especially with the current availability of many mouse models bearing targeted disruptions of cytoadhesion or signaling molecules. PMID- 10372124 TI - Absence of CD34 on some human SCID-repopulating cells. AB - The availability of in vivo repopulation assays has greatly aided the study of human hematopoietic stem cells. Here, I shall review recent data that has identified a novel class of human repopulating cells that do not express classical stem cell markers including CD34 but still retain the ability to repopulate nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice. PMID- 10372123 TI - Characterization of purified and ex vivo manipulated human hematopoietic progenitor and stem cells in xenograft recipients. AB - Research on the biology, regulation, and transplantation of human hematopoietic stem cells requires test systems for the detection, monitoring, and quantitation of these cells. Xenografted animal models provide suitable stem cell assays, since they allow long-term engraftment, multilineage differentiation, and serial transfer of human hematopoietic cells. Recent techniques for the separation of hematopoietic cells have provided highly purified cellular subsets selected on the basis of the surface marker phenotype. The stem cell content of these subsets, however, is still unclear. Also, innovative approaches for the induction of hematopoietic cell proliferation and differentiation have generated ex vivo manipulated cells whose biological properties and functions still remain to be assessed. This paper reports on the biological characterization of these cell populations by the use of xenograft models. PMID- 10372125 TI - Engraftment and multilineage expression of human bone marrow CD34- cells in vivo. AB - The fetal sheep competitive engraftment model of human hematopoietic stem cells (HSC) was used to evaluate the in vivo engraftment potential of human bone marrow CD34- Lin- cells. Transplantation of CD34- Lin- cells into primary hosts resulted in the long-term (> 1 year) engraftment and multilineage donor cell/progenitor expression with production of significant numbers of CD34+ cells. Secondary transplantation and limiting dilution studies confirmed the presence in human CD34- fraction of HSC with in vivo long-term engraftment and multilineage differentiation potentials. PMID- 10372126 TI - Ex vivo expansion and genetic marking of primitive human and baboon hematopoietic cells. AB - The achievement of positive outcomes in many clinical protocols involving hematopoietic stem cells (HSCs) has been handicapped by the limited numbers of marrow repopulating cells available to actually bring about therapy. This insufficiency has been especially problematic in stem cell transplantation and gene therapy. A number of studies have been initiated to attempt expansion of HSCs, mainly by manipulation of key cytokines in cell suspension cultures. Unfortunately, these expansion methods usually lead to altered properties in the amplified cells, mainly by reducing their self-renewal and multi-lineage differentiative potentials. Here we discuss our ongoing work, utilizing a unique endothelial cell line that supports primitive hematopoiesis, to attempt to generate expansion of primate HSCs that retain their elementary properties. Genetic marking of early hematopoietic cells to facilitate tracking will be mentioned as will the development and employment of assay systems designed to evaluate the long-term functional attributes of the expanded cells. PMID- 10372127 TI - Isolation of stem cell-specific cDNAs from hematopoietic stem cell populations. AB - We have begun to isolate gene sequences that are specifically expressed in hematopoietic stem cells (HSCs). There are at least three fundamental requirements for the isolation of HSC-specific transcripts. First, highly enriched populations of HSCs, and an HSC-depleted cell population for comparison must be isolated. Secondly, the gene isolation procedures must be adapted to accommodate the small amounts of RNA obtained from purified HSCs. Finally, a defined screening strategy must be developed to focus on sequences to be examined in more detail. In this report, we describe the characterization of populations of HSCs that are highly enriched (Lin- c-kitHI) or depleted (Lin- c-kitNEG) of HSCs. We compared two methods for gene isolation, differential display polymerase chain reaction (DD-PCR) and subtractive hybridization (SH), and found that the latter was more powerful and efficient in our hands. Lastly we describe the strategy that we have developed to screen clones for further study. PMID- 10372128 TI - Embryonic beginnings of definitive hematopoietic stem cells. AB - The ability of the many cell types within the adult blood system to be constantly replenished and renewed from hematopoietic stem cells is an interesting problem in development and differentiation and has led to questions concerning how, when and where these stem cells for the adult hematopoietic system are generated within the embryo. During embryonic development many mature hematopoietic cells appear before adult-type hematopoietic stem cells thus the notion of a conventional hematopoietic hierarchy is challenged. Experiments probing the development of hematopoietic stem cells in the mouse embryo strongly suggest that at least two independent hematopoietic sites generate blood cells during development; the yolk sac, which produces the transient embryonic hematopoietic system, and the AGM (aorta-gonad-mesonephros) region, which initiates the long lived adult hematopoietic system. PMID- 10372129 TI - Asymmetric cell divisions in hematopoietic stem cells. AB - In order to study cell kinetics involved in long-term hematopoiesis, we studied single sorted candidate hematopoietic stem cells (HSC) from fetal liver cultured in the presence of a mixture of stimulatory cytokines. After 8-10 days in culture, the number of cells varied from less than a hundred to more than ten thousand cells. Single cells in slowly growing colonies were recloned upon reaching a 100-200-cell stage. Strikingly, the number of cells in subclones varied widely again. These results are indicative of asymmetric divisions in primitive hematopoietic cells in which the proliferative potential and cell cycle properties are unevenly distributed among daughter cells. The continuous generation of heterogeneity in cell cycle properties among the clonal progeny of HSC appears a relevant mechanism to maintain long-term maintenance of hematopoiesis in vitro and in vivo. PMID- 10372130 TI - Searching for stem cell regulatory molecules. Some general thoughts and possible approaches. AB - Hematopoietic development in the mammal can be represented as a numerically expanding hierarchy of cell populations that are progressively restricted in their self-renewal and differentiation abilities. Classical functional studies have now been extended to provide exact physical descriptions of various stages in the hematopoietic hierarchy. In particular, much information is available that defines the properties of the most primitive stem cell compartment. In addition, a number of in vitro culture systems suggest the possibility of maintaining and expanding these cells in a defined context. In all developmental systems, unique profiles of expressed genes define distinct differentiation stages. Within these profiles are gene products that play crucial roles in the regulation of cell-fate decisions. Recent progress in hematopoietic biology provides the framework within which to define molecular phenotypes for hematopoietic stem cells and their immediate clonal progeny. Identifying novel gene products expressed predominantly in uncommitted stem cells together with functional loss and gain-of-function approaches should begin to unravel the molecular mechanisms that govern biological phenomena such as self-renewal, commitment, and proliferation in the hematopoietic system. PMID- 10372131 TI - Stem cells, pre-progenitor cells and lineage-committed cells: are our dogmas correct? AB - Recent developments warrant careful reexamination of several of the central dogmas of hematopoiesis. The bioassays previously used may have predetermined which subsets of hematopoietic stem cells are regarded as having long-term repopulating activity and thus have produced misleading data. Lineage commitment in multipotential cells has been regarded as an immutable stochastic process but may be a process that can be modified by extrinsic signaling. Finally, loss of self-renewal activity has been regarded as progressive and irreversible but this response to signaling can be blocked by cytokine-inducible modulating proteins. PMID- 10372132 TI - Signaling domains of the beta c chain of the GM-CSF/IL-3/IL-5 receptor. AB - The granulocyte/macrophage colony-stimulating factor (GM-CSF)/interleukin-3 (IL 3)/IL-5 receptors are a family of heterodimeric transmembrane proteins expressed by myeloid lineage cells. Each receptor has a unique ligand-binding alpha chain and they share a common beta chain (beta c chain). Binding of GM-CSF activates at least one receptor-associated tyrosine kinase, JAK2, and rapidly induces tyrosine phosphorylation of the GMR beta c chain (GMR beta), but not the GMR alpha chain (GMR alpha). Mutation of each of the 8 tyrosine residues in the cytoplasmic domain of the human GMR beta to phenylalanine (GMR beta-F8) reduced tyrosine phosphorylation of GMR beta, SHP2 and SHC, but not JAK2 or STAT5. Interestingly, GMR beta-F8 was still capable of inducing at least short-term proliferation and enhancing viability. The role of each individual tyrosine residue was explored by replacing each mutated phenylalanine with the wild-type tyrosine residue. Tyrosine 577 was found to be sufficient to regenerate GM-CSF-dependent phosphorylation of SHC, and any of Y577, Y612, or Y695 were sufficient to regenerate GM-CSF-inducible phosphorylation of SHP2. Next, a series of four internal deletion mutants were generated, which deleted small sections from aa 518 to 626. One of these, deleting residues 566-589 was profoundly defective in signaling and supporting viability, and may identify an important viability signaling domain for this receptor family. Overall, these results indicate that GMR beta tyrosine residues are not necessary for activation of the JAK/STAT pathway, or for proliferation, viability, or adhesion signaling in Ba/F3 cells, although tyrosine residues significantly affect the magnitude of the response. However, internal deletion mutant studies identify critical domains for viability and proliferation. PMID- 10372133 TI - Thrombopoietin and hematopoietic stem cell development. AB - Thrombopoietin, the long sought primary regulator of thrombopoiesis, was cloned four years ago. In addition to its fulfilling most, if not all, of the expected biological activities relating to megakaryocyte and platelet development, the availability of the recombinant hormone and reagents to characterize its receptor have allowed detailed investigation of additional biological activities. In cultures of purified populations of candidate stem cells, thrombopoietin supports the survival, and augments the proliferation of hematopoietic stem cells when present together with interleukin-3 or steel factor. The progeny of such cultures are not skewed in their developmental potential; colony-forming cells of all lineages arise from thrombopoietin-stimulated stem cells. Evidence for an important effect of thrombopoietin on stem cell physiology in vivo are equally compelling. Genetic elimination of thrombopoietin or its receptor leads to a profound reduction not only of megakaryocytes and platelets, but also of committed myeloid progenitors of all types, primitive progenitors and hematopoietic stem cells. When administered to animals, thrombopoietin profoundly stimulates thrombopoiesis and enhances the number of hematopoietic progenitor cells of all lineages, and when used in most animal models of myelosuppressive therapy, accelerates the recovery of platelet, erythrocyte and leukocyte production. Thus, thrombopoietin appears to be more than a lineage-restricted growth factor. PMID- 10372134 TI - G-CSF receptor mutations in patients with severe congenital neutropenia do not abrogate Jak2 activation and stat1/stat3 translocation. AB - Severe congenital neutropenia (SCN) is an inherited disorder of myelopoiesis, characterized by a maturation arrest at the stage of promyelocytes and myelocytes in bone marrow, and absence or low levels of mature neutrophil granulocytes in peripheral blood. Recently, studies of patients with SCN who subsequently developed acute myeloid leukemia (AML) revealed nonsense mutations in the cytoplasmic domain of the granulocyte colony-stimulating factor (G-CSF) receptor messenger RNA. We focused our interest on the G-CSF-mediated signaling cascade to examine the consequences of the observed point mutations for the nuclear translocation of the transcription factors Stat1 and Stat3. Expression vectors encoding for truncated G-CSF receptors were transfected in the murine fibroblast cell line C243 expressing a fusion protein consisting of the transcription factor Stat1 and Stat3, respectively, and the green fluorescent protein (GFP). Nuclear translocation of the GFP fusion proteins was examined after G-CSF stimulation of the transfected cells. PMID- 10372135 TI - The Placental/Umbilical Cord Blood Program of the New York Blood Center. A progress report. AB - The transplantation of placental/umbilical cord blood (P/CB) has been used successfully to reconstitute bone marrow function in both related and unrelated recipients. We report here the experience of the New York Blood Center P/CB Program. Since its inception in 1992, over 400 unrelated transplants were supported between July 1993 and September 1997. Overall, event-free survival for all diagnoses and ages approached 0.45. Success and rapidity of engraftment correlated most strongly with the degree of human leukocyte antigen (HLA) disparity and cell dose/kg body weight recipient. Acute graft-versus-host disease (GVHD) was common in all patients but, surprisingly, did not differ between those patients who received grafts having one or more antigen mismatches. Chronic GVHD was uncommon and only rarely contributed to death. These results demonstrate the feasibility of large-scale P/CB banking for the provision of cryopreserved stem cell preparations for unrelated transplants. The degree of the program's success argues strongly for additional P/CB banks in order to increase the likelihood of finding a suitable stem cell preparation for patients for whom related matched donors do not exist. PMID- 10372136 TI - The role of megadose CD34+ progenitor cells in the treatment of leukemia patients without a matched donor and in tolerance induction for organ transplantation. AB - Throughout the 1980s, transplantation of unmodified (T cell-replete) bone marrow from full haplotype incompatible family donors was associated with an unsuccessful outcome because of graft failure and severe graft-versus-host disease (GVHD), at times affecting up to 90% of recipients. Although extensive T cell depletion of donor bone marrow was successful in preventing GVHD in children with severe combined immunodeficiency disease (SCID), results were disappointing in leukemic patients because the benefit of preventing GVHD was offset by graft failure. Resistance to engraftment appears to be mediated by host-derived cytotoxic T-lymphocyte precursors that survive supralethal conditioning. In the present paper, we review data that show that these genetic histocompatibility barriers can be overcome in stringent mouse models, employing lethally as well as sublethally irradiated recipients, by two major approaches that are synergistic to each other: escalation of hematopoietic progenitor cell dose and the use of nonalloreactive T cells. The former approach is already being successfully implemented in the treatment of leukemic patients. PMID- 10372137 TI - Transplantation of megadoses of purified haploidentical stem cells. AB - Peripheral mobilized parental CD34+ progenitors were isolated and used for the hematopoietic reconstitution after a myeloablative therapy in 23 pediatric patients with various diseases. Fourteen donors were human leukocyte antigen (HLA) three-loci mismatches, 6 donors were two-loci and 3 donors were one-locus mismatches. For depletion of T-lymphocytes, a positive selection of the mobilized peripheral CD34+ progenitors using the method of magnetic-activated cell sorting (MACS) was used. The purity of the CD34+ cells after MACS-sorting was 98-99%, the average number of transplanted CD34+ cells was 14.2 x 10(6)/kg (range 5.4-3.9 x 10(6)/kg) and the average number of infused T-lymphocytes was 1.4 x 10(4)/kg. Due to this low T cell number, only a short-term or no prophylaxis of graft-versus host disease (GVHD) was necessary and no GVHD was seen. A significant GVHD was only seen in patients after add-back of donor T-lymphocytes, which was performed in some patients for prevention of relapse or in patients who showed a transient mixed chimerism. Since the B lymphocyte contamination of the isolated CD34+ cells was low in the range of 0.2%, no Epstein-Barr virus (EBV)-associated lymphoproliferative syndrome was observed. A primary engraftment was seen in 18 patients. Nonengraftment and rejection occurred in three and two patients, respectively. In four of these 5 patients, a second transplant using purified CD34+ cells from the same donor after an immunological reconditioning regimen resulted in a complete and sustained hematopoietic reconstitution. The speed of the immunological recovery was dependent on the number of transplanted CD34+ cells and was more rapid if this number was > 20 x 10(6)/kg. Eleven of the 23 patients are alive and disease free with a median follow-up of 12 months (range 2 30). The main cause of death was relapse (7 patients), and only one fatal infection was seen. Our data suggest that the transplantation of megadoses of haploidentical CD34+ cells is a realistic therapeutic option for patients who otherwise have no suitable donor, and an alternative to the use of unrelated cord blood. PMID- 10372138 TI - Approaches to dendritic cell-based immunotherapy after peripheral blood stem cell transplantation. AB - High-dose chemotherapy with peripheral blood progenitor cell transplantation (PBPCT) is a potentially curative treatment option for patients with both hematological malignancies and solid tumors, including breast cancer. However, based on a number of clinical studies, there is strong evidence that minimal residual disease (MRD) persists after high-dose chemotherapy in a number of patients, which eventually results in disease recurrence. Therefore, several approaches to the treatment of MRD are currently being evaluated, including treatment with dendritic cell (DC)-based cancer vaccines. DCs, which play a crucial role with regard to the initiation of T-lymphocyte responses, can be generated ex vivo either from CD34+ hematopoietic progenitor cells or from blood monocytes. They can be pulsed in vitro with tumor-derived peptides or proteins, and then used as a professional antigen-presenting cell (APC) vaccine for the induction of antigen-specific T-lymphocytes in vivo. This paper summarizes our preclinical studies on the induction of primary HER-2/neu specific cytotoxic T lymphocyte (CTL) responses using peptide-pulsed DC. As HER-2/neu is overexpressed on 30-40% of breast and ovarian cancer cells, this novel vaccination approach might be particularly applicable to advanced breast or ovarian cancer patients after high-dose chemotherapy and autologous PBPCT. PMID- 10372139 TI - Mixed hematopoietic chimerism after marrow allografts. Transplantation in the ambulatory care setting. AB - This paper describes the development of nonmyeloablative marrow transplant programs that have little toxicity in a canine model and their translation to patients with malignant and nonmalignant hematological diseases. PMID- 10372141 TI - Canine lymphocyte expression of retrovirally transferred human common gamma chain. PMID- 10372140 TI - Clinical applications of mixed chimerism. AB - Bone marrow transplantation (BMT) is currently a procedure that is associated with high morbidity and mortality. Thus, the clinical application of this technique is limited to the treatment of life-threatening hematopoietic malignancies. The morbidity and mortality of BMT is mainly related to graft versus-host disease (GVHD), failure of engraftment, and toxicity related to fully myeloablative conditioning. GVHD can be prevented by T-cell depletion. However, T cell depletion increases the risk of failure of engraftment. With the identification of a facilitating cell population that enables engraftment of hematopoietic stem cells across major histocompatibility barriers, the dichotomy between GVHD and failure of engraftment has been resolved. If one could overcome the toxicity of conditioning with the development of partially ablative conditioning strategies, BMT could be used for the treatment of a variety of nonmalignant diseases, as well as in the induction of donor-specific transplantation tolerance. This review outlines the development and advantages of partially ablative conditioning strategies and illustrates possible applications of the technique. Forty years ago E.D. Thomas discussed the potential of BMT for treating immunodeficiencies and for the induction of transplantation tolerance. BMT can be viewed as a natural form of gene therapy to replace a defective cell or enzyme with a functional and normally regulated one. PMID- 10372142 TI - Effects of several cytokine combinations on retrovirus-mediated human MDR1 gene transfer into bone marrow hematopoietic cells. PMID- 10372143 TI - Thrombopoietin and interleukin-3 are chemotactic and chemokinetic chemoattractants for a factor-dependent hematopoietic progenitor cell line. PMID- 10372144 TI - A human factor-dependent cell line simulates cobblestone formation under human bone marrow stromal cells in vitro. PMID- 10372145 TI - Monitoring temperature-induced changes in tissue during hyperthermia by impedance methods. AB - The electrical conduction in living tissue depends on temperature in two ways: (1) the temperature coefficients of conductivity of the intra- and extracellular electrolytes and (2) temperature-induced fluid volume shifts in the tissue. Measurements in rat skeletal muscle and tumors (DS sarcoma) during hyperthermic treatment reveal that the contribution of fluid volume shifts to changes in conductance is of the same order of magnitude as the change in fluid conductivity. In skeletal muscles, blood volume changes are caused by the temperature-dependent regulation of the vessel diameter (vasodilatation). In tumors, fluid content changes irregularly. These effects render temperature measurements by impedance methods, for example, electrical impedance tomography (EIT), questionable. However, monitoring fluid volume changes in tissue and the state of cell membranes is an interesting application of impedance (or admittance) spectroscopy and tomography as well. PMID- 10372146 TI - Application of linear circuit models to impedance spectra in irradiated muscle. AB - We have applied a number of modeling schemes to previously reported in vivo electrical impedance measurements on irradiated and normal muscle in the hind legs of rats. Specifically, seven-parameter parallel pathways and embedded membrane circuit models have been fit to group averages of impedance spectra measured at different doses and time points. Correlations between histologically scored tissue sections and model parameters have also been determined. The results show that both models produce good fits to the experimental observations, especially in the case of the irradiated tissues. The correlations between histology scores and circuit parameters were, however, higher with the embedded model. Trends in the spectra and the model parameters were found to agree with the expected changes in tissue pathophysiology associated with the progression of tissue injury from radiation exposure. Quantitative correlations with specific histological criteria were less conclusive, suggesting that more information may be needed to refine the model architecture if model parameters are to be explicitly related to the types and extent of tissue damage induced by radiation treatments. PMID- 10372147 TI - A review of parameters for the bioelectrical characterization of breast tissue. AB - In the data set collected by the authors in freshly excised breast tissue, the admittance loci generally differed from circular arcs, rendering the calculation of the usual set of parameters impossible. Alternative parameters were used for the analysis of these data. The present study consists of the definition and evaluation of a set of such parameters aimed at the characterization and differentiation of breast tissues. These parameters were defined so that their calculation does not require the fit of circular arcs to the experimental points and is independent of any equivalent circuit model. The results of the statistical analysis showed significant differences between most of the tissue groups, especially between cancerous tissue and all the other groups, which confirmed that impedance spectroscopy can be considered as potentially suitable for breast cancer detection. PMID- 10372148 TI - Ex vivo discrimination between normal and pathological tissues in human breast surgical biopsies using bioimpedance spectroscopy. AB - Ex vivo bioimpedance data measured on normal and cancerous female breast tissues are reported. They clearly show that the electrical properties of normal tissues, surrounding tissues, and carcinoma are different. These differences lie in the conductivity, in the characteristic frequency (frequency of the maximum of the imaginary part of the bioimpedance), and also in the shape of the Bode plots. Modeling using an R-S-Zcpe model is reported as well as indexes extracted from the real and imaginary parts of the bioimpedance. Even if a classification of the different types of tissues remains a difficult task and leads to much less precise diagnosis than microscopic examination, the electrical behavior of mammary tissue could be used to develop a noninvasive technique for early breast cancer detection. PMID- 10372149 TI - In vivo and in situ ischemic tissue characterization using electrical impedance spectroscopy. AB - The investigation of processes of ischemia in different organ tissues is very important for the development of methods of protection and preservation during surgical procedures. Electrical impedance spectroscopy was used to distinguish between different tissues and their degree of ischemia. We describe mathematical methods used to adjust experimental data to Cole-Cole models for one-circle and two-circle impedance loci and a study of the main parameters for representing the behavior of ischemia in time. In vivo and in situ postmortem measurements of different tissues from pigs are shown in the 100 Hz to 1 MHz range. The Cole parameters that best characterize the ischemia are R0 and fc. PMID- 10372150 TI - Dielectric properties of skeletal muscle during ischemia in the frequency range from 50 Hz to 200 MHz. AB - The complex dielectric properties of canine skeletal muscles were measured at 25 degrees C during ischemia in the frequency range from 50 Hz to 200 MHz. The dielectric spectrum of skeletal muscle shows an alpha-dispersion below 1 kHz and a beta-dispersion with a relaxation frequency of about 100 kHz. The alpha dispersion disappears between 450 and 500 min of ischemia time, the same time during which mechanical contraction was observed, and was restored later. During ischemia, the beta-dispersion is shifted continuously to higher frequencies; and at frequencies above 50 MHz, a decrease of the real part of the dielectric permittivity was measured. The dielectric loss factor decreases during ischemia at frequencies below 500 kHz, only interrupted by a short increase, coinciding with the disappearance of the alpha-dispersion. The principal processes that happen during ischemia inside the skeletal muscle tissues were studied with the help of a model especially designed to simulate membrane effects on the dielectric spectrum. The disappearance of the alpha-dispersion is explained by an increase of conductivity in the membrane of the sarcoplasmic reticulum. Shifting beta-dispersion to higher frequencies is a result of metabolically produced ions and therefore increasing conductivity of the intracellular medium. Decreasing dielectric permittivity at frequencies above 50 MHz and decreasing dielectric loss factor at low frequencies are caused by the cell edema. PMID- 10372151 TI - Quantitative analysis of impedance spectra of organs during ischemia. AB - We have developed a rapid, quantitative procedure to fit the spectra of the real and imaginary part of tissue impedance, providing characteristic parameters: time constants, their distribution, and the amplitudes of associated dispersions. Based on the time course of tissue impedance during ischemia, we have derived the evolution of characteristic parameters for both myocardial and liver tissue. The similar evolution of the distribution of time constants for myocardial and liver tissue is emphasized and discussed. PMID- 10372152 TI - Requirements for clinical use of bioelectrical impedance analysis (BIA). AB - The bioelectrical impedance analysis (BIA) method is an attractive tool for use in the clinical assessment of human body composition. Factors such as ease of use, relatively low cost, noninvasive nature, high degree of reproducibility, and safety of operation provide an impetus for the general application of this method. The preponderance of the published applications of BIA focused on applications in healthy populations and indicated a qualitative validity of the method. More recent applications have augmented the quantitative values of the BIA approach and have reported very good specificity and sensitivity. One potential limitation of the BIA approach is the reliance on regression models, derived in restricted samples of human subjects, which restricts the usefulness of the derived model in other patients who differ from the original sample in which the model was developed. Other investigational approaches that use different physical models (bioelectrical impedance spectroscopy and parallel model) have yielded successful and useful measures of human body composition in clinical studies. If BIA is to gain acceptance in clinical diagnosis and evaluation of therapeutic interventions, further efforts will be needed to ascertain more fully the validity, sensitivity, and specificity of biological parameters estimated with the new BIA approaches, and to establish the prognostic values of the BIA estimates of body composition. PMID- 10372153 TI - Study of the relation between fluid distribution change in tissue and impedance change during hemodialysis by frequency characteristics of the flowing blood. AB - Erythrocyte orientation and deformation cause differences in impedance between flowing and resting blood. Through theoretical calculation and experimental measurements, we studied the effects of these factors on blood impedance. The size and shape of the erythrocyte and the conductivity of the interior medium of the erythrocyte change when the osmotic pressure of plasma is changed. From experimental results, we obtained the following: when the size of the erythrocyte becomes larger than the normal size due to the osmotic pressure change, the beta dispersion frequency decreases and the intra- and extracellular fluid resistance increase. These experimental results corroborate that the change of tissue impedance like muscle impedance during hemodialysis is caused by the change of the fluid distribution and the change of ionic concentration of the electrolyte in tissues during hemodialysis. Also, we could estimate the relative change value of the intra- and extracellular fluid volume by the impedance method, if there were no ionic concentration change in the electrolyte. It would be very difficult to estimate the absolute change value of them because a shadow effect due to the cells depends greatly upon the shape and size of the cells and the cell concentration. PMID- 10372154 TI - Bioimpedance: is it a predictor of true water volume? AB - Bioelectrical impedance analysis (BIA) has been reported to be insensitive to changes in water volumes in individual subjects. This study was designed to investigate the effect on the intra- and extracellular resistances (Ri and Re) of the segments of subjects for whom body water was changed without significant change to the total amount of electrolyte in the respective fluids. Twelve healthy adult subjects were recruited. Ri and Re of the leg, trunk, and arm of the subjects were determined from BIA measures prior to commencement of two separate studies that involved intervention, resulting in a loss/gain of body water effected either by a sauna followed by water intake (study 1) or by ingestion (study 2). Ri and Re of the segments were also determined at a number of times following these interventions. The mean change in body water, expressed as a percentage of body weight, was 0.9% in study 1 and 1.25% in study 2. For each study, the results for each subject were normalized for each limb to the initial (prestudy) value and then the normalized results for each segment were pooled for all subjects. ANOVA of these pooled results failed to demonstrate any significant differences between the normalized mean values of Ri or Re of the segments measured through the course of each study. The failure to detect a change in Ri or Re is explained in terms of the basic theory of BIA. PMID- 10372155 TI - Fat-free mass qualitative assessment with bioelectric impedance analysis (BIA). AB - Body composition studies, when based on two-compartment volumetric estimates, can hardly assess nutritional states. Phase-sensitive impedance analysis can be used to reflect directly the proportions between intra- and extracellular spaces (ECM/BCM), which is one of the most sensitive indexes of malnutrition. Resistance and reactance values actually measured with BIA are referred to as the series RC model; however, due to the morphology of the FFM, which is composed of cells surrounded by interstitial fluids, in reality this should be modeled as a parallel RC circuit. A nomogram developed with series-to-parallel transformations of resistance and reactance measured with commercial BIA on controls and patients shows interesting gender and disease sensitivity and specificity. PMID- 10372156 TI - Estimation of extracellular volume by a two-frequency measurement. AB - An approach to determine intra- and extracellular conduction on the basis of Bode analysis is presented. Estimation of the ratio between intra- and extracellular conduction could be performed by phase measurement only, midrange in the bandwidth of interest. An important feature is that the relation between intra- and extracellular conduction can be continuously monitored by phase measurement and no curve fitting whatsoever is required. Based on a two-frequency measurement determining Re at 4 kHz and phi max at 64 kHz, it proved possible to estimate extracellular volume (ECV) in 23 patients. Reference values on ECV were determined by sodium bromide. The results show a good correlation (r = 0.90) with the reference method. The average error of ECV estimation was -3.6% (SD 8.4). PMID- 10372157 TI - The RXc graph in evaluating and monitoring fluid balance in patients with liver cirrhosis. AB - A recent study, using height-standardized resistance (R/H) and reactance (Xc/H) and assuming a bivariate distribution, has proposed the "RXc graph". We applied this new approach for patients with chronic liver disease in differentiating various degrees of fluid unbalance. Our data showed that a 95% confidence ellipse of patients with chronic hepatitis (CH) overlapped that of healthy control subjects (CONTR), while those of patients with liver cirrhosis (CIR), patients with cirrhosis and ascites (ACIR), and patients with cirrhosis, edemas, and ascites (AECIR) were clearly different for both genders. A progressively shorter mean impedance vector proportional to the stage of liver disease and to the degree of fluid unbalance was found. The lower half of the 50% tolerance ellipse for the healthy population proved to be a threshold for cirrhotics, while almost all the subjects with clinically detectable edema fell outside this limit. The RXc graph was shown to be useful in monitoring the treatment of fluid unbalance and for the immediate selection of patients in whom BIA can precisely assess body composition. PMID- 10372158 TI - Measurement of leg arterial compliance of normal subjects and diabetics using impedance plethysmography. AB - In this study, compliance, a mechanical characteristic of the lower leg arteries, was measured noninvasively. Changes in blood volume and pressure were measured using impedance plethysmography and a mercury sphygmomanometer, respectively. Compliance was calculated by dividing the change in blood volume by the change in pulse pressure (systolic-diastolic pressure). Subjects were 24 asymptomatic persons ranging from 30 to 58 years and 14 diabetics ranging from 41 to 59 years. Peak compliance, mean pressure, and systolic pressure were statistically analyzed using a t test between the asymptomatic and diabetic groups. The average peak compliance of the asymptomatic and diabetic groups was measured as 2.79 and 1.82 microL/mmHg/cm, respectively, and these were significantly different (p < 0.01). It was also found that compliance is a better parameter in differentiating vascular disease than the mean or systolic blood pressure. PMID- 10372159 TI - A meta-analysis of published studies concerning the validity of thoracic impedance cardiography. AB - Our aim was to provide a meta-analysis of the literature concerning the validation of thoracic impedance cardiography (TIC) and to explain variations in reported results from differences in the studies. One hundred fifty-four studies (164 Fisher's Z-transformed correlation coefficients) comparing measurements of cardiac output or related parameters from TIC and a reference method were analyzed. Papers were classified according to differences in TIC methodology, reference method, and subject characteristics. Pooling using the random-effects method yielded an overall correlation of r = 0.82 (95% confidence interval: 0.80 0.84). ANOVA revealed a significant influence of the reference method and the subject characteristics on the correlation coefficient. In cardiac patients, the correlation was significantly decreased. No influence of the applied TIC methodology was found. CONCLUSION: TIC might be useful for trend analysis of different groups of patients. However, since the reference method was of significant influence, differences between TIC and the reference method are incorrectly attributed to TIC alone. PMID- 10372161 TI - Lead field theoretical approach in bioimpedance measurements: towards more controlled measurement sensitivity. AB - This study was conducted to demonstrate the potentiality of lead field theoretical approach in analyzing bioimpedance (BI) measurements. Anatomically accurate computer models and the lead field theory were used to develop BI measurement configurations capable of detecting more localized BI changes in the human body. The methods were applied to assess the measurement properties of conventional impedance cardiography (ICG) and such BI measurement configurations as can be derived using (i) the 12-lead electrocardiography (ECG) and (ii) the international 10-20 electroencephalography (EEG) electrode systems. Information as to how various electrode configurations are sensitive to detecting conductivity changes in different tissues and organs was thus obtained. Theoretical results with the 12-lead system suggested that, compared to conventional ICGs, significantly more selective ICG configurations can be derived for cardiovascular structures. In addition to theoretical investigations, clinical test measurements were made with the 12-lead system to establish whether characteristic waveforms are available. Sensitivity distributions obtained with the 10-20 electrode system give promise of the possibility of monitoring noninvasively cerebrospinal fluid (CSF) impedance changes related to impending epileptic seizures. PMID- 10372160 TI - Towards a theoretical understanding of stroke volume estimation with impedance cardiography. AB - In electrical impedance cardiography, Kubicek's formula is often used to measure stroke volume from thoracic impedance variations synchronously to heart activity. To calculate stroke volume from impedance variations, the so-called outflow problem should be adequately solved. This outflow problem refers to the joint causes of impedance change due to blood entering the aorta from the heart, as well as blood leaving the aorta due to arterial runoff. The aim of this study was to investigate the Kubicek formula as a solution of the outflow problem. Kubicek's formula was theoretically investigated using a simple model of the volume-conducting properties of the thorax (two-cylinder model), as well as the hemodynamics of the systemic circulation (three-element "windkessel" model). The mathematical analysis showed that the outflow problem was not solved by the Kubicek formula. Moreover, this theoretical result was experimentally confirmed. PMID- 10372162 TI - Impedance stroke volume compared with dye and electromagnetic flowmeter values during drug-induced inotropic and vascular changes in dogs. AB - Stroke volumes measured by impedance were compared with values obtained by dye dilution and an electromagnetic flowmeter (EMF) on 14 dogs during drug-induced changes in cardiac contraction strength and peripheral resistance changes. Grouping all data for a total of 305 points showed correlations between dye and EMF, dye and impedance, and EMF and impedance of 0.89, 0.68, and 0.72, respectively. Correlations for individual dogs between dye and EMF, dye and impedance, and EMF and impedance ranged from 0.60 to 0.99, -0.39 to 0.96, and 0.26 to 0.89, respectively. These data suggest that the use of impedance cardiac output measurements to make treatment decisions about individual patients could result in serious error. PMID- 10372163 TI - A comparison of bioimpedance and echocardiography in measuring systolic heart function in cardiac patients. AB - To investigate the ability of bioimpedance cardiography to assess left ventricular systolic function in comparison with known echocardiographic parameters and to establish the most informative bioimpedance parameter, 28 cardiac patients were submitted to simultaneous echocardiography and bioimpedance cardiography. Bioimpedance systolic time ratio, Heather index, acceleration index, and index of contractility were compared with echocardiographically obtained left ventricular dimensions, 2D left ventricular ejection fraction, fractional shortening, and mean velocity of circumferential shortening. The systolic time ratio and Heather index correlated significantly well with, respectively, 2D ejection fraction (r = -0.73) and, respectively, fractional shortening (r = 0.69). The systolic time ratio was the best parameter in recognizing impaired left ventricular systolic function (F = 12.6) in comparison with the Heather index (F = 6.5). This study demonstrates the applicability of bioimpedance cardiography in assessing left ventricular systolic function similar to echocardiography in clinical cardiology. PMID- 10372164 TI - Thoracic bioimpedance as a basis for pacing control. AB - Periodic variations of the thoracic bioimpedance due to breathing and heartbeating carry confidential information that is used for pacing rate control in rate-adaptive pacemakers. The respiratory parameters--the respiration rate and tidal volume--are detected from the filtered breathing signal component (0.1 to 1.0 Hz), and are used for fuzzy feed-forward adaptive control of the pacing rate to meet the needs of the organism. The cardiac parameters--the actual heart rate and stroke volume--are measured from the heartbeating signal component (1.0 to 3.0 Hz) and are proposed for feedback correction of the feed-forward control to meet the heart's ability. The problems of electrical bioimpedance measurement and design of the rate-adaptive pacemaker, wherein the intracardiac impedance is used as the main information source for pacing control, are discussed in this paper. PMID- 10372165 TI - Evaluation of systolic performance by automated impedance cardiography. AB - Impedance cardiography (ICG) is a noninvasive method for evaluating cardiac function. Left ventricular stroke volume (SV) is the basic hemodynamic parameter derived from thoracic bioimpedance curves. Issues of our study were to investigate the diagnostic value of other indices of left ventricular systolic performance, such as ejection fraction (EF), index of contractility (IC), peak flow index (PFI), and acceleration index (ACI), which can also be calculated by ICG. Forty patients (PTS) with suspected coronary artery disease (CAD) were monitored by automated ICG during pharmacologic stress testing with dobutamine. All PTS underwent subsequent cardiac catheterization. In PTS with single vessel disease, the dobutamine-induced changes of SV, EF, IC, PFI, and ACI were comparable to those of PTS without CAD. In PTS with multivessel disease, the impaired systolic performance during dobutamine stimulation could be clearly demonstrated. We conclude that automated ICG is a useful method for monitoring SV and other indices of left ventricular systolic performance for detecting PTS with ischemic left ventricular dysfunction during cardiovascular stress. PMID- 10372166 TI - Impact of cardiovascular reactions using the impedance cardiography method in borderline hypertension. AB - This paper evaluates the hemodynamics of 30 young men, aged 17-19 years, with borderline hypertension (BHT) and 29 normotensive (NT) patients within the same age range. The study has been carried out using the impedance cardiography method at rest and under passive orthostatic test, cold test, and hyperventilation test. In the BHT patients, the following features have been observed: increased values of the cardiac index (CI), stroke volume index (SVI), mean blood pressure (MBP), and left ventricle work index (LVWI), as well as the accompanying normal values of the total peripheral resistance index (TPRI). The reactions of the cardiovascular systems during the functional tests are similar in both tested groups; however, they are most clearly distinct in the BHT patients, where the differences are statistically significant. This may support the argument that the activity of the adrenergic system in this group in intensified. PMID- 10372167 TI - Stroke volume variability--cardiovascular response to orthostatic maneuver in patients with coronary artery diseases. AB - The dynamics of cardiovascular responses to postural stress have not been fully recognized. To determine whether coronary artery bypass grafting (CABG) has any effect on stroke volume variability (SVV), the power spectrum components of SVV were measured in 60 patients before and at 6 weeks after CABG. Stroke volume was assessed by means of the thoracic bioimpedance method. The thoracic impedance cardiogram and ECG were recorded in the supine and standing positions with controlled breathing rate (0.25 Hz) during 10-minute periods. The analysis of SVV was done by means of the autoregressive method. The total power, the power in the low-frequency band LFSV (0.05-0.15 Hz), the power in the high-frequency band HFSV (0.15-0.5 Hz), and the LFSV/HFSV ratio were analyzed. Before CABG, we did not notice any significant changes in the stroke volume spectral power indices. After CABG, all spectral indices were significantly decreased in the standing position. PMID- 10372168 TI - Assessment of left ventricular systolic function and diastolic time intervals by the bioimpedance polyrheocardiographic system. AB - In order to detect left ventricular systolic function and diastolic time intervals using a new improved bioimpedance polyrheocardiographic system (BPCS), 110 healthy subjects and 128 patients with myocardial infarction were examined. Twenty-four simultaneous measurements of cardiac output by thermodilution and BPCS were performed in 11 patients with complicated acute left ventricular failure. Studies demonstrated a high degree of correlation (r = 0.91, p < 0.001). The correlation between the methods of using signals of the second derivative and the subtracted first derivative waveform of BPCS and continuous-wave Doppler echocardiography of systolic and diastolic time intervals was studies in 51 patients. The methods were closely correlated, especially with respect to left ventricular ejection time (r = 0.95), isovolumic relaxation time (r = 0.85), time to peak filling (r = 0.90), and deceleration of rapid filling time (r = 0.91). New bioimpedance hemodynamic parameters such as peak volume acceleration of ejection (PVAE) and peak power ejection (PPE) in patients with heart failure (NYHA Class I-III) were studied. Significant reductions of PVAE and PPE in groups of patients with marked progression of heart failure were noted. These results have demonstrated that BPCS is a noninvasive, simple accurate method of assessment of left ventricular systolic function and diastolic time intervals. PMID- 10372169 TI - In vivo ac impedance spectroscopy of human skin. Theory and problems in monitoring of passive percutaneous drug delivery. AB - The use of impedance spectroscopy to evaluate transdermal drug delivery is discussed and new techniques and protocols are suggested to avoid or minimize potential problems. A novel multichannel impedance analyzer, exploiting the advantages of the "three-electrode" configuration, was employed to measure the effects of differing topically applied concentrations of the percutaneous local anesthetic amethocaine on the electrical properties of the treated skin sites. Each measured impedance spectrum was modeled by an equivalent circuit consisting of a resistor in series with the parallel combination of a pseudocapacitance and a resistor. Due to differences in skin sites and to the finite times taken to apply each electrode, it was difficult to satisfactorily compare and contrast the results obtained from adjacent skin sites. Normalization of data highlighted differences in relative impedance changes and aided the meaningful comparison of treated skin sites. PMID- 10372170 TI - On assessment of skin reactivity using electrical impedance. AB - Pathophysiological events in biological tissue are characterized by a shift in electrical impedance spectra of the tissue under study. In this paper, techniques based on electrical impedance are reviewed with emphasis on their possible role in evaluating the skin reactivity of an individual, including results from impedance measurement studies on patients with allergic contact reactions, wheals, tuberculin tests, and irritant contact reactions and on an appropriate number of controls. The results show that, compared to relevant controls, at different types of experimental cutaneous reactions, both of allergic and irritant type, statistically significant changes of the impedance parameters have been detected. Each reaction type had a specific impedance index pattern. Data up to now indicate that the improved impedance technique offers not only a noninvasive alternative for characterization and perhaps differentiation between the skin responses induced by either an allergen or an irritant, but also a capability to distinguish responses induced by chemically different irritants. PMID- 10372171 TI - Electrical bioimpedance related to structural differences and reactions in skin and oral mucosa. AB - Electrical bioimpedance can reflect structural and chemical changes of the skin and the oral mucosa in the beta-dispersion frequency range. From our measured multifrequency data set, four physically distinct indices have been formulated to distinguish the electrical properties for different anatomical locations and to detect different reactions and conditions of the skin and the oral mucosa. In comparison with the skin, the differences for various anatomical regions were greater in the oral cavity, which showed as well a different impedance pattern after irritant responses. We conclude that the impedance technique is able to classify and quantify different responses and conditions, preferably by using contralateral reference sites, or following a site in time; however, mapping of baseline properties facilitates the use of the method even if a large part of the skin or the oral mucosa is involved. The method has the potential of becoming a diagnostic decision support tool. PMID- 10372172 TI - Stress action on biological tissue and tissue models detected by the Py value. AB - The Py value, a fast measurable combination of the conductivity at the corner frequencies of the beta-dispersion, is a measure of the relative cell volume concentration in tissue. In many cases, if the biological object is stressed, for instance, by mechanical deformation, shortage of oxygen, electric field strength, temperature rise, or ischemia, Py increases. Depending on the object and the kind of stress, Py plateaus for minutes up to hours and then it decreases continuously. Values of passive electrical parameters of biological tissues are often given without information about the time following a stimulation or stress situation, for example, death, surgery, field application, etc. However, since the passive electrical properties change with time, information about their history, for example, time after death, should be given. PMID- 10372173 TI - From concept to market in industrial impedance applications. AB - This paper discusses some of the technical aspects of converting a laboratory idea into a commercial product. The example used is the development of the Aber Instruments Biomass Monitor, which is now used worldwide in industry for the pitching of yeast in breweries and for biomass measurements in the pharmaceutical industry. Although the issues raised will relate to instrumentation in a production environment, many of the themes will be equally applicable to medical instruments. PMID- 10372174 TI - Orientation and deformation of erythrocytes in flowing blood. AB - The effects of flow on the changes of electrical resistivity and light-scattering characteristics of blood are experimentally and theoretically discussed. Studies indicate that most erythrocytes deform and orient themselves in the flow direction when blood flows in a conduit. Such oriented blood shows anisotropic properties. Anisotropic electrical resistivity of flowing blood is measured in three rectangular directions with a measurement cell of coaxial cylindrical type. From these experimental results, the orientation and deformation of erythrocytes are discussed. The orientation ratio and the deformation are calculated using a simplified spheroidal model of an erythrocyte. Calculated results show that the fractions of erythrocytes with their short axis parallel to each direction and the equivalent axis ratio for a simplified spheroidal model change with the shear rate of flow. PMID- 10372175 TI - On the limits of ellipsoidal models when analyzing dielectric behavior of living cells. Emphasis on red blood cells. AB - The dielectric behavior of red blood cells is simulated by taking into account the real shape (consistent with microscopic observations) and the ellipsoids (prolate and oblate spheroids) having the same surface and volume. We have pointed out that the spectra of the imaginary versus the real part of polarizability, which can be directly derived from the measured data, provide quantitative insight to cell morphology. We emphasize that ellipsoidal approximation is fairly good for random oriented cells, but rather poor whenever oriented cells are measured. This fact is assumed to be the reason for the differences between the reported parameters derived from measurements on single cells and those from observations on (random oriented) cells in suspension. PMID- 10372176 TI - Electrical impedance tomography study of biological processes in a single cell. AB - An in vivo electrical impedance tomography (EIT) study of single plant cells of Chara corallina is reported. When these aquatic cells grow in alkaline conditions, proton-translocating ATP synthases in the plasma membrane operate in reverse, utilizing ATP to translocate protons against an electrochemical gradient to the periplasm and creating localized acidic regions along the cell's cylindrical surface. These acidic regions, which appear as radial bands, approximately 5 mm long, between narrower alkaline bands, facilitate the uptake of bicarbonate, the plant's source of inorganic carbon for photosynthesis in the carbon dioxide-depleted alkaline conditions. Our EIT study of cell ultrastructure in the acidic and alkaline regions provides evidence that the plasma membrane is folded in localized regions (e.g., charasomes) in the acidic bands. The very low frequency capacitance dispersions were very similar to those of double fixed charge structures. Such charge distributions are known to be present in the membrane-bound F0 portion of the ATP synthase. The theoretical dependence of the fixed-charge concentrations on pH in the proteins is shown to broadly account for the observed correlations between pH, membrane potential, conductance, and capacitance in these regions. In synthetically formed double fixed-charge membranes, electric field-induced dissociation of water into H+ and OH- occurs. This leads to the speculation that H+/OH- fluxes in ATP synthases located in the alkaline regions of Chara cells might also involve the electric field-induced dissociation of water. PMID- 10372177 TI - New light-scattering and field-trapping methods access the internal electric structure of submicron particles, like influenza viruses. AB - A variety of AC-electrokinetic field effects can be exploited for handling or electric characterization of microscopic and submicroscopic particles, like cells, organelles, supramolecular structures, and artificial colloids. Despite the fact that dielectric spectroscopy methods by AC-electrokinetics, like common impedance methods, are based on the impedance properties of the different constituents of the particles, the first methods yield higher parameter resolutions. A drawback of the electrokinetic methods was that they required microscopic observability of field-induced particle movements. New AC electrokinetic methods like electrorotational light scattering (ERLS), dielectrophoretic phase-analysis light scattering (DPALS), and dielectrophoretic field trapping (DFT) solve this problem and access the submicroscopic particle range. This paper gives an introduction to the new methods and presents measurements on influenza viruses. To develop a dielectric virus model, experiments of ERLS were combined with DFT of viruses in microstructured electric field cages. The model assumes a spherical virus with a radius of 50 nm and a single-shell dielectric structure. The shell thickness of 18 nm summarizes the dimensions of the lipid and viral surface protein layers. For this model, the conductivities of core and shell of 0.1 mS/m and 0.1 microS/m, respectively, and the relative permittivities of 30 and 80, respectively, were obtained. PMID- 10372178 TI - Biomass monitoring using impedance spectroscopy. AB - The biomass density in biotechnological processes is often determined by indirect and manual methods. Electrical impedance spectroscopy can provide online viable biomass density estimators. In this work, we present two linear estimators obtained with this technique. Four different microorganisms were measured. The detection threshold was approximately 1 g/L (dry weight) for bacteria and 0.5 g/L for yeast. Liposome suspensions were also used to validate the methods. The monitoring of the continuous growth of a yeast culture is also presented. PMID- 10372179 TI - Virtual biopsies in Barrett's esophagus using an impedance probe. AB - Preliminary results of electrical impedance measurements in squamous and columnar epithelia in rat and human tissues are presented. The aim of this work is to show the possibility of differentiating these two types of epithelia in terms of their electrical characteristics. For the measurements, we employed a 1.95-m-long, 3.2 mm-diameter, four-electrode probe designed to be used transendoscopically in the diagnosis of Barrett's esophagus (BE). BE is a condition in which the normal squamous epithelium of the esophagus is replaced by columnar epithelium of the intestinal type. This metaplasia is considered as a premalignant condition that puts patients at a 30-125-fold risk of developing adenocarcinoma of the esophagus. The diagnosis and surveillance of BE involve taking multiple biopsies, an expensive and time-consuming procedure. This study constitutes the first stage in the replacement of tissue biopsy by "virtual biopsies". PMID- 10372180 TI - Inductively coupled wideband transceiver for bioimpedance spectroscopy (IBIS). AB - Most measurement devices for bioimpedance spectroscopy are coupled to the measured object (tissue) via electrodes. At frequencies > 500 kHz, they suffer from artifacts due to stray capacitances between electrode leads as well as between the ground and object. The noninvasive measurement of the brain conductivity is hardly possible with surface electrodes. These disadvantages can be obviated by inductive coupling. The aim of this work was the development of a wideband transceiver for inductive impedance spectroscopy. In order to define its specifications, a feasibility study has been carried out with a simulation model for three different coil systems above a homogeneous conducting plate. According to simulation results, all systems render it possible to resolve conductivity changes down to 10(-3) (omega m)-1 at frequencies > 50 kHz. The transceiver electronics must then provide a resolution of > or = 1 microV and an excitation current of up to 1 A. The realized receiver matches these specifications with an S/N ratio of 22 dB at 1 microV in the frequency range of 50 kHz to 5 MHz. PMID- 10372181 TI - Magnetic induction tomography. A measuring system for biological tissues. AB - A single-channel magnetic induction system operating at 10 MHz has been constructed. The system consists of an excitation coil and a sensing coil, between which different objects can be scanned. The eddy currents induced in the object cause perturbations in the sensed magnetic field, which are measured with a phase-sensitive detector with backing off of the signal to improve sensitivity. Scans were obtained for saline solutions with conductivities ranging from 0.001 to 6 Sm-1, encompassing the range for biological tissues. The imaginary part of the perturbation in the sensed magnetic field was found to be proportional to saline conductivity, consistent with theoretical prediction, and had a constant of proportionality of -1.2% per Sm-1. A filtered back-projection algorithm was used to generate tomographic images from the scans. PMID- 10372182 TI - Magnetic impedance tomography. AB - Tissue can be characterized by its electrical impedance, especially if measurement can be extended over a range of frequencies. Recently, there has been a great deal of interest in imaging the distribution of electrical impedance through the technique of electrical impedance tomography (EIT). However, EIT has a number of practical problems relating to the placement of electrodes on the body. Such contacts are not required to collect magnetic field data around an object through which current is flowing and thus this approach may be more practical than EIT in the clinical environment. This paper describes the technique of magnetic impedance tomography (MIT), which allows reconstruction of the current distribution from magnetic field measurements. The reconstruction techniques used to generate the images and the prototype data collection system are described. Images produced using data collected from discrete and distributed current phantoms and the thorax during human respiration are presented. PMID- 10372183 TI - Evaluation of impedance technique for detecting breast carcinoma using a 2-D numerical model of the torso. AB - Previous experimental studies showed that significant changes occur in the electrical properties of breast cancer tissue compared to the surrounding normal tissue. This phenomenon motivated studies on cancer detection using electrical impedance techniques. In the present study, a two-dimensional model of the torso and a numerical method were used to investigate the changes in the potential distribution as a result of a malignant tissue present in the breast. A transverse MRI image of the woman's torso was scanned. Noise reduction and contour-following algorithms were applied to differentiate between eight compartments in the torso. The extracted tissue types were lungs, blood, ribs, bone marrow of the cord, breast fat, skin, skeletal muscle, and heart muscle. Isotropic homogeneous conductivity was assigned to each one of these compartments. The volume conductor problem was solved numerically using the finite volume method to determine the potential distribution developed due to the dipole source. Cases without and with artificially inserted malignant region with realistic sizes were examined to investigate the sensitivity of impedance techniques to detect breast cancer. Significant changes were detected in the potential distribution inside the volume conductor as a result of the realistic size of breast tumors. A linear relation was found between the surface potential in the vicinity of the tumor region and the size of the tumor. For a small malignant area of 0.22 cm2, the surface potential near the tumor region decreased only slightly from a value of 13.81 mV in the normal case to 13.67 mV (0.14 mV change; 1.0%). For a larger malignant area of 5.43 cm2, the potential decrease was more pronounced, 11.29 mV (2.52 mV change; 18.3%), indicating that realistic sizes of breast tumor result in significant changes in the surface potential. Thus, impedance techniques employed in the present study show very good promise in detecting breast cancer. PMID- 10372184 TI - Focused impedance measurement (FIM). A new technique with improved zone localization. AB - Conventional four-electrode impedance measurements (FEIM) cannot localize a zone of interest in a volume conductor. On the other hand, the recently developed electrical impedance tomography (EIT) system offers an image with reasonable resolution, but is complex and needs many electrodes. By placing two FEIM systems perpendicular to each other over a common zone at the center and combining the two results, it is possible to obtain enhanced sensitivity over this central zone. This is the basis of the proposed new method of focused impedance measurement (FIM). Sensitivity maps in both 2D and 3D show the desired improvement. A comparison of stomach-emptying studies also indicates the improvement achieved. This new method may be useful for impedance measurements of large organs like stomach, heart, and lungs. Being much simpler in comparison to EIT, multifrequency systems can be simply built for FIM. Besides, FIM may have utility in other fields like geology where impedance measurements are performed. PMID- 10372185 TI - Monitoring regional lung ventilation by functional electrical impedance tomography during assisted ventilation. AB - A new approach in discriminating the regional air volume changes in the lungs associated with either spontaneous or mechanical ventilation during assisted ventilation is presented. Impedance data are obtained by conventional electrical impedance tomography (EIT). The data are filtered in the range of either the spontaneous or the ventilator rate and processed by the functional EIT (f-EIT) evaluation technique, whereby the variation of the respective EIT data with time is determined and imaged. EIT measurements performed in an infant during synchronized intermittent mandatory ventilation were evaluated with this method and indicated that the specific local lung volume swings related to spontaneous and mechanical inhalations can be separated and imaged as tomograms. This noninvasive approach may become useful in optimizing the ventilatory pattern during advanced forms of artificial ventilation and may help the clinician in the therapy management of individual patients. PMID- 10372186 TI - Gastric emptying in patients with type I diabetes mellitus. AB - Diabetic autonomic neuropathy is a known complication of long-standing diabetes. The present study was designed to study the prevalence of asymptomatic prolonged gastric emptying (GE) in young patients with IDDM and its correlations with disease duration and autonomic nerve function. The study population included 40 poorly controlled patients, mean age 17.6 +/- 4.6 years, with a disease duration of 1-17.5 years, and 20 age- and sex-matched controls. Autonomic nerve functions were assessed by standard cardiovascular reflexes, and gastrointestinal (GI) symptoms were assessed by a detailed questionnaire. GE was assessed by electrical impedance tomography (EIT), at 2 hours after a standard semisolid meal. Mean half time gastric emptying was significantly prolonged in diabetic patients, 54.80 +/- 26.63 versus 40.37 +/- 8.62 min (p < 0.05), with a higher prevalence in the first 3 years and after 10 years of disease duration. No differences were found between diabetics and controls regarding cardiovascular tests. No correlations were found between age, GI scores, cardiovascular tests, and GE. Patients with IDDM may suffer from prolonged GE. This is not always accompanied by autonomic impairments. As impaired gastric emptying may involve poor glycemic control and early satiety, patients with difficulties in metabolic control or poor caloric intake should be studied for the possibility of delayed gastric emptying. PMID- 10372187 TI - Assessment and calibration of a low-frequency system for electrical impedance tomography (EIT), optimized for use in imaging brain function in ambulant human subjects. AB - An EIT system has been produced that has been optimized for imaging impedance changes with scalp electrodes during brain activity in ambulant subjects. It can record from 225 Hz to 65 kHz, has a small headbox on a lead 10 m long, and has software programmable electrode selection. In calibration experiments in a small cylindrical tank filled with potassium chloride solution and samples of cucumber, noise was less than 1% with averaging, and acceptable images were produced at frequencies down to 1800 Hz. This suggests that EIT can be performed at low frequencies, which are likely to give larger signals during brain activity. Future work will include trials in humans and improvement of the current source and isolation. PMID- 10372188 TI - Impedance mammograph 3D phantom studies. AB - The results obtained using the Technical University of Gdansk Electroimpedance Mammograph (TUGEM) of a 3D phantom study are presented. The TUGEM system is briefly described. The hardware contains the measurement head and DSP-based identification modules controlled by a PC computer. A specially developed reconstruction algorithm, Regulated Correction Frequency Algebraic Reconstruction Technique (RCFART), is used to obtain 3D images. To visualize results, the Advance Visualization System (AVS) is used. It allows a powerful image processing on a fast workstation or on a high-performance computer. Results of three types of 3D conductivity perturbations used in the study (aluminum, Plexiglas, and cucumber) are shown. The relative volumes of perturbations less than 2% of the measurement chamber are easily evidenced. PMID- 10372190 TI - American Society of Hypertension 14th scientific meeting. New York, New York, USA. May 20-22, 1999. Abstracts. PMID- 10372189 TI - Can we optimize electrode placement for impedance pneumography? AB - In this paper, we discuss issues involved in defining an optimum placement of four electrodes for impedance pneumography. We observed a general trend where the change in impedance (delta Z) decreased while the sensitivity (delta Z/Z) increased with distance between the drive and receive electrode pairs. However, the theoretical study indicated that delta Z/Z should decrease with distance. The scatter of points in the plots indicated that sensitivity was influenced by factors other than distance. The correlation coefficient between the theoretical and measured delta Z/Z was low, but significant. This suggested that the best electrode configuration can be derived from the theoretical data. High delta Z/Z was obtained when the drive and receive electrode pairs were placed close to the lungs and in different horizontal planes. PMID- 10372191 TI - 68th Annual meeting of the American Association of Physical Anthropologists. Columbus, Ohio, USA. April 26-May 1, 1999. Abstracts. PMID- 10372192 TI - 12th International Conference on Antiviral Research. Jerusalem, Israel, March 21 25, 1999. Abstracts. PMID- 10372193 TI - 25th Annual meeting of the European Group for Blood and Marrow Transplantation and 15th meeting of the Nurses Group. Hamburg, March 21-24, 1999. Abstracts. PMID- 10372195 TI - XXVIth European Symposium on Calcified Tissues. Maastricht, The Netherlands, 7-11 May 1999. Abstracts. PMID- 10372194 TI - British Pharmacological Society Meeting. London, United Kingdom, 6-8 January 1999. Abstracts. PMID- 10372196 TI - [Diabetes. ALFEDIAM (French Language Association for the Study of Diabetes and Metabolic Diseases. Paris, France, 30 March-4 April 1999. Abstracts]. PMID- 10372198 TI - 33rd Annual meeting of the European Society for Clinical Investigation. Milan, Italy, 8-10 April 1999. Abstracts. PMID- 10372197 TI - 23rd Annual meeting of the German Society for Cell Biology. Rostock, March 14-18, 1999. Abstracts. PMID- 10372199 TI - XIVth Congress of the European Association of Urology (EAU). Stockholm, Sweden, April 7-11, 1999. Abstracts. PMID- 10372200 TI - American Society for Gastrointestinal Endoscopy (ASGE) meeting. Orlando, Florida, USA. May 16-19, 1999. Abstracts. PMID- 10372202 TI - 72nd Annual meeting of the Japanese Pharmacological Society. Sapporo, Japan, 22 25 March 1999. Abstracts. PMID- 10372201 TI - European Federation for Immunogenetics 13th Conference. Heraklion, Crete, Greece, April 13-17, 1999. Abstracts. PMID- 10372203 TI - 22nd Annual meeting of The Society of General Internal Medicine. San Francisco, California, USA. April 29-May 1, 1999. Abstracts. PMID- 10372204 TI - 46th Annual meeting of the American College of Sports Medicine. Seattle, Washington, USA. June 2-5, 1999. Abstracts. PMID- 10372206 TI - The American Pediatric Society and The Society for Pediatric Research 1999 annual meeting. San Francisco, California, USA. May 1-4, 1999. Abstracts. PMID- 10372205 TI - New developments in the treatment of the lymphomas and leukemias. Highlights from the 40th annual meeting of the American Society of Hematology (ASH). Miami, Florida, USA. December 4-8, 1998. Abstracts. PMID- 10372207 TI - The VIIIth ESPID Conference (European Society for the Study and Prevention of Infant Death), the International Conference on Prevention of Infantile Apnea and Sudden Infant Death on the Verge of the Millenium. Jerusalem, Israel, May 30-June 3, 1999. Abstracts. PMID- 10372208 TI - 67th Annual meeting of the Swiss Society of Internal Medicine and annual Swiss Medical Societies meetings. Bale, Switzerland, 15-17 April 1999. Abstracts. PMID- 10372209 TI - 13th Annual meeting of the Associated Professional Sleep Societies. Orlando, Florida, USA. June 19-24, 1999. Abstracts. PMID- 10372210 TI - [65th Annual meeting of the German Society for Cardiology-Heart and Circulatory Research. Mannheim, 8-10 April 1999. Abstracts]. PMID- 10372211 TI - [Antibody production by phage library]. PMID- 10372212 TI - [Gene transfer into hematopoietic stem cells]. PMID- 10372213 TI - The guide to occupational therapy practice. American Occupational Therapy Association. PMID- 10372214 TI - [Opioid detoxification under anesthesia: new scientific territory or established methods?]. PMID- 10372215 TI - [Forced opioid detoxification under general anesthesia--a new challenge for anesthetists and intensive care physicians]. AB - Treatment of opioid addicts by means of competitive opioid receptor antagonists was developed at the University of Vienna in 1987 by Loimer and co-workers. They compared two withdrawal regimens: The short Opiate withdrawal using a staggered naloxone regimen and the rapid opiate detoxification during general anesthesia by means of high doses of naloxone. Based on the latter concept, various modifications have been developed world-wide using either naloxone or as an alternative, naltrexone, an antagonist available for oral administration only. However, there are considerable objections to opioid detoxification during general anasthesia. The main criticism is based an the supposedly unacceptable high risk:benefit-ratio, the higher costs, the lack of psycho-social support, and the lack of prospective studies. However, first results suggest that rapid detoxification procedures are more successful in decreasing relapse than methods which are based on psychiatric treatment alone. As sympathetic hyperfunction is common in rapid detoxification procedures using high doses of opioid receptor antagonists, it is essential to avoid severe autonomic imbalance with possible subsequent impairment of organ functions. To prevent those disturbances, general anesthesia plays an important role. So far, there is almost no information about such methods in the anesthesiological literature. In this article the clinical relevance of such methods is discussed summarizing both the available literature and our own experience and we conclude that rapid opioid detoxification under general anesthesia is a safe and efficient method to suppress withdrawal symptoms. This treatment may be of benefit in patients who particularly suffer from severe withdrawal symptoms and who have failed repeatedly to complete conventional withdrawal. PMID- 10372216 TI - [Quantification of postoperative vigilance by evoked potentials]. AB - OBJECTIVE: Patients will be discharged from the postoperative recovery room mostly on subjective clinical assessment. In this study an approach to a more quantitative judgment of postoperative vigilance is made by recording the P300 latency and neuropsychological tests. METHODS: 22 adult patients undergoing a disc operation were examined. For induction of anesthesia thiopental (4-5 mg/kg), fentanyl (0.1 mg) and a musclerelaxant (atracurium 0.4 mg/kg or succinylcholine 1 2 mg/kg after precurarisation with atracurium 5 mg) were given. Anesthesia was then continued with enflurance (1.0-1.2 MAC) in a mixture of 67% nitrous oxide in oxygen. If postoperative analgesia was needed, piritramid was injected in boli a 3-6 mg. The P300 was acoustically stimulated with an oddball-paradigm and recorded at Fz and Cz. Afterwards the latencies were measured and compared with a vigilance score composed of clinical parameters and neuropsychological tests. Recordings were done preoperatively and every 30 minutes up to 2 hours postoperatively. A correlation between P300-latencies and vigilance score was made with the coefficient of Spearman. Comparison of pre- and postoperative values was managed by using Wilcoxon test for matched pairs with Bonferroni correction. RESULTS: Immediately after operation, P300 was obtained only in 8 patients (36%). The latencies were delayed (394 +/- 35 ms versus 326 +/- 12 ms preoperatively). During follow-up patients recovered and 2 hours postoperatively only one patient had no P300. At the end of the examination period P300-latencies of most patients had not yet reached the preoperative levels. The vigilance score in parallel showed decreases immediately after the operation and increases later on. However there were discrepancies between P300 latencies and neuropsychological findings, in some cases possibly due to the sedative effects of postoperative analgetics. CONCLUSION: Recording of P300-latencies showed remarkable cognitive deficits because of subclinical anesthesia hangover even 2 hours after a routine inhalational anesthesia. It is a good quantifiable method for assessment of postoperative vigilance. In some cases P300-latency is a more sensitive parameter for vigilance phenomena than clinical and neuropsychological scores. PMID- 10372217 TI - [Prophylactic analgesia in functional endoscopic sinus surgery. Hemodynamics, surgical conditions, stress response]. AB - OBJECTIVE: Sufficient control of intraoperative bleeding in functional endoscopic sinus surgery is essential for obtaining adequate surgical results. The necessity of hypotensive anesthetic techniques is a controversial topic among anesthesiologists and ENT-surgeons. This prospective, randomized study compared N2O-supplemented intravenous anesthesia with propofol and fentanyl or sufentanil with respect to hemodynamic reactions, endocrine stress response, blood loss and surgical conditions, and recovery. METHODS: After obtaining informed consent, 32 patients undergoing endoscopic sinus procedures were anesthetized with N2O, propofol, and fentanyl or sufentanil (dosage ratio fentanyl:sufentanil = 7:1). Arterial blood pressure was measured via an arterial line, blood samples for ACTH, AVP, and cortisol were obtained pre-, intra-, and post-operatively, and a psychomotor function test was conducted pre- and postoperatively. The ENT-surgeon estimated the dryness of the surgical field on a numeric scale ranging from 1 to 5, and intraoperative blood loss was measured. RESULTS: Hemodynamic reactions to surgical simulation were blunted more sufficiently in the sufentanil group. Surgical conditions were satisfactory in all patients, but significantly better in the sufentanil group; differences in blood loss did not prove statistically significant. The endocrine stress response was efficiently blunted without significant differences between the groups. Post-operative psychomotor testing showed better recovery in the sufentanil group. CONCLUSIONS: N2O-supplemented intravenous anesthesia is suitable for functional endoscopic sinus procedure without any further need for induced hypotension; sufentanil seems to be superior in regard to hemodynamic stability, surgical conditions, and psychomotor recovery. PMID- 10372219 TI - [Qualified continuing education according to the recommendations of DGAI and OGARI. Anesthesia in gynecology and obstetrics]. PMID- 10372218 TI - [Serum level-adjusted dosage of once-daily aminoglycoside therapy in critical illness: results of a prospective study]. AB - OBJECTIVE: In critically ill patients, the adjustment of target peak and trough levels of tobramycin was investigated because aminoglycoside pharmacokinetics can be changed by multiple influences. Sufficient but not too high peak serum concentrations and low trough levels, however, should be achieved to ensure a therapeutic effect and to minimize toxicity. METHODS: 70 critically ill patients of 51 +/- 18 years were monitored daily during their aminoglycoside treatment on the intensive care unit targeting a peak of about 12 micrograms/ml 30 minutes after infusion and a trough level below 1 to 2 micrograms/ml. Dose recommendations were given daily, taking into consideration serum levels, dose predictions (Bayesian method, ABBOTTBASE), creatinine clearance and clinical findings. Creatinine clearance was estimated according to the Cockcroft-Gault formula as well as directly by the urine collection method. RESULTS: The standardized initial dose of 400 mg tobramycin led to average peak serum levels of 14.2 +/- 3.9 micrograms/ml in the patients with an apparent distribution volume of 0.345 +/- 0.074 L/kg. In 95% of the patients, the initial peak was higher than 8.5 micrograms/ml; levels higher than 20 micrograms/ml were observed in 7%, extremely low concentrations (below 5 micrograms/ml) in 2%. With individually adjusted doses between 160 and 560 mg, a mean peak of 11.5 +/- 2.7 micrograms/ml was measured subsequently. The levels amounted to 96 +/- 23% of the predicted values, deviations greater than 50% occurred in 5%. The target trough level was achieved in 99%, in less than 3% the dosing interval was extended up to 72 hours. A tobramycin clearance below 80 ml/min/1.73 m2 was associated with average 80% and 33% higher creatinine clearance values according to the Cockcroft method and the direct method, respectively. CONCLUSION: Target peak and trough aminoglycoside levels are adjustable even in critically ill patients. Reduced tobramycin clearance can be associated with normal creatinine clearance. Assuming an exact methodology, a reduced "direct" creatinine clearance, however, indicates a reduced drug clearance. PMID- 10372220 TI - [Neuromuscular blockade in the inducing period?--Con]. PMID- 10372221 TI - [Neuromuscular blockade in the induction period?--Pro]. PMID- 10372222 TI - [Ethylene glycol poisoning and brain injury--a dangerous combination]. AB - We report on a patient after brain injury additionally showing signs of ethylene glycol intoxication. CT-scan showed a subdural hematoma, which in spite of increasing neurological deficit didn't show any enlargement. Metabolic acidosis with an increased anion gap and osmolar gap led to the diagnosis of ethylene glycol intoxication. Then intensive hemodialysis and i.v. ethanol were administered and the intoxication could be treated successfully. PMID- 10372223 TI - [Incidence of disposition for malignant hyperthermia in patients with neuromuscular diseases. F. Wapplet, et al., AINS 1998;33:373-80]. PMID- 10372224 TI - Cardiovascular disease in developing countries: myths, realities, and opportunities. AB - The burden of cardiovascular disease (CVD), especially ischemic heart disease and stroke, varies remarkably between regions of the world, with declining rates in Europe, North America, and Australia/New Zealand, burgeoning epidemics in the former socialist economies and India, and relatively lower impact in developing regions such as sub-Saharan Africa. The basis for a prediction of a global CVD epidemic lies in the "epidemiologic transition," in which control of infectious, parasitic, and nutritional diseases allows most of the population to reach the ages in which CVD manifests itself. In fact, CVD is already the leading cause of death not only in developed countries but, as of the mid-1990s, in developing countries as well. A variety of myths have attempted to minimize the rationale for CVD control in developing countries. In reality, CVD affects men, not only the elderly, and the rich, but rather a broad spectrum of the population. Moreover, as a cause of disability it will be a world leader by 2020. Finally, there is evidence that the epidemic can be curtailed. Projections to the year 2020 predict an expansion of the CVD epidemic to the developing world, with CVD exceeding infectious and parasitic diseases in all regions except sub-Saharan Africa. These estimates, in fact, may be conservative, because several factors may allow multiplication of risk. In utero or early childhood deprivation, the use of disposable income for deleterious health behaviors (such as tobacco and a high fat/cholesterol diet), interactions between multiple coexisting risk factors, and the interaction between newly acquired health behaviors and genes may all inflate the risk to levels above those predicted. Efforts to control CVD should invest strategically in research to understand the prevalence of, and risks associated with, CVD risk factors, as well as in studies of new risk factors, measures to prevent or modify risk, and clinical trials to demonstrate the efficacy of these interventions. In lieu of this improved research base, a number of initiatives should go forward to prevent the dissemination of risk factors, to treat risk factors appropriately in high-risk subjects, and to develop case-management strategies shown to be both efficacious and cost effective. A global epidemic of CVD in developing countries may be inevitable unless there is a better understanding of its origins, a prediction of its magnitude, and the organization of preventive and case-management strategies early enough to control it. PMID- 10372225 TI - Effects of pimobendan on the L-type Ca2+ current and developed tension in guinea pig ventricular myocytes and papillary muscle: comparison with IBMX, milrinone, and cilostazol. AB - In this study, we compared the effects of pimobendan (PIM), a putative Ca(2+) sensitizer and phosphodiesterase (PDE) inhibitor, on the L-type Ca2+ current (ICa) of guinea-pig ventricular myocytes and contractile tension of ventricular papillary muscles with those of a nonselective PDE inhibitor, isobutylmethylxanthine (IBMX), and selective PDE-III inhibitors, that is, milrinone (MIL) and cilostazol (CIL). The efficacy (maximum attainable effect) of these drugs for increasing ICa or developed tension (DT) ranged in the order of IBMX >> MIL > PIM > CIL. This finding suggests that the positive inotropic effect of each drug is roughly proportional to its increasing effect on ICa. The additional effect of PIM (a Ca(2+)-sensitizing effect) was not identified in "intact" preparations, and the potentiating effects of PIM on the DT and ICa were virtually the same as those observed for MIL. To isolate the Ca(2+)-sensitizing effect of PIM on the DT, we studied the effects of PIM in the presence of H89, an isoquinoline derivative possessing a selective inhibitory effect on cAMP dependent protein kinase. In the absence of H89, 50 microM PIM increased the DT by 68 +/- 11% (mean +/- SE, n = 6). However, in the presence of 20 microM H89, which completely blocked the PIM-induced increase in ICa, PIM (50 microM) significantly increased the DT by 19 +/- 6% (n = 6), thereby indicating the presence of a positive inotropic effect attributable to a mechanism other than increased intracellular cAMP, that is, a Ca(2+)-sensitizing effect. The latter notion was supported by the finding that in the presence of H89 (20 microM), the PIM-induced augmentation of DT was accompanied by a prolongation of the time to 50% relaxation of contractile tension. In contrast, MIL (50 microM) and forskolin, a direct activator of adenylate cyclase (1-10 nM), did not increase DT in the presence of 20 microM H89. These results suggest that the fraction of positive inotropic effect of PIM attributable to its Ca(2+)-sensitizing effect is masked by its potent PDE-III inhibitory effect in "intact" ventricular preparations. PMID- 10372226 TI - Mibefradil, a T-type and L-type calcium channel blocker, limits infarct size through a glibenclamide-sensitive mechanism. AB - Mibefradil is a novel calcium channel blocker with activity at both L-type and T type calcium channels. There are data suggesting that this compound can protect the ischemic/reperfused myocardium in spite of the fact that there is a very low abundance of T-type calcium channels within ventricular tissue. The aims of this study were two-fold. First, we wished to study the protective effect of mibefradil on ischemia/reperfusion injury in the isolated rat heart using infarct size as the endpoint of injury. In this respect, we compared mibefradil with amlodipine, a well-known and potent L-type calcium channel blocker, and with ischemic preconditioning, an intervention known to reduce infarct size consistently. Secondly, we investigated the possible mechanisms through which protection was achieved. For this second purpose, we examined the effects on protection of glibenclamide (an ATP-dependent K+ channel blocker) and chelerythrine (a protein kinase C inhibitor). Isolated rat hearts were perfused in the Langendorff mode at constant pressure. Control, mibefradil-treated (0.3 microM), mibefradil plus glibenclamide (50 microM), and mibefradil plus chelerythrine (10 microM) treated hearts underwent 35 minutes regional ischemia followed by 120 minutes reperfusion. At the end of the experiments, infarct size was determined with triphenyltetrazolium chloride and was expressed as a percentage of the ischemic risk zone (I/R%). A significant reduction in infarct size with mibefradil treatment was observed (I/R 11.1 +/- 2.1% vs. 35.5 +/- 3.1% in controls). This was comparable with the infarct reduction seen with two 5 minute cycles of ischemic preconditioning (17.7 +/- 2.5%). Amlodipine 0.1 microM, a concentration that caused equivalent coronary vasodilatation as that produced by mibefradil treatment, had no significant effect on infarct size (I/R 29.7 +/- 3.5%). The protective effect of mibefradil was not significantly modified by the presence of the PKC inhibitor chelerythrine 10 microM (I/R 19.1 +/- 4.9%) but was abolished when glibenclamide 50 microM was coadministered with mibefradil prior to ischemia (I/R 28.1 +/- 4.7%). Neither chlelerythrine nor glibenclamide alone had any influence on infarct size. We conclude from these data that mibefradil, unlike amlodipine, markedly reduces infarct size in the rat isolated heart. This protection is sensitive to inhibition by glibenclamide, suggesting that KATP channel opening may be an important additional and novel mechanism of mibefradil's action. PMID- 10372227 TI - Beta-blocker selectivity at cloned human beta 1- and beta 2-adrenergic receptors. AB - The ratio between the affinities of beta-blockers for the beta2- and beta1 receptors is often used to predict the cardioselectivity and the potential consequences of blocking beta2-receptor-mediated effects of adrenergic receptor blockers. These ratios have been traditionally determined using various in vitro models of beta2 and beta1-receptor antagonist activity, including isolated organ preparations and radioligand binding in tissues from various species. The data from these studies, while useful, are complicated by the use of different preparations, techniques, and nonhuman models. Recombinant cell lines expressing human beta2 and beta1 receptors have been developed, allowing for the direct comparison of the affinities of the beta-blockers for the beta2 and beta1 receptors under identical conditions, and allowing a precise determination of the beta1-receptor selectivity of the beta-blockers. Bisoprolol, atenolol, propranolol, betaxolol, metoprolol, carvedilol, and ICI 118, 551 were compared for their beta-receptor selectivity using membranes prepared from recombinant cells selectively expressing human beta2 and beta1 receptors. Bisoprolol was found to have the highest selectivity for the beta1 receptor, displaying a beta2/beta1 ratio of 19 (a 19-fold higher affinity for the beta3 receptor than for the beta2 receptor). Atenolol, metoprolol, and betaxolol displayed lower, selectivity for the beta1 receptor, whereas propranolol and carvedilol displayed no significant beta-adrenergic selectivity. ICI 118,55 was selective for the beta2 receptor. The equilibrium dissociation constants of the beta-blockers for the beta1 and beta2 receptors were generally similar to previously reported values. The affinity ratios were also generally similar to previously reported values. PMID- 10372229 TI - The search for the hibernating myocardium--have we reached the limit? PMID- 10372228 TI - Similar risk reduction of death of extended-release metoprolol once daily and immediate-release metoprolol twice daily during 5 years after myocardial infarction. AB - The pooled results from five placebo-controlled postinfarction studies with metoprolol have shown a significant reduction in total mortality. All five studies used immediate-release metoprolol twice daily. An extended-release formulation of metoprolol for once-daily use has since been developed. The aim of the present study was to compare the two different forms of metoprolol with regard to the risk reduction of death for 5 years postinfarction and to analyze whether treatment with the beta-blocker metoprolol is associated with a reduced mortality after the introduction of modern therapies such as thrombolysis, aspirin, and ACE inhibitors. All patients discharged after an acute myocardial infarction (AMI) from Sahlgrenska University Hospital (SU) during 1986-1987 (n = 740, Period I) and during 1990-1991 (n = 1446, Period II) from both SU and Ostra Hospital, Goteborg, Sweden, were included in the study. During Period I, 56% were prescribed immediate-release metoprolol compared with 61% prescribed extended release metoprolol during Period II. Immediate-release metoprolol was not available for outpatient use during Period II. In a multivariate analysis, all variables significantly associated with either increased or decreased postinfarction mortality during Periods I and II (univariate analysis of patient characteristics, medical history, complications during the AMI medication at discharge) studied were with Cox's proportional hazards model. Treatment with immediate-release metoprolol was significantly associated with reduced mortality over 5 years during Period I (relative risk reduction for total mortality, -34%, P = 0.003; 95% CI for RR, 0.51-0.87), and treatment with extended-release metoprolol was significantly associated with reduced mortality during Period II ( 34%, P < 0.0001; 95% CI for RR, 0.53-0.82). Thrombolysis and the use of aspirin and ACE inhibitors were more frequently used during Period II. The results showed that postinfarction treatment with extended-release metoprolol given once daily was associated with a similar risk reduction of death over 5 years as immediate release metoprolol given twice daily. The data, furthermore, indicate that the beta-blocker metoprolol is associated with a reduced risk of death after the introduction of modern therapy such as thrombolysis, aspirin, and ACE inhibitors. PMID- 10372230 TI - Effect of trimetazidine on late potentials after acute myocardial infarction. AB - The purpose of this study was to evaluate the effect of trimetazidine on late potentials in patients with acute myocardial infarction. A total of 60 patients (52 males, mean age 55 +/- 2 years, and 8 females, mean age 54 +/- 1.8 years) with the diagnosis of acute myocardial infarction were included in this study. The study was designed as a randomized, double-blinded, and placebo-controlled trial. Signal-averaged electrocardiography and echocardiography were performed during the first 2 days of acute myocardial infarction and were repeated between days of 8 and 15 (mean 11). Patients were treated with trimetazidine (n = 30) or placebo (n = 30). In the placebo group, the total filtered QRS duration and low amplitude terminal signal duration increased (from 102.7 +/- 1.8 ms to 113.3 +/- 1.8 ms, and from 32.2 +/- 0.9 ms to 38.3 +/- 1.1 ms; P < 0.001), the root mean square voltage of the terminal 40 ms of the QRS decreased (from 28.6 +/- 2.1 microV to 21.4 +/- 1.3 microV; P < 0.001), and the incidence of late potentials increased (from 30% to 46%; P < 0.01) significantly. In the trimetazidine group, these measurements were a decrease from 102.9 +/- 1.9 ms to 100 +/- 2.0 ms (NS), an increase from 31.6 +/- 0.9 ms to 32.5 +/- 0.9 ms (NS), a decrease 9.3 +/- 2.0 microV to 27.3 +/- 1.8 microV (P < 0.01), and a decrease from 33% to 30% (NS), respectively. The ejection fraction was 47.1 +/- 1.3% to 50.8 +/- 1.2% in the placebo group (P = 0.05), and 48.1 +/- 1.1% to 53.4 +/- 1.2% (P < 0.01) in the trimetazidine group. It is concluded that trimetazidine reduces late potentials after acute myocardial infarction without changing blood pressure and heart rate. PMID- 10372231 TI - Chronic treatment with an ACE inhibitor, temocapril, lowers the threshold for the infarct size-limiting effect of ischemic preconditioning. AB - This study examined whether chronic inhibition of the angiotensin-converting enzyme (ACE) lowers the threshold of preconditioning (PC) and potentiates cardio protection of subthreshold PC. Rabbits were orally administered either 0.5 mg/kg/day temocapril or placebo for 14 days and were randomly subjected to subthreshold PC (i.e., PC with 2 minutes ischemia/5 minutes reperfusion) or no PC before a 30-minute coronary artery occlusion and a 3-hour reperfusion. The size of the infarct was determined by tetrazolium staining and was expressed as the percent of the area at risk (%IS/AR). At the end of the experiment, the lungs were sampled for a tissue ACE activity assay. There was no significant difference in %IS/AR among the rabbits given placebo alone, placebo plus 2 minutes PC, and temocapril alone (%IS/AR = 59.5 +/- 5.8%, 43.6 +/- 3.7%, and 56.7 +/- 7.1%, respectively). However, %IS/AR was significantly reduced in the group that received temocapril and 2 minutes PC before infarction (%IS/AR = 35.4 +/- 4.8%, P < 0.05 vs. placebo control and temocapril control). The lung tissue ACE activity did not differ in the placebo-treated rabbits with and without PC (12.6 +/- 2.8 vs. 13.7 +/- 2.0 nmol/g protein/min) but was suppressed in temocapril-treated rabbits despite 2 minutes PC (0.9 +/- 0.1 vs. 1.5 +/- 0.2 nmol/g protein/min, both P < 0.05 vs. placebo-treated groups). The present results suggest that chronic inhibition of ACE is beneficial for potentiating the anti-infarct effect of subthreshold PC. PMID- 10372232 TI - Selective tyrosine kinase inhibitor for the platelet-derived growth factor receptor in vitro inhibits smooth muscle cell proliferation after reinjury of arterial intima in vivo. AB - The long-term success of coronary angioplasty is limited by restonosis. This study was undertaken to investigate whether and to what extent the enhanced proliferative response observed in a balloon reinjury model of rat aorta is regulated by the PDGF receptor (PDGF-R). Balloon injury was performed to 14-day old pre-existing neointimal lesion in rat aorta. PDGF receptor and ligand immunoreactivity were measured at several time points after the first and second injury, and PDGF-R signaling was blocked with a selective inhibitor of PDGF-R tyrosine kinase. In the neointima, after repeated injury, upregulation of PDGF-AA was seen to coincide with a prompt proliferative response of smooth muscle cells (SMC). Administration of the PDGF-R tyrosine kinase inhibitor in vivo, tested and found to inhibit the proliferation of SMC induced by PDGF-AA and PDGF-BB, but not by IGF-1, EGF, or bFGF, resulted in a 60% reduction in the absolute number and percentage of BrdU + cells after the second balloon injury to pre-existing neointima, but had no significant effect on proliferation after the first injury. Endpoint lesion area was reduced by 50% in the treated group at 14 days after the second injury. The results suggest that systemic administration of a tyrosine kinase inhibitor specific for the PDGF-R can be useful in the prevention of restenosis. PMID- 10372233 TI - Hyponatremia and amlodipine therapy. PMID- 10372234 TI - Therapeutic organizer (TH.OR.): a new tool in critical patient management. PMID- 10372235 TI - Impaired glucose tolerance. Why is it not a disease? PMID- 10372236 TI - Treating obesity in type 2 diabetes. Calories, composition, and control. PMID- 10372237 TI - Effect of energy restriction, weight loss, and diet composition on plasma lipids and glucose in patients with type 2 diabetes. AB - OBJECTIVE: To determine the optimal diet for improving glucose and lipid profiles in obese patients with type 2 diabetes during moderate energy restriction. RESEARCH DESIGN AND METHODS: A total of 35 free-living obese patients with type 2 diabetes were assigned to one of three 1,600 kcal/day diets for 12 weeks. The diets were high carbohydrate (10% fat, 4% saturated), high monounsaturated fat (MUFA) (32% fat, 7% saturated), or high saturated fat (SFA) (32% fat, 17% saturated). RESULTS: Diet composition did not affect the magnitude of weight loss, with subjects losing an average of 6.6 +/- 0.9 kg. Energy restriction and weight loss resulted in reductions in fasting plasma glucose (-14%), insulin ( 27%), GHb (-14%), and systolic (-7%) and diastolic blood pressure (-10%) levels and the glucose response area (-17%) independent of diet composition. Diet composition did affect the lipoprotein profile. LDL was 10% and 17% lower with the high-carbohydrate and high-MUFA diets, respectively, whereas no change was observed with the high-SFA diet (P < 0.001 for effect of diet). HDL was transiently reduced on the high-carbohydrate diet at weeks 1, 4, and 8, whereas higher fat consumption maintained these levels. The total cholesterol:HDL ratio, although significantly reduced on the high-MUFA diet (P < 0.01), was not different from the other two diets after adjustment for baseline differences. CONCLUSIONS: Energy restriction, independent of diet composition, improves glycemic control; however, reducing SFA intake by replacing SFA with carbohydrate or MUFA reduces LDL maximally during weight loss and to a greater degree than has been shown in weight-stable studies. PMID- 10372238 TI - Pharmacologic induction of weight loss to treat type 2 diabetes. AB - OBJECTIVE: Most individuals with type 2 diabetes are overweight, and weight loss for them is an important therapeutic objective. However, usual weight-loss strategies have generally not produced sustained weight loss. Pharmacologic agents to assist weight loss might be useful, but no long-term data on their effectiveness and safety in patients with type 2 diabetes are available. We therefore initiated a 2-year placebo-controlled trial of the weight-loss medications fenfluramine and phentermine in type 2 diabetic subjects. RESEARCH DESIGN AND METHODS: A total of 44 overweight (> 120% ideal body weight) subjects with type 2 diabetes were enrolled in a randomized, placebo-controlled, double blind trial of fenfluramine and phentermine. All subjects received intensive nutrition counseling, an exercise prescription, and instruction in behavior modification. Subjects were randomly assigned to 20 mg fenfluramine three times a day and 37.5 mg phentermine daily (n = 23) or dual placebos (n = 21). Diabetes medications were adjusted as necessary to achieve glycemic goals. Changes in weight, glycemia, lipemia, and blood pressure were assessed every 2 months, as were adverse events. In September 1997, when fenfluramine was withdrawn from the U.S. market, fenfluramine was stopped in all subjects. Thus the length of drug treatment varied, but 16 subjects (8 in each group) reached 12 months of treatment. Only data obtained before the withdrawal of fenfluramine are included in this report. RESULTS: A study termination, diabetes medications had been reduced in 1 subject in the placebo group (5%) and 11 subjects in the drug treatment group (52%) (P = 0.005). Drug treatment resulted in significant reductions in body weight, BMI, and HbA1c at all time points through 6 months. Changes in weight at 6 months were -2.7 +/- 1.4 kg (mean +/- SEM) with placebo treatment and -9.6 +/- 1.5 kg with drug treatment (P = 0.003). Even though more subjects in the drug treatment group required reductions in diabetes medications, at 6 months, changes in HbA1c were -0.3 +/- 0.2% with placebo treatment and -1.6 +/- 0.3% with drug treatment (P = 0.002). Fasting plasma glucose and triglycerides were significantly reduced at some time points with drug treatment. No serious adverse events attributable to study medications were observed. CONCLUSIONS: Premature study termination decreased the power of our study at later time points. However, our data suggest that weight loss medications are an effective treatment for type 2 diabetes during active weight loss. Whether the benefit persists after weight loss has stopped remains to be determined. PMID- 10372239 TI - How valid is fasting plasma glucose as a parameter of glycemic control in non insulin-using patients with type 2 diabetes? AB - OBJECTIVE: To assess the value of fasting blood glucose as a parameter for glycemic control in type 2 diabetic patients not using insulin. RESEARCH DESIGN AND METHODS: In 1,020 type 2 diabetic patients treated with diet or oral hypoglycemic agents (OHAs), measurements of fasting plasma glucose (FPG) and HbA1c were taken. In 617 patients, the measurement could be repeated after 3 months. Cross-sectional correlation coefficients were calculated for the association between HbA1c and FPG. Receiver-operating characteristic (ROC)-curve analyses were applied to examine the performance of FPG as a diagnostic test for HbA1c. Longitudinally, the change in FPG was compared with the change in HbA1c, with both correlation measures and ROC curve analyses. RESULTS: Correlation coefficients between HbA1c and FPG and between FPG change and HbA1c change were 0.77 and 0.65, respectively. ROC curve analysis showed that HbA1c is difficult to predict from FPG values: 66% of the patients with good HbA1c (< 7.0%) were identified as such by FPG values < 7.8 mmol/l. As a test for HbA1c change, FPG change performed moderately: the highest combined values of sensitivity and specificity (87.7 and 57%, respectively) were reached at a cutoff point of zero in the range of FPG change values. CONCLUSIONS: FPG and HbA1c values that do not correspond are not rare in type 2 diabetic patients on diet or OHA treatment. HbA1c is difficult to predict from FPG values, and even more difficult is the prediction of HbA1c changes from FPG changes. PMID- 10372240 TI - Effect of troglitazone on body fat distribution in type 2 diabetic patients. AB - OBJECTIVE: Troglitazone was recently reported to specifically promote the differentiation of pre-adipocytes into adipocytes in vitro in subcutaneous fat only, indicating a relation to insulin-resistance-improving action of troglitazone. To expand on this finding, we investigated at the clinical level how long-term administration of troglitazone influences the body fat distribution in type 2 diabetic patients. RESEARCH DESIGN AND METHODS: Troglitazone (400 mg/day) was administered for 6 months to 30 type 2 diabetic patients whose glycemic control was poor. A total of 18 patients received diet therapy alone (in the single-treatment group, BMI 26.0 +/- 4.6, HbA1c 8.2 +/- 1.7%), and 12 patients concomitantly received glibenclamide (1.25-7.5 mg/day) (in the concomitant sulfonylurea group, BMI 25.4 +/- 4.7, HbA1c 9.2 +/- 1.2%). BMI, HbA1c, serum lipid level, and body fat distribution, which were determined by computed tomography (CT) scan at the umbilical level, were measured and compared before and after troglitazone treatment. RESULTS: During the 6-month troglitazone treatment, HbA1c levels decreased and BMI increased in both groups. As for body fat distribution in the single-treatment group, visceral fat area (VFA) decreased (from 118.3 +/- 54.3 to 101.1 +/- 50.8 cm2; P < 0.001), and subcutaneous fat area (SFA) increased (from 189.7 +/- 93.3 to 221.6 +/- 101.6 cm2; P < 0.001), resulting in a decrease in visceral/subcutaneous (V/S) ratio (from 0.74 +/- 0.48 to 0.50 +/- 0.32; P < 0.001). In the concomitant sulfonylurea group, VFA was unchanged (from 108.1 +/- 53.5 to 112.5 +/- 59.9 cm2), while SFA increased (from 144.6 +/- 122.0 to 180.5 +/- 143.5 cm2; P < 0.01), thereby decreasing the V/S ratio (from 0.91 +/- 0.46 to 0.77 +/- 0.44; P < 0.01). The serum triglyceride level and the area under glucose curve during the 75-g oral glucose tolerance test decreased significantly in the single-treatment group. CONCLUSIONS: According to our data, troglitazone appears to promote fat accumulation in the subcutaneous adipose tissue rather than in the visceral adipose tissue in mildly obese Japanese people with type 2 diabetes. This shift of energy accumulation from the visceral to subcutaneous adipose tissue may greatly contribute to the troglitazone-mediated amelioration of insulin resistance. PMID- 10372241 TI - Konjac-mannan (glucomannan) improves glycemia and other associated risk factors for coronary heart disease in type 2 diabetes. A randomized controlled metabolic trial. AB - OBJECTIVE: To examine whether Konjac-mannan (KJM) fiber improves metabolic control as measured by glycemia, lipidemia, and blood pressure in high-risk type 2 diabetic patients. RESEARCH DESIGN AND METHODS: A total of 11 hyperlipidemic and hypertensive type 2 diabetic patients treated conventionally by a low-fat diet and drug therapy participated. After an 8-week baseline, all were randomly assigned to take either KJM fiber-enriched test biscuits (0.7 g/412 kJ [100 kcal] of glucomannan) or matched placebo wheat bran fiber biscuits during two 3-week treatment phases separated by a 2-week washout period. The diet in either case was metabolically controlled and conformed to National Cholesterol Education Program Step 2 guidelines, while medications were maintained constant. Efficacy measures included serum fructosamine, lipid profiles, apolipoproteins, blood pressure, body weight, and nutritional analysis. RESULTS: Compared with placebo, KJM significantly reduced the metabolic control primary end points: serum fructosamine (5.7%, P = 0.007, adjusted alpha = 0.0167), total:HDL cholesterol ratio (10%, P = 0.03, adjusted alpha = 0.05), and systolic blood pressure (sBP) (6.9%, P = 0.02, adjusted alpha = 0.025). Secondary end points, including body weight, total, LDL, and HDL cholesterol, triglycerides, apolipoproteins A-1, B, and their ratio, glucose, insulin, and diastolic blood pressure, were not significant after adjustment by the Bonferroni-Hochberg procedure. CONCLUSIONS: KJM fiber added to conventional treatment may ameliorate glycemic control, blood lipid profile, and sBP in high-risk diabetic individuals, possibly improving the effectiveness of conventional treatment in type 2 diabetes. PMID- 10372242 TI - Impaired glucose tolerance is a risk factor for cardiovascular disease, but not impaired fasting glucose. The Funagata Diabetes Study. AB - OBJECTIVE: To determine whether the new category of impaired fasting glucose (IFG) recently proposed by the Expert Committee of the American Diabetes Association is a risk factor for cardiovascular disease. RESEARCH DESIGN AND METHODS: Death certificates and residence transfer documents from the cohort population consisting of participants of the diabetes prevalence study in Funagata, Yamagata prefecture, Japan, 1990-1992, were analyzed up through the end of 1996. First, the cohort population was classified into three groups: normal glucose tolerance (NGT) (n = 2,016), impaired glucose tolerance (IGT) (n = 382), and diabetic (n = 253). Then the same population was reclassified into normal fasting glucose (NFG), IFG, and diabetic. The cumulative survival rates among the groups were compared using the classical life-table method, and age-adjusted analyses, the person-year method, and Cox's proportional hazard model were adopted. RESULTS: At the end of seven observed years, the cumulative survival rates from cardiovascular disease of IGT and diabetes were 0.962 and 0.954, respectively, both significantly lower than that of NGT (0.988). The Cox's proportional hazard model analysis showed that the hazard ratio of IGT to NGT on death from cardiovascular disease was 2.219 (95% CI 1.076-4.577). However, the cumulative survival rate of IFG from cardiovascular disease was 0.977, not significantly lower than that of NFG (0.985). The Cox's hazard ratio of IFG to NFG on death from cardiovascular disease was 1.136 (0.345-3.734), which was not significant either. CONCLUSIONS: IGT was a risk factor for cardiovascular disease, but IFG was not. PMID- 10372243 TI - Incidence of lactic acidosis in metformin users. AB - OBJECTIVE: The purpose of this study was to determine the incidence of lactic acidosis in a geographically defined population of metformin users. RESEARCH DESIGN AND METHODS: The study was based on a historical cohort from the Saskatchewan Health administrative databases. Individuals with a metformin prescription dispensed between 1980 and 1995 inclusive were eligible for the cohort. Person-years of exposure were calculated. Cases were defined by hospital discharge with a diagnosis of acidosis (International Classification of Diseases, Ninth Revision code: 276.2) and confirmation by chart review of a blood lactate level > or = 5 mmol/l. Death registrations of individuals dying within 120 days of a metformin prescription were also reviewed. RESULTS: During the study period, 11,797 residents received one or more metformin prescriptions, resulting in 22,296 person-years of exposure. There were 10 subjects who had hospital discharges with a diagnosis of acidosis. However, primary record review revealed only two cases with laboratory findings of elevated blood lactate levels, for an incidence rate of 9 cases per 100,000 person-years of metformin exposure. In both cases, other factors besides metformin could have contributed to the lactic acidosis. No additional cases were found on review of death registrations. CONCLUSIONS: From 1980 through 1995, the incidence rate of lactic acidosis was 9 per 100,000 person-years (95% CI 0-21) in patients dispensed metformin in Saskatchewan, Canada. This incidence rate was derived from a population with complete ascertainment of hospitalizations and deaths associated with lactic acidosis in metformin users. It is similar to previously published rates based on passive reporting of cases, and it is well below the lactic acidosis rate of 40 64 per 100,000 patient-years in patients prescribed phenformin. PMID- 10372244 TI - Perinatal and neonatal determinants of childhood type 1 diabetes. A case-control study in Yorkshire, U.K. AB - OBJECTIVE: To identify environmental factors that exert their effect in the perinatal and neonatal period and influence the subsequent onset of insulin dependent (type 1) diabetes during childhood. RESEARCH DESIGN AND METHODS: A population-based case-control study of data abstracted from the hospital obstetric and neonatal records of 196 children with type 1 diabetes and 325 age- and sex-matched control subjects. Analysis of matched sets by conditional logistic regression was conducted for a range of perinatal and neonatal factors. RESULTS: A significantly raised risk was observed for illnesses in the neonatal period (OR 1.61, 95% CI 1.06-2.44), the majority of which were infections and respiratory difficulties. Exclusive breast feeding as the initial feeding method was significantly protective (OR 0.65, 95% CI 0.45-0.94). There were no significant associations with high- or low-birth weight, being firstborn or small for-dates. All factors significant (5% level) for the entire dataset, that is, maternal age, type 1 diabetes in mothers, preeclampsia, delivery by cesarean section, neonatal illnesses, and initial breast feeding were modeled and the OR remained significant for all variables other than cesarean section. CONCLUSIONS: The findings are based on medical record data that cannot be subject to biased recall of mothers. Neonatal illnesses increased and initial breast feeding decreased the risk of childhood type 1 diabetes. Further determinants of risk are mothers with type 1 diabetes, older mothers, and preeclampsia during pregnancy. PMID- 10372245 TI - Implications of new diagnostic criteria for abnormal glucose homeostasis in women with previous gestational diabetes. AB - OBJECTIVE: To determine the consequences of applying revised American Diabetes Association (ADA) (1997) and World Health Organization (WHO) (1998) recommendations for the classification of glucose intolerance in women with previous gestational diabetes mellitus (GDM). RESEARCH DESIGN AND METHODS: There were 192 women with previous GDM who took an oral glucose tolerance test (OGTT) 1 86 months after delivery and were classified by WHO (1985), ADA (1997, fasting glucose), and revised WHO (1998) guidelines. RESULTS: Among the 165 women without a preexisting diagnosis of diabetes, WHO-1985 and ADA-1997 provided similar estimates of diabetes prevalence (13.3% vs. 11.5%) but widely differing estimates of impaired glucose homeostasis (31.5% impaired glucose tolerance [IGT] by WHO 1985 vs. 10.9% impaired fasting glucose by ADA-1997 criteria). Overall, 56 women (34%) showed a classification discrepancy between WHO-1985 and ADA-1997 criteria, including 44 with normal fasting glucose by ADA-1997 criteria, but abnormal 2-h glucose by WHO-1985 criteria (40 IGT, 4 diabetes). The cardiovascular risk profile of these women was more favorable than that of 18 women with impaired fasting glucose. WHO-1998 recommendations reproduced ADA-1997 findings when used as a fasting screen, but behaved similarly to WHO-1985 criteria when 2-h glucose values were also analyzed. CONCLUSIONS: All criteria produced similar estimates of diabetes prevalence. However, analyses based on a single fasting glucose screen (and a threshold of 6.1 mmol/l) failed to identify 60% of women with abnormal 2-h glucose levels. Screening women with previous GDM (and by analogy, other groups at high risk of diabetes) with a single fasting glucose has low sensitivity for the detection of abnormal glucose tolerance. Recent guidelines recommending this approach require reevaluation. PMID- 10372246 TI - Excess maternal transmission of type 2 diabetes. The Northern California Kaiser Permanente Diabetes Registry. AB - OBJECTIVE: To assess excess maternal transmission of type 2 diabetes in a multiethnic cohort. Previous studies have reported higher prevalence of diabetes among mothers of probands with type 2 diabetes than among fathers. This analysis is vulnerable to biases, and this pattern has not been observed in all populations or races. RESEARCH DESIGN AND METHODS: We assessed evidence for excess maternal transmission among 42,533 survey respondents with type 2 diabetes (probands) by calculating the prevalence of diabetes in their siblings and offspring. To assess data quality, we evaluated completeness of family history data provided. Accuracy of family information reported by probands was also evaluated by comparing survey responses in a subsample of 206 probands with family histories modified after further interviews with relatives. RESULTS: Siblings (n = 60,532) of probands with affected mothers had a greater prevalence of diabetes (20%) than those with affected fathers (17%) (P < 0.001 for adjusted odds ratios). Prevalence of diabetes was higher among the offspring (n = 72,087) of female (3.4%) versus male (2.2%) probands (P < 0.001 for adjusted odds ratios). These patterns were evident in all races and both sexes; however, the effect size was clinically insignificant in African-Americans and male offspring. In general, probands provided more complete data about diabetes status for the maternal arm of the pedigree than the paternal arm. Completeness of knowledge was not related to proband sex, but was related to education and race, and inversely to age. Accuracy of proband-reported family history was consistently good (kappa statistics generally > 0.70). CONCLUSIONS: Excess maternal transmission was observed in all races and both sexes, although the size of the excess was negligible in African-Americans and male offspring. Potential reporting and censoring biases are discussed. PMID- 10372247 TI - Birth weight, type 2 diabetes, and insulin resistance in Pima Indian children and young adults. AB - OBJECTIVE: To investigate the mechanisms underlying the association between birth weight and type 2 diabetes in a population-based study of 3,061 Pima Indians aged 5-29 years. RESEARCH DESIGN AND METHODS: Glucose and insulin concentrations were measured during a 75-g oral glucose tolerance test, and insulin resistance was estimated according to the homeostatic model (homeostasis model assessment insulin resistance [HOMA-IR]). Relationships between birth weight, height, weight, fasting and postload concentrations of glucose and insulin, and HOMA-IR were examined with multiple regression analyses. RESULTS: Birth weight was positively related to current weight and height (P < 0.0001, controlled for age and sex, in each age-group). The 2-h glucose concentrations showed a U-shaped relationship with birth weight in subjects > 10 years of age, and this relation was independent of current body size. In 2,272 nondiabetic subjects, after adjustment for weight and height, fasting and 2-h insulin concentrations and HOMA IR were negatively correlated with birth weight. CONCLUSIONS: Low-birth-weight Pimas are thinner at ages 5-29 years, yet they are more insulin resistant relative to their body size than those of normal birth weight. By contrast, those with high birth weight are more obese but less insulin resistant relative to their body size. The insulin resistance of low-birth-weight Pima Indians may explain their increased risk for type 2 diabetes. PMID- 10372248 TI - The 14-year incidence of lower-extremity amputations in a diabetic population. The Wisconsin Epidemiologic Study of Diabetic Retinopathy. AB - OBJECTIVE: To estimate the cumulative 14-year incidence of lower-extremity amputations (LEAs) and evaluate risk factors for LEA. RESEARCH DESIGN AND METHODS: Study subjects consisted of population-based cohorts of younger-onset (diagnosed before age 30 years and taking insulin, n = 906) and older-onset (diagnosed after age 30 years, n = 984) individuals with diabetes. Subjects participated in baseline (1980-1982), 4-year, 10-year, and 14-year examinations or interviews. LEAs were determined by history. RESULTS: The cumulative 14-year incidence of LEA was 7.2% in younger- and 9.9% in older-onset patients. In multivariable analyses based on the discrete linear logistic model, LEA in the younger-onset group was more likely for males (odds ratio [OR] 5.21 [95% CI 2.50 10.88]), older age (OR for 10 years 1.71 [1.30-2.24]), higher glycosylated hemoglobin (OR for 1% 1.39 [1.22-1.59]), higher diastolic blood pressure (OR for 10 mmHg 1.58 [1.20-2.07]), history of ulcers of the feet (3.19 [1.71-5.95]), and more severe retinopathy (OR for one step 1.16 [1.08-1.24]). In younger-onset patients aged > or = 18, pack-years smoked (OR for 10 years 1.20 [1.03-1.41]) was also associated with LEAs, and daily aspirin use was inversely associated (OR 0.11 [0.01-0.83]). In the older-onset group, LEA was more likely for men (2.66 [1.49, 4.76]) and if the subject had higher glycosylated hemoglobin (OR for 1% 1.25 [1.09-1.43]), higher pulse pressure (OR for 10 mmHg 1.19 [1.04-1.37]), history of ulcers (3.56 [1.84-6.89]), and more severe retinopathy (OR for one step 1.07 [1.00-1.13]). CONCLUSIONS: There are several risk factors for LEA with potential for modification and preventive strategies. PMID- 10372249 TI - A randomized double-blind trial of acarbose in type 2 diabetes shows improved glycemic control over 3 years (U.K. Prospective Diabetes Study 44) AB - OBJECTIVE: To determine the degree to which alpha-glucosidase inhibitors, with their unique mode of action primarily reducing postprandial hyperglycemia, offer an additional therapeutic approach in the long-term treatment of type 2 diabetes. RESEARCH DESIGN AND METHODS: We studied 1,946 patients (63% men) who were previously enrolled in the U.K. Prospective Diabetes Study (UKPDS). The patients were randomized to acarbose (n = 973), titrating to a maximum dose of 100 mg three times per day, or to matching placebo (n = 973). Mean +/- SD age was 59 +/- 9 years, body weight 84 +/- 17 kg, diabetes duration 7.6 +/- 2.9 years, median (interquartile range) HbA1c 7.9% (6.7-9.5), and fasting plasma glucose (FPG) 8.7 mmol/l (6.8-11.1). Fourteen percent of patients were treated with diet alone, 52% with monotherapy, and 34% with combined therapy. Patients were monitored in UKPDS clinics every 4 months for 3 years. The main outcome measures were HbA1c, FPG, body weight, compliance with study medication, incidence of side effects, and frequency of major clinical events. RESULTS: At 3 years, a lower proportion of patients were taking acarbose compared with placebo (39 vs. 58%, P < 0.0001), the main reasons for noncompliance being flatulence (30 vs. 12%, P < 0.0001) and diarrhea (16 vs. 8%, P < 0.05). Analysis by intention to treat showed that patients allocated to acarbose, compared with placebo, had 0.2% significantly lower median HbA1c at 3 years (P < 0.001). In patients remaining on their allocated therapy, the HbA1c difference at 3 years (309 acarbose, 470 placebo) was 0.5% lower median HbA1c (8.1 vs. 8.6%, P < 0.0001). Acarbose appeared to be equally efficacious when given in addition to diet alone; in addition to monotherapy with a sulfonylurea, metformin, or insulin; or in combination with more complex treatment regimens. No significant differences were seen in FPG, body weight, incidence of hypoglycemia, or frequency of major clinical events. CONCLUSIONS: Acarbose significantly improved glycemic control over 3 years in patients with established type 2 diabetes, irrespective of concomitant therapy for diabetes. Careful titration of acarbose is needed in view of the increased noncompliance rate seen secondary to the known side effects. PMID- 10372250 TI - Clinical correlates of plantar pressure among diabetic veterans. AB - OBJECTIVE: To assess the relationship between diabetes characteristics, medical history, foot deformity, sensory neuropathy, and plantar foot pressure. RESEARCH DESIGN AND METHODS: There were 517 subjects from a cohort of diabetic veterans enrolled in a prospective study of risk factors for foot complications who contributed 1,017 limbs for study. We interviewed subjects to collect data on demographics, diabetes characteristics, and medical history. A research nurse practitioner performed a directed physical exam of the lower extremities, assessing foot deformities and including quantitative sensory testing with a 5.07 monofilament. In-shoe foot-pressure measurements were obtained with F-scan insoles on subjects wearing their own footwear while walking 10 m at their usual pace. RESULTS: In univariate analyses, significant associations were seen between at least one measure of plantar pressure and body mass, sex, race, age, insulin use, certain foot deformities, plantar callus, and sensory neuropathy. Diabetes duration, HbA1c, and history of foot ulcer or amputation were unrelated to plantar pressure. In multiple regression analyses, body mass measured as log (weight), insulin use, white race, male sex, plantar callus, and diabetes duration were significantly related to certain pressures. Foot deformities were related primarily to forefoot pressures. With high pressure at two or more sites defined as the outcome, only body mass remained statistically significant as a predictor of this outcome in a backwards elimination logistic regression model. CONCLUSIONS: High in-shoe plantar pressure in diabetic subjects can be predicted in part from readily available clinical characteristics. The mechanisms by which these characteristics may be related to plantar pressure require further study. PMID- 10372252 TI - Effects of diabetes on cardiovascular drug metabolism. Emerging clinical implications. PMID- 10372253 TI - The European Association for the Study of Diabetes Annual Meeting, 1998. The U.K. Prospective Diabetes Study and other topics in type 2 diabetes. PMID- 10372251 TI - Impaired endothelium-dependent vasodilation in type 2 diabetes. Relation to LDL size, oxidized LDL, and antioxidants. AB - OBJECTIVE: To search for determinants of endothelial dysfunction in type 2 diabetes. RESEARCH DESIGN AND METHODS: We performed a comprehensive analysis of cardiovascular risk markers and measured blood flow responses to endothelium dependent (acetylcholine [ACh] and NG-monomethyl-L-arginine) and -independent (sodium nitroprusside [SNP]) vasoactive agents in 30 nonsmoking men with type 2 diabetes (age 51 +/- 1 years, BMI 27.8 +/- 0.4 kg/m2, HbA1c 7.4 +/- 0.3%) and 12 matched normal control men. RESULTS: ACh-induced vasodilation was 37% lower in type 2 diabetic (6.1 +/- 0.5) than in normal subjects (9.7 +/- 1.5 ml.dl-1.min-1, P < 0.01), while flows during SNP were similar (9.1 +/- 0.6 vs. 9.9 +/- 1.3 ml.dl 1.min-1, NS). The ratio of endothelium-dependent vs. -independent flow (ACh:SNP ratio) was 31% lower in type 2 diabetic (0.70 +/- 0.05) than in normal subjects (1.10 +/- 0.18, P < 0.01). Total (2.2 +/- 0.4 vs. 1.3 +/- 0.2 mmol/l, P < 0.05), VLDL, and intermediate-density lipoprotein triglycerides were significantly higher, and the mean LDL particle diameter was significantly smaller in type 2 diabetic than in normal subjects. The lag times for LDL oxidation by Cu2+ in vitro were similar in patients with type 2 diabetes (183 +/- 7) and in normal subjects (183 +/- 9 min, NS). Measured and calculated (sum of concentration of individual antioxidants in serum) total peroxyl radical-trapping capacities (TRAPs) were comparable between the groups. In the patients with type 2 diabetes, LDL size was significantly correlated with endothelium-dependent vasodilation (r = 0.43, P < 0.05), serum triglycerides (r = -0.75, P < 0.001), and the lag time for LDL oxidation in vitro (r = 0.38, P < 0.05). HbA1c was inversely correlated with the lag time for LDL oxidation in vitro (r = -0.41, P < 0.05) and TRAP. CONCLUSIONS: In summary, patients with type 2 diabetes exhibited impaired endothelium-dependent vasodilation in vivo, elevated serum triglycerides, decreased LDL size, and normal antioxidant capacity. Of these parameters, LDL size was significantly correlated with endothelial function. PMID- 10372254 TI - GAD antibodies in elderly men in different categories of glucose tolerance. PMID- 10372255 TI - Depression in subjects with type 2 diabetes. Predictive factors and relation to quality of life. PMID- 10372256 TI - Central pontine myelinolysis. An unusual complication of diabetes. PMID- 10372257 TI - Gastroparesis cured by gastrectomy. PMID- 10372258 TI - Implanted insulin pump may represent a chance for young women with unstable type 1 diabetes to give birth. PMID- 10372259 TI - Diabetic ketoacidosis with olanzapine treatment. PMID- 10372260 TI - Revised concept for the estimation of insulin sensitivity from a single sample. PMID- 10372261 TI - Acute hyperinsulinemia reduces plasma concentrations of homocysteine in healthy men. PMID- 10372262 TI - Switching insulin-sensitizing agents in patients with type 2 diabetes who require insulin. PMID- 10372263 TI - Antiproteinuric effect of pentoxifylline in patients with diabetic nephropathy. PMID- 10372264 TI - Potential impact of HbA1c determination on clinical decision making in patients with cystic fibrosis-related diabetes. PMID- 10372265 TI - Metformin and lactic acidosis. PMID- 10372266 TI - GAD antibodies in classification of Asian type 2 diabetes. PMID- 10372267 TI - Poor metabolic control decreases the growth velocity of diabetic children. PMID- 10372268 TI - Epidemiology and the natural course of inflammatory bowel disease. AB - Ulcerative colitis and Crohn's disease are inflammatory disorders of the gastrointestinal tract that are distributed unevenly within populations and throughout the world. Although the exact causes of inflammatory bowel disease (IBD) remain unknown, study of the epidemiology of IBD has provided insight into pathogenesis. This article examines the geographic, ethnic, and other trends of IBD; risk factors (including genetic and environmental); and the natural history of IBD. PMID- 10372269 TI - Current theories on the causes of inflammatory bowel disease. AB - The understanding of the pathogenesis of CD and UC has greatly expanded over the last decade. The combination of abnormalities in the immune system, the contribution of nonimmune cells in the intestinal mucosa, a variety of genetic risk determinants, and random environmental factors may all be necessary to induce what clinically presents as IBD. It is likely that several agents can initiate an immune response that in the intestinal microenvironment and the genetic background of the patient finally leads to pathology. PMID- 10372270 TI - Medical therapy for inflammatory bowel disease. AB - CD and UC represent a spectrum of chronic IBD that present in protean ways and are accompanied by a variety of systemic sequelae. Sulfasalazine and the newer 5 aminosalicylates are important in the management of mild-to-moderate disease, whereas corticosteroids remain the primary therapy for most patients with moderate-to-severe disease (Tables 2-5). The toxicities associated with long-term steroid therapy, combined with their ineffectiveness as maintenance medications, have led to increased use of immunomodulators, such as azathioprine and 6-MP, for the treatment of steroid-dependent and steroid-resistant IBD. Infliximab is a novel therapeutic adjunct for chronically active and fistulizing CD that will herald a new era of biologic therapy for IBD. Meanwhile, CSA remains an alternative to urgent colectomy in severe UC unresponsive to corticosteroids and also for CD patients with severe disease or refractory fistulas. Finally, continued insights into the etiopathogenic pathways in IBD will provide evolving and innovative approaches until the eventual causes and cures are elucidated. In the meantime, clinicians should remain optimistic regarding current ability to reduce the morbidity and maintain the quality of life for patients suffering with these frustrating diseases. PMID- 10372271 TI - Novel therapies for inflammatory bowel disease. AB - Looking back at successes and failures in newer approaches to treating IBD, it is tempting--although still difficult--to draw conclusions about pathogenesis. When a therapy proves effective, do clinicians truly know how it works? Even with a therapy as specific as anti-TNF antibody, it is not clear if the benefit is attributable to simple binding and clearance of TNF-alpha or to binding on the cell surface and subsequent deletion of the activated macrophage. When a drug appears to be less effective than preclinical models suggest, can failures in effectiveness from delivery or dosing be differentiated? The disappointing results of clinical trials with IL-10--so at odds with the prediction of benefit from animal models--bring into question the validity of those models as well as the soundness of design of the clinical trials on which efficacy of IL-10 is judged. The variability of response even to the most narrowly targeted agents suggests that these diseases are far more heterogeneous in humans than in their murine counterparts. Clinicians are only just beginning to recognize subclinical markers of response, and it may soon be possible to predict response on the basis of genetic composition. For the moment, however, the field of pharmacogenetics is embryonic. Challenges in developing new therapeutic strategies include not only identifying novel agents, but also improving the definitions of clinical endpoints and defining efficacy at the biologic level. Only through considered evaluation of clinical evidence may clinicians determine which therapies should remain novelties and which should become an accepted part of the armamentarium. PMID- 10372272 TI - Medical therapy of specific clinical presentations. AB - A pluridisciplinary approach that integrates medical therapy with surgery and other aspects of patient care, such as nutritional and psychosocial support, is essential to the management of patients with inflammatory bowel disease (IBD). Despite new medical therapies, such as 5-amino-salicylic acid compounds, steroids, and immunomodulators, the treatment of patients with IBD remains challenging. Success depends on the appropriate use of the available medications in relation to the severity and localization of the disease. The introduction of novel immunomodulating agents such as antitumor necrosis factor alpha is likely to have a major influence on the current therapeutic strategies. This article describes the use of the available medications in the most common clinical presentations of IBD. PMID- 10372273 TI - Surgical therapy for ulcerative colitis and Crohn's disease. AB - Over the past 2 decades there has been considerable progress in the surgical management of inflammatory bowel disease. Crohn's disease is a chronic, nonspecific inflammatory disease of the gastrointestinal tract of unknown cause. It involves mainly the ileum, colon, and rectum, most often producing symptoms of obstruction or localized perforation with fistula. Although surgical treatment is palliative, operative excision in combination with strictureplasty, where appropriate, provides effective symptomatic relief and reasonable long-term benefit. Chronic ulcerative colitis is a diffuse inflammatory disease of the mucosal lining of the colon and rectum. Total removal of the colon and rectum provides a complete cure. Newer surgical alternatives, developed over the last 2 decades, have eliminated the need for a permanent ileostomy following definitive resection of the involved colon and rectum. PMID- 10372274 TI - Imaging modalities in inflammatory bowel disease. AB - This article provides an in-depth review of the radiologic and endoscopic imaging techniques used in the evaluation and management of inflammatory bowel disease (IBD). The use of imaging studies to diagnose IBD and to differentiate ulcerative colitis from Crohn's disease is discussed. The evaluation of suspected complications associated with IBD, including strictures and fistulous disease, as well as surveillance for colorectal cancer are also addressed. PMID- 10372275 TI - Nutrition and inflammatory bowel disease. AB - This article reviews the nutritional aspects of inflammatory bowel disease (IBD) including the mechanisms and manifestations of malnutrition and the efficacy of nutritional therapies. Nutrient deficiencies in patients with IBD occur via several mechanisms and may complicate the course of the disease. Nutritional status is assessed by clinical examination and the use of nutritional indices such as the Subjective Global Assessment of nutritional status. Nutritional intervention may improve outcome in certain individuals; however, because of the costs and complications of such therapy, careful selection is warranted, especially in patients presumed to need parenteral nutrition. PMID- 10372276 TI - Inflammatory bowel disease in pediatric and adolescent patients. AB - IBD is a chronic pediatric disease that needs to be treated by a team of experts consisting of pediatricians, pediatric gastroenterologists, psychologists, nutritionists, social workers, and nurses. A critical factor in successful management of this disease is the willingness of the patient to participate and cooperate with the team. Parents and patients must be educated and supported to treat these disorders effectively. Much further research is necessary to understand the specific causative and therapeutic issues unique to young patients with IBD. PMID- 10372277 TI - Cancer risk in patients with inflammatory bowel disease. AB - Patients with inflammatory bowel disease (IBD) are at increased risk for developing cancer of the gastrointestinal tract, particularly colorectal cancer. Because of the relative rarity of IBD in the general population, it has been difficult to quantify this risk. Efforts to reduce the risk have included both prophylactic surgery and endoscopic screening programs. Because of the potential impact on quality of life and life expectancy, the optimal strategy for reducing this risk has not been defined. This article reviews the current literature relating to the risk of cancer for patients with IBD and methods to reduce this risk. PMID- 10372278 TI - The atypical colitides. AB - Collangenous colitis is a clinicopathologic syndrome characterized by (1) chronic watery diarrhea and crampy abdominal pain and (2) distinctive colorectal histopathology that includes a subepithelial collagen band, prominent chronic inflammation in the lamina propria, and increased intraepithelial lymphocytes. Lymphocytic colitis has similar clinical features to collangenous colitis, the main symptom being chronic watery diarrhea. Diversion colitis is an inflammatory process that arises in segments of the large intestine that are excluded from the fecal stream. This condition usually occurs in patients with ileostomy or colostomy when a mucous fistula or Hartmann's pouch has been left. PMID- 10372279 TI - Hepatobiliary manifestations of inflammatory bowel disease. AB - PSC is the most common of the clinically significant hepatobiliary diseases seen in association with IBD, with an incidence that varies from 2.5% to 7.5%. Conversely, 50% to 75% of patients with PSC have IBD. This high degree of association suggests a common pathogenetic mechanism; however, no causal relationship has been established. The etiopathogenesis of PSC remains poorly understood, despite a large number of studies looking at differing hypotheses. The diagnosis is usually established by cholangiography. Liver biopsy can sometimes be helpful in diagnosing pericholangitis. There is a significant overlap of the histology with chronic hepatitis. Serum markers have been studied for diagnosing PSC, particularly for early diagnosis of cholangiocarcinoma, but none have shown the high sensitivity and specificity needed to use them clinically. PSC usually progresses insidiously and eventually leads to cirrhosis. Despite progress in early recognition, optimal management of patients with PSC remains a challenge requiring a multidisciplinary approach among hepatologists, endoscopists, surgeons, and interventional radiologists. Colectomy for ulcerative colitis does not alter the natural history of PSC. There is a high (10% to 15%) incidence of cholangiocarcinoma in patients with PSC. This incidence along with the risk of colon cancer in patients with ulcerative colitis makes it necessary to follow these patients closely. A number of pharmacologic therapies have been evaluated, but none has proven successful in slowing the progression of PSC or prolonging survival. Endoscopic therapy has a proven utility in treating complications of recurrent cholangitis or worsening jaundice in the setting of a dominant stricture, but endoscopy has not been shown to improve survival or decrease the need for liver transplantation. Liver transplantation is life-saving for patients with advanced PSC. Pericholangitis, gallstones, and chronic hepatitis are additional disorders noted in association with IBD, but they are much less common and easier to manage than PSC. PMID- 10372280 TI - Depressive symptoms in the perimenopause: prevalence, assessment, and guidelines for treatment. AB - This review describes the biological changes occurring in perimenopause and analyzes epidemiological studies that shed light on the relationship between perimenopause and mood. The role of estrogen as a treatment for depressive symptoms is also examined. We found that a positive association may exist between depressive symptoms and the perimenopause, and that a prior history of depression may be associated with such symptoms. In most of the studies reviewed, the use of estrogen in replacement doses appears to improve depressive symptoms in perimenopausal patients who do not have major depression. We suggest an approach to the treatment of middle-aged women presenting with such symptoms. No careful study of the incidence of DSM-IV major depression associated with perimenopause has been done, and the efficacy of estrogen as a primary or adjunctive treatment for the disorder during perimenopause is unclear. PMID- 10372281 TI - The effect of comorbid substance use disorders on the course of bipolar disorder: a review. AB - Population-based studies have documented that among all patients with major psychiatric disorders, those with bipolar disorder have the highest prevalence of comorbid substance abuse and dependence. The cause of this high comorbidity rate has not been clearly established, and the relationship is probably bidirectional. Articles published in English from 1980 through 1997 containing the terms comorbidity, mania, outcome, and substance use were identified by searching Medline and then the bibliographies of the articles identified in this search. The literature review showed several risk factors to be associated with comorbid substance use disorders in bipolar disorder patients: early age of onset, gender, family history of substance use disorders, and presence of mixed mania. Methodological enhancements that have helped to advance understanding in this area include distinguishing between primary and secondary disorders, between the different subtypes of bipolar disorder, and between first and subsequent episodes of illness. In order to determine the temporal sequence of onset, longitudinal studies initiated at the onset of either illness need to be pursued. Increased understanding of the association between bipolar disorder and comorbid substance use disorder will facilitate improved detection and intervention, as well as more effective preventive measures that could improve outcome for patients with bipolar disorder. PMID- 10372282 TI - Anxious traits, anxious attachment, and anxious-cluster personality disorders. AB - This paper proposes a predisposition-stress model for the pathogenesis of anxious cluster personality disorders. In this model, genetically influenced anxious traits predispose individuals to such disorders, while psychological factors promoting anxious attachment, as well as social factors leading to discrepancies between traits and social expectations, constitute stressors. Three hypotheses are derived from this model: (1) anxious traits are necessary conditions for the anxious cluster, but disorders develop as a result of cumulative adversities; (2) gene-environment interactions increase the likelihood of adversities associated with these disorders; and (3) social context has an important effect on the pathogenesis of this group of disorders. This theoretical model could have clinical implications for the management of a problematic group of patients. PMID- 10372283 TI - Irritable bowel/irritable mood: the mind/gut connection. PMID- 10372284 TI - History of managed behavioral health care and speculations about its future. PMID- 10372285 TI - Psychoanalytic and biological perspectives on lesbian patients: why developmental themes are more important in psychotherapy. PMID- 10372286 TI - Screening for psychiatric disorders in primary care settings. PMID- 10372287 TI - Identifying critical periods of neurodevelopmental risk: the turning point in schizophrenia research. PMID- 10372289 TI - Choices in antipsychotic therapy in schizophrenia. AB - Pharmacotherapy for the treatment of schizophrenia now consists, for the most part, of two groups of agents. The conventional antipsychotic agents are exemplified by chlorpromazine and haloperidol, and the atypical agents by clozapine, risperidone, olanzapine, and quetiapine. In this article, the history of the development of these two groups, and their advantages and disadvantages, are reviewed. Effectiveness, side-effect burden, mode of delivery, and cost are discussed. The new practice of "stalled or reversed taper" is described. The clinician now has a wider range of options from which to choose, but many clinical questions remain unanswered. PMID- 10372288 TI - Schizophrenia: improving outcome. AB - Therapeutic advances over the last four decades have enabled most persons with schizophrenia to live in the community. Nevertheless, the majority will continue to experience various symptoms and to have social and cognitive disabilities. With the development of new medications and psychosocial interventions, outpatient status can no longer be viewed as a satisfactory final outcome. This article presents the current state of schizophrenia therapeutics in a variety of clinically relevant situations: first-episode psychosis, treatment-resistant psychosis, chronic, relapsing psychosis, continuous poor functioning, and chronic psychosis not responsive to pharmacotherapy. The first-line atypical antipsychotics should generally be used, mainly because of their comparatively benign side-effect profiles, and they should be given as early as possible in the illness. The clinician should not be quick to accept persistent psychosis; the second-line atypical clozapine should be tried early in the course of the disease in patients showing treatment resistance. For patients residing with their families, educational and supportive family interventions have an important effect on relapse prevention; for those who live on their own and suffer frequent relapses, Assertive Community Treatment may be helpful. Patients with psychosis that is not responsive to pharmacotherapy may benefit from specific modalities of cognitive-behavioral therapy currently being developed, while persons with persistent negative symptoms and limited social competence may find social-skills training useful. In addition, new programs of supported employment may enable some patients to maintain competitive employment. PMID- 10372290 TI - Developmental psychiatry: is there any other kind? AB - This paper describes the importance of the developmental perspective in psychiatry and addresses the lack of a developmental focus in the DSM-based descriptive empirical model. Although the publication of DSM-III in 1980 represented a "breakthrough" in psychiatry, the revisions of its diagnostic framework over the subsequent two decades have not adapted to the rapidly evolving changes in the field. In this paper we argue that, like once-grand theories, the breakthroughs in the diagnostic framework need to transform. The developmental perspective provides an interdisciplinary and conceptual framework linking facts and theories. It is inherent in different aspects of psychiatry and readily accommodates the descriptive-empirical model by means of inclusive concepts borrowed from developmental psychopathology and psychobiology. It also makes important contributions to a process-oriented approach to measurement. Developmentally operationalized and multidimensional constructs stand to broaden psychiatric domains beyond diagnosable disorders. This argues for preventive and early treatment interventions for a variety of mild, subthreshold, or delayed symptoms of various conditions, based on an understanding of the causal mechanisms and developmental processes involved. PMID- 10372291 TI - Events in the life of the therapist: the effect on transference and countertransference. AB - Interactions between therapist and patient are influenced by the feelings and impressions that develop during the therapeutic relationship. The terms transference and countertransference are used to describe this process. However, attention must also be paid to the possible effects that events in the personal life of the therapist can have on therapy. The therapeutic interchange can be disrupted when this factor is ignored. PMID- 10372292 TI - The threat of the housing inspector: a case of hoarding. PMID- 10372293 TI - The use of natural remedies in psychiatry: a commentary. PMID- 10372294 TI - [Current aspects on differentiating thoracic pain symptoms]. PMID- 10372295 TI - Abnormal cardiac nerve function in syndrome X. AB - Syndrome X is likely to be caused by a dysfunction of small coronary arteries. Several authors suggested that an increased adrenergic activity could be involved in the pathogenesis of syndrome X, but studies investigating this topic by indirect methods led to conflicting results. We directly investigated cardiac sympathetic nerve function in syndrome X by myocardial radionuclide studies with 123I-metaiodobenzylguanidine (MIBG). Twelve syndrome X patients and 10 healthy controls were enrolled in the study. Cardiac MIBG uptake was assessed calculating the heart/mediastinum (H/M) ratio and a semiquantitative MIBG uptake score. Cardiac MIBG images were normal in all but 1 of controls (10%). Conversely, abnormalities in cardiac MIBG uptake were found in 9 syndrome X patients (75%, p < 0.01). In 5 patients the heart was totally or almost totally invisible on radionuclide MIBG images, while regional defects were found in other 4 patients. The H/M ratio was lower and cardiac MIBG uptake score strikingly higher in syndrome X patients. At 3 hours the H/M ratio was 1.70 +/- 0.6 in patients and 2.19 +/- 0.3 in controls (p = 0.03), while MIBG uptake score was 36.7 +/- 31 and 4.0 +/- 2.5 (p = 0.003) in the 4 groups, respectively. There were no differences between patients and controls in lung and salivary MIBG uptake. Reversible perfusion defects on stress thallium scintigraphy were found in 5 syndrome X patients (45%), all of whom also had abnormal MIBG scintigrams, while all 3 patients with normal MIBG scintigraphy also had normal thallium images. Thus, the function of efferent cardiac adrenergic nerve fibers is strongly impaired in the majority (i.e., 75%) of syndrome X patients. This abnormal function likely contributes significantly to the pathophysiologic and clinical features of syndrome X. We speculate that also the increased perception of cardiac pain reported in these patients could be an expression of the abnormal function of cardiac nerves, reflecting alterations of afferent nociceptive cardiac nerve fibers, as the abnormalities in MIBG uptake reflect alterations of efferent cardiac adrenergic nerve fibers. PMID- 10372296 TI - [Diagnosis of functional heart complaints from the psychosomatic viewpoint]. AB - Disorders of the cardiovascular system are common. Heart pain is one of the most frequent complaints leading patients to seek medical help. Although psychologically conspicuous behaviour in patients with functional cardiac complaints are well known, they are--if at all--diagnosed quite late. Descriptive diagnostics of functional cardiac complaints according to the International Classification of Diseases (ICD-10, Chapter 5) are discussed (Figure 1). Possible physical causes of the disease must first be excluded. In a second step it must be clarified whether the complaints even those non-verbally conveyed are due to psychic illness in a narrower sense. Anxiety and depressive disorders must be taken into consideration here. If the patient demonstrates an avoidance behavior in the case of anxiety, than an agoraphobia can be assumed; in episodic paroxysmal fear on can assume panic attacks in which vegetative anxiety equivalents such as shortness of breath, numbness, palpitation of the heart, tachycardia and chest pain are prominent often accompanied by trembling, perspiration, nausea and dizziness. The different depressive disorders are characterized by a dejected mood, loss of interest, loss of enthusiasm and drive reduction; the disorders are divided up according intensity and course. Within the scope of depressive physical symptoms, frequently unpleasant sensations and pain in the chest area are described along with concern, despair, and an increase in self-observation. If no psychic disturbance in a narrower sense can be diagnosed, then the diagnosis of a somatoform disorder allows for this behavior. It is characteristic for this category of illness that the repeated presentation of physical symptoms in connection with the persistent demand for medical treatment may be observed although no physical causes can be demonstrated. The patients insist that their complaints are of a physical origin despite the doctor's assertion that this is not the case. If the symptoms are related to vegetative innervated organs then one speaks of somatoform autonomous functional disorders (F45.3, Table 1). Cardiovascular disorders fall within this scope. Further diagnoses within the spectrum of somatoform disorders are hypochondric and somatization disorders which demonstrate a variety of symptoms and inconsistent and frequently changing complaints. If a descriptive diagnosis can correspondingly be made then further analysis of the disorder must be carried out in order to reach an indication for psychotherapeutic treatment. From a psychodynamic point of view, the personality- and conflict-related background of the disturbance is relevant. Quite often unconscious ambivalent separation conflicted--be they real are fantasized situations of being left or being left alone--may be observed to trigger cardiovascular symptoms. In the cognitive behavioral therapeutic tradition an exact analysis of the patients symptomatology is carried out in which prior and actual cause factors of the symptoms are looked for. Irrespective of the different approaches, information on the context of the complaints both on a biological, intrapsychic and interpersonal level is necessary for psychosomatic diagnostics. The better the causal conditions are known on the basis of which functional cardiovascular complaints have arisen, the easier it is to recognize those factors that will influence a change and allow a therapeutic approach. This is best done in cooperation with practitioners and internists who still have a key position in the diagnosis and treatment of patients with functional cardiac disorders. The ways and means in which they conduct the anamnesis is decisive in leading their patients to regard psychosomatic diagnostics as being either stuck in the so-called "psycho corner" or as a helpful relationship which they can accept. PMID- 10372298 TI - Chest pain after coronary interventional procedures. Incidence and pathophysiology. AB - Chest pain following successful percutaneous coronary interventions is a common problem. Although the development of chest pain after coronary interventions may be of benign character, it is disturbing to patients, relatives and hospital staff. Such pain may be indicative of acute coronary artery closure, coronary artery spasm or myocardial infarction, but may also simply reflect local coronary artery trauma. The distinction between these causes of chest pain is crucial in selecting optimal care. Management of these patients may involve repeat coronary angiography and additional intervention. Commonly, repeat coronary angiography following percutaneous transluminal coronary angioplasty (PTCA) in patients with chest pain demonstrates widely patent lesion sites suggesting that the pain was due to coronary artery spasm, coronary arterial wall stretching or was of non cardiac origin. As reported by the National Heart, Lung and Blood Institute PTCA Registry, 4.6% of patients after angioplasty have coronary occlusions, 4.8% suffer a myocardial infarction, and 4.2% have coronary spasm. The frequency of chest pain after new device coronary interventions (atherectomy and stenting) seems to be even higher. However, only the minority of patients with post procedural chest pain have indeed an ischemic event. Therefore, the vast majority of patients have recurrent chest pain without any signs of ischemia. There is some evidence that non-ischemic chest pain after coronary interventions is more common after stent implantation as compared to PTCA (41% vs. 12%). This may be due to the continuous stretching of the arterial wall by the stent as the elastic recoil occurring after PTCA is minimized. In conclusion, chest pain after coronary interventional procedures may potentially be hazardous when due to myocardial ischemia. However, especially after coronary stent placement, cardiologists must consider "stretch pain" due to the overdilation and stretching of the artery caused by the stent in the differential diagnosis. Clinically, it is, therefore, important to recognize that in addition to ischemia-related chest pain other types of chest pain do exist with cardiac origin. PMID- 10372297 TI - Visceral chest pain in unstable angina pectoris and effects of transcutaneous electrical nerve stimulation. (TENS). A review. AB - A substantial proportion of patients with chest pain referred to hospital, show signs of coronary artery disease. Anginal pain could be conceptualized as a warning signal for coronary artery disease and impending death. But, for many reasons this theory is partly disputed. Firstly, not all ischemic episodes are accompanied by anginal pain (silent ischemia). Secondly, chest pain indistinguishable from true angina pectoris may be the result of other abnormalities of thoracic viscera. Nevertheless acute severe cardiac ischemia often gives rise to anginal chest pain. Unstable angina pectoris is carrying a higher risk for future events in spite of intensive medical treatment. A special problem are patients awaiting coronary intervention because of severe ischemia and maximum medical treatment, who experience ischemic pain. New treatment regimens are needed for these patients. This review discusses the symptom of visceral pain from the heart, angina pectoris, its relation to ischemia and unstable angina pectoris. It also addresses the role of afferent nerve stimulation (transcutaneous electrical nerve stimulation, TENS) in the treatment of severe angina pectoris as well as recent findings of TENS applicability in unstable angina. PMID- 10372299 TI - [Angina pectoris in extracoronary diseases]. AB - Chest pain can arise from cardiovascular or noncardiovascular causes. Among the latter are the skin, the chest wall, intrathoracic structures, or subdiaphragmatic organs. The problem to attribute the chest discomfort to either the heart or extracardiac organs arises because the heart, pleura, aorta, and esophagus are all supplied by sensory fibers from the same spinal segments. In contrast to the diseases mentioned above, angina pectoris in sensu strictu is defined as chest pain or discomfort of cardiac origin that arises because of temporary imbalance between myocardial oxygen supply and demand. The metabolic oxygen requirements of the myocardium are essentially dictated by myocardial contraction since only a fraction of the consumed oxygen is needed by the quiescent heart. Therefore, the factors that primarily influence myocardial oxygen consumption include heart rate, the force of cardiac contraction, and myocardial wall tension, as determined by pressure (afterload), volume (preload), and wall thickness. Extracoronary diseases, e.g. hypertensive heart disease, aortic stenosis or cardiomyopathies, can influence these factors and induce angina pectoris (Figure 1). On the other hand, different diseases influencing the oxygen supply, e.g. anemia, can cause angina pectoris, too. In addition, the modulation of the coronary tone by mediators and cytokines can cause angina, coronary spasm being one example. The neurophysiological substrate of angina pectoris are ganglia which are present within the heart, particularly in epicardial fat. The sympathetic nervous system is the main conveyer of afferent pain fibers from the heart and pericardium, but many fibers may travel by the vagus and the phrenic nerves. Therefore, multiple thoracic structures may cause similar pain syndromes in the distressed patient. The blood supply of intrinsic cardiac ganglia arises primarily from branches of the proximal coronary arteries. Adenosine, among a number of substances, can modulate the activity generated by cardiac afferent nerve endings and intrinsic cardiac neurones. During myocardial ischemia adenosine is released in large quantities into the interstitial space. Given as an intravenous bolus to healthy volunteers or to patients with ischemic heart disease and angina pectoris, adenosine provokes angina pectoris-like pain, which is similar to habitual angina pectoris with regard to quality and location. But other mediators (e.g. bradykinin, histamine, prostaglandins, potassium, lactate) can be involved in the development of angina pectoris, too. As most emphasis should be given to the most serious causes first, the cardiologist has to consider ischemic cardiac disease in the differential diagnosis of nearly every case of acute chest pain. The differential diagnosis contains several causes of nonischemic cardiac chest pain. Dissecting aortic aneurysm may cause severe anterior chest pain that can be mistaken for myocardial infarction. Patients frequently will note the sudden onset of the pain rather than the relatively slower onset of ischemic pain. Furthermore, they feel as a tear and describe it as the most severe pain they have ever had. Pericarditis can be characterized as a sharp precordial knife-like pain that is often increased by lying down, breathing, swallowing, or any other thoracic motion. Radiation of pericardial pain is often relieved by sitting up or leaning forward. It may involve the shoulders, upper back, and neck because of the irritation of the diaphragmatic pleura. Acute pulmonary embolism is associated with severe chest pain. It may mimic acute myocardial infarction. Pulmonary embolism should be suspected when dyspnea or tachypnea seems to be disproportionate to the severity of the chest pain. Diffuse esophageal spasm is the extracardiac condition that is confused most often with ischemic cardiac chest pain. This pain presents as a deep thoracic pain that may be present over most of the thorax. It may extend down the anterome PMID- 10372300 TI - Aortic pain: the renaissance of cardiovascular pain and the detection of aortopathy. AB - Our 19th century predecessors considered the aorta as a source of cardiovascular pain associated with inflammatory aortitis, arterial hypertension, aortic aneurysms, aortic dissection, and aortic valve disease. However, during the 20th century epidemic of coronary artery disease clinicians became concerned with the syndromes associated with myocardial ischemia and infarction, relegating aortic pain syndromes to the role of a differential, "rule out", or diagnosis of exclusion rather than a primary diagnosis. We re-focus attention on a more global approach to cardiovascular pain, approaching thoracic aortic pain syndromes as primary diagnoses, while considering the dynamics and various stages of development of aortic pain syndromes, set within the clinical environment in which these syndromes arise. The central role of aortopathy is our underlying theme since the detection and clinical recognition of aortopathic disorders provide the template for identification of the population at risk for aortic pain syndromes. Clinical history, pedigree development, phenotype recognition, analysis of the elastic properties of the aorta, use of the wide range of sophisticated imaging techniques, and phenotype-genotype correlations provide the bases for the recognition, diagnosis, and management of aortopathy within the clinical setting. A futuristic anticipatory approach towards the diagnosis of aortopathy is outlined with emphasis on earlier recognition and informed clinical management ultimately leading to prevention of the acute and dynamic aortic complications. PMID- 10372301 TI - [Significance of esophagocardiac reflexes for inducing thoracic pain]. AB - In the clinical setting the cardiologists' interest is focussed on the esophagus as a potential source of thoracic pain as a differential diagnosis of angina pectoris. However, visceral afferences originating in the mucosal wall of the esophagus activated by acid exposure may also influence cardiac function. The available data convincingly demonstrate a reduction of the exertional angina threshold and changes of the ECG (ST segment depression and arrhythmia). These effects are most likely due to a reduced coronary blood flow. PMID- 10372303 TI - [Experiences with ambulatory cardiologic phase II rehabilitation]. AB - The phase II cardiac rehabilitation in Germany differs markedly from other European countries and the USA. Most of the patients enter a 3-week full residential program. In contrast we developed an outpatient phase II cardiac rehabilitation program. Since 1979 we treated more than 8,500 patients with different indications (i.e. after myocardial infarction, coronary bypass surgery, valve replacement and reconstruction). Patients with a daily commuting time over 60 minutes are not suitable for outpatient rehabilitation. Our model corresponds to the German intrahospital rehabilitation. The rehabilitation is carried out in 3 weeks offering approximately 66 hours of therapy. Groups of 8 patients with a similar level of physical capacity stay together during the rehabilitation. A comprehensive program with exercise training, physical therapy, psychological support, education in life style changes and risk factor modification has been developed. The compliance of the patients as well as the acceptance by the family are excellent. Long-lasting reduction in LDL cholesterol levels and increments in work-load capacities have been demonstrated. A high percentage of patients returned to work. Cost analysis demonstrates a reduction up to 40% in comparison to the full residential program. Therefore the outpatient phase II cardiac rehabilitation is a good alternative especially in urban areas. PMID- 10372302 TI - [Arterial myocardial revascularization in the 9th decade of life. Personal results and review of the literature]. AB - The rate of the population being 80 years of age and even older, has an increasing tendency in the Federal Republic of Germany. In 1996, a total of 87,372 patients received surgery supported by the heart-lung-machine, 2,383 patients out of these (2.7%) were 80 years of age and older. In view of the limited life expectance, the arterial revascularization in this age category is faced with controverse discussions. We analysed our patients in relation to this aspect. Between January 1, 1995 and June 30, 1997, 4,338 patients underwent surgery supported by the heart-lung-machine. Hundred and fifty-five out of these (3.6%) were in the 9th decade of life. Seventy-seven patients out of the 155 (49.7%, 34 women, 43 men, 80 to 88 years old, mean: 82 years of age) underwent an isolated myocardial revascularization. We performed 55 (71%) elective, 16 (21%) urgent and 6 (8%) emergency surgeries. Twelve patients (15.6%) solely received venous bypasses (Group I), 65 (84.4%) additionally also received unilateral bypasses of the internal mammaria artery (IMA) (Group II). Three patients died at our facility (3.9%), 3 further patients died during the follow-up treatment in outlying hospitals, the in-patient mortality rate in Group I therefore presented a rate of 8.3%, in Group II 7.7% and in total, a rate of 7.8%. In 1996, the in patient mortality rate could be reduced to 3.6%. The follow-up observation time ranged between 7 and 138 weeks (median 44 weeks). The survival rate for patients with an IMA-bypass after 1 year was 86.3%, after 2 years 77%, and for the entire collective 85.3% and 75%. Whereas 96% of the patients could pre-operatively be related to Class III or IV of the NYHA-classification, 55 of the 63 survivors (87%) belonged to Class I (6%) or II (81%). Two Group I patients (22.2%), 3 Group II patients (5.6%) and 7.9% of the total collective complained about repeated angina symptoms. The myocardial revascularization with the internal mammaria artery performed on patients in the 9th decade of life, achieves an acceptable morbidity and mortality compared to solely venous coronary bypasses. The more prolonged follow-up observation period will clarify, whether the arterial myocardial revascularization also proves to be the superior method in this age category. PMID- 10372305 TI - [Ambulatory phase II rehabilitation of heart patients in the Rhine-Main district hospital: the Frankfurt model]. AB - In the early 90s the German approach to phase II cardiac rehabilitation differed markedly from that which was usually practiced in the anglosaxon countries, as it covered almost exclusively a 4-week comprehensive, residential program far away from home in an often idyllic spa. However as many as approximately 60% of the cardiac patients did not take advantage of this offer, mainly because they felt separated from their families in residential programs. Therefore, because of good experiences with outpatient programs in other countries and in order to minimize costs, the so-called Frankfurt model, an outpatient phase II rehabilitation program, was performed at the University hospital in Frankfurt. The aim of this innovative program was to find out, whether it was as effective as the German "gold standard" of residential rehabilitation and whether is was practicable at a hospital. The program was carried out in a pilot phase over 1 year according to the standards of comprehensive cardiac rehabilitation in 41 patients, who had a follow-up examination 6 months later. The outcomes were compared with results of residential programs, which, however, were not collected in a controlled way. The short- and medium-term results concerning somatic outcomes, e.g. the risk factor profile or the improvement of the maximum work capacity, were equal to the residential programs. The percentage and timing of return-to-work, however, was much higher in the outpatient program obviously due to selection phenomena. The rehabilitation program at the hospital was practicable and safe. Under given conditions and meeting the standards for comprehensive cardiac rehabilitation an outpatient phase II rehabilitation program is practicable and safe with good short- and medium-term results also at a hospital. Such a program offers a good alternative to patients who do not want to rehabilitate under strictly residential conditions far away from home. PMID- 10372306 TI - [Partial inpatient cardiologic after-care]. AB - In Germany the in-hospital rehabilitation of patients with cardiac diseases is standard. Especially because of criticism of effectiveness and wasting of resources we evaluated in a pilot study a rehabilitation program with an in hospital start, followed facultatively on ambulatory basis at the same institution, at the cardiac rehabilitation clinic. The aims of the study were to find out the acceptance of the the program by the patients, the safety and the impact of the facultative ambulatory rehabilitation program on cardiovascular risk factors. From January 1993 to December 1995 the primary enclosing criteria (cardiac disease suitable for rehabilitation, age < 70 years, lodging < 40 km) were fulfilled by 612 patients, 122 female. 268 (43.8%) were disclosed on medical grounds because of the rigid pilot character of the study, for 74 (12.1%) patients was the participation on organisational grounds not possible. Sixty-six of the remaining 270 patients preferred the facultative continuation of the rehabilitation on ambulatory basis for 1 or 2 weeks, the entire rehabilitation lasting up to 4 weeks. During the ambulatory phase the patients slept at home, otherwise these patients followed the same procedure as the in-hospital participants. Despite of severe medical concerns 4 further patients wished to participate on facultative ambulatory continuation of the rehabilitation, among these were 2 women, only 5 of the total 70 patients of the facultative ambulatory program group being female. There were no significant differences among 24 patients with CAD of both groups during the rehabilitation and in a control after 14.8 months (Table 3). The impact on serum total cholesterol and LDL cholesterol was positive (Table 2), albeit after 14.8 months no more significant. Over the control period there were a negative development of serum triglyceride level and significant negative effects on the body weight. There were no complications among the 66 patients during the facultative ambulatory phase of the rehabilitation, in contrast to those 37 of 268 patients who had to be disclosed because of medical concerns (Table 1). During the rehabilitation the drug therapy was optimized for many participants. The control after 14.8 months revealed that these remedies were followed with great accuracy by the physicians at home (Figure 1). In conclusion, a cardiac rehabilitation program with an in-hospital start of 2 weeks, going over to ambulatory rehabilitation for 1 to 2 weeks, is as effective and safe as an in-hospital cardiac rehabilitation. As known, amelioration of the long-term results has yet to be achieved. PMID- 10372304 TI - [Ambulatory cardiac phase II rehabilitation--"the Cologne model"--including 3 year-outcome after termination of rehabilitation]. AB - From January 1992 until December 1994 the Cologne model of ambulant cardiac rehabilitation (ACR) in the greater area of Cologne, Germany, was performed and is still in progress. In Germany until 1992 the cardiac rehabilitation was exclusively performed stationary. The objective of the "Cologne model" was to evaluate, whether the transfer of the stationary cardiac rehabilitation programs into the ambulatory setting is achievable without deficits in efficiency, safety and overall quality. The results obtained are intended to serve for standardization and quality control of future ambulatory cardiac rehabilitation programs in Germany. From 1992 to 1994 108 patients (94 men, 14 women; 52.3 +/- 8.0 years old) with coronary artery disease (CAD) which were compatible with the criteria of the "Cologne model" (Table 1) participated in the 4-week ACR. The indications for inclusion into the ACR were in 74 cases a myocardial infarction (MI), in 34 cases CAD without MI, but with PTCA/stent-procedure (Table 3). Seven patients discontinued the ACR prematurely, 2 patients because of cardiovascular reasons. Reasons for the preference of the ambulatory over a stationary cardiac rehabilitation program were in 40.6% of the patients refusal of "hospital ambience", in 43.6% familiar or in 12.9% professional reasons. During the 4-week ACR patients participated in a mean of 72.9 +/- 6.7 hours of therapy (Table 4). As a result of the ACR exercise tolerance increased highly significantly (**) from 116.4 +/- 28.8 to 129.9 +/- 34.6 watt). This improvement was maintained at the 1- and 3-year control (128.7 +/- 35.8**) examinations (Tables 5 and 7). One year after ACR 77% of the patients stated to be physically active in ambulatory heart groups (AHG) (27.6%) or on their own (49.4%). Three years after ACR the rate of regularly physically active patients still was 59.2%. Furthermore, as a result of ACR the dietary behavior was changed significantly. There was a reduction in the consumption of lipids by 20.8%, saturated fatty acids by 30.7% and of cholesterol by 30.5%. The plasma concentrations of cholesterol decreased from 231 +/- 49.8 to 213.2 +/- 35.9 mg%**. Six (and 12) months after ACR they increased again to 225.6 +/- 39.4 mg%. Three years after ACR the mean cholesterol level was 219.1 +/- 39.3 mg%. In the high risk group (cholesterol at the initial visit > 220 mg%) cholesterol levels were reduced from 266 +/- 44 to 232 +/- 31.9 mg%**. Six and 12 months after ACR they were 239.7 +/- 35.8 mg% and 245.8 +/- 32.6 mg%, respectively, (Tables 6 and 7) and still significantly lower than before ACR, though only 19% of the patients were treated with lipid lowering agents. Three years after ACR cholesterol were 234.6 +/- 37.7 mg%** in the high risk group. 34.2% of the patients received lipid lowering agents. Mean body weight remained unaltered over the 3-year period. Smoking behavior was not altered significantly during the 4-week ACR. However, before the cardiovascular event 67.3% of the patients had smoked cigarettes. At the beginning and at the end of ACR 20.8% of the patients still smoked. During the ACR the number of smoked cigarettes was reduced significantly from 32.4 +/- 15.2 to 6.9 +/- 5.2 cigarettes per day. One year after ACR 23% of the patients were smokers, 3 years after ACR the percentage of smokers increased to 30.3%. Before ACR 73.3% of the patients were still working. During the first 6 months after ACR 68.2% returned to work and the percentage increased to 73% in the following 6 months. The results demonstrate that it is achievable to transfer the contents of the established stationary cardiac rehabilitation programs into the ambulatory setting without loss of efficiency, safety and overall quality. It is further confirmed, that it is necessary to continuously evaluate the results of the cardiac rehabilitation program on a long-term basis. (ABSTRACT TRUNCATED) PMID- 10372307 TI - [Experiences with ambulatory cardiologic rehabilitation]. AB - Since the founding of the Dusseldorf Outpatient Cardiologic Rehabilitation Center (Berliner Allee) in 1990 approximately 500 outpatient rehabilitation measures were performed, predominantly as follow-up treatment. The Center is currently participating in 2 pilot programs in outpatient cardiologic rehabilitation initiated by pension scheme providers and health insurance companies. A holistic approach to the concept of therapy is fundamental to an intensive rehabilitation of the heart patients. In conjunction with physiotherapeutic exercise, a health education program and extensive psychological care form the basis of the broader therapy program. An on-going implementation of measures designed to assure the maintenance of structural and procedural quality is fundamental to the creation of a positive subjective experience on the part of the person undergoing rehabilitation as well as the attainment of the individual's specified rehabilitation goals. PMID- 10372308 TI - [Outcome of a four-week ambulatory cardiac rehabilitation (phase II) on cardiovascular risk factors, physical fitness and occupational reintegration in patients after myocardial infarct, dilatation treatment and heart operation]. AB - From October 1994 to July 1996, 128 patients (30 women, 98 men) participated in an outpatient cardiac rehabilitation program (phase II). Our objectives were to demonstrate risk-factor modification and increased workload capacity resulting directly from the rehabilitation in terms of primary results and long-term effects 6 and 12 months (n = 118, Figure 1) respectively 1.5 and 2 years (n = 87) after termination of the program (Tables 9 to 12). We observed how many of the patients were able to be occupationally reintegrated after completion of phase-II rehabilitation. Workload capacity significantly increased from 1.2 W/kg upon entry to 1.5 W/kg (p < or = 0.05) upon completion of 4 weeks cardiac rehabilitation. Workload capacity remained consistently high at 6 months and 1 year (1.5 W/kg) and at 1.5 and 2 years (1.7 W/kg). Total cholesterol decreased significantly from 247 to 201 mg/dl (p < or = 0.05) during the 4-week program. Significant cholesterol (p < or = 0.01) reductions persisted at 6 months (216 mg/dl) and 1 year (215 mg/dl). After 1.5 and 2 years, the total cholesterol was less than 14% and 17% below the mean of cholesterol at the beginning of the program. Similarly, LDL cholesterol was 185 mg/dl before entering the program, 146 mg/dl after 4 weeks, 151 mg/dl after 6 months and 149 mg/dl after 1 year. Triglyceride levels showed a significant reduction (p < or = 0.01) with levels 189 mg/dl before entering the program, 148 mg/dl after 4 weeks, 151 mg/dl after 6 months and 154 mg/dl after 1 year. LDL cholesterol and triglyceride levels did not significantly increase after 1.5 and 2 years. The HDL cholesterol increased slightly as a long-term effect (from 51 mg/dl before entering the program to 55, 56 and 54 mg/dl after 1, 1.5 and 2 years, respectively). Seventy-three percent of the patients questioned (n = 73) found the program very good, 27% said it was good and no patient was dissatisfied. Fifty-one (81%) of the 63 patients who were actively employed before becoming ill and later entering our program were immediately able to be reintegrated into their previous occupation. In several cases reintegration took 7 weeks. Seven (11%) patients applied for pension, 5 (8%) patients remained unemployed on sick-leave. PMID- 10372309 TI - [Ambulatory/partial inpatient phase II rehabilitation of heart patients in a Rhine-Main district rehabilitation clinic]. AB - According to the guidelines and standards for a comprehensive phase II rehabilitation a new outpatient/part-time outpatient program was developed and started at an existing rehabilitation center in January 1997. All patients of the new program were included in a follow-up study and compared with patients of the same center, who met the inclusion and exclusion criteria for the outpatient mode, but wanted to perform their rehabilitation in the full-time residential mode. Both groups were examined before, at the end and 6 months after the program. Until December 1998 118 patients after an acute cardiac event such as myocardial infarction, PTCA or heart surgery were rehabilitated in the outpatient/part-time outpatient mode. The short- and medium-term results concerning somatic outcomes, e.g. the risk factor profile or the improvement of the maximum work capacity, were equal in both groups. Comparing the direct costs for a 4-week rehabilitation, the part-time outpatient program was 26%, the outpatient program even 52% cheaper than the standard full-time residential program. The new program is as effective as the residential rehabilitation, but it is cheaper. Because of the in- and exclusion criteria it is suitable for only a subgroup of cardiac patients, because of the demands and standards the new program it can be offered only in special rehabilitation centers. PMID- 10372310 TI - [Partial inpatient cardiologic rehabilitation in an urban satellite center of a rehabilitation clinic: the Munich model]. AB - In April 1996 the Munich Rehabilitation Center was founded by the Social Security Agency as a "satellite center" of the "Klinik Hohenried for Cardiovascular Diseases", which is located 50 km south of Munich near the lake Starnberg and performs in-patient rehabilitation since 1967. The Munich Rehabilitation Center is exclusively designated for outpatient rehabilitation for patients in the Munich area. Monday to Friday from 9 a.m. to 4 p.m. up to 50 patients are treated according to the same standards as in residential centers. About 40% of patients treated are in WHO phase II, e.g. after coronary artery bypass grafting, acute myocardial infarction or a percutaneous transluminal coronary angioplasty. 60% have a chronic stable cardiac disease or risk factors for arteriosclerosis. After almost 3 years of experience we see some specific advantages in outpatient compared to the in-patient setting. Ambulatory treatment seems to be more than the residential center "minus a bed". For example: a "holiday feeling" can be avoided by the location in a city area, so the patients are focusing on medical rehabilitation. Outpatient rehabilitation makes an easy transition from a cardiological center to every day life possible. Some patients would refuse any further treatment far away from home because of personal or occupational reasons. Anxiety can be reduced and self-confidence increased by the outpatient setting. There is a daily feedback about the ability to transfer therapeutic advices home. We learned to appreciate outpatient rehabilitation as a cost-effective supplement to the proven in-patient setting for patients in municipal areas. PMID- 10372311 TI - [Rehabilitation with the "Ennepetal model"]. AB - The rehabilitation model which is described in this paper shows that optimization of the goals of rehabilitation such as improvement of vocational reintegration is possible by means of a flexible rehabilitation process and that this can also be achieved using existing resources. The patients are cared for according to their disabilities and functional disorders along what can be described as a rehabilitation course, ranging from in-patient and outpatient/semi in-patient treatment to outpatient heart groups. It was possible to achieve marked improvements in vocational reintegration over a 2-year period by an intervention program (intensified aftercare treatment, INA). After completion of follow-up rehabilitation, the number of generally disabled pensioners was substantially lower in the intervention program than in the case of patients in the control group. It has also been shown clearly that flexible rehabilitation models require co-operation on the part of all those involved in the rehabilitation process. PMID- 10372312 TI - [Clinical follow-up 6 months after ambulatory/partial inpatient after-care rehabilitation. Further results of the Cologne model of ambulatory cardiac phase II rehabilitation]. AB - Three hundred and thirty patients with coronary artery disease (CAD) (288 men, 42 women, age of 55.5 +/- 10.0 years) participated in a 4-week ambulatory cardiac rehabilitation program (ACR) (Table 1). The cardiovascular indication for ACR was in 229 cases a myocardial infarction. In 101 patients a CAD with invasive revascularization but without a history of MI was present. In 92 patients with myocardial infarction additionally an invasive revascularization was performed. Eighty-three patients were included after a CABG-procedure (Tables 2 to 5). Six months after the ACR 290 (87.9%) patients presented for clinical reevaluation. In 235 (81.0%) of the 290 examined patients the cardiovascular diagnosis was unaltered. In the first 6 months after ACR in 76 (26.2%) patients a coronarography was performed, in 44 patients a restenosis was diagnosed. In 36 patients an additional invasive procedure (in 28 patients a PTCA, in 5 patients with additional stent-implantation, in 1 case with rotablation, in 8 patients CABG) was performed. In 1 patients a pace-maker was implanted. Since the ACR 1 patient experienced a myocardial infarction and 2 a recurrent myocardial infarction. In 1 patient myocardial fibrillation occurred. Totally, 70 patients (24.1%) required stationary-hospital treatment during the first 6 months after ACR (Table 6). In 11 cases an acute admission to hospital treatment because of cardiovascular reasons was documented. The majority of the hospital admission was elective, because of diagnostic or therapeutic procedures. In 6 patients a CABG surgery was performed. In approximately 80% of the patients the cardiovascular status was stable during the first 6 months after ACR. Though 24.1% of the patients required stationary hospital treatment, the majority of the admissions was elective of interest, there was a high rate of hospital admissions in the PTCA-group in combination with recoronarographies and revascularization because of early reocclusion. PMID- 10372313 TI - [Initial experiences with a comprehensive ambulatory rehabilitation program for heart patients]. AB - 213 patients (197 males and 16 females, mean age 58.5 years) have taken part during the first 5 years in our comprehensive outpatient cardiac rehabilitation program (101 patients after acute myocardial infarction, 73 patients after heart surgery and 39 patients with various cardiac problems). The program consists of three elements: (1.) regular physical training in an outpatient group three times a week for 6-12 weeks; (2.) information and education of the patient and if possible of the family about risk factors and life-style changes, and (3.) psychosocial support. During 10,838 hours of active rehabilitation, no life threatening complication occurred. After completion of the program, 73% of the patients were fully rehabilitated (return to work in the active working population, reintegration in the family and daily life in the retired and non working patients), 15% of the patients interrupted the program because of further invasive treatment, and in 11% the goal of the rehabilitation program was not achieved. Follow-up was carried out by a questionnaire 3-56 months (mean 25 months) later. The percentage of smokers decreased from 55% before the program to 18% (p < 0.05), regular physical exercise increased from 49% to 76% (p < 0.05), and the perceived stress level of high intensity decreased from 53 to 9% (p < 0.05). Our findings indicate that a comprehensive cardiac rehabilitation program in an outpatient setting in conjunction with a community hospital is feasible; it can be conducted with a high degree of safety and is followed by considerable lifestyle changes. PMID- 10372314 TI - [Comprehensive ambulatory rehabilitation for heart patients--1999 status]. PMID- 10372315 TI - [Ambulatory long-term rehabilitation of heart patients]. AB - In Germany cardiac rehabilitation contains a comprehensive 3 to 4 week inpatient program. The aim of our study was to perform an outpatient long-term rehabilitation including intense exercise and behavior therapy. In this setting the health benefits could be increased over the first 6 months. Hundred and twelve patients (94 men and 18 women, age 55 +/- 11 years) after myocardial infarction (52%), bypass-surgery (37%), PTCA (23%), and others (15%) were included in the ongoing study. Cholesterol and LDL-cholesterol diminished significantly. HDL-cholesterol was increased significantly after 6 months. The endurance exercise capacity per rate-pressure-product was increased by 46% during the 6 months period. The intake of cholesterol could significantly be diminished from 307 +/- 25 to 258 +/- 19 mg per day. Five of 16 patients became free from smoking. The first results from the long-term outpatient program show that the time of intervention and also the intensity of the medical, exercise and the behavior therapy are important factors for a successful rehabilitation. PMID- 10372316 TI - [Patient information and informed consent]. PMID- 10372317 TI - [Attitude: why this reticence with regard to patient information in medical imaging?]. PMID- 10372318 TI - [The treatment of osteoid osteoma: a multitude of choice: surgery, percutaneous resection, alcohol injection or thermocoagulation]. PMID- 10372319 TI - [Intravaginal ultrasonography of the uterine cervix: hope in the campaign against premature births]. AB - Transvaginal ultrasound of the cervix provides an objective and non-invasive method for assessing cervical status: cervical measurements, anatomy (wedge, "funneling" of the amniotic sac protruding into the internal os), functional examination (dynamic modifications of the internal orifice in response to uterine contractions or pressure on the fundus). In patients with symptoms (that is, presenting preterm labor), the positive predictive value of cervical ultrasound is far better than that of digital examination and allows better screening of high-risk patients; in addition, its negative predictive value is excellent, which should reduce unnecessary treatments related to prevention of preterm births (excessive tocolysis and unnecessary or prolonged hospitalization) without increasing the incidence of such births. In high-risk asymptomatic patients, ultrasound of the cervix has been proposed to reduce the number of cerclage in cases where the situation is unclear. Finally, in the general population, it could be added to the second trimester ultrasound, to improve screening of patients at risk of preterm delivery. PMID- 10372320 TI - [Apprenticeship in radiology at the University Hospital Center in Grenoble: research on the most effective methods]. AB - From analyses of strengths and weaknesses of the apprenticeship of radiology in France, of existing pedagogic methods, of time constraints within clinical settings and of scientific theories of teaching and learning, the authors define objectives that teaching should target and propose a new method of instruction problem and practise-based. The method appears applicable in French radiology departments. It should forster early acquisition by residents of professional attitudes and way of thinking, and make them active self-directed learners. PMID- 10372321 TI - [Patient information and iodized contrast media]. AB - Prior information to patients concerning the risks associated with intravenous injection of contrast media for diagnostic imaging is rarely performed in France. PURPOSE: Evaluate patients' desire for information about risks of intravenous injection of contrast material and its impact on their level of confidence. MATERIAL AND METHODS: Two hundred and twenty seven adult patients, while awaiting a CT scan with injection, read an information form reviewing the risks associated with intravenous injection of contrast material. They filled out an answer sheath concerning their desire to be informed and the impact of that information on their level of confidence. Two levels of risk were evaluated, one where the risk of death was included and one where the risk of death was not included. RESULTS: Eighty six percent of patients wished to be informed about the risks; eleven per cent felt they became more anxious after being informed, irrespective of the mention or not of the risk of death. CONCLUSION: Patients wish to be informed about the risks associated with the intravenous injection of contrast material. This information seems to create only mild anxiety. PMID- 10372322 TI - [Emergency computed tomography in a general hospital center]. AB - This retrospective study was performed to precise indications of emergency CT in a general hospital over a 30 months period. We tried to determine, with help of prior studies, the indications for CT of the brain in the management of acute meningitis, acute headache, and in the management of head injury. In acute meningitis, there is no evidence to recommend CT of the brain before lumbar puncture, except to identify patients at increased risk of cerebral herniation. The imaging study of choice in subarachnoid hemorrhage is non enhanced CT scan. This exam has to be performed in case of acute headache. The CT evaluation of patients with minor head injury remains controversial. PMID- 10372323 TI - [Computed tomography study of the sigmoid colon: discriminating diagnostic criteria and interobserver correlations]. AB - PURPOSE: To assess the value of pericolonic findings at CT in the evaluation of the sigmoid colon. MATERIALS AND METHODS: A total of 210 CT examinations were retrospectively reviewed by 3 blinded radiologists. Data was analyzed to determine the interobserver correlation and the value of pericolonic and colonic wall findings in diagnosis of sigmoid colon pathology. RESULTS: The interobserver correlation for pericolonic findings was equal to or superior to that for colonic wall findings. The presence of abnormal pericolonic fat was the most sensitive (88%) and specific (93%) sign to differentiate a diseased sigmoid colon from a normal one or from sigmoid diverticulosis. Wall-thickening was less sensitive (82%) and specific (76%). Findings suggesting malignancy over diverticulitis included acute zone of transition, focal fatty infiltration, and lymph nodes. Symmetrical and circumferential wall thickening, target-like enhancement, and local fatty proliferation were findings suggesting colitis over diverticulitis. Wall thickening more than 15 mm, involvement of 15 cm or less, asymmetrical involvement, acute zone of transition, and homogeneous or heterogeneous enhancement were findings suggesting malignancy over colitis. CONCLUSION: To render a diagnosis, the evaluation of the fat infiltration must prevail on the parietal thickening appreciation. PMID- 10372325 TI - [Ultrasonographic diagnosis of an epiphyseal detachment of the upper end of the humerus due to birth injury]. AB - Injuries to the proximal humerus in infants are often missed or misinterpreted because of the non ossification of the epiphysis. Ultrasound allows direct visualisation of the proximal humeral epiphysis, metaphysis, joint space, and relationship with the glenoid cavity. Ultrasound diagnosis of other epiphyses birth trauma has been reported. Ultrasound should be considered as the first imaging modality if traumatic epiphyseal dislocation is suspected in a newborn. PMID- 10372324 TI - [Percutaneous resection under computed tomography guidance of osteoid osteoma. Mid-term follow-up of 38 cases]. AB - PURPOSE: To evaluate the mid-term outcome following CT-guided percutaneous resection of osteoid osteoma. MATERIALS AND METHODS: 38 patients who had been treated by CT-guided percutaneous resection were included. The mean follow-up of 3.7 years. Early and mid-term outcome and histology were analyzed. RESULTS: Histological samples were adequate in 92% of cases and a diagnosis of osteoid osteoma was confirmed in 73.7% of cases. In 6 cases, the lesion was not an osteoid osteoma: 2 mucoid cysts, 1 benign fibrous dysplasia, 1 fibromucoid lesion, 1 focal osteochondritis, 1 osteomyelitis. Cure was achieved in 84.2% of patients. Minor transient complications occurred in 23.7% of cases. The most serious complications included: 1 intramuscular hematoma, 2 femoral fractures, and 1 case of S. aureus osteomyelitis. CONCLUSION: This study confirms that CT guided percutaneous resection of osteoid osteomas is effective and shows that other small lesions can also be treated using this technique. PMID- 10372326 TI - [Recurrent multifocal chronic osteomyelitis: scintigraphy or MRI. Apropos of 2 cases]. AB - We report two cases of chronic recurrent multifocal osteomyelitis. In both cases, MRI demonstrated the presence of asymptomatic lesions that the technetium bone scan failed to show. The asymptomatic lesions were located in the acetabelum and the distal femur. Despite the actual literature, MRI was more specific than bone scan in these two cases. PMID- 10372327 TI - [Percutaneous treatment of a superficial femoral artery aneurysm using an intravascular stent-prosthesis]. AB - One case of superficial femoral aneurysm treated percutaneously by endovascular stent graft (Passager Boston) is reported. The initial radiographic evaluation included arteriography and color doppler sonography which enable analysis of the flow path, the extent of the wall thrombus, the choice of stended graft size. The procedure of implantation was technically trouble free. The post-procedure 3D CT and arteriography demonstrated occlusion of the aneurysm and resaturation of normal flow path. The six and twelve month check confirmed the stability of the results locally and the integrity of run off vessels. In weakened and specially elderly patient percutaneous treatment of superficial femoral artery aneurysm can be carried out easily. The contribution of 3D CT is essential in follow up to ensure an optimal result and to detect any complication. PMID- 10372328 TI - [Ewing's sarcoma of the soft tissues: apropos of 3 cases and review of the literature]. AB - Three cases of extraosseous Ewing sarcoma are reported. This pathology of the young adult is very rare as shown by the review of the literature. Clinical or imaging (CT or MRI) findings are non-specific and diagnosis is based on histology. Nonetheless, this diagnosis should be considered in all patients with primary soft tissue tumors. PMID- 10372329 TI - [What is it? A narrow long-isthmus aortic coarctation with significant collateral circulation]. PMID- 10372330 TI - [Inforad 98]. PMID- 10372332 TI - [Pediatric radiology]. PMID- 10372331 TI - [Whole-body MRI]. PMID- 10372333 TI - [Neuroradiology]. PMID- 10372334 TI - [Thoracic imaging. "Not with grand innovation, but an equilibration between cross sectional imaging (chiefly computed tomography) and nuclear medicine"]. PMID- 10372335 TI - [Cardiovascular imaging. Diagnostic and interventional imaging]. PMID- 10372336 TI - [Breast imaging]. PMID- 10372337 TI - [Digestive imaging]. PMID- 10372338 TI - [Uroradiology]. PMID- 10372339 TI - [Osteo-articular imaging]. PMID- 10372340 TI - Relationship between youth and parent perceptions of family environment and social anxiety. AB - This study concurrently examined the relationship between adolescents' perceptions of their parents' child-rearing styles and family environment and their reports of social anxiety. Adolescents reporting higher levels of social anxiety perceived their parents as being more socially isolating, overly concerned about others' opinions, ashamed of their shyness and poor performance, and less socially active than did youth reporting lower levels of social anxiety. Parent perceptions of child-rearing styles and family environment, however, did not differ between parents of socially anxious and nonsocially anxious adolescents. Results are comparable to studies using adult retrospective reports and are discussed with regard to the role of the family environment in the development of social anxiety. PMID- 10372341 TI - Factor structure of social fears: The Liebowitz Social Anxiety Scale. AB - In the assessment of social anxiety, investigators often differentiate between social interactional anxiety and performance anxiety. The Liebowitz Social Anxiety Scale (LSAS), a clinician-administered measure of social anxiety and avoidance, was originally developed with separate subscales for the assessment of fear and avoidance of situations involving social interaction and performance/observation by others. Separate confirmatory factor analyses of the LSAS fear and avoidance ratings demonstrated that this two-factor model did not provide an adequate fit to the data, suggesting the need to further investigate the underlying structure of the LSAS. Separate exploratory common factor analyses of the fear and avoidance ratings yielded four similar factors for each: (1) social interaction, (2) public speaking, (3) observation by others, and (4) eating and drinking in public, which demonstrated convergent and discriminant validity with other measures of social anxiety. These findings suggest that there are four global categories of social fear assessed by the LSAS, and that while social interaction anxiety appears to be unifactorial, fear of performance/observation situations may be multifactorial. PMID- 10372342 TI - Multimodal comparisons of social phobia subtypes and avoidant personality disorder. AB - The purpose of the present study was to further clarify the behavioral, physiological, and verbal response of patients with circumscribed social (speech) phobia, generalized social phobia without avoidant personality disorder, and generalized social phobia with avoidant personality disorder. Patients completed a battery of verbal report instruments and participated in two behavioral assessment tests. Measures of avoidance/escape behavior, cardiac response, level of behavioral skill, state anxiety, and positive and negative self-statements during performance were collected. Significant differences across response domains were found between the circumscribed social phobia and the generalized groups. Most of the distinctions were between individuals with circumscribed social phobia and those with both generalized social phobia and avoidant personality disorder, with the former group having less overall psychopathology. In addition, there was substantial overlap of problems between generalized social phobia individuals with and without avoidant personality disorder. Implications for the conceptualization of social phobia are discussed in terms of the differences among social phobia subtypes. PMID- 10372343 TI - Temporal characteristics of evaluation anxiety. AB - The temporal characteristics of evaluation anxiety are not well-defined by previous research. We examined the effects of length of the pre-evaluation interval (3, 6, or 12 minutes) and stage of the pre-evaluation interval at which evaluation anxiety was measured (start, middle, or end) on evaluation anxiety while participants performed an activity that was the focus of the impending evaluation. Participants wrote their opinion on a controversial social issue while anticipating a subject matter expert's judgment of their social maturity, and evaluation anxiety was measured by a battery of state anxiety measures. Higher levels of evaluation anxiety were detected on the Worry-Emotionality Questionnaire (WEQ) Worry subscale at the end of the pre-evaluation interval than at earlier stages, regardless of interval length, although individual difference variables exerted an important influence. Individuals with high trait self presentation concerns experienced particularly high state anxiety (State-Trait Anxiety Inventory) at the start of the longest pre-evaluation interval. Low self efficacy individuals showed a U-shaped pattern across pre-evaluation stages on both the WEQ Worry and Emotionality subscales, while high self-efficacy participants showed either no change (worry) or an inverted-U pattern (emotionality). Implications for the experimental measurement of evaluation anxiety were discussed. PMID- 10372344 TI - Demographics, treatment seeking, and diagnoses of anxiety support group participants. AB - Two peer-led anxiety disorder support groups were surveyed to ascertain characteristics of individuals seeking the services of these groups. Both groups had received information and consultation from the Anxiety Disorders Association of America. One hundred and eighty-four individuals were interviewed for diagnosis by structured clinical interview; demographics and treatment-seeking behaviors were ascertained by self-report questionnaires. Both groups surveyed were composed of more females than males and were predominantly Caucasian. Treatment was most frequently sought from psychiatrists, psychologists, and family doctors. One fourth of the sample had sought help for anxiety in a hospital emergency room. Eighty-eight subjects (94%) at the Dallas site and 57 subjects (61%) at the Durham site met criteria for at least one current anxiety or affective disorder. More than half of those who met criteria for current panic disorder with agoraphobia also met criteria for at least one other anxiety disorder, or for major depression. Approximately one third of each support group met criteria for current social phobia. Severity of social phobia symptoms was assessed by four scales. An increased risk of substance abuse was noted for individuals with a diagnosis of social phobia, as compared with diagnoses of other anxiety disorders. PMID- 10372345 TI - Diagnostic viability of depressive personality disorder: theoretical and conceptual issues. AB - Depressive personality disorder (DPD) is being considered for inclusion in future editions of the Diagnostic and Statistical Manual of Mental Disorders (DSM). In this paper, we review the theoretical and empirical literature on DPD, and examine a number of issues concerning its viability as an Axis II disorder. Three criteria were used as necessary preconditions for the inclusion of DPD on Axis II: (a) substantial theoretical distinctiveness from other disorders, (b) established empirical distinctiveness from other disorders, and (c) sufficient deviation from a "normal temperament." We argue that it is improbable that this disorder could be diagnosed independently of dysthymia, and that there is also considerable symptom overlap with several Cluster C disorders. Although there appears to be sufficient deviation from normality among the entire group of individuals with DPD, those persons whose symptoms are not severe enough to cause overlap with dysthymia may not so deviate. We conclude that the current conceptualization of DPD does not have sufficient discriminative validity or clinical utility to warrant inclusion in future editions of the DSM. PMID- 10372346 TI - Depressive personality disorder: fact or fiction? PMID- 10372347 TI - Depressive personality traits and dysthymia: a commentary on Ryder and Bagby. PMID- 10372348 TI - Personality, disorder, and personality disorder: towards a more rational conceptualization. PMID- 10372349 TI - Selecting the most informative items in the IIP scales for personality disorders: an application of item response theory. AB - The first goal of the present analyses was to shorten the five scales (Pilkonis, P. A., Kim, Y., Proietti, J. M., & Barkham, M. [1996]. Journal of Personality Disorders, 10, 355-369) for personality disorders (PDs) developed from the Inventory of Interpersonal Problems (IIP), thereby increasing their attractiveness for screening purposes. The second goal was to illustrate, for more general purposes, the utility of item response theory (IRT) for such scale refinement. IRT analyses were performed using data collected from six different samples (N = 1149) at five sites and a two-parameter (2P) graded model designed for multiple response items like those on the IIP. The five most informative items from each scale were identified, based on the magnitude of item discrimination parameters and the range and elevation of individual item information functions. Preliminary analyses of the reliability and validity of the short forms of the scales (totaling 25 items) supported their value as alternatives to the longer forms (consisting of 47 items), although definitive tests of their psychometric properties await crossvalidation in independent samples. Analyses of the quality receiver operating characteristics (QROC) of the long and short forms showed that both versions can be useful in predicting the presence versus absence of any PD diagnosis arrived at by using either a "best estimate" clinical consensus method or a structured Axis II interview. PMID- 10372350 TI - Sadistic personality disorder in sex offenders: relationship to antisocial personality disorder and sexual sadism. AB - To investigate the relationship of sadistic personality disorder (SPD), as defined in the appendix of DSM-III-R, to other personality disorders and to sexual sadism, 70 sex offenders (27 child molesters, 33 rapists, and 10 murderers) were assessed by the International Personality Disorder Examination. In 19 subjects (27.2%) from the total sample, SPD was diagnosed. The highest overlap appeared with borderline personality disorder (31.6%) and antisocial personality disorder (42.1%). However, in four cases SPD was the only personality disorder diagnosed. Factor analysis of the antisocial and sadistic criteria resulted in four major factors--one factor with high loadings on the sadistic criteria and the violent criteria of antisocial personality disorder, two factors with different forms of adult and juvenile aggression, and a fourth factor with high loadings on the antisocial criteria covering exploitative behavior. The results do not support SPD as a discrete disorder. Nevertheless, SPD may be seen as an important subdimension of antisocial personality disorder, distinct from more exploitative forms of antisocial behavior with less violence. Of those patients with SPD, 42.1% also had a DSM-III-R diagnosis of sexual sadism, which may be the most dangerous configuration. PMID- 10372351 TI - Using the structural analysis of social behavior (SASB) to differentiate young adults with borderline personality disorder features. AB - The present experiment investigated the ability of the Structural Analysis of Social Behavior (SASB) Intrex questionnaires to differentiate nonclinical samples of young adults with features of either borderline personality disorder (BPD), schizotypal personality disorder (SPD), or no personality disorder (no PD). A number of significant differences were demonstrated on the SASB Intrex questionnaires when comparisons were made between the BPD and no PD samples. These results are consistent with those reported in an earlier investigation of BPD using SASB in a clinical sample. Few differences were found between the SPD features sample and the other two samples. These results are not consistent with previous findings that have differentiated BPD and SPD in clinical samples. The results are discussed in terms of the possible implications for SASB to differentiate BPD, the use of nonclinical samples in personality disorder research, and dimensional models for conceptualizing personality disorders. PMID- 10372352 TI - Preparation of nucleosome core particle from recombinant histones. PMID- 10372353 TI - Assembly of defined nucleosomal and chromatin arrays from pure components. PMID- 10372354 TI - Isolation of minichromosomes from yeast cells. PMID- 10372355 TI - Chromatin assembly in Xenopus extracts. PMID- 10372356 TI - In vitro reconstitution of nuclei for replication and transcription. PMID- 10372357 TI - Preparation/analysis of chromatin replicated in vivo and in isolated nuclei. AB - This article outlined biochemical methodologies for the labeling, detection, and analysis of newly replicated and newly assembled nucleosomes. The isolation of specific vertebrate factors that may be involved in chromatin assembly in vivo, such as nucleoplasmin, CAF-1, and NAP-1 and their counterparts in Drosophila and yeast add a further dimension to the study of nucleosome assembly in living cells. In particular, the ability to genetically manipulate the yeast system, together with the identification of yeast enzymes that acetylate newly synthesized H4, will certainly provide exciting new avenues for the investigation of chromatin assembly in vivo. PMID- 10372359 TI - Reconstitution of high mobility group 14/17 proteins into nucleosomes and chromatin. PMID- 10372358 TI - Interactions of high mobility group box proteins with DNA and chromatin. PMID- 10372360 TI - Purification and assays for high mobility group HMG-I(Y) protein function. PMID- 10372361 TI - Cryoelectron microscopic analysis of nucleosomes and chromatin. PMID- 10372362 TI - Electron microscopy of DNA-protein complexes and chromatin. AB - This article focused on a number of aspects of the preparation of chromatin and other DNA-protein complexes for conventional transmission EM that are critical for success but may not have been addressed in a single chapter before. These include the importance of optimizing fixation, the generation of active supporting supports, and the use of negative staining as a means of obtaining higher resolution detail than can be garnered from shadow casting methods. PMID- 10372363 TI - Targeted cross-linking and DNA cleavage within model chromatin complexes. PMID- 10372364 TI - Base-pair resolution mapping of nucleosome positions using site-directed hydroxy radicals. PMID- 10372365 TI - Localization of specific histone regions on nucleosomal DNA by cross-linking. PMID- 10372366 TI - Restriction enzymes as probes of nucleosome stability and dynamics. PMID- 10372367 TI - Nucleoprotein gel assays for nucleosome positioning and mobility. AB - Recent recognition of the sensitivity of polyacrylamide gel electrophoresis to macromolecular conformation has provided a source of new applications. In chromatin research, nucleoprotein gel electrophoresis can yield a direct and visual estimate of the number and relative abundance of different positions adopted by the core histone octamer on DNA, as well as their locations relative to the middle of the DNA fragment. It is the only technique available for the fractionation of such nucleosome positioning isomers and leaves them intact. Thus this simple method constitutes a powerful tool to analyze and manipulate populations of variously positioned nucleosomes in their native state. Complementing conventional invasive enzymatic procedures that rely on the analysis of cutting patterns on nucleosomal DNA, these procedures are now revealing that histone octamers can reconstitute to a number of discrete, often overlapping, locations on most DNA sequences. Further capitalizing on these advantages of nucleoprotein gel analysis, the development of the technique into a two-dimensional assay has permitted a rare view at the dynamics of nucleosome positioning. Nucleosomes can redistribute between possible positions on DNA, with the distribution patterns of nucleosomes along the DNA being in dynamic equilibrium at 37 degrees in relatively low ionic strength conditions. This mobility of nucleosomes on DNA means that possible positions of nucleosomes can be defined precisely but that the actual locations of the nucleosomes are dynamic. It provides a compelling argument that a nucleosome position should be regarded as a probability rather than a static factor type of binding. This supports a more dynamic view of the nucleosomal organization, which seems more in accordance with the dynamic nature of gene expression. In providing the flexibility for adaptation, multiple positioning and nucleosome mobility could constitute essential ingredients of the mechanisms by which chromatin participates in gene regulation. PMID- 10372369 TI - Assays for chromatin remodeling during DNA repair. PMID- 10372368 TI - Assays for interaction of transcription factor with nucleosome. PMID- 10372370 TI - Nuclear run-on assays: assessing transcription by measuring density of engaged RNA polymerases. PMID- 10372371 TI - Mapping chromatin structure in yeast. PMID- 10372372 TI - Assays for nucleosome positioning in yeast. PMID- 10372373 TI - Mapping DNA interaction sites of chromosomal proteins using immunoprecipitation and polymerase chain reaction. PMID- 10372374 TI - Immunological analysis of yeast chromatin. PMID- 10372375 TI - DNA methyltransferases as probes of chromatin structure in vivo. PMID- 10372376 TI - Mapping cyclobutane-pyrimidine dimers in DNA and using DNA-repair by photolyase for chromatin analysis in yeast. PMID- 10372377 TI - Analysis of Drosophila chromatin structure in vivo. PMID- 10372378 TI - Ultraviolet cross-linking assay to measure sequence-specific DNA binding in vivo. PMID- 10372379 TI - DNA-protein cross-linking applications for chromatin studies in vitro and in vivo. PMID- 10372380 TI - Chromatin immunoprecipitation assays in acetylation mapping of higher eukaryotes. PMID- 10372382 TI - Guanine-adenine ligation-mediated polymerase chain reaction in vivo footprinting. AB - The analysis of functional DNA regulatory sequences involved in transcriptional control is critical to establishing which proteins mediate cell-specific gene expression. The organization of erythroid LCRs is complex, consisting of multiple, interdigested cis elements. As in situ binding to these sites is determined by the accessibility of these regulatory regions in native chromatin and the availability of relevent cell-specific and ubiquitous factors, in vivo footprinting was used to define protein DNA interactions in human globin LCRs. To further enhance the detection of protein contacts with this technique, we have modified the dimethyl sulfate-based ligation-mediated PCR in vivo footprinting procedure to permit the assessment of protein binding at guanine and adenine resides, rather than exclusively at guanines. This modification, termed GA-LMPCR in vivo footprinting, was essential for the analysis of GATA-1 motifs in the alpha-LCR and HS-3 of the beta-LCR. Moreover, GA-LMPCR in vivo footprinting provided high-resolution analysis of AP-1/NF-E2 elements and revealed protein contacts at sequences that are not coincident with previously described regulatory motifs. A comprehensive discussion of this modification and sample illustrations from our studies have been presented to demonstrate the enhanced detection and resolution obtained with this procedure. PMID- 10372381 TI - Chromatin structure analysis by ligation-mediated and terminal transferase mediated polymerase chain reaction. PMID- 10372383 TI - Exonuclease III as a probe of chromatin structure in vivo. PMID- 10372384 TI - Protein image hybridization: mapping and positioning DNA-protein contacts along DNA. PMID- 10372385 TI - In vivo analysis of chromatin structure. PMID- 10372386 TI - Analysis of nucleosome positioning in mammalian cells. PMID- 10372387 TI - Measurement of localized DNA supercoiling and topological domain size in eukaryotic cells. PMID- 10372388 TI - Fluorescence in situ hybridization analysis of chromosome and chromatin structure. PMID- 10372389 TI - Analysis of nuclear organization in Saccharomyces cerevisiae. PMID- 10372390 TI - Histone acetyltransferases: preparation of substrates and assay procedures. PMID- 10372391 TI - Analysis of activity and regulation of hGcn5, a human histone acetyltransferase. PMID- 10372392 TI - Purification of a histone deacetylase complex from Xenopus laevis: preparation of substrates and assay procedures. PMID- 10372393 TI - Purification and biochemical properties of yeast SWI/SNF complex. PMID- 10372394 TI - Analysis of modulators of chromatin structure in Drosophila. PMID- 10372396 TI - [Angiogenesis]. PMID- 10372395 TI - Purification of Drosophila nucleosome remodeling factor. PMID- 10372397 TI - [Emergence of the endothelial network during embryonic development]. AB - The avian model provides an experimental approach for dissecting the origin, migrations, and differentiation of cell lineages in early embryos. In this model, the endothelial network was shown to stem from both the somites and the splanchnopleural mesoderm. The somite line age produces only endothelial cells, whereas the splanchnopleural line age also produces hematopoietic stem cells. Potentialities of the mesoderm are determined by a positive influence from the endoderm and a negative influence from the ectoderm. A novel mode of blood-borne angiogenesis is also described. PMID- 10372398 TI - [Endothelial cell precursors in the avian embryo]. AB - Whereas the origin and migration of endothelial cells (ECs) have been studied primarily in the avian embryo, the molecular mechanisms governing these events in birds and mammals were unraveled following the identification of specific growth factors and of their receptors. In particular, analytic and experimental studies of vascular endothelial growth factor (VEGF) and its receptors have provided significant insights into the developmental biology of the vascular system. VEGFR2 is the earliest marker expressed by EC precursors in chickens and mice. Based on the localization of VEGFR2-positive cells in the avian embryo and on findings from clonal culture experiments, two types of EC precursors can be differentiated as early as the gastrulation stage, namely posterior mesoderm hemangioblasts capable of differentiating into both ECs and hematopoietic cells and anterior angioblasts capable of yielding only ECs. PMID- 10372399 TI - [Interactions between steroidogenic cells and capillary endothelial cells in the adrenal gland]. AB - Adrenocortical capillary endothelial (ACE) cells showed a two- to three-fold increase in number when they were cocultured with bovine adrenocortical (BAC) cells from the zona fasciculata-reticularis. This effect was detectable within 48 h, persisted throughout the seven-day coculture period, and occurred in the absence of addition of exogenous growth factors. It was similar to that obtained by addition of 1 ng/ml of FGF-2. Adrenal cortex tumor cells from humans (NCI) and mice (Y1) also stimulated ACE cell growth, whereas NIH 3T3 cells did not, suggesting an effect specific of steroid-producing cells. Addition of an FGF-2 neutralizing antibody failed to inhibit the BAC cell-induced ACE cell growth stimulation, indicating that FGF-2 was not involved. BAC cell-conditioned medium stimulated ACE cell growth, indicating a role for a diffusible factor. When BAC cells were cocultured with ACE cells in a 3D collagen gel, capillary-like tubes developed, consistent with secretion by BAC cells of angiogenic factors. Studies are under way to identify these factors. PMID- 10372400 TI - [Activation of latent TGF-beta. A required mechanism for vascular integrity]. AB - Recent molecular genetics studies in humans and mice showed that transforming growth factor-beta (TGF-beta) is involved in vasculogenesis and maintenance of blood vessel integrity. These results confirm earlier in vitro studies demonstrating generation of active TGF-beta when endothelial cells are cocultured with smooth muscle cells or pericytes. TGF-beta is secreted as a latent, inactive complex and becomes active only when released. Latent TGF-beta binds covalently to proteins (LTBP) that target it to the extracellular matrix. Thus, the latency of TGF-beta is essential to the regulation of the bioavailability and activity of this cytokine. The development of methods for measuring activation of latent TGF beta in cell cultures and identification of the proteins contained in the latent TGF-beta complex have shed new light on the mechanism of activation of latent TGF beta possibly involved in vasculogenesis, angiogenesis, and other processes. PMID- 10372401 TI - [Expression of a new family of receptors similar to CXC chemokine receptors in endothelial cell precursors]. AB - Characterization of a new family of G protein-coupled receptors is reported. Expression of these receptors is associated with endothelial lineage. Cloning of the Xenope X-msr receptor allowed to show that embryonic expression of this receptor occurred in the heart and developing primary blood vessels. Furthermore, within these cardiovascular structures, expression was restricted to the endothelial layer. Because structural similarities with the human orphan receptor h-APJ were found, the msr/apj receptor was cloned in mice. This showed that embryonic expression of this receptor was also confirmed to endothelial precursors. Thus, this receptor is the orthological equivalent in mice to the amphibian receptor X-msr. Molecular phylogenesis studies showed that the X-msr, msr/apj, and h-APJ receptors shared considerable homology with two CXC chemokine receptors, namely LCR1, whose name was recently changed to CXCR4, and RDC1, which is structurally similar to the CXCR2 receptor. The human h-APJ receptor is a co receptor for entry of the HIV into T cells, a property associated only with CXC chemokine receptors in the lymphocyte population. These data suggest that this new signaling system may participate in endothelial precursor migration during developmental angiogenesis and in endothelial cell migration and proliferation during neoangiogenesis in adults. PMID- 10372402 TI - [Thrombospondins: multimodular proteins with angiostatic function]. AB - The thrombospondins (TSPs) are a family of multimodular proteins that bind to the extracellular matrix with strong affinity. Of the five members of the TSP family, TSP1 and TSP2 are the only ones that inhibit endothelial cell migration in vitro and neoangiogenesis in vivo. This angiogenesis-inhibiting effect is mediated by interaction of a short sequence in type I modules with the membrane receptor CD36. TSP1 and TSP2 gene knockout experiments in mice showed increased blood vessel density in TSP2 -/- but no such alteration in TSP1 -/- animals. Loss of TSP1 gene expression was correlated with acquisition of an angiogenic phenotype in several models of human malignant tumors. Taken in concert, these findings suggest that TSP1 and, to a lesser extent, TSP2 may have therapeutic potential as angiogenesis-inhibiting factors. PMID- 10372403 TI - Angiogenin, a potent mediator of angiogenesis. Biological, biochemical and structural properties. AB - Angiogenin is a heparin-binding 14 kDa plasma protein with angiogenic and ribonucleolytic activity. Although its structural features have been extensively studied, an understanding of its physiological role and of how its properties are expressed continues to elude researchers. This article discussed some of the biological, biochemical, and structural properties of angiogenin. PMID- 10372404 TI - [Implication of HARP in angiogenesis and possible therapeutic role]. AB - HARP (heparin affin regulatory peptide), also called pleiotrophin (PTN), belongs to the heparin binding growth factors (HBGFs) family. Several new data suggest a role for HARP during the various stages of angiogenesis. In vivo, HARP is localised in endothelial cells of blood capillaries. In vitro, HARP displays mitogenic activity on endothelial cells, induces the formation of capillary-like structures in collagen gel, and degrades extracellular matrix via stimulation of plasminogen activator activity. HARP is also involved in neoangiogenesis during tumor progression. This review discusses the possible role of HARP in tumor angiogenesis and its therapeutic implications. PMID- 10372405 TI - [Transcription factors and angiogenesis]. AB - Various strategies led to the identification of transcription factors that take part to the control of different steps during the formation of new blood vessels: the description of the expression pattern of genes encoding these factors during embryonic development for ETS-1, ERG and FLI, SCL/TAL, GATA1 and 2, the description of the phenotype of embryos obtained after gene inactivation by homologous recombination for ARNT or LKLF, and the study of transcriptional regulation in cultured endothelial cells for EGR1 or HOX-D3. Altogether, these results showed that there is no transcription factor specific for endothelial cells or for one step in the formation of blood vessels. Rather, factors controlling gene expression induced by hypoxia, shear-stress or growth factors take part in the morphogenesis of the vascular tree. The study of these factors may allow to identify potential therapeutic targets for treatments aimed at inhibiting or stimulating the development of new blood vessels. PMID- 10372406 TI - [Role of fibroblast growth factor-2 in tumor angiogenesis]. AB - Angiogenesis reflects a balance between stimulating and inhibitory factors, among which fibroblast growth factor-2 (FGF-2) and vascular endothelial growth factor (VEGF) play a central role. FGF-2 stimulates endothelial cell growth and migration and enhances angiogenesis, both in vitro and in vivo. FGF-2 binds to a specific receptor (FGF R1) at the surface of endothelial cells. In addition to its extracellular effects, FGF-2 exerts intracellular effects that seem to be independent from tyrosine kinase-type receptors. FGF-2 is produced by several tumor types, including brain tumors, skin tumors, and fibrosarcomas. FGF-2 may play a pivotal role in angiogenesis in these tumors. PMID- 10372407 TI - [Thrombospondins, tumor angiogenesis and breast cancer]. AB - Thrombospondin-1 and -2 are extracellular matrix proteins that are overexpressed in breast cancer tissue. Their role in breast cancer remains unknown. This article reviews the potential effects of thrombospondin-1 and -2 in breast cancer tumori genesis and metastatic dissemination. PMID- 10372408 TI - [Impact on tumor angiogenesis and tumor progression of expression of the 18 kd and 24 kd isoforms of FGF-2]. AB - The role of FGF-2 in tumor progression and tumor cell invasiveness was investigated using the rat bladder carcinoma cells NBT-II, which do not constitutively express FGF-2 or its membrane-spanning receptor. The NBT-II cells were transfected using expression vectors encoding either the 18 kD or the 24 kD isoform of FGF-2. The 24 kD isoform contains a nuclear localization signal. The transfected NBT-II cells that expressed 18 kD FGF-2 produced and secreted this factor as the biologically active form and retained an epithelial morphology. When injected to nude mice, the tumorigenic potential of these cells was not increased over that of non-transfected NBT-II cells; however, although the time to tumor development was long, the tumors were highly vascularized, indicating secretion of the angiogenic factor FGF-2. The transfected NBT-II cells that expressed 24 kD FGF-2 varied in their morphological appearance and did not secrete FGF-2; immunofluorescence and Western-blot studies showed that the FGF-2 was mainly intranuclear. When injected to nude mice, these cells produced tumors and migrated not only to the lymph nodes but also to the lungs where they produced metastases. In aggregate, these data indicate that stimulation of angiogenesis is not sufficient to increase tumor growth and that nuclear FGF-2 acts as a tumorigenic and metastasis-promoting factor in the NBT-II carcinoma model. PMID- 10372409 TI - [VEGF in therapeutic coronary and peripheral artery angiogenesis]. AB - An overview is presented of the various steps of angiogenesis, to which an important contributor is Vascular Endothelial Growth Factor (VEGF), a substance capable of increasing both the permeability and the mitosis rate of endothelial cells. VEGF is also a potent vasodilating agent. The beneficial effects of these properties on the ischemic vasculature have been documented in numerous animal models, leading to the concept of angiogenesis-enhancement as a therapeutic tool. The first clinical trials are under way. This area is discussed based on a literature review. PMID- 10372410 TI - [Angiogenesis and neuropathology]. AB - Angiogenesis is crucial for both tissue development and tissue function and is an integral part of central nervous system embryogenesis and tumor growth. Angiogenesis is extremely active during development, then remains stable during adulthood and decreases gradually during aging. Physiological processes and inflammation can transiently stimulate angiogenesis in adults. Angiogenesis is modulated by a host signaling pathways, growth factors, growth factor receptors, and membrane proteins associated with these receptors or transcription factors. In disorders affecting the central nervous systems, as in those arising elsewhere in the body, angiogenesis can become inadequate, excessive, or qualitatively abnormal (dysplasia). These abnormalities can result in cerebral trophic disorders and in secondary remodeling. A few examples are given to illustrate various types of primary angiogenesis disorders responsible for cerebral lesions and of secondary angiogenesis disorders caused by an underlying cerebral disorder. PMID- 10372412 TI - A molecular to molar analysis of communicative and problem behaviors. AB - Few studies have examined the relationship between communicative and problem behaviors that are already present in a behavioral repertoire. In this study, a detailed microanalysis of the antecedents and consequences of aggressive and communicative behavior of a 7-year-old boy was conducted. By using both descriptive and experimental methodologies, the data suggested that problem and communicative behavior were maintained on thin concurrent schedules of social negative reinforcement. A molar analysis of the descriptive data showed that the relative amount of time allocated to each behavior was a function of the relative amount of reinforcement that each behavior accrued. The implications of these findings are discussed in terms of conducting descriptive analyses and for enhancing the efficacy of interventions for problem behavior. PMID- 10372411 TI - A preliminary comparison of guided compliance and high-probability instructional sequences as treatment for noncompliance in children with developmental disabilities. AB - The efficacy of guided compliance and high-probability instructional sequences was compared with two children referred to an outpatient clinic for treatment of noncompliance. Parents were taught to implement the procedures in their homes, and parent-training outcomes for the two interventions were compared in terms of treatment effectiveness, procedural integrity, and parent satisfaction. Levels of compliance were higher under guided compliance than under high-probability instructional sequences. Nevertheless, parents rapidly learned to implement both treatments with a high degree of accuracy and reported equal satisfaction with the procedures. PMID- 10372413 TI - Problem social behavior in the workplace: an analysis of social behavior problems in a supported employment setting. AB - The social skills problems that may influence the work-related success of supported employees has been only infrequently documented in the research literature. Though a multitude of research describes the performance-related challenges faced by supported employees, few papers address the interpersonal difficulties encountered by supported employees in the workplace. The present paper uses job trainer or "coaches" logs and two promising rating scales (the Psychopathology Instrument for Mentally Retarded Adults [PIMRA and PIMRA-S]) to describe the social problems encountered by some supported employees. Job coach's logs indicated that approximately 58% of supported employees had experienced one or more incidents of interpersonal difficulty during their employment tenure and that 40% of the problems experienced by these individuals could be described as sexuality-related. Overall, about 25% of all supported employees had reported incidents of conflict with employees or customers that seemed sexuality-related. In addition, social and developmental factors that might contribute to the interpersonal problems found in the present research are discussed. PMID- 10372414 TI - An evaluation of functional variables affecting severe problem behaviors in adults with mental retardation by using the Questions about Behavioral Function Scale (QABF). AB - We examined the functions of five severe problem behaviors in a sample of 417 institutionalized persons with mental retardation by using the Questions About Behavior Function Scale. The behaviors we examined included self-injurious behavior, aggression, stereotypies, pica, and rumination. The most common function for all behaviors except aggression was nonsocial. Aggression, however, was maintained by external environmental contingencies. Particular items of the Questions About Behavior Function Scale were identified as more frequently occurring and critical in ascertaining behavioral function. Implications of these results for developing more effective treatment plans are discussed. PMID- 10372415 TI - Urban health: an ecological perspective. AB - At the Second European Conference on Environment and Health held in Helsinki in June 1994, urban health was attributed a high priority. This decision by the Ministers of Environment and Health from European countries reflects and reinforces a growing worldwide concern in the 1990s about the health status of residents of urban areas in all continents. The reasons for this concern include the rapid rate of urbanization and the increasing number of environmental, economic, and social problems, which have a negative impact on health and well being in cities. This review presents a theoretical and methodological framework for the study of this vast and complex subject area. The paper proposes and illustrates an ecological perspective by discussing housing conditions and homelessness, as well as the concentration of poverty and deprivation in precise neighborhoods. To promote health and well-being more effectively, the ecological perspective presented in this paper can be applied by public health officers, by urban planners, and by policy decision-makers at national and local levels to promote our understanding of the multi-dimensional nature of health disorders of citizens. This approach should be a high priority for the beginning of the 21st century. PMID- 10372416 TI - Asbestos toxicity: an immunologic perspective. AB - Asbestos has long been associated with a number of life threatening pulmonary diseases, including asbestosis and mesothelioma. While the lung is the primary target organ for asbestos toxicity, a number of clinical and experimental studies over the past 30 years have shown that the immune system may also be altered by exposure to asbestos at occupationally relevant concentrations. Whereas early clinical studies generally focused on systemic observations of immune alteration, more recent studies have assessed the immunological changes occurring in the lung, the primary target organ of asbestos. This review will focus on the investigations that examine the influence of asbestos on systemic and local immunity, as well as the role that the immune system may play in asbestos-related disease. PMID- 10372418 TI - The use of epidemiological data in the control of foodborne viruses. AB - The Codex Committee on Food Hygiene has recommended the adoption of Hazard Analysis and Critical Control Point (HACCP) as the basis for food safety control. To provide an objective basis for the construction of HACCP systems, epidemiological data are required. The data should be accurate, up-to-date, and identify emerging pathogens, such as viruses. The number of laboratory reports of small, round-structured viruses in England and Wales has increased from 400 cases in 1990 to 2387 in 1996. Although a food vehicle is not essential for the spread of viral particles, food my be the primary unidentified vehicle. The Advisory Committee on the Microbiological Safety of Foods recommends the use of the Kaplan Criteria, which can give strong circumstantial evidence that an outbreak is attributable to small, round-structured viruses. The application of these criteria would give a more accurate reflection of the involvement of viruses in the incidence of foodborne disease. This review considers the use of epidemiological data to support HACCP and risk-assessment systems. It discusses the implications of focusing on traditional pathogens, for example Salmonella spp., as opposed to emerging pathogens, for the design of control systems. Recommendations are made for improving the system of data collection. PMID- 10372417 TI - Nutrition, nutritional habits, obesity, and prevalence of chronic diseases in workers. AB - The purpose of the present investigation is to reveal the specifics of the nutrition, nutritional behavior (habits), the prevalence of obesity and of certain chronic diseases among workers. The subjects were 264 workers (203 males and 61 females) from the ammonium production department of a fertilizer plant, divided into two age groups: under and over 30 years of age. The data were collected by means of a food-frequency questionnaire about daily nutrition and the average quantity of food. The nutritional status was assessed on the basis of BMI. All workers underwent clinical examinations conducted by a range of different experts, including an internal diseases specialist, a neurologist, a cardiologist, an opthalmologist, an otorhino-laryngologist, and a dermatologist. Twenty hematological and biochemical indicators in blood and serum were measured. Assessment of the individual energy intake showed that hyperenergetic nutrition was typical of 67% of workers because of extra intake of fat, which was seen in 87.9% of all individuals examined. The daily fat intake of over 40E% was typical for almost half the females (45.9%). All age and gender groups displayed hyperprotein nutrition with pronounced cellulose (fiber) deficit and a high daily sodium intake. The frequency of overweight individuals (BMI = 25, 1-30 kg/m2) was 43.9%, whereas that of obese individuals (BMI = > 30 kg/m2) was 23.1%. A total of 67% of workers had excessive body mass. The hypertension prevalence rose significantly from 6.9% in Group I to 34.5% in Group II, and to 57.4% in Group III. Coronary heart disease was rare, but the seven cases registered were among the overweight workers. The radiculitis prevalence among workers with normal body mass was two-fold lower in comparison with both groups (overweight and obesity). We conclude that hyperenergetic and unbalanced nutrition is a factor that determines the prevalence of overweight and obesity. A significantly higher percent of overweight and obese workers suffered from hypertension, liver disease, diabetes, coronary heart disease, and eye-vessel diseases. A tendency toward rising radiculitis and musculoskeleton system disease prevalence was seen that parallels the increase in BMI. PMID- 10372419 TI - Pesticide exposure: human cancers on the horizon. AB - Dichlorodiphenyltrichlorethane, a halogenated hydrocarbon, was introduced as an insecticide in the 1940s. In her book "Silent Spring", Rachel Carson expressed her concern for the environment, plants, animals, and human health about the potential harmful effects of such chemicals. In 1972, the Environmental Protection Agency banned the chemical in the USA. DDT and its metabolite DDE are lipid soluble compounds that persist in the environment and bioaccumulate in the body in adipose tissue at levels far higher than those in blood and breast milk. This paper evaluates the possibility of cancer occurring in humans from DDT exposure. Some risk of lymphoma, leukemia, pancreatic cancer, and breast cancer was found in humans exposed to DDT. Animal studies showed a significant association between DDT administration and lymphoma, respiratory cancer, liver cancer, and estrogenic effects on mammary tissue. On the basis of on epidemiological principles, human studies were deficient in adequate sample sizes and were not exempt from such confounding factors as multiple chemical exposure, lifestyle factors, genetic, and other environmental influences. Extrapolation of data on DDT toxicity from animals to humans has limitations. With the persistence of DDT and DDE in the environment, the potential risk to the health of man, animals, and the environment remains. PMID- 10372420 TI - [Udder health on dairy farms. 1. Results of a longitudinal study on 300 Dutch farms]. AB - Udder health was studied in 300 dairy herds grouped in three categories according to the bulk milk somatic cell count. In all herds, lactating cows were housed in a free-stall barn during the winter. All herds participated in a three or four weekly milk recording system, had annual production quota between 300,000 and 900,000 kg, and were stocked with cows of the Holstein-Friesian or Dutch Friesian breeds. The incidence of clinical mastitis was not different among herds with a low (< or = 150,000), middle (151,000 to 250,000), or high (251,000 to 400,000 cells/ml) bulk milk somatic cell count. Clinical mastitis caused by Gram-negative pathogens occurred more often in herds with a low bulk milk somatic cell count. Clinical mastitis caused by Staphylococcus aureus, Streptococcus dysgalactiae, or Streptococcus agalactiae occurred more often in herds with a high bulk milk somatic cell count; however, the incidence of clinical Streptococcus uberis mastitis was not different among the three herd categories. The differences in bulk milk somatic cell count between the categories could be explained by the management practices studied. The incidence of clinical E. coli mastitis was mostly related to housing, hygiene, and milking machine. The incidence of clinical Staphylococcus aureus mastitis was associated with factors that were related to the bulk milk somatic cell count and to factors of which it was not clear whether they were a cause or effect of the high incidence of Staphylococcus aureus mastitis. The incidence of clinical Streptococcus dysgalactiae mastitis was related to nutrition, milking technique, and milking machine. The incidence of clinical Streptococcus uberis mastitis was associated with housing, nutrition, and milking machine. Two groups of farmers and herds could be differentiated. The first group was identified as 'Clean and Accurate', and the second group as 'Quick and Dirty'. The relationship between these two groups and bulk milk somatic cell count category was high. However, the relationship between the two groups and the incidence of clinical mastitis was weak. PMID- 10372421 TI - [Udder health on dairy farms. 2. Mastitis prevention programs]. AB - Herd-specific mastitis control programmes are based on the continuous monitoring of primary parameters, of which the bulk milk somatic cell count and the distribution of pathogens responsible for clinical and subclinical mastitis are the most important ones. When a mastitis control programme is started, the herd situation should be analysed. Sampling milk from cows with clinical and subclinical mastitis is essential. During the last 5 years, a number of programmes related to udder health and milk quality have been started in the Netherlands. These programmes are: 1) 'Chain Quality Milk', 2) a herd-specific Mastitis Planner, 3) a sampling programme for subclinical mastitis, and 4) a combined analysis of milk recording data, and subclinical and clinical mastitis data. Using these four programmes, farmers, veterinary practitioners, and other advisors can design a herd-specific programme to improve udder health. Although there have been many studies on mastitis and important progress has been made, some important aspects have often not been studied. Suggestions for further research are made. PMID- 10372422 TI - [Urolithiasis in dogs and cats. Meeting of the Panel for Nutrition and Dietetics of the Veterinary Society]. PMID- 10372423 TI - Pregnancy and hypoxia. PMID- 10372424 TI - Electroencephalography and magnetic resonance imaging after diving and decompression incidents: a controlled study. AB - Electroencephalography and magnetic resonance imaging after diving and decompression incidents: a controlled study. Undersea Hyper Med 1999.; 26(2):61 65.--Diving incidents with symptoms of decompression sickness (DCS) and/or arterial gas emboli (AGE) might increase the degree of pathologic change in the electroencephalogram (EEG) or magnetic resonance imaging (MRI) of the supraspinal central nervous system (CNS). Diving itself, even without known symptoms of DCS and/or AGE, has been proposed to increase the number of CNS lesions using either EEG or MRI. In the first part of a two-part study we examined the effects of recompression treatment on EEG in decompression incidents in a group of sport and professional divers compared with a control group of healthy naval divers. In the second part we recorded brain MRI from three groups of volunteers: 1) divers who were treated for DCS in pressure chamber, 2) divers who had never had symptoms of DCS (and/or AGE), and 3) healthy normal controls who were not divers. Our results indicate that DCS increases the incidence of pathologic EEG recordings, whereas recompression treatment decreases them. The results of MRI do not verify evidence of increased numbers of CNS lesions in normal divers as compared to non-diving, healthy control subjects, whereas some of the divers treated for DCS in a pressure chamber had hyperintense lesions in brain white matter. None of them had any abnormalities in EEG, neurologic performance, or psychologic behavior. Both EEG and MRI are sensitive and non-specific methods for judging suspected evidence of brain lesions from diving or diving accidents. PMID- 10372425 TI - Simulated airplane flight increases plasma lactate in fetal rabbits. AB - We studied the effect of 9 h of simulated airplane cabin conditions at cruising altitude (8,000 feet; inspired oxygen equivalent to 15% O2 at sea level) on fetal plasma lactate in near-term pregnant rabbits. Controls (n = 19) spent 9 h at sea level (21% O2). Study group I (n = 21) experienced airplane cabin conditions. Study group II (n = 17) was studied at 8,000 feet with the inspired O2 concentration normalized to sea level. Study group III (n = 19) remained at sea level breathing 15% O2. Before ending each exposure, fetal blood sampling for lactate was performed under ultrasound guidance. Maternal lactates were obtained before and after sampling fetuses. Wilcoxon signed rank test, analysis of variance, and Bonferroni's method were used as appropriate. P < 0.05 denoted statistical significance. Study group I (altitude/hypoxia) had higher fetal lactates than controls (sea level/normoxia) and study group II (altitude/normoxia). Fetal lactates in study group I (altitude/hypoxia) were higher than in study group III (sea level/hypoxia). Maternal lactates were lower after fetal sampling. Fetal lactic acidemia was observed after 9 h of airplane cabin conditions. This was attributed to the combined effect of the lowered oxygen concentration and the decrease in atmospheric pressure. PMID- 10372426 TI - Glutathione in the cellular defense of human lung cells exposed to hyperoxia and high pressure. AB - Saturation diving involves exposure to elevated partial pressure of oxygen (Po2) and high pressure. The present work demonstrated that hyperoxic exposure for up to 72 h had significant effects on human lung fibroblasts. Forty to sixty kPa Po2 had severe acute toxic effects, and 60 kPa O2 reduced plating efficiency approximately 96% and completely inhibited cell proliferation. Long-term toxic effects were observed as a persistent reduction of cell growth rate after 24 h exposure to 60 kPa O2 in helium, suggesting genetic effects or induction of cellular senescence. No effect of high pressure per se was observed in this respect. Cellular glutathione was increased up to a plateau 40-50% above control level after an initial decrease, which may indicate toxic effects during the GSH depletion period. The glutathione egress increased even more than the intracellular level after exposure to these conditions. The effects on glutathione were growth state specific with the highest response in exponentially growing cells. Slight protective effects of high pressure were noted in a cell growth assay, correlating with a reduced response on the glutathione level. The results support previous studies indicating that hyperoxia is the main contributor to the adverse effects of exposure to high Po2 and high pressure and point to the involvement of glutathione in the cellular detoxification of reactive oxygen species under these conditions. PMID- 10372427 TI - Effect of hyperbaric conditions on plasma stress hormone levels and endothelin-1. AB - To study a generalized stress reaction as well as endothelin-1 concentrations during moderate hyperbaria and hyperbaric oxygen (HBO2), eight professional divers were exposed to air (O2 21%, AIR) and oxygen (O2 100%, HBO2) at 2.5 atm abs for 60 min in separate sessions. Plasma concentrations of epinephrine, norepinephrine, dihydroxyphenylglycol (metabolite of norepinephrine), cortisol, ADH, renin, aldosterone, pro-ANP, and endothelin-1 were analyzed before, during, and 20 min after the treatments. Endothelin-1 increased significantly (6% during HBO2 and 18% during AIR, and 30 and 34% after the treatments, respectively, P = 0.032). There was no statistically significant difference in the changes of mean norepinephrine and dihydroxyphenylglycol levels between the treatments, although both seemed to change slightly during the treatments, but not over the baseline (time effect P = 0.031 and P = 0.011, respectively). Cortisol levels decreased significantly (P = 0.001) during the treatments. No significant changes were found in other analyzed hormones. The authors concluded that a) HBO2, and hyperbaric air at 2.5 atm abs do not induce a generalized hormonal stress reaction, and b) endothelin-1 increases during HBO2 and hyperbaric air at 2.5 atm abs. PMID- 10372428 TI - Effect of caffeine consumption on tissue oxygen levels during hyperbaric oxygen treatment. AB - Ten men were exposed to hyperbaric oxygen (HBO2), and their tissue oxygen levels were monitored after they drank either placebo or caffeine beverages. Transcutaneous tissue oxygen (PtcO2) monitor measurements in a normobaric air environment were initially obtained from transducers on the subject's chest and foot. The subjects then consumed either the caffeine (3 mg.kg-1) or the placebo beverage, and after 20 min the subjects were pressurized in a hyperbaric chamber to 2.36 atm abs (1 atm = 101.3250 kPa). The test subjects began breathing 100% oxygen at 2.36 atm abs, 30 min after administration of the experimental beverage, and continued for 30 min, after which the final chest and foot PtcO2 measurements were recorded (1 h after ingestion of the test drink). Each subject underwent a second hyperbaric exposure during which the alternate drink was administered (either the placebo or the caffeine), and PtcO2 measurements were again obtained. The increase in right foot PtcO2 values during HBO2 exposure was significantly smaller after caffeine consumption than after placebo (P = 0.0018). PMID- 10372429 TI - Attenuation of brain hyperbaric oxygen toxicity by fasting is not related to ketosis. AB - The effect of 24 h of fasting and changes in blood glucose and beta hydroxybutyrate (BHB) level on latency to seizures in hyperbaric oxygen (HBO2) was studied. Conscious, unrestrained rats implanted with cortical electroencephalogram electrodes were exposed to 0.5 MPa (gauge pressure) O2 until seizures were observed. Fasting for 24 h significantly (P < 0.01) decreased blood glucose (from 8.6 +/- 0.9 in fed to 6.9 +/- 0.7 mM in the fasted group), increased blood BHB (0.07 +/- 0.02 mM to 0.38 +/- 0.10 mM, respectively), and prolonged the latency to seizures compared with normally fed animals (21.0 +/- 9.8 vs. 34.6 +/- 17.7 min, P < 0.05). Injection of the ketone precursor 1,3 butanediol (BD) to the fed animals increased blood BHB level to 0.72 +/- 0.32; however, seizure latency remained the same as in fed animals. Restoration of blood glucose in fasted animals to the same level as in the fed group did not reverse the protection achieved by fast; instead it increased the latency to seizures. The results indicate that the protection against HBO2 seizures by fasting in short starvation is not related to the increase in circulating ketone bodies or decrease in blood glucose. PMID- 10372430 TI - Effect of the anti-motion-sickness medication cinnarizine on central nervous system oxygen toxicity. AB - Severe seasickness could pose a serious problem in diving, and anti-seasickness medication should therefore be prescribed for the seasickness-susceptible diver. Cinnarizine may be used as a medication if it does not increase the risk of central nervous system (CNS) oxygen toxicity when diving with closed-circuit oxygen or O2-enriched gas mixtures. Twenty-six male, white Sprague-Dawley rats were exposed to high O2 pressures (507 and 608 kPa) before and after cinnarizine ingestion (3.3 mg.kg-1), until the appearance of the first electrical discharge (FED) in the electroencephalogram (EEG) which precedes the clinical convulsions. Each rat was tested on five exposure protocols (control and cinnarizine at 507 kPa O2, control, cinnarizine, and 15 h starvation as a control for cinnarizine at 608 kPa O2) at intervals of at least 2 days or until the EEG connector became detached (a mean of 3.1 exposures per rat). Latency to the FED increased after cinnarizine ingestion in 16 of the 17 pairs of measurements at 507 kPa O2 (by more than 61%, P < 0.002) and in 17 of the 19 pairs of measurements at 608 kPa O2 (by 36%, P < 0.002). There was no significant effect of 15 h starvation. Cinnarizine can be further considered for use in seasickness-susceptible divers as it does not increase the risk of CNS O2 toxicity. PMID- 10372431 TI - Physiologic and biochemical monitoring during hyperbaric oxygenation: a review. AB - Hyperbaric oxygenation (HBO2) is an important treatment given to various groups of patients exposed to pathologic situations (i.e., carbon monoxide exposure). Since many hyperbaric patients are critically ill and are being treated for life threatening disorders, it is necessary to monitor various physiologic and biochemical parameters. This is a review of 193 publications covering a wide range of monitored parameters representing metabolic, hemodynamic, respiratory, electrical, and biochemical activities. The significance of monitoring the physiologic, medical, and specific oxygen toxicity effects during HBO2 exposure (MHBO2) is described and emphasized. Further development of new monitoring devices and technologies will enable the improvement of patient management during HBO2 treatment given under various medical conditions. This review also presents new ideas about possible future monitoring of brain function under HBO2 conditions in experimental animals as well as under clinical conditions. PMID- 10372432 TI - [Anesthetic problems during long interventions in mouth, jaw and facial surgery]. AB - Long-lasting and extended maxillofacial surgery with operating times of more than ten hours are now routinely performed with an acceptable risk for the patient. Careful preoperative evaluation of the patient and interdisciplinary planning are essential for successful surgery. The close proximity of the surgical field and the airways as well as a high incidence of difficult intubation in these patients require special attention from the anaesthesiologist. Other perioperative focuses of anaesthesiological concern in long-lasting maxillofacial surgery are positioning of the patient, intra- and postoperative airway management, intraoperative monitoring, thermoregulation, fluid replacement and transfusion therapy. PMID- 10372433 TI - [Intraoperative changes in arterial end tidal CO2 partial pressure difference in interventions with constant ventilation-perfusion ratio]. AB - During general anaesthesia, the endtidal CO2 pressure serves as an estimate of the arterial CO2 pressure to regulate the ventilator setting. Important arterial to end-tidal carbon dioxide tension differences (P(a-et)CO2) have been observed among patients undergoing procedures which have substantial impact on the ventilation-perfusion ratio (V/Q). Data on the P(a-et)CO2 for procedures in which the V/Q-ratio remains constant are lacking. Repeated measurements of P(a-et)CO2 in twelve patients with chronic obstructive lung disease (COLD) and nine pulmonary healthy patients undergoing jaw surgery were performed. The P(a-et)CO2 in the pulmonary healthy subjects (5.96 +/- 1.68 mmHg) was lower than in the COLD patients (9.05 +/- 3.49 mmHg) (p < 0.01). A clinically significant P(a-et)CO2 > or = 8 mmHg was observed in 52% of the measurements in patients with COLD compared with 11% in the pulmonary healthy subjects (p < 0.01). Both patient groups showed only minimal intraoperative changes of P(a-et)CO2. The deviation of all subsequent P(a-et)CO2 values from the initial P(a-et)CO2 was 2.17 +/- 1.52 mmHg in the pulmonary healthy patients and 2.02 +/- 1.49 mmHg in the patients with COLD (p = 0.76). Intraoperative changes of the P(a-et)CO2 are small during procedures with no major alterations of the V/Q ratio. For these procedures an initial measurement of the P(a-et)CO2 in patients with lung disease should be sufficient. In pulmonary healthy subjects the P(a-et)CO2 seems to be negligible. PMID- 10372434 TI - [Minimal flow anesthesia in newborn infants--advantages and risks]. AB - The long predominance of the semi-open anaesthetic system in paediatric anaesthesia has been ended by the introduction of circle systems by Altemeyer. Narcoses in newborn infants, however, are usually performed with a circle system and a fresh gas flow (FGF) that greatly exceeds the ventilation volume per minute required. This prevents a desirable degree of gas climatisation. A reduction of fresh gas flow for anaesthesia in neonates makes high demands on the anaesthesia ventilators. The safety and precision of present anaesthesia ventilators with different principles of function and construction were studied by means of a lung model reducing the FGF from 4.0 l/min to 0.5 l/min. In order to clarify the importance of a reduction of the FGF for the climatisation of anaesthetic gases and heat regulation in neonates we measured the temperatures of the respiratory gas at the tip of the tube and the body temperatures with a temperature sound. We compared 42 newborn patients anaesthetized with either high gas flow (3.0 l/min) or minimal gas flow. Our results showed that ventilators suitable for safely reducing FGF in neonates are available. Not every ventilator, however, offers the degree of precision required. Depending on FGF heat regulation in newborn infants differed significantly. Using high flow ventilation respiratory gas and rectal temperatures declined continuously. When FGF was reduced there was a significant increase of temperature parameters after 25 min (gas) and 35 min (body). Body temperature came back to normal values or stayed normal. Artificial ventilation of neonates in anaesthesia lasting more than 50 minutes should routinely be performed with minimal FGF in order to ensure normothermia. PMID- 10372435 TI - [Ventricular rupture after blunt thoracic trauma]. AB - In a collision with a motor-car, a pedestrian suffered multiple injuries and a blunt trauma to the thorax. Immediately after the accident, the patient was haemodynamically instable and needed resuscitation several times, without lasting success. The coroner's office found that cardiac tamponade from a ruptured right ventricle was the cause of death. The incidence of ventricular rupture due to blunt trauma in motor-car accidents is about 10 to 15%. Since definite treatment is not possible at the site of the accident, the patient must be taken immediately to a cardio-surgical hospital after initial stabilization. Unfortunately, preclinical diagnosis of ventricular rupture is difficult. In this context, the increasing availability in ambulances of a 12-channel ECG, a highly sensitive diagnostic tool, represents significant progress. Cases like the one described above should be discussed at mortality conferences of pathologists, coroners and emergency physicians to increase awareness of this problem. Only if the possibility of cardiac rupture is considered and ruled out early in cases of massive multiple injuries with haemodynamic instabilities, will decrease the apallingly high lethality figures. PMID- 10372436 TI - [Effect of local anesthetics on hemodynamic effects during Mayfield skull clamp fixation in neurosurgery using total intravenous anesthesia]. AB - For neurosurgical procedures, the association between insertion of the Mayfield skull clamp and haemodynamic changes is generally recognized. We investigated the protective effect of two local anaesthetic substances (lidocaine and bupivacaine) under the conditions of total intravenous anaesthesia (TIVA) with propofol and alfentanil. Forty-two patients undergoing an elective craniotomy (tumor resection) were included in the study and randomly divided into three groups. All patients were given a total intravenous anaesthesia with propofol and aflfentanil. After induction, the skin areas for the pin were infiltrated with 0.9% sodium chloride (n = 14, control group 1), 1% lidocain (n = 14, group 2) or 0.5% bupivacaine (n = 14, group 3). After an interval of 1 to 2 minutes the pins were inserted. The intra-arterial line was inserted before induction. The haemodynamic parameters heart rate (HR), systolic arterial pressure (SAP), diastolic arterial pressure (DAP) and mean arterial pressure (MAP) were monitored continuously. The haemodynamic parameters were recorded at four set times: (1) after induction of anaesthesia, (2) at the onset of the local anaesthesia, (3) at the insertion of the pin-holder, (4) five minutes after insertion. Insertion of the pins led to a significant increase in HR, SAP, MAP and DAP in the control group. These haemodynamic changes can be reduced by local infiltration with lidocaine or bupivacaine. The effect of both substances was the same in our study. Our results suggest that a significant reduction of the haemodynamic effects caused by insertion of the Mayfield skull clamp can be achieved by the use of local anaesthesia. Total intravenous anaesthesia alone with propofol and alfentanil cannot protect against these haemodynamic stimuli. PMID- 10372437 TI - Evidence-based medicine: a new paradigm? PMID- 10372438 TI - Evidence-based medicine. PMID- 10372439 TI - Specific and reversible inhibition of Entamoeba histolytica cysteine-proteinase activities by Zn2+: implications for adhesion and cell damage. AB - BACKGROUND: Cysteine-proteinases are thought to play an important role in E. histolytica pathogenicity. Although effective blockage of proteolytic activities can be obtained with several known inhibitors, the high cellular toxicity of most of the inhibitors precludes experimentation with live cells or animal models. Specific cysteine-proteinase inhibitors that could be utilized in studies of virulence are of great need in the field of amebiasis. METHODS: Cysteine proteinase activities were determined in trophozoite lysates by azocasein degradation and after PAGE and gelatin zymograms. Inhibition of the activities was assessed in the presence of 0.01-2.5 mM concentrations of divalent cations of the IIB and VIII series such as Zn, Cd, Hg, Ni, and Co. Reversibility was induced with 25 mM L-cysteine or 50 mM L-histidine and by metal chelation with 5 mM phenantroline. The inhibitory effect of ZnCl2 was tested with live cells in fibronectin-binding and cytotoxicity assays. RESULTS: ZnCl2 specifically inhibited cysteine-proteinase activities in trophozoite lysates in a concentration-dependent manner. Additionally, 1.0-2.5 mM ZnCl2 blocked proteolysis in more than 70%. This inhibition was completely reverted by L cysteine, L-histidine, or phenantroline. Similar results were obtained by analyzing individual cysteine-proteinase activities separated in gelatin zymograms. At these concentrations, ZnCl2 significantly interfered with trophozoite adhesion, thus making amebas deficient in substrate degradation and cell damage. CONCLUSIONS: ZnCl2 is an effective inhibitor of amebic cysteine proteinases. Its low toxicity at relatively high concentrations, high solubility, and low cost make it adequate for live cell experimentation and animal models of amebic virulence. PMID- 10372440 TI - Patterns of adrenoceptor change during liver regeneration of the Wistar Kyoto rat: a binding study. AB - BACKGROUND: Adrenoceptors have been involved in the regulation of hepatocyte proliferation after partial hepatectomy, as well as in primary culture. This report characterizes alpha 1- and beta-adrenoceptor change during the time-course of liver regeneration in adult Wistar Kyoto rats. METHODS: Saturation binding assays with [3H]prazosin or [3H]dihydroalprenolol (for alpha 1- and beta adrenoceptors, respectively) were done in liver plasma membranes from 6-month-old rats subjected to 70% hepatectomy followed by hepatic regeneration. RESULTS: [3H]Prazosin and [3H]dihydroalprenolol binding gave control Bmax values of 101 +/ 10 and 12 +/- 1 fmol/mg protein and Kd of 0.50 +/- 0.10 and 4.1 +/- 0.4 nM for alpha 1- and beta-adrenoceptors, respectively. alpha 1-Adrenoceptor number and Kd increased at 24 and 48 h and returned to control values at 72 and 96 h after surgery, whereas beta-adrenoceptors augmented at 48 and 72 h, with a Kd change at 24 and 48 h posthepatectomy. CONCLUSIONS: These results suggest that dual control of alpha 1- and beta-adrenoceptor membrane expression could be involved in different steps during hepatocyte proliferation, and that Wistar Kyoto rats have a different adrenoceptor pattern expression from other rat strains. PMID- 10372441 TI - Superoxide dismutase activity of the salicylate-iron complex. AB - BACKGROUND: Scavenging of superoxide radical by salicylate-iron complex was studied to determine whether or not the salicylate-iron complex was able to catalyze the dismutation of superoxide radicals, the result perhaps yielding an explanation of the antioxidant and anti-inflammatory properties of the drug. METHODS: The scavenging was studied with an assay that generates O2.- without the intervention of metal ions. RESULTS: Results indicated that, in the presence of iron, salicylate was able to bring about the catalytic dismutation of the superoxide radical. The rate of superoxide removal was dependent on both the concentration of iron and the salicylate:iron molar ratio. CONCLUSIONS: These results may help to explain the interaction of nonsteroidal anti-inflammatory drugs with free radicals and the anti-inflammatory properties of these agents, inasmuch as accumulating evidence indicates that much of the injury observed during inflammatory disorders may be mediated by oxidative stress frequently induced by iron-dependent reactions. PMID- 10372442 TI - Surface redistribution of interferon gamma-receptor and its colocalization with the actin cytoskeleton. AB - BACKGROUND: Specific antibodies for human IFN gamma-R1 were used to examine its mobilization in Colo 205 cells. METHODS: We report here that antibody-IFN gamma R1 complex induced capping and actin colocalization. Pretreatment with cytochalasin D abolished this capping. To define the role of the IFN gamma-R1 in the possible interaction with actin, transfected murine fibroblasts cell line with human cDNA IFN gamma-R1 were used. RESULTS: Only those cells expressing the full receptor and cultured in suspension polarized the receptor and this colocalized with actin filaments. Nevertheless, cells truncated in their intracellular domain displayed no capping and actin remained unaltered either in suspension or in monolayer culture conditions. A mutant bearing an IFN gamma-R1 with substitutions in positions 270-271 of the intracellular domain redistributed both IFN gamma-R1 and actin as micropatches instead of capping. Mutation in 256 303 residues resulted in IFN gamma-R1 microaggregates but actin remained unchanged. CONCLUSIONS: These experimental models allowed us to highlight an apparent receptor-microfilament association through the intracellular domain of IFN gamma-R1, and to specifically locate it within the intracellular region 256 303 that has been identified as relevant for ligand-receptor internalization and biological function. PMID- 10372443 TI - Congo red effect on cyst viability and cell wall structure of encysting. Entamoeba invadens. AB - BACKGROUND: The cell wall of Entamoeba invadens cysts is composed of chitin microfibrils as the main structural component. It has been demonstrated in yeast that the chitin cell wall assembly is altered by dyes such as Congo red (CR) and Calcofluor. METHODS: The purpose of this work was to study the cell wall assembly under the effect of CR dye on encysting E. invadens by means of light and electron microscopy, after the amebas were subjected to the effect of 100-2,000 micrograms CR/mL. Experiments were performed either in BI-S-33 or in mLG media. RESULTS: Trophozoite growth was not inhibited by 100-1,000 micrograms/mL CR after 8 days of incubation in BI-S-33 medium. However, low levels of growth were observed with 2,000 micrograms/mL of dye. No significant differences in morphologically viable (hyaline) cyst production occurred after 24-48 h, when 100 micrograms CR/mL was used, while the highest concentration of CR (2,000 micrograms/mL) resulted in a significant decrease of hyaline cyst yield; dead cysts prevailed in cultures, particularly at 72 h of CR treatment. Differentiation of amebas incubated in the presence of 500-2,000 micrograms/mL CR produced abnormal chitin deposits, rendering irregularly thick or double cell walls, as shown by transmission and scanning electron microscopy. Cyst cultures obtained under 100 micrograms/mL CR produced as many trophozoites as did the control when they were incubated in BI-S-33, but only low numbers of trophozoites were found in culture cysts obtained under higher CR doses. CONCLUSION: Our results suggest that CR affects E. invadens encystment, alters the cell wall formation, and also affects the cyst viability. PMID- 10372444 TI - Heparin and low molecular weight heparin decrease nitric oxide production by human polymorphonuclear cells. AB - BACKGROUND: Heparin and heparin derivatives with low anticoagulant activity exhibit a wide spectrum of biological functions affecting adhesion, activation and trafficking of leukocytes. METHODS: We investigated the in vitro effect of heparin and a low molecular weight heparin derivative (LMWH) on nitric oxide (NO) production by human polymorphonuclear leukocytes (PMN). RESULTS: N-formyl methionyl-leucyl-phenylalanine (fMLP)-stimulated NO production was significantly decreased by heparin at doses of 0.5 and 5 micrograms/mL, while LMWH was only effective at doses of 50 and 200 micrograms/mL by means of a mechanism not related to NO synthase (NOS) activity. CONCLUSIONS: These results support the hypothesis that heparin and LMWH derivatives may offer therapeutic benefit for inflammatory diseases where NO plays a protagonic role. PMID- 10372445 TI - Microtia: a clinical and genetic study at the National Institute of Pediatrics in Mexico City. AB - BACKGROUND: Microtia is a malformation of the ear with extreme variability of expression. It is generally seen as an isolated malformation. However, some authors consider it to be a minimal manifestation of the oculo-auriculo-vertebral spectrum (OAVS), where, in addition, there are facial, vertebral, and renal abnormalities, among others. METHODS: A total of 145 pediatric patients with unilateral or bilateral microtia not considered as part of a syndrome were studied. All patients were subjected to an intentional clinical examination, a familial history, and radiographic imaging studies for ruling out associated malformations. Patients were classified into two groups: group 1 (60%), with isolated microtia; and group 2 (40%), considered as OAVS, with microtia associated with hemifacial skeletal microsomia, vertebral and/or renal malformations. RESULTS: No significant differences were found between the groups when the following variables were compared: gender; presence of unilateral or bilateral microtia; atretic external auditory canal; presence of preauricular tags; hearing loss of any type, and affection of the seventh cranial nerve, as well as associated malformations of other organs or systems. There were significant differences in relation to the presence of soft-tissue hemifacial microsomia, more frequently seen in patients with OAVS, because the majority of these patients had bone microsomia. Over 66% of the cases were sporadic and the rest were familiar. In 28.3% of the cases, the history suggested an autosomal dominant inheritance pattern, and in 5.5%, an autosomal-recessive inheritance pattern, although in some familial cases, multifactorial inheritance could not be ignored. Some members in several families had isolated microtia, and others had mild characteristic manifestations of OAVS. CONCLUSIONS: Our results support the hypothesis that isolated microtia is a minimal expression of OAVS. Therefore, it is recommended that patients with microtia be subjected to intentional studies that search for malformations and physical examinations of first-degree relatives for adequate genetic counseling and management. PMID- 10372446 TI - Neurotoxicity of dextrorphan. AB - BACKGROUND: The noncompetitive NMDA antagonists phencyclidine (PCP) and dizocilpine (MK-801) have been considered for use as neuroprotective therapeutic agents, although both produce injury in neurons of cingulate and retrosplenial cortices in rodents. The low-affinity, noncompetitive NMDA antagonist dextrorphan has been considered for use as a neuroprotective therapeutic drug. The aim of the present work was to evaluate the neurotoxicity of dextrorphan. METHODS: Sprague Dawley male rats were used and injected with either saline or dextrorphan (30 mg/kg i.p.). The animals were sacrificed 30 min later, and the brain was examined for histopathological changes. RESULTS: After systemic administration of the drug, hyperchromatic and shrunken nuclei with chromatin condensation and disruption were observed. Also, granular and vacuolated cytoplasm was apparent in pyramidal neurons in the retrosplenial (posterior cingulate) cortex. Status spongiosus (spongy degeneration) of the neuropil was also detected. CONCLUSIONS: Morphological changes are similar to those described previously, which are induced by high-affinity, noncompetitive NMDA antagonists, such as MK-801. PMID- 10372447 TI - Security and maximal tolerated doses of fluvastatin in pediatric cancer patients. AB - BACKGROUND: The role of cholesterol in neoplasic cell growth and its inhibition by drugs has recently been studied. Cholesterol biosynthesis inhibitors have been used as adjuvants in the treatment of cancer and possibly as prophylactic in carcinogenesis. OBJECTIVE: The objective of the study was to determine the maximal tolerated doses (MTD) and toxic effects of fluvastatin in pediatric cancer patients. METHODS: This study was carried out in a third level Social Security Hospital in Mexico City. We included pediatric patients from April 1996 to May 1997. All were terminal cancer patients who did not respond to conventional therapies. Fluvastatin was given p.o. at doses of 2 mg/kg/day for 14 days every 4 weeks in three patients. Subsequent cohorts of three patients each had increments of 2 mg/kg/day of the drug until maximal tolerated doses were found. Toxic effects of the drug were evaluated by physical exploration, laboratory assays and a questionnaire given to each patient. RESULTS: Twelve patients were included. Diagnoses included two osteosarcomas, eight central nervous system tumors, one lung tumor, and one Ewing's sarcoma. Ten patients died within 1 to 18 months. Two are alive 22 months after inclusion into the study, both with anaplasic astrocytoma. A total of 27 courses were administered. The MTD was 8 mg/kg/day. Toxic effects were insomnia, nausea, vomiting, abdominal distention and myalgias. Toxicity was dose-dependent. Laboratory assays demonstrated no significant changes during treatment. CONCLUSIONS: Fluvastatin can be safely used at doses of 8 mg/kg/day in pediatric patients with cancer. This dose should be used in additional trials. PMID- 10372449 TI - Tumor necrosis factor in peritoneal fluid from asymptomatic infertile women. AB - BACKGROUND: Tumor necrosis factor-alpha (TNF-alpha) is a cytokine that can be found in the peritoneal fluid (PF) of patients with endometriosis and pelvic inflammatory disease (PID) as a response to inflammatory disorders and infections. The cytotoxic effect of this cytokine could be a factor participating in the pathology of various gynecological diseases, and could also be accountable for the high immunological response and damage to the tubal epithelium. The objective of this study was to establish the presence of TNF-alpha in asymptomatic infertility and its association with various isolated bacteria. METHODS: Ten milliliters of PF were collected from each of 73 patients by means of laparoscopy and cultured in synthetic medium and McCoy cells for the isolation of aerobic and anaerobic bacteria, as well as for Chlamydia trachomatis. The activity of TNF-alpha was determined by means of a bioassay using L-929 cells. RESULTS: Forty-three percent of the PFs showed positive TNF-alpha activity, while the laparoscopic evaluation showed that 32 patients had Fallopian tube occlusion (FTO), 7 had endometriosis, 30 had PID, and 4 had myomas and adhesions. TNF-alpha activity was found to be high in FTO patients (p < 0.05). Positive cultures were found in 50.7% of patients; of these, 31.5% had PID (p < 0.05), and only 20.5% of positive cultures were TNF-alpha positive. Chlamydia trachomatis (16%) was the most frequently isolated bacteria in these patients. CONCLUSIONS: The detection of TNF-alpha could be useful in the diagnosis of active infectious and inflammatory diseases in asymptomatic infertile patients. PMID- 10372448 TI - Effect of superoxide dismutase from bovine erythrocytes on different activity parameters in adjuvant-induced arthritis. AB - BACKGROUND: The purpose of this work was to evaluate the effect of superoxide dismutase (SOD) on primary swelling, lipoperoxidation, body thymus, and spleen weight in the adjuvant-induced arthritis (AIA) model in rats. METHODS: Orally and intraperitoneally administered SOD (100 U/kg) from bovine erythrocytes, as well as naproxen (40 mg/kg) and dexamethasone (25 mg/kg), were evaluated against placebo. RESULTS: Primary edema was not decreased by SOD; in contrast, naproxen and dexamethasone showed good anti-inflammatory activity. Lipoperoxidation increased 1.8, 2.5, and 2.8 times with intraperitoneal SOD, naproxen, and dexamethasone administration, respectively, while oral SOD decreased lipoperoxidation levels to approximately one-half of that found in the control group. Body weight increased with SOD but decreased with dexamethasone. Naproxen did not change the animal weight. Thymus weight remained unchanged with SOD and naproxen, while it decreased with dexamethasone. Spleen weight remained the same with SOD, but increased with naproxen and decreased with dexamethasone. No side effects were observed in the SOD group, whereas 20% of the rats in the naproxen group died of gastrointestinal hemorrhage, and 50% of the rats in the dexamethasone group, of pulmonary infection. CONCLUSIONS: In conclusion, SOD showed no anti-inflammatory activity but decreased lipoperoxidation when administered orally. No deleterious effects in primary and secondary immunologic organs were observed with this agent. PMID- 10372451 TI - Malnutrition in childhood lymphoblastic leukemia: a predictor of early mortality during the induction-to-remission phase of the treatment. AB - BACKGROUND: Previous reports have shown that undernourished children with acute lymphoblastic leukemia (ALL) have a poorer long-term survival as compared with children with normal nourishment status. It has been shown that both the relapse and mortality rates of undernourished children with ALL are higher during the continuation phase of the chemotherapy and are apparently related to a poor tolerance of ablative chemotherapy. No previous articles have analyzed the early mortality rate of these patients. METHODS: We carried out a case-control study, and have studied the effect of severe malnutrition on the mortality of 17 children with ALL during the initial induction-to-remission phase of the treatment. These 17 cases were compared with 76 controls who had survived at least the phases of induction and consolidation. RESULTS: It was found that the chance of dying during the initial phase of the treatment was 2.6 times higher (confidence interval 95%: 0.55-11.89) in undernourished children with ALL than in those children with normal nourishment status. The risk of death increased with the severity of undernourishment (p = 0.04). CONCLUSIONS: These data confirm the prognostic value of malnutrition in children with ALL and suggest that undernourishment may also influence early mortality during the induction-to remission phase of the treatment. PMID- 10372450 TI - Discrimination between epidemiological cycles of rabies in Mexico. AB - BACKGROUND: The design of efficient rabies control programs within a geographic area requires an appropriate knowledge of the local epidemiological cycles. In Latin America, there is a geographical overlap of the two main epidemiological cycles: (a) the terrestrial cycle, where the dog is the main terrestrial vector and the principal cause of human transmission; and (b) the aerial cycle, in which the vampire bat Desmodus rotundus is representative in Mexico. This bat is the major sylvatic rabies vector transmitting rabies to cattle. The purpose of this study was to distinguish between the epidemiological cycles of rabies virus (aerial and terrestrial) circulating in Mexico, using restriction fragment length polymorphism (RFLP). METHODS: Thirty positive rabies isolates were obtained from different species (including humans, domestic, and wildlife animals) and geographical regions. The methodology included the extraction of RNA, and synthesis of cDNA, PCR, and RFLP using four restriction endonucleases. To determine the aerial cycle, BsaW I and BsrG I were utilized, and for terrestrial cycle, BamH I and Stu I. Most of the samples belonged to the aerial and terrestrial cycles, except for two skunk isolates from Northwestern Mexico, which were not cut by any of the enzymes. RESULTS: Three different migration patterns were detected: (a) the first was observed in six amplicons, which were cut by BsaW I and BsrG I (aerial cycle); (b) 19 amplified samples were digested with BamH I and Stu I enzymes (terrestrial cycle); and (c) two skunk isolates from Northwest Mexico, were not cut by any of the enzymes utilized in the experiments (hypervariable cycle). CONCLUSIONS: This concludes that RFLP can be used for the classification of rabies field samples in epidemiological studies. Moreover, it has demonstrated its usefulness, not only for differentiating between the main epidemiological rabies cycles present in Mexico, but also to detect new cycles in wildlife species. PMID- 10372452 TI - Possible relationship between neurocysticercosis and hematological malignancies. AB - BACKGROUND: Previous studies have shown an increased frequency of chromosomal abnormalities in lymphocytes from animals and humans with cysticercosis. Some reports have suggested an association between cancer and cysticercosis. The aim of this study was to investigate the possible association between neurocysticercosis and cancer. METHODS: We designed a mortality rate study from the autopsy files of the Department of Pathology at the General Hospital of Mexico. A total of 1,271 autopsy files were reviewed. All files in which a malignant neoplasia was found during autopsy were selected as cases. Autopsies in which no malignant disease was found were used as controls. The odds ratio was determined between the frequency of neurocysticercosis in patients with any malignant neoplasia and that of the controls. RESULTS: Neurocysticercosis was more frequent in cases with malignant hematological diseases (MHD) than in controls (p = 0.01). The odds ratio for this association was 3.54, with 95% confidence interval from 1.17-9.79. CONCLUSIONS: Most human cancers arise from the interaction of a multiplicity of factors, including xenobiotics and endogenous constituents. Therefore, while it will be difficult to demonstrate that neurocysticercosis is a causal agent of malignant hematological diseases (MHD), it should be considered as a potential risk factor for cancer induction in countries where cysticercosis remains a public health problem. PMID- 10372453 TI - [Caring for menopause in primary care]. PMID- 10372454 TI - [Tobacco consumption in school-children from the autonomous community of Extremadura]. AB - OBJECTIVES: To find the number of smokers among school-children in Extremadura in the eighth year of EGB/second of ESO, the relationship of their environment with the acquisition of the habit and their attitudes towards tobacco. DESIGN: Crossover study of a representative sample of the students doing these courses. SETTING: All the schools in our autonomous community. PARTICIPANTS: 1062 students from various schools. MEASUREMENTS: We used a self-administered questionnaire with questions on the tobacco consumption of the interviewees, their parents, siblings, friends and teachers. The study included other variables, such as age, sex, alcohol consumption, parents' educational qualifications and other questions pertinent to attitudes to tobacco. RESULTS: The age of those surveyed was 13-14 (54% boys), of whom 18.27% were habitual smokers (95% CI, 15.95-20.59). Tobacco consumption was higher in boys (20.48% against 15.24% in girls; p = 0.05). The habit is clearly higher in friends, siblings, fathers and mothers of smokers (p < 0.001; < 0.001; < 0.001, and < 0.01, respectively), but not in teachers. Parents of non-smokers had higher educational qualifications. Smokers showed favourable attitudes towards consumption and a positive association with alcohol intake. CONCLUSIONS: Given their age, the number of smokers is high and is associated with the presence of the habit in their most immediate social environment. Attitudes in our school-children which favour consumption justify the introduction of prevention programmes. PMID- 10372455 TI - [White coat and non-dipper hypertension in patients recently diagnosed with mild hypertension]. AB - OBJECTIVE: To calculate the prevalence of white coat hypertension (WCH) in patients recently diagnosed with light hypertension. To compare their demographic features, cardiovascular risk factors, and the level of early organic effect of WCH versus sustained hypertension, and dippers versus non-dippers. DESIGN: Descriptive, crossover study. SETTING: Five urban health centres. PATIENTS: 238 people between 18 and 65 were chosen. After screening, they were diagnosed with light (1993 WHO criteria) or essential hypertension. MEASUREMENTS AND MAIN RESULTS: Patients received: 24-hour out-patient control of blood pressure (BP), analysis, back of eye and electrocardiogram. WCH was defined as mean daily BP < 139/88 mmHg and mean night-time BP < 123/74 mmHg. Non-dippers were those patients whose mean night-time BP went down from the daytime BP by less than 10%. 39.5% had WCH (33.3-45.7). This was associated with women (49.5%), with lower casual systolic and diastolic BP and with isolated systolic hypertension, p < 0.05. Risk of WCH was 2.14 times greater in women (95% CI, 1.24-3.70). There were no significant differences in the cardiovascular risk or morbidity profile between WCH and sustained HT, or between dippers and non-dippers. CONCLUSIONS: WCH is common in patients recently diagnosed with both light and essential hypertension. This makes us think that the use of primary care out-patient monitoring of BP could be efficient in this type of patient. The absence of significant differences between WCH and sustained hypertension, or between dippers and non dippers, may be due to their hypertension being recent. PMID- 10372456 TI - [Impact of various strategies on the rates of flu vaccination in the elderly]. AB - OBJECTIVE: To calculate the impact of different interventions on the rates of anti-flu vaccination in people aged 65 and over. DESIGN: Ecological study with analytical components. SETTING: Health centres and clinics in two health areas in the Community of Valencia. PATIENTS AND OTHER PARTICIPANTS: Care units, characterised by their coverage of stable nuclei of 500 or more people, total population of 415,172 inhabitants. INTERVENTIONS: Use of the communication media, personal invitations by post and telephone; follow-up of activity, discussion of results, external evaluations; registration of people to be vaccinated and prior order to nursing staff to vaccinate. MEASUREMENTS AND MAIN RESULTS: The coverage in quartiles of the previous season and the human resources per thousand inhabitants were the structural factors which best explained the vaccination rates. In the multivariate analysis, invitation by letter had relative risk (RR) of 1.062 (95% CI, 1.036-1.088); telephone call, RR 1.075 (95% CI, 1.021-1.132); discussion of external evaluations, RR 1.046 (95% CI, 1.024-1.068); and the order of vaccination to nursing staff, RR 1.056 (95% CI, 1.025-1.088). All these were associated with greater coverage. The absolute difference in coverage achieved between the care units which invited people to be vaccinated by letter and phone, at which nurses vaccinated directly, and which discussed external evaluations, and the units which did not perform these above activities, was 14 points (95% CI, 13.3-14.5), independently of the other factors. CONCLUSIONS: The previous year's coverage, human resources, invitation by letter or phone, the order to nursing to vaccinate and the discussion of external evaluations, were all associated, independently, with higher rates of vaccination. PMID- 10372457 TI - [Primary care population screening for growth]. AB - OBJECTIVE: To find the effectiveness of growth screening of 6 and 7-year olds in primary care and the characteristics of slow growth found. DESIGN: Crossover, observational study. PATIENTS: 6 to 7-year old population attended at 8 health centres in the province of Barcelona. INTERVENTIONS: Exploration of height and weight. The population with height=percentile 3 was referred to the paediatric endocrinologist for diagnosis. RESULTS: 2306 children (45% of the population attended) were screened. 73 had low height (3.5%). Of these, 8.2% did not attend the appointment with the endocrinologist; 5.5% had been wrongly measured, and for 19% no definite diagnosis could be reached. 49 cases were classified as cases of slow growth: 11 organic and 38 non-organic. The effectiveness of the screening was one unidentified case of slow growth in every 80 children screened. CONCLUSIONS: It would seem advisable to maintain the practice of screening for slow growth in primary care. To improve its efficiency, it is proposed to refer to the specialist only the population with their height under the percentile 0.4. The paediatrics team should deal with non-organic growth disorders. PMID- 10372458 TI - [Potential impact of measures to minimize drug costs in Aragon's primary care system]. AB - OBJECTIVE: To analyse the potential saving in pharmaceutical expenditure which the use of drug products of the same composition and lower price would suppose, by three high-consumption therapeutic sub-groups. DESIGN: Retrospective observation study. SETTING: Primary care. PARTICIPANTS: All the medical prescriptions of the doctors in the 121 primary care health districts in the three provinces of the Autonomous Community of Aragon during 1997, divided into the sub-groups: peptic ulcer (AO2B), lipid-lowering (B04A) and hypotensor drugs (C02). MEASUREMENTS AND MAIN RESULTS: Consumption, cost of treatment per day and potential saving of eight active principles belonging to these 3 sub-groups were calculated. Potential overall saving reached the figure of 972 million pesetas in Aragon in 1997. CONCLUSIONS: The alternative of prescribing drugs with an identical composition and lower price offers great potential savings in pharmaceutical expenditure, and may be a useful measure to improve the efficiency of health resources, and in particular those devoted to drug prescription. PMID- 10372459 TI - [Trends in smoking among school children: Barcelona, 1987-1996]. AB - OBJECTIVE: Despite the growth of aids and of other causes of premature death, smoking is the main single cause of avoidable mortality in Spain. Given the addictive nature of tobacco and the difficulties experienced by many smokers to quit, the primary prevention of smoking is crucial. In recent years several initiatives have been developed to this end. The objective of this paper is to monitor smoking among schoolchildren by repeated surveys over a decade, to ascertain the impact of current interventions and to reorient them according to evidence. DESIGN: Cross sectional surveys over a decade, from 1987 to 1996, while several smoking prevention efforts were developed. Survey instruments were similar self administered and anonymous questionnaires. SITE: Schools in Barcelona (Catalonia, Spain). PARTICIPANTS: Around 1000 participants in each survey were selected through representative samples of 8th grade students (predominantly 13-14 years old), stratified by school type and size in each round. MAIN RESULTS: Ever smoking declined, specially for boys (global decline 12.4%, p = 0.05). There is a clear and steady decline, statistically significant, in the proportion smoking in the month prior to the survey (global decline 50.5%, p < 0.0001). The proportion of regular smokers declines from 1987 to 1994, and then is stabilized (global decline 23%, p < 0.05). The proportion of daily smokers fluctuates, but globally it is the lowest of the decade in 1996. No changes are visible in the proportion who declare buying tobacco for personal use. DISCUSSION: From 1987 to 1996 several smoking indicators show a decline among 8th grade schoolchildren in Barcelona, Spain. The pattern suggests a decline in global smoking experimentation and in the intensity of experimentation, with a small reduction in the prevalence of regular use, while the small proportion of daily smokers at this young age does not change. PMID- 10372460 TI - [The frequency of illnesses attended and its relationship with the maintenance of the family doctor's skill]. AB - OBJECTIVE: To describe the incidence of health problems dealt with less often in primary care medical consultations, and to discuss its relationship with the maintenance of professional skill, with in-work training and case-load planning. DESIGN: A prospective observational study based on a year's recording. SETTING: The clinics of 44 primary care doctors from 10 autonomous communities. PATIENTS: 418,98 people were attended. INTERVENTIONS: The unit of analysis was the care episode. The incidence per 1000 people attended, in total and by demographic mean, of the less common health complaints (incidence less than 1/1000) was calculated. RESULTS: Primary care doctors attended at greater frequency than 1/1000 all diseases of eyes, ears, mastoids (except salpingitis) and menstrual disorders codifiable under the classification CIPSAP; almost all the respiratory, skin and locomotive diseases, and more than half of the circulatory, genito urinary, digestive and endocrine-metabolic diseases. Incidence was less than 1/1000 in all the malignant tumours and contagious diseases, except viral hepatitis and tuberculosis in the urban setting. CONCLUSIONS: Primary care doctors do not often attend certain serious diseases, which are nevertheless present in many differential diagnoses (malignant tumours). This should be borne in mind in the training strategies aimed at maintaining doctor's diagnostic skills. PMID- 10372461 TI - [Scientific papers at scientific conferences: some proposals to improve them]. PMID- 10372462 TI - [Measurement of cardiovascular risk in primary care]. PMID- 10372463 TI - [Controversy in care of climacteric women]. PMID- 10372464 TI - [Academic performance of children with chronic renal failure]. PMID- 10372465 TI - [Prescription use by specialist care in the Toledo Health Area]. PMID- 10372467 TI - Cognitions in generalized anxiety disorder and panic disorder patients. A prospective approach. AB - Self-observations of cognitions during episodes of anxiety were examined in 38 patients with generalized anxiety disorder and 36 patients with panic disorder. Two independent observers who where blind to the diagnoses categorised the cognitions. The inter-rater reliability was high (mean kappa 0.82). The GAD patients had significantly more cognitions in the following categories: interpersonal confrontation, competence, acceptance, concern about others and worry over minor matters, while the PD-patients had significantly more cognitions in the physical catastrophe category. Furthermore, GAD-patients with a comorbidity of social phobia reported more cognitions regarding social embarrassment than did GAD-patients with other or no (axis-I) comorbidity. The results of this study support the cognitive theory regarding the cognitive specificity of anxiety disorders. The implications of these results are discussed, along with the issues of reliability and validity of the instrument used. PMID- 10372466 TI - Depressogenic cognitive styles: predictive validity, information processing and personality characteristics, and developmental origins. AB - Two of the major cognitive theories of depression, the theory of Beck [Beck, A. T. (1967). Depression: clinical, experimental and theoretical aspects. New York: Harper & Row. and Beck, A. T. (1987) Cognitive models of depression. Journal of Cognitive Psychotherapy: an International Quarterly, 1, 5-37] and the hopelessness theory [Abramson, Metalsky, & Alloy, (1989) Hopelessness depression: a theory-based subtype of depression. Psychological Review, 96, 358-372], include the hypothesis that particular negative cognitive styles increase individuals' likelihood of developing episodes of depression, in particular, a cognitively mediated subtype of depression, when they encounter negative life events. The Temple-Wisconsin Cognitive Vulnerability to Depression (CVD) project is a two site, prospective longitudinal study designed to test this cognitive vulnerability hypothesis, as well as the other etiological hypotheses of Beck's and the hopelessness theories of depression. In this article, based on CVD project findings to date, we review evidence that the hypothesized depressogenic cognitive styles do indeed confer vulnerability for clinically significant depressive disorders and suicidality. In addition, we present evidence regarding moderators of these depressogenic cognitive styles, the information processing and personality correlates of these styles and the possible developmental antecedents of these styles. We end with a consideration of future research directions and the clinical implications of cognitive vulnerability to depression. PMID- 10372468 TI - Assumptions in borderline personality disorder: specificity, stability and relationship with etiological factors. AB - The specificity and stability of a set of assumptions hypothesized to be characteristic of Borderline Personality Disorder (BPD) was investigated. BPD patients (n = 16) were compared to cluster-C personality disorder patients (n = 12) and to normal controls (n = 15). All subjects were female and diagnosed with SCID-I and -II. Subjects rated a short version of the Personality Disorder Beliefs Questionnaire (PDBQ), with six sets of 20 assumptions each, hypothesized to be characteristic of avoidant, dependent, obsessive-compulsive, paranoid, histrionic and borderline personality disorder. The BPD assumptions (Cronbach alpha = 0.95) proved to be the most specific to BPD patients. Subjects rated the shortened PDBQ again after viewing an emotional video fragment one week later. Despite increased negative emotions, the PDBQ ratings remained relatively stable. Confirming the cognitive hypothesis, regression analyses indicated that the BPD assumptions mediate the relationship between self-reported etiological factors from childhood (sexual abuse and emotional/physical abuse) and BPD pathology assessed with the SCID-II. It is suggested that a set of assumptions is characteristic of BPD, and is relatively stable despite the instability of the behaviour of people diagnosed as having BPD. PMID- 10372469 TI - An experimental investigation of the role of safety-seeking behaviours in the maintenance of panic disorder with agoraphobia. AB - This study evaluates the hypothesis that safety-seeking behaviours play an important role in maintaining anxiety because they prevent patients from benefiting from disconfirmatory experience. Patients suffering from panic disorder with agoraphobia carried out a behaviour test, closely followed by an experimental session, which included a brief (15 min) period of exposure during which participants either stopped or maintained within-situation safety-seeking behaviours. When the behaviour test was repeated within two days, patients who had stopped their safety-seeking behaviours during the experimental session showed a significantly greater decrease in catastrophic beliefs and anxiety than those who had maintained safety-seeking behaviour. This difference was also reflected in questionnaires measuring clinical anxiety. These results are consistent with the cognitive hypothesis. PMID- 10372470 TI - Psychopathological correlates of self-reported behavioural inhibition in normal children. AB - The present study examined the relationship between self-reported behavioural inhibition and psychopathological symptoms in a sample of 152 children aged 12-14 years. Children were provided with a definition of behavioural inhibition and then asked to classify themselves as low, middle or high on behavioural inhibition. Furthermore, children completed questionnaires of worry, depression and anxiety symptoms. Results showed that children who endorsed the high behavioural inhibition category had elevated levels of anxiety, worry and depression compared to children who endorsed the low or middle behavioural inhibition categories. Moreover, children high on behavioural inhibition more frequently reported anxiety disorders symptoms in the subclinical range. These findings fit well with those of previous studies on behavioural inhibition. PMID- 10372471 TI - Preliminary tests of a cognitive model of generalized anxiety disorder. AB - Although worry is the central feature of Generalised Anxiety Disorder (GAD), little is known about the factors that contribute to pathological or problematic worry. In a recent cognitive model of GAD, Wells, A. (1995) proposed that negative appraisal of worrying itself (meta-worry or type 2 worry) should be distinguished from other types of worrying (type 1 worry). A central feature of this model is the idea that individuals with GAD hold rigid positive beliefs about the usefulness of worrying as a coping strategy. However, these individuals also hold negative beliefs and appraise worrying as uncontrollable and dangerous. This combination of cognitions and associated responses leads to an increased frequency and generality of worrying, and thus to the pathological worry characteristic of GAD. This paper reports a preliminary test of the hypothesis that meta-worry contributes to problematic and pathological worrying, and this relationship is independent of the frequency of other types of worry. In testing for associations between worry dimensions we controlled for overlaps with Trait anxiety, and the controllability of worrying. Results of a series of regression analyses support the hypothesis that pathological worry is associated with meta worry and this association is independent of Trait-anxiety and type 1 worry. The clinical implications of these data are briefly discussed. PMID- 10372472 TI - Some methodological issues in assessing attentional biases for threatening faces in anxiety: a replication study using a modified version of the probe detection task. AB - Various versions of the probe detection task have been developed to assess attentional biases in anxiety and there is debate about their relative merits in terms of reliability and sensitivity to such biases. The present study used a pictorial version of the probe detection task to examine attentional biases for emotional facial expressions. The main aims were (1) to see if our previous finding of greater vigilance for threatening faces in high than low trait anxiety could be replicated [Bradley, B. P., Mogg, K., Falla, S. J., & Hamilton, L. R. (1998). Attentional bias for threatening facial expressions in anxiety: manipulation of stimulus duration. Cognition and Emotion, in press] and (2) to examine whether the same pattern of results and a similar effect size, would be obtained using a 'probe position' task (i.e. where is the probe?), rather than the 'probe classification' task (i.e. what is the type of the probe?) used by Bradley et al. (1998). The probe position task produced similar results to those obtained from the probe classification task, so providing further evidence of an anxiety-related attentional bias for threatening faces. Results also indicated that, for non-clinical participants, the probe position task yielded faster overall RTs, fewer errors and a similar effect size, compared with the probe classification task. Implications for the assessment of attentional biases in non clinical and clinical samples are discussed. PMID- 10372473 TI - 2nd Balkan Congress of Medicine and Dentistry for Students and Young Doctors. Plovdiv, Bulgaria, October 1998. Abstracts. PMID- 10372474 TI - [Knee endoprostheses--advances and questions]. AB - Given the complex biomechanical situation of the knee joint, and the peculiarities of the individual patient, implantation of a knee joint endoprosthesis makes great demands on both the implant and the surgeon. The correct choice of implant from among unconstrained, semiconstrained and constrained types of endoprosthesis, as well as from among cemented, hybrid and uncemented anchorage, is essential. In addition, a meticulous preoperative analysis of knee and leg axes, and appropriate detailed presurgical planning is a must, as is uncompromisingly intensive postoperative physiotherapy. Clinical experience identifies the patella, instability and axis deviations as the major problems of knee arthroplasty. PMID- 10372475 TI - [Knee-preserving oprations in gonarthrosis]. PMID- 10372476 TI - [Chronic constipation in the aged. Consensus conference of the German Society for Geriatrics]. PMID- 10372477 TI - [Chronic pain: prevention by psychologic management]. PMID- 10372478 TI - [Diagnosis of damage: how far does the obligation of medical care go?]. PMID- 10372479 TI - [Spironolactone advantageous to heart failure patients]. PMID- 10372480 TI - The William Guy Memorial Lecture. Dental education: implementing the continuum. PMID- 10372481 TI - Biological side effects to materials used in dentistry. AB - This review paper outlines the potential adverse reactions that may occur through the use of dental materials. Reference is made to documented case reports of side effects. Toxic reactions to permanent dental materials are unlikely, because these materials are developed to be inert and stable and the amount of leachable components is very small. Allergic reactions, on the other hand, are feasible because they may be initiated by minute amounts of the allergen in a sensitised individual. The media in some countries have presented individual patients who allegedly have general symptoms that were relieved by the removal of restorations. A characterisation of these self-referred patients shows that many of them have mental disorders. PMID- 10372482 TI - Small bowel volvulus: a review. AB - Small bowel volvulus is a rare but life-threatening surgical emergency. The aetiology may be primary, as is often seen in Africa and Asia, while in Western countries other predisposing conditions usually initiate the volvulus. Early preoperative investigation and expedient surgical treatment is required if bowel infarction is to be prevented. Central abdominal pain resistant to narcotic analgesia should heighten the suspicion of the diagnosis. The diagnostic value of computerised tomography (CT) scanning in such situations has been emphasised. If the bowel is infarcted resection is required, but the optimum treatment for cases with viable small bowel is uncertain, the alternatives either being resection, fixation, or simple derotation. PMID- 10372483 TI - Partial nephrectomy for renal tumours: the Singapore General Hospital experience. AB - From January 1991 to August 1998, 220 radical nephrectomies were performed for renal cell carcinoma (RCC). During the same period, 27 patients underwent partial nephrectomy for their renal tumours. These included 19 male and 8 female (mean age, 54; range, 35-75). Their clinical presentation, diagnostic modalities and surgical outcome were evaluated. The lesions included 18 RCCs, 7 angiomyolipomas (AMLs), 1 oncocytoma and 1 dysoncogenetic renal tumour. Only 8 patients had specific urological symptoms. Computerised tomography (CT) scan was diagnostic in 78% of cases. Tumour size ranged from 15-50 mm for RCC and 30-190 mm for AML, respectively. Operative time averaged 92 minutes (range: 35-145). The hospital stay ranged from 3 to 25 days (mean 11). Complications occurred in four cases (14.8%); there was one death (3.7%). No tumour recurrence was detected during a mean follow up of 20 months. None of the patients developed significant renal impairment. Partial nephrectomy is feasible in small RCC and some large AML, and can be offered in selected patients. PMID- 10372484 TI - Surgical patients with methicillin resistant staphylococcus aureus infection: an analysis of outcome using P-POSSUM. AB - The significance of MRSA infection in surgical patients was studied using the P POSSUM scoring system. All surgical patients undergoing operation between 1/10/96 and 30/09/97 were prospectively scored using P-POSSUM. A subset of these patients with MRSA infection was analysed using P-POSSUM predicted mortality. Physiological and operative severity scores were compared with non-MRSA surgical patients and length of hospital stay with P-POSSUM matched non-MRSA controls. Thirty of the 1,132 patients were MRSA positive and of these five died, giving a P-POSSUM observed/expected deaths ratio of 1.7 (not significant; 95% CI -0.24 to 0.10). The P-POSSUM physiology score of 30 MRSA positive patients, compared with the non-MRSA group (n = 1102), was significantly more severe (20.9 v/s 17.4; 95% CI 1.09 to 5.95) as was the operative severity score (15.6 v/s 9.2; 95% CI 4.40 to 8.42). The length of stay for surviving MRSA positive patients was significantly longer than P-POSSUM matched controls. MRSA infection in surgical patients does not increase mortality. However, patients who contract MRSA infection are more debilitated and have undergone a greater surgical insult. PMID- 10372485 TI - One-stage procedure in the management of acute sigmoid volvulus. AB - In a descriptive prospective study, twenty-seven patients with sigmoid volvulus and three with ileosigmoid knotting had primary resection of the redundant sigmoid colon with immediate anastomosis after intraoperative antegrade colonic irrigation. There was no clinical anastomotic leak nor mortality in any of our patients. Superficial wound infection occurred in four patients (13.3%). Intraoperative colonic irrigation time ranged between 25 to 50 minutes with a volume of saline/Hartmann's required to achieve a clean colon ranging between 1.5 to 5.0 litres. The duration of hospital stay ranged between 7 and 14 days. The result of this study suggests that resection of acute sigmoid volvulus and primary anastomosis after antegrade intraoperative colonic lavage is safe provided the patient is reasonably fit. PMID- 10372486 TI - Local anaesthesia: to warm or alter the pH? A survey of current practice. AB - Warming or buffering local anaesthetic solutions prior to injection has been shown to reduce the pain experienced by patients. Although clinical practice varies widely, this practice has been commonplace within dental surgery for many years. A postal survey was carried out to ascertain the current practices of a group of general surgeons and maxillofacial surgeons with regard to their local anaesthetic technique. Significant differences existed between the two groups in both their practice and knowledge of this subject. PMID- 10372487 TI - Calman, venous surgery and the vascular trainee. AB - BACKGROUND: Surgical training in Great Britain is undergoing inevitable changes to accommodate the processes of Higher Surgical Training. Junior surgeons have long argued that their training experiences have been haphazard or without satisfactory supervision. With the advent of changes following the Calman Report and the implementation of the Vascular Surgical Society recommendations, we have audited the venous surgical experience of vascular trainees in Great Britain. METHODS AND RESULTS: Questionnaires were sent to 90 vascular surgical trainees achieving an overall 76.7% response rate (n = 69). Just under half of the trainees had spent more than 12 months on a pure vascular firm. The majority of trainees had received formal training in sapheno-femoral junction ligation and sapheno-popliteal junction ligation. However, several areas of training were deemed insufficient at both the junior and senior trainee levels. Very few trainees gain instruction on deep venous surgery and the techniques of microsclerotherapy. CONCLUSIONS: Despite the participation of trainees in specialised vascular units, current training schedules fail to cover the field of venous surgery adequately. Training by vascular specialists needs greater focus and should be tailored to the trainee's experience on entry to their firm. PMID- 10372488 TI - Can the orthopaedic surgeon forget the proctoscope? The proctoscope as tissue protector during intra medullary nailing. AB - We report the value and usefulness of the proctoscope in one of the modern orthopaedic procedures. We have found the proctoscope more effective than the currently available instruments as tissue protector during intramedullary nailing. We briefly describe our technique of using the proctoscope and discuss its advantages. PMID- 10372489 TI - Use of fistula director to enlarge the port site opening to retrieve a stone packed bulky gall bladder during laparoscopic cholecystectomy: a simple and safe technique. AB - Retrieval of the gall bladder through the port site opening is technically difficult and challenging when it is bulky either due to packed multiple calculi or a large solitary calculus. We describe in this article a simple technique that involves enlargement of the port site opening to facilitate the gall bladder removal in these difficult situations, using a scalpel and a fistula grooved director. PMID- 10372490 TI - Do patients with acute abdominal pain wait unduly long for analgesia? AB - A prospective audit of 100 emergency admissions was carried out to determine local surgical practice for analgesia administration in patients with acute abdominal pain. The main outcome measure investigated was waiting time for analgesia and how this was influenced by (i) severity of pain, (ii) clinical diagnosis, (iii) clinical setting. The data were correlated with the results of a questionnaire on timing of analgesia. Forty percent of patients received analgesia within 1 h, 17% between 1-2 h, and 43% 2-22 h after admission. Mean waiting time was 2.3 h with severe pain (n = 84) vs. 6.3 h with moderate pain (n = 16, p < 0.0001, Mann-Whitney). Clinical diagnosis did not influence timing of analgesia. Fifty-seven per cent received analgesia in the Accident and Emergency (A&E) department with a mean wait of 60 min, whereas 43% admitted to the ward without analgesia in the A&E department waited an average of 5.7 h for pain medication (p < 0.0001; Mann-Whitney U-test). This was at variance with local surgical opinion that favoured early analgesia administration (yes-88%), in the absence of a firm diagnosis (yes-79%), although 38% stated that analgesia might mask physical signs. In conclusion, a substantial cohort of patients with acute abdominal pain (43%) wait too long for analgesia. Delays are due to omission of analgesia in A&E, and reluctance of junior staff to administer analgesia for fear of masking physical signs. Clinical guidelines for pain medication in acute surgical emergencies are warranted. PMID- 10372491 TI - Completion of the journey of care: Scottish Audit of Surgical Mortality (SASM). AB - The ideal must be to audit all deaths as part of routine surgical and anaesthetic practice. We have examined the level of compliance of surgeons and anaesthetists participating in the Scottish Audit of Surgical Mortality (SASM) for the audit's first three years. The audit has achieved more than 92% compliance, unchanged over a three year period. As a consequence it can be regarded as a truly routine part of surgical and anaesthetic practice within Scotland. PMID- 10372492 TI - Laparoscopic cholecystectomy. AB - Laparoscopic cholecystecomy is now the gold standard for the treatment of gallstones. When it was first introduced there were some concerns about its safety owing to its rapid adoption by untrained surgeons. However, when a careful, correct technique is employed, the operation is extremely safe. This article describes a well established technique for laparoscopic cholecystectomy, which has been evolved over several years in a centre that has concentrated on the development of laparoscopic surgery. PMID- 10372493 TI - Effect of activity levels on polyethylene wear in Charnley low-friction arthroplasty. AB - Polyethylene wear debri has been implicated as a major cause of aseptic loosening following total hip replacement. The purpose of this study is to classify patient activity levels following total hip replacement and determine if it has an effect on radiographic wear of the socket. 97 patients (115 hips) with osteoarthritis who underwent Charnley low-friction arthroplasty with a minimum of 10 year follow up were selected. Nine patients had died. Eighty-eight questionnaires were sent of which 59 replied. However, only 40 patients (45 hips) with a complete set of x rays were finally studied. Activity levels were classified according to post operative occupation and activities into 'active' and 'less active' groups. Radiographic wear was measured by studying the post-operative and the 10-year review radiograph. There was a significant difference in acetabular wear between the active and the less active group (p = 0.024). No correlation was found between weight and acetabular wear (p = 0.475) or between age and acetabular wear (p = 0.278). It was concluded, therefore, that surgeons who advise their patients to avoid heavy or high intensity activities for prolonged periods, after Charnley low-friction arthroplasty, are justified in this approach. PMID- 10372494 TI - Isolated iliac artery aneurysms with associated hydronephrosis. AB - An isolated iliac artery aneurysm is where there is aneurysmal dilatation of one or more branches of the iliac system, with no associated dilatation of the aorta. Such aneurysms are rare and comprise 1% of all intra-abdominal aneurysms. The signs and symptoms of such an aneurysm are influenced by its concealed location within the bony pelvis. Awareness of these special characteristics improves the chances of early diagnosis and proper treatment before possible rupture. We present the clinical and radiological features of three such aneurysms. Ultrasound was the first imaging modality to be performed. Ipsilateral hydronephrosis was demonstrated in each case, this lead to imaging the pelvis and the correct diagnosis. We review the clinical and radiological literature and conclude that the pelvis should be imaged in all cases of unexplained hydronephrosis. PMID- 10372495 TI - Migration of three endoclips following laparoscopic cholecystectomy. AB - We present a well-documented case report on migration of the haemostatic clip, along with the two clips that had originally been applied on the cystic duct, into the bile duct forming the nidus of a stone after laparoscopic cholecystectomy--a unique occurrence heretofore unreported. The English language literature on the subject is reviewed. The most likely predisposing factor is improper or erratic application of clips resulting in bile leakage, either subclinical or overt. Secure and correct placement of clips could help to prevent this complication. PMID- 10372496 TI - A lethal ectopic denture: an unusual case of sigmoid perforation due to unnoticed swallowed dental plate. AB - We describe a case of generalised peritonitis due to sigmoid perforation caused by an unnoticed swallowed dental plate during sleep three months previously. PMID- 10372497 TI - Management of femoral and tibial disphyseal fractures. PMID- 10372498 TI - The use of rectosigmoid stents in the management of acute large bowel obstruction. PMID- 10372499 TI - Local recurrence following rectal resection for cancer. PMID- 10372500 TI - Foodborne botulism associated with home-canned bamboo shoots--Thailand, 1998. AB - On April 13, 1998, the Field Epidemiology Training Program in the Thailand Ministry of Public Health (TMPH) was informed of six persons with sudden onset of cranial nerve palsies suggestive of botulism who were admitted to a provincial hospital in northern Thailand. To determine the cause of the cluster, TMPH initiated an investigation on April 14. This report summarizes the results of the investigation, which indicate that the outbreak was caused by foodborne botulism from home-canned bamboo shoots. PMID- 10372501 TI - Trends in HIV-related sexual risk behaviors among high school students--selected U.S. cities, 1991-1997. AB - Despite recent decreases in sexual risk behaviors among high school students nationwide, human immunodeficiency virus (HIV) infection was the seventh leading cause of death among persons aged 15-24 years in the United States during 1997. To determine whether the prevalence of HIV-related sexual risk behaviors among high school students also has decreased in certain urban areas heavily affected by the epidemic, CDC analyzed data from Youth Risk Behavior Surveys (YRBS) conducted in 1991, 1993, 1995, and 1997 in eight large-city school districts: Boston, Massachusetts; Chicago, Illinois; Dallas, Texas; Fort Lauderdale, Florida; Jersey City, New Jersey; Miami, Florida; Philadelphia, Pennsylvania; and San Diego, California. This report summarizes the results of this analysis, which indicate that, from 1991 to 1997, the percentage of high school students engaging in HIV-related sexual risk behaviors decreased in some U.S. cities. PMID- 10372502 TI - Illnesses associated with occupational use of flea-control products--California, Texas, and Washington, 1989-1997. AB - Dips, shampoos, and other insecticide-containing flea-control products can produce systemic illnesses or localized symptoms in the persons applying them. Although these products may pose a risk to consumers, they are particularly hazardous to pet groomers and handlers who use them regularly. Illnesses associated with flea-control products were reported to the California Department of Pesticide Regulation, the Texas Department of Health, and the Washington State Department of Health, each of which maintains a surveillance system for identifying, investigating, and preventing pesticide-related illnesses and injuries. This report describes cases of occupational illnesses associated with flea-control products, summarizes surveillance data, and provides recommendations for handling these products safely. PMID- 10372506 TI - Cutaneous sensory spots and the "law of specific nerve energies": history and development of ideas. AB - By use of suitable methods, different spots on the skin surface can be shown to be selectively sensitive to one of four sensory qualities in decreasing order of density: pain, touch, cool and warm. The presence of such spots was observed virtually simultaneously in the early 1880s by three independent investigators. Two papers on punctuate sensitivity of the skin were published in 1882 and 1883 by Magnus Blix of Uppsala University in Sweden; three papers were published in 1884 by Alfred Goldscheider, a German army doctor; and one was published in 1885 by Henry Donaldson of Johns Hopkins University in Maryland. Donaldson's findings originated from a serendipitous observation. In contrast, Blix's and Goldscheider's experiments were based on Johannes Muller's concept of "specific sense energies" and the extension of this idea to sensory qualities (the law of "specific nerve energies") by others, including Hermann von Helmholtz. The discovery of different types of sensory spots had considerable influence on other researchers of the period, including Max von Frey, but has only recently been substantiated by electrophysiological experiments. PMID- 10372507 TI - GABA-ergic neurons and the neurobiology of schizophrenia and other psychoses. AB - There are a number of disorders in the Diagnostic and Statistical Manual of Mental Disorders (DSM IV) that are characterised by having psychotic symptoms as the defining feature [17]. The narrowest definition of psychosis is restricted to delusions or prominent hallucinations, with the hallucinations occurring in the absence of insight into their pathological nature. Schizophrenia is the most prevalent form of psychosis, but this may also occur due to other medical conditions (e.g., Prader-Willi syndrome, epilepsy), in the early post-partum period, at menopause, and as a result of drug use. This article attempts to draw together an underlying causation across the various forms of psychotic disorder and, by integrating this with what is known about the genetics, neuroanatomy and neuropharmacology of the positive symptoms in schizophrenia, produce a broader understanding. At the cellular level, gamma-aminobutyric acid (GABA)-ergic interneurons are a common feature in psychotic states, and are a principal focus for serotonin and dopamine innervations, as well as playing an important role in cortical development. At the systems level, prefrontal and medial temporal cortices are implicated with activity levels out of synchrony in schizophrenics. How these vast areas of disparately functioning cortical networks are "bound" together to provide coherent conscious experiences is again a function of GABA ergic interneurons. These interneurons have highly divergent inhibitory projections to large numbers of pyramidal neurons and are themselves synchronised by the ascending dopamine and serotonin innervations. PMID- 10372508 TI - A working memory "theory of relativity": elasticity in temporal, spatial, and modality dimensions conserves item capacity in radial maze, verbal tasks, and other cognition. AB - It is remarkable that working memory (WM) capacity for numbers of items remains modest, at approximately 7+/-2 (the so-called "magical number"), across a wide variety of kinds of material. Indeed, consideration of radial maze studies together with more traditional memory research shows that WM capacity remains fairly constant whether the items are verbal or visuospatial, and that this same capacity is true of other species as of humans. In contrast to their limited numerousness, WM items are extremely flexible in ways that are here brought under the heading of "dimensionality." Therefore, the physical items represented in WM, can vary widely in any quantitative characteristic and in the temporal pace at which they are encountered. Combinatorial considerations suggest that WM numerousness results from evolution of a middle ground between a sterile parsimony and an overwhelming excess, for organizing neurocognitive associations. Such natural selection seems likely to have worked opportunistically to yield diverse characteristics of neuronal tissue, from subcellular components to properties of ensembles, which converge on the required cognitive properties of WM. Priming and implicit memory may play supporting roles with WM. These intermediate-term memory phenomena allow certain kinds of background information to be accumulated at higher volume than seems possible from the textbook, "modal model" of memory. By expediting attentional focus on subsets of information already in long-term memory, priming may help WM chunks to emerge in limited number as appropriately scaled "figures" from the primed "ground." The larger neuronal dynamic patterns that embody these cognitive phenomena must regulate their microscopic component systems, automatically selecting those having parameters of temporal persistence, rhythm, and connectivity patterns that are pertinent to the current task. Relevant neural phenomena may include "Hebbian" associativity and persistence of firing patterns in prefrontal or hippocampal neurons. A conceivable basis for scaling and normalizing WM representations, along arbitrarily long or short ranges of any cognitive dimension, involves harmonic multiplier relationships among brain electrical rhythms and/or among topographical spatial periodic representations. PMID- 10372509 TI - Effects of mu, kappa, and delta opioid receptor agonists and antagonists on rat hypothalamic noradrenergic neurotransmission. AB - We have investigated the effects of specific mu-, kappa-and delta-opioid receptor agonists and antagonists on the hypothalamic noradrenergic neurotransmission and on luteinising hormone (LH) release in the ovariectomised and steroid-primed rat. The opioid agents were infused intracerebroventricularly under ketamine anaesthesia and blood samples collected at hourly intervals on the afternoon of the anticipated LH surge. At the end of the experiment, the rats were decapitated and the medial preoptic area, suprachiasmatic nucleus, median eminence and arcuate nucleus surgically isolated by micropunch. The concentrations of noradrenaline (NA) and its metabolite (3,4-dihydroxyphenylglycol; DHPG) in these samples was determined by high performance liquid chromatography with electrochemical detection. Plasma LH levels were measured by radioimmunassay. The three opioid agonists reduced concentrations of NA and DHPG in all four hypothalamic areas. These inhibitory effects of the opioid agonists were mostly prevented following coadministration with their respective antagonists. However, naloxone had no significant effect on DHPG levels in any of the hypothalamic regions examined. Plasma LH levels were found to be either low or undetectable in all groups. These results suggest that mu-, kappa- and delta-opioid receptors have inhibitory influence on the hypothalamic noradrenergic neurotransmission around the time of the LH surge. It is thought that the ketamine anaesthesia interfered with LH release. PMID- 10372510 TI - Relationship between modulation of the cerebellorubrospinal system in the in vitro turtle brain and changes in motor behavior in rats: effects of novel sigma ligands. AB - Saturation and competition binding studies showed that the turtle brain contains sigma sites labeled by both [3H]di-o-tolylguanidine (DTG) and [3H](+) pentazocine. There was a significant correlation between the IC50 values of sigma ligands for [3H]DTG sites in the turtle vs. rat brain, suggesting that the sites are comparable in the two species. In contrast, [3H](+)-pentazocine, which primarily labels sigma1 sites in the rodent brain, labels a heterogeneity of sites in the turtle brain. In extracellular recordings from the in vitro turtle brainstem, some sigma ligands enhanced the burst responses of red nucleus (RN) neurons (DTG, haloperidol, BD1031, BD1052, BD1069) while other sigma ligands decreased the burst responses (BD1047, BD1063). Control compounds (turtle Ringer vehicle control, opiate antagonist naloxone, atypical neuroleptic sulpiride) had no significant effects on the RN burst responses recorded from the in vitro turtle brain. The ED50s of the ligands for altering the burst responses in RN neurons from the turtle brain were correlated with their IC50s for turtle brain sites labeled with [3H]DTG, but not [3H](+)-pentazocine; this pattern is identical to that previously reported in rats, where there is a correlation between the potencies of sigma ligands for producing dystonic postures after microinjection into the rat RN and their binding to rat brain sites labeled with [3H]DTG, but not [3H](+)-pentazocine. When the novel sigma ligands were microinjected into the rat RN, dystonic postures were produced by ligands that increased the burst duration of RN neurons in the turtle brain. Novel sigma ligands that reduced the burst responses in the in vitro turtle brain have previously been reported to have no effects on their own when microinjected into the rat RN, but to block the dystonic postures produced by other sigma ligands. Taken together, the data suggest that the opposite effects of the novel ligands in the turtle electrophysiological studies represent the actions of agonists vs. antagonists, and that the directionality of the effects has predictive value for the expected motor effects of the drugs. PMID- 10372511 TI - Polysaccharide mannan components of Candida albicans and Saccharomyces cerevisiae cell wall produce fever by intracerebroventricular injection in rats. AB - The cell wall mannan components of Candida albicans and Saccharomyces cerevisiae produced hyperthermic responses when injected intracerebroventricularly at doses of 10 microg in rats. Indomethacin treatment (5 mg/kg subcutaneously) completely abolished these responses. Serum interferon-gamma, tumor necrosis factor-alpha and interleukin-1beta levels showed an upward trend during the initial phase of the hyperthermic response induced by S. cerevisiae mannan. Meanwhile, serum levels of these proinflammatory cytokines did not increase at all at the initial phase of C. albicans mannan-induced hyperthermia. Histopathological examination of the brain tissue samples revealed no specific change throughout the parenchyma of rats given either mannan. These results indicate that the polysaccharide mannan components of yeasts, regardless of the pathogenicity, produce a pyrogenic response by a direct injection into the brain in rats. This response is not accompanied by proinflammatory cytokine induction in the periphery. PMID- 10372512 TI - Region-specific alterations in the concentrations of catecholamines and indoleamines in the brains of young and old F344 rats after L-deprenyl treatment. AB - The effects of L-deprenyl, a monoamine oxidase-B (MAO-B) inhibitor, on the concentrations of norepinephrine (NE), dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), serotonin (5-hydroxytryptamine), and 5 hydroxyindoleacetic acid (5-HIAA) in medial basal hypothalamus (MBH), substantia nigra (SN), striatum (Str), and nucleus accumbens (NAc) of young (3 month) and old (21 month) male F344 rats were examined after a 7-day wash-out period following 1, 15, or 30 days of deprenyl treatment in young rats and a 9-day wash out period after a 10-week deprenyl treatment in old rats. The brain areas were microdissected and the concentrations of neurotransmitters were measured by High Performance liquid chromatography with electrochemical detection (HPLC-EC). Deprenyl administration following the drug wash-out period increased the concentrations of DOPAC in the SN, Str, and in the NAc of young rats but it was decreased in the NAc of old rats. The concentration of HVA was lower in the Str of young deprenyl-treated rats, and in the Str and NAc of old deprenyl-treated rats, but it was higher in the SN of young deprenyl-treated rats. The concentration of 5-HIAA was increased in the MBH, SN, and in the NAc of young deprenyl-treated rats, but it was decreased in the Str and NAc of old deprenyl treated rats. The concentration of NE was increased in the MBH, SN, Str, and in the NAc of young rats treated with deprenyl and in the MBH of old deprenyl treated rats. The concentration of 5-HT was increased in the SN of young deprenyl treated rats. The concentration of DA increased in the Str of both young and old deprenyl-treated rats. We concluded that a drug wash-out period after deprenyl treatment differentially affects the metabolism of catecholamines and indoleamine depending on the region of the brain and that this effect may be due to variation in the kinetics of MAO inhibition. PMID- 10372513 TI - Effects of the alpha antagonists and agonists injected into the lateral hypothalamus on the water and sodium intake induced by angiotensin II injection into the subfornical organ. AB - The subfornical organ (SFO) and the lateral hypothalamus (LH) have been shown to be important for the central action of angiotensin II (ANG II) on water and salt regulation. Several anatomical findings have demonstrated neural connections between the SFO and the LH. The present experiments were conducted to investigate the role of the alpha-adrenergic antagonists and agonists injected into the LH on the water and salt intake elicited by injections of ANG II into the SFO. Prazosin (an alpha1-adrenergic antagonist) injected into the LH increased the salt ingestion, whereas yohimbine (an alpha2-adrenergic antagonist) and propranolol (a beta-adrenergic antagonist) antagonized the salt ingestion induced by administration of ANG II into the SFO. Previous administration of clonidine (an alpha2-adrenergic agonist) or noradrenaline into the LH increased, whereas pretreatment with phenylephrine decreased the sodium intake induced by injection of ANG II into the SFO. Previous treatment with prazosin and propranolol reduced the water intake induced by ANG II. Phenylephrine increased the dipsogenic responses produced by ANG II, whereas previous treatment with clonidine injected into the LH reduced the water intake induced by ANG II administration into the SFO. The LH involvement with SFO on the excitatory and inhibitory mechanisms related to water and sodium intake is suggested. PMID- 10372514 TI - Nicotine phase shifts the 6-sulphatoxymelatonin rhythm and induces c-Fos in the SCN of rats. AB - The neurotransmitter acetylcholine is not found in the major suprachiasmatic nuclei afferents reported to mediate light effects on entrainment and phase shifts in mammals; however it clearly has some role in the control of circadian rhythmicity. This study examined the effect of the cholinergic agonists nicotine and oxotremorine on (1) the rhythmic production of melatonin using the metabolite, 6-sulphatoxymelatonin as a marker, and (2) the expression of c-Fos protein in the suprachiasmatic nuclei (SCN) of the rat. Nicotine administration (1 mg/kg, s.c.) caused phase delays in the timing of the onset of 6 sulphatoxymelatonin excretion (compared to the pre-treatment night), when administered at circadian time (CT)16 (1.7+/-0.3 h delay) and CT18 (1.7+/-0.2 h delay) but not at CT14 (0.8+/-0.3 h delay), whereas oxotremorine and saline administration had no effect on the timing of the melatonin rhythm. Nicotine administration also caused the induction of c-Fos-like immunoreactivity in the SCN in a dose- and time-dependent manner. Further, pre-treatment with the nicotinic antagonist mecamylamine reduced the number of nicotine-induced c-Fos positive cells in the SCN by 65%. These data indicate that cholinergic neurons may alter the timing of the onset of melatonin excretion by a direct or indirect effect on the SCN possibly mediated by the nicotinic receptor. PMID- 10372515 TI - Induction of presenilins in the rat brain after middle cerebral arterial occlusion. AB - In the present study, we have examined the expression of both presenilins in the rat hippocampus, cortex, striatum, and cerebellum after middle cerebral artery occlusion (MCA-O), an animal model of ischemia. The cortex showed the greatest increase in PS mRNA levels (7-10-fold) at 4 and 8 days posttreatment. Presenilin 1 (PS-1) levels in the contralateral cortex were significantly increased 1 day after MCA-O. In comparison, PS mRNA content was only modestly elevated in the hippocampus and striatum at 4 and 8 days after MCA-O (30-100% changes). Other Alzheimer's disease (AD)-related genes, amyloid precursor protein and apolipoprotein E, are induced in brain injury suggesting that these AD-related genes may well be components of a brain-injury response. Thus, a breakdown in this response via cerebrovascular disease and/or genetic mutation may contribute to AD pathology. PMID- 10372516 TI - Midlatency auditory-evoked potentials in the rat: effects of interventions that modulate arousal. AB - The vertex-recorded P13 midlatency auditory-evoked potential in the rat shows the same characteristics as the P1 potential in the human, namely, sleep-state dependence, rapid habituation and blockade by the cholinergic antagonist scopolamine. The P13 potential appears to be generated, at least in part, by projections of the pedunculopontine nucleus, the cholinergic arm of the reticular activating system. On the other hand, the auditory cortex-recorded P7 potential appears to be of primary cortical origin. Simultaneous recordings from the vertex and the auditory cortex showed that (1) the P13 potential was suppressed by administration of the anesthetics ketamine, pentobarbital or halothane in a dose dependent manner, but the P7 potential was not; (2) the P13 potential was suppressed by intragastric injections of ethanol in a dose-dependent manner, but the P7 potential was not; (3) the amplitude of the P13 potential was negatively correlated with blood ethanol levels; (4) both the P13 and P7 potentials were still present following injections of the neuromuscular blocker pancuronium bromide; and (5) both the P13 and P7 potentials were decreased by diffuse brain injury induced by a weight-drop device in a weight-dependent manner. These findings suggest that the P13 potential is more sensitive than the P7 potential to changes in arousal and that the P13 and P7 potentials are not of myogenic but of neural origin. PMID- 10372517 TI - Magnetic resonance imaging of the abdominal aorta and iliac vessels using combined 3-D gadolinium-enhanced MRA and gadolinium-enhanced fat-suppressed spoiled gradient echo sequences. AB - This study evaluates a combined protocol consisting of breath hold immediate post gadolinium 3-D gradient echo MR angiography and blood pool phase gadolinium enhanced breath hold 2-D fat-suppressed spoiled gradient echo (SGE) sequences in the examination of diseases of the abdominal aorta and iliac vessels. Thirty-two patients with suspected disease of the abdominal aorta, major aortic branches, or iliac vessels underwent MR angiographic study from January 1996 to January 1997. Examinations were performed on a 1.5 T MR imager using 2-D axial SGE, coronal 3-D fast imaging in steady state precession (3-D FISP) following bolus administration of 40 mL of gadolinium, and axial and coronal blood pool phase gadolinium enhanced fat-suppressed SGE. Post-processed data, including 3-D reconstructions using maximum intensity projection (MIP), targeted MIP, and multiplanar reconstruction (MPR) were evaluated. MR findings in all patients were correlated as follows: surgery (13 patients), angiography (11 patients), contrast enhanced CT (3 patients), non-contrast enhanced CT (1 patient), color doppler US (2 patients), and previous MR study (2 patients). MR findings correlated closely with findings at surgery or other imaging studies in 31 of 32 patients. One patient had renal artery occlusion that was misinterpreted as mild stenosis. The following vascular diseases were present: aneurysm disease [10 patients: aortic aneurysm (8 patients), inflammatory aneurysm (2 patients)], thoracoabdominal aortic dissection (2 patients), arteriovenous fistula (1 patient), stenoses and/or occlusion of the abdominal aorta, major aortic branches and iliac vessels [12 patients: stenoses and/or occlusion of the abdominal aorta with stenoses of the iliac vessels (9 patients), renal artery stenosis (2 patients), occlusion of the abdominal aorta (1 patient)], and occluded artery to pancreatic transplant artery (1 patient). Five patients had normal studies. The 3-D FISP technique accurately defined the luminal contours of vessels, allowing precise depiction of vessel stenosis (i.e., renal artery stenosis or common iliac artery stenosis) and clear demonstration of relationship of aortic branch vessels (i.e., renal arteries) to underlying aortic pathology (i.e., aortic aneurysm or dissection). Blood pool phase gadolinium-enhanced fat-suppressed SGE images were useful in the evaluation of the external surface of vessel walls, and providing accurate measurement of aneurysm diameter and other associated vascular entities (i.e., inflammatory aneurysm, left-sided IVC). Targeted MIP or MPR reconstruction were important for assessing stenoses of medium sized vessels such as renal arteries and branches of the iliac arteries, and for identifying accessory arteries. The combination of immediate post gadolinium 3-D FISP and blood pool phase gadolinium enhanced fat-suppressed SGE is useful in the evaluation of the abdominal aorta, major aortic branches and iliac vessels. Immediate post gadolinium 3-D FISP images provides diagnostically useful information regarding vessel luminal contour, while blood pool phase gadolinium-enhanced fat-suppressed SGE provides ancillary information on the vessel wall and surrounding tissue. PMID- 10372518 TI - Comparison of multiple sclerosis clinical subgroups using navigated spin echo diffusion-weighted imaging. AB - The apparent diffusion coefficient (ADC) of tissue provides an indication of the size, shape, and orientation of the water spaces in tissue. Thus, pathologic differences between lesions in multiple sclerosis (MS) patients with different clinical courses may be reflected by changes in ADC measurements in lesions and white matter. Twelve healthy subjects and 35 MS patients with a relapsing remitting (n = 10), benign (n = 8), secondary progressive (n = 8) and primary progressive (n = 9) clinical course were studied. T2-weighted and post-gadolinium T1-weighted images were obtained using a 1.5 T Signa Echospeed magnetic resonance imaging (MRI) system. Diffusion-weighted imaging was implemented using a pulsed gradient spin echo (PGSE) sequence with diffusion gradients applied in turn along three orthogonal directions in order to obtain the average apparent diffusion coefficient (ADCav). Navigator echo correction and cardiac gating were used to reduce motion artifact. ADC maps were derived using a two point calculation based on the Stejskal-Tanner formula. Diffusion anisotropy was estimated using the van Gelderen formula to calculate an anisotropy index. MS lesions had a higher ADC and reduced anisotropy compared with normal appearing white matter. Highest ADC values were found in gadolinium enhancing lesions and non-enhancing hypointense lesions on T1-weighted imaging. MS white matter had a slightly higher ADC and lower anisotropy than white matter of healthy subjects. Lesion and white matter ADC values did not differ between patients with different clinical courses of MS. There was no correlation between lesion ADC and disability. Diffusion-weighted imaging with measurement of ADC using the PGSE method provides quantitative information on acute edematous MS lesions and chronic lesions associated with demyelination and axonal loss but does not distinguish between clinical subtypes of MS. PMID- 10372519 TI - Comparison of T1-weighted spin-echo and 3D T1-weighted multi-shot Echo Planar pulse sequences in imaging the brain at it. AB - The purpose of this study was to evaluate the ability of three dimensional T1 weighted multi-shot Echo Planar Imaging (3D T1w EPI) MR pulse sequence to provide comparable to T1w Spin Echo (SE) results in various diseases of the brain, during shorter acquisition times. Thirty-six patients (aged 30-74 years) with various indications were included in the study. All examinations were performed with a 1T MR scanner with a maximum gradient strength of 15 mT/m. The SE sequence lasted 3 min 50s and the 3D T1w EPI 59s. The quantitative analysis included number of enhancing lesions, signal-to-noise ratio of the enhancing lesions and contrast-to noise ratio (CNR) between enhancing lesions and white matter in both sequences before and after i.v. administration of 0.1 mmol/kg gadopentetate dimeglumine. In addition, the percentage increase of enhancement was measured in each lesion of each sequence. The qualitative analysis included a) conspicuity of the lesions and b) presence of artifacts. The T1w SE sequence was significantly better compared to 3D T1w EPI in all quantitative measurements with the exception of CNR of enhancing lesions before contrast administration and the percentage enhancement. The conspicuity of the lesions did not differ between the two sequences. The EPI sequence presented with significantly more artifacts. We conclude that the 3D T1w EPI sequence could not be used instead of the conventional T1w SE, in routine imaging of the brain. Its overall diagnostic capability, could be useful only in uncooperative patients. PMID- 10372520 TI - Differentiation of hepatic cavernous hemangioma from metastases by rare sequence MR imaging. AB - In this study, in order to differentiate cavernous hemangioma and hepatic metastases, rapid acquisition relaxation enhanced (RARE) sequence was used. First, in vivo measurements of T1, T2 relaxation times and proton density were obtained using T1, T2 calculation protocol (TOMIKON S50, 0.5T) and multipoint techniques. These measurements were made from regions of interest placed over the liver, spleen (because of similarity of relaxation time values between hepatic metastases and spleen) and cavernous hemangioma (HCH). Based on these intrinsic parameters, T2 curves signal intensity of three different tissues were constructed. At TE = 500 ms, the signal intensity of the liver and spleen has been near zero whereas in HCH, the signal intensity remained. As RARE sequence is very similar to spin echo (SE), by replacing effective TE(ETE) = 500 ms in the RARE equation, two dimensional contrast-to-noise ratio (CNR) contour plots were constructed demonstrating signal intensity contrast between liver-spleen, liver Hemangioma for two different scan times (3 min, 7.5 s) and pulse timing. Then, optimal RARE factor and inter echo times were obtained in order to have maximum CNR between liver-Hemangioma and minimum CNR between liver-spleen. These optimal parameters were performed on ten normal and five persons with known HCH. Images showed that in both scan times (3 min, 7.5 s); the liver and spleen were suppressed whereas the HCH was enhanced. The image quality in the scan time of 3 min was better than the scan time of 7.5 s. Moreover, in this study, two different sequences were compared: i) Multi-slice single echo (MSSE) for T1 weighted image ii) RARE (ETE = 80 ms) for T2-weighted image. This comparison was done to show maximum CNR between liver-spleen (metastases) and to choose a better sequence for detecting metastases. CNR in the RARE sequence was more than in the MSSE sequence. PMID- 10372521 TI - Appearance of anterior cruciate ligament autografts in their tibial bone tunnels on oblique axial MRI. AB - The objective of this study was to observe the changing appearance of human anterior cruciate ligament (ACL) grafts in their tibial bone tunnels by MRI using oblique axial images. One-hundred and eight knees in 75 patients were studied by MRI at 1-33 months after arthroscopic ACL reconstructions using double-looped, autogenous semitendinosus and/or gracilis tendons. Knees with poor stability were excluded from this study. The examinations were performed at 0.2T with spin echo proton density and T2-weighted oblique axial images. Appearances of grafts were mainly described on spin echo proton density images based upon time after surgery. The grafts appeared as homogeneous, low signal intensity areas in the bone tunnels at 1 month after the surgery. Ring-shaped low signal intensity areas were observed along the wall of the bone tunnels in the 2- to 3-month group. In many grafts from this group, each tendinous bundle appeared as a low signal area separated by a high signal intensity area. In all cases in the 4- to 6-month group, the thickness of the ring-shaped low signal intensity area had increased, whereas the thickness of the high signal intensity area had decreased. In almost all of the cases, the interior of the bone tunnel gradually became a homologous low signal intensity region by 7 to 12 months after the surgery. According to these results, it is suggested that the maturation of the tendon-bone interface was completed from 6 to 12 months after the ACL reconstruction. PMID- 10372522 TI - Scanning time efficient slinky for non-contrast MRA at low field. AB - To eliminate slab boundary artifact (SBA) for non-contrast multi-slab three dimensional time-of-flight magnetic resonance angiogram (3D TOF MRA), we have previously developed a novel technique, termed SLINKY (Sliding Interleaved kY) acquisition in which a thin slab continuously "walks" along the z-axis while data are acquired in an interleaved fashion along the kY-axis. It has been demonstrated in our earlier works that SLINKY can suppress the SBA without any assumption of blood flow behavior, such as velocity or direction. At the same time, SLINKY keeps the same SNR as conventional multiple overlapping thin slab acquisition (MOTSA). Yet, this method is sensitive to any phase error along the ky axis. In our earlier application of SLINKY, we used navigator echoes to measure and correct the phase errors along the kY axis. The cost of using navigator echo collection is an increase in the imaging time. We therefore propose an improved SLINKY technique which does not use navigator echo collection for correcting phase errors, reducing the imaging time while keeping the same suppression of slab boundary artifacts. The present study demonstrates that by using a specifically designed RF pulse, the navigator echo collection can be avoided without incurring any extra ghosting or SNR reduction in the reconstructed images. PMID- 10372523 TI - MR imaging of flow with locally high spatial resolution. AB - Locally focused magnetic resonance imaging (LF MRI) allows imaging with variable spatial resolution within the field of view (FOV). Because LF MRI uses a priori information to provide locally high resolution in regions with rapid spatial variations in intensity (e.g., blood/tissue interface), it allows accurate reproduction of intense sharp edges in the specimen without blurring and truncation artifacts. This study employs LF MRI for 3D imaging of stationary and pulsatile flow. In the implemented version of LF MRI analytically defined basis functions are used to determine image intensity in regions depicted with low or high resolution. It is demonstrated that LF MRI of flow allows a significant (i.e. 3-4 times) reduction in scan time as compared to conventional FT MRI. It is also shown that LF images of pulsatile flow have a decreased appearance of ghosting artifacts as compared to the images reconstructed by using the conventional method. PMID- 10372524 TI - Comparison of diffusion anisotropy measurements in combination with the flair technique. AB - Fluid-attenuated inversion recovery (FLAIR) technique offers an effective tool to diminish partial-volume averaging effects from cerebrospinal (CSF) signal with in vivo magnetic resonance imaging. CSF-suppressed and unsuppressed direction dependent diffusion-weighted (DW) images are obtained with a DW spin-echo EPI sequence in a single acquisition scheme. Comparison of unsuppressed and CSF suppressed apparent diffusion coefficient (ADC) maps yields consistent values for brain tissue in volunteers when no partial-volume effects are expected, but differs considerably at borders of parenchyma to ventricles and sulci. From theory and phantom studies, a corrected anisotropy index is introduced considering differences of statistical fit errors. Anisotropy of white matter is observed in normal brain of volunteers. Anisotropy index maps reveal destruction of fiber tracts in pathologic areas. Results of a preliminary study on 12 patients with intra-axial tumors indicate an improved delineation of tumor boundaries of FLAIR ADC maps against unsuppressed acquisition. PMID- 10372525 TI - In vivo diffusion characteristics of rat spinal cord. AB - Complete apparent diffusion tensor (ADTs) of spinal cord was measured in vivo in nine rats at 2.0 T. Two rotationally invariant parameters, the trace, which is a measure of the mean diffusivity, and the lattice index (LI), which reflects the degree of orientation coherence of tissue, have been estimated from the ADT. The mean white matter (WM) trace value (3.05 +/- 0.26 mm2/sec) was found to be substantially higher than the gray matter (GM) trace (2.36 +/- 0.39 mm2/sec), in contrast with the published results on fixed, excised cord. Statistically significant anisotropic diffusion was observed in both WM and GM, with greater anisotropy in the WM (LI = 0.67 +/- 0.06) than in the GM (LI = 0.51 +/- 0.05). PMID- 10372526 TI - T2 relaxation of the parotid gland of patients affected by pleomorphic adenoma. AB - The T2 behavior of parotid gland tissue was investigated in 11 patients affected by pleomorphic adenoma. A protocol that was previously set up to define acquisition and post-processing procedures, reaching an accuracy of 2.5% in phantoms and an in vivo long term reproducibility of 0.9-8.5%, was used for the evaluations. The measurements were carried out on a whole body, superconducting imager, using a neck coil as a receiver. Some reference gel samples were imaged together with the patient and used to correct T2 results. The sequence protocol was a multispin-echo, 16 echoes. Signals were fitted with mono and biexponential decay models and an automatic choice of the best model was performed using the two chisquared comparison. Two T2 maps (T2 monoexponential or short T2 component, and long T2 component) and chisquared maps were then produced. Pathologic and normal tissues showed a dominant monoexponential decay with a good level of biexponentiality (16%-27% of total fitted pixels) due to partial volume effects from the liquid content. Concerning the biexponentiality, no significant differences were found between the fitted pixel fraction of normal and pathologic tissue, because the T2 long component of the lesion was related both to the edema and saliva content, but probably the increase in the first compensated the decrease in the second. Chisquared maps showed that most of the lesions presented a monoexponential core surrounded by a biexponential border probably due to a solid component similar to normal tissue with partial volume effects from saliva content. Ninety-five percent confidence intervals for normal tissue were 69.40 87.80 ms (monoexponential relaxation), 38.19-44.67 ms and 285.84-691.28 ms (short and long components of biexponential relaxation). For pathologic tissue they resulted 172.17-275.83 ms, 53.86-89.98 ms and 442.10-814.58 ms. The monoexponential component, mostly present in the core of the lesion, was the parameter that better characterized pathologic tissue. A comparison was performed between normal tissue of patients and normal tissue of volunteers, whose statistics was collected in a previous study with the same evaluation protocol. Results showed no significant differences in the biexponential fitted fractions and the comparison of relaxation times. We conclude that, for tissue characterization, a multiexponential analysis should be carried out in order to improve accuracy and to obtain more reliable results. Moreover, other than relaxation calculations, a topographical analysis of relaxation distribution, using for instance the chisquared maps, might in the future give us more useful information on tissue structure. PMID- 10372527 TI - Correlation between MRI and clinico-pathological manifestations in Lewis rats protected from experimental allergic encephalomyelitis by acylated synthetic peptide of myelin basic protein. AB - Experimental allergic encephalomyelitis (EAE) is an autoimmune disease of the central nervous system which constitutes an accepted animal model for multiple sclerosis (MS). The disease can take an acute or chronic form depending on the injection route, animal strain and nature of the disease-inducing antigen administered. The neuroinflammation associated with the acute form can be detected with T2-weighted, T1-weighted and diffusion MRI, and blood-brain barrier changes can be investigated with Gd-DTPA-enhanced T1-weighted imaging, similar to that of MS patients. A synthetic peptide of myelin basic protein (MBP) encephalitogenic for the Lewis rat (MBP 68-86) was acylated by the attachment of a palmitoyl residue (PAL68-86), and was shown to confer almost complete protection against EAE, when administered to rats before and after an encephalitogenic challenge. In this study, treatment of Lewis rats with PAL68-86 prevented the appearance of clinical signs (p < 0.0001) after challenge with the native peptide (p68-86) in complete Freund's adjuvant (CFA), and reduced considerably the MRI and histopathological signs of the disease (p < 0.0001). Measurement of the gadolinium leakage due to neuroinflammation revealed a significant decrease in permeability from 4.09 +/- 2.1 to 2.95 +/- 1.79% pixels > mean + 2 SD (p = 0.011). Therefore, quantitative MRI measurements correlate very well with the reduced cellular infiltration in the CNS and the absence of clinical signs in the EAE-protected animal. PMID- 10372528 TI - Design of biplanar gradient coils for magnetic resonance imaging of the human torso and limbs. AB - A method is described for design of gradient coils of unconventional geometry for MRI that is based on the superpositions of magnetic fields arising from individual current elements calculated by the Biot-Savart Law. Use of an optimization method based on a genetic algorithm enables a wide diversity in the shapes of coil that can be modeled. To exemplify this a two axis, biplanar gradient set is presented; this geometry offers good access for rectangular objects whilst holding the coils closer to the region of interest than is possible for cylindrical configurations. The inner dimensions of the gradient set were 40.0 x 24.4 x 40.0 cm and the gradient efficiencies were 0.3 and 0.4 mT m( 1) A(-1) in the z- and y- directions respectively over a 15 cm diameter region. Correction of signals arising from regions for which gradient linearity was not optimized was successful for the monotonic region within the set; the largest cuboid from which the MR signal could be processed to produce an undistorted image is of dimensions 36.3 x 17.2 x 24.4 cm. PMID- 10372529 TI - In vivo proton magnetic resonance spectroscopy of diseased prostate: spectroscopic features of malignant versus benign pathology. AB - In vivo Proton Magnetic Resonance Spectroscopy appears potentially useful for non invasive discrimination between benign prostatic hyperplasia (BPH) and prostate carcinoma (PC). Aiming to delimit the range within which spectra from one or the other pathology should occur, and establish extreme spectroscopic features of malignant versus benign prostate disease, we performed endorectal proton MR spectroscopy on 20 patients severely affected of either benign prostatic hyperplasia (BPH) (n = 10) or prostate cancer (PC) (n = 10). They were selected on the basis of the large volume and homogeneity of their lesions, which were histologically confirmed after spectroscopy. Consequently, high-quality short-TE proton spectra with well-resolved metabolite signals, and practically free of volume averaging issues were obtained in all cases. Apart from the typical citrate, creatine, and choline signals of prostate spectra, both BPH and PC spectra showed a peak centered at 3.6 ppm which was assigned to myo-inositol. The intensity of this contribution was found significantly increased in PC cases compared to BPH. Possible relationships between neoplastic transformation and the metabolic pathways in which myo-inositol participates are discussed. Average spectroscopic profiles were calculated for both advanced pathologies, and showed obvious differentiated features. In quantitative terms, the ratio of citrate to choline peak areas as well as that of creatine to myo-inositol appeared as the most convenient to discriminate between advanced PC cases (both ratios below 1.0) and advanced BPH cases (both ratios above 1.0). PMID- 10372530 TI - The study of heat stress in tomato fruits by NMR microimaging. AB - The ripening of the tomato fruit was delayed for several days (average 5 days) by a 1-day heat treatment at 42 degrees C. Ethylene production increased during the first 3 h, but, after 6 h inhibition was almost total in tomato fruit incubated at 42 degrees C. However, recovery of ethylene production was rapid if fruits were returned to a temperature of 25 degrees C after heating. In NMR microimaging, three imaging pulse sequences with different repetition and echo times at 42 degrees C were used to obtain the proton density (TR = 6000 ms, TE = 15 ms), the T1 weighted image (TR = 1000 ms, TE = 15 ms) and the T2-weighted image (TR = 6000 ms, TE = 120 ms). After 12 h heating, the water in locular tissues began to show shorter T1 and T2 values. Though the tomatos were returned to 25 degrees C and preserved one more day, the water having a shorter T2 value in locular tissues, did not change. These results show that tomato fruit do not fully recover from heating even after one day, although ethylene production is recovered almost immediately. For this reason, we suggest that some denaturation event inside the tomato, which goes on after the end of heating, is the cause of the delay in tomato ripening. PMID- 10372531 TI - Reproducibility of magnetic resonance imaging measurements of spinal cord atrophy: the role of quality assurance. AB - A sensitive magnetic resonance imaging (MRI) method to measure spinal cord cross sectional area with the potential to monitor disease progression has recently been developed. As changes in cord area due to disease are usually small, assessment of the reliability of the methodology is essential in serial studies of spinal cord atrophy. The aim of this study was to institute and evaluate a protocol of quality assurance to determine long-term reproducibility of serial studies. Serial MRI of the spinal cord was carried out in five healthy volunteer controls over 1 year. Cross-sectional spinal cord areas were measured in a total of 46 scans. The mean coefficient of variation of all subjects over one year was 1.35%. The intra-observer coefficient of variation for same scan analysis was 0.63%. This study has confirmed high reliability of our serial data over one year and the on-going quality assurance protocol enables continuing evaluation of the reproducibility of results in serial studies. Quality assurance is an essential and practical component of all serial MRI studies, without which the clinical implications of change cannot be reliably evaluated. PMID- 10372532 TI - Hadamard encoding with surface coils for high SNR MR spectroscopy. AB - The advantages of Hadamard over phase encoding in magnetic resonance spectroscopy (MRS) applied with surface coils in the direction perpendicular to the coil are demonstrated experimentally. With the recently introduced, time-shifted adiabatic pulses, the application of Hadamard encoding with surface coils results with almost ideal point spread function for pixels up to a distance of a radius from the coil. Comparison to phase encoding with equal region of interest size shows the significant advantage of Hadamard encoding in slice sharpness, overlapping, and spatial contamination. In addition, since there is no aliasing in Hadamard space, the total experimental time for the same region of interest is much shorter. We conclude that the hybrid of Hadamard encoding in the direction perpendicular to the coil and phase encoding in other directions is the method of choice to obtain reliable high signal to noise ratio MRS in vivo. PMID- 10372533 TI - Variations on the slotted-tube resonator: rectangular and elliptical coils. AB - Two designs (one rectangular, one elliptical) are proposed as efficient alternatives to noncylindrical birdcage RF coils. These designs are based on the slotted-tube resonator and their performance relies on the natural current distribution in the conductors due to the eddy current effects at high frequencies. A Finite element method program, solving the full set of Maxwell's equations, has been employed to accurately characterize and optimize the field homogeneity of the proposed noncylindrical coils. The optimum configuration of each design is presented, taking into account the effect of the RF shield. The proposed designs are compared to several configurations presented in the literature. Two coils (one rectangular, one elliptical) have been constructed and tested in a 0.6 T imaging system. A rectangular coil has been built to operate at 300 MHz. MR images substantiate the usefulness of these coils. PMID- 10372534 TI - Effect of 1.5 T steady magnetic field on neuroconduction of a bullfrog sciatic nerve in a partially active state within several hours after extraction. AB - The bullfrog sciatic nerve within six hours after extraction was determined from the change with time in the action potential and the impedance to be a partially active nerve. Steady magnetic fields have been determined to be ineffective in the neuroconduction in the fresh nerve just after extraction. Although ion motions involved in the neuroconduction in the partially active nerve are not normal, it was confirmed that a 1.5 T steady magnetic field is ineffective in the neuroconduction in the partially active nerve. The obtained results suggest that the neuroconduction in damaged nerves is not affected by the 1.5 T steady magnetic field used in MRI. PMID- 10372535 TI - Telomere dynamics in monkeys: increased cell turnover in macaques infected with chimeric simian-human immunodeficiency viruses. AB - To address the question of how cell turnover is affected by retroviral infections, we used the telomeric terminal restriction fragments (TRFs) as markers of cell replicative history and measured their length in macaques infected with chimeric simian-human immunodeficiency viruses (SHIVs). The TRF lengths of mononuclear cells in 104 samples, including longitudinal samples from nine cynomolgus and ten pig-tailed macaques infected with SHIV, and in samples from 26 uninfected macaques, were quantitated by an improved method, based on two dimensional calibration of DNA sizes, pulsed field electrophoresis, and high resolution Southern blot images. The average TRF lengths of peripheral blood mononuclear cells (PBMCs) from uninfected pig-tailed (14.9+/-1.6 kbp) and cynomolgus (14.1+/-1.8 kbp) macaques were about 3 and 5 kbp longer than those of human infants and 30-year-old adults, respectively. The rate of TRF length shortening in infected pig-tailed macaques was significantly (P = 0.035) higher (2.2-fold) than in uninfected monkeys. The TRFs in SHIV-infected cynomolgus monkeys, which, in general, had lower viral loads than pig-tailed macaques, shortened on average more rapidly (1.6-fold) than in uninfected animals, but the difference was not statistically significant. The TRFs of mononuclear cells from the lymph nodes of two rapidly progressing SHIV-infected macaques that developed AIDS and died also shortened in parallel but somewhat more rapidly than in the PBMCs. These results suggest that the rate of PBMC turnover in macaques could be increased several-fold during infections by immunodeficiency viruses, likely due to immune activation by SHIV antigens. PMID- 10372536 TI - Fatal outbreaks of proliferative enteritis caused by Lawsonia intracellularis in young colony-raised rhesus macaques. AB - Ten juvenile rhesus monkeys (Macaca mulatta) died acutely in two separate disease outbreaks. The animals had segmental thickening of the distal ileum with associated proliferative, rugous appearing mucosae. Microscopically, necrosis, exudative inflammation, mucosal ulceration, and crypt hyperplasia were present. Intracellular organisms were seen histochemically and ultrastructurally, and were confirmed to be Lawsonia intracellularis using a specific immunohistochemical method. Proliferative enteropathic conditions caused by L. intracellularis are reported in an ever-increasing range of hosts, suggesting that the infection may exist unrecognized in an even greater array of species, possibly including man. PMID- 10372537 TI - Biochemistry and haematology values for the baboon (Papio hamadryas): the effects of sex, growth, development and age. AB - A retrospective study evaluated the influence of sex and age on plasma biochemistry and haematology parameters in a captive-bred colony of baboons. Over 1,140 ETDA and heparin blood samples were obtained from 160 clinically normal baboons between the ages of 11 months and 11 years. Data for these blood tests were analysed for the effects of sex, age and sex age interactions. Sex, age and sex age interactions were detected for many plasma biochemistry and haematological parameters. The reference range values for platelets, white-blood cells and mean corpuscular volume and plasma chloride, glucose, total protein and iron were higher (P < 0.01) and red blood cell, plasma sodium, potassium, total CO2, creatinine, urea, total bilirubin, albumin, alkaline phosphate, gamma glutamyl transpeptidase and phosphate were lower (P < 0.01) in the female compared to the male population. Sex age interactions (P < 0.05) were seen with haemoglobin, white blood cells, haematocrit, mean corpuscular volume, sodium, creatinine, urea, calcium, phosphate, total bilirubin, total protein alkaline phosphatase, the liver enzymes and triglycerides. Plasma alkaline phosphatase was highest ( > 800 micro/l) in young juveniles of both sexes; creatinine was higher in older ( > 4 years) compared to younger baboons of the same sex (P < 0.05). Plasma cholesterol and triglycerides were greater (P < 0.01) in young baboons compared to older animals. PMID- 10372538 TI - Presence of platelet-activating factor in rhesus (Macaca mulatta) spermatozoa. AB - Platelet-activating factor [1-O-alkyl-2-acetyl-sn-glycero-phosphocholine; PAF] is a unique signaling phospholipid which has been implicated in a number of biological activities (e.g., reproduction). PAF has been detected in the spermatozoa from a number of laboratory and domestic species, including, but not limited to, rabbit, bovine, and the mouse. The concentration of PAF is inversely related to human (Homo sapien) spermatozoal quality. Additionally, PAF levels are significantly higher in Bolivian squirrel monkey (Saimiri sciureus) spermatozoa obtained during the breeding season than spermatozoa obtained during the nonbreeding season. There are no reports on the presence of PAF in rhesus (Macaca mulatta) spermatozoa. Therefore, the primary objective of this study was to detect the presence of PAF in rhesus spermatozoa. A second objective was to determine if PAF spermatozoa levels differ between animals housed individually (single-caged) versus free-ranging (open corrals). Semen were collected from mature rhesus via electro-ejaculation. Spermatozoa were washed free of ejaculatory plug and quick frozen in PBS. Endogenous lipids were extracted from thawed spermatozoa and ejaculatory plugs then assayed for the presence of PAF by [125I]-radioimmunoassay. PAF was not detected in any ejaculatory plugs. PAF levels were significantly higher (P < 0.01) in spermatozoa obtained from free ranging males (mean: 1.16 pmol/10(6) spermatozoa) than males housed individually in single cage units (mean: 0.53 pmol/10(6) spermatozoa). PAF was present in rhesus spermatozoa. Additionally, PAF levels were higher in spermatozoa obtained from corral-housed animals. Additional studies are warranted to elucidate the role of PAF in spermatozoa function. PMID- 10372539 TI - Efficacy and effects of short- and medium-term contraception in the common marmoset (Callithrix jacchus) using melengestrol acetate implants. AB - This study examines the effect of melengestrol acetate (MGA) implants on reproductive function and various biochemical parameters, ovarian activity, and uterine morphology in ten female common marmosets implanted for either 6-8 or 19 21 months. Measures of body weight, concentrations of urinary glucose and blood liver enzymes were taken. Ovarian activity was assessed by analysis of urinary progestin levels and ultrasound examinations of the ovaries. Ultrasonography was also used to evaluate uterine morphology. MGA was highly effective in preventing pregnancies in the study animals. No changes in biochemical parameters were found; however, seven females developed a substantial weight gain during the study. Follicular development was not suppressed, as indicated by the presence of antral follicles, luteinized structures, and elevated urinary progestin levels. The uteri of the MGA-treated subjects were moderately enlarged with a thickened endometrium that showed a marked change in structural appearance indicative of hypertrophy and decidualization. After implant removal these changes quickly disappeared and all females ovulated within 3 weeks and conceived within 4 months post-treatment. MGA appears to be an acceptable contraceptive in the marmoset, although non-steroidal methods should be evaluated as possible potential alternatives. PMID- 10372540 TI - HLA-DM can partially replace the invariant chain for HLA-DR transport and surface expression in transfected endocrine epithelial cells. AB - The function of HLA class II molecules as peptide presenters to CD4+ T cells depends on the expression of associated molecules such as the invariant chain (Ii) and DM responsible for the correct transport of and high-stability peptide binding to the class II dimers. In organs affected by autoimmune diseases, endocrine epithelial cells express class II molecules, which presumably are involved in the presentation of self-peptides to autoreactive T cells. We have transfected the rat insulinoma cell line RINm5F with different combinations of HLA-DR, Ii and HLA-DM cDNAs and have studied how Ii and DM affect the transport and stability of class II molecules expressed by the different transfectants. Immunofluorescence and biochemical analysis showed that cells transfected with DR and DM in the absence of Ii expressed mostly stable molecules in their surface, and showed a lower accumulation of DR molecules in the endoplasmic reticulum (ER) than cells expressing only DR. This suggests that, in the absence of invariant chain, DM molecules can not only exchange peptides other than class II-associated invariant chain peptide (CLIP) but may also be involved in the transport of class II molecules out of the ER towards the endosomal route. In addition, these data confirm that expression of DR alone or DR+Ii do not allow the formation of sodium dodecyl sulphate (SDS)-stable complexes, that cells expressing DR+Ii have most DR molecules occupied by CLIP and that Ii and DM molecules allow regular routing and peptide loading of class II molecules. PMID- 10372541 TI - HLA class II haplotypes in primary sclerosing cholangitis patients from five European populations. AB - The association of primary sclerosing cholangitis (PSC) to HLA class II genes was studied by comparing patients from five different European populations. Deduced HLA-DRB1, DQA1, DQB1 haplotypes of 256 PSC patients from England, Italy, Norway, Spain and Sweden were compared to those observed in 764 ethnically-matched controls. Increased frequencies of the DRB1*03, DQA1*0501, DQB1*02 (RR=3.0, P<0.00001) and the DRB1*13, DQA1*0103, DQB1*0603 haplotypes (RR=2.4, P<0.0001) were observed in all five patient groups. A total of 16% of the PSC patients were homozygous for the DRB1*03, DQA1*0501, DQB1*02 haplotype compared to 1% of the controls (RR=20, P<0.0001). The DRB1*04, DQA1*03, DQB1*0302 haplotype was significantly reduced in frequency(RR=0.4, P<0.00001). Among Norwegian, Swedish and British patients that did not carry neither the DRB1*03, DQA1*0501, DQB1*02 nor the DRB1*13, DQA1*0103, DQB1*0603 haplotype, an increased frequency of the DRB1*15, DQA1*0102, DQB1*0602 haplotype was observed (RR=2.0, P<0.0001). Thus, PSC was found to be positively associated to three different HLA class II haplotypes (i.e. the DRB1*03, DQA1*0501, DQB1*02, the DRB1*15, DQA1*0102, DQB1*0602 and the DRB1*13, DQA1*0103, DQB1*0603 haplotypes) and negatively associated to one HLA class II haplotype (i.e. the DRB1*04, DQB1*0302 haplotype). PMID- 10372542 TI - CD28/CTLA4 gene region on chromosome 2q33 confers genetic susceptibility to celiac disease. A linkage and family-based association study. AB - Celiac disease (CD) is a common small intestinal injury caused by sensitivity to gliadin in genetically-predisposed individuals. The only susceptibility locus established is the HLA-DQ. We tested whether the chromosomal region of the CD28/CTLA4 genes on 2q33 is linked to CD. These genes encode receptors regulating the T-lymphocyte activation. Recently, this gene region was reported to be linked to the susceptibility to many autoimmune diseases, including insulin-dependent diabetes (IDDM12locus). It is thus an obvious candidate locus also for CD, since the intestinal injury is mediated by the immune system. Genetic linkage between seven marker loci in this gene region and CD was studied in 69 Finnish families. In the multipoint linkage analysis, the highest non-pararametric linkage score (NPL) was 1.75 (P=0.04) for D2S116, suggesting weak linkage for this candidate locus. To evaluate this finding, an additional 31 families were typed for all markers. In the combined set of 100 families the NPL score for marker D2S116 was 2.55 (P=0.006) and for other markers 1.90-2.47 (P=0.029-0.007), supporting genuine linkage at this region. Significantly, locus D2S116 also showed a clear allelic association in these 100 families (P=0.0001). The transmission/disequilibrium test (TDT) for locus D2S116 gave preliminary evidence for preferential maternal non-transmission of allele *136 to patients (TDTmax=8.3; P<0.05). No paternal deviation was found suggesting that the effect of the locus might be mediated by a sex-dependent factor protective against CD. Our results indicate that the CD28/CTLA4 gene region can contain a novel susceptibility locus for CD and support the hypothesis that CD has an immune system-mediated component. Like the HLA, the CD28/CTLA4 genes appear to be associated with genetic susceptibility to various autoimmune diseases. PMID- 10372543 TI - Novel HLA-A and HLA-B alleles in South American Indians. AB - The human leukocyte antigen (HLA) complex includes the most polymorphic genes in humans. More than 600 allelic variants have been described in different populations. The HLA-B locus has contributed the largest number of alleles. Although Native American populations display a restricted number of HLA-alleles, many novel HLA class I alleles have been identified in indigenous communities of Central and South America. We have studied 248 unrelated individuals from three tribes of North-East Argentina and one from South-West Brazil, as well as 80 related individuals from the Brazilian tribe. In the course of this work, we found 8 new B-locus alleles and 2 novel A-locus alleles in these populations. Here we report the nucleotide sequences of A*0219, A*0222, B*3519, B*3520, B*3521, B*3912, B*4009 and B*4803 and we show their relationship with similar alleles. The new alleles B*35092 and B*3518 have been described by us in a previous paper. The possible mechanisms that may have produced these alleles over evolutionary time are discussed. PMID- 10372544 TI - Sequencing of HLA class II genes based on the conserved diversity of the non coding regions: sequencing based typing of HLA-DRB genes. AB - In this paper, we present a novel sequencing based typing strategy for the HLA DRB1, 3, 4 and 5 loci. The new approach is based on a group-specific amplification from intron 1 to intron 2 according to the serologically-defined antigens. For this purpose, we have determined the 3' 500 bp-fragment of intron 1 and the 5' 340 bp-fragment of intron 2 of all serological antigens and their most frequent subtypes. We discovered a remarkably conserved diversity characterized by lineage-specific sequence motifs. This lineage-specificity of non-coding motifs in the 1st and 2nd intron offered the possibility to establish a clear serology-related amplification strategy. The method allows the complete analysis of the 2nd exon and the definition of the cis/trans linkage of sequence motifs by intron-mediated polymerase chain reaction (PCR)-based separation of the haplotypes in nearly all serologically heterozygous samples. In particular, the non-coding variabilities between the DR52-associated DRB1 groups made their independent amplification possible. Thus, compared to the standard procedures using exon-based amplification primers, the groups DR3, DR12, some DR13 alleles (1301, 1302) and the DR14 group could be amplified by specific primer mixes. The DR8 could be amplified with an individual primer mix not co-amplifying the DR12. The DR11 and DR13 did not show any individual motif in intron 1 or intron 2. In order to achieve a separate amplification, they had to be amplified by multispecific primer mixes (DR3/11/13/14; DR3/11/13 or DR11/13/14) excluding the other haplotype. Thus, exclusively the alleles in rare DR11,13 heterozygosities without a DRB1*1301 or 1302 could not be amplified separately. Fourteen primer mixes are used to amplify the specificities DR1-14, and 6 primer mixes for the specificities DR51-53. The sequence homology of the 3' end of intron 1 facilitated the application of only three different sequencing primers for all DRB alleles. PMID- 10372545 TI - A DNA-based detection and screening system for identifying HLA class I expression variants by sequence-specific primers. AB - Molecular methods are now commonplace for HLA typing and they have replaced traditional serological methods in many histocompatibility laboratories. A consequence of reliance on molecular methods using primers or probes based on existing sequence information is that unsequenced or partially-sequenced null, or low expressed variants are not discriminated from expressed alleles. Failure to identify null alleles might have deleterious implications for allogeneic transplants. Expression variants may be classified into two categories: unique mutations and repeat mutations. For example, the alleles A*0303N, A*2409N, and B*1526N have apparently unique mutations. In contrast, repeat mutations may occur frequently at points where unusual nucleotide sequences make accurate DNA replication by DNA polymerases difficult. One example is between nucleotide positions 621-627, where HLA class I alleles may exhibit between three and seven consecutive cytosine residues. Incorrect insertion of an extra cytosine in this region is the cause of expression failure in A*2411N and A*0104N alleles. We hypothesise that insertion of an extra cytosine into the cytosine island between nucleotide positions 621-627 is likely to recur not only in other HLA-A alleles but also in HLA-B and even HLA-C alleles. We describe here a polymerase chain reaction using sequence-specific primers (PCR-SSP) system that can not only detect all previously-sequenced HLA class I expression variants but can also screen for mutations between positions 621-627 in HLA-A, B or C alleles which may give rise to potentially null alleles. Overall, in this study HLA class I expression variants were identified in 5 of 931 tested samples (0.53%). PMID- 10372546 TI - Characterization of a novel HLA-A2 variant, A*02012, in the Southern Thai-Muslim population. AB - Southern Thai Muslims (STM)--from Nakon Si Thammarat, whose ancestors come mainly from Malaysia--constitute more than half of all Thai Muslims which, in total, represent approximately 10% of the country's population. The most common A2 subtypes in STM were A*0203 (n=15), A*0201 (n=8) and A*0207 (n=7). In this study, samples with unexpected amplification patterns were sequenced. Three individuals were indicative of a novel A2 allele, now known as A*02012. PMID- 10372547 TI - Correction of HLA-Cw*0501 and identification of HLA-Cw*0711. AB - We describe a new HLA-C allele, Cw*0711, that differs from Cw*0704 by one residue in the intracytoplasmic region, and correct two errors in the published sequence of Cw*0501. PMID- 10372548 TI - Identification of a new variant, HLA-Cw*1507, differing from Cw*1502 only at the KIR-related dimorphism of codons 77 and 80. AB - We describe here a novel allele, HLA-Cw*1507, identified by polymerase chain reaction using sequence-specific primers (PCR-SSP) and sequence-based typing (SBT). Cw*1507 is similar to Cw*1502 with differences at nucleotide positions 302 (A to G) and 312 (A to C) in exon 2. The substitutions observed in Cw*1507, change codon 77 from AAC (asparagine) to AGC (serine) and codon 80 from AAA (lysine) to AAC (asparagine), compared to Cw*1502. Residues 77 and 80 of HLA-C alleles are located in the alpha 1 domain, where they can influence interaction between antigenic peptides and the T-cell receptor. Also, the dimorphism at these residues from asparagine and lysine to serine and asparagine, respectively, are known to modulate interaction with the natural killer (NK) cell killer inhibitory receptor (KIR). The new HLA-Cw*1507, together with Cw*1502, represents the fourth pair of HLA-C alleles differing only at the KIR-related dimorphic codons 77 and 80. PMID- 10372549 TI - A novel HLA-DR12 allele (DRB1*1206) found in a Korean B-cell line. AB - At least 6 HLA-DRB1*12 alleles have been identified to date with nucleotide polymorphism occurring at codons 37, 57-58, 60, 67, 85 and 87. In this report, we describe the identification of another new HLA-DRB1*12 allele: DRB1*1206. This novel allele was found in an Epstein-Barr virus (EBV)-transformed Korean B-cell line "K-KT" having the HLA-phenotype A3, 24; B44, 61; Cw3; Bw4, 6; DR12, 13 during full-length cDNA isolation for cell line characterization and for production of HLA-DR recombinant proteins. The allele was identified initially by cycle sequencing of subcloned HLA-DRB full-length cDNA. PMID- 10372550 TI - Cell cycle genes in chondrocyte proliferation and differentiation. AB - Coordinated proliferation and differentiation of growth plate chondrocytes controls longitudinal growth of endochondral bones. While many extracellular factors regulating these processes have been identified, much less is known about the intracellular mechanisms transducing and integrating these extracellular signals. Recent evidence suggests that cell cycle proteins play an important role in the coordination of chondrocyte proliferation and differentiation. Our current knowledge of the function and regulation of cell cycle proteins in endochondral ossification is summarized. PMID- 10372551 TI - Analysis of coding sequences for tissue inhibitor of metalloproteinases 1 (TIMP1) and 2 (TIMP2) in patients with aneurysms. AB - Aneurysms are characterized by dilation, i.e. expansion and thinning of all the arterial wall layers, which is accompanied by remodeling of the connective tissue. Genes involved in the regulation of tissue remodeling are therefore candidate genes. We analyzed TIMP1 and TIMP2 coding sequences in 12 individuals with abdominal aortic aneurysms (AAA), one individual with AAA and intracranial aneurysms (IA), four individuals with IA and two clinically unaffected individuals. We identified two nucleotide variants in both the TIMP1 and the TIMP2 coding sequences. All differences occurred in the third base positions of codons and were neutral polymorphisms. A significant difference was observed in the frequency of TIMP2 nt 573 polymorphism between 168 alleles from AAA patients and 102 control alleles. PMID- 10372552 TI - Chondroitin sulphate and heparan sulfate proteoglycan are sequentially expressed in the uterine extracellular matrix during early pregnancy in the rat. AB - Chondroitin sulfate proteoglycan (CSPG) and heparan sulfate proteoglycan (HSPG) are extracellular matrix proteins that regulate cell adhesion, growth, migration, differentiation and gene expression in many systems. In this study, stromal CSPG label was intense within 10 microm of the uterine lumen. From that distance to the myometrium, CSPG was de-expressed. From the time of implantation on Day 6, this pattern was reversed. CSPG was de-expressed from the uterine epithelium to a distance of approximately 10 microm from the uterine lumen. From that region to the myometrium, labeling was homogeneously intense. This finding suggests that CSPG may inhibit attachment and implantation. Heparan sulfate core proteoglycan (perlecan) was increasingly expressed in the uterine epithelium from the time of implantation, commencing in the basement membrane on Day 6 and extending to the apical epithelium and lateral plasma membranes by Day 7. Perlecan thus appears to facilitate trophoblast attachment and implantation. We propose that attachment and implantation is regulated, at least in part, by the selective and sequential expression of CSPG and perlecan. PMID- 10372553 TI - Sequential expression of matrix protein genes in developing rat teeth. AB - Tooth organogenesis is dependent on reciprocal and sequential epithelial mesenchymal interactions and is marked by the appearance of phenotypic matrix macromolecules in both dentin and enamel. The organic matrix of enamel is composed of amelogenins, ameloblastin/amelin, enamelins and tuftelin. Dentin is mainly composed of type I collagen, but its specificity arises from the nature of the non-collagenous proteins (NCPs) involved in mineralization, phosphophoryn (DPP), dentin sialoprotein (DSP), osteocalcin, bone sialoprotein and dentin matrix protein-1 (Dmp1). In this paper, we studied the pattern of expression of four mineralizing protein genes (type I collagen, amelogenin, DSPP and osteocalcin) during the development of rat teeth by in situ hybridization on serial sections. For this purpose, we used an easy and rapid procedure to prepare highly-specific labeled single-stranded DNA probes using asymmetric polymerase chain reaction (PCR). Our results show that type I collagen is primarily expressed in polarizing odontoblasts, followed by the osteocalcin gene expression in the same polarized cells. Concomitantly, polarized ameloblasts start to accumulate amelogenin mRNAs and transiently express the DSPP gene. This latter expression switches over to odontoblasts whereas mineralization occurs. At the same time, osteocalcin gene expression decreases in secretory odontoblasts. Osteocalcin may thus act as an inhibitor of mineralization whereas DSP/DPP would be involved in more advanced steps of mineralization. Amelogenin and type I collagen gene expression increases during dentin mineralization. Their expression is spatially and temporally controlled, in relation with the biological role of their cognate proteins in epithelial-mesenchymal interactions and mineralization. PMID- 10372554 TI - Overview of expression of matrix metalloproteinases (MMP-17, MMP-18, and MMP-20) in cultured human cells. AB - We recently described the cell type distribution of several matrix metalloproteinases (MMP-1 through MMP-16). In this report we extend this study by analysis of three recently described MMPs. PCR primers for MMP-17, MMP-18, and MMP-20 were optimized for use in RT-PCR. The results demonstrate one or more cell lines or tissue that express mRNA for each of these newly described MMPs. PMID- 10372555 TI - Primary structure of the helical domain of porcine collagen X. AB - The entire primary structure of the collagen X helical region is presented, including identification of the extensive post-ribosomal modifications by amino acid sequencing and mass spectrometry. As in collagen I, a single residue of 3 hydroxyproline was identified, but for collagen X this was located near the N terminal end of the helix. Lysine residues in collagen X are extensively hydroxylated/glycosylated: at least 11 sites were localized and shown to be fully glycosylated, exclusively as glucosyl-galactosyl derivatives. The lysine-derived crosslinks, dihydroxylysinonorleucine and hydroxylysinonorleucine, were shown to be present in a 3:2 molar ratio primarily within the C-terminal portion of the helix. PMID- 10372556 TI - Complete genomic structure of the mouse cathepsin K gene (Ctsk) and its localization next to the Arnt gene on mouse chromosome 3. AB - In preparation for gene manipulative experiments in mice we have isolated genomic clones covering the entire murine cathepsin K gene (Ctsk). Sequence analysis revealed that the gene spans 10.1 kb and contains eight exons and seven introns. The sizes of the coding exons 2-7 as well as the pattern of intron sizes are conserved between the human and mouse cathepsin K genes. Genomic organization of the mouse cathepsin K gene also closely resembles those of cathepsins S and L. More than 26% of the Ctsk sequence consists of different repetitive elements. Detailed sequence analysis of 1.7 kb of Ctsk promoter revealed several putative binding sites for transcription factors and two stretches of 280-320 bp which were > 70% homologous with the human cathepsin K promoter. Analysis of genomic clones extending further upstream of the Ctsk gene identified the 3' end of the gene coding for arylhydrocarbon-receptor nuclear translocator (Arnt). This places Ctsk approximately 4.5 kb downstream of Arnt on mouse chromosome 3 at locus 47.9. PMID- 10372557 TI - Human perlecan immunopurified from different endothelial cell sources has different adhesive properties for vascular cells. AB - Perlecan, a major heparan sulfate proteoglycan of vascularized tissues, was immunopurified from media conditioned by human endothelial cells of both arterial and venous origin. The heparan sulfate moiety of perlecan from cultured arterial cells differed in amount and/or composition from that produced by a transformed cell line of venous origin. Both forms of perlecan bound basic fibroblast growth factor with Kd approximately 70 nM. In ELISA experiments, perlecan and its protein core bound to various extracellular matrix components in a manner that was strongly influenced by the format of the assay. Human vascular smooth muscle cells and human endothelial cells adhered to perlecan-coated surfaces, and both cell types adhered better to the venous cell-derived than to the arterial cell derived perlecan. Removal of the heparan sulfate chains abolished this difference and increased the ability of both types of perlecan to adhere vascular cells. Denaturation of perlecan and its protein core also rendered each of them more adhesive, indicating the presence of conformation-independent adhesion determinants in the polypeptide sequence. Their location was investigated using recombinant perlecan domains. Overall, our results represent the first demonstration of human perlecan acting as an adhesive molecule for human vascular cells and suggest that it may play a role in vascular wound healing. PMID- 10372558 TI - Tissue specificity of a new splice form of the human lysyl hydroxylase 2 gene. AB - In this study we present the first report of alternative RNA splicing in a gene for lysyl hydroxylase (LH) in a normal population. This splicing event, which we have observed in the LH2 gene, appears to be tissue specific. The LH2 isoform was recently cloned and sequenced from a human kidney cDNA library and predicted to encode a 737 amino acid protein. In the present study, we have isolated a cDNA for LH2 from human skin fibroblasts that codes for a protein of 758 amino acids, of which 21 amino acids are encoded by a new exon. This 63-bp exon, designated exon 13A, is located between exons 13 and 14 of the originally-described LH2 gene. Amplification of cDNAs by PCR, using primers from exons 13 and 14, showed the presence of two distinct LH2 mRNA populations. A 209-bp transcript was expressed in mRNAs isolated from all tissues examined and was the only transcript expressed in skin, lung, aorta and dura, whereas in mRNAs from spleen, cartilage, liver, kidney, frontal lobe and placenta, an additional shorter 146-bp transcript was amplified. DNA sequence analysis showed that these two mRNAs resulted from the alternative splicing of exon 13A. The transcript containing exon 13A is expressed as the major LH2 form in all tissues except kidney and spleen. Analysis of genomic DNA from skin, placenta and spleen showed that both transcripts were generated from the same LH2 gene. Both upstream (intron 13) and downstream (intron 13A) sequences bordering exon 13A had normal consensus sequences for the acceptor (ag) and donor (gt) splice sites. Preliminary studies indicated that only single transcripts which included exon 13A were amplified from normal fetal skin at different stages of gestation. This suggests that although exon 13A is variably expressed in different tissues, this alternative splicing event is not developmentally regulated. PMID- 10372559 TI - Collagen II containing a Cys substitution for Arg-alpha1-519. Analysis by atomic force microscopy demonstrates that mutated monomers alter the topography of the surface of collagen II fibrils. AB - A recombinant human procollagen II was prepared that contained a substitution of Cys for Arg at alpha1-519 and that was found in five families with early onset generalized osteoarthritis with or without features of a mild chondrodysplasia. Previously, the presence of mutated monomers in mixtures with wildtype collagen II was shown to increase the lag period for fibril assembly. Also, the fibrils were more loosely packed and some thick fibrils lacked a D-periodic banding pattern. Here we re-examined the fibrils using a combination of transmission electron microscopy and atomic force microscopy. The presence of the mutated monomers increased the diameter of the thin filaments that were consistently formed in association with the thick fibrils of collagen II. In addition, the presence of the mutated monomers increased the depth of the gap regions in all fibrils with a distinct D-periodic banding pattern. The results, therefore, may indicate that the mutated monomers formed two or three additional outer layers of monomers in 0D-period staggers on the surface of the fibrils. Apparently, the mutated monomers were bound on the surface through intermolecular disulfide bonds. PMID- 10372560 TI - Laminin-5 promotes adhesion and migration of epithelial cells: identification of a migration-related element in the gamma2 chain gene (LAMC2) with activity in transgenic mice. AB - The effects of laminin-5 and its subunit gamma2 chain on cell adhesion and migration were studied, and a migration-related cis-acting element was identified in the gamma2 chain gene (LAMC2) using promoter-reporter gene constructs in transgenic mice. Intact laminin-5 molecules, but not recombinant gamma2 chain promoted cell adhesion of human keratinocytes and mouse squamous carcinoma cells, indicating that the gamma2 chain does not contain a cellular binding site. However, the gamma2 chain as such is probably involved in the process of cell locomotion, as antibodies against the short arm of the chain inhibited migration of carcinoma cells in an in vitro assay. Further evidence for the involvement of the gamma2 chain in cell migration was obtained by the identification of a cis acting element in a promoter-lacZ reporter gene construct that was active in migratory epithelial cells of healing wounds in mice made transgenic by microinjection of the construct into fertilized oozytes. The migration active element was located in the sequence between -613 and +55. The same construct, and another one containing 5900 base pairs of the 5' flanking region, yielded very limited expression in cells of normal tissues. The limited expression was, however, only observed in epithelial cells of different tissues, i.e. cell types that normally express laminin-5 in vivo. The results show that the sequence between -613 and +55 contains elements that can drive expression during epithelial cell migration and that also partially confers more general epithelium expression. However, elements outside -5900 and +55 are needed for normal epithelium expression of the LAMC2 gene. PMID- 10372561 TI - Changes in a measure of cardiac vagal activity before and after epileptic seizures. AB - In previous studies of the relationship between the cardiac autonomic activity and seizures, assessment of autonomic changes has relied on alterations in heart rate or R-R intervals. We have used a recently developed continuous index of cardiac parasympathetic activity (CIPA) to examine 20 seizures in 10 patients during pre-surgical evaluation in a video-telemetry unit. The patients had localization related seizures due to non-progressive pathology and both complex partial seizures (CPS) and complex partial with secondary generalised tonic clonic seizures (STCS) were examined. Mean CIPA prior to the onset of STCS was elevated above normal values and fell significantly to previously established normal values following the seizure. CPS were not associated with elevated mean CIPA pre- or post-seizure. STCS were associated with a reduction in anti convulsant dosage and with elevation of pre-ictal CIPA. We propose that elevation of cardiac parasympathetic activity pre-ictally may be a marker for secondary generalisation of seizures. PMID- 10372562 TI - MR imaging of implanted depth and subdural electrodes: is it safe? AB - This study evaluates the safety of imaging chronic epilepsy patients with intracranial depth and subdural electrodes by magnetic resonance (MR). To identify an epileptogenic focus, the precise location of the electrode contacts is necessary, and MR can provide this information. However, many neurosurgeons and neuroradiologists are hesitant to scan patients by MR with these implanted, metallic electrodes for fear of electrode displacement, current induction or heating secondary to the strong magnetic field. In the present study, the subdural electrodes were made of stainless steel with either stainless steel or platinum contacts. The depth electrodes were made of either platinum or a nickel chromium alloy (nichrome). We reviewed 98 cases in which patients with implanted depth electrodes, subdural electrodes, or both underwent MR scanning. A total of 143 depth electrodes, 688 subdural strips, and 38 subdural grids were implanted in the 98 procedures. MR scanning was performed on a 1.5-T unit and consisted of T1, T2, and/or spoiled gradient echo pulse sequences. There were no documented complications related to the MR scans. Based on this study and a review of the literature, we feel that MR imaging can safely localize intracranial electrodes. PMID- 10372563 TI - Characterization of the K+ channel opening effect of the anticonvulsant retigabine in PC12 cells. AB - Retigabine (D-23129) is a new anticonvulsant compound which acts as a K+ channel opener in neuronal cells. The aim of the present study was to further characterize the retigabine induced K+ current. In nerve growth factor treated PC12 cells and in rat cortical neurones the application of retigabine activated a K+ current. In contrast, however, no K+ current activation was observed in untreated PC12 and in glial cells which were cultivated together with the neuronal cells. To characterise the retigabine activated K+ current, K+ channel blockers were used. The retigabine induced current was not affected by 1 and 10 mM 4-aminopyridine (4AP). Ba2+ 1 mM resulted in a reduction of 88.6+/-3.0% (n = 5); 10 mM abolished the current. Tetraetylamonium (TEA), 1 and 10 mM, reduced the current by 23.6+/-3.1 and 61.6+/-3.7%, respectively. To investigate the current/voltage (I/V) relation of the current initiated by retigabine (10 microM), cells were clamped to a holding potential of -80 mV and a ramp stimulation protocol (-120 to +60 mV in 5 s) was applied prior to and during application of retigabine. Subtraction of the two traces yielded the current induced by retigabine. A nearly linear relationship was determined between - 120 and -40 mV. At potentials positive to - 40 mV, the response was variable. This was due to the additionally observed weak blocking effect of retigabine on delayed rectifier (Kdr) currents. If the ramp was applied in the presence of 10 mM 4AP to block Kdr, a nearly linear I/V-relationship was present from -120 to +60 mV. The comparison of the I/V relation and pharmacology with published K+ channel subtypes gives evidence that an unknown neuronal K+ channel subtype may be involved. PMID- 10372565 TI - Rectal diazepam: pitfalls of excessive use in refractory epilepsy. AB - INTRODUCTION: The rectal administration of diazepam (DZP) has provided a safer existence to many epilepsy patients and their carers, when prolonged or serial seizures are present. However, in patients with frequent seizures, chronic, intermittent over-administration may occur, particularly in the presence of multihandicaps. METHODS: Six patients who experienced untoward effects from excessive rectal DZP are reported. In two patients, serial plasma levels of DZP and its active metabolites were monitored. RESULTS: Three patients exhibited a pattern of cyclic reappearance of seizures, interrupted by rectal DZP, followed by sedation and gradual awakening. The intervals were approximately 4 days. The three other patients had variable and complex symptoms with serial seizures and alternating states of tension, apathy, and sleepiness. The plasma levels of DZP and desmethyl-DZP showed rapid fluctuations. CONCLUSION: When rectal DZP is prescribed, chronic and excessive administration should be avoided. Fluctuating plasma-levels may probably support a cyclic reappearance of seizures in some patients. The combination of high bolus doses and a rapid drug clearance due to enzyme inducing co-medication may probably increase the risk for rebound reactions. Toxic, withdrawal, and epileptic symptoms may be intermingled and difficult to manage. A replacement strategy in the form of a prophylactic, oral, low dose benzodiaepine regimen may facilitate the discontinuation of this prescription pattern. Adequate counselling and medically appropriate, written directions for the use of rectal DZP is mandatory. PMID- 10372566 TI - Hippocampal alterations of apolipoprotein E and D mRNA levels in vivo and in vitro following kainate excitotoxicity. AB - Alteration in the expression of apolipoprotein E (ApoE) and apolipoprotein D (ApoD) genes was evaluated in rat, 7 days following status epilepticus (SE) induced by intra-amygdala injection of kainate (KA), and in organotypic hippocampal cultures, 2 days after a single 1 h exposure to KA. Global polyadenylated RNA (poly A+) steady state, assessing global regulation of mRNA transcription was first measured in cortices and hippocampi from each animal and in the organotypic cultures. No alteration due to KA treatment was observed and individual concentrations of ApoE and ApoD mRNA species were therefore measured and comparative analysis performed. In the cortices of KA-treated animals, ApoE and ApoD mRNA levels did not show statistically significant changes. In contrast, in hippocampi, 7 days after SE, ApoE and ApoD mRNA levels were significantly increased, respectively, by 123 and 138%. This in vivo effect was confirmed in vitro on organotypic cultures, where KA treatment increased ApoE and ApoD mRNA expressions, respectively, by 72 and 61%. These observations indicate that lipidic metabolism is modified in the lesioned structure and suggest an increased traffic of lipids and a need for more ApoE and D in the hippocampus during the period of recovery and restructuration that follows severe seizures. PMID- 10372564 TI - Cognitive abilities and adjustment with gabapentin: results of a multisite study. AB - The cognitive and quality of life effects of gabapentin are not yet well explored. While preliminary work in the area has provided positive findings, a large double-blinded study has been needed to explore this area more thoroughly. From 24 sites in North America, 201 adults were studied who had uncontrolled complex partial seizures with or without secondary generalization. Attempts were made to convert each patient from one or two marketed antiepileptic drugs (AEDs) taken in baseline to gabapentin monotherapy (600, 1200, or 2400 mg/day). Tests of cognitive abilities and adjustment were administered at the end of the 8-week baseline period and at the end of the 26-week double-blind treatment period. Analyses of baseline to treatment period changes were conducted for each dose group in comparison with a reference group of placebo-treated patients from another study. In the area of cognitive functioning, no changes in any of the gabapentin groups were found in comparison with the reference group. In the area of adjustment and mood, however, improvement with gabapentin administration was noted on several variables pertaining to emotional and interpersonal adjustment. These results are consistent with findings from previous studies. PMID- 10372568 TI - Scopolamine-induced convulsions in food given fasted mice: effects of clonidine and tizanidine. AB - We recently reported that scopolamine pretreated mice fasted for 48 h developed clonic convulsions soon after they were allowed to eat ad libidum. Pretreatment with MK-801, the non-competitive NMDA antagonist, decreased the incidence of these convulsions. We suggested that a possible scopolamine-induced glutamatergic hyperactivity could account for these convulsions. Using alpha2-agonists, clonidine, which has been shown to inhibit glutamate release, and tizanidine, the present study was performed to find some additional data for the role of glutamate in the underlying mechanism of scopolamine-induced convulsions in food given fasted mice. Animals fasted for 48 h and pretreated (i.p.) with saline, clonidine (0.05, 0.10, 1 mg/kg) or tizanidine (0.10, 0.15, 0.30, 0.45 mg/kg) were treated (i.p.) with either saline or scopolamine (3 mg/kg). Then 20 min later, they were allowed to eat ad libidum and were observed for 30 min for the incidence and onset of clonic convulsions. All doses of clonidine pretreatment completely suppressed (0%) scopolamine-induced clonic convulsions (75%). On the other hand, only 0.15 mg/kg tizanidine pretreatment significantly decreased (15%) the incidence of convulsions; however as well as 0.15 mg/kg, both 0.30 and 0.45 mg/kg tizanidine pretreatments significantly increased latency to the onset of convulsions. PMID- 10372567 TI - Safety of rapid intravenous infusion of valproate loading doses in epilepsy patients. AB - IV valproate may be given to patients requiring rapid elevation of serum valproate or for patients unable to take oral medication. Previously we established the loading dose of IV valproate needed to achieve serum concentrations of > 100 ug/ml. This study evaluated the safety of rapid infusion of valproate to achieve high therapeutic levels. Twenty-four infusions of IV valproate were carried out electively in twenty-one patients with epilepsy (ages 2-54 years). The dose ranged from 21-28 mg/kg (mean 24.2 mg/kg). Target infusion rates were 3 or 6 mg/kg per min, yielding an infusion duration of 4.17 or 8.34 min. ECG was monitored; blood pressure was measured before and after infusion. Post infusion serum valproate concentrations were 64-204.1 ug/ml (mean 132.6). There were no significant changes in blood pressure and no ECG abnormalities observed. Transient pain occurred at the site of injection in five patients, associated with redness in two. This appeared to related to the concentration of valproic acid in the infusion fluid. We conclude that a loading dose of IV valproate can be administered safely and rapidly. This finding enables further studies evaluating IV valproate as a non-sedative anticonvulsant in the management of status epilepticus. PMID- 10372569 TI - Heterogeneous distribution of polyamines in temporal lobe epilepsy. AB - Polyamine contents were determined in human temporal lobe epilepsy. In the seven patients studied, stereoelectroencephalography (SEEG) located the epileptogenic focus in Ammon's horn and neuropathological findings were limited to hippocampal gliosis and sclerosis. Each polyamine exhibited a specific regional distribution. The most important variations were observed for spermidine and spermine while putrescine levels varied less. The regional variation was predominant in middle > posterior > anterior parts of the temporal lobe. Spermine contents and the spermidine/spermine (SPD/SPM) index varied especially in the middle and posterior parts of the hippocampus. Metabolic SPD/SPM index and spermidine levels were found to be drastically increased in almost all limbic parts when compared to neocortical regions. The opposite was observed for spermine. The heterogeneous distribution of polyamines was compared to abnormal electrical activities recorded by SEEG: SPD/SPM index and spermidine levels were sharply increased in seizure onset areas and high levels of spermine were detected in temporal cortex propagation areas. The presently reported heterogeneity of polyamine contents might contribute to modulate differentially the local control of excitability in human temporal epilepsy. PMID- 10372570 TI - Localization of brain reinforcement mechanisms: intracranial self-administration and intracranial place-conditioning studies. AB - Intracranial self-administration (ICSA) and intracranial place conditioning (ICPC) methodologies have been mainly used to study drug reward mechanisms, but they have also been applied toward examining brain reward mechanisms. ICSA studies in rodents have established that the ventral tegmental area (VTA) is a site supporting morphine and ethanol reinforcement. ICPC studies confirmed that injection of morphine into the VTA produces conditioned place preference (CPP). Further confirmation that activation of opioid receptors within the VTA is reinforcing comes from the findings that the endogenous opioid peptide met enkephalin injected into the VTA produces CPP, and that the mu- and delta-opioid agonists, DAMGO and DPDPE, are self-infused into the VTA. Activation of the VTA dopamine (DA) system may produce reinforcing effects in general because (a) neurotensin is self-administered into the VTA, and injection of neurotensin into the VTA produces CPP and enhances DA release in the nucleus accumbens (NAC), and (b) GABA(A) antagonists are self-administered into the anterior VTA and injections of GABA(A) antagonists into the anterior VTA enhance DA release in the NAC. The NAC also appears to have a major role in brain reward mechanisms, whereas most data from ICSA and ICPC studies do not support an involvement of the caudate-putamen in reinforcement processes. Rodents will self-infuse a variety of drugs of abuse (e.g. amphetamine, morphine, phencyclidine and cocaine) into the NAC, and this occurs primarily in the shell region. ICPC studies also indicate that injection of amphetamine into the shell portion of the NAC produces CPP. Activation of the DA system within the shell subregion of the NAC appears to play a key role in brain reward mechanisms. Rats will ICSA the DA uptake blocker, nomifensine, into the NAC shell; co-infusion with a D2 antagonist can block this behavior. In addition, rats will self-administer a mixture of a D1 plus a D2 agonist into the shell, but not the core, region of the NAC. The ICSA of this mixture can be blocked with the co-infusion of either a D1 or a D2 antagonist. However, the interactions of other transmitter systems within the NAC may also play key roles because NMDA antagonists and the muscarinic agonist carbachol are self-infused into the NAC. The medial prefrontal (MPF) cortex supports the ICSA of cocaine and phencyclidine. The DA system also seems to play a role in this behavior since cocaine self-infusion into the MPF cortex can be blocked by co infusing a D2 antagonist, or with 6-OHDA lesions of the MPF cortex. Limited studies have been conducted on other CNS regions to elucidate their role in brain and drug reward mechanisms using ICSA or ICPC procedures. Among these regions, ICPC findings suggest that cocaine and amphetamine are rewarding in the rostral ventral pallidum (VP); ICSA and ICPC studies indicate that morphine is rewarding in the dorsal hippocampus, central gray and lateral hypothalamus. Finally, substance P mediated systems within the caudal VP (nucleus basalis magnocellularis) and serotonin systems of the dorsal and median raphe nuclei may also be important anatomical components involved in brain reward mechanisms. Overall, the ICSA and ICPC studies indicate that there are a number of receptors, neuronal pathways, and discrete CNS sites involved in brain reward mechanisms. PMID- 10372571 TI - The effects of high fat diets and environmental influences on cognitive performance in rats. AB - As part of a continuing investigation of the relationship between dietary factors and cognitive function, the present study examined the combined effects of environmental influences and high-fat diets on learning and memory. Following 3 months of dietary (20% by weight fat diets, composed primarily of either beef tallow or soybean oil versus standard laboratory chow) and environmental treatments (standard, enriched or impoverished), subjects were tested on a variable interval delayed alternation (VIDA) task which measures learning and memory functions that differentially involve specific brain regions. The results confirmed the negative effects of high fat diets, relative to chow, on all aspects of VIDA performance and showed that environmental enrichment overcame deficits associated with dietary fat. Housing rats fed high-fat diets in an impoverished environment did not further exacerbate cognitive deficits observed in such rats living under standard conditions. By comparison, chow-fed rats exhibited no benefit associated with the enriched environment on any aspect of task performance, and only a transitory learning impairment when housed in an impoverished environment. The results show that high fat diets and environmental conditions influence cognitive function and that these two factors interact with one another to produce different profiles of benefits and impairments. PMID- 10372572 TI - Behavioural and hormonal differences between two Lewis rat lines. AB - Lewis (LEW) is an inbred strain of rats frequently used as an animal model of autoimmune diseases. However, there is evidence that some lines of LEW rats develop autoimmune diseases more readily than do other LEW rat lines. Because the hypothalamus-pituitary-adrenal system is involved in the pathophysiology of these diseases, we compared two LEW lines (SsNHsd and HANRijHsd) in their behavioural and neuroendocrine response to stress. In addition, we studied the psychostimulant effects of acute and repeated amphetamine in these two LEW rat lines. HAN rats were less active in the open field test and showed faster habituation of novelty-induced locomotion. The acoustic startle response was lower in HAN than in SSN rats, whereas prepulse inhibition of the startle response was greater in the HAN than in the SSN LEW subline. Moreover, HAN rats showed impaired acquisition of the two-way active avoidance response relative to SSN rats. The psychostimulant effects of acute amphetamine were smaller in HAN rats. Following repeated injections of amphetamine, behavioural sensitization to the psychostimulant effects of amphetamine was more pronounced in HAN than in SSN rats. Basal concentrations of serum corticosterone did not differ between the two rat lines. Following stress, however, HAN rats showed slightly higher corticosterone secretion than SSN rats. Our results show that two sublines of the LEW inbred strain of rats show profound behavioural differences which are only marginally paralleled by differences at the level of the HPA system. PMID- 10372573 TI - Is digital dexterity really related to corticospinal projections?: a re-analysis of the Heffner and Masterton data set using modern comparative statistics. AB - Using a data set of 69 different mammalian species, Heffner and Masterton propose that the longer and deeper the fibres of the corticospinal tract, the greater an animal's digital dexterity. Because of the effects that phylogeny may have upon the extant phenotype of a given species, however, data from a wide range of species can rarely be considered to represent fully independent data points. Using modern comparative statistics, which incorporate phylogenetic information, we reanalysed their data set such that the assumption of independence was not violated. If Heffner and Masterton's hypothesis is correct, then one would expect evidence of strong correlated evolution between corticospinal tract anatomy and digital dexterity once the effects of the phylogenetic relationships between the species in the data set have been removed. The results show that a distinct bias in the number of primate species sampled by Heffner and Masterton significantly affected their findings. Furthermore, once phylogeny has been taken into account, only the length of the corticospinal tract fibres showed a significant relationship with two out of the four behaviours analysed, digital dexterity and hand-eye coordination. Based upon our results we recommend the use of modern comparative statistics for synthesising neural structure and behaviour, rather than examining structure function relationships in an ahistorical context. It is also evident that there is a need for data on the length and depth of the corticospinal fibres for a greater range of species so that the relationship between the corticospinal tract structure and motor behaviour for mammals as a whole can be more readily interpreted. PMID- 10372575 TI - Reinforcement enhances hippocampal acetylcholine release in rats: an in vivo microdialysis study. AB - Rats were trained to press a lever under a 'Multiple FI-60s and Extinction' schedule with food reinforcements. After learning the task, an in vivo microdialysis probe was inserted into the dentate gyrus and CA3 regions of the hippocampus, and the sequential changes in the dialysate acetylcholine (ACh) concentration were analyzed. In the session, two fixed-interval of reinforcement (FI) components (for 20 min) and two extinction (EXT) components (for 30 min) were alternated to examine the correlation between behavioral and neurochemical outcomes. The dialysate ACh concentration increased during the FI component and returned to the baseline during the EXT component of the schedule. Next, in order to dissociate the effect of discrimination from the effect of rewarding on the neurochemical changes in the hippocampus, we used a final TEST period (for 20 min) during which the actual schedule was extinction but the discriminative stimulus was on, i.e. the manifest condition of the test period was reinforcement. In the TEST period, the animals pressed the lever with almost the same frequency as during the FI component; however, the dialysate ACh concentration did not increase above the baseline concentration. These results suggest that ACh release in the rat hippocampus is associated with reinforcement but not with discrimination in operant conditioning. PMID- 10372574 TI - Issues in the pharmacological modification of cocaine conditioning: evidence that the stimulus properties of drugs can interact with contextual cues to activate or inactivate cocaine conditioned stimuli. AB - Cocaine conditioned stimuli are capable of eliciting cocaine craving in individuals with a history of cocaine use. As a consequence, there have been a number of attempts using animal models to identify pharmacological treatments which can attenuate cocaine conditioned effects. The emphasis in these studies has been to employ drug doses which do not have response effects that could directly alter the conditioned drug response. A drug treatment may not have a response effect but still have drug stimulus effects which could interact with and modify the cocaine conditioned stimulus. In order to experimentally investigate this important issue, two experiments are reported. In one experiment, rats were co-administered 0.1 mg/kg MK-801 either with cocaine (10 mg/kg) or with saline; in the other experiment 3.0 mg/kg buspirone was co administered with either cocaine (10 mg/kg) or with saline. The MK-801 and buspirone treatments did not affect spontaneous activity levels or alter the unconditioned cocaine stimulant effect. In tests for conditioning, however, the effects of buspirone and MK-801 depended upon their association with cocaine. If MK-801 and buspirone had no association with cocaine then these drugs inactivated the cocaine conditioned stimulant response. If MK-801 and buspirone had been co administered with cocaine, then, in saline conditioning tests, no cocaine conditioning was observed. If the conditioning tests were conducted following MK 801 or buspirone treatment, however, cocaine conditioning was elicited. Altogether, these studies demonstrate that the stimulus properties of drugs can interact with contextual stimuli to inactivate or activate cocaine conditioned stimuli. In the search for drugs which may prevent cocaine craving, therefore, the stimulus properties of drugs provide an important mechanism for the modification of cocaine conditioned stimuli. PMID- 10372576 TI - Task-specific enhancement of short-term, but not long-term, memory by class I metabotropic glutamate receptor antagonist 1-aminoindan-1,5-dicarboxylic acid in rats. AB - Pharmacological application of broad agonists and antagonists has supported the notion of a potential role of metabotropic glutamate receptors (mGluRs) in learning and memory formation, but the specific function of the different classes or individual subtypes remains elusive. Furthermore, our knowledge with respect to different learning mechanisms is still fragmentary. In an attempt to clarify further the function of mGluRs in learning, rats were trained in various paradigms in the presence/absence of the specific class I antagonist 1-aminoindan 1,5-dicarboxylic acid (AIDA). Intraperitoneal application of AIDA prior to training led to enhanced within-session performance in animals trained in a positively reinforced reference memory task in a three-choice maze. However, this enhancement did not result in increased retention as measured by the number of correct responses during the first four trials of each session on subsequent days. The increase was purely an enhancement in within-session performance, required doses higher than 2 mg/kg, and was not accompanied by an unspecific increase in activity as monitored in the open field. By contrast, AIDA animals trained in a combined shock-reinforced contextual and cue conditioning paradigm demonstrated a pronounced retention deficit compared with controls in conditioning to the context, but not the cue (a high-frequency tone). Although within-session performance during context and cue periods was slightly increased in the AIDA group, the difference did not reach significance. Drug-induced hyperactivity, which could account for the memory deficit, was excluded by recordings of activity in specific activity cages. These results shed new light on the possible function of class I mGluRs in learning and memory formation and imply that systemic blockade of class I mGluRs may enhance short-term memory under certain learning conditions. PMID- 10372577 TI - Management of unruptured intracranial aneurysm in Japan: a Markovian decision analysis with utility measurements based on the Glasgow Outcome Scale. AB - The purpose of this study was to analyze the management of individual patients with unruptured intracranial aneurysms (UN-ANs) using a decision-analytic approach. Transition probabilities among Glasgow Outcome Scale (GOS) categories were estimated from the published literature and data from patients who had been treated at Kitasato University Hospital. Utilities were obtained from 140 health providers based principally on the GOS. Baseline analysis for a healthy 40-year old man with an anterior UN-AN less than 10 mm in diameter showed that the quality-adjusted life expectancies for preventive operation and follow-up were 15.34 and 14.66 years, respectively. For a follow-up strategy to be preferred, the annual rupture rate had to be as low as 0.9%. These results were sustained through extensive sensitivity analysis. The results support preventive operation for UN-ANs, and identify problems that can be clarified with a well-designed stratified clinical trial. PMID- 10372579 TI - The danger of applying group-level utilities in decision analyses of the treatment of localized prostate cancer in individual patients. AB - The optimal management strategy for men who have localized prostate cancer remains controversial. This study examines the extent to which suggested treatment based on the perspective of a group or society agrees with that derived from individual patients' preferences. A previously published decision analysis for localized prostate cancer was used to suggest the treatment that maximized quality-adjusted life expectancy. Two treatment recommendations were obtained for each patient: the first (group-level) was derived using the mean utilities of the cohort; the second (individual-level) used his own set of utilities. Group-level utilities misrepresented 25-48% of individuals' preferences depending on the grade of tumor modeled. The best kappa measure achieved between group and individual preferences was 0.11. The average quality-adjusted life years lost due to misrepresentation of preference was as high as 1.7 quality-adjusted life years. Use of aggregated utilities in a group-level decision analysis can ignore the substantial variability at the individual level. Caution is needed when applying a group-level recommendation to the treatment of localized prostate cancer in an individual patient. PMID- 10372578 TI - Testing for the BRCA1 and BRCA2 breast-ovarian cancer susceptibility genes: a decision analysis. AB - OBJECTIVE: The authors developed a Markov decision model to evaluate the health implications of testing for mutations in the BRCA1 and BRCA2 breast-ovarian cancer susceptibility genes. Prophylactic measures considered included various combinations of immediate and delayed bilateral mastectomy and oophorectomy or taking no action. METHODS: The model incorporated the likelihood of developing breast and/or ovarian cancer, survival, and quality of life. Parameter values were taken from public databases, the published literature, and a survey of cancer experts. Outcomes considered were additional life expectancy and quality adjusted life years (QALYs). Results are reported for 30-year-old cancer-free women at various levels of hereditary risk. RESULTS AND CONCLUSIONS: The vast majority of women will not benefit from testing because their pre-test risks are low and surgical prophylaxis is undesirable. However, women who have family histories of early breast and/or ovarian cancer may gain up to 2 QALYs by allowing genetic testing to inform their decisions. PMID- 10372580 TI - Estimating general-population utilities using one binary-gamble question per respondent. AB - This study used a single binary-gamble question per health state per respondent to obtain societal preferences for the health states intermittent claudication and major amputation and compare those with Health Utilities Indices obtained from patients, to test the feasibility of this method, and to investigate whether the utility depends on the presentation of a vignette as generic vs disease specific. A random sample of the general U.S. population (n = 1,003) was randomly divided into ten subgroups. In telephone interviews, subjects answered one binary gamble question in a standard-gamble format for each of two health states. The risks of death varied across subgroups but not between health states. Mean utility was estimated by the area above the proportional distribution of responses indicating acceptance of the gamble. The method is based on the binary choice method used in contingent-valuation studies of willingness to pay. The health states were alternatively described by generic and disease-specific vignettes in two subsamples. The results suggest that the binary-gamble question can be used to obtain societal preferences for health states, and that disease specific descriptions yield lower utilities compared with generic descriptions of health states. PMID- 10372581 TI - Patients' utilities for cancer treatments: a study of the chained procedure for the standard gamble and time tradeoff. AB - OBJECTIVE: Temporary health states cannot be measured in the traditional way by means of techniques such as the time tradeoff (TTO) and the standard gamble (SG), where health states are chronic and are followed by death. Chained methods have been developed to solve this problem. This study assesses the feasibility of a chained TTO and a chained SG, and the consistency and concordance between the two methods. PATIENTS AND METHODS: Seventy female early-stage breast cancer patients were interviewed. In using both chained methods, the temporary health state to be evaluated was weighed indirectly with the aid of a temporary anchor health state. The patients were asked to evaluate their actual health states, a hypothetical radiotherapy scenario, and a hypothetical chemotherapy scenario. RESULTS: Sixty eight patients completed the interview. The use of the anchor health state yielded some problems. A significant difference between the means of the TTO and the SG was found for the anchor health state only. For the other health states, the results were remarkably close, because the design avoided some of the bias effects in traditional measurements. CONCLUSION: The feasibility and the consistency of the chained procedure were satisfactory for both methods. The problems regarding the anchor health state can be solved by adapting the methods and by the use of a carefully chosen anchor health state. The chained method avoids biases present in the conventional method, and thereby the TTO and the SG may be reconciled. Moreover, there are several psychological advantages to the method, which makes it useful for diseases with uncertain prognoses. PMID- 10372582 TI - Time-tradeoff values and standard-gamble utilities assessed during telephone interviews versus face-to-face interviews. AB - The purpose of this study was to compare time-tradeoff values and standard-gamble utilities obtained during telephone interviews with those obtained through face to-face interviews. Sixty-five patients with peripheral arterial occlusive disease completed both interviews. One week prior to the telephone interview, the patients received by mail a questionnaire in which the value and utility measures were presented in writing. The face-to-face interviews used the same questions, but the interviewer used visual aids. The mean time-tradeoff values were 0.84 (SD 0.20) vs 0.86 (SD 0.17) for the telephone and face-to-face interviews, respectively (p = 0.31). The mean standard-gamble utilities were 0.93 (SD 0.16) vs 0.92 (SD 0.17) for the telephone and face-to-face interviews, respectively (p = 0.26). In conclusion, telephone interviews yield similar time-tradeoff values and standard-gamble utilities compared with face-to-face interviews, suggesting that telephone interviews can replace face-to-face interviews. PMID- 10372583 TI - Attitudes of obstetricians and gynecologists toward hormone replacement therapy. AB - OBJECTIVE: To determine the attitudes of obstetricians and gynecologists toward hormone replacement therapy (HRT), and the beliefs and intuitions that affected those attitudes. DESIGN: A questionnaire was sent to 1,000 gynecologists in the United States; 328 replies were received. The questionnaire asked about effects of HRT, practices concerning HRT, and decisions in hypothetical scenarios. RESULTS: The respondents strongly favored HRT, and they were well informed about its effects on osteoporosis, cardiovascular disease, and breast cancer. They were aware of conflicting findings concerning breast cancer. The strength of their recommendation of HRT was sensitive to patient differences in risk factors. The respondents also showed four biases hypothesized to cause resistance to HRT: omission bias (more concern about harmful acts than harmful omissions); proportionality bias (attention to relative risk rather than risk differences); naturalness bias (preference for the natural); and ambiguity (avoiding options with missing information). Proportion bias, naturalness bias, and (weakly) omission bias were related to less favorable attitudes toward HRT. CONCLUSION: Although specialists are highly favorable toward HRT in general, some negativity toward HRT may result from decision biases. PMID- 10372584 TI - Clinical diagnosis and the order of information. AB - BACKGROUND: Information order can influence judgment. However, it remains unclear whether the order of clinical data affects physicians' interpretations of these data when they are engaged in familiar diagnostic tasks. METHODS: Of 400 randomly selected family physicians who were given a questionnaire involving a brief written scenario about a young woman with acute dysuria, 315 (79%) returned usable responses. The physicians had been randomized into two groups, and both groups had received the same clinical information but in different orders. After learning the patient's chief complaint, physicians received either the patient's history and physical examination results followed by the laboratory data (the H&P first group) or the laboratory data followed by the history and physical examination results (the H&P-last group). The results of the history and physical examination were supportive of the diagnosis of UTI, while the laboratory data were not. All physicians judged the probability of a urinary tract infection (UTI) after each piece of information. RESULTS: The two groups had similar mean estimates of the probability of a UTI after learning the chief complaint (67.4% vs 67.8%, p = 0.85). At the end of the scenario, the H&P-first group judged UTI to be less likely than did the H&P-last group (50.9% vs 59.1%, p = 0.03) despite having identical information. Comparison of the mean likelihood ratios attributed to the clinical information showed that the H&P-first group gave less weight to the history and physical than did the H&P-last group (p = 0.04). CONCLUSIONS: The order in which clinical information was presented influenced physicians' estimates of the probability of disease. The clinical history and physical examination were given more weight by physicians who received this information last. PMID- 10372585 TI - Tests of preferential independence in the QALY model. AB - The author presents evidence for the descriptive adequacy of the quality-adjusted life years (QALY) model as applied to health profiles. One important assumption of the model is preferential independence: if two profiles have the same health state during year X, then preference between them does not switch if the level of health changes during year X. In experiment 1, 30-year health profiles were used to perform 27 empirical tests of independence with 98 subjects. Independence was reliably satisfied in all 27 tests. In experiment 2, 15 additional tests were conducted. These tests had been specifically designed to be more sensitive to independence violations, but independence was still mostly satisfied. In both experiments, the conclusions about independence hold regardless of what discount rate is used. These results act as a "lower bound" on the validity of the QALY model for health profiles. PMID- 10372586 TI - Ambulatory blood pressure monitoring and diagnostic errors in hypertension: a Bayesian approach. AB - Random variability of blood pressure complicates the diagnosis and subsequent treatment of hypertension. To evaluate the importance of the number of blood pressure measurements in the correct diagnosis and control of hypertension, the authors used a Bayesian model to estimate the true average blood pressure of a group of newly diagnosed hypertensives, then calculated the diagnostic error that would result from monitoring methods using 24 daytime measurements or from using only three random monitoring measurements. The study population consisted of 129 individuals with newly diagnosed mild hypertension according to standard criteria, who were also evaluated with an ambulatory blood pressure monitor. In true normotensives (daytime diastolic blood pressure <90 mm Hg), the negative predictive value with three measurements was 0.92, and it rose to 0.96 with monitoring methods. In mild hypertensives (90-104 mm Hg), the positive predictive value was 0.64 with three measurements and 0.84 with monitoring methods, thus reducing the rate of false mild hypertensives from 35% to 15%. Finally, in patients with moderate or severe hypertension (>104 mm Hg), the positive predictive value improved from 0.26 with three readings to 0.61 with monitoring methods. Similar results were observed with daytime systolic pressure measurements. As the number of measurements increased, the diagnostic error due to the random variability of blood pressure became progressively smaller. In cases of hypertension, the large improvement in predictive values may justify using monitoring methods to confirm standard diagnosis. PMID- 10372588 TI - Neural networks, logistic regression, and calibration: a reply. PMID- 10372587 TI - A Bayesian approach to a general regression model for ROC curves. AB - A fully Bayesian approach to a general nonlinear ordinal regression model for ROC curve analysis is presented. Samples from the marginal posterior distributions of the model parameters are obtained by a Markov-chain Monte Carlo (MCMC) technique- Gibbs sampling. These samples facilitate the calculation of point estimates and credible regions as well as inferences for the associated areas under the ROC curves. The analysis of an example using freely available software shows that the use of noninformative vague prior distributions for all model parameters yields posterior summary statistics very similar to the conventional maximum-likelihood estimates. Clinically important advantages of this Bayesian approach are: the possible inclusion of prior knowledge and beliefs into the ROC analysis (via the prior distributions), the possible calculation of the posterior predictive distribution of a future patient outcome, and the potential to address questions such as: "What is the probability that a certain diagnostic test is better in one setting than in another?" PMID- 10372589 TI - Clarifying the management decision for early breast cancer. PMID- 10372590 TI - Law and ethics (future risk of cancer). PMID- 10372591 TI - Judgmental psychology. Contingent valuation. PMID- 10372592 TI - In vivo and ex vivo uptake of albendazole and its sulphoxide metabolite by cestode parasites: relationship with their kinetic behaviour in sheep. AB - The current experiments correlate the disposition kinetics of albendazole (ABZ) following its intravenous (i.v.) and intraruminal (i.r.) administrations to Moniezia spp.-infected sheep, with the pattern of drug/metabolite uptake by tapeworms collected from treated animals. The ex vivo uptake pattern of ABZ and albendazole sulphoxide (ABZSO) by the same cestode parasite was also investigated. Naturally infected (Moniezia spp.) Corriedale lambs were treated with ABZ by either i.v. (Group A, n = 15) or i.r. (Group B, n = 15) administration at 7.5 mg/kg. Plasma and abomasal fluid samples were obtained over a 120-h period. Two animals per group were killed at 0.5, 1, 2, 4 and 6 h post treatment; parasite material (tapeworms), bile and intestinal fluid samples were recovered. Furthermore, Moniezia spp. tapeworms obtained from sheep killed at the local abattoir were incubated with either ABZ or ABZSO for different time periods in a Kreb's Ringer Tris buffer (ex vivo experiments). Samples were analysed by high performance liquid chromatography for ABZ, ABZSO and albendazole sulphone (ABZSO2). ABZ plasma concentrations decreased rapidly and were not detectable beyond 10 h following i.v. administration. ABZSO and ABZSO2 were the metabolites recovered in plasma after both treatments. ABZ and its metabolites were extensively distributed to the digestive tract, mainly into the abomasal fluid, after the i.v. and i.r. administrations. The parent drug and its active ABZSO metabolite were recovered in tapeworms collected from both i.v. and i.r. treated lambs. However, the availability of both ABZ and ABZSO was higher in parasite material recovered from i.v. treated animals. The uptake of ABZ by the cestode parasite, both in vivo and ex vivo, was significantly greater than that of its sulphoxide metabolite, which agrees with the higher lipophilicity of the parent drug. PMID- 10372593 TI - The suppressive effects of lipopolysaccharide-induced acute phase response on hepatic cytochrome P450-dependent drug metabolism in rabbits. AB - The acute phase response (APR) was induced by five separate intravenous (i.v.) injections of Escherichia coli lipopolysaccharide (LPS, 17 microg/kg each time) in rabbits, with intervals of 1 h. This model was used to study the effects of APR on the activities of hepatic microsomal cytochrome P450 (CYP)-dependent enzyme including drug metabolism. Five female rabbits were included in each of four groups, a control group and three LPS-treated groups (group I, II and III). The rabbits of the control, group I, II and III were killed at 1, 1, 3 and 7 days after saline (control only) or the LPS injection, respectively. The APR was confirmed by increases in rectal body temperature, plasma concentrations of interleukin-6 and C-reactive protein (CRP). Pharmacokinetics of antipyrine before death were examined in every group. Antipyrine was administered (5 mg/kg) at 24 h (control and group I), 3 days (group II) and 7 days (group III) after the first LPS injection. Total body clearance (Cl(tot)) of antipyrine tended to decrease in group I. All the livers were excised for measuring CYP-dependent activities. Total CYP content and several CYP-dependent activities (aminopyrine N demethylation, aniline 4-hydroxylation and caffeine 3-demethylation) decreased in group I. The maximum velocity (Vmax) values of those enzymes, and the amount of CYP1A1/1A2 and CYP2E1 apoproteins appeared to decrease. Michaelis constant (Km) values of those enzymes were not affected by the APR. Rectal body temperature recovered to normal at 3 days after the first LPS injection in group II and III. The concentration of CRP, albumin, total CYP content and the plasma clearance of antipyrine returned to the control levels at 7 days after the first LPS injection. These results suggest that the metabolism of drugs, including CYP dependent drug metabolizing activity, is suppressed markedly in incipient APR induction in rabbits, and the drug metabolizing capacity is returned to normal at 7 days after APR induction. PMID- 10372594 TI - A pharmacodynamic and pharmacokinetic study with vedaprofen in an equine model of acute nonimmune inflammation. AB - The pharmacodynamics and enantioselective pharmacokinetics of vedaprofen were studied in six ponies in a two period cross-over study, in which a mild acute inflammatory reaction was induced by carrageenan soaked sponges implanted subcutaneously in the neck. Vedaprofen, administered intravenously at a dosage of 1 mg/kg, produced significant and prolonged inhibition of ex vivo serum thromboxane B2 (TXB2) synthesis and short-lived inhibition of exudate prostaglandin E2 (PGE2) and TXB2 synthesis. Vedaprofen also partially inhibited oedematous swelling and leucocyte infiltration into exudate. Vedaprofen displayed enantioselective pharmacokinetics, plasma concentrations of the R(-) enantiomer exceeding those of S(+) vedaprofen. The plasma concentration ratio, R:S, increased from 69:31 at 5 min to 96:4 at 3 h and plasma mean AUC values were 7524 and 1639 ng x h/mL, respectively. Volume of distribution was greater for S(+) vedaprofen, whilst elimination half-life (t(1/2beta)) and mean residence time were greater for R(-) vedaprofen. The penetration of vedaprofen into inflammatory exudate was also enantioselective. For R(-) and S(+) vedaprofen maximum concentration (Cmax) values were 2950 and 1534 ng/mL, respectively, and corresponding AUC values were 9755 and 4400 ng x h/mL. Vedaprofen was highly protein bound (greater than 99%) in both plasma and exudate. The significance of these data for the therapeutic use of vedaprofen is discussed. PMID- 10372595 TI - Mepivacaine: its pharmacological effects and their relationship to analytical findings in the horse. AB - Mepivacaine is a local anaesthetic drug that is widely used in equine medicine and is classified by the Association of Racing Commissioners International (ARCI) as a Class 2 foreign substance that may cause regulators to impose significant penalties if residues are identified in post-race urine samples. Therefore, an analytical/pharmacological database was developed for this agent and its metabolites. Using an abaxial sesamoid local anaesthetic model, it was determined that the highest no-effect dose (HNED) for its local anaesthetic effect was 2 mg. Using enzyme-linked immunosorbent assay (ELISA) screening, it was determined that subcutaneous (s.c.) administration of the HNED of mepivacaine to eight horses yielded a peak urinary concentration of apparent mepivacaine of 63 ng/mL 2 h after injection. The major identified metabolite recovered from equine urine after dosing with mepivacaine is 3-hydroxymepivacaine. Therefore, 3 hydroxymepivacaine was synthesized, purified and characterized, and a quantitative mass spectrometric method was developed for this metabolite as isolated from horse urine. Following subcutaneous injection of the HNED of mepivacaine, the concentration of 3-hydroxymepivacaine recovered from horse urine reached a peak of about 64.6 ng/mL at 4 h after administration as measured by GC/MS. The concentration of mepivacaine or its metabolites after administration of a HNED dose are detectable by mass spectral techniques. Within the limits of this research, the study suggests that recovery of concentrations less than about 65 ng/mL of 3-hydroxymepivacaine from post-race urine samples may not be associated with a recent LA effect of mepivacaine. PMID- 10372596 TI - Single-dose pharmacokinetics of flumequine in halibut (Hippoglossus hippoglossus) and turbot (Scophthalmus maximus). AB - Flumequine was administered to halibut (Hippoglossus hippoglossus) and turbot (Scophthalmus maximus) intravenously (i.v.) and orally (p.o.) at a dose of 10 mg/ kg bodyweight, and as a bath-treatment at a dose of 10 mg/L water for 2 h, using identical experimental designs. The study was performed in seawater with a salinity of 3% and a temperature of 10.3+/-0.4 degrees C (halibut) and 18.0+/-0.3 degrees C (turbot). Pharmacokinetic modelling of the data showed that flumequine had quite similar pharmacokinetic properties in halibut and turbot. Following intravenous administration, the volumes of distribution at steady state (Vss) were 2.99 L/kg (halibut) and 3.75 L/kg (turbot). Plasma clearances (Cl) were 0.12 L/kg (halibut) and 0.17 L/h x kg (turbot) and the elimination half-lives (t(1/2lambdaz)) were calculated to be 32 h (halibut) and 34 h (turbot). Mean residence times (MRT) were 25.1 h (halibut) and 22.2 h (turbot). Following oral administration, the t(1/2lambdaz) were 43 h (halibut) and 42 h (turbot). Maximal plasma concentrations (tmax) were 1.4 mg/L (halibut) and 1.9 mg/L (turbot), and were observed 7 h post administration in both species. The oral bioavailabilities (F) were calculated to 56% (halibut) and 59% (turbot). Following bath administration maximal plasma concentrations were 0.08 mg/L (halibut) and 0.14 mg/ L (turbot), and were observed 0 h (halibut) and 3 h (turbot) after the end of the bath. The bioavailability in halibut following a 2-h bath treatment was 5%. PMID- 10372597 TI - Pharmacokinetics and metabolism of ketoprofen after intravenous and intramuscular administration in camels. AB - The pharmacokinetics of ketoprofen were determined after an intravenous (i.v.) and intramuscular (i.m.) dose of 2.0 mg/kg body weight in five camels (Camelus dromedarius) using gas chromatography/mass spectrometry (GC/MS). The data obtained (median and range) following i.v. administration was as follows: the elimination half-life (t(1/2beta)) was 4.16 (2.65-4.29) h, the steady state volume of distribution (Vss) was 130.2 (103.4-165.3) mL/kg, volume of distribution (area method) (Vd(area)) was 321.5 (211.4-371.0) mL/kg, total body clearance (Cl) was 1.00 (0.88-1.08) mL/min x kg and renal clearance was 0.01 (0.003-0.033) mL/min x kg. Following i.m. administration, the drug was rapidly absorbed with peak serum concentration of 12.2 (4.80-14.4) microg/mL at 1.50 (1.00-2.00) h. The systemic availability of ketoprofen was complete. The apparent half-life was 3.28 (2.56-4.14) h. A hydroxylated metabolite of ketoprofen was identified by (GC/MS) under electron impact (EI) and chemical ionization (CI) scan modes. The detection times for ketoprofen and hydroxy ketoprofen in urine after an intravenous (i.v.) dose of 3.0 mg/kg body weight was 24.00 and 70.00 h, respectively. Serum protein binding of ketoprofen at 20 microg/mL was extensive; (99.1+/-0.15%). PMID- 10372598 TI - Cimetidine inhibits nitric oxide associated nitrate production in a soft-tissue inflammation model in the horse. AB - Cimetidine (CIM) is an H2-receptor antagonist that has been used in racehorses in an attempt to reduce the occurrence of stress-related gastric ulceration. It has also been shown to produce several useful effects other than its gastric acid suppression properties. Further, it is a well documented antagonist of cytochrome P-450 (CYP) mediated oxygenation reactions. Nitric oxide (NO), a recently discovered mediator or modifier of numerous physiological functions, is generated by several forms of nitric oxide synthase (NOS), one of which is inducible (iNOS). Inducible NOS, expressed in neutrophils and macrophages as part of the inflammatory response to noxious stimuli, contains both a CYP and a CYP reductase domain. Because of the similarity of structure of iNOS and CYP, it was decided to determine whether CIM could reduce NO production, using a carrageenan inflammation model in the horse. Two experiments were conducted. In Trial 1, six female Thoroughbred horses each had three tissue chambers inserted subcutaneously on the sides of the neck. The study was divided into three treatments: 0.9% NaCl (NaCI), CIM (3 mg/kg), and aminoguanidine (AG; 25 mg/kg), an inhibitor of iNOS. Each mare received three i.v. injections 12 h apart prior to instillation of 1 mL of carrageenan into the test chamber. Blood and tissue chamber fluid (TCF) were collected serially. Concentrations of NO3- (the major metabolite of NO), albumin, total protein, CIM and AG were measured and complete cell counts and differentials were conducted. Trial 2 also used six female Thoroughbred horses implanted with at least two tissue chambers inserted subcutaneously on the sides of the neck. The study was divided into two treatments: NaCl (0.9%) and CIM (6 mg/kg). Each mare received seven i.v. injections of either NaCl or CIM 8 h apart prior to instillation of 1 mL of carrageenan into the test chamber. Blood and TCF were collected serially as before, and analysed for NO3- and CIM content. Areas under the curve (AUC) of the different parameters were calculated for the periods of -1-1, -1-3 and -1-7 days (Trial 1) and -2-1 for Trial 2. Absolute values were also compared at 4, 8 and 12 h postcarrageenan. Saline treatment did not reduce the elevated concentrations of NO3- in either plasma or TCF. Plasma, test chamber and control chamber NO3-concentrations rose from 0 to 12 h, and were very similar in all three sampled fluids. Cimetidine significantly (P< or =0.05) decreased NO3 production in plasma over the periods of -1-1, -1-3, and -1-7 days post inflammation when compared to NaCl treatment in Trial 1. Aminoguanidine and CIM decreased NO3-production in TCF for the periods -1-1, 1-3, and -1-7 days post inflammation in Trial 1 and -2-1 for Trial 2. Both CIM and AG also significantly reduced NO3-concentrations in plasma and TCF at 12 h postinitiation (Trials 1 and 2). Thus CIM, at the doses studied, was capable of reducing NO3- concentrations in this model as effectively as AG, a relatively specific inhibitor of iNOS activity. PMID- 10372599 TI - Biliary elimination kinetics and tissue concentrations of oxytetracycline after intravenous administration in hens. AB - The pharmacokinetics of the biliary elimination of oxytetracycline (OTC) and tissue concentrations in certain organs were studied in 10 Leghorn hens. The animals were anaesthetized using xylazine/ketamine administered by the intramuscular (i.m.) route and were immobilized for right laparotomy. Both bile ducts were cannulated and a dose of 20 mg/kg of oxytetracycline hydrochloride was administered intravenously (i.v.). Samples of bile excreted were taken at predetermined intervals during 6 h. At 6 h animals were slaughtered and tissue samples of blood, liver, kidney, pancreas, spleen, heart, lung and pectoral muscle were taken. The values for OTC biliary elimination rate times were best fitted to a one-exponential equation. The maximum value for OTC biliary excretion rate (3.69+/-0.6 microg/min/kg) was reached at approximately 17.5 min (time to maximum concentration (tmax)). The first-order rate constant for the biliary excretion (k) and the half-life (t1/2) were 6.7x10(-3) min(-1) and 110.55 min, respectively. The mean value of area under the biliary excretion rate time curve (AUC) indicated that 839.77 microg/kg body weight (b.w.) were eliminated by the biliary route. The cumulative biliary excretion data indicated that approximately 4.20% of the dose was eliminated by this route, 3.28% being eliminated during the first 6 h and 0.92% thereafter. The highest mean concentrations were found in the kidney (35.82 microg/kg) and liver (16.77 microg/g). Significant differences were found between the concentrations of the various tissues studied. Plasma concentration was lower than that of the other tissues (except lung). PMID- 10372600 TI - The evaluation of the role of CCK in the opioid modulation of the motility of the gastrointestinal tract in sheep. AB - The participation of central cholecystokinin-8 (CCK-8) receptors in the modulatory effect of D-Ala2, N-Me-Phe4, Gly5-ol enkephalin (DAGO), a selective mu opioid receptor agonist, on the spike burst activity of the gastrointestinal tract (rumen, reticulum, antrum, duodenum, colon and caecum) in sheep was investigated. DAGO was infused intracerebroventricularly (i.c.v.) at doses of 0.1 1 microg/kg body weight (BW). It was shown that DAGO significantly inhibited myoelectrical activity of the wall of the forestomachs, abomasum and colon but stimulated this activity in the duodenum (rate of myoelectrical migrant complex MMC). The effects of DAGO were prevented by CCK-8 antagonists (L-364.718 and L 365.260) previously infused at doses of 5-20 microg/kg BW. The results of this present study indicate that central receptors of CCK-8 participated in the modulatory action of an opioid on myoelectrical activity of the gastrointestinal tract in sheep. Furthermore, this result suggests that CCK-8 is released in response to mu-receptor stimulation, because CCK-8 antagonists (L-364.718 and L 365.260) prevented the modulatory action of DAGO on the gastrointestinal motility in sheep. PMID- 10372601 TI - Treatment-resistant schizophrenia and beyond: current concepts and future prospects. Proceedings of a meeting. London, United Kingdom, July 8-9, 1998. PMID- 10372602 TI - The evolving definition of treatment resistance. AB - Despite the introduction of antipsychotic treatment for schizophrenia, the outcome for many patients has remained poor. This is largely due to the treatment resistant nature of schizophrenia in some patients and inadequate long-term maintenance treatment. The definition of treatment resistance remains controversial in spite of its importance. This review discusses the importance of treatment resistance and the factors affecting its definition in the light of recent advances in knowledge and treatment. A decade ago, positive symptoms were thought to be the prime outcome measure for schizophrenia and were the standard by which treatment resistance was largely assessed. More recently, however, a wider range of outcome measures has been recognized, including both negative symptoms and cognitive function. All of these outcome measures affect quality of life such that the patient may consider any outcome other than a return to premorbid levels of functioning as inadequate. Furthermore, patient responsiveness should be recognized as a continuum rather than as a dichotomy of response or nonresponse; partial response to treatment may not be accepted as satisfactory. Definitions of treatment resistance should reflect these factors. Patients may benefit from pharmacotherapy with atypical antipsychotics even if they do not meet criteria for narrowly defined treatment resistance. Although clozapine use has often been restricted to treatment-resistant patients, the benefit it bestows outweighs the potential risk of side effects in patients with less stringently defined treatment resistance. PMID- 10372603 TI - Pathophysiologic mechanisms in the pathogenesis and clinical course of schizophrenia. AB - It is widely accepted that schizophrenia originates from abnormalities occurring during the early stages of neural development. Although large studies have revealed behavioral precursors of schizophrenia in childhood, the disorder is usually not evident until patients are in their 20s or 30s. Some patients will be resistant to typical antipsychotic treatment at this first-onset of schizophrenia; however, treatment resistance develops in the majority of patients during the course of successive episodes. This ongoing deterioration suggests that a degenerative process operates during the active psychotic phase of the illness. This review presents evidence of neurodevelopmental and neurodegenerative mechanisms for the development of schizophrenia. These data indicate the importance of effective treatment at the first onset of schizophrenia to improve patient outcome. In addition, animal studies suggest that treatment with clozapine may prevent the neurodegenerative component responsible for the development of treatment resistance. PMID- 10372604 TI - Management strategies for the treatment of schizophrenia. AB - Considerable progress has been made during the past 10 years in the treatment of schizophrenia; however, the disease still represents a great challenge for the clinician. Frequently encountered problems include the patient who is only partially responsive to treatment or is treatment resistant and long-term relapse prevention. Patient compliance, long-term efficacy, drug dose, safety, and the duration of treatment are all important factors determining the degree of success of maintenance treatment in the prevention of relapse. This review discusses those aspects that should affect the clinicians' choice of treatment, including the recent introduction of atypical antipsychotics such as clozapine. PMID- 10372605 TI - Some adverse effects of antipsychotics: prevention and treatment. AB - Antipsychotic medication causes a wide range of adverse effects, which can be serious and may further imperil both the physical and psychological health of schizophrenic patients. The range of side effects patients commonly encounter includes weight gain, endocrine disturbances, sedation, anticholinergic effects, hypotension, seizures, and extrapyramidal symptoms. Less common and unpredictable reactions are blood dyscrasias, cardiotoxicity, sudden death, and the neuroleptic malignant syndrome. Antipsychotic drugs differ significantly regarding their propensity to cause these reactions. Patients should undergo comprehensive health checks before an antipsychotic is prescribed, and drug therapy should be individualized to take account of any preexisting symptoms. Side effects and the wider implications of drug treatment, such as effects on occupational and social functioning, should be discussed with the patient before initiating therapy. Patients should be regularly monitored for side effects during treatment and switched to alternative therapy if side effects are serious and/or persistent. PMID- 10372606 TI - A historical perspective of clozapine. AB - Having challenged the established view that extrapyramidal symptoms are an intrinsic feature of antipsychotic activity, clozapine was developed as the first atypical antipsychotic with activity against both the positive and negative symptoms of schizophrenia. Following its partial withdrawal due to concerns over agranulocytosis, clozapine was reintroduced in response to pressure from psychiatrists and is now used worldwide in patients with treatment-resistant schizophrenia, having demonstrated its superiority over typical antipsychotic agents. This, combined with its low propensity to cause tardive dyskinesia, has transformed the management of patients with schizophrenia. This article outlines the history of clozapine's development, from its discovery in 1958 to its current position as the "gold standard" therapy for treatment-resistant schizophrenia. PMID- 10372607 TI - The effects of clozapine on cognitive functioning in schizophrenia. AB - Cognitive function may be markedly impaired in patients with schizophrenia; however, it has only recently been recognized as an important factor in determining patient outcome. Research has shown that improvements in cognitive functioning occur independently of improvements in positive or negative clinical symptoms, and whereas typical antipsychotics may improve clinical symptoms, they have little or no efficacy in improving cognitive dysfunction. However, there is evidence that the atypical antipsychotic clozapine may improve this core deficit of schizophrenia. This review summarizes 12 published studies that assessed the effect of clozapine on cognitive functioning. As a group, these studies suggest that psychomotor speed, verbal fluency, and verbal learning and memory may be improved by treatment with clozapine. Such cognitive improvements with clozapine treatment may offer an advantage to patients with schizophrenia by enhancing the possibility of better vocational functioning and quality of life. PMID- 10372608 TI - Clozapine: a comparison with other novel antipsychotics. AB - Clinical studies with clozapine have clearly demonstrated its superior efficacy over that of conventional antipsychotics in treatment-resistant schizophrenic patients. In comparative trials with these drugs, considerably more patients respond to treatment with clozapine than to conventional antipsychotic medication. Recently, new antipsychotics, such as olanzapine, quetiapine, risperidone, sertindole, and zotepine, have been introduced, but extensive data on their effects in treatment-resistant patients are not yet available. Published studies have drawn criticism in terms of inappropriate titration schedules, nonequivalent dosing between treatment groups, short treatment duration, and inadequate sample sizes. Further research will be needed to determine whether novel antipsychotics may substitute for clozapine in the future or whether clozapine will retain its unique role in the management of patients suffering from difficult-to-treat schizophrenic disorders. PMID- 10372609 TI - Optimizing clozapine treatment. AB - Compliance with conventional antipsychotic medication is often poor, with many patients discontinuing treatment only a few months after commencing therapy. The side effects of treatment, which are not necessarily restricted solely to motor symptoms, are often considered to be responsible for this noncompliance. In contrast to conventional antipsychotics, clozapine is associated with only minimal extrapyramidal symptoms, and in most patients, its use results in significant improvements in compliance. However, clozapine does induce a variety of adverse effects, most of which are of limited duration and either preventable or manageable if a number of simple clinical procedures are followed. Clozapine therapy is associated with a beneficial risk/benefit ratio in the majority of treatment-resistant schizophrenic patients. With careful hematologic control, the risk of agranulocytosis can be minimized. The marked increase in the well-being of patients receiving clozapine should stimulate psychiatrists to broaden its use and not limit it to severely treatment-resistant individuals. PMID- 10372610 TI - Clozapine: the commitment to patient safety. AB - Clozapine represents the "gold standard" therapy for treatment-resistant schizophrenia including use for symptom reduction and use in patients intolerant of extrapyramidal side effects associated with other antipsychotics. Despite its clear benefit in these areas, its use has been associated with a serious, and sometimes life-threatening, risk for agranulocytosis. Effective white blood cell monitoring systems have been developed by Novartis affiliates across the world to ensure its safe use and to meet local health standards. The goals of the monitoring programs include: (1) weekly white blood cell monitoring during the initial months of therapy for early detection of severe leukopenia; (2) immediate discontinuation of clozapine if severe leukopenia is observed; (3) exclusion from reexposure to clozapine if a patient experiences clozapine-induced agranulocytosis; and (4) early cessation of treatment if hematologic guidelines are not followed ("no blood, no drug" policy). Together, these systems have demonstrated a worldwide reduction in the observed rate of agranulocytosis and in fatalities related to the emergence of agranulocytosis when rigorous monitoring systems are in place. PMID- 10372611 TI - The effects of clozapine on aggression and substance abuse in schizophrenic patients. AB - Aggressive behavior in schizophrenic patients, although infrequent, is a serious problem. It is, however, a relatively common reason for psychiatric admission and poses an increasing threat as more patients are cared for in the community. There is a strong association between substance abuse and violent behavior, and comorbid substance abuse in schizophrenia is also a major problem. The recent introduction of the atypical antipsychotics has brought hope for the pharmacologic management of this group of patients. These newer agents are thought to have antiaggressive effects and perhaps decrease cravings for illicit substances and alcohol. Data from a number of studies have demonstrated that clozapine has antiaggressive effects. A retrospective analysis of 331 schizophrenic patients assessed the effects of clozapine on hostility and aggression. At baseline, 31.4% of patients showed overt physical aggression, and after an average of 47 weeks of treatment with clozapine, this rate had fallen to 1.1%. The antiaggressive effects of clozapine were relatively specific and could not be explained by sedation or general antipsychotic effects. These effects were more pronounced than the effects on other symptoms and were also present in those patients who showed the highest pretreatment levels of hostility and aggression. Clozapine may also be of benefit in the treatment of schizophrenic patients with comorbid substance abuse. After 6 months of treatment with clozapine, substance abusers and nonabusers with schizophrenia or schizoaffective disorder showed similar improvements on measures of psychopathology and psychosocial functioning. PMID- 10372612 TI - Suicide and schizophrenia: clozapine and the InterSePT study. International Clozaril/Leponex Suicide Prevention Trial. AB - Suicide is one of the most serious of schizophrenic symptoms and claims the life of 9% to 13% of patients. The annual rate of suicide in schizophrenic patients is reported to be in the range of 0.4% to 0.8%, a rate that has remained constant despite the introduction of antipsychotic therapy and attendant case-management systems. The risk of suicide is not significantly different in neuroleptic resistant or -responsive schizophrenic patients. A study of 421 schizophrenic patients reported no significant difference in the incidence of lifetime and current episodes of suicidality in treatment-resistant and -responsive patients. A number of studies with clozapine, an atypical antipsychotic, have demonstrated an 80% to 85% reduction in suicide in neuroleptic-resistant patients. This is accompanied by a decrease in depression and psychopathology and improved cognition. Clozapine's modulation of serotonergic, noradrenergic, cholinergic, and dopamine function may be the biological basis for the reduction in suicide. Weekly contact with patients, for white blood cell monitoring, has also been put forward as one explanation. To further confirm suicide risk reduction as a benefit of clozapine therapy, the International Clozaril/Leponex Suicide Prevention Trial (InterSePT) is currently being conducted. This large, prospective treatment study will compare the rate of suicide attempts and completions in schizophrenic patients at high risk of suicide randomly assigned to receive clozapine or olanzapine. The bias of weekly visits will be excluded. Results should be available in 2001. PMID- 10372613 TI - Treatment of occult pneumothoraces from blunt trauma. AB - BACKGROUND: Occult pneumothoraces (OPTXs) are seen on abdominal computed tomographic (CT) scans but not on routine chest x-ray films. Optimal treatment for blunt trauma OPTXs has not been defined. We hypothesized that OPTXs could be safely observed without need for a chest tube (CT). METHODS: A prospective trial randomized blunt trauma patients with OPTXs to CT scan or observation. Patients were not excluded for positive pressure ventilation. Primary outcome measures were respiratory distress and pneumothoraces progression. RESULTS: Thirty-nine patients with 44 pneumothoraces were enrolled. Eighteen patients received a CT scan, and 21 patients were observed. Nine patients in each group received positive pressure ventilation. There was no difference in overall complication rate. No patient had respiratory distress related to the OPTX or required emergent CT scan. CONCLUSIONS: Observation of OPTX is not associated with an increased incidence of pneumothorax progression or respiratory distress. These pneumothoraces can be safely observed in patients with blunt trauma injury regardless of the need for positive pressure ventilation. PMID- 10372614 TI - Duodenal versus gastric feeding in ventilated blunt trauma patients: a randomized controlled trial. AB - OBJECTIVE: To evaluate transpyloric feeds as they have been proposed as a means of providing enteric nutrition more rapidly and minimizing morbidity in ventilated trauma patients. METHODS: Between July of 1994 and June of 1997, 80 adult ventilated trauma patients were enrolled in a randomized controlled trial of duodenal versus gastric feeds. Feeding was initiated within 72 hours of injury. RESULTS: Forty-three patients received gastric feeds (G), and 37 patients received duodenal feeds (D). Mean age was 34.7+/-15.7 years (G) and 33.6+/-17.5 years (D); the difference in age was not significant (NS). Mean Injury Severity Score was 30.0+/-11 (G), 33.0+/-9.7 (D), NS. Mean Acute Physiology and Chronic Health Evaluation (APACHE II) score was 18.0+/-6.0 (G) and 18.0+/-7.4 (D), NS. Thirty-four of 43 patients were men (G) and 28 of 37 patients were men (D), NS. Use of narcotics and paralytics between the two groups was not significantly different. Energy requirements were 1.4 times basal energy expenditure at 2,127+/ 304 Kcal (G) and 2,089+/-274 Kcal (D), NS. Intensive care unit length of stay was a median of 7 days (range, 3-32 days) (G) and 10 days (range, 3-24 days) (D), NS. Number of days on ventilator was a median of 5 days (range, 3-15 days) (G) and 9 days (range, 2-13 days) (D), NS. Hospital length of stay was a median of 25 days (range, 9-88 days) (G) and 30 days (range, 16-47 days) (D), NS. Recorded morbidity was not significantly different. Pneumonia rates were 42% (G) and 27% (D), NS. Time to tolerate full-strength feeds for 24 consecutive hours was 43.8 hours +/-22.6 (G) and 34.3 hours +/-7.1 (D), difference significant at p = 0.02. CONCLUSION: Length of stay and ventilator days were not significantly different. A larger trial would be required to determine differences in the rates of pneumonia <20%. Transpyloric-duodenal feeds significantly reduce the time required to achieve targeted enteric nutrition. PMID- 10372615 TI - Granulocyte colony-stimulating factor ameliorates life-threatening infections after combined therapy with barbiturates and mild hypothermia in patients with severe head injuries. AB - OBJECTIVE: The objective of this study was to clarify the effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) administration on infections in patients with severe head injuries after combined therapy with high dose barbiturates and mild hypothermia. PATIENTS AND METHODS: Since 1996, we have administered rhG-CSF to eight patients with severe head injuries for 5 days (group G). Their treatment results were compared with those of 22 patients cared for earlier without rhG-CSF treatment (group N). All patients in both groups met the criteria of total leukocyte count (TLC) less than 5,000/mm3, C-reactive protein (CRP) over 10 mg/dL, and the presence of an infectious complication. Changes in the TLC, CRP, respiratory index, intracranial pressure, and infectious condition were evaluated in both groups. In addition, the nucleated cell count and differentiation from bone marrow aspiration, neutrophil functions, serum concentrations of interleukin-6, and plasma concentration of leukocyte elastase were evaluated in group G. RESULTS: In group G, TLC, nucleated cell count, and neutrophil functions significantly increased, whereas CRP, respiratory index, and interleukin-6 decreased reciprocally. There was no deterioration of intracranial pressure and leukocyte elastase. Consequently, seven of the eight patients in group G recovered from life-threatening infections, and none of the eight patients died. However, in group N, CRP and respiratory index remained high and TLC did not increase as much as it did in group G. Infections continued after 5 days in 17 of the 22 patients, 7 of whom died from severe infections during hospitalization. CONCLUSION: Administration of rhG-CSF ameliorated life threatening infections without causing lung injury or increasing brain swelling in patients with severe head injuries who were treated with combined therapy involving high-dose barbiturates and mild hypothermia. PMID- 10372616 TI - Physiologic effects of externally applied continuous negative abdominal pressure for intra-abdominal hypertension. AB - BACKGROUND: To determine the ability of an externally applied continuous negative abdominal pressure device (CNAP) to reverse the effects of elevated intra abdominal pressure on the central nervous and cardiovascular systems. METHODS: Anesthetized, ventilated swine had catheters placed for measurement of intra abdominal (IAP), intracranial (ICP), central venous, pulmonary artery, pulmonary artery occlusion, mean arterial, peak inspiratory, inferior vena cava, and femoral vein pressures. After the animals stabilized, baseline measurements were obtained. IAP was increased by incrementally instilling an isosmotic polyethylene glycol solution into the peritoneal cavity until it was 25 mm Hg above baseline. IAP was maintained at 25 mm Hg above baseline for 2 hours. CNAP was then applied for 2 hours. All parameters were remeasured 30 minutes after each increase in IAP, at 2 hours after attaining maximum IAP, and lastly at 2 hours after abdominal decompression. Cardiac index was maintained near baseline by volume expansion. RESULTS: Elevation of IAP to 25 mm Hg above baseline for 2 hours caused increases (p<0.05) in central venous pressure (10.3+/-0.9 to 15.2+/-1.7), inferior vena cava pressure (13.0+/-1.0 to 29.5+/-1.5), femoral vein pressure (13.5+/-0.5 to 33.3+/-1.3), ICP (10.6+/-1.5 to 21.0+/-1.5), and peak inspiratory pressure (18.3+/-0.3 to 34.2+/-1.0). The mean arterial pressure (106.3+/-3.5 to 125.8+/-3.4), pulmonary artery pressure (24.3+/-2.3 to 31.3+/-1.7), and pulmonary artery occlusion pressure rose (12.3+/-0.9 to 17.5+/-3.5), but not significantly. Cardiac index (3.3+/-0.5 to 3.4+/-0.4) remained essentially unchanged. CNAP significantly (p<0.05) decreased IAP (30.7+/-1.3 to 18.2+/-1.3), central venous pressure (15.2+/-1.7 to 12.4+/-2.1), inferior vena cava (29.5+/-1.5 to 19.2+/ 1.3), and ICP (21.0+/-1.5 to 16.2+/-1.3). Pulmonary artery occlusion pressure (17.5+/-3.5 to 15.0+/-3.1) and peak inspiratory pressure (34.2+/-1.0 to 29.7+/ 1.1) decreased, but not significantly. CONCLUSION: Acutely elevated IAP causes a significant increase in ICP and impaired cardiovascular and pulmonary function. Abdominal decompression remains the standard of care for abdominal compartment syndrome. However, in patients in whom an increased IAP does not require surgical decompression, the results of this study suggest that externally applied CNAP may be of value. PMID- 10372617 TI - Prospective evaluation of the potential role of teleradiology in acute interhospital trauma referrals. AB - BACKGROUND: Teleradiology is one form of telemedicine that would allow the transmission of radiographs before the transfer of acutely traumatized patients between referring and receiving hospitals. The purpose of this study was to evaluate the potential impact of a prehospital teleradiology system on trauma patient management and transfer. METHODS: Forty-four injured adults referred to a trauma center were included. The history, physical examination, and radiographic findings reported by the referring physician to the receiving physician were documented. The plain radiographs of the chest, pelvis, and cervical spine taken at the referring hospital were obtained after patient transfer. For each case, two reviewers blinded to the case (surgeon [S] and emergency department physician [E]) and one reviewer not blinded to the case were individually presented with the referring physician's report and the radiographs. The reviewers were surveyed as to the implications of viewing the plain radiographs taken at the referring hospital before patient transfer. RESULTS: Overall, the blinded reviewers felt that viewing the radiographs before transfer would have influenced care in 40% and 38% of cases as judged by (S) and (E), respectively, with a crude agreement of 67.5% (kappa level, 0.32). The blinded reviewers (S and E) commonly noted the following four changes in management as a result of viewing the referred radiographs: requested further clinical history (S, 18%; E, 23%), suggested further pretransfer interventions (S, 38%; E, 30%), suggested further pretransfer diagnostic tests (S, 25%; E, 13%), and emphasized precautions during transfer (S, 28%; E, 30%). The nonblinded reviewer suggested potential influence in the management of at least 65% of the cases. CONCLUSION: This study suggests that viewing the radiographs of acutely injured trauma patients has the potential to influence many aspects of the management of interhospital transfer. PMID- 10372618 TI - Vertical shear injuries: is there a relationship between residual displacement and functional outcome? AB - BACKGROUND: Residual vertical displacement is often cited as being related to poor outcome in patients with pelvic injuries. This study attempts to clarify the relationship between residual vertical displacement and functional outcome. METHODS: From 1982 to 1989, over 500 patients with pelvic ring injuries were treated at two Level I trauma centers. Thirty-three patients with vertical shear (Tile C) fractures and residual displacement (2-52 mm) were evaluated. Outcomes were quantified by using SF-36 Short-Form Health Survey (SF-36) and the Iowa Pelvic Score (IPS). RESULTS: There was no correlation between IPS or SF-36 scales and residual vertical displacement. The IPS correlated (p<0.05) with seven of eight SF-36 categories, excluding mental health. Patients reporting limp and leg length discrepancy also correlated with the IPS and select SF-36. CONCLUSION: Pelvic injuries showed no correlation between functional outcome and residual vertical displacement suggesting other factors. The degree of residual vertical displacement does not affect functional outcome. PMID- 10372619 TI - Free flap reconstructions of 100 tibial fractures. AB - OBJECTIVE: Reconstructive microsurgery has been part of the treatment for severe tibial fractures for over 20 years. METHODS: In this study, we have analyzed 100 patients with 104 microvascular reconstructions to the lower leg in a 15-year period (1980-1994). Sixty-three of the reconstructions were made during the past 5 years (1990-1994), which means that microvascular reconstruction is increasing as a treatment of severe tibial fractures. RESULTS: Free flap transfer is a safe procedure: the failure rate among the patients studied was 2%, and the amputation rate was 5%. CONCLUSION: In the past 5 years, the flap survival rate and the microvascular free flap operation methods were the same as they were in the 1980s, but the methods for enhancing the fracture union or reconstructing the bone defect has changed. PMID- 10372620 TI - Fixation of small tibial avulsion fracture of the posterior cruciate ligament using the double bundles pull-through suture method. AB - BACKGROUND: Avulsion of the tibial insertion of the posterior cruciate ligament is commonly repaired via open reduction and internal fixation with a screw, Kirschner's wire, and suture. In the case of a major bony fragment, this technique is adequate to achieve rigid fixation. In the case of an avulsion fracture with a small bony fragment, however, it is not uncommon to break the bone fragment during screw fixation. We describe a new technique for fixation of an avulsion fracture with a small bony fragment. The technique uses a double bundles pull-through suture technique that repairs the anterolateral and posteromedial components of the posterior cruciate ligament simultaneously. METHODS: From March 1994 through May 1997, 12 patients with small tibial avulsion fractures of the posterior cruciate ligament were treated using this technique. RESULTS: At an average of 18 months after surgery (range, 12-24 months), the preliminary clinical and radiographic results were satisfactory. Eleven patients could return to the same or a higher level of preinjury sports activity. According to the International Knee Documentation Committee rating system, 10 of the 12 patients had normal or nearly normal ratings. CONCLUSION: The double bundles pull-through suture technique can avoid the risk of breakage of the small bony fragment, does not require the removal of hardware, and can achieve adequate repair in the anatomic situation. Our clinical experience suggests that it is a good choice for fixation in cases of avulsion fracture with a small bony fragment. PMID- 10372621 TI - One or two lag screws for fixation of Danis-Weber type B fractures of the ankle. AB - BACKGROUND: There are many limitations in using the plate for fixation of lateral malleolar fractures of the ankle. METHODS: This study prospectively reviewed 60 of 72 consecutive patients with an average follow up of 3.8 years who had Danis Weber type B ankle fractures that were treated with one or two 3.5-mm cortical lag screws without plate fixation. RESULTS: A stable anatomic reduction was obtained in 57 cases (95%). The average time to healing by radiograph was 3.1 months. There were two delayed unions and no nonunions. Joint space narrowing was found in one case. Clinical assessment was satisfactory in 56 of the patients (93%). CONCLUSION: Noncomminuted Danis-Weber Type B ankle fractures can be successfully treated without the risk of lateral plating, which includes cartilage damage caused by penetration of the ankle joint, increased periosteal dissection, and the risk of subsequent hardware removal. PMID- 10372623 TI - Aggressive treatment of 119 open fracture wounds. AB - BACKGROUND: The purpose of this study was to determine whether immediate primary closure of open fracture wounds can be performed without increasing the incidence of infections and delayed unions/nonunions. Although the traditional management of these injuries has been open treatment, a trend toward immediate primary closure has evolved on our service. METHODS: All open fractures presenting to an urban Level I trauma center during a 42-month period were reviewed. Of the 127 patients with open fractures, 90 patients (119 open fractures) were initially treated at the above institution within 24 hours of injury, had fractures proximal to the carpus or tarsals, and were followed-up until fracture union. All patients underwent emergent wound irrigation and debridement. The method of fracture immobilization and timing of wound closure was left to the discretion of the attending orthopedic surgeon. Immediate primary closure was used in 22 of 25 Grade I open fractures (88%), 37 of 43 Grade II fractures (86%), 24 of 32 Grade IIIa fractures (75%), 4 of 12 Grade IIIb fractures (33%), and 0 of 7 Grade IIIc fractures (0%). RESULTS: Eight fractures (7%) were complicated by a deep wound infection/osteomyelitis, and 19 fractures (16%) developed a delayed union/nonunion. Statistical analysis revealed no significant difference in delayed/nonunion and infection rates between immediate and delayed closures. CONCLUSION: Immediate primary closure of open fracture wounds after a thorough debridement by an experienced fracture surgeon appears to cause no significant increase in infections or delayed union/nonunions. In addition, early closure may decrease the requirement for subsequent debridements and soft-tissue procedures, thereby minimizing surgical morbidity, shortening hospital stays, and reducing costs. We feel that a randomized, prospective study of this aggressive approach to open fracture care is warranted. PMID- 10372622 TI - Diaphyseal forearm fractures treated with and without bone graft. AB - BACKGROUND: The purpose of this study was to determine whether the acute bone grafting of diaphyseal forearm fractures decreases the incidence of nonunion and reduces the time to union. Although the traditional treatment of comminuted radius and/or ulnar shaft fractures involves bone graft, a recent report called into question this practice. PATIENTS: A database search was used to identify all acute diaphyseal forearm fractures presenting to an urban Level I trauma center between 1988 and 1996. All radius and/or ulnar shaft fractures, as well as all Monteggia and Galeazzi fracture-dislocations, in patients with closed physes were included. The charts and operative reports were available for 64 diaphyseal forearm fractures in 49 patients. Fifty-six fractures were followed for at least 1 year beyond clinical and radiographic union. The injuries were treated with open reduction and plate fixation by experienced orthopedic traumatologists. All noncomminuted fractures were treated without bone graft. For the comminuted fractures, the decision to use bone graft was left to the discretion of the operating surgeon. RESULTS: Overall, 55 of 56 fractures (98%) achieved union at a mean of 49 days (range, 19-123 days), with the only nonunion occurring in a patient with a closed, noncomminuted Galeazzi injury. Among the 20 noncomminuted fractures, all of which were treated without bone graft, 19 (95%) achieved union at a mean of 50 days (range, 19-102 days). Among the 36 comminuted fractures, all 25 treated without bone graft achieved fusion at an average of 50 days (range, 20 123 days) and all 11 treated with bone graft achieved union at an average of 45 days (range, 22-67 days). No statistically significant difference in the incidence of nonunion or time to union was noted between fractures that were treated with and without bone graft. CONCLUSION: Acute bone grafting of diaphyseal forearm fractures did not affect the union rate or the time to union. PMID- 10372624 TI - Can the distance fallen predict serious injury after a fall from a height? AB - BACKGROUND: After a fall, the distance fallen is sometimes used to predict the injury severity. We aimed to examine how distance fallen performs as a predictor of major injury. METHOD: A cohort of trauma victims attending our emergency department after having fallen from a height was identified retrospectively, and data were collected regarding the fall and injuries sustained. Performance of threshold heights, ranging from 2 meters (6.6 feet) to 10 meters (32.8 feet), as a diagnostic test for major injury was assessed. RESULTS: Height fallen performed poorly over the range of thresholds used. At low thresholds, sensitivity was inadequate to rule out major trauma, whereas the low prevalence meant that, despite impressive specificity at higher thresholds, positive predictive value was poor. At the optimal threshold of 5 meters (16.4 feet), the positive predictive value was 0.17 and sensitivity was 0.33. CONCLUSION: Height of fall is a poor predictor of major injury. PMID- 10372625 TI - Snowboarder's wrist: its severity compared with Alpine skiing. AB - BACKGROUND: Although the upper extremity, especially the wrist, has been reported to be the most commonly injured site in snowboarding, the severity of these injuries is still unknown. The purpose of this study is to compare the severity of wrist injuries in snowboarding with those in alpine skiing for insight into the treatment of snowboarder's wrist. MATERIALS AND METHODS: The cases of 11,598 patients injured while snowboarding and skiing who presented to the Zao clinic during the past 7 seasons were reviewed and compared. Demographics were studied, focusing on fractures around the wrist joint. Roentgenographically precise assessment of the distal radial fracture was performed according to AO classification. RESULTS: Snowboarders were more likely to injure the wrist than were skiers (18.7% vs. 2.5%, p<0.01). In these wrist injuries, distal radial fractures occurred at a rate of 0.28 per 1000 snowboarders and 0.008 per 1000 skiers. Comminuted and articular fractures classified as AO type A3, B and C, which required surgical treatment, were 49.4% of distal radial fractures in snowboarders and 23.8% in skiers. CONCLUSION: Wrist injury sustained while snowboarding is characterized as a severe and complex injury. Thus, we call attention to its severity in the treatment of snowboarder's wrist. PMID- 10372626 TI - Snowboarding injuries of the chest: comparison with skiing injuries. AB - BACKGROUND: Snowboarding injuries have become more common with the remarkable increase in the sport's popularity. However, although there are many reports of orthopedic injuries caused by snowboarding, there are few reports on injuries to the chest. In this study, we attempted to identify the characteristics of snowboarding injuries of the chest in comparison with alpine skiing injuries. METHODS: Between December of 1988 and April of 1997, 1,579 and 9,108 patients were treated for snowboarding and skiing injuries, respectively. All patients were initially examined by emergency physicians who used chest x-ray film. Patients with known or suspected chest injuries were further examined by using chest computed tomography and ultrasonography by thoracic and general surgeons. A total of 96 snowboarding patients and 247 skiing patients had chest injuries. RESULTS: The chest injuries among snowboarders accounted for 6.1% of all injuries compared with only 2.7% amongst skiers. Snowboarders with chest injuries were younger, more often beginners, and more frequently occurred during the afternoon than skiers. Several distinct patterns of injury were noted among these two groups. As the cause of injury, a riding mistake during jumping was significantly higher for snowboarders (50.0%) than for skiers (0%). The incidence of rib fracture during snowboarding (55.2%) was significantly higher than during skiing (41.3%). There were no mortalities in either group. CONCLUSION: A riding mistake during improper jumping may be the primary cause of chest snowboarding injuries. Furthermore, snowboarders are much more likely to injure the chest, particularly by rib fractures, than skiers. PMID- 10372627 TI - Snowboard head injury: prospective study in Chino, Nagano, for two seasons from 1995 to 1997. AB - BACKGROUND: The popularity of snowboarding has been growing rapidly throughout the world. To date, however, the risk of head injury associated with this relatively new winter sport, especially in comparison with alpine skiing, has not been well analyzed. This study was conducted to assess the risk of head injury in snowboarding and to elucidate its features in comparison with skiing head injury. METHODS: We prospectively analyzed 301 cases of head injuries related to snowboarding or skiing experienced from December of 1995 to May of 1997 at our institution, which is located close to the most popular skiing areas in Japan. Of those injuries, 143 cases were snowboard related and 158 cases were ski related. In addition to appropriate medical evaluation and medical care, detailed examination was performed on every patient to determine various factors, including sex, age, skill level, cause and mechanism of the accident, and the side of impact to the head. The data are statistically analyzed to elucidate unique features of snowboard head injury. RESULTS: During the study period, 2.2 million snowboarders and 4.2 million skiers visited the five skiing facilities that are covered by our hospital. Thus, the incidence of head injury was 6.5 per 100,000 visits for snowboarders and 3.8 per 100,000 visits for skiers. Beginning snowboarders more frequently sustained head injuries compared with beginning skiers (60 of 142 vs. 48 of 154, p = 0.022). Likewise, frequent causes of snowboarding head injuries were fall during jumping (43 of 139 vs. 2 of 147, p<0.0001), falling backward (67 of 127 vs. 49 of 144, p = 0.001), and occipital impact (67 of 126 vs. 49 of 147). More importantly, there were nine major head injury cases (6.3%) in snowboard head injuries in contrast to only two such cases found in skiing head injuries (1.3%). Of 11 major head injury cases, 10 were caused by occipital impact. CONCLUSION: These results indicate that snowboarders, particularly beginners, are at higher risk for head injury, frequently involving occipital impact, and could lead to more major head injuries. We propose that measures should be taken to protect the head, especially the occiput, in snowboarding. PMID- 10372628 TI - Coagulofibrinolytic changes after isolated head injury are not different from those in trauma patients without head injury. AB - BACKGROUND: To test the hypothesis that tissue factor release, thrombin activation, fibrin formation, and fibrinolysis after an isolated head injury are equal to those in patients without head injury, as well as to investigate the precise time course of the coagulation and fibrinolytic abnormalities after head injury, we performed prospective and retrospective studies. METHODS AND RESULTS: In the prospective study, 5 patients with isolated head injury and 11 trauma patients without head injury took part in this study. Tissue factor antigen concentration, prothrombin fragment F1+2, thrombin antithrombin complex, fibrinopeptide A, and fibrin degradation products (D-dimer) were measured on the day of admission, and days 1, 2, 3, and 4 after admission. The levels of all five hemostatic molecular markers were markedly elevated on the day of admission, and then gradually decreased to day 4. The levels and the time course of these hemostatic markers in patients with isolated head injury were not different from those in the control patients. The same incidence of disseminated intravascular coagulation between the two groups was also observed. In the retrospective study, the records of fibrinopeptide Bbeta15-42, plasmin antiplasmin complex, plasminogen activator inhibitor-1 antigen concentration (PAI-1 antigen), and PAI 1 activity in 76 trauma patients were reviewed. On the basis of the exclusion criteria, 9 patients with isolated head injury and 30 control patients were selected for the study group. Fibrinopeptide Bbeta15-42 and plasmin antiplasmin complex markedly elevated on the day of admission, then decreased on day 1, and tended to increase to day 5. Markedly elevated PAI-1 antigen and PAI-1 activity on the day of admission significantly decreased on day 1 and recovered to the normal values on day 5. The changes of these molecular markers in patients with isolated head injury were equal to those in the control patients. CONCLUSION: We systematically elucidated the time course of coagulation and fibrinolysis after isolated head injury. We further demonstrated that changes in coagulofibrinolytic and antifibrinolytic systems in patients with isolated head injury are not different from those in patients without head injury. PMID- 10372630 TI - New instrument that uses near-infrared spectroscopy for the monitoring of human muscle oxygenation. AB - BACKGROUND: Early detection of vascular impairments after free tissue transfers are essential to prevent flap failure. Near-infrared spectroscopy showed good promise to monitor flaps at deep levels successfully without being invasive. The purpose of this study was to test whether the INVOS 3100 cerebral oxymeter is capable of detecting circulatory impairments. METHODS: In 10 healthy adults, near infrared spectroscopy was used to measure regional saturation values during tourniquet ischemia and venous outflow restriction, in two test cycles. The probe, containing an infrared-light-emitting source and two infrared-light sensors, was placed below the elbow above the brachioradialis muscle. Statistical comparison of the data was performed using the Friedman test and the Wilcoxon Wilcox test. RESULTS: Venous and arterial occlusions were characterized by an instantaneous fall of the regional saturation. Arterial occlusion showed a mean decrease of the saturation values down to 28+/-9%, whereas venous occlusion showed a mean fall of saturation values down to 51+/-12%. These falls were significant after 3 minutes of occlusion compared with baseline values (74+/-6%). The differences between arterial and venous occlusions were statistically significant. CONCLUSION: This study, designed to test less-expensive equipment, was able to measure absolute values, and was not prone to interference caused by probe movement, providing information on the oxygenation profile accurately and noninvasively, and distinguishing between arterial and venous occlusion. PMID- 10372629 TI - Wound infections after minor limb lacerations: risk factors and the role of antimicrobial agents. AB - BACKGROUND: The requirement for antimicrobial agents in patients with minor limb lacerations was prospectively studied. METHODS: The development of wound infections in patients with minor limb lacerations who received amoxicillin plus clavulanate acid treatment (group A, 52 patients) was studied and compared with patients who did not (group B, 48 patients). RESULTS: Wound infection occurred in 6 (11.5%) and 10 (21%) patients in groups A and B, respectively (p>0.10). Statistically significant risk factors for the development of infection were diabetes mellitus (odds ratio [OR], 15.8; p<0.001), lower limb lacerations (OR, 33.5; p<0.001), lacerations caused by compressive forces (OR, 21.6; p = 0.007), laceration length from 5 to 8 cm (OR, 7.04; p = 0.001), ragged laceration edge (OR, 2.55; p = 0.049), and skin tension (OR, 2.00; p = 0.006). CONCLUSION: The use of antimicrobial agents in minor limb injuries was not associated with a significant reduction of infection rate. Routine antimicrobial treatment is discouraged. PMID- 10372631 TI - Burn wound assessment in porcine skin using indocyanine green fluorescence. AB - BACKGROUND: An accurate assessment of deep dermal burns within the first week after burn is still an unresolved clinical problem. Infrared-excited fluorescence of indocyanine green was examined as a method of early determination of burn depth. METHODS: Burns of varying depths were placed on the paraspinal region, flank, and abdomen of swine using a heated brass block. Fluorescence images of the burns were recorded 1, 24, 48, and 72 hours later. RESULTS: The ratio of fluorescence in 64 burn wounds relative to adjacent normal tissue identified wounds that healed and did not heal within 21 days with an accuracy of 100%, after accounting for the age of the burn. Higher fluorescence ratios were observed in newly placed burns relative to older burns having comparable depths. CONCLUSION: Deep partial-thickness burns were differentiated from deep dermal full-thickness burns in a porcine skin burn model independent of body location. Diagnosis was possible between 1 and 72 hours after injury. PMID- 10372632 TI - Selectin blockade worsened lipopolysaccharide-induced lung injury in a swine model. AB - BACKGROUND: Polymorphonuclear leukocytes have been reported to play an important role in various acute lung injuries. Neutrophil recruitment into tissues is a multistep process involving sequential engagement of adhesion molecules. The objective of this study was to determine the effect of selectin inactivation with Sulfo Lewis C (SO3-3betaGal1-3betaGlcNAc-O(CH2)8-COOMe) on the pulmonary response to lipopolysaccharide (LPS) infusion. METHODS: All animals (n = 11) were pretreated with an intramuscular injection of a priming dose of Escherichia coli LPS (10 microg/kg). Eighteen hours later, animals received an intravenous infusion of LPS (20 microg/kg) over 20 minutes. All animals were resuscitated with a lactated Ringer's solution. Group I (G1; n = 5) received no additional treatment. Group II (G2; n = 6) received a bolus injection of Sulfo Lewis C (10 mg/kg) 10 minutes before LPS insult followed by a continuous infusion (1 mg/kg per hour) for the rest of the study. Animals were observed for 5 hours from initiation of the LPS infusion and killed. Cardiopulmonary variables and blood gases were measured serially. The multiple inert gas elimination technique (MIGET) was used to evaluate the matching of air flow and blood flow in the lung 5 hours after LPS infusion. Histologic evaluation of the parenchymal injury was performed by using light microscopy. The number of polymorphonuclear leukocytes and red blood cells in the alveolar spaces per field at 400x magnification were counted in 10 randomly selected fields. RESULTS: Hypoxemia, indexed as Pao2/FIO2, was exacerbated by the administration of Sulfo Lewis C (G1:437+/-33 vs. G2: 241+/ 63 mm Hg at 5 hours, p<0.03). This finding is supported by the multiple inert gas elimination technique analysis, which demonstrated significantly greater blood flow to true shunt in G2 (G1:4.42+/-1.75 vs. G2:23.2+/-5.69, p<0.02). There was no difference between the two groups in red blood cell counts in the alveolar spaces. However, polymorphonuclear leukocyte counts were significantly greater in G2 (G1:1.8+/-0.58 vs. G2:9.9+/-2.34, p<0.01). CONCLUSION: Selectin blockade significantly worsened lung injury induced by LPS infusion, and greater numbers of neutrophils were observed in alveolar spaces in the group treated with Sulfo Lewis C. These findings are supported by the multiple inert gas elimination technique analysis, which demonstrated significantly greater blood flow to the true shunt compartment in treated animals. Further studies are required to determine the role of selectins in sepsis-induced lung injury. PMID- 10372633 TI - Experimental study of the effects of splenectomy and partial splenectomy on bacterial translocation. AB - BACKGROUND: The aim of this study is to evaluate the effect of splenectomy and partial splenectomy in a burn-induced bacterial translocation model and to study Kupffer cell (KC) morphology and number. METHODS: Mice were divided into sham burn and burn groups. Each group was also subdivided to sham-splenectomy, partial splenectomy, and splenectomy subgroups. At day 0, operations were performed. At the postoperative 10th day, a sham burn or burn injury was made in all animals. Twenty-four hours later, cultures for bacterial translocation were obtained and livers were evaluated for the quantity and morphology of KCs. RESULTS: Burned splenectomized animals had significantly decreased bacterial translocation when compared with sham-splenectomized animals (p = 0.031). Interestingly, in both the sham-burned and burned groups, splenectomy subgroups had significantly higher numbers of KCs compared with partial-splenectomy and sham-splenectomy subgroups (p<0.00000). Burn injury caused a significant decrease of KC numbers in all subgroups compared with their correspondent sham-burned subgroups (p<0.05). CONCLUSION: Results revealed that splenectomy decreases bacterial translocation and also increases the number of KCs. PMID- 10372634 TI - Trauma registry databases: a comparison of data abstraction, interpretation, and entry at two level I trauma centers. AB - BACKGROUND: A key part of a viable trauma system is the Trauma Registry (TR), used for research, education, and performance improvement. This study sought to assess the consistency of data abstraction, interpretation, and entry by two hospitals with an identical TR database program. METHODS: In phase I, trauma service personnel were queried as to how data were abstracted and entered into the TR. In phase II, a 1-year retrospective review was conducted of TR data for two trauma centers in San Antonio, Texas. RESULTS: The phase I review revealed substantial variances in the coding and abstracting of TR data in 30 of the 500 elements (6%). Phase II demonstrated that, because of these variances, considerable differences resulted in coded types and causes of injury. CONCLUSION: This study illustrates that these variances can impact attempts to combine databases, establish norms, or assess institutional outcomes. To ensure the standardization and accuracy of this valuable information, changes may be required. Recommendations include standardization and education. A uniform trauma registry or national certification may be necessary. PMID- 10372635 TI - Violence in America: a public health crisis--domestic violence. AB - Domestic violence is a major public health problem. It is important that physicians are aware of the extent and pervasiveness of this disease. It is important to identify potential victims of domestic violence when they are encountered in the hospital or office environment. A few, short, carefully asked questions can serve an important surveillance and diagnostic function. Once domestic violence is identified, a well thought out, sensitive, safe plan of action should be discussed with the victim. In this way, not only will the current event be well managed, but also the potential for mitigating further domestic violence events will be initiated. Through this document, EAST hopes to add its voice to that of other physician groups to serve as a catalyst for broad education on the subject of domestic violence as well as activating victim advocacy among physicians and others who come into contact with this problem in their patients. PMID- 10372636 TI - Impact of a dedicated trauma service on the quality and cost of care provided to injured patients at an urban teaching hospital. AB - BACKGROUND: To determine the impact of a dedicated trauma service on cost and quality of care in an urban teaching hospital, a before-and-after study was designed. The key elements of the trauma service were dedicated in-house trauma attending surgeons and residents, and continuity and integration of trauma care. METHODS: Injury Severity Scores and probabilities of survival for each patient were calculated from the hospital International Classification of Diseases, Ninth Revision, codes, and individual patient costs were estimated from charges using the Medicare Cost Report. RESULTS: The trauma service resulted in a significant increase in the severity of injuries. There was a highly significant (p<0.001) increase in the mean probability of death (from 0.16 to 0.21). There was no change in actual mortality. Although the mean cost of care increased by 16.6%, there were small reductions in the cost of care of the most severely injured patients. CONCLUSION: A dedicated trauma service has a positive impact on the quality of care. PMID- 10372637 TI - Blunt carotid artery injuries: difficulties with the diagnosis prior to neurologic event. AB - OBJECTIVE: To evaluate the incidence, timing of diagnosis, clinical factors for adverse outcome, and role of anticoagulant, surgical therapy, or endovascular intervention for patients with blunt carotid artery injury (BCAI). METHODS: Retrospective review of the records of patients who sustained BCAI between 1987 and 1997. RESULTS: There were 18 men and 12 women, with an average age of 29 years. The diagnosis of BCAI was initially suspected in 15 patients after a major or new neurologic event, and in 15 patients after changes were shown by computed tomography. BCAI was confirmed by arteriography in 29 patients and by magnetic resonance angiography in 1 patient. Treatment consisted of antiplatelet therapy (n = 9), anticoagulation (n = 8), surgical repair (n = 6), observation (n = 4), and endovascular embolization (n = 3). With some type of treatment, 14 patients with no neurologic deficits remained stable; however, treatment improved the final neurologic outcome in 8 patients (20%). Three patients remained with severe deficits, and five patients died. CONCLUSION: The consequences of BCAI may be devastating. In our study, there were no reliable means to suspect this injury before neurologic symptoms or abnormalities show on computed tomographic scan. Although external signs are occasionally helpful, most patients have no pattern of injury to suggest BCAI. For patients whose findings after neurologic examination do not correlate with those on the computed tomographic scan, an immediate angiogram is indicated. Occasionally, a proximal injury can be surgically repaired, but in most patients, anticoagulation therapy appears to be the best treatment to avoid or improve neurologic deficits. PMID- 10372638 TI - Natural history of penetrating lower extremity venous injuries: a case report of recanalization after nonoperative treatment. PMID- 10372639 TI - Giant duodenal diverticulum: presentation by blunt trauma. AB - Most duodenal diverticula are asymptomatic, small (i.e., less than 5 cm in greatest dimension), acquired, extraluminal, and false. The only report of a massive or giant duodenal diverticulum (i.e., 10 cm or more), in the current literature, included severe nocturnal diarrhea. The present case report is the incidental finding of a massive duodenal diverticulum in a 34-year-old male trauma victim. The insidious nature of this case and the patient's age suggest a congenital etiology. We believe this is the first report of such a case. PMID- 10372640 TI - Spontaneous rupture of the urinary bladder in the alcoholic patient. AB - A case of a young man with an acute abdominal condition and hematuria is presented. BUN and SCr levels were markedly elevated. Retrograde cystography revealed intraperitoneal extravasation of contrast material. At exploration, a large intraperitoneal bladder perforation was noted and repaired in two layers. Recovery was uneventful. The presentation, diagnosis, and treatment of spontaneous rupture of the urinary bladder are discussed. PMID- 10372641 TI - Retrograde arterial bullet embolus to the coronary artery: case report. PMID- 10372642 TI - Wallstent placement in a renal artery after blunt abdominal trauma. PMID- 10372643 TI - Successful simultaneously stapled lobectomy for a stab wound to the lung with massive hemorrhage: case report. PMID- 10372644 TI - Autoregressive modeling of the EEG in systemic kainic acid-induced epileptogenesis. AB - Background activity as well as three kinds of bilateral epileptiform discharges, recorded from the cerebral cortex and hippocampus of freely behaving rats treated with intravenous kainic acid (KA), were analysed by the directed transfer function (DTF) method within multivariate autoregressive modeling of the EEG. This method reveals statistical influence (flow of activity) between brain regions at different frequencies. There was no significant influence between rhythms in different brain regions in the background EEG. Early after KA administration, low frequency rhythms (< 10Hz) in the frontal cortex began to lead slow rhythms in other areas and high frequency rhythms (20-60 Hz), possibly gamma oscillations, intensified in the hippocampus. In spike-wave discharges, frontal cortex led both low and high frequency rhythms. Initially during generalised non-convulsive discharges, slow rhythms originated from frontal cortex and high frequency rhythms from hippocampus while later, slow rhythms as well, often arose from hippocampus. During the convulsive discharge, the flow of activity of dominant slow rhythms repeatedly changed between hippocampus and neocortex, with more frequent dominance of the hippocampus, while hippocampus continued to lead high frequency rhythms. We conclude that KA-induced epileptiform discharges are cortical and hippocampal events, specifically that the frontal cortex is early to express low frequency rhythms and the hippocampus, high frequency rhythms. More generally, the findings suggest that epileptiform discharges result from interacting rhythms of different frequencies that arise from different structures, and that gamma oscillations possibly contribute to widespread synchronisation during some forms of epileptogenesis. PMID- 10372645 TI - Information metabolism as a model of human experiences. AB - This paper presents Kepinski's concept of "information metabolism", which attempts to describe processing of the information as an analogy of energy metabolism. It is a biological model based upon the structural organization of the cell. The following terms: control center, boundaries, functional structures, centers of elimination and centers of energy are considered here. The model is complementary to computational models, which are helpful in research and theoretical studies, but not useful in interpersonal contacts. The model of information metabolism aims at explanation of relations between psychological and somatic processes and helps in understanding of psychotherapeutic processes. PMID- 10372646 TI - Prediction of brain damage severity: demographic corrections. AB - This study investigated the value of using age and education corrections to the Alternative Impairment Index (AII), to predict the severity of brain damage on the General Neuropsychological Deficit Score (GNDS) and the Halstead Impairment Index (HII). A sample of 40 normal, psychiatric and brain-damaged subjects who had been administered the HRNTB was obtained and the AII, GNDS and HII were calculated for each subject. Hit rates for agreement on level of severity, after correction for age and education effects, for the GNDS and AII were 52.5% and for the AII and HII were 47.5%. Examination of the data suggested that AII, after correction for age and education effects, reflects severity of brain damage at a chance level on both the HII and on the GNDS. PMID- 10372647 TI - Impairments in judgment of chimeric faces by schizophrenic and affective patients. AB - The hypothesis that schizophrenic and affective patients have differential impairments in judgment of facial emotional expressions was tested on 55 right handed patients: 15 in each of two groups of schizophrenic patients, with positive and negative symptoms; and 10 in each of two groups of bipolar affective patients, in manic and depressive states. In addition, 37 normal control subjects were also tested. The subjects were presented with eight schematic drawings of chimeric faces (each depicting a positive emotion in a given hemiface, and a negative emotion in the other hemiface), as well as with two drawings of composite faces (each depicting either a positive or a negative emotion). Subjects judged the emotions depicted by the facial expressions, as well as their intensity. The data, analyzed by analyses of variance, showed that normals judged the chimeric expressions on the basis of the emotions depicted by the left hemifaces. This tendency was weaker among the psychiatric patients. Schizophrenics with negative symptoms judged positive expressions in the left hemifaces as depicting negative emotions, and negative expressions as depicting positive emotions. Schizophrenics with positive symptoms and manic patients judged all expressions as depicting positive emotions. Depressive patients showed a stronger tendency to judge negative expressions as depicting negative emotions than positive expressions as depicting positive emotions. No significant group differences appeared in judgment of composite faces (except for schizophrenic with negative symptoms who were more accurate in judging positive than negative expressions). Patients performances were interpreted in terms of differential dysfunctions in posterior areas of the right cerebral hemisphere which might be associated with bilateral effects of dysfunctions in anterior cerebral areas. PMID- 10372648 TI - Electrocortical responses of headache patients to the simulation of 10 kHz sferics. AB - Many headache patients believe that weather changes act as pain triggers. Therefore, the present study investigated the psychophysiological influence of an indicator of atmospheric instability, Very Low Frequency (VLF)-sferics, on 32 subjectively weather-sensitive women suffering from migraine attacks and/or tension-type headaches. It was analyzed if sferics exposure is able to induce electrocortical changes as well as headache symptoms. The subjects, who had been divided into two groups, participated in a sferics simulation study. The experimental group (n = 16) underwent a ten-minute exposure to 10kHz-sferics impulses followed by 20 minutes without treatment in order to examine possible prolonged sferics effects. The control group (n = 16) received no treatment. As dependent measures, EEG spectral power was compared between the two groups at six electrode sites (F3/F4; P3/P4; O1/O2). Sferics exposure provoked increases in absolute alpha and beta power during the treatment. The alpha power enhancement was still present at parietal sites at the end of registration (20 minutes after the end of exposure). The stimulation did not induce headache symptoms. PMID- 10372649 TI - Treatment with AC pulsed electromagnetic fields improves olfactory function in Parkinson's disease. AB - Olfactory dysfunction is a common symptom of Parkinson's disease (PD). It may manifest in the early stages of the disease and infrequently may even antedate the onset of motor symptoms. The cause of olfactory dysfunction in PD remains unknown. Pathological changes characteristic of PD (i.e., Lewy bodies) have been demonstrated in the olfactory bulb which contains a large population of dopaminergic neurons involved in olfactory information processing. Since dopaminergic drugs do not affect olfactory threshold in PD patients, it has been suggested that olfactory dysfunction in these patients is not dependent on dopamine deficiency. I present two fully medicated Parkinsonian patients with long standing history of olfactory dysfunction in whom recovery of smell occurred during therapeutic transcranial application of AC pulsed electromagnetic fields (EMFs) in the picotesla flux density. In both patients improvement of smell during administration of EMFs occurred in conjunction with recurrent episodes of yawning. The temporal association between recovery of smell and yawning behavior is remarkable since yawning is mediated by activation of a subpopulation of striatal and limbic postsynaptic dopamine D2 receptors induced by increased synaptic dopamine release. A high density of dopamine D2 receptors is present in the olfactory bulb and tract. Degeneration of olfactory dopaminergic neurons may lead to upregulation (i.e., supersensitivity) of postsynaptic dopamine D2 receptors. Presumably, small amounts of dopamine released into the synapses of the olfactory bulb during magnetic stimulation may cause activation of these supersensitive receptors resulting in enhanced sense of smell. Interestingly, in both patients enhancement of smell perception occurred only during administration of EMFs of 7 Hz frequency implying that the release of dopamine and activation of dopamine D2 receptors in the olfactory bulb was partly frequency dependent. In fact, weak magnetic fields have been found to cause interaction with biological systems only within narrow frequency ranges (i.e., frequency windows) and the existence of such frequency ranges has been explained on the basis of the cyclotron resonance model. PMID- 10372650 TI - OPC-13013, a cyclic nucleotide phosphodiesterase type III, inhibitor, inhibits cell proliferation and transdifferentiation of cultured rat hepatic stellate cells. AB - Activated hepatic stellate cells (HSC; lipocytes; Ito cells) proliferate and are responsible for extracellular matrix synthesis during hepatic fibrogenesis. During activation, HSC undergo transdifferentiation into myofibroblasts expressing alpha-smooth muscle actin (alpha-SMA). Adenosine 3', 5'-cyclic monophosphate (cyclic AMP) is an ubiquitous intracellular signaling molecule, and is upregulated by the activation of adenylate cyclase and downregulated via hydrolysis by cyclic nucleotide phosphodiesterases (PDEs). Recently, increased intracellular cyclic AMP has been shown to inhibit HSC activation. The aim of the current study was to determine the effects of inhibition of PDEs on cell proliferation and transdifferentiation in cultured rat HSC. Cell proliferation was determined by [3H]thymidine incorporation, and Western blot analysis was performed for detection of alpha-SMA, a phenotypic marker of transdifferentiation into myofibroblast. When the cells were exposed to 3-isobutyl-1-methylxanthine (IBMX; 50-1000 microM), a nonselective PDE inhibitor, serum-stimulated [3H]thymidine incorporation was suppressed in a dose-dependent manner with a maximum inhibition of 66% at a concentration of 500 microM OPC-13013 (1-60 microM), a selective PDE III isoenzyme inhibitor, induced a dose-dependent inhibitory effect on serum-stimulated DNA synthesis that reached a maximum inhibition of 95% at a concentration of 60 microM, while neither 8-methoxymethyl 3-isobutyl-1-methylxanthine (8-MMX), a PDE I isoenzyme inhibitor, nor Ro-20-1724, a PDE IV isoenzyme inhibitor, had an inhibitory effect. Western blot analysis revealed that IBMX or OPC-13013 decreased alpha-SMA expression, while other selective PDE isoenzyme inhibitors did not have a suppressive effect. IBMX, OPC 13013 or Ro-20-1724, but not 8-MMX augmented forskolin-induced increase in intracellular cyclic AMP levels although cyclic AMP levels were not affected by treatment with any of these PDE inhibitors alone. These data indicate that inhibition of PDEs, especially PDE III isoenzyme, can produce an inhibitory effect on HSC activation. The PDE III isoenzyme may contribute to the regulation of HSC activation during fibrogenesis. In addition, OPC-13013 may have the potential to inhibit initiation and progression of hepatic fibrosis by interfering with HSC activation. PMID- 10372651 TI - Blocking of cell proliferation, cytokines production and genes expression following administration of Chinese herbs in the human mesangial cells. AB - In the hope of identifying agents of therapeutic value in immuoglobulin A nephropathy (IgA-N), we tested crude methanol extracts of 15 Chinese herbs for their effect on human mesangial cell proliferation. The results indicated that 4 out of the 15 crude extracts inhibited human cells proliferation activated by IL 1beta and IL-6. The extracts and their median inhibitory concentrations were as follows (in microg/ml): Ludwiga octovalvis (MLS-052), 49.9 +/- 1.8; Rhus semialata (MLS-053), 31.2 +/- 1.6; Tabernaemontana divaricata (MLS-054), 50.0 +/- 2.1; Amepelopsis brevipedunculata (MLS-059), 42.9 +/- 1.1. These findings indicate that human mesangial cells were most sensitive to MLS-053 treatment. These herbs also decreased interleukin-1beta (IL-1beta) and tumor necrosis factor alpha (TNF-alpha) production. Moreover, IL- 1beta mRNA expression was inhibited by Rhus semialata (R. semialata; MLS-053). It is unlikely that cytotoxicity was involved, because no cell deaths were observable. We hypothesize that the inhibitory mechanisms of these Chinese herbs may be related to the impairments of gene expression and production of cytokines in human mesangial cells. Plans are underway for the isolation of pure compounds from these Chinese herbs and the elucidation of their mechanisms of action. PMID- 10372652 TI - Cocaethylene formation in rat, dog, and human hepatic microsomes. AB - The dog and rat are important animal models for studying the role of cocaethylene in the pharmacodynamic interaction between cocaine and ethanol. In a previous study in our laboratory it was found that a cocaine dose of 3 mg/kg IV and ethanol 1 g/kg IV failed to produce detectable concentrations of cocaethylene in the plasma of dogs. In follow up to this result, the pharmacokinetic disposition of cocaine and cocaethylene in the dog were determined to be similar. These results suggested significant differences between animal and human cocaethylene formation may occur. To test this possibility the in vitro formation of cocaethylene was determined in rat, dog and human hepatic microsomal preparations containing cocaine (0-7 mM) and ethanol (50 mM). Nonlinear least-squares regression was used to estimate Km and Vmax and the results were compared statistically. The mean +/- standard deviation for Km and Vmax in the rat, dog and human were 0.53 +/- 0.04, 0.97 +/- 0.07, and 0.56 +/- 0.08 mM, and 390 +/- 9, 233 +/- 6, and 60 +/- 3 pmol/minute/mg protein, respectively. The Km in the dog was significantly greater (p<0.05) than the Km in the rat and human. The Vmax was statistically different among all three species (rat>dog>human; p<0.05). These results demonstrate that cocaethylene formation is greater in dog than human hepatic microsomes, which is in contrast to in vivo studies that appear to show that humans produce more cocaethylene than dogs. It is suggested by the authors that route of cocaine administration may be an important factor in the formation of cocaethylene when cocaine and ethanol are co-administered. PMID- 10372654 TI - Complex regulation of prothymosin alpha in mammary tumors arising arising in transgenic mice. AB - Expression of prothymosin alpha (PTA) has been related to cell proliferation, both normal and pathological. PTA has also been proposed to be a target of the c myc protooncogene. To study PTA mRNA levels during pathological cell growth, and especially the effect of the activation of specific oncogenes on PTA expression, we have studied its expression in tumors that arise in transgenic mice. We found high PTA levels in mammary tumors arising in c-myc, c-neu, and v-ras transgenic mice. Levels of this protein were variable between different tumors, and there is a differential regulation of PTA respect to other putative c-myc target genes, such as Ornithine Decarboxylase (ODC). Furthermore, expression of PTA is not absolutely dependent of c-myc expression, as shown by MYC depletion experiments performed with antisense oligonucleotides. We conclude that regulation of PTA in these tumors is complex and depends on more than a single activated oncogene. PMID- 10372653 TI - Histological changes and neurotransmitter levels three months following perinatal asphyxia in the rat. AB - The involvement of excitatory amino acids (EAA) in the pathogenesis of hypoxic ischemic states is well-documented. Information on the role of overexcitation by EAA in perinatalasphyxia (PA), however, is limited and data from adult models cannot be directly extrapolated to immature systems. Moreover, most adult models of ischemia are representing stroke rather than PA. We decided to study long term effects in a non-invasive rat model of PA resembling the clinical situation three months following the asphyctic insult. Morphometry on Nissl - stained sections was used to determine neuronal death in frontal cortex, striatum, hippocampus CA1, hypothalamus and cerebellum L1, and the amino acids glutamate, glutamine, aspartate, GABA, taurine, arginine as well as histamine, serotonin and 5-hydroxy indoleacetic acid were determined in several brain regions and areas. Morphometry revealed that neuronal loss was present in the hippocampal area CA1 in all groups with PA and that morphological alterations were significantly higher in the cerebellar granular layer. The prominent light microscopical finding in all areas of asphyctic rats studied was decreased Nissl-staining, suggesting decreased cellular RNA levels. Glutamate, aspartate and glutamine were significantly elevated in the hypothalamus of asphyctic rats probably indicating overstimulation by EAA. Excitotoxicity in this area would be compatible with findings of emotional / behavioral deficits observed in a parallel study in our model of PA. Our observations point to and may help to explain behavioral and emotional deficits in Man with a history of perinatal asphyxia. PMID- 10372655 TI - The priming effect of 12(S)-hydroxyeicosatetraenoic acid on lymphocyte phospholipase D involves specific binding sites. AB - We have previously shown that 12(S)-hydroxyeicosatetraenoic acid (12(S)-HETE) enrichment primed human peripheral blood mononuclear cells for phospholipase D activation by mitogens. Given that 12(S)-HETE-enriched cells stimulated with concanavalin A released free 12(S)-HETE in the extracellular medium, and that the priming effect of 12(S)-HETE on phospholipase D was suppressed by the non permeant drug, suramin, we hypothesized an extracellular mechanism for 12(S)-HETE induced PLD activation. Using [3H]12(S)-HETE as a ligand and a rapid filtration technique, we have pointed out the presence of specific low-affinity 12(S)-HETE binding sites on intact human mononuclear cells and lymphocytes. [3H]12(S)-HETE binding was efficiently displaced by other monohydroxylated and n-3 fatty acids but not by oleate and arachidonate, and was also significantly inhibited by suramin and pertussis toxin. Furthermore, 12(S)-HETE-induced PLD activation was strongly inhibited by pertussis toxin and genistein, but was not PKC-dependent. In addition, 12(S)-HETE also potentiated the ConA-induced tyrosine phosphorylation of a 46-50 kDa protein, which was inhibited by genistein. Collectively, these results suggest that 12(S)-HETE binding sites on human lymphocytes may be coupled to phospholipase D through pertussis toxin sensitive G proteins and tyrosine kinases. PMID- 10372656 TI - Gamma-hydroxybutyrate increases gastric emptying in rats. AB - The influence of gamma-hydroxybutyrate (GHB; 10, 50 or 100 mg/kg orally) and of its receptor antagonist, NCS-382 (25, 100 or 200 mg/kg orally, and 100 or 200 mg/kg intraperitoneally), on gastric emptying was studied in rats by measuring the serum level of acetaminophen (20 mg/rat orally, 30 min after GHB or NCS-382) 15, 30, 45 and 60 min after acetaminophen administration, or the amount of acetaminophen still present in the stomach 30 min after its administration. The highest dose of GHB produced a significant increase in 15 and 30 min serum levels of acetaminophen, indicating an acceleration of gastric emptying. A similar result was obtained with the prokinetic drug cisapride, at the oral dose of 2 mg/kg. On the other hand, NCS-382 significantly and dose-dependently reduced the serum levels of acetaminophen at every time of blood sampling, indicating a delay of gastric emptying, an effect confirmed by the amount of acetaminophen still present in the stomach 30 min after administration. Moreover, NCS-382 antagonized the prokinetic effect of GHB. These results may suggest for GHB (and/or possibly for its metabolites) a role in rat stomach motility. PMID- 10372657 TI - High lovastatin doses combined with hypercholesterolemic diet induce hepatic damage and are lethal to the CD-1 mouse. AB - The addition of 1% lovastatin (LVT) to hypercholesterolemic diets [1% cholesterol or 1% cholesterol plus 0.1% sodium deoxycholate (HD)] induced hepatic damage and was lethal to CD-1 mice in the first days of treatment; the females were more resistant than males. LVT or HD administered alone was harmless to male or female mice. After a 3-day treatment all groups that received LVT (1%, 0.1% or 0.05%) plus HD showed a higher percentage of liver weight, with respect to whole body weight. Cholesterol serum levels increased in males with HD, but remained low in female mice. In the liver, total lipids and cholesterol levels increased in male mice with HD, but cholesterol remained unchanged in females. The addition of LVT to HD prevented the increase of serum and liver cholesterol levels in male mice. These results allow us to propose the CD-1 male-mouse as a model to evaluate the toxicity of LVT or other vastatins. PMID- 10372658 TI - Glucose consumption by rats decreases cytochrome P450 enzyme activity by altering hepatic lipids. AB - Although glucose is a ubiquitous nutrient, increased consumption of glucose decreases the metabolism of numerous drugs in humans and animals. To understand the mechanisms involved that cause decreased drug metabolism in rats that consume glucose in their water, enzyme activity and expression as well as determining the contribution of the lipids toward decreasing in vitro metabolic activity were investigated. Enzyme assays of hepatic CYP1A2, 2C6, 2C11 and 3A2 showed significant decreases in activity from glucose-treated rats compared to control. While immunodetection of CYP1A1, 2B1/2, 2C11, and 3A1/2 showed no significant difference in protein expression. Hepatic fatty acid synthase activity increased in the glucose-treated rats compared to controls. Studies with glucose-treated microsomal lipids reconstituted with microsomal proteins from control rats caused a significant decrease in benzyloxyresorufin O-dealkylase activity. The results presented here support the hypothesis that the activities of cytochrome P450 proteins are altered by modulating their catalytic activity as a result of the lipid environment rather than changing the level of expression of the individual enzymes. PMID- 10372659 TI - NGF-differentiated and undifferentiated PC12 cells vary in induction of apoptosis by ethanol. AB - The present study was undertaken to determine whether the neurotoxic effects of ethanol vary between undifferentiated and differentiated neurons. For this study, untreated rat pheochromocytoma (PC12) cells and PC12 cells treated for 8-10 days with nerve growth factor (NGF) were used as models of undifferentiated and differentiated neurons, respectively. Treatment of differentiated PC12 cells with 150 mM ethanol resulted in a loss of cells whereas a similar treatment of undifferentiated cells had no effect. In contrast, 50 mM ethanol enhanced apoptosis initiated by serum withdrawal in undifferentiated cells while a similar response in the differentiated cells required 150 mM ethanol. This study demonstrates that undifferentiated and differentiated neuronal cells differ in their sensitivity to the neurotoxic actions of ethanol. PMID- 10372660 TI - The nature of prooxidant activity of vitamin C. AB - It was recently reported that vitamin C (500 mg/day for 6 weeks) administered as a dietary supplement to healthy humans exhibits a prooxidant, as well as an antioxidant effect in vivo. Here we show that high intakes of vitamin C (500 mg/kg b.w. for 4 days) in the rat are able to markedly induce hepatic cytochrome P4502E1-linked monooxygenases, measured as p-nitrophenol hydroxylase activity and corroborated by means of Western blot analyses. Furthermore, using Electron Paramagnetic Resonance Spectroscopy (EPR) coupled to a spin-trapping technique, we have also found that this induction generates large amounts of the anion radical superoxide (O2-). Therefore we can conclude that the adverse prooxidant outcomes (i.e. oxidative DNA damage) associated to vitamin C supplementation, being associated to a typical reversible boosting effect (i.e. enzymatic induction), may be easily controlled by a discontinuous supply. However, since the induced P4502E1 isoforms by vitamin C are responsible for ethanol metabolism to highly reactive radicals, care should be taken even in moderate drinkers. PMID- 10372661 TI - Age dependent response to exogenous estrogen on micturition, contractility and cholinergic receptors of the rat bladder. AB - The age related effects of 17beta-estradiol (E) supplementation on micturition and contractility of ovariectomized rats (OVX) were evaluated. Studies were carried out in young, 2 month, and mature, 10 month old rats which were distributed into three groups: Sham-operated (SHAM), (OVX), and (OVX+E). Following treatment, urodynamic studies were performed followed by an in vitro bladder tissue evaluation. Urodynamic studies show age and time related changes in bladder function. The in vitro results show that the hormone deprived tissues of 2 months old rats had a decreased responsiveness to cholinergic stimulation; maximum contractile force occurred at 78% and 187% for the SHAM. The response from the OVX+E tissues was evident at 113%. E supplementation of the mature rats increased bladder contractile force to the same levels as SHAM (156% and 176%). The response of the mature OVX rats remained significantly below that of SHAM or OVX+E rats. Findings suggest that the impact of E on bladder function depends on age at which it is given. Differential response between young and mature to exogenous E indicates that endogenous estrogen plays a major role in the neuromuscular development of normal bladder function and micturition reflexes. Contractility data show that OVX in young rats irreversibly decreases the response of the bladder to cholinergic stimulation, suggesting that exogenous E partially restores function while in mature rats, exogenous E was able to reverse the effects of OVX. PMID- 10372662 TI - Clinical review 105: Stress and the reproductive cycle. PMID- 10372663 TI - Clinical review 106: Amenorrheic bone loss. PMID- 10372664 TI - Clinical review 107: Role of gonadal steroids in the sexual dimorphisms in body composition and circulating concentrations of leptin. PMID- 10372665 TI - Clinical review 108: The molecular and neuroanatomical basis for estrogen effects in the central nervous system. PMID- 10372666 TI - Therapeutic controversy: Hormone replacement therapy--where are we going? PMID- 10372667 TI - Therapeutic controversy: Screening for postpartum thyroiditis. PMID- 10372668 TI - Therapeutic controversy: The safety of third-generation oral contraceptives. PMID- 10372669 TI - Women and Alzheimer's disease. AB - AD is a major public health problem, and demographic trends have led to its being called the epidemic of the century. Because of increased longevity and the special challenges of ERT, women are well placed to both be at risk for and the beneficiaries of advances in AD therapy. Overall increases in health consciousness may impact future AD risk, and it is encouraging that women frequently outnumber men in clinical trials of new therapeutic agents in AD. The risk of AD from environmental exposures in the overall life experiences of women is unclear. To the extent that education and work promote the development of brain areas such as the association cortex that are preferentially affected in AD, creating a neuronal reserve, advances in women's overall place in society may further help protect them from the ravages of AD. PMID- 10372670 TI - Cardiovascular disease in the diabetic woman. AB - The incidence of cardiovascular disease (CVD) with age is increasing in the United States, and elderly women constitute a disproportional component of the aging population. Elderly women also have a relatively high incidence of diabetes, which contributes to this relatively high CVD risk. Although CVD is less common in premenopausal women than in men, this difference begins to disappear after the onset of menopause, presumably related to decreased levels of female sex hormones (estrogen and/or progesterone). Diabetes mellitus removes the normal premenopausal gender-related differences in the prevalence of CVD by mechanisms that are not clearly defined, including metabolic and hemodynamic factors associated with diabetes. Dyslipidemia in diabetes mellitus consists of low high density lipoprotein cholesterol, elevated triglyceride levels, and a small, dense, more atherogenic low density lipoprotein particle (i.e. oxidized). Dyslipidemia interacts with associated hemodynamic (i.e. hypertension) and metabolic abnormalities (i.e. increased platelet aggregation and plasminogen activator inhibitor-1 levels) to promote CVD risks in diabetic women. Recent controlled trials underscore the critical importance of aggressively treating CVD risk factors, especially dyslipidemia, in women with diabetes. PMID- 10372671 TI - Adolescent nutrition in the prevention of postmenopausal osteoporosis. PMID- 10372672 TI - What causes low rates of child-bearing in congenital adrenal hyperplasia? PMID- 10372673 TI - Hormones and heart disease in women: Heart and Estrogen/Progestin Replacement Study in perspective. AB - Despite the nearly universal finding from observational studies that postmenopausal estrogen therapy reduces the risk of CHD and the multiple plausible mechanisms by which estrogen might reduce the risk of CHD, hormone therapy had no benefit in the only large randomized clinical trial to date. Although it is possible that estrogen taken over the long term actually reduces CHD risk, it is not reasonable to begin the regimen used in HERS to prevent new or recurrent heart disease, given the observed excess early risk. Given the possible long term benefit, women who are already taking hormone replacement therapy may elect to remain on it. Women who are undecided should be asked to consider participation in clinical trials. The HERS has dramatically illustrated the need for them. PMID- 10372674 TI - Gestational diabetes: risk or myth? AB - As currently defined (5), gestational diabetes is associated with important perinatal and long-term health risks. Many of the risks increase, in relation to the severity of maternal hyperglycemia. For perinatal risks to infants, the relationship seems to be continuous. Maternal fasting glucose levels can be used to identify subsets of patients with very low and very high risks. The majority of pregnancies lie between these two extremes; and nonglucose measures, such as fetal ultrasound, can be used to enhance risk assessment, thereby minimizing over and undertreatment of patients. The major long-term maternal risk is development of type 1 or type 2 diabetes, predominantly the latter. The risk increases continuously, in relation to maternal glycemia during and, especially, after pregnancy. Patients seem to have a B-cell defect that is characterized by maladaptation to insulin resistance. The B-cell defect is predictive of future diabetes, supporting the testing and clinical application of interventions that minimize insulin resistance to delay or prevent diabetes. Women with impaired glucose tolerance in the first few months postpartum are at particularly high risk for diabetes and should receive the most intensive education, intervention, and follow-up. Offspring of women with GDM are at increased risk for obesity and have an unexpectedly high prevalence of elevated glucose levels during childhood and adolescence. Both genetic and intrauterine environmental influences are likely to contribute to these abnormalities. Optimal strategies to detect and prevent the long-term risks to offspring remain to be established. PMID- 10372675 TI - Hypertension in pregnancy: a potential window into long-term cardiovascular risk in women. PMID- 10372676 TI - Hypertension in women. AB - Hypertension is an important risk factor for cardiovascular disease in women. Although younger, premenopausal women have lower blood pressures than age-matched men, population blood pressure rises with age, and the prevalence of hypertension is higher in older women. Oral contraceptive use increases the risk of hypertension in women, and women using this therapy should have blood pressure monitored twice yearly. The risk of hypertension is low in normotensive women receiving HRT. Few studies of HRT in hypertensive women have been performed, and more information is needed to assess the risk of worsening hypertension in hypertensive postmenopausal receiving this therapy. Investigations of gender differences in pathophysiology and response to treatment of essential hypertension have not been extensive, and current evidence does not support gender-specific treatment of hypertension at the present time. PMID- 10372677 TI - Bone densitometry: the best way to detect osteoporosis and to monitor therapy. AB - The revolution in the field of osteoporosis has been aided and abetted by the advent of bone mass measurement technologies. As they become more widely applicable and more affordable, it is evident that we have the potential to discover the millions of individuals at risk for or with the disease. With effective therapies at hand, it is now possible to prevent and treat osteoporosis. There is every reason, therefore, to apply bone mass measurements as widely as possible to discover those subjects at risk for osteoporosis in a manner that is effective and affordable. PMID- 10372678 TI - Does estrogen adequately protect postmenopausal women against osteoporosis: an iconoclastic perspective. AB - In sum, the skeletal benefits provided by hormone replacement therapy are important, and estrogen should be justifiably considered one of the fundamentals of menopausal management. However, its efficacy in the prevention of osteoporotic fractures should not be overstated. Low bone mass and fractures remain serious threats in older postmenopausal women, even in the presence of hormone replacement. With the recognition of that reality comes a variety of challenges to clinicians and investigators. Clinicians should use estrogen to its best advantage, though, at the same time, remain vigilant of its limitations. Older postmenopausal women who are, or who have been, taking estrogen should not be a priori considered adequately protected against fracture. The same careful clinical evaluation recommended for the protection of those at increased fracture risk, including bone mass measures, should be available to estrogen-taking women in the later postmenopausal period. Recognizing this need, the National Osteoporosis Foundation has recently recommended bone mineral density testing in older women, regardless of estrogen status, and HCFA reimbursement has become available for this indication. In the presence of low bone density, estrogen taking women should be afforded appropriate levels of diagnostic and therapeutic attention, with the intent to further reduce fracture risk. Once estrogen therapy has been elected, patients and their clinicians should be aware that bone loss can still be expected in the later postmenopausal years. Periodic reevaluations of bone density and other risks for fracture (e.g. falls) may be appropriate. In the face of continued good health, those reevaluations can be infrequent, but women who have medical conditions associated with adverse skeletal effects should be followed more closely despite their estrogen therapy. If we are to build on the value of estrogen to improve our approach to osteoporosis, additional data are needed. When fracture risk has been considered in complex models that adjust for account other medical and lifestyle variables, a greater protective effect of estrogen has emerged (12), suggesting that there are factors whose actions attenuate those of estrogen. Prominent candidates include genetics, tobacco and alcohol use, medications, body composition, and propensity to fall. There are undoubtedly others yet unidentified. These should be further defined, and the basic mechanisms for interactions between them and estrogen should be examined. It would be clinically advantageous to use these factors to accurately identify not only the women who would benefit from estrogen replacement but also those who would not. Because estrogen has important beneficial effects, an essential research objective should be the development of therapeutic strategies that combine the advantages of estrogen with other modalities (pharmacological or otherwise), to maximally protect the many estrogen-taking women who may be considered at continued risk for fracture. To whit, the initial reports of combination therapy with bisphosphonates are encouraging (13, 14). Other important skeletal agents can be anticipated to interact with estrogen in as-of yet unforeseen ways. On another front, it can be anticipated that selective estrogen receptor modulators will be subject to estrogen-like limitations when used for osteoporosis prevention/therapy, and their effects will be influenced by a similar, but distinct, array of covariates. Our vision of the role of hormone replacement in the reduction of osteoporotic fracture risk should be considerably sharpened. At one time, estrogen was the lone best hope for the avoidance of fracture, but no longer should we be satisfied with that imperfect solution. The development of much more efficient and effective methods for osteoporosis risk assessment, prevention, and treatment now provides a chance to surpass previous expectations. (ABSTRACT TRUNCATED) PMID- 10372679 TI - Relative versus attributable risk of breast cancer from estrogen replacement therapy. PMID- 10372680 TI - Testing for hereditary cancer risk: Pandora or Prometheus? PMID- 10372681 TI - Androgen replacement in women: a commentary. AB - There is increasing evidence to suggest that many postmenopausal women experience symptoms alleviated by androgen therapy and that such symptoms may be secondary to androgen deficiency. Affected women complain of fatigue, low libido, and diminished well-being, symptoms easily and frequently attributed to psychosocial and environmental factors. When such symptoms occur in the setting of low circulating bioavailable testosterone, testosterone replacement results in significant improvement in symptomatology and, hence, quality of life for the majority of women. Whether the apparent therapeutic effects of testosterone replacement are mediated by testosterone and its metabolite 5alpha- dihydrotestosterone or are a consequence of aromatization to estrogen is not known. Despite the paucity of data regarding its effects, inclusion of testosterone in postmenopausal hormone replacement regimens is not uncommon and is likely to become more widespread with the availability of preparations developed specifically for women. Other novel and even more controversial potential indications for androgen therapy in women are currently being evaluated. These include use in women with premature ovarian failure, premenopausal androgen deficiency symptoms, postmenopausal and glucocorticosteroid-related bone loss, alleviation of wasting syndrome secondary to human immunodeficiency virus infection, and management of premenstrual syndrome. The aim of this commentary is to very briefly review the rationale for the use of testosterone in women, create awareness of some of the therapeutic options available in various countries, and stimulate discussion of this important aspect of women's health. PMID- 10372682 TI - Health issues for women athletes: exercise-induced amenorrhea. PMID- 10372683 TI - Polycystic ovary syndrome (PCOS): arguably the most common endocrinopathy is associated with significant morbidity in women. PMID- 10372684 TI - The individualized approach to menopause management. PMID- 10372685 TI - Neuroendocrine abnormalities in hypothalamic amenorrhea: spectrum, stability, and response to neurotransmitter modulation. AB - To characterize the neuroendocrine patterns of abnormal GnRH secretion in hypothalamic amenorrhea (HA), 49 women with primary and secondary HA underwent frequent sampling of LH in a total of 72 baseline studies over 12-24 h. A subset of women participated in more than one study to address 1) the variability of LH pulse patterns over time; and 2) the impact of modulating opioid, dopaminergic, and adrenergic tone on LH secretory patterns. The frequency and amplitude of LH secretion was compared with that seen in the early follicular phase (EFP) of normally cycling women. The spectrum of abnormalities of LH pulses was 8% apulsatile, 27% low frequency/low amplitude, 8% low amplitude/normal frequency, 43% low frequency/normal amplitude, 14% normal frequency/normal amplitude. Of patients studied overnight, 45% demonstrated a pubertal pattern of augmented LH secretion during sleep. Of patients studied repeatedly, 75% demonstrated at least 2 different patterns of LH secretion, and 33% reverted at least once to a normal pattern of secretion. An increase in LH pulse frequency was seen in 12 of 15 subjects in response to naloxone (opioid receptor antagonist). Clonidine (alpha-2 adrenergic agonist) was associated with a decrease in mean LH in 3 of 3 subjects. An increase in LH pulse frequency was seen in 4 of 8 subjects in response to metoclopramide (dopamine receptor antagonist), but the response was not statistically significant. Baseline abnormalities in LH secretion did not appear to influence response to neurotransmitter modulation. CONCLUSIONS: 1) HA represents a spectrum of disordered GnRH secretion that can vary over time; 2) LH pulse patterns at baseline do not appear to influence the ability to respond to neurotransmitter modulation; 3) Opioid and adrenergic tone appear to influence the hypothalamic GnRH pulse generator in some individuals with HA. PMID- 10372686 TI - The Reproductive Endocrine Associates of the Massachusetts General Hospital: fifteen years of integrated clinical practice and investigation. PMID- 10372687 TI - Bone loss is correlated to the severity of growth hormone deficiency in adult patients with hypopituitarism. AB - Reduced bone mineral density (BMD) has been reported in patients with isolated GH deficiency (GHD) or with multiple pituitary hormone deficiencies (MPHD). To investigate whether the severity of GHD was correlated with the degree of bone mass and turnover impairment, we evaluated BMD at the lumbar spine and femoral neck; circulating insulin-like growth factor I (IGF-I), IGF-binding protein-3 (IGFBP-3), and osteocalcin levels, and urinary cross-linked N-telopeptides of type I collagen (Ntx) levels in 101 adult hypopituitary patients and 35 sex- and age-matched healthy subjects. On the basis of the GH response to arginine plus GHRH (ARG+/-GHRH), patients were subdivided into 4 groups: group 1 included 41 patients with a GH peak below 3 microg/L (0.9 +/- 0.08 microg/L), defined as very severe GHD; group 2 included 25 patients with a GH peak between 3.1-9 microg/L (4.7 +/- 0.4 microg/L), defined as severe GHD; group 3 included 18 patients with a GH peak between 9.1-16.5 microg/L (11.0 +/- 0.3 microg/L), defined as partial GHD; and group 4 included 17 patients with a GH peak above 16.5 microg/L (28.3 +/ 4.3 microg/L), defined as non-GHD. In all 35 controls (group 5), the GH response after ARG+/-GHRH was above 16.5 microg/L (40.7 +/- 2.2 microg/L). In patients in group 1, circulating IGF-I (P < 0.001), IGFBP-3 (P < 0.05), osteocalcin (P < 0.001), and urinary Ntx levels (P < 0.001) were lower than those in group 3-5, which were not different from each other; the t score at the lumbar spine (-1.99 +/- 0.2) and that at the femoral neck (-1.86 +/- 0.3) were lower than those in groups 3 (-0.5 +/- 0.7, P < 0.01 and -0.3 +/- 0.7, P < 0.01, respectively), 4 ( 0.5 +/- 0.2, P < 0.01 and -0.3 +/- 0.7, P < 0.01, respectively), and 5 (-0.5 +/- 0.2, P < 0.001 and 0.0 +/- 0.02, P < 0.001, respectively). In patients in group 2, circulating IGF-I and IGFBP-3 levels were not different from those in group 1, whereas the t scores at the lumbar spine (-1.22 +/- 0.3) and femoral neck (-0.9 +/- 0.3) were significantly higher and lower, respectively, than those in groups 1 and 5 (P < 0.05) but not those in groups 3 and 4, and serum osteocalcin and urinary Ntx levels were significant higher than those in group 1 and lower than those in groups 3-5 (P < 0.001). To evaluate the effect of isolated GHD vs. MPHD, patients were subdivided according to the number of their hormonal deficits, such as panhypopituitarism with (10 patients) or without (31 patients) diabetes insipidus, GHD with 1 or more additional pituitary deficit(s) (36 patients), isolated GHD (7 patients), 1-2 pituitary hormone deficit(s) without GHD (10 patients), and normal anterior pituitary function (7 patients). The t score at the lumbar spine and femoral neck and the biochemical parameters of bone turnover were not significantly different among the different subgroups with similar GH secretions. A significant correlation was found between the GH peak after ARG+GHRH and IGF-I, osteocalcin, urinary Ntx levels, and the t score at the lumbar spine, but not that at the femoral neck level. A significant correlation was also found between plasma IGF-I levels and the t score at the lumbar spine and femoral neck, serum osteocalcin, and urinary Ntx. Multiple correlation analysis revealed that the t score at the lumbar spine, but not that at the femoral neck, was more strongly predicted by plasma IGF-I levels (t = 3.376; P < 0.005) than by the GH peak after ARG+GHRH (t = -0.968; P = 0.338). In conclusion, a significant reduction of BMD associated with abnormalities of bone turnover parameters was found only in patients with very severe or severe GHD, whereas normal BMD values were found in non-GHD hypopituitary patients. These abnormalities were consistently present in all patients with GHD regardless of the presence of additional hormone deficits, suggesting that GHD plays a central role in the development of osteopenia in hypopituitary patients. PMID- 10372688 TI - Endocrine and metabolic evaluation of human immunodeficiency virus-infected patients with evidence of protease inhibitor-associated lipodystrophy. AB - Multidrug antiretroviral regimens that include human immunodeficiency virus-1 (HIV-1) protease inhibitors are associated with distinct lipodystrophy, hypertriglyceridemia, hyperinsulinemia, and deposition of visceral abdominal adipose tissue. To determine whether these findings are related to abnormalities of adrenal function, we compared the hypothalamic-pituitary-adrenal axes of HIV positive patients who had evidence of protease inhibitor-associated lipodystrophy (PIAL), control volunteers (CON), and patients with Cushing's syndrome (CS). To elucidate the metabolic consequences of the observed lipodystrophy, we measured basal serum lipids and compared glucose and insulin concentrations during an oral glucose tolerance test. Spontaneous plasma cortisol showed normal diurnal variation in PIAL. Cortisol levels were similar in CON and PIAL, and levels in these groups were less than those in CS at all times of the night or day (P < 0.005). Ovine CRH-stimulated morning plasma cortisol levels were similar in PIAL and CON. ACTH was significantly greater in PIAL than CON (P < 0.05) at 0, 15, and 30 min after CRH stimulation. Urinary free cortisol in PIAL (mean +/- SD, 76 +/- 51 nmol/day) was significant lower than those in CON (165 +/- 64 nmol/day; P < 0.001) and CS (1715 +/- 1203 nmol/day; P < 0.001). However, 17 hydroxycorticosteroid excretion was significantly greater in PIAL (43 +/- 23 micromol/day) than in CON (17 +/- 8 micromol/day; P < 0.001), although lower than that in CS (74 +/- 47 micromol/day; P < 0.01). Scatchard analysis revealed normal glucocorticoid receptor number and affinity in PIAL. Serum triglycerides were significantly greater in PIAL (6.57 +/- 5.63 mmol/L) than in CS (1.78 +/- 0.83 mmol/L; P < 0.001) or CON (1.36 +/- 0.84 mmol/L; P < 0.001). Although triglyceride levels were significantly correlated with body mass index for CON and CS, these were not correlated for PIAL. During an oral glucose tolerance test, similar glucose and insulin values were found in PIAL and CS that were greater (P < 0.05) than CON values at 30, 60, 90, and 120 min. We conclude that the lipodystrophy associated with use of HIV-1 protease inhibitors is a syndrome of increased intraabdominal adiposity with concomitant dyslipidemia and insulin resistance, but without total body weight gain and is distinct from any known form of hypercortisolism. Although urinary cortisol disposition seems to be altered in HIV-infected patients who are being treated with multidrug regimens that include protease inhibitors, the decreased free cortisol and increased 17 hydroxycorticosteroid excretion appear to be unlikely explanations for the observed lipodystrophy. The cause remains to be elucidated. PMID- 10372689 TI - Fasting hyperinsulinemia and changes in regional body composition in human immunodeficiency virus-infected women. AB - A novel lipodystrophy syndrome (characterized by insulin resistance, hypertriglyceridemia, and fat redistribution) has recently been described in human immunodeficiency virus (HIV)-infected patients. However, investigation of the lipodystrophy syndrome has generally been limited to men; and a comprehensive evaluation of insulin, lipids, and regional body composition has not been performed in the expanding population of HIV-infected women. In this study, we assessed fasting insulin, lipid levels, virologic parameters, and regional body composition, using dual-energy x-ray absorptiometry, in a cohort of 75 HIV infected women (age, 25-46 yr), in comparison with 30 healthy weight-matched premenopausal control subjects. HIV-infected women demonstrated significant truncal adiposity (38.5 +/- 0.9 vs. 34.9 +/- 1.3%, P < 0.05) hyperinsulinemia (15.9 +/- 1.5 vs. 7.5 +/- 0.6 microU/mL, P < 0.001) and an increased insulin-to glucose ratio (0.2 +/- 0.02 vs. 0.1 +/- 0.03, P < 0.001), compared with control subjects. Insulin and the insulin-to-glucose ratio were increased, even among HIV infected patients with low body weight (<90% of ideal body weight) (insulin, 13.3 +/- 2.8 microU/mL, P < 0.01 vs. control; insulin/glucose, 0.2 +/- 0.04, P < 0.01 vs. control). Insulin and the insulin-to-glucose ratio were most significantly elevated among patients with increased truncal adiposity (insulin, 28.2 +/- 3.2 microU/mL, P < 0.001 vs. control; insulin/ glucose, 0.32 +/- 0.04, P < 0.001 vs. control). In contrast, no differences in insulin were seen in relation to protease inhibitor (PI) use. Similarly, HIV-infected women also demonstrated significant hypertriglyceridemia (144 +/- 15 vs. 66 +/- 23 mg/dL, P < 0.01 vs. controls), which was present even among low-weight patients (148 +/- 32 mg/dL, P < 0.001 vs. control) but was not related to truncal adiposity or PI usage. These data demonstrate significant hyperinsulinemia and truncal adiposity in HIV infected women. Our data suggest that these metabolic abnormalities occur at baseline in HIV-infected women, independent of PI use. However, these data do not rule out a direct effect of PI therapy on fat metabolism or indirect effects of PI therapy to further worsen glucose and lipid homeostasis in association with weight gain and disease recovery. PMID- 10372690 TI - Disruption of the pulsatile and entropic modes of insulin release during an unvarying glucose stimulus in elderly individuals. AB - Insulin is secreted in a pulsatile fashion with measurable orderliness (low entropy). Aging is characterized by alterations in pulsatile insulin release in the fasting state. We undertook the current studies to determine whether disruptions in pulsatile insulin release in response to sustained glucose infusion also accompany the age-related changes in carbohydrate metabolism. Healthy young (n = 10; body mass index, 23 +/- 1 kg/m2; age, 23 +/- 1 yr) and old (n = 10; body mass index, 24 +/- 1 kg/m2; age, 80 +/- 2 yr) volunteers underwent a 600-min hyperglycemic glucose clamp. During the entire 600 min, insulin was sampled every 10 min, and insulin release was evaluated by Cluster analysis. From 240-360 min, insulin was sampled every 1 min, and secretory pulse analysis was conducted using a multiparameter deconvolution technique. During the 1-min sampling interval, basal insulin secretion (P < 0.01), insulin production rate (P < 0.01), pulsatile mean and integrated insulin concentration (P < 0.01), insulin secretory burst mass (P < 0.01), and burst amplitude (P < 0.05) were reduced in the elderly. In addition, interpulse interval was increased in the aged (P < 0.05). In the 600-min studies, interpulse interval was greater in the aged (P < 0.01) and burst number (P < 0.01), basal concentration (P < 0.01), and burst increment (P < 0.05) were less. Approximate entropy, a measure of irregularity of insulin release, was increased in the aged, signifying the loss of orderliness of insulin secretion (P < 0.05). We conclude that in response to a sustained (10-h) glucose infusion, normal aging is characterized by a reduction in mass and amplitude of rapid insulin pulses and a decrease in the frequency, amplitude, and regularity of ultradian pulses. Whether these changes in insulin pulsatility contribute directly to the age-related changes in carbohydrate metabolism will require further clinical studies. PMID- 10372691 TI - Differential hypothalamic-pituitary-adrenal axis reactivity to psychological and physical stress. AB - Healthy men exhibit a differential hypothalamic-pituitary-adrenal axis (HPA) response to exercise stress and fall into two groups: high responders (HR) and low responders (LR). The present study examined whether HR to physical stress also exhibit higher HPA reactivity to psychological stress than LR. We examined 14 HR and 13 LR classified based on their ACTH responses to high intensity exercise after pretreatment with dexamethasone. Both groups were of similar age, height, weight, and fitness level. Trait anxiety scores on the Spielberger Trait Anxiety Scale were not different. Subjects underwent a psychological stress test consisting of an interview and mental arithmetic. This test raised heart rate, blood pressure, and plasma ACTH and cortisol levels in both HR and LR. HR tended to have higher heart rates and blood pressures in anticipation of the psychological stress test than LR. ACTH responses of HR were higher, although not significantly, throughout the psychological stress test than LR. HR had a significantly (P < 0.05) greater net integrated cortisol response to the psychological stress than LR. This suggests that the adrenal cortexes of the HR are hypertropic and/or hypersensitive to ACTH. We conclude that men who are highly responsive to exercise stress are also highly responsive to psychological stress. PMID- 10372692 TI - High serum leptin concentrations during catch-up growth of children born with intrauterine growth retardation. AB - The aim of the study was to investigate how leptin could be involved in catch-up growth of children born with intrauterine growth retardation (IUGR). The study population was made up of 70 newborns with IUGR longitudinally studied during the first 2 yr of life and 35 newborns and 32, 66, and 61 children with normal birth weight aged 3 days, 12 months, and 24 months, respectively. Postnatal patterns of body mass index (BMI) were similar in the 2 groups, but BMI remained significantly lower in IUGR over the study period. In contrast, children born with IUGR aged 1 yr had significantly higher serum leptin levels than normal children (P < 0.0001) independently of BMI. The correlation observed between BMI and serum leptin at birth in both groups and in the control group thereafter disappeared in children born with IUGR. Similarly, sexual dimorphism observed in normal children over the study period was not observed in the IUGR group during the first 2 yr of life. In summary, serum leptin is effective and regulated during the first years of life as it is in older children. Children born with IUGR demonstrate high serum leptin values during the first year of life, with a loss of the regulatory effect of BMI and gender. We suggest that these children develop an adaptative leptin resistance beneficial for their catch-up growth. An alternative hypothesis is that these observations could reflect an adipocyte dysfunction, a consequence of the special time course of adipose tissue development in children born with IUGR. PMID- 10372693 TI - Thickened pituitary stalk on magnetic resonance imaging in children with central diabetes insipidus. AB - Magnetic resonance imaging (MRI) has revealed isolated pituitary stalk (PS) thickening (PST) in certain cases of idiopathic or secondary central diabetes insipidus (DI) due to infiltrative processes. Twenty-six children with DI and PST underwent cerebral MRI at the age of 8 +/- 4 yr and were followed (n = 24) by clinical and MRI evaluation, respectively, for 5.5 +/- 3.6 and 3.0 +/- 2 yr in the absence of any treatment other than hormonal substitutive therapy. Patients were subdivided into groups according to the etiology of the DI: germinoma (n = 4), Langerhans' histiocytosis (n = 5), or idiopathic DI with PST (n = 17). Complete anterior pituitary evaluation for 24 of the 26 patients revealed those suffering from associated GH deficiency (n = 14; with germinoma, n = 1; histiocytosis, n = 3; idiopathic, n = 10) and from multiple hormone deficiencies (n = 7; with germinoma, n = 3; histiocytosis, n = 1; idiopathic, n = 3). At the first MRI evaluation, PS enlargement varied from 2.2-9.0 mm at a proximal (n = 10), distal (n = 2), or middle (n = 6) PS level or along the entire PS (n = 8). The intrasellar content, which usually reflects the anterior pituitary gland, was normal (n = 12), small (n = 8), or enlarged (n = 6). At the last evaluation, a change in MRI features was found in 16 patients; morphological and/or signal changes in the PST (n = 16, of whom 6 showed an increase in PST) and changes in anterior pituitary gland size (n = 8; increased, n = 3; decreased, n = 5) were noted. The presence of a growing suprasellar mass with progressively enlarging PS was demonstrated in the 6 patients who had shown increased PS enlargement 1.8 +/- 1.6 yr after the first MRI. For 4 of them, a diagnosis of germinoma was made 1.3 +/- 0.6 yr after PST identification by MRI performed after the onset of DI, but the other 2 patients showing a suprasellar mass were still categorized as idiopathic at the final clinical evaluation performed 7.8 and 12.3 yr, respectively, after DI onset. In 10 patients (all but 1 with Langerhans histiocytosis, showing idiopathic DI), the PS enlargement was diminished after 2.0 +/- 1.9 yr of MRI follow-up, with a complete reversal of PS enlargement for 5 of them. Suprasellar mass invasion of the PS was related to multiple hormone deficiency. Although intrasellar content enlargement was observed in most patients with germinoma, a normal or small anterior pituitary gland showed no clear relationship to either clinical histories or laboratory values. In conclusion, the natural history of idiopathic isolated central DI with PST is unpredictable. Although germinoma should always be considered during the first 3 yr of follow-up in patients showing isolated DI with PST requiring repeated investigations every 3-6 months, it remains a less frequent etiology for 15% of the cases. PMID- 10372694 TI - Recent status in the occurrence of leukemia in growth hormone-treated patients in Japan. GH Treatment Study Committee of the Foundation for Growth Science, Japan. AB - The Foundation for Growth Science in Japan has monitored the safety and efficacy of GH treatment in GH-deficient patients since 1975. Data were collected from more than 32,000 patients up to December 31, 1997. New leukemia was observed in 14 patients and myelodysplastic syndrome (MDS) in one patient. The types of leukemia were acute lymphocytic leukemia (n = 6; 40%), acute myelocytic leukemia or MDS (n = 7; 47%), and chronic myelocytic leukemia (n = 2; 13%). Leukemia developed in 9 patients during GH treatment and in 6 after the cessation of GH treatment. Six patients had known risk factors for leukemia, such as Fanconi's anemia and previous radiation or chemotherapy. Patient-years of GH therapy was defined as the time from the first dose of GH to the date of the last visit during GH therapy, and patient-years of risk was defined as the time from the first dose of GH to December 31, 1997. The incidence of leukemia of patient-years of GH therapy and patient-years of risk in GH-treated patients without risk factors was 3.0/100,000 and 3.9/100,000, respectively, a figure similar to the incidence in the general population aged 0-15 yr. We conclude that the incidence of leukemia in GH-treated patients without risk factors is not greater than that in the general population aged 0-15 yr, and a possible increased occurrence of leukemia with GH treatment appears to be limited to patients with risk factors. PMID- 10372695 TI - Effect of testosterone treatment on bone mineral density in men over 65 years of age. AB - As men age, their serum testosterone concentrations decrease, as do their bone densities. Because bone density is also low in hypogonadal men, we hypothesized that increasing the serum testosterone concentrations of men over 65 yr to those found in young men would increase their bone densities. We randomized 108 men over 65 yr of age to wear either a testosterone patch or a placebo patch double blindly for 36 months. We measured bone mineral density by dual energy x-ray absorptiometry before and during treatment. Ninety-six men completed the entire 36-month protocol. The mean serum testosterone concentration in the men treated with testosterone increased from 367 +/- 79 ng/dL (+/-SD; 12.7 +/- 2.7 nmol/L) before treatment to 625 +/- 249 ng/dL (21.7 +/- 8.6 nmol/L; P < 0.001) at 6 months of treatment and remained at that level for the duration of the study. The mean bone mineral density of the lumbar spine increased (P < 0.001) in both the placebo-treated (2.5 +/- 0.6%) and testosterone-treated (4.2 +/- 0.8%) groups, but the mean changes did not differ between the groups. Linear regression analysis, however, demonstrated that the lower the pretreatment serum testosterone concentration, the greater the effect of testosterone treatment on lumbar spine bone density from 0-36 months (P = 0.02). This analysis showed a minimal effect (0.9 +/- 1.0%) of testosterone treatment on bone mineral density for a pretreatment serum testosterone concentration of 400 ng/dL (13.9 nmol/L), but an increase of 5.9 +/- 2.2% for a pretreatment testosterone concentration of 200 ng/dL (6.9 nmol/L). Increasing the serum testosterone concentrations of normal men over 65 yr of age to the midnormal range for young men did not increase lumbar spine bone density overall, but did increase it in those men with low pretreatment serum testosterone concentrations. PMID- 10372696 TI - Final height after long-term treatment with triptorelin slow release for central precocious puberty: importance of statural growth after interruption of treatment. French study group of Decapeptyl in Precocious Puberty. AB - The impact of treatment of central precocious puberty (CPP) with GnRH agonists on final statural height (FH) remains controversial, and guidelines on the optimal time point for interruption of these treatments have not been established. We analyzed the long term results of 58 girls and 8 boys uniformly treated with triptorelin slow release formulation (Decapeptyl, triptorelin-SR) for CPP and compared their FH with predicted height before treatment and with the FH of a historical group of patients not treated with GnRH agonist. The FH SD score was close to 0 and was not different from the genetic target height. In girls, FH was improved by 4.8 +/- 5.8 cm compared with predicted height before treatment and by 8.3 cm by comparison with a historical group. In boys, comparison with a historical group revealed a 13.7-cm improvement, whereas predicted height before treatment was similar to FH. Three variables were independently associated with FH in girls: the bone age/statural age ratio at the onset of treatment (negatively), the height SD score at the end of treatment, and the posttreatment growth spurt (delta FH - height at the end of treatment). The influence of the posttreatment growth spurt, itself dependent on age and bone age at the interruption of treatment, suggests that continuing treatment beyond the age of 11 yr in girls does not improve and could actually decrease FH. This point should be evaluated in a formal controlled trial. PMID- 10372697 TI - Effects of sleep and sleep deprivation on catecholamine and interleukin-2 levels in humans: clinical implications. AB - The objective of this study was to evaluate the effects of nocturnal sleep, partial night sleep deprivation, and sleep stages on catecholamine and interleukin-2 (IL-2) levels in humans. Circulating levels of catecholamines and IL-2 were sampled every 30 min during 2 nights: undisturbed, baseline sleep and partial sleep deprivation-late night (PSD-L; awake from 0300-0600 h) in 17 healthy male volunteers. Sleep was monitored somnopolygraphically. Sleep onset was associated with a significant (P < 0.05) decline of circulating concentrations of norepinephrine and epinephrine, with a nocturnal nadir that occurred 1 h after nocturnal sleep. On the PSD-L night, levels of norepinephrine and epinephrine significantly (P < 0.05) increased in association with nocturnal awakening. During stage 3-4 sleep, levels of norepinephrine, but not epinephrine, were significantly lower (P < 0.05) compared to average levels during the awake period, stages 1-2 sleep, and rapid eye movement sleep. Nocturnal levels of circulating IL-2 did not change with sleep onset or in relation to PSD-L or the various sleep stages. We conclude that sleep onset is associated with changes in levels of circulating catecholamines. Loss of sleep and disordered sleep with decreases in slow wave sleep may serve to elevate nocturnal catecholamine levels and contribute to cardiovascular disease. PMID- 10372698 TI - Effect of growth hormone (GH) and insulin-like growth factor I on prostate diseases: an ultrasonographic and endocrine study in acromegaly, GH deficiency, and healthy subjects. AB - The role of insulin-like growth factor I (IGF-I) in prostate development is currently under thorough investigation because it has been claimed that IGF-I is a positive predictor of prostate cancer. To assess the effect of GH and IGF-I levels on prostate pathophysiology, 46 acromegalic (30 in active disease, 10 cured from acromegaly, and 6 affected from GH deficiency) and 30 age-matched male controls, free from previous or concomitant prostate disorders, underwent pituitary, androgen, and prostate hormonal assessments and transrectal ultrasonography. Compared to control values, GH (P < 0.0001), IGF-I (P < 0.0001), and IGFBP-3 (P < 0.001) levels were increased, whereas testosterone (P < 0.0001) and dihydrotestosterone levels (P < 0.0001) were reduced in active acromegalic patients. Hypogonadism was present in 28 of the 46 acromegalic patients (60.8%). The anteroposterior (P < 0.05), and transverse (P < 0.0001) prostate diameters and the transitional zone volume (P < 0.05) were increased in acromegalic patients compared to those in controls. Prostate volume (PV) was significantly higher in untreated acromegalic patients than in controls (41.7 +/- 3.2 vs. 21.9 +/- 1.4 mL; P < 0.0001), cured patients (23.6 +/- 1.6 mL; P < 0.0001), and GH deficient patients (17.5 +/- 1.1 mL; P < 0.0001). In the patients, PV was correlated with disease duration (r = 0.606; P < 0.0001) and age (r = 0.496; P < 0.0001), whereas in controls it was correlated with age (r = 0.476; P < 0.01) and IGF-I levels (r = -0.448; P < 0.05). Benign prostate hyperplasia (PV > or = 30 mL) was found in 58% of the acromegalics and 26.6% of the controls. When grouped by age (<40, 40-60, and >60 yr), PV was increased in elderly patients compared to younger patients (P < 0.05) and to controls (P < 0.01). The prevalence of structural abnormalities, including calcifications, nodules, cysts, and vesicle inflammation, was significantly increased in patients compared to controls (78.2% vs. 23.3%; chi2 = 5.856; P < 0.05). No clinical, transrectal ultrasonography, or cytological evidence of prostate cancer was detected in acromegalic or control subjects. In conclusion, chronic excess of GH and IGF-I cause prostate overgrowth and further phenomena of rearrangement, but not prostate cancer. PMID- 10372699 TI - Prevention of bone demineralization by calcium supplementation in precocious puberty during gonadotropin-releasing hormone agonist treatment. AB - We have previously demonstrated a negative impact on peak bone mass in girls with precocious puberty treated with GnRH agonist (GnRHa). Several studies have shown that a high calcium intake positively influences bone mass in prepubertal girls and leads to a higher peak bone mass. The aim of this study was to evaluate the effect of calcium supplementation in girls with precocious puberty during GnRHa treatment. Forty girls affected by true central precocious puberty and treated with the GnRHa triptorelin were studied for 2 yr. After diagnosis, the patients were randomly assigned to three groups: group A, treated only with GnRHa; group B, treated for 12 months solely with GnRHa and then supplemented with calcium gluconolactate/carbonate (1 g calcium/day in two doses) for 12 months; and group C, treated from the beginning with combined GnRHa and calcium. Bone mineral density (BMD) at the lumbar spine was measured by dual energy x-ray absorptiometry at the beginning of the study and after 12 and 24 months and was expressed as the calculated true volumetric density (BMDv) in milligrams per cm3. Group A showed a decrease in absolute BMDv levels, in SD score for chronological age (CA), and even more in SD score for bone age (BA). Group B showed the same behavior during the first year, but this trend was reversed in the second year, when calcium supplementation was added to GnRHa treatment. Group C showed an increase in absolute BMDv levels and in SD score for CA and BA. BMDv variations (expressed as absolute values, SD score for CA, and SD score for BA) became statistically significant at 24 months between groups C and A (P = 0.036, P = 0.032, and P = 0.025, respectively). The behavior of the lumbar spine BMDv in the three groups is consistent with a positive effect of calcium supplementation during GnRHa treatment. In calcium-supplemented patients, the normal process of bone mass accretion at puberty is preserved despite GnRHa treatment. Therefore, the reduction in BMD during GnRHa treatment in girls with precocious puberty is at least completely reversible and preventable if calcium supplementation is associated from the beginning. PMID- 10372700 TI - Hormone replacement therapy causes a respiratory alkalosis in normal postmenopausal women. AB - Menopause is associated with an increase in venous bicarbonate concentrations that is reversible with hormone replacement therapy (HRT). However, the mechanism underlying this effect is not known. To address this question, we studied the changes in acid-base indexes in the arterialized venous blood of normal postmenopausal women commencing conjugated equine estrogen (0.625 mg/day), medroxyprogesterone acetate (MPA; 5 mg/day), their combination, or placebo, in a double blind randomized controlled study over 3 months. Serum bicarbonate concentrations decreased significantly in the groups receiving either MPA or estrogen plus MPA (P = 0.008). This trend was apparent as early as 2 days and reached 2.7 and 2.3 mmol/L in the respective groups by 3 months. Similar changes were seen with partial pressure of carbon dioxide (P = 0.04); a change of -0.7 kPa occurred in the estrogen plus MPA group at 3 months. There were no changes in bicarbonate concentrations or partial pressure of carbon dioxide in those receiving estrogen alone or placebo. Accompanying changes in blood pH were apparent in the estrogen plus MPA group, where there was an upward trend at 1 week (P = 0.056) and a significant change from baseline (+0.013) at 3 months (P = 0.03). In the whole group, the changes in pH were inversely correlated with those in urinary excretion of hydroxyproline (r = -0.44; P = 0.01). We conclude that HRT using conjugated estrogens and MPA produces small, but sustained, changes in acid-base status. These may contribute to the effects of HRT and menopause on many tissues and disease processes, including the development of osteoporosis. PMID- 10372701 TI - Gender differences in the effects of long term growth hormone (GH) treatment on bone in adults with GH deficiency. AB - We recently observed that among patients with GH deficiency due to adult-onset hypopituitarism, men responded with a greater increase in serum levels of insulin like growth factor I (IGF-I) and biochemical markers of bone metabolism than women when the same dose of recombinant human GH (rhGH) per body surface area was administered for 9 months. In the present study, 33 of the 36 patients in the previous trial (20 men and 13 women) continued therapy for up to 45 months. The dose of rhGH was adjusted according to side-effects and to maintain serum IGF-I within the physiological range. This resulted in a significant dose reduction in the men; consequently, the women received twice as much rhGH as the men (mean +/- SD, 1.9 +/- 1.1 vs. 1.0 +/- 0.6 U/day; P < 0.01). The increases in serum IGF-I levels and serum biochemical markers of bone metabolism were similar in men and women with these doses. The total bone mineral content (BMC) was increased after 33 and 45 months of treatment up to 5.1% (P = 0.004 and 0.0001). Bone mineral density (BMD), BMC, and the area of the femoral neck and the lumbar spine were also significantly increased after 33 and 45 months of treatment. When analyzed by gender, total body BMC, femoral neck BMD and BMC, and spinal BMC were significantly increased in males, but not in females (P < 0.05-0.01). In conclusion, rhGH treatment continued to have an effect on bone metabolism and bone mass for up to 45 months of therapy. The changes in bone mass were greater in the men, although they received lower doses of rhGH than the women. The results indicate that the sensitivity to GH in adult patients with GH deficiency is gender dependent. PMID- 10372702 TI - Endogenous androgens and carotid intimal-medial thickness in women. AB - The influence of endogenous androgens on atherosclerotic disease in women is unknown. In this study involving 101 pre- and post-menopausal females, we evaluated the relationship between serum androgen levels and both carotid artery intimal-medial thickness (IMT) and major cardiovascular risk factors. In addition to evaluation of blood pressure, body mass index, and waist-to-hip ratio, serum dehydroepiandrosterone sulfate (DHEA-S), androstenedione (A), total testosterone (TTS), free testosterone (FTS), insulin, cholesterol (total and high density lipoproteins), triglycerides, and glucose were measured. All women underwent carotid ultrasonography. Spearman correlation coefficients showed that serum DHEA S and A levels were negatively related (P < 0.03-0.0004) to several IMT measures. Higher tertiles of DHEA-S, A, and FTS corresponded to significantly lower measures of carotid thickness. DHEA-S, and all androgens were inversely related to age (P < 0.03 or less), showing no unfavorable association with major cardiovascular risk factors. In contrast, serum DHEA-S was negatively associated with WHR (P < 0.02), while A was negatively associated with body mass index (P < 0.02). Stepwise multiple regression analysis indicated that A and FTS showed an inverse association with IMT measures (P < 0.05-0.001). In conclusion, our data indicate that in women serum DHEA-S and androgens decline with age and that normal hormonal levels are not associated with major cardiovascular risk factors. They also show that higher DHEA-S and androgen concentrations are related to lower carotid wall thickness; for A this association is independent of cardiovascular risk factors. Our results suggest that, in the physiological range, DHEA-S and androgens in women are correlated with lower risk of carotid artery atherosclerosis. PMID- 10372703 TI - Serum levels of insulin-like growth factor I in 152 patients with growth hormone deficiency, aged 19-82 years, in relation to those in healthy subjects. AB - Serum insulin-like growth factor I (IGF-I) levels within normal range for age have been reported to be common in adults with GH deficiency (GHD). Therefore, serum IGF-I levels were determined in 152 consecutive patients (71 women and 81 men) with evidence of hypothalamic-pituitary disorders or previous cranial radiation, who fulfilled the presently used criteria for GHD i.e. peak GH response below 3 microg/L at stimulation test. Patients treated for acromegaly were excluded. Forty-three patients, aged 19-63 yr, had childhood onset GHD, and 109, aged 23-82 yr, had adult-onset GHD. Their IGF-I levels were expressed in SD scores in relation to normal reference values based on 448 healthy subjects, aged 20-96 yr (247 women and 201 men). In healthy subjects a linear inverse correlation, without gender difference, was found between logarithmic transformed IGF-I levels and age (r = -0.774; P < 0.001). In contrast, no age dependency was found in GHD patients. All patients with childhood-onset GHD had IGF-I values below -2 SD, significantly lower than those in adult-onset GHD patients (-6.2 +/- 0.3 vs. -3.2 +/- 0.2 SD score; P < 0.001). In patients with adult-onset GHD, 34% of the IGF-I levels were within normal range, increasing to 40% in the subgroup above 60 yr of age, in whom 86% were diagnosed with hypothalamic-pituitary tumors. Normal IGF-I was more common in men than in women, but no difference was observed between patients with panhypopituitarism and those with partial pituitary insufficiency. High frequencies of IGF-I levels within the normal range were found in GHD patients with pituitary tumors (20 of 57 nonsecreting pituitary adenomas, 5 of 15 prolactinomas, 6 of 12 Cushing's disease, and 4 of 25 craniopharyngiomas), but in only 2 of 43 patients with GHD due to other causes. In conclusion, an IGF-I level below -2 SD seems to be of diagnostic value in GHD with onset in childhood or early adulthood, whereas values within normal range are common in patients over 60 yr of age, especially those with pituitary tumors. The outcome of GH replacement therapy may reveal whether the addition of IGF-I as a diagnostic criterion is of predictive value in older patients. PMID- 10372704 TI - Estrogen activity and novel tissue selectivity of delta8,9-dehydroestrone sulfate in postmenopausal women. AB - Recent basic and clinical advances have consolidated the concept of tissue selective estrogens, i.e. molecules that express different degrees of partial agonist, full agonist or antagonist activity in different tissues or cells. Delta8,9-Dehydroestrone sulfate (delta8,9-DHES) is a conjugated estrogen and a component of conjugated equine estrogens (CEE). It is metabolized in the human in at least a 1:1 ratio to its 17beta form, 17beta-delta8,9-DHES. To evaluate its activity in different clinical and biochemical parameters, a clinical research study was conducted with delta8,9-DHES and estrone sulfate as a comparator in postmenopausal women. Delta8,9-DHES was given orally at a daily dose of 0.125 mg for 12 weeks in a group of 10 women. Two additional groups of women received either estrone sulfate alone (1.25 mg/day) or the combination of delta8,9-DHES and estrone sulfate at the previously specified doses. A significant and consistent suppression of hot flushes (number, severity, and total score) was observed with delta8,9-DHES, reaching more than 95% suppression in all parameters of vasomotor symptoms. This level of activity was equal to that obtained with the much higher dose of estrone sulfate, and it was sustained for the duration of the treatment period (12 weeks). Measurements of a bone resorption marker, i.e. urinary excretion of N-telopeptide, demonstrated that delta8,9-DHES at 8 weeks produced a degree of suppression (40%) similar to that observed with the higher dose of estrone sulfate. Gonadotropin secretion (FSH and LH) was significantly suppressed in women receiving delta8,9-DHES, similar to that observed with estrone sulfate alone or with the combination of the two. Other parameters, such as total cholesterol, low density lipoprotein cholesterol and high density lipoprotein cholesterol were not modified significantly, whereas serum globulins (sex hormone-binding globulin and corticosteroid-binding globulin) showed only marginal increases after delta8,9-DHES administration. Taken together with preclinical data, it is found that delta8,9-DHES is an active estrogen with a distinct pharmacological profile that results in significant clinical activity in vasomotor, neuroendocrine (gonadotropin and PRL) and bone preservation parameters, whereas displaying little or no efficacy, at the dose tested, on other peripheral parameters normally affected by estrogens. Collectively, this information supports the concept that delta8,9-DHES is an integral component of CEE, with distinct tissue selectivity contributing to the CEE's overall clinical activity, and places this estrogen as a distinct member of a novel class of centrally active molecules with unique peripheral tissue selectivity. PMID- 10372705 TI - Treatment of obese subjects with the oral growth hormone secretagogue MK-677 affects serum concentrations of several lipoproteins, but not lipoprotein(a). AB - Obesity is associated with blunted GH secretion and an unfavorable lipoprotein pattern. The objective of this study was to investigate the effects of treatment with the oral GH secretagogue MK-677 on lipoproteins in otherwise healthy obese males. The study was randomized, double blind, and parallel. Twenty-four obese males, aged 18-50 yr, with body mass index greater than 30 kg/m2 and waist/hip ratio above 0.95 were treated with 25 mg MK-677 (n = 12) or placebo (n = 12) daily for 8 weeks. MK-677 treatment did not significantly change serum lipoprotein(a) [Lp(a)] levels. Serum apolipoprotein A-I and E (apoA-I and apoE) were increased at 2 weeks (P < 0.001 and P < 0.01 vs. placebo, respectively), but were not changed at study end. Serum total cholesterol and low density lipoprotein (LDL) cholesterol (LDL-C) levels were not significantly changed by MK 677 treatment. Serum high density lipoprotein (HDL) cholesterol (HDL-C) was increased at 2 weeks of MK-677 treatment (P < 0.01 vs. placebo), but not at 8 weeks. The LDL-C/HDL-C ratio was reduced after 8 weeks of MK-677 treatment (P < 0.05 vs. placebo). Mean LDL particle diameter was decreased at 2 weeks (P < 0.05 vs. placebo), but was unchanged compared with baseline values at 8 weeks (P = NS vs. placebo). The level of serum triglycerides was increased at 2 (P < 0.05 vs. placebo), but not at 8, weeks. Lipoprotein lipase activity in abdominal and gluteal sc adipose tissue was not affected by active treatment. In conclusion, treatment with the oral GH secretagogue MK-677 affected circulating lipoproteins. The effects on serum apoA-1, apoE, triglycerides, and mean LDL particle diameter were transient. At study end, the LDL-C/HDL-C ratio was decreased. MK-677 treatment did not significantly affect serum Lp(a) concentrations at the present dose and administration protocol. PMID- 10372706 TI - Decreased serum levels of acid-labile subunit in patients with anorexia nervosa. AB - One of the observations in malnutrition is that serum insulin-like growth factor (IGF)-I levels are decreased, and this decrease is associated with an altered profile of IGF binding proteins (IGFBPs). In human circulation, IGFs are mostly present as an approximately 150-kDa ternary protein complex consisting of IGFs, IGFBP-3, and acid-labile subunit (ALS). In the present study, to clarify the effect of nutrition on serum ALS levels, we investigated 33 patients with anorexia nervosa. Serum levels of ALS were measured by RIA. Furthermore, we measured serum IGF-I, IGF-II, IGFBP-2, and IGFBP-3 levels in the patients. From these data, we investigated which was the best predictor of body mass index (BMI) as a nutritional status marker. In the patients with anorexia nervosa, the serum ALS levels ranged from 0.7-16.9, with a mean of 10.6 +/- 0.7 mg/L, and the levels were significantly lower than those of normal subjects (13.8 +/- 0.8 mg/L, P < 0.05). Serum ALS levels positively correlated with BMI (r = 0.41, P < 0.05), and the levels increased during treatment. The serum IGFBP-2 levels in the patients were increased (871 +/- 91 microg/L), and the levels inversely correlated with BMI (r = -0.52, P < 0.01). The serum IGF-I and IGFBP-3 levels were low (152 +/- 14 microg/L and 2.56 +/- 0.12 mg/L, respectively), and the levels positively correlated with BMI (r = 0.46, P < 0.01; and r = 0.39, P < 0.05, respectively). The serum IGFBP-2, IGF-I, and IGFBP-3 levels returned toward normal ranges as BMI in the patients improved during treatment. Serum IGF-II levels did not correlate with BMI (r = 0.24, P = 0.17). Stepwise regression analysis revealed that serum IGFBP-2 was the best marker of BMI among these variables. The present study suggested that ALS was regulated by nutritional status, the same as IGF-I, IGFBP 2 and IGFBP-3; but the serum IGFBP-2 was the best predictor of BMI as nutritional status marker among the parameters in patients with anorexia nervosa. PMID- 10372707 TI - The roles of insulin sensitivity, insulin-like growth factor I (IGF-I), and IGF binding protein-1 and -3 in the hyperandrogenism of African-American and Caribbean Hispanic girls with premature adrenarche. AB - Recent reports indicate that girls with premature adrenarche are at risk of developing functional ovarian hyperandrogenism and polycystic ovarian syndrome (PCOS). As insulin and insulin-like growth factors (IGFs) have been implicated in the pathogenesis of PCOS, we hypothesize that they may also have a role in the hyperandrogenism of premature adrenarche. Thirty-five prepubertal girls (23 Caribbean Hispanics and 12 Black African-Americans) underwent a 60-min ACTH and LH-releasing hormone test. Insulin sensitivity (S(I)) was assessed using the frequently sampled i.v. glucose tolerance test with tolbutamide. Fasting levels of IGF-I, IGF-binding protein-1 (IGFBP-1), IGFBP-3, sex hormone-binding globulin, and free testosterone (T) were also obtained. The mean age of the patients was 6.8 yr, and bone age was 8.0 yr. Twenty-five patients had a family history of noninsulin-dependent diabetes mellitus and 19 patients had acanthosis nigricans. The mean S(I) for the entire group was 6.78 +/- 5.21 x 10(-4) min/microU x mL (normal prepubertal S(I), 6.5 +/- 0.54 x 10(-4) min(-1) x microU(-1) x mL(-1)). However, 15 of the 35 girls had an S(I) that was more than 2 SD below the mean reported for normal prepubertal children. Of these 15 patients, 13 were obese, and 14 had acanthosis nigricans. For the entire group of girls, the mean ACTH stimulated levels of 17-hydroxypregnenolone (17OHPreg), dehydroepiandrosterone (DHEA), androstenedione (AS), 17-hydroxyprogesterone (17OHP), and T and the ACTH stimulated ratios of 17OHPreg/17OHP, 17OHPreg/DHEA, 17OHP/AS, and DHEA/AS did not differ from the levels reported for Tanner stage II-III pubertal girls. The girls were divided into two groups based on their S(I) (group I, S(I) >2 SD below the mean for age; group II, normal S(I)). The group I girls with a reduced S(I) had significantly higher ACTH-stimulated levels of 17OHPreg (group I, 760 +/- 87.84 ng/dL; group II, 428.9 +/- 46.28 ng/dL; P = 0.002), 17OHPreg/17OHP ratio (group I, 3.95 +/- 0.36; group II, 2.96 +/- 0.35; P = 0.05), 17OHPreg/DHEA (group I, 2.06 +/- 0.21; group II, 1.4 +/- 0.13; P = 0.01), and free T (group I, 1 +/- 0.23 ng/dL; group II, 0.49 +/- 0.19 ng/dL; P = 0.014). Levels of sex hormone-binding globulin were lower in the group I girls. Furthermore, for the entire group of girls, the S(I) correlated inversely with ACTH-stimulated levels of 17OHPreg, DHEA, and AS and the ACTH-stimulated ratio of 17OHPreg/17OHP. IGF-I correlated inversely with S(I) (r = -0.94; P < 0.001) and correlated directly with the ACTH stimulated levels of 17OHPreg (r = 0.8; P < 0.001) and AS (r = 0.63; P < 0.05). IGF-I also correlated with the ACTH-stimulated ratios of 17OHPreg/17OHP (r = 0.61; P < 0.05), 17OHPreg/DHEA (r = 0.9; P < 0.001), 17OHP/AS (r = 0.79; P < 0.001), and DHEA/AS (r = 0.96; P < 0.001). IGFBP-1 correlated inversely with the ACTH-stimulated levels of 17OHPreg (r = -0.38; P < 0.05) and DHEA (r = -0.36; P < 0.05). To summarize, the ACTH-stimulated delta5-steroid levels were higher in prepubertal girls with premature adrenarche and reduced S(I). There was a significant inverse correlation among ACTH-stimulated hormone levels, S(I), and IGFBP-1, whereas IGF-I correlated directly with ACTH-stimulated androgens. These findings support the hypothesis that insulin and IGFs may have a role in the hyperandrogenism of premature adrenarche just as they do in PCOS. Hence, in certain girls with premature adrenarche, hyperandrogenism may be the first presentation of PCOS and/or insulin resistance. PMID- 10372708 TI - Bone mineral mass and calcium and phosphate metabolism in young men: relationships with vitamin D receptor allelic polymorphisms. AB - The genetic bases of peak bone mass determinants are still poorly understood, particularly in males. We investigated the relationship between vitamin D receptor (VDR) 3'- and 5'-gene polymorphisms (as determined by the restriction enzymes BsmI and FokI) and bone mineral mass, and calcium and inorganic phosphate (Pi) metabolism in an homogeneous cohort of young healthy men. In 104 healthy subjects, aged 24.3 +/- 3.1 yr (mean +/- SD; range, 20.7-38.7), standardized bone mineral density (BMD; z-scores) at the levels of lumbar spine and femoral trochanter, i.e. bone mineral content adjusted for age, body size, and bone area, significantly differed between VDR 3'-end alleles (BsmI), whereas crude areal BMD or bone mineral content did not. Among BsmI homozygotes BB, BMD (z-scores) were significantly lower only in subjects also carrying the f allele at the VDR-5' polymorphic site (FokI). Serum PTH levels were significantly higher in the BB genotype at baseline and remained so under either a low or a high calcium phosphorus diet. Moreover, on the low calcium-phosphorus diet, BB subjects had significantly decreased tubular Pi reabsorptive capacity and plasma Pi levels. Our results underline the importance of identifying multiple single base mutation polymorphisms within candidate genes of bone mineral mass and suggest a role for environmental/dietary factor interactions with VDR gene polymorphisms in peak bone mineral mass in men. PMID- 10372709 TI - Severity of osteopenia in estrogen-deficient women with anorexia nervosa and hypothalamic amenorrhea. AB - Reduced bone density is observed in over half of women with anorexia nervosa (AN), in whom the risk of fracture is significantly increased even at a young age. It is unknown to what extent low bone density in AN differs from other conditions of premenopausal osteoporosis and is related to estrogen deficiency and/or other factors, such as nutritional status. We therefore investigated bone loss in nutritionally replete and nutritionally deplete amenorrheic women by comparing patients with AN (n = 30) to age-matched subjects with hypothalamic amenorrhea (HA; n = 19) in whom duration of amenorrhea, prior estrogen use, and age of menarche were comparable. Healthy, age-matched, eumenorrheic women were studied as a control group (NL; n = 30). Weight and nutritionally dependent factors including (body mass index, 20.7 +/- 0.3 vs. 16.7 +/- 0.3 kg/m2; P < 0.0001), insulin-like growth factor I (270 +/- 18 vs. 203 +/- 17 ng/mL; P < 0.01), percent body fat (26% vs. 19%; P < 0.0001), and lean body mass (38.7 +/- 1.1 vs. 34.3 +/- 0.8, P < 0.01) were significantly different between the HA and AN groups, respectively. The bone densities of the anterior-posterior (AP) spine, total hip, and total body measured by dual energy x-ray absortiometry were reduced in both amenorrheic groups compared to those in control subjects, but were significantly lower in women with AN than in those with HA. The t scores for AP spine and hip were -1.80 +/- 0.15 (AN), -0.80 +/- 0.22 (HA), and 0.28 +/- 0.19 SD (NL) for the AP spine and -1.62 +/- 0.17 (AN), -0.51 +/- 0.21 (HA), and 0.25 +/- 0.16 (NL) for the total hip, respectively (P < 0.01 for all comparisons). Among the amenorrheic subjects, duration of amenorrhea, age of menarche, and N telopeptide were inversely correlated with bone density at all sites, whereas body mass index, insulin-like growth factor I, lean body mass, and fat intake were positively correlated with bone density at all sites measured. In multivariate regression analyses, bone density was most significantly related to lean body mass (P = 0.05 and P = 0.03 for the spine and hip, respectively), but not to the duration of amenorrhea or other indexes of estrogen status among patients with AN. In contrast, bone density of the lumbar spine was significantly related to weight and duration of amenorrhea among patients with HA. These data demonstrate that the severity of osteopenia in AN is greater than that in patients with HA and is critically dependent upon nutritional factors in addition to the degree or duration of estrogen deficiency itself. Lean body mass, independent of the duration or severity of estrogen deficiency, is an important predictor of bone loss among women with AN. PMID- 10372710 TI - Jointly amplified basal and pulsatile growth hormone (GH) secretion and increased process irregularity in women with anorexia nervosa: indirect evidence for disruption of feedback regulation within the GH-insulin-like growth factor I axis. AB - Anorexia nervosa (AN) is associated with multiple endocrine alterations. In the majority of AN patients, basal and GHRH-stimulated serum GH levels are increased. The metabolic effects of GH are known to be related to its pulsatile secretory pattern. The present study was performed to examine GH pulsatility in AN using the techniques of deconvolution analysis and approximate entropy, which quantify secretory activity and serial irregularity of underlying hormone release not reflected in peak occurrence or amplitudes. To this end, 24-h GH profiles were obtained by continuous blood sampling aliquoted at 20-min intervals in 8 nonfasting patients with AN [body mass index (BMI), 14.2 +/- 0.8 kg/m2; mean +/- SEM) and in 11 age-matched healthy women (BMI, 20.3 +/- 0.5 kg/m2). The deconvolution-estimated half-life of GH was not altered in the AN patients. The pituitary GH secretory burst frequency, burst mass, and burst duration were each significantly increased in women with AN compared to those in normal weight women. A 4-fold increase in daily pulsatile GH secretion was accompanied by a 20 fold increase in basal (nonpulsatile) GH secretion. There were significant negative correlations between BMI and the basal as well as pulsatile GH secretion rates. Moreover, AN patients exhibited significantly greater GH approximate entropy scores than the controls, denoting marked irregularity of the GH release process. In contrast to previous reports in healthy fasting subjects, cortisol levels in AN patients were positively correlated to GH secretion rates. Leptin levels were significantly inversely correlated to the pulsatile, but not the basal, GH secretion rate. The present data demonstrate augmented basal as well as pulsatile GH secretion with disruption of the orderliness of the GH release process in AN. Accordingly, GH secretion in AN probably reflects altered neuroendocrine feedback regulation, e.g. associated with increased hypothalamic GHRH discharge superimposed on reduced hypothalamic somatostatinergic tone. PMID- 10372711 TI - Left ventricular diastolic dysfunction in patients with subclinical hypothyroidism. AB - Although subclinical hypothyroidism is frequently diagnosed, the decision to institute a substitutive therapy with L-T4 remains controversial. Because the cardiovascular system is considered a main target for the action of thyroid hormone, we investigated whether subclinical hypothyroidism induces cardiovascular abnormalities. Twenty-six patients (mean age, 36 +/- 12 yr) were evaluated by Doppler-echocardiography, whereas a subgroup of 10 patients, randomly selected, were reevaluated after 6 months of L-T4 substitutive therapy (mean dose, 68 microg daily). Thirty subjects (matched for age, sex, and body surface area) served as controls. Mean plasma TSH was significantly higher in patients (P < 0.001), whereas mean serum free T4 and free T3 concentrations, although in the normal range, were significantly lower (P < 0.001 and P < 0.005, respectively). Blood pressure and heart rate did not differ from control values. Echocardiogram examination showed no abnormalities of the left ventricular morphology and a slight, but not significant, reduction in the systolic function in the patient group. In contrast, Doppler-derived indices of diastolic function showed significant prolongation of the isovolumic relaxation time (94 +/- 13 vs. 84 +/- 8 msec; P < 0.001), increased A wave (55 +/- 13 vs. 48 +/- 9 cm/sec; P < 0.05), and reduced early diastolic mitral flow velocity/late diastolic mitral flow velocity ratio (1.4 +/- 0.3 vs. 1.7 +/- 0.3; P < 0.001). In the subgroup of 10 patients, thyroid hormone profile was normalized by 6 months of L-T4 substitutive therapy, whereas no changes were observed in the left ventricular morphology. Systolic function was significantly enhanced, as compared with pretreatment values (P < 0.01) but did not differ from control values. Also, systemic vascular resistance was significantly decreased by L-T4 replacement therapy. Assessment of diastolic function showed significant shortening of isovolumic relaxation time (77 +/- 15 vs. 91 +/- 8; P < 0.05), reduction of A wave (51 +/- 13 vs. 60 +/- 12; P < 0.01), and increase of early diastolic mitral flow velocity/late diastolic mitral flow velocity ratio (1.7 +/- 0.4 vs. 1.3 +/- 0.3; P < 0.001). These indices, however, were comparable with those of control subjects. These findings indicate that subclinical hypothyroidism affects diastolic function and that this abnormality may be reversed by L-T4 substitutive therapy. PMID- 10372712 TI - Relative hyperproinsulinemia as a sign of islet dysfunction in women with impaired glucose tolerance. AB - Proinsulin release is increased relative to insulin secretion in subjects with type 2 diabetes, indicative of islet dysfunction. However, it has not been conclusively shown whether there is an increased relative proinsulin release in subjects with impaired glucose tolerance (IGT), i.e. whether it precedes the development of diabetes. We therefore determined the proinsulin to insulin ratios in the fasting state and after acute stimulation of insulin secretion in 23 postmenopausal women, aged 61-62 yr (mean +/- SD, 61.7 +/- 0.5 yr). Ten women had normal glucose tolerance (NGT), and 13 had IGT. The groups were matched for insulin sensitivity and did not differ in body mass index. Proinsulin and insulin secretion were measured after arginine stimulation (5 g, i.v.) at three glucose levels (fasting, 14 mmol/L, and >25 mmol/L), and the acute insulin (AIR(arg)) and proinsulin responses (APIR(arg)) were calculated as the mean 2-5 min postload increase. At fasting glucose, levels of insulin, proinsulin, or the proinsulin/insulin ratio (13.6 +/- 5.0% vs. 11.1 +/- 2.7%; P = NS) did not differ between NGT and IGT. Although the AIR(arg) values were decreased in the IGT group at all glucose levels (P < 0.05), the absolute proinsulin levels and the APIRs(arg) were similar between IGT and NGT women. Therefore, the IGT women had higher proinsulin/insulin ratios at 14 mmol/L (10.7 +/- 4.4% vs. 6.4 +/- 1.8%; P = 0.006) and more than 25 mmol/L glucose (11.4 +/- 5.2% vs. 6.7 +/- 2.1%; P = 0.007). The IGT group had increased APIR(arg)/AIR(arg) at fasting (2.2 +/- 1.4% vs. 1.3 +/- 0.6%; P = 0.047) and more than 25 mmol/L glucose (3.5 +/- 1.6% vs. 2.3 +/- 0.7%; P = 0.037). We conclude that women with IGT exhibit increased relative proinsulin secretion, suggesting a defect in the intracellular proinsulin processing before diabetes develops. PMID- 10372713 TI - Leptin secretion in Cushing's syndrome: preservation of diurnal rhythm and absent response to corticotropin-releasing hormone. AB - The normal inverse relationship between leptin and cortisol is lost in chronic hypercortisolism. We studied this apparent dysregulation in patients with Cushing's syndrome to investigate 1) the effect of chronic hypercortisolemia on the circadian rhythm of leptin secretion, 2) the response of leptin after administration of CRH, and 3) the short term effect of curative surgery on leptin. The preoperative morning leptin concentration was 54.2 +/- 8.1 ng/mL, and the nighttime value was 68.6 +/- 9.8 ng/mL, reflecting a mean rise of 32.8 +/- 7.6%, similar to the nocturnal increase observed in normal subjects. By contrast, cortisol's diurnal variation (21.8 +/- 1.7 vs. 16.9 +/- 1.1 mg/dL) was blunted. In women, but not men, body mass index correlated with leptin (P = 0.001). Preoperative ACTH and cortisol (both P < 0.0001), but not leptin levels increased after CRH. Ten days after surgery, basal cortisol values were subnormal (1.1 +/- 0.6 mg/dL), but leptin levels remained unchanged and did not increase after CRH. Body mass index and insulin also remained unchanged. Insulin, but not age, urinary free cortisol, or plasma cortisol correlated with leptin (P < 0.05). In summary, patients with Cushing's syndrome have moderately elevated leptin levels that maintain an intact circadian rhythm but do not respond to acute or subacute alterations of cortisol. PMID- 10372715 TI - Laminin receptors in differentiated thyroid tumors: restricted expression of the 67-kilodalton laminin receptor in follicular carcinoma cells. AB - The expression of integrin laminin receptors was investigated in normal thyroid primary cultures; immortalized normal thyroid cells (TAD-2); papillary (NPA), follicular (WRO), and anaplastic (ARO) thyroid tumor cell lines; seven thyroid tumors (four papillary and three follicular carcinomas); and normal thyroid glands. The expression of alpha1beta1, alpha2beta1, alpha3beta1, alpha6beta1, and alpha6beta4 was found in all tumor specimens and in tumor cell lines, whereas normal thyroid cells and TAD-2 cells lacked the expression of alpha6beta4. Despite the presence of several integrin laminin receptors, adhesion of TAD-2, NPA, and ARO cells to immobilized laminin-1 was poor, whereas WRO cells and follicular carcinoma-derived cells displayed a strong adhesion. Indeed, WRO and follicular carcinoma-derived cells showed expression of a nonintegrin laminin receptor, the 67-kDa high affinity laminin receptor (67LR). TAD-2, NPA, and ARO cells as well as nodular goiter, toxic adenoma, follicular adenoma, and papillary carcinoma-derived cells did not express the 67LR. Adhesion of WRO and follicular carcinoma-derived cells to laminin-1 was specifically inhibited by a recombinant polypeptide containing laminin-binding domains of 67LR, demonstrating that this receptor confers to follicular carcinoma cells attachment capacity to laminin. Moreover, tissue specimens from follicular carcinomas expressed the 67LR, whereas follicular adenomas and normal thyroid tissues were negative. In thyroid tumors, integrin receptors, although abundant, participate weakly in adhesion to laminin. The expression in follicular carcinoma cells of a functional, high affinity 67LR together with nonfunctional integrin LM receptors could be responsible for the tendency of follicular carcinoma cells to metastasize by mediating stable contacts with basal membranes. PMID- 10372714 TI - Loss of estrogen inactivation in colonic cancer. AB - Age and sex differences in the incidence of colonic cancer, together with epidemiological data on patients taking hormone replacement therapy, suggest the involvement of estrogens. Analogous to the role of aromatase in breast cancer, we postulated that steroid metabolism within the colon itself may be a crucial mechanism in regulating tissue exposure to estrogens. We have characterized expression of aromatase (responsible for converting C19 androgens to C18 estrogens) and 17beta-hydroxysteroid dehydrogenase (17beta-HSD) [responsible for interconversion of active estradiol (E2) to less potent estrone (E1)] in normal and neoplastic human colon from 24 patients undergoing tumor resection. Aromatase activity was similar in homogenates from normal mucosa, tissue adjacent to tumors, and the tumors themselves. Analysis of 17beta-HSD activity indicated that the predominant activity was oxidative (E2 to E1), and this conversion was significantly lower in colonic tumors [444 (90-1735); median (95% confidence interval) pmol/mg protein x h], compared with normal mucosa [1709 (415-13828), P < 0.001]. Northern blot analyses indicated expression of messenger RNAs (mRNAs) for the type 2 and 4 isozymes of 17beta-HSD in normal colon; messenger RNA for 17beta-HSD 4 was significantly lower in tumor tissue [0.75 +/- 0.22 (mean +/- SD) arbitrary U vs. 0.43 +/- 0.17, P < 0.01]. Studies in vitro, using three colonic cancer cell lines, indicated that there was an inverse correlation between 17beta HSD oxidative activity and the rate of cell proliferation. In addition, E1, but not E2, was shown to significantly decrease proliferation when added exogenously to the colonic epithelial cell line, SW620 cells. Colonic mucosa can regulate estrogen hormone action in an intracrine fashion. The loss of estrogen inactivation may be an important mechanism in the pathogenesis of colonic cancer. PMID- 10372716 TI - Linkage and association of the sodium potassium-adenosine triphosphatase alpha2 and beta1 genes with respiratory quotient and resting metabolic rate in the Quebec Family Study. AB - The purpose of this study was to examine the relationship between the alpha2 (exon 1 and exon 21-22 with BglII) and beta1 (MspI and PvuII) genes of the sodium potassium adenosine triphosphatase and resting metabolic rate (RMR) and respiratory quotient (RQ). The sample included 582 participants from 171 families of the Quebec Family Study. RMR and RQ were adjusted for age, sex, fat mass, and fat free mass. Sib-pair analyses indicated a significant linkage between RQ and the alpha2 exon 1 marker (P = 0.03) and the alpha2 exon 21-22 marker (P = 0.02). No linkage was detected between the beta1 markers and either RMR or RQ, whereas RMR was not linked with the alpha2 makers. There was a significant interaction (P < 0.0003) between alpha2 exon 1 carrier status and age group [younger (< 45 yr) vs. older (> or = 45 yr) adults] for RQ. The association between carrier status and RQ was significant in younger adults (RQ = 0.76 in carriers vs. 0.80 in noncarriers, P < 0.0001) but was not in older adults (RQ = 0.81 in carriers vs. 0.80 in noncarriers). The alpha2 exon 1 gene accounted for approximately 9.1% and 0.3% of the variance in RQ in younger and older adults, respectively. The results suggest that the sodium potassium adenosine triphosphatase alpha2 gene may play a role in fuel oxidation, particularly in younger individuals. PMID- 10372717 TI - Growth hormone (GH) receptor blockade with a PEG-modified GH (B2036-PEG) lowers serum insulin-like growth factor-I but does not acutely stimulate serum GH. AB - B2036-PEG, a GH receptor (GH-R) antagonist, is an analog of GH that is PEG modified to prolong its action. Nine mutations alter the binding properties of this molecule, preventing GH-R dimerization and GH action. A potential therapeutic role of B2036-PEG is to block GH action, e.g. in refractory acromegaly. A phase I, placebo-controlled, single rising-dose study was performed in 36 normal young men (ages, 18-37 yr; within 15% ideal body weight). Four groups received a single s.c. injection of either placebo (n = 3 in each group, total n = 12) or B2036-PEG (0.03, 0.1, 0.3, or 1.0 mg/kg; n = 6 each dose). B2036 PEG and GH concentrations were measured 0, 0.25, 0.5, 1, 3, 6, 9, 12, 24, 36, 48, 72, 96, 120, and 144 h after dosing. Serum insulin-like growth factor-I was measured before and 1-7 days after dosing. All doses were well tolerated, with no serious or severe adverse reactions. B2036-PEG, at 1.0 mg/kg, reduced insulin like growth factor-I by 49 +/- 6% on day 5 (P < 0.001 vs. placebo). GH was measured by two independent methods: 1) modified Nichols chemiluminescence assay (empirically corrected for B2036-PEG cross-reactivity); and 2) direct GH two-site immunoassay, using monoclonal antibodies that did not react with B2036-PEG. There was good agreement between the two methods. GH did not change substantially at any B2036-PEG dose, suggesting that B2036-PEG does not interact with hypothalamic GH-Rs to block short-loop feedback. B2036-PEG may thus block peripheral GH action without enhancing its secretion. PMID- 10372719 TI - The effects of hormone replacement therapy on hypothalamic neuropeptide gene expression in a primate model of menopause. AB - Menopause is associated with increased neurokinin B (NKB) gene expression and decreased proopiomelanocortin (POMC) gene expression in the human hypothalamus. In the present study, young, ovariectomized cynomolgus monkeys were used in a model of menopause to examine the effects of hormone replacement therapy (HRT) on hypothalamic neuropeptide gene expression. A secondary goal was to determine whether HRT produces signs of estrogen toxicity in the primate hypothalamus by examining POMC neurons and microglial cells. In situ hybridization was performed using synthetic, radiolabeled, 48-base oligonucleotide probes. Alpha-napthyl butyrate esterase histochemistry was used to visualize microglial cells. Both estrogen and estrogen plus progesterone treatments produced a marked suppression of the number of infundibular neurons expressing NKB gene transcripts. In contrast, HRT had no effect on the POMC system of neurons or the number of microglial cells in the infundibular nucleus. These results provide strong support for the hypothesis that the increased NKB gene expression in the hypothalamus of postmenopausal women is secondary to estrogen withdrawal. Conversely, these data suggest that the dramatic decline in the numbers of neurons expressing POMC gene transcripts in older women is caused by factors other than ovarian failure. Finally, we found no evidence that HRT, in doses designed to mimic currently prescribed regimens, produces signs of estrogen toxicity in the primate infundibular nucleus. PMID- 10372718 TI - Medroxyprogesterone acetate and dexamethasone are competitive inhibitors of different human steroidogenic enzymes. AB - Medroxyprogesterone acetate (MPA), a widely used progestin, can suppress the hypothalamic-pituitary-gonadal axis but can also directly inhibit gonadal steroidogenesis; the success of MPA as a treatment for gonadotropin-independent sexual precocity derives from its direct action on steroidogenic tissues. Dexamethasone, a widely used glucocorticoid, can suppress the hypothalamic pituitary-adrenal axis, but its potential effect directly on the adrenal is unclear. Previous reports suggested that these two drugs may act on the initial steps in the rodent steroidogenic pathway; therefore, we investigated their abilities to inhibit the first three human enzymes in steroidogenesis: the cholesterol side-chain cleavage enzyme (P450scc), the 17alpha-hydroxylase/17,20 lyase (P450c17), and type II 3beta-hydroxysteroid dehydrogenase/isomerase (3betaHSDII). We found no effect of either drug on P450scc in intact human choriocarcinoma JEG-3 cells. Using microsomes from yeast expressing human P450c17 or microsomes from human adrenals, we found that dexamethasone inhibited P450c17 with a Ki of 87 micromol/L, which is about 1000 times higher than typical therapeutic concentrations, but that MPA has no detectable action on P450c17. Using microsomes from yeast expressing human 3betaHSDII, we found that this enzyme has indistinguishable apparent Km values of 5.2-5.5 micromol/L and similar maximum velocities of 0.34-0.56 pmol steroid/min x microg microsomal protein for the three principal endogenous substrates, pregnenolone, 17-hydroxypregnenolone, and dehydroepiandrosterone. In this system, MPA inhibited 3betaHSDII with a Ki of 3.0 micromol/L, which is near concentrations achieved by high therapeutic doses of 5-20 mg MPA/kg x day. These data establish the mechanism of action of MPA as an inhibitor of human steroidogenesis, and are in contrast with the results of earlier studies indicating that MPA inhibited both P450c17 and 3betaHSD in rat Leydig cells. These studies establish the "humanized yeast" system as a model for studying the actions of drugs on human steroidogenic enzymes and suggest that 3betaHSDII may be an appropriate target for pharmacological interventions in human disorders characterized by androgen excess or sex steroid dependency. PMID- 10372720 TI - Analysis of carbohydrate residues on recombinant human thyrotropin receptor. AB - An investigation of the sugar groups on recombinant human TSH receptors (TSHR) expressed in CHO-K1 cells and solubilized with detergents is described. Western blotting studies with TSHR monoclonal antibodies showed that the receptor was present principally as two bands with approximate molecular masses of 120 and 100 kDa. Further blotting studies using lectins and/or involving treatment with different glycosidases indicated that the 100-kDa band contained about 16 kDa of high mannose-type sugars, and the 120-kDa band contained about 33 kDa of complex type sugars. It was possible to separate the 120- and 100-kDa components of the TSHRs by lectin affinity chromatography. In particular, Galanthus nivalis lectin, which binds high mannose-type sugars, bound the 100-kDa band, but not the 120-kDa band, whereas Datura stramonium lectin, which binds complex-type sugars, bound the 120-kDa band, but not the 100-kDa band. 125I-Labeled TSH binding studies with the various lectin column fractions showed that TSH-binding activity was principally associated with the complex-type sugar containing the 120-kDa form of the receptor rather than the high mannose-containing 100-kDa form. During peptide chain glycosylation, high mannose-type sugar residues are attached first and then modified by the formation of complex type structures to form the mature glycoprotein. Our data suggest that in the case of the TSH receptor, this type of posttranslational processing has an important role in forming the TSH-binding site. PMID- 10372721 TI - The differential effect of food intake and beta-adrenergic stimulation on adipose derived hormones and cytokines in man. AB - We determined whether the physiologic changes that accompany food intake or sympathetic activation by beta-adrenergic stimulation result in alterations in the secretion of leptin, tumor necrosis factor-alpha (TNF alpha), or interleukin 6 (IL-6) by serially sampling sc abdominal adipose interstitial fluid by open flow microperfusion before and after a standardized meal and in response to isoproterenol (1 micromol/L) delivered locally. Post cibum IL-6 rose up to 5 fold, whereas leptin and TNF alpha secretion did not change; TNF alpha, but not IL-6, correlated positively with indices of lipolysis. Isoproterenol-induced lipolysis was accompanied by a transient 40% reduction in leptin and a parallel 85% elevation of TNF alpha concentration, whereas IL-6 levels did not change; again, TNF alpha correlated positively with lipolysis. These data show that secretion of some, but not all, metabolically relevant polypeptides by adipose tissue is modulated within a short time frame by food or stress stimuli, suggesting a role of these peptides in local autocrine/paracrine or distant endocrine effects on fat metabolism. TNF alpha's close correlation with lipolysis suggests that this cytokine participates in a local positive autocrine feedback loop, potentiating lipolysis and inhibiting insulin's antilipolytic actions. The regulations of adipose leptin, TNF alpha, and IL-6 secretion seem distinct from each other and different in the fed vs. fasting state. PMID- 10372722 TI - Inhibition of growth hormone (GH) secretion by a mutant GH-I gene product in neuroendocrine cells containing secretory granules: an implication for isolated GH deficiency inherited in an autosomal dominant manner. AB - Isolated GH deficiency (IGHD) type II is a disease inherited in an autosomal dominant manner. Although point mutations at the donor splice site of intron 3 of the GH-I gene have been identified in patients, the mechanism of how such mutations result in severe GH deficiency is unclear. Recently, we identified two mutations in Japanese patients with IGHD type II, G to A substitutions at the first (mutA) and fifth (mutE) nucleotides of intron 3. Messenger ribonucleic acids skipping exon 3 were transcribed from both mutant GH-I genes. We studied in this report the synthesis and secretion of GH encoded by the mutant GH-I genes and tested whether inhibition of wild-type GH secretion by mutant products could be demonstrated in cultured cell lines. A metabolic labeling study in COS-1 cells revealed that a mutant GH with a reduced molecular mass was synthesized from the mutant messenger ribonucleic acid and retained in the cells for at least 6 h. On the other hand, the wild-type GH was rapidly secreted into the medium. Coexpression of mutant and wild-type GH did not result in any inhibition of wild type GH secretion in COS-1 or HepG2 cells. However, coexpression of mutant GH resulted in significant inhibition of wild-type GH secretion in somatotroph derived MtT/S cells as well as in adrenocorticotroph-derived AtT-20 cells, without affecting cell viability. We conclude that the dominant negative effect of mutant GH on the secretion of wild-type GH is at least in part responsible for the pathogenesis of IGHD type II. Our results also suggest that neuroendocrine cell type-specific mechanisms, including intracellular storage of the secretory proteins, are involved in the inhibition. PMID- 10372723 TI - Tripartite neuroendocrine activation of the human growth hormone (GH) axis in women by continuous 24-hour GH-releasing peptide infusion: pulsatile, entropic, and nyctohemeral mechanisms. AB - Despite the discovery of potent GH-releasing peptides (GHRPs) more than 15 yr ago and the recent cloning of human, rat, and pig GHRP receptors in the hypothalamus and pituitary gland, the neuroregulatory mechanisms of action of GHRP agonists on the human hypothalamo-somatotroph unit are not well delineated. To gain such clinical insights, we evaluated the ultradian (pulsatile), entropic (pattern orderliness), and nyctohemeral GH secretory responses during continuous 24-h i.v. infusion of saline vs. the most potent clinically available hexapeptide, GHRP-2 (1 microg/kg x h) in estrogen-unreplaced (mean serum estradiol, 12 +/- 2.4 pg/mL) postmenopausal women (n = 7) in a paired, randomized design. Blood was sampled every 10 min for 24 h during infusions and was assayed by ultrasensitive GH chemiluminescence assay. Pulsatile GH secretion was quantitated by deconvolution analysis, orderliness of GH release patterns by the approximate entropy statistic, and 24-h GH rhythmicity by cosinor analysis. Statistical analysis revealed that GHRP-2 elicited a 7.7-fold increase in (24-h) mean serum (+/-SEM) GH concentrations, viz. from 0.32 +/- 0.042 (saline) to 2.4 +/- 0.34 microg/L (GHRP-2; P = 0.0006). This occurred via markedly stimulated pulsatile GH release, namely a 7.1-fold augmentation of GH secretory burst mass: 0.87 +/- 0.18 (control) vs. 6.3 +/- 1.3 microg/L (GHRP-2; P = 0.0038). Enhanced GH pulse mass reflected a commensurate 10-fold (P = 0.023) rise in GH secretory burst amplitude [maximal GH secretory rate (micrograms per L/min) attained within a secretory pulse] with no prolongation in event duration. GH burst frequency, interpulse interval, and calculated GH half-life were all invariant of GHRP-2 treatment. Concurrently, as detected in the ultrasensitive GH assay, GHRP-2 augmented deconvolution-estimated interpulse (basal) GH secretion by 4.5-fold (P = 0.025). The approximate entropy of 24-h serum GH concentration profiles rose significantly during GHRP-2 infusion; i.e. from 0.592 +/- 0.073 (saline) to 0.824 +/- 0.074 (GHRP-2; P = 0.0011), signifying more irregular or disorderly GH release patterns during secretagogue stimulation. Cosinor analysis of 24-h GH rhythms disclosed a significantly earlier (daytime) acrophase at 2138 h (+/- 140 min) during GHRP-2 stimulation vs. 0457 h (+/-42 min) during saline infusion (P = 0.013). Concomitantly, the cosinor amplitude rose 6-fold (P = 0.018), and the mesor (cosine mean) rose 5-fold (P = 0.003). Fasting (0800 h) plasma insulin-like growth factor (IGF-I) concentrations rose by -11 +/- 12 microg/L during saline infusion and by 102 +/- 18 microg/L during GHRP-2 infusion (P = 0.0036). GHRP-2 infusion did not modify (24-h pooled) serum LH, FSH, or TSH concentrations and minimally increased serum (pooled) daily PRL (6.8 +/- 0.83 vs. 12 +/- 1.2 microg/L; P < 0.05) and cortisol (5.3 +/- 0.59 to 7.0 +/- 0.74; P < 0.05) concentrations. In summary, 24-h constant iv GHRP-2 infusion in the gonadoprival female neurophysiologically activates the GH-IGF-I axis by potentiating GH secretory burst mass and amplitude by 7- to 10-fold and augmenting the basal (nonpulsatile) GH secretion by 4.5-fold. GHRP-2 action is highly selective, as it does not alter GH secretory burst frequency, interpulse interval, event duration, or GH half-life. GHRP-2 effectively elevates IGF-I concentrations, unleashes greater disorderliness of GH release patterns, and heightens the 24-h rhythmicity of GH secretion. These tripartite features of GHRP-2's action in estrogen withdrawn (postmenopausal) women also characterize normal human puberty and/or sex steroid regulation of the GH-IGF-I axis. However, how or whether GHRP-2 interacts further with sex hormone modulation of GH neurosecretory control in older women and men is not yet known. PMID- 10372724 TI - Effects of gonadal suppression on the regulation of parathyroid hormone and 1,25 dihydroxyvitamin D secretion in women. AB - Although a causal association between estrogen deficiency and bone loss has been established for many years, the mechanism by which estrogen deficiency leads to bone loss is unclear. Estrogen deficiency could induce bone loss either by a direct effect on bone cells to modify the production of bone-resorbing cytokines or by altering the production or response to calcium regulatory hormones such as PTH and 1,25-dihydroxyvitamin D. To assess the effects of ovarian hormones on calcium regulatory hormones, we evaluated the ability of calcium to suppress PTH secretion and the ability of PTH to increase serum 1,25-dihydroxyvitamin D and whole blood ionic calcium levels in women before and after GnRH analog-induced ovarian suppression. Sixteen women with endometriosis underwent i.v. infusion of calcium (1.1 mg calcium gluconate/cc in 5% dextrose) at a rate of 4 cc/kg x h (n = 7) or human PTH-(1-34) (Parathar) at a dose of 0.55 U/kg x h (n = 9) before and after 6 months of suppression of ovarian function with the GnRH analog nafarelin acetate (200 microg, intranasally, twice daily). Initial infusions were performed between days 6-10 of the menstrual cycle. Serum PTH and whole blood ionic calcium levels were measured at -20, -10, and 0 min and then every 10 min for 2 h during i.v. calcium infusions. Whole blood ionic calcium and 1,25-dihydroxyvitamin D levels were measured every 6 h for 24 h during i.v. human PTH-(1-34) infusions. Serum estradiol levels were markedly suppressed by nafarelin therapy in both groups of women. The relationship between whole blood ionic calcium and serum PTH levels was similar before and during nafarelin-induced ovarian suppression. The net change and rate of rise in serum 1,25-dihydroxyvitamin D levels in response to PTH infusion were similar before and during nafarelin therapy. Peak whole blood ionic calcium and incremental increases in ionic calcium in response to PTH were similar before and during nafarelin therapy. Our data suggest that ovarian suppression does not alter the regulation of PTH secretion in response to calcium, the ability of PTH to stimulate 1,25-dihydroxyvitamin D formation, or the skeletal sensitivity to PTH. These findings suggest that alterations in calcium regulatory hormones by estrogen deficiency are unlikely to play a major role in the pathogenesis of estrogen deficiency bone loss. PMID- 10372725 TI - Genetic heterogeneity in familial nonmedullary thyroid carcinoma: exclusion of linkage to RET, MNG1, and TCO in 56 families. NMTC Consortium. AB - Epidemiological studies show a very high relative risk for first degree relatives of probands with thyroid cancer. The familial form of nonmedullary thyroid carcinoma (NMTC) gives a more severe phenotype and appears earlier than its sporadic counterpart. Moreover, benign thyroid pathologies are often observed in NMTC kindreds. Little is known about the genetic risk factors of the disease. To study them, an international consortium has been organized at the International Agency for Research on Cancer over the past 2 yr to collect biological samples from NMTC families. The only genes known to be directly involved in susceptibility to NMTC are MNG1 on chromosome 14q32 and TCO on chromosome 19q13.2, previously localized by us and others. In addition to those two genes, the genes for Cowden's syndrome and familial adenomatous polyposis are associated with thyroid cancer, but not as an indicative phenotype. Another important gene in thyroid carcinogenesis is RET, which is mutated in the majority of cases of hereditary medullary thyroid cancer and rearranged in an important fraction of sporadic cases of NMTC. Here we report the result of a linkage analysis performed on the 56 more informative kindreds we have collected through the international consortium. Linkage analysis using both parametric and nonparametric methods excluded MNG1, TCO, and RET as major genes of susceptibility to NMTC and demonstrated that this trait is characterized by genetic heterogeneity. PMID- 10372726 TI - Inhibin A and inhibin B responses to gonadotropin withdrawal depends on stage of follicle development. AB - Previous studies demonstrate that inhibin A and B are differentially secreted across the menstrual cycle. To test the hypothesis that the responses of inhibin A and inhibin B to partial gonadotropin withdrawal are influenced by the stage of follicular development, a maximally suppressive dose of the Nal-Glu GnRH antagonist (150 microg/kg) was administered to normal cycling women during the midfollicular (MFP; n = 8), late follicular (LFP; n = 6), and midluteal phase (MLP; n = 5) to assess ovarian responsiveness over a broad range of developmental stages. Administration of the GnRH antagonist resulted in a significant decrease in LH (75 +/- 5%, 63 +/- 3%, and 84 +/- 7%; P < 0.05) and FSH (23 +/- 9%, 32 +/- 5%, and 39 +/- 6%; P < 0.04) during the MFP, LFP, and MLP, respectively. During the MFP, partial withdrawal of gonadotropins resulted in disappearance of the dominant follicle on ultrasound accompanied by a decrease in estradiol (E2; 64.9 +/- 11.4 to 23.9 +/- 3.3 pg/mL; P < 0.02) and inhibin B levels (121.6 +/- 14.8 to 28.4 +/- 4.8 pg/mL; P < 0.002) from baseline to near the limit of detection. Inhibin A was near the limit of detection in this cycle stage (0.8 +/- 0.1 IU/mL). When gonadotropins were withdrawn during the LFP, the size of the dominant follicle remained stationary in four of five subjects, and inhibin B (84.1 +/- 14.1 to 22.2 +/- 3.4 pg/mL; 71 +/- 5%; P < 0.02), inhibin A (4.4 +/- 1.1 to 1.3 +/- 0.5 IU/mL; 71 +/- 7%; P < 0.02), and E2 (236.8 +/- 48.2 to 95.6 +/ 39.0 pg/mL; 61 +/- 12%; P < 0.02) decreased similarly. Time to ovulation was shorter in association with higher inhibin A (r = -0.67; P < 0.02) and E2 (r = 0.79; P < 0.003), but there was no relation to inhibin B. During the MLP, decreased gonadotropin levels resulted in the disappearance of corpus luteum function with a significant decrease in inhibin A (9.0 +/- 0.4 to 0.7 +/- 0.1 IU/mL; 92 +/- 1%; P < 0.0001) in combination with decreased E2 (150.4 +/- 22.9 to 23.8 +/- 4.2 pg/mL; 83 +/- 3%; P < 0.005) and progesterone (12.6 +/- 2.6 to 0.9 +/- 0.2 ng/mL; 92 +/- 2%; P < 0.01). Administration of a GnRH antagonist at precise stages of the menstrual cycle provides further evidence that differential regulation of inhibin A and inhibin B is critically dependent on the stage of follicular development. Inhibin B secretion is exquisitely sensitive to gonadotropin withdrawal during the MFP when inhibin A has not yet risen. Inhibin A and inhibin B decrease equally after GnRH antagonist administration during the LFP. However, before antagonist administration, the positive correlation between estradiol and inhibin A and time to ovulation and the lack of such a correlation with inhibin B suggest that the source of inhibin B secretion is different from that of inhibin A and E2. The decrease in inhibin A secretion after antagonist administration during the luteal phase confirms gonadotropin-dependent secretion of dimeric inhibin A by the corpus luteum. PMID- 10372727 TI - Biotransformation of oral dehydroepiandrosterone in elderly men: significant increase in circulating estrogens. AB - The most abundant human steroids, dehydroepiandrosterone (DHEA) and its sulfate ester DHEAS, may have a multitude of beneficial effects, but decline with age. DHEA possibly prevents immunosenescence, and as a neuroactive steroid it may influence processes of cognition and memory. Epidemiological studies revealed an inverse correlation between DHEAS levels and the incidence of cardiovascular disease in men, but not in women. To define a suitable dose for DHEA substitution in elderly men we studied pharmacokinetics and biotransformation of orally administered DHEA in 14 healthy male volunteers (mean age, 58.8 +/- 5.1 yr; mean body mass index, 25.5 +/- 1.5 kg/m2) with serum DHEAS concentrations below 4.1 micromol/L (1500 ng/mL). Diurnal blood sampling was performed on 3 occasions in a single dose, randomized, cross-over design (oral administration of placebo, 50 mg DHEA, or 100 mg DHEA). The intake of 50 mg DHEA led to an increase in serum DHEAS to mean levels of young adult men, whereas 100 mg DHEA induced supraphysiological concentrations [placebo vs. 50 mg DHEA vs. 100 mg DHEA; area under the curve (AUC) 0-12 h (mean +/- SD) for DHEA, 108 +/- 22 vs. 252 +/- 45 vs. 349 +/- 72 nmol/L x h; AUC 0-12 h for DHEAS, 33 +/- 9 vs. 114 +/- 19 vs. 164 +/- 36 micromol/L x h]. Serum testosterone and dihydrotestosterone remained unchanged after DHEA administration. In contrast, 17beta-estradiol and estrone significantly increased in a dose-dependent manner to concentrations still within the upper normal range for men [placebo vs. 50 mg DHEA vs. 100 mg DHEA; AUC 0-12 h for 17beta-estradiol, 510 +/- 198 vs. 635 +/- 156 vs. 700 +/- 209 pmol/L x h (P < 0.0001); AUC 0-12 h for estrone, 1443 +/- 269 vs. 2537 +/- 434 vs. 3254 +/- 671 pmol/L x h (P < 0.0001)]. In conclusion, 50 mg DHEA seems to be a suitable substitution dose in elderly men, as it leads to serum DHEAS concentrations usually measured in young healthy adults. The DHEA-induced increase in circulating estrogens may contribute to beneficial effects of DHEA in men. PMID- 10372728 TI - Post-translational processing of the natural human thyrotropin receptor: demonstration of more than two cleavage sites. AB - Epitope-mapped monoclonal and polyclonal antibodies to the TSH receptor (TSHR) were used as immunoblot probes to detect and characterize the molecular species of the receptor present in normal human thyroid tissue. In reduced membrane fractions, both full-length (uncleaved) holoreceptor and cleavage-derived subunits of the holoreceptor were detected. Uncleaved holoreceptor species included a nonglycosylated form of apparent molecular mass 85 kDa and two glycosylated forms of approximately 110 and 120 kDa. The membranes also contained several forms of cleavage-derived TSHR-alpha and TSHR-beta subunits. TSHR-alpha subunits were detected by antibodies to epitopes localized within the amino terminal end of the TSHR ectodomain and migrated diffusely between 45-55 kDa, reflecting a differentially glycosylated status. TSHR-beta subunits were detected by antibodies to epitopes within the carboxyl end of the TSHR ectodomain. Several species of TSHR-beta subunit were present, the most abundant having apparent molecular masses of 50, 40, and 30 kDa. These data demonstrated that post translational processing of the TSHR in human thyroid tissue involved multiple cleavage sites. PMID- 10372729 TI - Divergent effects of weight reduction and oral anticonception treatment on adrenergic lipolysis regulation in obese women with the polycystic ovary syndrome. AB - The influence of weight reduction and female sex hormones on the regulation of lipolysis was investigated in isolated abdominal sc adipocytes from 20 obese hyperandrogenic women with polycystic ovary syndrome (PCOS). Nine PCOS women were reinvestigated after 8-12 weeks of weight reduction therapy (WR) with a very low calorie diet, inducing a mean loss of 8 +/- 3 kg, and 8 PCOS women were reinvestigated after 12 weeks of treatment with combined oral contraceptives (OC), containing ethinyl estradiol and norethisterone; the remaining 3 subjects were drop-outs. Both WR and OC normalized hyperandrogenicity. WR caused a 50% reduction of basal lipolysis rate and a 5- to 7-fold increased noradrenaline and terbutaline sensitivity (P < 0.02); the latter could be ascribed to a 2-fold increased beta2-adrenoceptor density (P < 0.02) as determined with radioligand binding. There was no change with regard to dobutamine (beta1-adrenoceptor sensitivity) or clonidine, (alpha2-adrenoceptor sensitivity) or to beta1 adrenoceptor density. OC treatment did not influence the basal lipolysis rate or beta2- or alpha2-adrenoceptor sensitivity, but lowered the beta1-adrenoceptor sensitivity 7-fold (P < 0.03) without a reduction in beta1-adrenoceptor density. The OC treatment effect was not observed when forskolin and dibutyryl cAMP, acting on adenylate cyclase or protein kinase A, respectively, were used, suggesting a partial uncoupling of beta1-adrenoceptors. WR therapy, but not OC therapy, caused, in addition to changes in lipolysis function, improved in vivo insulin sensitivity and lower plasma noradrenaline levels. These findings suggest that factors other than hyperandrogenicity modulate lipolysis regulation in obese subjects with PCOS. Disturbances in sympathetic pathways could be of pathogenic importance. PMID- 10372730 TI - Placental Fas ligand expression is a mechanism for maternal immune tolerance to the fetus. AB - Fas ligand (FasL) is a peptide that plays an important immunoregulatory role in limiting the host immune response. Several studies have shown that the expression of FasL in the anterior chamber of the eye and the testis allows these tissues to be immunoprivileged sites. Immunotolerance is achieved by binding of FasL to its receptor (Fas) on activated immune cells, which results in cell apoptosis. To determine whether FasL has a role in maternal immune tolerance to the fetus, we looked for the expression of FasL in the human placenta. Immunoperoxidase staining localized FasL to both syncytiotrophoblast and cytotrophoblast in placental villi and chorionic extravillous trophoblast. Western analysis demonstrated FasL in placental villi and a human first-trimester trophoblast cell line (ED27). In contrast, Fas was colocalized to CD45 (leukocyte common antigen) positive cells found in maternal decidua. When isolated peripheral blood lymphocytes were induced to express Fas with phytohemagglutinin (PHA) and interleukin-2 (IL-2) and then cocultured with trophoblast, 30% of the lymphocytes underwent apoptosis, as determined by the in situ death (TUNEL) assay. Neutralizing antibodies to FasL inhibited apoptosis by 40% in these studies. In contrast, activated lymphocytes cocultured with non-FasL-expressing fibroblasts or unactivated non-Fas-expressing lymphocytes cocultured with ED27 trophoblast showed little evidence of apoptosis. These findings suggest that FasL expressed by fetal trophoblast cells can induce apoptosis in activated lymphocytes there by providing a mechanism for maternal immune tolerance to the fetus. PMID- 10372731 TI - Vasopressin receptors in human adrenal medulla and pheochromocytoma. AB - The nature of vasopressin (VP) receptors present in normal and tumoral human adrenal was investigated using various experimental approaches. Specific VP binding sites were detected by autoradiography using [3H]arginine VP as a radioligand in adrenal cortex and medulla. The V1a receptor subtype was expressed in the two parts of the gland, as shown by pharmacological studies and RT-PCR experiments. By contrast, the V1b receptor subtype was only expressed in medullary chromaffin cells. This was confirmed by the characterization of V1b transcripts detected in adrenal medulla tissues. In pheochromocytoma, we also detected functional V1b receptors. These receptors triggered intracellular calcium mobilization from intracellular pools and were involved in catecholamine secretion. Binding experiments performed on pheochromocytoma plasma membrane preparations also revealed V1a vasopressin-binding sites, whose roles and cellular localization have not yet been determined. RT-PCR experiments confirmed these data; 100% and 80% of the five tumors tested exhibited V1a and V1b transcripts, respectively. Perifusion experiments also demonstrated that some pheochromocytomas may secrete large amounts of VP. Our findings imply that VP locally secreted by human adrenal medulla may regulate adrenal function by acting on V1a or V1b receptors. More interestingly, we demonstrate that one pheochromocytoma oversecretes VP. In this particular case, this may contribute to the increase in blood pressure observed. PMID- 10372732 TI - Basal and interleukin-1beta-stimulated prostaglandin production from cultured human myometrial cells: differential regulation by corticotropin-releasing hormone. AB - During pregnancy, placental CRH acts on human myometrium via specific receptors and might play a role in the regulation of myometrial contractility and hence human parturition. The myometrium is the site of production and target of several PGs, which can be activated by cytokines, especially during infection-induced preterm labor. We established primary human myometrial cell cultures that express functional CRH receptors (CRH-R1alpha, -R1beta, -Rlc, and -R2beta) to investigate the possible regulation of PG production by CRH. We studied the effect of CRH on the two major myometrial PGs, PGE2 and 6-keto PGF1alpha. Human CRH was able to partially inhibit basal, interleukin-1beta-stimulated, and oxytocin-stimulated PGE2 production (56 +/- 11%, 45 +/- 8%, and 58 +/- 6% inhibition, respectively). This effect was blocked by a specific CRH receptor antagonist in a concentration dependent manner. Furthermore, CRH had no effect on 6-keto PGF1alpha production, indicating that the CRH inhibitory action does not involve suppression of cyclooxygenase, the enzyme responsible for the production of both PGE2 and 6-keto PGF1alpha. These data further support the view that during pregnancy, CRH may promote myometrial quiescence and might play an important role in the regulation of human labor. PMID- 10372733 TI - Expression of the members of the Ptx family of transcription factors in human pituitary adenomas. AB - A number of putative transcription factors described in the pituitary have been implicated as key elements in the processes that direct pituitary development. Three recently described proteins, Ptx1, Ptx2, and Ptx3, define a new family of transcription factors, the Ptx subfamily, within the paired-like class of homeodomain factors. In mice, Ptx1 and Ptx2 gene expression has been detected in the area of the pituitary primordium and is maintained throughout development in Rathke pouch and adult pituitary. In the present study, the expression of the Ptx1, Ptx2, and Ptx3 genes was characterized in the normal human pituitary and in the different types of human pituitary adenomas. Although no Ptx3 gene expression could be detected in these tissues, Ptx1 presented with a quite ubiquitous pattern of distribution, being expressed at quite constant levels in normal tissues and in all 60 pituitary tumors analyzed. The pattern of expression of the Ptx2 gene among the different subsets of pituitary adenomas was even more varied. No Ptx2 expression could be detected in corticotroph tumors. In contrast, high levels of Ptx2 messenger ribonucleic acid were measured in the gonadotroph tumors, although no specific correlation to other markers of the gonadotroph lineage differentiation, such as alphaGsu, LHbeta, or FSHbeta, could be evidenced. Finally, Ptx2 was also expressed in pure lactotroph adenomas and not in somatotroph adenomas. Ptx2 is, therefore, the first paired homeodomain pituitary transcription factor differentially expressed in these two lineages, which derive from a common precursor. These results support a role for Ptx2 in the terminal differentiation of somatotroph and lactotroph cell phenotypes. PMID- 10372734 TI - Immunohistochemical analysis of Bcl-2, Bax, and Bak expression in thyroid glands from patients with subacute thyroiditis. AB - Bcl-2 family proteins are important regulators of apoptosis. To clarify a role of apoptosis and the expression of Bcl-2 family proteins in the pathogenesis of subacute thyroiditis (SAT), we evaluated the expression of Bcl-2, Bax, and Bak by immunohistochemistry and apoptosis by in situ end labeling of fragmented DNA in thyroid tissues from 11 patients with SAT. Apoptotic nuclei were found in granulomas, especially in macrophages/histiocytes and lymphocytes, and in the regenerating follicular cells, but were rarely found in the area of fibrosis. The mean (+/-SD) percentage of apoptotic follicular cells was significantly greater in SAT than that in controls (1.4 +/- 0.8% vs. 0.4 +/- 0.6%). Bcl-2, Bak, and Bax were strongly expressed in the granulomas and regenerating thyroid follicular cells from patients with SAT. Bcl-2 and Bak, but not Bax, were expressed in follicular cells from normal controls. The percentage of apoptotic cells and the expression of Bax in follicular cells did not correlate with age or serum levels of thyroid hormones, C-reactive protein, or thyroglobulin. These data suggest that apoptosis may be involved in the development of SAT and that Bax expression in regenerating thyrocytes may be important for the recovery of SAT. PMID- 10372735 TI - Endometrial and myometrial expression of nitric oxide synthase isoforms in pre- and postmenopausal women. AB - Expression of nitric oxide synthase (NOS) protein was examined by Western immunoblot analysis and immunohistochemistry in the endometrium and myometrium of 19 premenopausal and 18 postmenopausal women undergoing hysterectomy for benign gynecological reasons. The predominant isoform of NOS in the human uterus was endothelial NOS (eNOS). Using immunohistochemistry, eNOS was localized predominantly to the glandular epithelium and endometrial microvascular endothelium. eNOS was scant and inconsistently detected in endometrial stromal cells. In the myometrium, eNOS was predominantly found in smooth muscle cells (myocytes) and was also detected in the microvascular endothelium. Neuronal NOS was not detectable by immunohistochemical techniques, and inducible NOS (iNOS) was only detectable in occasional specimens, although more often in secretory specimens. iNOS, when present, was predominantly found in glandular epithelium and occasional stromal cells. Myometrial iNOS was scant and not consistently detected. By Western immunoblot analysis, neuronal NOS or iNOS was not detected. We observed a unique menstrual cycle-dependent expression of eNOS that was different in the endometrium compared to the myometrium and was independent of uterine pathology. In the endometrium, there was 62% higher expression of eNOS during the secretory phase (P = 0.00085) compared to the proliferative phase, whereas in the myometrium, there was 74% greater expression of eNOS in the proliferative phase (P = 0.03) compared to the secretory phase. Within the secretory phase, maximal endometrial eNOS expression was found in the midportion, whereas in the myometrium, highest eNOS expression occurred during the late secretory phase. In postmenopausal women not treated with hormones, a significant reduction in endometrial and myometrial expression of eNOS occurred, which was reversed by continuous hormone replacement therapy. In summary, both endogenous ovarian steroids and exogenous sex hormones influence uterine eNOS expression. Our results suggest that estrogen may regulate myometrial eNOS, whereas progesterone or a combination of estrogen and progesterone may be more important in regulating endometrial eNOS, and NO may be a critical mediator of sex steroid actions in the human uterus. PMID- 10372736 TI - Analysis of mutations in genes of the follicle-stimulating hormone receptor signaling pathway in ovarian granulosa cell tumors. AB - It has been suggested that ovarian granulosa cell tumors may result from unopposed hyperstimulation, either by excessive gonadotropin stimulation, by activating mutations of the FSH receptor gene, or of the G protein subunits, Gs alpha or Gi alpha2. We have examined the entire open reading frame of the FSH receptor gene in ovarian granulosa cell tumors. In addition, these tumors were evaluated for the known oncogenic G protein mutations Gsp and Gip2. Normal results were obtained in all 23 ovarian granulosa cell tumors. We conclude that mutations of the FSH receptor G protein signaling pathway do not play any major role in the genesis of these tumors. PMID- 10372738 TI - Desmopressin normalizes the blunted adrenocorticotropin response to corticotropin releasing hormone in melancholic depression: evidence of enhanced vasopressinergic responsivity. AB - Major depression is associated with significant disturbance in hypothalamic pituitary-adrenal axis functioning, including blunted release of ACTH in response to CRH infusion. Eight melancholic depressives and eight matched healthy comparison subjects underwent, in random order, the following challenges: placebo, CRH, CRH + DDAVP. Blood for ACTH and cortisol estimation was drawn at 15, 0, 15, 30, 45, 60, 90, and 120 min. A blunted release of ACTH, in response to CRH challenge, was observed in depression (P < 0.01), whereas maximal cortisol responses in both groups were similar, despite elevated baseline levels in depression (P < 0.05). The combined CRH/DDAVP infusion produced similar ACTH and cortisol release in both groups. These results suggest that melancholic depression is associated with enhanced pituitary vasopressinergic responsivity. PMID- 10372737 TI - Urinary excretion of aquaporin-2 water channel differentiates psychogenic polydipsia from central diabetes insipidus. AB - The present study was undertaken to determine whether urinary excretion of aquaporin-2 (AQP-2) water channel under ad libitum water intake is of value to differentiate polyuria caused by psychogenic polydipsia from central diabetes insipidus. A 30-min urine collection was made at 0900 h in 3 groups of: 11 patients with central diabetes insipidus (22-68 yr old), 10 patients with psychogenic polydipsia (28-60 yr old), and 15 normal subjects (21-38 yr old). In the patients with central diabetes insipidus, the plasma arginine vasopressin level was low despite hyperosmolality, resulting in hypotonic urine. Urinary excretion of AQP-2 was 37 +/- 15 fmol/mg creatinine, a value one-fifth less than that in the normal subjects. In the patients with psychogenic polydipsia, plasma arginine vasopressin and urinary osmolality were as low as those in the patients with central diabetes insipidus. However, urinary excretion of AQP-2 of 187 +/- 45 fmol/mg creatinine was not decreased, and its excretion was equal to that in the normal subjects. These results indicate that urinary excretion of AQP-2, under ad libitum water drinking, participates in the differentiation of psychogenic polydipsia from central diabetes insipidus. PMID- 10372739 TI - Discovery of a Met300Val variant in Shc and studies of its relationship to birth weight and length, impaired insulin secretion, insulin resistance, and type 2 diabetes mellitus. AB - The Shc adaptor proteins corresponding to the 46-, 52-, and 66-kDa isoforms are key transducers of growth promotion and gene expression, which are being phosphorylated by all known receptor tyrosine kinases after stimulation by growth factors such as insulin and insulin-like growth factor I. Several studies have demonstrated a relationship between intrauterine growth retardation and impaired glucose tolerance or type 2 diabetes later in life. It is unclear whether this finding is partially explained by genetic factors. In this context, abnormalities in Shc proteins are considered to be a plausible candidate. Therefore, the aim of this study was to analyze whether genetic variability of the Shc isoforms causes a decrease in cell growth and cell differentiation that could be manifested by a decrease in birth weight and length, impaired acute insulin secretion after i.v. glucose, insulin resistance, and eventually a higher prevalence of type 2 diabetes. By single strand conformation polymorphism-heteroduplex analysis of 70 patients with diabetes mellitus and subsequent nucleotide sequencing of identified single strand conformation polymorphism variant, we discovered a Met300Val substitution of the 52-kDa isoform. The amino acid variant was predicted to be present in all 3 isoforms of Shc. In a genotype-phenotype study of 360 young healthy subjects, the allelic frequency of the codon 300 polymorphism was 4.2%. In this cohort, no significant differences could be shown between carriers and noncarriers in birth weight and length, the acute insulin response to i.v. glucose, or the insulin sensitivity index, as estimated from an i.v. glucose tolerance test. In an association study of 313 type 2 diabetic patients and 226 matched glucose-tolerant subjects, there was no significant difference in allelic frequency of the Shc variant (5.1% in diabetic patients vs. 3.1% in control subjects; P = 0.11). In conclusion, by itself the Met300Val polymorphism of Shc has no major impact on birth weight and length, insulin sensitivity index, acute glucose-induced insulin secretion, or prevalence of random type 2 diabetes mellitus. PMID- 10372740 TI - Hypoxia and cAMP stimulate vascular endothelial growth factor (VEGF) in human endometrial stromal cells: potential relevance to menstruation and endometrial regeneration. AB - The human female reproductive tract shows unique cycle-specific changes in vascularization. Vascular endothelial growth factor (VEGF) is a specific vascular endothelial mitogen which is produced by human endometrium and is known to be regulated by steroid hormones. Vasoconstriction during menstruation leads to endometrial hypoxia, a possible stimulus for angiogenesis. In the current study we tested the hypothesis that hypoxia and cAMP, a known stimulus for endometrial decidualization, can induce VEGF in human endometrial stromal cells. Decidualized as well as non decidualized stromal cells from 6 patients were exposed to normoxia (20% oxygen) and hypoxia (2% oxygen) for up to 72h. VEGF levels were assessed by Northern analysis using a 605 bp BamHI fragment of the human VEGF cDNA, and hybridization signals were normalized to levels of 18S RNA. VEGF protein was determined by ELISA. Hypoxia stimulated VEGF mRNA in decidualized stromal cells by 10.2 fold at 48h compared to normoxic controls. VEGF protein increased 10 fold by 48h and increased further to 13 fold at 72h. In the presence of 2% oxygen VEGF mRNA in nondecidualized endometrial stromal cells was increased 1.2-8 fold between 2 and 72h of treatment. VEGF protein also increased 1.2-9 fold in this time period. cAMP regulated both VEGF mRNA and protein in non decidualized stromal cells. VEGF mRNA increased 2-4 fold in 2-72h and protein production showed a 2-6 fold increase. VEGF was seen to be regulated by both cAMP and hypoxia in an additive manner. These results demonstrate that both non decidualized and decidualized endometrial stromal cells respond to hypoxia with increasing levels of VEGF. 8Br-cAMP, which is shown to increase VEGF levels in endometrial cells per se, has an additive effect on VEGF production under hypoxic conditions. This effect may have physiologic and pathophysiologic relevance during the process of menstruation and in post menstrual endometrial repair and angiogenesis. PMID- 10372741 TI - Identification of a potent phytoestrogen in hops (Humulus lupulus L.) and beer. AB - The female flowers of the hop plant are used as a preservative and as a flavoring agent in beer. However, a recurring suggestion has been that hops have a powerful estrogenic activity and that beer may also be estrogenic. In this study, sensitive and specific in vitro bioassays for estrogens were used for an activity guided fractionation of hops via selective solvent extraction and appropriate HPLC separation. We have identified a potent phytoestrogen in hops, 8 prenylnaringenin, which has an activity greater than other established plant estrogens. The estrogenic activity of this compound was reflected in its relative binding affinity to estrogen receptors from rat uteri. The presence of 8 prenylnaringenin in hops may provide an explanation for the accounts of menstrual disturbances in female hop workers. This phytoestrogen can also be detected in beer, but the levels are low and should not pose any cause for concern. PMID- 10372742 TI - Pro-EPIL forms are present in amniotic fluid and maternal serum during normal pregnancy. AB - Recent observations based on immunohistochemistry and RT-PCR demonstrate that early placenta insulin-like peptide (EPIL), encoded by the INSL4 gene, is present in the placenta during gestation. In the present study, we report on the development of a specific immunoassay, entirely based on two monoclonal anti peptide antibodies (mAbs), and its use for the detection of pro-EPIL forms in biological fluids during pregnancy. One mAb directed against the C-connecting peptide was used to capture pro-EPIL forms and their binding was revealed by a radiolabeled anti-A chain mAb as the indicator. A composite synthetic peptide, encompassing the C- and A-domains, was utilized as the standard. Under these experimental conditions, the assay displays a sensitivity limit of 2 ng/mL. Pro EPIL molecular forms were detected in both amniotic fluid and maternal serum of pregnant women. At 12 to 16 weeks of pregnancy, the pro-EPIL level was higher in amniotic fluid (246 +/- 50.8 ng/mL) than in maternal serum (5 +/- 2.0 ng/mL). As gestation advanced, so the concentration of pro-EPIL forms decreased in amniotic fluid while its level increased in maternal serum. Interestingly, in amniotic fluid, the pattern of pro-EPIL concentration during pregnancy is very similar to that observed for human chorionic gonadotropin (hCG) and its free subunits. The pattern of serum pro-EPIL concentration is similar to that of the free alpha subunit. Together with our previous immunohistochemical observations, these results indicate that pro-EPIL is preferentially secreted by cytotrophoblasts in amniotic fluid and that the biosynthesis of hCG subunits and EPIL may be regulated by common pathways. Overall, our observations strongly suggest that EPIL may play a critical physiological role during embryonic and foetal development. PMID- 10372743 TI - Changes in plasma concentrations of leptin and body fat composition during weight restoration in anorexia nervosa. PMID- 10372744 TI - Comment on clinical and prognostic relevance of Ret-PTC activation in patients with papillary thyroid carcinoma. PMID- 10372745 TI - Lack of specificity of urinary free cortisol determinations: why does it continue? PMID- 10372746 TI - Comment--do glucocorticoids mediate the hypoglycemia-induced rise in insulin-like growth factor binding protein-1? PMID- 10372747 TI - Comment on dangerous dogmas in medicine: the nonthyroidal illness syndrome. PMID- 10372748 TI - Comment on dangerous dogmas in medicine--the nonthyroidal illness syndrome. PMID- 10372749 TI - TSH receptor mutation database. PMID- 10372750 TI - Investigations of hydrocortisone and fludrocortisone in the treatment of chronic fatigue syndrome. PMID- 10372751 TI - A toxicokinetic analysis in a patient with acute glufosinate poisoning. AB - Incidents of poisoning in humans caused by the ingestion of the glufosinate ammonium containing herbicides are gradually increasing in Japan. This poisoning is characterized by various neurological symptoms such as disturbances of consciousness, convulsions and apnea which appear after an asymptomatic interval of several hours. We studied the toxicokinetics of glufosinate in a patient with this poisoning successfully treated without extracorporeal hemopurification. A 65 year-old male ingested BASTA, which contains 20% w/v of glufosinate ammonium, about 300 ml, more than the estimated human toxic dose. Four and a half hours after ingestion, he showed speech ataxia and systemic tremor. He was prophylactically intubated before the occurrence of serious respiratory failure. After 5 days of artificial ventilation he was extubated and discharged without any sequelae. We studied the serial change of serum glufosinate concentration every 3-6 h and assessed the urinary excretion of glufosinate every 24 h. The absorbed amount of glufosinate was estimated from the cumulative excreted in urine. Toxicokinetic analysis was performed using the two-compartment model. The changes in serum glufosinate concentration exhibited T1/2alpha of 1.84 and T1/2beta of 9.59 h. The apparent distribution volume at beta-phase and the total body clearance were 1.44 l/kg and 86.6 ml/min, respectively. Renal clearance was estimated to be 77.9 ml/min. The indication for extracorporeal hemopurification for this poisoning has been discussed. PMID- 10372752 TI - Cardiovascular and neurological toxicity of venlafaxine. AB - 1. We report a case of venlafaxine overdose and describe pharmacokinetic data on drug disposition. 2. Case report. Serial venlafaxine levels were measured and drug half-life calculated and compared to data at therapeutic concentrations. Metabolite concentrations were also measured and the potential for toxicity described with reference to individual variation in such metabolism 3. Venlafaxine can cause significant cardiac and neurotoxicity. Its potential for causing such toxicity may be dependent on whether an individual has the extensive metaboliser cytochrome CYP2D6 phenotype or the poor metaboliser phenotype PMID- 10372753 TI - Biological monitoring exposure of workers from plant producing carbon electrodes: quantification of benzo[a]pyrene DNA-adducts in leukocytes, by a 32P postlabelling method and an immunoassay. AB - The levels of benzo[a]pyrene were monitored for blood DNA-benzo[a]pyrene adducts in 17 workers from a plant producing carbon electrodes, with high exposure to benzo[a]pyrene (575-902-1149 ng m(-3)). Two different techniques, a 32P postlabelling method and a competitive immunoassay using polyclonal antibodies obtained from rabbits immunised with DNA modified by benzo[a]pyrene-trans-7,8 dihydrodiol-9,10-epoxide were used. For each worker, urinary 1-hydroxypyrene, a potential indicator of exposure to polycyclic aromatic hydrocarbons, was measured. The effect of tobacco by urinary cotinine measurement was also considered. The postlabelling and immunoassay detection limits for DNA benzo[a]pyrene adducts were respectively 0.15 and 10 fmol 50 microg(-1) of DNA. The results obtained by the two methods demonstrated a good detection of DNA benzo[a]pyrene adducts, but no direct relationship between the quantity of adducts and the concentration of benzo[a]pyrene in air-borne was noted in the studied plant. The levels of DNA-benzo[a]pyrene adducts obtained by immunoassay were significantly higher than those obtained by the 32P-postlabelling (P < 0.001). For several workers, variations due to professional or non professional factors must be taken into account in interpreting the results. In conclusion, the two methods used proved very efficient in determining DNA-benzo[a]pyrene adducts, and may be useful in monitoring human exposure to known and previously unidentified environmental genotoxic agents. PMID- 10372754 TI - To what extent are biomonitoring data available in chemical risk assessment? AB - 1. Chemical risk assessment integrates the identification of hazards and the human exposure levels which can be established from external and/or internal exposure data. 2. The availability of biomonitoring and metabolism animal data, the skin penetration ability, and the existence of atmospheric threshold limit values were examined for twelve substances of the European first list of priority existing substances. This investigation was focused on workplace exposures and on urinary biomarkers of exposure. Appropriate biomonitoring data appeared to be available for two substances: styrene and trichloroethylene. Some biomonitoring research has been conducted on acrylonitrile, buta-1,3-diene, cyclohexane, 1,4 dichlorobenzene, hydrogen fluoride, 2-(2-methoxyethoxy)ethanol, however additional studies could be usefully carried out. No biomonitoring data are available for alkanes, C10-13, chloro; benzene, C10-13-alkyl derivatives; bis(pentabromophenyl)ether; diphenylether, octabromo-derivative. 3. It was concluded that in some cases, biomonitoring data are either lacking or scarce. This is rather surprising since the selection of the substances of the priority list was based on high tonnage, widespread use, extent of human exposure, and toxicological concern. The development of biomonitoring information could be helpful in assessing individual or population chemical exposure whatever the source and route, and would result in both more realistic and more accurate risk assessments. PMID- 10372755 TI - Lead and calcium transport in human erythrocyte. AB - In this paper we report the lead (Pb) and calcium (Ca) uptake by erythrocyte ghosts. In both cases the transport was carried out by a passive transport system with two kinetic components (Michaelis-Menten and Hill). Pb and Ca were capable of inhibiting the transport of the other metal in a non-competitive way. Under hyperpolarization, the uptakes of Ca and Pb were enhanced and the Michaelis Menten component prevailed. Both Ca and Pb uptakes were inhibited by N-ethyl maleimide to the same extent. These results indicate that Pb and Ca share the same permeability pathway in human erythrocytes and that this transport system is electrogenic. PMID- 10372756 TI - Antibody forming cell response to nickel and nickel-coated fly ash in rats. AB - The potential of nickel as nickel chloride, native fly ash and Ni-coated fly ash to alter pulmonary and systemic immune response was evaluated upon intratracheal (I/T) exposure of rats. The animals were sensitised with sheep red blood cells (SRBC) through I/T and intraperitoneal (I/P) routes. Nickel exposure resulted in a decrease in the number of antibody forming cells (AFC) in lung associated lymph nodes (LALN) and spleen. In rats exposed to native fly ash there was a reduction in the number of AFC in LALN but not in spleen. The results did not demonstrate any significant difference in the immunosuppression of fly ash and Ni-coated fly ash exposed rats. The decrease in AFC formation in Ni-coated fly ash exposed animals was of a lesser magnitude than in rats exposed to Ni-alone. PMID- 10372757 TI - Liver preneoplastic changes in mice treated with the herbicide fomesafen. AB - 1. Effect of the diphenyl ether herbicide fomesafen on liver preneoplastic changes and porphyrin biosynthesis was examined in male C57BL/6J mice (0.23% in the diet for 14 months) and ICR mice (0.3% in the diet for 50 weeks). Fomesafen treatment resulted in preneoplastic changes (liver nodules and foci of altered hepatocytes) in both strains, uroporphyria developed only in ICR mice. 2. Iron pretreatment (600 mg/kg as a single dose) accelerated the development of fomesafen-induced preneoplastic changes in both mouse strains. The number of foci containing altered hepatocytes, as well as the number and size of liver nodules, were increased in iron-pretreated animals. 3. A single injection of iron induced marked uroporphyria in C57BL/6J mice after 14 months (liver porphyrin content 102 nmol/g). This uroporphyria was further potentiated by fomesafen administration (208 nmol/g). 4. In ICR mice, liver histology was apparently normal after a 3 month recovery from fomesafen treatment (0.32% for 9 months). Liver porphyrin content (260 nmol/g) started to decrease immediately after fomesafen withdrawal, but was still significantly elevated after 3 months (5 nmol/g), as compared to controls (1 nmol/g). 5. It is concluded that the toxicological evaluation of fomesafen should focus on liver porphyrin biosynthesis. PMID- 10372758 TI - Variations in the distribution of okadaic acid in organs and biological fluids of mice related to diarrhoeic syndrome. AB - Okadaic acid (OA) is the main toxin produced by dinoflagellates which can accumulate in the hepatopancreas of mussels and cause diarrhetic shellfish poisoning in consumers. This toxin is also a tumour promoter and a specific potent inhibitor of protein phosphatases 1 and 2A. No specific target organ is known for this toxin. This study concerns the distribution of [3H]OA in organs and biological fluids of Swiss mice having received a single dose per os of AO (50 microg/kg). The determination of the toxin extracted from mouse organs 24 h after administration of [3H]OA and derivatised with 9-anthryldiazomethane (ADAM) before HPLC and fluorescent detection showed the highest concentration in intestinal tissue and stomach. This distribution was even more pronounced in intestinal tissue, when animal were given per os 90 microg/kg which induced diarrhoea. The high concentrations of [3H]OA in intestinal tissues and contents 24 h after administration demonstrates a slow elimination of OA. When the dose of OA was increased from 50-90 microg/kg, the concentrations of the toxin in the intestinal content and faeces increased proportionally. A good correlation was found between an increase of OA in the intestinal tissue and the diarrhoea in animals given 90 microg/kg orally. Moreover OA was present in liver and bile and in all organs including skin and also fluids. Altogether these results confirmed an enterohepatic circulation of OA as previously shown. These data also revealed that in acute OA intoxication the concentration of the toxin in the intestinal tissues reaches cytotoxic concentrations in accordance with the diarrhoea which is the main symptom of OA poisoning. PMID- 10372759 TI - Leap-frog is a robust algorithm for training neural networks. AB - Optimization of perceptron neural network classifiers requires an optimization algorithm that is robust. In general, the best network is selected after a number of optimization trials. An effective optimization algorithm generates good weight vector solutions in a few optimization trial runs owing to its inherent ability to escape local minima, where a less effective algorithm requires a larger number of trial runs. Repetitive training and testing is a tedious process, so that an effective algorithm is desirable to reduce training time and increase the quality of the set of available weight-vector solutions. We present leap-frog as a robust optimization algorithm for training neural networks. In this paper the dynamic principles of leap-frog are described together with experiments to show the ability of leap-frog to generate reliable weight-vector solutions. Performance histograms are used to compare leap-frog with a variable-metric method, a conjugate-gradient method with modified restarts, and a constrained-momentum based algorithm. Results indicate that leap-frog performs better in terms of classification error than the remaining three algorithms on two distinctly different test problems. PMID- 10372760 TI - Sparsification from dilute connectivity in a neural network model of memory. AB - Several hypotheses concerning implementations of associative memory in the brain rely on analyses of the capabilities of simple network models. However, the low connectivity of cerebral networks imposes constraints which sometimes do not arise clearly from such analyses. We investigate an aspect of a simple, dilute network's operation that is sometimes overlooked, namely the setting of activation thresholds. An examination of several criteria for optimal threshold assignment affords several new insights. It becomes apparent that the network's capacity (which is simply derived) is insufficient to characterize the quality of its performance. We derive the degree of 'sparsification' or decrease in firing probability that arises from dilution, and also the consequent losses in representational ability, and propose that they should also be taken into account. To evaluate the model's performance and suitability, we argue that one should explicitly consider the trade-off that exists between storage of patterns and preservation of information, and its consequent constraints. PMID- 10372761 TI - Do cortical maps adapt to optimize information density? AB - Topographic maps are found in many biological and artificial neural systems. In biological systems, some parts of these can form a significantly expanded representation of their sensory input, such as the representation of the fovea in the visual cortex. We propose that a cortical feature map should be organized to optimize the efficiency of information transmission through it. This leads to a principle of uniform cortical information density across the map as the desired optimum. An expanded representation in the cortex for a particular sensory area (i.e. a high magnification factor) means that a greater information density is concentrated in that sensory area, leading to finer discrimination thresholds. Improvement may ultimately be limited by the construction of the sensors themselves. This approach gives a good fit to threshold versus cortical area data of Recanzone et al on owl monkeys trained on a tactile frequency-discrimination task. PMID- 10372763 TI - Alpha rhythm emerges from large-scale networks of realistically coupled multicompartmental model cortical neurons. AB - Conical pyramidal and stellate neurons were simulated using the GENESIS simulation package. Model neurons were leaky integrate-and-fire and consisted of from four to nine passive compartments. Neurophysiological measurements, based on single-cell recordings and patch-clamp experiments, provided estimations for the simulation of cortical neurons: transmitter-activated conductances, passive membrane time constants and axonal delays. Network connectivity was generated using a previously described probabilistic scheme based on known cortical histology, in which the probability of connections forming between one neuron and another fell off monotonically with increasing inter-cellular separation. Simulations of up to 6400 cortical neurons, approaching the scale of an individual cortical column, confirmed previous findings with smaller networks. Limit-cycle behaviour emerged in the network, in the frequency in the range of the mammalian alpha and beta rhythms (8-20 Hz). Contrary to expectation, near linear relationships were found between the mean soma membrane potential and and neuronal firing probability. Some of the implications for cortical information processing, in particular the dynamical interactions between the neuronal and larger scales, are discussed. PMID- 10372762 TI - Computational differences between asymmetrical and symmetrical networks. AB - Symmetrically connected recurrent networks have recently been used as models of a host of neural computations. However, biological neural networks have asymmetrical connections, at the very least because of the separation between excitatory and inhibitory neurons in the brain. We study characteristic differences between asymmetrical networks and their symmetrical counterparts in cases for which they act as selective amplifiers for particular classes of input patterns. We show that the dramatically different dynamical behaviours to which they have access, often make the asymmetrical networks computationally superior. We illustrate our results in networks that selectively amplify oriented bars and smooth contours in visual inputs. PMID- 10372764 TI - Model-independent mean-field theory as a local method for approximate propagation of information. AB - We present a systematic approach to mean-field theory (MFT) in a general probabilistic setting without assuming a particular model. The mean-field equations derived here may serve as a local, and thus very simple, method for approximate inference in probabilistic models such as Boltzmann machines or Bayesian networks. Our approach is 'model-independent' in the sense that we do not assume a particular type of dependences; in a Bayesian network, for example, we allow arbitrary tables to specify conditional dependences. In general, there are multiple solutions to the mean-field equations. We show that improved estimates can be obtained by forming a weighted mixture of the multiple mean field solutions. Simple approximate expressions for the mixture weights are given. The general formalism derived so far is evaluated for the special case of Bayesian networks. The benefits of taking into account multiple solutions are demonstrated by using MFT for inference in a small and in a very large Bayesian network. The results are compared with the exact results. PMID- 10372765 TI - The Mailbase hand surgery electronic mailing list. AB - Hand - an Internet based electronic mailing list - enables surgeons with a common interest in hand surgery to share experiences and engage in worldwide electronic scholarly discussion. This paper examines the thinking behind the setting up of this list and gives examples of how the list is being used. PMID- 10372766 TI - The Mantero flexor tendon repair in zone 1. AB - Mantero and colleagues have reported a modification of the Bunnell pull-out method for the repair of zone 1 flexor digitorum profundus (FDP) lacerations that allows active postoperative mobilization. We report a series of 24 FDP lesions in 20 adult patients treated with this technique. The mean duration of the rehabilitation regimen, which was followed by all patients, was 4.2 months. Functional assessment using Strickland's criteria demonstrated 23 excellent to good results and one poor due to a septic rupture. Nineteen of the 20 patients were satisfied with treatment and all but one of the patients returned to work within an average of 2.6 months after operation. In comparison to other zone 1 repair methods with active mobilization regimens, the Mantero technique gives better functional outcomes and appears to be more reliable. PMID- 10372767 TI - Experimental study of two new flexor tendon suture techniques for postoperative early active flexion exercises. AB - We used a rabbit model to test the postoperative mechanical strengths of two new tendon suture techniques. These were compared with the conventional modified Kessler and double looped suture techniques. For each technique, maximum load until 3 mm gap, load at 1 mm gap and ultimate load were measured at the time of operation and at weeks 1 and 3 after operation. Maximum load until 3 mm gap and load at 1 mm gap were significantly higher in the new techniques than in the conventional techniques at the time of operation and at 1 week; there was no statistical difference between the four techniques at 3 weeks. No technique resulted in a decrease in maximum load until 3 mm gap, load at 1 mm gap and ultimate load at 1 week. The new techniques reported here have the potential to withstand early active flexion exercises. PMID- 10372768 TI - Long term outcome of early active mobilization following flexor tendon repair in zone 2. AB - We reviewed 34 patients with 65 zone 2 flexor tendon injuries in 39 digits whose outcome had been prospectively studied in an earlier investigation at a mean of 10.6 years after repair. Grip strength was assessed using a Jamar dynamometer. Outcome of grip strength in relation to normal data indicated an excellent or good outcome in 94% of patients, fair in 3% and poor in 3%. Using the grading system recommended by the American Society for Surgery of the Hand, the active range of motion was graded as excellent or good in 75% of digits, fair in 15% and poor in 10%. These results compare favourably with those of the original study with 5/39 of digits showing an improvement from good to excellent. Sixteen of the 34 of patients continued to suffer from cold sensitivity. PMID- 10372769 TI - Microsurgical reconstruction of partial thumb defects. AB - We have reconstructed thumb defects using microsurgical techniques in 43 patients. The flap survival was 100% and functional improvement with near normal appearance was obtained in the reconstructed thumbs. In order to obtain satisfactory results, donor sites were confined to the great toe and its adjacent structures and adventitia was removed from the vascular pedicle, which was transferred by subcutaneous tunnelling to minimize scar formation in the reconstructed thumb. The width of transferred nail and pulp volume were matched to the defect in the thumb before transfer. PMID- 10372770 TI - Microscopic anatomy of the posterior interosseous and median nerves at sites of potential entrapment in the forearm. AB - We describe the macroscopic and microscopic anatomy of the posterior interosseous (PIN) and median nerves, at the arcade of Frohse and pronator arch respectively, in nerves obtained from five cadavers. Nerves were either constricted at sites of potential entrapment; appeared swollen proximal to these sites; or exhibited neither swelling nor constriction. Renaut bodies were present in all nerves. In the PIN, most Renaut bodies were found beneath a tendinous arcade of Frohse, whereas in the median nerve most were found proximal to the pronator arch. We propose that since Renaut bodies appear to displace normal endoneurial components, and may be associated with low-grade axonal drop-out, their presence may adversely affect the functional outcome of surgical decompression of either the PIN or median nerve. PMID- 10372771 TI - Wrist extensor muscle pathology in lateral epicondylitis. AB - The morphology of the extensor carpi radialis brevis (ECRB) muscle was investigated in 20 patients with longstanding lateral epicondylitis. Muscle biopsies were obtained from the proximal or distal portion of the ECRB and analysed by enzyme- and immunohistochemical methods. Morphological abnormalities were significantly more frequent in patients than controls and included moth eaten fibres, fibre necrosis and signs of muscle fibre regeneration as well as higher percentages of the fast-twitch oxidative (type 2A) fibre type. Changes were equally distributed proximally and distally. It is concluded that these changes, directly or indirectly, may reflect the cumulative effect of mechanical and/or metabolic overload and that decreased muscular performance in patients with lateral epicondylitis may be due to both elbow pain and physical damage to the ECRB muscle. PMID- 10372772 TI - The hand injury severity scoring system and workers' compensation cases in Wisconsin, USA. AB - The Hand Injury Severity Score was retrospectively applied to a group of workers' compensation cases in Wisconsin, USA. A statistically significant correlation was found between the score and the time interval between injury and the end of healing. These results are comparable to the findings in the original study of Campbell and Kay (1996). We provide some suggestions for further development of this scoring system. PMID- 10372773 TI - Time off work after occupational hand injuries. AB - This study analysed the impact of several factors on the start and duration of time off work among 802 patients with occupational hand injuries, in order to identify prognostic indicators. The study showed that external factors such as work and social condition seemed to have less influence on time off work than expected, whereas advice from doctors, flashbacks and impairment symptoms were important determinants. PMID- 10372774 TI - Hand injuries in agricultural accidents. AB - The aim of this study was to investigate the incidence of hand injuries due to farming accidents in a defined population with a representative mixture of agricultural activities. During a 12-month period all agricultural accidents treated at the five hospitals in the County of Ringkobing, Denmark were prospectively registered. Follow-up was done by telephone interview 4 months after the accident. Of the 260 persons injured in agricultural accidents, 117 (45%) had lesions of the upper extremity and 73 persons (28%) had hand injuries. The most common injuries were lacerations and amputations (45%) followed by fractures (36%). Mean sick leave was 25 days, and mean work impairment was 31 days in patients with hand injuries. PMID- 10372775 TI - Time off work due to scaphoid fractures and other carpal injuries in The Netherlands in the period 1990 to 1993. AB - This study assessed the epidemiology, treatment, disability and time off work due to carpal injuries in the Netherlands in the period from 1990 to 1993. Most injuries were scaphoid fractures and carpal instabilities were rare. The time off work was considerable (mean, 155 days; median, 105 days; range, 12-1708 days). Patients with non-scaphoid fractures had the shortest time off work, followed by those with scaphoid fractures; patients with carpal instabilities had the longest time off work. Despite the significant time off work, the prognosis for return to work was excellent. PMID- 10372776 TI - Algodystrophy and its association with Dupuytren's disease. AB - Seventy-two patients were examined 9 weeks after sustaining a Colles' fracture of the wrist for evidence of algodystrophy. They were examined 18 months later for evidence of Dupuytren's disease to determine the incidence of the association between the two conditions. Forty-one per cent of all patients had evidence of Dupuytren's disease at 18 months following Colles' fracture. Sixty-seven per cent of patients with algodystrophy had evidence of Dupuytren's disease compared with 19% of patients who showed no features of algodystrophy. PMID- 10372777 TI - Neurophysiological findings in vibration-exposed male workers. AB - Fractionated nerve conduction, vibrotactile sense, and temperature thresholds were studied in 73 symptomatic vibration-exposed male workers. Three symptomatic groups were distinguished: patients with isolated sensorineural symptoms; with isolated vasospastic problems; and with both. Clinical carpal tunnel syndrome occurred in 14 patients and abnormal cold intolerance (without blanching of the fingers) in 23. In the group as a whole, nerve conduction studies were abnormal in the median nerve but not in the ulnar nerve and vibration perception and temperature thresholds were impaired. Of the three symptomatic groups, patients with isolated sensorineural symptoms differed from controls. No differences were seen between patients with and without clinical carpal tunnel syndrome. With severe sensorineural symptoms the vibration perception thresholds, but not the values of the nerve conduction studies, were further impaired. The results indicated two injuries that are easily confused: one at receptor level in the fingertips and one in the carpal tunnel. Careful clinical assessment, neurophysiological testing, and examination of vibrotactile sense are required before carpal tunnel release should be considered in these patients. PMID- 10372778 TI - Dynamic traction for unstable fractures of the distal radius. AB - We have designed a flexible distracter through which dynamic traction is applied to fractures of the distal radius. The distraction is maintained during flexion and extension as well as radial and ulnar deviation. The results in 30 patients showed that the majority of the fractures maintained reduction during the period of dynamic traction. In several patients the radiological variables even improved during dynamic traction. The median radial shortening was 0 mm, the palmar angulation 7 degrees, and the radial inclination 23 degrees. Continuous dynamic traction applied to fractures of the distal radius did not lead to redisplacement of the fracture, and the overall functional results were good. PMID- 10372779 TI - Blatt's capsulodesis for chronic scapholunate dissociation. AB - We have reviewed prospectively 44 cases of chronic scapholunate dissociation treated by Blatt's dorsal capsulodesis. The diagnosis was based on clinical and arthroscopic criteria. The minimum follow-up was 2 years. The results were analysed clinically and radiologically. Postoperatively statistically significant reductions in wrist movements and grip strengths were noted. Delay in surgery and presence of compensation claims were also statistically significant factors. Patients with a high column/row index had higher overall good and excellent results. The scapholunate gap, scapholunate angle, carpal height and the type of instability as diagnosed on arthroscopy and cineradiography did not affect the outcome significantly. The scapholunate gap, scapholunate angle and the carpal height did not change significantly after operation. PMID- 10372780 TI - Late treatment of unreduced perilunate dislocations. AB - Twenty-eight patients with perilunate dislocations that had been untreated for a minimum of 6 weeks after injury were assessed at a mean of 6.8 years after subsequent treatment. Treatment consisted of open reduction with or without internal fixation of the scaphoid in six patients, proximal row carpectomy in 16, total excision of the lunate in four, and carpal tunnel release and partial excision of the lunate in two. Open reduction yielded satisfactory results in cases of less than 2 months standing. We believe that proximal row carpectomy should be considered in the treatment of chronic perilunate dislocations in patients who are seen later than 2 months after injury, if the cartilage of the proximal pole of the capitate is well preserved. The results of lunate excision were less favourable. PMID- 10372781 TI - Neuromuscular electrical stimulation and dynamic bracing as a treatment for upper extremity spasticity in children with cerebral palsy. AB - We have investigated a therapeutic regimen using neuromuscular electrical stimulation (NMES) and dynamic bracing to assess their effectiveness in reducing upper-extremity spasticity in children with cerebral palsy. Nineteen patients between 4 and 21 years of age with documented diagnoses of spastic cerebral palsy were treated. The patients included in the study followed a regimen of two 30 minute sessions of NMES of the antagonist extensors combined with dynamic orthotic traction during the day. A static brace was used at night. Spasticity of the wrist and fingers was assessed periodically using the Zancolli classification. Treatment ranged from 3 to 43 months. After treatment with electrical stimulation and dynamic bracing, all the patients moved up 1 to 3 levels in the Zancolli classification and showed a marked improvement in upper extremity function. These results show that combining NMES and dynamic orthotic traction dramatically decreases spasticity of the upper extremity in young patients with cerebral palsy. PMID- 10372782 TI - Extensor tendon dislocation in cerebral palsy. AB - An 18-year-old man with cerebral palsy presented with a flexion deformity of the middle finger particularly at the metacarpophalangeal joint and ulnar dislocation of the extensor tendon. Releasing the tight ulnar sagittal band and imbricating the attenuated radial sagittal band allowed centralization of the extensor tendon. For complete correction of other deformities intrinsic release and extrinsic flexor muscle lengthening were done. Extensor tendon instability in this case was due to the combined forces of the extrinsic and intrinsic muscles on the retinacular system of the extensor mechanism. PMID- 10372783 TI - Biceps-to-triceps transfer in tetraplegia. The medial route. AB - Eight tetraplegic patients (13 elbows) were treated by biceps-to-triceps transfer. To avoid the risk of radial nerve injury, we chose a medial routing of the biceps. The mean follow-up after surgery was 17.8 months (range, 4-47 months). No complications were encountered. Active extension of the elbow was achieved in each case. The mean postoperative active range of motion of the elbow was 6 degrees extension and 137 degrees flexion. After the biceps-to-triceps transfer mean extension torque of the elbow was 3.7 Nm and mean flexion torque was 10 Nm. In eight elbows in which it was measured, there was a 47% reduction in elbow flexion power. Nevertheless no patient complained about that reduction, and all of them were satisfied. PMID- 10372784 TI - The bilobed flap in treatment of mucous cysts of the distal interphalangeal joint. AB - The bilobed flap has many uses in the field of plastic and reconstructive surgery. We describe its use in achieving skin cover following excision of mucous cysts in six digits in six patients, with a minimum follow up of 1 year. There were no postoperative complications. No cyst has recurred and cosmesis has been excellent in all cases. PMID- 10372785 TI - Use of the homodigital adipofascial turnover flap for dorsal cover of distal interphalangeal joint defects. AB - The homodigital adipofascial turnover flap was originally described by Voche and Merle (1994) for dorsal cover of the proximal interphalangeal joint. We present three patients in whom this flap was used to cover dorsal defects of the distal interphalangeal joint, and describe an adaptation to allow greater flap mobility. PMID- 10372786 TI - In vivo MR studies of dynamic changes in the interosseous membrane of the forearm during rotation. AB - In vivo dynamic changes in the interosseous membrane (IOM) during forearm rotation were studied using magnetic resonance imaging (MRI). The right forearms of 20 healthy volunteers were examined in five different rotational positions. Axial slices were obtained at the proximal quarter, the middle and the distal quarter of the forearm. The changes in shape of the IOM during rotation were observed in an axial MR plane. For each image, we measured the interosseous distance and the length of the interosseous membrane. Images of the tendinous and membranous parts of the IOM could be differentiated by thickness. There were minimal dynamic changes in the tendinous part on the MRI while the membranous part showed numerous changes during rotation. The interosseous distance and the length of the interosseous membrane were maximum from a neutral to a slightly supinated position. The tendinous part is considered to be taut during rotation to provide stability between the radius and the ulna, but the membranous part which is soft, thin and elastic, allows smooth rotation. PMID- 10372787 TI - Endoscopic release of the carpal tunnel: a technical note. AB - The Agee technique is a standard approach for the endoscopic release of the carpal tunnel. We describe a modification of the technique. This involves dilatation of the path for the blade assembly with urethral dilators. This permits a graded dilatation of the path. We believe it creates a track of more uniform width. It also provides more accurate information about the tightness of the carpal tunnel. Its adoption may reduce the chance of nerve injury. PMID- 10372788 TI - An aid to digit replantation: a new use for the Harris silicone tile. AB - We have used the Harris silicone tile to provide secure anchorage of amputated digits during dissection and preparation for replantation. Sound anchorage is obtained and the identification and dissection of structures is facilitated. PMID- 10372789 TI - A reliable technique for radiographic imaging of the pisotriquetral joint. AB - We report a novel technique for radiographic imaging of the pisotriquetral joint. This technique has proved useful for five consecutive patients presenting with pisotriquetral osteoarthritis. It is consistently reliable in imaging the joint and reduces the number of unsuccessful radiographic attempts. PMID- 10372790 TI - Triplication of the thumb. AB - A child with triplication of the thumb is presented. Each thumb was fully developed with neurovascular bundles, flexor and extensor tendons. Although all three thumbs were triphalangeal, they shared a common metacarpal and two shared a common proximal phalanx. Since this anomaly does not fit in the existing accepted Wassel (1969) classification, a new category is suggested. PMID- 10372791 TI - Bilateral proximal radial and scaphoid fractures in a child. AB - A 13-year-old boy fell and suffered concomitant bilateral fractures of the proximal radius and scaphoid. Ipsilateral fractures of the proximal radius and scaphoid have been reported only once previously in a child, and never bilaterally. This article reviews paediatric proximal radial fractures and scaphoid fractures and their associated injuries. This report underlines the importance of examining for other injuries when a child presents with an apparently isolated upper extremity fracture. PMID- 10372792 TI - Behavioral profile of constituents in ayahuasca, an Amazonian psychoactive plant mixture. AB - Ayahuasca is a psychoactive plant mixture typically composed of the beta carboline-rich Banisteriopsis caapi vine and the hallucinogenic plant Psychotria viridis. Ayahuasca has long been used by aboriginal populations for its putative spiritual and medicinal benefits. Although the presumed primary chemical constituents of ayahuasca have been identified, little is known about the basic in vivo pharmacology of the extract. Two principal constituents of ayahuasca, the beta-carboline harmine and N,N-dimethyltryptamine (DMT) were selected for detailed study in mice using the Functional Observational Battery (FOB). The B. caapi extract was then examined alone and in combination with DMT. Harmine and the B. caapi extract produced similar effects in the FOB, particularly in the open field. Clonic and tonic motor movements were augmented by DMT administration. Harmine and B. caapi decreased acoustic startle amplitude without significantly affecting prepulse inhibition. DMT appeared to attenuate startle decreasing effects of harmine and B. caapi, although these effects fell just short of significance. These results suggest that the behavioral effects of B. caapi in mice may be attributed in large part to its principal alkaloid species, harmine, and related beta-carbolines in the extract. Hence, the presence of the banisteriopsis vine in the admixture may directly contribute to the unique subjective effects of ayahuasca. PMID- 10372793 TI - Relationship of ADHD, depression, and non-tobacco substance use disorders to nicotine dependence in substance-dependent delinquents. AB - This study used standardized interviews to examine the relationship of attention deficit hyperactivity disorder (ADHD), major depression (MDD), and other illicit substance use disorders (SUD) to onset and severity of nicotine dependence in 82 female and 285 male adolescents with conduct disorder (CD) and SUD. Results indicate that both ADHD and MDD significantly contribute to severity of nicotine dependence in delinquents with SUD. ADHD is further associated with earlier onset of regular smoking in males. Severity of non-tobacco SUD also was related directly to nicotine dependence severity in both males and females, and to earlier onset of smoking in males. Our findings illuminate the contribution of comorbidity to nicotine dependence and its relationship to other SUD severity among adolescents with CD and SUD and highlight the need for coordinated assessment and treatment of smoking cessation along with concurrent treatment of other drug use and psychiatric comorbidity such as ADHD and MDD in such youths. PMID- 10372794 TI - Enhanced treatment outcomes for cocaine-using methadone patients. AB - Cocaine dependent methadone patients were randomly assigned to 6 months of high intensity cognitive-behavioral therapy or low intensity therapy. A repeated measures ANOVA was conducted with patients stratified on severity of cocaine use at baseline. Both treatment groups showed significant and equivalent reductions in cocaine use during the post-treatment period. Completing either therapy and lower cocaine severity at baseline were associated with lower proportion of cocaine-positive urines across a 48-week post-treatment period. Examination of the treatment x cocaine severity interaction provided some evidence that high severity patients improved more if exposed to high intensity treatment than to low intensity treatment. Positive outcomes for therapy completers relative to non completers increased over time. The results are consistent with several clinical trials showing that: (1) participation in treatment is associated with reductions in cocaine use; and (2) the relationship between treatment intensity and outcome is not linear and may better be explained by an interaction between patient and treatment factors. PMID- 10372795 TI - Alternative definitions of high risk for impaired driving: the overlap of high volume, frequent heavy drinking and alcohol dependence. AB - This paper examines the distributions of past-year volume of ethanol intake, frequency of drinking 5+ drinks and alcohol dependence in a representative sample of 18,352 U.S. current drinkers aged 18 years or over. Within categories defined by these three partially overlapping domains, it presents rates of self-perceived impaired driving, i.e. driving after having had too much to drink, in the year preceding interview. High volume drinkers, those with an average daily ethanol intake of 1 ounce or more, composed 19.7% of current drinkers and accounted for 66.5% of all reported ethanol consumption, 72.6% of all heavy drinking days, 49.2% of all alcohol dependence and 62.8% of all impaired driving incidents. Frequent heavy drinkers, those who drank 5+ drinks at least once a week, composed 12.3% of current drinkers and accounted for 42.9% of all reported ethanol consumption, 81.9% of all heavy drinking days, 40% of all alcohol dependence and 57% of all impaired driving incidents. Drinkers with DSM-IV alcohol dependence composed 9.9% of current drinkers and accounted for 28.9% of all reported ethanol consumption, 37% of all heavy drinking days and 56.9% of all impaired driving incidents. The overlap of these three high risk groups, each of which had a probability of at least one impaired driving incident per year, was far from complete. Of individuals who met any of these criteria for high risk drinking (i.e. high volume, frequent heavy drinking or dependence), more than half met only one criterion and only one in seven met all three. The group that did meet all three criteria had such a high rate of impaired driving incidents, an average of 5.14/year, that it accounted for 36.4% of all such incidents despite making up only 3.8% of all current drinkers. The results are discussed in terms of their implications for targeting prevention and intervention efforts, e.g. whether targeting one problematic aspect of drinking behavior will reach drinkers with other types of problem behaviors as well. PMID- 10372796 TI - Marijuana initiates and their impact on future drug abuse treatment need. AB - This paper presents estimates of the number of people who will need treatment for illicit drug abuse problems for the years 2000 through 2020. The methodology employs logistic regression models, with treatment need as a dependent variable, using data from lifetime marijuana users included in the National Household Survey on Drug Abuse. Age at first use of marijuana was found to be the most important predictor in these models. Other variables included in the models were age, gender, and race/ethnicity. By generating estimates under alternative assumptions about future rates of initiation, it was projected that if current rates of initiation continue, treatment need will increase by 57% by 2020, and that the need for treatment will remain high even if initiation rates decrease dramatically, because of the aging baby boom cohort. PMID- 10372798 TI - Beliefs about smoking cessation among out-of-school youth. AB - Although the majority of adolescents in the 13-18 age range are at school, there is a need to target specific groups of young smokers such as unemployed youth. For those young people who are not at school, few directed programs are available in either prevention or cessation and information is needed about the design and delivery of appropriate programs for this population. This report presents the results from a survey of unemployed youth and students at vocational colleges about various aspects of smoking cessation. The majority of out-of-school youth smokers had not tried to quit, but 52% were contemplating action to quit. Only a quarter of the smokers had quit for more than a week. Few young smokers would use a recognised program though more females would change to a lower nicotine brand, quit with the help of a friend or participate in a group quit program. The method of quitting most would recommend to peers is 'use of will power'. Incentives to quit were attractive to only a third of the smokers, and many enhancing and inhibiting factors for participation in programs were identified. In particular, efforts to quit increased their confidence in quitting, supporting the need to assist those who are contemplating action to quit. Programs need to incorporate input from youth and be tailored for them but not necessarily for different groups such as non-secondary school students and unemployed youth. PMID- 10372797 TI - Inhibitors of cytochrome P450 differentially modify discriminative-stimulus and antinociceptive effects of hydrocodone and hydromorphone in rhesus monkeys. AB - The present study was conducted to investigate the role of cytochrome P450 in the discriminative-stimulus and antinociceptive effects of hydrocodone (HC) and hydromorphone (HM) in rhesus monkeys. In morphine-deprived monkeys, morphine dose dependently reversed naltrexone-lever responding, an effect also produced by HC and HM. HC and HM also produced antinociception in a warm-water tail withdrawal procedure. Budipine and naltrexone shifted the dose-effect curves for the discriminative-stimulus effects of HC and HM to the right. In contrast, naltrexone, but not budipine (10.0 mg/kg) or quinidine (10.0 mg/kg), dose dependently antagonized the antinociceptive effects of HC. Budipine and quinidine decreased the concentration of HM in plasma without significantly affecting the levels of HC, suggesting that these CYP2D6 inhibitors decreased the conversion of HC HM. Thus, some behavioral effects of HC are not modified by a marked inhibition of CYP2D6, suggesting that these effects of HC are not due to its conversion to HM but, rather, that both HC and HM act directly on mu receptors. PMID- 10372799 TI - Alcohol abusers who want to quit smoking: implications for clinical treatment. AB - Although most alcohol abusers are dependent on nicotine, studies of such individuals have been scarce. Consequently, little information is available for advising clients who wish to consider resolving both problems. Clients entering an outpatient alcohol treatment program who were also current smokers were asked about their temporal preferences for changing their alcohol and cigarette use. Over three-quarters of alcohol abusers who were also smokers when asked said they would be willing to consider stopping smoking during or after treatment for an alcohol problem. Individuals who were interested in quitting smoking cigarettes while in treatment for an alcohol problem were different from those who did not want to stop smoking, and such differences may influence their ability to successfully address both problems together. Compared to those who preferred to change their drinking first then address their smoking, those who said they would be willing to address both problems (i.e. smoking and drinking) together in treatment were not only considerably more likely to think that quitting smoking would affect quitting drinking, but also more likely to be planning to quit smoking in the next six months. These results suggest that some individuals whose alcohol problems are not severe and who also smoke cigarettes will be more receptive to a dual recovery approach than others. In the absence of research findings, health care practitioners who encounter individuals who drink heavily and smoke cigarettes should at a minimum explore the option of dual cessation with their clients. The clinical and research implications of the present results are discussed. PMID- 10372800 TI - Regulatory role of SH3 domain-mediated protein-protein interactions in synaptic vesicle endocytosis. AB - Src homology (SH) 3 domains are small modules found in a diverse array of proteins. The presence of an SH3 domain confers upon its resident protein the ability to interact with specific proline-rich sequences in protein binding partners. A major focus of research has highlighted a role for SH3 domain mediated interactions in the regulation of signal transduction events. However, more recent data has suggested an important function for SH3 domains in vesicular trafficking. This review will focus on this newly emerging role with a particular emphasis on the molecular components involved in synaptic vesicle endocytosis and the regulatory role of SH3 domain-mediated protein-protein interactions in this process. PMID- 10372801 TI - Mechanisms of nitric oxide-dependent apoptosis: involvement of mitochondrial mediators. AB - Programmed cell death occurs in several physiopathological situations in multicellular organisms and constitutes a common mechanism of cell replacement, tissue remodelling and removal of altered cells. The effectors that induce apoptosis as well as the signalling pathways involved in the process are the subjects of current work. In addition to receptor-mediated apoptosis, highly reactive molecules, such as NO, influence cell viability either by acting as a protection against apoptogenic stimuli, or by inducing apoptosis when produced at elevated concentrations. The contribution to apoptosis of mediators released by the mitochondria and involved in the activation of caspases focused attention on the functional changes caused by NO in this organelle. NO induces mitochondrial permeability transition and promotes apoptosis in cell-free systems containing mitochondria and nuclei. Moreover, NO-dependent apoptosis can be blocked in most cases through the use of permeability transition or caspase inhibitors. The intracellular pathways activated in response to NO challenge and involved in the regulation of apoptosis are analysed. PMID- 10372802 TI - Intensification of growth factor receptor signalling by phorbol treatment of ligand-primed cells implies a dimer-stabilizing effect of protein kinase C dependent juxtamembrane domain phosphorylation. AB - Protein kinase C (PKC) phosphorylates the juxtamembrane domain of many growth factor receptors, but the physiologic effect of this modification on ligand signalling and desensitisation is unclear. Here we show that PKC-dependent transmodulation of EGFR and ErbB2 signalling is schedule-specific: prolonged pre treatment of A431 cells with the PKC agonist phorbol dibutyrate potently inhibits subsequent ligand-induced EGFR signalling as expected, but EGF pre-treatment reverses the inhibitory effect of phorbol. The agonist activity of PKC on receptor signalling is even more apparent when cells are treated with phorbol in the presence of a tyrosine phosphatase inhibitor. Because these findings suggested a synergistic interaction between tyrosine- and PKC-dependent phosphorylation events, we sought to define the interactions of tyrosine phosphorylated and PKC-modified ErbB2 subsets within EGF-inducible hetero oligomers. Growth factor-dependent PKC transphosphorylation takes place exclusively within endocytosed tyrosine-phosphorylated receptor oligomers. Moreover, phorbol differentially affects two ErbB2 C-terminal autophosphorylation sites: whereas phosphorylation of Tyr1222 is reduced, phosphorylation of Tyr1139 is increased. These results suggest that PKC-dependent phosphorylation of the juxtamembrane domain may contribute positively to both internalisation and signalling of ligand-activated receptors, simultaneously accelerating termination of growth factor action. We propose that transient PKC-dependent signal amplification results from enhanced stability of liganded receptor oligomers due to phosphorylation-dependent juxtamembrane domain interactions, analogous to the protein-protein binding now known to be induced by serine-threonine phosphorylation of CREB and SMAD. PMID- 10372803 TI - Requirement of multiple SH3 domains of Nck for ligand binding. AB - The Nck adaptor protein comprises a single C-terminal SH2 domain and three SH3 domains. The domain structure of Nck suggests that Nck links tyrosine kinase substrates to proteins containing proline-rich motifs. Here we show that Bcr/Abl tyrosine kinase, and three tyrosine phosphorylated proteins (115, 120 and 155 kDa) are co-immunoprecipitated with antibody against Nck from lysates of the human leukaemia cell line K562. By means of affinity purification with the Nck binding phosphopeptide EPGPY(P)AQPSV, we could also detect the association of endogenous Nck with the proto-oncogene product Cbl. An investigation of the nature of interactions revealed that Bcr/Abl, Cbl, and the 155-kDa tyrosine phosphotyrosine bind exclusively to the SH3 domains of Nck. In addition, none of the single SH3 domains of Nck expressed as glutathione-S-transferase (GST) fusion proteins is able to interact with the proline-rich ligands. However, combined first and second SH3 domains have the capacity to bind Bcr/Abl, Chl and p155. Mutations of conserved tryptophan to Lysine in either of the combined first and second SH3 domains completely abolish ligand binding. These data suggest that cooperation exists among the SH3 domains of Nck for a high-affinity binding of proteins containing proline-rich motifs. PMID- 10372804 TI - Involvement of intermediary metabolites in the pathway of extracellular Ca2+ induced mitogen-activated protein kinase activation in human fibroblasts. AB - Human fibroblasts in culture will grow in serum-free medium containing serum replacement factors, but without protein growth factors, as long as the Ca2+ level is 1.0-2.0 mM. When the Ca2+ is reduced to 0.1 mM, the cells stop cycling, but they can be reinduced to cycle by raising the Ca2+ level to 1.0 mM Ca2+ or to higher concentrations that result in activation of mitogen-activated protein kinase (MAPK). We now report that exposure of human fibroblasts to extracellular Ca2+ increased the level of inositol (1,4,5)-trisphosphate in the cytoplasm and caused a transient rise in the concentration of intracellular free Ca2+. Ca2+ induced MAPK activation was partly abolished by treatment of the cells with pertussis toxin. It was also decreased by treatment of cells with thapsigargin, which depletes intracellular Ca2+ stores; with phorbol 12-myristyl 13-acetate (PMA), which down-regulates protein kinase C (PKC); with the calmodulin antagonists N-(6-aminohexyl)-5-chloro-1-naphthalenesulphonamide HCl (W-7), and calmidazolium (24571); as well as with lanthanum, a Ca2+ channel inhibitor. Ca2+ stimulation did not result in phosphorylation of the c-raf-1 protein. Our results suggest that extracellular Ca2+ stimulates MAPK activation through a pathway(s) involving a pertussis toxin-sensitive G protein, phospholipase C, intracellular free Ca2+, calmodulin, and PKC. PMID- 10372805 TI - Signalling events mediating the activation of protein kinase C by interleukin-2 in cytotoxic T cells. AB - Interleukin-2 (IL-2) plays a vital role in the generation and regulation of the immune response, including important aspects of T cell survival. IL-2-mediated survival of T cells appears to be dependent on the activation of a pool of membrane-associated protein kinase C (PKC) that occurs in the absence of detectable translocation of the enzyme from the cytosol to membranes. In this report we investigate the mechanism(s) responsible for this PKC activation after IL-2 stimulation in the cytotoxic T cell line, CTLL-2. Tyrosine kinase activity, activated after IL-2 stimulation, was found not to be linked to the activation of PKC by the cytokine. On the other hand, a pertussis toxin (PTX)-sensitive G protein did appear coupled to PKC activation since PTX effectively blocked IL-2 stimulated PKC activity. Diacylglycerols (DAG), but not inositol 1,3,5 triphosphate (IP3) and intracellular Ca2+, increased after IL-2 stimulation suggesting that DAGs were generated via the phosphatidylcholine-phospholipase C (PC-PLC) or phosphatidylcholine-phospholipase D (PC-PLD) pathways. The increase in DAG by IL-2 was probably necessary for activation of membrane-resident PKC since exogenously applied DAG stimulated this PKC pool in both intact cells and in isolated membranes. IL-2 also increased arachidonic acid (AA) production in CTLL-2 cells, probably via phospholipase A2 (PLA2) since the PLA2 inhibitors oleoyloxyethyl phosphocholine and AACOCF3 (AACF) effectively blocked IL-2 stimulated PKC activation. Exogenous AA also increased PKC activity in intact cells and isolated membranes, suggesting that AA produced by IL-2 receptor stimulation was probably linked to PKC activation. These results suggest that the activation of membrane-resident PKC by IL-2 involves multiple second messengers, including G proteins, DAG and AA. PMID- 10372806 TI - An antigen receptor (NCCRP-1) on nonspecific cytotoxic cells is a phosphoprotein associated with the JAK-STAT activation pathway. AB - The antigen receptor (nonspecific cytotoxic cell receptor protein-1/NCCRP-1) on nonspecific cytotoxic cells (NCC) is a 32-kDa predicted Type III membrane protein. The N-terminal cytoplasmic portion of this receptor contains full length and truncated BOX-1 motifs. These motifs are also found on cytokine, erythropoietin and growth hormone receptors and provide docking sites for JAK kinases. In the present study, we investigated a relationship between NCCRP-1 and JAK2 kinase binding. A possible association with further downstream STAT activation was suggested. NCCRP-1 was phosphorylated on C-terminal domain serine residues. To examine the possibility that NCCRP-1 was associated with JAK kinase(s), NCC were purified and lysates were probed by Westen blotting (WB) for the presence of JAK2 kinase. Unlike their mammalian counterparts, NCC JAK2 kinase existed as a 90-95-kDa primary and a 35-40-kDa secondary breakdown product. Both mol wt. forms were significantly smaller than those reported for human JAK kinases. To determine if NCCRP-1 was physically associated with JAK2 kinase, chemical cross-linking experiments were conducted. NCC membrane preparations were treated with the chemical cross-linker DSS, solubilised and immunoprecipitated with anti-NCCRP-1 (e.g., 32 kDa) mab 5C6. WB analysis using anti-JAK2 mab and mab 5C6 demonstrated that the immunoprecipitate contained both the 32-kDa NCCRP-1 and 85-90-kDa JAK2 kinase. To examine further the possibility that STAT proteins may be associated with NCC/NCCRP-1 activation, NCC lysates were probed (WB) with various anti-STAT mabs. The strongest signal was produced by a 100-kDa STAT-6 protein. Lysates were negative for STAT-1, STAT-3 and STAT-5. These data indicate that the N-terminus of NCCRP-1 may initiate cytokine gene transcription by the JAK-STAT signalling pathway. PMID- 10372807 TI - Haloperidol effect on intracellular signals system coupled to alpha1-adrenergic receptor in rat cerebral frontal cortex. AB - The induction of intracellular signals coupled to alpha1-adrenoceptor by haloperidol, were studied in rat cerebral frontal cortex. The neuroleptic exerts a biphasic effect on nitric oxide synthase (NOS), inhibiting the enzymatic activity at low concentrations (10(-9) M), while higher concentrations (10(-5) M) increased it. Protein kinase C (PKC) and phosphoinositol turnover (PIs) were involved in these actions, as haloperidol induced PKC translocation at low concentrations, and increased PIs turnover at high concentrations. All the effects of haloperidol were blocked by the alpha-adrenoceptor antagonist prazosin and the phospholipase C (PLC) inhibitor NCDC. The possibility that a cross-talk between both enzymatic pathways depending on the neuroleptic concentration used in rat cerebral frontal cortex, is also discussed. PMID- 10372808 TI - Intracellular mechanisms of L-selectin induced capping. AB - Leucocyte adhesion to endothelial cells is a tightly regulated process involving selectins, integrins and immunoglobulin-like proteins. Cell adhesion and communication are controlled by membrane dynamics like receptor capping. Capping of surface receptors is an ubiquitous mechanism but still not well understood. Employing immunofluorescence techniques, we demonstrate that L-selectin triggering results in receptor capping of the L-selectin molecules in lymphocytes. Using pharmacological inhibitors and genetic deficient cell lines we show that this process involves intracellular signalling molecules. L-Selectin capping seems to be independent on activation of p56lck-kinase, but requires the neutral sphingomyelinase, small G proteins and the cytoskeleton. Therefore, capping of L-selectin upon stimulation might play an important role in the very early phase of lymphocyte trafficking. PMID- 10372809 TI - Fifth W.D.M. Paton Memorial Lecture. Waterton and Wouralia. PMID- 10372810 TI - Modification of membrane currents in mouse neuroblastoma cells following infection with rabies virus. AB - 1. The effect on membrane currents of infection of mouse neuroblastoma NA cells with rabies virus was studied by using the whole-cell patch clamp technique. 2. Three types of membrane currents, namely voltage-dependent Na+ current (I(Na)), delayed rectifier K+ current (I(K-DR)) and inward rectifier K+ current (I(K-IR)) were elicited in uninfected cells. 3. In cells 3 days after infection with the virus, no detectable change was observed in morphology and membrane capacitance, but I(Na) and I(K-IR) were significantly decreased in amplitude without any appreciable difference in the time course of current activation and inactivation. The voltage-dependence of I(Na) activation was significantly shifted in the positive direction along the voltage axis with a decreased slope. I(K-DR) remained almost unaltered after the viral infection. 4. The resting membrane potential, measured with a physiological K+ gradient across the cell membrane, was decreased (depolarized) after the viral infection. The depolarization was associated with the decreased amplitude of I(K-IR). 5. These results suggest that infection of mouse neuroblastoma NA cells with rabies virus causes reduction of functional expression of ion channels responsible for I(Na) and I(K-IR), and provide evidence for possible involvement of the change in membrane properties in the pathogenesis of rabies disease. PMID- 10372811 TI - Mechanism of verapamil block of a neuronal delayed rectifier K channel: active form of the blocker and location of its binding domain. AB - 1. The mechanism of verapamil block of the delayed rectifier K currents (I K(DR)) in chick dorsal root ganglion (DRG) neurons was investigated using the whole-cell patch clamp configuration. In particular we focused on the location of the blocking site, and the active form (neutral or charged) of verapamil using the permanently charged verapamil analogue D890. 2. Block by D890 displayed similar characteristics to that of verapamil, indicating the same state-dependent nature of block. In contrast with verapamil, D890 was effective only when applied internally, and its block was voltage dependent (136 mV/e-fold change of the on rate). Given that verapamil block is insensitive to voltage (Trequattrini et al., 1998), these observations indicate that verapamil reaches its binding site in the uncharged form, and accesses the binding domain from the cytoplasm. 3. In external K and saturating verapamil we recorded tail currents that did not decay monotonically but showed an initial increase (hook). As these currents can only be observed if verapamil unblock is significantly voltage dependent, it has been suggested (DeCoursey, 1995) that neutral drug is protonated upon binding. We tested this hypothesis by assessing the voltage dependence of the unblock rate from the hooked tail currents for verapamil and D890. 4. The voltage dependence of the off rate of D890, but not of verapamil, was well described by adopting the classical Woodhull (1973) model for ionic blockage of Na channels. The voltage dependence of verapamil off rate was consistent with a kinetic scheme where the bound drug can be protonated with rapid equilibrium, and both charged and neutral verapamil can unbind from the site, but with distinct kinetics and voltage dependencies. PMID- 10372812 TI - UK-78,282, a novel piperidine compound that potently blocks the Kv1.3 voltage gated potassium channel and inhibits human T cell activation. AB - 1. UK-78,282, a novel piperidine blocker of the T lymphocyte voltage-gated K+ channel, Kv1.3, was discovered by screening a large compound file using a high throughput 86Rb efflux assay. This compound blocks Kv1.3 with a IC50 of approximately 200 nM and 1:1 stoichiometry. A closely related compound, CP 190,325, containing a benzyl moiety in place of the benzhydryl in UK-78,282, is significantly less potent. 2 Three lines of evidence indicate that UK-78,282 inhibits Kv1.3 in a use-dependent manner by preferentially blocking and binding to the C-type inactivated state of the channel. Increasing the fraction of inactivated channels by holding the membrane potential at - 50 mV enhances the channel's sensitivity to UK-78,282. Decreasing the number of inactivated channels by exposure to approximately 160 mM external K+ decreases the sensitivity to UK 78,282. Mutations that alter the rate of C-type inactivation also change the channel's sensitivity to UK-78,282 and there is a direct correlation between tau(h) and IC50 values. 3. Competition experiments suggest that UK-78,282 binds to residues at the inner surface of the channel overlapping the site of action of verapamil. Internal tetraethylammonium and external charybdotoxin do not compete UK-78,282's action on the channel. 4. UK-78,282 displays marked selectivity for Kv1.3 over several other closely related K+ channels, the only exception being the rapidly inactivating voltage-gated K+ channel, Kv1.4. 5. UK-78,282 effectively suppresses human T-lymphocyte activation. PMID- 10372813 TI - Effects of terbutaline on force and intracellular calcium in slow-twitch skeletal muscle fibres of the rat. AB - 1. The effect of the alpha2-adrenoceptor agonist, terbutaline, was investigated on simultaneously measured force and intracellular free calcium ([Ca2+]i) in intact rat soleus muscle fibres, and on contractile protein function and Ca2+ content of the sarcoplasmic reticulum (SR) in skinned fibres. 2. Terbutaline (10 microM) had no significant effect on either resting force or [Ca2+]i. Exposure to terbutaline increased both the integral of the indo-1 ratio transient and peak twitch force by 37%. 3. At sub-maximal (10 Hz) stimulation frequencies, terbutaline accelerated force relaxation but had highly variable effects on tetanic force amplitude. The corresponding indo-1 ratio transients were significantly larger, and faster to decay than the controls. 4. Terbutaline increased tetanic force at near maximal stimulation frequencies (50 Hz) by increasing tetanic [Ca2+]i. Force relaxation was accelerated at this frequency with no significant change in the indo-1 ratio transient decay rate. 5. All of terbutaline's effects on force and indo-1 ratio transients in intact fibres were completely blocked and reversed by ICI 118551 (1 microM). 6. Mechanically skinned fibres isolated from intact muscles pre-treated with terbutaline showed no significant changes in SR Ca2+ content, myofilament [Ca2+]i-sensitivity or maximum force generating capacity. 7. The results suggest that terbutaline primarily modulates force by altering the amplitude and decay rate of the [Ca2+]i transient via phosphorylation of both the ryanodine receptor (RR) and the SR pump regulatory protein, phospholamban (PLB). The high variability of responses of slow-twitch muscles to beta2-agonists probably reflects individual differences in basal phosphorylation levels of PLB relative to that of RR. PMID- 10372814 TI - Pilocarpine modulates the cellular electrical properties of mammalian hearts by activating a cardiac M3 receptor and a K+ current. AB - 1. Pilocarpine, a muscarinic acetylcholine receptor (mAChR) agonist, is widely used for treatment of xerostomia and glaucoma. It can also cause many other cellular responses by activating different subtypes of mAChRs in different tissues. However, the potential role of pilocarpine in modulating cardiac function remained unstudied. 2. We found that pilocarpine produced concentration dependent (0.1-10 microM) decrease in sinus rhythm and action potential duration, and hyperpolarization of membrane potential in guinea-pig hearts. The effects were nearly completely reversed by 1 microM atropine or 2 nM 4DAMP methiodide (an M3-selective antagonist). 3. Patch-clamp recordings in dispersed myocytes from guinea-pig and canine atria revealed that pilocarpine induces a novel K+ current with delayed rectifying properties. The current was suppressed by low concentrations of M3-selective antagonists 4DAMP methiodide (2-10 nM), 4DAMP mustard (4-20 nM, an ackylating agent) and p-F-HHSiD (20-200 nM). Antagonists towards other subtypes (M1, M2 or M4) all failed to alter the current. 4. The affinity of pilocarpine (KD) at mAChRs derived from displacement binding of [3H] NMS in the homogenates from dog atria was 2.2 microM (65% of the total binding) and that of 4DAMP methiodide was 2.8 nM (70% of total binding), consistent with the concentration of pilocarpine needed for the current induction and for the modulation of the cardiac electrical activity and the concentration of 4DAMP to block pilocarpine effects. 5. Our data indicate, for the first time, that pilocarpine modulates the cellular electrical properties of the hearts, likely by activating a K+ current mediated by M3 receptors. PMID- 10372815 TI - Evidence for proteinase-activated receptor-2 (PAR-2)-mediated mitogenesis in coronary artery smooth muscle cells. AB - 1. This study investigates, whether in addition to the thrombin receptor (PAR-1), the proteinase-activated receptor-2 (PAR-2) is present in vascular smooth muscle cells (SMC) and mediates mitogenesis. PAR-2 is activated by low concentrations of trypsin and the synthetic peptide SLIGRL. 2. Stimulation of bovine coronary artery SMC by trypsin (2 nM) caused a 3 fold increase in DNLA-synthesis. A similar effect was observed with 10 nM thrombin. Trypsin-induced mitogenesis was inhibited by soybean trypsin inhibitor, indicating that the proteolytic activity of the enzyme was required for its mitogenic effect. 3. The specific PAR-2 activating peptide SLIGRL or the PAR1-activating peptide SFFLRN did not elicit mitogenesis. 4. When the SMC were exposed to SLIGRL (40 nM), a homologous desensitization of cytosolic Ca2+ mobilization was found after subsequent stimulation with trypsin (40 nM) but not thrombin (15 nM). 5. Trypsin (2 nM) as well as SLIGRL (100 microm) activated the nuclear factor KB (NFkappaB) with a maximum response 2 h after stimulation of the SMC. This suggests that both agonists acted via a common receptor, PAR-2. Maximum activation of NFkappaB by thrombin (10 nM) was detected after 4-5 h. 6. These data suggest that PAR-2 is present in coronary SMC and mediates a mitogenic response. Activation of NFkappaB via either PAR-1 or PAR-2 does not predict mitogenesis. PMID- 10372816 TI - Role of the medial prefrontal cortex in 5-HT1A receptor-induced inhibition of 5 HT neuronal activity in the rat. AB - 1. We examined the involvement of the frontal cortex in the 5-HT2A receptor induced inhibition of 5-HT neurones in the dorsal raphe nucleus (DRN) of the anaesthetized rat using single-unit recordings complemented by Fos immunocytochemistry. 2. Both transection of the frontal cortex as well as ablation of the medial region of the prefrontal cortex (mPFC) significantly attenuated the inhibition of 5-HT neurones induced by systemic administration of the 5-HT1A receptor agonist, 8-OH-DPAT (0.5-16 microg kg(-1), i.v.). In comparison, the response to 8-OH-DPAT was not altered by ablation of the parietal cortex. The inhibitory effect of 8-OH-DPAT was reversed by the 5-HT1A receptor antagonist, WAY 100635 (0.1 mg kg(-1), i.v.) in all neurones tested. 3. In contrast, cortical transection did not alter the sensitivity of 5-HT neurones to iontophoretic application of 8-OH-DPAT into the DRN. Similarly, cortical transection did not alter the sensitivity of 5-HT neurones to systemic administration of the selective 5-HT reuptake inhibitor, paroxetine (0.1-0.8 mg kg(-1) , i.v.). 4. 8-OH-DPAT evoked excitation of mPFC neurones at doses (0.5-32 microg kg(-1), i.v.) in the range of those which inhibited 5-HT cell firing. At higher doses (32-512 microg kg(-1), i.v.) 8-OH-DPAT inhibited mPFC neurones. 8-OH DPAT (0.1 mg kg(-1), s.c.) also induced Fos expression in the mPFC. The neuronal excitation and inhibition, as well as the Fos expression, were antagonized by WAY 100635. 5. These data add further support to the view that the inhibitory effect of 5-HT1A receptor agonists on the firing activity of DRN 5-HT neurones involves, in part, activation of a 5-HT1A receptor-mediated postsynaptic feedback loop centred on the mPFC. PMID- 10372817 TI - Anti-inflammatory activity of c(ILDV-NH(CH2)5CO), a novel, selective, cyclic peptide inhibitor of VLA-4-mediated cell adhesion. AB - 1. Small, N- to C-terminal cyclized peptides containing the leucyl-aspartyl valine (LDV) motif from fibronectin connecting segment-1 (CS-1) have been investigated for their effects on the adhesion of human T-lymphoblastic leukaemia cells (MOLT-4) to human plasma fibronectin in vitro mediated by the integrin Very Late Antigen (VLA)-4 (alpha4beta1, CD49d/CD29). 2. Cyclo(-isoleucyl-leucyl aspartyl-valyl-aminohexanoyl-) (c(ILDV-NH(CH2)5CO)) was approximately 5 fold more potent (IC50 3.6+/-0.44 microM) than the 25-amino acid linear CS-1 peptide. Cyclic peptides containing two more or one less methylene groups had similar potency to c(ILDV-NH(CH2)5CO) while a compound containing three less methylene groups, c(ILDV-NH(CH2)2CO), was inactive at 100 microM. 3. c(ILDV-NH(CH2)5CO) had little effect on cell adhesion mediated by two other integrins, VLA-5 (alpha5,beta1, CD49e/CD29) (K562 cell adhesion to fibronectin) or Leukocyte Function Associated molecule-1 (LFA-1, alphabeta2, CD11a/CD18) (U937 cell adhesion to Chinese hamster ovary cells transfected with intercellular adhesion molecule-1) at concentrations up to 300 microM. 4. c(ILDV-NH(CH2)5CO) inhibited ovalbumin delayed-type hypersensitivity or oxazolone contact hypersensitivity in Balb/c mice when dosed continuously from subcutaneous osmotic mini-pumps (0.1-10 mg kg(-1) day(-1)). Maximum inhibition (approximately 40%) was similar to that caused by the monoclonal antibody PS/2 (7.5 mg kg(-1) i.v.) directed against the alpha4 integrin subunit. 5. c(ILDV-NH(CH2)5CO) also inhibited oxazolone contact hypersensitivity when dosed intravenously 20 h after oxazolone challenge (1-10 mg kg(-1)). Ear swelling was reduced at 3 h and 4 h but not at 1 h and 2 h post-dose (10 mg kg(-1)). 6. Small molecule VLA-4 inhibitors derived from c(ILDV NH(CH2)5CO) may be useful as anti-inflammatory agents. PMID- 10372818 TI - Effects of A1-adenosine receptor antagonists on purinergic transmission in the guinea-pig vas deferens in vitro. AB - 1. Intracellularly recorded excitatory junction potentials (ej.ps) were used to study the effects of adenosine receptor antagonists on neurotransmitter release from postganglionic sympathetic nerve terminals in the guinea-pig vas deferens in vitro. 2. The A1 adenosine receptor antagonists, 8-phenyltheophylline (10 microM) and 8-cyclopentyl-1,3-dipropylxanthine (0.1 microM), increased the amplitude of e.j.ps evoked during trains of 20 stimuli at 1 Hz in the presence, but not in the absence, of the alpha2-adrenoceptor antagonist, yohimbine (1 microM) or the non selective alpha-adrenoceptor antagonist, phentolamine (1 microM). 3. Adenosine (100 microM) reduced the amplitude of e.j.ps, both in the presence and in the absence of phentolamine (1 microM). This inhibitory effect of adenosine is most likely caused by a reduction in transmitter release as there was no detectable change in spontaneous ej.p. amplitudes. 4. In the presence of phentolamine, application of the adenosine uptake inhibitor, S-(p-nitrobenzyl)-6-thioinosine (0.1 microM), had no effect on ej.p. amplitudes. 5. The phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine (100 microM), significantly increased the amplitudes of all e.j.ps evoked during trains of 20 stimuli at 1 Hz, both in the presence and in the absence of phentolamine (1 microM). 6. These results suggest that endogenous adenosine modulates neurotransmitter release by an action at prejunctional A1 adenosine receptors only when alpha2-adrenoceptors are blocked. PMID- 10372819 TI - The bradykinin B1 receptor and the central regulation of blood pressure in spontaneously hypertensive rats. AB - 1. We evaluated if the brain bradykinin (BK) B1 receptor is involved in the regulation of blood pressure (BP) in conscious rats. 2. Basal mean BP and HR were 115 +/- 2 and 165 +/- 3 mmHg and 345 +/- 10 and 410 +/- 14 beats min in Wistar Kyoto (WKY) and spontaneously hypertensive rats (SHR), respectively. Intracerebroventricular (i.c.v.) injection of 1 nmol B1 receptor agonist Lys desArg9-BK significantly increased the BP of WKY and SHR by 7+/-1 and 19+/-2 mmHg, respectively. One nmol Sar[D-Phe8]-desArg9-BK, a kininase-resistant B1 agonist, increased the BP of WKY and SHR by 19+/-2 and 17+/-2 mmHg, respectively and reduced HR in both strains. 3. I.c.v. injection of 0.01 nmol B1 antagonists, LysLeu8-desArg9-BK or AcLys[D-betaNal7,Ile8]-desArg9-BK (R715), significantly decreased mean BP in SHR (by 9+/-2 mmHg the former and 14+/-3 mmHg the latter compound), but not in WKY. In SHR, the BP response to R715 was associated to tachycardia. 4. I.c.v. Captopril, a kininase inhibitor, increased the BP of SHR, this response being partially prevented by i.c.v. R715 and reversed into a vasodepressor effect by R715 in combination with the B2 antagonist Icatibant. 5. I.c.v. antisense oligodeoxynucleotides (ODNs) targeted to the B1 receptor mRNA decreased BP in SHR, but not in WKY. HR was not altered in either strain. Distribution of fluorescein-conjugated ODNs was detected in brain areas surrounding cerebral ventricles. 6. Our results indicate that the brain B1 receptor participates in the regulation of BP. Activation of the B1 receptor by kinin metabolites could participate in the pathogenesis of hypertension in SHR. PMID- 10372820 TI - Diurnal variation in 5-HT1B autoreceptor function in the anterior hypothalamus in vivo: effect of chronic antidepressant drug treatment. AB - 1. Intracerebral microdialysis was used to examine the function of the terminal 5 hydroxytryptamine (5-HT) autoreceptor in the anterior hypothalamus of anaesthetized rats at two points in the light phase of the light-dark cycle. 2. Infusion of the 5-HT1A/1B agonist 5-methoxy-3-(1,2,3,6-tetrahydro-4-pyridyl)-1H indole (RU24969) 0.1, 1.0 and 10 microM through the microdialysis probe led to a concentration-dependent decrease (49, 56 and 65% respectively) in 5-HT output. The effect of RU24969 (1 and 5 microM) was prevented by concurrent infusion of methiothepin (1 and 10 microM) into the anterior hypothalamus via the microdialysis probe. Infusion of methiothepin alone (1.0 and 10 microM) increased (15 and 142% respectively) 5-HT output. 3. Infusion of RU24969 (5 microM) through the probe at mid-light and end-light resulted in a quantitatively greater decrease in 5-HT output at end-light compared with mid-light. 4. Following treatment with either paroxetine hydrochloride (10 mg kg(-1) i.p.) or desipramine hydrochloride (10 mg kg)(-1) i.p.) for 21 days the function of the terminal 5 HT1B autoreceptor was more markedly attenuated at end-light. 5. The data show that, as defined by the response to RU24969, the function of the 5-HT1B receptors that control 5-HT output in the anterior hypothalamus is attenuated following chronic desipramine or paroxetine treatment in a time-of-day-dependent manner. PMID- 10372821 TI - Role of Na+/Ca2+ exchange in endothelin-1-induced increases in Ca2+ transient and contractility in rabbit ventricular myocytes: pharmacological analysis with KB R7943. AB - 1. The effects of endothelin-1 (ET-1) on intracellular Ca2+ ion level and cell contraction were simultaneously investigated in rabbit ventricular cardiac myocytes loaded with indo-1/A1. The role of Na+/Ca2+ exchange in ET-1-induced positive inotropic effect (PIE) was examined by using KB-R7943 (2-[2-[4-(4 nitrobenzyloxy)phenyl]ethyl]isothiourea methanesulphonate), a selective inhibitor of reverse mode Na+/Ca2+ exchange. 2. ET-1 at 0.3 pM - 1 nM increased cell contraction and Ca2+ transient (CaT) with EC50 values of 2.9 pM and 1.2 pM, respectively, and the increase in amplitude of CaT was much smaller relative to the PIE: ET-1 at 1 nM increased peak cell shortening by 237%, while it increased peak CaT by 167%. For a given level of PIE, ET-1-induced increase in CaT was much smaller than that induced by elevation of [Ca2+]o and by isoprenaline. Therefore, ET-1 shifted the relationship between peak CaT and cell shortening to the left relative to the relationship for increase in [Ca2+]o, an indication that ET-1 increased myofibrillar Ca2+ sensitivity. 3. KB-R7943 at 0.1 microM and higher inhibited contraction and CaT induced by 0.1 nM ET-1 and at 0.3 microM it abolished the increase in CaT while inhibiting the PIE by 48.1%. Over concentration range of 0.1-0.3 microM, KB-R7943 neither inhibited baseline contraction and CaT nor the isoprenaline-induced response, although at 1 microM and higher it had a significant inhibitory action on these responses. 4. These results indicate that in rabbit ventricular myocytes both increases in CaT and myofibrillar Ca2+ sensitivity contribute to the ET-induced PIE, and the activation of reverse mode Na+/Ca2+ exchange may play a crucial role in increase in CaT induced by ET-1 in rabbit ventricular cardiac myocytes. PMID- 10372823 TI - Respective role of lipoxygenase and nitric oxide-synthase pathways in plasma histamine-induced macromolecular leakage in conscious hamsters. AB - 1. Intravital microscopy technique was used to determine the distribution of a fluorescent plasma marker (fluorescein-isothiocyanate-dextran, 150 kD; FD-150) into venular and interstitial compartments of dorsal skin fold preparations in conscious hamsters. 2. One mg kg(-1) histamine (i.v.) caused a biphasic decrease in venular fluorescence due to FD-150 extravasation in all organs (general extravasation). Immediately after injection, the venular fluorescence decreased and plateaued in 60 min. Ninety minutes after histamine injection, venular fluorescence further decreased until 180 min. Prior treatment with indomethacin (0.1 mg kg(-1), i.v.) did not modify the time-course of general extravasation but prevented histamine-induced venule dilatation. 3. Prior treatment with the 5 lipoxygenase activating protein (FLAP) inhibitor, 3-[1-(p-chlorobenzyl)-5 (isopropyl)-3-t-butylthioindol-2-yl]-2,2-d imethyl-propanoic acid sodium (MK 886)(10 microg kg(-1), i.v.), the leukotriene receptor antagonist, benzenemethanol a-pentyl-3-(2-quinolinylmethoxy) (REV-5901)(1 mg kg(-1), i.v.), or the glutathione-S-transferase inhibitor, ethacrynic acid (1 mg kg(-1), i.v.), delayed by 60 min the onset of general extravasation caused by 1 mg kg(-1) histamine. 4. Prior treatment with lipoxygenase pathway inhibitors and N(G)-nitro L-arginine-methylester (L-NAME)(100 mg kg(-1), i.v.) abolished the general extravasation and venule dilatation induced by 1 mg kg(-1) histamine. 5. Injection of 1 microg kg(-1) (i.v.), of leukotriene-C4 (LTC4) or -D4 (LTD4) induced immediate and sustained general extravasation and reduction in venule diameter, these effects being blocked by REV-5901. 6. Histamine (1 mg kg(-1), i.v.) induced biphasic decline in mean arterial blood pressure (MAP). An initial phase (from 0 to 60 min) was followed by a late phase beginning 90 min after histamine injection. L-NAME (100 mg kg(-1), i.v.) and aminoguanidine (1 mg kg( 1), i.v.) prevented the late phase of histamine-induced hypotension. 7. Thus, plasma histamine can trigger both an immediate cysteinyl-leukotriene (Cys-LT) dependent and a late nitric oxide (NO)-mediated inflammatory cascade. Although the cyclo-oxygenase (COX) pathway might account for histamine-induced venule dilatation, it would not influence histamine-induced extravasation. PMID- 10372822 TI - Adenosine mediates relaxation of human small resistance-like coronary arteries via A2B receptors. AB - 1. The receptor subtype and mechanisms underlying relaxation to adenosine were examined in human isolated small coronary arteries contracted with the thromboxane A2 mimetic, 1,5,5-hydroxy-11alpha, 9alpha-(epoxymethano)prosta-5Z, 13E-dienoic acid (U46619) to approximately 50% of their maximum contraction to K+ (125 mM) depolarization (Fmax). Relaxations were normalized as percentages of the 50% Fmax contraction. 2. Adenosine caused concentration-dependent relaxations (pEC50, 5.95+/-0.20; maximum relaxation (Rmax), 96.7+/-1.4%) that were unaffected by either combined treatment with the nitric oxide inhibitors, NG-nitro-L arginine (L-NOARG; 100 microM) and oxyhaemoglobin (HbO; 20 microM) or the ATP dependent K+ channel (KATP) inhibitor, glibenclamide (10 microM). The pEC50 but not Rmax to adenosine was significantly reduced by high extracellular K+ (30 mM). Relaxations to the adenylate cyclase activator, forskolin, however, were unaffected by high K+ (30 mM). 3. Adenosine and a range of adenosine analogues, adenosine, 2-chloroadenosine (2-CADO), 5'-N-ethyl-carboxamidoadenosine (NECA), R( )-N6-(2-phenylisopropyl)-adenosine (R-PIA), S(+)-N6-(2-phenylisopropyl)-adenosine (S-PIA), N6-cyclopentyladenosine (CPA), 1-deoxy-1-[6-[[(3 iodophenyl)methyl]amino]-9H-purin-9-yl]-N-methyl-beta- D-ribofuranuronamide (IB MECA), 2-p-(2-carboxyethyl)phenethylamino-5'-N-ethylcarboxamido adenosine hydrochloride (CGS 21680), relaxed arteries with a rank order of potency of NECA= 2-CADO >adenosine= IB-MECA = R-PIA= CPA > S-PIA)> CGS 21680. 4. Sensitivity but not Rmax to adenosine was significantly reduced approximately 80 and 20 fold by the non-selective adenosine receptor antagonist, 8-(p-sulphophenyl)theophylline (8-SPT) and the A2 receptor antagonist, 3,7-dimethyl-1-propargylxanthine (DMPX). By contrast, the A1-selective antagonist, 1,3-dipropyl-8-cyclopentylxanthine (DPCPX) had no effect on pEC50 or Rmax to adenosine. 5. These results suggest that A2B receptors mediate relaxation to adenosine in human small coronary arteries which is independent of NO but dependent in part on a K+-sensitive mechanism. PMID- 10372824 TI - Effects of prolonged cold storage on double peaked vasoconstrictor responses to periarterial nerve stimulation in isolated canine splenic arteries. AB - 1. P2X-Purinoceptors and alpha1-adrenoceptors have previously been shown to involve in the double peaked vasoconstrictor responses to periarterial electrical nerve stimulation in the isolated and perfused canine splenic artery. The present study made an attempt to investigate effects of prolonged cold storage (7 days at 4 degrees C) on vasoconstrictor responses to periarterial electrical nerve stimulation, tyramine, noradrenaline and adenosine 5'-triphosphate (ATP) in the isolated canine splenic artery. 2. The periarterial nerve stimulation (1-10 Hz) readily causes a double peaked vasoconstriction in the non-stored preparations. After cold stored for 7 days, the double peaked vasoconstriction was still recognized, although the response became significantly smaller. The first phase was decreased relatively greater than the second phase by the cold storage. 3. In the cold stored preparations, the dose-response curve for tyramine was shifted to the right in a parallel manner. Prazosin almost completely inhibited tyramine induced vasoconstriction but alpha,beta-methylene ATP failed to influence the response to tyramine. 4. The vasoconstrictor responses to noradrenaline and ATP were not significantly modified by the prolonged cold storage. 5. From these results, it is concluded that the functions of sympathetic co-transmission of purinergic components might be influenced more than that of adrenergic components in the cold storage canine splenic artery. PMID- 10372825 TI - Rapid actions of calcitriol and its side chain analogues CB1093 and GS1500 on intracellular calcium levels in skeletal muscle cells: a comparative study. AB - 1. The ability of synthetic analogues of the secosteroid hormone 1alpha,25 dihydroxy-vitamin-D3 [calcitriol, CT; 1,25(OH)2D3] to exert non-genomic (rapid) effects on target cells has been scarcely studied. To evaluate the pharmacological potential of the CT side-chain analogues CB1093 and GS1500, we compared their fast effects on intracellular calcium concentration ([Ca2+]i) in chick skeletal muscle cells with those elicited by the natural hormone. 2. Both analogues, similarly to CT, specifically induced rapid (30-60 s) and sustained rises in [Ca2+]i levels. CB1093 and GS1500 were more potent than the natural hormone at concentrations as low as 10(-13) M (4.5 fold stimulation) and 10(-12) M (2.5 fold), respectively, whereas higher concentrations (10(-9)- 10(-8) M) of CT were more effective than the analogues in elevating [Ca2+]i. Cyclic AMP was markedly increased by both analogues pointing for a role of this messenger in the fast actions of the synthetic compounds. 3. In Ca2+ free medium CT and analogues elicited a transient elevation in [Ca2+]i. The PLC inhibitors U73122 (2 microM) and neomycin (0.5 mM), as well as depletion of intracellular stores with thapsigargin (1 microM), completely prevented CB1093/GS1500-dependent changes in [Ca2+]i suggesting that, similarly to CT, these analogues mobilized Ca2+ from an IP3/thapsigargin-sensitive store. 4. The voltage-dependent calcium channel (VDCC) blocker nifedipine (2 microM) reduced by 50-60% the influx phase of the [Ca2+]i response to CB1093 and GS1500, indicating that VDCC contributed partially to Ca2+ entry. The Ca2+ readdition protocol suggested that analogue-dependent activation of a SOC entry pathway accounted, to the same extent as for CT, for the remaining non-VDCC mediated Ca2+ influx. PMID- 10372826 TI - Ex vivo assay to determine the cyclooxygenase selectivity of non-steroidal anti inflammatory drugs. AB - 1. In this study we describe experiments to establish ex vivo the selectivity of non-steroidal anti-inflammatory drugs (NSAIDs) given in vivo. 2. Anaesthetised (Inactin, 120 mg kg(-1)) male Wistar rats (220-250 g) received an i.v. dose of one of the following compounds (dose mg kg(-1)): aspirin (20), diclofenac (3), L 745,337 (30), nimesulide (15), salicylate (20), sulindac (10). Blood samples were taken before and up to 6 h after dosing and the plasma obtained from it was tested for its ability to inhibit prostanoid formation in IL-1beta-treated A549 cells (COX-2 system) and human washed platelets (COX-1 system). For control the same compounds were also added directly to the assay systems. 3. All drugs, except sodium salicylate, inhibited COX-1 and COX-2 when added directly to the test systems. Plasma from aspirin-treated rats was without effect on either COX-1 or COX-2, consistent with the rapid in vivo metabolism to salicylate. Conversely, plasma from sulindac-treated rats inhibited COX-1 and COX-2 with potencies according with in vivo metabolism to sulindac sulphide. Diclofenac was COX-1/2 non-selective when tested in vitro, but a slightly preferential inhibitor of COX 2 when tested ex vivo. Nimesulide was confirmed as preferential inhibitor of COX 2 both in vitro and ex vivo. L-745,337 was a selective COX-2 inhibitor when tested in vitro or ex vivo. 4. In conclusion, our experiments show clearly (a) NSAIDs inactivation, (b) activation of prodrugs, and (c) NSAIDs selectivity. Our assay provides useful information about the selectivity of NSAIDs that could be extended by the analysis of plasma samples taken from humans similarly treated with test drugs. PMID- 10372827 TI - Effects of the endogeneous cannabinoid, anandamide, on neuronal activity in rat hippocampal slices. AB - 1. The arachidonic acid derivative arachidonylethanolamide (anandamide) is an endogeneous ligand of cannabinoid receptors that induces pharmacological actions similar to those of cannabinoids such as delta9-tetrahydrocannabinol (THC). We examined whether anandamide can influence excessive neuronal activity by investigating stimulation-induced population spikes and epileptiform activity in rat hippocampal slices. For this purpose, the effects of anandamide were compared with those of the synthetic cannabinoid agonist WIN 55,212-2 and its inactive S( )-enantiomer WIN 55,212-3. 2. Both anandamide (1 and 10 microM) and WIN 55,212-2 (0.1 and 1 microM) decreased the amplitude of the postsynaptic population spike and the slope of the field excitatory postsynaptic potential (field e.p.s.p.) without affecting the presynaptic fibre spike of the afferents. At a concentration of 1 microM, WIN 55,212-2 completely suppressed the postsynaptic spike, whereas the S(-)-enantiomer WIN 55,212-3 produced only a slight depression. The CB1 receptor antagonist SR 141716 blocked the inhibition evoked by the cannabinoids. SR 141716 had a slight facilitatory effect on neuronal excitability by itself. 3. Anandamide shifted the input-output curve of the postsynaptic spike and the field e.p.s.p. to the right and increased the magnitude of paired-pulse facilitation indicating a presynaptic mechanism of action. 4. Anandamide and WIN 55,212-2, but not WIN 55,212-3, attenuated both stimulus-triggered epileptiform activity in CA1 elicited by omission of Mg2+ and spontaneously occurring epileptiform activity in CA3 elicited by omission of Mg2+ and elevation of K+ to 8 mM. The antiepileptiform effect of these cannabinoids was blocked by SR 141716. 5. In conclusion, cannabinoid receptors of the CB1 type as well as their endogeneous ligand, anandamide, are involved in the control of neuronal excitability, thus reducing excitatory neurotransmission at a presynaptic site, a mechanism which might be involved in the prevention of excessive excitability leading to epileptiform activity. PMID- 10372828 TI - Effects of intrathecal administration of nitric oxide synthase inhibitors on carrageenan-induced thermal hyperalgesia. AB - 1. We examined the effects of various nitric oxide synthase (NOS) inhibitors on carrageenan-induced thermal hyperalgesia. 2. First, we determined the time point at which a subcutaneous plantar injection of carrageenan into the rat hindpaw produced maximum thermal hyperalgesia. Subsequently, we demonstrated that intrathecal administration of the non-selective NOS inhibitor L-N(G)-nitro arginine methyl ester (L-NAME) produces a dose-dependent reduction of carrageenan induced thermal hyperalgesia. 3. Four relatively selective NOS inhibitors were then tested for their efficacy at reducing carrageenan-induced thermal hyperalgesia. Initially, the effects of prolonged treatment with inhibitors of neuronal [7-nitroindazole (7-NI) and 3-bromo-7-nitroindazole (3-Br)] and inducible [aminoguanidine (AG) and 2-amino-5,6-dihydro-methylthiazine (AMT)] NOS were examined. All agents were injected three times intrathecally during the course of inflammation caused by the plantar injection of carrageenan, and thermal hyperalgesia was measured at 6 h post-carrageenan using a plantar apparatus. 4. All inhibitors, except for 7-NI, were effective at attenuating the carrageenan-induced thermal hyperalgesia when compared with vehicle treatment. 5. Finally, the effects of early versus late administration of neuronal and inducible NOS inhibitors on carrageenan-induced thermal hyperalgesia were examined. We found that neither 3-Br nor AG significantly affected thermal hyperalgesia when administered during the early phase of carrageenan inflammation, while only AG was able to reduce thermal hyperalgesia when administered during the late phase of the injury. 6. Our results suggest that inducible NOS contributes to thermal hyperalgesia in only the late stages of the carrageenan-induced inflammatory response, while neuronal NOS likely plays a role throughout the entire time course of the injury. PMID- 10372829 TI - Direct inhibition of the N-methyl-D-aspartate receptor channel by dopamine and (+)-SKF38393. AB - 1. Dopamine is known to modulate glutamatergic synaptic transmission in the retina and in several brain regions by activating specific G-protein-coupled receptors. We have examined the possibility of a different type of mechanism for this modulation, one involving direct interaction of dopamine with ionotropic glutamate receptors. 2. Ionic currents induced by fast application of N-methyl-D aspartate (NMDA) were recorded under whole-cell patch-clamp in cultured striatal, thalamic and hippocampal neurons of the rat and in retinal neurons of the chick. Dopamine at concentrations above 100 microM inhibited the NMDA response in all four neuron types, exhibiting an IC50 of 1.2 mM in hippocampal neurons. The time course of this inhibition was fast, developing in less than 100 ms. 3. The D1 receptor agonist (+)-SKF38393 mimicked the effect of dopamine, with an IC50 of 58.9 microM on the NMDA response, while the enantiomer (-)-SKF38393 was ineffective at 50 microM. However, the D1 antagonist R(+)-SCH23390 did not prevent the inhibitory effect of (+)-SKF38393. 4. The degree of inhibition by dopamine and (+)-SKF38393 depended on transmembrane voltage, increasing 2.7 times with a hyperpolarization of about 80 mV. The voltage-dependent block by dopamine was also observed in the presence of MgCl2 1 mM. 5. Single-channel recordings showed that the open times of NMDA-gated channels were shortened by (+)-SKF38393. 6. These data suggested that the site to which the drugs bound to produce the inhibitory effect was distinct from the classical D1-type dopamine receptor sites, possibly being located inside the NMDA channel pore. It is concluded that dopamine and (+)-SKF38393 are NMDA channel ligands. PMID- 10372830 TI - Enhancement of vascular permeability by specific activation of protease-activated receptor-1 in rat hindpaw: a protective role of endogenous and exogenous nitric oxide. AB - 1. To clarify the role of the first thrombin receptor/protease-activated receptor (PAR)-1 in an inflammatory process, we tested and characterized the effect of intraplantar (i.pl.) administration of the highly specific PAR-1 agonist TFLLR NH2 in rat hindpaw. 2. TFLLR-NH2 administered i.pl. at 0.01-0.03 micromol per paw enhanced vascular permeability in the hindpaw and produced paw oedema in a dose dependent manner. This effect was almost completely abolished by repeated pretreatment with compound 48/80 to deplete inflammatory mediators in mast cells. 3. The NO synthase inhibitor N(G)-nitro-L-arginine methyl ester or N-iminoethyl-L ornithine, preadministered i.pl., stereospecifically potentiated the i.pl. TFLLR NH2-induced permeability increase, while the NO donor sodium nitroprusside or NOC 18, given i.pl., suppressed the effect of TFLLR-NH2. 4. These findings demonstrate that specific activation of PAR-1 produces increased vascular permeability accompanied by oedema formation in the rat hindpaw, predominantly via mast cell degranulation, and that endogenous and exogenous NO plays a protective role in the PAR-1-mediated inflammatory event. PMID- 10372831 TI - A comparison of the inhibitory activity of PDE4 inhibitors on leukocyte PDE4 activity in vitro and eosinophil trafficking in vivo. AB - 1. Phosphodiesterase (PDE) 4 inhibitors have been shown to inhibit eosinophil PDE4 activity in vitro and accumulation of eosinophils in experimental airways inflammation. However, direct effects on eosinophil trafficking have not been studied in detail and it is not known if activity in vitro translates into efficacy in vivo. In the present study, we compared the activity of five PDE4 inhibitors in vitro and against trafficking of (111)In-eosinophils in cutaneous inflammation in the guinea-pig. 2. The rank order of potency for inhibition of PDE4 activity in guinea-pig eosinophil, neutrophil and macrophage, and human neutrophil lysates was RP73401 > SB207499 >CDP840 > rolipram > LAS31025. On TNFalpha production by human PBMC, all inhibitors with the exception of rolipram showed potency similar to their effect on neutrophil lysates. 3. In a brain cerebellum binding assay, the rank order of potency at displacing [3H]-rolipram was RP73401 > rolipram > SB207499 > CDP840 > LAS30125. 4. Trafficking of (111)In eosinophils to skin sites injected with PAF, ZAP or antigen in sensitized sites was inhibited by oral administration of all PDE4 inhibitors. The rank order of potency was RP73401 = rolipram > LAS31025 > SB207499 > CDP840. 5. With the exception was RP73401, which was the most potent compound in all assays, there was no clear relationship between activity of PDE4 inhibitors in vitro and capacity to inhibit eosinophil trafficking in vivo. Thus, we conclude that in vitro activity of PDE4 inhibitors does not predict in vivo efficacy in an experimental model of eosinophil trafficking. PMID- 10372833 TI - Introduction to renal osteodystrophy: calcium metabolism in health and uremia. PMID- 10372832 TI - Activation of nitric oxide synthase by beta 2-adrenoceptors in human umbilical vein endothelium in vitro. AB - 1. Some animal studies suggest that beta-adrenoceptor-mediated vasorelaxation is in part mediated through nitric oxide (NO) release. Furthermore, in humans, we have recently shown that forearm blood flow is increased by infusion of beta2 adrenergic agonists into the brachial artery, and the nitric oxide synthase (NOS) inhibitor N(G)-monomethyl-L-arginine (L-NMMA) inhibits this response. 2. The purpose of the present study was to determine whether stimulation of human umbilical vein endothelial beta-adrenoceptors causes vasorelaxation and nitric oxide generation, and whether this might be mediated by cyclic adenosine-3',5' monophosphate (cyclic AMP). 3. Vasorelaxant responses were determined in umbilical vein rings to the nonselective beta-adrenergic agonist isoprenaline and to the cyclic AMP analogue dibutyryl cyclic AMP, following precontraction with prostaglandin F2alpha. 4. NOS activity was measured in cultured human umbilical vein endothelial cells (HUVEC) by the conversion of [3H]-L-arginine to [3H]-L citrulline, and adenylyl cyclase activity by the conversion of [alpha-32P]-ATP to [32P]-cyclic AMP. 5. Isoprenaline relaxed umbilical vein rings, and this vasorelaxation was abolished by beta2- (but not beta1-) adrenergic blockage, and by endothelium removal or 1 mM L-NMMA. In addition, vasorelaxant responses to dibutyryl cyclic AMP were inhibited by 1 mM L-NMMA, with a reduction in Emax from 90.0+/-9.3% to 50.5+/-9.9% (P<0.05). 6. Isoprenaline 1 microM increased NOS activity in HUVEC (34.0+/-5.9% above basal, P<0.001). Furthermore, isoprenaline increased adenylyl cyclase activity in a concentration-dependent manner; this response was inhibited by beta2 (but not beta1-) adrenergic blockade. Forskolin 1 microM and dibutyryl cyclic AMP 1 mM each increased NOS activity in HUVEC, to a degree similar to isoprenaline 1 microM. The increase in L-arginine to L citrulline conversion observed with each agent was abolished by coincubation with NOS inhibitors. 7. These results indicate that endothelial beta2-adrenergic stimulation and cyclic AMP elevation activate the L-arginine/NO system, and give rise to vasorelaxation, in human umbilical vein. PMID- 10372834 TI - Modulation of parathyroid cell function by calcium ion in health and uremia. AB - Extracellular calcium ion concentration is the major determinant of parathyroid hormone (PTH) secretion from parathyroid cells. In dialysis patients with secondary hyperparathyroidism, higher calcium concentration is needed to suppress PTH secretion as demonstrated by the PTH-calcium curve. Such abnormal sensitivity to extracellular calcium ion has been recently explained by the decrease in number of calcium-sensing receptors, especially on cells in nodular hyperplasia, which is the advanced type of parathyroid hyperplasia in uremia. Modulation of the sensitivity of parathyroid cells to calcium has become partly possible through the use of newly developed calcimimetics. PMID- 10372835 TI - Vitamin D and the parathyroid. AB - Vitamin D's biologically active metabolite, 1,25(OH)2D3, has important effects upon the parathyroid cell that are relevant to both the physiology of mineral metabolism and the regulation of the secondary hyperparathyroidism of chronic renal failure. 1,25(OH)2D3 markedly decreases parathyroid hormone (PTH) gene transcription and thus PTH synthesis and secretion. It also acts to decrease parathyroid cell proliferation. Nonhypercalemic analogs of 1,25(OH)2D3 are being developed that may have a wider therapeutic window than 1,25(OH)2D3 itself. In the situations of chronic hypocalcemia and hypophosphatemia, there are interesting interrelationships between 1,25(OH)2D3 and the post-transcriptional regulation of the PTH gene. In nodular secondary hyperparathyroidism, there is down-regulation of the vitamin D receptor in the parathyroid. Different vitamin D receptor genotypes may be associated with higher levels of serum PTH and a predisposition to autonomous hyperplasia. PMID- 10372836 TI - The role of phosphorus in the development of secondary hyperparathyroidism and parathyroid cell proliferation in chronic renal failure. AB - Hyperplasia of the parathyroid glands and high levels of parathyroid hormone (PTH) are among the most consistent findings in patients with chronic renal failure. In early renal failure, alterations in vitamin D metabolism play a key role in the development of secondary hyperparathyroidism. Low levels of calcitriol and decreased expression of the vitamin D responsive element may allow greater synthesis and secretion of PTH. Phosphorus independent of serum calcium and calcitriol increases PTH synthesis and secretion by a post-transcriptional mechanism. Studies in vivo in uremic rats demonstrated that an increase in dietary phosphorus induces parathyroid gland hyperplasia. If the rats are then fed a low-phosphorus diet, the levels of serum PTH return to normal; however, the size of the parathyroid glands remains enlarged. No apoptosis was observed in the glands. To further characterize the effects of phosphorus on PTH synthesis and secretion, intact rat parathyroid glands were metabolically labeled during a 4 hour incubation in methionine-free medium containing 1.25 mM Ca2+, [35S]methionine, and either 2.8 mM or 0.2 mM phosphorus. Total PTH secretion, as measured in the medium, was increased more than 6-fold in glands incubated in high-phosphorus medium compared with glands incubated in the low-phosphorus medium. Thus, in the past 20 years, numerous investigators have provided strong evidence for the action of phosphorus on PTH secretion. Unfortunately, the absence of a parathyroid cell line is slowing the progress in understanding the molecular mechanism(s) involved in phosphorus regulation of PTH. PMID- 10372837 TI - Cell biology of parathyroid hyperplasia in uremia. AB - Marked parathyroid hyperplasia of heterogeneous degrees is often seen in chronic dialysis patients with severe secondary hyperparathyroidism. In uremia, parathyroid cell proliferation is initially stimulated by decreased concentration of calcium ions and calcitriol and also by direct effect of phosphate accumulation, leading to diffuse hyperplasia of the parathyroid. Then, small nodules caused by monoclonal cell proliferation form within diffuse hyperplasia, which progress to form nodular hyperplasia. Cells in nodular hyperplasia have a lower density of calcitriol receptor and calcium-sensing receptor than diffuse hyperplasia and are thus more resistant to medical therapy, including calcitriol pulse therapy. One of these nodules may grow more vigorously than the others and may finally occupy a large part of the enlarged gland. Genetic mutations and rearrangements of these cells in nodular hyperplasia remain to be fully elucidated in the near future to establish an effective method for the prevention of parathyroid hyperplasia in uremia. PMID- 10372838 TI - Medical management of secondary hyperparathyroidism in uremia. AB - The prevention and treatment of the secondary hyperparathyroidism of chronic renal failure by medical means relies on a number of different possible approaches, which should be tailored to each patient's individual needs. Schematically, prevention should start early during the course of chronic renal failure (ie, when plasma intact parathyroid hormone is normal or only slightly elevated). Small calcium supplements prevent the development of a calcium deficit and may prevent parathyroid hormone oversecretion. The various therapeutic options that are available at present include: oral or intravenous vitamin D and vitamin D derivatives, in particular the 1alpha-hydroxylated vitamin D compounds; oral calcium supplements (calcium carbonate and calcium acetate) to avoid calcium depletion and also to bind phosphate in the intestinal lumen; aluminum-containing phosphate binders, the use of which should be restricted; oral magnesium salts (magnesium carbonate and magnesium hydroxide), which often are not well tolerated; general measures, such as dietary restriction of phosphate intake; and, in the case of resistance to all these approaches, the possibility of attempting ultrasound-guided ethanol injection of grossly hyperplastic parathyroid glands. Finally, it is encouraging to know that new drugs are in development, including calcium-free, aluminum-free, nonabsorbable oral phosphate binders, potentially nonhypercalcemic vitamin D derivatives, and calcimimetics. Some of them already have entered the stage of clinical evaluation, and preliminary results are promising. PMID- 10372839 TI - Surgical management of secondary hyperparathyroidism in uremia. AB - Advanced secondary (renal) hyperparathyroidism (HPT) induced by uremia is one of the most serious complications for long-term hemodialysis patients. Parathyroidectomy (PTx) is indicated in patients with severely advanced renal HPT that is refractory to medical treatment, including calcitriol pulse therapy. The clinical effect of PTx is striking. However, skeletal deformity, vessel calcification, and remarkable reduction of bone content is irreversible. Therefore, it is important to perform PTx at the right time. Based on histopathological and pathophysiological investigations, nodular hyperplasia is monoclonal neoplasia with abnormal parathyroid hormone response to extracellular calcium and vitamin D. When parathyroid hyperplasia progresses to nodular hyperplasia, PTx should be required. Total PTx with forearm autograft is the preferred procedure for renal HPT, especially for patients who need to continue hemodialysis treatment after PTx. Removal of all parathyroid glands, including supernumerary glands, at the initial operation and proper choice of adequate parathyroid tissue for autograft are important to prevent persistent and recurrent HPT. In this series of 782 patients, the function of autografted parathyroid tissue is almost satisfactory and no retransplantation of cryopreserved parathyroid tissue was necessary; however, graft-dependent recurrent HPT was not negligible. In conclusion, total PTx with forearm autograft is very effective and adequate treatment for advanced renal HPT and parathyroid function can be controlled after PTx. PMID- 10372840 TI - Prevention of renal osteodystrophy in predialysis patients. AB - Impaired calcitriol synthesis is one of the major factors contributing to the development of secondary hyperparathyroidism in patients with chronic renal failure. Vitamin D therapy, particularly 1alpha-hydroxyvitamin D3, even in low doses, has been shown to be effective in the treatment of secondary hyperparathyroidism in patients with mild-to-moderate chronic renal failure. Complications associated with calcitriol and alfacalcidol therapy, which include hypercalcemia and progressive deterioration of renal function, have been reported in some patients. The majority of the studies reviewed, however, demonstrated that daily calcitriol and alfacalcidol doses below 0.25 microg are rarely associated with hypercalcemia, hyperphosphatemia, or progressive decline in renal function. In addition, these complications usually resolve with the reduction in dose or discontinuation of the medication. Thus, vitamin D therapy may be valuable in the treatment of patients with mild-to-moderate chronic renal failure who may be at high risk of developing secondary hyperparathyroidism. PMID- 10372841 TI - Relative hypoparathyroidism and adynamic bone disease. AB - Renal bone disease results in significant morbidity in patients with end-stage renal failure. Renal osteodystrophy is a mixture of different conditions with different pathogenetic factors involved. Most recently a new form of renal bone disease, adynamic bone disease, has emerged as the most frequent finding on bone biopsy of patients on dialysis therapy. The etiology of this new entity is not fully understood, but relatively low levels of intact serum parathyroid hormone are frequently associated with this disorder and may play an important role in its pathogenesis. PMID- 10372842 TI - Pathogenesis and management of dialysis-related amyloid bone disease. AB - Dialysis-related amyloidosis (DRA) is a major complication of chronic renal failure and long-term renal replacement therapy. Beta2-Microglobulin is a major constituent of amyloid fibrils in DRA. Amyloid deposition can present as carpal tunnel syndrome, destructive arthropathy, or subchondral bone erosions and cysts. A definitive diagnosis of DRA can only be made using histological findings, but various analytical imaging methods often support diagnosis. Therapy of an established DRA is limited to symptomatic approaches and surgical removal of amyloid deposits. High-flux biocompatible dialysis membranes can be used to delay DRA development. Recent studies have suggested a pathogenic role for a new modification of beta2-microglobulin in DRA. Increased carbonyl compounds modify proteins, which leads to the augmentation of advanced glycation and lipoxidation end products. Thus, uremia might be a state of carbonyl overload with potentially damaging proteins, leading to a new modification of beta2-microglobulin in amyloid fibrils and development of DRA. PMID- 10372843 TI - "Crack" cocaine-induced syndrome mimicking sarcoidosis. AB - A 39-year-old man with a history of frequent "crack" cocaine use of several years' duration presented with progressive dyspnea. Evaluation revealed bilateral interstitial pulmonary infiltrates and hilar adenopathy, diffuse pulmonary uptake of gallium, and markedly elevated serum angiotensin-converting enzyme activity. Open lung biopsy revealed interstitial and perivascular collections of histiocytes containing refractile, polarizable material, presumably inhaled along with the cocaine. Paratracheal lymph nodes were enlarged, reactive, and contained similar polarizable material. The well-formed, non-necrotizing granulomata characteristic of sarcoidosis were not present in either tissue specimen. To our knowledge, the association of chronic crack cocaine inhalation with this constellation of clinical findings, typically seen in sarcoidosis, has not previously been described. PMID- 10372844 TI - Hepatitis A-induced diabetes mellitus, acute renal failure, and liver failure. AB - A 38-year-old otherwise healthy man presented with hepatic failure (aspartate aminotransferase of 7212 U/L, alanine aminotransferase of 6629 U/L, total and direct bilirubin of 10.7 mg/dL) and acute renal failure (creatinine of 11.6 mg/dL and blood urea nitrogen of 42 mg/dL), which required hemodialysis when the creatinine increased to 21 mg/dL, with a blood urea nitrogen of 115 mg/dL, and the patient became oliguric. On admission, this patient also had a lipase of 1833 U/L, amylase of 211 U/L, glucose of 210 mg/dL, and reactive IgM antibody for acute hepatitis A. The hepatitis and acute renal failure resolved in 3 months, but this patient continues to have type II diabetes mellitus 7 years after the hepatitis A infection. This case illustrates that hepatitis A infection may be severe with liver failure, acute renal failure, and permanent diabetes mellitus as sequale of this infection. PMID- 10372845 TI - Primary antiphospholipid syndrome and cerebral atrophy: a rare association? AB - Neurologic complications are common in patients with antiphospholipid syndrome. In this article, we report the case of a young woman with neurologic disorders, a history of hypertension and transient ischemic attacks, and cerebral atrophy associated with primary antiphospholipid syndrome (PAPS). Magnetic resonance imaging of the brain showed multiple ischemic lesions and remarkable atrophy of frontal and parietal lobes. Cerebral atrophy in patients with PAPS can be considered as a feature of this disease. The case is discussed on the basis of relevant past literature. Although there are few reports on neuroradiologic findings in patients with PAPS, cerebral atrophy has been described. Because PAPS is more frequently recognized today than in the past, this condition should be included in the differential diagnosis of cerebral atrophy, particularly in young patients. PMID- 10372846 TI - Assessment and importance of personality disorders in medical patients: an update. AB - BACKGROUND: Personality disorders in medical patients have received less attention than depression, anxiety, or somatization. METHOD: We conducted a selective literature search to assess the role of personality disorders in medical patients. RESULTS: Review of recent studies suggests a high prevalence and morbidity of personality disorders in medical populations. Important correlates in selected groups are depression, somatization, noncompliance, sexual risk taking, and substance abuse. Difficulties in physician-patient relationships are also frequently reported. Psychiatric interventions are considered beneficial, though no single treatment of choice is available. CONCLUSIONS: We recommend that physicians consider the possibility of personality disorders in medical patients to choose appropriate treatments for selected symptoms. Training in interviewing skills may enhance recognition of personality disorders and management of associated psychiatric conditions. PMID- 10372847 TI - Mesenteric venous thrombosis. AB - BACKGROUND: Mesenteric venous thrombosis is an uncommon but often lethal form of intestinal ischemia. METHODS: We reviewed pertinent literature on mesenteric venous thrombosis using MEDLINE search. RESULTS: We found that previous abdominal surgery and hypercoagulable states are the most common conditions associated with mesenteric venous thrombosis. The symptoms and signs related to mesenteric venous thrombosis are not specific. In the majority of cases, the diagnosis is established by a high index of clinical suspicion and noninvasive imaging techniques. Immediate operation is indicated if signs of peritonitis or intestinal infarction are present. Administration of heparin is beneficial for reducing recurrence and mortality. CONCLUSION: Clinicians should consider the possibility of acute mesenteric venous thrombosis when faced with a patient having abdominal pain out of proportion to the physical findings and with a negative workup for the common causes of abdominal pain. PMID- 10372848 TI - Patellar tendon ruptures. AB - BACKGROUND: Isolated rupture of the patellar tendon is a rare injury. Often occurring during a fall in 20- and 30-year olds, patients may have a preexisting medical condition (eg, history of steroid use) or a history of repetitive microtrauma to the knee. A high-riding patella on physical examination and radiographs is pathognomonic. METHODS: Immediate orthopaedic referral for surgical repair is necessary to reestablish knee extension. Delay in diagnosis can make surgical treatment more difficult. Current methods of postoperative rehabilitation are evolving. RESULTS: Evaluative studies based on rating scales show satisfactory clinical and functional results after surgery. However, time lost from work and recreation may be protracted, and quadriceps atrophy is often evident. CONCLUSIONS: Ruptures of the patellar tendon should be diagnosed acutely and immediately referred to an orthopaedic surgeon. The impact of the injury to the patient may be long-standing even after operative treatment. Contemporary surgical and rehabilitative techniques give the best opportunity for restoration of functional activity. PMID- 10372849 TI - Centennial year of Ronald Ross' epic discovery of malaria transmission: an essay and tribute. AB - Malaria has been in existence since ancient times and for millennia thought to be carried by vapors. Alphonse Laveran made the discovery of the malarial parasite in 1880. The next most important milestone in the history of malaria was the unraveling of the mystery of malaria transmission. Ronald Ross, born in British India and educated as a doctor in England, was working in the Indian Medical Service when he discovered the transmission of malarial parasites by mosquitoes in 1897. The next year, he completed the life cycle of malarial parasites in birds. In celebration of the centennial year of that epic discovery, we reminisce on that event, focusing on the discoverer. PMID- 10372850 TI - Potential association between calcified thoracic lymphadenopathy due to previous Histoplasma capsulatum infection and pulmonary Mycobacterium avium complex disease. AB - BACKGROUND: Among patients with pulmonary disease due to Mycobacterium avium complex (MAC) seen recently at our center, a substantial number have had extensive calcified mediastinal, hilar, and peribronchial lymphadenopathy, a finding historically inconsistent with pulmonary MAC disease. METHOD: We retrospectively studied the frequency of calcified lymphadenopathy in the chest and prevalence of known risk factors for MAC infection in 79 patients with pulmonary MAC disease who were referred to our hospital over a 1-year period. RESULTS: Calcified intrathoracic adenopathy was present in 25 of the 79 patients (32%). Residential histories revealed that 20 of the 25 patients (80%) with such calcified chest adenopathy reported living for substantial periods in the regions indigenous for Histoplasma capsulatum. In contrast, the residences of patients without calcified chest adenopathy were more evenly distributed throughout the country. Nineteen of these 25 patients (76%) with calcified chest adenopathy had no known predisposing risk factor for the infection; in contrast, the proportion of patients with no calcified adenopathy who also had no identifiable classic risk factor tended to be lower (32/54, 59%). CONCLUSION: In this retrospective study, we observed that (1) a large number of patients with pulmonary MAC disease had no identifiable risk factor, (2) calcified chest adenopathy was present in one third of the patients, (3) the residential history of those with calcified adenopathy mirrored the endemic region of histoplasmosis, and, (4) conversely, those patients with pulmonary MAC who lived outside the histoplasmosis belt had no such adenopathy. Thus, we hypothesize that previous fungal infection may predispose the lungs of certain patients to subsequent invasion by MAC, presumably by airway distortion and/or parenchymal damage. PMID- 10372851 TI - Motivating the obese child to move: the role of structured exercise in pediatric weight management. AB - BACKGROUND: The prevalence of childhood obesity is rapidly increasing. Successful prevention and treatment of childhood obesity depends on increasing the physical activity patterns of obese youth. However, motivating the obese child to participate in physical activity is difficult. METHODS: We designed a four-phase physical activity intervention, consisting of a structured progressive exercise program of moderate intensity, along with motivational methods to increase physical activity and improve body movement awareness. RESULTS: Seventy-three overweight children participated in the weight management program. They had a significant weight loss and reduction in body fat, which was maintained at 1-year follow-up. Subjects also maintained lean body mass and showed improved physical activity patterns. CONCLUSIONS: Progressive exercise, used in conjunction with nutrition and behavior modification, provides successful motivational strategies. These strategies encourage increased physical activity patterns, the adoption of regular structured exercise training, and the loss of excess body fat. PMID- 10372852 TI - Inclusion of resistance exercise in a multidisciplinary outpatient treatment program for preadolescent obese children. AB - BACKGROUND: Safe and effective exercise programs are needed to prevent and treat chronic diseases in childhood. In particular, preadolescent obese children should participate in activities that are specific to their special needs. METHODS: We included a moderate intensity, progressive resistance training program in a multidisciplinary weight management program for obese preadolescent children. The program included diet, behavior modification, and aerobic and flexibility exercises. RESULTS: The subjects reported no accidents or injuries and 100% compliance with the minimum required exercise prescription. Weight, percent ideal body weight, body mass index, and percent fat were reduced significantly at 10 weeks and did not increase significantly at 1 year follow-up. Height and lean body mass increased significantly at 1 year. CONCLUSION: A resistance training program may be safely included in a multidisciplinary weight management program for obese preadolescent children. PMID- 10372853 TI - Diabetes-related lower-extremity amputations disproportionately affect Blacks and Mexican Americans. AB - BACKGROUND: We sought to identify the age-adjusted incidence of lower-extremity amputation (LEA) in Mexican Americans, blacks, and non-Hispanic whites with diabetes in south Texas. METHODS: We summarized medical records for hospitalizations for LEAs for 1993 in six metropolitan statistical areas in south Texas. RESULTS: Age-adjusted incidence per 10,000 patients with diabetes was 146.59 in blacks, 60.68 in non-Hispanic whites, and 94.08 in Mexican Americans. Of the patients, 47% of amputees had a history of amputation, and 17.7% were hospitalized more than once during 1993. Mexican Americans had more diabetes related amputations (85.9%) than blacks (74.7%) or non-Hispanic whites (56.3%). CONCLUSIONS: This study is the first to identify the incidence of diabetes related lower-extremity amputations in minorities using primary data. Minorities had both a higher incidence and proportion of diabetes-related, LEAs compared with non-Hispanic whites. Public health initiatives and national strategies, such as Healthy People 2000 and 2010, need to specifically focus on high-risk populations and high-risk geographic areas to decrease the frequency of amputation and reamputation. PMID- 10372854 TI - Effect of injected versus iontophoretic corticosteroid on the rabbit tendon. AB - BACKGROUND: The etiologic role of corticosteroid therapy in tendon rupture is controversial. This study compared the effects of injected versus iontophoretically delivered corticosteroid on the normal rabbit Achilles tendon. METHODS: Rabbits were divided into three treatment groups: (1) corticosteroid injections, (2) iontophoretically delivered corticosteroid, and (3) no treatment. One tendon of each rabbit in the treatment groups was treated with either drug injection or iontophoresis; the tendon of the other leg served as a control. Some tendons were used for testing elastic modulus, ultimate load, and ultimate stress, while the remaining tendons were evaluated histologically. RESULTS: Injections of either corticosteroid or saline into the tendon sheath resulted in short-term changes in tendon biomechanical characteristics and somewhat higher histologic severity scores; however, iontophoretic delivery of corticosteroid or saline did not affect either significantly. CONCLUSIONS: Iontophoresis using sterile water or corticosteroid resulted in minimal or no biochemical and histologic changes in the tendon compared with injection of either substance. The method of corticosteroid delivery may be as important as the actual drug effects on the biomechanical and histologic properties of tendons. PMID- 10372855 TI - Delusions about the Internet. AB - As computer use increases throughout the world, more people are becoming familiar with computers, the Internet, and the roles that both play in their daily lives. There have already been reports of people becoming addicted to the Internet, and we now report on two cases of men who had delusions that they were controlled by and entwined with the Internet. We feel that this phenomenon is not a new diagnostic entity but more likely a new subtype of previously reported psychiatric illnesses. We also discuss the genesis of delusional content and how topics covered in the popular media may influence delusional content. PMID- 10372856 TI - Squamous cell carcinoma arising in the epithelial lining of a dentigerous cyst. AB - The dentigerous cyst is a common oral lesion arising as a developmental anomaly during amelogenesis. In rare instances, the epithelial lining of these cysts may give rise to squamous cell carcinoma (SCC). Fewer than 50 cases of this rare entity have been reported in the world literature to date. We present an additional case of SCC arising in a dentigerous cyst with a rationale for our treatment approach. In addition, we offer a review of the literature and a review of the clinical, histologic, and radiographic findings associated with this finding. We discuss epidemiologic data and treatment options. PMID- 10372858 TI - Abuse history: is it really important in the medical encounter? AB - The negative effects of a history of severe abuse as a child are becoming increasingly well known to the physician and, in the adult survivor, include both mental health and physical sequelae. Intervention for emotional and psychologic consequences can include a mental health referral. Awareness of physical sequelae can aid the physician in diagnosis and intervention. However, the impact of severe childhood abuse in the medical encounter does not end with diagnosing and managing somatic complaints and/or providing a mental health referral. Severe childhood abuse directly affects the physician-patient relationship by causing the predictable difficulties that adult survivors of severe abuse experience in any of their intimate relationships. PMID- 10372857 TI - Waxing and waning gynecomastia: an indication of noncompliant use of prescribed medication. AB - We present two cases of recurrent gynecomastia in men enrolled in a placebo controlled trial evaluating the efficacy of finasteride in treating benign prostatic hyperplasia. When the pharmacologic records were examined, it was apparent that the breast tissue hyperplasia diminished when the patients become noncompliant with their study medication and then resumed therapy. Because of the difficulty in obtaining accurate data on an individual's ability to maintain a consistent pharmacologic regimen, we believe that observing such "waxing and waning gynecomastia" may provide the physician with a clue regarding a patient's actual compliance with certain medications. PMID- 10372859 TI - Fluoroquinolone-induced tendinopathy: what do we know? AB - Fluoroquinolones are relatively safe, effective antibiotics. As their use becomes more frequent, so will the adverse side effects. I highlight a rare but debilitating adverse reaction-fluoroquinolone-induced tendinopathy. Case reports and letters from 1987 to 1998 were identified by using Grateful Med and PubMed Internet accesses to the National Library of Medicine. Articles were reviewed for clinical practicality. There are few articles on fluoroquinolone-induced tendinopathy in the US literature targeting primary care physicians. This entity has been described in many case reports, but little has been done to isolate the causative agents. Incidence of this side effect is difficult to estimate, since no prospective studies are available for review or calculation of risk. Fluoroquinolone-induced tendinopathy appears more commonly in tendons under high stress. The cause is probably multifactorial. Risk factors for the development of fluoroquinolone-induced tendinopathy are age, renal failure, corticosteroid use, and previous tendinopathy from fluoroquinolones. PMID- 10372860 TI - Portal hypertension due to extensive hepatic cysts in autosomal dominant polycystic kidney disease. AB - Liver cysts are a well-recognized feature of autosomal dominant polycystic kidney disease (ADPKD) and occur in 77% of patients more than 60 years old. Serious sequelae, however, are rare, the two most common complications being pain and cyst infections. Portal hypertension has been reported in ADPKD due to the rare presence of congenital hepatic fibrosis. We report a case of ADPKD in a patient who had portal hypertension due to distortion of portal vein and venules by extensive hepatic cysts. PMID- 10372861 TI - Renal cell carcinoma in an intrathoracic kidney: radiographic findings and surgical considerations. AB - Ectopic intrathoracic kidney is a rare phenomenon and is usually an incidental finding on a chest radiograph. Of all intrathoracic kidneys, congenital ectopia is most often shown, with a traumatic etiology occurring much less frequently. We report a case of an ectopic intrathoracic kidney with associated renal cell carcinoma. Management, which was based on current treatment recommendations for isolated renal masses, consisted of radical nephrectomy. The patient has been without evidence of disease recurrence for 36 months after surgery. PMID- 10372862 TI - Spiritual resonance: pursuit in the bluegrass state. PMID- 10372863 TI - Graduate medical education: a "handful" of tasks. PMID- 10372864 TI - Review of the update of personality disorders. PMID- 10372865 TI - The future we want; the future we get. PMID- 10372866 TI - Kinematic and qualitative analysis of lower-extremity movements in preterm infants with brain lesions. AB - BACKGROUND AND PURPOSE: The purposes of this study were to evaluate the effects of preterm birth, severe brain lesions, and postterm age on kicking movements of young infants and to compare the prognostic value of kinematic analysis of kicking with a qualitative assessment of infants' spontaneous movements. SUBJECTS: The subjects were 12 full-term infants without brain injury, 12 low risk preterm infants without brain injury, and 11 preterm infants with severe brain lesions (periventricular leukomalacia). METHODS: Videotape recordings of each infant's motor behavior in a supine position were made at 1 and 3 months postterm age. Kicking frequency, temporal organization of the kick cycle, coordination among different joints, and interlimb coordination were measured. A qualitative assessment for lower-extremity movements and a Gestalt judgment of general movement quality according to Prechtl's method were made from the same videotape recordings. RESULTS: Kinematic analysis showed only mild differences among the 3 groups of infants. Qualitative assessment of the lower-extremity movements, however, showed that preterm infants with brain lesions, and particularly those who later were found to have cerebral palsy, consistently had fewer segmental movements of the foot and abnormal general movements at both ages. CONCLUSION AND DISCUSSION: The data suggest that the mechanisms responsible for kicking movements in newborns and young infants do not appear to be influenced by the extrauterine environment or by brain lesions, at least at the ages studied. Qualitative assessment of lower-extremity and general movements seems to be more appropriate for clinical purposes. PMID- 10372867 TI - Accuracy of digitization using automated and manual methods. AB - BACKGROUND AND PURPOSE: Computerized 3-dimensional (3-D) motion measurement systems are used by those interested in human motion. The purposes of this study were (1) to determine the limits of accuracy in determining intersegmental angles during pendular motion at varying speeds and (2) to determine changes in accuracy introduced by autodigitization and digitization by experienced manual raters. METHODS: Angular speed of a T-shaped pendulum was systematically increased by releasing the pendulum from 4 angles (0 degrees [no movement], 45 degrees, 90 degrees, and 120 degrees). Twelve reference angles calculated from markers placed on the pendulum were estimated over 20 frames for 10 trials at each release position. RESULTS: Mean errors across trials and frames for intersegmental angles reconstructed by a 3-D motion measurement system were within +/- 1 degree across all release positions. An analysis of variance and a post hoc Tukey test revealed that the mean error for the autodigitized trials was larger than that for the manually digitized trials. For the autodigitized trials, the static trials (release position=0 degrees) produced less mean error than the trials with movement produced. The ICCs showed a high degree of consistency among all raters, ranging from .707 to .999. CONCLUSION AND DISCUSSION: Our findings support the conclusion that under carefully controlled conditions, a 3-D motion measurement system can produce clinically acceptable measurements of accuracy across a range of angular speeds. Furthermore, acceptable accuracy is possible regardless of the digitization method. PMID- 10372868 TI - The clinical doctorate: a framework for analysis in physical therapist education. AB - This article explores major considerations for analysis and discussion of the role of the clinical doctorate as the first professional degree in physical therapist education (DPT). A process for this analysis is posed based on a conceptual framework developed by Stark, Lowther, Hagerty, and Orczyk through grounded theory research on professional education. External influences from society and the profession, institutional and programmatic influences, and articulation of critical dimensions of professional competence and professional attitudes as major categories are discussed in relation to the DPT. A series of questions generated from the application of the model are put forth for continued discussion and deliberation concerning the DPT. We conclude that the DPT provides the best pathway to serve society, the patient, and the profession. PMID- 10372869 TI - Diagnosis of intermittent vascular claudication in a patient with a diagnosis of sciatica. AB - BACKGROUND AND PURPOSE: The purpose of this case report is to illustrate the importance of medical screening to rule out medical problems that may mimic musculoskeletal symptoms. CASE DESCRIPTION: This case report describes a woman who was referred with a diagnosis of sciatica but who had signs and symptoms consistent with vascular stenosis. The patient complained of bilateral lower extremity weakness with her pain intensity at a minimal level in the region of the left sacroiliac joint and left buttock. She also reported numbness in her left leg after walking, sensations of cold and then heat during walking, and cramps in her right calf muscle. She did not report any leg pain. A medical screening questionnaire revealed an extensive family history of heart disease. Examination of the lumbar spine and nervous system was negative. A diminished dorsalis pedis pulse was noted on the left side. Stationary cycling in lumbar flexion reproduced the patient's complaints of lower-extremity weakness and temporarily abolished her dorsalis pedis pulse on the left side. OUTCOMES: She was referred back to her physician with a request to rule out vascular disease. The patient was subsequently diagnosed, by a vascular specialist, with a "high grade circumferential stenosis of the distal-most aorta at its bifurcation." DISCUSSION: This case report points out the importance of a thorough history, a medical screening questionnaire, and a comprehensive examination during the evaluation process to rule out medical problems that might mimic musculoskeletal symptoms. PMID- 10372870 TI - Hypermobility syndrome. PMID- 10372871 TI - Guide to physical therapist practice: revisions. American Physical Therapy Association. PMID- 10372872 TI - Small incision external levator repair: technique and early results. AB - PURPOSE: To describe a new surgical technique and early results of external levator repair performed through a small skin incision. METHODS: A chart review of consecutive patients undergoing small incision external levator repair was conducted. This modified external levator repair was performed through an 8-mm eyelid crease incision. Patients with unilateral or bilateral aponeurogenic blepharoptosis were candidates for the technique. Patients with excessive horizontal upper eyelid laxity and those requiring blepharoplasty in addition to blepharoptosis surgery did not undergo this technique. Patients who underwent previous upper eyelid surgery or concurrent brow surgery were excluded from the review. Preoperative measurements included upper eyelid margin reflex distance, levator function, and degree of dermatochalasis, as well as Goldmann visual field results. Outcome measures included incidence and type of intraoperative complications, postoperative upper eyelid position (including margin reflex distance, eyelid contour, and symmetry), incidence and type of postoperative complications, and revisions or additional necessary surgery. RESULTS: Twenty eight eyelids of 17 patients met study inclusion criteria. Preoperative margin reflex distance +/- SD averaged 0.8 +/- 0.4 mm. Average length of follow-up was 28 +/- 5 weeks (range, 15 to 52 weeks). No significant intraoperative complications occurred. Postoperative margin reflex distance averaged 3.7 +/- 0.3 mm. Two eyelids were mildly undercorrected, and one demonstrated moderately peaked contour postoperatively. Satisfactory eyelid position and contour were achieved in 25 of 28 treated eyelids. No patient elected reoperation. CONCLUSIONS: Early results demonstrated that small incision levator repair is safe and generally effective. This minimally invasive external levator repair is useful for a carefully selected subset of patients with aponeurogenic blepharoptosis. PMID- 10372873 TI - Effects of peribulbar anesthesia on ocular blood flow in patients undergoing cataract surgery. AB - PURPOSE: The effects of extraconal, peribulbar anesthesia on ocular blood flow may be caused by concomitant elevations in intraocular pressure or direct pharmacologic alteration of vascular tone. We quantified the effect on ocular circulation with a new technique for assessment of ocular hemodynamics. METHODS: In a prospective study, ocular hemodynamics were measured before and 1 and 5 minutes after peribulbar anesthesia in 22 eyes with age-related cataract. Measurements included fundus pulsation amplitude with a laser interferometric method assessing the pulsatile choroidal blood flow and mean blood flow velocity as well as resistive index in the ophthalmic and central retinal artery with Doppler sonography. Systemic blood pressure and pulse were monitored throughout the period of ocular hemodynamic measurements. RESULTS: Fundus pulsation amplitude decreased significantly after peribulbar anesthesia (after 1 minute and 5 minutes: -13% and -8%; P < .001). In the central retinal artery, mean blood flow velocity dropped (-15%; P < .001) and resistive index increased (+3%; P = .02) 1 minute after peribulbar anesthesia compared with baseline. There were no changes in ophthalmic artery hemodynamics. Intraocular pressure was elevated 1 minute after peribulbar anesthesia (+29%; P = .003) but reached baseline values after 5 minutes. CONCLUSION: Pulsatile choroidal blood flow and retinal blood flow velocities were reduced after peribulbar anesthesia. These reductions were still present 5 minutes after peribulbar anesthesia, when intraocular pressure had returned to baseline values. This supports the theory of drug-induced vasoconstriction after peribulbar anesthesia. A loss of vision may be a risk of peribulbar anesthesia in patients who have compromised ocular blood flow before surgery. PMID- 10372874 TI - Population-based assessment of the outcome of cataract surgery in an urban population in southern India. AB - PURPOSE: To assess the outcome of cataract surgery in an urban population in southern India. METHODS: As part of a population-based cross-sectional epidemiologic study, the Andhra Pradesh Eye Disease Study, 2,522 people of all ages, including 1,399 individuals 30 years of age or older, from 24 clusters representative of the population of Hyderabad in southern India underwent a detailed interview and ocular evaluation including logarithm of minimal angle of resolution (logMAR) visual acuity, refraction, slit-lamp biomicroscopy, applanation tonometry, gonioscopy, dilation, cataract grading, aphakia/pseudophakia status, and stereoscopic fundus evaluation. Automated threshold visual fields and slit-lamp and fundus photography were performed when indicated by standardized criteria. Very poor outcome in an eye that had undergone cataract surgery was defined as presenting distance visual acuity worse than 20/200, and poor outcome was defined as visual acuity worse than 20/60 to 20/200. RESULTS: In subjects 50 years of age or older, after adjustment for age and sex distribution, the rate of having had cataract surgery in one or both eyes was 14.6% (95% confidence interval [CI], 11.4% to 17.8%). Of 131 eyes (91 subjects) that had undergone cataract surgery, 28 (21.4%; 95% CI, 14.4% to 28.4%) had very poor outcome and another 40 (30.5%; 95% CI, 22.6% to 38.4%) had poor outcome. The very poor outcome in 20 (71.4%) of 28 eyes and poor outcome in 23 (57.5%) of 40 eyes could be attributed to surgery-related causes or inadequate refractive correction. With multivariate analysis, very poor outcome as a result of surgery-related causes or inadequate refractive correction was more likely to be associated with intracapsular cataract extraction than with extracapsular cataract extraction (odds ratio, 9.34; 95% CI, 2.49 to 35.06) in subjects belonging to the lowest socioeconomic status (odds ratio, 4.92; 95% CI, 1.16 to 20.93) and with date of surgery 3 or fewer years before the survey than with more than 3 years (odds ratio, 4.52; 95% CI, 1.33 to 15.39). Also, very poor or poor outcome as a result of surgery-related causes or inadequate refractive correction was associated with women (odds ratio, 2.55; 95% CI, 1.06 to 6.16). CONCLUSIONS: The very high rate of very poor and poor visual outcome, predominantly as a result of surgery-related causes and inadequate refractive correction, in this urban population of India suggests that more attention is needed to improve the visual outcome of cataract surgery. In order to deal with cataract-related visual impairment in India, as much emphasis on surgical quality, refractive correction, and follow-up care is necessary as on the number of surgeries. PMID- 10372876 TI - Three-year clinical outcome after penetrating keratoplasty for keratoconus with the guided trephine system. AB - PURPOSE: To determine the long-term clinical outcome after keratoplasty with the guided trephine system in keratoconus eyes. METHODS: In a prospective study, all consecutive cases of penetrating keratoplasty had trephination performed with the guided trephine system, with which both donor and recipient cornea are trephined from the epithelial side with a same-sized blade. For wound closure, a double running antitorque suture technique with 10-0 nylon was used. Uncorrected and best-corrected Snellen visual acuity, subjective refraction, and astigmatism by keratometry were evaluated after final suture removal, 2 and 3 years postoperatively. RESULTS: In the 31 patients (31 eyes) enrolled, mean best corrected visual acuity improved from 0.72 +/- 0.16 (20/30) after final suture removal to 0.88 +/- 0.15 (20/25) 3 years postoperatively (P < .001). The mean spherical equivalent increased from -0.86 +/- 2.10 diopters after final suture removal to -2.35 +/- 2.65 diopters 3 years postoperatively (P < .001). Mean keratometric astigmatism decreased from 4.68 +/- 1.76 diopters after final suture removal to 3.57 +/- 1.37 diopters 3 years postoperatively (P = .001). Furthermore, an increase in mean keratometric levels with time (P = .01) was observed and associated with myopic shift (r(s) = -.46, P = .008). CONCLUSION: With the guided trephine system, we attained favorable visual results, with prolonged improvement of visual acuity during the entire follow-up period. Our data show low and decreasing degrees of corneal astigmatism over time. During the follow-up period, a myopic shift was found after final suture removal. Nevertheless, this technique of performing same-sized grafts reduces postoperative residual myopia. PMID- 10372875 TI - Tear mixing under a soft contact lens: effects of lens diameter. AB - PURPOSE: Tear exchange under a soft contact lens is modest, and higher exchange rates may be necessary to reduce extended-wear complications; what is not known is the optimal soft lens design to increase tear mixing. We explored the effect of lens diameter on tear mixing. METHODS: Twenty-three subjects wore four different soft contact lenses with diameters of 12.0, 12.5, 13.0, and 13.5 mm. Tear mixing was quantified by placing fluorescein isothiocyanate-dextran on the posterior lens surface, inserting the lens, and monitoring the changes in fluorescence intensity in the postlens tear film. Tear mixing, expressed as the percentage decrease in fluorescence intensity per blink, was estimated using an exponential model. Lens movement was videotaped and lens comfort was graded on a 50-point scale (50 = excellent comfort). Subjects reporting a comfort level of less than 35 were excluded. RESULTS: The mean +/- SE tear mixing rates were 1.82% +/- 0.17%, 1.61% +/- 0.16%, 1.34% +/- 0.17%, and 1.24% +/- 0.17% per blink for the 12.0-, 12.5-, 13.0-, and 13.5-mm diameter lenses, respectively. By regression analysis we found that, on average, mixing under the 12.0-mm lens was 0.59% per blink greater than with the 13.5-mm lens (P = .0024). Lens diameter was a significant predictor of lens comfort, and adjusting for the effects of comfort weakened the relationship between diameter and tear replenishment rate, although the mean rate under the 12.0-mm lens was still 0.43% per blink greater than with the 13.5-mm lens (P = .0468). CONCLUSIONS: These data suggest that smaller diameter soft lenses provide substantially better tear mixing than larger lenses; however, even small lenses provide modest tear mixing compared with rigid contact lenses. PMID- 10372877 TI - Relationship between parapapillary atrophy and visual field abnormality in primary open-angle glaucoma. AB - PURPOSE: To investigate the relationship of parapapillary atrophy measured by confocal scanning laser ophthalmoscopy to visual field sensitivity measured with standard automated perimetry and short-wavelength automated perimetry in patients with primary open-angle glaucoma. METHODS: Forty-seven eyes of 47 primary open angle glaucoma patients with increased intraocular pressure (> or = 22 mm Hg) were enrolled. Optic nerve head topography and parapapillary atrophy (beta and alpha zones) were assessed by confocal scanning laser ophthalmoscopy. Mean deviation and corrected pattern SD were assessed with standard automated perimetry and short-wavelength automated perimetry. RESULTS: Beta and alpha zones were found in 23 (49%) and 47 (100%) eyes with primary open-angle glaucoma, respectively. The area of beta zone showed significant correlations with MD of standard automated perimetry, corrected pattern SD of standard automated perimetry, and corrected pattern SD of short-wavelength automated perimetry (Spearman r = -0.366, P = .012; r = 0.327, P = .025; and r = 0.436, P = .002, respectively). The area of alpha zone showed a significant correlation with mean deviation of standard automated perimetry (r = -0.378, P = .009). Mean MD of standard automated perimetry, mean corrected pattern SD of standard automated perimetry, and mean corrected pattern SD of short-wavelength automated perimetry were significantly worse in eyes with beta zone than in eyes without beta zone. CONCLUSIONS: Parapapillary atrophy measured by confocal scanning laser ophthalmoscopy, especially beta zone, is associated with glaucomatous visual field loss demonstrated by standard automated perimetry and short-wavelength automated perimetry. PMID- 10372878 TI - Fluorescein angiographic abnormalities after prophylactic macular photocoagulation for high-risk age-related maculopathy. AB - PURPOSE: Initial studies suggest that drusen associated with age-related maculopathy resolve in response to laser photocoagulation; there are conflicting reports regarding whether this treatment might prevent neovascular complications and blindness. The goal of the Drusen Laser Study is to maintain good visual acuity in eyes at the highest risk for neovascular complications of age-related maculopathy. In this report, we alert the ophthalmic community to possible laser induced complications in patients treated within the context of this clinical trial. METHODS: A double-masked, randomized, controlled clinical trial of prophylactic macular photocoagulation for high-risk age-related maculopathy is in progress. Patients randomly assigned to treatment received a ring-type distribution of 12 light spots of argon laser photocoagulation. Drusen were treated directly only if they were present at the protocol treatment locations. Fluorescein angiography was performed in all patients at yearly review, and at nonprotocol visits if symptoms or clinical examination were suggestive of choroidal neovascularization. RESULTS: Fluorescein angiographic abnormalities suggestive of choroidal neovascularization have been seen in treated eyes only: one patient in the pilot study and six patients in the Drusen Laser Study. No fluorescein angiographic abnormalities were seen in eyes of control subjects. CONCLUSIONS: Laser photocoagulation in high-risk age-related maculopathy may induce choroidal neovascularization and, therefore, is not recommended outside the context of a randomized, controlled clinical trial. PMID- 10372879 TI - Patterns of diabetic macular edema with optical coherence tomography. AB - PURPOSE: We report cross-sectional images of diabetic macular edema and correlation between tomographic features and visual acuity with best correction by means of optical coherence tomography. METHOD: In a prospective study, optical coherence tomography was performed in 59 eyes of 42 patients with diabetic macular edema and in 10 eyes of 10 normal control subjects. RESULTS: Optical coherence tomography showed three patterns of structural changes in diabetic macular edema: sponge-like retinal swelling (52 [88%] of 59 eyes), cystoid macular edema (28 [47%] of 59 eyes), and serous retinal detachment (9 [15%] of 59 eyes). Some eyes had more than one pathologic change. Retinal swelling was more pronounced in the outer rather than the inner retinal layers. Cystoid macular edema was located mainly in the outer retinal layers. In eyes with long-standing cystoid macular edema, cystoid spaces had fused, resulting in a large cystoid cavity involving almost the entire retinal layer. Hard exudates were seen as highly reflective areas located in the outer retinal layers. The retinal thickness at the central fovea and the visual acuity with best correction showed an intermediate negative correlation in eyes without cystoid macular edema (correlation coefficient: -0.61, P < .01). CONCLUSIONS: Diabetic macular edema involved three structural changes, including sponge-like retinal swelling (88%), cystoid macular edema (47%), and serous retinal detachment (15%). Visual acuity with best correction moderately correlated with retinal thickness regardless of the different tomographic features. PMID- 10372880 TI - Antioxidant enzymes in the macular retinal pigment epithelium of eyes with neovascular age-related macular degeneration. AB - PURPOSE: To test the hypothesis that neovascular age-related macular degeneration is related to oxidative stress involving the macular retinal pigment epithelium. This study investigated, as a function of age, levels of enzymes that defend tissues against oxidative stress in the macular retinal pigment epithelium of human eyes with this disease. METHODS: Surgical specimens of macular choroidal neovascular membranes from eyes with age-related macular degeneration and the macular regions of whole donor eyes with neovascular age-related macular degeneration or without evident ocular disease were studied by quantitative electron microscopic immunocytochemistry with colloidal gold-labeled second antibodies. Relative levels in retinal pigment epithelium cell cytoplasm and lysosomes were determined of five enzymes believed to protect cells from oxidative stress, as well as levels of the retinal pigment epithelium marker cytoplasmic retinaldehyde-binding protein, for comparison with the enzymes. RESULTS: Copper, zinc superoxide dismutase immunoreactivity increased and catalase immunoreactivity decreased with age in cytoplasm and lysosomes from macular retinal pigment epithelium cells of normal eyes and eyes with age-related macular degeneration. Cytoplasmic retinaldehyde-binding protein immunoreactivity showed no significant relationship to age or the presence of neovascular age related macular degeneration. Glutathione peroxidase immunoreactivity was absent from human retinal pigment epithelium cells. Both heme oxygenase-1 and heme oxygenase-2 had highly significantly greater immunoreactivity in retinal pigment epithelium cell lysosomes than in cytoplasm, differing from the much greater cytoplasmic immunoreactivity of the other proteins studied. This immunoreactivity decreased with age, particularly in the lysosomes of retinal pigment epithelium cells from eyes with neovascular age-related macular degeneration. These decreases were of borderline significance (P = .067 for heme oxygenase-1; P = .12 for heme oxygenase-2) when eyes with age-related macular degeneration were compared with normal eyes by multivariable logistic regression. CONCLUSIONS: The high heme oxygenase-1 and heme oxygenase-2 lysosomal antigen levels in macular retinal pigment epithelium cells of eyes with neovascular age-related macular degeneration suggest that oxidative stress causes a pathologic upregulation of these enzymes. Increased lysosomal disposal may indicate that the reparative functions of these enzymes are accompanied by deleterious effects, necessitating their rapid removal from the cell. The much higher heme oxygenase-1 and heme oxygenase-2 antigen levels in macular retinal pigment epithelium cells from younger individuals suggest that protective mechanisms against oxidation and, hence, presumably to the development of age-related macular degeneration, decrease with age. PMID- 10372881 TI - Malignant transformation of an optic disk melanocytoma. AB - PURPOSE: To report a case of malignant transformation of an optic disk melanocytoma with a second melanocytoma in the ciliary body. METHODS: Clinical data including visual acuity, visual fields, color fundus photographs, fluorescein angiogram, and ultrasonogram and histopathologic studies of this case were reviewed. RESULTS: The right eye of a 65-year-old white woman was diagnosed with melanocytoma of the optic nerve. Four years later, the tumor became significantly larger. The best-corrected visual acuity declined from 20/40 to counting fingers and the size of the tumor increased fourfold in 2 years. The right globe was enucleated. Histopathologic studies demonstrated moderately pigmented spindle-B malignant melanoma cells adjacent to and within a population of large, polyhedral, heavily pigmented melanocytoma cells that extended to the lamina cribrosa and optic nerve. There was also a deeply pigmented melanocytoma in the ciliary body. CONCLUSION: This is a rare case of malignant melanoma transformed from an optic disk melanocytoma. Periodic follow-up of the patient with optic disk melanocytoma is necessary. PMID- 10372882 TI - The dilemma of ophthalmic changes spreads to Asia. PMID- 10372884 TI - Acanthamoeba keratitis with live isolates treated with cryosurgery and fluconazole. AB - PURPOSE: To report live, active trophozoites in an eye with Acanthamoeba keratitis that resembled Acanthamoeba polyphagia in the anterior chamber fluid obtained by transcorneal tap. METHOD: After prediagnostic therapy had failed, we performed cryosurgery to break the corneal cell walls and treated the patient with oral fluconazole. RESULTS: The condition resolved after 8 weeks of oral fluconazole therapy. Residual leukoma was treated by corneal graft. CONCLUSION: Live, motile Acanthamoeba can be isolated from an anterior chamber tap; combination therapy with oral fluconazole after corneal cryosurgery may be effective. PMID- 10372883 TI - Fluorescein test for the detection of striae in the corneal flap after laser in situ keratomileusis. AB - PURPOSE: To report a technique for detecting striae in the corneal flap after laser in situ keratomileusis. METHODS: Fluorescein dye was instilled in the eye, and the patient was asked to blink. The tear film was examined at the slit lamp with the cobalt filter 1 or 2 seconds after blinking. RESULTS: The uneven pattern of pooling in the tear film was a sensitive indicator of the presence of striae in the flap. CONCLUSION: This technique may be useful in detecting minimal striae in the corneal flap in patients with unexplained suboptimal visual acuity after laser in situ keratomileusis. PMID- 10372885 TI - Herpes simplex virus in the trabeculum of an eye with corneal endotheliitis. AB - PURPOSE: To report an eye with corneal endotheliitis and increased intraocular pressure in which the trabeculum demonstrated immunoreactivity for herpes simplex virus. METHOD: Case report. A 62-year-old man presented with increased intraocular pressure, keratic precipitates, and corneal stromal edema in his left eye. The tissue excised during trabeculectomy was immunohistochemically examined for herpetic viruses. RESULT: Immunoreactivity for herpes simplex virus was identified in the trabeculum. CONCLUSION: Herpes simplex virus may cause trabeculitis and increased intraocular pressure in patients with corneal endotheliitis. PMID- 10372886 TI - Effects of contact lenses on scanning laser polarimetry of the peripapillary retinal nerve fiber layer. AB - PURPOSE: To determine the effects of contact lenses on scanning laser polarimetry of the peripapillary nerve fiber layer. METHODS: In a prospective study using the Nerve Fiber Analyzer (Laser Diagnostic Technologies, San Diego, California), retinal nerve fiber layer thickness in 22 subjects (51 eyes) was imaged with and without contact lenses (disposable and nondisposable daily wear soft and rigid gas permeable). Measurements of the circumference and of each quadrant were compared using paired Student t test. RESULTS: Nerve Fiber Analyzer measurements with and without contact lenses were not significantly different for any of the contact lens types tested (P > or = .11), using either hyperopic (to +4 diopters) or myopic (to -8.5 diopters) lenses. CONCLUSION: Contact lens wear and refractive power of the eye within the range tested do not significantly affect scanning laser polarimetry of the peripapillary nerve fiber layer. PMID- 10372887 TI - Effects of methotrexate treatment on serum immunoreactivity of a patient with normal-pressure glaucoma. AB - PURPOSE: Increased serum immunoreactivity to retinal proteins may have a role in the disease process of some glaucoma patients. We describe a patient with normal pressure glaucoma whose serum immunoreactivity to retinal proteins regressed after methotrexate treatment for rheumatoid disease. METHOD: Case report. RESULTS: In a 66-year-old white female with normal-pressure glaucoma and rheumatoid disease, sequential Western blots using patient sera against retinal proteins demonstrated a decrease in the immunoreactive bands after treatment. During the treatment period of 3 years, her visual fields appeared to have improved. Optic disk examination during the short periods without treatment, however, disclosed new, bilateral splinter hemorrhages on the optic disks. CONCLUSION: These observations suggest a potential role for immune-based intervention in similar patients. PMID- 10372888 TI - Multiple evanescent white dot syndrome in older patients. AB - PURPOSE: To report two patients in their seventh decade who exhibited findings consistent with multiple evanescent white dot syndrome. METHODS: Case reports of two patients referred for evaluation of decreased vision, visual field loss, and retinal white spots. RESULTS: A 60-year-old man and a 67-year-old woman had photopsia, visual field loss, and decreased central visual acuity. Examination disclosed numerous white retinal spots, ranging from 50 to 400 microm, with eventual foveal granularity. Visual field testing showed an enlarged blind spot and peripheral field defects. Fluorescein angiography, electroretinography, and electrooculography results were consistent with multiple evanescent white dot syndrome. Eventually, the retinal lesions resolved in both patients and baseline visual acuity was recovered. CONCLUSION: A diagnosis of multiple evanescent white dot syndrome should be considered in patients with retinal findings typical of multiple evanescent white dot syndrome, regardless of age. PMID- 10372889 TI - Laser pointer maculopathy. AB - PURPOSE: To report a case of macular damage from a laser pointer. METHOD: Case report. A 19-year-old woman had an acute reduction of visual acuity in the right eye after deliberately staring into a commercial class 2 laser pointer for approximately 10 seconds. RESULTS: The patient's best-corrected visual acuity was RE: 20/40, and she had two small pericentral scotomata, as well as a hypopigmented ring-shaped lesion in the fovea. Within 8 weeks, her visual acuity improved to 20/20 and visual field returned to normal, but a subjective relative decrease in brightness of objects viewed by the right eye was apparent. Retinal pigment epithelial abnormality persisted. CONCLUSIONS: Commercial laser pointers, commonly used for teaching and entertainment purposes, may cause notable macular damage if abused. Morphologically, this may manifest as foveal retinal pigment epithelial disturbance. PMID- 10372890 TI - Spontaneous separation of an idiopathic macular pucker in a young girl. AB - PURPOSE: To report a young girl with spontaneous separation of an idiopathic epiretinal membrane and notable visual recovery. METHOD: Case report. RESULT: A 12-year-old girl had spontaneous improvement in visual acuity of the left eye from 20/100 to 20/40 attributable to spontaneous peeling of an idiopathic epiretinal membrane more than 2 years after it was diagnosed. CONCLUSION: Conservative treatment can be considered in young patients with epiretinal membrane because spontaneous separation may occur and result in good vision. PMID- 10372891 TI - Modified sutureless sclerotomies in pars plana vitrectomy. AB - PURPOSE: To study the effectiveness and safety of a modified sutureless sclerotomy technique in pars plana vitrectomy. METHODS: We rotated the scleral tunnels of the original sutureless sclerotomy technique through 90 degrees, thus rendering them parallel to the corneoscleral limbus. This modified technique was applied to 25 consecutive eyes (25 patients) that had pars plana vitrectomy during a 2-month period. RESULTS: Twenty (80%) of 25 eyes (25 patients) did not require suturing of the sclerotomy sites associated with pars plana vitrectomy. Eight (11%) of 75 sclerotomy sites required suturing to ensure watertight closure. No clinically significant complications were encountered. CONCLUSION: The modified sutureless sclerotomy technique was found to be safe, more convenient, and easier to perform, especially in eyes with small interpalpebral space. PMID- 10372892 TI - Uveitis associated with varicella virus vaccine. AB - PURPOSE: To report a case of uveitis associated with the live attenuated varicella virus vaccine (Varivax; Merck & Co, Inc, West Point, Pennsylvania) in a young, otherwise healthy girl. METHODS: The time of onset of uveitis in relation to vaccination and the number and the pattern of distribution of vesicles were noted. The patient received oral acyclovir and topical steroids and cycloplegic drops. RESULTS: The uveitis and vesicular rash improved significantly after 7 days of treatment. A literature review and communications with the drug's manufacturer disclosed no identifiable previous cases of uveitis associated with Varivax. CONCLUSIONS: Uveitis should be recognized as a possible adverse side effect of the varicella vaccine. PMID- 10372893 TI - Acute dacryocystitis as a presenting sign of pediatric leukemia. AB - PURPOSE: To report acute dacryocystitis with preseptal cellulitis as the presenting sign of leukemia in a child. METHODS: Case report and literature review. RESULTS: During the initial evaluation of a 17-month-old child with epiphora, left lower eyelid swelling, and a tender left medial canthal mass, a complete blood cell count demonstrated pancytopenia. Bone marrow biopsy disclosed replacement of normal cellular architecture with a dense infiltrate of leukocyte blast forms. DNA analysis disclosed a translocation between chromosome 10 and 11, consistent with the diagnosis of nonlymphocytic leukemia. Although the adjacent lower eyelid cellulitis responded to intravenous antibiotics, lacrimal sac distention decreased only after chemotherapy was initiated. CONCLUSIONS: Dacryocystitis with preseptal cellulitis can be a presenting sign of leukemia. This blood malignancy should be considered in patients whose leukocyte counts do not correlate with their clinical presentation. PMID- 10372894 TI - Clinical findings in a patient with spontaneous arteriovenous fistulas of the orbit. AB - PURPOSE: To report clinical and radiologic findings of a patient with spontaneous arteriovenous fistulas of the orbit. METHOD: Case report. RESULTS: A 73-year-old woman was initially examined with a 1-year history of mild proptosis of the right eye. She had no history of trauma. Neuro-ophthalmologic examination disclosed dilatation of conjunctival vessels, increased intraocular pressure, mild proptosis and bruit in the right eye, and ocular signs suggestive of carotid cavernous sinus fistulas or orbital arteriovenous malformations. The patient exhibited dilation of the superior ophthalmic vein in enhanced computed tomography of the orbit. Selective cerebral angiography disclosed communications between branches of both ophthalmic and facial arteries and the superior ophthalmic vein in the orbit. CONCLUSION: Arteriovenous fistulas of the orbit must be considered in the differential diagnosis of carotid-cavernous sinus fistulas and arteriovenous malformations, although they are quite rare. PMID- 10372895 TI - Antithymocyte globulin treatment of orbital Wegener granulomatosis: a follow-up study. AB - PURPOSE: To describe the follow-up of patients with orbital Wegener granulomatosis after antithymocyte globulin treatment. METHODS: Patients with ocular/orbital Wegener granulomatosis refractory to standard treatment were selected for immunotherapy with rabbit antithymocyte globulin intravenously. The specific ocular/orbital symptoms were monitored in patients with a vision threatening form of Wegener granulomatosis or a life-threatening form with ocular symptoms. RESULTS: One patient had a complete remission, two patients had remissions but still needed additional treatment, and one patient remained refractory. CONCLUSIONS: In selected patients, antithymocyte globulin immunotherapy may present an alternative when vision or life is threatened by orbital Wegener granulomatosis. PMID- 10372896 TI - Peripheral retinal cryotherapy for postvitrectomy diabetic vitreous hemorrhage in phakic patients. PMID- 10372897 TI - Comparison of pneumatic retinopexy and scleral buckling in the management of primary rhegmatogenous retinal detachment. PMID- 10372898 TI - The role of botulinus toxin type A in treatment--with special reference to children. AB - Although botulinum toxin A was first introduced to treat strabismus and blepherospasm it is now used in an increasing number of conditions, many in the field of pediatrics. Its action results from a prevention of the release of acetylcholine from nerve terminals. A number of studies recording the effects of the toxin in the treatment of spastic cerebral palsy are reviewed, and although these can be criticized, there seems to be no doubt that it can be of benefit. It is few side effects, but it may reveal an underlying weakness. Other disadvantages are its cost and the need for repeated injections. It can be used for the relief of rigidity, although the effects in the extrapyramidal form of cerebral palsy are not so dramatic. Also it can be beneficial in some forms of dystonia, rarely if this is generalized, but certainly if it is focal, and especially if there is accompanying pain. There are several conditions seen in children, such as strabismus, blepherospasm and tremors, in which this form of treatment will rarely be indicated; but they will be mentioned. An exception may be spasmodic torticollis during adolescence if it does not respond to other therapy, as it is so disabling. Botulinum toxin can be used to block the discharges from cholinergic sympathetic and parasympathetic terminals. Focal hyperhidrosis can be very distressing among older children, and the use of the toxin should sometimes be considered in this and other autonomic disorders. PMID- 10372899 TI - Appropriate investigations for clinical care versus research in children with autism. AB - As disorders on the autistic spectrum are behaviorally defined, there is no medical test to diagnose autism. The purpose of a medical evaluation is to detect particular etiologies, and manifestations like clinical or subclinical epilepsy or behavior problems that might mandate pharmacologic intervention. Defining a unique syndrome or genetic etiology may benefit other family members, although, currently, specific causes are detectable in only a small minority of individuals on the autistic spectrum. The paper lists elements of the history, examination, and laboratory testing most likely to be informative in clinical practice. Ordering large numbers of tests in the absence of a specific clinical indication is not recommended because it is invasive, wasteful and unlikely to generate useful data. This is not true, of course, in the context of a hypothesis-driven, approved research protocol where collecting standardized data and applying the most up-to-date research technologies is appropriate. PMID- 10372900 TI - Treatment of infantile spasms with zonisamide. AB - We determined the efficacy of and tolerability to zonisamide (ZNS) in newly diagnosed patients with infantile spasms. ZNS, 4-20 mg/kg per day, was introduced as an add-on therapy or monotherapy in 27 children with infantile spasms (cryptogenic, 2; symptomatic, 25). The dosage was initially 2-4 mg/kg per day, and then was increased by 2-5 mg/kg every 2-4 days until the seizures disappeared. Nine (33.3%) out of the 27 patients who were administered ZNS exhibited the disappearance of seizures. ZNS was effective in all the cryptogenic cases and seven (28.0%) of the symptomatic cases. The effective daily doses were 5-12.5 mg/kg (mean, 7.8 mg/kg), and the daily dosages were 40-100 mg (mean, 61.1 mg). The steady-state plasma ZNS concentrations were almost within the therapeutic range. The mean time interval between the start of ZNS therapy and the seizure disappearance was 5.0 days. Six (75.0%) of eight effective cases, the exception being one for whom EEG was not performed after the therapy, showed the disappearance of hypsarrhythmia. The recurrence of seizures were observed in four of the nine cases. No adverse reaction to ZNS was noted in any patient. In conclusion, ZNS treatment is considered to be worthwhile trying, for early stage therapy for infantile spasms. PMID- 10372901 TI - Functional recovery in hemiplegic cerebral palsy: ipsilateral electromyographic responses to focal transcranial magnetic stimulation. AB - The patterns of functional recovery after unilateral cerebral damage occurring in the prenatal to infantile periods were studied in nine patients with hemiplegic cerebral palsy. Motor evoked potentials (MEPs) recorded from the small hand muscles were investigated using focal transcranial magnetic stimulation (TMS). The MEPs findings could be separated into three subtypes based on the features of ipsilateral MEPs elicited by TMS over the unaffected motor cortex. Bilateral MEPs of similar latency were obtained in three patients. These patients each having a congenital lesion invariably exhibited mirror movements and severe hemiparesis. Meanwhile, ipsilateral MEPs with markedly prolonged latency were demonstrated in two other patients, who exhibited synergistic associated movements and severe hemiparesis caused by an acquired lesion. In the remaining four patients, who showed mild hemiparesis without such abnormal interlimb coordinations, there were no ipsilateral MEPs. Thus, we suggest that TMS is useful for confirming the electrophysiological findings relevant to functional recovery in hemiplegic cerebral palsy underlying such abnormal interlimb coordinations. Specifically, bilateral MEPs of similar latency were considered consistent with compensatory mirror movements originating from bilateral motor representation in the unaffected motor cortex. PMID- 10372902 TI - Incidence of childhood epilepsy in Estonia. AB - The aim of this study has been to establish the incidence rate (IR) and main characteristics of childhood epilepsy in Estonia. A population-based prospective study was performed from January 1st 1995 to December 31st 1997 in seven counties (population of children 161202). Only cases occurring in the age range of 1 month to 19 years with active epilepsy were included. Two hundred and sixteen cases met the study criteria. The total age-adjusted IR was 45/100000. The IR was the highest, 73/100000, in the age group from 1 month to 4 years. The IR declined markedly after the age of 15 years. Primarily generalized seizures demonstrated a higher IR, 25/100000, than partial seizures, the IR of which was 20/100000. The IR of symptomatic epileptic syndromes was 16/100000, that of cryptogenic, 15.5/100000 and that of idiopathic, 13/100000. The cumulative incidence of epilepsy through age 19 was 0.13%. A family history of epilepsy was present in 13.9% of cases. In 40.7% of cases the cause of epilepsy was identified. Adverse perinatal events were the most frequent etiological factors: in 25%, IR 11/100000. In 103 cases (47.6%) additional medical problems were disclosed. Strong negative univariate association was noted between partial seizures and idiopathic etiology (OR 0.37, 95%CI 0.18, 0.72; P = 0.002) and between partial seizures and motor disability (OR 0.43, 95%CI 0.24, 0.78; P = 0.003). The incidence of childhood epilepsy in Estonia was comparable with developed countries. Generalized seizures predominated. Perinatal factors were the main causes. The idiopathic etiology and motor disability of cryptogenic and symptomatic cases were associated with generalized seizures. PMID- 10372903 TI - Rett syndrome and genetic drift. AB - An X chromosome gene is assumed to be responsible for the cause of Rett syndrome (RS). However, new genealogical observations suggest involvement of autosomal recessive gene(s) as well, at least in familial cases. To account for these and other recent observations, the theoretical model presented in 1990 by the authors of this paper is applied to the calculation of gene frequencies. Observed frequencies of sporadic and familial cases of RS are used, taking into account genetic drift in inbreeded areas. Moreover, an attempt is made to use the proportion of RS variants in familial and sporadic cases for the explanation of so called 'formes frustes', and as evidence for the existence of female as well as male carriers. The estimated frequency of the recessive autosome mutation, or possibly a frequent polymorphism, is 22.5%. PMID- 10372904 TI - Comparison between semiquantitative interictal Tc-99m HMPAO SPECT and clinical parameters in children with partial seizures. AB - The aim of the present study was to correlate between clinical parameters (age, age of onset, frequency and duration of seizures) and semiquantitative interictal SPECT parameters in children with partial seizures. We obtained 30 patients who had hypoperfusion in interictal SPECT, retrospectively. All patients underwent a detailed clinical examination, electroencephalography (EEG) investigation and brain computerized tomography (CT) and/or magnetic resonance imaging (MRI). Single photon emission computerized tomography (SPECT) studies were evaluated visually and by calculating semiquantitative parameters (the degree (asymmetry index, AI) and extent (number of ROI) of hypoperfusion). Visual analysis detected ipsilateral hypoperfusion in 23 (76%) patients with a unilateral focus and contralateral hypoperfusion in seven patients. We found an inverse correlation between the age at onset of seizure (r = -0.40, P = 0.025), frequency of seizures(but positive correlation; r = 0.77, P = 0.000) and AI. Number of ROIs showed a moderate correlation with the frequency of seizures (r = 0.67, P = 0.000), while correlation of the age at onset of seizures was not significant. This study performed in pediatric patients also suggested that either SPECT parameters may be used for correlating with clinical parameters. PMID- 10372905 TI - Cellular senescence of angiofibroma stroma cells from patients with tuberous sclerosis. AB - Tuberous sclerosis complex (TSC) is an autosomal dominant disorder characterized by epilepsy, mental retardation and hamartomatous lesions in multiple organs. It has been shown that the genes responsible for TSC, TSC1 and TSC2, act as tumor suppressors, but the mechanism of hamartomatous growth in several tissues is not completely understood. The TSC hamartomas are essentially benign and they rarely progress to malignant tumors. In this report, we cultured the angiofibroma stroma cells of three adult TSC patients and compared these cells with normal skin fibroblasts for their proliferative capacity, cell morphology and mitotic cycle using a stain for microtubules and the expression of the senescent associated beta-galactosidase (SA beta-Gal). Cultured angiofibroma stroma cells from TSC patients displayed several characteristics observed in human senescent fibroblasts; a low proliferative capacity, an increase in cell size, increased binucleated cells in association with abnormal cytokinesis and increased SA beta Gal positives. Growth of facial angiofibromas in TSC may be caused by a gain in enhanced sensitivity toward some of the potential mitogens and forced multiplication without loss of the cellular senescent program; this may be the reason why TSC hamartomas rarely progress to malignancy and why the growths are limited to a finite size. PMID- 10372906 TI - Beta-2-microglobulin and ferritin in cerebrospinal fluid for evaluation of patients with meningitis of different etiologies. AB - To determine whether or not the beta-2-microglobulin (beta2-m) and/or ferritin levels in cerebrospinal fluid (CSF) can be used as markers for the differential diagnosis of meningitis and determination of the response to treatment, 122 subjects with etiologically well-characterized diagnoses were classified into three groups: bacterial meningitis (n = 5; mean age +/- SD. 1.0+/-1.0 year), viral meningitis (n = 39; 5.9+/-3.8 years), and a non-meningitis group (n = 78; 5.2+/-4.9 years). The levels of beta2-m and ferritin in CSF were determined by means of a latex photometric immunoassay. The statistical significance of the data was analyzed with the Mann Whitney U-test. A receiver operating characteristic curve was used to evaluate the diagnostic accuracy of each prediction marker. This study indicated that (1) the levels of beta2-m and ferritin in CSF were related with age in the non-meningitis group: subjects of up to 5 months of age exhibited higher concentrations of these proteins than ones of above 6 months of age (beta2-m, 1.89+/-1.13 vs. 0.84+/-0.65 mg/l. P < 0.01; ferritin, 2.97+/-2.04 vs. 1.81+/-1.34 microg/l, P = 0.09); (2) the beta2-m level was significantly higher in the CSF of patients with viral meningitis than in ones without meningitis (2.41+/-1.23 vs. 0.84+/-0.65 mg/l, P < 0.01): the best cut-off value was 1.2 mg/l (3) the ferritin level was significantly higher in the CSF of patients with bacterial meningitis than in ones with viral meningitis (43.24+/-39.49 vs. 6.81+/-7.41 microg/l, P < (.01): the best cut-off value was 7.5 microg/l; and (4) sequential measurement of the CSF ferritin level was of value for determination of the response to antibiotic treatment for bacterial meningitis. These results only apply to patients of greater than 6 months of age. beta2-m and ferritin in the CSF can be used as an ancillary tool for diagnostic guidance in the acute phase of meningitis and determination of the response to treatment for bacterial meningitis. PMID- 10372907 TI - Proton magnetic resonance spectroscopy to study the metabolic changes in the brain of a patient with Leigh syndrome. AB - Localized proton magnetic resonance spectroscopy (MRS) was performed to study the metabolic changes in the brain of a patient with Leigh syndrome, who had a T-->G point mutation at nt 8993 of mitochondrial DNA. In this patient, sodium dichloroacetate therapy normalized the lactate and pyruvate levels in both blood and cerebrospinal fluid (CSF). However, his psychomotor retardation did not improve and magnetic resonance imaging showed progressive cerebral atrophy. In the patient's spectra, elevation of brain lactate was observed throughout the brain with regional variations, predominantly in the basal ganglia and brainstem with an abnormal MRI appearance. Although the lactate/creatine ratio observed on proton-MRS was related to the CSF lactate level, the ratio did not completely parallel the CSF lactate level, i.e. brain lactate was detected even when the CSF lactate level had become normalized. Furthermore, proton-MRS revealed a decrease in the N-acetylaspartate/creatine ratio and an increase in the choline/creatine ratio, representing neuronal loss and breakdown of membrane phospholipids. The clinical and MRI findings were well related to the changes in spectroscopically determined brain metabolites. These results indicate that the brain metabolites observed on proton-MRS are useful indicators of a response to therapy and prognosis in Leigh syndrome. PMID- 10372908 TI - Protracted course of N-acetylaspartic aciduria in two non-Jewish siblings: identical clinical and magnetic resonance imaging findings. AB - Canavan disease (CD) or N-acetylaspartic aciduria (NAA) is a severe, progressive, autosomal recessive leukodystrophy, occurring mainly among Ashkenazi Jewish individuals. We report clinical and MRI findings in two, non-Jewish, Greek siblings, 7 and 5 years, respectively, with a protracted form of NAA. The constellation of identical clinical course and identical MRI findings with involvement of the basal ganglia, the brainstem, the dentate nucleus and the subcortical white matter in both siblings, as well as the absence of the three commonest mutations found in both Jewish and non-Jewish CD patients, give support to the existence of a protracted form of NAA with a milder clinical course, presumably genetically determined. PMID- 10372909 TI - A case of epileptic negative myoclonus: therapeutic considerations. AB - This study presents a patient with epileptic negative myoclonus who showed interictal focal epileptic discharges in the centrotemporal region. The patient's seizures were exacerbated by carbamazepine, zonisamide, and valproate, but completely controlled by ethosuximide, and were suggested to have some relation with thalamocortical oscillation mechanisms. Ethosuximide is supposed to be a drug of worth to try to use in epileptic negative myoclonus patients with centrotemporal spike foci. PMID- 10372910 TI - Convulsive syncope following placement of sphenoidal electrodes. AB - Two cases of convulsive syncope following the insertion of sphenoidal electrodes are reported. The episodes occurred shortly after an uneventful insertion of the needle. Both patients exhibited behavioral arrest with loss of muscle tone, followed by flexor posturing, jerking of the extremities, then followed by what appeared to be a panic attack. Episodes were clinically distinct from the patients' typical spells and were initially interpreted as representing psychogenic events. EEGs during the episodes showed diffuse slowing followed by generalized suppression of rhythms. Simultaneous EKG showed bradycardia followed by brief asystole and then resumption of normal heart rhythms in both cases. Vagally mediated cardioinhibitory reactions induced by fear, pain and possibly stimulation of branches of the trigeminal nerve in the face represent an uncommon but potentially serious complication of placement of sphenoidal electrodes. PMID- 10372911 TI - Moro reflex profile in high-risk infants at the first year of life. PMID- 10372912 TI - Dehydration of 3-hydroxyacyl-CoA in brain very-long-chain fatty acid synthesis. AB - Rat brain microsomes actively dehydrate 3-hydroxyacyl-CoAs. Using chemically synthesized [1-(14)C] (R,S) 3-hydroxyeicosanoyl-CoA, we investigated the biochemical characteristics of the dehydration and reduction steps of stearoyl CoA elongation. The reaction products, separated and identified as trans2,3-enoyl CoAs and, in the presence of NADPH, as saturated acyl-CoAs, were released from the enzyme as thioesters which were partly hydrolysed. A kinetic analysis of the two coupled reactions showed that the 3-hydroxyacyl-CoA dehydrase catalysed a reversible reaction with kinetic constants of about 0.045 min(-1) for forward reaction (dehydration) and 0.025 min(-1) for reverse reaction (hydration); Vmax of the dehydration reached 20 nmoles/min/mg and the apparent Km was 44 microM. In the presence of NADPH, the kinetic constants for the dehydrase were unchanged and that for the trans2,3-enoyl-CoA reductase was 0.025 min(-1). The relative proportion of trans2,3-enoyl-CoA and saturated acyl-CoA depended on the protein amount. An inhibition of the reduction step was observed for substrate concentrations above 15 microM. The 3-hydroxyacyl-CoA dehydrase used (R) rather than (S) 3-hydroxyacyl-CoA. Furthermore, the elongation of (R) 3 hydroxyeicosanoyl-CoA yielded saturated very-long-chain acyl-CoA. These results demonstrated that 3-hydroxyacyl-CoAs entered the elongating complex exclusively at the level of the dehydrase and not of the condensing enzyme. PMID- 10372913 TI - Differential effects of acute and chronic treatment with typical and atypical neuroleptics on c-fos mRNA expression in rat forebrain regions using non radioactive in situ hybridization. AB - The regional difference in the expression of c-fos mRNA in rat forebrain after either acute or chronic administration of typical (haloperidol and fluphenazine) and atypical neuroleptics (clozapine and (+/-)-sulpiride) was investigated. Rats were injected intraperitoneally with vehicle or neuroleptics daily for 14 days. Twenty-four hours after the last injection, the rats were challenged with vehicle or neuroleptics. C-fos mRNA expression was determined by non-radioactive in situ hybridization. Acute treatment with typical neuroleptics induced a remarkable induction of c-fos mRNA in the dorsolateral striatum, whereas this induction was greatly attenuated by chronic administration. All neuroleptics examined induced c fos mRNA in the shell region of N. accumbens by acute administration and this expression was still elevated after chronic treatment. Since chronic neuroleptics do not induce tolerance to their antipsychotic activities, our study suggests that the shell region of N. accumbens is an important target site for antipsychotic effects of neuroleptics. PMID- 10372915 TI - The oxidation-reduction state of serum proteins in multiple sclerosis patients: effect of interferon beta-1b. AB - The concentration of reduction equivalents in serum was studied in a cohort of healthy individuals, in a group of multiple sclerosis (MS) patients undergoing treatment with interferon beta-1b and another group of MS patients who refused treatment with interferon beta-1b. Two classes of sulfhydryl groups were detectable in serum: (1) the uncovered sulfhydryls, accessible to the oxidation reduction substrate 5,5-dithiobis-(-2-nitrobenzoic acid) (DTNB); and (2) the hidden sulfhydryls that required previous heat denaturation of serum proteins to become accessible to DTNB. The concentration of the reduced form of both the uncovered- and hidden-type of sulfhydryls was higher in the serum of MS patients than in healthy individuals. Interferon beta-1b lowered the plasma concentration of the uncovered reduced sulfhydryls after 3 months of treatment. This was in contrast to a minor effect of interferon beta-1b in the hidden-form of sulfhydryl groups. The results suggest that the concentration of reduced sulfhydryls is a biochemical marker of the in vivo oxidation/reduction reactions in MS. PMID- 10372914 TI - Gene expression in developing rat hippocampal pyramidal neurons appears independent of mossy fiber innervation. AB - In the rat, neonatal gamma-irradiation of the hippocampus induces a selective destruction of dentate granule cells and prevents the development of the mossy fiber-CA3 pyramidal cell connection. In the absence of mossy fiber input, the CA3 pyramidal neurons exhibit morphological alterations and rats deprived of dentate granule cells fail to develop kainate-induced epileptic activity in the CA3 pyramidal neurons. Neonatal elimination of the granule cells also impairs learning and memory tasks in adult rats. In the present work, we assessed by in situ hybridization and semi-quantitative RT-PCR, whether in the pyramidal layers, the absence of mossy fiber input alters the expression of a number of genes involved in activity-dependent signal transduction, in GABAergic neurotransmitter signaling and in neurite development via microtubule organization. Surprisingly, we show that the expression and the developmentally regulated alternative splicing of the genes we examined in the developing hippocampus are not altered in the pyramidal neurons, whether the dentate granule afferents are present or absent. Our results suggest that in the CA3 pyramidal layer, the developmental expression patterns of the mRNAs we studied are independent of extrinsic cues provided by mossy fiber input. PMID- 10372916 TI - Na+, K+ and Ca2+ antagonize the glutamate- and glycine-induced decrease of [3H]MK 801 binding observed in the presence of Mg2+ at low pH. AB - NMDA receptors are glutamate-regulated ion channels that are of great importance for many physiological and pathophysiological conditions in the mammalian central nervous system. We have previously shown that, at low pH, glutamate decreases binding of the open-channel blocker [3H](+)-5-methyl-10,11-dihydro-5H dibenzo[a,d]cyclohepten, 5,10-imine ([3H]MK-801) to NMDA receptors in the presence of 1 mM Mg2+ but not in Krebs buffer. Here, we investigated which cations that block the glutamate-induced decrease in Krebs buffer, using [3H]MK 801 binding assays in membrane preparations from the rat cerebral cortex. At pH 6.0, Na+, K+, and Ca2+ antagonized the glutamate-induced decrease with cross-over values, which is a measure of the antagonist potencies of the cations, of 81, 71, and 26 mM, respectively, in the absence of added glycine. Thus, in Krebs buffer only the concentration of Na+ (126 mM) is sufficiently high to block the glutamate-induced decrease observed at low pH. In the presence of 1 mM Mg2+ and 10 mM Ca2+ at pH 7.4, the cross-over values for Na+, K+, and Ca2+ were 264, 139, and 122 mM, respectively, in the absence of added glycine. This is the same rank order of potency as observed at pH 6.0, suggesting that the less H+-sensitive and the less Ca2+-sensitive, glutamate-induced decreases in [3H]MK-801 binding represent the same entity. The glycine site antagonists 7-chlorokynurenate (10 microM) and 7-chloro-4-hydroxy-3-(3-phenoxy)phenyl-2(H)-quinoline (L-701,324; 1 microM) antagonized the glutamate-induced decrease in [3H]MK-801 binding observed in presence of Mg2+ at pH 6.0, suggesting that glycine is required together with glutamate to induce the decrease observed at low pH. These results suggest that in addition to a previously described high-affinity binding site for H+ and Ca2+ there exist a low-affinity binding site for H+, Ca2+, Na+, and K+ on NMDA receptors. The latter site may under physiological conditions be blocked by Na+ or K+, depending on the extra/intracellular localization of the modulatory site. Both the high-affinity and low-affinity cation sites mediate antagonistic effects on the glutamate- and glycine-induced decrease of the affinity of the [3H]MK-801 binding site, which may correspond to similar changes in the affinity of the voltage-sensitive Mg2+-block site inside the NMDA receptor channel pore, which in turn may affect current and Ca2+ influx through activated NMDA receptor channels. PMID- 10372917 TI - ATPalphaS is a ligand for P2Y receptors in synaptosomal membranes: solubilization of [35S]ATPalphaS binding proteins associated with G-proteins. AB - ATPalphaS was established as a P2Y receptor-specific ligand for assaying the solubilization of functional native P2Y receptors from synaptosomal membranes. These receptors are not yet amenable to biochemical studies. High-affinity [35S]ATPalphaS binding sites in synaptosomal membranes, solubilized with Brij58, retained the binding affinity and ligand specificity (ATPalphaS = ATP > 2-MeSATP > ADP, ADPbetaS > AMP >>> alpha,beta-MeATP) corresponding to P2Y receptors. Mg2+ but not Ca2+, enhanced high-affinity [35S]ATPalphaS binding 30-fold, supporting specific recognition by P2Y receptors. ATPalphaS stimulated P2Y receptor-mediated [35S]GTPgammaS binding equipotently with ATP in synaptosomal membranes and in Brij58-solubilized proteins demonstrating the association with G-proteins. Anion exchange chromatography of solubilized synaptosomal membrane proteins yielded two fractions in which [35S]ATPalphaS binding was regulated by GTPgammaS/Mg2+, thus possibly by heterotrimeric G-proteins. After a second chromatographic step (hydroxyapatite) the regulation of high-affinity [35S]ATPalphaS binding by Mg2+ was still present, whereas the regulation by GTPgammaS/Mg2+ was lost indicating the dissociation from G-proteins. Thus, conditions were found to stabilize ligand binding activity of solubilized P2Y receptors and to solubilize P2Y receptors associated with G-proteins. PMID- 10372918 TI - Nicotinic receptors modulating ACh release in rat cortical synaptosomes: role of Ca2+ ions in their function and desensitization. AB - Cholinergic nerve terminals in the central nervous system are endowed with both muscarinic and nicotinic autoreceptors, mediating inhibition, and enhancement of acetylcholine release, respectively. Exogenous acetylcholine inhibited the K+(15 mM)-evoked overflow of [3H]acetylcholine from superfused rat neocortical synaptosomes; however, in the presence of atropine, this muscarinic inhibition was reversed into a nicotinic potentiation when acetylcholine was added concomitantly with high-K+, but not before depolarization. Increasing concentrations of acetylcholine (plus atropine), nicotine and (+)-anatoxin-a produced elevations of the K+-evoked [3H]acetylcholine overflow resulting in bell shaped concentration-response curves. Synaptosomes pretreated with different concentrations (10 microM to 0.001 microM) of acetylcholine or nicotine responded to a subsequent nicotinic stimulus (10 microM acetylcholine plus 0.1 microM atropine, in 15 mM K+) in a manner reflecting varying degrees of desensitization. This desensitization could be reversed by washings with standard medium and desensitization was attenuated when external Ca2+ ([Ca2+]e) was decreased. Lowering of [Ca2+]e or chelation of internal Ca2+ with 1,2-bis(2 aminophenoxy)ethone-N,N,N',N'-tetracetic acid acetoxymethylester (BAPTA-AM) permitted the nicotinic response to acetylcholine alone (no atropine added) to prevail over the muscarinic response. Pretreatment with BAPTA-AM could however not prevent desensitization by acetylcholine (10 or 0.001 microM). The data indicate that Ca2+ ions are involved in determining the balance between muscarinic and nicotinic autoreceptor function and in the desensitization of nicotinic autoreceptors. PMID- 10372919 TI - Ischemia-induced modifications of protein components of rat brain postsynaptic densities. AB - Considering that postsynaptic densities (PSD) are a functionally active zone involved in excitatory synaptic transmission we evaluated the influence of global, postdecapitative cerebral ischemia of 15 min duration on characteristic protein constituents of PSD in rats. Ischemia induced changes in the assembly and function of calcium, calmodulin-dependent kinase II (CaMKII), calpains and a novel, 85 kDa/RING3 kinase but to different extents. CaMKII is translocated toward the PSD very rapidly and extensively after the first seconds of ischemia. Concomitantly, the total phosphorylating potency of this kinase with endogenous, as well as exogenous, substrates was elevated but to a lower extent than suggested by the increased protein content. Of the two brain-specific isoforms of calpain (mu and m), only recently recognized in PSD, the proteolytically activated, 76 kDa subunit of mu-calpain was significantly down-regulated after 15 min of brain ischemia. However, this effect is coupled with the decline of fodrin, the only calpain substrate that has been demonstrated to be a calpain target in vivo. Together, these findings may suggest that calpains, primarily activated by calcium in ischemic PSD, are subsequently degraded. A new observation is the relatively high phosphorylating activity of a novel, 85 kDa/RING3 kinase in the PSD which independently of other kinase systems, was greatly enhanced after ischemia. These data provide evidence that the signal transduction processes could be rapidly altered by short-term (15 min) brain ischemia due to changes in the assembly and function of PSD connected proteins. PMID- 10372920 TI - Activation of MAPK by potassium bisperoxo(1,10-phenanthroline)oxovanadate (V). AB - Potassium bisperoxo(1,10-phenantroline)oxovanadate (V) [bpV(phen)] is a potent protein tyrocine phosphatase inhibitor which mediates a variety of biological effects. The aim of these studies was to examine the role(s) of mitogen activated protein kinase (MAPK) pathways in PC12 cell proliferation and toxicity by bpV(phen). BpV(phen) exerts a bimodal effect in PC12 cells: proliferation at low and cell death at higher micromolar concentrations. Activation of MAPK by bpV(phen) depends on time and concentration. The phosphorylation pattern of extracellular regulated kinases (ERK 1/2), c-jun N-terminal activated kinases (JNK) and p38 in PC12 cells is strikingly different. Activation of JNK is sustained in PC12 cells. In contrast, ERK 1/2 activation is transient and treatment with PD98059 indicates that ERK activation by bpV(phen) is partly independent from the ras-MEK pathway. Stability studies of bpV(phen) in DMEM and PBS showed linear relationship with T1/2 about 6 h and 10 days in DMEM and PBS, respectively. Comparison between the time courses of MAPK activation and kinetics of bpV(phen) decomposition as assessed by 51V-NMR analysis show that the initial and maximal phosphorylation signals are produced in the presence of the complex bpV(phen) and not caused by the decomposition products of bpV(phen). PMID- 10372921 TI - A possible role of nitric oxide in the regulation of dopamine transporter function in the striatum. AB - Brain microdialysis and high-performance liquid chromatography with electrochemical detection were used to study the effect of the nitric oxide synthase inhibitor Nomega-nitro-L-arginine methyl ester (L-NAME) on striatal dopamine (DA) release in the anesthetized rat. Systemic administration of L-NAME (10 mg/kg, i.p.) significantly decreased the resting release of DA. The peak effect (23% decrease) was reached 45 min after injection. The inactive enantiomer D-NAME (10 mg/kg, i.p.) or the vehicle (saline, 5 ml/kg i.p.) had no effect on the striatal DA level. Neither treatment altered significantly the concentration of dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA). To investigate the possible involvement of the DA uptake system L-NAME was injected also in the presence of the DA uptake inhibitor nomifensine. Local application of nomifensine (10 microM in the dialysate medium) increased the extracellular concentration of DA to about eight-fold of the basal value and stabilized it at this higher level. Under these conditions L-NAME (10 mg/kg, i.p.) was not able to alter the striatal DA level. Neither nomifensine nor L-NAME caused any change in the level of DOPAC and HVA. Our data suggest that endogenously produced nitric oxide may influence the activity of the DA transporter which effect may have special importance in the regulation of extracellular transmitter concentration in the striatum. PMID- 10372922 TI - Hemolytic anemia and the treatment of chronic hepatitis C. PMID- 10372923 TI - Quality assurance for endoscopic disinfection. PMID- 10372924 TI - Endoscopic disinfection: a worldwide problem. AB - Safety of endoscopic procedures has been a major issue over the last 10 years. Outbreak of new infectious diseases (type C) hepatitis, Creutzfeldt-Jakob disease) underlines the necessity for strengthening cleaning and disinfection guidelines. Patients should be ensured that all endoscopic procedures are carried out with high-level disinfection endoscopes and with sterile or single-use accessories. Improvements from a hygienic point of view of both endoscopes and washer-disinfectors are important goals for manufacturers. Adequate training of endoscopic staff is one of the most crucial points to achieve the highest quality control standards in digestive endoscopy. PMID- 10372925 TI - Pill esophagitis. AB - Nine hundred seventy-nine cases of pill esophagitis due to nearly 100 different medications are reviewed. Pill-induced injuries occur when caustic medicinal pills dissolve in the esophagus rather than passing rapidly into the stomach as intended. Most patients suffer only self-limited pain, but esophageal hemorrhage, stricture, and perforation may occur, and fatal injuries have been reported. The incidence of this iatrogenic injury can be reduced but not eliminated by emphasizing the importance of taking pills while upright and with plenty of fluids. PMID- 10372926 TI - Nutritional management of chronic intestinal pseudo-obstruction. AB - Chronic intestinal pseudo-obstruction (CIP) is a gastrointestinal motility disturbance characterized by recurrent episodes of postprandial nausea and bloating in the absence of mechanical obstruction of the small bowel or colon. Weight loss and severe malnutrition are often seen in advanced stages of the disorder. This article discusses the nutritional management of patients with CIP, focusing on general dietary as well as enternal and parenternal nutritional support. Enteral access methods and various enteral formulas used in CIP are also discussed. PMID- 10372927 TI - Changes in serum hepatitis C virus RNA in interferon nonresponders retreated with interferon plus ribavirin: a preliminary report. AB - Ribavirin, a nucleoside analogue, inhibits replication of RNA and DNA viruses and may control hepatitis C virus (HCV) infection through modulation of anti inflammatory and antiviral actions. Ribavirin monotherapy has no effect on serum HCV RNA levels. In combination with interferon, this agent appears to enhance the efficacy of interferon. The aim of this study was to monitor serum HCV RNA levels early during therapy with interferon and ribavirin compared with that previously seen in the same patients during interferon monotherapy. Five patients who previously showed no response to therapy with interferon alfa 3 MU three times weekly for 6 months were retreated with the identical dose of interferon alfa 2b in combination with oral ribavirin 1,000 mg/day. Serum HCV RNA levels were monitored at baseline, week 4, week 8, and week 12 of therapy by a quantitative multicycle polymerase chain reaction assay. In the first 8 to 12 weeks, serum HCV RNA levels showed a greater decrease in all patients when retreated with combination therapy compared with interferon alone. Mean (+/- SEM) serum HCV RNA levels for interferon therapy alone were 3.3 +/- 0.95, 1.2 +/- 0.95, 1.6 +/- 1.2, and 2.3 +/- 1.2 x 10(6) copies/ml at week 0, 4, 8, and 12, respectively. This was compared with 3.3 +/- 0.83, 0.3 +/- 0.2, 0.03 +/- 0.02, and 0.15 +/- 0.14 x 10(6), respectively, for the interferon and ribavirin group (p < 0.07 at week 8). Two of five patients had undetectable serum HCV RNA during combination therapy. Combination therapy with interferon and ribavirin in prior interferon nonresponders reduces serum HCV RNA levels compared with interferon alone. This may suggest some additional antiviral effect of ribavirin when given with interferon. PMID- 10372928 TI - Hepatitis B vaccination in hospital personnel and medical students. AB - We determined the prevalence of hepatitis B markers and the compliance to hepatitis B vaccination in a University Hospital of Santa Maria, Lisbon. The program was begun in 1989 for all hospital personnel and students of the medical school. The screening included 2,360 health care workers and 1,153 students; 57% (2,360/4,103) of hospital personnel and 41% (1,153/2,779) of medical students appeared for vaccination. The prevalence of hepatitis B markers was 16.8% (397/2,360) for hospital personnel and 5.5% (64/1,153) for students, the chronic carrier appearing in 0.95% (22/2,360) of hospital personnel and 0.3% (4/1.153) of students. The departments with the highest prevalence were the Biochemical Laboratory (64%, 7/11), Surgery (42%, 13/31), Pulmonary (39%, 9/23), Emergency (29%, 7/24), Hematology Laboratory (29%, 7/24), and Orthopedics (29%, 10/35). The prevalence was higher in students in the last 3 years of medical school than those in the first 3 years (12.2% [22/181] vs. 7.2% [8/110], p = NS). Adverse effects to vaccination occurred in 14.5%, with local pain the most frequent in 8.6%. The serologic efficacy was 95% (1,044/ 1,097). A nonresponse was observed in male workers, 13% (26/200) compared with 5% (45/897) for females (p < 0.05). Older employees also showed higher nonresponse: The average age of workers with anti-HBs of 0 IU/l was 52.3 years and those with anti-HBs of more than 100 IU/l was 38.4 years (p < 0.02). Hepatitis B vaccination is safe and effective. Our study shows the need for a more aggressive approach to the vaccination of health care workers because a significant percentage of them are not protected. PMID- 10372929 TI - Spontaneous intrahepatic hemorrhage and hepatic rupture in the HELLP syndrome: four cases and a review. AB - Subcapsular hemorrhage and hepatic rupture are unusual catastrophic complications of the HELLP (hemolysis, elevated liver enzymes, and low platelets) syndrome. A high index of suspicion and prompt recognition are keys to proper diagnosis and management of affected patients. The optimal management of these patients is evolving. An aggressive multidisciplinary approach has considerably improved the morbidity and mortality associated with these complications. We present our experience with four cases of hepatic hemorrhage occurring in association with the HELLP syndrome and review the literature on this subject. All of our patients were multiparous, and three had a history of eclampsia/preeclampsia in a previous pregnancy. All four patients developed intrahepatic hemorrhage; two developed hepatic rupture requiring surgical intervention. Three patients developed disseminated intravascular coagulation and acute renal failure. Two patients developed pericardial effusion, pleural effusions, and ascites. One patient died of septic complications after multiple surgical interventions. PMID- 10372930 TI - Utility of technetium-99m-labeled-galactosyl human serum albumin scintigraphy for estimating the hepatic functional reserve. AB - Technetium-99m-diethylenetriaminepentaacetic acid-galactosyl-human serum albumin (Tc-GSA) is a receptor binding agent, specific for asialoglycoprotein receptor, that resides exclusively on the plasma membrane of mammalian hepatocytes. The usefulness of Tc-GSA for estimating the hepatic functional reserve was retrospectively evaluated in patients undergoing a hepatic resection. Tc-GSA scintigraphy was performed in 35 patients before hepatectomy, and the hepatic uptake ratio (LHL15) was calculated. The LHL15 was then compared with the findings of conventional liver function tests, the indocyanine green retention rate in 15 minutes (ICG R15), and histologic activity index (HAI) score. Significant correlations were observed between the LHL15 and values of ICG R15, prothrombin time activity, serum levels of total bilirubin, hyaluronic acid, and values of HAI score. Ratios of LHL15 to preoperative liver volume (LHL-V) correlated well with the regenerative rates of the residual liver after major hepatectomy. In addition, patients with more than 0.76 of LHL-V value had no complications in postoperative course, whereas those with less than 0.73 had several complications due to hepatic dysfunction. Tc-GSA scintigraphy thus appears to be a useful diagnostic tool for evaluating functioning mass of the liver and the values of LHL-V seems to be able to demonstrate regenerative activity in the residual liver after hepatectomy. PMID- 10372931 TI - Transcatheter arterial chemoembolization for hepatocellular carcinoma in patients with Child's grade A or B cirrhosis: a multivariate analysis of prognostic factors. AB - We evaluated factors affecting long-term survival after transcatheter arterial chemoembolization (TACE) for hepatocellular carcinoma (HCC) complicating cirrhosis. One hundred eighty-two patients with Child's class A or B cirrhosis and an HCC, not amenable to surgery or percutaneous ethanol injection, underwent 346 TACEs (mean 1.9) with epirubicin, iodized oil, and gelatin sponge. Many prognostic factors were subjected to univariate analysis and thereafter, when significant, to the Cox's hazard proportional model. Finally, the significant indices in the Cox's model were used to estimate the accuracy of the probability of death with computation of the area under the receiving operative characteristic (ROC) curve. The cumulative survival rates at 1, 2, 3, and 5 years were 0.83, 0.52, 0.40, and 0.16, respectively. According to Cox's model, the factors associated with significantly worse survival were the presence of ascites (p = 0.0027), elevated bilirubin levels (p = 0.0163), elevated alpha-fetoprotein (alphaFP) values (p = 0.0067), a tumor greater than 5 cm in diameter (p = 0.0001), and the absence of a tumor capsule-like rim (p = 0.0278). According to these parameters, the accuracy of the probability of death estimated with ROC analysis was 0.63. Minor and major complications occurred in 82 patients (45%) and caused death in 2 patients. Long-term prognosis after TACE for HCCs in patients with Child's class A or B cirrhosis depends on the presence of ascites, the bilirubin level, the alphaFP value, the diameter of the tumor, and the presence of a tumor capsule-like rim. However, when considered altogether, these variables are poor predictors to evaluate survival, and other factors should be investigated to identify subjects more responsive to TACE. Complications occur in a high percentage of patients, but they do not affect long-term prognosis. PMID- 10372932 TI - Allergic reactions to 6-mercaptopurine during treatment of inflammatory bowel disease. AB - Hypersensitivity reactions to 6-mercaptopurine (6-MP) or azathioprine occur during the treatment of inflammatory bowel disease (IBD), raising significant diagnostic and therapeutic challenges. Charts of 591 patient with IBD treated with 6-MP in a single center were retrospectively reviewed. All allergic reactions were recorded along with results of rechallenge, desensitization, and subsequent course of IBD. Sixteen (2.7%) allergic reactions to 6-MP were noted, with fever being the most common (14 cases). Nine of these were rechallenged with 6-MP with recurrence of the same symptoms. Azathioprine was tried in six patients and in five the same symptoms recurred. Four patients underwent successful desensitization to either 6-MP or azathioprine; all four plus another patient who tolerated direct switch to azathioprine entered long-term remission. Among the remaining 11, 5 required surgery, 2 are well on methotrexate, and 4 have chronic symptoms while being treated with other medications. If an allergic reaction to 6 MP occurs during the treatment of IBD, direct switching to azathioprine is probably not justified. Instead, desensitization to either 6-MP or azathioprine should be attempted. Patients who can tolerate these medications after previous allergic reactions have improved outcomes compared with patients who resort to other forms of treatment. PMID- 10372933 TI - "Reappearance" of Helicobacter pylori after eradication: implications on duodenal ulcer recurrence: a prospective 6 year study. AB - We estimated the rate of Helicobacter pylori "reappearance" and of duodenal ulcer relapse up to 6 years after eradication of H. pylori. Of 220 patients in whom H. pylori was eradicated, 165 were eligible at 12 months to follow-up. Endoscopy was scheduled every 12 months or whenever symptoms appeared. Baseline H. pylori eradication was confirmed by CLO test, histology (hematoxylin-eosin and Giemsa stain), and culture. H. pylori was tested for by the three methods at 12 months and subsequently by 2 methods (CLO, histology) on biopsies obtained from the gastric antrum and body. We reviewed 90 patients after 1 year, 32 after 2 years, 13 after 3 years, 12 after 4 years, 2 after 5 years, and 16 after 6 years (range, 12 to 72 months; average, 25.23 months; patient-years, 347). At 12 months after eradication, 16 of 165 patients (9.7%) were H. pylori positive and 5 had ulcer relapse. Of 75 patients evaluated at 24 months, 7 (9.3%) were H. pylori positive and 1 (1.3%) had ulcer relapse. At 36 months, 43 patients were seen and 1 (2.3%) was H. pylori positive and had ulcer relapse (2.3%). Thirty, 18, and 16 patients were seen at 48, 60, and 72 months, respectively. None was H. pylori positive and none had ulcer relapse. Overall, 24 H. pylori-positive patients were found, two thirds of them in the first year after eradication. In 7 of 24 (29%, 6 smokers), ulcer recurred. None of the H. pylori-negative patients had ulcer relapse. The H. pylori reappearance rate was 7% and the ulcer relapse rate was 2% per patient year. If the 16 H. pylori-positive patients who were found the first year are considered as recrudescence, then the reinfection rate will be 2.3% per patient year. PMID- 10372934 TI - Helicobacter pylori eradication: comparison of three treatment regimens in India. AB - Conventional bismuth-based triple therapy has multiple problems, such as inadequate drug compliance, side effects, and drug resistance. Combination of omeprazole and clarithromycin with or without combination with antibiotics like amoxycillin has been shown to be effective in eradication of Helicobacter pylori. Reports from India are few on the efficacy of clarithromycin-based drug combinations. Therefore, we evaluated efficacy of omeprazole and clarithromycin with or without amoxycillin for treating H. pylori infection. Sixty-four consecutive patients with upper gastrointestinal symptoms and having H. pylori infection were included. In every patient, complete upper gastrointestinal endoscopy was done. H. pylori infection was diagnosed by identification of organism on antral biopsies and positive rapid urease test. Patients were treated with omeprazole 40 mg/day + clarithromycin 250 mg twice daily (group I, n = 22), or omeprazole 40 mg/day + clarithromycin 250 mg twice daily + amoxycillin 500 mg three times daily (group II, n = 20), or bismuth subcitrate 120 mg four times daily + amoxycillin 500 mg three times daily + metronidazole 400 mg three times daily (group III, n = 22) for 2 weeks. H. pylori status was reevaluated 1 month after completion of treatment. One patient in each group stopped drugs due to side effects. Eradication rate was not significantly different in group I (15/22, 68%), group II (14/20, 70%), and group III (13/22, 59%). Of those completing therapy, side effects were observed in three patients in group III (nausea, skin rash, metallic taste), whereas none of the patients in group I and group II had any side effects. Addition of amoxycillin did not appear to improve efficacy of dual omeprazole and clarithromycin therapy and appeared to be no different than bismuth, metronidazole, and amoxycillin triple therapy. Overall, none of regimens was particularly good. PMID- 10372935 TI - Pressure-overload-induced sliding hiatal hernia in power athletes. AB - Sliding hiatal hernias are a common condition thought to occur with increasing age secondary to a degenerative process. The incidence of sliding hiatal hernias in the general population is 0.5%. Although the prevalence in the Western world is thought to be significantly higher, with approximately 60% of geriatric patients in North America having a hiatal hernia on radiologic studies. Thus, the primary etiology of the sliding hiatal hernia is thought to be degeneration of the phrenoesophageal ligament. Most hiatal hernias occurring in young adults are idiopathic. There has been speculation of a stress-induced hiatal hernia from repeated episodes of elevated intra-abdominal pressure, and to date there is one report of a pressure-overload-induced hiatal hernia occurring in an elite body builder. The prevalence of hiatal hernia in young male power athletes has yet to be examined. Therefore, we examined eight male elite power athletes and seven male non-weightlifters, matched for age, via fluoroscopy with barium swallow to test the hypothesis that pressure overload can induce hiatal hernias in young adults. PMID- 10372936 TI - Relapsing hepatitis A: a case report and review of the literature. PMID- 10372938 TI - Hemolytic anemia secondary to alpha-interferon treatment in a patient with chronic C hepatitis. PMID- 10372937 TI - Double papillary orifice: anatomic variant or choledochoduodenal fistula? PMID- 10372939 TI - Esophageal rupture after regional anesthesia: report of two cases. AB - Esophageal perforation after anesthesia is rare. It is usually secondary to esophageal instrumentation. Only one case of barogenic rupture after regional anesthesia has been reported. We report two additional cases and present possible mechanisms for this unusual entity. Neither patient had anatomic abnormalities by history or preoperative endoscopy. However, both patients and the previously reported patient had esophageal dysmotility resulting from advanced age, alcoholism, intraoperative medications, and preexisting disease. Each patient experienced at least one episode of emesis with subsequent perforation of the distal one third of the esophagus. The previously reported patient died; both of our patients underwent successful surgical repair and are alive 2 years later. Intraoperative or postoperative emesis in patients with esophageal dysmotility appears to be the principal factor causing esophageal rupture after regional anesthesia. Prevention of nausea and vomiting and recognition of this high-risk population may minimize this complication in the future. PMID- 10372940 TI - Autoimmune pancreatitis, pancreatic mass, and lower gastrointestinal bleed. AB - Autoimmune pancreatitis (AIMP) is a recently described clinical entity causing chronic pancreatitis. It often presents with diffuse enlargement of the pancreas and/or a focal mass at the head of the pancreas causing common bile duct obstruction and jaundice. In most instances, AIMP is mistaken for pancreatic cancer. A number of laboratory abnormalities such as positive antinuclear antibody, hypergammaglobulinemia, and antibody to carbonic anhydrase are often present in these patients. Currently, pancreatic biopsy demonstrating characteristic histopathologic changes is essential to establish the diagnosis. We report the first case of AIMP presenting as a pancreatic tail mass and lower gastrointestinal bleed. PMID- 10372941 TI - Eosinophilic enterocolitis and visceral neuropathy with chronic intestinal pseudo obstruction. AB - A patient is described who was found to have both eosinophilic enterocolitis and visceral neuropathy with chronic idiopathic intestinal pseudo-obstruction. The etiology and pathogenesis of this rare combined disorder of the gastrointestinal tract are discussed. Although eosinophilic enterocolitis is amenable to conservative treatment, surgery may be necessary for palliation in selected patients with pseudo-obstruction due to visceral neuropathy. PMID- 10372942 TI - Familial adenomatous polyposis: a case report and histologic mucin study. AB - Adenocarcinoma arising at an ileostomy is uncommon, and only 29 cases have been reported in the literature. The case of a 54-year-old man who developed an adenocarcinoma at a Brooke ileostomy is reported. The ileostomy had been fashioned 21 years earlier after proctocolectomy for familial adenomatous polyposis (FAP). A wide local excision of the stoma was performed, and a new Brooke ileostomy was fashioned on the opposite side of the abdomen. Histopathologic examination revealed a well-differentiated adenocarcinoma with early invasion of the submucosa. On hematoxylin and eosin staining, the ileal mucosa adjacent to the tumor showed signs of colonic metaplasia, including loss of villous architecture and a reduced number of Paneth cells. Mucin staining using the high iron diamine-alcian blue stain demonstrated a mixture of sulfomucin and sialomucin in the ileal mucosa near the tumor, confirming colonic metaplasia. Ileostomy site carcinogenesis can be attributed to both the colonic metaplasia and the inherent nature of FAP or ulcerative colitis (UC), where colonic mucosa is susceptible to adenoma formation or dysplasia. Longstanding ileostomies in patients with FAP or UC should be followed to exclude the development of adenoma, dysplasia, or cancer. PMID- 10372943 TI - Carcinoma in villous adenoma of ascending colon associated with sarcoid reaction in the regional lymph nodes. AB - A 79-year-old woman was admitted to our hospital due to continuous anal bleeding. Colonoscopy showed a huge villous tumor on the middle area of the ascending colon. A typical right colectomy and lymph node dissection were performed. The resected specimen showed a villous type tumor located on the ascending colon. The histopathologic investigation demonstrated a moderately differentiated adenocarcinoma arising in a tubulovillous adenoma and extending to the submucosa. Although there was no evidence of metastatic carcinoma in the dissected lymph nodes, epithelioid cell granulomas with multinucleated giant cells lacking in the central caseous necrosis suggested sarcoid reaction. PMID- 10372944 TI - Ulcerative colitis complicated by dysplasia-adenoma-carcinoma in a man with Bloom's syndrome. AB - Bloom's syndrome (BS) is a rare genetic disorder in which the major clinical feature is growth deficiency. The genome in BS somatic cells is unstable, and hypermutability explains many clinical features. Most notably, affected persons are at enormously increased risk of developing many types of cancers at different sites. It has been well known that ulcerative colitis (UC) is associated with the spectrum of epithelial changes signifying dysplasia and the progression to frank carcinoma. We report here a case of UC complicated by dysplasia-adenoma-carcinoma sequence in a 37-year-old man with BS. PMID- 10372945 TI - Severe jaundice in a gunshot casualty due to the coexistence of Dubin-Johnson and glucose-6-phosphate dehydrogenase deficiency. AB - We report an unusual case of a 21-year-old man who was shot in his abdomen in the course of a robbery. He was previously diagnosed as glucose-6-phosphate dehydrogenase deficient. Mild icterus was noticed on admission to the emergency room. Exploratory laparotomy revealed a perforated ileal loop that was resected, and because the liver color was greenish black, a liver biopsy was performed during the operation. After operation the patient went through a severe icteric state that resolved spontaneously within a few days. Urinary coproporphyrin levels, along with compatible liver biopsy, confirmed the diagnosis of Dubin Johnson disease. Severe hyperbilirubinemia after an abdominal injury is uncommon and is usually due to either a biliary duct injury or iatrogenic injury. This case presents an unusual cause of severe postoperative jaundice due to the rare coexistence of two inherited disorders. PMID- 10372946 TI - Fulminant herpes hepatitis in a healthy adult: a treatable disorder? AB - Hepatitis due to herpes simplex virus (HSV) is a potentially fatal disorder that is often not considered in the differential diagnosis of acute hepatitis. This disease occurs most often in patients with impaired immunity and is very uncommon in healthy patients. HSV hepatitis presents with a wide clinical spectrum, and the clinical diagnosis is difficult. We describe a case of disseminated herpes virus infection with fulminant hepatitis mimicking an acute human immunodeficiency virus infection in a 33-year-old healthy man. Preliminary studies suggest that early treatment of HSV hepatitis with acyclovir may be beneficial in these patients. A high index of suspicion and the availability of early diagnostic tools, such as HSV DNA detection, may dramatically improve the clinical outcome of severe HSV hepatitis. PMID- 10372947 TI - Alzheimer disease ethics: a continuing area of public interest. PMID- 10372948 TI - Ethical issues in Alzheimer disease: the experience of a national Alzheimer society task force. AB - There has been increasing recognition of the ethical dilemmas that arise in the delivery of health care services and in planning and executing scientific research. Alzheimer disease (AD) and related dementias pose a particular challenge for families, care providers, and researchers because of the nature of the illness. Naturally, those at potential risk of developing the disease are eager for scientists to develop valid predictive tests for the disease. Alzheimer organizations have developed worldwide in response to the growing awareness and knowledge of the effects of dementia on individuals and their families. These organizations have played a role in advocating for research, increasing general awareness of the nature of the disease, and lobbying for more services for persons with dementia and their families. These organizations have also realized the increasing concern about the many ethical issues that arise in caring for those with AD and researching causes and cures. This paper describes a unique process one national Alzheimer society used to develop an Ethics Task Force to provide guidelines on ethical issues. PMID- 10372949 TI - Alzheimer's Disease International and International Working Group for Harmonization of Dementia Drug Guidelines for research involving human subjects with dementia. PMID- 10372950 TI - Assessment of mental competency in community-dwelling elderly. AB - We studied the utility of a "vignette method" to assess mental competency for decision making on medical treatment and research participation. A vignette is a description of an imaginary situation in which the subject is asked to decide on a proposed treatment or on participation in research. His or her understanding of the situation and the quality of the reasoning underlying that choice are tested by a short series of questions. Subjects were participants in the Amsterdam Study of the Elderly (AMSTEL), a population-based study on cognitive decline and dementia. The sample consisted of elderly people (70-90 years), who were cognitively intact (n = 176) or had a dementia syndrome (n = 64; mostly Alzheimer disease). Dementia was diagnosed using the Cambridge Examination for Mental Disorders of the Elderly (CAMDEX) schedule. Two vignettes were used as competency assessment instruments. The answers to the vignette questions were summed to form a competency score. The reliability (internal consistency) of this score was 0.82 for both vignettes combined. After dichotomization into competent/incompetent (cutoff at the fifth centile of the control group), the agreement between the vignette method and a physician's judgment of competency was poor (kappa = 0.36) in the demented group. There was no agreement whatsoever when subjects with "minimal dementia" (n = 14) were left out of this analysis (kappa = 0.04). As expected, mean competency scores declined with increasing dementia severity. A multiple regression analysis showed that mental competency as measured by the vignette method was determined mainly by recent memory, expressive language, and abstract thinking. In the control group the competency score was only slightly related to education (r = 0.12) and verbal intelligence (r = 0.27). We conclude that the vignette method is a reliable and valid method for the assessment of mental competency in elderly people with cognitive decline. The vignette method is preferred over a physician's judgment, especially in patients with early dementia. PMID- 10372951 TI - Suicide in two patients with a diagnosis of probable Alzheimer disease. AB - Two patients meeting the criteria for probable Alzheimer disease (AD) who were participating in a phase 3 clinical program with eptastigmine, a cholinesterase inhibitor, committed suicide. The first patient committed suicide by a self inflicted gunshot wound to the head. The second patient committed suicide by jumping from a 19th story window. These two patients shared several clinical features with those found in the literature: being at the early stages of the disease, having a high level education, with preserved insight, having access to firearms, and being aware of not responding to pharmacological treatment. PMID- 10372952 TI - Ethics of end-of-life decisions in cases of dementia: views of the Royal Dutch Medical Association with some critical comments. AB - Some of the views of the Royal Dutch Medical Association on end-of-life decisions in cases of people suffering from Alzheimer disease and related disorders are presented. The focus of the present report is on the views of the commission regarding active life termination of demented patients with or without an actual and explicit request from the patient. Some comments on these views are made, particularly regarding the notion of "loss of human dignity" (ontluistering) with respect to dementia. PMID- 10372953 TI - Perception of emotion in frontotemporal dementia and Alzheimer disease. AB - Frontotemporal dementia (FTD) is the second cause of degenerative dementia. Behavioral changes occur before the cognitive decline and remain the major feature. A poor perception of emotion could account for some behavioral symptoms. The aim of this study was to assess the perception of emotion in patients with FTD and to compare it with that of patients with Alzheimer disease (AD). Fifty subjects performed the tests: 20 patients with probable AD, 18 patients with FTD, and 12 matched controls. The two patient groups did not differ in age, sex, severity of dementia, duration of the disease, and language tests. Subjects had to recognize and point out the name of one of seven basic emotions (anger, disgust, happiness, fear, sadness, surprise, and contempt) on a set of 28 faces presented on slides. The three groups were equally able to distinguish a face displaying affect from one not displaying affect. Naming of emotion was worse in patients with FTD than in patients with AD (correct answers 46% vs. 62%; p = 0.0006) who did not differ significantly from controls (72%). Anger, sadness, and disgust were less recognized in FTD than in AD patients who did not differ from controls, whereas fear and contempt were poorly recognized in both groups of patients compared with controls. These findings argue for different neural substrates underlying the recognition of various basic emotions. Behavioral disorders in FTD may be partly due to an impaired interpretation of the emotional environment. PMID- 10372954 TI - Neuroendocrine responses to intravenous infusion of physostigmine in patients with Alzheimer disease. AB - We have reported that physostigmine, a reversible cholinesterase inhibitor, enhances verbal memory in patients with Alzheimer disease (AD). To elucidate the mechanism of cognition enhancement, plasma hormones were measured during high dose acute and low-dose chronic steady-state intravenous infusions of physostigmine in nine subjects with AD. High-dose hormone responses were measured during and for 24 h after the infusion of physostigmine 1-1.5 mg over 45-60 min. Chronic responses were measured during continuous intravenous infusions of physostigmine at doses (0.5-25 mg/day) that escalated over 2 weeks, and then during 1 week infusion of the dose that optimized cognition (2-12 mg/day) or placebo administered in a randomized, double-blind, cross-over design. A replicable improvement in verbal memory was found in five subjects. High-dose physostigmine infusion that produced noxious side effects resulted in significant elevation above baseline in plasma levels of adrenocorticotrophic hormone (ACTH) (p = 0.0001), cortisol (p = 0.0001), and beta-endorphin (p = 0.0001). Chronic physostigmine administration, in the absence of adverse effects, produced no significant elevation in ACTH (p = 0.08), cortisol (p = 0.70), or beta-endorphin (p = 0.82). These results indicate that high-dose physostigmine activates the hypothalamic-pituitary-adrenal (HPA) axis, likely representing a "stress response." In contrast, cognition-enhancing doses do not produce a peripheral corticosteroid response. Thus, physostigmine-induced memory improvement is independent of the activation of the HPA axis. PMID- 10372955 TI - Outdoor wandering parks for persons with dementia: a survey of characteristics and use. AB - This study aimed to characterize the features of outdoor areas for persons with dementia, and to clarify the relationship between design features, use, and satisfaction with these areas. A national survey of long-term care facilities with outdoor areas investigated the characteristics and features of these areas, and how those related to their perceived impact on their users. Most respondents rated outdoor spaces as very useful, and as having a great benefit for users. The perceived benefit was related to the presence of more design features, such as the presence of gazebos and to the number of activities offered in the area. Despite these positive findings, respondents stated the areas were not used as much as possible and indicated several problems, mostly related to the safety of the residents. The results of this survey can assist facilities in better designing or improving their outdoor areas to increase use and satisfaction. PMID- 10372956 TI - Cataracts and Alzheimer disease. PMID- 10372957 TI - Development of novel monoclonal antibodies for the analysis of functional sites in FGF-2. AB - Fibroblast growth factor 2 (FGF-2) can function as a potent mitogen, as well as a survival factor for a variety of mammalian cell types. The biological effects of FGF-2 are mediated by its interaction with two types of cellular binding sites: (1) high affinity tyrosine kinase receptors; and (2) low affinity heparan sulfate proteoglycans (HSPGs) on the cell surface. Although numerous FGF-2 antibodies have been used previously to analyze its biological actions, few studies have utilized antibodies to analyze domains within FGF-2 involved in its interactions with the two binding sites. In this report, we describe the generation and use of two monoclonal antibodies against human recombinant FGF-2 (254F1 and 256A12) that inhibit FGF-2 function. However, these antibodies appear to target preferentially different domains within the FGF-2 molecule, and therefore differentially influence the interactions of FGF-2 with its low and high affinity receptors. 254F1 is a more effective inhibitor of the high affinity, receptor tyrosine kinase binding site, whereas 256A12 appears to be a better inhibitor of the low affinity, HSPG interactions. We also demonstrate that the two antibodies are potent inhibitors of FGF-2 stimulated vascular cell proliferation, and as such have potential use in the treatment of vascular hyperproliferative diseases. PMID- 10372958 TI - Immunodetection of Xenopus bone morphogenetic protein-1 in adult and embryonic cells. AB - In order to analyze biochemical properties of Xenopus bone morphogenetic protein 1 (XBMP-1), rabbit antiserum (alpha-B1) was raised against a synthetic peptide (P1) corresponding to a hydrophilic N-terminal region. XBMP-1B (Xtld) synthesized in the reticulocyte lysate was successfully immunoprecipitated by this antiserum. This precipitation was completely blocked when P1 was added to the reaction, indicating that alpha-B1 recognized XBMP-1B specifically. In Western blot analysis, two distinct sizes of protein (107 and 34 kD) were detected in hind limbs in metamorphosing animals. Both proteins were detected in various adult tissues such as lung, liver, kidney, heart, muscle, intestine, brain, and testis. The mixing of the liver and muscle extracts, and the following detection of immunoreactive proteins suggested that the 34 kD band was a proteolytic product of the 107 kD protein. In the embryonic extracts from the unfertilized egg (stage 0) to swimming tadpoles (stage 40), a 63 kD protein was detected in addition to the 107 kD protein. We also showed that the 107 kD protein was much more expressed in the animal half of the unfertilized eggs than in the vegetal half, but that it was ubiquitously expressed in the gastrula embryos. We suggest that the 63 and 107 kD proteins correspond to full-length proteins encoded by XBMP-1A and XBMP-1B genes, and these proteins are expressed in embryo and in various adult tissues. PMID- 10372959 TI - Role of TGF-beta1 in platelet-mediated cardioprotection during ischemia reperfusion in isolated rat hearts. AB - Platelets protect myocardium against ischemia-reperfusion injury. This study examined the role of platelet-derived TGF-beta1 in cardioprotection during ischemia-reperfusion. Isolated Sprague Dawley rat hearts were perfused with K-H buffer and subjected to 25 min of global ischemia followed by 30 min of reperfusion. Ischemia-reperfusion resulted in myocardial dysfunction indicated by increase in CPP and LVEDP, and decrease in dLVP. Perfusion of hearts with washed platelets or supernatant of aggregated platelets attenuated (P < 0.01) of myocardial dysfunction following ischemia-reperfusion. Ischemia-reperfusion resulted in a decrease in myocardial TGF-beta1 determined by immunohistochemistry. ELISA showed an increase in latent TGF-beta1, but a decrease in active TGF-beta1. Perfusion of hearts with platelets or aggregated platelet supernatant preserved myocardial TGF-beta1 content upon ischemia reperfusion. Perfusion of hearts with recombinant TGF-beta1 also resulted in cardioprotection following ischemia-reperfusion qualitatively similar to that observed with platelets or aggregated platelet supernatants. RT-PCR analysis showed an increase in myocardial TGF-beta1 mRNA following ischemia-reperfusion. These observations indicate that platelets protect the myocardium against ischemia-reperfusion-mediated dysfunction at least in part by releasing TGF beta1. Increase in both TGF-beta1 mRNA and latent TGF-beta1 does not indicate a defect in the translation of mRNA. Reduction in myocardial TGF-beta1 following ischemia-reperfusion suggests a defect in the conversion of latent TGF-beta1 to active TGF-beta1. PMID- 10372960 TI - The C-terminal region of fibroblast growth factor-1 is crucial for its biological activity and high level protein expression in mammalian cells. AB - We have studied the role of the carboxyl(C)-terminus of fibroblast growth factor(FGF)-1 using prokaryotic and eukaryotic expression systems. The full length FGF-1 protein and its mutants lacking 6- and 9-amino acids at the C terminus IFGF-1 (Cdel6) and FGF-1 (Cdel9), respectively] could be expressed in E. coli cells at the similar levels. The deletion mutants bound very weakly to FGF receptor and to heparin, and did not stimulate DNA synthesis in BALB/c3T3 cells. In contrast to E. coli cells, in NIH3T3 transfectants and L6 transfectants, the protein expression level of FGF-1 (Cdel6) was significantly lower than that of FGF-1, and longer C-terminal deletions further decreased the protein expression levels. However, the level of transcripts in the transfectants and the level of translates in in vitro system were equivalent for all the FGF-1 constructs. Treatment with proteasome inhibitors of the NIH3T3 transfectants expressing FGF 1(Cdel6) increased the protein level six-fold. The results indicate that the C terminus of FGF-I is crucial for its biological activity and high-level expression in mammalian cells and suggest that deletion of the C-terminus of FGF 1 induces its post-translational degradation by proteasome system. PMID- 10372961 TI - Evidence for distinct signaling properties and biological responses induced by the PDGF receptor alpha and beta subtypes. AB - Platelet-derived growth factor (PDGF) acts as a potent mitogen, chemoattractant and survival factor for mesenchymal cells. In addition to its importance in mammalian development, PDGF plays a critical role in physiological repair mechanisms and in the pathogenesis of various proliferative diseases. The biological effects of PDGF are initiated via two related receptor tyrosine kinases, termed alpha and betaPDGF receptors. Recent observations provide increasing evidence for distinct roles of the two PDGF receptor subtypes in both embryogenesis and disease formation. Moreover, characterization of the signal relay mechanisms indicates, that the alpha and betaPDGF receptors are not identical in their ability to bind intracellular effector molecules. Furthermore, the two PDGF receptors initiate overlapping, yet distinct signal transduction pathways. These differences may account for some of the variabilities in biological responses resulting from activation of these two receptors. PMID- 10372962 TI - Cell-density (cycle) dependent silencer of the rat insulin-like growth factor binding protein-2 (IGFBP-2) gene. AB - An element which has a negative effect on transcription has been identified in the 5'-flanking region of the rat insulin-like growth factor binding protein-2 (IGFBP-2) gene. This element was confirmed to be a silencer by truncation or by linking it to a heterologous promoter. The silencer activity disappeared in the growth-arrested BRL-3A cells at the G1/S border by high cell density where the IGFBP-2 production is highly elevated. This observation may represent a novel mechanism through which gene expression is controlled by modulation of a silencer. Taken together, these results suggest a regulatory link between cell growth and IGFBP-2 expression regulated by its silencer. PMID- 10372963 TI - A murine stromal cell line promotes the expansion of CD34high+-primitive progenitor cells isolated from human umbilical cord blood in combination with human cytokines. AB - The in vitro expansion of CD34+ cells is important for clinical applications such as transplantation and gene therapy with CD34+ cells isolated from human umbilical cord blood. In the present study, we developed a xenogenic coculture system involving HUCB-CD34+ cells and a murine stromal cell line, HESS-5 cells, in the presence of recombinant human (rh) cytokines. We examined the effects of combinations of cytokines, such as rh-IL-3, rh-SCF, rh-granulocyte colony stimulating factor (G-CSF), rh-granulocyte-macrophage-CSF and h-erythropoietin (EPO), on the expansion of CD34high+ cells and colony-forming progenitor cells (CFCs). The proliferation of CD34high+ cells and CFCs was dramatically promoted on coculture with HESS-5 cells, and the expansion ratio of the CD34high+ cells showed good correlation with that of high-proliferative potential colony-forming cells (HPP-CFCs). The most potent combination of cytokines in this xenogenic coculture system for the expansion of CD34high+ cells and HPP-CFCs was rh-IL-3 and rh-SCF. The proliferation of CD34high+ cells was supported in the presence of HESS-5 cells with direct cell contact, but not observed in the indirect coculture involving a microporous membrane. Furthermore, we developed a unique coculture method, designated as the bilayer coculture method, involving CD34+ cells and HESS-5 cells using a microporous membrane. This expansion system will be applicable to the expansion of the primitive progenitor cells of HUCB-CD34+ cells and is worthy of consideration for the clinical application of HUCB-CD34+ cells. PMID- 10372964 TI - Receptor binding and biological activity of mammalian expressed sensory and motor neuron-derived factor (SMDF). AB - Sensory and motor neuron-derived factor (SMDF) is a member of the neuregulin family of proteins. SMDF is structurally characterized by a novel N-terminal domain. Using the signal sequence and N-terminal 28 amino acids (the "epitope") of herpes simplex virus type 1 glycoprotein D (gD), we have expressed SMDF as an epitope-tagged protein (gD-SMDF) in 293 cells, and purified it to > 98% homogeneity on a monoclonal anti-gD column. gD-SMDF stimulates human Schwann cell growth and 3H-thymidine incorporation in MCF-7 and T47D human breast tumor cells in vitro. The biological activity of gD-SMDF is consistent with its ability to compete with 125I-labeled heregulinbeta1 peptide (rHRGbeta1(177-244)) to bind to soluble dimeric ErbB receptor-IgG fusion proteins. gD-SMDF binds with low affinity to homodimeric ErbB3-IgG and ErbB4-IgG but with higher affinity to heterodimeric ErbB2/ErbB3-IgG and ErbB2/ErbB4-IgG. Using a SMDF-IgG(Fc) fusion protein we generated a monoclonal antibody (3G11) which binds SMDF, crossreacts with rHRGbeta1(177-244), and neutralizes the in vitro activities of gD-SMDF and rHRGbeta1(177-244) in human Schwann cells. PMID- 10372965 TI - Acidic glycosphingolipids of cock testis elucidated by mass spectrometry and NMR spectroscopy. AB - The main acidic glycosphingolipids (GSLs) of cock testis were identified as GalCer I3-sulfate and gangliosides GM4, GM3, GD3 and GT3. They contained N acetylneuraminic acid as the major sialic acid, and ceramides composed mainly of sphingosine (dl8:1) and C18-24 non-hydroxy fatty acids. Appreciable amounts of hydroxy fatty acids were detected only in the GM4 preparation. PMID- 10372967 TI - In-situ hybridization localized MUC7 mucin gene expression to the mucous acinar cells of human and MUC7-transgenic mouse salivary glands. AB - MUC7 gene that encodes the low molecular weight human salivary mucin MG2, was previously shown to be expressed in tissue-specific manner in normal human salivary glands and in salivary glands of transgenic mice carrying the MUC7 transgene. The purpose of this study was to examine the cell specificity of MUC7 expression in human and transgenic mice salivary glands. To localize the MUC7 transcripts, we used in-situ hybridization in combination with Tyramide Signal Amplification procedure. The results clearly showed that in both the human and transgenic mice salivary gland tissue sections, MUC7 transcripts were localized only in mucous acinar cells; no signals were found in serous acinar cells or any other cell types present in these salivary glands. PMID- 10372968 TI - Comparison of lectin-binding patterns between young adults and older rat glycoproteins in the brain. AB - Glycoproteins in the soluble fraction and in the membrane fraction of various portions of brains and spinal cords, obtained from 9-week-old rats and 29-month old rats, were comparatively analyzed by SDS-polyacrylamide gel electrophoresis and lectin staining. The glycoprotein patterns of each brain part showed marked differences by the age of donors. The most prominent evidence in the soluble fractions of white matter, basal ganglia, and spinal cord detected by WGA is that the glycoproteins with an apparent molecular weight of 123K and 115K have increased in the aged rats. In addition, the reactivity of 115K with Con A and PNA has also increased in the aged rats. On the other hand, reactivity of an apparent molecular weight of 115K with WGA has increased in the membrane fractions of white matter, basal ganglia, hippocampus, cerebellum, and spinal cord from the aged rats. In contrast, by MAM, which is specific for Siaalpha2- >3Gal linkage, an apparent molecular weight of 115K has been detected only in the membrane fraction of cerebellum and it has decreased in the aged rats. Reactivity of an apparent molecular weight of 133K and 125K in the membrane fractions of white matter and basal ganglia with LCA has decreased in the aged rats. In contrast, reactivity of the front band with LCA and AAL has increased and that of 130K with AAL has decreased in spinal cord from the aged rats, respectively. These results indicate that the glycosylation state of the protein in the brain changes during aging. PMID- 10372966 TI - A novel second isoenzyme of the human UDP-N-acetylglucosamine:alpha1,3-D mannoside beta1,4-N-acetylglucosaminyltransferase family: cDNA cloning, expression, and chromosomal assignment. AB - We isolated a novel cDNA encoding a second isoenzyme of UDP-N acetylglucosamine:alpha1,3-D-mannoside beta1,4-N-acetylglucosaminyltransferase (GnT-IV; EC 2.4.1.145). The nucleotide and deduced amino acid sequences of the cDNA were homologous to those of the previously cloned human GnT-IV cDNA (63% and 62% identity, respectively). The new cDNA is also confirmed to express GnT-IV activity, suggesting that two isoenzymes of human GnT-IV exist. Although genomic Southern analysis suggested that both genes exist in many mammalian species and the chicken, northern analysis revealed that both genes are expressed in different ways in human tissues. This is the first report concerning the gene family of an N-acetylglucosaminyltransferase in mammals. PMID- 10372969 TI - Identification of antibodies directed against O-acetylated sialic acids in visceral leishmaniasis: its diagnostic and prognostic role. AB - A significantly increased O-acetylated sialic acid (O-AcSA) binding fraction was purified from serum of visceral leishmaniasis (VL) patients by affinity chromatography on immobilized bovine submaxillary mucin (BSM) and found to be immunoglobulin in origin. The serodiagnostic and prognostic potential of BSM as a capture antigen was established by ELISA with no cross reactivity with coendemic diseases like malaria, tuberculosis, leprosy, chagas disease and cutaneous leishmaniasis; however, a strong cross reactivity was present with trypanosomiasis patients. In 56 clinically diagnosed VL patients, the BSM-ELISA was compared with diagnosis by microscopy using Giemsa stained tissue smears and direct ELISA using crude parasite antigen (parasite-ELISA); 49/56(87.5%) and 5/56(9.0%) were positive and negative respectively by all 3 methods. The BSM ELISA failed to diagnose 2/56(3.5%) patients which were biopsy and parasite-ELISA positive. The prognostic potential of the BSM-ELISA in 18 longitudinally monitored VL patients before and after conventional antimonial treatment showed a significant decrease in anti O-AcSA titres in drug responsive patients whereas anti O-AcSA levels persisted in drug unresponsive patients. The IgG subclass distribution of antibodies directed against O-AcSA showed increased IgG2 levels in VL patients as compared to healthy controls. The BSM-based ELISA holds great promise as a serodiagnostic and prognostic assay for VL. PMID- 10372970 TI - Sialyl Lewis(x) expression on IgG in rheumatoid arthritis and other arthritic conditions: a preliminary study. AB - Both infiltrating leukocytes and soluble immunoglobulin form aggregates in synovial fluid during the inflammatory process in rheumatoid arthritis (RA). Some of these changes are probably mediated by the adhesion molecule, E-selectin, which increases its expression with disease activity. As glycosylation changes in IgG in RA are well established, the current study was undertaken to measure the expression of the carbohydrate antigen sialyl Lewis x (sLe(x)), on IgG in RA. sLe(x) is a major ligand for E-selectin. Using a recently developed ELISA, sLe(x) expression was determined in IgG isolated from 8 healthy individuals, 20 RA sufferers (10 early and 10 with more long-standing disease) and 20 patients with other rheumatic conditions (osteoarthritis, ankylosing spondylitis, systemic lupus erythematosus). S Le(x) expression on IgG was elevated above the reference range in all but one of the RA patients and this change was highly significant (P < 0.0006). Expression of this antigen on IgG was also significantly different from normal in the other arthritic groups (P < 0.02), but the changes were much less than that observed for RA. In early RA, sLe(x) was inversely correlated with parameters used to measure disease activity. This was not observed with the established RA, where there was weak positive association. These preliminary results indicate that a change in sLe(x) expression on IgG is an early finding in the development of RA, which may be important in the development of the disease or for predicting its outcome. PMID- 10372971 TI - Altered expression of sialylated carbohydrate antigens in HT29 colonic carcinoma cells. AB - To determine whether cell growth conditions impacted carbohydrate expression, HT29 cells were gradually transferred from a conventional glucose-containing media to a glucose-free galactose containing media. Indirect immunofluorescence on acetone fixed cells showed increased expression of sialyl Lewis A antigen (CA19-9), sialyl Lewis C (DUPAN2) and Tn/sialyl-Tn on the surface of HT29 cells grown in the glucose-free galactose containing media compared to those grown in the glucose containing media. Sialyltransferases responsible for the synthesis for these sialylated epitopes were Increased in the galactose-fed HT29 cells. Media overlying the cells was subjected to isopycnic ultracentrifugation in cesium chloride and the fractions derived from both glucose and galactose media with equivalent buoyant densities of 1.56 g/L, which are predicted to contain mucin glycoforms, were further separated by HPLC using a Mono-Q anion exchange column. The chromatograph of eluent from the sample derived from the cells growing in the galactose containing media showed an increased peak that reacted with the anti-sialyl Lewis A antibody, CA19-9. These results show that alteration of in vitro culture conditions may cause HT29 colonic carcinoma cells to alter the expression of sialylated carbohydrates. PMID- 10372973 TI - Electrophysiological changes in rat ventricular and atrial myocardium at different stages of experimental diabetes. AB - Action potential configuration in ventricular and atrial myocardium, as well as rate-dependent changes in ventricular action potential duration (APD) were studied and compared in healthy and diabetic rats. Diabetes was induced by a single injection of streptozotocin (STZ, 65 mg kg(-1) i.v.). Conventional microelectrode techniques were applied to record action potentials after the establishment of diabetes (2, 6, 10 and 18 weeks after STZ-treatment). Untreated age-matched animals were used as controls. Both depolarization and repolarization were significantly retarded following STZ-treatment. However, the time course of development of diabetic changes in atrial and ventricular myocardium was different. APD was significantly lengthened from week 2 of diabetes in ventricular, but only from week 6 in atrial preparations. In atrial myocardium, lengthening of APD was more pronounced at early rather than late phases of repolarization. The maximum rate of depolarization (Vmax) was significantly reduced from the 6th week of diabetes in both preparations. No differences were observed in action potential amplitude (except at week 18) and in the resting membrane potential in diabetic rats. Diabetic ventricular preparations showed a positive APD-frequency relationship at any level of repolarization, in contrast to control muscles, where APD25 and APD50 values lengthened. But APD75 and APD90 values were not changed significantly with increase in the pacing frequency. The results indicate that development of diabetic alterations are not fully identical in atrial and ventricular myocardium of the rat, probably owing to differences in density and kinetics of ionic currents responsible for atrial and ventricular action potentials. PMID- 10372972 TI - Post-partum prolongation of the atrial repolarization in rabbit. AB - Female sexual steroids are known to modify the expression of various K+ channels and thus they can alter cardiac repolarization. In the present work, using conventional microelectrode techniques, action potential characteristics were studied in atrial myocardium isolated from virgin, late pregnant, early (1-3 days) post-partum and late (2-3 weeks) post-partum rabbits. No changes in action potential configuration were observed during pregnancy. However, the duration, overshoot and amplitude of action potentials were significantly increased in the early (1-3 days) post-partum period. Resting potential and maximum rate of depolarization remained unchanged. The observed changes were transient, normal action potential characteristics were obtained at weeks 2-3 post-partum. 4 aminopyridine (1 mmol L(-1)). caused a marked lengthening of action potential duration in all preparations obtained from non-pregnant and pregnant rabbits, whereas this 4-aminopyridine-induced prolongation was moderate in those preparations excised from the hearts of early post-partum animals. Action potential configuration was not affected by pinacidil (10 micromol L(-1)) or glibenclamide (5 micromol L(-1)) in non-pregnant or pregnant animals. In preparations obtained from early post-partum rabbits, pinacidil significantly shortened action potential duration, which was reverted by glibenclamide. The lengthening of action potential duration together with the decreased sensitivity to 4-aminopyridine observed in early post-partum animals may probably be caused by reduction of the transient outward K+ current at this stage. The results also suggest that electrophysiological alterations in the early post-partum period may probably be more pronounced than those associated with pregnancy itself. PMID- 10372974 TI - Sustained vessel dilation induced by increased pulsatile perfusion of porcine carotid arteries in vitro. AB - Arterial pulse pressure (PP) increases with exertional stress and ageing, and can modify vessel diameter in smaller vessels. To test if PP must exceed a certain range to influence vessel diameter, and determine if such effects are endothelium dependent or intrinsic to vascular viscoelasticity, eight fresh excised porcine carotid artery segments were perfused with modified Krebs-Henseleit by a servo controlled system generating physiological arterial pressure waveforms. In a separate group of vessels (n = 10), the endothelium was mechanically removed. Vessel external diameter was measured by video edge-detection. Vessels partially preconstricted with noradrenaline were perfused at 9 mL min(-1) mean flow, at mean pressure of 90 or 120 mmHg, and zero PP. PP alone was then increased to 40, 70, or 120 mmHg at 1 Hz cycling rate for 5 min, then returned to zero and vessel diameter measured immediately thereafter. The protocol was repeated after 10-20 min stabilization. Mean vessel diameter rose proportionally with PP only once PP exceeded 40 mmHg, with maximal increases of 6-9% at a PP of 120 mmHg. Similar responses were obtained in vessels with and without a functional endothelium, at both mean pressures. Thus, when exposed to higher than normal resting PP, conduit arteries dilate owing to the stress-relaxation response of their viscoelastic wall. This mechanism of PP-mediated vascular dilatation may contribute to enhanced organ perfusion when small resistance arteries are already dilated. PMID- 10372975 TI - Isotonic and hypertonic sodium loading in supine humans. AB - The hypothesis that hypertonic saline infusion induces a greater natriuresis than infusion of the same amount of sodium as isotonic saline was tested in 8 supine subjects on fixed sodium intake of 150 mmol NaCl day(-1). Sodium loads equivalent to the amount of sodium contained in 10% of measured extracellular volume were administered intravenously over 90 min either as isotonic saline or as hypertonic saline (850 mmol L(-1)). A third series without saline infusion served as time control. Experiments lasted 8 h. Water balance and sodium loads were maintained by replacing the excreted amounts every hour. Plasma sodium concentrations only increased following hypertonic saline infusion (by 2.7 +/- 0.3 mmol L(-1)). Oncotic pressure decreased significantly more with isotonic saline (4.1 +/- 0.3 mmHg) than with hypertonic saline (3.2 +/- 0.2 mmHg), indicating that isotonic saline induced a stronger volumetric stimulus. Renal sodium excretion increased more than a factor of four with isotonic and hypertonic saline but also increased during time control (factor of three). Cumulated sodium excretions following isotonic (131 +/- 13 mmol) and hypertonic saline (123 +/- 10 mmol) were statistically identical exceeding that of time control (81 +/- 9 mmol). Plasma angiotensin II decreased in all series but plasma ANP concentrations and urinary excretion rates of endothelin-1 remained unchanged. In conclusion, hypertonic saline did not produce excess natriuresis. However, as the two loading procedures induced similar natriureses during different volumetric stimuli, part of the natriuresis elicited by hypertonic saline could be mediated by stimulation of osmoreceptors involved in renal sodium excretion. The supine position does not provide stable time control conditions with regard to renal excretory function. PMID- 10372976 TI - Thermogenic response to adrenaline during restricted blood flow in the forearm. AB - To elucidate the underlying mechanism behind the thermogenic effect of adrenaline in human skeletal muscle, nine healthy subjects were studied during intravenous infusion of adrenaline. Restriction of blood flow to one forearm was obtained by external compression of the brachial artery, to separate a direct metabolic effect of adrenaline from an effect dependent on increased blood flow. The other arm served as the control arm. In the control arm, the forearm blood flow increased 4.7-fold (from 2.0 +/- 0.3 to 9.3 +/- 1.5 mL 100 g(-1) min(-1), P < 0.001) during the adrenaline infusion. Adrenaline significantly increased forearm oxygen consumption (from 4.7 +/- 2.1 to 7.0 +/- 3.6 micromol 100 g(-1) min(-1), P < 0.025). In the arm with restricted blood flow, the forearm blood flow increased 2.9-fold (from 1.6 +/- 0.3 to 4.6 +/- 0.8 mL 100 g(-1) min(-1), P < 0.002) but the forearm oxygen consumption did not increase (baseline period: 5.6 +/- 2.3 micromol 100 g(-1) min(-1), adrenaline period: 6.1 +/- 2.1 micromol 100 g(-1) min(-1), P = 0.54). The experimental design and the difficulties in interpretation of the result are discussed. The results give evidence for the hypothesis that the vascular system plays a key role in the thermogenic effect of adrenaline in skeletal muscle in vivo. PMID- 10372978 TI - Effects of age, sex and physical exercise on the phagocytic process of murine peritoneal macrophages. AB - Different stages of the phagocytic process, i.e. chemotaxis, ingestion of latex beads and superoxide anion production, were measured in peritoneal macrophages from young (12 +/- 2 weeks old) and old (60 +/- 2 weeks old) male and female BALB/C mice, which were subjected to an acute bout of exercise (swimming until exhaustion) or to a period of training exercise (90 min of swimming each day during 20 days). Sedentary male and female mice as well as young and old animals were used as sex and age controls. The results show that both acute and training exercise stimulate the phagocytic process of murine peritoneal macrophages. The macrophage functions from sedentary controls were lower in the old than in the young animals. The chemotaxis and ingestion capacities of macrophages were higher in the female young and old sedentary mice than in their male counterparts. After exercise, the stimulation of the phagocytic process was higher in old and female mice than in young and male animals. In addition, serum corticosterone levels were measured in order to investigate the relations between stress and macrophage activity. Old and female mice as well as animals subjected to exercise showed, in general, higher corticosterone levels than young, male and sedentary animals. PMID- 10372977 TI - Modulation of Na+ x H+ exchange by hydrostatic pressure in isolated bovine articular chondrocytes. AB - The effects of increased hydrostatic pressure on Na+ x H+ exchange activity in bovine articular chondrocytes have been characterized. Chondrocytes were isolated from the cartilage matrix and the cells were loaded with the pH-sensitive fluorophore BCECF. Cells were acidified by ammonium rebound and the rate of recovery of pHi back to control levels was determined using cuvette fluorimetry. The application of hydrostatic pressure (1-300 atm) to cells within the fluorimeter was found to stimulate the rate of recovery from acidification, recorded as proton fluxes, in MOPS buffered media. This increase was dependent on the presence of extracellular Na+ ions and was inhibited by the Na+ x H+ exchange inhibitor EIPA. The pressure-stimulated increase in H+ flux is therefore mediated completely by Na+ x H+ exchange. In addition, the stimulation could be abolished by the kinase inhibitor staurosporine, was not additive with the stimulation of Na+ x H+ exchange elicited by the addition of serum and was unaffected by low concentrations of the myosin light chain kinase inhibitor ML-7. We therefore conclude that hydrostatic pressure activates Na+ x H+ exchange in this cell type by a pathway which involves direct phosphorylation of the transporter protein itself. This is the first demonstration of the activation of Na+ x H+ exchange by hydrostatic pressure and the relevance of this finding to the biology of cartilage tissue is discussed. PMID- 10372980 TI - The mechanotransduction of the crayfish stretch receptor neurone can be differentially activated or inactivated by local anaesthetics. AB - The effect of the local anaesthetics lidocaine, its meta-isomer, LL33, bupivacaine, tetracaine and procaine on the transducer properties of the stretch receptor neurone of the crayfish Pacifastacus leniusculus was investigated using a two microelectrode voltage clamp. Lidocaine increased the receptor current whereas LL33, bupivacaine and tetracaine reduced the receptor current in a reversible dose-dependent way. Procaine did not affect the receptor responses. The onset of the effect was generally slow in the order of minutes. Lidocaine increased the conductance of the mechanotransducer 50 +/- 7% (mean +/- SD, n = 4) and changed the reversal potential -8 +/- 1 mV (mean +/- SEM, n = 8), which indicates a major K+ conductance increase through the mechanosensitive channels. The other local anaesthetics increase the K+ conductance of the mechanotransducer without increasing the total conductance, which suggests that only P(Na)/P(K) is changed. These substances seem to have a Ca2+ dependent effect on the gating properties of the mechanosensitive channels in addition to their effect on the permeability through the channels as compared with lidocaine. All local anaesthetics investigated decreased the leak conductance of the receptor neurone. The effects of local anaesthetics on the mechanosensitive channels whether activating or blocking is correlated to the oil:water distribution coefficients and their relative hydrophobicity/hydrophilicity ratio. The results are consistent with the hypothesis that the local anaesthetic effect is mediated by changes in the lipid phase of the membrane. PMID- 10372981 TI - The tachykinins neurokinin A and substance P, but not neurokinin B, stimulate contraction of isolated muscle cells from rat small intestine. PMID- 10372979 TI - The effects on net fluid transport of noxious stimulation of jejunal mucosa in anaesthetized rats. AB - A major aim of the present study was to investigate whether exposing the jejunal mucosa to a noxious stimulus induces a net fluid secretion by activating the enteric nervous system (ENS) and, if so, to what extent an axon reflex was involved. Net fluid transport was measured in vivo with a gravimetric method. The intestinal mucosa was exposed to an isotonic solution with an unphysiologically low pH (1.0). This evoked a fluid secretion, which was markedly attenuated by giving hexamethonium (nicotinic receptor antagonist) i.v. or exposing the intestinal serosa to lidocaine (local anaesthetic). Atropine (muscarinic receptor antagonist) had no effect. Luminal acid evoked a fluid secretion of the same magnitude in acutely denervated segments and in segments denervated about 3 weeks prior to the experiments. Luminal capsaicin (1.6-16 mM) did not influence jejunal net fluid transport. A second aim of the study is to investigate the effect of nifedipine (Ca channel blocker of L-type) on the acid-induced fluid secretion. Nifedipine markedly attenuated acid-induced fluid secretion. In contrast to cholera toxin-evoked secretion, the nifedipine effect was not mediated via 5 hydroxytryptamine (5-HT) as judged by measurements of 5-HT release into the intestinal lumen and the lack of effect of granisetron (5-HT3 receptor antagonist). It is concluded that the net fluid secretion evoked by hydrochloric acid in the small intestine is mainly mediated via an intramural reflex in the ENS. No experimental evidence was obtained for the involvement of an axon reflex. The site of action of the calcium channel blocker is tentatively discussed. PMID- 10372982 TI - Brain cooling in diving seals. PMID- 10372983 TI - Effects of irrigation fluid temperature on choroidal circulation during vitrectomy. AB - PURPOSE: To investigate the effects of irrigation temperature on choroidal circulation during vitrectomy. METHODS: After anesthetized albino rabbits underwent closed vitrectomy, choroidal blood flow was monitored while the vitreous cavity was irrigated with solutions (BSS plus) of various temperatures (6-42 degrees C). Irrigation pressure was maintained at 15 mmHg. A non-contact Doppler laser flowmeter was used to measure choroidal blood flow. Temperature change at the retina and mid-vitreous was also measured during irrigation. RESULTS: Choroidal blood flow showed a downward peak when perfused at 34 degrees C. As irrigation temperatures deviated above or below 34 degrees C, the blood flow increased. However, when irrigation temperature was below 16 degrees C, the blood flow decreased as the temperature declined. The temperature at the retina was maintained at a relatively constant level when irrigation was between 30 degrees C and 40 degrees C. CONCLUSION: The choroid acts as a thermostat to minimize intraocular temperature fluctuations by changing its blood flow. PMID- 10372984 TI - Stimulation of active sodium-potassium transport by hydrogen peroxide in cultured rabbit nonpigmented ciliary epithelium. AB - PURPOSE: Studies were conducted to examine the effect of hydrogen peroxide on active sodium-potassium transport in a cell line derived from nonpigmented ciliary epithelium of the rabbit eye. METHODS: Studies were carried out using a rabbit nonpigmented ciliary epithelium cell line. 86Rb uptake by intact cells was measured in the presence or absence of ouabain. The ouabain-sensitive potassium (86Rb) uptake rate was used as an index of the rate of active sodium-potassium transport. Cell sodium content was measured by atomic absorption spectrophotometry. Na,K-ATPase activity was determined by measuring ATP hydrolysis in the presence or absence of ouabain, using membrane material isolated by centrifugation of cell homogenates. RESULTS: Ouabain-sensitive potassium (86Rb) uptake rate measured in cells that had been preincubated with 200microM hydrogen peroxide for either 30 min or 60 min was increased to 196% and 181% of the control uptake rate, respectively. Lesser concentrations of hydrogen peroxide caused lesser degrees of stimulation. 200microM hydrogen peroxide caused an increase of cell sodium content. Such a change of cell sodium content is likely to be responsible, at least in part, for the observed stimulation of active sodium-potassium transport. However, the response may also be partly dependent on activation of a protein kinase since the serine/threonine protein kinase inhibitors staurosporine (1microM) and H-89 (20microM) were both found to prevent the stimulatory effect of 200microM hydrogen peroxide on ouabain sensitive potassium (86Rb) uptake. Interestingly, neither H-89 nor staurosporine prevented the elevation of sodium content in cells that received 200microM hydrogen peroxide. CONCLUSIONS: Taken together, these findings suggest a low concentration of hydrogen peroxide causes increased sodium entry into the cell and also activates a protein kinase-dependent mechanism for sodium pump stimulation. The protein kinase-dependent mechanism does not appear to be triggered by an increased rate of sodium entry since staurosporine did not prevent the stimulation of ouabain-sensitive potassium (86Rb) uptake elicited by an increase in sodium permeability caused by amphotericin B. PMID- 10372986 TI - Conformational study of N(epsilon)-(carboxymethyl)lysine adducts of recombinant alpha-crystallins. AB - PURPOSE: Lens proteins underwent nonenzymatic glycation, and the advanced glycation end products (AGEs) were detected by immunological assays. One of the major AGE structures is N(epsilon)-(carboxymethyl)lysine (CML). Since the involvement of AGEs in the pathogenesis of diabetic complications is speculated, the effects of CML formation on proteins were studied. METHODS: CML adducts were generated in recombinant alphaA- and alphaB-crystallins by incubation with glyoxylic acid and NaBH3CN. SDS-PAGE and size exclusion chromatography were used to detect subunit degradation and high-molecular-weight (HMW) aggregation. Conformational change was determined by fluorescence and circular dichroism (CD) measurements. The chaperone function was studied by DTT-induced aggregation of insulin. RESULTS: Lysine modification was estimated to be 60-90% depending on the conditions of incubation. No subunit degradation or HMW aggregation was observed. Fluorescence and CD measurements detected a conformational change in CML adducts. Measurements of chaperone-like activity, however, indicated that the formation of CML increased the protein's ability to protect insulin against DTT-induced aggregation. CONCLUSIONS: Although CML adducts of alphaA- and alphaB-crystallins, the major AGE structures formed in vitro, changed protein conformation, no subunit degradation and HMW aggregation were observed. Moreover, the CML adducts increased chaperone-like activity of both alphaA- and alphaB-crystallins. The results suggest that CML formation alone may not play a major role in protein aggregation and lens opacity. PMID- 10372985 TI - Phosphorothioate oligonucleotides induction into experimental choroidal neovascularization by HVJ-liposome system. AB - PURPOSE: The purpose of this study was to determine whether the inactivated hemagglutinating virus of Japan (HVJ)-liposome method can induce phosphorothioate oligonucleotides effectively into an experimentally-induced choroidal neovascularization of rats. We also examined whether antisense phosphorothioate oligonucleotides against VEGF could be induced into choroidal neovascularization as a therapeutic agent by the HVJ-liposome method. METHODS: The experiments were conducted on a rat model of choroidal neovascularization. FITC-labeled phosphorothioate oligonucleotides were coencapsulated in liposomes. The liposomes were coated with the envelope of inactivated HVJ and injected into the vitreous cavity following photocoagulation of pigmented rat eyes. The eyes were removed following injection, fixed, frozen and cut into thin sections. Induction of oligonucleotides was observed under a laser confocal scanning microscope for fluorescence and the development of choroidal neovascularization was evaluated histopathologically. RESULTS: Phosphorothioate oligonucleotides were effectively induced into ganglion cells and into the cells of the choroidal neovascularization induced by laser photocoagulation. Highly effective induction of oligos was observed 3 to 14 days after intravitreal injection of HVJ-liposomes after which the level decreased. Antisense oligonucleotides against VEGF were induced specifically into cells in the choroidal neovascularization, however neovascularization was still observed. CONCLUSIONS: Phosphorothioate oligonucleotides can be effectively induced into ganglion cells, and specifically into cells in choroidal neovascularization. Although antisense oligonucleotides against VEGF failed to prevent choroidal neovascularization, the HVJ-liposome method provided a highly effective means of inducing antisense oligos for in vivo antisense therapy. PMID- 10372987 TI - 3-Fluoro-3-deoxy-D-galactose: a new probe for studies on sugar cataract. AB - PURPOSE: Aldose reductase (AR) activity and flux through the polyol pathway can conveniently be monitored in dog lenses by measuring the metabolism of 3-fluoro-3 deoxy-D-glucose by 19F nuclear magnetic resonance (NMR) spectroscopy. Since AR has broad substrate specificity and preferentially utilizes galactose over glucose as substrate, the ability of AR to utilize 3-fluoro-3-deoxy-D-galactose (3-FDGal) as substrate as well as the metabolism of 3-FDGal in intact dog lens and cultured lens epithelial cells has been investigated. METHODS: The suitableness of 3FDGal as a substrate was examined by incubating 3FDGal with purified dog lens aldose reductase in the presence of an NADPH generating system or with galactitol dehydrogenase in the presence of NAD+. Dog lenses and dog lens epithelial cells were cultured in 3-FDGal medium with and without the AR inhibitor AL 1576. Metabolism was studied using 19F NMR. RESULTS: AR activity with 3-FDGal as substrate is higher than that with D-galactose and its Km of 4.2 mM is ca 10-fold higher than that of D-galactose. Purified dog lens AR incubated with 3-FDGal resulted in the formation of 3-fluoro-3-deoxy-D-galactitol. Galactitol formation was prevented by the addition of AL 1576. Incubation of 3 FDGal with galactitol dehydrogenase resulted in the formation of 3-fluoro-3-deoxy D-galactonic acid. Dog lenses cultured in 3-FDGal medium formed NMR peaks corresponding to 3-fluoro-3-deoxy-D-galactitol and 3-fluoro-3-deoxy-D-galactonic acid. The presence of AL 1576 inhibited the formation of galactitol but not galactonic acid. Lens epithelial cells cultured in 3-FDGal medium formed only 3 fluoro-3-deoxy-D-galactitol. These cells developed multiple cytoplasmic vacuoles which was prevented by the aldose reductase inhibitor AL 1576. CONCLUSIONS: The high affinity of this fluorinated sugar for aldose reductase makes this an excellent probe for investigating aldose reductase activity in dog lens tissues. PMID- 10372988 TI - Elements regulating the transcription of human interstitial retinoid-binding protein (IRBP) gene in cultured retinoblastoma cells. AB - PURPOSE: To identify cis-acting elements and trans-acting factors involved in the expression of human IRBP gene. METHODS: Transient transfection of WERI-Rb1 and HeLa cells, DNase 1 footprinting, gel mobility-shift assay and yeast one-hybrid system were used to study the regulatory elements that are involved in the expression of human IRBP gene. RESULTS: A region between -1620 and -1411 was shown to have enhancer properties. Using nuclear extracts from WERI-Rb1 and HeLa cells, four footprints were identified in the proximal promoter region (-206 to +68). The core promoter element IP1 binds to OTX2 in the yeast one-hybrid system. By cotransfecting HeLa cells, OTX2 could transactivate the irbp promoter. The functions of IP2 (from -119 to -86) and IP3 (from -183 to -147) remain to be determined. The region containing the HeLa cell-specific footprint IP4 (from -202 to -180) could silence the OTX2 transactivation of the irbp promoter. CONCLUSION: The 5'-flanking region of irbp contains an enhancer sequence. The possible silencer upstream from the core promoter may serve to suppress expression of irbp in HeLa cells. When the proximal promoter is used to identify binding proteins in a human retina library by the yeast one hybrid system, nine of the identified clones contained the cDNA sequence for the homeodomain protein OTX2. Since no clones for the homeodomain protein CRX were found, and since OTX2 can transcriptionally activate irbp in normally non-expressing HeLa cells, it is possible that OTX2 rather than CRX is the transcriptional activator for irbp in human photoreceptors. PMID- 10372990 TI - Immunohistochemical identification of androgen receptors in human lacrimal glands. AB - PURPOSE: Androgens are thought to play a role in the regulation of the human lacrimal gland. Androgen receptor mRNA has been isolated from human lacrimal tissue; however, it is not known which cell(s) in human lacrimal tissue may contain androgen receptors. This study is an immunohistochemical investigation of the location and distribution of androgen receptors in human lacrimal tissue. METHODS: Formalin-fixed, paraffin-embedded human lacrimal gland tissues were subjected to established antigen retrieval techniques. This was followed by routine immunohistochemical staining, employing one of two anti-human androgen receptor monoclonal antibodies, each specific for a different antigenic epitope within the receptor molecule. RESULTS: The two anti-human androgen receptor monoclonal antibodies demonstrated similar staining patterns in adjacent tissue sections from the same human lacrimal gland specimens. Specific staining for androgen receptors was observed in the nucleus and cytoplasm of lacrimal acinar cells, as well as in lacrimal duct cells. Both the intensity of staining and the number of cells demonstrating staining varied among specimens. We also observed staining for androgen receptors in interstitial and inflammatory cells distributed between lacrimal acinar units in some specimens. CONCLUSIONS: Androgen receptors are located in human lacrimal gland acinar cell nuclei as observed in other animals. However, the detection of androgen receptors in lacrimal interacinar interstitial and inflammatory cells suggests that androgens may play a role in modulating the activities of cells other than lacrimal cells within the human lacrimal gland. PMID- 10372991 TI - Cloning of a major human retinal pigment epithelial lysosomal aspartic protease and mapping its transcriptional start sites. AB - PURPOSE: It has been proposed that the major proteolytic enzyme responsible for the proteolysis of photoreceptor outer segments is an aspartic protease similar or identical to cathepsin D. The aim of this study was to determine if the major retinal pigment epithelial (RPE) aspartic protease was cathepsin D, to study its distribution and investigate its transcription start sites (TSS). METHODS: An RPE cDNA library was screened at low stringency for the presence of aspartic proteases. DNA sequencing was used to analyze the identified clones. The expression of cathepsin D mRNA was analyzed by Northern blot while immunohistochemistry was used to demonstrate cathepsin D related immunoreactivity in an eye section. RNase protection assay was used to map the Cathepsin D TSS in RPE cells. RESULTS: The aspartic protease identified in the RPE cDNA library was identical to the DNA sequence of cathepsin D found in other tissues. Northern blot analysis of a wide range of cell types showed that the highest level of cathepsin D mRNA expression was found in RPE cells which was similar only to cathepsin D expression in MCF7 breast cancer cells. Immunohistochemical staining of a human retina confirmed the inherent nature of high cathepsin D expression in RPE cells. An RNase protection assay demonstrated two major cathepsin D TSS in RPE cells. One of them was identical to the TSS responsible for the constitutive expression of cathepsin D and the other, a TATA box-controlled TSS, was identical to a TSS found in MCF7 cells. Cathepsin D expression was not enhanced by the phagocytosis and digestion of rod outer segments (ROS) and ROS phagocytosis did not induce the use of other cathepsin D TSS in the RPE cells. CONCLUSION: The major aspartic protease identified in RPE cells was cathepsin D. In addition, it was demonstrated that the high level of cathepsin D expression in RPE cells is the intrinsic nature of these cells and that it is linked to a TATA box controlled TSS. This TSS has previously been described as an estrogen regulated TSS and further studies are required to identify the RPE-specific inducer or indirect factors that may be responsible for the enhanced expression of cathepsin D in the RPE cells. PMID- 10372989 TI - Correlation between prevention of cataract development by disulfiram and fates of selenium in selenite-treated rats. AB - PURPOSE: We found a new pharmacological effect of disulfiram (DSF) against rat pups with cataract induced by selenite injection. The possible reactive mechanism is discussed in this present paper. METHODS: Wistar male and female rats aged 13 and 30 days, and male rats aged 7 weeks were used this present study. Cataract was induced by injection of selenite (19 micromol/kg, s.c.) to 13-day old rats. The lens opacification was monitored by using the slit lamp equipped with an anterior eye segment analysis system (EAS-1000, Nidek). The selenium contents in rat organs were detected fluorimetrically. Liposomes containing DSF (DSF liposomes) were prepared by the reverse-phase evaporation method. Rat pups were instilled 5 microl of DSF-liposomes into both eyes 4 times per day. Adult rats were administered with DSF suspensions (100 mg/kg) by nasal cannulation. The changes of plasma concentration of diethyldithiocarbamate (DDC), which was a metabolite of disulfiram, were determined by HPLC method. RESULTS: Intraocular treatment with DSF-liposomes prevented the onset of cataract development in rat pups injected with sodium selenite. Treatment with DSF also significantly reduced the selenium contents in plasma at 1 h post-treatment and in the eye at 96 h post treatment. No significant differences of selenium content in liver and kidney were observed in 13-day old rats instilled with DSF-liposomes or DSF free liposomes. Cataract could not be induced in the 30-days-old rats with the same dose of selenite (19 micromol/kg) and the liver, kidney, and especially eye of the older rats had lower levels of selenium than 13-day old rats. Diethyldithiocarbamate (DDC), an active metabolite of DSF, was decreased in the plasma following a subcutaneous injection of sodium selenite. The selenium concentration in the plasma was decreased by the intranasal administration of DSF suspensions. CONCLUSIONS: Instillation of DSF-liposomes into the eyes of rat pups given a subcutaneous injection of sodium selenite to induce cataracts prevented the formation of cataracts. The mechanism of inhibition may have resulted from a decreased level of selenium in the eyes following the treatment with DSF. On the other hand, as it is known that the cataracts may result from selenite-induced oxidative stress in the lens, DSF and DDC may react as anti-oxidants. PMID- 10372992 TI - Mutant herpes simplex virus-mediated suppression of retinoblastoma. AB - PURPOSE: To test the ability of a mutant herpes simplex virus (HSV) hrR3 to inhibit growth of Y79 human retinoblastoma in vitro and in vivo. METHODS: Cultured Y79 cells were infected with multiplicities of infection (MOI) ranging from 0.004 to 0.1 of hrR3. Surviving cells were counted using trypan blue dye exclusion. Using X-gal staining, expression of the lacZ gene was examined in vitro on day 3 postinfection to evaluate viral replication. Nude mice harboring Y79 tumors subcutaneously received an intraneoplasmic injection of 5 x 10(7) plaque-forming units of hrR3. The tumor sizes were measured weekly. Expression of the lacZ gene was also examined on one week postinfection. RESULTS: There are 31% and 13% cells surviving in cultured Y79 cells infected by hrR3 at an MOI of 0.1 on days 3 and 5 postinfection respectively compared to those of mock-infected cells. Also more than 70% of Y79 cells were stained with X-gal at an MOI of 0.1 which demonstrated active viral replication in vitro. Virus-treated subcutaneous tumors were smaller than control tumors (p<<0.05, Student's t-test) on days 14, 21, and 28 postinfection. Positive X-gal staining was also observed in the tumor nodule which was challenged with this viral vector. CONCLUSIONS: We have demonstrated that hrR3 is capable of inhibiting Y79 tumor growth both in cell culture and in nude mice. These data suggest that gene therapy using this mutant HSV vector can be a new supplementary therapeutic modality for retinoblastoma. PMID- 10372994 TI - A thin glycoprotein coating of a synthetic lenticule does not cause nutritional deficiency of the anterior cornea. AB - PURPOSE: This study investigated whether a glycoprotein coating that will be used to enhance corneal epithelialization affects in situ nutritional passage through a permeable membrane. METHODS: Sixteen adult cats were equally divided into two groups. Polycarbonate membranes with pore size of 0.1 microm and total pore area (porosity) of 3.1% were used as implant materials. The membranes for Group 1 were coated with a thin layer of Collagen I, while the membranes for Group 2 were uncoated. Each membrane with 8-mm diameter was implanted into an interlamellar pocket of the cornea. The eyes were observed for approximately 35 days to monitor clinical signs of nutritional deficiency of the cornea, and then 7 membranes were removed from the eyes. The permeability of the explanted membranes to glucose, inulin and albumin was used to predict the in situ difference between the coated and uncoated groups in regard to nutritional passage through the membranes. To investigate the long-term effect of the surface coating on corneal health, two animals from Group 1 were followed for up to two years and then both eyes of each animal underwent histological examination. RESULTS: Clinically, no post-surgical complications associated with nutritional deficiency were observed in any of the eyes. Nutritional permeability tests showed no significant differences between the coated and uncoated membranes. Histologically, the long-term animals showed no abnormal morphology associated with nutritional deficiency in the cornea anterior or posterior to the membranes. CONCLUSIONS: A thin glycoprotein coating on a permeable membrane does not appear to affect the nutritional supply of the anterior cornea and therefore can be used to enhance epithelialization of synthetic corneal onlays in vivo. PMID- 10372993 TI - Copurification of selected glycolytic enzymes with retinal S-antigen (arrestin) by hydroxyapatite agarose chromatography of bovine retina. AB - PURPOSE: A variety of methods have been developed for the purification of S antigen but a simple and rapid procedure based on hydroxyapatite-agarose (HA) adsorbtion is most widely used. In the present study, we investigated the nature of proteins purified with the aid of HA chromatography. METHODS: After elimination of retinal S-antigen by HA, the soluble extract of retinal tissue was readsorbed on HA. The proteins were thereafter desorbed by 10 to 500 mM phosphate buffer gradient. Two peaks obtained by SDS-PAGE were used for the generation of specific antisera for subsequent analysis by ELISA and Western blotting. RESULTS: Four proteins (two 48 kDa , one 50 kDa and one 46 kDa) were obtained in this manner. Partial amino acid sequencing permitted the identification of these proteins as alpha-enolase (48 kDa), gamma-enolase (48 kDa), Glucose-6-phosphate Isomerase (50 kDa) and aspartate-amino-transferase (46 kDa). CONCLUSION: The selected glycolytic enzymes co-purified with retinal S-antigen by hydroxyapatite agarose chromatography of bovine retina. PMID- 10372995 TI - Intravitreally injected platelet activating factor induces retinitis in experimental animals. AB - PURPOSE: Platelet activating factor is a lipid which has been strongly implicated in anterior uveitis. In order to investigate further the role of platelet activating factor in intraocular inflammation, we have characterized the histological changes associated with the intravitreal injection of platelet activating factor, PAF analogs, or lyso-PAF in laboratory rabbits and rats. METHODS: Initial studies utilized a PAF analog (rac 1-0-octadecyl 2-0-ethyl glycero phosphoryl choline or ethoxy PAF), because this compound is relatively resistant to degradation by hydrolase, the major degradative enzyme for PAF. Doses ranging from 1 ug to 5 mg and time points from 6 hours to 7 days after injection were studied. RESULTS: In either rats or rabbits, 100 ug of ethoxy PAF consistently induced a marked uveitis with the predominance of inflammation focused in the retina and choroid. Retinitis was also induced in rabbits by either 1 mg PAF injected intravitreally or a similar dose of the PAF precursor/metabolite, lyso PAF. Retinal inflammation was not induced by an inactive lipid, 1,1-0,0-dihexadecyl-rac-glycero-3-phosphocholine, although this compound resulted in mild vitreous inflammation. The histological changes induced by PAF could be readily distinguished from the predominantly anterior inflammation induced by intravitreal injections of substances such an interleukin 1 or endotoxin. CONCLUSIONS: Recent studies indicating that PAF antagonists inhibit a variety of retinal toxicities and our own observations suggest that PAF could be a major mediator of retinal inflammation. PMID- 10372996 TI - IL-4 potentiates IL-1beta- and TNF-alpha-stimulated IL-8 and MCP-1 protein production in human retinal pigment epithelial cells. AB - PURPOSE: Human retinal pigment epithelial (HRPE) cells are involved in ocular inflammation by secretion of chemokines such as IL-8 and MCP-1. It has been shown in this and other laboratories that interleukin-1 beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) are potent inducers of HRPE IL-8 and MCP-1 secretion. The induced IL-8 and MCP-1 expression is often modulated by other proinflammatory factors in a synergistic manner. Modulation of IL-8 and MCP-1 production by interleukin-4 (IL-4), a important mediator in Th2-mediated immunity, and granulocyte/macrophage-colony-stimulating factor (GM-CSF), one of the cytokines secreted by HRPE has been reported in non-ocular cells. The aim of the present investigation was to study effects of these two cytokines alone or in combination with IL-1beta or TNF-alpha on HRPE IL-8 and MCP-1 generation. METHODS: The primary culture of HRPE cells was stimulated with various doses of IL-4, GM-CSF, IL-1beta and TNF-alpha alone or in combination for 8 or 24 hr. The supernatants were subjected to enzyme-linked immunosorbent assay (ELISA) for IL-8 and MCP-1. The mRNAs were isolated from the corresponding cells for Northern blot analysis. RESULTS: IL-1beta and TNF-alpha induced dose-dependent increases in HRPE IL-8 and MCP-1 secretion with maximal stimulation observed at 2-5 ng/ml. IL 4 alone (100 ng/ml) resulted in a slight increase of MCP-1 and IL-8 secretion. When IL-4 was co-administrated with IL-1beta or TNF-alpha, two to three-fold increases in IL-8 and MCP-1 were observed over the maximal levels induced by IL 1beta or TNF-alpha alone. Northern blot analyses revealed that IL-4 did not alter the steady-state MCP-1 mRNA stimulated by IL-1beta and TNF-alpha, or alter the IL 8 mRNA stimulated by TNF-alpha, although the IL-1beta-induced IL-8 mRNA was slightly enhanced by higher concentrations of IL-4 (100 ng/ml). CONCLUSION: The synergistic action by IL-4 occurs predominately at the post-transcriptional level. In contrast to IL-4, GM-CSF alone or in combination with IL-1beta or TNF alpha did not generate additional secretion of HRPE IL-8 and MCP-1. HRPE IL-8 and MCP-1 gene expression and protein production are stimulated by IL-1beta or TNF alpha through pathways differentially modulated by IL-4 and GM-CSF. PMID- 10372997 TI - Clarithromycin for experimental Staphylococcus aureus keratitis. AB - PURPOSE: Clarithromycin, a macrolide antibiotic not previously tested against the common causes of bacterial keratitis, was analyzed for its effectiveness in reducing the number of viable bacteria in a Staphylococcus keratitis model. An in vivo comparison of the effectiveness of clarithromycin to erythromycin, minocycline, and tetracycline for three strains of Staphylococcus aureus was done. METHODS: Rabbit eyes were intrastromally injected with 100 colony forming units of one of three strains of S. aureus. Two strains were methicillin sensitive (ATCC 25923 and MSSA 309) and one strain methicillin-resistant (COL). Eyes were treated every 30 minutes with 0.3% clarithromycin, erythromycin, tetracycline, or minocycline from 4 to 9 hours postinfection. The number of colony forming units (CFU) per cornea in all eyes was determined at 10 hours postinfection. RESULTS: Vehicle-treated and untreated eyes (controls) contained over 6 logs of CFU per cornea, a value significantly higher than any of the antibiotic-treated eyes (P < or = 0.0001). Clarithromycin or erythromycin therapy significantly decreased the number of CFU per cornea by approximately 5 logs in the eyes infected with the methicillin-sensitive strains and by approximately 4 logs in the eyes infected with the methicillin-resistant strain. Tetracycline and minocycline were also successful in treating these strains, but overall showed less effectiveness than clarithromycin and erythromycin. CONCLUSIONS: Clarithromycin proved to be an effective ocular medication for the therapy of experimental S. aureus keratitis. The effectiveness of clarithromycin in this model and its known effectiveness for a variety of bacterial pathogens suggests a role for this drug as a useful ocular antibiotic. PMID- 10372998 TI - Lidocaine toxicity to rat retinal ganglion cells. AB - PURPOSE: To examine the effects of the local anesthetic, lidocaine, on rat retinal ganglion cells (RGC) in vitro and in a modified in vivo assay. METHODS: For in vitro experiments, RGC were dissociated from freshly harvested Long Evan's rat pup retinas. The RGC were incubated overnight with varying concentrations of lidocaine (0.5-12.0 mM). Surviving cells were assayed at 24 hours. In an in vivo assay, 7-day-old Long-Evans rat pups were anesthetized and 2 microl of lidocaine (final intraocular concentration: 0.03-15 mM) or vehicle was injected intravitreally. Intravitreal coinjection of nimodipine or MK801 (dizocilpine) were also performed in a subset of animals. A week after injection, rat pups were sacrificed and each retina removed, dissociated and plated separately. RGC survival was immediately assessed. Living RGC were identified on the basis of morphology and counted in a masked fashion. RESULTS: Lidocaine is toxic in a dose dependent fashion to RGC in vitro. Lower concentrations (0.5 mM and 1.0 mM) were non-toxic; 2.0, 6.0 and 12.0 mM lidocaine killed 25%, 88% and 99% of the RGC respectively. Intravitreal lidocaine was also toxic to RGC in a dose dependent fashion. Lidocaine concentrations of 3.0 mM, 7.5 mM and 15 mM killed 25%, 38% and 44% of the RGC. This effect was blocked by the simultaneous administration of either nimodipine or MK801. CONCLUSIONS: Lidocaine is toxic to RGC both in vitro and in vivo. This effect is blocked in vivo by the simultaneous administration of agents known to block glutamate mediated neuronal death, suggesting that excitotoxicity may be involved in this process. PMID- 10372999 TI - Factors affecting the central corneal thickness of Hong Kong-Chinese. AB - PURPOSE: The aims of this study were to investigate the effect of age, intraocular pressure, refractive error (spherical equivalent) and corneal curvatures on the central corneal thickness of Hong Kong-Chinese. We also compared the central corneal thickness of Hong Kong-Chinese with those previously reported for other national/ethnic groups. METHODS: The central corneal thicknesses of 151 subjects of age 10-60 yrs were measured using an ultrasound pachometer. Intraocular pressure, refractive error and the corneal curvatures of these subjects were also recorded. RESULTS: The mean +/- SD central corneal thickness of the right eye and left eye were 575 +/- 32 microm and 574 +/- 31 microm respectively. No significant difference in central corneal thickness was found between the right and left eyes or between male and female subjects. Central corneal thickness decreased with increasing age but the effect appeared to be due to differences in female subjects only. The maximum decrease in central corneal thickness occurred in subjects in the age group of 10-25 yrs, and in this age group, central corneal thickness and age was significantly correlated in both male and female subjects. Intraocular pressure and central corneal thickness was significantly correlated. There was no correlation between central corneal thickness and refractive error or between central corneal thickness and corneal curvatures. CONCLUSIONS: Central corneal thickness decreased with increasing age but this appeared to be due to female subjects only. Central corneal thickness was significantly correlated with intraocular pressure, but not with refractive error or corneal curvatures. Our subjects also had significantly thicker corneas than those reported for Caucasian subjects. PMID- 10373000 TI - Localization of nitric oxide synthase isoforms in porcine ocular tissues. AB - PURPOSE: To determine by immunohistochemistry the distribution of different nitric oxide synthases (NOS) isoforms in the porcine eye. METHODS: By light microscopy (immunofluorescence), porcine ocular tissues were investigated using monoclonal antibodies against neuronal NOS (nNOS; NOS I), endothelial NOS (ecNOS; NOS III), and macrophage NOS (macNOS; NOS II). RESULTS: Specific nNOS immunoreactivity was detected along the apical cytoplasmic portions of the non pigmented and pigmented ciliary epithelial cells, in the endothelial lining of the corneoscleral meshwork and the uveal cords of the iridocorneal angle tissue, as well as in the photoreceptor layer of the sensory retina. Immunoreactivity for ecNOS was confined to the vascular endothelium of the vessels of the conjunctiva, iris, ciliary body, retina, choroid and optic nerve. A mild immunostaining for macNOS was present in the cytoplasm of some non-pigmented ciliary epithelial cells. CONCLUSIONS: The predominant localization of nNOS in ciliary epithelial and trabecular endothelial cells suggests a possible involvement of nNOS in both the production and outflow of aqueous humor in porcine eyes. PMID- 10373001 TI - Effects of topical carteolol and timolol on tissue circulation in the iris and choroid. AB - PURPOSE: To evaluate the effects of topical carteolol or timolol on tissue circulation in the iris and posterior choroid. METHODS: After a topical instillation of 20 microl of 2% carteolol, 0.5% timolol, or physiological saline (for control) into one eye, and physiological saline into the other eye of pentobarbital-anesthetized Dutch pigmented rabbits, normalized blur value; a quantitative index of tissue blood velocity in the iris (NB(iris)) and posterior choroid (NB(cho)) was obtained using the laser-speckle method. Intraocular pressure (IOP), blood pressure and pulse rate were serially monitored for 2 hours after instillation. Using separate groups of rabbits, NB(iris) and IOP were measured before and after 20-day twice-daily unilateral treatment of carteolol or timolol. RESULTS: After a single instillation of carteolol, NB(iris) was significantly greater only in the treated eyes than control eyes (P = 0.0050, repeated measures two-way ANOVA), while NB(cho) showed no significant change. IOP in the treated eyes significantly reduced (P = 0.0005). Bilateral reductions of tissue vascular resistance in the iris were found after carteolol instillation (P = 0.0183 approximately 0.0322). After timolol instillation, serial changes in NB(iris) and NB(cho) in the treated eyes were significantly different from those in control eyes (P = 0.0129, 0.0031), while there were no significant differences at any of time points (Mann-Whitney test); IOP in both eyes was significantly reduced (P = 0.0096 approximately 0.0005); tissue vascular resistance in the iris and posterior choroid showed no significant changes. After 20-day treatment, NB(iris) in the both eyes of carteolol-treated rabbits significantly increased from the baseline (P = 0.0280, 0.0425, Wilcoxon signed rank test) and NB(iris) in timolol-treated eyes significantly decreased (P = 0.0280). CONCLUSIONS: A single instillation of topical carteolol significantly increased the iris tissue blood velocity in the treated eye and reduced the tissue vascular resistance in both eyes. Topical timolol tended to decrease tissue blood velocity in the iris and choroid of the treated eye, but showed no significant effects on tissue vascular resistance in the both tissues. PMID- 10373002 TI - Flexible ligand docking: a multistep strategy approach. AB - A flexible ligand docking protocol based on a divide-and-conquer strategy is investigated. This approach first separates total search space into conformation and orientation space. It uses a grid-based method to sample the conformation of an unbound ligand and to select the low-energy conformers. Rigid docking is then carried out to locate the low-energy binding orientations for these conformers. These docking structures are subsequently subjected to structure refinement including molecular mechanics minimization, conformational scanning at the binding site and a short period of molecular dynamics-based simulated annealing. This approach has been applied to twelve ligand-protein complexes with three to sixteen rotatable bonds. The docked lowest-energy structures have root mean square deviations ranging from 0.64 A to 2.01 A with respect to the corresponding crystal structures. The effect of atomic charges and van der Waals parameters on the docking results, and the role of the dielectric constant in the conformation sampling are discussed in detail. A fragment-based docking approach that takes advantages of the divide-and-conquer strategy has also been explored and the results are compared with those produced by a whole molecule-based approach. PMID- 10373003 TI - The Helicobacter pylori genome: from sequence analysis to structural and functional predictions. AB - Fold assignments for proteins from the Helicobacter pylori genome are carried out using BASIC, a profile-profile alignment algorithm recently tested on the Mycoplasma genitalium and Escherichia coli genomes. The fold assignments are followed by automated function evaluation, based on the multilevel description of functional sites in proteins. Over 40% of the proteins encoded in the H. pylori genome can be recognized as belonging to a protein family with known structure. Previous estimates suggested that only 10-15% of genome proteins could be characterized this way. This dramatic increase in the number of recognized homologies between H. pylori proteins and structurally characterized protein families is partly due to the rapid increase of the database of known protein structures, but mostly it is due to the significant improvement in prediction algorithms. Knowledge of a protein fold adds a new dimension to our understanding of its function and, similarly, structure prediction can also add to understanding, verification, and/or prediction of function for uncharacterized proteins. Several examples analyzed in more detail in this article illustrate insights that can be achieved from structure and detailed function prediction. PMID- 10373004 TI - Serpins in the Caenorhabditis elegans genome. AB - Data mining in genome sequences can identify distant homologues of known protein families, and is most powerful if solved structures are available to reveal the three-dimensional implications of very dissimilar sequences. Here we describe putative serpin sequences identified with very high statistical significance in the Caenorhabditis elegans genome. When mapped onto vertebrate serpins such as alpha1-antitrypsin, they suggest novel structural features. Some appear complete, some show extensive deletions, and others appear to contain only the C-terminal part of the known serpin fold, probably in partnership with N-terminal regions that have conformations unlike those of known serpins. The observation of such striking sequence similarity, in proteins that must have significantly different overall structures, substantially extends the structural characteristics of the serpin family of proteins. PMID- 10373005 TI - Time resolved fluorescence and phosphorescence properties of the individual tryptophan residues of barnase: evidence for protein-protein interactions. AB - Steady-state and time-resolved fluorescence, as well as phosphorescence measurements, were used to resolve the luminescence properties of the three individual tryptophan residues of barnase. Assignment of the fluorescence properties was performed using single-tryptophan-containing mutants and the results were compared with the information available from the study of wild-type and two-tryptophan-containing mutants (Willaert, Lowenthal, Sancho, Froeyen, Fersht, Engelborghs, Biochemistry 1992;31:711-716). The fluorescence and the phosphorescence emission of wild-type barnase is dominated by Trp35, although Trp71 has the strongest intrinsic fluorescence when present alone. Fluorescence emission of these two tryptophan residues is blue-shifted and pH-independent. The fluorescence decay parameters of Trp94 are pH-dependent, and an intramolecular collision frequency of 2 to 5 x 10(9) s(-1) between Trp94 and His18 is calculated. Fluorescence emission of Trp94 is red-shifted. Fluorescence anisotropy decay reveals the local mobility of the individual tryptophan residues and this result correlates well with their phosphorescence properties. Trp35 and Trp71 display a single phosphorescence lifetime, which reflects the rigidity of their environment. Surface Trp94 does not exhibit detectable phosphorescence emission. The existence of energy transfer between Trp71 and Trp94, as previously detected by fluorescence measurements, is also observed in the phosphorescence emission of barnase. Dynamic quenching causes the phosphorescence intensity to be protein-concentration dependent. In addition, fluorescence anisotropy shows concentration dependency, and this can be described by the formation of trimers in solution. PMID- 10373006 TI - Knowledge-based interaction potentials for proteins. AB - We discuss the derivation of atomic-level potentials of mean force from the known protein structures and their applicability for structural evaluation applications. In the derivation process, rigorous density estimation methodology is used to estimate the probability density functions (PDFs) for the distributions of interatomic distances in the protein structures. Potentials of mean force are then derived from these density functions using simple Boltzmann's relation. We also test the potentials against pairs of current and superseded protein structures in the Protein Data Bank. Using PDF potentials to evaluate each structure pair, we are able to identify, with high accuracy, which of the two structures is of higher resolution or better quality. This result shows that the PDF potentials are sensitive to details in protein structures as the current and superseded atomic coordinates generally do not differ by more than 1 A in root-mean-square deviation, and that the PDF potentials could potentially be used for X-ray structure refinement and protein structure prediction. PMID- 10373007 TI - Hidden Markov models that use predicted secondary structures for fold recognition. AB - There are many proteins that share the same fold but have no clear sequence similarity. To predict the structure of these proteins, so called "protein fold recognition methods" have been developed. During the last few years, improvements of protein fold recognition methods have been achieved through the use of predicted secondary structures (Rice and Eisenberg, J Mol Biol 1997;267:1026 1038), as well as by using multiple sequence alignments in the form of hidden Markov models (HMM) (Karplus et al., Proteins Suppl 1997;1:134-139). To test the performance of different fold recognition methods, we have developed a rigorous benchmark where representatives for all proteins of known structure are matched against each other. Using this benchmark, we have compared the performance of automatically-created hidden Markov models with standard-sequence-search methods. Further, we combine the use of predicted secondary structures and multiple sequence alignments into a combined method that performs better than methods that do not use this combination of information. Using only single sequences, the correct fold of a protein was detected for 10% of the test cases in our benchmark. Including multiple sequence information increased this number to 16%, and when predicted secondary structure information was included as well, the fold was correctly identified in 20% of the cases. Moreover, if the correct secondary structure was used, 27% of the proteins could be correctly matched to a fold. For comparison, blast2, fasta, and ssearch identifies the fold correctly in 13-17% of the cases. Thus, standard pairwise sequence search methods perform almost as well as hidden Markov models in our benchmark. This is probably because the automatically-created multiple sequence alignments used in this study do not contain enough diversity and because the current generation of hidden Markov models do not perform very well when built from a few sequences. PMID- 10373008 TI - Molecular dynamics simulations of human alpha-lactalbumin: changes to the structural and dynamical properties of the protein at low pH. AB - Two 700-ps molecular dynamics simulations of human alpha-lactalbumin have been compared. Both were initiated from an X-ray structure determined at pH 6.5. One simulation was designed to represent native conditions and the other the protein in solution at pH 2.0 without a bound calcium ion. The low pH conditions were modelled by protonating the aspartate, glutamate, and histidine side chains and the protein C-terminus. Significant changes were observed for the C-terminal region of the sequence in the simulation at low pH. Most notably an alpha-helix, helix D, and the C-terminal 3(10) helix were substantially disrupted relative to the simulation at high pH. These perturbations to the native fold are similar to those observed in an X-ray structure of alpha-lactalbumin at pH 4.2. In addition, larger fluctuations about side chain torsion angles were observed in the low pH simulation than in that corresponding to the higher pH. These structural and dynamical changes might be representative of the early stages of the transition to the molten-globule state of the protein known to be formed under low pH conditions in solution. PMID- 10373009 TI - Exploring the dynamic information content of a protein NMR structure: comparison of a molecular dynamics simulation with the NMR and X-ray structures of Escherichia coli ribonuclease HI. AB - The multiconformer nature of solution nuclear magnetic resonance (NMR) structures of proteins results from the effects of intramolecular dynamics, spin diffusion and an uneven distribution of structural restraints throughout the molecule. A delineation of the former from the latter two contributions is attempted in this work for an ensemble of 15 NMR structures of the protein Escherichia coli ribonuclease HI (RNase HI). Exploration of the dynamic information content of the NMR ensemble is carried out through correlation with data from two crystal structures and a 1.7-ns molecular dynamics (MD) trajectory of RNase HI in explicit solvent. Assessment of the consistency of the crystal and mean MD structures with nuclear Overhauser effect (NOE) data showed that the NMR ensemble is overall more compatible with the high-resolution (1.48 A) crystal structure than with either the lower-resolution (2.05 A) crystal structure or the MD simulation. Furthermore, the NMR ensemble is found to span more conformational space than the MD simulation for both the backbone and the sidechains of RNase HI. Nonetheless, the backbone conformational variability of both the NMR ensemble and the simulation is especially consistent with NMR relaxation measurements of two loop regions that are putative sites of substrate recognition. Plausible side chain dynamic information is extracted from the NMR ensemble on the basis of (i) rotamericity and syn-pentane character of variable torsion angles, (ii) comparison of the magnitude of atomic mean-square fluctuations (msf) with those deduced from crystallographic thermal factors, and (iii) comparison of torsion angle conformational behavior in the NMR ensemble and the simulation. Several heterogeneous torsion angles, while adopting non-rotameric/syn-pentane conformations in the NMR ensemble, exist in a unique conformation in the simulation and display low X-ray thermal factors. These torsions are identified as sites whose variability is likely to be an artifact of the NMR structure determination procedure. A number of other torsions show a close correspondence between the conformations sampled in the NMR and MD ensembles, as well as significant correlations among crystallographic thermal factors and atomic msf calculated from the NMR ensemble and the simulation. These results indicate that a significant amount of dynamic information is contained in the NMR ensemble. The relevance of the present findings for the biological function of RNase HI, protein recognition studies, and previous investigations of the motional content of protein NMR structures are discussed. PMID- 10373010 TI - A preliminary CD and NMR study of the Escherichia coli DNA polymerase III theta subunit. AB - The theta subunit of DNA polymerase III, the main replicative polymerase of Escherichia coli, has been examined by circular dichroism and by NMR spectroscopy. The polymerase core consists of three subunits: alpha, epsilon, and theta, with alpha possessing the polymerase activity, epsilon functioning as a proofreading exonuclease, and theta, a small subunit of 8.9 kD, of undetermined function. The theta subunit has been expressed in E. coli, and a CD analysis of theta indicates the presence of a significant amount of secondary structure: approximately 52% alpha helix, 9% beta sheet, 21% turns, and 18% random coil. However, at higher concentrations, theta yields a poorly-resolved 1D proton NMR spectrum in which both the amide protons and the methyl protons show poor chemical shift dispersion. Subsequent 1H-15N HSQC analysis of uniformly-15N labeled theta supports the conclusion that approximately half of the protein is reasonably well-structured. Another quarter of the protein, probably including some of the N-terminal region, is highly mobile, exhibiting a chemical shift pattern indicative of random coil structure. The remaining amide resonances exhibit significant broadening, indicative of intermolecular and/or intramolecular exchange processes. Improved chemical shift dispersion and greater uniformity of resonance intensities in the 1H-15N HSQC spectra resulted when [U 15N]-theta was examined in the presence of epsilon186--the N-terminal domain of the epsilon-subunit. Further work is currently in progress to define the solution structure of theta and the theta-epsilon186 complex. PMID- 10373011 TI - Conserved water molecules in a large family of microbial ribonucleases. AB - We systematically analyzed the crystallographically determined water molecules of all known structures of RNase T1 and compared them to the ordered solvent in a large number of related microbial nucleases. To assess the crystallographers' impact on the interpretation of the solvent structure, we independently refined five validation structures from diffraction data derived from five isomorphous crystals of RNase T1. We also compared the positions of water molecules found in 11 published isomorphous RNase T1 inhibitor complexes. These data suggest that the positions of most of the waters located on the surface of a protein and that are well-determined in the experimental electron density maps are determined primarily by crystal packing forces. Water molecules with less well-defined electron density are in general unique to one or a small number of crystal structures. Only a small number of the well-defined waters are found to be independent of the crystal environment. These waters have a low accessible surface area and B-factor, and tend to be conserved in the crystal structures of a number of evolutionary related ribonucleases as well. A single water molecule is found conserved in all known microbial ribonucleases. PMID- 10373013 TI - Immunolocalization of mitsugumin29 in developing skeletal muscle and effects of the protein expressed in amphibian embryonic cells. AB - The temporal appearance and subcellular distribution of mitsugumin29 (MG29), a 29 kDa transmembrane protein isolated from the triad junction in skeletal muscle, were examined by immunohistochemistry during the development of rabbit skeletal muscle. MG29 appeared in the sarcoplasmic reticulum (SR) in muscle cells at fetal day 15 before the onset of transverse tubule (T tubule) formation. In muscle cells at fetal day 27, in which T tubule and triad formation is ongoing, both SR and triad were labeled for MG29. In muscle cells at newborn 1 day, the labeling of the SR had become weak and the triads were well developed and clearly labeled for MG29. Specific and clear labeling for MG29 was restricted to the triads in adult skeletal muscle cells. When MG29 was expressed in amphibian embryonic cells by injection of the cRNA, a large quantity of tubular smooth-surfaced endoplasmic reticulum (sER) was formed in the cytoplasm. The tubular sER was 20-40 nm in diameter and appeared straight or reticular in shape. The tubular sER was formed by the fusion of coated vesicles [budded off from the rough-surfaced endoplasmic reticulum (rER)] and vacuoles of rER origin. The present results suggest that MG29 may play important roles both in the formation of the SR and the construction of the triads during the early development of skeletal muscle cells. PMID- 10373014 TI - RARbeta mediates the response of Hoxd4 and Hoxb4 to exogenous retinoic acid. AB - One action of retinoids in development is the regulation of Hox gene expression. Hoxd4 and Hoxb4 expression in the embryonic hindbrain is anteriorized by retinoic acid (RA) treatment of mid-gestation mouse embryos. Here we demonstrate that retinoic acid receptor beta (Rarb) deficient mice present only a partial anteriorization of either Hoxd4 or Hoxb4 in response to RA treatment. Our results strongly suggest that RARbeta is a mediator of their RA-response, and reveal anteroposterior polarity within a single rhombomere (r). Additionally, we generated mice doubly mutated for Hoxd4 and Rarb in an attempt to identify common morphogenetic pathways between these two genes. We conclude that there are no synergistic interactions between Hoxd4 and Rarb in the specification of the cervical vertebrae. PMID- 10373015 TI - Expression and genetic analysis of prtb, a gene that encodes a highly conserved proline-rich protein expressed in the brain. AB - A mouse gene, designated prtb (proline codon-rich transcript, brain expressed) was identified and characterized from a gene trap embryonic stem cell line. It encodes a proline-rich protein of 168 amino acids that shares 99% amino acid sequence identity with its human homologue and is located on the distal region of mouse chromosome 15. To determine the expression pattern and function of prtb, mice that carry the prtb(gt) allele were generated. During embryogenesis,prtb gene expression as revealed by beta-galactosidase (beta-gal) marker gene activity was highly regulated. Between embryonic day (E) 11.5 and E12.5, beta-gal activity was restricted to the developing heart. From E13.5 on, expression in the heart was extinguished. However, very strong beta-gal activity could be detected in the brains of adult mice, suggesting a role for this gene in brain function. Mice homozygous for the mutation were viable, fertile, and did not display any obvious abnormalities. This could be due to functional redundancy as Northern blot hybridization analysis clearly demonstrated that prtb(gt) is likely to be a null allele. PMID- 10373012 TI - Are membrane proteins "inside-out" proteins? AB - One of the central paradigms of structural biology is that membrane proteins are "inside-out" proteins, in that they have a core of polar residues surrounded by apolar residues. This is the reverse of the characteristics found in water soluble proteins. We have decided to test this paradigm, now that sufficient numbers of transmembrane alpha-helical structures are accessible to statistical analysis. We have analyzed the correlation between accessibility and hydrophobicity of both individual residues and complete helices. Our analyses reveal that hydrophobicity of residues in a transmembrane helical bundle does not correlate with any preferred location and that the hydrophilic vector of a helix is a poor indicator of the solvent exposed face of a helix. Neither polar nor hydrophobic residues show any bias for the exterior or the interior of a transmembrane domain. As a control, analysis of water-soluble helical bundles performed in a similar manner has yielded clear correlations between hydrophobicity and accessibility. We therefore conclude that, based on the data set used, membrane proteins as "inside-out" proteins is an unfounded notion, suggesting that packing of alpha-helices in membranes is better understood by maximization of van der Waal's forces, rather than by a general segregation of hydrophobicities driven by lipid exclusion. PMID- 10373017 TI - DNA methylation pattern of a tandemly repeated LacZ transgene indicates that most copies are silent. AB - The Pgk-1,2-lacZ transgene consists of the ubiquitously-expressed Pgk-1 promoter driving expression of the E. coli lacZ reporter gene. We studied the expression of this transgene in a mouse strain carrying 8-9 tandem copies of this construct. When inherited through the male germ line, the transgene was expressed in all tissues examined but when inherited through the female germ line, the transgene became irreversibly inactivated. The lacZ region is a CpG-rich island that was essentially entirely methylated in all copies of the silent, maternally-inherited transgene. At the active transgenic locus, all but one of the copies were entirely methylated. This one unmethylated copy was adjacent to the cellular DNA and was presumed to be the expressed transgene copy. These results suggest that the tandem repeats of transgenes become silenced by a mechanism associated with DNAmethylation and that proximity to the cellular genome may be important in maintaining expression against the spread of inactivation from the adjacent silent transgenes. PMID- 10373016 TI - Facial development and type III collagen RNA expression: concurrent repression in the osteopetrotic (Toothless,tl) rat and rescue after treatment with colony stimulating factor-1. AB - The toothless (osteopetrotic) mutation in the rat is characterized by retarded development of the anterior facial skeleton. Growth of the anterior face in rats occurs at the premaxillary-maxillary suture (PMMS). To identify potential mechanisms for stunted facial growth in this mutation we compared the temporospatial expression of collagen I (Col I) and collagen III (Col III) RNA around this suture in toothless (tl) rats and normal littermates by in situ hybridization of specific riboprobes in sagittal sections of the head. In normal rats, the suture is S shaped at birth and becomes highly convoluted by 10 days with cells in the center (fibroblasts and osteoblast progenitors) expressing Col III RNA and those at the periphery (osteoblasts) expressing no Col III RNA but high amounts of Col I RNA throughout the growth phase (the first 2 postnatal weeks). In the mutant PMMS, cells were reduced in number, less differentiated, and fewer osteoblasts were encountered. Expression of Col I RNA was at normal levels, but centrosutural cells expressed Col III RNA only after day 6 and then only weakly. A highly convoluted sutural shape was never achieved in mutants during the first 2 postnatal weeks. Treatment of tl rats with the cytokine CSF-1 improved facial growth and restored cellular diversity and Col III RNA expression in the PMMS to normal levels. Taken together, these data suggest that normal facial growth in rats is related to expression of Col III RNAby osteoblast precursors in the PMMS, that these cells are deficient in the tl mutation and are rescued following treatment with CSF-1. PMID- 10373018 TI - Detailed characterization of the human aorta-gonad-mesonephros region reveals morphological polarity resembling a hematopoietic stromal layer. AB - The definitive long-term repopulating human hematopoietic stem cell, which seeds the adult blood system, was previously thought to derive from the extra-embryonic yolk sac. However, there is now considerable evidence that in both avian and murine systems, yolk sac hematopoietic cells are largely a transient, embryonic population and the definitive stem cell, in fact, derives from a distinct region within the embryonic mesoderm, the aorta-gonad-mesonephros region. In the human embryo, an analogous region has been found to contain a cluster of cells distinct from, but closely associated with, the ventral endothelium of the dorsal aorta, the appearance of which is restricted both spatially and temporally. We have used antibodies recognising hematopoietic regulatory factors to further characterise this region in the human embryo. These studies indicate that all factors examined, including vascular endothelial growth factor and its receptor FLK-1, Flt-3 ligand and its receptor STK-1, and stem cell leukemia transcription factor, are expressed by both hematopoietic cells in the cluster and endothelial cells. However, there is some discontinuity in cells directly underlying the cluster. Furthermore, we have identified a morphologically distinct region of densely packed, rounded cells in the mesenchyme directly beneath the ventral wall of the dorsal aorta, and running along its entire length. In the preumbilical AGM region, directly underlying the hematopoietic cluster, but not at more rostral and caudal levels, this region of mesenchyme expresses tenascin-C, an extracellular matrix glycoprotein known to facilitate cell-cell interactions and migration. This region of cells may therefore provide the microenvironmental support for the intraembryonic development of definitive hematopoietic stem cells, a process in which tenascin-C may play a pivotal role. PMID- 10373019 TI - EGF-dependent lobule formation and FGF7-dependent stalk elongation in branching morphogenesis of mouse salivary epithelium in vitro. AB - When supplemented with appropriate growth factors, salivary gland epithelial explants isolated from mouse embryos undergo branching morphogenesis in vitro in the absence of mesenchyme. Epidermal growth factor (EGF) induces lobule formation, while fibroblast growth factor 7 (FGF7) promotes stalk elongation. A mixture of EGF and FGF7 produces an intermediate morphology, which resembles the branching pattern of salivary epithelium observed in vivo. To investigate how lobule formation and stalk elongation are related to the pattern of epithelial cell proliferation induced by EGF and FGF7, we performed a bromodeoxyuridine labeling study in whole-mount preparations. During the initial steps of lobule formation in EGF cultures, cleft and non-cleft regions had similar proliferative activity. However, once clefts had fully deepened, cells with low proliferative activity appeared at the bottom of the clefts. In contrast, during stalk elongation in FGF7 cultures, distal regions of the explants always showed higher proliferative activity than proximal regions. These results suggest that stalk elongation, but not cleft formation, may result from differential cell proliferation. PMID- 10373020 TI - Immunocytochemical studies of the interactions of cadherins and catenins in the early Xenopus embryo. AB - Linkage of cadherins to the cytoskeleton is crucial for their adhesive function. Since alpha- and beta-catenin play a key role in this linkage, these proteins are possible targets for processes that control cell-cell adhesion. To achieve a better understanding of the regulation of cell-cell adhesion in embryonic morphogenesis, we used immunohistology to investigate how in Xenopus blastomeres catenins respond to disturbances in the expression of maternal cadherins. Overexpression of myc-tagged maternal cadherin leads to a proportionate increase of the level of beta-catenin. The two proteins colocalize in the endoplasmic reticulum, in cytoplasmic vesicles, and along the cell membrane, indicating that the beta-catenin binds to overexpressed cadherin early in its passage to the plasma membrane. Expression of cadherin is essential for the stable presence of beta-catenin, as depletion from maternal cadherin mRNA leads to a complete loss of beta-catenin from the blastomeres. alpha-Catenin behaves differently. Overexpression of cadherin leaves the amount and localization of alpha-catenin largely unaffected, and additional cadherin inserts itself into the membrane without a proportionate rise in the level of membrane-bound alpha-catenin. However, cadherin mRNA depletion leads to a redistribution of alpha-catenin from the membrane to the cytoplasm. Thus, cadherin is required to localize alpha catenin to the membrane, but the amount of alpha-catenin along the membrane seems to be restricted to a certain level which cannot be exceeded. The relevance of these observations for the regulation of cadherin-mediated cell adhesion in the Xenopus embryo is discussed. Additionally, we demonstrate that plakoglobin, like beta-catenin an armadillo repeat protein, shows neither accumulation after overexpression nor colocalization with the overexpressed cadherin. PMID- 10373022 TI - Virtual MR arthroscopy: new insights into joint morphology. AB - Evaluation of the feasibility of virtual magnetic resonance (MR) arthroscopy was performed on 10 patients referred for MR arthrography of the shoulder, the elbow, or the knee. A fast spoiled contrast-enhanced three-dimensional gradient-echo sequence was combined with image postprocessing to render arthroscopic views. Virtual MR arthroscopy was successfully demonstrated in all 10 joints. Determination of added diagnostic benefit will require further study. PMID- 10373021 TI - Alternative splicing and embryonic expression of the Xenopus mad4 bHLH gene. AB - The cell proliferative activity of the Myc family of basic helix-loop helix/leucine zipper (bHLHZip) transcription factors is dependent upon binding to the ubiquitous Max protein. In the absence of heterodimerization with Max, Myc protein is unable to efficiently bind to DNA and activate transcription. Members of the Mad family of transcription factors are thought to modulate the cell proliferative effects of the c-myc proto-oncogene by binding to Max, directly competing with the Myc protein for both heterodimerization and DNA binding. Consistent with a role in down-regulating cell division, the murine mad genes are expressed in embryonic tissues undergoing differentiation, often during or shortly after the down-regulation of myc gene expression. Here, we report the isolation and characterization of the first Xenopus mad family member, Xmad4. Maternal Xmad4 transcripts are present at high levels in the oocyte and in the cleavage stage embryo, but almost disappear by the neurula stage. Zygotic expression of the Xmad4 gene is initiated in the epidermis of the late neurula stage, and shortly thereafter, Xmad4 is transiently detectable in the cement and hatching glands. At later stages, expression is also observed in the developing pronephros and liver. Unlike the murine mad4 gene, we find that multiple Xmad4 splice variants exist in Xenopus and that these variants are differentially expressed in both the embryo and the adult. Despite the demonstrated antagonistic role of Mad proteins in the regulation of Myc activity, we show that the over expression of Xmad4 in the cleavage-stage embryo has no detectable phenotypic effect, suggesting that Myc function is dispensable during early embryonic development. PMID- 10373023 TI - MR perfusion imaging in human brain using the UNFAIR technique. Un-inverted flow sensitive alternating inversion recovery. AB - Pulsed arterial spin labeling magnetic resonance techniques have been developed recently to estimate cerebral blood flow (CBF). Flow-sensitive alternating inversion recovery (FAIR) is one such technique that has been implemented successfully in humans. Un-inverted FAIR (UNFAIR) is an alternative technique in which the flow-sensitive image is acquired following inversion of all spins outside the slice of interest, and the control image is acquired without any spin labeling. This approach is potentially more efficient than FAIR since the UNFAIR control image is entirely flow independent and need only be acquired once. Here, we describe implementation of the sequence on a clinical 1.5 T magnetic resonance system. Both FAIR and UNFAIR perfusion-weighted images were obtained from six normal volunteers. Wash-in/wash-out curves measured in cortical gray and white matter were practically identical for the two techniques, as predicted by our model. PMID- 10373025 TI - Equilibrium phase MR angiography of the aortic arch and abdominal vasculature with the blood pool contrast agent CMD-A2-Gd-DOTA in pigs. AB - Meglumine-carboxymethyldextran-ethylenediamino-gadoterate (CMD-A2-Gd-DOTA) was evaluated as a blood pool contrast agent for magnetic resonance angiography (MRA). MRA of large body vasculature was performed in seven pigs using a gradient echo sequence at 1.5 T before and after 0.05 mmol/kg CMD-A2-Gd-DOTA injection. The signal- and contrast-to-noise ratios (SNRs, CNRs) were measured, as well as the pharmacokinetic clearance pattern. CMD-A2-Gd-DOTA visualized the vasculature with a high SNR and CNR for over 110 minutes after injection, but for the renal arteries the CNR was only significant within 15 minutes. Image quality was maximum within 15 minutes, producing enhancement (mean +/- SD) as follows: aortic arch 738 +/- 272%, abdominal aorta 393 +/- 123%, left renal artery 202 +/- 95%, right renal artery 248 +/- 107%, inferior vena cava 371 +/- 129%, and portal vein 513 +/- 145%, all P values < or =0.001. The clearance pattern was triphasic. Due to the excellent enhancement of vasculature without background enhancement, CMD A2-Gd-DOTA is potentially a useful MR blood pool contrast agent for equilibrium phase MRA. PMID- 10373024 TI - Fast tissue segmentation based on a 4D feature map in characterization of intracranial lesions. AB - The aim of this work was to develop a fast and accurate method for tissue segmentation in magnetic resonance imaging (MRI) based on a four-dimensional (4D) feature map and compare it with that derived from a 3D feature map. High resolution MRI was performed in 5 normal individuals, in 12 patients with brain multiple sclerosis (MS), and 9 patients with malignant brain tumors. Three inputs (proton-density, T2-weighted fast spin-echo, and T1-weighted spin-echo MR images) were routinely utilized. As a fourth input, either magnetization transfer MRT was used or T1-weighted post-contrast MRI (in patients only). A modified k-nearest neighbor segmentation algorithm was optimized for maximum computation speed and high-quality segmentation. In that regard, we a) discarded the redundant seed points; b) discarded the points within 0.5 standard deviation from the cluster center that were non-overlapping with other tissue; and c) removed outlying seed points outside 5 times the standard deviation from the cluster center of each tissue class. After segmentation, a stack of color-coded segmented images was created. Our new technique utilizing all four MRI inputs provided better segmentation than that based on three inputs (P < 0.001 for MS and P < 0.001 for tumors). The tissues were smoother due to the reduction of statistical noise, and the delineation of the tissues became sharper. Details that were previously blurred or invisible now became apparent. In normal persons a detailed depiction of deep gray matter nuclei was obtained. In malignant tumors, up to five abnormal tissue types were identified: 1) solid tumor core, 2) cyst, 3) edema in white matter 4) edema in gray matter, and 5) necrosis. Delineation of MS plaque in different stages of demyelination became much sharper. In conclusion, the proposed methodology warrants further development and clinical evaluation. PMID- 10373026 TI - Differences between normal subjects and patients with coronary artery disease for three different MR coronary angiography respiratory suppression techniques. AB - A comparison between three magnetic resonance coronary angiography (MRCA) respiratory motion suppression techniques was performed for both normal subjects and patients with coronary artery disease (CAD). MRCA images were acquired in 17 normal subjects and 15 patients with CAD, using conventional breath-hold MRCA, navigator echo (NE)-guided breath-hold MRCA (LED feedback), and NE-gated MRCA during free respiration. Image quality, diaphragm registration, and total acquisition time were assessed. Overall, there was poor diaphragm registration for conventional breath-holding compared with free respiration (P < 0.001). CAD patients found it significantly more difficult to perform a steady breath-hold (P = 0.04) or attain the same diaphragm position over multiple breath-holds than normal subjects (P = 0.02). All normal subjects, but only 3 of the 15 CAD patients, were able to perform the LED feedback technique (P < 0.001). For normal subjects, image quality was similar between the three respiratory suppression techniques (P = 0.3), while for CAD patients there was an improvement in image quality, for images acquired during free respiration (breath-hold vs. free respiration, P < 0.01). There was no significant difference in the total acquisition times between the breath-hold and free respiration techniques (P = 0.2). There were substantial differences in the effectiveness of MRCA respiratory suppression techniques between normal subjects and CAD patients. In patients, only NE-gated MRCA performed well, requiring minimal cooperation with no increase in total acquisition time. Validation of NE-MRCA techniques should always be performed in patients, as well as normal subjects, to ensure correct evaluation of the technique for the target population. PMID- 10373027 TI - Spatial and temporal modulation of perfusion in the rat ovary measured by arterial spin labeling MRI. AB - The hemodynamic changes triggered by luteinizing hormone/human chorionic gonadotropin (LH/hCG) in ovaries of immature pregnant mare serum gonadotropins (PMSG)-primed female Wistar rats were followed by pulsed arterial spin labeling magnetic resonance imaging. Decreased perfusion was monitored in the first 2 hours after administration of hCG followed by a transient significant rise in perfusion. Subsequently, constant ovarian perfusion of 10.9 +/- 4.3 mL min(-1) g( 1) was maintained during the exponential increase in ovarian volume. However, ovarian perfusion was not uniform, and prior to ovulation poorly perfused regions were detected that were assigned to the follicular fluid in preovulatory follicles. This result implied that in the time scale of seconds, corresponding to the T1 relaxation time of water in the follicular fluid, exchange of arterial water with water in the follicular fluid was negligible. Along with the drop in the levels of high-energy phosphate metabolites detected by 31P nuclear magnetic resonance spectroscopy and the shift to glycolytic metabolism, these results support the hypothesis that physiological hypoxia could play a role in large preovulatory follicles as part of the normal ovarian cycle. PMID- 10373028 TI - Orientation-independent diffusion imaging without tensor diagonalization: anisotropy definitions based on physical attributes of the diffusion ellipsoid. AB - Diffusion tensor imaging can provide a complete description of the diffusion process in tissue. However, this description is not unique but is orientation dependent, and, to quantify properly the intrinsic orientation-independent diffusion properties of the tissue, a set of three rotationally invariant quantities is needed. Instead of using the tensor eigenvalues for this, we define a new set consisting of scaled invariants that have the proper magnitude of actual diffusion constants and that are directly related to the physical attributes of the diffusion ellipsoid, namely, its average radius, surface, and volume. Using these three physical invariants, a new family of anisotropy measures is defined that are normalized between zero (isotropic) and one (completely anisotropic). Because rotational invariants are used, this approach does not require tensor diagonalization and eigenvalue determination and is therefore not susceptible to potential artifacts induced during these number manipulations. The relationship between the new anisotropy definitions and existing orientation-independent anisotropy indices obtained from eigenvalues is discussed, after which the new approach is evaluated for a group of healthy volunteers. PMID- 10373029 TI - Tracking oxygen effects on MR signal in blood and skeletal muscle during hyperoxia exposure. AB - Blood and muscle T1 and T2 relaxivity was examined under normoxic (air; 20.8% O2) and hyperoxic (100% O2) conditions to determine whether the oxygenation state of blood in the large vessels and in the microcirculation can be monitored in vivo. The femoral artery/vein and the soleus and gastrocnemius muscles were examined in healthy human male volunteers. Arterial blood T1 decreased with hyperoxia, while venous blood T2 increased, due to increased dissolved O2 and decreased deoxyhemoglobin, respectively. A biexponential T2 model of muscle is proposed, where the short T2 component reflects primarily the intracellular and interstitial compartments (in fast exchange), and the long T2 reflects blood. In this model, the long T2 component increased with hyperoxia exposure. This was more evident in slow twitch (soleus) than in fast twitch (gastrocnemius) muscle. It is concluded that changes in the long T2 component reflect change in the microcirculation oxygenation state. PMID- 10373030 TI - MRI geometric distortion: a simple approach to correcting the effects of non linear gradient fields. AB - We present a method to correct intensity variations and voxel shifts caused by non-linear gradient fields in magnetic resonance images. The principal sources of distortion are briefly discussed, as well as the methods of correction currently in use. The implication of the gradient field non-linearities on the signal equations are described in a detailed way for the case of two- and three dimensional Fourier imaging. A model of these non-linearities, derived from the geometry of the gradient coils, is proposed and then applied in post-processing to correct any images regardless of the acquisition sequence. Initial position errors, as large as 4 mm (i.e., four voxels of 1 x 1 x 1.4 mm3) before correction, are reduced to less than the voxel sizes after correction. PMID- 10373031 TI - Measurement of the apparent diffusion coefficient in diffuse renal disease by diffusion-weighted echo-planar MR imaging. AB - The purpose of this study was to determine the relationship between the apparent diffusion coefficient (ADC) and diffuse renal disease by diffusion-weighted echolanar magnetic resonance (MR) imaging (EPI). Thirty-four patients were examined with diffusion-weighted EPI. The average ADC values were 2.55 x 10(-3) mm2/sec for the cortex and 2.84 x 10(-3) mm2/sec for the medulla in the normal kidneys. The ADC values in both the cortex and medulla in chronic renal failure (CRF) kidneys and in acute renal failure (ARF) kidneys were significantly lower than those of the normal kidneys. In renal artery stenosis kidneys, the ADC values in the cortex were significantly lower than those of the normal and the contralateral kidneys. In the cortex, ADC values were above 1.8 x 10(-3) mm2/sec in all 32 normal kidneys, ranging from 1.6 to 2.0 x 10(-3) mm2/sec in all 8 ARF kidneys, and below 1.5 x 10(-3) mm2/sec in 14 of 15 CRF kidneys. In the medulla, there was considerable overlap in the ADC values of the normal and diseased kidneys. There was a linear correlation between ADC value and sCr level in the cortex (r = 0.75) and a weak linear correlation in the medulla (r = 0.60). Our results show that diffusion-weighted MR imaging may be useful to identify renal dysfunction. PMID- 10373032 TI - Oxygen-enhanced magnetic resonance ventilation imaging of the human lung at 0.2 and 1.5 T. AB - Lung ventilation imaging using inhaled oxygen as a contrast medium was performed using both a 0.2 and a 1.5 T clinical magnetic resonance (MR) scanner in eight volunteers. Signal-to-noise-ratios (SNRs) of the ventilation images as well as T1 values of the lung acquired with inhalation of 100% oxygen and room air were calculated. The SNR was 9.7 +/- 3.0 on the 0.2 T MR system and 69.5 +/- 28.8 on the 1.5 T system (P < 0.001). The mean T1 value on the 0.2 T MR system with subjects breathing room air was 632 +/- 54 msec; with 100% oxygen, it was 586 +/- 41 msec (P < 0.01). At 1.5 T, the mean values were 904 +/- 99 msec and 790 +/- 114 msec, respectively (P < 0.0001). We conclude that MR oxygen-enhanced ventilation imaging of the lung is feasible with an open configured 0.2 T MR system. PMID- 10373033 TI - Evaluation of changes in intrarenal oxygenation in rats using multiple gradient recalled echo (mGRE) sequence. AB - Changes in intrarenal oxygenation in rats during pharmacological stimuli were evaluated with a multiple gradient-recalled echo (mGRE) sequence. With administration of the loop diuretic furosemide, oxygenation in the medulla improved; acetazolamide, a proximal tubular diuretic, produced no significant change. These results are consistent with our previous studies in humans and resemble earlier studies of medullary oxygenation using oxygen microelectrodes in anesthetized rats. The technique may be useful in the evaluation of therapeutic strategies in animal models of pathophysiological states such as acute renal failure. PMID- 10373034 TI - A novel object-independent "balanced" reference scan for echo-planar imaging. AB - Interleaved echo-planar imaging (EPI) is susceptible to significant ghosting artifacts, resulting primarily from system time delays that cause data matrix misregistration. Most EPI applications rely on "reference scans" to measure delays, and post-processing algorithms are used to correct these errors. Unfortunately, delay estimates made with most reference scan techniques are object dependent, since they are biased by magnetic field inhomogeneities and chemical shift. The current work describes the effects of field inhomogeneities and their influence on system time delay estimation. Subsequently, a new, object independent "balanced" reference method using two readout echo trains is proposed for time delay measurements. PMID- 10373035 TI - Subjects or participants? PMID- 10373036 TI - Strategic decisions of ice hockey coaches as a function of game location. AB - Two studies were performed to determine the influence of game location on the strategic decisions of ice hockey coaches. In study 1, coaches from the National (n = 23) and Ontario Hockey Leagues (n = 17) indicated the degree to which they had their teams forecheck assertively at home versus away. In study 2, video analysis of 62 National Hockey League games was used to verify the extent to which teams in this league use an assertive forechecking strategy at home versus away. In study 1, coaches reported that they implemented a more assertive forechecking style at home versus away (P < 0.001). The results of the video analysis in study 2 were consistent with the coaches' reports: teams used a more assertive forechecking style at home versus away (P < 0.03). The results are discussed in terms of their implications for the home advantage in the National Hockey League. PMID- 10373037 TI - Attitudes and exercise adherence: test of the Theories of Reasoned Action and Planned Behaviour. AB - Three studies of exercise adherence and attitudes are reported that tested the Theory of Reasoned Action and the Theory of Planned Behaviour. In a prospective study of adherence to a private fitness club, structural equation modelling path analysis showed that attitudinal and social normative components of the Theory of Reasoned Action accounted for 13.1% of the variance in adherence 4 months later, although only social norm significantly predicted intention. In a second study, the Theory of Planned Behaviour was used to predict both physical activity and sedentary behaviour. Path analyses showed that attitude and perceived control, but not social norm, predicted total physical activity. Physical activity was predicted from intentions and control over sedentary behaviour. Finally, an intervention study with previously sedentary adults showed that intentions to be active measured at the start and end of a 10-week intervention were associated with the planned behaviour variables. A multivariate analysis of variance revealed no significant multivariate effects for time on the planned behaviour variables measured before and after intervention. Qualitative data provided evidence that participants had a positive experience on the intervention programme and supported the role of social normative factors in the adherence process. PMID- 10373038 TI - Relationship between sport knowledge, sport performance and academic ability: empirical evidence from GCSE Physical Education. General Certificate of Secondary Education. AB - The literature concerning links between sport knowledge, sport performance and academic ability is reviewed and related to empirical evidence obtained from a GCSE examination in Physical Education, together with GCSE Mathematics and GCSE English grades. For most sports examined, there was a small but significant positive correlation between sport performance and GCSE Mathematics and English grades, confirming the findings of most previous research. Using a multilevel multivariate model, average sport performance, academic ability and sex were important explanatory variables for sport knowledge, yet only academic ability was an important explanatory variable for the concept of physical education knowledge. Ability in game sports, rather than athletics, were related to sport knowledge. Males scored higher for sport knowledge than females, after taking into account sport performance and academic ability. The effects of sport performance and academic ability on sport knowledge were stable across schools, but there was some evidence that the effect of sex varied across schools. These findings support theories of a role for sport knowledge in sport performance; that such a role should be greater in game sports; that academic ability is important for gaining such knowledge; and they highlight differences in sport knowledge between the sexes. PMID- 10373039 TI - On the presence and absence of behavioural traits in sport: an example from championship squash match-play. AB - Here we report two experiments that analysed sport (squash) competition as a non linear system that transits intermittently between different behavioural states. The first experiment involved a perceptual analysis of 60 rallies in which stable behaviour and unstable behaviour, delineated by behavioural transitions (i.e. shot perturbations), were reliably (kappa = 0.930) and validly (kappa = 0.844) identified by independent observers. In addition, experts were better than non experts at identifying the type of system behaviour (P < 0.01). These results provide for three alternative descriptions: (a) the system is multi-stable and transits between states via the mechanism of instability; (b) the system is bi stable and abruptly transits between two states, labelled stable and unstable; or (c) the system is mono-stable and displays variability, marked by transient instability, as a result of system perturbations. The second experiment analysed squash behaviour as expressed in the phase relation between the two players from time-motion analysis. The data, from four rallies, yielded evidence of a tight anti-phase coupling with transient phase shifts, or perturbations, that were quickly damped. These data suggest a mono-stable system with a single (anti phase) attractor onto which system fluctuations are occasionally written. However, these fluctuations failed to correspond with the short perturbations that were identified from perceptual analysis. Together, these results affirm the presence of transient behaviours in squash match-play, although the information that forms these perceptual judgements has yet to be identified. PMID- 10373041 TI - New insights into the effect of wind assistance on sprinting performance. AB - Here, a new mathematical model of sprinting is proposed. The prediction method rests on the construction of an energy balance incorporating a mathematical representation of each of the major terms in the balance. The term expressing the degradation of mechanical energy into thermal energy is formulated to express a dependence on wind speed. The dependence of the drag term on the change in mean body angle relative to the horizontal is taken into account. Whereas the effect of modifying the degradation term is shown to be significant, changes in body lean angle are shown to have little effect. Comparisons of the present predictions with a previous statistical analysis of 100-m track data show good agreement. Sensitivity analyses show which variables have the greatest influence on sprinting performance. PMID- 10373040 TI - A two-segment simulation model of long horse vaulting. AB - The optimum pre-flight characteristics of the Hecht and handspring somersault vaults were determined using a two-segment simulation model. The model consisted of an arm segment and a body segment connected by a frictionless pin joint, simulating the vault from the Reuther board take-off through to landing. During horse contact, shoulder torque was set to zero in the model. Five independent pre flight variables were varied over realistic ranges and an objective function was maximized to find the optimum pre-flight for each vault. The Hecht vault required a low trajectory of the mass centre during pre-flight, with a low vertical velocity of the mass centre and a low angular velocity of the body at horse contact. In contrast, the optimum handspring somersault required a high pre flight trajectory, with a high angular velocity of the body and a high vertical velocity at horse contact. Despite the simplicity of the model, the optimum pre flights were similar to those used in competitive performances. PMID- 10373042 TI - Methodological considerations in the determination of projected frontal area in cyclists. AB - Several studies have reported values for projected frontal area in cycling. Even when similar systems (i.e. riders and bicycles) have been measured, the results have diverged widely. It seems likely that this variability is due to methodological differences. The aims of the present study were to compare three methods of determining the frontal area in cyclists, and to determine the effects on the measured frontal area of variables which contribute to distortion and perspective in photographs. Theoretical models were developed to describe the expected effects of changing the relative position of the cyclist and the reference dimension, the position of the camera relative to the cyclist, and the focal length of the camera. Photographs were then taken of cyclists using different camera positions and settings, and analysed using three different methods: photographic weighing and manual and computerized planimetry. All three methods showed high precision and reliability, and yielded results that were substantially similar (mean values differed by <3.3%). Of possible sources of error, frontal deviation of the reference dimension had the most dramatic effect. Displacing the reference board forward by 0.4 m decreased the measured frontal area by 25%. As the distance between the camera and the cyclist increased, the frontal area decreased by about 5% for each metre. As the focal length of the camera became shorter, the frontal area became smaller, varying by >8% for focal lengths ranging from 28 to 70 mm. These results showed close agreement with the theoretical models, and can be explained in terms of the perspective and distortion effects which occur in photography. The results demonstrate the importance of standardization in measuring the frontal area of cyclists. PMID- 10373043 TI - Homebuilt aircraft crashes. AB - BACKGROUND: While the number of general aviation crashes has decreased over the 5 yr prior to 1993, the total number of homebuilt aircraft crashes has increased by nearly 25%. Research was undertaken to analyze these crashes and identify causal factors or unique problems associated with homebuilt aircraft. METHODS: Some 200 National Transportation Safety Board computer records and two-page descriptive briefs were analyzed for homebuilt aircraft crashes during 1993. Using descriptive epidemiology, variables were looked at in detail and comparisons were made with general aviation crashes during the-same year. RESULTS: Despite accounting for only 3% of all hours flown in general aviation certified aircraft for 1993, homebuilt aircraft accounted for 10% of the crashes and experienced a higher fatal crash rate. Crashes due to mechanical failure and crashes on takeoff and climb were more common in homebuilt aircraft as compared with general aviation. Other significant causal factors for homebuilt aircraft crashes included: minimal flight time in type specific aircraft, improper maintenance and improper design or assembly. CONCLUSIONS: Greater emphasis needs to be placed on educating homebuilt aircraft owners in the importance of following Federal Aviation Administration guidelines for certification and air worthiness testing. Understanding the aircraft's specifications and design limitations prior to the initial flight and properly maintaining the aircraft should also help to reverse the trend in the number of these crashes and subsequent lives lost. A system for assuring that all home-built aircraft are certified and more accurate reporting of flight hours are needed for accurate tracking of homebuilt aircraft crash rates. PMID- 10373044 TI - British Airways flightdeck mortality study, 1950-1992. AB - OBJECTIVE: To study the mortality and life expectancy of male British Airways flightdeck crew and to establish whether proportionate mortality excesses shown earlier for brain/CNS cancer, colon cancer and melanoma remained evident. METHODS: A Standardized Mortality Ratio study (SMR) using England and Wales as the comparison population was carried out for 6209 male pilots and 1153 male flight engineers employed for at least 1 yr between January 1, 1950 and December 31, 1992. Internal relative risk comparisons were made between shorthaul and longhaul operations defined broadly as flights within Europe and beyond Europe, respectively. RESULTS: The all-causes SMR for pilots of 61 (592 deaths) and 56 for flight engineers (127 deaths) confirmed the expected Healthy Worker Effect. In pilots apart from the known excess of deaths from aircraft accidents (SMR 14694), most of the comparisons showed significant deficits in mortality. The SMR's for brain/CNS cancer (143) and colon cancer (111) were no longer statistically significant. The SMR of 333 for melanoma was significantly raised in pilots but was not evident in flight engineers. Life expectancy for longhaul pilots and flight engineers was 4-5 yr better than England and Wales for ages 55 65 while the advantage for shorthaul pilots was reduced to between 2-3 yr. Cases of leukemia and aleukaemia in pilots were less than expected and less than the positive excess predicted from modeling based on radiation dose. CONCLUSION: The study confirms that flightdeck crew live longer than the England and Wales population and do not exhibit patterns of death that could be directly attributable to occupation. PMID- 10373046 TI - Echocardiographic evaluation of female centrifuge subjects for chronic changes in cardiac function. AB - BACKGROUND: High sustained G exposure as experienced in flying high performance aircraft can affect cardiac function. Numerous studies, mostly on male pilots, have evaluated the chronic effects of exposure to high G. To date, none of these studies has revealed significant positive findings in cardiac function as a result of long-term high G exposure. METHODS: A longitudinal study was conducted on six female centrifuge panel members who did not have a history of significant high +Gz exposure. Baseline echocardiographic studies were conducted prior to any +Gz exposure on the Dynamic Environment Simulator (DES) centrifuge. The echocardiograms were repeated after each panel member completed approximately 100 3-min high G (up to 9 G) exposures over the period of 7 mo. These follow-up echos were performed after all six subjects had been exposed to at least 6 h (cumulative) of sustained acceleration > 3 G. The women were protected with the COMBAT EDGE positive pressure breathing G protection ensemble. Each subject served as her own control. All studies were evaluated independently by a cardiologist who was blinded to the order in which the echos were performed. Although complete echocardiographic studies were performed, only the parameters identified as significant in prior studies were evaluated. RESULTS: No significant differences were found between the initial and follow-up echo parameters. CONCLUSIONS: We found no significant differences in cardiac function after at least 6 and up to 17 h (cumulative) of exposure to G > 3 in women. These subjects will be monitored during a longitudinal study throughout their centrifuge subject career. PMID- 10373045 TI - Muscle fatigue caused by repeated aerial combat maneuvering exercises. AB - BACKGROUND: Little is known about the development of in-flight muscular fatigue during repeated flights. HYPOTHESIS: This study was conducted to evaluate muscular fatigue in different upper body and neck muscles during repeated aerial combat maneuvering exercises. METHODS: Six pilots volunteered as test subjects. They performed one-to-one dog fight exercise three times (1 pilot, four times) in one day. During the flights, the pilots' electromyographic activity (EMG) was measured from the abdomen, back, neck and lateral neck. The mean muscular strain for each muscle was calculated. Before the first flight and after each flight, the maximal isometric strength of each muscle was measured. RESULTS: The results showed that maximal isometric strength between the first and last measurement decreased in the back, neck (p < 0.05) and lateral neck muscles. While the G stress remained the same, the muscular strain during exercises increased in every muscle, but was significant only in neck and lateral neck (p < 0.05-0.01). Due to these changes, the fatigue index in the neck and lateral neck muscles was 2.0 2.1, and 1.3-1.4 (1.0 = no fatigue) in the abdomen and back muscles. CONCLUSIONS: Repeated aerial combat maneuvering exercises caused fatigue in every muscle studied. The fatigue was greater in the neck area, which may increase the risk for neck injuries, and may reduce mission effectiveness. The fighter pilots' muscular strength and endurance in the neck area are subjected to very high demands, especially if exercises are repeated several times. The recovery of the neck muscles from fatigue after repetitive exercises should receive special attention. PMID- 10373047 TI - Neuroendocrine responses to psychological workload of instrument flying in student pilots. AB - BACKGROUND: Information processing and stress tolerance are necessary features for instrument flying (IFR), especially among student pilots. Psychological workload of IFR flight may lead to stress reactions such as neuroendocrine activity. METHODS: Neuroendocrine responses to an IFR flight with Vinka piston engined primary trainer were studied in 35 male volunteers who participated in the basic military flying course of the Finnish Air Force (FAF). The student pilots performed a 40-min IFR flight mission and a control session on land in randomized order between 11.00 h and 15.00 h. The IFR flight included 3 NDB approaches and was evaluated by flight instructors. Blood samples were collected 15 min before, 5 min and 60 min after the flight as well as control session, and. Plasma ACTH, beta3-endorphin (BE), cortisol, prolactin, adrenaline (A) and noradrenaline (NA) were measured. Psychological evaluations included psychomotor test (Wiener), Multi Coordination and Attention Test, ability tests and personality tests (CMPS and 16 PF). The overall psychological evaluation was made by an aviation psychologist. RESULTS: Plasma ACTH was significantly higher before and 5 min after the flight compared with control levels, but plasma BE increased significantly only before the flight. Plasma cortisol was significantly elevated before and 5 min after the flight. Plasma prolactin, NA and A increases were significant 5 min after the flight. High A levels after the flight correlated significantly with poor IFR flight performance as well as with poor psychomotor test results. CONCLUSIONS: The plasma prolactin and NA increases after the flight represented a direct type of stress reaction to the flight situation. The plasma BE response to IFR flight was an anticipatory stress reaction, but plasma ACTH, cortisol and A responses included both anticipatory and direct types of stress reactions. Psychological factors, flight performance and neuroendocrine responses to IFR flight appear to be associated with each other. Therefore, neuroendocrine reactions as a response to the psychological workload of military flying could be used for identifying stress tolerance in military pilots. PMID- 10373048 TI - Neuroendocrine responses and psychomotor test results in subjects participating in military pilot selection. AB - BACKGROUND: Military flying sets high demands on the mental performance and stress tolerance of pilots. Neuroendocrine responses could be a method for evaluating stress tolerance. METHODS: Psychological workload and neuroendocrine responses associated with the psychomotor Wiener's test were studied in 80 male volunteers. These personnel had applied for the basic military flying program of the Finnish Air Force (FAF). After the first blood sample at 0930 h, the subjects were randomly assigned to test (n = 40) and control groups (n = 40). The test group performed the psychomotor test, which lasted 10 min. The second blood sample was collected 1 min after the test. The control group was clinically examined and the blood sample was taken in the same way. RESULTS: A high plasma ACTH level before the psychomotor test predicted (r = 0.36, p = 0.02) a poor overall result in the psychomotor test. After the psychomotor test, plasma adrenocorticotropin (ACTH) and beta-endorphin levels were significantly higher than before the test. They were also higher than in the control group. Plasma cortisol and prolactin levels increased after the psychomotor test, but the increase was not statistically significant. High ACTH, cortisol and prolactin increments were specific to a high amount of delayed (over 2 s) psychomotor responses, as a marker of information overload. CONCLUSIONS: Elevated plasma ACTH, cortisol and prolactin levels, after the psychomotor test, were associated with a high amount of the delayed responses. This indicates that high neuroendocrine responses were connected with problems in stress tolerance during information processing. High neuroendocrine reactions under information load could, therefore, be used for identification of lowered stress tolerance. PMID- 10373049 TI - Moderate sodium restriction does not alter lower body negative pressure tolerance. AB - HYPOTHESIS: Space travel with exposure to microgravity leads to a significant reduction in orthostatic tolerance on return to Earth, for which countermeasures are only partially successful. The purpose of this study was to examine the effect of moderate dietary sodium restriction on tolerance to LBNP. METHODS: Eight healthy men, age 25.1+/-1.3 yr, volunteered for the study. Subjects were exposed to presyncopal LBNP after consuming their "typical" diet (C) for 5 d and after consuming a sodium restricted (SR) diet for 5 d. Diet sequence was randomized and adherence was verified by 24-h urine collection on the 4th and 5th days of each diet. RESULTS: All subjects reached presyncope during the LBNP, regardless of diet. Urinary sodium excretion was 3390+/-950 mg on the C diet and 1174+/-560 mg on the SR diet. Urinary potassium was not different between the diets. Cumulative stress index scores were 655+/-460 (mm Hg x min) on the C diet and 639+/-388 (mm Hg x min) during SR. Cardiac volumes, BP and total peripheral resistance were not different at any stage of the LBNP between the diets, nor were catecholamines. Plasma renin activity, determined by radioimmunoassay, was significantly higher during SR at rest, and during all stages of LBNP in comparison with the control diet. CONCLUSION: Moderate dietary sodium restriction is not detrimental to orthostatic tolerance. PMID- 10373051 TI - Fibroblast response to rapid decompression and hyperbaric oxygenation. AB - INTRODUCTION: The cellular basis for symptoms associated with rapid decompression and the use of hyperbaric oxygenation treatment (HBO) is not established. METHODS: Image analysis, sulforhodamine B assay and bromodeoxyuridine (BrDU) incorporation were used to identify cellular changes associated with rapid decompression (RD) or hyperbaric oxygenation (HBO). Human fibroblasts were exposed to RD or HBO and compared with untreated cells. Immediately following treatment, the fraction of cells synthesizing DNA was measured by detection of cells incorporating the BrDU. At 1 d and 3 d following the treatments, total cell protein adherent to the bottom of wells was measured using a sulforhodamine B assay. Cell density was observed with light microscopy and quantified with image analysis. RESULTS: RD increased total protein significantly, (p < 0.05) relative to control, while HBO had less effect. The fraction of cells synthesizing DNA was increased by HBO and reduced by RD relative to control (p < 0.05). Image analysis showed that cell density at day 1 was: control>HBO>RD; and at day 3: RD>HBO>control, indicating an increase in proliferation induced by the treatments. CONCLUSION: This data shows that HBO and RD increase the proliferation of fibroblasts for 24 h following treatment. HBO increased DNA replication. While there was a decrease in DNA replication following RD, protein synthesis was enhanced. PMID- 10373050 TI - Thermal regulation in the heat during exercise after caffeine and ephedrine ingestion. AB - BACKGROUND: Ingesting a combination of caffeine and ephedrine (C+E) has been shown to raise metabolic heat production and body temperature. This side effect of C+E ingestion may be positive during a cold stress scenario, however, during heat stress it could prove to be detrimental. Thus, the purpose of this study was to clarify the effect of C+E ingestion on body temperature regulation during moderate exercise in a hot dry environment. METHODS: Ten, healthy, non heat acclimated, males exercised at 50% VO2peak in a 40 degrees C and 30% RH environment until rectal temperature reached 39.3 degrees C; heart rate (HR) remained at 95% of peak value or greater for 3 min, dizziness or nausea precluded further exercise, or 3 h had elapsed. They did this four times at weekly intervals: familiarization (Fam), control (Cont), placebo, and C+E (5 mg . kg(-1) caffeine + 1 mg . kg(-1) ephedrine) trials. The Fam and Cont treatments were done first and sequentially while the placebo and C+E treatments were balanced and double-blind. Tolerance times, mean skin temperature (Tsk), rectal temperature (Tre), Vo2, Vco2, VE, sweat rate (SR), HR, and sensation of thermal comfort were measured. RESULTS: Tolerance times (mean+/-SD in minutes) were similar for the placebo (120.0+/-28.4) and C+E (121.3+/-33.9) trials and both times were significantly longer than Cont (106.6+/-24.0) trial. C+E did not affect Tsk, initial TrC, delta Tre, SR or the sensation of thermal comfort. VO2 and VF, were significantly increased by C+E. HR was elevated by C+E compared with the other trials, but only during the initial 20 min of exercise. CONCLUSION: Although the metabolic rate was slightly increased with C+E treatment, it was sufficiently offset by increased heat loss mechanisms so that internal body temperature was not increased during moderate exercise in a hot, dry environment. PMID- 10373052 TI - Hyperbaric chamber-related decompression illness in a patient with asymptomatic pulmonary sarcoidosis. AB - An asymptomatic 46-yr-old male sustained an acute neurologic insult, appearing during the decompression phase of a 50-m dry hyperbaric chamber dive. The right hemisyndrome was most probably related to diving, since symptoms responded rapidly to the early commenced recompression therapy. Further diagnostics revealed a previously unknown pulmonary sarcoidosis with bilateral pulmonary opacities and pleural adhesions that might have predisposed to arterial gas embolism secondary to pulmonary barotrauma. This case may illustrate a potential risk of decompression illness even during dry chamber dives in patients suffering from asymptomatic pleuro-parenchymal pulmonary disease. The value of chest X-ray in the medical assessment of fitness to dive is therefore emphasized. PMID- 10373053 TI - Reduce risk of inducing spatial disorientation using physiologically compatible ground lighting. AB - Spatial disorientation that occurs while landing aircraft during night operations may result in accidents and fatalities which are often classified as secondary to "pilot error." It has now been determined that the use of "expedient" lights, which include flares, flashlights, automobile headlights, etc., can induce spatial disorientation in pilots. The element that contributes to induction of spatial disorientation is the "point source of light" provided by these lights. Impingement on the retina of concentrated photon emissions, as supplied by incandescent (filament) lamps, flares, etc., produces an "after image," such as occurs when one briefly looks into the sun. The "after image" is caused by the time lag required for reconstitution of the neurohumoral transmitter substances in the retinal rods and cones. Pilots who develop "after images" during the final stage of landing a helicopter at night are predisposed to experiencing spatial disorientation, leading to aircraft mishaps. In contrast to flares or incandescent light sources, cold cathode lamps lack a "point source" of light emission, do not create an "after image," and are ideal to use in night landing operations. Cold cathode lights operating in the range of 512 nm (blue-green) are thought to be the most physiologically efficient color to use for night landing operations. Light sources in this nanometer range provide maximum visibility and safety for the pilot during landing operations under all environmental conditions. PMID- 10373054 TI - Cognitive performance over 7 days in a distressed submarine. AB - BACKGROUND: This article contains the results from a psychological assessment of submariners undergoing a survival trial simulating conditions in a disabled submarine. The aim was to determine whether the environmental conditions and rations in a submarine escape compartment had any detrimental effects on cognitive performance. METHODS: The study was conducted in an environmental chamber in which the temperature fell from 22 degrees C to 4.4 degrees C over 2 d and then remained at 4.4 C for 5 d. Of the 11 subjects who were given daily rations of 100 g of barley sugar and 568 ml of water (none on day 1), only 4 remained for the full trial duration. Subjects were administered 2 psychometric tests (choice reaction time and short term memory) on hand held computers. RESULTS: Results showed that subjects did not exhibit any significant performance decrements during the experimental phase of the trial. However anecdotal evidence from observations and subjects' self reports suggest that sustained performance was impaired. PMID- 10373055 TI - Specialty competencies for residents in aerospace medicine. AB - BACKGROUND: The American College of Preventive Medicine (ACPM), with sponsorship from the Health Resources Administration (HRSA), has published core competencies that are common to all preventive medicine residencies-aerospace medicine (ASM), occupational medicine (OM), and general preventive medicine/public health (GPM/PH). Further development of specialty area competencies for ASM residents was addressed by a working group comprised of representatives from each of the four ASM residency programs. METHODS: Representatives from the U.S. Air Force School of Aerospace Medicine, Wright State University, University of Texas Medical Branch-Galveston, and the Naval Operational Medicine Institute convened to develop a set of broad competency statements for ASM residents that would encompass the breadth of ASM residency training as it is currently provided in the U.S. RESULTS: A listing of six ASM resident competencies, with supporting skill sets, are presented. In combination with the ACPM core competencies, the ASM resident competencies represent a refocusing of educational objectives on skills attainment. CONCLUSIONS: The ASM resident competencies identify the capabilities of graduating ASM residents as distinct from OM and GPM/PH residents. At the same time, they are broad enough to permit specific areas of emphasis (e.g., military, civil, or space) to be pursued within the various ASM residencies. This represents the first successful attempt to draft a consolidated statement of educational objectives that has universal acceptance and applicability across all U.S. aerospace medicine residencies. PMID- 10373056 TI - Samuel Franklin Cody: aviation and aeromedical evacuation pioneer. AB - In the summer of 1913, Woodrow Wilson had just become the 28th President of the United States and King George V was on the throne of the United Kingdom. It was nearly 10 yr since the historical flight at Kitty Hawk and 3 yr since Bleriot had first flown the English Channel. At Farnborough, "Colonel" Samuel F. Cody, originally a horseman, hunter, crack shot, showman and theatrical impresario from the USA, was preparing a new floatplane for a round Britain flying race. One of the features of the floatplane, a biplane with a four bladed pusher propeller, was that it had already demonstrated its ability to carry passengers. Cody calculated that it would allow him to carry five passengers for a 4-h flight and may even have medical uses. He arranged a demonstration of its potential as an air ambulance at Farnborough. This paper describes, with the use of photographs of the event, the airplane, the demonstration and the reasons why it would be left to others to carry out the first, real aeromedical evacuation 4 yr later. PMID- 10373058 TI - Magnesium and acute mountain sickness. PMID- 10373060 TI - The gyroplane. PMID- 10373061 TI - Thiotepa, busulfan and cyclophosphamide as a preparative regimen for allogeneic transplantation for advanced chronic myelogenous leukemia. AB - Thirty-six adults with chronic myelogenous leukemia (CML) in second or greater chronic phase, accelerated phase, or blast crisis underwent marrow or blood stem cell transplantation from an HLA-matched sibling using high-dose thiotepa, busulfan and cyclophosphamide (TBC) as the preparative regimen. All evaluable patients engrafted and had complete donor chimerism. One patient failed to clear meningeal leukemia, and one patient had one of 30 metaphases positive for the Philadelphia chromosome at 2 months post transplant. The remainder of the patients studied had eradication of CML documented by cytogenetics and/or Southern blot for BCR gene rearrangement, and 13 of 15 patients studied became negative for the BCR gene rearrangement by polymerase chain reaction. Three-year relapse rate is 42% (95% CI, 19-64%). The relapse rate was significantly lower for patients transplanted without blast crisis (9% vs 100%, P < 0.001). Eight (22%, 95% CI, 10-39%) patients had severe or fatal veno-occlusive disease (VOD). Elevated liver enzymes within 1 month prior to transplantation and transplantation using marrow were significantly associated with the occurrence of VOD. Three-year survival is 28% (95% CI, 13-43%). Survival was significantly higher for patients transplanted without blast crisis (45% vs 0%, P = 0.01). TBC is an effective preparative regimen for CML in accelerated phase but not refractory blast crisis, and it should be used with caution in patients with prior hepatopathy who have an increased risk of severe VOD. PMID- 10373062 TI - Influence of recombinant human granulocyte colony-stimulating factor (filgrastim) on hematopoietic recovery and outcome following allogeneic bone marrow transplantation (BMT) from volunteer unrelated donors. AB - Effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF, filgrastim) on hematopoietic recovery and clinical outcome in patients undergoing allogeneic bone marrow transplantation (BMT) from volunteer unrelated donors (VUD) were analyzed retrospectively. Additionally, the influence of baseline patient and transplant characteristics on hematopoietic recovery was evaluated. From January 1994 to March 1996, 47 consecutive adult patients received VUD-BMT. GVHD prophylaxis was cyclosporin A/short course methotrexate/prednisolone, and in four patients additional ATG. Post-transplantation, cohorts of patients received rhG-CSF (5 microg/kg/day) (n = 22) or no rhG-CSF (n = 25) in a non-randomized manner. The patient groups with and without rhG-CSF were rather comparable with respect to baseline patient and transplant characteristics. Median time to neutrophil counts (ANC) >500/microl was 14 days with rhG-CSF vs 16 days without rhG-CSF (P = 0.048), to ANC >1000/microl was 15 vs 18 days (P = 0.084). Neutrophil recovery was accelerated in patients receiving more than the median MNC dose of 2.54 x 10(8)/kg with a median time to ANC >1000/microl of 13 days vs 19 days (P = 0.017). RhG-CSF did not influence platelet recovery and incidence of infectious complications. Incidence of acute GVHD II-IV was 50% with rhG-CSF and 28% without rhG-CSF (P = 0.144), but death before acute GVHD II-IV occurred in 9% of patients with and 20% of patients without rhG-CSF. The median follow-up time was 38 and 36 months in patients with and without rhG-CSF, respectively. Survival at 2 years post-transplant was 39% (95% confidence interval (18%, 60%)) in patients with rhG-CSF and 24% (95% confidence interval (7%, 41%)) in patients without rhG-CSF. Administration of rhG-CSF after VUD-BMT may lead to more rapid neutrophil recovery, but did not influence the incidence of infectious complications. Patients receiving rhG-CSF showed a slightly higher incidence of acute GVHD II-IV. Higher numbers of MNC in the marrow graft accelerated hematopoietic engraftment. PMID- 10373063 TI - A prospective study of G-CSF effects on hemostasis in allogeneic blood stem cell donors. AB - Granulocyte colony-stimulating factor (G-CSF) is used in healthy donors of peripheral blood stem cells (PBSC) for allogeneic transplantation. However, some data have recently suggested that G-CSF may induce a hypercoagulable state, prompting us to study prospectively 22 PBSC donors before and after G-CSF 5 microg/kg twice daily. We sought evidence for changes in the following parameters: platelet count, von Willebrand factor antigen (vWF:Ag) and activity (vWF activity), beta-thromboglobulin (beta-TG), platelet factor 4 (PF-4), platelet activation markers (GMP-140 and PAC-1), activated partial thromboplastin time (aPTT), prothrombin time (PT), coagulant factor VIII (FVIII:C), thrombin antithrombin complex (TAT), prothrombin fragment 1+2 (F1+2), thrombomodulin (TM) and tissue plasminogen activator antigen (tPA:Ag) prior to G-CSF and immediately before leukapheresis. ADP-induced platelet aggregation studies were also performed. G-CSF administration produced only mild discomfort. We found a significant increase in vWF:Ag (from 0.99 +/- 0.32 U/ml to 1.83 +/- 0.69 U/ml; P < 0.001), in vWF activity (from 1.04 +/- 0.34 U/ml to 1.78 +/- 0.50 U/ml; P < 0.001) and in FVIII:C (from 1.12 +/- 0.37 U/ml to 1.73 +/- 0.57 U/ml; P < 0.001) after G-CSF. Of note, four donors with low baseline vWF had a two- to three-fold increase after receiving G-CSF. G-CSF had no impact on the platelet count, beta TG, PF-4, GMP-140 or PAC-1. The final% of platelet aggregation decreased from 73 +/- 22% to 37 +/- 26% after G-CSF (P < 0.001). We found a significant decrease in aPTT after G-CSF (29.9 +/- 3.1 s to 28.3 +/- 3.3 s; P = 0.004), but the PT was unaffected. In addition, we also observed a significant increase in TAT, F1+2 and TM, but not in tPA:Ag. Our data suggest that G-CSF may possibly induce a hypercoagulable state by increasing levels of FVIII:C and thrombin generation. In contrast to this information, we found reduced platelet aggregation after G-CSF administration. The clinical implications of these findings remain unclear and larger studies are definitely required. PMID- 10373064 TI - Diagnosis of secondary myelodysplastic syndromes (MDS) following autologous transplantation should not be based only on morphological criteria used for diagnosis of de novo MDS. AB - Secondary myelodysplastic syndromes (MDS) are increasingly being reported after autologous transplantation. Transient dysplastic changes have also been observed after this type of treatment. However, to the best of our knowledge no systematic morphological analysis has been performed to determine the influence of stem cell transplantation on bone marrow morphology. In 53 patients undergoing autologous transplantation, we evaluated the bone marrow, before and 6 and 12 months after the transplant, in order to analyze the appearance of dyshemopoietic changes, assessed according to a pre-established score. We also studied 25 bone marrow samples obtained at the time of diagnosis, prior to treatment, but we did not find morphological atypia. Six months after transplant, cellularity and thrombopoiesis had decreased in 38% and 49% of patients respectively, although 1 year after the process they were normal in most cases. Myelodysplasia was already present in bone marrow before transplantation and continued to be in evidence for a long time afterwards. This suggests that chemotherapy and radiotherapy used prior to transplantation are responsible for dysplastic changes. The myeloid line was the most affected with 100% of patients showing dysgranulopoiesis 1 year after autografting. Cytopenias were observed in 51% and 44% of patients 6 and 12 months after transplantation. Moreover, concomitant presence of cytopenia and myelodysplasia was observed in 37.7% of patients at 6 months after transplantation and 25% at 12 months, and therefore they could be diagnosed with MDS. These data contrast with the incidence of secondary MDS reported in earlier publications. According to these findings, the value of the French-American British Co-operative Group criteria for the diagnosis of MDS following autologous transplantation is questionable. Moreover, since dyshemopoietic features are almost always present after autologous transplant, morphological criteria are not useful for early recognition of patients with secondary MDS after transplantation. PMID- 10373065 TI - Melphalan 220 mg/m2 followed by peripheral blood stem cell transplantation in 27 patients with advanced multiple myeloma. AB - Twenty-seven patients with advanced multiple myeloma received high-dose therapy with 220 mg/m2 i.v. melphalan (HDM220) followed by autologous stem cell transplantation. At the time of HDM220, nine patients had primary refractory disease and 18 were in relapse after having responded to prior high-dose therapy. No toxic deaths were observed. The major adverse side-effect was grade 4 mucositis in 63% of patients. Two patients experienced reversible paroxysmal atrial fibrillation after HDM220. For the whole group of patients, the actuarial 3-year overall survival (OS) and event-free survival (EFS) are 36.1 and 16.9%, respectively. The probability of OS and EFS was significantly lower in patients treated for refractory relapse (22.9 and 0% at 2 years, respectively) as compared to primary refractory patients (66.7 and 64.3% at 2 years, respectively) or patients treated for chemosensitive relapse (42.9% at 2 years) (P = 0.0001). Low beta2-microglobulin and CRP levels at the time of HDM220 were associated with a better OS and EFS. Our data suggest that HDM220 followed by ASCT should be considered in patients with primary refractory disease or chemosensitive disease relapsing after prior intensive therapy. PMID- 10373066 TI - Pilocarpine hydrochloride relieves xerostomia in chronic graft-versus-host disease: a sialometrical study. AB - Bone marrow transplantation is considered to be the treatment of choice for various hematological and solid malignancies, as well as for bone marrow failure syndromes and some genetic diseases. Unfortunately, a great number of patients who receive allogeneic BMT suffer from graft-versus-host disease (GVHD) following the procedure. Xerostomia is considered to be one of the most annoying complications of chronic GVHD (cGVHD), and the rapidly growing number of BMT patients with prolonged survival renders GVHD-related xerostomia a major clinical problem. As pilocarpine hydrochloride has been shown to relieve xerostomia in other disease categories, we administered pilocarpine hydrochloride 30 mg/day to six cGVHD patients and measured their whole saliva, parotid and submandibular sublingual flow rates in both resting and stimulated conditions. Mean values of flow rates of whole saliva in resting conditions at 2 weeks, 2 months and 6 months following administration of pilocarpine hydrochloride 30 mg/day were 0.71 +/- 0.12 ml/min, 0.59 +/- 0.07 ml/min and 0.56 +/- 0.11 ml/min, respectively. In stimulated conditions, mean values were 1.7 +/- 0.3 ml/min, 1.0 +/- 0.17 ml/min and 0.94 +/- 0.21 ml/min, respectively. The mean values of whole saliva flow rates under both conditions represented an increase of 224-284% and 134-247%, respectively (P < 0.01). The pattern and magnitude of parotid and submandibular sublingual flow rate increases following pilocarpine hydrochloride administration were similar. Patients were followed for 6 months and demonstrated increased levels of secretion, with some reduction after the initial peak values. The medication was discontinued at 2 months and reinstated after 2 weeks in three patients. This resulted in rapid flow rate reduction followed by another profound increase. Contrary to the sialometrical analysis, the subjective scoring showed no fluctuations during the study period. We discuss these results in the context of the clinical experience of xerostomic patients in whom even a minute increase in secretion may be significant. Our results demonstrate that objective and subjective relief from xerostomia in cGVHD patients can be achieved with the routine oral administration of pilocarpine hydrochloride. PMID- 10373067 TI - Expression of T cell activation antigen CD134 (OX40) has no predictive value for the occurrence or response to therapy of acute graft-versus-host disease in partial T cell-depleted bone marrow transplantation. AB - CD134 (OX40) is a member of the tumor necrosis factor family which is expressed by activated T lymphocytes. CD134 expression on T cells was monitored during the first 35 days post-transplant in 14 patients, receiving either an HLA-identical sibling bone marrow transplant (BMT), a matched unrelated transplant (MUD-BMT) or an autologous peripheral blood progenitor cell transplant (PBPCT). The sibling and unrelated grafts were partially depleted of T cells. CD134 expression on CD4+ T cells peaked between 7 and 14 days after BMT, with a mean peak value of 45% of CD4+ cells (range 26-70%) over all three patient groups. The observed pattern of CD4+ CD134+ expression, an increase during the first 2 weeks post-BMT followed by a gradual decline towards values of 15-40%, was similar in all groups. No difference in the kinetics of CD134 expression by CD4+ T cells was observed between the patients that did or did not develop graft-versus-host disease (GVHD), nor did the clinical effect of any treatment given for GVHD correlate with alterations in CD134 expression by CD4+ T cells. Absolute CD4+,CD134+ T cell numbers showed a more rapid increment after autologous PBPCT than after sibling or MUD transplants. We conclude that expression of CD134+ by CD4+ T lymphocytes cannot serve as a surrogate marker for allo-reactivity. CD134+ expression may reflect lymphocyte regeneration, rather than alloreactivity. PMID- 10373068 TI - Analysis of circulating tumor cells in patients with multiple myeloma during the course of high-dose therapy with peripheral blood stem cell transplantation. AB - In multiple myeloma (MM) circulating CD19+ cells have been considered as myeloma precursors. As these cells are also possibly a reservoir of treatment resistant disease evaluation of the CD19+ cells during the course of high-dose therapy has to be a major concern. We determined the number of tumor cells in the CD19+ as well as CD19- fractions of PB of eight patients with disease sensitive to VA[I]D chemotherapy, of 10 patients who achieved partial or complete remission post-high dose therapy (HDT) with peripheral blood stem cell transplantation (PBSCT) and of a further seven patients with disease progression post-transplantation. CD19+ cell fractions were obtained by preparative sequential magnetic and fluorescence activated cell sorting with a median purity of 97.1%. In addition, PB samples of seven patients post-transplantation were sorted for CD20+ cells (median purity, 98.7%). The number of tumor cells in the CD19+, the CD19- and the CD20+ fractions were determined using a quantitative CDR3 PCR assay. The number of CD19+ tumor cells in patients in remission post-HDT was similar to those of the patients post VA[I]D (median, 1.05 vs 0.92 CD19+ tumor cells/ml PB, P = 0.72) providing evidence for the persistence of this tumor cell fraction during the course of HDT. This was in contrast to the CD19- compartment, in which the number of tumor cells was significantly reduced in those patients in remission post transplantation (median, 53 vs 0 CD19- tumor cells/ml PB; P = 0.006). In patients with progressive disease the number of tumor cells in both cell fractions was significantly higher (CD19+: median, 1.05 vs 21 tumor cells/ml PB, P = 0.05; CD19 : 0 vs 63 tumor cells/ml PB, P = 0.008). While the absolute number of CD19+ cells was reduced in the group of patients after VA[I]D treatment, a polyclonal CD19+ reconstitution had occurred in patients responding to HDT. The tumor cell content in the CD19+ fractions could be confirmed by the results obtained analyzing the CD20+ cell fractions. In conclusion, these results indicate that disease progression after PBSCT in MM is accompanied by an expansion of tumor cells in both the CD19+ and CD19- fractions. Similar numbers of CD19+ clonotypic cells post-HDT suggest that these cells persist and thus, contribute to disease dissemination and relapse. PMID- 10373069 TI - Long-lasting decrease of marrow and circulating long-term culture initiating cells after allogeneic bone marrow transplant. AB - We investigated bone marrow (BM) and circulating (PB) hematopoietic progenitor cells in 37 normal donors and in 25 patients 1 to 8 years after successful allogeneic bone marrow transplant. At the time of testing, transplanted patients had normal blood counts and bone marrow cellularity. By flow cytometry, BM CD34+ cells were found to be three- to four-fold decreased in transplanted patients compared to normal donors, while the number of PB CD34+ cells was the same as in normal donors. Using a methylcellulose colony assay, primary BM colony-forming cells (CFU-GM) were decreased 2.1-fold, whereas PB CFU-GM were only marginally decreased. In a long-term culture initiating cell (LTC-IC) assay, an eight-fold decrease of early progenitor cells was observed in the marrow of transplanted patients compared to normal donors, and a five-fold decrease was documented in peripheral blood. We found that the BM LTC-IC cell number correlated with concurrently determined BM CD34+ cells and committed progenitor cell number (measured as CFU-GM) and with PB LTC-IC number, but not with PB CFU-GM and CD34+ cells. We conclude that marrow and circulating early stem cell compartments, as measured by the LTC-IC assay, are greatly and permanently depressed following bone marrow transplant. The correlation between BM and PB LTC-IC indicates that the enumeration of circulating LTC-IC can be used as a measure of the stem cell compartment in the bone marrow after transplant. It seems that the deficiency of the most immature progenitor cells persists forever after successful bone marrow transplant; this means that a complete hematopoietic reconstitution can be sustained by a reduced stem cell pool. PMID- 10373070 TI - Incidence and outcome of Clostridium difficile infection following autologous peripheral blood stem cell transplantation. AB - A retrospective evaluation of 200 consecutive recipients of autologous peripheral blood stem cell transplantation (PBSCT) was conducted to ascertain the incidence and outcome of infection with Clostridium difficile. The diagnosis was confirmed in 14 patients with diarrhea (15 episodes) at a median of 33 days after stem cell infusion. Five patients were neutropenic at the time of diagnosis. Every individual had adverse known risk factors such as recent or current use of antibiotic, corticosteroid and antiviral therapy, recent administration of myeloablative chemotherapy and numerous, prolonged periods of hospitalization. Diarrhea, frequently hemorrhagic, was the most common presenting feature along with fever, abdominal cramps and abdominal distention. Diagnosis was established by the stool-cytotoxin test. Response to standard treatment with oral vancomycin or metronidazole was prompt despite the presence of several adverse prognostic features in these patients. There was only one instance of relapse which was also treated successfully. Several transplant-related variables such as age, sex, underlying malignancy, myelo-ablative regimen, duration of neutropenia, and prophylactic use of oral ampicillin underwent statistical analysis but failed to be predictive of C. difficile infection in such a setting. Finally, C. difficile is not uncommon after autologous PBSCT and must be included in the differential diagnosis in any such patient with diarrhea. PMID- 10373071 TI - Nutritional status and growth after bone marrow transplantation (BMT) during childhood: EBMT Late-Effects Working Party retrospective data. European Group for Blood and Marrow Transplantation. AB - The European Group for Blood and Marrow Transplantation (EBMT) Late-Effects Working Party collected data on patients who survived more than 5 years after BMT. Height at transplant and at the latest follow-up examination were evaluated in 79/258 subjects who were below the age of 15 at BMT. A significant decrease in height-standard deviation score (SDS) was observed in leukemic children conditioned with total body irradiation (TBI) and in those who received both cranial irradiation and TBI. The majority of these patients, however, received single-dose TBI (28/41). A significant decrease in height-SDS was also seen in children who received thoraco-abdominal irradiation suggesting that the deleterious effect of irradiation on growth after BMT is not entirely due to injury to cranial neuroendocrine structures, but also probably due to damage to bone epiphyses, thyroid and gonads. A non-significant decrease in height was observed in children transplanted using chemotherapy alone. Nutritional status, expressed as body-mass index (BMI), was found unchanged in the adult group (n = 158). A significant increase in BMI was observed in the younger patients (n = 88), which parallels the normal increase in BMI observed during childhood. This suggests that on long-term analysis, a good nutritional status is maintained in patients undergoing BMT at any age. PMID- 10373072 TI - Organ toxicity and quality of life after allogeneic bone marrow transplantation in pediatric patients: a single centre retrospective analysis. AB - One hundred and fifty five pediatric patients underwent allogeneic bone marrow transplantation between 1980 and 1996 in the St Anna Children's Hospital in Vienna with an overall survival of 52.3% (81 patients). Seventy-three patients with a minimum observation time of 1 year (1-13 years, median: 4.6) were analyzed retrospectively for chronic GVHD, organ toxicity (WHO score), growth and pubertal development. Chronic GVHD was diagnosed in 20 patients (27.3%), being extensive in 17 cases. Maximum organ toxicity was WHO III in two patients (3%) and WHO II in 11 patients (15%) 1 year after BMT and WHO III in one patient (2%) and WHO II in five patients (11%) 3 years after BMT. Impaired growth and pubertal development were detected in more than 30% 3 years after BMT. As all patients presented with a Karnofsky or Lansky score of more than 80%, they were asked to complete a questionnaire comprising 12 questions concerning physical state of health and psychosocial state of health. Restricted contacts were classified as imposing a severe handicap by six patients (8%), restriction in mobility and 'normal life activities' by three patients (4%) and two patients classified themselves as severely physically handicapped. Most patients (75%) reported no physical or psychical impairment. PMID- 10373073 TI - Rapid quantification of mixed chimerism using multiplex amplification of short tandem repeat markers and fluorescence detection. AB - Monitoring the engraftment of donor cells after allogeneic blood stem cell transplantation (BSCT) may be important for the early diagnosis of graft failure or relapse of disease. Several techniques have been reported for this purpose. PCR-based assays analyzing polymorphic short tandem repeat (STR) markers are attractive because they are sensitive and can be performed rapidly. The intent of the present study was to test a novel approach for the quantification of mixed chimerism using a commercial multiplex STR assay with fluorescence-based detection for forensic purposes. The feasibility of this assay and the accuracy of quantitative results was tested using serial cell mixtures of unrelated individuals. Sample preparation was optimized to obtain information from minute amounts of starting material, eg from patients with aplasia or from sorted cell populations. Using the STR-PCR, discrimination between donor and recipient was possible in all patients analyzed (n = 25). Cell dilution experiments showed a linear correlation between the cell numbers added and the proportions found, with the limit of detection for a minor cell population being 5%. Comparison of values obtained with standard FISH analysis in patients transplanted from sex-mismatched donors showed an excellent correlation with the STR-PCR results. Taken together, this procedure allows the rapid, versatile and accurate quantification of mixed chimerism, even with minuscule numbers of cells. PMID- 10373074 TI - Counterflow centrifugation allows addition of appropriate numbers of T cells to allogeneic marrow and blood stem cell grafts to prevent severe GVHD without substantial loss of mature and immature progenitor cells. AB - Using counterflow centrifugation elutriation (CCE) lymphocytes can be separated from CD34+ populations based on size. Immature progenitors tend to be smaller than mature cells suggesting that CCE introduces loss of stem cells. We compared the separation of 12 PBSC with 16 BM transplants. Cells were separated in 12 fractions (3000-2200 r.p.m.) and the rotor off (RO) fraction. Separation patterns of BM and PBSC were comparable. B cells were collected in the high speed fractions followed by T and NK cells. In contrast, progenitor cells were collected in lower speed fractions. By adding successively T cell-depleted fractions to the RO fraction a BM transplant could be composed containing 0.7 x 10(6) T cells/kg and 90%, 89% and 68% recovery of CD34+, CFU-GM and BFU-E. PBSC were separated in four CCE runs inducing higher numbers of T cells in the graft (4.4 x 10(6)/kg) and 54% CD34+, 46% CFU-GM and 37% BFU-E recovery. Time of engraftment was not delayed and no graft failure was observed. The higher number of T cells was not associated with higher incidence of GVHD. Acute GVHD > or = grade III occurred in 0 of 16 BM and two of 12 PBSC recipients; extensive chronic GVHD was observed in four of 15 and three of nine recipients, respectively. To study immature cells in the graft, CD34 subpopulations and cells with long-term repopulating ability, determined using cobble-stone area formation (CAFC assay), were evaluated in each fraction. The separation patterns in BM and PBSC were comparable. Cells with mature and immature phenotype were enriched in lower speed fractions (mean recovery of 74% CD34+/CD13-/DR-). The CAFC week 2, 4 and 6 were also enriched in these fractions. These data show that the used CCE procedure is a reliable method to deplete T cells from stem cell transplants without substantial loss of immature and mature progenitors. PMID- 10373075 TI - Selective removal of alloreactive cells from haematopoietic stem cell grafts: graft engineering for GVHD prophylaxis. AB - One of the main goals in allogeneic bone marrow transplantation is the abrogation of graft-versus-host disease with the preservation of antileukaemia and antiviral activity. We have established a novel system for the selective removal of alloreactive lymphocytes from donor grafts while retaining an effective allogeneic response to third-party stimulator cells. Initial feasibility studies were done with unrelated HLA-mismatched pairs and then extended into the matched setting. Mononuclear cells from HLA-matched donors were cocultured with irradiated recipient cells prestimulated with cytokines (gamma-IFN and TNF-alpha) in a modified mixed lymphocyte culture (MLC). Alloreactive donor lymphocytes were identified by expression of CD69, an early activation marker and selectively removed by paramagnetic bead sorting. The remaining 'non-alloreactive' lymphocytes were tested in proliferative assays against the original matched recipient and to a third-party donor. A mean depletion of proliferative capacity to 11.5 +/- 9.9% of the original matched recipient response was achieved while the residual third-party response was largely preserved at 77.8 +/- 20.9% which should translate into improved immune reconstitution and preservation of antiviral activity. The non-alloreactive lymphocytes could also possess functional antileukaemia activity. Moreover, the alloreactive cells are easily recoverable in this selective T cell depletion strategy for cryopreservation and ready for immediate access as therapeutic donor lymphocyte infusions in cases of frank relapse post transplant. PMID- 10373076 TI - Successful bone marrow transplantation in a child with X-linked hyper-IgM syndrome. AB - We report a case of an 11-year-old boy who underwent successful bone marrow transplantation for X-linked hyper-IgM syndrome (XHIM). The donor was an HLA matched brother. The patient was conditioned with busulfan, cyclophosphamide and anti-thymocyte globulin. He received 4.7 x 10(8) marrow cells per kg from the donor. Prophylaxis against graft-versus-host disease consisted of cyclosporine and short-term methotrexate. The clinical course after the bone marrow transplantation was uneventful, and 12 months after transplantation the patient was doing well with no need for therapy. We examined expression of the CD40 ligand (CD40L) on the patient's activated T lymphocytes and in vitro production of immunoglobulins by his lymphocytes. Although expression of CD40L was totally absent before the bone marrow transplant, subnormal expression appeared after the transplantation. In vitro production of IgG and IgA also was improved by the transplant. Based on our experience bone marrow transplantation appears to be a reasonable therapeutic option for patients with XHIM if HLA-matched family donors are available. PMID- 10373077 TI - Successful one antigen mismatched bone marrow transplant for chronic myeloid leukemia (CML) after two failed syngeneic transplants. AB - In May 1989, a 43-year-old woman with chronic myelocytic leukemia diagnosed in 1988 underwent a syngeneic bone marrow transplant (BMT), conditioned with cyclophosphamide-TBI while in chronic phase. Three years later, because of both cytogenetic and hematological relapse, she was treated with interferon-alpha (IFN alpha) and hydroxyurea (HU) for 3 years. In 1994 while still in chronic phase, she was conditioned with busulfan-cyclophosphamide (BU-CY) and underwent a second syngeneic BMT. In 1996, following a further cytogenetic and hematological relapse, she was again placed on IFN-alpha and HU therapy for 13 months, when she was referred to our hospital in accelerated phase. In October 1997 following thiotepa, CY and anti-thymocyte globulin conditioning, she underwent an allogeneic BMT from her 1-Ag mismatched brother. She became Ph1 negative with full chimerism and normal hematological parameters; acute graft-versus-host disease (GVHD) grade 3 of the skin and chronic GVHD of the liver occurred. At 11 months follow-up she is in good clinical condition and with a Karnofsky score of 90%. The role of a graft-versus-leukemia (GVL) effect in securing and maintaining the complete remission is discussed. PMID- 10373078 TI - Concurrent Pneumocystis carinii and cytomegalovirus pneumonia after autologous peripheral blood stem cell transplantation. AB - A 46-year-old woman developed concurrent CMV and Pneumocystis carinii pneumonia (PCP) 140 days after autologous peripheral blood stem cell transplantation (APBSCT) for AML. She was seropositive for CMV before undergoing APBSCT and had required prednisone for immune thrombocytopenia and allergic dermatitis for 9 weeks prior to the onset of pneumonia. She had also been receiving PCP prophylaxis with pentamidine aerosol every month for 3 months before developing symptoms. The pneumonia was complicated by severe hypoxia, requiring ventilator support and pneumothorax requiring chest tube thoracostomy. She recovered following treatment with trimethoprim-sulfamethoxazole (TMP-SMX), prednisone, gancyclovir and intravenous immunoglobulin. Although the overall incidence of severe CMV disease is low after APBSCT, preventive measures such as surveillance culture and secondary prophylaxis with gancyclovir may be warranted in patients whose cellular immune response is further compromised by corticosteroid use or other factors. PMID- 10373079 TI - Adoptive autoimmune hyperthyroidism following allogeneic stem cell transplantation from an HLA-identical sibling with Graves' disease. AB - Autoimmune diseases which follow allogeneic BMT from a donor who is a patient or a carrier of an autoimmune condition are considered to be a paradigm of adoptive autoimmunity. Seven cases of autoimmune thyroiditis associated with clinical hyperthyroidism have been published to date. In the case reported here a 35-year old female patient with AML of the M2 subtype received unmanipulated PBSC from her HLA-identical sister who had therapeutically controlled Graves' disease. Antithyroid antibodies, including thyrotropin receptor (TSHR) antibodies, appeared 1 year after transplant. Clinical hyperthyroidism requiring thyrostatic medication appeared after 2 years. The biological and clinical implications of adoptive, post-transplant autoimmunity are briefly discussed. PMID- 10373080 TI - Allogeneic peripheral blood stem cell transplantation for hypereosinophilic syndrome with severe cardiac dysfunction. AB - A 42-year-old male underwent an HLA-matched sibling PBSC transplant for hypereosinophilic syndrome (HES) diagnosed in August 1995. Prior to transplant he experienced progressive cardiac and pulmonary dysfunction with red cell and platelet transfusion dependence despite therapy with hydroxyurea, steroids and interferon. He received busulfan (16 mg/kg) and cyclophosphamide (120 mg/kg) as conditioning and standard GVHD prophylaxis with cyclosporin A and methotrexate. At day +336 he was transfusion independent without GVHD. Prompt reduction of the eosinophil count (<500/microl) and rapid improvement of cardiac function were documented, demonstrating the reversibility of organ dysfunction. Allogeneic PBSCT is an effective therapeutic option for patients with HES who fail conventional therapy. PMID- 10373081 TI - Therapeutic outcome assessment in permanent temporomandibular joint disc displacement. AB - In permanent temporomandibular disc displacement (TMJ-DD) outcome studies many authors claim positive effects of arthroscopic surgery, arthrocentesis and physical therapy. This literature review was undertaken to analyse whether the claimed effects are based on acceptable methodology. The recorded papers were analysed by two independent observers according to (1) method of investigation, (2) therapeutic intervention studied, (3) therapeutic outcome variables used, and (4) claimed effectiveness of the intervention. Agreement between observers was calculated. Twenty-four papers were found in which therapeutic outcome of interventions on temporomandibular disorders were studied. Six studies applied a true experimental design. Each of these six studies compared a different set of interventions. Twenty-two papers used maximal mouth opening (MMO) as an outcome variable, nine studied pain intensity on a visual analogue scale, one paper assessed the mandibular function impairment questionnaire. Kappa for overall agreement concerning the reviewing criteria was 0.82 (P < or = 0.001). No distinguishing effects on MMO, pain or function impairment were reported between arthroscopic surgery, arthrocentesis and physical therapy. Results of methodological sound outcome studies evaluating the effects of arthroscopic surgery, arthrocentesis and physical therapy are needed. PMID- 10373082 TI - Five year evaluation of class III composite resin restorations in cavities pre treated with an oxalic- or a phosphoric acid conditioner. AB - An oxalic acid solution has been proposed as a conditioning agent for resin composite restorations in two commercial adhesive systems. The durability of 163 class III restorations, including 12 class IV restorations, in cavities pre treated with an oxalic acid total etch technique or an enamel etch with phosphoric acid was studied. Each of 52 patients received at least one of each of three experimental restorations. The restorations were evaluated yearly with slightly modified United States Public Health Service (USPHS) criteria. After 5 years 95% of the restorations were evaluated as acceptable. Reasons for failure were the fracture of four fillings, including three class IV, secondary caries contiguous to two fillings and a non-acceptable colour match for one restoration. For eight class III restorations a fracture of the incisal tooth structure was registered. No differences were seen between the three experimental restorations. PMID- 10373083 TI - Wear of composite resin veneering materials in a dual-axis chewing simulator. AB - The attritional wear of human enamel and four different composite resins for the veneering of crowns was evaluated in a dual-axis chewing simulator over up to 1200000 loading cycles. Enamel showed less wear than the composite resins. However, an ultrafine compact-filled composite resin (Targis) showed a wear not statistically significantly different from that of enamel. The other composite resins showed a statistically significantly higher wear than enamel regardless whether microfine, ultrafine midway-filled or ultrafine compact-filled. PMID- 10373084 TI - In vitro cytotoxicity of dental casting alloys over 8 months. AB - Although in vitro cytotoxicity tests have been a valuable part of the biological testing of dental casting alloys, these tests are generally limited by their short-term nature ( < 168 h). The objective of the current study was to measure the in vitro cytotoxicity of representative types of dental casting alloys over a relatively long-term interval (months) and compare longer-term cytotoxicity with that seen initially. Polished casting alloy samples were exposed to cell-culture medium for 30-day intervals. During the last 3 days of each interval, the medium was changed to provide a 3-day extract which was then placed on fibroblast or macrophage cultures for 48 h. The mitochondrial activity of the cells was measured and compared to control cultures to assess the cytotoxic effect of the alloys. The cytotoxic effect was plotted versus time of medium exposure. The total time of exposure was 8 months. For most alloys, the mitochondrial response was constant over the 8 months, indicating that the cytotoxic effect of the alloys did not change significantly after extended exposure to the medium. A trend of improving biological response was suspect for a reduced-gold alloy, but this trend was not statistically significant. The mitochondrial activity of the macrophages was less sensitive to the alloy extracts than the activity of the fibroblasts, which was significantly suppressed for several alloy types. However, mitochondrial activity of the macrophages was significantly increased for several alloys in the early time intervals. The total noble metal content of the alloys was not necessarily predictive of the biological response. Thus, it appears that in this indirect test of in vitro cytotoxicity, the short-term cytotoxicity is predictive of the longer-term for many types of dental casting alloys. PMID- 10373085 TI - Fluoride release/uptake of polyacid-modified resin composites (compomers) in neutral and acidic buffer solutions. AB - The aim of the present study was to evaluate the fluoride uptake/release of polyacid-modified resin composites (compomers) in neutral and acidic buffer solutions. Two compomers (Dyract and Compoglass) were tested and the conventional glass-ionomer cement (GIC) Vivaglass Base served as a control. Forty specimens were fabricated from each of the respective materials. Twenty of these specimens were placed in artificial saliva and the other 20 specimens in a fluoridated dentifrice slurry for 5 min. Then, 10 fluoridated and 10 non-fluoridated specimens were immersed in a neutral buffer solution (pH 6.8), and the other specimens were immersed in an acidic solution (pH 4.0). After 1, 2, 3, 4, and 5 days the samples were again placed in either a fluoridated dentifrice slurry or saliva for 5 min, after which time they were transferred to fresh buffer solutions. The fluoride content of the solutions was assessed with a fluoride sensitive electrode. Fluoride release from all the materials decreased continuously during the experiment, with a significantly higher release in the acidic solution compared to the neutral buffer solution. Fluoridation did not result in an increased fluoride release for the compomers. However, the conventional GIC revealed a significantly higher fluoride release after fluoridation. It is concluded that Dyract and Compoglass can not be replenished with fluoride, irrespective of the pH value of the environment. PMID- 10373087 TI - Facial asymmetry in temporomandibular joint disorders. AB - In order to investigate skeletal deviation in patients with internal derangement of the TMJ, facial asymmetry was examined by the frontal cephalogram and compared with a control group of asymptomatic subjects. It was demonstrated that mandibular lateral displacement in the patients was significantly greater than that in the controls. The degree of displacement was significantly related to the cant of the frontal occlusal plane and the frontal mandibular plane, indicating the reduced vertical dimension of the posterior occlusal level and the ramus height on the mandibular displaced side. It is concluded that facial asymmetry due to mandibular lateral displacement is a relatively common problem in patients with internal derangement of TMJ. The cant of the frontal occlusal plane seems to be an important occlusal characteristic related to temporomandibular joint dysfunction. PMID- 10373086 TI - Shock absorbability of various restorative materials used on implants. AB - The purpose of this study was to compare, using an in vitro model, the stress absorbing ability of a microfilled composite resin and of a new low fusion ceramic (Duceram LFC) to that of gold alloy and conventional ceramic, when used as restorative materials in implant-supported prosthesis. Test crowns made of the tested materials were rigidly connected to a Branemark implant clone. The maximum amplitude of the force transmitted to the bone-implant interface, and the time to reach this amplitude were measured after applying a 100 N impact load on the occlusal surface. The gold alloy restorations transmitted the highest impact force in the shortest delay at the bone-implant interface. Microfilled composite resin Dentacolor, and low fusion ceramic Duceram LFC did not reduce the amplitude of the impact-force when compared to conventional ceramic. Nevertheless, the time to reach the maximum amplitude of this force was longer when using composite resin than when using ceramic, while Duceram LFC had no influence on this criteria. PMID- 10373088 TI - The effect of saliva on fluoride release by a glass-ionomer filling material. AB - The initial aim of this study was to investigate the effect of saliva and the formation of pellicle on the fluoride release in vitro of the glass-ionomer filling material, Chemfil Superior. For the first study glass-ionomer discs of 6 mm in diameter and 1.5 mm thick were made. Ten discs were immersed in whole stimulated saliva each day for 10 min and 10 control discs were immersed in deionized water. For the remaining 23 h and 50 min of each day, over the 20-day experimental period, both test and control discs were placed in deionized water. A considerable amount of fluoride was released on the first day (14.5 ppm F control and 13.3 ppm F test). The concentration of fluoride released on the second day fell sharply to 5.3 ppm F for controls and 4.9 ppm F for tests. This release had almost reached a plateau by day 10 and at day 20 the pellets continued to release low levels of fluoride. The concentration of fluoride released was only slightly higher for controls than for test discs when both were immersed in deionized water until day 20. However, during the 10-min period between 1.5 and 2 times as much fluoride was released into the deionized water as into saliva until day 20 when the ratio fell to 1.2:1. The second experiment assessed fluoride release when specimens were incubated for 1 h using an identical protocol. Again, less fluoride was released from the saliva-coated specimens compared with the controls (17%), which was not substantially different to the comparable 10-min samples (13%). This study indicates that saliva retards the release of fluoride from glass-ionomer and that this retarding effect is still present when discs are subsequently immersed in water compared with those that were placed in water alone. This suggests that salivary deposits have formed within minutes of immersion in saliva. This retarding effect was observed throughout the study period with the exception of the 20-day samples which had been incubated in saliva for 10 min. PMID- 10373089 TI - Influence of surface treatment on adherence energy of alloys used in bonded prosthetics. AB - Usually, shear or tensile tests are used to assess bond strengths between resin and metal. In this study, a cleavage test, the Double Cantilever Beam test, was performed to measure the adherence energy in air and in water between a 4-META resin, and five alloys (palladium, palladium-silver, gold, cobalt-chromium and nickel-chromium alloys) whose surfaces have been treated by sandblasting only or by two methods of silica coating (Silicoater MD, Rocatec) or by painting with a primer (V-Primer). Results showed that, after storage in water, it is difficult to divide the studied alloys into a dental base alloys group and a noble alloys group. Therefore, the silica coating has significantly limited the propagation of fissures in water. Higher values of adherence energy were recorded with the Rocatec system except with the palladium alloy which must be treated with the Silicoater MD system. The treatment with V-Primer was sensitive to hydrolytic attack. PMID- 10373090 TI - The stress reducing capacity of unfilled resin in a Class V cavity. AB - This study examined the stress reducing capacity of varying thicknesses of unfilled resin in a Class V cavity. A two dimensional plane strain mesh of a Class V cavity, 3 mm in diameter and 2 mm deep, was produced and the thickness of the unfilled resin layer was varied from 0 to 80 microm. A polymerization shrinkage of 1.5% was applied to the composite resin and the interfacial forces examined. The maximum shear stresses were found to occur along the pulpal floor of the restoration at the unfilled resin-dentine interface. The maximum shear stress values varied from 11.1 to 22.4 MPa and the shear stresses decreased by up to 38% as the thickness of the unfilled resin increased to 80 microm. PMID- 10373091 TI - Posture correction as part of behavioural therapy in treatment of myofascial pain with limited opening. AB - In this study, we applied cognitive behavioural intervention to subjects who had painful limited mouth opening, with or without posture correction in daily life. The efficacy of non-intervention control was then compared with it in order to study the effectiveness of posture correction as part of a biobehavioural therapy. The visual analogue scale (VAS) value of pain intensity at maximum mouth opening and disturbance in daily life sharply declined in the group which received only cognitive behavioural intervention and those who received it together with posture correction in daily life compared to the non-intervention control group although there was little difference between the intervention groups. Moreover, pain-free unassisted mouth opening was restored earlier in the group which had added posture correction. This suggests that posture correction in daily life has a positive effect in alleviating myofascial pain with limited mouth opening. PMID- 10373092 TI - The effect of bonding agents on the microleakage and bond strength of sealant in primary teeth. AB - This in vitro study investigated the effect of use of three dentine bonding agents: Scotchbond Multi-Purpose Plus (3M Dental Products, St. Paul, Minnesota, U.S.A.), Syntac (Vivadent, Schaan, Liechtenstein), Optibond Dual Cure (Kerr, Romulus, MI, U.S.A.) on microleakage and shear bond strength of a fissure sealant (Helioseal F, Vivadent, Schaan, Liechtenstein) bonded to either dry or wet (saliva contaminated) enamels of primary teeth. Newly extracted 112 non-carious primary teeth were sectioned and embedded in resin blocks. Eight groups were formed for each test. Each group consisted of 14 specimens. Group 1 and 2: fissure sealant was applied directly to etched enamel in dry and wet condition, respectively; Group 3 and 4: fissure sealant was applied onto etched and Scotch bond Multi-Purpose Plus treated enamel in dry and wet condition, respectively; Group 5 and 6: fissure sealant was applied onto etched and Syntac treated enamel in dry and wet condition, respectively; Group 7 and 8: fissure sealant was applied onto etched and Optibond Dual Cure treated enamel in dry and wet condition, respectively. The results revealed that the use of an enamel-dentine bonding agent under fissure sealant increased the bond strength and decreased the microleakage. The use of enamel-dentine bonding agents under sealant in moisture contaminated conditions gave better results than applying sealant alone onto non contaminated teeth. Finally, Scotchbond Multi-Purpose Plus yielded the best results for both tests. PMID- 10373093 TI - Influence of polymerization initiator for base monomer on microwave curing of composite resin inlays. AB - Microwave polymerization was used to make composite resin inlays and the effect examined of the concentration of polymerization initiator for the base monomer. The monomers used were 2,2-bis [4-(3-methacriloxy-2-hydroxypropoxy) phenyl] propane (Bis-GMA) and triethyleneglycol dimethacrylate (TEGDMA). Bis-GMA and TEGDMA were mixed in a ratio of 6:4 by weight and were separated into five groups. To each group was added benzoyl peroxide (BPO) in the ratios of 0.1, 0.3, 0.5, 0.7 and 0.9 wt% as the polymerization initiator. These were used as the base monomers. The results showed that the degree of conversion of the cured sample increased with increasing concentration of BPO from 0.1 to 0.5 wt%, however there was no significant difference at 0.5, 0.7 and 0.9 wt% (P> 0.01). Compression strength, diametral tensile strength and the Knoop hardness showed a similar tendency as the degree of conversion. No significant difference was recorded in the Knoop hardness between the top and the bottom surfaces (P> 0.01), which suggested a uniform polymerization in the cured sample. Thus, microwave polymerization would be an efficacious method for making resin inlays with the addition of BPO to the base monomer (Bis-GMA:TEGDMA, 6:4). The maximum conversion was found at a concentration of 0.5 wt%. PMID- 10373094 TI - Evaluation of biting performance with standardized test-foods. AB - Human masseter muscle is the main power source used for crushing or grinding the foods. Its size is regarded as an determining factor of chewing force and function. A system of measuring the masseter muscle volume from MR images was developed in this study. For a better understanding of the masseter muscle function, a standardized test-food developed in our laboratory was used. Twelve male subjects having complete dentition and healthy masticatory function were used to chew the test-foods. The EMG muscle activity of the masseter muscle during breaking and chewing the test-foods was observed. It was found that the muscle volume was positively related to the body size. The increase of the muscle activity during breaking and chewing was almost parallel to the increase of the test-food hardness, and the increase was mostly in the duration of muscle contraction. It was concluded that the measurement of masseter muscle volume is possible, and our test-foods can be used for chewing function examination with high reliability. Within a certain level of hardness, harder food requires longer muscle contraction time instead of stronger muscle force to break and chew the food. PMID- 10373095 TI - Keeping an eye on retinal clocks. AB - Circadian pacemakers that drive rhythmicity in retinal function are found in both invertebrates and vertebrates. They have been localized to photoreceptors in molluscs, amphibians, and mammals. Like other circadian pacemakers, they entrain to light, oscillate based on a negative feedback between transcription and translation of clock genes, and control a variety of physiological and behavioral rhythms that often includes rhythmic melatonin production. As a highly organized and accessible tissue, the retina is particularly well suited for the study of the input-output pathways and the mechanism for rhythm generation. Impressive advances can now be expected as researchers apply new molecular techniques toward looking into the eye's clock. PMID- 10373096 TI - Short-term rhythms of the cardiorespiratory system and their significance in neonatology. AB - Latent disturbances in the control of respiration and heart rate (HR) may be important factors in the pathogenesis of life-threatening events during infancy. A method of determining the control of the autonomic nervous system functions involves the analysis of time-dependent ultradian changes of its parameters. The breathing signal and HR variability contain rhythmic components that are generated within the cardiorespiratory network of the brain stem, through reflexes, and by feedback mechanisms. The analysis of these components may provide insights into the functioning of the cardiorespiratory control system. The prominence and precision of the rhythms are correlated with states of vigilance and underlie distinct development during the first months of life. The results of studies on infants at risk (for example, for sudden infant death), with the help of statistical and spectral analysis of time series to obtain new indices, have proved to be inconsistent in their prognostic value of thus studied parameters. Recently, the importance of qualitative and quantitative assessment of the dynamic and complex behavior of time series, based on nonlinear characteristics of the control system, has been emphasized. To what extent, however, the analysis of the dynamic behavior can be utilized for clinical purposes, such as judging the prognosis of deficiencies in control, requires further study regarding physiological baselines and the possible changes resulting from pathological states. PMID- 10373097 TI - Provisional QTL for circadian period of wheel running in laboratory mice: quantitative genetics of period in RI mice. AB - Wheel running was monitored in B x D recombinant inbred (RI) mice under dark-dark (DD) conditions, and the mean circadian period was calculated for each strain. There were significant differences for this trait among B x D recombinant inbred strains (p < .0001) and a narrow-sense heritability of 21%. Analysis of strain means and variances indicates that at least four segregating loci contribute to the genetic variance for the free-running circadian period in this population. Correlation of the strain means for the circadian period of wheel running for each RI strain against the distribution of markers at over 1500 loci along the mouse genome identified a number of provisional quantitative trait loci (QTL). There were provisional QTL for wheel running at p < .001 on chromosome 11 and at p < .01 on chromosomes 1, 6, 9, 17, and 19. Most were in agreement with a second analysis done under similar conditions. PMID- 10373098 TI - Food-entrained feeding and locomotor circadian rhythms in rats under different lighting conditions. AB - It has been suggested that two endogenous timekeeping systems, a light entrainable pacemaker (LEP) and a food-entrainable pacemaker (FEP), control circadian rhythms. To understand the function and interaction between these two mechanisms better, we studied two behavioral circadian rhythmicities, feeding and locomotor activity, in rats exposed to two conflicting zeitgebers, food restriction and light-dark cycles. For this, the food approaches and wheel running activity of rats kept under light-dark (LD) 12:12, constant darkness (DD), or constant light (LL) conditions and subjected to different scheduled feeding patterns were continuously recorded. To facilitate comparison of the results obtained under the different lighting conditions, the period of the feeding cycles was set in all three cases about 1h less than the light-entrained or free-running circadian rhythms. The results showed that, depending on the lighting conditions, some components of the feeding and wheel-running circadian rhythms could be entrained by food pulses, while others retained their free running or light-entrained state. Under LD, food pulses had little influence on the light-entrained feeding and locomotor rhythms. Under DD, relative coordination between free-running and food-associated rhythms may appear. In both cases, the feeding activity associated with the food pulses could be divided into a prominent phase-dependent peak of activity within the period of food availability and another afterward. Wheel-running activity mainly followed the food pulses. Under LL conditions, the food-entrained activity consisted mainly of feeding and wheel-running anticipatory activity. The results provide new evidence that lighting conditions influence the establishment and persistence of food entrained circadian rhythms in rats. The existence of two coupled pacemakers, LEP and FEP, or a multioscillatory LEP may both explain our experimental results. PMID- 10373099 TI - Development of inverse circadian blood pressure pattern in transgenic hypertensive TGR(mREN2)27 rats. AB - TGR(mREN2)27 (TGR) rats are transgenic animals with an additional mouse renin gene, which leads to overactivity of the renin-angiotensin system. Adult TGR rats are characterized by fulminant hypertension, hypertensive end-organ damage, and an inverse circadian blood pressure pattern. To study the ontogenetic development of cardiovascular circadian rhythms, telemetric blood pressure transmitters were implanted in male Sprague-Dawley (SPRD, n = 5) and heterozygous, transgenic TGR rats before 5 weeks of age. The TGR received either drinking water or enalapril 10 mg/L in drinking water (n = 5 per group). Drug intake was measured throughout the study by computerized monitoring of drinking volume. Circadian patterns in blood pressure and heart rate were analyzed from 5 to 11 weeks of age. In the first week after transmitter implantation, blood pressure did not differ among SPRD, untreated, and enalapril-treated TGR rats. In parallel with the rise in blood pressure of untreated TGR rats, a continuous delay of the circadian acrophase (time of fitted blood pressure maximum) was observed, leading to a complete reversal of the rhythm in blood pressure at an age of 8 weeks. Enalapril reduced blood pressure at night, but was less effective during the day, presumably due to the drinking pattern of the animals, which ingested about 90% of their daily water intake during the nocturnal activity period. After discontinuation of treatment, blood pressure returned almost immediately to values found in untreated TGR rats. In conclusion, the inverse circadian blood pressure profile in TGR rats develops in parallel with the increase in blood pressure. Direct effects of the brain renin-angiotensin system may be involved in the disturbed circadian rhythmicity in TGR(mREN2)27 rats. PMID- 10373100 TI - Urinary excretion of nitric oxide, cyclic GMP, and catecholamines during rest and activity period in transgenic hypertensive rats. AB - Dysregulation of the system of nitric oxide (NO)-cyclic 3',5'-guanosine monophosphate (cGMP) might be involved in the development of hypertension in transgenic hypertensive TGR(mREN2)27 (TGR) rats. The present study was performed to determine possible differences in the day-night pattern and the urinary excretion rates of NO and cGMP in TGR rats in comparison to normotensive Sprague Dawley (SPRD) controls. In addition, the urinary excretion of creatinine and catecholamines was measured in both rat strains. The day-night excretion patterns of NO, cGMP, catecholamines, and creatinine were preserved in TGR rats. Urinary excretion of NO was significantly decreased in TGR rats, whereas cGMP, the second messenger of NO, was elevated in the transgenic animals. Catecholamines and creatinine excretion rates did not differ between the strains. In conclusion, data suggest that a reduced NO synthesis could contribute to the increased blood pressure in the severely hypertensive rats. However, these data make it unlikely that the disturbances in the nitric oxide-cGMP system and the sympathetic nervous system are mainly responsible for the inverse circadian blood pressure rhythm in TGR rats. PMID- 10373101 TI - Timed daily administration of prolactin and corticosteroid hormone reduces murine tumor growth and enhances immune reactivity. AB - In the present study, we investigated the time-dependent interactive effects of daily injections of prolactin (PRL) and corticosterone (CORT) on the activation of lymphocyte function and inhibition of tumor growth in vivo in mice. BALB/c mice were injected subcutaneously with EMT-6 fibrosarcoma cells (a murine connective tissue tumor cell derived from mammary gland), and then different groups of animals were treated with PRL (1 microg/g body weight [BW] ip) at Oh, 4h, 8h, 12h, 16h, or 20h after CRT (1 microg/g BW ip) daily for 10 days. Different control groups were vehicle treated or treated with either hormone alone. Mice were kept in constant light 1 week before and during injections and in a 14:10 light-dark cycle thereafter. Tumor progression was monitored for up to 21 days after the cessation of treatment, and thereafter spleen lymphocytes were harvested and tested for mitogen-triggered proliferation. Prolactin administration at 8h or 16-20h after corticosteroid treatment reduced tumor volume by 77% and 49%, respectively, relative to vehicle-treated controls. Other time relations of hormone treatment were ineffectual. Further studies indicated that the immunosuppressant cyclosporin A (CSA) substantially stimulated tumor growth; this effect was completely abrogated by a simultaneous 8h related hormone treatment. How ever, the 8h hormone treatment was ineffective in inhibiting tumor growth in T-cell-deficient nude mice. Spleen lymphocytes from tumor-bearing (TB) mice showed an elevated basal proliferative capacity stimulated by concanavalin A (ConA; a stimulus for T-cell proliferation) and lipopolysaccharide (LPS; a stimulus for B-cell proliferation) compared to non-TB mice. Spleen lymphocytes from TB mice treated with CORT and PRL at 8h intervals exhibited an increased spontaneous (as well as LPS- and ConA- triggered) proliferation (by 104%, 48%, and 70%, respectively) compared with vehicle control TB mice. Fluorescence activated cell sorting (FACS) analysis of splenocytes from hormone-treated animals indicated a 34-100% increase in the CD4+ (e.g., T helper cell) population. Treatment of animals with either hormone alone did not inhibit tumor growth or stimulate immune function relative to vehicle controls. The daily rhythms of plasma PRL, CORT, and thyroxine were all substantially altered by the presence of tumor in these mice. These results indicate that appropriately timed daily treatment of PRL and CORT can attenuate tumor growth, in part, via activation of antitumor immune mechanisms. Collectively, these data suggest that circadian neuroendocrine activities must be temporally organized appropriately to inhibit tumor growth. PMID- 10373102 TI - Flow cytometric analysis of circadian changes in platelet activation using anti GMP-140 monoclonal antibody. AB - The hemostatic activity of blood shows a circadian variation with a higher frequency of acute coronary events in the morning. The thrombotic tendency of blood is influenced by many factors, including platelets. Diurnal changes of in vivo platelet activation were investigated by whole blood flow cytometry in 10 young healthy male volunteers using anti-GMP-140 (anti-alpha-granule membrane protein 140 kD) monoclonal antibody at 3h intervals from 06:00 to 24:00. We also studied circulating platelet aggregates to investigate whether there exists a similarity between the results of these methods. Results of flow cytometric analysis indicate that there is an increase in platelet activation during the period from 06:00 to 09:00. Platelet activation then decreases gradually during the period from noon to midnight. These changes are accompanied by a similar trend in circulating platelet aggregates. This suggests that GMP-140 expression on platelets is synchronized with or followed by platelet aggregate formation in vivo, and increased platelet activation may predispose individuals to thrombosis at this time. PMID- 10373103 TI - The effect of activity on the waking temperature rhythm in humans. AB - Nine healthy female subjects were studied when exposed to the natural light-dark cycle, but living for 17 "days" on a 27h day (9h sleep, 18h wake). Since the circadian endogenous oscillator cannot entrain to this imposed period, forced desynchronization between the sleep/activity cycle and the endogenous circadian temperature rhythm took place. This enabled the effects of activity on core temperature to be assessed at different endogenous circadian phases and at different stages of the sleep/activity cycle. Rectal temperature was measured at 6-minute intervals, and the activity of the nondominant wrist was summed at 1 minute intervals. Each waking span was divided into overlapping 3h sections, and each section was submitted to linear regression analysis between the rectal temperatures and the total activity in the previous 30 minutes. From this analysis were obtained the gradient (of the change in rectal temperature produced by a unit change in activity) and the intercept (the rectal temperature predicted when activity was zero). The gradients were subjected to a two-factor analysis of variance (ANOVA) (circadian phase/ time awake). There was no significant effect of time awake, but circadian phase was highly significant statistically. Post hoc tests (Newman-Keuls) indicated that gradients around the temperature peak were significantly less than those around its trough. The intercepts formed a sinusoid that, for the group, showed a mesor (+/-SE) of 36.97 (+/-0.12) and amplitude (95% confidence interval) of 0.22 degrees C (0.12 degrees C, 0.32 degrees C). We conclude that this is a further method for removing masking effects from circadian temperature rhythm data in order to assess its endogenous component, a method that can be used when subjects are able to live normally. We suggest also that the decreased effect of activity on temperature when the endogenous circadian rhythm and activity are at their peak will reduce the possibility of hyperthermia. PMID- 10373104 TI - Bright light exposure during the daytime affects circadian rhythms of urinary melatonin and salivary immunoglobulin A. AB - The effects of bright light exposure during the daytime on circadian urinary melatonin and salivary immunoglobulin A (IgA) rhythms were investigated in an environmental chamber controlled at a global temperature of 27 degrees C+/-0.2 degrees C and a relative humidity of 60%+/-5%. Seven diurnally active healthy females were studied twice, in bright and dim light conditions. Bright light of 5000 lux was provided by placing fluorescent lamps about 1 meter in front of the subjects during the daytime exposure (06:30-19:30) from 06:30 on day 1 to 10:30 on day 3. Dim light was controlled at 200 lux, and the subjects were allowed to sleep from 22:30 to 06:30 under both light exposure conditions. Urine and saliva were collected at 4h intervals for assessing melatonin and IgA. Melatonin excretion in the urine was significantly greater during the nighttime (i.e., at 06:30 on day 1 and at 02:30 on day 2) after the bright light condition than during the dim light condition. Furthermore, the concentration and the amount of salivary IgA tended to be higher in the bright light than in the dim light condition, especially during the night-time. Also, salivary IgA concentration and the total amount secreted in the saliva were significantly positively correlated with urinary melatonin. These results are consistent with the hypothesis that bright light exposure during the daytime enhances the nocturnal melatonin increase and activates the mucosal immune response. PMID- 10373105 TI - Investigation into the role of the response regulator NtrC in the metabolism and virulence of Brucella suis. AB - During infection, Brucella species have to adapt to a range of different environments. Environmental sensing in bacteria often involves the concerted action of two-component regulatory systems consisting of sensor and response regulator components. In this study, we identified, cloned and sequenced four independent response regulator gene fragments from Brucella melitensis. One amplified gene fragment showed nearly 90% identity to the response regulator subfamily of NtrC transcriptional activators, and further analysis revealed the presence of an adjacent gene encoding the sensor protein NtrB. The NtrBC two component regulatory system has been shown to play varying roles in nitrogen metabolism and potentially in virulence in other bacterial species. A B. suis ntrC isogenic mutant was constructed which showed no significant differences in growth rates compared to the wild-type strain when grown at different temperatures in vitro. However, the mutant exhibited a reduction in metabolic activity in the presence of many amino acids. The mutation did not affect survival or multiplication of B. suis in macrophages, but during the initial stages of infection in the murine brucellosis model, the ntrC mutant showed a reduced ability to multiply rapidly in splenic tissue. PMID- 10373106 TI - Mitigation emerges as major strategy for reducing losses caused by natural disasters. Board of Natural Disasters. AB - The International Decade for Natural Disaster Reduction, a United Nations program for the 1990s, focused attention on the increasing losses caused by natural hazards and promoted actions to reduce their impacts. During this period in the United States, disaster managers and other officials increased emphasis on mitigation relative to response and recovery, especially in programs of the Federal Emergency Management Agency. Many other nations and international organizations undertook similar efforts. Beyond the Decade, efforts should focus on improving risk assessments; implementing mitigation strategies; improving technologies supporting warnings and the dissemination of, and response to, warnings; improving the basis for natural disaster insurance; and assisting developing nations. PMID- 10373107 TI - Wing rotation and the aerodynamic basis of insect flight. AB - The enhanced aerodynamic performance of insects results from an interaction of three distinct yet interactive mechanisms: delayed stall, rotational circulation, and wake capture. Delayed stall functions during the translational portions of the stroke, when the wings sweep through the air with a large angle of attack. In contrast, rotational circulation and wake capture generate aerodynamic forces during stroke reversals, when the wings rapidly rotate and change direction. In addition to contributing to the lift required to keep an insect aloft, these two rotational mechanisms provide a potent means by which the animal can modulate the direction and magnitude of flight forces during steering maneuvers. A comprehensive theory incorporating both translational and rotational mechanisms may explain the diverse patterns of wing motion displayed by different species of insects. PMID- 10373108 TI - Structure of the Escherichia coli fumarate reductase respiratory complex. AB - The integral membrane protein fumarate reductase catalyzes the final step of anaerobic respiration when fumarate is the terminal electron acceptor. The homologous enzyme succinate dehydrogenase also plays a prominent role in cellular energetics as a member of the Krebs cycle and as complex II of the aerobic respiratory chain. Fumarate reductase consists of four subunits that contain a covalently linked flavin adenine dinucleotide, three different iron-sulfur clusters, and at least two quinones. The crystal structure of intact fumarate reductase has been solved at 3.3 angstrom resolution and demonstrates that the cofactors are arranged in a nearly linear manner from the membrane-bound quinone to the active site flavin. Although fumarate reductase is not associated with any proton-pumping function, the two quinones are positioned on opposite sides of the membrane in an arrangement similar to that of the Q-cycle organization observed for cytochrome bc1. PMID- 10373109 TI - Quantum computing with electrons floating on liquid helium AB - A quasi-two-dimensional set of electrons (1 < N < 10(9)) in vacuum, trapped in one-dimensional hydrogenic levels above a micrometer-thick film of liquid helium, is proposed as an easily manipulated strongly interacting set of quantum bits. Individual electrons are laterally confined by micrometer-sized metal pads below the helium. Information is stored in the lowest hydrogenic levels. With electric fields, at temperatures of 10(-2) kelvin, changes in the wave function can be made in nanoseconds. Wave function coherence times are 0.1 millisecond. The wave function is read out with an inverted dc voltage, which releases excited electrons from the surface. PMID- 10373110 TI - Spontaneous bubble domain formation in a layered ferromagnetic crystal AB - Magnetic domain structure on the surface of the layer-structured ferromagnet La1.4Sr1.6Mn2O7 was observed in the temperature range from 37 to 97 kelvin with a scanning Hall probe microscope. The sensitivity to temperature of the domain structure changes was large relative to that in conventional ferromagnets. The stable and spontaneous appearance of magnetic bubble domains without an external magnetic field was observed in the neighborhood of 70 kelvin. The phenomenon observed could provide a potential route toward magnetic bubble memory. PMID- 10373111 TI - Century-scale shifts in early holocene atmospheric CO2 concentration AB - The inverse relation between atmospheric carbon dioxide concentration and stomatal frequency in tree leaves provides an accurate method for detecting and quantifying century-scale carbon dioxide fluctuations. Stomatal frequency signatures of fossil birch leaves reflect an abrupt carbon dioxide increase at the beginning of the Holocene. A succeeding carbon dioxide decline matches the Preboreal Oscillation, a 150-year cooling pulse that occurred about 300 years after the onset of the Holocene. In contrast to conventional ice core estimates of 270 to 280 parts per million by volume (ppmv), the stomatal frequency signal suggests that early Holocene carbon dioxide concentrations were well above 300 ppmv. PMID- 10373112 TI - Contribution of disturbance to increasing seasonal amplitude of atmospheric CO2 AB - Recent increases in the seasonal amplitude of atmospheric carbon dioxide (CO2) at high latitudes suggest a widespread biospheric response to high-latitude warming. The seasonal amplitude of net ecosystem carbon exchange by northern Siberian ecosystems is shown to be greater in disturbed than undisturbed sites, due to increased summer influx and increased winter efflux. Increased disturbance could therefore contribute significantly to the amplified seasonal cycle of atmospheric carbon dioxide at high latitudes. Warm temperatures reduced summer carbon influx, suggesting that high-latitude warming, if it occurred, would be unlikely to increase seasonal amplitude of carbon exchange. PMID- 10373113 TI - Xyloglucan fucosyltransferase, an enzyme involved in plant cell wall biosynthesis. AB - Cell walls are crucial for development, signal transduction, and disease resistance in plants. Cell walls are made of cellulose, hemicelluloses, and pectins. Xyloglucan (XG), the principal load-bearing hemicellulose of dicotyledonous plants, has a terminal fucosyl residue. A 60-kilodalton fucosyltransferase (FTase) that adds this residue was purified from pea epicotyls. Peptide sequence information from the pea FTase allowed the cloning of a homologous gene, AtFT1, from Arabidopsis. Antibodies raised against recombinant AtFTase immunoprecipitate FTase enzyme activity from solubilized Arabidopsis membrane proteins, and AtFT1 expressed in mammalian COS cells results in the presence of XG FTase activity in these cells. PMID- 10373114 TI - Dissociating pain from its anticipation in the human brain. AB - The experience of pain is subjectively different from the fear and anxiety caused by threats of pain. Functional magnetic resonance imaging in healthy humans was applied to dissociate neural activation patterns associated with acute pain and its anticipation. Expectation of pain activated sites within the medial frontal lobe, insular cortex, and cerebellum distinct from, but close to, locations mediating pain experience itself. Anticipation of pain can in its own right cause mood changes and behavioral adaptations that exacerbate the suffering experienced by chronic pain patients. Selective manipulations of activity at these sites may offer therapeutic possibilities for treating chronic pain. PMID- 10373115 TI - Gating of BDNF-induced synaptic potentiation by cAMP. AB - Neurotrophins have been implicated in activity-dependent synaptic plasticity, but the underlying intracellular mechanisms remain largely unknown. Synaptic potentiation induced by brain-derived neurotrophic factor (BDNF), but not neurotrophin 3, was prevented by blockers of adenosine 3',5'-monophosphate (cAMP) signaling. Activators of cAMP signaling alone were ineffective in modifying synaptic efficacy but greatly enhanced the potentiation effect of BDNF. Blocking cAMP signaling abolished the facilitation of BDNF-induced potentiation by presynaptic activity. Thus synaptic actions of BDNF are gated by cAMP. Activity and other coincident signals that modulate cAMP concentrations may specify the action of secreted neurotrophins on developing nerve terminals. PMID- 10373116 TI - Preventing neurodegeneration in the Drosophila mutant bubblegum. AB - The Drosophila melanogaster recessive mutant bubblegum (bgm) exhibits adult neurodegeneration, with marked dilation of photoreceptor axons. The bubblegum mutant shows elevated levels of very long chain fatty acids (VLCFAs), as seen in the human disease adrenoleukodystrophy (ALD). In ALD, the excess can be lowered by dietary treatment with "Lorenzo's oil," a mixture of unsaturated fatty acids. Feeding the fly mutant one of the components, glyceryl trioleate oil, blocked the accumulation of excess VLCFAs as well as development of the pathology. Mutant flies thus provide a potential model system for studying mechanisms of neurodegenerative disease and screening drugs for treatment. PMID- 10373117 TI - Prehistoric polymers: rubber processing in ancient mesoamerica AB - Ancient Mesoamerican peoples harvested latex from Castilla elastica, processed it using liquid extracted from Ipomoea alba (a species of morning glory vine), and fashioned rubber balls, hollow rubber figurines, and other rubber artifacts from the resulting material. Chemical and mechanical analyses of the latex and of the processed rubber indicate that the enhanced elastic behavior of the rubber relative to the unprocessed latex is due to purification of the polymer component and to an increase in the strength and number of interchain interactions that are induced by organic compounds present in I. alba. These ancient peoples' control over the properties of latex and processed rubber gave rise to the Mesoamerican ball game, a central ritual element in all ancient Mesoamerican societies. PMID- 10373118 TI - Requirement for croquemort in phagocytosis of apoptotic cells in Drosophila. AB - Macrophages in the Drosophila embryo are responsible for the phagocytosis of apoptotic cells and are competent to engulf bacteria. Croquemort (CRQ) is a CD36 related receptor expressed exclusively on these macrophages. Genetic evidence showed that crq was essential for efficient phagocytosis of apoptotic corpses but was not required for the engulfment of bacteria. The expression of CRQ was regulated by the amount of apoptosis. These data define distinct pathways for the phagocytosis of corpses and bacteria in Drosophila. PMID- 10373119 TI - Vessel cooption, regression, and growth in tumors mediated by angiopoietins and VEGF. AB - In contrast with the prevailing view that most tumors and metastases begin as avascular masses, evidence is presented here that a subset of tumors instead initially grows by coopting existing host vessels. This coopted host vasculature does not immediately undergo angiogenesis to support the tumor but instead regresses, leading to a secondarily avascular tumor and massive tumor cell loss. Ultimately, however, the remaining tumor is rescued by robust angiogenesis at the tumor margin. The expression patterns of the angiogenic antagonist angiopoietin-2 and of pro-angiogenic vascular endothelial growth factor (VEGF) suggest that these proteins may be critical regulators of this balance between vascular regression and growth. PMID- 10373121 TI - Identification of both shared and distinct proteins in the major and minor spliceosomes. AB - In metazoans, two distinct spliceosomes catalyzing pre-messenger RNA splicing have been identified. Here, the human U11/U12 small nuclear ribonucleoprotein (snRNP), a subunit of the minor (U12-dependent) spliceosome, was isolated. Twenty U11/U12 proteins were identified, including subsets unique to the minor spliceosome or common to both spliceosomes. Common proteins include four U2 snRNP polypeptides that constitute the essential splicing factor SF3b. A 35-kilodalton U11-associated protein homologous to the U1 snRNP 70K protein was also identified. These data provide fundamental information about proteins of the minor spliceosome and shed light on its evolutionary relationship to the major spliceosome. PMID- 10373120 TI - Initiation of mammalian liver development from endoderm by fibroblast growth factors. AB - The signaling molecules that elicit embryonic induction of the liver from the mammalian gut endoderm or induction of other gut-derived organs are unknown. Close proximity of cardiac mesoderm, which expresses fibroblast growth factors (FGFs) 1, 2, and 8, causes the foregut endoderm to develop into the liver. Treatment of isolated foregut endoderm from mouse embryos with FGF1 or FGF2, but not FGF8, was sufficient to replace cardiac mesoderm as an inducer of the liver gene expression program, the latter being the first step of hepatogenesis. The hepatogenic response was restricted to endoderm tissue, which selectively coexpresses FGF receptors 1 and 4. Further studies with FGFs and their specific inhibitors showed that FGF8 contributes to the morphogenetic outgrowth of the hepatic endoderm. Thus, different FGF signals appear to initiate distinct phases of liver development during mammalian organogenesis. PMID- 10373122 TI - Rationing in child health services: a personal view. PMID- 10373123 TI - Pseudomonas and all that. PMID- 10373124 TI - Influence of socioeconomic conditions on growth in infancy: the 1921 Aberdeen birth cohort. AB - OBJECTIVES: To identify environmental influences on infant growth using data from a birth cohort established in 1921. DESIGN: A longitudinal cohort study. SETTING: Aberdeen 1921-22. SUBJECTS: Five hundred and sixteen individuals (263 boys and 253 girls) born in Aberdeen during 1921. Health visitor assessments ranged from two to 40 (47% received at least 10 visits). No records were available for infants who died. Individuals were grouped as those who did not breast feed, those who breast fed initially but not at 6 months, and those who were continuing to breast feed at 6 months. MAIN OUTCOME MEASURE: Rate of weight gain over the 1st year of life. A random effects model was used to identify environmental factors and conditions contributing to rate of weight gain in the 1st year of life. RESULTS: Breast feeding rates were about 80% and 50% at 10 days and 6 months, respectively. Breast fed infants were significantly heavier than bottle fed infants at 28 days but this difference disappeared by 12 months. Significant negative effects on rate of weight gain, independent of initial body weight, were found for overcrowding in family homes and maternal parity, whereas social class had no effect. CONCLUSION: Studies based on historical cohorts that have controlled socioeconomic variables only in terms of social class (derived from parental occupation) may have been subject to residual confounding. Growth in the 1st year of life is likely to reflect a number of environmental influences, some of which may continue to have effects throughout early life and beyond. PMID- 10373125 TI - Whole body bone mineral accretion in healthy children and adolescents. AB - Data on accretion in bone size and bone mineral content (BMC) are needed to evaluate bone mineralisation during childhood. Whole body bone mineral content (BMC) and bone area (BA) were determined by dual energy x ray absorptiometry (Hologic 1000/W) with a one year interval in healthy girls (n = 192) and boys (n = 140) aged 6-19 years. Annual accretion in BMC (DeltaBMC (g/year)) and BA (DeltaBA (cm2/year)) according to sex and pubertal stages were calculated. DeltaBA and DeltaBMC were highly significantly associated with pubertal stages in girls and boys. Centile curves for DeltaBA and DeltaBMC according to sex and age were constructed using the LMS method. Peak DeltaBA and DeltaBMC values were reached earlier in girls (12.3 and 12.5 years, respectively) than in boys (13.4 and 14. 2 years, respectively). The DeltaBA peak was dissociated in time from the DeltaBMC peak, indicating that increase in bone size occurs before increase in bone mineral content. Assuming that 32.2% of BMC consist of calcium, the median (90th centile) annual bone calcium accretion in pubertal stage III was 220 mg/day (302) and 317 mg/day (386) for girls and boys, respectively. To obtain an average bone calcium accretion, a high calcium absorption is needed during puberty. This may have implications for dietary calcium requirements at this time. PMID- 10373126 TI - Duodenogastric reflux: clinical and therapeutic aspects. AB - BACKGROUND: Duodenogastric reflux is believed to cause damage to gastric mucosa. Most reports on this disorder concern adult patients. PATIENTS AND METHODS: 1120 children with abdominal pain were studied; endoscopic features of duodenogastric reflux were found in 92 patients. To confirm the diagnosis of duodenogastric reflux, cholescintigraphy (Tc99-HEPIDA) was performed. Children with confirmed duodenogastric reflux by scintigraphy were given a prokinetic drug (cisapride). RESULTS: Endoscopic features of duodenogastric reflux were found in 92 children; the diagnosis was confirmed by scintigraphy in 59 patients. There was no significant difference in the severity of inflammation in gastric mucosa compared with the control group, whereas significantly fewer of these patients were infected with Helicobacter pylori. There was no correlation between regions of isotope accumulation and inflammatory lesions in the stomach. The prokinetic drug (cisapride) helped eliminate or greatly reduce duodenogastric reflux in children. CONCLUSIONS: When endoscopic features of duodenogastric reflux are found the final diagnosis should be based on an examination that does not itself influence the motility of the gastrointestinal tract: cholescintigraphy seems to be a useful method. However, because the use of milk as a test meal affects the scintigraphic image, there was no correlation between the area of isotope accumulation and the localisation of inflammatory lesions in the stomach. Duodenogastric reflux seems to be less important as a cause of inflammatory lesions than other factors (such as genetic predisposition, stress, etc). Prokinetic drugs have a beneficial influence on treatment results in children with inflammatory lesions of gastric mucosa with duodenogastric reflux. PMID- 10373128 TI - FETAL AND NEONATAL EDITION july issue PMID- 10373129 TI - Feeding difficulties and foregut dysmotility in Noonan's syndrome. AB - PURPOSE: Noonan's syndrome is a common dysmorphic syndrome in which failure to thrive and gastrointestinal symptoms are frequent but poorly understood. DESIGN: Twenty five children with Noonan's syndrome were investigated by contrast radiology, pH monitoring, surface electrogastrography (EGG), and antroduodenal manometry (ADM). RESULTS: Sixteen had poor feeding and symptoms of gastrointestinal dysfunction. All 16 required tube feeding. Seven of 25 had symptoms of foregut dysmotility and gastro-oesophageal reflux. In the most symptomatic children (four of seven) EGG showed fasting frequency gradient loss along the stomach fundus and pylorus with antral postprandial frequency loss. ADM showed shortened fasting cycle length, with abnormal phase III and shortened postprandial activity containing phasic contractions. IMPLICATIONS: Gastroduodenal motor activity was reminiscent of 32-35 week preterm patterns. The feeding difficulties appear to resolve as gut motility matures. In Noonan's syndrome, feeding problems appear to be the result of delayed gastrointestinal motor development. PMID- 10373127 TI - Chronic intestinal pseudo-obstruction: treatment and long term follow up of 44 patients. AB - AIMS: To document the long term course of chronic idiopathic intestinal pseudo obstruction syndrome (CIIPS) in children with defined enteric neuromuscular disease, and the place and type of surgery used in their management; in addition, to identify prognostic factors. METHODS: Children with CIIPS were investigated and treated prospectively. RESULTS: Twenty four children presented congenitally, eight during the 1st year of life, and 10 later. Twenty two had myopathy and 16 neuropathy (11 familial). Malrotation was present in 16 patients, 10 had short small intestine, six had non-hypertrophic pyloric stenosis, and 16 had urinary tract involvement. Thirty two patients needed long term parenteral nutrition (TPN): for less than six months in 19 and for more than six months in 13, 10 of whom are TPN dependent; 14 are now enteral feeding. Prokinetic treatment improved six of 22. Intestinal decompression stomas were used in 36, colostomy relieved symptoms in five of 11, and ileostomy in 16 of 31. A poor outcome (death (14) or TPN dependence (10)) was seen with malrotation (13 of 16), short small bowel (eight of nine), urinary tract involvement (12 of 16), and myopathic histology (15 of 22). CONCLUSIONS: In CIIPS drugs are not helpful but decompression stomas are. Outcome was poor in 24 of 44 children (15 muscle disorder, 10 nerve disease). PMID- 10373131 TI - Inflicted head injury-"could do better" PMID- 10373130 TI - Improved outcome of acute myeloid leukaemia in Down's syndrome. AB - OBJECTIVE: To review the clinical features, treatment, and outcome of children in the UK with Down's syndrome and acute myeloid leukaemia (AML). DESIGN: A retrospective study of 59 children with Down's syndrome and AML presenting between 1987 and 1995. Data were obtained from hospital case notes, trial records, and by questionnaire. RESULTS: The patients were unusually young (median age, 23 months) with a predominance of megakaryoblastic AML. Two of the seven infants who presented with abnormal myelopoesis aged 2 months or younger achieved complete spontaneous remission. Most of the older children with AML (32 of 52) were treated on recognised intensive protocols but 13 received individualised treatment and seven symptomatic treatment alone. Only four received a bone marrow transplant (BMT) in first remission. For the 45 older children who received chemotherapy the overall survival was 55% (median follow up 4.5 years). Patients on individualised protocols had a similar overall survival and toxic death rate but marginally higher relapse rate than those on standard (intensive) protocols. Children with Down's syndrome treated on the national AML 10 trial had a similar overall survival (70% v 59%) at five years to children of comparable age without Down's syndrome: their improved relapse risk (12% v 38%) offset the slight increase in deaths as a result of treatment toxicity (19% v 11%). CONCLUSION: Neonates with Down's syndrome and abnormal myelopoesis may achieve spontaneous remission, and older children with Down's syndrome and AML can be treated successfully with intensive chemotherapy, without BMT. PMID- 10373133 TI - Inclusion benefit PMID- 10373132 TI - Persistent nocturnal cough: randomised controlled trial of high dose inhaled corticosteroid. AB - OBJECTIVE: To investigate the effect of a short course of inhaled corticosteroid in the treatment of isolated and persistent nocturnal cough in children. DESIGN: Randomised double blind placebo controlled study. SETTING: Subjects' homes in east London, England. SUBJECTS: Consecutively referred children, 1-10 years old, with persistent nocturnal cough. INTERVENTIONS: Placebo or fluticasone propionate 1 mg twice daily for three nights and 500 microg twice daily for 11 nights. Videotaping of children at night: two nights' baseline, nights 3 and 4 after three days of inhaled corticosteroid, and nights 15 and 16. MAIN OUTCOME MEASURE: A fall in 75% of coughs from baseline. RESULTS: 50 subjects were recruited. The median number of coughs in the baseline period for the inhaled corticosteroid group and placebo group were 92 and 71, respectively (p = 0.43) and, on nights 15 and 16, 8 and 36, respectively (p < 0. 01). Compared to baseline, both groups of subjects improved significantly by nights 15 and 16 (p < 0.01; p < 0.01). Comparing the inhaled corticosteroid and placebo groups, coughs fell to a median of 22% and 57% of baseline totals on nights 3 and 4, respectively (p = 0.38), and 8% and 35% on nights 15 and 16, respectively (p = 0.02). 17 of 24 subjects on inhaled corticosteroid who completed the study and 8 of 23 on placebo improved by 75% after two weeks (p = 0.03). CONCLUSIONS: Children with persistent nocturnal cough improve in two weeks after referral on placebo. There is a modest benefit from a two week course of high dose inhaled corticosteroid. PMID- 10373134 TI - Effect of formoterol on clinical parameters and lung functions in patients with bronchial asthma: a randomised controlled trial. AB - AIMS: To determine the role of formoterol in the treatment of children with bronchial asthma who are symptomatic despite regular use of inhaled corticosteroids. METHODS: A randomised, double blind, parallel group, placebo controlled study to investigate the effects of inhaled formoterol (12 microg twice a day) in 32 children with moderate to severe bronchial asthma. The study consisted of two week run in periods and six week treatment periods, during both of which the patients continued their regular anti-inflammatory drugs. The efficacy parameters were symptom scores, bronchodilator use, daily peak expiratory flow rates (PEFR), methacholine hyper-reactivity, forced expiratory volume in one second (FEV1), lung volumes, and airway conductance. RESULTS: Formoterol treatment for six weeks decreased symptom scores, PEFR variability, and the number of rescue salbutamol doses, and increased morning and evening PEFR significantly. No adverse reactions were seen. CONCLUSION: These findings suggest that inhaled formoterol is effective in controlling chronic asthma symptoms in children who are symptomatic despite regular use of inhaled corticosteroids. PMID- 10373135 TI - Malnutrition and growth failure in cyanotic and acyanotic congenital heart disease with and without pulmonary hypertension. AB - AIM: To investigate the effect of several types of congenital heart disease (CHD) on nutrition and growth. PATIENTS AND METHODS: The prevalence of malnutrition and growth failure was investigated in 89 patients with CHD aged 1-45 months. They were grouped according to cardiac diagnosis: group aP (n = 26), acyanotic patients with pulmonary hypertension; group ap (n = 5), acyanotic patients without pulmonary hypertension; group cp (n = 42), cyanotic patients without pulmonary hypertension; and group cP (n = 16), cyanotic patients with pulmonary hypertension. Information on socioeconomic level, parental education status, birth weight and nutrition history, number of siblings, and the timing, quality, and quantity of nutrients ingested during weaning period and at the time of the examination were obtained through interviews with parents. RESULTS: There was no significant difference between groups in terms of parental education status, socioeconomic level, duration of breast feeding, and number of siblings (p > 0.05). Group cP patients ingested fewer nutrients for their age compared to other groups. 37 of the 89 patients were below the 5th centile for both weight and length, and 58 of 89 patients were below the 5th centile for weight. Mild or borderline malnutrition was more common in group aP patients. Most group cp patients were in normal nutritional state, and stunting was more common than wasting. Both moderate to severe malnutrition and failure to thrive were more common in group cP patients. CONCLUSION: Patients with CHD are prone to malnutrition and growth failure. Pulmonary hypertension appears to be the most important factor, and cyanotic patients with pulmonary hypertension are the ones most severely affected. This study shows the additive effects of hypoxia and pulmonary hypertension on nutrition and growth of children with CHD. PMID- 10373136 TI - Follow up of precocious pseudopuberty associated with isolated ovarian follicular cysts. AB - The clinical outcomes of seven girls presenting with pseudosexual precocity caused by isolated autonomous ovarian follicular cysts are presented. Six of the seven girls, aged 11 months to 6.9 years, had a unilateral ovarian cyst detected by ultrasound at the first acute episode. Plasma oestradiol was raised in only five of the cases, but all had a low response to luteinising hormone releasing hormone stimulation. Follow up lasted for up to eight years with recurrent episodes of variable frequency and severity in all seven patients. Evidence of McCune-Albright syndrome appeared later in only three patients. It could not be predicted from the initial symptoms or the clinical course. Mutations of the G(s)alpha protein leading to activation were investigated in the lymphocytes and ovarian and bone tissues of four patients. Only one patient showed a mutation in bone tissue. Close follow up with repeated searches for skeletal lesions remains necessary since the distribution of somatic mutations cannot be assessed by molecular studies. Most patients with recurrent ovarian cysts require a conservative approach. PMID- 10373137 TI - Effect of DDAVP on nocturnal enuresis in a patient with nephrogenic diabetes insipidus. AB - The case of an 8 year old boy with both nocturnal enuresis and nephrogenic diabetes insipidus is presented. Diagnosis of nephrogenic diabetes insipidus was based on a typical medical history, the characteristic result of a fluid restriction test, the lack of an effect of 1-desamino-8-D-arginine (DDAVP) on both urine osmolality and plasma coagulation factors and, finally, the detection of a hemizygous missense mutation within the arginine vasopressin (AVP) receptor gene. Hydrochlorothiazide treatment and dietary measures reduced the patient's urine volume to one third of its original volume. However, this had no effect on enuresis. The daily intranasal application of DDAVP did not further reduce urine output but dramatically decreased the frequency of bed wetting. This observation contradicts the common notion that the therapeutic effect of DDAVP in nocturnal enuresis is the result of compensation for a nocturnal AVP deficit. Rather, it points to a different mode of action of DDAVP in patients with enuresis. It is hypothesised that central AVP receptors are a target of DDAVP and that they might play an important role in the pathogenesis of nocturnal enuresis. PMID- 10373138 TI - Outbreak of Escherichia coli O157 in a nursery: lessons for prevention. AB - OBJECTIVES: To identify risk factors for transmission of verocytotoxin producing Escherichia coli O157 (VTECO157) and means of prevention. STUDY DESIGN: Outbreak investigation: retrospective cohort study. SETTING: A nursery (child care centre) in North Wales. SUBJECTS: Children attending (n = 104). METHODS: Faeces were examined using sorbitol MacConkey agar (SMAC), with cefixime, tellurite, and rhamnose; enrichment in modified tryptone soya broth; and immunomagnetic separation. Symptoms and exposure data were obtained from questionnaires to parents/guardians and children's toiletting and feeding records kept at the nursery. MAIN OUTCOME MEASURE: A "case" was defined as a child with verocytotoxin producing E coli O157 isolated from faeces, or a history of haemolytic uraemic syndrome (HUS) and antibodies to E coli O157 lipopolysaccharide, during the period 10 August to 30 September 1995. RESULTS: The attack rate was 31 in 104. Two children developed HUS. There were higher attack rates among girls and friends who played together. Cases were more likely to attend the nursery more frequently. The mean number of recorded bowel motions/child/half day was 0.51 in cases and 0.21 in well children. Child to staff ratios were high preceding and during the outbreak. CONCLUSIONS: A sick child is the most plausible source of infection with subsequent person to person transmission. The record of children's toiletting discriminated between cases and well children and might have allowed earlier detection of the outbreak. This simple record could be considered by other child care facilities as a means of giving early warning of problems with infectious intestinal diseases. PMID- 10373139 TI - Vulvovaginitis: clinical features, aetiology, and microbiology of the genital tract. AB - AIM: To clarify the contribution of clinical and environmental factors and infection to the aetiology of vulvovaginitis in premenarchal girls, and to determine clinical indicators of an infectious cause. DESIGN: It was necessary first to define normal vaginal flora. Cases were 50 premenarchal girls > 2 years old with symptoms of vulvovaginitis; 50 controls were recruited from girls in the same age group undergoing minor or elective surgery. RESULTS: Interview questionnaire showed no difference between cases and controls in regards to hygiene practices, exposure to specific irritants, or history of possible sexual abuse. Normal vaginal flora was similar to that described in previous studies, with the exception of organisms likely to be associated with sexual activity. 80% of cases had no evidence of an infectious cause. In the 10 cases in whom an infectious cause was found, there was significantly more visible discharge and distinct redness of the genital area on examination compared with other cases. CONCLUSIONS: The findings suggest that vulvovaginitis in this age group is not usually infectious or necessarily related to poor hygiene, specific irritants or sexual abuse, although any of these can present with genital irritation. The possibility of sexual abuse should always be considered when a child presents with genital symptoms, but our data indicate it is not a common contributing factor. Infection is generally associated with vaginal discharge and moderate or severe inflammation. PMID- 10373141 TI - Plastic migration from implanted central venous access devices. AB - BACKGROUND: This is the first reported study of histologically confirmed migration from intravenous access devices in children. METHODS: The capsules from around intravenous access devices were examined by light microscopy to determine the extent of the foreign body response; energy dispersive x ray analysis was performed to document the elemental content of the foreign material. RESULTS: A fibroconnective tissue capsule was found around all the samples. Elemental silicon was found in six of 13 tissue samples, and a foreign body giant cell reaction was seen in three of these. CONCLUSIONS: The pseudocapsule that surrounds an implanted vascular access device often has residual foreign material, including silicone. PMID- 10373140 TI - Dual versus triple therapy of Helicobacter pylori infection: results of a multicentre trial. AB - OBJECTIVE: To compare dual therapy (omeprazole and amoxicillin) with triple therapy (omeprazole, amoxicillin, and clarithromycin) in the treatment of Helicobacter pylori infection. The efficacy of 1 mg/kg/day omeprazole was randomly compared with 2 mg/kg/day. STUDY DESIGN: 252 patients (median age, 11.0 years; range, 3-18) presenting with chronic abdominal pain underwent endoscopy and a 13C-urea breath test. Gastric biopsy specimens were taken for histological examination and for the rapid urease test. Patients were treated for two weeks: group A (n = 63) received amoxicillin (50 mg/kg; maximum, 2 g/day), group B (n = 73) received amoxicillin and clarithromycin (20 mg/kg; maximum, 1 g/day). Both groups were randomly treated with either 1 or 2 mg/kg omeprazole (maximum, 80 mg/day). Diagnostic procedures were repeated four weeks after the end of treatment. RESULTS: 11 patients were excluded; 136 patients were H pylori positive (56%), 105 of whom were re-examined after treatment. Helicobacter pylori was eradicated in 52% of group A and 83% of group B. The dose of omeprazole had no influence on the eradication rate. Specificity and sensitivity of the rapid urease test were 94% and 93%, respectively. Specificity and sensitivity of the 13C-urea breath test were 93% and 95%, respectively. CONCLUSIONS: Dual therapy can no longer be recommended. Triple therapy is more effective than dual therapy in the eradication of H pylori infection. The lower dose of 1 mg/kg omeprazole was as effective as 2 mg/kg. PMID- 10373142 TI - Role of the oxygen uptake efficiency slope in evaluating exercise tolerance. AB - OBJECTIVE: To investigate the interprotocol agreement of oxygen uptake efficiency slope (OUES). METHODS: 16 Japanese children and adolescents (10 boys and six girls) underwent two sessions of maximal exercise testing according to the following two treadmill protocols: the standard Bruce protocol and the rapidly increasing staged (RIS) protocol. Maximal oxygen uptake (VO2max), the ventilatory anaerobic threshold (VAT), and the OUES were obtained from the gas analysis data. Agreement between the protocols was tested by means of the Bland-Altman method. RESULTS: Interprotocol agreement was excellent for the OUES (limit of agreement, 18% to 17% of the mean value), slightly less good for VO2max (limit of agreement, -20% to 24% of the mean value), and poor for the VAT (limit of agreement, -31% to 31% of the mean value). CONCLUSION: These results confirm the clinical usefulness of the OUES as a measure of evaluating exercise tolerance in the paediatric population. PMID- 10373143 TI - Teenagers with epilepsy. PMID- 10373144 TI - Dietary products used in infants for treatment and prevention of food allergy. Joint Statement of the European Society for Paediatric Allergology and Clinical Immunology (ESPACI) Committee on Hypoallergenic Formulas and the European Society for Paediatric Gastroenterology, Hepatology and Nutrition (ESPGHAN) Committee on Nutrition. PMID- 10373145 TI - Stroke in childhood. PMID- 10373146 TI - Pharmacological advance in the treatment of acute brain injury. PMID- 10373147 TI - Primary health care guide to common UK parasitic diseases PMID- 10373148 TI - Allergy and allergic diseases-The new mechanisms and therapeutics PMID- 10373149 TI - Environment and health. PMID- 10373150 TI - Research priorities in environmental health. PMID- 10373151 TI - Moving beyond journals: the future arrives with a crash. PMID- 10373152 TI - Junior doctors: waving or drowning? PMID- 10373153 TI - New antidepressants for old people? PMID- 10373154 TI - Parkinson's disease genetics comes of age. PMID- 10373155 TI - Doctors advised to take special care with human albumin. PMID- 10373156 TI - New research demolishes link between MMR vaccine and autism. PMID- 10373157 TI - Dutch regulate sponorship of hospitals. PMID- 10373158 TI - Clinton proposes coverage of prescription drugs by Medicare. PMID- 10373159 TI - In brief PMID- 10373160 TI - AMA's scheme to protect JAMA's independence has drawbacks. PMID- 10373161 TI - "Wrong" women taking HRT. PMID- 10373162 TI - HRT raises risk of rare, treatable breast cancer PMID- 10373163 TI - England launches campaign on teenage pregnancies. PMID- 10373164 TI - Restoring medical services in Kosovo will be a massive task. PMID- 10373165 TI - The carnage wrought by major economic change: ecological study of traffic related mortality and the reunification of Germany. AB - OBJECTIVE: To document the effects of sudden economic change on death rates for occupants of cars in the former German Democratic Republic (East Germany). DESIGN: Ecological time series study of East Germany in comparison with the former Federal Republic of Germany (West Germany) before and after reunification in 1990. SETTING: East and West Germany from 1985 to 1996. SUBJECTS: Populations of East and West Germany between 1985 and 1996. MAIN OUTCOME MEASURES: Death rates for occupants of cars. RESULTS: After the reunification of Germany, East Germany experienced a sudden, temporary affluence and a concomitant fourfold increase in death rates for car occupants between 1989 and 1991. Although death rates increased in all age groups, young adults (aged 18-24) were most affected. The death rate per 100 000 population for those aged 18-20 years increased 11 fold between 1989 and 1991; for those aged 21-24 years the increase was eightfold. CONCLUSION: A tragic consequence of the reunification of Germany was a dramatic increase in the death rate for car occupants. Sudden economic change and availability of cars resulted in both a rise in vehicle ownership and an increase in the number of inexperienced drivers on roads that were ill prepared for the increased traffic. The lesson learnt from Germany is that during times of economic change and modernisation, measures to prevent the predictable injury deaths that will result need to be considered. PMID- 10373166 TI - Prevention of vertical transmission of HIV: analysis of cost effectiveness of options available in South Africa. AB - OBJECTIVE: To assess the cost effectiveness of vertical transmission prevention strategies by using a mathematical simulation model. DESIGN: A Markov chain model was used to simulate the cost effectiveness of four formula feeding strategies, three antiretroviral interventions, and combined formula feeding and antiretroviral interventions on a cohort of 20 000 pregnancies. All children born to HIV positive mothers were followed up until age of likely death given current life expectancy and a cost per life year gained calculated for each strategy. SETTING: Model of working class, urban South African population. RESULTS: Low cost antiretroviral regimens were almost as effective as high cost ones and more cost effective when formula feeding interventions were added. With or without formula feeding, low cost antiretroviral interventions were likely to save lives and money. Interventions that allowed breast feeding early on, to be replaced by formula feeding at 4 or 7 months, seemed likely to save fewer lives and offered poorer value for money. CONCLUSIONS: Antiretroviral interventions are probably cost effective across a wide range of settings, with or without formula feeding interventions. The appropriateness of formula feeding was highly cost effective only in settings with high seroprevalence and reasonable levels of child survival and dangerous where infant mortality was high or the protective effect of breast feeding substantial. Pilot projects are now needed to ensure the feasibility of implementation. PMID- 10373167 TI - Universal HIV screening of pregnant women in England: cost effectiveness analysis. AB - OBJECTIVE: To estimate the cost effectiveness of universal, voluntary HIV screening of pregnant women in England. DESIGN: Cost effectiveness analysis. Cost estimates of caring for HIV positive children were based on the stage of HIV infection and calculated using data obtained from a London hospital between 1986 and 1996. These were combined with estimates of the health benefits and costs of antenatal screening so that the cost effectiveness of universal, voluntary antenatal screening for HIV infection in England could be estimated. MAIN OUTCOME MEASURES: Lifetime, direct costs of medical care of childhood HIV infection; life years gained as a result of the screening programme; net cost per life year gained for different pretest counselling costs; and different prevalence rates of pregnant women who were unaware that they were HIV positive. RESULTS: Estimated direct lifetime medical and social care costs of childhood HIV infection were pound178 300 using a 5% discount rate for time preference (1995-6 prices). In high prevalence areas screening pregnant women for HIV is estimated to be a cost effective intervention with a net cost of less than pound4000 for each life year gained. For areas with comparatively low prevalence rates, cost effectiveness could be less than pound20 000 per life year gained, depending on the number of pregnant women who are unaware that they are infected and local screening costs. CONCLUSIONS: Our results confirm recent recommendations that universal, voluntary antenatal HIV screening should be implemented in the London area. Serious consideration of the policy should be given for other areas in England depending on local prevalence and screening costs. PMID- 10373169 TI - Unimpressed PMID- 10373168 TI - Antenatal HIV testing: assessment of a routine voluntary approach. PMID- 10373170 TI - Predicting who develops chronic low back pain in primary care: a prospective study. AB - OBJECTIVES: To quantify the relative contribution of premorbid and episode specific factors in determining the long term persistence of disabling symptoms of low back pain. DESIGN: Prospective cohort study. SETTING: Two general practices in the south Manchester area. PARTICIPANTS: 180 patients, who previously participated in a cross sectional population survey, who consulted because of low back pain during the study period. They were followed at 1 week and 3 and 12 months after consultation. MAIN OUTCOME MEASURE: Persistent disabling low back pain in the 12 months after the consultation. RESULTS: Disabling low back pain persisted in one third of participants after consultation and was more common with increasing age, among those with a history of low back pain, and in women. Persistence of symptoms was associated with "premorbid" factors (high levels of psychological distress (odds ratio 3.3; 95% confidence interval 1.5 to 7.2), poor self rated health (3.6; 1.9 to 6.8), low levels of physical activity (2.8; 1.4 to 5.6), smoking (2. 1; 1.0 to 4.3), dissatisfaction with employment (2.4; 1.3 to 4.5)) and factors related to the episode of low back pain (duration of symptoms, pain radiating to the leg (2.6; 1.3 to 5.1), widespread pain (6.4; 2.7 to 15), and restriction in spinal mobility). A multivariate model based on six factors identified groups whose likelihood of persistent symptoms ranged from 6% to 70%. CONCLUSIONS: The presence of persistent low back pain is determined not only by clinical factors associated with pain but also by the premorbid state. PMID- 10373171 TI - Statistics notes: variables and parameters. PMID- 10373173 TI - Lesson of the week: importance of distinguishing between cellulitis and varicose eczema of the leg. PMID- 10373172 TI - Recent advances: paediatric surgery. PMID- 10373174 TI - ABC of intensive care: respiratory support. PMID- 10373175 TI - Water and health in Europe: an overview. PMID- 10373177 TI - Pride before a fall PMID- 10373176 TI - Climate change and human health in Europe. PMID- 10373178 TI - A different route to health: implications of transport policies. PMID- 10373180 TI - The virus and the hookworm PMID- 10373179 TI - Food production and food safety. PMID- 10373181 TI - World bank must do more to develop safe and sustainable transportation systems. PMID- 10373182 TI - The paradoxes of genetically modified foods. Protection of the public health should underpin all decisions. PMID- 10373184 TI - Current census categories are not a good match for identity. PMID- 10373183 TI - Preventing osteoporosis, falls, and fractures among elderly people. Few exercise programmes studied have prevented falls. PMID- 10373185 TI - Importance of health economics must be recognised when trials are designed. PMID- 10373186 TI - Audit of use of ACE inhibitors and monitoring in general practice. Guidelines on monitoring, on their own, are not sufficient. PMID- 10373187 TI - Advice given to patients with fractures. Drug treatments that reduce fracture rate are underused after vertebral fractures. PMID- 10373188 TI - Strategy to reformulate waiting lists. New Zealand has some suggestions for NHS priority system for elective surgery. PMID- 10373189 TI - Real-time priority scoring system must be used for prioritisation on waiting lists. PMID- 10373190 TI - Eligibility criteria improve children's access to long term ventilation at home. PMID- 10373191 TI - Maybe the time has come for the primacy of the patient in the NHS. PMID- 10373192 TI - Edinburgh college's consensus statements are not purely for UK. PMID- 10373193 TI - South Africa must admit that HIV/AIDS is its greatest enemy. PMID- 10373195 TI - Public health conference PMID- 10373194 TI - Arthur levin PMID- 10373196 TI - The white death: A history of tuberculosis PMID- 10373197 TI - Essential subtleties on the silver Sea PMID- 10373198 TI - Evidence based practice in primary care PMID- 10373199 TI - Bookcase PMID- 10373200 TI - These people are like you and me PMID- 10373202 TI - The environment: everybody's business PMID- 10373201 TI - Postcard from sussex PMID- 10373203 TI - Death rates for car occupants in east germany increased greatly after reunification PMID- 10373204 TI - Model shows that incidence of HIV infection in children can be reduced PMID- 10373205 TI - Universal antenatal HIV screening is cost effective in areas of high prevalence PMID- 10373206 TI - Routine HIV tests in pregnancy are acceptable to women PMID- 10373207 TI - Persistent back pain can be predicted PMID- 10373208 TI - Europe must manage the health effects of inevitable climate change PMID- 10373209 TI - Council honorees and the nobel prize : our continued anniversary celebration PMID- 10373210 TI - Seven lessons from two candidate genes in human essential hypertension: angiotensinogen and epithelial sodium channel. AB - The candidate gene approach to understanding the genetics of human essential hypertension is discussed by analyzing the contribution of 2 genes, angiotensinogen (AGT) and epithelial amiloride-sensitive sodium channel (ENaC). From a large series of studies conducted in humans and animals, it appears that the AGT gene plays a significant but modest role in human blood pressure variance. Mutations of the beta- and gamma-ENaC subunits are responsible for Liddle's syndrome, but the implication of the 3 ENaC subunits in essential hypertension is still questionable. Several lessons can be learned from these studies and applied to other candidate genes in essential hypertension: (1) Many linkage or association studies have a limited statistical power; (2) The genetic findings may vary greatly according to the populations studied; (3) There is a need for better phenotyping of the hypertensive population; (4) The causal relationship between molecular variants and hypertension is and will be difficult to establish firmly; (5) The contribution of genetic studied in rodents to the molecular genetics of human hypertension must be re-examined; (6) Most molecular variants lead to a low attributable risk in the population or a low individual effect at the individual level; and (7) It is too early to propose dietary recommendations and specific drug treatment according to patients' genotypes. PMID- 10373211 TI - Linkage analysis of candidate genes and gene-gene interactions in chinese hypertensive sib pairs. AB - Previous studies of hypertension in humans and experimental animal models have identified a number of candidate genes that have since been implicated as possibly contributing to essential hypertension. Among them are the genes encoding angiotensinogen, renin, the beta- and gamma-subunits of the epithelial sodium channel (beta/gamma-ENaC), alpha-adducin, and kallikrein (KLK). To examine the role of possible contribution of these genes in ethnic Chinese, as well as the epistatic interaction among them, we studied a large cohort of hypertensive sib pairs from China. DNA samples from 310 concordant affected sibling pairs with hypertension were tested for linkage with the use of excess allele-sharing algorithms based on genotyping with highly informative GT-repeat microsatellite markers localized in the immediate vicinity of the genes encoding angiotensinogen, renin, beta- and gamma-ENaC, alpha-adducin, and KLK. Affected sib pair analysis conducted according to 3 different methods (Statistical Analysis for Genetic Epidemiology [S.A.G.E. ]/SIBPAL, MAPMAKER/SIBS, and affected pedigree member [APM] methods) revealed no evidence for linkage of any of these genes to primary hypertension in the population studied. Moreover, 2-locus sib pair linkage analyses to test for gene-gene interactions among each possible pair of candidate genes failed to yield any statistically significant results. Our findings provide no support for a significant contribution of the angiotensinogen, renin, beta/gamma-ENaC, alpha-adducin, or KLK genes, alone or in concert, to the pathogenesis of essential hypertension among Chinese. Our results emphasize the possible role of ethnic differences for complex disease genetics, as well as the need for large, well-characterized investigations. PMID- 10373212 TI - Factor V Leiden and thermolabile methylenetetrahydrofolate reductase gene variants in an East Anglian preeclampsia cohort. AB - Preeclampsia is a heritable condition that develops as a result of widespread vascular endothelial dysfunction. The thrombotic tendency in this condition has suggested a number of candidate genes, and there have been recent reports of positive association with the Leiden variant of factor V and the thermolabile variant of methylenetetrahydrofolate reductase. We attempted to reproduce these results in a large cohort of well-characterized women with preeclampsia, recruited prospectively within the East Anglian region of the United Kingdom. Women in the preeclampsia cohort (n=283) were genotyped for both the Leiden variant (G1691A) of factor V and the thermolabile variant (C677T) of methylenetetrahydrofolate reductase. Genotype and allele frequencies were compared with those of 2 control groups, one consisting of women recruited prospectively (n=100) from the same maternity hospital as the subjects and another consisting of normotensive women (n=100) from East Anglia. No significant differences were detected. Specifically, the carrier rate for the Leiden variant was 5.3% in the preeclampsia group and 5. 5% in the combined control group. T677 homozygotes for methylenetetrahydrofolate reductase were 11% and 11.5% in the 2 groups, respectively. We conclude that there is no evidence of association of preeclampsia with either of these 2 polymorphisms in our study population. PMID- 10373213 TI - Variable renal atrial natriuretic factor gene expression in hypertension. AB - We have previously established the existence of atrial natriuretic factor (ANF) gene expression within the renal parenchyma. Neither the role nor the regulation of this extracardiac source of ANF is clearly defined. To determine whether renal ANF gene expression, similar to cardiac expression, is linked to the activity of the renin-angiotensin system (RAS), we compared renal ANF gene expression in rats after suprarenal aortic banding, a hypertension model associated with activation of RAS, and in the deoxycorticosterone acetate (DOCA)-salt model, which is characterized by depression of RAS. Renal ANF mRNA was measured with a quantitative competitive reverse transcription polymerase chain reaction method. DOCA-salt hypertension significantly reduced the expression of renal ANF. In contrast, aortic banding significantly increased renal ANF expression. In both cases, ANF gene expression in the heart increased. Ramipril treatment at 10 micrograms/kg of aortic-banded rats, a treatment that specifically affects local RAS but maintains hypertension, normalized renal ANF mRNA levels. Altogether, these results suggest that renal ANF gene expression is modulated by local RAS and is independent of circulating RAS and hypertension per se. The marked decrease of renal ANF mRNA in DOCA-salt hypertension suggests a pathogenic role for renal ANF gene downregulation by decreasing the sodium excretory mechanism mediated by the local expression of ANF acting on receptors found in the inner medullary collecting ducts. In aortic banding, renal ANF gene expression upregulation suggests a local compensatory function consistent with the consensus role of natriuretic peptides in the modulation of RAS, thus ameliorating the sodium-retaining effects of renal underperfusion. PMID- 10373215 TI - Superoxide anion production is increased in a model of genetic hypertension: role of the endothelium. AB - The hypothesis that the decreased nitric oxide (NO) availability observed in spontaneously hypertensive stroke-prone rats (SHRSP) is due to excess superoxide (O2-) was examined. O2- generation, measured by lucigenin chemiluminescence, was studied in 12- to 16-week male and female Wistar-Kyoto rats (WKY) and SHRSP. In addition, expression of the gene encoding endothelial NO synthase, the enzyme involved in NO generation, was investigated. O2- generation was increased in male and female SHRSP (4.11+/-0.24 and 3. 84+/-0.28 nmol O2-. min-1. mg-1 respectively) compared with their WKY counterparts and was significantly higher in male than female WKY (1.22+/-0.08 in males and 0.8+/-0.08 nmol O2-. min-1. mg 1 respectively) (SHRSP versus WKY P<0.0001, 95% CI -3.39, -2.51; male versus female WKY P=0.0029, 95% CI -0.67, -0.17). Removal of the endothelium by rubbing or addition of NO synthase inhibitors attenuated O2- generation in SHRSP but not WKY. In males, removal of the endothelium reduced O2- generation from 3.86+/-0.12 to 1.35+/-0. 08 nmol. min-1. mg-1 (P<0.0001, 95% CI 2.29, 2.81), whereas addition of L-NAME caused a reduction from 4.13+/-0.17 to 1.32+/-0.16 nmol. min-1. mg-1 (P<0.0001, 95% CI 2.36, 2.83). Similar reductions were observed in females. L arginine had no significant effect, but tetrahydrobiopterin significantly decreased O2- generation in SHRSP from 4.04+/-0.11 to 2.36+/-0.40 nmol. min-1. mg 1 (P=0.0026, 95% CI 0.89, 2.44). Endothelial NO synthase mRNA expression was significantly greater in SHRSP than in WKY and in WKY males than in WKY females. These results show that O2- generation is increased in SHRSP and that the tissue and enzymatic sources of this excess O2- appear to be the endothelium and eNOS, respectively. The increase in O2- generation could explain the decreased availability of basal NO observed in this model of genetic hypertension. PMID- 10373214 TI - p53-mediated upregulation of BAX gene transcription is not involved in Bax-alpha protein overexpression in the left ventricle of spontaneously hypertensive rats. AB - An association of increased apoptosis with overexpression of the proapoptotic protein Bax-alpha has been reported in the left ventricle of adult spontaneously hypertensive rats (SHR). Both alterations were corrected in SHR that received long-term treatment with the AT1 antagonist losartan. To gain insight into the regulation of cardiac Bax-alpha protein in genetic hypertension, we investigated the expression of the protein p53 (a BAX gene transcription factor) and BAX mRNA in the left ventricle of 30-week-old Wistar-Kyoto rats (WKY), SHR, and SHR treated with losartan (20 mg. kg-1. d-1) during 14 weeks before death. The expression of p53 and Bax proteins was assessed by Western blot analysis. The expression of BAX mRNA was assessed by Northern blot analysis. The density of apoptotic cells was assessed by direct immunoperoxidase detection of biotin labeled deoxyuridine nucleotides. Compared with WKY, untreated SHR exhibited increased apoptosis (P<0.05), increased Bax-alpha protein (P<0.05), and similar levels of p53 protein and BAX mRNA. Losartan given long term was associated with the normalization of apoptosis and Bax-alpha protein expression. The expression of BAX mRNA was decreased (P<0. 05) in treated SHR compared with untreated SHR. No changes in the expression of p53 protein were observed in losartan-treated SHR. These results suggest that overexpression of the Bax-alpha protein seen in the left ventricle of adult SHR with increased apoptosis is not related to a p53 mediated upregulation of BAX gene transcription. Our data also suggest that normalization of Bax-alpha protein observed in SHR after long-term blockade of angiotensin II type 1 receptors may be due to the inhibition of BAX gene transcription. PMID- 10373216 TI - Enhanced blood pressure variability in eNOS knockout mice. AB - It has been shown previously that endogenous nitric oxide can buffer arterial blood pressure variability in dogs and rats. In these former studies, all isoforms of the nitric oxide synthase were blocked pharmacologically and an increased blood pressure variability was observed. Thus the question as to which isoform of the nitric oxide synthase is responsible for the blood pressure buffering effect of endogenous nitric oxide remains unraveled. In the present study, we therefore compared blood pressure variability in knockout mice that lack specifically the gene for endothelial nitric oxide synthase with their respective wild-type controls. One day after carotid artery cannulation, blood pressure was recorded in these conscious mice. During resting conditions, blood pressure variability was markedly enhanced in knockout mice compared with wild type mice (10.5+/-1.5 mm Hg2 vs 6.0+/-0.8 mm Hg2, P<0.05). Power spectral analysis revealed that this increase in blood pressure variability is manifested at low frequencies that range from 0.05 to 0.40 s-1 (Hz) (5.1+/-1.0 mm Hg2 vs 2.5+/-0.5 mm Hg2, P<0.05). On the basis of these results, we conclude that the blood pressure buffering effect of endogenous nitric oxide is mediated by the endothelial isoform of the nitric oxide synthase. In addition, endothelial nitric oxide is most effective in buffering blood pressure oscillations at frequencies that range from 0.05 to 0.40 s-1 (Hz) in conscious mice. PMID- 10373217 TI - Cortisol effects on body mass, blood pressure, and cholesterol in the general population. AB - The effects of excess cortisol secretion on blood pressure and fat deposition are well documented, but the importance of this glucocorticoid in controlling these processes in normal individuals is less clear. We studied the relationship between cortisol excretion rate (tetrahydrocortisol [THF]+allo THF+tetrahydrocortisone [THE]) and a range of important cardiovascular risk factors in 439 normal subjects (238 male) sampled from the North of Glasgow (Scotland) population. There were marked gender differences: female subjects were lighter and had lower blood pressures and cortisol levels, whereas HDL cholesterol was higher. The pattern of cortisol metabolism was also different; the index of 11beta-hydroxysteroid dehydrogenase activity (THF+allo-THF/THE) was lower and that of 5alpha-reductase (allo-THF/THF) was higher. There was a strong correlation of blood pressure (positive), cholesterol (positive), and HDL cholesterol (negative in women, positive in men) with age. Cortisol excretion rate did not correlate with blood pressure but correlated strongly with parameters of body habitus (body mass index and waist and hip measurements [positive]) and HDL cholesterol (negative). With multiple regression analysis, there remained a significant association of cortisol excretion rate with HDL cholesterol in men and women and with body mass index in men. These results suggest that glucocorticoids regulate key components of cardiovascular risk. PMID- 10373218 TI - Maternal hypertension and progeny blood pressure: role of aldosterone and 11beta HSD. AB - Epidemiological and experimental evidence suggests that gestational events modulate the level of blood pressure that will be "normal" for the individual as an adult. Glucocorticoid excess during gestation is associated with low birth weight, a large placenta, and adult hypertension in humans and animals. It has been proposed that the deficiency in placental 11beta-hydroxysteroid dehydrogenase activity in humans produces a gestational hormonal milieu, notwithstanding normal circulating levels of glucocorticoids, that predisposes the adult progeny to hypertension. Animal studies indicate that maternal hypertension, excess glucocorticoids, and hydroxysteroid dehydrogenase inhibition program adult blood pressure. Blood pressures of Sprague-Dawley rat dams were manipulated during gestation with continuous intracerebroventricular infusions of vehicle, aldosterone, 11alpha-hydroxyprogesterone, or carbenoxolone at doses known to produce hypertension with no renal effects or with subcutaneous infusions of larger, equally hypertensinogenic doses that produce systemic effects. Blood pressures of all treated dams were significantly greater (P<0.01) during gestation than those of the vehicle ICV control rats but not significantly different from each other. The blood pressures of both male and female progeny (n>/=6 per group, comprising representatives from at least 4 litters) were measured after 6 weeks of age. No significant difference was found in the blood pressure of the pups regardless of the maternal gestational blood pressure or treatment with an enzyme inhibitor, even after high-salt diet challenge. PMID- 10373219 TI - Angiotensin-converting enzyme inhibition restores hepatocyte growth factor production in patients with congestive heart failure. AB - Endothelium-dependent vasodilation is impaired in patients with congestive heart failure. For vascular endothelium, hepatocyte growth factor (HGF) is one of the most potent and specific growth factors, which acts protectively against endothelial dysfunction. HGF production is downregulated by angiotensin II (Ang II) in vitro. We hypothesized that HGF production is impaired as the result of increased Ang II in patients with congestive heart failure, and that if so, the impaired production should be restored with angiotensin-converting enzyme inhibitors (ACE-I). We studied 16 patients with congestive heart failure caused by previous anterior myocardial infarction in whom left ventricular ejection fraction was 35+/-8% (mean+/-SD). Before and approximately 4 weeks after the treatment with ACE-I, blood samples were collected to measure the levels of HGF, Ang II, and brain natriuretic peptide as a biochemical marker for severity of heart failure. We also studied 5 control subjects, in whom heparin increased HGF production to 48+/-5-fold. However, in patients with heart failure, HGF response to heparin was significantly attenuated (24+/-5-fold, P<0.05 vs control). Therapy with ACE-I decreased the levels of Ang II and brain natriuretic peptide and restored HGF production in response to heparin by 43+/-7-fold, comparable to the control response. In conclusion, impaired HGF production was restored after the treatment with ACE-I probably by the mechanism of Ang II suppression. This novel effect of ACE-I may contribute to the clinical improvement in patients with heart failure and thereby may have an important therapeutic implication. PMID- 10373220 TI - Therapeutic angiogenesis induced by human recombinant hepatocyte growth factor in rabbit hind limb ischemia model as cytokine supplement therapy. AB - Hepatocyte growth factor (HGF) exclusively stimulates the growth of endothelial cells without replication of vascular smooth muscle cells and acts as a survival factor against endothelial cell death. Therefore we hypothesized that a decrease in local vascular HGF might be related to the pathogenesis of peripheral arterial disease. We initially evaluated vascular HGF concentration in the vessels of patients with arteriosclerosis obliterans. Consistent with in vitro findings that hypoxia downregulated vascular HGF production, vascular HGF concentration in the diseased segments of vessels from patients with arteriosclerosis obliterans was significantly decreased as compared with disease-free segments from the same patients (P<0.05), accompanied by a marked reduction in HGF mRNA. On the other hand, a novel therapeutic strategy for ischemic diseases that uses angiogenic growth factors to expedite and/or augment collateral artery development has recently been proposed. Thus in view of the decreased endogenous vascular HGF, rhHGF (500 micrograms/animal) was intra-arterially administered through the internal iliac artery of rabbits in which the femoral artery was excised to induce unilateral hind limb ischemia, to evaluate the angiogenic activity of HGF, which could potentially have a beneficial effect in hypoxia. Administration of rhHGF twice on days 10 and 12 after surgery produced significant augmentation of collateral vessel development on day 30 in the ischemic model as assessed by angiography (P<0.01). Serial angiograms revealed progressive linear extension of collateral arteries from the origin stem artery to the distal point of the reconstituted parent vessel in HGF-treated animals. In addition, we examined the feasibility of intravenous administration of rhHGF in a moderate ischemia model. Importantly, intravenous administration of rhHGF also resulted in a significant increase in angiographic score as compared with vehicle (P<0.01). Overall, a decrease in vascular HGF might be related to the pathogenesis of peripheral arterial disease. In the presence of decreased endogenous HGF, administration of rhHGF induced therapeutic angiogenesis in the rabbit ischemic hind limb model, as potential cytokine supplement therapy for peripheral arterial disease. PMID- 10373222 TI - Age-related abnormalities in arterial compliance identified by pressure pulse contour analysis: aging and arterial compliance. AB - The objective of this study was to evaluate age-related changes in pulsatile arterial function. Aging alters arterial pulsatile function and produces consistent changes in the pressure pulse contour. A reduced systemic arterial compliance that can be derived from analysis of the pulse contour is regarded as the best clinical index of impaired pulsatile arterial function and may mark the presence of early vascular damage. We analyzed intra-arterial brachial artery waveforms in 115 healthy normotensive volunteers (83 men, 32 women) and radial artery waveforms obtained with the use of a calibrated tonometer device in 212 healthy volunteers (147 women, 65 men). A computer-based assessment of the diastolic pressure decay and a modified Windkessel model of the circulation were used to quantify changes in arterial waveform morphology in terms of large artery or capacitive compliance, oscillatory or reflective compliance in the small arteries, inertance, and systemic vascular resistance. Large artery compliance and oscillatory compliance correlated negatively with age for both invasive and noninvasive groups (r=-0.50 and r=-0.55; r=-0.37 and r=-0.66; P<0.001 for all). The slopes of the regression lines for the decline in oscillatory compliance with age were significantly steeper than those recorded for large artery compliance estimates. The change in blood pressure with age independently contributed to the decrease in large artery compliance but not oscillatory compliance in both groups. Consistent age-related changes were found in the pressure pulse contour by analysis of waveforms obtained invasively or noninvasively from the upper limb. The change in the oscillatory or reflective compliance estimate was independent of blood pressure change and may represent a better marker than large artery or capacitive compliance of the degenerative aging process in altering pulsatile arterial function. PMID- 10373221 TI - Muscular strength training is associated with low arterial compliance and high pulse pressure. AB - Aerobic exercise training increases arterial compliance and reduces systolic blood pressure, but the effects of muscular strength training on arterial mechanical properties are unknown. We compared blood pressure, whole body arterial compliance, aortic impedance, aortic stiffness (measured by beta-index and carotid pulse pressure divided by normalized systolic expansion [Ep]), pulse wave velocity, and left ventricular parameters in 19 muscular strength-trained athletes (mean+/-SD age, 26+/-4 years) and 19 sedentary controls (26+/-5 years). Subjects were healthy, non-steroid-using, nonsmoking males, and athletes had been engaged in a strength-training program with no aerobic component for a minimum of 12 months. There was no difference in maximum oxygen consumption between groups, but handgrip strength (mean+/-SEM, 44+/-2 versus 56+/-2 kg; P<0.01) and left ventricular mass (168+/-8 versus 190+/-8 g; P<0.05) were greater in athletes. Arterial stiffness was higher in athletes, as evidenced by lower whole body arterial compliance (0.40+/-0.04 versus 0.54+/-0.04 arbitrary compliance units; P=0.01), higher aortic characteristic impedance (1.55+/-0.13 versus 1.18+/-0.08 mm Hg. s. cm-1; P<0.05), beta-index (4.6+/-0.2 versus 3.8+/-0.4; P<0. 05), and ln Ep (10.86+/-0.06 versus 10.60+/-0.08; P<0.01). Femoral-dorsalis pedis pulse wave velocity was also higher in the athletes, but carotid-femoral pulse wave velocity was not different. Furthermore, both carotid (56+/-3 versus 44+/-2 mm Hg; P<0.001) and brachial (60+/-3 versus 50+/-2 mm Hg; P<0.01) pulse pressures were higher in the athletes, but mean arterial pressure and resting heart rate did not differ between groups. These data indicate that both the proximal aorta and the leg arteries are stiffer in strength-trained individuals and contribute to a higher cardiac afterload. PMID- 10373223 TI - Increased chymase-dependent angiotensin II formation in human atherosclerotic aorta. AB - Locally formed angiotensin II (Ang II) and mast cells may participate in the development of atherosclerosis. Chymase, which originates from mast cells, is the major Ang II-forming enzyme in the human heart and aorta in vitro. The aim of the present study was to investigate aortic Ang II-forming activity (AIIFA) and the histochemical localization of each Ang II-forming enzyme in the atheromatous human aorta. Specimens of normal (n=9), atherosclerotic (n=8), and aneurysmal (n=6) human aortas were obtained at autopsy or cardiovascular surgery from 23 subjects (16 men, 7 women). The total, angiotensin-converting enzyme (ACE) dependent, and chymase-dependent AIIFAs in aortic specimens were determined. The histologic and cellular localization of chymase and ACE were determined by immunocytochemistry. Total AIIFA was significantly higher in atherosclerotic and aneurysmal lesions than in normal aortas. Most of AIIFA in the human aorta in vitro was chymase-dependent in both normal (82%) and atherosclerotic aortas (90%). Immunocytochemical staining of the corresponding aortic sections with antichymase, antitryptase or anti-ACE antibodies showed that chymase-positive mast cells were located in the tunica adventitia of normal and atheromatous aortas, whereas ACE-positive cells were localized in endothelial cells of normal aorta and in macrophages of atheromatous neointima. The density of chymase- and tryptase-positive mast cells in the atherosclerotic lesions was slightly but not significantly higher than that in the normal aortas, and the number of activated mast cells in the aneurysmal lesions (18%) was significantly higher than in atherosclerotic (5%) and normal (1%) aortas. Our results suggest that local Ang II formation is increased in atherosclerotic lesions and that chymase is primarily responsible for this increase. The histologic localization and potential roles of chymase in the development of atherosclerotic lesions appear to be different from those of ACE. PMID- 10373224 TI - Mechanistic differences of various AT1-receptor blockers in isolated vessels of different origin. AB - The functional inhibitory characteristics of the angiotensin II type 1 receptor blockers (ARB) candesartan; irbesartan; and losartan and its active metabolite EXP 3174 (EXP) were studied in rabbit aortic strips and rat portal vein preparations in vitro. Moreover, plasma-protein binding was determined, and the binding was high (>98. 5%) for all ARBs. These values were needed to relate the concentrations of the ARBs used in vitro to the nonprotein bound concentrations in clinical use. In both vascular preparations, candesartan caused a marked decrease in the maximal contractile response of the angiotensin II (Ang II) concentration-response curve. Losartan, EXP, and irbesartan caused a rightward parallel shift without any major effects on the maximal response to Ang II. The inhibitory effect of candesartan developed slowly (maximal effect after >30 minutes) and lasted >2 hours despite repeated washing of the vessels. The effect of losartan, irbesartan, and EXP had a faster onset, and most of the inhibitory effect disappeared after washing. The duration of the inhibitory effects of the ARBs were not related to lipophilicity of the compounds. Cooling of the rat portal vein preparations to 4 degrees C before administration of candesartan prevented the persistent inhibition of Ang II response seen at 37 degrees C. For the other ARBs studied, the magnitude of inhibition and the speed of recovery of the Ang II response were independent of the incubation temperature before washing. In addition, when candesartan was given to conscious rats, the inhibitory effect on Ang II-induced blood pressure responses persisted during the 24-hour period despite nondetectable plasma concentrations of candesartan at 24 hours. It is concluded that functional inhibitory characteristics of candesartan differ from those of the other ARBs tested. At clinically relevant concentrations, candesartan is an insurmountable and long-lasting antagonist of the vascular contractile responses to Ang II. PMID- 10373225 TI - AT2 receptor and vascular smooth muscle cell differentiation in vascular development. AB - The angiotensin II type 2 (AT2) receptor is transiently expressed at late gestation in the fetal vasculature, but its expression rapidly declines after birth. We have previously demonstrated that the expression of this receptor mediates decline in vascular DNA synthesis that occurs at this stage of vascular development. To examine further the role of the AT2 receptor in vasculogenesis, we have focused on the effect of the AT2 receptor on vascular smooth muscle cell (VSMC) differentiation. In this study, we examined the time-dependent expression of differentiation markers for VSMCs in the aorta of wild-type and AT2 receptor null mice. alpha-Smooth muscle actin was expressed at the early stage of differentiation and exhibited unchanged expression before and after the peak of AT2 receptor expression, which was observed at embryonic day 20, neonatal day 1, and thereafter. No difference in alpha-smooth muscle actin expression was observed between the wild-type and AT2 receptor-null mice. In contrast, the mRNA levels for calponin, expressed in the late stage of VSMC differentiation, were significantly higher in the wild-type mouse aorta as compared with the AT2 receptor-null mice, which correlates with expression of the AT2 receptor. Moreover, the protein levels of calponin and high-molecular-weight caldesmon (h caldesmon) showed lower expression in the aorta of AT2 receptor knockout mice at 2 and 4 weeks after birth. Taken together, our results suggest that the AT2 receptor promotes vascular differentiation and contributes to vasculogenesis. PMID- 10373226 TI - Stimulation of the renin-angiotensin system by endothelin subtype A receptor blockade in conscious dogs. AB - Previous studies in dogs have shown additive or even synergistic effects of combined angiotensin-converting enzyme inhibition and either nonselective endothelin subtype A/B (ETA/B) or selective endothelin subtype A (ETA) receptor blockade on renal vascular resistance and mean arterial blood pressure. A possible mechanism underlying this interaction may be a stimulation of the renin angiotensin system during endothelin (ET) receptor blockade. We therefore investigated whether plasma renin activity and renin release are regulated by the ETA receptor. Experiments were made in conscious, chronically instrumented dogs receiving either saline or the selective ETA receptor antagonist LU 135252 (10 mg/kg IV). Eighty to 100 minutes after the administration of LU 135252 (n=5), heart rate (99+/-7 versus 81+/-6 bpm; P<0.05) and renal blood flow (327+/-40 versus 278+/-36 mL/min; P<0.05) were increased significantly, whereas mean arterial blood pressure tended to be lower (93+/-5 versus 105+/-7 mm Hg). These changes were associated with a 2-fold increase in plasma renin activity (0.74+/ 0.12 versus 0.37+/-0.10 ng angiotensin I per milliliter per hour; P<0.05). Measurements of renin release at various renal perfusion pressures (n=5) with the use of a vascular occluder implanted around the left renal artery revealed that ETA receptor blockade did not alter renin release at resting renal perfusion pressure (78+/-25 versus 71+/-39 U/min) but strongly enhanced the sensitivity of pressure-dependent renin release <80 mm Hg approximately 2.2-fold. In conclusion, selective ETA receptor blockade is associated with a stimulation of the circulating renin-angiotensin system, which results from both a sensitization of pressure-dependent renin release and a larger proportion of blood pressure values falling into the low pressure range, where renin release is stimulated. These find-ings strengthen the view that ET and the renin-angiotensin system closely interact to regulate vascular resistance and provide a physiological basis for synergistic hypotensive effects of a combined blockade of both pressor systems. PMID- 10373227 TI - beta-2 Adrenergic receptor variants affect resting blood pressure and agonist induced vasodilation in young adult Caucasians. AB - Recent evidence suggests that the prodownregulatory Gly16 allele of the beta-2 adrenergic receptor (beta-2 AR) is associated with essential hypertension in African Caribbeans. To further investigate the effect of the glycine (Gly)16 and arginine (Arg)16 beta-2 AR variants on hemodynamics, we investigated the agonist mediated in vivo vasodilation in normotensive Austrian Caucasians and analyzed the results with respect to the Gly16/Arg16 polymorphism. Fifty-seven normotensive men, 20 to 32 years of age with body mass index of 18.7 to 29.9 kg/m2, were genotyped for the Arg16/Gly16 beta-2 AR alleles. All 15 Gly16/Gly16 subjects, all 12 Arg16/Arg/16 subjects, and 27 of 30 heterozygous subjects underwent hemodynamic measurements while supine after an overnight fast. The observers were unaware of the subjects' genotypes. The subjects received a graded infusion of the selective beta-2 AR agonist salbutamol (0.07, 0.14, and 0.21 microgram/kg per minute, respectively), each dose over 8 minutes. Stroke volume and blood pressure were determined continuously by means of impedance cardiography and oscillometry, respectively. The last 4 minutes of each infusion were evaluated statistically. Basal mean blood pressure was higher in the Gly16/Gly16 subjects compared with Arg16/Arg16 subjects (mean+/-SD: 81.6+/-6.14 versus 75.2+/-4.93 mm Hg, P<0.01). Homozygous Gly16 subjects showed a significantly decreased vasodilation during the first dose of salbutamol infusion compared with Arg16/Arg16 subjects (Deltatotal peripheral resistance index 17.9+/-14.4 versus -30. 6+/-8.3%, P<0.01) despite increased sympathetic counterregulation in the Arg16/Arg16 group (Deltaheart rate +16.9+/-7.0% versus +8.6+/-7. 0%, P<0.01; Deltacardiac index +39.5+/-18.5% versus 21.4+/-18.8%, P<0.05). Our results provide additional evidence that the Gly16/Arg16 alleles of the beta-2 AR are intimately related to blood pressure regulation and deserve further studies in the pathogenesis of essential hypertension. PMID- 10373228 TI - Bradykinin stimulates tissue plasminogen activator release in human vasculature. AB - Bradykinin stimulates tissue plasminogen activator (tPA) release in isolated perfused animal tissues. The present study tests the hypothesis that bradykinin increases tPA release in humans through local effects on the vasculature. Graded doses of sodium nitroprusside (0.8 to 3.2 micrograms/min), acetylcholine (ACh) (7.5 to 60 micrograms/min), and bradykinin (100 to 400 ng/min) were administered intra-arterially in random order in 10 salt-depleted (10 mmol/d of Na) normotensive volunteers. None of the drugs altered mean arterial pressure or heart rate. Forearm blood flow (FBF) was measured by strain-gauge plethysmography. All 3 drugs caused a dose-dependent increase in FBF, although ACh was less potent than either nitroprusside or bradykinin (maximum FBF 7.5+/ 2.4 versus 10.0+/-1.5 and 11.9+/-2.1 mL. 100 mL-1. min-1, respectively). Bradykinin caused a significant, dose-dependent increase in venous (effect of dose F=9. 9, P=0.028 by ANOVA), but not arterial (F=0.154, P=0.92) tPA antigen in the infused arm. Thus, net tPA release increased significantly in response to bradykinin (50.6+/-13.3 at the highest dose versus 0. 9+/-0.4 ng. 100 mL-1. min 1 at baseline, P=0.014). In contrast, bradykinin did not affect plasminogen activator inhibitor antigen. Neither nitroprusside nor ACh altered plasma levels of tPA or plasminogen activator inhibitor antigen. Bradykinin increased tPA release across the forearm in the absence of systemic effects. This effect could not be attributed to changes in blood flow because doses of equivalent potency of the vasodilator nitroprusside did not increase tPA. These data demonstrate that bradykinin stimulates tPA release in the human vasculature. PMID- 10373229 TI - An enhanced effect of arginine vasopressin in bradykinin B2 receptor null mutant mice. AB - Under water restriction, arginine vasopressin (AVP) is released and promotes water reabsorption in the distal nephron, mainly through AVP V2-receptors. It has been proposed that renal kinins counteract the hydro-osmotic effect of AVP. We hypothesized that kinins acting through B2 receptors antagonize the urinary concentrating effect of AVP. To test this, bradykinin B2 receptor knockout mice (B2-KO) and 129/SvEv mice (controls) were placed in metabolic cages and urine collected for 24 hours (water ad libitum). After that, urine was again collected from the same mice during 24 hours of water restriction. Urinary volume (UV), urinary osmolarity (UOsm), and urinary Na+ (UNaV) and K+ (UKV) excretion were determined. On water restriction, UV in controls decreased by approximately 25%, whereas in B2-KO mice there was almost a 60% drop in urinary output (P=0.001 versus controls). In the controls, water restriction increased UOsm by 347 mOsm/kg H2O, approximately 14% above baseline (NS), whereas in knockout mice the increase was 3 times that seen in the controls: >1000 mOsm/kg H2O (P=0.001 versus controls). Compared with normohydration, UNaV and UKV in the water-restricted state increased more in controls than in B2-KO mice. This difference in electrolyte excretion could be explained by greater dehydration in the controls (dehydration natriuresis). In a second protocol, we tried to mimic the effect of endogenous AVP by exogenous administration of an AVP V2-receptor agonist, desmopressin (DDAVP). To suppress endogenous AVP levels before DDAVP administration, mice were volume-overloaded with dextrose and alcohol. UOsm was 685+/-125 and 561+/-58 mOsm/kg H2O in water-loaded controls and B2-KO mice, respectively. After DDAVP was injected subcutaneously at a dose of 1 microgram/kg, UOsm increased to 1175+/-86 mOsm/kg H2O (Delta+490 mOsm) in the controls and 2347+/-518 mOsm/kg H2O (Delta+1786 mOsm) in B2-KO mice (P<0.05 versus controls). We concluded that water restriction or exogenous administration of an AVP V2-receptor agonist has a greater urinary concentrating effect in B2-KO mice than in controls, suggesting that endogenous kinins acting through B2 receptors oppose the antidiuretic effect of AVP in vivo. PMID- 10373230 TI - Effect of ceramide on KCa channel activity and vascular tone in coronary arteries. AB - A sphingomyelin metabolite, ceramide, serves as a second messenger in a variety of mammalian cells. Little is known regarding the production and actions of this novel intracellular signaling lipid molecule in the vasculature. The present study was designed to test the hypothesis that a ceramide-mediated signaling pathway is present in coronary arterial smooth muscle and that ceramide serves as an inhibitor of the large-conductance Ca2+-activated potassium (KCa) channels and mediates vasoconstriction in coronary circulation. We found that C2-ceramide produced a concentration-dependent decrease in KCa channel activity in vascular smooth muscle cells from small bovine coronary arteries. The average channel activity of the KCa channels in cell-attached patches decreased from 0.046+/-0.01 to 0. 008+/-0.001 at a C2-ceramide concentration of 10 micromol/L. In inside-out patches, C2-ceramide (1 micromol/L) reduced the average channel activity of the KCa channels from 0.06+/-0.007 to 0.016+/-0. 004. Dithiothreitol, an inhibitor of acidic sphingomyelinase (1 mmol/L), increased the average channel activity of the KCa channels in cell-attached patches from 0.05+/-0.02 of control to 0.26+/-0.04, a 5-fold increase that was reversed by addition of 1 micromol/L ceramide. Glutathione, an inhibitor of neutral sphingomyelinase, was without effect. C2 ceramide significantly reduced the diameter of isolated perfused small coronary arteries in a concentration-dependent manner. Addition of 1 micromol/L C2 ceramide decreased average arterial diameter by 28%. When 14C-sphingomyelin was incubated with coronary arterial homogenates at pH 7.4 and pH 5. 0, 14C-choline phosphate and ceramide were produced. The conversion rates of 14C-sphingomyelin into 14C-choline phosphate and ceramide were 65.1+/-1.0 fmol/min per milligram protein at pH 7.4 and 114. 6+/-8.3 fmol/min per milligram protein at pH 5.0. We conclude that both acidic and neutral sphingomyelinases are present in the bovine coronary arteries and that ceramide inactivates the KCa channel in arterial smooth muscle cells and hence exerts a tonic vasoconstrictor action in coronary microcirculation. PMID- 10373231 TI - Effects of amlodipine and cilnidipine on cardiac sympathetic nervous system and neurohormonal status in essential hypertension. AB - N-Type calcium channel antagonists may suppress sympathetic activity. The purpose of this study was to assess the effects of amlodipine and cilnidipine on the cardiac sympathetic nervous system and the neurohormonal status of essential hypertension. 123I-metaiodobenzylguanidine (MIBG) cardiac imaging was performed and blood samples were taken to determine plasma renin activity and plasma norepinephrine concentration before and 3 months after drug administration in 47 patients with mild essential hypertension. Twenty-four of the patients were treated with 5 to 10 mg/d of amlodipine; the other 23 were treated with 10 to 20 mg/d of cilnidipine. For comparison, 12 normotensive subjects were also studied. No significant differences were found in the basal characteristics between the 2 hypertensive groups. In both hypertensive groups, both the systolic and diastolic blood pressures were significantly reduced to similar levels 3 months after drug treatment. Before the drug treatment, the 2 hypertensive groups had a significantly higher washout rate and lower heart-to-mediastinum (H/M) ratio compared with the normotensive subjects. The H/M ratio significantly increased (P<0.05) in combination with a decreased washout rate (P<0.02) after drug treatment in the cilnidipine group. In the amlodipine group, a significant decrease in washout rate (P<0. 04) was noted, without an increase in the H/M ratio. However, no significant changes were found in plasma renin activity and plasma norepinephrine concentration in either group. Thus, in patients with essential hypertension, cilnidipine suppressed cardiac sympathetic overactivity and amlodipine had a little suppressive effect. Cilnidipine may provide a new strategy for treatment of cardiovascular diseases with sympathetic overactivity. PMID- 10373233 TI - High stress responsivity predicts later blood pressure only in combination with positive family history and high life stress. AB - High cardiovascular responsivity to stressors has not consistently improved prediction of later blood pressure increases beyond the predictive effects of baseline pressure. Animal models suggest that genetic susceptibility to hypertension and frequent stress exposure are important modulating factors in stress-related hypertension. Thus in 103 men originally tested at age 18 to 22 years and reassessed 10 years later, interactive effects of genetic susceptibility (defined as 1 or more hypertensive parents) with high stress responsivity (defined as top 25% on the basis of blood pressure and cardiac responses during both reaction time and cold pressor tasks) were examined in relation to follow-up systolic and diastolic levels and to change in blood pressure status from normal (diastolic<80 mm Hg) to marginally elevated (diastolic 85 to 95 mm Hg). Men with the combination of high stress response and hypertensive parents demonstrated higher systolic (P<0.05) and diastolic levels (P<0.05) at follow-up, and they showed a 7-fold increase (7.5, 95% confidence intervals 2.3, 24.3; P<0.001) in relative risk of change in blood pressure status versus men with no family history and a 3-fold increase (3.8, confidence intervals 1.5, 9.6; P<0.004) versus less stress-responsive men who also had hypertensive parents. In 65 men who also provided ratings of daily stress, family historyxstress responsivityxdaily stress interactions were significant in predicting follow-up systolic and diastolic levels (P<0.006 and 0.03, respectively), with highest pressure levels seen when high life stress was reported by high stress responders and/or men with hypertensive parents. In conclusion, results suggest that stress responsivity as a long-term predictor is modulated by both genetic and environmental factors. PMID- 10373232 TI - Intravascular source of adenosine during forearm ischemia in humans: implications for reactive hyperemia. AB - It is believed that adenosine is released in ischemic tissues and contributes to reactive hyperemia. We tested this hypothesis in the human forearm using microdialysis to estimate interstitial and intravascular levels of adenosine and caffeine withdrawal to potentiate endogenous adenosine and determine its effect on reactive hyperemia. Forearm blood flow response to ischemia was measured by air plethysmography before and 60 hours after the last dose of caffeine (250 mg TID for 7 days, n=6). Forearm blood flow increased by 274+/-66% and 467+/-97% after 3 minutes of forearm ischemia, before and during caffeine withdrawal, respectively (P<0.05). Thus, caffeine withdrawal enhances reactive hyperemia. To determine the source of adenosine, we measured interstitial adenosine with the use of a microdialysis probe inserted into the flexor digitorum superficialis muscle of the forearm, and we measured intravascular adenosine with the use of a microdialysis probe inserted retrogradely into the medial cubital vein. Dialysate samples were collected at 15-minute intervals during resting, forearm ischemia, and recovery periods. Forearm ischemia failed to increase muscle dialysate concentrations of adenosine but did increase intravascular dialysate adenosine 2.1-fold, from 0.61+/-0.12 to 1.28+/-0.39 micromol/L (P<0.01, n=8). Intravascular dialysate concentrations of thromboxane B2 did not increase during ischemia, ruling out platelet aggregation as a source of adenosine. These results support the hypothesis that endogenous adenosine contributes to reactive hyperemia and indicate that the major source of adenosine in the human forearm is intravascular. We speculate that endothelial cells are the source of intravascular adenosine during ischemia. PMID- 10373234 TI - Thermoregulatory and cardiac responses of infant spontaneously hypertensive and Wistar-Kyoto rats to cold exposure. AB - Cardiovascular function during cold exposure is dependent on effective thermoregulation. This dependence is particularly apparent in infants. For example, we have previously demonstrated that in infant rats during cold exposure, cardiac rate is directly related to their ability to produce heat endogenously. The primary source of endogenous heat production for infant rats is brown adipose tissue (BAT). Because of the dependence of cardiac rate on effective thermoregulation in the cold and because hypertension in spontaneously hypertensive rats (SHR) is influenced by the preweanling environment, in this study we examined the thermoregulatory and cardiac rate responses of infant SHR and Wistar-Kyoto rats (WKY) to varying levels of cold exposure. In experiment 1, 7- to 8-day-old SHR and WKY were acclimated at a thermoneutral air temperature (35 degrees C) and then exposed to successive decreases in ambient temperature (30.5 degrees C, 26.5 degrees C, 23 degrees C, and 17 degrees C) while thermal and metabolic measures were recorded. Although both strains increased BAT thermogenesis and oxygen consumption in response to cold exposure, SHR cooled more than WKY and exhibited lower levels of oxygen consumption at the lowest air temperatures. Experiment 2 was identical to experiment 1 except that cardiac rate was also measured. Again, SHR exhibited substantial thermoregulatory deficits compared with WKY; in addition, they were less able than WKY to maintain cardiac rate at the 2 lowest air temperatures tested. Finally, in experiment 3, infant SHR exhibited diminished BAT thermogenesis in response to a range of doses of a selective beta3-adrenoceptor agonist. We hypothesize that long-term thermoregulatory deficits during the early postnatal period influence cardiovascular function and contribute to the development of hypertension in SHR. PMID- 10373235 TI - Altered pressure-natriuresis in obese Zucker rats. AB - It has not been examined whether the pressure-natriuresis response is altered in the insulin-resistant condition. Furthermore, despite an important role of nitric oxide (NO) in modulating pressure-natriuresis, no investigations have been conducted assessing the renal interstitial NO production in insulin resistance. The present study examined whether pressure-natriuresis was altered in insulin resistant obese Zucker rats (OZ) and assessed the cortical and medullary nitrate/nitrite (NOx) levels with the use of the renal microdialysis technique. In OZ, serum insulin/glucose ratio (23.0+/-4.0x10(-8), n=9) and blood pressure (119+/-3 mm Hg) were greater than those in lean Zucker rats (LZ; 7.0+/-1.9x10(-8) and 103+/-4 mm Hg, n=9). The pressure-natriuresis curve in OZ was shifted to higher renal perfusion pressure (RPP), and the slope was blunted compared with that in LZ (0.073+/-0.015 vs 0.217+/-0.047 microEq/min kidney weight/mm Hg, P<0.05). The basal renal NOx level was reduced in OZ (cortex, 4.032+/-0.331 micromol/L; medulla, 4. 329+/-0.515 micromol/L) compared with that in LZ (cortex, 7.315+/-1. 102 micromol/L; medulla: 7.698+/-0.964 micromol/L). Furthermore, elevating RPP increased the medullary NOx in LZ, but this pressure-induced response was lost in OZ. Four-week treatment with troglitazone, an insulin sensitizing agent, improved hyperinsulinemia, systemic hypertension, and basal renal NOx levels (cortex, 5.639+/-0.286 micromol/L; medulla, 5.978+/-0.284 micromol/L), and partially ameliorated the pressure-natriuresis curves; the slope of pressure-natriuresis curves and elevated RPP-induced NOx, however, were not corrected. In conclusion, our study suggests that insulin resistance is closely associated with abnormal pressure-natriuresis and hypertension. These deranged renal responses to insulin resistance are most likely attributed to impaired medullary NO production within the medulla. PMID- 10373236 TI - Update on the systolic hypertension in Europe (Syst-Eur) trial. The Syst-Eur Investigators. PMID- 10373237 TI - Comparative behavioural effects of benzodiazepine and non-benzodiazepine anxiolytics in rhesus monkeys. AB - Quantitative behavioural assessment of benzodiazepines (chlordiazepoxide, diazepam and lorazepam) and non-benzodiazepines (buspirone) anxiolytics were investigated in unrestrained rhesus monkeys (Macaca mulatta) living in social colonies. The different behaviour, categorised as social, solitary and abnormal were video recorded and analysed. Chlordiazepoxide (2.5-5 mg kg-1, p.o.), diazepam (2.5-5 mg kg-1, p.o.) and lorazepam (0.5-1 mg kg-1, p.o.) induced dose dependent significant changes in certain social and solitary behavioural responses. Thus increases in social grooming, approach, contact, self grooming, feeding and resting with eyes open and decreased aggressiveness and vigilance. On the other hand buspirone (5-10 mg kg-1, p.o.) produced no significant alteration in social and solitary behavioural patterns. On the basis of the above findings the social and solitary behaviour protocol in non-human primates can be a useful tool for studying the effect of a new anxiolytic compound before clinical trial. PMID- 10373238 TI - Genetically modified mice in neuropharmacology. AB - Since the first transgenic mouse was reported in 1980, genetically engineered mice have become an invaluable biological tool for better understanding of physiological and pathological processes in many fields of biomedical research. The transgenic technology allows researchers to carry out specific genetic manipulation in all cells of a laboratory animal, and makes it possible to dissect gene function in a living organism. In the field of neurosciences these animals have contributed greatly to shed light on basic mechanisms of brain function as well as to generate useful animal models for studying human neurological disorders. In this review, the different techniques available for generating specific mutations in the mouse genome will be described, from pronuclear microinjection to gene targeting in embryonic stem cells, and to the second generation of inducible and conditional knockout mice. Then, the impact of transgenic mouse models as an alternative or additional approach to neuropharmacology will be discussed, not only for the study of molecular mechanisms in the central nervous system but also for the identification of new biological targets for innovative pharmacological therapy. PMID- 10373239 TI - Studies on the spasmolytic and uterine relaxant actions of n -ethyl and n -benzyl 1,2-diphenyl ethanolamines: elucidation of the mechanisms of action. AB - The influence of N -ethyl- and N -benzyl-1,2-diphenyl ethanolamines (compounds E and B, respectively) was examined on the spontaneously contracting rabbit jejunum and the rat uterus together with their influence on the contractions induced by some spasmogens in the guinea-pig ileum and oxytocics and CaCl2in the pregnant rat uterus. Both E and B inhibited the spontaneous contractions of the rabbit jejunum with ID50values of 0.13 and 0.03 micromol ml-1. Their inhibitory activities were not antagonized by alpha- or beta-adrenoceptor blockers but significantly reversed by CaCl2(0.015 micromol ml-1). The compounds also antagonized nicotine, ACh-, histamine-, 5-HT- and CaCl2-induced contractions by 44-100%. Compound E seemed to be several times more potent than B in inhibiting the spontaneous uterine contractions with an ID50of (7 nmol ml-1). Their inhibitory effects were not antagonized by beta2-adrenoceptor or H2-receptor blocking drugs. Both compounds (40 nmol ml-1) antagonized in a competitive manner CaCl2-induced contractions in the K+-depolarised uterus and PGE2and oxytocin induced uterine contractions. The ID50values were in the range of 1.6-10.7 nmol ml-1. The results suggest that E and B compounds may be considered as putative L Ca2+channel blockers with certain selectivities. The E compound seemed to be more selective against uterine L-Ca2+channels and the B compound against intestinal smooth muscles. Thus, the compounds may be of potential value in treatment of some colics, the irritant bowel syndrome, dysmenorrhoea and premature deliveries. PMID- 10373240 TI - The influence of lidocaine and verapamil on haemodynamic parameters after intravenous administration of midazolam in rabbits. AB - Cardiac arrhythmia can be a serious complication during general anaesthesia of patients suffering from cardiac arrhythmias and using antiarrhythmic drugs. The aim of the present experiments was to establish the way in which lidocaine and verapamil influence haemodynamic parameters of rabbits after midazolam anaesthesia. The experiments were performed on rabbits. Heart rate was counted according to ECG. Blood pressure was measured directly in the carotid artery. Cardiac output, stroke volume, and total peripheral resistance were estimated using the method of human 125J albumin dilution. Midazolam caused a gradual fall of arterial blood pressure till 30 min of the experiment. The decrease of blood pressure in the first 45 min of the experiment might be a result of decreasing peripheral resistance, and after 60 min should be rather attributed to a decrease of cardiac output and stroke volume. Lidocaine did not change the influence of midazolam on the blood pressure and heart rate. The consequence was a decrease of blood pressure after midazolam and lidocaine administration. Combined administration of midazolam and lidocaine decreased the influence of both midazolam and lidocaine on the total peripheral resistance. An injection of midazolam with verapamil resulted in a significant decrease of blood pressure. Combined administration of midazolam and verapamil caused a significant decrease of heart rate as compared with the initial value and administration of midazolam alone. PMID- 10373241 TI - Atenolol depresses post-ischaemic recovery in the isolated rat heart. AB - Metabolic events during ischaemia are probably important in determining post ischaemic myocardial recovery. The aim of this study was to assess the effects of the beta-blocker atenolol and the high energy demand in an ischaemia-reperfusion model free of neurohormonal and vascular factors. We exposed Langendorff-perfused isolated rat hearts to low-flow ischaemia (30 min) and reflow (20 min). Three groups of hearts were used: control hearts (n =11), hearts that were perfused with 2.5 micrograms l-1atenolol (n =9), and hearts electrically paced during ischaemia to distinguish the effect of heart rate from that of the drug (n =9). The hearts were freeze-clamped at the end of reflow to determine high-energy phosphates and their metabolites. During ischaemia, the pressure-rate product was 2.3+/-0.2, 5.2+/-1.1, and 3.3+/-0.3 mmHg 10(3)min in the control, atenolol and paced hearts, respectively. In addition, the ATP turnover rate, calculated from venous (lactate), oxygen uptake and flow, was higher in atenolol (11.2+/-1.7 micromol min-1) and paced (8.1+/-0.8 micromol min-1) hearts than in control (6.2+/-0.8 micromol min-1). At the end of reflow, the pressurexrate product recovered 75.1+/-6.4% of baseline in control vs 54.1+/-9.1 and 48.8+/-4.4% in atenolol and paced hearts (P<0.05). In addition, the tissue content of ATP was higher in the control hearts (15.8+/-1. 0 micromol g(dw)(-1)) than in atenolol (10.5+/-2.6 micromol g(dw)(-1)) and paced (10.9+/-1.3 micromol g(dw)(-1)) hearts. Thus, by suppressing the protective effects of down-regulation, both atenolol and pacing apparently depress myocardial recovery in this model. PMID- 10373242 TI - Effect of fluoxetine on maximal electroshock seizures in mice: acute vs chronic administration. AB - There are no definite reports regarding the effects of chronic fluoxetine on animal models of epilepsy. Since chronically administered fluoxetine, in comparison to acutely administered fluoxetine has different effects on CNS, the present study was undertaken to investigate the effect of acute and chronic fluoxetine pretreatment, on a median anticonvulsant dose (ED50) of phenytoin in male ICR albino mice. Additionally, the effects of fluoxetine pretreatment on median convulsive current (CC50) in the presence and absence of phenytoin were investigated and results were compared. The maximal electroshock seizure (MES) test was used to estimate the ED50of phenytoin. The electroshock threshold test was used to estimate CC50. ED50and CC50values were calculated by probit analysis. The effects of the chronic and acute fluoxetine groups on the ED50of phenytoin were significantly different (P<0.05), and on CC50this difference was not statistically significant. Chronic fluoxetine insignificantly increased the ED50of phenytoin and decreased the CC50while acute fluoxetine decreased the ED50of phenytoin and increased the CC50. Our results indicate that chronic fluoxetine does not have an antiepileptic property and it may have dubious proconvulsant properties, contrary to acute fluoxetine. PMID- 10373243 TI - Compartmental analysis revisited. AB - This paper gives a precise definition of compartment and shows that even when a compartmental model is not exactly identifiable, a range of its pharmacokinetic parameters can be determined. It is thus better to know approximate values of clearly defined parameters than exact values of quantities with no physical or physiological content. PMID- 10373244 TI - Neurochemical characteristics and behavioral responses to psychological stress in ovariectomized rats. AB - The present study was designed to clarify the time-dependent changes in brain monoamine turnover in the frontal cortex, hypothalamus, hippocampus, septum and amygdala after ovariectomy, and the difference in behavioral responses to psychological stress between sham-operated and ovariectomized (OVX) rats. At 2 and 4 weeks after ovariectomy, the turnover rates of dopamine and norepinephrine in all of the brain regions examined did not differ significantly between the sham-operated and OVX rats. However, 5-hydroxytryptamine (5-HT) turnover in all of the brain regions at 2 weeks after OVX was significantly lower than that in sham-operated rats. This difference was greater in the hypothalamus than in other brain regions. At 4 weeks after ovariectomy, 5-HT turnover in all of the brain regions examined was not significantly different between sham-operated and OVX rats. At 2 and 4 weeks after ovariectomy, exploratory behaviour (e.g., locomotor activity, head- dipping, crossing and rearing behaviours) in a non-stressed ovariectomy group did not differ from that in a non-stressed sham-operation group. Locomotor activity and the number of head-dips and crossings significantly (P<0.05) increased after repeated exposure to psychological stress for 5 days in sham-operated rats, but not in those at 2 weeks after OVX. At 4 weeks after ovariectomy, locomotor activity and the number of crossings and rearings in sham operated and OVX rats were not significantly different in the psychological stress and non-stress groups. However, the number of head-dips significantly (P<0.05) increased with psychological stress in the sham-operated rats, but not in OVX rats. These results suggest that female gonadal hormones may play an important role in the regulation of brain 5-HTergic systems. These interactions between gonadal hormones and 5-HT metabolism may be related to 5-HT-related neuropsychiatric disorders. PMID- 10373245 TI - Pharmacological analysis of 5-hydroxytryptamine effects on human isolated ureter. AB - 5-Hydroxytryptamine (5-HT) induced concentration-dependent contractions in human isolated ureteral strips in vivo. On the basis of available selective 5-HT agonists and antagonists, we have further investigated the receptors involved. At concentrations from 10 n m to 1 m m, 5-HT induced concentration-dependent contractions. Significant contractions were not observed with 5-HT1Aagonist 8-OH DPAT (10(-9)-10(-4)m), 5-HT1Dalphaagonist sumatriptan (10(-9)-10(-4)m), 5 HT2agonist DOI (10(-9)-10(-4)m), 5-HT3agonist 2-methyl 5-HT (10(-9)-10(-3)m) and 5-HT4agonist renzapride (10(-9)-10(-3)m) on the human isolated ureter. On the other side, a 5-HT1-likeagonist 5-CT (10(-9)-10(-3)m) produced contractions on the isolated samples. The Emaxdeveloped by 5-CT was significantly smaller than that of the 5-HT (29% of 5-HT). Methithepin, the less selective 5-HT1/2antagonist (10(-9)-10(-6)m), 5-HT3antagonist, ondansetron (10(-9)-10(-5)m) and 5 HT4antagonist DAU 6285 (10(-8)-10(-6)m) did not antagonise the contractile responses to 5-HT. 10(-7)m ketanserin antagonised 5-HT induced contractile responses in ureteral strips. Additionally, combined administration of 5 HT4antagonist DAU 6285 (10(-6)m) and 5-HT1/2antagonist methithepin (10(-6)m) caused a rightward shift of the CRC of 5-HT yielding pEC50values of 4.68+/-0.15. 5-HT-induced contractile responses that were not abolished by TTX and atropine, thus supporting the suggestion that in the human, the contractile responses to cumulative addition of 5-HT of the ureter are not mediated by excitation of cholinergic neurons. In the present study the receptor mediating the contractile response to 5-HT in the human upper ureter could not be clearly designated 5-HT1 like, 5-HT2, 5-HT3or 5-HT4. This study suggests that contractile response to 5-HT in the upper segments of the human ureter appear to be mediated by an atypical 5 HT receptor subtype. PMID- 10373246 TI - [3H]Tyramine uptake by rat liver slices. AB - Rat liver slices were employed as experimental model to characterise the system involved in the transport process which participates in liver tyramine uptake. The uptake of 0.4 micromol l-1of [3H]tyramine by rat liver slices was linear from 5 min up to the end of incubation. At 15 min the uptake was 4.58+/-0.18 pmol mg 1protein. The accumulation of [3H]tyramine was sensitive to temperature (69. 3+/ 4.0% inhibition at 0 degrees C, P<0.001), to sodium omission replaced by 150 mmol l-1Tris or 110 mmol l-1Tris+40 mmol l-1choline (27.6+/-6.0%, P<0.01, and 24.6+/ 3.8% inhibition, P<0.01, respectively), and the inhibition of Na+-K+-adenosine triphosphatase by 150 micromol l-1ouabain (20.4+/-2.6% decrease, P<0.01). Uptake of [3H]tyramine was cocaine- (10 micromol l-1) and desipramine- (1 micromol l-1) dependent (32.2+/-6.4%, P<0.05, and 31.6+/-4.0% inhibition, P<0.05, respectively). Uptake of [3H]tyramine in rat liver slices was not modified by 30 micromol l-1isoprenaline, 30 micromol l-1corticosterone, 30 micromol l 1normetanephrine and noradrenaline up to 4 micrometers at higher noradrenaline concentrations tyramine transport was diminished (P<0.05). Results achieved by incubation with increasing tyramine concentrations indicate that at the micromolar level hepatic uptake occurs by a combined passive diffusion and transport-mediated mechanism, whereas at greater tyramine concentrations passive transport predominates. These results suggest that both simple diffusion and a transport-mediated mechanism are involved in this uptake from hepatocytes, which presents features similar to those described for type 1 non-neuronal uptake systems. PMID- 10373247 TI - Dopaminergic projection to the central amygdala mediates the facilitatory effect of CCK-8US and caerulein on memory in rats. AB - The involvement of dopaminergic projection to the central amygdala in the facilitatory effect of cholecystokinin unsulphated octapeptide (CCK-8US) and caerulein (CER) on memory motivated affectively was investigated in male rats. CCK-8US and CER were administered subcutaneously at the doses of 10 micrograms kg 1and 0.5 microgram kg-1, respectively, immediately after a single learning trial in a passive avoidance situation, after bilateral 6-OHDA lesions to the central amygdala. Bilateral 6-OHDA lesions to the central amygdala totally abolished the facilitatory effect of CCK-8US and CER on retention of passive avoidance behaviour evaluated 24 h after the learning trial. These results may indicate that the facilitatory effect of CCK-8US and CER on memory motivated affectively is mediated by dopaminergic projection from ventral tegmental area to the central amygdala. PMID- 10373248 TI - Genetic models in pharmacology: present status and future. PMID- 10373249 TI - Finding a parent in a king penguin colony: the acoustic system of individual recognition. AB - To be fed, a king penguin, Aptenodytes patagonicus, chick must identify the call of its parents, in the continuous background noise of the colony. To study this recognition process, we played back to the chicks parental calls with acoustic parameters modified in the temporal and frequency domains. The parental call is composed of syllables (complex sounds with harmonic series) separated by pronounced amplitude declines. Our experiments with modified signals indicate that the chick's frequency analysis of the call is not tuned towards precise peak energy values, the signal being recognized even when the carrier frequency was shifted 100 Hz down or 75 Hz up. To recognize the adult, chicks used frequency rather than amplitude modulation, in particular the frequency modulation shape of the syllable. This structure is repeated through the different syllables of the call giving a distinct vocal signature. Our experiments also show that the receiver needs to perceive only a small part of the signal: the first half of the syllable (0.23 s) and the first three harmonics were sufficient to elicit recognition. The small amount of information necessary to understand the message, the high redundancy in the time and frequency domains and the almost infinite possibilities of coding provided by the frequency modulation signature permit the chick to recognize the adult, without the help of a nest site. For these reasons, the code used in the call of the king penguin can be regarded as a functional code, increasing the possibility of individual recognition in an acoustically constraining environment. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10373250 TI - Long-term memory for a spatial task in young chicks. AB - Two-day-old chicks, Gallus gallus domesticus, were faced with a spatial task requiring them to make a detour around a U-shaped barrier in order to join a group of conspecifics placed beyond it. Chicks made one detour trial, and were then retested after delays of 30 min, 3 h and 24 h. When retested, the chicks took significantly less time to make the detour, even at 24 h. Chicks that failed to solve the task on the first trial within the time limit (600 s) took as long as naive chicks, when tested again 24 h later, suggesting that long-term memory for the task requires a form of one-trial learning. Since many chicks chose the same direction of detour on both the first trial and the retest, they may have simply shown a stereotyped preferential response. In a further experiment to test this possibility, we used a more complex version of the apparatus: the direction of detour first chosen by the chick was always blocked, and a subsequent detour in the opposite direction was required to complete the task. This time, chicks did not choose the same direction on the first test and when tested 24 h later. However, they proved able to remember the correct direction for at least 24 h if they were allowed to learn the correct path in a series of five trials. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10373251 TI - Avoidance of superparasitism: a matter of learning? AB - Superparasitism (laying eggs into parasitized hosts) by solitary parasitoids was regarded for a long time as a mistake on the part of the foraging parasitoid, but is now widely accepted as often adaptive. In Venturia canescens the rate of avoidance of superparasitism has been shown to rise over the first 20 min from the deposition of the first egg, possibly because of a constraint in the detectability of the marker used to label parasitized hosts. Here, we show that the increase in avoidance of superparasitism with time is the result of a female's experience of hosts in the interval between laying an egg in a host and re-encountering that same host. Wasps deprived of hosts in this interval showed no avoidance of superparasitism; those given healthy hosts every 3 min during this interval showed increasing avoidance of superparasitism with time. Furthermore, the marker was detectable in a host within 3 min of oviposition. The results suggest that wasps quickly acquire information about the abundance of healthy hosts in their environment, and base their decision to superparasitize on this information. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10373252 TI - Parasites promote mating success: the case of a midge and a mite. AB - I tested the prediction that the hydracharinid mite Unionicola ypsilophora reduces mating success in the chironomid midge Paratrichocladius rufiventris. Males of the midge form mating swarms through which females fly to emerge after a few minutes with a mate. This mating system is believed to depend upon the male capturing a mate after aerial competition between males. Thus aerobatic ability is expected to determine success and a large ectoparasite should impair aerial performance. The proportion of infested males in swarms (ca. 4%) was less than that in mated pairs (ca. 15%). Infestation thus improved the expected mating success of the male midge. This finding is counterintuitive and may be a chance effect of no adaptive value to host or parasite. Alternatively, it may be an adaptive manipulation of the host by the parasite. This study provides evidence for the latter explanation. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10373253 TI - Reproductive behavioural sequences of single pairs of Atlantic salmon in an experimental stream. AB - We studied 12 size-matched pairs of Atlantic salmon, Salmo salar, in an experimental stream in southwest France, to determine whether fish activity and motivation changed during the course of reproduction. The absolute weight of spawners did not affect their spawning activity. On average, females deposited their eggs within 3 days in nine nests. Male and female breeding behaviours changed throughout the reproductive period. This cyclic variation in behaviour appeared to be determined in part by the activity of the other sex, as a consequence of complex interplay between the sexes, but also largely by the stage of the spawning period. During the first three ovipositions, male-female stimulus reaction chaining became more consistent just before spawning, which may help synchronize gamete release for successful fertilization. During the last three ovipositions, sequence chaining between the sexes was less coherent, possibly as a result of reduced mate attractiveness and/or physiological limitations. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10373254 TI - Temporal, spatial and social segregation of red-billed choughs between two types of communal roost: a role for mating and territory acquisition. AB - We assessed the value of communal roosting for mating and territory acquisition in nonbreeding red-billed choughs, Pyrrhocorax pyrrhocorax. Between 1991 and 1996 we surveyed all communal roosts, breeding territories and vacant nest sites in an area of 250 km2in Los Monegros (northeast Spain), as well as other communal roosts in the surrounding 500 km2from 1987 to 1996. Main roosts were used traditionally and throughout the year and gathered more choughs than subroosts, which were used only during the nonbreeding season. Nonbreeding choughs were socially segregated in these roosts according to their age and breeding prospects. Compared with random potential roost sites, main roosts included more first-year birds and were in areas of higher breeding density. Subroosts were mainly joined by choughs of breeding age (at least 2 years old) whose morphology (males) and body condition (females) were similar to those of established breeders, and were in areas with breeding densities as high as around main roosts but with more available nest sites. Nonbreeding choughs that used main roosts later used subroosts, while the converse was rare. More choughs joining subroosts acquired mates than those joining main roosts. Choughs mostly paired with roostmates but also with widowed territorial breeders. Finally, choughs from subroosts acquired territories closer to the roost than those who mated in main roosts, supporting the hypothesis that subroosts reduce the costs of mating and territory acquisition. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10373255 TI - Male choice for female colour morphs in Ischnura elegans (Odonata, Coenagrionidae): testing the hypotheses. AB - The occurrence of different conspecific female colour morphs, with one of the morphs resembling the male, is supposed to have consequences for mate choice. There are two hypotheses linking mate choice and female colour polymorphism. First, males may mate predominantly with female morphs that differ from the male because they do not recognize androchrome females as females (male mimic hypothesis). Second, males may be more attracted to the most common morph in the population (habituation hypothesis). We tested these hypotheses in five populations of the same species, Ischnura elegans, with a range of androchrome frequencies. In each population we performed binary choice experiments in small cages. Males did not consistently prefer gynochrome females but mated predominantly with the most common morph in the population. Moreover, a reanalysis of the available damselfly data in the literature also supported the habituation hypothesis. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10373256 TI - Stripes display in hover-wasps (Vespidae: Stenogastrinae): a socially costly status badge. AB - During their daily patrols of their hover sites, male stenogastrine wasps display three white stripes on their tergites by fully stretching their abdomen. In captive Parischnogaster mellyi males, we observed a positive relationship between mating and both display frequency and successful aerial duels. Approaches of receptive females to hovering males and sexual interactions were most frequent at the end of the males' performance, when only a few individuals displayed their stripes in flight. We investigated the function and cost of the stripes display by manipulating this sex-dimorphic trait. Wasps with additional white stripes (simulating continuously displaying individuals) were pursued and touched more frequently by rivals, stopped their activity earlier than controls and foraged more intensely. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10373257 TI - Effect of experience on predatory behaviour of dogwhelks. AB - We used an acoustic transducer to monitor the radular activity of dogwhelks, Nucella lapillus, drilling mussels, Mytilus edulis, in the laboratory and we examined the effect of dietary experience on prey-handling behaviour. For the first time, phases of inspection, penetration and ingestion could be distinguished directly, and consequently the prey-handling process analysed in detail. Dogwhelks with different field-based experience of mussels showed different handling behaviour. Those collected from a mussel-dominated shore more readily adopted the faster method of penetrating between the slightly gaping valves, instead of the slower method of drilling through the shell. Those collected from a barnacle-dominated shore took significantly longer to attack the mussel and then were unable to switch from drilling to penetrating through the gape between valves. Experience of specific prey in the field, by reducing handling time, could promote fitness by reducing exposure to environmental hazards. Laboratory attempts to train dogwhelks from the barnacle-dominated shore to use the gape penetration method failed, suggesting that functional constraints, such as injection of a relaxant when penetrating through the gape and/or genetically controlled behavioural traits, could limit the ability to learn handling skills. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10373258 TI - Feeding experience affects web relocation and investment in web threads in an orb web spider, Cyclosa argenteoalba. AB - Orb-web spiders often relocate their webs when they assess a web site as prey poor. When a spider spins its web at a new site, it may not be able to assess the prey availability at the site accurately on the first day, owing to stochastic variation in foraging success, but it is gradually able to make an assessment. Therefore, a spider's foraging behaviour may change according to how long it has been at its current web site. To test this possibility, we conducted a prey removal experiment, with the spider Cyclosa argenteoalba, to compare the response to prey deprivation of spiders that were on new sites with that of spiders that had been at a site for several days. Spiders in both groups had a higher relocation rate than the natural rate, but more spiders in the new-site group relocated their webs. Spiders thus seemed to use previous experience of prey capture at a web site to decide whether to relocate their web. The total length of silk thread in a web was greater on the second day at a new web site than on the first. We suggest that spiders minimize their investment in web threads until they are certain that the web site is prey rich. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10373259 TI - The development of within-song type variation in song sparrows. AB - We investigated the development of within-song type variation in song sparrows, Melospiza melodia, with two experiments designed to determine how exposure to within-type variation influences the song-learning process and whether within type variation itself is a learned trait. In the first experiment, we compared learning between two groups of males, one group tutored exclusively with song models presented with no variation, and the other group tutored exclusively with song models presented with a range of within-type variation that is normally produced by birds in the field. The two groups in this experiment did not differ significantly in any measure of how well they learned, suggesting that exposure to within-type variation has no measurable influence on the learning process overall. Nor did the groups differ in the expression of within-song type variation in their own adult songs, demonstrating that within-type variation is not a learned feature of song sparrow song. In the second experiment, we tutored a single group of birds with both invariant and variable models, allowing us to ask how within-type variability affects learning preferences. Young birds preferentially copied song type models presented with variation significantly more than invariant models. Taken together, these experiments provide insight into the evolution of within-song type variation in song sparrows, although the functional significance of this level of variation and learning preferences based on variation remain enigmatic. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10373260 TI - Signal-receiver interplay in the communication of male condition by Asian elephants. AB - Signal design and meaning are dependent on the condition of the sender and receiver as well as the response of the receiver. This study examined (1) whether female Asian elephants, Elephas maximus, can distinguish between a conspecific male in musth and nonmusth states using urinary signals, (2) how the oestrous condition of the female affects discrimination, and (3) correlation of female responses with the testosterone level of the male. Musth is a rut-like state displayed by healthy adult male elephants. Males in musth dominate nonmusth males and may be preferred by females as mates. Urine was collected from two captive male Asian elephants during nonmusth periods and from one of these males during times of musth. Samples of musth and nonmusth urine and control liquids were placed in an elephant enclosure weekly for 16 weeks, the length of a female oestrous cycle. Primary response behaviours were approach and four trunk-tip motions, namely sniff, check, place and flehmen. Musth urine consistently elicited greater responses than nonmusth and control samples. Females were more responsive during their follicular (sexually receptive) than luteal (unreceptive) stages of oestrus. Furthermore, females appeared to be sensitive to the degree of musth as responses increased with rising serum testosterone levels of the male donor. Chemical signals from males are a likely source of honest signals related to status and reproductive condition. Female elephants appear capable of detecting differences in a male based upon urinary chemosignals. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10373261 TI - Song development by chipping sparrows and field sparrows. AB - When, where and from whom young songbirds learn their songs have been controversial issues in the study of song development. We chose to study some of these issues in two migratory and closely related songbirds, the chipping sparrow, Spizella passerina, and field sparrow, Spizella pusilla. Nestlings of both species were collected in western Massachusetts and hand-reared in the laboratory. There, juveniles were placed in separate cages and assigned to one of three rooms; in each room were eight young chipping sparrows, eight young field sparrows and two adult tutors of each species, arranged so that most of the young males were adjacent to adult tutors of the same species. During mid-winter, adult tutors were moved from one room to another, so that the young birds heard different song types from different tutors during their hatching year and the following spring. From spectral analysis of our extensive tape recordings, we found that most juvenile males imitated the songs of their hatching-year tutors but then gradually modified their songs to match more closely either their adult tutors or other pupils the next spring. One chipping and one field sparrow clearly imitated a new song syllable from a spring live tutor; that is, these yearling males learned songs by 'instruction'. Other sparrows improvised extensively, and one chipping sparrow learned a field sparrow's song syllable. Our results reveal great individual variation in how songs are developed, and we expect similar flexibility among birds in nature. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10373262 TI - Demographic consequences of incest avoidance in the cooperatively breeding acorn woodpecker. AB - The avoidance of breeding with close relatives is an adaptation to inbreeding depression. Unfortunately, inbreeding depression has proved difficult to document or measure in the wild, despite being frequently observed among animals in captivity. We address this problem indirectly by determining the demographic cost of incest avoidance in the cooperatively breeding, polygynandrous acorn woodpecker, Melanerpes formicivorus, following the death or disappearance of all breeders of one sex within a group (a reproductive vacancy). Groups undergoing female vacancies that also contained female nonbreeding helpers experienced significantly lower reproductive success in each of the subsequent 3 years than those in which either no nonbreeding helpers or only male nonbreeding helpers were present, a decrease attributable to incest avoidance between the helper females and the related breeder males in the group. Using a computer simulation combined with a life-table analysis, we estimated that incest avoidance costs the population 9.2-12.1% in overall reproductive potential (measured in fledglings/female) and decreases the population rate of increase by 1.78 2.33%/year. These results suggest the presence, on average, of at least 1.2-1.8 lethal equivalents per individual, a value of the same magnitude as estimated for several other taxa, including humans. Incest avoidance may compound random demographic and environmental events and significantly facilitate the decline of threatened populations even prior to any detrimental effects of inbreeding depression per se. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10373263 TI - Identification of a disturbance signal in larval red-legged frogs, Rana aurora. AB - Animals that are warned about the presence of a predator are more likely to avoid and/or survive an encounter with a predator. Chemical signals released by disturbed or injured conspecifics may provide prey animals with an early warning. In this study we conducted experiments to determine whether larval red-legged frogs respond to chemical stimuli produced by disturbed conspecifics and to examine the chemical compounds that may act as the alarm signal. In laboratory tests, groups of tadpoles responded with antipredator behaviours when exposed to chemical cues of disturbed conspecifics but not when exposed to chemical cues of control (undisturbed) conspecifics. In subsequent tests, disturbed animals increased ammonium (the main metabolic waste of tadpoles) excretion relative to undisturbed individuals. When tadpoles were exposed to low-level ammonium concentrations (1 mg NH4+/litre), they responded by increasing antipredator behaviours. Our results suggest that red-legged frog tadpoles release a chemical that provides conspecifics with an early warning of predator presence, and that ammonium (NH4+) may be a component of the disturbance signal. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10373264 TI - Assessment of rattlesnake dangerousness by California ground squirrels: exploitation of cues from rattling sounds. AB - We propose that the predator-prey relationship between California ground squirrels, Spermophilus beecheyi beecheyi, and northern Pacific rattlesnakes, Crotalus viridis oreganus, offers a compelling analogy with the well-studied case of intraspecific fighting and assessment. Because ground squirrels frequently place themselves at risk by harassing rattlesnakes, they stand to benefit from assessment strategies which serve to mediate risk. For example, larger and warmer snakes are more dangerous than smaller and cooler ones. These determinants of dangerousness covary with acoustic characteristics of the rattling sounds elicited by squirrel harassment. To determine whether squirrels use these acoustic cues regarding rattlesnake body size and body temperature, we played back rattling and control sounds to individuals in a population of free-living squirrels. The squirrels clearly associated rattling sounds with rattlesnakes and proved capable of assessing both determinants of rattlesnake dangerousness on the basis of acoustic cues. Several features of squirrel behaviour covaried with these acoustic cues, including tail flagging, bipedal posture, and hesitancy to reapproach the area where the rattle was heard. Many of these behavioural differences were sustained for up to 10 min postplayback. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10373265 TI - Black-capped chickadee call dialects along a continuous habitat corridor. AB - The gargle call, a vocalization used in agonistic encounters by black-capped chickadees, Poecile atricapillus, was examined for evidence of geographical variation along a corridor of continuous riparian habitat in northern Colorado. We captured birds from three different sites during the nonbreeding season and brought them into the laboratory, where their gargle calls were recorded. We sorted sonagrams produced from these vocalizations visually into distinct gargle types having similar compositions of individual units, or syllables. This allowed us to characterize both individual and population repertoires. The majority (88.7%) of gargle types analysed were found to be unique to individual populations rather than shared among or between populations. Examination of individual repertoires showed that chickadees shared a higher proportion of gargle types with birds from their own sites compared with birds from either of the two other sites. Thus, gargle dialects occurred among these chickadee populations despite the absence of geographical barriers to blending of vocal traditions. As the birds studied were obtained from sites along an uninterrupted dispersal corridor, the results of this study suggest that behavioural mechanisms are responsible for maintenance of dialects in this aggressive call. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10373266 TI - Trophallaxis in the honeybee, Apis mellifera : the interaction between viscosity and sucrose concentration of the transferred solution. AB - Trophallaxis by honeybee foragers was studied under the experimental conditions of an arena. The behaviour of pairs of bees, one (donor) fed with 50-&mgr;l sucrose solutions and another unfed recipient, was analysed as a function of the sucrose concentration, the concentration at constant viscosity (kept constant by adding tylose, an inert polysaccharide), and of the viscosity of a 30% sucrose solution. By increasing the concentration of solutions, the rate at which the solution was transferred to recipient bees (transfer rate of solution, in &mgr;l/s) increased up to a maximum value for 30% sucrose solution, and decreased beyond this concentration (concentration experiment). At constant viscosity, no modulation was observed for the lower sugar concentration range (10-30%), while the transfer rate of solution clearly increased beyond 30% (concentration experiment at constant viscosity). For the 30% sucrose solution, the transfer rate decreased with increasing viscosity (viscosity experiment). If only the sucrose compound is comparatively analysed, the transfer rate of sucrose (in mg/s) increased similarly in the first two experiments. These results give behavioural evidence suggesting that donor bees are capable of modulating the trophallactic food transfer as related to the sucrose concentrations carried into their crops within a considerable wide range, but viscosity prevents it. It also suggests that trophallactic transfer rate does not depend on abdominal volume, for even when all donor bees attained similar loads (50 &mgr;l), transfer rate of solution increased along with the offered sucrose concentration. Results are discussed in relation to the information exchange performed in the foraging context displayed by foragers. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10373267 TI - Patterns of movement and orientation during caching and recovery by Clark's nutcrackers, Nucifraga columbiana. AB - Clark's nutcrackers regularly store large numbers of pine seeds and remember the locations of the cached seeds. Although they are very accurate, they do make some errors during recovery. In an attempt to determine whether any behaviours during caching predicted the occurrence of errors during recovery, we videotaped Clark's nutcrackers while they cached and recovered seeds under laboratory conditions. We used the videotapes to develop complete, quantitative descriptions of caching and recovery behaviour, with an emphasis on body orientation and directions of movement. During caching, the birds showed the greatest change in their orientation and direction following cache creation. During cache recovery, in contrast, body orientation changed most following successful recovery of a seed. When orientation while making a cache was compared with orientation when recovering the same cache, orientations were similar more often than would be expected by chance. However, this consistency of direction was not related to the accuracy of cache recovery, indicating that such consistency is not necessary for accurate cache recovery. The location in which the birds chose to place their caches was the only variable that predicted the location of probes during recovery. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10373268 TI - Female choice and plasticity of male calling behaviour in the Pacific treefrog. AB - Male Pacific treefrogs, Hyla regilla, use advertisement and encounter calls to regulate intermale spacing within breeding choruses. When either type of call produced by a neighbour is detected above a particular amplitude, a resident frog responds aggressively by producing encounter calls. These 'aggressive thresholds' differ for the two call types and are plastic: males rapidly resume advertisement calling (accommodate) following repeated presentation of these calls above their aggressive threshold. We tested the hypothesis that this plasticity is the result of female preference for the advertisement call over the encounter call. Female choice was tested in a two-speaker phonotaxis assay with alternating presentation of the two call types. Of the 12 females that met our criteria for demonstrating a phonotaxic response, each approached the speaker playing the advertisement call rather than the encounter call. This strong female preference for the advertisement call supports our hypothesis, and suggests that the plasticity in male calling behaviour allows males to maximize the time spent in producing advertisement calls to attract a female by rapidly adjusting their aggressive behaviour to changes in male spacing within choruses. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10373269 TI - Organizing and archiving private collections of tape recordings. PMID- 10373271 TI - Contents of Volume 57. PMID- 10373270 TI - Testing association patterns of social animals. PMID- 10373272 TI - Introduction beyond stress: the role of individual difference factors in psychoneuroimmunology. PMID- 10373274 TI - Worry affects the immune response to phobic fear. AB - Worry, the cognitive enumeration and anticipation of potential future negative events, is associated with autonomic dysregulation, which may in turn have implications for the immune system. People endorsing high (n = 7) and normal levels of trait worry (n = 8) were briefly exposed to a phobic stimulus and the autonomic and immune responses and recovery were assessed. A time-matched control group (n = 6) was not exposed to any stimulus. Both worry groups showed increased heart rate and skin conductance in response to phobic fear. However, only the normal worry group showed a concomitant increase in natural killer cells in peripheral blood. Patterns of change during the follow-up period suggested that phobic fear had disrupted a normal circadian increase in natural killer cells. Adrenergic and hypothalamus-pituitary-adrenal mechanisms may be responsible for the differences between high and normal worry groups in their natural killer cell response to and recovery from phobic fear. PMID- 10373273 TI - Individual variability and immunity. PMID- 10373275 TI - Individual differences in prefrontal activation asymmetry predict natural killer cell activity at rest and in response to challenge. AB - Reliable individual differences in electrophysiological measures of prefrontal activation asymmetry exist and predict dispositional mood and other psychological and biological indices of affective style. Subjects with greater relative right sided activation report more dispositional negative affect and react with greater intensity to negative emotional challenges than their left-activated counterparts. We previously established that such individual differences in measures of prefrontal activation asymmetry were related to basal NK function, with left-activated subjects exhibiting higher levels of NK function than right activated subjects. The present study was designed to replicate and extend these earlier findings. Subjects were tested in five experimental sessions over the course of 1 year. During the first two sessions, baseline measures of brain electrical activity were obtained to derive indices of asymmetric activation. During sessions 3 and 4, blood samples were taken during a nonstressful period in the semester and then 24 h prior to the subjects' most important final examination. During session 5, subjects were presented with positive and negative film clips 30 min in duration. Blood samples were obtained before and after the film clips. Subjects with greater relative right-sided activation at baseline showed lower levels of basal NK function. They also showed a greater decrease in NK function during the final exam period compared to the baseline period. Subjects with greater relative left-sided activation showed a larger increase in NK function from before to after the positive film clip. These findings indicate that individual differences in electrophysiological measures of asymmetric prefrontal activation account for a significant portion of variance in both basal levels of, and change in NK function. PMID- 10373276 TI - Personality and tonic cardiovascular, neuroendocrine, and immune parameters. AB - Although there is now abundant evidence that certain personality features constitute risk factors for negative health outcomes, personality measures have received little attention to date in the behavioral immunology literature. The present study assessed the relationship between major dimensions of personality and tonic cardiovascular, neuroendocrine, and immunologic parameters in 276 healthy adults. Participants who scored low in agreeableness tended to have higher levels of systolic blood pressure, diastolic blood pressure, and epinephrine. Low levels of extraversion were associated with higher blood pressure, epinephrine, norepinephrine, and natural killer cell cytotoxicity. Neuroticism was generally unrelated to physiologic outcomes. Personality was not associated with leukocyte subset counts. The magnitude of relationships between personality and physiology was modest, with personality measures accounting for 1 to 7% of the variance in selected physiological parameters. Health practices did not mediate associations between personality and physiologic outcomes. However, a substantial proportion of the relationship between extraversion and natural killer cell cytotoxicity was accounted for by their common association with epinephrine and to a lesser extent norepinephrine. These findings are consistent with the notion that personality contributes to basal physiology and provide a foundation for further research on the relationship between personality and natural killer cell cytotoxicity. PMID- 10373277 TI - A preliminary description of responses of free-ranging rhesus monkeys to brief capture experiences: behavior, endocrine, immune, and health relationships. AB - A cohort of free-ranging rhesus monkeys has been followed since birth in 1994 on the island of Cayo Santiago, Puerto Rico. At 3 years of age, subjects were trapped and blood samples were collected after capture and prior to release the following day. Blood samples were processed for natural cytotoxicity toward xenogeneic tumors, phenotyping, and plasma hormones. Intestinal parasites were determined from fresh stool samples collected during trapping. Data were also available from the previous year for antibody titers to latent viruses prevalent in this population. Behavioral traits of each monkey were characterized using a previously developed trait scale for rhesus monkeys. Natural cytotoxicity toward both K562 and Raji targets declined from capture until release the following day. Plasma cortisol rose and plasma prolactin and growth hormone fell during the period of captivity; a rise in insulin was significant. It was expected that individual differences in behavioral traits might predict immune and hormone levels at the time of capture or changes in these parameters during the capture period. Although behavioral adjectives tended to cluster along three orthogonal dimensions (Insecurity, Irritability, and Sociability), they bore no relationship to the physiological parameters collected acutely (in vitro immune and endocrine parameters). The individual difference markers of gender and maternal rank were not related to the magnitude of the observed changes in these in vitro parameters, either. However, an in vivo measure (CMV titer) was related to individual differences in Irritability. It was concluded that the magnitude of the stress associated with capture overwhelmed the individual difference effects. PMID- 10373278 TI - The relationship of personality dimensions in adult male rhesus macaques to progression of simian immunodeficiency virus disease. AB - Studies of nonhuman primate personality have suggested that physiological correlates of relevant behavioral dimensions exist. The present study examined personality using techniques similar to those employed in human personality research. Adult male rhesus monkeys were each rated on 25 adjectives while living in their natal groups. Approximately 1.5 years later, 18 animals were inoculated with the simian immunodeficiency virus (SIV) and exposed to socially stable or socially unstable conditions. Behavior, viral load (SIV RNA), plasma cortisol concentrations, and the IgG response to SIV and to rhesus cytomegalovirus were measured at regular intervals. Multiple regression analyses revealed that the four personality dimensions (Sociability, Confidence, Equability, Excitability) were correlated with various measures. Following inoculation with SIV, animals higher in Sociability showed a more rapid decline in plasma cortisol concentrations, elevations in the anti-RhCMV IgG response, and a decline in SIV RNA. The results indicate that personality factors in rhesus monkeys do have physiological correlates that have significance for disease processes and that in the context of a social manipulation, Sociability, reflecting the tendency to engage in affiliative interactions, is an important factor in explaining outcome measures at early time points. PMID- 10373279 TI - Differential immune system changes with acute and persistent stress for optimists vs pessimists. AB - This study investigated whether acute and persistent stressors and life change events were followed by changes in immune status, and whether dispositional optimism moderated these relationships. Thirty-nine healthy women ages 18-45 were followed prospectively for 3 months, with weekly assessment of acute and persistent stressors and monthly assessment of life events and immune parameters (NK cell cytotoxicity, and CD4 and CD8 T cell subsets). The study used an autoregressive linear model to examine how weekly appraised acute and persistent stress levels were associated with immune parameters in the subsequent week. Analyses revealed that the immune outcomes were differentially affected by acute and persistent stressors. Further, the association between acute stress and subsequent immune parameters was buffered by an optimistic perspective. However, when stress persisted at high levels, optimists showed more subsequent immune decrements than pessimists. PMID- 10373280 TI - Differences in NK cell function in mice bred for high and low aggression: genetic linkage between complex behavioral and immunological traits? AB - In previous studies, we found differences in cellular immune responsiveness in Institute for Cancer Research (ICR) mice selectively bred for high and low levels of aggression. Compared to the high aggressive line, the low aggressive line had low levels of natural killer (NK) and T cell activity and increased susceptibility to tumor development. To dissect further this novel association, experiments were designed to test two competing hypotheses. The first hypothesis was that the phenotypic expression of the line differences in NK cell activity are dependent on and regulated by the expression of high and low levels aggressive behavior in the lines. The alternative hypothesis was that the differences in immune status are independent of the expression of aggression by the lines, suggesting linkage between a subset of genes involved in determining these complex behavioral and immunological traits or pleiotropic gene effects on both traits. In Experiment 1, three conditions of postweaning social experience (mice singly housed, group housed within line, or group housed between lines) were tested in males to determine whether experiential conditions which modify the expression of aggression would in turn modify the line differences in NK cell activity. This experiment revealed that the difference in NK cell activity between high aggressive and low aggressive male mice was attributable to line only. The different postweaning social conditions examined had no effect on modifying the differences in NK activity, and social dominance hierarchy did not correlate with levels of NK cell activity. Whereas males of the high and low lines exhibit differences in aggressive behaviors across most contexts, females do not exhibit such differences except in response to an intruder during the postpartum period. Therefore, in Experiment 2 we compared the NK cell activity of nulliparous females of the high and low aggressive lines. Under these conditions, females of the low aggressive line had low levels of NK activity compared to high aggressive females (differences comparable to those seen between males of the high and low lines). Taken together, these experiments lend support to the hypothesis that this association may be due to a genetic linkage between subsets of genes involved in determining these complex behavioral and immunological traits, or may possibly represent a fortuitous association which occurred during the selective breeding. PMID- 10373281 TI - Socially inhibited individuals show heightened DTH response during intense social engagement. AB - To determine whether altered cellular immune response might mediate the increased health risks associated with social inhibition, we examined delayed type hypersensitivity (DTH) responses in 36 adults under conditions of low and high intensity social engagement. Participants come from a study of psychological factors in functional bowel disease and fibromyalgia. Under high engagement conditions, socially inhibited individuals showed significantly increased induration in response to intradermal tetanus toxoid. Under low engagement conditions, these individuals showed less pronounced DTH responses that did not differ in magnitude from those of uninhibited individuals. This pattern of results was found using two different measures of social inhibition and was independent of social inhibition's definition as a continuously distributed trait vs a discrete category. These data are consistent with the general hypothesis that social inhibition represents a predisposition to physiologic hyperresponsiveness that requires an exogenous social trigger for expression. PMID- 10373283 TI - Abstracts PMID- 10373284 TI - Refereed papers PMID- 10373285 TI - Shape-Set dimensionality vs structural-distance effects in a patient with category-specific visual agnosia PMID- 10373286 TI - Semantic proximity and shape feature integration effects in visual agnosia for biological kinds PMID- 10373287 TI - Visual P300 in treatment-resistant schizophrenic patients PMID- 10373290 TI - PCNA and Ki67 proliferation markers as criteria for prediction of clinical behaviour of melanocytic tumours in cats and dogs. AB - Melanocytic tumours in domestic animals vary from benign to highly malignant. Diagnosis and prognosis are mainly based on the somewhat uncertain interpretation of cytological signs of malignancy in histological sections. The purpose of the present retrospective study was to compare the accuracy of prognosis based on the classical morphological criteria with that provided by quantitative computerized analysis of proliferation-associated antigens. Twenty-seven melanocytic tumours (20 canine and seven feline) from 24 animals (20 dogs and four cats) were examined. These comprised eight tumours histologically classified as benign melanoma (BM) (seven canine, one feline), 16 classified as primary malignant melanoma (PMM) (13 canine, three feline) and three classified as metastatic melanoma (MMM) (three feline). Tumour size, predominant histological cell type, invasive growth and clinical outcome were recorded for each case. In addition, the proliferation (phase) index and growth fraction were measured, after bleaching, by means of quantitative image analysis of tissue sections immunolabelled for proliferating cell nuclear antigen (PCNA) and MIB-1 (Ki67). Lesions classified histologically as benign or malignant, differed significantly (Copyright P<0.001) in respect of Ki67 and PCNA positivity. No such difference could be shown between PMM and MMM. High growth fraction (measured by the Ki67 positive cells) and macroscopic invasive growth were associated with decreased survival time (P=0.027 and P=0.024, respectively). Moreover, the established cytological classification also proved to be associated with the survival time (P<0.001). The results suggest that Ki67 is a potentially important prognostic factor in melanomas of the cat and dog. Other criteria, including tumour size, predominant cell type and intensity of PCNA expression, were not of significant prognostic value (P>0.05). PMID- 10373289 TI - Experimental reproduction of severe wasting disease by co-infection of pigs with porcine circovirus and porcine parvovirus. AB - Colostrum-deprived pigs were infected intranasally with a recent isolate of porcine circovirus (PCV2) and a porcine parvovirus (PPV), both from Canadian pigs with post-weaning multisystemic wasting syndrome (PMWS). Four pigs were inoculated with PCV2 alone, three with PPV alone, five with a combined PCV2/PPV inoculum, and two with a chloroform-treated combined PCV2/PPV inoculum. Pigs were killed 21-26 days after infection and tissue samples examined for gross and microscopical lesions and for the presence of viral antigens. No clinical signs, lesions or viral antigens were detected in two uninfected control pigs or in pigs inoculated with PPV alone. One pig inoculated with PCV2 alone became dull and thin. Mild to moderate histopathological lesions containing PCV2 antigen were detected in lymphoid tissues from the pigs inoculated with PCV2 alone. Pigs given the PCV2/PPV inoculum and the chloroform-treated PCV2/PPV inoculum became dull and two died. Jaundice and hepatomegaly were seen at post-mortem examination of most of the dually infected pigs. The latter showed large amounts of PCV2 antigen in numerous tissues; PPV antigen, which was less abundant, was detected in a few tissues, especially kidney. The lesions were similar to those seen in recently described field cases of porcine PMWS in North America and Europe. PMID- 10373291 TI - The role of pulmonary intravascular macrophages in the pathogenesis of African horse sickness. AB - African horse sickness (AHS) is a disease of equids, characterized by severe pulmonary oedema and caused by an orbivirus. To determine the role of pulmonary intravascular macrophages (PIMs) in the development of pulmonary microvascular changes in this disease, five horses were given an intravenous inoculation of 10(6)TCID50of serotype 4 of AHS virus. Viral replication was detected in endothelial cells, PIMs, interstitial macrophages and fibroblasts. Alveolar and interstitial oedema, and changes in pulmonary microvasculature, consisting mainly of the sequestration of neutrophils and the formation of platelet aggregates and fibrinous microthrombi, were related to endothelial changes and to a high degree of PIM activation. This suggested that the PIMs, once activated, contributed to these vascular changes by releasing chemical inflammatory mediators. PMID- 10373292 TI - Hydranencephaly and cerebellar hypoplasia in two kittens attributed to intrauterine parvovirus infection. AB - Six weeks after vaccination with modified live feline parvovirus vaccine, a cat gave birth to five kittens, three of which died soon afterwards. The remaining two kittens (A and B) survived, but at 8 weeks of age were unable to walk and showed abnormal behaviour, with lack of menace and oculovestibular responses, and severe dysmetria. These signs suggested multifocal disease associated with the cerebrum and cerebellum. Magnetic resonance imaging demonstrated severe bilateral (kitten A) or unilateral (kitten B) hydrocephalus or hydranencephaly, combined with cerebellar agenesis (kitten A) or severe hypoplasia (kitten B). Hydranencephaly was confirmed histopathologically in both kittens. Parvovirus was isolated from the kidney of one kitten. Parvoviral DNA was amplified by the polymerase chain reaction (PCR) from paraffin wax-embedded brain of both kittens. The severe malformations observed in these kittens presumably resulted from an in utero parvovirus infection, possibly due to vaccination, that occurred late in the first, or early in the second, trimester of pregnancy. PMID- 10373293 TI - Scrapie-associated prion protein in the gastrointestinal tract of sheep with natural scrapie. AB - The scrapie-associated prion protein (PrPSc), which is closely associated with scrapie infectivity, accumulates in the brain and lymphoid tissues of sheep with natural scrapie. The most probable portal of entry of the scrapie agent in sheep is the alimentary tract; little attention, however, has been paid to the gastro intestinal tract in scrapie research. In this study, we examined the presence and distribution of PrPSc within the gastro-intestinal tract of sheep with natural scrapie and scrapie-negative sheep. It was found that PrPSc accumulated in the enteric nervous system (ENS) of all scrapie-infected sheep but not in scrapie negative sheep. The distribution of PrPSc within the ENS was then studied along the entire gastro-intestinal tract in seven scrapie-infected sheep carrying various PrP genotypes. In sheep with the highest genetically determined susceptibility to scrapie, PrPSc was detected in the ENS from the oesophagus to the rectum. In sheep with a lower genetic susceptibility to scrapie, PrPSc was present in the ENS of the forestomachs, small intestine and large intestine but not in the oesophagus. In a scrapie-negative sheep with a PrP genotype associated with scrapie resistance, no PrPSc was seen in the ENS at any site along the gastro-intestinal tract. The presence of PrPSc within the ENS of scrapie-infected sheep indicates a possible role of the ENS in the pathogenesis of natural scrapie as a portal of entry to the central nervous system. PMID- 10373294 TI - Atypical cilia in the tracheal epithelium of healthy water buffaloes (Bubalus bubalis). AB - Samples of tracheal mucosa were obtained from 10 healthy adult water buffaloes and 50 000 cilia were examined ultrastructurally. Ciliary abnormalities were found in all 10 subjects. Atypical cilia occurred as compound cilia (up to 1.5%), intracytoplasmic cilia (0.07%) and swollen cilia (0.05%). The microtubular pattern was determined in 5000 cross-sectioned cilia, with about 7% showing axonemal abnormalities in which peripheral defects prevailed. Some electron-dense plugs appeared inside the cylinder lumen of 2.5% of basal bodies. Freeze-fracture studies revealed a ciliary necklace composed of up to eight rows of intramembranous particles. This fine detail appeared to differ from that of other small and large ruminants. PMID- 10373295 TI - Arteriovenous malformation of the cervical spinal cord in a dog. AB - An 8-year-old female German Shepherd dog showed first order Horner's syndrome associated with progressive right-sided hemiplegia and mega-oesophagus. Intramedullary and leptomeningeal arteriovenous malformation (AVM) was identified in the cervical spinal cord. The morphological characteristics were arteriovenous shunting, intramedullary multiple thromboses and haemorrhage, non-inflammatory necrosis of white and grey matter around the shunt, and intervening neural gliosis with neovascularization. These findings suggested that the malformation induced a focal circulatory disturbance within the cervical spinal cord and that fatal thrombosis was responsible for the sudden onset of the nervous signs and progressive neurological deterioration. This is the first report of intramedullary spinal AVM in a dog. PMID- 10373296 TI - Primary malignant histiocytosis of the brain in a dog. AB - Malignant histiocytosis is a well-recognized canine tumour, occurring primarily in Bernese mountain dogs and characterized by disseminated histiocytic infiltration of multiple visceral organs. This report describes the light microscopical and ultrastructural features of a neoplasm composed of malignant histiocytes and confined to the brain. A poorly demarcated mass in the right parieto-occipital lobe of a miniature schnauzer was composed of loosely aggregated, pleomorphic cells with abundant eosinophilic cytoplasm, expanding the meninges. Many binucleated and multinucleated giant cells and mitotic figures were seen. Immunohistochemically, the tumour cells reacted intensely for lysozyme. Ultrastructurally, the neoplastic cells had features of histiocytic cells with abundant lysosomes. The findings in this case were strikingly similar to those of disseminated malignant histiocytosis described in other dog breeds. PMID- 10373297 TI - Large granular lymphocyte pleocytosis in the cerebrospinal fluid of a dog with necrotizing meningoencephalitis. AB - The cerebrospinal fluid of a dog with necrotizing meningoencephalitis showed pleocytosis, produced mostly by large granular lymphocytes. The presence of such lymphocytes in the cerebrospinal fluid of dogs with neurological disease appears not to have been reported previously. PMID- 10373298 TI - New embryological evidence for the formation of quadricuspid aortic valves in the Syrian hamster (Mesocricetus auratus). AB - A Syrian hamster embryo, aged 11 days and 4 h post-coitus, had a developing quadricuspid aortic valve. The septation of the cardiac outflow tract was confined to the distal part of the conotruncus. There was a conspicuous recess in the anlage that normally gives rise to the left aortic valve cushion. Globular endothelial cells arranged in several layers were present at the luminal side of the recess. The present findings support the hypothesis that in the Syrian hamster, quadricuspid aortic valves result from the division of one of the three mesenchymal anlagen that give rise to normal aortic valves. In addition, they indicate that the division of the anlage is due to the invagination of the endothelial layer that covers its luminal side. The invagination of the endothelium starts at a very early stage of the valvulogenesis, namely, when the conotruncal ridges begin to fuse at the distal portion of the embryonic cardiac outflow tract. PMID- 10373299 TI - Simultaneous seminoma and interstitial cell tumour in a rabbit with a previous cutaneous basal cell tumour. AB - The development of spontaneous multiple tumours is a rare event in domestic rabbits. The diagnosis of a cutaneous basal cell tumour and the successive development of simultaneous bilateral testicular tumours with dissimilar histology (a seminoma and an interstitial cell tumour) are described in a vasectomized, crossbred dwarf rabbit, aged 6 years. Two cases of basalioma associated with uterine adenocarcinoma have been previously described in rabbits. A similar association between basal cell neoplasia and development of tumours (e.g., testicular and breast cancer) at cutaneous and non-cutaneous sites has been reported in man. PMID- 10373300 TI - The cortical granule serine protease CGSP1 of the sea urchin, Strongylocentrotus purpuratus, is autocatalytic and contains a low-density lipoprotein receptor-like domain. AB - Trypsin-like activity is secreted from eggs of many species at fertilization, and this activity is believed to be critical for the block to polyspermy. Here we show that a cortical granule serine protease of sea urchins is the major and perhaps only protease family member important for fertilization. Zymography assays suggest that the cortical granules contain a single serine protease that can undergo autocatalysis and is secreted upon egg activation. We used this finding to identify a cDNA clone from a Strongylocentrotus purpuratus ovary cDNA library that encodes a 581-amino-acid-residue protein that we refer to as cortical granule serine protease 1 (CGSP1). The catalytic domain of the protein contains the essential residues of the catalytic triad characteristic of a member of the trypsin-like family of serine proteases and the N-terminus of CGSP1 resembles the ligand-binding domain of the low-density lipoprotein (LDL) receptor. Antibodies raised separately to both the protease and LDL receptor-like domains each localize to the cortical granules of unfertilized eggs. Furthermore, the full-length form of CGSP1, as well as intermediate and active forms of the protease, is detected in cortical granules by immunoblot analysis. Our evidence suggests that CGSP1 is activated at fertilization and is responsible for the protease-mediated reactions that follow cortical granule exocytosis and contribute to the block to polyspermy. PMID- 10373301 TI - Impact of node ablation on the morphogenesis of the body axis and the lateral asymmetry of the mouse embryo during early organogenesis. AB - The node of the mouse gastrula is the major source of the progenitor cells of the notochord, the floor plate, and the gut endoderm. The node may also play a morphogenetic role since it can induce a partial body axis following heterotopic transplantation. The impact of losing these progenitor cells and the morphogenetic activity on the development of the body axes was studied by the ablation of the node at late gastrulation. In the ablated embryo, an apparently intact anterior-posterior body axis with morphologically normal head folds, neural tube, and primitive streak developed during early organogenesis. Cell fate analysis revealed that the loss of the node elicits de novo recruitment of neural ectoderm and somitic mesoderm from the surrounding germ-layer tissues. This leads to the restoration of the neural tube and the paraxial mesoderm. However, the body axis of the embryo was foreshortened and somite formation was retarded. Histological and gene expression studies reveal that in most of the node-ablated embryos, the notochord in the trunk was either absent or interrupted, and the floor plate was absent in the ventral region of the reconstituted neural tube. The loss of the node did not affect the differentiation of the gut endoderm or the formation of the mid- and hindgut. In the node-ablated embryo, expression of the Pitx2 gene in the lateral plate mesoderm was no longer restricted to the left side but was found on both sides of the body or was completely absent from the lateral plate mesoderm. Therefore, the loss of the node results in the failure to delineate the laterality of the body axis. The node and its derivatives therefore play a critical role in the patterning of the ventral neural tube and lateral body axis but not of the anterior-posterior axis during early organogenesis. PMID- 10373302 TI - The process of pharynx regeneration in planarians. AB - To understand the cellular events during planarian regeneration, we analyzed the process of pharynx regeneration in both head and tail pieces using cell-type specific markers. Interestingly, cells expressing the pharynx-muscle-specific myosin heavy chain gene (DjMHC-A) appeared within 24 h after amputation (prior to the formation of a pharynx rudiment) in the mesenchymal space of the stump, not in the blastema region. These DjMHC-A-positive cells migrated to the midline and formed the pharynx rudiment. Even after formation of the pharynx rudiment, DjMHC A-positive cells constantly appeared in the mesenchymal space in the region surrounding the pharynx rudiment and participated in the growth of the pharynx rudiment. These observations clearly indicated that the cells involved in pharynx muscle formation are committed in the mesenchymal space of the stump, rather than in the blastema region or the pharynx rudiment during planarian regeneration. We also analyzed the process of regeneration of the pharynx epithelia using a monoclonal antibody and investigated the origin of the pharynx epithelia. PMID- 10373303 TI - The even-skipped locus is contained in a 16-kb chromatin domain. AB - The even-skipped (eve) gene of Drosophila melanogaster is a crucial member of the pair-rule class of segmentation genes. We report here the characterization of a 16-kb region sufficient for all known aspects of eve expression and the rescue of an eve null mutation. We began by examining 45 kb surrounding the eve coding sequence for DNaseI hypersensitive sites and other transcription units. We find that the previously identified eve regulatory elements, those for early stripes 2, 3, and 7 and the late element, do not generate prominent hypersensitive sites. However, strong, constitutive DNaseI hypersensitive sites flank a 16-kb region, within which one developmentally regulated site is found at the eve promoter region. P-element transformation of this 16-kb domain into eve mutants rescues them to adult viability. This 16-kb domain contains regulatory elements for all known features of eve expression: the seven major blastoderm stripes, minor stripe expression during germ band extension, and later expression in the lateral mesodermal muscle precursor cells, in the central nervous system, adjacent to the invaginating proctodeum, and in a ring around the anal pad. We have begun a preliminary dissection of the 16-kb domain into its constituent regulatory elements. Other major findings include the following: (1) There is a second element for late stripe expression adjacent to the traditional late element. (2) A stripe element 3' of the gene interacts with the late element to give rise to the minor stripes seen in the even-numbered parasegments. (3) Expression in the proctodeum and anal pad is driven by sequences both 5' and 3' of the gene. (4) Expression in different sites in the central nervous system is driven by separable elements widely dispersed throughout 8 kb 3' of the gene. PMID- 10373304 TI - Dorsal-ventral patterning defects in the eye of BF-1-deficient mice associated with a restricted loss of shh expression. AB - Brain factor 1 (BF-1) is a winged-helix transcription factor with restricted expression in the anterior optic vesicle and in the telencephalic neuroepithelium of the neural tube. We have previously found that targeted disruption of the BF-1 gene results in hypoplasia of the cerebral hemispheres, which is more severe in structures derived from the ventral telencephalon. Here we show that the loss of BF-1 leads to multiple developmental anomalies of the eyes. The most ventral structure arising from the optic vesicle, the optic stalk, is missing and is replaced by an expanded retina. Ventral closure of the optic cup and choroid fissure does not occur. These dorsal-ventral patterning defects are not limited to the BF-1-expressing (anterior) cells, but also involve the cells of the posterior optic vesicle. Sonic hedgehog (shh) expression within the ventral telencephalic neuroepithelium is specifically lost in the BF-1(-/-) mutant. Taken together, these findings suggest that shh produced at this site plays a role in patterning the developing eye. This localized deficit in shh expression may also contribute to the prominence of the ventral defects in the telencephalon of the BF-1(-/-) mutant. PMID- 10373306 TI - Apextrin, a novel extracellular protein associated with larval ectoderm evolution in Heliocidaris erythrogramma. AB - During the evolution of direct development in the sea urchin Heliocidaris erythrogramma major modifications occurred, which allowed the precocious formation of adult-specific structures and led to a novel larval body that surrounds these structures. The HeET-1 gene was isolated in a differential screen for transcripts enriched in the early embryos of H. erythrogramma relative to those of its indirect-developing congener, H. tuberculata. HeET-1 was unique among the three genes found in that no homologous transcript was detected in H. tuberculata total embryonic RNA blots. To verify this apparently extreme differential expression of the HeET-1 genes in Heliocidaris, we isolated the HeET 1 homologue from H. tuberculata genomic DNA and used it to probe blots of poly(A)+ RNA prepared from H. tuberculata embryos. It is expressed in H. tuberculata embryos at levels undetectable by this technique. The predicted amino acid sequence of HeET-1 suggested that it encodes a novel secreted protein. To assess the function of HeET-1, we raised polyclonal antisera to the HeET-1 encoded protein. We find that it is present in eggs in a type of secretory vesicle and that this maternal pool is gradually secreted after fertilization. As cells acquire apical-basal polarity in the blastula the protein becomes localized to the apical extracellular matrix, leading us to name the protein apextrin. The apical extracellular localization of apextrin is maintained in the columnar cells of the larval ectoderm until their internalization at metamorphosis. Ingressing mesenchyme cells rapidly endocytose apextrin upon leaving the vegetal plate. Comparison with fibropellin III, an apical lamina component, suggests that apextrin is an extracellular protein that is in tighter association with the plasma membrane than is the hyalin layer or apical lamina. We propose that apextrin is involved in apical cell adhesion and that its high level of expression may represent an adaptive cooption necessary for strengthening the large H. erythrogramma embryo. PMID- 10373305 TI - Novel strategy yields candidate Gsh-1 homeobox gene targets using hypothalamus progenitor cell lines. AB - We describe the successful application of a strategy that potentially provides for an efficient and universal screen for downstream gene targets. We used the promoter of the Gsh-1 homeobox gene to drive expression of the SV40 T-antigen gene in transgenic mice. We have previously shown that the Gsh-1 homeobox gene is expressed in discrete domains of the ganglionic eminences, diencephalon, and hindbrain during brain development. Gsh-1-SV40 T transgenic mice showed cellular hyperplasia in regions of the brain coincident with Gsh-1 expression. The Gsh-1 SV40 T transgene was introduced, by breeding, into Gsh-1 homozygous mutant mice, and Gsh-1 -/- cell lines were made. Clonal cell lines were generated and analyzed by Northern blot hybridizations and Affymetrix GeneChip probe arrays to determine gene expression profiles. The results indicate that the cell lines remain representative of early developmental stages. Further, immunocytochemistry showed uniformly high levels of nestin expression, typical of central nervous system progenitor cells, and the absence of terminal differentiation markers of neuronal cells. One clonal cell line, No. 14, was then stably transfected with a tet inducible Gsh-1 expression construct and subcloned. The starting clone 14, together with the uninduced and induced subclones, provided cell populations with varying levels of Gsh-1 expression. Differential display and Affymetrix GeneChip probe arrays were then used to identify transcript differences that represent candidate Gsh-1 target genes. Of particular interest, the drm and gas1 genes, which repress cell proliferation, were observed to be activated in Gsh-1 expressing cells. These observations support models predicting that homeobox genes function in the regulation of cell proliferation. PMID- 10373307 TI - Socket cells mediate spicule morphogenesis in Caenorhabditis elegans males. AB - Caenorhabditis elegans male spicule morphogenesis requires the coordinated cellular behaviors of several types of cells. We found that the spicule neurons and sheath cells, although important for spicule function, are dispensable for spicule morphology. In contrast, the spicule socket cells are essential for both spicule elongation and formation of spicule cuticle. The socket cells are not only necessary but also sufficient to produce spicule cuticle. This functional aspect of socket cells is genetically separable from their function in mediating spicule elongation: elongated spicules with defective spicule cuticle can be formed. During spicule morphogenesis, the expression of an egl-17::GFP reporter gene is found in the spicule socket cells and its expression appears to be regulated in the socket cells. Mutants defective in TGF-beta signaling display a crumpled spicules phenotype as a result of failure of socket cell movement during spicule morphogenesis. These observations suggest that both the FGF and the TGF beta signaling pathways might be involved in spicule elongation. PMID- 10373308 TI - Chamber-specific cardiac expression of Tbx5 and heart defects in Holt-Oram syndrome. AB - To further define the role of a T-box transcription factor, Tbx5, in cardiac development, we have examined its expression in the developing mouse and chick heart and correlated this pattern with cardiac defects caused by human TBX5 mutations in Holt-Oram syndrome. Early in the developing heart, Tbx5 is uniformly expressed throughout the entire cardiac crescent. Upon formation of the linear heart tube, Tbx5 is expressed in a graded fashion, stronger near the posterior end and weaker at the anterior end. As the heart tube loops, asymmetric Tbx5 expression continues; Tbx5 is expressed in the presumptive left ventricle, but not the right ventricle or outflow tract. This pattern of expression is maintained in more mature hearts. Expression in the ventricular septum is restricted to the left side and is contiguous with left ventricular free wall expression. Trabeculae, vena cavae (inferior and superior), and the atrial aspect of the atrioventricular valves also express high levels of Tbx5. These patterns of Tbx5 expression provide an embryologic basis for the prevalence of atrial septal defects (ostium primum and secundum), ventricular muscular septal defects, and left-sided malformations (endocardial cushion defects, hypoplastic left heart, and aberrant trabeculation) observed in patients with Holt-Oram syndrome. PMID- 10373309 TI - Spag4, a novel sperm protein, binds outer dense-fiber protein Odf1 and localizes to microtubules of manchette and axoneme. AB - Outer dense fibers are structures unique to the sperm tail. No definite function for these fibers has been found, but they may play a role in motility and provide elastic recoil. Their composition had been described before, but only two of the fiber proteins, Odf1 and Odf2, are cloned. We cloned Odf2 by virtue of its functional and specific interaction with Odf1, which, we show, is mediated by a leucine zipper. Further work demonstrated that the 84-kDa Odf2 protein localizes to both the cortex and the medulla of the fibers, whereas the 27-kDa Odf1 protein is present only in the medulla. Here we report the cloning and characterization of a new Odf1-interacting protein, Spag4. Spag4 mRNA is spermatid specific, and the 49-kDa Spag4 protein complexes specifically with Odf1, but not Odf2, mediated by a leucine zipper. It also self-associates. In contrast to Odf1 and Odf2, Spag4 protein localizes to two microtubule-containing spermatid structures. Spag4 is detectable in the transient manchette and it is associated with the axoneme in elongating spermatids and epididymal sperm. Our data suggest a role for Spag4 in protein localization to two major sperm tail structures. PMID- 10373310 TI - Embryonic lens repels retinal ganglion cell axons. AB - During development of the vertebrate visual system, retinal ganglion cell (RGC) axons follow a precise path toward their midbrain targets. Although much is known about the cues that direct RGC axons once they have left the optic disc, less is known about the guidance of axons at earlier stages, when RGCs first send out their axons to navigate within the developing retina. Using collagen gel coculture experiments, we find that the embryonic lens produces a powerful diffusible repulsive activity for RGC axons. We also find that this activity is localized to the lens epithelium and not the lens fiber layer, while the pigmented epithelium and vitreous humour are devoid of activity. The further observation that the lens also chemorepels primary sensory axons, but does not repel olfactory bulb axons, shows that this activity is specific for subsets of axons. Our experiments have excluded two candidate repellents for RGC axons (collapsin-1/sema III and chondroitin sulfate proteoglycans). These results implicate the lens in the earliest stages of RGC axon guidance. One function of the lens repellent may be to prevent aberrant targeting toward the lens, and it may also be involved in the directional guidance of RGC axons toward the optic disc. PMID- 10373311 TI - FGF7 and FGF10 directly induce the apical ectodermal ridge in chick embryos. AB - During vertebrate limb development, the apical ectodermal ridge (AER) plays a vital role in both limb initiation and distal outgrowth of the limb bud. In the early chick embryo the prelimb bud mesoderm induces the AER in the overlying ectoderm. However, the direct inducer of the AER remains unknown. Here we report that FGF7 and FGF10, members of the fibroblast growth factor family, are the best candidates for the direct inducer of the AER. FGF7 induces an ectopic AER in the flank ectoderm of the chick embryo in a different manner from FGF1, -2, and -4 and activates the expression of Fgf8, an AER marker gene, in a cultured flank ectoderm without the mesoderm. Remarkably, FGF7 and FGF10 applied in the back induced an ectopic AER in the dorsal median ectoderm. Our results suggest that FGF7 and FGF10 directly induce the AER in the ectoderm both of the flank and of the dorsal midline and that these two regions have the competence for AER induction. Formation of the AER of the dorsal median ectoderm in the chick embryo is likely to appear as a vestige of the dorsal fin of the ancestors. PMID- 10373312 TI - Identification of Rab3A GTPase as an acrosome-associated small GTP-binding protein in rat sperm. AB - The acrosome reaction is a membrane fusion event that is prerequisite for sperm penetration through the zona pellucida. To elucidate the molecular mechanisms involved in membrane fusion, the expression and localization of Rab proteins, a subfamily of small GTPases that have been shown to play key roles in regulation of intracellular membrane traffic and exocytosis, were examined in rat testis and sperm. Reverse transcription polymerase chain reaction, immunoblot analysis, and immunofluorescence microscopy revealed that Rab3A protein, which is thought to be involved in regulation of exocytosis in neurons and endocrine cells, is associated with the sperm acrosome. The protein was undetectable in acrosome-free heads prepared by sucrose density gradient centrifugation. Immunogold electron microscopy performed on ultrathin cryosections provided further evidence that Rab3A protein is associated with the acrosomal membrane. Acrosome reaction assays revealed that synthetic peptide of the Rab3 effector domain inhibited acrosomal exocytosis triggered by calcium ionophore A23187 in a concentration-dependent fashion, suggesting that Rab3A acts as an inhibitory regulator in the acrosome reaction. In view of the putative role of Rab3A protein in membrane fusion systems, these results suggest that Rab3A could be involved in regulating the mammalian acrosome reaction by controlling the membrane fusion system in sperm. PMID- 10373313 TI - Editorial. PMID- 10373314 TI - Special Issue on the Math-Fact Retrieval Hypothesis. PMID- 10373316 TI - Sex Differences in Mathematical Abilities: Commentary on the Math-Fact Retrieval Hypothesis. AB - The hypothesis that a male advantage in speed of math-fact retrieval underlies the sex difference, favoring males, in mathematical abilities is unique and provocative. However, the hypothesis does not provide an explanation for the male advantage in mathematical domains, such as geometry, that do not require arithmetic, nor does it accommodate the sex differences in social and occupational interests that contribute to the sex difference in mathematical achievement. An alternative hypothesis focusing on the sex difference in three dimensional spatial cognition is favored over the math-fact retrieval hypothesis. Copyright 1999 Academic Press. PMID- 10373315 TI - Math-Fact Retrieval as the Cognitive Mechanism Underlying Gender Differences in Math Test Performance. AB - Males from select populations receive better scores on standardized math achievement tests than females. The research reported in this article evaluates the hypothesis that the reason for these differences is that males are faster at retrieving basic math facts. Studies 1-3 demonstrate that math-fact retrieval predicts performance on math achievement tests with students in grades 5-8 and in college. Studies 4-6 show that males and females in grades 2-8 and in college have different patterns of math-fact retrieval performance and that males at the high positive end of the retrieval distribution are faster than comparable females. Study 5 also demonstrates that math-fact retrieval varies in three populations (Anglo-American, Chinese-American, Hong Kong Chinese) and that speed of retrieval improves with practice. Studies 7-9 tested the hypothesis that males are faster than females on retrieval tasks in general. Study 7 showed that there were no gender differences on simple retrieval tasks, and Studies 8 and 9 showed that females were slightly faster than males on verbal-processing tasks. The General Discussion indicates that the math-fact retrieval hypothesis is consistent with previous research. It also relates the math-fact retrieval hypothesis to theories of cognitive performance and introduces the practice and engagement hypothesis. This hypothesis explains the origin of gender differences in math and reading and relates those differences to the existing literature on gender differences in academic performance. The article concludes with a description of needed future research and a discussion of the educational implications of the math-fact retrieval hypothesis. Copyright 1999 Academic Press. PMID- 10373317 TI - Does Math-Fact Retrieval Explain Sex Differences in Mathematical Test Performance? A Commentary. AB - We focus on four issues in commenting on Royer et al.'s work testing their hypothesis that sex differences in test performance can be explained by differences in math-fact retrieval latency. The first issue is whether the studies conducted by Royer et al. directly test their hypothesis; we argue that they do not. The second issue involves questions about the strength of the gender differences in math-fact retrieval. The third is whether math-fact retrieval should be considered the major mechanism in explaining the differences in test performance. In considering this third issue we discuss other theories regarding the nature of math problem solving that are useful for understanding test performance differences between males and females. The fourth issue is what particular experiential differences boys and girls have in math classrooms could contribute to the sex differences in test performance. We also consider sex differences in math attitudes and motivation. Copyright 1999 Academic Press. PMID- 10373318 TI - Reply to the Commentaries on the Math-Fact Retrieval Hypothesis. AB - Geary and Wigfield and Brynes (this issue) point out a number of limitations of the math-fact retrieval hypothesis that we agree with. For instance, we acknowledge that whereas the correlational evidence we offer in our article (this issue) provides suggestive evidence for a link between math-fact retrieval and gender differences in math test performance, that evidence is not compelling. We also acknowledge that even if it is the case that math-fact retrieval is one of the cognitive mechanisms responsible for the gender differences in math performance, there are still many aspects of gender differences in math performance that need to be understood. We also point out a number of areas where we disagree. Most prominently, we do not believe that the spatial cognition hypothesis or affective/motivational hypotheses account for two significant literatures-gender differences in test performance and gender differences in grade performance. We discuss the basis for our beliefs and close with a discussion of the need for intervention research that will resolve some of the issues discussed in the series of articles in this issue. At the end of the article we also present a very speculative hypothesis that would knit together all of the positions presented in the articles in this issue of CEP. Copyright 1999 Academic Press. PMID- 10373319 TI - Zebrafish genetic map with 2000 microsatellite markers. AB - The zebrafish is the first vertebrate organism used for large-scale genetic screens seeking genes critical to development. These screens have been quite successful, with more than 1800 recessive mutations discovered that speak to morphogenesis of the vertebrate embryo. The cloning of the mutant genes depends on a dense genetic map. The 2000 markers we present here, using microsatellite (CA) repeats, provides 1.2-cM average resolution. One centimorgan in zebrafish is about 0. 74 megabase, so, for many mutations, these markers are close enough to begin positional cloning by YAC walks. PMID- 10373320 TI - Genetic dissection of "OLETF," a rat model for non-insulin-dependent diabetes mellitus: quantitative trait locus analysis of (OLETF x BN) x OLETF. AB - To identify genetic determinants relevant to non-insulin-dependent diabetes mellitus (NIDDM), we performed a genome-wide analysis for quantitative trait loci (QTLs) using 359 backcross progeny of the Otsuka Long-Evans Tokushima Fatty (OLETF) rat. The OLETF strain is a well-studied animal model of obese NIDDM, with features of hyperinsulinemia, hyperglycemia, insulin resistance, and abundant abdominal fat. Our extensive genomic scanning with 218 markers revealed nine significant QTLs, including a strong determinant of obesity on chromosome 1 (Dmo1: LOD = 13.99, for body weight). Two highly significant QTLs for glucose homeostasis were found, one on chromosome 1 (Dmo4 LOD = 7.16, for postprandial glucose level) and the other on chromosome X (Dmo11/Odb1: LOD = 7.81, for postprandial glucose level). These data are comparable to results of our previous studies of the OLETF rat. PMID- 10373321 TI - Isolation and mapping of novel candidate genes for retinal disorders using suppression subtractive hybridization. AB - We have constructed human cDNA libraries enriched for retina- and retinal pigment epithelium (RPE)/choroid-specific cDNAs through suppression subtractive hybridization. The sequence of 314 cDNAs from the retina enriched library and 126 cDNAs from the RPE/choroid enriched library was analyzed. Based on the absence of a database match, 25% of the retina cDNA clones and 16% of the RPE/choroid cDNA clones are novel cDNAs. The expression profiles of 86 retina and 21 RPE/choroid cDNAs were determined by a semiquantitative reverse transcription polymerase chain reaction technique. Thirty-three cDNAs were expressed exclusively or most prominently in retina or RPE/choroid. These cDNAs were mapped in the human genome by radiation hybrid mapping. Eleven cDNAs colocalized with loci involved in retinal disorders. One cDNA mapped in a 1.5-megabase critical region for autosomal recessive retinitis pigmentosa (RP12). Another cDNA was assigned to the 7.7-cM RP17 linkage interval. Seven cDNAs colocalized with four loci involved in Bardet-Biedl syndrome. PMID- 10373322 TI - A modular, positive selection bacterial artificial chromosome vector with multiple cloning sites. AB - To construct large-insert libraries for the sequencing, mapping, and functional studies of complex genomes, we have constructed a new modular bacterial artificial chromosome (BAC) vector, pBACe3.6 (GenBank Accession No. U80929). This vector contains multiple cloning sites located within the sacB gene, allowing positive selection for recombinant clones on sucrose-containing medium. A recognition site for the PI-SceI nuclease has also been included, which permits linearization of recombinant DNA irrespective of the characteristics of the insert sequences. An attTn7 sequence present in pBACe3.6 permits retrofitting of BAC clones by Tn7-mediated insertion of desirable sequence elements into the vector portion. The ability to retrofit BAC clones will be useful for functional analysis of genes carried on the cloned inserts. The pBACe3.6 vector has been used for the construction of many genomic libraries currently serving as resources for large-scale mapping and sequencing. PMID- 10373323 TI - A new tool for the rapid cloning of amplified and hypermethylated human DNA sequences from restriction landmark genome scanning gels. AB - Restriction landmark genome scanning (RLGS) is an effective genome-scanning technique capable of identifying DNA amplification and aberrant DNA methylation. Previously published methods for the cloning of human DNA fragments from RLGS gels have been successful only for high-copy-number fragments (repetitive elements or DNA amplifications). We present here the first technique capable of efficiently cloning single-copy human DNA fragments ("spots") identified in RLGS profiles. This technique takes advantage of a plasmid-based, human genomic DNA, NotI/EcoRV boundary library. The library is arrayed in microtiter plates. When clones from a single plate are pooled and mixed with genomic DNA, the resultant RLGS gel is a normal profile with a defined set of spots showing enhanced intensity for that particular plate. This was performed for a set of 32 plates as well as their pooled rows and columns. Thus, we have mapped individual RLGS spots to exact plate, row, and column addresses in the library and have thereby obtained immediate access to these clones. The feasibility of the technique is demonstrated in examples of cloning methylated DNA fragments identified in human breast tumor and testicular tumor RLGS profiles and in the cloning of an amplified DNA fragment identified in a human medulloblastoma RLGS profile. PMID- 10373324 TI - Gene structure, chromosomal location, and expression pattern of maleylacetoacetate isomerase. AB - The gene for maleylacetoacetate isomerase (MAAI) (EC 5.2.1.2) was the last gene in the mammalian phenylalanine/tyrosine catabolic pathway to be cloned. We have isolated the human and murine genes and determined their genomic structure. The human gene spans a genomic region of approximately 10 kb, has 9 exons ranging from 50 to 528 bp in size, and was mapped to 14q24.3-14q31.1 using fluorescence in situ hybridization. The complete catabolic pathway of phenylalanine/tyrosine is normally restricted to liver and kidney, but the maleylacetoacetate isomerase gene is expressed ubiquitously. This suggests a possible second role for the MAAI protein different from phenylalanine/tyrosine catabolism. We have searched for mutations in the maleylacetoacetate isomerase gene in four cases of unexplained severe liver failure in infancy with clinical similarities to hereditary tyrosinemia type I (pseudotyrosinemia). Several amino acid changes were identified, but all were found to retain MAAI activity and thus represent protein polymorphisms. We conclude that MAAI deficiency is not a common cause of the pseudotyrosinemic phenotype. PMID- 10373325 TI - Identification and chromosomal location of two human genes encoding enzymes potentially involved in proteolytic maturation of farnesylated proteins. AB - Two human cDNAs encoding proteins similar to yeast enzymes involved in proteolytic processing of farnesylated proteins like a-factor mating pheromone and Ras2p have been cloned from an ovary cDNA library. These proteins have been tentatively called Face-1 and Face-2 (farnesylated protein-converting enzymes 1 and 2), respectively, and are integral membrane proteins, belonging to distinct families of metalloproteinases. Northern blot analysis of poly(A)+ RNAs isolated from a wide variety of human tissues demonstrated that both genes are expressed in all examined tissues, which suggests that these enzymes play housekeeping roles in normal processes. Fluorescence in situ hybridization experiments showed that the human FACE-1 gene maps to 1p34, whereas FACE-2 is located at 11q13, a region frequently amplified in human carcinomas and lymphomas. On the basis of these results, we suggest that inhibition of Face-1 and/or Face-2 could be part of strategies directed to block the functioning of prenylated proteins activated in oncogenic processes, including Ras proteins. PMID- 10373326 TI - Characterization of novel promoter and enhancer elements of the mouse homologue of the Dent disease gene, CLCN5, implicated in X-linked hereditary nephrolithiasis. AB - The murine homologue of the human chloride channel gene, CLCN5, defects in which are responsible for Dent disease, has been cloned and characterized. We isolated the entire coding region of mouse Clcn5 cDNA and approximately 45 kb of genomic sequence embracing the gene. To study its transcriptional control, the 5' upstream sequences of the mouse Clcn5 gene were cloned into a luciferase reporter vector. Deletion analysis of 1.5 kb of the 5' flanking sequence defined an active promoter region within 128 bp of the putative transcription start site, which is associated with a TATA motif but lacks a CAAT consensus. Within this sequence, there is a motif with homology to a purine-rich sequence responsible for the kidney-specific promoter activity of the rat CLC-K1 gene, another member of the chloride-channel gene family expressed in kidney. An enhancer element that confers a 10- to 20-fold increase in the promoter activity of the mouse Clcn5 gene was found within the first intron. The organization of the human CLCN5 and mouse Clcn5 gene structures is highly conserved, and the sequence of the murine protein is 98% similar to that of human, with its highest expression seen in the kidney. This study thus provides the first identification of the transcriptional control region of, and the basis for an understanding of the regulatory mechanism that controls, this kidney-specific, chloride-channel gene. PMID- 10373327 TI - Orthologues of the Caenorhabditis elegans longevity gene clk-1 in mouse and human. AB - The clk-1 gene was isolated from the long-lived mutant of Caenorhabditis elegans and was suggested to play a biological role in longevity (Ewbank et al., 1997, Science 275: 980-983). The primary structure of CLK-1 showed a significant homology to Saccharomyces cerevisiae Coq7p/Cat5p, which is required for the biosynthesis of ubiquinone and the derepression of gluconeogenic genes. In the present study, we isolated and characterized human and mouse orthologues of the COQ7/CLK-1 gene. Sequence analysis of both the human and the mouse COQ7 cDNAs showed an open reading frame composed of 217 amino acids with calculated molecular mass of 24,309 and 24,044 Da, respectively. Homology search revealed that human COQ7 showed 85% identity to mouse COQ7, 89% identity to rat COQ7, 53% identity to C. elegans CLK-1, and 37% identity to S. cerevisiae Coq7p/Cat5p. Zoo blot analysis implied that the COQ7 gene was well conserved among mammal, bird, and reptile genomes. Tissue blot analysis showed that human COQ7 is dominantly transcribed in heart and skeletal muscle. Genomic analyses revealed that the human COQ7 gene is composed of six exons spanning 11 kb of human genome as a single-copy gene. Radiation hybrid mapping assigned the COQ7 gene to human chromosome 16p12.3-p13.11. PMID- 10373328 TI - Cloning, mapping, and expression of two novel actin genes, actin-like-7A (ACTL7A) and actin-like-7B (ACTL7B), from the familial dysautonomia candidate region on 9q31. AB - Two novel human actin-like genes, ACTL7A and ACTL7B, were identified by cDNA selection and direct genomic sequencing from the familial dysautonomia candidate region on 9q31. ACTL7A encodes a 435-amino-acid protein (predicted molecular mass 48.6 kDa) and ACTL7B encodes a 415-amino-acid protein (predicted molecular mass 45. 2 kDa) that show greater than 65% amino acid identity to each other. Genomic analysis revealed ACTL7A and ACTL7B to be intronless genes contained on a common 8-kb HindIII fragment in a "head-to-head" orientation. The murine homologues were cloned and mapped by linkage analysis to mouse chromosome 4 in a region of gene order conserved with human chromosome 9q31. No recombinants were observed between the two genes, indicating a close physical proximity in mouse. ACTL7A is expressed in a wide variety of adult tissues, while the ACTL7B message was detected only in the testis and, to a lesser extent, in the prostate. No coding sequence mutations, genomic rearrangements, or differences in expression were detected for either gene in familial dysautonomia patients. PMID- 10373329 TI - Ethnic variation in the thymidylate synthase enhancer region polymorphism among Caucasian and Asian populations. AB - Thymidylate synthase (TS) regulates the production of DNA synthesis precursors and is an important target of cancer chemotherapy. A tandem repeat sequence in a TS promoter enhancer region (TSER) was recently identified. Polymorphic variation affected in vitro expression levels of the gene. We evaluated the influence of ethnicity on TSER genotype. Allele frequency was similar in Caucasian and Southwest Asian subjects. However, homozygous triple repeat subjects were twice as common in Chinese subjects (67%) than in Caucasian subjects (38%). This demonstrates significant ethnic variation in a TS gene regulatory element which may have significant impact on pyrimidine homeostasis and drug therapy. PMID- 10373330 TI - Molecular cloning and characterization of a novel human CC chemokine, SCYA26. AB - By searching the Expressed Sequence Tag database, a full-length cDNA for a novel human CC chemokine was cloned. This cDNA encoded a 94-amino-acid protein with a putative signal peptide of 26 amino acids. The deduced mature protein had the four conserved cysteine residues characteristic of CC chemokines and showed 44% identity with MIP-1beta and 40% identity with MIP-1alpha, RANTES, and MCP-4. mRNA for this chemokine was expressed constitutively in human heart and liver and with lesser but detectable levels in skeletal muscle, kidney, and small intestine. To investigate its biological activity, the protein was expressed in mammalian cells and purified by affinity chromatography. The recombinant protein demonstrated chemotactic activity in vitro for T cells and monocytes but not for neutrophils. The gene was mapped to chromosome 7q11.2 by fluorescence in situ hybridization. Based on its structural identity with other CC chemokines and the chemotactic activity and chromosomal location of this chemokine, we designate this chemokine small inducible cytokine subfamily A, member 26 (SCYA26). This gene symbol has been approved by the HUGO Gene Nomenclature Committee. PMID- 10373331 TI - An integrated somatic cell hybrid, YAC, and BAC map of the Rmc1 region of mouse chromosome 1. AB - Rmc1, the cellular receptor for the polytropic class of murine retroviruses, determines the tissue tropism of the virus and therefore plays a critical role in the pathogenesis of polytropic virus-induced leukemia. Previously we reported the physical mapping of this gene to a 5-cM region of mouse chromosome 1 and the construction of a yeast artificial chromosome (YAC) contig across this region. In this report we describe the refinement of the Rmc1 candidate region to approximately 600 kb and the generation of an integrated somatic cell hybrid, YAC, and bacterial artificial chromosome contig spanning the region. A number of genes and loci were physically ordered along the chromosome, including a recently identified candidate for Rmc1. PMID- 10373332 TI - Panel description. Regional mapping panels for chromosomes 6, 9, and 16. PMID- 10373333 TI - Ovarian hormones influence territorial aggression in free-living female mountain spiny lizards. AB - Females are aggressive in many species but relatively little is known about the hormonal basis of female aggression, especially in free-living animals. Female mountain spiny lizards aggressively defend territories from other females. Previously, we showed that plasma levels of testosterone (T) and estradiol (E) are positively associated with levels of female aggression. Here, we manipulated hormone levels in free-living females and examined aggression expressed by females returned to their natural territories. Females received one of the following: (1) ovariectomy + empty implant (OVEX), (2) ovariectomy + T implant (T IMP), or (3) sham surgery + empty implant (SHAM). OVEX females had reduced plasma levels of E but not T relative to SHAM females. T-IMP females had elevated plasma levels of T. Levels of display and aggression in OVEX females were reduced relative to SHAM females. T-IMP females had restored levels of display behavior although, unlike SHAM, no T-IMP females expressed the overt aggressive behavior of charging. These data are most consistent with the hypothesis that an ovarian factor such as E promotes female aggression, since ovariectomy reduced both plasma E and aggression but had no effect on plasma T. The results from the T-IMP females are also consistent with this hypothesis if we assume that the effects of T are due to aromatization to E in target tissues. The data do not rule out a role for T in promoting female aggression since T-implants resulted in elevated plasma T and restored display behaviors. This study represents one of the first studies examining the hormonal basis of female aggression in free-living females. PMID- 10373334 TI - Reduced efficacy of 8-OH-DPAT's inhibition of lordosis behavior by prior estrogen treatment. AB - The effects of bilateral VMN infusion with the 5-HT1A receptor agonist, (+/-)-8 hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT; 200, 1000, or 2000 ng), on lordosis behavior were examined in hormonally primed ovariectomized rats. When rats were given a single injection with 25 microg estradiol benzoate followed 48 h later with 500 microg progesterone, inhibition of lordosis behavior was evident at all doses of 8-OH-DPAT. However, when rats were treated with 25 microg estradiol benzoate followed 7 days later with a second injection of 25 microg estradiol benzoate and then progesterone, none of the doses of 8-OH-DPAT effectively inhibited lordosis behavior. In some rats, cannulae were located near the most rostral portion of the VMN. In these rats, there was no effect of the second estrogen treatment on the response to 8-OH-DPAT. Therefore, a second experiment was performed to specifically evaluate the effects of two estradiol benzoate treatments on the response to bilateral 8-OH-DPAT infusion in the rostral VMN. In contrast to the reduced effectiveness of the 8-OH-DPAT infusion in the mid to caudal VMN in rats given two injections with estradiol benzoate, 2000 ng 8-OH-DPAT continued to effectively inhibit lordosis behavior following the 5-HT1A receptor agonist's infusion into the more rostral areas. These findings are discussed in relation to earlier studies in which the potency, but not the efficacy, of 8-OH-DPAT was reduced following systemic treatment with the 5-HT1A receptor agonist. PMID- 10373335 TI - Appetitive and consummatory sexual behaviors of female rats in bilevel chambers. I. A correlational and factor analysis and the effects of ovarian hormones. AB - This study investigated measures of sexual behavior displayed by female rats in bilevel chambers, the statistical relationships among the measures, and their dependency on hormone priming. Normative data from a standard 35-min test of sexual behavior were gathered from 82 fully primed sexually experienced Long Evans females and subjected to multiple correlational and factor analyses. Several consummatory measures of copulation were related significantly, whereas appetitive level changing was statistically independent of consummatory measures. Factor analyses were conducted using orthogonal rotations of correlational matrices derived either from (a) measures of female behavior alone or (b) measures of female and male behavior together. The first analysis revealed five factors that accounted for 84% of the intersubject variance: Receptivity, Pacing, Appetitive Level Changing, Lordosis Reflex, and Solicitation. The second factor analysis with male data included revealed seven factors that accounted for 95% of the intersubject variance: Pacing, Copulatory Rate, Mount Count, Receptivity, Appetitive Level Changing, Solicitation, and Lordosis Reflex. Subsequently, subsets of these females were maintained on different steroid priming regimens (oil, low estrogen, high estrogen, high estrogen and progesterone) prior to a standard test of sexual behavior. Although the expression of all sexual behaviors required estrogen priming, appetitive level changing, solicitation, and pacing required progesterone for their full expression. Finally, appetitive level changing developed following hormone treatment alone, regardless of whether the females received access to sexually active males, inactive castrated males, or other females. Use of bilevel chambers allows complex patterns of sexual behavior to be observed in female rats and may thus facilitate the identification of neurochemical or endocrine mechanisms associated with different aspects of female sexual motivation and performance. PMID- 10373336 TI - The roles of stimuli from young, previous breeding experience, and prolactin in regulating parental behavior in ring doves (Streptopelia risoria). AB - In addition to stimulating crop "milk" formation in ring doves, prolactin (PRL) may promote the parental regurgitation behavior that transfers this "milk" to the young at the time of hatching. Although earlier studies suggest that previous breeding experience is an important modulator of PRL-induced parental regurgitation behavior in ring doves, the ways in which experience, hormones, and stimuli from young interact to promote parental behavior have not been well characterized in this species. In the first study, untreated, nonbreeding female doves with and without previous breeding experience were given 10 daily parental behavior tests (2 h/day) with a hungry 5- to 10-day-old foster squab. Experienced females exhibited a higher incidence of regurgitation behavior, defensive behavior, and crouching or sitting in the nest than did inexperienced females. In a second study, nonbreeding females were given 10 daily tests for parental behavior while they received sc injections of ovine PRL or vehicle. Prolactin reduced squab-directed aggression and increased the incidence of regurgitation feeding behavior of foster squabs in both experienced and inexperienced females. However, the average number of regurgitation feeding acts displayed by those PRL treated females that showed the behavior was over eight times higher in experienced females than in inexperienced females. Previous experience also enhanced the stimulatory effects of PRL on defensive behavior and crouching or sitting in the nest. The parental behavior exhibited by nonbreeding, PRL-treated experienced females was qualitatively and quantitatively similar to that observed in normally breeding females during a single test with their own hungry 5- to 10 day-old squabs. These findings indicate that PRL and previous breeding experience both enhance the parental responsiveness of nonbreeding female doves and that under optimal hormonal, experiential, and squab exposure conditions, nonbreeding doves exhibit levels of parental activity that rival those of normally breeding parents. PMID- 10373337 TI - Relationships between dehydroepiandrosterone sulfate (DHEAS) and cortisol (CRT) plasma levels and everyday memory in Alzheimer's disease patients compared to healthy controls. AB - Fifty-two age-matched Alzheimer's disease (AD) patients (26 men, 26 women), mean age 76.2 years, were assessed with the Rivermead Behavioural Memory Test, a test of everyday memory, coincident with the measurement of plasma cortisol (CRT) and dehydroepiandrosterone sulfate (DHEAS) via radioimmunoassay. The AD patients were compared to a control group of age- and gender-matched healthy elderly men and women. No differences were found between the AD patients and the controls in DHEAS or CRT levels, or in the DHEAS/CRT ratio. There were no gender differences in DHEAS or CRT levels, or in the DHEAS/CRT ratio in subjects with AD. However, AD patients with higher levels of DHEAS scored better than those with lower levels on the subtests of Remembering a Name associated with a picture, Digit Span Total and Forward, and the Mini Mental Status Exam. AD patients with higher CRT levels performed worse on Delayed Route Recall than those with lower levels. These findings suggest that AD patients with higher endogenous levels of DHEAS may perform better on some memory tasks than those with lower levels, while AD patients with lower levels of CRT may perform better than those with higher CRT. PMID- 10373338 TI - Regulation of noncontact erection in rats by gonadal steroids. AB - Male rats exhibit erections in the presence of inaccessible estrous females, and we investigated which gonadal steroids regulate these noncontact erections (NCEs). Sexually experienced Wistar males (n >/= 8/group) were tested for NCE four times (every 3 days) before castration, after castration, and after receiving subcutaneous implants of 10-mm Silastic capsules that were empty or filled with crystalline testosterone propionate (TP), dihydrotestosterone (DHT), estradiol benzoate (EB), or DHT + EB (10 mm each). Before castration, males responded with NCE in approximately 50% of tests. No males had NCEs after castration, beginning 3 days after surgery. Also, no males responded after treatment with EB or empty capsules. After receiving implants of TP, DHT, or DHT + EB, 50% of males had NCEs, beginning with the first test 3 days after treatment. On every measure of NCE, males treated with DHT or DHT + EB were indistinguishable from each other and from TP-treated males. Among the sexual responses of male rats, NCE appears to be more sensitive than other behaviors to changes in gonadal condition. In its profile of response to gonadal steroids (testosterone+, dihydrotestosterone+, estradiol-), NCE is similar to reflexive erection, for which spinal systems are sufficient, and unlike copulation (T+, DHT , E+), which depends on discrete areas of the brain. We nonetheless conclude that NCE depends on androgen-sensitive systems in the brain, but androgen-sensitive neurons in the lumbosacral spinal cord may also play a role. PMID- 10373339 TI - Facilitatory and inhibitory effects of beta-endorphin on lordosis in female rats: relation to time of administration. AB - The purpose of the present study was to investigate the effect of time of beta endorphin (beta-EP) administration on lordosis in ovariectomized female rats injected subcutaneously (sc) with estradiol benzoate (EB) and progesterone (Prog). Intracerebroventricular (icv) injections of beta-EP and naloxone (NLX), an opioid receptor antagonist, were administered at the various stages of sc steroid hormone priming. Facilitation of lordosis induced by 10 microg beta-EP was observed exclusively within the initial 6 h of estrogen action, after which inhibition of lordosis occurred. At 12 h after EB priming, at the time of sc Prog treatment (or 43 h after EB priming), icv injection of 10 microg beta-EP significantly inhibited lordosis. Lordosis was significantly facilitated by icv injections of 1 and 10 microg beta-EP at the time of sc EB priming, but not by 0.1 microg beta-EP. A dose-response relationship was identified for lordosis in experimental animals receiving icv injection of beta-EP. Lordosis was inhibited by icv injections of 1 and 10 microg beta-EP at 1 h before the test (or 47 h after EB priming). Lordosis was significantly inhibited by icv injection of NLX at all stages. From the present results, it seems that two different mechanisms are involved in endorphinergic modulation of rats' sexual receptivity: (a) the endorphinergic system at the initial stages of estrogen action facilitates the estrogen activation of lordosis; (b) the endorphinergic system at the final stages of steroid action inhibits lordosis. Moreover, there exists a critical time between 6 and 12 h after estrogen priming for endorphinergic mediation to modulate estrogen action. PMID- 10373340 TI - Although both rhGh and rhIGF-I have the potential to provide clinical benefit, each also has the potential to produce unwanted side-effects. PMID- 10373341 TI - Insulin-like growth factor binding protein (IGFBP) proteolysis: occurrence, identification, role and regulation. PMID- 10373342 TI - The effect of the deterioration of insulin sensitivity on beta-cell function in growth-hormone-deficient adults following 4-month growth hormone replacement therapy. AB - The purpose of the present study was to evaluate the combined effect of GH treatment on body composition and glucose metabolism, with special focus on beta cell function in adult GHD patients. In a double-blind placebo-controlled design, 24 GHD adults (18M/6F), were randomized to 4 months treatment with biosynthetic GH 2 IU/m2s.c. daily (n =13) or placebo (n =11). At inclusion and 4 months later an oral glucose tolerance test (OGTT), a frequently sampled intravenous glucose tolerance test (FSIGT) and dual-energy X-ray absorptiometry (DXA) whole-body scanning were performed. During the study period, body weight decreased 1.6 kg from 94.0 +/- 18.7 to 92.4 +/- 19.4 kg (mean +/- SD) (P<0.05) in the GH-treated group, but remained unchanged in the placebo group. Fat mass decreased from 32.4 +/- 9.6 to 28.1 +/- 10.5 kg (P<0.001), whereas lean body mass increased from 58.3 +/- 11.5 to 61.0 +/- 11.7 kg (P<0.01) in the GH-treated group. Treatment with GH for 4 months resulted in a significant increase in fasting blood glucose (before GH 5.0 +/- 0.3 and after 5.4 +/- 0.6 mmol/l, P<0.05), fasting plasma insulin (before GH 38.4 +/- 30.2 and after 55.3 +/- 34.7 pmol/l, P<0.02) and fasting proinsulin (before 8. 1 +/- 6.7 and after 14.6 +/- 16.1 pmol/l, P<0.05). The insulin sensitivity index SI, estimated by Bergmans Minimal Model, decreased significantly [before GH 1.1 +/- 0.7 and after 0.4 +/- 0.2 10(-4)(min x pmol/l), P<0.003]. The non-insulin-dependent glucose uptake (glucose effectiveness SG did not change (before GH 0.017 +/- 0.005 and after 0.015 +/- 0.006 min-1, NS). Insulin secretion was enhanced during GH therapy, but insufficiently to match the changes in SI, resulting in a higher blood glucose level during an OGTT. Blood glucose at 120 min was 5.5 and 6.3 mmol/l before and after GH treatment, respectively (P = 0.07). One patient developed impaired glucose tolerance. Short term GH replacement therapy in a dose of about 2 IU/m2 daily in GHD adults induces a reduction in insulin sensitivity, despite favourable changes in body composition, and an inadequate enhancement of insulin secretion. PMID- 10373343 TI - Ipamorelin, a new growth-hormone-releasing peptide, induces longitudinal bone growth in rats. AB - Ipamorelin is a new and potent synthetic pentapeptide which has distinct and specific growth hormone (GH)-releasing properties. With the objective of investigating the effects on longitudinal bone growth rate (LGR), body weight (BW), and GH release, ipamorelin in different doses (0, 18, 90 and 450 microg/day) was injected s.c. three times daily for 15 days to adult female rats. After intravital tetracycline labelling on days 0, 6, and 13, LGR was determined by measuring the distance between the respective fluorescent bands in the proximal tibia metaphysis. Ipamorelin dose-dependently increased LGR from 42 microm/day in the vehicle group to 44, 50, and 52 microm/day in the treatment groups (P<0.0001). There was also a pronounced and dose-dependent effect on BW gain. The treatment did not affect total IGF-I levels, IGFBPs, or serum markers of bone formation and resorption. The number of tartrate-resistant acid phosphatase-positive multinuclear cells in the metaphysis of the tibia did not change significantly with treatment. The responsiveness of the pituitary to a provocative i.v. dose of ipamorelin or GHRH showed that the plasma GH response was marginally reduced (P<0.03) after ipamorelin, but unchanged after GHRH. The pituitary GH content was unchanged by ipamorelin treatment. Whether ipamorelin or other GH secretagogues may have a place in the treatment of children with growth retardation requires demonstration in future clinical studies. PMID- 10373344 TI - Regular fluctuations in growth hormone (GH) release determine normal human growth. AB - Growth hormone (GH) is the principal hormone associated with growth through childhood, but in a normal child the amount of GH secretion does not appear to be critical in the generation of normal growth rates. We have assessed the relationship between growth and urinary GH (uGH) output in a longitudinal study of 29 healthy prepubertal schoolchildren (13 male, 16 female; age 5.7-7.8 years) over 1 year. Height and uGH were measured three times a week. Individual height velocity curves were derived using non-linear regression. Growth was expressed in terms of the total increment over the year (DeltaHt, cm), height velocity standard deviation score (HVSDS) and the average size of individual growth spurts. Urinary GH data (ng) were expressed as a weekly average. Mean uGH did not correlate with stature or growth over the year. However, the coefficient of variation of uGH was correlated with height standard deviation score (HtSDS, r = 0.38, P< 0.05), while the relative constancy of short-term change in uGH (coefficient of incremental change, DeltaINC) was inversely correlated with DeltaHt (r = - 0.44) and HVSDS (r = - 0.42, both P< 0.05) but not with HtSDS. DeltaINC was also inversely correlated with the average size of individual growth spurts derived from the height velocity curves (r = - 0.45, P< 0.05). Using time series analysis to identify rhythms in uGH excretion, a positive correlation was found between the magnitude of rhythms of a period of 2 to 4 weeks and HtSDS (r = 0.40, P< 0.05). These data demonstrate that variability in GH is a more important determinant of normal childhood growth rate than the amount of GH alone. Stature is correlated to the overall variability in GH release, while increment in height and the magnitude of individual growth spurts are influenced by the constancy of the GH profile. This would imply that once the GH dose has been replaced in GH deficiency, optimal growth could only be achieved by varying the pattern of GH administration. PMID- 10373345 TI - The effect of growth hormone on rat myocardial collagen. AB - Growth hormone (GH) can increase cardiac performance, but conditions with GH excess, such as acromegaly, are associated with hypertrophy and fibrosis of the heart. The aim of this study was to examine the effect of GH administration on rat myocardial collagen. Female rats were injected with GH (5 mg/kg/day) for 80 days. The weight of the right ventricle (RV) and the left ventricle (LV) was increased in the GH-treated group compared with the control group (P< 0.001). No differences in the ratio of heart weight/body weight or ventricle weight/body weight were found. The total amount of RV and LV collagen was increased in the GH treated group (P< 0.001), but the collagen concentration was decreased (P< 0.001). Histomorphometry showed that the area fraction of collagen relative to myocytes remained unchanged. The composition of ventricular collagen in the GH injected group did not differ from that of the control group concerning the relative amounts of collagen types I and III and pyridinoline, a mature collagen cross-link. We conclude that GH induced a substantial, but proportionate growth of the myocardium without formation of fibrosis. GH actually decreased the collagen concentration, and did not change the composition of myocardial collagen. PMID- 10373346 TI - Increased, not decreased activation of the insulin-like growth factor (IGF) receptor signalling pathway during ceramide-induced apoptosis. AB - The insulin-like growth factors (IGFs) are capable of blocking apoptosis in many cell lines in vitro. The IGF-I receptor (IGF-IR) is believed to mediate protective effects of the IGFs against apoptosis. To determine whether ceramide mediated induction of apoptosis involved a decreased survival effect of the IGF IR, apoptosis was induced in IGF-I receptor positive (R+) and negative (R-) murine fibroblasts by incubation with increasing doses of the sphingolipid analogue, C2 ceramide. Lower ceramide doses were required to induce death in receptor negative compared with receptor positive fibroblasts (P< 0.05 at ceramide doses of 2 microM or greater), not only corroborating evidence that the IGF-I receptor functions as a survival receptor, but also suggesting that ceramide is not inducing apoptosis by suppressing a survival effect of the IGF IR. Ceramide has been reported to induce death through suppression of MAP kinase, and activation of JUN kinase signalling; since our initial data suggested that ceramide had not affected an anti-apoptotic signalling event of the IGF-IR, we monitored the activation of these enzymes. To our surprise, in the presence of ceramide, not only was JUN kinase activity increased, but so too was MAP kinase. Inhibition of MAP kinase, using the MEKK inhibitor, PD98059, significantly reduced ceramide-induced cell death (P< 0. 001). Ceramide also enhanced IGF induced tyrosine phosphorylation of the IGF-I receptor and activated PI-3 kinase. The cumulative effects of these events resulted in increased progression to the G2 phase of the cell cycle, arrest without subsequent mitosis, and apoptosis. These results indicate that ceramide is capable of eliciting apparently contradictory events within a single cell type, and suggest that in the presence of an IGF-IR, survival is enhanced because ceramide can activate PI-3 kinase, believed to be an anti-apoptotic enzyme. PMID- 10373347 TI - Study of growth hormone secretion and action in growth-retarded children with juvenile chronic arthritis (JCA). AB - The stimulated and spontaneous growth hormone (GH) secretion and the response to GH action were assessed in growth-retarded children with juvenile chronic arthritis (JCA), in order to determine the underlying mechanisms of growth retardation in such children. Six children (4 boys and 2 girls aged 10.7-13.8 years) with active JCA of systemic onset were included in the study which involved: (1) anthropometric measurements; (2) assessment of GH responses to insulin-induced hypoglycaemia and clonidine stimulation; (3) assessment of the nocturnal pulsatile GH secretion by measuring GH in blood samples obtained every 20 min from 20.00 to 08.00 h; and (4) the IGF-I generation test. As a control, the latter test was also performed in eight aged-matched children with physiological delay in puberty. Biosynthetic hGH (0.1 IU/kg BW) was administered s. c. for 4 days and blood samples were taken at baseline and the morning after the last GH injection for measurement of IGF-I and IGFBP-3. All six children with JCA were prepubertal and their growth velocity was <3 cm/year. The GH responses to both stimulation tests were normal (peak GH >20 mU/l). Analysis of the pulsatile GH secretion during the night revealed three-to-four GH pulses of normal amplitude (>20 mU/l). IGF-I (26.7+/-4.6 nmol/l, mean+/-SD) and IGFBP-3 (2.1+/-0.2 mg/l) levels were lower in the patients compared with the controls (43.0+/-3.7 nmol/l and 2.8+/-0.2 mg/l, respectively, P<0.01). Following stimulation with exogenous hGH, there was a significant increase in IGF-I and IGFBP-3 levels in the control group (85 and 73%, respectively), but only a small increase in the patients (31 and 14%). It appears that stimulated and spontaneous GH secretion is normal in children with active systemic JCA, but the response to endogenous and exogenous GH with regard to IGF-I and IGFBP-3 production is impaired, indicating a degree of GH insensitivity in such children. PMID- 10373349 TI - Dispersal and metapopulation viability in a heterogeneous landscape AB - Conditions of persistence or extinction of a metapopulation of a colonizing annual species are studied in a heterogeneous landscape, a mixture of two elementary landscapes. An elementary landscape is a landscape whose age-structure is described by only one transition matrix, giving the probability for a site to be disturbed, or to follow the process of succession. We first provide an analytical study of the range of dispersal rates that allow metapopulation persistence in an elementary landscape. Second, conditions for metapopulation persistence in a heterogeneous landscape are derived from results obtained in each elementary landscape. Three cases are distinguished. If the two ranges of dispersal rates defined in each elementary landscape overlap, the metapopulation persists in any mixture of the elementary landscapes. If these two dispersal rates ranges are non-overlapping, either the metapopulation goes extinct for some values of the proportion of the elementary landscapes, or two discontinuous ranges of dispersal rates allow the metapopulation persistence. The consequences of these results are discussed in terms of landscape management. In particular, it is shown that under some conditions, a rapid change in environment (from one elementary landscape to another one) might less often lead to metapopulation extinction than a slower change. Copyright 1999 Academic Press. PMID- 10373348 TI - The pharmacokinetics of free insulin-like growth factor-I in healthy subjects. AB - In a randomized cross-over study in five healthy males we compared 75-min constant i.v. infusion of saline, low-dose recombinant human (rh) insulin-like growth factor-I (rhIGF-I; 1.5 microg/kg/h) and high-dose rhIGF-I (9.0 microg/kg/h). Serum samples were analysed for ultrafiltered free IGF-I (fIGF-I), total IGF-I (tIGF-I), tIGF-II and IGF-binding protein-1 (IGFBP-1) and -3. Free and total IGF-I were unchanged during saline infusion. Low-dose rhIGF-I caused a small increment in fIGF-I [+41%, from 0.64 +/- 0.19 (mean +/- SEM) to 0.90 +/- 0.25 microg/l;P< 0.05] and tIGF-I (+9%, from 220 +/- 31 to 239 +/- 33 microg/l;P< 0.05). High-dose rhIGF-I increased tIGF-I by 40% (from 227 +/- 36 to 329 +/- 31 microg/l;P< 0.05), and fIGF-I by 11.5 times (from 0.56 +/- 0.20 to 6.46 +/- 1.39 microg/l;P< 0.05). The pharmacokinetic profile of fIGF-I was calculated after high-dose IGF-I only. The disappearance of fIGF-I followed first order kinetics with an apparent half-life of 14.4 +/- 1.0 [11.2-17.1 (range)] min. The clearance was estimated to 52 +/- 20 (16-128) ml/min/kg and the volume of distribution to 1102 +/- 464 (388-2899) ml/kg. In the three experiments, there were no differences in IGFBP-1, and tIGF-II and IGFBP-3 remained unchanged. In conclusion, fIGF-I remained within the physiological range after low-dose rhIGF I, whereas high-dose rhIGF-I resulted in supraphysiological concentrations. Since the half-life estimates for each subject were remarkably similar, this parameter most likely does not explain the observed variation in clearance and volume of distribution of fIGF-I. Instead, differences in the circulating and cellular IGF I binding capacity may be of importance. PMID- 10373350 TI - A dynamic model of the meningococcal transferrin receptor. AB - Iron is an essential nutrient for all organisms and consequently, the ability to bind transferrin and sequester iron from his source constitutes a distinct advantage to a blood-borne bacterial pathogen. Levels of free iron are strictly limited in human serum, largely through the action of the iron-binding protein transferrin. The acquisition of trasferrin-iron is coincident with pathogenicity among Neisseria species and a limited number of other pathogens of human and veterinary significance. In Neisseria meningitidis, transferrin binding relies on two co-expressed, outer membrane proteins distinct in aspects of both structure and function. These proteins are independently and simultaneously capable of binding human transferrin and both are required for the optimal uptake of iron from this source. It has been established that transferrin-binding proteins (designated TbpA and TbpB) form a discrete, specific complex which may be composed of a transmembrane species (composed of the TbpA dimer) associated with a single surface-exposed lipoprotein (TbpB). This more exposed protein is capable of selectively binding iron-saturated transferrin and the receptor complex has ligand-binding properties which are distinct from either of its components. Previous in vivo analyses of N. gonorrhoeae, which utilizes a closely related transferrin-iron uptake system, indicated that this receptor exists in several conformations influenced in part by the presence (or absence) of transferrin. Here we propose a dynamic model of the meningococcal transferrin receptor which is fully consistent with the current data concerning this subject. We suggest that TbpB serves as the initial binding site for iron-saturated transferrin and brings this ligand close to the associated transmembrane dimer, enabling additional binding events and orientating transferrin over the dual TbpA pores. The antagonistic association of these receptor proteins with a single ligand molecule may also induce conformational change in transferrin, thereby favouring the release of iron. As, in vivo, transferrin may have iron in one or both lobes, this dynamic molecular arrangement would enable iron uptake from either iron binding site. In addition, the predicted molecular dimensions of the putative TbpA dimer and hTf are fully consistent with these proposals. Given the diverse data used in the formulation of this model and the consistent characteristics of transferrin binding among several significant Gram-negative pathogens, we speculate that such receptor-ligand interactions may be, at least in part, conserved between species. Consequently, this model may be applicable to bacteria other than N. meningitidis. PMID- 10373351 TI - Collective decision-making in social spiders: dragline-mediated amplification process acts as a recruitment mechanism AB - Amplification mechanisms involved in group cohesion and coordination of several individuals' activities are a major research topic in social arthropod biology. In this paper, we investigate how recruitment processes can be mediated by the use of silk draglines in the case of social spiders. Our intent is to demonstrate how a behavioural feature common to all spider species can induce positive feedback, potentially leading to collective decision-making in a social context. Dragline-mediated amplification mechanisms are investigated in a simulated "Y" choice set-up. Numerical experiments involve two distinct models: a simplified one, devoted to the exploration of the most basic amplifying properties of the system, and a more complex simulation platform, taking into account the geometrical properties of a growing network (two-dimensional web). The effect of hypothetical subpopulations, as well as variations in silk attractivity in the case of mixed populations (originating from multiple nests) are also discussed. Results fit experimental data and demonstrate that spiders' behaviour exhibits very strong amplifying properties that can play a crucial part in the organization of social life. Copyright 1999 Academic Press. PMID- 10373352 TI - The evolutionary ecology of dominance-recessivity. AB - An "adaptive dynamics" modelling approach to the evolution of dominance recessivity is presented. In this approach, fitness derives from an explicit ecological scenario, and both evolutionary attractivity and invasibility of resident populations are examined. The ecology consists of a within-individual part representing a locus with regulated activity and a between-individual part that is a two-patch soft selection model. Evolutionary freedom is allowed at a single locus. The evolutionary analysis considers directed random walks on trait space, generated by repeated invasions of mutants. The phenotype of an individual is determined by allelic parameters. Mutations can have two effects: they either affect the affinity of the promoter sequence for transcription factors, or they affect the gene product. The dominance interaction between alleles derives from their promoter affinities. Additive genetics is evolutionarily unstable when selection and evolution maintain two alleles in the population. In such a situation, dominance interactions can become stationary and close to additive genetics or they continue to evolve at a very slow pace towards dominance recessivity. The probability that a specific dominance interaction will evolve depends on the relative mutation rate of promoter compared to gene product and the distribution of mutational effect sizes. Either allele in the dimorphism can become dominant, and dominance-recessivity is always most likely to evolve. Evolution then approaches a population state where every phenotype has maximum viability in one of the two patches. When the within-individual part is replaced by a housekeeping locus that codes for a metabolic enzyme, evolution favours a population of two alleles under the same conditions as for a regulated locus. In the case of a housekeeping gene, however, the evolutionary dynamical system approaches a population state where the heterozygote and only one homozygote phenotype are equivalent to the optimum phenotypes in the two patches. PMID- 10373353 TI - Animal group forces resulting from predator avoidance and competition minimization AB - A new model to explain animal spacing, based on a trade-off between foraging efficiency and predation risk, is derived from biological principles. The model is able to explain not only the general tendency for animal groups to form, but some of the attributes of real groups. These include the independence of mean animal spacing from group population, the observed variation of animal spacing with resource availability and also with the probability of predation, and the decline in group stability with group size. The appearance of "neutral zones" within which animals are not motivated to adjust their relative positions is also explained. The model assumes that animals try to minimize a cost potential combining the loss of intake rate due to foraging interference and the risk from exposure to predators. The cost potential describes a hypothetical field giving rise to apparent attractive and repulsive forces between animals. Biologically based functions are given for the decline in interference cost and increase in the cost of predation risk with increasing animal separation. Predation risk is calculated from the probabilities of predator attack and predator detection as they vary with distance. Using example functions for these probabilities and foraging interference, we calculate the minimum cost potential for regular lattice arrangements of animals before generalizing to finite-sized groups and random arrangements of animals, showing optimal geometries in each case and describing how potentials vary with animal spacing. Copyright 1999 Academic Press. PMID- 10373354 TI - The blood-stage dynamics of mixed Plasmodium malariae-Plasmodium falciparum infections. AB - We present the first mathematical model of the within-host dynamics of a mixed species malaria infection in a human: the blood-stage population dynamics of a dual infection with Plasmodium malariae and Plasmodium falciparum. Our results reproduce several important features of such infections in nature, including the asymmetry of species asexual-form densities, inter-specific suppression through interactions with the human immune system, and seasonal alternations in species prevalence. Most importantly, our results suggest that an existing P. malariae infection can reduce the peak parasitemia of a subsequent P. falciparum superinfection by as much as 50%. This result integrates numerous empirical observations and supports the hypothesis that clinical outcomes of P. falciparum infections may be influenced by the presence of a congener. PMID- 10373355 TI - Estimating temporal independence of radio-telemetry data on animal activity AB - Radio-telemetry is an excellent tool for gathering data on the biology of animals and their interactions with the environment they inhabit. Many methods have been developed for analyses of spatial information, on home range size and utilization density. Activity patterns are often described using radio-tracking data, but no generally accepted method is currently available specifically for determining the temporal independence of this type of data for statistical inference. Activity rhythms have generally been analysed by ecologists with the assumption that data are temporally independent, or by subjectively fixing an independence interval, based on attributes of their ranging behaviour. Although some good approximations of activity patterns can be obtained in these ways, we underline the need for a functionally correct method of estimating independence interval. Here we use semi variograms to estimate the minimum interval required for the readings to be sequentially independent. This geostatistical tool is applied to the analysis of data on activity of Chilean foxes (Pseudalopex culpaeus) and Chacoan peccaries (Catagonus wagneri). Data were collected in the field by radio-tracking over 24 hr periods, with readings on activity state taken every 15 min. The spatial dimension in which the theory of geostatistics lies has been transferred into the time dimension, so that the correlation interval is expressed in time units (min). Time of independence as estimated by the variogram was 110 min for foxes, while data on peccaries indicated that they have long periods of activity, more suitable for time-series analysis. Copyright 1999 Academic Press. PMID- 10373356 TI - Emerging patterns and food recruitment in ants: an analytical study AB - A model of food recruitment by social insects accounting for the competition between trails in the presence of an arbitrary number of sources is developed and analysed in detail. Both the case of identical environmental characteristics and the case where one source and the corresponding trail are different from the others are considered. Different collective responses depending on the environmental conditions, and without change of individual behaviour, are shown to exist, associated with the possibility that the colony may be led to exploit one source or a group of sources preferentially. The full bifurcation diagram of steady-state solutions is constructed from which the dominant exploitation patterns are identified. The biological relevance of the results is discussed and suggestions are made for their experimental testing in connection with the recruitment behavior of species using trail recruitment. The same phenomenological model can be used for different trail-laying species since the predictions are generic and not restricted to a given species, except for the parameter values used. Copyright 1999 Academic Press. PMID- 10373357 TI - The validity of the handicap principle in discrete action-response games AB - The validity of the handicap principle has spawned much debate in spite of the existence of a formal treatment. Simple models constructed to further investigate the issue were able both to prove and to disprove some of its claims. Here I show with the aid of a more general model, which takes into account both assumptions presented in these previous simple models: (1) that the previous results are not in conflict since they can be obtained as specific cases of this general model; (2) that ESS communication need not use costly signals, that is, even under conflict of interest, the cost of a signal used by a high-quality individual can be zero (or even negative) provided that the cost for low-quality signallers is high enough; (3) that only the cost relative to the benefits of the interaction should be higher for worse signallers; and (4) that in a discrete model the differential cost is only a necessary but not a sufficient condition for evolutionarily stable reliable communication. Copyright 1999 Academic Press. PMID- 10373358 TI - The X-ray crystal structure of a Valpha2.6Jalpha38 mouse T cell receptor domain at 2.5 A resolution: alternate modes of dimerization and crystal packing. AB - We describe here the structure of a murine T cell receptor (TCR) Valpha2.6Jalpha38 (TCRAV2S6J38) domain, derived from a T cell hybridoma with specificity for the H-2Ddmajor histocompatibility complex class I molecule bound to a decamer peptide, P18-I10, from the HIV envelope glycoprotein gp120, determined by X-ray crystallography at 2.5 A resolution. Unlike other TCR Valpha domains that have been studied in isolation, this one does not dimerize in solution at concentrations below 1 mM, and the crystal fails to show dimer contacts that are likely to be physiological. In comparison to other Valpha domains, this Valpha2.6 shows great similarity in the packing of its core residues, and exhibits the same immunoglobulin-like fold characteristic of other TCR Valpha domains. There is good electron density in all three complementarity determining regions (CDRs), where the differences between this Valpha domain and others are most pronounced, in particular in CDR3. Examination of crystal contacts reveals an association of Valpha domains distinct from those previously seen. Comparison with other Valpha domain structures reveals variability in all loop regions, as well as in the first beta strand where placement and configuration of a proline residue at position 6, 7, 8, or 9 affects the backbone structure. The great variation in CDR3 conformations among TCR structures is consistent with an evolving view that CDR3 of TCR plays a plastic role in the interaction of the TCR with the MHC/peptide complex as well as with CDR3 of the paired TCR chain. PMID- 10373360 TI - Determinants of the stability of transcription elongation complexes: interactions of the nascent RNA with the DNA template and the RNA polymerase. AB - We use a synthetic "primed bubble-duplex" model elongation complex developed previously to examine certain structural and thermodynamic features of the transcription elongation complex of Escherichia coli. The nucleic acid framework of this model complex consists of a linear base-paired DNA molecule with a central "bubble" of non-complementary nucleotide residues, together with a single stranded RNA molecule that is complementary (at its 3'-end) to three to 12 nucleotide residues of one of the DNA strands within the bubble. RNA polymerase is added to this framework in trans, and on addition of rNTPs the resulting complex can elongate the 3'-end of the RNA primer in a template-dependent manner with functional properties that are indistinguishable from those of a "natural" promoter-initiated transcription elongation complex operating under the same conditions. In this study we use this model system to show that the formation of a stable elongation complex at any particular template position can be treated as an equilibrium process, and that semi-quantitative dissociation constants can be estimated for the complex by using a gel band-shift assay to monitor the binding of the RNA oligomer to the complex. We then show that the formation of a stable complex depends on the presence of a complementary RNA-DNA hybrid that is at least 9 bp in length, and in addition that several nucleotide residues of non complementary RNA located upstream of the RNA-DNA hybrid bind strongly to the putative single-stranded RNA binding site of the polymerase and significantly enhance the stability of the resulting elongation complex. Finally, we demonstrate that the measured stabilities of the model constructs in which the length of the RNA-DNA hybrid is varied correlate well with the transcriptional processivity of the functioning complex that results when rNTPs are added. These findings are discussed in the context of related studies of both model systems and natural elongation complexes. The general concepts that emerge are used to define some central structural and functional features of the transcription complex. PMID- 10373359 TI - Analysis of 27 mammalian and 9 avian PrPs reveals high conservation of flexible regions of the prion protein. AB - Prion diseases are fatal neurodegenerative disorders in man and animal associated with conformational conversion of a cellular prion protein (PrPc) into the pathologic isoform (PrPSc). The function of PrPcand the tertiary structure of PrPScare unclear. Various data indicate which parts of PrP might control the species barrier in prion diseases and the binding of putative factors to PrP. To elucidate these features, we analyzed the evolutionary conservation of the prion protein. Here, we add the primary PrP structures of 20 ungulates, three rodents, three carnivores, one maritime mammal, and nine birds. Within mammals and birds we found a high level of amino acid sequence identity, whereas between birds and mammals the overall homology was low. Various structural elements were conserved between mammals and birds. Using the CONRAD space-scale alignment, which predicts conserved and variable blocks, we observed similar patterns in avian and mammalian PrPs, although 130 million years of separate evolution lie in between. Our data support the suggestion that the repeat elements might have expanded differently within the various classes of vertebrates. Of note is the N-terminal part of PrP (amino acid residues 23-90), which harbors insertions and deletions, whereas in the C-terminal portion (91-231) mainly point mutations are found. Strikingly, we found a high level of conservation of sequences that are not part of the structured segment 121-231 of PrPcand of the structural elements therein, e.g. the N-terminal region from amino acid residue 23-90 and the regions located upstream of alpha-helices 1 and 3. PMID- 10373361 TI - All three residues of the Tn 10 transposase DDE catalytic triad function in divalent metal ion binding. AB - Tn 10/IS 10 transposition involves excision of the transposon from a donor site and subsequent joining of the excised transposon to a new target site. These steps are catalyzed by the Tn 10 -encoded transposase protein and require the presence of a suitable divalent metal ion. Like other transposase and retroviral integrase proteins, Tn 10 transposase appears to contain a single active site which includes a triad of acidic amino acid residues generally referred to as the DDE motif. In addition to its role in catalysis, the Tn 10 transposase DDE motif also functions in target capture, a step that in vitro is greatly facilitated by the presence of a suitable divalent metal ion. We show that cysteine residue substitutions at each of the DDE motif residues in Tn 10 transposase result in a change in the divalent metal ion requirements for catalysis, such that Mn2+but not Mg2+can be used. This switch in metal ion specificity provides evidence that each of the DDE motif residues functions directly in metal ion binding. We also show differential effects of DDE mutations on metal ion-assisted target capture. A number of models, including a two metal ion active site, are considered to explain these effects. PMID- 10373362 TI - Analysis of DNA replication forks encountering a pyrimidine dimer in the template to the leading strand. AB - Electron microscopy (EM) was used to visualize intermediates of in vitro replication of closed circular DNA plasmids. Cell-free extracts were prepared from human cells that are proficient (IDH4, HeLa) or deficient (CTag) in bypass replication of pyrimidine dimers. The DNA substrate was either undamaged or contained a single cis, syn thymine dimer. This lesion was inserted 385 bp downstream from the center of the SV40 origin of replication and sited specifically in the template to the leading strand of the newly synthesized DNA. Products from 30 minute reactions were crosslinked with psoralen and UV, linearized with restriction enzymes and spread for EM visualization. Extended single-stranded DNA regions were detected in damaged molecules replicated by either bypass-proficient or deficient extracts. These regions could be coated with Escherichia coli single-stranded DNA binding protein. The length of duplex DNA from a unique restriction site to the single-stranded DNA region was that predicted from blockage of leading strand synthesis by the site-specific dimer. These results were confirmed by S1nuclease treatment of replication products linearized with single cutting restriction enzymes, followed by detection of the diagnostic fragments by gel electrophoresis. The absence of an extended single stranded DNA region in replication forks that were clearly beyond the dimer was taken as evidence of bypass replication. These criteria were fulfilled in 17 % of the molecules replicated by the IDH4 extract. PMID- 10373363 TI - Topological selectivity of a hybrid site-specific recombination system with elements from Tn3 res/resolvase and bacteriophage P1 loxP/Cre. AB - In order to investigate the functions of the parts of the Tn 3 recombination site res, we created hybrid recombination sites by placing the loxP site for Cre recombinase adjacent to the "accessory" resolvase-binding sites II and III of res. The efficiency and product topology of in vitro recombination by Cre between two of these hybrid sites were affected by the addition of Tn 3 resolvase. The effects of resolvase addition were dependent on the relative orientation and spacing of the elements of the hybrid sites. Substrates with sites II and III of res close to loxP gave specific catenated or knotted products (four-noded catenane, three-noded knot) when resolvase and Cre were added together. The product topological complexity increased when the length of the spacer DNA segment between loxP and res site II was increased. Similar resolvase-induced effects on Cre recombination product topology were observed in reactions of substrates with loxP sites adjacent to full res sites. The results demonstrate that the res accessory sites are sufficient to impose topological selectivity on recombination, and imply that intertwining of two sets of accessory sites defines the simple catenane product topology in normal resolvase-mediated recombination. They are also consistent with current models for the mechanism of catalysis by Cre. PMID- 10373364 TI - The RAD51 protein supports homologous recombination by an exchange mechanism in mammalian cells. AB - Information concerning the function of recombination proteins in mammalian cells has been obtained from biochemical studies, but little is known about their mechanisms of action in growing cells. The eukaryotic recombination protein RAD51, a homologue of the Escherichia coli RecA protein, has been shown to interact with various proteins, including the p53 protein, the guardian of genomic stability maintenance. Here, the hamster RAD51 protein, CgRAD51, has been overexpressed in the SPD8 cell line, derived from Chinese hamster V79 cells. This cell line offers unique possibilities for studying different mechanisms for homologous recombination on endogenous substrates. We report that the SPD8 cell line contains a mutated p53 gene, which provides new insights into the recombination process in these cells. The present study demonstrates that overexpression of CgRAD51 in these cells results in a two- to threefold increase in endogenous recombination. In addition, sequence analysis indicated that RAD51 promotes homologous recombination by a chromatid exchange mechanism. PMID- 10373365 TI - Molecular characterization of a protease secreted by Erwinia amylovora. AB - A protease with a molecular mass of 48 kDa is secreted by the fire blight pathogen Erwinia amylovora in minimal medium. We characterized this activity as a metalloprotease, since the enzyme was inhibited by EDTA and o -phenanthroline. A gene cluster was determined to encode four genes connected to protease expression, including a structural gene (prtA) and three genes (prtD, prtE, prtF) for secretion of the protease, which are transcribed in the same direction. The organization of the protease gene cluster in E. amylovora is different from that in other Gram-negative bacteria, such as Erwinia chrysanthemi, Pseudomonas aeruginosa and Serratia marcescens. On the basis of the conservative motif of metalloproteases, PrtA was identified to be a member of the metzincin subfamily of zinc-binding metalloproteases, and was confirmed to be the 48 kDa protease on gels by sequencing of tryptic peptide fragments derived from the protein. The protease is apparently secreted into the external medium through the type I secretion pathway via PrtD, PrtE and PrtF which share more than 90% identity with the secretion apparatus for lipase of S. marcescens. A protease mutant was created by Tn 5 -insertions, and the mutation localized in the prtD gene. The lack of protease reduced colonization of an E. amylovora secretion mutant labelled with the gene for the green fluorescent protein (gfp) in the parenchyma of apple leaves. PMID- 10373366 TI - Filamentous phage are released from the bacterial membrane by a two-step mechanism involving a short C-terminal fragment of pIII. AB - Filamentous phage assemble at the membrane of infected cells. The phage filament is released from the membrane at the end of assembly, after four to five copies of the minor proteins, pIII and pVI, have been added to the end of the virion. In the absence of pIII or pVI, phage filaments are not released, but remain associated with the cells. The C-terminal portion of pIII, termed the "C" domain, is required for the release of stable virions. With the use of pIII C-terminal fragments of increasing size, termination of assembly can be divided into various steps. An 83-residue fragment leads to the incorporation of pVI into the assembling phage, but does not release it from the membrane. A slightly longer fragment (93 residues) is sufficient to release the particle into the culture supernatant. However, these released particles are unstable in the detergent, sarkosyl, which does not disrupt wild-type phage. A fragment of >121 residues is needed for the particle to become detergent resistant. Thus, the C-domain can be divided into two subdomains: C2, sufficient for release, and C1, required for virion stability.A model for termination of phage assembly is proposed in which pIII and pVI dock to the membrane-associated filament and form a pre- termination complex. Then, a conformational change involving the C2 domain of pIII disrupts the hydrophobic interactions with the inner membrane, releasing the phage from the cells. The pIII-mediated release of phage from the membranes points to one possible mechanism for excision of membrane-anchored protein complexes from lipid bilayers. PMID- 10373367 TI - Solution structure and thermodynamics of a divalent metal ion binding site in an RNA pseudoknot. AB - Identification and characterization of a metal ion binding site in an RNA pseudoknot was accomplished using cobalt (III) hexammine, Co(NH3)63+, as a probe for magnesium (II) hexahydrate, Mg(H2O)62+, in nuclear magnetic resonance (NMR) structural studies. The pseudoknot causes efficient -1 ribosomal frameshifting in mouse mammary tumor virus. Divalent metal ions, such as Mg2+, are critical for RNA structure and function; Mg2+preferentially stabilizes the pseudoknot relative to its constituent hairpins. The use of Co(NH3)63+as a substitute for Mg2+was investigated by ultraviolet absorbance melting curves, NMR titrations of the imino protons, and analysis of NMR spectra in the presence of Mg2+or Co (NH3)63+. The structure of the pseudoknot-Co(NH3)63+complex reveals an ion-binding pocket formed by a short, two-nucleotide loop and the major groove of a stem. Co(NH3)63+stabilizes the sharp loop-to-stem turn and reduces the electrostatic repulsion of the phosphates in three proximal strands. Hydrogen bonds are identified between the Co(NH3)63+protons and non-bridging phosphate oxygen atoms, 2' hydroxyl groups, and nitrogen and oxygen acceptors on the bases. The binding site is significantly different from that previously characterized in the major groove surface of tandem G.U base-pairs, but is similar to those observed in crystal structures of a fragment of the 5 S rRNA and the P5c helix of the Tetrahymena thermophila group I intron. Changes in chemical shifts occurred at the same pseudoknot protons on addition of Mg2+as on addition of Co(NH3)63+, indicating that both ions bind at the same site. Ion binding dissociation constants of approximately 0.6 mM and 5 mM (in 200 mM Na+and a temperature of 15 degrees C) were obtained for Co(NH3)63+and Mg2+, respectively, from the change in chemical shift as a function of metal ion concentration. An extensive array of non-sequence-specific hydrogen bond acceptors coupled with conserved structural elements within the binding pocket suggest a general mode of divalent metal ion stabilization of this type of frameshifter pseudoknot. These results provide new thermodynamic and structural insights into the role divalent metal ions play in stabilizing RNA tertiary structural motifs such as pseudoknots. PMID- 10373368 TI - Equilibrium unfolding pathway of an H-type RNA pseudoknot which promotes programmed -1 ribosomal frameshifting. AB - The equilibrium unfolding pathway of a 41-nucleotide frameshifting RNA pseudoknot from the gag-pro junction of mouse intracisternal A-type particles (mIAP), an endogenous retrovirus, has been determined through analysis of dual optical wavelength, equilibrium thermal melting profiles and differential scanning calorimetry. The mIAP pseudoknot is an H-type pseudoknot proposed to have structural features in common with the gag-pro frameshifting pseudoknots from simian retrovirus-1 (SRV-1) and mouse mammary tumor virus (MMTV). In particular, the mIAP pseudoknot is proposed to contain an unpaired adenosine base at the junction of the two helical stems (A15), as well as one in the middle of stem 2 (A35). A mutational analysis of stem 1 hairpins and compensatory base-pair substitutions incorporated into helical stem 2 was used to assign optical melting transitions to molecular unfolding events. The optical melting profile of the wild-type RNA is most simply described by four sequential two-state unfolding transitions. Stem 2 melts first in two closely coupled low-enthalpy transitions at low tmin which the stem 3' to A35, unfolds first, followed by unfolding of the remainder of the helical stem. The third unfolding transition is associated with some type of stacking interactions in the stem 1 hairpin loop not present in the pseudoknot. The fourth transition is assigned to unfolding of stem 1. In all RNAs investigated, DeltaHvH approximately DeltaHcal, suggesting that DeltaCpfor unfolding is small. A35 has the thermodynamic properties expected for an extrahelical, unpaired nucleotide. Deletion of A15 destabilizes the stem 2 unfolding transition in the context of both the wild-type and DeltaA35 mutant RNAs only slightly, by DeltaDeltaG degrees approximately 1 kcal mol-1(at 37 degrees C). The DeltaA15 RNA is considerably more susceptible to thermal denaturation in the presence of moderate urea concentrations than is the wild type RNA, further evidence of a detectable global destabilization of the molecule. Interestingly, substitution of the nine loop 2 nucleotides with uridine residues induces a more pronounced destabilization of the molecule (DeltaDeltaG degrees approximately 2.0 kcal mol-1), a long-range, non-nearest neighbor effect. These findings provide the thermodynamic basis with which to further refine the relationship between efficient ribosomal frameshifting and pseudoknot structure and stability. PMID- 10373370 TI - Intermediate species possessing bent DNA are present along the pathway to formation of a final TBP-TATA complex. AB - Binding of the TATA-binding protein (TBP) to the "TATA" sequences present in the promoters of eukaryotic class II genes is the first step in the sequential assembly of transcription pre-initiation complexes. Myriad structural changes, including severe bending of the DNA, accompany TBP-TATA complex formation. A detailed kinetic study has been conducted to elucidate the mechanistic details of TBP binding and DNA bending. The binding of Saccharomyces cerevisiae TBP to the adenovirus major late promoter (AdMLP) was followed in real-time through a range of temperatures and TBP concentrations using fluorescence resonance energy transfer (FRET) and stopped-flow mixing. The results of association and relaxation kinetics and equilibrium binding experiments were analyzed globally to obtain the complete kinetic and energetic profile of the reaction. This analysis reveals a complex mechanism with two intermediate species, with the DNA in the intermediates apparently bent similarly to the DNA in the final complex. TBP binding and DNA bending occur simultaneously through the multiple steps of the reaction. The first and third steps in this sequential process show nearly identical large increases in both enthalpy and entropy, whereas the middle step is highly exothermic and proceeds with a large decrease in entropy. The first intermediate is significantly populated at equilibrium and resembles the final complex both structurally and energetically. It is postulated that both this intermediate and the final complex bind transcription factor IIB in the second step of pol II pre-initiation complex assembly. A consequence of such a reactive intermediate is that the rate of assembly of transcriptionally competent pre initiation complexes from bi-directionally bound TBP is greatly increased. PMID- 10373371 TI - Three-dimensional reconstruction of Lumbricus terrestris hemoglobin at 22 A resolution: intramolecular localization of the globin and linker chains. AB - A 3D reconstruction of the hemoglobin (Hb) of the earthworm Lumbricus terrestris was carried out by the 3D projection alignment method from electron microscopy images of a frozen-hydrated specimen at 22 A resolution. The results were analyzed by a new approach taking into account the evolution of the 210 densities forming the 3D volume as a function of the threshold of surface representation. The whole oligomer with D6point-group symmetry is comprised of 12 hollow globular substructures (HGS) with local 3-fold symmetry tethered to a complex network of linking subunits (linker complex). The 12 globin subunits of each HGS are distributed around local 3-fold axis in four layers of three subunits. The first layer, the most external, contains monomeric globin chains 2A, 3A, and 5A. The three trimers corresponding to the nine remaining subunits have one subunit in each of the second (2B, 3B, 5B), third (1A, 4A, 6A), and fourth (1B, 4B, 6B) layer. The distances between the centers of the globin chains forming the trimers are in the ranges 20-32 A and 45-52 A. The linker complex is made up of two types of linking units. The first type forms three loops connecting globin chains of the second, third and fourth layers. The average molecular mass (Mm) of these subunits was 25 kDa. The second type forms the central structure, termed hexagonal toroid, and its 12 connections to the HGS. This structure corresponds to a hexamer of a single linking unit with a Mm (31.2 kDa), size and a shape different from those of the HGS loops. A careful study of 3D volume architecture shows that each toroid linking unit is bound to the three loops of a HGS pair located in the upper and lower hexagonal layers, respectively. As shown in a model of architecture, hexagonal bilayered (HBL) Hbs can be built very simply from 144 globin chains and 42 linker chains belonging to two different types. We also propose a simple assembly sequence for the construction of HBL Hbs based on the architecture model. PMID- 10373369 TI - DNA stretching and compression: large-scale simulations of double helical structures. AB - Computer-simulated elongation and compression of A - and B -DNA structures beyond the range of thermal fluctuations provide new insights into high energy "activated" forms of DNA implicated in biochemical processes, such as recombination and transcription. All-atom potential energy studies of regular poly(dG).poly(dC) and poly(dA).poly(dT) double helices, stretched from compressed states of 2.0 A per base-pair step to highly extended forms of 7.0 A per residue, uncover four different hyperfamilies of right-handed structures that differ in mutual base-pair orientation and sugar-phosphate backbone conformation. The optimized structures embrace all currently known right-handed forms of double helical DNA identified in single crystals as well as non-canonical forms, such as the original "Watson-Crick" duplex with trans conformations about the P-O5' and C5'-C4' backbone bonds. The lowest energy minima correspond to canonical A and B form duplexes. The calculations further reveal a number of unusual helical conformations that are energetically disfavored under equilibrium conditions but become favored when DNA is highly stretched or compressed. The variation of potential energy versus stretching provides a detailed picture of dramatic conformational changes that accompany the transitions between various families of double-helical forms. In particular, the interchanges between extended canonical and non-canonical states are reminiscent of the cooperative transitions identified by direct stretching experiments. The large-scale, concerted changes in base-pair inclination, brought about by changes in backbone and glycosyl torsion angles, could easily give rise to the observed sharp increase in force required to stretch single DNA molecules more than 1.6-1.65 times their canonical extension. Our extended duplexes also help to tie together a number of previously known structural features of the RecA-DNA complex and offer a self-consistent stereochemical model for the single-stranded/duplex DNA recognition brought in register by recombination proteins. The compression of model duplexes, by contrast, yields non-canonical structures resembling the deformed steps in crystal complexes of DNA with the TATA-box binding protein (TBP). The crystalline TBP-bound DNA steps follow the calculated compression-elongation pattern of an unusual "vertical" duplex with base planes highly inclined with respect to the helical axis, exposed into the minor groove, and accordingly accessible for recognition.Significantly, the double helix can be stretched by a factor of two and compressed roughly in half before its computed internal energy rises sharply. The energy profiles show that DNA extension-compression is related not only to the variation of base-pair Rise but also to concerted changes of Twist, Roll, and Slide. We suggest that the high energy "activated" forms calculated here are critical for DNA processing, e.g. nucleo-protein recognition, DNA/RNA synthesis, and strand exchange. PMID- 10373372 TI - The role of linkers in the reassembly of the 3.6 MDa hexagonal bilayer hemoglobin from Lumbricus terrestris. AB - The extent and kinetics of reassembly of the four groups of linkers L1-L4 with 213 kDa subassemblies of twelve globin chains D, (bac)3(d)3, isolated from the approximately 3.6 MDa hexagonal bilayer (HBL) hemoglobin (Hb) of Lumbricus terrestris, was investigated using gel filtration. The reassembled HBL's were characterized by scanning transmission electron microscopic (STEM) mass mapping and their subunit content determined by reversed-phase chromatography. In reassembly by method (A), the linkers isolated by RP-HPLC at pH approximately 2.2 were added to D at neutral pH; in method (B), the linkers were renatured at neutral pH and then added to D. With method (A) the percentage of HBL reassembly varied from >/=13% in the absence of Ca(II) to /=75%), with ternary and binary linker combinations (40-50%) and with individual linkers producing yields increasing in the following order: L1=1-3%, L2 approximately L3=10-20% and L4=35-55%. The yield was two- to eightfold lower with method (B), except in the case of linkers L1-L3. Although the reassembly kinetics were always biphasic, with t1/2=0.3-3.3 hours and 10-480 hours, the ratio of the amplitudes fast:slow was 1:0.6 with method (A) and 1:2.5 with method (B). These results are consistent with a scheme in which the slow HBL reassembly is dependent on a slow conversion of linker conformation at neutral pH from a reassembly incompetent to a reassembly competent conformation. Although all the linkers self-associate extensively at neutral pH, forming complexes ranging from dimers to >18-mers, the size of the complex does not affect the extent or rate of reassembly. The oxygen binding affinity of reassembled HBLs was similar to that of the native Hb, but their cooperativity was lower. A model of HBL reassembly was proposed which postulates that binding of linker dimers to two of the three T subunits of D causes conformational alterations resulting in the formation of complementary binding sites which permit lateral self-association of D subassemblies, and thus dictate the formation of a hexagonal structure due to the 3-fold symmetry of D. PMID- 10373373 TI - Crystal structure of a ternary complex between human chorionic gonadotropin (hCG) and two Fv fragments specific for the alpha and beta-subunits. AB - Human chorionic gonadotropin (hCG), is a placental hormone which exerts its major effect by stimulating progesterone production, crucially sustaining the early weeks of pregnancy. Detection of hCG with specific monoclonal antibodies (mAbs) has become the chosen means for pregnancy diagnosis. We have used antibody Fv fragments derived from two high-affinity mAbs, one against the alpha and the other against the beta-hCG subunit to enable the crystallisation of intact or desialylated hCG. Crystals of a ternary complex composed of Fv anti-alpha/hCG/Fv anti-beta were found to diffract to 3.5 A resolution, and the structure was solved by molecular replacement. In the crystal, the two Fvs keep hCG as in a molecular cage, providing good protein-protein contacts and leaving enough space for the saccharides to be accommodated in the cell solvent. The two Fvs were found not to interact directly through their complementary-determining regions with the hCG saccharides, but only with the protein. The hCG structure in the ternary complex was very close to that of the HF partially deglycosylated hormone, thus indicating that neither the saccharides nor the Fvs had any substantial influence on hormone structure. PMID- 10373374 TI - High-resolution solution structure of the 18 kDa substrate-binding domain of the mammalian chaperone protein Hsc70. AB - The three-dimensional structure for the substrate-binding domain of the mammalian chaperone protein Hsc70 of the 70 kDa heat shock class (HSP70) is presented. This domain includes residues 383-540 (18 kDa) and is necessary for the binding of the chaperone with substrate proteins and peptides. The high-resolution NMR solution structure is based on 4150 experimental distance constraints leading to an average root-mean-square precision of 0.38 A for the backbone atoms and 0.76 A for all atoms in the beta-sandwich sub-domain. The protein is observed to bind residue Leu539 in its hydrophobic substrate-binding groove by intramolecular interaction. The position of a helical latch differs dramatically from what is observed in the crystal and solution structures of the homologous prokaryotic chaperone DnaK. In the Hsc70 structure, the helix lies in a hydrophobic groove and is anchored by a buried salt-bridge. Residues involved in this salt-bridge appear to be important for the allosteric functioning of the protein. A mechanism for interdomain allosteric modulation of substrate-binding is proposed. It involves large-scale movements of the helical domain, redefining the location of the hinge area that enables such motions. PMID- 10373375 TI - Structure-activity relationships of omega-conotoxins MVIIA, MVIIC and 14 loop splice hybrids at N and P/Q-type calcium channels. AB - The omega-conotoxins are a set of structurally related, four-loop, six cysteine containing peptides, that have a range of selectivities for different subtypes of the voltage-sensitive calcium channel (VSCC). To investigate the basis of the selectivity displayed by these peptides, we have studied the binding affinities of two naturally occurring omega-conotoxins, MVIIA and MVIIC and a series of 14 MVIIA/MVIIC loop hybrids using radioligand binding assays for N and P/Q-type Ca2+channels in rat brain tissue. A selectivity profile was developed from the ratio of relative potencies at N-type VSCCs (using [125I]GVIA radioligand binding assays) and P/Q-type VSCCs (using [125I]MVIIC radioligand binding assays). In these peptides, loops 2 and 4 make the greatest contribution to VSCC subtype selectivity, while the effects of loops 1 and 3 are negligible. Peptides with homogenous combinations of loop 2 and 4 display clear selectivity preferences, while those with heterogeneous combinations of loops 2 and 4 are less discriminatory. 1H NMR spectroscopy revealed that the global folds of MVIIA, MVIIC and the 14 loop hybrid peptides were similar; however, several differences in local structure were identified. Based on the binding data and the 3D structures of MVIIA, GVIA and MVIIC, we have developed a preliminary pharmacophore based on the omega-conotoxin residues most likely to interact with the N-type VSCC. PMID- 10373376 TI - Identification of the binding interfaces on CheY for two of its targets, the phosphatase CheZ and the flagellar switch protein fliM. AB - CheY is the response regulator protein serving as a phosphorylation-dependent switch in the bacterial chemotaxis signal transduction pathway. CheY has a number of proteins with which it interacts during the course of the signal transduction pathway. In the phosphorylated state, it interacts strongly with the phosphatase CheZ, and also the components of the flagellar motor switch complex, specifically with FliM. Previous work has characterized peptides consisting of small regions of CheZ and FliM which interact specifically with CheY. We have quantitatively measured the binding of these peptides to both unphosphorylated and phosphorylated CheY using fluorescence spectroscopy. There is a significant enhancement of the binding of these peptides to the phosphorylated form of CheY, suggesting that these peptides share much of the binding specificity of the intact targets of the phosphorylated form of CheY. We also have used modern nuclear magnetic resonance methods to characterize the sites of interaction of these peptides on CheY. We have found that the binding sites are overlapping and primarily consist of residues in the C-terminal portion of CheY. Both peptides affect the resonances of residues at the active site, indicating that the peptides may either bind directly at the active site or exert conformational influences that reach to the active site. The binding sites for the CheZ and FliM peptides also overlap with the previously characterized CheA binding interface. These results suggest that interaction with these three proteins of the signal transduction pathway are mutually exclusive. In addition, since these three proteins are sensitive to the phosphorylation state of CheY, it may be that the C terminal region of CheY is most sensitive for the conformational changes occurring upon phosphorylation. PMID- 10373377 TI - Electrostatic contributions to the stability of hyperthermophilic proteins. AB - Electrostatic contributions to the folding free energy of several hyperthermophilic proteins and their mesophilic homologs are calculated. In all the cases studied, electrostatic interactions are more favorable in the hyperthermophilic proteins. The electrostatic free energy is found not to be correlated with the number of ionizable amino acid residues, ion pairs or ion pair networks in a protein, but rather depends on the location of these groups within the protein structure. Moreover, due to the large free energy cost associated with burying charged groups, buried ion pairs are found to be destabilizing unless they undergo favorable interactions with additional polar groups, including other ion pairs. The latter case involves the formation of stabilizing ion pair networks as is observed in a number of proteins. Ion pairs located on the protein surface also provide stabilizing interactions in a number of cases. Taken together, our results suggest that many hyperthermophilic proteins enhance electrostatic interactions through the optimum placement of charged amino acid residues within the protein structure, although different design strategies are used in different cases. Other physical mechanisms are also likely to contribute, however optimizing electrostatic interactions offers a simple means of enhancing stability without disrupting the core residues characteristic of different protein families. PMID- 10373378 TI - Substrate recognition and selectivity of peptide deformylase. Similarities and differences with metzincins and thermolysin. AB - The substrate specificity of Escherichia coli peptide deformylase was investigated by measuring the efficiency of the enzyme to cleave formyl- peptides of the general formula Fo-Xaa-Yaa-NH2, where Xaa represents a set of 27 natural and unusual amino acids and Yaa corresponds to a set of 19 natural amino acids. Substrates with bulky hydrophobic side-chains at the P1' position were the most efficiently cleaved, with catalytic efficiencies greater by two to five orders of magnitude than those associated with polar or charged amino acid side-chains. Among hydrophobic side-chains, linear alkyl groups were preferred at the P1' position, as compared to aryl-alkyl side-chains. Interestingly, in the linear alkyl substituent series, with the exception of norleucine, deformylase exhibits a preference for the substrate containing Met in the P1' position. Next, the influence in catalysis of the second side-chain was studied after synthesis of 20 compounds of the formula Fo-Nle-Yaa-NH2. Their deformylation rates varied within a range of only one order of magnitude. A 3D model of the interaction of PDF with an inhibitor was then constructed and revealed indeed the occurrence of a deep and hydrophobic S1' pocket as well as the absence of a true S2' pocket. These analyses pointed out a set of possible interactions between deformylase and its substrates, which could be the ground driving substrate specificity. The validity of this enzyme:substrate docking was further probed with the help of a set of site-directed variants of the enzyme. From this, the importance of residues at the bottom of the S1' pocket (Ile128 and Leu125) as well as the hydrogen bond network that the main chain of the substrate makes with the enzyme were revealed. Based on the numerous homologies that deformylase displays with thermolysin and metzincins, a mechanism of enzyme:substrate recognition and hydrolysis could finally be proposed. Specific features of PDF with respect to other members of the enzymes with motif HEXXH are discussed. PMID- 10373379 TI - Folding mechanism of Pseudomonas aeruginosa cytochrome c551: role of electrostatic interactions on the hydrophobic collapse and transition state properties. AB - We report on the folding kinetics of the small 82 residue cytochrome c551from Pseudomonas aeruginosa. The presence of two Trp residues (Trp56 and Trp77) allows the monitoring of fluorescence quenching on refolding in two different regions of the protein. A single His residue (the iron-coordinating His16) permits the study of refolding in the absence of miscoordination events. After identification of the kinetic traps (Pro isomerization and aggregation of denatured protein), overall refolding kinetics is described by two processes: (i) a burstphase collapse (faster than milliseconds) which we show to be a global event leading to a state whose compactness depends on the overall net charge; at the isoeletric pH (4.7), it is maximally compact, while above and below it is more expanded; and (ii) an exponential phase (in the millisecond time range) leading to the native protein via a transition state(s) possibly involving the formation of a specific salt bridge between Lys10 and Glu70, at the contact between the N and C-terminal helices. Comparison with the widely studied horse cytochrome c allows the discussion of similarities and differences in the folding of two proteins which have the same "fold" despite a very low degree of sequence homology (<30 %). PMID- 10373381 TI - New Drugs for Asthma IV, 4th International Conference in Konstanz, Germany, 23-25 July, 1998. Proceedings. PMID- 10373380 TI - New efficient statistical sequence-dependent structure prediction of short to medium-sized protein loops based on an exhaustive loop classification. AB - A bank of 13,563 loops from three to eight amino acid residues long, representing motifs between two consecutive regular secondary structures, has been derived from protein structures presenting less than 95 % sequence identity. Statistical analyses of occurrences of conformations and residues revealed length-dependent over-representations of particular amino acids (glycine, proline, asparagine, serine, and aspartate) and conformations (alphaL, epsilon, betaPregions of the Ramachandran plot). A position-dependent distribution of these occurrences was observed for N and C-terminal residues, which are correlated to the nature of the flanking regions. Loops of the same length were clustered into statistically meaningful families on the basis of their backbone structures when placed in a common reference frame, independent of the flanks. These clusters present significantly different distributions of sequence, conformations, and endpoint residue Calphadistances. On the basis of the sequence-structure correlation of this clustering, an automatic loop modeling algorithm was developed. Based on the knowledge of its sequence and of its flank backbone structures each query loop is assigned to a family and target loop supports are selected in this family. The support backbones of these target loops are then adjusted on flanking structures by partial exploration of the conformational space. Loop closure is performed by energy minimization for each support and the final model is chosen among connected supports based upon energy criteria. The quality of the prediction is evaluated by the root-mean-square deviation (rmsd) between the final model and the native loops when the whole bank is re-attributed on itself with a Jackknife test. This average rmsd ranges from 1.1 A for three-residue loops to 3.8 A for eight-residue loops. A few poorly predicted loops are inescapable, considering the high level of diversity in loops and the lack of environment data. To overcome such modeling problems, a statistical reliability score was assigned for each prediction. This score is correlated to the quality of the prediction, in terms of rmsd, and thus improves the selection accuracy of the model. The algorithm efficiency was compared to CASP3 target loop predictions. Moreover, when tested on a test loop bank, this algorithm was shown to be robust when the loops are not precisely delimited, therefore proving to be a useful tool in practice for protein modeling. PMID- 10373382 TI - Unmet needs in the pharmacological treatment of asthma. PMID- 10373383 TI - Strategies for novel COPD therapies. PMID- 10373384 TI - Cytokine inhibitor strategies. PMID- 10373385 TI - Potential role of the endothelins in airway remodelling in asthma. PMID- 10373386 TI - IL-5 inhibition as a therapy for allergic disease. PMID- 10373387 TI - Current animal models are not predictive for clinical asthma. PMID- 10373388 TI - Clinical features of bronchial hyperresponsiveness. PMID- 10373389 TI - Airway hyperreactivity: direct smooth muscle approach. PMID- 10373390 TI - KATP channel openers for the treatment of airways hyperreactivity. PMID- 10373391 TI - Role of leukotrienes and leukotriene receptor antagonists in asthma. PMID- 10373392 TI - Adenosine receptor ligands: potential as therapeutic agents in asthma and COPD. PMID- 10373393 TI - Muscarinic receptor antagonists. PMID- 10373394 TI - The ideal steroid. PMID- 10373396 TI - Ariflo (SB 207499), a second generation phosphodiesterase 4 inhibitor for the treatment of asthma and COPD: from concept to clinic. PMID- 10373395 TI - Therapeutic potential of selective PDE inhibitors in asthma. PMID- 10373397 TI - Adhesion molecule strategies. PMID- 10373398 TI - Strategies for target identification. PMID- 10373399 TI - Immunochemical analysis of liver microsomal cytochromes P450 of the American alligator, Alligator mississippiensis. AB - Ten antibodies raised against various mammalian and fish cytochromes P450 (CYP) enzymes were used to probe the effects of xenobiotic pretreatment on liver microsomes of the American alligator, Alligator mississippiensis. Pretreatment with phenobarbital (PB), 3-methylcholanthrene (3MC), and PB plus 3MC elicited significant induction of multiple CYP enzymes in alligator, as detected by antibodies to CYP1A, CYP2B, CYP2C, CYP2E, CYP2K, and CYP3A. In contrast to the rat, 3MC treatment induced alligator liver microsomes that were immunoreactive with antibodies to CYP2 family enzymes. Induction of CYP enzymes was not as apparent with the Aroclor 1254 (ARO), and 2,2',4,4' tetrachlorobiphenyl (TCB) pretreatment used; fewer CYP enzymes primarily detected with antibodies against CYP2C or CYP2E were observed. Clofibrate (CLO; 80 mg/kg Days 1-4), markedly induced CYP4A in rat but this induction was not apparent in alligator. A purified PB-induced alligator liver microsomal CYP enzyme cross-reacted with several antibodies raised against CYP2 family enzymes but did not cross-react with antibodies raised against other CYP families. This indicates the PB-inducible CYP in alligator shares some epitope homology with several CYP2-family enzymes from other animals. These experiments demonstrate the usefulness and limitations of using antibodies across phylogenetic classes. While indicating the presence of CYP enzymes that have epitope homology with CYP1A, CYP2, CYP3 and CYP4 enzymes in alligator, it remains to be established whether these CYP forms are alligator orthologues of mammalian enzymes. In all cases, the relative abundance of alligator liver microsomal CYP as determined by immunoblot analysis appeared lower than found in rat. The presence and induction of CYP indicated by immunochemical analysis, corroborated previously reported enzymatic studies of the same microsomal preparations (Ertl et al., 1998a). Thus, increases in CYP protein by the various inducers employed were paralled by the increases in CYP enzyme-specific or selective activities, e.g., induction of CYP1A protein corresponded with induction of EROD. PMID- 10373401 TI - Paraoxonase 1 mutations in a Turkish population. AB - Human serum paraoxonase 1 (PON1) catalyzes the hydrolysis of certain organophosphate pesticides and nerve gases and so may alter significantly an individual's susceptibility to the toxicity of these chemicals. Moreover, PON1 hydrolyzes lipid peroxides complexed to low density lipoproteins (LDL) and therefore it was suggested that PON1 may be one of the genes that is involved in the pathogenesis of cardiovascular diseases. Its activity shows interindividual and interethnic variability. At least two mutation sites, namely Gln192Arg (Q/R) and Leu55Met (L/M) were reported responsible for the variations in enzyme activity. The aim of the present study was to determine the frequency of these mutations in Turks and compare the results with other European and Oriental populations. A total of 381 unrelated Turkish individuals were genotyped for Gln192Arg and Leu55Met polymorphisms by PCR-RFLP using AlwI and NlaIII, respectively. Genotype distribution was QQ = 0.49, QR = 0.40, RR = 0.11, and LL = 0.52, LM = 0.39, MM = 0.09. Thus frequencies of high activity alleles R (Arg) and L (Leu) were found as 0.31 and 0.72, respectively. The frequency of these alleles was slightly higher in Turkish subjects than other Caucasian populations but much lower compared to Oriental populations. PMID- 10373400 TI - Inhibition of hCG-stimulated steroidogenesis in cultured mouse Leydig tumor cells by bisphenol A and octylphenols. AB - Some environmental chemicals exhibit estrogenic or antiandrogenic activity. Some of these, such as bisphenol A (bis A) and octylphenols, are used in large amounts in many applications. We have analyzed the effects of bis A and octylphenols on steroidogenesis in Leydig cells by measuring the LH receptor-mediated cAMP and progesterone (P) production in cultured mouse Leydig tumor cells (mLTC-1 cells). After preincubation of mLTC-1 cells for 48 h in the presence of bis A or one of the octylphenols in micromolar concentration, the hCG-stimulated cAMP and P formation in these cells was inhibited. Bis A or octylphenols could neither inhibit cAMP nor P formation stimulated by forskolin (Fk) or cholera toxin (CT) nor steroidogenesis stimulated by 8-Br-cAMP. The preincubation of mLTC-1 cells with estradiol or diethylstilbesterol (DES) at the concentration of 10(-8) mol/liter had no inhibitory effect on cAMP formation stimulated by hCG or Fk but P production was inhibited. Similarly, both estrogens inhibited P production stimulated by 8-Br-cAMP. Bis A or octylphenols had no effect on 125I-hCG binding to Leydig cell LH-receptors. Thus, these environmental chemicals appear to inhibit cAMP formation and steroidogenesis in mLTC-1 Leydig tumor cells by preventing the coupling between LH receptor and the adenylate cyclase. Since, estradiol did not inhibit hCG-stimulated cAMP production, the effects of bis A and octylphenols may not be estrogen related. This emphasizes the complexity of endocrine disruption: chemicals show multiple endocrine activities that may disturb several organs in distinct ways. PMID- 10373402 TI - Effects of hard metal on nitric oxide pathways and airway reactivity to methacholine in rat lungs. AB - To examine whether the development of hard metal (HM)-induced occupational asthma and interstitial lung disease involves alterations in nitric oxide (NO) pathways, we examined the effects of an industrial HM mixture on NO production, interactions between HM and lipopolysaccharide (LPS) on NO pathways, and alterations in airway reactivity to methacholine in rat lungs. HM (2.5 to 5 mg/100 g intratracheal) increased NO synthase (NOS; EC 1.14.23) activity of rat lungs at 24 h without increasing inducible NOS (iNOS) or endothelial NOS (eNOS) mRNA abundance or iNOS, eNOS, or brain NOS (bNOS) proteins. The increase in NOS activity correlated with the appearance histologically of nitrotyrosine immunofluorescence in polymorphonuclear leukocytes (PMN) and macrophages. Intraperitoneal injection of LPS (1 mg/kg) caused up-regulation of iNOS activity, mRNA, and protein at 8 h but not at 24 h. HM at 2.5 mg/100 g, but not at 5 mg/100 g, potentiated the LPS-induced increase in NOS activity, iNOS mRNA, and protein. However, HM decreased eNOS activity at 8 h and eNOS protein at 24 h. Whole body plethysmography on conscious animals revealed that HM caused basal airway obstruction and a marked hyporeactivity to inhaled methacholine by 6-8 h, which intensified over 30-32 h. HM-treatment caused protein leakage into the alveolar space, and edema, fibrin formation, and an increase in the number of inflammatory cells in the lungs and in the bronchoalveolar lavage. These results suggest that a HM-induced increase in NO production by pulmonary inflammatory cells is associated with pulmonary airflow abnormalities in rat lungs. PMID- 10373403 TI - Toxic effects of octylphenol on cultured rat spermatogenic cells and Sertoli cells. AB - Alkylphenols, including the estrogenic 4-tert-octylphenol (OP), are environmental pollutants. Because administration of OP to adult male rats impairs spermatogenesis and OP has been shown to be toxic to aquatic animals and to mammalian splenocytes in vitro, we studied whether OP exerts direct toxic effects on cultured spermatogenic cells and Sertoli cells isolated from male rats. Cell viability was assessed with a Live/Dead Eukolight viability/cytotoxicity kit. Culture of mixed spermatogenic cells from adult rats with 10(-8) M OP or Sertoli cells from 19- to 21-day-old rats with 10(-12) M OP, but not with 0.08% EtOH (vehicle) or 10(-6) M 17beta-estradiol (E2) or dexamethasone (DEX), significantly decreased the percentage of viable cells after 24 h of treatment. None of the treatments significantly altered total cell number. Flow cytometric analyses of spermatogenic cells revealed that exposure to 10(-4) or 10(-6) M OP yielded abnormal relative ploidy classes. Four hours of treatment with 10(-6) M OP, but not with 10(-6) M E2 or DEX, caused significant chromatin condensation in Sertoli cells as observed with acridine orange staining. The decreased percentage of viable cells after 24 h of exposure to 10(-6) M OP remained when Sertoli cells were cultured in Ca2+-free medium. Sertoli cells contained nuclei of reduced size and labeled 3'-OH DNA ends as detected by microscopic analyses when the cells had been incubated for 24 h with 10(-6) M OP but not with vehicle or 10(-6) M E2 or DEX. The results demonstrate that OP, but not E2 or DEX, is directly toxic to cultured rat spermatogenic cells and Sertoli cells and suggest that this toxic effect in Sertoli cells is exerted through Ca2+-independent apoptosis. PMID- 10373404 TI - Induction of the cell cycle regulatory gene p21 (Waf1, Cip1) following methylmercury exposure in vitro and in vivo. AB - Methylmercury (MeHg) is recognized as a significant environmental hazard, particularly to the development of the nervous system. To study the molecular mechanisms underlying cell cycle inhibition by MeHg, we assessed the involvement of p21 (Waf1, Cip1), a cell cycle regulatory gene implicated in the G1 and G2 phases of cell cycle arrest, in primary embryonic cells and adult mice following MeHg exposure. Previous literature has supported the association of increased p21 expression with chondrocyte differentiation. In support of this finding, we observed an increasing p21 expression during limb bud (LB), but not midbrain central nervous system (CNS) cell differentiation. Both embryonic LB and CNS cells responded to MeHg exposure with a concentration-dependent increase in p21 mRNA. In the parallel adult study, C57BL/6 female mice were chronically exposed to 10 ppm MeHg via drinking water for 4 weeks. While there was limited or absent induction of Gadd45, Gadd153, and the gamma-glutamylcysteine synthetase catalytic subunit, p21 was markedly induced in the brain, kidney, and liver tissues in most of the animals that showed MeHg-induced behavioral toxicity such as hyperactivity and tremor. Furthermore, the induction of p21 mRNA was accompanied by an increase in p21 protein level. The results indicate that the activation of cell cycle regulatory genes may be one mechanism by which MeHg interferes with the cell cycle in adult and developing organisms. Continued examination of the molecular mechanisms underlying cell cycle inhibition may potentially lead to utilization of this mechanistic information to characterize the effects of MeHg exposure in vivo. PMID- 10373405 TI - Involvement of nuclear factor-kappaB in a murine model for the acute form of autoimmune-like toxic oil syndrome. AB - The toxic oil syndrome (TOS) represents an exogenously induced autoimmune disease with acute or chronic symptoms similar to systemic lupus erythematosus or scleroderma. When genetically different mouse strains were exposed to oleic acid anilide (OAA), it was possible to mimic the different syndrome manifestations. The aim of the present study was to examine the role of NF-kappaB/Rel transcription factors in the development of the severe acute wasting disease observed in A/J mice. Within a week of OAA exposure, the A/J, but not B10.S strain, displayed weight loss, cachexia, apathy, reduced activity, and breathing difficulties. In affected A/J mice we observed a marked increase in NF-kappaB activation (p50/p65 dimers) both in splenic T cells and peritoneal macrophages as well as in tissue from aorta and gut. Incubation of splenocytes with OAA in vitro induced a dose-dependent removal of IkappaB-alpha, accompanied by NF-kappaB activation, whereas Sp-1 binding was not affected. Furthermore, we demonstrated the increased expression of the two NF-kappaB target genes IL-6 and IL-1beta in OAA-exposed mice and a transient OAA-induced accumulation of TNFalpha in vitro. This is the first report which implicates NF-kappaB/Rel in acute forms of chemically induced autoimmune-like disease and may serve as a paradigm for the involvement of this transcriptional system in acute processes associated with autoimmunity, suggesting possible avenues of therapeutic intervention. PMID- 10373406 TI - Neurotoxic potentiation is related to a metabolic interaction between p bromophenylacetylurea and phenylmethanesulfonyl fluoride. AB - This study investigated the neurotoxic potentiation and metabolic interaction between p-bromophenylacetylurea (BPAU) and phenylmethanesulfonyl fluoride (PMSF). The results showed that F344 rats given two successive daily doses of 150 mg/kg BPAU developed a moderate degree of ataxia. When rats were coadministrated a single intraperitoneal dose of 100 mg/kg PMSF either 1 day before, or 4 h or 1 day after the two daily doses of BPAU, the severity of ataxia was significantly increased. No such effect was observed when PMSF was given 4 days after BPAU, although this time point was still prior to the development of the neuropathy. The enhancement or potentiation of neuropathy by PMSF was thus seen only at times when parent BPAU was present in the target tissues. A pharmacokinetic study showed that PMSF increased the concentrations of BPAU and its metabolite, N' hydroxy-p-bromophenylacetylurea (M1), in tissues and decreased the concentration of the metabolite 4-(4-bromophenyl)-3-oxapyrrolidine-2,5-dione (M2) in serum. This indicated that PMSF inhibited the M2 pathway and more BPAU was metabolized via the M1 pathway. This increased both BPAU and M1 levels in tissues and hence would have increased BPAU-induced neurotoxicity. We conclude that PMSF does not need to act directly on target sites to potentiate BPAU-induced neurotoxicity, since its interference with BPAU metabolism was sufficient to account for the increase in BPAU neurotoxicity. Thus a metabolic interaction underlies the neurotoxic potentiation between these two compounds rather than the target site interaction seen between PMSF and neuropathic organophosphates. This study is the first to demonstrate that interference with the metabolism of BPAU is an important aspect of the potentiation of BPAU-induced neurotoxicity. PMID- 10373408 TI - Chemical index for volume 157 PMID- 10373407 TI - Association of low PON1 type Q (type A) arylesterase activity with neurologic symptom complexes in Gulf War veterans. AB - Previously Haley et al. described six possible syndromes identified by factor analysis of symptoms in Gulf War veterans and demonstrated that veterans with these symptom complexes were more neurologically impaired than age-sex-education matched well controls. They also uncovered strong associations (relative risks 4 8) suggesting that these symptom complexes were related to wartime exposure to combinations of organophosphate pesticides, chemical nerve agents, high concentration DEET insect repellant, and symptoms of advanced acute toxicity after taking pyridostigmine. Here we have shown that compared to controls, ill veterans with the neurologic symptom complexes were more likely to have the R allele (heterozygous QR or homozygous R) than to be homozygous Q for the paraoxonase/arylesterase 1 (PON1) gene. Moreover, low activity of the PON1 type Q (Gln192, formerly designated type A) arylesterase allozyme distinguished ill veterans from controls better than just the PON1 genotype or the activity levels of the type R (Arg192, formerly designated type B) arylesterase allozyme, total arylesterase, total paraoxonase, or butyrylcholinesterase. A history of advanced acute toxicity after taking pyridostigmine was also correlated with low PON1 type Q arylesterase activity. Type Q is the allozyme of paraoxonase/arylesterase that most efficiently hydrolyzes several organophosphates including sarin, soman, and diazinon. These findings further support the proposal that neurologic symptoms in some Gulf War veterans were caused by environmental chemical exposures. PMID- 10373409 TI - Signaling pathways activated by oncogenic forms of Abl tyrosine kinase. PMID- 10373410 TI - The head domain of plakophilin-1 binds to desmoplakin and enhances its recruitment to desmosomes. Implications for cutaneous disease. AB - The contribution of desmosomes to epidermal integrity is evident in the inherited blistering disorder associated with the absence of a functional gene for plakophilin-1. To define the function of plakophilin-1 in desmosome assembly, interactions among the desmosomal cadherins, desmoplakin, and the armadillo family members plakoglobin and plakophilin-1 were examined. In transient expression assays, plakophilin-1 formed complexes with a desmoplakin amino terminal domain and enhanced its recruitment to cell-cell borders; this recruitment was not dependent on the equimolar expression of desmosomal cadherins. In contrast to desmoplakin-plakoglobin interactions, the interaction between desmoplakin and plakophilin-1 was not mediated by the armadillo repeat domain of plakophilin-1 but by the non-armadillo head domain, as assessed by yeast two-hybrid and recruitment assays. We propose a model whereby plakoglobin serves as a linker between the cadherins and desmoplakin, whereas plakophilin-1 enhances lateral interactions between desmoplakin molecules. This model suggests that epidermal lesions in patients lacking plakophilin-1 are a consequence of the loss of integrity resulting from a decrease in binding sites for desmoplakin and intermediate filaments at desmosomes. PMID- 10373411 TI - Homodimerization of soluble guanylyl cyclase subunits. Dimerization analysis using a glutathione s-transferase affinity tag. AB - Soluble guanylyl cyclase (sGC) is an alpha/beta-heterodimeric hemoprotein that, upon interaction with the intercellular messenger molecule NO, generates cGMP. Although the related family of particulate guanylyl cyclases (pGCs) forms active homodimeric complexes, it is not known whether homodimerization of sGC subunits occurs. We report here the expression in Sf9 cells of glutathione S-transferase tagged recombinant human sGCalpha1 and beta1 subunits, applying a novel and rapid purification method based on GSH-Sepharose affinity chromatography. Surprisingly, in intact Sf9 cells, both homodimeric GSTalpha/alpha and GSTbeta/beta complexes were formed that were catalytically inactive. Upon coexpression of the respective complementary subunits, GSTalpha/beta or GSTbeta/alpha heterodimers were preferentially formed, whereas homodimers were still detectable. When subunits were mixed after expression, e.g. GSTbeta and beta or GSTalpha and beta, no dimerization was observed. In conclusion, our data suggest the previously unrecognized possibility of a physiological equilibrium between homo- and heterodimeric sGC complexes. PMID- 10373412 TI - Agonist-dependent interaction of the rat somatostatin receptor subtype 2 with cortactin-binding protein 1. AB - We report here an interaction between the C terminus of the rat somatostatin receptor subtype 2 (SSTR2) and a protein that has recently been identified as cortactin-binding protein 1 (CortBP1). Interaction is mediated by the PDZ (PSD 95/discs large/ZO-1) domain of CortBP1. As shown by in situ hybridization, SSTR2 and cortactin-binding protein are coexpressed in the rat brain. The association between SSTR2 and the PDZ-domain of CortBP1 was verified by overlay assays and by coprecipitation after transfection in human embryonic kidney (HEK) cells. Analysis by confocal microscopy indicates that CortBP1 is distributed diffusely throughout the cytosol in transfected cells and that it becomes concentrated at the plasma membrane when SSTR2 is present. This process is largely increased when the receptor is stimulated by somatostatin; as CortBP1 interacts with the C terminus of SSTR2, our data suggest that the binding of agonist to the receptor increase the accessibility of the receptor C terminus to the PDZ domain of CortBP1. Our data for the first time establish a link between a G-protein coupled receptor and constituents of the cytoskeleton. PMID- 10373413 TI - Catalytic mechanism and function of invariant glutamic acid 173 from the histone acetyltransferase GCN5 transcriptional coactivator. AB - Within chromatin, reversible acetylation of core histones is critical for transcriptional activation of eukaryotic target genes. The recent identification of intrinsic histone acetyltransferase (HAT) catalytic activity from a number of transcriptional co-activators (including yeast GCN5, p300/CBP, P/CAF, and TAFII250), has underscored the importance of protein acetylation in transcriptional control. The GCN5 family is the prototype for a diverse group of at least four distinct human HATs families. Although there is now a clear link between in vivo HAT catalytic activity and gene activation, little is known about the molecular mechanisms of histone acetylation. Herein, we report the first detailed biochemical study that probes the catalytic mechanism and the function of invariant glutamic acid 173 within the GCN5 family of HATs. Our results suggest that the HAT reaction involves the formation of a ternary complex (histones, acetyl-CoA, and enzyme) where the epsilon-amino group of histone lysine residues directly attacks the bound acetyl-CoA. The acetylation reaction requires deprotonation of the epsilon-amino group prior to nucleophilic attack. Employing site-directed mutagenesis, chemical modification, steady-state, and pH dependent rate analysis, it is demonstrated that glutamic acid 173 is an essential catalytic residue, acting as a general base catalyst by deprotonating the histone substrate. PMID- 10373414 TI - Apocalmodulin binds to the myosin light chain kinase calmodulin target site. AB - The interaction of a 20-residue-long peptide derived from the calmodulin-binding domain of the smooth muscle myosin light chain kinase with calcium-free calmodulin (apocalmodulin) was studied using a combination of isothermal titration calorimetry and differential scanning calorimetry. We showed that: (i) a significant binding between apocalmodulin and the target peptide (RS20) exists in the absence of salt (Ka = 10(6) M-1), (ii) the peptide interacts with the C terminal lobe of calmodulin and adopts a partly helical conformation, and (iii) the presence of salt weakens the affinity of the peptide for apocalmodulin, emphasizing the importance of electrostatic interactions in the complex. Based on these results and taking into account the work of Bayley et al. (Bayley, P. M., Findlay, W.A., and Martin, S. R. (1996) Protein Sci. 5, 1215-1228), we suggest a physiological role for apocalmodulin. PMID- 10373415 TI - High density O-glycosylation on tandem repeat peptide from secretory MUC1 of T47D breast cancer cells. AB - The site-specific O-glycosylation of MUC1 tandem repeat peptides from secretory mucin of T47D breast cancer cells was analyzed. After affinity isolation on immobilized BC3 antibody, MUC1 was partially deglycosylated by enzymatic treatment with alpha-sialidase/beta-galactosidase and fragmented by proteolytic cleavage with the Arg-C-specific endopeptidase clostripain. The PAP20 glycopeptides were isolated by reversed phase high pressure liquid chromatography and subjected to the structural analyses by quadrupole time-of-flight electrospray ionization mass spectrometry and to the sequencing by Edman degradation. All five positions of the repeat peptide were revealed as O glycosylation targets in the tumor cell, including the Thr within the DTR motif. The degree of substitution was estimated to average 4.8 glycans per repeat, which compares to 2.6 glycosylated sites per repeat for the mucin from milk (Muller, S., Goletz, S., Packer, N., Gooley, A. A., Lawson, A. M., and Hanisch, F.-G. (1997) J. Biol. Chem. 272, 24780-24793). In addition to a modification by glycosylation, the immunodominant DTR motif on T47D-MUC1 is altered by amino acid replacements (PAPGSTAPAAHGVTSAPESR), which were revealed in about 50% of PAP20 peptides. The high incidence of these replacements and their detection also in other cancer cell lines imply that the conserved tandem repeat domain of MUC1 is polymorphic with respect to the peptide sequence. PMID- 10373416 TI - CAST, a novel CD3epsilon-binding protein transducing activation signal for interleukin-2 production in T cells. AB - Antigen recognition through T cell receptor (TCR)-CD3 complex transduces signals into T cells, which regulate activation, function, and differentiation of T cells. The TCR-CD3 complex is composed of two signaling modules represented by CD3zeta and CD3epsilon. Signaling through CD3zeta has been extensively analyzed, but that via CD3epsilon, which is also crucial in immature thymocyte development, is still not clearly understood. We isolated cDNA encoding a novel CD3epsilon binding protein CAST. CAST specifically interacts in vivo and in vitro with CD3epsilon but not with CD3zeta or FcRgamma via a unique membrane-proximal region of CD3epsilon. CAST is composed of 512 amino acids including a single tyrosine and undergoes tyrosine phosphorylation upon TCR stimulation. Overexpression of two dominant-negative types of CAST, a minimum CD3epsilon-binding domain and a tyrosine-mutant, strongly suppressed NFAT activation and interleukin-2 production. These results demonstrate that CAST serves as a component of preformed TCR complex and transduces activation signals upon TCR stimulation and represents a new signaling pathway via the CD3epsilon-containing TCR signaling module. PMID- 10373417 TI - In vitro oligomerization of a membrane protein complex. liposome-based reconstitution of trimeric photosystem I from isolated monomers. AB - Many membrane proteins can be isolated in different oligomeric forms. Photosystem I (PSI), for example, exists in cyanobacteria either as a monomeric or as a trimeric complex. Neither the factors responsible for the specific trimerization process nor its biological role are known at present. In the filamentous cyanobacterium Spirulina platensis, trimers in contrast to monomers show chlorophyll fluorescence emission at 760 nm. To investigate the oligomerization process as well as the nature of the long wavelength chlorophylls, we describe here an in vitro reconstitution procedure to assemble trimeric PS I from isolated purified PS I monomers. Monomers (and trimers) were extracted from S. platensis with n-dodecyl beta-D-maltoside and further purified by perfusion chromatography steps. The isolated complexes had the same polypeptide composition as other cyanobacteria (PsaA-PsaF and PsaI-PsaM), as determined from high resolution gels and immunoblotting. They were incorporated into proteoliposomes, which had been prepared by the detergent absorption method, starting from a phosphatidylcholine:phosphatidic acid mixture solubilized by octylglucoside. After the addition of monomeric PS I (lipid:chlorophyll, 25:1), octylglucoside was gradually removed by the stepwise addition of Biobeads. The 77 K fluorescence emission spectrum of these proteoliposomes displays a long wavelength emission at 760 nm that is characteristic of PS I trimers, which indicates for the first time the successful in vitro reconstitution of PS I trimers. In addition, a high performance liquid chromatography analysis of complexes extracted from these proteoliposomes confirms the formation of structural trimers. We also could show with this system 1) that at least one of the stromal subunits PsaC, -D, and -E is necessary for trimer formation and 2) that the extreme long wavelength emitting chlorophyll is formed as a result of trimer formation. PMID- 10373418 TI - Mapping subdomains in the C-terminal region of troponin I involved in its binding to troponin C and to thin filament. AB - Troponin I (TnI) is the inhibitory component of troponin, the ternary complex that regulates skeletal and cardiac muscle contraction. Previous work showed that the C-terminal region of TnI, when linked to the "inhibitory region" (residues 98 116), possesses the major regulatory functions of the molecule (Farah, C. S., Miyamoto, C. A., Ramos, C. H. I., Silva, A. C. R., Quaggio, R. B., Fujimori, K., Smillie, L. B., and Reinach, F. C. (1994) J. Biol. Chem. 269, 5230-5240). To investigate these functions in more detail, serial deletion mutants of the C terminal region of TnI were constructed. These experiments showed that longer C terminal deletions result in lower inhibition of the actomyosin ATPase activity and weaken the interaction with the N-terminal domain of troponin C (TnC), consistent with the antiparallel model for the interaction between these two proteins. The conclusion is that the whole C-terminal region of TnI is necessary for its full regulatory activity. The region between residues 137 and 144, which was shown to have homology with residues 108-115 in the inhibitory region (Farah, C. S., and Reinach, F. C. (1995) FASEB J. 9, 755-767), is involved in the binding to TnC. The region between residues 98 and 129 is involved in modulating the affinity of TnC for calcium. The C-terminal residues 166-182 are involved in the binding of TnI to thin filament. A model for the function of TnI is discussed. PMID- 10373419 TI - Ferric alpha-hydroxyheme bound to heme oxygenase can be converted to verdoheme by dioxygen in the absence of added reducing equivalents. AB - Whether or not reducing equivalents are indispensable for the conversion of ferric alpha-hydroxyheme bound to heme oxygenase-1 to verdoheme remains controversial (Matera, K. M., Takahashi, S., Fujii, H., Zhou, H., Ishikawa, K., Yoshimura, T., Rousseau, D. L., Yoshida, T., and Ikeda-Saito, M. (1996) J. Biol. Chem. 271, 6618-6624; Liu, Y., Moenne-Loccoz, P., Loehr, T. M., and Ortiz de Montellano, P. R. (1997) J. Biol. Chem. 272, 6906-6917). To resolve this controversy, we have prepared a ferric alpha-hydroxyheme-heme oxygenase-1 complex and titrated the complex with O2 under strictly anaerobic conditions. The formation of verdoheme was monitored by optical and electron spin resonance spectroscopies. Electron spin resonance spectra of the complex showed that alpha hydroxyheme exists as a mixture of resonance structures composed of the iron(III) porphyrin and the iron(II) porphyrin pi neutral radical. Upon addition of CO the latter species becomes dominant. The results obtained from these titration experiments indicate that alpha-hydroxyheme can be converted to verdoheme by an approximately equimolar amount of O2 without any requirement for exogenous electrons. The verdoheme formed from alpha-hydroxyheme was shown to be in the ferrous oxidation state by the addition of CO or potassium ferricyanide to the resultant verdoheme-heme oxygenase-1 complex. PMID- 10373420 TI - The oxidized forms of dATP are substrates for the human MutT homologue, the hMTH1 protein. AB - The possibility that Escherichia coli MutT and human MTH1 (hMTH1) hydrolyze oxidized DNA precursors other than 8-hydroxy-dGTP (8-OH-dGTP) was investigated. We report here that hMTH1 hydrolyzed 2-hydroxy-dATP (2-OH-dATP) and 8-hydroxy dATP (8-OH-dATP), oxidized forms of dATP, but not (R)-8,5'-cyclo-dATP, 5-hydroxy dCTP, and 5-formyl-dUTP. The kinetic parameters indicated that 2-OH-dATP was hydrolyzed more efficiently and with higher affinity than 8-OH-dGTP. 8-OH-dATP was hydrolyzed as efficiently as 8-OH-dGTP. The preferential hydrolysis of 2-OH dATP over 8-OH-dGTP was observed at all of the pH values tested (pH 7.2 to pH 8.8). In particular, a 5-fold difference in the hydrolysis efficiencies for 2-OH dATP over 8-OH-dGTP was found at pH 7.2. However, E. coli MutT had no hydrolysis activity for either 2-OH-dATP or 8-OH-dATP. Thus, E. coli MutT is an imperfect counterpart for hMTH1. Furthermore, we found that 2-hydroxy-dADP and 8-hydroxy dGDP competitively inhibited both the 2-OH-dATP hydrolase and 8-OH-dGTP hydrolase activities of hMTH1. The inhibitory effects of 2-hydroxy-dADP were 3-fold stronger than those of 8-hydroxy-dGDP. These results suggest that the three damaged nucleotides share the same recognition site of hMTH1 and that it is a more important sanitization enzyme than expected thus far. PMID- 10373421 TI - On the biosynthesis of alternating alpha-2,9/alpha-2,8 heteropolymer of sialic acid catalyzed by the sialyltransferase of Escherichia coli Bos-12. AB - Escherichia coli Bos-12 synthesizes a heteropolymer of sialic acids with alternating alpha-2,9/alpha-2,8 glycosidic linkages (1). In this study, we have shown that the polysialyltransferase of the E. coli Bos-12 recognizes an alpha 2,8 glycosidic linkage of sialic acid at the nonreducing end of an exogenous acceptor of either the alpha-2,8 homopolymer of sialic acid or the alternating alpha-2,9/alpha-2,8 heteropolymer of sialic acid and catalyzes the transfer of Neu5Ac from CMP-Neu5Ac to this residue. When the exogenous acceptor is an alpha 2,8-linked oligomer of sialic acid, the main product synthesized is derived from the addition of a single residue of [14C]Neu5Ac to form either an alpha-2,8 glycosidic linkage or an alpha-2,9 glycosidic linkage at the nonreducing end, at an alpha-2, 8/alpha-2,9 ratio of approximately 2:1. When the acceptor is the alternating alpha-2,9/alpha-2,8 heteropolymer of sialic acid, chain elongation takes place four to five times more efficiently than the alpha-2,8-linked homopolymer of sialic acid as an acceptor. It was found that the alpha-2,9-linked homopolymer of sialic acid and the alpha-2,8/alpha-2,9-linked hetero-oligomer of sialic acid with alpha-2,9 at the nonreducing end not only failed to serve as an acceptor for the E. coli Bos-12 polysialyltransferase for the transfer of [14C]Neu5Ac, but they inhibited the de novo synthesis of polysialic acid catalyzed by this enzyme. The results obtained in this study favor the proposal that the biosynthesis of the alpha-2, 9/alpha-2,8 heteropolymer of sialic acid catalyzed by the E. coli Bos-12 polysialyltransferase involves a successive transfer of a preformed alpha-2,8-linked dimer of sialic acid at the nonreducing terminus of the acceptor to form an alpha-2,9 glycosidic linkage between the incoming dimer and the acceptor. The glycosidic linkage at the nonreducing end of the alternating alpha-2,9/alpha-2,8 heteropolymer of sialic acid produced by E. coli Bos-12 should be an alpha-2,8 glycosidic bond and not an alpha-2,9 glycosidic linkage. PMID- 10373422 TI - L-selectin interactions with novel mono- and multisulfated Lewisx sequences in comparison with the potent ligand 3'-sulfated Lewisa. AB - The cell adhesion molecule L-selectin binds to 3'-sialyl-Lewis (Le)x and -Lea and to 3'-sulfo-Lex and -Lea sequences. The binding to 3'-sialyl-Lex is strongly affected by the presence of 6-O-sulfate as found on oligosaccharides of the counter receptor, GlyCAM-1; 6-O-sulfate on the N-acetylglucosamine (6-sulfation) enhances, whereas 6-O-sulfate on the galactose (6'-sulfation) virtually abolishes binding. To extend knowledge on the specificity of L-selectin, we have investigated interactions with novel sulfo-oligosaccharides based on the Lex pentasaccharide sequence. We observe that, also with 3'-sulfo-Lex, the 6 sulfation enhances and 6'-sulfation suppresses L-selectin binding. The 6' sulfation without 3'-sialyl or 3'-sulfate gives no binding signal with L selectin. Where the 6-sulfo,3'-sialyl-Lex is on an extended di-N acetyllactosamine backbone, additional 6-O-sulfates on the inner galactose and inner N-acetylglucosamine do not influence the binding. Although binding to the 6,3'-sulfo-Lex and 6-sulfo, 3'-sialyl-Lex sequences is comparable, the former is a more effective inhibitor of L-selectin binding. This difference is most apparent when L-selectin is in paucivalent form (predominantly di- and tetramer) rather than multivalent. Indeed, as inhibitors of the paucivalent L-selectin, the 3'-sulfo-Lex series are more potent than the corresponding 3'-sialyl-Lex series. Thus, for synthetic strategies to design therapeutic oligosaccharide analogs as antagonists of L-selectin binding, those based on the simpler 3'-sulfo-Lex (and also the 3'-sulfo-Lea) would seem most appropriate. PMID- 10373423 TI - A large non-immunized human Fab fragment phage library that permits rapid isolation and kinetic analysis of high affinity antibodies. AB - We report the design, construction, and use of the first very large non-immunized phage antibody library in Fab format, which allows the rapid isolation and affinity analysis of antigen-specific human antibody fragments. Individually cloned heavy and light chain variable region libraries were combined in an efficient two-step cloning procedure, permitting the cloning of a total of 3.7 x 10(10) independent Fab clones. The performance of the library was determined by the successful selection of on average 14 different Fabs against 6 antigens tested. These include tetanus toxoid, the hapten phenyl-oxazolone, the breast cancer-associated MUC1 antigen, and three highly related glycoprotein hormones: human chorionic gonadotropin, human luteinizing hormone, and human follicle stimulating hormone. In the latter category, a panel of either homone-specific or cross-reactive antibodies were identified. The design of the library permits the monitoring of selections with polyclonal phage preparations and to carry out large scale screening of antibody off-rates with unpurified Fab fragments on BIAcore. Antibodies with off-rates in the order of 10(-2) to 10(-4) s-1 and affinities up to 2.7 nM were recovered. The kinetics of these phage antibodies are of the same order of magnitude as antibodies associated with a secondary immune response. This new phage antibody library is set to become a valuable source of antibodies to many different targets, and to play a vital role in target discovery and validation in the area of functional genomics. PMID- 10373424 TI - Molecular cloning of cDNA for matriptase, a matrix-degrading serine protease with trypsin-like activity. AB - A major protease from human breast cancer cells was previously detected by gelatin zymography and proposed to play a role in breast cancer invasion and metastasis. To structurally characterize the enzyme, we isolated a cDNA encoding the protease. Analysis of the cDNA reveals three sequence motifs: a carboxyl terminal region with similarity to the trypsin-like serine proteases, four tandem cysteine-rich repeats homologous to the low density lipoprotein receptor, and two copies of tandem repeats originally found in the complement subcomponents C1r and C1s. By comparison with other serine proteases, the active-site triad was identified as His-484, Asp-539, and Ser-633. The protease contains a characteristic Arg-Val-Val-Gly-Gly motif that may serve as a proteolytic activation site. The bottom of the substrate specificity pocket was identified to be Asp-627 by comparison with other trypsin-like serine proteases. In addition, this protease exhibits trypsin-like activity as defined by cleavage of synthetic substrates with Arg or Lys as the P1 site. Thus, the protease is a mosaic protein with broad spectrum cleavage activity and two potential regulatory modules. Given its ability to degrade extracellular matrix and its trypsin-like activity, the name matriptase is proposed for the protease. PMID- 10373425 TI - Purification and characterization of a complex containing matriptase and a Kunitz type serine protease inhibitor from human milk. AB - Matriptase, a trypsin-like serine protease with two potential regulatory modules (low density lipoprotein receptor and complement C1r/s domains), was initially purified from T-47D breast cancer cells. Given its plasma membrane localization, extracellular matrix-degrading activity, and expression by breast cancer cells, this protease may be involved in multiple aspects of breast tumor progression, including cancer invasion. In breast cancer cells, matriptase was detected mainly as an uncomplexed form; however, low levels of matriptase were detected in complexes. In striking contrast, only the complexed matriptase was detected in human milk. The complexed matriptase has now been purified. Amino acid sequences obtained from the matriptase-associated proteins reveal that they are fragments of a Kunitz-type serine protease inhibitor that was previously reported to be an inhibitor of the hepatocyte growth factor activator. In addition, matriptase and its complexes were detected in milk-derived, SV40 T-antigen-immortalized mammary luminal epithelial cell lines, but not in human foreskin fibroblasts or in HT 1080 fibrosarcoma cells. These results suggest that the milk-derived matriptase complexes are likely to be produced by the epithelial components of the lactating mammary gland in vivo and that the activity and function of matriptase may be differentially regulated by its cognate inhibitor, comparing breast cancer with the lactating mammary gland. PMID- 10373426 TI - Mechanisms involved in the regulation of free fatty acid release from isolated human fat cells by acylation-stimulating protein and insulin. AB - The effects of acylation-stimulating protein (ASP) and insulin on free fatty acid (FFA) release from isolated human fat cells and the signal transduction pathways to induce these effects were studied. ASP and insulin inhibited basal and norepinephrine-induced FFA release by stimulating fractional FFA re esterification (both to the same extent) and by inhibiting FFA produced during lipolysis (ASP to a lesser extent than insulin). Protein kinase C inhibition influenced none of the effects of ASP or insulin. Phosphatidylinositol 3-kinase inhibition counteracted the effects of insulin but not of ASP. Phosphodiesterase 3 (PDE3) activity was stimulated by ASP and insulin, whereas PDE4 activity was slightly increased by ASP only. Selective PDE3 inhibition reversed the effects of both ASP and insulin on fractional FFA re-esterification and lipolysis. Selective PDE4 inhibition slightly counteracted the ASP but not the effect of insulin on fractional FFA re-esterification and did not prevent the action of ASP or insulin on lipolysis. Thus, ASP and insulin play a major role in regulating FFA release from fat cells as follows: insulin by stimulating fractional FFA re esterification and inhibiting lipolysis and ASP mainly by stimulating fractional FFA re-esterification. For both ASP and insulin these effects on FFA release are mediated by PDE3, and for ASP PDE4 might also be involved. The signaling pathway preceding PDE is not known for ASP but involves phosphatidylinositol 3-kinase for insulin. PMID- 10373427 TI - The NAD(P)H:flavin oxidoreductase from Escherichia coli. Evidence for a new mode of binding for reduced pyridine nucleotides. AB - The NAD(P)H:flavin oxidoreductase from Escherichia coli, named Fre, is a monomer of 26.2 kDa that catalyzes the reduction of free flavins using NADPH or NADH as electron donor. The enzyme does not contain any prosthetic group but accommodates both the reduced pyridine nucleotide and the flavin in a ternary complex prior to oxidoreduction. The specificity of the flavin reductase for the pyridine nucleotide was studied by steady-state kinetics using a variety of NADP analogs. Both the nicotinamide ring and the adenosine part of the substrate molecule have been found to be important for binding to the polypeptide chain. However, in the case of NADPH, the 2'-phosphate group destabilized almost completely the interaction with the adenosine moiety. Moreover, NADPH and NMNH are very good substrates for the flavin reductase, and we have shown that both these molecules bind to the enzyme almost exclusively by the nicotinamide ring. This provides evidence that the flavin reductase exhibits a unique mode for recognition of the reduced pyridine nucleotide. In addition, we have shown that the flavin reductase selectively transfers the pro-R hydrogen from the C-4 position of the nicotinamide ring and is therefore classified as an A-side-specific enzyme. PMID- 10373428 TI - Characterization of the mouse sulfonylurea receptor 1 promoter and its regulation. AB - The ATP-sensitive potassium channels (K+ATP channels) are heteromultimeric structures formed by a member of the sulfonylurea receptor (SUR) family and a member of the inwardly rectifying potassium channel family (Kir6.x). The K+ATP channels play an essential role in nutrient-induced insulin secretion from the pancreatic beta-cell. We have cloned and characterized the promoter region of the mouse SUR1 gene, and have shown that it lacks CAAT and TATA boxes or an initiator element. Studies of transcription initiation in several tissues showed that there is a common SUR1 promoter in brain, heart, and pancreas and in the pancreatic beta-cell line, betaTC3. The SUR1 gene uses multiple transcription start sites with the major site located 54 base pairs 5'-upstream of the translation initiation site. Transient transfection experiments in pancreatic beta-cell lines showed that the proximal promoter fragment -84/+54 is sufficient for significant transcriptional activity. The proximal promoter region contains multiple SP1 binding sites, and cotransfection experiments of the SUR1 promoter-luciferase vector with SP1 expression vector in Drosophila SL2 cells demonstrated a stimulatory effect of SP1 on SUR1 transcriptional activity. The mobility shift assays confirmed the interaction of the SP1 transcription factor with the proximal promoter region of the SUR1 gene. Together, these results indicate that SP1 may mediate transcription initiation of the SUR1 gene. In addition, we have described the coordinate regulation of the gene expression of both K+ATP channel subunits by glucocorticoids. SUR1 and Kir6.2 mRNA levels are down-regulated by approximately 40-50% in response to glucocorticoid treatment. Interestingly, the extent of the inhibitory effect as well as the kinetics and sensitivity are very similar for both mRNAs. Studies of mRNA turnover demonstrate that glucocorticoids most likely decrease the transcriptional activity of both SUR1 and Kir6.2 genes since glucocorticoids failed to affect the stability of each mRNA. Likewise, the reduction in mRNA levels was correlated with a decrease in SUR1 and Kir6.2 protein levels. PMID- 10373429 TI - Size and charge requirements for kinetic modulation and actin binding by alkali 1 type myosin essential light chains. AB - The alkali 1-type isoforms of myosin essential light chains from vertebrate striated muscles have an additional 40 or so amino acids at their N terminus compared with the alkali 2-type. Consequently two light chain isoenzymes of myosin subfragment-1 can be isolated. Using synthesized peptide mimics of the N terminal region of alkali 1-type essential light chains, we have found by 1H NMR that the major actin binding region occurred in the N-terminal four residues, APKK. These results were confirmed by mutating this region of the human atrial essential light chain, resulting in altered actin-activated MgATPase kinetics when the recombinant light chains were hybridized into rabbit skeletal subfragment 1. Substitution of either Lys3 or Lys4 with Ala resulted in increased Km and kcat and decreased actin binding (as judged by chemical cross-linking). Replacement of Lys4 with Asp reduced actin binding and increased Km and kcat still further. Alteration of Ala1 to Val did not alter the kinetic parameters of the hybrid subfragment 1 or the essential light chain's ability to bind actin. Furthermore, we found a significant correlation between the apparent Km for actin and the kcat for MgATP turnover for each mutant hybrid, strengthening our belief that the binding of actin by alkali 1-type essential light chains results directly in modulation of the myosin motor. PMID- 10373430 TI - The p56(lck)-interacting protein p62 stimulates transcription via the SV40 enhancer. AB - p62 is a recently identified ubiquitin-binding, cytosolic phosphoprotein that interacts with several signal transduction molecules including the tyrosine kinase p56(lck) and the protein kinase C-zeta. p62 is therefore suggested to serve an important role in signal transduction in the cell, although the physiological function of p62 remains undefined. Here we demonstrate by transient transfection assays that p62 stimulates the transcription of reporter genes linked to the simian virus 40 (SV40) enhancer. A putative p62-responsive element was localized to the B domain of the distal 72-base pair repeat of the SV40 enhancer. p62 was unable to bind this element in vitro, nor was it able to activate transcription when directly tethered to a promoter, suggesting that p62 stimulates transcription via an indirect mechanism. Stimulation of transcription mediated by p62 was dependent on its amino-terminal region, which is also necessary for interaction with cell surface signaling molecules. These findings indicate that p62 may link extracellular signals directly to transcriptional responses, and identify the SV40 enhancer as a downstream target for signal transduction pathways in which p62 participates. PMID- 10373431 TI - Identification of TATA-binding protein-free TAFII-containing complex subunits suggests a role in nucleosome acetylation and signal transduction. AB - Recently we identified a novel human (h) multiprotein complex, called TATA binding protein (TBP)-free TAFII-containing complex (TFTC), which is able to nucleate RNA polymerase II transcription and can mediate transcriptional activation. Here we demonstrate that TFTC, similar to other TBP-free TAFII complexes (yeast SAGA, hSTAGA, and hPCAF) contains the acetyltransferase hGCN5 and is able to acetylate histones in both a free and a nucleosomal context. The recently described TRRAP cofactor for oncogenic transcription factor pathways was also characterized as a TFTC subunit. Furthermore, we identified four other previously uncharacterized subunits of TFTC: hADA3, hTAFII150, hSPT3, and hPAF65beta. Thus, the polypeptide composition of TFTC suggests that TFTC is recruited to chromatin templates by activators to acetylate histones and thus may potentiate initiation and activation of transcription. PMID- 10373432 TI - The subcellular localizations of atypical synaptotagmins III and VI. Synaptotagmin III is enriched in synapses and synaptic plasma membranes but not in synaptic vesicles. AB - Multiple synaptotagmins are expressed in brain, but only synaptotagmins I and II have known functions in fast, synchronous Ca2+-triggered neurotransmitter release. Synaptotagmin III was proposed to regulate other aspects of synaptic vesicle exocytosis, particularly its slow component. Such a function predicts that synaptotagmin III should be an obligatory synaptic vesicle protein, as would also be anticipated from its high homology to synaptotagmins I and II. To test this hypothesis, we studied the distribution, developmental expression, and localization of synaptotagmin III and its closest homolog, synaptotagmin VI. We find that synaptotagmins III and VI are present in all brain regions in heterogeneous distributions and that their levels increase during development in parallel with synaptogenesis. Furthermore, we show by immunocytochemistry that synaptotagmin III is concentrated in synapses, as expected. Surprisingly, however, we observed that synaptotagmin III is highly enriched in synaptic plasma membranes but not in synaptic vesicles. Synaptotagmin VI was also found to be relatively excluded from synaptic vesicles. Our data suggest that synaptotagmins III and VI perform roles in neurons that are not linked to synaptic vesicle exocytosis but to other Ca2+-related nerve terminal events, indicating that the functions of synaptotagmins are more diverse than originally thought. PMID- 10373433 TI - Thermostable flap endonuclease from the archaeon, Pyrococcus horikoshii, cleaves the replication fork-like structure endo/exonucleolytically. AB - The flap endonuclease gene homologue from the hyperthermophilic archaeon, Pyrococcus horikoshii, was overexpressed in Escherichia coli and purified. The results of gel filtration indicated that this protein was a 41-kDa monomer. P. horikoshii flap endonuclease (phFEN) cleaves replication fork-like substrates (RF) and 5' double-strand flap structures (DF) using both flap endonuclease and 5'-3'-exonuclease activities. The mammalian flap endonuclease (mFEN) is a single strand flap-specific endonuclease (Harrington, J. J., and Lieber, M. R. (1994) EMBO J. 13, 1235-1246), but the action patterns of phFEN appear to be quite different from those of mFEN at this point. The DF-specific flap endonuclease and 5'-exonuclease activities have not yet been reported. Therefore, this is the first report of the specific endo/exonuclease activities of phFEN. The DF specific 5'-exonuclease activity degraded the downstream primer of 3' single-flap structure and was 15 times higher than the activities against nicked substrates without 3' flap strand. DF-specific flap endonuclease cleaved the 5' double-flap strand in DF and the lagging strand in RF at the junction portion. Because the RF appears to be the intermediate structure, due to the arrest of the replication fork, the double strand breaks after the arrests of the replication forks are probably caused by phFEN. PMID- 10373434 TI - Modulation of ATPase activity by physical disengagement of the ATP-binding domains of an ABC transporter, the histidine permease. AB - The membrane-bound complex of the prokaryotic histidine permease, a periplasmic protein-dependent ABC transporter, is composed of two hydrophobic subunits, HisQ and HisM, and two identical ATP-binding subunits, HisP, and is energized by ATP hydrolysis. The soluble periplasmic binding protein, HisJ, creates a signal that induces ATP hydrolysis by HisP. The crystal structure of HisP has been resolved and shown to have an "L" shape, with one of its arms (arm I) being involved in ATP binding and the other one (arm II) being proposed to interact with the hydrophobic subunits (Hung, L.-W., Wang, I. X., Nikaido, K., Liu, P.-Q., Ames, G. F.-L., and Kim, S.-H. (1998) Nature 396, 703-707). Here we study the basis for the defect of several HisP mutants that have an altered signaling pathway and hydrolyze ATP constitutively. We use biochemical approaches to show that they produce a loosely assembled membrane complex, in which the mutant HisP subunits are disengaged from HisQ and HisM, suggesting that the residues involved are important in the interaction between HisP and the hydrophobic subunits. In addition, the mutant HisPs are shown to have lower affinity for ADP and to display no cooperativity for ATP. All of the residues affected in these HisP mutants are located in arm II of the crystal structure of HisP, thus supporting the proposed function of arm II of HisP as interacting with HisQ and HisM. A revised model involving a cycle of disengagement and reengagement of HisP is proposed as a general mechanism of action for ABC transporters. PMID- 10373435 TI - Efficient nitroso group transfer from N-nitrosoindoles to nucleotides and 2' deoxyguanosine at physiological pH. A new pathway for N-nitrosocompounds to exert genotoxicity. AB - The endogenous formation of N-nitrosoindoles is of concern since humans are exposed to a variety of naturally occurring and synthetic indolic compounds. As part of a study to evaluate the genotoxicity of N-nitrosoindoles, the reactions of three model compounds with purine nucleotides and 2'-deoxyguanosine at physiological pH were investigated. The profiles of reaction products were identical for each of the N-nitrosoindoles and three distinct pathways of reaction could be discerned. These pathways were: (i) depurination to the corresponding purine bases, (ii) deamination, coupled with depurination, to give hypoxanthine and xanthine, and (iii) formation of the novel nucleotide 2' deoxyoxanosine monophosphate and its corresponding depurination product oxanine in reactions with 2'-deoxyguanosine monophosphate. 2'-Deoxyoxanosine and oxanine were observed in reactions with 2'-deoxyguanosine. Further studies showed that formation of all of these products could be rationalized by an initial transnitrosation step. These results suggest that, in contrast to many other genotoxic N-nitrosocompounds which are known to alkylate DNA, the genotoxicity of N-nitrosoindoles is likely to arise through transfer of the nitroso group to nucleophilic sites on the purine bases. All of the products resulting from transnitrosation by N-nitrosoindoles are potentially mutagenic. These findings reveal a new pathway for N-nitrosocompounds to exert genotoxicity. PMID- 10373436 TI - Relationship between DNA methylation and mutational patterns induced by a sequence selective minor groove methylating agent. AB - Me-lex, a methyl sulfonate ester appended to a neutral N-methylpyrrolecarboxamide based dipeptide, was synthesized to preferentially generate N3-methyladenine (3 MeA) adducts which are expected to be cytotoxic rather than mutagenic DNA lesions. In the present study, the sequence specificity for DNA alkylation by Me lex was determined in the p53 cDNA through the conversion of the adducted sites into single strand breaks and sequencing gel analysis. In order to establish the mutagenic and lethal properties of Me-lex lesions, a yeast expression vector harboring the human wild-type p53 cDNA was treated in vitro with Me-lex, and transfected into a yeast strain containing the ADE2 gene regulated by a p53 responsive promoter. The results showed that: 1) more than 99% of the lesions induced by Me-lex are 3-MeA; 2) the co-addition of distamycin quantitatively inhibited methylation at all minor groove sites; 3) Me-lex selectively methylated A's that are in, or immediately adjacent to, the lex equilibrium binding sites; 4) all but 6 of the 33 independent mutations were base pair substitutions, the majority of which (17/33; 52%) were AT-targeted; 5) AT --> TA transversions were the predominant mutations observed (13/33; 39%); 6) 13 out of 33 (39%) independent mutations involved a single lex-binding site encompassing positions A600-602 and 9 occurred at position 602 which is a real Me-lex mutation hotspot (n = 9, p < 10(-6), Poisson's normal distribution). A hypothetical model for the interpretation of mutational events at this site is proposed. The present work is the first report on mutational properties of Me-lex. Our results suggest that 3 MeA is not only a cytotoxic but also a premutagenic lesion which exerts this unexpected property in a strict sequence-dependent manner. PMID- 10373437 TI - Nicotinic acetylcholine receptors assembled from the alpha7 and beta3 subunits. AB - Intracellular recordings were performed in voltage-clamped Xenopus oocytes upon injection with a mixture of cDNAs encoding the beta3 and mutant alpha7 (L247Talpha7) neuronal nicotinic acetylcholine receptor (nAChR) subunits. The expressed receptors maintained sensitivity to methyllycaconitine and to alpha bungarotoxin but exhibited a functional profile strikingly different from that of the homomeric L247Talpha7 receptor. The heteromeric L247Talpha7beta3 nAChR had a lower apparent affinity and a faster rate of desensitization than L247Talpha7 nAChR, exhibited nonlinearity in the I-V relationship, and was inhibited by 5 hydroxytryptamine, much like wild type alpha7 (WTalpha7) nAChR. Single channel recordings in cell-attached mode revealed unitary events with a slope conductance of 19 picosiemens and a lifetime of 5 ms, both values being much smaller than those of the homomeric receptor channel. Upon injection with a mixture of WTalpha7 and beta3 cDNAs, clear evidence was obtained for the plasma membrane assembly of heteromeric nAChRs, although ACh could not activate these receptors. It is concluded that beta3, long believed to be an orphan subunit, readily co assembles with other subunits to form heteromeric receptors, some of which may be negative regulators of cholinergic function. PMID- 10373438 TI - Functional and physical interaction between WRN helicase and human replication protein A. AB - The human premature aging disorder Werner syndrome (WS) is associated with a large number of symptoms displayed in normal aging. The WRN gene product, a DNA helicase, has been previously shown to unwind short DNA duplexes (33 kilobases of continuous DNA sequence and contains eight exons. The newly discovered first exon, identified by 5'-rapid amplification of cDNA ends, consists of a 5'-untranslated region and is enriched in C+G nucleotides. Two transcription initiation sites starting at the first and at the second exons were determined by primer extension. Molecular characterization shows that alternatively spliced RNA isoforms are generated by the use of two distinct splice acceptor sites in the third exon situated 278 base pairs apart. We determined a partial genomic structure of the human mimecan gene and demonstrated two alternatively spliced RNA transcripts that are generated likewise. Despite the diversity of mimecan transcripts, the primary structure of the core protein is encoded from exons 3 to 8 and remains unchanged, indicating its functional importance. Using ribonuclease protection assay, we analyzed the patterns of spliced RNA expressed in cultured bovine keratocytes. We demonstrated that their expression is differentially modulated in a temporal manner by basic fibroblast growth factor. PMID- 10373483 TI - Tumor necrosis factor alpha-induced pancreatic beta-cell insulin resistance is mediated by nitric oxide and prevented by 15-deoxy-Delta12,14-prostaglandin J2 and aminoguanidine. A role for peroxisome proliferator-activated receptor gamma activation and inos expression. AB - Recent studies have identified a beta-cell insulin receptor that functions in the regulation of protein translation and mitogenic signaling similar to that described for insulin-sensitive cells. These findings have raised the novel possibility that beta-cells may exhibit insulin resistance similar to skeletal muscle, liver, and fat. To test this hypothesis, the effects of tumor necrosis factor-alpha (TNFalpha), a cytokine proposed to mediate insulin resistance by interfering with insulin signaling at the level of the insulin receptor and its substrates, was evaluated. TNFalpha inhibited p70(s6k) activation by glucose stimulated beta-cells of the islets of Langerhans in a dose- and time-dependent manner, with maximal inhibition observed at approximately 20-50 ng/ml, detected after 24 and 48 h of exposure. Exogenous insulin failed to prevent TNFalpha induced inhibition of p70(s6k), suggesting a defect in the insulin signaling pathway. To further define mechanisms responsible for this inhibition and also to exclude cytokine-induced nitric oxide (NO) as a mediator, the ability of exogenous or endogenous insulin +/- inhibitors of nitric-oxide synthase (NOS) activity, aminoguanidine or N-monomethyl-L-arginine, was evaluated. Unexpectedly, TNFalpha and also interleukin 1 (IL-1)-induced inhibition of p70(s6k) was completely prevented by inhibitors that block NO production. Western blot analysis verified inducible NOS (iNOS) expression after TNFalpha exposure. Furthermore, the ability of IL-1 receptor antagonist protein, IRAP, to block TNFalpha-induced inhibition of p70(s6k) indicated that activation of intra-islet macrophages and the release of IL-1 that induces iNOS expression in beta-cells was responsible for the inhibitory effects of TNFalpha. This mechanism was confirmed by the ability of the peroxisome proliferator-activated receptor-gamma agonist 15-deoxy-Delta12, 14-prostaglandin J2 to attenuate TNFalpha-induced insulin resistance by down-regulating iNOS expression and/or blocking IL-1 release from activated macrophages. Overall, TNFalpha-mediated insulin resistance in beta-cells is characterized by a global inhibition of metabolism mediated by NO differing from that proposed for this proinflammatory cytokine in insulin sensitive cells. PMID- 10373484 TI - p73 and p63 are homotetramers capable of weak heterotypic interactions with each other but not with p53. AB - Mutations in the p53 tumor suppressor gene are the most frequent genetic alterations found in human cancers. Recent identification of two human homologues of p53 has raised the prospect of functional interactions between family members via a conserved oligomerization domain. Here we report in vitro and in vivo analysis of homo- and hetero-oligomerization of p53 and its homologues, p63 and p73. The oligomerization domains of p63 and p73 can independently fold into stable homotetramers, as previously observed for p53. However, the oligomerization domain of p53 does not associate with that of either p73 or p63, even when p53 is in 15-fold excess. On the other hand, the oligomerization domains of p63 and p73 are able to weakly associate with one another in vitro. In vivo co-transfection assays of the ability of p53 and its homologues to activate reporter genes showed that a DNA-binding mutant of p53 was not able to act in a dominant negative manner over wild-type p73 or p63 but that a p73 mutant could inhibit the activity of wild-type p63. These data suggest that mutant p53 in cancer cells will not interact with endogenous or exogenous p63 or p73 via their respective oligomerization domains. It also establishes that the multiple isoforms of p63 as well as those of p73 are capable of interacting via their common oligomerization domain. PMID- 10373485 TI - Histone H1 dephosphorylation is mediated through a radiation-induced signal transduction pathway dependent on ATM. AB - Ionizing radiation is known to activate multiple signal transduction pathways, but the targets of these pathways are poorly understood. Phosphorylation of histone H1 is thought to have a role in chromatin condensation/decondensation, and we asked whether ionizing radiation (IR) would alter H1 phosphorylation. Our data demonstrate that low doses of IR result in a dramatic, but transient, dephosphorylation of H1 isoforms. The in vivo IR-induced dephosphorylation of H1 is completely blocked by wortmannin and is abrogated in ataxia telangiectasia cells. Furthermore, we measured radiation-induced inhibition of cyclin dependent kinase activity and activation of histone H1 phosphatase activity. Both activities were affected by radiation-induced signals in an ATM-dependent manner. Thus, the rapid IR-induced dephosphorylation of H1 involves a pathway including ATM and a wortmannin-sensitive step leading to both inhibition of cyclin dependent kinase activities as well as activation of H1 phosphatase(s). PMID- 10373486 TI - The membrane-spanning domains of caveolins-1 and -2 mediate the formation of caveolin hetero-oligomers. Implications for the assembly of caveolae membranes in vivo. AB - The mammalian caveolin gene family consists of caveolins-1, -2, and -3. The expression of caveolin-3 is muscle-specific. In contrast, caveolins-1 and -2 are co-expressed, and they form a hetero-oligomeric complex in many cell types, with particularly high levels in adipocytes, endothelial cells, and fibroblasts. These caveolin hetero-oligomers are thought to represent the functional assembly units that drive caveolae formation in vivo. Here, we investigate the mechanism by which caveolins-1 and -2 form hetero-oligomers. We reconstituted this reciprocal interaction in vivo and in vitro using a variety of complementary approaches, including the generation of glutathione S-transferase fusion proteins and synthetic peptides. Taken together, our results indicate that the membrane spanning domains of both caveolins-1 and -2 play a critical role in mediating their ability to interact with each other. This is the first demonstration that these unusual membrane-spanning regions found in the caveolin family play a specific role in protein-protein interactions. PMID- 10373487 TI - Selenium metabolism in Drosophila. Characterization of the selenocysteine tRNA population. AB - The selenocysteine (Sec) tRNA population in Drosophila melanogaster is aminoacylated with serine, forms selenocysteyl-tRNA, and decodes UGA. The Km of Sec tRNA and serine tRNA for seryl-tRNA synthetase is 6.67 and 9.45 nM, respectively. Two major bands of Sec tRNA were resolved by gel electrophoresis. Both tRNAs were sequenced, and their primary structures were indistinguishable and colinear with that of the corresponding single copy gene. They are 90 nucleotides in length and contain three modified nucleosides, 5 methylcarboxymethyluridine, N6-isopentenyladenosine, and pseudouridine, at positions 34, 37, and 55, respectively. Neither form contains 1-methyladenosine at position 58 or 5-methylcarboxymethyl-2'-O-methyluridine, which are characteristically found in Sec tRNA of higher animals. We conclude that the primary structures of the two bands of Sec tRNA resolved by electrophoresis are indistinguishable by the techniques employed and that Sec tRNAs in Drosophila may exist in different conformational forms. The Sec tRNA gene maps to a single locus on chromosome 2 at position 47E or F. To our knowledge, Drosophila is the lowest eukaryote in which the Sec tRNA population has been characterized to date. PMID- 10373488 TI - Insertion of atToc34 into the chloroplastic outer membrane is assisted by at least two proteinaceous components in the import system. AB - Toc34 is a member of the outer membrane translocon complex that mediates the initial stage of protein import into chloroplasts. Toc34, like most outer membrane proteins, is synthesized in the cytosol at its mature size without a cleavable transit peptide. The majority of outer membrane proteins do not require thermolysin-sensitive components on the chloroplastic surface or ATP for their insertion into the outer membrane. However, different results have been obtained concerning the factors required for Toc34 insertion into the outer membrane. Using an Arabidopsis homologue of pea Toc34, atToc34, we show that the insertion of atToc34 was greatly reduced by thermolysin pretreatment of chloroplasts as assayed either by protease digestion or by alkaline extraction. The insertion was also dependent on the presence of ATP or GTP. A mutant of atToc34 with the GTP binding domain deleted still required ATP for optimal insertion, indicating that ATP was used by other protein components in the import system. The ATP-supported insertion was observed even in thermolysin-pretreated chloroplasts, suggesting that the protein component responsible for ATP-stimulated insertion is a different protein from the thermolysin-sensitive component that assists atToc34 insertion. PMID- 10373489 TI - Identification of the plasma membrane H+-biotin symporter of Saccharomyces cerevisiae by rescue of a fatty acid-auxotrophic mutant. AB - Bakers' yeast is auxotrophic for biotin (vitamin H) and depends on the efficient uptake of this compound from the environment. A mutant strain with strongly reduced biotin uptake and with reduced levels of protein biotinylation was identified. The strain was auxotrophic for long-chain fatty acids, and this auxotrophy could be suppressed with high levels of biotin in the medium. After transformation of this mutant with a yeast genomic library, the unassigned open reading frame YGR065C was identified to complement this mutation. This gene codes for a protein with 593 amino acids and 12 putative transmembrane helices. Northern blot analysis revealed that, in wild-type cells, the corresponding mRNA levels were increased at low biotin concentrations. Likewise, cellular biotin uptake was increased with decreasing biotin availability. Expression of YGR065C under the control of the constitutive ADH1 promoter resulted in very high biotin transport rates across the plasma membrane that were no longer regulated by the biotin concentration in the growth medium. We conclude that YGR065C encodes the first biotin transporter identified for a non-mammalian organism and designate this gene VHT1 for vitamin H transporter 1. PMID- 10373490 TI - The fenpropimorph resistance gene FEN2 from Saccharomyces cerevisiae encodes a plasma membrane H+-pantothenate symporter. AB - The product of the FEN2 gene of Saccharomyces cerevisiae has previously been described as a protein conferring sensitivity to the antifungal agent fenpropimorph. Fen2p was postulated to act as a common regulator of carbon and nitrogen catabolite repression and of amino acid and ergosterol biosynthesis. In this paper, we present experimental evidence characterizing Fen2p as a plasma membrane-localized transporter for the vitamin pantothenate. The high affinity transport system (Km = 3.5 microM) is sensitive to uncouplers, suggesting a H+ pantothenate cotransport. Pantothenate transport rates in yeast are modulated by extracellular pantothenate, being maximal at low pantothenate concentrations. It is demonstrated that beta-alanine can suppress the growth defect of FEN2 wild type and fen2 mutant cells on pantothenate-free medium. Evidence is presented that beta-alanine is transported by the general amino acid permease Gap1p. The relation among pantothenate transport, nitrogen catabolite repression, and sensitivity to the antifungal agent fenpropimorph is discussed. PMID- 10373491 TI - Evidence for repressional role of an inverted CCAAT box in cell cycle-dependent transcription of the human DNA topoisomerase IIalpha gene. AB - Expression of DNA topoisomerase IIalpha (topo IIalpha) is cell cycle-regulated at both the transcriptional and the post-transcriptional levels. In order to identify cis-acting elements responsible for transcriptional regulation during the cell cycle, we investigated NIH/3T3 cells stably transfected with luciferase reporter plasmids containing various lengths of the human topo IIalpha gene promoter. Serum-deprived cells expressed low levels of luciferase, and following serum-induced cell cycle re-entry luciferase levels were gradually elevated 2 fold. During S phase, a steep 3-fold increase in luciferase activity was seen, reaching its maximum approximately 22 h after serum addition. This pattern was observed with both a full-length (nucleotides (nt) -295 to +90] and a deletion (nt -90 to +90) promoter construct. In contrast, when testing a deletion construct (nt -51 to +90) lacking the first inverted CCAAT box (ICB1) the S phase specific induction was absent. Mutation of ICB1 revealed that it had a repressive character, since luciferase levels rose rapidly to maximal levels immediately following serum addition. Furthermore, electrophoretic mobility shift assays demonstrated a marked decrease in ICB1 binding activity following serum addition. Together, this suggests a role of ICB1 in cell cycle-dependent repression of topo IIalpha transcription. PMID- 10373492 TI - Order of assembly of human DNA repair excision nuclease. AB - Human excision nuclease removes DNA damage by concerted dual incisions bracketing the lesion. The dual incisions are accomplished by sequential and partly overlapping actions of six repair factors, RPA, XPA, XPC, TFIIH, XPG, and XPF.ERCC1. Of these, RPA, XPA, and XPC have specific binding affinity for damaged DNA. To learn about the role of these three proteins in damage recognition and the order of assembly of the excision nuclease, we measured the binding affinities of XPA, RPA, and XPC to a DNA fragment containing a single (6-4) photoproduct and determined the rate of damage excision under a variety of reaction conditions. We found that XPC has the highest affinity to DNA and that RPA has the highest selectivity for damaged DNA. Under experimental conditions conducive to binding of either XPA + RPA or XPC to damaged DNA, the rate of damage removal was about 5-fold faster for reactions in which XPA + RPA was the first damage recognition factor presented to DNA compared with reactions in which XPC was the first protein that had the opportunity to bind to DNA. We conclude that RPA and XPA are the initial damage sensing factors of human excision nuclease. PMID- 10373493 TI - Analysis of tyrosine phosphorylation-dependent interactions between stimulatory effector proteins and the B cell co-receptor CD22. AB - The B cell-restricted transmembrane glycoprotein CD22 is rapidly phosphorylated on tyrosine in response to cross-linking of the B cell antigen receptor, thereby generating phosphotyrosine motifs in the cytoplasmic domain which recruit intracellular effector proteins that contain Src homology 2 domains. By virtue of its interaction with these effector proteins CD22 modulates signal transduction through the B cell antigen receptor. To define further the molecular mechanism by which CD22 mediates its co-receptor function, phosphopeptide mapping experiments were conducted to determine which of the six tyrosine residues in the cytoplasmic domain are involved in recruitment of the stimulatory effector proteins phospholipase Cgamma (PLCgamma), phosphoinositide 3-kinase (PI3K), Grb2, and Syk. The results obtained indicate that the protein tyrosine kinase Syk interacts with multiple CD22-derived phosphopeptides in both immunoprecipitation and reverse Far Western assays. In contrast, the Grb2.Sos complex was observed to bind exclusively to the fourth phosphotyrosine motif (Y828ENV) from CD22 and does so via a direct interaction based on Far Western and reverse Far Western blotting. Although both PLCgamma and PI3K were observed to bind to multiple phosphopeptides in precipitation experiments, subsequent studies using reverse Far Western blot analysis demonstrated that only the carboxyl-terminal phosphopeptide of CD22 (Y863VTL) binds directly to either one. This finding suggests that PLCgamma and PI3K may be recruited to CD22 either through a direct interaction with Tyr863 or indirectly through an association with one or more intermediate proteins. PMID- 10373494 TI - Metalloendopeptidase EC 3.4.24.15 is necessary for Alzheimer's amyloid-beta peptide degradation. AB - We have investigated the functional relationship between metalloendopeptidase EC 3.4.24.15 (MP24.15) and the amyloid precursor protein involved in Alzheimer's disease (AD) and discovered that the enzyme promotes Abeta degradation. We show here that conditioned medium (CM) of MP24.15 antisense-transfected SKNMC neuroblastoma has significantly higher levels of Abeta. Furthermore, synthetic Abeta degradation was increased or decreased following incubation with CM of sense or antisense-transfected cells, respectively. Soluble Abeta1-42 was degraded more slowly than soluble Abeta1-40, while aggregated Abeta1-42 showed almost no degradation. Pretreatment of CM with serine proteinase inhibitors 4-(2 aminoethyl)benzenesulfonyl fluoride and diisopropyl fluorophosphate completely inhibited Abeta degradation. Additionally, alpha1-antichymotrypsin (ACT), a serpin family inhibitor tightly associated with plaques and elevated in AD brain, blocked up to 60% of Abeta degradation. Interestingly, incubation of CM of MP24. 15-overexpressing cells with ACT formed an SDS-resistant ACT complex, suggesting an ACT-serine proteinase interaction. Recombinant MP24. 15 alone did not degrade Abeta. 14C-Diisopropyl fluorophosphate-radiolabeled CM from MP24.15 overexpressing cells contained increased levels of several active serine proteinases, suggesting that MP24.15 activates one or more Abeta-degrading serine proteases. Thus, ACT may cause Abeta accumulation by inhibiting an Abeta degrading enzyme or by direct binding to Abeta, rendering it degradation resistant. Identification of the Abeta-degrading enzyme and MP24.15's role in its activation is underway. Pharmacological modulation of either enzyme may provide a means of regulating Abeta in the brain. PMID- 10373495 TI - Identification of HHR23A as a substrate for E6-associated protein-mediated ubiquitination. AB - The human papilloma virus E6-associated protein (E6AP) functions as a ubiquitin protein ligase (E3) in the E6-mediated ubiquitination of p53. E6AP is also an E3 in the absence of E6, but its normal cellular substrates have not yet been identified. Here we report the identification of HHR23A, one of the human homologues of the yeast DNA repair protein Rad23, as an E6-independent target of E6AP. HHR23A binds E6AP and is ubiquitinated in vitro in an E6AP-dependent manner. Ubiquitinated forms of endogenous HHR23A are detectable in mammalian cells. Overexpression of wild-type E6AP in vivo enhances the ubiquitination of HHR23A, whereas a dominant negative E6AP mutant inhibits HHR23A ubiquitination. Although HHR23A is a stable protein in non-synchronized cells, its levels are regulated in a cell cycle-dependent manner, with specific degradation occurring during S phase. The S phase degradation of HHR23A could be blocked in vivo by dominant negative E6AP, providing direct evidence for the involvement of E6AP in the regulation of HHR23A. Consistent with a role of the HHR23 proteins in DNA repair, UV-induced DNA damage inhibited HHR23A degradation. Although the precise role of HHR23 proteins in DNA repair and cell cycle progression remains to be elucidated, our data suggest that E6AP-mediated ubiquitination of HHR23A may have important implications in DNA repair and cell cycle progression. PMID- 10373496 TI - Gbetagamma and palmitate target newly synthesized Galphaz to the plasma membrane. AB - The subcellular location of a signaling protein determines its ability to transmit messages accurately and efficiently. Three different lipid modifications tether heterotrimeric G proteins to membranes: alpha subunits are myristoylated and/or palmitoylated, and gamma subunits are prenylated. In a previous study, we examined the role of lipid modifications in maintaining the membrane attachment of a G protein alpha subunit, alphaz, which is myristoylated and palmitoylated (Morales, J., Fishburn, C. S., Wilson, P. T., and Bourne, H. R. (1998) Mol. Biol. Cell 9, 1-14). Now we extend this analysis by characterizing the mechanisms that target newly synthesized alphaz to the plasma membrane (PM) and analyze the role of lipid modifications in this process. In comparison with newly synthesized alphas, which is palmitoylated but not myristoylated, alphaz moves more rapidly to the membrane fraction following synthesis in the cytosol. Newly synthesized alphaz associates randomly with cellular membranes, but with time accumulates at the PM. Palmitoylated alphaz is present only in PM-enriched fractions, whereas a nonpalmitoylated mutant of alphaz (alphazC3A) associates less stably with the PM than does wild-type alphaz. Expression of a C-terminal fragment of the beta adrenoreceptor kinase, which sequesters free betagamma, impairs association of both alphaz and alphazC3A with the PM, suggesting that the alpha subunit must bind betagamma in order to localize at the PM. Based on these findings, we propose a model in which, following synthesis on soluble ribosomes, myristoylated alphaz associates randomly and reversibly with membranes; upon association with the PM, alphaz binds betagamma, which promotes its palmitoylation, thus securing it in the proper place for transmitting the hormonal signal. PMID- 10373497 TI - Distribution and fluidizing action of soluble and aggregated amyloid beta-peptide in rat synaptic plasma membranes. AB - The effects of soluble and aggregated amyloid beta-peptide (Abeta) on cortical synaptic plasma membrane (SPM) structure were examined using small angle x-ray diffraction and fluorescence spectroscopy approaches. Electron density profiles generated from the x-ray diffraction data demonstrated that soluble and aggregated Abeta1-40 peptides associated with distinct regions of the SPM. The width of the SPM samples, including surface hydration, was 84 A at 10 degrees C. Following addition of soluble Abeta1-40, there was a broad increase in electron density in the SPM hydrocarbon core +/-0-15 A from the membrane center, and a reduction in hydrocarbon core width by 6 A. By contrast, aggregated Abeta1-40 contributed electron density to the phospholipid headgroup/hydrated surface of the SPM +/-24-37 A from the membrane center, concomitant with an increase in molecular volume in the hydrocarbon core. The SPM interactions observed for Abeta1-40 were reproduced in a brain lipid membrane system. In contrast to Abeta1 40, aggregated Abeta1-42 intercalated into the lipid bilayer hydrocarbon core +/ 0-12 A from the membrane center. Fluorescence experiments showed that both soluble and aggregated Abeta1-40 significantly increased SPM bulk and protein annular fluidity. Physico-chemical interactions of Abeta with the neuronal membrane may contribute to mechanisms of neurotoxicity, independent of specific receptor binding. PMID- 10373498 TI - BRCA1 expression restores radiation resistance in BRCA1-defective cancer cells through enhancement of transcription-coupled DNA repair. AB - The breast cancer predisposition genes, BRCA1 and BRCA2, are responsible for the vast majority of hereditary breast cancer. Although BRCA2 functions to help the cell repair double-stranded DNA breaks, the function of BRCA1 remains enigmatic. Here, we develop a human genetic system to study the role of BRCA1 in oxidative DNA damage. We show that human cancer cells containing mutated BRCA1 are hypersensitive to ionizing radiation. This hypersensitivity can be reversed by the expression of forms of BRCA1 that are not growth suppressing. Reversal of hypersensitivity requires the ring finger of BRCA1, its transactivation domain, and its BRCT domain. Lastly, we show that unlike BRCA2, BRCA1 does not function in the repair of double-stranded DNA breaks. Instead, it functions in transcription-coupled DNA repair (TCR). TCR ability correlated with radioresistance as cells containing BRCA1 showed both increased TCR and radioresistance, whereas cells without BRCA1 showed decreased TCR and radiosensitivity. These findings give physiologic significance to the interaction of BRCA1 with the basal transcription machinery. PMID- 10373499 TI - cGMP binding to noncatalytic sites on mammalian rod photoreceptor phosphodiesterase is regulated by binding of its gamma and delta subunits. AB - The binding of cGMP to the noncatalytic sites on two isoforms of the phosphodiesterase (PDE) from mammalian rod outer segments has been characterized to evaluate their role in regulating PDE during phototransduction. Nonactivated, membrane-associated PDE (PDE-M, alpha beta gamma2) has one exchangeable site for cGMP binding; endogenous cGMP remains nonexchangeable at the second site. Non activated, soluble PDE (PDE-S, alpha beta gamma2 delta) can release and bind cGMP at both noncatalytic sites; the delta subunit is likely responsible for this difference in cGMP exchange rates. Removal of the delta and/or gamma subunits yields a catalytic alphabeta dimer with identical catalytic and binding properties for both PDE-M and PDE-S as follows: high affinity cGMP binding is abolished at one site (KD >1 microM); cGMP binding affinity at the second site (KD approximately 60 nM) is reduced 3-4-fold compared with the nonactivated enzyme; the kinetics of cGMP exchange to activated PDE-M and PDE-S are accelerated to similar extents. The properties of nonactivated PDE can be restored upon addition of gamma subunit. Occupancy of the noncatalytic sites by cGMP may modulate the interaction of the gamma subunit with the alphabeta dimer and thereby regulate cytoplasmic cGMP concentration and the lifetime of activated PDE during visual transduction in photoreceptor cells. PMID- 10373500 TI - ADAMTS-1 is an active metalloproteinase associated with the extracellular matrix. AB - Cellular disintegrin and metalloproteinases (ADAMs) are a family of genes with a sequence similar to the snake venom metalloproteinases and disintegrins. ADAMTS-1 is a unique ADAM family protein with respect to the presence of thrombospondin type I motifs and the capacity to bind to the extracellular matrix. Because ADAMTS-1 has a potential zinc-binding motif in the metalloproteinase domain, we examined in this study whether ADAMTS-1 is an active metalloproteinase by means of the proteinase trapping mechanism of alpha2-macroglobulin. We found that the soluble type of ADAMTS-1 protein is able to form a covalent-binding complex with alpha2-macroglobulin. Furthermore, the point mutation within the zinc-binding motif of ADAMTS-1 protein eliminates its capacity to bind to alpha2 macroglobulin. These data demonstrate that the metalloproteinase domain of ADAMTS 1 is catalytically active. In addition, we showed that the removal of the pro domain from the ADAMTS-1 precursor is impaired in the furin-deficient cell line, LoVo, and that the processing ability of the cells is restored by the co expression of the furin cDNA. These data provide evidence that the ADAMTS-1 precursor is processed in vivo by furin endopeptidase in the secretory pathway. Consequently, ADAMTS-1 is an active metalloprotease that is associated with the extracellular matrix. This study strongly suggests that ADAMTS-1 may play a role in the inflammatory process through its protease activity. PMID- 10373501 TI - Negative regulation of transactivation function but not DNA binding of NF-kappaB and AP-1 by IkappaBbeta1 in breast cancer cells. AB - The transcription factor NF-kappaB regulates the expression of genes involved in cancer cell invasion, metastasis, angiogenesis, and resistance to chemotherapy. In normal cells NF-kappaB is maintained in the cytoplasm by protein-protein interaction with inhibitor IkappaBs. In contrast, in cancer cells a substantial amount of NF-kappaB is in the nucleus and constitutively activates target genes. To understand the mechanisms of constitutive NF-kappaB activation, we have analyzed the function of IkappaBalpha and IkappaBbeta in breast cancer cells. In most cases, constitutive NF-kappaB DNA binding correlated with reduced levels of either IkappaBalpha or IkappaBbeta isoforms. Overexpression of IkappaBalpha but not IkappaBbeta1 resulted in reduced constitutive DNA binding of NF-kappaB in MDA MB-231 cells. Unexpectedly, IkappaBbeta1 overexpression moderately increased 12-O tetradecanoylphorbol-13-acetate- and interleukin-1-inducible NF-kappaB DNA binding. 12-O-Tetradecanoylphorbol-13-acetate- and interleukin-1-induced transactivation by NF-kappaB, however, was lower in IkappaBbeta1-overexpressing cells. Mutants of IkappaBbeta1 lacking the C-terminal casein kinase II phosphorylation sites, which form a stable complex with DNA bound NF-kappaB without inhibiting its transactivation in other cell types, repressed the transactivation by NF-kappaB in MDA-MB-231 cells. Consistent with the results of transient transfections, the expression of urokinase plasminogen activator, an NF kappaB target gene, was reduced in IkappaBbeta1-overexpressing cells. These results suggest that depending on the cell type, IkappaBbeta1 represses the expression of NF-kappaB-regulated genes by inhibiting either DNA binding or transactivation function of NF-kappaB. PMID- 10373502 TI - Amino-terminal cysteine residues of RGS16 are required for palmitoylation and modulation of Gi- and Gq-mediated signaling. AB - RGS proteins (Regulators of G protein Signaling) are a recently discovered family of proteins that accelerate the GTPase activity of heterotrimeric G protein alpha subunits of the i, q, and 12 classes. The proteins share a homologous core domain but have divergent amino-terminal sequences that are the site of palmitoylation for RGS-GAIP and RGS4. We investigated the function of palmitoylation for RGS16, which shares conserved amino-terminal cysteines with RGS4 and RGS5. Mutation of cysteine residues at residues 2 and 12 blocked the incorporation of [3H]palmitate into RGS16 in metabolic labeling studies of transfected cells or into purified RGS proteins in a cell-free palmitoylation assay. The purified RGS16 proteins with the cysteine mutations were still able to act as GTPase-activating protein for Gialpha. Inhibition or a decrease in palmitoylation did not significantly change the amount of protein that was membrane-associated. However, palmitoylation-defective RGS16 mutants demonstrated impaired ability to inhibit both Gi- and Gq-linked signaling pathways when expressed in HEK293T cells. These findings suggest that the amino-terminal region of RGS16 may affect the affinity of these proteins for Galpha subunits in vivo or that palmitoylation localizes the RGS protein in close proximity to Galpha subunits on cellular membranes. PMID- 10373503 TI - Bridging the gap: composition, regulation, and physiological function of the IkappaB kinase complex. PMID- 10373504 TI - An mRNA stability complex functions with poly(A)-binding protein to stabilize mRNA in vitro. AB - The stable globin mRNAs provide an ideal system for studying the mechanism governing mammalian mRNA turnover. alpha-Globin mRNA stability is dictated by sequences in the 3' untranslated region (3'UTR) which form a specific ribonucleoprotein complex (alpha-complex) whose presence correlates with mRNA stability. One of the major protein components within this complex is a family of two polycytidylate-binding proteins, alphaCP1 and alphaCP2. Using an in vitro transcribed and polyadenylated alpha-globin 3'UTR, we have devised an in vitro mRNA decay assay which reproduces the alpha-complex-dependent mRNA stability observed in cells. Incubation of the RNA with erythroleukemia K562 cytosolic extract results in deadenylation with distinct intermediates containing a periodicity of approximately 30 nucleotides, which is consistent with the binding of poly(A)-binding protein (PABP) monomers. Disruption of the alpha-complex by sequestration of alphaCP1 and alphaCP2 enhances deadenylation and decay of the mRNA, while reconstitution of the alpha-complex stabilizes the mRNA. Similarly, PABP is also essential for the stability of mRNA in vitro, since rapid deadenylation resulted upon its depletion. An RNA-dependent interaction between alphaCP1 and alphaCP2 with PABP suggests that the alpha-complex can directly interact with PABP. Therefore, the alpha-complex is an mRNA stability complex in vitro which could function at least in part by interacting with PABP. PMID- 10373505 TI - Std1 and Mth1 proteins interact with the glucose sensors to control glucose regulated gene expression in Saccharomyces cerevisiae. AB - The Std1 protein modulates the expression of glucose-regulated genes, but its exact molecular role in this process is unclear. A two-hybrid screen for Std1 interacting proteins identified the hydrophilic C-terminal domains of the glucose sensors, Snf3 and Rgt2. The homologue of Std1, Mth1, behaves differently from Std1 in this assay by interacting with Snf3 but not Rgt2. Genetic interactions between STD1, MTH1, SNF3, and RGT2 suggest that the glucose signaling is mediated, at least in part, through interactions of the products of these four genes. Mutations in MTH1 can suppress the raffinose growth defect of a snf3 mutant as well as the glucose fermentation defect present in cells lacking both glucose sensors (snf3 rgt2). Genetic suppression by mutations in MTH1 is likely to be due to the increased and unregulated expression of hexose transporter genes. In media lacking glucose or with low levels of glucose, the hexose transporter genes are subject to repression by a mechanism that requires the Std1 and Mth1 proteins. An additional mechanism for glucose sensing must exist since a strain lacking all four genes (snf3 rgt2 std1 mth1) is still able to regulate SUC2 gene expression in response to changes in glucose concentration. Finally, studies with green fluorescent protein fusions indicate that Std1 is localized to the cell periphery and the cell nucleus, supporting the idea that it may transduce signals from the plasma membrane to the nucleus. PMID- 10373506 TI - Assembly of the alpha-globin mRNA stability complex reflects binary interaction between the pyrimidine-rich 3' untranslated region determinant and poly(C) binding protein alphaCP. AB - Globin mRNAs accumulate to 95% of total cellular mRNA during terminal erythroid differentiation, reflecting their extraordinary stability. The stability of human alpha-globin mRNA is paralleled by formation of a sequence-specific RNA-protein (RNP) complex at a pyrimidine-rich site within its 3' untranslated region (3'UTR), the alpha-complex. The proteins of the alpha-complex are widely expressed. The alpha-complex or a closely related complex also assembles at pyrimidine-rich 3'UTR segments of other stable mRNAs. These data suggest that the alpha-complex may constitute a general determinant of mRNA stability. One or more alphaCPs, members of a family of hnRNP K-homology domain poly(C) binding proteins, are essential constituents of the alpha-complex. The ability of alphaCPs to homodimerize and their reported association with additional RNA binding proteins such as AU-rich binding factor 1 (AUF1) and hnRNP K have suggested that the alpha-complex is a multisubunit structure. In the present study, we have addressed the composition of the alpha-complex. An RNA titration recruitment assay revealed that alphaCPs were quantitatively incorporated into the alpha-complex in the absence of associated AUF1 and hnRNP K. A high-affinity direct interaction between each of the three major alphaCP isoforms and the alpha globin 3'UTR was detected, suggesting that each of these proteins might be sufficient for alpha-complex assembly. This sufficiency was further supported by the sequence-specific binding of recombinant alphaCPs to a spectrum of RNA targets. Finally, density sedimentation analysis demonstrated that the alpha complex could accommodate only a single alphaCP. These data established that a single alphaCP molecule binds directly to the alpha-globin 3'UTR, resulting in a simple binary structure for the alpha-complex. PMID- 10373507 TI - Dominant negative murine serum response factor: alternative splicing within the activation domain inhibits transactivation of serum response factor binding targets. AB - Primary transcripts encoding the MADS box superfamily of proteins, such as MEF2 in animals and ZEMa in plants, are alternatively spliced, producing several isoformic species. We show here that murine serum response factor (SRF) primary RNA transcripts are alternatively spliced at the fifth exon, deleting approximately one-third of the C-terminal activation domain. Among the different muscle types examined, visceral smooth muscles have a very low ratio of SRFDelta5 to SRF. Increased levels of SRFDelta5 correlates well with reduced smooth muscle contractile gene activity within the elastic aortic arch, suggesting important biological roles for differential expression of SRFDelta5 variant relative to wild-type SRF. SRFDelta5 forms DNA binding-competent homodimers and heterodimers. SRFDelta5 acts as a naturally occurring dominant negative regulatory mutant that blocks SRF-dependent skeletal alpha-actin, cardiac alpha-actin, smooth alpha actin, SM22alpha, and SRF promoter-luciferase reporter activities. Expression of SRFDelta5 interferes with differentiation of myogenic C2C12 cells and the appearance of skeletal alpha-actin and myogenin mRNAs. SRFDelta5 repressed the serum-induced activity of the c-fos serum response element. SRFDelta5 fused to the yeast Gal4 DNA binding domain displayed low transcriptional activity, which was complemented by overexpression of the coactivator ATF6. These results indicate that the absence of exon 5 might be bypassed through recruitment of transcription factors that interact with extra-exon 5 regions in the transcriptional activating domain. The novel alternatively spliced isoform of SRF, SRFDelta5, may play an important regulatory role in modulating SRF-dependent gene expression. PMID- 10373508 TI - Highly divergent lentiviral Tat proteins activate viral gene expression by a common mechanism. AB - The human immunodeficiency virus type 1 (HIV-1) Tat protein (hTat) activates transcription initiated at the viral long terminal repeat (LTR) promoter by a unique mechanism requiring recruitment of the human cyclin T1 (hCycT1) cofactor to the viral TAR RNA target element. While activation of equine infectious anemia virus (EIAV) gene expression by the EIAV Tat (eTat) protein appears similar in that the target element is a promoter proximal RNA, eTat shows little sequence homology to hTat, does not activate the HIV-1 LTR, and is not active in human cells that effectively support hTat function. To address whether eTat and hTat utilize similar or distinct mechanisms of action, we have cloned the equine homolog of hCycT1 (eCycT1) and examined whether it is required to mediate eTat function. Here, we report that expression of eCycT1 in human cells fully rescues eTat function and that eCycT1 and eTat form a protein complex that specifically binds to the EIAV, but not the HIV-1, TAR element. While hCycT1 is also shown to interact with eTat, the lack of eTat function in human cells is explained by the failure of the resultant protein complex to bind to EIAV TAR. Critical sequences in eCycT1 required to support eTat function are located very close to the amino terminus, i.e., distal to the HIV-1 Tat-TAR interaction motif previously identified in the hCycT1 protein. Together, these data provide a molecular explanation for the species tropism displayed by eTat and demonstrate that highly divergent lentiviral Tat proteins activate transcription from their cognate LTR promoters by essentially identical mechanisms. PMID- 10373509 TI - Target specificities of Drosophila enhancer of split basic helix-loop-helix proteins. AB - Seven Enhancer of split genes in Drosophila melanogaster encode basic-helix-loop helix transcription factors which are components of the Notch signalling pathway. They are expressed in response to Notch activation and mediate some effects of the pathway by regulating the expression of target genes. Here we have determined that the optimal DNA binding site for the Enhancer of split proteins is a palindromic 12-bp sequence, 5'-TGGCACGTG(C/T)(C/T)A-3', which contains an E-box core (CACGTG). This site is recognized by all of the individual Enhancer of split basic helix-loop-helix proteins, consistent with their ability to regulate similar target genes in vivo. We demonstrate that the 3 bp flanking the E-box core are intrinsic to DNA recognition by these proteins and that the Enhancer of split and proneural proteins can compete for binding on specific DNA sequences. Furthermore, the regulation conferred on a reporter gene in Drosophila by three closely related sequences demonstrates that even subtle sequence changes within an E box or flanking bases have dramatic consequences on the overall repertoire of proteins that can bind in vivo. PMID- 10373510 TI - Ras-specific exchange factor GRF: oligomerization through its Dbl homology domain and calcium-dependent activation of Raf. AB - The full-length versions of the Ras-specific exchange factors Ras-GRF1 (GRF1) and Ras-GRF2 (GRF2), which are expressed in brain and a restricted number of other organs, possess an ionomycin-dependent activation of Erk mitogen-activated protein kinase activity in 293T cells (C. L. Farnsworth et al., Nature 376:524 527, 1995; N. P. Fam et al., Mol. Cell. Biol. 17:1396-1406, 1996). Each GRF protein contains a Dbl homology (DH) domain. A yeast two-hybrid screen was used to identify polypeptides that associate with the DH domain of GRF1. In this screen, a positive cDNA clone from a human brain cDNA library was isolated which consisted of the GRF2 DH domain and its adjacent ilimaquinone domain. Deletion analysis verified that the two-hybrid interaction required only the DH domains, and mutation of Leu-263 to Gln (L263Q) in the N terminus of the GRF1 DH domain abolished the two-hybrid interaction, while a cluster of more C-terminally located mutations in the DH domain did not eliminate the interaction. Oligomers between GRF1 and GRF2 were detected in a rat brain extract, and forced expression of GRF1 and GRF2 in cultured mammalian cells formed homo- and hetero-oligomers. Introduction of the L263Q mutation in GRF1 led to a protein that was deficient in oligomer formation, while GRF1 containing the DH cluster mutations formed homo oligomers with an efficiency similar to that of wild type. Compared to wild-type GRF1, the focus-forming activity on NIH 3T3 cells of the GRF1 DH cluster mutant was reduced, while the L263Q mutant was inactive. Both mutants were impaired in their ability to mediate ionomycin-dependent Erk activity in 293T cells. In the absence of ionomycin, 293T cells expressing wild-type GRF1 contained much higher levels of Ras-GTP than control cells; the increase in Erk activity induced by ionomycin in the GRF1-expressing cells also induced a concomitant increase in Raf kinase activity, but without a further increase in the level Ras-GTP. We conclude that GRF1 and GRF2 can form homo- and hetero-oligomers via their DH domains, that mutational inactivation of oligomer formation by GRF1 is associated with impaired biological and signaling activities, and that in 293T cells GRF1 mediates at least two pathways for Raf activation: one a constitutive signal that is mainly Ras-dependent, and one an ionomycin-induced signal that cooperates with the constitutive signal without further augmenting the level of GTP-Ras. PMID- 10373511 TI - Cellular ras and cyclin D1 are required during different cell cycle periods in cycling NIH 3T3 cells. AB - Novel techniques were used to determine when in the cell cycle of proliferating NIH 3T3 cells cellular Ras and cyclin D1 are required. For comparison, in quiescent cells, all four of the inhibitors of cell cycle progression tested (anti-Ras, anti-cyclin D1, serum removal, and cycloheximide) became ineffective at essentially the same point in G1 phase, approximately 4 h prior to the beginning of DNA synthesis. To extend these studies to cycling cells, a time lapse approach was used to determine the approximate cell cycle position of individual cells in an asynchronous culture at the time of inhibitor treatment and then to determine the effects of the inhibitor upon recipient cells. With this approach, anti-Ras antibody efficiently inhibited entry into S phase only when introduced into cells prior to the preceding mitosis, several hours before the beginning of S phase. Anti-cyclin D1, on the other hand, was an efficient inhibitor when introduced up until just before the initiation of DNA synthesis. Cycloheximide treatment, like anti-cyclin D1 microinjection, was inhibitory throughout G1 phase (which lasts a total of 4 to 5 h in these cells). Finally, serum removal blocked entry into S phase only during the first hour following mitosis. Kinetic analysis and a novel dual-labeling technique were used to confirm the differences in cell cycle requirements for Ras, cyclin D1, and cycloheximide. These studies demonstrate a fundamental difference in mitogenic signal transduction between quiescent and cycling NIH 3T3 cells and reveal a sequence of signaling events required for cell cycle progression in proliferating NIH 3T3 cells. PMID- 10373512 TI - Interaction between the product of the breast cancer susceptibility gene BRCA2 and DSS1, a protein functionally conserved from yeast to mammals. AB - Germ line mutations in the breast cancer susceptibility gene BRCA2 predispose to early-onset breast cancer, but the function of the nuclear protein encoded by the gene is ill defined. Using the yeast two-hybrid system with fragments of human BRCA2, we identified an interaction with the human DSS1 (deleted in split hand/split foot) gene. Yeast and mammalian two-hybrid assays showed that DSS1 can associate with BRCA2 in the region of amino acids 2472 to 2957 in the C terminus of the protein. Using coimmunoprecipitation of epitope-tagged BRCA2 and DSS1 cDNA constructs transiently expressed in COS cells, we were able to demonstrate an association. Furthermore, endogenous BRCA2 could be coimmunoprecipitated with endogenous DSS1 in MCF7 cells, demonstrating an in vivo association. Apparent orthologues of the mammalian DSS1 gene were identified in the genome of the yeasts Schizosaccharomyces pombe and Saccharomyces cerevisiae. Yeast strains in which these DSS1-like genes were deleted showed a temperature-sensitive growth phenotype, which was analyzed by flow cytometry. This provides evidence for a link between the BRCA2 tumor suppressor gene and a gene required for completion of the cell cycle. PMID- 10373513 TI - Mutant cells that do not respond to interleukin-1 (IL-1) reveal a novel role for IL-1 receptor-associated kinase. AB - Mutagenized human 293 cells containing an interleukin-1 (IL-1)-regulated herpes thymidine kinase gene, selected in IL-1 and gancyclovir, have yielded many independent clones that are unresponsive to IL-1. The four clones analyzed here carry recessive mutations and represent three complementation groups. Mutant A in complementation group I1 lacks IL-1 receptor-associated kinase (IRAK), while the mutants in the other two groups are defective in unknown components that function upstream of IRAK. Expression of exogenous IRAK in I1A cells (I1A-IRAK) restores their responsiveness to IL-1. Neither NFkappaB nor Jun kinase is activated in IL 1-treated I1A cells, but these responses are restored in I1A-IRAK cells, indicating that IRAK is required for both. To address the role of the kinase activity of IRAK in IL-1 signaling, its ATP binding site was mutated (K239A), completely abolishing kinase activity. In transfected I1A cells, IRAK-K239A was still phosphorylated upon IL-1 stimulation and, surprisingly, still complemented all the defects in the mutant cells. Therefore, IRAK must be phosphorylated by a different kinase, and phospho-IRAK must play a role in IL-1-mediated signaling that does not require its kinase activity. PMID- 10373514 TI - Induction of apoptosis by double-stranded-RNA-dependent protein kinase (PKR) involves the alpha subunit of eukaryotic translation initiation factor 2 and NF kappaB. AB - The double-stranded (ds) RNA-dependent protein kinase (PKR) is a key mediator of antiviral effects of interferon (IFN) and an active player in apoptosis induced by different stimuli. The translation initiation factor eIF-2alpha (alpha subunit of eukaryotic translation initiation factor 2) and IkappaBalpha, the inhibitor of the transcription factor NF-kappaB, have been proposed as downstream mediators of PKR effects. To evaluate the involvement of NF-kappaB and eIF-2alpha in the induction of apoptosis by PKR, we have used vaccinia virus (VV) recombinants that inducibly express PKR concomitantly with a dominant negative mutant of eIF-2alpha or a repressor form of IkappaBalpha. We found that while expression of PKR by a VV vector resulted in extensive inhibition of protein synthesis and induction of apoptosis, coexpression of PKR with a dominant negative mutant of eIF-2alpha (Ser 51-->Ala) reversed both the PKR-mediated translational block and PKR-induced apoptosis. Coexpression of PKR with a repressor form of IkappaBalpha (Ser-32, 36 Ala) also leads to the inhibition of apoptosis by abolishing NF-kappaB induction, while translation remains blocked. Treating cells with two different proteasome inhibitors which block IkappaBalpha degradation, prevented PKR-induced apoptosis, supporting results from coexpression studies. Biochemical analysis and transient assays revealed that PKR expression by a VV vector induced NF-kappaB binding and transactivation. In addition, upregulation of Fas mRNA transcription occurred during PKR activation. Our findings provide direct evidence for the involvement of eIF-2alpha and NF-kappaB in the induction of apoptosis by PKR. PMID- 10373515 TI - A RAG1 and RAG2 tetramer complex is active in cleavage in V(D)J recombination. AB - During V(D)J recombination two proteins, RAG1 and RAG2, assemble as a protein-DNA complex with the appropriate DNA targets containing recombination signal sequences (RSSs). The properties of this complex require a fairly elaborate set of protein-protein and protein-DNA contacts. Here we show that a purified derivative of RAG1, without DNA, exists predominantly as a homodimer. A RAG2 derivative alone has monomer, dimer, and larger forms. The coexpressed RAG1 and RAG2 proteins form a mixed tetramer in solution which contains two molecules of each protein. The same tetramer of RAG1 and RAG2 plus one DNA molecule is the form active in cleavage. Additionally, we show that both DNA products following cleavage can still be held together in a stable protein-DNA complex. PMID- 10373517 TI - Osmotic shock inhibits insulin signaling by maintaining Akt/protein kinase B in an inactive dephosphorylated state. AB - We have previously reported that insulin and osmotic shock stimulate an increase in glucose transport activity and translocation of the insulin-responsive glucose transporter isoform GLUT4 to the plasma membrane through distinct pathways in 3T3L1 adipocytes (D. Chen, J. S. Elmendorf, A. L. Olson, X. Li, H. S. Earp, and J. E. Pessin, J. Biol. Chem. 272:27401-27410, 1997). In investigations of the relationships between these two signaling pathways, we have now observed that these two stimuli are not additive, and, in fact, osmotic shock pretreatment was found to completely prevent any further insulin stimulation of glucose transport activity and GLUT4 protein translocation. In addition, osmotic shock inhibited the insulin stimulation of lipogenesis and glycogen synthesis. This inhibition of insulin-stimulated downstream signaling occurred without any significant effect on insulin receptor autophosphorylation or tyrosine phosphorylation of insulin receptor substrate 1 (IRS1). Furthermore, there was no effect on either the insulin-stimulated association of the p85 type I phosphatidylinositol (PI) 3 kinase regulatory subunit with IRS1 or phosphotyrosine antibody immunoprecipitated PI 3-kinase activity. In contrast, osmotic shock pretreatment markedly inhibited the insulin stimulation of protein kinase B (PKB) and p70S6 kinase activities. In addition, the dephosphorylation of PKB was prevented by pretreatment with the phosphatase inhibitors okadaic acid and calyculin A. These data support a model in which osmotic shock-induced insulin resistance of downstream biological responses results from an inhibition of insulin-stimulated PKB activation. PMID- 10373516 TI - c-Myc regulates cyclin D-Cdk4 and -Cdk6 activity but affects cell cycle progression at multiple independent points. AB - c-myc is a cellular proto-oncogene associated with a variety of human cancers and is strongly implicated in the control of cellular proliferation, programmed cell death, and differentiation. We have previously reported the first isolation of a c-myc-null cell line. Loss of c-Myc causes a profound growth defect manifested by the lengthening of both the G1 and G2 phases of the cell cycle. To gain a clearer understanding of the role of c-Myc in cellular proliferation, we have performed a comprehensive analysis of the components that regulate cell cycle progression. The largest defect observed in c-myc-/- cells is a 12-fold reduction in the activity of cyclin D1-Cdk4 and -Cdk6 complexes during the G0-to-S transition. Downstream events, such as activation of cyclin E-Cdk2 and cyclin A-Cdk2 complexes, are delayed and reduced in magnitude. However, it is clear that c-Myc affects the cell cycle at multiple independent points, because restoration of the Cdk4 and -6 defect does not significantly increase growth rate. In exponentially cycling cells the absence of c-Myc reduces coordinately the activities of all cyclin-cyclin-dependent kinase complexes. An analysis of cyclin-dependent kinase complex regulators revealed increased expression of p27(KIP1) and decreased expression of Cdk7 in c-myc-/- cells. We propose that c-Myc functions as a crucial link in the coordinate adjustment of growth rate to environmental conditions. PMID- 10373518 TI - Serum-induced expression of the cdc25A gene by relief of E2F-mediated repression. AB - The cdc25A gene encodes a tyrosine phosphatase which activates cyclin-dependent kinase activity in the G1 phase of the cell cycle. cdc25A RNA levels are induced from 3 to 6 h after serum induction of serum-starved NIH 3T3 cells, suggesting that the cdc25A gene is a delayed-early gene. Analysis of cdc25A promoter constructs showed that the cdc25A promoter is sufficient for serum induction. Surprisingly for a gene expressed in early to mid-G1, serum induction of the promoter requires an E2F site at position -62 in the promoter. Deletion or point mutation of the E2F site resulted in activation of expression in serum-starved cells and no further induction by serum treatment. E2F factors were found to bind to the cdc25A E2F site along with the retinoblastoma protein (Rb) family members p130 and p107. A shift in mobility of the E2F-p107 complex in extracts of cells induced for 6 h correlated with induction of cdc25A expression. These results suggest that serum induction of cdc25A expression is mediated by inactivation of p107 or p130, both of which repress transcription when bound to the promoter through E2F. PMID- 10373519 TI - A Uve1p-mediated mismatch repair pathway in Schizosaccharomyces pombe. AB - UV damage endonuclease (Uve1p) from Schizosaccharomyces pombe was initially described as a DNA repair enzyme specific for the repair of UV light-induced photoproducts and proposed as the initial step in an alternative excision repair pathway. Here we present biochemical and genetic evidence demonstrating that Uve1p is also a mismatch repair endonuclease which recognizes and cleaves DNA 5' to the mispaired base in a strand-specific manner. The biochemical properties of the Uve1p-mediated mismatch endonuclease activity are similar to those of the Uve1p-mediated UV photoproduct endonuclease. Mutants lacking Uve1p display a spontaneous mutator phenotype, further confirming the notion that Uve1p plays a role in mismatch repair. These results suggest that Uve1p has a surprisingly broad substrate specificity and may function as a general type of DNA repair protein with the capacity to initiate mismatch repair in certain organisms. PMID- 10373520 TI - Major Egr3 isoforms are generated via alternate translation start sites and differ in their abilities to activate transcription. AB - In previous studies, we detected a major, unidentified Egr response element (ERE) binding complex in brain extracts. We now report that this complex contains a truncated isoform of Egr3 generated by use of an alternate translation start site at methionine 106. Furthermore, the ERE binding complex previously thought to contain full-length Egr3 includes several isoforms generated by initiation at other internal methionines. Full-length and truncated (missing residues 1 to 105) Egr3 isoforms differ in the ability to stimulate transcription directed by a tandem repeat of two EREs but not by a single ERE. Taken together, our results indicate that alternative translation start sites are used to generate Egr3 isoforms with distinct transcriptional properties. PMID- 10373521 TI - Transcriptional cofactor CA150 regulates RNA polymerase II elongation in a TATA box-dependent manner. AB - Tat protein strongly activates transcription from the human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR) by enhancing the elongation efficiency of RNA polymerase II complexes. Tat-mediated transcriptional activation requires cellular cofactors and specific cis-acting elements within the HIV-1 promoter, among them a functional TATA box. Here, we have investigated the mechanism by which one of these cofactors, termed CA150, regulates HIV-1 transcription in vivo. We present a series of functional assays that demonstrate that the regulation of the HIV-1 LTR by CA150 has the same functional requirements as the activation by Tat. We found that CA150 affects elongation of transcription complexes assembled on the HIV-1 promoter in a TATA-box-dependent manner. We discuss the data in terms of the involvement of CA150 in the regulation of Tat-activated HIV-1 gene expression. In addition, we also provide evidence suggesting a role for CA150 in the regulation of cellular transcriptional processes. PMID- 10373522 TI - p70(s6k) integrates phosphatidylinositol 3-kinase and rapamycin-regulated signals for E2F regulation in T lymphocytes. AB - In T lymphocytes, the hematopoietic cytokine interleukin-2 (IL-2) uses phosphatidylinositol 3-kinase (PI 3-kinase)-induced signaling pathways to regulate E2F transcriptional activity, a critical cell cycle checkpoint. PI 3 kinase also regulates the activity of p70(s6k), the 40S ribosomal protein S6 kinase, a response that is abrogated by the macrolide rapamycin. This immunosuppressive drug is known to prevent T-cell proliferation, but the precise point at which rapamycin regulates T-cell cycle progression has yet to be elucidated. Moreover, the effects of rapamycin on, and the role of p70(s6k) in, IL-2 and PI 3-kinase activation of E2Fs have not been characterized. Our present results show that IL-2- and PI 3-kinase-induced pathways for the regulation of E2F transcriptional activity include both rapamycin-resistant and rapamycin sensitive components. Expression of a rapamycin-resistant mutant of p70(s6k) in T cells could restore rapamycin-suppressed E2F responses. Thus, the rapamycin controlled processes involved in E2F regulation appear to be mediated by p70(s6k). However, the rapamycin-resistant p70(s6k) could not rescue rapamycin inhibition of T-cell cycle entry, consistent with the involvement of additional, rapamycin-sensitive pathways in the control of T-cell cycle progression. The present results thus show that p70(s6k) is able to regulate E2F transcriptional activity and provide direct evidence for the first time for a link between IL-2 receptors, PI 3-kinase, and p70(s6k) that regulates a crucial G1 checkpoint in T lymphocytes. PMID- 10373523 TI - Suppression of E1A-mediated transformation by the p50E4F transcription factor. AB - The adenovirus E1A gene can act as an oncogene or a tumor suppressor, with the latter effect generally arising from the induction of apoptosis or the repression of genes that provide oncogenic growth stimuli (e.g., HER-2/c-erbB2/neu) or increased metastatic invasiveness (e.g., metalloproteases). In this study, coexpression of E1A and p50E4F, a cellular transcription factor whose DNA binding activity is stimulated by E1A, suppressed colony formation by NIH 3T3 cells and transformation of primary rat embryo fibroblasts but had no observed effect in the absence of E1A. Domains in p50E4F required for stimulation of the adenovirus E4 promoter were required for the suppressive effect, indicating a transcriptional mechanism. In serum-containing media, retroviral expression of p50E4F in E1A13S/ras-transformed NIH 3T3 fibroblasts had little effect on subconfluent cultures but accelerated a decline in viability after the cultures reached confluence. Cell death occurred by both apoptosis and necrosis, with the predominance of each process determined by culture conditions. In serum-free media, p50E4F accelerated E1A-induced apoptosis. The results suggest that p50E4F sensitizes cells to signals or conditions that cause cell death. PMID- 10373524 TI - SDF-2 induction of terminal differentiation in Dictyostelium discoideum is mediated by the membrane-spanning sensor kinase DhkA. AB - SDF-2 is a peptide released by prestalk cells during culmination that stimulates prespore cells to encapsulate. Genetic evidence indicates that the response is dependent on the dhkA gene. This gene encodes a member of the histidine kinase family of genes that functions in two-component signal transduction pathways. The sequence of the N-terminal half of DhkA predicts two hydrophobic domains separated by a 310-amino-acid loop that could bind a ligand. By inserting MYC6 epitopes into DhkA, we were able to show that the loop is extracellular while the catalytic domain is cytoplasmic. Cells expressing the MYC epitope in the extracellular domain of DhkA were found to respond only if induced with 100-fold higher levels of SDF-2 than required to induce dhkA+ cells; however, they could be induced to sporulate by addition of antibodies specific to the MYC epitope. To examine the enzymatic activity of DhkA, we purified the catalytic domain following expression in bacteria and observed incorporation of labelled phosphate from ATP consistent with histidine autophosphorylation. Site-directed mutagenesis of histidine1395 to glutamine in the catalytic domain blocked autophosphorylation. Furthermore, genetic analyses showed that histidine1395 and the relay aspartate2075 of DhkA are both critical to its function but that another histidine kinase, DhkB, can partially compensate for the lack of DhkA activity. Sporulation is drastically reduced in double mutants lacking both DhkA and DhkB. Suppressor studies indicate that the cyclic AMP (cAMP) phosphodiesterase RegA and the cAMP-dependent protein kinase PKA act downstream of DhkA. PMID- 10373525 TI - Inhibition of double-stranded RNA- and tumor necrosis factor alpha-mediated apoptosis by tetratricopeptide repeat protein and cochaperone P58(IPK). AB - P58(IPK) is a tetratricopeptide repeat-containing cochaperone that is involved in stress-activated cellular pathways and that inhibits the activity of protein kinase PKR, a primary mediator of the antiviral and antiproliferative properties of interferon. To gain better insight into the molecular actions of P58(IPK), we generated NIH 3T3 cell lines expressing either wild-type P58(IPK) or a P58(IPK) deletion mutant, DeltaTPR6, that does not bind to or inhibit PKR. When treated with double-stranded RNA (dsRNA), DeltaTPR6-expressing cells exhibited a significant increase in eukaryotic initiation factor 2alpha phosphorylation and NF-kappaB activation, indicating a functional PKR. In contrast, both of these PKR dependent events were blocked by the overexpression of wild-type P58(IPK). In addition, the P58(IPK) cell line, but not the DeltaTPR6 cell line, was resistant to dsRNA-induced apoptosis. Together, these findings demonstrate that P58(IPK) regulates dsRNA signaling pathways by inhibiting multiple PKR-dependent functions. In contrast, both the P58(IPK) and DeltaTPR6 cell lines were resistant to tumor necrosis factor alpha-induced apoptosis, suggesting that P58(IPK) may function as a more general suppressor of programmed cell death independently of its PKR-inhibitory properties. In accordance with this hypothesis, although PKR remained active in DeltaTPR6-expressing cells, the DeltaTPR6 cell line displayed a transformed phenotype and was tumorigenic in nude mice. Thus, the antiapoptotic function of P58(IPK) may be an important factor in its ability to malignantly transform cells. PMID- 10373526 TI - Removal of frameshift intermediates by mismatch repair proteins in Saccharomyces cerevisiae. AB - Frameshift mutations occur when the coding region of a gene is altered by addition or deletion of a number of base pairs that is not a multiple of three. The occurrence of a deletion versus an insertion type of frameshift depends on the nature of the transient intermediate structure formed during DNA synthesis. Extrahelical bases on the template strand give rise to deletions, whereas extrahelical bases on the strand being synthesized produce insertions. We previously used reversion of a +1 frameshift mutation to analyze the role of the mismatch repair (MMR) machinery in correcting -1 frameshift intermediates within a defined region of the yeast LYS2 gene. In this study, we have used reversion of a -1 frameshift mutation within the same region of LYS2 to analyze the role of the MMR machinery in the correction of frameshift intermediates that give rise to insertion events. We found that insertion and deletion events occur at similar rates but that the reversion spectra are very different in both the wild-type and MMR-defective backgrounds. In addition, analysis of the +1 spectra revealed novel roles for Msh3p and Msh6p in removing specific types of frameshift intermediates. PMID- 10373527 TI - Activating phosphorylation of the Kin28p subunit of yeast TFIIH by Cak1p. AB - Cyclin-dependent kinase (CDK)-activating kinases (CAKs) carry out essential activating phosphorylations of CDKs such as Cdc2 and Cdk2. The catalytic subunit of mammalian CAK, MO15/Cdk7, also functions as a subunit of the general transcription factor TFIIH. However, these functions are split in budding yeast, where Kin28p functions as the kinase subunit of TFIIH and Cak1p functions as a CAK. We show that Kin28p, which is itself a CDK, also contains a site of activating phosphorylation on Thr-162. The kinase activity of a T162A mutant of Kin28p is reduced by approximately 75 to 80% compared to that of wild-type Kin28p. Moreover, cells containing kin28(T162A) and a conditional allele of TFB3 (the ortholog of the mammalian MAT1 protein, an assembly factor for MO15 and cyclin H) are severely compromised and display a significant further reduction in Kin28p activity. This finding provides in vivo support for the previous biochemical observation that MO15-cyclin H complexes can be activated either by activating phosphorylation of MO15 or by binding to MAT1. Finally, we show that Kin28p is no longer phosphorylated on Thr-162 following inactivation of Cak1p in vivo, that Cak1p can phosphorylate Kin28p on Thr-162 in vitro, and that this phosphorylation stimulates the CTD kinase activity of Kin28p. Thus, Kin28p joins Cdc28p, the major cell cycle Cdk in budding yeast, as a physiological Cak1p substrate. These findings indicate that although MO15 and Cak1p constitute different forms of CAK, both control the cell cycle and the phosphorylation of the C-terminal domain of the large subunit of RNA polymerase II by TFIIH. PMID- 10373528 TI - Upstream stimulatory factor regulates major histocompatibility complex class I gene expression: the U2DeltaE4 splice variant abrogates E-box activity. AB - The tissue-specific expression of major histocompatibility complex class I genes is determined by a series of upstream regulatory elements, many of which remain ill defined. We now report that a distal E-box element, located between bp -309 and -314 upstream of transcription initiation, acts as a cell type-specific enhancer of class I promoter activity. The class I E box is very active in a neuroblastoma cell line, CHP-126, but is relatively inactive in the HeLa epithelial cell line. The basic helix-loop-helix leucine zipper proteins upstream stimulatory factor 1 (USF1) and USF2 were shown to specifically recognize the class I E box, resulting in the activation of the downstream promoter. Fine mapping of USF1 and USF2 amino-terminal functional domains revealed differences in their abilities to activate the class I E box. Whereas USF1 contained only an extended activation domain, USF2 contained both an activation domain and a negative regulatory region. Surprisingly, the naturally occurring splice variant of USF2 lacking the exon 4 domain, U2DeltaE4, acted as a dominant-negative regulator of USF-mediated activation of the class I promoter. This latter activity is in sharp contrast to the known ability of U2DeltaE4 to activate the adenovirus major late promoter. Class I E-box function is correlated with the relative amount of U2DeltaE4 in a cell, leading to the proposal that U2DeltaE4 modulates class I E-box activity and may represent one mechanism to fine-tune class I expression in various tissues. PMID- 10373529 TI - Activation of phosphatidylinositol 3-kinase in response to interleukin-1 leads to phosphorylation and activation of the NF-kappaB p65/RelA subunit. AB - The work of Reddy et al. (S. A. Reddy, J. A. Huang, and W. S. Liao, J. Biol. Chem. 272:29167-29173, 1997) reveals that phosphatidylinositol 3-kinase (PI3K) plays a role in transducing a signal from the occupied interleukin-1 (IL-1) receptor to nuclear factor kappaB (NF-kappaB), but the underlying mechanism remains to be determined. We have found that IL-1 stimulates interaction of the IL-1 receptor accessory protein with the p85 regulatory subunit of PI3K, leading to the activation of the p110 catalytic subunit. Specific PI3K inhibitors strongly inhibit both PI3K activation and NF-kappaB-dependent gene expression but have no effect on the IL-1-stimulated degradation of IkappaBalpha, the nuclear translocation of NF-kappaB, or the ability of NF-kappaB to bind to DNA. In contrast, PI3K inhibitors block the IL-1-stimulated phosphorylation of NF-kappaB itself, especially the p65/RelA subunit. Furthermore, by using a fusion protein containing the p65/RelA transactivation domain, we found that overexpression of the p110 catalytic subunit of PI3K induces p65/RelA-mediated transactivation and that the specific PI3K inhibitor LY294,002 represses this process. Additionally, the expression of a constitutively activated form of either p110 or the PI3K activated protein kinase Akt also induces p65/RelA-mediated transactivation. Therefore, IL-1 stimulates the PI3K-dependent phosphorylation and transactivation of NF-kappaB, a process quite distinct from the liberation of NF-kappaB from its cytoplasmic inhibitor IkappaB. PMID- 10373530 TI - Induced focal adhesion kinase (FAK) expression in FAK-null cells enhances cell spreading and migration requiring both auto- and activation loop phosphorylation sites and inhibits adhesion-dependent tyrosine phosphorylation of Pyk2. AB - Focal adhesion kinase (FAK) is a nonreceptor protein tyrosine kinase involved in integrin-mediated control of cell behavior. Following cell adhesion to components of the extracellular matrix, FAK becomes phosphorylated at multiple sites, including tyrosines 397, 576, and 577. Tyr-397 is an autophosphorylation site that promotes interaction with c-Src or Fyn. Tyr-576 and Tyr-577 lie in the putative activation loop of the kinase domain, and FAK catalytic activity may be elevated through phosphorylation of these residues by associated Src family kinase. Recent studies have implicated FAK as a positive regulator of cell spreading and migration. To further study the mechanism of adhesion-induced FAK activation and the possible role and signaling requirements for FAK in cell spreading and migration, we utilized the tetracycline repression system to achieve inducible expression of either wild-type FAK or phosphorylation site mutants in fibroblasts derived from FAK-null mouse embryos. Using these Tet-FAK cells, we demonstrated that both the FAK autophosphorylation and activation loop sites are critical for maximum adhesion-induced FAK activation and FAK-enhanced cell spreading and migration responses. Negative effects on cell spreading and migration, as well as decreased phosphorylation of the substrate p130(Cas), were observed upon induced expression of the FAK autophosphorylation site mutant. These negative effects appear to result from an inhibition of integrin-mediated signaling by the FAK-related kinase Pyk2/CAKbeta/RAFTK/CadTK. PMID- 10373531 TI - Tyrosine phosphorylation of the proto-oncoprotein Raf-1 is regulated by Raf-1 itself and the phosphatase Cdc25A. AB - There is a growing body of evidence demonstrating that Raf-1 is phosphorylated on tyrosines upon stimulation of a variety of receptors. Although detection of Raf-1 tyrosine phosphorylation has remained elusive, genetic analyses have demonstrated it to be important for Raf-1 activation. Here we report new findings which indicate that Raf-1 tyrosine phosphorylation is regulated in vivo. In both a mammalian and baculovirus expression system, a kinase-inactive allele of Raf-1 was found to be tyrosine phosphorylated at levels much greater than that of wild type Raf-1. The level of tyrosine phosphate on Raf-1 was markedly increased upon treatment with phosphatase inhibitors either before or after cell lysis. Cdc25A was found to dephosphorylate Raf-1 on tyrosines that resulted in a significant decrease in Raf-1 kinase activity. In NIH 3T3 cells, coexpression of wild-type Raf-1 and phosphatase-inactive Cdc25A led to a marked increase in Raf-1 tyrosine phosphorylation in response to platelet-derived growth factor. These data suggest that the tyrosine phosphorylation of Raf-1 is regulated not only by itself but also by Cdc25A. PMID- 10373532 TI - p53 mediates apoptotic crisis in primary Abelson virus-transformed pre-B cells. AB - Transformation of pre-B cells by Abelson murine leukemia virus (Ab-MLV) involves a balance between positive, growth-stimulatory signals from the v-Abl oncoprotein and negative regulatory cues from cellular genes. This phenomenon is reflected by the clonal selection that occurs during Ab-MLV-mediated transformation in vivo and in vitro. About 50% of all Ab-MLV-transformed pre-B cells express mutant forms of p53 as they emerge from this process, suggesting that this protein may play an important role in the transformation process. Consistent with this idea, expression of p19(Arf), a protein whose function depends on the presence of a functional p53, is required for the apoptotic crisis that characterizes primary Ab-MLV transformants. To test the role of p53 in pre-B-cell transformation directly, we examined the response of Trp53(-/-) mice to Ab-MLV. The absence of p53 shortens the latency of Abelson disease induction but does not affect the frequency of cells susceptible to Ab-MLV-induced transformation. However, primary transformants derived from the null animals bypass the apoptotic crisis that characterizes the transition from primary transformant to fully malignant cell line. These effects do not require p21(Cip-1), a major downstream target of p53; however, consistent with a role of p19(Arf), transformants expressing mutant p53 and abundant p19 retain wild-type p19 sequences. PMID- 10373533 TI - Use of a recombination reporter insert to define meiotic recombination domains on chromosome III of Saccharomyces cerevisiae. AB - In Saccharomyces cerevisiae, meiotic recombination is initiated by DNA double strand breaks (DSBs). DSBs usually occur in intergenic regions that display nuclease hypersensitivity in digests of chromatin. DSBs are distributed nonuniformly across chromosomes; on chromosome III, DSBs are concentrated in two "hot" regions, one in each chromosome arm. DSBs occur rarely in regions within about 40 kb of each telomere and in an 80-kb region in the center of the chromosome, just to the right of the centromere. We used recombination reporter inserts containing arg4 mutant alleles to show that the "cold" properties of the central DSB-deficient region are imposed on DNA inserted in the region. Cold region inserts display DSB and recombination frequencies that are substantially less than those seen with similar inserts in flanking hot regions. This occurs without apparent change in chromatin structure, as the same pattern and level of DNase I hypersensitivity is seen in chromatin of hot and cold region inserts. These data are consistent with the suggestion that features of higher-order chromosome structure or chromosome dynamics act in a target sequence-independent manner to control where recombination events initiate during meiosis. PMID- 10373534 TI - BRCA1 is phosphorylated at serine 1497 in vivo at a cyclin-dependent kinase 2 phosphorylation site. AB - BRCA1 is a cell cycle-regulated nuclear protein that is phosphorylated mainly on serine and to a lesser extent on threonine residues. Changes in phosphorylation occur in response to cell cycle progression and DNA damage. Specifically, BRCA1 undergoes hyperphosphorylation during late G1 and S phases of the cell cycle. Here we report that BRCA1 is phosphorylated in vivo at serine 1497 (S1497), which is part of a cyclin-dependent kinase (CDK) consensus site. S1497 can be phosphorylated in vitro by CDK2-cyclin A or E. BRCA1 coimmunoprecipitates with an endogenous serine-threonine protein kinase activity that phosphorylates S1497 in vitro. This cellular kinase activity is sensitive to transfection of a dominant negative form of CDK2 as well as the application of the CDK inhibitors p21 and butyrolactone I but not p16. Furthermore, BRCA1 coimmunoprecipitates with CDK2 and cyclin A. These results suggest that the endogenous kinase activity is composed of CDK2-cyclin complexes, at least in part, concordant with the G1/S specific increase in BRCA1 phosphorylation. PMID- 10373535 TI - Repressive effect of Sp1 on the C/EBPalpha gene promoter: role in adipocyte differentiation. AB - Expression of C/EBPalpha is required for differentiation of 3T3-L1 preadipocytes into adipocytes. Previous investigations indicated that transcription of the C/EBPalpha gene is sequentially activated during differentiation, initially by C/EBPbeta and C/EBPdelta and later by C/EBPalpha (autoactivation). These events are mediated by a C/EBP regulatory element in the promoter of the C/EBPalpha gene. This article presents evidence that members of the Sp family, notably Sp1, act repressively on the C/EBPalpha promoter prior to the induction of differentiation. Sp1 was shown to bind to a GC box at the 5' end of the C/EBP regulatory element in the C/EBPalpha promoter and, in so doing, to competitively prevent binding to and transactivation of the promoter by the C/EBPs. One of the differentiation inducers methylisobutylxanthine (a cAMP phosphodiesterase inhibitor) or Forskolin, both of which increase the cellular cAMP level, causes down-regulation of Sp1. This decrease in Sp1 level early in the differentiation program appears to facilitate access of C/EBPbeta and/or C/EBPdelta to the C/EBP regulatory element and, thereby, derepression of the C/EBPalpha gene. PMID- 10373536 TI - Fanconi anemia proteins FANCA, FANCC, and FANCG/XRCC9 interact in a functional nuclear complex. AB - Fanconi anemia (FA) is an autosomal recessive cancer susceptibility syndrome with at least eight complementation groups (A to H). Three FA genes, corresponding to complementation groups A, C, and G, have been cloned, but their cellular function remains unknown. We have previously demonstrated that the FANCA and FANCC proteins interact and form a nuclear complex in normal cells, suggesting that the proteins cooperate in a nuclear function. In this report, we demonstrate that the recently cloned FANCG/XRCC9 protein is required for binding of the FANCA and FANCC proteins. Moreover, the FANCG protein is a component of a nuclear protein complex containing FANCA and FANCC. The amino-terminal region of the FANCA protein is required for FANCG binding, FANCC binding, nuclear localization, and functional activity of the complex. Our results demonstrate that the three cloned FA proteins cooperate in a large multisubunit complex. Disruption of this complex results in the specific cellular and clinical phenotype common to most FA complementation groups. PMID- 10373537 TI - Cyclic AMP-dependent protein kinase regulates pseudohyphal differentiation in Saccharomyces cerevisiae. AB - In response to nitrogen starvation, diploid cells of the yeast Saccharomyces cerevisiae differentiate to a filamentous growth form known as pseudohyphal differentiation. Filamentous growth is regulated by elements of the pheromone mitogen-activated protein (MAP) kinase cascade and a second signaling cascade involving the receptor Gpr1, the Galpha protein Gpa2, Ras2, and cyclic AMP (cAMP). We show here that the Gpr1-Gpa2-cAMP pathway signals via the cAMP dependent protein kinase, protein kinase A (PKA), to regulate pseudohyphal differentiation. Activation of PKA by mutation of the regulatory subunit Bcy1 enhances filamentous growth. Mutation and overexpression of the PKA catalytic subunits reveal that the Tpk2 catalytic subunit activates filamentous growth, whereas the Tpk1 and Tpk3 catalytic subunits inhibit filamentous growth. The PKA pathway regulates unipolar budding and agar invasion, whereas the MAP kinase cascade regulates cell elongation and invasion. Epistasis analysis supports a model in which PKA functions downstream of the Gpr1 receptor and the Gpa2 and Ras2 G proteins. Activation of filamentous growth by PKA does not require the transcription factors Ste12 and Tec1 of the MAP kinase cascade, Phd1, or the PKA targets Msn2 and Msn4. PKA signals pseudohyphal growth, in part, by regulating Flo8-dependent expression of the cell surface flocculin Flo11. In summary, the cAMP-dependent protein kinase plays an intimate positive and negative role in regulating filamentous growth, and these findings may provide insight into the roles of PKA in mating, morphogenesis, and virulence in other yeasts and pathogenic fungi. PMID- 10373538 TI - Cell cycle control of Cdc7p kinase activity through regulation of Dbf4p stability. AB - In Saccharomyces cerevisiae, the heteromeric kinase complex Cdc7p-Dbf4p plays a pivotal role at replication origins in triggering the initiation of DNA replication during the S phase. We have assayed the kinase activity of endogenous levels of Cdc7p kinase by using a likely physiological target, Mcm2p, as a substrate. Using this assay, we have confirmed that Cdc7p kinase activity fluctuates during the cell cycle; it is low in the G1 phase, rises as cells enter the S phase, and remains high until cells complete mitosis. These changes in kinase activity cannot be accounted for by changes in the levels of the catalytic subunit Cdc7p, as these levels are constant during the cell cycle. However, the fluctuations in kinase activity do correlate with levels of the regulatory subunit Dbf4p. The regulation of Dbf4p levels can be attributed in part to increased degradation of the protein in G1 cells. This G1-phase instability is cdc16 dependent, suggesting a role of the anaphase-promoting complex in the turnover of Dbf4p. Overexpression of Dbf4p in the G1 phase can partially overcome this elevated turnover and lead to an increase in Cdc7p kinase activity. Thus, the regulation of Dbf4p levels through the control of Dbf4p degradation has an important role in the regulation of Cdc7p kinase activity during the cell cycle. PMID- 10373539 TI - Activation of a delayed-early gene encoding MHR3 by the ecdysone receptor heterodimer EcR-B1-USP-1 but not by EcR-B1-USP-2. AB - MHR3, a homolog of the retinoid orphan receptor (ROR), is a transcription factor in the nuclear hormone receptor family that is induced by 20-hydroxyecdysone (20E) in the epidermis of the tobacco hornworm, Manduca sexta. Its 2.7-kb 5' flanking region was found to contain four putative ecdysone receptor response elements (EcREs) and a monomeric (GGGTCA) nuclear receptor binding site. Activation of this promoter fused to a chloramphenicol acetyltransferase (CAT) reporter by 2 micrograms of 20E per ml in Manduca GV1 cells was similar to that of endogenous MHR3, with detectable CAT by 3 h. When the ecdysone receptor B1 (EcR-B1) and Ultraspiracle 1 (USP-1) were expressed at high levels under the control of a constitutive promoter, CAT levels after a 3-h exposure to 20E increased two- to sixfold. In contrast, high expression of EcR-B1 and USP-2 caused little increase in CAT levels in response to 20E. Moreover, expression of USP-2 prevented activation by EcR-B1-USP-1. Deletion experiments showed that the upstream region, including the three most proximal putative EcREs, was responsible for most of the 20E activation, with the EcRE3 at -671 and the adjacent GGGTCA being most critical. The EcRE1 at -342 was necessary but not sufficient for the activational response but was the only one of the three putative EcREs to bind the EcR-B1-USP-1 complex in gel mobility shift assays and was responsible for the silencing action of EcR-B1-USP-1 in the absence of hormone. EcRE2 and EcRE3 each specifically bound other protein(s) in the cell extract, but not EcR and USP, and so are not EcREs in this cellular context. When cell extracts were used, the EcR-B1-USP-2 heterodimer showed no binding to EcRE1, and the presence of excess USP-2 prevented the binding of EcR-B1-USP-1 to this element. In contrast, in vitro-transcribed-translated USP-1 and USP-2 both formed heterodimeric complexes with EcR-B1 that bound ponasterone A with the same Kd (7 x 10(-10) M) and bound to both EcRE1 and heat shock protein 27 EcRE. Thus, factors present in the cell extract appear to modulate the differential actions of the two USP isoforms. PMID- 10373540 TI - Enhancer-dependent transcriptional oscillations in mouse erythroleukemia cells. AB - By using recombinase-mediated cassette exchange, a method that allows integration of single copies of different constructs at the same predetermined chromosomal location, several expression cassettes have been integrated at a randomly chosen locus in the genome of mouse erythroleukemia cells. The cassettes studied contain the human beta-globin promoter fused to lacZ coding sequences either alone or linked to DNase I-hypersensitive site HS2, HS3, or HS234 (a large locus control region fragment containing HS2, HS3, and HS4) of the human beta-globin locus control region. Analysis of expression of these cassettes revealed mosaic expression patterns reminiscent of, but clearly different from, position effect variegation. Further investigations demonstrated that these mosaic expression patterns are caused by dynamic activation and inactivation of the transcription unit, resulting in oscillations of expression. These oscillations occur once in every few cell cycles at a rate specific for the enhancer present at the locus. DNase I sensitivity studies revealed that the chromatin is accessible and that DNase-hypersensitive sites were present whether or not the transcription unit is active, suggesting that the oscillations occur between transcriptionally competent and transcriptionally active chromatin conformations, rather than between open and closed chromatin conformations. Treatment of oscillating cells with trichostatin A eliminates the oscillations only after the cells have passed through late G1 or early S, suggesting that these oscillations might be caused by changes in histone acetylation patterns. PMID- 10373541 TI - Homeoproteins CDP and SATB1 interact: potential for tissue-specific regulation. AB - Homeoproteins are known to participate in development and cell type specification. The homeoproteins CCAAT displacement protein (CDP) and special AT rich sequence binding protein 1 (SATB1) have been shown to bind to nuclear matrix associated regions and to act as repressors of many cellular genes. Moreover, binding of SATB1 to the mouse mammary tumor virus (MMTV) promoter region dramatically affects the tissue-specific transcription of this retrovirus. Because protein-protein interactions are a common means of regulating homeoprotein function, we tested whether SATB1 and CDP interact in vivo and in vitro. SATB1 interacted with CDP through its DNA-binding domain, as demonstrated by glutathione S-transferase (GST) pull-down assays. GST pull-down assays also showed that CDP associated with SATB1 through three of its four DNA-binding domains (CR1, CR2, and the homeodomain). SATB1-specific antisera, but not preimmune sera, precipitated CDP from nuclear extracts, and CDP-specific antisera precipitated SATB1 from the same extracts. Far-Western blotting detected interaction of SATB1 and CDP in several different tissue extracts. Association of purified SATB1 and CDP in vitro resulted in the inability of each protein to bind to DNA in gel retardation assays. CDP overexpression in cultured T cells led to a loss of detectable SATB1 binding to the MMTV promoter region, as measured by gel shift experiments. CDP overexpression also elevated MMTV long terminal repeat reporter gene activity in transient-transfection assays, a result consistent with neutralization of the SATB1 repressor function in T cells. SATB1 is very abundant in certain tissues, particularly thymus, whereas CDP is relatively ubiquitous, except in certain terminally differentiated cell types. Because of the tissue and cell type distribution of SATB1 and CDP, we propose that the SATB1-to-CDP ratio in different tissues is a novel mechanism for homeoproteins to control gene expression and differentiation in mammals. PMID- 10373542 TI - Activation of RNA polymerase III transcription in cells transformed by simian virus 40. AB - RNA polymerase (Pol) III transcription is abnormally active in fibroblasts that have been transformed by simian virus 40 (SV40). This report presents evidence that two separate components of the general Pol III transcription apparatus, TFIIIB and TFIIIC2, are deregulated following SV40 transformation. TFIIIC2 subunits are expressed at abnormally high levels in SV40-transformed cells, an effect which is observed at both protein and mRNA levels. In untransformed fibroblasts, TFIIIB is subject to repression through association with the retinoblastoma protein RB. The interaction between RB and TFIIIB is compromised following SV40 transformation. Furthermore, the large T antigen of SV40 is shown to relieve repression by RB. The E7 oncoprotein of human papillomavirus can also activate Pol III transcription, an effect that is dependent on its ability to bind to RB. The data provide evidence that both TFIIIB and TFIIIC2 are targets for activation by DNA tumor viruses. PMID- 10373543 TI - The naturally occurring mutants of DDB are impaired in stimulating nuclear import of the p125 subunit and E2F1-activated transcription. AB - The human UV-damaged-DNA binding protein DDB has been linked to the repair deficiency disease xeroderma pigmentosum group E (XP-E), because a subset of XP-E patients lack the damaged-DNA binding function of DDB. Moreover, the microinjection of purified DDB complements the repair deficiency in XP-E cells lacking DDB. Two naturally occurring XP-E mutations of DDB, 82TO and 2RO, have been characterized. They have single amino acid substitutions (K244E and R273H) within the WD motif of the p48 subunit of DDB, and the mutated proteins lack the damaged-DNA binding activity. In this report, we describe a new function of the p48 subunit of DDB, which reveals additional defects in the function of the XP-E mutants. We show that when the subunits of DDB were expressed individually, p48 localized in the nucleus and p125 localized in the cytoplasm. The coexpression of p125 with p48 resulted in an increased accumulation of p125 in the nucleus, indicating that p48 plays a critical role in the nuclear localization of p125. The mutant forms of p48, 2RO and 82TO, are deficient in stimulating the nuclear accumulation of the p125 subunit of DDB. In addition, the mutant 2RO fails to form a stable complex with the p125 subunit of DDB. Our previous studies indicated that DDB can associate with the transcription factor E2F1 and can function as a transcriptional partner of E2F1. Here we show that the two mutants, while they associate with E2F1 as efficiently as wild-type p48, are severely impaired in stimulating E2F1-activated transcription. This is consistent with our observation that both subunits of DDB are required to stimulate E2F1-activated transcription. The results provide insights into the functions of the subunits of DDB and suggest a possible link between the role of DDB in E2F1-activated transcription and the repair deficiency disease XP-E. PMID- 10373545 TI - Transcriptional activity and chromatin structure of enhancer-deleted rRNA genes in Saccharomyces cerevisiae. AB - We used the psoralen gel retardation assay and Northern blot analysis in an in vivo yeast system to analyze effects of rDNA enhancer deletions on the chromatin structure and the transcription of tagged rDNA units. We found that upon deletion of a single enhancer element, transcription of the upstream and downstream rRNA gene was reduced by about 50%. Although removing both flanking enhancers of an rRNA gene led to a further reduction in transcription levels, a significant amount of transcriptional activity remained, either resulting from the influence of more distantly located enhancer elements or reflecting the basal activity of the polymerase I promoter within the nucleolus. Despite the reduction of transcriptional activity upon enhancer deletion, the activation frequency (proportion of nonnucleosomal to nucleosomal gene copies in a given cell culture) of the tagged rRNA genes was not significantly altered, as determined by the psoralen gel retardation assay. This is a strong indication that, within the nucleolus, the yeast rDNA enhancer functions by increasing transcription rates of active rRNA genes and not by activating silent transcription units. PMID- 10373544 TI - Cloning and characterization of two evolutionarily conserved subunits (TFIIIC102 and TFIIIC63) of human TFIIIC and their involvement in functional interactions with TFIIIB and RNA polymerase III. AB - Human transcription factor IIIC (hTFIIIC) is a multisubunit complex that mediates transcription of class III genes through direct recognition of promoters (for tRNA and virus-associated RNA genes) or promoter-TFIIIA complexes (for the 5S RNA gene) and subsequent recruitment of TFIIIB and RNA polymerase III. We describe the cognate cDNA cloning and characterization of two subunits (hTFIIIC63 and hTFIIIC102) that are present within a DNA-binding subcomplex (TFIIIC2) of TFIIIC and are related in structure and function to two yeast TFIIIC subunits (yTFIIIC95 and yTFIIIC131) previously shown to interact, respectively, with the promoter (A box) and with a subunit of yeast TFIIIB. hTFIIIC63 and hTFIIIC102 show parallel in vitro interactions with the homologous human TFIIIB and RNA polymerase III components, as well as additional interactions that may facilitate both TFIIIB and RNA polymerase III recruitment. These include novel interactions of hTFIIIC63 with hTFIIIC102, with hTFIIIB90, and with hRPC62, in addition to the hTFIIIC102 hTFIIIB90 and hTFIIIB90-hRPC39 interactions that parallel the previously described interactions in yeast. As reported for yTFIIIC131, hTFIIIC102 contains acidic and basic regions, tetratricopeptide repeats (TPRs), and a helix-loop helix domain, and mutagenesis studies have implicated the TPRs in interactions both with hTFIIIC63 and with hTFIIIB90. These observations further document conservation from yeast to human of the structure and function of the RNA polymerase III transcription machinery, but in addition, they provide new insights into the function of hTFIIIC and suggest direct involvement in recruitment of both TFIIIB and RNA polymerase III. PMID- 10373546 TI - Physiological requirement for both SH2 domains for phospholipase C-gamma1 function and interaction with platelet-derived growth factor receptors. AB - Two approaches have been utilized to investigate the role of individual SH2 domains in growth factor activation of phospholipase C-gamma1 (PLC-gamma1). Surface plasmon resonance analysis indicates that the individual N-SH2 and C-SH2 domains are able to specifically recognize a phosphotyrosine-containing peptide corresponding to Tyr 1021 of the platelet-derived growth factor (PDGF) beta receptor. To assess SH2 function in the context of the full-length PLC-gamma1 molecule as well as within the intact cell, PLC-gamma1 SH2 domain mutants, disabled by site-directed mutagenesis of the N-SH2 and/or C-SH2 domain(s), were expressed in Plcg1(-/-) fibroblasts. Under equilibrium incubation conditions (4 degrees C, 40 min), the N-SH2 domain, but not the C-SH2 domain, was sufficient to mediate significant PLC-gamma1 association with the activated PDGF receptor and PLC-gamma1 tyrosine phosphorylation. When both SH2 domains in PLC-gamma1 were disabled, the double mutant did not associate with activated PDGF receptors and was not tyrosine phosphorylated. However, no single SH2 mutant was able to mediate growth factor activation of Ca2+ mobilization or inositol 1,4,5 trisphosphate (IP3) formation. Subsequent kinetic experiments demonstrated that each single SH2 domain mutant was significantly impaired in its capacity to mediate rapid association with activated PDGF receptors and become tyrosine phosphorylated. Hence, when assayed under physiological conditions necessary to achieve a rapid biological response (Ca2+ mobilization and IP3 formation), both SH2 domains of PLC-gamma1 are essential to growth factor responsiveness. PMID- 10373547 TI - Utilization of splicing elements and polyadenylation signal elements in the coupling of polyadenylation and last-intron removal. AB - Polyadenylation (PA) is the process by which the 3' ends of most mammalian mRNAs are formed. In nature, PA is highly coordinated, or coupled, with splicing. In mammalian systems, the most compelling mechanistic model for coupling arises from data supporting exon definition (2, 34, 37). We have examined the roles of individual functional components of splicing and PA signals in the coupling process by using an in vitro splicing and PA reaction with a synthetic pre-mRNA substrate containing an adenovirus splicing cassette and the simian virus 40 late PA signal. The effects of individually mutating splicing elements and PA elements in this substrate were determined. We found that mutation of the polypyrimidine tract and the 3' splice site significantly reduced PA efficiency and that mutation of the AAUAAA and the downstream elements of the PA signal decreased splicing efficiency, suggesting that these elements are the most significant for the coupling of splicing and PA. Although mutation of the upstream elements (USEs) of the PA signal dramatically decreased PA, splicing was only modestly affected, suggesting that USEs modestly affect coupling. Mutation of the 5' splice site in the presence of a viable polypyrimidine tract and the 3' splice site had no effect on PA, suggesting no effect of this element on coupling. However, our data also suggest that a site for U1 snRNP binding (e.g., a 5' splice site) within the last exon can negatively effect both PA and splicing; hence, a 5' splice site-like sequence in this position appears to be a modulator of coupling. In addition, we show that the RNA-protein complex formed to define an exon may inhibit processing if the definition of an adjacent exon fails. This finding indicates a mechanism for monitoring the appropriate definition of exons and for allowing only pre-mRNAs with successfully defined exons to be processed. PMID- 10373548 TI - SOCS-3 is tyrosine phosphorylated in response to interleukin-2 and suppresses STAT5 phosphorylation and lymphocyte proliferation. AB - Members of the recently discovered SOCS/CIS/SSI family have been proposed as regulators of cytokine signaling, and while targets and mechanisms have been suggested for some family members, the precise role of these proteins remains to be defined. To date no SOCS proteins have been specifically implicated in interleukin-2 (IL-2) signaling in T cells. Here we report SOCS-3 expression in response to IL-2 in both T-cell lines and human peripheral blood lymphocytes. SOCS-3 protein was detectable as early as 30 min following IL-2 stimulation, while CIS was seen only at low levels after 2 h. Unlike CIS, SOCS-3 was rapidly tyrosine phosphorylated in response to IL-2. Tyrosine phosphorylation of SOCS-3 was observed upon coexpression with Jak1 and Jak2 but only weakly with Jak3. In these experiments, SOCS-3 associated with Jak1 and inhibited Jak1 phosphorylation, and this inhibition was markedly enhanced by the presence of IL 2 receptor beta chain (IL-2Rbeta). Moreover, following IL-2 stimulation of T cells, SOCS-3 was able to interact with the IL-2 receptor complex, and in particular tyrosine phosphorylated Jak1 and IL-2Rbeta. Additionally, in lymphocytes expressing SOCS-3 but not CIS, IL-2-induced tyrosine phosphorylation of STAT5b was markedly reduced, while there was only a weak effect on IL-3 mediated STAT5b tyrosine phosphorylation. Finally, proliferation induced by both IL-2- and IL-3 was significantly inhibited in the presence of SOCS-3. The findings suggest that when SOCS-3 is rapidly induced by IL-2 in T cells, it acts to inhibit IL-2 responses in a classical negative feedback loop. PMID- 10373549 TI - Mechanism of protein kinase B activation by cyclic AMP-dependent protein kinase. AB - Activation of protein kinase B (PKB) by growth factors and hormones has been demonstrated to proceed via phosphatidylinositol 3-kinase (PI3-kinase). In this report, we show that PKB can also be activated by PKA (cyclic AMP [cAMP] dependent protein kinase) through a PI3-kinase-independent pathway. Although this activation required phosphorylation of PKB, PKB is not likely to be a physiological substrate of PKA since a mutation in the sole PKA consensus phosphorylation site of PKB did not abolish PKA-induced activation of PKB. In addition, mechanistically, this activation was different from that of growth factors since it did not require phosphorylation of the S473 residue, which is essential for full PKB activation induced by insulin. These data were supported by the fact that mutation of residue S473 of PKB to alanine did not prevent it from being activated by forskolin. Moreover, phosphopeptide maps of overexpressed PKB from COS cells showed differences between insulin- and forskolin-stimulated cells that pointed to distinct activation mechanisms of PKB depending on whether insulin or cAMP was used. We looked at events downstream of PKB and found that PKA activation of PKB led to the phosphorylation and inhibition of glycogen synthase kinase-3 (GSK-3) activity, a known in vivo substrate of PKB. Overexpression of a dominant negative PKB led to the loss of inhibition of GSK-3 in both insulin- and forskolin-treated cells, demonstrating that PKB was responsible for this inhibition in both cases. Finally, we show by confocal microscopy that forskolin, similar to insulin, was able to induce translocation of PKB to the plasma membrane. This process was inhibited by high concentrations of wortmannin (300 nM), suggesting that forskolin-induced PKB movement may require phospholipids, which are probably not generated by class I or class III PI3-kinase. However, high concentrations of wortmannin did not abolish PKB activation, which demonstrates that translocation per se is not important for PKA induced PKB activation. PMID- 10373550 TI - Human Cdc34 and Rad6B ubiquitin-conjugating enzymes target repressors of cyclic AMP-induced transcription for proteolysis. AB - Ubiquitin-mediated proteolysis controls diverse physiological processes in eukaryotes. However, few in vivo targets of the mammalian Cdc34 and Rad6 ubiquitin-conjugating enzymes are known. A yeast-based genetic assay to identify proteins that interact with human Cdc34 resulted in three cDNAs encoding bZIP DNA binding motifs. Two of these interactants are repressors of cyclic AMP (cAMP) induced transcription: hICERIIgamma, a product of the CREM gene, and hATF5, a novel ATF homolog. Transfection assays with mammalian cells demonstrate both hCdc34- and hRad6B-dependent ubiquitin-mediated proteolysis of hICERIIgamma and hATF5. This degradation requires an active ubiquitin-conjugating enzyme and results in abrogation of ICERIIgamma- and ATF5-mediated repression of cAMP induced transcription. Consistent with these results, the endogenous ICER protein is elevated in cells which are null for murine Rad6B (mHR6B-/-) or transfected with dominant negative and antisense constructs of human CDC34. Based on the requirement for CREM/ICER and Rad6B proteins in spermatogenesis, we determined expression of Cdc34, Rad6B, CREM/ICER isoforms, and the Skp1-Cullin-F-box ubiquitin protein ligase subunits Cul-1 and Cul-2, which are associated with Cdc34 activity during murine testicular development. Cdc34, Rad6B, and the Cullin proteins are expressed in a developmentally regulated manner, with distinctly different patterns for Cdc34 and the Cullin proteins in germ cells. The Cdc34 and Rad6B proteins are significantly elevated in meiotic and postmeiotic haploid germ cells when chromatin modifications occur. Thus, the stability of specific mammalian transcription factors is the result of complex targeting by multiple ubiquitin-conjugating enzymes and may have an impact on cAMP-inducible gene regulation during both meiotic and mitotic cell cycles. PMID- 10373551 TI - Regulation of nuclear localization and transcriptional activity of TFII-I by Bruton's tyrosine kinase. AB - Bruton's tyrosine kinase (Btk) is required for normal B-cell development, as defects in Btk lead to X-linked immunodeficiency (xid) in mice and X-linked agammaglobulinemia (XLA) in humans. Here we demonstrate a functional interaction between the multifunctional transcription factor TFII-I and Btk. Ectopic expression of wild-type Btk enhances TFII-I-mediated transcriptional activation and its tyrosine phosphorylation in transient-transfection assays. Mutation of Btk in either the PH domain (R28C, as in the murine xid mutation) or the kinase domain (K430E) compromises its ability to enhance both the tyrosine phosphorylation and the transcriptional activity of TFII-I. TFII-I associates constitutively in vivo with wild-type Btk and kinase-inactive Btk but not xid Btk. However, membrane immunoglobulin M cross-linking in B cells leads to dissociation of TFII-I from Btk. We further show that while TFII-I is found in both the nucleus and cytoplasm of wild-type and xid primary resting B cells, nuclear TFII-I is greater in xid B cells. Most strikingly, receptor cross-linking of wild-type (but not xid) B cells results in increased nuclear import of TFII-I. Taken together, these data suggest that although the PH domain of Btk is primarily responsible for its physical interaction with TFII-I, an intact kinase domain of Btk is required to enhance transcriptional activity of TFII-I in the nucleus. Thus, mutations impairing the physical and/or functional association between TFII-I and Btk may result in diminished TFII-I-dependent transcription and contribute to defective B-cell development and/or function. PMID- 10373552 TI - Disordered T-cell development and T-cell malignancies in SCL LMO1 double transgenic mice: parallels with E2A-deficient mice. AB - The gene most commonly activated by chromosomal rearrangements in patients with T cell acute lymphoblastic leukemia (T-ALL) is SCL/tal. In collaboration with LMO1 or LMO2, the thymic expression of SCL/tal leads to T-ALL at a young age with a high degree of penetrance in transgenic mice. We now show that SCL LMO1 double transgenic mice display thymocyte developmental abnormalities in terms of proliferation, apoptosis, clonality, and immunophenotype prior to the onset of a frank malignancy. At 4 weeks of age, thymocytes from SCL LMO1 mice show 70% fewer total thymocytes, with increased rates of both proliferation and apoptosis, than control thymocytes. At this age, a clonal population of thymocytes begins to populate the thymus, as evidenced by oligoclonal T-cell-receptor gene rearrangements. Also, there is a dramatic increase in immature CD44(+) CD25(-) cells, a decrease in the more mature CD4(+) CD8(+) cells, and development of an abnormal CD44(+) CD8(+) population. An identical pattern of premalignant changes is seen with either a full-length SCL protein or an amino-terminal truncated protein which lacks the SCL transactivation domain, demonstrating that the amino terminal portion of SCL is not important for leukemogenesis. Lastly, we show that the T-ALL which develop in the SCL LMO1 mice are strikingly similar to those which develop in E2A null mice, supporting the hypothesis that SCL exerts its oncogenic action through a functional inactivation of E proteins. PMID- 10373553 TI - Distinct glucocorticoid receptor transcriptional regulatory surfaces mediate the cytotoxic and cytostatic effects of glucocorticoids. AB - Glucocorticoids act through the glucocorticoid receptor (GR), which can function as a transcriptional activator or repressor, to elicit cytostatic and cytotoxic effects in a variety of cells. The molecular mechanisms regulating these events and the target genes affected by the activated receptor remain largely undefined. Using cultured human osteosarcoma cells as a model for the GR antiproliferative effect, we demonstrate that in U20S cells, GR activation leads to irreversible growth inhibition, apoptosis, and repression of Bcl2. This cytotoxic effect is mediated by GR's transcriptional repression function, since transactivation deficient mutants and ligands still bring about apoptosis and Bcl2 down regulation. In contrast, the antiproliferative effect of GR in SAOS2 cells is reversible, does not result in apoptosis or repression of Bcl2, and is a function of the receptor's ability to stimulate transcription. Thus, the cytotoxic versus cytostatic outcome of glucocorticoid treatment is cell context dependent. Interestingly, the cytostatic effect of glucocorticoids in SAOS2 cells involves multiple GR activation surfaces. GR mutants and ligands that disrupt individual transcriptional activation functions (activation function 1 [AF-1] and AF-2) or receptor dimerization fail to fully inhibit cellular proliferation and, remarkably, discriminate between the targets of GR's cytostatic action, the cyclin-dependent kinase inhibitors p21(Cip1) and p27(Kip1). Induction of p21(Cip1) is agonist dependent and requires AF-2 but not AF-1 or GR dimerization. In contrast, induction of p27(Kip1) is agonist independent, does not require AF-2 or AF-1, but depends on GR dimerization. Our findings indicate that multiple GR transcriptional regulatory mechanisms that employ distinct receptor surfaces are used to evoke either the cytostatic or cytotoxic response to glucocorticoids. PMID- 10373554 TI - Synergistic transcriptional activation by TATA-binding protein and hTAFII28 requires specific amino acids of the hTAFII28 histone fold. AB - Coexpression of the human TATA-binding protein (TBP)-associated factor 28 (hTAFII28) with the altered-specificity mutant TBP spm3 synergistically enhances transcriptional activation by the activation function 2 of the nuclear receptors (NRs) for estrogen and vitamin D3 from a reporter plasmid containing a TGTA element in mammalian cells. This synergy is abolished by mutation of specific amino acids in the alpha2-helix of the histone fold in the conserved C-terminal region of hTAFII28. Critical amino acids are found on both the exposed hydrophilic face of this helix and the hydrophobic interface with TAFII18. This alpha-helix of hTAFII28 therefore mediates multiple interactions required for coactivator activity. We further show that mutation of specific residues in the H1' alpha-helix of TBP either reduces or increases interactions with hTAFII28. The mutations which reduce interactions with hTAFII28 do not affect functional synergy, whereas the TBP mutation which increases interaction with hTAFII28 is defective in its ability to synergistically enhance activation by NRs. However, this TBP mutant supports activation by other activators and is thus specifically defective for its ability to synergize with hTAFII28. PMID- 10373555 TI - Domain swapping used to investigate the mechanism of protein kinase B regulation by 3-phosphoinositide-dependent protein kinase 1 and Ser473 kinase. AB - Protein kinase B (PKB or Akt), a downstream effector of phosphoinositide 3-kinase (PI 3-kinase), has been implicated in insulin signaling and cell survival. PKB is regulated by phosphorylation on Thr308 by 3-phosphoinositide-dependent protein kinase 1 (PDK1) and on Ser473 by an unidentified kinase. We have used chimeric molecules of PKB to define different steps in the activation mechanism. A chimera which allows inducible membrane translocation by lipid second messengers that activate in vivo protein kinase C and not PKB was created. Following membrane attachment, the PKB fusion protein was rapidly activated and phosphorylated at the two key regulatory sites, Ser473 and Thr308, in the absence of further cell stimulation. This finding indicated that both PDK1 and the Ser473 kinase may be localized at the membrane of unstimulated cells, which was confirmed for PDK1 by immunofluorescence studies. Significantly, PI 3-kinase inhibitors prevent the phosphorylation of both regulatory sites of the membrane-targeted PKB chimera. Furthermore, we show that PKB activated at the membrane was rapidly dephosphorylated following inhibition of PI 3-kinase, with Ser473 being a better substrate for protein phosphatase. Overall, the results demonstrate that PKB is stringently regulated by signaling pathways that control both phosphorylation/activation and dephosphorylation/inactivation of this pivotal protein kinase. PMID- 10373556 TI - Cell cycle withdrawal promotes myogenic induction of Akt, a positive modulator of myocyte survival. AB - During myogenesis, proliferating myoblasts withdraw from the cell cycle, acquire an apoptosis-resistant phenotype, and differentiate into myotubes. Previous studies indicate that myogenic induction of the cyclin-dependent kinase inhibitor p21 results in an inhibition of apoptotic cell death in addition to its role as a negative cell cycle regulator. Here we demonstrate that the protein encoded by the Akt proto-oncogene is induced in C2C12 cells during myogenic differentiation with a corresponding increase in kinase activity. In differentiating cultures, expression of dominant-negative forms of Akt increase the frequency of cell death whereas expression of wild-type Akt protects against death, indicating that Akt is a positive modulator of myocyte survival. Antisense oligonucleotides against p21 block cell cycle withdrawal, inhibit Akt induction, and enhance cell death in differentiating myocyte cultures. Adenovirus-mediated transfer of wild-type or constitutively active Akt constructs confer partial resistance to cell death under conditions where cell cycle exit is blocked by the antisense oligonucleotides. Collectively, these data indicate that cell cycle withdrawal facilitates the induction of Akt during myogenesis, promoting myocyte survival. PMID- 10373557 TI - A novel growth- and cell cycle-regulated protein, ASK, activates human Cdc7 related kinase and is essential for G1/S transition in mammalian cells. AB - A novel human protein, ASK (activator of S phase kinase), was identified on the basis of its ability to bind to human Cdc7-related kinase (huCdc7). ASK forms an active kinase complex with huCdc7 that is capable of phosphorylating MCM2 protein. ASK appears to be the major activator of huCdc7, since immunodepletion of ASK protein from the extract is accompanied by the loss of huCdc7-dependent kinase activity. Expression of ASK is regulated by growth factor stimulation, and levels oscillate through the cell cycle, reaching a peak during S phase. Concomitantly, the huCdc7-dependent kinase activity significantly increases when cells are in S phase. Furthermore, we have demonstrated that ASK serves an essential function for entry into S phase by showing that microinjection of ASK specific antibodies into mammalian cells inhibited DNA replication. Our data show that ASK is a novel cyclin-like regulatory subunit of the huCdc7 kinase complex and that it plays a pivotal role in G1/S transition in mammalian cells. PMID- 10373558 TI - Atm inactivation results in aberrant telomere clustering during meiotic prophase. AB - A-T (ataxia telangiectasia) individuals frequently display gonadal atrophy, and Atm-/- mice show spermatogenic failure due to arrest at prophase of meiosis I. Chromosomal movements take place during meiotic prophase, with telomeres congregating on the nuclear envelope to transiently form a cluster during the leptotene/zygotene transition (bouquet arrangement). Since the ATM protein has been implicated in telomere metabolism of somatic cells, we have set out to investigate the effects of Atm inactivation on meiotic telomere behavior. Fluorescent in situ hybridization and synaptonemal complex (SC) immunostaining of structurally preserved spermatocytes I revealed that telomere clustering occurs aberrantly in Atm-/- mice. Numerous spermatocytes of Atm-/- mice displayed locally accumulated telomeres with stretches of SC near the clustered chromosome ends. This contrasted with spermatogenesis of normal mice, where only a few leptotene/zygotene spermatocytes I with clustered telomeres were detected. Pachytene nuclei, which were much more abundant in normal mice, displayed telomeres scattered over the nuclear periphery. It appears that the timing and occurrence of chromosome polarization is altered in Atm-/- mice. When we examined telomere-nuclear matrix interactions in spermatocytes I, a significant difference was observed in the ratio of soluble versus matrix-associated telomeric DNA sequences between meiocytes of Atm-/- and control mice. We propose that the severe disruption of spermatogenesis during early prophase I in the absence of functional Atm may be partly due to altered interactions of telomeres with the nuclear matrix and distorted meiotic telomere clustering. PMID- 10373559 TI - The linker domain of Stat1 is required for gamma interferon-driven transcription. AB - Upon binding of gamma interferon (IFN-gamma) to its receptor, the latent transcription factor Stat1 becomes phosphorylated, dimerizes, and enters the nucleus to activate transcription. In response to IFN-alpha, Stat1 binds to Stat2 in a heterodimer that recruits p48, an IRF family member, to activate transcription. A number of functional domains of the STATs, including a C terminal transactivation domain, a dimerization domain, and an SH2 domain, are known. However, the highly conserved residues between the DNA binding and SH2 domains (463 to 566), recently christened the linker domain on the basis of crystallographic studies, have remained without a known function. In the present study, we report that KE544-545AA point mutants in Stat1 abolish transcriptional responses to IFN-gamma but not to IFN-alpha. We further show that this mutant Stat1 undergoes normal phosphorylation, nuclear translocation, and DNA binding. Taken together with recent structural evidence, these results suggest that the linker domain acts as a critical contact point during the construction of a Stat1 driven transcriptional complex. PMID- 10373560 TI - Overproduction of human Myt1 kinase induces a G2 cell cycle delay by interfering with the intracellular trafficking of Cdc2-cyclin B1 complexes. AB - The Myt1 protein kinase functions to negatively regulate Cdc2-cyclin B complexes by phosphorylating Cdc2 on threonine 14 and tyrosine 15. Throughout interphase, human Myt1 localizes to the endoplasmic reticulum and Golgi complex, whereas Cdc2 cyclin B1 complexes shuttle between the nucleus and the cytoplasm. Here we report that overproduction of either kinase-active or kinase-inactive forms of Myt1 blocked the nuclear-cytoplasmic shuttling of cyclin B1 and caused cells to delay in the G2 phase of the cell cycle. The COOH-terminal 63 amino acids of Myt1 were identified as a Cdc2-cyclin B1 interaction domain. Myt1 mutants lacking this domain no longer bound cyclin B1 and did not efficiently phosphorylate Cdc2 cyclin B1 complexes in vitro. In addition, cells overproducing mutant forms of Myt1 lacking the interaction domain exhibited normal trafficking of cyclin B1 and unperturbed cell cycle progression. These results suggest that the docking of Cdc2-cyclin B1 complexes to the COOH terminus of Myt1 facilitates the phosphorylation of Cdc2 by Myt1 and that overproduction of Myt1 perturbs cell cycle progression by sequestering Cdc2-cyclin B1 complexes in the cytoplasm. PMID- 10373561 TI - Failure of poly(ADP-ribose) polymerase cleavage by caspases leads to induction of necrosis and enhanced apoptosis. AB - Activation of poly(ADP-ribose) polymerase (PARP) by DNA breaks catalyzes poly(ADP ribosyl)ation and results in depletion of NAD+ and ATP, which is thought to induce necrosis. Proteolytic cleavage of PARP by caspases is a hallmark of apoptosis. To investigate whether PARP cleavage plays a role in apoptosis and in the decision of cells to undergo apoptosis or necrosis, we introduced a point mutation into the cleavage site (DEVD) of PARP that renders the protein resistant to caspase cleavage in vitro and in vivo. Here, we show that after treatment with tumor necrosis factor alpha, fibroblasts expressing this caspase-resistant PARP exhibited an accelerated cell death. This enhanced cell death is attributable to the induction of necrosis and an increased apoptosis and was coupled with depletion of NAD+ and ATP that occurred only in cells expressing caspase resistant PARP. The PARP inhibitor 3-aminobenzamide prevented the NAD+ drop and concomitantly inhibited necrosis and the elevated apoptosis. These data indicate that this accelerated cell death is due to NAD+ depletion, a mechanism known to kill various cell types, caused by activation of uncleaved PARP after DNA fragmentation. The present study demonstrates that PARP cleavage prevents induction of necrosis during apoptosis and ensures appropriate execution of caspase-mediated programmed cell death. PMID- 10373562 TI - Trimeric association of Hox and TALE homeodomain proteins mediates Hoxb2 hindbrain enhancer activity. AB - Pbx/exd proteins modulate the DNA binding affinities and specificities of Hox proteins and contribute to the execution of Hox-dependent developmental programs in arthropods and vertebrates. Pbx proteins also stably heterodimerize and bind DNA with Meis and Pknox1-Prep1, additional members of the TALE (three-amino-acid loop extension) superclass of homeodomain proteins that function on common genetic pathways with a subset of Hox proteins. In this study, we demonstrated that Pbx and Meis bind DNA as heterotrimeric complexes with Hoxb1 on a genetically defined Hoxb2 enhancer, r4, that mediates the cross-regulatory transcriptional effects of Hoxb1 in vivo. The DNA binding specificity of the heterotrimeric complex for r4 is mediated by a Pbx-Hox site in conjunction with a distal Meis site, which we showed to be required for ternary complex formation and Meis-enhanced transcription. Formation of heterotrimeric complexes in which all three homeodomains bind their cognate DNA sites is topologically facilitated by the ability of Pbx and Meis to interact through their amino termini and bind DNA without stringent half-site orientation and spacing requirements. Furthermore, Meis site mutation in the Hoxb2 enhancer phenocopies Pbx-Hox site mutation to abrogate enhancer-directed expression of a reporter transgene in the murine embryonic hindbrain, demonstrating that DNA binding by all three proteins is required for trimer function in vivo. Our data provide in vitro and in vivo evidence for the combinatorial regulation of Hox and TALE protein functions that are mediated, in part, by their interdependent DNA binding activities as ternary complexes. As a consequence, Hoxb1 employs Pbx and Meis-related proteins, as a pair of essential cofactors in a higher-order molecular complex, to mediate its transcriptional effects on an endogenous Hox response element. PMID- 10373563 TI - Mitogen-activated protein kinase kinase kinase 1 activates androgen receptor dependent transcription and apoptosis in prostate cancer. AB - Mitogen-activated protein (MAP) kinases phosphorylate the estrogen receptor and activate transcription from estrogen receptor-regulated genes. Here we examine potential interactions between the MAP kinase cascade and androgen receptor mediated gene regulation. Specifically, we have studied the biological effects of mitogen-activated protein kinase kinase kinase 1 (MEKK1) expression in prostate cancer cells. Our findings demonstrate that expression of constitutively active MEKK1 induces apoptosis in androgen receptor-positive but not in androgen receptor-negative prostate cancer cells. Reconstitution of the androgen receptor signaling pathway in androgen receptor-negative prostate cancer cells restores MEKK1-induced apoptosis. MEKK1 also stimulates the transcriptional activity of the androgen receptor in the presence or absence of ligand, whereas a dominant negative mutant of MEKK1 impairs activation of the androgen receptor by androgen. These studies demonstrate an unanticipated link between MEKK1 and hormone receptor signaling and have implications for the molecular basis of hormone independent prostate cancer growth. PMID- 10373564 TI - Defects in components of the proteasome enhance transcriptional silencing at fission yeast centromeres and impair chromosome segregation. AB - Fission yeast centromeres are transcriptionally silent and form a heterochromatin like structure essential for normal centromere function; this appears analogous to heterochromatin and position effect variegation in other eukaryotes. Conditional mutations in three genes designated cep (centromere enhancer of position effect) were found to enhance transcriptional silencing within centromeres. Cloning of the cep1(+) and cep2(+) genes by functional complementation revealed that they are identical to the previously described genes pad1(+) and mts2(+), respectively, which both encode subunits of the proteasome 19S cap. Like Mts2 and Mts4, epitope-tagged Cep1/Pad1 localizes to or near the nuclear envelope throughout the cell cycle. The cep mutants display a range of phenotypes depending on the temperature. Silencing within the central domain of centromeres is increased at 36 degrees C. This suggests that the proteasome is involved in regulating silencing and thus centromeric chromatin architecture, possibly by lowering the level of some chromatin-associated protein by ubiquitin-dependent degradation. This is the first report of defective proteasome function affecting heterochromatin-mediated transcriptional silencing. At 36 and 32 degrees C, the cep mutants lose chromosomes at an elevated rate, and at 18 degrees C, the mutants are cryosensitive for growth. Cytological analysis at 18 degrees C revealed a defect in sister chromatid separation while other mitotic events occurred normally, indicating that cep mutations might interfere specifically with the degradation of inhibitor(s) of sister chromatid separation. These observations suggest that 19S subunits confer a level of substrate specificity on the proteasome and raise the possibility of a link between components involved in centromere architecture and sister chromatid cohesion. PMID- 10373565 TI - Sister chromatid exchanges are mediated by homologous recombination in vertebrate cells. AB - Sister chromatid exchange (SCE) frequency is a commonly used index of chromosomal stability in response to environmental or genetic mutagens. However, the mechanism generating cytologically detectable SCEs and, therefore, their prognostic value for chromosomal stability in mitotic cells remain unclear. We examined the role of the highly conserved homologous recombination (HR) pathway in SCE by measuring SCE levels in HR-defective vertebrate cells. Spontaneous and mitomycin C-induced SCE levels were significantly reduced for chicken DT40 B cells lacking the key HR genes RAD51 and RAD54 but not for nonhomologous DNA end joining (NHEJ)-defective KU70(-/-) cells. As measured by targeted integration efficiency, reconstitution of HR activity by expression of a human RAD51 transgene restored SCE levels to normal, confirming that HR is the mechanism responsible for SCE. Our findings show that HR uses the nascent sister chromatid to repair potentially lethal DNA lesions accompanying replication, which might explain the lethality or tumorigenic potential associated with defects in HR or HR-associated proteins. PMID- 10373566 TI - PIC-1/SUMO-1-modified PML-retinoic acid receptor alpha mediates arsenic trioxide induced apoptosis in acute promyelocytic leukemia. AB - Fusion proteins involving the retinoic acid receptor alpha (RARalpha) and PML or PLZF nuclear protein are the genetic markers of acute promyelocytic leukemia (APL). APLs with PML-RARalpha or PLZF-RARalpha fusion protein differ only in their response to retinoic acid (RA) treatment: the t(15;17) (PML-RARalpha positive) APL blasts are sensitive to RA in vitro, and patients enter disease remission after RA treatment, while those with t(11;17) (PLZF-RARalpha-positive) APLs do not. Recently it has been shown that complete remission can be achieved upon treatment with arsenic trioxide (As2O3) in PML-RARalpha-positive APL, even when the patient has relapsed and the disease is RA resistant. This appears to be due to apoptosis induced by As2O3 in the APL blasts by poorly defined mechanisms. Here we report that (i) As2O3 induces apoptosis only in cells expressing the PML RARalpha, not the PLZF-RARalpha, fusion protein; (ii) PML-RARalpha is partially modified by covalent linkage with a PIC-1/SUMO-1-like protein prior to As2O3 treatment, whereas PLZF-RARalpha is not; (iii) As2O3 treatment induces a change in the modification pattern of PML-RARalpha toward highly modified forms; (iv) redistribution of PML nuclear bodies (PML-NBs) upon As2O3 treatment is accompanied by recruitment of PIC-1/SUMO-1 into PML-NBs, probably due to hypermodification of both PML and PML-RARalpha; (v) As2O3-induced apoptosis is independent of the DNA binding activity located in the RARalpha portion of the PML-RARalpha fusion protein; and (vi) the apoptotic process is bcl-2 and caspase 3 independent and is blocked only partially by a global caspase inhibitor. Taken together, these data provide novel insights into the mechanisms involved in As2O3 induced apoptosis in APL and predict that treatment of t(11;17) (PLZF-RARalpha positive) APLs with As2O3 will not be successful. PMID- 10373567 TI - The disabled 1 phosphotyrosine-binding domain binds to the internalization signals of transmembrane glycoproteins and to phospholipids. AB - Disabled gene products are important for nervous system development in drosophila and mammals. In mice, the Dab1 protein is thought to function downstream of the extracellular protein Reln during neuronal positioning. The structures of Dab proteins suggest that they mediate protein-protein or protein-membrane docking functions. Here we show that the amino-terminal phosphotyrosine-binding (PTB) domain of Dab1 binds to the transmembrane glycoproteins of the amyloid precursor protein (APP) and low-density lipoprotein receptor families and the cytoplasmic signaling protein Ship. Dab1 associates with the APP cytoplasmic domain in transfected cells and is coexpressed with APP in hippocampal neurons. Screening of a set of altered peptide sequences showed that the sequence GYXNPXY present in APP family members is an optimal binding sequence, with approximately 0.5 microM affinity. Unlike other PTB domains, the Dab1 PTB does not bind to tyrosine phosphorylated peptide ligands. The PTB domain also binds specifically to phospholipid bilayers containing phosphatidylinositol 4P (PtdIns4P) or PtdIns4,5P2 in a manner that does not interfere with protein binding. We propose that the PTB domain permits Dab1 to bind specifically to transmembrane proteins containing an NPXY internalization signal. PMID- 10373568 TI - Trithorax- and Polycomb-group response elements within an Ultrabithorax transcription maintenance unit consist of closely situated but separable sequences. AB - In Drosophila, two classes of genes, the trithorax group and the Polycomb group, are required in concert to maintain gene expression by regulating chromatin structure. We have identified Trithorax protein (TRX) binding elements within the bithorax complex and have found that within the bxd/pbx regulatory region these elements are functionally relevant for normal expression patterns in embryos and confer TRX binding in vivo. TRX was localized to three closely situated sites within a 3-kb chromatin maintenance unit with a modular structure. Results of an in vivo analysis showed that these DNA fragments (each approximately 400 bp) contain both TRX- and Polycomb-group response elements (TREs and PREs) and that in the context of the endogenous Ultrabithorax gene, all of these elements are essential for proper maintenance of expression in embryos. Dissection of one of these maintenance modules showed that TRX- and Polycomb-group responsiveness is conferred by neighboring but separable DNA sequences, suggesting that independent protein complexes are formed at their respective response elements. Furthermore, we have found that the activity of this TRE requires a sequence (approximately 90 bp) which maps to within several tens of base pairs from the closest neighboring PRE and that the PRE activity in one of the elements may require a binding site for PHO, the protein product of the Polycomb-group gene pleiohomeotic. Our results show that long-range maintenance of Ultrabithorax expression requires a complex element composed of cooperating modules, each capable of interacting with both positive and negative chromatin regulators. PMID- 10373569 TI - Critical role played by cyclin D3 in the MyoD-mediated arrest of cell cycle during myoblast differentiation. AB - During the terminal differentiation of skeletal myoblasts, the activities of myogenic factors regulate not only tissue-specific gene expressions but also the exit from the cell cycle. The induction of cell cycle inhibitors such as p21 and pRb has been shown to play a prominent role in the growth arrest of differentiating myoblasts. Here we report that, at the onset of differentiation, activation by MyoD of the Rb, p21, and cyclin D3 genes occurs in the absence of new protein synthesis and with the requirement of the p300 transcriptional coactivator. In differentiated myocytes, cyclin D3 also becomes stabilized and is found nearly totally complexed with unphosphorylated pRb. The detection of complexes containing cyclin D3, cdk4, p21, and PCNA suggests that cdk4, along with PCNA, may get sequestered into high-order structures held together by pRb and cyclin D3. Cyclin D3 up-regulation and stabilization is inhibited by adenovirus E1A, and this correlates with the ability of E1A to promote pRb phosphorylation; conversely, the overexpression of cyclin D3 in differentiated myotubes counteracts the E1A-mediated reactivation of DNA synthesis. These results indicate that cyclin D3 critically contributes to the irreversible exit of differentiating myoblasts from the cell cycle. PMID- 10373570 TI - Spatial organization of the core region of yeast TFIIIB-DNA complexes. AB - The interaction of yeast TFIIIB with the region upstream of the SUP4 tRNATyr gene was extensively probed by use of photoreactive phosphodiesters, deoxyuridines, and deoxycytidines that are site specifically incorporated into DNA. The TATA binding protein (TBP) was found to be in close proximity to the minor groove of a TATA-like DNA sequence that starts 30 nucleotides upstream of the start site of transcription. TBP was cross-linked to the phosphate backbone of DNA from bp -30 to -20 in the nontranscribed strand and from bp -28 to -24 in the transcribed strand (+1 denotes the start site of transcription). Most of the major groove of DNA in this region was shown not to be in close proximity to TBP, thus resembling the binding of TBP to the TATA box, with one notable exception. TBP was shown to interact with the major groove of DNA primarily at bp -23 and to a lesser degree at bp -25 in the transcribed strand. The stable interaction of TBP with the major groove at bp -23 was shown to require the B" subunit of TFIIIB. The S4 helix and flanking region of TBP were shown to be proximal to the major groove of DNA by peptide mapping of the region of TBP cross-linked at bp -23. Thus, TBP in the TFIIIB-SUP4 gene promoter region is bound in the same direction as TBP bound to the TATA box with respect to the transcription start site. The B" and TFIIB related factor (BRF) subunits of TFIIIB are positioned on opposite sides of the TBP-DNA core of the TFIIIB complex, as indicated by correlation of cross-linking data to the crystal structure of the TBP-TATA box complex. Evidence is given for BRF binding near the C-terminal stirrup of TBP, similar to that of TFIIB near the TBP-TATA box complex. The protein clamp formed around the TBP-DNA complex by BRF and B" would help explain the long half-life of the TFIIIB-DNA complex and its resistance to polyanions and high salt. The path of DNA traversing the surface of TBP at the 3' end of the TATA-like element in the SUP4 tRNA gene is not the same as that of TBP bound to a TATA box element, as shown by the cross-linking of TBP at bp -23. PMID- 10373571 TI - Bioactivity of anti-angiogenic ribozymes targeting Flt-1 and KDR mRNA. AB - Vascular endothelial growth factor (VEGF) and its receptors Flt-1 and KDR play important roles in physiological and pathological angiogenesis. Ribozymes that target the VEGF receptor mRNAs were developed and their biological activities in cell culture and an animal model were assessed. Ribozymes targeting Flt-1 or KDR mRNA sites reduced VEGF-induced proliferation of cultured human vascular endothelial cells and specifically lowered the level of Flt-1 or KDR mRNA present in the cells. Anti- Flt-1 and KDR ribozymes also exhibited anti-angiogenic activity in a rat corneal pocket assay of VEGF-induced angiogenesis. This report illustrates the anti-angiogenic potential of these ribozymes as well as their value in studying VEGF receptor function in normal and pathophysiologic states. PMID- 10373572 TI - Dissection of the NF-Y transcriptional activation potential. AB - NF-Y is a trimeric CCAAT-binding factor with histone fold subunits (NF-YB/NF-YC) and bipartite activation domains located on NF-YA and NF-YC. We reconstituted the NF-Y activation potential in vivo with GAL4 DBD fusions. In the GAL4-YA configuration, activation requires co-expression of the three subunits; with GAL4 YB and GAL4-YC, transfections of the histone fold partners are sufficient, provided that the Q-rich domain of NF-YC is present. Combinations of mutants indicate that the Q-rich domains of NF-YA and NF-YC are redundant in the trimeric complex. Glutamines 101 and 102 of NF-YA are required for activity. We assayed NF Y on different promoter targets, containing single or multiple GAL4 sites: whereas on a single site NF-Y is nearly as powerful as VP16, on multiple sites neither synergistic nor additive effects are observed. NF-Y activates TATA and Inr core elements and the overall potency is in the same range as other Q-rich and Pro-rich activation domains. These results represent the first in vivo evidence of subunit interactions studies and further support the hypothesis that NF-Y is a general promoter organizer rather than a brute activator. PMID- 10373573 TI - RNA molecules containing exons originating from different members of the cytochrome P450 2C gene subfamily (CYP2C) in human epidermis and liver. AB - Reverse transcription-PCR analysis in human epidermis, using primers from CYP2C18 and CYP2C19, revealed products containing combinations between canonically defined exons of these two genes. The major RNA species identified contained 2C18 exon 8 spliced with 2C19 exon 2. However, the terminal exons 1 and 9 were never detected in any of these composite molecules. When similar experiments were performed with liver RNA, exons 1 and 9 of both 2C18 and 2C19 were readily identified in composite 2C18/2C19 RNAs. Moreover, molecules containing 2C9 sequences spliced with 2C18 exons were also detected. These findings suggest that during the process of RNA splicing of the 2C transcripts, various exon juxtaposition events may occur, including combinations between exons of distinct genes. However, the frequency of these events is quite low and the levels of the composite RNA molecules are generally estimated at less than one molecule per cell. Since the order of these genes on chromosome 10q24 is CYP2C18 - CYP2C19 - CYP2C9, it is conceivable that the composite RNAs may result from multiple canonical and inverse splicing events of a long pre-mRNA that encompasses the three genes. However, these molecules could also be rationalized as being the products of trans splicing phenomena between distinct pre-mRNAs. PMID- 10373574 TI - Antisense display--a method for functional gene screening: evaluation in a cell free system and isolation of angiogenesis-related genes. AB - Presented here is an antisense-oriented method for functional gene screening, which we propose naming 'antisense display'. In principle, it consists of four steps: (i) preparation of phosphorothioate antisense repertoires that would correspond to the Kozak's consensus sequence, (ii) subgroup screening to identify active antisense molecules that could cause changes in the cellular phenotypes concerned and (iii) RT-PCR cloning of cDNA with the 5[prime] sense complement and 3[prime] anchor primers and sequence determination, followed by (iv) functional assays of candidate genes. Cell-free translation in rabbit reticulocyte lysate revealed that 10mer or longer antisense effectively halted protein synthesis. This required the presence of RNase H, and was achieved without prior heat denaturation of the RNA templates. Then, subpools of the 10mer repertoire were administered to human microvascular endothelial cells in culture, and screened for anti-angiogenic activities. A single species having the sequence 5[prime] GGCTCATGGT-3[prime] consistently inhibited the endothelial cell growth under hypoxia. Through RT-PCR with the corresponding sense primer, we came across three candidate cDNAs. Experiments employing longer unique antisense reproduced marked growth inhibitions in two of the three cDNAs. One encoded a mitochondrial protein and the other, which encoded a putative type-2 membrane protein containing Rab GAP/TBC and EF-hand like domains, was a gene previously undescribed in human. The results suggest that the antisense display method is potentially useful for isolating new genes towards elucidating their functions. PMID- 10373575 TI - Spermine inhibition of the 2,5-diaziridinyl-1,4-benzoquinone (DZQ) crosslinking reaction with DNA duplexes containing poly(purine). poly(pyrimidine) tracts. AB - Upon reduction, 2,5-diaziridinyl-1,4-benzoquinone (DZQ) can form an interstrand guanine to guanine crosslink with DNA duplexes containing a d(GC).d(GC) dinucleotide step. The reaction is enhanced by a thymine positioned 5[prime] to each guanine [i.e. in a d(TGCA). d(TGCA) duplex fragment]. Here we show that spermine can inhibit DZQ crosslink formation in duplexes of sequence d[C(N6)TGCA(M6)C]. d[G(M[prime]6)TG-CA(N[prime]6)G]. For N6= M6= GGGGGG, N6= M6= a 'random' sequence and N6= GGGGGG and M6= a 'random' sequence, spermine concentrations of 20, 1 and 3 microM, respectively, were required for 50% inhibition of the DZQ crosslink. This suggests that spermine is more strongly bound to the polyguanosine tract than the random sequence, making it less available for crosslink inhibition. When the polyguanosine tract is interrupted by N 7-deazaguanine (D) located three bases, d(CGGGDGGTGCAGGDGGGC), and four bases, d(CG-GDGGGTGCAGGGDGGC), from the d(TGCA).d(TGCA) site, 30 and 3 microM spermine, respectively, were required for 50% crosslink inhibition. We suggest that this difference is due to the relative proximity of the three-guanosine tract to the d(TGCA).d(TGCA) site. We were able to confirm these conclusions with further experiments using duplexes containing three-guanosine and two-guanosine tracts and from computer simulations of the spermine-DNA complexes. PMID- 10373576 TI - DNA structural transitions within the PKD1 gene. AB - Autosomal dominant polycystic kidney disease (ADPKD) affects over 500 000 Americans. Eighty-five percent of these patients have mutations in the PKD1 gene. The focal nature of cyst formation has recently been attributed to innate instability in the PKD1 gene. Intron 21 of this gene contains the largest polypurine. polypyrimidine tract (2.5 kb) identified to date in the human genome. Polypurine.polypyrimidine mirror repeats form intramolecular triplexes, which may predispose the gene to mutagenesis. A recombinant plasmid containing the entire PKD1 intron 21 was analyzed by two-dimensional gel electrophoresis and it exhibited sharp structural transitions under conditions of negative supercoiling and acidic pH. The superhelical density at which the transition occurred was linearly related to pH, consistent with formation of protonated DNA structures. P1 nuclease mapping studies of a plasmid containing the entire intron 21 identified four single-stranded regions where structural transitions occurred at low superhelical densities. Two-dimensional gel electrophoresis and chemical modification studies of the plasmid containing a 46 bp mirror repeat from one of the four regions demonstrated the formation of an H-y3 triplex structure. In summary, these experiments demonstrate that a 2500 bp polypurine.polypyrimidine tract within the PKD1 gene is capable of forming multiple non-B-DNA structures. PMID- 10373577 TI - Srp2, an SR protein family member of fission yeast: in vivo characterization of its modular domains. AB - We isolated srp2, a gene encoding a protein composed of two RNA binding domains (RBDs) at the N-terminus followed by an arginine-rich region that is flanked by two short SR (serine/arginine) elements. The RBDs contain the signatures RDADDA and SWQDLKD found in RBD1 and RBD2 of all typical metazoan SR proteins. srp2 is essential for growth. We have analyzed in vivo the role of the modular domains of Srp2 by testing specific mutations in a conditional strain for complementation. We found that RBD2 is essential for function and determines the specificity of RBD1 in Srp2. Replacement of the first RBD with RBD1 of Srp1 of fission yeast does not change this specificity. The two SR elements in the C-terminus of Srp2 are also essential for function in vivo. Cellular distribution analysis with green fluorescence protein fused to portions of Srp2 revealed that the SR elements are necessary to target Srp2 to the nucleus. Furthermore, overexpression of modular domains of Srp2 and Srp1 show different effects on pre-mRNA splicing activity of the tfIId gene. Taken together, these findings are consistent with the notion that the RBDs of these proteins may be involved in pre-mRNA recognition. PMID- 10373578 TI - A clean data set of EST-confirmed splice sites from Homo sapiens and standards for clean-up procedures. AB - A clean data set of verified splice sites from Homo sapiens are reported as well as the standards used for the clean-up procedure. The sites were validated by: (i) standard cleaning procedures such as requiring consistency in the annotation of the gene structural elements, completeness of the coding regions and elimination of redundant sequences; (ii) clustering by decision trees coupled with analysis of ClustalW alignments of the translated protein sequence with homologous proteins from SWISS-PROT; (iii) matching against human EST sequences. The sites are categorised as: (i) donor sites, a set of 619 EST-confirmed donor sites, for which 138 are either the sites or the regions around the sites involved in alternative splice events; (ii) acceptor sites, a set of 623 EST confirmed acceptor sites, for which 144 are either the sites or the regions around the sites are involved in alternative splice events; (iii) genuine splice sites, a set of 392 splice sites wherein both the donor and acceptor sites had EST confirmation and were not involved in any alternative splicing; (iv) alternative splice sites, a set of 209 splice sites wherein both the donor and acceptor sites had EST confirmation and the sites or the regions around them were involved in alternative splicing. A set of nucleotide regions that can be used to generate a control set of false splice sites that have a high confidence of being non-functional are also reported. PMID- 10373579 TI - Single amino acid substitutions in the HsdR subunit of the type IB restriction enzyme EcoAI uncouple the DNA translocation and DNA cleavage activities of the enzyme. AB - Type I restriction enzymes bind to specific DNA sequences but subsequently translocate non-specific DNA past the complex in a reaction coupled to ATP hydrolysis and cleave DNA at any barrier that can halt the translocation process. The restriction subunit of these enzymes, HsdR, contains a cluster of seven amino acid sequence motifs typical of helicase superfamily II, that are believed to be relevant to the ATP-dependent DNA translocation. Alignment of all available HsdR sequences reveals an additional conserved region at the protein N-terminus with a consensus sequence reminiscent of the P-D.(D/E)-X-K catalytic motif of many type II restriction enzymes. To investigate the role of these conserved residues, we have produced mutants of the type IB restriction enzyme Eco AI. We have found that single alanine substitutions at Asp-61, Glu-76 and Lys-78 residues of the HsdR subunit abolished the enzyme's restriction activity but had no effect on its ATPase and DNA translocation activities, suggesting that these residues are part of the active site for DNA cleavage. PMID- 10373580 TI - BseSI, a restriction endonuclease from Bacillus stearothermophilus Jo 10-553, which recognizes the novel hexanucleotide sequence 5'-G(G/T)GC(A/C)C-3'. AB - A new restriction endonuclease Bse SI has been isolated from Bacillus stearothermophilus Jo10-553. Bse SI recognizes a degenerate hexanucleotide sequence 5'-G(G/T)GC(A/C)C-3' and cleaves DNA to produce 3[prime]-protruding tetranucleotide ends. PMID- 10373581 TI - Post-translational control of the MEF2A transcriptional regulatory protein. AB - Myocyte enhancer factor 2 (MEF2) transcriptional regulatory proteins are key regulators of muscle-specific gene expression and also play a general role in the cellular response to growth factors, cytokines and environmental stressors. To identify signaling pathway components that might mediate these events, the potential role of MAP kinase and PKC signaling in the modulation of MEF2A phosphorylation and transcriptional activity were therefore studied. In transient transfection reporter assays, activated p38 MAP kinase potently increased MEF2A trans -activating potential, PKC[delta] and [epsiv] isotypes enhanced MEF2A transactivation to a lesser extent, while the ERK1/2 and JNK/SAPK pathways were without effect. A GAL4-based assay system showed that p38 MAP kinase and PKC[delta] target the MEF2A transactivation domain. We also observed an increase in p38 MAP kinase activity in congruence with the increase in MEF2A expression in differentiating primary muscle cells. COS cells overexpressing MEF2A alone or with one of the kinases were metabolically labeled with [32P]orthophosphate and MEF2A was immunoprecipitated using specific anti-MEF2A antibodies. MEF2A from cells co-transfected with activated p38 MAP kinase showed a decreased electrophoretic mobility due to phosphorylation. Subsequent phosphopeptide mapping and phosphoamino acid analysis indicated the appearance of several phoshopeptides due to p38 MAP kinase activation of MEF2A which were due to phosphorylation on serine and threonine residues. These studies position MEF2A as a nuclear target for the p38 MAP kinase signaling pathway. PMID- 10373582 TI - The role of Schizosaccharomyces pombe Rad32, the Mre11 homologue, and other DNA damage response proteins in non-homologous end joining and telomere length maintenance. AB - The Schizosaccharomyces pombe homologue of Mre11, Rad32, is required for repair of UV- and ionising radiation-induced DNA damage and meiotic recombination. In this study we have investigated the role of Rad32 and other DNA damage response proteins in non-homologous end joining (NHEJ) and telomere length maintenance in S.pombe. We show that NHEJ in S.pombe occurs by an error-prone mechanism, in contrast to the accurate repair observed in Saccharomyces cerevisiae. Deletion of the rad32 gene results in a modest reduction in NHEJ activity and the remaining repair events that occur are accurate. Mutations in two of the phosphoesterase motifs in Rad32 have no effect on the efficiency or accuracy of end joining, suggesting that the role of Rad32 protein may be to recruit another nuclease(s) for processing during the end joining reaction. We also analysed NHEJ in other DNA damage response mutants and showed that the checkpoint mutant rad3-d and two recombination mutants defective in rhp51 and rhp54 (homologues of S.cerevisiae RAD51 and RAD54, respectively) are not affected. However disruption of rad22, rqh1 and rhp9 / crb2 (homologues of the S.cerevisiae RAD52, SGS1 and RAD9 genes) resulted in increased NHEJ activity. Telomere lengths in the rad32, rhp9 and rqh1 null alleles were reduced to varying extents intermediate between the lengths observed in wild-type and rad3 null cells. PMID- 10373583 TI - C-->U editing of apolipoprotein B mRNA in marsupials: identification and characterisation of APOBEC-1 from the American opossum Monodelphus domestica. AB - The C->U editing of RNA is widely found in plant and animal species. In mammals it is a discrete process confined to the editing of apolipoprotein B (apoB) mRNA in eutherians and the editing of the mitochondrial tRNA for glycine in marsupials. Here we have identified and characterised apoB mRNA editing in the American opossum Monodelphus domestica. The apoB mRNA editing site is highly conserved in the opossum and undergoes complete editing in the small intestine, but not in the liver or other tissues. Opossum APOBEC-1 cDNA was cloned, sequenced and expressed. The encoded protein is similar to APOBEC-1 of eutherians. Motifs previously identified as involved in zinc binding, RNA binding and catalysis, nuclear localisation and a C-terminal leucine-rich domain are all conserved. Opossum APOBEC-1 contains a seven amino acid C-terminal extension also found in humans and rabbits, but not present in rodents. The opossum APOBEC-1 gene has the same intron/exon organisation in the coding sequence as the eutherian gene. Northern blot and RT-PCR analyses and an editing assay indicate that no APOBEC-1 was expressed in the liver. Thus the far upstream promoter responsible for hepatic expression in rodents does not operate in the opossum. An APOBEC-1-like enzyme such as might be involved in C->U RNA editing of tRNA in marsupial mitochondria was not demonstrated. The activity of opossum APOBEC-1 in the presence of both chicken and rodent auxiliary editing proteins was comparable to that of other mammals. These studies extend the origins of APOBEC-1 back 170 000 000 years to marsupials and help bridge the gap in the origins of this RNA editing process between birds and eutherian mammals. PMID- 10373584 TI - A new approach to the synthesis of branched and branched cyclic oligoribonucleotides. AB - The six-step synthesis of the di-triethylammonium salt of 5[prime]-O -trityl-6-N pivaloyladenosine-2[prime]-(H -phosphonate)-3'-[(2-chlorophenyl) phosphate]9 from 3', 5'- O -(1,1,3,3-tetraisopropyldisiloxan-1,3-diyl)-6-N-pivaloyla denosine1in 68% overall yield is described. Compound9is converted into a branched pentaribonucleoside tetraphosphate 24 and a branched cyclic pentaribonucleotide ('lariat') 25 by solution phase triester chemistry involving both H-phosphonate and conventional phosphotriester coupling reactions. The monomeric building block 9 is proposed as a universal synthon for the preparation of branched and branched cyclic oligoribonucleotides derived from adenosine. PMID- 10373585 TI - A comprehensive comparison of multiple sequence alignment programs. AB - In recent years improvements to existing programs and the introduction of new iterative algorithms have changed the state-of-the-art in protein sequence alignment. This paper presents the first systematic study of the most commonly used alignment programs using BAliBASE benchmark alignments as test cases. Even below the 'twilight zone' at 10-20% residue identity, the best programs were capable of correctly aligning on average 47% of the residues. We show that iterative algorithms often offer improved alignment accuracy though at the expense of computation time. A notable exception was the effect of introducing a single divergent sequence into a set of closely related sequences, causing the iteration to diverge away from the best alignment. Global alignment programs generally performed better than local methods, except in the presence of large N/C-terminal extensions and internal insertions. In these cases, a local algorithm was more successful in identifying the most conserved motifs. This study enables us to propose appropriate alignment strategies, depending on the nature of a particular set of sequences. The employment of more than one program based on different alignment techniques should significantly improve the quality of automatic protein sequence alignment methods. The results also indicate guidelines for improvement of alignment algorithms. PMID- 10373586 TI - DNA minor groove recognition of a non-self-complementary AT-rich sequence by a tris-benzimidazole ligand. AB - The crystal structure of the non-self-complementary dodecamer DNA duplex formed by d(CG[5BrC]ATAT-TTGCG) and d(CGCAAATATGCG) has been solved to 2.3 A resolution, together with that of its complex with the tris-benzimidazole minor groove binding ligand TRIBIZ. The inclusion of a bromine atom on one strand in each structure enabled the possibility of disorder to be discounted. The native structure has an exceptional narrow minor groove, of 2.5-2.6 A in the central part of the A/T region, which is increased in width by approximately 0.8 A on drug binding. The ligand molecule binds in the central part of the sequence. The benzimidazole subunits of the ligand participate in six bifurcated hydrogen bonds with A:T base pair edges, three to each DNA strand. The presence of a pair of C H...O hydrogen bonds has been deduced from the close proximity of the pyrrolidine group of the ligand to the TpA step in the sequence. PMID- 10373587 TI - Stability of G,A triple helices. AB - In this work we selected double-stranded DNA sequences capable of forming stable triplexes at 20 or 50 degrees C with corresponding 13mer purine oligonucleotides. This selection was obtained by a double aptamer approach where both the starting sequences of the oligonucleotides and the target DNA duplex were random. The results of selection were confirmed by a cold exchange method and the influence of the position of a 'mismatch' on the stability of the triplex was documented in several cases. The selected sequences obey two rules: (i) they have a high G content; (ii) for a given G content the stability of the resulting triplex is higher if the G residues lie in stretches. The computer simulation of the Mg2+, Na+and Cl-environment around three triplexes by a density scaled Monte Carlo method provides an interpretation of the experimental observations. The Mg2+cations are statistically close to the G N7 and relatively far from the A N7. The presence of an A repels the Mg2+from adjacent G residues. Therefore, the triplexes are stabilized when the Mg2+can form a continuous spine on G N7. PMID- 10373588 TI - RNA binding characteristics and overall topology of the vaccinia poly(A) polymerase-processivity factor-primer complex. AB - The vaccinia virus-encoded heterodimer responsible for poly(A) tail elongation comprises a polyadenylylation catalytic subunit (VP55) and associated processivity factor (VP39). We show that monomeric VP39's affinity for RNA homopolymers follows the hierarchy poly(I) >poly(U) >>poly(G) >poly(A) >poly(C), that the heterodimer interacts stably with 40-45 nucleotide nucleic acid segments, and that its homopolymer preference for polyadenylylation priming is comparable to the VP39 affinity hierarchy (above). For oligonucleotide ligands possessing the previously-identified (rU)2-(N)25-rU heterodimer-binding motif, the heterodimer's affinity and base-type preference are mediated via both the (rU)2and rU portions, with the greater contribution coming from (rU)2. VP39's R107 sidechain contributes to specificity at the downstream rU. Substitution of each ribouridylate of the motif with either ribothymidine or 4-thiodeoxythymidine indicated that the downstream rU interacts with both heterodimer subunits, whereas the upstream (rU)2interacts only with VP55. A 'crosslinking SELEX' approach indicated VP39-base proximity around position -10 of a 4 thioribouridine/deoxycytidine ligand pool. Upon incubating the heterodimer with a panel of identical-sequence oligonucleotides derivatized with azidophenacyl bromide at various phosphate positions, those derivatized at positions -11 to -21 photocrosslinked to both subunits in a coordinated manner. This region may therefore pass through a 'cleft' or enclosed 'channel' at the subunit interface. PMID- 10373589 TI - p300/CBP is required for transcriptional induction by interleukin-4 and interacts with Stat6. AB - Interleukin-4 (IL-4) induces tyrosine phosphorylation of the latent transcription factor Stat6, which mediates the transcriptional responses of IL-4. The transactivation domain of Stat6 has recently been mapped to the C-terminal region of Stat6. We have investigated the mechanism by which Stat6, through its transactivation domain, induces transcription. Previous studies have shown that diverse regulated transcription factors interact with coactivators such as p300 and CBP. We report that Stat6 used the interaction with p300/CBP to exert its stimulatory effects. Overexpression of p300/CBP increased IL-4-induced transcription of Stat6 activated reporter genes. The requirement of p300/CBP for Stat6-mediated transactivation is shown by coexpression of the adenovirus E1A protein. E1A repressed the IL-4-induced reporter gene activity, while mutants of E1A, which do not interact with p300/CBP, failed to block the IL-4-induced response. In addition, we found that the minimal transactivation domain of Stat6, when fused to the GAL4 DNA-binding domain, was repressed by E1A, whereas the fusion protein p300-VP16 increased the transcriptional activity. In two-hybrid protein interaction assays in mammalian cells, we mapped the interaction domain of CBP to a C-terminal region between amino acids 1850 and 2176, a region distinct from the interaction domain of CBP with Stat1, Stat2 or Stat5. Finally, we show that antibodies raised against p300 coimmunoprecipitated Stat6 and p300 from transfected COS7 cells and antibodies against Stat6 coimmunprecipitated endogenous Stat6 and CBP from Ba/F3 cells. Our data suggest that the transactivation domain of Stat6 makes contact with the basal transcription machinery by binding to p300/CBP. PMID- 10373590 TI - The nuclear export signal-dependent localization of oligonucleopeptides enhances the inhibition of the protein expression from a gene transcribed in cytosol. AB - Upon endocytosis, most oligodeoxynucleotides (ODNs) accumulate in vesicular compartments; a tiny number of them cross the vesicle membrane, reach the cytosol and by passive diffusion enter the nucleus where they are entrapped. So far, the compartment in which an antisense ODN interacts with its mRNA target has not been precisely characterized. In an attempt to answer this question, ODN-peptides were designed with the aim of maintaining them in the cytosol. This has been achieved by a short peptide sequence called the nuclear export signal (NES). Upon microinjection, ODN-NES peptide conjugates were efficiently and rapidly exported from the nucleus to the cytosol whereas ODN-peptides containing an inactive NES were found to be located in the nucleus. The inhibitory activity of antisense ODN was tested in a system allowing the specific transcription of a luciferase reporter gene in the cytosol. Antisense propynylated ODN-NES peptide conjugates, directed against the luciferase gene, efficiently inhibited (75%) the cytosolic expression of luciferase whereas at the same concentration the peptide-free propynylated ODN or the propynylated ODN-peptides containing an inactive NES were nearly inactive. PMID- 10373591 TI - Enzymatic and antisense effects of a specific anti-Ki-ras ribozyme in vitro and in cell culture. AB - Due to their mode of action, ribozymes show antisense effects in addition to their specific cleavage activity. In the present study we investigated whether a hammerhead ribozyme is capable of cleaving mutated Ki-ras mRNA in a pancreatic carcinoma cell line and whether antisense effects contribute to the activity of the ribozyme. A 2[prime]-O-allyl modified hammerhead ribozyme was designed to cleave specifically the mutated form of the Ki- ras mRNA (GUU motif in codon 12). The activity was monitored by RT-PCR on Ki- ras RNA expression by determination of the relative amount of wild type to mutant Ki-ras mRNA, by 5-bromo-2[prime] deoxy-uridine incorporation on cell proliferation and by colony formation in soft agar on malignancy in the human pancreatic adenocarcinoma cell line CFPAC-1, which is heterozygous for the Ki-ras mutation. A catalytically inactive ribozyme was used as control to differentiate between antisense and cleavage activity and a ribozyme with random guide sequences as negative control. The catalytically active anti-Ki-ras ribozyme was at least 2-fold more potent in decreasing cellular Ki-ras mRNA levels, inhibiting cell proliferation and colony formation in soft agar than the catalytically inactive ribozyme. The catalytically active anti-Ki-ras ribozyme, but not the catalytically inactive or random ribozyme, increased the ratio of wild type to mutated Ki-ras mRNA in CFPAC-1 cells. In conclusion, both cleavage activity and antisense effects contribute to the activity of the catalytically active anti-Ki-ras hammerhead ribozyme. Specific ribozymes might be useful in the treatment of pancreatic carcinomas containing an oncogenic GTT mutation in codon 12 of the Ki-ras gene. PMID- 10373592 TI - Characterization of a Leishmania donovani gene encoding a protein that closely resembles a type IB topoisomerase. AB - In order to clone the gene encoding a type I DNA topoisomerase from Leishmania donovani, a PCR-amplified DNA fragment obtained with degenerate oligodeoxyribonucleotides was used to screen a genomic library from this parasite. An open reading frame of 1905 bases encoding a putative protein of 635 amino acid residues was isolated. A substantial part of the protein shares a significant degree of homology with the sequence of other known members of the IB topoisomerase family, in a highly conserved region of these enzymes termed the core domain. However, homology is completely lost after this conserved central core. Moreover, no conventional active tyrosine site could be identified. In fact, the protein expressed in Escherichia coli did not show any relaxation activity in vitro and was unable to complement a mutant deficient in topoisomerase I activity. The results of Southern blot experiments strongly suggested that the cloned gene was not a pseudogene. Northern analysis revealed that the gene was transcribed in its full length and also excluded the possibility that some form of splicing is necessary to produce a mature messenger. Furthermore, our results indicate that the gene is preferentially expressed in actively growing L.donovani promastigotes and that it is also expressed in other kinetoplastid parasites. PMID- 10373593 TI - ATP hydrolysis activity of the DEAD box protein Rok1p is required for in vivo ROK1 function. AB - The yeast ROK1 gene has been initially identified as a high copy plasmid suppressor of the kem1 null mutation and implicated in microtubule-mediated functions. Based on the deduced amino acid sequence of the ROK1 gene, Rok1p has been classified in the DEAD protein family of ATP-dependent RNA helicases. A subsequent report has suggested that Rok1p is required for rRNA processing. We report here the first study on the biochemical activity associated with Rok1p. The MBP-Rok1 hybrid protein was synthesized in Escherichia coli and purified by amylose affinity column and ion exchange chromatography. Rok1p has ATP hydrolysis activity. The significance of the conserved ATPase domains was addressed by generating a series of amino acid substitution mutations in these domains. Both in vivo lethality tests of the mutations and biochemical characterization of the mutant proteins suggest that ATP hydrolysis activity of Rok1p is essential for ROK1 function. The ATPase activity of Rok1p appears to be independent of single stranded RNA. Furthermore, replacement of the first Arg in the HRIGR domain, the known RNA-binding domain, with Thr, Ile or Lys has no detectable effect on in vivo ROK1 function. The lack of RNA dependency and some of the mutational phenotypes of ROK1 differentiate this gene from other members of the family. PMID- 10373594 TI - SELEX and missing phosphate contact analyses reveal flexibility within the AP 2[alpha] protein: DNA binding complex. AB - The AP-2 family of transcription factors are defined by the presence of a novel DNA binding domain, termed a 'basic helix-span-helix' motif. The AP-2 genes regulate important aspects of vertebrate embryogenesis and have also been linked to the control of cell proliferation and tumorigenesis, but the cellular targets that the AP-2 proteins control are largely undefined. In particular, since only a limited number of sequences have previously been utilized to define the nature of the AP-2 binding site, the range of DNA sequences recognized by the AP-2 proteins remains unknown. We have therefore utilized a SELEX analysis to identify multiple new AP-2[alpha] binding sites. Moreover, we have devised a novel missing phosphate and nucleotide competition analysis to characterize the residues in the binding site required for AP-2[alpha] protein:DNA contact. These studies suggest that the AP-2[alpha] protein:DNA complex is flexible and indicate that AP 2[alpha] can bind three related sequence motifs: GCC N3 GGC, GCC N4 GGC and GCC N3/4 GGG. The availability of these more refined consensus sequences should assist in the identification of target genes for this critical transcription factor. PMID- 10373595 TI - Long 5' leaders inhibit removal of a 3' trailer from a precursor tRNA by mammalian tRNA 3' processing endoribonuclease. AB - Mammalian tRNA 3' processing endoribonuclease (3' tRNase) can remove a 3' trailer from various pre-tRNAs without 5' leader nucleotides. To examine how 5[prime] leader sequences affect 3' processing efficiency, we performed in vitro 3' processing reactions with purified pig 3' tRNase and pre-tRNAArgs containing a 13 nt 3' trailer and a 5[prime] leader of various lengths. The 3' processing was slightly stimulated by 5[prime] leaders containing up to 7 nt, whereas leaders of 9 nt or longer severely inhibited the reaction. Structure probing indicated that the 5' leader sequences had little effect on pre-tRNA folding. Similar results were obtained using pre-tRNA(Val)s containing a 5' leader of various lengths. We also investigated whether 3'tRNase can remove 3' trailers that are stably base paired with 5' leaders to form an extended acceptor stem. Even such small 5' leaders as 3 and 6 nt, when base-paired with a 3' trailer, severely hindered removal of the 3' trailer by 3' tRNase. PMID- 10373596 TI - An efficient and accurate integration of mini-Mu transposons in vitro: a general methodology for functional genetic analysis and molecular biology applications. AB - Transposons are mobile genetic elements and have been utilized as essential tools in genetics over the years. Though highly useful, many of the current transposon based applications suffer from various limitations, the most notable of which are: (i) transposition is performed in vivo, typically species specifically, and as a multistep process; (ii) accuracy and/or efficiency of the in vivo or in vitro transposition reaction is not optimal; (iii) a limited set of target sites is used. We describe here a genetic analysis methodology that is based on bacteriophage Mu DNA transposition and circumvents such limitations. The Mu transposon tool is composed of only a few components and utilizes a highly efficient and accurate in vitro DNA transposition reaction with a low stringency of target preference. The utility of the Mu system in functional genetic analysis is demonstrated using restriction analysis and genetic footprinting strategies. The Mu methodology is readily applicable in a variety of current and emerging transposon-based techniques and is expected to generate novel approaches to functional analysis of genes, genomes and proteins. PMID- 10373598 TI - An in vitro screening technique for DNA polymerases that can incorporate modified nucleotides. Pseudo-thymidine as a substrate for thermostable polymerases. AB - DNA polymerases are desired that incorporate modified nucleotides into DNA with diminished pausing, premature termination and infidelity. Reported here is a simple in vitro assay to screen for DNA polymerases that accept modified nucleotides based on a set of primer extension reactions. In combination with the scintillation proximity assay (SPA[trade]), this allows rapid and simple screening of enzymes for their ability to elongate oligonucleotides in the presence of unnatural nucleotides. A proof of the concept is obtained using pseudo-thymidine (psiT), the C-nucleoside analog of thymidine, as the unnatural substrate. The conformational properties of psiT arising from the carbon-carbon bond between the sugar and the base make it an interesting probe for the importance of conformational restraints in the active site of polymerases during primer elongation. From a pool of commercially available thermostable polymerases, the assay identified Taq DNA polymerase as the most suitable enzyme for the PCR amplification of oligonucleotides containing psiT. Subsequent experiments analyzing PCR performance and fidelity of Taq DNA polymerase acting on psiT are presented. This is the first time that PCR has been performed with a C-nucleoside. PMID- 10373597 TI - Interaction of the nuclear protein CBF1 with the kappaB site of the IL-6 gene promoter. AB - The nuclear protein CBF1 has been shown to function as an intermediate to target transcription factors,such as the activated Notch receptor,to specific DNA sites. In this paper,we show that CBF1 from cell lines of different origin is able to bind to the[kappa]B site of the IL-6 promoter. By transfection analyses performed in HeLa cells,we demonstrate that overexpressed CBF1 acts as a negative regulator of IL-6 gene transcription and is unable to elicit Notch-dependent activation of this gene. Analyses of protein-DNA interactions indicate that the topology of the complex formed by CBF1 and the target DNA is subtly affected by sequencessurrounding the recognition site. Furthermore,we show that CBF1 induces DNA bending. This finding suggests that CBF1 may influence IL-6 gene transcription by determining a specific conformation of the promoter region. PMID- 10373599 TI - Characterisation of the adenovirus preterminal protein and its interaction with the POU homeodomain of NFIII (Oct-1). AB - Formation of the preinitiation complex for adenovirus DNA replication involves the incoming preterminal protein-adenovirus DNA polymerase heterodimer being positioned at the origin of replication by protein-DNA and protein-protein interactions. Preterminal protein directly binds to the cellular transcription factor nuclear factor III (Oct-1), via the POU homeodomain. Co-precipitation of POU with individual domains of preterminal protein expressed by in vitro translation indicated that POU contacts multiple sites on preterminal protein. Partial proteolysis of preterminal protein in the presence or absence of POU homeodomain demonstrated that many sites accessible to proteases in free preterminal protein were resistant to cleavage in the presence of POU homeodomain. The accessibility of sites in free preterminal protein to cleavage by trypsin was strongly dependent on the ionic strength, suggesting that preterminal protein may undergo a sodium chloride-induced conformational change. It is therefore likely that the POU homeodomain contacts a number of sites on preterminal protein to induce a conformational change which may influence the initiation of adenovirus DNA replication. PMID- 10373600 TI - Peptide nucleic acid (PNA) binding-mediated induction of human gamma-globin gene expression. AB - Peptide nucleic acids (PNAs) can bind to homopurine/homopyrimidine sequences of double-stranded DNA targets in a sequence-specific manner and form [PNA]2/DNA triplexes with single-stranded DNA D-loop structures at the PNA binding sites. These D-loop structures have been found to have a capacity to initiate transcription in vitro. If this strategy can be used to induce transcription of endogenous genes, it may provide a novel approach for gene therapy of many human diseases. Human [beta] globin disorders such as sickle cell anemia and beta thalassemia are very common genetic diseases that are caused by mutations in the beta-globin gene. When gamma-globin genes are highly expressed in sickle cell patients, the presence of high levels of fetal hemoglobin (HbF, alpha2gamma2) can compensate for the defective beta-globin gene product and such patients have much improved symptoms or are free of disease. However, the gamma-globin genes are developmentally regulated and normally expressed at very low levels (>1%) in adult blood cells. We have investigated the possibility of inducing gamma-globin gene expression with PNAs. Using PNAs designed to bind to the 5' flanking region of the gamma-globin gene, induction of expression of a reporter gene construct was demonstrated both in vitro and in vivo. More importantly, PNA-mediated induction of endogenous gamma-globin gene expression was also demonstrated in K562 human erythroleukemia cells. This result suggests that induction of gamma globin gene expression with PNAs might provide a new approach for the treatment of sickle cell disease. PNA-induced gene expression strategy also may have implications in gene therapy of other diseases such as genetic diseases, cancer and infectious diseases. PMID- 10373601 TI - Characterization of the effects of Escherichia coli replication terminator protein (Tus) on transcription reveals dynamic nature of the tus block to transcription complex progression. AB - We have characterized the blocks to progression of T7 and T3 RNA polymerase transcription complexes created when a Tus protein is bound to the template. The encounter with Tus impedes the progress of the transcription complexes of either enzyme. The duration of the block depends on which polymerase is used and the orientation of Tus on the DNA. Both genuine termination (dissociation of the transcription complex) and halting followed by continued progression after the block is abrogated are observed. The fraction of complexes that terminates depends on which polymerase is used and on the orientation of the Tus molecule. The efficiency of the block to transcription increases as the Tus concentration is increased, even if the concentration of Tus is already many times in excess of what is required to saturate its binding sites on the template in the absence of transcription. The block to transcription is rapidly abrogated if an excess of a DNA containing a binding site for Tus is added to a transcription reaction in which Tus and template have been preincubated. Finally, we find that transcription will rapidly displace Tus from a template under conditions that generate persistent blocks to transcription. These observations reveal that during the encounter with the transcription complex Tus rapidly dissociates from the template but that at sufficiently high concentrations Tus usually rebinds before the transcription complex can move forward. The advantage of a mechanism which can create a persistent block to transcription or replication complex progression, which can nevertheless be rapidly abrogated in response to down regulation of the blocking protein, is suggested. PMID- 10373602 TI - A novel in vivo assay for the analysis of protein-protein interaction. AB - The Ras Recruitment System (RRS) is a method for identification and isolation of protein-protein interaction. The method is based on translocation of cytoplasmic mammalian Ras protein to the inner leaflet of the plasma membrane through protein protein interaction. The system is studied in a temperature-sensitive yeast strain where the yeast Ras guanyl nucleotide exchange factor is inactive at 36 degrees C. Protein-protein interaction results in cell growth at the restrictive temperature. We developed a gene reporter assay for the analysis of protein protein interaction in mammalian cells. Ras activation in mammalian cells induces the mitogen-activated kinase cascade (MAPK), which can be monitored using Ras dependent reporter genes. This greatly extends the usefulness of the system and provides a novel assay for protein-protein interaction in mammalian cells. PMID- 10373603 TI - MARINE BIOTECHNOLOGY. PMID- 10373604 TI - Safety Evaluation of Transgenic Tilapia with Accelerated Growth. AB - Recent advances in modern marine biotechnology have permitted the generation of new strains of economically important fish species through the transfer of growth hormone genes. These transgenic fish strains show improved growth performance and therefore constitute a better alternative for aquaculture programs. Recently, we have obtained a transgenic tilapia line with accelerated growth. However, before introducing this line into Cuban aquaculture, environmental and food safety assessment was required by national authorities. Experiments were performed to evaluate the behavior of transgenic tilapia in comparison to wild tilapia as a way to assess the environmental impact of introducing transgenic tilapia into Cuban aquaculture. Studies were also conducted to evaluate, according to the principle of substantial equivalence, the safety of consuming transgenic tilapia as food. Behavior studies showed that transgenic tilapia had a lower feeding motivation and dominance status than controls. Food safety assessment indicated that tilapia growth hormone has no biological activity when administered to nonhuman primates. Furthermore, no effects were detected in human healthy volunteers after the consumption of transgenic tilapia. These results showed, at least under the conditions found in Cuba, no environmental implications for the introduction of this transgenic tilapia line and the safety in the consumption of tiGH-transgenic tilapia as an alternative feeding source for humans. These results support the culture and consumption of these transgenic tilapia. PMID- 10373605 TI - Construction of Plasmid Vectors and Transformation of the Marine Yeast Debaryomyces hansenii. AB - : We have constructed two plasmid vectors (pMR95 and pMR96) with selectable markers for the marine yeast Debaryomyces hansenii. Plasmid pMR95 contains an autonomously replicating sequence previously isolated from Debaryomyces and a hygromycin B resistance gene from the plasmid pLG90 under the control of the isocytochrome C1 promoter and terminator sequences, while pMR96 has, in addition, the Saccharomyces URA3 gene. Transformation in Debaryomyces was accomplished by electroporation. Plasmid pMR95 was capable of transforming both Saccharomyces cerevisiae and D. hansenii to hygromycin resistance at low frequencies; pMR96 transformed both yeasts at low frequencies when selected for hygromycin B resistance and at very high efficiencies when selected for uracil prototrophy. The presence of the plasmids in the transformed yeast was confirmed by polymerase chain reaction. The plasmids could be recovered back in Escherichia coli when transformed with total DNA from the yeast transformants, indicating at least a partial autonomous existence of the plasmids in the marine yeast. To our knowledge this is the first successful attempt to transform D. hansenii. PMID- 10373606 TI - Comparative Analysis of a Mitochondrial DNA Control Region Fragment Amplified from Three Adriatic Flatfish Species and Molecular Phylogenesis of Pleuronectiformes. AB - : The 5'-end of the mitochondrial control region of three Pleuronectiformes from the Adriatic Sea, Platichthys flesus italicus (Adriatic flounder), Solea vulgaris (common sole), and Solea kleini (Klein's sole), was sequenced and compared with that of six other flatfish species from the families Pleuronectidae and Bothidae. The sequence structures of all flatfishes appear very similar and consist of alternate short segments with low, medium, and high rates of nucleotide substitution. Four conserved 19-bp repeats occur at the beginning of the European and Adriatic flounder sequences. The common occurrence of tandem arrays in fish control regions could be related to a stable secondary structure. Molecular phylogenetic relationships among Pleuronectiformes agree well with previous morphologic data at all taxonomic levels. Molecular analyses could therefore contribute to resolving phylogenetic and taxonomic debates within the Pleuronectiformes. PMID- 10373607 TI - Expression of Human Glucose Transporter Type 1 and Rat Hexokinase Type II Complementary DNAs in Rainbow Trout Embryos: Effects on Glucose Metabolism. AB - : Sugars are utilized poorly in fish mainly because of low rates of transport across plasma membrane and phosphorylation. To evaluate whether it is possible to augment carbohydrate metabolism in fish using heterologous genes, expression of human glucose transporter type 1 (hGLUT1) and rat hexokinase type II (rHKII) complementary DNAs cloned with cytomegalovirus promoter was followed in rainbow trout embryos. Both genes were transcribed. Hexokinase activity, undetectable in control, was found in transformed blastulas. Increased rates of 14C-methylglucose uptake and sensitivity to cytochalasin B indicated the presence of facilitative hexose transport due to hGLUT1 expression. Effect of hGLUT1 on production of 14CO2 from glucose was greater than that of rHKII. Coexpression of the genes did not increase the rate of glucose oxidation compared with expression of hGLUT1 alone. PMID- 10373608 TI - Purification and Characterization of alpha1-Proteinase Inhibitor from Carp (Cyprinus carpio) Serum. AB - : alpha1-Proteinase inhibitor was purified to homogeneity from carp serum with an increase in specific inhibitory activity of 17-fold and a 3% recovery rate. The inhibitor was estimated to have molecular weight of 55,000 under reducing and nonreducing conditions, indicating its composition of a single polypeptide. The inhibitor immunologically crossreacted faintly with carp muscular serine proteinase inhibitor but had no crossreactivity with serine proteinase inhibitors from other species. Carp serum inhibitor exhibited marked stability over broad pH ranges of 4.0 to 10.0 and temperatures below 55 degrees C. The inhibitor potently inhibited the activities of carp intestinal and fish myofibril-binding proteinases, and its respective inhibitions of trypsin-type and carp muscular proteinases were more severe than those of chymotrypsin-type and white croaker muscular proteinases. Its inhibitions were similar to those of bovine pancreatic trypsin and alpha-chymotrypsin, and the amount required to completely inactivate 0.2 ug of each of these two proteinases was evaluated as 0.43 to 0.45 ug. This indicates a molar ratio close to 1:1 during combination of the inhibitor with each proteinase. In addition, its ability to form irreversible complexes with the proteinases was observed electrophoretically and immunologically under denaturing and reducing conditions. PMID- 10373609 TI - Callinectin, an Antibacterial Peptide from Blue Crab, Callinectes sapidus, Hemocytes. AB - This paper describes the isolation of an approximately 3.7 kDa, basic, antibacterial peptide (designated callinectin), which represents the major antibiotic activity in blue crab, Callinectes sapidus, hemocytes. A single-step purification using low-pressure cation-exchange chromatology yielded a highly purified (>95%) peptide. Purity was confirmed by C4 reverse-phase high performance liquid chromatography (RP-HPLC), native gel electrophoresis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), capillary electophoresis, and mass spectral analysis. The partial amino acid sequence obtained via Edman degradation revealed no significant homology to other reported peptides in the Basic Local Alignment Search Tool (BLAST) program database. PMID- 10373610 TI - The Benefit of a Roseobacter Species on the Survival of Scallop Larvae. AB - : A marine strain (BS107), identified as a Roseobacter species, was antagonistic to Vibrio species on agar plates. Results suggested that the inhibitory effect was displayed only in the presence of another bacterium. Quantification of the antibacterial activity showed that 48-hour-coculture supernatants from BS107 and another bacterial strain (V. anguillarum 408) reached the highest titers of bacterial inhibition. The antibacterial substance was also liberated when supernatants from V. anguillarum 408 were added to pure cultures of the inhibition-productive bacterium. The presence of a proteinaceous molecule may induce BS107 to display the inhibitory effect. The antibacterial substance was sensitive to trypsin (8000 U/ml) and stable at 100 degrees C. Cell extracts of the isolate BS107 (10(6) cells/ml) significantly enhanced scallop larval survival, thus being beneficial to the rearing process. PMID- 10373611 TI - Tc1-Like Transposable Elements in the Genome of Lake Trout (Salvelinus namaycush). AB - This study reports on the DNA sequence of a Tc1-like transposable element Tsn1 from lake trout (Salvelinus namaycush). Tc1-like elements were amplified by PCR using an oligonucleotide primer based on the Tdr1 element of zebrafish. One full length and two partial-length copies of the transposon were sequenced. In addition, partial Tsn1 elements were recovered from PCR reactions run with primers specific to the 3' terminus of the 28S rDNA. However, sequence analysis of cloned fragments found that these sequences were not associated with the rDNA cistron. Sequence comparisons indicate that Tsn1 is a type A element common to both salmonid and cyprinid fishes. The consensus sequence of the full-length element (Tsn1) was 1643 nucleotides with long terminal repeats (LTRs) of 225 nucleotides. Tsn1 contains a transposase coding region corresponding to 340 amino acids that includes all of the functional elements of Tc1-like transposons. Southern analysis found a high proportion of the Tsn1 transposons in the lake trout genome to be full-length copies. PMID- 10373612 TI - Antiviral Activities of Marine Pseudomonas Polysaccharides and Their Oversulfated Derivatives. AB - : A marine Pseudomonas species WAK-1 strain simultaneously produces extracellular glycosaminoglycan and sulfated polysaccharide. Among the antiviral activities tested for these polysaccharides, the latter showed anti-HSV-1 activity in RPMI 8226 cells (50% effective concentration is 1.4 ug/ml). Oversulfated derivatives of these polysaccharides prepared by dicyclohexylcarbodiimide-mediated reaction for both polysaccharides showed antiviral activities against influenza virus type A (for glycosaminoglycan, 50% effective concentration is 11.0 ug/ml; for another, 2.9 ug/ml). Glycosaminoglycan, sulfated polysaccharide, and their chemically synthesized oversulfated derivatives did not show antiviral activities against influenza virus type B and human immunodeficiency virus type 1. No cytotoxicity of these products was noted against host cells at the 50% cytotoxic concentration of 100 ug/ml, except that naturally occurring sulfated polysaccharide had 50% cytotoxicity against MT-4 cells at 8-21 ug/ml. PMID- 10373613 TI - Purification and Characterization of the Glutathione-S-transferases from the Northern Quahog Mercinaria mercinaria. AB - : Glutathione-S-transferase (GST) was isolated from the northern hardshell clam Mercinaria mercinaria (quahog) using a two-step procedure involving ammonium sulfate precipitation and affinity chromatography. Kinetic analysis of the purified enzyme using 1-chloro-2,4-dinitrobenzene as substrate revealed a specific activity of 38.0 umol min-1 mg-1, while Vmax and Km values were estimated as 48.0 umol min-1 mg-1 and 0.24 mM, respectively. Electrophoretic analysis of GST indicated multiple forms of the dimeric enzyme in quahogs with subunit molecular masses of 22, 24, 25, and 27 kDa. Isoelectric focusing analysis resulted in pI values for three isoenzymes of 5.1, 4.9, and 4.6. The acidic pI values obtained indicated that quahog GST belongs to the pi class. Inhibition of quahog GST by tetrapyrroles was similar to that of GST from oyster and rat liver. Quantitative comparison of tetrapyrrole inhibition patterns of quahog GST with those of oyster and rat liver GST indicated lower inhibition rates by three of the four tetrapyrroles tested (bilirubin, biliverdin, and chlorophillyin), suggesting that quahog GST could differ structurally or functionally from oyster and rat liver GSTs. PMID- 10373614 TI - Purification and Characterization of Serum Serpin from Carp (Cyprinus carpio). AB - : A serine proteinase inhibitor, termed serpin62, was purified to homogeneity from carp serum with an increase in specific inhibitory activity of 6.2-fold and a 3% recovery rate after separation from alpha1-antitrypsin. Specific inhibitory activity of serpin62 against bovine pancreatic trypsin was less than half of the specific antitryptic activity of alpha1-antitrypsin. Under both reducing and nonreducing conditions, serpin62 was estimated to have a molecular weight (62,000) apparently larger than that of alpha1-antitrypsin (55,000). They both consist of single polypeptide chains, but serpin62 differs from serine proteinase inhibitors from muscles of carp and white croaker in molecular weight and structure. Antibody raised against serpin62 immunologically crossreacted with serpin62 and had no crossreactivity with fish serum alpha1-antitrypsin and muscular analogues. The antibody was susceptible to both serpin62 and its derivatives, which were widely distributed in carp tissues. Serpin62 is most likely distinct from other fish serine proteinase inhibitors expressing antitryptic activity physicochemically and immunologically. PMID- 10373615 TI - Examination of the Montastraea annularis Species Complex (Cnidaria: Scleractinia) Using ITS and COI Sequences. AB - : The Caribbean coral Montastraea annularis has recently been proposed to be a complex of at least three sibling species. To test the validity of this proposal, we sequenced the ITS region of the nuclear ribosomal RNA gene family (ITS-1, 5.8S, and ITS-2), and a portion of the mitochondrial DNA gene cytochrome c oxidase subunit I (COI) from the three proposed species (M. annularis, M. faveolata, and M. franksi) from Florida reefs. The ITS fragment was 665 nucleotides long and had 19 variable sites, of which 6 were parsimony-informative sites. None of these sites was fixed within the proposed species. The COI fragment was 658 nucleotides long with only two sites variable in one individual. Thus, under both the biological species concept and the phylogenetic species concept, the molecular evidence gathered in this study indicates the Montastraea annularis species complex to be a single evolutionary entity as opposed to three distinct species. The three proposed Montastraea species can interbreed, ruling out prezygotic barriers to gene flow (biological species concept), and the criterion of monophyly is not satisfied if hybridization is occurring among taxa (phylogenetic species concept). PMID- 10373616 TI - Convenient Assay for Settlement Inducing Substances of Barnacles. AB - A convenient assay method for estimation of barnacle, Balanus amphitrite, settlement inductive activity was developed. To avoid the inductive effect by comrades and laborious observation requirements, cyprid larvae were put individually into wells of a 96-well plate. The settlement ratio from the experiment without any inducers was quite low; therefore this assay allowed easy estimation of settlement inductive activities. Some known inductive agents, such as serotonin and barnacle extracts, clearly showed inductive activities. This assay method is proven to be suitable for estimation of barnacle settlement inducing activities of both water-soluble and -insoluble compounds. PMID- 10373617 TI - Revelation of Antiviral Activities by Artificial Sulfation of a Glycosaminoglycan from a Marine Pseudomonas. AB - : Sulfated derivatives of a glycosaminoglycan containing l-glutamic acid produced by a marine Pseudomonas species, No. 42 strain, were prepared by the method of dicyclohexyl-carbodiimide-mediated reaction. Both low and high degrees of sulfation of the polysaccharides (products A1 and A2, respectively) were investigated for their antiviral activities against influenza virus type A (FluV A) and B (FluV-B) in MDCK cells. Both preparations showed antiviral activity against FluV-A at the 50% antiviral effective concentration of 17.3 and 5.2 ug/ml, respectively, whereas they had no antiviral activity against FluV-B. No cytotoxicity of either product was noted against MDCK cells at the 50% cytotoxic concentration of 100 ug/ml. PMID- 10373618 TI - Requisite Morphologic Interaction for Attachment between Ulva pertusa (Chlorophyta) and Symbiotic Bacteria. AB - : In order to understand the morphogenesis-inducing mechanism of Ulva pertusa by symbiotic bacteria, we observed the requisite conditions of bacteria for attachment to U. pertusa for algal morphogenesis. Non-morphogenesis-inducing bacterial mutants derived by ultraviolet irradiation did not attach onto the surface of this alga. Scanning electron microscopic observation during the process of morphogenesis in U. pertusa revealed a network-like structure formed on the algal surface within 1 week after application of bacteria. The bacteria attached onto the alga after 2 weeks of incubation. After this attachment process the morphologic change was observed in U. pertusa. PMID- 10373619 TI - Mitochondrial DNA Diversity in Three Populations of the Giant Tiger Shrimp Penaeus monodon. AB - : Mitochondrial DNA restriction fragment length polymorphism (mtDNA-RFLP) was utilized for determination of genetic variation and population structure in Penaeus monodon collected from Satun (the Andaman Sea) and Surat and Trat (the Gulf of Thailand). Twenty-eight composite haplotypes were generated from 52 restriction profiles of P. monodon mtDNA digested with 11 restriction endonucleases. The size of the entire P. monodon mitochondrial genome was estimated to be 15.913 +/- 0.177 kb. The average haplotype diversity in P. monodon was 0.864, whereas the mean nucleotide diversity within populations was 2.51%, 2.22%, and 1.91% for Satun, Trat, and Surat, respectively. Geographic heterogeneity analysis indicated population differentiation between P. monodon from the Andaman Sea and P. monodon from the Gulf of Thailand (p <.0001). On the basis of the high genetic diversity level of P. monodon in Thailand, the Satun and Trat P. monodon populations from the west and east of the pennisula were selected to be founder stocks in our selective breeding program. PMID- 10373620 TI - Characterization of a Highly Repetitive Sequence Conserved Among the North American Morone Species. AB - : A highly repetitive DNA sequence family from the genome of the North American Morone has been cloned and characterized. This family, first identified as a HindIII repetitive element, is composed of repeat units that range from 285 to 288 bp in length and comprise approximately 5.5% of the genome. The copy number of the repeat was estimated to be 1.85 x 10(5) per haploid genome set. Data from Southern blot analyses demonstrated that the HindIII repetitive element was tandemly organized. Sequence analysis of six cloned repeat monomers from each of the four North American Morone species, M. saxatilis, M. chrysops, M. americana, and M. mississippiensis, revealed a high degree of conservation of the monomeric unit. The intraspecific sequence variation ranged from 3.2% to 5.4%. A similar level of variation was detected between cloned monomers from the same individual, suggesting that most of the intraspecies variation may be due to variation among copies of the repeat. The interspecific sequence variation ranged from less than 4.6% between M. americana and M. mississippiensis to approximately 16% between the other Morone species pairs. Phylogenetic analysis of the repetitive element nucleotide sequences indicated that M. americana and M. mississippiensis were more closely related to each other than to any other pairs of Morone species. In addition, we reconstructed the Morone phylogeny using 22 previously described morphologic characters. Congruent relationships were obtained between both sets of data. The data suggest that the genus Morone is composed of two sets of sister taxa, M. saxatilis:M. chrysops and M. americana:M. mississippiensis. PMID- 10373621 TI - Complementary DNA Encoding nm23/NDP Kinase Gene from the Korean Tiger Shark Scyliorhinus torazame. AB - : A new tumor suppressor gene, snm23, homologous to the gene for human nucleoside diphosphate kinase nm23/NDP was first cloned from Korean tiger shark (Scyliorhinus torazame) skin lambda ZAP-II complementary DNA library. The gene (named snm23) containing the tumor metastasis suppressor protein was sequenced. The nucleotide and deduced amino acid sequences of snm23 revealed an open reading frame of 450 bp that corresponded to a protein of 150 amino acid residues, with a calculated molecular mass of 16.8 kDa. Sequence comparison of snm23 with nm23/NDP kinases was performed. In order to determine tissue specificity, reverse transcription-polymerase chain reaction was used. The expression of snm23/NDP kinase was detected in tissues from skin, cartilage, and liver of Korean tiger shark. PMID- 10373622 TI - Stock Identification of Gag, Mycteroperca microlepis, Along the Southeast Coast of the United States. AB - : The gag grouper Mycteroperca microlepis is an important component of commercial and recreational fisheries along the South Atlantic coast of the United States and in the Gulf of Mexico. Over the past two decades, this species has experienced significant declines in abundance and an increasing skew in sex ratios. Analysis of microsatellite DNA variation in this species shows mosaic patterns of population subdivision and significant departures from Hardy-Weinberg equilibrium in all sampling locations. Given the length of the pelagic stage (egg and larvae), the prevailing current patterns, and the migratory capabilities of the adults, it is unlikely that these observations are the result of restricted gene flow among genetically differentiated populations. The apparent structure of gag populations most likely reflects inbreeding in size-limited populations. Population declines, skewed sex ratios, and perhaps variance in female fecundity appear to have acted in concert to limited the number of individuals that contribute to a given year class. These data are reinforced by studies of other fish stocks that have experienced precipitous declines over the past two decades. PMID- 10373623 TI - A PCR-ELISA Method for Direct Detection of the Oyster Pathogen Haplosporidium nelsoni. AB - : A rapid method, utilizing both polymerase chain reaction (PCR) and enzyme linked immunosorbent assay (ELISA), was developed for detection of oyster MSX disease. The technique included using Haplosporidium nelsoni pathogen-specific PCR primers (based on ribosomal RNA genes), a Chelex resin (for rapid DNA extraction from oyster mantle tissues), and cloned H. nelsoni rRNA plasmid DNA (for use as a capture probe). Digoxigenin was incorporated into the pathogen specific PCR products, which were captured by the coated probe in a fast hybridization reaction and then detected by ELISA. The sensitivity of PCR amplification on cloned plasmid DNA was 10 fg for detection by stained agarose gel, and increased to 0.01 fg for ELISA. Positive signals were observed in infected oysters using the PCR-ELISA technique. This method may be applicable to early detection of infection. PMID- 10373624 TI - Trout CYP1A3 Gene: Recognition of Fish DNA Motifs by Mouse Regulatory Proteins. AB - : Transcriptional up-regulation of mammalian CYP1A1 genes by dioxin is known to require binding of dioxin to the Ah receptor (AHR), subsequent interaction of this ligand-receptor complex with the AHR nuclear translocator (ARNT), and binding of this heterodimer to aromatic hydrocarbon response elements (AHREs) located in the 5' flanking sequences. From the rainbow trout (Oncorhyncus mykiss), we have isolated and sequenced the CYP1A3 gene-spanning 4.0 kb and containing seven exons and six introns-and 1897 bp of the 5' flanking region. The transcription start site was determined by primer extension analysis. Five putative AHREs were found between -451 and -1820, with an overlap of AHRE3 and AHRE4 sharing 1 bp. The 5' flanking region of the trout CYP1A3 gene was fused to the firefly luciferase (luc) reporter gene and transiently transfected into mouse hepatoma Hepa-1c1c7 wild-type (wt) cell cultures and three benzo[a]pyrene resistant mutant lines: c2, containing less than 10% levels of functional AHR; c4, defective in ARNT; and c37, deficient in CYP1A1 metabolism. We compared the trout CYP1A3 promoter-luc constructs with mouse and human CYP1A1 promoter-luc constructs. All of our trout CYP1A3 promoter data are consistent with dioxin inducible luciferase activity being controlled by two or more AHREs via cooperativity with a GC-rich region (-1852)-as has previously been demonstrated for AHREs in mammalian CYP1A1 promoters. The dependence of trout CYP1A3 promoter activity on the AHR and on the ARNT, and the enhancement of CYP1A3 promoter activity in the absence of CYP1A1 metabolic capacity, are all similar to that with mammalian CYP1A promoters. These findings indicate that the DNA motifs in trout, and the mouse liver proteins that bind to these motifs, are evolutionarily conserved elements. PMID- 10373625 TI - Molecular Species Identification of Newly Hatched Hawaiian Amphidromous Gobioid Larvae. AB - : This report describes a method for the determination of species identity of newly hatched larvae of five sympatric Hawaiian amphidromous gobioids (Lentipes concolor, Sicyopterus stimpsoni, Awaous guamensis, Stenogobius hawaiiensis, and Eleotris sandwichensis). Polymerase chain reaction (PCR) was used to amplify a homologous section of the cytochrome b (Cyt b) region of the mitochondrial genome (mtDNA) from adults of all five species. The resulting PCR-amplified DNA was subjected to restriction fragment length polymorphism (RFLP) analysis producing species-specific restriction patterns. PCR products from the five species were sequenced to substantiate correct amplification, restriction site locations, and fragment sizes. The sequence data were also used to construct a phylogenetic tree. Individual, newly hatched, wild-caught larvae of amphidromous gobioids of unknown species affinity were sorted into six morphotypes based on physical characteristics. These typed larvae and those from two species that spawned in captivity were subjected to the same molecular analysis as the adults. The RFLP results from adults and larvae were compared, allowing larval morphotypes to be assigned to the appropriate species. These comparisons permitted construction of an identification key to the newly hatched larvae of these species based solely on physical characteristics for use in future field studies.Key Words: fish larvae, species identification, Hawaii, gobioidea, PCR, RFLP.http://link.springer ny.com/link/service/journals/10126/bibs/1n2p167.html PMID- 10373626 TI - Association of Foreign DNA with Sperm of Gilthead Seabream, Sparus aurata, After Sonication, Freezing, and Dimethyl Sulfoxide Treatments. AB - : The association of foreign DNA with gilthead seabream (Sparus aurata) sperm was enhanced relative to simple coincubation by sonication, freezing, dimethyl sulfoxide and polyethylene glycol treatments. Sonication yielded the strongest dot blot signals, equivalent to 250 to 380 foreign DNA copies per spermatozoa. We are unaware of previous reports attempting to associate DNA with ultrasound for fish or elsewhere. However, no or negligible foreign DNA was evident in 1- and 2 day-old fish larvae resulting from eggs fertilized with sonicated or frozen sperm. PMID- 10373627 TI - Ribosomal Proteins S27E, P2, and L37A from Marine Invertebrates. AB - : Single complementary DNAs encoding sequences for 40S ribosomal proteins related to S27E from the American lobster Homarus americanus and mussel Mytilus galloprovincialis were characterized. Single genes for ribosomal proteins L37A and P2 from the gumboot chiton Cryptochiton stellerii are similarly described. The lobster S27E protein contains the highly conserved cysteine residues, suggesting its likely designation in the C4 protein family containing zinc finger motifs. The lobster S27E protein also appears to have an intermediate gene copy number between lower and higher euckaryotes. Expression of the S27E protein in lobster hepatopancreas was slightly elevated during several postmolt and premolt stages. Chlorinated pesticide treatment significantly reduced S27E expression in hepatopancreas, indicating that this gene is responsive to endogenous and exogenous cues. PMID- 10373628 TI - Prolidase in the Marine Sponge Suberites domuncula: Enzyme Activity, Molecular Cloning, and Phylogenetic Relationship. AB - : The enzyme prolidase hydrolyzes the peptide bond that involves the imino nitrogen of proline or hydroxyproline; hence, it catalyzes the final step in collagen degradation. From mammals it is known that this enzyme plays a major role in the recycling of proline for collagen synthesis and can be considered to be essential for the control of cell growth. The dominant organic exoskeleton in sponges, especially in Demospongiae, is collagen and the collagen-related spongin. Here we demonstrate that crude extracts of the demosponge Suberites domuncula contain prolidase or prolidase-like activity. The complementary DNA encoding the putative prolidase was cloned from a library of the same animal. Two different forms of cDNAs, termed SDPEPD1 and SDPEPD2, were identified, coding for the putative polypeptides PEPD_SD-1 with a molecular mass of 55,805 Da and PEPD_SD-2 with 51,684. Evidence is presented suggesting that the two different transcripts originate from the same gene but are formed by an alternative splicing event. We conclude that demosponges contain the activity as well as the gene for prolidase, a major enzyme involved in collagen metabolism, spicule formation, and cell motility. Phylogenetic analysis revealed that the sponge prolidase branches off first from the common ancestor of metazoan prolidases and later than the yeast prolidase; only distantly related are the bacterial enzymes. PMID- 10373629 TI - Identification and Expression Pattern of DNA Polymerase alpha Gene in a Marine Diatom, Skeletonema costatum. AB - : To investigate the potential of DNA polymerase alpha as a marker for DNA replication in phytoplankton, two gene fragments that showed a high degree of similarity with eukaryotic DNA polymerase alpha were cloned from two strains of a diatom, Skeletonema costatum (Greville) Cleve. The gene fragments amplified with the polymerase chain reaction were 397 and 396 bp in length, respectively. The deduced amino acid sequences showed 44% to 61% similarity to the corresponding regions of DNA polymerase alpha sequences of eukaryotic organisms ranging from yeast to humans. The similarity was especially high in three evolutionarily conserved regions within the amplified fragments. Further, hybridization patterns from Southern blotting confirmed that the amplified fragments were an integral part on the genome of S. costatum. In batch cultures abundant messenger of DNA polymerase alpha appeared in the late exponential phase and the early stationary phase. This pattern suggests that DNA polymerase alpha expression is associated with actively dividing cells. PMID- 10373630 TI - Isolation of Biopterin-alpha-glucoside from Spirulina (Arthrospira) platensis and Its Physiologic Function. AB - : A fluorescent substance was isolated from the cyanobacterium with a yield of 4.5 mg per 10 g of dried Spirulina (Arthrospira) platensis cells by gentle extraction and ethanol fractionation followed by column chromatography. The fluorescent substance, which has absorption maxima at 256 nm and 362 nm (pH 8.4), was identified as biopterin-alpha-glucoside by spectrophotometry and nuclear magnetic resonance spectroscopy. Biopterin-alpha-glucoside prevented decolorization of the photosynthetic pigments, chlorophyll a, phycocyanin, and carotenoids in photosynthetic vesicles of Spirulina platensis cells, by ultraviolet irradiation. PMID- 10373631 TI - Dipeptidyl peptidase IV: a cell surface peptidase involved in regulating T cell growth (review). AB - The CD26 antigen is identical with the cell surface ectopeptidase dipeptidyl peptidase IV (DP IV, EC 3.4.14.5). The post proline cleaving substrate specificity makes DP IV relatively unique among other proteases. Numerous cytokines, chemokines and other bioactive peptides are potential substrates of DP IV, but knowledge about the real in vivo substrates is still very limited. CD26 represents an accessory surface molecule playing an important role in the process of activation and proliferation of human lymphocytes. The molecular events mediated by this ectoenzyme are only partly established and the necessity of DP IV enzymatic activity for its signalling capacity has been controversial. This review out-lines evidence for an involvement of DP IV in the regulation of immune response and focuses on the putative role of the catalytic domain of this peptidase. Inhibition of the catalytic activity can provoke many cellular effects, including induction of tyrosine phosphorylations and p38 MAP kinase activation as well as suppression of DNA synthesis and reduced production of various cytokines. TGF-beta1, the production and secretion of which is increased after DP IV inhibition, supposedly mediates the observed suppressive effects by maintaining p27kip expression levels which leads to a cell cycle arrest in G1. Moreover, anti-CD3-induced signalling pathways can be strongly affected by DP IV inhibition. Thus, the enzymatic activity or at least the interaction of effectors with the catalytic domain of CD26 seem to be important for crucial functions of this cell surface antigen. PMID- 10373632 TI - Role of alanyl aminopeptidase in growth and function of human T cells (review). AB - Alanyl aminopeptidase (APN, CD13) is highly expressed in human monocytes, and anti-CD13 monoclonal antibodies are well established routine markers in leukaemia typing. Due to activation or malignant transformation other leukocyte subpopulations including human T cells exhibit significant APN-gene and surface expression. The function of leukocyte APN is poorly understood, especially the knowledge of physiological ligands/substrates of the enzyme is limited. Abnormal expression of APN on malignant lymphocytes, the activation-dependent induction of APN expression in peripheral T cells and the strong anti-proliferative effects of aminopeptidase inhibitors lead to the interesting hypothesis of a linkage of APN expression and/or function to leukocyte growth. In support of this hypothesis we detected mutations in the APN-gene of patients suffering from leukaemia or lymphoma. This review outlines evidence for APN contributing to the regulation and realisation of lymphocyte growth and function by modulating the mRNA expression of IL-2, IL-1 receptor antagonist, and TGF-beta1 and increasing the activity of MAP kinase p42/Erk2. PMID- 10373633 TI - Apoptosis induced by propolis in human hepatocellular carcinoma cell line. AB - Propolis has been reported to exhibit a wide spectrum of activities including antibiotic, antiviral, anti-inflammatory, immunostimulatory and tumor carcinostatic properties. We showed propolis induced apoptosis in a human hepatoma cell line (SNU449) by FITC-Annexin V/PI staining. We also compared the apoptosis inducing effect between Korean and Commercial (Sigma # p-1010) propolis. There was no difference on apoptosis between them. PMID- 10373634 TI - Arsenic trioxide induces apoptosis of oesophageal carcinoma in vitro. AB - The arsenic compounds in traditional Chinese medicine have been recorded to have therapeutic effects on the treatment of psoriasis, syphilis, rheumatosis and a number of malignant tumours. Recent studies showed that arsenic trioxide can induce clinical remission in patients with acute promyelocytic leukemia, including those who have relapsed after retinoic acid treatment. however, the mechanism of how arsenic trioxide targets tumour cells is not clearly understood. We have examined the effects of arsenic trioxide on oesophageal carcinoma cell line EC8712. Our results demonstrated that the growth and survival of tumour cells were markedly inhibited by arsenic trioxide. The half dose effect (ED50) was at the concentration of 1 microM. Electron microscopic study demonstrated that EC8712 tumour cells treated with arsenic trioxide display a typical morphological appearance of apoptosis, including chromatin condensation and fragmentation of the nuclei. In contrast, no apoptotic features were observed in tumour cells without arsenic trioxide treatment. TUNEL assay also showed the biological features of apoptosis in cells treated with arsenic trioxide. Flow cytometry analyses showed that apoptotic peak was identified in arsenic trioxide treated cells but not in the control. Apoptotic cells in arsenic trioxide treated group account for 35% of total cell populations after three days treatment at a dose of 3 microM. In short, our results suggested that the anticancer effect of arsenic trioxide is due, at least in part, to the induction of apoptosis in cancer cells. PMID- 10373635 TI - Tenascin expression in gastric cancer with special emphasis on the WHO-, Lauren-, and Goseki-classifications. AB - Ample evidence has been provided concerning the presence of tenascin in various histological subtypes of gastric cancer. However, conflict and discussion still persist regarding the correlation with different classification systems and prognostic impact. Therefore, we studied 203 adenocarcinomas of the stomach with special emphasis to the WHO-classification, Lauren's and Goseki's subtypes as well. The immunohistochemical ABC-method was applied using a monoclonal anti human-tenascin antibody. 30% of all tumours showed a distinct staining reaction. Tubulo-papillary carcinomas (WHO) revealed a significantly stronger reactivity than signet-ring subtypes. Adenocarcinomas of intestinal type (Lauren) were significantly more positive than the diffuse types. Mucin-poor tumours (Goseki I+III) stained positive in a much higher degree compared to mucin-rich subtypes (Goseki II+IV). However, no correlation could been demonstrated regarding TNM stage or prognosis. PMID- 10373636 TI - Human fibroblasts transfected with an ATM antisense vector respond abnormally to ionizing radiation. AB - ATM, the gene mutated in ataxia-telangiectasia (A-T), mediates multiple cellular responses to DNA damage. A-T homozygotes have a high risk of cancer and exhibit spontaneous chromosomal instability, and cultured A-T cells react abnormally to ionizing radiation. We have developed an ATM antisense vector that confers an A-T phenotype on normal cells. An episomal antisense vector was created that contained a 1.3 kb segment of the ATM cDNA, and was transfected into normal human fibroblasts. Intracellular levels of ATM protein were typically reduced 10-fold in antisense-expressing (GM639-46alpha) clones. GM639-46alpha clones exhibited the low threshold for radiation-induced apoptosis, low clonogenic survival, and cell cycle defects normally seen in A-T cells. Transfection with the corresponding ATM sense strand vector had no effect on the behavior of normal cells, and neither vector affected the behavior of A-T cells. Our results demonstrate that interference with ATM gene expression recreates the A-T phenotype in normal cells, and provide functional evidence linking the ATM gene to cellular DNA damage responses. The ATM antisense vector should prove a useful tool for studying ATM function in a variety of normal, mutant, and malignant cell lines. PMID- 10373637 TI - Neurosteroids in the cerebellar Purkinje neuron and their actions (review). AB - Although the brain is a target site of steroid hormones supplied by peripheral steroidogenic glands, it is now established that the brain itself also synthesizes steroids de novo from cholesterol in a variety of vertebrates. Such steroids synthesized in the brain are called neurosteroids. Because certain structures in vertebrate brains have the capacity to produce neurosteroids, the identification of neurosteroidogenic cells in the brain is essential to understand the physiological role of neurosteroids in brain functions. In the brain, glial cells are considered to play a major role in neurosteroid formation and metabolism. Both oligodendrocytes and astrocytes are the primary site for neurosteroidogenesis. However, the concept of neurosteroidogenesis in neurons in the brain has long been unclear. Recently, we demonstrated neurosteroidogenesis in the Purkinje cell, a typical cerebellar neuron, in mammals and other vertebrates. Pregnenolone sulfate, one of neurosteroids synthesized in the cerebellar Purkinje cell, may contribute to some important events in the cerebellum by modulating neurotransmission. Progesterone, produced as other neurosteroid in this neuron only during neonatal life, may be involved in the promotion of neuronal and glial growth and neuronal synaptic contact in the cerebellum. This review summarizes the advances made in our understanding of neurosteroids, produced in the Purkinje neuron, and their actions. PMID- 10373638 TI - Preliminary investigation of resistance of plasmid DNA to neon ion beams using transformation into recipient Escherichia coli cells. AB - The sensitivity of the vector plasmid pKmK8 DNA to neon ion beams was studied under dry conditions. This plasmid contains the cloning vector pJKKmf(-) and a 3.6 kbp segment encoding the Escherichia coli crp gene that can be used in mutagenesis analysis. The survival curve of the plasmid indicated an exponential profile and a D10 value of about 16 kGy in the E. coli wild-type strain for DNA repair capability. This was similar to the D10 value for the shuttle vector plasmid pZ189 DNA. Therefore, it was assumed that the excision repair system in E. coli was not effective for repairing DNA lesions induced by irradiation with heavy ion beams. PMID- 10373639 TI - Tumor suppressor gene p16 (CDKN2A) mutation status and promoter inactivation in head and neck cancer. AB - The p16INK4A (CDKN2A/MTS1) putative tumor suppressor gene encodes a cyclin dependent kinase inhibitor which plays an important role in the regulation of the G1/S phase cell cycle checkpoint. A high frequency of various p16 gene alterations were consequently observed in many primary tumors. P16 can be inactivated by different mechanisms: i) homozygous deletion, ii) methylation of the promoter region or iii) point mutation. In order to investigate p16 alterations in head and neck cancer (HNC) we analyzed 70 primary tumors of the larynx, pharynx and oral cavity including their corresponding normal mucosa for mutation inactivation by direct sequencing exon 2. We detected only one so far undescribed transversion G to T at position 322 (Asp108Tyr) and a known polymorphism (Ala148Thr) in five cases. The methylation status of the p16 promoter region was analyzed by an improved highly sensitive methylation-specific PCR protocol. P16 methylation inactivation was found in 16 of 55 cases (29%). Our data indicate that p16 point mutations in HNC are less frequent, but inactivation by methylation of the promoter region could be involved in genesis and progression of HNC. PMID- 10373641 TI - Matrix metalloproteinase-1 expression is a prognostic factor for patients with advanced gastric cancer. AB - Proteolytic activity of cancer cells is an important factor in metastasis. This study examined the relationship between MMP-1 expression of gastric cancer cells and peritoneal metastasis. MMP-1 expression was found in 76/103 (75.2%) cases examined and was significantly associated with both peritoneal metastasis and lymph node metastasis (p<0.05, respectively). The prognosis of patients with MMP 1 positive tumor was significantly worse than that of patients with MMP-1 negative tumor (p<0.05). These findings suggested that MMP-1 might be a prognostic factor in case of advanced gastric cancer and might be useful in determining whether or not adjuvant therapy was indicated for patients at high risk of peritoneal recurrence. PMID- 10373640 TI - Migration of mesothelioma cells correlates with histotype-specific synthesis of extracellular matrix. AB - Three human pleural malignant mesothelioma cell cultures (MM) of epithelioid (E1), fibrous (F1) and byphasic (B1) histotype were studied in their synthesis of the extracellular matrix (ECM) components laminin (LM), fibronectin (FN), type IV collagen (cIV), and in their chemotactic and haptotactic migration towards the ECM produced proteins. MM-B1 showed the highest FN synthesis and release; MM-E1 produced the highest quantity of basement membrane constituents LM and cIV; MM-F1 weakly produced and released FN, LM and cIV. MM-B1 had the highest chemotactic and haptotactic motility, MM-F1 migrated toward the lowest concentration of LM while had reduced chemotactic activity toward FN and cIV; MM-E1 had the lowest migratory activity toward each ECM substrate. We demonstrated that three MM of different histotype are characterized by different ECM production and that these differences determine a variable ability of each MM to spread and migrate towards ECM substrates. PMID- 10373642 TI - The MutT motif family of nucleotide phosphohydrolases in man and human pathogens (review). AB - Human cells express at least eight members of the MutT motif protein (or nudix hydrolase) family. These enzymes are believed to eliminate toxic nucleotide derivatives from the cell and regulate the levels of important signalling nucleotides and their metabolites. Six have been fully or partially characterized: i) hMTH1 is a nucleoside triphosphatase which restricts AT-->CG transversions by specifically degrading the oxidized nucleotide 8-oxo-dGTP; ii) hAPAH1 preferentially degrades the signalling dinucleotide Ap4A; iii) DIPP is unusual in hydrolysing two seemingly unrelated signalling substrate groups - the dinucleotides Ap6A and Ap5A, and the diphosphoinositol polyphosphates; iv) DIPP2 is closely related to DIPP; v) hYSAH1 is an NDP-sugar hydrolase which prefers ADP ribose, and vi) hGFG is a protein of unknown function encoded by the antisense transcript of the basic fibroblast growth factor gene. Although not yet associated with known hereditary or acquired disorders, the functional loss of any one of these hydrolases would be expected to be detrimental to cellular function. Furthermore, the ialA invasion gene of Bartonella bacilliformis and other invasive pathogens encodes a MutT motif Ap4A hydrolase while poxviruses express two MutT motif proteins, at least one of which is essential for infectivity. This protein family, therefore, occupies a position of some importance in controlling human health and disease. PMID- 10373643 TI - T cell regeneration in HIV-infected subjects under highly active antiretroviral therapy (review). AB - The initial idea that high amounts of cytopathic virus produced everyday can drive high CD4+ T cell production seemed logical and explained the progressive CD4+ T cell depletion observed in HIV-infected subjects. It was hypothesized that the CD4+ T lymphocyte production was increased up to 70-fold in HIV-infected subjects. Determination of the CD4+ T cell production was based on the kinetics of CD4+ T cell recovery following initiation of highly active antiretroviral therapy (HAART). However, this analysis is limited by: i) the assumption that blood CD4+ T cells are representative of the lymph node T cells; and ii) the lack of estimates of CD4+ T lymphocyte turnover in healthy HIV-negative subjects. Several immunologists have expressed caution regarding the assumptions used in modeling CD4+ T cell dynamics. Recent findings clearly show that blood is not representative of lymphoid tissues and invalidate the conclusion of high CD4 turnover drawn from blood studies on HIV-infected subjects. Indeed, when blood and lymph node compartments are considered together, we find that HIV-infected subjects naive to antiretroviral have similar or lower CD4+ T cell production, as compared to healthy subjects. This observation suggests an impaired T cell renewal capacity in HIV-1 infected patients. PMID- 10373644 TI - Usefulness of serological determinations of soluble 53 kDa protein in follow-up of melanoma patients. AB - The role of the soluble 53 kDa antigen (s53) determinations in follow-up of melanoma patient was studied. high performance liquid chromatography (HPLC) was used to measure serum levels of s53 antigen after its partial isolation on gel fiberglass (GFG) affinity chromatography columns. Two main proteins were isolated from these columns as representatives of soluble tumor-associated antigens (TAA) with molecular masses of 64 and 53 kDa. In a Western immunoblot analysis, the 53 kDa protein exhibited a strong positive reaction to the commercial p53 antibody. HPLC isolated both antigens with individual variations and significant differences in their concentrations in patients from different groups. The presence of metastases was manifested by a significant increase in the serum levels of both isolated TAA. This finding is in accordance with our previous observations on the role of the soluble 53 kDa protein in the development of colon and uterine cancers. We conclude that the determination of the serum level of the s53 antigen is of diagnostic significance in the monitoring of melanoma patients. PMID- 10373645 TI - Polypoid dysplasias in ulcerative colitis and sporadic adenomas: genetic approach to the differential diagnosis (review). AB - Development of carcinoma in cases of ulcerative colitis is initiated out of regions of dysplasia. Dysplasia may present as single or multiple flat lesions, or as polypoid dysplasia (DALM-lesion). Dysplasia in DALM lesions may not be readily differentiated from that of sporadic adenomas by light microscopy. Such distinction, however, dictates therapeutic strategies. Since colectomy is treatment of choice in DALM lesions but removal of a sporadic adenoma is sufficient therapy, several attempts have been made to find markers to differentiate the two lesions. In recent years molecular genetic studies have demonstrated different genetic alterations in the dysplasia-carcinoma event in ulcerative colitis versus sporadic colorectal carcinoma, which may in fact help to differentiate the two entities. The diagnostic tools to differentiate DALM from sporadic adenomas are briefly described and a specific emphasis is given to the molecular genetic studies which may help in defining the specific entities. PMID- 10373646 TI - Changes in the findings of dynamic MRI by preoperative CAF chemotherapy for patients with breast cancer of stage II and III: pathologic correlation. AB - Preoperative neoadjuvant chemotherapy is essential for treatment of patients with breast cancer who have a large tumor mass and/or regional lymph node involvement, in terms of both tumor shrinkage and further improvement of the survival rate. In order to safely perform breast-conservation treatment for these patients, a detailed diagnostic procedure for precisely evaluating the therapeutic response is needed. Dynamic magnetic resonance imaging (MRI) is thought to be important in the evaluation of responses to neoadjuvant therapy in patients with considerably large tumors, however, few studies have detailed the changes, as depicted by dynamic MRI, that can be expected with neo-adjuvant chemotherapy. The purpose of this study was to document the changes that occur in response to neoadjuvant chemotherapy and to correlate them with the pathological findings observed in the surgical specimen. The study was performed at Kochi Medical School Hospital from 1995 to 1998. The series consisted of 31 patients with stage II and III breast cancer. Prior to and after 1-5 courses of neoadjuvant chemotherapy, dynamic MRI examinations were performed. Eight of the time-intensity curves for the 10 grade 1a tumors flattened during neoadjuvant chemotherapy, while two remained the same. Six of the curves flattened for the 14 grade 1b tumors, 7 remained the same, and one spiked. And for the seven grade 2 tumors, two of the curves flattened and five remained the same (p=0.0340). In the five grade 1 tumors, the mean after/before normalized peak signal intensity ratio was 0.42+/-0.22. In the 18 grade 2 and 8 grade 3 tumors, the mean normalized signal intensity ratios were 0.59+/-0.28, 0.88+/-0.10, respectively (p<0.05). In the 15 tumors that showed shrinkage of the linear enhancement during neo-adjuvant chemotherapy, 10 had no remarkable intraductal spreading and 9 had a negative surgical margin. In the 16 tumors that had no shrinkage of the linear enhancement during chemotherapy, 13 had remarkable intraductal spreading and 12 had a positive surgical margin (p<0.05). It is concluded that dynamic MRI is a valuable tool for determining tumor response and predicting a positive surgical margin. Breast-conservation treatment can be performed for these patients by meticulous assessment using such detailed diagnostic procedures after local tumor control by combined chemotherapy with high dose-intensity. PMID- 10373647 TI - Inhibition of peripheral blood lymphocyte apoptosis by soluble fas ligand in patients with hepatocellular carcinoma. AB - We monitored apoptosis of peripheral blood lymphocytes (PBL) obtained from patients with various degrees of chronic viral hepatitis, including hepatocellular carcinoma (HCC), in vitro and quantitated the serum levels of soluble Fas ligand (FasL) by ELISA. There was no difference in mean percent PBL mortality. However, in HCC patients, the variance was significantly high (P<0.05), and, unexpectedly, a negative association was found between the PBL mortality and the serum soluble FasL levels (P=0.035). These results suggest the inhibitory effect of serum soluble FasL on apoptosis of PBL, which may explain the induction of immunological abnormalities with the development of HCC. PMID- 10373648 TI - Heterozygous ras mutations are preserved in serially passaged human tumor xenografts and established cell lines. AB - We examined c-K-ras gene point mutations in human tumor xenografts and established cell lines as markers of genetic stability. Our previous study demonstrated the stability of c-K-ras gene mutations in human primary neoplasms and their tumor xenografts through serial passages in mice. In this study, we established 27 human cell lines derived from various human tumor xenografts in nude mice. Point mutation of the c-K-ras gene at codon 12 was found in 29.6% (8/27) of the cell lines, as well as in 29.6% (8/27) of the xenografts. The eight ras-mutated cell lines were derived from corresponding tumor xenografts carrying the ras mutation. Heterozygous ras gene mutation was confirmed in seven of the eight ras-mutated cell lines, as well as their corresponding xenografts. The incidence, type and heterozygosity of the c-K-ras gene mutation showed no discrepancies between the original xenografts and the established cell lines. From these findings, we concluded that point mutation of the c-K-ras gene was very stable in human tumor xenografts and established cell lines derived from the xenografts. PMID- 10373649 TI - The NM23 gene and its expression in oral squamous cell carcinoma. AB - The murine nm23, a putative metastasis suppressor, has three human homologues, NM23-H1, -H2, and -H3b. Several reports have suggested a low metastatic potential for neoplasms with a high expression of NM23-H1 gene, while other studies have not shown this relationship. These apparent differences in the role of NM23 in metastasis suppression might be explained by unability to discriminate between the expression of the two genes NM23-H1 and NM23-H2. The NM23-H2 product is not related to tumor progression and metastasis suppression. Two studies on human oral squamous cell carcinoma (OSCC) have been reported, both showing the NM23 product to be a metastasis suppressor factor. However, none of these two studies distinguished NM23-H1 from NM23-H2. The aim of this study was to detect the protein expression pattern of NM23-H1 product in 24 OSCCs by immunohistochemistry in paraffin-embedded tissues using a monoclonal antibody non-cross-reactive with NM23-H2. The NM23-H1 positive group showed lower frequency of lymph node metastasis, and a better grading than the NM23-H1 negative group supporting the role of NM23-H1 as metastasis suppressor factor which may be useful for predicting tumor metastasis in OSCC. PMID- 10373650 TI - Kinetic patterns in advanced gastric cancer as related to histotype and tumor extension. AB - Kinetic patterns of advanced gastric cancers were analyzed for comparison between intestinal- and diffuse-types by using the mean values of mitotic index (MI), apoptotic index (AI), the sum of the two [i.e., the turnover index (TI)] and growth index (GI), and the values of the same parameters in the three layers (upper, intermediate, lower) in which cancers were subdivided from surface to depth. Site and extent of tumors, lymph node invasion, and p53 and PCNA expression were not different between the two histotypes; tumor cell dissociation (TCD) was higher in diffuse-type cancers. Mean MI, AI, TI, and GI were not different between the two histotypes, while MI, AI, TI, and GI were higher in the upper layer of intestinal-type cancers than in that of diffuse-type. MI and GI decreased while AI increased from upper to deeper layers in intestinal-type tumors; MI, AI, and TI increase from upper to lower layers in diffuse-type tumors. In intestinal-type cancers, but not in diffuse cases, TI and GI were higher in the T2 group than in T3. This different behavior between the two histotypes is discussed. PMID- 10373651 TI - Time until initiation of tumor growth is an effective measure of the anti angiogenic effect of TNP-470 on human glioblastoma in nude mice. AB - We examined the effect of the anti-angiogenic compound TNP-470 on early tumor growth characteristics following subcutaneous implantation of 1 mm3 tissue blocks of human glioblastoma U87, in nude mice. The mice received daily injections with TNP-470, 7 mg/kg, from one day before until either 3, 7, 11, or 15 days after inoculation. The time from inoculation until initiation of exponential tumor growth was determined along with the post-therapeutic growth delay and the initial tumor doubling time (TD) for each individual tumor (n=103) on the basis of tumor volume growth curves. We found that: i) the onset of growth of U87 xenografts was effectively inhibited by concurrent treatment with TNP-470 beyond the first three days after inoculation, ii) this effect was fully reversible upon termination of therapy, and iii) the post-therapeutic growth delay was independent of the accumulated dose. These findings demonstrate that the in vivo effect of TNP-470 on tumor growth is tumor inhibitory rather than cytotoxic. The lack of effect of the anti-angiogenic compound, TNP-470, in the early 3-day schedule is consistent with the existence of an early avascular phase which precede the angiogenesis-dependent tumor growth. PMID- 10373652 TI - Unilateral metachronous lung cancers in a patient with idiopathic pulmonary fibrosis. AB - The incidence of lung cancer in patients with idiopathic pulmonary fibrosis is much higher than that in general population. We report on a case of large cell carcinoma in association with the additional occurrence, seven months later, of an adenocarcinoma of the lung. Surgical treatment was performed for each cancer, however, the outcome was poor. The association between the two disorders is discussed. PMID- 10373653 TI - Highly aggressive behavior and poor prognosis of small cell carcinoma in the stomach: flow cytometric and immunohistochemical analysis. AB - Small cell carcinoma of the stomach has an aggressive feature, and the survival rate of the patients is poor. The purpose of this study was to determine the clinical course, and effects of histopathologic characteristics of specific tumors including DNA contents and immunohistochemical aspects in patients with small cell carcinoma of the stomach. Medical records of 8 patients who presented with small cell carcinoma of the stomach were retrospectively reviewed. Primary tumors were studied by flow cytometric analysis and immunohistochemical staining for the p53 protein, PCNA (proliferating cell nuclear antigen), factor VIII related antigen (specific for endothelial cells), VEGF (vascular endothelial growth factor) and PD-ECGF (platelet-derived endothelial cell growth factor). DNA aneuploid was observed in 4 cases. Staining for the p53 product was positive in 50% of all the cases. The average PCNA labeling rate (LR) was 71.3+/-9.9%. Positive VEGF expression was found in 7 tumors and positive PD-ECGF expression was found in all tumors. The estimated median survival was 252 days for all the patients. Liver metastases were observed in 4 of the 8 patients, however, surgery and chemotherapy have given us one long-term survivor (43 months). Higher PCNA LR of small cell carcinoma may be an unfavorable characteristic of biological behavior. Moreover, both VEGF and PD-ECGF positivity are well-characterized inducers of hepatic metastasis. PMID- 10373654 TI - Anti-telomerase therapy suppressed glioma proliferation. AB - In order to determine if telomerase activity contributes to the growth of glioma cells, we constructed an anti-telomerase vector to suppress telomerase expression in glioma cells. The human telomerase (hTR)-antisense vector showed a significant suppression effect. However it did not appear to induce cell death as a stable population of cells survived more than two months following transfection. These results provide evidence that reagents aimed at inhibiting telomerase may represent a useful strategy for suppressing the growth of glioma cells. In addition to telomerase, another mechanism would also maintain telomere length, thus supporting continuous cell proliferation. PMID- 10373655 TI - Changes of host natural killer cell activity in F344 rats during gastric carcinogenesis induced by N-methylnitrosourea. AB - To evaluate the role of NK cells during gastric carcinogenesis, especially in early stage of tumorigenesis, the changes of NK activity was examined. Rats were given N-methylnitrosourea (MNU) at a concentration of 100 ppm in their drinking water for 15 weeks. Rats were sacrificed sequentially on week 15, 18, 20, and 40 of the experimental period. Histological changes such as mild erosion, regenerative changes, focal or severe atypical lesions and invasive adenocarcinoma were observed sequentially in the pyloric region. Adenomatous hyperplasia was induced in majority of the rat stomach in MNU-treated group and incidence of adenocarcinoma was 20% in 40 weeks of MNU-treated group. There was no difference in NK activity until week 20, however, it was increased in MNU treated rats at 40 week, when compared to that of untreated control group. From week 15, the ratio of pepsinogen altered pyloric gland (PAPG) between untreated control and MNU-treated rats was progressively increased, but there was no significant increment in the number of PAPG in MNU-treated rats after 20 weeks. NK activity was increased in MNU-treated rats, when compared to that of untreated control group. These results suggest that PAPG is a relatively good marker for the evaluation of progression of gastric carcinogenesis and increased NK activity is shown, especially in early stage of gastric carcinogenesis. PMID- 10373656 TI - Atypical presentation of vertebral bone metastasis from lung cancer. AB - The authors describe a case of lung cancer in a 55-year-old man who complained of back pain. Initial isotopic bone scanning showed no abnormality, however, magnetic resonance (MRI) imaging revealed bone metastasis in thoracic vertebral bone. Even when there is no typical findings of metastasis in bone scintigraphy, MRI imaging would be useful if vertebral bone metastasis is suspected. MRI imaging is an important modality to evaluate extraosseous extension and marrow invasion of metastatic tumors. PMID- 10373657 TI - Allelic loss on the short arm of chromosome 8 in oral squamous cell carcinoma. AB - Allelic deletions on the short arm of chromosome 8 (8p) are frequent events in many different types of malignant tumors, including head and neck tumors and oropharyngeal cancers. These regions are thought to harbor tumor suppressor genes. However, there has been no detailed analysis of loss of heterozygosity (LOH) on the chromosome arm 8p in oral squamous cell carcinoma (SCC). In order to estimate details of 8p loci involved in oral SCC, 32 patients with oral SCCs were examined for the LOH state 8p by PCR-LOH assay using 14 microsatellite markers. Based on our results a detailed deletion map of 8p showed LOH in at least one of the loci tested in 62.5% (20 of 32) of patients; and microsatellite instability (MI) was observed in 50% (16 of 32). The frequent LOH on this chromosome arm was identified at D8S87 and D8S258, mapped on 8p12 and 8p22, respectively. Fisher's exact test revealed no significant statistical correlation between the incidence of LOH or MI and clinicopathological features. Our observations indicate that the short arm of chromosome 8 may play a role in the pathogenesis of oral SCC; and that the two loci identified in this study may be tumor suppressor gene loci on 8p. PMID- 10373659 TI - Paclitaxel combined with cis-platin as second-line treatment in patients with advanced non-small cell lung cancers refractory to cis-platin. AB - We combined paclitaxel with cis-platin as second-line treatment in patients with non-small cell lung cancer (NSCLC) who had previously undergone first-line therapy with cis-platin combined with cytotoxic drugs other than taxanes. The aim was to evaluate the effect of this cytotoxic combination in patients with refractory tumour to cis-platin. All 36 patients in the study population were evaluable for toxicity and 35 for response. Nine patients were stage IIIa, 15 IIIb and 12 IV. Prior treatment involved cis-platin plus vindesine and epirubicin or vinblastine and mitomycin-C; second-line treatment involved cis-platin (90 mg/m2) and paclitaxel (175 mg/m2), administered once every 3 weeks with 2-6 courses per patient. Partial response (40%) was obtained in 14 patients, 8 of whom had achieved minor response or stable disease after first-line treatment. Response duration was a minimum of 3 months. Toxicity was tolerable; only neurotoxicity was grade II in 16.7% of the patients. On the basis of our results, paclitaxel can be recommended as a very effective cytotoxic drug for NSCLC patients. PMID- 10373658 TI - Patient survival and microsatellite instability in gliomas by high-resolution fluorescent analysis. AB - Deficient repair of nucleotide mismatches in the genome is considered a major factor in tumorigenesis. Such deficiency is evidenced by alterations in dinucleotide repeats of microsatellite sequences, specifically microsatellite instability (MSI) or replication errors. We investigated the frequency of MSI in human gliomas in terms of patient outcome. Frequency of MSI was estimated by examining five loci on chromosomes 2, 5, 10, 11, and 13 in 31 gliomas using high resolution fluorescent microsatellite analysis. MSI was found at all loci in only 2 malignant gliomas (6.5%). MSI was detected at the D10S197 locus in 3 of 11 glioblastomas (27.2%) and 4 of 8 anaplastic astrocytomas (50%), while no MSI was detected in low-grade gliomas. Among patients with anaplastic astrocytoma, the 4 with MSI at D10S197 died from local recurrence less than 18 months after surgery, while 3 of the patients without MSI survived for more than 20 months. MSI at D10S197 may be a prognostic marker for patients with anaplastic astrocytomas. PMID- 10373660 TI - Conservation treatment intensified with tamoxifen and CAF chemotherapy without axillary dissection for early breast cancer patients with clinically-negative axillary nodes. AB - Axillary node dissection has been a routine part of breast cancer treatment for more than 100 years. As so few patients have been shown to have positive nodes, more consideration should be given to eliminating axillary node dissection for duct carcinoma in situ (DCIS) and T1a lesions. And for patients with a T1/2N0M0 cancer of the breast, lumpectomy alone without axillary dissection followed by radiation therapy to the intact breast and regional lymph nodes should be a reasonable treatment method that avoids arm morbidity. Between September 1989 and December 1998, we treated 79 breast cancer patients with this method intensified with tamoxifen and CAF chemotherapy. Before the start of the therapy, we performed a thorough evaluation using helical CT and doppler ultrasonography to exclude patients with significant swelling of axillary lymph nodes (more than 5 mm in short diameter). Through the end of December 1998, the mean follow-up period was 52.6 months. Up to this date, only one patient of the 79 showed local recurrence within 5 years after the start of the treatment. This patient received a second lumpectomy. She then experienced lung metastases 6 months later. She is currently receiving combined chemotherapy with docetaxel and cisplatin. The cause specific survival rate of these 79 patients maintained 100% at 6 years, and no axillary failure has been experienced so far. The cosmetic results in 50 (65.8%) of the 76 patients who were alive at the end of December 1998 were rated as excellent, 26 (34.2%) as good, and none as fair or poor. Therefore, we have concluded that this method of treatment for early breast cancer could eliminate surgical damage and allow good cosmetic results, and that survival rates with this treatment are excellent. PMID- 10373661 TI - Prognostic significance of serum LDH in Ewing's sarcoma of bone. AB - The pretreatment serum lactic dehydrogenase (SLDH) levels of 618 patients with Ewing's sarcoma of the extremities (136 metastatic at presentation and 482 localized) were analyzed to evaluate whether the enzyme level had a clinical value in predicting the course of the disease. The percentage of patients with increased SLDH was significantly higher in the metastatic group than in the group of patients with localized disease (68% vs 32%; P<0.0001). In the latter group treated with neoadjuvant chemotherapy the 5-year disease-free survival rate was significantly higher in patients with normal pretreatment SLDH than in those with high levels (65% vs 41%; P<0.0001). The time to relapse was significantly shorter for patients with elevated SLDH than in patients with normal values of the enzyme. The site of the tumor was significantly related with the stage of the disease, and for patients with localized disease, with the disease survival rate, at the multivariate analyses site of the tumor and SLDH levels were independently related with the stage of disease and with prognosis. These data demonstrate that in Ewing's sarcoma of bone pretreatment SLDH have a prognostic value and should be considered in the comparison of the results achieved with different therapies and in planning new randomized clinical therapeutic trials. PMID- 10373662 TI - Modulation of neoangiogenesis in bronchial preneoplastic lesions. AB - We have previously demonstrated that vascular count significantly increases in the preneoplastic lesions of the bronchial tree, starting from very low levels in the normal epithelium to a significantly higher number of microvessels in moderate dysplastic lesions and in situ carcinomas. Vascular endothelial growth factor (VEGF) protein expression has shown to be strictly associated with neovascularization both in human cancer and in various type of preinvasive lesions. A number of studies have demonstrated that mutant p53 is involved in the regulation of angiogenesis, and immunohistochemical detection of the p53 protein is associated with p53 gene mutations. In this study we looked for possible correlation between p53 protein detection, VEGF expression and vascular count in a series of preneoplastic and neoplastic lesions of the bronchial tree in order to investigate the angiogenic pattern and its genetic control in the early steps of bronchial cancer development. Twenty-four retrospective bronchial lesions with different grades of dysplasia and a case of normal bronchial epithelium were analysed. Surgical specimens removed from patients either confirmed, or suspect for lung carcinoma were stained immunohistochemically for CD34, VEGF, and p53. There were significant increases in microvascular density (MVD), VEGF, and p53 expression from normal bronchial epithelium through moderate dysplasia to in situ carcinoma to invasive cancer and these factors were significantly associated with moderate dysplastic lesions. A statistically significant difference was observed in MVD between hyperplastic-metaplastic, moderate dysplastic lesions and in situ carcinoma. A similar pattern was also observed for VEGF and p53 protein expression but no significant difference was observed between moderate dysplastic lesions and in situ carcinoma with regard to VEGF protein expression. The association between MVD, VEGF expression, p53 mutations and preinvasive lesions of the bronchial tree suggests that neoangiogenesis is early in non-small cell lung cancer (NSCLC) development and that p53 may have an important role in promoting angiogenesis in this human model of carcinogenesis. PMID- 10373663 TI - Prognostic significance of different biomarkers in non-small cell lung cancer. AB - The purpose of our study was to examine the prognostic significance of different biomarkers [DNA content, proliferating cell nuclear antigen labeling index (PCNA LI), p53 mutation and apoptosis], in 152 surgically resected non-small cell lung cancer (NSCLC). The ploidy was carried out by densitometry; PCNA-LI, p53 and apoptosis were determined with immunohistochemistry. The results were correlated to histology, stage and patient survival. A considerable variability of the PCNA indices, ranging from 0 to 33.5% with a mean value of 7.0%, was found. DNA evaluation showed a prevalence of aneuploid tumors (62%) with a DNA index >1. Overexpression of p53 protein and apoptotic positivity were observed in low percentages of cases (16% and 32% respectively). Only stage and PCNA-LI were found to be significant prognostic factors on multivariate analysis. PCNA was superior to stage in predicting shortened survival of patients with NSCLC. PCNA immunostaining can be applied on a routine basis in formalin-fixed, paraffin embedded samples of NSCLC to predict patient prognosis and thus to identify patients in need of additional postoperative therapies. PMID- 10373664 TI - Activity of glycogen synthase kinase-3beta is down-regulated during transient differentiation of human colon cancer HT-29 cells. AB - The increased phosphorylation and activity of protein kinase B (PKB/Akt) was found early upon butyrate treatment of HT-29 cells with a potent differentiating agent, sodium butyrate. It was accompanied by the increased phosphorylation of glycogen synthase kinase-3 (GSK-3) and the inhibition of the activity of GSK 3beta to catalyze phosphorylation of its substrate, translation initiation factor eIF2B. Phosphorylation of PKB and GSK-3 in HT-29 cells was reduced by wortmannin, the inhibitor of phosphatidylinositol-3' kinase (PI3'-kinase), which is upstream activator of PKB and GSK-3 in the intracellular signalling. Modulation of the activity and phosphorylation of these protein kinases during transient in vitro differentiation of HT-29 cells indicates that control of the PI3'-kinase/PKB dependent signalling pathway may be implicated very early in the changes of malignant phenotype of HT-29 cells. PMID- 10373665 TI - Diagnosis of malignant pleural mesothelioma by supraclavicular lymph node biopsy. AB - Extrathoracic lymph node metastases in pleural mesothelioma are rare. A case of pleural mesothelioma diagnosed by supraclavicular lymph node biopsy is described. To our knowledge, this is the first reported case of pleural mesothelioma diagnosed in this manner. PMID- 10373666 TI - Flow cytometric analysis of Ki-67 in invasive ductal carcinoma of the breast: correlation with tumor and patient characteristics. AB - Of all markers associated with cellular proliferation in breast carcinoma, Ki-67 has more often been correlated with prognosis in patients with these tumors than others. To investigate the relevance of Ki-67 determination at each phase of the cell cycle in the biological assessment of mammary carcinoma we applied bivariate Ki-67/DNA content analysis on samples from 154 resected primary lesions. Three Ki 67-derived indices including an overall and G1 and S+G2M indices were generated. These values were correlated with similar indices derived from flow cytometric DNA/RNA analysis and traditional clinicopathologic factors. The results show that overall Ki-67 indices do not correlate with flow cytometric values and clinicopathologic factors. Flow-derived Ki-67 and DNA S+G2M indices were positively correlated (p<0.0001, r=0.58). High indices for the S+G2M phase derived by both Ki-67 and DNA analysis were significantly correlated with DNA aneuploidy, high tumor grade, and negative hormonal status. We conclude that the proliferative fraction (S+G2M) by either Ki-67 or DNA analyses offers more practical and clinically relevant information in assessing the proliferative activity in mammary carcinoma. PMID- 10373667 TI - Prognostic relevance of microvessel density in colorectal tumours. AB - The importance of angiogenesis as a prognostic marker has been examined in 111 colorectal cancer patients with a minimum follow-up of 5 years. Tumour sections were immunostained with pan-endothelial antibody to CD31. Microvessels were identified and counted in 5 separate areas of highest vascularity (). Analysis of the data showed that the survival was not significantly affected by tumour site, size, grade, patients' age or gender. However, a statistically significant correlation was found between microvessel density (MVD) and survival: patients with an increased number of microvessels survived longer than those with a low number of microvessels (p=0.0007). Therefore, paradoxically, unlike other tumour types, in colorectal cancer MVD appears to be an indicator of good prognosis. The reasons that MVD correlates with good or poor prognosis are likely to vary in different tumours. For instance a frequent difficult issue in colon cancer is the presence of ulceration and adjacent severe inflammation which by itself can increase vascularity. Furthermore, overall prognosis will also depend on other factors, such as oncogenes, extracellular matrix components, adhesion molecules, growth factors, degree of apoptosis and the mode of metastatic spread. PMID- 10373668 TI - Effect of transforming growth factor-beta1 on oestrogen metabolism in MCF-7 and MDA-MB-231 breast cancer cell lines. AB - Transforming growth factor-beta (TGF-beta) has been shown to inhibit the growth of mammary epithelial cells and may play a protective role in mammary carcinogenesis. In contrast, oestrogens promote the development of breast cancer. Oestrone sulphate (E1S) is a huge reservoir of active oestrogens in the breast being converted to the weak oestrogen, oestrone (E1), by oestrone sulphatase. E1 is reversibly converted by oestradiol-17beta hydroxysteroid dehydrogenase to the potent oestrogen, oestradiol (E2). The aim of this study was to assess the effect of the TGF-beta1 isoform on growth and oestrogen metabolism in the hormone dependent MCF-7 and hormone-independent MDA-MB-231 human breast cancer cell lines. The results showed that TGF-beta1 significantly inhibited cell growth and stimulated the conversion of E1S to E1 and E1 to E2 in the MCF-7 cell line. In the MDA-MB-231 cell line TGF-beta1 significantly stimulated cell growth and inhibited the interconversions between E1 and E2. In conclusion, the growth inhibitory effect of TGF-beta1 on the MCF-7 cell line would appear to confer a protective effect in breast cancer. However, its ability to increase the amount of E2 would increase the risk of breast cancer. Which of these effects predominates in vivo remains to be explored. The growth stimulatory effect of TGF beta1 on the MDA-MB-231 cell line probably acts through a mechanism independent of the effect of TGF-beta1 on oestrogen concentrations since this cell line is hormone unresponsive. PMID- 10373669 TI - Plant tannins as inhibitors of hydroperoxide production and tumor promotion induced by ultraviolet B radiation in mouse skin in vivo. AB - The anti-oxidant and anti-tumor promotion activities of several tannins extracted from plants were examined in mouse skin treated with ultraviolet B (UVB) radiation in vivo. Hydroperoxide production was found to be maximally stimulated at a UVB dose of 200 mj/cm2, beyond which no further stimulation occurred. Treatment of mouse skin with two UVB doses of 225 mj/cm2 each, applied at a 48 h interval gradually increases the hydroperoxide (HPx)-producing activity of the epidermis, which is maximally stimulated at 4 days and returns to control levels at 15 days. The magnitude of the HPx response is found to increase with repeated UVB treatments applied at a 48 h interval and reaches a maximum level following four treatments. Of the three tannins tested (Commercial TA, Tarapod TA, and Oak TA), Tarapod TA is found to be the most effective inhibitor of UVB-stimulated HPx activity. Pretreatment with Tarapod TA inhibits, in a dose-dependent manner, this HPx response to UVB radiation. Inhibition by Tarapod TA occurs when it is applied at distant times before (-12 h) or after (+24 h) UVB radiation. When applied 20 min before UVB radiation, twice a week for 25 weeks, 8 mg of Tarapod TA inhibits the incidence and yield of papillomas promoted by UVB light in initiated skin by 34 and 70% respectively. Furthermore, when 10 mg/kg of mouse body weight of Tarapod TA was injected intraperitoneally, for a period of 25 weeks, 20 min prior to UVB treatment, it inhibited the yield of papillomas by 44%, suggesting that plant tannins when administered by various means are useful photoprotectants. PMID- 10373670 TI - Early occurrence of an adenocarcinoma after allogeneic bone marrow transplantation in a patient with AML. AB - Several reports have showed an increased risk of secondary malignancies after bone marrow transplantation (BMT), especially after total body irradiation (TBI). We report on a 39-year-old female who underwent BMT with a matched unrelated donor because of acute myeloid leukemia in second complete remission. Previously, the patient received chemotherapy for induction, consolidation, maintenance and reinduction after diagnosis of relapse. Conditioning regimen consisted of cyclophosphamide and TBI. MTX and CSA was administered for GvHD prophylaxis. Engraftment was confirmed on day 28. Within 6 months following BMT, no complication occurred. Continuous complete remission was demonstrated by repeated bone marrow smears. On day 300 the patient complained of chest pain and dyspnea. X-ray and CT-scan showed thickening of the pleura and pleural effusion. A pleuracarcinosis was diagnosed by cytologic examination of a pleural aspirate. By an open thoracotomy a disseminated inoperable disease became apparent. Diagnosis of an adenocarcinoma was confirmed by histologic examination. The patient died 2 months later due to disseminated tumour in complete remission of AML. Solid tumours are rare as secondary malignancies after BMT. Usually the neoplasmas are late events occurring more than 10 years after BMT. In this case predisposing factors such as genetic disposition, long-term smoking, intensive pretransplant chemotherapy, TBI and immunosuppression may have lead to the early secondary malignancy. PMID- 10373671 TI - High resolution mapping of chromosome 6 deletions in cervical cancer. AB - Chromosome 6 is frequently affected in different tumors. However, little information exists on chromosome 6 deletions in cervical cancer. We have studied loss of heterozygosity (LOH) and microsatellite instability (MIN) in 62 invasive squamous cell carcinomas of the cervix (CC) using 19 polymorphic microsatellite markers spanning both arms of chromosome 6 and one marker located at 5p15. We found that LOH at chromosome 6 is a common feature of cervical carcinomas: 90% (56/62) of CC had LOH at least at one locus and about 58% (36/62) had LOH on both arms of chromosome 6. The highest LOH incidence was shown in HLA region (6p21.3 6p21.1) with markers D6S273 and D6S276 in 52.7% and 45.2% of informative cases respectively. Frequent LOH on 6q was found at loci D6S311 (6q24-25. 1), D6S305 (6q26) and D6S281 (6q27-6qter) in 37.8%, 33.3% and 39.0% of informative cases respectively. There was no significant correlation observed between clinical parameters of cervical cancer (age, histologic grade, clinical stages and progression) and LOH frequency. Microsatellite instability was found in 3 out of 62 cases (4.8%) at three and more loci out of 20 tested. The study shows that several regions on 6p and 6q may harbour potential tumor-suppressor genes important for cervical cancer progression. PMID- 10373672 TI - The rising incidence of breast cancer in women and prostate cancer in men. Dietary influences: a possible preventive role for nature's sex hormone modifiers - the phytoestrogens (review). AB - The rise in breast cancer in women and prostate cancer in men in Western societies this century, stimulated a search for any possibly reversible aetiological associations common to both. Apart from more citizens living to an older age the most significant associated factor in common is the dietary practices of communities most at risk, particularly a decreasing Western dependence on plant foods with increasing dietary animal fats. Plant foods, especially legumes like soy, contain phytoestrogens (plant oestrogens). They are natural hormone modifying agents and give balance to levels of circulating hormones in both sexes. Animal fats hinder hormone modulation. They retain and slowly release unwanted stored hormones. To reverse this situation Western communities could revert to Asian type diets with high soy content but a more practical approach might be to add concentrated soy or other phytoestrogen product to a fat reduced Western diet. PMID- 10373673 TI - Metallothionein expression and zinc levels in invasive ductal breast carcinoma. AB - Metallothioneins (MTs) and zinc are both known to promote progression of tumors in human cancers. A close relationship exists between MT and zinc, as synthesis of the former has been reported to be induced by the latter. To assess the relationship between MT and zinc content in breast cancer tissues, we analysed MT expression by immunohistochemistry and tissue zinc levels by atomic absorption spectrometry (AAS) in invasive ductal breast cancers and adjacent benign breast tissues. We observed overexpression of MT in breast cancer tissues and noted that zinc content in breast cancer tissues was twice that of benign breast tissues. The nuclear fraction obtained by subcellular fractionation of breast cancer tissues was found to have a higher zinc content than the plasma membrane and cytosolic fractions. Contrary to expectations, we found a significant inverse correlation of MT expression with zinc levels in breast cancer tissues, suggesting that whilst both synergistically influence growth and survival of tumor cells, they may also have divergent roles in carcinogenesis. PMID- 10373674 TI - Analysis of Ki-67, p53 and Bcl-2 expression in the dysplasia-carcinoma sequence of Barrett's esophagus. AB - The grading of dysplasia in Barrett's esophagus has prognostic importance, however observer variation limits the reliability of simple histological analysis alone. We investigated Ki-67, p53 and Bcl-2 expression in Barrett's esophagus, in the sequence from Barrett's low-grade dysplasia to high-grade dysplasia and infiltrating adenocarcinoma. Forty-four esophagectomy specimens were utilized: 39 specimens with esophageal dysplasia and adenocarcinoma and 5 specimens with esophageal dysplasia only. This gave 83 sections (2 sections for specimens with dyplasia and carcinoma) examined from 44 patients. The sections were examined for Ki-67, p53 and Bcl-2 reactivity by immunohistochemistry. Low-grade dysplasia was present in 14 sections, high-grade dysplasia in 30 sections and carcinoma in 39 sections. Ki-67 expression occurred in 2 out of 14 (14%) sections with low-grade dysplasia, in 22 out of 30 (73%) sections with high-grade dysplasia and in 34 out of 39 (87%) sections with carcinoma (p<0.001). p53 protein expression was found in 1 of 14 (7%) sections with low-grade dysplasia, in 18 of 30 (60%) sections with high-grade dysplasia and in 33 of 39 (85%) sections with carcinoma (p<0.001). Expression of Bcl-2 was found in 11 of 14 (84%) sections with low grade dysplasia but immunoreactivity was not seen in any section with high-grade dysplasia or Barrett's carcinoma. Our results indicate that overexpression of Ki 67, Bcl-2 protein and p53 mutations can be identified as early events during neoplastic progression in Barrett's esophagus. These data support the hypothesis that, in the progression of Barrett's metaplasia to adenocarcinoma, the balance of proliferation/apoptosis plays an important role. PMID- 10373675 TI - Tissue-specific expression of the p53 tumor-suppressor gene in the intestine of transgenic mice exposed to DMH and p53 antibodies. AB - We studied the tissue-specific expression of the p53 gene in different parts of the intestine of mice treated with low doses of a carcinogen and exposed to different p53 antibodies. The human p53 promoter-CAT transgenic mice were immunized with different p53 antibodies (monoclonal - PAb 421 and DO1, and polyclonal - H-p53 and anti-soluble p53 IgG) and then exposed to low doses of dimethylhydrazine (DMH). Enzymatic CAT activity was determined in the ileum and colon 8 weeks later after the final injection of DMH. Expression of the p53 transgene in the normal ileum was twice as high as in the colon. Treatment with DMH significantly decreased the expression of the p53 transgene both in the ileum (from 18% to 100%) and in the colon (from 10% to 52%). Vaccination of mice protected at least in part such a decrease. The most effective results were found after exposure of mice to polyclonal H-p53 and to a lesser extent to anti-p53 IgG. No difference was found in the effects of antibodies on the small and large intestines. We concluded that polyclonal antibodies were more effective than monoclonal ones in protection against anti-p53 action of DMH. The observation of these effects may make it possible to explain the higher antitumor activity of polyclonal antibodies. PMID- 10373676 TI - Individual variations of total and percent free serum prostatic specific antigen: could they change the indication of prostatic biopsy? AB - The aim of this study was to analyse the individual variations of total and percent free serum prostatic specific antigen (PSA) and to evaluate whether they could change the indication for prostatic biopsy. Prostatic needle biopsy was indicated in 63 patients with serum PSA between 4.0 and 10 ng/ml. A new determination of total and free PSA was done before the biopsy procedure. The time between the determinations ranged from 29 to 59 days. The total and free serum PSA determinations were performed by a double monoclonal antibody radioimmunoassay Tandem and Tandem free PSA. The median coefficient of variation for serum PSA was 12.9 in cancer free patients and 18.8 when cancer was detected, it was 32.6 and 42.2 respectively for percent free serum PSA. A 22.8% rate of discrepancy between the determinations was found when prostatic biopsy was indicated only by percent free PSA lower than 25. Sensitivity ranged from 93.3% to 100, and reduction of unnecessary biopsies between 15.2 and 21.8%. We conclude that individual variations in total and percent free serum PSA could have clinical implications because of the possibility that it changes the indication for a prostatic biopsy. PMID- 10373677 TI - Clinical relevance of molecular markers in neuroblastoma: results from a single institution. AB - Neuroblastomas, the most common extracranial solid tumors in children, present an extremely heterogeneous behaviour that can be explained in part by their genetic abnormalities. Thirty-four patients treated at the Pediatric Oncology Unit, Hospital Vall d'Hebron from 1993 to 1997 were prospectively studied to determine the relative prognostic impact of a number of clinical and molecular factors. The factors studied were: ploidy, MYCN and 1p status, and TRK-A expression, in addition to age, stage and histology. Their impact on prognosis was analyzed. In univariate analysis, advanced stage, unfavorable histology, diploidy, MYCN amplification, and 1p deletion were identified as adverse prognostic factors; TRK A expression was associated with favorable prognosis. After multivariate analysis, only MYCN amplification proved to be an independent adverse prognostic factor (p=0.03), whereas TRK-A expression identified a subset of good-prognosis patients (p=0.003). PMID- 10373678 TI - Transplacental tumor-preventive effects of polyclonal antibodies generated against the soluble 53 kDa antigen on mammary tumorigenesis in offspring. AB - This study was designed to determine whether immunizing females with polyclonal antibodies generated against the soluble 53 kDa tumor-associated antigen (s53 TAA) has a tumor-preventive effect on their progeny and whether this effect is manifested in some biochemical characteristics. Rat females were immunized before mating with anti-s53 IgG (50 microg/rat in Freund's complete and incomplete adjuvant, three times during a month) and their 5-week-old offspring was exposed to the carcinogen (dimethylbenz(a)antracene, 10 mg/rat). Results of these experiments were studied 4 months later. Vaccination of mothers decreased the tumorigenic effects of DMBA on their offspring. Blood levels of soluble TAA were analyzed in offspring of different groups. Two TAA were isolated from the blood, with molecular masses of 64 and 53 kDa. Their concentrations differed in offspring obtained from different maternal groups. Vaccination itself resulted in a marked increase in the blood levels of TAA, not only in the mothers but also in their offspring, however, this increase was not significant in tumor-bearing animals. In offspring from non-vaccinated mothers, tumorigenesis resulted in high overexpression of s53. In offspring from vaccinated mothers, a high blood level of s53 was shown even in tumor-free animals, probably due to maternal vaccination. We conclude that maternal vaccination before pregnancy increases immunoreactivity in offspring and can reduce risk of tumors in those progeny. PMID- 10373679 TI - Microsatellite alterations in superficial and locally advanced transitional cell carcinoma of the bladder. AB - Recent studies described the existence of genetic instability associated with bladder carcinogenesis. Alterations at microsatellite loci constitute a recognized tumor marker of genome instability. A series of 21 transitional cell carcinomas of the bladder (10 superficial and 11 invasive carcinomas) was analyzed for the presence of alteration in 12 microsatellite loci, in order to detect the role of microsatellite instability in genesis and progression of human bladder cancer. Our preliminary results indicate a trend to presence of microsatellite instability (MI) in invasive and undifferentiated tumors compared to superficial and differentiated forms. Eight out of 11 T2-T4 tumors presented a number of altered microsatellite >/=2 compared to one out of 10 Ta-T1 bladder carcinomas (p=0.008). Moreover, 9 out of 15 (60%) G2-G3 tumors had significantly more unstable microsatellites than those differentiated (0 out of 6) (p=0.019). Our results provide an insight into the potential usefulness of microsatellite analysis of bladder carcinoma to better understand which neoplastic forms will evolve to invasive progression and indicate that pronounced MI may be associated with more aggressive bladder carcinomas. PMID- 10373680 TI - Soluble factors from the target organ enhance tumor cell angiogenesis: role of laminin SIKVAV sequence. AB - The ability of tumor cells to respond to microenvironmental factors present in the target organ determines in part the successful development of a metastasis. In a previous work it was demonstrated that the conditioned medium (CM) from lungs of normal mice stimulates in vitro migration, proliferation and uPA activity of cells from a murine mammary adenocarcinoma moderately metastatic to lung. This CM also enhanced local and metastatic tumor growth. Here, we show that lung CM enhanced neovascularization when inoculated together with LM3 tumor cells into the skin of syngeneic mice. A similar tumor-induced angiogenesis response was obtained when lung CM was injected systemically. Western blot analysis of lung CM revealed the presence of some laminin fragments containing the sequence SIKVAV. To determine whether those molecules were responsible for the observed angiogenic effects, the CM was depleted of the peptides containing the SIKVAV sequence. We observed that the SIKVAV-depleted lung CM lost its ability to induce an enhancement of the tumor neovascular response. Our results suggest a role for the target organ in facilitating the neovascularization of tumor cells, probably through the participation of active peptides derived from the proteolytic degradation of the basement membrane component laminin. PMID- 10373681 TI - Adriamycin-ifosfamide induction chemotherapy for extremity soft tissue sarcoma: comparison of two non-randomized protocols. AB - Chemotherapeutic cytoreduction of soft tissue sarcomas may permit less radical operation. In cases of large or multi-compartmental masses, deeply seated tumors or involvement of a neurovascular bundle, down-sizing of the mass is required before limb sparing surgery can be considered. We have applied a combination chemotherapy consisting of intravenous adriamycin and ifosfamide with intra arterial cisplatin for patients with soft tissue sarcomas of the extremity as induction treatment, and switched to an intravenous-only protocol due to toxicity and management difficulties. Adjuvant chemotherapy and radiation therapy were given after limb-sparing surgery in both regimens. Fresh tumor specimens were obtained and were examined for tumor size, surgical margins, percent of necrosis, evidence of vascular or perineural invasion, and the presence of Pgp, Ki-67, p53, PCNA and bcl-2-oncoprotein. Our results in terms of percentage of tumor necrosis were comparable and even better in favor of the second regimen [38% good histological response with intravenous (i.v.)-only versus 12.5% for combined i.v. + intra-arterial (i.a.]. The clinical and radiological responses were also better for the second (i.v. only) regimen (45%) than for the first (i.v. + i.a.) regimen (12.5%). The toxicity and the inconvenience to the patients and to the treating staff were greater in the first regimen that combined intra-arterial and intravenous infusions. In the first group the failure rate is 75% within 32 months of follow-up, while it is 33% within 12 months follow-up in the second group. The immunohistochemical markers did not correlate with disease control nor with the patient outcome. Intravenous administration of ADR-IFX induction chemotherapy was more feasible than combined i.v. ADR-IFX plus i.a. cisplatin and achieved better results. PMID- 10373682 TI - Loss of DNA-mismatch repair gene expression in oral melanomas. AB - The defect of DNA mismatch repair is postulated to be responsible for malignant transformation in many types of tumours. The main aim of this study was to evaluate the expression of DNA mismatch repair proteins in 29 cases of oral melanomas and to relate this to the ploidy status of the lesions. MLH1 expression was found in 4/29, MSH2 in 6/29, PMS2 in 2/29 and PMS1 in 0/29 cases investigated. The range of positively stained cells did not exceed 50% with MSH2, and PMS2, or 5% with MLH1. Loss of the DNA mismatch repair gene expression correlated with high aneuploidy ratio, observed in totally negative cases. PMID- 10373683 TI - Epigenetic events during the process of cell transformation induced by carcinogens (review). AB - Recent studies clearly demonstrate that several environmental carcinogens lack the ability to initially induce genetic damage. In that view, multistage chemical carcinogenesis may be processed under the control of a variety of epigenetic events in addition to genotoxic impacts. The understanding of this mechanism as reviewed in this report requires knowledge of early changes induced by carcinogens in target cells, biochemical, biological and molecular reactions closely related to both sides of the growth equation: cell proliferation and programmed death. Among several cell transformation models, the most suitable for carcinogen detection and mechanistic study is the Syrian hamster embryo (SHE) cell transformation assay. This closely mimics the multistage carcinogenesis and we can examine, in a relatively short time (8 days), the mechanisms by which genotoxic and non-genotoxic agents may increase the frequency of cell transformation as a preneoplastic end-point. The mode of action of hundred of compounds, carcinogens and non-carcinogens, has been explored so far using one stage and two-stage treatment protocols. In general, with the two-stage protocol, all carcinogens, irrespective of their genotoxic or non-genotoxic potential, give unambiguous positive results. Since perturbations of cell proliferation and death are considered essential events in the process of carcinogenesis, studies have been conducted on the dysregulation of two specific parameters, the induction of ornithine decarboxylase (ODC) an enzyme related to cell proliferation, and the apoptosis rate, when SHE cells are exposed to carcinogens. In one-stage treatment (5 h-24 h), only the promoter TPA induces ODC activity, while other carcinogens do not increase this activity. Using the two-stage exposure protocol (1 h xenobiotic/5 h TPA), all carcinogens both genotoxic and non-genotoxic, are able to stimulate ODC activity above the level obtained with TPA alone. Based on the two-stage treatment with carcinogens a close relationship can be obtained between the ODC superinduction and the increase of morphological cell transformation frequency. In cancer development, it is postulated that the inhibition of apoptosis may help altered cells to escape cell death and acquire a tumorigenic phenotype. Two-stage treatment carcinogen/TPA, effectively decreases the apoptotic rate. This is accompanied by an upregulation of the Bcl-2 oncoprotein, a well-known apoptotic inhibitor. However, treatment with a non-carcinogen phthalic anhydride, also inhibits apoptosis while it does not superinduce ODC activity. Although inhibition of apoptosis is not specific to the carcinogenic compound, both superinduction of ODC activity and inhibition of apoptosis via Bcl 2 upregulation may cooperate during the early stages of the carcinogenic process. In a long-term stage transformation assay, the rate of transformed colonies is relatively low (2-8%) bringing about the slow evolution of tumoral disease in humans and tumoral induction in rodents. This could be the consequence of the activation of various cellular repair mechanisms during the exposure time. Experimental data reported so far point out that genotoxic and non-genotoxic carcinogens, thought to be more active in the initiation or in the promotion stage, must share the same stage pathway leading to cancer development. PMID- 10373684 TI - Metal ion transporters in mammals: structure, function and pathological implications. AB - Despite the importance of metal ions in several catalytic functions, there has been, until recently, little molecular information available on the mechanisms whereby metal ions are actively taken up by mammalian cells. The classical concept for iron uptake into mammalian cells has been the endocytosis of transferrin-bound Fe3+ by the transferrin receptor. Studies with hypotransferrinaemic mice revealed that in the intestine mucosal transferrin is derived from the plasma and that its presence is not required in the intestinal lumen for dietary iron absorption. This suggests that, at least in the intestine, other non-receptor-mediated uptake systems exist. The molecular identification of metal ion transporters is of great importance, in particular since an increasing number of human diseases are thought to be related to disturbances in metal ion homeostasis, including metal ion overload and deficiency disorders (i.e. anaemia, haemochromatosis, Menkes disease, Wilson's disease), and neurodegenerative diseases (i.e. Alzheimer's, Friedreich's ataxia and Parkinson's diseases). Furthermore, susceptibilities to mycobacterial infections are caused by metal ion transporter defects. The pathological implications of disturbed metal ion homeostasis confirm the vital roles these metal ions play in the catalytic function of many enzymes, in gene regulation (zinc-finger proteins), and in free radical homeostasis. Recent insights have significantly advanced our knowledge of how metal ions are taken up or released by mammalian cells. The purpose of this review is to summarize these advances and to give an overview on the growing number of mammalian metal ion transporters. PMID- 10373685 TI - A human muscle Na+ channel mutation in the voltage sensor IV/S4 affects channel block by the pentapeptide KIFMK. AB - 1. Whole cell patch clamping of transfected HEK293 cells was used to examine the effects of a pentapeptide (KIFMK) containing the proposed inactivation particle of the Na+ channel on two mutations causing myotonia. One mutation (R1448P) is located in the voltage sensor IV/S4, and the other one (G1306E) near the postulated inactivation gate within the III-IV linker. 2. In the absence of peptide, currents of wild-type (WT) and mutant human muscle Na+ channels decayed monoexponentially with inactivation time constants that were 5-fold (R1448P) and 3-fold (G1306E) larger for the mutants. Upon intracellular application of KIFMK (0.3-1 mM) the current decay became biexponential with an additional fast decaying component that increased in amplitude with depolarization. 3. Furthermore, the peptide induced large tail currents upon repolarization, indicating that KIFMK prevents inactivation by blocking open Na+ channels. The peak of this tail current decreased only slowly with depolarizations of increasing duration. The voltage dependence of this decline indicated that the dissociation rate of the charged peptide decreased with depolarization. Increased external [Na+] ([Na+]e) antagonized block by KIFMK, consistent with a pore blocking mechanism. 4. The results are discussed with regard to a three-state model for one open, an absorbing inactivated and one blocked state with voltage dependent on- and off-rates for peptide binding. The peptide had qualitatively similar effects on WT and both mutants, indicating that the freely diffusible peptide accelerates the current decay in all three clones. However, for the R1448P mutation the affinity for KFIMK was decreased and the voltage dependence of peptide block was changed in a similar way to the voltage dependence of inactivation. These data suggest that the mutation R1448P affects the voltage dependent formation of a receptor site for both the inactivation particle and KIFMK. PMID- 10373686 TI - betagamma dimers derived from Go and Gi proteins contribute different components of adrenergic inhibition of Ca2+ channels in rat sympathetic neurones. AB - 1. Using perforated-patch recordings, we have examined the part played by endogenous G-protein subunits in the alpha2-adrenoceptor-mediated inhibition of N type Ca2+ currents in sympathetic neurones. 2. Two components of ICa inhibition by noradrenaline were recorded: a prominent, high affinity and voltage-dependent pertussis toxin (PTX)-sensitive pathway and a minor, low affinity and mostly voltage-insensitive PTX-resistant pathway. 3. PTX-sensitive inhibition was reduced by microinjection of antibodies against either GalphaoA,B or Galphai1,2. The voltage-dependent fraction of inhibition was reduced by anti-Galphao but not by anti-Galphai antibody. 4. Antisense depletion of GalphaoA led to a marked reduction of noradrenaline-induced inhibition and voltage dependence. By contrast, Galphai depletion attenuated noradrenergic modulation without affecting the voltage dependence. 5. Expression of the betagamma-binding agents beta adrenergic receptor kinase 1 (C-terminus, betaARK1C-ter) or Galphai1 with a Cys3 to Ser mutation partially prevented noradrenergic inhibition while alpha transducin abolished it. Residual inhibition was mostly voltage independent in cells expressing betaARK1C-ter but was strongly reversed by depolarization in Galphai1 Cys3Ser-expressing cells. 6. Expression of the PTX-resistant Galphai1 Cys351Ile mutant in cells treated with PTX restored alpha2-adrenoceptor inhibition. This restored inhibition was weakly reversed by depolarization. Both the degree and voltage dependence of inhibition were correlated with the level of expression of the Galphai1 Cys351Ile subunit. 7. Our findings identify betagamma dimers associated with GalphaoA and Galphai as mediators of the PTX-sensitive alpha2-adrenoceptor-mediated inhibition of N-type Ca2+ channels. Different betagamma combinations may account for the differential voltage-dependent effects of Go and Gi on ICa. PMID- 10373688 TI - Active NMDA glutamate receptors are expressed by mammalian osteoclasts. AB - 1. The N-methyl-D-aspartate (NMDA) glutamate receptor, widely distributed in the mammalian nervous system, has recently been identified in bone. In this study, we have investigated whether NMDA receptors expressed by osteoclasts have an electrophysiological activity. 2. Using the patch clamp technique two agonists of the NMDA receptor, L-glutamate (Glu) and NMDA, were shown to activate whole-cell currents recorded in isolated rabbit osteoclasts. 3. The current-voltage (I-V ) relationships of the currents induced by Glu (IGlu) and NMDA (INMDA) were studied using Mg2+-free solutions. The agonist-induced currents had a linear I-V relationship with a reversal potential near 0 mV, as expected for a voltage independent and non-selective cationic current. 4. IGlu and INMDA were sensitive to specific blockers of NMDA subtype glutamate receptors, such as magnesium ions, (5R, 10S)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a, d]cyclohepten -5,10-imine (MK 801) and 1-(1,2-diphenylethyl) piperidine (DEP). The block of IGlu and INMDA by these specific antagonists was voltage dependent, strong for negative potentials (inward current) and absent for positive potentials (outward current). 5. These results demonstrate that NMDA receptors are functional in rabbit osteoclasts, and that their electrophysiological and pharmacological properties in these cells are similar to those documented for neuronal cells. Active NMDA receptors expressed by osteoclasts may represent a new target for regulating bone resorption. PMID- 10373687 TI - [Ca2+]i determines the effects of protein kinases A and C on activity of rat renal Na+,K+-ATPase. AB - 1. It is well established that the activity of Na+,K+-ATPase (NKA) is regulated by protein kinases A (PKA) and C (PKC), but results on their effects have been conflicting. The aim of this study was to examine if this is ascribed to the intracellular concentration of Ca2+ ([Ca2+]i). 2. Rat renal NKA was stably expressed in COS cells (green monkey kidney cells). Increases in [Ca2+]i were achieved with the Ca2+ ionophore A23187 and verified by direct measurements of [Ca2+]i using fura-2 AM as an indicator. The activity of NKA was measured as ouabain-sensitive 86Rb+ uptake and the state of phosphorylation of NKA was monitored with two site-directed phosphorylation state-specific antibodies. 3. Activation of PKA with forskolin decreased NKA activity by 45.5 +/- 8.9 % at low [Ca2+]i (120 nM) and increased it by 40.5 +/- 6.4 % at high [Ca2+]i (420 nM). The change in NKA activity by forskolin correlated with the level of increase in [Ca2+]i. 4. The effect of 1-oleoyl-2-acetoyl-sn-glycerol (OAG), a specific PKC activator, on the activity of NKA was also Ca2+ dependent, being inhibitory when [Ca2+]i was low (29.3 +/- 3.6 % decrease at 120 nM Ca2+) and stimulatory when [Ca2+]i was high (36.6 +/- 10.1 % increase at 420 nM Ca2+). 5. The alpha subunit of NKA was phosphorylated under both low and high [Ca2+]i conditions upon PKA or PKC activation. PKA phosphorylates Ser943. PKC phosphorylates Ser23. 6. To see if the observed effects on NKA activity are secondary to changes in Na+ entry, we measured NKA hydrolytic activity using permeabilized membranes isolated from cells under controlled Na+ conditions. A decreased activity at low [Ca2+]i and no change in activity at high [Ca2+]i were observed following forskolin or OAG treatment. 7. Purified NKA from rat renal cortex was phosphorylated and inhibited by PKC. This phosphorylation-associated inhibition of NKA was neither affected by Ca2+ nor by calmodulin, tested alone or together. 8. We conclude that effect of PKA/PKC on NKA activity is dependent on [Ca2+]i. This Ca2+ dependence may provide an explanation for the diversity of responses of NKA to activation of either PKA or PKC. PMID- 10373689 TI - Properties of single NMDA receptor channels in human dentate gyrus granule cells. AB - 1. Cell-attached single-channel recordings of NMDA channels were carried out in human dentate gyrus granule cells acutely dissociated from slices prepared from hippocampi surgically removed for the treatment of temporal lobe epilepsy (TLE). The channels were activated by L-aspartate (250-500 nM) in the presence of saturating glycine (8 microM). 2. The main conductance was 51 +/- 3 pS. In ten of thirty granule cells, clear subconductance states were observed with a mean conductance of 42 +/- 3 pS, representing 8 +/- 2 % of the total openings. 3. The mean open times varied from cell to cell, possibly owing to differences in the epileptogenicity of the tissue of origin. The mean open time was 2.70 +/- 0.95 ms (range, 1.24-4.78 ms). In 87 % of the cells, three exponential components were required to fit the apparent open time distributions. In the remaining neurons, as in control rat granule cells, two exponentials were sufficient. Shut time distributions were fitted by five exponential components. 4. The average numbers of openings in bursts (1.74 +/- 0.09) and clusters (3.06 +/- 0.26) were similar to values obtained in rodents. The mean burst (6.66 +/- 0.9 ms), cluster (20.1 +/ 3.3 ms) and supercluster lengths (116.7 +/- 17.5 ms) were longer than those in control rat granule cells, but approached the values previously reported for TLE (kindled) rats. 5. As in rat NMDA channels, adjacent open and shut intervals appeared to be inversely related to each other, but it was only the relative areas of the three open time constants that changed with adjacent shut time intervals. 6. The long openings of human TLE NMDA channels resembled those produced by calcineurin inhibitors in control rat granule cells. Yet the calcineurin inhibitor FK-506 (500 nM) did not prolong the openings of human channels, consistent with a decreased calcineurin activity in human TLE. 7. Many properties of the human NMDA channels resemble those recorded in rat hippocampal neurons. Both have similar slope conductances, five exponential shut time distributions, complex groupings of openings, and a comparable number of openings per grouping. Other properties of human TLE NMDA channels correspond to those observed in kindling; the openings are considerably long, requiring an additional exponential component to fit their distributions, and inhibition of calcineurin is without effect in prolonging the openings. PMID- 10373690 TI - Protein kinase C mediates muscarinic block of intrinsic bursting in rat hippocampal neurons. AB - 1. Acetylcholine released from basal forebrain cholinergic fibres suppresses intrinsic bursting in cortical pyramidal cells through activation of muscarinic receptors. The signal transduction pathway mediating this action is not known. We used intracellular recordings from CA1 pyramidal cells in hippocampal slices to investigate the involvement of protein kinase C (PKC) in this cholinergic function. 2. Bath-applied carbachol (CCh; 5 microM) consistently suppressed intrinsic bursting in an atropine-sensitive (1 microM) manner. 3. Intrinsic bursting was suppressed by 4beta-phorbol 12,13-dibutyrate (PDBu; 5-10 microM), a potent PKC activator, but not by the inactive phorbol ester 4alpha-phorbol 12,13 didecanoate (PDC; 50 microM). Prior application of the PKC inhibitor 1-(5 isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride (H7; 10 microM) extracellularly or intracellularly prevented the PDBu effect. 4. Pretreatment with H7, but not with the broad-spectrum kinase inhibitor N-(2-guanidino-ethyl)-5 isoquinoline-sulfonyl hydrochloride (HA1004; 10 microM), prevented the CCh induced suppression of bursting. 5. The active component of the spike after depolarization (ADP) was reduced by CCh in an atropine-sensitive manner. This effect was mimicked by PDBu, but not by PDC. It was prevented by pretreatment with H7, but not with HA1004. 6. Blocking most K+ currents with Ca2+-free, TEA containing saline induced large TTX-sensitive plateau potentials lasting > 150 ms, driven by a persistent Na+ current. These potentials were suppressed by PDBu, but not by PDC. Pretreatment with H7 prevented the PDBu-induced suppression of the plateau potentials. 7. We conclude that cholinergic suppression of intrinsic bursting in hippocampal CA1 pyramidal cells is mediated by muscarinic activation of PKC, which down-regulates the persistent Na+ current underlying slow depolarizing potentials in these neurons. PMID- 10373691 TI - Neurotrophin-3 potentiates excitatory GABAergic synaptic transmission in cultured developing hypothalamic neurones of the rat. AB - 1. Neurotrophin-3 (NT-3) supports the survival and differentiation of neurones in the central and peripheral nervous systems through a number of mechanisms that occur in a matter of hours or days. NT-3 may also have a more rapid mode of action that influences synaptic activity in mature neurones. In the present study, the effect of NT-3 on developing GABAergic synapses was investigated in 3- to 7-day-old cultures of rat hypothalamic neurones with whole-cell patch-clamp recording. 2. NT-3 induced a substantial dose-dependent potentiation of the frequency of spontaneous postsynaptic currents (sPSCs; 160 %) in developing neurones during a period when GABA evoked inward (depolarizing) current, as determined with gramicidin-perforated patch recordings. The NT-3 effect was long lasting; continued enhancement was found > 30 min after NT-3 wash-out. NT-3 evoked a substantial 202 % increase in total GABA-mediated inward current, measured as the time-current integral. Action potential frequency was also increased by NT-3 (to 220 %). 3. The frequency of GABA-mediated miniature postsynaptic currents in developing neurones in the presence of tetrodotoxin was potentiated (to 140%) by NT-3 with no change in the mean amplitude, suggesting a presynaptic locus of the effect. 4. In striking contrast to immature neurones, when more mature neurones were studied, NT-3 did not enhance the frequency of GABA-mediated spontaneous postsynaptic currents (sPSCs), but instead evoked a slight (16%) decrease. The frequency of miniature post-synaptic currents was also slightly decreased (16%) by the NT-3, with no change in amplitude. These results were recorded during a later period of neuronal maturity when GABA would evoke outward (hyperpolarizing) currents. NT-3 had no effect on the mean amplitude of GABA-evoked postsynaptic currents in either developing or mature neurones. 5. Intracellular application of K252a, a non-selective tyrosine kinase inhibitor, did not block the NT-3 effect postsynaptically. In contrast, bath application of K252a prevented the enhancement of sPSCs by NT-3, consistent with NT-3 acting through presynaptic induction of tyrosine kinase. Decreasing extracellular calcium with BAPTA or inhibiting calcium channels with Cd2+ blocked the augmentation of sPSC frequency by NT-3, suggesting that an increase of calcium entry may be required for the facilitation of NT-3. 6. Together, our results suggest NT-3 enhances GABA release during the developmental period when GABA is depolarizing and calcium elevating, but not later when GABA is inhibitory, suggesting that one mechanism through which NT-3 may influence neuronal development is via presynaptic potentiation of GABA excitation. PMID- 10373692 TI - Muscarinic receptor modulation of GABA-mediated giant depolarizing potentials in the neonatal rat hippocampus. AB - 1. The whole-cell patch clamp technique was used to study the role of muscarinic receptors in regulating the frequency of giant depolarizing potentials (GDPs) in CA3 hippocampal neurones in slices from postnatal (P) P1-P8 rats. 2. Atropine (1 microM) reduced the frequency of GDPs by 64.2 +/- 2.9 %. The acetylcholinesterase inhibitor edrophonium (20 microM) increased the frequency of GDPs in a developmentally regulated way. This effect was antagonized by the M1 muscarinic receptor antagonist pirenzepine. 3. In the presence of edrophonium, tetanic stimulation of cholinergic fibres induced either an enhancement of GDP frequency (179 +/- 79 %) or a membrane depolarization (27 +/- 16 mV) associated with an increase in synaptic noise. These effects were prevented by atropine. 4. Application of carbachol (3 microM) produced an increase in GDP frequency that at P5-P6 was associated with a membrane depolarization and an increase in synaptic noise. These effects were prevented by atropine, pirenzepine (3 microM) and bicuculline (10 microM). 5. In the presence of pirenzepine, carbachol reduced GDP frequency by 50 +/- 4 %. Conversely, in the presence of methoctramine (3 microM), carbachol enhanced GDP frequency by 117 +/- 4 %. 6. It is concluded that endogenous acetylcholine, through the activation of M1 receptors, enhances the release of gamma-aminobutyric acid (GABA), in a developmentally regulated way. On the other hand, carbachol exerts both an up- and downregulation of GABA release through the activation of M1 and M2 receptors, respectively. PMID- 10373693 TI - Long-term potentiation of GABAergic synaptic transmission in neonatal rat hippocampus. AB - 1. The plasticity of GABAergic synapses was investigated in neonatal rat hippocampal slices obtained between postnatal days 3 and 6 using intracellular recording techniques. Ionotropic glutamate receptor antagonists were present throughout the experiments to isolate GABAA receptor-mediated postsynaptic potentials (GABAA PSPs) or currents (GABAA PSCs). 2. Repetitive depolarizing pulses (20 pulses, 0.5 s duration, at 0.1 Hz, each pulse generating 4-6 action potentials) induced a long-term potentiation in the slope and amplitude of the evoked GABAA PSPs and GABAA PSCs. 3. Long-term potentiation was prevented by intracellular injection of the calcium chelator BAPTA (50 mM), or when the voltage-dependent calcium channels blockers Ni2+ (50 microM) and nimodipine (10 microM) were bath applied. 4. Repetitive depolarizing pulses induced a persistent (over 1 h) increase in the frequency of spontaneous GABAA PSCs. 5. Repetitive depolarizing pulses induced a long-lasting increase in the frequency of miniature GABAA PSCs, without altering their amplitude or decay-time constant. 6. It is concluded that the postsynaptic activation of voltage-dependent calcium channels leads to a long-term potentiation of GABAergic synaptic transmission in neonatal rat hippocampus. This form of plasticity is expressed as an increase in the probability of GABA release or in the number of functional synapses, rather than as an upregulation of postsynaptic GABAA receptor numbers or conductance at functional synapses. PMID- 10373694 TI - Postsynaptic expression of long-term potentiation in the rat dentate gyrus demonstrated by variance-mean analysis. AB - 1. Long-term potentiation (LTP) of synaptic transmission is the putative mechanism underlying learning and memory. Despite intensive study, it remains controversial whether LTP is expressed at a pre- or postsynaptic locus. A new approach was used to investigate this question at excitatory synapses from the medial perforant path (MPP) onto granule cells in the hippocampal dentate gyrus. The variance of the evoked synaptic amplitude was plotted against mean synaptic amplitude at several different Cd2+ concentrations. The slope of the variance mean plot estimates the average amplitude of the response following the release of a single vesicle of transmitter (Qav). A presynaptic modulation should not affect Qav, but a postsynaptic modulation should alter it. 2. The variance-mean technique was tested by applying the analysis before and after three different synaptic modulations: (i) a reduction in Qav by the addition of the competitive antagonist CNQX; (ii) a reduction in the average probability of transmitter release (Pav) by the addition of baclofen; and (iii) an increase in the number of active synaptic terminals (N) by increasing the stimulus strength. CNQX reduced the average synaptic amplitude and Qav to the same extent, consistent with a postsynaptic action. In contrast, neither a change in N nor Pav altered Qav. This confirms that the variance-mean technique can distinguish between a pre- and a postsynaptic site of modulation. 3. Induction of LTP increased EPSC amplitude by 50 +/- 0.4 % (n = 5) and, in the same cells, increased Qav by 47 +/- 0.6 %. There was no significant difference between the increase in EPSC amplitude and the increase in Qav. Thus, LTP of the MPP input to dentate granule cells can be explained by an increase in the postsynaptic response to transmitter. PMID- 10373695 TI - Activation of nicotinic acetylcholine receptors patterns network activity in the rodent hippocampus. AB - 1. Intracellular and extracellular recordings from area CA3 of rat and mouse hippocampal slices revealed two distinct modes of synchronous network activity in response to continuous application of muscarinic acetylcholine receptor (mAChR) agonists. At low concentrations (e.g. 0.1-1 microM oxotremorine-M), 'burst-mode' activity comprised regular individual AMPA receptor-mediated depolarizing events, each generating several action potentials. At higher concentrations (5-50 microM), 'theta-mode' prevailed in which ordered clusters of depolarizing theta frequency oscillations occurred. 2. Whilst theta-mode activity was abolished by the mAChR antagonist atropine (5 microM), the nicotinic acetylcholine receptor (nAChR) antagonists tubocurarine (100 microM), mecamylamine (100-500 microM) and dihydro-beta-erythroidine (250 microM) converted this mode of activity to burst mode. 3. Likewise, disruption of synaptically available ACh using inhibitors of choline uptake (hemicholinium-3; 20-50 microM) or vesicular ACh transport (vesamicol; 50 microM) converted theta-mode into burst-mode activity. 4. Hippocampal slices prepared 2-3 weeks after transection of the primary cholinergic efferent pathway from the medial septum exhibited reduced vesicular ACh transporter immunoreactivity but still supported nAChR-dependent theta-mode activity suggesting that ACh released from this pathway was not critical for the activation of these receptors. 5. In summary, ACh-mediated activation of nAChRs tailors the pattern of network activity into theta-frequency depolarizing episodes as opposed to synchronized individual events at much lower frequencies. PMID- 10373696 TI - Developmental changes in low and high voltage-activated calcium currents in acutely isolated mouse vestibular neurons. AB - 1. The development of low voltage-activated (LVA) and high voltage-activated (HVA) calcium currents was studied in neurons acutely dissociated from mouse vestibular ganglia at embryonic stages (E)14, 15, 17 and birth using the whole cell patch-clamp technique. 2. LVA current was present in almost all neurons tested at stages E14 to E17, although at birth this current was restricted to a few neurons. Two populations of neurons were characterized based on the amplitude of the LVA current. In the first population, LVA current densities decreased between E17 and birth by which time this current tended to disappear in most neurons. A second population of neurons with high density LVA current appeared at E17, and in this group the mean density increased during development. 3. Among HVA currents, the dihydropyridine-sensitive L-type current remained constant between E15 and birth. Over the same period, the density of N- and Q-type currents continuously increased as shown using omega-conotoxin-GVIA (N-type), and high concentrations of omega-agatoxin-IVA (Q-type). The P-type current, sensitive to low concentrations of omega-agatoxin-IVA, transiently increased between E15 and E17, and then both current density and its proportion of the global current decreased. 4. Our results reveal large modifications in the expression of voltage dependent calcium channels during embryonic development of primary vestibular neurons. The changes in the expression of LVA current and the transient augmentation of P-type HVA current occur during a period characterized by massive neuronal growth and by the beginning of synaptogenesis. These results suggest a specific role of these currents in the ontogenesis of vestibular primary afferents. PMID- 10373697 TI - Lesion-induced plasticity in rat vestibular nucleus neurones dependent on glucocorticoid receptor activation. AB - 1. We have recently shown that neurones in the rostral region of the medial vestibular nucleus (MVN) develop a sustained increase in their intrinsic excitability within 4 h of a lesion of the vestibular receptors of the ipsilateral inner ear. This increased excitability may be important in the rapid recovery of resting activity in these neurones during 'vestibular compensation', the behavioural recovery that follows unilateral vestibular deafferentation. In this study we investigated the role of the acute stress that normally accompanies the symptoms of unilateral labyrinthectomy (UL), and in particular the role of glucocorticoid receptors (GRs), in the development of the increase in excitability in the rostral MVN cells after UL in the rat. 2. The compensatory increase in intrinsic excitability (CIE) of MVN neurones failed to occur in animals that were labyrinthectomized under urethane anaesthesia and kept at a stable level of anaesthesia for either 4 or 6 h after UL, so that they did not experience the stress normally associated with the vestibular deafferentation syndrome. In these animals, 'mimicking' the stress response by administration of the synthetic GR agonist dexamethasone at the time of UL, restored and somewhat potentiated CIE in the MVN cells. Administration of dexamethasone in itself had no effect on the intrinsic excitability of MVN cells in sham-operated animals. 3. In animals that awoke after labyrinthectomy, and which therefore experienced the full range of oculomotor and postural symptoms of UL, there was a high level of Fos-like immunoreactivity in the paraventricular nucleus of the hypothalamus over 1.5-3 h post-UL, indicating a strong activation of the stress axis. 4. The GR antagonist RU38486 administered at the time of UL abolished CIE in the rostral MVN cells, and significantly delayed behavioural recovery as indicated by the persistence of circular walking. The mineralocorticoid receptor (MR) antagonist spironolactone administered at the time of UL had no effect. 5. Vestibular compensation thus involves a novel form of 'metaplasticity' in the adult brain, in which the increase in intrinsic excitability of rostral MVN cells and the initial behavioural recovery are dependent both on the vestibular deafferentation and on the activation of glucocorticoid receptors, during the acute behavioural stress response that follows UL. These findings help elucidate the beneficial effects of neuroactive steroids on vestibular plasticity in various species including man, while the lack of such an effect in the guinea-pig may be due to the significant differences in the physiology of the stress axis in that species. PMID- 10373698 TI - Adenosinergic modulation of respiratory neurones in the neonatal rat brainstem in vitro. AB - 1. The mechanism underlying adenosinergic modulation of respiration was examined in vitro by applying the whole-cell patch-clamp technique to different types of respiration-related neurones located in the rostral ventrolateral medulla of neonatal rats (0-4 days old). 2. The adenosine A1-receptor agonist (R)-N6-(2 phenylisopropyl)-adenosine (R-PIA, 10 microM; n = 31) increased the burst distance of rhythmic C4 inspiratory discharges and decreased the duration of inspiratory discharges (control: 8.00 +/- 2.49 s and 918 +/- 273 ms; R-PIA: 12.10 +/- 5.60 s and 726 +/- 215 ms). 3. Expiratory neurones demonstrated a reversible decrease in input resistance (Rin), a depression of action potential discharges and a hyperpolarization of the membrane potential (Vm) during application of R PIA (1-10 microM). Similar responses of Rin and Vm to R-PIA were evident after synaptic activity had been blocked by 0.5 microM tetrodotoxin (TTX). 4. Some of the biphasic expiratory (biphasic E) neurones, but none of the inspiratory neurones, demonstrated changes in Rin or Vm during R-PIA application. With TTX present, R-PIA did not alter Vm or Rin in biphasic expiratory or inspiratory neurones. 5. Furthermore, R-PIA decreased the spontaneous postsynaptic activities of all neurones examined. The effects of R-PIA on respiratory activity, Rin and Vm could be reversed by the A1-receptor antagonist 8-cyclopentyl-1, 3 dipropylxanthine (DPCPX; 200 nM). 6. Our data suggest that the modulation of respiratory output induced by adenosinergic agents can be explained by (1) a general decrease in synaptic transmission between medullary respiration-related neurones mediated by presynaptic A1-receptors, and (2) an inactivation, via membrane hyperpolarization, of medullary expiratory neurones mediated by postsynaptic A1-receptors. Furthermore, our data demonstrate that inactivation of expiratory neurones does not abolish the respiratory rhythmic activity, but only modulates respiratory rhythm in vitro. PMID- 10373699 TI - The role of luminal Ca2+ in the generation of Ca2+ waves in rat ventricular myocytes. AB - 1. We used confocal Ca2+ imaging and fluo-3 to investigate the transition of localized Ca2+ releases induced by focal caffeine stimulation into propagating Ca2+ waves in isolated rat ventricular myocytes. 2. Self-sustaining Ca2+ waves could be initiated when the cellular Ca2+ load was increased by elevating the extracellular [Ca2+] ([Ca2+]o) and they could also be initiated at normal Ca2+ loads when the sensitivity of the release sites to cytosolic Ca2+ was enhanced by low doses of caffeine. When we prevented the accumulation of extra Ca2+ in the luminal compartment of the sarcoplasmic reticulum (SR) with thapsigargin, focal caffeine pulses failed to trigger self-sustaining Ca2+ waves on elevation of [Ca2+]o. Inhibition of SR Ca2+ uptake by thapsigargin in cells already preloaded with Ca2+ above normal levels did not prevent local Ca2+ elevations from triggering propagating waves. Moreover, wave velocity increased by 20 %. Tetracaine (0.75 mM) caused transient complete inhibition of both local and propagating Ca2+ signals, followed by full recovery of the responses due to increased SR Ca2+ accumulation. 3. Computer simulations using a numerical model with spatially distinct Ca2+ release sites suggested that increased amounts of releasable Ca2+ might not be sufficient to generate self-sustaining Ca2+ waves under conditions of Ca2+ overload unless the threshold of release site Ca2+ activation was set at relatively low levels (< 1.5 microM). 4. We conclude that the potentiation of SR Ca2+ release channels by luminal Ca2+ is an important factor in Ca2+ wave generation. Wave propagation does not require the translocation of Ca2+ from the spreading wave front into the SR. Instead, it relies on luminal Ca2+ sensitizing Ca2+ release channels to cytosolic Ca2+. PMID- 10373700 TI - Induction of betaine-gamma-aminobutyric acid transport activity in porcine chondrocytes exposed to hypertonicity. AB - 1. We measured the rates of uptake of selected amino acids and betaine by primary cultures of chondrocytes from porcine articular cartilage after the cells had been incubated in 'isotonic' (0.3 osmol l-1) or hypertonic (0.5 osmol l-1) media. 2. Na+-dependent uptake of methylaminoisobutyric acid increased rapidly when the cells were exposed to hypertonic conditions, reached a peak after 6-9 h, and then gradually decreased so that after 24 h it was only slightly above the control value. Conversely, (Na+ + Cl-)-dependent influx of gamma-aminobutyric acid (GABA) remained low for the first 9 h of hypertonic incubation, but then increased markedly to reach a plateau value after 24-30 h. Betaine influx also increased in cells incubated in hypertonic medium, being mainly Na+ dependent after 6 h, but (Na+ + Cl-)-dependent after 24 h. 3. This pattern indicates that exposure of the chondrocytes to hypertonicity induces first amino acid transport system A and then, as this decreases again, betaine-GABA transport activity. 4. Induction of betaine-GABA transport activity did not require continuous exposure of chondrocytes to hypertonicity; but the magnitude of the increase measured at the end of a 24 h incubation period was proportional to the length of time the cells had been exposed to hypertonicity during the 24 h. 5. Isolation and culture of chondrocytes in 0.4 osmol l-1 medium, instead of 0.3 osmol l-1, significantly increased their betaine-GABA transport activity, but not their system A activity. 6. Induction of betaine-GABA transport activity was prevented by addition of either actinomycin D or cycloheximide to the medium, but no mRNA for the betaine GABA transporter known as BGT-1 was detected by Northern blot analysis of extracts of chondrocytes. PMID- 10373701 TI - Passive water and ion transport by cotransporters. AB - 1. The rabbit Na+-glucose (SGLT1) and the human Na+-Cl--GABA (GAT1) cotransporters were expressed in Xenopus laevis oocytes, and passive Na+ and water transport were studied using electrical and optical techniques. Passive water permeabilities (Lp) of the cotransporters were determined from the changes in oocyte volume in response to osmotic gradients. The specific SGLT1 and GAT1 Lp values were obtained by measuring Lp in the presence and absence of blockers (phlorizin and SKF89976A). In the presence of the blockers, the Lp values of oocytes expressing SGLT1 and GAT1 were indistinguishable from the Lp of control oocytes. Passive Na+ transport (Na+ leak) was obtained from the blocker-sensitive Na+ currents in the absence of substrates (glucose and GABA). 2. Passive Na+ and water transport through SGLT1 were blocked by phlorizin with the same sensitivity (inhibitory constant (Ki), 3-5 microM). When Na+ was replaced with Li+, phlorizin also inhibited Li+ and water transport, but with a lower affinity (Ki, 100 microM). When Na+ was replaced by choline, which is not transported, the SGLT1 Lp was indistinguishable from that in Na+ or Li+, but in this case water transport was less sensitive to phlorizin. 3. The activation energies (Ea) for passive Na+ and water transport through SGLT1 were 21 and 5 kcal mol-1, respectively. The high Ea for Na+ transport is comparable to that of Na+-glucose cotransport and indicates that the process is dependent on conformational changes of the protein, while the low Ea for water transport is similar to that of water channels (aquaporins). 4. GAT1 also behaved as an SKF89976A-sensitive water channel. We did not observe passive Na+ transport through GAT1. 5. We conclude that passive water and Na+ transport through cotransporters depend on different mechanisms: Na+ transport occurs by a saturable uniport mechanism, and water permeation is through a low conductance water channel. In the case of SGLT1, we suggest that both the water channel and water cotransport could contribute to isotonic fluid transport across the intestinal brush border membrane. PMID- 10373702 TI - Specific distribution of sodium channels in axons of rat embryo spinal motoneurones. AB - 1. The distribution of Na+ channels and development of excitability were investigated in vitro in purified spinal motoneurones obtained from rat embryos at E14, using electrophysiological, immunocytochemical and autoradiographical methods. 2. One hour after plating the motoneurones (DIV0), only somas were present. They expressed a robust delayed rectifier K+ current (IDR) and a fast inactivating A-type K+ current (IA). The rapid neuritic outgrowth was paralleled by the emergence of a fast-activating TTX-sensitive sodium current (INa), and by an increase in both K+ currents. 3. The change in the three currents was measured daily, up to DIV8. The large increase in INa observed after DIV2 was accompanied by the onset of excitability. Spontaneous activity was observed as from DIV6. 4. The occurrence of axonal differentiation was confirmed by the fact that (i) only one neurite per motoneurone generated antidromic action potentials; and (ii) 125I alpha-scorpion toxin binding, a specific marker of Na+ channels, labelled only one neurite and the greatest density was observed in the initial segment. Na+ channels therefore selectively targeted the axon and were absent from the dendrites and somas. 5. The specific distribution of Na+ channels was detectable as soon as the neurites began to grow. When the neuritic outgrowth was blocked by nocodazole, no INa developed. 6. It was concluded that, in spinal embryonic motoneurone in cell culture, Na+ channels, the expression of which starts with neuritic differentiation, are selectively addressed to the axonal process, whereas K+ channels are present in the soma prior to the neuritic outgrowth. PMID- 10373703 TI - Relations between excitability and contractility in rat soleus muscle: role of the Na+-K+ pump and Na+/K+ gradients. AB - 1. The effects of reduced Na+/K+ gradients and Na+-K+ pump stimulation on compound action potentials (M waves) and contractile force were examined in isolated rat soleus muscles stimulated through the nerve. 2. Exposure of muscles to buffer containing 85 mM Na+ and 9 mM K+ (85 Na+/9 K+ buffer) produced a 54% decrease in M wave area and a 50 % decrease in tetanic force compared with control levels in standard buffer containing 147 mM Na+ and 4 mM K+. Subsequent stimulation of active Na+-K+ transport, using the beta2-adrenoceptor agonist salbutamol, induced a marked recovery of M wave area and tetanic force (to 98 and 87% of the control level, respectively). Similarly, stimulation of active Na+-K+ transport with insulin induced a significant recovery of M wave area and tetanic force. 3. During equilibration with 85 Na+/9 K+ buffer and after addition of salbutamol there was a close linear correlation between M wave area and tetanic force (r = 0.92, P < 0.001). Similar correlations were found in muscles where tetrodotoxin was used to reduce excitability and in muscles fatigued by 120 s of continuous stimulation at a frequency of 30 Hz. 4. These results show a close correlation between excitability and tetanic force. Furthermore, in muscles depressed by a reduction in the Na+/K+ gradients, beta-adrenergic stimulation of the Na+-K+ pump induces a recovery of excitability which can fully explain the previously demonstrated recovery of tetanic force following Na+-K+ pump stimulation. Moreover, the data indicate that loss of excitability is an important factor in fatigue induced by high-frequency (30 Hz) stimulation. PMID- 10373704 TI - Endothelial cell shrinkage increases permeability through a Ca2+-dependent pathway in single frog mesenteric microvessels. AB - 1. We tested whether calcium (Ca2+)-dependent mechanisms were essential for our previous observation that a change in the endothelial cell (EC)-extracellular matrix (ECM) attachment caused an increase in microvessel hydraulic permeability (Lp) after exposure to hypertonic solutions in single perfused mesenteric microvessels in pithed frogs (Rana pipiens). 2. In microvessels where integrin dependent EC-ECM attachments were disrupted by pretreatment with the peptide Gly Arg-Gly-Asp-Thr-Pro (GRGDTP; 0.3 mmol l-1), we measured microvessel Lp after exposure to hypertonic solutions under experimental conditions that reduced Ca2+ influx into endothelial cells. 3. High K+ solutions (59.7 and 100 mmol l-1 K+) were used to depolarize the endothelial membrane and therefore to reduce the electrochemical driving force for Ca2+ influx through conductive Ca2+ channels. These solutions abolished the increase in Lp caused by hypertonic solutions in the microvessels pretreated with GRGDTP. 4. We previously suggested that the removal of albumin from the perfusate may reduce EC-ECM attachment because hypertonic solutions increased the Lp of microvessels above that due to removal of albumin alone. This additional increase in Lp was attenuated by the 59.7 mmol l-1 K+ solution and was completely abolished by the 100 mmol l-1 K+ solution. 5. Bumetanide, an inhibitor of the Na+-K+-2Cl- co-transporter and one of the mechanisms of regulatory volume increase after exposure to hypertonic solutions in endothelial cells, did not change the response of microvessels to high K+ solutions. 6. Our findings indicate that Ca2+ entry into endothelial cells via passive conductance channels is necessary to increase microvessel Lp after exposure to hypertonic solutions in microvessels where EC-ECM attachments are disrupted. PMID- 10373705 TI - Impaired flow-dependent dilatation in distal mesenteric arteries from the spontaneously hypertensive rat. AB - 1. The aim of the study was to examine the hypothesis that flow-dependent dilatation is impaired in distal mesenteric arteries from adult spontaneously hypertensive rats (SHR) compared with normotensive Wistar-Kyoto rat (WKY) controls and to assess the role of nitric oxide (NO). 2. Arterial segments were cannulated, pressurized to 80 mmHg and allowed to develop spontaneous myogenic tone. Flow was increased incrementally in vessels from both strains and responses were also assessed before and after incubation with the NO synthase inhibitor Nomega-nitro-L-arginine methyl ester (L-NAME). Responses to flow in control vessels were also assessed before and after intraluminal perfusion with antibody complement to disrupt the endothelium. 3. At a flow rate of 5 microliter min-1, arteries from the WKY dilated significantly (22 +/- 5%, P < 0.01, n = 29) compared with the diameter at zero flow, whereas arteries from the SHR did not (4 +/- 4%, n.s., n = 16). Incubation with L-NAME had no inhibitory effect on the responses to flow in either rat strain. In control arteries, antibody-complement treatment abolished the dilatation in response to both flow and acetylcholine (ACh, 1 microM). 4. We conclude that flow-dependent dilatation is impaired in distal mesenteric arteries from adult SHR compared with WKY controls. Furthermore, flow-dependent dilatation is endothelium dependent, but L-NAME insensitive, thus excluding the NO pathway in this abnormality. Impaired flow dependent dilatation may contribute to the increased peripheral resistance in hypertension. PMID- 10373706 TI - Inward rectifier potassium conductance regulates membrane potential of canine colonic smooth muscle. AB - 1. The membrane potential of gastrointestinal smooth muscles determines the open probability of ion channels involved in rhythmic electrical activity. The role of Ba2+-sensitive K+ conductances in the maintenance of membrane potential was examined in canine proximal colon circular muscle. 2. Application of Ba2+ (1-100 microM) to strips of tunica muscularis produced depolarization of cells along the submucosal surface of the circular muscle layer. Significantly higher concentrations of Ba2+ were needed to depolarize preparations from which the submucosal and myenteric pacemaker regions were removed. 3. Elevation of extracellular [K+]o (from 5.9 to 12 mM) brought membrane potentials closer to EK (the Nernst potential for K+ ions), suggesting activation of a K+ conductance. This occurred at potentials much more negative than the activation range for delayed rectifier channels (Kv). 4. Forskolin (1 microM) caused hyperpolarization and a leftward shift in the dose-response relationship for Ba2+, suggesting that forskolin may activate a Ba2+-sensitive conductance. 5. Patch-clamp recordings from interstitial cells of Cajal (ICC) revealed the presence of a Ba2+-sensitive inward rectifier potassium conductance. Far less of this conductance was present in smooth muscle cells. 6. Kir2.1 was expressed in the circular muscle layer of the canine proximal colon, duodenum, jejunum and ileum. Kir2.1 mRNA was expressed in greater abundance along the submucosal surface of the circular muscle layer in the colon. 7. These results demonstrate that ICC express a Ba2+-sensitive conductance (possibly encoded by Kir2.1). This conductance contributes to the generation and maintenance of negative membrane potentials between slow waves. PMID- 10373707 TI - Interstitial cells of cajal generate electrical slow waves in the murine stomach. AB - 1. The gastric corpus and antrum contain interstitial cells of Cajal (ICC) within the tunica muscularis. We tested the hypothesis that ICC are involved in the generation and regeneration of electrical slow waves. 2. Normal, postnatal development of slow wave activity was characterized in tissues freshly removed from animals between birth and day 50 (D50). Slow wave amplitude and frequency increased during this period. Networks of myenteric ICC (IC-MY) were present in gastric muscles at birth and did not change significantly in appearance during the period of study as imaged by confocal immunofluorescence microscopy. 3. IC-MY networks were maintained and electrical rhythmicity developed in organ culture in a manner similar to normal postnatal development. Electrical activity was maintained for at least 48 days in culture. 4. Addition of a neutralizing antibody (ACK2) for the receptor tyrosine kinase, Kit, to the culture media caused progressive loss of Kit-immunoreactive cells. Loss of Kit-immunoreactive cells was associated with loss of slow wave activity. Most muscles became electrically quiescent after 3-4 weeks of exposure to ACK2. 5. In some muscles small clusters of Kit-immunoreactive IC-MY remained after culturing with ACK2. These muscles displayed slow wave activity but only in the immediate regions in which Kit-positive IC-MY remained. These data suggest that regions without Kit immunoreactive cells cannot generate or regenerate slow waves. 6. After loss of Kit-immunoreactive cells, the muscles could not be paced by direct electrical stimulation. Stimulation with acetylcholine also failed to elicit slow waves. The data suggest that the generation of slow waves is an exclusive property of IC-MY; smooth muscle cells may not express the ionic apparatus necessary for generation of these events. 7. We conclude that IC-MY are an essential element in the spontaneous rhythmic electrical and contractile activity of gastric muscles. This class of ICC appears to generate slow wave activity and may provide a means for regeneration of slow waves. PMID- 10373709 TI - Respiratory mechanical advantage of the canine external and internal intercostal muscles. AB - 1. The current conventional view of intercostal muscle actions is based on the theory of Hamberger (1749) and maintains that as a result of the orientation of the muscle fibres, the external intercostals have an inspiratory action on the lung and the internal interosseous intercostals have an expiratory action. This notion, however, remains unproved. 2. In the present studies, the respiratory actions of the canine external and internal intercostal muscles were evaluated by applying the Maxwell reciprocity theorem. Thus the effects of passive inflation on the changes in length of the muscles throughout the rib cage were assessed, and the distributions of muscle mass were determined. The fractional changes in muscle length during inflation were then multiplied by muscle mass and maximum active stress (3.0 kg cm-2) to evaluate the potential effects of the muscles on the lung. 3. The external intercostals in the dorsal third of the rostral interspaces were found to have a large inspiratory effect. However, this effect decreases rapidly both toward the costochondral junctions and toward the base of the rib cage. As a result, it is reversed to an expiratory effect in the most caudal interspaces. The internal intercostals in the caudal interspaces have a large expiratory effect, but this effect decreases ventrally and rostrally, such that it is reversed to an inspiratory effect in the most rostral interspaces. 4. These observations indicate that the canine external and internal intercostal muscles do not have distinct inspiratory and expiratory actions as conventionally thought. Therefore, their effects on the lung during breathing will be determined by the topographic distribution of neural drive. PMID- 10373708 TI - In vitro recordings of afferent fibres with receptive fields in the serosa, muscle and mucosa of rat colon. AB - 1. Colonic afferent fibres were recorded using a novel in vitro preparation. Fibres with endings in the colonic mucosa are described, along with those in muscle and serosa, and their responses to a range of mechanical and chemical luminal stimuli. 2. Mechanical stimuli were applied to the tissue, which included stretch, blunt probing of the mucosa and stroking of the mucosa with von Frey hairs (10-1000 mg). Chemical stimuli were applied into a ring that was placed over the mechanoreceptive field of the fibre; these were distilled water, 154 and 308 mM NaCl, 100 microM capsaicin, 50 mM HCl, and undiluted and 50% ferret bile. 3. Recordings were made from 52 fibres, 12 of which showed characteristics of having endings in the mucosa. Mucosal afferents were sensitive to a 10 mg von Frey hair and were generally chemosensitive to >= 1 chemical stimulus. 4. Ten fibres showed characteristics of having receptive fields in the muscular layer. These fibres responded readily to circumferential stretch, as well as to blunt probing. 5. Twenty-seven fibres showed characteristics of having endings in the serosal layer. They adapted rapidly to circumferential stretch and responded to blunt probing of the serosa. Fifteen of 19 serosal fibres tested also responded to luminal chemicals. 6. Three fibres were unresponsive to all mechanical stimuli but were recruited by chemical stimuli. 7. This is the first characterization of colonic afferent fibres using an in vitro method and the first documentation of afferent fibres with their endings in the mucosa of the colon. These fibres are likely to be important in aspects of colonic sensation and reflex control. PMID- 10373710 TI - Spatial distribution of external and internal intercostal activity in dogs. AB - 1. The observation that the external and internal interosseous intercostal muscles in the dog show marked regional differences in mechanical advantage has prompted us to re-examine the topographic distribution of electrical activity among these muscles during spontaneous breathing. 2. Inspiratory activity was recorded only from the areas of the external intercostals with an inspiratory mechanical advantage, and expiratory activity was recorded only from the areas of the internal intercostals with an expiratory mechanical advantage. The expiratory discharges previously recorded from the caudal external intercostals and the inspiratory discharges recorded from the rostral internal intercostals were probably due to cross-contamination. 3. Activity in each muscle area was also quantified relative to the activity measured during tetanic, supramaximal nerve stimulation (maximal activity). External intercostal inspiratory activity was consistently greater in the areas with a greater inspiratory advantage (i.e. the dorsal aspect of the rostral segments) than in the areas with a smaller inspiratory advantage, and internal intercostal expiratory activity was invariably greatest in the areas with the greatest expiratory advantage (i.e. the dorsal aspect of the caudal segments). 4. This topographic distribution of neural drive confers to the external intercostal muscles an inspiratory action on the lung during breathing and to the internal interosseous intercostals an expiratory action. PMID- 10373711 TI - Changes in excitability indices of cutaneous afferents produced by ischaemia in human subjects. AB - 1. The present study was undertaken to determine whether mechanisms other than membrane depolarization contribute to the changes in excitability of cutaneous afferents of the median nerve under ischaemic conditions. 2. In six healthy subjects, axonal excitability was measured as the reciprocal of the threshold for a compound sensory action potential (CSAP) of 50% maximal amplitude. Refractoriness and supernormality were measured as threshold changes 2 and 7 ms, respectively, after supramaximal conditioning stimuli. The strength-duration time constant (tauSD) was calculated from the thresholds for unconditioned CSAPs using test stimuli of 0.1 and 1.0 ms duration. Changes in these indices were measured when subthreshold polarizing currents lasting 10 or 100 ms were applied, before, during and after ischaemia for 13 min. 3. At rest, the change in supernormality produced by polarizing currents was greater with the longer polarizing current, indicating that it took up to 100 ms to charge the internodal capacitance. 4. Refractoriness and its dependence on excitability increased more than expected during ischaemia. Supernormality was abolished during ischaemia, and reached a maximum after ischaemia but was then barely altered by polarizing current. tauSD had a similar relationship to excitability before, during and after ischaemia. 5. By contrast, during continuous depolarizing current for 8 min to mimic the depolarization produced by ischaemia, the relationship between excitability and refractoriness was the same during the depolarization as before it. 6. It is suggested that the large increase in refractoriness during ischaemia might be due to interference with the recovery from inactivation of transient sodium channels by an intra-axonal substrate of ischaemia. The post-ischaemic increase in supernormality and the lack of change with changes in axonal excitability can be explained by blockage of voltage-dependent potassium channels. PMID- 10373712 TI - Clopidogrel with aspirin is the optimal antiplatelet regimen for intracoronary stenting. PMID- 10373713 TI - New thrombolytic strategy: bolus administration of tPA and urokinase-fibrinogen conjugate. AB - Increased efficacy of thrombolytic therapy requires a comprehensive search for new and novel therapeutic strategies. Many new modified forms of plasminogen activators have been obtained by means of chemical and biological synthesis. However, clinical findings demonstrate that the reperfusion level achieved during thrombolysis remains the same for various thrombolytic agents, irrespective of an extensive search for an "ideal" thrombolytic. Thrombolytic therapy may be complicated by treatment delays, cumbersome schemes of preparation and administration, and hemorrhagic and rethrombotic events. These limitations may be overcome, at least in part, by applying combined thrombolysis with plasminogen activators exhibiting complementary actions and different pharmacokinetic profiles. The combined action of native thrombolytics allows the use of lower doses and simplified schemes of administration, yielding encouraging results in experimental models. Long-acting forms of plasminogen activators are being developed and tested in combination with tissue-type plasminogen activator as a trigger of thrombolysis. The combination of short- and long-acting plasminogen activators appears promising and potentially eligible for bolus administration to patients. On the basis of our own experimental results and data in the literature, we suggest a new thrombolytic strategy connected with the single injection of a combination of complementary and pharmacokinetically different plasminogen activators. PMID- 10373715 TI - Low molecular weight heparins in the cardiac catheterization laboratory. PMID- 10373714 TI - Hemostatic effects of 1 mg daily warfarin on post CABG patients. Post CABG Studies Investigators. AB - Although coronary bypass graft surgery has increased the survival and quality of life of many individuals, patients remain at risk of restenosis and thrombotic occlusion of the coronary arteries and bypass grafts. In the screening period for participation in the multicenter Post Coronary Artery Bypass Graft (Post CABG) trial, the effects of 1 mg daily warfarin were evaluated using paired patient samples collected prior to and after at least 21 days of treatment. In stable patients (n = 40; 39 males 1 female; 51-74 years old) who previously had undergone coronary artery revascularization (1-10 years), no alterations in prothrombin time, international normalized ratio (INR), prothrombin fragment 1.2, or the hemostatic risk factors factor VII antigen and coagulant activity, von Willebrand's factor, fibrinogen, tPA, or PAI-1 were associated with the 1 mg daily warfarin treatment. The observations reported here supported the Post CABG Studies Steering Committee decision to treat patients with 1-4 mg warfarin daily adjusted to achieve INRs not to exceed 2. 0 consistent with low-intensity therapy. PMID- 10373717 TI - Use of glycoprotein IIb/IIIa inhibition plus fibrinolysis in acute myocardial infarction. AB - Pharmacological reperfusion therapy for acute myocardial infarction with intravenous fibrinolytic agents improves survival yet fails to achieve early and complete coronary blood flow in nearly half of treated patients. In principle, glycoprotein (GP) IIb/IIIa inhibitors, potent antiplatelet agents, might improve the efficacy and clinical outcomes associated with fibrinolysis. Preclinical research suggests more rapid and effective reperfusion with combined platelet GP IIb/IIIa inhibition and fibrinolysis. Early clinical studies confirm improved early patency and more rapid electrocardiographic resolution, but increased bleeding complications, with the addition of GP IIb/IIIa antagonists to conventional fibrinolysis. Future studies may combine reduced-dose fibrinolytic therapy with GP IIb/IIIa inhibition to optimize efficacy and safety. PMID- 10373716 TI - Aspirin and ticlopidine after routine coronary stenting: the gold standard as of 1999. PMID- 10373719 TI - 67th Annual meeting of the Swiss Society of Internal Medicine and annual Swiss Medical Societies meetings. Bale, Switzerland, 15-17 April 1999. Abstracts. PMID- 10373718 TI - Emerging treatment of acute coronary syndromes with platelet glycoprotein IIB/IIIA inhibitors. PMID- 10373720 TI - The conceptualization of health. AB - The meaning of health is complex and subject to change. In this article, four conceptual models of health are presented to summarize the current meanings for health. The medical model is the most widely used definition in the United States, but the World Health Organization model has gained in popularity during the past several decades. In addition, there are other newer models--the wellness model and the environmental model--that are adding new meanings to the definition of health. By understanding and combining these different meanings, the prospects for improving medical outcomes and the quality of care are enhanced. This conceptual work is a prelude to improving health status assessment in a variety of contexts. PMID- 10373721 TI - Medicare spending by beneficiaries with various types of supplemental insurance. AB - The authors analyzed Medicare spending by elderly noninstitutionalized Medicare beneficiaries with and without supplemental insurance such as Medigap, employer sponsored plans, and Medicaid. Use of a detailed survey of Medicare beneficiaries and their Medicare health insurance claims enabled the authors to control for health status, chronic conditions, functional limitations, and other factors that explain spending variations across supplemental insurance categories. The authors found that supplemental insurance was associated with a higher probability and level of Medicare spending, particularly for Part B services. Beneficiaries with both Medigap and employer plans had the highest levels of spending ceteris paribus, suggesting a possible moral hazard effect of insurance. Findings from this study are discussed in the context of the overall financing of health care for the elderly. PMID- 10373722 TI - Strategic groups and performance in the nursing home industry: a reexamination. AB - This study further examines the relationship between strategic group membership and performance in the nursing home industry. The results indicate seven strategic groups in the industry with significant between-group differences in operating margin, average profit per patient day, catheterization rate, health deficiencies, life and safety deficiencies, and efficiency in the provision of services. The authors did not, however, find significant between-group differences in pressure sore rate, the use of restraints, or in the percentage of patients with significant unplanned weight changes. The group with the highest private-pay utilization combined with high Medicare utilization generally performed the best along all indicators. Results also suggest that strategic group membership and rural/urban location have a greater impact on performance than do ownership, chain membership, or possession of a dedicated specialty care unit. PMID- 10373723 TI - Survival and treatment for colorectal cancer Medicare patients in two group/staff health maintenance organizations and the fee-for-service setting. AB - The current study compares treatment use and long-term survival in colorectal cancer patients between Medicare beneficiaries enrolled in two large prepaid group/staff health maintenance organizations (HMOs) and the fee-for-service (FFS) setting. The study is based on 15,352 colorectal cancer cases diagnosed between 1985 and 1992 and followed through 1995. Survival differences between the HMO and FFS cases were assessed using Cox regression. Treatment differences were evaluated using logistic regression. HMO cases had a lower overall mortality than did FFS cases but not a significantly lower colorectal cancer-specific mortality. Use of surgical resection was similar between HMO and FFS cases. However, rectal cancer cases in the HMOs were more likely to receive postsurgical radiation therapy than FFS cases. Superior overall survival in the HMOs may be the result of increased colorectal cancer screening, greater use of adjuvant therapies, and selection of healthier individuals. PMID- 10373724 TI - Covering uninsured children and their parents: estimated costs and number of newly insured. AB - The Child Health Insurance Program (CHIP) supplies $20.4 billion over 5 years and nearly $50 billion over 10 years to extend health insurance to uninsured children with family incomes up to 200 percent of poverty. This article analyzes the March 1997 Current Population Survey, estimating the number of children likely to be eligible for CHIP or currently eligible for Medicaid. Of the 8.6 million parents of uninsured children, four out of five were uninsured at the time of the survey. Expanding coverage to parents as well as children could make program participation more attractive and simplify the enrollment process. If 75 percent of uninsured parents of CHIP eligible children participated, 1.7 million parents could be insured, costing federal and state governments $4 billion. Another 3.4 million parents would be insured by expanding Medicaid to cover uninsured parents of Medicaid-eligible children. PMID- 10373725 TI - Hospital conversions from for-profit to nonprofit status: the other side of the story. PMID- 10373726 TI - Non-invasive ventilation--mechanisms of benefit. AB - Over the last few years there has been growing interest in the use of non invasive ventilation (NIV) in the management of ventilatory failure in acute on chronic ventilatory failure and in stable hypercapnic patients. To understand why assisted ventilation should be effective in improving gas exchange even during spontaneous breathing a basic understanding of the pathophysiology of ventilatory failure is required and this is discussed. The etiology of ventilatory failure is likely to be multifactorial with different factors assuming greater or lesser degrees of importance even in patients with the same condition. Indeed in an individual patient there may be differences at various stages of the illness. The same is true for the mechanism of benefit from NIV. With the current state of knowledge during NIV the aim should be to rest the respiratory muscles, control nocturnal hypoventilation and improve sleep quality. In some individuals severe symptoms, as a consequence of disturbed sleep, may occur and be symptomatically improved by non-invasive ventilation during sleep. PMID- 10373727 TI - [Daytime mechanical ventilation in chronic ventilatory insufficiency]. AB - BACKGROUND: Nocturnal hypoventilation is associated with chronic ventilatory insufficiency (CVI). Noninvasive mechanical ventilation (NIV) performed overnight relieves symptoms of hypoventilation and improves daytime blood gases in CVI. In order to test whether the efficacy of NIV depends on its being applied during sleep we conducted a prospective case controlled study comparing daytime mechanical ventilation (DV) in awake patients with nocturnal mechanical ventilation (NV) given in equal quantities. PATIENTS AND METHODS: We enrolled 34 clinically stable patients (age: 56.1 +/- 12.1 years, 20 female) with CVI due to both restrictive lung and chest wall disorders and neuromuscular disease. Using a prospective case-control design, matched subjects were allocated alternately to DV and NV. RESULTS AND CONCLUSIONS: There were no significant differences between the groups in the improvement of the measured parameters; (e.g. PaCO2; DV: from 57.6 +/- 4.3 to 44.0 +/- 4.6 mm Hg, NV: from 55.5 +/- 3.5 to 45.0 +/- 4.1 mm Hg, p < 0.0001). We conclude that in many respects, when compared to NV, DV in awake patients is equally effective for the treatment of CVI. PMID- 10373728 TI - [Sleep-phase-related home therapy in congenital central hypoventilation syndrome (CCHS)]. AB - PATIENTS AND METHOD: Eight children with congenital central hypoventilation syndrome (CCHS) (aged 3 to 16 years) underwent repeated polysomnographic recordings (sleep-EEG, induction plethysmography, PtcO2, PtcCO2, PACO2, FO2, SaO2, ECG) during spontaneous breathing and during therapy. The result led to individual therapeutic plans. RESULT: During NREM sleep a close relationship between increasing EEG-delta-activity and increasing PCO2 could be observed (PCO2 max. 107 mm Hg in NREM IV). A similar effect was seen during mechanical ventilation with decreasing spontaneous respiratory activity during increasing sleep depth (PCO2 max. 89 mm Hg in NREM IV). Associated with NREM I/II and REM sleep strong variations in spontaneous breathing with consecutive variations of blood gases were observed. Hyperventilation during REM sleep (PCO2 min. 20 mm Hg) could occur with continuous mechanical ventilation. A continuous blood gas monitoring improved home therapy since blood gas adapted control of mechanical ventilation was possible now. This caused a stabilization of blood gases in sleep. CONCLUSION: Patients with CCHS show a vigilance-dependent, enlarged variability of blood gases which should be considered in the management of home therapy. Continuous monitoring and blood gas adapted mechanical ventilation obtain a stabilization of acid-base balance during sleep. Preliminary data suggest a positive effect on sleep-wake quality and mental performance. PMID- 10373730 TI - [Acceptance and long-term results of home mechanical ventilation in various thoracic diseases]. AB - BACKGROUND: Home mechanical ventilation (HMV) is an important therapy for patients with respiratory insufficiency on the basis of neuromuscular diseases (NMD), chest wall deformities (CWD) and chronic obstructive pulmonary disease (COPD). PATIENTS AND METHODS: We retrospectively analyzed the long-term results of all 144 patients (CWD = 47, COPD = 54, NMD = 43) who underwent a trial of non invasive HMV from March 1990 to September 1997. RESULTS: Twenty-eight patients did not accept the HMV (19%), 7 with CWD (15%), 17 with COPD (32%) and 4 with NMD (9%). Thirty-nine of 113 patients, who accepted HMV, completed nasal ventilation for a minimum of 1 year. For all 3 groups the hypercapnia improved significantly (CWD 58 +/- 6 to 48 +/- 4 mm Hg, p < 0.001, COPD 61 +/- 7 to 46 +/- 6 mm Hg, p < 0.001, NMD 53 +/- 8 to 42 +/- 6 mm Hg, p < 0.001). CONCLUSION: HMV improves the hypercapnic ventilatory failure independent of the underlying disease. The rate of acceptance is lower in patients with COPD in comparison to CWD and NMD. PMID- 10373729 TI - [Invasive and non-invasive home ventilation--changes between 1982 and 1996]. AB - PATIENTS AND METHODS: In our centre, 111 patients with chronic ventilatory insufficiency (33 females, 78 males, age 48 +/- 18 years, range 3 to 76 years) were treated by intermittent positive pressure ventilation between 1982 and 1996. Underlying diseases were neuromuscular diseases in 29%, sleep-related hypoventilation in 26%, kyphoscoliosis in 15%, chronic obstructive airway disease in 15%, and post-tuberculosis syndromes in 12%. Singular indications were 1 bronchiectasis, 1 lung fibrosis and 1 cystic fibrosis. RESULTS: Until 1991, most patients were ventilated via tracheostoma (10 of 16), in the following years 87 of 95 patients could be ventilated via a nasal or facial mask. Ventilation mode was a controlled one in 80 patients and an assisted one in 31 patients, average ventilation time during night was 6 to 8 hours. In the majority of patients hypercapnia was not only removed during ventilation but also at daytime as an indicator of improvement of ventilatory insufficiency accomplished by a clearly better quality of life and daytime activity. Ten patients (9%) died due to their underlying diseases, 5 of them in the first year of intermittent ventilation. PMID- 10373731 TI - [Exercise tolerance of patients under nasal intermittent positive pressure ventilation (nIPPV)]. AB - BACKGROUND: There are only a few papers concerning with exercise tolerance of patients under nasal intermittent positive pressure ventilation (nIPPV). PATIENTS AND METHOD: Therefore since 1996 we routinely checked exercise tolerance of our nIPPV-patients when admitted to the hospital. Till March 1997 we had carried out 1386-minute walking tests (6-min WT) in 111 patients. QUESTIONS: Is there an improvement of exercise tolerance in the course of nIPPV-therapy? Are hypoxemia or hypercapnia occurring during exercise-test? METHODS: The 6-min WT was performed after one practice walk. The patients got oxygen in case of a preexisting oxygen therapy or in case of an oxygen saturation below 85% before starting. Blood gas analyses were carried out before and after stopping the test. Oxygen saturation and heart rate were registered continuously. The distance walked was measured. Twenty-one patients were tested before introducing nIPPV therapy and 3 months after home mechanical ventilation (HMV). RESULTS: The average distance walked amounted only 283 +/- 82 m (norm in healthy persons: 800 m). pO2 decreased from 69 +/- 11 to 58 +/- 12 mm Hg, pCO2 increased from 47 +/- 8 to 49 +/- 8 mm Hg. Oxygen saturation (SaO2) fell from 92 +/- 5 to 80 +/- 10%, heart rate increased from 104 +/- 18 to 130 +/- 23 beats/min. The distance walked changed not significantly from 282 +/- 109 to 308 +/- 71 m. Six patients could be tested a 3rd time after 6 months HMV. The distance walked was 315 +/- 103 m (also no significant difference). CONCLUSIONS: Everyday activity can cause severe hypoxemia in nIPPV patients. Ambulatory oxygen therapy should be considered in each case. A significant improvement of exercise tolerance under nIPPV therapy is not yet proven. Our data only show a tendency towards an increase. PMID- 10373732 TI - [Pharmacological stress echocardiography--a new non-invasive follow-up examination of intermittent ventilation]. AB - BACKGROUND: Patients with chronic respiratory insufficiency frequently develop pulmonary hypertension. Non-invasive intermittent ventilation is usually very successful in these patients to improve blood gas exchange and clinical symptoms. Alterations of pulmonary hemodynamics during non-invasive intermittent ventilation are rarely described. Pharmacological stress echocardiography of the right heart is a new method to examine pulmonary hemodynamics. Aim of this study was to answer the question whether non-invasive intermittent ventilation improves pulmonary hemodynamics. PATIENTS AND METHOD: Five patients are examined prior to and during non-invasive intermittent ventilation by right ventricular stress echocardiography at rest and during exercise. Pulmonary arterial pressure was registered and compared. The effectiveness of intermittent ventilation was evaluated with respect to blood gas analytic values. RESULTS: During non-invasive intermittent ventilation all 5 patients improved their pulmonary arterial mean (PAP mean) and systolic pressure, but no statistical significant changes could be observed during the 4.5-months follow-up. CONCLUSION: Non-invasive intermittent ventilation improves the pulmonary hemodynamics at rest and during exercise the results not being significant. PMID- 10373733 TI - The art of interface. Tools for administering noninvasive ventilation. AB - BACKGROUND: Since the choice of interface plays a large role in the effectiveness of noninvasive ventilation, there is a need for information about what is available, how to make a selection, what to do when a problem occurs, and how to provide proper care and monitoring. MATERIAL AND METHODS: Here we will present some of the different types of interfaces available, different problems presented by each, and guidelines for making choices among them. PMID- 10373734 TI - [Long-term breathing via tracheostoma]. AB - PATIENTS AND METHODS: From 1988 to 2/1997 we had introduced intermittent positive pressure ventilation (IPPV) in 298 patients. In most cases non-invasive nasal mask ventilation was possible, in 21 patients (7%) a tracheostoma was necessary. These 21 patients were analysed retrospectively due to age, sex, diagnose, ventilation mode, course of illness, home care and costs. RESULTS: We had 13 male and 8 female patients, aged 49 years on average (min. 2, max. 84). 90% had neuromuscular diseases especially muscle dystrophies. Ventilation therapy was performed volume controlled with the cannula unblocked during daytime and blocked at night. Eighteen patients had industrial cannulas (72% Shiley, 28% Rusch), 3 patients used silver cannulas. Daily ventilation amounted 24 hours in 7 patients, 6 to 14 hours in 14 patients. During the observed time 7 patients remained in stable health situation, in 9 patients the underlying disease was progressive and 5 of them died. IPPV was performed 50.7 months on an average, in living patients 68.8 months, in died 7.6 months. Fifteen patients lived at home, 5 were cared in nursing home, 1 patient stayed in hospital. Outside the hospital the bigger part of costs was paid by sick funds and care funds, the smaller part by social welfare offices. Often costs were divided. Total costs for caring about 24 hours ventilated patient at home amounted up to 21,000 German marks each month. PMID- 10373735 TI - [Dysphagia due to tracheotomy]. AB - FUNDAMENTALS: Dysphagia is a common appearance after tracheostomy especially in patients with neurologic diseases of the following reasons: 1. fixation of trachea and larynx to the scin leads to lack of laryngeal elevation in a forward and upward direction, 2. pressure in the upper esophagus may be increased following high cuff pressure, 3. reduction of hypopharyngeal and laryngeal sensitivity and alteration of the coughing mechanism, 4. loss of olfactorial and gustatorial stimulation of the swallowing process. CONCLUSION: Consequently noninvasive ventilation modes are recommended alternatively in patients with respiratory insufficiency. PMID- 10373736 TI - [Endotracheal complications after long-term ventilation. Noninvasive ventilation in chronic thoracic diseases as an alternative to tracheostomy]. AB - PATIENTS AND METHODS: In this present retrospective study we examined 62 long term ventilated patients, whose weaning from respirator failed, for endoscopic airway complications and the frequency of consecutive surgery required. Furthermore noninvasive volume-controlled intermittent ventilation was evaluated as an alternative method to tracheostomy for maintaining mechanical ventilation and weaning of patients with chest wall disorders, neuromuscular and chronic obstructive lung disease. RESULTS: 25 patients with endotracheal tube and 37 with tracheostomy who had been long-term ventilated in different intensive care units for 18 +/- 12 respectively 57 +/- 27 days (19 +/- 12 days via endotracheal tube) could be weaned successfully consequently using a volume-controlled intermittent ventilation via an individually adapted face mask. We found 2 patients of the group with endotracheal intubation (median age 59 +/- 15 years, 11 female, 14 male, median duration of mechanical ventilation via tube 18 +/- 12 days) to have visible injuries of the respiratory tract without consecutive surgery being necessary. All of them were successfully weaned from respirator via noninvasive ventilation (in 2 of them completely spontaneous breathing was re-established, 23 patients needed intermittent ventilation at home). Of the 37 patients with tracheostomy (median age 59 +/- 15 years, 15 female, 22 male, median duration of mechanical ventilation 57 +/- 27 days, tracheostomy on day 19 +/- 12) 19 cases (51%) showed endoscopically visible injuries of the respiratory tract of whom 7 cases (19%) were severe and made consecutive surgery necessary. 29 patients were discharged with noninvasive ventilation at home, 5 needed further invasive ventilation via tracheostomy and 3 patients breathed spontaneously without ventilatory support. The incidence of severe tracheal stenosis following long term ventilation via tracheostomy was nearly 20% (1 tracheoesophageal fistula) and needed surgical treatment. CONCLUSION: As even duration of ventilation via tracheal tube and mode of ventilation before transfer to our clinic was comparable in both groups noninvasive ventilation is an appropriate alternative to tracheostomy following endotracheal intubation for maintaining ventilatory support, especially for patients with chronic ventilatory insufficiency. PMID- 10373737 TI - [Percutaneous dilatational tracheostomy]. AB - BACKGROUND: Tracheostomy provides a method for long-term ventilation in intensive care, which reduces the risk of necrotizing lesions of the pharyngeal and laryngeal mucosa. Since the introduction of the percutaneous dilatational tracheostomy, experienced physicians are able to perform bedside tracheostomies. This presentation reviews the complication rate and long-term outcome of percutaneous dilatational tracheostomy. PATIENTS AND METHOD: The method was applied in 57 patients following previous orotracheal intubation averaging 7.8 days (3 to 15 days). Underlying diseases were sepsis/SIRS in 29, stroke in 7, cerebral hypoxemia after cardiopulmonary resuscitation in 10, trauma in 7, prolonged weaning in 2, primary neurological diseases in 2. RESULTS: The following complications occurred during the procedure: 1 major and 7 minor bleedings. 2 subcutaneous emphysemas, 1 mediastinal emphysema following tracheal injury. No complication required surgical intervention. In the follow-up 17 patients (30%) died from their underlying disease, none from complications of the tracheostomy. After removed of the tracheal tube, in 39 patients the stoma closed spontaneously within 7 to 14 days. In 8 patients the tracheostoma persisted for more than 3 months, but no clinically relevant tracheal stenosis was found. CONCLUSION: Percutaneous dilatational tracheostomy is a safe procedure easy to perform in intensive care units. Bronchoscopic control is necessary to avoid complications. PMID- 10373738 TI - [Continuous flow apnoeic ventilation via intratracheal oxygen insufflation]. AB - BACKGROUND: In patients with disturbed gas-exchange (e.g. COPD) intratracheal oxygen insufflation (ITO2) improves oxygenation and reduces the minute ventilation. We use a bronchoscopic technique of intratracheal catheter placement in unintubated patients. In a patient with a pink-puffer emphysema after endoscopical insertion of the catheter ITO2 induced a "continuous flow apnoeic ventilation" (CFAV). CASE REPORT: A patient (female, 58 years) with a pink-puffer emphysema was admitted to the ICU with acute on chronic respiratory failure due to acute laryngitis. Because of laryngitis associated upper airway obstruction a non-invasive mechanical ventilation could not be performed. The ensuing high flow ITO2 (10 l/min) induced a CFAV characterized by no chest wall movement and adequate ventilation as reflected by stable, elevated PaCO2 (between 118 and 125 mm Hg), which could be maintained for 4 hours. After an ensuing short-term invasive mechanical ventilation and the administration of high dose glucocorticoids the patient was successfully extubated and the clinical status improved continuously. CONCLUSION: In a patient with an acute on chronic respiratory failure due to end-stage emphysema ITO2 induced CFAV and stabilized the clinical status. Especially in patients with end-stage emphysema, who are likely to be difficult to be weaned from the respirator ITO2 may be a feasible technique in order to bridge an emergency situation. PMID- 10373739 TI - [Non-invasive positive pressure ventilation in cardiogenic pulmonary edema]. AB - PATIENTS AND METHOD: 30 patients being admitted to our intensive care unit with severe cardiogenic pulmonary edema received non-invasive positive pressure ventilation (NIPPV) via face mask. RESULT: 29 responded well, 1 patient had to be intubated. Within 30 minutes those who responded well showed a significant improvement of the following parameters: rise of peripheral saturation from 75.5 to 90.1% and of pH from 7.24 to 7.29, decrease of pCO2 from 60.7 to 48.8 mm Hg and of systolic blood pressure from 144 to 124 mm Hg. Mean duration of ventilation was 6 h 55 min. Mean stay in intensive care unit was 2 days. No patient required ventilator support within 24 hours after NIPPV. Four patients died during hospital stay as a result of their underlying disease but not due to pulmonary edema. Observed side effects were vomiting in 4 cases during NIPPV without aspiration and 3 cases of skin lesions which healed uneventfully. CONCLUSION: Key role for this highly effective method seems to be the rapid improvement of left ventricular function during NIPPV. PMID- 10373740 TI - [Brain syndrome and home ventilation--diagnosis, therapy and consequences]. AB - BACKGROUND: A growing number of patients with neuromuscular diseases has been treated with mechanical ventilatory support during the last years. In some of these patients acute or chronic organic brain syndromes complicate the situation. PATIENTS AND METHODS: We present case reports of 5 patients who have been ventilated at home with neuromuscular diseases but who also suffered from brain syndromes of different etiology. CASE REPORTS: (1) Multifactorial acute organic brain syndrome after intensive care therapy. (2) Recurrent paranoid psychosis correlating with hypercapnia. (3) Oneiroids (awake dreamings). (4) Refusal of the rejection of mechanical ventilation facing dyspnea. (5) Persistent vegetative state after polyradiculomyeloencephalitis. DISCUSSION: Patients with hypoventilation or mechanical ventilation often present with various types of brain syndromes. Careful history taking and the use of laboratory and imaging techniques allow the differentiation in primary and secondary (metabolic) brain syndromes. With respect to the fact that home ventilation has marked consequences for the patient and his carer organic disorders of brain function raise two questions: (1) Is the patient able to understand and critically reflect the consequences of long-term mechanical ventilation? (2) Is the patient able to understand and manage the practical aspects and skills of home-ventilation. e.g., does he have the mental capacity for a sufficient compliance? From our point of view patients with severe brain syndromes should not be selected for home ventilation. PMID- 10373741 TI - [From tracheostomy to non-invasive mask ventilation: a study in children with congenital central hypoventilation syndrome]. AB - BACKGROUND: Children with congenital central hypoventilation syndrome (CCHS) have to be ventilated during sleep due to respiratory insensitivity to CO2. This long term mechanical ventilation sometimes requires a tracheostomy during infancy, leading to increased risk of infections and of tracheal problems, and later on to stigmatization and restrictions in social life. PATIENTS AND METHOD: We therefore evaluated non-invasive mask ventilation in 4 children between 6 and 15 years of age, who had been ventilated via tracheal canula since early infancy under polysomnographic control. RESULTS: Best results were obtained with standard face masks in connection with pressure controlled timed ventilation. In 1 child we used a volume-controlled ventilator. The lack of dyspnea in these patients can worsen the acceptance of a face mask, which is more uncomfortable than a tracheal cannula. In 2 children we waited with the definite closure of the tracheostomy due to pavor-like symptoms and laryngeal closure during sleep and problems in acceptance of the mask, respectively. In the other 2 children we could demonstrate effective non-invasive mask ventilation during temporary tracheal closure for several nights. Therefore the tracheostomy was definitely closed. Long-term follow-up with home monitoring showed effectiveness of non-invasive ventilation in these cases. PMID- 10373742 TI - [Importance of blood transfusion in anemic patients with COPD and unsuccessful weaning from respirator]. AB - BACKGROUND: Imbalance of load-capacity relationship in severe COPD may lead to ventilatory failure. Additionally, in such patients, anemia may aggravate the ventilatory load. In this retrospective study we investigated whether anemia patients undergoing long-term ventilation benefit from transfusion. CASE REPORTS: We studied 5 anemic patients (hemoglobin: 8.7 +/- 0.8 g/dl) with COPD in whom trials of weaning from the ventilator had been unsuccessful. After transferral to our regional weaning centre, blood was transfused to increase the hemoglobin value to approximately 12 g/dl or higher. Subsequently, all patients could be successfully weaned within a short period. CONCLUSION: We conclude that in difficult to wean anemic patients blood transfusion should be considered and may lead to successful weaning. PMID- 10373743 TI - [Nasal mechanical ventilation in children]. AB - BACKGROUND: Nasal mechanical ventilation is not only applicable to adults but also in childhood when necessary. PATIENTS AND METHODS: Thirty-six children suffering from various neuromuscular diseases were brought up by their parents to learn nasal mechanical ventilation. Thirty children had to be ventilated, because of symptomatic ventilatory failure, reduced ventilatory muscle capacity or hypercapnia. RESULTS: Thirty-five children could be adapted to nasal mechanical ventilation, 1 girl needed a naso-oral mask. All children wanted to continue with ventilation because they realized the benefit. Their symptoms disappeared. CONCLUSIONS: The management of ventilatory failure should be the same in adults and children. Nasal mechanical ventilation is indeed a good possibility for children even in early childhood. Children should be introduced to mechanical ventilation at the beginning of the symptoms of ventilatory failure. PMID- 10373744 TI - [Concept of apartment-sharing groups for children and adolescents treated with long-term ventilation]. AB - Children on long-term ventilation without adequate support have already been a topic of reports a few times. An inquiry conducted by the Vestische Kinderklinik confirmed this for 14 patients on ventilation in the area of Westfalen-Lippe. According to estimates in the weekly magazine "Der Spiegel" there are about 200 affected children and adolescents. Even in situations of continuous pressure, basic human needs and rights are not to be neglected, they are to be taken into consideration at each medical, therapeutic and pedagogic intervention. From such a "holistic" point of view the living surroundings of those patients should not be exclusively or mainly defined by ICU needs. Even more, due to the development in intensive care medicine, there should be the possibility of a compromise under "regular living circumstances", ensuring an optimal cooperation of medical "life security" as well as psychological (including the family) and pedagogical guidance and help. With the concept of "flat sharing groups for ventilated children and adolescents" we promote a complex model that seems to be able to fulfil the requirements stated above. The idea of integration as well as the paradigm of "self-determined life" should be taken into account. PMID- 10373745 TI - [The role of non-invasive positive pressure ventilation in lung volume reduction surgery of pulmonary emphysema--a survey of German hospitals]. AB - BACKGROUND: Since the first publication by Cooper et. al. in 1994 of lung volume reduction surgery (LVRS) of emphysema a marked respiratory failure with hyperkapnia (PaCO2 > 55 Torr) has been regarded as an exclusion criterion for LVRS. PATIENTS AND METHOD: In a survey in German hospitals the question was asked whether non-invasive nasal ventilation (NIPPV) has a role in the management of LVRS. Of 12 hospitals 6 had experience with NIPPV and LVRS in a total of 19 patients with a mean FEV1 of 0.64 +/- 0.101. RESULTS: LVRS improved FEV1 by 0.20 +/- 0.181. Preoperative NIPPV was short (< 6 months) in 8 patients and resulted in improvement of physical condition and getting the patient used to NIPPV for better perioperative management. In 5 cases NIPPV was used on a long-term basis in order to allow the patient to be included in the LVRS program. In fact 7 of these 13 patients needed ventilation perioperatively, and 4 had to continue long term NIPPV after surgery. In further 3 patients NIPPV was applied only perioperatively. One patient had to resume NIPPV after 15 months. Two patients started NIPPV 1 resp. 12 months after surgery. Two patients had bronchial cancer which was resected. Four patients died: 1 perioperatively after intubation, 2 after 3 resp. 13 months due to respiratory failure, 1 for cancer relapse after 20 months. CONCLUSION: NIPPV may be helpful in the planning and management of LVRS in patients with ventilatory failure with hypercarbia. PMID- 10373746 TI - [Intermittent positive pressure ventilation after sternectomy]. AB - BACKGROUND: The indication for intermittent positive pressure ventilation (IPPV) is the symptomatic hypercapnic ventilatory insufficiency. Beside the improvement of life quality and extension of life time the aim of IPPV is a reduction of the secondary effects of chronic hypoventilation in order to stabilize the symptoms. PATIENTS AND METHODS: We examined 2 patients after sternectomy because of osteomyelitis who developed a symptomatic ventilatory insufficiency together with recurrent dys- and atelectasis and pneumonia, resistant against to antibiotic treatment. After initiation of IPPV the patients turned to a clinically stable condition. The nocturnal oxygen saturation improved as well as the daytime blood gas analysis. In these patients the indication for IPPV was not only the symptomatic hypercapnic ventilatory insufficiency but also the prophylaxis of recurrent dys- and atelectasis and pneumonia. Antibiotic therapy after sternectomy is often not successful, therefore in case of recurrent infections in patients with unstable thorax the early initiation of IPPV seems to be useful. PMID- 10373747 TI - [Diaphragm pacing in chronic hypoventilation]. AB - BACKGROUND: Diaphragm pacing in patients with intact phrenic nerve and diaphragm can be used as an alternative to mechanical ventilation. Indications cover diseases caused by central hypoventilation like C2-quadriplegia and Ondine's syndrome. Advantages are physiological ventilation with negative pressure and an improvement in articulation. CASE REPORT: We report our experiences with a patient suffering from chronic hypoventilation caused by a ventilatory pump failure following tetraparesis due to congenital toxoplasmosis. PMID- 10373748 TI - [Evaluation of daily activity in patients with COPD]. AB - BACKGROUND: Recently we could show that the daily activity in patients with hypercapnic respiratory failure as judged by the total amount of movements per day increased by 120% after 3 months of non-invasive mechanical ventilation. This study was designed to evaluate the reproducibility of the result of a movement detector in measuring daily activity of patients with COPD. PATIENTS AND METHODS: 25 outpatients (11 females, 56 +/- 12 years old) with stable non-hypercapnic COPD (FEV1 = 47 +/- 9% predicted) were examined twice during a 7 days lasting interval, one month apart using a pedometer. RESULTS AND CONCLUSIONS: The repeatability of the activity counts in the non-hypercapnic COPD patients was high (1st study period: 3,781 +/- 2,320 movements/d, 2nd study period: 3,626 +/- 2,149 movements/d) and in these patients activity correlated significantly with FEV1 (r = 0.54, p = 0.006). PMID- 10373749 TI - [Different approaches to long-term O2 therapy (LTOT) in patients using intermittent positive pressure ventilation]. AB - BACKGROUND: Until now there is no conclusion about a distinguished long-term O2 therapy in patients using IPPV. PATIENTS AND METHODS: 24 patients (6 women, 18 men, mean age 60 years, mean length of IPPV 15.6 months, mean length of long-term O2 therapy 16.7 months) with IPPV and long-term O2 therapy were studied by the topical and the past dose and the length of long-term O2 therapy. RESULTS: 1. Thirteen patients had the highest need for oxygen under strain with same need under IPPV and daytime. 2. The distribution among the others was different, 3 patients showed an elevated need for oxygen from IPPV to day-time to activity, 3 needed oxygen only under strain. 3. Changing the length was necessary in 4 patients. 4. The individual dose was changed in the course in 18 patients, reduced in 4, raised in 6 and both in 8 patients. CONCLUSION: 1. A distinguished long-term O2 therapy with testing the need in rest, under IPPV and in activity is convenient. 2. Regular controls are necessary because of the individual changings. PMID- 10373750 TI - [Quality of life of various patient groups during home mechanical ventilation]. AB - BACKGROUND: During home mechanical ventilation quality of life depends on improvement of ventilation and progress of the underlying disease. PATIENTS AND METHODS: Patients with chronic obstructive pulmonary disease (COPD; n = 11), neuromuscular diseases (NMD; n = 8) and scoliosis (n = 8) answered before and after 306 +/- 232 (64 to 910) days home mechanical ventilation a standardized and validated questionnaire (SF 36, Medical Outcomes Trust, Boston, USA). RESULTS: For all patients together physical functioning (16 +/- 22 to 24 +/- 26%), general health (33 +/- 15 to 41 +/- 20%) and vitality (25 +/- 16 to 45 +/- 22%) improved significantly (p < 0.05). For COPD vitality (22 +/- 15 to 46 +/- 22%), for patients with NMD mental health (62 +/- 9 to 70 +/- 10%) and for patients with scoliosis vitality (35 +/- 15 to 59 +/- 22%) and mental health (61 +/- 11 to 74 +/- 4%) improved. CONCLUSION: Home mechanical ventilation improves quality of life, but the improvement depends on the underlying disease. PMID- 10373751 TI - [Amyotrophic lateral sclerosis and nasal mechanical ventilation]. AB - BACKGROUND: Patients suffering from amyotrophic lateral sclerosis (ALS) can profit from nasal mechanical ventilation and improve in the quality of life. PATIENTS AND METHODS: Thirty-eight patients were introduced to nasal mechanical ventilation, using pressure- and volume-cycled respirators. The daily periods of mechanical ventilation varied from 8 to 24 hours. RESULTS: Twelve women and 26 men with ALS mostly with severe symptoms were adapted to the intermittent nasal ventilation. 80% had bulbar symptoms. Nineteen patients died till now. Even when complications occurred it was possible to use the noninvasive ventilation. CONCLUSIONS: Noninvasive mechanical ventilation is possible in spite of complications and improves the quality of life in ALS. PMID- 10373752 TI - [Effect of acupuncture on quality of life, mouth occlusion pressures and lung function in COPD]. AB - BACKGROUND: There are few data concerning the effect of acupuncture in patients suffering from COPD. We conducted a prospective randomized and placebo-controlled pilot study to analyze the effect of acupuncture according to the rules of traditional chinese medicine on quality of life, pulmonary function testing and inspiratory mouth occlusion pressures (MOP). PATIENTS AND METHODS: We randomised 10 patients with stable COPD to a 2-week treatment of 7 verum acupuncture or placebo acupuncture sessions. Before and after treatment we performed pulmonary function tests as well as an interview with the chronic respiratory disease questionnaire. MOP were taken on day 1, 5 and 7 before and after punction to assess function of the respiratory pump. RESULTS: Patients receiving verum acupuncture improved significantly with FEV1 and RV/TLC. There was an improvement of large magnitude in quality of life and a trend of lower demand of the respiratory pump. In the placebo group we saw only a slight improvement of quality of life, a deterioration of lung function parameters and a trend of higher demand of the respiratory pump. CONCLUSION: Although the small number of subjects allows no further conclusions this pilot study proves feasibility of acupuncture in COPD and shows that acupuncture is worthy for further investigation in patients suffering from COPD. PMID- 10373753 TI - [Product certification of medical equipment: MedGV (Medical Equipment Act)-MPG (Medical Devices Directive [MDD])]. AB - The changeover from the old MedGV to the new MPG (MDD) may give rise to misinterpretations on the part of the user. The aim of the present article is to introduce the reader to the approval procedures involved. PMID- 10373754 TI - [Vascular endothelial factor (VEGF): therapeutic angiogenesis and vasculogenesis in the treatment of cardiovascular disease]. AB - The formation of new blood vessel is essential for a variety of physiological processes like embryogenesis and the female reproduction as well as pathological processes like tumor growth, wound healing and neovascularization of ischemic tissue. Vasculogenesis and angiogenesis are the mechanisms responsible for the development of the blood vessels. While angiogenesis refers to the formation of capillaries from pre-existing vessels in the embryo and adult organism, vasculogenesis, the development of new blood vessels from in situ differentiating endothelial cells, has been previously considered restricted to embryogenesis. Recent investigations, however, show the existence of endothelial progenitor cells (EPCs) in the peripheral blood of the adult and their participation in ongoing neovascularization. Molecular and cell-biological experiments suggest that different cytokines and growth factors have a stimulatory effect on these bone-marrow derived EPCs. Results with GM-CSF (granulocyte macrophage-colony stimulating factor) and VEGF (vascular endothelial growth factor) open a new insight into the clinical use of cytokines and in particular the use of growth factors in gene therapy. The administration via protein or plasmid-DNA for neovascularization seems to enhance both pathways, angiogenesis and vasculogenesis. PMID- 10373755 TI - [Molecular analysis of the human "growth hormone secretagogue"-receptor]. AB - BACKGROUND: Growth hormone secretagogues (GHS) are highly potent synthetic peptides which release growth hormone (GH) by activation of a growth hormone releasing hormone-independent signal cascade. A specific growth hormone secretagogue receptor (GHS-R) has been isolated, its endogenous ligand is still unknown. It might represent another major endocrine pathway controlling GH secretion. To gain insight into the specific function of the human GHS-R we studied the gene structure. Two variants, type 1a and 1b, have been described, but their specific functions are unknown. METHODS AND RESULTS: A specific probe for the GHS-R was cloned following reverse transcription and PCR amplification of pituitary mRNA. A genomic human placenta library was screened for the GHS-R gene. Positive clones were identified and further characterized by Southern blotting and sequencing. A genomic clone of 18 kb in size was determined to include the coding sequence of both GHS-R variants. Here we show that GHS-R type 1a and type 1b are encoded by a single gene. Sequencing of the immediate 5'-flanking region suggests a number of transcription factor binding sites, but their functional significance remains to be investigated. CONCLUSION: A genomic clone encoding for the two known variants of the human GHS-R was isolated. Further studies will determine physiological relevance and regulation of GHS-R. This will facilitate studies of GHS as diagnostic and therapeutic agents in GH disorders. PMID- 10373757 TI - [Regulation of endothelial NO production by Rho GTPase]. AB - Endothelial-derived nitric oxide (NO) is an important mediator of vascular function. Clinical studies indicate that HMG-CoA reductase inhibitors (statins) improve endothelial function and reduce the incidence of stroke and myocardial infarction. Treatment of human endothelial cells with statins increased the expression of endothelial NO synthase (eNOS) protein and mRNA expression. Statins increased eNOS mRNA half-life but did not change eNOS gene transcription. Inhibition of mevalonate synthesis by statins not only blocks the formation of cholesterol but also of isoprenoids. The upregulation of eNOS expression by statins was independent of cholesterol but mediated via the inhibition of the isoprenoid geranylgeraniol, whereas farnesiol had no effect on eNOS. Immunoblot analyses, (35S)-GTP gamma S-binding assays and transfection studies revealed that statins upregulate eNOS expression by blocking the geranylgeranylation of the GTPase Rho which is necessary for its membrane-associated activity. Studies with mice showed, that statin treatment upregulates eNOS expression and function independent of serum cholesterol levels. Prophylactic treatment with statins augmented cerebral blood flow and reduced cerebral infarcts in normocholesterolemic mice. These effects of statins were completely absent in eNOS-deficient mice indicating that enhanced eNOS activity by statins is the predominant mechanism by which these agents protect against cerebral injury. Our results suggest that statins provide a novel prophylactic treatment strategy for increasing blood flow and reducing brain injury during cerebral ischemia. Upregulation of eNOS by inhibiting Rho may provide a new pharmacologic target for the treatment of arteriosclerosis, pulmonary hypertension, and heart failure. PMID- 10373756 TI - [The 8p11 myeloproliferative syndrome]. AB - CLINICAL MANIFESTATIONS: The 8p11 myeloproliferative syndrome is characterized by a chronic myelogenous leukemia-like myeloid hyperplasia, marked eosinophilia and a strikingly high incidence of non-Hodgkin's lymphoma, mostly of the T lymphoblastic subtype. After a short chronic phase of 6 to 9 months it rapidly transforms into an acute myelogenous leukemia. The median survival time is less than 12 months. CYTOGENETICS: The leukemic cells of peripheral blood/bone marrow and the lymphoma cells have the same acquired, clonal abnormality of chromosome band 8p11 with the translocations t(8;13)(p11;q12), t(8;9)(p11;q34), and t(6;8)(q27;p11). MOLECULAR GENETICS: The molecular cloning of these translocations has shown the fusion of three unrelated genes (ZNF198 at 13p12, FAN at 9q34 and FOP at 6q27) to the fibroblast growth factor receptor-1 (FGFR1) gene at 8p11. The complete coding sequence of the tyrosine kinase domain of FGFR1 is retained in all three fusion genes and presumably activated by sequences of the different fusion partners by dimerization. CONCLUSION: Activation of tyrosinc kinase signal transduction pathways are of increasing interest in the pathogenesis of chronic myeloproliferative disorders and myelodysplastic syndromes. The development of tyrosine kinase inhibitors could represent a promising therapeutic tool. PMID- 10373758 TI - [Aptamers: a novel approach to intervention studies and development of novel therapeutic approaches]. AB - A rapidly growing number of factors is identified that might contribute to disease. To characterize the pathogenetic relevance of a particular factor, specific intervention studies in vivo appear necessary. The present discussion deals with a new class of inhibitors, i.e. aptamers. Aptamers (derived from the latin word "aptus" = fitting) are short DNA or RNA oligomers which can bind to a given ligand with high affinity and specificity due to their particular three dimensional structure and which may thereby, for example, antagonize the biological function of the ligand. Aptamers have been generated against a large variety of molecules ranging from amino acids to complex proteins and even disaccharides. Using platelet-derived growth factor (PDGF) and an experimental mesangioproliferative glomerulonephritis as a model, we describe the in vivo effects of an antagonistic aptamer against PDGF-B. Such studies will greatly aid the identification of the biological role of particular mediators and ultimately the design of novel therapeutic strategies. PMID- 10373759 TI - [Brain metastases of lung cancer: diagnostic accuracy of positron emission tomography with fluorodeoxyglucose (FDG-PET)]. AB - The value of FDG-PET in oncology is currently investigated in clinical studies. There is only limited information on the usefulness of FDG-PET in the evaluation of distant metastases of lung cancer. The purpose of the present prospective investigation was to determine the diagnostic accuracy of FDG-PET in the detection of brain metastases of lung cancer. After intravenous injection of 220 +/- 50 MBq F-18-deoxyglucose PET acquisition was carried out using an ECAT ART scanner (CTI Siemens). Images were reconstructed using a filtered backprojection with a Hanning filter. PET data were analyzed by visual interpretation of coronal, sagittal and transversal slices. PET scans were interpreted by two experienced nuclear medicine physicians without prior knowledge of the results of other imaging studies or clinical data. Between March 1997 and July 1998 whole body PET was performed in 417 patients with suspected lung cancer. 402 patients were used for statistical analysis. Based on conventional brain imaging with CT (occasionally MRI), brain metastases were suspected in 17 patients (prevalence 4.2%). For FDG-PET alone, sensitivity was 82% (14/17) and specificity 38% (14/37). Therefore, diagnostic accuracy of FDG-PET in detection of brain metastases was 93.5%. The low specificity of FDG-PET can be explained by reduced tracer uptake mainly due to brain infarction or vascular encephalopathy in this group of elderly patients. Our results indicate that due to its low specificity FDG-PET is not useful for the evaluation of brain metastases in the primary staging of patients with lung cancer. PMID- 10373760 TI - [Analysis of gene expression patterns in rheumatoid synovial fibroblasts using RAP-PCR for differential display]. AB - OBJECTIVE: Destruction of articular cartilage and bone by invading synovial fibroblasts is a typical histopathologic feature in rheumatoid arthritis (RA). However, little is known about specific up- or downregulation of genes leading to this aggressive phenotype. Thus, our aim was to identify genes, which are differentially expressed in RA synovial fibroblasts as compared to synovial fibroblasts derived from patients with osteoarthritis (OA) using RAP-PCR for differential display. METHODS: After extraction of total RNA, the first step of RAP-PCR was performed using various different arbitrary 10-12-base primers for first-strand cDNA synthesis. Second-strand synthesis was achieved by cycling at low stringency conditions for 35 cycles using different arbitrary 10-base primers, followed by electrophoretic separation and sequence analysis of the amplified fingerprint products. RESULTS: On average, approximately 70 different RNAs were obtained per primer, of which most were expressed both by RA and OA synovial fibroblasts. Using 26 different primer combinations, in total 12 cDNAs were differentially expressed between RA and OA synovial fibroblasts. In the RA group strong amplification of distinct PCR products suitable for sequencing could be observed. Sequence analysis identified these PCR products as highly homologous to various genes involved in regulation of cell cycle and metabolism. CONCLUSION: The data indicate that RAP-PCR is a suitable method to identify differentially expressed genes in rheumatoid synovial fibroblasts potentially involved in the specific pathophysiology of RA. PMID- 10373761 TI - [ZAP genes: characterizing the protein structure of a new family of proliferation associated genes in the exocrine pancreas]. AB - While interested in proliferation-dependent gene regulation in a pancreatic carcinoma cell line, we cloned a set of proteins (ZAP) characterized by a conserved region consisting of consecutive zinc finger, ankyrin repeat and PH domains. Functional aspects of these domains were obtained by comparison with proteins involved in several signal transduction pathways and cell cycle regulation. The members of the ZAP protein family are individually characterized by different types of supplementary protein domains, their chromosomal localization and their tissue specific gene transcription. All results indicate a wide spectrum of protein-protein interactions. Up to now specific binding partners have not been identified. In summary, the multiplicity of conserved regions and transcriptional data indicate a scaffold function for ZAP proteins in the complex network of proliferation associated intracellular signal transduction pathways. PMID- 10373762 TI - [Training psychotherapy]. PMID- 10373763 TI - [Factitious disorders and Munchausens's syndrome. The state of research]. AB - .5-2% of patients of general medicine suffer from factitious disorders. These disorders require utmost skill in recognition, diagnosis and treatment; inadequate treatment may result in severe complications. Literature has grown tremendously during recent years, but many physicians are still not adequately aware of the specific problems. The current state of research concerning phenomenology, pathogenesis, psychopathology, psychodynamics and therapy is presented and discussed. PMID- 10373764 TI - [Psychoanalysis, psychoanalytic psychotherapy and supportive psychotherapy: current controversies]. AB - The author in this paper discusses conceptual, clinical and educational issues of the contemporary controversies regarding the relationship between psychoanalysis, psychoanalytic psychotherapy and psychodynamically oriented supportive psychotherapy. He argues in favor of a strict definition of techniques as the best way leading to a clear-cut differentiation of the three modalities of treatment. Within this frame he discusses interpretation, transference analysis and technical neutrality as the most important aspects and their respective use in the various psychodynamic therapies. Finally he emphasizes that teaching in psychoanalytic and supportive psychotherapies should be part of the psychoanalytic training program of psychoanalytic institutes. PMID- 10373765 TI - [A multicenter study of expenditure and success in psychodynamic therapy of eating disorders. Study design and initial results]. AB - Evaluation of psychodynamically oriented treatment of anorectic and bulimic patients, taking into account both quality and cost of therapist ("How much therapy is applied to which patients for a successful treatment?"). Criteria for indication of the kind of therapy and of amount of therapy ("dosage") are provided. METHOD: Prospective naturalistic longitudinal study (comparable to phase IV of effectiveness studies on pharmaceuticals) including 2.5 year follow up. Consecutive sample (Anorexia nervosa and bulimia nervosa, DSM-III-R). Multidimensional outcome measures including self-rating and expert-ratings. Operationalisation of outcome is layered. Differentiated measurement of amount of therapy and treatment elements. Survival analysis, logistic regression. RESULTS: 1171 completely documented treatment episodes of 43 institutions (30.3% Anorexia nervosa, 55.3% bulimia nervosa, 14.4% double diagnoses). Patients are chronically ill (M = 7.6 years; SD = 6.3 years) and at the beginning of (mostly inpatient) treatment of the index episode 25.5 years of age (SD = 6.0 years). Therapies last appr. 11 weeks (median) and encompass a broad range of therapy measures. DISCUSSION: Study conduction as well as data collection and analysis of this worldwide largest study on treatment of eating disorders is discussed by presenting preliminary results. PMID- 10373766 TI - [Influencing factors of different durations of behavioral therapy in in-patients with anxiety disorders]. AB - In this study, the influencing factors of different durations of in-patient therapy were examined. We investigated 1173 patients with anxiety disorders who were treated in the psychosomatic clinic of Bad Pyrmont with behavioural therapy. Patients with anxiety disorders have an average treatment duration of 52.4 days with a minimum of less than one week and a maximum of more than five months. The duration of treatment depends on the one hand on sociodemographic variables such as age, marital status, partnership, residential status, and professional activity, but not on gender or education. The duration of treatment also depends on factors of severity of the disorder, such as the number of in-patient or psychotherapeutic treatments, duration of disorder and disability, and the number of psychiatric diagnoses, but not on the number of somatic diagnoses. There is a positive correlation between duration of treatment and effect of therapy as well as recovery of fitness for work. PMID- 10373767 TI - [1-Year follow-up of inpatient treatment in a psychosomatic rehabilitation clinic with either a psychoanalytic or behavior therapy oriented treatment]. AB - In a naturalistic setting, we studied the long-term effects of an inpatient treatment program at a psychosomatic rehabilitation clinic, where patients were assigned to a department with either a psychoanalytic or behavior therapeutic orientated treatment after one week of comprehensive diagnosis. The study is based on a self-developed questionnaire which gathers retrospective information in a broad spectrum of problem areas at the point of admittance and release (t0 and t1) as well as current assessments one year following treatment (t2). Additionally, a symptom checklist (Giessener Beschwerdebogen) and a questionnaire assessing depression (Allgemeine Depressionsskala) were prospectively employed at t0 and t2. We asked a consecutive sample of 376 patients to participate, of which 56% answered at t2. Despite limitations of the internal and external validity of the study, we were able to show that substantial improvements are maintained following treatment for both therapies at one-year follow-up, whereas only small difference are found between the outcomes of the two different schools implemented in the clinic. PMID- 10373768 TI - [Psychoanalysis during the Nazi era. Contemporary consequences of a historical controversy: the Wilhelm Reich "case"]. AB - The paper sheds light on the extent of collaboration between the pre-World War II German Psychoanalytic Society (DPG) and the Nazi regime. This is shown by the story of the expulsion of Wilhelm Reich from membership in the DPG, at Freud's own bid. A leading German psychoanalyst, Carl Muller-Braunschweig, published the paper "Psychoanalysis and Weltanschauung" in the fanatically "national" (so called "volkisch") Nazi propaganda organ Reichswart in 1993 following consultations with officials of the International Psychoanalytic Association (IPA) who endorsed these policies. This paper by Muller-Braunschweig was used both to prevent the possible outlawing of psychoanalysis by the Nazis and to deny official DPG support to Wilhelm Reich and the group of leftist-oriented IPA analysts who joined forces with him in opposing Nazi ideology. The paper concludes with examples from post-1945 historiography showing how the exclusion of Reich and the related DPG/IPA compromise and "appeasement" policy were either ignored or disclaimed. PMID- 10373769 TI - [Repercussions in clinical practice of the results of the HOT (Hypertension Optimal Treatment) study]. AB - The HOT study (Hypertension Optimal Treatment) was designed to answer three questions: What is the optimum target diastolic blood pressure (DBP) level that treatment should reach for the greatest reduction in cardiovascular mortality risk; whether there are additional benefits with a progressive reduction of diastolic blood pressure from 90 to below 80 mmHg; and whether low-dose acetylsalicylic acid is of any additional benefit in the primary prevention of myocardial infarction in treated hypertensive patients? To resolve this posing, more than 19,000 essential hypertensives were recruited in 26 countries worldwide by 1,904 investigators with a follow-up period of almost 4 years. Patients were randomly assigned to three different diastolic blood pressure targets: < or = 90 mmHg, < or = 85 mmHg or < or = 80 mmHg respectively. Additionally, in each group, patients were randomly assigned to acetylsalicylic acid or placebo. The follow-up period was about 4 years. Although the HOT trial failed to demonstrate the primary objective for which it was designed, of defining optimum target diastolic blood pressure to be achieved by antihypertensive treatment, this HOT study has helped to clarify very important issues related to blood pressure lowering such as the nonexistence of the J-shaped curve concept in hypertension, the possibility of achieving substantial improvement in blood pressure control by the use of antihypertensive combination therapy, the additional benefit of low-dose aspirin in primary prevention of myocardial infarction among treated hypertensives, and the demonstration of the necessity of reducing diastolic blood pressure below 80 mmHg in diabetic hypertensive patients. For all these reasons, the HOT study is destined to become among the most relevant hypertension intervention studies of this century. PMID- 10373770 TI - [New guidelines from the World Health Organization and the International Society of Hypertension for the management of hypertension: toward common guidelines]. PMID- 10373771 TI - [Intracoronary ultrasound: A necessary tool for stent implantation? Arguments in favor]. AB - Intracoronary ultrasound (ICUS), as opposed to angiography, provides high resolution, tomographic images of the coronary vessel and lumen. Because of its superior diagnostic sensitivity ICUS is indicated in the evaluation of suboptimal results and complications following stent implantation. Only a few years ago the use of stents was limited by a high incidence of subacute thrombosis. ICUS demonstrated that the deployment technique used at that time was inadequate and that stent expansion could be improved by the routine use of high pressure inflation, leading to a simplification in the anticoagulation regimen and a decrease in the subacute thrombosis rate in elective procedures to < or = 1%. However, the routine use of high balloon pressures does not assure an adequate expansion of the stent. Only about one third of the stents deployed under angiographic guidance are optimally expanded, with intra-stent luminal dimensions similar to the adjacent, reference, luminal sizes. Significantly, these underdeployed stents can be recognized by ICUS and a large proportion adequately expanded. It should be emphasized that the best predictors of stent restenosis are two ICUS parameters, the postprocedural luminal dimensions and the % cross sectional narrowing, and not the angiographic parameters. Likewise, two of the lowest restenosis rates ever reported (12.8% and 7.3%) have occurred in two studies (WEST-2 and MUSIC) in which stent deployment was guided by ICUS. Two trials (AVID and OPTICUS) have been specifically designed to test the hypothesis that routine use of ICUS to guide stent implantation could diminish the restenosis rate, but their final results are not yet available. The CRUISE study was designed to evaluate the impact of routine ICUS not on angiographic restenosis but on the clinical need of revascularization. In this trial, the larger luminal dimensions of the stents implanted under ICUS guidance translated into a 40% reduction in the 6 month revascularization rate (14.8% vs. 8.9%, p < 0.05). Although the final answer is still pending, the available information suggests that the routine use of ICUS might translate into a direct clinical benefit, something remarkable for a diagnostic tool. In any case, the most effective way of using ICUS would probably be identifying those lesions that most benefit from the technique and avoiding its use in lesions with, a priori, excellent results. PMID- 10373772 TI - [Intracoronary ultrasound: A necessary tool for stent implantation? Arguments in against]. AB - Intracoronary ultrasound has shown the little reliability of angiography to predict the interaction of stents with the arterial wall. In spite of implanting the stents with high pressures and a good angiographic result, a great proportion are still incompletely expanded. The use of intracoronary ultrasound as a guide for stent implant allows us to optimize the degree of expansion and apposition of the stent to the arterial wall, achieving greater intraluminal dimensions than with angiography. Nevertheless, this strategy is not necessarily translated in a clear clinical benefit. The rate of acute and subacute complications of stents implanted under angiographic control with high pressures and treatment with ticlopidine and aspirin is less than 1% and identical to the studies that use intracoronary ultrasound to optimize stent deployment. At the present time, it has not been documented either that the optimization of stent deployment with intracoronary ultrasound significantly reduces the rate of restenosis, the incidence of target vessel revascularization or the rate of major adverse cardiac events in mid-term follow-up. In addition, the use of intracoronary ultrasound to optimize stent deployment adds a small risk to the procedure, extends the time of occupation of the cardiac catheterization laboratory and prohibitively increases the costs of coronary stenting them being already high ones. Thus, the universal use of coronary intracoronary ultrasound to optimize stent deployment seems not to be, at present, a useful strategy. PMID- 10373773 TI - [Catheter ablation of accessory pathways in infants and children weighing less than 10 kg]. AB - INTRODUCTION: Low weight is considered an independent risk factor for the appearance of complications in radiofrequency catheter ablation of accessory pathways in children. OBJECTIVES: The purpose of this study was to evaluate the results and long term follow-up of radiofrequency catheter ablation in eight infants and small children of accessory pathways of less than 10 kg in weight. METHODS AND RESULTS: There were 3 boys and 5 girls with a mean age of 6.3 +/- 5 months (range, 2.5 to 17) and an average weight of 6.2 +/- 1.9 kg (range, 3.5 to 9). The eight patients underwent a single successful ablation procedure. Five left free wall pathways were ablated by transseptal approach, two right posteroseptal pathways were ablated from the inferior vena cava and a left posteroseptal was approached from the inferior vena cava into the coronary sinus. A deflectable 5F bipolar electrode catheter with a 3 mm tip was used in the first five patients and a deflectable 5F tetrapolar catheter with a 4 mm tip and temperature monitoring using closed loop control in the 3 remaining patients. An abrupt increment in impedance due to the development of a coagulum was observed in 2 procedures. One patient developed an acute ischemic complication during ablation of a left lateral accessory pathway by transseptal approach. This patient had mild pericardial effusion after the procedure. Moderate pericardial effusion was also noted in another patient after radiofrequency ablation that resolved itself spontaneously. In the remainder of the procedures there were not complications. After a mean follow-up of 32.3 +/- 22.1 months (median 42) all patients are asymptomatic without antiarrhythmic treatment. CONCLUSIONS: a) radiofrequency catheter ablation can be performed successfully in infants and small children weighing less than 10 kg, and b) echocardiography must be performed inmediately after the procedure in infants to investigate pericardial effusion. PMID- 10373774 TI - [The etiology and associated risk factors in a sample of 300 patients with atrial fibrillation]. AB - PURPOSE: To analyze the etiology and the prevalence of risk factors in patients with atrial fibrillation. PATIENTS AND METHODS: Applying an unpaired case controlled study, we examined 300 consecutive patients (143 men) with atrial fibrillation and a mean age of 66 +/- 8 years. This group is compared with a control group of 700 patients (mean age 64 +/- 12 years). RESULTS: In the group with atrial fibrillation the etiology in 32% was arterial hypertension, in 20% coronary heart disease, in 13% valvular heart disease, in 11% heart failure, in 4% hyperthyroidism and in 20% idiopathic. 50% presented hypertension, 29% tobaccoism, 26% left ventricular hypertrophy, 20% consumption of alcohol, 19% hypercholesterolemia and 16% diabetes. Compared with the control group, patients with atrial fibrillation had coronary heart disease (p < 0.05), VHD (p < 0.01), myocardiopathy (p < 0.05), HT (p < 0.001), left ventricular hypertrophy (p < 0.001), diabetes (p < 0.01) and alcohol consumption (p < 0.01) more frequently. In the multivariant analysis heart failure (odds ratio 2.1 [1.2-3.3]), the valvular heart disease (odds ratio 2.2 [1.4-3.5]), the coronary heart disease (odds ratio 1.8 [1.2-2.6]), the arterial hypertension (odds ratio 1.7 [1.2-2.3]), the left ventricular hypertrophy (odds ratio 2.6 [1.7-3.8]), the diabetes (odds ratio 1.9 [1.2-2.9]) and alcoholic habits (odds ratio 2 [1.3-3.9]) were independent risk factors for atrial fibrillation in our population. CONCLUSIONS: Atrial fibrillation in our study, is more frequent in patients with arterial hypertension, coronary heart disease or valvular heart disease. There are other risk factors such as arterial hypertension, diabetes and consumption of alcohol too, the modification of which could diminish the risk of the appearance of atrial fibrillation. PMID- 10373775 TI - [Feasibility and safety of intracoronary ultrasound. Experience of a single center]. AB - BACKGROUND AND OBJECTIVES: Intracoronary ultrasound provides a number of advantages in the quantification and characterization of coronary stenoses with regard to contrast angiography. However, previous studies have reported a 3.5 to 11% complication rate, and a 10-30% failure rate in performing this technique. The purpose of the study is to analyze the feasibility of performing intracoronary ultrasound and the incidence of complications associated with the use of contemporary, state of the art equipment. MATERIAL AND METHODS: The feasibility of performing intracoronary ultrasound, analyzed as the percentage of successes and failures in performing the examination was reviewed, as well as the complication rate associated with the technique in all the procedures carried out between July 1, 1994 and February 29, 1996 in which intravascular ultrasound was attempted. Complications were categorized as related, non-related and uncertainly related to the ultrasound study. RESULTS: 239 vessels were studied with intravascular ultrasound in 209 procedures (74% interventional) performed on 139 patients. Ultrasound examination was feasible in all the diagnostic studies and in 96% of the interventional procedures. The major and minor procedural complication rate was 2.4 and 10.5% respectively. No major complication was related to the ultrasound examination. Three patients experienced minor complications (1.4%) related to the ultrasound study. All three complications occurred in baseline studies during interventional procedures. CONCLUSIONS: Intracoronary ultrasound is feasible and safe in the vast majority of the procedures. Improvements in smaller catheter size and design and larger operator expertise have significantly reduced the complication rate, particularly the most frequent coronary spasm so far. Complications are associated with baseline studies during interventional procedures and with less operator expertise. PMID- 10373776 TI - [Expression of inducible nitric oxide synthase in the carotid of rats after endothelial skinning: the effects of platelets and treatment with abciximab]. AB - BACKGROUND: Functional evidence suggests that endothelial denudation stimulates inducible nitric oxide synthase (iNOS) activity in the vascular wall. In vitro studies done in our laboratory have shown that iNOS expression in smooth muscle cells is reduced by endothelial cells. The object of this study was to analyze the iNOS protein expression in the arterial wall after in vivo deendothelialization, and the role of platelet activation abciximab in the expression of this protein. MATERIALS AND METHODS: Endothelial denudation was performed in the left carotid artery of Wistar rats. The right carotid artery was used as control. RESULTS: iNOS protein was only weakly expressed at 6, 24 and 48 hours after endothelial denudation. Since platelet adhesion and aggregation occur early after endothelial damage, we have analyzed the role of activated platelets in iNOS protein expression during the first two days after angioplasty. Early after in vivo endothelial injury, thrombocytopenic rats showed a marked iNOS protein expression. Similar results were obtained by blocking the platelet glycoprotein IIb/IIIa in rats treated with abciximab (Reopro). CONCLUSIONS: iNOS protein is weakly expressed in the arterial wall after endothelial denudation. Platelets play a crucial role preventing iNOS protein expression early after endothelial damage through a mechanism that depends on GP IIb/IIIa, an effect that can be avoided with glycoprotein IIb/IIIa, blockers, such as abciximab. PMID- 10373777 TI - [Ischemic preconditioning. Is it always a beneficial phenomenon?]. AB - INTRODUCTION AND OBJECTIVES: Hearts exposed to reversible ischemia stand a subsequent prolonged episode of coronary artery occlusion (ischemic preconditioning) better. The reduction of infarct size by means of preconditioning has been amply demonstrated, but the relationship between preconditioning and contractile function remains less well defined. In this study we assess the effect of a later ischemia on the regional contractility in a stunned-preconditioned myocardium. METHODS: We analyze the shortening fraction in the ischemic (dependent on the left anterior descending coronary artery), periischemic and control zone (dependent on the left circumflex coronary artery), using chronic implants of ultrasonic crystals in 17 adult mongrel dogs. In the control series, we quantified the effects of partial (30-60% reduction of coronary flow from the basal) and transitory (15 minutes) ischemic episode in the regional myocardial function in a "virgin" myocardium. In two other series, the myocardium was previously stunned-preconditioned through brief and repeated ischemias. Afterwards, at 5th day (series B) and at 15th day (series C), the dogs were subjected to ischemic episode similar to control ones. RESULTS: After comparing the results with the control series, we observed that the shortening fraction of the ischemic zone was decreased by 107% (p < 0.01) during partial ischemic episode when it was induced on the 5th day of the stunning preconditioning (series B). CONCLUSIONS: In dogs, the brief and repeated episodes of ischemia could condition the contractile function so that a later partial and transitory reduction of coronary flow could induce a severe affectation of contractility expressed as a diskinetic area. PMID- 10373778 TI - [Improvement of myocardial perfusion after transmyocardial laser revascularization]. PMID- 10373779 TI - [Progressive subaortic stenosis caused by accessory mitral valve tissue]. AB - Accessory mitral valve tissue is a rare cause of subaortic obstruction. The reported case correspond to a two days old patient diagnosed by 2D Echocardiography. Serial doppler flow analysis showed progressive left ventricular outflow obstruction. A cardiac catheterization at the age of 9 months confirmed the obstruction without defining the cause. Five months later, transesophageal echocardiography clearly defined the intraventricular connections of the accessory mitral tissue. At the age of 18 months the patient suffered from cardiac failure and underwent surgery for removing the accessory tissue, assisted by intraoperative transesophageal echocardiography pre and post by-pass. Follow up echocardiography at 25 months showed no left ventricle outflow obstruction and the patient is now asymptomatic. PMID- 10373780 TI - [Myocardial bridging as a cause of acute ischemia. Description of a case and review of the literature]. AB - Myocardial bridges consist of muscle fiber bundles lining an epicardial coronary artery for a variable distance. They are a relatively common finding, with incidence changing on the basis of the study method used (angiographic/necropsy). Although myocardial bridges are usually associated with a benign prognosis, being in many cases asymptomatic and only found by chance, their presence has also been considered a cause of angina, malignant arrhythmia, myocardial infarction and sudden death. They are diagnosed in vivo by angiography when a systolic compression of a coronary artery which disappears during diastole is evidenced. We report the case of a patient with electrocardiographic signs of severe ischemia in the territory of the anterior descending artery, which was initially assessed as myocardial infarction and treated as such. Eventually, the ECG returned to normal, and no new Q waves of necrosis occurred. An angiohemodynamic study confirmed the existence of an isolated muscular bridge over the middle third of the anterior descending artery, with no other associated coronary lesions. PMID- 10373781 TI - [Coronary surgery with minimum access and cardiopulmonary bypass]. AB - From October 1997 to March 1998 we operated on seven patients with minimal incision, cardiopulmonary by-pass with femoral cannulation and antegrade blood cardioplegic arrest using the "endoclamp" (Heartport Inc.). The seven patients with isolated severe lesions of the left anterior descending underwent a left internal thoracic artery graft under direct vision. Three had saphenous vein coronary bypass grafts performed to the diagonal (2) and obtuse marginal branches of the left coronary artery. The median cardiopulmonary bypass duration was 75 minutes (30-230) and the aortic occlusion time was 33 minutes (10-117). No major complications occurred and only two minor ones were noted. The median intensive care unit stay was 2 days (1 to 4) and the total hospital stay was 6.5 days (3 to 13). All the patients are in NYHA FC I, without treatment and a follow up of 3 to 6 months after the surgery. With this method of myocardial revascularization with minimal incision and cardiopulmonary bypass the sternotomy-related complications can be avoided, the intensive care unit and hospital stay can be reduced with better convalescence for the selected patients. We believe that this technique is a valid option for an increasing number of patients. PMID- 10373782 TI - [Intravascular hemolysis following percutaneous occlusion of the ductus arteriosus]. AB - Transcatheter occlusion of patent ductus arteriosus has become a safe and successful technique, but it's not free of complications. We present the case of a two-year-old boy who underwent routine transcatheter closure of his patent ductus arteriosus, using a "coil" device. Twenty hours later he developed severe persistent hemolysis in association with residual ductal flow. Patient's clinical situation became stable when the device was removed. Pulmonary embolization of the device and hemolysis are the main complications of percutaneous closure of the patent ductus arteriosus. Hemolysis occurs rarely (0.5%) and is always associated with the presence of residual ductal flow. Several approaches to this problem have been described. Mild cases may require no intervention; however, when severe hemolysis is present, removal of the device may be needed, proceeding with surgical repair of the patent ductus arteriosus. PMID- 10373783 TI - [Aortic dilation and dissection in a patient with Turner syndrome]. AB - The ascending aortic dilatation and its dissection is a not very frequent finding in patients with Turner syndrome. The high incidency of structural anomalies in the aortic wall and the severity of its complications, makes it necessary to watch these patients very closely. We present an asymptomatic patient, affected with Turner syndrome, ascending aortic dilatation and aortic wall dissection. PMID- 10373784 TI - [Benign atheroembolic syndrome secondary to systemic fibrinolysis with streptokinase]. AB - We present the case of an 60-year-old male patient that after a acute myocardial infarction inferior diagnostic was subjected to a fibrinolysis with streptokinase. The evolution from the cardiologic point of view was favourable, but at fourth day postacute myocardial infartion he starts with peripherics vasculars symptoms, pain and livedo reticularis in lower part of the body. A symptomatic treatment was made requiring amputation of his fifth right toe. The evolution was towards healing without any organic afectation at another level. PMID- 10373785 TI - [The Ross technique in Spain]. PMID- 10373786 TI - [Guidelines for the diagnosis and management of heart failure and cardiogenic shock. Informe del Grupo de Trabajo de Insuficiencia Carddiacade la Sociedad Espannola de Cardiologia]. AB - Guidelines for the Diagnosis and Management of Heart Failure and Cardiogenic Shock have been developed by the Working Group on Heart Failure of the Spanish Society of Cardiology, in collaboration with other Scientific Sections and members of the society. The aim of this report is to promote a more consistent and effective clinical practice according to the principles of evidence based medicine or the recommendations widely accepted by the scientific community. At the same time the aim is to give guidance for epidemiological surveys, heart failure registers clinical assays and clinical quality assessment, and to contribute to cost containment. These twelve guidelines have been designed for doctors in general practice as well as specialists. Criteria for diagnosis and classification of heart failure (systolic and diastolic heart failure, left or right, acute or chronic) are defined. The more appropriate use of clinical or high technology laboratory studies are recommended as well as the most efficient strategies nowadays for the management of chronic stable, unstable or refractory heart failure, or acute heart failure and cardiogenic shock. PMID- 10373787 TI - [How important are attitudes towards washroom hygiene and spitting in schools? A telephone survey of school attendants in the Baselstadt canton, April 1998]. AB - The aim of the study was to evaluate the risk spitting poses for students at Basle schools and to document the state of their toilet facilities. A telephone survey with 43 caretakers of the 56 Basle schools revealed, that spitting was considered a problem at 75% of the schools and that at every 4th school it had recently gotten worse. Apart from spitting on the floor (93%), spitting at walls and windows (32%), doors (20%), mirrors (16%), and occasionally at other persons (14%) occurred. Just about every school was equipped with the advised number of toilet units and urinals. On the other hand, 29% of handdrying mechanisms and 36% of soap-type offered as well as 39% of cleaning frequencies of facilities did not correspond to advised norms. 93% of toilet flushing mechanisms, 38% of the urinals and 96% of the washbasins had to be operated manually. This survey shows that spitting was common at practically every Basle school. Spitting at each other which is risky for infection occurred only sporadically. The risk of infection by spitting probably plays a minor role compared to other routes of infection. Despite this we advise a renewed effort in informing parents and students. Toilet hygiene at Basle's schools is still deficient. The facilities should be improved by midterm investments. PMID- 10373788 TI - [Hospital physicians in the light of letters of complaint]. AB - In a competitive health-care system the quality of services and, closely related, customer satisfaction are significant success factors. Some patients or their relatives who are dissatisfied with the provided service express this in letters of complaint. Using the critical incident technique based on a sample of 54 letters of complaint regarding doctors' performance sent to the administration of the Inselspital of Berne we examined information such feedback contains. A total of 133 critical occurrences have been described. Seventy percent of the occurrences were related to medical treatment and 6% to the medical education and research process. The remaining 26% of the complaints concerned accommodation, care, administration or other areas. In the sphere of medical service procedures the dissatisfaction of patients centered on information provided to patients and their families (38%), followed by criticism concerning clinic organization (23%), interaction with the doctor (14%), his professional competence (13%) as well as the management of clinical data (12%). On average, 3 out of 10 letters allude to possible criminal or liability proceedings or seemed to have a potential for escalation in other areas. The situations as presented in these complaints is to be considered as objective and relevant. To continue the process of optimization and quality improvement it is therefore recommendable to take these signals seriously. PMID- 10373789 TI - [The end of handwashing: to the next millennium with hygienic disinfection of hands]. PMID- 10373790 TI - ["I am tired"]. PMID- 10373791 TI - [Powerlessness and apathy in digitalis intoxication]. AB - A 85 year old female was hospitalized because of a bronchopulmonary infection. During the hospitalization she developed a progressive stupor. There was no sign of an intracerebral pathology, an electrolyte disorder, a new infection or a psychiatric diagnosis. Evaluation of the past history made a recently started digitalis medication responsible for the stupor. After discontinuation of digitalis the patient regained complete consciousness within two weeks. In the Holter-EKG we found once an asystole of four seconds duration without any symptoms. PMID- 10373792 TI - [Idiopathic acute panniculitis]. PMID- 10373793 TI - [Unusual cause of hyponatremia]. PMID- 10373794 TI - Veterans Affairs NPs flex muscle. PMID- 10373796 TI - Lobbyists vital to NP success. PMID- 10373795 TI - Writing prescriptions for family. PMID- 10373797 TI - Primary care management of multiple sclerosis. AB - Multiple sclerosis is a chronic disease of the central nervous system that affects young adults most often. The course of MS is unpredictable and variable on a day-to-day basis. As chronic problems accumulate, the disease may become more steadily progressive, with fewer or no acute relapses. A diagnosis of multiple sclerosis is a clinical one based on a thorough history and neurologic examination findings and supported by magnetic resonance imaging of the brain and evoked potential studies. Management strategies fall into three general categories: treatment of relapses caused by underlying disease; prevention of progression or reduction of the frequency of relapses; and control of specific symptoms. PMID- 10373798 TI - Ovarian decline. In the later reproductive years. AB - While most healthy older women who become pregnant have uneventful pregnancies and healthy babies, thousands more are unable to achieve pregnancy because of declining ovarian function. A decline in ovarian function is normal with increasing age. A woman's number of eggs steadily decreases from a peak at mid gestation of 7 million to approximately 400,000 at puberty. Certain tests can reflect gradations in ovarian reserve status and predict a woman's potential fertility. The most important lab test is the day 3 follicle-stimulating hormone level. A constant source of frustration in fertility centers is that patients are referred to the specialty clinic too late, when diminished ovarian reserve is so marked that treatment success is severely limited. Earlier assessment of ovarian function would alter this unfortunate trend. PMID- 10373799 TI - Obesity in children. AB - The most common definition of obesity in childhood is weight in excess of 20% above ideal weight for age and gender and skinfold thickness in excess of the 85th percentile for age and gender. Any treatment plan for overweight children and adolescents should include three major components: diet, exercise and family based behavior management. Children and adolescents should not be placed on restrictive diets because adequate calories are needed for proper growth and development. Aside from calorie excess and inactivity, genetics and environment predispose a child to be obese. Therefore, any program or treatment plan must include the caretakers, who in most cases are also overweight or obese. PMID- 10373800 TI - Prenatal screening. An expanding role for nurse practitioners. PMID- 10373801 TI - Beyond the big cities. HIV in rural America a growing threat. PMID- 10373802 TI - Taming 'valley fever'. PMID- 10373803 TI - Homocysteine and the heart. PMID- 10373804 TI - Breaking new ground in Ireland. Interview by Richard T. Curtin. PMID- 10373805 TI - Making an 'impression'. PMID- 10373806 TI - Seeing red. A review of subconjunctival hemorrhage. PMID- 10373807 TI - Learning from the patient's view. PMID- 10373808 TI - Neonatal cerebral infarction: an under recognised/unreconised cause of neonatal seizures? AB - Neonatal cerebral infarction or neonatal stroke is a well recognised cause of neonatal seizures. The awareness of its existence among healthcare workers is low and it is not even mentioned as a cause of neonatal seizures in most books. Many of the affected neonates are well, without any evidence of neurological disturbances between seizures. Awareness of its existence among healthcare workers is critical for the diagnosis of this problem. Early diagnosis can lead to early intervention, which may mean a better prognosis for these children. PMID- 10373810 TI - Obstetric outcomes in an aboriginal community: a comparison with the surrounding rural area. AB - Antenatal, intranatal and postnatal features of all Aboriginal women who lived at Cherbourg Aboriginal Community and delivered during 1990, 1991 and 1992 were compared with all non-Aboriginal women in the same rural area who delivered at Kingaroy Base Hospital during 1991. Almost all the Aboriginal women also delivered at Kingaroy. The data for 146 Aboriginal and 139 non-Aboriginal women were taken from the hospital records. The Aboriginal women were generally younger at delivery (Aboriginal 35% younger than 20 years vs non-Aboriginal 12%), made their first antenatal visit later (Aboriginal 49% after 20 weeks vs non Aboriginal 10%) and made fewer antenatal visits (Aboriginal 43% < 4 visits vs non Aboriginal 2% < 4 visits). They were more likely to be anaemic (Aboriginal 65% < 110 g/L vs non-Aboriginal 13% < 110 g/L), have a sexually transmitted disease (STD; Aboriginal 13% vs non-Aboriginal 2%) and drink alcohol (Aboriginal 54% vs non-Aboriginal 32%). After making an allowance for repeat Caesarean sections, there was no significant difference in the proportion of abnormal deliveries, but birthweights of Aboriginal infants were lower. Postnatally, the only significant difference between the two groups was a lower incidence of jaundice in Aboriginal infants. Multifactorial analysis showed that birthweights were significantly decreased by primagravidy, alcohol intake and STD. It is likely that the effects of STD and alcohol on birthweight were due to associated lifestyle factors. When these factors were allowed for, ethnic background had no significant effect on birthweight. PMID- 10373809 TI - Evaluation of a self-paced education package on violence against women for rural community-based health workers. AB - There are no reported education programs specifically focusing on the needs of rural health workers in the area of violence against women. The most commonly reported contact sought by women experiencing injuries and health problems associated with violence and abuse is with health workers. Women report a failure by health workers to make direct enquires, which may be due to their lack of education and confidence in responding to these issues. A convenience sample of 60 community-based rural health workers from a range of occupations and settings within the Wide Bay Health Region, Queensland, participated in the evaluation of a self-paced, distance education package on violence against women. The package contained seven modules. These included written and audio tape material, and activities that together formed a community development approach to addressing the needs in the participants' local community. Participants were given a mentor and teleconference support during the 8 weeks allocated to complete the package. A pre- and post-course evaluation, containing quantitative and qualitative data, was completed. Analysis of the quantitative data identified significant changes in participants' knowledge, and the qualitative data highlighted an increased sense of confidence in assisting women, forming support networks and using resources more effectively. Participants reported the most useful aspects of the package were: (i) modular- and user-friendly format; (ii) flexible, practical, health-focused content; and (iii) real world examples. PMID- 10373811 TI - Hospital admission and mortality differentials of asthma between urban and rural populations in New South Wales. AB - It remains unclear whether there are any differentials in hospital admission and mortality rates of asthma between urban and rural populations. An observational study was conducted, based on patient hospital records, to examine the distribution of asthma admissions and mortality in New South Wales. Data on all reported cases of asthma were obtained from New South Wales hospitals between 1989 and 1994. Information on deaths of asthma was collected between 1983 and 1992. The hospital admission rates of asthma varied from 4.8 per 1000 in 1990 to 5.4 per 1000 in 1992 for rural population, and from 3.0 per 1000 in 1991 to 3.4 per 1000 in 1992 for urban population. The hospital admission rates were 51.2 69.1% higher for rural residents than urban dwellers. The mortality rates of asthma ranged from 4.8 per 100,000 in 1983 to 8.0 per 100,000 in 1985 for rural population, and from 3.8 per 100,000 in 1983 to 6.0 per 100,000 in 1989 for urban population. The mortality rates of asthma were 3.62-42.85% higher for rural residents than urban dwellers. These results indicate that the non-age-adjusted hospital admission and mortality rates of asthma were considerably higher in rural populations than in urban populations in New South Wales. PMID- 10373812 TI - Prevalence and sociodemographic determinants of cardiovascular risk in a rural area. AB - Non-metropolitan areas have a higher mortality from cardiovascular disease than metropolitan areas. The study's aim was to establish the prevalence of cardiovascular disease risk factors in a rural area and identify their sociodemographic determinants. Adults, randomly selected from Ballarat's electoral rolls, were invited to complete a questionnaire and have their height, weight, blood pressure and fasting lipids measured. Three hundred and thirty eight eligible persons participated (67% response). The data were analysed using logistic and multiple regression analyses. Increasing age was associated with hypertension, high plasma cholesterol, overweight/obesity, high plasma triglyceride levels and increasing plasma fibrinogen. Women were less likely to be overweight/obese and have a high plasma triglyceride. Not having completed high school was associated with hypertension, high plasma cholesterol and triglyceride levels and physical inactivity. Smoking was associated with employment and being in a non-professional/managerial occupation. Rural health promotion initiatives should take account of the needs of these population subgroups. PMID- 10373813 TI - Psychosocial profile of pregnant adolescents in a large Australian regional area. AB - A psychosocial profile was developed of 122 pregnant adolescents attending a public hospital antenatal clinic in a large regional Australian area. Participants completed the Rosenberg Self-Esteem Scale and the Support Behaviours Inventory and were interviewed to obtain psychosocial and demographic information. Results revealed that those who were most likely to have either or a combination of poor self-esteem, lack of social support, be unemployed or smoke, were more likely to be living alone or with friends, not have a partner or have a partner for a shorter period, have a partner who was older, have previous children, have not planned their pregnancy and have less education. The degree of social support was significantly associated with self-esteem. Over half of the sample smoked, and over half were unemployed. These results underscore the importance of addressing psychosocial factors in the implementation of care for the pregnant adolescent. PMID- 10373814 TI - A study of postdiagnosis breast cancer concerns for women living in rural and remote Queensland. Part I: Personal concerns. AB - The findings presented in this paper are part of a research project designed to provide a preliminary indication of the support needs of postdiagnosis women with breast cancer in remote and isolated areas in Queensland. This discussion will present data that focuses on the women's expressed personal concerns. For participants in this research a diagnosis of breast cancer involves a confrontation with their own mortality and the possibility of a reduced life span. This is a definite life crisis, creating shock and needing considerable adjustment. Along with these generic issues the participants also articulated significant issues in relation to their experience as women in a rural setting. These concerns centred around worries about how their partner and families cope during their absences for treatment, the additional burden on the family of having to cope with running the property or farm during the participant's absence or illness, added financial strain brought about by the cost of travel for treatment, maintenance of properties during absences, and problems created by time off from properties or self-employment. These findings accord with other reports of health and welfare services for rural Australian and the generic literature on psycho-oncology studies of breast cancer. PMID- 10373815 TI - A study of postdiagnosis breast cancer concerns for women living in rural and remote Queensland. Part II: Support issues. AB - This paper presents the recent findings from a study on the postdiagnosis support needs of women with breast cancer living in rural and remote Queensland. The findings presented in this discussion focus on support needs from the perspective of the women experiencing breast cancer as well as health service providers. The tyranny of distance imposes unique hardships, such as separation from family and friends, during a time of great vulnerability for treatment, the need to travel long distances for support and follow-up services, and extra financial burdens, which can combine to cause strains on the marital relationship and family cohesion. Positive indications are, however, that the rural communities operate on strong, informal networks of support. This network of family, friends and community can, and does, play an active role in the provision of emotional and practical support. PMID- 10373817 TI - Rural health status: what do statistics show that we don't already know? AB - Arguably the policies and programs designed to bring about improvements in the health status of rural and remote residents have been limited by the absence of systematic statistical data about health status and its relationship with place of residence. The Australian Institute of Health and Welfare addresses this problem in the recent report Health in rural and remote Australia. The report compares the health status, health risk factors and preventative measures, and health resources for rural, remote and urban areas. The data highlights that people in rural and remote areas of Australia have poorer health status than their metropolitan counterparts on several counts. While further work is required, the report provides a useful and important basis for identifying and monitoring the pattern of health status and resource availability in rural and remote Australia. PMID- 10373816 TI - Postacute care for older aboriginal people: an exploratory-descriptive study. AB - Many Aboriginal people reside in rural and remote Australia. Aboriginal health workers were the informants in this exploratory-descriptive study, which explored issues pertaining to postacute care for older Aboriginal people. Qualitative analysis of interview data revealed several issues were viewed as being of crucial importance in the provision of effective postacute services to older Aboriginal people. These were: (i) identification of Aboriginality; (ii) perceived racism and stereotypical attitudes among hospital staff and healthcare workers; and (iii) effective discharge planning. Other issues which were believed to impact upon service use were identified as: (i) availability of services; (ii) knowledge of services and level of use; and (iii) the notion of mainstream versus Aboriginal-specific services. Findings are discussed in relation to available literature. Implications for further research are drawn from the findings of this exploratory study. PMID- 10373818 TI - Towards a culture of improving indigenous health in Australia. PMID- 10373819 TI - Role of technology in achieving clinical and cost impact in acute and critical care. PMID- 10373820 TI - Advances in artificial airway management. AB - This article has reviewed some of the current airway management practices as well as technologic advances in adult tracheal tube design in the critically ill patient. Although the primary goals of airway protection and facilitation of positive pressure ventilation remain unchanged, a better understanding of the limitations of tracheal tubes, as well as strategies for optimized airway management, is critical. Such information is needed in order to reduce or avoid potential complications associated with tracheal tubes or similar airway devices. PMID- 10373821 TI - Hemodynamic monitoring. AB - Hemodynamic monitoring is one of the most exciting and potentially useful technologies in critical care. Hemodynamic monitoring, particularly the PAC, is the technology most often associated with the critical care unit. However, it is a difficult technology to master and is associated with clear (although infrequent) serious complications. It can also be associated with increased costs. With appropriate implementation, this technology can improve patient outcome and moderately reduce costs. Appropriate implementation is not easy with this technology. This type of technology should only be employed in hospitals willing to invest the education and quality monitoring to ensure its appropriate application. As a part of this infrastructure, physicians and nurses need frequent communication in terms of what is expected from this technology for each patient. PMID- 10373822 TI - Continuous mixed venous (SvO2) monitoring. Too expensive or indispensible? AB - Based on theoretic principles and clear literature support, SvO2 (mixed venous oxyhemoglobin) monitoring offers an important advantage over traditional hemodynamic parameters. SvO2 allows more precise understanding of the adequacy of cardiac and pulmonary function than traditional parameters. SvO2 values do not replace the need to measure individual parameters of oxygen delivery or consumption but serves as the standard for assessing the impact of each parameter on tissue oxygenation. SvO2 has the ability to reflect a threat to tissue oxygenation that is unmatched by other parameters. Its ability to give a real time indication of tissue oxygenation makes it a preferred parameter for monitoring the adequacy of hemodynamics. Its use as an end point for determining the adequacy of hemodynamics (BP, CO/CI), measurement of Qs/Qt, and prediction of potential hemodynamic instability make this parameter invaluable for the knowledgeable clinician. SvO2 catheters cost more than traditional PA catheters, a factor that has limited their acceptance as the standard PA catheter. The added cost of the SvO2 catheter has not been adequately addressed in the research literature. However, research does support SvO2 allows more rapid termination of drug therapies, may improve movement out of the ICU, and reduces the incidence of mechanical ventilator manipulation. These features increase the cost effectiveness of SvO2 catheters. The cost-effectiveness of the catheter, like any technology, is predicated on the clinician. Clinicians++ must be educated to use SvO2 as a primary end point for treatment decisions regarding hemodynamic therapy and patient stability. If used properly, every PAC should use fiberoptic SvO2 capabilities. PMID- 10373823 TI - Capnography. A key underutilized technology. AB - Based on the multiple applications and the potential cost savings, every ICU should have enough capnography for all intubations and probably for all mechanically ventilated patients. Of the multiple clinical applications of capnography, most attention should be focused on its use with intubation and resuscitation. Other applications, such as blood gas and ventilation-perfusion scan reduction, should be instituted after the primary areas have been implemented. While capnography modules may appear to be expensive at first glance, an analysis of their clinical application reveals they can save the hospital hundreds of thousands of dollars beyond the purchase price. PMID- 10373824 TI - Advances in continuous, noninvasive hemodynamic surveillance. Impedance cardiography. AB - In the current climate of shrinking health care reimbursement and increasing importance of patient centered care, impedance cardiography is one method of enhancing quality of patient care and appropriate use of resources. Hemodynamic and thoracic fluid status data may be obtained quickly, accurately, and without risk, providing a global clinical perspective. Patients benefit from the ability to immediately obtain real time hemodynamic data, particularly those patients who otherwise may not be afforded a high level of monitoring or those needing hemodynamic monitoring when assessment and treatment are delayed because of inaccessibility of critical care beds or in the cardiac catheterization laboratory. Application of a technology assessment model to impedance cardiography illustrates the utility of this method of hemodynamic monitoring. Careful review and critique of the literature differentiates the available impedance technologies, supports use in areas not traditionally associated with hemodynamic monitoring, such as the home and emergency department, and validates the use of impedance cardiography in place of, or as an indication for, pulmonary artery catheterization. PMID- 10373825 TI - Telemetry monitoring in acute and critical care. AB - With the expansion of higher acuity patients in noncritical care areas, the perceived need for arrhythmia monitoring has also escalated. For institutions pursuing this expansion, many factors must be kept in mind, including patient criteria for telemetry initiation and discontinuation, staff competency of ECG interpretation, safety, technology required, usability, and cost effectiveness. All of these issues must be addressed according to the individual institution's needs and the needs of the patient populations they serve. PMID- 10373826 TI - Pulse oximetry. AB - Pulse oximetry is one of the most commonly applied technologies in acute and critical care. It has the potential to continuously monitor pulmonary function, avoid unnecessary blood gases, and alert clinicians to hypoxemic events that are not readily apparent by physical assessment. Due to these advantages, pulse oximetry has a firm place in health care. Unfortunately, pulse oximetry will not usually have a major impact on reducing hospital resources. In addition, oximetry has the potential to be misused owing to its widespread application. In order to obtain the maximum benefits from this technology, clinicians must be educated about the strengths and limitations of oximetry. If this education effectively changes clinicians' behavior, pulse oximetry will provide an excellent clinical advantage in patient assessment as well as moderate cost benefits. PMID- 10373827 TI - Point of care testing in critical care. AB - POCT is rapidly expanding in today's critical care areas. Nurses need to be involved in the implementation and evaluation process of POCT at every step. Each institution must determine which bedside tests are indicated based on an in-depth analysis of test accuracy, positive clinical impact, and cost-benefit ratio. Generally speaking, the progress in technology over the last two decades has resulted in highly accurate tests used for POCT. However, accuracy of the test is contingent on correct calibration and correct usage by the test performer. Reduced TAT, particularly therapeutic time, is the major advantage to POCT. The clinician identifies the need for a blood analysis and within seconds to minutes has a measurement upon which to change or implement an intervention. For the critically ill patient this can potentially save lives and allow for rapid titration of medications or mechanical ventilation, as well as decrease intubation times and ICU length of stay. Reduction in blood loss for the patient is the second major advantage of POCT. POCT requires 2 drops of blood for analysis versus 3 mL or greater for a test sent to the laboratory for analysis. The cost-benefit examination needs to occur from many views. The cost of education, supplies, and personnel of POCT versus laboratory testing are a few key aspects. However, because different testing techniques exist, it is difficult to do absolute cost comparisons. In addition, cost savings of faster ventilator weaning or decreased ICU length of stay are also important factors to include. Lastly, costs are not just financial costs. In addition, clinicians should examine the cost to patients regarding comfort and quicker discharge. These are quality indicators from the patient's perspective. POCT offers many advantages, but surrounding the implementation of this technology is a multitude of questions that each institution must answer prior to undertaking a POCT program. PMID- 10373828 TI - Outlier management. Influencing the highest resource-consuming areas in acute and critical care. AB - Outliers account for a large amount of technology utilization and resources consumed in acute care, despite accounting for only a small portion of the total patient population. Most current efforts to reduce costs, such as re-engineering and downsizing, are nonspecific methods of controlling costs. Focusing efforts in high-cost areas, such as outlier management, is much more likely to improve patient care and improve the use of technology while achieving real advances in cost control. Most hospitals will have to build an infrastructure to support outlier management, which includes developing clinical staff. These processes will take time and careful planning, but they are essential for the effective management of technology utilization and outliers. The failure to employ focused efforts like outlier management will result in the superficial treatment of high costs in acute care. The benefit to employing these methods leads to the best use of technology and the improved management of a difficult patient population. PMID- 10373829 TI - The world of nurse anesthesia. PMID- 10373830 TI - Complementary therapies. PMID- 10373831 TI - Reiki therapy--a tool for wellness. PMID- 10373832 TI - Healing touch--an energy-based approach to healing. PMID- 10373833 TI - Alternative & complementary therapies--an overview for nursing students. PMID- 10373834 TI - Distance education--is it right for you? PMID- 10373835 TI - Associate degree nursing education today--myths and realities. PMID- 10373836 TI - Questions & answers about diploma programs. PMID- 10373837 TI - When the heart is home. PMID- 10373838 TI - Synthesizing philosophy, theory, and research in holistic nursing. PMID- 10373839 TI - The science and art of aromatherapy. AB - Essential oils have been used for thousands of years. Hippocrates claimed that the way to health was through aromatic baths and massages. Much anecdotal evidence exists regarding aromatherapy's positive effects on recipients. However, well-designed research trials are sparse. Certainly much controversy exists regarding the appropriate way to conduct research on holistic therapies. Can their effects be broken down and studied without contradicting the central premise on which holism is based? The purpose of this article is to review the current state of the science of aromatherapy and to propose future research. The author also offers guidelines for safe aromatherapy practice while awaiting future research on its clinical efficacy. PMID- 10373840 TI - Critical analysis of spirituality and its empirical indicators. Prayer and meaning in life. AB - Spirituality for holistic nursing is defined after a critical analysis of the literature. Spirituality is defined as the experiences and expressions of one's spirit in a unique and dynamic process reflecting faith in God or a supreme being; a connectedness with oneself, others, nature or God; and an integration of all human dimensions. Prayer and meaning in life are described as empirical indicators for appraising spirituality. Prayer is an indicator of the defining attribute of connectedness with God, and meaning in life is an outcome of spirituality. PMID- 10373841 TI - Spirituality of patients recovering from an acute myocardial infarction. A grounded theory study. AB - The purposes of this study were to discover what spirituality means to patients recovering from an acute myocardial infarction and to identify patients' perceptions of how spirituality influences recovery. Using the Glaserian method, spiritual concepts were used for theory building. Spirituality was described as a life-giving force nurtured by receiving presence of the divine, family, friends, health care providers, and creation (core category). Supporting categories were: developing faith, discovering meaning and purpose, and giving the gift of self. Five phases to discovering meaning and purpose were: (a) facing mortality, (b) releasing fear and turmoil, (c) identifying and making lifestyle changes, (d) seeking divine purpose, and (e) making meaning in daily life. Spirituality influenced recovery by providing the participants with inner strength, comfort, peace, wellness, wholeness, and enhanced coping. This substantive theory can be used by holistic nurses to facilitate spirituality in patients recovering from an acute myocardial infarction. PMID- 10373842 TI - Nursing presence. As real as a milky way bar. AB - Data sets from three individual studies on nursing judgment were reviewed from a wider perspective. This yielded meanings and phenomena not readily identified in the individual studies, and it was tentatively labeled presence. A hermeneutic study using 10 transcripts from each data set asked: What are the common features of the context of nursing judgment? and, What are the features of the nurses' connection with the patient that contribute to nursing judgment? The analysis yielded six features of nursing presence: uniqueness, connecting with the patient's experience, sensing, going beyond the scientific data, knowing (what will work and when to act), and being with the patient. These features of nursing presence are logical distinctions and serve as ways to grasp the idea of nursing presence. PMID- 10373843 TI - The willingness of family members of critically ill adults to learn the coping technique of imagery. AB - Critical care nurses face the challenge of helping anxious families cope with the critical illness of one of their members. The purposes of this exploratory descriptive pilot study were to identify whether adult family members of surgical intensive care unit (SICU) patients were willing and able to learn imagery during the time of their family member's critical illness, and which factors facilitate the learning. Of the 139 persons invited, 26 (18.7%) indicated willingness to participate, but only 10 participants completed both learning sessions. All 10 subjects achieved increased relaxation with the imaging. Facilitators of their learning were the quiet environment, the investigator's voice, and the breathing aspect of the technique. Those who participated found imagery beneficial, but further study is needed regarding the basis of both willingness and reluctance to learn relaxation techniques such as imagery to assist with coping during the SICU vigil. PMID- 10373844 TI - An integrative model for holistic community health nursing. AB - Health and nursing are shifting from acute care to community settings, causing confusion for clients and professionals. Although the holistic nursing perspective has improved care to human beings in interaction with their environment and has moved nursing away from pathology-focused care, the community as the focus of care has not been addressed. This article presents an integrated model of community health that expands holistic nursing to the community as client. The model clarifies nursing care for the individual, family, aggregate, and community. One can focus on any one level, with the awareness that each is part of a unified whole. Holistic community nursing completes the circle of care by moving beyond the particular part to focus on the greatest health for the community. The intent is to help nurses describe their unique areas of expertise within the complex community system and to establish a basis for collaboration and partnership. PMID- 10373845 TI - Primary care for the 21st century. PMID- 10373846 TI - Doctor-nurse substitution: the workforce equation. AB - AIM: This paper examines the historical background and context to the doctor nurse substitution debate, and then addresses the sufficiency assumptions inherent in the new nursing roles. BACKGROUND: The NHS Executive considers 'new nursing roles' as a means of substituting part of the doctors' skills. Whilst the literature abounds with professional debate related to the desirability of nurses extending their roles, the underlying assumption of a sufficiency of skilled nurses is not considered. METHODS: The NHS hospital workforce data for the year 1994/95 were analysed and the changes in the overall numbers of doctors and nurses available for work were calculated as the doctors' hours were progressively reduced. FINDINGS: The changes in skill mix were compared; firstly, as a result of the estimated potential reductions in nurses available to undertake the nursing function as movements up the nursing skills spectrum occurs, and secondly, as a result of the alteration in the balance of available skilled staff. CONCLUSION: The policy assumption that suggests that a sufficiency of nurses is available for doctor substitution, whilst still allowing the nursing element to function may be false. PMID- 10373847 TI - Clinical nursing supervision in the workplace--effects on moral stress and job satisfaction. AB - AIM: To investigate nurses' satisfaction with their work environment and moral stress levels as effects of systematic clinical nursing supervision. BACKGROUND: Nurses have identified high workload, low influence over work assignment, limited avenues for skills development and diminishing support from supervisors as sources of considerable tension resulting in deterioration of work conditions and decreased job satisfaction. METHODS: This study is a descriptive-correlational study. Data were analysed by means of descriptive statistics. RESULTS: The major result indicates moral stress in the workplace. It was found that a significant relationship existed between moral sensitivity and systematic nursing clinical supervision. CONCLUSION: The results point to the need to support nurses in developing personal qualities, integrated knowledge and self-awareness, which is in line with the effects of clinical nursing supervision reported in other studies. PMID- 10373848 TI - Developing an instrument to measure patient satisfaction with nursing care in Greece. AB - AIM: The main purpose of this study was to develop a reliable and valid instrument to measure patient satisfaction with nursing care. BACKGROUND: The interest in patient satisfaction is intense but there is an absence of instruments with proper psychometric properties. METHODS: A methodological exploratory design was employed with three phases: content development and critique, pilot study (N = 15) and final tryout (I = 103). FINDINGS: The reliability coefficient for the whole scale was high (alpha = 0.94). An exploratory factor analysis revealed six factors, explaining 68.8% of the variability. The first three factors referred to interpersonal relationships and available time, technical competence and response, and information. The qualitative data facilitated the interpretation of the quantitative data, increased the validity of the scale and gave useful information for improvements. CONCLUSIONS: Overall, the psychometric properties of the instruments were satisfactory but there is a need for continuous evaluation and verification of other studies. PMID- 10373849 TI - Management from four different perspectives. AB - AIM: This study intended to illuminate nursing management in a developing organization from the perspectives of nurse managers, chief physicians, hospital directors and politicians. BACKGROUND: Increased responsibility in a changing health care system makes it important that nurses occupy advantageous positions so that they may safeguard and facilitate the development of their core area. METHODS: Open-ended taped interviews were conducted with 15 nurse managers, 11 chief physicians, three hospital directors and three politicians from three Swedish cities. FINDINGS: The main theme found was power and three dependent themes were identified as; power within activities, being in power and freedom to act. CONCLUSION: The four professional groups related their opinion of nursing management to the needs and interests of their own group. Acceptance or nonacceptance between the groups was discussed in terms of the concept of knowledge, which was interpreted as being a more acceptable topic for discussion than the phenomenon of power. PMID- 10373850 TI - Use of reflective practice in introducing change on the management of pain in a paediatric setting. AB - AIM: The paper describes the application of a reflective model devised by the author, in the management of paediatric pain within a district hospital. BACKGROUND: The issue of bridging the theory-practice gap in nursing has encapsulated the workings of both nurse practitioners and theorists for many years, not least in the management of paediatric pain. METHODS: The reflective model, divided into seven stages, demonstrates the way in which reflective practice and management of change can be integrated. FINDINGS: Applying this model highlighted nurses' concern themselves with a theory-practice gap, but was also evident of a theory (practice knowledge)-theory (official scientific knowledge) gap that needs to be acknowledged. CONCLUSION: It is predicted that the integration of reflective practice with management of change will be useful to nurse practitioners and theorists alike who have sought to address the issue of a theory-practice gap in nursing by embracing the concept of reflective practice. PMID- 10373851 TI - Primary care groups. PMID- 10373852 TI - Perinatal computerized patient record and archiving systems: pitfalls and enhancements for implementing a successful computerized medical record. AB - Interest in purchasing and installing a perinatal computerized patient record (CPR) and archiving system is growing in the United States as a result of increased patient satisfaction demands, cost containment, and quality improvement. Perinatal nurses are commonly charged with researching available computer software and hardware, making purchasing decisions, developing menus and forms, orienting users, and maintaining and upgrading systems. The decision to chart and archive by computer as well as installation and maintenance issues mandate that nurses increase their computer-related knowledge. The article reviews information related to CPR capabilities and rationales for purchase decisions, implementation and staff development issues, ergonomic and maintenance considerations, and realistic expectations of a CPR to provide perinatal nurses who are involved in purchasing, implementing, and maintaining these systems with a timely understanding of important elements that they need to know to make this effort successful. PMID- 10373853 TI - World Wide Web resources for perinatal nursing. AB - The article provides practical guidelines on how to access and search for information resources on topics related to perinatal nursing on the World Wide Web. Categorized listings of key information sites include major relevant databases and bibliographies, electronic journals, sites relevant to perinatal nursing, and community health resources. The Web provides an excellent information resource for health care professionals and the public to gain practical clinical knowledge and to access research resources. Internet technology has altered the way data are perceived and presented because of the speed with which complex data manipulations are now possible and on account of the vast quantities of data involved. PMID- 10373854 TI - Internet communication and discussion lists for perinatal nurses. AB - The article describes how electronic connectivity facilitates professional communication. A brief background in computer communication networks, telecommunications and Internet access, email, and electronic discussion lists prepares the reader to participate in this technology. Directions for subscribing to an online discussion list and guidelines for professional behavior on the Internet are outlined. The article concludes with a description of the Perinatal Nursing Discussion List, an established networking tool. Printed and electronic resources for locating additional lists and a glossary of online abbreviations and terms are included. PMID- 10373855 TI - Internet resources for neonatal nurses. AB - The availability of health information over the Internet has exploded in recent years. Nurses can use the Internet to access information to support professional and clinical interests and join in dialogues with colleagues around the world. The Internet can also be used to identify resources for clients and their families. As with any other information resource, Internet sites must be evaluated for accuracy, currency, and objectivity. The article describes examples of Internet resources and discussion forums for neonatal nurses, use of search engines to find and retrieve information about specific topics, and evaluation of World Wide Web sites. PMID- 10373856 TI - Necrotizing enterocolitis. AB - Necrotizing enterocolitis (NEC) is the most serious and frequently acquired gastrointestinal disorder in neonates. The pathogenesis of NEC is unknown, but it may result from a disturbance of the delicate balance among gastrointestinal perfusion, enteric organisms, and enteral feeding. Risk factors for NEC include prematurity, hypoxic-ischemic insult, and formula or breast milk feedings. The clinical spectrum of NEC is multifactoral and ranges from temperature instability, apnea, lethargy, abdominal distention, bilious residuals, and guaiac positive stools to septic shock, disseminated intravascular coagulation, and death. Medical management is usually adequate treatment for NEC. Surgical treatment is considered if medical management is inadequate to control the spread of the disease. Health care team members must be constantly alert to the presentation of NEC. Expeditious treatment will positively influence the outcome of the disease. PMID- 10373857 TI - Critical congenital cardiac defects in the newborn. AB - Recognizing cyanotic and other critical congenital cardiac defects and providing appropriate supportive care before transport to a cardiac center or unit are crucial in promoting the survival and positive outcome of affected newborns. The article discusses basic pathophysiology, signs, symptoms, and supportive care required for the following defects: transposition of the great arteries, tetralogy of Fallot, tricuspid atresia, truncus arteriosus, total anomalous pulmonary venous return, hypoplastic left heart syndrome, critical aortic stenosis, coarctation of the aorta, and pulmonary atresia with intact ventricular septum. Also discussed are the normal transition from fetal to neonatal circulation, diagnosis, preparation for transport, and treatment of these defects. The information is designed for health care providers in a number of settings. PMID- 10373858 TI - President's message: WOC(ET) nursing and case management. PMID- 10373859 TI - From clinical expert to clinical investigator. PMID- 10373860 TI - Reviewing and understanding research reports. PMID- 10373861 TI - Adult survivors of childhood sexual abuse as patients: two case studies. AB - Adult survivors of childhood sexual abuse comprise a high percentage of the patients seen in gastrointestinal and genitourinary clinics and are commonly found among the patient population treated by WOC nurses. The physical and emotional consequences of sexual abuse may permeate the survivor's life, but rise to the forefront only with the additional stress of an ostomy or urinary diversion. Two case studies are described involving women the authors encountered in their practices. PMID- 10373862 TI - Use of hyperbaric oxygen and negative pressure therapy in the multidisciplinary care of a patient with nonhealing wounds. AB - The case of a 55-year-old woman with nonhealing wounds located on the sternum, abdomen, and lower left extremities is described. The wounds were related to surgical incisions from coronary artery bypass grafting and were complicated by respiratory insufficiency, diabetes mellitus, and infection. This article presents a brief overview of the collaborative care provided in this case and a pictorial review of this patient's wounds during a 4-month period. PMID- 10373863 TI - The effect of ostomy surgery between the ages of 6 and 12 years on psychosocial development during childhood, adolescence, and young adulthood. AB - OBJECTIVE: To investigate the effect of ostomy surgery performed between the ages of 6 and 12 years on psychosocial development during subsequent childhood, adolescence, and young adulthood. DESIGN: Ethnographic. SETTING AND SUBJECTS: Six women and 4 men (mean age 30.7 years) who responded to a notice in a chapter newsletter of the United Ostomy Association. METHODS: Spradley's Ethnographic interview format was used for this study. One interview was conducted with each informant over a 4-week period, and a second, more detailed interview was conducted with 2 subjects who were identified as key informants. The interviews were audiotaped and transcribed, and the transcripts were analyzed to detect shared patterns in the informants' language. INSTRUMENTS: The Ethnograph computer software program was used as an aid in analyzing the interview transcripts. MAIN OUTCOME MEASURES: The informants' subjective experiences during childhood, adolescence, and young adulthood. RESULTS: Nine of the 10 informants adjusted well in the first years after surgery. Key factors in good adjustment were support from family and a perception of "normalcy," including managing one's own ostomy care. All informants reported that their ostomy had a negative impact on their lives during adolescence and that they would have appreciated contact with other teens facing the same dilemma. The age at which an informant underwent ostomy surgery did not influence the difficulties reported during adolescence. CONCLUSION: Ostomy surgery performed between the ages of 6 and 12 years can have long-term effects on psychosocial development. Nurses should promote normalization, teach self-care of the ostomy as soon as possible after surgery, and refer children and parents to mutual support groups as appropriate. PMID- 10373864 TI - A review of the anatomy of the male continence mechanism and the cause of urinary incontinence after prostatectomy. AB - Radical prostatectomy was first described by Dr. Hugh Hampton Young in 1905 as a treatment for prostate cancer. Since that time, urinary incontinence has been reported as a significant postsurgical problem. With the expanding interest in continence therapy and an increase in the number of men undergoing prostate cancer surgery, there is a concomitant need for detailed consideration of the cause of postprostatectomy incontinence. Urinary leakage after radical prostatectomy is not, as traditionally thought, a simple case of stress urinary incontinence. Instead, it represents a complex, multifactorial problem that continues to challenge practitioners and researchers alike. An overview of the anatomy of the male continence mechanism is provided, followed by a discussion of the cause and risk factors implicated in postprostatectomy incontinence and suggestions for further research. PMID- 10373865 TI - A comparison of midnight versus early morning removal of urinary catheters after transurethral resection of the prostate. AB - PURPOSE: This article describes a study that compares the outcomes of midnight versus early morning urethral catheter removal after transurethral resection of the prostate. SUBJECTS AND SETTING/METHODS: The research setting was a large, metropolitan hospital in Sydney, Australia. Forty-eight patients who had undergone transurethral resection of the prostate were randomly assigned to either group A, catheter removal at 2400 hours (n = 20), or group B, catheter removal at 0600 hours (n = 28). MAIN OUTCOME MEASURES: Data collected included time to first void, volume of first void, time between catheter removal and discharge from hospital, weight of prostatic resection, and tissue pathology. RESULTS: There was no significant difference between the 2 groups with respect to pathology, weight of prostatic resection, mean volume of first void, or time to first void after catheter removal. There was a significant difference in the time between catheter removal and discharge from hospital. Eighty-five percent of those having catheters removed at 2400 hours were discharged on the same day as catheter removal, as compared with 65% of those who underwent catheter removal at 0600 hours (chi 2 = 12.684; P < 0.005). CONCLUSION: After transurethral resection of the prostate, removal of the urethral catheter at 2400 hours reduced the length of hospital stay, but did not significantly affect the time to first void or the volume of the first void. PMID- 10373866 TI - Treatment of painful lower extremity ulcers in a patient with sickle cell disease. PMID- 10373867 TI - Development of evidence-based guidelines in midwifery and gynaecology nursing. AB - OBJECTIVE: To develop an effective and efficient method for basing nursing practice on research evidence. SETTING: The Royal Women's Hospital, Brisbane, Australia. METHOD: Nurses and midwives from various clinical areas were invited to participate in an evidence-based practice project. Standard procedures for retrieving relevant articles and evaluating their quality were observed. Where possible, raw data from studies with similar methods were summarised using appropriate statistical tests. FINDINGS: Several guidelines have been developed, staff involved with the project have become 'research literate' and the project is contributing to the hospital-wide quality improvement activities. CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: It is possible to translate research findings into practice when small groups use systematic reviews to develop practice guidelines. PMID- 10373868 TI - Swedish fathers' involvement in and experiences of childbirth preparation and childbirth. AB - OBJECTIVE: To discover the expectations and experiences of childbirth preparation and childbirth of Swedish men in order to contribute to a basis of reflections in the midwifery profession. DESIGN: Three tape-recorded interviews were performed: before and after childbirth preparation, and between one and three weeks after the baby was born. SETTING: Swedish maternity care. PARTICIPANTS: Eleven men who participated with their partners in antenatal classes. MEASUREMENTS AND FINDINGS: The interviews were analysed in several steps and included co-assessments by co workers. Finally, an interpretation based on the concept 'vital involvement' was undertaken. Indications of vital involvement as well as various levels of involvement or distance were found. The participation in childbirth was more demanding than expected for the eleven men. They felt unprepared for an unpredictable process, the experience of time and pain, the woman's action, and their own reactions. The men who were regarded by the authors as vitally involved seemed to manage overwhelming feelings of helplessness during childbirth, to support the women, and experience the meeting with the baby positively. KEY CONCLUSION: It seems important for midwives to meet men individually, design childbirth preparation from men's perspective, follow up interpretations of the content, discuss expectations with regard to the men's role, and assess their experiences during the birth process. PMID- 10373869 TI - Prenatal screening for Down syndrome: a dilemma for the unsupported midwife? AB - OBJECTIVE: To explore the personal and professional concerns of midwives in relation to their experiences with women undergoing serum screening for Down syndrome. DESIGN: Semi-structured interviews. SETTING: A consultant-led maternity unit in the south of England. PARTICIPANTS: Ten midwives based in areas which most commonly support women undergoing this test. METHODS: Interviews were recorded and transcribed and a grounded theory approach involving coding was used to identify categories which were analysed. FINDINGS: Themes identified were: education needs on introduction of the test and in relation to current research about the test; strategies for explaining the concept of 'risk'; personal and professional conflict in relation to the dilemmas raised. KEY CONCLUSIONS: Accurate information (including psychosocial aspects) should be available to midwives prior to the introduction of potentially complex investigations. Continuing education should prepare midwives to meet the challenges related to prenatal screening for Down syndrome. Some strategies which could be considered include the use of reflection sheets, prompt cards and case studies or vignettes. Interprofessional education could provide the opportunity to increase understanding of individual roles and the conflicts experienced. PMID- 10373870 TI - Mixed messages about the meanings of breast-feeding representations in the Australian press and popular magazines. AB - The popular press is an influential medium for the communication of messages and meanings about health and lifestyle issues (Lupton 1993). Through language and images, the print media present selected phenomena, events and issues to readers. The choice and connections between works used can impress upon the reader specific images of the world and the attitudes toward the presented issues and ideas (Nunan 1993). Discourse analysis is used in this study to examine representations of breast feeding in articles published in the Australian press and popular magazines over a six-month period. Discourse analysis is a method of inquiry that focuses on sociocultural and political contexts in which communication occurs (Lupton 1992). Discourses revealed mixed messages and meanings. Breast feeding was seen as natural and the best way of feeding but also as problematic in practice. Dominant ideologies of power and persuasion were also evident. The media portrayed predominantly negative views about breast feeding. Such discourses may influence decisions to breast feed and have wider implications for midwives in their roles as supporters and educators of women and their families. PMID- 10373871 TI - Birthing at home: the resolution of expectations. AB - OBJECTIVE: To describe the experience of ten couples who have had a home birth in Western Australia. DESIGN, SETTING AND PARTICIPANTS: Using a phenomenological approach, ten parent couples were interviewed and three home-birth videos observed. Of the ten couples, four discussed their first child's home birth. The remaining six couples had three or four children who had been born at home. FINDINGS: The essence of these parents' experiences of home birth was gained through identifying significant statements from transcripts and field notes and clustering these into the four themes of 'constructing the environment'; 'assuming control'; 'birthing'; and 'resolving expectations'. The first two themes were presented in a previous paper (Morison et al 1998). The latter two themes are now presented. The theme 'birthing' was where parents elaborated on their birth beliefs, discussed the actual birth and shared aspects of the relationship between the couple and the midwife. 'Resolving expectations' concerned the process of parents forming expectations, experiencing the reality of birth and then evaluating whether expectations were met. KEY CONCLUSIONS: The development of a supportive relationship between couples and their midwife was essential during this transition to parenthood. Resolving expectations was an essential process that the parents undertook to clarify the meaning of their birth experience, and thereby acknowledge its uniqueness. IMPLICATIONS FOR PRACTICE: The findings are important to midwives' practice as they reveal the value clients place on a shared philosophy about birth. Midwives, in any setting, can reflect on their own birthing beliefs and determine their compatibility with their clients' beliefs. PMID- 10373872 TI - A prospective study of women's views of factors contributing to a positive birth experience. AB - OBJECTIVE: To explore the aspects of a woman's childbirth experience which she perceived as being important. DESIGN: As part of a large randomised trial, which assessed the timing of intervention in prolonged labour, women's views were explored using a specifically-designed questionnaire. The questionnaire, which was administered on the second postnatal day, incorporated a rating scale followed by an open question. The responses to the open question are presented in this paper. SETTING: Regional teaching hospital in the north west of England. SAMPLE: 615 primigravid women received a copy of the questionnaire. Of the 519 women who returned the questionnaire, 412 women answered the relevant section, the findings of which are presented in this paper. ANALYSIS: The responses to the open-ended question were analysed by the generation of themes from the most frequently occurring responses. MAIN FINDINGS: The main themes which emerged were support, information, intervention, decision making, control, pain relief and trial participation. KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: Most women are able to identify important contributors to a positive intrapartum experience. Midwives have an important role in identifying these contributors and supporting women to fulfil their individual needs. PMID- 10373873 TI - The experiences of postpartum hospital stay and returning home among Thai mothers in Australia. AB - OBJECTIVE: To describe the experience of postpartum care among Thai women in Melbourne, Australia. DESIGN: Ethnographic interviews and participant observation with women in relation to postpartum care and practices. SETTING: Melbourne Metropolitan Area, Victoria, Australia. PARTICIPANTS: 26 Thai born women who gave birth in Australia. FINDINGS: The Thai women had varying views about the length of time they should spend in hospital and the care they received. Ten of the twelve women who had had a caesarean birth stayed in hospital for six or more days, consistent with the hospital practice. However, most of those who had had a vaginal delivery opted to go home earlier than the standard hospital practice of four days stay. This was because they were unhappy about specific hospital practices, the hospital environment, or because there are several Thai confinement customs, which, traditionally, a new mother must observe in order to maintain good health and avoid future ill health and which they were not able to follow in hospital. Nevertheless, most women were satisfied with their postpartum care. Most women were aware of the Thai cultural beliefs and practices. However, they showed varying ways of coping with the hospital environment in relation to their varying social situations. IMPLICATIONS FOR PRACTICE: Thai women are diverse in their needs, perceptions and experience of postpartum care. Therefore, it is appropriate neither to stereotype all Thai women as requiring to follow traditional confinement practices nor to require them to adjust to standard hospital practices. Rather an environment of caring concern whereby each woman's individual needs can be solicited, understood and, where possible, attended to as required. The challenge is in achieving this. PMID- 10373874 TI - Make certification a consumer concern. PMID- 10373875 TI - Who's liable in equipment cases? AB - An increase in the number of equipment-related liability cases underscores providers' and manufacturers' joint responsibility to ensure patient safety. PMID- 10373877 TI - Manage moonlighting. PMID- 10373876 TI - Managed care's move to rural areas. AB - Medicare, Medicaid, and urban-based commercial health plans are attempting to increase managed care's presence in the nation's rural areas, prompting rural based providers to sponsor their own managed-care plans. PMID- 10373878 TI - Duties and direction for managing contract nurses. AB - The author explains how the human resource chapter affects nurse managers' responsibilities toward contracted staff and lists ways to organize orientation and verify competence. PMID- 10373879 TI - Nursing's role in Y2K planning. AB - Why haven't more nurses fulfilled their role in Y2K planning? Nurses need to apply their skills and expertise to solving often overlooked problems such as point-of-service applications, transactions with business partners, and contingency planning. PMID- 10373880 TI - Take teamwork to new heights. AB - Teamwork follows the dynamic path of group development. The authors evaluate their survey of teamwork in 14 ICUs and explain how nurse managers can evaluate and increase teamwork with a unit assessment process. PMID- 10373881 TI - How to merge telemedicine with traditional clinical practice. AB - Using telemedicine to care for homebound patients increases patients' access to care, reduces resource consumption, and improves use of scarce health professionals. PMID- 10373882 TI - Strategies for reporting to a nonnurse vice-president. AB - Organizations reorganize in very dramatic ways. This article describes the challenges of reporting to someone from a different profession. PMID- 10373883 TI - Six stepping stones to better management. AB - Clinical pathways can help nurse managers develop clinical staff, guide quality improvement, enhance interdisciplinary practice, standardize care delivery, control the budget, and increase patient satisfaction. PMID- 10373884 TI - Critical thinking, critical practice. AB - Nursing leaders in critical care can use strategies such as questioning, clinical scenarios, conferences, and context-dependent test items to assess and improve their staff's critical thinking skills. PMID- 10373885 TI - Anatomy of a disease management program. AB - Learn the strategies and planning behind disease management programs. Initiation and success depend on nurse leaders, case managers, and information technology. PMID- 10373886 TI - Sick-child daycare promotes healing and staffing. AB - Keeping working parents working when their children are mildly ill protects workplace productivity. This article presents a successful model for a medical center's sick-child day-care facility. PMID- 10373887 TI - Empowering lifelong learners. AB - How can nurse managers provide staff with equitable, economical opportunities for professional development education? One hospital's point accrual system makes nurses more active in their own professional development. PMID- 10373888 TI - The freedom to roam. AB - Wireless devices' many applications are changing the way nurses work. Read how to select wireless solutions that enhance your performance. PMID- 10373889 TI - Tangled up in "nots". PMID- 10373890 TI - The termination process: strategies for nurse managers. PMID- 10373892 TI - Ask AONE's experts ... about how to incorporate case managers' costs. PMID- 10373893 TI - Nursing has a value that is hard to measure. PMID- 10373891 TI - It's not what you say, but how you say it. PMID- 10373894 TI - We won't take this lying down. PMID- 10373895 TI - Nurses have a duty to their patients to help them exercise their voting rights. PMID- 10373896 TI - Pressing charges. PMID- 10373898 TI - De Witt to succeed. Interview by Adrian O'Dowd. PMID- 10373897 TI - Drama out of a crisis. Interview by Rebecca Coombes. PMID- 10373899 TI - What are Beacons? PMID- 10373900 TI - The debate over uniforms. PMID- 10373901 TI - The way long-term care is delivered. PMID- 10373902 TI - Time for nurses to find their voice. PMID- 10373903 TI - Why I gave up the high life. PMID- 10373904 TI - Face to face. Interview by Eileen Fursland. PMID- 10373905 TI - A new take on old problems. PMID- 10373906 TI - To Russia with cling film. PMID- 10373907 TI - Home insurance. PMID- 10373908 TI - So you want to be a ... naturopath. PMID- 10373909 TI - The adoption of a healthy eating programme in Somerset schools. PMID- 10373910 TI - Handling complaints in a constructive way. AB - This article offers advice on dealing with specific complaints and looks at the nurse's role and responsibilities in handling and resolving them. PMID- 10373911 TI - Music therapy for people with life-limiting illness. PMID- 10373913 TI - NT. Good Practice Network. On your marks. PMID- 10373912 TI - Enhancing district nurses' practice. PMID- 10373914 TI - The future beckons. PMID- 10373915 TI - Twelve years of hell. PMID- 10373916 TI - Designer gear. PMID- 10373917 TI - Know how. Managing incontinence. PMID- 10373918 TI - Making contact. PMID- 10373919 TI - Eliminate the negative. PMID- 10373920 TI - Accentuate the positive. PMID- 10373921 TI - Meeting the challenge. PMID- 10373922 TI - Mines of information. PMID- 10373923 TI - Students felt responsibility for patients but were powerless over what was filmed. PMID- 10373924 TI - Scotland the brave. PMID- 10373925 TI - The great divide. PMID- 10373926 TI - Across the border. PMID- 10373927 TI - Healing the divisions. PMID- 10373928 TI - Who cares now in Kosovo? PMID- 10373929 TI - What's HIP? Health improvement programmes. PMID- 10373930 TI - Can patients ever be a priority for managers? PMID- 10373931 TI - Hospitals without patients. PMID- 10373932 TI - Reasons to be cheerful. PMID- 10373933 TI - Pregnant women and diabetes. PMID- 10373935 TI - Wise councils. PMID- 10373934 TI - Colony to community. Interview by James Staples. PMID- 10373936 TI - So you want to be a ... counsellor. PMID- 10373937 TI - The budget. PMID- 10373938 TI - Ship-shape memories. PMID- 10373939 TI - Brevity is an essential skill in keeping useful patient records. PMID- 10373940 TI - Why action is needed to stop spread of hepatitis C. PMID- 10373941 TI - White-knuckle ride to theatre. AB - Preventing or reducing anxiety in surgical patients can lead to better recovery with fewer complications and earlier discharge (Baldwin 1993). There seems to be little evidence, however, that effective nursing measures are taken to reduce anxiety. This article presents a critical review of the literature on preoperative anxiety, and what is being done to alleviate it. It makes recommendations for future practice, emphasising psychological support of the patient and individualised care. PMID- 10373942 TI - NT. Good Practice Network. Networking works. PMID- 10373943 TI - Housebound older people are missing out on diabetes care. PMID- 10373944 TI - NHS nurses could learn a lot from their colleagues in nursing homes. PMID- 10373945 TI - Wish you weren't here. PMID- 10373946 TI - Bravery and bravado. PMID- 10373947 TI - Decent disclosure. PMID- 10373948 TI - What is Sure Start? PMID- 10373949 TI - Unison's health group conference a week away. PMID- 10373950 TI - Community nurses are never alone with a hand-held radio. PMID- 10373951 TI - Nurses are sick of being abused. PMID- 10373952 TI - Goodbye to gagging: know your rights. PMID- 10373953 TI - Open to persuasion. PMID- 10373954 TI - Single care homes. United we fall. PMID- 10373955 TI - Patient view. Making slow progress. PMID- 10373956 TI - Trust watch. Powers of diplomacy. PMID- 10373957 TI - Foreign exchange. PMID- 10373958 TI - So you want to be a ... solicitor. PMID- 10373959 TI - A barter system. PMID- 10373960 TI - Primary health care records: an integrated approach. AB - The extended primary health care team is expected to achieve the government's vision of an integrated service for patients. Key to this vision is an integrated record-keeping system, integrating GP and community nursing functions. PMID- 10373961 TI - The benefits of assertive community treatment. PMID- 10373962 TI - Learning to cope with the fatigue caused by cancer. AB - Nurses working in many different specialties care for people with cancer. Many of these patients are in the palliative phase of their illness: they are no longer looking for a cure but quality of life is of the utmost importance. This article outlines the development of a patient information leaflet on fatigue, one of the most common and debilitating symptoms experienced by this group of people. PMID- 10373963 TI - Can you hear yourself think? PMID- 10373964 TI - One person's racket is another person's rock and roll. Discuss. PMID- 10373965 TI - Reality bites. PMID- 10373966 TI - Changing from person to ... cancer patient. PMID- 10373967 TI - The unbearable lightness of learning. PMID- 10373968 TI - Ward placements can be daunting. PMID- 10373969 TI - The effects of relocation on the elderly. PMID- 10373970 TI - Using the towel bath to give tender care in dementia: a case example. AB - Using the "towel" bath to clean the skin for such residents as Helen results in a reduction of agitation and an increase in comfort as the skin is cleansed. The "towel" bath is a very useful technique which should be incorporated into educational training programs for any care provider who will be working with persons with dementia. It is a procedure that can be adapted to the care of the most challenging resident and will help staff begin to display person-centred care for the resident who is afraid of care procedures and responds negatively as a result. The procedure itself addresses the issues of water temperature and privacy in a way that traditional basin bathing makes difficult. The "towel" bath should be included in the practice repertoire and used on a regular basis by all practitioners who work in this area of geriatric specialty. PMID- 10373971 TI - Euthanasia and the elderly. PMID- 10373972 TI - Caring for the older client on i.v. therapy. PMID- 10373973 TI - Bright light treatment for people with Alzheimer's disease. PMID- 10373974 TI - Cultural diversity: what is it really about? PMID- 10373975 TI - Types of budgets. PMID- 10373976 TI - Cultural diversity as a strength: building symbiotic relationships. PMID- 10373977 TI - Managing cultural diversity: when organizations merge. PMID- 10373978 TI - Transforming cultural chaos. AB - Kotter believes that generating short-term wins is a critical component of the change and transformation process. Our short-term win-wins have allowed us to further collaborate, which consequently has given us more wins. These wins have been communicated. The communication has conveyed a sense of optimism and hope, and has empowered people to view change as a positive challenge. The challenge has become one in which all want to participate. Finally, we are at a point where the end of one way of life is truly the beginning of a new and better way of life. T.S. Eliot was right! PMID- 10373979 TI - Organizational culture in the changing workplace: an employee assistance program perspective. AB - The rapid change experienced in the work world today results in an often dramatic change in the organizational culture. In the process of culture change, employees experience loss of identity and loss of meaning in their work. These losses will manifest themselves in the workplace in the form of withdrawal, isolation, the failure of teamwork, and a high conflict milieu. It is helpful to the effective navigation of the change process to facilitate the grieving of these losses. Only when these losses have been successfully mitigated will employees experience a resurgence of energy and commitment. PMID- 10373980 TI - Cultural chaos in the trenches: staff perspectives. AB - When a hospital is acquired by a large health care organization, there are many challenges, for both the hospital and the organization. When the differences in culture are not addressed effectively by management at the time of acquisition, this can result in cultural chaos for the staff and a lengthened transition time. In this article, a staff member shares the observations, opinions, and feelings she experienced during such a time of cultural chaos between two well-respected organizations. Her opinions and observations are juxtaposed with the lives of two amalgamated fictionalized characters. PMID- 10373981 TI - Change in health care systems: one physician's perspective. PMID- 10373982 TI - Economic climate and cultural diversity: a merging picture. AB - This article explores the merging of a single nonprofit hospital with a nonprofit health care system of hospitals. It also provides the unique financial history of health care reimbursement in California that makes such merges imperative for the success of single hospitals. A unique patient-focused care model, shared governance, and the implications in the financial diversity of merging health care entities also are discussed. PMID- 10373983 TI - Developing a mission, vision, and philosophy in the new context of cultural diversification. AB - This article is about the blending of a mission, vision, and philosophy of care by two systems of health care that are both rich in history and vision. The unique qualities of each hospital are described. The diversified cultures of each organization are discussed in terms of reaching a final decision regarding the joint vision, philosophy of care, and mission of the system that has been redesigned. PMID- 10373985 TI - Marketing culture to the community. AB - When the merging of organizational cultures comes together, a plan must be implemented to market the new organization to the community. Community, in this instance, is defined as the patients, employees, physicians, insurers, and any others who may be affected by a change in the health care system. This article focuses on a merging organizational culture and the marketing of that new culture. PMID- 10373984 TI - Understanding the impact of power in organizations. AB - Although implementation of Shared Governance appears to have failed, it failed primarily on the surface. Many staff nurses actively involved in the Shared Governance movement not only were empowered but were dramatically affected on a professional level. Several council chairpersons were empowered to assume management roles in the transition back to the hierarchial model--a manifestation of their professional growth and development. At the unit level, several units lobbied the new leadership to allow them to continue to do peer review and unit based council management of unit governance issues. Three councils lobbied to continue to do their work, although in a modified role, in the reestablished hierarchial structure. The three remaining councils were those of Practice, Quality, and Research. If nurse leaders at any level within the organization are to guide their departments forward while in the throes of the current chaos in health care, they must develop and use their power bases, both formal and informal, as individuals and then as leaders. Russell Coile identifies the need for more clinical expertise (expert power) on the executive team of health care organizations. He predicts that 50% of the executive team will be nurses and physicians and that only health care executives with an MBA or financial backgrounds, who also have well-developed informal power bases with skills in relationship development, facilitation, and networking, will be part of the new system. Those with a diversified informal power base will be most successful in guiding their organization to its destination. The future for nursing leaders is in the sharing of information; it is about embracing diversity and recognizing the contributions others can make that are refreshingly different; it is also about clearly defining a balance in life, because balanced leaders who have found a way to nurture and meet their own needs are better positioned to do the same for others. Ultimately, understanding the impact of power in an organization, regardless of organizational structure, begins with understanding and defining your own power base. PMID- 10373986 TI - 'Mom, I want to be a nurse!'. PMID- 10373987 TI - Female genital mutilation: perspectives, risks, and complications. AB - Female genital mutilation, traditionally known as female circumcision, is a surgically unnecessary modification of the female genitalia, practiced in nations in Africa, the Arab Peninsula, among some communities in Asia, and among immigrants and refugees from these areas who have settled in other areas. The practice is known across socio-economic classes and among many different ethnic and cultural groups, including Christians, Muslims, Jews, and followers of indigenous African religions. As people from these areas immigrate to North America, health care professionals need to understand the important aspects of this growing problem, including management of complications, cultural attitudes, and sensitivities. PMID- 10373988 TI - Trial of voiding: what's the verdict? AB - Trial of voiding assesses a patient's ability to urinate after removal of an indwelling catheter. An investigation to determine how the procedure was performed at 42 sites nationwide was conducted. The findings are summarized. PMID- 10373989 TI - Electrovaporization of the prostate: initial experiences and nursing management. AB - STUDY DESCRIPTION: Electrocautery power is increasingly used to destroy prostatic tissue as well as to control bleeding during transurethral surgery for benign prostatic hyperplasia. This study summarizes our initial experiences with the electrovaporization procedure and combines these with the published findings of others in order to provide recommendations for the nursing management of this increasingly common urologic surgical procedure. METHODS: A retrospective review of 36 consecutive patients managed by electrovaporization of the prostate was completed. A standardized data collection form was used to evaluate the preoperative preparation, intraoperative experiences, and short-term postoperative responses to the electrovaporization procedure. These data were combined with the published experiences of others in order to provide initial insights into the nursing management of patients undergoing this procedure. RESULTS: Patient preparation was similar to that used for transurethral resection of the prostate. No patients experienced significant intraoperative bleeding. Two subjects (7%) experienced significant but transient hematuria in the postoperative period. Five men (17%) experienced urinary retention requiring catheterization, of these three were caused by blood clots and two were attributable to pre-existing detrusor muscle weakness. Urinary tract infection occurred in eight patients (27%), three of which were accompanied by a fever. All patients reported urethral and suprapubic pain following the procedure. One subject experienced flank pain caused by an acute obstruction of a solitary ureter, and one reported bladder spasms. Significant pain requiring analgesia resolved within 5 days of the procedure in all cases. CONCLUSIONS: The preoperative nursing management of the patient undergoing electrovaporization of prostate tissue is similar to that for transurethral resection of the prostate with the exception of patient teaching. Patient education should emphasize self assessment for urinary retention, hematuria, and urinary tract infection. The postprocedural care initially focuses on monitoring the patient for catheter patency, hematuria, and infection. Patients and significant others should be warned that delayed hematuria and urinary retention may occur days or weeks after electrovaporization of the prostate. During the past decade, the number of alternative procedures for prostate tissue destruction has grown significantly. Each of these procedures offers potential advantages when compared to the classic transurethral resection of the prostate (TURP). While their usefulness in treating obstruction due to benign prostate enlargement has been reasonably well described (McCullough, 1998), the nursing management of patients undergoing many of these procedures has not been adequately studied or described. The purpose of this study was to summarize our initial experiences with the VaporTrode procedure and to better define the nursing management of patients undergoing this increasingly common urologic surgery. PMID- 10373990 TI - Treatments for patients with pelvic pain. AB - An overview of the incidence and types of pelvic pain is presented, followed by some practical information concerning the presentation of patients with pelvic pain. Simple physical therapy techniques that are use in treating patients with pelvic pain are also discussed. PMID- 10373991 TI - Maintenance therapy with bacillus Calmette-Guerin in patients with superficial bladder cancer. AB - The effectiveness of intravesical therapy with bacillus Calmette-Guerin for the treatment of superficial bladder cancer has been well established. However, there is no consensus about the optimum regimen for administering BCG. Maintenance therapy in 28 patients, results, and complications are addressed. PMID- 10373992 TI - Getting ready for certification: pediatric urology for the advanced practice urologic nurse. PMID- 10373993 TI - The evidence for insulin lispro. PMID- 10373994 TI - Adjudicating ethics in research. PMID- 10373995 TI - Adjudicating ethics in research. PMID- 10373996 TI - End-stage renal disease in Canada: prevalence projections to 2005. AB - BACKGROUND: The incidence and prevalence of end-stage renal disease (ESRD) have increased greatly in Canada over the last 2 decades. Because of the high cost of therapy, predicting numbers of patients who will require dialysis and transplantation is necessary for nephrologists and health care planners. METHODS: The authors projected ESRD incidence rates and therapy-specific prevalence by province to the year 2005 using 1981-1996 data obtained from the Canadian Organ Replacement Register. The model incorporated Poisson regression to project incidence rates, and a Markov model for patient follow-up. RESULTS: Continued large increases in ESRD incidence and prevalence were projected, particularly among people with diabetes mellitus. As of Dec. 31, 1996, there were 17,807 patients receiving renal replacement therapy in Canada. This number was projected to climb to 32,952 by the end of 2005, for a relative increase of 85% and a mean annual increase of 5.8%. The increased prevalence was projected to be greatest for peritoneal dialysis (6.0% annually), followed by hemodialysis (5.9%) and functioning kidney transplant (5.7%). The projected annual increases in prevalence by province ranged from 4.4%, in Saskatchewan, to 7.5%, in Alberta. INTERPRETATION: The projected increases are plausible when one considers that the incidence of ESRD per million population in the United States and other countries far exceeds that in Canada. The authors predict a continued and increasing short fall in resources to accommodate the expected increased in ESRD prevalence. PMID- 10373997 TI - Examination for sexual assault: is the documentation of physical injury associated with the laying of charges? A retrospective cohort study. AB - BACKGROUND: Few studies have examined whether there is an association between individual medical findings and legal outcome in cases of sexual assault. This study was undertaken to determine the relation between the extent of documented physical injury and a positive legal outcome in cases of sexual assault and to determine other factors associated with the laying of charges in such cases. METHODS: In this retrospective cohort study, the authors reviewed the charts and medicolegal reports for all cases of sexual assault that were handled by the BC Women's Sexual Assault Service in 1992 for which a police report had been filed. Information on patients' characteristics, the nature of the assault and the extent of injury was extracted from these records. A system for scoring clinical injury was developed by 4 of the physicians at the Sexual Assault Service, and a clinical injury score was assigned for each case by one physician. The relation between the outcome (in terms of whether charges were laid) and the circumstances of the case was examined by logistic regression. RESULTS: A total of 95 cases with complete medical records and information about legal outcome were identified during the 1992 calendar year. After adjustment for income level and the patient's knowledge of the assailant (either as an acquaintance or as his or her partner), the odds ratio (OR) for charge-laying in a sexual assault case with documented moderate to severe injury was 3.33 (95% confidence interval [CI] 1.06 10.42). Socioeconomic status above the group median (defined as annual income greater than $21,893) (OR 3.26, 95% CI 1.09-9.71) and knowledge of the assailant (OR 4.58, 95% CI 1.52-13.79) were also associated with charge-laying. Presence of genital injury per se, age of the patient and detection of sperm by microscopy at the time of examination were not associated with the laying of charges. INTERPRETATION: The results of this study show that the extent of documented injury is associated with the laying of charges in cases of sexual assault. However, many questions remain about the effectiveness of the medical component of gathering such evidence. PMID- 10373998 TI - Non-heart-beating organ donors as a source of kidneys for transplantation: a chart review. AB - BACKGROUND: Organ transplantation is the treatment of choice for patients with end-stage organ failure, but the supply of organs has not increased to meet demand. This study was undertaken to determine the potential for kidney donation from patients with irremediable brain injuries who do not meet the criteria for brain death and who experience cardiopulmonary arrest after withdrawal of ventilatory support (controlled non-heart-beating organ donors). METHODS: The charts of 209 patients who died during 1995 in the Emergency Department and the intensive care unit at the Foothills Hospital in Calgary were reviewed. The records of patients who met the criteria for controlled non-heart-beating organ donation were studied in detail. The main outcome measure was the time from discontinuation of ventilation until cardiopulmonary arrest. RESULTS: Seventeen potential controlled non-heart-beating organ donors were identified. Their mean age was 62 (standard deviation 19) years. Twelve of the patients (71%) had had a cerebrovascular accident, and more than half (10 [59%]) did not meet the criteria for brain death because one or more brain stem reflexes were present. At the time of withdrawal of ventilatory support, the mean serum creatinine level was 71 (29) mumol/L, mean urine output was 214 (178) mL/h, and 9 (53%) patients were receiving inotropic agents. The mean time from withdrawal of ventilatory support to cardiac arrest was 2.3 (5.0) hours; 13 of the 17 patients died within 1 hour, and all but one died within 6 hours. For the year for which charts were reviewed, 33 potential conventional donors (people whose hearts were beating) were identified, of whom 21 (64%) became donors. On the assumption that 40% of the potential controlled non-heart-beating donors would not in fact have been donors (25% because of family refusal and 15% because of nonviability of the organs), there might have been 10 additional donors, which would have increased the supply of cadaveric kidneys for transplantation by 48%. INTERPRETATION: A significant number of viable kidneys could be retrieved and transplanted if eligibility for kidney donation was extended to include controlled non-heart-beating organ donors. PMID- 10373999 TI - Is telephone counselling a useful addition to physician advice and nicotine replacement therapy in helping patients to stop smoking? A randomized controlled trial. AB - BACKGROUND: The authors evaluated the incremental efficacy of telephone counselling by a nurse in addition to physician advice and nicotine replacement therapy in helping patients to stop smoking. METHODS: The trial was conducted at the University of Ottawa Heart Institute. A total of 396 volunteers who smoked 15 or more cigarettes daily were randomly assigned to either of 2 groups: usual care (control group) and usual care plus telephone counselling (intervention group); the groups were stratified by sex and degree of nicotine dependence. Usual care involved the receipt of physician advice on 3 occasions, self-help materials and 12 weeks of nicotine replacement therapy. Telephone counselling was provided by a nurse at 2, 6 and 13 weeks after the target quit date. Point-prevalent quit rates were determined at 52 weeks after the target quit date. RESULTS: The point prevalent quit rates at 52 weeks did not differ significantly between the control and intervention groups (24.1% v. 23.4% respectively). The quit rates did not differ significantly at the secondary measurement points of 4, 12 and 26 weeks. INTERPRETATION: Brief physician assistance, along with nicotine replacement therapy, can help well-motivated smokers to quit. Three additional sessions of telephone counselling by a nurse were ineffective in increasing quit rates. This form of assistance may be useful in the absence of physician advice or when self selected by patients. PMID- 10374000 TI - Rates of adverse events among hospital admissions and day surgeries in Ontario from 1992 to 1997. PMID- 10374001 TI - The price and challenges of extraordinary success: treating end-stage renal failure in the next millennium. PMID- 10374002 TI - Death provides renewed life for some, but ethical hazards for transplant teams. PMID- 10374003 TI - The folly of population screening for type 2 diabetes. PMID- 10374005 TI - Tuberculosis: 6. Extrapulmonary disease. PMID- 10374004 TI - Preventive medicine in people at high risk for chronic disease: the value of identifying and treating diabetes. PMID- 10374006 TI - New virus emerges in Malaysia. PMID- 10374007 TI - Claims to fame. PMID- 10374008 TI - Private company offers hope to cancer patients--for a price. PMID- 10374009 TI - Will voters' response to health care reforms determine the fate of Mike Harris? PMID- 10374010 TI - [The central melanocortin system and its role in energy homeostasis]. AB - Obesity is an important health concern, and the central melanocortin system is likely to play an important role in both normal regulation of energy homeostasis as well as genetic predisposition to obesity. Three different mouse models of obesity have been created by virtue of mutations which alter the function of the melanocortin system, and a rare form of human obesity has been characterized in two families with mutations in the proopiomelanocortin gene [26]. More recently published works suggests an association between common childhood obesity and dominant inheritance of a single null allele of the melanocortin-4 receptor [44, 49]. Experimental work in the rodent suggests that the central melanocortin system may be a fundamental component of the adipostat, the mechanism by which constant energy stores are maintained. This hypothesis is the subject of this review. PMID- 10374011 TI - [The history of science with regard to the thyroid gland (1800-1960)]. AB - This review describes the changing medical perspectives from 1800 to 1960 with regard to the thyroid gland. During the XIXth century, clinical medicine identified the different thyroid diseases: goitrous cretinism (1802), Graves disease (1835-1856) and myxedema (acquired or post-surgery 1888). These clinical entities facilitated the construction of the first complete schema of endocrinology: one gland, one internal secretion and one replacement therapy with an organ extract. With the emergence of biochemistry at the beginning of the XXth century, the first thyroid hormone--thyroxin--was identified. Alternative medical treatments aimed at blocking thyroid secretion, were proposed in 1940 leading to the use of radioiodine and antithyroid compounds which were also used in physiological studies. From 1940 to 1970, the application of radioiodine shed a completely new light on thyroid physiopathology. The main transformations brought about by this tool were the knowledge of radioiodine uptake mechanisms, basis of its therapeutic effect, complete identification of thyroid hormonosynthesis, serum transport of thyroid hormones and thyroid imaging. More recently immunological and molecular paradigms changed the understanding of thyroid diseases. In spite of novel definitions, there was no therapeutic revolution like those introduced by iodine treatment (1820) thyroid surgery (1880), thyroid extract (1893) radioiodine and antithyroid drugs (1940), underlining the actual therapeutic stagnation. CONCLUSION: The prevalence of sub-clinical thyroid dysfunction was 8%. No male subject was affected. The prevalence of nodule was close to 15%. Contrary to the literature, we have not found high incidence of immunological abnormalities in this old population. PMID- 10374012 TI - [Assessment of sub-clinical thyroid disfunction in aged hospitalized patients in Limousin]. AB - The aims of the study were to assess the prevalence of sub-clinical thyroid dysfunction in older patients and to analyze morphological and immunological thyroid abnormalities. SUBJECTS AND METHODS: The effect of aging on thyroid function morphology and immunology was studied in 102 french patients (70 women and 32 men) with a mean age of 81.7 +/- 7.3 years (65-101 years). All patients were hospitalized with different pathological conditions but without any thyroid disease. Patients with treatment or iodine substance which could modify thyroid function were excluded. Serum Free thyroxine (FT4) and TSH levels were detected by RIA. Morphological thyroid study was performed by clinical evaluation and ultrasonography. Two sub-groups of patients were distinguished according to immunological detection tests: In group A (n = 64) immunological parameter was performed by antithyroid microsomal antibodies and in group B (N = 38) by thyroid peroxidase antibodies. RESULTS: We found 4 hypothyroidisms (3.9%), 3 hyperthyroidisms (3%) and 4 marginal isolated high FT4 levels. Elevated levels of FT4 was observed in 3 patients and 1 care of low T4 was described. All patients with abnormal hormonal levels were female. Other patient had normal thyroid function: mean level FT4 was 11.2 +/- 2.2 pg/ml (N: 7-17) and mean level TSH was 1.6 +/- 0.8 mU/l (N: 0.2-4). These values are comparable to those observed in adult populations. Incidence of thyroid autoimmunity was very low (3.9%) in this series compared with previous data. From the morphological analysis, we found 32 morphological abnormalities (31.4%), without strong relation with thyroid dysfunction. Nodule prevalence was near 15% and goiter near 10%. PMID- 10374013 TI - [Hyperthyroidism and diabetes mellitus: analysis of 10 African cases]. AB - This retrospective study of 10 patients with hyperthyroidisma and diabetes mellitus concerned 8 women and 2 men, aged from 15 to 77 years. The two disease developed at the same time in 8 cases. Diabetes mellitus occurred first in 2 cases. Common signs were loss of weight. Hyperthyroidism led to tachycardia at more than 100 bpm. Diarrhea was observed simultaneously in 2 cases and muscular weakness in 5. Goiter was found in 10 cases with a diffuse aspect. Graves' disease was diagnosed with exophthalmia in 9 cases and affected both eyes in 8. Elevated levels of thyroid hormones confirmed diagnosis in 8 cases. Diabetes was insulin-dependent in 3 cases and non-insulin dependent in the 7 others. IDDM patients (2 female and 1 male) were aged 15, 17 and 38. Keto acidosis was the first symptom in all cases. NIDDM patients (6 female and 1 male) were aged between 37 and 77. PMID- 10374014 TI - [Analytical evaluation of third generation TSH assay. Application to the exploration of thyrotropin suppression in differentiated thyroid cancer]. AB - Intravenous TRH (200 micrograms Protireline) injection was performed in 137 patients who had undergone thyroidectomy for differentiated thyroid carcinoma and treated by a suppressive thyroid therapy. Serum thyroid stimulating hormone (TSH) was measured before and 30 minutes after the injection. 143 tests were performed. The TSH method used was a chemiluminometric assay (Chiron Diagnostics). We study the analytical characteristics of the method. The limit of detection was 0.003 mUI/l. Interassay precision showed a functional sensitivity of 0.015 m UI/l with 20% interassay CV in agreement with the third generation criterion. There was a good correlation between TSH values at T0 and T30 minutes. In 143 tests, 40 (28%) showed a basal TSH value less than 0.003 mUI/l; delta TSH (TSH at T30-TSH at T0) was less than 0.015 mUI/l in about 68% of the cases and less than 0.031 mUI/l for the others. In 20% of the tests (29) TSH T0 were between 0.003 and 0.015 mUI/l and delta TSH less than 0.204 mUI/l. Finally the 74 remaining tests with a basal TSH value higher than the functional sensitivity of the method showed delta TSH less than 0.4 mUI/l if the TSH T0 was less than 0.05 mUI/l. The delta TSH remained less than 0.6 mUI/l if the TSH T0 was less than 0.1 mUI/l. Inversely, if TSH T0 was higher than 0.1 mUI/l delta TSH could be higher than 1.0 mUI/l. Third generation TSH assay allows us to abandon TRH test for the follow-up of thyroidectomized patients with suppressive thyroid hormonotherapy in so far as TSH assay is well established. We conclude that a basal TSH less than the functional sensitivity (0.015 mUI/l) can predict a TSH response to TRH stimulation test less than 0.2 mUI/l. PMID- 10374015 TI - [Intrathyroid metastasis of kidney cancer. A rare case and diagnostic trap]. AB - We report the discovery of a unique renal-cell carcinoma operated 9 years earlier on a thyroidectomy operative specimen. Intra-thyroid metastasis from kidney cancer is uncommon. The average delay to discovery as a painless thyroid nodule is estimated around 6 years following nephrectomy. Pre-operative fine needle aspiration and cytology provide the diagnosis. Prognosis of unique thyroid metastasis of kidney cancer is good despite the distant spread. Surgery is the most common therapeutic attitude. PMID- 10374016 TI - [Thyroid hormone determination at the Public Assistance Hospital of Paris: ordering, costs and opinions of physicians qualified in endocrinology]. AB - OBJECTIVES: To determine the distribution of orders for hormonal tests assessing thyroid function in a hospital setting. To collect the opinion of physicians specialized in endocrinology concerning free triodothyronine (FT3) assessment. METHODS: Using a cross-sectional survey numbers of free thyroxine (FT4), total T4 (TT4), FT3, total T3 (TT3), and TSH tests were collected from the heads of laboratory assessing thyroid function in June 95 at the Assistance Publique Hopitaux de Paris (AP-HP). Cost for these tests was estimated. The physicians of the AP-HP specialized in endocrinology were asked through a questionnaire for circumstances justifying FT3 test ordering. RESULTS: Twenty-eight laboratories (93%) responded: 28455 measurements (TSH: 43%, FT4: 33%, TT4: 2%, FT3: 20%, TT3: 2%) were performed and were valued at 3.4 million French Francs. Proportions of T4 (36%) and T3 (20%) tests were lower in hospitals with an inpatient department of endocrinology than in hospitals with an outpatient clinic with specialists in endocrinology (T4: 36%; T3: 27%) or with no endocrinology unit (T4: 33%, T3: 27%); proportion of TSH tests was higher in hospitals with an inpatient endocrinology unit (respectively 44%, 40%, 32%). Forty-two endocrinologists (76%) from 21 departments answered. Follow-up of treatment with amiodarone and euthyroid sick syndrome were considered the only conditions justifying FT3 test ordering. CONCLUSION: Though the opinion of physicians specialized in endocrinology was not uniform regarding recommendations for TT3 or FT3 tests as a first-line measurement, the cost of these tests has been estimated at 650 thousand Francs for a month at the AP-HP. PMID- 10374017 TI - The philosophical basis of the experimental method. PMID- 10374018 TI - Microvascular lung tissue oxygenation--a methodological study in the pig. AB - The objective of the present study was to investigate the possibility of measuring lung tissue oxygen pressure (PtO2) distributions at the microvascular level, and also if a change in the lung tissue oxygenation could be detected during hypoventilation (50% reduction in ventilatory settings). Experiments were carried out on eight mechanically ventilated ketamine-anaesthetised pigs. A thoracotomy was performed through the third right intercostal space. PtO2 measurements were made using a Clark-type multiwire microelectrode placed onto the pleural surface of the middle lobe. PtO2 was measured during normoventilation, hypoventilation (3 minutes) (reduction of respiratory volume/minute and frequency by 50%), and a second period of normoventilation. Baseline PtO2 was 5.8 (range 4.4 to 10.3) kPa and decreased to 2.9 (range 1.6 to 4.2) kPa during hypoventilation, associated with some PtO2 values close to zero. The PtO2 increased to 5.4 (range 3.6 to 8.4) kPa during the second normoventilatory period, some values still close to zero. This study demonstrates that lung tissue PO2 registrations can be made in a suitable animal (pig) model, and that hypoventilation induced an almost reversible decrease in lung tissue oxygen pressure distributions. In addition, no microscopically visible tissue damage was inflicted by the electrode on the underlying lung surface. PMID- 10374019 TI - The results of autogenous tibial periosteal transplants for full thickness cartilage defects in the knee joints of pigs. AB - An experimental study was conducted to demonstrate macroscopic and microscopic healing of full thickness cartilage defects with mature stable hyaline cartilage after autogenous tibial periosteal transplants in the knee joints of pigs. Similar full thickness osteochondral defects were created in the medial femoral condyles of both knees in 10 healthy young adult pigs. Periosteal transplants were performed on the left knees and the right knees used as controls. The pigs were sacrified in two groups at 6 weeks and 3 months. The knees were inspected for healing and stability of the graft. Microscopic sections were taken and evaluated using a histological score developed by O'Driscoll. Macroscopically, almost all defects with periosteal grafts healed with a translucent bluish-white colour indicating articular cartilage formation. There was good restoration of the bony contour and filling of the defects were superior to controls. Histologically, we were able to demonstrate immature hyaline cartilage which matured at 3 months. The newly formed tissue was stable and well-incorporated. It had almost complete bonding to the adjacent articular margin, good reconstitution of the osteochondral junction and a well maintained structural integrity. We concluded that periosteal transplants in the knee of a pig model healed with mature stable hyaline cartilage. PMID- 10374020 TI - A simple method of blood pressure measurement in the pig using a neonatal cuff. AB - The pig is a commonly used large animal model in experimental studies. Few non invasive techniques exist however for the measurement of blood pressure in the porcine model. This study evaluates the novel use of the easily available neonatal blood pressure cuff for measuring blood pressure in the pig. Six Yorkshire pigs were used for the study. Blood pressure measurements obtained by the application of neonatal blood pressure cuff (Hewlett Packard) around the base of the tail were compared with results obtained from intra-arterial measurements in the normotensive range as well as in experimentally created hypertensive (intravenous dopamine) and hypotensive (hypovolaemic shock) ranges. Results of the two techniques are closely correlated (Pearson's coefficient = 0.95, 0.97, 0.90). Systematic bias was however detected at the extremes of hypertensive and hypotensive blood pressure. Analysis of the limits of agreement (method of Bland and Altman) showed that neonatal blood pressure cuff measurements fall within--2 to 2.5 mmHg of the readings obtained from the invasive technique (95% confidence interval). The neonatal blood pressure cuff technique is a good substitute for the standard invasive intra-arterial measurement of blood pressure in the pig model. PMID- 10374021 TI - Use of titanium prosthesis to bridge a vertebral gap in the spine--a preliminary experimental study. AB - Resection of a vertebral body for spine tumour or fracture results in a vertebral gap which has to be bridged by autogenous graft, allograft, bone cement or metal spacer. Recently, there have been several metal spacers in the market. We have designed a titanium vertebral spacer which is extensible by way of a threaded mechanism. Coating with hydroxyapatite enables bone ingrowth onto the surface of the titanium spacer. Biomechanical analysis, using the Instron biaxial electro servohydraulic testing machine, showed that the segment bridging the spacer was rigid and stiffer than the adjacent vertebral body motion segment. Histological study showed that there was bone growth across the vertebral gap indicating fusion had taken place. PMID- 10374022 TI - The viability of liver graft for transplantation after prolonged warm ischaemia. AB - Maximum duration of warm ischaemia within which the liver graft is viable for transplantation remains undefined. Published data on porcine allogeneic liver transplantation (LTx) using non heart beating donors (NHBDs) are conflicting because technical details like the hepatic artery status, systemic heparinisation of donor animals and duration of rewarming were not addressed. We described a novel porcine model which simulate conditions of transplantation from NHBDs. The pigs were divided into three groups of 6 each. Groups I, II and III were subjected to 60, 90 and 120 minutes of warm ischaemia, respectively. Liver viability was assessed using four parameters: serum liver function tests (serum bilirubin and transaminase), dynamic liver function test i.e. the monoethylglycinexylidide (MEGX) formation test, morphological assessment and animal survival. All animals in groups I and II (90 minutes of warm ischaemia or less) survived but 50% of animals in group III died of massive liver failure. Given that rewarming period required in actual allogeneic LTx is about 60 minutes, the safe period for intervention in NHBDs is determined to be about 30 minutes. Allogeneic porcine LTx using NHBDs with 30 minutes of cardiac arrest were performed in 5 animals. All of them survived. PMID- 10374023 TI - Direct mucosal targeting of colonic receptors by prokinetic drugs in an experimental model. AB - Isolated perfused segments of pig ileum and sigmoid colon were used as an extrinsically denervated model of intestinal fluid propulsion to compare the effects of intraluminal (IL) with intraarterial (IA) administration of cisapride and mebeverine. The ileal segments had a spontaneous mean activity of 0.008 (SEM 0.003) ml Krebs propelled aborally min-1, with propulsive waves at a mean frequency of 8.3 (1.6) min-1. The sigmoid colon segments ejected a mean of 0.013 (0.009) ml Krebs min-1, with propulsive waves at 3.9 (0.8) min-1. IL cisapride produced a dose-dependent response in the dose range 1 x 10(-9) M to 3 x 10(-1) & 1-3 x 10(-7) M in the ileum, and 3 x 10(-11) to 3 x 10(-9) M in the colonic segments, IL cisapride was significantly more effective than IA delivery of equivalent doses. IL instilled mebeverine (1 x 10(-6) M) inhibited the carbachol dose response of the ileal and colonic segments more than an equivalent dose of mebeverine infused IA. We conclude that the isolated perfused pig intestine is an effective model for studying the pharmacological effects of drugs and their routes of delivery. Cisapride and mebeverine were more effective per given concentration, when delivered IL than IA in both the ileum and sigmoid colon preparations. The qualitative effects of either IL or IA drug delivery were not affected by extrinsic denervation. PMID- 10374024 TI - The effect of storage at -70 degrees C and -150 degrees C on the torsion properties of the canine femur. AB - This study investigates the effect of storage at -150 degrees C in the vapour phase of liquid nitrogen on the torsion properties of whole intact femurs to be used as allografts. Twenty-five adult dog femurs, stripped of all soft tissue, were used. It has been established that storage at -70 degrees C does not significantly affect the torsion properties of bone. In this study we found no significant difference in structural properties in torsion between the allografts stored at -150 degrees C and -70 degrees C. However, cortical hairline cracks, which were not present before storage, after the bones were stored for 3 months and thawed at 1 degree C/min rate were observed. These cracks fractured longitudinally, and not spirally, when tested to failure and were noted to originate from the vessel foramen. The mean torsion strength of these "damaged" bones was reduced by 48%. We also determined that these hairline cracks occurred during the thawing process at about -42 degrees C. This may suggest an effect of recrystallisation of fluids or water crystals within the "closed" medullary cavity and the foramen, thus increasing the pressure and paying for hairline cracks to propagate from stress risers. No incidence of cortical hairline cracks were observed if the medullary canal was decompressed and the medullary canals washed-out. In tissue banking whole intact bones as allografts, caution should be taken when selecting the freezing and thawing rates for storage as cortical hairline cracks could result and eventually weaken the strength of the allograft. PMID- 10374025 TI - Radionuclide studies of articular cartilage in the early diagnosis of arthritis in the rabbit. AB - The compound Se-75 bis[beta-(N,N,N-trimethylamino-)ethyl]selenide diiodide (Se-75 BISTAES) has been synthesized and its biodistribution in rabbits studied. A high uptake of radioactivity is found in the knee cartilage. Good scans of the knee are obtained by nuclear scintigraphy at 15 minutes after the injection of Se-75 BISTAES. The results of an equilibrium dialysis study show that Se-75 BISTAES binds to chondroitin sulfate and the binding is directly proportional to the chondroitin concentration. It appears that Se-75 BISTAES or its derivative should have potential as an articular imaging agent. PMID- 10374026 TI - Nitric oxide synthase--its distribution and alteration in the intramural ganglia of the urinary bladder in normal and urethra-obstructed guinea pigs. AB - Nitric oxide (NO) has been proposed to function as an inhibitory neurotransmitter in the lower urinary tract. This study investigates the distribution of NO containing neurons and its changes following urethral obstruction in the guinea pig. By using nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry and NO synthase (NOS) immunohistochemistry, the highest frequency of NO-containing neurons was observed in the bladder base. Double labelling studies showed that 70.9% of NADPH-d reactive neurons co-expressed NOS immunoreactivity. Acetylcholinesterase reactivity was present in the majority of the intramural neurons with 54% of them expressed NOS immunoreactivity. NADPH-d reactivity was colocalized with vasoactive intestinal polypeptide, calcitonin gene-related peptide and substance P immunoreactivities in both neurons and fibres. Colocalization study also revealed that NADPH-d reactive neurons formed a distinct cell population from tyrosine hydroxylase positive neurons. At 12 hours after urethral obstruction, NADPH-d reactivity in the intramural ganglion cells was noticeably enhanced and this was sustained till 24 hours whence some intensely stained neurons appeared to undergo degenerative changes. Neuronal degeneration was more drastic at 48 hours so that the number of NADPH-d positive neurons was significantly reduced. The present study suggests that NO is an important neurotransmitter in the urinary bladder and that it may be involved in the relaxation activity in the bladder base during micturition. It is speculated that the increased NADPH-d reactivity in intramural ganglion cells elicited by urethral obstruction may be responsible for the cell death. It is suggested that the resulting cell loss or bladder denervation may account for the urinary dysfunction such as frequency and urgency of micturition in patients with urethral obstruction. PMID- 10374027 TI - The expression of insulin-like growth factor II, hepatitis B virus X antigen and p21 in experimental hepatocarcinogenesis in tree shrews. AB - The purpose of this paper was to study the mechanism of synergistic effect in hepatocarcinogenesis induced by hepatitis B virus (HBV) infection and aflatoxin B1 (AFB1) intake. Immunohistochemical staining was used in formalin-fixed, paraffin-embedded sections of cancer and liver tissues. The incidence of hepatocellular carcinomas (HCCs) was 52.9% in experimental tree shrews that received both HBV and AFB1. It was significantly higher than that of animals exposed to HBV (11.1%, Group B), or (AFB1) (15.8%, group C) alone. HCC was not found in the control animals (group D). The expressions of insulin-like growth factor II (IGF-II) were 82.4%, 22.2%, 26.3% and 0 in groups A, B, C and D, respectively. The significant differences of IGF-II were observed between groups A and B, C and D (P < 0.05). The expressions of p21 were 29.4%, 11.1%, 15.8% and 0 in group A, B, C and D, respectively. The positive rate of hepatitis B x antigen (HbxAg) was significantly higher in the group A than that in the group B (52.9% vs. 11.1%, P < 0.05). The parallel relations between the incidence of HCC and the overexpressions of these genes protein have been found in each group. On the other hand, the expressions of these genes in tumour-bearing tree shrews were significantly higher than that in nontumour-bearing animals. These findings suggest a synergistic effects of HBV and AFB1 in activation of these genes in tree shrews. Overexpressions of these genes may take an important role in the course of hepatocarcinogenesis in tree shrews. PMID- 10374028 TI - Synergistic effect of hepatitis B virus and aflatoxin B1 in hepatocarcinogenesis in tree shrews. AB - An animal experiment with tree shrews was performed to detect the synergistic effects of hepatitis B virus (HBV) infection and dietary aflatoxin B1 (AFB1) in hepatocarcinogenesis. Adult healthy tree shrews (Tupaia belangeri chinensis) were divided into four groups: Group A (HBV + AFB1)--animals were infected with human HBV serum at first, then fed AFB1 diluted with milk, 150 ug/kg.bw/day, 6 days/week for 105 weeks. Group B (HBV)--animals were infected with human HBV as Group A, but no AFB1 treatment. Group C (AFB1)--animals were treated with AFB1 as Group A but no HBV infection. Group D--animals were treated neither with human HBV nor AFB1. During the experiment, blood samples and liver biopsies were taken regularly from all animals in each group. All the animals were sacrificed on the 160th week when the experiment ended. The samples of sera and liver tissues were checked for HBV markers and histological changes. Hepatocellular carcinomas (HCCs) were found only in Group A and Group C, with incidences of 67% and 30% respectively. The average time for HCC occurrence in Group A and Group C was 120.3 +/- 16.6 and 153.3 +/- 5.8 weeks respectively (P < 0.01). Even though no HCC occurred in Group B, 1 animal which died before the end of the experiment showed two large hepatocellular nodules. These results showed that there is synergistic effect between HBV and AFB1 in tree shrews' hepatocarcinogenesis, even though the hepatocarcinogenic effect played by HBV alone is rather weak. PMID- 10374030 TI - The role of mast cell degranulation in ischaemia-reperfusion-induced mucosal injury in the small intestine. AB - The role of the intestinal mast cell system in the pathophysiology of postischaemic mucosal lesions is not understood. The present goals were to investigate the contributions of mast cells and mast cell-derived vasoactive mediators to mucosal injury caused by arterial occlusion. We evaluated the intestinal ischaemia-reperfusion-induced local morphological changes in mast cell depleted anesthetized dogs. Animals subjected to complete segmental intestinal ischaemia and reperfusion served as controls. The selective mucosal-type mast cell degranulator Cremophor-El and the nonselective mast cell depleter Compound 48/80 were used to investigate the involvement of mast cells in reperfusion induced tissue reactions. Ileal biopsies taken at the end of 120 min of ischaemia and after 120 min of reperfusion were evaluated histologically. The number of mast cells was determined and the degree of mucosal damage was evaluated according to the 0 to 5-grade Chiu scale. Mucosal histidine decarboxylase activity was measured in tissue biopsies and the rate of release of histamine was determined from the venous effluent of the segment. In the control group, 120 min reperfusion induced a severe tissue injury. In the Compound 48/80 and Cremophor El-pretreated groups, the reduction in the baseline number of mast cells was 37% and 53%, respectively, and the basal mucosal histidine decarboxylase activity was significantly increased. In these groups, the ischaemia-reperfusion-induced release of histamine was significantly decreased, and the degree of damage of the intestinal mucosa was significantly reduced. Mucosal mast cell degranulation plays an important role in the initiation of tissue injury after intestinal ischaemia-reperfusion. Depletion of mast cells prior to ischaemia decreases the severity of mucosal damage, probably in consequence of the stimulation of mucosal histidine decarboxylase activity. PMID- 10374029 TI - Efficacy of contraction uncoupling by 2,3-butanedione monoxime during initial reperfusion versus cardioplegic arrest for protection of isolated hearts. AB - The efficacy of 2,3-butanedione monoxime (BDM) as additive to St. Thomas Hospital II solution (STH) as compared to initial BDM reperfusion with regard to myocardial ischaemia/reperfusion injury was investigated in isolated guinea pig hearts. Isolated guinea pig hearts were perfused with Krebs-Henseleit buffer in the Langendorff technique at constant pressure of 55 mmHg. After cardioplegic arrest with STH solution, global ischaemia was induced for 50 min and recovery of myocardial function was monitored during 30 min reperfusion. Control hearts (n = 8) received no further treatment. In BDMCP hearts (n = 8), 20 mM BDM were added to STH only. BDMREP hearts (n = 8) were treated with 20 mM BDM during the initial 20 min of reperfusion. BDMCP/REP hearts (n = 8) received BDM during cardioplegic arrest as well as during initial reperfusion. Left ventricular systolic function as assessed by developed pressure (LVDP) was depressed to 47 +/- 3% of pre ischaemic baseline in control. Only initial BDM reperfusion (BDMREP) resulted in improved recovery of LVDP to 66 +/- 5%. Similar data were obtained for dP/dtmax and dP/dtmin. Reperfusion contracture was attenuated in both groups receiving initial BDM reperfusion (BDMREP and BDMCP/REP). BDM in STH did not protect hearts from cellular injury as assessed by release of lactate dehydrogenase (LDH) during reperfusion. In contrast, no increase in LDH release occurred during initial BDM reperfusion in BDMREP and BDMCP/REP hearts, followed by a mild rebound after washout of the drug. Addition of BDM to the cardioplegic STH solution did not protect isolated hearts from cellular injury or depression of post-ischaemic function. In contrast, initial BDM reperfusion alone attenuated reperfusion contracture, prevented LDH release, and improved recovery of systolic and diastolic myocardial function. The combination of BDM treatment during cardioplegic arrest with initial BDM reperfusion provides no additional protection from reperfusion injury. PMID- 10374031 TI - Experimental study of hypovolaemic shock-induced gastric mucosal lesions in the rat. AB - This study was designed to determine whether oxygen-derived free radicals play a role in the pathogenesis of gastric mucosal lesions produced by haemorrhagic shock and reperfusion experimental model in the rat. Ranitidine (H2-receptor blocker) in different doses, allopurinol, an inhibitor of xanthine oxidase and SOD (superoxide dysmutase) pre-treatment were used against haemorrhagic shock and reperfusion induced gastric mucosal lesions. Altogether 67 rats were divided into seven different groups. The area of gastric mucosal lesions was measured, the activity of endogenous peroxidase was examined histochemically and histological grading was made. Evans blue was used to demonstrate the improved permeability of gastric mucosal membranes. Ranitidine, in high dose, allopurinol and superoxide dysmutase significantly protected against haemorrhagic shock-induced gastric mucosal lesions, against improved membrane permeability and peroxidation. PMID- 10374032 TI - Establishment of animal models using experimental rats for allogeneic tissue transplantation and quantitative flow cytometric detection of immunochimera. AB - Establishment of animal models used for allogeneic tissue transplantation, MHC phenotyping as well as quantitative detection of immunochimera were carried out in this study. Both Buffalo (BUF) and Lewis (LEW) rats were chosen as the target animals. The rats were treated with single dose cyclophosphamide (Cy) of 25 to 100 mg/kg to mimic the standard conditioning therapy. Total white blood cells (WBC) were monitored daily for up to 8 days: WBC reached the nadir by day 4 and started to recover by day 5 with an obvious rebounce at day 7 of Cy treatment. Flow cytometric techniques were used to determine the haplotypes of the major histocompatibility complex (MHC) as well as quantitative detection of immunochimerism in unfractionated rat WBC. Monoclonal antibodies against the rat class-I MHC antigens RT1Aab and RT1Au were used to label the class-I MHC antigens on total rat WBC. The results showed that the BUF rats were positive for both RT1Aab and RT1Au antigens, whilst the LEW rats were negative for both. Immunochimera was mimicked in vitro by serial dilution, ranging from 1/1 to 1/10(5) of (BUF/LEW) WBC. A sensitivity of 1/10(4) (BUF/LEW WBC) was achieved. The results showed that there were at least 2 major MHC mismatched loci between BUF and LEW rats and flow cytometry provided a sensitive method for the detection of immunochimera in unfractionated rat WBC. We concluded from this study that both strains of rats could be used as models for allogeneic tissue transplantation across at least two major MHC-mismatches. The sensitivity of flow cytometric method was satisfactory for the detection of immunochimera. PMID- 10374033 TI - An animal model for the study of hepatic stellate cell and hepatocellular carcinoma interaction. AB - The recognition of hepatic stellate cells (HSC) around and within hepatocellular carcinoma (HCC) in human livers has generated interest in the interactions between HSC and HCC. We explored the possibility of creating an animal model to allow in vivo investigations of this interaction. Eighteen adult Buffalo rats were inoculated with 1 x 10(6) cells obtained from cultures of Morris 7777 hepatoma cell line (ATCC). The rats were sacrificed at 2-, 3-, and 4-week intervals. Identification of activated HSC was with immunohistochemistry for alpha-smooth muscle actin (alpha-SMA). There was 100% survival of all animals until sacrifice. Tumour formation occurred in 94.4% of rats, and was of a good size by two weeks. Expression of alpha-SMA was observed around and within all HCC, but absent from normal tissue, and this showed colocalisation with collagen deposition. These findings are consistent with those previously reported in resected HCC in humans. The high survival, good tumour yield, consistent generation of activated HSC around the tumours, and similarities in histological appearance to the human HSC-HCC distribution pattern, make this a reliable animal model for in vivo studies on HSC-HCC interaction. PMID- 10374034 TI - Regulation of insulin secretion by nerves and neuropeptides. AB - The pancreatic islets are innervated by sympathetic, parasympathetic and sensory nerves and within the terminals of these nerves several different neurotransmitters are stored, both the classical neurotransmitters, acetylcholine and noradrenaline, and several neuropeptides. The neurotransmitters affect insulin secretion both in a stimulatory and in an inhibitory direction depending on the nervous system activated, the neurotransmitter released and the status of the B cell. This neural regulation of insulin secretion is of importance during various forms of stress as well as during the cephalic phase of food intake. PMID- 10374035 TI - Development of the human intrahepatic biliary system. AB - In the development of the human biliary system, the extrahepatic bile ducts (EHBD) develop from the embryonic hepatic diverticulum, while the intrahepatic bile ducts (IHBD) originate within the liver from the ductal plate. The ductal plate is a flat muralium of primitive biliary epithelium that develops in the mesenchyme along the branches of the portal vein, by a process which requires a delicate balance between cell proliferation and death. The ductal plate is thus remodelled into the adult system of tubular anastomosing bile ducts and this process is called ductal plate remodelling. Computerised three-dimensional reconstruction of the developing ductal plate has shown that the ductal plate remodelling process starts at the porta hepatis around 11 weeks of gestation and progresses towards the periphery of the liver. The extrahepatic biliary system is in direct luminal continuity with the developing intrahepatic biliary system throughout gestation and does not have a "solid stage" as suggested previously. The ductal plate remodelling is controlled by many biochemical and molecular factors, some of which have been identified and studied. It has been suggested that abnormalities in the development of the IHBD could lead to a spectrum of diseases called ductal plate malformation. Biliary atresia is one of the conditions in this spectrum. Currently, we are studying the IHBD in biliary atresia in comparison to the normal developing IHBD, the results of which are presented in this review. Both morphologically and biochemically the IHBD in biliary atresia resembles the primitive foetal ductal plate suggesting a disruption in ductal plate remodelling. PMID- 10374036 TI - Experimental models of hepatic fibrosis in the rat. AB - The rat is a frequently used experimental model in studies involving human disease. We review several methods of inducing hepatic fibrosis and cirrhosis in rats. These include induction by hepatotoxins and hepatocarcinogens such as carbon tetrachloride, dimethylnitrosamine, thioacetamide and furan; the hepatoxin cum-nutrient, alcohol; a high fat-low choline-low protein diet; immunologic agents such as heterologous serum or bacterial cell wall products; and obstructive jaundice and biliary cirrhosis by common bile duct ligation. PMID- 10374037 TI - Murine metastatic tumour models for cancer gene therapy research. AB - The appropriate use of animal tumour models has a pivotal role in the evaluation of any new anti-cancer therapy. Indeed, animal models of human diseases have been emphasised in the early development and evaluation of gene therapy. Given that most cancer treatment failures and cancer-related mortality are the direct results of metastatic cancers, the increasing use of clinically-relevant metastatic tumour models in the study of cancer therapeutics is both logical and necessary. Murine metastatic tumour models may be established either "experimentally" or "spontaneously". The yield and reproducibility of spontaneous metastasis models can be enhanced through manoeuvres such as using highly metastatic sublines for primary tumour implantation and orthotropic transplantation. The use of immunodeficient rodents, although popular, suffers from the absence of T-cell responses in the host which may impact on therapeutic efficacy. While many gene therapy strategies today are capable of regressing primary tumours in experimental animals, only a limited number of approaches (viz. genetic immunotherapy and gene-mediated anti-angiogenesis) are designed to address the challenges posed by metastatic tumours. In extrapolating the results of gene therapy in animal models to humans, it is important to appreciate the heterogeneity of the latter populations, and anticipate greater variability in the treatment outcome. PMID- 10374038 TI - Immunological consequences of trauma and shock. AB - The immune system is a powerful, complex entity composed of numerous cell types and regulated by autocrine, paracrine, and hormonal mechanisms. Trauma and haemorrhagic shock induce numerous changes within this system which are ultimately deleterious and contribute to the high incidence of organ dysfunction and infectious complications seen following injury. Regional hypoxia and depletion of intracellular energy stores occur in response to diminished microcirculatory blood flow, and these changes alter cellular signalling and result in the release of pro-inflammatory cytokines and prostanoids which mediate further suppression of immune function. Neutrophil priming serves to induce tissue damage in critical organ systems such as the lungs, heart, liver, and gut, further insulting the injured organism. Depression of antigen presentation and cytokine elaboration by macrophages and other antigen presenting cells effectively prevents a normal response from the acquired immune system, and lymphocyte-monocyte interactions are squelched. The resulting depression in cell mediated and humoral immunity renders the organism susceptible to microbial infection and contributes to the morbidity and mortality associated with nosocomial infections. Hormonal modulation of the immune response is highly evident following trauma and haemorrhage, and the preponderance of male morbidity associated with sepsis can be explained by the depression in immune function seen in males, but not females in the pro-oestrous state. Despite the multitude of changes induced by trauma and haemorrhage, experimental studies have revealed several promising pharmacologic interventions which may serve to blunt the effect of injury on the immune system, and render the host competent to withstand the bacterial and viral challenges responsible for so much of the late mortality following severe injury. PMID- 10374039 TI - Experimental models of pancreatitis. AB - Experimental animal models are helpful tools that have been employed to study pancreatitis for more than a century. Although not closely related to all aspects of the human disease, they have contributed greatly to our current understanding of the pathophysiology and cell biology of this disease. They have also become a standard means of testing innovative treatments against pancreatitis. This article reviews the experimental models of acute pancreatitis in common use, their severity, the criteria for the selection of an appropriate model, standards in monitoring and relevance to clinical disease. Despite their undoubted value in elucidating the mechanisms involved in the early cellular events and pathophysiology of acute pancreatitis, these models have not lent themselves to the development of effective new therapies. Models of chronic pancreatitis are also reviewed. The development of a reproducible model relevant to human chronic pancreatitis remains challenging. Experimental models of chronic pancreatitis have not yet added greatly to the understanding of the pathogenesis of this disease in man. PMID- 10374040 TI - Gut barrier dysfunction in experimental acute pancreatitis. AB - Bacterial infections, frequently caused by bacteria of enteric origin, are often seen during the progression of severe acute pancreatitis, concomitant with the potential development of multiple organ dysfunction. In the present paper, the complex mechanisms of gut barrier dysfunction and gut origin sepsis in acute pancreatitis are discussed. The individual parts of the gut barrier, e.g. the immunological, bacteriological and morphological (mucus, epithelium, endothelium and interstitium) components, seem to interact and depend on each other. Pancreatitis-induced hypovolaemia due to endothelial barrier leakage and gut arteriovenous shunting causes intestinal ischaemia and reperfusion injury with concomitant gut barrier dysfunction. Gut endothelial barrier dysfunction probably plays a central role. Potential molecular mechanisms could, among others, be associated with alterations in intracellular signal transduction, intercellular signalling and expression of adhesion molecules on endothelial cells. The mechanisms underlying gut barrier dysfunction in acute pancreatitis are thus complex and still not fully elucidated. Knowledge about the regulating events will probably allow future pharmacological therapy for prevention and treatment of the severe complications of acute pancreatitis, including gut barrier dysfunction. PMID- 10374041 TI - Effect of antiangiogenic agents on experimental animal models of hepatocellular carcinoma. AB - A new therapeutic strategy for treating metastasis in hepatocellular carcinoma (HCC) has entailed the use of antiangiogenic agents such as suramin, BB-94 (Batimastat), TNP-470, and carboxyamido-triazole (CAI, a synthetic inhibitor of non-excitable calcium channels that reversibly inhibits angiogenesis). These agents have been used to treat metastatic model of HCC in nude mouse (LCI-D20 mouse model). The results of these studies are summarized in this paper with emphasis on the inhibitory effects of the drugs on tumour growth, angiogenesis, invasion and metastasis in LCI-D20 mouse models. The results suggest that all of the agents used can significantly inhibit tumour growth, angiogenesis, invasion and metastasis of human HCC in nude mouse models, and may be candidates for the control of recurrence and metastasis after HCC resection. PMID- 10374042 TI - Tissue microangiography using a simplified barium sulphate cadaver injection technique. AB - This study investigated the application of soft tissue X-ray imaging as a method of studying vascular anatomy in injected cadaveric specimens. The arterial system of the anatomical region of interest was perfused with a barium sulphate-gelatin suspension. After curing, the tissues were removed by meticulous dissection and examined using the soft tissue X-ray imaging system. This technique of microangiography was able to demonstrate the entire arterial system of the perfused specimen. This simplified technique shows great potential in the study of vascular anatomy of flaps. PMID- 10374043 TI - A practical technique of colour image analysis: applications in experimental research. AB - Commercially available colour image analysers are relatively expensive. We describe a cheaper alternative developed by the Department of Experimental Surgery, Singapore General Hospital using an assembly of optical and computer equipment commonly available in the research laboratory. This manual colour imaging system is comparable to the commercial model in terms of functional capabilities and accuracy, except that it takes a longer time to process and analyse images and is unable to measure colour density. However, it is capable of not only analysing microscopic images of stained histological tissue sections but also X-ray images and images of large pathological specimens. In the case of commercial models, different systems have to be used to analyse images from different types of specimens. This system was developed in 1987 and has since been used successfully in a number of experimental studies. It has been applied to the measurement of parameters defining eye anatomical configuration, delineating the extent of tissue necrosis and fibrosis after therapeutic treatment and surgery, the development of new bone formation in fracture healing and to quantitative studies in liver regeneration. Due to its accuracy, low cost and versatility, this system should be within the means of even the most modest research laboratory. PMID- 10374044 TI - What is atopy? Condition, disease or a syndrome? PMID- 10374045 TI - Epidemiology of atopic dermatitis: recent advances and future predictions. PMID- 10374046 TI - Association of birch pollen-related food-responsive atopic dermatitis with birch pollen allergen-specific T-cell reactions. PMID- 10374047 TI - What do we know about the etiopathology of the intrinsic type of atopic dermatitis? PMID- 10374048 TI - Role of T cells and cytokines in the intrinsic form of atopic dermatitis. PMID- 10374049 TI - Autoallergy: a pathogenetic factor in atopic dermatitis? PMID- 10374050 TI - Neuropeptides in atopic dermatitis. PMID- 10374051 TI - Advances in the treatment of atopic dermatitis with special regard to children. PMID- 10374053 TI - Food allergy: a diagnostic challenge. PMID- 10374052 TI - Role of food allergy and food intolerance in recurrent urticaria. PMID- 10374054 TI - Genetically modified food: a danger or a benefit for atopics? PMID- 10374055 TI - Atopic and vernal keratoconjunctivitis: a model for studying atopic disease. PMID- 10374056 TI - New insights in the pathogenesis of allergic rhinitis. PMID- 10374057 TI - Advances in the pharmacological treatment of allergic rhinitis. PMID- 10374058 TI - Immunotherapy in allergic rhinitis: a prevention for asthma? PMID- 10374059 TI - New insights into the pathogenesis of asthma. PMID- 10374060 TI - Atopic and non-atopic asthma. PMID- 10374061 TI - Advances in pharmacotherapy of asthma. PMID- 10374062 TI - The environment and the skin. PMID- 10374063 TI - The atopic child and the environment. PMID- 10374065 TI - Perspectives of atopic eczema in the third millennium. PMID- 10374064 TI - Primary and secondary prevention of atopic diseases in children. PMID- 10374066 TI - Catecholamine release from isolated guinea pig lungs during sympathetic stimulation with varied ventilation and perfusion. AB - This study was performed to elucidate catecholamine release in the pulmonary circulation of isolated lungs due to the sympathetic nerve stimulation and to assess the experimental conditions which can modify the release, i.e., stimulus intensity, ventilation state of the lung and flow rate of perfusion. In artificially ventilated lungs, electrical stimulation of stellate ganglions evoked large noradrenaline efflux from the lung, but adrenaline efflux was below the detection limit, and dopamine was not detected in any case. In the unventilated preparations, the lung parenchyma were not bleached and the arterial pressure was significantly higher than in ventilated preparations. Noradrenaline efflux from the unventilated group was significantly lower than that from the ventilated preparations. The effect of the perfusion flow rate was investigated under pressure-operated ventilation. The pulmonary arterial pressure (Pa) was not varied at 5-10 ml min-1, but it was increased significantly at 20 ml min-1. Noradrenaline efflux was also increased significantly at 20 ml min-1. These results indicate that noradrenaline was the catecholamine exclusively released from pulmonary vasculature due to the sympathetic nerve stimulation, and that both ventilation and the perfusion flow rate could affect the release. The concomitant increase in arterial pressure indicates that noradrenaline efflux would be affected by the alteration in resistive small arteries. Circulatory change in these arteries is supposed to be one of the factors that modify noradrenaline release from the lungs. The analysis of noradrenaline should be a useful method to evaluate the sympathetic effect on the pulmonary vasculature. PMID- 10374067 TI - Tawny: a novel light coat color mutation found in a wild population of Mus musculus molossinus, a new allele at the melanocortin 1 receptor (Mc1r) locus. AB - We found a new coat color mutant in a population of Japanese wild mice (Mus musculus molossinus) and called the trait tawny. The tawny mutant is characterized by a light yellowish brown coat color. The tawny hair has a so called agouti pattern, but the yellow band is greatly lengthened. There are no differences between the tawny and wildtype hairs in size and the number of melanosomes. Genetic analyses revealed that the tawny trait is an autosomal recessive and its gene is located in the distal region on Chromosome 8 between the microsatellite markers D8Mit87 and D8Mit122. An allelism test indicated the tawny mutant gene to be a new allele at the Mc1r locus and dominant to the recessive yellow (Mc1re). The proposed gene symbol for the tawny is Mc1rtaw. PMID- 10374069 TI - Peripheral mononeuropathy induced by loose ligation of the sciatic nerve in the rat: behavioral, electrophysiological and histopathologic studies. AB - The relationship between clinical parameters and pathological changes was investigated in an animal model of mononeuropathy, by behavioral, electrophysiological and histopathological methods. Mononeuropathy was induced in rats by loosely tying ligatures around the sciatic nerve. Eighty-four rats were used, and these were divided into fourteen groups to determine chronological changes in the withdrawal reflex latency, nerve conduction velocity and ultrastructure of the nerve from 1 to 84 days after nerve ligation surgery. Pathological changes around the ligated nerves were divisible in three phases: the first week was an inflammatory phase, when axonal degeneration, phagocyte infiltration and interstitial edematous changes were observed. The second and third weeks were a nerve-sprouting phase, when numerous axonal sprouts and remyelination were seen. The fourth to twelfth weeks were a recovery phase in which maturing myelination and interstitial fibrosis were characteristic. In the inflammatory phase, withdrawal reflex latencies were shortened, and sensory nerve conduction velocities (SCV) and motor nerve conduction velocities (MCV) gradually decreased. In the nerve-sprouting phase, the latency values remained low, and SCV and MCV were minimal. The parameters examined gradually returned to control levels during the recovery phase. In conclusion, these findings increase the knowledge of disease progression in mononeuropathy with hyperalgesia in human and animal models. PMID- 10374068 TI - Production of ex-germfree rabbits for establishment of specific pathogen-free (SPF) colonies. AB - Nine groups of ex-germfree (GF) rabbits were produced by inoculation of hysterectomy-derived GF rabbits with various combinations of cecal bacteria isolated from conventional (CV) rabbits in order to establish a barrier-sustained colony. Six strains of Bacteroides and two strains of Streptococcus isolated from CV rabbits (2 to 3 weeks old) were used for pretreatment. The mortality of ex-GF rabbits inoculated with the anaerobic growth (CF) on EG or SM10 plates inoculated with a 10(-5) dilution of cecal contents was 71.4 to 94.4% when given without pretreatment. All ex-GF rabbits pretreated with Bacteroides alone survived, but the mortality of ex-GF rabbits inoculated with Bacteroides plus Streptococcus strains as pretreatment was 20 and 45.4%. The mortality of ex-GF rabbits inoculated with only Bacteroides was 43%. All ex-GF rabbits inoculated with Bacteroides plus anaerobic growth (CF), cecal suspension of ex-GF mice which had been inoculated with cecal suspensions of CV rabbits (MF) or chloroform-treated cecal suspension (CHF) survived, but CHF inoculated ex-GF rabbits were in an unhealthy condition with slight diarrhoea. These data indicate that inoculation with Bacteroides strains as pretreatment plus CF or MF was required to convert GF rabbits to the normal state. PMID- 10374070 TI - Blood biochemical characteristics, cecal microbiota and short-chain fatty acid composition in fistula implanted rats. AB - We raised an experimental rat implanted with a cecal fistula and investigated various characteristics of fistula-implanted rats. Male F344/N Sic rats at 14 weeks of age were divided into three groups, the fistula group (n = 5) which consisted of fistula-implanted rats, the sham group (n = 7) which consisted of sham-operated rats, and the control group (n = 7) which were not subjected to any surgical procedure. Four weeks after the fistula implantation surgery, we compared the blood biochemical indices, the microflora composition and the short chain fatty acids (SCFA) concentration in cecal contents of fistula-implanted rats with those of sham-operated and control rats. The blood albumin concentration of the fistula group was significantly lower than that of the sham group and the control group, and the hematocrit value of the fistula group was significantly lower than that of the control group, but there were no significant differences in the SCFA concentration and the microflora composition among these three groups. In conclusion, it was considered that the fistula-implanted rats are useful for taking cecal contents and determining the microflora composition and the metabolites concentration at any time, without disturbing the physiological functions of the intestinal tract. PMID- 10374072 TI - Leukocyte differential analysis in multiple laboratory species by a laser multi angle polarized light scattering separation method. AB - Leukocyte differential analysis was performed in various species, particularly laboratory animals, by the laser multi-angle polarized light scattering separation method. Venous blood specimens were drawn from the following subjects: healthy adult men and women ("humans"); cynomolgus monkeys ("monkeys"); common marmosets ("marmosets"); beagle dogs ("dogs"); miniature potbelly pigs ("swine"); Japanese white rabbits ("rabbits"); Hartley guinea pigs ("guinea pigs"); and Sprague-Dawley rats ("rats"). 90 degrees/10 degrees scatter plot: Basophils and mononuclear-polymorphonuclear cells were separated in all subjects, but individual 10 degrees and 90 degrees scatter plots overlapped in dogs and guinea pigs, respectively. 90 degrees depolarized /90 degrees scatter plot: Neutrophils and eosinophils were clearly separated in human, monkey, guinea pig, swine and rat subjects. The eosinophil cluster was not clearly plotted in marmoset, dog, or rabbit. 0 degree/10 degrees scatter plot: Regarding this plot for monocytes and lymphocytes, cells were plotted in the following order in all subjects: lymphocytes < basophils < or = monocytes in the 0 degree (size) scatter; and lymphocytes [symbol: see text] monocytes < or = basophils in the 10 degrees (complexity) scatter. Compared to other species, the rat scatter showed a tendency to overlapping plots in both the 0 degree and 10 degrees scatters in the monocyte and lymphocyte clusters. In both dog and guinea pig, the monocyte and neutrophil plots overlapped in the 0 degree and 10 degrees scatters. Basobox: In the human and rabbit subjects, the basophil cluster was plotted within the established basobox, but no clear cell cluster was plotted in the other subjects. As a result of comparing the percentage values for leukocytes in various species obtained by using the CD3500 apparatus versus the corresponding values obtained manually, good correspondence was found in the monkey, and eosinophils in the marmoset were lower with CD3500 than manually. In the rabbit, the mean measured value for basophils matched in the manual and CD3500 findings. In the guinea pig, the CD3500 values were lower than the manual values for lymphocytes, but higher for monocytes and neutrophils. The above findings suggest that the laser multi angle polarized light scattering separation method is indeed capable of analyzing leukocytes from various species based on cell size and cell complexity, i.e., the presence or absence of nuclei, granules and cell enclosures. PMID- 10374071 TI - Establishment of specific pathogen-free rabbit colonies with limited-flora rabbits associated with conventional rabbit flora, and monitoring of their cecal flora. AB - In the present study we attempted to establish specific-pathogen-free (SPF) rabbit breeding colonies with two groups of limited-flora (LF) rabbits, both ex germfree rabbits, and their offspring. Two groups of LF rabbits associated with cecal flora of conventional (CV) rabbits produced in a previous study [Exp. Animals, submitted], were transferred to individual barrier rooms and some of the LF rabbits were accommodated in isolators to maintain the basic flora for SPF rabbits. The composition of the cecal flora of LF rabbits was stable for a long period; bacteroides remained predominant and clostridia dominant. From the SPF rabbits, different types of bacteria, e.g., enterobacteriaceae and streptococci, which could not be isolated in the isolator were detected at a low population level at an early stage in the establishment of the SPF colonies, but the basic composition of the cecal flora was mainly bacteroidaceae and clostridia and did not change over a long period, and the floral composition became similar to that of CV rabbits. The fertility and weaning rates of the SPF rabbits were satisfactory for a SPF rabbit colony. In addition, these SPF colonies were free of more than one year rabbit-specific pathogens. PMID- 10374073 TI - Diurnal changes in intraepithelial lymphocytes (IELs) in the small intestine of mice. AB - Diurnal changes in small intestinal intraepithelial lymphocytes (IELs) were examined in 8- to 10-week-old BALB/cA male mice. The ratio of T cell subsets expressing CD8 alpha alpha homodimer/CD8 alpha beta heterodimer was found to be higher in the dark period than that in the light period. Increased expression of Thy-1.2 on gamma delta T cells was also observed in the light period. No significant changes were found in other subsets. This is the first report to document diurnal changes in the small intestinal IELs in mice. PMID- 10374074 TI - Leishmania amazonensis infection in nude mice. AB - Leishmania amazonensis is an intracellular protozoan parasite of macrophages. Cutaneous leishmaniasis in an immunocompetent host begins as papules or nodules followed by ulceration at the site of promastigote inoculation. In this study, the pathological changes of cutaneous leishmaniasis lesions in T cell deficient nude mice were examined. When infected with L. amazonensis promastigotes, nude mice developed non-ulcerative cutaneous nodules. By histological examination of cutaneous lesions, massive accumulation of vacuolated histiocytes containing amastigotes was observed in all the nude mice. Although infiltration of mononuclear and polymorphonuclear cells was seen in the lesions of immunocompetent mice, few such cells were observed in the lesions of nude mice. These results indicate the importance of T cells on the ulcer formation in cutaneous leishmaniasis. PMID- 10374075 TI - Congenital osteoclast deficiency in osteopetrotic (op/op) mice is improved by ovariectomy and orchiectomy. AB - We examined the changes in the appearance of osteoclasts in the femora of ovariectomized (OVX) or orchiectomized (ORX) op/op mice. Osteoclasts on the trabecular bone surface of the OVX or ORX op/op mice significantly increased in number seven or eight times in comparison with sham-operated op/op mice. Furthermore, TRAP-positive cells increased about four times in 100-week-old females and males, compared with sham-operated groups. These results have indicated that a sex hormone reduction due to OVX or ORX induces prominent recruitment of osteoclasts in op/op mice. PMID- 10374076 TI - Induction of pseudopregnancy in the mongolian gerbil (Meriones unguiculatus) by vaginal stimulation. AB - In rats, pseudopregnancy has been induced by mating with vasectomized males, by mechanical stimulation of the uterine cervix with a glass rod or vibrator, and by stimulation of the vagina with a tampon. On the other hand, no practical data are available in reports on the induction of pseudopregnancy in Mongolian gerbils. Pseudopregnancy of gerbils has been induced by mating with vasectomized males. But this method was uncertain because the incidence of pseudopregnancy was lower than that obtained in rats by other means. In the present study, two experiments were undertaken as follows. 1) Copulatory behavior of gerbils was observed for one hour to determine the most effective stimulation interval. 2) From the results of Experiment 1, female gerbils in estrus were mechanically stimulated to test the effectiveness of inducing pseudopregnancy by vaginal stimulation at various time intervals. The results of these experiments indicated that, although the frequency of copulatory behavior varied among individuals, on average the most effective method for inducing pseudopregnancy was stimulation of 5 min duration and at 20 or 30 min intervals. Because the incidence of pseudopregnancy induced by such mechanical stimulation (83.3%) was higher than that induced by mating with vasectomized males (30.0%), this method might be useful in inducing pseudopregnancy in Mongolian gerbils. PMID- 10374077 TI - Evaluation of ready-to-use liquid reagents for clinical chemistry in laboratory animals. AB - We evaluated the ready-to-use liquid reagents for clinical chemistry (6 tests), to assess their suitability for use in the toxicology laboratory setting. Hitachi 736 automated analyzer was used for the analyses. The evaluation included the following studies: Precision, Linearity, Effects of interference substances such as hemolytic hemoglobin, bilirubin, turbidity to the analytical values and correlation to the solid reagents, which are prepared each time they are needed. The precision and linearity data were within the reagents' specifications. Results of comparison of the liquid reagents and the solid reagents in analyzing plasma samples of rats, dogs and monkeys were generally good except for a bias in results for GOT and GPT, regardless of the animal species tested. It is concluded that these types of liquid reagents can be used in clinical pathology examinations in animal studies. PMID- 10374078 TI - Compendium of food additive specifications. Addendum 6. Joint FAO/WHO Expert Committee on Food Additives. 51st session. Geneva, Switzerland, 9-18 June 1998. PMID- 10374079 TI - Manuals of food quality control. 17. Unacceptable visible can defects--a pictorial manual. Codex Alimentarius Committee on Food Hygiene. PMID- 10374080 TI - Screening for cystic fibrosis. PMID- 10374081 TI - Assessing the quality of reports of randomised trials: implications for the conduct of meta-analyses. PMID- 10374082 TI - Is the success of human IVF more a matter of genetics and evolution than growing blastocysts? PMID- 10374083 TI - The use of blastocyst culture to avoid inheritance of an abnormal paternal genome after ICSI. PMID- 10374084 TI - Blastocyst culture and transfer. PMID- 10374085 TI - The combined oral contraceptive pill--are poor communication systems responsible for loss of confidence in this contraceptive method? PMID- 10374086 TI - Gonadal tissue cryopreservation in boys with paediatric cancers. AB - In considering gonadal tissue cryopreservation in children about to undergo chemotherapy or radiotherapy for cancer, numerous issues need to be confronted. This relates firstly to a dearth of information available on the subject of children's gonads coupled to ill-defined fertility preservation procedures, technologies and perhaps less well-publicized regulatory, ethical and legal controls. There may be benefits in considering gonadal tissue preservation for children prior to chemotherapy or radiotherapy, in the hope that future technologies can utilize their immature gametes. Whether or not freezing of gonadal tissue is encouraged prior to cancer therapy, there is a growing demand from parents and clinicians for more information to be made available. This paper reviews our current knowledge of children's gonads in terms of physiology and of fertility experience gained after childhood cancer treatment. It further examines the various issues and concerns regarding children's gonadal tissue storage, its potential use, the present law and the demands and pressures under which clinicians find themselves with patients and parents. This information is important for both the counselling process and decision-making when presented with potential fertility issues in paediatric oncology patients. PMID- 10374087 TI - Endocrine abnormalities during the follicular phase in women with recurrent spontaneous abortion. AB - The frequency of endocrine abnormalities during the follicular phase in non pregnant women with a history of recurrent abortion was investigated in a case control study. A total of 42 consecutive women with recurrent spontaneous abortion (three or more consecutive abortions, mean +/- SD: 3.9 +/- 1.1 range 3 8) with no parental chromosome rearrangement or uterine abnormality were studied during the early follicular phase under standardized conditions. Serum concentrations of follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin, androstenedione, testosterone, dehydroepiandro-stenedione, 17-OH progesterone, oestradiol, progesterone and thyroid stimulating hormone (TSH) were measured by commercially available radioimmunoassays. Controls were 42 nulligravid females with tubal or male factor infertility without miscarriage. Mean (SD) concentrations of prolactin and androstenedione were 14.2 +/- 6.7 ng/ml versus 10.5 +/- 3.5 ng/ml (95% CI 0.8-6.1) and 2.3 +/- 0.9 ng/ml versus 1.7 +/- 0.6 ng/ml (95% CI 0.2-0.9) in the study and control groups respectively. The other endocrine parameters were comparable in both groups. Obesity [BMI weight (kg)/height (m2) > or = 25] was more prevalent (23 versus 5 women, P = 0.0001) in the study than the control group. Recurrent spontaneous abortion is associated with abnormalities in prolactin and androgen secretion during the follicular phase, suggesting an endocrine aetiology in this disorder. Reduction of body weight and correction of hyperprolactinaemia and of hyperandrogenism may reduce the rate of miscarriage in a subsequent pregnancy in these women. PMID- 10374088 TI - FSH inhibits the augmentation by oestradiol of the pituitary responsiveness to GnRH in the female rat. AB - The effect of follicle stimulating hormone (FSH) treatment on the pituitary response to gonadotrophin-releasing hormone (GnRH) was studied in rats in various reproductive conditions. A 3-day treatment of cycling rats with FSH (Metrodin; 10 IU/injection) lowered the spontaneous pre-ovulatory. LH-surge and suppressed the pituitary luteinizing hormone (LH) response to GnRH. FSH also suppressed the LH response of pseudopregnant (PSP) rats on day 8 of pseudopregnancy, but not that of day-8 PSP rats which had been ovariectomized on day 4 (OVX-PSP rats). GnRH induced self priming in cycling, PSP and OVX-PSP rats. Oestradiol strongly augmented the pituitary LH-response to GnRH injection in PSP and OVX-PSP rats, but not in cycling rats; probably because in these latter animals the LH response to GnRH was already augmented by endogenous oestradiol. FSH suppressed the LH response to GnRH in oestradiol-treated PSP and cycling rats; in these latter rats the suppression of the LH response was as strong as that in cycling rats not treated with oestradiol. FSH did not suppress the LH response of oestradiol treated OVX-PSP rats. The effect of FSH was not associated with changes in plasma oestradiol and progesterone concentrations. Analysis of the data revealed that FSH specifically suppressed the augmentative effect of oestradiol, but did not affect the GnRH-self priming effect. It is concluded that under the influence of FSH, the ovaries produce a factor which suppresses the augmentative effect of oestradiol on the GnRH-induced LH response of the pituitary gland. It is suggested that this effect of FSH underlies the suppression of the spontaneous LH surges of FSH-treated cycling rats. As the present putative 'oestrogen antagonizing factor' did not suppress the GnRH-self priming effect, it is suggested that this factor is not identical to gonadotrophin surge inhibiting factor. PMID- 10374089 TI - Nitric oxide reverses prostaglandin-induced inhibition in ovarian progesterone secretion in rats. AB - The present studies were undertaken to determine whether nitric oxide (NO) is involved in the regulation of ovarian progesterone and oestradiol secretion in rats. Immature female Sprague-Dawley rats at 27 days of age were injected s.c. with 4 IU pregnant mare's serum gonadotrophin (PMSG) and were killed 72 h after the injection. The ovaries were collected, weighed and cultured in Dulbecco's modified Eagle's medium containing saline, NO donor, NO synthesis inhibitor or prostaglandin F2 alpha (PGF2 alpha). After 24 h culture, the medium concentrations of progesterone and oestradiol were measured by radioimmunoassay. Results showed that: (i) diethylenetriamine (DETA)/NO (1 x 10(-6), 1 x 10(-5), 1 x 10(-4) M), an NO donor, caused a dose-dependent increase in progesterone synthesis (355 +/- 43, 443 +/- 46, 647 +/- 55 ng/g ovary respectively, P < 0.01) with a concomitant decrease in ovarian oestradiol secretion (408.1 +/- 50.7, 272.9 +/- 28.2, 132.6 +/- 34.6 pg/g ovary respectively, P < 0.01); (ii) neither progesterone nor oestradiol concentrations in the culture medium were altered by DETA without NO; (iii) NG-nitro-l-arginine methyl ester (1 x 10(-4) M), an inhibitor of NO synthesis, did not significantly affect progesterone and oestradiol secretion by rat ovaries; (iv) PGF2 alpha(1 x 10(-6) M) caused a fall in progesterone and oestradiol synthesis; (v) co-incubation with DETA/NO, significantly reversed the PGF2 alpha-induced decrease in progesterone concentrations from 184 +/- 29 to 388 +/- 60 ng/g (P < 0.01), but not that of oestradiol. It can be concluded that NO up-regulates progesterone secretion and down-regulates oestradiol secretion in rat ovaries, and NO can reverse PGF2 alpha induced inhibition in ovarian progesterone secretion. PMID- 10374090 TI - Genetic evaluation of infertile men. AB - Recently, microdeletions in the azoospermic factor region of the Y chromosome, in addition to chromosomal anomalies, have been detected in men with azoospermia or severe oligozoospermia. In this study we evaluated the molecular and cytogenetic defects of infertile men. The frequency of Y microdeletions among 105 azoospermic, 28 oligozoospermic and 32 fertile men was tested on lymphocyte DNA using a series of 20 sequence-tagged sites. In addition, microdeletions were evaluated on testicular-derived DNA among 26 azoospermic patients who underwent testicular biopsy and in whom no sperm cells could be identified. Karyotype analysis was performed on 72 of the infertile patients. Deletions were detected in 6.7% azoospermic and 3.6% oligozoospermic men. No deletions were identified among the fertile men. Identical results were obtained with DNA derived either from lymphocytes or testicular tissue. The frequency of chromosomal aberrations in the 72 infertile patients tested (62 azoospermic, 10 oligozoospermic) was 16.6%, with a high percentage of gonosome anomalies. Additional andrological parameters (hormone values, cryptorchidism) failed to identify men at risk for having microdeletions before the test. Our findings support the recommendation to perform genetic defect screening among infertile men before their enrollment in an intracytoplasmic injection/in-vitro fertilization programme. PMID- 10374091 TI - How long does laparoscopic surgery really take? Lessons learned from 1000 operative laparoscopies. AB - The purpose of this study was to assess the operating time of the most common gynaecological laparoscopic procedures. We analysed retrospectively 1000 consecutive operative laparoscopies on a procedure-by-procedure basis. Diagnostic laparoscopy and laparoscopic sterilization were specifically excluded from the analysis. The various laparoscopic procedures were grouped and analysed under six major categories. The average operating time for all cases was 76.9 min (range 10 400). In 38 cases (3.8%) the laparoscopic procedure was converted to laparotomy. The average operating time for treating ectopic pregnancy and tubal disease was approximately 60 min (range 13-240). Surgery for endometriosis and ovarian cysts averaged 72 min (range 10-240). Laparoscopic myomectomy and hysterectomy averaged 113 and 131 min respectively (range 25-400). Our results show that while the operating time for most operative laparoscopies is less than 75 min, the range of operating times is great. The relative lack of predictability in procedure times means that the efficient utilization of fixed theatre sessions is difficult. PMID- 10374092 TI - Laparoscopic myomectomy in premenopausal women with and without preoperative treatment using gonadotrophin-releasing hormone analogues. AB - The present study was undertaken in order to evaluate the usefulness or otherwise of preoperative gonadotrophin-releasing hormone (GnRH) analogue treatment prior to laparoscopic myomectomy. From June 1993 through December 1996, 60 premenopausal women aged between 25 and 42 years and with a sonographic diagnosis of intramural or subserous myomas were selected for laparoscopic myomectomy at the Department of Obstetrics and Gynaecology of the Catholic University of The Sacred Heart, Rome. According to a computer-generated sequence, 30 patients were submitted to three cycles of GnRH analogue treatment prior to surgery, whereas no preoperative treatment was prescribed to the other 30 patients. Laparoscopic myomectomy was successfully performed in all patients for a total of 174 myomas excised laparoscopically. The patients' mean age, the number of myomas per patient, the mean diameter of the myomas, parity and estimated blood loss were similar in both groups. The operative time was significantly longer in the group of patients submitted to GnRH analogue treatment than that of the group of patients not submitted to any preoperative medical therapy (157.5 +/- 74.71 versus 112.33 +/- 54.71 min; P = 0.01). No intra-operative complications occurred. In no case was blood transfusion necessary. Two patients developed post operative fever (temperature > 38 degrees C.). The mean length of hospital stay was 2.39 days and was similar in both groups. Thirteen spontaneous pregnancies occurred among 24 infertile patients (54.1%). The pregnancy rate for these patients was similar in both groups. The viable term delivery rate was 45.8%. The authors conclude that laparoscopic myomectomy is a feasible and safe procedure. The post-operative pregnancy rate for infertile patients is similar to that following laparotomic myomectomy. The present study suggests that preoperative GnRH analogue treatment does not offer any significant advantages for laparoscopic myomectomy. PMID- 10374093 TI - The role of neutrophils in the formation of peritoneal adhesions. AB - The most common cause of intraperitoneal adhesions which may result in infertility and intestinal obstruction is previous abdominal surgery. Surgical trauma of the peritoneum in the absence of infection elicits a rapid and transient influx of polymorphonuclear leukocytes (PMN) into the peritoneal cavity. The role of neutrophils in intraperitoneal adhesion formation has not been studied. We aimed to study the effects of PMN counts and PMN functions on peritoneal adhesion formation. Forty peritoneal adhesion-induced rats were randomly divided into three groups; group I, receiving saline; group II, receiving cyclophosphamide; and group III, receiving granulocyte-macrophage colony-stimulating factor (GM-CSF). In all groups, peritoneal lavage was performed to determine PMN counts the day after adhesion induction. Blood neutrophil counts and neutrophil functions were also determined. Adhesions were evaluated blindly 14 days after the operation. Adhesion tissue samples were microscopically evaluated. Tissue hydroxyproline and collagen concentrations were measured. The neutrophil counts and phagocytosis significantly increased in group III and neutrophil counts decreased in group II (P < 0.05). The score of adhesion formation in group II was significantly less than that in groups I and III (P < 0.05). Hydroxyproline concentrations of adhesion tissue were significantly decreased in group II when compared with group III (P < 0.05). The present study shows that neutropenia lowers the degree of postoperative adhesion formation. It is concluded that PMN may have a role to play in modulating post-operative adhesion formation. PMID- 10374094 TI - The mouse as a model to study adhesion formation following endoscopic surgery: a preliminary report. AB - Our aim was to investigate the feasibility of a mouse model to study adhesion formation following endoscopic surgery. Following preliminary studies to establish anaesthesia and pneumoperitoneum pressure, a prospective randomized study was carried out to investigate the effect of CO2 pneumoperitoneum on postoperative adhesions. In group I (control group), the duration of pneumoperitoneum was shorter than 5 min. In groups II, III and IV, pneumoperitoneum was maintained for 60 min without flow, with a continuous low flow (1 ml/min) and a continuous high flow (10 ml/min) through the abdominal cavities of the mice using non-humidified CO2, respectively. Adhesions were scored after 7 days by laparotomy. The total adhesion scores were 0.9 +/- 0.8 (n = 15) in control group, 2.4 +/- 0.8 (n = 15) (P < 0.001 versus control group) in group II with no flow, 2.6 +/- 1.3 (n = 15) (P < 0.001 versus control group) in group III with a continuous low flow and 4.3 +/- 0.9 (n = 15) (P < 0.001 versus control group and P < 0.001 versus group II and III) in group IV with a continuous high flow. In conclusion, the mouse can be used as a model to study adhesion formation following endoscopic surgery. Duration of CO2 pneumoperitoneum is a co-factor in adhesion formation. PMID- 10374095 TI - Previously undetected Chlamydia trachomatis infection, immunity to heat shock proteins and tubal occlusion in women undergoing in-vitro fertilization. AB - The relationship between a previously undetected Chlamydia trachomatis infection, tubal infertility, immunity to heat shock proteins and subsequent in-vitro fertilization (IVF) outcome was evaluated. Women with tubal occlusion, with or without hydrosalpinges, and no history of C. trachomatis infection were tested for circulating antibodies to the human 60-kDa heat shock protein (Hhsp60), the C. trachomatis 10-kDa heat shock protein (Chsp10) and C. trachomatis surface antigens prior to their initial IVF cycle. Sera were obtained from 50 women whose male partners were infertile, 58 women with tubal occlusion but no hydrosalpinx and 39 women with tubal occlusions plus hydrosalpinx. Clinical pregnancies were documented in 68% of the women with male factor infertility. This was higher than the 43.1% rate in women with tubal occlusions (P = 0.04) and the 41% rate in women with hydrosalpinx (P = 0.02). C. trachomatis antibodies were present in one (2%) women with male factor infertility as opposed to 15 (25.9%) women with tubal occlusion (P = 0.003) and 13 (33%) with hydrosalpinx (P < 0.0001). Antibodies to Chsp10 were more prevalent in women with hydrosalpinx (46.8%) than in women with male factor infertility (P < 0.0001, 6%) or tubal occlusion (P = 0.0009, 15.5%). Hhsp60 antibodies were equally more prevalent in women with tubal occlusion plus (46.8%) or minus hydrosalpinx (41.4%) than in women with male factor infertility (P < 0.0002). Hhsp60 was more prevalent in those women positive for Chsp10 (P = 0.02) or C. trachomatis (P = 0.04) antibodies than in women lacking these antibodies. There was no relationship between any of the antibodies measured in sera and IVF outcome. PMID- 10374096 TI - Comparison between intracytoplasmic sperm injection and in-vitro fertilization (IVF) with high insemination concentration after total fertilization failure in a previous IVF attempt. AB - The aim of this prospective study was to evaluate whether couples with total fertilization failure in a previous in-vitro fertilization (IVF) attempt should be offered an additional IVF treatment with elevated insemination concentration or should be treated with intracytoplasmic sperm injection (ICSI). In 23 cycles 228 sibling metaphase II (MII) oocytes were randomly divided: 143 and 85 oocytes were utilized for ICSI and IVF respectively. Of the 143 injected (ICSI) oocytes, 90 (62.9%) were normally fertilized (two pronuclei), whereas 21 (14.7%) oocytes were damaged by the ICSI procedure. Of the fertilized oocytes 72 (80%) developed into transferable embryos. No fertilization at all was observed in the 85 sibling MII oocytes which were inseminated (P < 0.001). In all 23 cycles at least one embryo, obtained by ICSI, could be replaced. Eight pregnancies were achieved of which six resulted in the delivery of nine healthy children. In conclusion, for couples with no or almost no fertilization of oocytes in previous IVF attempts, ICSI appeared to be far superior to an additional IVF attempt with further elevated insemination concentrations. PMID- 10374097 TI - Anti-nuclear antibodies in patients with premature ovarian failure. AB - We examined the prevalence of anti-nuclear antibodies (ANA) in 32 consecutive patients with premature ovarian failure with and without chromosomal abnormalities. Blood samples were taken for karyotype determination as well as detection of autoantibodies, X-terminal microdeletions and spontaneous follicular growth. The correlation between ANA positivity and the age at onset of amenorrhoea, as well as the presence of karyotype abnormalities, X-terminal microdeletions and follicular growth was determined. Ten of the 24 patients with normal karyotype and none of the 8 patients with karyotype abnormalities were ANA positive. ANA were found more frequently in patients with premature ovarian failure with normal karyotypes than in control amenorrhoeic patients (42 versus 6, P < 0.01). ANA were found in 77% (10/13) of premature ovarian failure patients with normal karyotypes who developed amenorrhoea at or under the age of 30 years, but not in the patients who developed amenorrhoea later in life. Follicular growth was evident in 50% (5/10) of karyotypically normal patients with ANA, 71% (10/14) of karyotypically normal patients without ANA and 38% (3/8) of patients with karyotype abnormalities. X-terminal microdeletions were not found in any of the patients studied. These results suggest that patients with premature ovarian failure and ANA are an aetiologically and clinically distinct group. PMID- 10374099 TI - A possible effect of different light sources on pregnancy rates following gamete intra-fallopian transfer. AB - A retrospective study of 34 sequential gamete intra-Fallopian transfer (GIFT) procedures suggested a significant effect on pregnancy rates associated with the different laparoscopic light sources, with a pregnancy rate of 50% in 22 cycles using a halogen light source and 9% in 12 cycles using a xenon light source. Other explanatory variables were explored, but none was to have a significant effect on the pregnancy rate. Further investigation revealed that the xenon light source emitted more ultraviolet light than the conventional halogen light source- suggesting a possible detrimental effect of ultraviolet light on the gametes in the GIFT procedure. PMID- 10374098 TI - A prospective randomized study comparing the outcome of in-vitro fertilization and embryo transfer following culture of human embryos individually or in groups before embryo transfer on day 2. AB - A prospective randomized trial of in-vitro fertilization and embryo transfer was undertaken to investigate the reported beneficial effects of culturing preimplantation human embryos in groups, rather than individually. A total of 159 treatment cycles, in which the women were matched for age, basal gonadotrophin concentrations and number of previous attempts, were included in the study. Of these, 78 cycles were randomized to the 'individual culture' group, and 81 cycles were randomized to the 'group culture' group. The groups did not differ in terms of the median number of oocytes or embryos obtained per cycle. There was no statistically significant difference between the two groups in terms of treatment outcome, as assessed by pregnancies or clinical pregnancies. PMID- 10374100 TI - Comparison of two elective transfer policies of two embryos to reduce multiple pregnancies without impairing pregnancy rates. AB - A first elective transfer policy of two embryos based solely on embryo morphology was compared to a more restrictive policy transferring two embryos to all patients aged < 35 years with less than three previous cycles to reduce the incidence of multiple pregnancies. With a significant reduction in the number of triple transfers from 72.4 to 44.3%, the delivery rates were similar for both policies, 31 and 32.1%. However, the multiple pregnancy rates per transfer significantly decreased from 12.5 to 7.8% (P < 0.05). Of 99 pregnancies, only 24.2% were multiple including 1% of triplets compared to 40.7% multiple pregnancies including 6.7% of triplets for the first policy. Forty-eight transfers of two average embryos with the new policy were compared to 264 transfers of three average embryos with the old policy. Multiple pregnancy rates per transfer were significantly reduced by a third from 23 to 8% (P < 0.05) without a reduction of the pregnancy rates (42 and 48%). This study demonstrated that elective transfer of two embryos reduced the number of multiple pregnancies without impairing the pregnancy rates even with the transfer of average embryos. PMID- 10374101 TI - No differences in outcome after intracytoplasmic sperm injection with fresh or with frozen-thawed epididymal spermatozoa. AB - This retrospective consecutive case series aimed at comparing the results of intracytoplasmic sperm injection (ICSI) with fresh and with frozen-thawed epididymal spermatozoa obtained after microsurgical epididymal sperm aspiration (MESA) in 162 couples. These couples were suffering from infertility because of congenital bilateral absence of the vas deferens (n = 109), failed microsurgical reversal for vasectomy or postinfectious epididymal obstruction (n = 44), irreparable epididymal obstruction (n = 4), ejaculatory duct obstruction (n = 2) or anejaculation (n = 3). Overall, 176 MESA procedures were performed in the husbands, followed by 275ICSI procedures with either fresh (n = 157) or frozen thawed (n = 118) epididymal spermatozoa. No significant differences were observed in the parameters of spermatozoa used either freshly or frozen-thawed. In the fresh epididymal sperm group 59.4% of all the injected oocytes fertilized normally as compared to 56.2% of all injected oocytes in the frozen-thawed epididymal sperm group, and embryonic development was comparable between the two groups. A total of 245 transfers were performed: 145 after the use of fresh epididymal spermatozoa and 100 after the use of frozen-thawed spermatozoa. The overall pregnancy rate per ICSI cycle was significantly lower when frozen-thawed epididymal spermatozoa were used (26.3 versus 39.5%). However, no significant differences were found either in clinical and ongoing pregnancy rates or in implantation rates. There were no differences in pregnancy outcome. In patients suspected of having obstructive azoospermia with no work-up or an incomplete one, MESA is the preferred method for sperm recovery because a full scrotal exploration can be performed and, whenever indicated, a vasoepididymostomy may be performed concomitantly. Recovery of epididymal spermatozoa for cryopreservation during a diagnostic procedure is certainly a valid option in these patients since ICSI may be performed later or even in another centre using the frozen-thawed epididymal spermatozoa without jeopardizing the ICSI success rate. PMID- 10374102 TI - Anovulations in an ovary during two menstrual cycles enhance the pregnancy potential of oocytes matured in that ovary during the following third cycle. AB - The aim of this study was to test whether ovulation from an ovary affects the health of oocytes from dominant follicles in that ovary two cycles later. A total of 80 women each with two intact ovaries underwent 270 treatment cycles (155 natural cycles and 115 clomiphene citrate cycles) all showing unilateral ovulation. The results from the in-vitro fertilization (IVF) treatment were grouped according to whether ovulation (O) or anovulation (A) (no ovulation) was observed in the ovary with dominant follicle during the treatment cycle in the previous two cycles: O-O, A-O, O-A and A-A (previous second cycle-previous first cycle). The rate of pre-embryo formation in A-A was significantly higher than that of O-A. The pregnancy rate in A-A (29%) was also higher than those of O-A (13%), A-O (9%) and O-O (5%). These rates increased from O-O to A-A as the number of previous ovulations in an ovary decreased. The presence of a corpus luteum and/or a dominant follicle is likely to exert local negative effects on the health of the oocyte contained in the follicle selected to ovulate up to two cycles later. Anovulations in an ovary for two menstrual cycles may therefore provide improved conditions for the development of a healthier oocyte with an increased pregnancy potential. PMID- 10374103 TI - Delivery rates after in-vitro fertilization following bilateral salpingectomy due to hydrosalpinges: a case control study. AB - This retrospective case-control study assessed the impact of bilateral salpingectomy due to uni- or bilateral hydrosalpinges on the outcome of in-vitro fertilization (IVF) in a large consecutive series of patients. The effect of bilateral salpingectomy due to hydrosalpinges on pregnancy outcome was compared in 139 patients (263 cycles) and 139 age-matched controls with tubal infertility without hydrosalpinges (296 cycles). The delivery rates per initiated cycle as well as the implantation rates were equal in the two groups (21.7 versus 21.6% and 19 versus 21%). The number of embryos, the cleavage stage, and the embryo morphology score were equal in the two groups. Among 92 patients treated with 182 IVF cycles who underwent salpingectomy between 1.5 and 5 years prior to their first IVF cycle, the delivery and the implantation rates were 22.5 and 20.5% respectively. Of the patients with salpingectomy after an average of 1.7 failed IVF cycles and who re-entered the IVF programme 3 and 6 months subsequent to surgery, 47 were treated with 83 IVF cycles. The live birth and the implantation rates after surgery in this group were 20.5 and 20% respectively. It is concluded that bilateral salpingectomy due to hydrosalpinges restores a normal delivery as well as implantation rate after IVF treatment compared to controls. A favourable outcome is also found in patients operated on after repeated IVF failures. Furthermore, a normal live birth rate as well as a high implantation rate is maintained for at least three IVF cycles subsequent to surgical treatment. PMID- 10374104 TI - Diurnal variation of semen quality in human males. AB - The possibility of a diurnal variation in semen quality was tested in 54 human males attending our infertility clinic. Of the enrolled subjects, 24 were normozoospermic and 30 were suffering from oligo- and/or asthenozoospermia. Seminal fluid was collected by masturbation twice by each subject, once in the morning (7:00-7:30 a.m.) and once in the afternoon (5:00-5:30 p.m.). Abstinence from sexual intercourse for 3-4 days before each of the two collections was requested. Semen parameters were evaluated independently by two biologists before and after pellet swim-up. Beside similar macroscopic parameters, specimens collected in the afternoon showed a higher number (P < 0.01) and concentration (P < 0.01) of spermatozoa. Also, immediately (P < 0.05), and at 1 h (P < 0.02) and 2 h (P < 0.01) after pellet swim-up, the number of spermatozoa with progressive linear motility was higher in the afternoon than in the morning. These data are the first documenting a diurnal rhythm in sperm quality which may contribute to the reported variability in semen parameters, and may prove useful for spontaneous and assisted conceptions. PMID- 10374105 TI - Assisted reproduction for infertile patients with 9 + 0 immotile spermatozoa associated with autosomal dominant polycystic kidney disease. AB - We investigated the clinical feature of patients with totally immotile spermatozoa due to 9 + 0 ultrastructural flagellar defects and polycystic kidney disease. We also tried to establish the feasibility of applying modern assisted reproduction technology (ART) in these patients. During 6-year interval a total of 1956 Japanese men were referred to the male infertility clinic. Of them, 16 were diagnosed to have immotile spermatozoa and four of them exhibited axonemal 9 + 0 defects in the sperm flagella. These four also had autosomal dominant polycystic kidney disease (ADPKD). Intrauterine insemination (IUI) and conventional in-vitro fertilization and embryo transfer failed to achieve fertilization. Intracytoplasmic sperm injection (ICSI) with 100% immotile spermatozoa was performed in all four cases. Two-pronuclear fertilization was obtained in 27 of the 70 (38.6%) of the successfully injected oocytes, but no pregnancy resulted. In one case, a few motile spermatozoa were present at the second cycle of ICSI, a pregnancy was successfully achieved using these spermatozoa. While immotile spermatozoa from patients with the axonemal 9 + 0 defect achieved fertilization by ICSI, the embryos failed to develop. Our results indicate that the central microtubules may play a role in fetal development. Since the 4 patients with 9 + 0 defects also had ADPKD, the genetic linkage between these two conditions should be studied by molecular biological methods so as to aid our ability to counsel such patients. PMID- 10374107 TI - Diagnostic epididymal and testicular sperm recovery and genetic aspects in azoospermic men. AB - Various procedures for sperm recovery in azoospermic men have been described, from open testicular biopsy to simple needle aspiration from the epididymis and the testis. Fifty-one obstructive and 86 non-obstructive azoospermic men were treated to compare the recovery of spermatozoa obtained by percutaneous aspiration from the epididymis (PESA) and aspiration/extraction from the testis (TESA, TESE) with histopathology. If TESA failed, the work up proceeded with TESE. All patients were karyotyped. Spermatozoa were recovered by PESA or TESA in all obstructive men (51/51 patients). In 22 out of 86 patients with non obstructive azoospermia, testicular spermatozoa could be successfully recovered by TESA. In five additional patients TESE was successful in recovering spermatozoa where TESA had failed. In 43 patients, neither TESA nor TESE was successful. Sixteen patients chose not to proceed with TESE. Seven out of 86 patients had an abnormal karyotype in the non-obstructive group (8%), none in the obstructive group. In the non-obstructive patient group testicular histopathology showed hypospermatogenesis, incomplete maturation arrest and germ cell aplasia with focal spermatogenesis in cases where spermatozoa were recovered and complete germ cell aplasia, complete maturation arrest and fibrosis in cases where no spermatozoa were found. Spermatozoa were recovered by PESA or TESA from all patients with obstructive azoospermia and from approximately 40% of patients with non-obstructive azoospermia by TESA or TESE. Retrieval of viable spermatozoa in the infertility work-up was highly predictable for sperm recovery in subsequent ICSI cycles. TESA performed under local anaesthesia seems almost as effective as more invasive procedures in recovering testicular spermatozoa, both in obstructive and non-obstructive azoospermic men. PMID- 10374106 TI - Pre-freezing sperm preparation does not impair thawed spermatozoa binding to the zona pellucida. AB - The present study was conducted to assess the fertilizing potential of frozen thawed spermatozoa, which were cryopreserved after separation on a Percoll gradient, or washed out of seminal plasma. For this purpose, binding to the zona pellucida and other characteristics of the treated sperm cells were compared with those of cryopreserved spermatozoa from the same original sample which were not manipulated before freezing. Semen specimens were obtained from 80 candidates for sperm donation. Percoll-treated sperm samples compared with the sibling, unprocessed controls had significantly higher values of sperm motility characteristics and per cent of cells with normal morphology after freezing and thawing. Sperm binding ability to the zona pellucida was not statistically different (109 +/- 8.1% and 94 +/- 6.7% in unprocessed and Percoll-treated samples respectively). Sperm specimens processed by washing had significantly higher values for motility characteristics than untreated sibling samples, but no differences were found between the treated and untreated samples for morphology and binding to the zona pellucida (hemizona index of 75 +/- 7.0% and 76 +/- 6.7% in unprocessed and washed samples respectively). These findings suggest that, judged by the binding assay, the aforementioned pre-freezing separation processes have no adverse effect upon the fertilizing potential of the thawed sperm cells. These procedures make it possible to optimize the progressive motile sperm cell concentration of the frozen specimen, which facilitates the storage of samples with good quality, even when the features of the original semen are sub-optimal. PMID- 10374108 TI - Morphology comparison of individually selected hyperactivated and non hyperactivated human spermatozoa. AB - The objective of this study was to compare the morphology of human spermatozoa undergoing hyperactivated motility in vitro with those that were non hyperactivated (non-hyp). Hyperactivation criteria were established by the Hobson Sperm Tracker (HST), sampling at 25 Hz, as curvilinear velocity (VCL) > or = 70 microns/s, amplitude of lateral head displacement (ALH) > or = 7 microns, linearity (LIN) < or = 30% and straight-line velocity (VSL) < or = 30 microns/s. Specially developed software incorporated in the HST produced a white computer generated overlay for spermatozoa satisfying hyperactivation criteria. These spermatozoa, visually identified on a tracking monitor, were individually removed with micromanipulation equipment using a 12 microns-diameter needle. Fifty-six patient ejaculates were examined comprising a total morphological analysis of 1886 non-hyp spermatozoa and 1051 hyperactivated spermatozoa. Hyperactivated spermatozoa had a significantly higher mean percentage of normal heads and small acrosomes (P < 0.0001 and < 0.0001 respectively) and a significantly lower percentage of large and round heads, midpieces and tail defects (P = 0.002, < 0.0001, 0.02 and < 0.0001 respectively) when compared with non-hyp spermatozoa. These data demonstrate, for the first time, that a homogeneous live population of human hyperactivated spermatozoa, selected in vitro from patients with highly variable degrees of teratozoospermia, is comprised predominantly of cells with normal morphology (P < 0.0001). PMID- 10374109 TI - Testicular sperm extraction: microdissection improves sperm yield with minimal tissue excision. AB - Testicular sperm extraction (TESE) is often an effective method for sperm retrieval from men with non-obstructive azoospermia. However, TESE has been a blind procedure that does not identify the focal sperm-producing areas of the testicle until after tissue has been excised from the patient. Experience with a new technique of microdissection of testicular tubules is presented here that identifies sperm-containing regions before their removal. Identification of spermatogenically active regions of the testicle is possible by direct examination of the individual seminiferous tubules. The underlying concept for this technique is simple: seminiferous tubules containing many developing germ cells, rather than Sertoli cells alone, are likely to be larger and more opaque than tubules without sperm production. In a sequential series of TESE cases for men with non-obstructive azoospermia, the ability to find spermatozoa increased from 45% (10/22) to 63% (17/27) after introduction of the microdissection technique. Microdissected samples yielded an average of 160,000 spermatozoa per sample in only 9.4 mg of tissue, whereas only 64,000 spermatozoa were found in standard biopsy samples that averaged 720 mg in weight (P < 0.05 for all comparisons). For men where microdissection was attempted, successful identification of enlarged tubules was possible in 56% (15/27) of cases. However, spermatozoa were retrieved with microdissection TESE for six men in whom sperm retrieval was unsuccessful with standard TESE approaches (35% of all men with spermatozoa retrieved). These findings suggest that microdissection TESE can improve sperm retrieval for men with non-obstructive azoospermia over that achieved with previously described biopsy techniques. PMID- 10374111 TI - Germ cell transfer into rat, bovine, monkey and human testes. AB - Germ cell transplantation is a potentially valuable technique offering oncological patients gonadal protection by reinitiating spermatogenesis from stem cells which were reinfused into the seminiferous tubules. In order to achieve an intratubular germ cell transfer, intratubular microinjection, efferent duct injections and rete testis injections were applied on dissected testes of four different species: rat, bull, monkey and man. Ultrasound-guided intratesticular rete testis injection was the best and least invasive injection technique with maximal infusion efficiency for larger testes. Deep infiltration of seminiferous tubules was only achieved in immature or partially regressed testes. This technique was applied in vivo on two cynomolgus monkeys. In the first monkey a deep infusion of injected cells and dye into the lumen of the seminiferous tubules was achieved. In the second, transplanted germ cells were present in the seminiferous epithelium 4 weeks after the transfer. These cells were morphologically identified as B-spermatogonia and located at the base of the seminiferous epithelium. In summary, this paper describes a promising approach for germ cell infusion into large testes. The application of this technique is the first successful attempt of a germ cell transfer in a primate. PMID- 10374110 TI - Reversion of the differentiated phenotype and maturation block in Sertoli cells in pathological human testis. AB - To study the relationship between abnormal Sertoli cell differentiation and spermatogenic impairment, we examined the expression of Sertoli cell markers normally lost at puberty, cytokeratin 18 (CK18), anti-Mullerian hormone (AMH) and M2A antigen, in three children (aged 1-2 years), 50 adults (aged 19-45 years) with obstructive or non-obstructive azoospermia or oligozoospermia, and six patients (aged 1-18 years) with 5 alpha-reductase deficiency. There was CK18 and/or AMH expression, but never M2A antigen expression, associated with spermatogonial arrest or Sertoli cell-only (SCO) syndrome in infertile men. Loss of M2A antigen suggests the transition of Sertoli cells to an adult phenotype, while CK18 and/or AMH expression may be a manifestation of de-differentiation of Sertoli cells. In 5 alpha-reductase deficiency, there was a sequential loss of CK18, M2A antigen and AMH around puberty, associated with partial spermatogenesis. The persistence of immature Sertoli cells expressing M2A antigen was associated with prepubertal seminiferous cords and SCO syndrome. Therefore, 5 alpha-reductase deficiency may prevent the maturation of Sertoli cells, resulting in impairment of spermatogenesis, and loss of M2A antigen expression coincides with a critical step in the Sertoli cell maturation. High follicle stimulating hormone concentrations due to failure of normal Sertoli cell differentiation indicate a normal development pattern of the hypothalamic-pituitary-gonadal axis. PMID- 10374112 TI - The action of vasoconstrictive agents on human tubal arteries. AB - The aim of this investigation was to compare the response of small arteries of the human tubo-ovarian vasculature to certain vasoactive agents. Ring preparations of the arteries were isolated and mounted in tissue chambers for isometric recording of wall tension. The arteries were exposed to the vasoactive agents adrenalin, prostaglandin F2 alpha and two vasopressin analogues. Adrenalin, prostaglandin F2 alpha, lysin-vasopressin and triglycyl-lysine vasopressin all produced powerful vasoconstriction, the greatest efficacy being shown by and lysine-vasopressin. The maximum response occurred after addition of a third compound to a combination of two, irrespective of which combination was used. Adrenalin showed faster contraction velocity than the other agents. The results indicate that the human tubo-ovarian arteries may be constricted by a variety of physiological and pharmacological stimuli, at least partly acting via different effector mechanisms. It is proposed that these vasoconstrictive agents- alone or in combination--may be useful in conjunction with gynaecological endoscopic surgery, e.g. in tubal pregnancy or ovarian cysts. PMID- 10374113 TI - Are there any relationships between the fecundity of bilateral ovaries in an individual patient and the incidence of apoptotic granulosa cells? AB - Many researchers have discussed fecundity on a per patient or per ovarian follicle basis. In contrast, this study was undertaken on a per ovary basis, and tests the hypothesis that there is a relationship between the fecundity of the bilateral ovaries in an individual patient and the incidence of apoptotic granulosa cells. The two ovaries of 10 women undergoing ovulation induction for in-vitro fertilization (IVF) with gonadotrophin-releasing hormone analogue (GnRHa) and gonadotrophins were compared. There was no difference in apoptotic index in granulosa cells and various hormones in the follicular fluids. Our results indicate that, in the ovulation induction protocol for IVF, GnRHa and gonadotrophins, there is no predisposition of one ovary over the other in an individual patient in terms of apoptosis at the time of aspiration, even though the number of oocytes in each ovary is different, because the number of oocytes retrieved may reveal ovarian fecundity before stimulation with GnRHa + human menopausal gonadotrophin (HMG). PMID- 10374114 TI - Cytokines in the follicular fluid of stimulated and non-stimulated human ovaries; is ovulation a suppressed inflammatory reaction? AB - We determined the concentrations of tumour necrosis factor (TNF)-alpha, interleukins (IL)-1 beta, -6, -8 and -1-receptor antagonist (IL-1-ra) and of oestradiol and progesterone in the follicular fluid of 111 women undergoing in vitro fertilization (IVF) and of six women with ovarian cysts in order to elucidate mid-cycle mechanisms causing dissociation of the follicle wall and local rupture of the ovarian tissue complex. Four stimulation protocols were administered: gonadotrophin releasing hormone agonist/human menopausal gonadotrophin (GnRHa/HMG), clomiphene citrate/HMG (CC/HMG), HMG and follicle stimulating hormone (FSH). Concentrations of TNF alpha and IL-1 beta were below 15 and 3 pg/ml respectively. IL-6 (median 4.1, 3.5-4.4 pg/ml, 95% CI) was higher after stimulation with FSH (5.6 pg/ml) than with HMG (3.2 pg/ml, P < 0.05) or GnRHa/HMG (3.7 pg/ml, P < 0.05), and after stimulation with CC/HMG (5.5 pg/ml) than with HMG (P < 0.01) or GnRHa/HMG (P < 0.001). IL-8 ranged from 32 to 1241 pg/ml (147, 117-178 pg/ml) and IL-1-ra from < 31 to > 10,000 pg/ml (156, 109-192 pg/ml). Cytokine levels did not correlate to oestradiol or progesterone concentrations. The ovarian cysts contained similar IL-8 (14-540 pg/ml) and IL-1 beta (< 30 pg/ml), but higher IL-6 (13.6-> 500 pg/ml) and lower IL-1-ra concentrations. We assume that IL-6, IL-8 and IL-1-ra are involved in peri ovulatory cellular interactions. Thus, ovulation appears to be a cytokine regulated process of an 'inflammation' (IL-6 and IL-8) followed by 'anti inflammatory' reactions (IL-1-ra). PMID- 10374115 TI - The outcome of in-vitro fertilization treatment in women with sonographic evidence of polycystic ovarian morphology. AB - This study compared the outcome of a course of up to three cycles of in-vitro fertilization (IVF) treatment in 46 women (97 cycles) who had polycystic ovaries (PCO) seen on ultrasound scan, but who had no clinical symptomatology associated with polycystic ovary syndrome, with that of 145 women (332 cycles) who had normal ovarian morphology on ultrasound examination. All 191 women had normal early follicular phase serum follicle stimulating hormone (FSH) concentrations, were less than 40 years of age and used the long protocol of pituitary suppression with gonadotrophin-releasing hormone agonist therapy. On average, the women with PCO produced more follicles, oocytes and embryos than the women with normal ovaries, but the fertilization, cleavage and miscarriage rates were similar. Adjusted for age, the odds of achieving a pregnancy within three cycles of treatment in a woman with PCO were 69% higher than those of a woman with normal ovaries [odds ratio (OR): 1.69, 95% confidence interval (CI) 0.99-2.90, P = 0.05)] and the odds of achieving a live birth were 82% higher (OR: 1.82, 95% CI 1.05-3.16, P = 0.03). There is, therefore, evidence that outcome of IVF treatment for women with PCO seen on ultrasound examination may be better than that for women with normal ovaries. PMID- 10374116 TI - Immunocytogenetic detection of normal and abnormal oocytes in human fetal ovarian tissue in culture. AB - This study aimed to: (i) determine whether oocytes are present in cultures of human fetal ovary; (ii) identify whether meiotic anomalies are evident; and (iii) assess whether preparation or culture conditions influence oocyte survival and meiotic progression. Ovaries were collected from fetuses after termination at 13 16 weeks. Oocyte assessment utilized antibodies specific for synaptonemal complex proteins (associated with chromosomes only during meiosis), and antibodies to centromeric proteins. Fragments of tissue were cultured in minimal essential medium + 10% serum +/- follicle stimulating hormone (100 mIU/ml). The sera were fetal calf serum (FCS), FCS for embryonic stem cells (ES-FCS) and human female serum. The numbers and stages of oocytes were assessed after 7-40 days, and particular arrangements of chromosome synapsis identified. Results in fresh tissue included oocytes at leptotene, zygotene, pachytene and diplotene in three of five samples. Four specimens remained viable in vitro, and three had detectable oocytes after culture. The numbers of oocytes and the proportions of zygotene and pachytene cells increased with time in culture. The proportion of degenerate cells in culture was initially higher than in fresh samples, but declined subsequently. More oocytes were detected in ES-FCS and human serum than in FCS. We conclude that human oocytes survive in culture and that progression through prophase I continues. PMID- 10374117 TI - The effect of pituitary desensitization on ovarian volume measurements prior to in-vitro fertilization. AB - The measurement of ovarian volume has been recently shown to predict follicular response in in-vitro fertilization (IVF), specifically a lower number of retrieved oocytes with decreasing ovarian volume. This test appears to be better than basal follicle-stimulating hormone (FSH) as a prognostic measure of ovarian reserve. However, the effect of pituitary desensitization on ovarian volume has not been previously investigated. We prospectively evaluated 38 women undergoing IVF using a long luteal leuprolide acetate (LA) protocol. All women had their ovarian volume measurements performed on day 21, the day of LA start, and again on the day of gonadotrophin start. The mean age was 30.6 +/- 3.9 years (range 23 37). Basal FSH was 5.4 +/- 1.9 IU/l (range 1.2-10.2). The mean preLA ovarian volume was 7.0 +/- 3.6 cm3 (left ovary 6.8 +/- 3.9, right ovary 7.1 +/- 3.8), compared to 6.3 +/- 4.2 cm3 postLA (left ovary 6.0 +/- 4.9, right ovary 6.5 +/- 4.8) (not significant). The mean number of small antral follicles noted in both ovaries was also unchanged after pituitary desensitization. Pituitary desensitization using LA had no effect on overall ovarian volume measurements. The total number of retrieved oocytes decreased with increasing age and decreasing ovarian volume. PMID- 10374119 TI - The cyclic pattern of the immunocytochemical expression of oestrogen and progesterone receptors in human myometrial and endometrial layers: characterization of the endometrial-subendometrial unit. AB - Immunocytochemistry of oestrogen receptor (ER) and progesterone receptor (PR) expression of the whole uterine muscular wall and the endometrium was performed in order to obtain morphological and functional insights into the regulation of cyclic uterine peristalsis, which is confined to the endometrium and the subendometrial myometrium and serves functions such as rapid and sustained sperm transport. The study revealed that the subendometrial myometrium or stratum subvasculare with a predominantly circular arrangement of muscular fibres exhibits a cyclic pattern of ER and PR expression that parallels that of the endometrium, whereas the outer portion of the uterine wall composed of the stratum vasculare and supravasculare, which represents the bulk of the uterine musculature, does not exhibit a cyclic pattern of ER and PR expression. According to ontogenetic and phylogenetic data from the literature, the outer myometrium is of non-paramesonephric origin with functions confined to parturition, while the inner myometrial layer together with the glandular epithelium and the stroma of the endometrium is of paramesonephric origin with various functions during the cycle in addition to those during pregnancy and parturition. The inner quarter of the stratum vasculare adjacent to the stratum subvasculare constitutes a transitional zone in that the cyclicity of receptor staining becomes, in radial direction, gradually less expressed. Morphologically this zone corresponds to the inner part of the stratum vasculare where its muscular fibres blend with those of the stratum subvasculare. PMID- 10374118 TI - Menorrhagia and uterine artery blood flow. AB - Menorrhagia is a significant problem in women of reproductive age. In half of the cases no specific aetiology is known. Vascular factors play a role but remain poorly understood. We chose to study whether any association exists between the flow impedance of uterine arteries and the amount of menstrual blood loss. The study population consisted of 60 spontaneously menstruating 35- to 49-year-old women without endometrial hyperplasia, polyps, or submucous fibroids. The pulsatility index (PI) from uterine arteries, arcuate arteries, and radial arteries was measured by transvaginal colour Doppler. Menstrual blood loss was measured by the alkaline haematin method. A significant inverse correlation was found between uterine artery PI and the amount of menstrual blood loss, suggesting that women with lower uterine flow impedance bleed more. A regression model confirmed that this association was specific and not explained by uterine size, fibroids or any other of the 11 potential confounders included in the model. The correlation between uterine artery PI and amount of menstrual blood loss suggests that vascular factors may be involved in the pathogenesis of menorrhagia. PMID- 10374120 TI - Disparate actions of mifepristone (RU 486) on glands and stroma in the primate endometrium. AB - Besides being an antiprogestin, mifepristone (RU 486) was recently shown to antagonize oestrogen-dependent growth in the endometrium. To explore the molecular mechanisms for this phenomenon, we investigated whether or not the morphological effects of mifepristone are mediated by the progesterone receptor (PR) and whether mifepristone has disparate effects on the glandular epithelium and stroma. Six groups of hypogonadal, oestrogen-primed cynomolgus monkeys were treated for 2 weeks with: vehicle only (group I); mifepristone (group II); mifepristone plus progesterone at 0.2 mg/kg (group III), 1.0 mg/kg (group IV) or 5.0 mg/kg (group V); and progesterone only (5.0 mg/kg) (group VI). Histomorphological evaluation showed strikingly compacted stroma in the mifepristone-exposed endometria (group II), which was partially reversible by additional progesterone treatment (groups III-V). Glandular proliferation (pseudostratification, glandular mitoses) in mifepristone-treated monkeys was not significantly different from that in vehicle (oestradiol)-treated monkeys, but was inhibited by progesterone-only treatment. Cells containing vacuoles were scarce in the mifepristone-exposed endometrium, but detected frequently in progesterone-exposed endometria, indicating the strong antisecretory effect of mifepristone on glands. We conclude that oestrogen-dependent oedema in the stroma is antagonized by mifepristone. The reversal of this effect by progesterone suggests a PR-mediated mechanism. In glands, mifepristone is antiprogestogenic, but not antioestrogenic. Thus, stromal cells may be the target of antiprogestin induced inhibition of oedema and endometrial growth. PMID- 10374121 TI - Effect of 6 beta-methylprednisolone on mouse pregnancy rate. AB - The aim of this study was to evaluate the effect of methyl-prednisolone on the pregnancy rate in mice. For this reason, zona pellucida-intact and zona pellucida free embryos at the blastocyst stage were transferred to recipient mice at day 2.5 of pseudopregnancy. Embryo transfer was performed into non-immunodepressed and immunodepressed groups of recipient mice using 0.3 or 0.6 microgram/g of 6 beta-methylprednisolone. A higher implantation and developmental rate of zona free embryos transferred to the immunodepressed group of recipients was observed after using the higher dose of methylprednisolone. PMID- 10374122 TI - Haemostatic and metabolic abnormalities in women with unexplained recurrent abortion. AB - The objective of this study was to establish whether or not patients with unexplained recurrent abortion have an increased incidence of haemostatic or metabolic abnormalities. Fifty-two patients with a history of unexplained habitual abortion (two or more spontaneous abortions before 16 weeks' gestation) were tested for protein S, protein C and antithrombin (AT) III deficiency, activated protein C (aPC) resistance, hyperhomocysteinaemia and anticardiolipin antibodies (ACA). The control group consisted of 67 healthy women with a history of only uncomplicated pregnancies. Blood samples were taken for measuring protein S, protein C, AT III, ACA and activated protein C resistance and a methionine loading test was performed. Of the 46 patients tested for protein S deficiency, 8 (17.4%) were positive. Of the 43 patients tested, two (4.7%) were protein C deficient and none was AT III deficient. Of the 42 patients tested for ACA, eight (19.1%) had detectable antibodies. Of the 44 patients tested for aPC resistance, two (4.6%) were positive. Finally, 35 patients were tested for hyperhomocysteinaemia and six (17.1%) were positive. It was concluded that parous women with a history of unexplained recurrent abortion have an increased incidence of hyperhomocysteinaemia and a trend of increased incidence of ACA can be found. PMID- 10374123 TI - Development of the fetal uterus between 19 and 38 weeks of gestation: in-utero ultrasonographic measurements. AB - In-utero assessment of the internal female genitalia is important for determination of fetal gender in fetuses with suspected genital tract anomalies. We therefore measured fetal uterine transverse width and circumference from 19 weeks of gestation until term, using transvaginal and transabdominal high resolution ultrasound techniques in order to establish nomograms. A prospective, cross-sectional study on 180 normal singleton pregnancies was performed. Data were obtained for 140 normal fetuses. The mean +/- SD uterine width and circumference were 12.9 +/- 4.1 mm (95% confidence interval 12.1-13.7), and 40.2 +/- 12.5 mm (95% confidence interval 37.9-42.5) respectively. Uterine size as a function of gestational age was expressed by the regression equations: uterine width (mm) = 12.9 + 0.7 x gestational age (weeks), and uterine circumference (mm) = 40.2 + 2.1 x gestational age. The correlation coefficients, r = 0.885 and r = 0.888, for uterine width and circumference, by gestational age respectively, were highly statistically significant (P < 0.001). A nomogram of uterine width and circumference per gestational week, and the 95% prediction limits were defined. The present data offer baseline measurements of the fetal uterus that may allow intrauterine assessment of the female genital tract and associated fetal gender. PMID- 10374124 TI - Gestational hypertension but not pre-eclampsia is associated with insulin resistance syndrome characteristics. AB - The aim of this study was to assess whether the metabolic characteristics of insulin resistance syndrome are present in pre-eclamptic (PE), gestational (GH) and chronic hypertensive (CH) pregnancies. Glucose and insulin serum concentrations, both fasting and after oral administration of a glucose tolerance test, were evaluated in 26 hypertensive pregnant women (10 PE, 10 GH and six CH patients) and in 10 healthy controls during the third trimester of gestation. Insulin sensitivity was assessed using the hyperinsulinaemic-euglycaemic clamp technique. The plasma concentrations of triglyceride (TG), high density (HDL), low density (LDL), and very low density (VLDL) lipoprotein cholesterol, apolipoproteins AI and B, and non-esterified fatty acid (NEFA) were also measured. Women with GH exhibited approximately 40% lower steady-state insulin sensitivity index (ISI) compared to controls (3.75 versus 6.34, P < 0.03), as well as approximately 33% higher mean plasma TG (3.57 versus 2.68 mmol/l, P < 0.01), and approximately 69% higher mean NEFA (0.59 versus 0.35 mmol/l, P < 0.01). Women with PE showed similar ISI but reduced insulin and glucose areas under curve compared to controls (P < 0.006, P < 0.0005 respectively). Women with PE also had higher HDL-cholesterol and apo-AI than controls. Patients with CH had similar lipid and carbohydrate metabolism to control subjects. In conclusion, women with GH exhibit metabolic features similar to those of patients with insulin resistance syndrome, suggesting that similar abnormalities could be involved in the pathogenesis of these disorders. In contrast, our data do not support an association between insulin resistance syndrome and hypertension in pregnant women with PE and chronic hypertension. PMID- 10374125 TI - Marked variation in antiphospholipid antibodies during pregnancy: relationships to pregnancy outcome. AB - Variations in blood concentrations of antiphospholipid antibodies (APA) were investigated through the course of pregnancy in women who had a history of recurrent pregnancy loss, and were related to changes in pregnancy outcome. Serial measurements of APA were made in 16 women with antiphospholipid syndrome (APS) and 16 with negative APA tests pre-pregnancy. There was considerable intraindividual variation in test results through pregnancy. There was a significantly higher ratio of dilute Russell's viper venom time and IgG ACA titre in the first trimester compared with results pre-pregnancy in women with APS. Furthermore, transiently positive APA results were noted in the control group during pregnancy and some women with antiphospholipid syndrome tested negative for APA in mid- and late pregnancy. We have demonstrated clinically important variations in the results of tests for APA during pregnancy in women with APS. PMID- 10374126 TI - Leukocytes infiltrate the myometrium during human parturition: further evidence that labour is an inflammatory process. AB - Inflammatory mediators in the cervix, placenta and fetal membranes play a crucial role in human parturition. The aim of this study was to determine whether the upper and lower segments of the myometrium are infiltrated by inflammatory cells during pregnancy and parturition. Myometrial biopsies were obtained from non pregnant women, and pregnant women at term before and after the onset of spontaneous labour. Subpopulations of inflammatory cells were identified using immunocytochemistry. The intercellular adhesion molecules, 1 and 2, platelet endothelial cell adhesion molecule, vascular cell adhesion molecule and E selectin were immunolocalized to investigate their involvement in leukocyte accumulation. Histological analysis demonstrated that inflammatory cells, predominantly neutrophils and macrophages, infiltrate human myometrium during spontaneous labour at term. The infiltrate is predominant in the lower uterine segment but is also present in the upper segment. Increased expression of E selectin was found on the vascular endothelium of biopsies obtained during labour, suggesting a role for this molecule in the accumulation of leukocytes. These results suggest that inflammatory cell infiltration is part of the physiological mechanisms that occur in the myometrium during parturition. Further understanding of this process may suggest new strategies aimed at preventing preterm delivery. PMID- 10374127 TI - Structural characteristics of term human fetal membranes prior to labour: identification of an area of altered morphology overlying the cervix. AB - Premature rupture of fetal membranes can have serious clinical implications, especially for the initiation of preterm labour and its consequences. To account for this phenomenon many studies have attempted to identify membrane features that may be uniquely associated with the site of rupture. Our previous work has identified an area of the fetal membrane, following spontaneous term birth which exhibits alterations consistent with structural weakness. The aim of this study was to determine if these changes existed prior to labour. In formalin-fixed paraffin-embedded tissue sections an area of the fetal membrane overlying the cervix, termed the 'cervical membranes', was characterized by an increased thickness of the connective tissue layer (215% increase, P < 0.01) and decreased thickness of both the cytotrophoblast (36% reduction, P < 0.01) and decidual layers (64% reduction, P < 0.01) compared to the rest of the membrane. This resulted in the cervical membranes being significantly thinner (P < 0.05) than the rest of the membrane. Similar changes were also detected in frozen sections of fetal membranes. These regional differences have two important implications in that: (i) the cervical membrane may represent a region of structural weakness susceptible to rupture during labour, and (ii) the paracrine relationships between fetal membranes and the myometrium may be qualitatively affected within different regions of the uterus. PMID- 10374128 TI - Mother-to-child transmission of human immunodeficiency virus in Italy: temporal trends and determinants of infection. The Italian Collaborative Study on HIV infection in pregnancy. AB - In order to analyse temporal trends in vertical transmission rates of human immunodeficiency virus (HIV) and determinant of congenital HIV infection in Italy, we have considered data from a network of hospitals co-operating in the Italian Collaborative Study on HIV infection in pregnancy, conducted between 1988 and 1995. A total of 1040 women entered the study. The HIV-1 status of the babies was known in 848 cases (81.5%). Transmission rates were highest in the period 1988-1991, then tended to decrease and in 1995 the rate was 9.7 per 100 children (this finding, however, was based on only six infected children and the trend was not statistically significant). Considering the overall series, the risk of vertical HIV transmission was higher in women with low CD4 count in pregnancy [odds ratio (OR) < 400 versus > or = 400 1.8, 95% confidence interval (CI) 1.1 2.9]. In comparison with vaginal delivery the risk of transmission was 0.3 (95% CI 0.1-0.5) and 0.6 (95% CI 0.3-1.2) respectively for elective and emergency delivery. In comparison with women who delivered at term (> or = 37 gestation weeks) the OR of HIV infection of the babies for the whole series was 2.2 (95% CI 1.3-3.6) in women who delivered preterm. Similar findings emerged when the analysis was conducted considering, separately, subjects observed in the period 1988-1991 and 1992-1995. The frequency of Caesarean section increased from 26.5% of deliveries in 1988-1991 to 36.2% in 1992-1995. Consequently, most temporal differences disappeared after standardization for mode of delivery, but the rate in 1995 was still lower than in 1988-1994. PMID- 10374129 TI - Absence of chromosomal instability in spermatozoa of men affected by testicular cancer. AB - Testicular germ cell cancer affects mainly young men. It is the most frequent type of cancer in 20-35 year old men. Since cancer treatment using antineoplasic drugs and ionizing radiation has a negative effect on the function of the gonads, testicular cancer patients are offered the opportunity to cryopreserve their semen samples before the beginning of therapy. For this reason it would be of interest to know whether there is chromosome instability in their spermatozoa prior to any treatment. Using the interspecific human-hamster fertilization system, we have analysed a total of 340 chromosome complements from spermatozoa of control donors and 320 chromosome complements from testicular cancer patients. There were no significant differences in the frequencies of chromosomal aberrations between controls and cancer patients (9.7 and 10.3% respectively; P = 0.4921). Our results indicate that spermatozoa from untreated testicular cancer patients do not show an increased chromosomal instability as compared to control donors. PMID- 10374130 TI - Secondary infertility as early symptom in a man with multiple endocrine neoplasia type 1. AB - Multiple endocrine neoplasia-type 1 (MEN1) is an autosomal dominant familial cancer syndrome characterized by parathyroid hyperplasia, pancreatic endocrine tumours and pituitary adenomas. Here, we report a patient with a history of insulinoma who developed secondary infertility as a further symptom of the disease. When he was first examined at the age of 36 years, he complained of weakness, reduced libido and impotence. Laboratory evaluation revealed non obstructive azoospermia and hyperprolactinaemia. In contrast to sexual activity and serum prolactin, semen quality did not significantly respond to bromocriptine therapy. During follow-up, a growing pituitary adenoma caused acromegaly with elevated serum concentrations of growth hormone, insulin-like growth factor 1 (IGF-1), and prolactin. After microsurgery of the tumour at the age of 44 years, sperm concentration persistently increased up to 5.6 x 10(6)/ml. In accordance with the clinical diagnosis of MEN1, DNA sequencing revealed a mutation in exon 2 of the menin gene which results in a truncated, inactive protein product. In conclusion, MEN1 with pituitary lesions may cause severe hypogonadism and infertility. Both hyperprolactinaemia and overproduction of growth hormone and IGF-1 seem to be involved in testicular dysfunction in the present case. The possible role of menin in the testis, however, remains to be elucidated. PMID- 10374131 TI - Psychosocial experiences in women facing fertility problems--a comparative survey. AB - In a survey involving 281 patients awaiting assisted reproduction treatment at five centres in three countries, and 289 population controls, we investigated whether the patients had experienced more negative emotional feelings and negative emotional impact during periods when they were attempting to conceive as compared with the controls, and whether there was any difference in their well being at the time of consultation. The study was performed in the context of currently divergent views as to the burden of fertility problems. The survey was carried out using questionnaires of the self-administration type. Women with fertility problems did in fact consistently report a higher prevalence of negative emotions than the controls with reference to the periods during which they had been trying to conceive. Patients reported more changes in interpartner relationships (either negative or positive). Sexuality was negatively affected among the patients. At the time of consultation, the patients had less favourable scores than the controls on scales for depressed mood, memory/concentration, anxiety and fears, as well as for self-perceived attractiveness. One in four (24.9%) of the patients had scores indicating depressive disorders as compared with only 6.8% of the controls. Current well-being was even more markedly affected in patients with previous unsuccessful in-vitro fertilization (IVF) experience. The 'infertility' life event was perceived as severe by both patients and controls. Both prior to consultation and during diagnosis and treatment, women with fertility problems had a higher prevalence of reported negative psycho emotional experiences than women without fertility problems. PMID- 10374132 TI - Organizational selection and assessment of women entering a surrogacy agreement in the UK. AB - In the UK, surrogacy procedures are unregulated and not monitored. Information concerning the selection and assessment of intended (the mother commissioning) and surrogate mothers (the mother carrying and delivering the baby) is therefore not generally available (BMA, 1996). It is important to determine what type of assessment is used, and how selection takes place within the organizations dealing with surrogate motherhood arrangements. The present survey enquired about the incidence, selection and assessment procedures of all registered surrogate and commissioning couples, and aimed to find out what advice and support is given. Eight organizations took part in the survey, six were clinics and two agencies dealing with surrogate arrangements. Two voluntary organizations/helplines were also surveyed, but their data are not relevant to the results presented here. An interview and questionnaire approach was used. Psychosocial assessment was minimally addressed by all organizations, and no fixed procedures for assessment and selection were employed. Despite this, few incidences of controversial cases were reported. Confidence in this practice could be increased in the future if both parties embarking on a surrogacy arrangement knew they were properly selected and assessed. A regulatory body could monitor consistent use of professional evidence-based criteria prior to arrangements. PMID- 10374133 TI - Difficulties in distinguishing between a mature spermatid and testicular spermatozoon. PMID- 10374134 TI - Chlamydia antibody titres. PMID- 10374135 TI - Laparoscopic findings after ultrasound-guided transvaginal ethanol sclerotherapy for ovarian endometrial cyst. PMID- 10374136 TI - Growth hormone kinetics in polycystic ovary syndrome. PMID- 10374137 TI - Pathophysiology of unilateral pleural effusions in the ovarian hyperstimulation syndrome. PMID- 10374138 TI - Depression, disease severity, and sickle cell disease. AB - The present study examined depressive symptomatology in 440 adults with sickle cell disease (SCD). Participants completed the Center for Epidemiologic Studies- Depression scale (CES-D) as part of their yearly routine visits to the Duke University--University of North Carolina Comprehensive Sickle Cell Center. They also completed questions regarding demographics, disease severity, pain, and health care use. Data analyses revealed that the percentage of patients with SCD exhibiting significant depressive symptomatology dropped from 43 to 18% when a more stringent cutoff was used on the CES-D, suggesting that future studies should determine the most valid cutoff score for identifying depression in patients with SCD. Gender and family income were positively and significantly associated with depressive symptomatology. Also, patients who reported more frequent painful episodes were more likely to report depressive symptoms. Implications for assessment and treatment of depression in adults with SCD are discussed. PMID- 10374139 TI - Assessing depressive symptoms in multiple sclerosis: is it necessary to omit items from the original Beck Depression Inventory? AB - Overlap between depression scale item content and medical symptoms may exaggerate depression estimates for patients with multiple sclerosis (MS). We reconsider Mohr and co-workers' (1997) recommendation to omit Beck Depression Inventory (BDI) items assessing work ability (item 15), fatigue (17), and health concerns (20) for MS patients. Subjects were medical patients with either MS (n = 105) or a medical disorder for which the BDI is empirically supported [diabetes mellitus (DM), n = 71; chronic pain (CP), n = 80], psychiatric patients with depressive disorder (MDD; n = 37), and healthy controls (HC; n = 80). Relative scores for the eight "somatic" BDI items were analyzed by multivariate analysis of variance with demographic variables and BDI total as covariates. The only significant difference was MS > HC (item 15). On raw scores, MS patients exceeded HCs on items 15 and 21 (sexual disinterest), but this was attributable to the low HC item endorsement. There were no other differences on somatic items or item-total correlations. Scale consistency was good across groups, regardless of item omission. Somatic items were unassociated with major MS parameters. We thus encourage continued application of the full BDI for assessing depressive symptoms in patients with MS. PMID- 10374140 TI - Chronic minor stressors and major life events experienced by low-income patients attending primary care clinics: a longitudinal examination. AB - Chronic minor stressors and major life events were assessed from 129 randomly selected low-income patients attending primary care medical clinics. Participants reported experiencing an average of 15 chronic minor stressors in a 12-month period. The most common chronic minor stressors were reported in the areas of finances and domestic activities. Participants also reported these stressors as the most intense. The t tests revealed that female participants reported significantly (p = .05) more chronic events than males. The most common major life events were reported in the areas of vegetative symptoms (i.e., major change in sleeping and eating habits), financial status, illness, and interpersonal relationships. The most stressful life events were changes in vegetative symptoms, family illness, and interpersonal relationships. The t tests revealed that employed participants reported significantly (p < .05) more positive life events than did unemployed participants. Implications of the findings are discussed. PMID- 10374141 TI - Psychosocial adjustment and mental health two months after coronary artery bypass surgery: a multisystemic analysis of patients' resources. AB - Resources related to cardiac patients' sense of self, marital quality, and social support were assessed before their first planned bypass surgery to determine their relationship with later psychosocial functioning. Six female and 45 male cardiac patients, 45-70 years old, answered self-report instruments 1 week before and 8-10 weeks after the operation. Regression analyses indicated that only the marital relationship variables made independent contributions to the prediction of functioning. Since self-report measures of resources can be affected by negative affectivity, a second level of analysis predicted change scores while controlling for initial levels of functioning. Marital flexibility and support were found to make an independent contribution to recovery. These results highlight the importance of marital resources in coping with the acute phase following bypass surgery and have implications for prevention and clinical practice. PMID- 10374142 TI - Daily psychosocial factors predict levels and diurnal cycles of asthma symptomatology and peak flow. AB - This study examines the relationship among psychosocial factors, asthma symptoms, and peak expiratory flow rate (PEFR) in the natural environment. Twenty adult asthmatics wore preprogrammed wristwatches that prompted them to assess PEFR, asthma symptoms, and psychosocial factors five times a day for 10 days. Psychosocial variables (activities, locations, social contacts, mood, and stressors) were strongly related to PEFR and asthma symptoms, suggesting that they may play a more important role in disease expression than has been previously thought. Diurnal cycles of asthma symptoms and PEFR were observed. However, statistically controlling for psychosocial factors eliminated diurnal cycles for PEFR or asthma symptoms, indicating that psychosocial factors are a major contributor to the observed diurnal cycle in PEFR and symptoms. These relationships underscore the need to include psychosocial factors in future asthma research. PMID- 10374143 TI - Acculturation and cigarette smoking among African Americans: replication and implications for prevention and cessation programs. AB - We report a replication of a 1996 study on the role of acculturation in smoking among African American adults. Results with the current sample were nearly identical to the prior ones: smokers tended to be traditional and nonsmokers acculturated, with nearly 70% of Black smokers in both studies being highly traditional in their cultural orientation. Given that coming from a highly traditional Black family was a strong predictor of smoking in both studies, we suggest that new smoking prevention and cessation programs might be culturally tailored for Blacks by focusing on smoking as a familywide issue. PMID- 10374144 TI - Face to face with IOLs. PMID- 10374145 TI - Reporting innovation. PMID- 10374146 TI - Harold Ridley's first patient. PMID- 10374147 TI - Refractive changes after IOL implantation. PMID- 10374148 TI - Double phaco chop. PMID- 10374149 TI - Patient survey to determine the necessity of Internet exposure for a general ophthalmology practice. PMID- 10374151 TI - Small peripheral anterior continuous curvilinear capsulorhexis. AB - Cataract surgery is routinely performed using an anterior continuous curvilinear capsulorhexis (CCC). A manual surgical technique is described for performing a small (less than 1.5 mm diameter) anterior CCC. This technique's applications extend from Phaco-Ersatz, a cataract surgical technique designed to restore accommodation to pediatric cataract surgery. An experimental rabbit study was conducted to determine the feasibility of the technique. Up to 9 small peripheral anterior CCCs were made in the same lens capsule without the capsule tearing. The mean diameter of the CCCs was 1.1 mm +/- 0.3 (SD). A 30 gauge needle and Utrata capsulorhexis forceps were used to construct the CCC. This technique shows promise for the successful performance of small CCCs in Phaco-Ersatz procedures and pediatric cataract surgery. PMID- 10374150 TI - Consultation section. Cataract surgical problem. PMID- 10374152 TI - Analysis of edge glare phenomena in intraocular lens edge designs. AB - PURPOSE: To determine the image and relative intensity of reflected glare images from 4 commonly used intraocular lens (IOL) edge designs to assess the potential for noticeable postoperative edge glare. SETTING: University of Texas Medical School, Houston, Texas, USA. METHODS: The interaction of light rays from 4 common IOL edge designs were examined in an eye model using the OptiCAD 3-D radiometric ray-tracing program (Opticad Corp.). Comparison of the potential of the 4 edge designs to produce visual sensations was derived from plots of the spatial location and energy distribution of rays forming the retinal image. RESULTS: Edge designs with no anterior and posterior dioptric powers at the lens periphery (lenticular) and rounded corners distributed the edge glare rays over a large retinal area. Edge designs with sharp edges formed by "cropping" the anterior and posterior optic zones focused edge glare rays into distinct arc-shaped images. The peak intensity of the arc-shaped image was 8 to 10 times stronger than the peak intensity of the diffuse image formed by lenses with rounded edges. CONCLUSIONS: Rounded IOL edges distribute reflected glare image over a significantly greater area than sharp edges. Rounded edges reduce the potential for edge glare phenomena that appear to the patient as a thin crescent or partial ring. PMID- 10374153 TI - Simultaneous bilateral cataract extraction. AB - PURPOSE: To evaluate the visual outcome and safety of simultaneous bilateral cataract extraction. SETTING: Stobhill Hospital NHS Trust, Glasgow, United Kingdom. METHODS: This retrospective case review comprised 259 consecutive patients (518 eyes) who had simultaneous bilateral cataract surgery. Surgeries included bilateral extracapsular procedures, uniocular extracapsular procedures performed simultaneously with a different type of intraocular lens surgery in the other eye, and 1 bilateral intracapsular procedure. Outcome measures were postoperative best spectacle-corrected visual acuity (BSCVA), intraoperative and postoperative complication rates, and conjunctival swab culture results. RESULTS: Eighty-three percent of patients (75% of eyes) with measured preoperative and postoperative BSCVA achieved an acuity of 6/12 or better. Intraoperative and postoperative complication rates were similar to those in previous reports of unilateral extracapsular surgery and simultaneous bilateral cataract surgery. Endophthalmitis occurred in 1 eye (0.19%). There were no bilateral complications that resulted in visual loss. Cultures were positive from 42% of conjunctival swabs; 81% of positive cultures were coagulase-negative Staphylococcus and 10% were Staphylococcus aureus. CONCLUSIONS: Simultaneous bilateral cataract surgery did not lead to an increased incidence of serious intraoperative or postoperative complications, and visual acuity results were good. PMID- 10374154 TI - Primary posterior capsulorhexis without anterior vitrectomy in pediatric cataract surgery: longer-term outcome. AB - PURPOSE: To assess the effectiveness of primary posterior capsulorhexis without anterior vitrectomy in preventing posterior capsule opacification (PCO) in pediatric cataract surgery. SETTING: Children's Hospital, Dublin, Republic of Ireland. METHODS: The study comprised 32 eyes of 22 pediatric patients who had cataract extraction between 1994 and 1998. Extracapsular cataract extraction was performed using radiofrequency diathermy capsulorhexis to the anterior and posterior capsules without an anterior vitrectomy. Posterior chamber intraocular lens implantation was performed in 20 eyes. There were 23 congenital, 6 developmental, and 3 traumatic cataracts. RESULTS: Patient age ranged from 1 month to 12 years. Mean follow-up was 19 months (range 6 to 50 months). Twenty seven of 32 eyes (84.4%) had a clear visual axis at last follow-up. Five eyes required a neodymium: YAG capsulotomy, which was performed a mean of 5 months postoperatively (range 1 to 9 months). The incidence of PCO requiring capsulotomy was 15.6%. CONCLUSION: Primary posterior capsulorhexis without anterior vitrectomy was safe and effective, with a low reopacification rate. Long-term follow-up of this patient cohort is necessary. PMID- 10374155 TI - Primary capsulectomy, anterior vitrectomy, lensectomy, and posterior chamber lens implantation in children: limbal versus pars plana. AB - PURPOSE: To compare the results of a limbal versus a pars plana approach for primary posterior capsulectomy and anterior vitrectomy in the management of childhood cataract. SETTING: Department of Ophthalmology, Labbafinejad Medical Center, Tehran, Iran. METHODS: A randomized, controlled, double-masked clinical trial of 45 eyes was conducted. After being matched, 38 eyes were included in the study and were divided into 2 equal groups for data analysis. All eyes had lensectomy and posterior chamber intraocular lens (PC IOL) implantation. Primary posterior capsulectomy and anterior vitrectomy were performed through the limbus in half of the eyes and the pars plana in the other half. Main outcome measures included visual acuity, estimated red reflex, postsurgical inflammatory reaction, corneal clarity, posterior synechias, iris capture, IOL position, capsulectomy size, glaucoma, cystoid macular edema, retinal tear, and postoperative refraction. RESULTS: No statistically significant differences were found between the 2 approaches in the outcome measures. CONCLUSION: The anatomic and visual results were encouraging when posterior capsulectomy and anterior vitrectomy, using a limbal or pars plana approach, were combined with lensectomy and PC IOL implantation in children. The application of these techniques depends on surgeon experience and skill. PMID- 10374156 TI - Sulcus fixation without capsular support in children. AB - PURPOSE: To evaluate long-term follow-up in eyes of children who had sulcus fixation of an intraocular lens (IOL) without capsular support. SETTING: St. Eriks Eye Hospital/Karolinska Institute, Stockholm, Sweden. METHODS: This retrospective study included 21 eyes of 13 children. Seven eyes had Marfan's syndrome, 7 essential lens dislocation, 2 perforation with lens injury, and 5 spherophakia. The IOL implantation was primary in 16 eyes and secondary in 5 eyes. Lensectomy was performed with a limbal approach. An IOL with holes in the haptics was sutured in the sulcus, with the knots buried in the scleral bed. RESULTS: Mean patient age was 5.8 years +/- 2.6 (SD). Follow-up ranged from 9 to 33 months. No complications occurred during surgery. In all cases after IOL implantation, best corrected visual acuity was equal to or better than preoperatively. After surgery, no opacification of the visual axis, secondary glaucoma, or retinal complication was recorded. Posterior synechia formation occurred in 4 eyes, and 4 had cells on the IOL surface in 2 eyes, the IOL optic subluxated into the anterior chamber with the haptics in place. Both cases were successfully treated with pilocarpine 4%. CONCLUSION: Our results suggest that sulcus fixation of an IOL without capsular support is an option to correct aphakia in children. PMID- 10374158 TI - Influence of phacoemulsification and intraocular lens implantation on the course of diabetic retinopathy. AB - PURPOSE: To study the effect of phacoemulsification and posterior chamber intraocular lens implantation on the course of diabetic retinopathy using the nonoperated fellow eye as a control. SETTING: Departments of Ophthalmology, Diabetes Center, Tokyo Women's Medical University, and University of Tokyo School of Medicine, Tokyo, Japan. METHODS: One eye of 66 diabetic patients who preoperatively had a similar stage of retinopathy in both eyes or no retinopathy bilaterally had cataract surgery. The course of diabetic retinopathy was followed for 1 year postoperatively. Patients were placed into 1 of 2 groups: Group A, progression of retinopathy in the operated eye was attributable to the surgical invasion (i.e., there was progression of retinopathy only in the operated eye or more progression in the operated eye than in the nonoperated fellow eye); Group B, no deterioration of retinopathy bilaterally, comparable level of deterioration in both eyes, or greater progression in the nonoperated eye than in the operated eye. RESULTS: Surgery resulted in retinopathy progression in 16 patients (24.2%, Group A): 13 with unilateral deterioration and 3 with greater progression in the operated than in the nonoperated fellow eye. Of the remaining 50 patients (75.8%, Group B), 39 presented no significant progression in either eye, 8 had bilaterally comparable progression, and 3 showed progression in the nonoperated fellow eye only. Retinopathy worsened in the operated eye in 24 cases (36.3%); of these, changes in 16 patients were attributed to surgical influence. There was no significant difference between Groups A and B in age, diabetes mellitus duration, diabetes treatment method, and preoperative and postoperative hemoglobin A1c levels. The distribution of preoperative retinopathy stage significantly differed between groups, with more patients without retinopathy in Group A and more patients with advanced retinopathy in Group B. CONCLUSION: Factors such as age, diabetes mellitus duration, diabetes treatment method, and hemoglobin A1c level did not affect the progression of retinopathy; however, preoperative status of retinopathy may influence the susceptibility of the retinopathy to surgical invasion. A considerable proportion of eyes with aggravation of retinopathy would reflect the natural course of the disease, systemic factors, or both rather than the influence of cataract surgery. PMID- 10374157 TI - Heparin-surface-modified intraocular lenses in pediatric cataract surgery: prospective randomized study. AB - PURPOSE: To evaluate the performance of heparin-surface-modified (HSM) intraocular lenses (IOLs) in pediatric eyes after cataract surgery. SETTING: L.V. Prasad Eye Institute, Hyderabad, India. METHODS: This prospective, randomized, double-masked, controlled clinical trial comprised 90 children aged 2 to 14 years with cataract. The patients were consecutively randomized to receive an HSM (Group 1) or an unmodified (Group 2) poly(methyl methacrylate) (PMMA) IOL. Extracapsular cataract extraction (ECCE) with IOL implantation was performed in children 8 years and older and ECCE with primary posterior capsulotomy, anterior vitrectomy, and IOL implantation in children younger than 8 years. Outcome parameters were inflammatory cell deposits on the IOL surface, posterior synechias, and anterior chamber reaction. RESULTS: Follow-up data were available for 73, 70, 60, and 68 patients at 1 week, 1 month, 3 months, and 6 months, respectively. Significantly fewer cell deposits were noted in Group 1 at 1, 3, and 6 months (P < .001). Synechia formation and anterior chamber reaction were comparable in the 2 groups. CONCLUSION: The lower incidence of inflammatory cell deposit formation in eyes with HSM PMMA IOLs indicates that these IOLs have greater bicompatibility than unmodified IOLs in pediatric cataract surgery. PMID- 10374159 TI - Effects of NG-nitro L-arginine and corticosteroids on aqueous humor levels of nitric oxide and cytokines after cataract surgery. AB - PURPOSE: To assess the efficacy of nitric oxide synthesis (NOS) inhibitor, topical steroids, and nonsteroidal anti-inflammatory drugs on aqueous levels of nitric oxide (NO) and cytokines after cataract surgery. SETTING: Research Laboratory, Inonu University Turgut Ozal Medical Center, Malatya, Turkey. METHODS: Fifteen rabbits had intercapsular phacoemulsification and were randomly divided into 3 treatment groups: Group 1 was treated with topical prednisolone acetate 1% drops 5 times a day for 1 week; Group 2, flurbiprofen 0.03% drops 5 times a day for 1 week; Group 3, a 0.1 cc subconjunctival injection of NG-nitro L arginine (L-NAME) (150 mg/kg) 1 day and 3 days after surgery. Three rabbits serving as controls received a subconjunctival injection of an equal volume of balanced salt solution (BSS) at the same times as the L-NAME injections. Aqueous humor specimens were collected from each eye to determine NO and cytokine levels including interleukin-1-beta (IL-1 beta), interleukin-2R (IL-2R), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha). RESULTS: The levels of IL-1 beta and IL-6 were higher in Group 2 and the control group than in Groups 1 and 3 at all times. The differences were not statistically significant (P < .05). Nitric oxide and TNF-alpha levels in Groups 1 and 3 were significantly lower than in Groups 2 and the controls 1, 3, and 7 days postoperatively (P < .05). CONCLUSION: These findings suggest a strong inhibitory effect of NOS inhibitors and corticosteroids on aqueous levels of TNF-alpha and NO and no inhibitory effect on IL-1 beta and IL-6 levels after cataract surgery. PMID- 10374160 TI - Dodick laser phacolysis: thermal effects. AB - PURPOSE: To gather experimental data on whether Dodick laser phacolysis leads to corneal or scleral burns. SETTING: The Eye Department, County Hospital Salzburg, Salzburg, Austria. METHODS: The study was done using a pulsed neodymium:YAG (Nd:YAG) laser with a wavelength of 1064 nm; energy, 10 mJ; and duration of pulses, 14 ns. The light pulse is carried by a 400 microns quartz fiber to the laser phacolysis probe. The laser light hits a titanium target inside the tip, causing an optical breakdown and thus a shock wave. The generation of both plasma and the shock disrupt the nuclear material. The temperature at the ultrasonic phaco and laser phacolysis tip was measured under air and balanced salt solution (BSS) in a test chamber and in the anterior chambers of eye-bank eyes. RESULTS: Ultrasonic phacoemulsification led to a difference in temperature up to 55.3 degrees C under air, 12 degrees C in BSS, and 10.9 degrees C in the anterior chamber. There was no clinical significant heat generated by the laser phacolysis tip. CONCLUSION: This initial in vitro study demonstrates that the well-known risk of the tissue heating (i.e., phaco burn) does not occur with Dodick laser phacolysis, even when the irrigation flow is slow or discontinued. PMID- 10374161 TI - Effect of astigmatism on multifocal intraocular lenses. AB - PURPOSE: To study the effect of astigmatism on multifocal intraocular lens (MIOL) function. SETTING: Laser Laboratory, Department of Electronics, Electrotechnics and Informatics, University of Trieste, Italy. METHODS: Using an experimental optical system, this study compared the division of a laser beam in the focal spots of 1 monofocal IOL (Pharmacia 722A), 1 bifocal IOL (Pharmacia 811E), and 2 MIOLs (AMO Array MPC25NB and Domilens Progress 1). The model consists of a helium neon laser and an optical system: a triangular optical bench with a precision collimator, a micropositionable immersion stage to support the IOL, and a digital image-processing system. Astigmatism was induced by interposing a +1.0 diopter (D) cylinder lens between the IOL and the television camera. Astigmatism was corrected by adding a -1.0 D cylinder lens in front of the IOL on the same axis. RESULTS: Astigmatism creates pairs of focal lines, 1 for each focal spot in the IOL. In the multifocal IOL, the posterior focal line of the nearest focus interfered with the anterior line of the next focus. Correcting the astigmatism led to a significant reduction (mean 20%) in light intensity. CONCLUSIONS: Astigmatically neutral surgery or surgical correction of pre-existing astigmatism is essential in MIOL implantation to minimize the decrease in contrast sensitivity. PMID- 10374162 TI - Implantation of a black diaphragm intraocular lens for traumatic aniridia. AB - PURPOSE: To evaluate the suitability and safety of a black diaphragm posterior chamber intraocular lens (IOL). SETTING: Department of Ophthalmology, Leeds General Infirmary, Leeds, United Kingdom. METHODS: Seven patients who had secondary implantation of a Morcher 67G black diaphragm posterior chamber IOL were identified. All patients were men with a mean age of 42 years who had previous ocular trauma resulting in extensive loss of iris tissue (traumatic aniridia). Simultaneous penetrating keratoplasty was performed in 4 cases. Minimum follow-up was 10 months (mean 19 months). RESULTS: Best corrected visual acuity improved in 5 cases and was unchanged in 1 case. The lens was well centered in 5 cases. Two cases developed secondary glaucoma, 1 requiring trabeculectomy. One case developed infective endophthalmitis but had a visual acuity of 6/18 at last follow-up, and 1 had a vitreous and anterior chamber hemorrhage, which resolved. CONCLUSIONS: The black diaphragm posterior chamber IOL overcame aphakia in eyes with considerable loss of iris tissue and may mitigate the visually disabling effects of traumatic aniridia. Although this lens appears safe, caution should be used in its implantation until more patients with longer follow-up are studied. PMID- 10374163 TI - Evaluation of relationships among refractive and topographic parameters. AB - PURPOSE: To examine the relationships among several refractive and topographic parameters. SETTING: Cullen Eye Institute Department of Ophthalmology, Baylor College of Medicine, Houston, Texas, USA. METHODS: Using computerized videokeratography (EyeSys Corneal Analysis System), 287 corneas of 150 patients were retrospectively analyzed. The Holladay Diagnostic Summary (HDS) refractive maps were used to evaluate relationships among variables of the HDS and refractive error. RESULTS: Myopic spherical equivalent refraction (P = .0003) and more negative asphericity (Q-values) (P = .0119) were correlated with steeper corneas. The Q-values were less negative in eyes with moderate myopia (2.0 to 6.0 diopters [D]) than in those with hyperopia (1.0 D or greater). The Q-values below -0.3 were correlated with less favorable values for predicted corneal acuity and corneal uniformity index values. Mean corneal curvature measurements obtained by computerized videokeratography and standard keratometry showed a strong degree of correlation (P = .0001). CONCLUSION: As the degree of myopia and negative asphericity increased, the corneal radius of curvature decreased. Corneal Q values less than -0.3 were associated with reduced optical performance of the cornea. PMID- 10374164 TI - Preliminary tests and construction of a computerized quantitative surgical keratometer. AB - PURPOSE: To construct a simple, computer-based, quantitative surgical keratometer to measure a 3.0 to 4.0 mm central region of the corneal surface. SETTING: Laboratorio de Optica Oftalmica, Instituto de Fisica de Sao Carlos (IFSC-USP), Sao Paulo, Brazil. METHODS: A high-intensity fiber-optic-illuminated ring pattern (Placido disk) is projected on the cornea. Reflected images are captured by a charge-coupled device camera mounted on a Zeiss microscope beam splitter and then digitized by a frame grabber installed on an IBM-compatible personal computer. Simple algorithms based on image-processing techniques were implemented for border detection. A calibrating curve based on 4 spherical surfaces was used to calculate diopter values at 360 points at each examination. Results were plotted on the computer monitor using diopter value versus angle (1 through 360 degrees) graphs. Preliminary measurements of 14 healthy corneas were compared with the equivalent radial distance points measured on an EyeSys corneal topographer. RESULTS: Mean deviation was 0.05 mm for radius of curvature, 0.24 diopter for power, and 5 degrees for cylinder. CONCLUSION: The keratometer provided precise measurements for corneal shape control during surgery. PMID- 10374165 TI - Comparison of objective and subjective refraction before and after laser in situ keratomileusis. AB - PURPOSE: To compare the accuracy and reliability of objective and subjective refractions before and after laser in situ keratomileusis (LASIK) for myopia, hyperopia, and astigmatism. SETTING: Augenchirurgie und Laserzentrum Hochrum, Innsbruck, Austria. METHODS: In this prospective study, the objective refraction obtained with the Nidek AR-K 900 autorefractor was compared with the subjective refraction in 159 eyes (125 with myopia and 34 with hyperopia) operated on with 2 different lasers. Refractions were done before and 6 months after LASIK. RESULTS: Preoperatively, the objective and subjective refractions correlated better in eyes with low myopia than in those with high myopia (P < .01). Postoperatively, objective refraction was less accurate and reliable than preoperatively. The difference between the objective and subjective spherical refractions was statistically significant (P < .0001) after LASIK in eyes with hyperopia. The correlation between the objective and subjective cylindrical refractions was stronger preoperatively. Especially after LASIK for hyperopia, the objective refraction did not reliably assess the magnitude and axis of the cylinder. The preoperative refractive error did not significantly affect the preoperative and postoperative difference between the objective and subjective refractions or the change between the preoperative and postoperative mean differences. The type of excimer laser used significantly affected the difference between the objective and subjective refractions. CONCLUSIONS: Especially after LASIK for hyperopia, the objective refraction determined with the Nidek AR-K 900 autorefractor delivered erroneous results, which have implications for postoperative care and preoperative measurements for ocular surgery such as enhancement procedures or cataract surgery. PMID- 10374166 TI - Effects of topical diclofenac and prednisolone eyedrops in laser in situ keratomileusis patients. AB - PURPOSE: To compare the effects of a topical nonsteroidal anti-inflammatory agent, diclofenac, and prednisolone acetate on wound healing, postoperative inflammation, and other clinical parameters in laser in situ keratomileusis (LASIK) patients. SETTING: Fundacion VER, Cordoba, Argentina. METHODS: Laser in situ keratomileusis for myopia was performed simultaneously in both eyes of 16 patients by 1 surgeon. Patients were prospectively randomized to receive diclofenac eyedrops in the right eyes and prednisolone acetate eyedrops in the left. Postoperatively, the drops were administered topically every 2 hours the first day, every 6 hours the first week, every 8 hours the second week, and once a day the fourth week. Preoperatively and at each postoperative visit, evaluation of visual acuity, slitlamp biomicroscopy, corneal topography, and clinical scoring (0-III) of pain and photophobia were done; epithelial interface opacities were objectively evaluated. Follow-up was at 24, 48, and 72 hours, 1, 2, and 3 weeks, and 1, 2, 3, and 6 months. A paired t test was used for statistical analysis. RESULTS: Mean age of the 16 patients was 29.4 years +/- 6.5 (SD). Preoperatively, mean spherical equivalent was -5.83 +/- 3.61 diopters (D) in the right eyes and -6.96 +/- 4.66 D in the left eyes. At 6 months postoperatively, it was -1.83 +/- 1.87 D and -1.88 +/- 2.13 D, respectively. In the first 24 hours, there were significant clinical symptoms in the diclofenac group. CONCLUSIONS: Wound healing was stable, with no regression, in the diclofenac and prednisolone groups. Both anti-inflammatory agents worked well in LASIK patients. PMID- 10374167 TI - Comparison of wound healing after photorefractive keratectomy and laser in situ keratomileusis in rabbits. AB - PURPOSE: To evaluate and compare the corneal wound-healing process after photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK). SETTING: Kangnam St. Mary's Hospital, Seoul, Korea. METHODS: Two surgical procedures, PRK with the VISX Star excimer laser and LASIK with a MicroTech microkeratome, were performed in 24 rabbit eyes. In the PRK group (n = 12 eyes), the rabbit cornea was treated with a 20 microns ablation. In the LASIK group (n = 12 eyes), a 20 microns laser ablation was performed after a 150 microns thick hinged corneal flap had been made. During both procedures, dichlorotriazinyl aminofluorescien (DTAF) dye was applied to the ablated stromal bed; in the LASIK group, the stromal side of the corneal flap was also stained with DTAF to differentiate regenerated collagen from normal stromal tissue. Corneal wound healing was evaluated postoperatively at 1, 4, 8, and 12 weeks using light, electron, and fluorescence microscopy. The amount of regenerated stromal tissue and the number of keratocytes were analyzed by an image-analysis system. RESULTS: In the PRK group, epithelial migration and regeneration were observed in the ablated area without any stromal regeneration 1 week postoperatively. However, newly regenerated, irregularly arranged stromal collagen, with epithelial hyperplasia in the ablated area, was observed 4 to 12 weeks postoperatively by light and fluorescence microscopy. The number of keratocytes in the surgical area was also increased. In ultrastructural observation using an electron microscope, the shape of keratocytes in the ablated area was changed, and the number of rough and smooth endoplasmic reticuli, ribosomes, mitochondria, and electron-dense vesicles in the cytoplasm were increased, suggesting that the cells were activated. In the LASIK group, there was no observed regenerated collagen between the corneal flap and the ablated stromal bed except in the wound margin. Lamellated, parallel collagen fibers in the cornealstroma were not disturbed. However, in the wound margin, corneal epithelial ingrowth between the flap and the stromal bed was observed, as was some regenerated stromal tissue. The amount of regenerated stromal tissue and the number of keratocytes in the wound area were statistically smaller than those in the PRK group (P < .05). Observation by electron microscopy showed no activated keratocytes, unlike in the PRK group. The collagen fibers in the wound area were parallel. CONCLUSION: Stromal wound healing in the LASIK group was minimal compared with that in the PRK group, except in the wound margin. These results may support the clinical findings of less corneal haze in the human cornea after LASIK. PMID- 10374168 TI - Practice styles and preferences of ASCRS members--1998 survey. AB - A survey of the practice styles and preferences of members of the American Society of Cataract and Refractive Surgery with United States ZIP codes was performed in September 1998. Approximately 29% (1434) of the 5000 questionnaires mailed were returned before the November cutoff date. Three profile questions were used to cross-tabulate: age of the respondent, geographic location, and volume of cataract surgery per month. Current data were compared with data in previous annual surveys. PMID- 10374169 TI - Contact zone of piggyback acrylic intraocular lenses. AB - In a hyperopic cataract patient, surgery was performed with implantation of 2 foldable, acrylic, posterior chamber intraocular lenses (IOLs) in the bag. The IOLs showed a central contact zone during surgery. This contact zone remained after surgery and was documented 2 months postoperatively. The contact zone may induce multifocality similar to that seen with multifocal IOLs. PMID- 10374170 TI - Interpseudophakos Elschnig pearls associated with late hyperopic shift: a complication of piggyback posterior chamber intraocular lens implantation. AB - We report 3 cases of bilateral piggyback lens implantation in which late hyperopic shift occurred associated with Elschnig pearl formation in the peripheral interface between the 2 lenses. PMID- 10374171 TI - Pseudo-Fleischer ring after hyperopic laser in situ keratomileusis. AB - A 37-year-old woman had bilateral hyperopic laser in situ keratomileusis (LASIK). Six months postoperatively, an epithelial pigmentation ring pattern was identified on both corneas. The appearance of the ring pattern was similar to the iron deposits of the Fleischer ring of keratoconus. While corneal iron deposits in various patterns have been reported after other forms of ocular therapeutic and refractive surgery, this is the first report of the association between hyperopic LASIK and a corneal iron line, which we have called the "pseudo Fleischer ring." PMID- 10374172 TI - Corneal topography of spontaneous perforation of acute hydrops in keratoconus. AB - An unusual case of spontaneous corneal perforation of acute hydrops in the left eye of a 21-year-old man is presented. The patient had a history of atopic diseases. To evaluate the status of the other eye, corneal topographic analysis was performed. This confirmed a subclinical keratoconus in the fellow eye and the association with atopy, eye rubbing, and rapid progression of the ectasia leading to perforation. PMID- 10374173 TI - Severe iridodialysis from phacoemulsification tip suction. AB - During cataract surgery, the iris of an 83-year-old woman was strongly sucked into the phacoemulsification tip twice, resulting in severe iridodialysis. The dehisced iris was sutured to the sclera using double-armed 10-0 polypropylene on a long curved needle. Intensive suction of the iris by the phacoemulsification tip can lead to severe iridodialysis. Machine setting parameters, particularly flow rate, should be lowered after the first iris suction to avoid further iris damage. PMID- 10374174 TI - [The effectiveness of recombinant growth hormone in height deficiency due to intrauterine growth retardation]. AB - BACKGROUND: Intrauterine growth retardation (IUGR) is considered to be responsible for approximately 20% of short stature in adulthood. Although GH secretion is normal in the majority of cases, excellent results have been published by some authors using GH to treat children with height deficiency due to IUGR. PATIENTS AND METHODS: Thirty children with a history of IUGR with chronological ages between 2 and 7 years and height less than 2 SD were randomized in two groups for one year: a) control group, no treatment, 14 cases, and b) treatment group, 1 U/kg/week of recombinant GH, 16 cases. Growth and maturation were analysed periodically in both groups. In addition, serum levels of GH, IGF-I, IGFBP3 and GHBP were measured before and under treatment and adverse events were assessed in treatment group. RESULTS: In the treated group significant increments in growth rate, cm/year (median = 6.91 vs 9.94), improvement in height SDS (median = -2.19 vs -1.63) and positivation of growth rate (median = -0.13 vs 3.17) were observed compared with the control group. Bone age evolved parallelly to chronological age and the height age/bone age ratio increased throughout the study under GH therapy. Hormonal findings in the treated group showed a significant increase in IGF-I and IGFBP3 values. Glycaemia levels increased without exceeding upper normal levels in the treated group. CONCLUSION: GH was effective in promoting growth in this short-term study in children with height deficiency due to IUGR. Close follow-up is required to detect any adverse event, particularly those related to carbohydrate metabolism. PMID- 10374175 TI - [Molecular study of red cell pyruvate kinase deficiency in 15 patients with chronic hemolytic anemia. Group of Erythropathology of Hematology and hemotherapy Association of Spain (HHAS)]. AB - BACKGROUND: Identification of RBC pyruvate-kinase (PK) gene mutations by polymerase chain reaction (PCR) and single strand conformation polymorphism (SSCP) followed by PK gene sequencing in positive cases has been assessed and the results obtained with a preliminary study of 15 unrelated patients of Spanish origin are presented. PATIENTS AND METHODS: Patients have been classified into two different groups: group 1, propositus (15 cases), and group 2, relatives of the patients included in group 1 (10 males and 5 females). In group 1, a PCR was followed by SSCP and sequencing, and in group 2, the PCR was followed by digestion with specific restriction endonucleases (PCR-ER). RESULTS: Group 1: from 15 patients included in the study 2 were identified as homozygous, 4 as heterozygous and 9 as compound heterozygous. In this group, were identify 26 affected alleles with 11 different mutations: T1456 10 alleles (38.6%), T721 3 alleles (11.6%), A1010, C514, C1015 and T1223 2 alleles (7.7%), and C1070, A1291, T1508, A1595 y T1675 one allele. Relatives from 8 out of 15 patients from group 1 showed the following pattern: homozygous (one case), heterozygous (10 cases), compound heterozygous (2 cases) and normal (2 cases). CONCLUSIONS: SSCP procedure followed by direct gene sequencing in positive cases is fast and simple enough to allow the identification of PK deficient variants, avoiding the need of biochemical characterisation of semipurified deficient enzyme, which is more cumbersome and time consuming. In addition, the PCR-ER method is a very useful tool for screening of the most frequent molecular variants, as well as, for the detection of the carrier condition of this enzymopathy (family studies). PMID- 10374176 TI - [Unscheduled revisits in medical emergency units at the hospital: incidence and related factors]. AB - BACKGROUND: To known the revisit rate in an emergency department (ED), to define the clinical and epidemiological profile of revisited patients, and to identify influencing factors for revisits. PATIENTS AND METHODS: During one year period, we included all revisited patients returning to ED before 72 h of a previous discharge from medical unit of ED. As controls we included the next patient seen after every case being discharged. We compiled clinical and epidemiological data from both groups. For revisited patients, we identify the cause of the revisit, changes in diagnosis and/or treatment, diagnosis mistakes and final destination of the patient. RESULTS: We identified 406 revisits (revisit rate: 1.42%). Multivariate analysis disclosed, as positive predictive factors for revisit, and age over 60 years (p = 0.006), male sex (p = 0.02), visit performed at level 2 (severe diseases) (p = 0.02), initial assessment by junior resident (p = 0.01), number of complementary procedures higher than 2 (p = 0.01) and gastrointestinal disease as diagnosis after the first visit (p = 0.03). On the other hand, dermatologic symptoms as initial complaint and unspecific symptoms (p = 0.01) were negative predictors for revisit. In only 16% of cases, the revisit did not imply changes in the diagnosis or treatment. Revisits were due to disease-related factors in 34% of cases, physician-related factors in 33%, patients-related factors in 10%, system-related factors in 3% and there were no relationship with the previous visit in 15% (in 5% of cases the cause was unassessable). The diagnosis error most frequently seen was "nonspecific abdominal syndrome". Seventy six percent of revisited patients were admitted. CONCLUSIONS: The revisit rate in our ED is relatively low. Patients being revisited have well-defined clinical and epidemiological profile. The majority of revisited patients require to be admitted. PMID- 10374177 TI - [Intrauterine treatment of growth retardation]. PMID- 10374178 TI - [Current status and new strategies in stem cell transplantation]. PMID- 10374179 TI - [Pathophysiology of pain in fibromyalgia syndrome: on the threshold of its understanding]. PMID- 10374180 TI - [Fever, pain in the right hypochondrium, cytolysis and cholestasis in a 70-year old woman]. PMID- 10374181 TI - [Prevalence of autoimmune polyglandular syndrome type II in idiopathic Addison disease]. PMID- 10374182 TI - [Fever secondary to enalapril administration]. PMID- 10374183 TI - [Idiopathic orbital myositis]. PMID- 10374184 TI - [The usefulness of the bone marrow examination in the etiological diagnosis of prolonged fever in patients with HIV infection]. AB - BACKGROUND: To analyze the usefulness of bone marrow (BM) study in the diagnosis of fever of unknown origin (FUO) in patients infected by the human immunodeficiency virus (HIV) in a single center during a period of 42 months. PATIENTS AND METHODS: 182 episodes of FUO in 162 patients p3th HIV infection were studied. Age, sex, risk factor for HIV infection, hemoglobin level, counts of leucocytes, neutrophils, lymphocytes, CD4 positive lymphocytes, platelets and levels of hepatic enzymes, albumin and beta 2-microglobulin were studied. BM aspirate was performed in all episodes for cytologic and microbiologic examination, and BM biopsy was done in 43. Analysis of factors related with the probability of diagnosis by BM examination was carried out. RESULTS: The median age was 33 years (range, 22-70), and 123 were males. Drug abuse was the most frequent risk factor for HIV infection (63%). One hundred thirty patients had previous AIDS diagnosis before they were evaluated for unexplained fever. A specific diagnosis was achieved in 161 episodes (88%) and the most frequent diagnoses were Mycobacterium spp. (55%) and Leishmania spp. (14%) infections. Fifty-four episodes (30%) were diagnosed by BM examination, and in 36 (20%) BM study was the only diagnostic tool. Examination of the BM aspirate yielded the diagnosis in 40 out of the 178 episodes (13%), whereas BM biopsy was a diagnostic tool in 8 (19%); in 9 additional cases (21%) granulomas were observed. Microbiologic study of BM smears for mycobacterial infections was positive in 28 of the 143 episodes (19%), and the culture for Leishmania was positive in 2 out of the 42 cases. The presence of thrombocytopenia (< 75 x 10(9)/l) and elevated serum levels of aspartate-aminotransferase (AST) (> 100 U/l) were the factors associated with a high probability to obtain the diagnosis through BM study. CONCLUSIONS: In patients infected by the HIV and unexplained fever, BM examination is an useful procedure for the diagnosis, particularly in areas where infections by Mycobacterium spp. and Leishmania are prevalent. So that, in our setting, systematic use of this procedure is justified for diagnosis of FUO in those patients. PMID- 10374185 TI - [Genetic analysis of RET mutations in families with multiple endocrine neoplasia type II in the community of Murcia]. AB - BACKGROUND: Multiple endocrine neoplasia type 2 (MEN 2) syndromes are inherited following an autosomal dominant pattern. RET protooncogen mutations have been associated with MEN 2. The identification of these mutations enables us to diagnose MEN 2. The objectives were to recognize RET mutations and gene carriers in the area of Murcia and to sep up the relationship between genotype and phenotype. PATIENTS AND METHODS: 284 subjects from 14 MEN 2A kindreds and one MEN 2B family from the Community of Murcia, Spain, were studied. 48 out of them had MEN 2 tumours and 236 subjects were at risk. The initial screening test was single-strand conformation polymorphism (SSCP) in 8 MEN 2A families and denaturing gradient gel electrophoresis (DGGE) in 6 MEN 2A families; the results in all the subjects were confirmed with restriction analysis. The MEN 2A family in which the Cfo-I enzyme detected but did not specify the type of mutation received DNA sequence assay. The MEN 2B kindred was studied with restriction analysis. RESULTS: TGC-->TAC and TGC-->CGC mutations of codon 634 were found in 13 and one MEN 2A kindreds, respectively. ATG-->ACG mutation of codon 918 was present in the MEN 2B family. Clinical diagnosis was confirmed in the 48 patients, 44 new gene carriers were detected and 192 carriers of normal alleles were ruled out. The incidence of hyperparathyroidism was highest if RET mutation was TGC-->CGC. CONCLUSIONS: Community of Murcia is one of the areas with the highest prevalence of MEN 2. The risk of hyperparathyroidism is increased if TGC- >CGC is present. PMID- 10374186 TI - [Polymorphism of the gene of vitamin D receptor and bone mineral density in postmenopausal women]. AB - BACKGROUND: Conflicting results have been reported on the association between restriction fragment length polymorphism at the vitamin D receptor (VDR) gene locus and bone mineral density (BMD). Population differences in environmental factors, such as calcium intake and calcidiol levels which have strong influence in BMD, may alter this association. PATIENTS AND METHODS: We analyzed the Bsml RFLP at the eight introm of the VDR gene in a population sample (n = 204) of postmenopausal Spanish women aged 50-65 years being seen clinically and studied calcium intake (dietetic questionnaire) and biochemical parameters (PTH and calcidiol). In parallel bone densitometry were measured in lumbar spine and proximal femur. RESULTS: We identified low BMD of the proximal femur in the BB group. This effect was not observed at other body locations. The calcium intake was lees than 500 mg/day in 60% of the studied population as calcidiol levels were lower than 10 ng/l in 36% of it. The total group population with normal calcium intake (> 1,000 mg/day) showed higher BMD (proximal femur and spine) than the group with low calcium intake, this variation not being observed in group BB alleles. Interestingly, we observed significant differences in BMD proximal femur between genotype groups BB versus Bb + bb when calcidiol levels were < 10 ng/l. Moreover, within the BB subgroup, those subjects with normal calcidiol levels have higher proximal femur BMD compared with those with low calcidiol levels. CONCLUSIONS: Our results indicate an effect of the VDR genotype on BMD proximal femur which is clearly influenced by calcium intake and calcidiol serum levels. PMID- 10374187 TI - [Serum lipoproteins in patients with porphyria cutanea tarda]. AB - BACKGROUND: To know the lipid profile in patients with porphyria cutanea tarda (PCT). PATIENTS AND METHODS: Serum lipoproteins have been studied in 64 males with PCT: 42 with chronic hepatitis C and 22 seronegatives for hepatitis B and C virus. Thirteen patients without porphyria, but with chronic hepatitis C, and 13 healthy subjects were also studied. RESULTS: Patients with chronic hepatitis C, with or without PCT, had a decrease of total and VLDL cholesterol (TC and VLDLc) and apolipoprotein (apo) B in comparison with healthy controls and seronegative for hepatitis C virus patients with PCT. CONCLUSIONS: Lipid abnormalities found in PCT are not related with this disease, but with the presence of chronic hepatitis C often associated to PCT. PMID- 10374188 TI - [Doctor: how much longer will I live?]. PMID- 10374189 TI - [The laborious road to evidence]. PMID- 10374190 TI - [Advance in the study of genetic basis of obesity]. PMID- 10374192 TI - [Dyslipidemia caused by ritonavir]. PMID- 10374191 TI - [65-year-old man with progressive polyneuropathy]. PMID- 10374193 TI - [Non-experimental studies and their review by ethical committees of clinical investigation]. PMID- 10374194 TI - [Treatment with intravenous immunoglobulin in pure red cell aplasia in patient wtih systemic lupus erythematosus]. PMID- 10374195 TI - [Epidemiology of the acquired pneumonia in the community of the elderly]. PMID- 10374196 TI - [Recurrent abdominal pain and vitamin E]. PMID- 10374197 TI - [Sentinel lymph node detection with lymphoscintigraphy and intraoperative probe in malignant melanoma patients]. AB - BACKGROUND: The sentinel lymph node is the first node in a lymphatic basin to receive lymphatic drainage from a tumor site. If this node is free of tumor, then radical lymphadenectomy may be avoided. The goal of this study was to assess the usefulness of lymphoscintigraphy and intraoperative gamma probe in the sentinel node detection in patients with malignant melanoma. METHOD: We prospectively studied 40 patients with malignant melanoma (24 in I/II stages and 16 in III stage). The day before surgery a lymphoscintigraphy with 99mTc-nanocolloid was performed and the first lymph node identified was considered as sentinel node. For intra-operative mapping a hand-held gamma probe was used. RESULTS: Sentinel nodes were identified in 39/40 (97.5%) patients. In 24 patients with I/II stages 34 sentinel nodes were demonstrated (six positive and 28 negative for malignant melanoma). A total amount of 161 regional lymph nodes was harvested, all of them being negative for malignant melanoma. In 16 patients with III stage, 22 sentinel nodes were located (14 positive and eight negative for malignant melanoma). A total of 89 regional lymph nodes were excised in sentinel nodes positive patients (44 positive and 45 negative for malignant melanoma) and 36 lymph nodes in sentinel node negative, all of them negative for malignant melanoma. CONCLUSIONS: In patients with malignant melanoma, lymphoscintigraphy with 99Tc-nanocolloid is useful for the detection of sentinel lymph node. Biopsy of this node is useful for the selection of patients to undertake a lymphadenectomy. PMID- 10374198 TI - [Efficacy and safety of thrombolytic therapy in pulmonary embolism: meta-analysis of randomized controlled trials]. AB - BACKGROUND: The role of thrombolytic agents in the treatment of pulmonary thromboembolism (PTE) remains a controversial issue. The objective of this study is to assess the efficacy and safety of thrombolytic therapy in the treatment of PTE by means of a meta-analysis of randomized controlled trials (RCT). METHODS: A bibliographic search of the main biomedical bibliographic databases was carried out and eight randomized controlled trials that fulfilled the inclusion criteria were found. Two blinded and independent evaluators assessed the quality of RCT according to Jadad scale, and selected the necessary data to fulfill the objective of this study. RESULTS: The selected trials were heterogeneous regarding the type of thrombolytic agent, the administration schedule, and the efficacy measures used. The methodological quality was 2 points in the Jadad scale as an average. No statistically significant differences in mortality nor in risk of PTE relapse were found between the group of patients receiving thrombolytic agents and the group not receiving them. Significant differences were found, however, between these two groups as regards the risk of bleeding events (OR = 2.62; CI 95%: 1.56-4.38). CONCLUSION: The results of these meta analyses do not suggest the use of thrombolytic therapy in PTE in everyday clinical practice since measurable risks overcome potential benefits. PMID- 10374199 TI - [Study of mortality in a medical unit of emergency department: incidence, causes and consequences]. AB - OBJECTIVES: To define the mortality pattern in a medical unit of emergency department (ED) and to know the satisfaction of relatives with ED provided care. PATIENTS AND METHODS: We computed the number of patients visited and dead from 1989 to 1996. From all patients dying during 1996, we recorded clinical and epidemiological data and we interviewed the patients' family to know their satisfaction with ED provided care. RESULTS: Whole mortality rate was 0.71 (0.15)% X (SD) with an annual increase of 10.4% (r = 0.78, p < 0.05). The clinical profile of patient dying at ED is an individual of advanced age, with a poor quality of life, and in whom the death was expected when arrived to ED. From the family interview, 61% of cases preferred that their relative was dying in the hospital, and 88% were satisfied with ED provided care. CONCLUSIONS: Although the annual mortality rate has progressively increased in ED, family satisfaction with the received care is good. PMID- 10374200 TI - [Optic neuritis, brain stem syndromes and myelitis: rapid conversion to multiple sclerosis]. AB - BACKGROUND: To establish the early risk of multiple sclerosis (MS) following isolated demyelinating syndromes. PATIENTS AND METHOD: 36 patients with optic neuritis, 24 with brainstem syndromes, 27 with spinal cord syndromes and 8 patients with a poliregional syndrome were included in a prospective protocol: including clinical data, immunological determinations, oligoclonal bands, visual, somestesic and brainstem evoked potentials and a magnetic resonance imaging (MRI) study. RESULTS: MRI showed the presence of signal abnormalities in 70% of the patients. The conversion rate (CR) was 18.5%. In patients with a normal MRI, CR was 4% compared to 26% in patients with an abnormal MRI (p = 0.05). CONCLUSIONS: The early conversion rate to MS is significantly increased when MRI is abnormal. PMID- 10374201 TI - [Intraoperative detection of sentinel lymph node in the surgery of solid tumors]. PMID- 10374202 TI - [Moral setting in medical practice]. PMID- 10374203 TI - [Study of the gene of hemochromatosis in first degree relatives of patient with porphyria cutanea tarda]. AB - BACKGROUND: Porphyria cutanea tarda (PCT) is characterized by a deficiency of uroprophyrinogen decarboxylase (URO-D). The activity of this enzyme is decreased in the presence of iron-dependent disorders. A relationship between the hepatic hemosiderosis, which is present in most patients with PCT, and the status of the hemochromatosis gene has been observed. Particularly, two mutations of the gene have been described, one of them (Cys282Tyr) is related to the iron overload and might influence on the development of the clinical signs of PCT. PATIENTS AND METHODS: We have measured the transferrin saturation percentage and observed the status of the hemochromatosis gene in a patient diagnosed with PCT and in five of their relatives. RESULTS: The proband and one sister had a marked increase of uroporphyrins and iron overload, and both had the mutation Cys282Tyr, which was also present in the mother. CONCLUSIONS: This is the first study measuring iron overload and hemochromatosis gene mutations in relatives, and not in patients with PCT. We think that the study of this family justifies the systematic investigation of these parameters in all first degree relatives of patients with PCT, to identify subjects at risk, who can benefit from prophylactic measures and early therapy. PMID- 10374204 TI - [HIV-protease inhibitors: pros and cons]. PMID- 10374205 TI - [High dose chemotherapy in germinal testicular tumors]. PMID- 10374207 TI - [Ethical dilemmas in patients with bad prognosis in intensive care units]. PMID- 10374206 TI - [Selection criteria of organ donors with respect to infectious disease transmission. Study Group on Infections in Transplantation (GESITRA). Spanish Society of Infectious Diseases and and Clinical Microbiology (SEIMS). National Organization of Transplantation (ONT). Ministry of Health Services]. PMID- 10374208 TI - [Tuberculosis in Barcelona, Spain, 1921-1998 and influence of HIV infection]. PMID- 10374209 TI - [Sjogren syndrome and hepatitis C virus infection]. PMID- 10374210 TI - [Spinal cord compression by primary vertebral amyloidoma of lumbar region]. PMID- 10374211 TI - An historical perspective of natural and surgical menopause. PMID- 10374212 TI - Tibolone and the serum lipid/lipoprotein profile: does this have a role in cardiovascular protection in postmenopausal women? PMID- 10374213 TI - Symptoms among midlife women: cultural lenses, research, and health care. PMID- 10374214 TI - Two-year prospective and comparative study on the effects of tibolone on lipid pattern, behavior of apolipoproteins AI and B. AB - OBJECTIVE: To investigate long-term lipid and lipoprotein changes in postmenopausal women treated with tibolone in a prospective study using appropriate control groups. DESIGN: Seventy-six of 105 postmenopausal women initially selected for this study completed the 2-year follow-up. Patients were allocated into three groups. The first received 2.5 mg/day tibolone continuously (n = 27; group T), the second received 0.625 mg/day conjugated equine estrogen plus 2.5 mg/day of medroxyprogesterone (group E-P) continuously (n = 25), and a third group contained an additional 24 women who did not receive replacement therapy; these constituted the untreated control group (group C). Plasma lipids and lipoproteins were determined in all patients before joining the study and also at 12 and 24 months after being included. RESULTS: Women treated with tibolone experienced the greatest decreases in cholesterol, both total and high density lipoprotein (HDL), and triglycerides (TG), whereas the highest increase in HDL was observed in the group E-P. A decrease in low density lipoprotein levels was detected in both therapy groups, whereas a significant increase was observed in the control group. TG were increased after E-P therapy. In all the groups, apolipoprotein AI showed parallel trends to HDL and apolipoprotein B to low density lipoprotein. CONCLUSIONS: Both therapy groups, tibolone and E-P, induced changes in levels of plasma lipids, lipoproteins and apolipoproteins. Long-term tibolone treatment is associated with a marked and significant decrease in HDL apolipoprotein AI and TG, an effect that defines the major difference with standard HRT. Clearly, further studies are necessary to establish the definite risk/benefit ratio of tibolone with respect to its overall effect on lipid metabolism. PMID- 10374215 TI - Effects of tibolone on serum lipoprotein and apolipoprotein levels compared with a cyclical estrogen/progestogen regimen. AB - OBJECTIVE: The purpose of the study was to examine the effects of tibolone, a synthetic steroid that alleviates climacteric symptoms and prevents bone loss without inducing monthly bleeds, on lipoprotein cardiovascular risk markers and to compare the effects with those of a standard combined estrogen/progestogen preparation. DESIGN: Ninety-seven postmenopausal women were randomly allocated to receive either tibolone 2.5 mg/day or conjugated equine estrogens 0.625 mg/day with norgestrel 0.15 mg/day for 12 of each 28 days. Fasting serum levels of lipids, lipoproteins, and apolipoproteins (Apo) were monitored during 18 months of treatment. Women on the cyclical preparation had levels determined during both estrogen-only and combined phases. RESULTS: Tibolone caused reductions in triglycerides (33%, p < 0.001), very low density lipoprotein (VLDL) cholesterol (43%, p < 0.001), and high density lipoprotein (HDL) cholesterol (18%, p < 0.001). The HDL2/HDL3 ratio fell by 22% (p < 0.001). Levels of Apo AI and AII were reduced by 18 and 8%, respectively (p < 0.001). The combined preparation caused reductions in VLDL cholesterol (23%, p < 0.001) and low density lipoprotein cholesterol (15%, p < 0.001). There were small reductions in HDL3 cholesterol and in Apo AII and Apo B. All parameters, except for Apo AII and Apo B and lipoprotein (a) [Lp (a)], showed cyclical changes. Lp (a) levels were reduced significantly by both treatments. CONCLUSIONS: The cyclical preparation had potentially beneficial effects on LP risk markers. The reduction in HDL induced by tibolone constitutes a potentially adverse change, which may be offset by the substantial falls in triglycerides, VLDL cholesterol, and Lp (a). PMID- 10374216 TI - Symptom experience of Filipino American midlife women. AB - OBJECTIVE: The purpose of this study was to describe the perimenopausal symptom experience of Filipino American midlife women with particular emphasis upon estrogen-related menopause symptoms (day sweats, hot flashes, night sweats, and vaginal dryness). DESIGN: A cross-sectional, descriptive survey was used to generate symptom experience data for 165 Filipina Americans between the ages of 35 and 56 who self-identified as Filipina American and were English-language proficient. The community-based sample completed questionnaires composed of sample characteristic questions and a 51-item menstrual symptom checklist with menopause-related symptoms embedded in it. RESULTS: Sample characteristics and symptom experience were compared among age groups of 35 to 39 (n = 39), 40 to 44 (n = 40), 45 to 49 (n = 37), and 50 to 56 (n = 49) and by perimenopausal phase, defined as premenopausal (n = 85), transitional (n = 33), and menopausal/postmenopausal (n = 47). The most reported individual symptoms were "felt energetic" (86.1%) and "well-being" (83.6%). Estrogen-related menopause symptoms were reported as "vaginal dryness" (39.4%), "hot flashes" (37.6%), "day sweats" (27.9%), and "night sweats" (24.2%) by the total sample. Distress associated with estrogen-related menopause symptoms was reported by 17% (n = 28) of all women. Subjects' chi 2 tests indicated that 50-to-56-year-old women were more likely to report fatigue/sleep symptoms, physical symptoms, and estrogen related menopause symptoms than all other age groups. When compared by perimenopausal phase, transitional women were more likely to report moderate or extreme severity for day sweats. Premenopausal women were more likely to report minimal or mild severity and women in the perimenopausal transition were more likely to report moderate or extreme severity on estrogen-related menopause symptoms. CONCLUSIONS: Filipino American midlife women appear to consider the perimenopausal transition in a positive light and experience little distress associated with estrogen-related menopause symptoms experienced. Findings from this study suggest that Filipina Americans view perimenopausal symptoms as part of a normal life stage that does not warrant concern. PMID- 10374217 TI - Symptom responses of midlife Filipina Americans. AB - OBJECTIVE: The purpose of this study was to describe the perimenopausal symptom responses of Filipino American midlife women. DESIGN: This cross-sectional, descriptive survey collected symptom response information on Filipino American midlife women aged 35 to 56 years (n = 165) who self-identified as Filipina American and were proficient in the English language. Women were recruited from community churches and social groups. A survey questionnaire comprised of health history questions and a symptom checklist with symptom response questions were completed by the participants. RESULTS: The symptom responses of women were compared by age groups of 35 to 39 (n = 39), 40 to 44 (n = 40), 45 to 49 (n = 37), and 50 to 56 (n = 49) and by perimenopausal phases of premenopausal (n = 85), transitional (n = 33), and menopausal/postmenopausal (n = 47). Indications from chi 2 tests showed that women in the 35-to-39- and 50-to-56-year groups were more likely to take acetaminophen (Tylenol) or aspirin for symptoms, and women in the 45-to-49- and 50-to-56-year groups were more likely to be on hormone replacement therapy. Surprisingly, women in the 50-to-56-year group were less likely to use talking with friends as a symptom management strategy. CONCLUSIONS: Nonpharmacologic symptom management strategies exceeded the use of medications (hormones, over-the-counter) by Filipina Americans. This may be a strong indicator of their positive attitude toward this phase in their life and sends a message to clinicians about the choices that these women make for symptom management. Culturally relevant care would include alternatives to hormone replacement therapy in education materials about the perimenopausal transition for midlife Filipinas. PMID- 10374218 TI - The North American Menopause Society 1998 Menopause Survey. Part I: Postmenopausal women's perceptions about menopause and midlife. AB - OBJECTIVE: To collect information relevant to the mission of The North American Menopause Society (NAMS)--i.e., increasing understanding of menopause--by assessing perceptions held by postmenopausal women in the United States aged 50 to 65 years regarding their menopause transition and early postmenopausal years. DESIGN: During the period from June to July 1998, The Gallup Organization conducted 752 telephone interviews with a randomly selected sample of postmenopausal women aged 50 to 65 years from across the United States, based on questions developed by NAMS. In Part I of this survey, women were asked about their personal experiences with menopause, their health-related lifestyle changes since premenopause, their frequency of discussing menopause, and their rating of preparedness for menopause. Part II of this survey, including use of hormone replacement therapy as well as use of healthcare services, will be reported in a future communication from NAMS. RESULTS: The majority (51%) of the postmenopausal women surveyed reported being happiest and most fulfilled between the ages of 50 to 65 years, compared with when they were in their 20s (10%), 30s (17%), or 40s (16%). Many areas of their lives had improved since menopause, including family/home life, sense of personal fulfillment, ability to focus on hobbies or other interests, relationship with spouse/partner, and friendships. A majority (51%) said their sexual relationships had remained unchanged. Approximately three quarters of women surveyed reported making some type of health-related lifestyle change, such as stopping smoking, at menopause/midlife. Women who had undergone hysterectomy expressed more improvement than women with an intact uterus, especially in the areas of sexual relationships, spouse/partner relationships, personal fulfillment, and physical health; data are not available regarding the health state of these women before surgery or whether they experienced surgical menopause, but this improvement did not appear to be the result of hormone replacement therapy. Women tended to look to women from their own generation for menopause-related information and believed that they have prepared the younger generation for menopause better than they were prepared by their mothers' generation. Those surveyed advised younger women to engage in healthful activities and become knowledgeable so that they could make informed health decisions. CONCLUSIONS: Although the postmenopausal women surveyed had differing views of menopause as well as their perceptions of postmenopause compared with premenopause, the majority viewed menopause and midlife as the beginning of many positive changes in their lives and health. Hysterectomy was a factor associated with improved sexual relationships, spouse/partner relationships, sense of personal fulfillment, and physical health. PMID- 10374219 TI - Menopause-related oral alveolar bone resorption: a review of relatively unexplored consequences of estrogen deficiency. AB - OBJECTIVE: The alveolar processes of the maxilla and mandible provide the bony framework for tooth support. Osteoporotic changes of these bones may directly affect tooth stability and retention. This report reviews studies that have evaluated the relationship between systemic osteoporosis and oral alveolar bone mass as well as the effect of estrogen use on oral alveolar bone and tooth retention. DESIGN: Ten years (1989-1998) literature review. RESULTS: Studies reviewed demonstrate a positive correlation between systemic bone mass and systemic osteoporosis to oral bone resorption. Estrogen replacement therapy affects oral bone in a manner similar to the way it affects other sites. CONCLUSIONS: It is evident that postmenopausal estrogen users may retain more teeth after menopause. Sustained oral health and better tooth retention are potentially additional benefits for hormone replacement therapy users after menopause. PMID- 10374220 TI - Raloxifene lowers serum lipoprotein(A) in healthy postmenopausal women: a randomized, double-blind, placebo-controlled comparison with conjugated equine estrogens. AB - OBJECTIVE: Long-term postmenopausal estrogen replacement therapy lowers the risk of osteoporotic fractures and coronary artery disease but increases the risk of endometrial cancer and probably breast cancer. Raloxifene, a nonsteroidal estrogen receptor ligand, seems to have a tissue-specific antiestrogenic action on endometrium and breast and the desired estrogenic action on bone and lipid metabolism. The purpose of this study was to investigate the effects of 24-month treatment with orally administered raloxifene in two doses (60 mg and 150 mg daily) and conjugated equine estrogens in a standard oral dose (0.625 mg daily) on serum lipoprotein(a) [Lp(a)], an independent risk factor for coronary artery disease, in healthy postmenopausal women who had undergone hysterectomy. DESIGN: A randomized, double-blind, placebo-controlled study was performed with 56 women. RESULTS: In the placebo group serum Lp(a) levels did not change throughout the study. After 6 months, serum Lp(a) was significantly reduced versus baseline in the raloxifene 150 (-17%; p = 0.003) and conjugated equine estrogens (-26%; p = 0.003) groups, but this reduction was significantly different from placebo only in the conjugated equine estrogens group. At 12 and 24 months, serum Lp(a) levels were significantly lowered versus baseline in all active treatment groups. However, these reductions were significantly different from placebo only in the raloxifene 150 and conjugated equine estrogens groups. After 24 months, serum Lp(a) was reduced versus baseline with 30% (p = 0.001) in the raloxifene 150 group and 35% (p = 0.001) in the conjugated equine estrogens group. CONCLUSIONS: Long term raloxifene treatment significantly lowers serum Lp(a) levels in postmenopausal women and thus might reduce the risk of coronary artery disease. PMID- 10374221 TI - Differential effects of estrogen-androgen and estrogen-only therapy on vasomotor symptoms, gonadotropin secretion, and endogenous androgen bioavailability in postmenopausal women. AB - OBJECTIVE: To investigate somatic symptom relief, gonadotropin secretion, and endogenous androgen bioavailability (protein-bound and free) during 3 months of estrogen-androgen therapy or matched estrogen-only replacement therapy. DESIGN: Ninety-three naturally menopausal outpatients with 6 or more months of amenorrhea, who were experiencing mild-to-moderate vasomotor symptoms, were randomized to receive one of five treatments: oral esterified estrogens (0.625 mg or 1.25 mg), oral esterified estrogens combined with methyltestosterone (0.625 mg combined with 1.25 mg methyltestosterone or esterified estrogens 1.25 mg combined with 2.5 mg methyltestosterone), or placebo for 12 weeks. All treatments were preceded by a 4-week placebo lead-in period. RESULTS: Patients receiving the lower dose of estrogen-androgen therapy had fewer somatic menopausal symptoms than patients receiving the lower dose estrogen (0.625 mg), and they experienced somatic symptom relief similar to those patients receiving the higher dose of estrogen (1.25 mg). Significantly greater luteinizing hormone suppression (p < or = 0.03) occurred in estrogen-androgen groups compared to estrogen groups, suggesting that added androgen might mediate a more pronounced negative feedback on the hypothalamic-pituitary axis. Sex hormone-binding globulin increased significantly in both estrogen-treated groups (p < or = 0.01), whereas decreases occurred in both estrogen-androgen groups (p < or = 0.006). The higher dose estrogen-only preparation significantly reduced androstenedione (p < or = 0.01) and dehydroepiandrosterone sulfate (p < or = 0.005). CONCLUSION: The extent of relief with lower dose estrogen-androgen therapy was similar to higher dose estrogen-only treatment. The greater efficacy of combination therapy on somatic symptoms could be mediated by the same mechanism responsible for the suppressive effects of estrogen-androgen therapy on luteinizing hormone secretion. The marked differences in circulating levels of sex hormone building globulin, which were increased by estrogen and decreased by estrogen-androgen, and the resulting impact on bioavailable androgens and estrogens could also explain the differential somatic relief with both treatments. Endogenous adrenal androgens were lower in women treated with esterified estrogens 1.25 mg/day, suggesting that estrogen therapy can produce a significant hypoandrogenic state by inhibiting production or accelerating clearance of adrenal androgens. PMID- 10374223 TI - Effect of initial gestagen treatment on bleeding patterns in postmenopausal women receiving continuous combined hormone replacement therapy. AB - OBJECTIVE: The purpose of this prospective study was to investigate the association of initial gestagen treatment with future breakthrough bleedings during continuous combined hormone replacement therapy. The predictability of progesterone challenge test on bleeding has also been investigated. DESIGN: Eighty-six naturally postmenopausal women, of whom 38 received initial gestagen treatment (5 mg medroxyprogesterone acetate, twice daily, for 10 days) and 48 did not, were included in this prospective study. Patients were followed for 6 months, and any bleeding occurring during therapy was recorded. RESULTS: Of the 48 patients who received continuous combined hormone replacement therapy without initial gestagen treatment, 23 (48%) had a bleeding episode. Of the 38 patients who received the same therapy with initial gestagen treatment, 13 (34.2%) had bleeding. There was a trend toward decreased breakthrough bleeding in the second group, which did not achieve significance. The mean time before bleeding was 6.76 weeks in the first group and 11.75 weeks in the second group; the difference was statistically significant (p < 0.05). Of the 28 patients with a negative progesterone challenge test at the onset of therapy, eight (28.6%) had bleeding during hormone replacement therapy. Of the 10 patients with a positive challenge, five (50%) had bleeding. The difference was not statistically significant. CONCLUSIONS: Patients who receive gestagen treatment before the onset of continuous combined hormone replacement therapy tend to experience breakthrough bleeding less frequently and later during the therapy. The response to progesterone challenge does not predict future bleeding during continuous combined hormone replacement therapy. PMID- 10374222 TI - Hormone replacement therapy for African American women: missed opportunities for effective intervention. AB - OBJECTIVES: Because of the potential benefits and risks of hormone replacement therapy (HRT), information about the efficacy of HRT in different groups of women is important to patients and providers. The objectives of this study were to review the evidence on the benefits and risks of HRT in African American women and to present a quantitative analysis of the potential reduction in mortality from osteoporotic fractures and coronary heart disease and the potential increase in risk of breast and endometrial cancer. METHODS: A MEDLINE search of English language observational studies and clinical trials on the effects of HRT on osteoporotic fractures and coronary heart disease (CHD) was conducted for the time period from 1966 to September 1998. Using available CHD mortality data for African American women and white women, potential reductions in mortality with HRT were explored for African American and white women. RESULTS: In the 30 studies on CHD and HRT, African American women were known to comprise only 173 (0.1%) of 148,437 participants. In 11 studies of HRT and osteoporotic fractures, only 128 (0.4%) of 40,299 participants were known to be African American women. An analysis of CHD mortality by decade intervals indicated that African American women, aged 55 to 64, are more likely to die from CHD each year than white women. Despite a lower incidence of breast and endometrial cancer among African American women, the mortality rates of African American women with these cancers is higher compared with white women. CONCLUSIONS: With the higher underlying CHD mortality rate among African American women, HRT is an important potential preventive therapy. The absence of African American women and other non-white women from clinical studies of HRT makes it difficult to fully assess the risks and benefits of HRT in this group of women. PMID- 10374224 TI - Perceived barriers and recommendations concerning hormone replacement therapy counseling among primary care providers. AB - OBJECTIVE: To increase our understanding of the factors that impede or promote counseling about hormone replacement therapy, we asked clinicians to provide information concerning barriers and strategies to promote counseling. DESIGN: We asked clinicians to consider two different scenarios: (1) what they do in they current practice and (2) what they would do if their health care systems implemented the United States Preventive Services Task Force recommendation regarding hormone replacement therapy counseling. A total of 49 of 50 invited clinicians participated in one of six focus group interviews (three women's groups and three men's groups). Our analysis consisted of four steps: (1) identifying segments and classifying them into themes, (2) categorizing themes into topic areas, (3) establishing a final consensus of themes and topics, and (4) ascertaining similarities and contrasts among groups. Transcripts of sessions were analyzed across groups for themes using a text-based analysis system. Conceptualization of themes was derived using a system model of preventive care. Interrater agreement before consensus was good: Kappa (kappa) ranged from 0.70 to 1.00. RESULTS: For current practice, identified barriers included lack of information about risks and benefits, unique challenges of counseling, and lack of resources to conduct counseling. The major strategies suggested were to develop and distribute patient education materials. Discussions about barriers to implementing the United States Task Force recommendation focused on lack of information and resources. CONCLUSIONS: Suggested strategies were multiple, involving individual-, relationship-, and system-level interventions. We expect the strategies identified to be supportive of future efforts to promote counseling for hormone replacement therapy. PMID- 10374225 TI - Anticipating menopause: observations from the Seattle Midlife Women's Health Study. AB - OBJECTIVE: The purpose of this study was to determine midlife women's images of menopause and their expectations of their own menopausal experiences. DESIGN: Participants in the Seattle Midlife Women's Health Study (n = 508) responded to a question about their definitions of menopause, and their expectations and concerns about their own menopausal experiences during an in-person interview conducted at entrance to the study between late 1990 and early 1993. At that time, women ranged in age from 35 to 55 years (median, 41 years); 80% were European American and were well educated (median, 15 years). RESULTS: Women defined menopause in the following ways: (1) cessation of their periods, (2) end of their reproductive ability, (3) a time of hormonal changes, (4) a change of life, (5) a changing body, (6) changing emotions, and (7) an aging process. Few women defined menopause as a time of symptoms or disease risk or a time for medical care. Women were most likely to be uncertain of their expectations of their own menopause, and many had no expectations. CONCLUSIONS: This cohort of midlife women did not seem to have adopted a medical model of menopause and were most likely to view menopause as a normal developmental process. Their uncertainty about what to expect provides an opportunity for health teaching and anticipatory guidance. PMID- 10374226 TI - Age at menopause in women with type 2 diabetes mellitus. AB - OBJECTIVE: Considering that chronic diseases such as diabetes mellitus (DM) may determine premature ovarian failure by various mechanisms, we studied the age at menopause in women without diabetes and in women with type 2 DM. DESIGN: We studied 409 women without diabetes and 404 patients with type 2 DM, selected from 45 to 55 years of age, for analysis with the status quo method. The age at menopause was calculated with a logistic regression on the proportions of menopausal women for each age group. RESULTS: In the groups, 172 women without diabetes and 207 women with diabetes had menopause. The regression procedure gave a median age of 49.7 +/- SD 1.7 years for the whole group, 49.6 +/- 1.6 years for the nondiabetic group, and 49.8 +/- 1.7 years for women with diabetes. Women without diabetes were 1.4 years younger, but this factor did not have an influence on the results. Smoking habits, vegetarianism, and somatometric variables were similar in both groups, except for waist/hip and abdomen/hip ratios, larger in the group of women with diabetes. The mean for years since diagnosis in patients < 45 years of age was 4.9 years. For older patients, the figure increased to 8.9 years. CONCLUSIONS: No difference for age at menopause was found between women without diabetes and women with type 2 diabetes who were 5 to 8 years since the diagnosis was made. PMID- 10374227 TI - Helping women achieve long-term continuance of estrogen replacement therapy (ERT) and hormone replacement therapy (HRT) PMID- 10374228 TI - Hysterectomy, ovarian failure and depression. PMID- 10374229 TI - [Results of cervical interbody fusion with coral grafts]. AB - OBJECTIVES: We present the long term clinical and radiological results of a retrospective series of 46 cervical interbody fusions using coral grafts performed in 38 patients. MATERIAL AND METHODS: The patients were treated for prolapsed discs (19 cases) or cervical spondylosis (19 cases) with a clinical presentation of either radiculopathy (31 cases) or myelopathy (7 cases). We have done a post-operative clinical analysis of cervicoscapulalgia and radiculo medullary symptoms and a radiological comparison of the change of the cervical spine angulation, the loss of height and the fusion rate at the graft site. RESULTS: The early clinical postoperative outcome showed that 10 out of 20 patients with excellent radiculo-medullary results had cervicoscapulalgia and 13 out of 18 patients with partial improvement had cervicoscapulalgia. No poor results according to our classification were noted. The late clinical outcome (average follow-up of 16 months) showed that 15 patients out of 31 had radiculo medullary degradation and 24 presented with cervicoscapulalgia. Sixteen out of 20 patients had a loss of lordosis (range 6.2 degrees; SD 1.2) and 17 a loss of height (range 11.3%; SD 1.5). After 2 years, 13 out of 20 grafts were still hyperdense compared to the adjacent bone, and 8 had a hypodense peripheral edge. CONCLUSIONS: Coral grafts of this series have not been able to keep a physiological sagittal balance of the cervical spine, which is probably one of the essential factors to prevent postoperative cervicoscapulalgia. In the same way, the loss of height of the fused segments, by narrowing of the intervertabral foramen, may explain some further radiculo-medullary deterioration. PMID- 10374230 TI - [Tentorial meningioma. Surgical experience with 20 cases]. AB - We report our experience and long-term results of twenty patients with tentorial meningiomas who underwent surgical removal between 1987 and 1996. Computed tomography, angiography and magnetic resonance imaging were used as diagnostic tools for planning the surgical procedure. The tumor site was posterolateral in 6 cases (30%), posteromedial in 4 cases (20%), in the tentorium itself in 4 cases (20%), anterolateral in 3 cases (15%), at the apex of tentorial incisura in 2 cases (10%) and at the free border of the tentorial notch in 1 case (5%). Neuroradiologically, 70% of the meningiomas ranged from 1 to 3 cm. Lateral and medial tumors with solely or mainly supratentorial development were approached from above. The approach from below was selected for meningiomas with subtentorial involvement only. In meningiomas with both supra and subtentorial growth, a supratentorial bone flap was combined with a suboccipital craniectomy using a retromastoid incision. Radical surgical removal (Simpson's grade I and II) was achieved in 80% of the cases. There was no mortality. The follow-up averaged 4 years and revealed that 65% of patients were able to return to their premorbid activity. Complications were mainly postoperative brain oedema, functional deficits, seizures and psychological disorders. Recurrence rate amounted at 6.25% in the group where the tumors were totally removed (16 cases). From this retrospective study, the statistically significant prediction of a good outcome was: duration of symptoms from onset to the operation inferior or equal to 1 year (p < 0.01), good preoperative neurological conditions (Karnofsky scale from 80 to 100) (p < 0.05) and tumor size inferior or equal to 3 cm (p < 0.002). PMID- 10374231 TI - [Temporary occlusion in surgical management of intracranial aneurysm. Report of 54 cases]. AB - Temporary arterial occlusion (TAO) is commonly used in the surgery of intracranial giant aneurysms. Its usefulness and safety in the surgical management of all cases of aneurysms remains to be proved. We report a series of 54 patients operated on for an intracranial aneurysm with the use of TAO. Among the 27 patients, admitted before the 4th day following post subarachnoid hemorrhage with I or II on WFNS score clinically, 24 had early aneurysm surgery. The size of the aneurysm was small in 16 cases, medium in 22, large in 13 and giant in 3 cases. The protocol proposed by Batjer in 1988 for large and giant aneurysms (etomidate, normotention and hypervolemia) was used without any electrophysiological monitoring. All patients underwent a post-operative cerebral CT scan to evaluate the incidence of a cerebral ischemia. Serial transcranial doppler was used to evaluate the severity of vasospasm. Clinical results were assessed using the GOS. TAO was elective in 51 patients and done after peroperative aneurysm rupture in 3 patients. The duration of TAO was less than 5 mn in 25 patients, between 5 and 10 min in 12, between 10 and 15 in 11, between 15 and 20 in 5 and more than 20 min in one patient. The last one developed a reversible neurological deficit secondary to ischemia attribuated to TAO. Intracranial aneurysm peroperative rupture was noted in 3 patients, clinical vasospam in 13 patients. These results allow us to recommend the routine use of TAO in the surgery of intracranial aneurysm. When application time is limited and cerebral protection used, TAO is safe. It decreases the risk of intraoperative rupture from a 18% rate in literature to 4.2% in our present experience and the risk of symptomatic vasospasm is not increased. PMID- 10374232 TI - [Cauda equina tumors in adults]. AB - Large series of cauda equina tumors in adults are seldom reported. This study was based on the 231 cases collected in the French neurosurgery units for the congress of the Societe Francaise de Neurochirurgie in October 1996. Schwannomas were the most frequent benign tumor in this series, followed by ependymomas. Very few malignant tumors were recorded, usually malignant neurinomas nearly always arising in patients with neurofibromatosis. Some other rare tumors were also observed including paragangliomas. This series confirms the contribution of pre therapeutic neurological status to functional prognosis. All schwannomas can be cured while ependymomas and paragangliomas may recur after a very long delay. Surgery must be as complete as possible since adjuvant therapies have proven to have little efficacy. This type of tumor requires a very long follow-up. Prognosis is good for hemangioblastoma. When present, abnormal sphincter function is an argument for poor prognosis. It may appear after primary surgery or more often after treatment of recurrence. PMID- 10374233 TI - [Central neurocytoma. A report of 4 cases]. AB - We report 4 cases of central neurocytoma removed by a transfrontal approach with no recurrence after a mean follow up of 3 years. This uncommon lesion of the supratentorial ventricles (150 cases reported) occurs in young adults, and often induces intracranial hypertension secondary to an obstructive hydrocephalus. The CT scan, MRI and histopathological features are related. This neuronal origin tumor is difficult to distinguish from other intraventricular processes as oligodendroglioma or ependymoma. However, the immunopositivity for the neuronal markers as synaptophysin, calcineurin and microtubul associated protein 2, and the negativity for the glial fibrillary acidic protein, allow the diagnosis of this neuropathological entity. The prognosis is favorable though some cases of recurrence (14 cases) and cerebrospinal dissemination (2 cases) has been reported in the literature. PMID- 10374234 TI - [A symptomatic choroid plexus cyst in the lateral ventricle]. AB - Choroid plexus cyst is generally small and a relatively common finding at autopsy. Huge and symptomatic cysts are rare. Few cases are reported in the literature. We report one case of symptomatic choroid plexus cyst of the right lateral ventricle in a six month baby who presented with epilepsy. Cerebral CT scan and MRI showed a large cyst in the right lateral ventricle compressing the adjacent structures. Total removal of the cyst has been performed by a parieto temporal approach. The course was uneventful. PMID- 10374235 TI - [Cervical myelopathy from ossification of the posterior longitudinal vertebral ligament. Report of 2 cases]. AB - BACKGROUND AND PURPOSE: Cervical myelopathy due to an ossification of the posterior longitudinal ligament (OPLL) is a rare entity in western countries but frequent in Japan. We report on two Lebaneese patients aged 67 and 72 years respectively, who were twins and presented with OPLL. METHODS: Diagnosis was made on myelography in the first case (1989) and on MRI of the cervical spine in the second case (1994). RESULTS: A wide laminectomy was performed in the first case followed by a marked improvement. In the second case, corporectomy of the third, fourth and fifth vertebra with removal of the ligament followed by bone graft didn't improve the clinical symptoms. CONCLUSION: The cause of OPLL remains unknown: genetic factors and metabolic abnormalities are outlined. Treatment options are discussed. PMID- 10374237 TI - [Epidermoid cyst in the occipital skull vault. A case report]. AB - We report a case of an epidermoid cyst of the occipital skull vault in a 28 year old woman. The symptomatology was an occipital swelling and pain. The diagnosis was evoked on the skull X-ray and computed tomography scan. Total removal of the tumor provided complete recovery. PMID- 10374236 TI - [Intramuscular myxoma. A case of myxoma of the spinal erector muscle]. AB - A case of intramuscular myxoma, extended in the lumbar erector spinae muscles, is reported. This soft tissue tumor is rare, about one hundred observations are indexed in the literature. The clinical findings are nonspecific, magnetic resonance imaging reveals few characteristics, and finally a histological examination is required, after surgical removal, to pinpoint the diagnosis. Recurrence is uncommon, but a systematic follow-up is necessary. PMID- 10374238 TI - [Recent progress in akathisia]. AB - Neuroleptic-induced akathisia should be definitely diagnosed as acute, tardive, withdrawal, and chronic. The diagnostic assessment must be identified from the subjective report and objective features. Various assessments of measuring akathisia can be clinically used by instrumental methods and rating scales. The pharmacological basis of neuroleptic-induced akathisia is the inhibition of the dopamine receptors in the brain. The pathogenesis of neuroleptic-induced akathisia may involve GABAergic hypoactivity, noradrenergic hyperactivity, and serotonergic dysfunction in CNS. Iron deficiency and hyperglycemia may be risk factors of neuroleptic induced akathisia in relation to the dopamine function in the brain. Neurological disorders may be associated with the development of a syndrome resembling drug-induced akathisia. The lesion of the thalamic nuclei would originally produce the syndrome. The difference between acute and tardive akathisia on the strategy of the drug treatment should be sufficiently comprehended. In particular, the long-term use of anticholinergic drugs and benzodiazepines should not be prevailed. PMID- 10374239 TI - [Biological rhythms and drugs]. AB - Precise, rhythmic, daily change of the internal milieu is a conspicuous feature of all living organisms. It affects the temporal patterns of all kinds of behaviors during a day and deeply influences both the social structure and daily life of individual human beings. These daily variations arise from the internal circadian mechanisms. Three functions of the endogenous clock are discriminated; rhythm generation, entrainment to light-dark cycle and output from the clock. The endogenous clock is localized in the suprachiasmatic nucleus in mammals. Interestingly, recent papers demonstrated the widespread expression (hippocampus, cerebellum and also peripheral body) of clock genes such as mPer1 and mPer2 (homologs of the Dorsophila period gene). In this review, I describe the cellular mechanisms of circadian oscillation, entrainment and output. Serotonergic drugs and melatonin affect the light and non light-induced entrainment, and lithium and estrogen change the oscillation. Thus drugs can regulate the circadian oscillation, entrainment and outputs. Therefore, I mention the possibility that CNS drugs are useful to keep physiological homeostasis through their action on biological rhythms. PMID- 10374240 TI - [Effects of demeclocycline on psychiatric polydipsia in schizophrenic patients]. AB - Excess intake of water by schizophrenic patients is referred to as psychiatric polydipsia. This symptom causes incontinence, vomiting and hyponatremia, and may sometimes lead to death. We have no effective therapeutic methods other than administrating sodium chloride solution and diuretics, or restricting the intake itself. A case was reported stating that demeclocycline, used in case where there is the syndrome of inappropriate secretion of antidiuretic hormone (SIADH), was effective for the treatment of psychiatric polydipsia. We administered demeclocycline to 8 schizophrenic patients with psychiatric polydipsia, and noticed improvement in incontinence, vomiting and hyponatremia. There was also a decrease of polydipsic behavior. Demeclocycline inhibits the antidiuretic effect of vasopressin on the distal renal tubule. Considering the function of demeclocycline and the relevance of vasopressin to the central nervous system, it has been suggested that demeclocycline has effects on the central nerve through vasopressin or cyclic AMP. PMID- 10374241 TI - [Effect of low-dose sulpiride on dopamine metabolism in rat nucleus accumbens and locomotor activity: studies with intracerebral dialysis]. AB - Changes in the concentration of dopamine (DA) and its metabolites, 3, 4 dihydroxyphenyl-acetic acid (DOPAC) and homovanillic acid (HVA), were determined sequentially in freely moving rats by using a brain dialysis method. The purpose of this study was to investigate the relationship between changes in the dopamine metabolism in the nucleus accumbens and locomotor activities following low-dose (5 mg/kg) sulpiride administration. The DA content in the dialysis fluid rose significantly 4 to 6 h after sulpiride administration. The DOPAC content rose significantly 6 h after sulpiride administration. The HVA content did not show a significant change. A slight increase of locomotor activity was found, but it was not statistically significant. The elevation of DA release in nucleus accumbens by sulpiride might relate to the clinical efficacy of low-dose administration of this drug for mood disorders. PMID- 10374243 TI - Inhibitory effect of some triterpenes from cacti on 32Pi-incorporation into phospholipids of HeLa cells promoted by 12-O-tetradecanoylphorbol-13-acetate. AB - Seventeen triterpenes isolated from cacti and the 10 derivatives were examined for the inhibition of tumor promoter-induced effects in vitro, such as stimulation of 32Pi-incorporation into phospholipids of cultured cells. Betulinic acid (1), cochalic acid (15), erythrodiol (16), oleanolic acid (21) and queretaroic acid (24) inhibited 12-O-tetradecanoylphorbol-13-acetate (TPA) stimulated 32Pi-incorporation into phospholipids of the cultured cells. PMID- 10374242 TI - Possible involvement of membrane phospholipids in inhibition by ferrous ions of [3H]MK-801 binding to the native N-methyl-D-aspartate channel in rat brain. AB - Prior treatment with ferrous chloride led to the marked inhibition of [3H](+)-5 methyl-10, 11-dihydro-5H-dibenzo[a, d]cyclohepten-5, 10-imine (MK-801) binding to an open ion channel associated with the N-methyl-D-aspartate (NMDA) receptor in a concentration-dependent manner at concentrations of higher than 1 microM in rat brain synaptic membranes. Both phospholipases A2 and C significantly prevented the inhibition when treated before the treatment with ferrous chloride, while neither superoxide dismutase nor alpha-tocopherol affected the inhibition even when treated simultaneously with ferrous chloride. Of various saturated and unsaturated fatty acids, moreover, both oleic and arachidonic acids exclusively decreased the potency of ferrous chloride to inhibit binding when membranes were treated with fatty acids, followed by a second treatment with ferrous chloride. These results suggest that membrane phospholipids may be, at least in part, responsible for the interference by ferrous ions in the opening processes of the native NMDA channel through molecular mechanisms associated with the release of unsaturated fatty acids in rat brain. PMID- 10374244 TI - Inhibitory effect of resveratrol on interleukin 6 release by stimulated peritoneal macrophages of mice. AB - In the present investigation, interleukin 6 (IL-6) activity in the supernatant of cultured mouse peritoneal macrophages was monitored using a sensitive bioassay involving the IL-6-dependent murine hybridoma B9 cell line. The effects of resveratrol on Il-6 release by mouse peritoneal macrophages stimulated with calcium ionophore A23187 and fMLP were explored. Resveratrol, at a concentration range from 5 x 10(-6) to 4 x 10(-5) mol.l-1, was found to dose-dependently inhibit IL-6 release by cultured macrophages induced by A23187 and fMLP, and showed no direct cytotoxic effect, but induced proliferation of cultured mouse thymus cells. Resveratrol, at a concentration range from 10(-8) to 10(-5) mol.l 1, was shown to dose-dependently inhibit calcium ion influx into the cells with the stimulation of fMLP (10(-6) mol.l-1). These results suggest that the blocking of calcium ion influx into cells by reveratrol is one of the possible mechanisms of the IL-6 biosynthesis inhibitory action of resveratrol. PMID- 10374245 TI - Protective effect of swerchirin on hematopoiesis in 60Co-irradiated mice. AB - The protective effect of swerchirin, a purified xanthone isolated from whole herb of Swertia calycina Franch. on hematopoiesis was investigated. A significant increase of colony formation in the spleen (colony forming unit in spleen = CFU S) of mice irradiated with 550 rad 60Co gamma-rays and an enhancement of proliferative response of BMC to rmGM-CSF treated with swerchirin [10 mg/kg, 3 time/wk, i.p.] was observed. After introduction of swerchirin [10 mg/kg, i.p. once] a significant increase in the number of peripheral blood leukocytes and a rise in the serum of colony stimulating factor (CSF) were also confirmed. The types of CSF in serum were M-CSF and other hematopoietic growth factors, which were confirmed using McAb of IL-3, GM-CSF and PcAb of M-CSF. These beneficial effects of swerchirin on hematopoiesis may be related to its activity inducing CSFs and other hematopoietic growth factors, and warrant further evaluation of its usefulness. PMID- 10374246 TI - Anti-emetic principles of Pogostemon cablin (Blanco) Benth. AB - Bioassay-guided fractionation of anti-emetic extracts and constituents of 8 traditional Chinese herbal drugs was performed. Twenty extracts described in Table 1 showed anti-emetic activity on copper sulfate induced-emesis in young chicks. From the n-hexane extract of Pogostemon cablin, patchouli alcohol (1), pogostol (2), stigmast-4-en-3-one (3), retusin (4), and pachypodol (5) were tested and exhibited anti-emetic effects. PMID- 10374247 TI - Effects of qing fei tang (TJ-90) on aspiration pneumonia in mice. AB - The effects of Qing Fei Tang (Sei-hai To in Japanese), a Chinese traditional medical mixture, on aspiration pneumonia were studied using mice inoculated with both Streptococcus pneumoniae and gastric juice as aspiration pneumoniae models. Daily (4 weeks) oral usage of Qing Fei Tang before inoculation reduced remarkably the mortality rate of mice. In this aspiration pneumonia model, xanthine oxidase (XO) activity in the lung tissues was elevated, but this elevation was remarkably decreased by use of Qing Fei Tang. These results suggest that Qing Fei Tang pretreatment can reduce oxygen radical production in inflammed lungs and may reduce the mortality for aspiration pneumonia. PMID- 10374249 TI - Anticonvulsant properties of linalool in glutamate-related seizure models. AB - In order to investigate the pharmacodynamic basis of the previously-established anticonvulsant properties of linalool, we examined the effects of this compound on behavioral and neurochemical aspects of glutamate expression in experimental seizure models. Specifically, linalool effects were investigated to determine its inhibition of (i) L-[3H]glutamate binding at CNS (central nervous system membranes), (ii) N-methyl-D-aspartate (NMDA)-induced convulsions, (iii) quinolinic acid (QUIN)-induced convulsions, and the behavioral and neurochemical correlates of PTZ-kindling. The data indicate that linalool modulates glutamate activation expression in vitro (competitive antagonism of L-[3H]glutamate binding) and in vivo (delayed NMDA convulsions and blockage of QUIN convulsions). Linalool partially inhibited and significantly delayed the behavioral expression of PTZ-kindling, but did not modify the PTZ-kindling-induced increase in L [3H]glutamate binding. PMID- 10374250 TI - Sedative actions of Ternstroemia sylvatica in the male rat. AB - Ternstroemia sylvatica is a plant reputed popularly to possess a anxiolytic properties but has not yet been systematically tested for such activity. The behavioral actions of T. sylvatica were examined using the open field test, the elevated plus-maze test, and the forced swim test in male rats. T. sylvatica (7.1 mg/kg and 14.2 mg/kg, i.p.) reduced ambulatory behavior in the open field test and cancelled the anti-immobility actions produced by desipramine (32 mg/kg, i.p.) in the forced swim test, as did diazepam. In the elevated plus-maze test, T. sylvatica (7.1 mg/kg, i.p.) failed to show anxiolytic actions. It is concluded that Ternstroemia sylvatica produces sedative effects rather than the attributed anxiolytic actions. PMID- 10374248 TI - The traditional Chinese medicine banxia houpo tang improves swallowing reflex. AB - A marked depression of swallowing reflex has been found in patients with aspiration pneumonia. We have examined the effects of Banxia Houpo Tang (BHT, Hange Koboku-To in Japanese), on swallowing reflex among the elderly. Thirty-two patients, mean age 74.2 +/- 1.7 years who had at least one episode of aspiration pneumonia, were divided into two groups. Twenty patients took BHT extracts of 7.5 g per day for four weeks, and the other 12 patients took a placebo. The swallowing reflex was measured by a bolus injection of 1 ml of distilled water into the pharynx through a nasal catheter. The reflex was evaluated by the latency time of response, which was the time from the injection to the onset of swallowing. The latency of response decreased significantly from 11.6 +/- 3.0 sec to 2.6 +/- 0.4 sec in the group treated with BHT (p < 0.01), while in the other group with placebo it was from 11.0 +/- 4.0 to 10.8 +/- 3.6 (p > 0.5). Depletion of substance P in the pharynx causes impairments of the swallowing reflex. Substance P in the saliva of treated patients increased from 9.2 +/- 2.5 fmol/ml to 15.0 +/- 2.2 fmol/ml after BHT treatment (p < 0.01), while levels were 8.0 +/- 4.0 fmol/ml before and 7.1 +/- 3.1 fmol/ml after among the placebo group (no significant difference). We suggest that BHT improves the impaired swallowing reflex and may help to prevent aspiration pneumonia in the elderly. PMID- 10374251 TI - Antiviral activity of sandalwood oil against herpes simplex viruses-1 and -2. AB - Sandalwood oil, the essential oil of Santalum album L., was tested for in vitro antiviral activity against Herpes simplex viruses-1 and -2. It was found that the replication of these viruses was inhibited in the presence of the oil. This effect was dose-dependent and more pronounced against HSV-1. A slight diminution of the effect was observed at higher multiplicity of infections. The oil was not virucidal and showed no cytotoxicity at the concentrations tested. PMID- 10374252 TI - Aged garlic extract attenuates intracellular oxidative stress. AB - Oxidation of low density lipoprotein (LDL) has been recognized as playing an important role in the initiation and progression of atherosclerosis. We recently reported that aged garlic extract (AGE) inhibited LDL oxidation and minimized oxidized LDL-induced cell injury. In this study, the antioxidant effects of AGE were further examined using bovine pulmonary artery endothelial cells (PAEC) and murine macrophages. Lactate dehydrogenase (LDH) release, as an index of membrane injury, and intracellular glutathione (GSH) levels were determined. Oxidized LDL (Ox-LDL) caused an increase of LDH release and depletion of GSH. Pretreatment with AGE prevented these changes. AGE exhibited an inhibition of Ox-LDL-induced peroxides in PAEC. AGE suppressed peroxides in murine Macrophage (J774 cells) dose-dependently. The J774 cells were also incubated with AGE, interferon-gamma (IFN-gamma) and lipopolysaccharide (LPS) and nitric oxide (NO) production was measured. AGE inhibited NO production in J774 cells. In a cell free system, AGE was shown to scavenge H2O2 dose-dependently. Our data demonstrate that AGE can protect the endothelial cells from oxidized LDL-induced injury by preventing depletion of intracellular GSH and by removing peroxides. AGE also reduces levels of NO and peroxides in macrophages. These data suggest that AGE is a useful protective agent against cytotoxicity associated with Ox-LDL and NO, and it may thus be useful for the prevention of atherosclerosis and cardiovascular diseases. PMID- 10374253 TI - Antimicrobial activity of Wedelia trilobata crude extracts. AB - A biological screening of activity against Gram-positive and Gram-negative bacteria, yeasts, and fungi of crude extracts from Wedelia trilobata is reported. The n-hexane extract showed antibacterial activity against Bacillus subtilis, Mycobacterium smegmatis, Staphylococcus aureus, and Staphylococcus epidermidis (Gram-positive bacteria); along with Proteus vulgaris, Pseudomonas aeruginosa, Salmonella group C, Salmonella paratyphi, and Shigella sonnei (Gram-negative bacteria). The ethyl acetate extract was active only against Salmonella group C; and the aqueous extract was inactive against the tested bacteria. None of the tested extracts showed biological activity against the yeasts (Candida albicans, Candida tropicalis, Rhodotorula rubra) or the fungi (Aspergillus flavus, Aspergillus niger, Mucor sp., Trichophyton rubrum). PMID- 10374255 TI - [The 72nd annual meeting of Japan Society for Occupational Health. Tokyo, Japan. May 6-8, 1999. Abstracts]. PMID- 10374254 TI - Seasonal changes in the concentrations of four taxoids in Taxus baccata L. during the autumn-spring period. AB - The concentrations of four common taxoids: baccatin III, paclitaxel, cephalomannine and 10-deacetylbaccatin III (10-DAB III) were measured in fresh needles and stems of Taxus baccata L. during the late autumn-spring period (November'96-April'97) which has not been investigated to date in this species. Baccatin III, paclitaxel and 10-DAB III were present on the surface of the twigs in concentrations of 8-26 micrograms/1000 g (fresh weight). Changes in the levels of baccatin III and paclitaxel inside the needles and stems showed similar trends over the investigated period. From November to March the total level of taxoids differed between the needles and stems, and were the same only in April. Total levels in fresh needles were stable from December to March. The highest concentrations of 10-DAB III in the whole analysed period in fresh stems were measured, as well as in the fresh needles except for samples collected in November and December when the levels of cephalomannine were higher. The concentrations of paclitaxel were usually the lowest. These results confirm that epigenetic factors--date of collection (and thus phyllogenesis) and kind of plant tissue--determine taxoid levels during the late autumn-spring period in T. baccata. The opposite patterns of changes for 10-DAB III and cephalomannine, especially in the fresh needles, suggest a possible role for 10-DAB III in the biosynthetic pathway to cephalomannine, a less polar taxoid with a side-chain at position C-13. As well, owing to the thermolability of taxoids, the influence of low temperatures in December and January could explain the highest observed concentrations of 10-DAB III in the fresh stems and needles, respectively. PMID- 10374256 TI - Ca(2+)-ATPase from chicken (Gallus domesticus) erythrocyte plasma membrane: effects of calmodulin and taurine on the Ca(2+)-dependent ATPase activity and Ca2+ uptake. AB - In the present report we describe a method for purifying plasma membranes from chicken erythrocytes using sonication under conditions that facilitate preferential lysis of plasma membrane, followed by centrifugation through a sucrose gradient. The Ca(2+)-dependent ATP hydrolysis by plasma membranes is activated by nanomolar levels of calmodulin, similarly to that from anucleated erythrocytes. Inside-out vesicles display a calmodulin-activated Ca2+ uptake. Purified Ca(2+)-ATPase is obtained from the plasma membrane by Sepharose calmodulin affinity chromatography, and exhibits an apparent molecular mass of 150 kDa on SDS-polyacrylamide gel electrophoresis, clearly showing that the enzyme is distinct from that described in anucleated erythrocytes (140 kDa). The enzyme is insensitive to physiological concentrations of taurine, a beta-amino acid that has been proposed to be involved in Ca2+ homeostasis of nucleated erythrocytes, suggesting that the effect of taurine is not mediated by the Ca(2+) ATPase. Taken together, these data suggest that the enzyme may be an isoform that resembles the previously described plasma membrane Ca(2+)-ATPase from anucleated erythrocytes in its regulation by calmodulin, but differs in its apparent molecular weight. PMID- 10374257 TI - Effects of UV-C irradiation on phosphoinositide turnover in plant cells: similarities with those occurring via the formation of reactive oxygen intermediates in animal cells. AB - With the aim of examining the response of plant cells to UV-C irradiation, we investigated the behaviour of the phosphatidylinositol 4,5 bisphosphate (PtdIns 4,5-P2) molecule (the precursor of the phosphoinositide signal transduction cascade) by exposing callus cells from Peucedanum verticillare to UV-C (130 J m 2) and by examining the level and the fatty acid composition of PtdIns 4,5-P2 at different times after irradiation. We show that a pathway for the UV-C response includes transient PtdIns 4,5-P2 breakdown. The effect of ultraviolet rays is mimicked by H2O2 suggesting that in this plant it may be brought about by reactive oxygen intermediates (ROI), as already underlined in experimental animal models. PMID- 10374259 TI - Primary structure and function of superoxide dismutase from the ascidian Halocynthia roretzi. AB - A protein with a molecular weight of 17K, immunoreactive with the S-1B2 antibody, has been isolated from hemocytes of Halocynthia roretzi. Its amino acid sequence has been determined by sequential Edman degradation analysis of peptide fragments derived from proteolytic fragmentation. The 17K protein is a single chain protein consisting of 151 amino acids with an acylated N-terminal serine. A comparison of the amino acid sequence of H. roretzi 17K protein with those of other proteins reveals that the 17K protein is Cu,Zn-SOD. The protein was found to have a KCN inhibited SOD activity. Cu,Zn-SOD has been purified from H. roretzi plasma. The molecular weight is 17K and the activity is inhibited with KCN and diethyldithiocarbamate. It has been demonstrated that it can enhance phagocytosis by H. roretzi hemocytes. Thus, plasma Cu,Zn-SOD plays a role in H. roretzi as a defense molecule. PMID- 10374258 TI - Identification, purification and properties of a beta-1,3-glucan-specific lectin from the serum of the cockroach, Blaberus discoidalis which is implicated in immune defence reactions. AB - A lectin specific for laminarin, a beta-1,3-glucan, agglutinating baker's yeast and enhancing prophenoloxidase activation by laminarin, has been purified from the cockroach, Blaberus discoidalis, serum. Purification involved gel filtration with Bio-gel P300 and affinity chromatography on blue Sepharose CL-6B and laminarin-Sepharose 4B. The purified lectin has a molecular mass estimate of 520 kDa determined by gel filtration, and approximately 80 and 82 kDa by SDS-PAGE, under non-reducing and reducing conditions, respectively. After isoelectric focusing the lectin focused as a single band at pH 4.9. The purified lectin was stained by the periodic acid/Schiff's reagent showing that it is a glycoprotein, and was deglycosylated by endo-beta-N-acetylglu-cosaminidase F. Amino acid composition analysis showed the protein is similar to previously purified beta 1,3-glucan binding proteins from other invertebrates. In electron micrographs by negative staining, the protein formed large aggregates with 'Y'-shaped 'structural units' ca. 79 x 65 nm. Immunological tests confirmed that this lectin is not related to any other lectins previously purified from the same insect. This protein appears to be part of the hexamerin family of proteins. This is one of the first reports of a hexamerin-like molecule with lectin activity. PMID- 10374260 TI - Contents of myosin heavy chains in denervated slow and fast rat leg muscles. AB - The total content of myosin heavy chain (MHC) and individual MHC isoforms were studied in 14-day denervated rat leg muscles: the slow-twitch (soleus) and fast twitch (extensor digitorum longus and gastrocnemius) by biochemical methods. The weight of the denervated muscles decreased by about 50%, as compared to the control muscles. In all denervated muscles the total content of MHCs decreased, more so in the slow than in the fast muscles. We have observed that the proportion among the MHC isoforms changed: while MHC-1 and MHC-2B decreased, MHC 2A and MHC-2X increased. Taking into account muscle atrophy, the loss of MHC total content and the shift in pattern of MHC isoforms, the total net changes of the particular MHC isoforms were evaluated. It was found that the muscle content of each of the MHCs decreased after denervation, but their tissue concentration changed variously. The concentration of the MHC-1 and MHC-2B decreased in all denervated muscles, but that of the MHC-2A and MHC-2X changed variously, depending on the muscle. The concentration of MHC-2A decreased in the soleus and increased in the fast muscles, whereas the concentration of the MHC-2X changed inversely. In the denervated soleus a considerable amount of MHC-2X was expressed, while in the contralateral muscles this isoform was undetectable or appeared at trace levels. PMID- 10374261 TI - Dictyostelium ribosomal protein genes and the elongation factor 1B gene show coordinate developmental regulation which is under post-transcriptional control. AB - Starvation for amino acids initiates the developmental program in the cellular slime mold, Dictyostelium discoideum [19, 20]. One of the earliest developmental events is the decline in ribosomal protein synthesis [2, 17, 29, 30]. The ribosomal protein mRNAs are excluded from polysomes with 20 min to 1 h following the removal of nutrients, and their mRNA levels decline sharply at about 9 h into the 24-h developmental cycle [28, 31, 35, 36]. It has been generally assumed that the decline in r-protein mRNA levels during late development reflected a decline in the transcription rate [12, 32, 43]. Here we demonstrate that this is not the case. The transcription rates of three ribosomal protein genes, rpL11, rpL23 and rpS9 as well as an elongation factor 1B gene have been determined during growth and development in Dictyostelium. Throughout growth and development the transcription rate of the ribosomal protein genes remains relatively constant at 0.2%-0.5% of the rate of rRNA transcription while the elongation factor 1B gene is transcribed at 0.4%-0.6% of the rRNA rate. This low but constant transcription rate is in contrast to a spore coat protein gene Psp D, which is transcribed at 6% of the rRNA rate in late developing cells. The elongation factor 1B gene appears to be co-regulated with the ribosomal protein genes both in terms of its transcription rate and mRNA accumulation. Dictyostelium has been a popular model for understanding signal transduction and the growth to differentiation transition, thus it is of significance that the regulation of ribosome biosynthesis in Dictyostelium resembles that of higher eukaryotes in being regulated largely at the post-transcriptional level in response to starvation as opposed to yeasts where the regulation is largely transcriptional [27]. PMID- 10374262 TI - Various stimulators of the cyclic AMP pathway fail to promote adipose conversion of porcine preadipocytes in primary culture. AB - The cyclic adenosine-monophosphate (cAMP) pathway is generally recognized as one of the essential pathways for the adipose conversion of rodent preadipocytes in vitro. However, divergent effects of cAMP on adipocyte differentiation have also been reported. Since there is very little data on non-rodent preadipose cells, the aim of the present work was to analyze the effects of classic activators of the cAMP pathway on the proliferation and differentiation of porcine preadipocytes grown either in serum-free or in serum-containing medium. In both media, the addition of 10 microM forskolin from day 1 after cell plating to day 3 or 7 did not affect cell proliferation. Such stimulations also failed to enhance preadipocyte differentiation, as assessed by the measurement of lipoprotein lipase (LPL) and glycerol 3-phosphate dehydrogenase (GPDH) activities, two markers of adipose conversion. Similar results were obtained when various concentrations of forskolin (0.1 nM-100 microM) were added for 2 days either during the growth phase (days 1-3) or after confluence (days 5-7). Addition of methylisobutylxanthine (MIX) or 8-bromo-cAMP was also found inefficient to stimulate porcine preadipocytes differentiation clearly. By contrast, post confluence treatment of the murine 3T3-L1 cell line with either forskolin or MIX markedly enhanced lipid accumulation and led to a dramatic increase in GPDH activity (up to 120 times). This indicates that similar culture conditions are adipogenic for the murine 3T3-L1 preadipocytes but not for porcine preadipose cells. In summary, this work clearly highlights the finding that porcine preadipocytes do not respond to classic activators of the cAMP pathway like rodent cells do. This calls in question again the general model proposed for the action of this pathway in adipose conversion and suggests that the mechanisms regulating adipocyte differentiation may differ among species. PMID- 10374263 TI - Defects of basement membrane and hemidesmosome structure correlate with malignant phenotype and stromal interactions in HaCaT-Ras xenografts. AB - Benign and malignant HaCaT-ras clones, derived from immortalized HaCaT cells were grown as nude mouse surface transplants rendering a human tumor progression model. Searching for malignancy-related alterations, the deposition, localization and mRNA of basement membrane and hemidesmosome components were analysed by immunofluorescence, in situ hybridization and electron microscopy. Initially, at 1 week epithelia of benign and malignant cells revealed a similarly low polarity and an enlarged 'activated basal' compartment, reflected by partial dislocation and extended pericellular staining of the hemidesmosome constituent integrin alpha 6 beta 4 seen by immunofluorescence. Whereas benign grafts eventually normalized, closely resembling grafts of HaCaT cells, malignant growth was correlated with a persisting epithelial activation state and continuing higher expression of alpha 6 (by immunofluorescence and in situ hybridization). The basement membrane components bullous pemphigoid antigen 1, laminin-5 and collagen IV exhibited a largely linear distribution at 1 week. However, in the malignant cell transplants initially minor basement membrane discontinuities became more severe at around 2 weeks, associated with close stromal cell contacts, angiogenesis and invasion. Most striking were basement membrane alterations seen by electron microscopy. At 1 week stretches of basement membrane had developed in malignant transplants, though to a much lesser extent than in benign specimens. With invasion these basement membrane structures mostly disappeared despite persistent although variable immunofluorescence, suggesting high turnover without ultrastructural assembly. The hemidesmosome structures were defective throughout, completely lacking anchoring plaques with keratin filaments, whereas they were still associated with basement membrane deposits. Thus, malignant HaCaT-ras transplants, while initially resembling regenerating wounds, revealed an increasing loss of tissue polarity and basement membrane structures, which seemed to be accelerated upon stromal cell contacts. PMID- 10374264 TI - Desmosomal plakophilin 2 as a differentiation marker in normal and malignant tissues. AB - Plakophilin 2 (PKP2) is a widespread protein which shows a remarkable dual location: On the one hand, it appears as a constitutive karyoplasmic protein and on the other it is a desmosomal plaque component of most, probably all, desmosome possessing tissues and cell culture lines. Here we report on its desmosomal occurrence as revealed by immunocytochemical results obtained with three PKP2 specific murine monoclonal antibodies (mAbs) PP2-62, PP2-86 and PP2-150. These mAbs detect PKP2 in characteristic desmosomes of most normal cells, including simple and stratified epithelia as well as non-epithelial tissues such as myocardium and lymph node follicles. In addition, however, several normal tissues consistently display a differentiation-related PKP2 distribution, for example an absence of immunostaining in the "keratinizing" local specializations of the thymic epithelial reticulum, i.e. Hassall's corpuscles, and the restriction of PKP2 to the stratum basale of most stratified squamous epithelia, in contrast to its absence in upper strata, which contain PKP1- or PKP3-rich desmosomes instead. Taking advantage of the reactivity of mAb PP2-150 with formalin-fixed, paraffin embedded material, a series of human carcinomas (n = 37) has also been analyzed. The results suggest that mAbs to PKP2 may serve as markers for the identification and characterization of carcinomas derived from--or corresponding to--simple or complex epithelia. Thus consistent PKP2 immunostaining has been observed in all 18 cases of adenocarcinomas tested, but more variable and heterogeneous staining has been noted in squamous cell carcinomas, depending on the specific tumor type. The potential value of such mAbs for cell typing in normal and embryonic tissues and for detecting cell subpopulations with different degrees of differentiation is discussed with respect to their possible application in tumor diagnosis. PMID- 10374265 TI - Plakophilin 3--a novel cell-type-specific desmosomal plaque protein. AB - Desomosomes are cell-cell adhesion structures of epithelia and some non epithelial tissues, such as heart muscle and the dendritic reticulum of lymph node follicles, which on their cytoplasmic side anchor intermediate filaments at the plasma membrane. Besides clusters of specific transmembrane glycoproteins of the cadherin family (desmogleins and desmocollins), they contain several desmosomal plaque proteins, such as desmoplakins, plakoglobin, and one or more plakophilins. Using recombinant DNA and immunological techniques, we have identified a novel desmosomal plaque protein that is closely related to plakophilins 1 and 2, both members of the "armadillo-repeat" multigene family, and have named it plakophilin 3 (PKP3). The product of the complete human cDNA defines a protein of 797 amino acids, with a calculated molecular weight of 87.081 kDa and an isoelectric point of pH 10.1. Northern blot analysis has shown that PKP3 mRNA has a size of approximately 2.9 kb and is detectable in the total RNA of cells of stratified and single-layered epithelia. With the help of specific poly- and monoclonal antibodies we have localized PKP3, by immunofluorescence or immunoelectron microscopy, to desmosomes of most simple and almost all stratified epithelia and cell lines derived therefrom, with the remarkable exception of hepatocytes and hepatocellular carcinoma cells. We have also determined the structure of the human PKP3 gene and compared it with that of plakophilin 1 (PKP1). Using fluorescence in situ hybridization, we have localized the human genes for the three known plakophilins to the chromosomes 1q32 (PKP1), 12p11 (PKP2) and 11p15 (PKP3). The similarities and differences of the diverse plakophilins are discussed. PMID- 10374266 TI - [Transfusion-associated graft-versus-host disease]. PMID- 10374267 TI - [Role of antiplatelet in the prevention and treatment of thrombotic diseases]. PMID- 10374268 TI - [Pharmacokinetics and efficacy of low-dose all-trans retinoic acid in the treatment of acute promyelocytic leukemia]. AB - A clinical trial was conducted in order to evaluate the therapeutic effect and side effects of low-dose all-trans retinoic acid (ATRA) in the treatment of acute promyelocytic leukemia (APL). First of all, we compared the pharmacokinetic features in normal individuals with a single oral dose of ATRA at 15 mg/m2 and 45 mg/m2, respectively. The results showed that maximal plasma concentration with oral ATRA at 15 mg/m2 was high enough (10(-6) mol/L) to induce APL cell differentiation. Based on these results, 27 cases of de novo APL patients were treated with oral ATRA at a dose of 15-20 mg/m2/day and 24 of the 26 (92%) evaluable cases achieved clinical complete remission (CR) after 13 to 67 dyas of ATRA treatment. No patient experienced retinoic acid syndrome (RAS) and DIC. The Cmax with a single dose of ATRA on day 1 of treatment and immediately after obtaining of CR with ATRA in three cases demonstrated similar values in one patient and an about 2 fold decrease in two patients. Moreover comparison with a relatively well-matched previous group of 20 APL patients treated with high-dose ATRA showed that low-dose ATRA is as effective as high-dose in treating APL patients and may provide the advantages of decrease of the degree of hyperleukocytosis and side effects. PMID- 10374269 TI - [A study on relationship between chemotherapy of acute leukemia and apoptosis in vivo]. AB - In order to investigate whether apoptosis occurs in vivo in patients with leukemia in different stage (before, during and after chemotherapy). We detect apoptosis of peripheral blood leukemia cells in 10 patients before, during and after chemotherapy with in situ TdT fluorescence measurement and DNA electrophoresis. The results showed that apoptotic cells were few (0-2.9%) before chemotherapy, increased obviously (11.4%-72.0%) during chemotherapy in patients with complete remission (CR) or partial remission (PR), but decreased (0-2.9%) after chemotherapy (10-14 days). Apoptotic cells were few (0-5.7%) during chemotherapy in four patients without remission. Therefore, only effective drug therapy could induce apoptosis. DNA ladder cannot be detected by agarose gel electrophoresis either before, during and after chemotherapy. So we considered that in situ TdT assay is possible to apoptosis in vivo in the early course of chemotherapy for immediate modification of chemotherapy, while electrophoretic analysis is not sensitive enough to detect apoptotic cells in vivo. PMID- 10374270 TI - [The clinical and biological significance of P-glycoprotein expression in acute nonlymphocytic leukemia]. AB - To evaluate the clinical and biological significance of P-glycoprotein (P-gp) expression in adult acute nonlymphocytic leukemia (ANLL), P-gp was detected in 169 patients including 152 previously untreated cases, 7 refractory cases, and 10 cases at remission by using monoclonal antibody UIC2 and flow cytometry. P-gp was expressed in 29% of previously untreated cases, being less than in 71% of the refractory cases. P-gp expression was not found in patients at remission. Morphologically, P-gp expression was high in hybrid acute leukemia (67%) and acute monoblastic leukemia (47%) subtypes. Immunologically, P-gp expression was significantly associated with CD34, CD7, CD14 or CD42b/CD61. Cytogenetically, P gp expression was highly associated with poor prognosis abnormalities (54%), which was significantly higher than 7% of P-gp expression in good prognosis abnormalities. 23% of P-gp positive previously untreated ANLL (not including M3) achieved complete remission; this was significantly lower than 76% in P-gp negative cases. These suggested that P-gp expression is an index of poor prognosis in adult ANLL and P-gp positive ANLL has unique clinical and biological features. PMID- 10374271 TI - [HLA-DQA1 genes involved in genetic susceptibility to rheumatic fever and rheumatic heart disease in southern Hans]. AB - The incidence of rheumatic fever (RF) or rheumatic heart disease (RHD) is high in southern China. We studied the genetic susceptibility of HLA-DQA1 alleles to RF or RHD with emphasis on the mechanisms 106 unrelated healthy individuals and 54 patients with RF or RHD of Chinese Han nationality in Guangdong were included. DNA extraction from various biological material using phenol/chloroform method and HLA-DQA1 genotyping by PCR-PAGE and then with silver dyeing was used to show the electrophoretic patterns. A total 6 alleles of HLA-DQA1 were found. Increased allele frequencies of DQA1*0101 (31.48%, RR = 2.89, P < 0.005, EF = 0.206) and decreased allele frequencies of DQA1*0102 (1.85%, RR = 0.106, P < 0.005, PF = 0.134) were observed. Two increased genotyping of HLA-DQA1 (DQA1*0101/0301, chi 2 = 8.84, P < 0.005 and DQA1*0101/0401, chi 2 = 6.23, P < 0.0025) and decreased genotyping of DQA1*0102/0301 (chi 2 = 11.98, P < 0.005) were also observed. These findings suggested that DQA1*0101 contribute to genetic susceptibility for RF or RHD in Guangdong hans while DQA1*0102 to its genetic resistance. Digesting the genotyping of HLA-DQA1 may provide scientific basis for finding susceptible individuals to RF or RHD. Using PCR-PAGE and silver dyeing technique, a new genotyping method for HLA-DQA1, which is simple sensitive and precise, was established and applied. PMID- 10374272 TI - [Prevention and treatment of stroke after hypertension for ten years in Hunan Province]. AB - In this single blind, randomized multiple center clinical trial, the patients care period was divided into two stages. In the first stage (1985, 1-1989, 12), 984 cases were observed for 5 years whereas in the second stage (1990, 1-1994, 12), 2080 cases were observed and were subdivided into two groups, treat group and control, both were followed up for 5 years. The results showed that the prevalence of hypertension increased gradually. In the second stage, the average of blood pressure of the treatment group, using nitrendipine, was lower than that of the control group. There was significant difference between the two groups (P < 0.05). In the treatment group, the stroke and death rates were also decreased significantly as compared with the control group and the first stage group (P < 0.01). PMID- 10374273 TI - [Changes of gastric pits in intestinal metaplasia of gastric mucosa]. AB - To analyze the changes of gastric pits in intestinal metaplastic mucosa in 20 specimens of subtotal gastrorectomy and to evaluate the relationship between intestinal metaplasia and gastric pits. We made stereomicroscopic observation of the resected parts of the stomach by using dye-staining method. The width of gastric pits was measured by microscope and the histopathological examination was carried out. The features were divided into two groups: intestinal metaplasia and simple chronic superficial gastritis. The stereomicroscopic features of gastric pits were observed in 15.38% type BC, 28.21% type C, 25.64% type CD and 30.77% type D. The width of gastric pits was significantly different between intestinal metaplasia and simple superficial gastritis, especially obvious in severe intestinal metaplasia. The figures of gastric pits were mostly those a type C and type D and widened gastric pits were more obvious in intestinal metaplastic mucosa than in simple chronic superficial gastritis. PMID- 10374275 TI - [Clinical analysis in seventy-two patients with chronic thromboembolic pulmonary hypertension]. AB - To improve the identification of chronic thromboembolic pulmonary hypertension. We restrospectively analysed clinical data of 72 patients with chronic thromboembolic pulmonary hypertension (CTEPH). The levels of arterial blood gases appeared to be decrease in PaO2 and PaCO2, and increase in P(A-a)O2. There were 80.3% of unsymmetrical pulmonary hyperlucencies on chest radiograph and 76.4% of right ventricular hypertrophy on electrocardiograph, 92.5% and 97.1% of right atrial and right ventricular enlargement, respectively. 11.1% of pericardial effusion was noted on echocardiograph, and 98.5% of rise in pulmonary arterial pressure calculated by Doppler. The mean pulmonary arterial pressure was 6.50 +/- 1.80 kPa (48.75 +/- 13.50 mmHg), in part, by right cardiac catheterazation. Both the incidences of pulmonary embolism were 100.0% on pulmonary angiography and on radionuclide lung perfusion scan. There were 43.1% of the history of deep venous disorders and 75.0% of the positive findings by radionuclide venography in the lower extremities, respectively. The misdiagnostic and the lost diagnosis rate of prehospitalization accounted for 90.3%. The understanding of clinical manifestations and laboratory findings of CTEPH is important to promote diagnostic sense and level of CTEPH. PMID- 10374274 TI - [Fluconazole versus ketoconazole in systemic fungal infection: a double-blind randomized study]. AB - We carried out a multiple-center study, including a double-blined randomized clinical trial on fluconazole (Flu, group A) and ketoconazole (Keto, group B) and an open trial on Flu only for evaluating the efficacy and safety of Flu in treating deep and shallow fungal infection. 222 patients participated in the study and most of them had severe underlying diseases. The dosage and therapeutic duration were as follows: Flu 200-400 mg daily for 1-8 weeks in 34 patients with fungal infection and 150 mg as a single dose in 30 patients with fungal vaginitis; Keto 400 mg daily for 1-8 weeks in 30 patients with fungal diseases and for 5 days in 30 patients with fungal vaginitis. In the trial 124 patients were randomized to receive either Flu (64 patients) or Keto (60). The cure rate in group A and B was 81.3% (52/64) and 58.0% (35/60) respectively (P < 0.05). The fungal eradication rates of the two groups were 85.7% and 70.0% (P > 0.05) respectively. 98 patients entered the open trial. They included fungal infection of the respiratory tract and urinary tract, cryptococal meningitis, fungal sepesis and systemic dissemination of mycosis and fungal vaginitis. 84 (85.7%) were cured. The fungal eradication rate of this group was 92.9%. The side-effect rates of the three groups were 3.1% (2/64), 5.0% (3/60) and 6.1% (6/98) respectively. They were mild and transient vomiting, nausea, and anorexia. In group B and the open group however, there was one case of ALT elevation in each group (P > 0.05). This study suggests that Flu is a safe, effective and useful drug for the treatment of severe fungal infection. PMID- 10374276 TI - [Acyclovir and ganciclovir in the treatment of malignant hematological diseases]. PMID- 10374277 TI - [Abnormalities of hematopoietic stem cells in autoimmune diseases]. PMID- 10374278 TI - [Gastric epithelial proliferation and apoptosis in Helicobacter pylori infection]. PMID- 10374279 TI - [Intensifying the study of the proper treatment of digestive ulceration]. PMID- 10374280 TI - [Crohn's disease and Mycobacterium paratuberculosis]. PMID- 10374281 TI - [Transduction of the IL-2 gene into human gastric cancer cell: an experimental study]. AB - We evaluated the possibility of inducing a productive transfer of the IL-2 gene into human gastic cancer cells (GCC) and assessed the phenotypic and proliferative changes generated in the nude mice. The plasmid vector (BMGNeo-IL 2) carrying the human IL-2 gene was used to transduce the SGC-7901 GCC line by the lipofectin reagent. The IL-2 gene was analyzed by Southern blot and productive IL-2 release using IL-2-dependent CTLL. Cytotoxicity of LAK cells was tested by MTT colorimetry. The kinetics of in vitro growth and proliferation of parental and engineered cells were also measured. Parental and IL-2 gene transduced GCC were injected into nude mice. The tumorigenic potential of IL-2 gene-transfected GCC was evaluated by examining their in vivo growth in nude mice. The productive insertion of the IL-2 gene was achieved in SGC-7901. The amounts of IL-2 constitutively released by the engineered neoplastic cells ranged from 20 to 131 U/ml of IL-2 produced from 10(6) cells in 24 hours. Transduction of GCC with IL-2 gene did not modify the morphology and growth rate. IL-2 gene transfected cells demonstrated increased susceptibility to cell killing by LAK cells. IL-2 producing cells lost their tumorigenicity as evidenced by failure to grow in nude mice. The results demonstrate that IL-2 gene can be productively transduced into human gastric cancer cells without modifying their morphology and growth rate and this transduction leads to reduced or abrogated in vivo tumorigenic potential. PMID- 10374282 TI - [Mycobacterium paratuberculosis in the intestine of patients with Crohn's disease]. AB - Crohn's disease has long been suspected of having a mycobacteria, specifically M. paratuberculosis cause. In this study polymerase chain reaction (PCR) assay was used to detect M. paratuberculosis DNA in 74 paraffin wax embedded gut tissues from endoscopic biopsy or surgical resection specimens, including 36 Crohn's disease tissues, 18 ulcerative colitis tissues and 20 non-inflammatory bowel disease tissues. Oligonucleotides derived from IS900 sequence, which is a repeated in M. paratuberculosis chromosome and highly specific for the M. paratuberculosis, were used as primer. The specificity of products was confirmed by southern blot hybridization by using a biotin-labeled probe. The results show that M. paratuberculosis DNA was detected in 17 Crohn's disease (47.2%), 2 ulcerative colitis (11.1%) and 3 non-inflammatory bowel disease control subjects (15.0%). PCR positive or negative results in 36 Crohn's disease show that there is no relation with small or large gut involvement, and also the presence or absence of granulomata. The results suggest that M. paratuberculosis is surely present in tissues from some patients with Crohn's disease, which may have a specific association with Crohn's disease. PMID- 10374283 TI - [Mast cells of ileocecal junction in irritable bowel syndrome]. AB - To investigate whether the mast cells (MC) of the ileocecal junction (ICJ) is elevated in the irritable bowel syndrome (IBS) and the possible roles of the MC in IBS, the biopsies of ICJ were stained specifically by histochemistry for the MC in the IBS group (n = 20) and the normal group (n = 19). The structure relation between MC and nerves was studied through an electronic microscopy and an immunohistochemical method demonstrating neuronspecific enolase. The results demonstrated that the number of the MC of ICJ was significantly elevated in the IBS (P = 0.019) and that mast cells were close to nerves which were often unmyelinated nerves in lamina propria. The results indicate that the MC of ICJ may be responsible for the pathophysiology of the IBS. We conclude that the MC of ICJ may be a mediator between the gut and the nervous system in the IBS, and that the mast cell stabilizer or the antagonists of the mast cell products may have potential treatment effects on the IBS. PMID- 10374284 TI - [DNA typing for HLA-DR and HLA-DQ alleles in Chinese patients with rheumatoid arthritis]. AB - We analysed the associations of DR and DQ alleles with rheumatoid arthritis (RA) in Han nationality of China. PCR-RFLP technique was used to determine DRB1, DQA1 and DQB1 alleles in 35 unrelated patients with RA and 100 healthy controls from the Hans. The frequency of DR4 was 51.4% in RA patients and 24.0% in the healthy controls (P < 0.01, RR = 3.30). Subtyping of DR4 revealed no differences in the relative distributions of the DR4 alleles between the RA group and the controls, which both were predominated by DRB1 * 0405 allele, but there was a significant increase in the absolute frequency of DRB1 * 0405 in RA patients compared with the healthy controls (31.4% vs 12.0%, P < 0.01). DQ4 (DQB1 * 04) was also increased both in the RA patients compared with the healthy controls (37.1% vs 10.0%, P < 0.001) and in DR4+ RA patients compared with DR4+ healthy ones (72.2% vs 41.6%, P = 0.0376). Further analysis showed that the haplotype DR4-DQ4 was associated with RA (RR = 5.17, P < 0.001), and that the DR4(+)-DQ4+ patients had more severe disease than the DR4- patients. These results suggest that DR4, mainly the DRB1 * 0405, is associated with RA in Han nationality, and DQ4 may play a role in DR4 conferring susceptibility to RA. The DR4-DQ4 haplotype could be taken as a predictive marker for disease severity. PMID- 10374285 TI - [Detection of circulating autoantibodies to beta 2-adrenergic receptors in patients with asthma]. AB - To understand the roles of autoantibodies to beta 2 adrenergic receptors in the pathogenesis of asthma, we investigated the positive chronotropic action of beta 2-selective adrenergic agonist, clenbuterol, on cultured neonatal rat cardiomyocytes. Thereafter, we detected the autoantibodies to beta 2-adrenergic receptors in the sera from patients with asthma as it could inhibit the positive chronotropic action of clenbuterol. In the sera of all patients with asthma (16 cases) there were autoantibodies to beta 2-adrenergic receptors, but not in the normal controls (20 cases). Further study showed that the inhibitory autoantibodies were IgG type. This experiment suggests that the autoantibodies to beta 2-adrenergic receptors may play an important role in the pathogenesis of asthma. PMID- 10374286 TI - [Lipid-lowering effect of pravastatin on glomerulosclerosis protection and treatment]. AB - The lipid-lowering effect of pravastatin on glomerulosclerosis was observed in nephrotic syndrome model, which was induced in rats by using repeated puromycin injections. The histologic changes of the model were similar to those of human focal segmental glomerulosclerosis. Rats were divided into three groups, namely, normal control, nephropathy and nephropathy with pravastatin. Pravastatin was administered through gastric tube 6mg.kg-1.d-1 for 12 weeks. The lipoprotein profile and histologic development were observed. The results showed that pravastatin-treated rats had significantly lower cholesterol, triglycerides, very low density lipoprotein and high density lipoprotein than the nephropathy group at 7 weeks (P < 0.05). Moreover, the renal damage was also less marked in pravastatin-treated rats. Light microscopic and immunohistochemical examination revealed that lipid-lowering therapy was associated with significant lower level of glomerular sclerosing index and less accumulation of extracellular matrix including collagen III, IV, laminine and fibronectin as compared with the nephropathy group. However urinary protein and serum albumin levels were similar in both groups. It is suggested that lipid-lowering thyrapy may retard the progression of glomerulosclerosis and thus preserve renal function. The results imply that hyperlipoidemia plays an important role in the pathophysiological progression of renal disease. PMID- 10374287 TI - [Antiviral effect of antisense oligodeoxynucleotides complementary to hepatitis B virus X gene in vitro]. AB - It has been reported that the products of the human hepatitis B virus (HBV) X genes can transactivate a variety of viral and host promoters including the X, core, pre S2/S and pre S1 promoter. In order to investigate their antiviral effect in cell culture, three pieces of antisense phosphorothioate oligodeoxynucleotides (ASON) complementary to HBV X genes 1510-1530, 1555-1581, 1768-1791 regions were synthesized. The specificity of the inhibitory effect of the ASON was determined by using 2, 2, 15 cells by ELISA, PAP-ELISA and in situ hybridization to detect HBsAg, HBeAg, HBxAg and HBV DNA. The results showed that these ASON could inhibit the expression of HBxAg, as well as HBsAg and HBeAg, with the inhibitary rates of 78.07%, 80.65% and 62.76% respectively. In situ hybridization results indicated that HBV DNA replication can also be inhibited. The decrease in virus production and the ammount of HBV DNA were dose and time dependent. The mechanism of action may be due to the inhibition of HBxAg by sequences specific to ASON, and then the decrease of its transactivating effect for HBV DNA promoter. PMID- 10374289 TI - [Advances in the drug therapy of rheumatoid arthritis]. PMID- 10374288 TI - [The role of insulin resistance in the clustering of cardiovascular risk factors]. AB - The coexistence of multiple cardiovascular risk factors in the same individual is called a clustering of cardiovascular risk factors. Many studies have shown that this phenomenoa is associated with a increased morbility of cardiovascular disease, and insulin resistance may be the common link between the risk factors. This study was designed to link the clustering of cardiovascular risk factors with insulin resistance. Using a new insulin sensitivity index [-log (fasting serum Glu x Ins)] to evaluate the insulin resistance, we investigated the relationship between the insulin sensitivity and cardiovascular risk factors in 106 patients with hypertension or coronary heart disease and 32 normal subjects. The normal group without cardiovascular risk factors was selected as controls. The individual with 2 risk factors were defined as having mild clustering, and those with > or = 3 risk factors as severe clustering. As the number of risk factors increased from 1 to > or = 3, the insulin sensitivity index increased from -1.69 +/- 0.24, -1.95 +/- 0.17 to -2.14 +/- 0.21 respectively. The insulin resistance was positively correlated with serum uric acid in women (P < 0.05), but not in men (P > 0.05). The insulin sensitivity was more decreased in postmenopausal than in pre-menopausal women. The insulin sensitivity index was 2.23 +/- 0.39 vs -1.73 +/- 0.13 (P < 0.05). The insulin resistance is associated with a clustering of risk factors of cardiovascular disease, more cluste-ring, more resistant. PMID- 10374290 TI - [Relation between Helicobacter pylori and stomach cancers]. PMID- 10374291 TI - [Application of image materials for diagnosis of nervous system diseases]. PMID- 10374292 TI - [Advantages and disadvantages of thrombolytic therapy of unstable angina pectoris]. PMID- 10374293 TI - [Systemic manifestations of myasthenia gravis and its putative pathogenesis]. AB - We studied the systemic manifestations of myasthenia gravis and its putative pathogenesis. 644 clinically, pharmacologically and electrophysiologically diagnosed patients with myasthenia gravis (MG) were studied. 33 patients had Graves disease, 5 periodic paralysis, 4 Hashimoto disease, and 4 epilepsy, polymyositis, rheumatoid arthritis and Guillain-Barre syndrome each. 45 patients had positive anti-skeletal-muscle antibodies, 9 positive anti-thyroid antibody, and 3 positive SSA and SSB. 11 MG patients with positive pyramidal signs (MG+PS) had no evidence of multiple sclerosis (MS) or other neurologic diseases. Compared with 12 MG patients without pyramidal signs (MG-PS) and 23 normal controls (NCs), they had not only high levels of IgGcsf and IgG syn, but also increased ratio of AChRAbcsf/AChRAbs (P < 0.05). Four MG patients had epilepsy. Four MG patients had severe memory disorders. Three MG patients had diseases of the peripheral nerves without diabetes mellitus and toxin. SGPT was significantly increased in 22 MG patients than in 9 normal controls (P < 0.05). These systemic manifestations were improved while MG was improved by immunological therapy. MG is a systemic autoimmune disease predominantly involving AChR on the postsynaptic membrane of N M conjunction. PMID- 10374294 TI - [Intelligence, memory and event-related potentials in patients with multiple sclerosis]. AB - To learn the characteristics of cognitive disturbance of patients with multiple sclerosis (MS). We divided 55 patients with MS into two groups and 71 normal subjects were tested with the Wechsler Adult Intelligence Scale, Clinical Memory Scale and Eventrelated potentials (ERP). Disturbances of intelligence and memory occurred along the course of MS at various degrees. The memory quotient (MQ), full intelligence quotient (FIQ), verbal quotient (VIQ) and proce-ssing quotient (PIQ) of patients with the intelligence quotient (IQ) less than 80 scores were 76%, 32%, 24% and 70% respectively (P < 0.01). The scores of MQ, FIQ and VIQ in patients with MS of cerebral type or cerebro-spine type was lower than those of the spine type (P < 0.005). The score of patients with MS in active stage was less than that of MS in inactive stage (P < 0.001). The intelligence disturbances of MS patients were usually characterized by subcortical dementia and showed slight or moderate degree by DSM-III criteria. In patients of chronic active type, it may be characterized either by cortical dementia or subcortical dementia and showed moderate or severe degree. MS2 patient had significantly prolonged N2 and P300 latencies difference as well as low P300 amplitude compared with controls (P < 0.001). The disterbance of intelligence and memory with MS shows the important implication for the course of MS and the therapeutic effect as well as the prognosis. PMID- 10374295 TI - [Relationship between enteroviral infection and dilated cardiomyopathy]. AB - To investigate the role of enteroviral infection in the patho genesis of dilated cardiomyopathy (DCM), we used the nested reverse transcription polymerase chain reaction to detect enteroviral RNA in 21 myocardial specimens and 17 peripheral blood samples from patients with DCM and 21 patients (controls) with cerebral trauma. Enteroviral RNA was detected in 9 of 21 myocardial tissues and 6 of 17 blood samples, whereas all controls myocardial samples were negative. In addition, antibodies of coxsackievirus group B were detected in 4 of the 17 patients with DCM by neutralization test. The present study suggested that DCM might be associated with persistent enteroviral infection. PMID- 10374296 TI - [Experimental studies about effect of dexamethasone on diaphragm of normal rats]. AB - The influence of dexamethasone on diaphragm function, its oxidative capacity, fiber cross-sectional areas, the content of glycogen and ultrastructural changes were determined in Wistar rats by receiving dexamethasone (2 mg/kg) muscle injection for two weeks. We noted dexamethasone treatment leads to significant atrophy of the mass of diaphragm, reduction of fiber cross-sectional areas (CSA) of type II a, II b fibers. Under high frequency (100 Hz) stimulation, the tention of the diaphragm muscle strips was decreased and diaphragm fatigue resistance (1/2 RT) was significantly improved. Histochemically, ATPase activity in type I and type II b fibers of the diaphragm was significantly reduced and a significant reduction of succinate dehydrogenase activity in the diaphragm was also observed, but increased glycogen was seen in the diaphragm. Ultrastructural changes including mitochondrial hyperplasia, swelling, myofibillae focus destructure were evident. It is concluded that steroid-induced myopathy may also involve the diaphragm. PMID- 10374297 TI - [Prevalence of diabetes and its risk factors in China 1994. National Diabetes Prevention and Control Cooperative Group]. AB - We studied the prevalence of diabetes and impaired glucose tolerance (IGT) and their risk factors in the population of China. It was a population-based, cross sectional study of 224,251 residents aged 25 years and over in 19 provinces and areas, including cities and rural areas of the North, South, East, West and central China. Using 1985 WHO criteria, We found the prevalence of diabetes and IGT were 2.51% and 3.20% respectively in 213,515 subjects aged 25 to 64 years. 70.33% of the subjects had newly recognised diabetes. The prevalence of diabetes in China is about 3 times higher than it was ten years ago, and the rate is increasing faster in the countryside than in cities. On average, subjects with diabetes are older, have higher personal annual incomes, and have more frequently a family history of diabetes. They also have higher mean body mass index (BMI), ratio of circumference of the waist to hip, systolic blood pressure, diastolic blood pressure and a greater prevalence of hypertension. They perform less physical activity and receive less education than persons with normal OGTT. Multiple logistic, stepwise regression analysis shows that age, BMI (or WHR), family history of diabetes, hypertension, less physical activity and higher annual income are independent risk factors of NIDDM, and that low education is also an independent risk factor of NIDDM in people with higher personal annual income. PMID- 10374298 TI - [Distribution of fibronectin in kidney of IgA nephropathy and its clinicopathological correlation]. AB - To understand of fibronectin (Fn) distribution in kidney of IgA nephropathy (IgAN) and their meaning, we studied 107 cases of IgAN. Strong positive staining of Fn was shown in mesangial areas with proliferation, crescents, segmental sclerosis, and sclerosing glomeruli. 33 patients (group M) had Fn distributed only in mesangium, and 74 (group B) had Fn along the capillary walls as well as mesangium. Compared with those in group M, patients in group B showed more severe renal histological lesions (P < 0.01), crescent formation and stronger staining of IgG, C3 (P < 0.05) and IgA (P < 0.01) along the capillary walls. More severe changes of GBM were found in group B under electron microscopy. Clinically, 24th protein excretion (P < 0.01), incidences of hypertension and nonselective proteinuria were significantly higher (0.01 < P < 0.05), and Ccr was lower (0.01 < P < 0.05) in group B. 44 patients were followed up for 19.2 months, and the incidence of renal function deterioration in group B increased much more (0.01 < P < 0.05) than that in group M. Our results dmonstrated that the increase of Fn along the golmerular capillary walls might reflect that excessive proliferation of mesangial matrix, severe immune injury in capillary wall and damage of GBM, which are correlated with unfavorable clinical features and progression of the disease. PMID- 10374299 TI - [The methylation of CT gene in chronic myeloid leukemia]. AB - To investigate the relationship between the abnormal methylation of the calcitonin (CT) gene and the karyotypic abnormality in predicting the disease progress in chronic myeloid leukemia (CML), we studied the abnormal methylation of the CT gene in 47 CML patients by using a sensitive PCR method. At the same time, cytogenetic study was made in the same group of patients. The abnormal methylation of CT gene was found in only 4 of 24 patients in chronic phase, but in 6 of 9 patients in accelerated phase, and in 12 of 14 patients in blast crisis. Sequential studies in one patient also showed that the gene is normally methylated during the chronic phase but turns hypermethylated as the disease progresses. Our findings indicate that abnormal methylation of the 5' region of the CT gene is regularly a marker of disease progression in CML. PMID- 10374300 TI - [Study on the relationship between primary Sjogren syndrome and HLA-DRbeta gene]. AB - The pathogenesis of primary Sjogren syndrome (PSS) is unclear yet. In order to investigate the role of genetic factor playing in the mechanism of this disorder, we studied the HLA-DR beta gene distributed in 70 patients with PSS and 136 normal subjects by using PCR-SSP technique. The results showed that the gene frequences of HLA-DR3, DR52 and DR2 in patients with PSS were significantly higher than that in normal controls. However, the gene frequencies of HLA-DR5 and DR9 in PSS patients were lower than that in the control group. There were the same genetic phenomena in the siblings of two PSS patients' families. We also found the relationship between HLA-DR52 and the anti-antibodies of SSA or SSB. Our results concluded that the genetic factor is involved in the pathogenesis of primary Sjogren syndrome. PMID- 10374301 TI - [Hepatitis C virus infection among long-term hemodialysis patients]. AB - We determined the prevalence and incidence of hepatitis C virus (HCV) infection among hemodialysis patients and evaluate their risk factors. 80 patients on maintenance hemodialysis in Peking Union Medical College Hospital Dialysis Center from December 1994 to June 1995 were studied. 30 medical students were used as health controls. Serum samples were tested for HCV antibodies by a second generation enzyme-linked immunosorbent assay (ELISA) and for HCV RNA using nested polymerase chain reaction (PCR) and retested for anti-HCV and HCV RNA three months later. 24 (30%) of 80 hemodialysis patients were anti-HCV positive, and 2 (3.6%) of 56 anti-HCV negative cases were found HCV RNA positive. By combined assessment, the HCV infection rate of hemodialysis patient was 32.5% (26/80). Mantel-Haenszel analysis showed that HCV infection was associated with multi transfusions, dialysis over a long period, renal transplantation, history of operation. Samples of dialysate in 9 serum HCV RNA positive cases were directly detected for HCV RNA, and 3 were found HCV RNA detectable. HCV infection rate in hemodialysis patients is higher than that of general population. The main risk factors for HCV transmission is transfusion of unscreened blood but is not an independent factor, hemodialysis itself plays an important role, the length of time on dialysis, renal transplantation, blood-contaminated material and contaminated dialysis equipment are also the risk factors. Infected hemodialysis patients should be isolated. PMID- 10374302 TI - [Antibacterial activity in vitro and clinical use of sulperazon]. AB - In order to approach antibacterial activity in vitro and the efficacy of Sulperazon (SPZ) (sulbactam/cefoperazone), the sensitivity tests of 1,372 strains from clinical isolated bacteria to 17 antibiotics including SPZ were determined. The Gram negative bacteria occupied 1,035 strains (75.4) and Gram positive 337 strains (24.6%). 50 episodes of infections of major respiratory system in 43 patients were treated by SPZ. 58% of bacterial infections occurred in hematologic malignant diseases and solid tumors patients. 24% of 50 episodes were in neutropenia status. The positive rate of bac-terial cluteres was 56% in the series. 1.0-2.0 g SPZ was administered twice a day for 5-18 days, 56% of them was more than seven days (28/50 episodes). The results of susceptibility tests showed that sensitive rates were most high and the nesistant rates of SPZ were lower than these of to common Gram negative and Gram positive bacteria in third generation cephalosporins. The efficacy rate of SPZ in clinical use was 84% (42/50 episodes), bacterial clearance rate was 89% (25/28 episodes). Three cases (6%) had temporary elevation and other adverse reactions of SPZ were not seen. PMID- 10374303 TI - [Left ventricular hypertrophy remodeling in hypertension]. PMID- 10374304 TI - [Impaired glucose tolerance: progress in research and challenge]. PMID- 10374305 TI - [Proper use of oral hypoglycemic agents]. PMID- 10374306 TI - [Latent autoimmune diabetes mellitus in adults]. PMID- 10374307 TI - [Clinical characteristics and main diagnostic points of latent autoimmune diabetes mellitus in adults]. AB - To understand latent autoimmune diabetes mellitus in adults (LADA), we compared the clinical characteristics, fasting plasma glucose and C-peptide level, genetic frequency of HLA-DQA1, -DQB1 chain in 25 patients with LADA, 57 patients with insulin-dependent diabetes mellitus (IDDM, 21 patients with children-onset IDDM, 36 patients with adult-onset IDDM with ketosis), 38 patients with NIDDM (mild and moderate 30 patients and severe 8) and 42 normal persons. The onset of age was 20 48 years old associated with obvious polyphagia, and weight loss. Body mass index (BMI) was < or = 25 and fasting plasma glucose was > or = 16.5 mmol/L (297 mg/dl). Fasting and 1, 2 hour post prandial C-peptide level showed low and flatter curve (0.4, 0.8 and 0.8 nmol/L respectively). Glutamate decarboxylase (GAD) antibody was positive. HLA-DQ beta chain substitution of aspartate molecule was at position 57 (susceptic gene). LADA could be diagnosed if a patient has the first point and any point of the second to the fourth point. Patients with LADA should take diet, exercises, especially insulin as early as possible in order to control fasting and post prandial plasma glucose, and prevent from further destroy of residue islet B cells and reduce diabetic complications of eye, kidney and nerve. PMID- 10374308 TI - [Clinical significance of microtransferrinuria in diabetic patients]. AB - We studied the relationship between the urinary microtransferrin (TRF) and early glomerular damage in diabetes mellitus. 61 patients with non-insulin-dependent diabetes mellitus and 40 healthy subjects were measured for urinary TRF with rate immunonephelometry assay. The Urinary TRF concentrations were found to be greatly elevated in patients with diabetes compared with those in the health subjects (P < 0.001). The increase of TRF was closely related to age and duration of diabetes mellitus, blood glucose, hypertension, retinopathy, and it was initial parameter to predict diabetic nephropathy. There was a significant correlation among urinary TRF concentration, microalbumin, and N-acetyl-beta-D-glucosaminidase. The urinary excretion of TRF was more elevated than that of microalbumin. It is suggested that excretion of TRF in urine is a more sensitive index of the glomerular dysfunction in diabetes mellitus. PMID- 10374310 TI - [Relationship between insulin resistance and coronary heart disease in non insulin-dependent diabetes mellitus patients]. AB - Recent clinical studies showed that an independent association between hyper insulinaemia and coronary heart disease (CHD) in men with normal glucose tolerance. In non-insulin-dependent diabetes mellitus (NIDDM) it is less clear. Therefore, we evaluated insulin-sensitivity index, plasma glucose, insulin, lipoproteins and apolipoproteins in 40 male NIDDM patients with CHD, and compared them with 36 male NIDDM patients without CHD. The insulin-sensitivity index determined by the reverse of fasting plasma glucose and insulin product. The results showed that the subjects with CHD had significantly lower insulin sensitivity index (P < 0.01) compared to those without CHD. Using step-wise logistic regression analgsis, we found insulin-sensitivity indexes (OR 0.237, 95%CI 0.0909-0.6167, P = 0.0032) were negatively associated with CHD, and were independent from other cardiovascular risk factors such as age, body mass index, hypertension, and hyperlipemia. We conclude that in NIDDM patients with CHD are more insulin resistant compared to those without CHD. Insulin resistance is associated with CHD, and independent from other cardiovascular risk factors. PMID- 10374309 TI - [A study on the possible mechanism for the second control site of insulin secretion in islets]. AB - Glucose stimulation of insulin release involves closure of ATP-sensitive K+ channels (K(+)-ATP channels) in beta cells. However, by using diazoxide to open K(+)-ATP channels, it has been reported that another mechanism exists, by which glucose can control insulin release independently from changes in K(+)-ATP channel activity. These data suggested that there is a second control site in islets when insulin is secreted. To probe the possible existence of this site, mouse islets were used to investigate the energy state of islets in insulin secretion when the activity of K(+)-ATP channels was eliminated by diazoxide. In the present study, isolated islets were cultured in PRMI 1640 medium for 18 hours. The islets were then divided into 6 groups randomly, with 10 islets per group, placed in 1 ml KREBS medium containing 30 mmol/L K(+) and 250 mmol/L diazoxide with 0, 3, 6, 10, 15, 20 mmol/L glucose respectively and incubated in 37 degrees C water for 60 minutes. A portion of the supernatant was withdrawn at the end of the incubation for insulin assay, the islets were treated with trichloracetic acid and diethyl ether for the measurements of ATP, ADP, GTP and UTP. The above findings demonstrated that insulin secretion increases with the rise of glucose concentration. In the same time, the level of ATP increaseded and the level of ADP decreased gradully. The ratio of ATP/ADP in islets correlated with the insulin secretion very well. The level of GTP and UTP also increased with the rise of glucose concentration. It is suggested that glucose can control insulin secretion independently from its action of K(+)-ATP channels and the existence of a second control site as an another mechanism for insulin release. The ratio of ATP/ADP is an important factor in this mechanism; GTP and UTP also participated in the control of insulin secretion. PMID- 10374311 TI - [Effect of somatostatin on human sphincter of Oddi motility]. AB - The effect of somatostatin on the sphincter of Oddi was investigated in 20 subjects referred for endoscopic sphincter of Oddi manometry. Six patients had common bile duct dilatation, three had common bile duct slight dilatation, three had hepatic hilar carcinoma and eight had normal findings on endoscopic retrograde cholangiopancreatography. A triple-lumen low compliance system was used to record the sphincter of Oddi basal pressure, phasic contraction frequency, amplitude, duration, and direction of wave propagation before and after intravenous administration of somatostatin in a dose of 250 micrograms. After a mean latency period of 1 min, there were significant changes including decreased basal pressure (P < 0.01) and decreased frequency of wave contraction (P < 0.01). The amplitude and duration of wave contraction and wave propagation sequence were not significantly influenced. Thus somatostatin has a significant inhibitory effect on the sphincter of Oddi activity, which may be helpful for biliary and pancreatic flow. PMID- 10374313 TI - [Epirubicin containing regimens in advanced malignant tumors report of 516 cases. Epirubicin Collaborative Study Group]. AB - In a prospective multi-centre collaborative study, 516 patients with advanced cancer were treated by epirubicin (pararubicin, EPI) containing regimens. After CEOP (cyclophosphamide CTX, EPI, vincristine VCR and prednisone PDN) was used in the treatment of 213 patients with non-Hodgkin's lymphomas, 87 patients had complete remission (CR) and 99 partial remission (PR). Their response rate was 87.3%. However, there were 2 CR and 71 PR in 161 patients with non-small cell lung cancer treated by CEP regimen (CTX, EPI and cisplatin PDD), with a response rate of 45.3%. In 70 breast cancer patients treated by EMF regimen (EPI, Methotrexate MTX and 5-fluorouracil 5-FU), 8 had CR and 28 PR, with a response rate of 51.4%. The EPI containing regimens were also effective in dealing with gastro-intestinal tract and nasopharyngeal cancers. Adverse effects of epirubicin containing regimens were mainly nausea and vommiting, and the dose-qlimit toxicity was leucopenia. Hepatic, cardiac and renal toxicities were rather mild. The current phase III study revealed that the effect of epirubicin is similar to that of adriamycin, but the cardiac toxicity is relatively mild. So the effects can be improved by increasing the dose-intensity. PMID- 10374312 TI - [Relationship between obese gene expressive product and obesity]. AB - In order to study the relationship between obese protein and obesity. A radioimmunoassay for human plasma obese protein (OP) was established using human OP (57-92) and its specific antiserum. We observed the distribution and the characteristics. Of OP in normal human and mouse tissue and changes of plasma OP in obese patients. We also observed the tissue distribution after intravenous injection of 125I-labeled OP fragments (116-167) and (93-105) in rats. The results were as follows. The contents of OP were much higher in brain tissue than those in abdominal adipose tissue; levels of OP in liver and in skeletal muscle tissue were zero; the contents of abdominal adipose tissue were higher in female mice than those in male mice; the contents of plasma OP in normal human beings were 194.3 +/- 17.7 ng/L, while those in mice were 2257.8 +/- 171.9 ng/L. It was also found that the administered OP fragments were widely distributed in various tissue and organs including the brain. Kidney was the richest in the OP fragments; liver and lung ranked second. The half time of the OP fragments in plasma clearance was about 4.5 min. The contents of OP in obese adults and in obese children were much lower than those in normal control group. There was significant negative correlation between OP levels and body mass index, serum concentration of glucose, cholesterol and triglycerides. So the present study indicates the important relationship between the changes of OP contents and the pathogenesis of obesity. PMID- 10374314 TI - [Efficacy of dihydroartemisinin in treatment of 37 malaria cases]. AB - Dihydroartemisinin is an efficacious derivative of artemisinin which can be taken orally in treatment of malaria with less side reactions. Thirty-seven cases with malaria were treated with this drug in Beijing and in Ruili City, Yunnan Province. Out of them, 25 cases suffered from falciparum malaria with an average parasitemia of 73,218/microliter and 12 from vivax malaria with an average parasitemia of 4,950/microliter. The dosage was 60 mg daily or 2 mg/kg for paediatric patients for 7 successive days with the first dose doubled. All the patients were clinically cured after treatment. Fever subsided 22-72 hours after the beginning of treatment in falciparum malaria patients with an average fever clearance time of 36.24 +/- 15.30 hours and their parasite clearance time was 24 72 hours with an average parasite clearance time of 44.80 +/- 19.09 hours. One of them had recrudescence 19 days after the disappearance of parasites, hence the recrudescence rate of falciparum malaria was 4.8%. Five early cerebral cases in this series were completely cured after the treatment. All the 12 cases with vivax malaria were also clinically cured after treatment, but 1 case had relapse 35 days after treatment. This patient was completely cured with another course of dihydroartemisinin combined with primaquine. All patients tolerated well the oral administration of dihydroartemisinin and no significant side reactions were seen. PMID- 10374315 TI - [Chromosomal abnormalities and their clinical significance in acute lymphoblastic leukemia in adults]. AB - To determine the chromosomal abnormalities and their clinical significance in adult acute lymphoblastic leukemia, karyotypic analyses with R- and/or G-banding were performed in 101 newly diagnosed ALL patients. The results showed that 69 (68.32%) cases had clonal chromosomal abnormalities and specific abnormalities including Philadelphia chromosome (32.67%), t(4;11) (2.97%), t(11;14) (3.96%), abnormalities of 8q24 (2.97%), 6q- (4.96%). Patients with Ph chromosome showed higher leukocyte counts and more frequent hepatosplenomegaly and most of them were L2 and B lineage subtypes. They had lower complete remission (CR) rate, needed longer time to achieve CR and relapsed earlier as compared with patients without Ph chromosome. These suggested that chromosomal abnormalities are very frequent and some specific abnormalities had specific clinical features. PMID- 10374317 TI - [Antiphospholipid antibodies and thrombosis]. PMID- 10374316 TI - [10-year epidemiological study on rheumatic diseases in Shantou area]. AB - 22,049 adults were surveyed in a ten-year epidemiological study. The frequency of common rheumatic symptoms in Shantou population was much lower than that in northern China. The prevalence of ankylosing spondylitis, rheumatoid arthritis, osteoarthritis, osteoporosis and gout was 0.20%-0.32%, 0.20%-0.26%, 8.3%-10.8%, 12.4% and 0.15%-0.17% respectively. The frequency of HLA-B27 was 4.1% among general population and 90.6% in ankylosing spondylitis. The most commonly involved sites of osteoarthritis were lumbar spine, neck, and knee; but hands and hip were rarely involved. 85% subjects under investigation were found to be short of calcium intake. The differences between the north and the south of China in the prevalence of rheumatic symptoms may be related to the diversities in consciousness of seeking medical advice, reduction of bone content, climate and ergonomics. PMID- 10374318 TI - [Study on p53, mdm-2 and p21WAF1 protein expression in ER-positive and ER negative human breast cancer cell lines and its relation to biological features]. AB - OBJECTIVE: To study p53, mdm-2 and p21WAF1 protein expression levels in ER positive and ER-negative human breast cancer cell lines and their relations to biological features. METHODS: Using cell culture, DNA stable transfection and immunohistochemical methods, the protein expression levels of p53, mdm-2 and WAF1 genes in ER-positive expressing wtp53 MCF-7 cells, ER-negative expressing mtp53 MDA-MB-231 cells and ER-transfected MDA-MB-231 cells were determined, and then, their relation to biological features were compared. RESULTS: (1) The function and expression of p53 protein of MCF-7 and MDA-MB-231 cells were obviously different. The expression levels of mdm-2 and p21WAF1 proteins in MCF-7 cells were higher than those of MDA-MB-231 cells (P < 0.05), The biological features of the former were more favorable than those of the latter. (2) The ER-transfected MDA-MB-231 cells showed lower expression of mtp53 and higher expression of mdm-2 protein (P < 0.05), but no significant difference from that of p21WAF1 protein (P > 0.05). Meanwhile, biological features leading to favorable prognosis were manifested. CONCLUSION: The ER status of breast cancer cell lines is related to the expression level of p53 and mdm-2 proteins and the biological features. PMID- 10374320 TI - [Study on measures to increase diagnostic accuracy of FNAC of breast masses]. AB - OBJECTIVE: To study the measures capable of increasing the diagnostic accuracy, reducing misdiagnosis as well as giving a full play to fine needle aspiration cytology(FNAC), in the diagnosis of breast masses, and to standardize the diagnostic report form. METHODS: Totally 1629 cases of FNAC were performed and 444 cases of them were checked with the histological diagnosis. RESULTS: The sensitivity of diagnosis of malignant tumors in 307 cases, the specificity of diagnosis of benign lesions in 137 cases and the overall accuracy of diagnosis were 95.8%, 98.5% and 96.6% respectively. The false negative rate, potential false positive rate and the overall misdiagnostic rate were 4.2%, 1.5% and 3.4% respectively, while no case of false positive diagnosis was found. CONCLUSIONS: We believe that: (1) It is very important to use a fine aspirator, to make smears of high quality, to carry out an integrated working procedure and to put into effect a conscientious check over cytological appearance with histological diagnosis. (2) The diagnositc report form introduced in "the uniform approach to breast fine needle aspiration biopsy" by Bethesda specialists is worthy of reference. (3) FNAC can further play important roles in the diagnosis and treatment of breast masses. PMID- 10374319 TI - [Study on the relationship between PS2 protein expression and prognosis in invasive breast carcinoma]. AB - OBJECTIVE: To study the relationship between the expression of PS2 protein and prognosis in the invasive breast cancer (IBC). METHOD: Using LSAB immunohistochemical method, PS2 protein expression in 86 cases of IBC was detected. RESULTS: The positive rate of PS2 protein was 66.27% (57/86) in 86 cases of IBC. Under the following 3 conditions, PS2 protein expression levels in the more-than-5-year-survival group (A) were higher than those in the less-than-5 year-survival group (B). (1) In 86 cases of IBC, the expression level was 80.55% (29/36) for Group A, Significantly different from the 56.00% (28/50) of Group B (P < 0.025). (2) In 62 cases of premenopausal patients, the corresponding data were 86.20% (25/29) and 54.54% (18/33) respectively, P < 0.005; whereas in 24 cases of postmenopausal patients, 4/7 and 58.82% (10/17) respectively, P > 0.5. (3) In 62 cases of axillary node positive patients, the expression levels for Groups A and B were 82.35% (14/17) and 55.55% (25/45) respectively, P < 0.05; while in 24 cases of axillary node negative patients, 78.94% (15/19) and (3/5) respectively, P > 0.5. CONCLUSIONS: These results suggest that the expression of PS2 protein was positively correlated with 5-year-survival in IBC, and could be considered as a prognostic predictor for breast cancer, PS2 protein expression was a useful indicator for endocrine therapy in premenopausal patients. In axillary node positive patients, the expression of PS2 protein was associated with a better prognosis. PMID- 10374321 TI - [The expression of proliferating cell nuclear antigen in breast phylloides cystosarcoma and its significance]. AB - OBJECTIVE: To investigate the expression of proliferating cell nuclear antigen (PCNA) in breast phylloides cystosarcoma and its clinicopathological signification. METHODS: Immunohistochemistry (SP method) with monoclonal antibodies PC10 against PCNA was performed in 100 cases of phylloides cystosarcoma and 39 cases of adenofibroma. RESULTS: The positive rate of PCNA in phylloides cystosarcoma was 86%. The average PCNA index (PI) of phylloides cystosarcoma was significantly different among every histologic grading (F = 85.33, P < 0.01). The degree of differentiation was lower, the PI was higher. The PI was closely correlated with histologic grading (r's = 0.77). The PI in grade I phylloides cystosarcoma was higher than that in the group of adenofibroma with abundant mesenchymal cells (t = 3.42, P < 0.01). There was only a low correlation between PI and mitotic figures in phylloides cystosarcoma (r = 0.39). CONCLUSIONS: These findings suggest that the detection of PCNA has considerable practical value in reflecting the proliferation of activity of phylloides cystosarcoma, assisting the pathologists to make histologic grading; distinguishing the malignancy from benign ones (differential diagnosis) and evaluating the prognosis. PMID- 10374322 TI - [Relationship between p16 and Rb protein expression in astrocytomas]. AB - OBJECTIVE: p16 and Rb protein were examined in primary astrocytomas in order to study the correlation between p16 and Rb proteins. METHODS: p16 and Rb pretoin were immunostained by SP immunohistochemical method in the sections of formalin fixed paraffin embedded tumor tissue from 102 patients with astrocytoma brain tumors. RESULTS: p16 and Rb protein were expressed in all low grade (WHO Grade I and II) astrocytomas but only in 48.1% and 57.4% astrocytomas of Grade III and Grade IV respectively, in which, 24/31 of Rb protein positive tumors showed null or low expression of p16 protein, while 19/23 of Rb negative tumors were associated with positive or high level expression of p16 protein. CONCLUSIONS: (a) p16 and Rb proteins were both involved in astrocytoma progression. (b) Negatively correlated pressionof p16 and Rb protein might be one of characteristics of malignant astrocytoma. PMID- 10374323 TI - [Expression of p53, c-erbB1, c-myc and p16 gene proteins in human glioma]. AB - OBJECTIVE: To realize the significance of c-erbB1 and c-myc protooncogenes, p53 and p16 tumor suppressor genes in the development and pathologic diagnosis of human glioma. METHODS: The expression of p53, c-erbB1, c-myc and p16 gene proteins was detected immunohistochemically in 65 cases of glioma. The relationship between these results and the pathological grades was determined. RESULTS: The overexpression of mutant p53, c-erbB1 and c-myc was in accordance with the increasing grade of glioma malignancy, showing significant difference (P < 0.05) between the benign group and highly malignant group in p53 and c-erbB1, while the deletion of p16 protein was more frequent in the highly malignant group than in any other group. These 4 genes showed co-expression in some of the gliomas. CONCLUSIONS: The 4 genes, especially p53 and c-erbB1, play an important role in the development of glioma; the detection of these gene proteins has a positive significance on malignancy determination for glioma. PMID- 10374324 TI - [Observation on oligodendroglioma with light, electron microscopy and immunohistochemistry]. AB - OBJECTIVE: To investigate the ultrastructural and histological characteristics of oligodendroglioma (ODG). METHODS: By means of light microscopy and partially electron microscopy and immunohistochemistry, 65 cases of ODG were observed. RESULTS: According to WHO classification of the tumor, all 65 cases of ODG were graded in the degree of differentiation of tumor cells, showing 19 cases of Grade I, 32 cases of Grade II and 14 cases of Grade III. CONCLUSIONS: (1) The presence of glial filaments in tumor cells may be explained as a heterogeneity in differentiation. (2) Besides the degree of differentiation, some other criteria such as cellular density and mitotic activity are also closely related to the grading of ODG. PMID- 10374325 TI - [Study on microsatellite instability in gastric cancer]. AB - OBJECTIVE: To evaluate the microsatellite instability (MIN) in gastric cancers (GC). METHODS: MIN was examined in 106 cases of GC, using the radioactive polymerase chain reaction-based method. RESULTS: The frequency of MIN was significantly higher in expanding type of GC as compared with the infiltrative type (P < 0.005, chi 2 test). No significant association with histologic type, pTNM stage, age and sex was found (P > 0.05, chi 2 test). CONCLUSIONS: These findings reveal that MIN might be an important contributor to the development and progress of a significant number of GC, especially the expanding type which, as acknowledged traditionally, has a favorable prognosis. PMID- 10374326 TI - [Observation on human embryonic lung fibroblast proliferation mediated by mitogen activated protein kinase]. AB - OBJECTIVE: To study the effect of mitogen activated protein kinase (MAPK) on the proliferation of human embryonic lung fibroblast (HELF) induced by 60Co irradiation and the relationship between MAPK and angiotensin II (A II). METHODS: Cell proliferation was measured with enzyme-labeling method; angiotensin II, MAPK and type I precollagen synthesis as well as the role of sodium nitroprusside on the inhibition of A II were evaluated adopting immunohistochemical technique combined with image analysis after 1-5 Gy 60Co gamma-ray irradiation on the cultured HELF. RESULTS: Irradiation at 1-5 Gy promoted cell proliferation and type I precollagen synthesis; the syntheses of A II and MAPK were also increased in the irradiated cells. Exogenous A II enhanced cell proliferation, but sodium nitroprusside inhibited the action of A II. CONCLUSIONS: HELF proliferation induced by 60Co irradiation is a chain reactive course, in which, A II and MAPK are involved. MAPK may play a role of limiting valve in the signal transduction of cellular proliferation. PMID- 10374327 TI - [Numerical change of 7, 17 and Y chromosomes in human lung carcinoma detected by fluorescence in situ hybridization]. AB - OBJECTIVE: To study the copy number of chromsomes 7,17 and Y in lung carcinomas. METHODS: Fluorescence in situ hybridization (FISH) was used to detect the number of above mentioned chromosomes on the touch smears of 17 cases of lung carcinoma (adenocarcinoma and squamous cell carcinoma). RESULTS: Triploid chromosome 7 FISH signals were found in 31.73% of the tumor nuclei, dipoid chromosome 17 FISH signals in 43.00% and monoploid chromosome Y FISH signals in 58.04% of these nuclei. CONCLUSIONS: The number of chromosome 7 was obviously increased in both lung adenocarcinoma and squamous cell carcinoma, and the rate of trisomy 7 was the highest. In addition, there is a significant difference in the distribution of chromosome 7 ploidy between lung adenocarcinoma and squamous cell carcinoma. PMID- 10374328 TI - [Overexpression of p53 protein and its relation to HPV in squamous intraepithelial lesion and cervical carcinoma]. AB - OBJECTIVE: To study the relation between the expression of p53 protein and HPV infection in squamous intraepithelial lesion (SIL) and cervical carcinoma. METHODS: The expression of p53 protein was determined in the paraffin embedded tissues of 171 cases of cervical carcinoma, 68 cases of SIL and 29 cases of chronic cervicitis, using ABC immunohistochemical method. Detection and typing of HPV were carried out adopting PCR-RFLP technique on the same tissues. RESULTS: In the cervical carcinomas, the overexpression rate (38.6%) and intensive overexpression rate (11.7%) of p53 protein were higher than those in LSIL (10.0% and 0.0%, respectively) and HSIL (35.4% and 2.1%, respectively). The overexpression rates of p53 protein in SIL and cervical carcinoma with positive HPV 18 were 87.5% and 84.3% respectively, higher than 21.2% and 10.5% in the corresponding groups carrying HPV 16, as well as 16.7% and 57.1% in the corresponding groups negative for HPV. CONCLUSIONS: The overexpression of p53 protein might play a role in the carcinogenesis of cervical carcinoma. The relation between p53 overexpression and HPV 18 infection in SIL and cervical carcinoma remains to be clarified. PMID- 10374329 TI - [Familial breast cancer-susceptibility genes]. PMID- 10374330 TI - [Pathological types and pathomorphological characteristics of osteosarcoma]. PMID- 10374331 TI - [Alterations of retinoblastoma gene and its protein expression in aggressive bone tumors]. AB - OBJECTIVE: To study the association between RB gene and the oncogenesis of bone tumors. METHODS: Southern blot and immunohistochemical techniques were used to detect the structural anomalies of RB gene in 34 cases of bone tumors and the expression of RB protein in 99 paraffin-embedded bone neoplasma. RESULTS: The deletion and/or rearrangement of RB gene were detected only in 42.9% (9/21) of osteosarcoma; lack of RB protein expression was noticed in 12 cases including 7 cases of osteosarcoma (7/26, 26.9%) and 5 cases of chondrosarcoma (5/23, 21.7%). Beniga giant cell tumor of bone and chondroblastoma showed positive RB protein expression; osteosarcoma cells presenting poor differentiation and apparent atypia all showed no expression of RB protein. Most of the high-grade chondrosarcoma also had no RB protein expression. CONCLUSION: The alterations of RB gene and loss of RB protein may play a role in the pathogenesis and progression of malignant bone neoplasms. PMID- 10374332 TI - [Correlation of tumor microvesseldensity with prognosis in osteogenic sarcoma]. AB - OBJECTIVE: To investigate the relationship between tumor angiogenesis and clinical pathology, as wel as the prognosis in osteosarcoma. METHODS: Intratumoral microvessel density (MVD) in 70 cases of osteosarcoma was assessed immunohistochemically by using the specific endothelial cell markers F VIII-RA and CD31. RESULTS: There was no correlation between the MVD and the tumor size, Price's grade, as well as Dahlin's classification; however, significant correlation was found between the MVD and the neoplastic osteoid classification, new WHO classification in 1993, as well as the proliferating cell nuclear antigen (PCNA) labelling index. Microvessel counts were associated with overall survival by Kaplan-Meier analysis. An average vessal count of less than 31 (x 200) suggested a better survival, but a higher vessel count of more than 31 (x 200) showed a trand to worse the overall survival. CONCLUSION: The results suggest a significant relationship between MVD and prognosis; moreover, MVD may be a useful prognostic indicator in osteosarcomas. PMID- 10374333 TI - [Alterations of MDM2 and p53 genes in bone tumors]. AB - OBJECTIVE: To determine the frequency of MDM2 and p53 genes and their possible role in the pathogenesis and development in bone tumors. METHODS: Digoxigenin labeling in situ hybridization technique was used to investigate the expression of MDM2 and p53 in 38 cases of bone tumors, including 12 osteosarcomas, 10 chondrosarcomas, 14 giant cell tumors of bone and 2 chondroblastomas. The relationship between MDM2 and p53 expressions was analyzed. RESULTS: The positive rates for MDM2 in osteosarcoma, chondrosarcoma and giant cell tumor of bone were 41.7%, 50.0% and 35.7%, while those for p53 were 58.3%, 40.0% and 21.4%, respectively. The two cases of chondroblastoma showed both MDM2 and p53 overexpression. There was a striking association between MDM2 and p53 overexpressions. CONCLUSION: MDM2 and p53 alterations are frequent events in bone tumors and may be involved in tumorigenesis or tumor progression in bone tumors. PMID- 10374334 TI - [Promotive effect of TNF-alpha and M-CSF on osteolysis induced by giant cell tumor of bone]. AB - OBJECTIVE: To explore the effect of cell components in giant cell tumor of bone (GCT) and cytokines expressed by them on osteolysis. METHODS: Mononuclear stromal cells and multinucleated giant cells (MGCs) were isolated from 10 cases of GCT, and fibroblast-like stromal cells were obtained by long-term culture of mononuclear stromal cells. Osteolytic capability of above isolated cells were tested in an in vitro cell-bone resorption model. RESULTS: All cell components isolated from GCT had capability to resorb bone matrix directly. Exogenous tumore necrosis factor-alpha (TNF-alpha) could significantly increase the bone resorption induced by both kinds of stromal cell. There were higher level of TNF alpha and greater expression rate of macrophage colony stimulating factor (M-CSF) in GCT than in the osteosarcoma tissues or the normal serum. CONCLUSIONS: The characteristic bone resorption behavior of GCT might be conducted by all it's three major cell components, and this bone resorption process could be promoted by both TNF-alpha and M-CSF they expressed. PMID- 10374335 TI - [Effects of excess fluoride on bone turnover under conditions of diet with different calcium contents]. AB - OBJECTIVE: To study the effects of excess fluoride on bone turnover under conditions of diet containing different amount of calcium. METHODS: The experiment was performed on rats raised on a balanced diet with adequate calcium or a monotonous diet with low calcium and given amount of fluoride in their drinking water (F, 100 mg/L) for 2 months or 1 year. RESULTS: Osteomalacia, osteoporosis and accelerated bone turnover were observed with elevated serum alkaline phosphatase (ALP), osteocalcin (BGP) and parathyroid hormone (PTH) in rats fed on low calcium diet and fluoridized water for 2 months. In the rats fed on adequate calcium diet and fluoridized water for 2 months, only slightly increased osteoblastic activity was found while the average width of trabecular bone was increased with elevated serum ALP activity in rats raised on the same diet and water for 1 year. CONCLUSIONS: The basic effect of excess fluoride on bone is the causation of a high bone turnover state which can also be induced to a milder extent by low calcium diet itself. Therefore, the formation of a high bone turnover state is the pathogenetic basis for low dietary calcium intake to exacerbate the severity of skeletal fluorosis. PMID- 10374336 TI - [The inhibitory effect of combined treatment of TNF alpha and antisense oligodeoxynucleotides on the growth of human pancreatic carcinoma cell line cells]. AB - OBJECTIVE: To study the inhibitory effect induced by combined treatment of TNF alpha and antisense oligodeoxynucleotide on the growth of human pancreatic adenocarcinoma cell line (PC-2) cells. METHOD: TNF alpha and synthesized antisense c-myc or Ki-ras phosphorothioate oligodeoxynucleotide (ASPODN) were added to the culture media of PC-2 cells, whereas the groups treated with TNF alpha or ASPODN alone were used as controls. The cell growth rate was estimated by cell count and MTT analysis, the endogenous target gene expression was studied by adopting RT-PCR-Southern blot technique, and cell apoptosis was detected in situ. RESULTS: The cell growth was inhibited much more obviously in the groups of combined treatment than in the groups treated with TNF alpha or ASPODN alone. Marked inhibition of endogenous Ki-ras and c-myc expression was observed in the groups treated with Ki-ras and c-myc ASPODN. The amounts of apoptotic cells were 8.0% and 8.4% in the groups of TNF alpha + Ki-ras ASPODN and TNF alpha + c-myc ASPODN, and were 5.2%, 4.8% and 5.4% for TNF alpha, Ki-ras ASPODN and c-myc ASPODN respectively. CONCLUSION: Our results demonstrate that the inhibitory effect induced by combined treatment of TNF alpha and ASPODN on cell growth was stronger than that induced by TNF alpha or ASPODN alone. PMID- 10374337 TI - [Effects of nordihydroguaiaretic acid on the growth and differentiation of SHG-44 glioma cell line]. AB - OBJECTIVE: To investigate the effects of nordihydroguaiaretic acid (NDGA) on the growth and differentiation of glioma cells and their mechanism. METHODS: A human glioblastoma cell line (SHG-44) was treated with NDGA in the medium or intracytoplasmic microinjection of NDGA. Changes of the growth, morphology, cell cycle and immunohistochemical features of the cell were investigated. RESULTS: It was found that the growth rates and proliferation activity were inhibited by 100 mumol/L NDGA in the medium, with cell cycles altered. NDGA could induce differentiation of these cells with the glial filaments increased, and the treated cells expressed more GFAP but less vimentin. The expression of p53 and bFGF were also decreased. Furthermore, the result of microinjection (1.5 x 10( 11) g/cell) showed similar but more rapid and permanent effects on the glioma cells. CONCLUSION: NDGA has multiple effects of growth inhibition, differentiation therapy, as well as angiosuppression on human glioblastoma cells. PMID- 10374338 TI - [Studies on gene expression of bcl-2 and its correlation with tumor cell apoptosis as well as expression of p16 in human neuroblastoma]. AB - OBJECTIVE: To elucidate the relationship between bcl-2 gene expression and the frequency of apoptosis of tumor cells, and study the expression of p16 tumor suppressor gene in neuroblastoma. METHODS: In situ hybridization and immunohistochemistry methods were used to study the frequencies of expression of bcl-2 and p16 genes in 20 cases of neuroblastoma. Meanwhile, an in situ apoptotic cell detection method was adopted to detect the apoptotic cells in these tumors, and the number of apoptotic cells was compared with the bcl-2 gene expression in each case. RESULTS: In situ hybridization revealed that the positive frequencies of both bcl-2 and p16 gene expression in 20 neuroblastoma specimens were 95%, and the expression rates at the protein level of these 2 gene products as detected by immunohistochemistry were both 100%. There was no significant difference between the positive rates obtained by these 2 methods. Comparing the bcl-2 expression and apoptotic cell number in these specimens, we found that the apoptotic cell number increased as the level of bcl-2 expression decreased. CONCLUSIONS: The bcl 2 gene was expressed in most of human neuroblastomas. The reverse correlation of bcl-2 expression and tumor cell apoptosis further confirms that bcl-2 as an important gene inhibiting cell apoptosis may indirectly promote the carcinogenesis of neuroblastoma. It seems that there was no significant loss of p16 gene expression in these cases. PMID- 10374340 TI - [Pathological features of hepatocellular carcinoma carrying hepatitis C virus antigen]. AB - OBJECTIVE: To analyze the pathological features of hepatocellular carcinomas (HCCs) carrying different hepatitis virus antigens histopathologically and systematically. METHODS: PAP and ABC kits were used in the immunohistochemical study, and CAS-200 System was applied in the image cytometry. RESULTS: As compared with HCCs carrying HBV marker(s), the HCCs carrying HCV marker showed more cases of clear cell type (7/9 vs 4/33), better differentiation of cancers, less necrosis of hepatocytes, milder lymphocyte infiltration in the hepatic sinuses or periportal areas (P < 0.01), higher incidence of bile ductule damages, and bore a close relation to the formation of lymphoid follicle in the surrounding tissues (P < 0.05). These patients were elder, with lower grade of symptoms and better prognosis after operation. CONCLUSION: HCCs carrying only HCAg have different pathological features and clinical characteristics from which carrying HBV marker(s). The results of image cytometry are in accordance with the biological behaviour of HCC. PMID- 10374339 TI - [Study on p53 protein expression, cell proliferation and apoptosis in benign and malignant brain tumors]. AB - OBJECTIVE: To explore the effect of p53 protein expression in brain tumors on the cell proliferation and apoptosis, and the relationship between above parameters and histological classification and malignant degree of these tumors. METHODS: The p53 protein expression, cell proliferation and apoptosis of 10 control brain tissues (CBT) and 80 brain tumor samples were studied by in situ cell death detection and immunohistochemistry. RESULTS: Sixty-nine of the brain tumors (86.3%) expressed p53 protein. The amount of tumor cells positively stained for p53 protein increased with the degree of malignancy. All the CBTs did not express p53 protein. The densities of cells positive for proliferating cell nuclear antigen and Ki-67 antigen staining in tumor groups were higher than that in CBT group and were elevated with the malignant degree and the level of p53 protein expression of the tumors. On the contrary, the density of apoptotic cell in tumor groups was lower than that in CBT group, reducing in accord with the increase of malignancy and the level of p53 protein expression of the tumors. CONCLUSIONS: These results suggest that above 4 parameters have a referential value for the evaluation of biological behavior of brain tumors. It is clear from the present study that the abnormality of expression and function of p53 protein correlates with the imbalance between cell proliferation and apoptosis. This may be one of the important factors in the development and progression of primary and metastatic brain tumors. PMID- 10374341 TI - [The effect of cationic ferritin to the permeability of renal glomerular basement membrane and the mesangial area]. AB - OBJECTIVE: To study the effect of cationic ferritin (CF) to the permeability of the glomerular basement membrane (GBM) and the mesangial area. METHODS: Single dose of CF 50mg/kg/rat given intravenously. Samples taken from the kidney were studied under electronic microscope by polyethylenimine (PEI) staining. RESULTS: The number of PEI granules in GBM changed obviously while those in the mesangial matrix (MM) remained unchanged. CONCLUSION: The negative charge in GBM serves as a charge barrier relating with proteinuria and the MM serves as the lymphatic pathway with no connection with proteinuria. PMID- 10374342 TI - [Prospect of the study on the relation between hepatitis C virus and hepatocellular carcinoma]. PMID- 10374343 TI - Induction of immune tolerance in adult rabbits undergoing heterotopic cardiac transplantation. AB - OBJECTIVE: To induce experimental immune tolerance in rabbits and observe its effects on heterotopic cardiac transplantation. METHODS: Donor's splenic lymphocytes pretreated with platinum metal chelator were injected into the recipient's mesenteric-portal vein. Cyclosporin A was perfused through the donor's heart. RESULTS: The injection of donor's splenic lymphocytes before transplantation could significantly prolong the survival time of the heterotopically transplanted heart. The effect of two injections was better than that of one. Radioactive tracer studies showed that the 99mTc-HMPAO tagged lymphocytes injected into the recipient rabbit were later concentrated in the liver, though initially they were distributed in multiple organs. The induced immune tolerance was antigen-specific, and it neither affect the other immune functions of the lymphatic system prominently nor exert any harmful effect on the recipient's liver and renal functions. The perfusion of cyclosporin A through the donor heart could block the glycosyl groups, such as D-glucose, D-mannose or N acetyl-galactosamine on the surface of the myocardial cells, thus might change the antigenic expression, effectively preventing rejection of the graft by the host, and might be considered as a new method to block graft rejection in cardiac transplantation. The combined use of the above-mentioned two methods acted on both the host and the donor, thus reducing the exposed antigens on the donor organ as well as the immune reaction against the donor antigens, and resulting in synergistic effect in inducing immune tolerance in adult rabbits, and resulting in relatively long-term survival of transplanted hearts. CONCLUSION: This report may provide the experimental basis for inducing immune tolerance in clinical transplantation. PMID- 10374344 TI - Clinical epidemiologic characteristics of 430 cases of gallbladder cancer. AB - OBJECTIVE: To make clear the incidence, clinical characteristics and possible regional difference of gallbladder cancer in China. METHODS: A total of 430 cases of gallbladder cancer from 28 hospitals between 1986-1996 were reviewed, according to a standard protocol called "the clinical epidemiological list of gallbladder cancer". RESULTS: The incidence of gallbladder cancer was higher in the females than in the males. There was significant difference in the incidence between the north and south of China, and between the mountain area and flatlands. Gallbladder cancer accounted for 1.6% of bile tract disease in the same period. Gallstones were found in about 50% of the cases of gallbladder cancer. The clinical symptoms included abdominal pain, ictus, etc. The major pathohistologic type was adenocarcinoma, and 58% of tumors were localized in the whole gallbladder. Metastasis occurred mainly along the biliary tract or directly to the bed of gallbladder and liver. Ultrasonography and CT were useful to diagnosis. The positive imaging diagnostic rate was higher in 1991-1996 than in 1986-1990 P < 0.05) [corrected]. The rate of operative resection was 100% for stage I and II disease, 75% for stage III and IV, and significantly lower for stage V (P < 0.05). The 3-year survival rate in patients with stage I or II disease was significantly higher than that in those with terminal cancer (P < 0.05). CONCLUSIONS: There is specific populational, time and regional difference in the distribution of gallbladder cancer. Ultrasonography and CT are the most important diagnostic methods. Early diagnosis and early radical resection are the key to increasing the 5-year survival rate. PMID- 10374346 TI - The effects of fibroblast growth factors on ischemic kidney, liver and gut injuries. AB - OBJECTIVE: To explore the possibility of reducing reperfusion injuries of internal organ with acidic and basic fibroblast growth factors (aFGF and bFGF). METHODS: Two kinds of ischemia and reperfusion animal models were used in this study. In rat model of superior mesenteric artery (SMA) occlusion, microvascular clamp was placed on the root of SMA to cut off the blood flow for 45 minutes, and then the clamp was removed. In rat model of bilateral renal ischemia and reperfusion, both renal arteries were clipped to get complete cessation of blood flow for 60 minutes, then the blood flow was allowed to return. At the onset of reperfusion, the doses of 4.0 micrograms/rat of bFGF in SMA occluded rats or 2.6 micrograms/rat of aFGF in rats with acute renal injury were administered through the jugular vein. The liver and renal function examination, tissue bacterial study and histopathological evaluation were done to evaluate the treatment results. RESULTS: The functional impairment of ischemic liver, gut and kidney were reduced with venous administration of aFGF or bFGF at the onset of reperfusion. The results of pathological and tissue bacterial examination supported the assertion of significant protective effects of FGFs. CONCLUSIONS: The protective effects of FGFs may come from the non-mitogenic effects of FGFs at the early and the mitogenic effects at the late stage of tissue repair. These results indicate a potential for clinical use of FGFs as a therapeutic modality in ischemic visceral organ injuries in the future. PMID- 10374345 TI - Microencapsulation of rat islets prolongs xenograft survival in diabetic mice. AB - OBJECTIVE: To protect the transplanted islets from the host's immune system by means of immunoexclusion membranes. METHODS: Rat islets were isolated from Wistar rat pancreas by ductal collagenase distention, stationary digestion, and finally with the aid of dextran gradient separation. Then the islets were encapsulated in alginate-polylysine-alginate (APA) semipermeable membranes. RESULTS: In vitro studies demonstrated that encapsulated islets secreted insulin in response to glucose challenge for at least 8 weeks, which was similar to free islets. In vivo studies showed that 15 streptozotocin (STZ)-induced diabetic mice were transplanted intraperitoneally with 1000 encapsulated islets without immunosuppression. Diabetes was reversed within 3 days, and the mice remained normoglycemic for up to 160 days, with a mean xenograft survival time of 126 days. The encapsulated islets had a significantly greater effect than unencapsulated islets, which functioned for less than 8 days. CONCLUSIONS: Encapsulation of pancreatic islets in semipermeable membranes can effectively prolong xenograft survival without immunosuppression in an animal model. PMID- 10374347 TI - Targeted diagnosis and treatment of superficial bladder cancer with monoclonal antibody BDI-1. AB - OBJECTIVE: To explore the application of monoclonal antibody (McAb) to targeted treatment of bladder carcinoma through a series of in vitro and in vivo studies carried out in animal model and patients with bladder carcinoma. METHODS: Monoclonal antibody BDI-1 against bladder carcinoma was prepared by the lymphocyte hybridoma technique. McAb was conjugated with 99mTc by direct reduction method. Momodin (MD) was covalently linked to McAb by SPDP method. Radioimmunoimaging of nude mice xenografts and patients with bladder carcinoma were performed with BDI-1-99mTc conjugates. An immunotoxin (BDI-1-MD) was inducted via a catheter into the bladder. Targeted treatment with BDI-1-MD was carried out in 18 patients. RESULTS: This study showed the specificity of McAb, and clear imaging of nude mice bearing xenografts. Distribution analysis of 99mTc BDI-1 in nude mice showed the highest value of T/NT in bladder tumor. Targeted diagnosis and treatment for patients by intravesical administration are very safe and effective. CONCLUSION: The bladder cancer seems an ideal model for diagnostic and therapeutic approaches using regional administration of McAb conjugates via a catheter direct into the bladder. PMID- 10374348 TI - Detection of Fas/APO-1 in six human urogenital malignant cell lines with flow cytometry. AB - OBJECTIVE: To investigate the expression of Fas/APO-1 in urogenital tumor cell lines. METHODS: With direct immunofluorescence, the expression of Fas/APO-1 in six urogenital malignant cell lines and one primary in vitro cultured normal renal fibroblast was detected by flow cytometry. RESULTS: Expression of Fas/APO-1 was detected in all six urogenital tumor cell lines, but with limited positive cell percentage and relatively lower fluorescence intensity, compared with expression of Fas/APO-1 in normal control of primary in vitro cultured renal fibroblast. CONCLUSIONS: Lower expression of Fas/APO-1 in urogenital malignant cell lines than that in normal cells might be the reason for occurrence and progression of urogenital malignant tumors. PMID- 10374349 TI - The effect of thyrotropin receptor antibodies on the proliferation of FRTL-5 cells and the expression of protooncogene c-fos mRNA. AB - OBJECTIVE: Hyperthyroidism and a diffuse goiter are the main symptoms of Graves' disease (GD) associated with autoantibodies to thyroid-stimulating hormone (TSH) receptor (TRAb). The present study was conducted to evaluate effects of autoantibodies in patients with GD (TRAb-IgG) on induction of the proliferation and c-fos mRNA expression in FRTL-5 cells (Fisher rat thyroid cell line). METHODS: Highly purified IgG fractions were isolated from 11 patients with GD, TRAb-IgG and 15 normal individuals (normal controls) with Protein A Sepharose CL 4B affinity column chromatograph. FRTL-5 cells, which had been grown to subconfluency and deprived of TSH for a few days. Then, these cells were used for measuring cAMP content, 3H-thymidine incorporation in cells and the expression of c-fos mRNA respectively. RESULTS: After stimulation of TRAb-IgG, the cAMP production and 3H-thymidine incorporation in FRTL-5 cells were much higher than those from normal controls (P < 0.05 respectively). Using 32P labelled v-fos probe by the Northern Blot method, the expression of c-fos mRNA could be induced by IgGs from patients with GD. CONCLUSIONS: These data suggest that the stimulation of TRAb-IgG followed by cAMP production and 3H-thymidine incorporation is related to the induction of c-fos mRNA and, thus, to the growth of FRTL-5 cells. PMID- 10374351 TI - Video-assisted cardiac surgery: preliminary results in Chang Gung Memorial Hospital. AB - OBJECTIVE: To summarize the experience of utilization of video-assisted endoscopy in 91 patients operated on at Chang Gung Memorial Hospital, Taipei, China. METHODS: From October 1995, through August 1996, 91 patients (44 male and 47 female) received video-assisted cardiac surgery (VACS). Their ages ranged from 1 year to 79.5 years (25.7 +/- 21.7). Indications for surgery were atrial septal defect (59 patients), ventricular septal defect (15), coronary artery disease (4), severe mitral regurgitation (4), severe tricuspid regurgitation (3), thrombosis of mitral mechanical prosthesis (3), left atrial tumor (2), and left ventricular thrombus with dilated cardiomyopathy (1). The VACS was performed through right or left anterior minithoracotomy and guided by video-assisted endoscopic techniques by means of projected images on the video monitor under extracorporeal circulation. The aorta was not cross-clamped and the myocardium was protected by continuous coronary perfusion with hypothermic fibrillatory arrest (rectal temperature 22.6 +/- 4.0 degrees C). Conventional instruments were used. RESULTS: All lesions were corrected successfully. The bypass time was 27 to 335 minutes (72.8 +/- 52.7). The operative time was 1.3 to 8.5 hours (3.0 +/- 1.7). There were no operative deaths and 3 late deaths. Follow-up was complete in all survivors (6 to 16 months, mean 8.7). Most of them were found to be in NYHA functional I or II. CONCLUSION: Our preliminary experiences demonstrate that VACS is simple and effective in surgical correction of selected cardiac lesions. Short term results show good outcomes. PMID- 10374350 TI - Genetic heterogeneity for familial hypertrophic cardiomyopathy in Chinese: analysis of six Chinese kindreds. AB - OBJECTIVE: Familial hypertrophic cardiomyopathy (FHCM) is a primary myocardial disease characterized by unexplained ventricular hypertrophy. The application of the techniques of reverse genetics has identified at least five chromosomal loci as the major causes for FHCM in diverse ethnic populations, suggesting substantial genetic heterogeneity for FHCM. Recently, the defective gene loci of two Chinese families with FHCM have been mapped to chromosome 11 and 14q1, respectively. For further understanding of the molecular basis of FHCM in Chinese, we analyzed the linkage between four other Chinese kindreds and DNA markers from chromosome 14q1. METHODS: Six unrelated Chinese families with FHCM, including two previously reported, were studied. Totally 90 family members were included for analysis. DNA from 80 individuals was extracted and polymerase chain reactions were performed using the primers designed according to the sequences derived from the alpha and beta myosin heavy chain gene. Totally four polymorphisms were studied, including three polymorphic microsatellite sequences and one single strand conformation polymorphism. Genetic linkage analysis were performed using the Linkage program. RESULTS: In the six studied families, 39 of the 90 family members were found to be affected diagnosed either by echocardiography or by clinical evaluation. The pattern of inheritance in all six studied families was most consistent with an autosomal dominant trait with a high degree of penetrance. Genetic linkage analysis using polymorphisms on the alpha and beta MHC genes showed a combined maximal lod score of 6.2 for trinucleotide repeat polymorphism AMHC-I 15 at theta = 0.00 for three studied families without recombination. Exclusion of linkage to the chromosome 14q1 location was noted in two of three other families with the maximal lod score of -2 or less. CONCLUSIONS: These results provide further evidence that FHCM in Chinese is genetically heterogeneous. Chromosome 14q1 locus, probably the beta myosin heavy chain gene, is important as the molecular basis for FHCM in Chinese. PMID- 10374352 TI - Electron microscopic observations of apoptotic cells in various etiologies of human cardiovascular diseases. AB - OBJECTIVE: To observe apoptotic process in various cardiovascular disorders with a particular attention to the ultrastructural morphology of apoptosis in cardiomyocytes, fibroblasts, vascular endothelial cells and smooth muscle cells. METHODS: Transmission electron microscopic observations of the tissue specimens obtained from endomyocardial biopsies or surgical excisions of left ventricular myocardium or calcified aortic valves were carried out in 50 patients with various cardiovascular diseases. RESULTS: The ultrastructural features of apoptosis was consistently observed in cardiomyocytes, fibroblasts, vascular endothelial cells and smooth muscle cells in all diseased tissues. In cardiomyopathies and rheumatic heart diseases apoptosis was commonly observed in the cardiomyocytes. It was often found that fibroblasts underwent apoptosis in calcific aortic valve tissues. Apoptosis of arterial smooth muscle cells was a frequent finding in renal arterial stenosis due to Takayasu's arteritis and fibromuscular dysplasia. Regardless of the cell types, the nuclear hallmarks of apoptosis were identical with minor modifications of the fragmentation of the condensed cells into apoptotic bodies. CONCLUSIONS: Based on electron microscopic findings, it is suggested that the underlying disease processes determine which type of cells predominantly undergoes apoptotic changes in various cardiovascular disorders. In addition, different cells with similar structural morphology may have common ultrastructural features of apoptosis. PMID- 10374353 TI - Psychological interventions in general hospitals: background, current status and clinical guidelines. AB - PURPOSE: To promote the systematic development, interests, practice, research and clinical applications of health psychology in general hospitals in Hong Kong and the mainland of China. DATA SOURCES: The targets and aims of therapeutic work with patients in pain, cancer patients, child and adolescent patients, patients with chronic illnesses, the elderly, and patients requiring organ transplantation are highlighted. STUDY SELECTION: The psychological interventions described are experiences derived from routine clinical services carried out in the Clinical Health Psychology Unit where the authors are affiliated, and can be seen as an example of a more comprehensive psychological intervention program for physically ill patients in Hong Kong. RESULTS: Psychological interventions have intrinsic values in reducing patients' distress and sufferings. The services are also an integral part of modem day comprehensive patient care with positive effects on treatment effectiveness and eventual illness outcome. CONCLUSIONS: Physical illnesses affect a person physically as well as psychologically. Psychological care in general hospitals is cost effective and beneficial in reducing undue psychological complications precipitated by physical afflictions as well as in promoting better overall outcomes. PMID- 10374354 TI - Histological and electron-microscopic observations on the mucosa of pediculated duodenal wall graft transplanted to the stomach in Wistar rats. AB - OBJECTIVE: To study the mechanism of gastric metaplasia in duodenum through the transplantation of a flap of duodenal wall with vascular pedicle to the stomach. METHODS: The pediculated duodenal wall flaps of Wistar rats were transplanted to their stomachs. The rats were killed at the 3rd, 6th, 9th and 12th month after the operation respectively, and histological changes of the duodenal grafts were observed with optical and electron microscopy. RESULTS: Gastric metaplasia was found in the mucosa of duodenal grafts transplanted to the stomach at the 6th, 9th and 12th month. CONCLUSIONS: The formation of gastric metaplasia in the duodenal mucosa may be related to a change of the microenvironment around the tissues, and duodenal mucosa may differentiate into gastric epithelium by the decrease of pH value. PMID- 10374355 TI - Synergetic protective effects of combined blockade by two kinds of autolesion mediator receptor on neurological function after cervical cord injury. AB - OBJECTIVE: To investigate the effects of combined blockade by platelet activating factor (PAF) receptor antagonist BN52021 in combination with opiate receptor antagonist naloxone on neurological function and neurological tissue damage after cervical cord injury. METHODS: Spinal cord contusion at C6 segment was made with Allen method in cats, which were randomly divided into four groups: saline control group; BN52021 group; naloxone group; and combined treatment group with BN52021 and naloxone. Alteration of cervical cord blood flow, blood barrier permeability of the spinal cord, cervical cord tissue pathology and neurological functional scores were studied after experimental cervical cord injury. RESULTS: The animals treated with BN52021 or naloxone had significantly better functional scores than saline controls 6 weeks after injury (P < 0.05). Moreover, the combined treatment showed significantly better neurologic recovery than either naloxone or BN52021 treated animals (P < 0.05). The other indexes in combined treatment animals were also superior to those in naloxone or BN52021 treated animals. CONCLUSIONS: Combined blockade by two kinds of autolesion mediator receptor can more effectively inhibit secondary damage production and development after cervical cord injury and improve neurologic function. PMID- 10374356 TI - Dynamic stability of glenohumeral joint during scapular plane elevation. AB - OBJECTIVE: To investigate the muscle-controlled dynamic stability of the glenohumeral joint through X-ray fluoroscopy in active and passive shoulder elevation in scapular plane. METHODS: Sixty healthy volunteers were collected in this study, including 23 men and 37 women, with an average age of 28.4 years. Passive and active shoulder elevation in scapular plane were observed under X-ray imaging. In 18 subjects, X-ray films were taken when the shoulder elevated in scapular plane from 0 degree to 150 degrees with a 30 degrees interval at each stage in both active and passive movements. The angles between the pivot of the humerus and the glenoid surface (GHA) during the active and passive motion were calculated and analyzed. Manual examination was also applied in the same manner. RESULTS: The pivot of the humerus had a tendency to be vertically closer to the glenoid surface in the active elevation than in the passive elevation. The differences of GHA between the active and passive motion at 0 degree, 30 degrees, 60 degrees, 90 degrees, 120 degrees and 150 degrees elevation were 4.55 degrees +/- 0.37 degree, 5.44 degrees +/- 1.16 degrees, 6.50 degrees +/- 1.50 degrees, 4.94 degrees +/- 0.82 degree, 4.50 degrees +/- 0.40 degree and 1.44 degrees +/- 0.68 degree, respectively. Manual examination found the angle between the scapula and the humerus tended to be larger in the active motion than in the passive motion. CONCLUSION: The active coordination of the muscles around the shoulder is beneficial to the dynamic stability of the glenohumeral joint. PMID- 10374357 TI - Experimental model of renovascular hypertension. AB - OBJECTIVE: To establish a model of renovascular hypertension. METHODS: A 4/0 resorbable chromic catgut ligature was used to ligate subtotally the renal arteries of 18 dogs, forming experimental renovascular hypertension steadily. Blood pressure, plasma renin activity, the ultrastructural changes of juxtaglomerular apparatus and renal artery wall were studied after the constriction. RESULTS: It was reasonable that renal blood flow measured with an electromagnetic flowmeter was reduced by 30% after the constriction. The pathological changes of the induced renal artery stenosis were similar to those of fibromuscular dysphasia. CONCLUSION: The findings provide valuable evidence for the treatment of renovascular hypertension. PMID- 10374358 TI - Intrahepatic peripheral cholangiocarcinoma: CT features in 18 pathologically proven cases. AB - OBJECTIVE: To determine the morphological features of 18 pathologically proven intrahepatic peripheral cholangiocarcinoma (IHPCC) cases in computerized tomography (CT) image. METHODS: All 18 patients had CT, using Picker I.Q.T/C and taking pre-contrast continuous 10-mm sections throughout the liver and post contrast continuous 10-mm sections throughout the focuses. RESULTS: The disease was characterized in CT image by the following: all focuses were found in the periphery of the liver and shown as a lobulated or fused hypodense space occupying mass; there were one or more divergent or confluent, circular or irregular cystic areas with much lower density, in the majority. All focuses could be enhanced slightly and most revealed a dim edge. Dilated bile ducts around the focus were found frequently; the dilated bile ducts especially seemed to encircle the focus (33.3%, 6/18). This phenomenon were referred to as "encysted sign of dilated bile ducts". CONCLUSIONS: CT scanning should be one of the most important investigative methods for IHPCC due to the disease characteristics identified in CT image, especially "encysted sign of dilated bile ducts" which possesses specificity in diagnosing the disease. PMID- 10374359 TI - Abdominal aortic aneurysm in Hong Kong: audit from a teaching hospital (1975 1995). AB - OBJECTIVE: To analyse the epidemiology and trends of abdominal aortic aneurysm (AAA) in an ethnic Chinese population over a 21-year period in a representative referral institution in Hong Kong. METHODS: Over a 21-year period from 1975 to 1995, 533 patients with abdominal aortic aneurysms were treated in a single Vascular Surgical Unit at the Department of Surgery, Queen Mary Hospital, Hong Kong. Their characteristics, treatment and mortality were reviewed. RESULTS: There was an increase in the incidence from a mean annual admission of 10 cases per year during 1975-1980, to that of 38 per year during 1991-1995. The increase was largely detected in males, the male to female ratio being 2.5:1 in the 1980s and 3.6:1 in the 1990s. There was no parallel increase in the incidence of ruptured aneurysms which accounted for 12.6% of all aneurysm admissions. CONCLUSION: There is a progressive increase in the incidence of AAA in Chinese patients, in a time frame and pattern similar to that reported in the West. PMID- 10374360 TI - The basic clinical diagnostic framework synergized. AB - OBJECTIVE: To develop a diagnostic framework that would help defining clinical problems by the expanded understanding with traditional Chinese medicine (C Med) and western medicine (W Med). METHODS: The basic attributes of diagnosis and the use of diagnostic techniques of W Med and C Med are reviewed. Some of the various diagnostic labels and their meanings are also preliminarily reviewed. A consolidatory approach is made to synergize the usefulness of the two disciplines based on the principles of the two schools. RESULTS: A synergized basic diagnostic framework is developed. The disease (the disease diagnosis, [symbol: see text]), the state of the inner core (the core diagnosis, [symbol: see text]) should be defined. The term "systemic clues" is used to describe the symptom complex and conglomerate evidence describing the transient or sustained reaction of the inner core to environment. This is an analytical summary of its pathophysiological changes in reaction to the environmental insults. CONCLUSIONS: The application and use of the framework are discussed and propositions are made. The synergized diagnostic platform represents a starting effort to form a unified basis to exchange clinical diagnosis between W Med and C Med. Such framework may facilitate looking for new treatment modalities and results can be compared across different centres. PMID- 10374361 TI - Fifteen years' review of advanced childhood neuroblastoma from a single institution in Hong Kong. AB - OBJECTIVE: To assess the progress in the treatment of advanced childhood neuroblastoma. METHODS: From 1981 to 1996, there were 32 children with neuroblastoma (NB) diagnosed, staged and treated in our institution. There were 4 patients with stage II NB (12%), 5 stage III (16%), 21 stage IV (66%) and 2 stage IV s (6%). The NBs were excised if CT scan indicated that the tumors were operable. For advanced NB, stages III and IV, multiple drug chemotherapy was started first and operability was assessed with serial CT scan examinations. Once the X-ray imaging indicated the tumors were operable, surgical interventions were done. The medical records of the advanced NB were reviewed. RESULTS: In the initial period of the study, 9 patients were treated using the VAC protocol [vincristine (vcr), adriamycin (adria) and cyclophosphamide (cyc)]. No patient was convertible to operable and all died with a mean survival of 10 months. OPEC [vcr, cyc, VM26, cisplatin (cis)], Rapid COJEC (carboplatin, VP16, vcr, cis and cyc) and more recently N6 protocol (cyc, adria, vcr, VP16, cis) was used for 17 patients. 80% of them were converted to operable. In 4 patients, surgical specimens showed only necrotic tissue without viable tumor tissue and 6 (35%) tumors were converted to ganglioneuroma or ganglioneuroblastoma. Although 2 (12%) patients died of fungal septicemia and 1 (6%) developed Fanconi's syndrome after chemotherapy, the mean survival period increased to 27 months. In the 10 survivors (60%), 4 had megatherapy with melphalan followed by autologous peripheral blood stem cell (PBSC) transplantation and 2 were waiting for transplantation. CONCLUSIONS: There is a high percentage of advanced NB on presentation in Hong Kong. With more potent multiple drug chemotherapy for advanced stage NB there are (1) improvement in the survival of these patients, (2) opportunities for more operations for tumor excision and (3) opportunities for autologous PBSC transplantation for better tumor eradication. PMID- 10374362 TI - Alveolar echinococcosis in China. AB - DATA SOURCES: All reference data originated from related Chinese- or English language literature in Chinese journals. STUDY SELECTION: Twenty-three original articles published in 1992-1996 were selected according to the stated purpose and 9 of them were written by myself. DATA EXTRACTION: The present paper dealt with 5 subtopics, i.e. epidemiology, parasitology, pathology, diagnosis, treatment and prognosis. RESULTS: Five hundred and eighty-four patients with alveolar echinococcosis (AE) have been reported from 7 provinces or autonomous regions. Human infection rate was 19.2% or 2.8%, and the morbidity was 2.4%-5.0%. The intermediate hosts of Echinococcus multilocularis included 7 species of rodents and 3 species of livestocks, and the final hosts of that were fox, dog or wolf. Diagnosis of AE was chiefly based on imaging examination (ultrasound and CT) and immunological test. The operative resection rate for liver AE was only 10.5% (27/258). Albendazole was a certain remedy for the chemotherapy of AE, and TCM "Xiao-Bao" pill may be a hopeful drug. CONCLUSIONS: In the past 30 years, a lot of work concerning basic and clinical studies has been done in China and some achievements have been achieved. However, some important problems such as how to conduct further research on molecular biology, how to make early diagnosis, how to improve the chemotherapeutic effect, and how to control the prevalence of AE, need to be studied more deeply in the 21st century. PMID- 10374363 TI - Atrial pacing lead: steroid-eluting and nonsteroid-eluting. PMID- 10374364 TI - Relationship between angiotensin I converting enzyme gene polymorphism and diabetic nephropathy. PMID- 10374365 TI - Strengthening research on trauma. PMID- 10374366 TI - Study on delay two-phase multiple organ dysfunction syndrome. AB - OBJECTIVE: To study the injury factors, pathogenic process and clinical features of delay two-phase multiple organ dysfunction syndrome (MODS) in severe burned patients and to replicate a standardized animal model that would accurately imitate the clinical features of MODS. METHODS: Forty-five human patients with burn size larger than 30% total body surface area (TBSA) were analyzed. All of them underwent severe burn shock in early stage and sepsis in late stage. Thirty two goats were randomly divided into three groups: 1) hemorrhagic shock (group H, n = 6); 2) endotoxemia (group E, n = 6); and 3) hemorrhagic shock plus endotoxemia (group M, n = 20). Hemorrhagic shock was produced according to the method of Wigger (6.7 kPa for an hour, 1 kPa = 7.5 mmHg). Endotoxin (E. coli O111 B4) was given via the portal vein 24 hours after the resuscitation of hemorrhagic shock, in a dose of 30 ng/kg/min for 5 consecutive days. During the observation period of 10 days, all animals were hemodynamically monitored, given standard metabolic support and due cardiac and pulmonary support according to human intensive care. RESULTS: All the patients showed burn shock at 1-3 days and hyperdynamic circulation, hypermetabolism and systemic inflammatory responses over two weeks post-injury. Thirteen cases were found to develop MODS according to the prevailing diagnostic criteria, and 10 of them died with a mortality of 77%. Eighteen animals died in group M with a mortality of 90%, 12 of the 18 developed MODS, with overall incidence of 60%. Most animals in group M showed changes similar to that observed in human cases. The experimentation proved that in the pathogenic process of MODS, there was a two-hit phenomenon in the dvelopment of the syndrome. To prevent the development of MODS, it therefore was imperative to blunt the first hit or the second hit, so that an excessive inflammatory response was alleviated. This postulation has been verified in the treatment of extensive burns. Two patients with burn extent reaching 100% TBSA survived with only mild acute respiratory distress syndrome (ARDS) and renal dysfunction after comprehensive treatment of burn shock, including adequate fluid resuscitation, drugs to remove oxygen free radicals, rapid restoration of pHi, and early extensive excision of burn eschars. CONCLUSION: Both in human patients or animal experimentation, the typical delay two-phase MODS is shown to be produced by two successive insults in the forms of hypovolemic shock and sepsis. This postulation is helpful in formulating the prevention and treatment modality of MODS. PMID- 10374367 TI - Development of serial bio-shock tubes and their application. AB - OBJECTIVE: To design and produce serial shock tubes and further examine their application to experimental studies on blast injury. METHODS: Bio-medical engineering technique was used for the design and development of the serial shock tubes. One thousand four hundred and fifty nine animals (757 rats, 105 guinea pigs, 335 rabbits, 240 dogs and 22 sheep) were then used to test the wounding effects of the shock tubes. RESULTS: Three types of bio-shock tubes, that is, large-, medium- and small-scale shock tubes were made in our laboratory. The large-scale shock tube is 39 meters long; the inner diameter of the test section is 1 meter; and the maximum overpressure in the driving section is 10.3 MPa. A negative pressure could be formed by means of the reflected rarefactive wave produced by the end plate. The medium-scale shock tube is 34.5 meters long; the maximum overpressure in the driving section is 22 MPa; the test section is designed to be a knockdown, showing 5 basic types with inner diameter of 77 to 600 millimeters, which could be used for researches on overpressure, explosive decompression, underwater explosion, and so on. The small-scale shock tube is 0.5 meter long with the maximum endured overpressure of 68.6 MPa. Results from animal experiments showed that this set of shock tubes could induce various degrees of systemic or local blast injury in large or small animals. CONCLUSIONS: This set of bio-shock tubes can approximately simulate typical explosive wave produced by nuclear or charge explosion, and inflict various degrees of blast injury characterized by stability and reproducibility. Therefore, they can meet the needs of blast research on large and small animals. PMID- 10374368 TI - Experimental study on firearm wound in maxillofacial region. AB - OBJECTIVE: To make clear the range of firearm wound in the maxillofacial region, the optical repair time and the characteristics of accompanied indirect brain damage, and to offer the principle of emergency treatment and the early repair of war wound. METHODS: With the aid of the standard Sweden model, 200 dogs were used in the experiment. Varies tissues around the primary canal were harvested chronologically, in different zone and different tissue, for histopathological examination. RESULTS: The necrotic range of various tissues in the maxillofacial region was less than that in the extremities. In the maxillofacial region, there was a significant temporary cavity following the passing of bullet, which caused indirect brain damages. CONCLUSION: These findings are helpful to the treatment of war wound in the maxillofacial region. Early bone transplantation using microvascular anastomosis in the treatment of gunshot wound in the maxillofacial region is recommendable. PMID- 10374369 TI - Distribution of endotoxins in tissues and circulation and its effects following hemorrhagic shock. AB - OBJECTIVE: To systemically investigate 1) distribution of endogenous endotoxin (ET) in tissues and circulation; 2) its relationship with shock duration and organ damage; and 3) its possible mechanism after hemorrhagic shock. METHODS: To further elucidate the intrinsic relationship between endogenous endotoxin translocation and hemorrhagic shock, the present study systematically investigated the distribution of endogenous ET into the liver, lungs, kidneys and circulation, and the relationship between ET levels and the corresponding organ dysfunction with limulus amebocyte lysate (LAL) chromogenic assay following hemorrhagic shock in rats. RESULTS: It was found that ET levels in hepatic homogenate markedly increased (P = 0.09) 1.5 hours following shock compared with that in the sham group. After resuscitation, ET levels in hepatic, pulmonary and renal tissues were all significantly elevated. The levels kept increasing with the prolonged experimental time, and reached as high as 3.88 +/- 0.95 EU (endotoxin unit)/g in the livers, 2.53 +/- 1.46 EU/g in the lungs and 2.51 +/- 0.89 EU/g in the kidneys 12 hours after shock. ET levels in plasma reached a peak of 1.13 +/- 0.42 EU/ml at 1 hour following resuscitation, then rapidly decreased to the sham levels 3 hours following resuscitation. There was a close relationship between endotoxin translocation and shock duration. Correlation analysis further indicated that the changes in glutamic-pyruvic transaminase (GPT), blood urea nitrogen (BUN) in plasma and angiotensin I-converting exzyme (ACE) in pulmonary homogenate were significantly and positively correlated with the ET levels in the liver, kidneys and lungs after hemorrhagic shock. CONCLUSIONS: Hemorrhagic shock can induce obvious endogenous ET translocation, which is closely related to the shock duration. Although only transient endotoxemia occurs after hemorrhagic shock, ET can massively accumulate in tissues (liver, lungs and kidneys), and may play an important role in the development of shock. PMID- 10374370 TI - Preventive effect of artemether on schistosome infection. AB - OBJECTIVE: To study the preventive effect of artemether (Art) in protecting the people from schistosome infection during flood fighting in schistosomiasis endemic area of Poyang Lake, Jiangxi Province. METHODS: From mid July to mid August in 1996, the water level in Poyang Lake rose due to torrential rains and 2 embankments, Zhedi and Jiangtongdi, which appeared in dangerous situation and were selected as the pilot spots. After those who went to fight against flood arrived at the pilots their sera were collected within 48 hours and were examined with indirect hemagglutination test (IHA), enzyme-linked immunosorbent assay (ELISA) and McAb-ELISA. Individuals with negative outcome in the 3 tests were then selected as the study subjects and were allocated randomly to the Art or the control group. The first dose of Art given to the individuals contacted with the infested water within 11-15 days was 6 mg/kg. If the individual continually contacted the infested water, the same dose of Art was given once every 15 days. After the individuals withdrew from the pilot, one more dose of Art was administered 7-15 days later. Placebo (starch) was given to individuals in the control group at the same period as in artemether group. Stool examinations were made in both groups 40-50 days after the last medication for evaluation of the preventive effect of artemether. Double blind method was used in the administration of both artemether and placebo. RESULTS: In Zhedi pilot, the individuals fought against flood for about 1 month. In Art group, 99 individuals receiving 3 doses of the drug completed the stool examination with egg-positive rate of 4% and no acute schistosomiasis was seen. In the control group, among 110 people who completed the observation, 44 were egg-positive with an infection rate of 40%, and 29 were identified as having acute schistosomiasis. In Jiangtondi, the studied individuals contacted the infested water for only about 4 hours. But in the control group 4 out of 102 individuals were egg-positive, while none of the 103 individuals in Art group receiving 2 doses of the drug showed schistosome infection. No apparent side effect was seen in the people treated with artemether. CONCLUSION: After oral Art was given to the people fighting against flood in schistosomiasis endemic area of Poyang Lake, it was shown that the oral Art has a promising effect on controlling acute schistosomiasis and reducing the infection rate. PMID- 10374371 TI - Study of hepatitis C virus genotype in Guizhou area of southwestern China. AB - OBJECTIVE: To determine the concordance in various hepatitis C (HCV) genotyping methods and to investigate the distribution of HCV genotypes in Guizhou area of Southwest China. METHODS: Serum samples from 206 patients (100 with chronic hepatitis and 106 with hemopathy) were detected for antibody of HCV by second generation enzyme-labelled immunosorbent assay (ELISA). Thirty-five anti-HCV positive samples were detected for HCV RNA by RT-polymerase chain reaction (PCR) and 30 HCV RNA positive samples were determined for their genotypes by three various genotyping methods [PCR with type-specific primers at the core region (primer-set), slot-blot hybridization with type-specific probes at NS5B region (blotting) and the restric fragment length polymorphism analysis of PCR products of 5' NC region (RFLP)]. Ten samples with the known genotype were analysed by the direct sequencing. RESULTS: Of 30 samples with positive HCV RNA, the types of 22 could be classified by three methods, and the genotypes determined by various methods had complete concordance. The types of 6 samples could be classified by two methods and 5 had agreement subtypes. The types of two samples could be classified only by RFLP. Overall, 27 (90.0%) had subtype 1b infection and 3 (10.0%) had subtype 2a infection. The nucleotide sequence of 8 samples with subtype 1b and one with subtype 2a were analysed by the direct sequencing. The subtypes determined by sequence analysis were in complete concordance with those decided by various genotyping methods. CONCLUSIONS: Subtype 1b is the predominent HCV genotype in Guizhou area, while subtype 2a is less common. There was a good concordance with the genotyping results obtained by various HCV genotyping methods. PMID- 10374372 TI - Effects of different subtypes of histamine receptors on proliferation and differentiation of murine colony forming unit granulocyte-macrophage and colony forming unit megakaryocyte. AB - OBJECTIVE: To investigate the function and characteristics of histamine receptors on the hemopoietic progenitor cells. METHODS: BDF1 mice (both male and female), inbred at our university, aged 8-12 weeks, weighing 20-24 g, were used in this study. Bone marrow cells were incubated for 1 hour at 37 degrees C with 2-AT (H1 receptor agonist) or impromidine (H2 receptor agonist) alone, or in combination with the antagonists pyrilamine or cimetidine respectively. Control experiment was performed in Dulbecco's modified Eagle's Medium (DMEM) alone. Cells treated with different drugs were performed by colony forming unit-granulocyte-macrophage (CFU-GM) and colony forming unit-megakaryocyte (CFU-Meg) assay. RESULTS: When bone marrow cells were treated with 10(-8) mol/L to 10(-5) mol/L of 2 thiazolylethylamine (2-AT) which had no influence on CFU-GM and CFU-Meg proliferation, 10(-8) mol/L to 10(-5) mol/L of impromidine could increase the number of CFU-GM and CFU-Meg colonies. The effects of H2 receptor agonists on CFU GM, CFU-Meg could be antagonized by H1 receptor agonist. CONCLUSIONS: Our findings suggest the existence of histamine H1 and H2 receptors on the hemopoietic progenitor cells and the antagonism between two different histamine receptor subtypes on the proliferation of CFU-GM and CFU-Meg. PMID- 10374373 TI - Changes of skin perfusion after photodynamic therapy for port wine stain. AB - OBJECTIVE: To obtain an objective assessment of the curative effectiveness of photodynamic therapy (PDT) for port wine stain (PWS), we investigate the relationship between the microvascular perfusion changes of PWS and the blanching of the lesions before and after PDT. METHODS: Twenty-four patients (18 females and 6 males with a total of 28 lesions) suffering from PWS were treated with PDT. The lesions of various extents were located on the face and neck. After intravenous injection of photosensitizer hepatoporphyrin derivative (HpD), the copper vapor laser was adopted as light source and the lesions of PWS were irradiated. The laser Doppler perfusion imager (LDI) was used to measure the microcirculatory perfusion of PWS before and after PDT and comparison with the normal skin was done. RESULTS: All the lesions showed remarkable decrease of tissue perfusion after PDT. It was shown that the mean, maximal and minimal values of tissue perfusion in the pre-treatment group were significantly higher than those in control group (P < 0.01). Six months after PDT, the mean, maximal and minimal values of perfusion with the lesions were reduced, with significant difference from pre-treatment group (P < 0.01), but no significant difference from control's. The colors of lesions were correlated with decrease of microcirculatory perfusion, which became lightened close to normal skin color without causing any scarring. CONCLUSIONS: PDT is one of the most effective modalities for PWS. The microcirculation perfusion can reflect the degrees of PWS objectively. The curative effectiveness of PDT for PWS is due to tissue microcirculation response. PMID- 10374374 TI - Cardiac protection by long-term treatment with captopril in patients with acute myocardial infarction. AB - OBJECTIVES: To assess the effects of long-term angiotensin-converting enzyme (ACE) inhibitor treatment with captopril on cardiac function in acute myocardial infarction (AMI). METHODS: One hundred and one patients with AMI who were admitted to hospital within 72 hours of the onset of symptoms with no cardiogenic shock were randomly allocated to captopril (n = 52; group I) and conventional treatment (n = 49; group II). Left ventricular (LV) systolic performance and diastolic transmitral flow velocity profiles were assessed by Doppler echocardiography at admission (1.2 +/- 1.1 days), before discharge (27 +/- 10 days) and during follow-up (363 +/- 31 days). RESULTS: At one year follow-up, in group I LV end-diastolic volume decreased, and ejection fraction increased due to a disproportionate decrease in end-systolic volume. The incidence of cardiac dilatation was reduced. LV early diastolic filling velocity (E) increased and late atrial filling velocity (A) decreased, resulting in an elevation of E/A ratio. However, the mean values of LV systolic and diastolic functional parameters were unchanged in group II. CONCLUSIONS: Long-term treatment with captopril exerts a beneficial effect on cardiac protection for patients with AMI. PMID- 10374375 TI - Beneficial effects of early coronary reperfusion on left ventricular remodeling and systolic function in patients with acute myocardial infarction. AB - OBJECTIVE: To evaluate the beneficial effects of early coronary reperfusion on left ventricular remodeling (LVRM) and systolic function in patients with acute myocardial infarction (AMI). METHODS: Eighty-one patients with first AMI in the convalescent stage and having undergone left ventriculography (LVG) and coronary arteriography (CAG) were divided into four groups: the anterolateral wall (ALW) myocardial infarction (MI) non-reperfusion (n = 20) and reperfusion (n = 21), and inferoposterial wall (IPW) MI non-reperfusion (n = 20) and reperfusion (n = 20), according to infarct location and early treatment with or without successful coronary reperfusion therapy within 6 hours after onset of symptoms. By LVG, the parameters of LVRM and systolic function in the four MI groups were analyzed and compared with those in normal group (n = 25) and between the two reperfusion and non-reperfusion MI groups. RESULTS: In both ALW and IPW MI non-reperfusion groups, the left ventricular (LV) end-diastolic volume (EDV), circumference (EDC), short-axis dimension (EDD), short to long axis ratio (ED-D/L), sphericity index (ED-SI) and end-systolic volume (ESV) were all significantly increased (P < 0.01-0.001), while LV ejection fractions (LVEF) were significantly decreased (both P < 0.001) when compared with those of normal group; and the increase in ESV and decrease in LVEF were both significantly greater in ALW than in IPW MI groups (both P < 0.01). In both ALW and IPW MI reperfusion groups, however, the EDV, EDD, ESV, as well as the extent and severity of regional wall motion abnormality (RW-MA) were significantly smaller (P < 0.05-0.001), while LVEF were significantly higher (P < 0.01-0.001) when compared with those in the two non reperfusion MI groups respectively. There were no longer significant differences in LVEF and ESV between ALW and IPW MI groups (both P > 0.05). The EDC in IPW MI reperfusion group and the ED-D/L and ED-SI in ALW MI reperfusion group were also significantly reduced compared with those in the two non-reperfusion MI groups respectively (P < 0.05-0.001). All the above parameters in the two reperfusion MI group were decreased to the normal in comparison with normal group except ESV and LVEF, and ED-D/L and ED-SI in IPW MI group. CONCLUSION: It was indicated that in both ALW and IPW MI non-reperfusion groups, LVRM had occurred in convalescent stage of AMI with an increase in EDV and EDC, spherical change in LV shape, and accompanying reduction in LV systolic function; and early coronary reperfusion in AMI could reduce the extent and severity of RWMA, prevent from LV enlargement and remodeling, and preserve or improve LV systolic function with more prominence in ALW MI. PMID- 10374376 TI - Erythrocyte and plasma Ca2+, Mg2+ and cell membrane adenosine triphosphatase activity in patients with essential hypertension. AB - OBJECTIVE: To assess the relationships between plasma and intracellular Ca2+, Mg2+ and blood cell membrane adenosine triphosphatase (ATPase) activity in normotensive and hypertensive subjects. METHODS: Plasma and intracellular Ca2+, Mg2+ were measured with atomic absorption spectrophotometry, and red blood cell membrane Na(+)-K+ ATPase and Ca(2+)-ATPase activities were determined with colorimetric method in 55 patients with essential hypertension and 32 normotensive controls. RESULTS: The results showed that the hypertensive group consistently demonstrated a significant decreased activity of ATPase studied, with significantly lower plasma Ca2+ and higher cytosolic Ca2+ levels when compared with those in normotensive group (P < 0.01 or P < 0.05, respectively). No significant differences were found in either plasma Mg2+ or intracellular Mg2+ level between the two groups. CONCLUSIONS: This study suggests that patients with essential hypertension have widespread depression of cell membrane Na(+)-K(+) ATPase and Ca(2+)-ATPase activities with plasma Ca2+ depletion and cytosolic Ca2+ overload, which may reflect an underlying membrane abnormality in essential hypertension. The cellular abnormalities may be related to the defective transport mechanisms that in turn may be aggravated by plasma Ca2+ depletion. PMID- 10374377 TI - Radioimmunoassay of human cardiac acidic isoferritin: a new index for hepatic cancer. AB - OBJECTIVE: To investigate human cardiac acidic isoferritin as a specific index of hepatic cancer. METHODS: Acidic isoferritin was isolated and purified from human heart muscle. A radioimmunoassay for the acidic isoferritin in human serum has been developed, on the equilibrium method. The antiserum was obtained from rabbits immunized with purified acidic isoferritin. The 125I-acidic isoferritin was prepared by the chloramine-T method. The data were processed using the automated smoothed spline function data processing program. RESULTS: The intra- and inter-assay CV of acidic isoferritin RIA were 1.65% and 9.71%, respectively, and the recovery rate was 102%. The antiserum provided a linear response from 7.0 to 369.6 micrograms/L with ED50 of 27.50 micrograms/L. The cross reactivity with AFP, CEA, lactoferrin and transferrin was negligible, and that with ferritin was 1.74%. The serum acidic isoferritin concentration showed a considerable variation in different sex and age groups. The serum acidic isoferritin was measured in liver diseases including hepatic cancer, hepatic cirrhosis and acute and chronic hepatitis. Its sensitivity for diagnosis of hepatic cancer was 73.05%, independent from the severity of hepatic injury. In 8 malignant tumors studied, acidic isoferritin appeared the most valuable in the diagnosis of hepatic cancer, with its positive, negative, false positive and false negative rates all being ideal. CONCLUSIONS: Acidic isoferritin may turn to be a rather specific index of hepatic cancer. Combination of monitoring both acidic isoferritin and AFP would raise the positive detection and specificity in the diagnosis of hepatic cancer. PMID- 10374378 TI - An experimental study on rat model of parkinsonism by gene therapy. AB - OBJECTIVE: To induce significant improvement of motor abnormalities and striatal dopamine (DA) levels in rat model of Parkinson's disease (PD), by intracerebral grafting of the genetically modified muscle cells expressing tyrosine hydroxylase (TH). METHODS: Primary myoblasts and myotubes from the rat were prepared by cell culture and a plasmid, pCMVTH, containing TH gene and a promoter of cytomegalovirus (CMV) was constructed by DNA recombination technique. The primary muscle cells were transfected with newly constructed pCMVTH DNA vector, by using lipofection. These genetically modified muscle cells were grafted into the caudate- putamen of 6-OHDA-lesioned rats, representing PD models. Before and after grafting, the rotational behaviour and the striatal levels of DA and its metabolities were tested at different postoperative survival times. In addition, the immunocytochemistry for showing TH activity was done both in vitro and in vivo. RESULTS: The newly constrcuted plasmid, pCMVTH was proved to contain TH gene and have correct direction of insertion. The cultured primary myoblasts and myotubes lipofected with pCMVTH were immunocytochemically shown to express TH activity in vitro. After grafting, these TH-expressing muscle cells showed to have a long-term survival cells in vivo and induced a marked decrease in abnormal locomotion and a increase in striatal DA levels for PD rat model. CONCLUSIONS: In experimental gene therapy for PD, the pCMVTH is a useful vector for carrying TH gene. The lipofection is a practical technique for transferring a target gene into eukaryotes and primary cultured muscle cells should be a good vehicle for DNA transfer and intracerebral grafting. PMID- 10374379 TI - Trinucleotide repeat expansion of spinocerebellar ataxia (SCA1) found in a Chinese family. AB - OBJECTIVE: To investigate the gene mutation and the ratio of the spinocerebellar ataxia type 1 (SCA1) in Chinese patients with autosomal dominant spinocerebellar ataxia (ADSCA). METHOD: The family material and DNA samples were collected from thirteen families with ADSCA. To determine the characteristics of the CAG trinucleotide repeats in SCA1 gene, the PCR products of the Rep1 and Rep2 primers were analyzed, and the bands with CAG repeat expansion were cloned by PCR2. 1 vector and sequenced. RESULTS: One family was found to have an expanded CAG repeat in the 13 families with ADSCA. The clinically affected individual was heterozygous with one disease allele being 55 CAG repeats, whereas the mean size of the CAG repeats on 104 chromosomes generated from unrelated control Chinese individuals is 29.3 (ranging from 18 to 34). CONCLUSIONS: The frequency of the SCA1 mutation is about 7% in the 13 Chinese families with ADSCA, suggesting that this type of genetic defect is not the main cause involved in the pathogenesis of ADSCA in China. Since the mutation has also been found in Caucasian, Japanese, Malaysian, and Bangladeshi kindreds, it is suggested that this genetic defect may well have multiple origins in different ethnic groups. PMID- 10374380 TI - HLA-DQA1, -DQB1 polymorphism distribution in Chinese women with pregnancy induced hypertension in Shanghai area. AB - OBJECTIVE: To explore the association of human leukocyte antigen (HLA) with pregnancy induced hypertension (PIH). METHODS: We oligotyped HLA-DQA1, -DQB1 locus of 30 Chinese PIH families and 14 control families in Shanghai area by polymerase chain reaction-sequence specific oligonucleotide (PCR-SSO) hybridization method (probes labeled by nonradioactive technique). RESULTS: Compared with the control group, the allelic frequency of HLA-DQB1 * 0502 was significantly higher in PIH couples, and the sharing of HLA-DQA1 increased in PIH couples as well. No difference was found in HLA-DQA1 allelic frequencies or HLA DQB1 sharing between the two groups. Analysis of neither HLA-DQA1 nor HLA-DQB1 allelic frequencies in PIH patients and PIH mother-and-fetuses showed positive result. CONCLUSION: HLA-DQB1 * 0502 may be a marker of susceptibility to PIH. DQB1 * 0502 itself or some gene(s) located in HLA class II region and in linkage disequilibrium with 0502 affect maternal T cell immunity during pregnancy. The increase of compatibility in HLA-D region causes the production of blocking antibody to decrease. PMID- 10374381 TI - T-lymphocyte chemotaxis to IL-8 in patients with psoriasis in vitro. AB - OBJECTIVE: The critical role of infiltrating T cells in the pathogenesis of psoriasis is now well established. In order to determine whether circulating T cells from patients with psoriasis were also involved in the disease process, the authors investigated the biological behavior as studied by chemotactic activity of T cells in patients with psoriasis. METHODS: A 48 microchemotaxis chamber was employed to determine T-cell chemotaxis activity. In addition, the expression of T cell activation markers such as HLA-DR and interleukin 2 receptor were analysed with fluorescence activated cell sorting technique and serum IL-8 level was measured with ELISA methods. Forty-five patients with psoriasis (23 patients with severe psoriasis and 22 with mild psoriasis) and 21 patients with atopic dermatitis were investigated. For comparison, T-lymphocytes from 20 healthy controls were tested equally. RESULTS: T-cell chemotactic responses were significantly decreased in patients with severe psoriasis and atopic dermatitis as compared to healthy controls. Increased expression of activation markers such as HLA-DR and interleukin 2 receptor were demonstrated in circulating T cells from psoriatic patients and atopic dermatitis patients in comparison to healthy controls. Serum IL-8 level was significantly increased in patients with psoriasis and atopic dermatitis. CONCLUSIONS: Circulating T cells in patients with severe psoriasis show abnormal in vitro chemotactic response to IL-8. Furthermore, the in vivo activation state of T lymphocytes in these patients and increased level of serum IL-8 seemed to be associated to their decreased in vitro T-cell chemotactic responses. PMID- 10374383 TI - New concepts and advances of immobilization of long bones. AB - OBJECTIVE: To present some new concepts in the treatment of fractures and bone defects of long bones with internal fixation. METHODS: Animal experiments, mechanical tests and clinical analyses were done. RESULTS AND CONCLUSIONS: Reduction of fracture should be perfect, bone defect can be reconstructed by intramedullary and extramedullary bone graft. Relatively rigid fixation at the early stage and elastic fixation at the later stage are beneficial not only for fracture healing, but also for bone remodeling. In order to avoid complications including non-union, immobilization syndrome of the bone and joint, and implant failure, radiographs should be taken periodically; if there is any bone resorption, weight-bearing should be restricted. PMID- 10374382 TI - Employment of trauma and injury severity score and a severity characterization of trauma in the outcome evaluation of trauma care and their research advances. AB - OBJECTIVE: To review the application of trauma and injury severity score (TRISS) and a severity characterization of trauma (ASCOT) in the outcome evaluation of trauma care and their research advances. DATA SOURCES: Both Chinese- and English language literature searched by using MEDLINE/CD-ROM (1985-1996) and Index of Chinese-Language Literature (1985-1996). STUDY SELECTION: Over fifty papers and reviews published over the past ten years were selected. RESULTS AND CONCLUSION: TRISS can be employed for different purposes, that is, preliminary outcome-based evaluation (PRE) and definitive outcome-based evaluation (DEF). TRISS is a method which is now the most extensively used for the outcome evaluation of trauma. Even so, it still has some shortcomings, e.g., trauma can not be given the weights that should be given, and the section of age is too simple. ASCOT is also a physiologic and anatomic combined method for the evaluation of injury severity and outcome. To some extent, this method obviates the shortcomings of TRISS in the calculation of probability of survival (Ps) with injury severity score (ISS). Therefore, ASCOT is considered to be superior to TRISS in the evaluation of Ps. However, TRISS is still now more extensively used than ASCOT just because ASCOT was recently developed. PMID- 10374384 TI - Experimental and clinical studies of traumatic brain injury in China. PMID- 10374386 TI - One successful case of 6-organ-failure after severe polytrauma. PMID- 10374387 TI - In situ saphenous vein arterial bypass for lower limb ischemia. PMID- 10374385 TI - Detection of hepatocellular carcinoma cells in peripheral venous blood and its significance. PMID- 10374388 TI - Expression of estrogen receptors and progesterone receptors in transitional cell carcinoma of bladder. PMID- 10374389 TI - Starting thrombolytic therapy for patients with acute myocardial infarction in Accident and Emergency Department: from implementation to evaluation. AB - OBJECTIVE: To evaluate the effectiveness of initiating thrombolysis for patients with acute myocardial infarction (AMI) in the Accident and Emergency Department. METHODS: From January 1993 to December 1995, all AMI patients who were admitted to the United Christian Hospital and given thrombolytic therapy were studied. The patients' demographic data, time and mode of presentation, site of myocardial infarction, treatment modality and timing, and complications related to AMI or treatment were recorded prospectively in our AMI database. The frequency of thrombolysis administered in Accident and Emergency Department and Coronary Care Unit, as well as the median door-to-needle time (time interval between hospital arrival to initiation of thrombolytic therapy) were compared. Cases of inappropriate thrombolysis and complication were also analyzed. RESULTS: Over these 3 years, 257 patients received thrombolysis in the United Christian Hospital. The percentage of patients receiving thrombolysis in Accident and Emergency Department increased from 3.2% in 1993 to 12.3% in 1994, and to 39.4% in 1995. The median time interval between arrival to hospital and thrombolysis (door-to-needle time) was 25 minutes, compared with 81 minutes in the Coronary Care Unit. The door-to-needle time also improved over these 3 years: from 95 minutes in 1993 to 75 minutes in 1995 in Coronary Care Unit group, and from 35 minutes in 1993 to 20 minutes in 1995 in the Accident and Emergency Department group. Over these 3 years, 2 cases of inappropriate thrombolysis were reported but these did not result in any mortality. Four complications from thrombolytic therapy were reported, and these were managed appropriately by the staff in Accident and Emergency Department and did not result in mortality. CONCLUSIONS: Starting thrombolytic therapy in Accident and Emergency Department is safe and effectively decreases the door-to-needle time. PMID- 10374390 TI - Molecular level investigation of exocytosis in cultured human pheochromocytoma cells: insights from high resolution scanning electron microscopy combined with autoradiogram and cytochemistry. AB - OBJECTIVE: To investigate molecular events of exocytosis in cultured human pheochromocytoma cells with stimulation. METHODS: The cultured pheochromocytoma cells prepared from human adrenal pheochromocytoma tumor were stimulated for the release of catecholamines by depolarization with the administration of 50 mmol/L KCl. Transmission electron microscopy (TEM) and high resolution scanning electron microscopy (HR-SEM) combined with autoradiography and cytochemistry were used to observe molecular mechanisms of exocytotic release of catecholamines from the stimulated cells labelled with 3H-noradrenaline and the filipin-treated cells. RESULTS: TEM and HR-SEM observations of the stimulated cells labelled with 3H noradrenaline revealed that the initial exocytotic fusion pores even less than 10 nm in diameter in human pheochromocytoma cells can be clearly observed in a single lipid bilayer. Furthermore, HR-SEM examinations of the filipin-treated cells showed that the derangement of the particles of the filipin-sterol complexes (FSCs) in the fused membranes of granule and plasma membranes occurred as the exocytotic fusion pores opened. In addition, the aggreates of the FSCs particles were consistently demonstrated around the openings of the differently sized closing exocytotic pores. CONCLUSIONS: Based on our results, it is suggested that the rearrangement of the sterol molecules in the fused membranes of granule and plasma membranes plays an important role in the opening and closing mechanisms of exocytotic fusion pores. We hope that morphological data obtained in this study can provide some new insights into the understanding of molecular mechanisms of exocytosis, particularly the opening and closing of exocytotic fusion pores in relation to the distribution of the membrane sterols. PMID- 10374391 TI - Minimally invasive coronary surgery in women. AB - OBJECTIVE: To evaluate the minimally invasive surgery in coronary artery bypass grafting and the feasibility for revascularization of triple vessel coronary artery disease. METHODS: Nine female patients, aged 49.1 to 81.6 years (mean 64.3), were operated on for triple vessel disease through minimally invasive surgical techniques. The surgeries were performed through limited left parasternal incision under femorofemoral extracorporeal circulation. The myocardium was protected by antegrade infusion of cold blood cardioplegic solution while the aorta was cross-clamped. Under direct vision, the left saphenous vein grafts were connected sequentially to the diagonal branch, obtuse marginal branch and posterior descending branch, and the left internal thoracic arterial graft was anastomosed to the left anterior descending artery in each patient. RESULTS: The number of distal anastomoses was 3 to 4 with a mean of 3.7. The aortic crossclamp time was 52 to 130 minutes (82 +/- 25 minutes). The duration of extracorporeal circulation was 78 to 151 minutes (115 +/- 29 minutes). The postoperative course was uneventful in all patients. The postoperative length of stay was 4 to 12 days (7.2 +/- 2.0 days). Follow-up (4.2 to 8.7 months, mean 6.4) was complete in all patients and there were no late deaths or angina. Coronary angiography of 2 patients showed patent grafts. All patients were satisfied with the good cosmetic healing of the incision. CONCLUSION: Our experience demonstrates that minimally invasive surgery in coronary artery bypass grafting is technically feasible and may be an alternative approach in surgical revascularization of triple vessel coronary artery disease, especially in female patients. PMID- 10374392 TI - Protective effect of the angiotensin-converting enzyme inhibitor perindopril on diabetic glomerulopathy in streptozotocin-induced diabetic rats. AB - OBJECTIVES: To evaluate the protective effect of the angiotensin-converting enzyme inhibitor perindopril on diabetic glomerulopathy in rats with experimentally induced diabetes and explore its possible mechanisms. METHODS: Ninety-two adult male Wistar rats were randomly allocated into diabetes mellitus (DM), diabetes mellitus + perindopril (DMP) and control (C) groups. According to the duration of diabetes or observation (1, 3, 6 months), each group was randomly subdivided into DM1, DM3, DM6; DMP1, DMP3, DMP6; and C1, C3, C6 groups. Diabetes was induced by intraperitoneal injection of streptozotocin. The rats in the DMP groups received perindopril 1 mg.kg-1.d-1, through gastric intubation. Urinary protein excretion rate was determined by the method of Coomassie brilliant blue. Plasma renin activity, renal tissue renin activity, and plasma and renal tissue angiotensin II concentration were assayed by radioimmunoassay (RIA). Renal tissue total RNA was extracted by the Chomezymskis AGPC method. Renal angiotensinogen mRNA expression level was assessed by slot blot hybridization using a full length rat angiotensinogen cDNA probe labelled with 32P-dCTP and a random primer. RESULTS: There was increased activity of the renin angiotensin system in diabetic rats. Perindopril decreased proteinuria and delayed the progression of glomerular basement membrane thickening. However, it did not reduce the expansion of the mesangial matrix (P < 0.05). Renin activity increased and angiotensin II concentration decreased significantly in both plasma and renal tissue in diabetes + perindopril groups (P < 0.05). CONCLUSIONS: Perindopril may help prevent the progression of diabetic glomerulopathy, and the inhibition of renin angiotensin system activity may be a mechanism for this action. PMID- 10374393 TI - The pathogenetic role of endogenous angiotensin II in stress ulcer in obstructive jaundice rats. AB - OBJECTIVE: To investigate the pathogenetic role of endogenous angiotensin II (Ang II) in the mechanism of stress ulcer in obstructive jaundice rats and to detect the effect of angiotensin converting enzyme inhibitor (ACEI) on stress ulcer in obstructive jaundice rats. METHODS: After common bile duct ligation (CBDL) in Wistar rats, the content of plasma and gastric mucosal Ang II, gastric mucosal blood flow (GMBF) and gastric mucosal damage were measured, and the relationship among them was analyzed. RESULTS: The plasma Ang II contents increased much more significantly at 1, 3, 7 and 14 days following CBDL than those in non-CBDL rats (P < 0.05, < 0.01, < 0.01 and < 0.01, respectively). Within 120 minutes following cold-restraint stress, plasma and gastric mucosal Ang II contents were elevated, GMBF decreased, and ulcer index and gastric mucosal damage increased more significantly than those in non-cold-restraint stress rats (P < 0.05, < 0.05, < 0.01, < 0.01 and < 0.05, respectively). Administration of an ACEI, enalaprili, to CBDL rats (5 mg.kg-1.day-1, orally for two days) before stress reduced both the plasma and gastric mucosal Ang II levels, inhibited the decrease of GMBF and decreased ulcer index and gastric mucosal damage (P < 0.001, < 0.01, < 0.01, < 0.01 and < 0.05, respectively). CONCLUSION: The endogenous Ang II plays a significant pathogenetic role in the development of stress ulcer in obstructive jaundice rats, and ACEI may prevent stress ulcer. PMID- 10374394 TI - Alterations of oncogenes, tumor suppressor genes and growth factors in hepatocellular carcinoma: with relation to tumor size and invasiveness. AB - OBJECTIVE: To make a better understanding of the molecular mechanisms involved in recurrence and metastasis of the hepatocellular carcinoma (HCC), some invasion related oncogenes, and growth factors have been investigated. METHODS: The studies were separately carried out, the results of which were summarized in this article with relation to tumor size and invasiveness of HCC. RESULTS: The aberration rates of p53 and CDKN2 in HCC were 45.9% and 36.4% respectively, which were higher in invasive HCC compared with non-invasive HCC. H-ras expression was positive in 29.3% of HCC, which was associated with recurrence and extrahepatic metastasis of HCC. Intralesional injection of H-ras antisense gene markedly inhibited the tumor growth and metastasis of HCC in nude mice. The positive rates of transforming growth factor (TGF)-alpha, epidermal growth factor receptor (EGFR) and c-erbB-2 were 45.7%, 47.1% and 92.3% respectively. The expression of EGFR was closely related to TGF-alpha, which was related to HCC recurrence. But no obvious difference of TGF-alpha or c-erbB-2 expression was found between HCC with and without recurrence, or with and without extrahepatic metastasis. Expression of nm23/tissue inhibitor of metalloproteinase (TIMP)-2 was positively associated with the prognosis of HCC patients (Log-rank, P < 0.001). The alterative rates of above-mentioned genes and growth factors in small HCC were slightly lower than that in large ones, but no significant difference was shown except the p53 mutation. CONCLUSIONS: The p53/CDKN2 mutation, over-expression of H-ras/EGFR, were associated with the invasiveness and recurrence of HCC. H-ras antisense gene might be of potential implication in the control of HCC recurrence and metastasis. Expression of nm23/TIMP-2 was closely related to the prognosis of HCC patients. Biological characteristics remained critical points to the prognosis even in small HCC. PMID- 10374395 TI - Enhanced expression of HLA class I molecules in human hepatocellular carcinoma cells transduced with gamma-interferon gene. AB - OBJECTIVE: To investigate the expression of exogenous gamma-interferon gene in human hepatocellular carcinoma cells following retroviral transduction and the effect on the expression of surface HLA class I molecules. METHODS: Retroviral vector pLXSN was used to introduce human gamma-interferon (IFN-gamma) gene into four different human hepatocellular carcinoma cell lines (HCC). The G418 resistant colonies were isolated and cloned. The integration and expression of IFN-gamma gene were determined by PCR and RT-PCR analysis. A bioassay method was used to test the amount of IFN-gamma secreted by gene modified HCC cells. The expression of HLA class I molecules in HCC cells were analyzed by flow cytometry using indirect fluorescence staining. RESULTS: Four different HCC cell lines were successfully transduced with human IFN-gamma gene using retroviral vector. The integration and expression of IFN-gamma gene were shown only in the transduced cells. All four genetically modified HCC cells can secrete varied amount of IFN gamma and demonstrate a significant up-regulation of surface HLA class I antigens. One specific HLA class I antigen, HLA-A2, has almost the same degree of increase as that of the total HLA class I molecules after transduction with IFN gamma gene. CONCLUSIONS: Gene modification with IFN-gamma gene can significantly enhance the expression of HLA class I molecules in HCC cells and may increase its immunogenicity. These gene modified tumor vaccines can be helpful in tumor biotherapy. PMID- 10374396 TI - The role of endothelin-1 in the aorta of post-infarct left ventricular dysfunction rats treated with captopril. AB - OBJECTIVES: There is little information available regarding local vasomotor regulating processes in chronic heart failure. In this study, we tested the hypothesis that chronic heart failure impaired the endothelial function, and long term captopril treatment might reverse endothelial activity through tissue endothelin (ET) pathway. METHODS: Forty Sprague-Dawley rats were divided into 4 groups including 15 rats in each of the sham-operated with or without captopril treated groups and 5 rats in each of large infarcted with or without captopril treated groups. RESULTS: Concentration-response curves obtained in aortic rings without endothelium revealed no difference in nitroprusside-induced relaxation. With endothelium, rightward shifting was noted only in the untreated large infarct group during acetylcholine-induced relaxation. As compared to the non treated group, plasma ET-1 concentrations were lower in the captopril-treated with or without large infarct groups. However, endothelin-like immunoreactivity in endothelial cells and cytoplasma of smooth muscle cells of the media of the aorta were lower only in the non-treated large infarct group. CONCLUSIONS: Endothelial function was impaired in the chronic heart failure model. Coverting enzyme inhibitor might improve endothelial function through the Local endothelin pathway. PMID- 10374397 TI - Endothelium-dependent relaxation of canine pulmonary artery endothelium after prolonged preservation. AB - OBJECTIVE: Experiments were designed to investigate the effect of Euro-Collins (EC) solution and University of Wisconsin (UW) solution on function of pulmonary arterial endothelium. METHODS: Third order canine pulmonary artery segments were preserved in cold (4 degrees C) UW (group 1, n = 8) or EC (group 2, n = 9) solutions for 16 hours. The preserved (group 1 and 2) and control (group 3, n = 7) pulmonary arterial segments with and without endothelium were studied in vitro in organ chambers to measure isometric tension. RESULTS: The endothelium dependent relaxation to acetylcholine and adenosine diphosphate of group 1 and 3 were significantly better than those of group 2. CONCLUSIONS: We concluded that endothelium-dependent relaxation of canine pulmonary arterial endothelium to receptor-dependent acetylcholine and adenosine diphosphate were impaired after preservation with Euro-Collins solution. However, endothelium-dependent relaxation of pulmonary segments were well maintained after preservation with University of Wisconsin solution. PMID- 10374398 TI - Influence of recombinant retroviral vector expressing antisense TGF alpha on malignant phenotype of human pancreatic carcinoma cell line. AB - OBJECTIVE: To observe the effects of antisense TGF alpha on the growth of human pancreatic carcinoma cell line cells. METHODS: A recombinant retroviral vector expressing antisense TGF alpha was constructed, and transfected the ecotropic packaging cell line psi-2 with lipofectin. After the amphotropic packaging cell line PA317 was transfected with the virus supernatant of psi-2, the replication defective, amphotropic retroviral supernatant was used to infect human pancreatic carcinoma cell line PC-7. Following puromycin selection, puromycin-resistant colonies were pooled and expanded to a cell line PC-7/AS-TGF alpha. RESULTS: The retroviral integration in the genomes of psi-2, PA317 and transformant PC-7 cells was confirmed by Southern blot hybridization. Northern blot hybridization showed a down regulation of endogenous TGF alpha in PC-7/AS-TGF alpha cell line. The high levels of growth inhibition and reduction of 3H-TdR incorporation in PC-7/AS TGF alpha were evident. Also, the soft agar colony-formation and tumorigenicity in nude mice were significantly suppressed by antisense TGF alpha. CONCLUSION: The antisense TGF alpha expressing vector can block the target gene expression, suppress the cell growth and partially reverse the malignant phenotype of pancreatic carcinoma cells. PMID- 10374400 TI - N-ras mutations in 43 Chinese cases of acute myeloid leukemia. AB - OBJECTIVE: To detect 3 kinds of N-ras mutations in Chinese patients with acute myeloid leukemia (AML). METHODS: In vitro DNA amplification followed by oligonucleotide dot analysis were used to study N-ras gene mutations in 43 cases of acute myeloid leukemia (AML). 25 healthy people were used as controls. Patients were selected in the Beijing district and consisted of 19 males and 24 females. The average age was 37. The controls were healthy individuals with the average age of 36.5 from the same region. 3 oligonucleotide probes were artificially synthesized to detect mutations in codon 12 and 13 of N-ras. RESULTS: Five out of 43 AML samples have been found contain G-->A mutation in codon 12.2 have G-->T mutation in codon 12. One has G-->A mutation in codon 13. The mutation rate was 18.6%. None of the controls presented these mutations. The frequency of mutation of N-ras in the AML samples showed statistical differences with that of the controls. CONCLUSION: Analysis of the results suggests the N-ras mutations may have some relationship with the etiology of acute myeloid leukemia. PMID- 10374399 TI - Detection of p53 gene mutations in human leukemia by PCR-SSCP analysis and direct nucleotide sequencing. AB - OBJECTIVE: To look for mutations of the p53 gene in leukemic patients and study the relationship between abnormalities in p53 gene and leukemogenesis. METHODS: The peripheral blood and Bone Marrow Samples were collected from 36 patients with various leukemia types including 14 cases of lymphocytic leukemia [8 cases of acute lymphocytic leukemia (ALL), 4 cases of chronic lymphocytic leukemia (CLL), 2 cases of hairy cell leukemia (HCL)] and 22 cases of myelocytic leukemia [11 cases of acute nonlymphocytic leukemia (ANLL), 11 cases of chronic myelocytic leukemia (CML)]. DNA structures of exon 5-8 of the p53 gene were scanned by PCR SSCP (single strand conformation polymorphism analysis of polymerase chain reaction products). The appropriate DNA fragments were amplified, Purified and sequenced directly. RESULTS: By PCR-SSCP analysis, shifts in electrophoretic mobility of the p53 gene were detected in 3 of 14 patients with lymphocytic leukemia (2 ALL and 1 CLL), but none in 22 patients with myelocytic leukemia including one in blastic crisis. Direct nucleotide sequencing in one patient with ALL showed transition of CTG to CAG at codon 257 of exon 7, resulting in a change of its encoded amino acids from aspartic acid to valine. To our knowledge, the mutation at this codon has not been previously reported hitherto. CONCLUSIONS: The p53 gene mutations are specifically associated with lymphocytic leukemia. Alternations of the p53 gene may play a certain role in leukemogenesis in some cases of lymphocytic leukemia. PMID- 10374401 TI - Diagnosis and classification of hepatic echinococcosis by ultrasonography. AB - OBJECTIVE: To make an early and correct diagnosis of hepatic echinococcosis. METHODS: A total of 1092 patients with hepatic echinococcosis underwent operation in our hospital between 1984 and 1995. Of these patients, 427 (39.1%) were cases with complications, including secondary infection, rupture, obstructive jaundice, anaphylactic shock, disseminated implantation with resultant multiple echinococcosis and portal hypertension. The ultrasonic examination has been generally used in clinical practice for comprehensive evaluation which can dectedct the preoperative diagnostic accuracy rate. RESULTS: B-mode ultrasonography can be used not only to detect the location, dimension and pathological characteristics of hydatid cyst but also to show the pathological changes of various complications caused by echinococcosis. Seven patterns specific ultrasonic scanning images were revealed. In this series the diagnostic accuracy rate reached 98.8%. CONCLUSIONS: Ultrasonic examination is harmless to human body, and has been widely used in combination with immunological tests in clinical and epidemiological studies to detect the asymptomatic parasite carriers in early stage and to improve the preoperative diagnostic accuracy rate. PMID- 10374402 TI - Diagnosis and treatment of acute central cervical cord injury. AB - OBJECTIVE: To clarify the diagnosis and management of acute central cervical cord injury. METHODS: Eighty-nine patients with acute cervical central cord injury were retrospectively reviewed. Sixty-three patients were treated conservatively and 26 were treated surgically. There were two acute deaths. Eighty-seven patients were followed up for 3 months to 15 years. RESULTS: Their average neurological score (ASIA) was increased from 41.7 at admission to 83.1 at follow up. CONCLUSIONS: Acute central cervical cord injury should be differentiated from complete spinal cord injury, cervical myelopathy, cruciate paralysis and C8 nerve root injury. When compression of nerve tissue or cervical instability is identified, operative intervention should be indicated. The prognosis is less optimistic in the patients with severe primary injury and at old age. PMID- 10374403 TI - Inner ear damage in guinea pigs exposed to stable and impulse noise. AB - OBJECTIVE: To investigate the inner ear damage after exposure to stable noise, impulse noise and stable plus impulse noise in guinea pigs. METHODS: Ninety-six healthy guinea pigs were divided into 3 equal groups. (1) Stable noise group: exposed to 110 dBA stable noise for 3 days, 4 hours per day. (2) Impulse noise group: exposed to 165 dBA simulated cannon fire impulse noise 10 times successively at an interval of 10 seconds. (3) stable plus impulse noise group: exposed to the same stable noise as that in the first group, then after a 2-hour rest, the animals were followed with impulse noise exposures as that in the second group. After those exposure, each of the 3 groups was further divided into 4 subgroups according to the time after the noise exposure, namely, the right after, 7 d, 14 d and 30 d groups. The evoked cortical potential responses to click and tone burst stimulation sound were examined. The surface preparation and celloidine embedded serial section of the cochlea were observed under a light microscope. RESULTS: Both the stable and impulse noise could increase the hearing threshold and damage the inner ear hair cells. The damage in the first group was relatively slight, whereas in group 3 the damage was more severe than that in the other 2 groups. CONCLUSION: For seamen who are working in heavy noise environment, corresponding measures should be taken to protect their ears from noise which induces hearing loss. PMID- 10374404 TI - Enhancement ablation for the treatment of undercorrection after excimer laser in situ keratomileusis for correcting myopia. AB - OBJECTIVE: To evaluate the treatment of undercorrection after the excimer laser in situ keratomileusis (LASIK) for correcting moderate and high myopia. METHODS: An enhancement ablation was performed in 48 eyes of 39 patients who had undergone LASIK but remained in undercorrection. Four procedures were performed within 1 month postoperatively, and the others performed between 3 and 10 months. The surgical technique includes the re-invert of the corneal cap from the temporal side, the excimer laser ablation, and the re-position of the cap. RESULTS: The undercorrection (spherical equivalent) ranged from -2.00 to -11.00 D, with a mean of -4.34D +/- 1.95 D. Following up after enhancement ablation was done after 4 to 12 months, the refractions in the 42 eyes were found to be within +/- 1.00 D. Undercorrection of -2.50 D to -5.00 D recurred in 6 eyes. Uncorrected visual acuity equals to the preoperative spectacle corrected visual acuity in 39 of 48 eyes (81.3%). Five eyes gained 1 line, 1 eye gained 2 lines and 4 eyes lost 1 line. No eyes had haze. CONCLUSION: Undercorrection after LASIK can be corrected by an enhancement ablation of the stroma under the primary corneal cap with a 193 nm ArF excimer laser, and the time for the enhancement of ablation is at 3 months postoperatively. PMID- 10374406 TI - Sex hormones are weak regulators of HPV16 DNA-immortalized human uterine exocervical epithelial cells. AB - OBJECTIVE: To examine the effect of sex hormones on the growth and gene expression of human papillomavirus (HPV) type 16 DNA-immortalized human uterine cervical epithelial cells (HCE16/3 cells). METHODS: The effect of sex hormones on the growth and viral gene expression of HCE16/3 cells was studied using [3H] thymidine incorporation, soft agarose assay and Northern blot analysis. RESULTS: The growth of HCE16/3 cells was found to be little affected by estradiol and progesterone while insulin was a mitogen for HCE16/3 cells in phenol redfree medium with steroid-stripped serum. Furthermore, synergistic effects between insulin and hormones were not observed. Estradiol could not induce the growth of HCE16/3 cell line in soft agarose, either. In Northern blot analysis, however, the hormones upregulated the HPV 16 early gene expression in HCE16/3 cells, which was generally considered to be required for the proliferation of HPV DNA immortalized cells. CONCLUSIONS: These results suggest that the proliferation of HCE16/3 cells is still dependent on growth factors and sex hormones upregulate the HPV 16 early gene expression. More researches should be done before elucidating the effects of sex hormones on human cervical cancinogenesis. PMID- 10374405 TI - CT-guided stereotactic biopsy of deep brain lesions: report of 310 cases. AB - OBJECTIVE: To evaluate the accuracy of CT-guided stereotactic biopsy in making correct pathological diagnosis and choosing corresponding management of brain tumors. METHODS: From 1991 to 1995, CT-guided stereotactic biopsy was performed in 310 patients with intracerebral lesions which were deep-seated or located in certain main functional areas. The patients were 198 men and 112 women. Their ages ranged from 4.5 to 70 years (average: 39.3 years). The lesions were located in the deep cerebrum (74 patients), the sellar area (62), the basal ganglion (51), the posterior part of the third ventricle (38), other intraventricleular area (21), the cerebellum (17) and the brain stem (9), and intracranial multiple lesions were found in 38 patients. RESULTS: Brain tumors were diagnosed pathologically in 266 patients (85.8%); inflammatory process in 25 (8.1%), other lesions in 8 (2.6%) and uncertain cases were 11 (3.6%). The overall positive rate of biopsy was 96.4% and the positive rate for brain tumor was 85.8%. Intracranial hematomas after biopsy were found in 5 patients (1.6%). There were no deaths induced by the biopsy or other serious complications. CONCLUSIONS: The results suggest that CT-guided stereotactic biopsy is a reliable method for histopathological diagnosis of brain tumors and it is also of great help in selecting appropriate management. PMID- 10374407 TI - Effect of mild hypothermia on the changes of cerebral blood flow, brain blood barrier and neuronal injuries following reperfusion of focal cerebral ischemia in rats. AB - OBJECTIVE: To compare the effects of mild hypothermia induced in different time courses on rats subjected to 3 hours (h) of ischemia followed by 3 h or 72 h of reperfusion. METHODS: Eighty male Sprague-Dawley rats were divided into three mild hypothermic (MHT, 32 +/- 0.2 degrees C) groups, including intra-ischemia (MHTi), intra-reperfusion (MHTr), and intra-ischemia/reperfusion (MHTi + r) group, and one normothermic group (NT, 37 +/- 0.2 degrees C) as the control. Reversible focal ischemia was carried out in rats with suture model. The cortical blood flow was measured during 3 h of ischemia followed by 3 h of reperfusion. The permeability of brain blood barrier (BBB) was estimated after 3 h of reperfusion. The infarct volume was measured at 72 h after reperfusion to determine the effects of MHT. RESULTS: The acute post-ischemic hyperperfusion and delayed hypoperfusion in ischemic perifocal region and sustained hypoperfusion in ischemic core were inhibited in MHTi + r and MHTi rats (P < 0.05). MHTi + r protection on post-ischemic progressive hypoperfusion in the perifocal region was more effective than that of MHTi (P < 0.05). The BBB disruption and the infarct volume were significantly reduced in both MHTi and MHTi + r groups (P < 0.05), especially in the MHTi + r rats. CONCLUSIONS: This study demonstrates that MHTi + r has more substantial protective effects on reducing ischemia/reperfusion injury than MHTi. It may inhibit post-ischemic hyperperfusion and delayed or sustained hypoperfusion in ischemic perifocal regions, and reduce brain blood barrier disruption in the cortex region. PMID- 10374408 TI - Cellular composition and anatomic distribution in nonfunctioning pancreatic endocrine tumors: immunohistochemical study of 30 cases. AB - OBJECTIVE: To investigate the cytological pattern and distribution in nonfunctioning pancreatic endocrine tumors. METHODS: Using labeled streptavidin biotin (LSAB), immunohistochemical staining for insulin, glucagon, somatostatin, pancreatic polypeptide and gastrin was performed on 30 nonfunctioning pancreatic endocrine tumors from 30 patients. The cellular composition and anatomic distribution in these tumors were analyzed. RESULTS: Of 30 tumor tissues, 22 (73.3%) were found to contain cells immunoreactive to 1-4 kinds of peptide hormones; 17 (56.7%) showed positive staining for more than one peptide and up to 4 peptides; and 8 (26.7%) showed negative immunoreaction to all antiserum applied. No tumor was found to contain immunoreactive gastrin. Among 17 multihormonal tumors, 4 contained 2 kinds of peptide hormones, 8 had 3 kinds, and 5 harbored 4 kinds of peptide hormones. In addition, the difference in the number and type of positive endocrine cells between the tumors arising from the head of the pancreas and those arising from the body and tail of the pancreas were statistically significant (P < 0.05). CONCLUSIONS: Immunohistochemically, the high positive rate to peptide hormones suggests that the nonfunctioning pancreatic endocrine tumors are actually not nonfunctioning; they are asymptomatic pancreatic endocrine tumors. Moreover, an uneven distribution of positive endocrine cells in the nonfunctioning pancreas endocrine tumors within the pancreas was identified. PMID- 10374409 TI - Ewing's sarcoma of the maxilla. PMID- 10374411 TI - A preliminary clinical study of bifidobacteria preparation on the treatment of diarrhea in severely burned patients. AB - OBJECTIVE: To investigate the role of bifidobacteria preparation in the prevention of diarrhea in severely burned patients. METHODS: Forty-three severely burned patients who were inflicted by diarrhea were included in this study. The changes of intestinal microflora were observed. RESULTS: The intestinal flora of all patients with diarrhea changed greatly before treatment. The proportions of bifidobacteria and bacteroid decreased significantly, whereas those of aerobe and Candida increased relatively. The ratio of anaerobe to aerobe decreased. As a result, the patients' intestinal flora were distorted. After six days treatment of bifidobacteria, diarrhea in most patients ceased and the intestinal microflora restored. CONCLUSION: Bifidobacteria feeding plays an important role in restoring intestinal microflora and stopping diarrhea in severely burned patients. PMID- 10374410 TI - p53 expression in human gestational trophoblastic tumors. PMID- 10374412 TI - Cloning and expression of a novel partial cDNA related to hypertension. PMID- 10374413 TI - A resume of epidemiological surveys of several main rheumatic diseases in China. PMID- 10374414 TI - Melatonin: a chemical photoperiodic signal with clinical significance in humans. AB - Secretion of pineal melatonin exhibits a diumal rhythm and a seasonal rhythm in humans. Night-time melatonin is high at 3-5 year-old and decreases with age. Many drugs and pathological conditions also change melatonin levels in the circulation. Melatonin has a mild sedative effect and has been used effectively in synchronizing the sleep-wake cycle of patients with sleep disorders. Immunoenhancing, anti-cancer, anti-aging and anti-oxidant effects of melatonin have been proposed. Recent studies suggest that melatonin receptors are present in central and peripheral tissues. The importance of melatonin receptors on the nervous, reproductive, immune and renal functions is implicated. Studies on the molecular biology, physiology and pathology of melatonin receptors in different tissues are progressing rapidly. The physiological and pathological changes in melatonin secretion, multifarious melatonin actions, and diverse melatonin receptors reported suggest that melatonin is a photoperiodic signal with clinical significance in humans. PMID- 10374415 TI - Anti-Sa antibody in Chinese rheumatoid arthritis. AB - OBJECTIVE: To test anti-Sa antibody in different autoimmune connective tissue diseases and analyze the relationship between Sa antibody and clinical manifestations and laboratory tests in rheumatoid arthritis. METHOD: Sa antigen was extracted from human placenta. Anti-Sa antibody was tested in 40 normal people and 478 connective tissue disease (CTD) patients using Western Blotting (WB). RESULTS: Sa antigen was a protein with molecular weights of 50 kD and 55 kD. Anti-Sa antibody was positive in 31.9% (61/191) rheumatoid arthritis (RA), 3.0% (2/67) Sjogren's syndrome (SS), 4.3% (2/46) systemic lupus erythmatosus (SLE) and 0% (0/66) Behcet's disease, 0% (0/60) polymyositis/dermatomyositis (PM/DM), 0% (0/66) other CTD and 0% (0/40) normal controls. Anti-Sa antibody was different from other auto-antibodies in RA. In rheumatoid arthritis its sensitivity, specificity, positive prediction rate, negative prediction rate were 31.9%, 98.6%, 93.8% and 68.5% respectively. Anti-Sa antibody positive patients were significantly different from anti-Sa antibody negative patients in moming stiffness, ESR, ANA and X-ray grade. CONCLUSION: Anti-Sa antibody was a new auto antibody for the diagnosis of RA. Anti-Sa antibody positive patients seem to have more serious inflammation and more advanced disease process. PMID- 10374416 TI - HLA-DRB1 genes in 5 rheumatic disease multi-case families. AB - OBJECTIVE: To detect HLA-DRB1 (DR1-10) alleles in 5 families with multi-case rheumatic diseases, and to study the possible influence of DRB1 genes in the pathogenesis of rheumatic diseases. METHODS: Sequence-Specific Primer PCR (PCR SSP) method was used to examine HLA-DRB1 alleles. Totally 36 members of 5 families and 166 healthy people were involved in this study. The results were assessed by Chi-square test. RESULTS: The HLA-DRB1 allele frequency in the patients and their relatives was similar. No significant difference was found. But DR4 allele frequency in the patients (90.9%) and their relatives (68%) was much higher than that in normal controls (16.8%) and the difference was statistically significant (P < 0.0001). In family 4, two RA patients have different DRB1 alleles, while in family 5, two patients have the same DRB1 alleles, one developed SLE and the other developed RA. CONCLUSIONS: DR4 is closely related to rheumatoid arthritis. The nelatives of RA patients may be at greater risk to develop RA than individuals without family history. Some patients had the same DRB1 allele but developed different rheumatic diseases. This suggested that there might be some common pathways in genetic predisposing of rheumatic diseases. On the other hand, only a few patients with the same DRB1 allele developed rheumatic diseases during their life, so other factors besides DRB1 gene might also be involved in the pathogenesis of rheumatic diseases. PMID- 10374417 TI - Immunohistochemical study of Fas antigen expression in synovial tissues from patients with rheumatoid arthritis. AB - OBJECTIVE: To investigate the location and expression of Fas (Apo-1, CD-95) antigen in synovial tissue from rheumatoid arthritis (RA). METHODS: Immunohistochemical technique was used to identify the location and expression of Fas antigen in synovial tissues from 27 RA, 11 osteoarthritis (OA), 7 ankloysing spondylitis (AS), 3 pigmented villo-hyperplastic synovitis, 1 juvenile rheumatoid (JRA) patients and 5 "normal" control subjects. RESULTS: Fas was strongly expressed by synoviocytes and infiltrated lymphocytes in approximately two-thirds of RA patients (16/27). However, only weak expression occurred on lymphocytes in 3 of 11 OA, 1 of 7 AS patients and 1 of 5 "normal" subjects. The stain-positive substance in the forms of rings, granules, or dust was deposited on the cell membrane and in cytoplasm. CONCLUSION: The expression of Fas may be involved in the mechanism of synovium proliferation and abnormal activation of local lymphocytes in RA. PMID- 10374418 TI - Mixed connective tissue disease: a disease entity? AB - OBJECTIVE: To explicate whether mixed connective tissue disease (MCTD) is a distinct disease and evaluate the reliability of three different diagnostic criteria proposed by Sharp, Alarcon-Segovia and Kasukawa respectively. METHODS: Clinical follow-up of 50 MCTD patients lasted 2-8 years (80% > 5 years). HLA-A, B as well as -DR typing was performed by complemently dependent cytotocity assay. Autoantibody profile was detected by counterimmune electrophoresis (CIE). RESULTS: Thirteen (26.0%) of the 50 MCTD patients subsequently developed other connective tissue disease (OCTD), including 7 systemic lupus erythematosis (SLE), and 6 progressive systemic scleroderma (PSS). Among 23 of the MCTD patients fulfilling Sharp's criteria, 1 (4.3%) developed PSS, but among 23 of the patients fulfilling Kasukawa's, not Sharp's, 7 (30.4%) developed OCTD and among 27 of the patients fulfilling Alarcon-Segovia's, not Sharp's, 12 (44.4%) developed OCTD. In the frequencies of DR4 and DR5, there were significant differences between patients fulfilling Sharp's (60.9%, 56.5%) and controls (24.3%, P < 0.005, RR = 4.7 and 21.4%, P < 0.005, RR = 4.6%), but there were no significant differences between the patients not fulfilling Sharp's and normal control (P > 0.05). CONCLUSIONS: MCTD is a distict rheumatic disease. Sharp's criteria is the most reliable for diagnosis of MCTD. PMID- 10374419 TI - Primary Sjogren's syndrome and its lymphoid malignancy: a report of four cases. AB - OBJECTIVE: To evaluate the incidence and spectrum of malignancy of primary Sjogren's syndrome (pSS). METHODS: 250 pSS who were followed-up in Peking Union Medical College (PUMC) Hospital were analyzed. RESULTS: Four of them were diagnosed with histopathological findings of 2 non-Hodgkin Lymphoma, 1 AILD, 1 multiple myeloma. Two died of secondary infection while receiving chemotherapy for lymphoma, 2 remained remitted. CONCLUSIONS: The risk factors were persistent enlargement of major salivary glands, appearance of monoclonal serum lg, and disappearance of auto antibodies. PMID- 10374420 TI - Clinical manifestations and immunological features of primary Sjogren's syndrome with liver involvement: analysis of thirty cases. AB - OBJECTIVE: To evaluate the incidence, severity, clinical manifestations and immunological features relevant to liver involvement in 135 cases of primary Sjogren's syndrome. METHODS: One hundred and thirty-five patients with definite primary Sjogren's syndrome were analyzed retrospectively for liver involvement by the abnormalities of the liver enzymes, bilirubin level and liver biopsied section. RESULTS: The liver involvement in 30 patients (22.2%) could be etiologically ascribed to Sjogren's syndrome itself. The clinical spectrum and severity of this entity differed widely, 36.6% showed no relevant clinical symptoms, however jaundice was found in 46.7% of patients. Six patients showed pathological changes of chronic active hepatitis. 73.3% of all patients with liver involvement responded to steroid and immunosuppressive drugs, yet with a tendency to relapse (two cases). Liver cirrhosis was developed in five cases. The spectrum of serum autoantibodies in the patients with liver involvement showed no difference from those without liver involvement. Most of them were compatible with the serum profile of autoimmune hepatitis type-1. CONCLUSIONS: Liver involvement was complicated in 22.2% patients of primary Sjogren's syndrome. Clinical manifestations were non-specific and the main pathological change was chronic active hepatitis. The differential diagnosis between Sjogren's syndrome with liver involvement and type-1 autoimmune hepatitis could be only ascribed to other systemic clinical manifestations of Sjogren's syndrome. PMID- 10374421 TI - Clinicopathological study of renal involvement in patients with systemic sclerosis. AB - OBJECTIVE: To assess the incidence of renal involvement in patients with systemic sclerosis (SSc) as well as its clinical and pathological changes. METHODS: The renal involvement was studied clinicopathologically in 93 patients who were compatible with the diagnosis of SSc retrospectively. RESULTS: Eighteen patients (19.4%) were diagnosed as renal involvement by one or more of the following: proteinuria, renal hypertension, elevated levels of blood urea nitrogen (BUN) and/or serum creatinine (sCr). Renal impairment was observed in 5 patients (5.4%). The mortality rate was 12.9%, and 5 patients died of renal failure. Histological study was performed in 5 patients. The thickening of interlobular arterioles with intimal proliferation was found in 4 of the patients who also showed mild nonspecific glomerular changes. Two had no clinical features of renal involvement, 1 had renal hypertension and 1 died of renal failure. Another patient with a 22-year disease duration showed chronic glomerulonephritis with nephrosclerosis. CONCLUSIONS: SSc patients should be followed-up clinically and renal biopsy performed if necessary in order to discover early renal involvement and to insert rational therapy to improve its prognosis. PMID- 10374422 TI - The epidemiology study of hyperuricemia and gout in a community population of Huangpu District in Shanghai. AB - OBJECTIVE: To investigate the prevalence of hyperuricemia and gout in a community population of Huangpu District in Shanghai. METHODS: In the target community, 2037 dwellers were interviewed with relevan questionnares from house to house. According to even house number 1017 blood samples were taken for serum uric acid (SUA) levels measured with the uricase-peroxidase enzymatic method. RESULTS: The prevalence of hyperuricemia was 14.2% in men (SUA > 70 mg/L, 62 cases), 7.1% in women (SUA > 60 mg/L, 41 cases), 10.1% in both sexes. Seven gout patients were all men. The prevalence of gout in 2037 dwellers in Huangpu District was 0.77% in men and 0.34% in both sexes. CONCLUSIONS: The mean SUA level in each age group in this survey was much higher than that of a previous study 1 carried out in Shanghai, Beijing and Guangzhou in 1980 (P < 0.05). And the prevalence of hyperuricemia was increased rapidly (in men: from 1.4% in the survey of 1980 to 14.2% in our survey; in women: from 1.3% in the survey of 1980 to 7.1% in our survey). Compared with Idonesia data in 1992, the prevalence of hyperuricemia and gout in our survey was lower than that in Indonesia (P < 0.05), which suggests that racial and genetic predispositions are key causative factors. PMID- 10374423 TI - A better long-term outcome in cardiac transplant recipient with a history of previous open heart operations. AB - OBJECTIVE: To investigate the effect of previous open heart operations (POHO) on the outcome of heart transplantation (HTX). METHODS: Between November 1984 and May 1996, HTX was performed on 151 patients at Hartford Hospital. Among them, 61 patients had previous open heart operations (POHO) (group A), and 90 did not (group B). The average follow-up period was 1615 +/- 1185 days for group A and 1330 +/- 1125 days for group B. The recipient age was 55 +/- 10 years for group A and 48 +/- 12 years for group B (P < 0.01). There were 17 patients (26%) in group A and 14 (50%) in group B who were over 60 years of age. There was more coronary artery disease (74% versus 37%, P < 0.001) as etiology, and more diabetics in group A (P < 0.02). RESULTS: The time for cardiopulmonary bypass (133 +/- 20 min versus 106 +/- 18 min, P < 0.01) and aortic clamp time (73 +/- 16 min versus 61 +/- 13 min, P < 0.01) were longer in group A. The operative mortality (within 30 days) was 0 and 2.2%, and the cumulative deaths were 16 (26%) and 43 (48%) respectively for group A and group B (P < 0.01). The causes of death were (group A vs group B): infection (31% vs 26%), rejection (13% vs 28%, P < 0.05), malignancy (25% vs 16%), cardiac event (6% vs 14%) and others (25% vs 16%). In patients over 60, there were 4 deaths (24%) in group A and 7 (50%) in group B. The difference was not significant. No patients died of rejection in this subgroup. The actuarial survival rates in group A versus group B were: 1 year, 93% versus 83%; 2 years, 85% versus 74%; 3 years, 81% versus 71%; 5 years, 76% versus 58%; and 10 years, 57% versus 24% (P < 0.01). CONCLUSION: The survival rate in patients who had POHO is much higher than that in patients who had HTX as their primary operation. PMID- 10374424 TI - Analysis of the characteristics of folate binding proteins and its relationship with expression of multidrug resistance P-glycoprotein in myelodysplastic syndromes. AB - OBJECTIVE: To observe the characteristics of folate binding proteins (FBP) in myelodysplastic syndromes (MDS) and leukemia and to study the clinical significance of reduced folate carrier (RFC) present in MDS and its relationship with multidrug resistance (MDR). METHODS: The features of FBP on bone marrow cells were analyzed using radiolabeled 3H-folic acid (3H-FA) binding membrane proteins and SDS-polyacrylamide gel electrophoresis (SDS-PAGE). In the same time, P-glucoprotein and mRNA of MDR gene were detected using immunocytochemistry and reverse transcription polymerase chain reaction (RT-PCR) respectively in patients with MDS and leukemia. RESULTS: Two kinds of FBP, folate receptor (FR) and reduced folate carrier (RFC), were found on the leukemic cells. The same results were presented on mononuclear cells of bone marrow in 5 out of 14 MDS patients, and MDR positive was seen in 4 patiens of them. In normal control and other 9 cases of MDS FRs were only found on the mononuclear cells of bone marrow. CONCLUSION: Reduced folate carrier, which is present in the leukemic cell, is a product of neoplastic cell. It might reveal preleukmic state and have the same significance with MDR that RFC is found in MDS patients. PMID- 10374425 TI - Experimental study of lymph node auto-transplantation in rats. AB - OBJECTIVE: To observe the restoration of structure and function of auto transplanted lymph nodes. METHODS: Inguinal lymph nodes in Spregue-Dawley (SD) rats were auto-transplanted by free implantation, or with an intact vascular pedicle, or by free transplantation with microvascular anastomosis, to the popliteal fossa where lymph nodes were removed. The observation methods included emission computerized tomographic (ECT) scanning, staining of China ink and methylthionine chloride to observe the histological changes. RESULTS: After four weeks, these vascularized nodes showed normal histological appearances and spontaneously reestablished afferent and efferent lymphatic reconnection with the surrounding lymphatic vessels. ECT lymphoscintigraphy with 99mTc-Dx showed that vascularized lymph nodes had restored their normal function. CONCLUSION: Vascularized lymph node transplantation is a useful method for draining extremity lymph edema. PMID- 10374426 TI - Dynamic study of nitric oxide and endothelin-1 during endotoxin shock and effects of their antagonists on hemodynamics. AB - OBJECTIVE: To examine the relationship between the profound hypotension in endotoxic shock and the dynamic changes of nitric oxide (NO) and endothelin-1 (ET 1), so as to figure out which of the NO or ET-1 was more involved in the pathogenesis of endotoxic shock. And to investigate whether an offset of their opposite vasoactive effects would occur during endotoxic shock. METHODS: 24 rabbits were anesthetized and instrumented for recording hemodynamics. Endotoxin (E. coli 026: B6, 600 micrograms/kg) was bolus injected intravenously and the animals were randomly divided into four groups. Group I was control without any more intervention, and Group II, III, IV received bolus injections of L-NMA (10 mg/kg), phosphoramidon (2 mg/kg) or dexamethasone (2 mg/kg) respectively at 30 min post-endotoxin. Plasma NO3-, ET-1 and hemodynamics were measured at regular intervals. Their relationships were compared and analysed. RESULTS: Plasma ET-1 achieved its peak level at 60 min post-endotoxin, and then waned. Plasma NO3- started rising at 120 min post-endotoxin, then progressive increase continued till the last measurement at 180 min post-endotoxin. The decrease of blood pressure was significant at about 120 min post-endotoxin and further went down until death. The changes of hemodynamics and NO showed a quite close temporal correspondence between the increase of NO and the decrease of blood pressure. L NMA and phosphoramidon obviously reduced the plasma levels of NO and ET-1 to below their respective baseline levels, and showed transient effect of increase on blood pressure. Soon afterwards, however, the status of hemodynamics was aggravated. Dexamethasone just inhibited the excessive increase of NO and ET-1 during endotoxic shock without interfering their baseline levels and showed most beneficial effects on hemodynamics. CONCLUSIONS: Both NO and ET-1 increase during endotoxic shock, but only the increase of NO has a close temporal correspondence with the decrease of blood pressure. It suggests a more important role of NO in pathogenesis of endotoxic shock. The increase of NO and ET-1 is different in time process, which indicates that an offset of their opposite vasoactive effects would not occur. Intreference against the increase of NO and ET-1 during endotoxic shock is most beneficial when their baseline levels are maintained. PMID- 10374427 TI - Effectiveness of the analogue of natural Schisandrin C (HpPro) in treatment of liver diseases: an experience in Indonesian patients. AB - OBJECTIVE: To determine the effect of dimethyl-4,4'-dimethoxy-5,6, 5',6 dimethylene dioxybiphenyl-2,2'-dicarboxylate (HpPro) on patients with acute and chronic liver diseases. METHODS: An open trial and a prospective randomized and controlled study were performed. The open trial consisted of 56 cases (16 cases of acute hepatitis, 20 cases of chronic hepatitis, 14 cases of liver cirrhosis and 6 cases of fatty liver). Controlled study consisted of 20 cases of Child A chronic hepatitis which were randomly treated with either HpPro or a mixture of known drugs which used as a liver protective agent in Indonesia as control for one week. The patients were then crossed over those two drugs in the next week. RESULTS: In the open trial, after 4 weeks' treatment with HpPro 7.5 mg orally three times daily, acute hepatitis, chronic hepatitis and fatty liver cases showed rapid decrease of SGOT and SGPT. In the liver cirrhosis cases, SGOT and SGPT were decreased slowly. In the controlled trial, nine patients received HpPro 7.5 mg three times daily orally and eleven were treated with a mixture of known drugs as the controls. After one week treatment, HpPro group clinically showed significant decrease of SGPT and SGOT levels compared to control group (P = 0.035). At the second week, HpPro group showed significant decrease of SGOT compared to control group (P = 0.038) but the decrease of SGPT was not significant (P = 0.096). CONCLUSION: Treatment with HpPro is effective to reduce liver impairment in acute and chronic liver diseases on Indonesian patients. No side effect of HpPro was observed. PMID- 10374428 TI - Immunohistochemical studies on the endotoxin-induced uveitis. AB - OBJECTIVE: To investigate the longitudinal changes of macrophages and major histocompatibility complex (MHC) class II-positive cells in the iris and ciliary body of Lewis rats after lipopolysaccharide (LPS) injection. METHODS: Immunohistochemistry was performed using monoclonal antibodies to monocytes and macrophages (ED1) and MHC class II-positive cells (OX6) on wholemounts of the iris and ciliary body in endotoxin-induced uveitis (EIU). RESULTS: A network of macrophages (ED1+ cell) and MHC class II-positive cells, was present in the iris and ciliary body of normal Lewis rats. Most cells in the iris and ciliary body displayed dendritiform appearance. A severe involvement of the iris and ciliary body, as evidenced by a rapid influx of monocytes and macrophages and remarkable increase of MHC class II-positive cells, was observed after LPS injection. CONCLUSIONS: A network of macrophages and MHC class II-positive cells in the iris and ciliary body may play an important role in immune surveillance. LPS injection induces a severe inflammation in the anterior segment of the eye, which may serve as a model for acute anterior uveitis in human. PMID- 10374429 TI - Obstructive sleep apnea syndrome: an experience in Chinese adults in Hong Kong. AB - OBJECTIVE: Epidemiologic studies in Caucasian populations suggested that symptomatic obstructive sleep apnea (OSA) occurred at a prevalence of 1-10%. The condition has been increasingly recognised among the Chinese in Hong Kong. We therefore, summarize our experience with OSA at the Department of Medicine, The University of Hong Kong at Queen Mary Hospital from 1985-1996. METHODS: All clinic records concerning demographic data, anthropometric data, clinical features, polysomnographic findings and treatment were reviewed. RESULTS: One hundred and twenty-two patients were diagnosed to have OSA. Demographic and clinical features were similar to Caucasian data with a male predominance of 84%, a mean age of about 50 years, and obesity as a risk factor at a mean body mass index of 30.4, which was higher than that of the average local population, although lower than that of OSA patients in Caucasian series. About 27% of the patients have a body mass index (BMI) similar to or below the population average. Nearly all were habitual snorers, and the majority had excessive daytime sleepiness. On polysomnography, the mean apnea-hypopnea index was 38. Common associated medical conditions were hypertension (34%), diabetes mellitus (10%), ischemic heart disease (9%), hyperlipidemia (6%). Most patients were managed successfully with nasal continuous positive airway pressure. Treatment with oral appliances for milder cases is being explored. CONCLUSIONS: OSA has been increasingly recognised among Chinese adults in Hong Kong in the past decade. Demographic features were similar to Caucasian data. The majority of patients were overweight, although 27% were not, and further investigation on the contribution of faciomaxillary morphology to OSA in this group is warranted. PMID- 10374430 TI - Study on the transmission threshold value of bancroftian filariasis. AB - OBJECTIVE: To elucidate the transmission dynamic and epidemic trend of bancroftian filariasis occurred under the condition with no control measure taken 5 years after elimination of filariasis. METHODS: A 10-year longitudinal observation (from 1984 to 1994) was made in Huayuan Village in Shengli Township of Tancheng County, which used to be a high bancroftian filariasis-endemic area in southern part of Shandong Province. RESULTS: The microfilarial rate decreased from 0.56% before the study to 0.12% after the study and 8 out of the 9 previous microfilaria-positive cases became negative gradually. During the study period, 6 new microfilaremia cases were detected, 5 of which became negative naturally within 3 to 4 years. Eighty-eight point eight nine per cent of the detected patients with microfilaremia converted into IgG4-negative after 10 years. The natural infective rate of vectors decreased year by year and became zero by the tenth year of the study, the annual transmission potency decreased also from 3.47 to zero by the tenth year. CONCLUSIONS: It showed that under the local natural environment the biting rate representing the vector density which was obtained by capture method was from 24.1 to 52.5 person/night among the residents who did not use mosquito nets, and 13.5 to 21 person/night among the residents who used mosquito nets. The microfilarial rate of 0.56% in population with the average microfilarial density of 6.6 to 20.7 capita/60 microliters ear blood of residual microfilaria-positive patients might be considered as the terminal threshold of transmission. PMID- 10374431 TI - Effect of interleukin-6 on the growth of human lung cancer cell line. AB - OBJECTIVE: To investigate the effect of interleukin-6 (IL-6) on the growth of human lung cancer in vivo as well as in vitro. METHODS: To examine the mRNA level of IL-6 receptor (IL-6R) in high-metastatic human lung giant cell carcinoma cell line PG by means of reverse transcription polymerase chain reaction (RT-PCR). To assess the existence of IL-6 receptor complex (including IL-6R and gp130) with the treatment of PG cells by use of recombinant human IL-6 (rhIL-6), recombinant human oncostatin M (rhOSM), and recombinant human leukemia inhibitory factor (rhLIF), respectively. To detect the expression of IL-6 by Northern blotting hybridization and bioactive assay. To identify the effect of IL-6 secreted by PG cells by use of IL-6 and IL-6R antisense oligodeoxynucleotides (ODNs), and specific neutralizing antibody to IL-6. To document the influence of IL-6 on PG cells growth in vivo through the strategy of the transfection of expression vector inserted antisense IL-6 cDNA. RESULTS: RT-PCR analysis revealed that PG cells expressed IL-6R mRNA. Any one of the recombinant cytokine IL-6, OSM and LIF stimulated the growth of PG cells in vitro in a concentration-dependent manner. These results demonstrated IL-6 receptor complex exist in PG cells. At the same time, PG cells expressed IL-6 mRNA and secreted bioactive IL-6. Both IL-6 antisense ODNs and IL-6R ODNs inhibited PG cells proliferation. Treatment of PG cells with IL-6 antibodies reduced the growth of PG cells in vitro. PG cells transfected with IL-6 antisense expression vector showed a decreased growth in nude mice. CONCLUSION: IL-6 functions as an autocrine growth stimulator for PG cells in vivo as well as in vitro. PMID- 10374433 TI - Evaluation of second cytoreductive surgery in the treatment of epithelial ovarian cancer. AB - OBJECTIVE: To evaluate the effectiveness of second cytoreductive surgery in the treatment of epithelial ovarian cancer. METHODS: From January 1989 to June 1994, second cytoreductive surgery was carried out on 33 patients with epithelial ovarian cancer who either underwent unsatisfactory primary debulking operation or had recurrence. According to FIGO staging (1987), there were 5 patients in stage I, 2 in stage II, 25 in stage III, and 1 in stage IV. Pathological grading was G1 in 2 cases, G2 in 9, G3 in 19 and uncertain in 3. The 33 patients can be divided into 3 categories: I, nine patients who had unsatisfactory primary debulking operation with macroscopic residual > 2 cm, and 1-2 courses of postoperative chemotherapy; II, 15 patients who had 6-8 courses of cisplatin-based postoperative chemotherapy and in whom recurrence was diagnosed after complete response for at least 3 months; and III, 9 patients who had the same treatment as category II and survived without cancer clinically for more than 6 months, and in whom recurrence was diagnosed during second-look laparotomy. All patients had been followed up for at least two years (27-168 months) dated from the primary debulking operation. RESULTS: Fifteen cases had no macroscopic residuals (group A), 5 had residuals < 2 cm (group B), and 13 had residuals > 2 cm (group C). The medium survival time and two-year survival rate in groups A, B and C were 59.09, 20.6 and 8.29 months, and 93.3%, 20% and 7.69% respectively (P < 0.001, A vs C; P < 0.05, A vs B and B vs C). CONCLUSIONS: The results suggest that second cytoreductive surgery is of value, and the key to success is to eliminate any macroscopic residual focus, or at most, to leave only minimal residuals < 2 cm. It is suggested that well-targeted multiple-route chemotherapy with sufficient courses before second cytoreductive surgery is important to achieving better results. PMID- 10374432 TI - CD 80(B7-1) expression on human tumor cell lines and its costimulatory signals for T cell proliferation and cytokine production. AB - OBJECTIVE: To investigate CD 80 expression on human tumor cell lines and establish stable CD 80 expression transfectants to illustrate CD 80 costimulation on the T cell proliferation and cytokine production. METHODS: Raji, MDA-453, MCF 7, Hela, 3AO, MKN-45 and EBV transformed B cell were detected for CD 80 expression by RT-PCR. CD 80 cDNA subcloned to retrovirus vector pLXSN, with them stable CD 80 transfectanants were established. With specific mAb BB1 and FACS assay, the expression of CD 80 were detected. Anti-CD3 induced T lymphocyte proliferation and IL-2, IFN-r production were performed to evaluate CD 80 costimulatory activity in the presence of calcium ionophore A23187 and phorbol ester PMA. RESULTS: CD 80 (B7-1) expression of MDA-453, MCF-7, Hela, 3AO, MKN-45 was negative, and that of Raji and EBV transformed B cell was positive by RT-PCR and FACS methods. Cocultured with CD 80 transfected human breast carcinoma cell line MDA-453, anti-CD3 induced T cells proliferation and cytokine production were significantly increased in the presence of A23187 and PMA, but not with parental MDA-453. CONCLUSIONS: CD 80 expression is absent on most human tumor cell line except for Raji and EBV transformed B cell. Full activation of T cell needs CD 80 costimulation. Influence of CD 80 costimulation on cytokine production is mainly regulated via IL-2 and IFN-r. PMID- 10374434 TI - The test of motion perception of the normal Chinese subjects. AB - OBJECTIVE: To investigate the characteristic of motion perception (MP) of normal Chinese subjects. METHODS: MPs were recorded from 56 normal subjects (112 eyes), age ranged from 11 to 68 years, and the MP software was controlled by PC compatible computer which appeared as the vertical bar targets in VGA screen. RESULTS: The Mp rates were ascending gradually from 10 age-group to 30 age-group as the age increased in the 2 pixels (2P) horizontal motion, and the MP rates were decreasing gradually over 40 age-group, and the MP rates were not affected by the age in the 4 pixels (4P), 6 pixels (6P) horizontal motion and > 40 Hz flick motion. There was no relation between the MP and the sex and the different eyes. CONCLUSIONS: The characteristic of MP of the normal Chinese subjects was determined and the results provided the normal reference values. PMID- 10374435 TI - Incidence and predictive value of congenital hypertrophy of retinal pigment epithelium in Chinese familial adenomatous polyposis patients. AB - OBJECTIVE: To investigate the incidence and predictive value of congenital hypertrophy of retinal pigment epithelium (CHRPE) in the Chinese familial adenomatous polyposis (FAP) patients. METHODS: Eleven FAP patients and 28 at risk relatives from 7 families were examined for CHRPE lesions. RESULTS: CHRPE was present in all 11 FAP patients. Nine CHRPE lesions were found in 9 of the 28 at risk relatives. There were great intra and inter-familial variations of the CHRPE lesions. Two at risk relatives were diagnosed to have FAP only after our screening and subsequent proctosigmoidoscopy. Nine at risk relatives were put under high surveillance because of the presence of CHRPE. CONCLUSION: The incidence of CHRPE in the FAP patients in our study is 100%. Eye examination for CHRPE for FAP patients and their at risk relatives is of very high value. A central FAP registry is thus recommended. PMID- 10374436 TI - Research on nucleolar organizer regions of hippocampal neuron in Alzheimer's disease. AB - OBJECTIVE: To understand the cellular genetic expression of the cell's population by studying the nucleolar organizer regions (NORs) of hippocampal neuron of Alzheimer's disease (AD). METHODS: The postmortem human hippocampal tissues were divided into three groups, namely, the young, the elderly and the AD groups. Each group contained tissues from 10 patients. The study was conducted using image pattern analysis of the nucleoli-nucleoplasms ratio of the neurons of Nissl's stained pathological cerebral hippocampal tissues, the area of stain, and the integrating absorption of nucleoli of silver-stained NORs. RESULTS: The nucleoli nucleoplasms ratio of the neurons of Nissl's stained cerebral hippocampal tissues, the area of stain, and the integrating absorption of the nucleoli of hippocampal neuron were decreased in the elderly and the AD groups as compared with the young group. However, the area of stain and the integrating absorption of the nucleoli of the hippocampal neurons were relatively increased in the AD group in comparison with the elderly group. CONCLUSION: Nissl's stain demonstrates the hypofunction of the hippocampal neurons in the elderly and the AD patients. The Silver stain of NORs shows the decline of rDNA transcription activity of the nucleoli of the hippocampal neurons in the elderly and the AD patients. However, the transcription activity of the nucleoli of the hippocampal neurons of AD patients was relatively improved, and the cellular genetic expression of the cell's population was relatively strengthened. These cellular morphological changes have probably reflected the cellular defensive system. PMID- 10374437 TI - Injury of liver sinusoidal endothelial cells in isolated liver perfusion model for regional chemotherapy in rats. PMID- 10374438 TI - Anastomosis of duodenal mucosa with gastric wall: a report of 45 cases. PMID- 10374439 TI - [Improvements in the treatment and management of control the occurrence of multidrug-resistance tuberculosis]. PMID- 10374441 TI - [Construction of Escherichia coli-Mycobacteria shuttle plasmid and the stable expression of human interleukin-2 in BCG and Escherichia coli]. AB - OBJECTIVE: To construct and identify Escherchia coli (E. coli)-Mycobacteria shuttle plasmid and to detect stable expression of foreign gene in E. coli and BCG. METHOD: With a genetic engineering technique to construct the E. coli Mycobacteria shuttle plasmid, the human interleukin-2 (IL-2) gene was electrophoreted into BCG with recombinant plasmid PZSIII-I and positive clones selected using polymerase chain reaction (PCR) technique. The expression of foreign gene of human IL-2 in BCG was identified by ELISA and SDS-PAGE. RESULT: Human IL-2 cytokine was steadily expressed in recombinant BCG and E. coli and could secrete outside the cell. CONCLUSION: M. bovis BCG recombinant constructed can produce and secrete the human IL-2. A secretion of the active cytokine was accomplished through the combined use of the BCG HSP65 promoter and a secretion signal from the BCG Ag-85B. The BCG HSP65 promoter is active in both BCG as well as E. coli which can not secrete foreign protein. PMID- 10374440 TI - [Study on the molecular mechanism of multi-drug resistance in clinical isolates of Mycobacterium tuberculosis]. AB - OBJECTIVE: To study the molecular mechanism of multi-drug resistance in M. Tuberculosis, and to develop a new method for detecting genes related with multi drug resistance. METHOD: The ropB, rpsL, katG genes and inhA regulatory sequence in clinical isolates of M. tuberculosis were analyzed with PCR and PCR-SSCP techniques. RESULT: The sensitivity of amplifing the drug-resistant genes with PCR was 1-10 pg DNA. Of the 20 multiple resistant strains with reduced sensitivity to streptomycin, rifampin and isoniazid, 90% showed mutations in more than two genetic markers associated with resistance to each of these three drugs, 10% revealed only mutations in rpoB gene. CONCLUSION: Multi-drug resistance in M. tuberculosis could be caused by an accumulation of mutations in chromosomal genes encoding drug targets or an alteration at a single multiple resistance locus. PCR and PCR-SSCP techniques might become simple, rapid and reliable diagnostic tests for multi-drug resistance. PMID- 10374442 TI - [The correlation between lung cancer lymph node metastasis and nm23-H1 gene mutation, mRNA expression]. AB - OBJECTIVE: To examine the genomic status and the mRNA expression of the nm23-H1 gene, and to analyse the relationship between metastasis of non-small cell lung carcinomas and the gene abnormalities. METHOD: By using PCR-SSCP and semiquantitative RT-PCR, the nm23-H1 gene mutation and mRNA expression were studied in 31 cases of non-small cell lung cancer. The normal tissue adjacent to carcinoma and normal lung tissue were used as control. RESULT: None of the nm23 H1 gene mutation was found in all the lung cancer samples. Decreased expressions of this gene were found in 14 of 20 lung cancer cases with lymph node metastasis, the rate (14/20) of this was significantly higher than that (3/11) of the non metastatic cancers (P < 0.05). CONCLUSION: These data indicate that nm23-H1 may be a putative metastasis suppressor gene which had shown reverse regulating activation in the metastatic progression of lung cancers, and the level of nm23 H1 mRNA may be considered as one of the pathological indicators in predicting metastatic potential of lung cancers. PMID- 10374443 TI - [Expression and distribution of bFGF in rat lung tissue of chronic hypoxic pulmonary hypertension]. AB - OBJECTIVE: To evaluate the role of bFGF in the development of hypoxic pulmonary hypertension. METHOD: Rat models with chronic hypoxia induced pulmonary hypertension were established, the pulmonary hemodynamics were measured and the pulmonary arterioles change were studied with morphometric analysis under light microscopes, immunohistochemical staining with monoclonal antibody against human recombinant bFGF was performed in the paraffin section of rat lung. RESULT: (1) The mean pulmonary artery pressure (mPAP), and the ratio of the thickness of pulmonary arteriolar wall to external diameter of pulmonary arterioles (MT%) were 3.96 +/- 0.47 kPa and 33.8% +/- 3.5% in rats exposed to hypoxia for 3 weeks respectively, both were significant higher than those in normal control group, P < 0.01. (2) The positive staining for bFGF in the wall of pulmonary arterioles in hypoxic rats was stronger than that of control group (P < 0.01), there was a statistical relationship between increase of staining for bFGF and MT% in rats exposed to hypoxia. CONCLUSION: (1) Hypoxia can induce formation of pulmonary hypertension and structual remodeling of pulmonary arterioles. (2) bFGF may modulate the structure remodeling of pulmonary arterioles in chronic hypoxic pulmonary hypertension. PMID- 10374444 TI - [The therapeutic effect of TGF-beta monoclonal antibody to bleomycin-induced pulmonary fibrosis in rats]. AB - OBJECTIVE: Transforming growth factor-beta (TGF-beta) plays an important role in the pathogenesis of interstitial lung fibrosis. It can stimulate indirectly the mitosis of lung fibroblasts (LFb) as well as the synthesis and disposition of extracellular matrix. The purpose of this study was to evaluate the effects of TGF-beta monoclonal antibody on bleomycin-induced interstitial pulmonary fibrosis. METHOD: The effect of TGF-beta monoclonal antibody to bleomycin-induced pulmonary fibrosis in rat was studied either in vivo or in vitro. The proliferation of lung fibroblasts was studied by measuring the incorporation rate of 3H-TdR. Northern hybridization was used to detect mRNA level of procollagen A1 (I) and procollagen A1 (II) in cultured LFbs and lung tissue. RESULT: The results showed that the incubation supernatant of alveolar macrophages from rats with pulmonary fibrosis had the ability to stimulate fibroblast proliferation as well as their procollagen expression. The monoclonal antibody could inhibit the proliferation of LFbs in a concentration-dependent manner. The highest concentration, 100 micrograms/ml, of TGF-beta antibody could inhibit incorporation rate from 1749 +/- 322 of the fibrosis group to only 833 +/- 277 (P < 0.01). The mRNA level of procollagen I and II was decreased by 44% and 28% respectively after the antibody treatment. In vivo, procollagen I and II mRNA level were decreased by 40% and 12% after the administration of TGF-beta antibody though it could only slightly alleviate the extent of fibrosis and alveolitis in lung tissue. CONCLUSION: TGF-beta is a key factor stimulating the proliferation and collagen synthesis. TGF-beta antibody can partially neutralize its action. This may suggest that the antibody might become a new therapeutic choice for pulmonary fibrosis in the future. PMID- 10374445 TI - [Experimental study on prevention of pneumoconiosis complicating with tuberculosis by Mycobacterium vaccae]. AB - OBJECTIVE: To explore the role of Mycobacterium vaccae in prevention of pneumoconiosis complicating with tuberculosis. METHOD: Mycobacterium vaccae was injected into rats which had been exposed to quartz for 2 weeks, and H37Rv was injected into their tail veins 1 month later. All the rats were killed 3 months later. Indexes for evaluation in the study included tuberculous lesion pathological change index, Mycobacterium tuberculosis culture in a fixed amount in lung tissues, count of Mycobacterium tuberculosis and histopathological changes in alveolar macrophages. RESULT: Tuberculous lesion pathological change index, Mycobacterium tuberculosis culture in a fixed amount in lung tissues and count of Mycobacterium tuberculosis in alveolar macrophages were 2.6 +/- 0.5, (4.40 +/- 4.00) x 10(4) CFU and 7.2 +/- 3.2 respectively in pneumoconiosis complicated with tuberculosis and injecting Mycobacterium vaccae group, while 3.1 +/- 0.3, (18.9 +/- 18.2) x 10(4) CFU and 12.5 +/- 6.3 respectively in the control group. And statistically significant differences were found between the two groups (P < 0.05, 0.01, 0.05 respectively). The histopathological analysis revealed that proliferative and lymphoid nodules were predominant in the pneumoconiosis complicated with tuberculosis and injecting Mycobacterium vaccae group, while necrotic nodules in the control group. CONCLUSION: Mycobacterium vaccae might play a role in prevention of pneumoconiosis complicating with tuberculosis. PMID- 10374446 TI - [Bronchofiberscope and catheter intervention in treatment of multi-drug resistant pulmonary tuberculosis]. AB - OBJECTIVE: To evaluate the clinical value of bronchofiberscope and catheter intervention in treatment of multi-drug resistant pulmonary tuberculosis. METHOD: Forty-eight patients with multi-drug resistant pulmonary tuberculosis were treated by injecting ofloxacin and amikacin through bronchofiberscope and catheter in addition to chemotherapy, while forty controls were treated by chemotherapy only. RESULT: At the end of the treatment, the sputum conversion rate was 92%, radiographic improvement rate 96% and cavity closing rate 27% in the treatment group, all of which were higher than the controls (63%, 58% and 10% respectively) (P < 0.01-0.05). No complication and obvious adverse reaction were observed. CONCLUSION: The efficacy of bronchofiberscope and catheter intervention in addition to chemotherapy is better than only chemotherapy in treatment of multi-drug resistant pulmonary tuberculosis. PMID- 10374447 TI - [Efficacy of unfixed continuation phase short-course chemotherapy]. AB - OBJECTIVE: To evaluate the efficacy of the short-course chemotherapy shorter than six months. METHOD: The 2SHRZ/xHR regimen was used in 290 smear positive new untreated tuberculosis patients. The duration of continuation phase was not fixed, just in accordance with the speed of sputum negative conversion. The treatment was continued until sputum negative conversion mantained for three consecutive months. RESULT: Two hundred eighty-three out of 290 patients were cured. The sputum negative conversion rate at the end of the sixth month was 98.3%. The duration of treatment was 4.7 months on the average. The rate of two year follow-up was 94.3%, and the rate of bacteriological relapse during two-year follow-up was 1.9%. CONCLUSION: The results showed that the length of the six month short-course chemotherapy could be shortened, and its recent and long-term efficacy was found satisfactory. PMID- 10374448 TI - [Analysis of 9 cases of pulmonary tuberculosis complicated with legionnaires disease]. AB - OBJECTIVE: To study the characteristics of pulmonary tuberculosis complicated with legionnaires disease (LD) to avoid misdiagnosis and incorrect treatment. METHOD: Nine cases of pulmonary tuberculosis complicated with LD were retrospectively analyzed. RESULT: The clinical and chest X-ray manifestations varied, and no characteristics were found in these cases. Because cross antibodies existed between Legionella pneumophila and other causal bacteria, it was found difficult to differentiate LD, pulmonary tuberculosis and other causal bacteria infection. Efficacy of erythromycin combined with rifampicin, and decrease of serum titres of Legionella pneumophila four times after treatment were found helpful for definite diagnosis of LD. CONCLUSION: Only paying much attention to LD, and detecting the serum antibody as early as possible can provide evidence for diagnosing of the disease. PMID- 10374449 TI - [Advances in the study on the mechanism of pleural effusion]. PMID- 10374450 TI - [The results of questionnaire of basic concepts about asthma management and prevention for respiratory professionals in Beijing's hospitals at different levels]. AB - OBJECTIVE: To evaluate the information of basic concepts about asthma management and prevention collected from respiralogy professionals in Beijing's hospitals at different levels and provide the basis of drawing up a work plan for Asthma Management and Prevention Committee. METHODS: Fifty-five hospitals including municipal hospitals, urban district hospitals and county hospitals of suburban district were involved and four hundred and thirty-six respiralogy professionals consisting of resident physician and physician-in-charge were asked using questionnaire method. RESULTS: The respiralogy professionals at different level hospitals have some confused ideas in the definition of asthma, the importance of inhalation and anti-inflammatory therapy and the value of peak flow meter. The results of quetionnaire of respiralogy professionals in municipal hospitals were superior to that in urban district hospitals and county hospitals of suburban district. CONCLUSION: The physician education of asthma management and prevention is an important task. Special attention should be paid on the hospitals at basic level. PMID- 10374451 TI - [The effect of GM-CSF on endothelin-1 and endothelin converting enzyme gene expression in human airway smooth muscle cells with or without theophylline incubation]. AB - OBJECTIVE: To explore the role of airway smooth muscle cells (ASMC) stimulated by inflammatory factors in the development of asthma and bronchial hyperresponsiveness (BHR) and the possible mechanism of low dose theophylline in the treatment of airway inflammation, the effects of GM-CSF on endothelin-1 and endothelin converting enzyme (ECE) gene expression in human ASMC and theophylline on their expressions were investigated. METHOD: The cultured human ASMCs were treated with GM-CSF (1,000 micrograms/ml), GM-CSF plus theophylline (125 ng/ml) respectively for 8 hours. Endothelin-1 and ECE gene expresion were measured by RT PCR. RESULT: There was no ET-1 gene expression in control group, but strong expression in GM-CSF treated group; theophylline entirely inhibited GM-CSF induced ET-1 gene expression in the ASMC; no significant difference of ECE gene expression was found in three groups. CONCLUSION: GM-CSF causes a strong ET-1 gene expression but not ECE gene expression in cultured human airway smooth muscle cells; theophyllin inhibites this abnormal expression which may be one of the anti-inflammatory mechanisms of low dose theophylline in the management of chronic airway inflammation. PMID- 10374452 TI - [The relationship between interleukin-8 and airway hyperresponsiveness]. AB - OBJECTIVE: To explore the relationship between interleukin-8 (IL-8) and airway hyperresponsiveness (AHR) of asthma. METHOD: IL-8 at a dose of 0.5 microgram/kg or 5 micrograms/kg was administered intranasally to guinea-pigs twice a week for 3 weeks. 24 hours after the last administration, airway responsiveness was measured as an overall index of airway response to increasing concentrations of inhaled histamine (25, 50, 100 and 200 micrograms/ml) and the numbers of different inflammatory cells in bronchoalveolar lavage fluid (BALF) was counted. RESULTS: The IL-8 treatment significantly enhanced airway responsiveness to histamine in a dose-dependent manner (P < 0.05 or 0.01) and induced a significant influx of neutrophils in BALF (P < 0.01), and there are many neutrophils within airway wall but not control animals treated with phosphate buffered saline (PBS). CONCLUSION: The IL-8 given into the airways can produce neutrophil inflammation of the airways and induce AHR, it may play an important role in asthma, especially in the development of AHR. PMID- 10374453 TI - [Chronic airway inflammation and atopic features in cough variant asthma]. AB - OBJECTIVE: To elucidate the chronic airway inflammation and atopic features in 17 patients with cough variant asthma (CVA). METHODS: (1) Allergen skin prick test, and the atopy index (AI) calculation. (2) Fiberoptic bronchoscopy, inflammatory score of airway membrane under bronchoscopy. (3) Biopsy of the airway membrane and calculation of the eosinophil infiltration. (4) Bronchoalveolar lavage, classification of various cells in the BALF. (5) House dust mite and rabbit antihuman IgE induced histamine release from mast cell. The results were compared with those in 9 patients with typical asthma (Group 8) and 7 normal subjects (Group C). RESULTS: The number of EOS and mast cells in BALF and the AI of CVA group were found higher than in control group. CONCLUSION: (1) Similar to typical asthma, IgE dependent type I allergic reaction plays an important role in the pathogenesis of CVA. (2) Chronic inflammation including eosinophil infiltration was shown in airway membrane in CVA. (3) Bronchial hyperresponsiveness and chronic inflammation of the airway membrane in CVA are less severe than those in typical asthma. (4) In patients who are in difficulty of making a diagnose of CVA clinically. BAL and biopsy of the bronchial membrane will help to make a definite diagnosis. PMID- 10374454 TI - [Aerosolized recombinant interferon-gamma prevent antigen-induced eosinophil recruitment in guinea pig trachea]. AB - OBJECTIVE: In order to determine whether interferon-gamma (IFN-gamma) inhibits eosinphil infiltration in the trachea of asthmatic guinea pigs induced by Rhizopus nigricans. METHOD: We had administered aerosolized rIFN-gamma in the tracheas of 30 sensitized guinea pigs which had been divided into six groups, then teated animal inhaled rIFN-gamma of 5 x 10(4), 20 x 10(4), and 40 x 10(4) concentration, BDP and normal saline respectively at 24 h, 12 h, 2 h before being challenged. RESULT: (1) Provocation positive rates decreased in 40 x 10(4) rIFN gamma and BDP group compared with that in normal saline group and before intervention (P < 0.05), airway resistence decreased (P < 0.01). (2) The administration of aerosolized rIFN-gamma (40 x 10(4)) and BDP also decreased fungus-induced eosnophils but not other cells infiltration in the trachea. (3) In BALF, Eos count and ECP level were obviously lower than those in other groups. However, eosinophil numbers did not show significant change in the peripheral blood. CONCLUSION: Local administration of rIFN-gamma (40 x 10(4)) may reduce airway inflammation and intervene asthmatic attack by inhibition of Eos, ECP infiltration in airways. PMID- 10374455 TI - [mRNA expression of granulocyte-macrophage colony-stimulating factor in airway tissues of asthma guinea pigs: effect of triptolide]. AB - OBJECTIVE: To explore the role of granulocyte-macrophage colony-stimulating factor (GM-CSF) in eosinophil inflammation of asthma airway. METHODS: Guinea pig models of asthma were established with aerosolized ovalbumin. A group of asthmatic guinea pigs were treated with injection of triptolide, density of eosinophil infiltration in airway tissues was observed under microscope and expression of GM-CSF mRNA in airway tissues was detected with dot hybridization by Dig-labeled cDNA probe. RESULTS: Expression of GM-CSF mRNA in asthmatic animals were higher markedly than that in the triptolide-treated and the control groups. However, there was no statistically differences of density of eosinophils infiltration between the asthma and the triptolide-treated groups. CONCLUSIONS: GM-CSF might participate in eosinophil inflammation in airway of asthmatic animals and triptolide might be of potential value in anti-inflammation for asthma. PMID- 10374457 TI - [The effect of protheo on pulmonary function and the change of its plasma concentration in patients with chronic asthmatic bronchitis]. AB - OBJECTIVE: In order to study the effect of protheo on pulmonary function and the change of its plasma concentration in patients with chronic asthmatic bronchitis. METHOD: 31 patients with chronic asthmatic bronchitis were treated by taking protheo 400 mg once a day orally for 6 weeks. The pulmonary functions, including FEV1, FVC, FEV1/FVC and PEFR, and the plasma concentrations of theophylline were measured respectively before and after the administration. RESULTS: The results showed that the pulmonary functions after the therapy were greatly improved compared with that before the therapy, even after the administration for two weeks. In the second, fourth, sixth week of the therapy, the plasma levels of theophylline after taking protheo for 4 hours and 12 hours were both within the effective concentration. CONCLUSION: Protheo is a kind of medicine with good effect, steady plasma concentration and less side effect for the patients with chronic asthmatic bronchitis. PMID- 10374456 TI - [Effects of theophylline on lung function and sleep in patients with severe chronic obstructive pulmonary disease]. AB - OBJECTIVE: To evaluate the effects of theophylline given by twice-daily and once daily on 24 hour lung function and sleep in severe COPD. METHOD: Ten patients were assessed. Before theophylline given, the baseline measurements on lung function and polysomnography were performed, and baseline scores of symptoms and sleep quality were investigated. And then, with double-blind crossover in design, twice-daily theophylline doseing (TD) or once-daily theophylline dosing (OD) was given in a randomly assigned sequence, and serum theophylline concentration (STC) was measured, and other physiological testings were similar as baseline measurements. RESULT: The peak STC occured at 4 am after administration at 8 pm, and that occured at 12 am after administration at 8 am, and the trough STC occured at 8 pm in both treatment. STC of OD was higher than that of TD at 4 am, but TD was higher than OD at 4 pm (P < 0.05). There was a similar but significant improvement of the lung function after theophylline given for TD or OD (P = 0.0001). Symptoms improved in both groups, but OD was better (P = 0.0023). The sleep stages and quality variation were not significant. CONCLUSION: There was higher STC in early morning in both treatment. The improvement of lung function may well be obtained at lower STC. There were not significant effect on sleep. PMID- 10374458 TI - [Evaluation of different methods of detection and diagnosis for infectious pulmonary tuberculosis]. AB - OBJECTIVE: To evaluate the compare the efficiency and benefit of fluoroscopy and direct sputum examination. METHOD: The suspected persons with pulmonary tuberculosis symptoms were enrolled to be examined with fluoroscopy, chest radiography, sputum smear and culture. RESULT: The diagnostic procedure used by World Bank-loaned Tuberculosis Control Project in China is based on fluoroscopy screening. The miss-detection rates of smear positive, culture positive and bacteriological positive pulmonary tuberculosis were 10.5%, 28.3% and 28.2% respectively. Its accuracy of diagnosis was lower than that of direct sputum smear examination and the cost was higher. CONCLUSION: Direct sputum smear examination seems to be the best diagnostic method for infectious pulmonary tuberculosis and suitable for application in rural areas. PMID- 10374459 TI - [Evaluating clinical significance of tumor necrosis factor-alpha and hydroxyproline in blood of patients with active pulmonary tuberculosis]. AB - OBJECTIVE: To evaluate the clinical significance of tumor necrosis factor-alpha (TNF-alpha) and hydroxyproline (HYP) in blood of patients with active pulmonary tuberculosis (APT). METHOD: Contents of TNF-alpha and HYP in blood of 28 patients with APT and 17 normal subjects were determined with enzyme-linked immunosorbent assay and ultraviolet spectrophotometer respectively. RESULT: It was found that the concentrations of TNF-alpha and HYP in patients with APT before antituberculosis treatment were obviously higher than those of the normal subjects (192.85 +/- 37.14 ng/L vs 89.36 +/- 23.18 ng/L and 4.96 +/- 1.13 mg/L vs 1.64 +/- 0.33 mg/L respectively, all P < 0.001) and those who received 3- month anti-tuberculosis treatment (192.65 +/- 37.14 ng/L vs 112.50 +/- 44.93 ng/L and 4.96 +/- 1.13 mg/L vs 2.17 +/- 0.39 mg/L respectively, all P < 0.001). TNF-alpha level was significantly correlated with HYP (r = 0.5132, P < 0.001). CONCLUSION: The results indicate that TNF-alpha might play an important role in inducing pulmonary tissue damage of APT, and the lowering of HYP content in blood might indicate that pulmonary tuberculosis is recovering from active stage. PMID- 10374460 TI - [Application of membrane lipid fluidity and superoxide dismutase in differential diagnosis of pulmonary tuberculosis and lung cancer]. AB - OBJECTIVE: To evaluate the application of membrane lipid fluidity and superoxide dismutase (SOD) in differential diagnosis of pulmonary tuberculosis and lung cancer. METHOD: Fluorescence-labeling (DPH) technique was used for detection of membrane fluidity, while biological illumination method for dectection of SOD. RESULT: The membrane lipid fluidity of lymphocytes obtained from broncho-alveolar lavage fluid (BALF) in the pulmonary tuberculosis and lung cancer patients, and the membrane lipid fluidity of carcinoma cells in lung adenocarcinoma and squamous cancer patients were found to be 4.026 +/- 0.722, 38.254 +/- 0.100, 22.557 +/- 3.771 and 32.875 +/- 9.709 respectively, and statistically significant difference was found between pulmonary tuberculosis and lung cancer patients. The contents of SOD were 170.7 mg/L in the pulmonary tuberculosis and 65.4 mg/L in the lung cancer patients, and statistically significant difference was also found between the two groups. The contents of SOD in the mild, moderate and severe tuberculosis patients were 270.8 +/- 4.1 mg/L, 160.2 +/- 1.9 mg/L and 90.4 +/- 1.6 mg/L respectively, and SOD contents decreased with deterioration of clinical status. CONCLUSION: The membrane lipid fluidity and SOD might be useful biological markers for differential diagnosis between pulmonary tuberculosis and lung cancer. PMID- 10374461 TI - [Surgical treatment in pulmonary tuberculosis with sputum long-term Mycobacterium tuberculosis positive]. AB - OBJECTIVE: To evaluate the importance of surgical treatment in pulmonary tuberculosis with sputum long-term Mycobacterium tuberculosis positive. METHOD: Sixty-five patients with pulmonary tuberculosis who failed antituberculosis chemotherapy received lung resection or thoracoplasty. RESULT: Sputum negative conversion was found in sixty-three patients without recurrence during 1-3 year follow-up, while sputum positive conversion was found only in one case during the period of follow-up. Among all the patients, one died of respiratory failure after the operation, three complicated with empyema and recovered from operating again. CONCLUSION: Surgical operation is still important for treating pulmonary tuberculosis patients with sputum long-term Mycobacterium tuberculosis positive. PMID- 10374462 TI - [Relation between pulmonary surfactants and asthma]. PMID- 10374463 TI - [Drug resistance mechanism of doxorubicin-resistant human gastric cancer cells BGC-823/DOX]. AB - To further investigate drug resistance mechanism of BGC-823/DOX, we analysed the amplication, transcription and overexpression of genes associated with drug resistance by Southern blot, RT-PCR, immunohisto clogical assay and the concentration of Doxorubicin in the cells by flow cytometry. The results showed that parent cells of BGC-823 were cell line with intrinsic drug resistance similar to the gastrointestinal tumors. Multidrug resistance was the main mechanism of drug resistance in BGC-823/DOX. There was a significant difference between the doxorubicin-resistant cells line from parent cells with intrinsic drug resistance and from parent cells without intrinsic drug resistance in drug resistance. PMID- 10374464 TI - [The effect of calcium content in bovine cancellous bone as carrier on expression of tumor necrosis factor-alpha gene in mice]. AB - We investigated the expression of tumor necrosis factor-alpha (TNF alpha) gene in the area of implanted calf cancellous bone with different calcium content and the influence of calcium salts on local cellular immunity. The particles of mineralized and partially demineralized bone were implanted in the mouse's muscle pouch, and removed 3, 5, 10 and 20 days after implantation of the bone particles. The specimens were processed for determining the expression of mRNA encoding TNF alpha, which was performed by a nonradioactive in situ hybridization technique. The expression of TNF alpha mRNA was markedly higher in the mineralized bone group than in the partially demineralized bone group (P < 0.01). The positive rate of TNF alpha gene expression was highest by 10 days after implantation. There was a strong hybridization signal localization to the cytoplasm of morphologically identifiable monocytes and multinucleated giant cells. Similar activity was detected in the cytoplasm or nuclei of mesenchymal cells, fibroblasts as well as striated muscle fibers. This finding suggests that the calcium content in calf cancellous bone possesses a significantly stimulative effect on TNF alpha mRNA expression, and calcium salts may be of importance for the modulation of local cellular immunity. PMID- 10374465 TI - [Effects of cytokines on somatostatin in nude mice bearing human renal cell carcinoma]. AB - We studied the relationship between the production of SS and treatment with cytokines and a new method for the treatment of renal cell carcinoma. 4.4 x 10(6)RCC94616 cells were injected subcutaneously into the back of nude mice. Five groups with TNF, IL-2, rIFN, TNF + IL-2, TNF + rIFN and controls were randomly divided according to the mean diameter of experimental tumor. After the last injection of cytokines, 0.5-0.8 ml blood, 1g tumor tissue, para-tissue and normal tissue were havested respectively. Contents of SS were tested by radioimmunoassay. In the treatment groups with cytokines, the concentration of SS was changed, siginificantly increased in the TNF + IL-2 group (P < 0.01). The effect on distribution of SS by cytokines may also be mediated by the regulation of human immunity and antitumor activity. It may be suggested that the method of TNF + IL-2 + SS is best to treat renal cell carcinoma. PMID- 10374466 TI - [The injury of human lung cultured vascular endothelial cells in vitro added to burned skin extracts]. AB - Human lung cultured vascular endothelial cells model was established in vitro. Two kinds of skin extracts (human burned skin extracts, HBSE, and human natural skin extracts, HNSE) were added to cultured endothelial cells. The results showed that HBSE exerted toxic effect on endothelial cells in vitro. There were significantly higher contents of ET-1, NO, TNF-alpha, and abnormal structural changes in EC in the HBSE group. PMID- 10374467 TI - [Biomechanical observation on unstable intertrochanteric fracture fixed by 130 degrees angled-plate]. AB - We examined the stability of unstable intertrochanteric fracture fixed by 130 degrees angled-plate biomechanically, and the stress distribution of the plate. The results showed that after 300 times circle-loading test, the fracture stability was destroied. Stress concentration was observed on the plate, which was aggravated by circle-loading test. These confirmed the inproper biomechanical stature of the plate. Referring to the clinical review, we conclude that using angled-plate to fix unstable intertrochanteric fracture hardly yields satisfactory results, at least, early wight-bearing exercise should not be promoted. PMID- 10374468 TI - [Significance of surgical treatment of bursting fracture with CT estimation analysis of the anatomy of pedicle of vertebral arch]. AB - Using CT analysis of 3-D anatomical data of the pedicle of the vertebral arch as a guideline, we standardized the pedicle screw insertion technique. Fourty-six bursting fracture cases were treated using a AF 3-D pedicle screw system with the standard technique and followed up for over 12 months. Comparision was made with the former non-standard technique performed in 72 cases. The results showed that each screw reached the maximum depth with an accurate angle. Comparision of the former non-standard technique showed a significant difference between the two groups. 46 bursting fracture cases obtained anatomical reduction in 3-D and rigid fixation after follow-up for 12 months. Pedicle screw, inserted through the pedicle anatomical axis can reach the maximum depth, that will achieve best stability and ideal clinical result. CT analysis supplies reliable individual data for standardization of pedicle screw insertion. PMID- 10374470 TI - [Clinical analysis of 7 cases of mesenteric venous thrombosis]. AB - 7 patients with mesenteric venous thrombosis (MVT) were treated in Anzhen Hospital from 1984 to 1996. The patients were all operated on and the diagnosis was confirmed pathologically. Fine patients were cured and 2 died. We discussed the pathophsiology, pathogeny, clinical manifestation, diagnosis and treatment of MVT. Most of the patients presented with a clinical picture of acute abdomen and the diagnosis was made only at laparotomy. Close observation and proper laparotomy should be given to the doubtful cases. To reduce the mortality rate of MVT, we resected the bowel which looks normal in appearance but comprises the mesentery of thrombosis. Adequate anticoagulant therapy should be given as early as possible. PMID- 10374471 TI - [Diagnosis and management of piriform sinus fistulae in children]. AB - In 12 patients with piriform sinus fistula, aged from 1.5 to 10 years all but one patient presented themslves with inflammatory swelling in the region of the left lobe of the thyroid gland. During acute infection, the involvement of the left thyroid lobe was verified by ultrasonography and scintigraphy. Barium swallow examination was the diagnostic finding that all the 11 patients had a left-sided fistula. One patient presented with a painless cyst in the left thyroid lobe. Six patients operated on 4 underwent fistulectomy. In patient the fistula was not found. In the patient who had a cyst in the thyroid lobe, fistulectomy and cystectomy were done. All these patients did well postoperatively without recurrence. Six other patients were followed up without operation with in 1-6 years. Five patients remained asymptomatic. One patient had 3 recurrences during a 2 years follow-up period. Piriform sinus fistula is the most common underlying abnormality in the patients with acute suppurative thyroiditis. A better understanding about this rare branchial anomaly is of clinical importance. PMID- 10374472 TI - [Effect of extended lymph nodes dissection on curative resection of lung cancer]. AB - We evaluated the effect of extended lymph nodes dissection on curative resection in 386 patients with lung cancer surgically treated for 9 years. The extended lymph node dissection of the hilar and mediastinum was performed on the basis of mapping system for mediastinal lymph nodes developed by Naruke. Altogether 2603 groups of lymph nodes were dissected (mean 6.74 groups). The overall metastatic rate was 49.2% (190/386), including simple N1 diseases (43 cases, 11.1%) and N2 diseases (147, 38.1%). The metastatic rate for squamous cell carcinoma, adenocarcinoma, small cell carcinoma and large cell carcinoma was 30.1%, 44.1%, 48.0% and 50.0%, respectively. The lymphatic metastases in lung cancer showed the features of saltatory infiltration and multiple levels of involvement. Cure could be achieved only after extended dissection of intrapulmonary and mediastinal lymph nodes. PMID- 10374469 TI - [Experiences in management of acute necrotic pancreatitis]. AB - To establish a treatment program for patients with acute necrotic pancreatits (ANP), we treated 211 patients with ANP from 1973 to 1995. 122 patients in group A were treated mainly by early operation before 1992 and 75 in group B, by early nonoperativemethod. The results showed that the occurrance of complications (ARDS, renal failure, heart failure, pancreatic abscess, intestinal fistula) was lower than that in the group B (P < 0.01). Also the mortality in the group B (10.67%) was decreased as compared with that (22.95%) in the group A. We suggested that the nonoperative treatment with rigid criterion for ANP might be recommended first. An intensive nonsurgical management of ANP, surgical indications during nonoperative period and complete program of management for ANP were discussed. PMID- 10374473 TI - [Operative treatment of displaced acetabular fractures through the ilioinguinal approach]. AB - To reduce the incidence of ectopic ossification and improve hip joint function after the operative treatment of displaced acetabulum fractures. We surgically treated 12 acetabular fractures which involved anterior column or both anterior and posterior column fractures through the ilioinguinal approach. 10 patients were judged postoperatively to have an anatomic reduction, and 2 had a satisfactory reduction. An average of 3-year follow-up showed excellent results in 8 patients (67%) and good in 4 (33%). Radiographic results indicated excellent results in 8 (67%), patients good in 3 (25%), fair in 1 (8%). The results suggested that the ilioinguinal approach appears to diminish many of the problems associated with utilization of an extrapelvic approach, including the disturbances of the hip abductor system, heterotopic ossification, sciatic palsy, the slow recovery of hip mobility. We conclude that it is not only good for an operative treatment of anterior column fractures but also good for some of posterior column fractures. PMID- 10374474 TI - [Perioperative management of open-chest surgery for cancer patients in advanced stage in aged and with respiratory insufficiency]. AB - For patients with chest cancer who are in advanced stage, aged, or/and with respiratory insufficiency, the key to the success of open-chest surgery is intensive management of the respiratory tract and proper treatment of complications in perioperative period. From May 1984 to November 1995, 1661 patients with esophageal and lung cancer underwent thoracotomy in our hospital. 198 of the 1661 patients fit in with the standards of advanced, aged or respiratory insufficiency. The patients were aged over 70 years, with the length of esophageal cancer over 10 cm and, the MVV less than 50% (least 24.3%). The results the patients were satisfactory because a series of the effective measures of perioperative management were tahen. They incladed thyrocrico-puncture and use of a fine tube through which drugs were injected to irritate cough sputum out; examination and suction with bronchofiberoscope; tracheo-bronchial lavage through bronchofiberoscope with antibiotics solution; and tracheotomy using ventilator appropriately. Perioperative intensive management is very important for extending operation indication and tiding over critical period. PMID- 10374475 TI - [Reoperations on prosthetic heart valves: an analysis of 64 cases]. AB - During the period of Fabruary 1988 to May 1996, 64 reoperations were performed for prosthetic valve malfunction. Degeneration of bioprosthesis occurred in 46 patients, failure of mechanical prosthesis in 9, and periprosthetic leakage without evidence of infection in 9. In accordance with the status of cardiac function, 9 patients with acute dysfunction of mechanical prosthesis received emergency operations, and the remaining 55 patients with bioprosthetic valve failure or periprosthetic leakage received selective operation. There were 8 early deaths with a overall hsopital mortality rate of 12.5%. Three of the deaths occurred among the 55 selective procedures for a mortality of 5.3% and 3 of the deaths occurred among the 9 emergence procedures for mortality of 33.3%. The 56 long-term survivors were followed up from 3 months to 7 years (mean 2.1 years). There were 3 late deaths (5.3%). Forty-eight patients have been living more than 1 year. Their cardiac function after reoperation was class I (42 patients), class II (5), and class III (1). The timing of reoperation and surgical technique were discussed. PMID- 10374476 TI - [Meningo-cerebral arteriovenous malformations]. AB - 22 patients with meningo-cerebral arteriovenous malformations, we treated in our hospital between 1990 and 1995. Clinical manifestations included headache, nausea, seizure, intracranial hemorrhage, and progressive hemispheric neurologic deficits. The niduses were extensive and had dual feeding from internal and external carotid arteries. All patients underwent endovascular treatment and 4 surgical operations. The curative effect was satisfactory. The clinical and angiographic characteristics were discussed. PMID- 10374477 TI - [Clinical significance of residual urine volume in bladder outlet obstruction with benign prostatic hyperplasia]. AB - Bladder outlet obstruction (BOO) with benign prostatic hyperplasia (BPH) was determined by pressure-flow studies. Residual urine volume was measured by urethral catheterization. Fifty-one patients with or without BOO were investigated for residual urine. The mean residual urine volume was 32.8 +/- 35.9 ml and 25.2 +/- 22.7 ml in BOO group (31 patients) and no BOO group (20) respectively. The range of residual urine volume was from 6 to 210 ml and from 3 to 88 ml in the two groups respectively. Residual urine volume was not statistically different between the BOO group and no BOO group (P > 0.05). The results of pressure-flow studies were significantly different between the BOO group and no BOO group (P < 0.01). Residual urine determination was not a reliable criterion for selection of patients for surgery. The cause of residual urine is not only BOO. The patients with BOO may present insignificant residual urine. It is evident that the absence of post-void residual urine does not rule out BOO. PMID- 10374478 TI - [Advances in partial liver transplantation from living donors]. PMID- 10374479 TI - [Hope is placed on younger surgeons in general surgery in China]. PMID- 10374480 TI - [The evaluation of endoscopic biliary drainage for 288 patients with malignant hilar obstruction]. AB - Hilar tumors are extremely difficult to manage with a considerably lower resection rate. We performed endoscopic biliary drainage for 288 patients with hilar tumors (Klatskin tumor 184, gallbladder carcinoma 23, HCC 47 and other metastases 34) in the past 3 years. 162 patients underwent endoscopic nasobiliary drainage, 80 plastic biliary stenting, and 46 expandable metal stent implantation. 4 patients were given double stents insertion simultaneously, 43.1% of patients received good drainage with the total effective rate of 67.0%, but postprocedure cholangitis took place in 13.8% of patients within one month and 3 died of cholangitis and sepsis. In the long-term follow-up patients without surgical treatment, the median sruvival was 5.3 months. The outcome was closely related to Bismuth types, and jaundice could be relieved if more than about 40% of the liver was drained. The double stents for the left and right intrahepatic duct in the meantime could enlarge drainage area and improve the theraputic effectiveness. To get highest benefit, the 3 endoscopic biliary drainage methods should be choosen properly and exchanged flexibly. We conclude that endoscopic biliary drainage is a safe and useful management for the hilar tumor and should be the treatment of choice for palliating jaundice in the inoperable patients. PMID- 10374481 TI - [Choice of time and mode of operation in severe acute pancreatitis]. AB - We report the treatment of 75 patients of severe acute pancreatitis. Operations were performed on patients with pancreatic infection and organ failure according to the principle of "individualization". Operation was also indicated for the patients aged 60 who had had no improvement after a short period of non-operative treatment because of the susceptibility to develop MOF, Non-operative treatment was administered to other patients. This therapeutic mode can increase the cure rate while complications can be decreased. Early operation is necessary for patients with single organ failure in case sequential MOF might happen. The main components of operation were elimination of necrostic lesion, lysis of the pancreas, multiple drainage, and adequate postoperative peritoneal lavage. This operation can not only eliminate necrostic tissue, but also prevent progressive pancreatic necrosis. It introduces free drainage, and can bring satisfactory therapeutic result and reduce the possibility of reoperation. PMID- 10374482 TI - [Diagnosis and treatment of congenital choledochal cyst]. AB - From January 1980 to June 1996, 82 patients with congenital choledochal cysts were treated and 76 of them were operated on B-ultrasonic diagnosis was made with a correct diaginostic rate of 93.9%. The total effective rate was was 78.4% after follow-up for 84 months. Resection of the cyst with Roux-Y hepaticojejunostomy was successful in 96.2% of the patients. Outer drainage was used as the first-aid measure. Cystoduodenostomy and cystojejunostoy which may lead to complications should be abandoned. PMID- 10374483 TI - [Combined laparoscopic surgery]. AB - With minimal and flexible access, combined laparoscopic surgery (CLS) is most suitable to treat several abdominal diseases simultaneously. Among 1000 cases of laparoscopic procedures since 1991, 52 were CLS. They were laparoscopic cholecystectomy (LC) + appendectomy (LA) 24, LC + laparoscopic fenestration of liver cyst (LFLC) 2, LC + laparoscopic herniorraphy (LH) 4, LC + laparoscopic common bile duct exploration (LCBDE) 12, LC + laparoscopic salpingo-ophorectomy 1, LC + laparoscopic removal of leiomyoma of uterus (LRLU) 1, LC + laparoscopic excision of hemangioleiomyolipoma (LEH) in right kidney 1, LH + laparoscopic varicocelectomy (LV) 2, laparoscopic paraesophageal hernia repair + nissen fundoplication 5. CLS was successfully performed laparoscopically without morbidity and mortality. One case of LC + LCBDE and one case of LH + LNF converted to open surgery be cause of unexpected injury of cystic artery and esophageal rupture. Compared with single laparoscopic surgery, CLS did not increase postoperative pain, hospitalization, recovery period, and cost for equipment and instruments. Our results showed that as long as we insist on basic surgical principles, strictly follow operative indications for CLS, more and more patients with multiple abdominal diseases will enjoy minimal access surgery. PMID- 10374484 TI - [The effects of IFN-gamma on the antitumor abilities of immunocytes and chemotherapeutic agents]. AB - We studied the effects of IFN-gamma in immunotherapy and chemoimmunotherapy. The effects of IFN-gamma on TIL and LAK cytotoxicities on K562 tumor cell line and MKN45 gastric cancer tumor were observed. The antitumor effects of 5 chemotherapeutic agents with or without IFN-gamma on SGC7901 gastric cancer cell line were evaluated. The results showed that: (1) IFN-gamma obviously enhanced the cytotoxicities of TIL and LAK on 2 cell line: (2) IFN-gamma also increased the antitumor abilities of 5 agents on SGC7901 cell line. The study suggested that IFN-gamma is useful in immunotherapy and chemoimmunotherapy as an important immunomodulating agent. PMID- 10374485 TI - [Effect of phentolamine on portal pressure in cirrhotic patients with portal hypertension]. AB - Changes of hepatic venous wedge pressure (WHVP) and portal venous pressure (PVP) in fourteen patients with postnecrotic cirrhosis proved pathologically and seven patients with colon carcinoma without hepatopathy were observed before and after the infusion of alpha-adrenergic receptor antagonist phentolamine from peripheral venous. The results indicate that phentolamine can reduce WHVP and PVP of cirrhotic patients with portal hypertension efficiently (0.56-0.63 kPa, 0.42-0.50 kPa, respectively, P < 0.01), but it effects arterial diastolic pressure and heart rate simultaneously (arterial diastolic pressure reduced by 0.94-10.7 kPa, while heart rate increased by 8.67-10.67 per minute). Phentolamine doec not reduce portal pressure on patients without portal hypertension. PMID- 10374486 TI - [Experimental and clinical research of cytolytic T lymphocytes specific for hepatocellular carcinoma]. AB - To investigate a new kind of anti-tumor immunological cells and improve surgical results of hepatocellular carcinoma by anti-recurrence application, we activated T cells isolated from tumor infiltrating lymphocytes by double stimulating signals: the one was autologus HCC cells which were treated with IFN-gamma and TNF-alpha for enhancing expression of MHC class I and presented tumor antigen, and the other was costimulation signals which was from ICAM-1 and B7 molecules expressed on treated HCC cells as well as CD28 mAbs. Activated T cells which bound to HCC cells were expanded selectively as tumor specific cytotoxic T lymphocytes (TS-CTLs), their cytotoxic activity in vitro and anti-tumor effects in vivo were observed. Our results suggested that TS-CTLs expressed high cytotoxicity against autologous HCC cells with MHC class I restriction manner. Adoptive TS-CTLs treatment could decrease serum AFP level, inhibit ascites formation and prolong survival in SCID mice bearing human HCC. In clinical trail of 12 cases of HCC, TS-CTLs treatment was able to delay tumor recurrence after HCC resection. Our data demonstrated that TS-CTLs as new immunological treatment modality, are of great value in further application of tumor comprehensive management. PMID- 10374487 TI - [Molecular mechanism and therapy of hypoalbuminemia in peritoneal infection]. AB - Albumin mRNA expression was studied by the use of reverse transcriptional polymerase chain reaction (RT-PCR) to determine the molecular mechanism in peritoneal infection. Albumin mRNA content markedly decreased. Endotoxin inhibited albumin mRNA expression in vivo probably by stimulating TNF,IL-1, and IL-6 production. Changes of the hormone levels were not the cause of hypoalbuminemia in infection. Reconbined growth hormone and astragalus polysaccharides alleviated inhibition albumin synthesis inhibition increasing albumin serum concentration in rats with peritoneal infection. The results suggested that hypoalbuminemia is associated with endotoxemia during sepsis, which inhibites hepatocytes albumin synthesis probably by stimulating nonparenchymal cells to produce TNF, IL-1, and IL-6. PMID- 10374488 TI - [Influence of recombinant growth hormone on protein metabolism during severe infection: an animal experiment]. AB - We observed the influence of recombinant growth hormone (rGH) on protein metabolism during sepsis and found its mechanisms. Cecal ligation and puncture were choosen to duplicate the severe infection model. Animals of therapy group received rGH 1 U/kg/d after CLP operation, while sepsis group received normal saline. rGH accelerated regaining of the positive nitrogen equilibrium, improved plasma albumin level. rGH accelerated the recovery of intestinal mucosa glutaminase activity, preserved the normal structure of intestinal mucosa, reduced the portal venous endotoxin level and venous TNF level. rGH improved the albumin synthesis of isolated hepatocytes, and inhibited the expression of albumin mRNA level during severe infection. We conelude that rGH preserves the normal structure and function of intestinal mucosa during sepsis, and reduces gut origin hypermetabolism reactions. Moreover, rGH improves the synthesis of protein. PMID- 10374489 TI - [The role of platelet activating factor in pathogenesis of acute pancreatitis in dogs]. AB - 17 Beagle's dogs were divided randomly into tree groups: pancreatitis group (PG, n = 6), pan+BN52021 group (BNG, n = 6), control group (CG, n = 5). The acute pancreatitic model of PG and BNG was established by injecting sodium taurocholate and trypsin into the main pancreatic duct. Animals of BNG were injected PAF receptor antagonist BN52021 (5 ml/kg) intravenously 5 minutes and 3 hours respectively after acute pancreatitis induction. Blood amylase activity was determined by Winslow's method. PAF in blood and pancreatic tissue was determined by the platelet accumulation method. Blood amylase activity of PG increased by 466.7 +/- 111.6 than the baseline at 8 hours and increased significantly than that of BNG and CG (P < 0.05). Blood PAF level of PG increased from 30 minutes to 11.81 +/- 0.78 ng/ml at 8 hours. BN52021 inhibited very significantly the increase of PAF level (P < 0.01). PAF level in pancreatic tissue of PG was significantly higher than that of BNG and CG (P < 0.01). PAF may play an important role in early acute pancreatitis. PMID- 10374490 TI - [The relationship between hemorrhagic dilation of the fourth ventricle and outcome]. AB - 52 adult patients with CT-documented fourth intraventricular hemorrhage (FIVH) were treated. The various etiologic findings included intraparenchymal hemorrhage with secondary FIVH (24 cases), spontaneous subarachnoid hemorrhage (8), spontaneous IVH (12), and trauma (8). Of the 52 patients, 28 died because of hemorrhagic dilation of the fourth ventricle in spite of aggressive therapy. Of the 24 FIVH patients without dilation, 10 died and 14 survived. The survival rate for the patients with hemorrhagic dilation of the fourth ventricle was significantly lower than that for FIVH patients without dilation. Of the 28 patients with FIVH associated with dilation, 24 presented with a Glasgow Coma Scale (GCS) score of less than 5 or 5, 2 with a GCS score of 7, and 2 with a GCS score of 13. In the 24 FIVH patients without dilation, 8 presented with a GCS score of 3 to 5 (seven died and one survived), 4 with a GCS score of 6 to 8 (two died and two had survival), 5 with a GCS score of 9 to 12 (one died and four had survival), and 7 with a GCS score of 13 to 15. There was a great difference of GCS scores between the two groups. Logistic regression multivariate analysis demonstrated that hemorrhagic dilation of the fourth ventricle was the most significant outcome predictor, followed by GCS score and then the presence of diffuse IVH. PMID- 10374491 TI - [Relation of continuous ICP, CPP monitoring with prognosis for severe brain injury]. AB - We analysed the treatment results of two groups of patients. Group I included 50 patients with severe brain injury with GCS 3-8, on whom continuous intracranial pressure (ICP) and cerebral perfusion pressure (CPP) monitoring was performed and Group II included 50 cases of similar patients, on whom no continuous ICP monitoring was performed. In Group I 8 patients had normal ICP (< 2.0 kPa), CPP (> 9.33 kpa), and the rest 42 had increased, ICP and reduced CPP. After adequate intervention including operation and drug treatment, group I patients had good results with a mortality of 14%. Group II patients received the same intervention based on clinical observations, but they had relatively worse results. We are of the opinion that continuous ICP and CPP monitoring for severe brain injury patients helps find proper treatments and reduce mortality. PMID- 10374492 TI - [Endovascular treatment of brain arteriovenous malformation]. AB - Eighteen patients with arteriovenous malformations (AVMs) of the brain were treated with endovascular techniques between 1993 and 1996. The following angiograms showed complete obliteration of the nidus in 2 patients, 70%-90% reduction in nidus volume in 5, 50%-70% reduction in 3, and < 50% in the remaining 3. Two patients suffered from moderate neurologic deficits after embolization. One week after the treatment, 4 patients with superficial AVMs underwent surgery, and 2 with deep-situated AVMs received gamma-knife therapy. 14 patients were followed up, suffered from more frequent epilepsy, and 1 had intracranial haemorrhage 6 months after embolization. A few AVMs can be cured by endovascular embolization. In regard to some large AVMs or AVMs situated at important functional areas, embolization can facilitate surgery and radiotherapy afterwards. PMID- 10374493 TI - [Predictive value of IL-2R in renal tissue during acute renal allograft rejection]. AB - The pattern of intragraft immunological events associated with acute cellular rejection (ACR) has been previously defined. We conducted a retrospective longitudinal study to compare the clinical significance of various immunological markers in renal materials obtained from human renal allografts. Three to 6 consecutive allograft biopsies were obtained from 17 serial human transplant recipients over an 8-month period. A total of 59 core needle biopsies were examined. The expression of interlukin-2 receptor (IL-2R) in interstitial infiltrates, as well as the presence of CD4/CD8 T cell infiltrates were assessed by immunocytochemical staining of frozen sections. When ACR occurred, IL-2R expression increased about 16.3 fold in the renal specimens significantly as compared with those specimens without ACR. After MTX-pulse therapy the expression of IL-2R in biopsies were evidently down-regulated about 60%. The results suggested that the expression of IL-2R in renal allograft may be a useful and helpful adjunct in the diagnosis of ongoing acute cellular rejection, and decrease in these expression may be indicative of successful antirejection therapy. PMID- 10374494 TI - [Measurement of skin area of expansion and defect on scalp]. AB - The skin expander has been widely used to produce extra skin by the plastic surgeon, but there is no a scientific method for measuring the skin area of external expansion. The plastic surgeon usually do not know exactly whether the skin area of expansion more or less than the skin area of defect. A morphologic measure method was developed to figure out accurately the area of expansion and the area of defect separately. Clinical application has proved that the method can help the plastic surgeon know the wanted skin area of expansion in practice before operation. PMID- 10374495 TI - [Application of atracurium to critical care patients in ICU]. AB - Fifty-two patients entered ICU after heart operation with cardiopulmonary bypass. These patients with ventilation atracurium (ATC) were given 3 micrograms-6 micrograms/kg/min for 5-48 hours. No complications happened. After ATC was used, the patients could open eyes, and cooperate with the doctor when they regain consciousness. The circulation index was normal. The limbs of these patients relaxed. In the 52 cases, 30 did't use sedatives, and 22 used senall dose of sedatives for sleep at night. We conclude that ATC does not require hepatic or renal function for termination of effect and action is terminated by the "Hoffman elimination" the ester hydrolysis. The time of effect is short and there is no cumulation effect. There is no vagal or ganglionic blocking activity, so ATC does not influence circulation and myocardial function. Maintenance relaxation keeps ventilation steady and oxygen supply normal. PMID- 10374496 TI - [Emphasis on basic and clinical study on central nervous tissue transplantation]. PMID- 10374497 TI - [Intracerebral grafting of ex vivo transgene muscle cells in rat model of parkinsonism]. AB - The rat model with a unilateral 6-OHDA-induced lesion of the dopaminergic system was used for experimental study on gene therapy for Parkinson's disease (PD). The plasmid pCMVTH (6.04 kb) was constructed as a vector containing the gene of tyrosine hydroxylase (TH) and transferred into cultured primary muscle cells by lipofection ex vivo. After expression of TH was determined in vitro, the intracerebral grafting (ICG) of these genetically modified cells was made in the striatum of PD rat model. The results showed that TH-expressing muscle cells had a long-term survival in the brain and induced a marked decrease in abnormal locomotion and increase in striatal dopamine levels for rat model. PMID- 10374498 TI - [Intracerebral transplantation of genetically modified cells for Parkinson's disease]. AB - The clinical application of brain transplantation for treating Parkinson's disease (PD) is limited because it is difficult to obtain ideal donor cells. In this study, the tyrosine hydroxylase (TH) gene was introduced into NIH-3T3 cell line and the genetically modified cells reacted positively with TH antiserum and released L-dopa. These cells were implanted into the striatum of PD rats. 3,6,9,12 days after implantation, the average rate of the abnormal rotational behavior recovery was 52.6%, 68.4%, 63.2%, 44.7%, respectively. This result is considered to be potential for further clinical application. PMID- 10374499 TI - [Long-term survival of fetal substantia nigra grafts in striatum of PD rats: an observation of tyrosine hydroxylase immunohistochemistry and western blot]. AB - By using immunohistochemistry and Western blot, we observed the varied patterns of survival, development and growth of dopaminergic neurons after intrastriatal transplantation of fetal substantia nigra. There was a phenomenon of "developmental delay" of grafts in the host. As the time of transplantation went on, the grafted dopaminergic neurons could reinnervate the host neurons. The observation reveals that the reinnervation between graft and host is more beneficial to the functioning of the grafts. PMID- 10374500 TI - [Immunological rejection in rat embryonic brain tissue transplantation: experimental study]. AB - The embryonic cerebral neocortex from rat or mouse donors which were 16 to 18-day old embryos was grafted into the neocortex of adult recipient rats. One group was treated with cyclosporin A. At different time after transplantation, the brain graft tissues were stained immunocytochemically to exmine the expression of MHC class II antigens and both subsets of T-cells, helper-inducer and cytotoxic suppressor T-cells. The results indicated that the survival rate of treated group was higher than that of non treated with immunosuppressant. Immunocytochemical evaluation showed that a large number of helper T-cells and cytotoxic T-cells appeared, with a significant increase of the number of class II major histocompatibility complex (MHC) expressing cells within and around the allografts was observed. These MHC-class II antigen-positive cells may be lymphocytes, microglia and astrocytes. We conclude that the CNS is not a "immunologically privileged site" and there is graft rejection in brain transplantation. It is necessary for brain transplantation to be treated with immunosuppressant. PMID- 10374501 TI - [Intraparenchymal and intraventricular transplantation of fetal substantia nigra cell suspension in rat model of Parkinson's disease]. AB - The grafts of fetal substantia cell suspension grafts could bring into a behavioral effect and characteristics of histology and immunocytochemistry in rat models of PD after the grafts were transplanted either into the lateral ventricles or directly into the striatum. It was found that the intraparenchymal tromsplantation of fetal nigral cell suspensions not only was beneficial to reinnervation between host and grafts, but also produced more complete and steady effects than the intraventricular transplantation. We concluded that the restoration of nervous functional deficits of PD rats is likely to depend on both extensive dopaminergic reinnervation throughout the grafted sites and on closer integration of the grafts with the host striatal circuitry. PMID- 10374503 TI - [Hepaticocholangiochole cystostomy plus construction of subcutaneous tunnel of the gallbladder in the treatment of hepatolithiasis with bile duct stricture]. AB - A novel operation was designed by using Hartmann's pouch of the gallbladder to anastomose stone-strictured intrahepatic bile duct while leaving the common bile duct and Oddis sphincter intact and fixing the fundus of the bladder subcutaneously. 28 patients with intrahepatic bile duct stones and strictures underwent the operation successfully, and recovered well. There were no postoperative complications such as reflux cholangitis or disturbances of GI tract. PMID- 10374502 TI - [The repair of injured brain by implanting embryonic cortical brain tissue preserved in Iloprost solution at 4 degrees C: experimental study]. AB - We transplanted the fresh and Iloprost or saline preserved (3 hours) embryonic rat cortical brain tissue into the rats in which the motor cortex had been injured and compared the neurobehavioral and morphological changes. The neurobehavior of the transplanted groups was more significantly improved than that in the injured, untransplanted group. The neurobehavior of the animals of the fresh tissue group and Iloprost preserved group recovered to the preinjurylevel. The survival rate of the Iloprost preserved grafts was 90%, which was higher than that of the saline preserved grafts (P < 0.05). In comparison with the fresh grafts, however, there was no significant difference (P > 0.05). In addition, the growth volume of the Iloprost preserved grafts was more markedly increased than that of the fresh grafts (P < 0.01). We conclude that injured brain may be repaired effectively by transplantation of fetal brain cortex. The Iloprost solution not only maintains of vitality the grafts, but also promotes its survival and growth in the host brain. PMID- 10374504 TI - [Splenic artery embolization for the treatment of traumatic splenic rupture]. AB - We used splenic artery gelfoam embolization to save the spleen Seldinger's method in 20 patients with traumatic rupture of the spleen. All patients had the spleen reserved by single embolizing hemostasis satisfactorily. We delineate the method, clinical value, indication and experience of embolism-hemostasis in detail. PMID- 10374505 TI - [Hodgkin's disease with concurrent infection of toxoplasmosis]. AB - Hodgkin's disease (HD) is a specific type of malignant lymphoma characteristic of local and general lymphadenectasis. Aquired toxoplasmosis (AT) is one kind of lymphoadenopathy without fever and fatigue. When the two diseases coexist, clinical and pathological misdiagnosis may be made. This is the first male case of toxoplasmosis and Hodgkin's disease in China, diagnosed by surgical removal of the major part of the cervical and supraclavicular masses, detection of blood anti-toxoplasma gondii antibody, PCR analysis of toxoplasma gondii DNA, and pathological, ultrastructural and immunohistochemical studies of the tumour tissues. The patient treated by radiation and chemotherapy was abated. PMID- 10374506 TI - [Impingement syndrome of the subacromial area: analysis of 42 cases]. AB - To create a new classification of subacromial impingement phenomena according to different etiology, we treated 42 cases of impingement syndrome. They were 29 male, and 23 females, with an average age of 45 years. 29 patients had typical subacromial pain are signs. Positive impingement test was found in 30 patients. Roentgenographic findings showed that the high density of the great tuberosity (GT) in 27 cases, 8 cases got the cystic changes around the CT, and exostosis formed on the G. T. in 3 cases. The abnormal acromion with sloped or over hanging form was found in 6 cases. 11 cases had an irregular inferior surface of the acromion in two cases with the spur formed at inferior surface of acromion. Four patients combined with calcium deposit at subacromial space. Space narrowing of the AH interval was in noted 12 cases. According to the X-ray findings, 35 cases were classified into the abnormal anatomy structure group. The other 7 patients were in dynamic impingement group. 13 cases of impingement syndrome combined with the complication of rotator cuff tear were treated by the Mclaughlin's procedure for repairing the rotator cuff with the acromion plasty. 11 patients were followed up for more than 6 months after operation. 7 cases had excellent results and 4 cases good results. This classification includes two kinds of impingement syndromes: anatomical of structural and dynamic or dysfunction. PMID- 10374507 TI - [Renal subcapsular effusion cured by using greater omentum packed]. AB - The fistula of proximal convoluted tubule which causes the persistent leakage of the glomerular filtrate and brings about renal subcapsular effusion is very seldom seen. There has been no similar report about it. Up to now, there has been no definite diagnosis and treatment for this disease. We made the advantage of the great absorption power of the greater omentum blood vessels networks to make internal drainage. In this way, the glomerular filtrate was reabsorbed and thus the disease was cured. PMID- 10374508 TI - [Conventional and ambulatory urodynamic study on patients with chronic prostatitis syndrome]. AB - To study the urodynamic findings of chronic prostatitis syndrome (CPS), 58 patients with chronic prostatitis and prostatodynia were subjected to the urodynamic investigation that included conventional medium filling urodynamic (CMG) tests (urethral pressure profile, filling cystometry, voiding pressure-flow study) and ambulatory urodynamic monitoring (AM) by using Dantec Menuet and Urodec 500 AM system. 55 BPH patients and 20 normal men were taken as control groups. CPS patients had function bladder outflow obstruction (FBOO). FBOO resulted in the detrusor instable (DIS, 12.1%), detrusor-sphincter dysnergia (DSD, 100%) and detrusor dysconstractility (DDC, 6.9%), and resulted in clinical symptoms. FBOO resulted from the urethal sphincter spasm and DSD. PMID- 10374509 TI - [Clinical study of the effect of early enteral feeding on reducing hypermetabolism after server burns]. AB - Hypermetabolism of burn patients is characterized by massive loss of lean body mass and obvious decrease of resistance. We evaluated the clinical effect of early enteral feeding on its modulating hypermetabolism in severe burn patients. The REEs (resting energy expenditure) decreased by 27.5%, 29.3%, 24.4% (average 27%) on PBD4, 8, 14 respectively in EFg (early feeding group) than those of DFg (delayed feeding group) (P < 0.05-0.01). To compare with the normal value, the mean REE was 138.8% in EFg, and 160% in DFg, the difference between two groups is significant (P < 0.05). Plasma glucagon, cortisol and urinary catecholamine markedly lowered and serum insulin obviously increased in EFg at most timepoints as compared with DFg (P < 0.05-0.01). It is indicated that early enteral feeding could reduce hypermetabolism after severe burn injury. PMID- 10374510 TI - [Interaction of octapeptide of cholecystokinin, vasoactive intestinal peptide and substance P on dynamics of biliary system and cardiovascular system]. AB - We tested in vivo: (a) the effect of an i.v. infusion of cholecystokinin octapeptide, vasoactive intestinal peptide or substance P on dynamics of biliary system and cardiovascular system, (b) the relation of dynamics of biliary system and cardiovascular system. In 91 anesthetized guinea pigs, left ventricle motility of heart, sphincter of Oddi motility and common bile duct pressure were monitored during the intravenous administration of cholecystokinin octapeptide (CCK-OP), vasoactive intestinal peptide (VIP), substance P (SP) and combination of CCK-OP and VIP. Intravenous CCK-OP increased fasting Oddis sphincter motility index, decreased the basal pressure in sphincter of Oddi, increased common bile duct pressure, and decreased the left ventricle of heart motility. VIP alone showed no significant effect on biliary system and cardiovascular system, but in conjunction with CCK-OP it produced inhibition on the effects of CCK-OP on both sides. Exogenous SP acted like CCK-OP on both biliary system and cardiovascular system, but it was less potent. We conclude that it may be an important interaction between dynamics of biliary system and cardiovascular system; and gastrointestinal peptide plays an important role in this interaction in guinea pigs. PMID- 10374511 TI - [Establishment of a new rat model of portal hypertension by intraportal injection of microspheres]. AB - Microspheres were injected intraportally to block intrahepatic portal radicals and to produce a new rat model of portal hypertention. Different sized microspheres (15 microns, 40 microns, 80 microns) were injected into the portal vein. The resultant changes in arterial, portal, hepatic venous and splenic pulp pressures were monitored. The results showed that a small-dose injection of 80 microns microspheres (1.8 x 10(5)) produced a steady state portal venous pressure of 2.53 +/- 0.17 kPa, and all rats showed a normal arterial pressure. This volume of 80 microns microspheres was considered as the suitable dose for the induction of portal hypertention in the model. Besides, numerous microspheres were found in the pulmonary vascular bed, suggesting the existence of intrahepatic portal systemic shunts in the normal rat liver. PMID- 10374514 TI - [Pancreatic surgery improvements in China]. PMID- 10374512 TI - [An experiment study of reversed pulmonary hypertension with inhaled nitric oxide on smoke inhalation injury]. AB - We evaluated the effect and mechanisms of reversed pulmonary hypertension with inhaled nitric oxide (NO) on smoke inhalation injury in the dog model, 21 dogs were divided into 3 groups randomly. Following smoke inhalation, the control group (n = 8) inhaled O2 (FiO2, 0.45) and the treated group (n = 9) inhaled O2 and 0.0045% (45 ppm) NO. Hemodynamics was serially measured for 12 hours. In addition, 4 dogs without smoke inhalation were used to study the normal lung histomophologic findings. The data were analyzed by ANOVA. After inhalation of NO, the mean pulmonary artery pressure (mPAP), pulmonary minute vessels pressure (Pmv), and pulmonary vascular resistance (PVR) were decreased significantly (P < 0.05), while the mean aortic pressure (mAP) and total peripheral resistance (TRP) were not remarkably changed (P > 0.05). The levels of cyclic guanosine monophosphate (cGMP) were increased significantly (P < 0.01). Inhaled NO may reverse pulmonary hypertension with smoke inhalation injury in dogs. The mechanism of selective pulmonary circulation was increased cGMP level in smooth cells. Inhaled NO may be recommended for clinical application. PMID- 10374513 TI - [Fas system in the study of neoplasms]. PMID- 10374515 TI - [A 20-year experience in surgical management of acute necrotizing pancreatitis]. AB - From 1974 to 1994, 243 patients with acute necrotizing pancreatitis (ANP) proved by operation or CT scanning system were treated. The overall survival rate was 70.4%. In 1974-1987, the resection of necrosis was radical and as early as possible, and the main type of operation was subtotal pancreatectomy. The survival rate in this period was 61.3% (49/80). In 1988-1991, the treatment was changed into "individualization" i.e. for sterile pancreatic necrosis, conservative treatment was setected while infected pancreatic necrosis, necrosectomy was performed early. In this period, the survival rate of conservative treatment was 85.7% (6/7) and that of operative treatment 67.1%. The overall survival rate in was 68.5% (63/92). In 1991-1994, we insisted on late operation for the patients with necrosis and comprehensive management for all the patients with ANP. The survival rate of conservative treatment in this period was 100% (11/11) and that of operative treatment 80% (48/60). The overall survival rate was 83.1% (59/71). We conclude that comprehensive management combined with late operation for the patients with necrosis infected is a better strategy for the treatment of ANP. PMID- 10374516 TI - [Indications for operation in patients with acute necrotizing pancreatitis]. AB - The opportunity of and indications for operation in patients with acute necrotizing pancreatitis (ANP) are still controversial. One hundred and ninty surgical cases of ANP were analyzed. Early operation was performed in 2 weeks from onset, and late operation after 2 weeks. In the early operation, the pancreatic sporadic focal necrosis was found in most cases (75.6%), and main postoperative complications were disturbance of blood circulation and organic function injury of pancreas. The postoperative mortality rate was 28.2%. In the late operation, extensive fusional pancreatic necrosis was found in most cases (53.7), and most complications were located in the organs mear the pancreas. The authors conclude that late operation is better for the treatment of ANP. The indications of early and late operation were discussed. PMID- 10374517 TI - [Early diagnosis of pancreatic infection by CT-guided needle-aspiration]. AB - 35 CT-guided percutaneous needle-aspirations with bacteriological sampling were performed in 21 patients suspected of pancreatic infection out of 122 patients with severe acute pancreatitis. Thirteen patients were found by smear and staining culture to have pancreatic infection, and diagnosis was confirmed by surgery. The pathogens found were E coli, pseudomonas and fungi. All pancreatic infections were in grade D or E according to the grading system of Balthazar. Seven of 8 patients with sterile aspirates were cured by nonsurgical therapy. We conclude that CT-guided aspiration is a safe, accurate method for identifying infection of the pancreas at the early stage of severe acute pancreatitis. PMID- 10374518 TI - [Clinical analysis of 153 pancreaticoduodenectomies]. AB - 153 pancreaticoduodenectomies performed from 1976 to 1995 were analyzed. The morbidity was 41.2% and the mortality 5.2%. Delayed gastric emptying (DGE) was the most common complication, and severe complications included introabdominal abscess, upper GI bleeding, and pancreatic fistula. The mortality decreased significantly from 15.6% before 1990 to 0.9% after 1990 and the morbidity decreased from 44.4% to 31.9%, and delayed gastric emptying was the major cause of mobidity after 1990 in pylorus preserving procedures. PMID- 10374519 TI - [En bloc vein resection in the treatment of pancreatic tumors adherent to the superior mesenteric-portal vein confluence]. AB - Contraindication is often considered to pancreaticoduodenectomy for patients with malignant tumors of the pancreatic head. From 1989 through 1995, 15 patients with tumors of the pancreatic head adherent to the superior mesenteric-portal vein confluence (SMPV) underwent pancreaticoduodenectomy with en bloc resection of SMPV. There was no perioperative mortality. Follow-up data showed a mean survival of 17.1 months among 7 deaths, which was significantly longer than an average of 3.8 months among previous 17 patients who abandoned radical resection (P < 0.001). We consider that pancreaticoduodenectomy with en bloc resection of SMPV is feasible in some cases. PMID- 10374520 TI - [Combined resection of the pancreas and portal vein replaced by blood vessel prosthesis for pancreatic head carcinoma: report of two cases]. AB - The resectability and prognosis of carcinoma of the pancreatic head are still poor. The higher resectability rate was achieved by combined resection of the pancreas and portal vein. Two patients in whom the portal vein, surrounded but not penetrated by pancreatic carcinoma, was resected and replaced by an expanded Teflon (Gore-Tex) tube (diameter 8 mm). One patient, who survived 32 months without recurrence of disease, had a ultrasonography and CT proved patent graft 6 and 32 months after operation. A second survivor had a patent graft at 20 month. Excision of involved portal vein during pancreatoduodenectomy for carcinoma of the head of the pancreas is a feasible and valid procedure in the absence of metastases. Vein grafting is the best means for portal vein reconstruction. Gore Tex appears to be suitable prosthesis when the portal vein must be sacrificed. PMID- 10374521 TI - [Pancreatic ischemia: a continuous injury factor in acute necrotic pancreatitis]. AB - ANP model in rats was used to determine the concentration of TXB2, PGF1 alpha in plasma and the ACE activity in serum in five groups. The concentration of TXB2, PGF1 alpha in all experimental groups was significantly different from that of control group (P < 0.05). The comparison of ACE activity was just the same as the above except that of 6 h. The factors leading to pancreatic ischemia functioned continously. We conclude that pancreatic ischemia is a continuous injury factor in ANP, and there is no reperfusion-injury in the course of the disease. PMID- 10374522 TI - [The proliferation inhibition and differentiation inducing effects of all-trans retinoic acid on human pancreatic adenocarcinoma cell line JF-305]. AB - We detected the antiproliferative effect with MTT test and investigated the changes in biological properties, cytomorphology and ultrastructure through cytopathology and electronic microscopy. Cell growth of JF-305 was inhibited by all-trans retinoic acid (ATRA). The maximal inhibitory rate was 34.7%. The number of proliferative cells reduced (P < 0.01). Cell metabolism slowed down, secretory functions recovered, and malignant degree decreased. ATRA can inhibit the proliferation and induce the differentiation of human pancreatic adenocarcinoma JF-305 cells. PMID- 10374523 TI - [Surgical treatment of optic neuropathy after head trauma]. AB - We studied the surgical treatment of patients with optic nerve injury following craniocerebral trauma. Fifteen patients aged 17 to 42 years sustained optic neuropathy after head trauma. Single lateral frontal injury occurred in 11 patients, bilateral frontal injury in 3 and temporal injury in one. After head trauma, 8 patients with visual acuity impairment had no light perception, including 3 patients who had bilateral eyelosing light perception, 5 patients who had only light perception, one who patient can count fingers. In addition, 2 patients compromised supraorbital fissure syndrome, 3 patients complicated exophthaloms, and one patients did enophthaloms. Thirteen patients underwent frontal craniotomy with bicoronal skin flap. Operative procedure obtained the lift of depressed orbital fragment, debridement of hemorrhage, and stabilization of displacement bone. The suprawall of optic canal was opened with microb drill, and longitudinal incision was performed in relatively avascular regions of the optic nerve sheath. Lateral orbital wall approach through peritoneal incision entered the orbital apex in 2 patients with supraorbital fissure syndrome, in who in decompression of suprorbital fissure and optic nervous canal were completed. For the purpose of analysis, the outcome of improvements of visual acuity after surgery was classified into five grades: no light perception, light perception, hand motion, counting fingers, and seeing acuity chart. Visual acuity improvements reaching 2 grade or more were determined as effective, and less than 2 grade as inefficient. All patients were followed up for intervals of 1 month, 3 months, and 6 months. Ten of 15 patients, who had no postoperative complication, showed the improvement of their visual acuity. Exophathaloms was recovered in 3 patients, and so did in one patient enophthalmos. Transcranial approach may be a safe and effective treatment for the treatment of traumatic optic neuropathy in the orbital, intracranial, and optic canal segments after head trauma. PMID- 10374524 TI - [Characteristics of diagnosis and treatment of traumatic intracerebellar hemorrhage]. AB - We treated 10 cases of traumatic intracerebellar hamorrhage, which accounted for 3.7% of intracranial hemnatomas patients in the same period. Two cases had an initial GCS score less than 8. and 8 patients had GCS score of 11-15. Hematomas averaged 6.39 ml. It's a major method for analysis of traumatic mechanism and CT scan. Under normal conditions surgical operation is necessary. The hemispheric hematomas of limited size (less than 3 cm) may be treated conservatively when consciousness is stable. PMID- 10374525 TI - [Pathogenesis and combined treatment of sterility in men with varicocele]. AB - To investigate the pathogenesis of sterility in men with varicocele and summarize the effects of combined treatment, we anallyed ultrastructure of testicular specimens in 21 cases. With combined surgical and medicinal treatment, 16 men (76.2%) made their wives pregnant during a period of 2 years after the operation. Electron microscopy disclosed glycogen infiltration of sertoli cells, and decreased spermatogenesis especially involving the middle and late stages. Defective carbohydrate and energy metabolism in the testis is suggested as the key point in the pathogenesis of sterility, and a combined surgical and medicinal treatment is beneficial to the increase of pregnant rate. PMID- 10374526 TI - [The diagnosis and surgical treatment in rectal endometriosis]. AB - From Jun 1975 to Mar 1995, 26 cases of endometriosis in the rectum (RE) were admitted. Local resection was performed in 16 and Dixon's operation in 10. The result showed 21 cases were symptomatically cured and 5 with remission. 2 cases were reoperated because of recurrence in 5 years after the first operation and cured. RE is difficult in distiguishing from rectal cancer. It is characterized by tenesmus, bursting pain, hematochezia in young and middle aged women during periods. The diagnosis can be made by tipycal history and vaginal examination, rectal examination, barium enema, proctoscopy and so on. The indications of operation include severe clinic symptoms and failed conservative therapy. Wedge resection was suitable in cases with small lession in rectum, while large, deep seated lessions in lower rectum were treated with Dixon's operation in order to prevent recurrence. PMID- 10374527 TI - [Qualitative grading of detrusor contractility in benign prostatic hyperplasia patients]. AB - To grade qualitatively detrusor contractility in benign prostatic hyperplasia (BPH), we studied 120 BPH patients aged from 58 to 89 years. The pressure-flow studies were performed. The passive urethral resistance relation (PURR) and Schafer diagram were used to analyse the results. The detrusor contractility was divided into 4 classes: very weak (VW), weak (W), normal (N), and strong (ST). The types of pressure-flow were high pressure--low flow (HP-LF), high pressure- high flow (HP-HF), low pressure--low flow (LP-LF) and low pressure--high flow (LP HF) type. The results showed that the incidence of VW, W, N and ST class was 12.5%, 26.7%, 47.5% and 13.3% respectively; the incidence of HP-LF, HP-HF and LP LF type was 55%, 1.7% and 43.3% respectively. The weak detrusor strength could become powerful by the surgery treatment of bladder outflow obstruction (BOO) and the function training of detrusor. The detrusor strength class of a BPH patient was improved from W before surgery to N after surgery in our studies. The principles of the fluid dynamics and urodynamic and the clinical application of the qualitative grading of detrusor contractility were discussed. PMID- 10374528 TI - [DNA content correlation with the prognosis of pheochromocytoma: 10 cases report]. AB - We investigated the relationship between DNA ploidy and the prognosis of pheochromocytoma. DNA ploidy studies on paraffin-embedded archival specimens from ten cases of pheochromocytoma were performed by flow cytometry according to Hedley's method. Clinical features and pathological results were investigated and follow-up was made. The values of DNA content in assessment of the prognosis of the tumor were discussed. Flow cytometry examination showed that seven cases had aneuploid, two deploid and one tetraploid. The malignent case was tetraploid, and died from matastasis to the bone. The other cases had a benign process. The examination of DNA content is of value in the assessment of the prognosis of pheochromocytoma. PMID- 10374529 TI - [Detection of p53 and MDM2 gene expression in osteosarcoma with biotin-labelled in situ]. AB - We studied the mRNA expression of p53 and MDM2 gene in osteosarcoma. Twenty osteosarcomas were examined with biotin-labelled oligodeoxynucleotide probe in situ. Of 20 tumors, 8 (40%) and 6(30%) showed positive expression of p53 and MDM2 genes, respectively. Among these, 2 tumors revealed co-expression of both p53 and MDM2 genes. This study confirmed the existence of aberrant mRNA expression of p53 and MDM2 genes in osteosarcoma. PMID- 10374530 TI - [The actions of nucleating proteins in vesicle aggregation and fusion: a preliminary study]. AB - After investigation of the effects of Con A-binding proteins, the major biliary pronucleating effectors system on the figures and lipid composition of model biliary vesicles, the authors found that Con A-binding proteins accelerate aggregation and fusion of vesicles as a result of increased vesicular cholesterol and decreased vesicular phospholipids as well as cholesterol-saturated vesicles. We also found that vesicular proteins though in small quantity are potent in pronucleating effects. The distribution of several known nucleating proteins between the vesicles and micelles is quite different, and there are higher content and more potent pronucleating effectors in patients with cholesterol gallstone than those with pigment stone. By affinity staining method, con A binding proteins were shown in native biliary vesicles as lipid-protein complex. These results suggest that the existence and changes in their quantity and quality of vesicular nucleating proteins play an important role in vesicle aggregation and fusion. PMID- 10374531 TI - [Influence of fibrin glue on biliointestinal anastomotic scar formation]. AB - Conventional suture technique in the reconstruction of the bile duct often result in anastomotic stenosis and eventual obstructive jaundice and biliary cirrhosis. To overcome the aforementioned drawbacks, fibrin glue (FG) was used instead in the biliointestinal anastomosis where indwelling stent was inserted to complete the experimental model. The experiment consisted of pathological study and ECVD evaluation of anastomotic specimens in order to follow up the whole process of anastomotic wound healing. The experimental studies dis-closed that FG promotes wound healing, limits scar formation, and accelerates scar softening and maturnation. FG plus 6-month indwelling stent can guarantee scar maturation. It is an effective method in the prevention of bile duct stricture. PMID- 10374532 TI - [Evaluation and repair of bladder function after spinal cord injuries]. PMID- 10374533 TI - [Make great efforts to raise therapeutic effects of metastatic bone tumors]. PMID- 10374534 TI - [Microwave heating and neoadjuvant chemotherapy for malignant bone tumor]. AB - Limb salvage is widely used but has different disadvantages. Attempting to establish a new limb conservative procedure, we treate patients with malignant bone tumor by intraoperative microwave heating on tumor bearing bone in place and neoadjvant chemotherapy. The tumors were localized at distal femur in 16, proximal tibia in 5, femur shaft in 2, and illia in 2. Pathological diagnosis showed osteosarcoma in 16 patents, parosteoal osteosarcoma in 4, chondrosarcoma in 3, and leiomyosarcoma in 2. The stage of tumor was II b in 18 patients, II a in 6, and III b in 3. Operative technique was wide resection for tumor in soft tissue, protecting the surrounding tissue from heat injury by copper net, and cheating tumor bone at 50 C for 15 minutes. Follow-up varied from 4 to 180 months (mean 57 months). The 5-, 10-years surviral rates were 70.19% and 54.83% respectively. Functional results of 21 patients were > 15 points by Enneking system. The advantages of the method and keeping the shape and continuity of the tumor bearing bone without osteotomy and internal fixation, controlling the time and temperature of heating treatment easily, benefiting for bone remodeling of heated bone. The indications, complications, and surgical procedure of this method were discussed in detail. PMID- 10374535 TI - [Limb salvage surgery using massive allografts in malignant bone tumors]. AB - The advantages of segmental allografts over arthoplastics are, among others, secured biological healing, easy attachment of tendons and ligaments, and desirable long-tem results. Since 1983, the authors have performed 35 resections of malignant tumors in the extremities followed by reconstructions with implantation of segmental allografts. Five types of grafts were used for the reconstruction of the skeletal defects, intercalary hemicylinder, intercalary cylinder, partial osteoarticular, total osteoarticular, and allograft-prosthetic implant composite. All the patients underwent adjunctive chemotherapy that was considered necessary. The length of follow-up ranged from 6 months to 10 years during which time 6 patients died, 25 were disease-free, and 4 survived as tumors carriers. Complications in 28.6% of the patients included infection, graft resorption, recurrence and nonunion. Patients undergoing intercalary grafts oblained better functional results than those having osteoarticular grafts. 4 patients had biopsies and bone scans of the grafts 2-4 years postoperation with creeping substituion being observed. Segmental allograft remains a viable approach with predictable results for reconstruction of skeletal defects after resection of malignant bone tumors. PMID- 10374536 TI - [Pathological type of liposarcoma and its effects of clinical treatment]. AB - To study the pathological type of liposarcoma and it effects of clinical treatment, reviewed 21 cases of liposarcoma. Since Oct. 1976 to Dec. 1995, 21 cases of liposarcoma were treated by operation. 15 cases were male and 6 cases female. The onset of the disease occurred between 6-79 years of age with an average of 49.9 years. 13 cases were well-differentiated, 4 Myxoid, 4 cases of andifferentiaed. All cases were followed up with an average of 8.9 years. The 5-, 10-year survival rate was 94%, the 5-, 10-year unrecurernce rate was 72% and 52%. Among the malignant soft tissue tumors liposarcoma, was most common. The survival rate was related to the pathological type, the site and the size. Good clinical results were well-differentiated and myxoid. The poor was retroperitoneal liposarcoma. The smaller the tumor, the better the result. For the treatment, wide margin excision is a good method of choice. PMID- 10374537 TI - [Panniculitis ossificans and review of the literature]. AB - Four cases of panniculitis ossificans were treated in 1978-1994. They all occurred at the distal phalanx. Pain, swell beneath nails, ulcer, pyorrhea, clark red skin were observed. X-ray examination of 2 cases showed dense shadows in the solft tissue of the distal phalanx. Pathologically the lesion was flatten-round, and separated with the bone. The cut surface was gray and sandy. Histological pictures were the same in 4 cases, namely atrophy of subcutaneons adipose tissue, the osteoid and new bone were seen. Two cases formed the cartilage and the background showed small vessels, proliferative fibroblasts, and inflammatory cells. This disease is easy to recur when nail extraction with curretting is done. It is usually misdiagnosed as inflammation of nail bed or tumor. PMID- 10374538 TI - [Endoscopic reconstruction of 192 patients with traumatic urethral stenosis]. AB - We treated traumatic urethral stenosis with endoscopic technique and observed effects. 192 men with traumatic urethral stenosis (47 with anterior urethral stenosis and 145 with posterior urethral stenosis) were treated with hydroelectric shock wave, cold knife, microwave and electroincission through transurethral endoscopy. We punched hole in the centre of the scar with hydroelectric shock waves for long segment urethral complete occlution. 43 patients were treated with obliteration and 50 with endoscopic operation again for 1-3 times. With a mean of 29. 3 months (range 3-60 months) of followup, 187 patients were treated successfully. Endoscopic operation has good effects on traumatic urethral stenosis with smaller trauma, and few complications. PMID- 10374540 TI - [A self-designed crooked suturing needle anastomosis in the treatment of posterior urethral strictures or occlusions]. AB - A self-designed crooked suturing needle was described and has been used in the surgical treatment of complicated posttraumatic posterior urethra strictures or occlusions in 27 cases from Mar. 1986 to Feb. 1996. 1-stage transperineal end to end anastomosis of the bulbomenbranous urethra to the prostatic urethra with indeed healthy mucosa to healthy mucosa after scar tissue had been excised was performed with the above mentioned crooked suturing needle. 27 patients were followed up for 3 months to 9.5 years after operation with a satisfactory result of 100% and all voided freely with no sound urethral dilation and no trouble after the removal of the catheter. The erect dysfunction of 5 of the patients after the operation within 1-5 years restored completely. PMID- 10374539 TI - [Operative treatment of vesical diverticula]. AB - We reviewed the results of operative management of 31 patients with vesical deverticula, and introduce a simple technique for treating large bladder diverticula. In the 31 patients with vesical diverticula analysed, 23 were followed up for at least six months. Of the 31 patients, 25 were male and 6 female. The average age was 54.2 years. Diverticula was secondary to bladderoutlet obstruction. Seven cases had intradiverticular tumors, and 9 stones in the sac. The diagnosis of vesical diverticula was usually made by cystogram or ultrasonography. The operative indications for vesical diverticula included stone formation, intradiverticular tumor, ureteral obstruction, incomplete empting diverticulum and urine retention due to large diverticula. The combined extravesical and intravesical method was often used. 26 patients underwent both operations for outlet obstruction and vesical diverticula at the same time. Of the 23 followed-up cases, 21 had no symptoms of the urinary tract. Five of the 7 patients with intradiverticular tumor died within 2.5 years; one of the rest survived for 2 years, and the others for 6 years. Two patients with large vesical diverticula received intravesical separation of diverticula. No complications encountered in this simple, time-saving and safe procedure. Cystograms revealed normal condition. The choice of operative techniques to treat vesical diverticula varies with each individual patient. The technique of intravesical separation of vesical diverticula is suitable for large, adhesive and posterior diverticula. PMID- 10374541 TI - [Surgical correction of penile torsion]. AB - To treat the physiological dysfunction and psychic disturbances of the patients with penile torsion, a variety of surgery were used to eliminate its abnormality. A modificed of Nesbit procedure was used to correct the major malformation, several parallel ellipsoid patches which axis forms an angle of 45-degree with that of the penis were removed from the tunica albuginea on the convex side of the corpora cavernosa. Besides, stitches were necessary to suture the tunica albuginea to the opposite pubis. It was suitable plicate the tunica albuginea on the convex side of the corpora cavernosa for mild case. The penises of the patients had a normal appearance at both flexible and errect conditions during a 3-month follow-up. Penile torsion could be corrected satisfactorily by shortening the convex side of the corpora cavernosa with excision of ellipsoids or plication of the tunica albuginea, which angulates 45-degree with the penile axis. PMID- 10374542 TI - [Expression of p53 in nephroblastoma and neuroblastoma]. AB - We studed on the expression of p53 in 48 nephroblastomas and 32 neuroblastomas wit, immunohistochemical method of avidinbiotinperoxidase complex. 11 of the nephroblastomas were positive and all neuroblastomas were negative. Statistical analysis revealed that abnormal expression of p53 was significantly correlated with NWTS--II classification, NWTS--III clinical pathology stages and metastasis of nephroblastoma. The results indicate that abnormal expression of p53 is not only involved in the development, but also in the differentiation and metastasis of nephroblastoma. Therefore, detection of p53 expression may have significant clinical importance in the evaluation of prognosis of nephroblastoma. However, the absence of expression of p53 in neuroblastomas suggests that the inactivation of this gene does not play a significant role in the tumor's occurring and progression. PMID- 10374543 TI - [OKT3 treatment of refractory renal allograft rejection]. AB - Between January 1993 and June 1996, 44 episodes of refractory renal allograft rejection were treated with OKT3 to understand the curative effect of OKT3. Six of 7 accelerated and 31 of 37 acute rejection episodes were reversed. The reverse rate was 84.6%. Immediate response to OKT3 was in 25 and delayed response in 12 patients. The mean reverse time was 7 +/- 4 days in immediate response and 34 +/- 3 days in delayed response (P < 0.01). It was claimed that OKT3 had high effect in the treatment of refractory renal allograft rejection. Cytomegalovirus infection and cytokins release syndrome were the main side effect of treatment with OKT3. The delayed response would be associated with cytokins release syndrome and damage of rejection. Patients with lower titre of anti-OKT3 antibodies could be retreated with OKT3. PMID- 10374544 TI - [Foraminal and extraforaminal lumbar disc herniations]. AB - The severity of the clinical presentations of the foraminal or extraforaminal lumbar disc herniation (FLDH or EFLDH) and its response to conservative treatment were different. In order to make clear the reasons for that and then chose the best method of treatment, two types of FLDH or EFLDH were classified in a group of 23 cases according to the location of the protruded disc shown in the films of CT or MRI. Type I: the disc protruded up to the superior pedicle closely. Type II: the disc protruded laterally mainly with slight moving up. Then two subtypes were made, subtype a: symple FLDH or EFLDH: subtype b: if combined with posterior lateral disc herniation. In total of 23 cases there were 10 of type (I a: 8, I b: 2) and 13 of type II (II a: 7, II b: 6), among of which there were 14 FLDHs and 9 EFLDHs. The clinical presentations and the results of conservative treatment were compared between the two types. The results of statistical analysis showed that the severity of damage to the nerve root and its response to conservative treatment were mainly related to the location of the protruded disc to the superior pedicle rather than the protruded disc in the foramen or extraforamen. All of cases of type I had more severe symptoms and signs, poorer results of conservative treatment and were operated on finally. However some cases of type II only had slight or even no symptoms and signs at all, 50% of the patients in the type II were cured by nonsurgical methods. So it was suggested that the cases of type I be treated surgically mainly, whereas the type II initially be treated conservatively and if failed, go to operation. The approach lateral to the pars interarticularis for the discectomy had more advantages than the other methods and could be applied in most of cases. If combined posterior lateral disc herniation was the main pathologic change in the cases of type II laminectomy with partial facetectomy might be used. The classification is reliable as it is well consisten with the findings of operations. So it is valuable for the guidance of determining the method of nonsurgical or surgical treatment. PMID- 10374545 TI - [Clinical study of the relationship between the lateral recesses and the nerve roots]. AB - To explicate the relationship and the clinical signification between the normal or narrow lateral recesses and the nerve roots, we measured the diameter of the entrans zone of the lateral recess, the interval between the upper articular processes and the interval between the nerve root and ab line on 50 normal cases, 43 narrow cases and 32 stenosis cases with VIDS image analysis system. The results showed that the nerve root was in the center side of the ab line in the normal station, with the degrees of the degeneration and cohesion ncreasing, the nerve root was in the lateral recess side of the ab line, and was compressed by the lateral recess. The authors considered that the real clinical signification of the entrance zone of the lateral recess was danger to the nerve root, but the deciding factors were the degrees of the degeneration and cohesion of the upper articular processes. The pathological conditions that resulted in the stenosis of the lateral recess and dangered the nerve root such as disc, flavum ligament and posterior port of the fibra ring were discussed in the article. PMID- 10374546 TI - [Primary clinical experience of neuroendoscopy: report of 19 cases]. AB - 10 cases of basal ganglion and 6 of subcortical hematoma were evacuated. One case of temporal and 2 cases of para- and intra-ventricle arachnoid cyst were fenestrated towards the cistern or lateral ventricle. Burred holes were near the lesion, through which endoscope should run. We guided the endoscope to the target by stereotatic equipment in 10 cases of basal ganglion hematoma and 1 case of ventricle arachnoid cyst, and introduced endoscope by free-hand in other cases. Hematoma group: CT reexamination within 48 hours showed that the residual hematoma was less than 20%-30% in 9 of 11 cases. Cyst group: symptoms of all cases of arachnoid cysts were alleviated after intervention. CT after a month showed that fenestration entrance could be seen clearly in 2 cases. The cyst reduced 20%-30% in 1 case. There were no direct complications in the group. It was shown that operation by endoscope is minimal invasive and little complicated. PMID- 10374547 TI - [Cardiac operation via subaxillary and anterolateral subaxillary thoracotomy with cardiopulmonary bypass]. AB - From March 1995 to August 1996, 50 patients underwent cardiac operation through subaxillary and anterolateral subaxillary thoracotomy with cardiopulmonary bypass. 17 had atrial septal defect, 26 ventricular septal defect, 3 mitral stenosis and insufficiency, 1 isolated mitral insufficiency, 1 double chamber of right ventricle, 1 partial common atrioventricular canal, and 1 cor atriatriatum. All operations were successfully performed without technical difficulty. The cardiopulmonary bypass time was 40.19 +/- 17.17 minutes with aortic cross clamping time of 22.59 +/- 11.06 minutes. There were no operative death and complications. In conclusion, the subaxillary and anterolateral subaxillary thoracotomy incision appears to be a minimally invasive, safe, effective and cosmetic alterative to median sternotomy for cardiac operation. PMID- 10374548 TI - [Pedicled omentum graft for improvement of late-blood supply after myocardial revascularization with Nd:YAG laser in the treatment of ischemic heart disease]. AB - 26 dogs were devided into 3 groups. Selected left coronary artery branches of all dogs were ligated to make the models of myocardial infarction. The control group of 10 dogs received ligation of selected coronary artery branches(L group) only. Eight dogs were revascularized by Nd:YAG laser (LL group), and 8 dogs were revascularized and covered with pedicled omentum. All dogs were monitored by ECG, cardiac output, fractional 86Rb uptake and histological examination. The results showed that laser channels can perfuse ischemic myocardium and recover myocardial circulation to improve cardiac function immediately. However, the laser channels were closed after 4 weeks by clot, fibrosis and scarring which caused by thermal damage of Nd:YAG laser. The late result of laser myocardial revascularization with pedicled omentum graft is more better in perfusing myocardium and improving cardiac function than sole laser myocardial revascularization. PMID- 10374549 TI - [Thin split thickness skin grafting double taken from avulsed skin in treatment for skin avulsion in children]. AB - In order to utilize avulsed skin to cover skin defects, a new skin grafting technique, thin split thickness skin grafting double taken from avulsed skin, was used in 23 cases of severe skin avulsion. The skin grafts in profound layer were observed histologically. The results showed no difference between skin grafts in superficial layer and ones in profound layer, but the latter needed longer time to heal. Using this technique, we can obtain skin graft in double amount from avulsed skin. PMID- 10374550 TI - [Portal-systemic shunting and hemodynamics of prehepatic portal hypertensive rat models]. AB - Hemodynamics and relationship between portal-systemic shunting and portal pressure were measured by radioactive microsphere techniques in 31 rats after various degree portal vein constriction. Prominent hyperdynamic state was present in prehepatic portal hypertensive rats models. Both cardiac output and index were increased, the mean arterial pressure and pripheral resistance were reduced significantly. The total splanchnic blood flow was increased but splanchnic resistance reduced remarkably. Portal-systemic shunting flow was higher. There was a significant positive correlation between them, Y = 26.14 + 5.32X (r = 0.76, P < 0.001). For this reason, elevated systemic and portal venous flow provided main impetus for maintaining portal hypertension. PMID- 10374551 TI - [Measurement of urinary content of lactulose and mannitol by gas chromatography as an index of permeability of the gut]. AB - We established lactulose-mannitol(L-M) measurement method by gas chromatography and 9202 computer data processing system to test intestinal permeability. The urine output of L-M was in linear correlation to its sample concentration within working range. In an animal model of acute pancreatitis, lactulosesecretion increased in urine, together with increased L/M ratio. The measurement of lactulose-mannitol intestinal permeability by our method might serve as a predictor for early diagnosis of endogenous infection and sepsis. PMID- 10374552 TI - [Immunohistochemical change of endothelin-1 in acute lung injury and the role in its pathogenesis]. AB - On the rat acute lung injury model produced by intraperitoneal cavity injection of zymosan showed that the respiratory rate became slow at first time (1st day), subsequently became rapid. Pulmonary histology showed that there was an accumulation of neutrophils in pulmonary microvessels, neutrophils infiltration in interstitial tissue, interetital and olveolar edema, hyaline membrane, atelectasis. Positive expression of ET-1 in experimental rats was stronger than than contral rats. Image analysis showed that OD value was elevated, especially at the fist day. The quantity of ET-1 was corresponding to the degree of pathologic change of acute lung injury. It suggests that the ET-1 might play an important role in the mechanism of acute lung injury. PMID- 10374553 TI - [Methods to observe the experimental study of spinal cord injuries]. PMID- 10374555 TI - [Establishment and characterization of doxorubicin-resistant BGC-823/DOX of human gastric cancer cell line]. AB - To study drug resistance mechanism of gastrointestinal tumor, doxorubicin resistant cell line of human gastric cancer, we developed BGC-823/DOX, by increasing doses of doxorubicin. We completed chromosomal analysis, double time of cell grow, cell cycle distributions, drug resistance and cross-resistance, and studied reversing drug resistance. The results showed that chromosome of BGC 823/DOX was subtriploid karyotype. Cell morphology, double time, cell cycle distributions were different from its parent cells and 6.35 times more resistant to cytotoxic action of doxorubicin, cross-resistant to VP-16. Verapamil showed modified effect on doxorubicin resistance. BGC-823/DOX with doxorubicin resistance has many characteristics of gastrointestinal tumors. PMID- 10374554 TI - [Influence of preoperative on gastric cancer tissues and cells]. AB - We studied how gastric cancer tissues and cells changed as a result of preoperative chemotherapy. 82 patients with gastric cancer in TNM I-IV stage underwent preoperative selective vascular chemotherapy with routine quantity of FAM. Histological analysis of their postoperative specimens revealed the following results. (1) Cancer tissues necrosed in 56.1% patients. These necroses, due to the chemotherapy, were located around vascular vessels. (2) Cancer cells were found to change with karyopyknosis, karyorrhexis, coagulation and necrosis of cytoplasm. There were invasion of lymphocytes, inflammation edema, and fibroelastosis around cancer cells as well. Changes were also observed in vessels with proliferous intima and thrombus. Most of these changes were grade 2. This paper summarises that preoperative vascular chemotherapy can bring about evident histological and cell changes in different types of gastric cancer. This kind of chemotherapy is recommended as one of integrative treatments to gastric cancer. PMID- 10374556 TI - [The role of CDDP on cytotoxicity of TILs obtained from colorectal tumors]. AB - The effects of intravenous injection of cisplatin (CDDP) and the incubated with CDDP in vitro on the epitopes and cytotoxicity of TIL obtained from eight colorectal tumor patients were detected respectively by flow cytometer. There was a significant increase in CD3+/CD8+ subset in TIL in patients receiving intravenous injection of CDDP. And the cytotoxicity of these TILs increased significantly. The cytotoxities were positively correlated with the amount of CD3+/CD8+ subset in TIL. Raji cells incubated with CDDP in vitro showed increased susceptibility to TIL induced lysis. PMID- 10374557 TI - [Somatostatin and its secretory cells in tumor surrounding mucosa in colorectal cancer patients and its significance]. AB - The somatostatin (SS) levels in tumor surrounding mucosae were determined by RIA, and the SS secretory cells in these tissues were observed by immunocytochemistry in colorectal cancer patients. The mean SS level was higher in cancer-adjacent mucosa (CAM, 0-2 cm from the tumor) than in cancer-distant mucosa (CDM, about 5 cm from the tumor), and in CAM without atypical hyperplasia than in such mucosa with different grades of atypical hyperplasia (P < 0.01). In CAMs, there was a very significant negative correlation between the SS level and cell dysplasia (P < 0.01). The form and location of SS cells in all tumor surrounding mucosae were similar to the normal. Both the mean SS cell number and total positive degree were very significantly higher in CAM than CDM (P < 0.01). SS levels in CAMs were correlated positively with the SS cell numbers and the total positive degrees in CAM (P < 0.01). It is concluded that the elevation of SS level in CAM is mainly caused by the increase of SS cells, which secrete more SS. The change of SS in colorectal cancer surrounding mucosa may play an important role in inhibiting the development of CAM atypical hyperplasia or the tumor, and is a local defensive reaction in the body. PMID- 10374558 TI - [Gastric carcinoma in the young people]. AB - We studied the characteristics of the incidence, and prognosis of the gastric carcinoma in young people. The clinicopathological findings and follow-up data of the gastric carcinoma patients treated from 1976 to 1991 were compared in 624 olderly patients. The male to female ratio was 1:1.2 in young patients and it increased with age up to a peak: in the 50-59 year-old group (2.8:1). An increase of poorly differentiated tumors was found significantly in the young people (88.2%) compared with the olderly (P < 0.01). A slight preponderance of diseases in stage III and IV was found in the young people (72.5%, P > 0.1). There was no significant difference of resectability and postoperative actuarial survival between the two groups. Most young patients were diagnosed in the late stage. Early detection of the carcinoma in the young patients plays a significant role in improving their treatment results. PMID- 10374559 TI - [Endoscopic papillosphincterotomy for the treatment of pancreaticobiliary disease]. AB - From July 1987 to March 1996, endoscopic papillosphincterotomy (EST) was performed in 346 patients of 2,700 cases undergoing ERCP. Maj or indications for EST were choledocholithiasis, constrictive papillitis, chronic pancreatitis, biliary ascariasis and dysfunction of Oddi's sphincter. The EST was successful in 331 cases, ang the overall success rate was 95.7%. The stone discharge rate was 96%. The stricturotomy rate was 93.8%. The failure rate was 4.3% (15 cases). Clinical application and prevention of complications for EST were discussed. PMID- 10374560 TI - [Surgical treatment for advanced gallbladder cancer]. AB - To rise the resection rate and survival rate for advanced gallbladder cancer (GBC), we operated on forty cases of advanced GBC. 34 cases had obstractive jundice, and 8 palpable abdominal mass. Externded radical cholecystectomy was performed in 11 cases of advanced GBC in which the tumor invaded the surrounding organs or tissues but without distantce metastasis. Follow-up showed that the survival period was between 8 and 32 months (average 18.8 months), 1 and 2 years survival rates were 54.4% and 27.3%. Palliative operations were performed in other 29 advanced GBC cases of distant metastasis. All of the cases died within one year. We suggest that extended radical cholecystectomy is effective for advanced GBC. PMID- 10374561 TI - [37 patients with hepatic hydatid cyst rupturing into choledochus]. AB - 2785 patients with hepatic hydatid cyst were treated operatively since 1965 to 1995, and the hydatid cyst rupturing into choledachus occurred in 37 patients (1.3%). Clinical manifestations, diagnosis and operation were described. In epidemic area the possibility of hepatic hydatic cyst rupturing into choledochus should be considered when the patient suffers from stiffness of abdominal muscles, Jaundice, right epigastric pain, hepatic mass and positive reaction of Casoni test. Ultrasound and CT are major means for the diagnosis of this disease. Early operation is the treatment of choice. PMID- 10374562 TI - [Treatment of patients with dislocation of acromio-clavicular joint using kuntsher nail with steel wire: report of 27 cases]. AB - There has been much controversy about ways of operation of the traumatic dislocation of acromio-clavicular joint. Our studies revealed that internal fixation of acromio-clavicular joint using kuntsher nail with steel wire is superior to traditional internal fixation of crossing kuntsher nail. We treated 27 cases in this way who were followed-up for 22 months to 9 years. Excellent result was achieved in 25 cases (92.6%) and good in 2 (7.4%). No complication was observed. We conclude that the way of internal fixation using kuntsher nail with steel wire to treat the dislocation of acromio-clavicular joint has following advantages: (1) simple operation; (2) minimum injury to articular surface; (3) stable internal fixation; and (4) no side effect. PMID- 10374563 TI - [Correction of lower extremity deformity by unilateral axial dynamic fixator]. AB - Seventy-eight limb segments with various lower extremity deformity in 60 cases were treated by unilateral axial dynamic fixator (UADF). Bone healing of 77 osteotomy sites was primary union. One site was delayed union. The correction of limb axial line and recovery of joint function were excellent. The incidence of pintrack infection was 0.6%. The advantages of this device were minimum injury, simple procedures, stable fixation and reliable correction of deformity, promotion of callus formation and ossification by elastic fixation, less joint functional effect, and rare complication. The advandage of this method is that limb deformity can be adjusted to an ideal shape by using UADF. The average follow-up for 2.5 years indicated that application of UADF is an alternative method for the treatment of genu varus, genu valgus, and malunion of fracture. PMID- 10374564 TI - [A clinical review of 700 cases of coronary artery bypass grafting]. AB - To assess the changing trends in patients profiles, operative procedures, and the result of coronary artery bypass grafting (CABG) we reviewed the clinical data of 700 consecutive patients who had anisolated CABG (471, 67.3%), CABG combined with left ventricular aneurysm (170, 24.29%), CABG and valve procedures (48, 6.9%), CABG and ventricular septal defect repair (16, 2.3%) at the Fuwai Hospital. The patients were divided into group A (recent three years) and group B (before 1993). The incidence was significantly increased (P < 0.05) in diabetes, hypercholesterolemia, ventricular dysfunction, left main stem coronary artery disease and three vessles lesion. There was a high incidence of hypertension (41.3%), old myocardiac infarction (36.4%), combined left ventricular aneurysm (24.29%) and IABP needed (9.4%), however no significantl difference was notes between the two groups. Internal mammary artery was used in group A (25.6%) and group B (2.6%) (P < 0.005). The hospital mortality (group A, 2.7% and group B, 9.6%) and perioperative myocarial infarction (group A, 3.2% and group B, 9.0%) decreased significantly (P < 0.005). We conclude that despite a high incidence of hypertension, diabetes, hypercholoesterolemia, old myocardial infarction, ventricular dysfunction and diffuse coronary artery disease the patients tolerate CABG surgery well and obtain a good result. PMID- 10374566 TI - [Clinical application of a modified small posterolateral thoracotomy in general thoracic surgery]. AB - To assess the application of a modified thoracotomy approach in general thoracic operation. 311 general thoracic operations were performed using a modified small posterolateral thoracotomy surgical procedures included lobectomy plus lymph nodes dissection in 186, pneumonectomy in 23, esophagectomy for esophageal cancer in 23. The modified method of thoracotomy had following advantages: (1) reduce postoperative morbidity; (2) reduce bleeding and bank blood transfusion; (3) shorter operating time. This study suggested that the modified small posterolateral thoracotomy have some advantages than standard approach. It should be used in some selected thoracic operations. PMID- 10374565 TI - [Surgical treatment of thoracic aortic aneurysm]. AB - In 80 patients with thoracic aortic aneurysm were treated in our hospital, 62 were males and 18 females. Their age ranged from 20 to 73 years. The aneurysm lesions involved ascending aorta in 45 patients (Marfan's syndrome), aortic isthmus segment in 3, descending aortic aneurysm in 26, and thoraco-abdominal aneurysm in 3. Thoracic aortic aneurysm with dissecting aneurysm were found in 49. Stanford type A was present in 28 cases patients and type B in 21.61 patients underwent surgery including Bentall's operation in 35, Wheat's operation in 3, resection of discending aortic aneurysm with vascular prosthesis grafting in 14, and other types of operation in 9. Operative mortality rate was 6%. In the medically treatment group of patients, spontaneous rupture of the aneurysm occurred in 8. Among them, 5 died in hospital and another 2 at home. In this series of patients, Marfan's syndrome and dissecting aneurysm were the predominant pathological lesion. Since acute rupture of dissecting aneurysm is the main cause of death, surgical treatment should be immediate after the diagnosis is established. In the chronic dissecting aneurysm, early surgical treatment also should be considered. PMID- 10374567 TI - [Primary renal epithelioid angiosarcoma with transitional cell carcinoma in renal pelvis]. AB - To understand the clinical characteristics of primary renal epithelioid angiosarcoma. We used clinical pathology, immunohistochemistry and electron microscopy. Analysis and discussion about the case were combined with a review of the literature. We first report a female case of the left renal epithelioid angiosarcoma and concomitantly, transitional cell carcinoma developing from the renal pelvis at age 69. Epithelioid angiosarcoma showed positive results for FVIIIRA+, UEA, CD31, Cytokeratin and EMA. Weibel-Palade bodies were identified by electron microscopy. Primary renal epithelioid angiosarcoma was a rare clinical entity with a poor prognosis, which mimics epithelial tumor, both morphologically and immunohistochemically, and may be lead to misdiagnosis. The combined use of endothelial cell markers including FVIIIRA, CD31, UEA and electronmicroscopy can confirm the diagnosis of this neoplasm. PMID- 10374568 TI - [Extracorporeal shock wave lithotripsy for uric acid stones in upper urinary tract]. AB - Since 1987, 121 patients with uric acid stones in upper urinary tract have been treated by an EDAP LT-01 ultrasound-guided piezoelectric lithotriptor. One hundred and twenty-one patients had 167 renal stones 0.5-3.5 cm in diameter (mean 0.86 cm), 9 had ureteral stones 0.6-1.3 cm (mean 0.84 cm). Auxiliary treatment were given to 2 patients with upper ureteral stones, the others were treated by ESWL in situ. A success rate of 80.99% (98 patients) was achieved in one session, 12.40% (15) in two sessions, 4.96% (6) in three sessions, and 1.65% (2) in four sessions. No serious complications were encounterd. Therefore, ESWL with the ultrasound-guided piezoelectric lithotriptor is considered to be a satisfactory procedure for the treatment of uric acid stones. The ultrasound localization of uric acid stones, the shocking methods during ESWL and the application of alkaline medicine after ESWL were discussed. PMID- 10374569 TI - [Perioperative plasma amino acid spectrum analysis in portal hypertensive cirrhotic patients undergoing portacaval H-graft shunt]. AB - Perioperative plasma amino acid spectrum (PAAS) was analysed in 10 portal hypertensive patients (PH) receiving portacaval H-graft shunt (D = 8 mm) and 10 gastroenteropathic patients (GE) without hepatic disease. In both groups, arterial, peripheral and hepatic venous blood was drawn for analysis immediately pre- and post-operation on the table, postoperative day 1 and 3, respectively. It was found that amino acid concentration decreased in both groups but in PH group it was changed significantly. The preoperative branched chain amino acid (BCAA)/aromatic amino acid (AAA) ratio was less than 2 in the PH group and larger than 3 in the GE group after operation. The ratio did not change significantly in both groups after operation. In the PH group, the pre- and post-operative BCAA and AAA level in hepatic vein was higher than in other sites. The immediately BCAA and AAA levels were lower than preoperative levels. Among the changes the postoperative decrease of VAL and LEU was significant. Both BCAA and AAA reached preoperative level at postoperative day3. The results reveal that small diameter portacaval shunt has no significant influence on PAAS. The temporary decrease of amino acid levels postoperatively may be related to the stress of operation and liver impairement. PMID- 10374570 TI - [The effect of blood-pancreatic juice barrier on antitumor drugs excretion]. AB - Chemotherapy is one of important adjuvant therapies for patients with pancreatic adenocarcinoma, but the mechamism 5-FU and MMC distributed over the pancreas is not known at home and abroad. Based on determining the ratio of the concentration of several kinds of antibioticsin plasma and pancreatic juice, we used fourteen dogs to study the distribution and the relationship of agents in plasma and pancreatic juice after giving 5-FU and MMC intravenously. At the same time, we observed some patients who received Whipple's operation. The results showed that 5-FU and MMC in plasma can penetrate the pancreatic tissue and cross the blood pancreatic juice barrier (BPJB). Both agents suitable for the adjuvant chemotherapy of pancreatic adenocarcinoma. PMID- 10374571 TI - [c-met expression in human hepatocellular cancer]. AB - Using Northern blot method, we examined the expression of c-met in human hepatocellular cancer. Of 30 patients tested, the expression of c-met mRNA was not significantly different in HCC tissues from corresponding noncancerous parenchyma (P > 0.05). There was no significant relationship between c-met expression and grade of differentiation, stage of clinic, AFP, HBsAg (P > 0.05). Our results suggest that c-met may play different roles in the pathogenesis and metastasis of human hepatocellular carcinoma. PMID- 10374573 TI - [Experimental study of vascular gene transfer using soluble stent]. AB - We assessed the possibility of gene transfer into anastomotic arteries in vivo using soluble stent containing Adv5-CMV/LacZ. After being soaked in a high concentration solution of glucose containing Adv5-CMV (control group) or Adv5 CMV/LacZ (treatment group) for 30 minutes, we inserted soluble stents into the lumina of cut rat carotid arteries, and end-to-end anastomoses of cut rat carotid were performed with standard microvascular surgical technique. 16 rats were killed after two weeks, the segments of anastomotic carotid arteries were prepared for assessing beta-galactosidase activity and histochemical staining. In the control group, the anastomotic arteries did not have detectable level of beta galactosidase expression. In the treatment group, the amount of beta galactosidase expression was 9.80 x 10-3 u/g tissue. Microscopic examination of histochemically stained arteries demonstrated that gene transfered not only to endothelial cells but also to smooth muscle cells, and all anastomotic arteries were transfered in the treatment group, but none of arteries revealed blue in the control group. The results of this experimental study suggested that soluble stent be a new method of direct gene transfer into arteries in vivo. PMID- 10374572 TI - [Revascularization of isolated bone in dogs]. AB - To elucidate whether the revascularization of isolated bone could be built and what is the influence on osteogenesis. We studied eighteen adult mongrel dogs and divided them into three groups in random. Osteotomy was performed on the two condyles of the tibia and the periosteum was removed. Two pieces of silica gel sheet were inserted into the gaps of osteotomy to block blood supply, then immobolization was made. The vascular bundle of anterior tibia artery and vein was separated and ligated. Subsequently, it was directly implanted into the isolated bar of tibia (Group A, n = 7). It implanted after holes were made (Group B, n = 7). Controls (Group C, n = 4) were observed without transplantation. The animals were investigated by angiography, casting mode of vascular net, mass and histologic and electromircroscopeic examination at different intervals. Osteonecrosis was occurred in the tested bone because of ischemia. The proliferation of vascular net was occurred and prosperous osteogenesis was found in the medullary cavity after transplanting a vascular bundle. Revascularization of necrotic bone was able to be established by transplanting a vascular bundle. Both membranous and lndochondral ossification play an important role in osteogenesis. PMID- 10374575 TI - [Fracture repair and cytokine regulation]. PMID- 10374574 TI - [Detection of mRNA expression of endothelin-1 in organs on severe burn rat model with in situ hybridization technique]. AB - After the dynamic observation of postburn endothelin changes in plasma on severe burn rat model, we detected ET-1 mRNA expression and cell localization in some organs such as heart, lung, liver and kidney with in situ hybridization technique. We that endothelin concentration in plasma rapidly increased postburn (it reached its climax at 6th hour and maintained fairly high level even at 24th hour postburn). Meanwhile in situ hybridization indicated that ET-1 mRNA expression intensity and number of positive cells rapidly increased postburn. The expression climaxes of which appeared decreasing in turn from kidney, liver, lung to heart and the types of positive cells increased. This may due to the endothelin's functions of contracting blood vessels and regulating the redistribution of blood flow and be helpful for guaranteeing blood perfusion of important organs under shock state, but the overexpression of ET-1 mRNA may make the ischemic and anoxic damages to organs more severe. PMID- 10374576 TI - [The International Program of Global Elimination of Silicosis]. PMID- 10374577 TI - [Studies on cytotoxicity of man-made mineral fiber to alveolar macrophage]. AB - OBJECTIVE: To compare the difference in cytotoxicity to human alveolar macrophage caused by ten kinds of man-made mineral fiber. METHODS: Ten kinds of man-made mineral fiber prepared by Japan Fibrous Materials Research Association (JFM) were used with alveolar macrophage extracted from guinea pigs in vitro as the target cells. Changes in the production of O2- and H2O2, levels of glutathione and cellular free calcium ion (Ca2+) in alveolar macrophage were determined. RESULTS: Most of the 10 kinds of JFM fiber could cause increased production of O2- and H2O2, reduction of levels of glutathione and increase of cellular free calcium ion (Ca2+) in alveolar macrophage, to a extent less than that caused by chrysotile. CONCLUSION: In general, the cytotoxicity of JFM fiber was less than that of chrysotile. Although these mineral fiber could be used as substitutes for asbestos, they still could cause certain harm to human bodies. PMID- 10374578 TI - [Comparison of changes in oncogene of human embryo lung cells before and after chemical modification for chrysotile]. AB - OBJECTIVE: To seek for a suitable chamical agent for surface modification of chrysotile. METHODS: The effects of aluminum citrate, mixed rare earth and sodium selenite on c-Ha-ras oncogene and P21ras protein in the transformation of human embryo lung cells caused by chrysotile were studied. RESULTS: The transcription level of c-Ha-ras oncogene and the content of P21ras in those exposed to chrysotile were significantly higher than those in the controls. The transcription level of c-Ha-ras oncogene and the content of P21ras in those exposed to chrysotile treated in advance with three kinds of chemicals mentioned above were significantly lower than those in untreated. CONCLUSION: Treatment of chrysotile with three kinds of above-mentioned chemicals in advance could reduce its carcinogenecity in human beings. PMID- 10374579 TI - [Changes in blood levels of nitric oxide, oxidation and lipoperoxidation in patients with silicosis]. AB - OBJECTIVE: To explore the relationship between nitric oxide, oxidation, lipoperoxidation and silicosis. METHODS: Blood plasma levels of nitric oxide (P NO), vitamin C (P-VC), vitamin E (P-VE), beta-carotene (P-beta-CAR), lipoperoxidase (P-LPO), and erythrocyte activities of superoxide dismutase (E SOD), catalase (E-CAT), glutathione peroxidase (E-GSH-Px) and lipoperoxidase (E LPO) were measured in 73 patients with silicosis and 60 healthy subjects. RESULTS: The average levels of P-VC, P-VE, P-beta-CAR, E-SOD, E-CAT and E-GSH-Px were significantly lower and the average levels of P-NO, P-LPO and E-LPO significantly increased in the silicosis with varied severity of the disease and lung function status more than in the controls. There was linear relationship between all the above indices and length of the disease. Stepwise regression analysis showed that the severity of the disease and lung function status correlated most closely with the levels of P-NO, P-VE, E-SOD and E-LPO. CONCLUSION: Metabolism of nitric oxide was abnormal, balance between oxidation and antioxidation was disturbed seriously, and oxidation and lipoperoxidation reaction pathologically exacerbated in the bodies of the patient with silicosis. PMID- 10374580 TI - [Studies on effects of cytokine released from alveolar macrophage induced by mineral dust on lung fibroblast]. AB - OBJECTIVE: To understand the role of cytokine released from alveolar macrophage (AM) in lung fibrosis caused by mineral dust. METHODS: Rabbit's AM obtained by lavage was cultured with mineral dust in vitro. Activities of tumor necrosis factor (TNF) and interleukin 6 (IL-6) in its supernatant were determined with isotope labelling method and MTT colorimetry, respectively. Human fetal lung fibroblast WI-138 was cultured with this supernatant. Proliferation of fibroblast and synthesis of collagen were examined by 3H-thymidine (3H-TdR) and 14C-proline (14C-Pro) incorporation and its total hydroxyproline (HOP) level was analyzed by chloramine-T method. RESULTS: Proliferation of lung fibroblast and synthesis of collagen could be enhanced by the supernatant containing AM induced by quartz, asbestos and uranium mineral dust, with 3H-TdR incorporated counts per minute (cpm) of 6,584, 3,848 and 6,893 in the group of 100 micrograms 3H-TdR and 14C-Pro incorporated 27,952, 13,416 and 18,538 in the group of 200 micrograms respectively, which were significantly higher than those in the control group. Total HOP levels in the culture media for lung fibroblast enhanced by AM supernatant were 22.41, 24.00 and 21.39 micrograms/ml, respectively, and was significantly higher than that in the control group (12.91 micrograms/ml). Release of TNF and IL-6 could be stimulated by mineral dust, such as quartz, asbestos and uranium mineral dust, and their activities were significantly higher than those in the control group, with those of 1,396, 1,198 and 852 U/ml in TNF group and 1,336, 1511 and 1,335 U/ml in IL-6 group, respectively. Proliferation of lung fibroblast and synthesis of collagen could be inhibited by antibody against TNF and interferon-r (gamma), and the effect of the latter was weaker than that of the former on inhibition of fibroblast proliferation and the effect on collagen synthesis was just in the opposite direction. CONCLUSION: Lung fibrosis caused by mineral dust correlated with abnormal expression of TNF and IL 6. Antibody against TNF and gamma interferon could antagonize the effect of NTF and IL-6. PMID- 10374581 TI - [Dynamic analysis of incidence of silicosis in uranium miners]. AB - OBJECTIVE: To understand the laws and characteristics of incidence of silicosis and to evaluate the effectiveness of prevention and treatment measures for it in uranium miners. METHODS: An epidemiological study was carried out in three uranium mines and two geological prospecting teams for uranium mines. RESULTS: Exposure to dust in all patients with silicosis initiated in 1950s to 1960s. Dust concentration and detection rate of silicosis have been decreasing, the length of employment in uranium mines and age at occurrence of the disease, interval between lower and upper stages of the disease, its duration and age at their death all have showed an increasing trend since the founding of the mines and geological prospecting teams more than thirty years ago. Those with delayed occurrence of the disease accounting for 20.31% of the total cases. The length of employment in the cases exposed to uranium dust averaged 7.47 years, and the age at occurrence averaged 36.74 years, less than in those exposed to mixed non uranium dust. CONCLUSION: Effectiveness of comprehensive prevention and treatment measures for silicosis in uranium mines is significant. The average length of employment and age of the occurrence of the silicosis in those exposed only to uranium dust showed and younger. PMID- 10374582 TI - [An in vitro study on toxic effect of vanadium-titanium-magnetite dust on alveolar macrophage in rabbits]. AB - OBJECTIVE: To study the toxic effect of vanadium-titanium-magnetite (VTM) dust on alveolar macrophage (AM) and its hazardous extent. METHODS: Survival rates, morphology and function of AM were compared in rabbits exposed to dust of VTM, vanadium oxide, titanium dioxide and silica in various doses and length of time with in vitro cell culture and putamen membrane cover glass transmission electron microscopy, and changes in activities of lactic dehydrogenase (LDH) and acid phosphatase (ACP) in cell culture were measured. RESULTS: Exposure to all the four kinds of dust could lead to decrease in survival rate of AM, increase in activities of LDH and ACP in the cell culture, and changes in their morphology and function to the extent dependent on the nature of dust. CONCLUSION: Toxic effect of exposure to VTM dust was lower than that to vanadium oxide and silica, but higher than that to titanium dioxide, which had slight toxic effect. PMID- 10374583 TI - [Analysis of HLA-DRB1 allele in patients with chronic bronchitis]. AB - OBJECTIVE: To study the difference in HLA-DRB1 allele frequencies between patients with simple chronic bronchitis and with coal worker's pneumoconiosis (CWP) complicated with chronic bronchitis. METHODS: HLA-DRB1 allele frequencies were measured in 37 patients with simple chronic bronchitis and 33 with CWP complicated with chronic bronchitis of Han ethnicity in Shanxi Province using polymerase chain reaction with sequence specific primer, in comparison with those in normal population of the same province. RESULTS: Allele frequency of HLA-DRB1 * 120X was 24.32% in those with simple chronic bronchitis, and that of HLA-DRB1 * 040X was 22.73% in those with CWP complicated with chronic bronchitis, significantly increased than that in normal people with both P < 0.01. There was no significant difference in other alleles between the three groups. CONCLUSION: It suggests that in Han ethnicity of Shanxi Province HLA-DRB1 * 120X allele frequency is associated with simple chronic bronchitis and that of HLA-DRB1 * 040X is associated with CWP complicated with chronic bronchitis. PMID- 10374584 TI - [Studies on incidence of malignant tumor in workers exposed to dust in a mine in 30 years]. AB - OBJECTIVE: To investigate incidence of malignant tumor in workers exposed to dust in a mine during the past 30 years. METHODS: A retrospective cohort study was conducted in 16,711 workers exposed to dust and 7,598 non-exposed workers. RESULTS: Incidence of lung cancer in exposed workers ranked the first place, whth an SMR of 2.648, as compared with that of non-exposed workers. Incidence of lung cancer in the dust-exposed workers with a longer duration of employment was significantly higher than in those with a shorter one. Incidence of lung cancer in exposed workers with a wetoperation mode was lower than that in those with dry operation mode. CONCLUSION: Malignant tumor, especially lung cancer, occurred more frequently in the workers exposed to dust, which could be a potential risk factor contributing to carcinogenesis. PMID- 10374585 TI - [Molecular epidemiology of TT virus infection in some parts of China]. AB - OBJECTIVE: To study the molecular epidemiology of new hepatitis TT virus (TTV) infection, its distribution in population and its role in pathogenesis of hepatitis in some parts of our country. METHODS: TTV DNA in serum samples was detected by nested-polymerase chain reaction, partial gene of various geographic strains of TTV was cloned and sequenced, and their genetic variation was analyzed. RESULTS: Forty-four of 112 cases with non-A to G hepatitis from Shenzhen, Guangdong province, Nanjing, Jiangsu province, Beijing and Shengyang, Liaoning province were positive for TTV DNA with a positive rate of 42.9%, but only three of 102 cases of hepatitis A-G was positive, with a positive rate of 2.9% (chi 2 = 42.8, P < 0.01). Positive rate of TTV DNA was significantly higher in blood donors with abnormal ALT but without infection markers of hepatitis A-G (34.6%) than in those with normal ALT (16.8%) (chi 2 = 4.5, P < 0.05). Sequencing analysis showed that more than 98% of their nucleotides were analogous between strains of TTVCHN002 from Shenzhen and Nanjing, and TTVSHB015 from Beijing, and more than 97% analogous between the above-mentioned strains and Japanese ones. CONCLUSION: There existed TTV infection in both north and south China. TTV infection correlated closely with abnormal ALT, which might be an important pathogen for non-A, non-B, hepatitis G. There were somebodies infected with TTV in the normal healthy population, similar to that of "chronic carrier status" in hepatitis B. PMID- 10374586 TI - [Changes in serum levels of total amino acids in young children with congenital heart diseases]. AB - OBJECTIVE: To study the implication of early surgical operation on the improvement of protein malnutrition in young children with congenital heart diseases. METHODS: Sixteen kinds of serum free amino acids were determined with an automatic analyzer for 72 cases with congenital heart diseases before and after surgical operation and 51 healthy children. RESULTS: Serum total amount of essential amino acids reduced in children with congenital heart diseases, mainly of branch-chained amino acids, especially significant in young ones. Serum amino acids restored to normal level one month after surgical operation. CONCLUSION: There existed protein malnutrition in children with congenital heart diseases, especially in young ones. Surgical operation for them in a suitable time will improve their protein nutritional status. PMID- 10374587 TI - [Effect of various levels of vitamin K intake on bone metabolism of rat]. AB - OBJECTIVE: To explore the effect of various levels of vitamin K intake on bone development. METHODS: Forty weanling Wistar male rats were divided into four groups. In one group, 1% sulfadiazine was added to regular diet (vitamin K 50 micrograms/kg) to induce vitamin K deficiency. In the other three groups, the vitamin K levels in diets were 50 micrograms/kg, 300 micrograms/kg, 2,550 micrograms/kg respectively. Twelve weeks later, the rats were killed and the effects of the different levels of vitamin K intake on bone development were evaluated by the parameters of calcium metabolism, bone metabolic biochemistry, and bone mineral density (BMD). RESULTS: Vitamin K did not affect the intestinal absorption of calcium. Vitamin K deficiency led to the high levels of urinary calcium and hydroxyapatite excretion, suggesting an increase of bone absorption. Different levels of dietary vitamin K significantly affect circulating osteocalcin and OCbound content. The level of serum2 osteocalcin, OCbound and BMD elevated with the increase of dietary VK levels, whereas the parameters of PTH (thrombo plastin time) were not different between all groups. CONCLUSION: Vitamin K can enhance the bone development of rat. The rat vitamin K requirement may be higher than that of the current recommendation (50 micrograms/kg). PMID- 10374588 TI - [Effects of perinatal care on low birth weight]. AB - OBJECTIVE: To explore the relationship between low birth weight and maternal factors during perinatal period. METHODS: Data of perinatal care were collected, sorted out and analyzed for 9,337 live births during 1993 to 1996, including 269 with low birth weight, in Xiucheng District of Jiaxing City, Zhejiang Province. RESULTS: Incidence of infants with low birth weight (less than 2,500 grams) averaged 2.88%, and appeared a gradual decreasing trend during the four years. Low birth weight correlated with maternal education, body height, number of gestations, antenatal check-ups, gestational age and the sex of babies. CONCLUSION: Systematic management of perinatal care for pregnant women will benefit to lowering the incidence of low birth weight, and perinatal and infant mortalies. PMID- 10374589 TI - [Studies on stroke and blood lipid level]. AB - OBJECTIVE: To elucidate the relationship between blood lipid levels and cerebrovascular diseases. METHODS: Seven indicators for blood lipid were determined in 208 patients with cerebrovascular diseases. Triglyceride (TG), total cholesterol (TC) and high-density lipoprotein-cholesterol (HDL-C) were measured with enzyme methods, apolipoprotein A-I, apolipoprotein B-100 (Apo-B100) and lipoprotein (alpha) [Lp(alpha)] were measured with immune multifocal method, and low-density lipoprotein-cholesterol (LDL-C) was calculated according to Friedewald formula. RESULTS: Levels of TG, TC, LDL-C, Apo-B100 and Lp(alpha) in patients with cerebral infarction were significantly higher than those in the controls, and levels of TG and Apo-B100 in them were also significantly higher than those in the patients with cerebral hemorrhage. There was no significant difference in blood lipid levels between the patients with cerebral hemorrhage and the controls. CONCLUSION: It suggests that disturbance of blood lipid metabolism is a risk factor for cerebral infarction. PMID- 10374590 TI - [Dynamic variation of incidence of neural tube defects during 1988 to 1992 in China]. AB - OBJECTIVE: To study the variation trend in incidence of neural tube defects (NTDs) during 1988 to 1992 in China. METHODS: Data were collected by hospital based monitoring, with women of gestation of 28 weeks to seven days after birth as study subjects. And, 3,246,408 perinates were monitored in the National Birth Defect Monitoring Network in 30 provinces, municipalities and autonomous regions all over the country, and 7,778 cases of NTDs during 1988 to 1992 were recorded and analyzed. RESULTS: Incidence of NTDs was 18.0 per ten thousand in 1992, 28.9% reduction from 25.3 per ten thousand in 1988, and yearly average reduction of 8.2%. Incidence of NTDs decreased by 30.5% and 34.2% in rural and urban areas, respectively, by 30.5% and 27.8% in the north and the south, respectively, and incidence of anencephaly and encephalocele fell by 34.2% and 48.5%, respectively. CONCLUSION: Incidence of NTDs showed a falling trend in China, more decrease in rural area and in the north than in urban areas and in the south, respectively, and incidence of anencephaly and encephalocela fell faster. PMID- 10374591 TI - [Uses of nasal lavage in occupational medicine and environmental medicine]. PMID- 10374592 TI - [Risk factors of epidemic spread of HIV infection in China]. PMID- 10374593 TI - [Long-term result of free forearm skin flap for repair of soft tissue defects of the oral and maxillofacial regions]. AB - To evaluate the long-term result of free forearm skin flap in the repair of soft tissue defects of the oral and maxillofacial regions, 26 cases which had received radical resection of maxillofacial tumors were follow-up for 4.5 years. Twenty cases, having complete data were analyzed. In this series, There were 8 males and 12 females, with ages ranged from 40 to 69 years old. The size of the flaps ranged from 4 cm x 5 cm-6 cm x 13 cm. The radial artery and the cephalic vein were used as the donor vessels, and the maxillary artery, superior thyroid artery, external jugular vein and the anterior jugular vein were prepared as the recipient vessels. According to the shape, colour, temperature, sensation, mucosoid degree of the flap, the blood supply and function of hand and the configuration of the forearm, the overall results of the recepient regions in 20 cases were all satisfactory and the overall results of 16 cases donor regions were satifactory in 16 cases. The results were poor in 4 cases. The conclusion were: 1. Free forearm skin flap was worth trying in the repair of soft tissue defects of oral region; 2. The radial artery need not to be reconstructed because of the abandant vascular net-work in the upper limb and 3. The residual scar on the forearm was the main shortcoming, but most of the patients could tolerate it because of the obvious advantages received from the operation. PMID- 10374594 TI - [Reconstruction of the floor of mouth with facial artery musculocutaneous flap]. AB - In order to study the clinical efficacy of facial artery musculocutaneous flap on repairing the defect of the floor of mouth, 21 patients had received this type of treatment from 1991 to 1997. The size of the flaps ranged from 8.0 x 3.4 cm to 12.1 x 5.4 cm and the average age of these patients was 59.5 years old. The donor site was closed directly. Nineteen flaps survived completely, while necrosis occurred at the apex of the other 2 flaps, which healed by ordinary management. The applied anatomy of the flap and the design and the main points of the operation were reported in details. The advantage of the flap and the prevention of facial malformation following operation were discussed. The conclusion was that this type of flap was ideal for reconstruction of the defects of floor of the mouth. PMID- 10374595 TI - [Application of MEDPOR surgical implant in the craniofacial reconstruction]. AB - Since November 1996, 20 cases of craniofacial deformities, either from congenital or traumatic, were treated with MEDPOR surgical implant made from a linear high density polythylene. The animal experiment had shown that the MEDPOR had good organotrophic characteristics allowing tissue ingrowth. The biocompatibility studies in vitro and in vivo had shown that the MEDPOR biomaterial was free from any observable systemic or cytotoxic effect. In the clinical application, it was found that the MEDPOR could be easily modeled and maintained. Because of the ability to induce tissuee ingrowth, the tenacity and stability of the material were enhanced. A total of seven cases of cranial defects, 8 cases of periobital defects or depressed periobital regions, 2 cases of traumatic auricular defects, 2 cases of traumatic saddle nose and 1 case of maldevelopment of mandible angle were treated. All of the cases were followed up for 6 months, the results were satisfactory. PMID- 10374596 TI - [Histological evaluation of collagen-hydroxyapatite composite as osseous implants in the repair of mandibular defect]. AB - To observe the collagen-hydroxylaptite composite in the repair of bone defect, ten minipigs were chosen to make a mandibular dafect measuring 2 cm in diameter and the composite was implanted, while the use of autogenous bone graft and the blank wese served as control. On the 4, 8, 12, 24 and 48 weeks after the operation, the animals were sacrificed and the samples were examined under light microscope. The result showed that: no infection or necrosis occurred. The composite coalesced with host bone and the outcome was similar to that of the autogenous bone graft. No foreign body giant cells or vacuum left from osteonecrosis was observed. It was suggested that the composite had the advantage of abundant supply, easy to handle and no harm. The biocompatibility was good and might be hopeful as a bone substitute. PMID- 10374597 TI - [The characteristics of plasma--sprayed coatings of hydroxy apatite in vitro]. AB - In order to investigate the stability of Hydroxyapatite (HA) coated material, the plasma-sprayed coatings of HA were divided into four groups: 1. Keeping in water vapour at 125 degrees C, with a pressure of 0.15 MPa for 6 hr; 2. Heating at 650 degrees C in air for half an hr; 3. Keeping in water vapour at 490 degrees C, with a pressure of 0.01 MPa for 2 hr; and 4. The control. The XRD, FTIR analysis and the dissolution test were carried out. The results showed: 1. The degree of crystalization in XRD analysis was 3. > 1. > 2. > 4.; 2. The ampitude of OH- peak in FTIR analysis was 3. > 1. > 2. > 4. and 3. The dissolution rate in tris-HCl buffer was 3. < 1. < 2. < 4. The conclusions were 1. The treating with water vapour could decrease the transformation temperature which was needed to convert the amorphous phase into cystalline phase; 2. Water vapour treatment could accelerate the transformation of Z-TCP, TCPM into crystalline HA; 3. Water vapour treatment could promote the structural integrity of plasma--sprayed coated of and HA and 4. Water vapour treatment could lower the dissolution rate of HA coated in Tris-HCl buffer. PMID- 10374598 TI - [Primary microsurgical repair of multi-structural defects of hand]. AB - Forty-eight cases of multi-structural defects of hands were primarily repaired or reconstructed from July 1989 to 1997. The structural defects included: the defects of radial or ulnar aspect of hands involving fingers and skin, multiple fingers defects and the fingers and skin defects of whole hand. In this series, there were 32 males and 16 females with age ranged from 17 to 46 years old. The composite tissue grafts were obtained from wrap-around flap or 2nd toe skin flap of the foot. The result showed that composite 108 tissues transplantations, or 48 cases, were all survived. After a follow-up of 38.5 months (ranged from 5 months to 6 years), the grasp, pinch and opposition function of the reconstructed finger were restored, the two-point discrimination sensation was 4 mm-12 mm. Most of the patients had resumed their original works. So that the primary repair of multi structural defects of hands by composite tissues transplantation was feasible and valuable, but thorough debridement and skilled microsurgecal technique were required. PMID- 10374599 TI - [The treatment of fracture of patella by internal fixation with tension band from musculofascial tendon]. AB - In the study of the efficacy of internal fixation with tension band of musculofascial tendon in the treatment of fracture of patella, 52 cases were reported. After a following-up of 6 months to 13 months the bone healing was observed, in 7.5 weeks in average, and the function of the knee joint had recovered to normal or almost normal. It was concluded that the treatment of fracture of patella by internal fixation with tension band from musculofascial tendon was a ideal and practicable method. PMID- 10374600 TI - [Clinical applyeation of neural stump buried into muscle for the prevention and treatment of neuroma]. AB - In order to verify the effectiveness of neural stump buried into the muscle in the prevention and treatment of neuroma, 17 cases were reported, in which 8 cases having 19 painful neuromas and 9 cases having 13 amputated meural stumps, buried into muscle. They wese followed up for 6 months to 40 months, It was shown that good and excellent results were obtained and no evidence of neuroma was observed in all cases except in one which had painful neuroma occurred from the failure of embedment of the neural stump into the muscle. The conclusion was that the neural stump buried into muscle was an effective method for the prevention and treatment of neuroma. PMID- 10374601 TI - [Repair of massive bone defect with free vascularized fibular graft]. AB - In the study of repair of massive bone defect with free vascularized fibula graft, 13 cases were reported, in which traumatic defect in 7 cases, segmental resection of bone from tumors in 5 cases and osteomylitis in 1 cases. They all were treated successfully with vascularized fibular graft. After a follow-up of 6 months to 7 year, bone healing was observed with satisfactory and rehabilitation of functions. In one case, fatigued fracture occured twice due to early walking. It was concluded that free vascularized fibular graft was very helpful in the repair of massive bone defect, but prolonged external fixation after operation might be important to prevent fractur of grafted bone. PMID- 10374602 TI - [Experimental study of the effect on growth of Schwann cell from chitin and chitosan in vitro]. AB - In order to study the effect of chitin and chitosan on the growth of Schwann cell (SC) of rats in vitro, the SC was isolated from sciatic nerve and brachial plexus of new-born rats. After the enzymatic and mechanical dissociation, the cell suspension was vaccinated on chitin membrane and chitosan fluid-coated glass coverslips. Then, the growth of SC was examined at 1, 3, 7 days after culture under light microscope and scanning electron microscope. The results showed that 94 percent of the cell grown from was SC and only 6% was fibroblast (FB), while that of the control SC 71% and FB 29% in population. The number of SC in chitosan suspension was more than that in chitin. Therefore, the conclusion was that the chitin and chitosan was histocompatible to SC, and chitosan suspension was superior to chitin, and both could inhibit the growth of fibroblast. PMID- 10374603 TI - [Processing of ceramiclike xenogeneic bone and experimental study of its bone formation from composite graft combined with bone marrow]. AB - Ceramiclike xenogeneic bone (CXB) was obtained from the fresh bone of pig ribs being treated by physical and chemical methods to deprive of its organic substance. The CXB possessed the same natural porous network system as that of the human. The CXB was cultured with the bone marrow stromal cells of rabit. When the marrow cells had integrated with the CXB, thus a new material was obtained. (CXB-BM), and was implanted sacro-spinal muscle of rabbit. The specimens were observed under phase microscope, light microscope and electronic scanning microscope. The results showed that: at the 2nd week after the implantation of CBX-BM composite material there began the new bone formation, and the rate of bone formation was increased with time. There was evident new bone formation after 24 weeks. The process of the new bone formation were quite similar to the composite graft of HAP red autogenous and marrow, but the former degraded faster and formed typical cancellous structure earlier. There was no new bone formation when CXB was implanted alone in the control. Both the mechanism of osteogenetic potential and its clinical application were discussed. PMID- 10374604 TI - [Clinical xenotransplantation: current progress, problems and solutions]. AB - Xenotansplantation has become a global focus because it may solve the formidable problems in allotransplantation, that is, the donor source. Hitherto clinical xenotransplantion has been in the stage of research with limited cases and unsatisfactory results. The difficulties which hinder the progress of xenotransplantation include: the ideal animal donor has not been found, it is rather difficult to control the rejections (hyperacute rejection, acute vascular rejection, perhaps acute cellular rejection and chronic rejection) after xenotransplantation compared with those after allotransplantation, some animal diseases might be transmitted to and do harm to human recipients, even the community. It is still unknown whether the functions of animal organs can substitute those of human organs permanently. Transgenic pigs on research and various measurements to suppress humoral and cellular immunity may be helpful in overcoming the problems of xenogeneic rejections. Animal diseases should be prevented, screened and treated, and animal models should be established to study the possibility of satisfactory working of animal organs in human body before clinical xenotransplantation is widely practised. PMID- 10374605 TI - [Study on the activity of fibroblast in hypertrophic scar]. AB - To determine the state of fibroblast during the process of development of hypertrophic scar (HS), 40 specimens of HS in different periods were collected. The expressions of prolifrating cell nuclear antigen (PCNA) and Ag-protein in nucleolar organizer regions (Ag NORs) as well as the content of total amino acids in the tissues were examined. The hypertrophic scar of 1st and 3rd month old, the expression of PCND and Ag NORs were the highest. In the 9th and 12th month old, althrough PCNA was nearly negative, but the expression of Ag NORs was low. The content of total amino acid was increased gradually as HS developed but the increase of amount of hydroxyproline was markedly slowed down in 9 month old HS. It was suggested that: (1) in the developing process of HS the proecess of overproliferation of fibroblasts was short and limitted in 1-3 months period in the process of wound lealing; (2) the synthesis of collagen was nearly stopped at 6 months, but that of other extracellular matrix such as fibronectin and proteoglycan might be continued to aggregate after 12 months. PMID- 10374606 TI - [Effect of crushing of sciatic nerve on neuron of lumbar spinal cord]. AB - In order to investigate the effect of nerve compression on neurons, the commonly used model of chronic nerve compression was produced in 48 SD rats. The rats were sacrificed in 1, 2, 3, 4, 5 and 6 months after compression, respectively. The number of neuron and ultrashruchure of alpha-motor neurons and ganglion cells of the corresponding spinal segment were examined. The results showed as following: After the sciatic nerve were crushed, the number of neuron and ultrastructure of alpha-motor neurons and ganglion cells might undergo ultrastructural changes, and even the death might occur. These changes might be aggravated as the time of crushing was prolonged and the compression force was increased. It was concluded that for nerve compression, decompression should be done as early as possible in order to avoid or minimize the ultructural changes of the neuron. PMID- 10374607 TI - [Repair of cartilage defect in joint with transplantation of cryopreserved homologous embryonic periosfeum of rabbits]. AB - In order to repair cartilage defect in joint with transplantation of cryopreserved homologous embryonic periosteum, 30 rabbits were used and divided into two groups. A 4 mm x 7 mm whole thickness cartilage defect was made in the patellar groove of femur of each rabbit. The homologous embryonic rabbit skull periosteum (ERSP), preserved in two-step freezing schedule, was transplanted onto the cartilage defect of joints of one group and autogenous periosteal graft was done in the joint defect of the other group. The knees were not immobilized, following operation and 16 weeks later, the newly formed tissue in the defects were assessed by gross observation, histochemical examination and biochemical analysis. The results showed that new hyaline-like cartilage was formed in the cryopreserved ERSP grafted knee, and had no significant difference from that of the knee receiving autogenous periosteal graft, but had significant difference from that of the fresh ERSP grafted knee and the non-grafted knee. Furthermore, the new hyaline-like cartilage had the biochemical characteristics of a fibrous cartilage. The conclusion was that this method might be feasible to repair articular cartilage defects. PMID- 10374608 TI - [Clinical uses of medical hyaluronic acid]. PMID- 10374609 TI - [Effects of different vascular beds on the microstructural components and intimal hyperplasia of autogenous vein grafts in dogs]. AB - In order to investigate the effect of vascular beds on the vascular wall of autogenously grafted vein, femoral veins were reversely placed in between the cut ends of collateral femoral arteries in 11 dogs with atraumatic technique. The grafted veins were covered with vivid muscle or skin respectively after being assured to be patent, and investigated by histomorphologic method and computerized image analysis technique at postoperative intervals of 1 week, 4 weeks and 16 weeks. The results showed that: 1. One graft developed pseudoaneurysm at 1 week, and two grafts were occulded in skin-covered group, whereas, no complications occurred in muscle-covered group. 2. Intimal thickening of grafts in skin-covered group was much more obvious than that in the muscle covered group (P < 0.05). 3. The relative contents of microstructural components of the graft wall showed no significant difference quantitatively between the two groups. So, the conclusion was: 1. Subcutaneous transplantation appeared to be a potential causative factor in inducing short-term excessive dilatation and long term intimal hyperplasia of vein graft. 2. Muscular covering is of priority in blood vessel graft. PMID- 10374610 TI - [A new method to repair artery injuries in extremities by phleboplasty of branched vein graft]. AB - In order to develope a new method to overcome the difficulties in anastomosis of blood vessels with different diameter, phleboplasty was utilized at the join point to expand the diameter of branched vein graft, with a funnel-shaped stoma formed consequently. After successfully experimented in fresh blood vessels in vitro, the method was practised clinically to repair injured arteries in extremities, with the outcome that phleboplasty of branched vein graft could enlarge the diameter by 1-1.25 times, and with satisfied effects in 3 clinic cases. So, the conclusion was that: phleboplasty of branched vein graft was a new effective and convinient method to repair injured arteries with different diameters. PMID- 10374611 TI - [Treatment of ischemia in lower extremities by primary arterization in situ of Vana saphena magna]. AB - In order to summarize the experience in the treatment of ischemic necrosis of lower extremities resulted from thrombotic occluded angittis, 15 cases were reported, which were treated by primary arterization in situ of V. Saphena magna. With a period of follow-up, 4-26 months on the average, it was found that symptoms in 14 cases were much allayed obviously, except 1 case with little relief. It suggested that primary arterization in situ of V. saphena magna could improve the circulation of the ischemic extremity rapidly without any influence of venous reflux. PMID- 10374612 TI - [Treatment of venous reflux disease of the leg by deep-venous valve reconstruction]. AB - In order to evaluate the effect and indication of three kinds of deep-venous valve reconstruction surgery, 62 cases with venous reflux disease of the leg had been treated from Jan. 1992 to Jun. 1996. All the patients had varicose vein and tingle in varying degrees, besides swelling in 30 cases, pigmentation in 28 cases, ulcer in 14 cases. The course of disease ranged from 1 to 30 years (averaged 14.6 years). METHOD: 14 cases were treated by ringing of the superficial femoral venous 1st valve, 1 case was treated by repairing of the superficial femoral venous 1st valve, 47 cases were treated by formation of substitute valve outside the popliteal vein. The symptoms of all the patients were alleviated with an average follow-up for 20 months. Fourteen cases with ulcer were healed compeletely and no recurrence. The conclusion were: 1. ringing and repairing of the venous valve were suitable for level I-II venous valvular incompetence. 2. formation of substitute valve outside the popliteal vein was suitable for level III-IV venous valvular incompetence or congenital valvular defect. 3. the width of the ringing material should be increased to 2 cm according to the pathological basis. 4. both femoral veins should be ringed in the treatment of primary valvular incompetence of double deep vein deformity. 5. the formation of substitute valve outside the popliteal vein was also available in the treatment of popliteal vein with many branches. PMID- 10374613 TI - [The comprehensive treatment of varicosis of lower extremity with chronic ulcer of leg]. AB - From Oct. 1993 to Dec. 1995, nineteen refractory cases with varicosis and chronic ulcer of lower limb were treated. The average age of these patients was sixty eight, the disease history was more than 20 years. The size of the ulcer of the leg ranged from the minimum of 10 cm x 8 cm to the maximum of 30 cm x 15 cm. All of them had once received saphenectomy and split skin graft without ulcer healing before they were admitted in our department. Both venography and ultrasonography showed superficial venous valve incompetence. The following comprehensive treatment was adopted. Firstly, myoplasty around popliteal vein was done. Secondly, phlebexairesis and phleborrhaphy were done for the variciform veins through minor incision. Then through debridement of the ulcer was performed. Delayed split skin graft was exerted one week later. The result showed that all the cases were successful: the ulcer was healed and there was no recurrence of varicosis. PMID- 10374615 TI - [Review on the treatment of superficial hemangioma]. PMID- 10374614 TI - [The clinical application of bridge by "Y" type vein]. AB - In order to solve the defect of blood vessel in tissue transplantation and complicated palmar amputation, bridge by "Y" type vein had been used from Jan. 1990 to Jul. 1996. Twenty-three cases were treated. In this series, there were 16 males and 7 females, with ages ranged from 10 to 42 years old. Six cases were the defect of lower legs anterior skin and tibia, 3 cases were the femur fracture with injury of femoral artery and tissue's defect, 2 cases were defect of five fingers, 12 cases were complicated palmar amputation. RESULT: 15 cases with tissue transplantation and 12 cases with limb replantation were all survival without infection or necrosis. After the following-up for 3 years (ranged from 1 to 5 years), the function of injured limbs were satisfactory, 19 patients had resumed their original work. So, to bridge by "Y" type vein is a good method for repairing the defect of blood vessels in tissue transplantation and complicated palmar amputation, but skilled microsurgery technique is required. PMID- 10374616 TI - [One stage repair and reconstruction for severe deep burns with compound tissue defects of upper limb]. AB - In order to solve the difficult problems of repair and reconstruction for severe deep burns with compound tissue defects of upper limb, 26 cases were treated with transplantation of compound tissue flap, vascularized by anastomosis of blood vessel or by vascular pedicle. Several kinds of reparative and reconstructive procedure could be performed simultaneously. Not only the tissue defect was repaired, but also the upper limb function was reconstructed in one stage operation. Owing to the presence of abundant vascular supply from the vascularized compound tissue and primarily closing the wounds, the anti-infection potency was high, then it was suitable for such conditions as fresh severe deep burn with infection and compound tissue defects. As a result, this technique provided the best chance to save upper limb from amputation. The duration required for treatment could be markedly shortened. All the cases successed. The long-term functional recovery was satisfactory. This method provided the possibility to solve effectively the difficult problem dealing with the severe deep burns with compound tissue defects of upper limb. PMID- 10374617 TI - [The treatment and rehabilitation of high-voltage electric burns]. AB - High-voltage electric burns is refractory with high rate of amputation (46%) in early stage and unfavorable functional recovery in later stage. Little breakthrough has so far been made in this respect. From Jan. 1985 to Jan. 1996, ninety-six cases with high-voltage burns were treated in our department. Seventy one cases of various tissue flap grafting were applied to treat early electric burns, among which sixty-four cases were successful. The amputation rate was reduced to 30%. Postoperatively, a long-term rehabilitation training at home was carried out. Most of them achieved a good appearance of the wounded sites and limbs and satisfactory ability to work or self-care. It was suggested that early thorough debridement of necrosis tissue, careful reservation of living tissue, appropriate choice of tissue flap and postoperative rehabilitation training were of great importance to achieve a good prognosis. PMID- 10374618 TI - [Causative ananlysis for redislocation after operative reduction of congenital dislocation of hip]. AB - Redislocation of the femoral head may be occured after its operative reduction in the congenital dislocation of the hip, therefore, it is greatly important to disclose the causes of the redislocation in order to avoid this every complication and improve the curative effect of this operation. Seven cases of redislocation from 106 cases (128 sides) of the congenital dislocation of the hip which had been reduced operatively were studied with relative measurements of the hip joints on roentgenogram, associated their pathologic conditions described in operation. The results showed that, in these cases, there were (0.843 +/- 0.692) cm upward displacement of the femoral head beyond the horizontal Y line, (68.86 +/- 0.692) degree of the femoral anteversion, the more lateral displacement of the femoral head compared to the opposite side and the acetabular index increasing up to (33.86 +/- 3.72) degree from (26.14 +/- 2.73) degree of the operative correction. These phenomena indicate that the redislocation after operative reduction of the femoral head in congenital dislocation of the hip is mainly related to four causes which include the existence of large pressure between acetabulum and femoral head, the uncorrected abnormal femoral anteversion, the residue of the pathologic tissues in the acetabulum and the reascending of the acetabular index having been corrected in operation. PMID- 10374619 TI - [Analysis of causes of revision after hip replacement]. AB - From 1974 to 1991, two hundred and sixty-four cases of hip replacement were performed. These cases composed of 150 cases of artificial femoral head prosthesis replacement and 114 cases of total hip replacement. Fifteen cases were revised after the first replacement in 7.4 years average (5-16 years). The revision rate was 5.7%. The causes of revision were loose or subside of prosthesis, wear and tear of acetabulum, dislocation of artificial hip joint, etc, which caused pain and dysfunction. The revision cases were followed up for 4.7 years average with good result. To prevent revision, The medully canal shonld not be too wide and in osteoporosis cases, bone cement was suggested to apply. The chondrium of acetabulum should be removed completely. PMID- 10374620 TI - [Prevention and treatment of postoperative complications following skin soft tissue expansion]. AB - Since 1987, One hundred and fifty-four patients suffered from alopecia, neck and facial scar, and nasal defect had been treated with skin soft tissue expansion. The incidence of complication was decreased markedly, compared to previons report which was 11.7%. Two cases of this group were given up this procedure. The lessous learned from these case were as following. Strictly evaluated the case according to the indication, examined the expander carefully, improved the techniques to inbed the expander and infilled the sailine, those of which could obtain satisfactory result. PMID- 10374621 TI - [The effect of blocking insulin-like growth factor-1 (IGF-1)-receptor system on tendon cell proliferation]. AB - The purpose of this study was to find some solutions to the problem of tendon cell proliferation control. Under the condition of in vitro culture, several materials including IGF-1 receptor antibody and mRNA antisense oligonucleotide were added to the culture medium to block the IGF-1-Receptor system. The effect of the material on the tendon cell proliferation was judged by cell count after incubation of 48 hours. The results showed that both IGF-1 Receptor antibody (IGF 1R alpha) and IGF-1 Receptor mRNA antisense oligonucleotide had negative effect on tendon cell proliferation (P < 0.01 and P < 0.05). These findings lead us to think that the above two materials could be used in the experiment of tendon adhesion preventing and living ready-made tendon producing. PMID- 10374622 TI - [Osteogenic characteristics of bone marrow and composite bone marrow transplantation. A review]. PMID- 10374623 TI - [Xenogeneic antigens and immune response in pig to man xenograft]. AB - Limitation of donor source for allograft makes the research on xenograft progress. Pig is regarded as one of the ideal donor animals. The major obstacle in xenograft is hyperacute rejection, which is caused by complements after they are activated by xenogeneic antigens combined with natural antibodies. It has been confirmed that alpha-Gal is the major target antigen, whose expression is incharged by alpha-1,3 galactosyltransferase (alpha-GT). The approaches to overcome hyperacute rejection against alpha-Gal included: immunoadsorption of xenogeneic natural antibodies, lysis of antigen by enzyme and genetic manupilation to obtain animal lack of alpha-GT. Besides alpha-Gal, there were other antigens binding to human serum antibody, such as gp65 and gp100, which was expressed on PAEC after induced by TNF, the A-like antigen. But their function was still unknown. It was debatable on the role of MHC in xenograft. Both direct and indirect pathway were involved in cellular response in xenograft. PMID- 10374624 TI - [The effects of clenbuterol on intramuscular collagen metabolism in denervated muscle]. AB - In order to explore the effects of clenbuterol on intramuscular collagen metabolism in denervated skeletal muscles, a randomized, double-masked and placebo-controlled group were studied. Seventy-one patients with complete function loss in muscularcutaneous nerve resulted from brachial plexus injury were administered clenbuterol or placebo 60 micrograms Bid for more than 3 months. Biopsies of the biceps brachia muscle were performed at the beginning and end of this study. The biopsied muscles were processed with anti-collagen I and IV immunohistochemical stains and image analysis as well. The result showed that the collagen proliferation of both type I and IV was much reducible in the clenbuterol-treated group than that of the placebo-treated group (P < 0.05). It was concluded that clenbuterol could inhibit partially the proliferation of intramuscular collagens in denervated skeletal muscle. PMID- 10374625 TI - [The experimental study of the effect of insulin and danshen on culture of fibroblast in vitro]. AB - To investigate the effect of Insulin and Danshen on anabolism and catabolism of collagen during the healing of wound, fibroblast which was cultured from human embryonic skin were divided into 3 groups: Insulin group, Danshen group and contral group. Each group was cultured for 2, 4 and 6 days, then the growth curve was established respectively. RESULTS: 1. The growth curve showed-insulin group > control group > Danshen group. 2. The rate of cell division in 3 groups was 19.6/1000, 2.5/1000, 3.77/1000 respectively. 3. The electron microscopic scanning showed that there was much of fibroid tissue surrounding the fibroblast cell in insulin group, but there was little fibroid tissue in Danshen group. The conclusion showed that insulin can accelerates proliferation of fibroblast and synthesis of collagen, but the effect of denshen was just on the contrary. PMID- 10374626 TI - [Therapeutic effects of ultrashort wave and He-Ne laser on experimental infection in skin flaps of rabbits]. AB - In order to investigate the therapeutic effects of ultrashort wave and He-Ne laser on experimentally infected skin flaps, 24 lower abdominal skin flaps on 24 rabbits were established, under each flap 1 ml of S. aureus (9 x 10(8) bacterials/ml) was injected respectively. Then, ultrashort wave and He-Ne laser were utilized respectively in 2 groups once a day for 6 days, with on treatment in two another two groups as control groups. After the period of treatment, the 4 groups were evaluated in the bacterial amounts, thickness of skin flaps and degree of infection. The results showed that the skin flaps in the treatment groups were much better than those in the control groups. It suggested that both the ultrashort wave and He-Ne laser are helpful in the treatment of infection of S. aureus in skin flaps. PMID- 10374627 TI - Comparison of binding affinities of omega-conotoxin and amlodipine to N-type Ca2+ channels in rat brain. AB - AIM: To compare the binding affinities of omega-conotoxin (CTX) and amlodipine to N-type Ca2+ channels in rat brains. METHODS: Whole rat brains were homogenized in HEPES buffer 50 mmol.L-1 (pH 7.4) and centrifuged at 40,000 x g to obtain the membrane-entriched fraction. 125I-omega-conotoxin (125I-omega-CTX) was used as a radioligand. Using radioligand binding assay Kd and Bmax values of the radioligand were determined by Scatchard analysis. The IC50 value for each drug was obtained from displacement experiments. RESULTS: No differences in Bmax values of 125I-omega-CTX binding sites between frozen and fresh tissues were observed. Values of Kd and Bmax of N-type Ca2+ channels were 0.02 +/- 0.01 nmol.L 1 and 1029 +/- 108 pmol/g protein, respectively. The pKi values of omega-CTX and amlodipine were 9.57 and less than 4, respectively. The pKi values of propranolol, prazosin, atropine, and histamine were very low. CONCLUSION: The binding affinity of the L-type Ca(2+)-antagonist amlodipine to N-type Ca2+ channels in the rat brain was very low. PMID- 10374628 TI - Enhancement of (-)-stepholidine on protein phosphorylation of a dopamine- and cAMP-regulated phosphoprotein in denervated striatum of oxidopamine-lesioned rats. AB - AIM: To study effects of (-)-stepholidine (SPD) on the phosphorylation of a dopamine- and cAMP-regulated phosphoprotein (DARPP-32) in the striatum of oxidopamine-lesioned rats. METHODS: The amount of dephospho-DARPP-32 was measured by a back-phosphorylation assay. RESULTS: In the striatum of control rats, SPD per se had no effect on the phosphorylation of DARPP-32, but it antagonized the decrease by 28% of dephospho-DARPP-32 induced by the D1 agonist SK&F-38393. In the denervated striatum of oxidopamine-lesioned rats, SPD decreased the amount of dephospho-DARPP-32 by 44%. The effect of SPD was completely counteracted by the concomitant administration of the D1 antagonist Sch-23390. CONCLUSION: SPD exhibits D1 agonistic action on DARPP-32 phosphorylation in the denervated striatum of oxidopamine-lesioned rats, but it acts as a D1 antagonist in normal striatum. PMID- 10374629 TI - Neuroprotective effects of poly (ADP-ribose) polymerase inhibitors in transient focal cerebral ischemia of rats. AB - AIM: To explore the role of poly (ADP-ribose) polymerase (PARP) in focal cerebral ischemia with reperfusion injury. METHODS: Male Wistar rats underwent 3.5-h of temporary middle cerebral artery occlusion by intraluminal suture. Infarction volume was showed with 2,3,5-triphenyltetrazolium chloride (TTC) staining and quantitated by image analysis system, neurologic scores were determined with a 0 5 grading scale. RESULTS: 3-Aminobenzamide (3-AB) 10 mg.kg-1 or nicotinamide (Nic) at 20 mg.kg-1 showed potent neuroprotective effects within 0-6 h, neurologic deficits were attenuated. With the increasing dose of PARP inhibitors, beneficial effects were compromised, particularly, administration of Nic 60 mg.kg 1 at the onset of reperfusion drastically accelerated brain damage. Phytomenadione, a selective inhibitor of mono (ADP-ribosyl) transferase, had little effect on infarction volume. CONCLUSION: Transient incomplete inhibition of PARP provides a neuroprotective effects against cerebral ischemia-reperfusion injury, with a relatively wide therapeutic window, whereas severe inhibition of this enzyme, especially in reperfusion phase, is detrimental. PMID- 10374630 TI - Enhancing effects of beta-endorphin on glutamate neurotoxicity. AB - AIM: To study the effect of beta-endorphin (beta-End) on monosodium glutamate (MSG)-induced neurotoxicity (GNT). METHODS: Image analysis of neuronal areas and determination of mitochondrial membrane protein-bound Ca2+ and intracellular free Ca2+ ([Ca2+]i) were used. RESULTS: beta-End aggravated MSG-induced neuronal injury in arcuate nucleus of hypothalamus in a dose-dependent manner in the range from 0.5 to 5.0 mg.kg-1. MSG-induced increase in mitochondrial membrane protein bound Ca2+ was enhanced when treated with beta-End 2 g.L-1. MSG-induced elevation in [Ca2+]i in single neuron was also augmented from 320 +/- 84 to 589 +/- 78 nmol.L-1 by the treatment with beta-End 2 g.L-1. CONCLUSION: beta-End enhanced GNT via aggravating the disruption of intracellular Ca2+ homeostasis induced by MSG. PMID- 10374631 TI - Belladonna alkaloids-induced behavioral changes and amnesia on open-field and step-through in 18-, 28-, and 38-day-old mice. AB - AIM: To study the age-related changes of atropine (Atr), scopolamine (Sco), anisodine (AT3), and anisodamine (Ani) on behaviors and memories. METHODS: The behaviors and memories were measured with open-field test and step-through task. M-cholinergic receptors were determined by [3H] quinuclidinyl benzilate ([3H] QNB). RESULTS: During acquisition session (d 1) the 18-, 28-, and 38-d-old mice pretreated with Atr, Sco, and AT3 (0.02, 0.2, 2, or 20 mg.kg-1, i.p.) in open field test showed increase in walking counts by 26%-42%, but decrease in rearing, grooming, and defecating counts for 50%-92%, 67%-100%, and 75%-100%, respectively. On recall session (d 2) the walking and rearing behaviors in the 18 and 28-d-old mice receiving Atr, Sco, and AT3 on d 1 were higher than those in the mice receiving saline. But a lower grooming behavior on d 2 was found in the mice receiving the drugs on d 1. On d 1 Ani 20 mg.kg-1 reduced the rearing behavior by 50% in 18-d-old mice and defecation by 33%-36% in 18- and 28-d-old mice. All the 4 belladonna alkaloids increased the number of avoidance-response errors and decreased the retention latencies in step-through task. Bmax of [3H] QNB binding sites in frontal cortex and hippocampus regions in the 38-d-old mice increased 7% and 23% vs in the mice of 18 d of age, respectively. CONCLUSION: 1) The effects of the belladonna alkaloids on behaviors and memories in adult mice were weaker than those in young mice. 2) The belladonna alkaloids-induced amnesia on passive avoidance-response in step-through was more sensitive than behavioral changes and amnesia on open-field. 3) According to the lowest effective doses which insulted the behaviors or memories in young mice, Sco was about 10, 100, and 1000 times more potent than Atr, AT3, and Ani, respectively. PMID- 10374632 TI - Effects of dl-3-n-butylphthalide on regional cerebral blood flow in right middle cerebral artery occlusion rats. AB - AIM: To study the effect of dl-3-n-butylphthalide (NBP) on regional cerebral blood flow (rCBF) in forcal cerebral ischemia rats. METHODS: In chloral hydrate anesthetized rat, the proximal portion of right middle cerebral artery (RMCA) was occluded, and H2 needle electrode was implanted in right striatum. rCBF was monitored in striatum using hydrogen clearance method. RESULTS: Ten min after RMCA occlusion (RMCAO), NBP (5, 10, 20 mg.kg-1 i.p.) markedly increased rCBF to striatum (P < 0.01). When NBP was given i.p. 40 min after RMCAO, the increasing effect on rCBF was also observed (P < 0.05). However, when NBP was injected i.p. 60 min after RMCAO, the increasing effect of NBP on rCBF was not found. In NBP pretreated (i.p. 40 min before RMCAO) group, rCBF in striatum measured at different time points of 30, 60, 90, 120, 150, and 180 min after RMCAO were increased by 97%, 107%, 136%, 211%, 173%, and 317%, respectively, compared with the percentages of vehicle group. The potency of the effect of Nim (0.5 mg.kg-1 i.p.) was similar to that of NBP (10 mg.kg-1 i.p.). CONCLUSION: NBP pre-treatment or post-treatment markedly enhanced the rCBF to striatum in RMCAO rats. PMID- 10374633 TI - Construction of an inducible nitric-oxide synthase gene transferring vector mediated by retrovirus. AB - AIM: To construct an inducible nitric-oxide synthase (iNOS) gene transferring vector mediated by retrovirus. METHODS: Recombinant DNA and polymerase chain reaction (PCR) amplification techniques were used. RESULTS: With 2 steps of molecular cloning, the full-length cDNA encoding macrophage iNOS was isolated from plasmid pKSiNOS and subcloned into intermediate vector pSP72, adjusting the restriction enzyme sites in both 5'- and 3'-flanking ends of insert fragment. The retroviral vector pLNCXiNOS which contains iNOS coding region, cytomegalovirus promoter and neomycin resistance (neor) gene was further constructed. The authenticity of insertion size and orientation of iNOS sequence was verified by restriction mapping and PCR analysis with iNOS gene-specific primers. CONCLUSION: Retroviral expression vector carrying iNOS fragment is obtained, which provides a material to establish a model of iNOS gene-modified neurons. PMID- 10374634 TI - Effects of huperzine A on nucleus basalis magnocellularis lesion-induced spatial working memory deficit. AB - AIM: To study the effects of huperzine A on nucleus basalis magnocellularis (NBM) lesion-induced spatial working memory impairment. METHODS: A delayed-non-match-to sample radial arm maze task was used to study spatial working memory. The choline acetyltransferase (ChAT) activity was determined by the conversion of [3H]acetyl CoA to [3H]ACh. RESULTS: Unilateral NBM lesion by kainic acid 0.02 mumol impaired rat's ability to perform this working memory task as evidenced by fewer correct choices after different delay intervals and more total errors to complete the task. This behavioral impairment associated with a decrease in the activity of ChAT by about 40% in the ipsilateral cerebral cortex. Huperzine A (0.2 mg.kg-1 i.p. 30 min before testing) ameliorated this spatial working memory impairment. Physostigmine (0.2-0.3 mg.kg-1 i.p. 20 min before testing) also attenuated the NBM lesion-induced memory deficit. CONCLUSION: The integrity of NBM is critical for spatial working memory processing, and this working memory impairment induced by NBM lesion can be ameliorated by huperzine A and physostigmine. PMID- 10374635 TI - Inhibitory effect of antisense basic fibroblast growth factor oligonucleotides on proliferation of cultured aortic smooth muscle cells induced by angiotensin II in SHR rats. AB - AIM: To study the effect of antisense basic fibroblast growth factor (bFGF) oligonucleotides (ODN) transfection on the growth of cultured aortic smooth muscle cells (SMC) in spontaneously hypertensive rats (SHR). METHODS: Using cationic liposome-mediated method, antisense bFGF ODN were introduced into SMC, bFGF gene expression was detected by Northern blotting, cell hyperplasia was evaluated by [3H] thymidine incorporation and cell counting. RESULTS: Transfection of antisense bFGF ODN (5 mumol.L-1) almost completely inhibited enhanced bFGF mRNA expression and inhibited cell proliferation induced by angiotensin II (Ang 1 mumol.L-1). In basal state and Ang-stimulated state, [3H]thymidine incorporation was inhibited by 26.5% (P < 0.01) and 42.0% (P < 0.01) and cell number was inhibited by 17.3% (P < 0.01) and by 22.2% (P < 0.01), respectively. CONCLUSION: The transfection of antisense bFGF ODN into cultured SMC effectively suppressed bFGF mRNA expression and inhibited the SMC proliferation induced by Ang. PMID- 10374636 TI - Proliferation of aortic smooth muscle cells and renin-angiotensin system in SHR rats. AB - AIM: To study the relationship between the enhanced proliferation and renin angiotensin system (RAS) of aortic smooth muscle cells (ASMC) from SHR rats. METHODS: To measure the effects of angiotensin II (Ang), captopril (Cap), saralasin (Sar) on proliferation, Ang and angiotensin converting enzyme (ACE) levels in cultured ASMC from WKY and SHR rats. RESULTS: Ang was a bifunctional growth factor, which induced SHR ASMC hyperplasia in 2% FCS-RPMI 1640 medium, but not in serum free (SF)-medium. SHR ASMC had stronger proliferative ability compared with WKY while SHR ASMC RAS was activated. Enhanced proliferation of SHR ASMC and ACE activity were obviously inhibited by long-term treatment (4-wk) of both Cap and Sar, while Ang content decreased in Cap treatment group and increased in Sar treatment group. The antiproliferative effect of Cap and Sar on SHR ASMC was stronger than that on WKY. SHR, WKY ASMC RAS were not influenced by short-term (24 h) treatment of Cap. CONCLUSION: Long-term treatment of Cap and Sar suppressed SHR ASMC growth through inhibition of Ang generation or blockade of Ang binding to its receptor. PMID- 10374637 TI - Effects of norepinephrine and isopentenyladenosine on Na+/Ca2+ exchange currents in isolated guinea pig ventricular myocytes. AB - AIM: To study the effects of norepinephrine (NE) and isopentenyladenosine (Iso) on Na+/Ca2+ exchange currents and the receptor mechanism. METHODS: The quasi steady state current-voltage relationship from the isolated guinea pig ventricular myocytes was measured using whole-cell voltage-clamp techniques with a ramp pulse protocol. RESULTS: At potential of +50 mV, NE 0.005, 0.05, and 5 mumol.L-1 increased the Ni(2+)-sensitive current by 29% +/- 9%, 72% +/- 11%, and 124% +/- 31.4%, respectively; Iso 1.5, 150, and 1500 nmol.L-1 caused increases in the Ni(2+)-sensitive current by 2.8% +/- 2.8%, 56% +/- 13%, and 102% +/- 12%, respectively. Propranolol 10 mumol.L-1 completely inhibited the current changes induced by NE and Iso while phentolamine 50 mumol.L-1 showed no effects. CONCLUSION: NE and Iso increased the Na+/Ca2+ exchange currents via stimulation of cardiac beta-adrenoceptor. PMID- 10374638 TI - Effects of metallothionein on action potentials of anoxic and reoxygenated papillary muscles of guinea pigs. AB - AIM: To study anti-arrhythmic effects of metallothionein (MT). METHODS: Standard microelectrode technique was used to study the effects of MT on action potentials (AP) in anoxic and reoxygenated papillary muscles of guinea pigs. RESULTS: MT (0.02 mmol.L-1) had no effects on AP of the normal papillary muscles; when the muscles were exposed to ischemic solution without MT, there was a marked shortening of action potential duration (APD) at 20%, 50%, and 90% of repolarization (APD20, APD50, APD90) from 82 +/- 7 to 37 +/- 7, 131 +/- 35 to 63 +/- 11, and 167 +/- 12 to 100 +/- 19 ms, respectively, (P < 0.01); and an obvious reduction of resting potential (RP), action potential amplitude (APA), and the maximal upstroke velocity of phase 0 (Vmax) from -92 +/- 9 to -63 +/- 12 mV, 135 +/- 13 to 80 +/- 8 mV, and 286 +/- 55 to 164 +/- 42 V.s-1, respectively (P < 0.01). However, in the presence of MT, the AP parameters (RP, APA, and Vmax) changed from -63 +/- 2 to -82 +/- 1 mV, 80 +/- 8 to 104 +/- 25 mV, and 164 +/- 42 to 237 +/- 43 V.s-1, respectively, (P < 0.01), except that APD20, APD50, and APD90 shortened further from 37 +/- 7 to 12 +/- 3, 63 +/- 11 to 28 +/- 7, and 100 +/- 19 to 82 +/- 11 mV, respectively (P < 0.01). MT decreased the incidence of automaticity during reoxygenation from 91% to 33%. CONCLUSION: MT possesses a calcium regulatory property. PMID- 10374639 TI - Effects of verapamil on down-regulation of norepinephrine-induced beta adrenoceptors in cultured rat cardiomyocytes. AB - AIM: To determine whether verapamil (Ver) inhibits norepinephrine (NE)-induced beta adrenoceptors down-regulation in cultured rat cardiomyocytes. METHODS: [3H] Dihydroalprenolol (DHA) radiobinding assay was used to measure beta adrenoceptor density, fluorescent indicator Fura 2-AM was used to estimate levels of free cytosolic calcium ([Ca2+]i). RESULTS: Ver reduced [Ca2+]i and increased beta adrenoceptor density, NE increased [Ca2+]i and reduced beta adrenoceptor density of cultured cardiomyocytes, these effects were time and concentration dependent. Ver inhibited the above effects of NE. CONCLUSIONS: Ver increased beta adrenoceptor density and inhibited NE-induced beta adrenoceptor down-regulation of cardiomyocytes. PMID- 10374640 TI - Mobilization of intracellular Ca2+ modulates activation of Na+/H+ exchange in thrombin-stimulated platelets. AB - AIM: To study the relationship between intracellular calcium translocation and activation of Na+/H+ exchange in thrombin-stimulated platelets. METHODS: Intracellular Ca2+ ([Ca2+]i) and pH (pHi) were measured by a dual wavelength fluorophotometer with Fura-2 and pH-sensitive probe BCECF. RESULTS: Thrombin 0.1 IU.L-1 elicited an increase in platelet [Ca2+]i and pHi, the maximal increase in [Ca2+]i occurred earlier than the rise in pHi. In Na(+)-free buffers, the Na+/H+ exchange was markedly suppressed without affecting the elevation of [Ca2+]i; while intracellular acidification with nigericin 1 mg.L-1 inhibited the increment of [Ca2+]i. Blockade of Ca(2+)-influx with egtazic acid (EGTA) did not affect cytosolic alkalinization. Depletion of intracellular Ca2+ store with ionomycin in the presence of EGTA, no increment in pHi was observed, the basal value of pHi was even more acidic, this response of pHi to thrombin was rehabilitated after refilling of intracellular Ca2+ store with extracellular Ca2+ 1 mmol.L-1. CONCLUSION: Intracellular Ca2+ mobilization modulated activation of Na+/H+ exchange, which required an effective increment of [Ca2+]i. PMID- 10374641 TI - Blocking effects of phentolamine on L-type calcium current and ATP-sensitive potassium current in guinea pig ventricular myocytes. AB - AIM: To study the effect of phentolamine on L-type calcium currents (ICa) and ATP sensitive K+ currents (IK,ATP) in ventricular myocytes. METHODS: ICa and IK,ATP were observed using patch clamp techniques in whole-cell recording configuration. RESULTS: Phentolamine reduced ICa of ventricular myocytes in concentration dependent and voltage-independent manners. Phentolamine 5, 25, and 100 mumol.L-1 decreased ICa from 370 +/- 99 nA to 310 +/- 95 nA (17% block, n = 6, P < 0.01), from 230 +/- 98 nA to 180 +/- 73 nA (23% block, n = 5, P < 0.05), and from 293 +/ 66 nA to 206 +/- 44 nA (30% block, n = 5, P < 0.01), respectively, without affecting the current-voltage relationship. Prazosin 100 mumol.L-1 and yohimbine 100 mumol.L-1, which were specific blockers of alpha 1 and alpha 2 adrenoceptors respectively, did not show the inhibitory effect on ICa. Phentolamine 100 mumol.L 1 also inhibited the IK,ATP induced by 2, 4-dinitrophenol (DNP) at 0 mV from 3.2 +/- 0.6 nA to 0.8 +/- 0.5 nA (75% block, n = 4, P < 0.01). CONCLUSION: Phentolamine directly inhibits ICa and IK,ATP in guinea pig ventricular myocytes. PMID- 10374642 TI - Inhibitory effects of nimodipine on platelet aggregation and thrombosis. AB - AIM: To study the inhibitory effects of nimodipine (Nim) on rat platelet aggregation and arterial thrombosis in vivo. METHODS: The aggregation rate of platelets induced by ADP and inhibition rate of Nim were measured by the change of light transmission. Effect of Nim on arterial occlusion time was measured by electric stimulation. Effect of Nim on the contents of 6-keto-PGF1 alpha and TXB2 in serum was measured by radioimmunoassay. RESULTS: Nim 4.5, 9, 18, and 36 mg.kg 1.d-1 ig for 4 d restrained the platelet aggregation. The IC50 (95% confidence limits) was 26 (9-44) mg.kg-1. Nim 4.5, 9, and 18 mg.kg-1.d-1 ig for 4 d markedly prolonged the time of thrombotic occlusion in carotid artery induced by electric stimulation. Nim 9 and 18 mg.kg-1.d-1 improved the imbalance of 6-keto-PGF1 alpha/TXB2 in serum after thrombosis. CONCLUSION: Nim was a potent inhibitor of platelet aggregation, which was partially concerned with the improved balance of 6-keto-PGF1 alpha/TXB2. PMID- 10374643 TI - Skeletal effects of constant and terminated use of sodium risedronate in ovariectomized rats. AB - AIM: To study the skeletal effects of constant and terminated use of sodium risedronate (Ris) treatment in the ovariectomized (Ova) rats. METHODS: Ris 5 micrograms.kg-1, s.c., twice a wk. The proximal tibial metaphysis (PTM) were processed undecalcified for quantitive bone histomorphometry. RESULTS: (1) Placebo-treated (normal saline) Ova rats were characterized by decreased trabecular area (TA) on d 60, d 81, and d 150 compared with aging controls, and bone resorption was over formation with high bone turnover. (2) Ova rats were treated with Ris for 60, 81, and 150 d (Ris-on) increased. (TA 217%, 108%, and 101%) respectively, vs Ova rats and depressed bone turnover indices to aging control level, but bone mass did not maintain at high level in 150-d group as in the early stage. (3) Ova rats were pretreated with Ris for 60 d and then terminated (Ris-on/off), followed by sequential sacrifice of rats on 21 and 90 d. Withdrawal on 21 d showed the same results as the match-age Ris-on group. Withdrawal on 90 d still maintained cancellous bone mass at a high level vs 150 d Ris-on groups (+26%) and aging control group (+27%). CONCLUSION: Regimen of Ris 60 d on then 90 d off prevented the development of osteoporosis in Ova rats. PMID- 10374644 TI - Inhibitory effects of procainamide on rabbit platelet aggregation and thromboxane B2 production in vitro. AB - AIM: To study the influences of procainamide (PA) on thrombin-induced rabbit platelet aggregation and thromboxane B2 (TXB2) production in vitro. METHODS: Turbidimetry and radioimmunoassay were used. RESULTS: PA 8.5, 34, 136, and 544 mumol.L-1 inhibited thrombin-induced platelet aggregation and TXB2 production, and the inhibitory rates were 45% +/- 37%, 48% +/- 32%, 88% +/- 23%, 92% +/- 15% and 53% +/- 24%, 65% +/- 26%, 90% +/- 6%, 95% +/- 6%, respectively. There was positive correlation between PA concentration and efficiency of inhibition of platelet aggregation and TXB2 production, and also between the inhibition % of platelet aggregation and that of production of TXB2. The three linear equations and main parameters were Y = 0.2075X-4.9157, r = 0.9985; Y = 0.9546X-34.6724, r = 0.9921; Y = 0.8202X + 19.7062, r = 0.9921. CONCLUSION: PA inhibited thrombin induced platelet aggregation and TXB2 production in rabbits. PMID- 10374645 TI - Glutathione-related enzyme activities in human fetal adrenal, liver, and kidney. AB - AIM: To understand the capacity of fetal adrenal to catalyze reaction metabolites. METHODS: Subcellular fractions were prepared by differential centrifugation in fetal adrenal and liver. Glutathione (GSH)-transferase, reductase, and peroxidase were measured. RESULTS: The mean values (mumol.min-1/g protein) of GSH-transferase activities in adrenal microsome (112 +/- 34), mitochondria (62 +/- 35), and cytosol (191 +/- 89) were 373%, 270%, and 167%, respectively, higher than those in the corresponding fractions of fetal liver. Adrenal microsomal GSH-transferase was positively correlated with adrenal microsomal P-450 (r = 0.821, P < 0.01), and with adrenal microsomal aminopyrine N demethylase (r = 0.829, P < 0.01). The GSH reductase contents (mumol.min-1/g protein) in adrenal mitochondria (24 +/- 14), and in S9 (36 +/- 15) were almost 5 times higher, compared with that in liver. Selenium-dependent GSH peroxidase was present in all the adrenal. CONCLUSION: Fetal adrenal, with greater capacities than those of liver in detoxifying reaction, may act as a drug-metabolizing organ during development. PMID- 10374646 TI - Dual effects of 5-hydroxytryptamine on stable analogue of thromboxane A2-induced aggregation and release reaction in rabbit platelets. AB - AIM: To study the effects of 5-hydroxytryptamine (5-HT) on stable analogue of thromboxane A2 (STA2)-induced platelet shape, aggregation, and release reaction. METHODS: Platelet shape change and aggregation were quantified by the light transmission through platelet-rich plasma (PRP). Release reaction was evaluated by the amount of ATP in the medium and cytosolic-free Ca2+ was measured by fluorescent imaging. RESULTS: (1) STA2 0.3-3 mumol.L-1-induced shape change followed by aggregation. When STA2 1 or 3 mumol.L-1 was added to PRP, the release reaction was occurred. Pretreatment of PRP with 5-HT 3 mumol.L-1, the shape change by STA2 was abolished and the aggregation by STA2 0.3 mumol.L-1 was enhanced (P < 0.01), STA2 1 or 3 mumol.L-1-induced aggregation was not affected, but the release reaction was partially suppressed (P < 0.01). (2) STA2 0.3 mumol.L-1-induced [Ca2+]i elevation was further increased by 5-HT pretreatment, but the [Ca2+]i mobilizations by STA2 3 mumol.L-1 was decreased by 5-HT, especially the peak level. (3) The aggregation without release reaction was increased from 3.4 +/- 2.1 to 25.6 +/- 1.8% (P < 0.01) with 10 s interval and the enhancement was declined with the prolongation of the intervals. The aggregation with release reaction was not affected by changing the intervals, but the release reaction was decreased in the same treatment. CONCLUSION: The dual effects of 5 HT on STA2-induced aggregation and release reaction and the molecular mechanism of this effect was probably through the regulative action of 5-HT on [Ca2+]i mobilization by STA2. PMID- 10374647 TI - Safety evaluation of peptides and proteins. PMID- 10374648 TI - Cognitive functions in schizotypal personality disorder. AB - OBJECTIVE: Schizophrenia spectrum subjects have cognitive deficits in a variety of domains. Schizotypal personality disordered (SPD) subjects do not have many of the confounds seen in schizophrenic patients, but may have the same pattern of cognitive deficits in attention and executive functioning. HYPOTHESIS: We hypothesized that SPD subjects would have impairments on measures of attention, abstract reasoning, cognitive inhibition, working memory and verbal recognition memory when compared to normal subjects, and that these deficits would be intermediate to those observed in schizophrenic patients. METHOD: SPD subjects (N=20) were compared to age-, gender- and education-matched schizophrenic patients (N=20) and normal comparison subjects (N=20) on a battery of cognitive measures. RESULTS: The data were analyzed using standard statistical methods, including effect sizes. Using a conservative alpha level of 0.01, schizophrenic patients had deficits on many of these measures compared to normal subjects. Although the SPD subjects did not significantly differ from normal comparison subjects at the p < 0.01 level, there were trends (p < 0.019-0.028) toward impairment on measures of working memory and general intellectual functioning. On further effect size analyses, SPD subjects performed intermediate to normals and schizophrenic patients on measures of attention, abstract reasoning, cognitive inhibition, verbal working memory, recognition memory, and general intellectual functioning, with moderate to large effect sizes separating groups. CONCLUSIONS: These results suggest that SPD subjects have possible widespread cognitive deficits that are of lesser magnitude than those observed in schizophrenic patients. PMID- 10374649 TI - Impairment in visual-spatial function in catatonia: a neuropsychological investigation. AB - Catatonia is a psychomotor syndrome with motor and behavioral abnormalities which may be due to alterations in fronto-parietal cortical function. We therefore investigated neuropsychological tasks (attention, executive, visual-spatial, working memory) associated with frontal and parietal cortical function. Thirteen catatonic patients, diagnosed as catatonic according to criteria by Rosebush and Bush, were compared with 13 psychiatric non-catatonic controls (matched with regard to underlying psychiatric diagnosis, age, sex, and medication), and 13 age and sex-matched healthy controls. Catatonics showed significantly poorer performances and different neuropsychological intercorrelation patterns in visual spatial object perception (VOSPobject) than psychiatric and healthy controls. In addition, we found significant correlations between catatonic symptoms, visual spatial abilities, and attentional measures (i.e., d2, CWI). Catatonia was characterized by specific visual-spatial deficits which are related to attentional abilities and right parietal cortical function. The data suggest attentional-motor and fronto-parietal dysfunction in catatonia, a conclusion which should be considered as preliminary, however, due to the small sample size. PMID- 10374650 TI - Failures of automatic and strategic processing in schizophrenia: comparisons of event-related brain potential and startle blink modification. AB - Noises elicit startle blinks that are inhibited when immediately (approximately 100 ms) preceded by non-startling prepulses, perhaps reflecting automatic sensory gating. Startle blinks are facilitated when preceded by prepulses at longer lead intervals, perhaps reflecting strategic processes. Event-related brain potentials (ERPs) and startle blinks were used to investigate the well-documented prepulse inhibition failure in schizophrenia. Blinks and ERPs were recorded from 15 schizophrenic men and 20 age-matched controls to noises alone and to noises preceded by prepulses at 120 (PP120), 500 (PP500) and 4000 ms (PP4000) lead intervals. Neither blinks nor any of the ERP components elicited by the noise alone differentiated schizophrenics from controls, although responses to noises were modified by prepulses differently in the two groups. With the N1 component of the ERP, patients showed normal inhibition but lacked facilitation, and with P2, patients lacked inhibition, but showed normal facilitation. With reflex blinks and P300, inhibition was seen in both groups, but no facilitation. These results suggest that different neural circuits are involved in blink and cortical reflections of startle modification in schizophrenics and controls, with both automatic and strategic processes being impaired in schizophrenia. PMID- 10374651 TI - Four behavioural syndromes of schizophrenia: a replication in a second inner London epidemiological sample. AB - In a previous large epidemiological survey of patients with strictly defined schizophrenia in the London borough of Camden, we extracted four behavioural syndromes (Social withdrawal, Thought disturbance, Anti-social behaviour and Depressed behaviour) by factor analysis of MRC Social Behaviour Schedule (SBS) data. These syndromes had significant differential relationships to symptoms assessed using the Manchester Scale (MS), symptom-derived syndromes, and social functioning variables. A second inner-London epidemiological survey of schizophrenia in South Westminster using identical methodology found the same four behavioural syndromes with identical core component items. The same four behavioural syndromes were extracted, whether applying strict Feighner diagnostic criteria (n=112) or broader DSM-III-R criteria (n=198). The four syndromes extracted from the Feighner positive sample showed relationships to symptoms and social functioning variables similar to those found in the original Camden study. However, the symptom-derived factors were not the same and did not conform to the three recognised symptom-based syndromes of schizophrenia. This successful replication suggests that assessment of the four behavioural syndromes of schizophrenia offers a different perspective on disability and a potentially relevant measure in clinical practice, clinical trials and studies of the neuropsychology and pathophysiology of schizophrenia. PMID- 10374652 TI - Meta-analysis of brain size in bipolar disorder. AB - A recent meta-analysis concluded that patients with schizophrenia have reduced cerebral volume, and this finding has been used to implicate neurodevelopmental events in the etiology of this disorder. Since bipolar-disorder patients and schizophrenia patients have some similar brain abnormalities, it was of interest to meta-analytically review the literature on brain size in bipolar disorder. Only seven studies met the inclusion/exclusion criteria for our meta-analysis, but none reported the brain size differences between the bipolar patients and the controls to be statistically significant. The composite effect size was a negligible 0.04 (95% CI: -0.17 to 0.25) and statistically not significantly different from 0.0 (no effect). Thus, it appears that bipolar disorder is not associated with the same cerebral volume reductions noted in schizophrenia. Implications for hypotheses regarding the etiology of the two disorders are discussed. PMID- 10374653 TI - Investigating theory of mind in schizophrenia: influence of verbalization in disorganized and non-disorganized patients. AB - The ability to attribute intentions to others was studied in 13 disorganized and 13 non-disorganized schizophrenic patients, 13 depressed and 13 normal controls. Subjects were asked to complete 28 comic strips requiring theory of mind skills by choosing one out of three answer cards. The answer cards were simple pictures in a first condition and short sentences in a second condition. This study, which used the cognitive neuropsychological approach, underlies the existence of a link between disorganization patterns in schizophrenia and a deficit in the attribution of intentions to others, independently of the pictorial or verbal form of the mode of answering. In addition, results show that the non disorganized schizophrenic group, depressed and normal controls perform similarly in both pictorial/verbal conditions. The influence of the absence/presence of verbal material on a task investigating theory of mind in schizophrenia is discussed. PMID- 10374654 TI - Application of pharmacogenetics to psychotic disorders: the first consensus conference. The Consensus Group for Outcome Measures in Psychoses for Pharmacological Studies. PMID- 10374655 TI - Gender differences in incidence and age at onset of DSM-III-R schizophrenia. Preliminary results of the Northern Finland 1966 birth cohort study. PMID- 10374656 TI - The association of neuropsychological deficits to clinical symptoms in first admission subjects with psychotic disorders. PMID- 10374657 TI - Low nerve growth factor plasma levels in schizophrenic patients: a preliminary study. PMID- 10374658 TI - Fast magnetic resonance imaging techniques. AB - This article reviews fast magnetic resonance (MR) techniques currently used for body imaging. Improvements in gradient performance have made very short repetition and echo times on clinical scanners feasible, thus enabling subsecond image acquisition. The article provides a fundamental overview of the technical aspects from the concept of k-space and k-space segmentation technique, fast MR imaging techniques including fast spin echo, fast gradient echo with or without magnetization preparation to echo planar and hybrid techniques. The article also addresses the use of different fat suppression techniques in MR imaging of the body and improvements in coil technology to obtain faster images and higher signal-to-noise. PMID- 10374659 TI - Fast magnetic resonance imaging of the heart. AB - Fast MR imaging techniques have multiple applications for evaluation of cardiac disease. Cine MRI and MR tagging have been shown to be highly accurate and reproducible in evaluating regional and global myocardial function. Segmented k space cine MRI and echo-planar imaging (EPI) can considerably improve time efficiency and thereby the clinical utility of these techniques. Double IR fast spin-echo sequences enable breath-hold acquisition of T2 weighted MRI with good suppression of the blood signal. Myocardial perfusion can be assessed with fast dynamic MRI after administration of contrast media. Multi-shot EPI improves temporal resolution and also provides full coverage of the left ventricle. Substantial progress has been made in respiratory gated 3D coronary artery MR angiography with navigator echoes. The newer approaches for coronary arterial imaging including breath-hold three-dimensional segmented EPI and high resolution spiral MRI may further improve clinical usefulness of coronary MR angiography. Assessment of coronary blood flow and flow reserve with phase contrast MRI has the potential for the non-invasive evaluating of the presence and significance of stenosis in the native coronary artery and bypass grafts. Fast cardiac MRI may emerge as a cost effective modality for comprehensive assessments of both cardiac morphology, function, blood flow and perfusion. PMID- 10374660 TI - Fast magnetic resonance imaging of the lung. AB - The impact of fast MR techniques developed for MR imaging of the lung will soon be recognized as equivalent to the high-resolution technique in chest CT imaging. In this article, the difficulties in MR imaging posed by lung morphology and its physiological motion are briefly introduced. Then, fast MR imaging techniques to overcome the problems of lung imaging and recent applications of the fast MR techniques including pulmonary perfusion and ventilation imaging are discussed. Fast MR imaging opens a new exciting window to multi-functional MR imaging of the lung. We believe that fast MR functional imaging will play an important role in the assessment of pulmonary function and disease process. PMID- 10374661 TI - Functional imaging of the kidneys with fast MRI techniques. AB - Availability of faster and stronger gradient systems have given rise to a multitude of fast MRI data acquisition strategies which have tremendously increased the scope of MRI applications. These have led to the realization of long desired comprehensive approaches to evaluate anatomy and function using a single modality. In this work, we describe some of our own experiences with functional evaluation of the kidneys using MRI. Examples that suggest the feasibility of comprehensive approaches for evaluation of renal disease are also provided. We also introduce BOLD renal MRI, a method that may allow basic understanding of human renal physiology and pathophysiology in a way that has not been previously possible. PMID- 10374662 TI - Quiz case 4. Churg-Strauss syndrome. PMID- 10374663 TI - MR imaging of pulmonary parenchyma with a half-Fourier single-shot turbo spin echo (HASTE) sequence. AB - OBJECTIVE: To evaluate the utility of a half-Fourier single-shot turbo spin-echo sequence (HASTE) at depicting lung parenchyma and lung pathology. METHODS AND PATIENTS: A HASTE sequence was applied to five normal volunteers and 20 patients with various pulmonary disorders to depict the lung parenchyma. Images were acquired with ECG-triggering and breath-holding. In three volunteers, signal intensity measurements from lung parenchyma were performed using four sequences: (a) HASTE; (b) conventional spin echo; (c) fast spin echo; and (d) gradient echo. T2 maps were produced using the HASTE acquisition. RESULTS: Minimal respiratory or cardiac motion artifacts were observed. The signal-to-noise ratios from lung parenchyma were 27.8 +/- 5.4, 22.0 +/- 3.0, 15.3 +/- 0.9, and 6.0 +/- 1.9 for HASTE, spin-echo, fast spin-echo, and gradient echo sequences, respectively. The scan time for HASTE was 302 ms for each slice. The T2 values in the right lung and the left lung were 61.2 +/- 4.1 and 79.1 +/- 8.9 ms in systole and 92.6 +/- 5.8 and 97.5 +/- 12.2 ms in diastole, respectively (P < 0.05 diastole versus systole). The HASTE sequence demonstrated clearly various pulmonary disorders, including lung cancer, hilar lymphadenopathy, metastatic pulmonary nodules as small as 3 mm, pulmonary hemorrhage, pulmonary edema and bronchial wall thickening in bronchiectasis. CONCLUSION: Our preliminary results indicate that the HASTE sequence provides a practical means for breath-hold MR imaging of lung parenchyma. PMID- 10374664 TI - An attempt of pulmonary perfusion imaging utilizing ultrashort echo time turbo FLASH sequence with signal targeting and alternating radio-frequency (STAR). PMID- 10374665 TI - Mediastinal and diffuse pulmonary haemangiolymphangioma. PMID- 10374666 TI - Quantitative assessment of angiogenesis in the chick embryo and its chorioallantoic membrane by computerised analysis of angiographic images. AB - We studied, in vivo, the angiogenesis process in the chick embryo and its chorioallantoic membrane (CAM) using digital subtraction angiography (DSA) in conjunction with computer-assisted image analysis. In a series of fertilised eggs, angiography was carried out at days 8, 10, 12 and 14 of embryonic development. The angiographic images were digitised and subsequently processed for a specific image analysis. A set of specific morphological parameters has been defined to allow an analytical characterisation of the vascularity status. Vessels were classified into three categories according to their diameter (50 100, 100-200, and > 200 microm). The data were normalised and statistically evaluated. Graphs showing the development of angiogenesis were obtained. Total vascular area revealed a continuous rise, whereas, total vascular length increased until day 12 and then it started decreasing. These morphometric parameters in the first two vessel categories progressively increased throughout the entire period of development, whereas in the third category they increased until day 10 and then they started decreasing. By applying a vascular casting technique CAM vessels were visualised and compared with those extracted from the processed angiographic image. The comparison revealed that there is exact matching for the first two vessel categories (diameters higher than 100 microm) while the matching of the third category (diameters between 50 and 100 microm) is approximate. PMID- 10374667 TI - Angiostatic treatment of neuroblastoma. AB - Neuroblastoma is a malignant solid tumor of childhood with a poor prognosis. The growth of solid tumors has been shown to be dependent on new blood vessel formation, i.e. angiogenesis. Several steps in the metastatic process have also been found to be angiogenesis-dependent. Neuroblastomas grow quickly, are highly vascularized, and metastasize early, and hence inhibition of angiogenesis- angiostatic therapy--may be indicated in this disease. In order to investigate the effects of angiostatic agents in this disease, a new animal experimental model for human neuroblastoma was developed. Three angiostatic agents were tested in the model: TNP-470, the synthetic analogue of fumagillin, given subcutaneously, and the endogenous steroid 2-methoxyestradiol and its derivative 2-propynylestradiol, given orally. TNP-470 administration resulted in a significant reduction of the tumor growth rate and microvascular counts, and of the fraction of viable tumor cells, compared to controls. The fraction of apoptotic tumor cells increased threefold, while that of proliferative cells remained unaltered. This can explain the reduced net growth. Treatment with the angiostatic and chemotherapeutic steroids 2-methoxyestradiol and 2 propynylestradiol yielded similar results. However, the mechanism of action of these steroids was bimodal; the effect occurring both through inhibition of tumor angiogenesis and through induction of tumor cell apoptosis. It was shown for the first time that inhibition of angiogenesis regardless of agent induces striking chromaffin differentiation, observed as increased expression of insulin-like growth factor II gene, tyrosine hydroxylase, and chromogranin A, and increased formation of cellular processes. It is suggested that inhibition of angiogenesis induces metabolic stress, resulting in chromaffin differentiation and apoptosis. Such agonal differentiation may be the link between angiostatic therapy and tumor cell apoptosis. Angiostatic agents administered as single therapy have an objective tumoristatic effect in our neuroblastoma model. Angiostatic treatment of neuroblastoma is a new and promising treatment modality that merits clinical investigation. PMID- 10374669 TI - A model for combined assessment of motor performance and behaviour in 3-year-old children. AB - This paper presents a new model for combined assessment of motor performance and behaviour (CAMPB) in 3-year-old children. It is intended for simultaneous use with a scale for assessment of motor-perceptual development. The child's performance is observed and compared with detailed descriptions of performance in gross and fine motor functions, and descriptions of coordination, attention and social behaviour, included in a protocol. An overall evaluation is also made. These assessments have been performed in a longitudinal follow-up study of children who needed intensive care neonatally and a control group of 72 neonatally healthy children. In this report the results from CAMPB assessments in the control group are presented. CAMPB together with the motor-perceptual scale was feasible in these 3-year-old children and CAMPB was sensitive enough to detect differences between children. The motor performance in most children conformed with the descriptions of gross and fine motor function in the protocol, and clear deviations were few. Seven per cent of the children had considerable problems in motor function and/or perception, in combination with a lack of attention, according to the overall evaluation. PMID- 10374668 TI - Short-term outcome of perinatal care in a Swedish county. Mortality, neonatal intensive care and overall evaluation of neuromotor function at 0-10 months of corrected age in preterm and term infants. AB - Improvements in obstetrical and neonatal care during the last decades have led to a marked increase in survival rate of preterm and term infants. In order to study the short- and long-term outcome in infants who survived neonatal intensive care (NIC) and were born in the county of Uppsala between January 1st 1986 and April 30th 1989, a prospective long-term follow-up study was conducted. Epidemiological data on all infants born in the county during the study period and the short-term outcome, measured as overall neuromotor function at term and at 2, 4, 6 and 10 months of corrected age in 245 infants surviving NIC and 72 healthy control infants are presented. The infants' neuromotor function was evaluated with different clinical neurological methods. In the study population of NIC infants 85.9% survived the neonatal period. The early infant mortality was high in this group 11.6% compared to that of all infants born in the county of Uppsala (0.30%). Only a minority of the infants showed abnormal neuromotor function. A comparison of the results of the overall evaluation of neuromotor function at 10 months of age with those of the examinations made at an earlier age showed poor correspondence in individual infants, especially in preterm and very preterm infants. PMID- 10374670 TI - A classification of problems regarding gut endocrinomas (carcinoids and relevant neoplasms). AB - In the field of gut endocrinomas (carcinoids and relevant neoplasms), several classifications have been internationally accepted and utilized at varying frequency. The basis of the concepts regarding gut endocrinomas from which these classifications have been proposed were drawn from the different aspects. These included embryology, histologic growth patterns, histochemistry including silver impregnations, electron microscopic morphology of endocrine secretory granules, endocrine cell types and histologic morphology along with functional characteristics, supplemented by immunohistochemical features. Due to continuous progress being made in this particular field of research and the many new discoveries made by pioneering investigators, the concepts of gut endocrinomas have been modified and revised during such long history of the research activities. This study aims to re-evaluate these classifications in relation to the concepts of gut endocrinomas, and to select and supply rearranged classifications that may be easily utilized for practical purposes. This study also proposes a comprehensive overview of histogenesis in the gut endocrinoma group. This consists of typical carcinoids and their atypical variants taken in relation to the carcinomatous group including ordinary carcinomas and their variants with endocrine elements. Special emphasis is given to the point that there is a gradual transition, one without a definite boundary between these tumors. PMID- 10374671 TI - Somatostatinoma/inhibitory syndrome: a statistical evaluation of 173 reported cases as compared to other pancreatic endocrinomas. AB - Somatostatin is known to inhibit the secretory release of other peptide hormones. Somatostatinomas associated either with or without somatostatinoma (inhibitory) syndrome are rare neoplasms among gut-pancreatic endocrinomas. Collected from international literature, this study aimed to perform a statistical analysis on 173 patients with somatostatinoma/inhibitory syndrome. The evaluation further attempted to provide investigators in this particular field of research with extensive and precise information on the present situation of somatostatinoma. The 173 patients consisted of 81 with pancreatic somatostatinomas and 92 with extrapancreatic somatostatinomas. Most of the latter were found to have originated in the duodenum and may be termed as carcinoid somatostatinoma. Where data were considered to be adequate, a comparative study was carried out between two groups, pancreatic and duodenal, each consisting of 81 patients. A statistically significant difference between these two groups was found in the incidence of inhibitory syndrome (18.5% versus 2.5%) and von Recklinghausen's disease (1.2% versus 43.2%), large size of tumor (>20 mm) (85.5% versus 41.4%), multisecretory activities (33.3% versus 16.3%), and presence of psammoma bodies (2.5% versus 49.4%). There was no statistically significant difference in the rate of metastases and malignancy between the two groups. The average postoperative 5-year survival rate was 75.2% in 90 patients overall, 59.9% in 44 with metastases and 100.0% in 46 without metastases. Compared with the other pancreatic endocrinomas, including PPomas, glucagonomas, vipomas, gastrinomas, and insulinomas, somatostatinomas were characterized by the low rate of the relevant syndrome and multiple endocrine neoplasia syndrome type 1. There was a low rate of multiplicity, and a high incidence of psammoma bodies in the duodenal group particularly with von Recklinghausen's disease. A high rate of malignancy was recorded, resulting in a low postoperative survival rate of patients with metastases. In conclusion, somatostatinomas exhibited characteristic features quite different from those of the other pancreatic endocrinomas regarding multiple points. PMID- 10374672 TI - Significance of lymph node metastases in the surgical management of pancreatic head carcinoma. AB - Recent reports have demonstrated a reduction in the morbidity and mortality of pancreatic resections and improvement in the 5-actuarial survival for patients with resected ductal adenocarcinoma. However, the prognosis for patients with lymph node metastases remains uncertain. The purpose of this study is to determine if the presence of lymph node metastases influences the survival in patients with otherwise potentially curable pancreatic head carcinoma. Between January 1974 and December 1995, 340 patients with pancreatic carcinoma, including 238 patients with pancreatic head tumours, were evaluated and treated in our Department. Seventy-seven (32.3%) patients with pancreatic head carcinoma underwent pancreaticoduodenectomy. Ages ranged from 40 to 76 years, with a mean age of 61 years. Fifty patients were male, twenty-seven were female. The overall postoperative mortality rate was 5.2% (4 patients) and morbidity was 23.4%. Median survival following resection was 17 months (range 0 to 79). The estimated 1-, 2-, 3- and 5-year survival were 68.8%, 48.1%, 23.4% and 18.2%, respectively. There were 14 five-year survivors. Of the 77 patients, 25 (32.5%) had negative lymph nodes. The median and 5-year survival in these node-negative patients were 33 months (range 5 to 79) and 40%, respectively. Whereas the median survival and 5-year survival in 52 patients with lymph nodes metastases were 14 months (range 0 to 61) and 7.7%, respectively (P<0.0001). There were 4 five-year survivors in the group of patients with lymph node metastases; in 2 patients was performed extensive lymph node dissection (R2) and in other 2 patients R1 procedure. In the patients with lymph node metastases undergoing R1 resection (n = 39), the 1-, 2- and 5-year survival rates were 48.7%, 23.1% and 5.1%, respectively. Whereas in the patients with positive lymph nodes undergoing R2 resection (n = 14), the 1-, 2- and 5-year survival rates were 92.9%, 64.3% and 14.3%, respectively (P<0.02). As expected, tumour size and margin status in specimen proved to be two significant factors predicting survival. Pancreatoduodenectomy can be performed with low operative mortality. Lymph nodes metastases are found in 67.5% of patient undergoing resection. Pancreaticoduodenectomy offers good palliation for patients with lymph nodes metastases and encouraging long-term survival rates as well as a chance for cure in patients with negative lymph nodes and negative margins of resection. PMID- 10374673 TI - Colorectal cancer complicating ulcerative colitis: an institutional series. AB - Ulcerative colitis predisposes to colorectal cancer: the risk increases along with disease duration and extension. Also some subsets of patients are at increased risk, namely patients with early onset of colitis, and patients with primary sclerosing cholangitis. Cancer complicating ulcerative colitis affects evenly all the colon, and is not located more frequently in the rectum and in the sigmoid colon, as well as the sporadic counterpart. Multiple cancers and cancers associated with high grade dysplasia are not infrequent in ulcerative colitis; for this reason, and for controlling the colitis, the treatment of choice is total colectomy, with or without colostomy. The prognosis of cancer complicating ulcerative colitis is similar to the sporadic counterpart. The Authors present a colon cancers series as a complication of colitis occurred at Regina Elena Cancer Institute of Rome, Italy, over the period 1975-1998. PMID- 10374674 TI - Deficient DNA repair capacity: a predisposing factor and high risk predictive marker in familial colorectal cancer. AB - Even though colorectal cancer tends to aggregate in families, there is paucity of information on the genetic determinism for familial colorectal cancer (FCRC) predisposition. Therefore, we investigated constitutional chromosome abnormalities and bleomycin induced chromosome sensitivity of 26 familial and 30 sporadic colorectal cancer (SCRC) patients, 60 unaffected family members (first/second degree relatives) and 30 normal healthy controls to determine whether these parameters could give any clues on genetic predisposing factors by which high risk members in CRC families could be identified. The test assay used bleomycin-induced chromatid breaks in short term microcultures of peripheral blood lymphocytes of the subjects. The CRC patients, the unaffected family members and the controls did not show any constitutional chromosomal abnormalities. However, with regard to bleomycin sensitivity, there was significant difference between the CRC patients, unaffected relatives and controls. The mean b/c values of 1.64+/-0.42 for the FCRC patients and 1.08+/ 0.34 for the SCRC patients were significantly higher than the mean b/c values of 0.62+/-0.18 for the unaffected relatives and 0.52+/-0.12 for the controls (P<0.001). A noteworthy observation was that 6 unaffected members from 6 CRC families also showed bleomycin hypersensitivity, at the initiation of the study. Since they expressed mean b/c values greater than 1.0, which was as high a value as those of the patients, they were regularly followed up. Out of these 6 members, 2 developed CRC later. This clearly demonstrates that mutagen hypersensitivity among unaffected relatives in CRC families may be related to cancer predisposition. Hence, this cytogenetic assay could be utilised to identify the genetically high risk individuals in CRC families. PMID- 10374675 TI - Differential effect of protein-bound polysaccharide (PSK) on survival of experimental murine tumors. AB - The effect of protein-bound polysaccharide (PSK) on the survival of BALB/c and C57BL/6 mice after intravenous injections of syngeneic murine sarcomas (GR9.B9 and Meth-A), LSTRA lymphoma and B16 melanoma cells was studied. Pretreatment of mice with PSK significantly increased survival after the injection of either type of sarcoma cells, although the effect was attenuated when high numbers of cells were injected. Survival was not modified significantly in LSTRA lymphoma or B16 melanoma. Mice pretreated with anti-asialo GMI serum showed significantly decreased survival from all tumors in comparison with untreated mice injected with tumors, regardless of cell dose used. We observed an inverse correlation between H-2 antigen expression and in vitro NK sensitivity of tumor cells from all lines except B16 melanoma cells. These results clearly suggest that pretreatment of mice with PSK prolongs survival and inhibits metastasis formation in mice injected with sarcoma cells, being this effect highly selective, since survival was not improved in mice injected with LSTRA lymphoma or B16 melanoma. PMID- 10374676 TI - Human papillomavirus infection and p53 nuclear overexpression in anal canal carcinoma. AB - The product of HPV E6 and E7 genes is able to inactivate both the p53 and pRb proteins. The aim of this study was to evaluate the correlation among anal HPV infection and nuclear p53 overexpression. The Authors evaluated HPV DNA by PCR and p53 nuclear expression by immunohistochemistry in 12 cloacogenic and 6 squamocellular carcinoma. HPV DNA was detected in 71.4% of the squamocellular tumors and in 57.1% of the cloacogenic tumors. In squamocellular tumors HPV types 31-33 and 16 were found; in cloacogenic tumors type 16 alone was detected. Nuclear accumulation of p53 was found to be associated with the presence of HPV. There was no significant difference in parietal infiltration, lymph nodes involvement and prognosis between HPV+p53+ patients and HPV-p53- patients. Tumor aggressiveness is likely to be enhanced by factors other than HPV infection and p53 overexpression. PMID- 10374677 TI - Multiple primary cancers and HPV infection: are they related? AB - Multiple primary cancers have been reported with increasing frequency in recent years, but the presence of foreign DNA sequences of infectious agents in tumours arising in the same patient has so far not been investigated. We report a case of a patient with Hodgkin's lymphoma, an "in situ" cervix carcinoma and an adenocarcinoma of the right and left mammary gland. In all the tumour samples we detected the presence of DNA genomic sequences of Papillomavirus type 16. Our results suggest that HPV infection may be an exogenous risk factor even in second primary tumours of non-epithelial origin. PMID- 10374679 TI - Hypercalcemia in patients with esophageal cancer. AB - Hypercalcemia is a paraneoplastic syndrome that is associated with squamous cell cancers and which may be of life-threatening proportions. We investigated the incidence and prognostic importance of hypercalcemia in patients with esophageal cancer at the Department of Veterans Affairs Medical Center, Washington, DC, USA. The medical records of 170 patients with esophageal cancer from January 1988 to January 1998 were examined. Of the 170 patients with esophageal cancer, 47 (27.6%) had hypercalcemia during the course of their disease. Five (10.6%) of the 47 hypercalcemic patients were found to have hypercalcemia at the time of diagnosis. Forty-six of the 47 hypercalcemic patients had squamous cell carcinoma and 1 had adenosquamous cell carcinoma. Seven (14.8%) had bony metastasis. The median survival of patients with hypercalcemia and esophageal cancer was 12.4 months and 12.6 months for patients without hypercalcemia. Hypercalcemia is a common complication of squamous cell esophageal carcinoma. The survival of patients with or without this complication is similar; thus, it may not be a poor prognostic factor. PMID- 10374678 TI - GM-CSF, ARA-C, VP-16 and idarubicin (GM-IVA), a short, and effective induction treatment for de novo AML, suitable for the elderly. AB - GM-IVA is a short and effective induction therapy of non M3 de novo AML including GM-CSF (300 mcg 12 hrs before starting therapy), Ara-C (250 mg/sqm c.i. x 3 days), VP16 (100 mg/sqm x 3 days) and idarubicin (12 mg/sqm x 3 days); it was followed by a fludarabine containing salvage protocol (FLANG). Patients <60 years of age achieving CR received 2 courses of FLANG and autologous or allogeneic BMT when possible. Patients >60 years of age in CR received a second course of GM IVA. Twenty-one consecutive patients (mean age 64, range 29-85) entered the study. Three patients (14%) died during induction therapy. After one course of GM IVA, CR was achieved in 12 patients (57%). Two further patients were salvaged with FLANG therapy so that the final CR rate was 14/21 (67%). In elderly patients the final CR rate (62%) is noteworthy, considering that 6 patients were >70 years of age and 3 were >80. All three patients >80 achieved CR (lasting 5 to 7 months). The median time of granulocyte and platelet recovery was 15 days. Our scheme was well tolerated. In the group of elderly patients 3 out of 14 died during induction (21%) and 4 life-threatening infections were observed (28%). The short duration of cytotoxic therapy and perhaps the use of G-CSF contributed to a reduction of the hospitalization period (median of 22 days), thus providing major savings on induction costs and allowing for better utilization of beds as well as significantly improving patients' quality of life. PMID- 10374680 TI - Endoscopic palliative therapy in neoplastic diseases of the esophagus. AB - Over the past 20 years, fiberoptic endoscopy, followed by video-endoscopy, has taken on an increasingly important role in the diagnostics and therapy for digestive tract diseases. Especially in the field of oncology, endoscopy is fundamental not only for the diagnosis and staging of diseases of the gastroenteric tract but also as definitive and/or palliative therapy for tumors that do not respond to radical treatment. Endoscopic techniques are widely employed in diseases of the esophagus and the head and neck district. In fact, it is generally accepted that only 35% of patients with cancer of the esophagus or of the cardias may benefit from surgery with 5-year survival rates of about 5% in Western countries (1, 2). PMID- 10374681 TI - Sentinel node biopsy and selective lymph node dissection in cutaneous melanoma patients. AB - Sentinel node biopsy allows an accurate selection of melanoma patients to be submitted to therapeutic dissection. From February 1994 to August 1998, at the National Cancer Institute, S. Pio X Hospital in Milan and Bufalini Hospital in Cesena, 580 sentinel node biopsies were performed in 540 stage I melanoma patients (242 males; 298 females; median age 47). Primary melanoma was located in the trunk in 201 patients, in lower limbs in 242 cases, in upper limbs in 80 cases and in head and neck in 17 patients. Injection of blue dye for sentinel node identification was performed in all cases; 372 patients were submitted to preoperative lymphoscintigraphy and in 272 cases an intraoperatory probe for a radioguided biopsy was utilized. Sentinel node identification rate was 91%. Sentinel node positivity rate was 15%. Frozen sections were examined in 199 cases. Distribution of positive cases according to primary thickness is the following: <1 mm: 1%; 1-1.99 mm: 5%; 2-2.99 mm: 18% and > or =3 mm: 27%. Sentinel node appeared to be the only metastatic node in 77% of patients submitted to dissection. The adoption of preoperative lymphoscintigraphy and the intraoperative use of the gamma probe contributed substantially in S.N. identification. No complications caused by the procedure were reported. Eight patients had a regional node relapse after a negative sentinel node biopsy and were submitted to therapeutic distant dissection. Currently 513 patients are alive with no evidence of disease. Present data confirm the feasibility and safety of sentinel node technique for selection of patients to be submitted to radical node dissection and to eventual adjuvant treatments. PMID- 10374682 TI - Lymphatic mapping for the dissection of the sentinel node in the treatment of breast carcinoma. AB - The Authors present the preliminary results of a study aimed at verifying the effectiveness of lymphatic mapping for the dissection of the sentinel lymph node in the treatment of breast carcinoma. The lymphoscintigraphy method was used in the study performed on 24 patients to evidence the sentinel lymph node. The sentinel lymph node was identified in 23/24 patients (95,8%), with 100% accuracy, negative predictions were 100% (21/21). Consistently with the low number of cases treated, our results seem to guarantee the same prognostic accuracy obtained using radical lymphadenectomy extended to the third lymph node level. PMID- 10374683 TI - Lymphnode metastasis in head and neck squamous cells carcinoma: multivariate analysis of prognostic variables. AB - Cervical lymphnodes metastatization by the squamous cell carcinoma of the head and neck is well known as a prognostic negative factor as far as survival is concerned. Multivariate analysis has been used on 207 cases of head and neck squamous cell carcinomas (HNSCC) in order to identify the possible prognostic significance of a group of clinical and histopathological characteristics, aiming to find a correlation with the possible occurrence of cervical lymphnodes. Two hundred and seven patients (168 males and 39 females, mean age: 62 years) with SCCHN were studied. They underwent surgery alone and radiotherapeutic associated treatment. Variables regarding the patient, carcinoma and histology were analysed: age, sex, smoking and alcohol consumption, performance status, concomitant internal pathologies (cardiopathies, hepatopathies, broncho pneumopathies, metabolic disorders), site and size of primary tumor (T stage), number and size of laterocervical lymph node localization (clinical N stage), grading, vascular permeation, perineural infiltration. Multivariate analysis of prognostic factors was performed using BMDP's PLR programme. Some variables showed a great risk of lymphnode metastasis; among sites: supraglottic larynx (p = 0.05), base of the tongue (p = 0.04), hypopharynx (p = 0.05); some histological parameters as lower degree of histological differentiation (p = 0.02), the presence of vascular permeation (p = 0.06) and perineural invasion (p = 0.07) appear to represent predisposing factors for the onset of adenopathies. By considering prognostic factors as shown, it is possible to better identify metastasis risk cases, that leads to improved therapeutical strategies. PMID- 10374684 TI - Vitiligo, autoimmune thyroiditis: a rare thyroid cancer arising with bone metastates on maxillofacial area. AB - An association between vitiligo and autoimmune thyroid disorders had previously been postulated. Thyroid disorders were found in 18.5% of 15,126 patients with vitiligo, on the basis of the anamnestic data. Then, we investigated 255 healthy relatives in whom we tested only T3, T4 and TSH. With the immunological investigation we detected a higher incidence of TMA in vitiligo patients and in the family members. Therefore, on the basis of the immunologic and thyroid pathology functional data, we observed a thyroid pathology in 25% of the 890 vitiligo patients and in 21.1% of their first degree relatives. Then, clinical observation enabled to discover that 3 of 15,126 patients had undergone exeresis for a thyroid carcinoma and in the 890 vitiligo patients, who had undergone particular investigations, we found a thyroid carcinoma in 3 subjects. In one case lymphnodal involvement and bone metastases in the maxillary district were found. The purpose of this work is to evaluate the incidence of thyrosis and of thyroid carcinoma in vitiligo patients observed for 20 years. PMID- 10374685 TI - Osteosarcoma arising in a mature cystic teratoma of the ovary. AB - A 80-year old woman was found to have an osteosarcoma arising within a mature cystic teratoma of the ovary. To the Author's knowledge, this is the fourth reported case of osteosarcoma arising within a teratoma. PMID- 10374686 TI - Isolated splenic metastasis from endometrial carcinoma. AB - Splenic involvement by carcinomatous metastases occurs late in the course of widely disseminated disease. Solitary metastasis to the spleen from a primary of any source is an uncommon event. We report a case of a 58-year-old woman in which an asymptomatic solitary metastatic endometrial adenocarcinoma of the spleen was detected 28 months after resection of the primary malignancy and treated by splenectomy. There are thirteen other reported cases of solitary splenic metastasis from gynecological cancers; in four of these reports the source was endometrial. PMID- 10374687 TI - Allele frequency of D1S191 microsatellite locus in Japanese people. AB - The allele frequency of D1S191 microsatellite locus was analyzed in 398 normal tissues of Japanese adults. The frequency of D1S191 microsatellite polymorphism was 90.2% (359/398). The size of D1S191 PCR fragments ranged from 147 bp to 169 bp. The most frequent allele in the Japanese subjects was 161 bp (34.1%) followed by 159 bp (27.9%), 163 bp (17.7%), and 157 bp (12.4%) fragments. The observed allelic distribution of this microsatellite in the Japanese subjects was similar to that of European Caucasians deposited in the Genome Database. PMID- 10374688 TI - Frequent Bax frameshift mutations in gastric cancer with high but not low microsatellite instability. AB - Widespread microsatellite instability (MSI) due to the defective DNA mismatch repair underlies the pathogenesis of the majority of hereditary non-polyposis colorectal cancer and a subset of various sporadic malignant tumors. Using 5 microsatellite markers and the criteria of MSI proposed by the National Cancer Institute (NCI) workshop, we analyzed 205 gastric adenocarcinomas for MSI. Based on the number of markers showing instability per tumor, the tumors were divided into three groups; those with two or more of the five markers displaying instability (high MSI, MSI-H), those with one of five markers displaying instability (low MSI, MSI-L), and those with no instability (microsatellite stable, MSS). Among 205 tumors, 30 (15%) were MSI-H, 15 (7%) were MSI-L, and 160 (78%) were MSS. All of the 30 MSI-H tumors demonstrated instability at BAT26, a sensitive marker for the widespread MSI, while none of the 15 MSI-L tumors did. MSI-H tumors were significantly associated with distal location and well or moderate differentiation, but MSI-L tumors were indistinguishable from MSS tumors. Bax frameshift mutations were detected in 60% of the 30 MSI-H tumors, while not in any of the 15 MSI-L tumors. These results suggest that microsatellite analysis using the criteria proposed by the NCI workshop may be appropriate for gastric cancers because it unveils real differences in genotype and phenotype. PMID- 10374689 TI - Basal expression and cytokine induction of intercellular adhesion molecule-1 in human pancreatic cancer cell lines. AB - Although expression of intercellular adhesion molecule-1 (ICAM-1) expression has been demonstrated in many human malignancies, very little is known about the ICAM 1 expression in human pancreatic cancer. We have examined ICAM-1 expression in pancreatic cancer cell lines and the induction of ICAM-1 in these cells by flow cytometry. The degree of baseline ICAM-1 expression was most significant in Panc 1, followed by PMH-2, Capan-2, Bx-PC-3, Capan-1, and PMH-3 in that order. PaCa-2 and AsPC-1 showed very low levels of baseline ICAM-1 expression. After tumor necrosis factor alpha and interferon gamma stimulation, five cell lines exhibited distinct ICAM-1 induction. The degree of induction was remarkable in AsPC-1 (32 fold), and moderate in PMH-3 (6.5-fold), PaCa-2 (3.2-fold), Capan-1 (1.6-fold), and BxPC-3 (1.5-fold). The ICAM-1 expression levels of PMH-2 and Capan-2 after stimulation were nearly the same as those before stimulation (1.2-fold and 1.1 fold, respectively). These results suggest that ICAM-1 is overexpressed and inducible by tumor necrosis factor alpha and interferon gamma in pancreatic cancer cell lines. PMID- 10374690 TI - Carcinogenesis of intestinal-type gastric cancer and colorectal cancer is commonly accompanied by expression of brain (fetal)-type glycogen phosphorylase. AB - Our previous studies have demonstrated the significant enzymatic activity of glycogen phosphorylase (GP) in the gastric carcinoma and proliferating cells of particular intestinal metaplasia (IM). This paper reviewed the identification of the GP isoform in the gastrointestinal carcinoma, and the investigation on the role of this molecule in the gastrointestinal carcinogenesis. The only isoform expressed in gastric cancer was brain-type GP (BGP) using polymerase chain reaction (PCR) analysis. The expression of BGP, oncogene products and proliferating cell nuclear antigen in the gastric and colorectal carcinomas, their premalignant lesions, and the normal mucosa were examined using 136 gastric and 96 colorectal surgically resected specimens, and 55 endoscopically resected colorectal adenomas. The BGP visualized by immunohistochemistry was commonly present in intestinal-type gastric (80.6%) and colorectal (83.3%) carcinomas, whereas no BGP expression was seen in the normal human gastric and large intestinal mucosa except in the BGP foci described below. IMs with BGP had close correlation with intestinal-type gastric carcinoma, and some of them coexpressed accumulated p53 protein. The expression of BGP during 'adenoma carcinoma sequence' (ACS) showed excellent correlation with the increased dysplasia and was found prior to p53 expression. Positive staining in overtly normal looking colonic mucosa (BGP foci) was observed mainly around carcinomas without any adenoma component, and frequent p53 mutation (41.2%) was detected in the BGP foci using PCR-single strand conformation polymorphism analysis. It is suggested that BGP is a novel biomarker for carcinogenesis in the intestinal-type gastric carcinoma and in both of the pathways of ACS and the 'de novo' colorectal carcinoma. PMID- 10374691 TI - Semi-quantitative procedure for telomeric repeat amplification protocol (TRAP) assay in colorectal carcinomas. AB - Telomerase activity is intensively studied as a prognostic marker in malignancies, and generally detected by telomeric repeat amplification protocol (TRAP) assay. Recently, Ohyashiki et al. demonstrated a semi-quantitative procedure for TRAP assay, which is more useful than the qualitative one to analyze the correlation between telomerase activity and clinicopathological features in carcinomas. Therefore, we used semi-quantitative TRAP assay to analyze the correlation between telomerase activity and clinicopathological factors in colorectal carcinomas. Twenty-nine cases of advanced colorectal carcinoma, obtained by surgery, were studied. A telomerase detection kit, TRAP eze, was used for the PCR based TRAP assay, and a DNA sequencer for a semi quantitative analysis was used for a analysis. Telomerase activity was positively detected in all samples, and its mean value was 2.076+/-1.634 units (range; 0.536 8.932 units). Based on Dukes' classification, the activity tended to be higher in Dukes' A (2.722 units; P = 0.0525) and C (2.430 units) than in Dukes' B (1.552 units). The activity was higher in the right colon than at other locations. (right vs. left, rectum = 3.167 vs. 1.216 units; P<0.05, 1.604 units; P<0.01). The activity was higher in poorly differentiated adenocarcinoma than in well differentiated adenocarcinoma (3.743 vs. 1.491 units; P<0.05). Semi-quantitative TRAP assay is useful to analyze the correlation between telomerase activity and clinicopathological factors in colorectal carcinomas. PMID- 10374693 TI - Human CD4+ T cell differentiation and effector function: implications for autoimmunity. AB - Human CD4+ memory T cells progress through stages of postthymic differentiation that have been characterized by distinct phenotypes. We have investigated the factors regulating cytokine production, and the correlation between phenotype and effector function in normal and autoimmune individuals. These studies suggest that antigen-induced proliferation in the periphery drives CD4+ T cells through successive stages of differentiation that culminate in optimal effector function and resistance to external modulatory influences. Moreover, these studies support the concept that in autoimmune individuals, the chronic accumulation of differentiated proinflammatory T cells perpetuate the inflammatory response resulting in aggressive disease. PMID- 10374694 TI - The influence of base sequence on the immunostimulatory properties of DNA. AB - DNA is a complex macromolecule whose immunological properties vary with base sequences. As shown with synthetic oligonucleotides, potent immune stimulation results from six base motifs called CpG motifs or immunostimulatory sequences (ISS). These sequences center on an unmethylated CpG dinucleotide and occur much more commonly in bacterial DNA than mammalian DNA. As such, CpG motifs may function as a danger signal to stimulate B cell activation and cytokine production. In addition to CpG motifs, runs of deoxyguanosine (dG) residues in DNA can induce B cell activation and promote macrophage cytokine expression by adjacent CpG motifs. The array of these sequences may determine the overall immune activity of a DNA molecule and affect such processes as host defense against infection as well as the use of plasmids and synthetic oligonucleotides to treat disease. PMID- 10374692 TI - CD28/CTLA-4 and CD80/CD86 families: signaling and function. AB - T cell stimulation in the absence of a second, costimulatory signal can lead to anergy or the induction of cell death. CD28 is a major T cell costimulatory receptor, the coengagement of which can prevent anergy and cell death. The CD28 receptor is a member of a complex family of polypeptides that includes at least two receptors and two ligands. Cytotoxic lymphocyte-associated molecule-4 (CTLA 4, CD152) is the second member of the CD28 receptor family. The ligands or counterreceptors for these two proteins are the B7 family members, CD80 (B7-1) and CD86 (B7-2). This article reviews the CD28/CTLA4 and CD80/CD86 families, and outlines the functional outcomes and biochemical signaling pathways recruited after CD28 ligation. PMID- 10374698 TI - T cells in pregnancy: illusion and reality. PMID- 10374699 TI - The role of gamma/delta T cell receptor positive cells in pregnancy. AB - PROBLEM: Due to the lack of classical HLA antigens on the trophoblast, fetal antigens are possibly presented in a non major histocompatibility complex (MHC) restricted way. Decidual gammadelta T cells, which significantly increase in number during pregnancy, might play a role in recognition of fetal antigens and also in determining the quality of the response to these antigens. Our study was aimed at investigating the role of this cell population in progesterone-dependent immunomodulation. METHOD OF STUDY: Peripheral lymphocytes from healthy pregnant women and from habitual aborters were tested by immunocytochemistry for the presence of gamma/delta T cell receptor (TCR) and progesterone receptor. To investigate the effect of treatment with a pan anti gamma/delta antibody, lymphocytes were incubated for 3 hr with the antibody, and then interleukin (IL) 10, IL-12 and progesterone-induced blocking factor (PIBF) expression (by immuno cytochemistry) as well as natural killer (NK) cell activity were determined. RESULTS: In peripheral blood of healthy pregnant women the percentage of gamma/delta TCR+ cells was significantly higher (P < 0.001) than in that of recurrent aborters or of non-pregnant individuals. Ninety-seven percent of gamma/delta TCR+ pregnancy lymphocytes expressed progesterone receptor. Binding of a specific antibody to the gamma/delta TCR inhibited PIBF- as well as IL-10 production, whereas it increased NK activity and IL-12 expression. CONCLUSIONS: These data suggest the role of gamma/delta TCR-bearing lymphocytes in progesterone-dependent immunomodulation. PMID- 10374697 TI - Vaccine strategies to prevent rheumatic fever. AB - Group A streptococci (GAS) are responsible for numerous human illnesses, ranging from pharyngitis to severe invasive infections, such as necrotizing fascitis and toxic shock syndrome to the postinfectious sequelae, acute rheumatic fever (ARF), and glomerulonephritis. To date, to develop a vaccine, studies have focused on the M protein. However, designing a vaccine to prevent GAS infection based on this molecule has been hampered by the vast number of M protein serotypes and the possibility that it may induce potentially harmful autoimmune reactions. In this article, the authors discuss recent approaches to overcoming the problems of an M protein-based vaccine. In addition, recent studies identifying the protective properties of other streptococcal antigens and their potential as vaccine candidates are discussed. PMID- 10374696 TI - Immunological self/nonself discrimination: integration of self vs nonself during cognate T cell interactions with antigen-presenting cells. AB - The hypothesis is presented that immunological integration of nonefficacious vs efficacious T cell antigen receptor (TCR) signals are foundational for self/nonself discrimination and that multiple integrative mechanisms are intrinsic to the molecular to molar organization of an adaptive immune response. These integrative mechanisms are proposed to adaptively regulate expression of costimulatory signals, such that foreign proteins are associated with the expression of costimulatory signals, whereas self-proteins are associated with the lack of costimulatory signaling. Overall, this model offers several unique contributions to the study of immunology. First, this model postulates that cognate TCR/major histocompatibility complex (MHC) interactions are sufficient to adaptively mediate immunological self/nonself discrimination. This model thereby offers a unique alternative to models that largely rely on innate immunity to prime immune discrimination. Second, the integrative model argues that the immune system can simultaneously reinforce self-tolerance and promote immunity to foreign organisms at the same time and in the same location. Many alternative models presume that pathogenic self-reactive T cells do not exist at the outset of an immune response against foreign agents. Third, the integrative model uniquely predicts relationships between immunodeficiency and autoimmune pathogenesis. Fourth, this model illustrates the regulatory advantages of cognate antigen presenting cell (APC) systems (i.e., T cell or B cell APC) compared to nonspecific APC. Cognate APC systems together with the respective clonotypic responders may comprise a fundamental "network" of lymphoid cells. Such networks would have clone-specific regulatory capabilities and may be central for immunological self/nonself discrimination. Fifth, this model provides an explanation for "infectious" tolerance without creating specialized subsets of "suppressor" or "regulatory" T cells. Each mature T cell retains the potential to reinforce tolerance or mediate immunity, depending on the specific antigenic cues present in the immediate environment. PMID- 10374701 TI - The level of human decidua-associated protein (hDP) 200, identified as a monoclonal rheumatoid factor in the fallopian tube. AB - PROBLEM: The aim of this study was to examine whether ectopic pregnancy is associated with increased levels of human decidua-associated protein (hDP) 200, identified as a monoclonal rheumatoid factor, in the related fallopian tube as compared with normal tubes. METHOD OF STUDY: Tubal fluid samples were obtained prospectively from two groups of patients: group I, 20 patients admitted for total abdominal hysterectomy and bilateral salpingo-oophorectomy (TAH and BSO); group II, 14 women with ectopic pregnancy, undergoing unilateral salpingectomy by laparoscopy or laparotomy. hDP 200 was assayed using double-site enzyme-linked immunosorbent assay. RESULTS: Very low levels of hDP 200 were found in normal tubes, while tubes with ectopic pregnancy demonstrated significantly higher levels of tubal fluid hDP 200 (P < 0.00001). In addition, in women with ectopic pregnancy, the uterine fluid level of hDP 200 was significantly lower than the level of hDP 200 in the tube containing ectopic pregnancy (P < 0.0001). CONCLUSION: The increased level of hDP 200 (a monoclonal rheumatoid factor) in the tube containing ectopic pregnancy, as compared with uterine cavity and normal tubes, might be associated with the implantation site of the pregnancy. PMID- 10374700 TI - Human decidualized endometrial T lymphocytes do not substantially down-regulate CD3zeta. AB - PROBLEM: The T-cell antigen receptor (TCR) has been reported to be down-regulated on T-cells in the decidualized endometrium in early pregnancy. METHOD OF STUDY: The expression of CD3zeta, a component of the TCR complex, has been investigated in human first-trimester decidual T-cells using flow cytometric analysis of permeabilized cells. RESULTS: Levels of CD3zeta expression were significantly lower in decidual than in peripheral T-cells from non-pregnant women, as assessed by mean fluorescence intensity (4.2 vs. 5.5, logarithmic scale, P < 0.05). However, when decidual and peripheral T-cells from the same subjects were analyzed (n = 10), mean levels of CD3zeta were slightly, but not significantly, lower in decidual than in peripheral T-cells (P > 0.1). CD3zeta was not substantially down-regulated systemically as mean cytoplasmic CD3zeta levels did not differ significantly between peripheral blood T-cells from pregnant women and non-pregnant controls (P > 0.2). CD8+ cells outnumber CD4+ cells in decidua, but neither the proportions of these two T-cell subsets positive for cytoplasmic CD3zeta nor the mean levels of CD3zeta were significantly different. CONCLUSIONS: These results indicate that human decidual T-cells do not greatly down-regulate CD3zeta, but it is unclear if a small decrease in mean levels may be sufficient to compromise their capacity for activation. PMID- 10374695 TI - Negative signaling in health and disease. AB - Signal transduction induced by receptors can elicit intracellular biochemical events that either support or inhibit cell activation. Induction of the latter has been termed "negative signaling" and can be triggered by receptors on immune cells that are distinct from activating receptors while other growth-promoting receptors induce both positive and negative signaling events. Here, the biochemistry leading to cell activation or inhibition and induced by receptors on immune cells are reviewed. Furthermore, recent experimental evidence is reviewed that indicates an important contribution of negative signaling to the intracellular survival of infectious pathogens. PMID- 10374702 TI - Circulating interferon-gamma- and interleukin-4-secreting cells in recurrent spontaneous abortions. AB - PROBLEM: Are recurrent spontaneous abortions (RSAs) associated with deviation of circulating cytokine-secreting cells? METHOD OF STUDY: Interferon (IFN)-gamma- and interleukin (IL)-4 secreting cells were identified by enzyme-linked immunospot (ELISPOT) in blood from 34 women with RSA. Samples were taken before pregnancy and/or pregnancy weeks 7-10, 17-20, and after terminated pregnancy. Eleven healthy primigravidae and 10 non-pregnant women served as controls. RESULTS: No significant difference in numbers of IFN-gamma- and IL-4-secreting cells was noted within the RSA group when abortions and successful pregnancies were compared in samples taken before pregnancy. The number of IFN-gamma- as well as IL-4-secreting cells in pregnancy weeks 17-20 in the RSA group was significantly higher compared with before pregnancy, pregnancy weeks 7-10, and after pregnancy. In samples from non-pregnant women, the number of IFN-gamma- and IL-4-secreting cells was significantly higher in the RSA group compared with controls. CONCLUSIONS: Our results do not indicate a systemic shift in the general balance between T helper 1- and T helper 2-type cytokine pattern. A local shift at the fetomaternal interface seems more probable. PMID- 10374703 TI - Antineutrophil cytoplasmic antibodies and other immunologic abnormalities in patients with habitual abortion. AB - PROBLEM: The immunologic mechanisms of pregnancy loss in habitual aborters with antiphospholipid and antinuclear antibodies have not been fully clarified. The possible association of antineutrophil cytoplasmic antibodies (ANCAs) with recurrent miscarriage was examined. METHOD OF STUDY: In a prospective, controlled trial of 59 women with recurrent abortion, the prevalence of pANCA (antimyeloperoxidase), cANCA (antiproteinase-3), and immunoserologic abnormalities of systemic lupus erythematosus (SLE) anti double-stranded DNA, anti-SSA, anti-SSB, anti-U1RNP, anti-Sm, anticardiolipin and antinuclear antibodies, LE-cell, lupus anticoagulant, and complement-3 were investigated. RESULTS: pANCA occurred in 2, and cANCA in 6 of 59 case patients, but neither was observed in the controls (P = 0.09 for cANCA). cANCA levels were significantly higher in patients than in controls (P = 0.028). Six recurrent aborters were identified as having a group of immunoserologic abnormalities characteristic of SLE. CONCLUSIONS: Immunologic mechanisms detectable in SLE may operate in a subgroup of habitual aborters with suspected immunologic cause. ANCAs occur more frequently in patients with recurrent miscarriage than in controls. PMID- 10374704 TI - Heparin plus aspirin as a "single" therapy for recurrent spontaneous abortion associated with both allo- and autoimmunity. AB - PROBLEM: The aim of this study was to contribute to the study of the pathogenesis and the treatment of recurrent spontaneous abortion (RSA) associated with immune alterations. METHOD OF STUDY: This is a prospective clinical trial with 11 patients with RSA associated with allo- and autoimmunity not receiving lymphocyte immunizations but only heparin and aspirin preconceptionally and through pregnancy. A concurrent group of 8 patients receiving a complete therapy (lymphocyte immunizations, heparin, and aspirin) but not receiving heparin and aspirin preconceptionally is also included in this report. RESULTS: The rate of pregnancy success in these patients was 90.9% (10/11), and the rate of success of the concurrent group was 75.0% (6/8). CONCLUSIONS: The results are in agreement with the working hypothesis regarding the possible final common mechanism in the pathogenesis of abortion associated with allo- and autoimmunity. The "single" therapy with heparin and aspirin was effective, less costly, and logistically simpler to provide than a complete therapy including lymphocyte immunizations. PMID- 10374706 TI - Effect of serum on mouse granulated metrial gland cell cytotoxicity. AB - PROBLEM: To examine factors affecting mouse granulated metrial gland cell cytotoxicity. METHOD OF STUDY: Chromium-release assays using mouse metrial gland cell suspensions targeted against Wehi 164 fibrosarcoma cells were used. Modulation of cellular cytotoxicity was investigated using an antibody, anti asialo-GM1, to attempt depletion of effector cells and sera to determine if inhibitory factors are present in blood in vivo. RESULTS: No reduction in cellular cytotoxicity was found following treatment of effector cells with asialo GM1 antibody, but there was a reduction following treatment with asialo-GM1 antibody plus guinea pig serum and with guinea pig serum alone. Sera from pregnant, male, and SCID mice also reduced the level of cytotoxicity by metrial gland cell suspensions. CONCLUSIONS: The reduction in cytotoxicity in the presence of serum indicates that there are factors in the blood which are inhibitory to granulated metrial gland cell cytotoxic activity in vivo. The resistance of mouse metrial gland cells to asialo-GM1 antibody depletion is supportive of a natural cytotoxic (NC) lineage for this cell type. PMID- 10374705 TI - Oral vaccination of white-tailed deer using a recombinant Bacillus Calmette Guerin vaccine expressing the Borrelia burgdorferi outer surface protein A: prospects for immunocontraception. AB - PROBLEM: Reduction of excess numbers of white-tailed deer (Odocoileus virginianus) is a prime example of a potential use for immunocontraception as a means of wildlife population management. Oral vaccination appears to be the most pragmatic way to deliver immunocontraceptive vaccines to free-roaming populations of deer, but there was little, if any, prior evidence that oral vaccination is a viable concept in deer. METHOD OF STUDY: We used live Bacillus Calmette-Guerin (BCG) in a recombinant form (rBCG), which expressed Borrelia burgdorferi outer surface protein A, to test whether deer vaccinated orally with a specific antigen expressed in a live vector produce detectable antibody titers. RESULTS: The data indicate that oral vaccination of deer with an expressed antigen is feasible, as demonstrated by peak antibody titers to the expressed antigen. Also, peak titers measured by enzyme-linked immunosorbent assay were highest in orally vaccinated deer: 1600 in deer vaccinated by injection and 6400 in those vaccinated orally. CONCLUSIONS: The results of this study demonstrate that it is feasible to vaccinate deer orally with a live vector. PMID- 10374707 TI - Quantitative assessment of human leukocyte antigen-G protein in amniotic fluid by a double-determinant enzyme-linked immunosorbent assay using anti-human leukocyte antigen-G-specific antibody '87G'. AB - PROBLEM: The objective of this study was to establish an enzyme-linked immunosorbent assay (ELISA) system, in an attempt to quantify the amount of human leukocyte antigen (HLA)-G protein in amniotic fluid. METHOD OF STUDY: We established a double-determinant ELISA system using the anti-HLA-G specific mouse monoclonal antibody '87G' as a capture antibody and the horseradish-peroxidase labeled rabbit anti-human beta2-microglobulin antibody as a detection antibody. We then measured the concentration of HLA-G protein in amniotic fluid samples from nine normal second-trimester pregnant women and in serum samples from eight normal males. RESULTS AND CONCLUSION: HLA-G protein was detected in amniotic fluid at a concentration of 275 ng/ml (197-343 ng/ml) (median value and 95% confident range), whereas the concentration of HLA-G protein in male serum was below the minimum detection level. PMID- 10374708 TI - Yohimbine interacts with the dopaminergic system to reverse sexual satiation: further evidence for a role of sexual motivation in sexual exhaustion. AB - The possible interaction of yohimbine with the dopaminergic system in the mediation of sexual behaviour expression in sexually exhausted male rats was investigated. The behavioural effects of the simultaneous injection of yohimbine (500 microg/kg) plus apomorphine (50 microg/kg) and those of the combined treatment of haloperidol (125 microg), a nonspecific dopamine receptor antagonist, with an effective dose of yohimbine (2000 microg/kg) on sexually satiated rats were evaluated. Data show that yohimbine and apomorphine, per se, dose-dependently reverse sexual exhaustion by increasing the percentage of sexually satiated rats copulating and resuming copulation after ejaculation. Injection of haloperidol simultaneous to an effective dose of yohimbine, blocked the ability of the latter to reverse sexual satiation. The combined treatment with subthreshold doses of apomorphine and yohimbine synergised to reverse the sexual inhibition characteristic of sexual exhaustion. Data suggest that the dopaminergic system might be the final pathway for the yohimbine-induced sexual behaviour expression in satiated rats. The possible role of sexual motivation in the sexual exhaustion phenomenon is discussed. PMID- 10374709 TI - Involvement of alpha1-adrenoceptors in the basolateral amygdala in modulation of memory storage. AB - These experiments examined the involvement of alpha1-adrenoceptors in the basolateral amygdala and their interaction with beta-adrenoceptors in modulating memory storage. In Experiment 1, male Sprague-Dawley rats, implanted with bilateral cannulae in the basolateral amygdala, were trained in a one-trial inhibitory avoidance task and immediately after training, were given microinfusions (0.2 microl/side) of the selective alpha1-adrenoceptor antagonist, prazosin (0.1-1.0 microg). Retention was tested 48 h later. Prazosin induced a dose-dependent impairment in retention performance. In Experiment 2, animals received post-training intra-basolateral amygdala infusions of phenylephrine (a non-selective alpha-adrenoceptor agonist; 1.0-10.0 microg) alone or in combination with yohimbine (a selective alpha2-adrenoceptor antagonist; 0.2 microg) to examine the effects, on memory storage, of selective alpha1 adrenoceptor activation. Low doses of phenylephrine alone tended to impair retention performance, whereas the highest dose was non-effective. In contrast, phenylephrine infused together with yohimbine induced a dose-dependent enhancement of retention performance, suggesting that a selective activation of alpha1-adrenoceptors enhances memory formation. In Experiment 3, animals received intra-basolateral amygdala infusions of phenylephrine (1.0-10.0 microg) and yohimbine (0.2 microg) in combination with atenolol (a beta1-adrenoceptor antagonist; 1.0 microg). Atenolol blocked the memory-enhancing effects induced by infusions of phenylephrine together with yohimbine. Considered together, these findings suggest that alpha1-adrenoceptors in the basolateral amygdala are implicated in mediating the effects of norepinephrine on memory storage and that their action depends on concurrent beta-adrenoceptor activation. PMID- 10374710 TI - Venlafaxine: acute and chronic effects on 5-hydroxytryptamine levels in rat brain in vivo. AB - Venlafaxine is a dual serotonin (5-hydroxytryptamine, 5-HT) and noradrenaline uptake inhibitor which has been claimed to have an onset of antidepressant action which is faster than for other comparable drugs. The effects of venlafaxine on brain 5-HT levels in vivo have not yet been examined. Acute administration of venlafaxine to rats by i.p. injection resulted in dose-dependent increases in cortical and hippocampal 5-HT levels, as measured by in vivo microdialysis, over the range 5-20 mg/kg. The effect of venlafaxine (10 mg/kg i.p.) was potentiated by prior administration of pindolol (10 mg/kg s.c.) in hippocampus but not in frontal cortex. Daily administration of venlafaxine (5 mg/kg i.p.) for 4 weeks did not change basal 5-HT levels in either brain area. The effect of 8-hydroxy-2 (di-n-propylamino)tetralin (8-OH-DPAT, 0.2 mg/kg s.c.) to reduce 5-HT levels was unaffected by chronic venlafaxine at this dose, indicating that there was no change in sensitivity of presynaptic 5-HT1A autoreceptors. PMID- 10374711 TI - Differential involvement of mu-opioid receptor subtypes in endomorphin-1- and -2 induced antinociception. AB - We investigated the role of mu-opioid receptor subtypes in both endomorphin-1 and endomorphin-2 induced antinociception in mice using supraspinally mediated behavior. With tail pressure as a mechanical noxious stimulus, both intracerebroventricularly (i.c.v.) and intrathecally (i.t.) injected-endomorphins produced potent and significant antinociceptive activity. Antinociception induced by i.t. and i.c.v. injection of endomorphin-1 was not reversed by pretreatment with a selective mu1-opioid receptor antagonist, naloxonazine (35 mg/kg, s.c.). By contrast, antinociception induced by i.t. and i.c.v. endomorphin-2 was significantly decreased by mu1-opioid receptor antagonist. Antinociception of both i.t. and i.c.v. endomorphin-1 and -2 was completely reversed by pretreatment with beta-funaltrexamine (40 mg/kg, s.c.). The results indicate that endomorphins may produce antinociception through the distinct mu1 and mu2 subtypes of mu opioid receptor. PMID- 10374712 TI - Neuroprotection by the alpha2-adrenoceptor agonist, dexmedetomidine, in rat focal cerebral ischemia. AB - The present study was undertaken to explore the possible neuroprotective effect of the selective alpha2-adrenoceptor agonist, dexmedetomidine in a rat model of focal cerebral ischemia. The effect of dexmedetomidine (9 microg kg(-1)) on infarct volume was assessed and compared to that of glutamate receptor antagonists cis-4(phosphonomethyl)-2-piperidine carboxylic acid (CGS-19755) (20 mg kg(-1)) or 2,3-dihydro-6-nitro-7-sulfamoyl-benzo(F)quinoxaline (NBQX) (50 mg kg(-1)). Dexmedetomidine decreased total ischemic volume by 40% in the cortex (P<0.05) compared to NaCl-treated control rats, whereas NBQX reduced the infarct by 73% in the cortex (P<0.001) and by 43% in the striatum (P<0.01). Dexmedetomidine infusion was associated with some minor degree of hyperglycemia and hypotension. Drug-induced kidney changes were only seen in NBQX-treated rats. These results suggest that dexmedetomidine reduced ischemic volume despite causing a minor increase in blood glucose concentrations and hypotension. Its neuroprotective efficacy was better than that produced by CGS-19775, and dexmedetomidine was safer with respect to kidney toxicity when compared to NBQX. PMID- 10374713 TI - Relationship between myocardial cation content and injury in reperfused rat hearts treated with cation channel blockers. AB - A role for K+ and Ca2+ channel blockers in cardiac contractile dysfunction and myocardial ionic imbalance was examined in isolated rat hearts with 35-min ischemia and 60-min reperfusion. The K+ channel blockers glibenclamide (1-30 microM) and sematilide (1-30 microM), Ca2+ channel blockers diltiazem (0.1-3 microM) and nicardipine (0.03-1 microM) and fast Na+ channel blocker tetrodotoxin (0.01-0.3 microM) were delivered for the last 3-min pre-ischemia. Ischemia induced increase in Na+ content was attenuated by diltiazem and tetrodotoxin at all concentrations employed and by nicardipine at 0.3 microM, whereas the ischemia-induced loss of K+ was suppressed partially by glibenclamide and sematilide and almost completely by the two drugs in combination. Left ventricular developed pressure of untreated hearts did not recover upon reperfusion, which was associated with increases in myocardial Na+ and Ca2+ contents and decreases in K+ and Mg2+ contents. Glibenclamide and sematilide neither enhanced the post-ischemic recovery of left ventricular developed pressure nor affected cation changes during reperfusion. Diltiazem enhanced the recovery of left ventricular developed pressure and attenuated imbalance of the myocardial Na+ during ischemia and of all myocardial cations examined during reperfusion. The effects of nicardipine on these parameters were small. Tetrodotoxin enhanced the recovery of left ventricular developed pressure and reversed the imbalance of all myocardial cations examined during reperfusion in a concentration-dependent manner. The results suggest that blockade of transmembrane flux of K+ during ischemia plays a minor role in the improvement of post-ischemic contractile recovery, rather blockade of transmembrane flux of Na+ attenuates the ischemia and reperfusion injury. PMID- 10374714 TI - Characterisation of 5-HT receptors in human coronary arteries by molecular and pharmacological techniques. AB - 5-Hydroxytryptamine (5-HT) can produce both vasoconstrictor and vasorelaxant effects in human coronary arteries and the response to 5-HT can be influenced by the presence of disease. The aim of the present study was to elucidate the 5-HT receptor subtypes responsible for mediating 5-HT-evoked contraction of human coronary arteries using pharmacological, molecular and immunocytochemical approaches. Normal human coronary arteries, with intact endothelium, were mounted in tissue baths, and the vascular responses to 5-HT and 5-HT receptor agonists were studied. The effects of 5-HT1 and 5-HT2 receptor antagonists on these responses were also studied. Expression of messenger ribonucleic acid (mRNA) encoding different 5-HT receptors in human coronary arteries, atrium, ventricle wall and epicardium was determined using reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern blot analysis. The expression of 5-HT1B or 5-HT1D receptor protein was studied using subtype selective antibodies and standard immunocytochemical techniques. The rank order of 5-HT receptor agonist potency in causing vasoconstriction was 5-carboxamido tryptamine, (5-CT) > zolmitriptan = BW183C91 (N10-desmethyl zolmitriptan) = alpha-methyl-5-hydroxytryptamine (alpha CH3-5-HT) = 5-HT = sumatriptan > 2-methyl-5-hydroxytryptamine (2-CH3-5-HT) = 8 hydroxy-DPAT (8-OH-DPAT). Alpha-CH3-5-HT, 5-CT, 5-HT, zolmitriptan and BW 183C91 were significantly more potent (approximately 3-fold) than sumatriptan and 2-CH3 5-HT, which in turn were more potent than 8-OH-DPAT. Ketanserin and methiothepin (5-HT2 and 5-HT1 receptor antagonists, respectively) caused parallel rightward shifts of the concentration-effect curves to alpha-CH3-5-HT or 5-CT, respectively, without changing the maximum contractile response. In human coronary arteries, atrium. ventricle and epicardium. RT-PCR products corresponding to the human 5-HT2A, 5-HT1B and 5-HT1F receptors were expressed in high levels, mRNAs coding for 5-HT7, 5-HT1A and 5-HT1D receptors were only weakly expressed. No 5-HT1F receptor mRNA was detected. In coronary arteries there was a differential expression of 5-HT1B versus 5-HT1D receptor mRNAs, with 5-HT1B mRNAs being found in greater abundance. Dense 5-HT1B-immunoreactivity was detected on smooth muscle layer within coronary artery, however, 5-HT1D-immunoreactivity was not detected. It is concluded that 5-HT-evoked contraction of human coronary arteries is most probably mediated via the activation of both 5-HT1B and 5-HT2A receptors. PMID- 10374715 TI - Free radical involvement in endothelium-dependent responses of the rat thoracic aorta in moderate hypoxic conditions. AB - This study investigates the effects of agents which act on the production or efficacy of free radicals on the hypoxic responses of rat aorta rings. Under moderate hypoxic conditions, the resting tension of the rings was not changed but in rings precontracted with 5-hydroxytryptamine, there was a relaxation followed by a contraction. Removal of the endothelium with saponin suppressed relaxation to acetylcholine and abolished the contractions produced by hypoxia. In rings with a functional endothelium, hypoxic vasoconstriction was strongly inhibited by mannitol and exifone, but was not reduced by N(G)-nitro-L-arginine methyl ester, superoxide dismutase + catalase, or deferoxamine. Hypoxic vasodilatation was only partially inhibited by mannitol. To conclude, hypoxic constriction of the rat thoracic aorta is largely endothelium-dependent and involves free radicals whereas hypoxic dilatation is partially endothelium-dependent and partially involves free radicals. There is also indirect evidence for lack of direct involvement of nitric oxide/endothelium-derived relaxing factor (NO*/EDRF), hydroxyl radical (OH*) and superoxide anion in the hypoxic constriction and relaxation of the rat aorta. PMID- 10374716 TI - Role of protein kinase C and cAMP in fluoxetine effects on human T-cell proliferation. AB - In this work, we studied the effect of fluoxetine on human T-lymphocyte proliferation using optimal and suboptimal concanavalin A concentrations. In particular, we analyzed the influence of fluoxetine on the kinases that are involved in intracellular signalling after stimulation with mitogens. We found that fluoxetine promoted the Ca2+ -mediated proteolysis of protein kinase C (PKC) and increased cyclic-AMP (cAMP) levels, thereby impairing lymphocyte proliferation, when optimal concanavalin A concentrations were used. In contrast, when suboptimal concanavalin A concentrations were used, fluoxetine only increased PKC translocation, without modifying cAMP levels, leading to T-cell proliferation. According to our results, fluoxetine has a dual effect on T-cell proliferation by modulating the PKC and protein kinase A pathways. This mechanism is thought to be mediated through Ca2+ mobilization. PMID- 10374717 TI - Dexamethasone regulation of interleukin-1-receptors in the hippocampus of Theiler's virus-infected mice: effects on virus-mediated demyelination. AB - Intracerebral (i.c.) inoculation of susceptible strains of mice with Theiler's murine encephalomyelitis virus (TMEV) results in immune-mediated demyelinating disease. Interleukin-1 receptors are expressed in the brain of mice, in particular in the hippocampus, and have been implicated in neuroimmunoendocrine interactions. In the present study we investigated the regulation of interleukin 1 receptors in the hippocampus of a susceptible (SJL/J) and a resistant (BALB/c) strain of mice infected with TMEV, at different time intervals of the disease. Our results show that interleukin-1 receptors in the hippocampus were decreased in TMEV-infected mice at early times post-infection (10 and 14 days p.i.). The reduction in interleukin-1 receptors only occurred in the susceptible strain of mice (SJL/J), whereas interleukin-1 binding in the hippocampus of TMEV-infected resistant mice (BALB/c) showed values similar to those in control animals. The TMEV-induced down-regulation of interleukin-1 receptors was secondary to a marked decrease in the affinity of the receptor (control: Kd = 10.5 pM; TMEV: Kd = 1.30 pM) accompanied by a decrease in receptor number (control: Bmax = 2.189 fmol/mg protein; TMEV: B max = 0.84 fmol/mg protein). We also investigated the effects of glucocorticoid treatment on the regulation of hippocampal interleukin-1 receptors of TMEV-infected mice. Dexamethasone treatment in the early phase (500 microg/kg or 1 mg/kg during days 5-10 p.i.) of the disease significantly reversed the deficits in hippocampal interleukin-1 receptors observed at 10 days p.i. in SJL/J mice, and suppressed neurological signs of demyelination. These results suggest that: (i) the reduction of interleukin-1 receptors may be a consequence, at least in part, of local production of interleukin-1 at early times during TMEV infection; (ii) interleukin-1 seems to be a critical factor for the susceptibility to TMEV-induced demyelination and (iii) the protective effect of dexamethasone appears to be related to its ability to reverse the reduction in interleukin-1 receptors during the early disease. These results suggest that interleukin-1 is a pivotal mediator in TMEV-induced demyelination. PMID- 10374719 TI - Properties of amentoflavone, a potent caffeine-like Ca2+ releaser in skeletal muscle sarcoplasmic reticulum. AB - Amentoflavone, like caffeine, caused a concentration-dependent increase in Ca2+ release from the heavy fraction of fragmented sarcoplasmic reticulum of rabbit skeletal muscle. The Ca2+ -releasing activity of amentoflavone was approximately 20 times more potent than that of caffeine. The bell-shaped profile of Ca2+ dependence for amentoflavone was almost the same as that for caffeine. Typical blockers of Ca2+ -induced Ca2+ release channels, such as Mg2+, procaine and ruthenium red, inhibited markedly amentoflavone- and caffeine-induced 45Ca2+ release. The maximum 45Ca2+ release in response to amentoflavone was not changed by caffeine, but was further increased by adenosine-5'-(beta,gamma-methylene) triphosphate. This compound enhanced [3H]ryanodine binding to the heavy fraction of fragmented sarcoplasmic reticulum with a decrease in K(D) but without a change in Bmax. These results suggest that amentoflavone, which does not contain a nitrogen atom, probably binds to the caffeine-binding site in Ca2+ channels and thus potentiates Ca2+ -induced Ca2+ release from the heavy fraction of fragmented sarcoplasmic reticulum. PMID- 10374718 TI - Regulation of glomerular mesangial cell proliferation in culture by adrenomedullin. AB - Adrenomedullin is a recently discovered vasodilatory peptide that has been shown to be a potent activator of adenylate cyclase in a variety of cell systems, including rat mesangial cells. The major aim of the present study was to determine the regulation of rat mesangial cell proliferation (using [3H]thymidine incorporation as an index), apoptosis (using nucleosome-associated cytoplasmic DNA fragmentation as an index) and mitogen-activated protein kinase (MAPK) cascade, specifically extracellular signal-regulated kinase (ERK), jun-amino terminal kinase (JNK) and P38 mitogen-activated protein kinase (P38 MAPK) activities, by adrenomedullin-stimulated cyclic AMP-protein kinase-A pathway. Adrenomedullin increased cAMP levels significantly above basal and the response was inhibited by the adrenomedullin receptor antagonist, adrenomedullin-(22-52). Adrenomedullin also decreased [3H]thymidine incorporation and increased nucleosome-associated cytoplasmic DNA fragmentation, in a concentration-dependent fashion. Both these responses were receptor mediated as, adrenomedullin-(22-52) inhibited these effects. The decrease in proliferation and increase in apoptosis were both mimicked by forskolin, a direct adenylate cyclase activator. Adrenomedullin-mediated decrease in proliferation and increase in apoptosis were inhibited by H89 [[N-[2-((p-bromocinnamyl)amino)ethyl]-5-isoquinolinesulfonamide, hydrochloride]], a potent protein kinase-A inhibitor. Associated with the changes in proliferation and apoptosis, adrenomedullin decreased ERK2 activity, and increased JNK1 and P38 MAPK activities. All these kinase activities, except the increase in JNK1 activity could be simulated using forskolin. In addition, only adrenomedullin-mediated changes in ERK2 and P38 MAPK activities were inhibited by H89 while, adrenomedullin-stimulated JNK1 was not consistently inhibited by the protein kinase-A inhibitor. These results suggest that adrenomedullin might play an important role in mesangial cell turnover and that although adrenomedullin mediated responses are primarily cAMP-dependent, it does not preclude the involvement of cAMP-independent pathways. PMID- 10374720 TI - Effects of paxilline on K+ channels in rat mesenteric arterial cells. AB - The effects of paxilline, a mycotoxin, on whole-cell outward currents from freshly isolated cells of the rat mesenteric artery were studied. Paxilline inhibited a component of the outward current that was also sensitive to iberiotoxin. Inhibition could be observed at a concentration of 10 nM and complete inhibition of the iberiotoxin-sensitive current was achieved at 300 nM. The inhibition could be described by a single site of interaction with a Ki of 35.7 nM. Paxilline had no effect on the component of the current that was sensitive to 4-aminopyridine. It is concluded that paxilline is a potent inhibitor of large conductance Ca2+ -activated K+ currents in vascular smooth muscle cells. PMID- 10374721 TI - Functional alpha2C-adrenoceptors in human neuroblastoma SH-SY5Y cells. AB - The alpha2-adrenoceptor mediating inhibition of forskolin-stimulated cyclic AMP accumulation in human neuroblastoma SH-SY5Y cells was further characterized. The alpha2-adrenoceptor agonists, UK 14,304 (5-bromo-6-(2-imidazolin-2 ylamino)quinoxaline), oxymetazoline, guanfacine, (-)-noradrenaline and clonidine concentration-dependently decreased cyclic AMP accumulation in this cell line (Emax ca. 50% inhibition). Agonist pEC50 values ranged between 6.7 and 7.8. Clonidine was a partial agonist. The effects of UK 14,304 were blocked after a pertussis toxin treatment. The concentration-response curves of UK 14,304 were shifted to the right in a parallel manner by the following antagonists (mean pK(B) values): yohimbine (8.17), idazoxan (7.63), prazosin (6.66), 2-[2-(4-(2 methoxyphenyl)piperazin-1-yl)ethyl]-4,4-dimethyl-1,3-(2 H,4H) isoquinolindione (ARC 239; 7.12) and 2-(2,6-dimethoxyphenoxyethyl)aminomethyl-1,4-benzodioxane (WB 4101; 8.12). The relatively high pKB values of prazosin and ARC 239 point to a non-alpha2A-adrenoceptor-mediated effect. The relatively high pK(B) value of WB 4101 further characterizes the alpha2-adrenoceptor in SH-SY5Y cells as being of the alpha2C subtype. The analysis of the expression of alpha2-adrenoceptor subtypes by reverse transcriptase-polymerase chain reaction (RT-PCR) revealed the exclusive presence of alpha2C-adrenoceptor mRNA in SH-SY5Y cells. We propose that inhibition of forskolin-stimulated cAMP accumulation in SH-SY5Y cells be used as a functional model of human, native alpha2C-adrenoceptors. PMID- 10374722 TI - Optimization of cardiac fiber orientation for homogeneous fiber strain during ejection. AB - The strain of muscle fibers in the heart is likely to be distributed uniformly over the cardiac walls during the ejection period of the cardiac cycle. Mathematical models of left ventricular (LV) wall mechanics have shown that the distribution of fiber strain during ejection is sensitive to the orientation of muscle fibers in the wall. In the present study, we tested the hypothesis that fiber orientation in the LV wall is such that fiber strain during ejection is as homogeneous as possible. A finite-element model of LV wall mechanics was set up to compute the distribution of fiber strain at the beginning (BE) and end (EE) of the ejection period of the cardiac cycle, with respect to a middiastolic reference state. The distribution of fiber orientation over the LV wall, quantified by three parameters, was systematically varied to minimize regional differences in fiber shortening during ejection and in the average of fiber strain at BE and EE. A well-defined optimum in the distribution of fiber orientation was found which was not significantly different from anatomical measurements. After optimization, the average of fiber strain at BE and EE was 0.025 +/-0.011 (mean+/-standard deviation) and the difference in fiber strain during ejection was 0.214+/-0.018. The results indicate that the LV structure is designed for maximum homogeneity of fiber strain during ejection. PMID- 10374723 TI - Mechanics of leukocyte deformation and adhesion to endothelium in shear flow. AB - The mechanics of leukocyte [white blood cell (WBC)] deformation and adhesion to endothelial cells (EC) in shear flow has been investigated. Experimental data on transient WBC-EC adhesion were obtained from in vivo measurements. Microscopic images of WBC-EC contact during incipient WBC rolling revealed that for a given wall shear stress, the contact area increases with time as new bonds are formed at the leading edge, and then decreases with time as the trailing edge of the WBC membrane peels away from the EC. A two-dimensional model (2D) was developed consisting of an elastic ring adhered to a surface under fluid stresses. This ring represents an actin-rich WBC cortical layer and contains an incompressible fluid as the cell interior. All molecular bonds are modeled as elastic springs distributed in the WBC-EC contact region. Variations of the proportionality between wall shear stress (tau(w)) in the vicinity of the WBC and the resulting drag force (F(s)), i.e., F(s)/tau(w), reveal its decrease with WBC deformation and increasing vessel channel height (2D). The computations also find that the peeling zone between adherent WBC and EC may account for less than 5% of the total contact interface. Computational studies describe the WBC-EC adhesion and the extent of WBC deformation during the adhesive process. PMID- 10374724 TI - Determination of red blood cell velocity by video shuttering and image analysis. AB - A novel modification of conventional video imaging techniques has been developed to determine the velocity of red blood cells (RBCs), which offers compatibility with existing video-based methods for determining blood oxygenation and hemoglobin concentration. Traditional frame-by-frame analysis of video recordings limits the maximum velocity that can be measured for individual cells in vivo to about 2 mm/s. We have extended this range to about 20 mm/s, by electronic shuttering of an intensified charge-coupled device camera to produce multiple images of a single RBC in the same video frame. RBCs were labeled with fluorescein isothiocyanate and the labeled cells (FRBCs) were used as probes to determine RBC velocities in microvessels of the hamster retractor muscle. Velocity was computed as the product of the distance between centroids of two consecutive image positions of a FRBC and the shuttering frequency of the camera intensifier. In vitro calibrations of the system using FRBC and Sephadex beads coated onto a rotating disk yielded an average coefficient of variation of about 6%. Flow conservation studies at bifurcations indicated that the maximum diameter of microvessels below which all the FRBCs in the lumen could be detected was 50 microm. The technique was used to estimate mean-FRBC velocity distributions in vessels with diameters ranging from 8 to 50 microm. The mean-FRBC velocity profiles were found to be blunter than would be expected for Poiseuille flow. Single FRBCs tracked along an unbranched arteriole exhibited significant temporal variations in velocity. PMID- 10374725 TI - Theoretical gas phase mass transfer coefficients for endogenous gases in the lungs. AB - Gas phase mass transfer coefficients for nitric oxide (NO), ethanol (EtOH), and water vapor (H2O) were determined for typical conducting airway geometry and tracheal flows (5 x 10(-5)and 5 x 10(-4) m3 s(-1)), by solving the steady-state two-dimensional diffusion equation. A constant absolute production rate with first order consumption reactions in pulmonary tissue was assumed for NO. For EtOH and H2O, constant concentrations were assumed in the blood and tissue, respectively. Results, expressed in terms of the average Sherwood number (Sh), were correlated with the Peclet (Pe(r)) number, and the length-to-diameter (L/D) ratio for each airway branch in terms of a lumped variable, Pe(r)(L/D)n. (Sh) increases as the solubility of the gas in tissue and blood increases. In addition, Sh passes through a minimum value at Pe(r)(D/L)n equal to approximately one when axial convection and diffusion have equal but opposite magnitudes. We conclude that Sh is not a monotonic function of Pe(r)(L/D)n within the entire airway tree and that it depends on the physical properties of the gas in the tissue. This conclusion contrasts with previous experimental and theoretical correlations. PMID- 10374726 TI - Technique to determine inspiratory impedance during mechanical ventilation: implications for flow limited patients. AB - We present the design of an enhanced ventilator waveform (EVW) for routine measurement of inspiratory resistance (R) and elastance (E) spectra in ventilator dependent and/or severely obstructed flow-limited patients. The EVW delivers an inspiratory tidal volume of fresh gas with a flow pattern consisting of multiple sinusoids from 0.156 to 8.1 Hz and permits a patient-driven exhalation to the atmosphere or positive end-expiratory pressure. Weighted least-squares estimates of the coefficients in a sinusoidal series approximation of the EVW inspirations yielded inspiratory R and E spectra. We first validated the EVW approach using simulated pressure and flow data under different physiological conditions, noise levels, and harmonic distortions. We then applied the EVW in four intubated patients during anesthesia and paralysis: two with mild airway obstruction and two with severe emphysema and flow limitation. While the level of inspiratory R was similar in both groups of patients, the inspiratory E of the emphysematous patients demonstrated a pronounced frequency-dependent increase consistent with severe peripheral airway obstruction. We conclude that the EVW offers a potentially practical and efficient approach to monitor lung function in ventilator-dependent patients, especially those with expiratory flow limitation. PMID- 10374728 TI - Gene expression is altered in perfused arterial segments exposed to cyclic flexure ex vivo. AB - Certain regions of coronary and other arteries undergo cyclic flexure due to attachment to the heart or crossing of joints. Such motion gives rise to fluctuations in transmural stress and luminal shear stress. It is well known that cyclic variation of these biomechanical forces influences many aspects of vascular cell biology including gene expression. The purpose of this work was to investigate the hypothesis that cyclic flexure of arterial segments influences their gene expression. Bilateral porcine femoral arteries were obtained fresh from the abattoir. One vessel was mounted in an ex vivo perfusion system and subjected to an intraluminal pressure of 60 mmHg and flow of 50 ml/min to serve as a control. The other vessel was mounted in a second perfusion system with similar hemodynamic conditions, but also subjected to controlled cyclic bending consistent with that found in coronary arteries in vivo. Reverse transcriptase polymerase chain reaction analysis demonstrated that E-selectin and matrix metalloproteinase-1 (MMP-1) were consistently and significantly downregulated in the specimens subjected to 4 h of cyclic bending as compared to the control (n = 8, p < 0.05). Our results show that cyclic flexure of arterial segments in vitro may influence their gene expression. Further investigation should follow this novel observation and focus on other known mediators to more carefully elucidate the consequence of cyclic flexure on arterial pathobiology. PMID- 10374727 TI - Numerically based design of an orifice plate flowmetering system for human respiratory flow monitoring. AB - During certain medical procedures, it is important to continuously measure the respiratory flow of a patient, as lack of proper ventilation can cause brain damage and ultimately death. The monitoring of the ventilatory condition of a patient is usually performed with the aid of flowmeters. However, water and other secretions present in the expired air can build up and ultimately block a traditional, restriction-based flowmeter; by using an orifice plate flowmeter, such blockages are minimized. This paper describes the design of an orifice plate flowmetering system including, especially, a description of the numerical and computational techniques adopted in order to simulate human respiratory and sinusoidal air flow across various possible designs for the orifice plate flowmeter device. Parallel computation and multigrid techniques were employed in order to reduce execution time. The simulated orifice plate was later built and tested under unsteady sinusoidal flows. Experimental tests show reasonable agreement with the numerical simulation, thereby reinforcing the general hypothesis that computational exploration of the design space is sufficiently accurate to allow designers of such systems to use this in preference to the more traditional, mechanical prototyping techniques. PMID- 10374729 TI - Dynamics of the intrauterine fluid-wall interface. AB - Intrauterine fluid movements, which are responsible for embryo transport to a successful implantation site at the fundus, may be induced by myometrial contractions. Myometrial contractions in nonpregnant uteri were studied from in vivo measurements of intrauterine pressures with fluid-filled catheters and by visual observations of high-speed replaying of ultrasound images of the uterus. Transvaginal ultrasound (TVUS) images of sagittal cross sections of the nonpregnant uterus were scanned with an intravaginal ultrasound probe. Images at consecutive times (2 s apart) were digitized and processed by employing modern techniques of image processing. The sets of images were compared to evaluate time variation of the fluid-wall interface with respect to amplitude, frequencies, and wavelength of myometrial contractions. Analysis of TVUS images from 11 volunteers during the proliferative phase revealed that myometrial contractions are fairly symmetric and are propagated from the cervix towards the fundus at a frequency of about 0.01-0.09 Hz. The wavelength, amplitude, and velocity of the fluid-wall interface during a typical contractile wave were found to be 10-30 mm, 0.05-0.2 mm, and 0.5-1.9 mm/s, respectively. Additional data acquisition from a large number of normal subjects is needed to build a data base to predict normal characteristics of myometrial contractions in a nonpregnant uterus, in order to better understand their role in the preimplantation process. PMID- 10374730 TI - Dynamic details of disparity convergence eye movements. AB - Classically, the primary tool for quantifying the dynamics of vergence and other eye movements has been the main sequence. The main sequence is a plot of peak velocity versus response amplitude and is particularly useful for comparing the dynamics of a large number of eye movements over a range of response amplitudes. However, the main sequence represents only the equivalent first-order behavior of a response and does not describe its dynamics in detail. Since the main sequence is based on only two points on the dynamic trajectory, it is sensitive to measurement artifacts and noise. A new methodology is presented which quantifies the equivalent second-order dynamics of eye movements using a larger region of the transient response. These new indexes were applied to vergence eye movements and were found to differentiate between subtle, but important differences in movement dynamics. PMID- 10374731 TI - Principal dynamic mode analysis of nonlinear transduction in a spider mechanoreceptor. AB - The nonlinear dynamics of the mechanoelectrical transduction in an arthropod mechanoreceptor (cuticular slit sense organ of the spider Cupiennius salei) were studied using Volterra kernel measurements and the recently proposed method of principal dynamic modes. Since mechanoreceptors must operate with sufficient gain sensitivity to rapidly varying displacement stimuli over a broad bandwidth and for a wide range of amplitudes, the experimental data were generated by applying pseudorandom broadband mechanical displacements of various mean levels to the cuticular slits. The recorded response data were intracellular current and potential. The purpose of this paper is to demonstrate the use of the principal dynamic mode (PDM) methodology in elucidating the nonlinear dynamics of a spider mechanoreceptor. The results obtained demonstrate that two PDMs suffice to provide a complete nonlinear dynamic model of this insect mechanoreceptor. The first PDM resembles the first-order kernel and has a low pass characteristic (position dependent), while the second PDM has a high-pass characteristic (velocity-dependent) and resides entirely in the second-order kernel (nonlinear adaptation). This study may serve as an example of the proper use of this new methodology for the analysis of nonlinear physiological systems. PMID- 10374732 TI - Three-dimensional human head finite-element model validation against two experimental impacts. AB - The impact response of a three-dimensional human head model has been determined by simulating two cadaver tests. The objective of this study was to validate a finite-element human head model under different impact conditions by considering intracranial compressibility. The current University Louis Pasteur model was subjected initially to a direct head impact, of short (6 ms) duration, and the simulation results were compared with published experimental cadaver tests. The model response closely matched the experimental data. A long duration pulse was chosen for the second impact and this necessitated careful consideration of the head-neck joint in order to replicate the experimental kinematics. The skull was defined as a rigid body and was subjected to six velocities. Output from the model did not accurately match the experimental results and this clearly indicates that it is important to validate a finite-element head model under various impact conditions to define the range of validity. Lack of agreement for the second impact is attributed to the nonlinearity in the dynamic behavior of intracranial stress, a problem that is not reported in the literature. PMID- 10374733 TI - Chromosome Workshops 1998: current state of psychiatric linkage. PMID- 10374734 TI - Chromosomes 1, 2, and 7 workshop. PMID- 10374735 TI - Chromosome 4 Workshop Summary: Sixth World Congress on Psychiatric Genetics, Bonn, Germany, October 6-10, 1998. AB - This report summarizes the findings presented at the Chromosome 4 Workshop of the Sixth World Congress on Psychiatric Genetics (October 1998, Bonn, Germany). Chromosome 4 linkage and association results for several psychiatric phenotypes including bipolar affective disorder, schizophrenia, alcoholism, and mental retardation are reviewed. In bipolar affective disorder, positive linkage results for markers on 4q35 were reported by three independent groups. In addition, findings in bipolar disorder were reported for markers spanning 4p14-16, and of particular interest are the results that coincide with the original Blackwood et al. [1996: Nat Genet 12:427-430] region at 4p16. For schizophrenia, modest positive results were reported for 4q31, as well as for marker D4S2917 at a region of 4q close to the centromere. Chromosome 4 continues to demonstrate interesting results in alcoholism, particularly in the region of the alcohol dehydrogenase gene cluster; however, it is not clear how to interpret the contrast in the susceptibility versus protective loci that are being reported in this region. PMID- 10374736 TI - Report of the Chromosome 5 Workshop of the Sixth World Congress on Psychiatric Genetics. AB - The Chromosome 5 Workshop heard new data on a schizophrenia susceptibility locus in the 5q23.3-q31.1 region. Sixty-two pedigrees from Finland gave a lod score of 1.36 at the CSF1R locus approximately 14 cM distal to IL9/D5S393, where positive results from three pedigree collections converged at the 1997 workshop. Though positive at CSF1R, the new data were only weakly positive at the IL9 (lod 0.46) and D5S393 (lod 0.07) loci themselves. The workshop also reviewed new evidence in the 5p14.1-p13.1 region, where a large pedigree of schizophrenia of Puerto Rican extraction has suggested a susceptibility locus with a maximum lod score at D5S111. Twenty-one new pedigrees multiplex for schizophrenia in African Americans gave positive lod scores at D5S111 and flanking loci. In bipolar illness five genetically related pedigrees from the Saguenay-Lac-St. Jean region of Quebec identified a region of interest at 5q31.3-q35.1. This region overlaps with the D5S423 locus and includes the D5S812 locus and the 5q34 region, all of which are consistent with linkage in at least one other study. PMID- 10374737 TI - Chromosome 6 workshop report. AB - The Sixth World Congress on Psychiatric Genetics (Bonn, Germany, October 1998) provided a forum to review the linkage findings for psychiatric disorders on chromosome 6. Interest continues to center on 6p 24-22 for schizophrenia. Another area of possible linkage to both schizophrenia and bipolar disorder exists at 6q 21-22. Neither of these areas maybe considered confirmed at this time. PMID- 10374738 TI - Chromosomes 8 and 10 workshop. AB - The chromosomes 8 and 10 workshop took place at the Sixth World Congress on Psychiatric Genetics from October 6th-10th, 1998 in Bonn, Germany. Aim of the workshop was to discuss and summarize reports on potential susceptibility genes for psychiatric disorders. Linkage-findings on chromosome 8 concentrate on 8p22 p21 and include mainly schizophrenic disorders. Two areas on chromosome 10 were reported to contain potential susceptibility genes for schizophrenic as well as for affective disorders. The strongest findings were reported for 10p14-p11, while other groups communicated also linkage data for the telomeric part of the long arm to the workshop. PMID- 10374739 TI - Chromosome Workshop: chromosomes 11, 14, and 15. AB - This report describes linkage data presented at the Workshop on Chromosomes 11, 14, and 15 at the Sixth World Congress of Psychiatric Genetics in Bonn, Germany, together with relevant linkage data submitted to the chair and co-chair, and it is presented in the context of the previous literature concerning these chromosomes. We have attempted to collate current linkage data to provide a guide to potentially interesting findings on chromosomes 11, 14, and 15 for the phenotypes of bipolar disorder, schizophrenia, alcoholism, autism, and spelling and reading disability. We discuss methodological limitations and provide chromosome ideograms and tables summarizing findings to date. The most promising region currently appears to be 15q13-q15 in the region of the alpha 7 nicotinic receptor for the phenotype of schizophrenia (and, perhaps, more generally for functional psychosis). Additionally, 15q11-q13 in the region of GABRB3 holds interest as a potential site of a susceptibility gene for autism. Two regions on chromosome 11, 11p15 in the region of tyrosine hydroxylase gene and 11q22-q23 in the region of DRD2, continue to retain some interest for functional psychosis. PMID- 10374740 TI - Chromosomes 12 and 16 workshop. AB - Recent linkage results independently derived from a large French Canadian pedigree and Danish kindreds coupled with supportive data from other studies provide compelling evidence for a bipolar disorder susceptibility locus on chromosome 12q23-q24. The idea is further strengthened by the finding that Darier's disease, which maps to this region, has been shown to cosegregate with affective disorder in a family. This linkage finding, however, was not supported in other independent genome scans. On chromosome 16, bipolar families from Denmark exhibited suggestive linkage with D16S510, on 16p13. Multipoint nonparametric analysis on the NIMH Genetics Initiative bipolar pedigrees yielded increased allele sharing that maximized approximately 18 cM proximal to the latter locus. In contrast, evidence of linkage was not detected in other panels of bipolar families that were presented. At 16p13, a maximum multipoint lod score of 4 for a latent class-derived phenotype that has aspects of alcohol dependence was found in a genome scan of 105 families from the Collaborative Study of the Genetics of Alcoholism, identifying a potential vulnerability locus. PMID- 10374741 TI - Chromosome 13 workshop report. AB - Evidence was presented that provided support for linkage in a relatively broad telomeric region of chromosome 13. A significant overlap for positive markers linked to both bipolar disorder and schizophrenia occurred in this area. PMID- 10374742 TI - Report of the chromosome 18 workshop. AB - At the first chromosome 18 workshop held at the 1997 World Congress on Psychiatric Genetics (WCPG) in Santa Fe, NM, several studies were presented that suggested the presence of a bipolar disorder (BP) as well as a schizophrenia (SZ) susceptibility locus on chromosome 18. Although the fact that several independent studies all pointed to a susceptibility locus (or loci) on chromosome 18, the observation that these studies identified nonoverlapping candidate regions was disappointing at least from the viewpoint of molecular genetics aiming at cloning the respective gene(s). Together, the data suggested four possible regions of considerable size that contained a susceptibility gene. At the 1998 WCPG chromosome 18 workshop in Bonn, Germany, less data were submitted and with the exception of a few studies, most data were nonsupportive or negative. Although some refinements were made to the previous candidate loci, overall the picture has not changed in that we are still confronted with the same four potential loci on chromosome 18 for BP and/or SZ, i.e., 18p11.2 and 18q12.1-q12.3 for BP and SZ, and 18q21-q22 and 18q23-qter for BP. So far, no other psychiatric phenotypes show evidence for a susceptibility locus on chromosome 18. PMID- 10374743 TI - Chromosomes 19 and 20 report. PMID- 10374744 TI - Chromosome 21 workshop. AB - The report of the 1997 workshop presented overall evidence providing strong support for a susceptibility locus for bipolar disorder at C21q22-23. The 1998 workshop considered the latest results from four groups, and additional studies also have been incorporated into this report. The workshop noted that there was possibly a degree of overlap between the regions implicated by the large samples of the multiplex National Institute of Mental Health pedigrees (affected sib pair analysis: p = 0.0006) and the US/Israeli pedigrees of the New York group (admixture lod = 3.35), in an area a few centimorgans proximal to PFKL. Participants concluded that the evidence implicating this region remains as strong as any, and were optimistic that further investigation would eventually lead to the identification of a susceptibility gene. PMID- 10374745 TI - Chromosome 22 workshop report. AB - The chromosome 22 workshop took place at the Sixth World Congress on Psychiatric Genetics from October 6th-10th, 1998 in Bonn, Germany. Aim of the workshop was to summarize the findings in psychiatric genetics on chromosome 22. Four reports concerning a susceptibility locus for schizophrenia and one report on bipolar disorder were given. A potential locus for nocturnal enuresis has been suggested to reside on chromosome 22. PMID- 10374746 TI - Sixth World Congress of Psychiatric Genetics X Chromosome Workshop. AB - At the X chromosome workshop of the Sixth World Congress on Psychiatric Genetics, new data regarding psychiatric phenotypes and the X chromosome were presented. In the last year a number of groups have published linkage results for the X chromosome in schizophrenia, which provide no significant evidence for linkage. Presentations by groups from Cardiff, Oxford, State University of New York (SUNY), and Finland provide weak nonsignificant evidence for linkage of markers on the Xp11.4-p11.3, Xq21, and Xq26 with schizophrenia. However, the presence of a male-specific transmission ratio distorter (DMS1) that maps to Xp11.4-21.2 [Naumova et al., 1998: Am. J. Hum. Genet. 62:1493-1499] makes the interpretation of linkage findings in brother-brother pairs difficult in this region. Regarding bipolar affective disorder, little new data were reported, but previous reports provide evidence for linkage to Xq25-q26. Summary tables of linkage results for schizophrenia and bipolar disorder can be obtained from http://www.camh.net/ research/x-chromosome/. No linkage or transmission disequilibrium of polymorphisms of MAOA and MAOB in attention deficit hyperactivity disorder was seen. Negative results for transmission disequilibrium of polymorphisms of HTR2C and MAOA with autism were provided from German and Austrian families. PMID- 10374747 TI - The presence, origin, and significance of A beta peptide in the cell bodies of neurons. AB - Interest in the A beta amyloid in Alzheimer disease (AD) has largely focused on the A beta in the neuropil, an extracellular site. Here much attention has been given to the possibility that A beta acts as a neurotoxin. However, increasing emphasis is now being given to the relationship between neurofibrillary tangles (NFT) and the degree of cognitive decline, as opposed to the relationship between decline and senile plaques, the sites of extracellular A beta deposition. This review focuses attention on the existence and significance of A beta in the cell body of the neuron. The review brings together diverse strands of literature indicating: (1) the tau-positive, paired helical filaments that are the main component of NFT are not themselves the source of the amyloid-like staining (Congo red birefringence) of PHF, and are not, in fact, an "amyloid"; (2) there is A beta within the cytoplasm of neurons affected by AD and in other conditions characterized by tau-positive neurofibrillary tangles; (3) peptides derived from portions of the A beta precursor can bind to PHF; (4) the affected neurons are the source of extracellular A beta in their vicinity and are also unable to maintain the synaptic structures that depend upon the integrity of the neuronal cell body; and (5) debates about whether the intracellular A beta is an amyloid depend upon beliefs about its tertiary structure and assumptions concerning the relationship between the size of self-aggregating A beta molecules, their tertiary structure, and their ability to generate Congo red birefringence without necessarily being detected as ultrastructural filaments 5-10 nm wide. Based upon this literature, it is suggested that the Congo red birefringence generated by NFT is caused by A beta, intimately bound to the NFT. Moreover, whether defined as an amyloid or not, the A beta in the neuronal cell body indicates an abnormal processing of the precursor molecule on the way to its ultimate transmembrane domain. Deranged neuronal functioning, which leads to this abnormal processing and/or the intracellular A beta itself, may be the cause of subsequent functional and morphologic abnormalities in the brain. PMID- 10374748 TI - Increased interleukin-1beta converting enzyme expression and activity in Alzheimer disease. AB - Increased expression of interleukin-1 in Alzheimer disease (AD) has been implicated as a driving force in neurodegenerative cascades that underlie the formation of neuritic plaques and neurofibrillary tangles, the spread of these neuropathological lesions across cerebral cortical regions, and the accompanying neuronal cell injury and loss. The beta isoform of interleukin-1 is generated from an inactive, 33-kDa precursor through the action of a specific interleukin 1beta converting enzyme (ICE), also known as caspase-1. We used mesial temporal tissue (hippocampus and parahippocampal cortex) obtained postmortem from Alzheimer and control patients for determinations of endogenous tissue ICE activity and for Western immunoblot analysis of tissue ICE concentrations. ICE activity in Alzheimer tissue was elevated 50-100% (p < 0.002, or better, at incubation times of 30 min to 10 h), and ICE protein level was elevated 180% (p = 0.01). Parahippocampal cortex of Alzheimer patients showed increased numbers of neurons with in situ evidence of DNA damage. Increased DNA degradation was also evident upon electrophoresis of isolated DNA. Overexpression and increased activity of ICE may contribute to neurodegeneration in AD through generation of biologically active interleukin-1, with consequent activation of interleukin-1 driven neurodegenerative cascades. PMID- 10374749 TI - Evolution of neurofilament subtype accumulation in axons following diffuse brain injury in the pig. AB - Although accumulation of neurofilament (NF) proteins in axons has been recognized as a prominent feature of brain trauma, the temporal course of the accumulation of specific NF subtypes has not been well established. In the present study, 17 miniature swine were subjected to nonimpact inertial brain injury. At 3 hours (h), 6 h, 24 h, 3 days, 7 days, and 10 days post-trauma, immunohistochemical analysis was performed to determine axonal accumulation of NF-light (NF-L), the rod and sidearm domains and sidearm phosphorylation states of NF-medium (NF-M), and heavy (NF-H). We found that NF-L accumulation was easily identified in damaged axons by 6 h post-trauma, but NF-M and H accumulation was not clearly visualized until 3 days following injury. In addition, the axonal accumulation of NF-M and H appeared to be primarily comprised of the sidearm domains. While the accumulating NF was found to be predominantly dephosphorylated, we also detected accumulation of phosphorylated NF. Finally, we found that developing axonal pathology may proceed either towards axotomy with discrete terminal bulb formation or towards the development of varicose swellings encompassing long portions of axons. These findings suggest that there is a differential temporal course in NF subtype disassembly, dephosphorylation, and accumulation in axons following initial brain trauma and that these processes occur in morphologically distinct phenotypes of maturing axonal pathology. PMID- 10374750 TI - Versican enhances locomotion of astrocytoma cells and reduces cell adhesion through its G1 domain. AB - Versican is a large extracellular proteoglycan and is expressed in a variety of tissues including the central nervous system. A malignant astrocytoma cell line U87 with high motility expressed a higher level of versican than another malignant astrocytoma cell line U343 with lower motility. We observed that the U87 cells were less adherent to tissue culture plates than the U343 cells. To investigate the role of versican in astrocytoma cell migration, we generated recombinant products of a mini-versican construct expressed in COS-7 cells. We found that the mini-versican products enhanced astrocytoma cell migration. Furthermore, enhanced migration was promoted by the G1 domain but not the G3 domain of versican. We introduced culture medium containing products of the mini versican, the G1, and the G3 constructs separately into the astrocytoma cell lines U87 and U343. The mini-versican and the G1 construct, but not the G3 construct, were shown to reduce astrocytoma cell adhesion. The present data suggest that versican exerts its effect on astrocytoma cell migration and adhesion through the G1 domain. PMID- 10374751 TI - Identification of subgroups of high-grade oligodendroglial tumors by comparative genomic hybridization. AB - In contrast to astrocytic tumors, approximately two thirds of anaplastic oligodendrogliomas are reported to be chemosensitive. Relatively little is known about the genetic aberrations in oligodendroglial tumors (OTs). In order to elucidate oligodendroglial oncogenesis and to find specific genetic aberrations that may have prognostic and therapeutic implications, we performed comparative genomic hybridization (CGH) to detect chromosomal copy number changes in 17 low grade OTs (LG-OTs) and 12 high-grade OTs (HG-OTs) lacking a prominent astrocytic component. Loss of chromosome 1p (79%) and 19q (76%) were most frequently detected by CGH, all LG-OTs and 50% of the HG-OTs contained -1p (including 1p36 32), -19q (including 19q13.3), or both, and the rest of the HG-OTs showed +7, 10, or both. Since losses of 1p36-32 and 19q13.3 were mutually exclusive with +7 or -10, the HG-OTs could be divided in -1p/-19q and +7/-10 tumors. While the -1p/ 19q tumors can be considered as pure anaplastic oligodendrogliomas, the +7/-10 tumors may rather be glioblastomas with prominent oligodendroglial differentiation. We conclude that CGH is a powerful tool to assist in the identification of 2 major subgroups of HG-OTs with prognostic and possibly therapeutic relevance. PMID- 10374752 TI - Vascular endothelial growth factor (VEGF) modulates vascular permeability and inflammation in rat brain. AB - Vascular endothelial growth factor (VEGF) is an angiogenic growth factor that also induces vascular permeability and macrophage migration. VEGF expression is weak in normal adult brain, but is strongly upregulated in glioma cells and reactive astrocytes, suggesting that chronic overexpression of VEGF in the brain contributes to blood-brain barrier (BBB) breakdown. We examined the effects of chronic VEGF overexposure on the integrity of the BBB using the following approaches: 1) continuous intracerebral infusion of VEGF via miniosmotic pump; and 2) intracerebral injection of an adenoviral vector encoding the VEGF165 gene (AdCMV.VEGF). After 6 days both treatments produced approximately 10-fold breakdown of the BBB (as measured by transport of 14C-aminoisobutyric acid (AIB) from blood into brain) compared with the respective controls (albumin infusion or AdCMV.beta gal virus). BBB disruption in AdCMV.VEGF-treated brains was accompanied by a severe inflammatory response not observed in brains receiving AdCMV.beta gal or VEGF protein infusion, indicating that neither VEGF nor viral particles alone were responsible for the inflammatory response. However, injection of AdCMV.beta gal followed by VEGF infusion to the same site also elicited inflammation. Chronic overexposure of normal brain to VEGF also increased intercellular adhesion molecule-1 (ICAM-1) and major histocompatibility complex (MHC) class I and II expression. Although VEGF itself is not inflammatory, VEGF may modulate immune responses in the central nervous system (CNS) by opening the BBB, altering the immunoprivileged status of the brain, and allowing contact between normally sequestered CNS antigens and blood-borne immune mediators. PMID- 10374754 TI - Loss of proteins regulating synaptic plasticity in normal aging of the human brain and in Alzheimer disease. AB - Recent studies suggest that the cognitive impairment associated with normal aging is due to neuronal dysfunction rather than to loss of neurons or synapses. To characterize this dysfunction, molecular indices of neuronal function were quantified in autopsy samples of cerebral cortex. During normal aging, the most dramatic decline was found in levels of synaptic proteins involved in structural plasticity (remodeling) of axons and dendrites. Alzheimer disease, the most common cause of dementia in the elderly, was associated with an additional 81% decrease in levels of drebrin, a protein regulating postsynaptic plasticity. Disturbed mechanisms of plasticity may contribute to cognitive dysfunction during aging and in Alzheimer disease. PMID- 10374753 TI - Prosaposin gene expression and the efficacy of a prosaposin-derived peptide in preventing structural and functional disorders of peripheral nerve in diabetic rats. AB - We have recently demonstrated that prosaposin is a neurotrophic and myelinotrophic factor with the active trophic sequence located at the N-terminal region of the saposin C domain. There are also reports that prosaposin mRNA is increased distal to a physical nerve injury and that exogenous prosaposin treatment induces subsequent neuronal sprouting, suggesting involvement in repair processes. In the present study, we show that prosaposin mRNA is significantly (p < 0.05) elevated in the peripheral nerve of streptozotocin-diabetic rats, a model of insulin-deficient diabetes in which nerve injury arises from the metabolic trauma of hyperglycemia and its consequences. A 14 amino acid peptide derived from the neurotrophic region of prosaposin prevented the development of deficits in both large and small fiber function caused by diabetes in rats. The dose dependent prevention of nerve conduction slowing by TX 14(A) was accompanied by preservation of axonal caliber and sodium-potassium ATPase activity, while prevention of thermal hypoalgesia was associated with attenuation of the decline in nerve substance P levels. It is concluded that nerve subject to the metabolic injury of uncontrolled diabetes responds by increasing prosaposin gene expression, and that prosaposin-derived neurotrophic peptides may provide a novel therapeutic approach to treatment of diabetic and other peripheral neuropathies. PMID- 10374755 TI - An apoptotic depletion of oligodendrocytes in the twitcher, a murine model of globoid cell leukodystrophy. AB - Morphological alterations of oligodendrocytes (OLs) leading to their depletion were studied in the genetic demyelinating mutant, twitcher, a murine model of globoid cell leukodystrophy (GLD). With pi-glutathione-S-transferase immunostaining, OLs with multiple varicose processes were recognized in the early stages and adjacent areas of demyelination and then the OLs cytoplasm as well as the processes became shrunken with progression of the disease. These shrunken OLs were labeled by the TUNEL method, indicative of apoptotic cell death. The ultrastructural features of apoptotic cells were noted in these OLs and DNA laddering was noted in the twitcher brain in advanced stages. This is the first report describing the gradual depletion of OLs by apoptosis in genetic demyelination. PMID- 10374756 TI - Cell type specific upregulation of vascular endothelial growth factor in an MCA occlusion model of cerebral infarct. AB - Vascular endothelial growth factor (VEGF) is an endothelial cell specific mitogen that has been implicated in hypoxia-mediated angiogenesis under physiological and pathological conditions. We used the middle cerebral artery occlusion model (MCAO) in the rat to investigate VEGF mRNA and protein localization, and VEGFR-1 mRNA and VEGFR-2 mRNA expression in cerebral ischemia. By nonradioactive in situ hybridization we observed upregulation of VEGF mRNA and VEGFR-1 mRNA, but not of VEGFR-2 mRNA in the hemisphere ipsilateral to MCA occlusion. VEGF mRNA was upregulated in the periphery of the ischemic area commencing 3 hours (h) after onset of MCAO, reached a peak after 24 h, and remained expressed at lower levels until 7 days (d) after MCAO. Double labelling experiments revealed that the majority of VEGF expressing cells in the penumbra and within the infarct were immunoreactive for Ox-42, Iba-1, and Ed1, but not for GFAP and neurofilament proteins, suggesting that microglial cells/macrophages are the major cell type expressing VEGE Since VEGF was also expressed in Ox-42 immunoreactive cells distant from the infarct (e.g. in the corpus callosum and hippocampus), activated microglial cells expressing VEGF may migrate towards the ischemic stimulus. VEGF protein was also detected on capillaries within the peri-ischemic area, suggesting that VEGF produced and secreted by microglial cells/macrophages binds to its receptors on nearby vascular endothelial cells and initiates an angiogenic response which counterbalances tissue hypoxia. Accordingly, apoptosis of neuroectodermal cells in the penumbra was highly depressed after the onset of angiogenesis. The spatial and temporal correlation between the induction of angiogenesis with VEGF and VEGFR-1 expression suggests that the ischemic upregulation of VEGF represents a physiological response of the brain to counterbalance hypoxia/ischemia in order to protect neuroectodermal tissue. PMID- 10374757 TI - Frontotemporal dementia and corticobasal degeneration in a family with a P301S mutation in tau. AB - The tau gene has been found to be the locus of dementia with rigidity linked to chromosome 17. Exonic and intronic mutations have been described in a number of families. Here we describe a P301S mutation in exon 10 of the tau gene in a new family. Two members of this family were affected. One individual presented with frontotemporal dementia, whereas his son has corticobasal degeneration, demonstrating that the same primary gene defect in tau can lead to 2 distinct clinical phenotypes. Both individuals developed rapidly progressive disease in the third decade. Neuropathologically, the father presented with an extensive filamentous pathology made of hyperphosphorylated tau protein. Biochemically, recombinant tau protein with the P301S mutation showed a greatly reduced ability to promote microtubule assembly. PMID- 10374758 TI - The challenge of acute coronary syndromes. PMID- 10374759 TI - Acute coronary syndromes: biology. PMID- 10374760 TI - Acute coronary syndromes: diagnosis. PMID- 10374761 TI - Acute coronary syndromes: interventions. PMID- 10374762 TI - Acute coronary syndromes: drug treatments. PMID- 10374763 TI - If I had an acute coronary syndrome... PMID- 10374764 TI - ASAP is a bad idea. Atypical small acinar proliferation. PMID- 10374765 TI - The role of p21ras in pancreatic neoplasia and chronic pancreatitis. AB - K-ras mutations have been detected in both ductal cell carcinoma and intraductal papillary mucinous tumor (IPMT) of pancreas. The frequency of this mutation in ductal cell carcinoma is high, whereas in IPMT, it is variable. It has been suggested that the relatively high frequency of this mutation in ductal cell carcinomas compared with IPMT may account for the differences in biological behavior between these tumor types. More recently, the significance of K-ras mutations in pancreatic tissue has been questioned with the demonstration of this mutation in nonneoplastic pancreata. The current study aims to estimate the relative frequency and evaluate the biological significance of K-ras gene mutations in these neoplasms by performing polymerase chain reaction (PCR) assays of microdissected areas of IPMT, ductal cell carcinomas, and resected chronic pancreatitis. The study also investigates whether alterations of p21ras occur in K-ras mutation-negative cases by using immunohistochemical staining for K-, N- and H-ras. K-ras codon 12 mutations were found almost as frequently in IPMT (71%) as in ductal cell carcinomas (78%). They were also associated with the earliest morphological lesion, flat mucinous change. This mutation also was detected in 42% of cases of chronic pancreatitis. Expression of p21ras was found to correlate closely with K-ras mutation status in IPMT and ductal cell carcinoma. Negative staining for pan-ras, H-ras, and N-ras in cases with wild-type K-ras genes suggests that alternative routes of ras gene alteration are not operative in IPMT or ductal carcinoma. The findings suggest that K-ras activation is frequently associated with both IPMT and ductal cell carcinoma. Its high prevalence in nonneoplastic pancreata suggests that it is also associated with self-limited morphological lesions of the pancreas that do not progress to malignancy. PMID- 10374766 TI - Hepatocytic differentiation in retiform Sertoli-Leydig cell tumors: distinguishing a heterologous element from Leydig cells. AB - Sertoli-Leydig cell tumors (SLCT) of the ovary are rare sex cord-stromal neoplasms. A minority of SLCT are characterized by a pattern resembling that of the rete ovarii and frequently have a range of homologous and heterologous tissues. Approximately 20 cases of SLCT have been reported to have elevation of serum alpha-fetoprotein (AFP) levels, or tissue immunoreactivity for AFP, a protein usually associated with germ cell neoplasms, especially yolk sac tumor. We identified hepatocytic differentiation in five cases of retiform SLCT (RSLCT), and confirmed immunohistochemically that these cells are hepatocytes rather than Leydig cells. Hepatocytes are positive for keratins (AE1/3 and Cam 5.2), AFP, and ferritin, negative for vimentin, and show weak to moderate staining for inhibin. Leydig cells are negative for keratins, positive for vimentin, and intensely positive for inhibin. Immunohistochemistry is needed to distinguish hepatocytic differentiation from Leydig cells with certainty. Including the cases in this report, hepatocytic differentiation has been associated with a retiform pattern in SLCT in 14 of 25 cases (56%). The association of these two patterns appears to be characteristic of a relatively primitive sex cord-stromal neoplasm. PMID- 10374767 TI - Congenital cystic adenomatoid malformation of the lung (CCAM): evaluation of the cellular components. AB - Congenital cystic adenomatoid malformation of the lung (CCAM) is a rare congenital lesion whose pathogenesis is not well defined. It is generally accepted that the various types of CCAMs originate at different levels of the tracheobronchial tree. To further define the pathogenesis of CCAM, we evaluated the cellular composition of different CCAM types by immunohistochemistry. Twenty two CCAMs (17 CCAM type 1, two type 2, one type 3, and two type 4) were collected. The cellular composition was determined using immunohistochemical stains for type I cell-associated antigen (T1 cell-Ag), surfactant proteins and surfactant protein precursors (SP-A, SP-B, proSP-B, and proSP-C), neuroendocrine cells (GRP), Clara cells (UP-1), and the adhesion molecule CD44v6, a glycoprotein thought to be involved in cell-matrix and cell-cell interactions. Eleven fetal lungs also were analyzed to compare cytodifferentiation of the epithelial-lined cysts of the different types of CCAM with the stages of normal lung development. Our results indicate that CCAM is caused by an arrest in lung development, and, on the basis of cytodifferentiation, two major subtypes can be distinguished. One subtype consisting of CCAM types 1, 2, and 3 that shows a bronchiolar type of epithelium and a second subtype, consisting of CCAM type 4, that has an acinar alveolar type of epithelium. Our findings also suggest that these two subtypes may arise at different stages of the branching of the bronchopulmonary tree, the first at the pseudoglandular stage and the second at the saccular stage. PMID- 10374768 TI - Genetic features of Mexican women predisposing to cancer of the uterine cervix. AB - Cervical carcinoma is the most common neoplasia in Mexican women. Previous studies report association of this neoplasia with the major histocompatibility complex (MHC) antigens in Caucasians. In the present study, we compared antigen frequencies of class I and class II MHC phenotypes in patients and ethnically matched healthy controls. Patients had significantly increased frequencies of HLA A2 (PC = .000003) and HLA-DR5 (PC = .01) as compared with healthy controls. Conversely, we found a significant decrease of HLA-DR6 (PC = .01), HLA-DR2 (PC = .0005) and HLA-DR1 (PC = .0009) as compared with healthy controls. These results confirm some previous studies on HLA-associations with cervical carcinoma and reinforce the theory of independent mechanisms of MHC class I and class II genes in the etiopathogenesis of this disease. PMID- 10374770 TI - Angiomatoid neuroendocrine carcinoma of the thymus: report of a distinctive morphological variant of neuroendocrine tumor of the thymus resembling a vascular neoplasm. AB - Three cases of primary thymic neuroendocrine tumors characterized by prominent angiomatoid features that resembled a vascular neoplasm are presented. The patients were all men between 52 and 59 years of age who presented with chest pain and shortness of breath attributable to a large anterior mediastinal mass. The lesions ranged in size from 6 cm to 15 cm in greatest diameter, and were grossly soft and well circumscribed, but not encapsulated. The cut surface was remarkable for multiple blood-filled cyst-like spaces admixed with focal solid, hemorrhagic areas. Histologically, the tumors contained multiple cystically dilated spaces filled with blood which imparted the lesion with a striking angiomatoid appearance. The walls of the cysts were lined by a monotonous proliferation of round to oval cells with distinct cell borders, round central nuclei, and abundant eosinophilic cytoplasm. Mitotic activity was present in all cases and varied from 3 to 8 mitoses per 10 high-power fields. Immunohistochemical studies performed in two cases showed positivity of the tumor cells for keratin, Leu 7, and synaptophysin, and focal chromogranin positivity in one. Follow-up information obtained in two patients showed that both had died of tumor 4 and 8 years after initial diagnosis. The present cases show an unusual morphological appearance of thymic neuroendocrine tumors that may be mistaken for a vascular neoplasm. Immunohistochemical stains may be of importance in such instances in arriving at the correct diagnosis. PMID- 10374769 TI - Prognostic significance of microsatellite instability in sporadic mucinous colorectal cancers. AB - We investigated the prognostic significance of microsatellite instability (MI) in 50 consecutive patients with sporadic mucinous colorectal cancer who had undergone only surgery. We evaluated MI and the pathological features with a possible prognostic value for each tumor, and the effect of the examined parameters on patients' outcome was statistically analyzed (univariate and multivariate analysis). All patients were followed-up for a minimum of 72 months or until death; in evaluating survival, only deaths of colorectal cancer were considered. DNA extracted from tumor sections and the corresponding normal tissue was analyzed by polymerase chain reaction at six microsatellite loci: D2S123, D3S1611, D3S49, D5S107, BAT26, BAT40. Alterations at two or more loci were detected in 36% of cases (MI+ tumors). MI+ and MI- cancers differed significantly in the pattern of growth, and most MI+ tumors showed an expanding type of growth (72.2%, P = .005). At univariate analysis, improved survival rate was significantly associated with MI, as well as with the following parameters: expanding cancer growth, Dukes stage, and absence of venous invasion. Nevertheless, at multivariate analysis, only the pattern of cancer growth and Dukes stage were independent prognostic factors, whereas the effect on survival of MI and venous invasion was found to be negligible. In our study, MI+ and MI- cancers differ only on the pattern of growth; therefore, our data suggest that the better survival rate in mucinous cancers with genomic instability is strictly related to their less aggressive type of growth. PMID- 10374771 TI - Immunohistochemical markers of cell cycle control applied to ovarian and primary peritoneal surface epithelial neoplasms: p21(WAF1/CIP1) predicts survival and good response to platinin-based chemotherapy. AB - Immunohistochemistry for p53, p21(WAF1/CIP1), and Ki-67 provides insight into the molecular events controlling the cell cycle. We tested the hypothesis that these cell cycle markers will aid in the clinical evaluation of ovarian and primary peritoneal surface epithelial neoplasms (SENs). Paraffin sections from a retrospective surgical series of 117 SENs were immunostained with anti-p53 (clone DO7, Novacastra Laboratories, UK), anti-p21(WAF1/CIP1) (clone EA10, Oncogene Science, Cambridge, MA), and anti-Ki-67 (clone MIB-1, Immunotech, Westbrook, ME). The Ki-67 proliferation index (Ki-67PI) and immunoreactivity were evaluated. One hundred seventeen SENs reacted as follows: p53 50%+ and p21(WAF1/CIP1) 65%+. Ki 67PI ranged from 4% to 88% (mean/median = 44/46%). p53 reactivity associated with transitional cell histology, decreased p21(WAF1/CIP1) staining, increased Ki 67PI, architectural/nuclear grade, and stage (P < .05, 1 x 10(-7), .01, .05/.0001, .001,). p21(WAF1/CIP1) staining was associated with endometrioid/clear cell histology, decreased Ki-67PI, architectural/nuclear grade, and stage (P < 05/.05, .05, .01/1 x 10(-8), 1 x 10(-5)). Ki-67PI associated with increased architectural/nuclear grade but not mucinous histology (P < 1 x 10(-5)/1 x 10( 6), .01). Sixty-seven patients had disease at last follow-up; 53 were dead of disease at 0 to 67 months (mean/median, 21/18), and 14 were alive with disease at 12 to 224 months (mean/median, 56/40). Fifty patients were disease free at 5 to 214 months (mean/median, 59/41). Predictors of survival include decreased Ki 67PI, stage, architectural/nuclear grade (P < 1 x 10(-6), 1 x 10(-10), 1 x 10( 10)/.005) and p21(WAF1/CIP1) IMS (multivariate P < 1 x 10(-6)). p21(WAF1/CIP1), a potent inhibitor of cyclin-dependent kinases necessary for cell cycle progression, functions as a key checkpoint in cell cycle control. Immunoreactivity for p21(WAF1/CIP1) provides prognostic information independent of other histological and clinical predictors, p53 IMS, and Ki-67PI in this series of 117 PTs with SENs. Our preliminary data suggest an interrelationship between p21(WAF1/CIP1) expression and an effective clinical response to platinin based chemotherapy, both associated with apoptosis. Further investigation seems warranted. PMID- 10374772 TI - High expression of CD23 in the proliferation centers of chronic lymphocytic leukemia in lymph nodes and spleen. AB - Chronic lymphocytic leukemia (CLL) is a B-cell neoplasm composed of a heterogeneous mixture of cells, including small lymphocytes, prolymphocytes, and large transformed cells; these last cells appear to represent the proliferating compartment. CLL cells express, in addition to B cell markers, the transmembrane receptor CD23. CD23 functions as the receptor for IgE and also appears to play a role in controlling the growth and proliferation of lymphocytes. Its level of expression among the different cells in CLL has not been examined. In this study, we show that CD23 expression is much higher in the large transformed CLL cells than in the small lymphoid population. This may provide an explanation for the observed correlation between a circulating CD23 cleavage product (soluble CD23) and prognosis in CLL. In addition, we have shown that proliferation in splenic CLL occurs preferentially in the white pulp zones, even in cases in which both the white and red pulp are extensively infiltrated. PMID- 10374773 TI - Ventricular myocardium in control and growth-retarded human fetuses: growth in different tissue compartments and variation with fetal weight, gestational age, and ventricle size. AB - The aim of this study was to assess the growth of different tissue compartments in ventricular myocardium of control and intrauterine growth-retarded (IUGR) human subjects. Stereological counting and sizing methods were applied to cross sectional samples of hearts collected post mortem at 16 to 35 (control) and 32 to 42 (IUGR) gestational weeks. Total tissue volumes and total numbers of myocyte, connective tissue, and endothelial cell nuclei were estimated. In control hearts, the volume of each tissue compartment increased linearly over the period of gestation, and this was due to proliferation, because in each case the tissue volume per nucleus remained constant. In IUGR subjects, fetal weight, ventricle volume, myocyte volume, connective tissue volume, and endothelial nuclear number were less than expected for gestational age up to at least 35 weeks. Between this age and 8 postnatal weeks, these variables are predicted to achieve equivalence with values found in control fetuses. Similar deficits were found when variables were related to fetal weight, and it is predicted that equivalence with control values would be reached at weights of between 2.3 and 3.6 kg. No such differences between groups were detected when variables were related to ventricle size. These findings indicate that growth deficits in cardiomyocytes are accompanied, and may be influenced, by developmental delays that involve the intramyocardial interstitium (capillary bed, endothelium, and surrounding connective tissues). The delays and deficits exhibit "catch-up" to control values between 35 weeks and term. PMID- 10374774 TI - Loss of heterozygosity of the Wilms' tumor suppressor gene (WT1) in in situ and invasive breast carcinoma. AB - The Wilms' tumor suppressor gene (WT1), a nuclear transcription factor, regulates the expression of the insulin-like growth factor (IGF) and transforming growth factor (TGF) systems, both of which are implicated in breast tumorigenesis. WT1 allelic integrity was examined by loss of heterozygosity (LOH) studies in formalin-fixed, paraffin-embedded (FFPE) ductal carcinoma in situ (DCIS, n = 20) and fresh frozen primary invasive breast carcinomas (n = 24). Loss of heterozygosity (LOH) at the WT1 locus (11p13) was examined by PCR evaluation of an Hinf1 restriction fragment length polymorphism (RFLP) and correlated to tumor stage (in situ and invasive). After identification of the heterozygous/informative breast lesions, 1 of 12 (8.3%) DCIS (high-grade micropapillary) and 3 of 14 (21.4%) of infiltrating carcinomas (high grade) showed loss of one allele, suggesting that LOH of the WT1 locus is a rare genetic event in early breast cancer, becoming more common in invasive and in high-grade lesions. PMID- 10374775 TI - Pulmonary pathology in Gaucher's disease. AB - Gaucher's disease is a familial storage disease caused by a deficiency of the enzyme glucocerebrosidase. Pulmonary involvement is considered rare in Gaucher's disease, especially type I. Sporadic case reports have shown various types of lung involvement, but the spectrum of pulmonary pathology in Gaucher's disease has not been described. Nine cases of Gaucher's disease were retrieved from the autopsy file of Hadassah Medical Center, Jerusalem, Israel. There were six cases with type I Gaucher's disease and three cases with type II. Lung sections were evaluated, and special stains were employed, including immunohistochemical stains for CD68, cytokeratin, and CD34. Gaucher cells were found in the lungs in all nine cases. The involvement was considered pathologically significant in five of nine cases and clinically significant in three of nine cases. Four distinct patterns of pulmonary involvement by Gaucher cells emerged: intracapillary (9 of 9), patchy interstitial infiltrates in a lymphatic distribution (2 of 9), massive interstitial thickening of alveolar septa (1 of 9), and intra-alveolar infiltrates (2 of 9). The universal involvement of pulmonary capillaries indicates that this is probably systemic in nature and not intrinsic to the lungs. Pulmonary involvement in Gaucher's disease is commoner than previously recognized. Immunocytochemical stains help to identify isolated Gaucher cells and distinguish them from native alveolar macrophages. PMID- 10374776 TI - Inhibin and CD99 (MIC2) expression in uterine stromal neoplasms with sex-cord like elements. AB - Uterine mesenchymal neoplasms with sex-cord-like elements are designated as endometrial stromal tumor with sex-cord-like elements (ESTSCLE) or uterine tumor resembling ovarian sex-cord tumor (UTROSCT), depending on the extent of sex-cord like differentiation. Occasionally, sex-cord elements similar to those in ESTSCLE and UTROSCT occur in uterine adenosarcomas. To determine whether the sex-cord like elements in these tumors show immunohistological evidence of sex-cord differentiation, we studied a series of uterine neoplasms for expression of inhibin, a peptide hormone expressed by normal ovarian granulosa cells and ovarian sex-cord neoplasms, and CD99, a protein also expressed by granulosa cells, Sertoli cells, and some ovarian sex-cord tumors. Thirty uterine mesenchymal neoplasms (five epithelioid or plexiform smooth muscle tumors, three endometrial stromal tumors, two mixed endometrial stromal and smooth muscle tumors, 10 ESTSCLE, five UTROSCT, and five miscellaneous stromal processes) and five epithelial neoplasms were evaluated for expression of CD99 (clone 12E7) and inhibin (clone R1) in formalin-fixed, paraffin-embedded tissue. Three of 10 (30%) ESTSCLE and five of five (100%) UTROSCT were inhibin and CD99 immunoreactive. Inhibin staining was confined to the areas with sex-cord-like differentiation, and staining was generally much stronger and more extensive in areas featuring prominent foam cells. There were no differences in the degree or intensity of staining for inhibin in premenopausal and postmenopausal women. CD99 expression tended to correlate with inhibin and was typically confined to similar cell types in the individual neoplasms. Weak CD99 immunoreactivity was seen in one additional epithelioid smooth muscle tumor, whereas all other mesenchymal and epithelial neoplasms studied for inhibin and CD99 were negative. These results provide further immunohistological support for true sex-cord differentiation within uterine mesenchymal proliferations and suggest that the degree of sex-cord differentiation may correlate with the expression of these markers. PMID- 10374777 TI - Expression of cathepsin B and cystatin C in human colorectal cancer. AB - Cathepsin B is a matrix protease that may be associated with colorectal carcinoma invasion and progression. In this study, we investigated the localization of cathepsin B in cancerous and noncancerous tissues of 80 patients with colorectal cancer including 25 cases with liver metastasis. In addition, the expression of cystatin C, one of several cathepsin B inhibitors, was compared with that of cathepsin B in the same samples to reveal one of the regulation mechanisms of cathepsin B. The cancer cells in the advancing edge of the tumors often exhibited the strongest immunostaining of cathepsin B, and stromal cells and normal epithelial cells adjacent to the tumors were also positive for cathepsin B. The percentage of cathepsin B-positive cases was significantly larger in the group with liver metastases than in the group without liver metastases. In the group without liver metastases, the cancer cells and stromal cells more frequently exhibited cathepsin B immunoreactivity in Dukes' A cases than in Dukes' B and C cases. In situ hybridization and reverse transcription-polymerase chain reaction (RT-PCR) confirmed cathepsin B synthesis in the cancer and proximal epithelial cells. There was an average 3.7-fold increase in cathepsin B mRNA levels in the cancerous tissues compared with that of noncancerous tissues, and Dukes' A tumors exhibited the highest expression level. Conversely, cystatin C mRNA levels were similar in all samples, and tended to show an inverse correlation with the cathepsin B levels. In conclusion, cathepsin B expression by human colorectal cancers and surrounding noncancerous cell components may contribute to both local invasion at the early stage and remote metastasis without influence of cystatin C. PMID- 10374778 TI - Placental site nodule and characterization of distinctive types of intermediate trophoblast. AB - Both placental site nodule and exaggerated placental site are described as being composed of intermediate trophoblast (IT), yet their morphological features and clinical presentation differ significantly. This study was undertaken to evaluate the morphological and immunohistochemical features of trophoblastic cells in placental site nodules and compare them with the trophoblastic cells in exaggerated placental sites as well as in different anatomic locations in the developing placenta to evaluate these differences. Forty-two placental site nodules, 20 abortus specimens ranging from 3 to 13 weeks, 8 second- and 10 third trimester placentas, and 12 exaggerated placental sites were studied by conventional light microscopy and immunohistochemistry. This analysis showed that the trophoblastic cells in the placental site nodule closely resemble those in the chorion laeve. We have designated these cells "chorionic-type IT cells." They are composed of two populations of cells, one with eosinophilic and the other with clear (glycogen-rich) cytoplasm. The eosinophilic cells tended to be larger with more pleomorphic nuclei, whereas the clear cells were smaller with more uniform nuclei. Chorionic-type IT cells in the chorion laeve and placental site nodule were diffusely positive for placental alkaline phosphatase but were only focally positive or negative for human placental lactogen (hPL), Mel-CAM (CD146), and oncofetal fibronectin. In contrast, hPL, Mel-CAM, and oncofetal fibronectin were diffusely expressed in IT cells in the placental site, both normal and exaggerated. The chorionic-type IT cells in placental site nodule and chorion laeve showed mild proliferative activity as indicated by an increased Ki-67 labeling index (3% to 10%). In contrast, the Ki-67 labeling index in normal and exaggerated implantation sites was zero. The morphological and immunohistochemical features of chorionic-type IT cells contrast with the IT cells in the implantation site that we have designated "implantation site IT cells." Both types of IT cells develop from a population of trophoblastic cells in the trophoblastic columns that we have tentatively termed "villous IT cells." Four of 42 placental site nodules were larger (>5 mm) than the remainder and showed transitional features between a typical placental site nodule and an epithelioid trophoblastic tumor, a recently described distinctive gestational trophoblastic tumor. There were no recurrences among the placental site nodules regardless of size. All placental site nodules were immunoreactive for inhibin alpha and cytokeratin 18, whereas 33 squamous cell carcinomas of the cervix, which can at times be confused with placental site nodules, were negative. In conclusion, there appear to be three subpopulations of IT cells with distinctive morphological and immunohistochemical features. Different subpopulations can be related to different trophoblastic lesions: implantation site IT cells to an exaggerated placental site and its neoplastic counterpart, placental site trophoblastic tumor and chorionic-type IT cells to a placental site nodule and its neoplastic counterpart, epithelioid trophoblastic tumor. PMID- 10374779 TI - Utility of surfactant protein B precursor and thyroid transcription factor 1 in differentiating adenocarcinoma of the lung from malignant mesothelioma. AB - Differentiation of malignant mesothelioma from adenocarcinoma, particularly from a lung primary, remains a difficult diagnostic problem. Surfactant protein B precursor (pro-SP-B) and thyroid transcription factor 1 (ITF-1) are expressed selectively in the normal respiratory epithelium and in adenocarcinomas of the lung. In this study, we evaluated the utility of pro-SP-B and ITF-1 in distinguishing pulmonary adenocarcinomas and malignant mesotheliomas. Immunoreactivity for pro-SP-B and TTF-1 was examined in paraffin sections of 370 primary lung carcinomas (208 adenocarcinomas, 101 squamous cell carcinomas, and 61 large cell carcinomas) and 95 malignant mesotheliomas, using a pro-SP-B antiserum and a monoclonal TTF-1 antibody with a biotin-streptavidin detection system. Immunostaining for pro-SP-B was detected in 57% of adenocarcinomas, and 20% of large cell carcinomas. Immunoreactivity for TTF-1 was shown in 76% of adenocarcinomas and 26% of large cell carcinomas. Malignant mesotheliomas and squamous cell carcinomas did not stain with either antibody. The expression of pro-SP-B and TTF-1 in adenocarcinomas of the lung but not in malignant mesotheliomas shows that pro-SP-B and TTF-1 staining is useful in differentiating these neoplasms. PMID- 10374780 TI - Bcl-2 and Bcl-X expression in gangliogliomas. AB - Bcl-2 and bcl-xL are proteins known to inhibit cell death (apoptosis). Expression of these proteins in gangliogliomas has not been extensively examined. This study retrospectively evaluates bcl-2 and bcl-x immunostaining in paraffin-embedded materials in gangliogliomas. Twenty-nine gangliogliomas in 17 males and 12 females, age 2.5 to 47 years (mean, 20.7 years), were studied. Nineteen tumors were situated primarily in the temporal lobe. All but three patients presented with seizures ranging from 3 months to 28 years' duration (mean, 11.1 years) before surgery. All tumors histologically were comprised of an atypical neuronal component and a glioma component, which most frequently resembled a low-grade astrocytoma. Cortical dysplasia was observed adjacent to eight tumors. MIB-1 (marker of cell proliferation) labeling indices (percentage of positively staining tumor cell nuclei) ranged from 0 to 7.7 (mean, 0.8). bcl-2 staining was observed in 25 tumors (86%); neuronal staining was present in 24 cases (83%), and glial cell staining in 21 tumors (72%). Bcl-xL staining was only observed in eight gangliogliomas (28%); in all eight tumors (28%), neuronal staining was seen, and focal glial cell staining was present in two cases (7%). Four tumors (14%) did not stain with either bcl-2 or bcl-xL. There appeared to be no relationship between MIB-1 immunostaining and staining with bcl-2 or bcl-xL. bcl 2 expression by immunohistochemistry was observed more frequently than bcl-xL in gangliogliomas. Expression of these proteins may reflect abnormalities of apoptosis, which could play a role in the survival of cells that may be involved in the development of gangliogliomas. PMID- 10374781 TI - Trisomy 3 in gastric lymphomas of extranodal marginal zone B-cell (mucosa associated lymphoid tissue) origin demonstrated by FISH in intact paraffin tissue sections. AB - Some reports have indicated that trisomy 3 represents a characteristic chromosomal abnormality found in lymphomas arising in mucosa-associated lymphoid tissues (MALT)/extranodal marginal zone B-cells (MZBC). Traditional cytogenetic analysis of metaphase preparations is cumbersome and not always possible, especially in those situations in which the diagnosis in not suspected before a biopsy. Our aim is to use a relatively simple method to evaluate trisomy 3 in paraffin-embedded, formalin-fixed tissue, using fluorescence in situ hybridization (FISH) on intact tissue sections. Formalin-fixed, paraffin-embedded archival tissues from 30 cases (27 lymphoma and 3 chronic gastritis cases) were hybridized with a chromosome 3 specific alpha-satellite probe (ONCOR, Gaithersburg, MD). Three of four cases of gastric MZBC/MALT lymphoma revealed trisomy 3. Ten cases of lymphoma of possible or probable MZBC origin were examined, and four revealed trisomy 3. Five of 13 non-MZBC lymphomas revealed trisomy 3. None of the chronic gastritis cases nor normal tonsil cases revealed trisomy 3. Our results, using a different methodological approach, confirm the findings of others that trisomy 3 is an abnormality found in a significant proportion of lymphomas of MZBC origin. Our approach also makes possible interphase cytogenetic analysis (by FISH) of routinely processed formalin-fixed, paraffin-embedded tissues, without the need to disaggregate cells. It thus may facilitate genetic analysis on specimens previously deemed unsuitable for such analysis, particularly when tissue quantity is limited. PMID- 10374782 TI - Survival in small cell lung carcinoma is independent of Bcl-2 expression. AB - Bcl-2 overexpression is a common event in small cell carcinomas (SCLC). The bcl-2 oncoprotein has a unique oncogenic role by inhibiting programmed cell death (apoptosis), resulting in tumorigenesis and chemoresistance. Forty-two cases of SCLC were stained immunohistochemically with bcl-2 monoclonal antibody (Biogenex, San Ramon, CA) after using an antigen retrieval step with citrate buffer. bcl-2 positivity was determined as detection of the oncoprotein in greater than 10% of noncrushed neoplastic cells. Twenty-five of 42 (60%) patients had extensive disease at presentation, 10 of 42 (24%) had limited disease, and 7 of 42 (16%) had disease localized to the lung. Twenty-four of 42 (57%) tumors were bcl-2 positive, and 18 of 42 (43%) tumors were bcl-2 negative. Follow-up in patients ranged from 7 days to 96 months (mean follow-up, 20 months). The median survival of patients with bcl-2-positive tumors was 11 months, as opposed to 13 months for bcl-2-negative tumors. There was no significant difference in median survival between bcl-2-positive and bcl-2-negative SCLC (log rank test, P = .2256). Using Cox's proportional hazards model, median survival in SCLC was determined to be independent of age at diagnosis, stage at presentation, therapeutic modality, and bcl-2 expression. bcl-2 expression does not significantly influence survival in SCLC. PMID- 10374783 TI - Granulocyte colony-stimulating factor-producing primary pericardial mesothelioma. AB - A 54-year-old male patient presented with a granulocyte colony-stimulating factor (G-CSF)-producing primary pericardial mesothelioma, while showing symptoms of congestive heart failure, a fever of 38 to 39 degrees C, and marked leucocytosis of 52.7 x 10(3) cells/mm3. The histopathologic diagnosis was established after autopsy. G-CSF production was confirmed by the expression of G-CSF mRNA in the tumor extract and the patient's high serum G-CSF concentration. The expression of G-CSF by benign and malignant mesothelial cells has already been reported. However, this is the first case report of G-CSF production in a pericardial mesothelioma. PMID- 10374784 TI - Fatal poisoning from liquid dimethylmercury: a neuropathologic study. AB - Since ancient times, mercury has been recognized as a toxic substance. Dimethylmercury, a volatile liquid organic mercury compound, is used by a small number of chemistry laboratories as a reference material in nuclear magnetic resonance spectroscopy. To our knowledge, dimethylmercury has been reported in only three cases of human poisoning, each proving fatal. Very small amounts of this highly toxic chemical can result in devastating neurological damage and death. We report the neuropathologic findings in a fatal case of dimethylmercury intoxication occurring in a laboratory researcher that resulted from a small accidental spill. We compare these findings to those reported in one previously reported fatal case of dimethylmercury poisoning, and to earlier reports of monomethylmercury poisoning, and discuss the clinicopathologic correlation. PMID- 10374785 TI - Chordoid glioma of the third ventricle: confirmatory report of a new entity. AB - The term "chordoid glioma" was recently introduced to denote a circumscribed, apparently low-grade neoplasm arising in or preferentially involving the third ventricle of middle-aged women. We report biopsy and postmortem findings in a 60 year-old woman with symptoms of forgetfulness, headache, and lethargy. Neuroimaging showed a contrast-enhancing third ventricular mass with obstructive hydrocephalus. The tumor was subtotally resected. Microscopically, it consisted of clusters and strands of epithelioid cells in a mucoid matrix. Its margins were remarkably discrete and showed little tendency to infiltrate surrounding brain parenchyma. The majority of neoplastic cells were glial fibrillary acidic protein (GFAP) and vimentin positive, whereas S100 protein labeled only individual cells. Stains for epithelial membrane antigen (EMA) and cytokeratin were nonreactive. There was no evidence of neuroendocrine differentiation or expression of estrogen and progesteron receptors. Lymphoplasmacellular infiltrates were noted throughout the lesion and at the tumor-brain interface. The MIB-1 labeling index averaged 1.5%. At present, chordoid glioma is considered a glial neoplasm of uncertain histogenesis with distinct clinicopathologic features. PMID- 10374786 TI - Mast cell disease and malignant germ cell tumors. PMID- 10374787 TI - Prostatitis unplugged? Prostatic massage revisited. AB - Repetitive prostatic massage is not a new tool in the urologists' armatarium. Once the most popular therapeutic maneuver used to treat prostatitis, it was abandoned as primary therapy almost 3 decades ago. Based on experience reported outside North America and anecdotal experiences of some patients and their physicians, it may be making a comeback. Unfortunately, there are almost no prospective data that would substantiate a claim as to its effectiveness. This article discusses the historic aspects of prostatic massage, suggests possible mechanisms of action, and describes the opinions of North American urologists who are associated with academic clinical research centers and are universally acknowledged as experts in the diagnosis and treatment of prostatitis. At this time, the science of prostatic massage must rely on anecdotal experiences, small, uncontrolled studies, and perhaps somewhat biased opinions of the major thought leaders in the field of chronic prostatitis/chronic pelvic pain syndrome. PMID- 10374788 TI - Vasovasostomy: Macroscopic approach and retrospective review. AB - The objective of this study was to review our 5-year experience with macroscopic vasovasostomy. A series of 160 consecutive patients who underwent macroscopic vasovasostomies between 1991 and 1996 at Madigan Army Medical Center were studied. A modified one-layer anastomotic technique using 7-0 or 8-0 Prolene sutures was utilized. Results showed that 87% of patients who underwent bilateral vasectomy reversal showed return of sperm to the ejaculate with a mean progressive sperm motility of 32.2%. Clinical success was measured by successful pregnancies in 50% of patients in whom <5 years had elapsed since vasectomy. The average operative time was 1 hour 15 minutes. Although slightly better results have been obtained using the microscopic approach, vasovasostomy performed under loupe magnification has the advantage of reduced cost, decreased operative time, and a lower operator skill level. PMID- 10374789 TI - Outcome analysis of minimally invasive treatments for benign prostatic hyperplasia. AB - The armamentarium of minimally invasive treatment modalities for patients with benign prostatic hyperplasia has increased steadily during the past decade. The energy sources used range from microwaves and radiofrequency waves to high intensity focused ultrasound, with laser vaporization/coagulation/resection and electrosurgical techniques. The large amount of data available allow some conclusions to be drawn concerning the present role of the "gold standard" TURP among the minimally invasive procedures. Although the subjective response after TURP and other minimally invasive procedures is comparable, improvements of flow and urodynamic parameters usually are more pronounced after TURP. Failure rates requiring reintervention (usually TURP) are considerable. Minimally invasive procedures lead to a shift of morbidity from the intraoperative phase, which is reduced (risk of bleeding, TUR syndrome, transfusion) to the postoperative phase. This period is characterized by prolonged urinary retention (ILC, VLAP), significant dysuria (VLAP, TUVP), and nycturia. Recent advances in electrosurgical techniques, such as band TURP loops that facilitate coagulation due to the longer contact time between the electrode and the tissue, have the potential to convert TURP into a less invasive procedure. Finally, high-energy TUMT seems to offer a truly minimally invasive treatment combining efficacy and the need for topical anesthesia only. However, due to a lack of homogeneity of criteria for patient recruitment, parameters of evaluation, and adequate follow up; accurate guidelines for appropriate patient management have not been established yet. PMID- 10374790 TI - Stamey needle passage of fascial pubovaginal sling after failed vesicourethropexy. AB - In cases of recurrent stress urinary incontinence after failed anti-incontinence surgery, it is common for the bladder neck and anterior urethra to be fixed to the symphysis pubis, increasing the risk of inadvertent bladder perforation during reoperation. We describe a modification to the popular pubovaginal sling using a 15 degrees angled Stamey suspension needle for retropubic sling passage for the previously operated patient undergoing pubovaginal sling. PMID- 10374791 TI - Diagnostic and therapeutic laparoscopy for the adult cryptorchid testicle. AB - Laparoscopy can be utilized in the management of male patients with impalpable gonads. However, there have been few reports of its application in the management of adult men with cryptorchidism. Fourteen adult men with 15 undescended testicles were referred to our urology department over 6 years. Eight testicles from seven men could not be localized clinically or radiographically to the inguinal canal. Laparoscopy was utilized to assess for the presence and location of the gonad and to perform orchiectomy when testicular tissue was encountered. The average age of the seven men undergoing laparoscopy was 40 years. Three testicles were found proximal to the internal ring and were removed using laparoscopic techniques. The ipsilateral testicular vessels and vas deferens were visualized entering the internal ring in three cases and dense ipsilateral adhesions from previous inguinal surgery prevented adequate visualization of the cord structures in one individual, leading to four inguinal explorations in the same setting. Two inguinal orchiectomies were performed and two vanishing testicles were identified. Gonadal vessels were absent in the remaining patient as demonstrated by extensive laparoscopic dissection. The average total operative time was 105 minutes, which included additional procedures in four patients. No patient experienced an intraoperative or postoperative complication. Six of seven patients were discharged on the day of surgery. An intratesticular germ cell neoplasm was found in one of seven specimens, which stresses the importance of diagnosing and managing this uncommon problem in the adult man. PMID- 10374792 TI - Percutaneous access techniques in renal surgery. AB - Percutaneous renal surgery is continually being improved, refined, and embraced by urologists worldwide. With the advent of extracorporeal shock wave lithotripsy (ESWL), many percutaneous techniques have been abandoned or forgotten. We are learning, however, that ESWL is not a panacea for all urinary calculi and different methods need to be used to obtain stone-free patients. We discuss the history, anatomy, techniques, and specific problems and complications ofpercutaneous renal surgery specifically relating to renal stone disease. PMID- 10374793 TI - Percutaneous cystolithotomy for vesical calculi: a better approach. AB - Vesical calculus is a common problem that is treated traditionally with open cystolithotomy or cystolithalopaxy. Open surgery has the inherent problems of a long scar, prolonged catheterization, extended hospitalization, and risk of infection. Transurethral cystolithalopaxy also requires special instruments that carry a risk of trauma, which could lead to urethral strictures. Thirty-eight patients (15 children and 23 adults) were treated for vesical calculi by percutaneous cystolithotomy (PCCL), a minimally invasive procedure. A fluoroscopic-guided tract was made to the bladder through a small suprapubic puncture (9-10 mm) and a nephroscope was inserted via an Amplatz sheath placed suprapubically. The calculus was fragmented with ultrasound or pneumatic energy before being flushed out. A suprapubic catheter was kept in place for 48 hours postsurgery; no urethral catheter was needed. Urethral instrumentation was kept to a minimum. After 48 hours, the suprapubic catheter was clamped and removed after the patient had two or three normal voids. No significant intraoperative or postoperative complication was encountered. Given that the urethra is spared, percutaneous cystolithotomy is a preferred approach in patients with vesical calculi. PMID- 10374794 TI - Renal malacoplakia. AB - We present a well-documented case of biopsy-proven renal malacoplakia with an excellent response to oral fluoroquinolone therapy. PMID- 10374795 TI - Holmium: YAG laser endoureterotomy in the treatment of ureteroenteric strictures following orthotopic urinary diversion. AB - The management of ureteroenteric strictures in patients who have undergone urinary diversion can be challenging. In those patients with an orthotopic neobladder, anastomotic ureteral strictures can be treated endoscopically using a retrograde or antegrade approach. The availability of small (7.5F) flexible ureteroscopes, as well as the use of the Holmium laser has facilitated the ability to precisely incise the stricture under direct endoscopic visualization (endoureterotomy). We describe our technique for laser endoureterotomy in patients with ureteroenteric strictures following orthotopic urinary diversion. PMID- 10374796 TI - Gunshot wound to the penis: the need for corporal exploration. AB - Although the overall incidence of penetrating penile injuries is relatively low, these injuries may present complex management problems and should be managed with attention to long-term cosmetic and functional considerations. In this article we present an unusual case of a gunshot wound to the penis and emphasize the importance of bilateral corporal exploration and possible repair to avoid erectile dysfunction and penile deformity. PMID- 10374797 TI - Use of talcum in sclerotherapy of pelvic lymphoceles. AB - Lymphoceles sometimes constitute an inconvenient problem after pelvic surgery combined with lymphadenectomy. We report on a case of a patient with carcinoma of the prostate treated by radical prostatectomy and pelvic lymphadenectomy, who developed a large pelvic lymphocele after surgery. The lymphocele was drained by a nephrostomy tube and sclerotherapy using a talcum solution was performed. Talcum is frequently used successfully in pleurodesis. The lymphocele disappeared within 2 weeks after one administration of the solution. Sclerotherapy with a talcum solution can be recommended as a simple, safe, and effective method in pelvic lymphoceles. The single dose and the lack of side effects are advantageous to other agents. PMID- 10374798 TI - Conservative treatment of acute Ormond's disease. AB - Ormond's disease is a chronic inflammatory process of the retroperitoneum, which sometimes takes a very acute course. We report the case of a man with acute bilateral hydronephrosis. Diagnosis was based on typical criteria seen by magnetic resonance imaging, which made biopsy unnecessary. PMID- 10374799 TI - Case no. 1. Bilateral testicular pain. Tubular ectasia of rete testis. PMID- 10374800 TI - Case no. 2. Right testicular pain. Testicular microlithiasis. PMID- 10374801 TI - Case no. 3. Right flank pain. Crixivan lithiasis. PMID- 10374802 TI - Case no. 4. Chronic left flank pain. Ureteral fibroepithelial polyp causing gross hydronephrosis and hydroureter. PMID- 10374803 TI - A study of ureteric peristalsis using a single catheter to record EMG, impedance, and pressure changes. AB - Ureteric peristalsis transports a urinary bolus from the renal pelvis to the bladder. We developed an intraluminal catheter with a pressure transducer on it to study intraluminal pressure changes and a twin bipolar electrode to record the ureteric EMG and impedance (Z) changes during a peristaltic wave. Five female New Yorkshire pigs (50-60 kg) were studied under light halothane anesthesia (5% at induction/1% for maintenance). A steady state of hydration was maintained using intravenous saline infusion. EMG spike burst activity was studied at a 10-cm interval using low (0-30) Hz filters. Impedance between the same electrodes is measured simultaneously in higher frequencies (1-5 KHz) as a function of ureteric motor activity. Pressure generation in the ureteric lumen was also measured simultaneously by a transducer on the same catheter. A digital signal processing program (Poly 4.9) was used for analysis. Parenteral furosemide was used to induce diuresis. Resting ureteric impedance (Z(R)) decreases to Z(B) (Z bolus) during the passage of the urinary bolus. Passage of a contractile zone during a peristaltic wave increases impedance from Z(B) to its Z(R) level and initiates a pressure rise. Bolus length (the length Z(B)) is not constant and decreases distally. EMG corresponds well in time to impedance. Z(R) disappears after infusion of furosemide because of increased urine load and changes of intraluminal ionic environment. The contractile segment of a ureteric peristaltic wave appears to be represented by an elevated Z segment (Z(C)). Pressure rise is recorded only at the beginning of a contractile zone. A specially adapted intraluminal catheter can be used to study peristalsis in the upper urinary tract. One can study all the three components of ureteric peristalsis (excitation, contraction, and intraluminal pressure rise) using such a catheter. PMID- 10374804 TI - Influence of self-care education on illness behaviors and health locus of control of Mexican American women. AB - The purpose of this study was to examine the influence of self-care education on illness behaviors and health locus of control of Mexican American women. Participants were randomly assigned to a control (n = 60) or experimental group (n = 60). Subjects completed the Multidimensional Health Locus of Control questionnaire and an Illness Behavior Assessment at pretest and 6-months. The experimental group received a self-care manual and participated in two 2-hour seminars on how to effectively use the textbook. The experimental group demonstrated a significant increase in self-care behaviors, and significant changes in Internal Health Locus of Control and Powerful Others Health Locus of Control. Chance Health Locus of Control was found to have a low, direct correlation with age, and a low, indirect correlation with education. The conclusion of this investigation is that self-care education can positively influence illness behaviors and Health Locus of Control in Mexican American women. PMID- 10374805 TI - The mental health status of young Hispanic women residing along the border: a twin cities comparison. AB - A number of studies report comparisons among ethnic/ racial groups in terms of health attitudes, health practices, and socio-economic and mental health status. Of special concern is the mental health status and coping potential of young women of childbearing age because of the special vulnerability of individuals in this group and the vulnerability of their children. The well-being of future generations is at stake when maladaptive functioning compounded by severe social conditions create a climate for inadequate growth and development for large numbers in a population, even for short periods of time. This paper reports the results of a study examining self concepts and mental health status of two distinct populations within one ethnic group-young Hispanic women living on the U.S. side of the Texas-Mexico border versus a similar sample of young Hispanic women living on the Mexico side. Within each sample, the never-pregnant versus ever-pregnant adolescents were compared. The young women in both groups reported intense feelings related to emotional distress. The young women in Ciudad Juarez reported somewhat more positive feelings related to recent well-being. The El Paso women reported a less traditionally feminine persona (they felt more aggressive, confident, successful, energetic, and successful), yet they experienced less happiness, hopefulness, and life satisfaction. However, neither group could be described as reporting positive mental status and those women who had been pregnant were no different than their never-pregnant counterparts. Rather, the results signal serious problems throughout the two populations. Health care and social service workers must recognize and be prepared to address intense personal distresses in both of these young, Hispanic-female, border populations. PMID- 10374806 TI - Traditional birth attendants' advice toward breast-feeding, immunization and oral rehydration among mothers in rural Bangladesh. AB - Traditional birth attendants (TBAs) are regarded as essential child health care providers in Bangladesh. A community-based cross-sectional study was completed using questionnaires and interviews to compare trained and untrained TBAs' advice on (1) breast-feeding, (2) immunizations and (3) oral rehydration therapy as an extended part of their maternity care training. Twenty-eight trained TBAs (TTBAs) and 27 corresponding untrained TBAs (UTBAs) in the Dhaka district were interviewed to investigate the effect of their advice on the three outcome variables of maternal health care. Additionally, 276 questionnaires were distributed to the mothers cared for by these TBAs to determine their knowledge of infant-care practices. In-depth interviews with 25 mothers provided additional insight. While TTBAs may have more knowledge and be more willing to disseminate health care information to mothers with new infants than UTBAs, the mother's health practices were independent of the advice provided by the two groups of TBAs. Additionally, the mother's health practices equaled or exceeded expected norms. PMID- 10374807 TI - Asian-Islamic women and breast cancer screening: a socio-cultural analysis. AB - This article explores religious and socio-cultural issues relevant to breast cancer screening practices among older immigrant Asian-Islamic women in the U.S. Some of the Islamic tenets that facilitate breast cancer screening include cleanliness, prevention and individual responsibility in health promotion, diet and eating habits, and exercise, and those that hinder screening practice include gender and modesty considerations and patriarchal marital beliefs. Socio-cultural barriers include patient-physician communication and beliefs about cancer and cancer prevention. Recommendations to increase knowledge and practice of breast cancer screening within a religious and socio-cultural context are provided. PMID- 10374808 TI - Clandestine abortion in Latin America: provider perspectives. AB - This paper presents the results of in-depth interviews with ten clandestine abortion providers in urban Latin America. Three related issues are addressed: how abortion providers come to this line of work; their major difficulties; and their sources of job satisfaction. A variety of paths bring health professionals to the practice of abortion; common elements are a sense of calling, a desire to help women, personal experience with abortion, and a commitment to political change. Providers describe difficulties that include a lack of medical support, the need for secrecy, and threats of violence, extortion, and prosecution. In spite of difficulties, all providers report a great deal of fulfillment in their work, based on their satisfaction in saving women's lives, maintaining supportive relationships with colleagues, and empowering women. PMID- 10374809 TI - Mothers in a new country: the role of culture and communication in Vietnamese, Turkish and Filipino women's experiences of giving birth in Australia. AB - There are few population based studies which explore the views immigrant women have of the maternity care they receive in their new homelands. Three hundred and eighteen Vietnamese, Turkish and Filipino women who gave birth in three major city hospitals in Melbourne, Australia were interviewed about their experiences of maternity care. Outcomes and experiences for women with different levels of English fluency were studied, as were women's needs and preferences for assistance with interpreting. Observance of traditional cultural practices surrounding birth and the impact of not being able to observe such practices on women's experiences of care were also explored. Women in the study not fluent in English experienced problems in communicating with their caregivers and these were reflected in less positive experiences of care. Women were less concerned that caregivers knew little about their cultural practices than they were about care they experienced as unkind, rushed, and unsupportive. Maternity care for immigrant women is only likely to improve when barriers to effective communication are addressed and attention is paid to raising standards of care. PMID- 10374810 TI - An investigation of sociomedical risk factors associated with vaginal fistula in northern Nigeria. AB - In some countries in Africa it is customary for early marriages involving young adolescent girls to be contracted prior to the commencement of their menses. This practice often results in premature pregnancies which in turn leads to devastating physical and social consequences such as vesico vaginal fistula (VVF). VVF is a severely demoralizing and disabling childbirth injury among women who become incontinent as a result of an opening created between the vagina and bladder. A case control study of 50 VVF patients and 50 non-VVF village women was undertaken in Katsina, Nigeria. Statistical analysis showed that VVF patients were smaller in stature, had less education and were of lower socioeconomic status. Also, the analysis showed that both groups of women married and commenced childbearing at an age too early for a safe delivery, thus placing them at risk of VVF. Predictive variables for the condition are: age at marriage, parity, husband's occupation and level of education. PMID- 10374811 TI - Essential fatty acids are mediators of brain biochemistry and cognitive functions. AB - Major advances have been made in understanding the biochemistry of essential fatty acids (FA) and their interactions with metabolic pathways leading to the production of longer and more complex fatty acids and lipids. Less understood are the roles played by FA which are known to affect neurotransmitters, peptides, releasing factors, hormones, and a variety of physiological and cognitive processes. Based on empirical findings we propose that (a) FA exert a controlling function in the modulation of neuronal membrane fluidity, and (b) the critical factor in FA action and efficacy is not absolute level but rather the ratio between various groups of FA. This approach unifies the biochemical and cognitive results obtained from many different and unrelated fields of research. PMID- 10374812 TI - Opposite effects of interferon-gamma and prostaglandin E2 on tumor necrosis factor and interleukin-10 production in microglia: a regulatory loop controlling microglia pro- and anti-inflammatory activities. AB - Following brain injury, microglial cells produce pro- and anti-inflammatory cytokines, such as tumor necrosis factor (TNF) and interleukin-10 (IL-10). IL-10 provides an efficient autocrine mechanism for controlling microglia activation. To elucidate the mechanisms that regulate the cytokine profile of microglia, we examined the effects of several immunomodulators on IL-10 and TNF production by cultured mouse microglia. Lipopolysaccharide (LPS) was the only inducer of IL-10 and TNF gene expression and secretion. The T helper 1-type cytokine interferon gamma (IFN-gamma) induced TNF transcripts, but not TNF secretion, and suppressed LPS-induced IL-10 mRNA and secretion by microglia. Opposite to IFN-gamma, the lipid mediator prostaglandin E2 (PGE2) enhanced the LPS-induced production of IL 10 and inhibited that of TNF. The effects of PGE2 on cytokine gene expression and secretion were antagonized by IFN-gamma. Agents that increase cAMP levels mimicked the action of PGE2 on cytokine secretion, indicating the involvement of cAMP-coupled prostaglandin receptors. In conclusion, IFN-gamma and PGE2, two mediators released at inflammatory sites, differentially regulate the production of a proinflammatory and an anti-inflammatory cytokine in microglia. We suggest that the activity and role of microglia in the damaged CNS could be finely tuned by the local concentration ratio of these mediators. PMID- 10374813 TI - Long-term survival of fetal porcine lateral ganglionic eminence cells in the hippocampus of rats. AB - Embryonic porcine brain tissue from the lateral ganglionic eminence was transplanted into the adult rat hippocampus to determine whether fetal striatal cells could survive, differentiate, and integrate in a heterotopic site. The hippocampus, a common site of epileptic seizure activity, was chosen to determine if fetal striatal cells could supply inhibitory GABAergic neurons that may serve to block seizures. Cells were either implanted with a single deposit using a standard metal cannula or by five smaller disseminated deposits with a glass micropipette. At 20-24 weeks, animals immunosuppressed with cyclosporin showed long-term survival of porcine cells in the adult hippocampus. Analysis by immunohistochemistry and in situ hybridization showed that the grafts contained glial and neuronal cell types, including GABAergic neurons within graft core and networks of porcine neuronal fibers extending from the graft into the host parenchyma. In addition, a marker of porcine presynaptic terminals, synaptobrevin, was abundant within the grafts and was found associated with hippocampal structures and cell layers suggesting functional integration of grafted cells within the host. The survival of xenografts in the hippocampus and potential integration of inhibitory components provides evidence that these grafts may serve as an internal negative feedback mechanism to quench epileptiform activity. PMID- 10374814 TI - Independent mechanisms of potassium clearance by astrocytes in gliotic tissue. AB - The "glial impairment hypothesis" states that astrocytes which change from normal into the reactive type lose their ability to clear extracellular K+, which in turn leads to hyperexcitability in the gliotic tissue. As this hypothesis was never proven or disproven, the question of glial efficiency in K+ clearance in gliotic tissue is still controversial, mainly due to the lack of direct measurements of the intracellular K+ concentration of reactive astrocytes. In order to investigate K+ accumulation by glial cells of gliotic tissue, we used hippocampal slices. Adult rats, previously treated with kainic acid, exhibited loss of neurons and gliosis in the CA1 layer of the hippocampus within 3 days. After this time period, double-barrelled microelectrodes were used to inject Lucifer yellow into cells of the stratum radiatum of the CA1 subfield in 400 microm-thick hippocampal slices. These cells had electrophysiological and morphological characteristics of astrocytes. Most injected cells (70%) were dye coupled to other cells and were glial fibrillary acidic protein (GFAP)-positive (80%). We found, however, that GFAP-positive cells were dye-coupled not only to each other, but also to GFAP-negative cells. In another set of experiments, we investigated the glial membrane potential during reduction of the extracellular Cl-concentration and the use of the Cl- channel blocker 4,4' diisothiocyanostilbene-2,2' disulphonic acid (DIDS). The results suggest that reactive astrocytes have a significant resting Cl- conductance. K+-selective microelectrodes were used to analyze the intracellular glial K+ concentration. When the extracellular K+ concentration was increased from 3.5 mM to 10 mM, the intracellular K+ concentration increased by 23 mM. Experiments in which different ion transport systems were blocked with ouabain and DIDS suggest that this increase is dependent on two mechanisms, which can substitute each other: the Na+, K+-ATPase and passive K+ and anion fluxes. Inhibition of either of the two mechanisms did not block the K+ uptake. If, however, the Na+, K+-ATPase and Cl- channels were inhibited at the same time, the net accumulation of K+ was blocked. It appears, therefore, that astrocytes in the gliotic stratum radiatum of the hippocampal slice have the capacity to limit increases in extracellular K+ that are produced by hyperactive surviving hippocampal neurons by passive mechanisms and hence independently of blood and oxygen supply. PMID- 10374815 TI - Neuroprotection by pigment epithelial-derived factor against glutamate toxicity in developing primary hippocampal neurons. AB - Pigment epithelial-derived factor (PEDF) has been shown to be a survival factor for cerebellar granule neurons. Here we investigated the ability of PEDF to enhance the survival of hippocampal neurons in culture, and to protect these neurons against acute glutamate toxicity. Hippocampal neurons prepared from 1- to 3-day postnatal rat brain were cultured for either 7 or 14 days in vitro (DIV). At 14 DIV, neurons were only slightly protected (13% +/- 4%) against 50 microM glutamate toxicity when treated with 1 microg/ml of PEDF for 3 successive days before glutamate exposure as measured by lactate dehydrogenase (LDH) release. In comparison, basic fibroblast growth factor (bFGF) at 10 ng/ml for the same treatment period protected 58% +/- 8% of the neurons against glutamate. Using quantitative image analysis of digitized micrographs, we found that the average size of neurons in young, developing hippocampal cultures (7 DIV), was greatly decreased by treatment with 50 microM glutamate. Treatment for up to 5 successive days with 1 microg/ml of PEDF before glutamate addition dramatically increased the average hippocampal neuron soma size, compared to cells treated with glutamate alone. Thus, PEDF may promote the growth of hippocampal neurons, and, if added to developing hippocampal neurons, can also protect these cells from subsequent injury, such as the excitotoxicity of glutamate. PMID- 10374817 TI - PC12 and neuro 2a cells have different susceptibilities to acetylcholinesterase amyloid complexes, amyloid25-35 fragment, glutamate, and hydrogen peroxide. AB - This work addresses the differential effects of several oxidative insults on two neuronal cell lines, PC12 and Neuro 2a cells, extensively used as neuronal models in vitro. We measured cellular damage using the cytotoxic assays for MTT reduction and LDH release and found that acetylcholinesterase (AChE)-amyloid-beta peptide (Abeta) complexes, Abeta25-35 fragment, glutamate and H2O2 were over 200 fold more toxic to PC12 than to Neuro 2a cells. 17alpha and 17beta estradiol were able to protect both cell types from damage caused by H2O2 or glutamate. By contrast, other insults not related to oxidative stress, such as those caused by the nonionic detergent Triton X-100 and serum deprivation, induced a similar level of damage in both PC12 and Neuro 2a cells. Considering that the Abeta peptide, H2O2 and glutamate are cellular insults that cause an increase in reactive oxygen species (ROS), the intracellular levels of the antioxidant compound, glutathione were verified. Neuro 2a cells were found to have 4- to 5 fold more glutathione than PC12 cells. Our results suggest that Neuro 2a cells are less susceptible to exposure to AChE-Abeta complexes, Abeta25-35 fragment, glutamate and H2O2 than PC12 cells, due to higher intracellular levels of antioxidant defense factors. PMID- 10374816 TI - Structural and compositional determinants of cortistatin activity. AB - Cortistatin-14 (CST-14) is a putative novel neuropeptide that shares 11 of its 14 residues with somatostatin-14 (SRIF-14), yet its effects on sleep physiology, locomotor behavior and hippocampal function are different from those of somatostatin. We studied the structural basis for cortistatin's distinct biological activities. As with SRIF-14, CST-14 does not show any preferred conformation in solution, as determined by circular dichroism and nuclear magnetic resonance. Synthetic cortistatin analogs were designed and synthesized based on the cyclic structure of octreotide. Biological assays were carried out to determine their binding affinities to five somatostatin receptors (sstl-5) and their ability to produce changes in locomotor activity and to modulate hippocampal physiology and sleep. The results show that the compound with N terminal proline and C-terminal lysine amide exhibits cortistatin-like biological activities, including reduction of population spike amplitudes in the hippocampal CA1 region, decrease in locomotor activity and enhancement of slow-wave sleep 2. These findings suggest that both proline and lysine are necessary for cortistatin binding to its specific receptor. PMID- 10374818 TI - Characterization of alpha(s)-immunoreactive ADP-ribosylated proteins in postmortem human brain. AB - ADP-ribosylation of the stimulatory G protein alpha subunit, alpha(s), has been demonstrated in a number of different mammalian tissues. However, little is known about the occurrence and role of this process in modifying alpha(s) levels/function in human brain. In the present study, endogenous and cholera toxin (CTX)-catalyzed [32P]ADP-ribosylated products were characterized in postmortem human temporal cortex by (1) immunoprecipitation with alpha(s) antisera (RM/1), (2) comparisons of immunoblots and autoradiograms of the [32P]ADP-ribosylated products, and (3) limited protease digestion. Of the three major endogenous [32P]ADP-ribosylated products (48, 45, and 39 kDa) in postmortem brain, the 48-kDa and 45-kDa bands were clearly identified as alpha(s-L) (long isoform) and alpha(s-S) (short isoform), respectively. RM/1 immunoprecipitated the 39-kDa [32P]ADP-ribosylated protein, and overlays of immunoblots and autoradiograms showed that this product corresponded to an alpha(s)-like immunoreactive protein. Furthermore, limited protease digestion of the 39-kDa endogenous [32P]ADP-ribosylated band generated peptide fragments similar to both endogenous and CTX-catalyzed [32P]ADP-ribosylated alpha(s-S). Two major CTX catalyzed [32P]ADP-ribosylated products were also identified as alpha(s-L) (52 kDa) and alpha(s-S) (45 kDa). These findings clearly demonstrate that alpha(s) is a substrate for endogenous and CTX-catalyzed [32P]ADP-ribosylation in postmortem human brain. Furthermore, a lower molecular weight alpha(s)-like immunoreactive protein is also expressed in human brain and is a substrate for endogenous but not CTX-catalyzed [32P]ADP-ribosylation. PMID- 10374819 TI - Function of microglia in organotypic slice cultures. AB - We have examined the functional characteristics of microglia in an environment where the cytoarchitecture of the brain is preserved. Using organotypic slice culture under serum-free conditions, microglia initially demonstrated a rounded morphology but after 10 days in vitro (DIV), microglia in the slice were highly branched. Stimulation of the microglia at 4 DIV with phorbol ester significantly increased the number of cells stained with nitroblue tetrazolium, an indicator of superoxide anion production, compared to non-stimulated conditions. At 10 DIV, superoxide anion production was significantly less than that seen at 4 DIV and no increase in production was seen with phorbol ester stimulation. Phagocytosis of fluorescent latex beads and chemotaxis of microglia in response to zymosan activated serum was also reduced at 10 DIV compared to 4 DIV. These experiments indicate that microglia at 4 DIV in tissue slice culture have functional characteristics that resemble microglia in primary culture. However, prolonged culture of the slices results in a return of the microglia to a ramified and functionally down-regulated state, reminiscent of an "in vivo"-like environment. The organotypic slice culture, thus, provides a useful model system to I examine the interactions of microglia with neurons and other glia in the normal and injured brain. PMID- 10374820 TI - Protective role of heme oxygenase-1 in oxidative stress-induced neuronal injury. AB - Heme oxygenase-1 (HO-1) is a stress protein induced in response to a variety of oxidative challenges. After treatment of the hybrid septal cells SN 56 with beta amyloid peptide (beta-AP1-40) or hydrogen peroxide (H2O2), we detected high levels of reactive oxygen species, accompanied by a significant elevation in HO-1 expression. Levels of HO-1 increased and then decreased following cell loss. Pretreatment of SN 56 cells with HO-1 antisense oligonucleotides dramatically decreased the immunoreactivity of HO-1 and significantly enhanced the cytotoxicity of beta-AP1-40 and H2O2. In contrast, pretreatment with hemin, an HO 1 inducer, increased the expression of HO-1 and decreased the beta-AP1-40- and H2O2-induced cytotoxicity. These findings support the importance of HO-1 in protecting neurons against oxidative stress-induced injury. PMID- 10374821 TI - Activation and inactivation of taurine efflux in hyposmotic and isosmotic swelling in cortical astrocytes: role of ionic strength and cell volume decrease. AB - A decrease in intracellular ionic strength appears involved in the activation of swelling-elicited 3H-taurine efflux in cortical cultured astrocytes. Hyposmotic (50%) or isosmotic urea-induced swelling leading to a decrease of intracellular ionic strength, activated 3H-taurine efflux from a rate constant of about 0.008 min(-1) to 0.33 min(-1) (hyposmotic) and 0.59 min(-1) (urea). This efflux rate was markedly lower (maximal 0.03 min(-1)) in isosmotic swelling caused by K+ accumulation, where there is no decrease in ionic strength, or in cold (10 degrees C) hyposmotic medium (maximal 0.18 min(-1)), where swelling is reduced and consequently intracellular ionic strength is less affected. Also, astrocytes pretreated with hyperosmotic medium, which recover cell volume by ion accumulation, did not release 3H-taurine when they swelled by switching to isosmotic medium, but when volume was recovered by accumulation of urea, taurine release was restored. These results point to a key role of ionic strength in the activation of osmosensitive 3H-taurine efflux. In contrast, its inactivation was independent of the change in ionic strength but appears related to the reduction in cell volume after swelling, since despite the extent or direction of the change in ionic strength, the 3H-taurine efflux did not inactivate in isosmotic KCl-elicited swelling when cell volume did not recover nor in hyposmotic swelling when RVD was impaired by replacing NaCl in the medium by permeant osmolytes. PMID- 10374822 TI - Selective subcellular redistributions of protein kinase C isoforms by chemical hypoxia. AB - The mechanisms of neuronal degeneration following hypoxia/ischemia remain undefined, but the processes include increases in neurotransmitter release, elevation of cytosolic-free calcium concentration, and changes in signal transduction pathways. Activation of the multigene family of protein kinase C (PKC) has been associated with the release of neurotransmitter and the survival of neurons. Therefore, to understand which PKC isozymes are involved in hypoxia/ischemia-induced neuronal degeneration, we examined PKC isozymes after chemical hypoxia (i.e., KCN exposure) in PC12 cells. Cell toxicity, as measured by lactate dehydrogenase (LDH) release, was increased significantly by KCN in glucose-free DMEM and was exaggerated by acute 12-O-tetradecanoyl phorbol-13 acetate (TPA) pretreatment. Under parallel conditions, KCN elevated cytosolic free calcium ([Ca2+]i) in glucose-free but not in glucose containing DMEM, and TPA pretreatment did not exaggerate KCN's effect on [Ca2+]i. Thus, increases in [Ca2+]i are not sufficient for the synergistic toxic effect of KCN and TPA. In the glucose-free DMEM, selective PKC isozyme inhibitor Go 6976 at 10 nM completely inhibited KCN-induced LDH release and at higher concentrations (1 microM) inhibited the basal levels of LDH release. The protein levels of PKCs in the nuclear, membrane, and cytosolic fractions were measured by Western blot analysis using antibodies against specific isoforms. Two Ca2+-dependent (-alpha, gamma) and four Ca2+-independent (-delta, -epsilon, -zeta, and -lambda) isozymes were identified and two isozymes (-beta and -theta) were not detected in the subcellular fractions of PC12 cells. Treatment of the cells with TPA significantly activated translocation of conventional PKC-gamma from the cytosol to the membrane and nuclear fractions and other PKC isozymes (-alpha, -delta, and -epsilon) from the cytosol to the membrane, but not atypical PKC-zeta and lambda. Although only the levels in the nuclear PKC-gamma but not other PKC isozymes were increased significantly following KCN, the levels of cPKC-alpha and -gamma in the membrane mainly- and those and PKC-epsilon in the nucleus-were increased when KCN was combined with TPA. In addition, this condition (TPA + KCN) did not affect the TPA insensitive atypical isozymes, PKC-zeta or -lambda. Taking the results together, differential activation/translocation of PKC isozymes by KCN and TPA is important in the regulation of chemical hypoxia-induced cell injury in PC12 cells. PMID- 10374824 TI - Respiratory resistance measured by an airflow perturbation device. AB - The airflow perturbation device (APD) is an instrument for the measurement of respiratory resistance. The APD is small, lightweight, fast and requires no special breathing manoeuvres. Airflow perturbation determines resistance by superimposing a periodic signal onto spontaneous breathing with a variable resistance device. Respiratory impedance is the ratio of magnitude of pressure perturbation to magnitude of flow perturbation, and respiratory resistance is the in-phase portion of respiratory impedance. The APD was tested to determine its responses to repeated resistance measurements and to changes in resistance. A mechanical model test showed that the APD could detect increased resistance levels, but overestimated resistance when resistance increased with flow. Tests with human subjects showed that the APD gave results consistent from day to day, was able to detect added resistances, and gave resistance values correlated with airway resistance values obtained by body plethysmography. Accelerometers placed on the chests of the subjects showed that perturbations extend to the chest surface. Thus, the APD must measure total respiratory resistance. PMID- 10374823 TI - Tomographic reconstruction of the retina using a confocal scanning laser ophthalmoscope. AB - Retinal imaging with a confocal scanning laser ophthalmoscope (cSLO) involves scanning a small laser beam over the retina and constructing an image from the reflected light. By applying the confocal principle, tomographic images can be produced. However, the thickness of such slices, when compared with the retinal thickness, is too large to give useful 3D retinal images. In this study an algorithm has been developed which fits a double Gaussian curve to the axial intensity profiles generated from a stack of image slices. The underlying assumption is that the laser light has mainly been reflected by two structures in the retina, the internal limiting membrane and the retinal pigment epithelium. From the fitted curve, topographic images and novel thickness images of the retina can be generated. The technique has been applied to three normal volunteers and seven patients with macular pathology (cystoid macular oedema and macular hole) demonstrating the clinical value of the technique. The improvement in accuracy achieved by using a double rather than a single Gaussian is also demonstrated. PMID- 10374826 TI - Evaluation of a non-invasive respiratory monitoring system for sleeping subjects. AB - In a previous paper we introduced a non-invasive respiratory monitoring system (NIRMS) for monitoring respiratory movements during sleep. Unlike standard sleep laboratory methods, the NIRMS can be used in frail older adults to describe breathing patterns during sleep that will mark individuals with declining neurological function. The present study evaluates the use of the NIRMS as a respiratory monitor and identifies variables that can reliably detect changes in breathing patterns and the presence of other body movements. Data were obtained from eleven healthy adults (six women, five men) whose body mass indices ranged from 20 to 47 kg m(-2), and whose baseline respiratory rates ranged from 4 to 19 breaths per minute. We evaluated three variables derived from frequency and amplitude measurements of the NIRMS: (1) the interval between breathing cycles (the interbreath interval or IBI); (2) the period between breathing cycles (the interbreath frequency or IBF); and (3) the amplitude of breath cycles (AMP). The frequency of NIRMS waveform deflections correlated highly with the frequency of visually observed chest movements (r = 0.99). Compared with a subject's baseline, a standard deviation of IBI > 3.0 s consistently identified time segments with three or more apnoeic events. The IBF and AMP differentiated respiratory from other body movements. An IBF > 20 cycles/s or an AMP > 0.4 V identified experimentally introduced body movements much more accurately than wrist accelerometry (NIRMS detected 99% of events, kappa = 0.90; WA detected 50%, kappa = 0.70). These findings support the use of the NIRMS in monitoring changes in breathing patterns during sleep, especially in frail and cognitively impaired subjects. PMID- 10374825 TI - A technique for global assessment of respiratory muscle performance at different lung volumes. AB - A system for noninvasive assessment of an all-inclusive function of respiratory muscles at different lung volumes is presented. The apparatus was based on the interrupter technique and facilitated simultaneous measurements of mouth pressure and airflow rate during dynamic or quasistatic manoeuvres. In this study, mouth pressure values were continuously acquired during and after interruption of a forced inspiratory or expiratory manoeuvre for as long as the subject could sustain an elevated mouth pressure against the obstructed opening. These measurements provided information on both muscle strength and power. A total of 420 forced maximal inspiratory and expiratory manoeuvres performed by six healthy subjects were monitored at different lung volumes. The pattern of maximal pressure-time curves was consistent for the same subject regardless of lung volume. Similar values of maximal mouth pressure can be generated by healthy subjects by using either a flange-style mouthpiece or facial mask. For both methods mouth pressure shows a significant (p < 0.05) second order dependency on lung volume for both inspiration and expiration. The standard deviation of measurements from a single subject about a second order curve is of the order of 5-15%. The findings of interchangeability between methods of measurement may be useful in allegedly non-compliant patients. PMID- 10374827 TI - Combined microlightguide spectrophotometry and microendoscopy for measurement of oxygen saturation in peripheral nerves. AB - Microlightguide measurements of the spectral composition of backscattered light may be used to determine local tissue oxygen saturation and monitor tissue perfusion using intravenous injection of fluorescein dye as a contrast agent. We have used a combination of microlightguide spectrophotometry and microendoscopy to measure intravascular oxygen saturation (HbSaO2%) and monitor blood flow in the sciatic nerve of 12 healthy male Sprague-Dawley rats. The microlightguide and endoscope combination is a relatively new measurement technique. The aims of this study were to determine whether microlightguide spectrophotometry and microendoscopy could be used to measure HbO2 and blood flow in peripheral nerves and to compare the measurements made using the flexible lightguide with the endoscope-lightguide combination. We found no significant difference between the two types of measurement over similar regions of the nerve. mean SaO2% values 77.1% (95% CI = 75.4-78.8) and 78.8% (95% CI = 77.5-80.1) respectively. During a period of hypoxia there was a similar fall in both arterial and nerve oxygen saturation. Following injection of fluorescein, the rate of increase in nerve fluorescence was used as a measure of perfusion. The combination of microlightguide spectrophotometry and microendoscopy allows the exact site of measurement to be directly visualized. The minimally invasive nature of this technique may allow its application to the study of peripheral nerves in human subjects in conditions such as diabetic neuropathy where vascular factors are thought to have an important role in aetiology. PMID- 10374828 TI - Cerebral blood volume measurements using dynamic contrast-enhanced x-ray computed tomography: application to isoflurane anaesthetic studies. AB - A convenient and simple method of measuring cerebral blood volume (CBV) will aid in the clinical management of patients with cerebrovascular diseases and head trauma. Using a two-compartment model to characterize the distribution of x-ray contrast agent in the brain, we have developed a non-equilibrium (dynamic) x-ray CT method to measure CBV and have applied this method to study the effects of isoflurane. CBV and cerebral blood flow (CBF, ex vivo) measurements were made in two groups of New Zealand White rabbits at varying (study group) and constant (control group) arterial CO2 concentration. ANOVA for repeated measures was used on the control data to determine the precision of our two-compartment CT-CBV method. The results showed that the precision of our CT-CBV measurement was 9.8%. In addition a paired t-test analysis of the control data revealed that the duration of isoflurane anaesthesia had no effect on the repeated measurements of CBV or CBF. The rate of change in CBV (0.049 ml/100 g/mmHg) and CBF (6.2 ml/min/100 g/mmHg) with respect to changes in arterial CO2 concentration under the influence of isoflurane anaesthesia was similar to those reported in previous studies, thus validating our experimental CT-CBV and CBF measurements. Our CT-CBV method will lead to more information on the relationship between CBV and CBF under different pharmacological interventions in both normal and disease conditions. PMID- 10374829 TI - Incorporating a priori anatomical information into image reconstruction in electrical impedance tomography. AB - Image reconstruction in electrical impedance tomography using the sensitivity theorem is generally based on the assumption that the initial conductivity distribution of the body being imaged is uniform. The technique of image reconstruction using this method is described and reconstructed images are presented. Improvements in image quality and accuracy are demonstrated when accurate a priori 'anatomical' information, in the form of a model of the distribution of conductivity within the region to be imaged, is used to construct the sensitivity matrix. In practice correct a priori information is not available, for example the conductivity values within the various anatomical regions will not be known. An iterative algorithm is presented which allows the conductivity parameters of the a priori model to be determined during reconstruction. PMID- 10374830 TI - Real time impedance plots with arbitrary frequency components. AB - Temporal changes in the impedance spectra of bioelectrodes in contact with skin provide useful data for comparisons between differing electrode materials and skin preparation methods. Traditional impedance measuring systems employ swept frequency methods which will distort results in the impedance spectra of a sample which has nonstationary electrical characteristics. The system reported here obtains impedance spectra by applying a digitally constructed waveform consisting of many frequency components. This allows impedance values to be measured at a number of frequencies simultaneously. Sample data are provided and compared with data obtained using similar square wave techniques. PMID- 10374831 TI - Nonlinear curve fitting for bioelectrical impedance data analysis: a minimum ellipsoid volume method. AB - Bioelectrical impedance spectroscopy has become a widely used method with various applications in physiological and clinical investigations. Its measurement results are often analysed using the Cole-Cole model in an attempt to relate its parameters to structural or physiological characteristics of the studied object. The model parameter values are usually assessed by nonlinear curve fitting based on the least mean square method. A new approach is proposed, leading to better accuracy. It is based on the notion that the difference between measured and model parameter values is randomly distributed with zero mean. The values of the unknown parameters are then deduced by minimizing the covariance matrix determinant. This method can be considered as a minimization of the volume of an ellipsoid, or ajoint minimization of two regressions having the same argument and common parameters. PMID- 10374832 TI - Evaluation of butylated hydroxyanisole, myo-inositol, curcumin, esculetin, resveratrol and lycopene as inhibitors of benzo[a]pyrene plus 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanone-induced lung tumorigenesis in A/J mice. AB - The potential activities of butylated hydroxyanisole (BHA), myo-inositol, curcumin, esculetin, resveratrol and lycopene-enriched tomato oleoresin (LTO) as chemopreventive agents against lung tumor induction in A/J mice by the tobacco smoke carcinogens benzo[a]pyrene (BaP) and 4-(methyl-nitrosamino)-1-(3-pyridyl)-1 butanone (NNK) were evaluated. Groups of 20 A/J mice were treated weekly by gavage with a mixture of BaP and NNK (3 micromol each) for 8 weeks, then sacrificed 26 weeks after the first carcinogen treatment. Mice treated with BHA (20 or 40 micromol) by gavage 2 h before each dose of BaP and NNK had significantly reduced lung tumor multiplicity. Treatment with BHA (20 or 40 micromol) by gavage weekly or with dietary BHA (2000 ppm), curcumin (2000 ppm) or resveratrol (500 ppm) from 1 week after carcinogen treatment until termination had no effect on lung tumor multiplicity. Treatment with dietary myo-inositol (30,000 ppm) or esculetin (2000 ppm) from 1 week after carcinogen treatment until termination significantly reduced lung tumor multiplicity, with the effect of myo inositol being significantly greater than that of esculetin. Treatment with dietary LTO (167, 1667 or 8333 ppm) from 1 week before carcinogen treatment until termination had no effect on lung tumor multiplicity. The results of this study demonstrate that BHA is an effective inhibitor of BaP plus NNK-induced lung tumorigenesis in A/J mice when administered during the period of carcinogen treatment and that, among the compounds tested, myo-inositol is most effective after carcinogen treatment. PMID- 10374833 TI - Inhibition by D-limonene of gastric carcinogenesis induced by N-methyl-N'-nitro-N nitrosoguanidine in Wistar rats. AB - The effect of prolonged oral administration of D-limonene on gastric carcinogenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and on the labeling and apoptotic indices of gastric cancers were investigated in Wistar rats. After 25 weeks of the carcinogen treatment, rats were given chow pellets containing 1% or 2% limonene. In week 52, long-term oral administration of 2%, but not 1%, limonene significantly decreased the incidence of gastric cancers. Limonene also significantly decreased the labeling index and significantly increased the apoptotic index of gastric cancers. No K-ras mutations were detected in gastric cancers induced by MNNG in either group. These findings indicate that limonene inhibits the development of gastric cancers through increased apoptosis and decreased DNA synthesis of gastric cancers, but not through ras oncoprotein plasma membrane association. PMID- 10374834 TI - Tumorigenicity and liver tumor ras-protooncogene mutations in CD-1 mice treated neonatally with 1- and 3-nitrobenzo[a]pyrene and their trans-7,8-dihydrodiol and aminobenzo[a]pyrene metabolites. AB - The environmental pollutants 1- and 3-nitrobenzo[a]pyrene (1- and 3-NBaP) are metabolized by mammalian microsomes through ring oxidation to 1-NBaP trans-7,8 dihydrodiol and 3-NBaP trans-7,8-dihydrodiol, and by nitroreduction to 1- and 3 aminobenzo[a]pyrene. To determine if these compounds are tumorigenic, 1- and 3 NBaP, along with several of their metabolites and the parent benzo[a]pyrene (BaP) and its trans-7,8-dihydrodiol metabolite, were tested in the neonatal CD-1 mouse bioassay. Male mice were administered i.p. injections at a total dose of 100 or 400 nmol per mouse on 1, 8 and 15 days after birth. While the liver tumor incidences for BaP, BaP trans-7,8-dihydrodiol, and the positive control 6 nitrochrysene (6-NC) were significantly higher than in the solvent control animals, all the other tested compounds exhibited no tumorigenicity. The frequency of Ha- and Ki-ras mutations in liver tumors of mice treated with BaP, BaP trans-7,8-dihydrodiol, and 6-NC were higher than in the few liver tumors isolated from control mice or mice treated with the NBaPs or their metabolites. Since 1- and 3-NBaP and their metabolites are potent mutagens in the Salmonella assay and moderate mutagens in the Chinese hamster ovary (CHO) mammalian mutagenicity assay, our results indicate that the in vitro mutagenicity of these compounds does not correlate with their tumorigenicity. PMID- 10374835 TI - Angiogenesis in ductal breast carcinoma. Comparison of microvessel density between primary tumour and lymph node metastasis. AB - Angiogenesis has been recognised as an important prognostic factor in cancer. This study assessed immunohistochemically (JC70 MoAb) the microvessel score (MS; in x 250 fields) in 35 breast ductal carcinomas with lymph node involvement. Sections from both primary tumours and invaded nodes were assessed. A significantly lower MS was observed in the metastatic foci (24.6 +/- 9 vs. 13.6 +/- 6; P < 0.0001) than in the primary lesions. However, linear regression analysis showed a significant direct correlation between the MS assessed in primary lesions and the related metastatic foci in the lymph nodes (P = 0.006, r = 045). The primary tumour to node (T/N) ratio was assessed; 11/35 cases had a node MS close to the score of the primary (T/N < 2), whilst 17/35 had a T/N ratio ranging from 3 to 7. Extracapsular node involvement was more frequent in cases with low T/N ratio. Angiogenesis in the metastatic foci was independent of the amount of growing stroma within the metastasis. Vessel density in the normal lymph nodes did not correlate with the MS within the node metastases. We conclude that cancer cells migrating to the nodes may have similar angiogenic abilities to the parental cells of the primary tumour. However, environmental factors probably related to node immune reaction against the invading tumour could be responsible for the reduced angiogenicity in the nodes. Further studies are required to investigate the suggested angiogenesis suppressing immune mechanisms occurring in the invaded lymph nodes of breast cancer patients and possible clinical implications. PMID- 10374836 TI - Humoral immune response to p21WAF1/CIP1 in tumor patients, non-tumorous patients and healthy blood donors. AB - We performed a serological analysis for anti-p21WAF1/CIP1 antibodies in sera of patients with different gynecological diseases such as breast cancer, ovarian carcinoma, cervix carcinoma and benign gynecological tissue alterations and from healthy blood donors using the immunoblotting technique with recombinant p21WAF1/CIP1 as antigen as well as a newly designed ELISA. We detected antibodies specific for p21WAF1/CIP1 in sera derived from cancer patients, as well as from patients with non-malignant diseases and from healthy blood donors. Thus, the presence of antibodies against p21WAF1/CIP1 is not a marker for malignancies. Some of the sera with antibodies against p21WAF1/CIP1 also contained antibodies against the oncoprotein mdm2, and/or the growth suppressor gene product p53. The presence of antibodies against p53 correlates with a malignant disease. PMID- 10374837 TI - Enhanced Sp1 DNA-binding activity in murine keratinocyte cell lines and epidermal tumors. AB - Altered regulation of ornithine decarboxylase (ODC) is frequently observed in epidermal tumors. We have shown that the transcription factor Sp1 is one of the regulators of ODC expression and that Sp3 antagonizes this Sp1-mediated activation of ODC expression. These results led us to examine the levels and binding activity of Sp1 and Sp3 in nuclear extracts prepared from cultured murine keratinocytes, transformed keratinocyte cell lines and epidermal tumors. Here we show that the Sp1 DNA-binding activity is higher in established keratinocyte cell line extracts than in primary keratinocyte extracts. Sp1 message levels and Sp1 DNA-binding activity was found to be low in 20-week papillomas and high in squamous cell carcinomas. These results suggest that increased levels of Sp1 and enhanced Sp1 DNA binding activity are correlated with epidermal tumor progression. Based on these results, we propose that increased Sp1 DNA binding may augment the proliferative capacity of tumor cells through overexpression of Sp1-responsive genes, possibly including ODC. PMID- 10374838 TI - Growth inhibition of neuroblastoma cells by lovastatin and L-ascorbic acid is based on different mechanisms. AB - Hydroxymethyl-glutaryl-CoA-reductase (HMG-CoA-reductase), the key enzyme for cholesterol synthesis and essential for the synthesis of the precursor for p21ras farnesylation, was inhibited in neuroblastoma cells by lovastatin or L-ascorbic acid. Both compounds inhibited clonogenic colony formation of neuroblastoma cells in soft agar. However, while the addition of mevalonate, the product of HMG-CoA reductase, circumvented the inhibition by lovastatin it had no reversing effect on the inhibition by L-ascorbic acid. The role of reactive oxygen compounds generated by the degradation of catecholamines, and the pro-oxidative effects of L-ascorbic acid are discussed as mechanisms of action of L-ascorbic acid. PMID- 10374839 TI - Mutations associated with in vivo aflatoxin B1-induced carcinogenesis need not be present in the in vitro transformations by this toxin. AB - Ingestion of aflatoxin B1 is implicated in the high incidence of human liver cancers in several developing countries. An association has been detected between human exposure to aflatoxins, and mutations in the third base of codon 249 of the p53 gene in hepatomas. In vitro experiments using human cell line cells and aflatoxin B1 have demonstrated the induction of p53 mutations in codon 249 and adjacent codons. It was therefore of interest to see if this correlation between the in vivo and in vitro situations held for other species. The present study examined a rat liver-derived cell line, transformed in vitro with aflatoxin B1, for the presence of mutations associated with in vivo aflatoxin-induced hepatocarcinogenesis. In an in vivo rodent model systems using the aflatoxin B1 sensitive male F344 rat, previous studies have shown that hepatocarcinogenesis is accompanied by significant incidences of codon 12 mutations in K-ras and codon 13 mutations in N-ras genes, but in contrast to the human, apparently not by mutations in codon 243 of the p53 gene (which corresponds to codon 249 in the human gene). In contrast to the situation in humans, mutation in the third base of codon 243 in the rat would not result in any changes in amino acid sequence, but mutations in codon 250, as seen in in vitro human systems, would be expressed in the rat p53 protein. In the present study, an immortalised, non-transformed liver epithelial cell line derived from a male F344 rat was transformed in vitro by aflatoxin B1 as demonstrated by tumour formation in nude mice. The transformation was dependent on metabolic activation of the aflatoxin B1. Transfection of DNA, extracted from these tumours, into NIH 3T3 fibroblasts conferred a stable, malignant transforming capacity. However, no mutations in codon 12 of the K-ras or codon 13 of the N-ras genes were detected in any of these tumours. These results indicate that in vitro transformation does not necessarily involve the same mutations, as those observed in vivo. Also, no mutations in codon 243 or adjacent codons of the p53 gene, paralleling those observed in the human cell line treated with aflatoxin B1, were detected. The results serve to emphasise the in vivo and in vitro variation in the oncogene activation in the same target organ or cell lines derived from that organ, even when using a single carcinogen activated by a known metabolic pathway. PMID- 10374840 TI - Human p53(264-272) HLA-A2 binding peptide is an immunodominant epitope in DNA immunized HLA-A2 transgenic mice. AB - C57BL/10 mice transgenic for HLA-A2 were immunized with either a full-length DNA construct of the tumor suppressor p53 or with a minigene encoding the p53-derived immunodominant peptide p53(264)LLGRNSFEV272 (L9V). Vaccination with the full length p53 construct induced potent cytotoxic activity of splenocytes against L9V pulsed target cells after in vivo re-stimulation. Vaccination with the L9V encoding minigene likewise induced specific anti-L9V cytotoxicity in vitro. Subsequent experiments revealed that peptide-pulsed dendritic cells were the most efficient cell types for in vitro re-stimulation. In concordance with this, immunization with L9V-pulsed dendritic cells also induced a potent and specific anti-L9V cytotoxic response in vitro. These data show that HLA-A2/peptide specific cytotoxic immunity can be generated in vivo against the same immunodominant epitope by immunizing either with full-length DNA or with a DNA minigene encoding the immunodominant peptide epitope. PMID- 10374841 TI - Enhanced quinone reductase (QR) activity correlates with promotion potential of diethyl maleate (DEM) in rat forestomach and glandular stomach carcinogenesis initiated with N-methyl-N'-nitrosoguanidine (MNNG). AB - The modifying effect of diethyl maleate (DEM) on gastric tumor development was studied in rats initially given N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and hypertonic sodium chloride (H-NaCl 10% or 5%). Groups of animals were maintained with or without a 0.2% DEM dietary supplement after treatment with MNNG and H NaCl and sacrificed at week 20. Forestomachs and livers cytosolic NAD(P)H:quinone acceptor oxidoreductase (QR) activity was also analyzed. The incidences of forestomach severe hyperplasias in the MNNG + H-NaCl --> DEM groups were also significantly higher than in the MNNG + H-NaCl alone group (P < 0.01 and P < 0.05 for 5% and 10% groups, respectively). Similarly, in the glandular stomach, the numbers of preneoplastic pepsinogen 1 altered pyloric glands (PAPGs) in the MNNG + H-NaCl --> DEM groups were significantly increased (P < 0.01 for both concentrations). The QR activities in the groups treated with DEM showed 2- to 3 fold increases as compared with the control level. The results indicate that treatment with 0.2% DEM after MNNG initiation exerts enhancing effects on both forestomach and glandular stomach carcinogenesis. Induction of QR, a Phase II enzyme, activity in the rat stomach by DEM may be associated with promotion of stomach carcinogenesis rather than inhibition. PMID- 10374842 TI - Expression of highly polysialylated neural cell adhesion molecule in pancreatic cancer neural invasive lesion. AB - Neurotropism of pancreatic cancer is one of the hypotheses explaining neural invasion, which is one of the characteristics of pancreatic cancer. In these studies, we immunohistochemically examined neural cell adhesion molecules (NCAM), homophilic adhesion molecules expressed on the nerve cells, as a factor of neurotropism, in 15 pancreatic cancer operatively obtained, especially in neural invasive lesions. We also investigated the role of polysialic acid (PSA), which is attached to NCAM and related to the malignant potential of cancers. NCAM was detected in 66.7% of pancreatic cancers, and in all 9 cases with massive perineural invasion. In neural invasive lesions, however, there were perineurium and endoneurium, which do not express NCAM, between the cancer and nerve cells. PSA was also detected in the pancreatic cancers expressing NCAM. Moreover, PSA expression was stronger in the perineural invasive lesions than in the main tumor and was related to the cancer cell proliferation investigated by Ki-67 staining. It is unlikely therefore, that NCAM plays an important role in neurotropism. However, the NCAM expressed on the pancreatic cancer was attached to PSA, which itself plays an important role in the malignant potential of this disease. PMID- 10374843 TI - Overexpression of matrix metalloproteinase (MMP)-9 correlates with metastatic potency of spontaneous and 4-hydroxyaminoquinoline 1-oxide (4-HAQO)-induced transplantable osteosarcomas in rats. AB - In the present experiment, the expression of matrix metalloproteinase (MMP)-2 and MMP-9, key proteins in the MMP family, and the tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2, antagonistic proteins against MMP-2 and MMP-9, respectively, were investigated by Northern blot analysis in rat transplantable osteosarcomas with high and low metastatic potencies. Two transplantable osteosarcomas, one induced with the carcinogen, 4 hydroxyaminoquinoline 1-oxide (4-HAQO) (COS, chemical carcinogen-induced osteosarcoma), and the other, a spontaneous lesion (SOS, spontaneous osteosarcoma), were repeatedly transplanted from lung nodules to generate lines with high metastatic potency, C-SLM (chemical carcinogen-induced osteosarcoma, selected lung metastatic lesions) and S-SLM (spontaneous osteosarcoma, selected lung metastatic lesions), respectively. MMP-9 was overexpressed in both S-SLM and C-SLM, and TIMP-2 in the case of S-SLM. Neither MMP-2 nor TIMP-1 was overexpressed in either of the transplantable osteosarcomas with high metastatic potentials. The active form MMP-9, studied by zymography, increased in S-SLM and C-SLM but not in SOS and COS. MMP-9 mRNA expression was highly correlated with the gelatinolytic activity of active form MMP-9 (r = 0.85, P < 0.0001) and with the activation ratio of MMP-9 (r = 0.83, P < 0.0001). However, the active form MMP-2 was not detectable in all cases. These results suggest that overexpression of MMP-9 mRNA is one of the essential factors in the acquisition of metastatic potential in rat transplantable osteosarcomas. PMID- 10374845 TI - In reply to an article published in your journal Cancer Letters, 128 (1998) 87 92, by V.N. Bilim et al. PMID- 10374844 TI - The RARgamma selective agonist CD437 inhibits gastric cell growth through the mechanism of apoptosis. AB - Retinoids are differentiation-inducing agents that exhibit multiple functions. Their activities are mediated through interaction with nuclear retinoic acid receptors (RAR) and retinoid X receptors (RXR). We have investigated the activities of synthetic retinoids on the growth of five gastric cancer cell lines. The effects of agonists selective for RARalpha, RARbeta and RARgamma (AM580, CD2019 and CD437, respectively) on cell growth were determined, in comparison to all-trans retinoic acid, by measuring total cellular DNA. AM580 and CD2019 had little or no effect on the growth of all five cell lines. In contrast, the RARgamma agonist CD437 inhibited cell growth up to 90-99% in both retinoic acid sensitive and resistant gastric cancer cells at a concentration of 1 microM. The growth suppression caused by CD437 was accompanied by the induction of apoptosis as judged by morphological criteria and DNA ladder formation. However, the extent of CD437-induced growth suppression was not correlated with RARgamma mRNA levels, which indicates that CD437 induces apoptosis in gastric cancer cells via an RARgamma independent pathway. PMID- 10374846 TI - Diphtheria toxin fused to granulocyte-macrophage colony-stimulating factor and Ara-C exert synergistic toxicity against human AML HL-60 cells. AB - Human granulocyte-macrophage colony-stimulating factor fused to truncated diphtheria toxin (DT388-GM-CSF) sensitized wild-type and Bcl2-overexpressing HL60 human leukemia cells to intoxication by Ara-C based on proliferation and clonogenic assays. The toxin/drug combination showed dramatic synergistic toxicity with combination indices of < 0.1. Synergy was not seen with two other protein synthesis inhibiting drugs--ricin and cycloheximide nor with GMCSF alone. No changes in Ara-C incorporation into cellular DNA or cell cycle occupancy were seen. As compared to exposure to DT388-GM-CSF or Ara-C alone, co-treatment produced significant increases in cytosolic accumulation of cytochrome c, a higher percentage of cells with loss of mitochondrial membrane potential and an increase in reactive oxygen species and morphologic changes of apoptosis, and a greater induction of poly(ADP-ribose) polymerase (PARP) and DNA fragmentation factor 45 (DFF45) cleavage activities of caspase 3. Co-treatment did not significantly alter Bcl2, Bcl-xL, Bax or Fas receptor (FasR), but modestly increased Fas ligand (FasL) protein. These finding suggest that co-treatment with DT388-GM-CSF may lead to a lowered apoptotic threshold and clonogenic survival of human AML blasts due to Ara-C. These observations also suggest that clinical trials of combination therapy may be warranted in patients with AML. PMID- 10374847 TI - Idarubicin overcomes P-glycoprotein-related multidrug resistance: comparison with doxorubicin and daunorubicin in human multiple myeloma cell lines. AB - The clinical utility of anthracyclines like doxorubicin (DOX) and daunorubicin (DNR) for treatment of multiple myeloma (MM) is limited by the occurrence of multidrug resistance (MDR). Highly lipophilic anthracyclines like idarubicin (IDA) might circumvent MDR and thereby enhance chemotherapeutic efficacy. To determine the efficacy of IDA in myeloma cells, the pharmacokinetics and cytotoxicity of IDA and its major metabolite idarubicinol (IDAol) were compared with those of DNR, DOX, and doxorubicinol (DOXol) in the cell line RPMI 8226-S and two MDR sublines (8226-R7 and 8226-Dox40) that overexpress the drug transporter P-glycoprotein (Pgp). Cytotoxicity assays using MTT (viability) or annexin V (apoptosis) showed a 10-50-fold higher potency of IDA compared with DNR or DOX in the MDR variant cell lines. The difference in cytotoxicity was lower in the sensitive parental cell line (3-fold). These results are explained by a better intracellular uptake of IDA compared to DNR in resistant 8226 cell lines. The Pgp-inhibitor verapamil affected IDA uptake only in the most resistant cell line 8226-Dox40. This indicates that IDA is less sensitive than DNR to transport mediated MDR. IDAol was at least 32-fold more cytotoxic than DOXol, and more susceptible to Pgp transport than IDA. These studies demonstrate that the efficacy of IDA in MDR MM cell lines is superior to that of DOX or DNR, and that IDA may become an important drug in the treatment of MM, especially in refractory disease. PMID- 10374848 TI - Autoimmune disease induced by dendritic cell immunization against leukemia. AB - Induction of an optimal immune response will likely be a prerequisite for successful immunotherapy of human leukemias and other malignancies. Dendritic cells are highly effective at inducing an immune response to antigens to which the host is unresponsive, while transgenic expression of the costimulator molecule CD40 ligand (gp39/CD154) and the T cell growth factor interleukin 2 (IL2) are also able to augment immune responsiveness. We therefore investigated whether a combination of these two distinctive approaches to immunostimulation could safely increase the anti-tumor immune response compared to each stimulus alone. We injected BALB/CBYJ mice with syngeneic dendritic cells (DC) exposed to A20 lymphoblastic leukemia cell-derived peptides and proteins which had been acid eluted from the cell surface. In additional mice, the pulsed DC were mixed with genetically modified syngeneic fibroblasts that were expressing CD40 ligand or secreting interleukin 2 (IL2). Three days after their third, weekly, vaccination, they were challenged with parental A20 cells. Tumor growth was suppressed by responses to pulsed DC alone (P < 0.02). This suppression was further enhanced when pulsed DC were coinjected with fibroblasts expressing CD40 ligand and IL2 (P < 0.0005 compared to DC alone) even though CD40 ligand and IL2-expressing fibroblasts alone offered no significant protection in this model. Mice receiving the full complement of immunostimulants either failed to develop visible tumors or developed small tumors which quickly necrosed and regressed, allowing the mice to become long term tumor-free survivors. Antibody mediated depletion of either CD4+ or CD8+ T-cell subset significantly reduced the level of protection afforded by the vaccination. However, it became evident that this intensive stimulation of the immune system lead not only to tumor eradication but also to destruction of cells bearing normal self antigens. Hence, 60 days after challenge with A20 cells all mice in the DC/IL2/CD40 ligand group developed a severe, systemic autoimmune disorder that resembled graft versus host disease and manifest itself by significant peripheral blood cytotoxicity against autologous fibroblasts, blood dyscrasias, gross hepatosplenomegaly, cachexia and fur loss. This phenomenon depended on CD8+ cytotoxic T lymphocytes. Our results therefore suggest that the most effective strategies of immunotherapy against leukemia may also exceed the threshold of anergic cells, leading to a loss of self tolerance to normal self antigens and the induction of an CD8+ anti-self effector response. PMID- 10374849 TI - Establishment and characterization of a novel ALL-L3 cell line (BALM-18): induction of apoptosis by anti-IgM and inhibition of apoptosis by bone marrow stroma cells. AB - A human acute lymphoblastic leukemia (ALL) cell line, BALM-18, was established from the peripheral blood specimen of a patient with B cell ALL L3 type (ALL-L3) at diagnosis using bone marrow stroma cells (BST) as feeder cells. The primary leukemia cells did not grow without feeder cells. As with the primary leukemia cells, BALM-18 showed an immunophenotype of Burkitt's lymphoma group I [CD10+, CD20+, CD23-, CD39-, CD77+] and carried the t(8;14)(q24;q32) chromosomal abnormality which is highly associated with ALL-L3 and Burkitt's lymphoma. It also revealed a significantly low level of bcl-2 protein. Strikingly, anti-human IgM antibody did induce apoptosis in induction experiments. However, it was reversed by the addition of anti-CD40 antibody or BST cells, whereas the culture supernatant of the stroma cells did not show any effect on the inhibition of apoptosis. BALM-18 may be useful for analyzing both the mechanisms of anti-IgM induced apoptosis and signaling during the inhibition of apoptosis by CD40 or BST cells. PMID- 10374850 TI - Paclitaxel-induced apoptosis in Jurkat, a leukemic T cell line, is enhanced by ceramide. AB - We hypothesized that the lipid second messenger, ceramide, and microtubule directed chemotherapeutic agents might engage converging pathways in inducing apoptosis. Our studies demonstrated that simultaneous treatment of Jurkat cells with paclitaxel and ceramide enhanced paclitaxel-induced cell growth inhibition. Cell cycle analysis indicated a significant increase in the hypodiploid population over that observed with paclitaxel treatment alone. Morphologic evaluation and a TUNEL assay confirmed a dramatic increase in apoptosis in Jurkat cells treated with the combination of these two agents. This is the first demonstration that paclitaxel and ceramide interact in a supra-additive manner to decrease leukemic T-cell growth, suggesting a possible application of paclitaxel and ceramide in combination therapy. PMID- 10374851 TI - Neutrophil dysplasia is not a specific feature of the abnormal chromosomal clone in myelodysplastic syndromes. AB - In order to study if dysplastic cells are part of the abnormal chromosomal clone in myelodysplastic syndromes (MDS) we used fluorescence in situ hybridization in combination with standard May-Grunwald-Giemsa morphology (MGG/FISH) to investigate the penetration of chromosomal abnormalities into dysplastic neutrophil granulocytes in five MDS patients with documented monosomy 7 or trisomy 8 in the bone marrow at diagnosis. Neutrophil dysplasia was defined as neutrophil granulocytes with extreme hypogranulation or with nuclear abnormalities of the pelgeroid type. In one patient all dysplastic cells were derived from the abnormal chromosomal clone, while in the other four cases only 6 43% of the hypogranulated neutrophils and 33-40% of the pelgeroid granulocytes exhibited the chromosomal marker. The results suggest that neutrophil dysplasia is not a specific feature of the abnormal chromosomal clone in MDS. It is not clear, however, if the disomic dysplastic cells were derived from a parallel MDS clone with a normal karyotype, or represent residual non-clonal, normal hematopoietic cells. PMID- 10374852 TI - Adult de novo acute myeloid leukemias with MLL rearrangements. AB - MLL gene rearrangements identify a subgroup of acute leukemias with a bad prognosis. Fifty-one adult patients with de novo AML were screened for MLL rearrangements by Southern blot. An unexpected high frequency of AML-M2 according to the FAB classification was found. The most striking morphologic features of these cases were the scarcity of Auer rods and granulocytic dysplasia. Morphologic traits could be helpful in distinguishing molecular groups of AML-M2. PMID- 10374853 TI - Is there still a place for clinical staging in chronic lymphocytic leukaemia? PMID- 10374854 TI - Acute lymphoblastic leukemia with multiple cytogenetic abnormalities secondary to treatment of Ewing's sarcoma. AB - We report the case of a 22-year-old man with Ewing's sarcoma who attained a complete remission (CR) after combination radiotherapy and chemotherapy. Secondary acute lymphoblastic leukemia with multiple cytogenetic abnormalities involving chromosome 5 and 7 developed 16 years later. The patient underwent induction chemotherapy and entered a CR. Peripheral blood stem cell transplantation from a matched sibling was performed successfully and he is in complete remission of both ALL and Ewing's sarcoma. PMID- 10374855 TI - Prevalence of hemochromatosis related HFE gene mutations in patients with acute myeloid leukemia. PMID- 10374857 TI - Development of a simple, blood based lymphoproliferation assay to assess the clinical status of patients with acute lymphoblastic leukemia. AB - Although childhood acute lymphoblastic leukemia (ALL) is highly responsive to chemotherapy, reliable techniques are needed to determine treatment outcome and predict relapse. Employing a 9-O-acetyl sialic acid binding lectin, ATN(H), we have identified two 9-O-acetylated sialogycoconjugates (9-OAcSGs) as novel biomarkers expressed selectively on leukemic blasts of ALL patients. Presently, we report a non-invasive, blood based lymphoproliferation assay, which employs the maximal lymphoproliferative dose of ATN(H) (MLD) to assess the status of 9 OAcSGs with progressive therapy. A low MLD (0.18 +/- 0.01 microg) in untreated patients reflects increased expression of 9-OAcSGs which decline following therapy (MLD = 2.10 +/- 0.60 microg), persist during maintenance therapy (MLD = 4.50 +/- 1.60 microg)/follow-up (MLD = 5.50 +/- 0.85 microg) and are re-induced with relapse (MLD = 0.25 +/- 0.01 microg). Since the assay detects lymphoblasts with a sensitivity of 10(-4), shows no cross-reactivity with other hematological disorders (n = 48) and has been tested in 212 patients, it meets clinical consideration. PMID- 10374856 TI - Comparisons of prognostic scoring systems for myelodysplastic syndromes: a Korean multicenter study. AB - We have conducted a multicenter collaborative retrospective analysis to evaluate clinical characteristics and to compare prognostic scoring systems of 149 Korean patients with myelodysplastic syndromes (MDS). The median age of the patients was 53 years (range 17-82 years) with high of the patients being younger than 40 years. Median survival was 22.6 months, and 25 patients (17%) progressed to acute myelogenous leukemia (AML) with a median interval of 6 months (range 1-45 months). Major independent variables assessed by multivariate analysis were FAB subtypes and bone marrow (BM) blast percentages for survival and BM blast percentages for AML transformation. To compare the various scoring systems in the prediction for survival and transformation to AML, FAB, Sanz and Bournemouth scoring systems were applied to all patients, while the international prognostic scoring system (IPSS), Lille and Toyama scoring systems were applied to 91 patients. The Sanz scoring system (P < 0.0001), FAB classification (P < 0.0001), IPSS (P < 0.001), and Toyama scoring system (P < 0.005) were highly predictive for survival showed greater discrimination than that of the other systems. For AML transformation, the IPSS (P < 0.0001), Toyama scoring system (P < 0.0001), FAB classification (P < 0.0001), and Lille scoring system (P < 0.005) successfully discriminated risk groups. Although the prognostic factors and the distribution of age were different from those in Western reports, the IPSS and Toyama scoring system were applicable for predicting survival and leukemic transformation in Korean patients with MDS. PMID- 10374858 TI - CD10 and CD19 fluorescence intensity of B-cell precursors in normal and leukemic bone marrow. Clinical characterization of CD10(+strong) and CD10(+weak) common acute lymphoblastic leukemia. AB - In order to assess the age-related changes in CD10 and CD19 fluorescence intensity (FI) the present study analyzed by flow cytometry 56 sternal biopsies from 'normal' infants, children and adults undergoing cardiac surgery. The CD10(+weak) subset was predominant in all age groups, representing approximately 50% of the bone marrow (BM) lymphoid cells in children younger than 4 years. Both CD10+ subsets significantly decreased with age but their ratio did not differ significantly. Moreover, the intensity of CD10 and CD19 fluorescence in the strong and weak subsets did not vary with age. The CD19 intensity was significantly higher in CD10(+weak) than in CD10(+strong) cells. In addition, we classified as CD10(+weak) or CD10(+strong) the leukemic cells from BM aspirates of 117 patients with common acute lymphoblastic leukemia (cALL) (78 children and 39 adults). A higher frequency of cases expressing the CD19+ CD10(+strong) phenotype was observed both in children and adults. Children of the CD10(+weak) group tended to be older than those of the CD10(+strong) group (median = 7 vs. 4 years, P = 0.07), and presented a significantly higher frequency of splenomegaly (93.7 vs. 55%, P = 0.04), which was massive in about 60% of these cases. Among adults, a significantly higher frequency of cases expressing the CD10(+weak) phenotype was observed in females. No other clinical or biological difference was detected between the two groups either for children or adults. Concerning the treatment outcome, we did not observe significant differences in complete remission rate (CRR) or in disease free survival (DFS) among the 32 children and 28 adults analyzed. Finally, we compared the CD10 and CD19 intensity in normal and leukemic BM. Overexpression of either or both antigens in leukemic cells was observed in 42.4% of the cALL cases. In these cases, using cut off values of 110 afu for the CD10 FI and of 100 afu for the CD19 FI, the detection of leukemic cells was possible at levels of 0.2% based on CD10 analysis, of 0.6% based on CD19, and 0.02% when both antigens were overexpressed. In conclusion, we demonstrated that the heterogeneity of CD10 and CD19 fluorescence intensity is of no clinical relevance in cALL, although its study may be helpful for the diagnosis and the detection of minimal residual disease. PMID- 10374859 TI - P-glycoprotein and terminal transferase expression identify prognostic subsets within cytogenetic risk classes in acute myeloid leukemia. AB - Clinical and biological features were assessed in 204 consecutive de novo adult acute myeloid leukemia (AML) patients who received intensive chemotherapy regimens. Multiparameter flow cytometric assays both of the multidrug resistance (MDR-1)-associated P-glycoprotein (PGP) using the UIC2 monoclonal antibody (MoAb), and of terminal transferase (TdT) were performed. Cytogenetic findings were obtained from 196 patients with high resolution banding. At onset, UIC2 and TdT positivities were detected in 58.5% and 24% of cases, respectively. There were strict correlations either between UIC2 negativity and FAB M3 or between TdT and FAB M0-M1 (P = 0.001 and < 0.0001, respectively). On the other hand, age was significantly associated with cytogenetic risk classes (P < 0.0001). CD34 positivity was highly correlated with TdT expression (P < 0.0001). Moreover, CD7 and CD11b were significantly represented in UIC2+ subset (P < 0.0001). Rhodamine 123 (Rh 123) efflux was significantly higher in 75 UIC2 positive patients compared to 65 UIC2 negative ones (P < 0.001). As regards to cytogenetics, TdT positivity was strongly related either to t(9;22) or single/associated anomalies of chromosome 7; on the other hand, most or all cases with t(8;21) or t(15;17) were UIC2 or TdT negative, respectively. The rate of first complete remission (CR) differed both between UIC2+ and UIC2- cases and between TdT+ and TdT- ones (40% versus 72%, P < 0.001; and 36% versus 61%, P = 0.001, respectively). The survival rates (Kaplan-Meier method) were significantly shorter either in UIC2+ or in TdT+ patients (P = 0.005 and = 0.011, respectively). UIC2 and TdT negative cases showed longer remission duration (P = 0.03 and = 0.22, respectively). The additional effect of UIC2 and TdT on prognosis allowed us to identify two subsets of patients, the first [UIC2- TdT-] at better and the second [UIC2+ TdT+] at worse clinical outcome compared to single UIC2 and TdT cases, concerning CR (P < 0.001), survival (P < 0.0001) and CR duration (P = 0.007). The combinations [UIC2+ TdT-] and [UIC2- TdT+] showed an intermediate clinical course. A strong difference was found between poor risk and intermediate/favorable risk cytogenetic classes with regard to CR rate (P < 0.0001), overall survival and CR duration (P < 0.001). Nevertheless, within the poor risk class, UIC2 positivity was able to identify patients at worst prognosis with regard to CR (P = 0.005), survival (P = 0.02) and CR duration (P = 0.015). On the other hand, UIC2 and TdT negativity allowed us to distinguish patients with longer survival (P = 0.012 and = 0.04, respectively) and CR duration (P = 0.04 and = 0.025, respectively) within the intermediate/favorable risk class. The independent prognostic value of UIC2, TdT and cytogenetic risk classes was confirmed in multivariate analysis. These results suggest that PGP and TdT expressions, together with cytogenetic findings, may represent a basic predictor of chemotherapeutic failure in AML. PMID- 10374860 TI - Functional analysis of P-glycoprotein and multidrug resistance associated protein related multidrug resistance in AML-blasts. AB - Despite the high effectiveness of various P-glycoprotein (P-gp) modulating substances in vitro their clinical value e.g. for combination treatment of acute myelogenous leukemias (AML) remains still unclear. This might be explainable by recent findings that other factors than P-gp (e.g. the multidrug resistance associated protein (MRP)) may also be involved in clinical occurring drug resistance. To study P-gp and MRP mediated MDR in AML blasts from patients with relapses at the functional level we measured rhodamine 123 (RHO) efflux in combination with a P-gp specific (SDZ PSC 833) or a MRP specific (MK571) modulator, respectively. Furthermore, direct antineoplastic drug action was monitored by determination of damaged cell fraction of a blast population using flow cytometry. We generally found strongly modulated RHO efflux by SDZ PSC 833 but slight RHO-efflux modulation by MK571 in blasts from relapsed states of AML expressing MDR1 or MRP mRNA at various levels. We could not demonstrate, though, significant PSC 833 or MK571 mediated modulation of the cytotoxic effects of etoposide. The results point to the possibility that combination of etoposide and a modulator might not improve responses to chemotherapy by targeting P-gp or MRP exclusively. PMID- 10374861 TI - ALL- and CML-type BCR/ABL mRNA transcripts in chronic myelogenous leukemia and related disorders. AB - Using the reverse transcription polymerase chain reaction, we investigated acute lymphoid leukemia (ALL)-type, and chronic myelogenous leukemia (CML)-type BCR/ABL mRNA expression in a total of 66 patients with chronic myeloproliferative disorder (CMPD). Thirty-six of 37 patients with CML were positive for CML-type mRNA. Thirteen of the 25 CML had ALL-type mRNA expression. The patients with ET, PV, MF, and CMML did not have any detectable BCR/ABL expression. The most remarkable finding was that two patients, a Ph1-positive CML patient and a patient with a presumptive diagnosis of essential thrombocythemia (ET), showed only ALL-type chimeric mRNA expression. PMID- 10374862 TI - Expression of LFA-1 identifies different prognostic subgroups in patients with advanced follicle center lymphoma (FCL). AB - In a retrospective immunohistochemical study based on 27 patients with stage IV follicle center lymphoma (FCL) the expression of CD44standard (CD44s), LFA-1 (CD11a, CD18), VLA-4 (CD49d, CD29) and ICAM-1 (CD54) was analysed on lymphoma cells in bone marrow infiltrates. The results were correlated to clinical data and overall survival. Our data demonstrate that the expression of LFA-1 on lymphoma cells is predictive for the prognosis of patients with advanced FCL. In detail, patients exhibiting weak to moderate expression (+/++) of CD11 and CD18 showed a significantly shorter median survival (51 months and 33 months, respectively) than did those presenting with strong expression ( ) of the LFA-1 adhesion molecule (P = 0.04 and P = 0.0051, respectively). Furthermore, multivariate analysis identified CD18 as a new independent prognostic factor in patients with advanced FCL. Our findings emphasize the relevance of adhesion molecules for the pathology of FCL and give further support for their impact on clinical course and overall survival. PMID- 10374863 TI - Hepatocyte growth inhibitory factor derived from HTLV-I(+) T-cell line is identical to IL-6. AB - We previously reported that the culture supernatant of the human T-cell leukemia virus (HTLV-I) infected-T-cell line--ATL-2--included factor(s), which had an inhibitory effect on epidermal growth factor (EGF)-stimulated proliferation of primary cultured rat hepatocytes. After crude purification, we arbitrarily named it hepatocyte growth inhibitory factor (HGI). In this study, we further purified HGI and determined its amino acid sequence. For purification, we used 4-steps column chromatography and SDS-PAGE. The purified proteins consisted of two bands of 20 and 27 kDa in SDS-PAGE analysis. Protein extracted from each band had an inhibitory effect on rat hepatocyte growth. Amino acid analysis of the purified 20 kDa band revealed that the 34 amino acids were identical to those of IL-6. The inhibitory effect of the factor was neutralized by an anti IL-6 neutralizing antibody. Using Western blot analysis of HGI, an anti IL-6 antibody recognized both 20 and 27 kDa bands. Consequently HGI was determined to be identical to IL 6, which occurred in higher levels in the sera of adult T-cell leukemia (ATL) patients. PMID- 10374864 TI - Constitutive expression of the Wilms' tumor gene WT1 inhibits the differentiation of myeloid progenitor cells but promotes their proliferation in response to granulocyte-colony stimulating factor (G-CSF). AB - Bone marrow (BM) cells that were concentrated for hematopoietic progenitor cells by in vivo treatment with 5-FU were infected with a recombinant retrovirus containing a human full-sized, non-spliced type WT1 (Wilms' tumor gene 1) cDNA and then colony-assayed in the presence of granulocyte-colony stimulating factor (G-CSF). Significantly more colony-forming units granulocyte-monocyte (CFU-GM), colony-forming units granulocyte (CFU-G), and colony-forming units monocyte (CFU M) colonies were formed in response to G-CSF from the BM cells infected with the WT1-containing retrovirus than from the control BM cells infected with an empty vector. Furthermore, FACS analysis of cell surface differentiation markers showed the inhibition of differentiation by constitutive WT1 expression resulting from the infection with the WT1-containing retrovirus. These results thus showed that the constitutive WT1 expression promoted the proliferation of myeloid progenitor cells but inhibited their differentiation in response to G-CSF, suggesting the alteration of G-CSF signaling pathway. The results also supported our hypothesis that the WT1 gene performs an oncogenic rather than a tumor suppressor gene function in hematopoietic progenitor cells, although the WT1 gene potentially performs both functions. This finding implies an important role of the WT1 gene in leukemogenesis. PMID- 10374865 TI - Augmentation of methylprednisolone-induced differentiation of myeloid leukemia cells by serine/threonine protein phosphatase inhibitors. AB - To elucidate the roles of serine/threonine protein phosphatases type 1 (PP1) and type 2A (PP2A) in methylprednisolone-induced differentiation of HL60 cells into granulocytes and K562 cells into monocytes, we examined the effect of serine/threonine protein phosphatase inhibitors, okadaic acid and Cal-A on the proliferation/differentiation of HL60 and K562 cells. Okadaic acid and Cal-A augmented methylprednisolone induced granulocytic differentiation and cell death of HL60 cells and monocytic differentiation and cell death of K562 cells in different dose ranges, respectively. These data suggest an important role of PP1 and PP2A in the mechanism leading to differentiation of leukemic cells. PMID- 10374866 TI - Unusual clinical presentation in a patient with myelodysplastic syndrome, with subsequent hematological remission and suppression of the malignant clone following treatment with cyclosporine A, erythropoietin and granulocyte colony stimulating factor. AB - A 35-year-old female presented with isolated thrombocytopenia of autoimmune origin. One and a half years later, hypoplastic myelodysplastic (MDS) was diagnosed. Following treatment with cyclosporin A, erythropoietin and granulocyte colony-stimulating factor, the patient has achieved a sustained hematological remission which is still ongoing after 3 years. Furthermore, to the best of our knowledge, this is the third case described in the literature where treatment with cytokines alone or in combination with immunosuppressive agents has resulted in a long standing cytogenetic response in MDS. PMID- 10374867 TI - Dendritic cell-based vaccine: a promising approach for cancer immunotherapy. AB - The unique ability of dendritic cells to pick up antigens and to activate naive and memory CD4+ and CD8+ T cells raised the possibility of using them to trigger a specific anti-tumor immunity. If numerous studies have shown a major interest in dendritic cell-based vaccines for cancer immunotherapy in animal models, only a few have been carried out in human cancers. In this review, we describe recent findings in the biology of dendritic cells that are important to generate anti tumor cytotoxic T cells in vitro and we also detail clinical studies reporting the obtention of specific immunity to human cancers in vivo using reinfusion of dendritic cells pulsed with tumor antigens. PMID- 10374868 TI - Primary effusion lymphoma after heart transplantation: a new entity associated with human herpesvirus-8. AB - Deep immunosuppression and Epstein-Barr virus (EBV) infection promote the emergence of lymphoproliferative disorders in patients undergoing solid organ transplantation. In the last few years a new herpesvirus, named human herpesvirus 8 (HHV-8), has been identified in Kaposi's sarcoma and primary effusion lymphoma (PEL) developing in AIDS patients. Subsequently, the same viral DNA sequences have been identified in almost all cases of Kaposi's sarcoma emerged outside HIV infection, thus suggesting their possible pathogenetic role in this tumor. Similarly, the association between HHV-8 and PEL also emerged in cases without HIV infection, even though the total number of these patients is still limited. Here, we focus on the emergence of this unusual lymphoma in patients undergoing solid organ transplant and underline once again its association with the HHV-8. Moreover, despite the characteristic local growth of this peculiar type of lymphoma, we demonstrate at the molecular level, an early neoplastic spread to the bone marrow suggesting the need to investigate in more detail the origin of the disease, as well as the molecular mechanisms controlling its systemic dissemination. PMID- 10374869 TI - Centric and pericentric chromosome rearrangements in hematopoietic malignancies. AB - Cytogenetic and fluorescence in situ hybridization (FISH) analysis of 10 patients with various hematopoietic malignancies revealed the presence of dicentric chromosomes or pericentric chromosome rearrangements. Dicentrics were only ascertained by FISH studies in six patients. Two types of pericentric chromosome rearrangements have been observed: 'classical' dicentrics with two clearly separated centromeric regions, and more unusual rearrangements with a breakpoint within the centromeric or heterochromatic area, but outside the alphoid domain. FISH analysis of partial chromosome 1 q duplications present in three Burkitt lymphoma cell lines confirmed the partial involvement of the non-alphoid centromeric domain in the duplicated chromosome segment. The incidence of centromeric and pericentromeric rearrangements in hematopoietic malignancies may be higher than hitherto admitted. The chromosomal localization of these rearrangements suggests several mechanisms possibly involved in the malignant process and deserves more systematic study. PMID- 10374870 TI - Cytogenetic studies of infant acute lymphoblastic leukemia: poor prognosis of infants with t(4;11) - a report of the Children's Cancer Group. AB - Infants less than 1 year of age at diagnosis of acute lymphoblastic leukemia (ALL) have a poor prognosis, which has been attributed primarily to a breakpoint in chromosomal band 11q23 or the MLL gene. Most infants with an 11q23 breakpoint have a t(4;11)(q21 ;q23). We studied the cytogenetics of the leukemia cells of 56 infants on CCG-1883, a single-arm clinical treatment protocol for infant ALL. Twenty-one patients had t(4;11)(q21;q23), seven had other rearrangements with breakpoints in 11q23 (other 11q23), 16 had normal chromosomes, two had t(1;19)(q32;p13), one had >50 chromosomes, and nine had non-recurring structural abnormalities. To determine whether there is a difference in outcome for infants with t(4;11), other 11q23 and the remaining patients, we compared event-free survival (EFS) and other clinical and laboratory features of the above infants. Infants without t(4;11) and those with other 11q23 rearrangements had significantly better EFS than those with t(4;11) (P= 0.007 and P= 0.02, respectively). t(4;11) correlated with age less than 6 months and with CD10 negativity, both of which also were poor prognostic indicators. After adjustment for age, there was still a significant difference in EFS between patients with t(4;11) and those with other 1lq23 rearrangements (P=0.02), and between patients with t(4;11) and those without t(4;11) (P=0.04). Among CD10 negative patients, t(4;11) was associated with a worse EFS (P=0.01). Multivariate analysis showed that after adjusting for a variety of clinical and laboratory features, t(4;11) was the most important prognostic factor for poor outcome, and patients with other 11q23 rearrangements had as good an outcome as the remaining patients without t(4;11). PMID- 10374871 TI - Expression of HOX genes, HOX cofactors, and MLL in phenotypically and functionally defined subpopulations of leukemic and normal human hematopoietic cells. AB - To explore the possibility that deregulated HOX gene expression might commonly occur during leukemic hematopoiesis, we compared the relative levels of expression of these and related genes in phenotypically and functionally defined subpopulations of AML blasts and normal hematopoietic cells. Initially, a semi quantitative RT-PCR technique was used to amplify total cDNA from total leukemic blast cell populations from 20 AML patients and light density cells from four normal bone marrows. Expression of HOX genes (A9, A10, B3 and B4), MEIS1 and MLL was easily detected in the majority of AML samples with the exception of two samples from patients with AML subtype M3 (which expressed only MLL). Low levels of HOXA9 and A10 but not B3 or B4 were seen in normal marrow while MLL was easily detected. PBX1a was difficult to detect in any AML sample but was seen in three of four normal marrows. Cells from nine AML patients and five normal bone marrows were FACS-sorted into CD34+CD38-, CD34+CD38+ and CD34-subpopulations, analyzed for their functional properties in long-term culture (LTC) and colony assays, and for gene expression using RT-PCR. 93 +/- 14% of AML LTC-initiating cells, 92 +/- 14% AML colony-forming cells, and >99% of normal LTC-IC and CFC were CD34+. The relative level of expression of the four HOX genes in amplified cDNA from CD34- as compared to CD34+CD38- normal cells was reduced >10-fold. However, in AML samples this down-regulation in HOX expression in CD34- as compared to CD34+CD38- cells was not seen (P < 0.05 for comparison between AML and normal). A similar difference between normal and AML subpopulations was seen when the relative levels of expression of MEIS1, and to a lesser extent MLL, were compared in CD34+ and CD34- cells (P < 0.05). In contrast, while some evidence of down-regulation of PBX1a was found in comparing CD34- to CD34+ normal cells it was difficult to detect expression of this gene in any subpopulation from most AML samples. Thus, the down-regulation of HOX, MEIS1 and to some extent MLL which occurs with normal hematopoietic differentiation is not seen in AML cells with similar functional and phenotypic properties. PMID- 10374872 TI - Expression of inducible nitric oxide synthase (NOS) in bone marrow cells of myelodysplastic syndromes. AB - Nitric oxide (NO) is a biological mediator which is synthesized from L-arginine by a family of nitric oxide synthases (NOS). We have studied the expression of the inducible NOS (iNOS) by bone marrow cells from the patients with myelodysplastic syndromes (MDS) at the mRNA level by RT-PCR assay and at the protein level by immunohistochemical staining using a specific anti-iNOS monoclonal antibody. The iNOS message was present in 92% of bone marrow tissues from MDS patients (11 out of 12) by an examination using RT-PCR. Basically, iNOS message was negative or very weak in control (1/9) and AML (0/7) cases. This was supported by immunohistochemical findings that the iNOS was positive in most of the bone marrow samples from MDS patients (9 out of 12), while bone marrow cells of control (O out of 12) and AML (O out of 5) cases were basically negative. Double immunostaining for CD68 antigen, which is a marker for macrophage lineage cells, and iNOS was performed on MDS bone marrow sections. iNOS was dominantly localized to bone marrow macrophages, although a part of myeloid cells were also positively stained with anti-iNOS antibody in a part of cases. These results indicated that there is some in vivo induction of iNOS expression for local NO production that might be involved in the dysregulation of hematopoiesis in bone marrow of MDS. PMID- 10374873 TI - Therapy-related adult acute lymphoblastic leukemia with t(4;11)(q21; q23): MLL rearrangement, p53 mutation and multilineage involvement. AB - A diagnosis of pro-B acute lymphoblastic leukemia (ALL) with CD15+ was made in a 42-year-old woman, 12 months after the treatment of uterine adenocarcinoma by carboplatinum, anthracyclines, etoposide and radiotherapy. Molecular cytogenetic studies revealed a karyotype with multiple chromosome changes, including the t(4;11)(q21;q23) and a 17p-chromosome, with MLL disruption and 17p13/p53 gene deletion in 86% of the cells. A p53 exon 6 mutation was documented, resulting in p53 protein stabilization, with 20% of the cells reacting with the 1801 anti-p53 monoclonal antibody. Dual-color FISH using MLL and p53 probes was performed on peripheral blood smears, providing direct evidence of the involvement of the blast cells and of the granulocytic lineage. Only a partial, shortlasting response was obtained by induction treatment, confirming that a poor prognosis is associated with therapy-related ALL with the 4;11 translocation. PMID- 10374874 TI - Discontinuous deletions at 11q23 in B cell chronic lymphocytic leukemia. AB - Loss of genomic material in 11q is one of the most common structural chromosome aberrations in B cell chronic lymphocytic leukemia (B-CLL). In order to characterize the deletions of 11q23 in B-CLL, we performed fluorescence in situ hybridization (FISH) with eight YAC (yeast artificial chromosome) probes on peripheral leukocytes of 30 patients. These YACs form a contig spanning 7.8 Mb at 11q23.1-q23.3. We found deletions in nine out of 30 cases (30%) and five of them had discontinuous deletions in this region. The region represented by YAC 755b11 (1.6 Mb in size) was involved in all cases with deletions, supporting the hypothesis that this region might contain a novel gene of pathogenic importance to B-CLL. A more distal region represented by YAC 785e12 (760 kb in size) was deleted frequently and specifically. Whether there is another novel gene of pathogenic importance to B-CLL and what is its potential relationship to the deletions in the region represented by YAC 755b11, are issues that require further studies. PMID- 10374875 TI - Detection of inducible nitric oxide synthase (iNOS) mRNA by RT-PCR in ATL patients and HTLV-I infected cell lines: clinical features and apoptosis by NOS inhibitor. AB - Various tumors have been reported to express an inducible form of nitric oxide synthase (iNOS), and nitric oxide (NO) may affect the clinicopathological features of these tumors. Previously, Burkitt's lymphoma and Epstein-Barr virus (EBV)-infected cells were shown to express iNOS constitutively at a low level. We analyzed iNOS expression by the reverse transcriptase-polymerase reaction method (RT-PCR) in eight HTLV-I-infected cell lines (five were ATL-derived lines and there were in vitro transformed lines), nine ATL patients (three were chronic, two were acute, and four were lymphoma type), and an HTLV-I-negative T cell line (CEM). In four ATL derived and in all three in vitro transformed cell lines, iNOS was expressed constitutively, but it was not expressed in CEM cells. Four out of nine ATL patients also showed iNOS expression. The expression of iNOS was found in all subtypes of ATL. Three of four iNOS-positive patients had infiltration of ATL cells to organs such as skin, lung, or liver. In NOS inhibitor (NG-monomethyl L-arginine: L-NMMA)-containing medium, an iNOS-positive ATL cell line (K3T) showed growth inhibition and DNA ladder. Although only a limited number of patients was analyzed, our results suggest that NO may be involved in the invasive character of ATL cells. The NOS inhibitor can induce apoptosis in an ATL cell line, as it does in EBV-infected cell lines. PMID- 10374876 TI - Cleavage of Bcl-2 is an early event in chemotherapy-induced apoptosis of human myeloid leukemia cells. AB - The proto-oncogene product Bcl-2 protects a wide variety of cell types from apoptosis via a hitherto unknown mechanism. Bcl-2 has been shown to function upstream of the death proteases (caspases) in some, but not all, occurrences of apoptotic cell death. Using the myeloid leukemic cell line P39 we report the chemotherapy-induced caspase-dependent cleavage of endogenous Bcl-2. Etoposide treatment of these cells triggered a time-dependent activation of type II and type III caspases and cleavage of Bcl-2 yielding a 23 kDa cleavage fragment. The emergence of this cleavage product was blocked by the general caspase inhibitor zVAD-fmk, as well as the type III caspase inhibitor IETD-fmk and the caspase-9 selective inhibitor LEHD-fmk, while the type II caspase inhibitor DEVD-fmk proved considerably less efficient. Bcl-2 cleavage preceded cleavage of the known caspase-3 substrate, poly(ADP-ribose) polymerase (PARP), as well as that of the caspase-6 substrate, lamin B, indicating that Bcl-2 cleavage is a relatively early event in the apoptosis cascade in this experimental model. While evidence for cleavage of Bcl-2 in several subcellular compartments of etoposide-treated cells was obtained, this cleavage was detected predominantly in the mitochondrial fraction, thus providing further support for the central role of mitochondria in apoptosis. Caspase-mediated cleavage following etoposide treatment of these myeloid leukemic cells may represent a means for the attenuation of Bcl-2 function upon apoptosis induction. PMID- 10374877 TI - Induction of apoptosis by bestatin (ubenimex) in human leukemic cell lines. AB - We investigated the growth inhibitory activity of bestatin, an inhibitor of aminopeptidase N (CD13), on six human leukemic cell lines. Proliferation of all the cell lines except KG1 was inhibited by bestatin. P39/TSU, HL60 and U937 were highly sensitive, with 50% growth inhibitory concentrations (IC50) close to the maximum serum concentration when bestatin was orally administered at 30 mg in clinical application. All cell lines except for K562 highly expressed CD13, but a clear correlation between the sensitivity to bestatin and expression of CD13 was not observed. Other aminopeptidase inhibitors such as amastatin A, arphamenine B and WM15 antibody showed no growth inhibitory effects. To confirm the growth inhibitory effects of bestatin, we quantitatively examined DNA fragmentation in five bestatin-sensitive cell lines. Bestatin dose-dependently induced DNA fragmentation in those cell lines. In case of U937, bestatin induced DNA fragmentation quantitatively and DNA ladder and enhanced caspase-3 activity. Furthermore, the growth inhibition by bestatin was reduced by the caspase inhibitor Z-Asp-CH2-DCB. These results suggested that bestatin exhibits direct antileukemic effects against human leukemic cell lines through the induction of apoptosis. PMID- 10374878 TI - Inhibition of constitutively activated Stat3 correlates with altered Bcl-2/Bax expression and induction of apoptosis in mycosis fungoides tumor cells. AB - The Jak/Stat signaling pathway transmits signals from many cytokine and growth factor receptors to target genes in the nucleus. Constitutive activation of Stat3 has recently been observed in many tumor cells and dysregulation of the Stat signaling pathway has been proposed to be implicated in malignant transformation. In a previous study, we found constitutively tyrosine phosphorylated Stat3 in mycosis fungoides tumor cells. Here, we show that the Jak kinase inhibitor, Ag490, inhibits the constitutive binding of Stat3 to an oligonucleotide representing the Stat-binding sequence from the ICAM promotor. The decreased ability of Stat3 to bind DNA precedes dynamic alterations in the expression of anti-apoptotic Bcl-2 and pro-apoptotic Bax proteins (decreased Bcl-2 expression and increased Bax expression) and induction of apoptosis. Thus, our data suggest that the involvement of Stat3 in oncogenic transformation could be mediated through regulation of survival signals. PMID- 10374880 TI - Chromatin structure and transcriptional regulation of the stem cell leukaemia (SCL) gene in mast cells. AB - The stem cell leukaemia (SCL) gene is a member of the basic helix-loop-helix family of transcription factors and is essential for the development of all haematopoietic lineages. SCL is expressed in pluripotent haematopoietic stem cells and also following commitment to the erythroid, mast and megakaryocytic lineages. The mechanisms responsible for this pattern of expression are poorly understood, but are likely to illuminate the molecular basis for stem cell development and lineage commitment. Here we present the first description of the regulation of the SCL gene in mast cells. In this study we systematically analysed the chromatin structure of a 45 kb region of the murine SCL locus in mast cells. The pattern of DNase 1 and restriction endonuclease hypersensitive sites in mast cells was distinct from, but overlapped with, the pattern previously described in erythroid and primitive myeloid cells. Each potential regulatory element was tested using transient reporter assays to assess their functional significance in mast cells. These studies identified two potent enhancers, one of which was downstream of the SCL gene. Further characterisation of this 3' enhancer demonstrated that it required the presence of two distinct DNase 1 hypersensitive sites for full activity, and that it was capable of stimulating transcription from both promoter 1a and 1b. Since the 3' enhancer is active in both erythroid and mast cells, it will now be important to see whether it is independently activated in these lineages, or whether it is also active in haematopoietic stem cells. PMID- 10374879 TI - In acute promyelocytic leukemia NB4 cells, the synthetic retinoid CD437 induces contemporaneously apoptosis, a caspase-3-mediated degradation of PML/RARalpha protein and the PML retargeting on PML-nuclear bodies. AB - CD437-induced apoptosis has been investigated in NB4, a human t(15;17) acute promyelocytic leukemia (APL) cell line, and in the retinoic acid (RA)-resistant NB4-R1 derivative subclone. Both NB4 and NB4-R1 cells underwent rapid apoptosis in response to low doses of CD437 (10(-7)M). This apoptosis did not require the activation of classical retinoid receptors and like arsenic (As)-induced apoptosis was preceded by the rapid activation of a caspase-3-like enzymatic activity as indicated by the increase of DEVD-pNA hydrolytic activity, by the processing of procaspase-3 protein and by the cleavage of poly(ADP-ribose) polymerase (PARP). Furthermore, it was demonstrated that the caspase-3-like proteolytic activity is responsible for the degradation of both the PML/RARalpha oncogenic protein and the normal RARalpha proteins. In CD437-treated cells, PML proteins were not degraded and PML relocalization on PMLNBs occurred in all the cells before death. CD437-induced apoptosis and receptor degradation were proteasome independent and not influenced either by inhibitors of protein tyrosine kinases (PTK), protein tyrosine phosphatases (PTPases) and serine proteases or by glutathione levels. Moreover, our data suggested that as for As2O3-induced apoptosis Bc12 modulation is not significant for CD437-induced apoptosis of NB4 cells. Since CD437 induces in vitro the rapid apoptosis of both RA-sensitive and -resistant APL cells, it could represent the first retinoid potentially able to eradicate in vivo malignant leukemia blasts. PMID- 10374881 TI - APS, an adaptor protein containing Pleckstrin homology (PH) and Src homology-2 (SH2) domains inhibits the JAK-STAT pathway in collaboration with c-Cbl. AB - We cloned a novel adaptor protein, APS (adaptor molecule containing Pleckstrin homology (PH) and Src Homology-2 (SH2) domains), which was tyrosine phosphorylated in response to c-kit or B cell receptor stimulation. Here, we report that APS was tyrosine phosphorylated by Janus kinase-2 (JAK2) at its C terminal tyrosine residue and interacted with c-Cbl. Forced expression of APS in an erythropoietin (EPO)-dependent hematopoietic cell line resulted in reduced activation of STAT5 but not cell proliferation in response to EPO. APS bound to the phosphorylated tyrosine residue, Y343 of the erythropoietin receptor cytoplasmic domain. Co-expression of APS and c-Cbl, but not expression of either alone inhibited EPO-dependent STAT5 activation in 293 cells. This required the C terminal phosphorylation site, as well as PH and SH2 domains of APS. Therefore, one of the major functions of APS is in recruitment of c-Cbl into the receptor/JAK complex, thereby inhibiting JAK signaling activity. PMID- 10374882 TI - Growth inhibition, cytokinesis failure and apoptosis of multidrug-resistant leukemia cells after treatment with P-glycoprotein inhibitory agents. AB - The multidrug transporter P-glycoprotein (Pgp), which is frequently overexpressed in multidrug resistant leukemia, has many proposed physiological functions including involvement in transmembraneous transport of certain growth-regulating cytokines. Therefore, we studied cell growth of three pairs of drug resistant and sensitive leukemia cell lines (KG1a, K562 and HL60) exposed to three different inhibitors of Pgp. The resistant KG1a and K562 sublines, which expressed high levels of Pgp, responded to low doses of the cyclosporin SDZ PSC 833, the cyclopeptolide SDZ 280-446, and the cyclopropyldibenzosuberane LY335979 with a dose-dependent growth inhibition. In the resistant variants of KG1a and K562 cells the mean half-maximal growth inhibitory doses (GI50) of SDZ PSC 833 were 312 (SE 41) and 414 (SE 50) nM, those of SDZ 280-446 were 685 (SE 51) and 578 (SE 54) nM, and those of LY335979 were 66 (SE 1) and 48 (SE 8) nM, respectively. Exposure to 1 microM SDZ PSC 833 resulted in tetraploidization, cytokinesis failure and apoptosis of the KG1a and K562 MDR variants. Conversely, parental cells with no or low levels of Pgp and the non-Pgp resistant variant of HL60 cells were not receptive to these cytotoxic effects. We conclude that inhibition of Pgp may exercise selective cytotoxicity in Pgp-rich leukemia cells indicating a possible therapeutic target in multiresistant leukemia. PMID- 10374883 TI - The KOR-SA3544 antigen predominantly expressed on the surface of Philadelphia chromosome-positive acute lymphoblastic leukemia cells is nonspecific cross reacting antigen-50/90 (CD66c) and invariably expressed in cytoplasm of human leukemia cells. AB - We previously reported a novel monoclonal antibody KOR-SA3544 which predominantly reacted with a surface antigen (sSA3544) expressed on Philadelphia chromosome (Ph1)-positive acute lymphoblastic leukemia (ALL). In the present study, we demonstrate that the antibody specifically recognized nonspecific cross-reacting antigen (NCA)-50/90 (CD66c), one of the carcinoembryonic antigen (CEA)-related glycoproteins encoded by a member of the CEA gene family. In addition, we show that the SA3544 antigen (NCA-50/90) was invariably expressed in cytoplasm of all of the human leukemic cell lines examined (sSA3544-positive B-lymphoid two, sSA3544-negative T or B-lymphoid and non-lymphoid 24) regardless of the presence or absence of surface expression of this antigen. Immunoelectromicroscopic examination revealed that the cytoplasmic antigen was mainly present in granules in sSA3544-positive leukemia cells, whereas it was diffusely present in cytosol in sSA3544-negative leukemia cells. Thus, among members of the CEA family, NCA 50/90 was first demonstrated to be expressed not only on the surface of some leukemia cells, but also in cytoplasm of various types of leukemia cells. PMID- 10374884 TI - Early allogeneic transplantation for refractory or relapsed acute leukaemia following remission induction with FLAG. AB - The prognosis for patients with secondary AML, primary resistant AML or ALL and early (<12 months) relapse of acute leukaemia remains extremely poor with conventional chemotherapy. As part of a strategy to improve the outcome for these patients we have treated 22 consecutive patients (18 AML, four ALL, median age 35 years) with either primary resistant disease (n=3), early relapsed leukaemia (n= 12) or secondary AML (n= 7, four RAEBt, two antecedant ALL and one antecedant Hodgkin's disease) with 'FLAG' induction chemotherapy with the aim of proceeding to early allogeneic transplantation either from sibling or unrelated donors. Eighteen patients achieved CR after one course of FLAG, including five patients who had documented p-glycoprotein-induced multidrug resistance and 10 patients with adverse cytogenetic abnormalities. Eight patients were consolidated with a second course of FLAG prior to transplantation and so far 16 patients have undergone allogeneic transplantation, 10 from unrelated donors and six from sibling donors (one mismatched). By the time of transplant three patients had progressed and were in early relapse and all have relapsed post BMT. Of the remaining 13 patients transplanted in remission, nine remain in CCR at a range of 4-26 months, three have died of transplant-related complications (18%) and one patient has relapsed. We conclude that the use of FLAG induction therapy followed by early allogeneic transplantation from either a sibling or unrelated donor can be an effective strategy for the treatment of this difficult group of young patients with poor risk acute leukaemia and appears to be associated with a low procedure-related risk. PMID- 10374885 TI - Frequent chromosome arm 13q deletion in aggressive non-Hodgkin's lymphoma. AB - To clarify the role of allelic loss on chromosome arm 13q in lymphomagenesis, we performed fluorescence in situ hybridization (FISH) analysis of a total of 43 primary lymphomas, including both indolent and aggressive non-Hodgkin's lymphoma (NHL) and Hodgkin's disease (HD), using the specific probes at RB1 and D13S319 loci on the centromeric portion of chromosome arm 13q. Monosomy at either or both RB1 and D13S319 loci was detected in 15 of 43 (35%) lymphomas (14 of 43 cases at RB1 locus and seven of 43 cases at D13S319 locus); the 13q deletion was frequently detected in the aggressive NHLs (40%; 12 of 30 cases) compared with that in indolent NHL (17%; one of six cases) and a subset of HD (29%; two of seven cases). There are only six cases of 43 which have total monosomy 13q14, all aggressive NHL, 14% of total or 20% of this subgroup. In addition, we analyzed the loss of heterozygosity in 15 of the 43 primary lymphoma samples for several polymorphic microsatellite loci (D13S168, RB1 and D13S272) on the chromosome arm 13q, and confirmed the 13q deletion in four of five cases that were positive on FISH analysis. The subchromosomal region frequently altered in lymphoma on 13q14 is the region around RB1 locus and centromeric to D13S319 locus, which is an overlapped region frequently deleted in chronic lymphocytic leukemia. Together, our data indicate that the 13q alterations are present in a variety of types of lymphoma and occur in a significant proportion of aggressive NHLs, suggesting the possible presence of common candidate gene(s) on the 13q14 region, whose alteration may play an important role in the formation or development of a wide variety of mature lymphoid malignancies. PMID- 10374886 TI - Gastric low-grade MALT lymphoma, high-grade MALT lymphoma and diffuse large B cell lymphoma show different frequencies of trisomy. AB - Gastric MALT lymphoma is a distinct entity related to Helicobacter pylori gastritis. Some studies suggest a role for trisomy 3 in the genesis of these lymphomas, but they mainly focused on low-grade MALT lymphoma. Gastric MALT lymphoma, however, comprises a spectrum from low- to high-grade cases. Furthermore, its exact relation to primary diffuse large B cell lymphoma (DLBCL) of the stomach is not clear. We applied in situ hybridisation (ISH) with centromeric probes on 43 samples of 39 patients with primary gastric lymphoma (13 samples with low-grade MALT lymphoma, 25 with high-grade MALT lymphoma and five with DLBCL) to detect numerical aberrations of 10 chromosomes. ISH was performed immunohistochemically on nuclei isolated from paraffin-embedded resection tissue and on whole paraffin sections using immunofluorescence. In six of 13 low-grade MALT lymphomas trisomy was detected (46%) and mostly involved chromosome 3 (33%). In high-grade MALT lymphomas, trisomies were found in 16 of 25 cases (64%), mainly involving chromosomes 12 and 18. Trisomy 3 was present in only 13% of these cases. Of five DLBCL, only one showed trisomy. Nine of the 16 aberrant high grade MALT lymphomas (56%) showed trisomy of more than one chromosome per case vs two of six for low-grade cases. In lymphomas with separate low- and high-grade tumour components some trisomies were detected in both components, whereas others occurred only in the high-grade tumour cells. This supports the hypothesis that high-grade MALT lymphomas can develop from a low-grade type and that this progression is accompanied by the acquisition of more genetic aberrations. However, trisomy 3 probably does not play a major role in this progression. PMID- 10374887 TI - Analysis of the INK4a/ARF locus in non-Hodgkin's lymphomas using two new internal microsatellite markers. AB - Inactivation of the INK4a/ARF locus is a frequent event in non-Hodgkin's lymphomas (NHLs), which may be attributed to deletion, point mutation, and 5' CpG methylation at its promoter region. In the present study we evaluated the occurrence of deletions and genetic instability of INK4a/ARF locus in 30 paired normal and tumor samples of B cell NHLs by conducting an allelotypic analysis with two new polymorphic markers, one located at the intron 1 of p16INK4a gene and the other one placed downstream exon 1beta of p19ARF. Comparison of these results with those obtained in a previous paper using flanking markers (D9S171, D9S942, D9S958 and IFNA) allowed us to detect two new cases of microsatellite instability (L-446 and L-442), and to confirm the occurrence of LOH at the INK4a/ARF locus in one tumor (M-3770). On the contrary, this locus is not affected in three different tumors (L-421, L-272 and L-159) which exhibited LOH at some of the flanking markers. PMID- 10374888 TI - Profiles and prognostic values of LDH isoenzymes in patients with non-Hodgkin's lymphoma. AB - Serum lactic dehydrogenase (LDH) is an important prognostic factor in patients with non-Hodgkin's lymphoma (NHL). We have examined the LDH isoenzyme content in serum and CSF of patients with NHL, at diagnosis and at relapse. In patients with increased serum LDH at diagnosis, the percentage of isoenzyme 2 was increased in 52% of patients and the absolute value of isoenzyme 3 was increased in 64% of patients. In relapsing patients these values were respectively 69% and 65%. Conversely in patients with increased serum LDH due to myeloid regeneration after chemotherapy, isoenzymes 4 and 5, but not isoenzymes 2 or 3, were increased. High absolute values of isoenzyme 3 were correlated with an altered performance status, advanced tumor stage, and aggressive histology whereas high isoenzyme 2 percentages were correlated with altered performance status only. Among patients with high total serum LDH, a high content of isoenzyme 2 and a high absolute value of isoenzyme 3 were correlated with high serum levels of TNFalpha and TNF receptor p75. Analysis of total LDH and LDH isoenzyme profiles in CSF did not reveal any correlation with meningeal involvement by lymphoma. High isoenzyme 2 percentages and high absolute values of isoenzyme 3 in serum were both significantly associated with a shorter freedom-from-progression and overall survival. Isoenzyme 3 remained a prognostic factor for survival even when considering only patients with high total serum LDH at diagnosis. We conclude that there are some characteristic serum LDH isoenzyme profiles in patients with NHL and that some of these specific alterations may help refine the prognostic value of total serum LDH. PMID- 10374890 TI - Bone marrow transplant and toxoplasmosis. PMID- 10374889 TI - Detection of BCR-ABL transcripts in chronic myeloid leukemia (CML) using an in situ RT-PCR assay. AB - Methods of minimal residual disease (MRD) detection in chronic myelogenous leukemia (CML) include chromosome analysis, reverse transcriptase polymerase chain reaction (RT-PCR) and fluorescence in situ hybridization (FISH) techniques. We report a novel method to detect intracellular BCR-ABL messenger on single cells using in situ RT-PCR, which can be performed on blood and marrow slides, without extraction of the nucleic acid. After cellular permeabilization and fixation, the mRNA BCR-ABL was reverse transcribed and amplified by PCR using digoxigenin-labelled dUTP. The reaction was revealed with the anti-digoxigenin FITC antibody. On the fluorescent microscope, a strong positive green fluorescence signal was observed in 98-99% cells in Ph1-positive cell lines. A faint signal was detected in 1.5% and 2% of negative cell lines. Likewise, a faint signal was found in 1.6-2.8% of the cells in five normal controls (mean 2.2 +/- 1.1%). The positive threshold for in situ RT-PCR was therefore determined as mean + 2 s.d. = 4.4% cells. We used in situ RT-PCR by comparison to cytogenetics (at least 30 mitoses examined), and two-step RT-PCR (10(-6) sensitivity in our hands) in bone marrow samples from 13 CML patients: two patients at diagnosis and 11 patients in hematological remission after alpha interferon (three patients), hydroxyurea (one patient) autologous bone marrow transplantation (BMT) (one patient) and allogeneic BMT (six patients). In the two diagnostic patients, 90 and 95% cells were respectively strongly positive by in situ RT-PCR. In the six patients treated by allogeneic BMT, the median percentage of positive cells was 2.4% (range 1.8-3.2). All six patients had normal karyotype and negative two-step RT-PCR results. In the five other patients, two were treated by hydroxyurea alone or autologous BMT, and 11 and 13% of the cells were strongly positive; three were treated with interferon and 14-62% of the cells were positive, generally weakly. All five patients had persistence of Ph1 (in 9-56% mitoses), and positive RT-PCR results after one round. In conclusion, in situ RT-PCR can specifically identify cells with BCR-ABL transcript and its results are concordant with those of karyotype and RT-PCR. Because of its limited sensitivity and specificity, however, it appears to have limited value in the analysis of MRD. On the other hand, it can evaluate the presence and intensity of BCR-ABL fusion transcript at the single cell level, and this could be useful in treatment monitoring. PMID- 10374891 TI - A case of TCRgammadelta+ T-ALL with cross-lineage CD19 expression. PMID- 10374892 TI - Hodgkin's disease type Richter's syndrome in chronic lymphocytic leukemia. PMID- 10374893 TI - Immunoglobulin light chain variable region genes in multiple myeloma. PMID- 10374894 TI - ProMMP-9 (92 kDa gelatinase) in vitreous fluid of patients with proliferative diabetic retinopathy. AB - Matrix metalloproteinases (MMPs) are implicated in tissue destruction during various pathophysiologic conditions. The vitreous body is a gel-like extracellular matrix that undergoes liquefaction during aging and pathological processes. To investigate the pathogenic role of MMPs in proliferative diabetic retinopathy (PDR), we studied 73 eyes from PDR patients and 25 eyes from patients with non-diabetic ocular diseases. Vitreous MMPs were measured by zymography. Retinopathy was assessed by ophthalmoscopy and PDR was classified into 3 stages, 'naked', 'active', and 'quiescent'. Although proMMP-9 was expressed in only 8% (2/25) of non-diabetic patients, it was expressed in more than 80% (38/47) of 'active' PDR patients and still expressed in 60% (9/15) of those with 'quiescent' PDR. Vascular endothelial growth factor (VEGF) in vitreous fluids was undetectable (<0.16 ng/ml) in most of the non-diabetic patients, and was maximally elevated in the 'active' PDR patients (mean=2.20 ng/ml, range; 0.16 7.61), declining in patients with 'quiescent' PDR (1.04 ng/ml, 0.16-3.77). These results suggest that MMP-9 is one of the noteworthy factors in relation to the progress of PDR, as well as angiogenic cytokines such as VEGF. PMID- 10374895 TI - Effect of vitamin E and allylamine on the proliferation of cultured aortic smooth muscle cells from streptozotocin-induced diabetic rats. AB - To investigate the effect of vitamin E on the proliferation activity of vascular smooth muscle cells (SMCs) in diabetes mellitus, [3H]-thymidine incorporation was measured in cultured SMCs isolated from normal and streptozotocin-induced diabetic rats treated with or without vitamin E and/or allylamine. Untreated diabetic rats demonstrated significantly elevated concentrations of serum total cholesterol, triglycerides and malondialdehyde (MDA). Allylamine caused a further increase in serum MDA. Treatment with vitamin E decreased the serum concentrations of triglycerides and MDA in both allylamine-treated and -untreated diabetic rats. [3H]-Thymidine incorporation in cultured SMCs from diabetic rats was significantly increased compared with that from normal rats. SMCs from allylamine-treated diabetic rats showed an enhanced increase in thymidine incorporation compared with that from untreated diabetic rats. The increase in thymidine incorporation in SMCs from untreated and allylamine-treated diabetic rats was significantly reduced by the treatment with vitamin E. These observations suggest that vitamin E has a preventive effect on the proliferation of vascular SMCs in diabetes, and that this effect may be mediated through an enhancement of free radical scavenging. PMID- 10374896 TI - Differential modulation of alpha2-adrenoceptor subtypes in rat kidney by chronic desipramine treatment. AB - The profile of [3H]RX821002 (2-methoxy idazoxan) binding to alpha2-adrenoceptor subtypes in rat kidney membranes was evaluated in controls and after chronic treatment with desipramine (10 mg/kg, i.p., every 12 h, 7 days) or clorgyline (2 mg/kg, i.p., every 24 h, 21 days). [3H]RX821002 recognized with high affinity (Kd=1.5+/-0.2 nM in controls) a single and saturable population of binding sites (Bmax=57+/-5 fmol/mg protein in controls). The competitions by (-)-adrenaline, the alpha2B-adrenoceptor selective drug ARC239 (2-[2-[4-(o-methoxyphenyl) piperazin-1-yl]-ethyl]-4,4-dimethyl-1,3 (2H,4H)-isoquinolindione) and the alpha2A adrenoceptor selective drug BRL44408 (2-[2H-(1-methyl-1,3 dihydroisoindole)methyl]-4,5-dihydroimidaz ole) suggested the existence of both alpha2A- and alpha2B-adrenoceptors together with a non-adrenoceptor binding site. After chronic desipramine but not after chronic clorgyline treatments, the density (Bmax) of alpha2-adrenoceptors was increased (46%). In the presence of ARC239 (50 nM), the density of alpha2A-adrenoceptors increased (44%) in the desipramine-treated group without changes in the clorgyline-treated group. Conversely, in the presence of BRL44408 (100 nM), the density of alpha2B adrenoceptors was not affected by the treatments. The selective upregulation of the alpha2A-adrenoceptor subtype following chronic desipramine administration is compatible with a differential location and function of the alpha2-adrenoceptor subtypes in the rat kidney. PMID- 10374897 TI - Dual effect of female sex steroids on drug-induced gastroduodenal ulcers in the rat. AB - Since the sexual dimorphism of gastroduodenal ulcers is well known and might possibly relate to the actions of sex hormones, we studied the role of the female sex steroids, progesterone and 17beta-estradiol in cysteamine-induced mucosal ulcers in female Wistar rats (200-220 g). Administration of cysteamine (400 mg/kg, s.c.) provoked macroscopic gastroduodenal mucosa injury as assessed planimetrically, an increase in microvascular permeability in the stomach and the duodenum as assessed by extravasation of radiolabelled albumin, and decreased gastroduodenal mucus levels as assessed by the Alcian blue technique. Ovariectomy (2 weeks before cysteamine) decreased plasma 17beta-estradiol level as assessed by radioimmunoassay, gastroduodenal macroscopic injury and albumin extravasation, and increased mucus levels following cysteamine challenge. Administration of progesterone (10-50 mg/kg/week, s.c.) attenuated in a dose-dependent manner cysteamine-induced gastroduodenal mucosa injury and microvascular leakage, while it increased mucus levels in the stomach and the duodenum. In contrast, administration of 17beta-estradiol (1-5 mg/kg/week, s.c.) dose-dependently augmented gastric and duodenal macroscopic mucosa lesions and microvascular injury provoked by cysteamine, and caused a further reduction in gastroduodenal mucus levels observed after cysteamine administration. In different experiments, ovariectomy decreased indomethacin-induced gastroduodenal injury. The injection of 17beta-estradiol (1-5 mg/kg/week) did not affect gastroduodenal damage, while treatment with progesterone (10-50 mg/kg/week) protected against indomethacin provoked mucosa ulcers. It is concluded that female sex steroids play a role in drug-induced gastroduodenal ulcers by modulating microvascular permeability and mucus secretion. PMID- 10374898 TI - Affinity profiles of various muscarinic antagonists for cloned human muscarinic acetylcholine receptor (mAChR) subtypes and mAChRs in rat heart and submandibular gland. AB - A family of five subtypes of muscarinic acetylcholine receptors (mAChR) has been identified based on their molecular structures and second signal transduction pathways. In the present study, we examined the antagonist binding profiles of 9 muscarinic antagonists (atropine, 4-DAMP, pirenzepine, oxybutynin, tiquizium, timepidium, propiverine, darifenacin and zamifenacin) for human muscarinic acetylcholine receptor subtypes (m1, m2, m3, m4 and m5) produced by using a baculovirus infection system in Sf9 insect cells, and rat tissue membrane preparations (heart and submandibular gland). In a scopolamine methyl chloride [N methyl-3H]- ([3H]NMS) binding assay, pirenzepine and timepidium displayed the highest affinities for the m1 and m2 subtypes, respectively, and both zamifenacin and darifenacin had the highest affinities for the m3 subtype, although the selectivities among the five subtypes were less than 10-fold. Propiverine showed a slightly higher affinity for the m5 subtype, whereas none of the drugs used in this study was uniquely selective for the m4 subtype. The binding affinities of muscarinic antagonists for rat heart and submandibular gland strong correlated with those for human cloned m2 and m3 subtypes, respectively. These data suggest that [3H]NMS binding studies using rat heart and submandibular gland might be useful methods which predict the affinities of test drugs for human muscarinic M2 and M3 receptor subtypes. PMID- 10374899 TI - Plasma homocysteine levels increase in women during psychological stress. AB - Homocysteine is an amino acid that has been strongly associated with vascular disease. Plasma homocysteine concentrations are known to vary with dietary patterns and to decrease with exogenous estrogen use, but no other behavioral factors have been examined as potential modifiers of this risk factor. Because psychological stress has also been implicated in the development of cardiovascular disease, the purpose of this study was to test the hypothesis that acute psychological stress induces elevations in plasma homocysteine concentrations. A secondary aim was to test potential differences in response between premenopausal and postmenopausal women. Thirty-four healthy women, one half of whom were naturally postmenopausal with no hormone replacement, participated in this study. The psychological stressors included standard mental arithmetic and speech stressors. Blood samples were taken prior to, during, and after the stressors, and heart rate and blood pressure were also monitored. Results indicated significant elevations in plasma homocysteine during acute psychological stress, with a return to baseline concentrations during recovery. The pattern of findings for blood pressure and heart rate was similar, suggesting that the rise in homocysteine concentrations may have been sympathetically mediated. No effects of menopausal status or endogenous estrogens were found. The findings provide preliminary evidence that plasma homocysteine may be an important factor in the relationship between psychological stress and risk for heart disease. PMID- 10374901 TI - Catecholamine concentration in rat liver after high level transection of the spinal cord. AB - The high level transection of the spinal cord (C-7) provokes a sustained increase of rat liver catecholamines: biphasic increase in norepinephrine level 1 hour and 24 hour after the operation and 7-fold increase of dopamine content 4 hour after the chordotomy. In contrast to cervical transection, sham operation causes only an initial catecholamine increase, the maximum being at the first hour after the surgery. Our experimental data indicate a possible participation of cervical spinal pathways in regulation of liver catecholamine content. It is also shown that bilateral adrenalectomy augments liver norepinephrine concentration in spinal rats as compared to the non-adrenalectomized ones. The results presented here indicate that cervical chordotomy affects the functioning of the sympatho adrenal system, thus provoking specific changes in liver catecholamine content. The potential effect of such changes on a liver metabolic system (tyrosine aminotransferase induction) is discussed. PMID- 10374900 TI - Protein kinase C isoforms during the development of deciduomata in pregnant rats. AB - In this study, we determined the expression of protein kinase C (PKC) isoforms during pregnancy. At pregnant duration, PKC alpha was down-modulated in the deciduomata but not in the myometrium. Down-modulation was compatible with the increase in cell mitosis, which reached a maximum at 8-9 days. On the other hand, PKC zeta was not down-modulated. It was increased both in the cytosolic and particulate fractions of the deciduomata, and paralleled the frequency of decidual cell mitosis. The other PKC isoform of delta was also increased, but it was associated with the cell regression. Therefore, these findings confirmed that the variable expression of PKC isoforms in decidualizing tissue may be involved in the modulation of decidual cell growth. PMID- 10374902 TI - Diazepam reduces action potential duration in guinea-pig papillary muscle by a cAMP-dependent mechanism. AB - In this study, we have analyzed the role of cyclic AMP (cAMP) as the mediator of the decrease in action potential duration induced by diazepam. Diazepam (1-100 microM) reduced, in a dose-dependent manner, the duration of intracellular action potential recorded in the papillary muscle obtained from the right ventricle of the guinea pig heart. This effect was mimicked by the analog of cyclic AMP, 8-Br cAMP (100 microM), but not by gamma-amino-butyric acid (GABA). Also, the selective antagonist of the benzodiazepine receptors, flumazenil did not modify the effect of diazepam. The diazepam-induced shortening of action potential duration was partially antagonized by the inhibitor of cAMP synthesis carbachol (1 microM) or the blocker of the cAMP-dependent protein kinase A, Rp-cAMP[S] (1 microM). These results indicate that cyclic AMP is involved in the diazepam induced shortening of the action potential duration of the guinea pig papillary muscle. PMID- 10374903 TI - Inhibition of endothelium-dependent vascular relaxation by tetrandrine. AB - The effects of tetrandrine, a Ca2+ antagonist of bis-benzylisoquinoline alkaloid origin, on endothelium-dependent and -independent vascular responsiveness were investigated in perfused rat mesenteric artery. In endothelium-intact preparations pre-contracted with 3 microM phenylephrine and fully relaxed by 0.3 microM acetylcholine tetrandrine caused a rapid transient contraction. In endothelium-denuded preparations, tetrandrine caused only vasorelaxation of phenylephrine-contraction. The biphasic effect of tetrandrine in acetylcholine relaxed preparations could also be mimicked by sequential applications of atropine/tetrandrine or N(G)-nitro-L-arginine-methylester (L-NAME)/tetrandrine, but atropine or L-NAME alone caused only vasoconstriction. This tetrandrine induced transient vasoconstriction was also observed in preparations relaxed with ATP, histamine or thapsigargin (TSG), but not those relaxed with A23187, sodium nitroprusside or nifedipine. The present results suggest that tetrandrine, in addition to its known inhibitory effects on vascular smooth muscle by virtue of its Ca2+ antagonistic actions, also inhibits NO production by the endothelial cells possibly by blockade of Ca2+ release-activated Ca2+ channels. PMID- 10374904 TI - Tissue norepinephrine turnover and cardiovascular responses during intermittent dehydration in the rat. AB - We investigated the central and peripheral sympathetic responses to intermittent dehydration in rats. The norepinephrine (NE) turnover, a biochemical index correlated with noradrenergic neuronal activity, was measured. The modification of blood pressure was also determined by telemetry during the different cycles of dehydration. Dehydration caused a decrease of NE turnover in A2, A5 and A6 nuclei and in peripheral organs. The vasopressinergic level of dehydrated rats decreased in hypophysis and hypothalamus, and increased in plasma. A repeated gradual increase of arterial blood pressure during the first three days of dehydration, followed by a sudden drop when the rats were rehydrated on the fourth day was observed. In conclusion, our study revealed an increase in blood pressure and in central sympathetic activity during dehydration. PMID- 10374905 TI - Protective effect of Phyllanthus fraternus against carbon tetrachloride-induced mitochondrial dysfunction. AB - The effect of carbon tetrachloride administration on liver mitochondrial function and the protective effect of an aqueous extract of Phyllanthus fraternus were studied in rats. The following changes were observed in mitochondria due to the administration of carbon tetrachloride. 1) A decrease in the rate of respiration, respiratory control ratio and P/O ratio using glutamate and malate or succinate as substrates. 2) A decrease in the activities of NADH dehydrogenase (35%), succinate dehydrogenase (76%) and cytochrome c oxidase (51%). The rate of electron transfer through site I, site II and site III was studied independently and found to be significantly decreased. 3) A decrease in the content of cytochrome aa3 (34%). 4) A significant decrease in the levels of phospholipids particularly cardiolipin and a significant increase in the lipid peroxide level was observed. The carbon tetrachloride induced toxicity may be partly due to the lipid peroxidation and partly due to the effect on protein synthesis. Administration of rats with an aqueous extract of P. fraternus prior to carbon tetrachloride administration showed significant protection on the carbon tetrachloride induced mitochondrial dysfunction on all the parameters studied. PMID- 10374906 TI - Relationship of human pancreatic cholesterol esterase gene structure with lipid phenotypes. AB - Pancreatic cholesterol esterase is one of the enzymes that plays a pivotal role in cholesterol absorption. Differences in the genotype of this enzyme could affect the susceptibility of individuals to dyslipidemia and/or cardiovascular disease. We undertook this study to investigate if any correlation exists between restriction fragment length polymorphism in the human pancreatic cholesterol esterase gene and serum lipid levels. DNA from 96 healthy adults was restricted with Stu I, Southern blotted, and probed with cDNA of human pancreatic cholesterol esterase. Results revealed six distinct patterns which were classified as A, B, C, D, E, and F which had a population frequency of 1%, 34.5%, 49%, 12.5%, 1% and 2% respectively. Correlation of the distribution of lipid and lipoprotein levels by pattern and sex revealed a significant interaction between pattern type and HDL (p=0.03) in the most common group (group C) for males. Male patients of pattern C tended to have a lower LDL cholesterol than non-pattern C males (p=0.07); in addition, 80% of all males in the study population with LDL cholesterol under 100 mg/dl were found in pattern C. Thus, the most common Stu I RFLP genotype is associated with a favorable lipid phenotype. This report shows an association between the human pancreatic cholesterol esterase genotype and serum lipid levels. Further analysis of a larger study group with Stu I and alternative polymorphic restriction enzymes is warranted, to confirm this biologically plausible result. PMID- 10374907 TI - Janus kinases and their role in growth and disease. AB - Janus kinases (JAK) play a crucial role in the initial steps of cytokine signaling. Each of the four members (JAK1, JAK2, JAK3, TYK2) of this non-receptor tyrosine kinase family is indispensable for the effects of distinct cytokines. Moreover, recent reports have added to our knowledge on their highly specific functions: JAK3 knockout mice and JAK3 deficient patients cannot signal through the interleukin-2,4,7,9, or 15 receptors and suffer from severe combined immunodeficiency (SCID). JAK1 and JAK2 knockout mice do not survive, their cells again showing distinct patterns of cytokine signaling deficits. At the other end of the spectrum, JAK fusion proteins have been shown to play a role in leukemias. In addition, a new class of JAK-specific inhibitors was described by several groups, the CIS/SOCS/Jab family. This review on the rapidly growing field focuses on JAK function and regulation, and on their emerging role in development and human disease. PMID- 10374908 TI - Effects of glucagon and glucagon-like peptide-1 on glucocorticoid secretion of dispersed rat adrenocortical cells. AB - The effects of glucagon and glucagon-like peptide-1 (GLP-1) on the secretory activity of rat adrenocortical cells have been investigated in vitro. Neither hormones affected basal or agonist-stimulated aldosterone secretion of dispersed rat zona glomerulosa cells or basal corticosterone production of zona fasciculata reticularis (inner) cells. In contrast, glucagon and GLP-1 partially (40%) inhibited ACTH (10(-9) M)-enhanced corticosterone secretion of inner cells, maximal effective concentration being 10(-7) M. The effect of 10(-7) M glucagon or GPL-1 was suppressed by 10(-6) M Des-His1-[Glu9]-glucagon amide (glucagon-A) and exendin-4(3-39) (GPL-1-A), which are selective antagonists of glucagon and GLP-1 receptors, respectively. Glucagon and GLP-1 (10(-7) M) decreased by about 45-50% cyclic-AMP production by dispersed inner adrenocortical cells in response to ACTH (10(-9) M), but not to the adenylate cyclase activator forskolin (10(-5) M). Again this effect was blocked by 10(-6) M glucagon-A or GLP-1-A. The exposure of dispersed inner cells to 10(-7) M glucagon plus GLP-1 completely suppressed corticosterone response to ACTH (10(-9) M). However, they only partially inhibited (by about 65-70%) both corticosterone response to forskolin (10(-5) M) or dibutyryl-cyclic-AMP (10(-5) M) and ACTH (10(-9) M)-enhanced cyclic-AMP production. Quantitative HPLC showed that 10(-7) M glucagon or GLP-1 did not affect ACTH-stimulated pregnenolone production, evoked a slight rise in progesterone and 11-deoxycorticosterone release, and markedly reduced (by about 55%) corticosterone secretion of dispersed inner adrenocortical cells. In light of these findings the following conclusion are drawn: (i) glucagon and GLP-1, via the activation of specific receptors, inhibit glucocorticoid response of rat adrenal cortex to ACTH; and (ii) the mechanism underlying the effect of glucagon and GLP-1 is probably two-fold, and involves both the inhibition of the ACTH induced activation of adenylate cyclase and the impairment of the late steps of glucocorticoid synthesis. PMID- 10374909 TI - Possible involvement of nitroglycerin converting step in nitroglycerin tolerance. AB - Nitroglycerin (GTN) produces a dilation of vascular smooth muscle by releasing NO through a putative GTN-converting step. However, the response to GTN is markedly attenuated after prolonged or repeated exposure, resulting in tolerance. We investigated the mechanisms of GTN tolerance, employing exogenous and endogenous NO in rat aorta. In endothelium-denuded rat aortic strips, the GTN-induced relaxation response was attenuated by preceding exposure to either GTN or sodium nitroprusside (SNP). In contrast, the SNP-induced relaxation response was not affected by this protocol of GTN or SNP preexposure. Preincubation of aortic strips with lipopolysaccharide (LPS) +/- L-arginine for 12 h also caused attenuation of GTN-induced responses such as relaxation, cGMP production and nitrite/nitrate formation. The attenuating effect of LPS abolished in aortic strips co-incubated with LPS and cycloheximide or N(G)-nitro-L-arginine. These results suggest that GTN tolerance is predominantly associated with the reduction of NO release from GTN, which is caused through inhibition of a GTN-converting step due to preceding exposure to NO itself. PMID- 10374910 TI - The gut cytokine balance as a target of lead toxicity. AB - The impact of exposure to lead on gut cytokine gene expression and oral tolerance was analyzed. Oral tolerization with ovalbumin (OVA) increased levels of IL-10 and TGF-beta in gut tissue while IFN-gamma mRNA levels remained unchanged in both autoimmune diabetes prone NOD and normal C57BL/6 mice. This shift towards Th2/Th3 type cytokine gene expression was completely abolished by concomitant treatment with PbCl2 (6 x 0.5 mg/kg) in NOD mice while the cytokine balance in C57BL/6 mice was unaffected. Suppression of Th2/Th3 type cytokine expression was associated with a dampened oral tolerance response to OVA as determined by T cell proliferation assays. We conclude that in autoimmunity prone NOD mice environmental toxicants may disturb immune homeostasis by targeting the gut immune system. PMID- 10374911 TI - Dual intracellular pathways in gonadotropin releasing hormone (GNRH) induced desensitization of luteinizing hormone (LH) secretion. AB - The mechanisms of GnRH-induced desensitization of LH secretion are poorly understood. Protein kinase C (PKC) and protein kinase A (PKA) desensitize some receptors of the 7-membrane type, and the GnRH receptor has consensus phosphorylation sites for PKC in the first and third intracellular loops, and a site for PKA in the first intracellular loop. In the first set of experiments we determined whether synthetic peptides representing the three intracellular loops of the receptor could be phosphorylated in vitro by purified PKC and PKA. As compared with a model substrate peptide for PKC, the third intracellular loop was phosphorylated 74% and the first intracellular loop 21%; PKA-phosphorylated the first intracellular loop peptide 17% as well as a model peptide substrate. In the second set of experiments, we used phorbol 12-myristate 13 acetate (PMA), an established PKC stimulator, and cholera toxin (CTX), established to activate the Gs protein and presumed to activate PKA, to treat cultured rat pituitary cells followed by LH measurements. Treatment with both drugs severely impaired GnRH stimulated LH secretion whereas neither drug alone reduced LH secretion. Dibutyryl cAMP did not duplicate the effects of cholera toxin suggesting that the CTX action could not be explained by an increase in cAMP. These results suggest that more than one intracellular signaling pathway requires activation in order to induce desensitization; one pathway involves PKC and the other involves a pathway stimulated by cholera toxin, presumably Gs protein, which does not involve PKA. PMID- 10374912 TI - Effect of stress on the Schultz-Dale reaction in guinea pig aorta. AB - The smooth muscle of thoracic aorta from guinea pig sensitized with egg albumin (EA) produced an anaphylactic contraction when it was exposed to EA. Experiments were performed to evaluate stress effects on the anaphylactic contraction in guinea pig aortic rings. Two types of stressors were used as immunosuppressor stimuli: physical restraint and shaking of the animals. Both stressors diminished the amplitude of the Schultz-Dale contraction in aortic rings from sensitized guinea pig. The shake stress stimulus interrupted several times during each session induced higher immunosuppression in animals in which the active sensitization and the stress sessions began the same day. Severe restraint stress, prior to active immunization, also suppressed significantly the anaphylactic response. The Schulz-Dale reaction in guinea pig aorta seems to be a valuable technique to study the stress effects on the anaphylactic response. PMID- 10374913 TI - Effect of cysteamine hydrochloride on secretion of growth hormone in male swine. AB - The objective of this study was to determine the effect of cysteamine hydrochloride (CSH) on growth hormone (GH) secretion in male swine. Twelve Poland China x Yorkshire boars, weighing 103.4 +/- 3.0 kg and fitted with indwelling jugular vein catheters, were individually penned in an environmentally controlled room. Boars received i.v. injections of either 0, 25, 50, or 75 mg CSH/kg body weight (BW) at h 0 (n = 3/treatment). Blood samples were collected every 15 min from h 0 to h 4. Serum concentrations of GH were determined by radioimmunoassay. There was an effect of treatment (P < .05) on mean GH concentrations. Mean GH concentrations (ng/ml) were 1.97 +/- .46, 2.24 +/- .59, .91 +/- .06, and .62 +/- .08 for boars receiving 0, 25, 50, and 75 mg CSH/kg BW, respectively. The dose of CSH-mean GH response had a linear (P < .01) component. Cysteamine hydrochloride at the 75 mg/kg BW dose decreased mean GH concentrations (P < .05) compared to the 0 and 25 mg/kg BW groups. The frequency and amplitude of GH pulses were similar (P > .1) among treatments. Overall, GH pulse amplitude was 2.35 +/- .58 ng/ml and GH pulse frequency was .75 +/- .07 pulses/h. Results from this experiment indicate that CSH suppresses circulating GH concentrations in a dose dependent fashion in boars. PMID- 10374914 TI - Pharmacological characterization of dopamine transport in cultured rat astrocytes. AB - The effects of GBR-12909 (selective DA uptake inhibitor), zimelidine (selective 5 HT uptake inhibitor) and nisoxetine (selective NE uptake inhibitor) on the uptake of 30 nM [3H]DA into cultured rat astrocytes were examined. [3H]DA uptake was inhibited by approximately 50% by GBR-12909 or zimelidine in a concentration dependent manner (100 nM to approximately 10 microM). Furthermore, the inhibition curves of GBR-12909 were biphasic, and uptake was completely inhibited by a high concentration of GBR-12909 (100 microM). [3H]DA uptake was also inhibited by approximately 50% by nisoxetine in a concentration-dependent manner (0.1 to approximately 100 nM), and nisoxetine was more potent than GBR-12909 or zimelidine. The inhibitory potencies were in the order nisoxetine > GBR-12909 > zimelidine. The uptake of [3H]DA under Na+-free conditions was approximately 50% of that under normal conditions. Thus, DA was taken up by both Na+-dependent and Na+-independent mechanisms. Nisoxetine (100 nM), zimelidine (100 microM) and GBR 12909 (10 microM) inhibited [3H]DA uptake into astrocytes only in the presence of Na+. On the other hand, this uptake was completely inhibited by a high concentration of GBR-12909 (100 microM) in the absence of Na+. The present data suggest that the Na+-dependent uptake of [3H]DA in cultured rat astrocytes may occur in the NE uptake system. Furthermore, astrocytes express the extraneuronal monoamine transporter (uptake2), which is an Na+-independent system, and this transporter is involved in the inactivation of centrally released DA. PMID- 10374915 TI - Effect of somatostatin on beta-endorphin release in rat experimental chronic inflammation. AB - The aim of the present study was to investigate the effect of somatostatin administration in arthritic rats. Inflammation was induced by daily interplantar injection of 100 microl of Freund's complete adjuvant into the left hind paw of the rat. Arthritis developed 20 days following the first injection and was stable in the inoculate paw. Arthritic rats were treated interplantarly with somatostatin (5 or 10 microg) or with indomethacin (100 microg) daily for 14 days. Inflammatory response was studied at 12 h, 7 and 14 days following drug administration. The effect of somatostatin was determined by local (into popliteal lymph nodes) and systemic production of beta-endorphin. Our results showed that somatostatin treatment significantly increased beta-endorphin levels in the blood and lymphocytes from popliteal lymph nodes. Greater efficiency was seen when 5 microg instead of 10 microg of somatostatin was used. A significant decrease of absolute leukocytosis was observed at the 14th day following somatostatin administration. Moreover, a significant reduction of plasmatic beta globulins at 12 h and the 7th day and of plasmatic alpha2-globulins at the 14th day was observed after the beginning of somatostatin treatment. PMID- 10374917 TI - In vivo survival of selected murine carrier red blood cells after separation by density gradients or aqueous polymer two-phase systems. AB - In order to explore possibilities of using erythrocytes as carrier systems for delivery of pharmacological agents, we have studied the in vivo survival of murine carrier red blood cell populations enriched in young or old cells. Hypotonic-isotonic dialysis has been used to modify the cells as carrier systems and Percoll/albumin density gradients or counter-current distribution in aqueous polymer two-phase systems to separate them according to age. Hypotonic-isotonic dialysis produces a decrease in the red blood cell populations in vivo survival rate (from 9.5 to 7.8 days). Among the cells modified as carriers, the enriched young red blood cell populations show a higher in vivo survival (half-life 6.5 7.4 days) than populations made up of predominantly old red blood cells (half life 4.7-6.2 days). Half-life of young or old circulating red blood cells was approximately one day longer when these cells were separated by counter-current distribution rather than by Percoll density gradients. Based on these results, hypotonic-isotonic dialysis of whole and enriched young or old red blood cell populations, with higher or lower survival rates, can be considered as a useful tool for modification of these cells as carriers. The final outcome of such changes can be translated into better control of plasma drug delivery during therapy. PMID- 10374916 TI - Liver mRNA expression of IGF-I and IGFBPs in adult undernourished diabetic rats. AB - To investigate the role played by factors other than GH, such as nutrients and insulin, on IGF-I secretion, adult male rats of 200 g.b.w. were food-restricted for 7 days and then made diabetic by streptozotocin administration (UD). Different groups of UD rats were submitted to the following four day treatments: left untreated (UD), refed (UD+R), treated with insulin (UD+I), or a combination of both refeeding and insulin (UD+R+I). Serum concentration of IGF-I and liver mRNA expression of IGF-I, IGF-binding proteins and GH receptor were measured. Insulin treatment alone partially recovered liver IGF-I and IGFBPs mRNA expression, while refeeding alone had no effect. Only a combination of both insulin and refeeding recovered both parameters. Contrary to the results obtained with a longer period of recovery, these experiments show that serum and mRNA expression of IGF-I and IGFBPs in adult undernourished diabetic rats can be restored by insulin and nutrients administration with no prior restoration of serum and pituitary GH to control values and no compensatory changes in GH receptor gene expression. PMID- 10374918 TI - Activity of renal 11betahydroxysteroid dehydrogenase 2 (11betaHSD2) in stressed animals. AB - The enzyme 11betaHSD2 protects the non-selective mineralocorticoid receptor from occupation by glucocorticoids in aldosterone target tissues. We studied the effect of stress elicited by intubation with a rubber catheter and administration of 10 ml of 0.45% NaCl (G3), of 10 ml of 200 mM HCl (G4) or intubation alone (G2) on the kinetics of the renal enzyme compared with untreated rats (G1). Microsomes were incubated with increasing masses of 3H corticosterone and 400 microM NAD at pH=7.4 during 5 minutes. Samples were extracted with ethyl acetate and analyzed by TLC. Results for n=4: Vmax for G1, 4.82 +/- 0.67. G2, 10.04 +/- 0.16***. G3, 9.16 +/- 0.74**. G4, 10.19 +/- 0.79*** pmoles/min/mg prot. Km for G1, 22.37 +/- 2.42. G2, 50.72 +/- 7.05*. G3, 55.25 +/- 8.37**. G4, 27.40 +/- 3.20 nM. (***p<0.001, **p<0.01 and *p<0.05 vs G1). All treatments increased Vmax. Intubation alone and gavage with 0.45% NaCl, but not with 200 mM HCl, increased Km. Taking together, the results could reflect a way to prevent occupation of type I receptors by increased levels of circulating glucocorticoids due to stressful situations. This protection seems more efficient under acidotic conditions causing--in addition to an increased Vmax--a low Km for the enzyme. PMID- 10374919 TI - Age-related differences in serum melatonin and pineal NAT activity and in the response of rat pineal to a 50-Hz magnetic field. AB - In a previous study we have shown that exposure to a 50-Hz sinusoidal magnetic field decreased serum melatonin concentration and pineal enzyme activities in young rats (9 weeks). In the present study we looked for the effect of a magnetic field of 100 microT on serum melatonin and pineal NAT activity in aged rats and compared them to young rats. We hypothesized that aging may change sensitivity of rats to a magnetic field. Two groups of Wistar male rats [aged rats (23 months) and young rats (9 weeks)] were exposed to 50-Hz magnetic fields of 100 microT for one week (18h/day). The animals were kept under a standard 12:12 light: dark cycle with a temperature of 25 degrees C and a relative humidity of 45 to 50%. Control (sham-exposed) animals were kept in a similar environment but without exposure to a magnetic field. The animals were sacrificed under red dim light. Serum melatonin concentration and pineal N-acetyltransferase (NAT) and hydroxyindole-O-methyltransferase (HIOMT) activities were studied. Our results showed that sinusoidal magnetic fields altered the production of melatonin (28% decrease; P <0.05) through an inhibition of pineal NAT activity (52% decrease; P <0.05) in the young rats whereas no effect was observed in aged ones. On the other hand, when comparing data from control animals between young and aged rats, we observed that serum melatonin level and NAT activity, but not HIOMT activity, decreased in aged rats (decrease by about 38% and 36% respectively). Our data strongly suggest that old rats are insensitive to the magnetic field. PMID- 10374920 TI - Fluctuation of serum leptin level in rats after ovariectomy and the influence of estrogen supplement. AB - In order to understand the mechanism of increasing body fat in perimenopausal and postmenopausal women, an ovariectomy-induced obesity model was used to study the role of leptin. In this investigation, a long-term study lasted for 13 weeks was conducted to monitoring the change of serum leptin level in rats after the loss of estrogen, and also to examine the influence of estrogen replacement. The results showed that three weeks after the removal of ovaries the body weight of Ovx rats was already significantly higher than the other two groups, and continued to gain more weight thereafter. Accompanying with the significant weight gain was the changes in the serum leptin levels. The leptin concentration declined gradually during the first half of experimental period, dropping down to an almost undetectable level at week 7 (0.216+/-0.132 ng/ml). Subsequently, its concentration began to elevate, and by the end of experiment leptin level was significantly higher (3.182+/-0.936 ng/ml) than the value before the operation (0.818+/-0.242 ng/ml). This fluctuation of serum leptin level caused by ovariectomy was eliminated by the replacement of estrogen. The present data indicate that ovariectomy-induced weight gain is caused by the early drop in leptin level. The later rise in leptin production is connected to the increased body weight probably originated from a reduced sensitivity in leptin signal. PMID- 10374921 TI - Endothelium-dependent effect of bradykinin and ATP on rabbit ventricular myocytes. AB - We have investigated the effects of bradykinin (BK) and ATP on Ca2+ transient induced by electrical-field stimulation in freshly isolated rabbit ventricular myocytes, in the presence or absence of rabbit aortic endothelial cells. BK and ATP induced an increase in intracellular Ca2+ concentration ([Ca2+]i) in the endothelial cells, but had no significant effect on Ca2+ transient in electrical field stimulated ventricular myocytes. In the presence of cultured endothelial cells, the amplitude of Ca2+ transient induced by electrical stimulation in ventricular myocytes was decreased. BK and ATP further reduced the amplitude of Ca2+ transient induced by electrical stimulation in ventricular myocytes. These data show that BK and ATP have endothelium-dependent depressing effects on ventricular myocytes and indicate that substances released from endothelial cells in response to BK and ATP stimulation can modulate ventricular myocytes excitation-contraction coupling. However, identification of the cardioactive mediators produced by the endothelial cells requires further study. PMID- 10374922 TI - The effects of heparin-binding epidermal growth factor-like growth factor on preimplantation-embryo development and implantation in the rat. AB - This study examined the effects of heparin-binding epidermal growth factor-like growth factor (HB-EGF) on preimplantation-embryo development and initiation of implantation in the rat. In vitro studies showed that HB-EGF improved the development of 8-cell embryos to the blastocyst stage in a concentration dependent manner, and the growth factor had no effect on the cell number of the blastocyst developed. Intraluminal injection of an anti-HB-EGF antiserum into the uterine horns at 0600 h on day 5 of pregnancy decreased the number of implantation sites (blue dye reaction) at 0200 h on day 6. Intraluminal injection of 20 microl of HB-EGF solution (10 or 100 ng/ml) into each uterine horn induced implantation in about half of the ovariectomized progesterone-treated delayed implanting rats, and the number of implantation sites per rat increased dose dependently. These results suggest that HB-EGF is involved in the preimplantation embryo development and initiation of implantation in the rat. PMID- 10374923 TI - Protein kinase C promotes spontaneous relaxation of streptolysin-O-permeabilized smooth muscle cells from the guinea-pig stomach. AB - Isolated single smooth muscle cells from the fundus of a guinea-pig stomach were permeabilized by use of streptolysin-O (0.5 U/ml). Most of the permeabilized cells responded to 0.6 microM Ca2+, but not to 0.2 microM Ca2+, with a resulting maximal cell shortening to approximately 71% of the resting cell length. These cells were relaxed again by washing with the Ca2+-free solution (2.5 nM free Ca2+) for 3-5 min. Addition of 10 microM acetylcholine (ACh) resulted in both a marked decrease in the concentration of Ca2+ required to trigger a threshold response and an increase in the maximal cell shortening, indicating that the cells retained the muscarinic receptor function. When the cell treated with a protein kinase C (PKC) inhibitor, K-252b (1 microM), for 3 min was exposed to 10 microM ACh in the presence of K-252b, the cell shortened within 2 min with a maximal cell shortening. When the cell shortening was induced by 10 microM ACh plus 1 microM Ca2+ in the presence of K-252b (1 microM) or more selective PKC inhibitors, such as calphostin C (1 microM) or PKC pseudosubstrate peptide (100 microM), the extension of the shortened cells, by washing with the Ca2+-free solution, was significantly inhibited. In contrast, K-252b (1 microM) did not inhibit the relaxation of Ca2+-induced shortened cells. These results suggest that the receptor-mediated activation of PKC in the process of ACh-induced cell shortening plays a role in the subsequent relaxation of the shortened cells. PMID- 10374924 TI - Identification of linear surface epitopes on the guinea pig sperm membrane protein PH-20. AB - The sperm plasma membrane protein PH-20 has previously been shown to be an effective immunogen for protection against fertilization in guinea pigs. To identify immunodominant regions on gpPH-20 that may be related to this contraceptive effect, we used several high-titer immune sera obtained from animals rendered infertile by gpPH-20 injections to screen a set of overlapping peptides that cover the entire 494-residue sequence. Multiple clusters of peptide sequences exhibited specific reactivity. Some of these sequences were then constructed as octameric synthetic peptides and tested for immunogenicity in female guinea pigs. Our results indicated two regions (res. 94-119 and res. 424 444) to be highly immunogenic and both are surface accessible when native gpPH-20 is in solution or anchored on sperm surface. Both anti-peptide antibodies are specific for gpPH-20 and one of them inhibited hyaluronidase activity partially. These monospecific antibodies should be useful probes for further molecular definition of gpPH-20 structure-function relationships. PMID- 10374925 TI - Folic acid intestinal absorption in newborn rats at 21 day postpartum: effects of maternal ethanol consumption. AB - This study was designed to examine the effects of prenatal and postnatal exposure of ethanol in the in vivo absorption of free folic acid in the small intestine in pups rats at the 21st day after birth. The rats were accustomed to increasing amounts of ethanol (5 to 20%, vol/vol) in tap water for 1 month. During pregnancy and suckling period, ethanol-fed dams were assigned again to ethanol 20% in drinking water. Two sets of experiments were performed. In the first set, jejunal free folic acid absorption in control group and litters nursed by dams receiving ethanol showed a gradual increase along with the increase of perfusion time at all the assayed concentrations. In general, in litters of ethanol-fed dams, jejunal free folic acid absorption expressed as nmol/intestinal surface, nmol/g tissue wet weight and nmol/g tissue dry weight were higher than in control animals. In the second set of experiments, in distal ileum loops, free folic acid absorption did not occur in control pups, but appeared in litters exposed to ethanol. Milk folic acid levels are significantly decreased in ethanol-treated dams. However, only a slight decrease in the serum folic acid levels occurs in litters of ethanol-fed dams. In conclusion, the results obtained in the present work suggested a different pattern of free folic acid absorption in distal ileum for the two groups. The exposure of rats to ethanol during the pregnancy and suckling period, can affect postnatal development of intestinal functions and could play a role in the genesis of malnutrition observed in the infant. PMID- 10374926 TI - Opioid binding profiles of new hydrazone, oxime, carbazone and semicarbazone derivatives of 14-alkoxymorphinans. AB - Several hydrazone, oxime, carbazone and semicarbazone derivatives of 14 alkoxycodeinones and 14-alkoxydihydrocodeinones were synthesised [1] and characterised in in vitro radioligand binding assays in rat brain membrane preparations. The tested compounds show the highest affinity for the mu opioid binding sites and most of them have agonist character. Subtype analysis of the binding shows mu2 specificity. However, some of these ligands are able to block partially (40-60%) the high affinity (putative mu1) opioid binding sites while all of them act as reversible ligands at the low affinity (putative mu2) sites. PMID- 10374927 TI - Peripheral orphanin FQ/nociceptin analgesia in the mouse. AB - Orphanin FQ/Nociceptin (OFQ/N) administered peripherally was an effective analgesic in the tailflick test in mice (ED50 16.3 microg). It had a peak effect at 5 min and lasted up to 30 min. The kappa3 analgesic naloxone benzoylhydrazone was also active peripherally (ED50 3.8 microg). The analgesic actions of both agents were blocked by naloxone. Neither OFQ/N(1-11) nor OFQ/N(1-7) had appreciable peripheral activity. Antisense mapping both compounds against the murine orphan opioid receptor (KOR-3) confirmed the importance of this clone in their actions. Antisense probes targeting the second and third coding exons significantly lowered the analgesic effects of both compounds. However, the antisense targeting the first coding exon blocked only the actions of OFQ/N and not kappa3 analgesia. PMID- 10374928 TI - Presence of opioid receptor-like (ORL1) receptor mRNA splice variants in peripheral sensory and sympathetic neuronal ganglia. AB - The expression of ORL1 receptor mRNA splice variants is determined in peripheral sensory and sympathetic ganglia and compared to mRNA expression for the three classic opioid receptor subtypes (mu, delta, and kappa) using the method of reverse transcription-polymerase chain reaction. ORL1, mu, delta and kappa receptor subtype mRNAs are present in human dorsal root ganglia (DRG) and trigeminal ganglia and rat DRG. ORL1, mu and delta receptor subtype mRNAs are present in rat superior cervical ganglia and only ORL1 and delta receptor mRNAs are present in rat lumbar sympathetic ganglia. Both the ORL1 mRNA splice variants are present in sensory and sympathetic ganglia, however, expression of the shorter ORL1 receptor mRNA dominates over expression of the longer splice variant in rat brain and DRG, whereas, expression of the longer splice variant is dominant in sympathetic ganglia. PMID- 10374929 TI - AgNOR proteins from morphologically intact isolated nucleoli. AB - AgNOR staining has been proposed as a useful tool for the diagnosis and prognosis of cancer. The AgNOR proteins, however, have not yet been clearly identified and characterized, possibly due to the partial character of the results obtained when studying the proteins extracted from altered nucleoli isolated by "standard" methods. In the present study, we analysed, on western blots, the AgNOR staining profiles obtained with protein extracts from Ehrlich tumor cell nucleoli isolated by a recent procedure that preserves the nucleolar ultrastructure. In addition to the well-known C23 and B23 protein bands, we readily detected an extra band at approximately 125 Kda. By immunoblotting, we showed that this polypeptide may be related to the nucleolar phosphoprotein pp135 evidenced in rat-cell nucleoli. By immunoelectron microscopy, we detected this protein in the dense fibrillar component and fibrillar center of the nucleoli as well as the coiled bodies. The distribution coincides with the cytochemical AgNOR staining pattern obtained at the ultrastructural level. PMID- 10374930 TI - Two distinct pathways for refilling Ca2+ stores in permeabilized bovine trachealis muscle. AB - Calcium entry from extracellular space to acetylcholine (ACh)-sensitive internal stores was investigated in beta-escin permeabilized bovine tracheal smooth muscle. Cyclopiazonic acid (CPA), a selective inhibitor of the sarcoplasmic reticulum (SR) calcium pump, and nifedipine, both inhibited the refilling, and inhibition was larger when these compounds were used simultaneously. BayK 8644 enhanced the refilling and completely reversed the inhibition induced by cyclopiazonic acid. In pCa 7 solution containing CPA, there was a spontaneous time-dependent decrease of ACh-induced transient contraction. In the presence of nifedipine or verapamil in the incubation solution reduced this time-dependent decrease in contractile responses to ACh stimulation, suggesting that these calcium-entry blockers decreased calcium leakage from internal stores to the extracellular space. These results suggest that in addition to the active calcium uptake in the SR, another pathway controlled by an L-type like calcium channel (dihydropyridine-sensitive) may exist between the extracellular compartment and the lumen of the SR in airway smooth muscle, and contributes significantly to the loading of ACh-sensitive calcium stores. PMID- 10374931 TI - Effects of TH-142177 on angiotensin II-induced proliferation, migration and intracellular signaling in vascular smooth muscle cells and on neointimal thickening after balloon injury. AB - We investigated the effects of TH-142177 (N-n-butyl-N-[2'-(1-H-tetrazole-5-yl) biphenyl-4-yl]-methyl-(N-carboxy methyl-benzylamino)-acetamide), a novel selective antagonist of angiotensin II type 1-receptor (AT1-R) on angiotensin II (AII)-induced proliferation and migration of vascular smooth muscle cells (VSMC), and on neointimal formation in the rat carotid artery after balloon injury, and on the intracellular signaling by the stimulation of AT1-R. High affinity AII receptor sites were detected in rat VSMC by the use of [125I]Sar1,Ile8-AII. TH 142177 and losartan competed with [125I]Sar1,Ile8-AII for the binding sites in VSMC in a monophasic manner, although PD123177, a selective antagonist of angiotensin II type 2-receptor (AT2-R), had little inhibitory effect, demonstrating the predominant existence of AT1-R in rat VSMC. TH-142177 prevented AII-induced DNA synthesis and migration, with a significant inhibition of 74 and 55%, respectively, at the concentration of 100 nM. AII-induced activation of p21ras, mitogen-activated protein kinase (p42MAPK and p44MAPK), and protein kinase C was significantly (50-78%) inhibited by TH-142177 (100 nM), suggesting that the activation of these enzymes is mediated through the stimulation of AT1 R. Balloon-injured left carotid arteries in rats showed extensive neointimal thickening, and TH-142177 (3 mg/kg) brought out a marked decrease in the neointimal thickening after balloon injury. In conclusion, TH-142177 inhibited AII-induced proliferation and migration of rat VSMC and neointimal formation in the carotid artery after balloon injury, and these effects may be related, in part, to the suppression of ras, p42MAPK and p44MAPK, and protein kinase C activities through the blockade of AT1-R. Thus, TH-142177 may have therapeutic potential for the treatment of vascular diseases such as atherosclerosis and restenosis. PMID- 10374932 TI - The beta2-adrenergic modulator celiprolol reduces insulin resistance in obese Zucker rats. AB - Essential hypertension is associated with an increased incidence of insulin resistance of skeletal muscle glucose transport. The present study determined if celiprolol, an antihypertensive agent with selective beta1-adrenoceptor antagonist and additional beta2-agonistic properties, administered by gavage either acutely (3 hr) or chronically (14 d), had a direct effect on improving glucose tolerance and insulin-stimulated glucose transport activity (using 2 deoxyglucose (2-DG) uptake) in isolated epitrochlearis muscles of the insulin resistant obese Zucker rat. The effects of a selective beta1-blocker, metoprolol, were also assessed. Acute administration of celiprolol, but not metoprolol, increased insulin-stimulated 2-DG uptake in muscle by 22% (p<0.05). Chronic celiprolol treatment significantly lowered fasting plasma insulin (22%) and free fatty acids (40%) in comparison to obese control values. Moreover, chronic celiprolol administration decreased the glucose-insulin index (calculated as the product of the glucose and insulin areas under the curve during an oral glucose tolerance test), by 32% (p<0.05) compared to obese controls, indicating that peripheral insulin action was increased. Indeed, insulin-stimulated skeletal muscle 2-DG uptake was enhanced by 49% (p<0.05) in these celiprolol-treated obese animals. Metoprolol was without significant effect on any of these variables following chronic administration. These findings indicate that, in this animal model of insulin resistance, the beta1-antagonist/beta2-agonist celiprolol has a specific effect of improving insulin-stimulated skeletal muscle glucose transport that is independent of any hemodynamic alterations. PMID- 10374933 TI - N-(4-hydroxyphenyl) retinamide inhibits cystogenesis by polycystic epithelial cell lines in vitro. AB - Primary tubular epithelial cells develop spherical monolayered cysts when cultured in collagenI matrix, a model that has been used to study the mechanism of cystogenesis. In an attempt to block cystogenesis, we have evaluated the effect of N-(4-hydroxyphenyl) retinamide (HPR), a synthetic derivative of retinoic acid, on both formation and growth of cysts in a human model of polycystic kidney cells. Number, dimension and submicroscopical characteristics of cysts were evaluated after 2 and 4 weeks from treatment with HPR. A marked inhibitory effect of HPR on cystogenesis was found at concentration of 1 microM, while a complete block was observed at concentration between 5 and 10 microM. Furthermore, treatment with HPR of already formed cysts resulted in their disruption. HPR at 10 microM also induced apoptosis of several tubular epithelial cell models suggesting a correlation between the two phenomena. Taken together these observations demonstrate that HPR blocks cystogenesis by polycystic kidney cells "in vitro" and that it also reverts the fate of already formed cysts. Apoptosis may be the mechanism which mediates the inhibitory effect on cystogenesis in this model. PMID- 10374934 TI - Initiation of ionizing radiation-induced apoptosis: DNA damage-mediated or does ceramide have a role? AB - PURPOSE: To critically review the data supporting membrane damage-induced increases in ceramide levels as the primary initiator of ionizing radiation induced apoptosis and to point out that there is compelling evidence supporting the involvement of DNA damage in this process. CONCLUSIONS: There is now a significant literature suggesting that irradiation of cells can quickly lead to a modest, transitory increase in the level of the putative second messenger ceramide. These results have been used to support the views that membrane damage is the primary trigger for radiation-induced apoptosis and that DNA damage is irrelevant to this process. It is argued, however, that the data are inadequate to support such conclusions because it is questionable whether the induced levels of ceramide are toxic and because the ceramide hypothesis cannot convincingly explain the delayed apoptosis, dependent on events such as mitosis, that is shown by many cell lines. In contrast, it is suggested that the sensitivity of some cell types to the induction of apoptosis by DNA-targeted radiation damage, the relationship between p53 status and radiation response, and the influence of enzymatic DNA repair capability on susceptibility to apoptosis, argue strongly that DNA damage is relevant to the triggering of apoptosis. PMID- 10374935 TI - Delayed expression of hpS2 and prolonged expression of CIP1/WAF1/SDI1 in human tumour cells irradiated with X-rays, fission neutrons or 1 GeV/nucleon Fe ions. AB - PURPOSE: Differences in gene expression underlie the phenotypic differences between irradiated and unirradiated cells. The goal was to identify late transcribed genes following irradiations differing in quality, and to determine the RBE of 1 GeV/n Fe ions. MATERIALS AND METHODS: Clonogenic assay was used to determine the RBE of Fe ions. Differential hybridization to cDNA target clones was used to detect differences in expression of corresponding genes in mRNA samples isolated from MCF7 cells irradiated with iso-survival doses of Fe ions (0 or 2.5 Gy) or fission neutrons (0 or 1.2 Gy) 7 days earlier. Northern analysis was used to confirm differential expression of cDNA-specific mRNA and to examine expression kinetics up to 2 weeks after irradiation. RESULTS: Fe ion RBE values were between 2.2 and 2.6 in the lines examined. Two of 17 differentially expressed cDNA clones were characterized. hpS2 mRNA was elevated from 1 to 14 days after irradiation, whereas CIP1/WAF1/SDI1 remained elevated from 3 h to 14 days after irradiation. Induction of hpS2 mRNA by irradiation was independent of p53, whereas induction of CIP1/WAF1/SDI1 was observed only in wild-type p53 lines. CONCLUSIONS: A set of coordinately regulated genes, some of which are independent of p53, is associated with change in gene expression during the first 2 weeks post-irradiation. PMID- 10374936 TI - Selection and sequencing of interchromosomal rearrangements from gamma-irradiated normal human fibroblasts. AB - PURPOSE: To determine the sequences that flank sites of interchromosomal DNA rearrangements and to determine the relative frequency of inter- and intrachromosomal rearrangements induced by 30 Gy gamma-irradiation in a region 5' from exon I of the c-myc gene in normal human fibroblasts (IMR-90). MATERIALS AND METHODS: A modification of an inverse polymerase chain reaction (PCR) procedure, developed previously to detect rearrangements, was used. Inverse PCR products were re-amplified using primers designed to determine whether the product was a result of an inter- or intrachromosomal rearrangement. Possible interchromosomal rearrangements were then sequenced. RESULTS AND CONCLUSIONS: Four of 12 different products analyzed were potentially derived from interchromosomal rearrangements, while the remainder derived from intrachromosomal rearrangements. For three of the potential interchromosomal rearrangements, the sequence recombining with c myc was unidentified, while in the other case the sequence was homologous to an L1 element. The frequencies of inter- and intrachromosomal rearrangements induced by 30 Gy gamma-irradiation in a 2 kbp region flanking the c-myc gene of IMR-90 cells were calculated to be at least 1.6x10(-4) and 3.3x10(-4) respectively. No clear association between sequence context and sites of radiation-induced rearrangement was found; however, two of the four sequenced rearrangements involved breakpoints in the 5'-flanking region of c-myc that occurred immediately after the sequence AAAGG. PMID- 10374937 TI - DNA-PK activation by ionizing radiation-induced DNA single-strand breaks. AB - PURPOSE: To assess the ability of 60Co gamma-radiation-induced plasmid DNA single strand breaks (gamma-ssb) to activate the DNA-dependent protein kinase (DNA-PK) in vitro. MATERIALS AND METHODS: Plasmid DNA was gamma-irradiated under aerobic conditions to yield 0-6 gamma-ssb and <0.1 double-strand breaks (dsb) per plasmid molecule. The irradiated DNA was used to stimulate DNA-PK in crude HF19 fibroblast nuclear extracts and/or purified HeLa cell DNA-PK protein, and the activation compared with that obtained with a single enzymatically generated plasmid DNA ssb (GpII endonuclease) or dsb (EcoRI endonuclease). RESULTS: Gamma Irradiated plasmid DNA activates DNA-PK in both crude and purified preparations and the kinase activity increases linearly with dose. As significant DNA-PK activation was detectable using irradiated plasmids which contain <0.1 dsb/molecule, it was concluded that this activation is due to gamma-ssb. However, using purified DNA-PK, this activation is relatively weak as approximately 3 approximately-ssb is equivalent to one GpII-generated DNA ssb or one end of an EcoRI-generated dsb in DNA-PK assays. CONCLUSIONS: As gamma-ssb are in a approximately 20-fold excess of approximately-dsb in vivo for low LET radiation, gamma-ssb may contribute significantly to DNA-PK signalling of gamma-radiation induced DNA damage in vivo. PMID- 10374938 TI - Radiation-induced DNA double-strand breaks and rejoining in malignant glioma cells. AB - PURPOSE: This study was performed to standardize experimental conditions for the quantification by pulsed-field gel electrophoresis (PFGE) of radiation-induced DNA double-strand breaks (dsb) and rejoining in a human malignant brain tumour xenograft model. MATERIALS AND METHODS: Because no correlation was found between DNA dsb induction or rejoining and clinical radiation response for six fresh glioblastoma (GBM) specimens, assay conditions were examined in detail. SF-767 human GBM cells were implanted into the flanks of athymic mice. Resulting tumours were irradiated in vivo, dissociated mechanically or using an enzyme cocktail, and assayed for DNA dsb induction and repair. In other experiments, excised tumour portions were irradiated and allowed to repair either as chunks (>50 mm3 pieces), as minced tumour (approximately 1 mm3 pieces), or as single-cell suspensions. Finally, the effect of holding excised tumours in vitro for times of up to 72 h before irradiation and assay for DNA dsb and cell survival was studied. RESULTS AND CONCLUSIONS: The method of tumour dissociation had no effect on results; however, both the configuration of specimens during irradiation and the in vitro maintenance time markedly affected the experimental outcome. Chunks irradiated in vitro had DNA dsb results that were very similar to those obtained when tumours were irradiated in situ, while minced pieces or single-cell suspensions resulted in steeper dose-response curves. When tumour chunks were maintained at 4 degrees C in medium, DNA dsb induction was not affected for 24 h and DNA dsb rejoining remained constant for 48 h but then decreased. Cell survival, however, decreased continually during the 72 h in vitro maintenance time. PMID- 10374939 TI - Modification of DNA bases by photosensitized one-electron oxidation. AB - PURPOSE: To determine the distribution of base damage within isolated DNA upon oxidation by three type I photosensitizers in aerated aqueous solution. MATERIALS AND METHODS: Aqueous solutions of DNA were exposed to UVA in the presence of riboflavin, benzophenone or menadione. Then, eight modified nucleobases were measured, using HPLC-EC, GC-MS or HPLC with fluorescence detection. RESULTS: The three photosensitizers led to a predominant degradation of guanine bases within DNA. The relative yield of the three main guanine degradation products measured in DNA was similar with the three sensitizers. 8-OxodAdo was also produced in an almost constant yield with respect to its guanine analogue. The yield of oxidized pyrimidines was lower and was found to depend on the photosensitizer used. The results were compared with the yield of photosensitization-induced degradation of the 2'-deoxyribonucleosides. CONCLUSION: The favoured photosensitized degradation of guanine within DNA may be explained by its lower oxidation potential with respect to that of the other bases, together with the occurrence of charge transfer through DNA. The base modification pattern determined in the present work is different from that obtained upon reaction of hydroxyl radicals. Under the latter conditions, pyrimidine oxidation products were generated more efficiently than by photosensitized one-electron oxidation. PMID- 10374940 TI - Impaired p53-mediated DNA damage response, cell-cycle disturbance and chromosome aberrations in Nijmegen breakage syndrome lymphoblastoid cell lines. AB - PURPOSE: To investigate the p53 ionizing radiation-induced response, G1/S cell cycle block and cytogenetic damage in Nijmegen breakage syndrome (NBS) cells characterized by different haplotypes. MATERIAL AND METHODS: Lymphoblastoid cell lines derived from three normals, five NBS and two ataxia telangiectasia (AT) individuals were treated with moderate doses of X-rays and changes in the p53 response were studied by dose-response and time-course experiments. Multiparametric flow cytometry analysis of bromodesoxyuridine-incorporated cells was carried out to analyse G1/S checkpoint alterations. Cytogenetic damage induced by 2 Gy radiation was assessed in cells harvested 28 h later. RESULTS: Comparison of mean values of p53 accumulation in NBS, AT and control cells indicated that protein induction in NBS cells was between normal and AT cells. Cell-cycle experiments showed a markedly reduced S-phase fraction in irradiated samples of normal cell lines, while NBS, and particularly AT cells, showed less reduction in S-phase fraction. Irrespective of differences in p53 induction and G1/S block, chromatid-type aberrations were induced at a comparable level in both syndromes, while being almost absent in normal cells. CONCLUSIONS: The data suggested that failure of NBS cells to initiate cell-cycle delay cannot account alone for their extreme sensitivity to radiation. PMID- 10374941 TI - Estimate of the frequency of true incomplete exchanges in human lymphocytes exposed to 1 GeV/u Fe ions in vitro. AB - PURPOSE: To study the frequency of true incomplete exchanges induced by high-LET radiation. MATERIALS AND METHODS: Human lymphocytes were exposed to 1 GeV/u Fe ions (LET = 140 keV/microm). Chromosome aberrations were analysed by a fluorescence in situ hybridization using a combination of whole-chromosome specific probes and human telomere probes. Chromosomes 1, 3 and 4 were investigated. RESULTS: The percentage of incomplete exchanges was between 23 and 29% if telomere signals were not considered. The percentage decreased to approximately 10% after ruling out false incomplete exchanges containing telomere signals. The final estimation of true incomplete exchanges was <10%. CONCLUSION: Within a degree of uncertainty, the percentage of true incomplete exchanges in 1 GeV/u Fe ion-irradiated human lymphocytes was similar to that induced by gamma rays. PMID- 10374942 TI - Chromosomal instability in haemopoietic cells of the foetus, mother and offspring after in utero irradiation of the CBA/Ca mouse. AB - PURPOSE: The present study was conducted to test the susceptibility of the mouse foetus to transmit chromosomal instability to the haemopoietic stem cells of offspring after in utero X-or plutonium-239-irradiation. MATERIALS AND METHODS: Pregnant CBA/Ca-mice were injected with 80 kBq/kg 239Pu or X-irradiated with 1 Gy X-rays on days 13 or 14 of gestation. CFU-A cultures were grown from haemopoietic stem cells sampled from foetal liver and the bone marrow from the offspring and from the mother. Non-clonal, unstable chromosomal aberrations were scored in metaphases from individual stem cell colonies. RESULTS: The relative excess (RE) of unstable chromosomal aberrations in foetal liver cells irradiated with 1 Gy X rays increased from 1.6 at day 2 up to 2.7 at day 4 after irradiation. In the bone marrow cells from the mother, this value was 1.8 (average from cells sampled at days 3 and 14 after irradiation). After injection of the pregnant mice with 235Pu, the yield of unstable chromosomal aberrations per cell was 0.14+/-0.03 (RE approximately 10) in descendants of bone marrow cells from the mother, 0.11+/ 0.02 (RE = 10) in descendants of foetal liver cells and 0.16+/-0.05 (RE = 10) in descendants of bone marrow cells from the offspring. CONCLUSIONS: From the numerical analysis of non-clonal, unstable aberrations in haemopoietic cells from the foetus, the mother and the offspring after in utero irradiation, it was concluded that in utero irradiation of the CBA/Ca mouse was not more efficient in inducing chromosomal instability in the offspring than in the foetus or the mother. All three cell populations exhibited a similar degree of unstable aberrations, both in terms of the absolute numbers of non-clonal aberrations and in terms of relative excess compared with unexposed controls. PMID- 10374943 TI - Effects of keratinocyte growth factor in the squamous epithelium of the upper aerodigestive tract of normal and irradiated mice. AB - PURPOSE: To investigate the effects of keratinocyte growth factor (KGF) on the structure of the stratified squamous epithelium of the tongue, buccal mucosa and oesophagus of normal and irradiated mice. MATERIALS AND METHODS: Female BDF1 mice were exposed to total body irradiation from a caesium source. The irradiated mice and normal, unirradiated mice were injected with 5 mg/kg per day KGF or vehicle. Thickness and proliferation in the epithelium were measured. RESULTS: KGF caused epithelial thickening of the non-keratinized layers in oral epithelium in normal mice. It increased the number of nucleated layers and influenced differentiation of post-mitotic cells in the upper layers by increasing the size and number of keratohyalin granules, and the number of desmosomes. Single and fractionated doses of radiation caused inhibition of proliferation as detected by markedly reduced BrdU incorporation following exposure, followed by epithelial atrophy. KGF treatment of mice reversed the inhibition of proliferation and atrophy that occurred in control irradiated mice. CONCLUSION: These data show that KGF reverses epithelial atrophy in mouse oral cavity caused by irradiation and suggest that KGF may be useful for the treatment of mucositis of the upper aerodigestive tract of patients treated with aggressive regimens of radiation therapy. PMID- 10374944 TI - Chromosome radiosensitivity in human G2 lymphocytes and cell-cycle progression. AB - PURPOSE: To investigate the possibility that the differential G2-phase radiosensitivity of human peripheral blood lymphocytes, found in normal individuals using the 'G2-phase chromosome radiosensitivity assay', could be attributed to heterogeneity in cellular progression to mitosis rather than differences in radiosensitivity. MATERIALS AND METHODS: Human peripheral blood lymphocytes, from four different donors, were exposed to 50 cGy X-rays and sampled at different times. The progression of cells into mitosis was monitored by 5-bromo 2'-deoxyuridine (BrdUrd) incorporation. RESULTS: The heterogeneous G2 phase chromosome radiosensitivity among different donors was abolished when homogeneous G2-phase cell populations were scored; they contained similar frequencies of cells in early or late G2-phase. CONCLUSIONS: The heterogeneous G2 phase chromosome radiosensitivity, usually found in different normal donors, is caused by the analysis of different cell populations rather than reflecting intrinsic differences in radiosensitivity. PMID- 10374945 TI - Th2 cells as effectors in postirradiation pulmonary damage preceding fibrosis in the rat. AB - PURPOSE: Radiation-induced pneumonitis and subsequent pulmonary fibrosis are important dose-limiting complications of radiotherapy. Their pathogenesis is known only in part. T-lymphocytes comprise a significant part of the infiltrating cells but little is known about their role. The aim of this study was to define the function of T-lymphocytes during development of postirradiation pneumonitis and pulmonary fibrosis. MATERIALS AND METHODS: Rats received a unilateral lung irradiation of 20 Gy. Kinetics of T-lymphocytes isolated from irradiated and non irradiated lungs were analysed. Subsequent CD4 depletion experiments were performed to affirm the importance of CD4+ T-cells in the development of lung fibrosis. Finally, the T helper-cell subtype of the T-lymphocytes was analysed by determining the cytokine mRNA by RT-PCR. RESULTS: A selective increase of CD4+ T cells was observed peaking 4 weeks after irradiation in the irradiated lungs. When rats were depleted of these cells, the postirradiation thickening of parenchyma was significantly reduced as determined by morphometric analysis of lung tissue sections. In addition, it was found that IL-4 mRNA was selectively increased in the CD4+ T-cells isolated from irradiated lungs, which indicates a lymphocyte reactivity dominated by Th2 cells. CONCLUSION: The results suggest a critical role for Th2 CD4+ T-lymphocytes in the pathogenesis of radiation-induced pneumonitis preceding lung fibrosis. PMID- 10374946 TI - Contribution of antioxidant enzymes to the adaptive response to ionizing radiation of human lymphoblasts. AB - PURPOSE: To investigate whether the adaptive response to ionizing radiation triggered by a low-dose pre-exposure could be due to the activation of the antioxidant defence system. MATERIALS AND METHODS: Human lymphoblastoid AHH-1 cells were irradiated with a 0.02 Gy gamma-radiation and 6 h later were exposed to a 3 Gy challenge dose according to a protocol allowing mutagenic adaptation. Controls included cells left unirradiated or exposed to a single dose (0.02 Gy or 3 Gy). The activities of the main cellular antioxidant enzymes (AOE) - copper zinc superoxide dismutase (SOD1), manganese superoxide dismutase (SOD2), catalase (CAT), glutathione peroxidase (GPX), glutathione-S-transferase (GST), glutathione reductase (GSR) and glucose 6-phosphate dehydrogenase (G6PD) - were evaluated at different times after treatment. The levels of SOD2 and CAT proteins were also analysed using the immuno Western blot method. RESULTS: Compared with non irradiated controls, the effect of 3 Gy alone was shown to increase GPX and CAT activities at 20 h after irradiation. Pre-exposure of cells did not change these late alterations. Soon after irradiation the activities of SOD2, GST, GPX and CAT were slightly higher in adapted than in non-adapted cells. CONCLUSION: The data suggest that the increased activities of some AOE observed soon after the challenge dose would result in a rapid scavenging of radicals and consequently less damage in adapted cells. Due to the moderate alterations of these AOE, the activation of antioxidant defences would only partly contribute to the protective mechanism underlying the radioadaptation of AHH-1 lymphoblasts. PMID- 10374947 TI - Effects of 50 Hz magnetic field exposure on the rate of RNA synthesis by normal human fibroblasts. AB - PURPOSE: To investigate whether exposure to magnetic fields can affect the rate of RNA synthesis, a broad measure of cellular activity. MATERIALS AND METHODS: Normal human fibroblasts were exposed to 50 Hz magnetic fields at a range of flux densities between 2 microT and 20 mT. The rate of synthesis of total RNA was determined by following the incorporation of [3H]uridine into macromolecular material. In addition, polyadenylated RNA was isolated and used to estimate the rate of synthesis of mRNA. RESULTS: Incorporation of [3H]uridine into both total and messenger RNA increased progressively throughout the 5 h exposure period in all cells. However, magnetic field exposure had no detectable effect on the rate of synthesis of either total or messenger RNA when compared with controls. CONCLUSIONS: These findings indicate that under the conditions examined, gross transcription rates are not affected by exposure to power frequency magnetic fields. Taken together with previous data, this suggests that if magnetic fields do alter cellular activity, the effect is likely to be extremely subtle. PMID- 10374948 TI - Translocation frequencies for low to moderate doses remain constant over time. PMID- 10374949 TI - Recombination in RNA viruses and in virus-resistant transgenic plants. PMID- 10374950 TI - The nine C-terminal residues of the grapevine fanleaf nepovirus movement protein are critical for systemic virus spread. AB - The grapevine fanleaf virus (GFLV) RNA2-encoded polyprotein P2 is proteolytically cleaved by the RNA1-encoded proteinase to yield protein 2A, 2B(MP) movement protein and 2C(CP) coat protein. To further investigate the role of the 2B(MP) and 2C(CP) proteins in virus movement, RNA2 was engineered by alternatively replacing the GFLV 2B(MP) and 2C(CP) genes with their counterparts from the closely related Arabis mosaic virus (ArMV). Transcripts of all chimeric RNA2s were able to replicate in Chenopodium quinoa protoplasts and form tubules in tobacco BY-2 protoplasts in the presence of the infectious transcript of GFLV RNA1. Virus particles were produced when the GFLV 2C(CP) gene was replaced with its ArMV counterpart, but systemic virus spread did not occur in C. quinoa plants. In addition, chimeric RNA2 containing the complete ArMV 2B(MP) gene was neither encapsidated nor infectious on plants, probably because polyprotein P2 was incompletely processed. However, chimeric RNA2 encoding ArMV 2B(MP), in which the nine C-terminal residues were those of GFLV 2B(MP), formed virus particles and were infectious in the presence of GFLV but not ArMV 2C(CP). These results suggest that the nine C-terminal residues of 2B(MP) must be of the same virus origin as the proteinase for efficient proteolytic processing of polyprotein P2 and from the same virus origin as the 2C(CP) for systemic virus spread. PMID- 10374951 TI - Effect of C-terminal deletions in the movement protein of cowpea chlorotic mottle virus on cell-to-cell and long-distance movement. AB - In order to elucidate the function of the C-terminal region of cowpea chlorotic mottle bromovirus (CCMV) movement protein (MP) in cell-to-cell movement, a set of deletions ranging from 10 to 80 amino acids (deltaMP10, deltaMP20, deltaMP33, deltaMP43, deltaMP60 and deltaMP80) was engineered into the MP gene encoded by the biologically active clone C3/deltaCP-EGFP, a variant of CCMV RNA3 that contained wild-type (wt) MP and the enhanced green fluorescent protein (EGFP) gene in place of the coat protein (CP). The effect of each MP deletion on cell-to cell movement was examined in three susceptible host plants: Chenopodium quinoa, Nicotiana benthamiana and cowpea (Vigno sinensis cv. Black Eye). The results indicate that, except for mutant deltaMP43, infections resulting from the deletion mutants remained subliminal. Interestingly, infections resulting from inoculating mutant deltaMP43, which lacked the 43 most C-terminal amino acids, spread rapidly between cells and the number of infected cells expressing EGFP approached that of control inoculations made with C3/deltaCP-EGFP. To verify whether the presence of wt CP altered the movement behaviour of these mutants, each MP deletion was also incorporated into the genetic background of wt CCMV RNA3 (pCC3) and inoculated independently to all three hosts. The results suggest that the overall movement process exhibited by each MP mutant is influenced profoundly by the presence of CP and the particular host plant tested. PMID- 10374952 TI - Persistent virus integration into the genome of its algal host, Ectocarpus siliculosus (Phaeophyceae). AB - The brown alga Ectocarpus siliculosus frequently carries an endogenous virus, E. siliculosus virus (EsV-1), the genome of which is a circular, double-stranded DNA molecule of about 320 kbp. After infection, which occurs in the unicellular spores or gametes, the virus is present latently in all somatic cells of the host. Virus multiplication is restricted to cells of the reproductive organs. It has been an open question whether the latent viral DNA occurs as a free episome or becomes integrated into the host genome. PCR studies showed that viral DNA co migrates with high molecular mass DNA in pulsed-field gel electrophoresis, which confirms that latent viral DNA is integrated into the host genome. PMID- 10374953 TI - Poly(C)-binding protein interacts with the hepatitis C virus 5' untranslated region. AB - We have investigated whether poly(C)-binding protein (PCBP)-1 and PCBP-2 interact with the hepatitis C virus (HCV) 5' untranslated region. Our results demonstrate that glutathione S-transferase (GST)-PCBP-1 and GST-PCBP-2 fusion proteins bind specifically to the HCV 5' untranslated region. An antiserum raised against PCBP 2 induced a supershift after incubation with RNA-protein complexes formed between proteins in a HeLa cell cytoplasmic extract and the HCV 5' untranslated region, indicating that this interaction occurs intracellularly. The complete internal ribosome entry site was necessary for efficient binding, suggesting that maintenance of the secondary structure was necessary for recognition of the binding site by the PCBPs. PMID- 10374954 TI - Evidence of increasing diversification of hepatitis C viruses. AB - Hepatitis C virus (HCV) has high genomic variability and, since its discovery, at least six different types and an increasing number of sub-types have been reported. The HCV genotype may influence virus replication, the natural course of disease and the response to therapy. HCV has been described as a dynamic population of heterogeneous, closely related variants, designated quasispecies. In order to study the degree of genetic variability of strains isolated in Montevideo, Uruguay, sequence data obtained from the 5' non-coding region of type 1-infected patients were compared with published sequences from 53 different strains of all types isolated all over the world. The phylogenetic analysis revealed that type 1 strains isolated in Montevideo represent a different genetic lineage from major sub-types 1a and 1b strains and indicates an increasing diversification of HCV viruses. PMID- 10374955 TI - Identification of nucleocapsid protein residues required for Sendai virus nucleocapsid formation and genome replication. AB - Alanine substitution mutations in the Sendai virus nucleocapsid (NP) protein have defined highly conserved hydrophobic and charged residues from amino acids (aa) 362 to 371 that are essential for function of the protein in RNA replication. Mutant NP362, which had the change F362A, was incapable of supporting in vitro RNA replication. NP362 expressed alone formed extended oligomers which exhibited an abnormal morphology and density suggesting that these particles were not associated with any RNA. Mutant NP364, which had changes L362A and G365A, was also inactive in RNA replication; however, this was because the protein was unstable and did not form NP-NP complexes. Mutant NP370 mutant, which had changes K370A and D371A, was inactive in in vitro replication, although it could form the required NP0-P and NP-NP protein complexes. The self-assembled nucleocapsid-like particles formed by NP370 alone had a morphology like that of wild-type NP and banded in CsCl as ribonucleoprotein particles, suggesting that they contained cellular RNA. These data suggest that the replication defect of NP370 may be in the ability to specifically encapsidate Sendai virus genome RNA. Mutant NP373, where nonconserved charged residues at aa 373 and 375 were substituted with alanine, gave a wild-type phenotype. Thus these amino acids are not required for either protein-protein interactions or in vitro Sendai virus RNA replication. PMID- 10374956 TI - Long-term CD8+ T cell memory to Sendai virus elicited by DNA vaccination. AB - The capacity of DNA vaccines to prime CD8+ T cells makes them excellent candidates for vaccines that are designed to emphasize cellular immunity. However, the long-term stability of CD8+ T cell memory induced by DNA vaccination is poorly characterized. Here, the quality of CD8+ T cell recall responses in mice was investigated more than 1 year after DNA vaccination with the Sendai virus nucleoprotein gene. Cytotoxic T lymphocyte (CTL) activity specific for both dominant and subdominant epitopes could be recalled readily 1 year after vaccination and the frequencies of CTL precursors specific for both of these epitopes were relatively high. These CTL responded strongly to subsequent Sendai virus infection in terms of their ability to migrate to the lung and to differentiate into effector cells. In addition, the recall response to virus infection, as determined by CTL activity in the lungs and IFN-gamma responses in the spleen, was both faster and greater in magnitude than that in control immunized mice. Significantly, virus titres were reduced at least 100-fold in the lungs of mice that were immunized more than 1 year before infection, as compared with control mice. These data demonstrate that CD8+ T cell memory elicited by DNA vaccination is functionally relevant and persists for at least 1 year. PMID- 10374957 TI - Synergistic effect of immunization with a peptide cocktail inducing antibody, helper and cytotoxic T-cell responses on protection against respiratory syncytial virus. AB - Respiratory syncytial virus (RSV)-specific cytotoxic T lymphocytes (CTL) or neutralizing antibodies can protect against RSV infection when induced separately by immunization with synthetic peptides. In the work described here, RSV-specific neutralizing antibodies and CTLs were induced after immunization with a cocktail of peptides consisting of a B-cell mimotope (S1S-MAP), a T-helper epitope (SH:45 60) and a CTL epitope linked to a fusion (F) peptide (F/M2:81-95) that were comparable to those induced by the peptides alone. Following challenge, a 190 fold reduction in RSV titre was observed in the lungs of peptide cocktail immunized mice. The combination of RSV-specific humoral and cellular immunity induced by the peptide cocktail was thus more effective at clearing RSV than peptide-induced humoral or cellular immunity alone. PMID- 10374958 TI - Sequence analysis of VP1 and VP7 genes suggests occurrence of a reassortant of G2 rotavirus responsible for an epidemic of gastroenteritis. AB - G2 rotavirus was prevalent in a 1993 epidemic of acute gastroenteritis in Taiwan. In this study, the genetic relationship among G2 rotavirus strains was analysed. The VP7 genes were amplified and sequenced. Except for one strain isolated in 1981, the nucleotide sequences of the VP7 genes of most of the G2 rotaviruses were very similar (identity > 97%) and were closely related to that of a Japanese G2 reference strain, S2. The genetic relatedness of G2 rotaviruses was analysed further by RNA-RNA hybridization. The genomes of the major G2 strains of 1993 did not hybridize well with those of the G2 strains of previous seasons in RNA segments 1, 6 and 7. Partial nucleotide sequences of the VP1 gene were analysed and appeared to be similar among the major G2 strains from the same epidemic (identity > 98%), whereas the identity of the VP1 genes of the major G2 strains of the 1993 epidemic to those of previous seasons was only about 84%. Since the numbers of mutations accumulated in the VP1 and VP7 genes over a period of 10 years were comparable, the significant change in the VP1 genes of the major strains of the 1993 epidemic suggests that these G2 rotaviruses had evolved by genetic reassortment. PMID- 10374959 TI - Serum and intestinal isotype antibody responses to Wa human rotavirus in gnotobiotic pigs are modulated by maternal antibodies. AB - The effects of passive antibodies on protection and active immune responses to human rotavirus were studied in gnotobiotic pigs. Pigs were injected at birth with saline or sow serum of high (immunized) or low (control) antibody titre and subsets of pigs were fed colostrum and milk from immunized or control sows. Pigs were inoculated at 3-5 days of age and challenged at 21 days post-inoculation (p.i.) with virulent Wa human rotavirus. Pigs receiving immune serum with or without immune colostrum/milk were partially protected against diarrhoea and virus shedding after inoculation, but had significantly lower IgA antibody titres in serum and small intestinal contents at 21 days p.i. and lower protection rates after challenge compared with pigs given control or no maternal antibodies. IgG antibody titres were consistently higher in small than in large intestinal contents. Pigs given control serum with control colostrum/milk had lower rates of virus shedding after inoculation than those given control serum alone. In summary, high titres of circulating maternal antibodies with or without local (milk) antibodies provided passive protection after inoculation but suppressed active mucosal antibody responses. These findings may have implications for the use of live, oral rotavirus vaccines in breast-fed infants. PMID- 10374960 TI - Human T-cell leukaemia/lymphoma virus type 1 syncytium formation is regulated in a cell-specific manner by ICAM-1, ICAM-3 and VCAM-1 and can be inhibited by antibodies to integrin beta2 or beta7. AB - Human T-cell leukaemia/lymphoma virus type 1 (HTLV-1) is a pathogenic retrovirus responsible for a number of inflammatory pathologies and adult T-cell leukaemia. Although T-cell tropic in vivo, HTLV-1 can infect a wide variety of cell types in vitro. Cell-to-cell spread of HTLV-1 may require specific binding of envelope to its cellular receptor, with other cell-surface molecules facilitating fusion. Here it is shown that intercellular adhesion molecule-1 or -3 (ICAM-1, ICAM-3) or vascular cell adhesion molecule-1 (VCAM-1) are required for syncytium formation of K562 with HTLV-1-infected MT2 cells but not C91-PL cells. The effect of ICAMs and VCAM-1 on MT2-induced fusion can be blocked by antibodies that bind beta integrins. These fusion co-factor molecules are effective only when present in combination with HTLV-1 receptor-bearing cells and are not sufficient to mediate syncytium formation alone. The results suggest that engagement of HTLV-1-infected cells with susceptible target cells requires the simultaneous binding of viral envelope glycoprotein to the cellular receptor and co-factor molecules to beta integrins. The tissue-specific expression of adhesion molecules might therefore influence HTLV-1 virus tropism and pathogenic changes associated with syncytium formation. PMID- 10374961 TI - Ovine lentivirus-infected macrophages mediate productive infection in cell types that are not susceptible to infection with cell-free virus. AB - Ovine lentiviruses and caprine arthritis-encephalitis virus (CAEV) are prototypic lentiviruses that replicate predominantly in macrophages of infected animals. In situ hybridization of pathologically affected tissues from diseased animals has shown that viral RNA exists in permissive macrophages as well as in non macrophage cell types that do not support productive virus replication. These findings raise questions about the cellular tropism of these viruses in vivo and how this may relate to their pathogenesis and the establishment of persistent infections. In this study, the susceptibility of macrophages and fibro-epithelial cells derived from goat synovial membrane (GSM) to infection by 14 North American ovine lentivirus strains was examined. All 14 strains were macrophage-tropic, as indicated by expression of viral proteins and by fusion and development of syncytial cytopathic effects following co-culture of infected macrophages with GSM cells. In contrast, neither viral DNA nor viral proteins was detected in GSM cells inoculated with cell-free virus from nine of the 14 strains. Specific virus proteins were immunoprecipitated from restrictive GSM cells following culture with infected macrophages and serial passage of GSM cells to remove the macrophages. The lack of infection of GSM cells by cell-free virus from some ovine lentivirus field strains was circumvented by cell-associated virus infection from infected macrophages to GSM cells following cell-to-cell contact. This strategy could be one of the mechanisms involved in the escape from immune surveillance and establishment of persistent infection in infected animals. PMID- 10374962 TI - An active foamy virus integrase is required for virus replication. AB - Foamy viruses (FVs) make use of a replication strategy which is unique among retroviruses and shows analogies to hepadnaviruses. The presence of an integrase (IN) and obligate provirus integration distinguish retroviruses from hepadnaviruses. To clarify whether a functional IN is required for FV replication, a mutant in the highly conserved DD35E motif of the active centre was analysed. This mutant was found to be able to express Gag and Pol protein precursors and cleavage products and to generate and deliver cDNA. However, this mutant was replication-deficient. The junctions of individual foamy proviruses with cellular DNA were sequenced. The findings suggest that FV integration is asymmetrical, because the proviruses started with what is believed to be the U3 end of the free linear DNA to generate the conventional TG dinucleotide, while apparently two nucleotides from the U5 end were cleaved to create the complementary CA dinucleotide. Alignment of known FV genome sequences indicated that this mechanism of integration is not restricted to the two FV isolates from which integrates were studied, but appears to be a common feature of this retrovirus subfamily. In conclusion, with respect to the necessity of a functionally active IN for virus replication FVs behave like other retroviruses; their mechanism of integration, however, is probably unique. PMID- 10374963 TI - African swine fever virus: a B cell-mitogenic virus in vivo and in vitro. AB - The two major characteristics of pathogenesis in African swine fever virus (ASFV) infections of domestic pigs are massive B-cell apoptosis and haemorrhage. The effects of ASFV on porcine B cells have therefore been systematically examined in vivo, by using virus-infected pigs and SCID-Beige mice reconstituted with porcine bone marrow, and in vitro, by using porcine B-cell lines and B cells from normal and ASFV-infected pigs. Secretion of porcine Ig was stimulated by ASFV both in vivo and in bone marrow cultures in vitro, with the virulent Malawi isolate of ASFV being the most effective. Stimulation of Ig secretion in vitro depended on the presence of ASFV-infected macrophages and did not occur with supernatants from ASFV-infected macrophages. Although the virus alone did not stimulate proliferation of purified B cells in vitro, it was co-stimulatory with CD154 (CD40 ligand). The B cells recovered from ASFV-infected porcine lymphoid tissue were of activated surface marker phenotypes and, interestingly, expressed diminished levels of the B-cell co-stimulatory surface molecule CD21. In addition, they were highly sensitive to IL-4 and CD154. These results may be integrated into a model of pathogenesis in which those B cells activated indirectly as a result of virulent ASFV infection of macrophages are not rescued from apoptosis through interaction with CD154, due to the drastic depletion of T cells that occurs early in infection. The consequently diminished specific anti ASFV antibody response would favour survival of the virus, with the non-specific hypergammaglobulinaemia being perhaps another example of pathogen-mediated immune deviation. PMID- 10374964 TI - Vaccinia virus-bacteriophage T7 expression vector for complementation analysis of late gene processes. AB - A vaccinia virus-bacteriophage T7 RNA polymerase hybrid transient expression vector has been developed for complementation analysis of late gene functions in vaccinia virus. The conditionally defective virus ts21 was modified to express the bacteriophage T7 RNA polymerase. The derived virus, vtsT7, was conditionally defective in viral late gene expression but produced high levels of a target protein under the control of a T7 promoter at non-permissive temperatures. The level of beta-galactosidase expression under the control of a T7 promoter was slightly lower in vtsT7 infections than those with the prototypical T7 RNA polymerase vector vTF7.3. However, the levels of expression for the human immunodeficiency virus envelope gene, a protein which undergoes post translational modification, was slightly higher in vtsT7 infections, suggesting that some proteins may be expressed better in the absence of vaccinia virus late gene expression. Infections using vtsT7 at a low m.o.i. at 39 degrees C resulted in the accumulation of high molecular mass, non-linear replicative intermediates of vaccinia virus DNA replication and high levels of expression of a transfected gene proximal to a T7 promoter. The virus vtsT7 provides a means for the analysis of potential trans-acting factors participating in vaccinia virus late processes such as resolution of DNA replicative intermediates. PMID- 10374965 TI - The unique N terminus of herpes simplex virus type 1 ribonucleotide reductase large subunit is phosphorylated by casein kinase 2, which may have a homologue in Escherichia coli. AB - Studies were performed to determine if the unique N-terminal domain of the R1 subunit from herpes simplex virus (HSV) type 1 ribonucleotide reductase is a substrate for casein kinase 2 (CK2). Transphosphorylation assays demonstrated that R1 was highly phosphorylated by this enzyme with multiple phosphorylation sites mapped to the N terminus between residues 1 and 245. Immunoprecipitation pull-down assays using R1-specific antisera failed to demonstrate a stable interaction between R1 and CK2 but residual amounts of CK2 present after immunoprecipitation efficiently transphosphorylated R1. Activity assays with a peptide substrate identified CK2 in R1 immunoprecipitated from infected-cell extracts but did not detect activity in R1 proteins immunoprecipitated from bacterial extracts. However, Western blotting identified potential E. coli homologues of the CK2 alpha and beta subunits. These results support conclusions that the N-terminal domain of HSV R1 is not a protein kinase and that all previous results can be explained by contaminating kinases, principally CK2. PMID- 10374966 TI - Epitopes on glycoprotein C of bovine herpesvirus-1 (BHV-1) that allow differentiation between BHV-1.1 and BHV-1.2 strains. AB - In cattle, bovine herpesvirus-1 (BHV-1) can cause a mild genital disease known as infectious pustular vulvovaginitis (IPV) and a more severe respiratory disease known as infectious bovine rhinotracheitis (IBR). On the basis of epidemiological data, it has been proposed that these diseases are caused by strains with different genotypes (IBR by BHV-1.1 and IPV by BHV-1.2 strains). By using a panel of 237 BHV-1 isolates, a monoclonal antibody (MAb 71) was found that failed to react with all 54 putative IPV strains in the panel, and another MAb (77) was found that did not react with 16 of these 54 IPV strains. Because MAbs 71 and 77 also failed to react with a BHV-1.1 glycoprotein C (gC)-deletion mutant, it was hypothesized that both MAbs recognize BHV-1.1 gC. By marker-rescue experiments and by expressing fragments of the BHV-1.1 gC gene in recombinant baculoviruses, it was shown that both MAbs indeed recognize BHV-1.1 gC. MAb 71 recognizes the N terminal half and MAb 77 recognizes the C-terminal half of BHV-1.1 gC. In a PEPSCAN analysis with 12-mer oligopeptides, MAb 71 reacted with overlapping peptides containing gC amino acid residues 75-80 and MAb 77 did not react in this analysis. The differences in gC found in this study may contribute to the biological differences between BHV-1.1 and BHV-1.2. PMID- 10374967 TI - N- and C-terminal external domains of human herpesvirus-6 glycoprotein H affect a fusion-associated conformation mediated by glycoprotein L binding the N terminus. AB - Human herpesvirus-6 (HHV-6), like other betaherpesviruses, shows cell fusion with wild-type strains, and this cellular spread is mediated by the glycoprotein gH/gL complex. Anti-fusion monoclonal antibodies (MAbs) specific for HHV-6 glycoprotein gH inhibit infection and prevent cellular spread by syncytia formation. Reactivity of these MAbs with gH deletion mutants suggests a conserved C-terminal fusion-associated domain. A conserved motif here has an N-glycosylation site and characteristics of a beta turn. Motif deletion abrogated MAb recognition while co expression with glycoprotein gL restored this conformational epitope, indicating the importance of folding and not glycosylation at this site. Our previous studies showed gL binding to gH at an N-terminal domain specific for betaherpesviruses. To further examine the function of this N-terminal domain, a betaherpesvirus-specific motif was deleted. This mutant gH still bound gL, and was recognized by the anti-fusion MAbs; however, recognition was now primarily in the immature form and reduced during processing to the mature form. A model is discussed whereby gL binding gH at the N-terminal domain acts to draw together the C-terminal extracellular domain and this interaction affects a functional conformation during glycoprotein maturation. PMID- 10374968 TI - Variation within the glycoprotein B gene of human cytomegalovirus is due to homologous recombination. AB - Human cytomegalovirus (HCMV) strains can be classified into different glycoprotein B (gB) genotypes. In a previous study, frequent intragenic variation of the gB gene was shown. The aim of this study was to analyse whether gB variation was due to homologous recombination. The gB gene of DNA extracts derived from the peripheral blood leukocytes of 14 immunosuppressed patients was amplified by PCR and cloned. Three variable sites of gB were analysed by restriction fragment analysis and DNA sequencing and compared with published prototypic strains. In three patients doubly infected with two distinct HCMV gB strains, prototypic (60-85%) and non-prototypic recombinant strains (5-40%) were detected. To demonstrate that homologous recombination is driving HCMV gB variability, cells were coinfected with plaque-purified prototypic gB strains and recombinant gB genes were selectively amplified by PCR. gB recombinants were detected after 15 days of coculture and cross-over sites were determined by sequencing. These data indicate that homologous recombination contributes to the variability of the gB gene in vitro and in vivo. PMID- 10374969 TI - Definition of the transcription factors which bind the differentiation responsive element of the Epstein-Barr virus BZLF1 Z promoter in human epithelial cells. AB - Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus and an important human pathogen. Initiation of the EBV lytic cycle is dependent upon transcription of the EBV BZLF1 gene. Our previous studies of transcriptional regulation of the BZLF1 Z promoter (Zp) in human SCC12F epithelial cells identified a region within Zp that is responsive to epithelial cell differentiation. In the present study, we localize this differentiation responsive element to the CREB/AP-1-like binding site (TGACATCA) between -67 to -60 bp within Zp, previously designated ZII, and furthermore show that homodimers and heterodimers of CREB and ATF-1 specifically bind ZII. Consistent with a regulatory role for CREB and ATF-1 in differentiation dependent BZLF1 expression, ZII was able to bind approximately 3-fold more CREB and ATF-1 when incubated with nuclear extract obtained from populations of SCC12F cells enriched for the differentiated phenotype than when incubated with extract obtained from populations enriched for the undifferentiated phenotype. In addition, CREB and ATF-1 were found to increase in abundance during SCC12F differentiation. These results indicate a regulatory role for CREB and ATF-1 in differentiation-dependent expression of BZLF1 in human epithelial cells. PMID- 10374970 TI - Human papillomavirus (HPV) E6 interactions with Bak are conserved amongst E6 proteins from high and low risk HPV types. AB - Human papillomavirus (HPV) replication occurs in terminally differentiating epithelium, and requires the activation of cellular DNA replication proteins. Unscheduled DNA replication can result in the induction of apoptosis, and the viral E6 protein induces the degradation of p53 to prevent this. It has recently been shown that HPV-18 E6 can also stimulate the degradation of Bak, a pro apoptotic member of the Bcl-2 family. This report shows that the E6 proteins from HPV-18, HPV-16 and HPV-11 can all bind to Bak in vitro, stimulate its degradation in vivo and reduce Bak-induced apoptosis. However, the non-oncogenic HPV-11 E6 is less effective than the oncogenic E6 proteins in each of these assays, indicating that the ability of HPV to circumvent the apoptosis induced by Bak may contribute to the oncogenic potential of the virus. PMID- 10374971 TI - Molecular determinants of adenovirus serotype 5 fibre binding to its cellular receptor CAR. AB - Adenovirus (Ad) tropism is mediated in part through the fibre protein. The common coxsackie B virus and Ad receptor (CAR) was recently identified as the major receptor for subgroup C Ad serotype 5 (Ad5) and serotype 2 (Ad2) fibres. Effects of mutations in the Ad5 fibre gene were studied to assess domains of the fibre capsomer that could alter virus tropism without altering virus assembly and replication. All mutants that accumulated as fibre monomers failed to assemble with a penton base and proved lethal for Ad5 which suggests that the absence of infectious virions resulted in part from a defect in fibre penton base assembly. Cell binding capacity of all fibre mutants was investigated in cell binding competition experiments with adenovirions using CHO-CAR cells (CHO cells that have been transfected with CAR cDNA and express functional CAR). The results suggest that the R-sheet of the Ad5 fibre knob monomer contains binding motifs for CAR and that beta-strands E and F, or a region close to them, may also be involved in receptor recognition. PMID- 10374972 TI - In vivo activity of a mixture of two human monoclonal antibodies (anti-HBs) in a chronic hepatitis B virus carrier chimpanzee. AB - A 35-year-old female hepatitis B virus carrier chimpanzee was infused with one dose of a mixture of human monoclonal antibodies 9H9 and 4-7B (antibodies against hepatitis B virus surface antigen; HBsAg). Blood samples were taken before and up to 3 weeks after infusion. HBsAg and antibodies against HBsAg (anti-HBs) were quantified by radioimmunoassay and enzyme immunoassay. Free anti-HBs was never detected. Thirty min after the start of the infusion the HBsAg level was minimal with maximum loading of the chimpanzee HBsAg with human immunoglobulin. HBsAg complexes could be dissociated by acid treatment. The HBsAg level was completely restored on day 7. Similar results were obtained for the preS1-containing particles that may represent the infectious viral particles in the chimpanzee serum. A mouse monoclonal anti-HBs (HBs.OT40) was found to compete with 9H9 in artificial immune complexes with the pre-treatment HBsAg from the chimpanzee. Used as a conjugate, HBs.OT40 yielded a maximum decrease in the signal in the 30 min sample compared to non-competing anti-HBs conjugates. This indicates binding of HBsAg with 9H9 in the circulation of the chimpanzee. Immune-complexed 4-7B could not be detected by its corresponding 4-7B peptide conjugate, probably due to its low concentration in the complexes. It is concluded that human monoclonal anti-HBs can effectively reduce the level of HBsAg in serum from this chronic carrier. Monoclonals 9H9 and 4-7B may complement each other due to their different mechanisms of inactivation, probably with higher efficiency than that monitored by our HBsAg screening assays. PMID- 10374973 TI - Circular configuration of the genome of ascoviruses. AB - A circular configuration of genomic DNA was observed in ascoviruses isolated from two species of insects of the family Noctuidae [fall armyworm (Spodoptera frugiperda) and cotton bollworm (Helicoverpa zea)] using restriction endonuclease (REN) digestion, conventional gel electrophoresis, pulsed-field gel electrophoresis and Southern hybridization analysis. This circular configuration of ascovirus genomic DNA was established based on the difference between linear and circular DNA in the numbers of fragments resolved on agarose gel electrophoresis after single and double REN digestion. Genomic DNA of ascoviruses was found to be sheared after purification. PMID- 10374974 TI - The nucleotide sequence of sacbrood virus of the honey bee: an insect picorna like virus. AB - We have determined the nucleotide sequence of sacbrood virus (SBV), which causes a fatal infection of honey bee larvae. The genomic RNA of SBV is longer than that of typical mammalian picornaviruses (8832 nucleotides) and contains a single, large open reading frame (179-8752) encoding a polyprotein of 2858 amino acids. Sequence comparison with other virus polyproteins revealed regions of similarity to characterized helicase, protease and RNA-dependent RNA polymerase domains; structural genes were located at the 5' terminus with non-structural genes at the 3' end. Picornavirus-like agents of insects have two distinct genomic organizations; some resemble mammalian picornaviruses with structural genes at the 5' end and non-structural genes at the 3' end, and others resemble caliciviruses in which this order is reversed; SBV thus belongs to the former type. Sequence comparison suggested that SBV is distantly related to infectious flacherie virus (IFV) of the silk worm, which possesses an RNA of similar size and gene order. PMID- 10374975 TI - Brain injury does not modify transmissible spongiform encephalopathy caused by intraperitoneal inoculation with Fukuoka-1 strain. AB - The pathogenesis of neuroinvasion in Creutzfeldt-Jakob disease and other transmissible spongiform encephalopathies following the peripheral uptake of a disease agent is still not fully understood. The possibility of neuroinvasion either being established or being accelerated by an insult to the brain has not previously been tested. The experiment described herein was designed to examine this possibility by wounding the brain following an intraperitoneal challenge with a mouse-adapted strain of human transmissible spongiform encephalopathy, Fukuoka-1 strain. The results showed that brain injury introduced in any period before the appearance of cerebral abnormal prion protein deposition modified neither the clinical features, including the incubation period, nor the histopathology of the mice. Our findings suggest that neurovirulence of the agent may not be sufficiently promoted by brain injury. PMID- 10374976 TI - An in vivo evaluation of the reproducibility of intima-media thickness measurements of the carotid artery segments using B-mode ultrasound. AB - B-mode ultrasound may be used to measure the intima-media thickness (IMT) in subjects with a history of atherosclerosis. The variability between measurements depends on the subjective interpretation of ultrasonographers and readers. The two carotid arteries, subdivided in common (CCA), bulbus (BUL) and internal (ICA) of 10 men with proven coronary disease, were scanned twice by two ultrasonographers with a 1-week interval. The IMTs were measured off-line by two readers. The number of IMT measurements was 75 (94%) of 80 in the CCA, 61 (76%) of 80 in the BUL and 43 (54%) of 80 in the ICA segment. In the CCA segment, the agreement between readers (mean = 0.02 mm; limits: -0.26 to +0.3 mm) and between visits for each reader separately (reader 1: mean = 0.01 mm; limits: -0.33 to +0.35 mm and, reader 2: mean = 0.04 mm; limits: -0.36 to +0.44 mm) was better than in the more distal segments. Therefore, it is concluded that IMT measurements are best performed in the CCA segment. PMID- 10374977 TI - Effect of acquisition rate on liver and portal vein enhancement with microbubble contrast. AB - We showed that tissue enhancement with microbubbles is dependent upon transmit power. Because intermittent imaging decreases bubble exposure to ultrasound, and also decreases the ability of the sonographer to maintain anatomic orientation, we aimed to determine the optimum frame rate that maximizes enhancement and allows for continued anatomic orientation. Seven rabbits with an avascular liver lesion created by percutaneous injection of 1 mL ethyl alcohol 7 days earlier were imaged with an Acuson 128XP/10 using a 7-MHz sector transducer at fixed transmit power. Each rabbit was imaged 5 times in random order, at 1 frame/30 s, 1frame/5 s, 1frame/s, 4 frames/s, and 28 frames/s. The same plane was imaged at all frame rates from before to 15 min after the bolus injection of 0.3-mL (0.1 0.12 mL/kg) of AF0150 (Imagent, Alliance Pharmaceutical Corp., San Diego, CA). Liver and portal vein videointensity relative to the lesion were evaluated over time. In this study, liver enhancement progressively increased as the frame rate was reduced (p<0.001). Peak, duration, and area under the time-intensity curve were all greater at the lower frame rates (1 fr/30 s, 1 fr/5 s, and 1 fr/s) than at 28 fr/s (p<0.05). Anatomic orientation was maintained at 1 frame/s rate at which peak enhancement was 44% greater and duration was 100% longer than at 28 frames/s (p<.05). Portal vein enhancement was not affected by frame rate. In conclusion, with intermittent imaging, enhancement was dependent upon frame rate and the ability of the region being imaged to replenish its bubbles between consecutive acquisitions. The 1 frame/s allowed for anatomic orientation and adequate tissue contrast. PMID- 10374979 TI - Spectrogram analysis of arterial Doppler signals for off-line automated hits detection. AB - Recently, a time processing of arterial Doppler signals was proposed to detect automatically high-intensity transient signals (HITS). This technique provided satisfactory detection results, but was not always constantly accurate, particularly with high-resistance blood velocity profiles. A time-frequency processing, based on the spectrogram, is presented to detect the presence of emboli in the arterial Doppler signals. The method uses the narrow-band hypothesis and extracts the detection criterion from the time-frequency representation (TFR). A first database of 560 peripheral arterial Doppler HITS was created to study microemboli and to define the normal limits to be used in our method. A threshold was experimentally defined using this database, and then applied to 38 recordings from 12 patients. Using another database, 6 human expert Doppler users reported 140, 176, 155, 161, 161 and 146 HITS, corresponding to a total of 197 different observed HITS. When an event was detected by at least 6, 5, 4, 3, 2 and 1 of the observers, sensitivity of the automatic detection was 93.9, 91.7, 89.6, 88.7, 84.7 and 73.1%, respectively. The sensitivity of our automatic detection is, thus, highly associated with the number of observers in agreement. A preliminary experiment has been performed to test the method in the case of long recording duration. In 15 patients, 6 h 24 min of recordings have been analyzed. The proposed automated processing provided an overall sensibility of 91.5%. The present work shows that this detection scheme preserves good sensibility and improves the positive predictive value compared with the time processing recently proposed. PMID- 10374978 TI - Semiautomatic registration of volumetric ultrasound scans. AB - We demonstrate the ability to register easily and accurately volumetric ultrasound scans without significant data preprocessing or user intervention. Two volumetric ultrasound breast scan data sets were acquired from two different patients with breast cancer. Volumetric scan data were acquired by manually sweeping a linear array transducer mounted on a linear slider with a position encoder. The volumetric data set pairs consisted of color flow and/or power mode Doppler data sets acquired serially on the same patients. A previously described semiautomatic registration method based on maximizing mutual information was used to determine the transform between data sets. The results suggest that, even for the deformable breast, three-dimensional full affine transforms can be sufficient to obtain clinically useful registrations; warping may be necessary for increased registration accuracy. In conclusion, mutual information-based automatic registration as implemented on modern workstations is capable of yielding clinically useful registrations in times <35 min. PMID- 10374980 TI - Umbilical arterial blood flow in the mouse embryo during development and following acutely increased heart rate. AB - In anesthetized, pregnant ICR mice, we measured embryonic umbilical arterial velocity at baseline and during bipolar atrial or ventricular pacing. Pregnant mice were anesthetized with pentobarbital (60 mg/kg intraperitoneal) and ventilation was mechanically supported via a tracheotomy. Embryos were exposed through a mid-line laparotomy and regional hysterotomy. We recorded umbilical velocity using a 1-mm diameter piezoelectric crystal and 20-MHz, pulsed Doppler velocimeter at embryo day (ED) 10.5 (n = 8), 12.5 (n = 10), 13.5 (n = 27), 14.5 (n = 12), and 16.5 (n = 17). We then acutely altered embryonic heart rate in n = 8 ED 13.5 mouse embryos by bipolar atrial and ventricular pacing. Embryonic heart rate in this experimental preparation increased from 123+/-7 to 193+/-11 beats/min from ED 10.5 to 16.5 (p<0.05). Peak instantaneous average velocity increased from 21+/-2 to 55+/-6 mm/s from ED 10.5 to 16.6 (p<0.05), as did stroke volume and blood flow (p<0.05 for each). In contrast to human umbilical arterial velocity profiles, significant forward diastolic flow was not seen at these stages, suggesting higher placental resistance in mice versus humans at comparable developmental time points. As previously noted for the chick embryo, murine embryonic umbilical arterial velocity decreased after atrial pacing and disappeared after ventricular pacing. Thus, we can determine embryonic umbilical blood flow during the overlapping periods of murine cardiac and placental morphogenesis. PMID- 10374981 TI - Noninvasive cerebrospinal fluid shunt flow measurement by Doppler ultrasound using ultrasonically excited bubbles: a feasibility study. AB - Because normal cerebrospinal fluid (CSF) has almost no natural Doppler scatterers, patency testing of ventriculoperitoneal cerebrospinal fluid shunts (small silastic tubing with lumen diameter of approximately 1 mm draining excessive CSF from the brain) cannot be performed by Doppler ultrasound. We have developed a low-frequency bubble excitation system that generates microbubble scatterers in both distilled water and CSF. Doppler ultrasound can then be used for flow measurement in a ventriculoperitoneal shunt. By using low duty-cycle (approximately 10%), low-frequency (approximately 30 kHz), and low-amplitude (approximately 30 kPa) ultrasound, a population of microbubbles can be maintained for sufficiently long times (>10 min) for Doppler ultrasound measurement, although bubble initiation is inconsistent. The minimum pressure needed for bubble maintenance was found to decrease with increasing burst length and duty cycle. It has been possible to detect the presence of CSF shunt flow down to a mean flow rate of 3 mL/h (mean velocity approximately 0.6 mm/s). The bubble maintenance scheme developed satisfies the safety parameters specified by the American Institute of Ultrasound in Medicine (AIUM) and the US Food and Drug Administration (FDA). Results from both in vitro and in vivo (externalized shunts) experiments indicate the feasibility of this scheme for determining realistic CSF shunt flows, though some practical problems remain before the technique will be ready for clinical use. PMID- 10374982 TI - Finite beam-width ray model for geometric spectral broadening. AB - The purpose of the study was to compare measured spectral width and maximum frequency with that predicted from ray models of geometric spectral broadening. Zero and finite beam-width models were used. Spectral data were acquired from a string phantom using two commonly-used linear array systems. Beam width and Doppler aperture sizes were measured using a needle hydrophone. The results showed that the experimentally measured data agreed best with the finite beam width model. The zero beam-width model was in error by up to 50% for calculated spectral width, and up to 10% for maximum frequency. It is concluded that spectral width and maximum frequency are best calculated using the finite beam width model, and that ultrasound manufacturers could improve the variation in spectral broadening measured at different locations on a single machine by adjusting the aperture size to give a constant subtended angle and beam width. PMID- 10374983 TI - Echo decorrelation estimated from signal powers. AB - Volume flow can be estimated from the decorrelation of radiofrequency (RF) intravascular ultrasound signals. The method is based on a rather time-consuming process that measures the decorrelation slope from a time signal sequence. To improve the speed of flow processing, a more efficient way of estimating the flow velocity from the ratio between the power of the temporal averaged signal and the mean signal power is described in this paper. The relationship between the signal power-ratio index and the decorrelation slope was analyzed and tested using computer-simulated data. Volumetric flow data obtained with the power-ratio method were compared to those derived from the decorrelation slope in five patients. Results of the comparison studies indicate that no significant differences in flow measurements were found between the two methods, but the power-ratio method is able to improve the processing speed significantly. PMID- 10374984 TI - Analysis of flash echo from contrast agent for designing optimal ultrasound diagnostic systems. AB - Microbubble-based contrast agents can enhance echoes in areas of low blood flow, but the bubbles are extremely sensitive and collapse easily when exposed to ultrasound (US) irradiation. An experimental study of bubble collapse was carried out to design new functions for US diagnostic systems to detect echoes from microbubbles more efficiently. For contrast agent (Levovist) solution, a high intensity, but momentary, echo (flash echo), was observed in the first frame image after a several-second suspension of transmission, but was not seen in the second frame image. These "flash echo" signals were analyzed and categorized based on microscopic observation, and the results showed that the longevity of the microbubbles was reduced by conditions such as B-mode imaging. Next, a numerical simulation of the bubbles in liquid was performed under the same conditions as in the in vitro experiment. The results showed that even bubbles less than 1 microm in diameter expand and collapse within one pulse drive, which would generate flash echoes. The flash echo imaging system described here permits flexible intermittent scanning with variable intervals, with a variable number of frames at the trigger, and with simultaneous monitoring at low power output. Animal experiments were also conducted to evaluate the system. As the interval between frames was increased, the flash echoes gradually increased, and perfusion in the parenchyma was clearly observed with an interval of 4 s. PMID- 10374985 TI - High-frequency estimation of the ultrasonic attenuation coefficient slope obtained in human skin: simulation and in vivo results. AB - In vivo ultrasonic characterization of the skin was performed at 40 MHz by estimating the slope of the attenuation coefficient in the human dermis. The centroid algorithm was first tested on simulated backscattered RF lines with a second-order autoregressive model to carry out the spectral analysis. A relative error of less than 8.5% and a relative precision of less than 6% were predicted for a 2-mm tissue thickness and for temporal window sizes ranging from 0.25 to 0.45 micros. In vivo measurements performed on 138 healthy volunteers yielded values of the attenuation coefficient slope ranging from 0.8 to 3.6 dB/cm MHz. A decrease was observed with advancing age, but no significant difference appeared between men and women. The results from this study suggest that this acoustic parameter shows the effect of the ageing process on normal skin tissue in vivo. PMID- 10374986 TI - Rationale and derivation of MI and TI--a review. AB - This review article provides a summary rationale and derivation for the mechanical index (MI) and thermal index (TI) formulas presented in the body of the "Standard for real-time display of thermal and mechanical acoustic output indices on diagnostic ultrasound equipment" (AIUM/NEMA 1998). For purposes of simplicity, this standard is referred to in this article by its colloquial "nickname," the "output display standard" or ODS. This review expands on the summary information provided in Appendix A of the ODS. Numerous references are made to the root publications from which the formulas were derived. As will be discussed in the derivation notes that follow, key parts of the MI and TI models rely heavily on experimental data. This document does not attempt to do any more than describe the relevant results of the experiments. A general overview is provided in the Introduction and Rationale for the nontechnical reader and the more rigorous Models and Derivation Notes provides a detailed mathematical review of the MI and TI models. A complete set of defined terms is provided in Appendix 1. PMID- 10374987 TI - Ultrasonic enhancement of coated particle agglutination immunoassays: influence of particle density and compressibility. AB - The detection rate and sensitivity (analyte concentration limit) of coated particle agglutination immunoassays are increased in ultrasonic standing waves. The influence of particle volume, density and compressibility, properties that modify the ultrasonic radiation, and interaction forces the particles experience, on assay sensitivity with latex and silica particles in the range 0.25-1.0 microm is examined here. Streptavidin-coated 0.3-microm silica particles and 0.25-microm and 1.0-microm latex particles were examined for agglutination with biotinylated bovine serum albumin (bBSA) following exposure on axis in a 4.6-MHz radial standing wave. The lowest detection limit, 2 ng/mL bBSA, was achieved with the 0.3-microm silica. The detection limit decreased with increasing latex particle size. The limit of an ultrasound-enhanced agglutination immunoassay of rabbit antimouse immunoglobulin was 6-fold better with 1.0-microm coated silica than with equal-sized latex particles. Calculations show that the particle density dependent ultrasonic interaction force dominates the particle compressibility force for the present case. The density-dependent force on silica, but not on latex particles, is shown to be comparable in magnitude to both the long-range van der Waal's attractive force and the electrostatic repulsion between the particles. This density-dependent force may explain the improved enhancement of analyte detection by coated silica compared with latex particles. PMID- 10374988 TI - Characterization of cavitational activity in lithotripsy fields using a robust electromagnetic probe. AB - A robust electromagnetic probe has been used to investigate cavitational activity in vitro in the fields of two extracorporeal lithotripters and one intracorporeal lithotripter. Some aspects of the electromagnetic probe design and characteristics are described. A series of experiments have been carried out with results indicating that the probe head moves in response to the pressure gradient generated by radial motion of cavitation bubbles. Empirical expressions have been derived for the cavitational force acting on the probe head, and for the low frequency sawtooth pressure wave generated by inertial cavitation. This is the first time that the existence of a low-frequency sawtooth wave produced by inertial cavitation has been described. A linear relationship exists between the negative pressure amplitude of the sawtooth wave and the lifetime of the bubbles. Close to the cavitation site, substantial negative pressure is maintained throughout bubble expansion. This can easily exceed the tensile strength of urinary calculi, and may be considered to be an important mechanism for disintegrating these relatively weak structures. A pilot study has also been carried out involving three patients treated by extracorporeal lithotripsy. Signals similar to those recorded during the in vitro cavitation experiments were detected. We conclude that the electromagnetic probe is capable of detecting and quantifying aspects of cavitational activity in vitro, and potentially also in vivo. The observation that the probe responds directly to the motion of cavitation bubbles, coupled with its ability to detect cavitation at a distance, give it the potential for use in a range of medical and industrial applications. PMID- 10374989 TI - Effect of macroscopic air bubbles on cell lysis by shock wave lithotripsy in vitro. AB - In studies of cells or stones in vitro, the material to be exposed to shock waves (SWs) is commonly contained in plastic vials. It is difficult to remove all air bubbles from such vials. Because SWs reflect at an air-fluid interface, and because existing gas bubbles can serve as nuclei for cavitation events, we sought to determine in our system whether the inclusion of small, visible bubbles in the specimen vial has an effect on SW-induced cell lysis. We found that even small bubbles led to increased lysis of red blood cells (1- to 3-mm diameter bubbles, 9.8+/-0.5% lysis, n = 7; no bubbles, 4.4+/-0.8%, n = 4), and that the degree of lysis increased with bubble size. Damage could not be reduced by centrifuging the cells to the opposite end of the vial, away from the bubble. B-scan ultrasound imaging of blood in polypropylene pipette bulbs showed that, with each SW, bubbles were recruited from the air interface, mixing throughout the fluid volume, and these appeared to serve as nuclei for increased echogenicity during impact by subsequent SWs; thus, bubble effects in vials could involve the proliferation of cavitation nuclei from existing bubbles. Whereas injury to red blood cells was greatly increased by the presence of bubbles in vials, lytic injury to cultured epithelial cells (LLC-PK1, which have a more complex cytoarchitecture than red blood cells) was not increased by the presence of small air bubbles. This suggests different susceptibility to SW damage for different types of cells. Thus, the presence of even a small air bubble can increase SW induced cell damage, perhaps by increasing the number of cavitation nuclei throughout the vial, but this effect is variable with cell type. PMID- 10374990 TI - Ultrasound of radial, ulnar, median, and sciatic nerves in healthy subjects and patients with hereditary motor and sensory neuropathies. AB - This study was conducted to evaluate the capability of ultrasonography to visualize extremity nerves. Fifty healthy women and men and 10 patients suffering with hereditary motor and sensory neuropathy (HMSN) were examined. The radial nerve lateral to the humerus, ulnar nerve distal to the cubital tunnel, median nerve in the middle of the forearm and proximal to the palmar crease, sciatic nerve in the middle of the thigh and tibial and common peroneal nerves just distal to their bifurcation, were investigated, employing a 7.5-MHz electronic linear-array transducer. In healthy subjects, the median, ulnar and radial nerves could be identified in all cases, the sciatic nerve in 37 cases, the tibial and peroneal nerves in 10 cases. The median values of thicknesses were about 3 mm for arm nerves and 6 to 7 mm for the sciatic nerve. Nerve sizes did not correlate with subjects' height, weight or age. In the majority of the patients, arm and sciatic nerves were also visible. Thicknesses were normal in 34, increased in 11 and decreased in six of 51 nerves. In conclusion, ultrasonography allows reliable imaging of the major arm nerves and, occasionally, facilitates visualization of the sciatic, tibial and peroneal nerves in healthy subjects. Nerve size and structure did not differ significantly between patients with HMSN and healthy subjects. PMID- 10374991 TI - On clonal expansion and its effects on mutant frequencies, mutation spectra and statistics for somatic mutations in vivo. PMID- 10374992 TI - Failure to adequately use positive control data leads to poor quality mouse lymphoma data assessments. PMID- 10374993 TI - The mouse lymphoma assay in the wake of ICH4--where are we now? PMID- 10374994 TI - Issues for conducting the microtiter version of the mouse lymphoma thymidine kinase (tk) assay and a critical review of data generated in a collaborative trial using the microtiter method. PMID- 10374995 TI - Comparison of AluI-induced frequencies of dicentrics and translocations in human lymphocytes by chromosome painting. AB - It has been shown repeatedly that following irradiation of human lymphocytes in the G0 stage, more translocations are induced than dicentrics. To check the role of DNA double-strand breaks (DSB) alone for the induction of symmetrical and asymmetrical chromosome aberrations, the frequencies of induced exchange aberrations by the restriction enzyme AluI were analyzed. The enzyme was introduced into cells using the pellet pipetting technique. Frequencies of induced translocations and dicentrics were determined using a chromosome painting assay with chromosome-specific DNA libraries for chromosomes 1, 4 and X (representing 16.8% of the human genome). The number of translocations detected was approximately 3-fold higher than the number of dicentrics, indicating that the increased frequency of translocations compared with dicentrics found in irradiated human lymphocytes does not result from DNA lesions other than DSB but from differential processing of DSB. PMID- 10374997 TI - The isfA mutation specifically inhibits the SOS-dependent mutagenic pathway and does not selectively affect any particular base substitution. AB - We have previously described a new mutation in Escherichia coli, isfA, which causes inhibition of SOS mutagenesis (UV-induced in rec+ and spontaneous in recA730 strains) and several SOS-dependent phenomena. Antimutagenic activity of the isfA mutation in the recA730 strain was shown to be related to inhibition of processing of UmuD to UmuD' by RecA* coprotease. In the present study we have analysed the specificity of the antimutagenic activity of the isfA mutation by employing F' plasmids carrying a set of mutant lacZ genes that can individually detect two types of transitions and four types of transversions. Analysis revealed that isfA inhibits UV-induced G:C-->A:T and A:T-->G:C transitions, but does not affect the same G:C-->A:T transitions induced by EMS, an SOS-independent mutagen. Analysis of the antimutagenic activity of the isfA mutation in two mutator strains, recA730 and mutL, showed that isfA inhibits SOS-dependent transversions in recA730, but not transitions generated as replication errors in the mutL strain. In the double mutant recA730 mutL, both transitions and transversions were enhanced and isfA inhibits most transversions and only those transitions generated by the recA730 mutation. The results indicate that the antimutagenic activity of the isfA mutation is specific for the SOS, UmuD'C dependent mutagenic pathway but does not selectively affect any particular base substitution. Moreover, studies on the effect of the isfA mutation on transitions and transversions in different genetic backgrounds enable us to recognize different mutagenic pathways active in recA730 cells. PMID- 10374996 TI - Characterization of color mutants in lacZ plasmid-based transgenic mice, as detected by positive selection. AB - The plasmid-based transgenic mouse model, which uses the lacZ gene as the target for mutation, is sensitive to a wide range of in vivo mutations, ranging from point mutations to insertions and deletions extending far into the mouse genome. In this study, the nature of subtle lacZ mutations, which do not completely abolish beta-galactosidase activity, as detected by positive selection, was investigated. These subtle mutants are called 'color mutants' due to their light blue staining on X-gal medium. Replating of color mutants and retransformation of plasmid DNA, purified from individual color mutants, resulted in the same phenotype as the original color mutant. The p-gal positive selection system tolerates approximately 10% of wild-type activity as indicated by spectrophotometric determination of beta-galactosidase activity of individual color mutants. Restriction digestion and size separation of plasmid DNA revealed no visible change in the size of the plasmid in color mutants. Sequence analysis confirmed the presence of a point mutation in each lacZ gene of nine different color mutants. The results indicate that color mutants are caused neither by the presence of a mixture of wild-type and mutated lacZ plasmids within the same host cell nor by a mixture of cells within the original mutant colony which carry either wild-type or mutated lacZ plasmids. In addition, it was discovered that the mouse line studied harbors four polymorphic base changes among the integrated plasmid copies. PMID- 10374998 TI - Standardization and validation of DNA adduct postlabelling methods: report of interlaboratory trials and production of recommended protocols. AB - The aim of this project was to devise and test improved protocols of the 32P postlabelling assay for the detection of carcinogen-DNA adducts. The intention was to reverse the drift of different investigators using increasingly divergent experimental conditions. This would lead to a more standardized assay that can be used in future applications by different investigators for the monitoring of human exposure to genotoxic agents, permitting more meaningful comparisons between different studies or between different participants in the same study. As part of this process, there was perceived to be a need for carcinogen-modified DNA standards of known levels of adducts for use as positive controls, as standards for normalization of results with unknown samples and to assist interlaboratory comparisons. The preparation of characterized DNA standards modified by benzo[a]pyrene (BaP), a polycyclic aromatic hydrocarbon (PAH), 4 aminobiphenyl (ABP), an aromatic amine, 2-amino-1-methyl-6-phenylimidazo[4,5 b]pyridine (PhIP), a heterocyclic amine, and N-methyl-N-nitrosourea (MNU), a methylating agent yielding DNA containing O6-methylguanine, was carried out. A critical appraisal of all aspects of the 32P-postlabelling procedure and investigations to examine the influence of a number of key variations on the assay were conducted. There followed testing of a consensus protocol in a first interlaboratory trial involving 25 participants in Europe and the USA, conducted on the prepared synthetic DNA standards, the assessment of interlaboratory variability and the reasons for it. Revision of the protocols was followed by further testing in a second interlaboratory trial in which liver DNA from mice treated with BaP or ABP were assayed together with the synthetic DNA standards. Adduct levels were found to be significantly lower by 32P-postlabelling than by 3H incorporation. A recommended set of procedures has been developed for the detection and quantitation of DNA adducts formed by PAHs, aromatic amines and methylating agents. These trials have led to a much clearer idea as to what are the critical features and procedures of the 32P-postlabelling assay and there is a set of standard DNA samples for use in quality control and against which biological samples can be normalized. Use of these standards and procedures has reduced interlaboratory variability in quantitation of DNA adducts. PMID- 10374999 TI - Nucleotide excision repair modulates the cytotoxic and mutagenic effects of N-n butyl-N-nitrosourea in cultured mammalian cells as well as in mouse splenocytes in vivo. AB - The butylating agent N-n-butyl-N-nitrosourea (BNU) was employed to study the role of nucleotide excision repair (NER) in protecting mammalian cells against the genotoxic effects of monofunctional alkylating agents. The direct acting agent BNU was found to be mutagenic in normal and XPA mouse splenocytes after a single i.p. treatment in vivo. After 25 and 35 mg/kg BNU, but not after 75 mg/ kg, 2- to 3-fold more hprt mutants were detected in splenocytes from XPA mice than from normal mice. Using O6-alkylguanine-DNA alkyltransferase (AGT)-deficient hamster cells, it was found that NER-deficient CHO UV5 cells carrying a mutation in the ERCC-2 gene were 40% more mutable towards lesions induced by BNU when compared with parental NER-proficient CHO AA8 cells. UV5 cells were 1.4-fold more sensitive to the cytotoxic effects of BNU compared with AA8 cells. To investigate whether this increased sensitivity of NER-deficient cells is modulated by AGT activity, cell survival studies were performed in human and mouse primary fibroblasts as well. BNU was 2.7-fold more toxic for mouse XPA fibroblasts compared with normal mouse fibroblasts. Comparable results were found for human fibroblasts. Taken together these data indicate that the role of NER in protecting rodent cells against the mutagenic and cytotoxic effects of the alkylating agent BNU depends on AGT. PMID- 10375000 TI - Alternatives in the induction and preparation of phenobarbital/naphthoflavone induced S9 and their activation profiles. AB - With the aim of optimizing the efficiency of S9 fractions used in in vitro mutagenicity assays, different schemes for the induction of liver enzymes in rats were tried and the amount of S9 fraction required was assessed. The activity of 2 anthramine (2AA), 2-acetylaminofluorene (2AAF), 3-methylcholanthrene (3MTCL) and benzo[a]pyrene in bacterial mutagenicity tests was compared with the enzymatic activity in S9 fractions obtained from rats treated with either phenobarbital (NaPB), beta-naphthoflavone (betaNF) or combinations of both. Three pool systems prepared with different amounts of NaPB-induced S9 and betaNF-induced S9 were also analyzed for their activation capacities. Profiles of standard plate incorporation assays with Salmonella typhimurium TA98 increased with the amount of S9 fraction added for all drugs tested, except for 2AA, which showed a maximun of activity at low protein concentrations. According to these profiles, an optimal S9 protein content of 700-1000 microg/plate was estimated. For 2AAF and 3MTCL an S9 fraction obtained following a simultaneous treatment with NaPB (i.p.) and betaNF (oral gavage) (NaPB + betaNF) yielded the greatest response. This preparation was the only one which produced positive activation with 3MTCL as test drug. With the other test drugs all the S9 fractions were very active, including the NAPB + betaNF-induced S9. Both Phase I and Phase II cytochrome P450 enzymatic activities were enhanced in this S9 fraction. These results suggest that the simultaneous treatment (NaPB + betaNF) would be an adequate inducer for in vitro activation when used at 700-1000 microg protein/plate. PMID- 10375001 TI - Mitotic aberrations induced by carbaryl reflect tyrosine kinase inhibition with coincident up-regulation of serine/threonine protein phosphatase activity: implications for coordination of karyokinesis and cytokinesis. AB - The insecticide carbaryl and its metabolite 1-naphthol cause partial uncoupling of karyokinesis and cytokinesis in V79 Chinese hamster fibroblasts; karyokinesis is blocked in metaphase, the microtubules of the spindle depolymerize and the chromosomes and spindle remnants become displaced to the periphery of the cell. A high frequency of these disturbed cells elongate and a smaller fraction initiate a cleavage furrow. Here, we attempt to determine the potential targets for carbaryl and 1-naphthol in cytokinesis-specific signalling, led by the fact that the potential protein phosphatase inhibitor 1-naphthyl phosphate was previously identified in treated cells. We found that the typical cytological pattern induced by carbaryl and 1-naphthol could be obtained with tyrphostins, specific tyrosine kinase inhibitors, indicating that the carbaryl-induced effects could be due to tyrosine kinase inhibition. This was confirmed by tyrosine kinase assays showing that carbaryl, 1-naphthol and 2-naphthol were equally efficient at inhibiting tyrosine kinase activity as tyrphostin B44(-). As tyrosine kinases can act as regulatory factors in determining dephosphorylation rates, the activities of type-1 (PP1) and type-2A (PP2A) serine/threonine protein phosphatases were also determined. There was a clear up-regulation of the overall PP1/PP2A activities in cells treated with carbaryl, 1-naphthol or tyrphostin B44(-). This stimulation was shown to be indirect because these compounds had no effect on the activity of purified human PP1 in the test tube. 2-Naphthol, which has been found to be less efficient with regard to displacement of chromatin, did not cause up regulation, but a significant decrease in PP1/PP2A activity. We suggest that a net decrease in tyrosine kinase activity in combination with a net increase in PP1/PP2A activity is a precondition for cell elongation and cytokinesis in mammalian cells and that the corresponding enzymes are targets in the network of activities serving to coordinate karyokinesis and cytokinesis. PMID- 10375002 TI - Effects of oleic acid, docosahexaenoic acid and eicosapentaenoic acid on background and genotoxin-induced frequencies of SCEs in Indian muntjac fibroblasts. AB - Muntjac cells were cultured at 5 X 10(5) cells/10 cm Petri dish for 24 h prior to addition of fatty acids (50 microM) which were delivered to the cells complexed with 2% bovine serum albumin (fatty acid-free) and incubated for a further 24 h. Parallel dishes were processed for lipid extraction and GC analysis. This analysis showed highly significant (P < 0.01) uptake by the cells of each fatty acid. Genotoxins (75 microM hydrogen peroxide, 20 microM t-butylhydroperoxide and 2.4 microM mitomycin C) were added to the cells for 1 h prior to the end of the 24 h fatty acid incubation period. Control (no genotoxin or fatty acid) treatments were included. No difference was observed in background frequencies of SCEs between controls and fatty acid treatments, thus indicating that these fatty acids per se do not cause DNA damage. The cells incubated with the genotoxins showed increased (P < 0.05) frequencies of SCEs when compared with control frequencies. Cells incubated with genotoxins in the presence of fatty acids also showed significantly higher (P < 0.05) levels of SCEs when compared with control frequencies. When cells supplemented with genotoxins in the presence of fatty acids were compared with cells treated with genotoxins alone, higher levels of SCEs were observed in the former, suggesting that the fatty acids exacerbate DNA damage caused by these genotoxins. PMID- 10375004 TI - Stereoselective formation of glutathione S-conjugates from halovinylmercapturate sulphoxides. AB - 1. The glutathione S-transferase catalysed formation of glutathione S-conjugates from halovinylmercapturate sulphoxides was investigated in rat liver and kidney cytosol, with purified glutathione S-transferases and in rat in vivo. 2. The two diastereomers of the sulphoxides of N-acetyl-S-(1,2-dichlorovinyl)-L-cysteine, N acetyl-S-(2,2-dichlorovinyl)-L-cysteine and N-acetyl-S-(1,2,2-trichlorovinyl)-L cysteine show different reactivities with glutathione and glutathione S transferases. Rat liver and kidney cytosol catalyses the formation of a 1:1 mixture of two diastereomers of (E)-N-acetyl-S-(2-glutathione-S-yl-2-chlorovinyl) L-cysteine sulphoxide from N-acetyl-S-(2,2-dichlorovinyl)-L-cysteine sulphoxide and of (E)-N-acetyl-S-(2-glutathione-S-yl-1,2-dichlorovinyl)-L-cysteine sulphoxide from N-acetyl-S-(1,2,2-trichlorovinyl)-L-cysteine sulphoxide. In contrast, only one diastereomer of the Z-isomers was formed. 3. N-acetyl-S-(1,2 dichlorovinyl)-L-cysteine sulphoxide reacted spontaneously with glutathione at high rates, a 1:1 mixture of both diastereomers of N-acetyl-S-(2-glutathione-S-yl 1-chlorovinyl)-L-cysteine sulphoxide was formed. 4. Metabolism of N-acetyl-S-(2,2 dichlorovinyl)-L-cysteine sulfoxide and N-acetyl-S-(1,2,2-trichlorovinyl)-L cysteine sulfoxide under by alpha-class glutathione S-transferases yielded identical products as observed with the cytosolic enzymes. No reaction was observed in the presence of rat liver mu class glutathione S-transferases or human glutathione S-transferase M1. 5. Formation of these glutathione conjugates was also observed in the bile of rat after i.p. administration of the mercapturic acid sulphoxides. The results obtained show that stereochemical aspects may govern the regioselectivity and substrate specificity in glutathione S transferase-catalysed reactions. PMID- 10375003 TI - DNA repair methyltransferase (Mgmt) knockout mice are sensitive to the lethal effects of chemotherapeutic alkylating agents. AB - We have generated mice deficient in O6-methylguanine DNA methyltransferase activity encoded by the murine Mgmt gene using homologous recombination to delete the region encoding the Mgmt active site cysteine. Tissues from Mgmt null mice displayed very low O6-methylguanine DNA methyltransferase activity, suggesting that Mgmt constitutes the major, if not the only, O6-methylguanine DNA methyltransferase. Primary mouse embryo fibroblasts and bone marrow cells from Mgmt -/- mice were significantly more sensitive to the toxic effects of the chemotherapeutic alkylating agents 1,3-bis(2-chloroethyl)-1-nitrosourea, streptozotocin and temozolomide than those from Mgmt wild-type mice. As expected, Mgmt-deficient fibroblasts and bone marrow cells were not sensitive to UV light or to the crosslinking agent mitomycin C. In addition, the 50% lethal doses for Mgmt -/- mice were 2- to 10-fold lower than those for Mgmt +/+ mice for 1,3 bis(2chloroethyl)-1-nitrosourea, N-methyl-N-nitrosourea and streptozotocin; similar 50% lethal doses were observed for mitomycin C. Necropsies of both wild type and Mgmt -/mice following drug treatment revealed histological evidence of significant ablation of hematopoietic tissues, but such ablation occurred at much lower doses for the Mgmt -/- mice. These results demonstrate the critical importance of O6-methylguanine DNA methyltransferase in protecting cells and animals against the toxic effects of alkylating agents used for cancer chemotherapy. PMID- 10375005 TI - N-sulphoconjugation of amines by human cytosolic hydroxysteroid sulphotransferase. AB - 1. N- and O-sulphoconjugation of various substrates were studied with human liver cytosol and purified cytosolic sulphotransferase in the presence of 3 phosphoadenosine 5-phosphosulphate. 2. Human liver cytosol catalysed N sulphoconjugation of alicyclic and aryl-amines, and O-sulphoconjugation of hydroxysteroid and phenol. Activities of amine sulphoconjugation in the cytosol correlated well with those of hydroxysteroid but not with phenol. 3. Alicyclic amine sulphotransferase in human liver cytosol was purified to homogeneity by anion exchange, affinity and hydroxyapatite chromatography. Sulphoconjugating activities of alicyclic amine co-purified with those for hydroxysteroid conjugation. Subunit molecular weight of the purified sulphotransferase was 34 kDa. Contents of the purified enzyme correlated with the sulphoconjugating activities of hydroxysteroid and alicyclic amine. From these results, we concluded that the alicyclic amine sulphotransferase purified in this study was identical to hydroxysteroid sulphotransferse in human liver cytosol. 4. The results of this study indicate that hydroxysteroid sulphotransferase in human liver cytosol catalyses N-sulphoconjugation of alicyclic and aryl-amines. Hydroxysteroid sulphotransferase in the cytosol is reported to catalyse O sulphoconjugations of various compounds including hydroxysteroids, bile acids, cholesterol, and aliphatic and benzylic alcohols. The present and previously reported results indicate that hydroxysteroid sulphotransferase in the cytosol catalyses both N- and O-sulphoconjugations of several substrates. PMID- 10375006 TI - Intracellular localization and metabolism of the phenanthridinium trypanocide, ethidium bromide, by isolated rat hepatocytes. AB - 1. Confocal laser scanning microscopy (CLSM) has shown that ethidium (3 ,8 diamino-5-ethyl-6-phenylphenanthridinium) bromide, an aromatic phenanthridinium trypanocide, is taken up rapidly into the nucleoli and nuclear membranes of isolated rat hepatocytes. 2. It is biotransformed by the hepatocytes and at least five metabolites have been detected by high-performance liquid chromatography (HPLC). 3. Two new metabolites, 3- and 8-N-glucuronosylethidium, have been identified by HPLC-electrospray mass spectrometry and they represent the major pathway of metabolism, accounting for 6.4 +/- 0.7 and 19.5 +/- 1.2% respectively of total recovered drug after incubation. A third metabolite, 3,8 diacetylethidium, is formed in trace quantities. 4. The other two metabolites, 3 acetylethidium and 8-acetylethidium, have been reported previously. PMID- 10375007 TI - Molecular modelling of CYP2B6, the human CYP2B isoform, by homology with the substrate-bound CYP102 crystal structure: evaluation of CYP2B6 substrate characteristics, the cytochrome b5 binding site and comparisons with CYP2B1 and CYP2B4. AB - 1. Molecular modelling studies of CYP2B isoforms from rat (CYP2B1), rabbit (CYP2B4) and man (CYP2B6) are reported, with particular emphasis on substrate interactions with the human CYP2B isoform, CYP2B6. 2. The findings represent an advance on our previous study that focused primarily on the rat CYP2B isoform, CYP2B1, and involved homology modelling with substrate-free CYP102. 3. The current work utilizes the recently published substrate-bound CYP102 crystal structure as a template for construction of the CYP2B subfamily isoforms and shows, in particular, that known CYP2B6 substrate specificity and regioselectivity can be rationalized by putative active site interactions. PMID- 10375008 TI - Preclinical and in vitro assessment of the potential of D0870, an antifungal agent, for producing clinical drug interactions. AB - 1. D0870, an azole antifungal agent, produced dose-related increases in total cytochrome P450 and aldrin epoxidase when administered as 14 daily oral doses (0, 0.5, 2.5 and 12.5 mg/kg/day) to the male rat. Administered as single doses, D0870 increased pentobarbitone-sleeping time in a dose-related manner. 2. In human hepatic microsomal incubations, D0870 produced pronounced inhibition of CYP2C9 (tolbutamide hydroxylase) and, to a lesser degree, CYP3A4 (testosterone 6beta hydroxylase), but had more limited effects on CYP1A2, 2C19 and 2D6 activity. In comparison with ketoconazole, itraconazole and fluconazole, D0870 was the most potent inhibitor of CYP2C9 activity. It is predicted that D0870 may inhibit the in vivo clearance of CYP2C9 substrates by approximately 58%, thereby increasing their steady-state concentrations by 2.4 times, which would be of clinical significance for some compounds. 3. During incubation of [14C]-D0870 with cultured human hepatocytes for up to 72 h, two discrete metabolites (A and B) were formed. Formation of metabolite A was abolished by both quinidine and ketoconazole and is probably CYP3A4-mediated, whereas generation of metabolite B did not appear to be dependent on cytochrome P450. 4. D0870 has potential to produce both induction and inhibition of cytochrome P450 enzymes in man. PMID- 10375009 TI - Microsomal metabolism of N,N-diethyl-m-toluamide (DEET, DET): the extended network of metabolites. AB - 1. The aim was to set out to establish the complete network of metabolites arising from the phenobarbital-treated rat liver microsomal oxidation of N,N diethyl-m-toluamide (DEET). The products formed from DEET and all its subsequent metabolites were identified by HPLC retention times, UV spectroscopy, mass spectrometry and by comparison with authentic standards. 2. DEET (1a) produces three major metabolites, N-ethyl-m-toluamide (1b), N,N-diethyl-m (hydroxymethyl)benzamide (2a) and N-ethyl-m-(hydroxymethyl)benzamide (2b), and, at low substrate concentrations or extended reaction times, two minor metabolites, toluamide (1c) and N,N-diethyl-m-formylbenzamide (3a). 1b and 2a are primary metabolites and their formation follows Michaelis-Menten-type kinetics. At low DEET concentrations, ring methyl group oxidation is favoured; at saturation concentrations, methyl group oxidation and N-deethylation proceed at similar rates. The rate of formation of 2b decreases with increasing DEET concentration; 2b is therefore a secondary metabolite of DEET and DEET acts as a competitive inhibitor of the metabolism of 1b and 2a. 3. Except for the primary amides, where N-dealkylation is impossible, metabolism of all subsequent compounds, 1b,c, 2a-c, 3a-c and 4a,b, involves an N-deethylation (NEt2 --> NHEt or NHEt --> NH2) competitive with a ring substituent oxidation (CH3 --> CH2OH, CH2OH --> CHO or CHO --> CO2H). Surprisingly, the aldehydes 3a-c are also reduced to the corresponding alcohols 2a-c (CHO --> CH2OH); CO inhibits the oxidative metabolism of 3a-c, but reduction to 2a-c continues uninhibited. 4. The outcomes of this work are that (1) previously unreported aldehydes 3b and 3c form part of the DEET network of metabolites, (2) the reduction of the aldehydes 3a-c has the potential to inhibit the formation of the more highly oxidized DEET metabolites, (3) amide hydrolysis was not observed for any substrate and (4) no evidence was obtained for N-(1-hydroxyethyl)amide intermediates. PMID- 10375010 TI - Elimination of methoxyacetic acid and ethoxyacetic acid in rat. AB - 1. The pharmacokinetics of methoxyacetic acid (MAA) and ethoxyacetic acid (EAA) have been determined in the male and female rat following bolus intravenous administration at 100 mg/kg. The plasma-concentration data of MAA fitted well to a one-compartment model, and the plasma-concentration data of EAA to a two compartment model. 2. The elimination half-life of MAA estimated from plasma data was higher in females (18.6+/-2.0 h) than in males (13.2+/-0.4 h). There was no difference in the elimination half-lives estimated from urine data. The apparent volume of distribution was lower in the male than in the female rat estimated from plasma data only, while AUC, total and non-renal clearances and the relative amount MAA excreted unchanged in urine was similar in the male and female rat. 3. Clearance of EAA is higher than of MAA, and this appears as a result of metabolic capacity. The elimination half-lives of EAA were similar in the male and female rat, 9.4+/-3.7 and 10.5+/-2.6 h respectively. AUC was higher in the female compared with the male rat. The fraction of EAA eliminated during the distribution phase was 44.0+/-15.4 and 41.0+/-17.4% in the male and female rat respectively. The initial volume of distribution, the apparent volume of distribution, total and non-renal clearance are higher in the male compared with the female rat. PMID- 10375011 TI - Pharmacokinetics of a new histamine H2-receptor antagonist, Z-300, in rat and dog. AB - 1. The plasma level of Z-300 reached a maximum (Cmax) at 30 min after the oral administration of Z-300 to dog, and disappeared from the systemic circulation with a half-life of 8-9 h. The bioavailability of Z-300 was 52% after the oral administration of Z-300, 3 mg/kg. At doses ranging from 3 to 30 mg/kg, Cmax and AUC (area under the plasma concentration-time curve) were proportional to the dose. 2. The plasma level of Z-300 reached Cmax at 10 min after the oral administration of Z-300 to rat, and disappeared from the systemic circulation with a half-life of 0.8-1.6 h. The bioavailability of Z-300 was 39% after the oral administration of Z-300, 10 mg/kg, and there was a non-linear relationship between the plasma level-time profile of Z-300 and the administered dose (3-50 mg/kg). 3. The binding of Z-300 to plasma protein was 92% in man, 65% in dog and 25% in rat. It is suggested that these species differences were due to the content of alpha1-acid glycoprotein (alpha1-AG), because Z-300 bound more strongly to alpha1-AG than to albumin. PMID- 10375012 TI - Inhibition of protein phosphatase 2A (PP2A) mimics suckling-induced sensitization of mammotropes: involvement of a pertussis toxin (PTX) sensitive G-protein and the adenylate cyclase (AC). AB - In lactating rats, suckling renders mammotropes more responsive to prolactin (PRL)-releasing stimuli and less responsive to PRL-inhibiting secretagogues. We have previously shown that a decrease in the activity of protein phosphatase 2A (PP2A) may be responsible for the decrease in responsiveness to the inhibitory secretagogue dopamine (DA). In our present experiments, we have studied the involvement of the adenylate cyclase (AC), stimulatory and inhibitory GTP-binding proteins and also the role of PP2A in the sensitization phenomenon. Pituitary cells obtained from mother rats separated from their pups for 4 h prior to dispersion (non-suckled), suckled for 10 or 30 min after the separation period (suckled) and without separation (continual suckling) were incubated in the presence of different doses of forskolin to activate AC and DA. In a further study, pituitary cells of non-suckled rats were pretreated with cholera toxin (CTX) or pertussis toxin (PTX) and tested for the stimulatory action of forskolin or TRH on PRL release. Ocadaic acid (OA) pretreatment has been used to investigate the involvement of PP2A. Hormone secretion was measured by the reverse hemolytic plaque assay (RHPA). Our results have shown that cells from non suckled rats were unresponsive to forskolin. A 10-min suckling stimulus sensitizes pituitary mammotropes to respond with a PRL release to a dose dependent activation of AC by forskolin. This sensitization of AC becomes a permanent feature of the cells when suckling continues for an additional 20 min. We have also found that pituitary mammotropes from non-suckled dams respond to forskolin or TRH with PRL release when they were preincubated with either PTX or the PP2A inhibitor OA. It clearly indicates that the non-responsive pituitary can be shifted to the responsive stage by uncoupling of inhibitory G-protein from its receptor as well as by inhibition of PP2A. This latter finding, consonant with our previous results, suggests that suckling may cause selective changes in the function of G(i) of mammotropes due to a rapid phosphorylation which can remove tonic, GTP-dependent inhibitory function. PMID- 10375013 TI - Hormonal regulation of proliferation of granulosa and Leydig cell lines derived from gonadal tumors of transgenic mice expressing the inhibin-alpha subunit promoter/simian virus 40 T-antigen fusion gene. AB - We have produced a transgenic (TG) mouse model expressing the Simian Virus 40 T antigen (Tag) gene, driven by a 6-kb fragment of the mouse inhibin-alpha subunit promoter (inh-alpha). The mice develop gonadal tumors with 100% penetrance by the age of 5-8 months, of granulosa cell origin in the ovary, and of Leydig cell origin in the testis. In the present study, we characterized the hormonal regulation of proliferation of two immortalized cell lines, BLT-1, originating from a Leydig cell tumor, and NT-1, originating from a granulosa cell tumor. [3H] thymidine incorporation in both types of cells was stimulated by activin (> or = 10-30 microg/l), while inhibin had no effect. Transforming growth factor (TGF) beta, at > or = 0.01 microg/l, stimulated proliferation of the granulosa tumor cells, but no effect was found on the Leydig tumor cells. Progesterone inhibited the proliferation of both cell lines, although the granulosa tumor cells were clearly less sensitive than the Leydig cells to this effect ( > or = 3 micromol/l vs. > 10 nmol/l, respectively). hCG had no effect on the Leydig tumor cell DNA synthesis whereas at high concentration (100 microg/l) it stimulated that of the granulosa cells. We also investigated in BLT-1 and NT-1 cells whether the proliferative changes were related to concomitant changes in Tag expression. In BLT-1 cells, this was stimulated by activin, progesterone and hCG, even though the latter substance did not affect cell proliferation. In contrast, TGF-beta inhibited Tag expression. In NT-1 cells, the expression of Tag was stimulated by activin, while hCG had no effect. In contrast, it was reduced by progesterone, inhibin and TGF-beta. In conclusion, our results indicate that the granulosa and Leydig tumor cells, despite similar mechanism of immortalization, respond differently to several mitotic stimuli. The responses in the level of Tag expression in these cells did not always correlate with the changes observed in cell proliferation, indicating the independence of these two phenomena. PMID- 10375014 TI - Growth hormone suppression of insulin-like growth factor binding protein-1 promoter activity. AB - Hepatic transcription of insulin-like growth factor binding protein-1 (IGFBP-1) is rapidly downregulated by growth hormone (GH) which is also known to induce expression of c-fos and c-jun. Co-expression of c-fos or c-jun in rat hepatocytes, individually or together, suppresses IGFBP-1 promoter activity by approximately 60%. When hepatic nuclear extracts from sham-operated, hypophysectomized (hypox) and GH-treated hypox rats were analyzed by DNase-1 footprinting, differences in the protection pattern were identified in three regions of the IGFBP-1 promoter. F1 corresponding to - 660 to - 640 bp showed acute changes in response to GH administration. In additional regions, F2 and F3, representing - 758 to - 748 bp and - 477 to - 447 bp, respectively, differences were apparent between nuclear extracts from the hypox and sham-operated rats. When F1 and F2 were removed by deletion of the region from - 824 to - 557 bp, the GH response was lost but suppression by co-expression of c-fos and c-jun was preserved. A putative AP-1 binding site was present in the F3 footprint region, however removal of F3 did not affect the GH responsiveness. These data indicate that several distinct sequences, other than the putative AP-1 site are involved in mediating the GH effects on IGFBP-1 transcription. PMID- 10375015 TI - Vascular endothelial growth factors are differentially regulated by steroid hormones and antiestrogens in breast cancer cells. AB - Vascular endothelial growth factor (VEGF) is a major inducer of tumor angiogenesis and an important prognostic factor in breast cancer. Hypoxia is an important inducer of VEGF expression but less is known of the role of hormones in VEGF regulation. We have studied the regulation of VEGF, VEGF-B, VEGF-C, and VEGF D mRNAs in human MCF-7 and mouse S115 breast carcinoma cells stimulated by estrogens and androgens, respectively. VEGF, VEGF-B, and VEGF-C were expressed in both cell lines, whereas VEGF-D was expressed only in S115 cells. Addition of estradiol (E2) caused a biphasic increase of VEGF mRNA in MCF-7 cells and led to accumulation of the VEGF protein in the culture medium. The VEGF-B mRNA was not affected, while a decrease occurred in VEGF-C mRNA. Similarly, testosterone upregulated the expression of VEGF mRNA in the S115 cells. Experiments with actinomycin D and cycloheximide suggested that estrogen induction of VEGF mRNA is dependent on the synthesis of new mRNA and increased mRNA half-life. The antiestrogen ICI 182.780 inhibited E2 stimulation of VEGF, suggesting that the effect was mediated by the estrogen receptor. In contrast, the antiestrogens tamoxifen and toremifene which inhibit MCF-7 cell growth in vivo and in vitro did not inhibit estrogen effect but induced VEGF mRNA expression when used alone. The antiandrogen cyprosterone acetate inhibited T induction of VEGF mRNA in S115 cells, thus suggesting that activation of androgen receptor must be involved in the increase of VEGF mRNA. Our results suggest that both estrogen and androgen stimulate the expression of VEGF by increasing gene transcription and mRNA stability. In addition, the antiestrogens tamoxifen and toremifene also increased VEGF expression. Estrogen and androgen induction of VEGF expression and promotion of new vessel formation may be an important paracrine mechanism by which these hormones contribute to the early phase of tumor growth of hormonal cancer. PMID- 10375016 TI - IGF-1 or insulin, and the TSH cyclic AMP cascade separately control dog and human thyroid cell growth and DNA synthesis, and complement each other in inducing mitogenesis. AB - The regular doubling of cell mass, and therefore of cell protein content, is required for repetitive cell divisions. Preliminary observations have shown that in dog thyrocytes insulin induces protein accumulation but not DNA synthesis, while TSH does not increase protein accumulation but triggers DNA synthesis in the presence of insulin. We show here that EGF and phorbol myristate ester complement insulin action in the same way. HGF is the only factor activating both protein accumulation and DNA synthesis. The effects of insulin on protein accumulation and in permitting the TSH effect are reproduced by IGF-1 and are mediated, at least in part by the IGF-1 receptor. The concentration effect curves are similar for both effects. Similar results are obtained in human thyrocytes. They reflect true cell growth, as shown by increases in RNA content and cell size. Carbachol and fetal calf serum also stimulate protein synthesis and accumulation without triggering DNA synthesis, but they are not permissive for the mitogenic effects of TSH or of the general adenylate cyclase activator, forskolin. Moreover the mitogenic effect of TSH greatly decreased in cells deprived of insulin for 2 days although these cells remain hypertrophic. Hypertrophy may therefore be necessary for cell division, but it is not sufficient to permit it. Three different mechanisms can therefore be distinguished in the mitogenic action of TSH: (1) the increase of cell mass (hypertrophy) induced by insulin or IGF-1; (2) the permissive effect of insulin or IGF-1 on the mitogenic effect of TSH which may involve both the increase of cell mass and the induction of specific proteins such as cyclin D3 and (3) the mitogenic effect of the TSH cyclic AMP cascade proper. PMID- 10375017 TI - Direct non-genomic effect of steroid hormones on superoxide anion generation in the bone resorbing osteoclasts. AB - We have investigated the possible acute effect of steroid hormones, including 1alpha,25 dihydroxycholecalciferol (1alpha,25(OH)2D3) and estradiol, on the generation of superoxide anion (O2*-) in bone resorbing osteoclasts. Evidence is presented demonstrating acute non-genomic stimulatory action of 1alpha,25(OH)2D3 on the production of free radicals by rat osteoclasts cultured on calcified matrix. The increase in O2*- production was observed in the range of 6-10 s (n = 5) following exposure of enriched osteoclasts to 1alpha,25(OH)2D3 and was found to be transient with the peak response being in the range of 5-45 s (n = 5). The decline in the transient was much slower than the elevation, time for the decay being in the range 1-5 min (n = 5) and remained above the levels present prior to the addition. The exposure of the osteoclast to dexamethasone was found to have no effect on O2*- generation, whilst estradiol was found to be inhibitory. The mode of stimulation and the kinetics of the transients of O2*- in the bone resorbing osteoclasts produced by 1alpha,25(OH)2D3 were similar to that of parathyroid hormone (PTH) and pertussis. The exposure of the bone resorbing osteoclasts to cholera toxin was found to have no effect, suggesting that the stimulatory action is unlikely to be mediated via cAMP elevation. The importance of these observations is discussed in the context of calcium homeostasis and bone physiology. PMID- 10375018 TI - Ligand regulation of juvenile hormone binding protein mRNA in mutant Manduca sexta. AB - Insect hemolymph juvenile hormone binding protein (hJHBP) regulates peripheral titers of its ligands, the juvenile hormones. In larvae of the black (bl) strain of the tobacco hornworm, Manduca sexta, treatment with small doses of juvenile hormone I (JH I) can also regulate titers of hJHBP. To further investigate this regulation, responsiveness of hJHBP mRNA expression to JH I was characterized in vivo. RNA analyzes revealed that transcript levels in fat body, the site of hJHBP synthesis, increased fivefold within several hours of treatment with physiological doses of hormone and remained elevated for approximately 16 h. Sensitivity to JH treatment was found to vary temporally. To ensure transcript identity, a wild-type cDNA clone and a bl RT-PCR fragment were sequenced and found to be 99% homologous. Together, these results suggest that JH participates in regulating expression of its transport protein in bl larvae by modifying the in vivo abundance of hJHBP's mRNA transcript. PMID- 10375020 TI - Two binding sites for thyroid transcription factor 1 (TTF-1) determine the activity of the bovine thyroglobulin gene upstream enhancer element. AB - A thyroid-specific enhancer element located upstream from the bovine thyroglobulin gene had been shown to contain three contiguous regions that are protected by thyroid transcription factor 1 (TTF-1) in footprinting experiments in vitro. The functional relevance of the individual TTF-1 binding sites was investigated in a transient assay in primary cultured thyrocytes. Using reporter constructs containing synthetic oligonucleotides overlapping the protected sequences we were able to show that only two out of the three TTF-1 binding sites exhibit transcription enhancing activity. Within the context of the complete enhancer sequence, the central 'inactive' TTF-1 site could be deleted whithout any consequence on the activity of the enhancer in the assay, whereas the presence of both terminal 'active' TTF-1 sites had previously been shown to be strictly required for enhancer function. Our results thus show that the activity of the bovine thyroglobulin upstream enhancer relies on the presence of a pair of TTF-1 binding sites separated by about 30 bp. These results also emphasize the need to assess experimentally the functional relevance of TTF-1 binding sites identified in footprinting experiments. PMID- 10375019 TI - Control of FSH receptor mRNA expression in rat granulosa cells by 3',5'-cyclic adenosine monophosphate, activin, and follistatin. AB - FSH is required to maintain FSH and LH/hCG receptors at elevated steady-state levels after receptor induction. Although this function of FSH is mediated by cAMP, how cAMP level is related to the maintenance of gonadotropin receptors is unknown. To investigate cAMP's effect on changes in the levels of FSH receptor mRNAs in rat granulosa cells, total RNA from cells was prepared and analyzed by Northern blots. Incubation with 8-Br-cAMP for 24 h produced a dose-related increase in FSH receptor mRNA in granulosa cells of DES-primed immature rats. On the other hand, 8-Br-cAMP, washed at 24 h, exerted inverse dose-related effects on FSH receptor mRNA levels at 96 h. The addition of 1 mM cAMP resulted in higher levels of FSH receptor mRNA than that induced by 0.2 mM cAMP at 24 h, while 0.2 mM cAMP is as effective as 1-2 mM cAMP for the induction of FSH receptor mRNA at 96 h. To further analyze cAMP's role in the production of activin in granulosa cells, we measured activin levels in the culture medium after the addition of 8 Br-cAMP. The levels of activin A were suppressed by the addition of 8-Br-cAMP in a dose-dependent manner. In addition, the procedure by which 8-Br-cAMP was removed after 24 h incubation showed that the level of activin in the medium increased after medium change. With regard to the actions of activin A on gonadotropin receptors, our laboratory has demonstrated that activin A increases the levels of FSH receptor mRNAs. Therefore, cAMP has a negative effect on FSH receptor expression by suppressing the activin level. Since follistatin production is up-regulated by cAMP in this system, we examined the effect of follistatin on FSH receptor mRNA level, which is induced by activin and FSH. Cotreatment with follistatin (0-100 ng/ml) and activin (50 ng/ml) in the presence of FSH (30 ng/ml) caused a significant reduction in FSH receptor mRNA levels induced by activin. Based on these observations, it is possible that cAMP has both stimulatory and inhibitory effects on the expression of gonadotropin receptors, and the overall influence of cAMP on their expression might be determined by the integration of such opposing effects. PMID- 10375021 TI - Regulation of prolactin receptor glycosylation and its role in receptor location. AB - In unstimulated mammary epithelial cells from virgin mice, the prolactin receptor exists as two isoforms: a 78 and a 70 kDa species. Both proteins are reduced to a single 61 kDa molecule after N-glycanase F treatment, indicating that their size difference is solely a result of carbohydrate content. Membrane fractionation experiments reveal that the smaller species is exclusively intracellular, while the larger one is located on the cell surface. Nitric oxide (NO) stimulates the migration of prolactin receptors from an internal pool to the plasmalemma in only 30 min and this redistribution is associated with an increase in molecular weight. Redistribution is blocked by swainsonine, but not by castanospermine or 1 deoxymannojirimycin, suggesting that the glycosylation step involved with translocation is either alpha-mannosidase II or N-acetylglucosamine (NAG) transferase I. The former is unaffected by NO but the activity of the latter is doubled 30 min after exposure to NO. These data suggest that prolactin receptors are retained intracellularly because of their incomplete N-glycosylation and that NO triggers their redistribution by stimulating the completion of this process, in part by increasing NAG transferase I activity. PMID- 10375022 TI - Expression of human estrogen receptor using an efficient adenoviral gene delivery system is able to restore hormone-dependent features to estrogen receptor negative breast carcinoma cells. AB - Estrogen receptor (ER)-negative breast carcinomas are often difficult to treat as they do not respond to hormone therapy. In an attempt to determine if expressing the human estrogen receptor in an ectopic manner could restore the hormone responsiveness of these cells, we have expressed the human ER in ER-negative MDA MB 231 breast cancer cells using a recombinant adenovirus gene delivery system that allows high level expression of ER in essentially all cells. In these cells, the ER was correctly translated, had a wild type hormone binding affinity (Kd = 0.6 nM), bound well to estrogen response element-containing DNA, and showed an activation pattern of estrogen response element-reporter gene activity by estrogen and antiestrogens very similar to that observed in MCF-7 breast cancer cells containing endogenous ER (stimulation by estrogen, no stimulation by the antiestrogens trans-hydroxytamoxifen or ICI 164384, and blockade of estradiol stimulation by trans-hydroxytamoxifen or ICI 164384). Intriguingly, estradiol stimulation of these cells was also able to induce expression of pS2, an estrogen regulated gene considered to be a favorable prognostic marker for endocrine therapy in ER-positive breast cancer cells. Expression of the ER had no effect by itself on the proliferation rate of MDA-MB 231 cells. However, treatment of the ER-containing cells with estradiol or with the pure antiestrogen ICI 164 384 suppressed proliferation of the cells while the antiestrogen trans hydroxytamoxifen had little effect on proliferation; and cotreatment with trans hydroxytamoxifen reversed the estradiol- or ICI 164 384-evoked suppression of proliferation. To understand the mechanism underlying the inhibition of proliferation by estradiol, we examined the expression of several growth related endogenous genes. c-Myc protooncogene expression was strongly inhibited by treatment with estradiol as was expression of BRCA1 and BRCA2 genes, which is in agreement with their mitogenic-dependent expression, while expression of beta actin, a housekeeping gene, was not affected by hormone treatment. Together, these data suggest that reexpressing the human ER in breast cancer cells that no longer express this protein renders them sensitive to hormone treatment. The ability of the antiestrogen ICI 164 384 to suppress the proliferation of ER negative breast cancer cells that reexpress ER might be useful ultimately as an endocrine gene therapy approach for controlling the growth of ER-negative breast cancer cells. The application of recombinant adenoviruses expressing the human ER presents interesting features which might be used as a basis for designing more powerful and effective treatments for ER-negative breast cancers. PMID- 10375023 TI - Dominant negative activity of thyroid hormone receptor variant alpha2 and interaction with nuclear corepressors. AB - The splicing variant of the thyroid hormone receptor alpha (TRalpha) gene, TR variant alpha2 (TRv alpha2), lacks the second half of the ninth heptad, a domain thought to be important for heterodimerization with retinoid X receptors (RXRs). In transient transfection studies, TRv alpha2 exhibits weak dominant negative inhibition of TRalpha1-mediated transcription. In contrast, a TRv alpha2 mutant in which the ninth heptad was restored (alpha2 + 9H), exhibits very strong dominant negative activity. We have examined the role of nuclear corepressors (CoRs) in the dominant negative activity of TRv alpha2 and alpha2 + 9H. Glutathione S-transferase pull down experiments revealed that TRv alpha2 barely interacts with CoRs, whereas alpha2 + 9H interaction with CoRs is as strong as that of TRalpha1. A P160R CoR box mutation was introduced in the context of TRv alpha2 and alpha2 + 9H, which nearly abolishes the ability of these receptors to interact with CoRs. In transient transfection the dominant negative activity of TRv alpha2 was only marginally impaired by the P160R mutation. In contrast, alpha2 + 9H-P160R had approximately 66% less dominant negative activity than alpha2 + 9H. These results suggest that the weak dominant negative activity of TRv alpha2 is due in part to its lack of interaction with CoRs, and that restoration of the ninth heptad restores CoR interaction and strong dominant negative activity. Further, the data reveal aspects of the dominant negative action that are dependent on the orientation of the TRE. PMID- 10375024 TI - The rat intraovarian interleukin (IL)-1 system: cellular localization, cyclic variation and hormonal regulation of IL-1beta and of the type I and type II IL-1 receptors. AB - An increasing body of evidence supports the possibility that intraovarian interleukin (IL)-1 plays an intermediary role in the periovulatory cascade. To gain further insight into the intraovarian IL-1 hypothesis, we studied the cellular localization cyclic variation and hormonal regulation of IL-1beta, as well as of the type I and type II IL-1 receptors (IL-1R) in immature rats. In situ hybridization localized IL-1beta and type I IL-1R transcripts to the granulosa cell compartment, the innermost layers of the theca interna and to the oocyte of the untreated immature ovary. Molecular probing of whole ovarian material in the course of a simulated estrous cycle revealed a progressive preovulatory increase in IL-1beta and type I IL-1R transcripts to an in vivo peak at the time of ovulation (3.0- and 2.5-fold increases over untreated controls; P < 0.05). Comparable efforts to localize and probe for type II IL-IR transcripts failed to elicit a detectable signal. The basal in vitro expression pattern of IL 1beta and type II IL-1R transcripts by whole ovarian dispersates revealed an early (4 h) spontaneous increase to a peak (2.1- and 5.8-fold increases over time 0: P < 0.05) followed by a gradual decline to a 48 h nadir. Treatment of whole ovarian dispersates with the IL-1 receptor antagonist (IL-1RA) or with IL-1beta failed to alter the initial (4 h) burst of IL-1beta or of type II IL-1R expression thereby suggesting IL-1-independence. Treatment with hCG proved equally ineffective. However, longer-term treatment of whole ovarian dispersates with IL-1beta produced a significant secondary increase (5.9-fold over time 0; P < 0.05) in IL-1beta (but not type II IL-1R) transcripts by 48 h. This IL-1 effect was completely blocked by co-treatment with IL-1RA thereby suggesting mediation via a specific IL-1 receptor. Qualitatively comparable but quantitatively reduced results obtained for isolated granulosa cells. The basal in vitro expression pattern of type I IL-1R transcripts by whole ovarian dispersates revealed a progressive spontaneous increase (3.1-fold increase overall) over the 48 h culture. Treatment with IL-1beta produced a significant (P < 0.05) increase (5 fold) in type I IL-1R transcripts by 48 h, an effect which was completely blocked by co-treatment with IL-1RA. Taken together, these observations: (1) localize IL 1beta and its type I receptor to granulosa cells, the innermost layers of the theca interna and to the oocyte; (2) confirm their periovulatory in vivo expression pattern; (3) document their expression by untreated cultured whole ovarian dispersates; and (4) demonstrate their in vitro responsiveness to receptor-mediated/IL-1-driven autocrine amplification. The type II IL-1R was undetectable in vivo, its in vitro expression pattern proving IL-1- and hCG independent. The periovulatory expression pattern of IL-1beta and its receptor (type I) is compatible with the notion that the intraovarian IL-1 system may play an intermediary role in the ovulatory process. PMID- 10375025 TI - Transgene and host growth hormone gene expression in pituitary and nonpituitary tissues of normal and growth hormone transgenic salmon. AB - Growth hormone (GH) gene expression has been examined in control and transgenic coho salmon containing a transgene comprised of the sockeye salmon GH1 gene under the control of the MT-B promoter from the same species. This transgene dramatically enhances the growth of salmonids, and raises serum GH levels some forty-fold. Transcript levels from this transgene were detected by RT-PCR using construct-specific GH primers in all tissues examined (liver, kidney, skin, intestine, stomach, muscle, spleen, pyloric caeca), and ranged from 0.1 - 9.4 pg/50 microg total RNA in different tissues as estimated by dot blot analysis. Interestingly, GH gene expression was also observed in intestine of control coho salmon by RT-PCR capable of detecting host and transgene transcripts using general primers. Sequence analysis of the intestinal GH mRNA from controls indicated it was derived from the coho GH2 gene. GH mRNA abundance analyzed by northern analysis indicates lower levels are found in large (400-500 g) than small transgenic salmon (20-21 g). No molecular evidence for transgene expression was obtained in tissues from transgenic fry, despite an obvious increase in size relative to control siblings, suggesting very low levels of transgene expression early in development. GH mRNA levels (per microg RNA) were also examined in the pituitary gland, and were found to be significantly lower (P < 0.01) in transgenic coho compared to nontransgenic animals of the same size. Pituitary glands of transgenic animals were also smaller than control animals of the same size, and pituitary size, expressed as a proportion of body weight, decreased with body size in transgenic but not control animals. These results imply that pituitary GH expression is regulated by negative feed-back controls as occurs in other vertebrate systems. GH mRNA was examined in pituitary glands by whole-mount in situ hybridization, and, whereas overall levels appeared reduced in transgenic animals, the site of hybridization did not differ between transgenic and control glands. PMID- 10375026 TI - Nicotinamide potentiates TSHR and MHC class II promoter activity in FRTL-5 cells. AB - Here we show that nicotinamide modulates the promoter activity of rat thyrotropin (TSHR) and major histocompatibility complex (MHC) class II genes in rat FRTL-5 thyroid cells, and have identified a novel mechanism for its action. TSHR and MHC class II, are potentiated through reduced expression of a common repressor of these two genes, TSEP-1 (TSHR suppressor element binding protein-1)/YB-1. Thus we show that TSHR mRNA is increased and TSHR promoter activity was concentration dependently activated from 0 to 40 mM nicotinamide. The promoter lengths of TSHR and MHC class II containing TSEP/YB-1 binding sites were enhanced by 40 mM nicotinamide, but not the ones deleted of these binding sites. TSEP-1/YB-1 binding to the recognition sites in both TSHR and MHC class II promoters was reduced in nicotinamide-treated FRTL-5 nuclear extracts. Nicotinamide reduced the expression of TSEP-1/YB-1 mRNA and TSEP-1/YB-1 protein in the nucleus. PMID- 10375027 TI - Ovarian estrogen receptor alpha and beta mRNA expression: impact of development and estrogen. AB - We tested the hypothesis that ERalpha and ERbeta mRNAs in the rat ovary are regulated during the post-natal period and in immature rats in response to estrogen treatment. Total ovarian ERbeta mRNA was more abundant than ERalpha mRNA and expression of ERbeta increased between post-natal days 4 and 12, coinciding with advancing folliculogenesis and an increase in granulosa cell numbers. In contrast, ERalpha mRNA levels remained relatively constant during this period. In situ hybridisation studies localised both ERalpha and ERbeta to granulosa cells of growing follicles, in 25 day old ovaries, although not all granulosa cells in a follicle or all follicles expressed the ERs. Diethylstilboestrol (DES) administered in vivo to 21 day old rats, for up to 4 days, did not significantly alter the expression of either ER as determined by RT-PCR, despite a 5.5-fold increase in granulosa cell number in these ovaries. In situ hybridisation studies established that DES-treatment down-regulated granulosa cell ER mRNAs. RT-PCR analyses on isolated granulosa cells confirmed that ERalpha was significantly down-regulated by DES. The predominance of ERbeta over ERalpha in the ovary and the regulation of ERbeta mRNA expression during ovarian development, is consistent with an important biological role for ERbeta in granulosa cell proliferation and differentiation. PMID- 10375028 TI - Prolactin receptors are expressed and hormonally regulated in rat Sertoli cells. AB - In this study, the protein and mRNA expression of the short and long forms of Prolactin (PRL) receptors (PRL-R) were examined by means of Northern and Western blotting analyses in rat testicular Sertoli cells. Transcripts for the short and long forms of PRL-R were detected with specific probes, five major mRNA species of about 1.9, 2.6, 3.0, 3.7 and 5 kb for the short form and two of about 10 and 1.3 kb for the long form. Under reducing conditions, the use of a specific antibody for the short form revealed a major molecular species of approximately 45 kDa. Two groups of molecular species were detected for the long form, several bands with high molecular masses (110-300 kDa) and others about 45-60 kDa. Finally, the expression of the long form of PRL-R was shown to be hormonally regulated as it was inhibited by follicle stimulating hormone (FSH) (ED50 = 5 ng/ml). Together, the localisation of PRL receptors to Sertoli cells as well as the regulatory action of FSH on these receptors suggest that PRL and or (a) PRL like activity(ies) might be considered as (a) potential regulator(s) of spermatogenesis. PMID- 10375029 TI - Metabolism of 1alpha,25(OH)2D3 and its 20-epi analog integrates clonal expansion, maturation and apoptosis during HL-60 cell differentiation. AB - Induction of growth arrest and monocyte differentiation of HL-60 leukemia cells by 1alpha,25 dihydroxyvitamin D3 (1alpha,25(OH)2D3) is well established. By contrast, we have observed, that 1alpha,25(OH)2D3 and its metabolites play separate roles in clonal expansion and survival of differentiating HL-60 cells. Cells that had differentiated by 48 h (CD14 positive) grew slower than control cells, whereas CD14 negative cells were growing faster at this time point. Inhibiting 1alpha,25(OH)2D3 or 1alpha,25(OH)2-20-epi-D3 metabolism, by the 25(OH)D3-24-hydroxylase inhibitor ketoconazole, abolished hyperproliferation of CD14 negative cells. Instead, both the onset of differentiation and subsequent apoptosis were enhanced. These events were associated with immediate up regulation of the cyclin-dependent kinase inhibitor p21(waf1) and a lack of sustained expression, respectively. Stimulation and inhibition of growth by vitamin D3-related compounds was observed to be concentration and metabolite specific. Low amounts of 1alpha,25(OH)2-20-epi-D3 and 1alpha,24,25(OH)3-20-epi-D3 stimulated HL-60 cell growth. At higher concentrations, 1alpha,25(OH)2-20-epi-D3 was a more potent inducer than 1alpha,24,25(OH)3-20-epi-D3 of HL-60 differentiation; 1alpha,25(OH)2-20-epi-24-oxo-D3 was exclusively pro differentiative at all concentrations. 1alpha,25(OH)2-20-epi-D3 and 1alpha,24,25(OH)3-20-epi-D3 stimulated proliferation of KG-1a leukemia cells, but neither of these compounds nor 1alpha,25(OH)3-20-epi-24-oxo-D3 exerted pro differentiative effects on these cells. These findings shed new light on the pro- and anti-proliferative effects of 1alpha,25(OH)2D3 and lead to the postulate that metabolism of 1alpha,25(OH)2D3 and its 20-epi analog regulates different subsets of genes so as to co-ordinate population expansion and the differentiation process. Furthermore, 1alpha,25(OH)2D3 metabolism and/or sensitivity to the effects of metabolites may be altered in transformed cells to derive a clonal advantage. PMID- 10375030 TI - Parathyroid receptor gene expression by epiphyseal growth plates in rickets and tibial dyschondroplasia. AB - PTH/PTHrP receptor gene expression was evaluated in situ in avian epiphyseal growth plates taken from normal, rachitic and tibial dyschondroplasia (TD) afflicted chicks induced by thiram or by genetic selection. In the normal growth plates, PTH/PTHrP receptor gene expression was localized to the maturation zone as demonstrated by the expression of collagen type II (col II), osteopontin (OPN) genes and alkaline phosphatase activity (AP). In TD, either induced by thiram or by genetic selection, normal levels of PTH/PTHrP receptor gene expression were observed up to 21 days post-hatch. In rickets, on the other hand, no PTH/PTHrP receptor gene expression was observed in the growth plate from day 8 of a vitamin D-deficient diet. In cultured chondrocytes, PTH caused time-dependent down regulation of its own receptor. These results suggest that alterations in the PTH/PTHrP receptor gene expression are associated with rickets but not with TD. The reduction in the PTH/PTHrP receptor gene expression in rickets may be due to the high plasma levels of PTH. PMID- 10375031 TI - Identification of an important thyrotrophin binding site on the human thyrotrophin receptor using monoclonal antibodies. AB - A thyrotrophin (TSH) binding site has been identified on the extracellular domain of the human thyrotrophin receptor (hTSHR) using monoclonal antibodies that recognise the native hTSHR. These antibodies were produced by immunising BALB/c mice with denatured recombinant material, selected by their reaction with recombinant hTSHR expressed on heterologous cell lines using flow cytofluorimetric analysis, and characterised by immunoblotting and immunoprecipitation. The epitopes the monoclonal antibodies recognise were determined using multiple overlapping synthetic peptides. All of the antibodies reacted with epitopes within the region 335-390; these epitopes must be accessible on the external surface of the native hTSHR. None of the antibodies stimulated cAMP production of recombinant hTSHR cell lines. The epitopes of two antibodies (residues 337-342 and 355-358) are in the small peptide thought to be removed by proteolytic processing of hTSHR. A further five different antibodies (determined from their variable region sequences) all reacted with residues 381 384 emphasising the immunogenicity of this region. The functional importance of residues 381-384 as a TSH binding site was shown by the fact that some of these monoclonal antibodies caused inhibition of radiolabelled TSH binding of 80-90% at 1 microg/ml and greater than 50% inhibition at 0.1 microg/ml (0.65 nM--i.e. comparable in effectiveness with TSH itself). Residues 381-384 may form part of the target regions recognised by inhibitory autoantibodies found in Graves' disease. PMID- 10375032 TI - The glucocorticoid properties of the synthetic steroid pregna-1,4-diene-11beta-ol 3,20-dione (deltaHOP) are not entirely correlated with the steroid binding to the glucocorticoid receptor. AB - The natural steroid 11beta-hydroxyprogesterone is not only a modulator of 11beta hydroxy-steroid dehydrogenase activity, but also an efficient inducer of tyrosine aminotransferase activity in hepatocytes. In contrast with the low affinity for the mineralocorticoid receptor. 11beta-hydroxyprogesterone binds well to both the glucocorticoid receptor and the carrier protein transcortin. It is accepted that the introduction of a 1:ene double bond into 3-keto 4:ene steroids increases the glucocorticoid potency, so that 3-keto-1,4:diene steroids show improved chemical stability and are more potent glucocorticoids than their respective 4:ene analogs. The steroid pregna-1,4-diene-11beta-ol-3,20-dione (deltaHOP) had previously been described as an anti-inflamatory compound and an inhibitor of macromolecular biosynthesis in thymocytes and lymphocytes. In such studies, deltaHOP also exhibited some particular glucocorticoid properties which made it attractive as a tool for the study of the mechanism of action of glucocorticoids. In the present paper we show that deltaHOP possesses some classical biological actions of glucocorticoids such as deposition of glycogen in rat liver, induction of TAT activity in hepatocytes, and inhibition of the uptake of leucine and thymidine by thymocytes. It also exhibits minimal sodium-retaining properties. Consistent with these biological effects, deltaHOP shows a 70 times lower relative binding affinity for the mineralocortioid receptor than aldosterone, but a reasonable affinity for the glucocorticoid receptor, and is as efficient as dexamethasone in dissociating the 90 kDa heat shock protein from the glucocorticoid receptor heterocomplex. However, the inhibition of the uptake of amino acids and nucleotides observed in the presence of deltaHOP is not efficiently blocked when thymocytes are coincubated in the presence of steroids with known antiglucocorticoid activity. deltaHOP is similarly inefficient in inducing chloramphenicol-acetyl transferase activity in cells transfected with a plasmid that possesses two canonical glucocorticoid-responsive elements. Unlike most glucocorticoids, deltaHOP does not induce the fragmentation of DNA in a regular pattern characteristic of apoptosis and it does not reduce thymus weight. This unusual dissociation of glucocorticoid parameters makes deltaHOP a useful tool to discriminate between mechanisms of action by which steroids can exert their biological effects. PMID- 10375033 TI - Medaka (Oryzias latipes) FTZ-F1 potentially regulates the transcription of P-450 aromatase in ovarian follicles: cDNA cloning and functional characterization. AB - Our previous findings suggest the activity of cytochrome P-450 aromatase (P 450arom), the enzyme which converts testosterone to estradiol-17beta, in the ovarian follicle of medaka (Oryzias latipes) is regulated at the transcriptional level. In this study, we cloned a cDNA encoding a FTZ-F1-like protein (mdFtz-F1) from ovarian follicles of medaka. In vitro translated mdFTZ-F1, and nuclear extract from medaka ovarian follicles, formed complexes with oligonucleotide probes containing putative orphan nuclear receptor binding motifs, which are present in the promoter region of the medaka P-450arom gene. The expression pattern of mdFtz-F1 transcripts during oogenesis coincides with that of P-450arom transcripts. Transfection assays further suggest a potential transcriptional regulatory activity of mdFTZ-F1 upon the medaka P-450arom promoter. Taken together, these results suggest a potential role of mdFTZ-F1 in the transcriptional regulation of P-450arom in the ovarian follicle of medaka. PMID- 10375034 TI - Long-term results of pediatric primary one-stage cholesteatoma surgery. AB - The long-term results of surgical treatment for pediatric cholesteatoma are variable and there is no consensus on operation methods and on factors affecting outcome of surgery. We analyzed the independently evaluated long-term results and possible reasons for recurrence of cholesteatoma. A total of 84 consecutive pediatric (age < 16 years) cholesteatoma operations in the Helsinki University Central Hospital ENT Department. The operations were not staged, and all mastoids were obliterated and bony ear canals reconstructed without open cavities. The pre and postoperative and annual control data were recorded in a database. The last control was independently performed (JS) with an average follow-up of 4.8 years and 87% attendance. The total recurrence rate was 29% (24/84), and it was not dependent on the size of cholesteatoma, mastoid status, cholesteatoma in the window niches or stapedial erosion. A retraction process developed in 25% (21/84) of the ears and 42% (9/21) of these turned into retraction pocket cholesteatomas as late as 13 years postoperatively. Retractions and postoperative discharge, especially in combination, predisposed to recurrence. Of the healed ears, 37% became atelectatic. Hearing was maintained on the preoperative level. Reduced middle ear and attic ventilation led to retractions, and atelectasis and a tendency to discharge accelerated the process. Pitfalls in mastoid obliteration and attic reconstruction and the failure to create new ventilation routes were important reasons for recurrence of cholesteatoma. PMID- 10375036 TI - Tympanometry by general practitioners: reliable? AB - BACKGROUND: The diagnosis of otitis media with effusion (OME) is difficult using only medical history and otoscopy. Tympanometry may, therefore, be helpful in the diagnosis and follow-up of OME in general practice. Studies regarding the reliability of tympanogram production and validation of tympanogram outcome have been performed. OBJECTIVE: To gain insight into the usability of microtympanometry and the degree of agreement and accuracy of tympanogram classification in general practice. METHODS: Data were collected in the offices of 49 general practitioners (GP's). The usability of the microtymp was monitored against a checklist. GP's (39) classified 47 tympanograms according to Jerger's modified classification, designating them as 'OME', 'no OME' or 'interpretion impossible'. The gold standard was the consensus over the 47 tympanograms reached by three doctors very experienced in tympanometry. RESULTS: Of the general practitioners, 61% handled the microtymp faultlessly. The overall inter-observer agreement was moderate to substantial; with respect to the gold standard 74% of the general practitioners had a satisfactory to almost perfect agreement. These results were achieved after instruction and training; longer practice produced no significant improvement in the agreement. CONCLUSION: After training and instruction microtympanometry is a reliable diagnostic instrument in general practice. The classification of tympanograms is satisfactory. Classification problems arise when the curve is not a good one. Additional criteria for the assessment of the curves are proposed. PMID- 10375035 TI - Adenotonsillectomy in the very young patient: cost analysis of two methods of postoperative care. AB - Postoperative management of the patient younger than 36 months undergoing adenotonsillectomy has been the subject of many debates. Concerns for early postoperative complications such as airway obstruction, emesis, dehydration, and hemorrhage have led many physicians to consider overnight hospitalization following adenotonsillectomy in very young children. Trends in health care management have had increasing focus on cost effective means of treating patients to limit unnecessary expenditure on the part of the patient, physician, and hospital facility. The purpose of this retrospective review was to analyze two methods of early postoperative management in children less than 36 months old undergoing adenotonsillectomy at the Children's Hospital, San Diego from 1992 to 1997. Three hundred and seven cases were reviewed. Same-day discharge was compared with overnight inpatient observation based on the cost analysis of these two methods of postoperative care. Postoperative care was based on length of stay in the recovery room and as an inpatient. Expense of postoperative care was based on cost calculation for the recovery room and overnight hospitalization. Of the 307 patients, 194 went home the day of surgery and 113 were observed overnight in the hospital. Average hospital cost was higher in the outpatient group than in the inpatient group (P < 0.001). This difference reflects longer recovery room stay (350 min) in the outpatient group compared to the inpatient group (108 min) (P < 0.001). Outpatient adenotonsillectomy in the patient under 36 months may be safe; however, prolonged recovery room stays may actually make outpatient surgery less cost-effective than overnight admission. Recovery room costs are significantly higher per unit time than costs of inpatient hospitalization. Further investigation of cost-effective outpatient observation units may improve cost containment in the outpatient surgical setting. PMID- 10375037 TI - Evaluation of the use of a questionnaire to detect hearing loss in babies in China. AB - A questionnaire was used to screen hearing of 1020 babies 6-8 months in China. All babies failing the questionnaire and 10% of those who passed were tested using auditory brainstem audiometry (ABR). Babies with unilateral or bilateral hearing thresholds 30 dBnHL or more were investigated to determine the cause of the hearing impairment. Sixty-seven failing the questionnaire were tested and 23 were confirmed to have a hearing loss, 20 with bilateral hearing impairment. The causes were: 13 otitis media with effusion (OME), one hypoxia, one genetic and five unknown. One child with an OME related hearing loss passed the screen. The sensitivity of the questionnaire was estimated to be 70%, specificity 96%. PMID- 10375038 TI - Turban pin aspiration; a potential risk for young Islamic girls. AB - A turban is a kind of headcover, worn for religious intentions. In Islamic countries, girls start to wear a turban with the onset of puberty. Turban pins (headscarf needles) are used for attaching the layers of turban to each other in order to keep it in a steady position around the head. Aspiration of these pins is investigated in accordance with age groups, pin characteristics and treatment. From 1987 through 1998, 63 girls were admitted to our department with turban pin aspiration. All patients were healthy prior to aspiration. The median age was 14 years. Foreign bodies were removed, either by rigid bronchoscopy (n = 57), flexible bronchoscopy (n = 2), laryngoscopy (n = 3) or thoracotomy (n = 1). Repeated bronchoscopy rate was 8% (n = 5) and we had no mortality. This recently recognized aspiration hazard can be minimized by using adhesive bands or snap fasteners, instead of pins, when wearing a turban. PMID- 10375039 TI - Predictive value of acoustic reflectometry (angle and reflectivity) and tympanometry. AB - OBJECTIVE: Tympanometry and acoustic reflectometry are suggested tools for confirmation of otoscopic diagnosis of secretory otitis media. The issues on sensitivity and specificity of both devices are contradictory. In this study, our purpose was to compare sensitivity and specificity of both devices and to look for whether it is possible to reach higher values by combining them. METHODS: This study included 150 normal ears and 150 ears with chronic effusion. In tympanometry, only B tracings were accepted as predictor of effusion. In acoustic reflectometry, reflectivity (cut point: 5) and curve angle with two cut-points (75 degrees and 90 degrees) were used. RESULTS: Acoustic reflectometry presented higher specificity by both reflectivity (cut point: 5) and by curve angle (cut point: 75 degrees) (99.33% by both) than tympanometry (92%) (chi2 analysis, P < 0.001). But, their sensitivities (65.33 and 78%) were lower than tympanometry (96%) (chi2 analysis, P < 0.001). With curve angle of 90 degrees, specificity of acoustic reflectometry decreased to 85.33%, sensitivity increasing to 97.33%, which was not different from tympanometry (chi2 analysis, P > 0.1). When data of curve angle and tympanometry were combined, specificity and sensitivity of the combined test were found to be 91.33 and 100%, respectively. CONCLUSIONS: (i) Acoustic reflectometry should not be proposed as a better device than tympanometry, because its test efficiency was not higher than tympanometry. (ii) The only advantage of AR (reflectivity > or = 5 and curve angle < or = 75 degrees) was its high specificity to effusion. In addition, higher curve angles than 90 degrees were found to be highly predictive for normal ears as much as tympanometry. But, predictivity of curve angle between 76 degrees and 90 degrees was low. (iii) When tympanograms and curve angle were combined, it was found that prediction of this combination for curve angles between 76 degrees and 90 degrees was perfect. (iv) We consider that both test devices provide complementary data to each other, which would be particularly important for screening studies and that they are good tools for confirmation of clinical impression, particularly for less experienced clinicians. PMID- 10375040 TI - Spread of amniotic fluid cellular content within the neonate middle ear. AB - Six full-term neonatal temporal bones, with meconium contaminated amniotic fluid aspiration of varying degrees, were serially sectioned at 20 microm and every tenth section was stained by hematoxylin eosin and mounted on slides. All stained sections were studied, the data recorded and relevant details of all compartments photographed. In addition, four normal neonate temporal bones were studied, one by serial sectioning and three by microdissection. The lateral incudomalleal and tensor folds were present in all, membrane defects in the tensor fold were seen in two normal ears. Three ears in the aspiration group had much fetal tissue present rendering Prussak's space small. Four ears with aspiration had remnants of incus intercrural (medial) folds. The amniotic fluid cellular content (AFCC) was sparse or nonexistent in the supratubal recess, Prussak's space and hypotympanum. It concentrated to the areas around the stapes, to sinus tympani and round window niche, to lower lateral attic and posterior pouch, medial attic and in lesser amounts to superior attic, mastoid antrum and air cells. Massive amounts of AFCC in tympanic isthmus and posterior pouch may lead to formation of granulation tissue and blockage of the aeration pathways to attic proper and to Prussak's space. These changes may initiate the development of chronic secretory otitis media in infants. PMID- 10375041 TI - The use of standardized orbital ultrasound in the diagnosis of sinus induced infections of the orbit in children: a preliminary report. AB - Infections of the orbit in children usually present as a complication of sinusitis and may result in blindness or even death. Orbital cellulitis (OC) and subperiosteal abscess (SPA) represent different pathologies within the spectrum of orbital infections. The differentiation between OC and SPA is important, since it implies two different therapeutic modalities. While SPA is usually treated by incision and drainage and parenteral antibiotics, OC may be treated with antibiotics alone. Contrast enhanced CT scan is commonly used in the diagnosis of orbital infections, but does not always prove accurate in differentiating between these two conditions. MRI is superior to CT in the resolution of soft tissue pathology and may be more precise in such situations, but is less available imaging tool outside North America and Europe. There have been a few reports in the early 1980's on the use of standardized orbital ultrasound (SOU) in these two conditions. We have used SOU in seven children with sinus induced orbital infections--four with SPA and three with OC. We reviewed our experience in these patients and compared the imaging characteristics of OC and SPA on SOU with those of conventional imaging modalities, used in orbital infections. In four of the cases, CT scan was inconclusive, while SUO was diagnostic. In this preliminary report, we conclude that SOU may be useful in the diagnosis of orbital infections. PMID- 10375042 TI - Bacteriology of the middle meatus in children. AB - Little is known about the bacteriology of the middle meatus in children. Therefore, middle meatal samples were obtained from 50 children who underwent adenoidectomy or adenotonsillectomy, while a group of 50 children submitted to minor non-ENT surgical procedures, were used as a control group. Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pneumoniae were the most frequent cultured organisms, not only in the ENT group (in 68, 50 and 60% of the children respectively) but also in the control group (40, 34 and 50%). These three potential pathogens were more frequently seen among the children of the ENT group but only for H. influenzae was the observed difference statistically significant (P = 0.009). On semiquantitative analysis, there seemed to be more negative cultures or cultures with only a few colonies in the control group, while the richer cultures were obtained from the ENT group. Again, only for H. influenzae, these differences reached a statistical significance (P = 0.003). Streptococcus viridans and Neisseria species, both organisms that might be able to inhibit colonisation by some of the pathogens, were more frequently cultured in the control than in the ENT group: Strep. viridans 30 vs. 10% (P = 0.025) and Neisseria species 14 vs. 2% (P = 0.069). PMID- 10375043 TI - Jugular bulb dehiscence in achondroplasia. AB - Jugular bulb dehiscence--complete absence of a roof over the jugular bulb--is a rare malformation, probably present in <<< 1% of the general pediatric population. Of 126 children with achondroplasia evaluated in the Midwest Regional Bone Dysplasia Clinic, four and probably five, were identified as having such dehiscence (at least 3.2% of the children assessed). Identifying this increased incidence in achondroplasia is of some clinical relevance, particularly including risk of difficult to control bleeding at myringotomy. It may also present as otherwise unexplained hearing loss, tinnitus and self audible bruits in these children. PMID- 10375044 TI - Conductive hearing loss in Beckwith-Wiedemann syndrome. AB - Beckwith-Wiedemann syndrome is a rare genetic overgrowth syndrome presenting with organomegaly, abdominal wall defects, macroglossia, and postnatal hypoglycemia. Head and neck manifestations of this abnormality include flame nevus of the forehead and characteristic sulci of the ear lobe. We present a 7-year-old child with Beckwith-Wiedemann syndrome and a rare finding of conductive hearing loss on both sides due to congenital malleus and stapedial fixation. Small fenestra stapedotomy and mobilization of malleus fixation in the epitympanum improved the child's hearing. The bony fixation of the malleus and stapes is explained as atavism of the processus anterior mallei and peripheral lamina stapedialis in embryological development. PMID- 10375045 TI - Insulin resistance in obese hypertensive Asian Indian subjects. PMID- 10375046 TI - Genetic and environmental influences on waist-to-hip ratio and waist circumference in an older Swedish twin population. AB - OBJECTIVE: To investigate the genetic and environmental influences on waist-to hip ratio (WHR) and waist circumference (WC) measurements in males and females. DESIGN: Measurements taken from 1989-1991 as part of The Swedish Adoption/Twin Study of Aging (SATSA) were used for analysis. The SATSA sample contains both twins reared together as well as twins reared apart. SUBJECTS: 322 pairs of twins (50 identical, 82 fraternal male pairs and 67 identical, 123 fraternal female pairs); age range: 45-85y (average age, 65y). MEASUREMENTS: Waist-to-hip ratio (WHR), waist circumference (WC) and body mass index (BMI). RESULTS: In males, additive genetic effects were found to account for 28% of the variance in WHR and 46% of the variance in WC. In females, additive genetic effects were found to account for 48% of the variance in WHR and 66% of the variance in WC. The remaining variance in males was attributed to unique environmental effects (WHR, 72%; WC, 54%) and in females the remaining variance was attributed to unique environmental effects (WHR, 46%; WC, 34%) and age (WHR, 6%). When BMI was added into these models it accounted for a portion of the genetic and environmental variance in WHR, and over half of the genetic and environmental variance in WC. CONCLUSION: There are both genetic and environmental influences on WHR and WC, independent of BMI in both males and females, and the differences between the sexes are significantly different. PMID- 10375047 TI - Sociodemographic distribution of measures of body fatness among children and adolescents in New South Wales, Australia. AB - BACKGROUND: Obesity in childhood and adolescence is associated with health problems, increases in cardiovascular disease (CVD) risk factors and a greater likelihood of becoming overweight as an adult. A description of the sociodemographic distribution of overweight and obesity among children and adolescents in the population may allow us to determine if health promotion resources should be differentially allocated to particular groups. METHODS: The New South Wales Schools Fitness and Physical Activity Survey, 1997 (n = 5518) was a cross-sectional survey which measured height, weight, waist and hip girths and skinfold thicknesses of randomly selected New South Wales students in school years 4, 6, 8 and 10. Height and weight only were measured among Year 2 students. The mean ages of students in school years 2, 4, 6, 8 and 10 were 7.3y, 9.3y, 11.3y, 13.3y and 15.3y, respectively. The values of body mass index (BMI), waist girth, waist-to-hip ratio (WHR) and sum of three skinfolds for students resident in urban and rural areas and in tertiles of socioeconomic status (SES) were compared. RESULTS: There were no differences on any of the anthropometric measures between urban and rural boys and girls, with the exception that WHR was higher among urban girls. Among boys, there were no differences between the SES tertiles on any of the measures. Among girls, each of the anthropometric measures (except sum of skinfolds) was inversely associated with SES, with body fatness tending to be lower in the high SES tertile, compared with the low and middle SES tertiles. None of the statistical interactions between school year and body fatness was significant, suggesting that the relationship is consistent from childhood to adolescence. CONCLUSION: On the basis of these results, we suggest that health promotion programs aimed at weight management among Australian girls of lower SES are not warranted. We recommend that health promotion programs emphasize regular physical activity, a healthy diet and acceptance of the normal range of body shapes. PMID- 10375048 TI - A peptide leptin antagonist reduces food intake in rodents. AB - OBJECTIVE: The purpose of the present study was to investigate the continuing validity of the hypothesis that leptin is a physiologically important regulator of food intake, using the human leptin mutant R128Q leptin. DESIGN: In a cellular proliferation assay, based on BAF-3 cells transfected with the murine ObRb receptor, R128Q leptin was shown to be devoid of agonistic activity and to competitively inhibit the proliferative effects of leptin. To determine whether R128Q leptin was also an antagonist of leptin in vivo, the leptin mutant was injected intracerebroventricularly (i.c.v.) into rats in the absence and presence of leptin. R128Q was also injected intraperitoneally (i.p.) into ob/ob and into db/db mice expressing, respectively, either normal or defective ObRb receptors. RESULTS: R128Q was shown to be a competitive antagonist of leptin induced cellular proliferation in vitro. Surprisingly, in vivo R128Q leptin produced a strong dose-dependent decrease in food intake, and was only slightly less potent than leptin itself. In fasted rats, the inhibitory effects of leptin and R128Q leptin (i.c.v.) on post-fast refeeding were additive. Finally, R128Q leptin produced the same inhibition of food intake as leptin when injected i.p. in ob/ob mice and, like leptin, was inactive after i.p. injection to db/db mice. CONCLUSION: R128Q leptin is a leptin agonist in vivo, but behaves as an antagonist against leptin induced proliferation in vitro. The data demonstrate that the human leptin mutant R128Q leptin is not a suitable tool for investigating the physiological actions of leptin. PMID- 10375049 TI - Leptin plasma levels as a marker of sparing-energy mechanisms in obese women. AB - OBJECTIVE: To investigate possible relationships between leptin and energy expenditure (EE), both in the condition of stable body weight and during weight loss. SUBJECTS: Seventy four Caucasian, adult obese women with stable body weight (including 10 obese women studied before and during a body weight-reducing program). MEASUREMENTS: Resting EE (REE) and substrate oxidation rates by indirect calorimetry; plasma leptin concentrations by radioimmunoassay (RIA). RESULTS: In conditions of stable body weight, leptin values showed a significant, negative relationship with REE, as expressed in absolute values (P = 0.030) and as adjusted for the variation in lean body mass (LBM) (P = 0.017). This negative relationship was independent of both LBM and fat mass (FM). Linear regression analysis was used to obtain the equation linking REE and LBM; then both predicted REE and the percent deviation from predicted REE were calculated for each subject. Leptin values were negatively related (P < 0.0001) to the deviation from predicted REE. During active body weight loss, the modifications of both REE (delta REE) and lipid oxidation (delta lipid oxidation) were significantly negatively related to leptin concentrations, which were measured before the dieting period (P < 0.03 for both). CONCLUSION: In obese women, high plasma leptin concentrations are associated with a low rate of REE, when body weight is stable, and with a reduction of REE and lipid oxidation, in response to a hypocaloric diet. This suggests that, in severely obese women, leptin is a marker of sparing energy mechanisms operating in both basal and reducing weight conditions. PMID- 10375050 TI - Physique, subcutaneous fat, adipose tissue distribution, and risk factors in the Quebec Family Study. AB - OBJECTIVE: To investigate the relationships among subcutaneous fatness, subcutaneous adipose tissue (SAT) distribution, somatotype and risk factors for coronary heart disease (CHD). SUBJECTS: The sample included 1410 (715 male and 695 female) youths and adults from the Quebec Family Study. MEASUREMENTS: Six skinfolds and the dimensions necessary for the derivation of the Heath-Carter anthropometric somatotype (endomorphy, mesomorphy, ectomorphy) were measured. The six skinfolds were summed to provide an index of subcutaneous adiposity (SUM). In addition, the trunk-to-extremity skinfold ratio, adjusted for SUM using regression procedures (TER), and the first principal component (PC1) of skinfold residuals (also adjusted for SUM) were used to indicate SAT distribution, independent of the overall level of fatness. Risk factors for CHD included systolic and diastolic blood pressures, and fasting glycaemia, triglycerides (TGs), plasma cholesterol, high and low density lipoprotein (HDL-C and LDL-C) cholesterol, and the HDL-C/total cholesterol (CHOL) ratio. RESULTS: In general, SUM was positively correlated with endomorphy and mesomorphy, and negatively correlated with ectomorphy. On the other hand, SAT distribution was not associated with somatotype, except in females where TER and PC1 were negatively correlated with mesomorphy. Results of forward stepwise regression analyses to predict CHD risk factors, indicated that a significant proportion of the variance in the risk factors could be accounted for by SUM, SAT distribution and somatotype (up to 16%). SUM is the best predictor, entering the regressions first (most important) in six of 15 significant regressions in males and 14 of 16 significant regressions in females. Somatotype components enter as predictors 10 times in males, and six times in females. Similarly, TER and PC1 enter as predictors nine times in males and five times in females. CONCLUSIONS: Somatotype is related to SUM, while somatotype and SAT distribution are largely independent of one another. Furthermore, SUM, somatotype and SAT distribution are significant predictors of biological risk factors for CHD. PMID- 10375051 TI - Binge status as a predictor of weight loss treatment outcome. AB - OBJECTIVE: A widely held clinical belief is that individuals with binge eating problems fare poorly in weight loss programs. The empirical evidence regarding the prognostic significance of binge eating, however, is mixed. The goals of this study were to examine psychological and behavioral characteristics associated with binge eating and the prognostic significance of binge eating for short- and long-term weight loss in a large sample of women treated for obesity. DESIGN: The dataset used in the current study was a combined sample of women (n = 444) who participated in one of three behavioral weight loss research studies. MATERIALS AND METHOD: Measures of dieting and weight history were obtained at baseline. Body weight, the Binge Eating Scale (BES), a measure of perceived barriers to weight loss, the Beck Depression Inventory, the Block Food Frequency Questionnaire, and the Paffenbarger Physical Activity Questionnaire were assessed at baseline, 6 months and 18 months. Regression analyses examined cross-sectional associations between the BES and the other variables at baseline, prospective associations between baseline BES and changes in weight and the psychological and behavioral variables over time, and temporal covariations between BES and the other variables over time. RESULTS: Cross-sectional analyses showed baseline binge eating status to be strongly associated with dieting history, weight cycling, depressive symptomatology and perceived barriers to weight loss. Women with binge eating problems were also more likely to drop out of treatment. Baseline binge status was not associated with 6-month weight loss, but was weakly predictive of less weight loss success at 18 months. Binge status at baseline did not predict changes in dietary intake, physical activity, perceived barriers to weight loss or depressive symptomatology at either 6 months or 18 months. In time dependent covariance analyses, changes in BES scores were significantly associated with changes in body weight, independent of changes in dietary intake and physical activity. However, when depression scores are included in the analysis, the association between binge score and body weight was no longer statistically significant. CONCLUSION: These findings suggest that baseline binge status was a weak prognostic indicator of success in women who are moderately obese and are seeking treatment for weight loss. Although assessments of binge status covary with weight loss and regain, the relationship appears to be mediated by psychological dysphoria. PMID- 10375052 TI - Effects of dietary restriction on serum leptin concentration in obese women. AB - OBJECTIVE: To investigate the short- and long-term effects of dietary restriction on serum leptin in obese women and the role of the gastrointestinal system in the short-term regulation of leptin production. DESIGN: Clinical longitudinal study of anthropometric and serum leptin changes induced in obese women by a balanced 300 kcal/d very low calorie diet (VLCD), administered either orally or parenterally for 5 d, and by a balanced 900 kcal/d low calorie diet (LCD) lasting six months. SUBJECTS: 20 obese women (age: 38.1 +/- 12.7 y; body mass index (BMI): 40.2 +/- 8.3 kg/m2). RESULTS: Five days following VLCD, a modest, even if significant (P < 0.0001), fall of both body weight (BW) and BMI was observed, along with a dramatic (> 50%) highly significant (P < 0.0001) reduction of circulating serum leptin. Baseline and five-day anthropometric and biochemical findings were closely similar in the group of orally fed subjects, when compared with those of their parenterally fed counterparts. The baseline positive correlation between serum leptin and BMI (p = 0.533) increased (P < 0.05) at the end of the five day VLCD (p = 0.849). A further fall of BW and BMI was observed at day 30 (P < 0.001) and day 180 (P < 0.01) during the 900 kcal/d LCD, while the serum leptin concentration gradually increased until day 180 when it was only slightly but non significantly lower than at baseline. At the end of the study, the correlation between serum leptin and BMI was similar to the baseline (p = 0.562). CONCLUSIONS: Energy restriction causes a fall of serum leptin apparently not mediated by gastrointestinal signals and it seems not to affect the long-term regulatory pathways of circulating leptin. PMID- 10375053 TI - Association of waist circumference with ApoB to ApoAI ratio in black and white Americans. AB - BACKGROUND: Although numerous studies have demonstrated obesity as an aspect of the insulin resistance syndrome in cardiovascular disease (CVD), the mechanism is not clear. Central adiposity, acting through many CVD risk factors, including, plasma glucose, insulin, total cholesterol, low density lipoprotein-cholesterol (LDL-C) and lipoprotein moities-apolipoprotein B (ApoB), apolipoprotein A-I (ApoAI), by atherogenic and thrombotic mechanisms has been proposed as a possible mechanism. In this study, we examined the relationship between central fat distribution (defined by waist circumference) and the ratio of these lipoproteins (ApoB/ApoAI). SUBJECTS AND METHODS: Association between ApoB/ApoAI ratio and waist circumference was compared in Blacks (n = 854) and Whites (n = 2552) using the NHANES III population-based samples. Correlation analyses and multiple regression analyses were used to determine the association between ApoB/ApoAI and waist circumference, controlling for age, body mass index (BMI), race, gender, plasma glucose, insulin, serum triglyceride and total cholesterol. RESULTS: Adjusting for age, ApoB/ApoAI was significantly correlated with waist circumference (Black men: r = 0.38, White men: r = 0.26, Black women: r = 0.20, White women: r = 0.36) (all P < 0.01). Adjusting for age and triglyceride or insulin, waist circumference was also positively correlated with CVD risk factors including, ApoB, LDL-C, plasma glucose and fasting insulin, and inversely correlated with ApoAI and HDL-C in Blacks and Whites (P < 0.05). Overall, triglyceride and total cholesterol were the strongest predictors of ApoB/ApoAI in Blacks and Whites adjusting for age, BMI and insulin, than waist girth (P < 0.01). CONCLUSIONS: The result of this study suggests the need to investigate ApoB/ApoAI as another possible facet in the insulin resistant syndrome. PMID- 10375054 TI - The Obesity Adjustment Survey: development of a scale to assess psychological adjustment to morbid obesity. AB - OBJECTIVE: To develop a reliable and valid measure of distress, related to extreme obesity. DESIGN: Items related to distress over obesity were selected from the literature, clinical experience and from input provided by a gastroplasty patient support group. The items were assessed in a longitudinal study, with the body mass index (BMI) and psychological assessment occurring 2-6 months prior to, and 12 months following, gastroplasty surgery. SUBJECTS: 81 females and eight males (mean age 35.9 y) who had been accepted for gastroplasty surgery. All but two of the patients had BMIs > 40 (Mean = 48.11, s.d. = 6.84). MEASUREMENTS: BMIs were calculated using weight and height. Psychological characteristics were assessed using the Mental Health Inventory (MHI), the Sickness Impact Profile (SIP), and the Eating Inventory (EI). Demographic information was collected with a questionnaire. RESULTS: Attempts to factor analyse the 95 item questionnaire were unsuccessful. Alternatively, a shorter, 20 item questionnaire was developed. The questionnaire shows good test-retest reliability (r = 0.867), good internal consistency (coefficient alpha = 0.719), good face and construct validity, and is sensitive to pre-post surgical change. CONCLUSIONS: The Obesity Adjustment Survey (OAS) may be useful as a brief measure of distress in obese individuals. This measure can be used to index the psychological impact of gastroplasty surgery on psychological functioning, and can be used in future research as a disease-specific measure to predict success of surgery. PMID- 10375055 TI - Assessment of insulin sensitivity from plasma insulin and glucose in the fasting or post oral glucose-load state. AB - OBJECTIVE: To compare insulin sensitivity indexes derived from plasma insulin (I) and glucose (G) in the basal state (Sib) and at the second hour (I2h and G2h) of an oral glucose tolerance test (OGTT, Si2h) (i) with measurements of insulin sensitivity using the insulin modified frequently sampled intravenous glucose tolerance test (FSIVGTT) [Si(IVGTT)] and (ii) with modelling of fasting glucose and insulin by the homeostasis model assessment (HOMA). SUBJECTS: 47 subjects entered the study. 31 subjects were classified as having normal glucose tolerance (NGT), 10 as having impaired tolerance to glucose (IGT) and six as type 2 diabetes mellitus according to the World Health Organisation (WHO) criteria. MEASUREMENTS: Sib and Si2h were calculated as follows. Sib = 10(8)/(I x G x VD), Si2h = 10(8)/(I2hr x G2hr x VD) where VD is an estimate of the apparent glucose distribution volume. A third insulin sensitivity index (SiM) was calculated by averaging Sib and Si2h. HOMA was calculated as follows: I/(22.5 x e(-lnG)) RESULTS: Si(IVGTT), Sib, SI2h and SiM were all significantly higher in subjects with NGT than in those with IGT or type 2 diabetes. Si(IVGTT) was highly correlated (P < or = 0.0001) with the three insulin sensitivity indexes found in the total population, in subjects with NGT and in those with IGT. In type 2 diabetic patients, a significant correlation was only noted when SiM was tested against Si(IVGTT) (P < or = 0.05). In most circumstances, the associations of Si(IVGTT) with Sib, SI2h and SiM were stronger than the corresponding associations with Ib, I2h or HOMA. SiM was the index that correlated best with Si(IVGTT) in the whole group (r = 0.92, P < 0.0001) as well as in NGT (r = 0.86, P < 0.0001), IGT (r = 0.96; P < 0.0001) and type 2 diabetes (r = 0.83, P < or = 0.05) subgroups. CONCLUSIONS: Calculations of sensitivity indexes from G and I concentrations in the basal state and during a conventional 2 h OGTT appear to be useful for coupling in the same simple and single test both a determination of glucose tolerance and an estimate of insulin sensitivity. PMID- 10375056 TI - Effects of hormone replacement therapy and social stress on body fat distribution in surgically postmenopausal monkeys. AB - OBJECTIVE: To investigate the effects of hormone replacement therapy (HRT) and social stress on body fat distribution in an animal model of women's health, the female cynomolgus macaque (Macaca fascicularis). DESIGN/SUBJECTS: Adult female cynomolgus monkeys were ovariectomized and fed an atherogenic diet for two years while housed in social groups of 3-8 monkeys each. Animals were then fed a lipid lowering diet and randomized into four experimental groups: a baseline group which was necropsied immediately and not included in the study reported here, 26 females fed diet only (CONTROL), 22 females fed diet plus conjugated equine estrogens (CEE), and 21 females fed the diet plus CEE and medroxyprogesterone acetate (CEE + MPA). Treatment lasted 30 months. MEASUREMENTS: During the last nine months of treatment, social status was determined three times at three month intervals. At the end of the study, whole body obesity and fat distribution patterns were determined using anthropometry and computerized tomography (CT). RESULTS: The addition of a progestin to the estrogen replacement regimen administered to surgically postmenopausal monkeys, increased all anthropometric and CT measures of obesity except intra-abdominal fat. HRT had no effect on patterns of fat distribution. Socially-dominant, ovariectomized females were more obese than subordinates using both anthropometric and CT measurements of whole body obesity. Dominant females were more likely to have their fat deposited centrally as measured anthropometrically. However, CT measures revealed a trend for dominants to preferentially deposit fat in the subcutaneous abdominal depot in contrast to subordinates who deposited fat in the intra-abdominal depot. CONCLUSIONS: The results of this study suggest that progestins, when administered in combination with estrogens, may increase fat deposition, particularly in subcutaneous depots. In addition, the social stress experienced by subordinate monkeys, may have mild effects on fat deposition patterns, even after removal of ovarian function as a factor. These observations may have implications for treatment recommendations in postmenopausal women. Lastly, CT may measure different characteristics of fat distribution than skinfolds and circumferences. PMID- 10375057 TI - Randomized trial on protein vs carbohydrate in ad libitum fat reduced diet for the treatment of obesity. AB - OBJECTIVE: To study the effect on weight loss in obese subjects by replacement of carbohydrate by protein in ad libitum consumed fat-reduced diets. DESIGN: Randomized dietary intervention study over six months comparing two ad libitum fat reduced diets (30% of total energy) strictly controlled in composition: High carbohydrate (HC, protein 12% of total energy) or high-protein (HP, protein 25% of total energy). SETTING AND PARTICIPANTS: Subjects were 65 healthy, overweight and obese subjects (50 women, 15 men, aged 18-55 y) randomly assigned to HC (n = 25), HP (n = 25) or a control group (C, n = 15). All food was provided by self selection in a shop at the department, and compliance to the diet composition was evaluated by urinary nitrogen excretion. MAIN OUTCOME MEASURE: Change in body weight, body composition and blood lipids. RESULTS: More than 90% completed the trial. Weight loss after six months was 5.1 kg in the HC group and 8.9 kg in the HP group (difference 3.7 kg, 95% confidence interval (CI)(1.3-6.2 kg) P < 0.001), and fat loss was 4.3 kg and 7.6 kg, respectively (difference 3.3 kg (1.1-5.5 kg) P < 0.0001), whereas no changes occurred in the control group. More subjects lost > 10 kg in the HP group (35%) than in the HC group (9%). The HP diet only decreased fasting plasma triglycerides and free fatty acids significantly. CONCLUSIONS: Replacement of some dietary carbohydrate by protein in an ad libitum fat-reduced diet, improves weight loss and increases the proportion of subjects achieving a clinically relevant weight loss. More freedom to choose between protein-rich and complex carbohydrate-rich foods may allow obese subjects to choose more lean meat and dairy products, and hence improve adherence to low-fat diets in weight reduction programs. PMID- 10375058 TI - The impact of body build on the relationship between body mass index and percent body fat. AB - OBJECTIVE: The objective of the study was to test the hypothesis that differences in the relationship between percent body fat (%BF) and body mass index (BMI) between populations can be explained (in part) by differences in body build. DESIGN: Cross-sectional, comparative study. SUBJECTS: 120 age, gender and BMI matched Singapore Chinese, Beijing Chinese and Dutch (Wageningen) Caucasians. MEASUREMENTS: From body weight and body height, BMI was calculated. Relative sitting height (sitting height/height) was used as a measure of relative leg length. Body fat was determined using densitometry (underwater weighing) in Beijing and Wageningen and using a three-compartment model based on densitometry and hydrometry in Singapore. Wrist and knee widths were measured as indicators for frame size and skeletal mass was calculated based on height, wrist and knee width. In addition, a slenderness index (height/sum of wrist and knee width) was calculated. RESULTS: For the same BMI, Singapore Chinese had the highest %BF followed by Beijing Chinese and the Dutch Caucasians. Singaporean Chinese had a more slender frame than Beijing Chinese and Dutch Caucasians. Predicted %BF from BMI, using a Caucasian prediction formula, was not different from measured %BF in Wageningen and in Beijing, but in Singapore the formula underpredicted %BF by 4.0 +/- 0.8% (mean +/- s.e.m.) compared to Wageningen. The difference between measured and predicted %BF (bias) was related to the level of %BF and with measures of body build, especially slenderness. Correction for differences in %BF, slenderness and relative sitting height, decreased the differences between measured and predicted values compared to the Dutch group from 1.4 +/- 0.8 (not statistically significant, NS) to -0.2 +/- 0.5 (NS) in Beijing and from 4.0 +/- 0.8 (P < 0.05) to 0.3 +/- 0.5 (NS) in Singapore (all values mean +/- s.e.m.). CONCLUSIONS: The study results confirm the hypothesis that differences in body build are at least partly responsible for a different relationship between BMI and %BF among different (ethnic) groups. PMID- 10375060 TI - Cimetidine and obesity: conflicting evidence. PMID- 10375059 TI - Haemodynamic response to an isometric exercise test in obese patients: influence of autonomic dysfunction. AB - OBJECTIVE: To investigate blood pressure (BP) and heart rate (HR) responses to an isometric exercise test in obese non diabetic patients and to correlate the results with vagal function and plasma insulin concentration. SUBJECTS: 63 obese patients, 36 of whom had abnormal cardiac parasympathetic control (PS+), and 35 healthy control subjects. METHODS: Analysis of HR variations during three standardized tests: deep-breathing, lying-to-standing and Valsalva. Isometric contraction (handgrip) for 5 min. RESULTS: In the PS+ obese patients, resting HR and body mass index (BMI) were significantly higher than in the PS- subjects and there was a trend to higher plasma insulin concentrations. Age-matched comparison showed that during the handgrip test, the increase in HR at the first minute was significantly higher in the PS- obese patients than in the controls. The increase in BP was significantly lower in the PS+ obese patients than in age-and-BMI matched PS- obese patients. CONCLUSION: These data suggest that 1) there is an increase in cardiac vagal tone in PS- obese patients, since the early increase in HR at 1 min of the handgrip test, results from the withdrawal of vagal tone; 2) BP response to an isometric contraction is impaired in PS+ obese patients due to a lower sympathetic activation; 3) high plasma insulin concentrations may also contribute to limiting the BP response; and 4) autonomic disorders may account for alterations in the haemodynamic changes during exercise. PMID- 10375061 TI - Correlates of alcohol and drug use among low-income Hispanic immigrant childbearing women living in the USA. AB - Alcohol and drug use is a widespread and serious problem with deleterious consequences for the health and well-being of childbearing-age women and their children. Little information exists regarding etiological factors for substance use among Hispanic childbearing-age women immigrating to the United States (USA). This research provides a correlational analysis of factors associated with alcohol and drug use. The Social Stress Model for Substance Use Prevention provided the conceptual framework for this cross-sectional, interview administered survey of 60 low-income predominantly Mexican-American women. The outcome variable was alcohol and drug use (alcohol, cigarettes, marijuana, cocaine and opiates). Independent variables included the major constructs of the model: stress, social support, social influences, personal competencies and community resource utilization patterns. Findings suggested that the levels of drug use were lower among this study sample than in the general USA population regardless of pregnancy status. Bivariate correlations demonstrate that women with higher drug use indices had more lenient attitudes regarding drug use and were more likely to have family and friends that used alcohol and drugs. Although drug use was relatively low among this sample of women, both women who used alcohol themselves and women whose partners used alcohol and drugs reported significantly higher levels of stress, weaker social support and poorer levels of self esteem. Implications for practice and future research are suggested. PMID- 10375062 TI - Culture, conceptive technology, and nursing. AB - Technology is a form of cultural expression, formed of and forming culture. A paradox about technological innovation is that, in addition to creating new human arrangements and possibilities, it often serves only to reinforce existing sociocultural practices, norms, and values. The technologically radical is often the culturally conservative. Conceptive technology has contributed toward the redefinition of patienthood, the multiplication of models of infertility, and the reinforcement of existing cultural norms. Nurses are well-positioned to conduct a kind of technology assessment that places culture and ethics at the center of inquiry. They are also well-positioned to assist women and their partners seeking technological assistance to reproduce to understand the controversies concerning conceptive technology that may account for their own ambivalence toward continuing or terminating medical treatment, societal ambivalence toward supporting expensive fertility treatments, and cultural ambivalence toward technological development. PMID- 10375063 TI - Implementation of research findings to reduce postoperative pain at night. AB - This study was designed to introduce and evaluate a research-based intervention to improve night-time pain management. It involved the provision of patient information and the introduction of structured night-time pain assessment. The implementation of the intervention was undertaken by local opinion leader. The study involved 417 patients from two matched orthopaedic wards in a before and after trial with comparison group. Outcomes of night-time pain control were elicited from ward documentation and patients by structured interviews on the second postoperative morning. These incorporated retrospective pain assessments, analgesic provision and nursing comfort measures provided the previous night. The intervention was associated with statistically significant reductions in both average and worst overnight pain scores. The frequency of night-time pain assessment by nursing staff increased significantly, although patients did not volunteer reports of pain more frequently and analgesics and other comfort measures were no more frequent. The intervention required an investment in educational support but no additional resources were needed for the successful reduction in pain scores. PMID- 10375064 TI - The views of nurses to the conduct of a randomised controlled trial of problem drinkers in an accident and emergency department. AB - The Trent Regional Health Authority funded a study in 1995 to train nurses in an accident and emergency (A&E) department to screen all adult attendees for alcohol problems with a view to identifying a sample of problem drinkers to participate in a randomised controlled trial (RCT). In the RCT identified drinkers were to be assigned either to health education plus brief counselling intervention or, as controls, to health education alone. Despite 16654 attendance's at A&E during the recruitment phase of the study only 20% of attendees were screened of whom a further 19% were identified as problem drinkers by the CAGE screening questionnaire. Less than half of the problem drinkers were, however, provided with feedback by the nurses, leaving a small group of 264 eligible for entry to the RCT. The great majority of this subgroup refused an initial appointment at the specialist clinic and so the trial was abandoned. A number of in-depth interviews were undertaken with the nurses in an attempt to understand ways in which the overall conduct of the study might have been improved. This paper outlines in some detail some of the reasons for the lack of success with the study which include; general environmental factors that undoubtedly led to stress and poor morale amongst the nursing team, the differences in perception between managers and clinical nurses concerning the value of research and the inadequacy of the initial training programme. The paper concludes that there are problems in the NHS which do not provide a helpful backcloth to the successful conduct of health services research. PMID- 10375065 TI - How students experience professional socialisation. AB - The question of how a nurse becomes socialised into the nursing profession remains of critical importance. An exploration of the literature relating to professional socialisation reveals a shift from the notion that it is a reactive process, to proactive. Our research explores this issue from a personal constructivist perspective using the repertory grid technique. Our findings show that the professional socialisation process is complex and diverse. During their educational preparation community nursing students make a radical reappraisal of their role perceptions. In their transition to becoming a graduate practitioner they gain a greater understanding of their specialist role whilst becoming less rigid in their thinking. We conclude that the impact nurse education has on professional socialisation will depend on the students' past experiences, the reflective nature of the process and the beliefs and values promoted in the course. PMID- 10375067 TI - The nurse practitioner: redefining occupational boundaries? AB - This paper explores aspects of the controversy and conflict that has arisen within the nursing and medical professions regarding the emergence of nurse practitioners in the United Kingdom (U.K.). Difficulties in establishing satisfactory definitions of nurse practitioners, that allow them to be viewed decisively either within nursing or medical occupational roles, are discussed. The paper argues that the key to the debate may hinge on professional and occupational boundary redefinition which is currently resisted by some members of both the nursing and medical professions. The idea that nurse practitioners may be an evolving and discrete professional group, outside the currently accepted professional and occupational definitions of nursing and medicine, is explored. It is argued that both nursing and medicine are faced with a particular challenge in the nurse practitioner movement that is resulting in conflict as new boundaries are established. PMID- 10375066 TI - Knowledge, attitudes and beliefs about psychotropic medication among Saudi hospitalized psychiatric patients. AB - This quantitative descriptive study describes the knowledge, attitudes and beliefs about medication in a sample of 76 Saudi hospitalized psychiatric patients. Forty-four percent of the patients named their medication, 37% identified their side effects, and 49% knew the dosage. Chi-square results indicated that younger patients and college educated patients were significantly more knowledgeable about their medication doses and dose frequencies than older patients and less educated patients respectively. Shorter illness duration was associated with knowledge of side effects. Most patients thought they would stop their medications when they felt better and would not tell others they take psychotropic drugs. Study findings suggest the need for an active nursing role in psychoeducational interventions and further research. PMID- 10375068 TI - The effect of student nurses' experiences over the Common Foundation Programme on their inferences of suffering. AB - Poor pain assessment contributes to inadequate postoperative pain relief. Studies in the U.S.A. suggest that nurse education might make students less sensitive to patients' experience of pain. This research examined this process and the factors that influence it in the U.K. Two-hundred and seventeen students completed the Standard Measure of Inferences of Suffering Questionnaire (SMIS) before and after their Common Foundation Programme (CFP). Their inferences of psychological distress increased as studies in the U.S.A. had found but, unlike these studies, no change was found in their inferences of pain. These findings have important implications for both nurse education and the mechanisms to support student nurses in clinical practice. PMID- 10375069 TI - Factors associated with burden in primary caregivers of mentally ill patients. AB - The objectives of this study are to describe caregivers' subjective burden and to identify the predictors of burden in primary caregivers of mentally ill outpatients recruited from eight hospitals in Montreal, Quebec, Canada. Patient and primary caregiver variables, were regressed on perceived burden using hierarchical regression analysis. The variables describing the patient's current state contributed the most to explaining variance in subjective burden. The variables related to psychiatric history and to outpatient treatment also explained a significant proportion of the variance in the burden scores. Better understanding of the factors associated with subjective burden will enable researchers and practitioners to identify those caregivers that are at greater risk for higher levels of burden, and to develop more focused and appropriate interventions. PMID- 10375070 TI - Nurse education in an international context: the contribution of contingency. AB - Attempts to reform nurse education in the U.K. have met with limited success. A brief examination of similar moves in other countries reveals a similar situation. Placing experiences in this country in the context of global reform, it is possible to suggest that three sets of conditions need to be satisfied for change to follow: these relate to context, convergence and contingency. Context refers to the creation of a positive climate of opinion or a case and pressure for change. Convergence refers to the fortuitous fusion of professional and government agendas. Contingency provides the unforeseen consequence, the spark that ignites a policy change. The implications for further educational reform in this country are briefly discussed. PMID- 10375071 TI - The role of natural killer cell mediated caspases activation in a graft-versus host disease model of semiallogeneic small bowel transplantation. AB - In the clinical setting of solid organ transplantation the event of graft-versus host disease (GvHD) is rare and not easily predictable. Even intestinal and multivisceral transplants harbour a huge amount of immunocompetent cells and they do not exert a significantly higher risk to trigger serious GvH reactions. A series of our own experimental studies has been conducted to delineate the role of the host's innate immune system in the context of GvHD following parental to F1 hybrid semiallogeneic small bowel transplantation (SBTx). These results clearly demonstrated the immunological significance of the recipient's status of natural killer (NK) cell activity to counteract donor-derived lymphocytes and related cytotoxicity. NK cells and macrophages are both endowed with Ca2+ dependent receptors of the C-type lectin family which interact with a diversity of high-affinity oligosaccharide ligands expressed on potential target cells. One of these proteins of the C-type lectin family, termed NKR-P1, has been cloned and sequenced. Activation of NKR-P1 stimulates activation-induced cell death (AICD) of bound target cells. As intracellular mediators of apoptotic cell death a new family of cysteine proteases, the caspases, have been defined. These proteases appear to be involved in the initiation of apoptosis in response to a number of stimuli. This study was conducted to investigate the impact on the activity level of host NK cells and on target cell lysis of donor-derived lymphocytes after heterotopic semiallogeneic (parental [DA;RT1.aaav1] to F1 [DA x LEW;RT1.(1)]) small bowel transplantation using a rat model. The host's NK activity was either specifically activated (by use of polyinosinic:polycytodilic acid [poly-I:C]) or suppressed (by depletion of host NK cells after intraperitoneal administration of the NKR-P1 monoclonal antibody 3.2.3). The impact of NK-activity on the incidence of GvHD and the recipients' survival was correlated with the frequency of apoptotic cell death and related expression of caspases 1 (ICE) and 3 (CPP-32) from donor and recipient small bowel tissues. Our results confirm that depletion of NK cells in F1 host rats prior to parental small bowel transplantation significantly decreased the mean survival to 11.4 days versus 16.2 days of nondepleted F1 rats (p < 0.01). Conversely, activation of host NK activity with poly-I:C abrogated GvHD in all 12 recipient rats and led to long-term survival in seven of 12 animals. Long-term survival was associated with a substantially higher frequency of apoptotic cell death in donor and recipient small bowel and mesenteric lymph nodes. On day 10 after transplantation, Northern blot analysis of these tissues revealed profound upregulation of mRNA-specific gene expression for caspase 1 and 3 as potential mediators of programmed cell death of activated lymphocytes. Our findings emphasize the importance of NK cell associated innate immunity in the context of GvHD after semiallogeneic small bowel transplantation. Killing of alloreactive donor-derived lymphocytes was mediated by the NKR-P1 protein on NK cells and could be suppressed after pretreatment of F1 hosts with anti-NKR-P1 mAb 3.2.3. Moreover, NK cell-mediated apoptosis induced upregulation of caspases 1 and 3, thus elucidating the involvement of this protein in the context of caspase-mediated target cell killing. PMID- 10375072 TI - Effective chemokines and cytokines in the rejection of human retinal pigment epithelium (RPE) cell grafts. AB - OBJECTIVE: In the rejection of transplanted retinal pigment epithelium (RPE) cells, an activation of allografts is probably the pivotal point for long-term success. The detailed immunological interactions involved in the rejection after RPE transplantation are still unknown. The aim of this study is to evaluate the interactions of pro-inflammatory cytokines and chemokines in this activation process in vitro. METHODS: Human RPE cells (2 x 10(5)/ml) were therefore activated through a pre-treatment with different concentrations of interferon (IFN)-gamma (100 or 1000 U/ml), tumour necrosis factor (TNF)-alpha (1 or 10 ng/ml) or combinations of both, or employed in a nonactivated form. Afterwards, the RPE cells were tested by enzyme-linked immunosorbant assay (ELISA) and ribonuclease protection assays (RPA) for the secretion and mRNA content of the different chemokines (RANTES, MCP-1 and IL-8) and cytokines (IL-6) at various time points up to 48 h. MAIN FINDINGS: HRPE cells secrete the investigated cytokines in response to pro-inflammatory activation. This could be demonstrated at both the mRNA (RPA) and the protein levels (ELISA). The secretion was time and dose dependent, and significantly upregulated in comparison to that observed with nonactivated cells. CONCLUSIONS: This study demonstrates that RPE cells efficiently secrete such cytokines as RANTES, MCP-1, IL-6, and IL-8, and have an accountable neutrophil and monocyte chemotactic activity. Thus, it could be indicated that the investigated cytokines play a central role in the activation cascade of RPE and in RPE rejection as well. PMID- 10375073 TI - HLA-A and -B alleles in cornea donors as risk factors for graft rejection. AB - We determined the human leucocyte antigen (HLA)-A, -B and -DR allele frequencies in recipients and donors of 115 cornea transplants, for recipients who developed graft rejection and those who did not. No difference in HLA allele frequencies of the recipients was found. The frequencies of the HLA-A26, -B35 and -B44 alleles in cornea donors were increased in recipients who developed graft failure. The detrimental effect on corneal graft survival of these alleles was significant (p < 0.001). No such effect was observed in renal transplantation. Corneal graft survival was similar when one or two A26, B35 or B44 alleles were present on the donor cornea. The negative effect was similar in magnitude to the previously reported negative effect of an HLA-B locus match between donor and recipient. When both a B-locus match and an A26, B35 or B44 allele were present, the negative effect on graft survival was twice as strong, indicating that different immune mechanisms are responsible for these phenomena. PMID- 10375074 TI - Tolerance induction permits the development of graft-versus-host disease: donor mediated attack following small bowel transplantation in mixed chimeras. AB - The induction of tolerance to organ allografts would eliminate acute and chronic rejection as well as the need for nonspecific immunosuppression. We have shown that tolerance induced through the creation of mixed allogeneic bone marrow chimeras allows for the long-term engraftment of cardiac and small bowel allografts across strong multiple major histocompatibility barriers. The possibility that tolerance might render the host susceptible to graft-versus-host disease (GVHD) has not been investigated in this or other models of tolerance. To test this possibility chimeras were created by transplantation of T-cell depleted ACI and Lewis bone marrow into lethally irradiated Lewis rats. Chimerism was determined post bone marrow transplant (BMTx) by flow cytometry of lymphocytes from reconstituted animals. ACI/Lew chimeras (ALC), Lewis/ACI F1 (LACF1), and Lewis (LEW) rats all received heterotopic ACI vascularized small bowel grafts. A second group of chimeras received small bowel grafts from ACI rats pretreated with low dose irradiation to eliminate T-cells from the graft. LEW-->LEW small bowel isografts were also performed. Animals were examined for evidence of GVHD by clinical signs and histologic examination of biopsied tissues. GVHD was quantified using the popliteal lymph node enlargement assay. All LACF1 rats developed severe lethal GVHD following ACI small bowel transplant. Bone marrow chimeras, ALC (n = 6), developed fatal GVHD in a similar fashion after receiving a small bowel transplant. LEW-->LEW isografts and chimeras receiving bowel from irradiated ACI rats survived long term without GVHD while ACI-->LEW allogeneic transplants all underwent acute rejection. GVHD or its absence was confirmed histologically. Popliteal lymph node enlargement indices reflected the presence of GVHD in the chimeras (1.87) and LACF1 (5.4) receiving allografts, but not in isografts or chimeras receiving irradiated allogeneic transplants. Analysis of cytokines in the tongues of rats undergoing GVHD showed elaboration of Th1 type proinflammatory cytokines which was not seen in isografted rats or rats receiving preirradiated small bowel. These results demonstrate that tolerance induction through mixed chimerism results in susceptibility to small bowel induced GVHD. Preirradiating the donor bowel prior to SBTx can prevent GVHD. PMID- 10375075 TI - Apoptosis and expression of cytotoxic T lymphocyte effector molecules in renal allografts. AB - Cytotoxic T lymphocyte (CTL) mediated apoptosis is thought to play a major role in the rejection of renal allografts following transplantation, however, the CTL effector mechanism that is primarily responsible for immunological rejection is unknown. The two major effector pathways of CTL killing which lead to apoptosis involve the Fas/Fas ligand (Fas L) lytic pathway, and the perforin/granzyme degranulation pathway. The expression of CTL effector molecules which influence these pathways include Fas, Fas L and TiA-1 (cytotoxic granule protein). This study has investigated apoptosis by in situ terminal deoxytransferase-catalysed DNA nick end labelling (TUNEL), and the expression of CTL effector molecules by immunohistochemistry, in renal allograft biopsies obtained from patients following kidney transplantation. Renal biopsies were classified into three histological groups; acute cellular rejection, chronic rejection, or no rejection. The extent of T-cell infiltration of renal tissues was assessed by immunohistochemical staining with an anti-CD3 monoclonal antibody. Numerous TUNEL positive cells were detected in all transplant biopsies examined; these consisted mainly of renal tubular cells and infiltrating cells, with some TUNEL positive cells also detected in the glomeruli. In the case of normal kidney tissue, renal cells also stained positive for TUNEL but there was no lymphocytic infiltration. There was significantly more T-cell infiltration observed in acute rejection biopsies compared to the no rejection biopsies. In the case of Fas L expression, there was little expression in all three biopsy groups, apart from one case of chronic rejection. Conversely, although there were no significant differences in TiA-1 expression between the three biopsy groups, TiA-1 expression was more prominent in acute rejection biopsies. Furthermore, Fas expression was significantly decreased in acute rejection biopsies when compared to those of chronic and no rejection in which Fas was predominantly localized in the renal tubular cells. These results indicate that the mechanism of CTL killing leading to the rejection of renal allografts may be different in acute and chronic rejection. Moreover, our data indicate the potential for cytotoxic granule-based CTL killing in acute renal allograft rejection but not in chronic rejection. PMID- 10375076 TI - Assessment of peripheral tolerance in anti-CD4 treated C57BL/6 mouse heart transplants recipients. AB - The study was designed to compare second heart and skin grafts and in vitro assays as a means of assessing peripheral tolerance in C57BL/6 mice. Vascularized heterotopic BALB/c hearts were placed in C57BL/6 recipients treated with anti CD4, GK1.5 (1 mg total per 20 g mouse i.p. on days 0, 1, 2, 3). Those mice in which hearts survived for >60 days were challenged with donor and third-party (CBA) skin grafts or with second heart grafts, of donor or third-party origin, attached to the carotid artery and jugular vein. In vitro alloreactivity was assessed by mixed lymphocyte reactions (MLR) and cell mediated lympholysis (CML) using recipient spleen cells. Parenchymal damage, cellular infiltration and vascular disease were assessed from the histology of long-term allografts and isografts. Allografts in untreated recipients were rapidly rejected while isografts survived > 100 days. Primary allografts in anti-CD4 treated recipients also survived > 100 days, as did donor strain secondary heart transplants given at >60 days after the first graft. Third-party hearts were rapidly rejected, as were donor and third-party skin grafts placed on recipients with long-term allografts. These recipients showed low MLR response to both donor and third party stimulators and donor-specific suppression of CML at 60 days post graft. Long-surviving heart allografts all showed evidence of parenchymal damage and vascular intimal thickening. Thus in the BALB/c to C57BL/6 donor-recipient strain combination, hearts, but not skin grafts, could be used to demonstrate peripheral tolerance, which seemed to be both organ and major histocompatibility complex (MHC) specific. Despite long survival, BALB/c hearts all showed evidence of parenchymal damage and vascular intimal thickening, a sign of chronic rejection. PMID- 10375077 TI - Regulatory function of human CD4+ cytolytic T lymphocytes. AB - Allograft rejection is mediated by both CD4+ and CD8+ T cells. The lytic function of the classic CD8+ cytolytic T lymphocytes (CTL) occurs through recognition of allogeneic major histocompatibility complex (MHC) class I on the surface of the graft. CD4+ CTL recognize MHC class II through a direct recognition pathway or an indirect pathway where MHC peptides are presented in the context of self MHC class II. Lytic CD4+ cells may destroy graft tissue or, we hypothesize, the indirect CD4+ T cell may down regulate CD8+ CTL by recognition of donor MHC peptides presented by self MHC class II expressed on CD8+ T cells. To define the role of CD4+ CTL in allograft outcome we used a CD4+ CTL that is MHC class II restricted, recognizing human leucocyte antigen (HLA)-A1 and HLA-B8 peptides in the context of HLA-DR4. This line (MDSxA1/B8) will lyse DR4+ B lymphoblastoid cells (LCL) pulsed with HLA-A1/B8 peptides (amino acids 60-84 of the alpha1 domain of the MHC class I molecule). These T cells will also lyse peptide-pulsed antigen-specific T cell clones, both CD4+ and CD8+, that express HLA-DR4. These clones must process and present the MHC class I peptides for recognition and lysis to occur. These results suggest a possible mechanism to explain allograft tolerance. Lytic CD4+ T cells, that recognize donor HLA peptides through an indirect antigen presentation pathway, down-regulate donor-specific CTL through peptide-specific lysis resulting in graft tolerance. PMID- 10375078 TI - Trafficking of APC from liver allografts of Flt3L-treated donors: augmentation of potent allostimulatory cells in recipient lymphoid tissue is associated with a switch from tolerance to rejection. AB - Livers transplanted across major histocompatibility complex (MHC) barriers in mice are normally accepted without recipient immune suppression, and induce a state of functional tolerance. However, markedly increasing functional dendritic cells (DC) in the 'passenger leucocyte' population by donor pretreatment with the hematopoietic growth factor Flt3-ligand (Flt3L; 10 microg/day for 10 days) results in acute allograft rejection. In this study, molecular, immunohistochemical and flow cytometric analysis of donor cell traffick into recipient lymphoid tissue 24 h after liver transplantation (C57BL/10 [H2b]-->C3H [H2k]) was performed. In addition, the capacity of donor-derived cells in these tissues to stimulate host T cell proliferation was examined. Reverse transcriptase polymerase chain reaction analysis revealed increases in donor genomic DNA in both thymi and spleens of mice given livers from Flt3L-treated donors compared to controls. Donor MHC class II+ (IAb+) cells in spleens were strikingly elevated (10-fold) in the former group. Two-colour flow cytometry revealed a similar increase in donor-derived H-2Kb+/I-Ab+ cells, and in the incidence of donor leucocytes expressing CD40, CD80, and CD86. CD11c+ DC comprised approximately 40% of the I-Ab+ cells in spleens of mice given livers from Flt3L-treated donors. These changes were associated with the presence, in spleens, of potent allostimulatory activity for naive recipient strain T cells, that was not observed in normal liver recipients. Elicitation of allograft rejection, associated with enhanced trafficking of stimulatory donor antigen presenting cells (APC), in particular DC, suggests that normal liver graft survival and tolerance induction may be linked to failure/counter-regulation of APC-driven stimulation of effective anti-donor T cell responses. PMID- 10375079 TI - Beneficial and detrimental human leucocyte antigen (HLA)-DR mismatches are not reflected by a differential effect of immunosuppressive drugs on the in vitro alloimmune response. AB - The introduction of molecular tissue typing techniques has led to an enormous increase in the number of human leucocyte antigen (HLA) alleles. This increasing polymorphism of the HLA antigens makes the selection of a well-matched unrelated donor a difficult task. Recent data suggest that some HLA mismatches are more immunogenic than others. This has led to the introduction of terms like beneficial or acceptable versus detrimental or taboo mismatches. The study considered whether the differential immunogenicity as reflected by graft survival studies can be detected in vitro as well. Mixed lymphocyte reaction (MLR) and primed lymphocyte tests (PLT) were performed with different HLA-DR mismatched combinations in the presence and absence of cyclosporine A and prednisolone. Differential effects of these immunosuppressive drugs were observed. Some reactions could easily be blocked by cyclosporine alone, whereas others need the addition of high doses of prednisolone as well before a significant inhibition was found. These differences were not only found between individual responders but also within one individual dependent on the stimulatory HLA-antigen involved. When the group of beneficial mismatches was compared with the group of detrimental mismatches, no differences were observed. Our data show that immunosuppressive drugs have a differential effect on in vitro alloimmune responses but these do not differentiate between beneficial and detrimental mismatches as defined by kidney graft survival. PMID- 10375080 TI - Differential Th1 and Th2 cell regulation of murine cardiac allograft acceptance by blocking cell adhesion of ICAM-1/LFA-1 and VCAM-1/VLA-4. AB - Administration of anti-intercellular adhesion molecule (ICAM)-1 monoclonal antibody (mAb) plus anti-lymphocyte function associated antigen (LFA)-1 mAb induces tolerance in murine cardiac transplantation, while anti-vascular cell adhesion molecule (VCAM)-1 mAb plus anti-very late antigen (VLA)-4 mAb administration prolongs graft survival, but leads to tolerance only in some cases. BALB/c mice hearts were transplanted into C3H/He recipients. Each combination of anti-VCAM-1 plus anti-VLA-4 mAbs (100 microg each/day, i.p.) or anti-ICAM-1 plus anti-LFA-1 mAbs (50 microg each/day, i.p.) was administered for 5 days. For control study, third group mice received daily with FK506 administration (1 mg/kg/day). The cardiac allografts and recipients' spleens were harvested on day 7; the expression of cytokines were detected using immunohistochemistry, reverse transcription-polymerase chain reaction (RT-PCR) and in situ RT-PCR. Th2 cytokines (IL-4 and IL-10) were markedly enhanced and Th1 cytokines (IFN-gamma and IL-2) were suppressed in recipients treated with anti ICAM-1 mAb plus anti-LFA-1 mAb, while poor Th2 cytokine expression allowed persistent Th1 cytokine expression in recipient mice with anti-VCAM-1 mAb plus anti-VLA-4 mAb treatment. Both Th1 and Th2 cytokine expression was suppressed in FK506-treated mice. It is concluded that immunological tolerance and prolonged graft survival induced by blocking cell adhesion is regulated by different cytokine expression. PMID- 10375081 TI - Introduction: extranodal lymphomas. PMID- 10375082 TI - Primary large cell lymphoma of the mediastinum. AB - Primary mediastinal (thymic) large B-cell lymphoma is a discrete clinicopathologic subtype of diffuse large cell lymphoma recognized in the revised European-American lymphoma classification. It is an uncommon but not rare tumor with worldwide distribution. For the clinician, the occurrence of this aggressive, invasive, yet localized neoplasm arising in an unusual site in a cohort of young adult patients (frequently women) in whom large cell lymphoma is infrequent is noteworthy. The pathologist is impressed both by the characteristic sclerosis and the unusual surface immunoglobulin (SIg)-negative B-cell phenotype of a tumor arising in a T-cell organ (thymus). The phenotype is that of a B-cell subset resident in the thymic medulla. The pattern of spread resembling that of extranodal lymphoma and the excellent response to appropriate combination chemotherapy and irradiation further support the discrete character of mediastinal large cell lymphoma, and provide a practical reason for recognizing the entity. Poorly chosen or executed primary therapy can lead to rapid tumor growth or regrowth with treatment failure and death, but a reassuringly high cure rate follows appropriate management. PMID- 10375083 TI - Testicular lymphoma. AB - Non-Hodgkin's lymphoma of the testis is an uncommon disease. It accounts for approximately 9% of testicular neoplasms. Despite this low overall incidence, however, it is the most common testicular malignancy in the elderly. Testicular lymphoma has a rather high incidence of bilateral involvement and a propensity for extranodal spread to the skin, subcutaneous tissue, CNS, lung, and Waldeyer's ring. Although intermediate-grade diffuse large B-cell lymphoma is the most common histologic pattern among primary testicular lymphoma, secondary infiltration of the testis, especially in high-grade Burkitt's lymphoma, is more prevalent. Although excellent results with a doxorubicin-containing chemotherapy regimen have been achieved in early-stage disease, patients with advanced disease have a grave prognosis. PMID- 10375084 TI - Primary non-Hodgkin's lymphoma of bone. AB - Primary non-Hodgkin's lymphoma of bone (PLB) constitutes approximately 5% of all extranodal non-Hodgkin's lymphoma (NHL) and 7% of primary bone tumors. The peak incidence for PLB is in the fifth decade, with a slight preponderance of males over females. The presenting symptoms usually consist of localized bone pain and occasionally a palpable mass. Most patients with PLB have B-cell tumors with a diffuse mixed-cell or diffuse large cell histology. While most patients present with early-stage disease, it is not clear whether such patients benefit from combined-modality therapy (CMT) consisting of radiation therapy (RT) and chemotherapy (CT) compared with either RT or CT alone. However, there is strong evidence that CMT is beneficial in the treatment of localized NHL, and these results might be applicable to the therapy for PLB. Nevertheless, only a phase III randomized, controlled clinical trial will determine whether CMT is superior to either CT or RT alone. PMID- 10375085 TI - Mycosis fungoides and the Sezary syndrome. AB - Mycosis fungoides (MF) and the Sezary syndrome are a group of extranodal non Hodgkin's lymphomas of T-cell origin with primary cutaneous involvement. The group distinguishes itself from other primary cutaneous T-cell lymphomas (CTCLs) by its unique clinical features and histopathology. In its early stages, it often resembles common benign dermatoses, and therefore, a definitive diagnosis can be delayed. The affected T cells are characterized by a predominant CD4+ phenotype with frequent loss of CD7 (pan-T-cell antigen) and often demonstrate T-cell receptor (TCR) rearrangement. The prognosis of patients with MF is highly dependent on the extent and type of skin involvement. The initial cutaneous presentation of MF can be patches, plaques, tumors, or erythroderma. Patients who present with limited patch/plaque disease have an outstanding prognosis with an overall long-term survival that is similar to the expected survival of a matched control population. It is exceedingly rare for patients who present with limited or generalized patch/plaque disease without peripheral lymphadenopathy to have extracutaneous involvement. Therefore, the staging evaluation differs for patients with MF versus patients with other non-Hodgkin's lymphomas and should be tailored to the clinical presentation. Patients who have tumorous or erythrodermic skin involvement have a less favorable prognosis, and patients who present with extracutaneous disease have a poor prognosis. There are multiple therapeutic options for patients with MF and the Sezary syndrome. Selection of a specific treatment plan is based primarily on the clinical stage of the disease. The primary therapy for patients with patch/plaque disease without extracutaneous involvement is a topical regimen, whereas chemotherapy or other aggressive systemic regimens are reserved for those with recalcitrant disease or extracutaneous involvement. There is no evidence that early aggressive systemic therapy is preferable to conservative therapy in the management of limited disease. There are newer combination topical and/or systemic regimens that result in an improved clinical response and possibly a prolonged response duration. For advanced disease, standard therapies are often palliative and successful clinical response is often very short-lived. Therefore, all patients with recalcitrant or extracutaneous disease should be considered for newer investigative therapies. PMID- 10375086 TI - Primary cutaneous lymphomas other than mycosis fungoides. AB - Primary cutaneous lymphomas present in and are confined to the skin with no evidence of extracutaneous disease. The skin is the second most common extranodal site involved by primary lymphoma; 50% are mycosis fungoides (MF)-type cutaneous T-cell lymphoma, with the remainder being peripheral T-cell lymphoma (25%) and B cell lymphoma (25%). The diagnosis of non-MF primary cutaneous lymphomas differs from that of nodal lymphomas: (1) presentation in the skin more often predicts outcome than histology, (2) immunophenotyping and immunogenotyping studies show differences in chromosomal translocations, cell-surface antigen expression (T cell receptor [TCR] and immunoglobulin [Ig] heavy and light chains), and oncogene expression, (3) involvement of structural compartments of the skin (epidermis, periadnexal or adventitial dermis, interstitial dermis, and subcutis) aids differential diagnosis in place of nodal architecture, and (4) cytokine and extracellular matrix environments may influence behavior of cutaneous lymphomas. Diagnosis often requires coordinated evaluation of clinical history, immunohistochemistry on paraffin and frozen sections of skin biopsies, and molecular analysis. Classification of primary cutaneous lymphomas by a combined histologic type and clinical behavior is useful. PMID- 10375087 TI - Primary pulmonary lymphoma. AB - Lymphoproliferative diseases affecting the lung occur over a broad clinical and pathologic spectrum. The clinical presentations and radiologic findings are nonspecific, entailing broad differential diagnoses. Accurate diagnosis requires adequate tissue sampling with appropriate ancillary pathologic studies. The recent delineation of new pathologic entities such as low-grade malignant lymphoma of mucosa-associated lymphoid tissue (MALT type) has aided in the understanding of the pathophysiology, clinical course, and management of patients with pulmonary lymphoma. Significant observations have been made in the clinical management and treatment of these disorders. PMID- 10375088 TI - Primary thyroid lymphoma. AB - The clinical and pathologic spectrum of lymphoproliferative disorders affecting the thyroid is diverse and must be differentiated from benign thyroiditis and carcinoma. The clinical presentations include an enlarging neck mass, but patients may also present with symptoms of dysphagia, hoarseness and choking, or a cold thyroid nodule. The histopathologic interpretation requires adequate tissue sampling and proper pathologic interpretation. The recent delineation of new pathological entities such as low-grade malignant lymphoma of mucosa associated lymphoid tissue (MALT) type has aided in the understanding of the clinical course and management of patients with lymphoma. Advances have been made in the clinical management and treatment of these disorders. Surgical resection of the thyroid mass is not routinely part of the management strategy. The management of low-grade lymphoproliferative disorders of MALT type may include radiation therapy, oral chlorambucil, or intravenous chemotherapy (cyclophosphamide, vincristine, and prednisone). The management of diffuse large B-cell lymphoma is combined-modality therapy with radiation and cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) chemotherapy. PMID- 10375089 TI - Lymphoma of the gastrointestinal tract. AB - Non-Hodgkin's lymphoma (NHL) of the gastrointestinal (GI) tract accounts for 4% to 20% of all NHLs and is the most common extranodal site of presentation. The stomach is the major organ involved by GI lymphoma. Helicobacter pylori infection, immunosuppression after solid-organ transplantation, celiac disease, inflammatory bowel disease, and human immunodeficiency virus (HIV) infection may be risk factors for GI lymphoma. A significant proportion of gastric lymphomas are of low-grade histology and arise from mucosal-associated lymphoid tissue (MALT). Such MALT lymphomas may be associated with H. pylori infection and may undergo complete regression following eradication of H. pylori. Lymphoma of the small bowel, colon, and rectum may also occur, but are less common than gastric lymphoma. Distinct clinicopathologic entities, such as primary intestinal T-cell lymphoma, immunoproliferative small intestinal disease, and multiple lymphomatous polyposis have been described. Surgery, radiation therapy, and chemotherapy have been used in the treatment of GI lymphomas. However, the optimal management of these lymphomas has never been determined by prospective randomized clinical trials. Such trials by cooperative groups are needed to answer many of the vital unanswered questions concerning extranodal lymphomas of the GI tract. PMID- 10375090 TI - Lymphomas of the head and neck. AB - Lymphomas of the head and neck arise in Waldeyer's ring, the salivary glands, nasal cavity, paranasal sinuses, thyroid gland, and orbit. Though anatomically in close proximity, lymphomas arising in these sites have distinct clinical characteristics. Factors that appear to influence the pattern of disease include concurrent conditions, such as Sjogren's syndrome, and geographic factors, particularly with regard to nasal lymphomas. The treatment and prognosis of patients with head and neck lymphoma depends on the histologic grade of disease and extent of involvement at time of presentation. Most lymphomas are of intermediate-grade histology and early stage at presentation. A thorough understanding of clinical disease patterns and treatment options will allow the optimum management of these patients. PMID- 10375091 TI - Primary CNS lymphoma. AB - Primary CNS lymphoma (PCNSL) is seen with increasing frequency in both immunocompetent and immunocompromised patients. Radiation therapy has historically been standard treatment for this disease, resulting in complete remissions in the majority of patients, but with most patients relapsing and dying of the disease in less than 2 years. The use of chemotherapy appears to be changing the natural history of this tumor, with significantly prolonged survival in some groups of patients. The optimal agents, doses, schedules, routes of administration, and need for radiation therapy, however, remain undefined. In this review, some of the questions regarding the management of PCNSL are addressed and recommendations are made regarding the design of future regimens. PMID- 10375092 TI - Lymphomas of the breast: pathology and clinical behavior. AB - Lymphocytes are present in normal breast. A lymphocytic mastopathy characterized by a lymphocytic infiltrate within the breast epithelium has been described, but its relevance as a precursor lesion of mucosa-associated lymphoid tissue (MALT) type lymphoma of the breast is uncertain. Lymphomas of the breast are uncommon, and a broad variety of histologic types have been reported. The majority are B cell lymphomas, and the most common type is diffuse large B-cell lymphoma (40% to 70%). MALT-type lymphoma is a distinct subgroup of primary lymphoma of the breast with a reported incidence between 0% and 44% and is characterized by indolent behavior and good prognosis. Burkitt's or Burkitt-like lymphoma can bilaterally involve the breast of a young pregnant or lactating woman and typically behaves aggressively. Primary breast lymphomas behave similarly to lymphomas of similar histologic types and stages presenting at other sites. Treatment of primary breast lymphomas does not include surgery, but is typically based on local radiotherapy, often combined with systemic chemotherapy. PMID- 10375093 TI - Soft tissue sarcoma: opening the door to advances in diagnosis and treatment. PMID- 10375094 TI - The evaluation and treatment of patients receiving radiation therapy for carcinoma of the esophagus: results of the 1992-1994 Patterns of Care Study. AB - BACKGROUND: For the first time, a Patterns of Care Study (PCS) was conducted in 1992-1994 to determine the national practice standards in evaluating and treating patients with esophageal carcinoma and to determine the degree to which clinical trials have been incorporated into national practice. METHODS: A national survey of 61 institutions using 2-stage cluster sampling was conducted, and specific information was collected on 400 patients with squamous cell carcinoma or adenocarcinoma of the thoracic esophagus who received radiation therapy (RT) as part of definitive or adjuvant management of their disease. Patients were staged according to a modified 1983 American Joint Committee on Cancer staging system. Chi-square tests for significant differences between academic and nonacademic institutions for a particular variable were performed. RESULTS: The median age of patients was 66.7 years (range, 26-89 years); 76.5% were male and 23.5% were female. Karnofsky performance status was > or = 80 for 88.3% of patients. Squamous cell carcinoma was diagnosed in 61.5% and adenocarcinoma in 36.8%. Fifteen percent were Clinical Stage (CS) I, 39.5% CS II, and 29.5% CS III. Evaluative procedures included endoscopy (>93%), computed tomography (CT) of the chest (86%), CT of the abdomen (75%), esophagography (68.5%), and endoscopic ultrasound (3.5%). Endoscopic ultrasound and CT of the chest were performed significantly more frequently at academic than nonacademic facilities (6.1% vs. 1.0% and 91.9% vs. 81.3%, respectively). Three-quarters of all patients received chemotherapy and RT and 62.5% received concurrent chemotherapy and RT as part of their treatment. Treatments included chemotherapy plus RT (54.0%), RT alone (20.3%), preoperative chemotherapy + RT (13.3%), postoperative chemotherapy + RT (7.7%), postoperative RT (3.5%), and preoperative RT (1.2%). The chemotherapeutic agents most frequently used were 5-fluorouracil (84%), cisplatin (64%), and mitomycin (9%); academic instututions used cisplatin significantly more often and mitomycin significantly less often than nonacademic institutions. Brachytherapy was used in 8.5% of cases. The median total dose of external beam radiation was 50.4 gray and the median dose per fraction was 1.8 gray. CONCLUSIONS: This study establishes the national benchmarks for the evaluation and treatment of patients with esophageal carcinoma at radiation facilities in the U.S. It also indicates that the majority of patients given RT as a component of treatment for esophageal carcinoma receive chemoradiation rather than RT alone, as supported by clinical trials. Although some differences in the evaluation of esophageal carcinoma were noted between academic and nonacademic facilities, there was no difference in the frequency of use of chemoradiation versus RT by facility type. PMID- 10375095 TI - Patients younger than 40 years with gastric carcinoma: Helicobacter pylori genotype and associated gastritis phenotype. AB - BACKGROUND: In the general population, Helicobacter pylori (H. pylori), particularly the cagA positive strain, has been associated with intestinal-type gastric carcinoma. Gastric carcinomas are rarely observed in patients age < or = 40 years. Host-related factors have been thought to be more important than environmental agents in these early-onset cancers. The aim of this study was to ascertain the possible role of H. pylori infection and that of cagA positive strains in the development of gastric carcinoma in these young patients. METHODS: In this case-control study, 105 gastric carcinoma patients (male-to-female ratio = 1.1; mean age, 34.4 years; range, 16-40 years) and an equal number of controls (matched for gender and age) were retrospectively selected from the same geographic area. The phenotypes of gastritis and H. pylori were histologically assessed, and the presence of the ureC gene, which is indicative of H. pylori infection, and the cagA genotype were determined by polymerase chain reaction. Gastric carcinoma risk was calculated by both univariate and multivariate statistical methods, taking into account the cancer phenotype, the gastritis phenotype detected in both patients and controls, and the H. pylori genotype. RESULTS: For 74 diffuse and 31 intestinal gastric carcinomas, multivariate logistic regression analysis produced results consistent with those of univariate statistical tests, showing a significant association between gastric carcinoma and both H. pylori infection (odds ratio [OR] = 2.79; 95% confidence interval [CI] = 1.52-5.11) and cagA positive status (OR = 2.94; 95% CI = 1.56-5.52). CONCLUSIONS: In young Italian patients with gastric carcinoma, the significant association with cagA positive H. pylori infection suggests that the bacterium has an etiologic role in both diffuse-type and intestinal-type gastric carcinoma. PMID- 10375096 TI - Influence of selection criteria on mutation detection in patients with hereditary nonpolyposis colorectal cancer. AB - BACKGROUND: Hereditary nonpolyposis colorectal cancer (HNPCC) is linked genetically to mutations in DNA mismatch repair (MMR) genes. Because a deficiency in MMR does not predict a specific phenotype, the original selection criteria may be too restrictive in identifying additional families. The current study was performed to determine whether a relaxation of the Amsterdam criteria (AC) could be applied to identify more families associated with DNA MMR. METHODS: Twenty eight unrelated Swiss families (15 complying with the AC and 13 fulfilling extended criteria [EC] to include other tumors of the HNPCC spectrum as well) were screened for mutations in the MMR genes hMSH2 and hMLH1, using single stranded conformation polymorphism and direct DNA sequencing. Microsatellite instability (MSI) was determined in 14 families. A comparison was made between the phenotypic characteristics of the mutation positive and mutation negative families. RESULTS: Ten AC families (67%) harbored germline mutations in hMLH1 (6 kindreds) or hMSH2 (4 kindreds). In none of the EC kindreds could an unambiguous disease-causing mutation be identified. Seven of eight AC families were found to display MSI whereas all colorectal carcinomas (CRC) in eight EC kindreds were MSI stable. CRC patients from mutation positive families had an earlier age at diagnosis (44 years vs. 49 years) and appeared to have a better survival (11.1 years vs. 7.7 years). CONCLUSIONS: Extending the AC to include extracolonic tumors of the HNPCC spectrum results in a very low mutation detection rate for hMSH2 and hMLH1. The EC families appear to represent an alternative genetic entity not necessarily related to DNA MMR gene mutations because they do not display MSI. PMID- 10375097 TI - Colorectal carcinoma among ethnic Chinese in Singapore: trends in incidence rate by anatomic subsite from 1968 to 1992. AB - BACKGROUND: Recent epidemiologic studies have suggested that the anatomic distribution of colorectal carcinoma may have undergone a distal to proximal shift over several decades, which has been attributed variously to environmental and genetic factors as well as preventive intervention. METHODS: Trends in subsite distribution and the incidence rate of colorectal carcinoma among Chinese in Singapore between 1968 and 1992 were explored using data from the Singapore Cancer Registry (n = 10,489). Age-standardized incidence rates were computed and compared further using age-period-cohort models by subsite and gender. RESULTS: The proportion of lesions in the distal colon was found to have increased from 23.2% to 24.4% whereas that for the proximal colon and rectum were fairly consistent over the past 25 years. Our results also showed that age-standardized rates have doubled in proximal lesions (2-3% annually) and more than doubled in distal lesions (3-4% annually) whereas rates in rectal carcinoma have shown a slight increase or stability over time. The patterns of change in all subsite tumors could be attributed to a significant birth cohort effect. CONCLUSIONS: The results of the current study suggest that incidence rates have increased rapidly with no distal to proximal shift observed among ethnic Chinese in Singapore over the past 25 years. The pattern of change differs from findings reported in high incidence countries such as the U. S. and parts of Europe, suggesting that the preventive intervention and early diagnostic capabilities that may have played an important role in these countries have had less effect in Asia. The rapid overall increase in the incidence rate of colon carcinoma supports the role of dietary and other environmental factors as possible risk factors. PMID- 10375098 TI - Serum interleukin-6 level reflects the tumor proliferative activity in patients with colorectal carcinoma. AB - BACKGROUND: Interleukin (IL)-6 plays a central role as a differentiation and growth factor of tumor cells. It may be hypothesized that increased serum IL-6 derives from the tumor tissue, and that serum IL-6 levels consequently reflect the biologic characteristics of the tumor. The authors investigated the association between the serum levels of IL-6 in colorectal carcinoma patients and the biologic characteristics of the tumor as well as clinicopathologic status of the patients. METHODS: Serum and tissue levels of IL-6 in 70 patients were determined using an immunoradiometric assay. Expression of IL-6 and IL-6 receptor were evaluated immunohistochemically. The proliferative activity of the tumor was assessed by nuclear Ki-67 labeling index. RESULTS: The concentration of serum IL 6 in the patients was significantly higher than that in normal controls. The tissue concentration of IL-6 in tumor tissue was significantly higher than that in normal mucosa, and was correlated strongly with the serum IL-6 concentration. The serum IL-6 concentration was correlated with clinicopathologic parameters, including liver metastases and tumor size, and with proliferative activity of the tumor as assessed by the Ki-67 labeling index. In the patients with Stage I or II tumors, the preoperative serum IL-6 concentration was reduced significantly 3 months after surgery. Immunohistochemically, tumor tissues that expressed IL-6 immunoreactivity had a higher incidence of expression of IL-6 receptor immunoreactivity. CONCLUSIONS. Serum IL-6 concentration reflects IL-6 concentration in the tumor component and may reflect the proliferative activity of the tumor in patients with colorectal carcinoma. PMID- 10375099 TI - A multicenter evaluation of intensified, ambulatory, chronomodulated chemotherapy with oxaliplatin, 5-fluorouracil, and leucovorin as initial treatment of patients with metastatic colorectal carcinoma. International Organization for Cancer Chronotherapy. AB - BACKGROUND: The combination of 5-fluorouracil (5-FU), leucovorin (LV), and oxaliplatin (I-OHP) was shown to be both more active against metastatic colorectal carcinoma and better tolerated if the drug delivery rate was chronomodulated according to circadian rhythms rather than constant. This allowed the authors to intensify the three-drug chronotherapy regimen and to assess its activity as the initial treatment of metastatic colorectal carcinoma patients in ten centers from four countries. METHODS: Patients with previously untreated and inoperable measurable metastases from colorectal carcinoma received a daily administration of chronomodulated 5-FU (700 mg/m2/day, peak delivery rate at 04:00 hours), LV (300 mg/m2/day, peak delivery rate at 04:00 hours), and 1-OHP (25 mg/m2/day, peak delivery rate at 16:00 hours) for 4 days every 14 days. Intrapatient escalation of 5-FU dose was performed if toxicity was less than World Health Organization (WHO) Grade 2. RESULTS: Of 90 enrolled patients, 35 had a WHO performance status of 1 or 2; 49 had metastases in > or = 2 organs. The liver was involved in 79 patients, 30 of whom had clinical hepatomegaly. The main dose-limiting toxicities were WHO modified Grade 3 or 4 diarrhea (41% of patients, 8.2% of courses), stomatitis (30% of patients, 5.1% of courses), and Grade 2 cumulative peripheral sensory neuropathy (19% of patients after 12 courses). Two patients died with severe gastrointestinal toxicity. Using the intent-to-treat method, the overall objective response rate was 66% (95% confidence limits, 56-76%). Surgical removal of previously inoperable metastases was successful in 31 patients (34%). Histologic necrosis of metastases was >90% in 7 patients and complete in 1 patient. The median progression free survival and survival durations were 8.4 months (range, 5.9-10.9 months) and 18.5 months (range, 13.2-23.8 months), respectively, with 38% of the patients alive at 2 years of follow-up. CONCLUSIONS: The objective response rate appeared to be approximately 3-fold as high as that achieved with current 5-FU-based regimens and translated into an approximately 50% increase in median survival. The hypothesis that this intensified, ambulatory, chronotherapy regimen can increase survival currently is being investigated in a multicenter randomized study conducted by the European Organization for Research and Treatment of Cancer Chronotherapy Study Group. PMID- 10375100 TI - Influence of p53 status on prognosis in preoperatively irradiated rectal carcinoma. AB - BACKGROUND: Even when they are analogous in microscopic and macroscopic appearance, tumors vary in their response rates to radiotherapy. Cell culture and xenograft experiments with colorectal cell lines have demonstrated that wild-type p53 increases radiosensitivity. Hence, the authors investigated, in a well defined population of patients treated at the same institution, whether p53 status was a prognostic factor in preoperatively irradiated rectal carcinoma patients. METHODS: The p53 status of rectal adenocarcinomas was examined immunohistochemically (with monoclonal antibody DO-1) in preirradiated biopsy samples (n = 100) and corresponding postirradiated resected specimens (n = 97). The mean follow-up was 73.2 months (median, 71.3 months; range, 4.3-157 months). Statistical analysis was performed using the SPSS program (SPSS, Chicago, IL). RESULTS: p53 protein expression was detected in 55 of 100 biopsy samples (> or = 5% nuclear staining). There was essentially no difference in p53 expression between biopsy samples and corresponding resected specimens (54 of 97 vs. 55 of 97). In univariate analysis, p53 immunoreactivity of biopsy samples did not correlate with age, gender, tumor location, TNM stage, pT category, pN category, or histologic grade. Unlike clinicopathologic variables, p53 expression did not have a statistically significant association with local recurrence free, disease free, or overall survival in either univariate (P = 0.91, 0.18, and 0.17, respectively) or multivariate analysis. CONCLUSIONS: In contrast to cell line studies, this immunohistochemical study demonstrates that p53 status is not useful as a prognostic marker in preoperatively irradiated rectal carcinoma. PMID- 10375101 TI - Local recurrence after vertical partial laryngectomy, a conservative modality of treatment for patients with Stage I-II squamous cell carcinoma of the glottis. AB - BACKGROUND: Based on an inception cohort of 103 patients who had local recurrence (Group I) and a witness group of 311 patients who achieved local control (Group II) after vertical partial laryngectomy for Stage I-II glottic carcinoma, the current retrospective study documented the consequences and management of local recurrence. METHODS: Three hundred two patients (97.1%) in Group II and all 103 patients (100%) in Group I were followed until death or for a minimum of 10 years. Statistical analysis of survival, lymph node control, and distant metastasis was based on the Kaplan-Meier product limit method. RESULTS: The 10 year actuarial survival estimate was 30.8% for Group I patients and 63.1% for Group II patients. Survival was statistically more likely to be reduced in Group I patients (P < 0.0001) than in Group II patients. The percentage of patients who died of their initial disease was 44.6% in Group I and 6.3% in Group II. The 10 year actuarial lymph node control estimate was 70.2% for Group I and 96.1% for Group II. Lymph node recurrence was statistically more likely to occur in Group I patients than in Group II patients (P < 0.0001). The 10-year actuarial estimate for patients without distant metastasis was 80.2% for Group I and 96.7% for Group II. Distant metastasis was statistically more likely to occur in Group I patients than in Group II patients (P < 0.0001). Salvage treatment was unsuitable for 4.7% of patients with local recurrence; for other patients, it yielded a 86.7% local control rate, a 21.4% laryngeal preservation rate, a 4.5% death rate, and an 11.2% rate of incidence of severe complications. CONCLUSIONS: Among patients with Stage I-II glottic carcinoma managed with vertical partial laryngectomy, local recurrence results in a reduced rate of survival as well as a high rate of necessity for salvage total laryngectomy. PMID- 10375102 TI - Multiple chondromatous hamartomas of the lung: a case report and review of the literature with special reference to Carney syndrome. AB - BACKGROUND: Multiple chondromatous hamartomas of the lung, which are very rare, are a feature of Carney syndrome. The relation between the two entities is not clear. METHODS: A patient with multiple chondromatous hamartomas of the lung is described in this article. The literature was reviewed with special reference to the relation between multiple chondromatous hamartomas of the lung and Carney syndrome as well as the triad of gastric epithelioid leiomyosarcoma, functioning extra-adrenal paraganglioma, and pulmonary chondroma. RESULTS: A total of 15 cases of multiple chondromatous hamartomas of the lung have been published worldwide. Two cases exhibited two other features of Carney syndrome, namely, gastric leiomyogenic neoplasms and extra-adrenal paragangliomas, and three other cases demonstrated only gastric leiomyomatous neoplasms. These five patients were all young females. CONCLUSIONS: Some patients with multiple chondromatous hamartomas of the lung have a history of Carney syndrome. Patients with multiple chondromatous hamartomas require further examination of other sites, particularly the stomach and nervous system. PMID- 10375103 TI - A phase I/II trial of neoadjuvant chemotherapy with cisplatin and vinorelbine followed by accelerated irradiation for patients with inoperable nonsmall cell lung carcinoma. AB - BACKGROUND: Both locoregional and distant disease control remains poor in the treatment of Stage III nonsmall cell lung carcinoma (NSCLC). This trial was conducted to evaluate the tolerance and responses of patients with NSCLC given a neoadjuvant regimen of cisplatin and vinorelbine chemotherapy followed by accelerated thoracic radiotherapy. METHODS: Forty-two patients with Stage IIIA and IIIB NSCLC were entered into the study. Treatment consisted of cisplatin 100 mg/m2 given on Days 1 and 29 and vinorelbine 30 mg/m2 given weekly for 5 weeks, with a planned 50% dose reduction to 15 mg/m2 planned for Week 2. This was followed by thoracic irradiation of 60 gray (Gy) in 30 fractions of 2 Gy over 4 weeks (once daily during Weeks 1 and 2 and twice daily during Weeks 3 and 4). RESULTS: With a median follow-up time of 12.2 months (27-65 months for survivors), the median survival was 12.2 months (16.6 months for patients with no prior weight loss and 7.8 months for those with prior weight loss). The response rate after induction chemotherapy was 46.1%, increasing to 74.4% after radiation therapy (8 complete responses and 21 partial responses). The rate of progression was 13 of 18 (72%) for those who responded to chemotherapy (4 distant, 9 local) and 18 of 21 (86%) for those who did not respond to chemotherapy (14 distant, 7 local). The most frequent acute Grade 3 toxicity was nausea (21.4%). CONCLUSIONS: Accelerated thoracic irradiation after induction chemotherapy is well tolerated and yields therapeutic results that compare favorably with those reported for other regimens of chemotherapy and standard fractionated radiotherapy. The data from this study suggest that the responses of patients with clinically apparent disease to induction chemotherapy might indicate a likelihood of controlling microscopic distant metastases. PMID- 10375104 TI - Thrombospondin-1 expression and clinical implications in malignant pleural mesothelioma. AB - BACKGROUND: The role of thrombospondin-1 (TSP-1) in tumor angiogenesis and progression is controversial. The authors assessed the impact of TSP-1 as a prognostic indicator in malignant pleural mesothelioma (MPM). METHODS: TSP-1 expression was assessed by reverse transcriptase-polymerase chain reaction using 5 normal pleural samples, 78 MPM tumors, and 43 surrounding normal lung samples. In MPM tumors, vascular endothelial growth factor (VEGF) expression also was examined. Differences between different valuables were analyzed using the Mann Whitney U test. Survival curves were obtained by the Kaplan-Meier method and the survival rate was assessed by the log rank test. RESULTS: TSP-1 was highly expressed in 74 of the 78 MPM tumors (95%) with a mean value of 2.27 +/- 0.42 compared with normal pleura (0.50 +/- 0.06) and surrounding normal lung (0.96 +/- 0.20) (P = 0.05 vs. normal pleura and P = 0.0006 vs. surrounding normal lung). The mean TSP-1 expression was significantly greater in high VEGF-expressing tumors (2.63 +/- 0.51) compared with low VEGF-expressing tumors (1.17 +/- 0.39; P < 0.0001) and TSP-1 expression was lower in patients with TNM Stage III/IV disease (n = 60) (1.85 +/- 0.37) than in patients with Stage I/II disease (n = 13) (4.46 +/- 1.74) (P = 0.025). The TSP-1 expression levels in tumors with lymph node metastases were significantly lower than in those without lymph node metastases (P = 0.0305). Although high TSP-1 expression was associated with good prognosis in patients with low VEGF-expressing tumors, TSP-1 itself appeared to have no overall impact on survival. The methylation status of a CpG island associated with the TSP-1 promoter was evident in MPM tumor samples despite high levels of TSP-1 mRNA expression. CONCLUSIONS: TSP-1 is overexpressed in MPM tumors but its expression is of little value as a prognostic indicator in MPM. However, the relations between TSP-1 and VEGF in MPM merit further investigation for possible innovative therapeutic interventions. PMID- 10375105 TI - A Phase II study of gemcitabine in patients with malignant pleural mesothelioma. European Organization for Research and Treatment of Cancer Lung Cancer Cooperative Group. AB - BACKGROUND: Gemcitabine has shown activity in patients with less chemosensitive solid tumors. Phase II screening of novel drugs is an accepted method with which to investigate new therapies in malignant mesothelioma. The European Organization for Research and Treatment of Cancer-Lung Cancer Cooperative Group has performed several sequential Phase II trials of new agents for the treatment of mesothelioma over the last 10 years. METHODS: Twenty-seven chemotherapy-naive patients with histologically proven malignant mesothelioma were treated with gemcitabine as a 30-minute intravenous administration of 1250 mg/m2 on Days 1, 8, and 15 of a 28-day cycle. Therapy continued for up to ten cycles unless disease progression or excessive toxicity mandated discontinuation. RESULTS: With a median relative dose intensity of 96%, toxicity was mild and neutropenia of > or = Grade 3 (according to National Cancer Institute criteria) occurred in 30% of patients, without episodes of febrile neutropenia. One case of hemolytic-uremic syndrome, most likely related to gemcitabine use, was observed. Overall, 2 objective responses were observed (response rate of 7%; 95% confidence interval, 1-24%). The median survival was 8 months. CONCLUSIONS: At the prescribed dosage and schedule, single agent gemcitabine appears to have limited activity in chemotherapy-naive patients with malignant pleural mesothelioma. PMID- 10375106 TI - Thrombotic microangiopathy associated with interferon therapy for patients with chronic myelogenous leukemia: coincidence or true side effect? AB - BACKGROUND: Interferon-alpha (rIFN-alpha) is an established therapy for patients with myeloproliferative disorders. Unusual immune-mediated side effects have been associated with rIFN-alpha therapy. The association of rIFN-alpha therapy with hemolytic uremic syndrome (HUS) and thrombotic thrombocytopenic purpura (TTP) has been reported infrequently. METHODS: Two patients with chronic myelogenous leukemia (CML) treated with rIFN-alpha-based regimens at the University of Texas M. D. Anderson Cancer Center developed thrombotic microangiopathy (HUS/TTP). The course of their disease is described. A third patient who developed renal failure while receiving rIFN-alpha therapy and had no other causative factor for his renal failure is also described. RESULTS: The patients were ages 24, 49, and 36 years, and they had received rIFN-alpha therapy for 37, 67, and 92 months, respectively, prior to the development of the disorder. One patient had discontinued rIFN-alpha 1 month before the event because of presumed rIFN-alpha related cardiomyopathy. Two patients received hydroxyurea and cytarabine as part of their therapy. No patient was receiving any medication known to be associated with HUS/TTP. None had a history of diarrheal illness, but Escherichia coli OH157.H7 was grown from the stool of one patient. Two patients responded to plasmapheresis with normalization of counts and other indices, but both developed renal failure and became dependent on dialysis. One patient had evidence of disease progression and died of multiorgan failure. The third patient required dialysis for 18 months but is currently off dialysis; this patient has some residual renal impairment. CONCLUSIONS: Although no definitive association between rIFN-alpha therapy and thrombotic microangiopathies can be concluded from these data, these and other previously reported cases suggest that HUS/TTP is a rare side effect of rIFN-alpha therapy that should be managed in the standard fashion. Hypotheses regarding the mechanism underlying this association are discussed in this article. PMID- 10375107 TI - Chemotherapy for patients with locally advanced or metastatic Merkel cell carcinoma. AB - BACKGROUND: Merkel cell carcinoma (MCC) is a highly malignant skin neoplasm. Regional lymph node and distant metastasis occur in 20-52% of patients. The role of chemotherapy in the treatment of patients with this rare tumor is unclear. METHODS: An exhaustive analysis of the literature (1980-1995) describing chemotherapy for patients with locally advanced or metastatic MCC was performed. All available published medical records (n = 101 patients) were entered in a database. In addition, data on six additional patients given chemotherapy during this time frame in Lyon, France, were included in the database. RESULTS: For the 107 patients, the overall objective response rate to first-line chemotherapy was 61% (61 of 101 evaluable patients). The response rate was 57 % (41 of 72) for patients with metastasis and 69% (20 of 29) for patients with locally advanced tumors. No clinical parameter was found to be correlated to response to chemotherapy. A high rate of toxic death during first-line treatment (n = 7.7%) was reported for these patients. The median overall survival from the date of chemotherapy initiation was 9 months for patients with metastasis and 24 months for patients with locally advanced tumors. The projected overall survival at 3 years was 17% for patients with metastasis and 35% for patients with locally advanced tumors. Progression after first-line chemotherapy was associated with significantly worse survival for patients with metastasis. Rates of response to second-line (n = 33) and third-line (n = 10) chemotherapy were 45% and 20%, respectively. CONCLUSIONS: MCC is chemosensitive but rarely chemocurable in patients with metastasis or locally advanced tumors. A high incidence of toxic death due to chemotherapy is reported in the literature. PMID- 10375108 TI - Synovial sarcoma: identification of low and high risk groups. AB - BACKGROUND: Synovial sarcoma, one of the most common soft tissue sarcomas that occur in adolescents and young adults, is generally viewed and treated as a high grade sarcoma. However, the authors' own experience and some previous studies have indicated that it has a wide spectrum of biologic behavior and that low and high risk subgroups of patients with synovial sarcoma can be identified. METHODS: A total of 121 consecutive patients with synovial sarcoma (including 66 males and 55 females ages 9-74 years), treated primarily or secondarily at 2 large referral centers for musculoskeletal tumors, were included in a statistical analysis conducted to identify independent prognostic factors. RESULTS: There were local recurrences in 38 patients (31%), usually after inadequate primary surgery outside the referral centers; 64 patients (54%) developed metastasis, primarily to the lungs. The estimated 5-, 10-, and 15-year survival rates were 60%, 50%, and 45%, respectively (the mean follow-up for surviving patients was 9.8 years, with a range of 1-30 years). In multivariate analysis, independent risk factors for local recurrence included larger tumor size and primary surgical resection outside the referral center. Independent risk factors for metastasis were older patient age, tumor with poor histologic differentiation, and tumor necrosis. For tumor-related death, the independent risk factors were older patient age, tumor with poor histologic differentiation, and larger tumor size. Local recurrence was associated with a 3.66-fold increased risk of tumor-related death. A low risk group (patient age <25 years, tumor size <5 cm, and no histologic evidence of poorly differentiated tumor) with 88% overall disease free survival was identified, as was a high risk group (patient age > or = 25 years, tumor size > or = 5 cm, and poorly differentiated tumor) with 18% overall disease free survival (P < 0.001). CONCLUSIONS: The identification of low and high risk synovial sarcoma patients indicates that synovial sarcomas are not uniformly high grade tumors. It also indicates that treatment strategies should be modified for these risk groups. Adequate primary surgery is essential to both local control and outcome for synovial sarcoma patients. PMID- 10375109 TI - Complete regression of MX-1 human breast carcinoma xenografts after targeted chemotherapy with a cytotoxic analog of luteinizing hormone-releasing hormone, AN 207. AB - BACKGROUND: Receptors for luteinizing hormone-releasing hormone (LH-RH) are found in about 50% of human breast carcinomas. A highly potent cytotoxic agent, 2 pyrrolinodoxorubicin (AN-201), was linked to the agonist [D-Lys6]LH-RH to form a cytotoxic LH-RH analog, AN-207, that can be targeted to LH-RH receptors on breast carcinomas. METHODS: Nude mice bearing MX-1 hormone-independent, doxorubicin resistant human breast carcinomas were injected intravenously with vehicle (control), 250 nmol/kg doses of AN-201, AN-207, or an unconjugated mixture of AN 201 and [D-Lys6]LH-RH. Tumor growth and changes in hematologic parameters were evaluated. Receptors for LH-RH were investigated by radioreceptor assays, and the expression of their mRNA was determined by reverse transcriptase-polymerase chain reaction. RESULTS: AN-207 caused complete regression of MX-1 tumors in all 10 animals, and they were still tumor free 60 days after treatment. In contrast, after therapy with AN-201 or the mixture of AN-201 and [D-Lys6]LH-RH, the regression of most MX-1 tumors was only transitory. AN-201 caused the death of 1 of the 10 animals and significantly greater leukopenia than AN-207, which produced no toxic deaths. Radioreceptor assays revealed high affinity binding sites for LH-RH on tumor cell membranes. The expression of mRNA for LH-RH receptors also was found in tumors. CONCLUSIONS: The results of this study indicate that powerful, targeted cytotoxic LH-RH analogs such as AN-207 could be considered for the treatment of human breast carcinomas that possesses receptors for LH-RH. PMID- 10375110 TI - Induction of apoptosis and suppression of clonogenicity of ovarian carcinoma cells with estrogen mustard. AB - BACKGROUND: This study was conducted to evaluate whether estramustine (estrogen mustard [EM]) is a promising alternative in the treatment of patients with epithelial ovarian carcinoma (OVCA). EM is a microtubule-associated protein (MAP) specific antimicrotubule agent with low toxicity. METHODS: The authors investigated the ability of EM to induce apoptosis and suppress colony formation of OVCA cells. Paclitaxel was used as a positive control. DNA electrophoresis and terminal deoxynucleotidyl dUTP-X nick end labeling (TUNEL) assays were used to detect internucleosomal DNA fragmentation. Flow cytometry was used to identify apoptotic cells and disturbance of the cell cycle of EM-treated OVCA cells further. Quantitation of detached cultured cells also provided a relative estimate of the apoptotic response of OVCA cells to treatment with EM. The colony formation assay was used to evaluate the effects of EM on clonogenicity. RESULTS: The effects of EM on four OVCA cell lines in culture were highly similar to those of paclitaxel in causing apoptosis and inhibiting clonogenicity. DNA electrophoresis and TUNEL assays showed that EM induced internucleosomal DNA fragmentation in OVCA cells. Flow cytometry showed changes typical of apoptotic changes and cell cycle block and synchronization at the G2/M-phase. Counting of detached cells showed a log-dose response to EM treatment. The colony formation assay also showed a log-dose response suppression of OVCA cell clonogenicity after treatment with EM. CONCLUSIONS: EM may be a promising candidate in the clinical treatment of patients with OVCA. The lower toxicity and MAP specific action of EM is a novel chemotherapy for OVCA. PMID- 10375111 TI - Epithelial ovarian carcinoma in the reproductive age group. AB - BACKGROUND: A retrospective review of women age < or = 40 years with epithelial ovarian carcinoma was undertaken to determine whether patient age and tumor grade are independent prognostic factors for survival, to investigate the survival rate for young women with ovarian carcinoma, and to characterize these young women in terms of reproductive capability. METHODS: The tumor registry of the Massachusetts General Hospital was used to identify cases of ovarian carcinoma diagnosed between January 1980 and July 1996. Patient records and pathology were reviewed. Survival rates were calculated by the Kaplan-Meier method and Cox proportional hazards models were used to determine the independent effect of each variable on survival. RESULTS: Ninety-two tumors epithelial tumors were identified with 46 (50%) classified as borderline. In the univariate analysis, stage (P < 0.001), grade (P < 0.001), residual disease (< or = 2 cm vs. > 2 cm, P < 0.001), and age (< 30 years vs. 31-40 years; P = 0.019) were found to be significant prognostic factors for survival. However, in the multivariate analysis only tumor grade (with borderline tumors assigned a grade of 0) and stage were significant predictors of survival (P < 0.01 for both). The 5-year survival rate for carcinoma patients with advanced disease was 22.9%. Patients with borderline tumors were more likely be diagnosed during an evaluation for infertility and were more likely to have successful live births after carcinoma treatment. CONCLUSIONS: Young women with advanced epithelial carcinoma have a 5 year survival rate similar to that quoted in the literature, despite the use of more aggressive chemotherapeutic regimens. Patients with borderline tumors of any stage have an excellent prognosis for preserving fertility options. PMID- 10375112 TI - The radial distance of extraprostatic extension of prostate carcinoma: implications for prostate brachytherapy. AB - BACKGROUND: Extraprostatic extension (EPE) is an unfavorable prognostic factor in patients with prostate carcinoma. Prior studies have reported the linear extent of EPE measured circumferentially along the edge of the prostate. In this study, the authors defined and evaluated a novel measure of EPE in a large series of radical prostatectomy specimens. These results have important clinical implications in the management of localized prostate carcinoma by brachytherapy and other modalities. METHODS: The authors reviewed the preoperative records and biopsy findings from 376 patients who underwent radical retropubic prostatectomy between September 1991 and June 1993. Whole mount radical prostatectomy specimens were examined, and the location of EPE for each specimen was recorded. The radial EPE distance was measured perpendicular to the edge of the prostate. For specimens with multiple EPE sites, the maximum radial EPE distance was recorded. Established eligibility criteria for prostate brachytherapy were evaluated using these results, with emphasis placed on achieving adequate radiation dose coverage 3-5 mm beyond the capsule or the edge of the prostate. RESULTS: EPE was identified in 105 of 376 specimens (28%) at 248 sites. The radial EPE distance in these specimens had a mean of 0.8 mm (range, 0.04-4.4 mm) and a median of 0.5 mm. Of these 105 patients, the median and mean preoperative prostate specific antigen (PSA) concentrations were 11.8 ng/mL and 17.9 ng/mL, respectively. The mean and range of the Gleason score and prostate volume for all specimens were 6.3 (range, 3-9) and 39 cc (range, 8-294 cc), respectively. In 107 patients who met the selection criteria for prostate brachytherapy eligibility of a PSA level < 10 ng/mL, Gleason score < 7, and gland volume < 60 cc, the maximum and mean radial EPE distances were 0.6 mm and 0.03 mm, respectively. CONCLUSIONS: The radial distance of EPE is an important measure that influences treatment strategies for patients with localized prostate carcinoma. Currently described criteria for the treatment of early stage prostate carcinoma by brachytherapy alone appear satisfactory to ensure effective radiation dose coverage of EPE of prostate tumors. Treating the prostate with a 3-5 mm margin by brachytherapy would encompass all known tumor in approximately 99% of the specimens examined in this study. PMID- 10375113 TI - Tumor size predicts the survival of patients with pathologic stage T2 bladder carcinoma: a critical evaluation of the depth of muscle invasion. AB - BACKGROUND: Accurate examination of radical cystectomy specimens is critical for stratifying patients into prognostically important groups and determining the need for adjuvant treatment. Evidence has accumulated that cancers invading the superficial muscle wall (T2a) behave similarly to those invading the deep muscle wall (T2b). Quantitative analysis of the depth of invasion in relation to patient outcome is needed. METHODS: The authors systematically evaluated the depth of invasion by micrometer measurement and its relation to the survival of 64 patients with bladder carcinoma pathologic classification as pT2 who had long term follow-up after radical cystectomy. Numerous clinical and pathologic variables were analyzed with univariate and multivariate Cox proportional hazards models. The mean age of patients was 64 years, and their mean follow-up was 8.3 years. RESULTS: There was no significant difference in clinical outcome between patients with T2a carcinoma and those with T2b. Lymph node metastasis and tumor size were each significantly associated with distant metastasis free and cancer specific survival. Ten-year distant metastasis free and cancer specific survival were 100% and 94%, respectively, for patients with tumors <3 cm (P = 0.006) and 68% and 73%, respectively, for patients with tumors > or = 3 cm (P = 0.005). After adjustment for lymph node status, tumor size maintained significance in predicting distant metastasis free survival (risk ratio, 1.5; 95% confidence interval, 1.1-2.0; P = 0.009) and cancer specific survival (risk ratio, 1.5; 95% confidence interval, 1.1-1.9; P = 0.01). Age was associated with recurrence free survival and all-cause survival. None of the other variables, including gender, vascular invasion, presence of carcinoma in situ, pathologic classification (T2a vs. T2b), depth of invasion, depth of muscle invasion, ratio of depth of invasion to bladder wall thickness, and percentage of muscle wall invasion, were significantly associated with patient outcome. CONCLUSIONS: The findings of this study indicate that the subclassification of T2 bladder carcinoma by depth of muscle invasion is of no prognostic value; conversely, tumor size, an easily measured factor, is predictive of distant metastasis free and cancer specific survival. PMID- 10375114 TI - B-lineage lymphoblastic lymphoma is a clinicopathologic entity distinct from other histologically similar aggressive lymphomas with blastic morphology. AB - BACKGROUND: The authors present clinical, histopathologic, and immunophenotypic data regarding B-lineage lymphoblastic lymphoma (B-LBL), a rare entity that has not been extensively studied. To emphasize some of its unique clinical characteristics, the authors compare B-LBL with a group of histologically similar, very aggressive lymphomas, T-lineage lymphoblastic lymphoma (T-LBL) and the blastoid variant of mantle cell lymphoma (BVMCL); all were evaluated concurrently. METHODS: Clinical data were obtained on 29 patients with very aggressive lymphomas (12 B-LBLs, 10 T-LBLs, and 7 BVMCLs) from whom paraffin embedded material was available. The diagnoses were confirmed on review of the hematoxylin and eosin-stained slides and the immunophenotype data. RESULTS: The mean age of patients with B-LBL was 39 years. Patients presented with both lymph node and extranodal disease, although involvement of the mediastinum and bone marrow was infrequent. Four were Stage I, 3 were Stage II, 2 were Stage III, and 3 were Stage IV. B-LBL patients were treated primarily with cyclophosphamide, hydroxydaunomycin, vincristine, and prednisone (CHOP), and one patient underwent allogeneic bone marrow transplantation. The mean follow-up time was 30 months. Seven of 11 had no evidence of disease at 48 months, whereas 4 patients were dead of disease at 5.6 months. The overall median survival was 24 months. The clinical characteristics of B-LBL patients differed significantly from those of T-LBL patients; there was more frequent bone marrow and mediastinal involvement in T LBL cases (P = 0.03 and 0.04, respectively). T-LBL patients were also less likely to achieve a complete remission than B-LBL patients (P = 0.02). The mean age of BVMCL patients significantly exceeded that of B-LBL patients (P = 0.03). CONCLUSIONS: The authors believe that the distinction of B-LBL from its histologic mimics, T-LBL and BVMCL, has important clinical implications. Patients with B-LBL present differently from those with the other very aggressive lymphomas studied, and they achieve complete remissions more often than T-LBL patients. PMID- 10375115 TI - Transient monosomy 7: a case series in children and review of the literature. AB - BACKGROUND: Monosomy 7 and deletions of the long arm of chromosome 7 [del (7q)] are recurrent, nonrandom chromosomal abnormalities associated with both de novo and therapy-related myelodysplastic syndromes (MDS). The overall prognosis for children and adults with these chromosomal abnormalities is poor. In the current report, the authors present five children with MDS associated with monosomy 7/del(7q) who achieved spontaneous hematologic disease remission as well as a review of the literature. METHODS: Five children with either de novo or treatment related MDS who achieved spontaneous hematologic disease remission are presented. Relevant clinical, cytogenetic, and fluorescent in situ hybridization data are included. RESULTS: All patients were boys. Three had de novo MDS whereas two others previously had received chemotherapy for another malignancy. Four patients achieved spontaneous and durable hematologic disease remission that was associated with cytogenetic disease remission in all three patients tested. The fifth patient developed a disease recurrence and died with evidence of clonal evolution after a long interval of hematologic and cytogenetic remission. CONCLUSIONS: A subset of children who develop MDS associated with monosomy 7 or del(7q) achieve spontaneous hematologic and cytogenetic improvement. Although this appears to be uncommon, further data are needed to determine the percentage of patients who improve without therapy and to define clinical characteristics that may predict this clinical outcome. These findings suggest that monosomy 7/del(7q) is insufficient to produce full leukemic transformation. PMID- 10375116 TI - Deletion analysis of the adenomatous polyposis coli and PTCH gene loci in patients with sporadic and nevoid basal cell carcinoma syndrome-associated medulloblastoma. AB - BACKGROUND: Medulloblastomas can occur sporadically or may be associated with hereditary tumor syndromes including familial adenomatous polyposis (FAP) and nevoid basal cell carcinoma syndrome (NBCCS). METHODS: The authors performed a retrospective analysis for allelic deletion of the adenomatous polyposis coli (APC) and PTCH gene loci using paraffin embedded medulloblastoma specimens from patients who were admitted to Children's National Medical Center in Washington, DC, between 1982 and 1997. Thirty-five cases from which tumor and normal tissue could be procured were analyzed. Two of the analyzed cases had a positive family and personal history for NBCCS; in both cases the histology of the medulloblastoma revealed a desmoplastic phenotype. Thirty-three cases were not known to be associated with hereditary disease; 2 of those cases revealed desmoplastic and 31 cases revealed nondesmoplastic "classic" medulloblastoma histology. RESULTS: Although medulloblastoma tumorigenesis has been associated strongly with FAP associated with APC germline mutation, none of the 22 informative sporadic cases revealed loss of heterozygosity of the APC gene locus. PTCH gene deletion was detected in the tumors of both patients with NBCCS. In contrast, only 1 of 33 sporadic medulloblastomas revealed PTCH gene deletion. The sporadic case with PTCH gene deletion did not demonstrate the desmoplastic phenotype. CONCLUSIONS: In conjunction with previous studies, the data from the current study confirm that allelic deletion occurs in NBCCS-associated medulloblastomas, consistent with the role of PTCH as a tumor suppressor gene. However, in sporadic medulloblastomas, allelic deletion of PTCH is an infrequent event. Morphologic examination in conjunction with genetic analysis of PTCH gene deletion in medulloblastoma tissue may prove to be a quick and efficient test with which to screen for NBCCS in patients with medulloblastomas. Although medulloblastoma is a component of Turcot syndrome with demonstrated APC mutations, APC gene deletions appear to be absent or very uncommon in patients with sporadic and NBCCS-associated medulloblastomas. PMID- 10375117 TI - Expression of ras, c-myc, and p53 proteins in cervical intraepithelial neoplasia. PMID- 10375118 TI - Interferon-alpha-associated focal segmental glomerulosclerosis with massive proteinuria in patients with chronic myeloid leukemia following high dose chemotherapy. PMID- 10375119 TI - Recent chromatographic and electrophoretic enantioseparations of cardiovascular drugs. AB - The chromatographic and electrophoretic enantiomeric separation and analysis of several clinically used cardiovascular drugs have been reviewed. Several examples of recently reported applications of enantioselective analysis and various cardiovascular agents are presented. PMID- 10375120 TI - The assay of the catecholamine content of small volumes of human plasma. AB - Plasma catecholamines are routinely measured using high-performance liquid chromatography (HPLC) with electrochemical detection. Most of the present assays require sample volumes of at least 500 microL and are complex and labour intensive procedures, or require large capital investment to reduce the sample size. This paper describes a liquid/liquid plasma catecholamine extraction procedure, HPLC separation and electrochemical detection method which is simple, sensitive and reproducible. The resting catecholamine concentration of 50 microL adult human plasma can be assayed using standard electrochemical detection. The limits of detection were 0.1 fmol injected onto the column for each catecholamine. This method allows the routine assay of plasma catecholamine concentrations within the normal adult ranges in both 500 and 50 microL samples. The within assay coefficient of variation (CV) for noradrenaline (NA) was 1.2% in 500 microL plasma and 1.9% for 50 microL plasma, corresponding values for adrenaline (A) were 8.5 and 6.6%. The between assay CVs were 3.9 and 7.8% for NA, and 9.9 and 5.7% for A. PMID- 10375121 TI - Analysis of morphine and morphine-3beta-D glucuronide in human urine by capillary zone electrophoresis with minimal sample pretreatment. AB - The presence of two of the metabolites of heroin, free morphine and morphine 3 beta-D-glucuronide (MO3G) in acidified urine samples was simultaneously determined by capillary zone electrophoresis (CZE). In a run buffer containing 50 mM sodium borate and 250 mM boric acid (pH 8.6), free morphine migrates before a group of neutral compounds (peak N) in urine, which move with the velocity of electro-osmotic flow. In contrast, the glucuronidated form is negatively charged and migrates behind peak N. Both analytes can be precisely identified within their respective analytical window by their migration time with respect to peak N. The on-line multi-wavelengths scanning of the peak permits further confirmation. The detection sensitivity of both analytes was increased three-four fold if the samples were introduced with electro-injection as compared with hydrodynamic injection. Limits of detection (LOD) of free and conjugated morphine using electro-injection were 200 ng/mL and 500 ng/mL, respectively, determined at a 3:1 signal to noise ratio. A dramatic increase of free morphine was observed after acid hydrolysis of the urine concomitantly with the decrease of the glucuronidated form. We conclude that CZE is a rapid, simple, sensitive and useful screening technique for detection of heroin metabolites in the urine. PMID- 10375122 TI - Biodegradation of pentachlorophenol (PCP) by white rot fungal strains screened from local sources and its estimation by high-performance liquid chromatography. AB - White rot fungal strains screened from local sources (wood trunks and from effluents of pulp and paper industry) were tested for their ability to biodegrade polymeric compounds, viz. polymeric dyes (crystal violet and brilliant green) and chlorinated phenol (pentachlorophenol). Two of the most promising strains showing maximum degradation of polymeric dyes were selected to study the biodegradation potential and pattern of biodegradation of pentachlorophenol (PCP), a commonly used leather preservative and a potential carcinogen. PCP was quantitatively estimated and analysed by high-performance liquid chromatography (HPLC). Conditions were optimized for the measurement of PCP on HPLC, which were: mobile phase, 60% acetonitrile and 40% water; flow rate, 1 mL/ min; column, mu Bondapack C18 RP and UV detector at 238 nm. One of the white rot fungal strains isolated from wood trunk showed a maximum 68% biodegradation of PCP in liquid-buffered medium in 16 days. The biodegradation pattern of PCP followed a pseudo-first order kinetics. Studies on enhancement of biodegradation of polymeric dyes and PCP showed that the kinetics of biodegradation is greatly improved by the presence of manganese ions, H2O2 and glucose in the medium. This strongly suggests the involvement of peroxidase enzyme machinery of white rot fungus in the biodegradation process of PCP. PMID- 10375123 TI - Determination of the relative impact of chromatographic conditions and solute structures on the retention behaviour of non-ionic surfactants in RP-HPLC. AB - The retention behaviour of non-ionic surfactants containing different hydrophobic moiety and various lengths of polar ethylene oxide chain was studied on RP-8 and RP-18 columns using methanol and acetonitrile as organic modifiers. The relative impact of the chromatographic conditions on the retention strength was determined using stepwise regression analysis. Non-ionic surfactants can be separated according to the character of the hydrophobic moiety, but not according to the length of the polar ethylene oxide chain. Calculations proved that the organic modifier exerted the highest impact on the retention; the effect of the number of carbon atoms in the hydrophobic moiety, column temperature and the length of the hydrophobic ligand on the surface of silica support are of secondary importance. PMID- 10375124 TI - Immobilized iminodiacetic acid (IDA)-type Cu2+ -chelating membrane affinity chromatography for purification of bovine liver catalase. AB - A metal ion chelating membrane medium based on iminodiacetate-substituted modified short cotton cellulose was examined for the purification of bovine liver catalase (BLC). The effect of buffer pH, chelator surface density, initial concentration of crude enzyme and flow rate on BLC binding efficiency to the copper ion chelating membrane adsorbent were examined. Under the chromatographic conditions chosen, 67.7% recovery of BLC was attained with an overall 4.2-fold increase in specific activity in a single step. After performance of BLC purification, the chelating membrane adsorbent can be easily regenerated by imidazole or EDTA buffer with higher reviving effectiveness with the latter. PMID- 10375125 TI - Colchicine poisoning: report of a fatal case with body fluid and post-mortem tissue analysis by high-performance liquid chromatography. AB - A case involving a suicide by the ingestion of colchicine tablets is presented. Liquid chromatography has been used to measure the drug level in blood and in post-mortem tissues of the patient (a 42-year-old man). Plasma concentration 24 h after ingestion was 4.5 ng/mL. On autopsy, the kidney showed the highest concentration (396 ng/g). High concentrations were also found in the liver (347 ng/g) and heart (334 ng/g). Low concentrations were detected in the lung (58 ng/g), muscle (10 ng/g) and brain (5 ng/g). PMID- 10375126 TI - Determination of homocysteine in plasma by liquid chromatography with fluorescence detection. AB - A liquid chromatographic (LC) method to assay homocysteine in plasma at micro molar levels is described. The procedure consists of protein precipitation with trichloroacetic acid and a centrifugation step. The supernatant is then derivatized with a thiol-specific fluorochromophore and LC assayed with fluorescence detection at 385 nm excitation and 515 nm detection. LC analysis is performed by isocratic elution using an acetate buffer (0.05 M, pH 4) as a mobile phase and an alphaBond column. Recoveries based on a five-point standard curve calculation ranged from 96 to 112%. The limits of detection and quantification reached were 0.8 and 1.57 micromol/L, respectively. PMID- 10375127 TI - A novel fully automated method for the measurement of sulphoconjugated catecholamines in urine using the Gilson ASTED-XL sample preparation system and high-performance liquid chromatography. AB - Dopamine is produced in the kidney where it causes sodium excretion. Dopamine sulphate is deconjugated in vivo, and may be a physiological reservoir for this active renal dopamine. To investigate the role of dopamine and dopamine sulphate in sodium homeostasis we have developed a fully automated HPLC assay for free, total and sulphoconjugated dopamine. Using the Gilson ASTED-XL sample preparation unit, with temperature controlled racks, urinary free and total dopamine are measured pre-and post-incubation with arylsulphatase. Dopamine sulphate is calculated from the difference between free and total measurements. Acidified 24 h urines are processed automatically. Free dopamine assay follows buffering to pH 7.0, addition of internal standard, addition of EDTA to stabilize free catecholamines at neutral pH, and incubation at 37 degrees C for 30 min. This mixture is trace enriched on a HEMA-SB TEC prior to ion-paired HPLC with amperometric detection. To measure total dopamine the entire process is automatically repeated with addition of arylsulphatase (400 mU/mL urine) at the beginning of the 37 degrees C incubation. The working range of the assay is up to 7 micromol/L total dopamine. Within-and between-run imprecision for dopamine sulphate is less than 3 and 7% respectively. Median dopamine sulphate excretion in 12 normotensive subjects was 4.3 micromol/24 h. PMID- 10375128 TI - A simple chiral high-performance liquid chromatographic method to study the enantiomer-differentiating action of microorganisms. An assay with DL-tryptophan. AB - To investigate the 'enantiomer-differentiating' action of the microorganisms colonizing a phosphate-buffered DL-tryptophan solution, a novel chiral high performance liquid chromatographic (HPLC) arrangement was developed and established. As the HPLC stationary phase, bovine serum albumin (BSA) bonded silica gel was used. In the function of the mobile phase, phosphate-buffered DL tryptophan solution was applied. The composition of the eluate was monitored by an HPLC spectrophotometric detector. After injecting the assayed sample into the eluent stream, the content of each tryptophan enantiomer was evaluated either from the positive or negative responses of the HPLC detector. The validity and the performance of this novel approach were confirmed by applying another chiral HPLC device working with human serum albumin (HSA) bonded silica gel as the stationary phase and with 1-propanol containing phosphate buffer as the eluent. PMID- 10375129 TI - Study of enzyme-catalysed polymerization of lactic acid by HPLC and MALDI-MS. AB - The enzyme-catalysed polymerization of lactic acid was tested by HPLC and MALDI MS methods. A chiral HPLC technique was applied to the investigation of the enantio-selectivity of the reaction. The polymer compounds formed were identified by MALDI-MS. PMID- 10375130 TI - Pulsed high electric field causes 'all or nothing' membrane damage in Listeria monocytogenes and Salmonella typhimurium, but membrane H+-ATPase is not a primary target. AB - Salmonella typhimurium (CRA 1005) was more sensitive than Listeria monocytogenes (NCTC 11994) to pulsed high electric field (PHEF) treatment in distilled water (10, 15 and 20 kV/cm), 10 mM Tris-maleate buffer, pH 7.4 (15 kV/cm) and model beef broth (0.75%, w/v; 15 kV/cm). Sublethal injury could not be detected using a selective medium plating technique, indicating that bacterial inactivation by PHEF may be an 'all or nothing' event. PHEF-induced membrane permeabilisation resulted in an increase in the leakage of UV-absorbing material from the bacteria (UV-leakage) and a decreased ability of L. monocytogenes to maintain a pH gradient. A lack of correlation between the inhibition of H+-ATPase activity and PHEF treatment, cell viability or UV-leakage indicates that this enzyme is probably not a primary site of bacterial inactivation despite its role in the maintenance of internal pH. PMID- 10375131 TI - Fermentation of low-salt miso as affected by supplementation with ethanol. AB - Steam-cooked soybeans and rice koji were combined (1:1, w/w), mixed with 5% (w/w) NaCl and ground into a fine paste. Samples (30 g) were deposited in nylon/polyethylene plastic bags and supplemented with 10 ml of aqueous ethanol solutions to give concentrations of 0, 2.5, 5, 7.5, 10, 15, 20, and 25% ethanol. Mixtures were homogenized, sealed, and incubated at 28 degrees C for eight weeks. Mold populations were less than 3 log10 CFU/g in all miso products after four weeks of fermentation. Yeast populations increased to 6.1 log10 CFU/g in the control (0% added ethanol) during the first week of fermentation and remained stable throughout the eight-week fermentation period. Yeasts were not detected in products containing 5-25% ethanol. Populations of lactic acid bacteria (LAB) increased to 6 log10 CFU/g after one week of fermentation in products containing 0 and 2.5% ethanol. However, after eight weeks of fermentation, LAB populations in all products were less than 4 log10 CFU/g. Rapid decreases in pH occurred only in products supplemented with 0 or 2.5% ethanol. Percentages of soluble protein in miso products containing various ethanol concentrations during the eight-week fermentation period revealed that protease activity was still active or not greatly inhibited in products supplemented with less than 10% ethanol. In comparison, koji enzymes were comparatively less affected by ethanol than were populations of molds, yeasts, and LAB. Total soluble carbohydrate and glucose contents were higher in products supplemented with 5, 7.5 and 10% ethanol than in other products. Discoloration (browning) during fermentation occurred most rapidly in products supplemented with 5 or 7.5% ethanol. Sensory evaluation of the low-salt (5%) product supplemented with 7.5% ethanol and fermented for eight weeks revealed normal or enhanced flavor ratings compared to ratings for a commercial product. PMID- 10375132 TI - Effect of bacteriocin-producing lactobacilli on the survival of Escherichia coli and Listeria in a dynamic model of the stomach and the small intestine. AB - The survival of Lactobacillus curvatus LTH 1174 (bac ) and (bac ) in combination with Escherichia coli LTH 1600 or Listeria innocua DSM20649 during transit through a dynamic model of the human stomach and small intestine (GIT model) was studied. Furthermore, we determined the digestion of curvacin A during gastro intestinal transit and the effect of this bacteriocin on microbial survival. Lb. curvatus is rapidly killed in the gastric compartment at pH < 2.0, and less than 0.01% of the cells delivered to the small intestinal compartments were recovered from the ileal compartment of the model. Meat exerted a protective effect against the lethal action of bile against Lb. curvatus. The sensitivity of E. coli to acid depended on the aeration of the preculture and decreased in the order anaerobic > strongly agitated > agitated. Lactic acid and curvacin A enhanced the lethal effect of low pH on E. coli. Accordingly, cells from strongly agitated cultures were killed faster in the gastric compartment of the GIT model than those from agitated cultures, and inactivation was accelerated in the presence of curvacin A. E. coli tolerated the bile concentrations prevailing in the small intestinal compartments of the model. The survival of Listeria innocua in the GIT model was comparable to that of Lb. curvatus. The curvacin A produced by Lb. curvatus LTH1174 (bac+) killed > 90% of the L. innocua within 10 min after mixing of the cultures. Curvacin A was not degraded in the the gastric compartment, and could be detected in the ileal compartment during the first 180 min upon addition of the meal. PMID- 10375133 TI - Resistance of Escherichia coli and Salmonella against nisin and curvacin A. AB - We have determined the effects of the following factors on the resistance of Gram negative bacteria against nisin and curvacin A: (i) chemotype of the lipopolysaccharide (LPS), (ii) addition of agents permeabilizing the outer membrane, (iii) the fatty acid supply of the growth medium, and (iv) the adaptation to acid and salt stress. Bacteriocin activity was determined against growing and resting cells as well as protoplasts. All smooth strains of Escherichia coli and Salmonella enterica serovar Typhimurium were highly resistant towards the bacteriocins, whereas mutants that possess the core of the LPS, but not the O antigen, as well as deep rough LPS mutants were sensitive. Antibiotics with outer membrane permeabilizing activity, polymyxin B and polymyxin B nonapeptide, increased the sensitivity of smooth E. coli towards nisin, but not that of deep rough mutants. Incorporation of 1 g l(-1) of either oleic acid or linoleic acid to the growth media greatly increased the susceptibility of E. coli LTH1600 and LTH4346 towards bacteriocins. Both strains of E. coli were sensitive to nisin and curvacin A at a pH of less than 5.5 and more than 3% (w/v) NaCl. Adaptation to sublethal pH or higher NaCl concentrations (pH 4.54 and 5.35 or 4.5% (w/v) NaCl) provided only limited protection against the bacteriocidal activity of nisin and curvacin A. Adaptation to 4.5% (w/v) NaCl did not result in cross protection to bacteriocin activity at pH 4.4, but rendered the cells more sensitive towards bacteriocins. PMID- 10375134 TI - Survival of a probiotic, Lactobacillus casei strain Shirota, in the gastrointestinal tract: selective isolation from faeces and identification using monoclonal antibodies. AB - Lactobacillus casei strain Shirota (LCS) is a probiotic bacterium used in the production of fermented milk products and lactic acid bacteria preparations. To investigate the survival of LCS in the gastrointestinal tract, we have developed a selective medium and specific monoclonal antibodies to isolate and identify this strain. Selective LLV agar medium was prepared by modifying LBS medium, a selective medium for lactobacilli, through the replacement of glucose with lactitol as a carbon source and vancomycin as a selective antibiotic. Culture in LLV agar medium followed by ELISA using monoclonal antibodies specific for LCS was able to detect the organism in faeces. Using this method, we studied the faecal recovery of LCS in individuals who drank 125 ml of fermented milk which contained 10(10) live LCS for 3 days. The mean recovery was about 10(7) live bacteria per gram of faeces, indicating that LCS survived transit through the gastrointestinal tract after ingestion of the fermented milk. PMID- 10375135 TI - A survey of the microbiological quality of bottled water sold in the UK and changes occurring during storage. AB - Eight brands of domestic and imported bottled water were microbiologically analysed within three hours of purchase at a local supermarket. Viable numbers of microorganisms were estimated on Plate Count Agar (PCA) and PCA diluted to quarter and tenth strengths (1/4 PCA and 1/10 PCA) and incubated at temperatures of 10, 15, 25 and 37 degrees C. Plate count agar diluted to 1/4 and 1/10 incubated at 25 degrees C yielded the highest initial counts, up to 10(4) cfu ml( 1). Pseudomonas spp. was the predominant species. After 6 months of storage at room temperature (18-25 degrees C), few quantitative and qualitative differences were found in the microflora. PMID- 10375136 TI - Detection of hepatitis A virus in shellfish by nested reverse transcription-PCR. AB - A method for the detection of HAV in shellfish, based on the use of guanidinium isothiocyanate-containing solution for RNA extraction and purification steps, followed by nested PCR, is hereby proposed. Tests were carried out on mollusc samples spiked with HAV strain FG. Results showed that in samples subjected only to one round of PCR it was possible to detect HAV at concentrations of 10(3) 10(4) TCID50/10 g of mollusc. The use of the nested PCR renders the system more sensitive and specific enabling the identification of HAV concentrations as low as 1 TCID50/10 g of mollusc. Furthermore thus method, in addition to allowing the avoidance of confirming tests, such as hybridization, proved to be inexpensive and simple to perform. PMID- 10375137 TI - Factors affecting the efficiency of pooled sample digestion for the recovery of Trichinella spiralis from muscle tissue. AB - Inspection for Trichinella spiralis in pork, horse and game meats is an important part of veterinary public health programmes in many countries. Variations of the pooled sample digestion test are the most widely used methods of inspection for this parasite. In this study, several aspects of the test, including sample preparation, reagent concentration and sample processing were examined for effect on test efficiency. Current methods using sample blending were equivalent or superior to sample grinding. Increasing the concentration of pepsin improved overall digestion slightly. The most critical factor affecting parasite recovery was sieve size. Currently used methods that employ a #80 (180 microm mesh) sieve limit the recovery of motile or dead parasites and therefore might decrease test sensitivity. It is recommended that a #45 (355 microm mesh) sieve be used to ensure optimal recovery of larvae. PMID- 10375138 TI - Combating stigma by providing treatment. PMID- 10375139 TI - Estimating net effects and costs of service options for persons with serious mental illness. PMID- 10375140 TI - Emergency intervention for acute traumatic stress. PMID- 10375141 TI - The importance of psychotherapy in remission and relapse. PMID- 10375142 TI - Web sites worth watching. PMID- 10375144 TI - Satisfaction with managed care among persons with bipolar disorder. PMID- 10375143 TI - Immune function, mood, and perceived burden among caregivers participating in a psychoeducational intervention. PMID- 10375145 TI - A confusion of tongues: competence, insanity, psychiatry, and the law. AB - Psychiatrists share with the public some confusion and uncertainty about two highly visible forensic psychiatric examinations: competence to stand trial and criminal responsibility (insanity). The author reviews the content and context of these examinations, examines legal issues that define and underlie them, and clarifies commonly encountered areas of ambiguity and misunderstanding. The competence examination, which assesses a defendant's ability to participate in the trial process, focuses on the present state of the defendant's mental capacities. Two standards generally used are whether the defendant has a rational and factual understanding of the charges and penalties and has the ability to cooperate with the defense attorney. The examination for insanity is one of the most challenging and comprehensive in forensic psychiatry. The criteria in general address the defendant's awareness of the fact that the act was illegal, wrong, or a crime. Additional criteria address the defendant's ability to control behavior. PMID- 10375146 TI - Mental health issues for Asian Americans. AB - One of fastest-growing population groups in recent decades, Asian Americans represent a vastly diversified and rich mixture of cultures, languages, beliefs, and practices, many of which differ widely from those of European Americans. As immigrants, Asian Americans have experienced and continue to experience various emotional and behavioral problems. However, they tend to underuse existing services except those that are culturally appropriate and linguistically compatible. Misdiagnosis frequently occurs, and the existence of culture-bound syndromes points to a lack of precise correspondence between indigenous labels and established diagnostic categories. Due to Asian traditions of viewing the body and mind as unitary rather than dualistic, patients tend to focus more on physical discomforts than emotional symptoms, leading to an overrepresentation of somatic complaints. Traditional practices and healing methods are frequently used to alleviate distress both before and after patients and their family members approach the conventional mental health care system. Help seeking typically is a family venture. Asian patients respond well to highly structured therapeutic interventions such as those used in behavioral, cognitive, and interpersonal models. When applying pharmacotherapy, clinicians should pay attention to Asians' unique responses to psychotropics, especially in regard to dosage requirements and side effects. Research in this area as well as on other important issues is in the early stage of development. PMID- 10375147 TI - Case managers' and clients' perspectives on a representative payee program. AB - OBJECTIVE: Clients with severe and persistent mental illnesses often require a representative payee to help manage benefit funds. This study compared the perceptions of clients and clinical case managers about the benefits of and problems with the representative payee relationship. METHODS: Fifty-four clients receiving assertive community treatment completed an interview that assessed satisfaction with their experience of having a representative payee and the resulting impact on their substance use, budgeting, and housing. The clients' clinical case managers completed a similar questionnaire. Analyses examined associations between providers' and clients' responses and clients' gender, race, diagnosis, previous experience with a representative payee, and duration of the current representative payeeship. RESULTS: Clients and case managers recognized benefits of the representative payeeship in the areas of housing, substance use, and budgeting. Although little evidence was found that the payeeship pervasively interfered with the therapeutic relationship, 44 percent of case managers reported incidents in which clients verbally abused them over management of their funds. Clients' satisfaction with the representative payeeship was initially low but grew over time. Longer duration of the current payeeship and clients' previous experience with representative payeeship were associated with greater satisfaction and fewer problems. Case managers overestimated clients' initial satisfaction and underestimated their current satisfaction. CONCLUSIONS: Findings suggest that both mental health professionals and clients value the representative payee process as helpful in improving outcomes, although the benefits of the arrangement may be more evident with time and experience. PMID- 10375148 TI - Psychopathy and violent behavior among patients with schizophrenia or schizoaffective disorder. AB - OBJECTIVE: Although a strong association between violence and psychopathy has been demonstrated in nonpsychotic forensic populations, the relationship between psychopathy and violence among patients with schizophrenia has not been thoroughly explored. Patients with and without a history of persistent violent behavior were compared for comorbidity of psychopathy and schizophrenia or schizoaffective disorder. METHODS: Violent and nonviolent patients were identified through reviews of hospital charts and records of arrests and convictions. The Psychopathy Checklist: Screening Version was administered to 51 patients, 26 violent patients and 25 matched nonviolent patients. Analysis of variance was used as the principal statistical method for comparing violent and nonviolent groups. RESULTS: Mean psychopathy scores were higher for violent patients than nonviolent patients. Five of the violent patients (19 percent) had scores exceeding the cutoff for psychopathy, and 13 (50 percent) scored in the possible psychopathic range. All of the nonviolent patients scored below the cutoff for possible psychopathy. Higher psychopathy scores were associated with earlier age of onset of illness and more arrests for both violent and nonviolent offenses. CONCLUSIONS: The comorbidity of schizophrenia and psychopathy was found to be higher among violent patients than among nonviolent patients. Violent patients with schizophrenia who score high on measures of psychopathy may have a personality disorder that precedes the emergence of psychotic symptoms, or they may constitute a previously unclassified subtype of schizophrenia, characterized by early symptoms of conduct disorder symptoms and persistent violent behavior. PMID- 10375149 TI - Development of a consumer survey for behavioral health services. AB - A consumer survey was designed to assess the quality of mental health and substance abuse services and evaluate insurance plans that provide such services. This paper describes the development of the Consumer Assessment of Behavioral Health Services instrument, which began with a review of existing consumer satisfaction surveys and input from several groups working toward development of nationally standardized satisfaction instruments. Consumer focus groups were used to ensure that all the important domains of quality were included, and group members were interviewed to ensure that all items on the instrument were understandable. Results of a pilot test conducted with 160 consumers, 82 enrolled in Medicaid plans and 78 in commercial plans, suggested that the survey was able to distinguish between the two groups in terms of evaluations of their care and insurance plans. Future efforts will focus on further testing of larger, more diverse samples and on developing scoring and reporting formats for the survey that will be useful to consumers and purchasers in choosing behavioral health services and plans. PMID- 10375150 TI - Relationship between maternal church attendance and adolescent mental health and social functioning. AB - OBJECTIVE: This study compared maternal attendance at religious services with standard demographic characteristics such as race, type of religion, and mother's education in terms of their relative association with the behavioral and social functioning of young adolescents. METHODS: The Child Health and Illness Profile- Adolescent Edition and the Children's Depression Inventory were used to screen 445 youths age 11 through 13 who were randomly selected from two public middle schools in Baltimore. Based on the findings, the investigators selected a sample of 143 youths in which approximately two-thirds were at risk of having a psychiatric disorder and the remaining third were unlikely to have a psychiatric disorder. The youths and their mothers were interviewed at home to determine the mothers' frequency of participation in religious services and the youths' self reported health and mental health status and social role functioning. RESULTS: Youths whose mothers attended religious services at least once a week had greater overall satisfaction with their lives, more involvement with their families, and better skills in solving health-related problems and felt greater support from friends compared with youths whose mothers had lower levels of participation in religious services. Maternal attendance at religious services had a strong association with the youths' outcome in overall satisfaction with health and perceived social support from friends, although family income was the strongest predictor of five other aspects of functioning, including academic performance. CONCLUSIONS: Frequent maternal participation in religious services was associated with healthy functioning and well-being in this sample of young adolescents. This association is as important as or more important than associations involving other traditional demographic variables, with the exception of family income. PMID- 10375151 TI - Symptoms predicting inpatient service use among patients with bipolar affective disorder. AB - OBJECTIVE: Symptoms that were risk factors for hospital readmission among psychiatric inpatients diagnosed as having bipolar affective disorder were evaluated. METHODS: Subjects were 100 persons consecutively admitted to a psychiatric inpatient unit at a university-affiliated hospital who met Research Diagnostic Criteria for bipolar I or II disorder or schizoaffective disorder, manic type. Patients were assessed using the Schedule for Affective Disorders and Schizophrenia-Lifetime Version (SADS-L) and the Brief Psychiatric Rating Scale (BPRS) within one week of discharge, and their hospitalization status was documented by monthly phone contacts over a period of 15 months. RESULTS: Twenty four patients (24 percent) were rehospitalized within six months of discharge, and 44 (44 percent) were readmitted within 15 months. Survival analysis using the Cox proportional hazard regression model demonstrated that patients with high scores on a BPRS-derived mania factor were at significantly decreased risk of rehospitalization, whereas those scoring high on a factor consistent with neurovegetative depression were at significantly increased risk. A greater number of previous psychiatric admissions and younger age were also associated with significantly increased risk of rehospitalization. CONCLUSIONS: The findings suggest that patients with bipolar disorder presenting with a depressive episode characterized by prominent neurovegetative features should be treated more aggressively with both pharmacotherapy and intensive outpatient services to reduce the relatively high risk of rehospitalization that appears to be associated with this type of depression. PMID- 10375152 TI - Predicting outcome of inpatient detoxification of substance abusers. AB - OBJECTIVE: Variables that have been identified as predictors of outcome of substance abuse treatment--coping style, addiction severity and addiction-related problems, psychopathology, and treatment motivation--were examined as predictors of outcome of inpatient detoxification. METHODS: A cohort of 175 drug abuse patients consecutively admitted to an inpatient detoxification clinic in the Netherlands were assessed. Baseline data were obtained on psychopathology using the Symptom Checklist-90, on severity and addiction-related problems using the Addiction Severity Index (ASI), on personal coping style using the Utrecht Coping List, on motivation using the CMRS scales, and on sociodemographic background. Positive detoxification outcome was defined as transfer to inpatient rehabilitation treatment. RESULTS: Of the 175 admissions, 81 (46 percent) had a positive outcome, and 94 (54 percent) had a negative outcome. Severe drug use and severe medical problems, as measured by the ASI, were the best predictors of a negative outcome of detoxification. Self-rated suitability of postdetoxification treatment was also a predictor of positive outcome, although to a lesser degree than the ASI variables. Established predictors of residential drug abuse treatment outcome, such as psychopathology, coping style, and sociodemographic variables, did not predict outcome of detoxification. CONCLUSIONS: Caution is necessary when applying results of inpatient treatment outcome studies to inpatient detoxification programs. Different factors may play a role in the outcome of detoxification. To improve the rate at which patients in detoxification programs are transferred to longer-term rehabilitation, more attention should be paid to medical conditions and to the direct consequences of drug use, such as withdrawal symptoms and craving during detoxification. PMID- 10375153 TI - Fidelity to assertive community treatment and client outcomes in the New Hampshire dual disorders study. AB - OBJECTIVE: The study examined the association between fidelity of programs to the assertive community treatment model and client outcomes in dual disorders programs. METHODS: Assertive community treatment programs in the New Hampshire dual disorders study were classified as low-fidelity programs (three programs) or high-fidelity programs (four programs) based on extensive longitudinal process data. The study included 87 clients with a dual diagnosis of severe mental illness and a comorbid substance use disorder. Sixty-one clients were in the high fidelity programs, and 26 were in the low-fidelity programs. Client outcomes were examined in the domains of substance abuse, housing, psychiatric symptoms, functional status, and quality of life, based on interviews conducted every six months for three years. RESULTS: Clients in the high-fidelity assertive community treatment programs showed greater reductions in alcohol and drug use and attained higher rates of remission from substance use disorders than clients in the low fidelity programs. Clients in high-fidelity programs had higher rates of retention in treatment and fewer hospital admissions than those in low-fidelity programs. No differences between groups were found in length of hospital stays and other residential measures, psychiatric symptoms, family and social relations, satisfaction with services, and overall life satisfaction. CONCLUSIONS: Faithful implementation of, and adherence to, the assertive community treatment model for persons with dual disorders was associated with superior outcomes in the substance use domain. The findings underscore the value of measures of model fidelity, and they suggest that local modifications of the assertive community treatment model or failure to comply with it may jeopardize program success. PMID- 10375154 TI - A survey of hospital practices related to pelvic and rectal examinations of psychiatric inpatients. AB - Fifty-three inpatient units in Texas and Massachusetts--15 free-standing psychiatric hospitals and 38 units of general hospitals--responded to a survey to determine current practice in conducting pelvic and rectal examinations of psychiatric patients. Pelvic examinations were never done at 15 of the facilities (28 percent), and rectal examinations were never done at 12 (23 percent). The other facilities did these examinations selectively based on patients' clinical history. Because selective use of these examinations is consistent with recommendations of the American Psychiatric Association and because hospitals that conduct examination selectively have received approval by the Joint Commission on Accreditation of Healthcare Organizations, it appears that selective rather than routine pelvic and rectal examinations are now considered reasonable practice. PMID- 10375155 TI - Health and self-care practices of persons with schizophrenia. AB - Data on the self-care and health-related practices and health status of 22 outpatients with schizophrenia were collected from interviews using a series of health survey instruments and a review of patient records. The patients practiced fewer health-promoting behaviors than nonpsychiatric samples described in the literature and were at risk for premature death primarily due to over-eating and smoking. They were most similar to persons with chronic physical illness in their perception of the locus of control of their health. More than half of the patients had physical conditions requiring medical management. PMID- 10375156 TI - Crowding and aggression on inpatient psychiatric wards. AB - The association between crowding and aggressive behavior among psychiatric inpatients was investigated. Aggressive incidents were documented on two closed psychiatric wards between February 1 and December 15, 1996. A modest correlation between number of patients on the ward and number of aggressive incidents per patient was found. Enlargement of the physical space by the addition of a courtyard to one of the wards midway through the study did not lead to a significant decline in incidents. Possibly, a lack of psychological space-having no privacy or not being able to get sufficient rest-may be more important in triggering aggression than a lack of physical space. PMID- 10375158 TI - A mischaracterization. PMID- 10375157 TI - A tentative model of aggression on inpatient psychiatric wards. AB - Violence in psychiatric hospitals threatens the safety and well-being of patients and staff members. The determinants and correlates of inpatient aggression are not well understood. The authors present an explanatory model of aggressive behavior that attempts to integrate patient, staff, and ward variables. In the proposed model, the patient's psychopathology and distorted cognitions are exacerbated by environmental and communication stressors found on psychiatric wards. The model emphasizes that repeated inpatient aggression may be the result of a vicious circle, whereby a patient's violent behavior is often followed by an increase in stress on the patient caused by environmental or communication factors, heightening the risk of another outburst of violence. PMID- 10375159 TI - Olanzapine and NMS. PMID- 10375160 TI - Pimozide in the treatment of litigious delusions. PMID- 10375161 TI - Antioxidant effects of albumin-bound sulfur in lipid peroxidation of rat liver microsomes. AB - The antioxidant potential of albumin-bound sulfur (SBA) was investigated in rat liver microsomes using lipid peroxidation systems in vitro. Sulfur bound to protein is a reduced metabolite which is produced from cystine by gamma cystathionase. Lipid peroxidation was induced either chemically by ferrous ions and ascorbate or enzymatically by carbon tetrachloride or tert-butyl hydroperoxide as indicated by the increase in thiobarbituric acid reactive substances (TBA-RS) and oxygen consumption. Although the antioxidant effect of SBA was weak on the non-enzymatic lipid peroxidation system, the addition of SBA significantly inhibited TBS-RS formation and oxygen consumption compared with non treated bovine serum alubumin (BSA) in a microsomal lipid peroxidation system induced enzymatically. The sulfur bound to albumin disappeared during incubation with liver microsomes. However, slight differences in the disappearance were observed depending on whether or not lipid peroxidation was induced in the enzymatic systems. In the CCl4-induced lipid peroxidation system, the cytochrome P-450 level was significantly decreased by the addition of SBA. Therefore, in cytochrome P-450 dependent lipid peroxidation system, the potential effects of sulfur bound to albumin are due to an inhibition of cytochrome P-450 rather than by the oxidation itself caused by radical trapping. PMID- 10375162 TI - Enzymatic characterization of human cytosolic sulfotransferases; identification of ST1B2 as a thyroid hormone sulfotransferase. AB - Sulfotransferases (ST) are contained as multiple forms in human tissues with overlapping substrate specificities. To identify a form which contributes to the metabolism of 3, 3',5-triiodothyronine (T3), the functional properties of human STs were compared using recombinant STs, ST1A3, ST1A5, ST1B2, ST1E4 and ST2A3. ST1B2 showed a high affinity (Km 46.2 microM) for T3 sulfation, whereas ST1A3, ST1A5, ST1E4 and ST2A3 showed high affinities to p-nitrophenol (Km 0.4 microM), dopamine (Km 7.1 microM), beta-estradiol (Km 0.3 microM) and dehydroepiandrosterone (Km 3.3 microM), respectively. In Western blotting using antibodies raised against an individual ST, hepatic absolute amounts of these STs were determined. The content of ST1B2 in human liver correlated well with T3 sulfation activities in human liver (r=0.96). These results indicate that ST1B2 is biochemically distinct from other forms of ST, and is involved in the metabolism of T3 in human. In addition, studies of thermal stability and 2,6 dichloro-4-nitrophenol (DCNP) inhibition showed that ST1B2 was thermostable and more DCNP resistant than other forms of ST. Affinities for a co-factor, phosphoadenosine 5'-phosphosulfate, also differed 9-fold among 5 different forms of ST. PMID- 10375163 TI - Comparative studies of new thienothiazine derivatives on heart and smooth muscle preparations of guinea pigs. AB - New thienothiazine derivatives which differ in their side chain on the nitrogen atom of the thienothiazine molecule were studied in guinea pig papillary muscles and terminal ilea using isometric contraction force measurements. Compounds with a heterocyclic ring in their side chain like MS 57 (pyrrolidinylethylcarboxamide side chain), MS 58 (piperidinoethylcarboxamide side chain) and MS 55 (morpholinoethylcarboxamide side chain) had the most potent negative inotropic effect on isolated papillary muscles. The relaxing effect on smooth muscle was more pronounced with compounds carrying an aromatic ring in their side chain like MS 25 (dimethoxyphenylethyl-N-aminopropionyl side chain), MS 24 (dimethoxyphenylethyl-N-methylaminoacetyl side chain) and MS 27 (dimethoxyphenyl N-methylethylaminoethylcarboxamide side chain). Our results show a tissue selectivity of the thienothiazine compounds. PMID- 10375164 TI - A rabbit model for evaluation of chlorpromazine-induced orthostatic hypotension. AB - The present study was conducted to develop an experimental model for evaluation of chlorpromazine-induced orthostatic hypotension in rabbits. In addition, the alpha-adrenoceptor blocking effect of chlorpromazine was investigated in isolated rabbit aorta and saphenous vein in comparison with prazosin. Chlorpromazine (0.1 and 1 mg/kg, i.v.) potentiated significantly a decrease in mean blood pressure at 1 min after the onset of head-up tilt in rabbits anesthetized with urethane alone, urethane+alpha-chloralose or nitrous oxide alone, but not in conscious and morphine+urethane+alpha-chloralose-anesthetized rabbits. There was a negative correlation (r=-0.986, p<0.01) between the extent of chlorpromazine-induced orthostatic hypotension and the amplitude of tilt-induced reflex tachycardia before chlorpromazine treatment. Both prazosin and pentolinium elicited orthostatic hypotension under all four anesthetic conditions. The pA2 value for chlorpromazine to antagonize norepinephrine-induced contraction in aorta was significantly larger than that in saphenous vein, whereas prazosin blocked aortic and venous contractions to a similar extent. These results suggest that a rabbit under an anesthesia which impairs tilt-induced reflex tachycardia may be useful for evaluation of orthostatic hypotension by chlorpromazine. The relatively low potential of chlorpromazine to produce orthostatic hypotension may be partly due to its weak venodilating action. PMID- 10375165 TI - Inhibitory effect of polyoxotungstates on the production of penicillin-binding proteins and beta-lactamase against methicillin-resistant Staphylococcus aureus. AB - In our continuous work on the enhancement of the antibacterial activity of beta lactam antibiotics against the cells of methicillin-resistant Staphylococcus aureus (MRSA) strains by Keggin-structural polyoxotungstates and their lacunary species, Wells-Dawson, double-Keggin, and Keggin-sandwich polyoxotungstates are also found to be synergistic but highly cytotoxic. The coexistence of polylysine or protamine sulphate decreased the synergistic potency of the polyoxotungstates, due to their electrostatic interaction with negatively charged polyoxotungstates. Inductively coupled plasma atomic emission spectrometry (ICP) analysis of the polyoxotungstate-treated cells indicated that the polyoxotungstates uptaken in the cell are preferentially located at the membrane fraction with intact composition. The polyoxotungstates depressed not only the production of PBP2', but also the production of beta-lactamase which hydrolyzes beta-lactam antibiotics on the membrane. This leads to the synergistic effect of polyoxotungstates against the MRSA cells in the coexistence of beta-lactam antibiotics which have high affinities to PBPs 1-4. MRSA cells which were modified to be susceptible to beta-lactam antibiotics during incubation in the presence of polyoxotungstates recovered their resistance to beta-lactam antibiotics when they were subcultured in the absence of the polyoxotungstate. PMID- 10375166 TI - Cell death-inducing activity by gallic acid derivatives. AB - In this study, the cytotoxic activity of gallic acid derivatives (GDs) was studied using some cancer cell lines. Among them, 3,4-methylenedioxyphenyl 3,4,5 trihydroxybenzoate (GD-1) and S-(3,4-methylenedioxyphenyl)-3,4,5-trihydroxy thiobenzoate (GD-3) were found to induce cell death in cancer cell lines with IC50s ranging from 2.9 to 114.4 microM, a concentration comparable with or lower than that of gallic acid. On the other hand, although gallic acid did not show any cytotoxicity against primary cultured rat hepatocytes and human keratinocytes, GD-1 and -3 showed slightly higher sensitivity against such normal cells, when compared with gallic acid. The cell death induced by gallic acid and GD-1 was accompanied by internucleosomal DNA fragmentation characteristic of apoptosis, whereas only smear DNA degradation was detected following GD-3 treatment. When the mechanism by which GD-1 and -3 caused cell death in HL-60RG cells was examined, GD-1 and -3-induced cell death was inhibited by the intracellular Ca2+ chelator, bis-(o-aminophenoxy)-N,N,N,N'-tetraacetic acid acetoxymethyl ester (BAPTA-AM), calmodulin inhibitor, W-7, and the Ca2+/Mg2+ dependent endonuclease inhibitor zinc sulfate. In contrast, catalase, N acetylcysteine (NAC), and ascorbic acid inhibited gallic acid-induced apoptosis in HL-60RG cells, whereas they had no effect on GD-1- and -3-induced cell death. This result suggests that GD-1 and -3 induced cell death in a different manner to gallic acid. In conclusion, esterification of gallic acid with a 3,4 methylenedioxyphenyl group yielded potent agents to treat cancer with a different signaling pathway from gallic acid, although selectivity was lost. PMID- 10375167 TI - Effect of Perilla frutescens extract on nitric oxide production by cultured murine mesangial cells. AB - The effects of a water extract of perilla (Perilla frutescens Britton) leaves on nitric oxide (NO) production by cultured murine mesangial cells were investigated. Perilla extract significantly induced NO production from mesangial cells, which was enormously augmented without cytotoxity by combination with interferon (IFN)-gamma or tumor necrosis factor-alpha. On the other hand, perilla extract suppressed a large amount of NO production induced by IFN-gamma combined with lipopolysaccharide. Northern blot analysis revealed that such effects of perilla extract were dependent on inducible NO synthase mRNA expression. Perilla extract exhibited an inhibitory effect on cytokine-induced mesangial cell proliferation, and this effect was significantly decreased upon combination with N(G)-monomethyl-L-arginine (L-NMMA), a non-specific NO synthase inhibitor, suggesting that perilla extract inhibits mesangial cell proliferation partially through the induction of NO production. Such results indicate that perilla may be a promising agent for the prevention of the progression of glomerulonephritis. PMID- 10375168 TI - DNA profiling of Acorus calamus chemotypes differing in essential oil composition. AB - The phylogenetic relationship of Acorus gramineus and three types of Acorus calamus was analyzed by comparing the 700 bp sequence of a 5S-rRNA gene spacer region. Although there was no intra-specific variation in the essential oil profile of A. gramineus which contained a phenylpropanoid (Z-asarone) as a predominant constituent, A. calamus was classified into two chemotypes: chemotype A in which Z-asarone is a major essential oil constituent and chemotype B which contained sesquiterpenoids predominantly. An intermediate type (M) of these two chemotypes in various ratios was also observable. The NJ tree constructed based on the sequences revealed that A. gramineus was clearly distinguished from any of the chemotypes of A. calamus and that the phylogenetic relationship predicted by the spacer region data correlated well with the essential oil chemotype pattern of A. calamus. PMID- 10375169 TI - Antihyperlipidemic action of Ogi-Keishi-Gomotsu-To-Ka-Kojin against cyclophosphamide-induced hyperlipidemia in rabbits. AB - The effect of Ogi-Keishi-Gomotsu-To-Ka-Kojin (OKGK), a Japanese traditional herbal medicine (Kampo medicine), has been studied in a cyclophosphamide (CPM) induced hyperlipidemia model in fasted rabbits. In this model, the accumulation of chylomicrons and very low density lipoprotein (VLDL) was known to occur as a result of a reduction in lipoprotein lipase (LPL) activity in the heart and heparin-releasable heart LPL. Oral administration of OKGK for 4 weeks was found to reverse the increase in serum triglycerides and cholesterol produced by CPM treatment especially in chyromicrons and VLDL. In addition, OKGK treatment led to a recovery in postheparin plasma LPL activity and heparin-releasable heart LPL activity which were reduced markedly by CPM treatment. We previously reported that OKGK increased LPL activity in postheparin plasma in rats. In this study, we have also found that OKGK improved hyperlipidemia in the CPM-induced hyperlipidemia model in rabbits, mainly due to an increase in heparin-releasable heart LPL activity and postheparin plasma LPL activity. PMID- 10375170 TI - Anticonvulsant activity of paeonimetabolin-I adducts obtained by incubation of paeoniflorin and thiol compounds with Lactobacillus brevis. AB - Seventeen thiopaeonimetabolin-I adducts were obtained as mixtures of diastereoisomers after incubation of paeoniflorin with Lactobacillus brevis in the presence of various thiols. The anticonvulsant activity of the adducts was investigated in mice using the maximal subcutaneous pentylenetetrazol seizure test and sodium valproate (1.5 mmol/kg) as a positive control. Thirteen adducts showed dose-dependent prolongation of latencies of clonic and tonic convulsions. Maximal protection against convulsions was effectively demonstrated by 8-(n hexylthio)paeonimetabolin I (8) and 8-benzoylthiopaeonimetabolin I (18) at doses of 0.125 and 0.25 mmol/kg, respectively, while 100% protection was only achieved at 0.5 mmol/kg of 8-cyclopentylthiopaeonimetabolin I and 8-(p tolylthio)paeonimetabolin I. The principal anticonvulsant activity of the diastereoisomers of 8 and 18 was attributed to their 7S-isomers [ED50 values of 0.09 and 0.12 mmol/kg, and protective indices of 5.0 and 4.0 for 8 (7S) and 18 (7S), respectively], while the 7R counterparts [8 (7R) and 18 (7R)] showed a muscle relaxation effect. PMID- 10375171 TI - Stereoselective pharmacokinetics of RS-8359, a selective and reversible inhibitor of A-type monoamine oxidase, in rats, mice, dogs, and monkeys. AB - RS-8359, (+/-)-4-(4-cyanoanilino)-5,6-dihydro-7-hydroxy-7H-cyclopenta[d]-py rimidine, inhibits, selectively and reversely, A-type monoamine oxidase (MAO-A). In order to clarify the stereoselective metabolism of this drug, plasma concentrations of the [R] and [S]-enantiomers of RS-8359 were determined by chiral column HPLC after oral administration of each enantiomer to rats, mice, dogs, and monkeys. After administration of the [R]-enantiomer, high levels were retained in all animal species studied. On the other hand, when the [S] enantiomer was administered, plasma concentrations decreased rapidly in rats and mice, and extremely rapidly in dogs, while in monkeys, only a trace amount was detected immediately after dosing. Thus, it was found, as a common phenomenon in rats, mice, dogs, and monkeys, that plasma concentrations of the [S]-enantiomer were markedly lower than those of the [R]-enantiomer. Secondly, the [R] enantiomer was observed in plasma after administration of the [S]-enantiomer, and the [S] to [R] chiral inversion rate was estimated from AUC([R] after [S])/AUC([R] after [R]). The percentage was 45.8% in rats, 3.8% in mice, 0.8% in dogs, and 4.2% in monkeys. Further, the [S]-enantiomer was detected in plasma of SD rats dosed with the [R]-enantiomer, suggesting [R] to [S] chiral inversion in rats. These results show marked species differences in the chiral inversion of the cyclopentanol group of RS-8359. A mechanism of chiral inversion is discussed based on experiments using isolated rat hepatocytes. PMID- 10375172 TI - Effect of clarithromycin and other macrolides on the sulfoxidation and 5 hydroxylation of lansoprazole in dogs. AB - The purpose of this study was to evaluate a possible interaction between lansoprazole and clarithromycin as well as other macrolides in dogs. Lansoprazole (30 mg) was orally administered to male beagle dogs, with or without oral pretreatment with 200-mg clarithromycin twice a day for 5 d. The experiments had a randomized cross-over design with a two-week washout period between dosing regimens. Clarithromycin pretreatment for 5 d resulted in a significant increase in the area under the serum lansoprazole concentration-time curve (AUC), whereas the area for a lansoprazole metabolite, lansoprazole sulfone, was significantly reduced, as was the maximum serum concentration (Cmax) of lansoprazole sulfone. When the effects of clarithromycin on the metabolism of lansoprazole were studied using dog liver microsomes, it was found that clarithromycin significantly inhibited the formation of lansoprazole sulfone but not another metabolite, 5 hydroxylansoprazole. These results suggest that co-medication of lansoprazole with clarithromycin may produce a synergistic effect caused by the increased serum levels of lansoprazole of benefit in Helicobacter pylori eradication. PMID- 10375173 TI - Tracheal barrier and the permeability of hydrophilic drugs and dipeptides. AB - The permeability of model hydrophilic compounds with different molecular weights and model dipeptides were examined to characterize the tracheal epithelial barrier in in vitro experiments using excised rabbit trachea. 6 Carboxyfluorescein (6-CF; 376 Da) and fluorescein isothiocyanate (FITC)-dextrans (FDs) with varying molecular weights (4 to 70 kDa) were used as model hydrophilic and macromolecular compounds, and glycyl-D-phenylalanine (Gly-D-Phe) and glycyl-L phenylalanine (Gly-L-Phe) were used as model dipeptides in this experiment. The apparent permeability coefficients (Papp) of 6-CF and FDs with Mw 376 Da to 70 kDa ranged from 2.35x10(-7) to 4.05x10(-10)cm/s and exhibited a good inverse correlation with their molecular weights. The tracheal permeability of 6-CF, FD-4 (4 kDa) and FD-10 (10 kDa) were increased over three fold by 10 mM glycocholate, which is an absorption enhancer. The Papp of Gly-D-Phe was 1.03x10(-6)cm/s and there was no metabolism during tracheal permeation. Gly-L-Phe was immediately degraded in the mucosal fluid and its intact form was not detected in serosal fluid during the 150 min experimental period. These results suggest that absorption of some peptide drugs via the respiratory tract may contribute to their systemic delivery following pulmonary administration by intratracheal insufflation and instillation. PMID- 10375174 TI - In vivo studies on the role of complement in the clearance of liposomes in rats and guinea pigs. AB - The ability of complement (C) system to remove liposomes from blood circulation was examined in vivo using rat and guinea pig as models. Although the liposomes were not degraded in guinea pig serum in vitro, they were degraded remarkably in guinea pig circulation, as assessed by the urinary excretion of [3H]inulin released from liposomes. The suppression of rat C system to 64% normal C hemolytic activity by treating animals with K76COOH agent resulted in a significant decrease in both the uptake of liposomes by liver and the release of [3H]inulin, providing in vivo evidence for C-mediated clearance of liposomes in rats via uptake by macrophages and degradation in blood circulation, respectively. On the other hand, the K76COOH-induced suppression of C (70% normal hemolytic activity) in guinea pigs slightly increased both the hepatic uptake and the release of [3H]inulin. In addition, the hepatic uptake and in vivo degradation in guinea pigs varied in an opposite manner when the animals were preloaded by empty liposomes or when the liposome size and cholesterol content varied. These results suggest there is a difference between the factors involved in liposome degradation and the factors involved in hepatic uptake and also support the likelihood that there is no C-mediated degradation in guinea pigs. PMID- 10375175 TI - Rectal absorption of [Bis (acetato) ammine dichloro (cyclohexylamine) platinum(IV)] (BMS-182751), a new anti-tumor agent, in rats. AB - The rectal absorption of a platinum anti-tumor agent, [bis (acetato) ammine dichloro (cyclohexylamine) platinum(IV)] (BMS-182751), was investigated in rats. BMS-182751 was co-ground with various carriers to improve its poor aqueous solubility. The highest drug dissolution was observed for the co-ground mixture of BMS-182751 and low molecular (LM) gelatin (1:9, w/w), followed by beta cyclodextrin and polyvinylpyrrolidone. The influence of a suppository base or additive on the rectal absorption of BMS-182751 in the drug state of crystalline powder or co-ground mixture was examined in vitro using excised rat rectum. A macrogol base gave much higher BMS-182751 permeation across the rat rectum than that from a Pharmasol base. The addition of sodium caprylate or caprylic acid to the macrogol base markedly enhanced the drug permeation, and a 3% addition of sodium caprylate to the base afforded maximum drug permeation. Two rectal formulations, the co-ground mixture with LM-gelatin plus 3% sodium caprylate in macrogol and the crystalline drug alone plus 3% sodium caprylate in macrogol, as well as an oral aqueous drug suspension, were administered to rats. The Cmax and AUC0-24h values for platinum from the former suppository were 5.1- and 4.1-fold greater than those from the oral suspension, respectively. The values from the latter suppository were almost comparable to those from the suspension. These results suggest that the suppository may provide a promising therapeutic means for cancer treatment using this platinum agent. PMID- 10375177 TI - Cycloprodigiosin hydrochloride obtained from Pseudoalteromonas denitrificans is a potent antimalarial agent. AB - Cycloprodigiosin hydrochloride (cPrG*HCl) is a stable fluorescent red pigment obtained from the marine bacterium Pseudoalteromonas denitrificans. It was found that the compound was incorporated into Plasmodium falciparum cells upon incubation and exhibited a potent antimalarial activity with the concentration required for 50% of the activity being 11 nM, which is stronger than that of chloroquine, a well-known antimalarial agent. The compound did not affect growth rate of mammalian cells. Antimalarial activity of cPrG*HCl was also observed in vivo. These results indicate that cPrG*HCl is a potent antimalarial drug. PMID- 10375176 TI - Studies on interactions between traditional herbal and Western medicines. I. Effects of Sho-seiryu-to on the pharmacokinetics of carbamazepine in rats. AB - The effects of oral co- and pre-administration of Sho-seiryu-to extract powder (TJ-19, 1 g/kg), a widely used Kampo (traditional Chinese herbal) medicine, on the pharmacokinetics of an anti-epileptic drug, carbamazepine (CBZ), and its active metabolite (carbamazepine-10,11-epoxide, CBZ-E) after oral administration of CBZ (50 mg/kg) were examined in male rats. The simultaneous administration of TJ-19 significantly lengthened the time to reach the peak plasma concentration (Tmax), but did not influence the peak plasma concentration, area under the plasma concentration-time curve or terminal elimination half-life (t1/2). Each parameter for CBZ or CBZ-E with a single pretreatment with TJ-19 was not significantly different from that with the vehicle. Tmax and the elimination rate constant for CBZ were significantly increased by 1-week repeated pretreatment with TJ-19, by 83% (p<0.01) and 88% (p<0.001), respectively. t1/2 and the mean residence time from zero to infinity (MRT0-infinity) in the TJ-19 pretreatment group were significantly shortened, by 52 and 34% (p<0.005), respectively. No significant difference in the bound fraction of each drug at two concentrations (1 and 10 microg/ml) was observed between the control and TJ-19 pretreatment groups. These results indicate that simultaneous oral administration of TJ-19 delays the oral absorption of CBZ, while 1-week repeated pretreatment with TJ-19 accelerates the metabolism of CBZ in rats, without affecting the protein binding of CBZ. PMID- 10375178 TI - Different absorption behaviors among steroid hormones due to possible interaction with P-glycoprotein in the rat small intestine. AB - The intestinal absorption of ten steroid hormones was evaluated in the rat small intestine, especially focusing on the interaction with intestinal P-glycoprotein (P-gp). Hydrocortisone, prednisolone, 6alpha-methylprednisolone, and dexamethasone (adrenocortical steroid hormones) all disappeared in a regional dependent manner (duodenum>jejunum>ileum). The decreased rate of disappearance in the lower small intestine seemed to be due to the involvement of absorption barriers like P-gp. In contrast, all sex hormones including progesterone exhibited very high absorbability in the entire small intestine (duodenum=jejunum=ileum), possibly demonstrating the absence of restricted absorption by intestinal P-gp. Progesterone enhanced the rate of disappearance of vinblastine but did not affect 6alpha-methylprednisolone. In the presence of vinblastine and verapamil, on the other hand, the rate of disappearance of 6alpha methylprednisolone increased significantly. It was demonstrated that there was a plural P-gp family, which had different substrate specificities, in the rat intestine and that steroid hormones interacted with them as substrates or inhibitors in a very complex manner. PMID- 10375179 TI - A modified fluorescence polarization immunoassay method incorporating fat emulsion (FE-FPIA) to determine cyclosporin A concentrations in rat skin. AB - We have developed a simple, sensitive and reliable assay procedure for cyclosporin A (CyA), a modified fluorescence polarization immunoassay method incorporating fat emulsion (FE-FPIA), to determine the CyA content in rat skin. The conventional fluorescence polarization immunoassay (FPIA) method for CyA using a commercially available FPIA kit, TDX cyclosporine monoclonal whole blood, was modified. A fat emulsion for intravenous infusion, Intralipos, was incorporated for dissolving the CyA extracted from the skin tissue, and a mixture of MeOH/purified water was used as the sample pretreatment medium instead of the precipitation reagent in the conventional FPIA kit intended for whole blood samples. These modifications enabled us to produce a reliable and the sensitive assay of CyA in skin tissue. The reproducibility (coefficient of variation), detection limit, and assay time for FE-FPIA were below 2%, 25 ng/ml, and about 24 min/24 samples, respectively, and were comparable with those for the whole blood samples determined by the conventional FPIA. Pre-purification of samples required by the HPLC assay is not needed in the FE-FPIA method. The usefulness of the FE FPIA method in evaluating the topical pharmacokinetics of CyA in skin is discussed. PMID- 10375181 TI - A new cytotoxic 2H-pyran compound produced by acid treatment of the fermentation broth of Streptomyces sp. AB - A new cytotoxic substance was isolated following acid treatment of fermented broth of a Streptomyces strain numbered MJ915-WF12. The structure was elucidated by spectroscopic analyses to be 5-dihydro-4-formyl-6-hydroxy-2-hydroxymethyl-6 methyl-2H-pyran. It was thought to be generated by dehydration and ring reformation of the precursor material in cultured broth by acid. PMID- 10375182 TI - Binding of Asp-hemolysin from Aspergillus fumigatus to oxidized low density lipoprotein. AB - Asp-hemolysin is a human low density lipoprotein (LDL) binding protein. We found evidence that Asp-hemolysin binds to oxidized LDL (oxLDL) as well as to LDL in a concentration-dependent manner. This result suggests that Asp-hemolysin is a novel protein, which shares binding abilities to LDL and oxLDL. PMID- 10375180 TI - Effect of a Dynorphin A analog, E2078, on phospholipid membrane properties. AB - The effect of the Dynorphin A analog, E2078, on the physicochemical properties of lipid membranes was investigated. E2078 induces the leakage of calcein, especially out of negatively-charged vesicles. The peptide binds to dipalmitoylphosphatidylglycerol sonicated vesicles according to the Lamgmuir isotherm, with a binding constant of 4.0x10(5) M(-1) and a binding-site number of 0.14 per lipid molecule. The leakage seems to occur at a critical binding-site number of approx. 0.025 per lipid molecule. In addition, E2078 increased the membrane fluidity of the acidic lipid bilayer. In conclusion, E2078 interacts with acidic lipids through electrostatic interactions, inducing leakage and increasing membrane fluidity. PMID- 10375183 TI - Ureteropyeloscopy in the diagnosis of patients with upper tract hematuria: an initial clinical study. AB - BACKGROUND: To study the usefulness and safety of ureteropyeloscopy in the diagnosis of upper tract hematuria of unknown etiology by standard diagnostic methods. METHODS: Fifteen patients with upper tract hematuria of unknown etiology were the subjects of the present study. Prior to ureteropyeloscopy, they underwent standard diagnostic methods, including cystourethroscopy, excretory urography and computed tomography scan. The upper tract (ureter, renal pelvis and calyces) was inspected systematically with a flexible ureteropyeloscope under epidural anesthesia. A biopsy specimen was obtained when neoplasm of a suspicious lesion was seen. Bleeding and hemangiomatous lesions were fulgurated at the time of ureteropyeloscopy. RESULTS: Unilateral gross hematuria was seen in 12 patients. Imaging studies revealed a filling defect in four patients, ureteral stenosis in one patient and nutcracker phenomenon in one patient. Urine cytology was positive in three patients and suspicious in four patients. Results of ureteropyeloscopy were papillary tumor in three patients, whitish encrustation in one patient, redness of the renal pelvis in one patient, bleeding from the renal calyx in two patients, hemangiomatous lesion in one patient, ureteral stenosis in two patients and no abnormalities in five patients. Biopsies were performed in five patients. The pathology results were transitional cell carcinoma in four patients and no abnormality in one patient. Although a ureteral stent catheter was placed in one patient, no serious complications were encountered during or after the procedures. CONCLUSIONS: Ureteropyeloscopy was useful and relatively safe. This endoscopic examination can differentiate insignificant lesions from significant lesions by visual inspection of the lesions, in addition, pathological diagnosis by biopsy specimen can also be performed if deemed necessary. Ureteropyeloscopy is recommended in the diagnosis of upper tract hematuria of unknown etiology. PMID- 10375184 TI - Evaluation of functional bladder capacity in Japanese children. AB - BACKGROUND: Functional bladder capacity is a very important factor in the diagnosis of children with voiding disorders. Because Japanese children are thought to have somewhat smaller functional bladder capacity compared with Western children, the convenient formula (so-called Koff formula) bladder capacity (in ounces) = age (in years)+ 2 is not suitable for use in Japanese children. METHODS: We measured the bladder capacities of 131 Japanese children aged 5-15 years without clinical voiding pattern abnormalities to develop a practical guideline for the prediction of normal bladder capacity for age. RESULTS: An approximate formula relates age and bladder capacity as: bladder capacity (mL) = 25 x (age (years) + 2). CONCLUSIONS: The formula presented is thought to be a useful guide for the diagnosis of small, normal or large bladder capacity and offers information on voiding disorders in Japanese children. PMID- 10375185 TI - Early results of radical prostatectomy and adjuvant endocrine therapy for prostate cancer with or without preoperative androgen deprivation. The Prostate Cancer Study Group. AB - BACKGROUND: The effects of preoperative androgen deprivation were explored in the patients who received radical prostatectomy and subsequent adjuvant endocrine therapy for prostate cancer. METHODS: Stage A2, B or C prostate cancers were randomized to one of two groups: (i) group I (n=90), who received androgen deprivation (leuploride and chlormadinone acetate) for 3 months preoperatively followed by radical prostatectomy and adjuvant endocrine therapy (leuploride only); and (ii) group II (n=86), who underwent the surgery followed by 3 month androgen deprivation and subsequent adjuvant endocrine therapy. The effects of preoperative androgen deprivation on clinical relapse (serum prostate specific antigen (PSA) > 1.98 ng/mL, local recurrence or distant metastasis) and PSA relapse (PSA >0.2ng/mL) were evaluated at 2 years after randomization. RESULTS: There was no significant difference in clinical or PSA relapse-free survival and quality of life measures between the two groups, although relapses occurred significantly more frequently in patients who had more advanced stages, higher pretreatment PSA values or lower histologic differentiation in either group. Subgroup analysis indicated that clinical relapse-free survival in stage C cancer tended to be better in patients with preoperative androgen deprivation than in those patients without it (P< 0.1). CONCLUSIONS: Preoperative androgen deprivation may be beneficial for stage C prostate cancer patients receiving radical prostatectomy and adjuvant endocrine therapy over the 2 year observation period. A longer follow up is needed to clarify the exact extent of benefit in terms of survival and quality of life. PMID- 10375187 TI - Role of protein kinase C in cisplatin nephrotoxicity. AB - BACKGROUND: Cisplatin is widely used in cancer treatment. The major disadvantage of this antitumor agent is its nephrotoxicity. The mechanism of cisplatin-induced nephrotoxicity has not been clarified. Recent evidence suggests protein kinase C (PKC)-related signal transduction pathways may modulate cisplatin-induced cytotoxicity. METHODS: The effect of cisplatin administration on PKC expression in the kidney and the effect of a PKC inhibitor on cisplatin-induced renal impairment were investigated in rats. RESULTS: A single intraperitoneal injection of 8 mg/kg cisplatin induced remarkable damage in the proximal tubules located in the outer medulla, which was associated with impaired renal function, within 48 h. An immunoblotting study revealed marked expression of alpha-PKC in membrane fractions of medullary tubules prepared from cisplatin-treated rats. In addition, pretreatment with the PKC inhibitor (H-7) protected kidneys from cisplatin induced damage. CONCLUSIONS: These findings suggest that the nephrotoxic effects of cisplatin may, in part, be related to PKC activation in the renal tubules. PMID- 10375186 TI - Pattern of progression and survival in hormonally treated metastatic prostate cancer. AB - BACKGROUND: Patients with prostate cancer generally respond to androgen ablation therapy, but progression to androgen-independence is frequently observed. To further evaluate disease progression, the pattern of progression and survival in hormonally treated metastatic prostate cancer was examined. METHODS: One hundred and ninety-three patients with untreated metastatic prostate cancer (TxNxM ) who received endocrine therapy between 1986 and 1995 were included in the present study. The pattern of progression was evaluated in these patients. RESULTS: One hundred and eighteen of the 193 patients (61.1%) had disease progression: 33 had local progression, 73 had distant progression and 12 had distant with local progression. Patients with only local progression had a longer interval to disease progression and longer survival than those with distant progression. The interval from disease progression to death in patients with local progression was longer than in those with distant progression. The patients whose prostate specific antigen (PSA) had not been normalized 3 months after the start of endocrine therapy had a tendency to progression either into the prostate or into distant sites. Patients with extent of disease (EOD) scores of 3 and 4 progress, especially to distant sites, after endocrine treatment. CONCLUSIONS: In untreated metastatic prostate cancer, patients with a poor response of PSA levels and patients with a high volume of bone metastasis (i.e. EOD 3, 4) were in the high risk group for progression, especially to distant sites. Progression into distant sites was a poor prognostic factor for patients with recurrence to endocrine therapy. PMID- 10375188 TI - Electrical activity of the corpus cavernosum in denervated rats. AB - BACKGROUND: We evaluated the electrical activity of the corpus cavernosum after intracavernous papaverine injection in rats that had been denervated experimentally. METHODS: Twenty-four male adult Sprague Dawley rats were divided into three groups: (i) controls (n=8) (ii) unilateral cavernous nerve resection on the right side (n=8); and (iii) bilateral cavernous nerve resection (n=8). Through a suprapubic incision, the urinary bladder was retracted laterally to locate the major pelvic plexus on the lateral surface of the prostate. The major branch of the cavernous nerve, running caudally from the pelvic plexus, was isolated and excised using an operating microscope. Three weeks later, recording of the electrical activity of the corpus cavernosum (EACC) was performed by using a Neuropack-2 EMG unit (Nihon Kohden, Tokyo, Japan) and coencentric needle electrode. Changes in amplitude were evaluated before and after intracavernosal papaverine injection. The results in the flaccid state and after papaverine injection were compared by using the Mann Whitney U-test in all three groups and paired t-test between groups. RESULTS: In the flaccid penis, the mean (+/- SD) amplitude of electrical activity of the corpus cavernosum was 17.42+/-2.05, 12.42+/-1.88, 9.71+/-1.59 and 5.85+/-0.96 microV in control rats, in unilaterally denervated rats (in which the cavernous nerve was intact on the left side), in unilaterally denervated rats in which the cavernous nerve was resected on the right side and in bilaterally denervated rats, respectively. In the flaccid state, EACC is lower in the bilaterally denervated group than in the control and unilaterally nerve-resected groups (P < or = 0.05). The recording of electrical activity of the corpus cavernosum was continued for 20 min after papaverine injection. In the control group and in both groups of unilaterally denervated rats, we observed a significant decrease in the electrical activity of the corpus cavernosum in the first 5 min after papaverine injection (P < or = 0.05). However, no difference was observed in bilaterally denervated rats after injection (P > or = 0.05). CONCLUSIONS: We conclude that electrical activity of the corpus cavernosum continues after unilateral nerve injury in rats. Cross innervation may play a role in penile innervation and corpus cavernosum electromyography shows electrical activity in denervated rats. PMID- 10375189 TI - Psoas abscess associated with iliac vein thrombosis and piriformis and gluteal abscesses. AB - BACKGROUND: A 14-year-old boy was admitted because of lumbago and high fever. METHODS/RESULTS: Computed tomography scans revealed psoas, piriformis and gluteal abscesses as well as right iliac vein thrombus. A right femoral venogram demonstrated compression from the psoas abscess and thrombosis of the common iliac vein. Appropriate surgical drainage, administration of antibiotics and anticoagulant therapy were effective in the present case. CONCLUSIONS: This is the first report of primary psoas abscess associated with vein thrombosis and is also unique in that abscesses were multiple without predisposing diseases or trauma. PMID- 10375190 TI - Non-traumatic renal arteriopelvic fistula. AB - PURPOSE: In the present paper, we report on a 34-year-old female with macroscopic hematuria due to a nontraumatic renal arteriopelvic fistula (APF). The patient initially presented at another hospital with asymptomatic macroscopic hematuria. Following abdominal ultrasonography, computed tomography (CT) and laboratory data, no abnormal findings were seen. Therefore, the patient was referred to Teine Keijinkai Hospital for a more precise evaluation of the urinary tract and vascular abnormality. METHODS/RESULTS: Endoscopically, there was bleeding from the right ureteral orifice, so the patient was admitted for further examination. No abnormal findings were seen on urinary cytology and following an intravenous pyelogram. A selective right lower polar renal arteriogram revealed arterial extravasation directly into the pelvis before the venous phase, so APF of the kidney was diagnosed. The patient had no history of urinary tract trauma, so the APF was thought to be idiopathic. After transcatheter arterial embolization (TAE) with a gelatine sponge, macroscopic and microscopic hematuria disappeared and a low-density area was seen in the middle pole of the right kidney in an abdominal CT scan 4 days after TAE. This was thought to be renal infarction due to TAE. CONCLUSIONS: After discharge, the patient had no further hematuria. PMID- 10375191 TI - Intra-abdominal abscess due to patient non-compliance after construction of an ileal neobladder: case report and review of the literature. AB - PURPOSE: A case report of patient with an intra-abdominal abscess 8 weeks after radical cystectomy and construction of an ileal neobladder is presented. METHODS/RESULTS: The patient was admitted with nausea, vomiting and singultus. A perforation of the neobladder due to overdistension was assumed to be the underlying cause of the intra-abdominal abscess formation as the patient admitted infrequent voiding during the day and no emptying of the neobladder at night. The patient underwent explorative laparotomy and 4200 mL of pus was removed from the abdominal cavity. The patient made an uneventful recovery and was discharged from hospital after 5 weeks. Neobladder function remained stable and the patient was leading a normal life at 24 months follow-up. CONCLUSIONS: The present case demonstrates the need for careful patient selection prior to radical cystectomy with continent urinary diversion. Reduced compliance and mental disabilities of a patient can increase the complication rate. PMID- 10375192 TI - Angiomyofibroblastoma of the female urethra. AB - BACKGROUND: Angiomyofibroblastoma is a relatively recently described rare tumor of the superficial soft tissues. To date, 57 cases of angiomyofibroblastoma of the external genitalia in women have been reported. METHODS/RESULTS: We describe a case of a 24-year-old woman who presented with the urinary stream flowing out in the posterior direction and whose diagnosis was a urethral tumor, angiomyofibroblastoma. CONCLUSIONS: Angiomyofibroblastoma has a potential arising from the female urethra as well as other areas of the external genitalia. PMID- 10375193 TI - Clinical experiences of microsurgical side-to-end epididymovasostomy for epididymal obstruction. AB - BACKGROUND: Some surgical treatments are performed for obstructive azoospermia in urology and good results have been reported. Of 61 azoospermic patients who visited our department of urology, nine were diagnosed as having epididymal obstruction of unknown etiology. METHODS: We describe nine consecutive side-to end epididymovasostomy procedures performed on these patients. These procedures are microsurgical two-layer anastomosis. RESULTS: Of the nine men, five (55.6%) had sperm in the ejaculate postoperatively and, up until publication, the pregnancy rate was 33.3% (three of nine). CONCLUSIONS: These results suggest that reconstruction of the seminal tract should be considered first for obstructive azoospermia. PMID- 10375194 TI - Ectopic vas deferens opening into the ureter. AB - PURPOSE: A pediatric case of ectopic vas deferens opening into the ipsilateral ureter is reported. METHODS/RESULTS: Ectopic opening of the vas deferens was detected incidentally at ureterocystotneostomy being performed for vesicoureteral reflux to the solitary kidney of a 7-month-old boy with anorectal anomaly. CONCLUSIONS: The preservation of the patency of the seminal tract is recommended. PMID- 10375195 TI - Adenovirus-mediated direct gene therapy with bone morphogenetic protein-2 produces bone. AB - The need to improve bone healing permeates the discipline of orthopedic surgery. Bone morphogenetic proteins (BMPs) are capable of inducing ectopic and orthotopic bone formation. However, the ideal approach with which to deliver BMPs remains unknown. Gene therapy to deliver BMPs offers several theoretical advantages over implantation of a recombinant BMP protein, including persistent BMP delivery and eliminating the need for a foreign body carrier. A replication defective adenoviral vector was constructed to carry the rhBMP-2 gene (AdBMP-2). The direct in vivo gene therapy approach was applied in both immunodeficient and immunocompetent animals to produce intramuscular bone as early as 2 weeks following injection. Radiographic and histologic analysis revealed radiodense bone containing mature bone marrow elements. Adenovirus-mediated delivery of a marker gene (beta-galactosidase) into control animals produced no bone but indicated the cells transduced with the AdBMP-2 vector. Furthermore, comparisons between immunodeficient and immunocompetent animals illustrated the magnitude and significance of the immune response. Gene therapy to deliver BMP-2 has innumerable potential clinical applications from bone defect healing to joint replacement prosthesis stabilization. This study is the first to establish the feasibility of in vivo gene therapy to deliver active BMP-2 and produce bone. PMID- 10375196 TI - Interconversion potential of cloned human marrow adipocytes in vitro. AB - Information on the interconversion potential of adipocytes and other end cells characteristic of the stromal fibroblastic cell lineages, key in the understanding of bone turnover in metabolic diseases such as osteoporosis, is limited. The object of the present study was: i) to isolate relatively pure populations of adipocytes from human bone marrow; ii) to clone single adipocytes from these populations; and iii) to examine in vitro the interconversion potential of the progeny of these single-cloned adipocytes between the osteogenic and adipogenic phenotypes. Adipogenic colonies were isolated from the low-density floating fraction of normal bone marrow cells cultured in adipogenic media for 4 days. Single adipocytes were isolated and cloned by limiting dilution. Cloned adipocytes were found to dedifferentiate into fibroblast-like cells, and subsequently to differentiate into two morphologically distinct cell types: osteoblasts and adipocytes in appropriate culture systems. The adipocytic phenotype was confirmed by morphology, oil red O staining, and immunocytochemistry using antiserum to aP2. The osteogenic phenotype was confirmed by alkaline phosphatase, osteocalcin immunostaining using specific osteocalcin antiserum, and formation of mineralized cell aggregates. These findings demonstrate the extent of plasticity between the differentiation of adipocytic and osteogenic cells in human bone marrow stromal cell cultures. We have shown the ability of isolated clonal adipogenic cells to redifferentiate into cells of the osteogenic and adipogenic lineage and the interconversion potential of human marrow stromal cells in vitro. These results provide further evidence that the osteogenic and adipogenic cells share a common multipotential precursor. PMID- 10375197 TI - c-Myc and Mxi1 immunoreactivities in the calcifying areas of the epiphyseal-plate cartilage matrix of growing rats. AB - We looked for the protooncogene protein, c-Myc, its dimerization partner, Max, and the repressors of its transactivation activity, Mad1 and Mxi1, in the epiphyseal-plate cartilage matrix of growing rats by immunocytochemistry in the electron microscope. c-Myc and Mxi1 immunoreactivities were found in the calcifying areas of the cartilage matrix only. There was no immunolabeling in response to anti-Max or anti-Mad1 antibodies. Mxi1 immunoreactivity was mainly in the early calcifying areas, in the calcification front and ahead of it, whereas c Myc immunoreactivity was essentially in the incompletely calcified regions of the matrix. The two immunolabelings occurred mainly over the large type II collagen fibrils of the cartilage matrix and over the thin filaments connecting them. c Myc and Mxi1 immunoreactivities were rarely found along the dark cristallites. There was no immunolabeling associated with the matrix vesicles, or in their immediate surroundings. The data suggest that the protooncogene proteins, c-Myc and Mxi1, could be implicated in the calcification involving type II collagen fibrils of the epiphyseal-plate cartilage. The absence of Max immunoreactivity from the calcifying cartilage matrix raises the question of whether there are other c-Myc- and Mxi1-dimerization partners. PMID- 10375198 TI - Transforming growth factor beta (TGF-beta) levels in the conditioned media of human bone cells: relationship to donor age, bone volume, and concentration of TGF-beta in human bone matrix in vivo. AB - Transforming growth factor-beta (TGF-beta) is thought to play an important role in human bone remodeling. In the present study, we examined constitutive differences in TGF-beta levels in primary bone cell cultures from the iliac crest of 112 women, aged 28-79 years. TGF-beta1 was the major TGF-beta isoform in the conditioned media, as determined by neutralizing TGF-beta activity with specific antibodies against TGF-beta1-3 in the mink lung cell bioassay, and by enzyme linked immunoassay (ELISA). TGF-beta1 levels in the conditioned media did not change with donor age. There was a lack of association between TGF-beta levels in vitro and the concentration of matrix-associated TGF-beta in vivo. TGF-beta1 levels failed to be associated with the local trabecular bone volume in the complete study population (r = +0.15, p = 0.16, n = 89). A significant association between TGF-beta1 levels and bone volume was present in premenopausal women (r = +0.39, p = 0.02, n = 33), but was largely accounted for by the two samples with the highest TGF-beta concentrations. In conclusion, our data suggest that TGF-beta1 is the major TGF-beta isoform produced by human bone cells in vitro, and that the constitutive secretion of TGF-beta by bone cells does not change with age. Whether constitutive differences in TGF-beta secretion may be a determinant of human bone mass remains to be clarified in further studies. PMID- 10375199 TI - Differential regulation of syndecan expression by osteosarcoma cell lines in response to cytokines but not osteotropic hormones. AB - Bone cells are regulated by interactions with both growth factors and components of the extracellular matrix (ECM). Syndecans are cell-surface heparan sulfate proteoglycans known to play a role in cell adhesion and migration, and binding of growth factors. This study was performed to investigate the expression of syndecans by osteoblasts. Reverse transcription-linked polymerase chain reaction (RT-PCR) and Northern analysis detected syndecan transcripts in the human osteosarcoma cell lines MG-63, TE-85, SaOS-2, and U2OS; human osteoblast-like cells; rat calvarial osteoblasts; and in human bone. Western blot analysis of proteoglycans from MG-63 and TE-85 cells detected multiple heparan sulfate proteoglycan core proteins consistent with syndecan expression. Regulation of syndecan-1, -2, and -4 expression was investigated in TE-85, MG-63, and SaOS-2 cells, in response to interleukin (IL)-1beta, and IL-6, parathyroid hormone [PTH(1-34)], and 1,25(OH)2-vitamin D3. Northern analysis demonstrated that in the osteosarcoma cell lines there was no regulation of syndecan transcript levels in response to PTH(1-34) or 1,25(OH)2-vitamin D3 for 24 or 48 h. In contrast, when MG-63 and SaOS-2 cells were incubated with IL-1beta (0.01-10 ng/mL) and IL-6 (0.1 50 ng/mL) there was a dose-dependent decrease in mRNA levels for syndecan-1 and 2 at 24 and 48 h, but in response to IL-1beta upregulation in the levels of syndecan-4 transcripts. In addition, Northern analysis was performed on RNA isolated from neonatal rat calvarial osteoblasts cultured under conditions that promote osteogenesis for 0, 5, 13, 21, and 35 days. Syndecan-1 expression was observed to decrease during the culture period, syndecan-2 transcript levels increased, and there appeared to be no overall change in syndecan-4 levels. Controlled expression of syndecans by cells of the osteoblast lineage may be important in the regulation of osteoblastic proliferation and differentiation. PMID- 10375200 TI - Osteoconduction in large macroporous hydroxyapatite ceramic implants: evidence for a complementary integration and disintegration mechanism. AB - Large, cylindrical implants of a porous calcium phosphate ceramic ("hydroxyapatite" starting material, HAC) were used to replace far greater than critical-sized sections of the midshaft of sheep tibiae and retrieved at 2 and 9 months; external fixation was used in the first 5 months. Excellent clinical function of these implants was reported in a previous study. The material retrieved was embedded in PMMA, and blocks were sectioned and surfaces were polished and carbon coated prior to study using digital backscattered electron (BSE) imaging. Detailed scanning electron microscopy study of the pattern of osseointegration of the implanted material at early (2 months) and late (9 months) timepoints revealed a previously unrecognized pattern of integration/disintegration of this implant material in tandem with bone growth. We conclude that bone adaptation to the HAC leads to its fracture and that the newly generated surfaces are equally osteoconductive. This leads to a self propagating, self-annealing system in which defects in the HAC are mended by intercalation of bone. PMID- 10375201 TI - Trabecular remodeling processes in the ovariectomized rat: modified node-strut analysis. AB - The purpose of this study was to evaluate the trabecular bone remodeling processes in ovariectomized rats, focusing on diminishing trabecular connectivity. We used modified node-strut analysis defining three areas in the trabecular surfaces for the three-dimensional understanding of trabecular resorption derived from two-dimensional conventional sections, in addition to conventional bone histomorphometry and node-strut analysis. Seven-month-old female Wistar rats were used and treated with bilateral ovariectomy (ovx) and sham operation. Six rats in each group were examined at 4 and 8 weeks. We prepared undecalcified sections from the left tibiae with Villanueva bone and Goldner stains. We divided the trabecular bone surfaces (BS) into three areas: node (Nd), terminus (Tm), and strut (St), and measured the bone resorption and formation parameters, including eroded surface (ES), osteoclast surface (Oc.S), osteoid surface (OS), and double-labeled surface (dLS) in each defined area. In conventional bone histomorphometry, the ovx group showed high turnover osteopenia compared with the sham operation group. In node-strut analysis, the ovx group showed significantly lower values for node-related parameters than did the sham operation group. In the modified node-strut analysis, bone resorption parameters in the ovx group showed significantly higher values, particularly for strut and terminus-eroded surfaces (StES/BS, TmES/BS), and for each area of osteoclast surface (NdOc.S/BS, TmOc.S/BS, and StOc.S/BS) compared with the sham operation group. Bone formation parameters in the ovx group also showed significantly higher values, particularly for strut and terminus osteoid surfaces (TmOS/BS, StOS/BS), and for each area of double-labeled surface (NddLS/BS, TmdLS/BS, and StdLS/BS) compared with the sham operation group at 4 weeks. At 8 weeks, each area of bone formation parameter in the ovx group showed significantly higher values than that in the sham operation group. These results suggest that in the ovx group, the trabecular plates became perforated and the perforative cavities progressively enlarged, and/or the edges of plates were eroded regardless of elevated bone formation, resulting in diminished trabecular connectivity, and the node area might not be influenced relatively by bone remodeling in the early resorption. PMID- 10375202 TI - Comparison of three methods for estimation of bone resorption following ovariectomy in the distal femur and the proximal tibia of the rat. AB - The study of estrogen-deficient bone loss requires accurate assessment of bone resorption; however, there has been considerable variance in the values reported for this variable. We compared three techniques for estimating bone resorption in the adult rat at four anatomical sites, the metaphysis and epiphysis of the distal femur and the proximal tibia. The techniques include an osteoclast morphology-based method (VK-Oc), a bone-surface-based method (Pit-Oc), and an enzymatic method (AP-Oc). Thirty 6-month-old rats were either ovariectomized (ovx) or sham operated (sham) and killed at 0, 9, or 18 days postoperation. Each method was analyzed for variance in the ovary-intact groups and for the ability to detect the increase in osteoclast surface known to occur following ovariectomy. A 50-fold variation for the estimation of extent of osteoclast surface was entirely accounted for by these three methods for osteoclast estimation. The VK-Oc method was the most consistent for discriminating levels of osteoclast surface between ovary-intact and ovariectomized rats, detecting an increase at three of the four sites. There was no difference between the methods in their ability to produce consistent values for ovary-intact groups. The Pit-Oc method produced the largest numerical difference between the two operation groups with a 2.5-3-fold increase compared with 1.25-1.5-fold for the other methods (p < 0.001). However, the greater variance associated with this method limited the ability to detect the increase in osteoclast surface following ovariectomy. The AP-Oc method lacked the sensitivity of the other two methods. The oophorectomy induced increase of osteoclast surface in the metaphysis of distal femur was larger and more consistently demonstrated than increases at the other sites. PMID- 10375203 TI - A model to monitor the efficacy of alendronate treatment in women with osteoporosis using a biochemical marker of bone turnover. AB - Markers of bone turnover have been suggested to be useful in monitoring the long term efficacy of antiresorptive therapy on bone mineral density (BMD). In this study, we developed a new model based on the combination of a marker level and its percent change at 6 months of therapy to predict long-term response in BMD. Serum bone alkaline phosphatase (BAP) was measured in 307 late postmenopausal women (mean age 64 years) with osteoporosis enrolled in a 2 year placebo controlled trial of the bisphosphonate alendronate (10 mg/day). Under treatment, the maximal decrease was observed at 6 months (-44%) with no further change during the 2 year period. Both BAP levels at 6 months and percent BAP change at 6 months correlated with the percent change of spine BMD at 2 years (r = -0.54 and 0.53, respectively, p < 0.001 for both). Logistic regression analysis showed that BAP levels and percent BAP change at 6 months are independent predictors of long term positive BMD response, defined as > or = 3% increase in spine BMD at 2 years. The most relevant clinical option that could lead to therapeutic adjustment is likely to be an accurate identification of nonresponders, and thus predictive models need to be highly specific. For a 90% specificity, the combination of both the percent change and BAP levels at 6 months resulted in a significantly (p < 0.05) higher sensitivity (72%) than using percent BAP change (61%) or BAP level at 6 months (59%) alone. This combination model was also more effective than using the least-significant change (a decrease of BAP at 6 months of >44%) based on the within-patient variability in the placebo group. In the combination model, positive BMD responders vs. nonresponders could easily be distinguished by a line on a two-scale graph (BAP level at 6 month vs. percent BAP change at 6 months). In conclusion, the combination of BAP level and of its percent change after 6 months of treatment in a logistic model improved the prediction of the long-term BMD response to alendronate treatment compared with percent BAP change alone. This new model may be useful for quick and accurate identification of noncompliant patients (i.e., nonresponders) vs. responders to alendronate treatment, although prospective studies are required to determine accurately the rate of false positives and false negatives. Because this model is independent of the study design, it should be broadly applicable. PMID- 10375204 TI - Ultrasonic measurement of the calcaneus in Polish normal and osteoporotic women and men. AB - In this cross-sectional study, 964 subjects (740 postmenopausal women and 224 men) who underwent ultrasound measurement of the calcaneus at the outpatient osteoporotic clinic in the years 1993-1996 were evaluated. The aim of the study was to compare the influence of age, years since menopause (YSM), and body size on ultrasound variables in normal and osteoporotic male and female populations as well as to assess the ability of quantitative ultrasound (qUS) to discriminate between healthy and osteoporotic individuals. The subjects were divided into four age-matched groups: normal women (n = 601, mean age 56.3 +/- 4.8 years); osteoporotic women (n = 139, mean age 56.5 +/- 4.8 years); normal men (n = 148, mean age 56.2 +/- 10.2 years); and osteoporotic men (n = 76, mean age 56.5 +/- 10.7 years). Persons with a history of a disease and/or medications known to affect bone metabolism were excluded. Broadband ultrasound attenuation (BUA, in decibels per megahertz) and speed of sound (SOS, in meters per second) were measured using Achilles device (Lunar, Madison, WI). Short- and long-term in vitro coefficients of variation (CVs) were: 1.23% and 0.54% for BUA and 0.12% and 0.14% for SOS, respectively. Short-term in vivo CVs were: in women, BUA 1.8% and SOS 0.22%; and, in men, 2.48% and 0.33%, respectively. SOS and BUA values were significantly higher in healthy men (1517.5 +/- 35.3 m/sec, 114.0 +/- 13.3 dB/MHz) than in healthy women (1511.1 +/- 25.6 m/sec, 108.7 +/- 9.5 dB/MHz) (p < 0.000001). The two ultrasound variables had higher values in osteoporotic men (SOS = 1492.6 +/- 24.6 m/sec, BUA = 106.1 +/- 11.6 dB/MHz) in comparison to osteoporotic women (SOS = 1490.4 +/- 19.5 m/sec, BUA = 103.2 +/- 8.6 dB/MHz), but the differences did not achieve significance. In both genders, ultrasound parameters were significantly lower in osteoporotic groups (p < 0.000001). The following age-adjusted odds ratios were obtained: in women, 1.7 (95% CI 1.42 2.03) for BUA, and 2.3 (95% CI 1.87-2.81) for SOS; in men, 1.05 (95% CI 0.03 2.07) for BUA, and 2.13 (95% CI 0.77-3.49) for SOS. ROC analyses performed in both genders showed the following area under the curve data: in women, 0.66 for BUA, and 0.74 for SOS; and, in men, 0.66 for BUA, and 0.71 for SOS. Multiple regression analysis showed age to be the main negative determinant of both ultrasound variables in healthy women, and YSM in osteoporotic women. In both genders, weight was found to have a positive influence on SOS and BUA values, whereas the effect of height was weaker and generally negative. It can be concluded that ultrasound measurement at the calcaneus is able to discriminate between normal male/female and osteoporotic male/female populations in a similar manner. Women had greater rates of decrease in BUA and SOS with age compared with rates in men, and the decrease was greater in normal individuals in both genders. Distinct gender-related differences were noted in regard to correlations of ultrasound parameters with body size. PMID- 10375205 TI - Primary difficulties in quantitative backscattered electron (BSE) imaging. PMID- 10375206 TI - Victor Arthur Rice, 1890-1964: a brief biography. PMID- 10375207 TI - Sire marbling score expected progeny difference and weaning weight maternal expected progeny difference associations with age at first calving and calving interval in Angus beef cattle. AB - Field records from the American Angus Association were used to study the associations of sire marbling score EPD and sire weaning weight maternal (milk) EPD with age at first calving (AFC) and calving interval (CI). Cows were selected based on the accuracy of their sire's milk (> or =.7) or marbling (> or =.6) EPD. The data were screened using biological constraints, and regression models were used to identify records that were greater than 5 SD from the mean. The AFC was modeled for both milk and marbling data sets to account for effects of year, sire EPD, and their interaction. The CI was subdivided into first, second, and mature calving interval traits and modeled to account for state, year, calf sex, calf birth weight (BW), calf weaning weight (WW), sire EPD, and interactions of EPD with year and state. Derivative-free REML was used to estimate heritability and genetic correlations for AFC and CI. Sire milk EPD and marbling EPD were predictors of AFC (P < .001); however, pooled estimates were unreliable because of state x EPD interactions (P < .001). Increases in sire milk EPD resulted in reductions in AFC; however, there was no consistent pattern to effects of marbling EPD increases. Models accounted for < 8% of variation in AFC. Sire milk EPD was not a predictor of first, second, or mature CI (P > .1). Sire marbling score EPD was not a predictor of second, or mature CI (P > .1); however, it was associated (P = .059) with first CI, although regression estimates varied across states and prevented pooling. The BW, sex, and WW were predictors of CI (P < .001). Increases in BW resulted in longer mature CI, and mature CI decreased as WW increased. The AFC was heritable (.22), and CI traits had heritabilities ranging from .01 to .03. The AFC was genetically correlated with first CI (-.6) and mature CI (-.93). Genetic correlations between CI traits were uninterpretable because of low additive genetic variances. In conclusion, sire marbling score and milk EPD do not seem to be reliable predictors of AFC or CI. The BW and WW have significant but small effects on AFC and CI. Selection for AFC is possible, but earlier calving heifers may have longer calving intervals. PMID- 10375208 TI - Effects of continuous ivermectin treatment from birth to puberty on growth and reproduction in dairy heifers. AB - The effect of continuous ivermectin treatment from birth to puberty on growth and reproductive performance was studied in Holstein heifer calves grown on pastures in comparison to naturally nematode-infected, untreated animals. Ivermectin effectively abated the presence of nematode eggs in feces. Eggs per gram (EPG) in parasitized animals increased rapidly from wk 12 to 18 of age and then decreased. Animals treated with ivermectin grew faster than untreated ones, and differences in body weight became significant at 6 wk of life, even before eggs appeared in the feces of either treatment group. Ivermectin-treated heifers reached puberty 3 wk earlier than infected ones as assessed with serum progesterone concentrations (ivermectin, 30.4 +/- .8 vs untreated, 33.7 +/- 1.3 wk of age). This delay was not directly related to body weight. In addition, pelvic area at 39 wk and at 15 mo of age was increased in treated heifers (8 and 11%, respectively) compared with parasitized animals. No differences in the wither heights were observed. We conclude that ivermectin treatment in dairy heifers may increase growth rate during development, advance the onset of ovarian function, and positively affect yearling pelvic area. PMID- 10375209 TI - Repeatability of ultrasound-predicted percentage of intramuscular fat in feedlot cattle. AB - We used data from 144 bulls, heifers, and steers to determine the repeatability of ultrasound-predicted percentage of intramuscular fat and to study the effect of repeated measurements on the standard error of prediction. Animals were scanned at an average age of 433 d by a certified technician. Individual bulls, heifers, and steers were scanned five to six times each with two Aloka 500-V machines, and the percentage of intramuscular fat was predicted from two regions of interest within an image. Variance components and repeatability values were computed for the overall data and by machine, region of interest, and sex. Animals were broadly divided into two groups based on mean ultrasound-predicted percentage of intramuscular fat. Variance components and repeatability values were then estimated within each group. The overall repeatability of ultrasound predicted percentage of intramuscular fat was .63 +/- .03. Differences in the repeatability values between machines and between regions of interest were not different from zero (P > .05). Bulls showed a lower within-animal SD of .82% as compared to .97 and 1.02% for steers and heifers, respectively. However, steer ultrasound-predicted percentage of intramuscular fat measures were more repeatable (P < .05) than those of bulls and heifers. The difference in repeatability between bull and heifer measures was not important (P > .05). Animals with mean ultrasound-predicted percentage of intramuscular fat less than 4.79% showed less repeatable measures (P < .05) than those with means above 4.79%. The image variance contributed to nearly 70% of the total variance of observations within an animal. Standard error of animal mean measures showed a 50% reduction when the number of images per animal increased to four. Therefore, we concluded that increasing the number of images per animal plays a more significant role in reducing the standard error of prediction than taking multiple measurements within a single image. PMID- 10375210 TI - Effects of individual housing design and size on behavior and stress indicators of special-fed Holstein veal calves. AB - The objectives of this study were to determine effects of housing design (calves tethered in open stalls vs untethered in individual pens) and widths of 56, 66, and 76 cm (2 x 3 factorial arrangement of treatments) on indicators of stress and behavior in special-fed veal calves. Three production cycles (groups) were used, each with 36 Holstein bull calves. Calves (n = 108) were randomly allotted to treatments upon arrival at the facility. Blood samples were collected four times (wk 4, 9, 13, and 18) during the 18-wk production cycle. Blood serum values for cortisol and alpha1-acid glycoprotein (AGP) exhibited few treatment differences. Blood leukocyte differential counts at 4 and 18 wk (segmented neutrophils [N], banded neutrophils, lymphocytes [L], basophils, and the N:L ratio) were not different (P > .05) among housing designs or widths. However, there were differences (P < .05) in monocytes and eosinophils during the 28-d period after arrival; calves in stalls 76 cm wide had the greatest percentage of both leukocytes, and calves in the 66-cm stalls had the lowest monocyte percentage. Calves were recorded on videotape during wk 4, 13.5, and 18 to determine frequencies and durations of postures and behaviors (e.g., lying, standing, chewing, tongue playing, grooming, and investigative activities). There were no consistent differences (P > .05) in postures or behaviors among calves in different housing designs or widths. Calves spent approximately 71 and 31% in lying and standing positions, with no preference for the right or left side while recumbent. There was a tendency for calves in wider stalls or pens at wk 9 and 18 to exhibit more self-grooming activities. Tongue playing and investigative and chewing activities were exhibited in all treatments, but no differences (P > .05) were observed. However, calves housed in the 56-cm pens displayed difficulty in changing from lying to a standing position and were unable to extend one or more legs while recumbent. Even though there were few differences in behavioral, physiological, growth, or anatomical traits in this study, further increases in age and(or) weight of finished calves will require a reassessment of the appropriateness of individual veal calf housing design and dimensions. PMID- 10375211 TI - Influence of norgestomet in combination with gonadotropins on induction of estrus and ovulation in prepubertal gilts. AB - Our objective was to determine whether priming with the progestogen norgestomet for 9 d would enhance estrual and ovulatory responses of prepubertal gilts to PG600 (400 IU eCG + 200 IU hCG). Gilts (140 to 190 d old) were assigned by litter, age, and weight to one of three treatments: 1) 9 d of norgestomet implant with an injection of PG600 after implant removal on d 9 (N+PG; n = 43); 2) no implant and an injection of PG600 on d 9 (PG; n = 36); or 3) neither implant nor PG600 (control; n = 29). Beginning on d 0, gilts were exposed once daily to a boar and checked until estrus was observed or until d 45 after the start of the experiment. Ovaries were examined for number of corpora lutea (CL) after estrus or at 45 d. Greater proportions of N+PG (63%, P < .05) and PG (69%, P < .01) gilts expressed estrus than did controls (34%), but proportions did not differ between N+PG and PG (P > .10). Among gilts in estrus following treatment with N+PG or PG, 100% showed estrus within 6 d after PG600 injection. For gilts that expressed estrus within 45 d, the average age at estrus was reduced (P < .05) by PG to 172 +/- 2 d compared with 182 +/- 4 d for controls. Average age at estrus did not differ (P > . 10) between PG and N+PG (177 +/- 2 d). Greater proportions of N+PG (82%; P < .001) and PG (65%; P < .001) gilts ovulated than controls (13%), but proportions did not differ between N+PG and PG (P > .10). The number of CL (20 +/- 2) was not affected by treatment and ranged from 2 to 71. There was no increase in ovarian cysts in response to treatment. Results indicated that norgestomet before PG600 did not enhance estrus expression or ovulation compared with PG600 alone, but use of PG600 increased the proportions of gilts that expressed estrus and ovulated compared with controls. PMID- 10375212 TI - Biologically based coefficients for partitioning lamb and wool production costs. AB - Global competition for selling lamb and wool requires sheep producers to effectively prioritize wool and lamb production. Both products are produced simultaneously, and this makes it difficult to differentially assess costs and net returns. This study addressed this issue by developing coefficients for use in financial analysis of lamb and wool profitability across and within five regions of the United States (Eastern, Midwestern, Intermountain West, Great Basin, and Texas). A procedure was developed using a sheep simulation model to partition the amount of nutrients used for lamb and wool production and then using the partitioning information to determine the proportion of costs to be assessed to lamb and wool production. Blackface breed types, when managed without nutritional limitations, had lamb and wool coefficients of .90 and .10, respectively. Wool breeds with unlimited and limited nutrition had lamb coefficients between .80 and .20 and .60 and .40, respectively. In-depth analysis of the Great Basin system indicated that wool and its improvement may contribute as much, if not more, under certain pricing conditions, as increased prolificacy to enterprise profitability. With low lamb prices ($1.32/kg) and moderate wool prices, the partial net returns for a Merino x Rambouillet were higher than the tested prolific breed type ($13.85 vs $11.27/ewe, respectively). This ranking was reversed under high ($2.31/kg) lamb prices ($47.90 vs $55.22, respectively). The derived method provides a basis for analyzing, comparing, and making management and breeding objective decisions. PMID- 10375213 TI - A region on bovine chromosome 15 influences beef longissimus tenderness in steers. AB - A genome scan was conducted using 196 microsatellite DNA markers spanning 29 autosomal bovine chromosomes and Warner-Bratzler shear force collected at d 2 and 14 postmortem on steaks from the longissimus muscle of 294 progeny from one Brahman x Hereford bull mated to Bos taurus cows to identify QTL for beef tenderness. One QTL was identified and located 28 cM (95% confidence interval is 17 to 40 cM) from the most centromeric marker on BTA15. The QTL interacted significantly with slaughter group. The difference in shear force of steaks aged 14 d postmortem between progeny with the Brahman paternally inherited allele vs those with Hereford was 1.19 phenotypic standard deviations (explained 26% of phenotypic variance) for one slaughter group and was not significant for three other slaughter groups. Apparently, unknown environmental factors present for three of the four slaughter groups were capable of masking the effect of this QTL. The sensitivity of the QTL effect to environmental factors may complicate utilization of markers for genetic improvement. Future research to elucidate the cause of the QTL x slaughter group interaction may lead to improved strategies for controlling variation in meat tenderness via marker-assisted selection, postmortem processing, or live animal management. PMID- 10375214 TI - Effect of missing data on the estimate of average daily feed intake of growing pigs. AB - The effect of missing records for feed intake per day (FID) on the estimate of average daily feed intake (DFI) during the test period of individual pigs was studied. Data from 192 growing pigs tested with single-space computerized feeding stations during an average of 93 d were used. True DFI was computed by averaging FID records per pig, individually. A first- and third-degree polynomial and a nonlinear function were fit to FID records per pig to estimate DFI by averaging estimated FID records per pig, individually. The three functions showed small differences for goodness of fit. Missing FID records were simulated by random as well as period-wise deletions of FID records. The effect of missing FID records was judged on the Pearson correlation between true and estimated DFI. Deleting randomly up to 70% of FID records per pig before fitting each function reduced this correlation only from 1.00 to .96 for each function. Deleting 25 successive FID records (approximately 27% of records) before fitting reduced the correlation to values ranging from .92 to .96 and from .59 to .96 for the first- and third order degree polynomial function, respectively, and from .80 to .97 for the nonlinear function. Using iteratively reweighted least squares regression methods to exclude undesirable effects of outlier values gave similar results for the effect of missing FID records on estimated DFI. Results imply that considering incorrect FID records as missing is a good alternative for adjusting incorrect data in combination with using functions to estimate DFI of growing pigs. Use of a first-degree polynomial function is recommended. Moreover, use of functions enables a more efficient use of feeding stations by recording feed intake data during only parts of the test period. PMID- 10375215 TI - A primary screen of the bovine genome for quantitative trait loci affecting carcass and growth traits. AB - A primary genomic screen for quantitative trait loci (QTL) affecting carcass and growth traits was performed by genotyping 238 microsatellite markers on 185 out of 300 total progeny from a Bos indicus x Bos taurus sire mated to Bos taurus cows. The following traits were analyzed for QTL effects: birth weight (BWT), weaning weight (WW), yearling weight (YW), hot carcass weight (HCW), dressing percentage (DP), fat thickness (FT), marbling score (MAR), longissimus muscle area (LMA), rib bone (RibB), rib fat (RibF), and rib muscle (RibM), and the predicted whole carcass traits, retail product yield (RPYD), fat trim yield (FATYD), bone yield (BOYD), retail product weight (RPWT), fat weight (FATWT), and bone weight (BOWT). Data were analyzed by generating an F-statistic profile computed at 1-cM intervals for each chromosome by the regression of phenotype on the conditional probability of receiving the Brahman allele from the sire. There was compelling evidence for a QTL allele of Brahman origin affecting an increase in RibB and a decrease in DP on chromosome 5 (BTA5). Putative QTL at or just below the threshold for genome-wide significance were as follows: an increase in RPYD and component traits on BTA2 and BTA13, an increase in LMA on BTA14, and an increase in BWT on BTA1. Results provided represent a portion of our efforts to identify and characterize QTL affecting carcass and growth traits. PMID- 10375216 TI - Identification of quantitative trait loci affecting female reproductive traits in a multigeneration Meishan-White composite swine population. AB - A multigeneration crossbred Meishan-White composite resource population was used to identify quantitative trait loci (QTL) for age at first estrus (AP) and the components of litter size: ovulation rate (OR; number of ova released in an estrous period) and uterine capacity (UC). The population was established by reciprocally mating Meishan (ME) and White composite (WC) pigs. Resultant F1 females were mated to either ME or WC boars to produce backcross progeny (BC) of either 3/4 WC 1/4 ME or 1/4 WC 3/4 ME. To produce the next generation (F3), 3/4 WC 1/4 ME animals were mated to 1/4 WC 3/4 ME animals yielding half-blood (1/2 WC 1/2 ME) progeny. A final generation (F4) was produced by inter se mating F3 animals. Measurements for AP and OR were recorded on 101 BC, 389 F3, and 110 F4 gilts, and UC data were from 101 BC and 110 F4 first parity litters. A genomic scan was conducted with markers (n = 157) spaced approximately 20 cM apart. All parental, F1, BC, and F4 animals but only 84 F3 animals were genotyped and included in this study. The QTL analysis fitted a QTL at 1-cM intervals throughout the genome, and QTL effects were tested using approximate genome-wide significance levels. For OR, a significant (E[false positive] < .05) QTL was detected on chromosome 8, suggestive (E[false positive] < 1.0) QTL were detected on chromosomes 3 and 10, and two additional regions were detected that may possess a QTL (E[false positive] < 2.0) on chromosomes 9 and 15. Two regions possessed suggestive evidence for QTL affecting AP on chromosomes 1 and 10, and one suggestive region on chromosome 8 was identified for UC. Further analyses of other populations of swine are necessary to determine the extent of allelic variation at the identified QTL. PMID- 10375217 TI - Direct measurements of methane emissions from grazing and feedlot cattle. AB - Methane (CH4) emissions from animals represent a significant contribution to anthropogenically produced radiatively active trace gases. Global and national CH4 budgets currently use predictive models based on emission data from laboratory experiments to estimate the magnitude of the animal source. This paper presents a method for measuring CH4 from animals under undisturbed field conditions and examines the performance of common models used to simulate field conditions. A micrometeorological mass difference technique was developed to measure CH4 production by cattle in pasture and feedlot conditions. Measurements were made continuously under field conditions, semiautomatically for several days, and the technique was virtually nonintrusive. The method permits a relatively large number of cattle to be sampled. Limitations include light winds (less than approximately 2 m/s), rapid wind direction changes, and high-precision CH4 gas concentration measurement. Methane production showed a marked periodicity, with greater emissions during periods of rumination as opposed to grazing. When the cattle were grazed on pasture, they produced .23 kg CH4 x animal(-1) x d(-1), which corresponded to the conversion of 7.7 to 8.4% of gross energy into CH4. When the same cattle were fed a highly digestible, high-grain diet, they produced .07 kg CH4 x animal(-1) x d(-1), corresponding to a conversion of only 1.9 to 2.2% of the feed energy to CH4. These measurements clearly document higher CH4 production (about four times) for cattle receiving low-quality, high-fiber diets than for cattle fed high-grain diets. The mass difference method provides a useful tool for "undisturbed" measurements on the influence of feedstuffs and nutritional management practices on CH4 production from animals and for developing improved management practice for enhanced environmental quality. PMID- 10375218 TI - Behavior of steers grazing monocultures and binary mixtures of alfalfa and tall fescue. AB - Spectral analysis was used to relate dietary quality and herbage species to the behavior of grazing steers. Four .3-ha paddocks were established with either 'AU Triumph' tall fescue (F; Festuca arundinacea Schreb.), 'Apollo' alfalfa (A; Medicago sativa L.), 1/3 fescue and 2/3 alfalfa (2/3A), or 2/3 fescue and 1/3 alfalfa (1/3A). Each paddock was stocked with 10 to 16 steers and defoliated in 5 d. Three steers on each paddock carried vibracorders to monitor grazing time. Daily forage samples were taken in 10-cm layers and weighed. Esophageal extrusa were collected from fistulated steers to measure diet quality. Daily grazing time did not differ (P = .37) among treatments; however, steers grazing mixtures grazed numerically longer (1.4 h/d) than steers on monocultures. Spectral analysis revealed that steers grazing A and 2/3A had many daily meals of short duration, but steers grazing 1/3A and F consumed three meals daily at 8-h intervals. Throughout the 4.67-d grazing period, quality of the diet linearly declined in crude protein and herbage digestibility, linearly increased in neutral detergent fiber and cellulose, and exhibited quadratic changes in lignin and ash. For most quality values, the tall fescue monoculture differed from the others (P < .05). Steers selected diets with similar quality for the A, 2/3A, and 1/3A treatments. This study illustrates how differences in forage diets alter grazing behavior of steers. PMID- 10375219 TI - Effects of anaerobic digestion and additives to effluent or cattle feed on odor and odorant concentrations. AB - Odor intensity (5,437 observations), determined by human panelists (100 different panelists over the course of the experiment), and a number of chemical odorant concentrations were determined for manure-related samples (326) obtained from effluents from conventional stirred-tank reactor (CSTR) and fixed-film anaerobic digesters, effluents to which commercial additives or KMnO4 or H2O2 were added, and feces, urine, and mixed manure from cows fed a control or additive-containing diet. Mostly, samples were held in stoppered, Erlenmeyer flasks for 3 d at room temperature before evaluation by panelists and with chemical analyses, but shorter holding times also were tested. Anaerobic digestion reduced odor intensity linearly with increasing hydraulic retention time (HRT) up to 20 d; fixed-film digestion with 1.5- or 2.3-d HRT reduced odor intensity similarly to that observed with 10-d HRT in CSTR. Addition of commercial products and chemicals altered some odorant concentrations (e.g., ammonia) but did not reduce odor intensity; some products increased odor intensity. Addition of a commercial yeast-based product to a dairy cow diet had no detectable effect. The cow diet study showed that fresh urine and feces alone were less odorous than a mixed combination (manure). Fresh manure was less odorous than manure held for 3 d. Total phenol was the odorant most highly correlated with odor intensity. Individual and total volatile fatty acids also contributed. Ammonia did not seem to be a major contributor to odor in this data set. PMID- 10375220 TI - The effects of long-term administration of recombinant bovine somatotropin (Posilac) and Synovex on performance, plasma hormone and amino acid concentration, and muscle and subcutaneous fat fatty acid composition in Holstein Friesian bull calves. AB - The effect of recombinant somatotropin (rbST), Synovex (Syn), and their combination (rbST+Syn) on intact male calves was examined in an experiment that lasted an average of 238 d. Holstein-Friesian bull calves were allotted to one of four subtreatments (n = 14/treatment) in a factorial arrangement. There were two levels of rbST (0; rbST) and two levels of the estrogenic growth promoter Synovex (0; Syn). The rbST was administered once every 2 wk as injections of 500 mg of Posilac. Synovex (C and S) was implanted at 90-d intervals. The animals were fed for ad libitum consumption a diet with a metabolizable energy concentration of 11.7 MJ/kg DM and 15% crude protein. The hot carcasses were weighed after the removal of kidney, pelvic, and cod fats, which were weighed separately. The 12th rib cut was saved for analysis. Average daily gain and feed conversion efficiency were increased by rbST treatment by 9% (P < .005) and 10% (P < .016), respectively. There was no significant effect of Syn treatment, nor was there a rbST x Syn interaction. The proportion of the fat of the large depots in the carcass was reduced by 34% (P < .0001) and in the longissimus muscle by 32% (P < .16) owing to the rbST treatment. The plasma concentrations of GH, insulin, and thyroxin were increased by rbST treatment (P < .001, P < .01, and P < .03, respectively). The concentration of IGF-I was not affected. Synovex had no effect on plasma hormone concentration. Plasma essential and nonessential amino acid concentrations were reduced by 14 and 9%, respectively, when rbST was injected. Concentrations of cholesterol and fatty acids in muscle and subcutaneous fat were not affected (P > .072) by the rbST treatment. Synovex increased the monounsaturated fatty acids (MUFA), and the combinaton of Syn with rbST reduced polyunsaturated fatty acid (PUFA) concentration in the longissimus muscle (at the 12th rib). The reduced muscle fat content of the rbST-treated animals was associated with a trend toward an increase in polyunsaturated fatty acids. PMID- 10375221 TI - Expression and location of IGF binding proteins-2, -4, and -5 in developing fetal tissues. AB - Insulin-like growth factors are associated with myogenesis in vivo, and their actions are mediated by IGF binding proteins (IGFBP). Sites of IGFBP production and their location during early development are not clear. The objective of this research was to examine the developmental expression and location of IGFBP-2, -4, and -5 mRNA and peptides in developing porcine skeletal muscle and liver. Pregnant pigs were euthanatized at various times postconception (pc). Developmental expression of IGFBP was evaluated using total RNA extracted from skeletal muscle and liver of 30-, 44-, 59-, 68-, 75-, 89-, and 109-d pc fetuses and from adult and neonatal pigs. Localization of IGFBP-2, -4, and -5 mRNA and peptides was examined by in situ hybridization and immunocytochemistry of muscle samples from contralateral pelvic limbs of each pig. Overall muscle IGFBP gene expression decreased (P < .05) with increasing age. Moreover, expression of liver IGFBP-2 and -5, but not of IGFBP-4, was greater (P < .05) during prenatal than during postnatal periods. The majority of immunoreactive IGFBP was located in developing muscle cells, with little localized to connective tissue, except at later stages of development. These data show that IGFBP-2, -4, and -5 expression is time- and tissue-dependent in fetal liver and muscle. PMID- 10375222 TI - Analysis of body composition changes of swine during growth and development. AB - This study was conducted to model the growth of carcass, viscera, and empty body components and component composition of pigs. Quantitative tissue and chemical composition of 319 swine, representative of barrows and gilts from five commercial genetic populations, was determined at eight stages of growth between 25 and 152 kg. After whole body grinding and carcass dissection, proximate analyses were performed to calculate concentrations of protein, lipid, moisture, and ash of carcass, viscera, empty body, carcass lean, and carcass fat. Linear and nonlinear equations were developed to investigate the growth patterns of each component. Nonlinear growth functions accounted for the greatest amount of variation in empty body protein, lipid, moisture, and ash mass. Differences (P < .05) existed between barrows and gilts for nearly all components investigated. Carcass lean and fat tissues significantly increased in lipid percentage and decreased in moisture percentage as live weight increased. There were significant changes in the ratio and composition of the tissues of barrows and gilts during growth. Nonlinear models fitted the data better than allometric equations for nearly all of the components investigated. PMID- 10375223 TI - Immunoblot analysis of calpastatin degradation: evidence for cleavage by calpain in postmortem muscle. AB - A negative correlation exists between calpastatin activity and meat tenderness. Therefore, it is important to determine the mechanism of calpastatin inactivation in postmortem skeletal muscle. Western immunoblot analysis was performed to determine the protease(s) responsible for degradation of muscle calpastatin during postmortem storage. To accomplish this, purified calpastatin was digested with different proteases in vitro, and their pattern of calpastatin degradation was compared with that of calpastatin degradation in postmortem muscle. Polyclonal antibodies raised in mice against recombinant bovine skeletal muscle calpastatin were used to monitor calpastatin degradation. Lamb longissimus was stored at 4 degrees C and sampled at 0, 6, 12, 24, 72, 168, and 336 h postmortem. Postmortem storage produced a discrete pattern of calpastatin degradation products that included immunoreactive bands at approximately 100, 80, 65, 54, 32, and 29 kDa. Undegraded calpastatin (130 kDa) was barely detectable after 72 h of postmortem storage at 4 degrees C, and no immunoreactive calpastatin was observed by 336 h postmortem. For in vitro proteolysis, lamb longissimus calpastatin (0 h postmortem) was purified using Affi-Gel Blue chromatography. Calpastatin was digested with m-calpain, mu-calpain, cathepsin B, proteasome, trypsin, or chymotrypsin. Each of these enzymes degraded calpastatin. Immunoreactive fragments resulting from digestion of calpastatin with m- and mu-calpain were similar to each other and closely resembled those observed during postmortem aging of lamb longissimus at 4 degrees C. Digestion of calpastatin with mu calpain reduced calpastatin activity. Degradation of calpastatin by other proteases resulted in unique patterns of immunoreactive fragments, distinct from that observed in longissimus. Thus, m- and(or) mu-calpain seem to be responsible for calpastatin degradation during postmortem storage of meat. PMID- 10375224 TI - Tenderness classification of beef: II. Design and analysis of a system to measure beef longissimus shear force under commercial processing conditions. AB - The objectives of this study were to evaluate the efficacy of a system for classifying beef for tenderness based on a rapid, simple method of measuring cooked longissimus shear force. Longissimus steaks (2.54 cm thick) were trimmed free of s.c. fat and bone and rapidly cooked using a belt grill. A 1-cm-thick, 5 cm-long slice was removed from the cooked longissimus parallel with the muscle fibers for measurement of shear force. Slices were sheared with a flat, blunt-end blade using an electronic testing machine. The entire process was completed in less than 10 min. Therefore, in commercial application, this process could be completed during the 10- to 15-min period that carcasses are normally held to allow the ribeye to bloom for quality grading. In Exp. 1, the repeatability of slice shear force (SSF), as determined by evaluation of duplicate samples from 204 A-maturity carcasses, was .89. In Exp. 2, A-maturity carcasses (n = 483) were classified into three groups based on SSF (< 23, 23 to 40, and > 40 kg) at 3 d postmortem that differed (P < .001) in mean trained sensory panel tenderness ratings (7.3 +/- .04, 6.4 +/- .06, and 4.4 +/- .20) and the percentages (100, 91, and 28%) of samples rated "Slightly Tender" or higher at 14 d postmortem. Therefore, this tenderness classification system could be used to accurately segregate beef carcasses into expected tenderness groups. Further research is needed to test the feasibility and accuracy of this system under a variety of commercial processing conditions. PMID- 10375225 TI - Influence of the three RN genotypes on chemical composition, enzyme activities, and myofiber characteristics of porcine skeletal muscle. AB - An experiment was conducted to evaluate the effects of the RN genotype on skeletal muscle characteristics in pigs sharing otherwise the same polygenic background. Animals were genotyped for RN on the basis of RTN (Rendement Technologique Napole) records using segregation analysis methods. Samples of longissimus (L) and semispinalis capitis (S) muscles were taken from 39 rn+/rn+, 38 RN-/rn+ and 37 RN-/RN- pigs slaughtered at 108 +/- 8.6 kg live weight. Activities of lactate dehydrogenase (LDH), citrate synthase (CS), and beta hydroxy-acyl-coenzyme A dehydrogenase (HAD) were measured on both muscles to assess glycolytic, oxidative, and lipid beta-oxidation capacities, respectively. Histological examinations and chemical analyses were performed on L muscle. The energetic metabolism of the white L muscle was more oxidative in RN-/RN- than in rn+/rn+ pigs, as shown by increased CS and HAD activities (P < .001), decreased LDH activity (P < .001), larger cross-sectional area of IIA (P < .05) and IIB-red (P < .05) fibers, higher relative area of IIA fibers ( P < .05), and lower relative area of IIB-white fibers (P < .001). No significant difference was found between heterozygous and homozygous carriers of the RN- allele, except for CS activity, which was lower in RN-/rn+ than in RN-/RN- pigs. In L muscle, the RN- allele led to a large increase in glycolytic potential (+3.5 phenotypic SD between homozygotes) and lightness (+.7 SD), and a decrease in ultimate pH, dry matter, and protein contents (-1.7 to -2 phenotypic SD for these three traits), with an almost completely dominant effect. No differences were found between genotypes for intramuscular fat and hydroxyproline contents. In the red S muscle, the presence of RN- had no influence on enzyme activities. These results indicate that the RN genotype greatly influences compositional and histochemical traits and metabolic enzyme activities in a muscle type-dependent manner, with a completely or incompletely dominant effect of the RN- allele. PMID- 10375226 TI - Postmortem proteolysis and calpain/calpastatin activity in callipyge and normal lamb biceps femoris during extended postmortem storage. AB - The present experiment was conducted to determine whether calpastatin inhibits only the rate, or both the rate and extent, of calpain-induced postmortem proteolysis. Biceps femoris from normal (n = 6) and callipyge (n = 6) lamb was stored for 56 d at 4 degrees C. Calpastatin activity was higher (P < .05) in the callipyge muscle at 0 and 14 d postmortem, but not at 56 d postmortem. The activity of mu-calpain did not differ between normal and callipyge biceps femoris at 0 and 56 d postmortem (P > .05), but was higher at 14 d postmortem in the callipyge muscle (P < 0.05). The activity of m-calpain was higher in the callipyge muscle (P < 0.05). Western blot analyses of titin, nebulin, dystrophin, myosin heavy chain, vinculin, alpha-actinin, desmin, and troponin-T indicated that postmortem proteolysis was less extensive in callipyge than in normal biceps femoris at all postmortem times. The results of this experiment indicate that calpastatin inhibits both the rate and extent of postmortem proteolysis. PMID- 10375227 TI - The energy content of barley fed to growing pigs: characterizing the nature of its variability and developing prediction equations for its estimation. AB - Currently, the pork industry attempts to formulate energy levels in swine diets to within a tolerance of 1.5%. This is difficult to achieve in practice when the energy content of primary ingredients fluctuates by up to 15%. This experiment was carried out to define the sources of variation in the energy content of barley and to develop a practical method to accurately estimate the DE and ME content of individual barley samples. Four samples of each of five covered barley varieties (AC Lacombe, B-1602, Bedford, Harrington, and Manley) were collected to obtain a range of quality within each variety. Five measurements were collected on each barley sample using 60 crossbred barrows in an apparent total tract digestibility study. The barrows, average BW of 35.3 kg, were housed in individual metabolism crates to facilitate separate collection of urine and feces. Five-day collection periods followed 5-d diet acclimation periods. Levels of total beta-glucan, ADF, CP, and starch (90% DM) in the 20 barley samples ranged from 2.7 to 4.5%, 4.5 to 9.2%, 10.8 to 15.1%, and 42.3 to 53.4%, respectively. The mean DE and ME content of the 20 samples were 2,934 and 2,857 kcal/kg (90% DM), respectively, and varied among samples by 15.2% (447 kcal). The complex structural cell wall carbohydrates seemed to have the greatest influence on the energy content of individual barley samples. The ADF fraction alone accounted for 85% of the total variation in energy content of the 20 samples. Converted into a prediction equation, DE = 3,526 - 92.8 x ADF (90% DM), the ADF content was used to estimate the DE content of barley with 85% accuracy. This experiment confirms the large variation in the energy content of barley, describes the factors that influence this variation, and presents equations based on chemical and(or) physical measurements that may be used to predict the DE and ME content of individual barley samples. PMID- 10375228 TI - Impact of amino acid nutrition during lactation on body nutrient mobilization and milk nutrient output in primiparous sows. AB - The impact of amino acid nutrition during lactation on body nutrient mobilization and milk nutrient output in primiparous sows was evaluated. Thirty-six sows, nursing litters of 13 pigs, were allocated daily 6 kg of a fortified corn-soybean meal diet containing a high (HP, 1.20% lysine) or low (LP, .34% lysine) protein content during a 23-d lactation. Dietary lysine concentration was achieved by altering the ratio of corn and soybean meal in the diet. The LP sows consumed less daily ME (14.2 vs 16.1 Mcal; P < .11) and daily lysine (16 vs 59 g; P < .01) than the HP sows. Daily litter weight gain was less (P < .01) for sows fed the LP vs HP diet, and the differences increased (P < . 01) as lactation progressed. The lower litter weight gain for the LP sows was reflective of the lower (P < .01) estimated milk DM, CP, and GE output of these sows. The LP sows lost more body weight (1.23 vs .21 kg/d; P < .01) during the initial 20 d of lactation. In the LP sows, 59% of the weight loss was protein, water, and ash, and 37% was fat. Weight loss in the HP sows was entirely accounted for by body fat mobilization, because these sows accrued body protein, water, and ash. Muscle myofibrillar breakdown rate was higher in LP sows than in HP sows (4.05 vs 2.80%/d; P < .01). On the basis of these data, dietary amino acid restriction during lactation increases maternal mobilization of proteinaceous tissue and reduces milk nutrient output. Maternal protein mobilization is maintained over the entire lactation even though milk output is decreased as lactation progresses. PMID- 10375230 TI - Failure of vitamin A to increase litter size in sows receiving injections at various stages of gestation. AB - To determine the effectiveness of a single injection of vitamin A to increase litter size, 1,375 sows were assigned randomly to 11 treatment groups (125 sows per treatment). Treatments included injection of 1 x 10(6) IU of vitamin A dissolved in corn oil at weaning or on d 0, 2, 6, 10, 13, 19, 30, 70, or 110 after breeding. Sows in the control group were injected with corn oil on corresponding days. A total of 396 sows were removed from the study following treatment or treatment assignment. Therefore, farrowing data were collected for 979 sows. Injection of vitamin A did not influence (P > . 10) total litter size, live litter size, litter weight, pig weight, number of runts, or number of mummies. Mean live litter size was 10.1 +/- .1 for all sows that farrowed in the experiment. Parity group affected total litter size, live litter size, live litter weight, and number stillborn (P < .01) but not pig weight, number of runts, number of mummies, or gestation length (P > . 10). In this study, a single injection of vitamin A at any time from weaning to farrowing did not increase litter size in sows. PMID- 10375229 TI - Impact of amino acid nutrition during lactation on luteinizing hormone secretion and return to estrus in primiparous sows. AB - The impact of the dietary amino acid regimen of primiparous sows on LH secretion and weaning-to-estrus interval was evaluated. Thirty-six sows, nursing litters of 13 pigs, were allocated daily 6 kg of a corn-soybean meal diet containing a high (HP, 1.20% lysine) or low (LP, .34% lysine) protein content during a 23-d lactation. Dietary lysine concentration was achieved by altering the ratio of corn and soybean meal in the diet. Plasma LH, ACTH, and estradiol-17beta were evaluated at 15 min, hourly, and at 6-h intervals, respectively, during 6-h periods on d 0, 5, 10, 15, and 20 of lactation. Sows were checked daily for estrus from weaning to 45 d postweaning. Sows fed the LP and HP diets consumed 4.41 and 4.98 kg of feed daily during lactation. The LP sows weighed less (P < .05), had lighter (P < .05) litters at weaning, and had (P < .05) extended weaning-to-estrus intervals. Mean and baseline LH concentrations and LH pulses/6 h were lower (P < .01) in LP sows, and the differences between LP and HP sows were established by d 10 of lactation. Plasma estradiol and ACTH concentrations were not altered by diet. Mean LH concentrations on d 5 and 10 of lactation were correlated (r = -.54 and -.56, respectively, P < .01) with weaning-to-estrus interval. Also, mean LH concentrations on d 10 were correlated (P < .05) with the magnitude of dietary lysine deficiencies relative to demand for milk synthesis on d 0 to 5 and d 5 to 10 (r = -.39 and -.49, respectively). Inadequate dietary amino acid intake in sows during early lactation results in lower LH secretion by d 10 postpartum and is associated with increased weaning-to-estrus interval. PMID- 10375231 TI - Nutritionally induced anovulation in beef heifers: ovarian and endocrine function preceding cessation of ovulation. AB - Angus x Hereford heifers were used to determine endocrine and ovarian function preceding nutritionally induced anovulation. Six heifers were fed to maintain body condition score (M), and 12 heifers were fed a restricted diet (R) until they became anovulatory. Starting on d 13 of an estrous cycle, heifers were given PGF2alpha every 16 d thereafter to synchronize and maintain 16 d estrous cycles. Ovarian structures of M and R heifers were monitored by ultrasonography daily from d 8 to ovulation (d 1 of the subsequent cycle) until R heifers became anovulatory. Concentrations of LH and FSH were quantified in serum samples collected every 10 min for 8 h on d 2 and 15 (48 h after PGF2alpha), and estradiol and IGF-I were quantified in daily plasma samples from d 8 to 16 during the last ovulatory cycle (Cycle -2) and the subsequent anovulatory cycle (Cycle 1). During the last two cycles before anovulation, M heifers had 50% larger (P < .0001) ovulatory follicles than R heifers and 61% greater (P < .0001) growth rate of the ovulatory follicles. There was a treatment x cycle x day effect (P < .001) for concentrations of estradiol. The preovulatory increase in estradiol occurred in the R and M heifers during Cycle -2 but only in M heifers during Cycle -1. A treatment x cycle x day effect (P < .05) influenced LH concentrations. During Cycle -2, LH concentrations were similar for M and R heifers, but during Cycle 1, M heifers had greater LH concentrations than did R heifers. Concentrations of FSH were greater (P < .05) in R than M heifers after induced luteolysis when R heifers failed to ovulate. There was a treatment x cycle interaction (P < .05) for IGF-I concentrations, and M heifers had 4.7- and 8.6-fold greater IGF-I concentrations than did R heifers during Cycle -2 and -1, respectively. We conclude that growth rate and diameter of the ovulatory follicle, and concentrations of LH, estradiol, and IGF-I are reduced before the onset of nutritionally induced anovulation in beef heifers. PMID- 10375233 TI - Adrenocortical response to ACTH in Angora and Spanish goat wethers. AB - Angora goats do not cope well with stress compared with goats of other breeds. Our hypothesis that this involves subclinical primary hypoadrenocorticism associated with low cortisol release in response to ACTH stimulation was tested by measuring adrenocortical response (plasma cortisol) in six Spanish (37 +/- 2 kg BW) and six Angora wethers (39 +/- 3 kg BW) under simulated acute and chronic ACTH challenges. In Exp. 1 (acute ACTH challenge), wethers were dosed i.v. with high (2.5 IU/kg BW) or low (.4 IU/kg BW) quantities of ACTH. In Exp. 2 (chronic ACTH challenge), ACTH at the rate of .015 IU/(kg BW x min) or saline (.15 M NaCl) was infused i.v. at 15 mL/h for 6 h. The mean baseline plasma cortisol concentration before ACTH stimulation was similar (P > .05) between Angora and Spanish goats in Exp. 1 (averaged over days) and in Exp. 2. The cortisol concentration response area (ng/ (mL x min) x 10(-3)) above the baseline was similar (P > .05) between Angora and Spanish goats during low (7.6 +/- .5 and 9.0 +/- 1.7, respectively) and high (12.8 +/- 1.0 and 16.0 +/- 1.8, respectively) levels of acute ACTH challenge (Exp. 1) and during chronic ACTH challenge (45.1 +/- 5.9 and 41.8 +/- 7.3, respectively; Exp. 2). In conclusion, these data indicate that, under the conditions of this study, adrenocortical responsiveness to ACTH stimulation is not different between Angora and Spanish goat wethers and, thus, may not contribute to stress susceptibility in Angora goats. PMID- 10375232 TI - Identification of IGF binding proteins in bovine milk and the demonstration of IGFBP-3 synthesis and release by bovine mammary epithelial cells. AB - Insulin-like growth factor system components are synthesized and secreted by mammary epithelial cells and multiple IGF binding proteins (IGFBP) are found in milk of various species. This study was conducted to identify the IGFBP in bovine milk, to compare them with those found in blood, and to identify the cell(s) responsible for mammary IGFBP synthesis. Bovine blood, milk, and cell culture conditioned media were analyzed and characterized with Western ligand blot procedures for specific IGFBP. Electrophoresis and [125I]IGF-II ligand blot analyses of the samples indicated that, unlike serum and mammary primary cell culture-conditioned media, milk required removal of casein in order to accurately disclose all IGFBP. Immunoprecipitation studies identified IGFBP-2, -3, -4, and 5 in blood, milk, and primary cell culture conditioned media. The IGFBP were present at higher concentrations in serum than in milk, and milk concentrations were greater than that shown in conditioned media from primary cultures of bovine mammary cells. Northern analysis detected IGFBP-3 messenger RNA in extracts from fresh tissue and cells in culture, and in situ hybridization studies with fresh tissue utilizing probes for IGFBP-3 and alphaS1-casein showed that the mRNA for IGFBP-3 is predominant in the secretory epithelial cells, when compared to other tissue cell types. PMID- 10375234 TI - In situ neutral detergent insoluble nitrogen as a method for measuring forage protein degradability. AB - A method of estimating the undegraded intake protein (UIP) concentration of forages was developed and validated with a series of in situ experiments. The hypothesis was that UIP calculated from in situ neutral detergent insoluble N (NDIN) is equal to total in situ N minus the microbial N that is estimated from purines (MN). The in situ disappearance rates of total in situ N (TN), MN, and NDIN were measured for six hay samples and two range masticate samples. Hypothetical rates of passage (2 or 5%/h) were used to calculate UIP (% of DM) for each N pool. Estimates of UIP from TN were higher (P = .0001) than those from either MN or NDIN, and MN estimates of UIP were similar (P = .48) to NDIN estimates. A low-N fiber source (solka floc) was incubated in situ for 8 h. Analysis of the residue detected purines before, but not after, neutral detergent extraction. Several in situ incubation (i.e., Dacron bag size and number of Dacron bags in a mesh bag) and neutral detergent extraction conditions were tested. None of the factors tested affected in situ NDIN disappearance (P > .05). The hypothesis that NDIN is completely digestible in the rumen was tested. Estimates of the extent of NDIN digestion were made using 96-h in situ incubations, and UIP was recalculated for the test samples. Mean in situ UIP concentration decreased upon recalculation (P = .05). In situ NDIN provides estimates of forage UIP that are equal to estimates from MN. Forage UIP estimates are less when extent of N degradation is estimated and included in the calculation. PMID- 10375235 TI - Comparison of Tifton 85 and Coastal bermudagrasses for yield, nutrient traits, intake, and digestion by growing beef steers. AB - A study was undertaken to compare Tifton 85 (T85) and Coastal (CBG) bermudagrasses for effects of cultivar and age at harvest on yields of DM and digestible DM, in vitro digestion, nutrient content, cell wall composition, in situ digestion kinetics, and feed intake and digestion by growing beef steers. In Exp. 1, T85 and CBG forages staged for growth in May or July of 1993 were harvested at 3, 4, 5, 6, 7, and 8 wk from subplots. Tifton 85 bermudagrass had 7.1% greater DM yield, 18.2% higher (P < .05) digestible DM yield, and 7.1% greater IVDMD than CBG, and, after 5 wk of forage growth, IVDMD of both T85 and CBG decreased with increased age at harvest (P < .05). In Exp. 2, T85 and CBG forages staged for growth in July 1997 were harvested at 2, 3, 4, 5, 6, and 7 wk from subplots. Even though T85 had higher concentrations of NDF and ADF than CBG, T85 had 34.1% higher DM yield, 47.9% higher digestible DM, 55.0% higher digestible NDF, 91.7% higher digestible ADF, greater IVDMD, in vitro NDF and ADF disappearances, and higher in situ DM and NDF digestion (P < .05). Coastal bermudagrass had higher concentrations of lignin and lower concentrations of total neutral sugars, arabinose, glucose, and xylose than T85 (P < .05). In vitro digestibilities of DM, NDF, and ADF were lower and concentrations of ADF and lignin were greater for 7- vs 6-wk harvests of both T85 and CBG (P < .05). In Exp. 3, T85 and CBG forages staged for growth in July 1997 were harvested as hay at 3, 5, and 7 wk from .8-ha pastures and fed to 36 individually penned growing beef steers (initial BW = 244 kg) to quantify ad libitum intake without supplementation. Tifton 85 bermudagrass had lower concentrations of lignin and ether-linked ferulic acid and greater concentrations of NDF, ADF, hemicellulose, and cellulose than CBG (P < .05). Steers fed T85 had higher (P < .05) digestion of DM, OM, NDF, ADF, hemicellulose, and cellulose than steers fed CBG. Digestion of NDF, ADF, hemicellulose, and cellulose decreased (P < .05) with increased age at harvest for both cultivars. In conclusion, T85 produced more DM and had more digestible nutrients in vitro, in situ, and in vivo than CBG, and 3 and 5 wk of growth would be recommended ages to harvest either cultivar. PMID- 10375236 TI - Influence of grain source on ruminal characteristics and rate, site, and extent of digestion in beef steers. AB - Six cannulated Salers steers (305 +/- 17 kg initial BW) were used in a double 3 x 3 Latin square design to compare the effects of the nature of the cereal (wheat vs corn) and the corn genotype (dent vs flint) on rate, site, and extent of digestion of high-concentrate diets. The cereals were coarsely cracked, and the diets were balanced to have the same percentage of starch (47.7 +/- 2.3%) and CP (14.6 +/- .7%). Differences in ruminal starch digestion were observed between wheat- and corn-based diets (86.6 vs 47.8%; P < .001) and between corn genotypes (60.8 vs 34.8% for dent and flint corns; P < .001). For flint corn, more than half the starch was digested in the hindgut. Total tract digestion of starch was greater (P < .001) by steers fed wheat than by those fed corn and did not differ (P > .1) between the two corn genotypes. Ruminal mean pH (P < .01) was lower and total VFA concentration (P < .1) was higher for wheat- than for corn-based diets. Ruminal acetate:propionate tended to increase with the decrease in the amount of starch degraded in the rumen, but differences were not significant (P > .1). When wheat replaced corn, nonammonia, nonmicrobial N duodenal flow decreased (P < .01), and microbial duodenal flow increased (P < .05), so there were no differences in the duodenal flow of nonammonia N duodenal flow (P > .1). The lower nonammonia N duodenal flow for the dent corn- than for the flint corn-based diet (P < .05) was related to a lower passage of nonammonia, nonmicrobial N into the duodenum. Efficiency of microbial protein synthesis was inversely correlated with the amount of starch degraded in the rumen. Nature of the cereal, wheat vs corn, and genotype of the corn, dent vs flint, alter the site and extent of starch digestion. PMID- 10375237 TI - Effectiveness of calcium chloride in increasing blood calcium concentrations of periparturient dairy cows. AB - Calcium chloride supplements such as gels and drench were studied to determine their effectiveness for increasing blood serum Ca concentrations in periparturient dairy cows. Multiparous, pregnant Holstein dairy cows (n = 36) were assigned to one of four treatments. After calving, cows in four treatments received basal diet and two doses of either control inert gel (CON), gel containing CaCl2 and vitamins (CVG), gel containing CaCl2 and minerals (CMG), or CaCl2 as drench containing vitamins (CVD). The first dose was given within 2 h after calving and the second dose 12 h after the first dose. Each dose provided .07, 54.5, 56.0, and 33.2 g of elemental Ca in CON, CVG, CMG, and CVD treatments, respectively. Blood samples were collected at 0, 15, 30, 60, 180, and 360 min after each oral dose. The blood serum Ca concentrations were 6.26, 7.56, 6.20, and 5.96 mg/dL during the pretreatment period and deviated -13.5, 7.1, 9.3, and 18.1% from pretreatment levels at 18 h after the first dose in CON, CVG, CMG, and CVD treatments, respectively. The average changes in serum P from pretreatment levels were not different among treatments. Serum Mg concentrations remained below the pretreatment levels in all four treatments. Blood serum beta hydroxybutyrate during the first 2 wk and milk yields during the first 4 wk of lactation were the same in all treatments. Three cases of clinical milk fever were observed in CON treatment and one case in CVD treatment. The oral supplements of CaCl2 as gel or drench increased the blood Ca levels in periparturient dairy cows. Increased supply of Ca through oral supplements of CaCl2 may prevent milk fever in cows that are marginally hypocalcemic. PMID- 10375238 TI - Myopia: an epidemic of possibilities? PMID- 10375239 TI - Factors associated with the prevalence of myopia in 6-year-olds. AB - PURPOSE: To describe the frequency of and risk factors associated with myopia in grade one children. METHODS: Refractive error was measured by static retinoscopy, without cycloplegia, for 10,616 children in the first year of a province-wide vision-screening program. Information on factors that might be associated with myopia was collected from parents or guardians by self-administered questionnaires distributed before the vision screening. These factors were evaluated by a case-control method. RESULTS: The prevalence of myopia, greater than -0.25 D, was 6%. The estimated relative risk of myopia was increased significantly among children whose birth weight was <2500 g and whose mothers had a history of early spectacle use. CONCLUSIONS: Results suggest that the prevalence of myopia in 6-year-old children is associated with both hereditary and nonhereditary factors. In accord with prior work, the results argue that low birth weight has a permanent influence upon eye development. PMID- 10375240 TI - Is refraction in early infancy a predictor of myopia at the age of 7 to 8 years? The relationship between cycloplegic refraction at 11 weeks and the manifest refraction at age 7 to 8 years in Chinese children. AB - Thirty-two children who had been refracted in early infancy were re-refracted at the age of 7 to 8 years. Refraction comprised cycloplegic retinoscopy at the mean age of 11 weeks and noncycloplegic retinoscopy and subjective examination at the mean age of 94 months. The change in spherical equivalent refraction (SER) was highly negatively correlated with the initial refraction (-0.863), demonstrating emmetropization; however, the correlation between the refractive error at age 7 to 8 years and the refractive error in infancy was much weaker (+0.225). The SER was significantly less and the astigmatism was greater in infancy in children who were myopic at age 7 to 8 years. There was, however, extensive overlap in range between the SER for the two groups and the initial SER was not a good predictor of myopia at 7 to 8 years, although it may help to identify children who are unlikely to become myopic. One hyperopic child with bilateral ptosis failed to emmetropize. PMID- 10375241 TI - Epidemiologic study of ocular refraction among schoolchildren in Taiwan in 1995. AB - PURPOSE: In order to understand and update the prevalence of myopia in Taiwan, a nationwide survey was performed in 1995. METHODS: We stratified the cluster sampling by developmental grading of the city, using a size proportional to the population. Two cities were randomly selected from each city grading. The total number of students enrolled was 11,178, including 5,676 boys and 5,502 girls. The refractive status and corneal radius of each student were measured with an autorefractometer under cycloplegia and checked with retinoscopy. Axial length was measured with biometric ultrasound. RESULTS: The myopic rate was from 12% at the age of 6, it increased to 56% at the age of 12, and then to 76% at the age of 15. A myopic rate of 84% was found for the age range of 16 to 18. The prevalence of high myopia (over -6.0 D) at the age of 18 was 20% in girls and 12% in boys. The mean refractive status became myopic at the age of 9, then increased to -3.92 D in girls and -2.71 D in boys at the age of 18. The increase of axial length is correspondent with the progression of myopia. The anterior chamber depth (ACD) was deeper with age and the severity of myopia, whereas the corneal curvature remained unchanged. The lens thickness became thinner from age 7 to 13, then it became thicker with age and the severity of myopia after age 15. The prevalence and degree of myopia in girls was more severe than in boys. CONCLUSIONS: The prevalence of myopia in Taiwan increased year by year. The increase in severity and prevalence of high myopia may be due to earlier onset. PMID- 10375242 TI - Prevalence of myopia in Sherpa and Tibetan children in Nepal. AB - BACKGROUND: Tibetan and Sherpa children living in Nepal share a common ancestry in Tibet and areas to the north of Nepal, but it is evident that these people experience widely contrasting educational and environmental conditions. The purpose of this study was to compare the prevalence of myopia in children with similar genetic backgrounds but who are exposed to contrasting environments. METHODS: Unaided vision and refractive error was measured in 555 Tibetan children in Kathmandu and 270 Sherpa children in the Solu Khumbu region of Nepal. RESULTS: There were marked differences in vision and the prevalence of myopia in the two groups. Ninety-two percent of the Sherpa children had Snellen vision of 20/22 (0.89) or better compared with 70% of the Tibetan children. The range of refractive errors was -6.50 to +7.00 D for the Tibetan children and -1.00 to +3.50 D for the Sherpa children. The Sherpa children had a prevalence of myopia of 2.9% compared with 21.7% for the Tibetan children. CONCLUSIONS: The prevalence of myopia in Sherpa children is low and their rural lifestyle appears to be relatively unstressed. Tibetan children have a higher prevalence of myopia and more rigorous schooling. We did not establish a causal relationship between myopia and the type of schooling, or the environment in general, but we did demonstrate that a simple, rural lifestyle is at least compatible with a virtual absence of myopia. PMID- 10375243 TI - Clinical findings before the onset of myopia in youth: 5. Intraocular pressure. AB - BACKGROUND: We conducted a study to compare variables in children who were initially emmetropic and subsequently became myopic to those in children who remained emmetropic. This paper investigates intraocular pressure (IOP) in the two groups. Some theories of myopia development suggest a role for elevated IOP in axial elongation of the eye, and some studies have found higher IOPs in myopes than in nonmyopes. METHODS: A cohort of initially emmetropic children were given eye and vision examinations at 6-month intervals over a 3-year period. IOPs were obtained by Goldmann tonometry. RESULTS: The mean IOP for the became-myopic group was 13.9 mm Hg (N = 24; SD = 2.5). The mean IOP for the remained-emmetropic group was 14.7 mm Hg (N = 53; SD = 3.1). The difference was not statistically significant. CONCLUSION: Children who became myopic did not show a greater IOP at an examination 6 months or less before the onset of myopia than children who remained emmetropic. PMID- 10375244 TI - Relationship of accommodative response and nearpoint phoria in a sample of myopic children. AB - Esophoria has been associated with onset and progression of myopia in children. The induction of myopia by optical defocus shown by animal models suggests that a high lag of accommodation during near work may contribute to myopia in children. This paper examines the relationship of nearpoint phoria and accommodative response in a sample of children with myopia. Accommodative response was measured under binocular conditions with the Canon Autoref R-1 autorefractor with a 40 cm viewing distance. Phorias were measured with the von Graefe method using a 40 cm test distance. In the statistical analysis exophoria was scored as a negative number and esophoria was scored as a positive number. The coefficient of correlation of accommodative response with phoria was -0.32 (n = 73; p<0.01), thus showing an association of a more positive (more convergent) near phoria with lower accommodative response. The correlation coefficient increased to -0.39 when an exponential function was used. When only esophores were considered, the correlation coefficient was -0.59 (n = 44; p<0.001). Lower accommodative response (higher lag of accommodation) was associated with greater esophoria. PMID- 10375245 TI - Effect of accommodative adaptation on static and dynamic accommodation in emmetropia and late-onset myopia. AB - PURPOSE: Prolonged near work is considered to be an environmental factor leading to the development of late-onset myopia. Accommodation may be the specific mechanism underlying the association between near work and late-onset myopia. To determine whether late-onset myopes have abnormal accommodation, we compared accommodative static responses and dynamic facilities before and after a near task in two groups of subjects, emmetropes and late-onset myopes. METHODS: In experiment 1, the accommodative stimulus/response function with monocular viewing and the dark focus (the accommodative response in the dark) were objectively measured with a Canon R-1 infrared optometer before (preadaptation) and after (postadaptation) a 20-min near task. In experiment 2, monocular accommodative facility (AF) and dark focus were measured before and after the near task. Facility was measured as the subjective time needed to clear an accommodative target (20/40 letters) at 40 cm through +/-2.00 D lenses. The time between when the subject flipped the lenses from viewing through the +2.00 D to the -2.00 D lenses was recorded by a computer. RESULTS: In both experiments, inward shifts of the dark focus were observed after the near task. In experiment 1, after the near task, static accommodative responses also showed a small but statistically significant inward shift. Neither postadaptation effect differed between refractive groups. The only difference between groups was that late-onset myopes had a lower slope of the accommodative stimulus/response function, both pre- and postadaptation. In experiment 2, both refractive groups showed the same results. After the near task, the duration for accommodation from near to far (relaxation) increased but the duration for accommodation from far to near (stimulation) did not change. CONCLUSIONS: Late-onset myopes have shallower accommodative stimulus/response functions. As suggested in a previous study, this may be due to their reduced sensitivity to defocus. In both emmetropes and late-onset myopes, the near task causes an increase in static accommodative responses. Although our results show it to be a small increase, it is consistent with predictions of Hung and Semmlow's model of accommodation. In both emmetropes and late-onset myopes, the near task also increases the duration for relaxing accommodation, but not for stimulating accommodation. This suggests there are two subsystems which may adapt to prolonged accommodation differently. PMID- 10375246 TI - Blur sensitivity in myopes. AB - PURPOSE: This study compared the ability of myopes and emmetropes to detect subjectively the presence of retinal defocus. METHODS: Subjects (12 myopes, 12 emmetropes) were cyclopleged and monocularly viewed a bipartite target through an appropriate near addition lens via a 2-mm artificial pupil. One-half of the target remained fixed while the other half was alternatively moved forward or backward until subjects first reported a difference in clarity between the two halves of the target. RESULTS: The mean blur threshold for the emmetropes and myopes was +/-0.11 and +/-0.19 D, respectively (p = 0.0001). CONCLUSIONS: These results demonstrate that myopes are less sensitive to the presence of blur, and may at least partially explain why previous reports have demonstrated a larger lag of accommodation in this refractive group. Additionally, the hyperopic retinal defocus resulting from the increased accommodative error may play a significant role in myopia development and progression. PMID- 10375247 TI - Effect of interrupted lens wear on compensation for a minus lens in tree shrews. AB - BACKGROUND: When a young animal wears a monocular minus (concave) lens that shifts the focal plane away from the cornea, the vitreous chamber elongates over a period of days, shifting the retinal location to compensate for the altered focal plane. We examined the effect of removing the lens for a portion of each day on the amount of compensation in tree shrews. METHODS: Starting 24 days after natural eye opening, juvenile tree shrews wore a goggle frame that held a -5 D lens in front of one eye, with an open frame around the fellow control eye. The goggle was removed for 0, 0.5, 1, 2, or 7 h each day (N = 5, 5, 5, 5, and 3 animals per group, respectively), starting 0.5 h after the start of each 14 h light-on period. After 21 days of treatment, measures were made of the cycloplegic refractive state (streak retinoscopy) and the ocular component dimensions (A-scan ultrasound). Normal animals that experienced 14 h each day with no lens (N = 3) were also examined. RESULTS: The treated eyes of the 0 h group developed full refractive compensation for the lens (treated eye - control eye, mean +/- SEM = -5.8+/-1.1 D) and had increased vitreous chamber depth (0.13+/-0.02 mm) and axial length (0.12+/-0.02 mm) relative to the untreated control eye. The groups in which the lens was removed for 0.5 and 1 h each day showed partial compensation for the -5 D lens, both in refractive state (-4.2+/ 0.4 D; -2.9+/-1.6 D) and in vitreous chamber depth (0.12+/-0.02 mm; 0.09+/-0.02 mm). The 2, 7, and 14 h (normal) groups showed no significant refractive or axial compensation. In the 0.5 and 1 h groups, A-scan ultrasound showed a thinning of the region between the front of the retina and back of the sclera. CONCLUSIONS: The eyes of tree shrews can tolerate altered monocular visual stimulation produced by a minus lens worn for 12 h of a 14-h light cycle without developing an induced myopia. However, when the lens is worn more than 12 of 14 h each day, compensation appears to increase linearly with decreased lens-off time. If the eyes of human children respond similarly to defocus from near work or other sources, it would seem that the defocus must be present almost all the time to induce myopia. If defocus contributes to human myopia through a compensation mechanism, then an increase in the amount of time that focused images are present should reduce myopic progression. PMID- 10375248 TI - Afocal magnification does not influence chick eye development. AB - In defocus-induced ametropia experiments, retinal blur circles are a likely source of information as to the magnitude but not the sign of the defocus. However, magnification (and minification) produced by the lenses may be a cue. In this study, 1-day-old broiler chicks (N = 13) were treated monocularly for 7 days with special goggles containing approximately afocal iseikonic lenses which were designed to produce 10% retinal image magnification. This is a little less than the magnification produced by +10 D defocusing lenses used to produce about 10 D of hyperopia in earlier work. Intraocular dimensions of both eyes were measured by A-scan ultrasonography on the first and last day. Refractive states of both eyes were measured daily with a retinoscope and trial lenses. After the birds were sacrificed, the eyes were enucleated, weighed, and measured with calipers. Before the treatment there was no difference in the refractive state or dimensions of the right and left eyes. After 1 week of goggle wear there was still no significant difference between the eyes in spite of the magnification produced by the goggles. These data suggest that factors other than magnification are responsible for the ability of the eye to respond to the sign of defocus. PMID- 10375249 TI - Imposed retinal image size changes--do they provide a cue to the sign of lens induced defocus in chick? AB - BACKGROUND: Young chicks can adjust their eye growth to compensate for both imposed hyperopia and myopia (using negative and positive spectacle lenses); the rate of eye elongation increases in the former and slows in the latter case. This emmetropizing behavior implies that the eye can distinguish the sign and magnitude of defocus, although the identity of the cue(s) involved is unknown. As the spectacle lenses used in these studies generally introduce significant retinal image size differences that are in opposite directions for negative and positive lenses (minification vs. magnification), we asked whether retinal image size might provide the required sign information. METHODS: This question was addressed by manipulating retinal image size while keeping lens power constant. We also investigated the effect of eliminating other potential cues, accommodation and chromatic aberration, under these conditions. Three negative "size" lenses of approximately -11 D optical power were used, with 2 of the lenses producing magnification rather than minification as typical of negative lenses (i.e. +1.9% and +6.9% compared to -2.9%). The lenses were fitted monocularly to 7-day-old chicks, which were subsequently measured at 9 and 11 days of age (refractive error and axial dimensions). The same lens-wearing schedule was applied to two other groups of chicks that had monocular ciliary nerve section surgery to prevent accommodation 2 days posthatching; one of these groups was reared under monochromatic yellow light instead of white light. RESULTS: Near-perfect refractive compensation was seen by the end of the treatment period with all three lenses, for all three treatment groups, and there was also little difference in the rate of compensation among the various groups. In all cases, the typical responses of axial (mainly vitreous chamber) elongation and myopia were observed. CONCLUSIONS: That manipulations to retinal image size, which either decrease or reverse the usual effects of negative lenses, did not disrupt compensation to the imposed hyperopic defocus, even in the absence of accommodation and chromatic aberration cues, argues against imposed retinal image size changes being the directional cue to defocus in experimental emmetropization. PMID- 10375250 TI - Cone receptor sensitivity is altered in form deprivation myopia in the chicken. AB - PURPOSE: The effect of form deprivation myopia (FDM) on cone photoreceptor function was investigated. METHODS: Photopic electroretinogram (ERG) a-waves were recorded from control and form-deprived eyes of chickens. Cone-generated P3 responses were derived from the leading edge of ERG a-waves using an analytical expression derived from a quantitative model of phototransduction. The parameters obtained were the maximum cone P3 response (RmaxP3), sensitivity (S), and delay (t(d)). RESULTS: P3 response sensitivity is significantly higher in form-deprived eyes, at lower retinal irradiances. At higher flash intensities, in form-deprived eyes, P3 response sensitivity declines at a significantly greater rate, per unit increase in retinal irradiance, than in control eyes. Visual deprivation does not significantly affect RmaxP3 or t(d). CONCLUSIONS: Hypotheses to explain the altered cone photoreceptor sensitivity in form-deprived eyes are proposed. Changes in the biochemistry of phototransduction, or intrinsic geometric and physical attributes of photoreceptors or their waveguide modal properties, could account for the findings. PMID- 10375251 TI - Current awareness in NMR in biomedicine. PMID- 10375252 TI - International Cancer Gene Therapy Symposium. London, England, May 28-29, 1998. Abstracts. PMID- 10375253 TI - Proceedings of the Vth International Workshop on Cytokines. Florence, Italy, March 16-18, 1998. PMID- 10375254 TI - Psoriasis--practical solutions to difficult patients. PMID- 10375255 TI - [Congenital orbital diseases]. PMID- 10375256 TI - Mechanism of methacholine dose-response plateus in normal subjects. PMID- 10375257 TI - [Occupational Medicine Institute of the Academy of Medical Sciences of Ukraine: history, present, achievement and perspectives (for 70th anniversary of its foundation)]. PMID- 10375259 TI - 20th Annual meeting of the Japanese Society of Biological Psychiatry and 15th meeting of the Japanese Association for Adolescent Psychiatry. Abstracts. PMID- 10375258 TI - Proceedings of the 8th Swiss Workshop of Methodology in Receptor Research. Morschach, May 3-7, 1998. PMID- 10375260 TI - Retraction. PMID- 10375261 TI - HIV laws: a new shift in focus. PMID- 10375262 TI - Megestrol acetate: promises and pitfalls. AB - Recent reports suggest that effective antiretroviral therapy, resulting in a plasma HIV load that has been reduced to undetectable levels, may itself prevent HIV- and opportunistic infection-associated weight loss and lead to substantial weight gain. Although these data are encouraging, it is clear that a significant proportion of patients will require, in addition, specific treatment for HIV associated wasting. Megestrol acetate, in the dosage range of 400 to 800 mg/day, is a useful appetite stimulant for the prevention and treatment of HIV-associated wasting, particularly in women. Patients need to be advised of possible adverse effects and monitored closely. Megestrol acetate stimulates weight gain mostly through an increase in body fat and is therefore most effective in combination with a muscle-building exercise program, where appropriate, and an anabolic agent (steroid or growth hormone) to maintain or increase lean body mass. PMID- 10375263 TI - HIV-associated dementia: new insights into disease pathogenesis and therapeutic interventions. AB - Remarkable progress was made in recent years in the therapeutics of HIV-1 associated dementia (HAD) and in unraveling the complex pathophysiology that follows viral invasion of the central nervous system (CNS). Viral replication in and outside of the CNS was significantly reduced in HIV-1 infected subjects by new potent antiretroviral therapies. This has resulted in partial repair of cellular immune function with improvement in, and the prevention of, neurologic deficits associated with progressive HIV-1 disease. In regard to HAD pathophysiology, it is now known that CNS damage induced by HIV-1 infection occurs indirectly. Neuronal loss is mediated through immune activation and viral infection of mononuclear phagocytes (MPs) (brain macrophages and microglia). Cellular and viral factors secreted by brain MPs produce, over time, neuronal damage and drop out. Viral growth in the brain appears necessary, but not sufficient, to produce cognitive and motor impairments in affected individuals. Indeed, the best predictor for neurologic impairment following HIV-1 infection is the absolute number of immune-competent macrophages; not the level of viral production in affected brain tissue. As yet, an understanding of macrophage related neurodegeneration has not translated into significant improvements in the treatment of this devastating complication of HIV disease. Nonetheless, adjunctive antiinflammatory and neuroprotective therapies are being developed. New ideas regarding HAD neuropathogenesis, and implications for the diagnosis and treatment of HAD are summarized in this article. PMID- 10375264 TI - HIV/AIDS rounds: HIV in children. PMID- 10375265 TI - Differences in curable STDs between HIV and non-HIV populations in Spain. AB - The pandemic impact of HIV has changed the clinical spectrum of STDs all over the world. The incidence and frequency of STDs in the different global geographic areas demonstrate the diagnostic and treatment capabilities of various local and national health systems and is simultaneously informing about the sexual behaviours of the population. The purpose of this study was to determine the frequency of curable STDs (herpes, chlamydia, gonorrhoea, syphilis, trichomoniasis) in a hospital-based STD clinic in Madrid, Spain during a 4-year period. Patients were referred mainly from the emergency department, gynecological wards, and family planning (61%) as well as from the HIV-hospital unit (31 beds) and outpatient department (39%). The total number of patients seen was 952 (243 men, 709 women) with an annual average of 238 patients per year. Of these, 139 (14.6%) were HIV-patients and 813 (85.4%) non-HIV patients. In non-HIV patients, STDs were identified in 493 cases (54.2%). In HIV-patients, STDs were diagnosed in 108 cases (77.7%; p < or = 0.001). Two or more STDs were more prevalent in HIV than non-HIV patients. The frequency of STDs in both HIV and non HIV patients were vulvovaginal candidiasis, 47.8%:57.2%; syphilis, 11.7%:1.4% (p < or = 0.05); gonorrhea, 5.3%:3.9%; Gardnerella vaginosis, 6.3%:4.8%; genital chlamydia, 6.3%:9.06%; trichomoniasis, 17%:6.5% (p < or = 0.05); and genital herpes, 20.2%:5.3% (p < or = 0.05). PMID- 10375266 TI - Prevalence of enteric pathogens in HIV-related diarrhea in the midwest. AB - HIV disease is often associated with the condition of diarrhea, which may be accompanied by enteric infection or gastrointestinal tumor. This study prospectively investigated 27 episodes of chronic diarrhea in 24 patients with HIV infection. Upper endoscopy and sigmoidoscopy with biopsies at three sites (distal duodenum, sigmoid colon, and rectum) and viral and mycobacterial blood cultures were performed. Stool specimens were sent for standard tests. A primary infectious diagnosis was found in 10 (37%) of 27 episodes: cytomegalovirus (CMV) colitis (n = 4), 3 microsporidiosis (n = 3), cryptosporidiosis (n = 2), and colonic histoplasmosis (n = 1). Patients with CD4 counts of less than 50 cells/mm3 and with lower albumin levels were more likely to have a primary infectious diagnosis. Adenovirus was found in 7 cases but was often associated with another organism; these were not considered to be primary diagnoses. Blood cultures for viruses were not useful, and all mycobacterial cultures were negative. A flexible sigmoidoscopy with histologic examination and culture of biopsy samples were the diagnostic tools that yielded most infectious diagnoses. Follow-up showed that two thirds of patients improved with nonspecific antidiarrheal medications regardless of diagnosis. The study supports a minimalistic approach to the problem of diarrhea in patients with HIV infection. Upper and lower endoscopy lead to a precise diagnosis in a minority of cases, and the outcome was similar in patients with or without a primary infectious diagnosis. PMID- 10375267 TI - Predictors of self-reported adherence in persons living with HIV disease. AB - This study examined the relationships between the five dimensions of the Wilson and Cleary model of health-related quality of life and three self-reported adherence measures in persons living with HIV using a descriptive survey design. Data collection occurred in seven cities across the United States, including university-based AIDS clinics, private practices, public and for-profit hospitals, residential and day-care facilities, community-based organizations, and home care. The three dependent adherence measures studied were "medication nonadherence," "follows provider advice," and "missed appointments." The sample included 420 persons living with HIV disease with a mean age of 39 years of which 20% were women and 51% were white; subjects had a mean CD4 count of 321 mm3. HIV positive clients with higher symptom scores, particularly depression, were more likely to be nonadherent to medication, not to follow provider advice, and to miss appointments. Participants who reported having a meaningful life, feeling comfortable and well cared for, using their time wisely, and taking time for important things were both more adherent to their medications and more likely to follow provider's advice. No evidence was found demonstrating any relationship between adherence and age, gender, ethnicity, or history of injection drug use. These findings support the need to treat symptoms, particularly depression, and to understand clients' perceptions of their environment as strategies to enhance adherence. A limitation of this study was that adherence was measured only by self-report; however, the study did expand the concept of adherence in HIV care beyond medication adherence to include following instructions and keeping appointments. PMID- 10375268 TI - ABT-378 lowers viral load. PMID- 10375269 TI - Directly observed HAART. PMID- 10375271 TI - Baltimore highest in STDs. PMID- 10375270 TI - HBV resistance in HIV patients. PMID- 10375272 TI - STDs may cause early deliveries. PMID- 10375273 TI - STD myths among teen girls. PMID- 10375274 TI - CDC ranks cities and STDs. PMID- 10375275 TI - Trial will evaluate cyclosporine. PMID- 10375276 TI - Impact of individualized counseling on HIV infection rates. PMID- 10375277 TI - Diarrhea-associated diffusely adherent Escherichia coli. PMID- 10375278 TI - World asthma meeting, Barcelona, Spain, December 1998. PMID- 10375280 TI - Challenges in Cancer Metastasis. Proceedings of the 14th Bristol-Myers Squibb Nagoya International Cancer Treatment Symposium. Nagoya, Japan, 11-12 September 1998. PMID- 10375279 TI - Percutaneous ultrasound-guided radiofrequency ablation of liver tumors. PMID- 10375281 TI - Reduction of paraphimosis with granulated sugar. PMID- 10375282 TI - Urinary incontinence: an unusual manifestation of a forgotten stent. PMID- 10375283 TI - Urodynamic variable cannot be used to classify the severity of detrusor instability. PMID- 10375284 TI - Urodynamic variables cannot be used to classify the severity of detrusor instability. PMID- 10375285 TI - Periurethral haematomas in a child following urethral dilation for dysfunctional voiding. PMID- 10375287 TI - Wombat uroflowmetry. PMID- 10375286 TI - The use of the pusher to prevent intracystoscopic displacement of a 'backloaded' guidewire. PMID- 10375288 TI - Physician strategies for the 21st century. PMID- 10375289 TI - Therapeutic approaches to type 2 diabetes. Proceedings of a conference. Scottsdale, Arizona, USA. 10-12 December 1997. PMID- 10375290 TI - Insulin resistance and hyperglycemic associated risk factors: session summary. PMID- 10375291 TI - Impact of glucose control in type 2 diabetes: session summary. PMID- 10375292 TI - Overview of current therapeutic options: session summary. PMID- 10375294 TI - Genes and Gynecology. Proceedings of the 2nd Symposium of the Wim Schellekens Foundation. The Hague, The Netherlands, June 12, 1998. PMID- 10375293 TI - Buildings operations and ETS exposure. AB - Mechanical systems are used in buildings to provide conditioned air, dissipate thermal loads, dilute contaminants, and maintain pressure differences. The characteristics of these systems and their operations h implications for the exposures of workers to environmental tobacco smoke (ETS) and for the control of these exposures. This review describes the general features of building ventilation systems and the efficacy of ventilation for controlling contaminant concentrations. Ventilation can reduce the concentration of ETS through dilution, but central heating, ventilating, and air conditioning (HVAC) can also move air throughout a building that has been contaminated by ETS. An understanding of HVAC systems is needed to develop models for exposures of workers to ETS. PMID- 10375295 TI - Alendronate increased bone mineral density but did not reduce new fractures in glucocorticoid induced osteoporosis. PMID- 10375296 TI - A requiem for the cholecystokinin provocation test? PMID- 10375297 TI - Measles virus and Crohn's disease. PMID- 10375298 TI - A UK training programme for nurse practitioner flexible sigmoidoscopy. PMID- 10375300 TI - Cord blood banking. PMID- 10375299 TI - Gas and liquid reflux during transient lower oesophageal sphincter relaxation. PMID- 10375301 TI - Introducing Brian Boycott and Heinz Wassle, the 1999 recipients of the proctor medal. PMID- 10375302 TI - Issue dedicated to Karl Z. Morgan. PMID- 10375303 TI - Hypoxia-inducible factor 1alpha (HIF-1alpha) is a non-heme iron protein. Implications for oxygen sensing. PMID- 10375304 TI - A nomenclature solution to mouse MHC confusion. PMID- 10375305 TI - Genetic variation among 129 substrains: practical consequences. PMID- 10375306 TI - 15th International Leprosy Congress. Beijing, China, 7-12 September 1998. Proceedings and abstracts. PMID- 10375307 TI - Pediatric nephrology in Lebanon. From renal biopsy to kidney transplantation. PMID- 10375308 TI - Managed care in Latin America. PMID- 10375309 TI - Increasing probability of mother-to-child transmission of HHV-8 with increasing maternal antibody titer for HHV-8. PMID- 10375310 TI - An unusual cluster of cases of Castleman's disease during highly active antiretroviral therapy for AIDS. PMID- 10375311 TI - Dietary fiber and colorectal cancer. PMID- 10375312 TI - Dietary fiber and colorectal cancer. PMID- 10375313 TI - Dietary fiber and colorectal cancer. PMID- 10375314 TI - Dietary fiber and colorectal cancer. PMID- 10375315 TI - Dietary fiber and colorectal cancer. PMID- 10375316 TI - Dietary fiber and colorectal cancer. PMID- 10375317 TI - Prevention of cisplatin-induced emesis by a neurokinin-1-receptor antagonist. PMID- 10375318 TI - Atherosclerosis--an inflammatory disease. PMID- 10375319 TI - Atherosclerosis--an inflammatory disease. PMID- 10375320 TI - Atherosclerosis--an inflammatory disease. PMID- 10375321 TI - A baby with a Babinski reflex. PMID- 10375322 TI - Envenomations by rattlesnakes thought to be dead. PMID- 10375323 TI - Case study: management of lymphedema. PMID- 10375324 TI - Pediatric Emergency Medicine Fellowship Programs. PMID- 10375325 TI - [Antidepressive therapy for smoking cessation]. PMID- 10375326 TI - [What is the cost and effectiveness of breast cancer screening in women under 50 years of age?]. PMID- 10375327 TI - Airway secretion and asthma. PMID- 10375328 TI - Mechanisms of Toxicity and Carcinogenesis. Proceedings of the 13th Aspen Cancer Conference. Aspen, Colorado, USA. July 18-21, 1998. PMID- 10375329 TI - [Primary tensile strength of the tendon of the supraspinatus muscle in the human- a biomechanical study]. PMID- 10375330 TI - Omnicompetent graduates. PMID- 10375331 TI - Surgical sketch on Wirsung. PMID- 10375332 TI - [3rd Hans Kehr Symposium for Interdisciplinary Gastroenterology and Visceral Surgery. Oberhof, 15-16 January 1999. Abstracts]. PMID- 10375333 TI - [Prof. Khaim Iosifovich Garkavi (on the centenary of his birth)]. PMID- 10375334 TI - The Coxeter Boyle Gas Anaesthesia Apparatus. PMID- 10375335 TI - Genetic and environmental influences on total-body and central abdominal fat: the effect of physical activity in female twins. AB - BACKGROUND: The increasing prevalence of obesity has focused attention on the contribution of physical activity and its interaction with predisposing genetic factors. OBJECTIVE: To examine 1) the relation between physical activity and total-body and central abdominal fat, independent of genetic and other environmental factors, and 2) the influence of physical activity in persons who are genetically susceptible to generalized or central adiposity. DESIGN: Cross sectional study. SETTING: A London academic teaching hospital. PATIENTS: 970 healthy female twins (mean age, 55.5 years [range, 39 to 70 years]; body mass index, 24.4 kg/m2 [range, 16.4 to 44.0 kg/mg2]). There were 241 monozygotic pairs, 228 dizygotic pairs, and 32 women whose co-twin lacked complete data. Fifty-six percent of participants were of normal weight, 30% were overweight, 7% were obese, and 7% were underweight. MEASUREMENTS: Total-body and central abdominal fat were measured by dual-energy x-ray absorptiometry. Physical activity was assessed by quantitative and semiquantitative questionnaires. Data on dietary intake, socioeconomic status, smoking status, and use of hormone replacement therapy (HRT) were also gathered. RESULTS: Total-body and abdominal central adiposity were lower with higher levels of home, sporting, and sweating associated activity. Total-body and central abdominal fat were 5.6 kg and 0.44 kg lower, respectively, in participants who reported vigorous weight-bearing activity. Physical activity was the strongest independent predictor of total-body fat (beta = -0.6 [CI, -1.06 to -0.15]; P = 0.009) and central abdominal fat (beta = -0.07 [CI, -0.1 to -0.03]; P < 0.001) in a regression model that included age, diet, smoking, HRT use, and socioeconomic status. Monozygotic twin pairs who were concordant for smoking and HRT status but were discordant for moderate-intensity sport showed greater within-pair differences in total-body fat than those who were concordant for activity level. In this model, 1 and 2 hours of moderate intensity sport accounted for within-pair differences of 1.0 kg (P = 0.050) and 1.4 kg (P = 0.040), respectively, of total-body fat. In participants who had an overweight twin, higher levels of physical activity were still associated with 3.96-kg lower total-body fat and 0.53-kg lower central abdominal fat. CONCLUSIONS: Current physical activity predicts lower total-body and central abdominal adiposity in healthy middle-aged women. After controlling for genetic and environmental factors, the influence of physical activity was greater than that of other measured environmental factors. Participants with a genetic predisposition to adiposity did not show a lesser effect of physical activity on body fat mass. PMID- 10375336 TI - Association between body mass and adenocarcinoma of the esophagus and gastric cardia. AB - BACKGROUND: The incidence of esophageal and gastric cardia adenocarcinoma is, for unknown reasons, increasing dramatically. A weak and inconsistent association between body mass index (BMI) and adenocarcinoma of the esophagus and gastric cardia has been reported. OBJECTIVE: To reexamine the association between BMI and development of adenocarcinoma of the esophagus and gastric cardia. DESIGN: Nationwide, population-based case-control study. SETTING: Sweden, 1995 through 1997. PATIENTS: Patients younger than 80 years of age who had recently received a diagnosis were eligible. Comprehensive organization ensured rapid case ascertainment. Controls were randomly selected from the continuously updated population register. Interviews were conducted with 189 patients with adenocarcinoma of the esophagus and 262 patients with adenocarcinoma of the gastric cardia; for comparison, 167 patients with incident esophageal squamous cell carcinoma and 820 controls were also interviewed. MEASUREMENTS: Odds ratios were determined from BMI and cancer case-control status. Odds ratios estimated the relative risk for the two adenocarcinomas studied and were calculated by multivariate logistic regression with adjustment for potential confounding factors. RESULTS: A strong dose-dependent relation existed between BMI and esophageal adenocarcinoma. The adjusted odds ratio was 7.6 (95% CI, 3.8 to 15.2) among persons in the highest BMI quartile compared with persons in the lowest. Obese persons (persons with a BMI > 30 kg/m2) had an odds ratio of 16.2 (CI, 6.3 to 41.4) compared with the leanest persons (persons with a BMI < 22 kg/m2). The odds ratio for patients with cardia adenocarcinoma was 2.3 (CI, 1.5 to 3.6) in those in the highest BMI quartile compared with those in the lowest BMI quartile and 4.3 (CI, 2.1 to 8.7) among obese persons. Esophageal squamous-cell carcinoma was not associated with BMI. CONCLUSIONS: The association between BMI and esophageal adenocarcinoma is strong and is not explained by bias or confounding. The carcinogenic mechanism, however, remains to be clarified. The increasing prevalence of obesity in western countries could be important in understanding the increasing occurrence of this tumor. PMID- 10375337 TI - Large pleural effusions occurring after coronary artery bypass grafting. Cardiovascular Surgery Associates, PC. AB - BACKGROUND: Large pleural effusions sometimes occur after coronary artery bypass grafting (CABG), but their characteristics and clinical course are largely unknown. OBJECTIVE: To describe the clinical course and pleural fluid findings in patients with large pleural effusions occurring after CABG. DESIGN: Retrospective case series. SETTING: Tertiary care, university-affiliated, nonprofit teaching hospital. PATIENTS: 3707 patients who had CABG between 1 February 1996 and 1 August 1997. MEASUREMENTS: Chest radiographs were reviewed, and information on pleural fluid findings, pleural effusion treatment, and cardiac surgery was obtained from medical records and a cardiac surgery database. RESULTS: Pleural effusions that occupied more than 25% of the hemithorax were found in 29 patients (0.78%). Seven of the effusions were attributed to congestive heart failure, 2 were attributed to pericarditis, and 1 was attributed to pulmonary embolism. The explanation for the remaining 19 effusions was unclear. All but 2 effusions were predominantly left-sided. Of these 19 effusions, 8 were bloody and 11 were nonbloody. Bloody effusions usually occurred earlier, contained higher lactic acid dehydrogenase levels, and were frequently eosinophilic. Nonbloody effusions tended to be more difficult to manage. CONCLUSIONS: Large pleural effusions may develop in a small proportion of patients after CABG. The cause of many of these effusions is unclear. Most bloody effusions can be managed with one to three therapeutic thoracenteses. Resolution of nonbloody effusions may require anti inflammatory agents, tube thoracostomy, or intrapleural injection of sclerosing agents. PMID- 10375338 TI - The effect of low-dose continuous estrogen and progesterone therapy with calcium and vitamin D on bone in elderly women. A randomized, controlled trial. AB - BACKGROUND: Hormone replacement therapy (HRT), the mainstay of osteoporosis prevention, is limited because of dose-related risks, side effects, and patient acceptance. The bone-sparing efficacy and tolerability of the lowest available doses of HRT have not been adequately studied in elderly women. OBJECTIVE: To determine the bone-sparing effect of continuous low-dose HRT in elderly women. DESIGN: Randomized, double-blind, placebo-controlled trial. SETTING: University osteoporosis research and clinical center. PATIENTS: 128 healthy white women (age > 65 years) with low bone mass recruited by word of mouth and by local advertisement. The principal eligibility criterion was spinal bone mineral density of 0.90 g/cm2 or less. INTERVENTION: Continuous therapy with conjugated equine estrogen, 0.3 mg/d, and medroxyprogesterone, 2.5 mg/d, or matching placebo. Sufficient calcium supplementation was given to bring all calcium intakes above 1000 mg/d in both groups; supplemental oral 25-hydroxyvitamin D was given to maintain serum 25-hydroxyvitamin D levels of at least 75 nmol/L in both groups. MEASUREMENTS: Bone mineral density of the spine, hip, total body, and forearm; serum total alkaline phosphatase and serum osteocalcin levels at 6-month intervals; and 24-hour urine creatinine and hydroxyproline excretion at baseline, 12 months, and 42 months. RESULTS: During 3.5 years of observation, spinal bone mineral density increased by 3.5% (P < 0.001) in an intention-to-treat analysis and by 5.2% among patients with greater than 90% adherence to therapy. Significant increases were seen in total-body and forearm bone density (P < 0.01). Symptoms related to HRT (breast tenderness, spotting, pelvic discomfort, and mood changes) were mild and short-lived. CONCLUSIONS: Continuous low-dose HRT with conjugated equine estrogen and oral medroxyprogesterone combined with adequate calcium and vitamin D provides a bone-sparing effect that is similar or superior to that provided by other, higher-dose HRT regimens in elderly women. This combination is well tolerated by most patients. PMID- 10375339 TI - Treatment of the systemic capillary leak syndrome with terbutaline and theophylline. A case series. AB - BACKGROUND: The systemic capillary leak syndrome is a rare idiopathic disorder characterized by recurrent episodes of hypotension and hemoconcentration due to sudden transient extravasation of 10% to 70% of plasma. Mortality rates 5 years after diagnosis have been reported to be 76%. OBJECTIVE: To assess the efficacy of a prophylactic regimen for the systemic capillary leak syndrome. DESIGN: Case series. SETTING: Tertiary referral center. PATIENTS: Eight patients followed over the past 18 years. INTERVENTION: Oral terbutaline plus aminophylline or theophylline. MEASUREMENTS: Long-term clinical follow-up. RESULTS: During a median follow-up of 9 years (range, 2 to 18 years), two patients (25%) died: one during an acute episode and one of complications related to long-term corticosteroid therapy. The other six patients are alive and healthy. The frequency and severity of the episodes decreased by a median of 30-fold. Recurrences were associated with decreased serum theophylline levels, possibly caused by enzyme induction or autoinduction. The extended-release form of medication was more successful. Sympathomimetic side effects were significant. CONCLUSIONS: A regimen of terbutaline and theophylline seems to be effective prophylaxis against the systemic capillary leak syndrome. Maintenance of therapeutic drug levels was associated with favorable results. PMID- 10375340 TI - Functional somatic syndromes. AB - The term functional somatic syndrome has been applied to several related syndromes characterized more by symptoms, suffering, and disability than by consistently demonstrable tissue abnormality. These syndromes include multiple chemical sensitivity, the sick building syndrome, repetition stress injury, the side effects of silicone breast implants, the Gulf War syndrome, chronic whiplash, the chronic fatigue syndrome, the irritable bowel syndrome, and fibromyalgia. Patients with functional somatic syndromes have explicit and highly elaborated self-diagnoses, and their symptoms are often refractory to reassurance, explanation, and standard treatment of symptoms. They share similar phenomenologies, high rates of co-occurrence, similar epidemiologic characteristics, and higher-than-expected prevalences of psychiatric comorbidity. Although discrete pathophysiologic causes may ultimately be found in some patients with functional somatic syndromes, the suffering of these patients is exacerbated by a self-perpetuating, self-validating cycle in which common, endemic, somatic symptoms are incorrectly attributed to serious abnormality, reinforcing the patient's belief that he or she has a serious disease. Four psychosocial factors propel this cycle of symptom amplification: the belief that one has a serious disease; the expectation that one's condition is likely to worsen; the "sick role," including the effects of litigation and compensation; and the alarming portrayal of the condition as catastrophic and disabling. The climate surrounding functional somatic syndromes includes sensationalized media coverage, profound suspicion of medical expertise and physicians, the mobilization of parties with a vested self-interest in the status of functional somatic syndromes, litigation, and a clinical approach that overemphasizes the biomedical and ignores psychosocial factors. All of these influences exacerbate and perpetuate the somatic distress of patients with functional somatic syndromes, heighten their fears and pessimistic expectations, prolong their disability, and reinforce their sick role. A six-step strategy for helping patients with functional somatic syndromes is presented here. PMID- 10375341 TI - Recent advances in varicella-zoster virus infection. AB - Varicella-zoster virus has developed a complex strategy that allows it to remain latent in the body and avoid destruction by the immune system. Although varicella and zoster have been recognized since antiquity, several new clinical syndromes- including chronic chickenpox with persistent verrucous lesions and disseminated varicella without skin lesions--have been noted in patients with AIDS. Acyclovir has been the mainstay for treating severe varicella-zoster virus infections; however, newer antiviral agents, including valacyclovir and famciclovir, have expanded therapeutic options for treating adults with herpes zoster. The recently licensed live attenuated vaccine for varicella-zoster virus is effective in preventing chickenpox, and the vaccine's ability to stimulate immunity in seropositive adults suggests a promising strategy with which to modify the course of herpes zoster. PMID- 10375342 TI - Evaluating novel cardiovascular risk factors: can we better predict heart attacks? AB - Myocardial infarction often occurs among persons without traditional risk factors, and it has been hypothesized that assessment of "novel" markers may help identify persons who are prone to premature atherothrombosis. However, when considering the clinical utility of screening for any new marker for cardiovascular disease, physicians should consider whether there is a standardized and reproducible assay for the marker of interest; whether there is a consistent series of prospective epidemiologic studies indicating that baseline elevations of the novel marker predict future risk; and whether assessment of the novel marker adds to the predictive value of other plasma-based risk factors, specifically, the ratio of total cholesterol to high-density lipoprotein cholesterol. In this article, these criteria are used to evaluate five promising markers of cardiovascular risk: lipoprotein(a), total plasma homocysteine, fibrinolytic capacity, fibrinogen, and high-sensitivity C-reactive protein. Background is also provided to assist physicians in deciding whether one or more of these novel markers deserve clinical consideration in general outpatient settings. PMID- 10375343 TI - Genes and obesity: is there reason to change our behaviors? PMID- 10375344 TI - The other side of the bed rail. PMID- 10375345 TI - Epinephrine and outcome after cardiac arrest. PMID- 10375346 TI - Epinephrine and outcome after cardiac arrest. PMID- 10375347 TI - Bee sting dysphagia. PMID- 10375348 TI - Bee sting dysphagia. PMID- 10375349 TI - Electronic patient-physician communication. PMID- 10375350 TI - Venlafaxine-associated hepatitis. PMID- 10375351 TI - Toward an understanding of frailty. PMID- 10375352 TI - Function and evolution of Otx proteins. AB - Otx proteins comprise an important class of homeodomain-containing transcription factors known for their essential roles in anterior head formation. Here, we briefly review the basic structural features and functional diversity of Otx proteins and describe current views on the evolution of Otx genes in metazoans. A prominent feature of Otx homeodomains is a lysine residue at position 9 of the recognition helix, which confers high-affinity binding to TAATCC/T elements on DNA. Besides their DNA binding properties, surprisingly little is known about how Otx proteins function to activate target genes in selective regions of the embryo. While an essential and ancient role for Otx is to pattern the anterior regions of the head, drawing conclusions about primordial functions is difficult. This is because Otx proteins have been recruited for numerous developmental roles, and derived functions have often evolved to meet the specialized requirements of individual taxonomic groups. In sea urchin embryos, one form of Otx may have been co-opted by the Wnt--catenin signaling pathway. The consequence of such an evolutionary event would be to link a highly conserved signal transduction pathway to a set of novel downstream genes that make use of Otx for their transcription. PMID- 10375353 TI - Recent advances in neonatology. PMID- 10375354 TI - Longitudinal study of behaviour disorders in low birthweight infants. AB - AIM: To compare the prevalence of childhood and adolescent behavioural problems in low birthweight infants with matched controls. METHODS: A cohort study of a geographically defined population of survivors of /=2 years, the differences in neurodevelopmental outcome among the two groups were not significant. CONCLUSIONS: In neonates treated with surfactant moderate and severe pulmonary haemorrhage is associated with an increased risk of death and short term morbidity. Pulmonary haemorrhage does not seem to be associated with increased long term morbidity. PMID- 10375362 TI - Iron nutritional status in preterm infants fed formulas fortified with iron. AB - AIMS: To prospectively evaluate the iron nutritional status of preterm infants fed either a term (0.5 mg/dl iron) or preterm (0.9 mg/dl) formulas fortified with iron after hospital discharge. METHODS: Healthy low birthweight preterm infants were randomly assigned into three groups at the time of hospital discharge. Group A were fed an iron fortified preterm formula (0.9 mg/dl iron) until 6 months corrected age; group B, a fortified term formula (0.5 mg/l iron) until 6 months corrected age group C, the preterm formula between hospital discharge and term, then the term formula until 6 months corrected age. RESULTS: Seventy eight infants were followed up to 6 months corrected age. Iron intake from formula differed significantly between the groups (A, 1.17 mg/kg/day (SD 0.32) > C, 0. 86 mg/kg/day (SD 0.40) = B, 0.81 mg/kg/day (SD 0.23); p < 0.0001). Haemoglobin concentrations were similar to those of iron sufficient preterm infants of the same postnatal age, and term infants of the same postmenstrual age (after 3 months of age). There were no significant differences in haemoglobin concentration (p = 0.391), plasma ferritin (A vs B, p = 0.322), or in the incidence of iron deficiency (A vs B, p = 0.534). CONCLUSIONS: Iron fortified formulas containing between 0.5 and 0.9 mg/dl iron seem to meet the iron nutritional needs of preterm infants after hospital discharge. PMID- 10375363 TI - Knee-heel length measurement in healthy preterm infants. AB - AIM: To examine the reproducibility of crown-heel length measurement; the precision and reproducibility of knee-heel length measurement; and the association between the two in healthy preterm infants. METHODS: Paired crown heel and knee-heel lengths were measured on 172 occasions by three observers in 43 preterm infants between 205 and 458 days of postconceptional age. RESULTS: Crown-heel length (CHL) measurement was highly reproducible, with a coefficient of variation (CV) of 0.41%. Knee-heel length (KHL) measurement was relatively precise (CV 0.78%), but less reproducible (intra-observer CV 1.77%, intra observer CV 2.11%), especially in larger infants. The association between KHL and CHL was not consistent and varied with age. KHL was a poor predictor of CHL, with a 95% predictive interval of +/- 27.5 mm. CONCLUSIONS: KHL was less reproducible than CHL, especially in larger infants, and a poor predictor of CHL. PMID- 10375364 TI - Neonatal bilirubin production, reflected by carboxyhaemoglobin concentrations, in Down's syndrome. AB - AIM: To determine whether increased bilirubin production, reflected by blood carboxyhaemoglobin (COHb) values, is responsible for hyperbilirubinaemia in cases of Down's syndrome with no obvious cause for excessive jaundice. METHODS: Blood was sampled on the third day of life for COHb, total haemoglobin (tHb), and serum total bilirubin, from 19 consecutively born neonates with Down's syndrome (a subset of 34 term babies), who had developed hyperbilirubinaemia (serum bilirubin >/= 256 micromol), and from 32 term controls. COHb, measured by gas chromatography, was corrected for inspired CO (COHbc) and expressed as a percentage of tHb. RESULTS: Significantly more of the Down's syndrome subset developed hyperbilirubinaemia than the controls (10/19 (52%) vs 7/32 (22%), relative risk 2.4, 95% confidence intervals (CI) 1.10 to 5.26). Third day serum bilirubin values (mean (SD)) were higher in the Down's syndrome neonates than in controls (214 +- 63 micromol/l vs 172 +- 54 micromol/l, respectively, p=0.015). Mean (SD) COHbc values were significantly higher in the Down's syndrome neonates than in controls (0.92 +- 0. 24% vs 0.63 +- 0.17%; p<0.0001). However, Down's syndrome neonates who became hyperbilirubinaemic had similar COHbc values to those who did not (0.87 +- 0.26% and 0.95 +- 0.23%, respectively). These values contrast with those of the controls, in whom a significant increase in COHbc was associated with hyperbilirubinaemia (0.74 +- 0. 15% vs 0.60 +- 0.16%, respectively; p<0.05). tHb values were similar in both groups. CONCLUSIONS: Down's syndrome neonates had a greater risk of hyperbilirubinaemia, and higher COHbc values, than controls. However, excessive bilirubin production could not be exclusively responsible for the hyperbilirubinaemia. By inference, decreased bilirubin elimination probably plays a greater part in its pathogenesis than in controls. Down's syndrome neonates may have abnormal erythropoiesis, leading to increased haem turnover. PMID- 10375365 TI - Incidence and risk factors for ventricular septal defect in "low risk" neonates. AB - AIMS: To quantify the incidence of ventricular septal defect in "low-risk" neonates; and to define any associated risk factors. METHODS: One hundred and seventy three patients with ventricular septal defects from a scanned population of 3971 clinically normal neonates were compared with scanned controls, considered to be clinically normal. A subset of the group with defects was compared with normal infants delivered over the same period, to identify any seasonal variation. RESULTS: Ventricular septal defects were detected in 4.36 % of the "scanned" group (173 out of 3971). Ten had perimembranous defects and the remainder apical or muscular lesions. Eleven neonates had multiple defects. The incidence of ventricular septal defect was independent of most tested risk factors. There were significantly more girls in the group with defects compared with the controls (p = 0.004). The defects group also contained fewer infants born during summer months (p = 0.04). CONCLUSIONS: The incidence of ventricular septal defects was much higher than might be expected, given that "high risk infants" were excluded. The observations that gender and season of birth affect the rate suggest that both genetic and environmental factors may be involved in the aetiology. PMID- 10375366 TI - Safety and effectiveness of BCG vaccination in preterm babies. AB - AIM: To assess the cell mediated immune response to BCG vaccine in preterm babies. METHODS: Sixty two consecutive preterm babies born at < 35 weeks of gestation were randomly allocated into two groups. Babies in group A were vaccinated early at 34-35 weeks and group B were vaccinated late at 38-40 weeks of postconceptional age. The two groups were similar in terms of: gestational age (mean (SD) 33.1 (1. 1) and 33 (1.2) weeks, respectively); birthweight 1583 (204) and 1546 (218) g; neonatal problems; socioeconomic status; and postnatal weight gain. The cell mediated immune response to BCG was assessed using the Mantoux test and the lymphocyte migration inhibition test (LMIT) 6-8 weeks after BCG vaccination. Induration of >5 mm after the Mantoux test was taken as a positive response. RESULTS: There was no significant difference in the tuberculin conversion rates (80% and 80.7%, respectively), positive LMIT (86.6% and 90.3%, respectively), or BCG scar (90.0% and 87.1%, respectively) among the two groups. CONCLUSIONS: Prematurity seems to be an unlikely cause for poor vaccine uptake. Preterm babies can be effectively vaccinated with BCG at 34-35 weeks of postconceptional age, the normal time of discharge in a developing country. PMID- 10375367 TI - Severe apnoeas following immunisation in premature infants. AB - Four premature infants developed apnoeas severe enough to warrant resuscitation after immunisation with diphtheria, pertussis, and tetanus (DPT), and Haemophilus influenzae B (Hib). One required re-intubation and ventilation. Although apnoeas after immunisation are recognised, they are not well documented. It is time for further research to elucidate the best time to immunise such infants. PMID- 10375368 TI - Neonatal varicella: varicella zoster immunoglobulin (VZIG) does not prevent disease. AB - Two infants with severe varicella are reported. They received varicella zoster immunoglobulin (VZIG) without concurrent information to parents or carers regarding further care. In both these cases there was a three day delay between the onset of symptoms and initiation of aciclovir. This delay was due to lack of awareness of the high risk of varicella in these infants. Infants born to mothers with onset of chickenpox 4 days before to 2 days after delivery are at risk of fatal varicella, despite the use of VZIG prophylaxis. PMID- 10375369 TI - Low birthweight and adult insulin resistance: the "catch-up growth" hypothesis. PMID- 10375370 TI - Bartholomew Mosse (1712-59), Sir Fielding Ould (1710-89), and the Rotunda Hospital, Dublin. PMID- 10375371 TI - Detection of heart disease in infancy. PMID- 10375372 TI - Microdomain arrangement of the SERCA-type Ca2+ pump (Ca2+-ATPase) in subplasmalemmal calcium stores of paramecium cells. AB - We localized SERCA pumps to the inner region of alveolar sac membranes, facing the cell interior, by combining ultrastructural and biochemical methods. Immunogold labeling largely predominated in the inner alveolar sac region which displayed aggregates of intramembrane particles (IMPs). On image analysis, these represented oligomeric arrangements of approximately 8-nm large IMP subunits, suggesting formation of SERCA aggregates (as known from sarcoplasmic reticulum). We found not only monomers of typical molecular size ( approximately 106 kD) but also oligomeric forms on Western blots (using anti-SERCA antibodies, also against endogenous SERCA from alveolar sacs) and on electrophoresis gelautoradiographs of 32P-labeled phosphoenzyme intermediates. Selective enrichment of SERCA-pump molecules in the inner alveolar sac membrane region may eliminate Ca2+ after centripetal spread observed during exocytosis activation, while the plasmalemmal Ca2+ pump may maintain or reestablish [Ca2+] in the narrow subplasmalemmal space between the outer alveolar sac membrane region and the cell membrane. We show for the first time the microzonal arrangement of SERCA molecules in a Ca2+ store of a secretory system, an intensely discussed issue in stimulus-secretion coupling research. PMID- 10375373 TI - Immunohistochemical localization of EphA5 in the adult human central nervous system. AB - To better understand the functional role of EphA5 in the adult human central nervous system (CNS), we performed an immunohistochemical mapping study. EphA5, like other members of the Elk/Eph family of receptor tyrosine kinases, was widely distributed in CNS neurons. However, the distribution of the neuronal staining was not uniform. The abundance of stained neurons appeared to increase from the forebrain to the hindbrain and spinal cord. Glial and endothelial tissue was unstained. These findings are consistent with the existence of receptor and ligand gradients in different brain regions. The localization of EphA5 to motor and sensory neurons is consistent with a role of EphA5 in neural plasticity, cell cell recognition, and topographical orientation of neuronal systems. PMID- 10375375 TI - In vivo antibody-mediated modulation of aminopeptidase A in mouse proximal tubular epithelial cells. AB - Aminopeptidase A (APA) is one of the many renal hydrolases. In mouse kidney, APA is predominantly expressed on the brush borders and sparsely on the basolateral membranes of proximal tubular epithelial cells. However, when large amounts of monoclonal antibodies (MAbs) against APA were injected into mice, we observed strong binding of the MAbs to the basolateral membranes, whereas the MAbs bound only transiently to the brush borders of the proximal tubular epithelial cells. In parallel, APA itself disappeared from the brush borders by both endocytosis and shedding, whereas it was increasingly expressed on the basolateral sides. Using ultrastructural immunohistology, we found no evidence for transcellular transport of endocytosed APA to the basolateral side of the proximal tubular epithelial cells. The absence of transcellular transport was confirmed by experiments in which we used a low dose of the MAbs. Such a low dose did not result in binding of the MAbs to the brush borders and had no effect on the presence of APA in the brush borders of the proximal tubular epithelial cells. In these experiments we still could observe binding of the MAbs to the basolateral membranes in parallel with the local appearance of APA. In addition, treatment of mice with chlorpromazine, a calmodulin antagonist that interferes with cytoskeletal function, largely inhibited the MAb-induced modulation of APA. Our studies suggest that injection of MAbs to APA specifically interrupts the normal intracellular traffic of this enzyme in proximal tubular epithelial cells. This intracellular transport is dependent on the action of cytoskeletal proteins. PMID- 10375374 TI - Expression of REG protein during cell growth and differentiation of two human colon carcinoma cell lines. AB - We localized REG protein in Paneth cells and nonmature columnar cells of the human small intestinal crypts and speculated that this protein was associated with growth and/or differentiation. The aim of this study was to determine whether REG protein is present in two human colon cancer cell lines that exhibit enterocytic differentiation after confluence and to investigate changes in the level of its expression during growth and differentiation. Results were compared to those obtained on cells that remain undifferentiated. Western immunoblotting and immunofluorescence demonstrated the presence of REG protein in the three cell lines. With the antisera against human REG protein, the staining was diffusely spread throughout the cytoplasm at Day 2, and after Days 3-4 it appeared to have migrated to cell boundaries. After confluence, we observed only a punctate staining array along cell boundaries, which disappeared at Day 15. REG mRNA expression was demonstrated by RTPCR and REG mRNA hybridization until Day 13, but not after, in the three cell types. REG protein may be involved in cellular junctions. Its presence appears to be associated with the cell growth period and the protein must be downregulated when growth is achieved and differentiation is induced. PMID- 10375376 TI - Sulfated glycosaminoglycans in guinea pig neutrophils studied by use of cationic colloidal gold. AB - Using a high electron resolution staining method, cationic colloidal gold (CCG, pH 1.0) staining, we studied the fine structural localization of sulfated glycosaminoglycans (GAGs) in various maturational stages of guinea pig neutrophils. Azurophil and specific granules of neutrophils reacted positively to CCG, with variety in labeling according to maturation. All immature azurophil and specific granules were labeled selectively. Mature granules lost their affinity with CCG. CCG-positive labeling was also observed in the trans to trans-most Golgi apparatus of promyelocytes and myelocytes. Prior absorption with poly-l lysine prevented CCG labeling of tissue sections. Mild methylation of ultrathin sections at 37C did not alter CCG labeling, whereas CCG labeling disappeared after active methylation at 60C. Treatment with chondroitinase ABC or heparinase I abolished the majority of CCG labeling. These findings suggest the existence of sulfated GAGs not only in immature azurophil but also in immature specific granules of neutrophils. Sulfation of GAGs occurs in the trans- to trans-most Golgi apparatus of neutrophil granulocytes. A possible correlation between accumulation of sulfated GAGs and maturation of specific granules in neutrophils is also discussed. PMID- 10375377 TI - Presence of H type 3/4 chains of ABO histo-blood group system in serous cells of human submandibular gland and regulation of their expression by the secretor gene (FUT2). AB - We have investigated by immunochemistry the distribution of H Type 3/4 chains of the ABO histo-blood group system in human submandibular gland using a monoclonal anti-H MBr1 antibody specific for H Type 3/4 chains, and have found the expression of H Type 3/4 chains was mainly in the serous cells. Serous cells from secretors were stained by MBr1 but not by anti-A and anti-B antibodies, whereas serous cells from nonsecretors exhibited a negative reaction with MBr1. Mucous cells were not stained by MBr1. Only a few striated duct cells showed a weak reaction with anti-H MBr1. These results suggested that the H Type 3/4 chains were distributed predominantly in the serous cells of the human submandibular gland and that secretor Type alpha(1,2)fucosyltransferase (Se enzyme) controlled the synthesis of H Type 3/4 chains in vivo. Saliva also contained H Type 3/4 chains, which were controlled by the secretor gene (FUT2). The differences in the distributions of H Type 1, H Type 2, and H Type 3/4 chains of the ABO histo blood group system in the submandibular gland are discussed. PMID- 10375378 TI - Differential expression of the cyclic GMP-stimulated phosphodiesterase PDE2A in human venous and capillary endothelial cells. AB - We developed selective monoclonal antibodies and used them for Western and immunocytochemical analyses to determine the tissue and cellular distribution of the human cyclic GMP-stimulated phosphodiesterase (PDE2). Western analysis revealed PDE2A expression in a variety of tissue types, including cerebellum, neocortex, heart, kidney, lung, pulmonary artery, and skeletal muscle. Immunocytochemical analysis revealed PDE2A expression in a subset of tissue endothelial cells. PDE2A immunostaining was detected in venous and capillary endothelial cells in cardiac and renal tissue but not in arterial endothelial cells. These results were confirmed by in situ hybridization. PDE2A immunostaining was also absent from luminal endothelial cells of large vessels, such as aorta, pulmonary, and renal arteries, but was present in the endothelial cells of the vasa vasorum. PDE2A immunostaining was detected in the endothelial cells of a variety of microvessels, including those in renal and cardiac interstitial spaces, renal glomerulus, skin, brain, and liver. Although PDE2A was not readily detected in arterial endothelial cells by immunocytochemistry of intact tissue, it was detected at low levels in cultured arterial endothelial cells. These results suggest a possible role for PDE2A in modulating the effects of cyclic nucleotides on fluid and inflammatory cell transit through the endothelial cell barrier. PMID- 10375379 TI - Connexin45, a major connexin of the rabbit sinoatrial node, is co-expressed with connexin43 in a restricted zone at the nodal-crista terminalis border. AB - The pacemaker of the heart, the sinoatrial (SA) node, is characterized by unique electrical coupling properties. To investigate the contribution of gap junction organization and composition to these properties, the spatial pattern of expression of three gap junctional proteins, connexin45 (Cx45), connexin40 (Cx40), and connexin43 (Cx43), was investigated by immunocytochemistry combined with confocal microscopy. The SA nodal regions of rabbits were dissected and rapidly frozen. Serial cryosections were double labeled for Cx45 and Cx43 and for Cx40 and Cx43, using pairs of antibody probes raised in different species. Dual channel scanning confocal microscopy was applied to allow simultaneous visualization of the different connexins. Cx45 and Cx40, but not Cx43, were expressed in the central SA node. The major part of the SA nodal-crista terminalis border revealed a sharply demarcated boundary between Cx43-expressing myocytes of the crista terminalis and Cx45/Cx40-expressing myocytes of the node. On the endocardial side, however, a transitional zone between the crista terminalis and the periphery of the node was detected in which Cx43 and Cx45 expression merged. These distinct patterns of connexin compartmentation and merger identified suggest a morphological basis for minimization of contact between the tissues, thereby restricting the hyperpolarizing influence of the atrial muscle on the SA node while maintaining a communication route for directed exit of the impulse into the crista terminalis. PMID- 10375380 TI - Multistratified expression of polysialic acid and its relationship to VAChT containing neurons in the inner plexiform layer of adult rat retina. AB - We investigated the localization of polysialic acid (PSA), neural cell adhesion molecule (NCAM), and vesicular acetylcholine transporter (VAChT) in adult rat retina by using immunofluorescence with a confocal laser scanning microscope. Western blot analysis showed a typical broad smear of PSA and isoforms of NCAM (120, 140, and 180 kD). PSA immunofluorescence revealed multistratification in the inner plexiform layer (IPL). Dual immunostaining for PSA and NCAM exhibited the selective co-expression of PSA and NCAM on Muller cells. Moreover, dual immunolabeling for PSA and VAChT completely separated the five strata in the IPL. Strata 1, 3, and 5 were immunoreactive for PSA and Strata 2 and 4 for VAChT. These results suggest the possibility that PSA molecules on Muller cells are spatially related to ON and OFF retinal channels in the IPL. PMID- 10375381 TI - Immunocytochemical localization of phospholipase C isozymes in cord blood and adult T-lymphocytes. AB - The response of T-cells to peptide antigen plus major histocompatibility complex (MHC) consists of a series of cellular events collectively called T-cell activation. An essential component of this pathway is phospholipase C (PLC)gamma1, whose hydrolytic activity increases rapidly after binding of ligands to the T-cell receptor (TCR) and consequent activation of tyrosine kinases. Recent studies also suggest a GTP binding protein-dependent activation of PLCbeta during the early steps of T-cell activation. On the basis of these findings, we first checked the expression of PLC isoforms by Western blotting and by confocal and electron microscopy techniques, and then we looked for the phosphoinositide breakdown induced by CD3 engagement in cord and adult T-lymphocytes. Our results indicated that PLCbeta1 was almost exclusively expressed in cord T-cells, whereas PLCbeta2 was more strongly represented in the adult. The amount of PLCgamma1 was found to be larger in the adult than in cord cells. No significant differences were found in PLCgamma2 and delta2 expression. PLCdelta1 was scarcely detectable. On CD3 stimulation, adult lymphocytes gave rise, as expected, to a dramatic increase in phosphoinositide breakdown, whereas in cord cells this response was scarcely detected. These results indicate that a shift in PLC expression occurs in the postnatal period and that this change is associated with induction of the capability to respond to CD3 engagement with phosphoinositide hydrolysis. PMID- 10375382 TI - Differential expression of gap junction proteins connexin26, 32, and 43 in normal and crush-injured rat sciatic nerves. Close relationship between connexin43 and occludin in the perineurium. AB - We immunohistochemically and morphometrically examined the expression of gap junction protein connexin (Cx) in normal and crush-injured rat sciatic nerves using confocal laser scanning microscopy. Cx26 was localized in the perineurium and Cx43 was present in the perineurium and the epineurium, whereas Cx32 was confined to the paranodal regions of the nodes of Ranvier. Double labeling for connexins and laminin revealed that Cx43 was localized in multiple layers of the perineurium, whereas Cx26 was confined to the innermost layer. Double labeling for connexins and a tight junction protein, occludin, showed that occludin frequently coexisted with Cx43 but existed separately from Cx26 in the perineurium. After crush injury, the pattern of normal Cx32 expression was initially lost but recovered, whereas Cx43 rapidly appeared in the endoneurium and its expression was subsequently attenuated. Although crush injury produced no apparent alteration in Cx43 and occludin in the perineurium, a rapid increase and a subsequent decrease in the frequency of Cx26-positive spots during nerve regeneration were shown by morphometric analysis. These results indicate that Cx26, Cx32, and Cx43 are expressed differently in various types of cells in peripheral nerves and that their expressions are differentially regulated after injury. The expression of connexins and occludin in the perineurium suggests that perineurial cells are not uniform in type and that the regulation of gap junctions and tight junctions is closely related in the perineurium. PMID- 10375383 TI - Specificity of the localization of transforming growth factor-alpha immunoreactivity in colon mucosa. AB - Transforming growth factor-alpha (TGF-alpha) plays an important role in gastrointestinal pathophysiology. However, the exact location of its expression in the intestine is still controversial. This study systematically compared the localization of TGF-alpha immunoreactivity in frozen or fixed human colon using three different antibodies and examined specificity of antibodies by using tissues from TGF-alpha knockout mice and by Western blotting. Consistent with the mRNA distribution revealed by in situ hybridization, a similar staining pattern was obtained in frozen sections by all three antibodies, localizing on the surface and along the crypt epithelium. In paraffin sections, although the polyclonal antibodies (raised against recombinant human or rat TGF-alpha) gave minimal staining, the monoclonal antibody (against C-terminal peptide of human TGF-alpha) still gave intense staining on the surface and upper crypt epithelium. By using specimens from TGF-alpha knockout mice in immunostaining and Western blotting, the polyclonal antibodies were shown to be specific. In contrast, specificity of the monoclonal antibody was in doubt in rodent tissues because it gave similar detection between wild-type and knockout mice in both analyses, indicating its crossreaction to non-TGF-alpha molecules. In conclusion, frozen sections and antibodies raised from recombinant TGF-alpha should be used for TGF alpha immunohistochemistry in the colon. PMID- 10375384 TI - Immunoglobulin and enzyme-conjugated dextran polymers enhance u-PAR staining intensity of carcinoma cells in peripheral blood smears. AB - The presence of disseminated carcinoma cells in bone marrow and peripheral blood has prognostic importance in patients with carcinomas. Much evidence indicates that dissemination of tumor cells may depend on activation of a variety of degradative enzymes. A strong positive correlation has been shown between the expression of tumor cell proteases and tumor invasion. Therefore, phenotypic characterization of disseminated carcinoma cells for expression of protease activators might define the invasive potential of the cells. We present an immunocytochemically enhanced staining method that allows phenotyping of disseminated carcinoma cells in bone marrow and peripheral blood smears. In the first step, the cells were incubated with antibodies against urokinase plasminogen activator receptor (u-PAR) and subsequently with secondary antibodies conjugated to peroxidase-labeled dextran polymers. A brown color reaction was developed with diaminobenzidine as chromogen. In the second step, the cells were incubated with alkaline phosphatase-conjugated murine monoclonal antibodies against a common cytokeratin epitope and a red color reaction was developed with new fuchsin as substrate. This method allows simultaneous and unambiguous immunolabeling of intracellular cytokeratin and of u-PAR intracellularly and on the surface of carcinoma cells. This novel approach can be used for detection and phenotyping of carcinoma cells in blood smears for u-PAR or, presumably, for any other heterogeneously expressed antigen on the surface of the detected cells. PMID- 10375385 TI - Enhanced in situ detection of beta-glucuronidase activity in murine tissue. AB - We outline here a protocol for high-resolution in situ localization of beta glucuronidase in murine tissues processed in glycol methacrylate (GMA). Murine tissues were first stained with 5-bromo-4-chloro-3-indolyl-beta-d-glucuronic acid (x-gluc), followed by histological processing in GMA. Retention of the blue indigo reaction product after overnight incubations in x-gluc allowed high resolution localization of beta-glucuronidase activity by brightfield microscopy. When illuminated under darkfield, the x-gluc signal was enhanced, permitting detection even in cells with low-level enzyme activity. This technique offers for the first time a more sensitive enzyme histochemical method of detecting beta glucuronidase activity in animal tissues and also the opportunity to examine expression at high magni-fication. PMID- 10375386 TI - Detection of pathological zinc accumulation in neurons: methods for autopsy, biopsy, and cultured tissue. AB - It has been repeatedly shown that synaptically released zinc contributes to excitotoxic neuronal injury in ischemia, epilepsy, and mechanical head trauma. Such zinc-induced injury leaves an unmistakable "footprint" in the injured neurons, allowing an easy and unambiguous postmortem diagnosis. This footprint is the presence of weakly bound, histochemically reactive zinc in the cytoplasm of the perikaryon and proximal dendrites. Such staining appears to be a necessary and sufficient marker for zinc-induced neuronal injury. Here we show how to prepare and stain tissue from biopsy, autopsy, or experimental animal sources for maximal contrast and visibility of zinc-injured neurons. PMID- 10375387 TI - Heparin induced thrombocytopenia: diagnosis and contemporary antithrombin management. AB - Heparin-induced thrombocytopenia (HIT) may be complicated by severe thrombotic complications and death. Currently no specific laboratory test is available to make the diagnosis. When HIT is clinically suspected, heparin should be discontinued immediately. While no specific therapy for HIT exists, there is increasing evidence that acute antithrombin therapy may significantly reduce morbidity and mortality. Among several agents, the direct antithrombins, such as r-hirudin and argatroban, look the most promising for acute treatment. PMID- 10375388 TI - Efficacy of abciximab induced platelet blockade using a rapid point of care assay. AB - Anciximab provides potent, but variable degrees of platelet inhibition both during the duration of intravenous administration and at 12 hours following therapy. Platelet function was assessed using the PC-RPFA system in 78 patients scheduled for percutaneous coronary revascularization who were administered the standard abciximab weight-adjusted bolus and 12-hour infusion. The PC-RPFA system is a cartridge-based, semiautomated point-of-care whole-blood assay that incorporates fibrinogen-coated polystyrene beads, buffers, and a modified thrombin receptor activating peptide (Isotrap) in lyophilized form. The instrument detects the agglutination rate between the stimulated platelets and the fibrinogen-coated beads, and provides a quantitative digital display in less than 2 minutes. No differences in the level of platelet inhibition were observed in these abciximab-treated patients by diabetic status, gender, smoking, diagnosis (unstable angina, chronic stable angina, recent myocardial infarction), or abciximab treatment status (first time vs. retreatment). Nocorrelation of the PC-RPFA rate of platelet aggregation with clinical demographic factors was observed, with the exception of baseline hematocrit (r2 = 0.4556). The relationship between the PC-RPFA rate of aggregation and hematocrit reflects light absorbance by erythrocytes and is specific to the PC-RPFA system. The absolute rate of platelet aggregation (slope) reported by the PC-RPFA is correlated with percent aggregation, thus making it potentially possible to predict the level of aggregation without reference to a baseline (pretreatment) measure of platelet function. This correlation was closest for patients having <40% baseline aggregation (r2 = 0.55). Thus, PC-RPFA provides a rapid point-of care assessment of platelet function that could allow for adjustment of abciximab dosing to achieve targeted levels of platelet inhibition. The utility of this device to optimize therapy with platelet glycoprotein IIb/IIIa inhibitors is currently being evaluated. PMID- 10375389 TI - Plasmin activation system in restenosis: role in pathogenesis and clinical prediction? AB - During recent years it has become increasingly recognized that the plasmin activation system is involved in the development of atherosclerosis and restenosis. Responsible pathophysiologic mechanisms, however, remain elusive. This review focuses primarily on the clinicians, point of view, suggesting that increases in plasminogen activator inhibitor type-1 (PAI-1) plasma levels after balloon angioplasty or permanently elevated lipoprotein (a) (Lp(a)) plasma levels might be helpful in the prediction of restenosis after coronary angioplasty. In contrast, tissue-type plasminogen activator (tPA) plasma levels appear unrelated to restenosis, and data regarding a possible role of urokinase-type plasminogen activator (uPA) in circulation are not available at present. Furthermore, a new hypothesis on the pathophysiological role of local PAI-1 overexpression as a beneficial negative feedback mechanism to limit excess cellular proliferation in atherogenesis and restenosis is presented. PMID- 10375390 TI - Glycoprotein IIb/IIIa receptor inhibition in interventional cardiology. PMID- 10375391 TI - Antithrombotic effects of BCH 2763, a new direct thrombin inhibitor, in a canine model of venous thrombosis. AB - Deep venous thrombosis (DVT) is a common cardiovascular disease, resulting in significant mortality each year in the United States. Direct thrombin inhibitors represent a new class of drugs that could potentially be better than conventional antithrombotic therapy based on indirect inhibition of coagulation factors with heparin and warfarin. BCH 2763 is a potent, selective bifunctional thrombin inhibitor that blocks both the active catalytic site and the anion binding exosite. The objective of this study is to test the antithrombotic efficacy of BCH 2763 in a canine model of DVT induced through electrolytic injury to the femoral vein. BCH 2763 was administered at three dose levels: 0.125 mg/kg bolus followed by 10 microg/kg/min IV infusion (low-dose; n = 5), 0.25 mg/kg bolus followed by 20 microg/kg/min infusion (mid-dose; n = 5), and 0.75 mg/kg bolus followed by 60 microg/kg/min (high-dose; n = 5). The control group (n = 5) received a 5-ml intravenous bolus of saline followed by a 1 mL/kg/h infusion. The parameters evaluated were changes in activated partial thromboplastin time (aPTT), thrombin time (TT), prothrombin time (PT), time to formation of an occlusive thrombus in the femoral vein, and the amount of venous blood flow delivered over the course of the experiment. There were significant dose dependent increases in aPTT, TT, and PT in the BCH 2763-treated animals compared with the control group. The time to formation of an occlusive thrombus in the control group averaged 69.6 +/- 9 minutes. Treatment with BCH 2763 prolonged the time to occlusion to 126.4 +/- 13 minutes in the low-dose group, 155.4 +/- 17 minutes in the mid-dose group, and 229 +/- 7 minutes in the high-dose group (80% remained patent for the duration of the study), which were all significantly greater than the controls. Femoral venous blood flow was significantly greater in the mid-dose (51 +/- 8%) and the high-dose (70 +/- 6%) groups compared with the control vessels (22 +/- 3%). In conclusion, the results of this study indicate that BCH 2763 is an effective intravenous antithrombotic agent in the canine electrolytic injury model of venous thrombosis. PMID- 10375392 TI - Oxidative modification of apolipoprotein E in human very-low-density lipoprotein and its inhibition by glycosaminoglycans. AB - The mechanism of metal ion-catalyzed oxidative modification of apolipoprotein E (apoE) in human very-low-density lipoprotein (VLDL) and its inhibition by glycosaminoglycan (GAG) was investigated in vitro. The VLDL oxidation catalyzed by Cu2+ led to the lipid peroxidation, the formation of aggregates, and covalent modification of apoE. The modified apoE lost heparin-binding activity. These results suggest that the lipid peroxidation of VLDL and modification of apoE cause impairment of lipid uptake by cells and deposit the oxidized lipids in the tissues. The lipid peroxidation and oxidative modification of apoE in VLDL mediated by Cu2+ and an aqueous radical generator were suppressed by GAG, heparan sulfate, heparin, and chondroitin sulfate A, even though GAGs demonstrated no ability to scavenge alpha,alpha-diphenyl-beta-picrylhydrazyl radical. There were no relationships between inhibitory activity of GAGs in the VLDL oxidation and their number of sulfate groups which possess chelating activity of metal ion. Therefore, it can be considered that the inhibition of VLDL oxidation by GAGs is possibly due to the interaction between GAG and VLDL which bring about the steric hindrance, interference with the reaction between VLDL particle and the reactive oxygen species. These studies suggest that GAGs preserve the biological functions of apoE from oxidative stress. PMID- 10375394 TI - The subcellular distribution of protein kinase Calpha, -epsilon, and -zeta isoforms during cardiac cell differentiation. AB - There is little information on the molecular events that control the subcellular distribution of protein kinase C during cardiac cell differentiation. We examined protein kinase C activity and the subcellular distribution of representatives of the "classical," "novel," and "atypical" protein kinase C's in P19 murine teratoma cells induced to undergo differentiation into cardiac myocytes by the addition of dimethylsulfoxide to the medium (Grepin et al., Development 124, 2387 2395, 1997). Differentiation was assessed by the presence of striated myosin, a morphological marker for cardiac cells. Addition of dimethyl sulfoxide to the medium resulted in the appearance of striated myosin by 10 days postincubation. Immunolocalization and Western blot studies revealed that a significant proportion of protein kinase Calpha, -epsilon, and -zeta were associated with the particulate fraction in P19 cells prior to differentiation. Differentiation into cardiac cells resulted in a translocation of protein kinase C activity from the particulate fraction to cytosol and localization of most of protein kinase Calpha, -epsilon, and -zeta to the cytoplasmic compartment. The total cellular protein kinase C activity was unaltered during differentiation. The translocation of protein kinase C activity during differentiation of P19 cells into cardiac myocytes was associated with a decrease in the levels of cellular 1, 2-diacyl-sn glycerol. The cellular levels of phosphatidylserine and phosphatidylinositol did not change during differentiation. Addition of 1,2-dioctanoyl-sn-glycerol, a cell permeant 1, 2-diacyl-sn-glycerol analog, reversed the differentiation-induced switch in the relative distribution of protein kinase C activity and dramatically increased the association of protein kinase Calpha with the particulate fraction. Addition of 1,2-dioctanoyl-sn-glycerol did not reverse the pattern of distribution for protein kinase Cepsilon or -zeta. The results indicate that protein kinase C activity and protein kinase Calpha, -epsilon and -zeta isoforms are redistributed from the particulate to the cytosolic fraction during differentiation of P19 cells into cardiomyocytes. The mechanism for the redistribution of protein kinase Calpha may be related to the reduction in the cellular 1,2-diacyl-sn-glycerol levels that accompany differentiation. PMID- 10375393 TI - S-Adenosyl-L-methionine:salicylic acid carboxyl methyltransferase, an enzyme involved in floral scent production and plant defense, represents a new class of plant methyltransferases. AB - S-Adenosyl-L-methionine:salicylic acid carboxyl methyltransferase (SAMT) was partially purified from petals of the annual California plant Clarkia breweri. SAMT catalyzes the formation of methylsalicylate, an important floral scent compound in C. breweri, from salicylic acid and S-adenosyl-L-methionine (SAM). The native enzyme is a dimer with a subunit molecular weight of 40.3 kDa, and it has a Km for salicylic acid of 24 microM and a Km for SAM of 9 microM. A cDNA encoding SAMT was isolated from a C. breweri cDNA library prepared from floral mRNA. The sequence of the protein encoded by SAMT cDNA shows no significant sequence similarity to any protein in the data bank whose biochemical function is known. It does show significant sequence similarity (20-40% identity) to proteins encoded by at least seven Arabidopsis thaliana genes whose sequences have recently been determined in large-scale sequencing projects. The C. breweri SAMT cDNA was expressed in E. coli and the bacterial cells synthesized a functional SAMT protein with properties nearly identical to those of the plant-purified enzyme. PMID- 10375395 TI - A novel phospholipase A2, BJ-PLA2, from the venom of the snake Bothrops jararaca: purification, primary structure analysis, and its characterization as a platelet aggregation-inhibiting factor. AB - This paper describes the isolation and primary structure analysis of a new phospholipase A2 with platelet-aggregation-inhibiting activity from the venom of Bothrops jararaca. The protein, named BJ-PLA2, was isolated by means of ammonium sulfate precipitation and anion-exchange and reversed-phase chromatographies and behaved as a homogeneous single-chain protein on SDS-PAGE. Its amino acid sequence was determined by N-terminal sequencing and analysis of overlapped chemical and proteolytic fragments by automated Edman degradation and mass spectometry determination. BJ-PLA2 consists of 124 amino acid residues and has the structural features of snake venom class II phospholipases A2. Chemical modification with p-bromophenacylbromide caused complete loss of enzymatic activity and partially affected the platelet-aggregation-inhibiting activity of BJ-PLA2. PMID- 10375396 TI - The 650-kDa 12(S)-hydroxyeicosatetraenoic acid binding complex: occurrence in human platelets, identification of hsp90 as a constituent, and binding properties of its 50-kDa subunit. AB - A cytosolic 650-kDa complex which binds 12(S)-hydroxy-5,8,10, 14-eicosatetraenoic acid (12(S)-HETE) with high affinity and specificity has been found in various cell lines but not until now in platelet cytosol. After incubation of human platelets with 12(S)-[3H]HETE, a labeled cytosolic 650-kDa complex was isolated. As previously shown for the binding complex in Lewis lung carcinoma (LLC) cells, ATP treatment transformed the platelet complex into a 50-kDa ligand-binding subunit. These results are of interest for two reasons: (a) 12(S)-HETE is a major arachidonic acid metabolite in platelets, and (b) platelets contain large amounts of the cell adhesion molecule GpIIb/IIIa, the activation of which is regulated by 12(S)-HETE. Hsp90 was found to be a component of the 12(S)-HETE binding complex in Lewis lung carcinoma cells, and the 50-kDa ligand-binding subunit itself bound 12(S)-HETE with high affinity. Competition experiments showed that 12(R)-HETE, 15 deoxy-Delta12, 14-prostaglandin J2, and 5(S)-HETE had lower affinity for the 50 kDa subunit than 12(S)-HETE. The 12(S)-HETE binding protein appears to be distinct from known members of the steroid hormone receptor superfamily of nuclear receptors. PMID- 10375397 TI - Characterization of point mutations in patients with pyruvate dehydrogenase deficiency: role of methionine-181, proline-188, and arginine-349 in the alpha subunit. AB - Human pyruvate dehydrogenase (E1), a heterotetramer (alpha2beta2), is the first component of the pyruvate dehydrogenase complex (PDC). E1 catalyzes the thiamin pyrophosphate (TPP)-dependent decarboxylation of pyruvate and the reductive acetylation of the dihydrolipoamide acetyltransferase component. Site-directed mutagenesis was employed to recreate three point mutations in the alpha subunit identified in E1-deficient patients, M181V, R349H, and P188L (P188A mutant E1 was used because of the very low level of expression of P188L), to investigate the functional roles of these three amino acid residues. P188A mutant E1 was much less thermostable than the wild-type E1. The kcats of M181V and P188A mutant E1s determined in the PDC reaction were 38 and 24% of that of the wild-type enzyme, respectively. The apparent Km for TPP for M181V increased significantly (approx 250-fold when determined in the PDC assay), while the apparent Km for pyruvate increased by only about 3-fold. In contrast, P188A had similar Kms for the coenzyme and the substrate as the wild-type. Km values for R349H were not determined due to the extremely low activity of this mutant (1.2% of the wild type E1-specific activity measured in the PDC assay). Wild-type E1 displayed a lag phase in the progress curve of the PDC reaction measured in the presence of low TPP concentrations (below 1 microM) only. All mutants had a lag phase that was not eliminated even at very high TPP concentrations, suggesting modifications in the conformation of the active site. Kinetic analysis indicated thiamin 2 thiothiazolone pyrophosphate (ThTTPP) to be an intermediate analog for wild-type human E1. M181V required a higher concentration of ThTTPP for inactivation than the wild-type and P188A E1s. The results of circular dichroism spectropolarimetry in the far UV region indicated that there were no major changes in the secondary structure of M181V, P188A, and R349H E1s. These mutant enzymes exhibited negative dichroic spectra at about 330 nm only in the presence of high TPP concentrations. This study suggests that arginine-349 is critical for E1's activity, methionine 181 is involved in the binding of TPP, and proline-188 is necessary for structural integrity of E1. PMID- 10375399 TI - Myeloperoxidase-catalyzed oxidation of tyrosine. AB - The oxidation of tyr by myleoperoxidase (MPO) is postulated to play a role in atherosclerotic plaque formation. MPO has been localized in plaques and a product of MPO-catalyzed oxidation of tyr, dityrosine, also found in plaques, is proposed to be a protein cross-linking agent. We have performed kinetic studies on the oxidation of tyr by MPO and investigated the role of substrate size on its oxidation. The kinetics of MPO-catalyzed oxidation of tyr where the tyr is free tyr, the dipeptides, tripeptides, and polypeptides were studied by stopped-flow methods. The rate of reaction with enzyme intermediates compound I and compound II are decreased with increasing substrate size. The amount of dityrosine formed was also decreased with increasing substrate size. The ability of sulfhydryl compounds to inhibit MPO-dependent dityrosine formation was investigated with reduced glutathione, cys, and met. Glutathione and cys both served as substrates for MPO compound I but not compound II, whereas met was not a substrate for either compound I or II. Met, an amino acid postulated to act as a "last chance" antioxidant for proteins, was not able to inhibit dityrosine formation from MPO catalyzed oxidation of tyr. Glutathione and cys caused partial inhibition; however, it is possible that this inhibition was due to their ability to react directly with MPO rather than trapping the tyr radicals. PMID- 10375398 TI - Phosphorylation and glycosylation of nucleoporins. AB - The nuclear pore complex mediates macromolecular transport between the nucleus and cytoplasm. Many nuclear pore components (nucleoporins) are modified by both phosphate and O-linked N-acetylglucosamine (O-GlcNAc). Among its many functions, protein phosphorylation plays essential roles in cell cycle progression. The role of O-GlcNAc addition is unknown. Here, levels of nucleoporin phosphorylation and glycosylation during cell cycle progression are examined. Whereas nuclear pore glycoproteins are phosphorylated in a cell-cycle-dependent manner, levels of O GlcNAc remain constant. The major nucleoporin p62 can be phosphorylated in vitro by protein kinase A and glycogen synthase kinase (GSK)-3alpha but not by cyclin B/cdc2 or GSK-3beta. The consensus sites of these kinases resemble sites which can be glycosylated by O-GlcNAc transferase. These data are consistent with a model that O-GlcNAc limits nucleoporin hyperphosphorylation during M-phase and hastens the resumption of regulated nuclear transport at the completion of cell division. PMID- 10375400 TI - Phosphatidylinositol 3-kinase regulates differentiation of H9c2 cardiomyoblasts mainly through the protein kinase B/Akt-independent pathway. AB - Phosphatidylinositol 3-kinase (PI3-kinase) is known to be a crucial regulator of muscle differentiation. However, its downstream pathway for this function is quite obscure. In this experiment we demonstrated the regulatory mechanism of the differentiation of H9c2 cardiomyoblasts, focusing on PI3-kinase, protein kinase B/Akt (PKB/Akt) and p42/44 mitogen-activated protein kinase (p42/44 MAPK). When H9c2 cells stably transfected with a constitutively active p110 (H9c2-p110*), a constitutively active PKB/Akt (H9c2-Akt), and an empty vector (H9c2-con) were induced to differentiate, H9c2-p110* cells differentiated fastest, followed by H9c2-Akt cells. H9c2-con cells differentiated at the slowest rate. Consistent with this result, LY294002 completely blocked differentiation of all these transfected cell lines, whereas PD098059 had no effect on their differentiation. When H9c2-p110* cells were transiently transfected with a dominant negative form of PKB/Akt, differentiation was not affected. Taken together, we concluded that PI3-kinase, but not p42/44 MAPK, regulates differentiation of H9c2 cardiomyoblasts mainly through the PKB/Akt-independent pathway. PMID- 10375402 TI - Phospholipase D activity of cytochrome P450 in human liver endoplasmic reticulum. AB - Phospholipase D (PLD) activity in mammalian liver endoplasmic reticulum (ER) has not been characterized. Purified human liver microsomal cytochromes P450 (P450) P450 1A2 and P450 2E1-were shown to have appreciable PLD activity, hydrolyzing phosphatidylcholine but not other phospholipids, generating PA and choline. The activity was confirmed using recombinant and mutated human P450s expressed in bacteria. In human liver microsomes, immunoinhibition of PLD activity was observed with anti-P450 1A2 > anti-P450 2C > anti-P450 2E1. Thus, P450 may act as a significant PLD in human liver ER and exert its biological effects by several mechanisms, including signaling functions and change of membrane properties. PMID- 10375401 TI - The extracellular loop between TM5 and TM6 of P-glycoprotein is required for reactivity with monoclonal antibody UIC2. AB - P-glycoprotein (P-gp), encoded by the MDR1 gene, is a plasma membrane transporter which confers resistance to many chemotherapeutic drugs. Monoclonal antibodies raised against P-gp have been used as tools to study P-gp topology and activity. Monoclonal antibody UIC2 recognizes a functional conformation of P-gp on the cell surface and blocks P-gp-mediated drug transport. Knowledge about the UIC2 epitope and the mechanism of its inhibitory effects may be helpful for understanding P-gp structure and developing P-gp inhibitors. In the present work, using several chimeras of MDR1 and MDR2, we found that the native sequence of the predicted extracellular loop between transmembrane domains (TM) 5 and 6 of P-gp is necessary, but not sufficient, for UIC2 reactivity. In addition, UIC2 reactivity is also affected by mutations in TM6, a region known to be involved in interactions of P-gp with substrates. These observations suggest that residues in the extracellular loop between TM5 and TM6 are directly involved in the display of the UIC2 epitope. Since TM6 has been shown to be actively involved in drug transport process, the proximity of this region to TM6 may help to explain why UIC2 binding is sensitive to the functional state of P-gp and why binding of UIC2 inhibits P-gp-mediated drug transport. PMID- 10375403 TI - GroEL-assisted and -unassisted refolding of mature and precursor adrenodoxin: the role of the precursor sequence. AB - We have performed refolding studies on a [2Fe-2S] protein, adrenodoxin (Adx), and its precursor form, preadrenodoxin. In vitro, mature Adx is expressed as a soluble active form in Escherichia coli, but precursor Adx is expressed in inclusion bodies. Both mature and precursor Adx refolded spontaneously from their denatured forms and the recovery levels of enzyme activities were 40 and 37% for mature and precursor Adx, respectively. Furthermore, the interaction between GroEL- and Gdn-HCl-denatured mature and precursor forms was investigated. In the case of mature Adx, the activity was increased in the presence of either GroEL, GroES, or bovine serum albumin and the refolding of mature Adx is a nonspecific process. However, the GroEL-mediated reaction is specific for precursor Adx under the experimental conditions used here. A higher electron transfer activity is obtained after ATP addition to the GroEL-containing refolding mixture, and GroEL precursor complexes were found by gel chromatography studies. Our observation suggests that the small single-domain protein Adx (mature form) folded independently of the chaperonin GroEL. The contribution of the chaperonin complexes to the folding is toward the aggregation-sensitive precursor Adx, which in vitro folded 1.3- to 1.4-fold slower than mature Adx. This demonstrates that the presequence is responsible for the formation of inclusion bodies and for the in vitro recognition motif for GroEL binding. PMID- 10375404 TI - Expression, purification, and in vitro characterization of the human outer mitochondrial membrane receptor human translocase of the outer mitochondrial membrane 20. AB - In order to investigate the biochemical properties of the mitochondrial outer membrane receptor, hTom20, involved in protein recognition, the cytosolic domain of this receptor was overexpressed and purified to homogeneity. A four-step purification including the purification of thrombin is described as well as an analysis of the function of the highly purified hTom20 protein. The receptor was concentrated and the subsequent aggregation behavior was investigated in order to understand the function of the single cysteine in the cytosolic domain as well as the function of the proposed "glutamine face" for the structure of the protein. It was found that specific dimerization of the cytosolic domain of hTom20 is necessary in order to prevent aggregation of the protein. In addition, the cysteine and the glutamine face are important for the stability of the protein. We propose that the function of the cysteine is to promote dimerization as found in the absence of dithiothreitol. PMID- 10375405 TI - A recombinant form of the catalytic subunit of phosphorylase kinase that is soluble, monomeric, and includes key C-terminal residues. AB - Residues 302-326 of the catalytic (gamma) subunit of phosphorylase kinase (PhK) may comprise an autoinhibitory, pseudosubstrate domain that binds calmodulin. To study this, the cDNA corresponding to rabbit muscle PhKgamma was expressed using Escherichia coli. This yielded two stable, high-activity PhKgamma forms (35 and 42 kDa by SDS-PAGE) that were smaller than an authentic sample of rabbit muscle PhKgamma (45 kDa by SDS-PAGE). Each recombinant form was purified to homogeneity. The N-terminal sequence of the larger, 42-kDa form (pk42) matched that of the rabbit muscle enzyme. This suggested that pk42 consisted of PhKgamma residues 1 362, including the putative calmodulin-binding, autoinhibitory domain. Kinetic parameters obtained for pk42 were like those previously reported for the intact gamma subunit. This implied that the lack of 25 PhKgamma C-terminal residues did not affect phosphorylase kinase activity, but greatly improved enzyme stability. An additional 60 residues were removed from the C-terminus of pk42 using the protease m-calpain. This increased the kinase activity 1.5-fold. Consistent with this, the activity of a mutant PhKgamma that consisted of residues 1-300, denoted gamma1-300, was like that of the m-calpain-treated enzyme. Therefore, although the effect was small, some influence by the C-terminus of pk42 was noted. Moreover, when pk42 was incubated with ATP alone, a C-terminal threonine residue became phosphorylated. Although the influence of this autophosphorylation cannot be inferred from this data, it was evidence that the C-terminus accessed the enzyme's active site. Taken together, these data imply that pk42 will be useful to study phosphorylase kinase structure/activity relationships. PMID- 10375406 TI - Degradation of chemicals by reactive radicals produced by cellobiose dehydrogenase from Phanerochaete chrysosporium. AB - Phanerochaete chrysosporium, grown on cellulose, produced a cellobiose-dependent dehydrogenase which reduced both ferric iron and molecular oxygen, resulting in the generation of the hydroxyl radical. The hydroxyl radical was detected in reaction mixtures with and without the addition of exogenous H2O2. The purified reductase and the fungus grown under nonligninolytic conditions that promote the production of the reductase were able to depolymerize an insoluble polyacrylate polymer. When oxalate, a secondary metabolite of P. chrysosporium, was used as the iron chelator, it was oxidized by the hydroxyl radical to form the carboxylate anion radical, a strong reductant. Under these reductive conditions, the enzyme was shown to catalyze the reduction of bromotrichloromethane to the trichloromethyl radical. We propose that these oxidative and reductive mechanisms may contribute to the degradation of a wide range of environmental pollutants by fungi which produce this enzyme. PMID- 10375407 TI - Extended heme promiscuity in the cyanobacterial cytochrome c oxidase: characterization of native complexes containing hemes A, O, and D, respectively. AB - The cyanobacteria Anacystis nidulans (Synechococcus sp. PCC6301), Synechocystis sp. PCC6803, Anabaena sp. PCC 7120, and Nostoc sp. PCC8009 were grown photoautotrophically under reduced oxygen tension in a medium with sulfate replaced by thiosulfate and nitrate replaced by ammonium as the S- and N-sources, respectively. In addition, Anabaena and Nostoc were grown under dinitrogen-fixing conditions in a medium free of combined nitrogen. Membranes were isolated from late-logarithmic cells (culture density corresponding to approximately 3 microliters packed cells per milliliter); cytoplasmic and thylakoid membranes were separated and purified according to established procedures. Acid-labile hemes were extracted from the membranes and subjected to reversed-phase high performance liquid chromatography. Separated hemes were analyzed spectroscopically and identified by comparison with authentic standards. In addition to hemes B, A, and O, the latter of which was induced under semianaerobic conditions only, substitution of thiosulfate and ammonium for the oxy-anions sulfate and nitrate led to the appearance of spectrally discernible heme D in the membranes and extracts therefrom. However, spectroscopic and kinetic investigation of the membrane-bound heme D rather disproved any reaction with oxygen or carbon monoxide. Kinetic measurements performed with the membrane bound respiratory oxidase gave evidence for only two kinetically competent terminal oxidases, a3 and o3, both apparently associated with a single type of apoprotein, viz. subunit I of the known cyanobacterial aa3-type cytochrome c oxidase. The heme D, on the other hand, seems to form a spectrally distinguished, yet kinetically ill-defined hemoprotein complex which does not qualify as a fully functional d-type terminal oxidase on our (wild-type) cyanobacteria even after growth under semianaerobic pseudo-reducing conditions. Also growth (of Anabaena and Nostoc) under dinitrogen-fixing conditions did not change this situation. Thus, we are left with (wild-type) cyanobacteria forming an unbranched respiratory chain with only a single type of terminal oxidase protein, viz. the known aa3-type cytochrome c oxidase. This oxidase, however, may incorporate different prosthetic (heme) groups in the sense of "heme promiscuity." Biosynthesis of the different heme groups thereby seems to respond to the ambient redox environment. In particular, however, conditions for expression of the two quinol oxidases potentially and additionally coded for by the genome of, e. g., Synechocystis sp. PCC6803 (see http://www.kazusa.or.jp/cyano), have not yet been found. PMID- 10375408 TI - Protein synthesis inhibitors and ethanol selectively enhance heterologous expression of P450s and related proteins in Escherichia coli. AB - The antibiotics chloramphenicol (Cm), tetracycline, and erythromycin, which inhibit bacterial protein synthesis and are known to induce the cold shock response, unexpectedly enhance the heterologous expression of P450s and related proteins in Escherichia coli. In contrast, antibiotics that mimic heat shock in E. coli such as puromycin, streptomycin, and kanamycin decrease the expression of the same proteins. A sublethal dose of Cm (1 microgram/ml) effectively enhances the expression of both membrane-bound proteins (microsomal and mitochondrial P450s) and a soluble mitochondrial protein (adrenodoxin) over the range of two- to eightfold. The expression level of N-terminal truncated P450c17 (1600 nmol/liter culture without Cm), for instance, reached 3500 nmol/liter culture by the addition of Cm, approximately 8.4% of the total cellular protein. Cm also enabled expression at useful levels of active P450s previously difficult to express in E. coli. In contrast, the expression of P450scc, a mitochondrial protein, is decreased by Cm but enhanced by ethanol, a powerful elicitor of heat shock response in E. coli. These results show that both the cold shock response induced by some antibiotics and the heat shock response induced by ethanol may lead to enhanced expression of certain heterologous proteins in E. coli. This study also indicates that protein synthesis inhibitors associated with the cold shock response may act as protein synthesis enhancers under certain conditions. PMID- 10375409 TI - The plasma membrane H+-ATPase from Tradescantia stem and leaf tissue is modulated in vitro by cGMP. PMID- 10375410 TI - Possible relationship between conditions associated with chronic hypoxia and brain mitochondrial DNA deletions; reduction of genomic 8-hydroxyguanine levels in human brain tissues containing elevated levels of the human mitochondrial DNA4977 deletion. PMID- 10375411 TI - Limited proteolysis of tyrosine hydroxylase identifies residues 33-50 as conformationally sensitive to phosphorylation state and dopamine binding. PMID- 10375412 TI - Molecular characterization of the enzyme catalyzing the aryl migration reaction of isoflavonoid biosynthesis in soybean. AB - The first specific reaction in the biosynthesis of isoflavonoid compounds in plants is the 2-hydroxylation, coupled to aryl migration, of a flavanone. Using a functional genomics approach, we have characterized a cDNA encoding a 2 hydroxyisoflavanone synthase from soybean (Glycine max). Microsomes isolated from insect cells expressing this cytochrome P450 from a baculovirus vector convert 4', 7-dihydroxyflavanone (liquiritigenin) to 4',7-dihydroxyisoflavone (daidzein), most likely via 2,4',7-trihydroxyisoflavanone which spontaneously dehydrates to daidzein. The enzyme also converts naringenin (4',5,7-trihydroxyflavanone) to genistein, but at a lower rate. 2-Hydroxyisoflavanone synthase transcripts are strongly induced in alfalfa cell suspensions in response to elicitation. PMID- 10375413 TI - Critical analysis of methods for assessment of predicted no-effect concentration. AB - Current risk assessment procedures for chemical substances are based on hazard quotients and require determination of a predicted no-effect concentration (PNEC). These concentrations are usually computed by applying an assessment factor to the lowest available toxicity value. However, other approaches have also been proposed based on statistical distribution of ecotoxicity data. This paper compares the various approaches in terms of precision and robustness. When only a few data are available, an assessment factor approach can be used. However, whenever possible (i.e. for a large set of chronic data), a statistical approach like that developed by T. Aldenberg and W. Slob (1991, Confidence Limits for Hazardous Concentrations Based on Logistically Distributed NOEC Toxicity Data, RIVM Report 71902002) with a 50% confidence level is the most precise and provides a stable value with increasing confidence when the number of tests increases. PMID- 10375414 TI - Transfers and transformations of zinc in flow-through wetland microcosms. AB - Two microcosm-scale wetlands (570-liter containers) were integratively designed and constructed to investigate transfers and transformations of zinc associated with an aqueous matrix, and to provide future design parameters for pilot-scale constructed wetlands. The fundamental design of these wetland microcosms was based on biogeochemical principles regulating fate and transformations of zinc (pH, redox, etc.). Each wetland consisted of a 45-cm hydrosoil depth inundated with 25 cm of water, and planted with Scirpus californicus. Zinc ( approximately 2 mg/liter) as ZnCl2 was amended to each wetland for 62 days. Individual wetland hydraulic retention times (HRT) were approximately 24 h. Total recoverable zinc was measured daily in microcosm inflow and outflows, and zinc concentrations in hydrosoil and S. californicus tissue were measured pre- and post-treatment. Ceriodaphnia dubia and Pimephales promelas7-day aqueous toxicity tests were performed on wetland inflows and outflows, and Hyalella azteca whole sediment toxicity tests (10-day) were performed pre- and post-treatment. Approximately 75% of total recoverable zinc was transferred from the water column. Toxicity decreased from inflow to outflow based on 7-day C. dubia tests, and survival of H. azteca in hydrosoil was >80%. Data illustrate the ability of integratively designed wetlands to transfer and sequester zinc from the water column while concomitantly decreasing associated toxicity. PMID- 10375415 TI - Expression of the vitellogenin gene in the liver of juvenile whitefish (Coregonus lavaretus L. s.l.) exposed to effluents from pulp and paper mills. AB - Juvenile whitefish (Coregonus lavaretus L. s.l.) were exposed by caging in the field to diluted effluents from three operating pulp, paper, and paperboard mills in Southern Lake Saimaa, Finland. The expression of the vitellogenin gene, used as a biomarker of estrogenic contamination of effluents, was measured using a Northern blotting method. Increased mRNA levels, the most specific and reliable evidence for estrogen receptor-mediated actions in vivo, were found in fish caged in the vicinity of one of three mills studied. This mill was found to discharge wood-derived compounds, such as sterols and resin acids, into Lake Saimaa in amounts considerably exceeding those from the other two mills. The increased vitellogenin gene expression suggests that the effluent is a source of estrogenic contaminants. PMID- 10375416 TI - Ecotoxicological characterization of energetic substances using a soil extraction procedure. AB - The acetonitrile-sonication extraction method (US EPA SW-846 Method 8330) and aquatic-based toxicity tests were used on laboratory and field samples, to characterize the ecotoxicity of soils contaminated with energetic substances. Spiked soil studies indicated that 2,4, 6-trinitrotoluene (TNT)-dependent soil toxicity could be measured in organic extracts and aqueous leachates using the 15 min Microtox (Vibrio fischeri, IC50=0.27 to 0.94 mg TNT/liter incubation medium) and 96-h Selenastrum capricornutum growth inhibition (IC50=0.62 to 1. 14 mg/liter) toxicity tests. Analyses of leachates of composite soil samples [containing TNT and some TNT metabolites, 1,3,5-trinitro-1,3, 5-triazacyclohexane (RDX), and 1,3,5,7-tetranitro-1,3,5, 7-tetrazacyclooctane (HMX)] from an explosives manufacturing facility, indicated toxicities similar to those found in the TNT-spiked soil studies and pure TNT in solution, and suggested that TNT was the major toxicant. Using TNT as a model toxicant in soils having different moisture contents (20% vs dry) and textures (sandy vs clayey-sandy) but similar organic matter content (3-4%), multi-factorial analyses of Microtox test data revealed that these soil factors significantly influenced the TNT extractability from soil and subsequent toxicity measurements. Taken together, data indicate that the modified Method 8330 may be used in conjunction with ecotoxicity tests to reflect the toxic potential of soils contaminated with energetic substances. PMID- 10375417 TI - Toxicity of organic and inorganic mercury to Saccharomyces cerevisiae. AB - In this study the effect if six different forms of mercury on the growth of the yeast Saccharomyces cerevisiae is presented. Five kinds of strains of S. cerevisiae were used. They were a wild type, a mercury-resistant type, and three mutants: mutation repair-deficient mutant, excision repair-deficient mutant, and recombination repair-deficient mutant. In terms of EC50 toward the wild-type strain, the toxicity order for the inorganic forms was Hg(NO3)2>HgSO4>HgCl2. Monovalent nitrate mercury Hg(NO3)2 was more toxic than bivalent Hg(NO3)2. The toxicity of organic mercury CH3HgCl on cell growth was two orders of magnitude higher than that of inorganic HgCl2. Between the two organic forms, CH3HgCl was more toxic than CH3HgOH. The survival rate in the presence of a certain concentration of CH3HgCl was about one-hundredth of the survival in presence of the same concentration of HgCl2. On the other hand, the concentration of CH3HgCl in the cell was about 170 times that of HgCl2. The addition of chelating agents, EDTA and methyl-penicillamine, to the medium did not reduce the toxicity of mercury. Among the three mutants tested, the one deficient in recombination repair systems was the most sensitive to mercury. PMID- 10375418 TI - Induction of metallothionein in dogwhelk Nucella lapillus during and after exposure to cadmium. AB - Induction of metallothionein (MT) was investigated in a common biomonitor, the dogwhelk Nucella lapillus (shell length: 27.7+/-1.4 mm; wet tissue weight: 667+/ 196 mg), during and after exposure to cadmium (Cd) under controlled laboratory conditions (10+/-1 degrees C and 34+/-1 per thousand salinity). The dogwhelks were exposed to 500 microg Cd l-1 (2.2% of 96 h LC50) for 60 days and then placed into clean seawater for 110 days. MT concentration in whole animal increased during the exposure period, peaked at Day 70, and then declined gradually. Half life of MT was ca. 40 days. MT concentration increased very significantly with increasing Cd concentration (r=0.74, n=24, P<0.001). Nevertheless, Cd concentration increased throughout the period of exposure and while in clean seawater, leveling off only after Day 120, indicating that Cd concentration could not be regulated by N. lapillus. Throughout the study, MT and Cd concentrations in gills, Leiblein gland, kidney, digestive gland, and gonad tissues increased gradually. Highest concentrations of MT and Cd were found in the Leiblein gland. Measurement of MT induction in the Leiblein gland of N. lapillus may therefore prove useful as a sublethal biological response to Cd contamination. PMID- 10375419 TI - The lethality of Salmosan (Azamethiphos) to American lobster (Homarus americanus) larvae, postlarvae, and adults. AB - The pesticide formulation Salmosan (47.5% w/w azamethiphos) is currently registered for use, in Canada, to treat salmonids for infestations of the copepod parasites, Lepeophtheirus salmonis and Caligus elongatus (sea lice). Determination was made of the acute lethality of this product to the three larval stages, the first postlarval stage, and the adult of the American lobster (Homarus americanus), a species of significant economic importance in Eastern Canada. The 48-h LC50 (as azamethiphos) is 3.57 microg/liter for Stage I, 1.03 microg/liter for Stage II, 2.29 microg/liter for Stage III, 2.12 microg/liter for Stage IV (the first postlarval stage), and 1.39 microg/liter for adults. These concentrations are not significantly different from each other, although the variability in response is greater in the larval stages than in the postlarvae or adults. These data when interpreted in conjunction with known physical oceanographic data and chemical dispersion studies indicate that single anti louse treatments are unlikely to result in mortality among lobsters in the vicinity of salmon farms. However, the sublethal effects of this product and the effects of repeated exposures have yet to be determined. PMID- 10375420 TI - Prediction of metal bioavailability in Dutch field soils for the oligochaete Enchytraeus crypticus. AB - Current risk assessment procedures ignore that variation in soil properties results in substantial differences for uptake and effects in organisms in different soils. In this contribution is presented the results of a study on the soil-related factors that modulate metal uptake and elimination by the oligochaete worm Enchytraeus crypticus. Uptake of Cd, Cu, Pb, and Zn was quantified in 20 Dutch field soils as a function of time. Uptake rate constants and equilibrium concentrations were estimated using compartment modeling. Internal metal concentrations varied less than the corresponding external levels. Zn and especially Cu provided the most extreme examples of this general behavior, which suggests regulation by the organism. Body residues by Cd increased linearly over time in 11 of the 20 soils studied, whereas in the remaining 9 soils equilibration of internal Cd levels was observed. CaCl2 extraction could be used to discriminate the 9 soils in which there is Pb accumulation from the 11 soils in which bioavailable Pb levels were too low to allow for uptake. Multivariate expressions that describe uptake rate constants and bioaccumulation factors as a function of soil characteristics were derived. pH and cation exchange capacity were the most important parameters. The formulae were very similar to those describing partitioning of metals over the solid and liquid phase of the soils, which suggests pore water-mediated uptake. A semi-mechanistic approach yielded further evidence of pore water-related uptake, modulated by competition between H+ and metal ions at the active sites of the membranes. PMID- 10375421 TI - Early expression of ubiquitin in myofibers of rats in organophosphate intoxication. AB - The degenerative process of the myofibers of the diaphragm of rats intoxicated with the organophosphate isofenphos, a compound that inhibits esterases, was studied at different intervals of intoxication. Early disorganization of the intermyofibrillar network and of the myofilaments, as well as dilatation of organelles, were observed by use of transmission electron microscopy. These changes precede macrophage invasion of the muscle fibers. Early expression of ubiquitin was observed in segments of muscle fibers by immunohistochemistry. Bands of polyubiquitin complexes in muscle homogenates were observed by immunoblotting. These bands disappeared in later stages of intoxication. A 42.5 kDa band corresponds to actin, as observed by immunoblotting using antisarcometric actin. This indicates relatively large amounts of polyubiquitin complex associated with sarcomeric actin in muscle fibers in early stages of intoxication. Based on these results it seems that actin is an important target in organophosphate-induced myofiber degradation and that the degradation of this protein-by the polyubiquitin pathway-may play an important role in the early disorganization of the sarcomere, as observed by electron microscopy. A possible role of the ubiquitin proteolytic pathway is that of trying to eliminate proteins modified in the early phases of muscle fiber degeneration, which is a necessary step for regeneration of the posterior segmental muscle. PMID- 10375422 TI - Effects of polychlorinated biphenyls on thyroid hormones and liver type I monodeiodinase in the chick embryo. AB - Polychlorinated biphenyls (PCBs) are widespread environmental contaminants which can biomagnify to higher tropic level organisms including birds. Circulating thyroid hormones (TH) and growth are decreased by PCB exposure. The first set of studies investigated the effects of PCBs on an enzyme responsible for TH homeostasis, hepatic type I monodeiodinase (MDI) in chicken embryos. Fertile chicken eggs were injected with Aroclor 1242, Aroclor 1254, 2,2',6, 6' tetrachlorobiphenyl (TCB), 3,3',4,4'-TCB, or 3,3',5,5'-TCB on Day 0 and studies were terminated on Incubation Day 21. Hepatic MDI activity was reduced in embryos treated with the Aroclor mixtures. No effects on MDI activities were observed after PCB isomer treatment. Liver weights from embryos treated with Aroclor 1242 were decreased. In the second study, chick embryos were exposed to these same PCBs in order to evaluate their effect on circulating THs and growth. Treatment with PCBs had no effect on body weight. Femur length were decreased with Arcolor 1242 treatment. A decrease in plasma concentration of thyroxine was observed after treatment with Aroclor 1242 and Aroclor 1254. Based on these findings, it is evident that PCBs alter the thyroid axis. Bird circulating TH levels, which are generally reported, may not be a good biomarker for low-dose exposure to PCBs. However, the reduction in MDI activity was more sensitive to PCB mixture exposure and may be a useful biomarker. PMID- 10375423 TI - Impacts of structural photomodification on the toxicity of environmental contaminants: anthracene photooxidation products. AB - The toxicity of polycyclic aromatic hydrocarbons (PHAs) is known to be enhanced by light via photosensitization reactions (production of active oxygen) and photomodification of the chemicals (e.g., oxidation) to more toxic compounds. Anthracene (ANT) toxicity in particular has been found to increase dramatically following photomodification. The objective of this study was to identify the photooxidation products of ANT and assess the toxicity of selected photoproducts. High performance liquid chromatography (HPLC) analysis of anthracene photooxidation revealed a complex array of oxidation products; prevalent among these were anthraquinone (ATQ) and hydroxy-anthraquinones (hATQs). Eleven of these compounds were tested for toxicity using growth inhibition of the duckweed Lemna gibba L. G-3. All but one of the compounds tested were found to be toxic, and when UV radiation was present in the light source toxicity was generally enhanced. The chemicals were also irradiated under SSR prior to toxicity testing. In about half the cases, the ATQ compounds were rapidly photooxidized and the resultant photoproducts were more toxic than the parent compounds. Interestingly, 2-hydroxyanthraquinone, which was not subject to photooxidation, was the most toxic of the compounds tested. As a light stable compound it presents the risk of a persistent environmental hazard. PMID- 10375424 TI - Effects of trifluralin on carp: biochemical and histological evaluation. AB - Acute and subacute toxicity of the herbicide trifluralin on fish was investigated in laboratory toxicity tests with carp. Median lethal concentrations were determined in acute tests. The 96-h LC50 value was 0.045 (0.036-0.051)mg/L. Fish were exposed to subacute concentrations of the herbicide (0.005, 0.01, and 0.02 mg/L trifluralin) in the 14-day toxicity tests and the effects on the relative growth rate, some biochemical parameters [alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanin aminotransferase (ALT) activities in serum, gills, liver, and kidney], gills, liver, and kidney structure were studied. A decrease in relative growth rate was found. An increase of functional enzyme activities in blood serum and the organs examined, particularly in the highest concentration of trifluralin indicated changes in the vital organs, was confirmed by histological analysis. The most severe changes (although mostly reversible) were found in the gills and kidney of the fish examined. PMID- 10375426 TI - Environmental research, section A PMID- 10375425 TI - Validation of genetically engineered bioluminescent surfactant resistant bacteria as toxicity assessment tools. AB - Bacteria are useful organisms for measuring acute and chronic toxicity. The most popular toxicity tests utilize the inhibition of bioluminescence as an indication of toxicity. An extensive toxicity database on pure chemical compounds has been created using the bioluminescent microorganism, Vibrio fischeri. However, the use of the Microtox assay in applications for environmental samples is not always successful, due to the test organism. Because the genes for bioluminescence have been cloned from V. fischeri, environmentally relevant test strains can be readily constructed. In this study, surfactant-resistant bioluminescent bacterial strains were constructed by transferring a broad host range plasmid containing the bioluminescent genes under the regulation of a constitutive promoter into strains from several bacterial genera. Two test strains, Stenotrophomonas 3664 and Alcaligenes eutrophus 2050, were approximately 400 times more resistant to the nonionic surfactant polyoxyethylene 10 lauryl ether than V. fischeri and are useful for toxicity reduction evaluations of remediation processes which use surfactants for solubilization of hydrophobic pollutants. The use of these strains as alternative test organisms in the Microtox assay was evaluated using nonpolar narcosis as the baseline toxicity mechanisms. The two test strains and V. fischeri indicated linear fits of EC50 values with the octanol/water partition (Kow) for five nonpolar narcotic compounds in acute assays (r2>0.9) with a slope of approximately 1. For all three strains, the y-intercept values were approximately the same, indicating that sensitivity did not vary. These results indicate that the nonpolar narcosis baseline toxicity mechanism may be useful as a general tool to validate the functioning of genetically engineered bioluminescent microorganisms. PMID- 10375427 TI - Combined effects of H-7 and cytochalasin B on outflow facility in monkeys. AB - The serine-threonine protein kinase inhibitor H-7 and the fungal metabolite cytochalasin B (CB) disrupt the actin microfilament network by different mechanisms, and increase outflow facility similarly in live monkeys. Their combined effect has therefore determined on total outflow facility in cynomolgus monkeys by 2-level constant pressure perfusion. (1) After unilateral anterior chamber (AC) bolus injection of H-7 [10 micrometers, 100 micrometers (subthreshold for increasing facility when given alone) or 500 micrometers (just threshold)] followed by bilateral AC bolus injection of CB [2 micrograms (strong but submaximal for increasing facility when given alone)], no significant difference between eyes was observed. (2) After bilateral AC exchange with a subthreshold dose of H-7 (10 micrometers) followed by unilateral AC bolus injection of a subthreshold dose of CB (0.02, 0.05, 0.1 or 0.5 microgram), 10 micrometers H-7 plus 0.1 or 0.5 microgram CB increased facility by approximately 40 or 80% compared to 10 micrometers H-7 alone. (3) After bilateral AC exchange with a maximal dose of H-7 (300 micrometers), followed by unilateral AC bolus injection of a subthreshold dose of CB (0.1 or 0.5 microgram), 300 micrometers H 7 plus 0.5 microgram CB increased outflow facility by 47% compared to 300 micrometers H-7 alone. (4) After unilateral AC exchange with a maximal dose of H 7 (300 micrometers) followed by bilateral AC bolus injection of a near-maximal dose of CB (2 micrograms), 300 micrometers H-7 plus 2 micrograms CB increased the facility by 67% compared to 2 micrograms CB alone. The significant effect of combined subthreshold doses of H-7 and CB on outflow facility, the potentiation of the facility-increasing effect of a maximal H-7 dose by both subthreshold and near-maximal CB doses, and the known cytoskeletal effects of both compounds, may suggest that both increase facility by disrupting actin filaments in the trabecular meshwork. PMID- 10375428 TI - Retinochoroiditis is induced by oral administration of Toxoplasma gondii cysts in the hamster model. AB - Administration of Toxoplasma cysts by intraperitoneal innoculation in the Syrian Golden Hamster provides a reproducible animal model of acquired Toxoplasmic retinochoroiditis and cysts are observed in the brain. However, toxoplasmosis is frequently acquired by oral ingestion of contaminated foodstuffs and it is recognised that the route by which disease is acquired may influence its pathogenesis and clinical expression. This study aimed to determine whether retinochorioiditis and cysts in the brain develop after oral ingestion in the Syrian Golden Hamster model as this is the route of induction akin to that in man and may therefore be more relevant in the study of disease pathogenesis. All animals developed disease by 4 weeks. Ocular and cerebral inflammation was confirmed by histology at 16 weeks and this was milder than in the original model. PMID- 10375429 TI - The influence of eye solutions on blinking and ocular comfort at rest and during work at video display terminals. AB - The aim of this work was to study blink frequency changes and levels of ocular discomfort during work at a video display terminal, and the effects on these parameters of augmented or reduced humidification of the ocular surface. Blink rate was measured from recordings of the electrical signal evoked by the contraction of the orbicularis oculi muscle. Blink rate and interblink intervals were analyzed at rest and during performance of a task with a computer (playing a card game) for 10 or 30 min in steady environmental conditions and during application of a continuous stream of air to the face. In two separate sessions, the effect of pretreatment with humidifying ocular solutions of different elastoviscosity (balanced salt solution or elastoviscous 0.1% Hylan A solution) was assayed. At the end of each experimental period, the subjects marked the level of ocular discomfort experienced on a 0-10 cm visual analogue scale. The blink frequency at rest (12.4+/-1.2 blinks min-1) was reduced significantly (to 10.3+/-1.1 blinks min-1) by pretreatment with elastoviscous eyedrops both with and without air applied to the face. This effect was not obtained with balanced salt solution. During performance of the visual task for 10 or 30 min, basal blink rate decreased significantly, to about 40% of the control value. Neither application of an air jet on the face nor application of eye solutions of different viscosity modified this reduced blink rate.A low degree of ocular discomfort developed after performance of the visual task that was enhanced by air application to the face. This discomfort was reduced by pretreatment with ocular solutions, the elastoviscous eye solution being more efficient than the balanced salt solution. Interblink interval duration was also more regular after treatment with the elastoviscous solution. These data suggest that blink rate at rest is maintained in part by activation of sensory receptors of the cornea and conjunctiva, which are stimulated by desiccation of the ocular surface. Reduction of eye blink frequency elicited by the performance of a visual task with a computer appears to depend on central neural mechanisms that are quite independent of peripheral sensory inputs. The reduction of blink frequency consecutive to computer use was associated with a sensation of discomfort that was attenuated more effectively by elastoviscous eyedrops than by regular balanced salt solution. PMID- 10375430 TI - Evaluation of the human choroidal melanoma rabbit model for studying microcirculation patterns with confocal ICG and histology. AB - The aim of this study was to develop consistently focal elevated choroidal masses of human choroidal melanoma in immunosuppressed rabbits and to correlate the visualization of prognostically significant microcirculation patterns from confocal indocyanine green angiography with histologic microcirculation patterns. A human choroidal melanoma cell line (OCM1) was implanted in the choroid of 40 rabbit eyes using three different techniques: transscleral choroidal injection of a cell suspension, injection of a cell suspension in a surgically induced cyclodialysis cleft, and implantation of solid tumor fragments in a surgically induced cyclodialysis cleft. The rabbits were immunosuppressed with daily injections of Cyclosporin A to prevent host versus graft reaction. The eyes were studied weekly with indirect ophthalmoscopy and fundus photography to monitor tumor growth and indocyanine green angiography using a confocal scanning laser ophthalmoscope to identify microcirculation patterns in vivo and correlate these findings with the histologic demonstration of tumor microcirculation patterns. A tumor mass was identified by indirect ophthalmoscopy in 16 of the 40 implanted rabbit eyes (40%). Each of these tumors was confirmed histologically to represent a focal elevated choroidal mass. All 16 elevated choroidal masses grow in eyes in which solid tumor fragments were implanted. In total, a melanoma was identified histologically in 28 of the implanted 40 eyes (70%). In addition to the 16 eyes where the melanoma appeared as a focal elevated choroidal mass, 4 eyes contained a focal elevated mass in the sclera and 8 eyes contained a flat choroidal tumor. Histologically, microcirculation patterns were identified only in the 16 eyes with focal elevated choroidal masses. Confocal indocyanine green angiography imaged microcirculation patterns in 13 of these 16 eyes (81%). The surgical implantation of small solid fragments of human choroidal melanoma in immunosuppressed rabbit eyes provides the best method to consistently obtain focal elevated choroidal masses. These focal elevated choroidal masses resemble booth the localization and the growth pattern of choroidal melanomas in humans. In addition, they also contain microcirculation patterns similar to those seen in humans that are detectable with confocal indocyanine green angiography. The use of indocyanine green angiography with this animal model may be especially useful in designing and evaluating anti-microcirculation treatments directed at uveal melanoma. PMID- 10375431 TI - Epitope discovery using bacteriophage display: the minimum epitope of an anti IRBP antibody. AB - The purpose of this study was to determine, using random peptide library (RPL) technologies, the minimal epitope requirements of the mouse monoclonal anti interphotoreceptor-retinoid-binding protein antibody, H3B5. This previously characterized antibody is used as an example to examine whether RPL's offer a relatively easy and rapid route to obtaining detailed epitope mapping data.A pentadecamer random peptide library (RPL) displayed on the major coat protein (gene 8) of filamentous bacteriophage (F88-4-15) was used as a target for selection by the anti-IRBP monoclonal antibody, H3B5. Three rounds of library selection were performed, and 90 of the resultant RPL clones were examined for affinity to H3B5 by enzyme-linked immunosorbent assay (ELISA). DNA sequencing of ELISA positive clones provided sequence of the region encoding the random peptide. After three rounds of selection of the RPL, 76.7% of clones examined interacted with H3B5, 17.7% did not show significant binding and 6.6% bound to control antibody also. The essential elements of the peptide epitope were determined by sequence comparison of 24 clones to be the four amino-acid sequence (Aspartic or glutamic acid)-Proline-Arginine-(Leucine, Isoleucine or Valine). This motif [(D/E) PR (L/I/V)] is in agreement, but at greater resolution, than previous synthetic peptide studies where the motif AASEDPRL was identified. Other motifs were found which bound to H3B5 but did not share primary structure similarities (peptidomimetics). Selection from a RPL has rapidly defined the minimal requirements for the H3B5 epitope in fine detail. Such a process offers great potential for investigating antibody-antigen interactions and core sequences of an epitope, and enables the identification of motifs in other proteins which may be recognized by the antibody, providing information on possible cross-reactivity. PMID- 10375432 TI - Vasoactive intestinal polypeptide (VIP) innervation of the human eyelid glands. AB - This study was conducted to obtain morphological proof of innervating nerve fibres in the glands of the human eyelid (accessory lacrimal glands of Wolfring, meibomian glands, goblet cells, glands of Zeis, glands of Moll, sweat glands, glands of lanugo hair follicles) and identification of the secretomotorically active neuropeptide vasoactive intestinal polypeptide (VIP) as a common transmitter. Epoxy-embedded ultrathin sections of tissue samples from human eyelids were studied using electron microscopy. Paraffin sections fixed in Bouin Hollande solution were immunostained with rabbit antiserum against VIP. With the electron microscope we were able to identify nerves in the glandular stroma of all the glands examined with the exception of goblet cells. Intraepithelial single axons were only seen in the parenchyma of Wolfring glands. The morphological findings corresponded with the immunological finding of VIP positive, nerve-like structures in the same locations, with the exception of lanugo hair follicle glands, and goblet cells. Our findings indicate that the glands of the eyelids and main lacrimal gland represent a functional unit with VIP as a possible common stimulating factor. PMID- 10375433 TI - Antioxidant activity of retinol, glutathione, and taurine in bovine photoreceptor cell membranes. AB - The antioxidant activities of compounds endogenous to mammalian rod outer segments (ROS) were investigated in vitro by measuring the oxidative loss of polyunsaturated fatty acids (PUFA's) from the membranes of intact ROS and from liposomes made from ROS phospholipids (PL) to which lipid soluble compounds had been added. The membranes were exposed to the water-soluble oxidant 2, 2' azobis(2-amidinopropane) dihydrochloride (AAPH). Retinol protected PUFA's in ROS liposome PL's, whereas retinaldehyde promoted lipid peroxidation. When isolated ROS were stimulated to produce endogenous retinol, PUFA loss was inhibited by up to 17%. These findings suggest an antioxidant function for the enzymatic reduction of retinaldehyde to retinol during the visual cycle. Water-soluble antioxidants, taurine and reduced glutathione (GSH), were investigated individually and in combination with retinol in ROS PL liposomes. GSH protected PUFA's in ROS PL liposomes. Taurine alone showed little antioxidant activity, but in combination with retinol it protected lipids twice as much as retinol alone. These results support previous findings that taurine protects ROS lipids during exposure to cyclic light. PMID- 10375434 TI - Regulation of calcium/calmodulin-dependent protein kinase II in the adult rat retina is mediated by ionotropic glutamate receptors. AB - This study is concerned with the transmitter-mediated regulation of the alpha(50 kDa) and beta(60 kDa) subunits of calcium calmodulin dependent protein kinase II (CamKII) in the adult rat retina. The level of antibody binding to the CamKII and the activity of CamKII were found to be increased after intravitreal injection of glutamate. Changes in the levels of the antibody-binding to the subunits of CamKII were observed in different subcellular fractions of the retina with a maximum response observed in crude synaptic membrane fractions. The glutamate mediated increases in CamKII were specific and blocked by 3,5-Dimethyl-1 adamantanamine; 3,5-Dimethylamantadine (Memantine), (+/-) 2-Amino-5 Phosphopentonic (AP-5) and 6-Cyano-7-Nitroquinoxaline-2,3-Dione (CNQX) but not with dl -2-Amino-3-Phosphono-Propionic (AP-3). The results indicate that the retinal neurotransmitter, glutamate, can regulate retinal CamKII activity through ionotropic but not metabotropic glutamate receptors. NMDA-receptors were found to be necessary but insufficient to stimulate CamKII. A model in which cooperative interaction between NMDA and non-NMDA glutamate receptors/ion channels is presented to explain the glutamate stimulated increases in CamKII activity in the retina. PMID- 10375435 TI - Does glutathione-S-transferase dethiolate lens protein-thiol mixed disulfides?-A comparative study with thioltransferase. AB - Protein S-thiolation is a process in which under oxidative stress, vulnerable sulfhydryl groups of proteins are conjugated to non-protein thiols such as glutathione (GSH) or cysteine resulting in the formation of protein-thiol mixed disulfides, protein-S-S-glutathione (PSSG) and protein-S-S-cysteine (PSSC). This process spontaneously disrupts the redox homeostasis of the cells, which in turn leads to functional disturbances in the respective tissue. In the ocular lens, such modification of proteins may trigger a cascade of events starting with the alteration of protein conformation, protein/enzyme deactivation, protein-S-S protein aggregation and eventually lens opacification or cataract. Generally, the first line of defense system in the cells protects the lens proteins against such damage. Recent studies in our laboratory have shown that in addition to this defense system, lens cells also possess a well developed system to repair the oxidative damage to the lens proteins. We have identified this repair system as thioltransferase (TTase) and have proved that TTase by its dethiolase activity reverses the protein S-thiolation process which returns the oxidatively damaged lens proteins/enzymes to their original reduced state and restores their physiological functions. We investigated if this repair mechanism was mediated by enzymes other than TTase. We studied glutathione S-transferase (GST) and report here for the first time the cloning, high level expression, and purification of human lens mu and pi isoforms of GST. A comparative study of recombinant human lens TTase and GST (mu and pi) on their dethiolating abilities using lens crystallin-thiol mixed disulfides showed that the lens TTase is 60-70% more efficient in the dethiolation/repair process than GST. When TTase and GST were tested in conjunction for the dethiolation of thiol mixed disulfides, there was no significant enhancement of dethiolase activity. These findings suggest that TTase by itself is an efficient enzyme in the dethiolation/repair process and hence can be considered a crucial system to counteract oxidative stress in the lens. PMID- 10375436 TI - Identification and immunolocalization of actin cytoskeletal components in light- and dark-adapted octopus retinas. AB - Photoreceptors in the octopus retina are of the rhabdomeric type, with rhabdomeres arising from the plasma membrane on opposite sides of the cylindrical outer segment. Each rhabdomere microvillus has an actin filament core, but other actin-binding proteins have not been identified. We used immunoblotting techniques to identify actin-binding proteins in octopus retinal extracts and immunofluorescence microscopy to localize the same proteins in fixed tissue. Antibodies directed against alpha-actinin and vinculin recognized single protein bands on immunoblots of octopus retinal extract with molecular weights comparable to the same proteins in other tissues. Anti-filamin identified two closely spaced bands similar in molecular weight to filamin in other species. Antibodies to the larger of the Drosophila ninaC gene products, p174, identified two bands lower in molecular weight than p174. Anti-villin localized a band that was significantly less in molecular weight than villin found in other cells. Epifluorescence and confocal microscopy were used to map the location of the same actin-binding proteins in dark- and light-adapted octopus photoreceptors and other retinal cells. Antibodies to most of the actin-binding proteins showed heavy staining of the photoreceptor proximal/supportive cell region accompanied by rhabdom membrane and rhabdom tip staining, although subtle differences were detected with individual antibodies. In dark-adapted retinas anti-alpha-actinin stained the photoreceptor proximal/supportive cell region where an extensive junctional complex joins these two cell types, but in the light, immunoreactivity extended above the junctional complex into the rhabdom bases. Most antibodies densely stained the rhabdom tips but anti-villin exhibited a striated pattern of localization at the tips. We believe that the actin-binding proteins identified in the octopus retina may play a significant role in the formation of new rhabdomere microvilli in the dark. We speculate that these proteins and actin remain associated with an avillar membrane that connects opposing sets of rhabdomeres in light-adapted retinas. Association of these cytoskeletal proteins with the avillar membrane would constitute a pool of proteins that could be recruited for rapid microvillus formation from the previously avillar region. PMID- 10375437 TI - Histamine effects on conjunctival fibroblasts from patients with vernal conjunctivitis. AB - Histamine, an important mast cell mediator in allergic disorders, may affect extracellular matrix production and cell growth in vernal keratoconjunctivitis (VKC). In the present study, the histamine reactivity of conjunctival fibroblasts derived from VKC patients was investigated in vitro. Conjunctival fibroblast cultures were derived from biopses of 8 tarsal VKC patients and 5 normal subjects. These cells were maintained in vitro and stimulated with different concentrations of histamine with and without H1 (clorpheniramine) and H2 (cimetidine) receptor antagonists. Comparisons were made to fibroblasts grown in the same media without histamine and to fibroblasts stimulated with just antihistamine. The effects of histamine were evaluated by: (1) the MTT test to assess cell proliferation; (2) an in vitro wound model for cell migration and (3) the measurement of procollagen I (PIP) and procollagen III (PIIIP) in supernatants for collagen production. Results showed: (1) While VKC-derived fibroblasts proliferated at a faster rate than normal cells in unstimulated media, after histamine stimulation, VKC and normal cells grew at a similar rate. Both H1 and H2 antagonists significantly inhibited (P<0.05) histamine-induced cell proliferation. (2) Histamine enhanced cell migration after wounding; this effect was inhibited only by H2 antagonism. (3) When stimulated with histamine, VKC fibroblasts produced significantly more PIP than those in control media. Furthermore, VKC-derived fibroblasts were more sensitive to histamine challenge, producing significantly more PIP than normal fibroblasts. H1 and H2 antagonists did not modify histamine-stimulated PIP production. The enhanced proliferative and productive capacity of VKC fibroblasts may be the result of a selective overgrowth of one or more fibroblast subpopulations in a chronically inflamed tissue. Histamine increased proliferation, migration and collagen production in both normal and VKC fibroblasts. Since H2 antagonism modulated both cell growth and migration, but not histamine-induced collagen production, the latter may be mediated by a different receptor. These results showed that histamine is at least partially responsible for fibroblast stimulation. PMID- 10375438 TI - Preventive effect of topical vitamin E-containing liposome instillation on the progression of galactose cataract. Comparison between 5-week- and 12-week-old rats fed a 25% galactose diet. AB - The preventive effect of topical vitamin E-containing liposome instillation on the progression of galactose cataract was compared between 5-week- and 12-week old Wistar rats fed a 25% galactose diet. Vitamin E-containing liposomes [LP(+VE)] and vitamin E-free liposomes [LP(-VE)] were prepared with dipalmitoylphosphatidylcholine and dioleoylphosphatidylcholine (7:3 w/w). Twice daily instillation of either LP(-VE) or LP(-VE) into both eyes of 5-week-old rats fed the galactose diet for 18 days (5WGR) and 12-week-old rats fed the galactose diet for 7 weeks (12WGR) at which time some vacuoles appeared in the lens cortical equator, was conducted for a period of 4 and 9 weeks, respectively. The severity of cataracts at the end of instillation was similar in 5WGR and 12WGR. Instillation of LP(+VE), but not LP(-VE), retarded cataract progression in 5WGR and 12WGR. In 12WGR, LP(-VE) instillation caused a transient retardation of the progression. In lenses of 5WGR and 12WGR, decreases in vitamin E and reduced glutathione contents and increases in lipid peroxide, galactitol, and water contents occurred at the onset of instillation. For 5WGR, a decrease in lens reduced glutathione content and increases in lens vitamin E, lipid peroxide, galactitol, and water contents occurred at the end of instillation. For 12WGR, decreases in lens reduced glutathione and vitamin E contents and increases in lens lipid peroxide, galactitol, and water contents occurred at the end of instillation. In sera of 5WGR and 12WGR, vitamin E concentration decreased at the onset of instillation increased at the end in 5WGR and was unchanged in 12WGR. In 5WGR, instillation of LP(+VE), but not LP(-VE), for 4 weeks prevented these changes except the changes in lens galactitol and water contents and serum vitamin E concentration. In 12WGR, instillation of LP(+VE), but not LP(-VE), for 9 weeks prevented these changes except the changes in lens galactitol and water contents and serum vitamin E concentration. These results indicate that topically instilled LP(+VE) can retard cataract progression in 5WGR and 12WGR, mainly by the antioxidative action of vitamin E contained in the instilled liposomes. PMID- 10375439 TI - Influence of UVA light stress on photoreceptor cell metabolism: decreased rates of rhodopsin regeneration and opsin synthesis. AB - There is considerable evidence indicating that rhodopsin is the chromophore mediating light damage to the rat retina caused by exposure to mid-visible wavelengths. Retinal damage is, however, more effectively produced by short wavelength light, and little is known about the initiating events for this damage class. The present study sought to determine the involvement of rhodopsin bleaching in short-wavelength damage by examining rhodopsin levels and opsin synthesis at early time points following acute ultraviolet-A (UVA) exposures of the pigmented rat eye. A gradual decline in rhodopsin to 8% of the level in non exposed control eyes occurred over a 1 hr exposure to 1500 microW cm-2of UVA light. When animals were placed in darkness following this exposure, rhodopsin had recovered to only 27% of control levels by 2 hr post-exposure indicating a very slow rate of regeneration. For later time points, animals were returned to dim cyclic light and by 2 days following exposure, rhodopsin levels had risen to 57% of control. In contrast, opsin levels at this same time point were unaffected by UVA exposure. Other observations indicating the UVA exposure affected photoreceptor cell metabolism included a 27% decrease in the rate of opsin synthesis between 1 and 2 days following exposure, and a 69% reduction in the rate of rod outer segment disk renewal during the initial 3 days following exposure. These data show that UVA light stress in the retina causes a gradual bleaching of rhodopsin followed by a slow rate of recovery and altered photoreceptor cell metabolism. These results are consistent with the concept that rhodopsin mediates UVA-induced retinal damage and the possible mechanisms by which this might occur are discussed in relation to alternative hypotheses currently in the literature. PMID- 10375440 TI - Hydrogen peroxide causes significant mitochondrial DNA damage in human RPE cells. AB - Retinal pigment epithelial cell dysfunction mediated by reactive oxygen intermediates has been suggested as a possible cause of age-related macular degeneration. To test the hypothesis that retinal pigment cells are susceptible to genetic damage mediated by reactive oxygen intermediates, retinal pigment epithelial cells were treated with 50 micrometers-200 micrometers of hydrogen peroxide in vitro. Damage to mitochondrial DNA and three nuclear loci were assessed using quantitative polymerase chain reaction. Hydrogen peroxide treatment of retinal pigment epithelial cells resulted in significantly increased mitochondrial DNA damage. Significant mitochondrial DNA damage occurred rapidly and was not completely repaired within 3 hr post-treatment. By contrast, no DNA damage was observed in three different nuclear loci (beta-globin gene cluster, hprt, and beta- polymerase genes). Hydrogen peroxide treatment of retinal pigment epithelial cells also resulted in decreased mitochondrial redox function compared to controls, consistent with increased mitochondrial DNA damage. Consequently, retinal pigment epithelial cell mitochondrial DNA appears susceptible to hydrogen peroxide mediated damage in vitro, and thus, may serve as a catalyst in the initial events leading to retinal pigment epithelial cell dysfunction in vivo. PMID- 10375441 TI - Structure and stability of the dityrosine-linked dimer of gammaB-crystallin. AB - Oxidative damage to proteins leads to a variety of modifications such as racemization, carbonyl compound formation, new fluorophores, aggregation, crosslinking and insolubility, several of which are markers of pathogenesis. A particular modification that has been associated with abnormal and pathological situations is the dityrosine crosslink in proteins, thought to be responsible for the reduced solubility and elasticity of proteins, and plaque formation. Dityrosine crosslinking has been suspected to occur in the crystallins of the eye lens during cataract. We focus attention here on the generation, structure and conformational stability of such a dityrosine-linked protein of the eye lens. We find this crosslink to be readily generated photodynamically in the presence of sensitizers. Among the crystallins, crosslinking occurs most readily in the gamma crystallins under these conditions. We have isolated, purified and studied the properties of the dityrosine-linked dimer of the eye lens protein gammaB crystallin. While the dityrosine crosslink does not alter the secondary structure of the protein, it changes the tertiary structure in a subtle manner. This alteration destabilizes the dimer, which denatures more readily than the parent monomer, and also makes it precipitate more readily, a point of relevance to cataractogenesis of the eye lens. Comparison of these results with those reported on other dityrosine-dimerized proteins suggests that while the conformation of these proteins might not be altered in a major manner upon dityrosine linkage, the dimer is structurally less stable and displays reduced solubility, both of which are of pathological importance. PMID- 10375442 TI - The optical properties of the anterior segment of the eye: implications for cortical cataract. AB - Epidemiological studies have correlated cortical cataract with exposure to light and have suggested that this is due primarily to relatively short wavelengths of ultraviolet radiation (UV-B). In addition, some cellular and animal models also implicate UV-B. In order to evaluate the likely role of different wavelengths of light in the etiology of cortical cataracts, the optical characteristics of several animal models were ascertained and compared to the primate. This study shows that the mouse model absorbs UV-B almost exclusively whereas other animal models such as the rabbit and the guinea pig also contain chromophores that absorb UV-A. The absorptive characteristics of the human lens varies drastically with age. The young lens absorbs primarily UV-A, whereas with age, there are increases in absorptions at 320 nm and out to wavelengths as long as 550 nm. By sectioning human lenses it was found that these changes in absorption properties increased toward the central and the nuclear regions. These absorptive characteristics were then compared to the amount of light reaching the surface of the lens. It was found that UV-B is a minor component of total energy reaching the surface of the human lens and old human lens proteins absorb 2 orders of magnitude more UV-A and visible light than UV-B. It is concluded that it is premature to exclude UV-A or even visible light in the etiology of human cortical cataracts. PMID- 10375443 TI - Volume contents and index PMID- 10375444 TI - Functional MRI of brain activation by eye blinking. AB - Functional magnetic resonance imaging (fMRI) was used to map cortical areas that control eye blinking. T2*-weighted asymmetric spin-echo MRI (or BOLD-blood oxygen level dependent-MRI) was used to detect changes caused by focal variations in blood oxygenation. Six normal volunteers and two cases of dry eye (less than 5-mm Schirmer's test) entered the study. The experimental scheme consisted of three cycles of a two-step sequence: (eyes closed)-(blink or blink inhibition). And to minimize contamination from photic activation, the experiments were carried out in a dark environment and the volunteers reported no light perception during the MR scans. In all eight cases, normal blinking (about one blink every 4 sec) activated areas in the orbitofrontal cortex and in some cases, the visual cortex including the anterior portion of the visual cortex and the primary visual cortex. In severe dry eye, blink-inhibition strongly activated the visual cortex even after irritation due to corneal desiccation was removed by topical anesthesia. The blinking process, especially the rate, appears to be controlled in the orbitofrontal cortex. The significance of visual cortex activation in the dark and in the case of severe dry eye still remains unclear; although it may be associated with attention and arousal. PMID- 10375445 TI - Influence of ophthalmic nerve fibers on choroidal blood flow and myopic eye growth in chicks. AB - Ophthalmic sensory nerve fibers containing substance P and calcitonin gene related peptide' innervate the choroid in mammals and are known to vasodilate choroidal blood vessels. The avian choroid is also innervated by ophthalmic nerve fibers containing substance P and calcitonin gene-related peptide. The present studies were carried out to determine the influence of these sensory fibers on choroidal blood flow in birds and characterize their interaction with manipulations affecting eye growth. In these studies, ChBF was measured using laser Doppler flowmetry in both eyes in the following groups of birds: (1) normal chicks; (2) chicks with right optic nerve transected for 2 weeks; (3) chicks with right optic nerve transected and a goggle over the right eye for 2 weeks; and (4) chicks with right optic and ophthalmic nerves transected and a goggle over the right eye for 2 weeks. The eyes were refracted and various ocular dimensions measured after the blood-flow measurements. It was found that optic nerve transection reduced ChBF to 30% of normal. Placing a goggle (which increases ocular temperature by 4 degrees C) over an optic nerve transected eye nearly doubled choroidal blood flow over that in an optic nerve transected eye without a goggle. Additional transection of the ophthalmic nerve in a goggled optic nerve transected eye, yielded choroidal blood flow that was indistinguishable from that in a nongoggled optic nerve-transected eye. Optic nerve transection had a slight stunting effect on axial growth of the eye. While myopic axial elongation was observed in goggled eyes with the optic nerve cut, the extent of myopia was less than in normal goggled eyes. Ophthalmic nerve transection further reduced the myopia induced by goggling in an optic nerve cut eye. These results suggest that ophthalmic nerve input to the choroid exerts a vasodilatory influence, which is activated in a goggled eye. This increased choroidal blood flow may be in response to elevated ocular temperatures caused by the goggling and this increase appears to be masked in goggled eyes with an intact optic nerve by the reduction in choroidal blood flow normally accompanying myopic eye growth. Our results thus show that the induction of myopic eye growth (as in our optic nerve cut eyes with a goggle) need not be accompanied by a decrease in choroidal blood flow from the baseline no-goggle condition (in this case, with the optic nerve cut). PMID- 10375446 TI - Immunochemical analysis of the sodium channel in rodent and human eye. AB - The presence of the amiloride-sensitive sodium channel (ASSC) in ocular tissues was studied with the aid of a polyclonal antiserum raised against the 14 amino acid peptide QGLGKGDKREEQGL. This sequence corresponds to the region 44-58 of the alpha subunit of the channel, termed ENaC, cloned from rat colon. The antibody titers, measured by the ELISA technique, rose to 1∶2560 4 weeks after immunization, and this bleed was used in all subsequent experiments. Immunoblotting with the polyclonal anti-alphaENaC serum, revealed a major band of 82-86 kDa in extracts prepared from whole bovine or rat retina; a minor component of 92 kDa in the extract from bovine ciliary body may represent a glycosylated species. Immunohistochemistry, using the alphaENaC-specific antiserum, revealed strong fluorescence in specific areas of the rat and human eye. Pronounced labelling was observed in the epithelial cell layer of the retina, the lens, as well as both the pigmented and the nonpigmented epithelium of the ciliary body and the iris. All of the cell layers (epithelium, endothelium and fibroblasts) in the cornea, the blood vessels in the iris, and iris epithelium, were also strongly immunopositive. The somatic body of the photoreceptor cells (cones and rods) in the inner and outer segments could be traced to forming a synapse in both the internal and external portions of the internal nuclear layer. The bipolar cells and ganglia in the neuronal compartment also exhibited occasional immunofluorescence. The method of fixation and the source of the tissue were important parameters for the immunochemical localization of the ENaC. The resolution was very poor when rat eye was fixed in Bouin's solution but this method was satisfactory for human tissues. For rat eye, optimum resolution was obtained with AMeX fixation. This widespread distribution of the ENaC generally colocalizes with the previously observed immunopositivity for the mineralocorticoid receptor such that steroid hormone-mediated ion regulation would appear to add a new parameter to the functional expression of ocular tissues. PMID- 10375447 TI - Phorbol ester modulation of active ion transport across the rabbit conjunctival epithelium. AB - Protein kinase C (PKC) activation elicits diverse cell-type specific effects on key epithelial transporters. The present work examined the influence of phorbol esters, which are known activators of PKC isoenzymes, on the short-circuit current (Isc), a direct measure of net transcellular electrolyte transport, of the rabbit conjunctiva. In this preparation, the Iscmeasures a Na+-dependent, bumetanide-inhibitable Cl-transport in the basolateral-to-apical direction plus an amiloride-resistant Na+absorptive process in the opposite direction. Additions of phorbol 12-myristate-13-acetate (PMA) to the basolateral bathing media did not affect the transepithelial electrical parameters; but its introduction to the apical bath at 1 and 10 micrometers elicited a transient ( approximately 2 min duration) Iscspike followed by a sustained reduction relative to the control level. Such PMA-elicited Iscreductions were from 14. 0+/-2.0 to 3.1+/-0.8 microA cm-2(+/-s.e.m.'s, n =3) at 1 micrometer and from 16.5+/-1.9 to 4.6+/-0.7 microA cm-2(n =22) at 10 micrometers. The former concentration failed to produce extensive Iscreductions in 3 other experiments. Similar results were obtained with phorbol 12, 13-dibutyrate (PDBu). Its apical administration at 0.1 micrometer reduced the Iscfrom 18.5+/-4.1 to 7.8+/-2.0 (n =3), and from 16. 5+/ 2.9 to 6.9+/-1.2 (n =7) when introduced at 1 micrometer. The phorbol-evoked Iscreductions occurred without a simultaneous change in transepithelial resistance (Rt). However, after about 15-20 min, Rtgradually declined by about 25%. In contrast to these results, treatment with a phorbol ester known not to activate PKC (4-alpha-PMA) did not affect the electrical parameters when added at 10 micrometers. PMA- and PDBu-evoked Iscreductions could be obtained with conjunctiva that were (1) pretreated with bumetanide, (2) bathed in Cl--free media, and (3) pretreated with amphotericin B, changes consistent with a likely inhibition of the basolateral Na+/K+pump. Such Iscinhibitions were attenuated with conjunctiva pre-exposed to 1 micrometer staurosporine, a nonselective kinase inhibitor known to suppress PKC activity. Staurosporine, in itself, produced a rapid 26% Iscinhibition (n =15) along with a 17% Rtincrease upon its apical introduction. These electrical responses were less extensive in Cl--free media and absent in amphotericin B-treated conjunctiva, suggesting the presence of a kinase-mediated regulation of apical channels for both Na+and Cl-. Overall, these results imply that in addition to previously demonstrated epinephrine-elicited, up-regulation of Cl-secretion, mechanisms may also exist, via PKC activation, to suppress Na+/K+pumping and consequently reduce transepithelial transport rates. PMID- 10375448 TI - Gap junctions containing alpha8-connexin (MP70) in the adult mammalian lens epithelium suggests a re-evaluation of its role in the lens. AB - A missense mutation in one of the three lens connexins, alpha8-connexin, has been recently shown to be the genetic basis of the zonular pulverant lens cataract. This connexin had been considered to be expressed only in lens fibre cells. The present studies show that alpha8-connexin is also expressed in the lens epithelial cell layer. For this study, the distribution of gap junctions in the adult bovine lens has been investigated by confocal immunofluorescence microscopy using antibodies against alpha8-connexin (MP70) and alpha1-connexin (Cx43). In addition to the anticipated localisation of alpha8-connexin to the broad faces of lens fibre cells as reported in other species, alpha8-connexin was also found colocalized with alpha1-connexin at plaques in the lateral epithelial-epithelial plasma membranes of the bovine lens. These data suggest that mixed alpha8 connexin/alpha1-connexin plaques are between epithelial cells at their apico lateral plasma membranes, rather than between epithelial and fibre cells. Indeed, freeze fracture analyses of the epithelial-fibre cell interface failed to reveal gap junctions connecting the epithelium and the underlying fibre cells. Importantly, microdissection and subsequent immunoblotting of lens epithelium samples confirmed the immunolocalisation results. The data suggest mature mammalian lens epithelial cells could form either heteromeric, heterotypic and/or mixed homomeric-homotypic gap junctional complexes with unique physiological properties, an important point when considering the role of epithelial cell connexins in cataractogenesis. PMID- 10375449 TI - Characterization of a novel C-kinesin (KIFC3) abundantly expressed in vertebrate retina and RPE. AB - Many forms of intracellular transport are mediated by microtubule-dependent motors of the kinesin superfamily (KIFs). To identify kinesins expressed in human retina and RPE, we used degenerate primer RT-PCR to amplify a approximately 440 bp kinesin motor domain fragment from human retinal and RPE messenger RNAs. Four distinct kinesins were detected: one C-kinesin (HsKIFC3); one kinesin from the unc104/KIF1 family [HsKIF1A]; and the ubiquitous and neuronal forms of conventional kinesin heavy chain [HsuKHC and HsnKHC]. The C-kinesin HsKIFC3 comprised 33.3% of the retinal clones and was 60% identical to FKIF2, the most abundant kinesin detected in a previous screen of fish retina and 95% identical to a fragment of MmKifC3 recently amplified from mouse brain. Elsewhere we have reported the sequence of HsKIFC3 and shown that it maps to the same locus on chromosome 16q13-q21 as Bardet-Biedl syndrome Type II, a hereditary retinal degeneration. We describe here the kinesin PCR screen of human retina and RPE and examine the tissue and subcellular distribution of KIFC3 in both fish and human retina using an antibody raised against a peptide conserved between FKIF2 and HsKIFC3. This peptide antibody identified a single approximately 80 kDa band in Western blots of fish and human retina and RPE. In both fish and human retina this antibody strongly labeled photoreceptor terminals in the outer plexiform layer, suggesting that FKIF2/KIFC3 may play some role in the photoreceptor synapse. PMID- 10375450 TI - Establishment of T-helper type 2 clone-induced eosinophilia in mouse conjunctiva. AB - Eosinophil infiltration is one of the aspects of the inflammatory response to an allergen. The aim of this study was to establish T-helper type-2 (Th2)-induced conjunctival eosinophilia in mice to evaluate the effect of anti-allergic drugs. Th2 clone, D10.G4.1. (D10) with its specific antigen, conalbumin was co-injected into conjunctival tissue in mice. At 3-6 hr after D10 injection, eosinophil and neutrophil recruitment into conjunctival tissue was observed. Cellular infiltration into conjunctival tissue reached a maximum level between 24-48 hr after D10 injection. The number of eosinophils which infiltrated after 24 hr was dose dependent on T cell injection titer from 1x10(4), 1x10(5)to 1x10(6)cells site-1. The ratio of eosinophils to neutrophils at 24 hr after D10 cell (1x10(5)cells site-1) injection was about 9 to 1. The eosinophil infiltration into conjunctival tissue was significantly reduced by the intraperitoneal administration of anti-IL-5 monoclonal antibody (100 micrograms animal-1). Anti IL-4 monoclonal antibody (500 micrograms animal-1) partially inhibited eosinophilia. The combined inhibitory effect of anti-IL-4 and anti-IL-5 monoclonal antibodies was larger than the inhibitory effect of anti-IL-5 monoclonal antibody alone. In conclusion, a Th2-induced mouse conjunctival eosinophilia model was established in which D10 activation results in IL-4 and IL 5 release. These cytokines elicit eosiniophil infiltration. This response shows that the model will be effective for screening candidates of anti-allergic drugs. PMID- 10375451 TI - Monoclonal IgA antibodies protect against Acanthamoeba keratitis. AB - Acanthamoeba keratitis is a rare, yet sight-threatening corneal infection. Ocular infection does not appear to induce protective immunity as repeated corneal infections occur in both humans and experimental animals. However, we have recently demonstrated that activation of the common mucosal immune system by oral immunization with Acanthamoeba antigens protects both Chinese hamsters and pigs against ocular infection with A. castellanii. Protection correlates closely with the appearance of anti- Acanthamoeba antibodies in the tears. To test the hypothesis that oral immunization induces specific protective IgA antibodies, two monoclonal IgA antibodies specific for Acanthamoeba antigens were generated. Both antibodies detected epitopes on the surface of fixed Acanthamoeba trophozoites. When delivered intraperitoneally, one monoclonal antibody (14E4) was detected in stool and tear samples. This clone also protected naive animals against ocular challenge with Acanthamoeba trophozoites (43% infection rate compared to a 91% infection rate in animals receiving control IgA). In vitro functional studies showed that neither antibody induced encystment or directly killed Acanthamoeba trophozoites. However, both monoclonal anti- Acanthamoeba IgA antibodies produced a three-fold inhibition in the adherence of trophozoites to corneal epithelial cells in vitro. These data show that monoclonal anti- Acanthamoeba IgA antibodies can protect against Acanthamoeba keratitis and suggest that this occurs by inhibiting adhesion of the parasite to the corneal epithelium. PMID- 10375452 TI - Effects of K+Channel blockers on acetylcholine-induced vasodilation in guinea-pig choroid. AB - The purpose of this study is to clarify which K+channels contribute to the acetylcholine (ACh)-induced vasodilation from the diameter changes in arterioles of the guinea-pig choroid. The choroid was isolated from the guinea-pig eyeball, pinned flat on a silicone rubber plate and superfused with warmed oxygenated (35 degrees C) Krebs solution. Diameters of choroidal arterioles were measured using video microscopy and a computer program for analysis. The effects of K+channel inhibitors (glibenclamide, tetraethylammonium [TEA], apamin and charybdotoxin [ChTX]) on the ACh-induced vasodilation were examined in arterioles which had been constricted by either norepinephrine (NE) or high K+solution. In NE (10( 5)m)-constricted arterioles, the combination of nitroarginine (10(-4)m) and indomethacin (10(-5)m) reduced ACh (10(-6)m)-induced vasodilatation by 24%. When high K+solution was used to constrict the arterioles, ACh-induced vasodilation was abolished by nitroarginine and indomethacin. In the presence of nitroarginine and indomethacin, the ACh-induced dilatation of NE-constricted arterioles was attenuated by TEA (10(-3)m), apamin (10(-7)m), and ChTX (10(-7)m) but not by glibenclamide (2x10(-5)m). Simultaneous application of apamin and ChTX inhibited the ACh (10(-6)m)-induced dilatation by 85%. In arterioles of guinea pig-choroid, nitric oxide and prostacyclin are not main mediators in ACh-induced vasodilation. Simultaneous activation of a set of Ca2+-sensitive K+channels may take most part of ACh-induced vasodilation. PMID- 10375453 TI - Changes of GABA metabolic enzymes in acute retinal ischemia. AB - It is reported that GABA accumulates in Muller cells in ischemic and diabetic rat retina. To investigate the mechanism of GABA accumulation in Muller cells, we localized GABA and glutamate in ischemic rat retina and measured the activity of GAD and GABA-T, enzymes involved in GABA metabolism. Using general anesthesia, we incised the bulbar conjunctiva of the rat around the limbus and clamped the left optic nerve. A sham operation was performed on the right eyes. Ocular ischemia was sustained for 30, 60 and 90 minutes. Rat eyes were enucleated immediately after ischemia and prepared for immunohistochemistry and enzyme activity measurement. Glutamate-like immunoreactivity (Glu-IR) in the sham-operated rat retina was observed in all retinal layers, showing intense staining in the nerve fiber layer (NFL), ganglion cell layer (GCL), and inner plexiform layer (IPL). Glu-IR increased in the outer plexiform layer (OPL) and outer nuclear layer (ONL) in an ischemic time-dependent manner. GABA-like immunoreactivity (GABA-IR) in sham-operated rat retina was observed in NFL, GCL, IPL and inner nuclear layer (INL). When the ischemic time was extended, GABA-IR intensely stained Muller cells. GAD activity was not changed in ischemic rat retina as compared to normal rat retina, but GABA-T activity was significantly decreased in ischemic rat retina. These results suggested that glutamate was induced by ischemia and was converted to GABA by GAD activity. Increased GABA was not metabolized because GABA-T activity was decreased. GABA accumulation in Muller cells progressed during the change in activity of these metabolic enzymes. PMID- 10375454 TI - Maintenance of retinoid metabolism in human retinal pigment epithelium cell culture. AB - If transplantation of cultured retinal pigment epithelium (RPE) or iris pigment epithelium (IPE) is to be successful in the treatment of ocular disease, it is imperative to demonstrate that these cells can perform all of their necessary metabolic functions. Unfortunately, a critical function of the RPE, retinoid metabolism, is often lost rapidly in culture. We have examined whether or not nonspecific proteolytic enzymes commonly used in cell isolation and serial passaging may be responsible for this loss of function, and we have investigated novel isolation and passaging techniques which can alleviate this loss of retinoid metabolism.RPE cells were obtained from human donor eyes by enzymatic and nonenzymatic methods. Cells were cultured either on control tissue culture inserts or on inserts coated with a layer of thermally responsive poly(N isopropylacrylamide-co-cinnamoylcarbamidemethylstyrene). Upon confluence, cells were detached either by trypsinization or by lowering dish temperature. Retinoid metabolism of cells was assessed after isolation and culture by incubating membrane fractions with3H-all- trans -retinol. Retinoid metabolism was also measured in freshly isolated IPE, corneal endothelium (CE), an RPE cell line (D407), and two hepatocyte cell lines (Hepa 6 and HepG2). Membrane fractions from cells isolated nonenzymatically or using collagenase/hyaluronidase formed 11- cis -retinol, retinal isomers and retinyl esters. Retinoid metabolism of RPE cells freshly isolated by trypsinization showed no 11- cis -retinal and little 11- cis retinol formation. Nondamaged cells cultured on thermally responsive surfaces detached in sheets upon temperature change. They showed metabolism similar to that of cells freshly isolated by nonenzymatic means. After trypsinization, confluent cultures dissociated into individual cells, but these cells showed poor retinoid metabolism, including no detectable retinyl esters or 11- cis -retinoid isomers. IPE, CE and Hepa 6 did not show any retinoid metabolism. D407 and HepG2 produced retinals, but not the 11- cis isomer.RPE cells isolated using trypsin lose the ability to form critical intermediates in the visual cycle. Collagenase/hyaluronidase or nonenzymatic cell isolation techniques enable these functions to be maintained. After cell culture, thermally responsive surfaces allow nonenzymatic cell detachment and excellent maintenance of retinoid metabolism. PMID- 10375455 TI - Acetyl- L -carnitine decreases glycation of lens proteins: in vitro studies. AB - Although the role of carnitine system in the ocular tissues is not clearly understood, earlier studies showed that lenticular levels of L -carnitine were the highest among ocular tissues and there was a dramatic depletion of lenticular L -carnitine and acetyl- L -carnitine in streptozotocin-diabetic rats. As protein glycation has been implicated in the development of several diabetic complications including cataracts, this study was initiated to show the possible effects of L -carnitine and acetyl- L -carnitine on the glycation and advanced glycation (AGEs) of lens proteins. Calf lens soluble fraction (crystallins) was incubated with 50 m m glucose (containing14C glucose) with or without 5-50 m ml carnitine, 5-50 m m acetyl- L -carnitine and 5-50 m m acetyl salicylic acid, for 15 days. The results show that while L -carnitine did not have any effect on in vitro glycation of lens crystallins, acetyl- L -carnitine and acetyl salicylic acid decreased crystallin glycation by 42% and 63%, respectively-this decrease was concentration dependent. Glycated crystallins were separated on HPLC which showed that the rate of glycation is in the following order: alpha>beta>gamma. Interestingly, acetyl- L -carnitine inhibited glycation of alpha crystallin more than other crystallins. In vitro incubations with [3H-acetyl] acetyl- L carnitine showed that acetyl- L -carnitine acetylates lens crystallins (non enzymatically) and alpha crystallin is the major acetylated protein. Furthermore, there was a 70% reduction in anti-AGE antibody reactivity when 50 m m acetyl- L carnitine was included in the incubation of lens crystallins and 10 m m erythrose, suggesting that inhibition of glycation by acetyl- L -carnitine also affected the generation of AGEs. This in vitro study shows, for the first time, that acetyl- L -carnitine could acetylate potential glycation sites of lens crystallins, and protect them from glycation-mediated protein damage. PMID- 10375456 TI - Effects of antimetabolite induced cellular growth arrest on fibroblast-fibroblast interactions. AB - The use of single five-minute applications of antimetabolites during glaucoma filtration surgery has significantly reduced the occurrence of post-operative scarring and bleb failure. However, surgery for some patients is still unsuccessful, despite the use of antiproliferative agents, due to formation of scar tissue at the drainage site. It is not known if cells growth arrested in the treated area with a single application of antimetabolites influence the activity of adjacent non-treated cells. We hypothesise that the activity of non-treated cells recruited to the wound site may be involved in post-operative scarring. The aim of this study was to investigate the effects of antimetabolite induced cellular growth arrest on cell-cell interactions using in vitro techniques.Tenon's capsule fibroblast cultures were growth arrested by exposure for 5 minutes to mitomycin-C (0.001, 0.01 and 0.1 mg ml-1), 5-fluorouracil (0.25, 2.5 and 25 mg ml-1) or phosphate buffered saline (PBS). Following a period of serum-starvation, conditioned media (CM) were subsequently collected from the cells at intervals up to 29 days post-treatment. Correction for cell number was made prior to supplementation of serum-free medium with CM. CM were assessed for ability to support or inhibit normal non-treated fibroblast proliferation, migration and collagen contraction. Conditioned media collected from cells growth arrested with MMC or 5FU stimulated normal fibroblast proliferation, migration and collagen contraction in excess of non-conditioned serum-free medium. Peaks of fibroblast activity in CM differed according to which drug and concentration had originally been given to the treated cells. This study has demonstrated that CM collected from fibroblasts treated for 5 minutes with a range of concentrations of antimetabolites can differentially influence normal non-treated fibroblast activity. This in vitro data suggests that despite entering growth arrest, fibroblasts may still influence the behaviour of other cells via soluble mediators. They may have implications in the clinical setting, in that it may not be sufficient to suppress proliferation alone to prevent fibroblast behaviour associated with scarring after glaucoma filtration surgery. PMID- 10375457 TI - Differential inhibition of three peptidase activities of the proteasome in human lens epithelium by heat and oxidation. AB - The proteasome is a large protease complex that is thought to be responsible for proteolytic removal of damaged proteins. We have previously shown that the level of proteolytic activity due to the proteasome is lower in lens epithelium from human cataractous lenses compared to the activity in epithelium from clear donor lenses. This study aimed to characterize the three main peptidase activities of the proteasome in human lens epithelium with respect to kinetic properties and sensitivity to heat and oxidation. Human lens epithelia were obtained from cataract surgery and analysis performed on pools of epithelial cell cytoplasm. Using the fluorogenic peptide substrates Suc-Leu-Leu-Val-Tyr-AMC (LLVY), Boc-Val Gly-Arg-AMC (VGR) and Z-Leu-Leu-Glu-betaNA (LLE), Km-values of 56, 678 and 108 micrometers were obtained. All peptidase activities were inhibited by lactacystin, a specific proteasome inhibitor, but at very different rates; with LLVY-hydrolysing activity being the most sensitive (Ki50%=0.15 micrometers). Thermostability was investigated by performing the proteolytic assay at 20 degrees, 37 degrees and 53 degrees C. The trypsin-like activity, as measured by VGR, was completely stable at 53 degrees C for at least 24 hr whereas hydrolysis of LLVY and LLE declined after a few hours at 37 degrees C. Oxidative inhibition was induced by incubation of the samples in 0.5 m m H2O2for 1 or 24 hr. One hour exposure to H2O2caused moderate inhibition of all peptidase activities. The activity could be partially restored by adding 1 m m dithiotreitol, indicating the dependency on intact SH-groups. After 24 hr, peptidase activities were decreased to 25% (LLVY), 73% (VGR) and 44% (LLE) of corresponding control. This inhibition was irreversible for VGR and LLE, but could be partly prevented by the presence of heat shock protein 90 (LLVY and VGR) or alpha-crystallin (LLVY). These data show that the peptidase activities of the human lens proteasome can be modulated by metabolites, such as reactive oxygen species, and by endogenous proteins such as alpha-crystallin and heat shock protein 90. PMID- 10375458 TI - Antibodies to lens epithelium-derived growth factor (LEDGF) kill epithelial cells of whole lenses in organ culture. PMID- 10375459 TI - Evolution and physiology of the corticotropin-releasing factor (CRF) family of neuropeptides in vertebrates. AB - Corticotropin-releasing factor (CRF), urotensin-I, urocortin and sauvagine belong to a family of related neuropeptides found throughout chordate taxa and likely stem from an ancestral peptide precursor early in metazoan ancestry. In vertebrates, current evidence suggests that CRF on one hand, and urotensin-I, urocortin and sauvagine, on the other, form paralogous lineages. Urocortin and sauvagine appear to represent tetrapod orthologues of fish urotensin-I. Sauvagine's unique structure may reflect the distinctly derived evolutionary history of the anura and the amphibia in general. The physiological actions of these peptides are mediated by at least two receptor subtypes and a soluble binding protein. Although the earliest functions of these peptides may have been associated with osmoregulation and diuresis, a constellation of physiological effects associated with stress and anxiety, vasoregulation, thermoregulation, growth and metabolism, metamorphosis and reproduction have been identified in various vertebrate species. The elaboration of neural circuitry for each of the two paralogous neuropeptide systems appears to have followed distinct pathways in the actinopterygian and sarcopterygian lineages of vertebrates. A comparision of the functional differences between these two lineages predicts additional functions of these peptides. PMID- 10375460 TI - Fertility decline in aging roosters is related to increased testicular and plasma levels of estradiol. AB - The relationships between testicular and plasma hormone levels and the decline in fertility in aging roosters were examined. Body mass, testicular mass, and fertility were measured in roosters from 20 to 72 weeks of age. Plasma was assayed for LH and testosterone, and estradiol and testicular extracts were assayed for testosterone and estradiol contents. Fertility increased rapidly in young roosters to a peak of 96.2 +/- 3.9% at 37 weeks of age. Thereafter, fertility declined and by 72 weeks of age was significantly lower than at 37 weeks. Plasma LH reached 16.8 +/- 2.5 ng/ml at 27 weeks and remained high until 60 weeks of age, when it decreased significantly. Plasma and testicular testosterone levels increased from low levels in young birds to a peak that coincided with highest fertility and declined thereafter. Plasma and testicular estradiol showed a striking inverse relationship with testosterone. Plasma estradiol was 29.4 +/- 4.0 pg/ml in 20-week-old birds, decreased rapidly as testosterone increased, and increased again in older birds as testosterone decreased. Thus, the decline in fertility in aging roosters was associated with a decrease in plasma LH and testosterone and an increase in plasma and testicular estradiol. It is suggested that plasma levels of LH and testosterone in roosters are regulated by a negative feedback mechanism involving estradiol that is produced not only by the aromatization of testosterone in the brain but also by peripheral estradiol originating in the testes and that estradiol has a major role in the decline in fertility in aging roosters. PMID- 10375461 TI - Hormone coexpression in the adult toad endocrine pancreas: double-label immunofluorescence under basal conditions and after glucose injection. AB - We have investigated the type and frequency of hormone coexpression in the endocrine pancreas of amphibians both under basal conditions and after sustained glucose loading. Adult male specimens of the wild toad Bufo arenarum were injected with a 50% (w/v) glucose solution (2 g/100 g) for 2 days, while control animals received an equal volume of the vehicle. Serum glucose levels were measured at the time of sacrifice and the pancreatic free lobe was processed for light microscopy. A double-labeling immunofluorescence study was performed for the detection of insulin (I), glucagon (G), somatostatin (S), and pancreatic polypeptide (PP). Heterospecific antisera against the following hormone combinations were used for their detection and immunocytochemical localization: I+G, I+PP, G+PP, S+G, and S+PP; visualization of the reacted IgG's was effected by fluorescein- and rhodamine-conjugated species-specific antibodies as fluorophores. Intracellular hormone coexpression was found to occur in the combinations G+PP, S+G, and S+PP. Moreover, glucose administration caused, together with a marked hyperglycemia (123 +/- 17 vs 23 +/- 1 mg/dl; P < 0.05), a decrease in the fraction of cells containing both G and PP together (from 106.3 +/- 8.1 to 26 +/- 4 cell/mm2) along with a reciprocal rise in the number of cells possessing G alone (from 128.7-152.3 to 235.9-274 cell/mm2). The fewer number of cells coexpressing either of the other two hormone combinations, however, were unaffected by glucose injection. With respect to the simultaneous measurement of I+G and I+PP, no cells were detected with both hormones of either pair, and the I containing cells were more frequent in each instance in the control toads (264.8 +/- 22.3 to 269.2 +/- 27 cell/mm2). For both combinations, however, this value diminished significantly in the glucose-treated animals (108 +/- 2 cell/mm2 for I+G and 112.1 +/- 7. 8 cell/mm2 for I+PP). While the G-containing cells became more numerous (rising to 235.9 +/- 12.4, 274 +/- 26, and 250.4 +/- 23.7 cell/mm2 for I-G, G-PP, and G-S combinations, respectively), the PP- and S-containing cells remained unaffected. We conclude that the copresence of different hormones within the same cell is a relatively common finding in the non-I-secreting elements of the adult toad pancreas and that the proportions of specific cell types are affected by glucose administration. We thus propose that intracellular hormonal coexpression in this fashion may well represent a rapid and efficient regulatory mechanism for compensating for the metabolic stress imposed by glucose loading. PMID- 10375462 TI - A newly established ELISA showing the effect of the androgenic gland on secondary vitellogenic-specific protein in the hemolymph of the crayfish Cherax quadricarinatus. AB - A quantitative enzyme-linked immunosorbent assay (ELISA) was developed to monitor the onset of secondary vitellogenesis in Cherax quadricarinatus females and in intersex individuals (having both male and female reproductive systems) after removal of the androgenic gland (AG). As a prerequisite for the assay, the 106 kDa polypeptide was separated from newly laid C. quadricarinatus eggs by SDS PAGE, and anti-106-kDa antibody was raised in rabbit. The specificity of the anti 106-kDa polypeptide for proteins specific for the hemolymph of secondary vitellogenic females was confirmed by double immunodiffusion and immunoblot cross reactivity tests. A characteristic standard ELISA curve, using egg high-density lipoprotein (HDL), showed linearity between 16 and 500 ng (r = 0. 953) and was sensitive for amounts as low as 8 ng. The inter- and intraassay coefficients of variance were 14.8 and 7.2%, respectively. Only traces of egg HDL equivalents were detected in the hemolymph of male and primary-vitellogenic females (11 to 110 microg/ml), confirming the specificity of the assay, whereas high levels of such a protein (8-35 mg/ml) were detected in the hemolymph of secondary vitellogenic females. Removal of the AG from intersex individuals leads to a significant increase in the concentration of vitellogenic-specific protein in the hemolymph (up to 2 mg/ml). Moreover, a significantly lower concentration was found in females subjected to AG transplant (79.3 microg/ml). The ELISA thus provided an accurate and sensitive tool to investigate the influence of the AG on the expression of a vitellogenic-specific protein in female and intersex C. quadricarinatus, confirming the central role of this gland in tuning sexual plasticity in this species. PMID- 10375463 TI - Effects of prolactin and growth hormone on plasma immunoglobulin M levels of hypophysectomized rainbow trout, Oncorhynchus mykiss. AB - Immunoglobulin M (IgM) is a major component of the humoral immune system of teleosts. This study examines the effects of hypophysectomy and subsequent replacement with prolactin (PRL) or growth hormone (GH) upon the plasma IgM levels of the rainbow trout (Oncorhynchus mykiss). Plasma IgM levels of the hypophysectomized fish were decreased to 30% of those in sham-operated fish 1 or 4 weeks after operation. Implantation of a cholesterol pellet containing salmon PRL or GH restored plasma IgM levels of the hypophysectomized fish, suggesting important roles for PRL and GH in the regulation of circulating IgM level in trout. PMID- 10375464 TI - Reproductive condition and behavior in relation to plasma levels of gonadal steroids in the spiny damselfish Acanthochromis polyacanthus. AB - Gonadal condition and plasma levels of gonadal steroids were measured in relation to behavior in the biparental brood-protecting spiny damselfish Acanthochromis polyacanthus. Fish were captured by SCUBA divers from natural populations on Australia's Great Barrier Reef and immediately bled underwater, following diver or video observation of precapture behaviour. In winter (July), most fish were nonterritorial, with a low proportion of mature males, or vitellogenic females present. In spring (November), most fish were territorial with broods of young at varying stages of maturity, and all stages of gonadal development were represented. Territorial males were larger than nonterritorial fish, but territorial fish that had large (older) young (the end of the brooding phase) had lower condition factors than fish at other stages. Males of all gonad stages had a high proportion of spermatozoa in the testis, but this was higher in November than in July. Ovaries of females commonly had several classes of developing follicles present, although fish that were brooding large young had regressed ovaries with a high incidence of atresia. Plasma levels of testosterone (T) and 11-ketotestosterone were elevated in males of advanced gonadal maturity, and also in relation to recent or imminent spawning behaviour, but there were no changes in plasma 17, 20beta-dihydroxy-4-pregnen-3-one (17,20betaP), which was near assay detection limits at all times. Females had elevated T and 17beta-estradiol (E2) in association with vitellogenesis and elevated T in relation to spawning activity in some fish, but as in males, 17,20betaP levels were low and unchanging. Territorial females without young had lower cortisol levels than nonterritorial fish, or females protecting young. The results confirm the importance of elevated androgens to spawning activity in territorial male fish, but not females where endocrine activity is more closely related to stage of ovarian development. Extended brooding appears to inhibit vitellogenesis, perhaps via a stress-related mechanism. PMID- 10375465 TI - Effects of photoperiod on salmon GnRH mRNA levels in brain of castrated underyearling precocious male masu salmon. AB - Previous studies have suggested that activation of salmon gonadotropin-releasing hormone (sGnRH)-producing neurons is induced by the combined effects of photoperiod and steroid hormones in underyearling males of the masu salmon, Oncorhynchus masou. The present study further assesses the effects of photoperiod and steroid hormones on sGnRH synthetic activity and examines the changes in sGnRH mRNA levels in the brains of castrated underyearling precocious male masu salmon by manipulating the photoperiod for 60 days from August through October. In castrated males in which plasma testosterone levels decreased to low levels, sGnRH mRNA levels in the preoptic area (POA) increased under a short photoperiod (8L-16D), whereas they remained at low levels under a long photoperiod (16L-8D) for a 2-month duration. In sham-operated males, sGnRH mRNA levels in the ventral telencephalon and those in the POA increased in October with testicular maturation even under a long photoperiod with a delay of 1 month compared with the short photoperiod group. These results suggest that preoptic sGnRH-producing neurons receive short photoperiodic signals and that either short photoperiod or steroid hormone secretion is required for the activation of sGnRH synthesis in underyearling precocious male masu salmon. PMID- 10375467 TI - Plasma steroid interactions during high-density green turtle nesting and associated disturbance. AB - Raine Island in the Northern Great Barrier Reef constitutes an extremely high density green turtle (Chelonia mydas) rookery. On this island, competitive interactions for nesting space and subsequent disturbance of individual nesting are widespread. High-density nesting often delays successful oviposition by one or more nights. There is little information on how hormones in female reptiles interact during competitive reproductive events in such high-density nesting populations. In this three-part study we investigated the interactions between density (within and between rookery/ies), nesting success and failure, and plasma steroid profiles in green turtles. First, we compared levels of plasma corticosterone (B) and combined testosterone + 5alpha-dihydrotestosterone (T + DHT) in turtles during five stages of oviposition in both a high-nesting-density sector (1 turtle/m2) and a low-nesting-density sector (0.1 turtle/m2). Second, we investigated the relationship between increasing delays (0, 1, 2, 3, and 6 days) in successful oviposition and the plasma steroids B and T + DHT. Third, we assessed a comparative measure of steroid hormone levels of females at low density sites on Raine Island (high-density rookery) and Number Seven Sandbank (low-density rookery). Despite a significant trend suggesting high-density nesting turtles elaborated more plasma B than turtles in low-density sectors, the magnitude of this increase was small. We suggest that this increase may be an artifact of increased metabolic demand and hence catabolism of energy substrates associated with high-density nesting. Plasma T + DHT remained stable in response to density-dependent effects associated with nesting. Furthermore, prolonging successful oviposition because of multiple nightly disturbance failed to elicit any change in either plasma B or T + DHT. These data suggest that green turtles may be exhibiting adrenal desensitization to prevent both physical and behavioral disturbances interfering with reproduction. We suspect that down-regulating the acute adrenocortical response may represent an adaptive trade-off mechanism for optimizing current reproductive success at the potential expense of survivorship. PMID- 10375466 TI - Patterns of urinary and fecal steroid excretion during the ovarian cycle and pregnancy in the African elephant (Loxodonta africana). AB - The aims of the present study were to (i) determine the relative abundance of the 5alpha-reduced progestins 5alpha-pregnane-3-ol-20-one (5alpha-P-3OH) and 5alpha dihydroprogesterone (5alpha-DHP) and progesterone (P4) in African elephant feces and to establish improved fecal progestin assays for monitoring ovarian function; and (ii) describe longitudinal profiles of urinary and fecal progestin and estrogen metabolites during pregnancy. Matched urine and fecal samples were collected weekly from six adult females throughout 18 nonfertile cycles and two complete pregnancies (89 and 93 weeks duration). Fecal samples were lyophilized and extracted with 80% methanol in water and immunoreactive 5alpha-P-3OH, 5alpha DHP, and P4 and (for pregnant females only) estrone (E1) and estradiol (E2) determined by enzyme immunoassay. Urine samples were hydrolyzed, ether-extracted, and assayed for 5alpha-P-3OH, E1, and E2. HPLC cochromatography of fecal extracts with various radioactive progestin tracers confirmed the presence of large amounts of both 5-reduced progestins (5alpha-P-3OH > 5alpha-DHP) but not of P4. 5 Reduced progestins (but not P4) were excreted in a cyclic pattern and levels were significantly correlated with urinary 5alpha-P-3OH. Fecal 5alpha-P-3OH showed the more pronounced and consistent luteal-phase elevation and a better correspondence to urine with respect to timing of the luteal-phase rise. Fecal and urinary 5 reduced progestins increased gradually during early pregnancy to maximum values around week 40-45. Levels gradually declined during the second half of pregnancy, reaching baseline values 2 days before parturition. Urinary estrogens did not show any cyclic pattern during the preconception period and levels remained low during the first 30 weeks of gestation. Thereafter, there was a rapid 10- to 20 fold increase to maximum values at mid-pregnancy, followed by a gradual decline to birth. There was no mid-pregnancy elevation in fecal estrogens, but there was a modest increase in E1 during the second half of gestation. PMID- 10375468 TI - Enhancement by proopiomelanocortin-derived peptides of growth hormone and prolactin secretion by bullfrog pituitary cells. AB - Corticotrophs in the bullfrog (Rana catesbeiana) are situated mainly in the rostral region of the anterior lobe of the pituitary gland, which receives its blood supply primarily from the portal vessel. On the assumption that the proopiomelanocortin (POMC)-derived peptides released into the pituitary circulation may influence the function of other pituitary cells situated downstream, the effects of three POMC-derived peptides, namely, N-terminal peptide of POMC (NPP), adrenocorticotropic hormone (ACTH), and joining peptide (JP), on the secretion of growth hormone (GH) and prolactin (PRL) by bullfrog dispersed anterior pituitary cells were examined. NPP and ACTH, but not JP, stimulated the release of GH and PRL in a concentration-dependent manner. It was also found that ACTH1-17, but not alpha-melanocyte-stimulating hormone, was effective in enhancing GH and PRL release. A marked difference between the response to NPP and ACTH and the response to thyrotropin-releasing hormone employed as a reference secretagogue in terms of the time required for stimulating the release of GH and PRL was noted. Northern blot analysis of GH and PRL mRNA levels and radioimmunoassay for GH and PRL in the cultured cells revealed that ACTH increases the syntheses of both pituitary hormones as well. The possibility that NPP and ACTH act on neighboring cells to maintain their overall secretory function is discussed. PMID- 10375469 TI - Low fertility in aging roosters is related to a high plasma concentration of insulin and low testicular contents of ACTH and lactate. AB - Fertility in roosters peaks between 30 and 40 weeks of age and declines rapidly from about 50 weeks of age. Low-fertility, aging roosters have a higher density of elongated spermatids attached to Sertoli cells than do high-fertility roosters, but display normal spermatogenesis and ejaculated spermatozoa. Plasma levels of insulin and lactate and testicular contents of ACTH and lactate were compared in Cornish roosters aged 27 weeks (early state of sexual maturity), 37 weeks (high fertility), 67 weeks (reduced fertility), and 72 weeks (low fertility). Insulin may act as an endocrine regulator of Sertoli cell function, and ACTH as an autocrine regulator of Leydig cells for androgen production and as a paracrine regulator of Sertoli cells by amplifying FSH response. Lactate is the primary energy substrate of spermatocytes and spermatids in the adluminal compartment. Roosters aged 67 and 72 weeks had higher (P < 0.05) plasma insulin levels but lower (P < 0.05) testicular lactate content than roosters aged 27 and 37 weeks. The lower lactate content in testes of low-fertility roosters may reflect an increased consumption of lactate due to the higher density of elongated spermatids. Furthermore, the content of testicular ACTH was lower in low-fertility roosters than in 27-week-old roosters. These results suggest that ACTH may be involved indirectly in the mechanism responsible for the high density of spermatids in the tubuli and the lower spermatozoa concentration in the ejaculate of low-fertility roosters, as was reported in previous studies, since this hormone may serve as a paracrine regulator of Sertoli cell function. PMID- 10375470 TI - Fasting increases the plasma glucagon response in the migratory garden warbler (Sylvia borin). AB - Acute pancreatic hormonal responses to oral glucose loads were investigated in garden warblers during the prolonged fast that follows their autumnal migratory fattening. Plasma glucose, free fatty acids, beta-hydroxybutyrate, insulin (INS), and glucagon (GLN) were measured prior to and 10 min after an oral glucose load in three groups of birds: One had food ad libitum, and the other two were either food restricted or food deprived down to a given (low) body mass level. Ten minutes after the glucose load, plasma glucose levels increased significantly in all three groups (range of mean values: basal, 15.1-16.0; glucose-stimulated, 19.1-23.7 mmol/L). A smaller increase in food-restricted/deprived groups was not statistically significant. Free fatty acid levels (1.0-1.5 mmol/L) after 10 min were unchanged, while beta-hydroxybutyrate decreased to similar levels in food restricted/deprived and control groups (basal, 3.3-4.2; glucose-stimulated, 1.9 2.4 mmol/L). Insulin increased and glucagon decreased in response to oral glucose loads. However, initial levels and responses of plasma insulin to glucose were lower, and those of glucagon were higher in the food-restricted/deprived groups (INS, both 2.7; DeltaINS, 0.1-0.3 mIU/L; GLN, 2.8-3.3; DeltaGLN, 1.4-2.2 microg/L) than in the control group (means +/- SE; INS, 3.8 +/- 0.8; DeltaINS, 1.4 +/- 0.7 mIU/L; GLN, 2.5 +/- 0.5; DeltaGLN, 0.7 +/- 0.5 microg/L), resulting in similar increases in the insulin:glucagon ratio. Impaired insulin secretion may thus be compensated for by a greater glucagon response and the decreased glucose utilization rates of fasting garden warblers may result from insulin resistance and/or delayed glucose absorption. PMID- 10375471 TI - Vascular angiotensin II receptor and calcium signaling in toadfish. AB - The renin-angiotensin system evolved during the early evolution of vertebrates and regulates blood pressure/blood volume homeostasis in nonmammalian and mammalian vertebrates. Properties of vascular angiotensin (ANG) receptors and signal pathways in primitive animals are, however, not well understood. We aimed to determine whether vascular ANG II receptors in the toadfish, Opsanus tau, an aglomerular teleost, pharmacologically resemble either the ANG subtype 1 receptor (AT1) or the subtype 2 receptor (AT2) by examining (i) the effects of selective ANG receptor antagonists on ANG II-induced vasopressor action and binding and (ii) ANG II's effect on cytosolic Ca2+ signaling. [Asn1, Val5]ANG II (native teleost ANG II) dose-dependently increased the mean arterial pressure of conscious toadfish. ANG II-induced pressor responses (100-500 ng/kg) were inhibited substantially (79-83%) by [Sar1, Ile8]ANG II (5 microg x kg-1 + 5 microg x kg-1 x min-1) and moderately (34-53%) by losartan (AT1 antagonist, 10 mg/kg + 20 mg x kg-1 x h-1) and by PD 123319 (AT2 antagonist, 10 mg/kg + 20 mg x kg-1 x h-1) (36-60%). Likewise, the [Asp1, Val5, His9]ANG I-induced pressor effect was completely eliminated by an ANG I-converting enzyme inhibitor, SQ 14,225. Specific 125I-ANG II binding to vascular smooth muscle (VSM) membrane fractions was displaced completely by [Asn1, Val5]ANG II and [Sar1, Ile8]ANG II. Losartan, but not PD 123319, partly displaced ANG II binding at 10(-10)-10(-6) M. Furthermore, ANG II (10(-7) or 10(-8) M) caused a rapid, transient increase in the cytosolic Ca2+ signal (fluorescence ratio (FR) of 340/380 nm) of isolated VSM tissues measured by fura-2 and a dual wavelength fluorospectrometer, whereas extracellular K+ induced sustained, dose-dependent (P < 0.01) increases in FR. The results indicate that toadfish VSM tissues possess a rather nonselective ANG receptor; partial inhibition of ANG II binding by losartan and stimulation of cytosolic Ca2+ signaling by ANG II suggest that the receptor has some resemblance to AT1 homologous receptors. PMID- 10375472 TI - Concentration-dependent, biphasic effect of prostaglandins on avian corticosteroidogenesis in vitro. AB - Previous work with mammalian and frog adrenocortical tissue and cells indicates that prostaglandins (PGs) can directly stimulate corticosteroidogenesis. However, work with avian adrenal preparations is absent. Therefore, the present studies with isolated chicken (Gallus gallus domesticus) and turkey (Meleagris gallopavo) adrenal steroidogenic cells were conducted to determine whether PGs can directly influence avian corticosteroidogenesis as well. Cells (1 x 10(5) cells/ml) were incubated with a wide range of concentrations of PGs in the presence of indomethacin (1 microg/ml) (to attenuate endogenous PG production) and 1-methyl-3 isobutylxanthine (0.5 mM) [to preserve cyclic AMP (cAMP)] for 2 h. Corticosterone and cAMP production were measured by highly specific radioimmunoassay. PGI2 was without effect. With the exception of PGF2alpha, which had a slight stimulation in chicken but not in turkey cells, the influence of the other PGs on corticosterone production was biphasic. For the stimulatory phase (up to a concentration of 5 x 10(-5) M), there were prostanoid structural and avian species differences in both potency and efficacy of PGs. Overall, PGs were 11 times more potent in turkey cells than in chicken cells. However, the order of potency for stimulation was similar for both chicken and turkey cells: for chicken cells the order was PGE2 > PGE1 > PGA1 > PGB2 > PGB1 > PGF2alpha and for turkey cells it was PGE2 > PGE1 > PGA1 > PGB2 = PGB1. In contrast, PG efficacy for stimulation was greater for chicken cells. In addition, the orders of efficacy were different from the orders of potency. In chicken cells, the order of efficacy was PGE2 = PGA1 > PGE1 > PGB2 > PGB1 > PGF2alpha and for turkey cells it was PGB2 = PGE2 > PGA1 > PGE1 > PGB1. Because of the greater maximal corticosterone response over basal production of chicken cells to PGs, they were used to assess the interaction of PGs with ACTH and to examine more fully the inhibitory phase of PGs. Cells were incubated with PGs in the presence of threshold (2.5 x 10(-11) M), half-maximal (1 x 10(-10) M), and maximal (1 x 10( 7) M) steroidogenic concentrations of ACTH. With the exception of PGF2alpha, the average efficacy of PGs to elevate corticosterone was increased 55% by a threshold steroidogenic concentration of ACTH. However, with higher concentrations of ACTH, this enhancement of efficacy disappeared as did the stimulatory effect of some PGs. The results suggest that the steroidogenic actions of PGs and ACTH converge on the same pool of steroidogenic enzymes leading to corticosterone. At concentrations greater than 5 x 10(-5) M, several PGs (notably PGA1, PGA2, PGB1, and PGB2) inhibited both ACTH-induced and basal corticosterone production. PGA1 and PGA2 were the most potent inhibitors. Corticosterone and cAMP production were closely associated in the biphasic action of PGs, suggesting that the effect of PGs was mediated by the changing levels of intracellular cAMP. Collectively, these data suggest that PGs may be important modulators of corticosteroidogenesis in the avian adrenal gland. PMID- 10375473 TI - Insulin and proglucagon-derived peptides from the horned frog, Ceratophrys ornata (Anura:Leptodactylidae). AB - Insulin and peptides derived from the processing of proglucagon have been isolated from an extract of the pancreas of the South American horned frog, Ceratophrys ornata (Leptodactylidae). Ceratophrys insulin is identical to the insulin previously isolated from the toad, Bufo marinus (Bufonidae). Ceratophrys glucagon was isolated in two molecular forms with 29- and 36-amino acid residues in approximately equal amounts. Glucagon-29 is identical to glucagon from B. marinus and from the bullfrog, Rana catesbeiana (Ranidae) and contains only 1 amino acid substitution (Thr29 --> Ser) compared with glucagon from Xenopus laevis (Pipidae). Glucagon-36 comprises glucagon-29 extended from its C-terminus by Lys-Arg-Ser-Gly-Gly-Met-Ser. This extension is structurally dissimilar to the C-terminal octapeptide of mammalian oxyntomodulin and resembles more closely that found in C-terminally extended glucagons isolated from fish pancreata. Ceratophrys glucagon-like peptide-1 (GLP-1) (His-Ala-Asp-Gly-Thr-Tyr-Gln-Asn-Asp Val10-Gln-Gln-Phe-Leu-Glu- Glu-Lys-Ala-Ala-Lys20-Glu-Phe-Ile-Asp-Trp-Leu-Ile-Lys Gly- Lys30-Pro-Lys-Lys-Gln-Arg-Leu-Ser) contains 3 amino acid substitutions compared with the corresponding peptide from B. marinus, 8 substitutions compared with GLP-1 from R. catesbeiana, and between 4 and 11 substitutions compared with the three GLP-1 peptides identified in X. laevis proglucagon. GLP-2 was not identified in the extract of Ceratophrys pancreas. The data indicate that, despite its importance in the regulation of glucose metabolism, the primary structure of GLP-1 has been very poorly conserved during evolution, even among a single order such as the Anura. PMID- 10375474 TI - The brain-pituitary-gonadal axis of female rainbow trout Oncorhynchus mykiss: effects of photoperiod manipulation1. AB - Two groups of post-spawned female rainbow trout were exposed to two different photoperiods, an ambient photoperiod (56 degrees N) and a combination of long and short photoperiods (a constant 18L:6D from February 1 until May 10, then a constant 6L:18D), which acted to advance maturation and spawning. The stimulatory long-short photoperiod advanced spawning by 3-4 months and correspondingly advanced peaks in serum levels of 17beta-estradiol, testosterone, calcium (an index of vitellogenin), and GTH II. Earlier events in gonadal recrudescence appeared to be less affected by the photoperiod. The initiation of exogenous vitellogenesis coincided with high levels of both pituitary salmon gonadotropin releasing hormone (sGnRH) content and serum follicle-stimulating hormone (FSH, GTH I) levels. High levels of serum FSH were associated with rapid gonadal growth in the fish exposed to the stimulatory long-short photoperiod. In contrast, the fish exposed to the ambient photoperiod showed gonadal steroid production, formation of vitellogenin, and secondary oocyte growth without any detectable increase in serum FSH levels. The possible roles and interactions of sGnRH, gonadotropins, and steroids with respect to normal and artificially stimulated ovarian maturation are discussed. PMID- 10375475 TI - Diaphyseal cross-sectional geometry of the Boxgrove 1 Middle Pleistocene human tibia. AB - Cross-sectional geometric analysis of the early Middle Pleistocene human tibia from Boxgrove, West Sussex, U.K. reveals a mosaic pattern relative to other archaic Homo tibiae. The specimen has relatively low percent cortical area within its cross sections. However, it exhibits the high mediolateral strength characteristic of archaic Homo tibiae. Scaled solely to tibial length it is robust, similar to those of the Neandertals and above those of early modern and pre-Late Pleistocene African and Asian humans. However, given ecogeographically patterned variance in relative tibial length and body laterality, it is most likely that it exhibits a level of robusticity within the range encompassed by Late Pliocene to Late Pleistocene archaic Homo combined with arctic body proportions. Given its association with late interglacial cool temperate climatic indicators, the inferred body proportions of the Boxgrove hominid were probably promoted by their minimal level of cultural buffering, requiring a significant biological conservation of body heat. PMID- 10375476 TI - Middle Palaeolithic burial is not a dead issue: the view from Qafzeh, Saint Cesaire, Kebara, Amud, and Dederiyeh. AB - Inferences of purposeful Middle Palaeolithic (MP) burial are almost universally accepted, despite published arguments that the pre-1960s discoveries are equally well explained by natural processes. In the modern human origins debate (perhaps the most hotly disputed question in palaeoanthropology) inferences of MP burial are crucial in arguments for an early Upper Pleistocene emergence of modern humans. The present paper contributed to that debate by re-examining a number of post-1960s excavations of MP hominid remains. Because these were excavated with meticulous attention to depositional circumstances and stratigraphic context, most palaeoanthropologists consider these inferences of purposeful burial to be based on irrefutable evidence. This paper focuses on the reasoning behind such claims, especially the assumption that articulated sketetal material is prima facie evidence for deliberate burial. First it reviews a range of processes operating in caves and rockshelters that condition the probability of articulated skeletal material preserving without hominid intervention. Processes such as deposition, decomposition, and disturbance are inherently more variable in caves and rockshelters than is usually acknowledged. The first section concludes that purposeful protection is not necessary to account for the preservation of articulated skeletal remains. The second part of the paper examines the published record from Qafzeh, Saint-Cesaire, Kebara, Amud and Dederiyeh, where the majority of the remains claimed to have been buried are fragmented, incomplete, and disarticulated. This re-examination suggests that in all of the post-1960s cases of putative burial, the hominid remains occur in special depositional circumstances, which by themselves are sufficient to account for the preservation in evidence at these sites. This conclusion severely weakens arguments for purposeful burial at the five sites. Moreover, the equivocal nature of the evidence in the more recent cases renders even less secure the similar claims made for discoveries of hominid skeletal remains at La Chapelle-aux-Saints, Le Mousterier, La Ferrassie, Teshik-Tash, La Grotte du Regourdou, Shanidar, and several others. Finally, by highlighting the equivocal nature of the evidence, this paper underscores the ongoing need for palaeoanthropologists to specify as wide a range of taphonomic processes as possible when interpreting the archaeological record. This will aid in producing robust inferences, and will bring about increasingly accurate knowledge of when hominids became human. PMID- 10375477 TI - Aurignacian lithic economy and early modern human mobility: new perspectives from classic sites in the Vezere valley of France. AB - During the past decade the chronology and hominin attributions of the Aurignacian have been revised or called into question. These controversies have coincided with an increased appreciation for the social complexity of Aurignacian culture in the realms of organic technologies and mobiliary and parietal manifestations of symbolic behavior. Lithic raw material procurement and reduction intensity evidence from Aurignacian occupations at the Vezere Valley sites of Abri Pataud, Le Facteur, and La Ferrassie may reflect complex group mobility strategies. The lithic components under consideration were always dominated by cherts available within a few kilometers radius. Assemblages associated with the early Aurignacian have elevated proportions of cherts from distant sources. Lithic retouch data indicate that some early Aurignacian assemblages reflect greater extent and/or intensity of marginal retouch compared with the later Aurignacian. Lithic reduction data, however, reveal evidence of greater core reduction intensity during the later Aurignacian. Flexible strategies of residential mobility, possibly in response to changes in the subsistence environment, may account for some of the variability between early and later Aurignacian assemblages. Similar shifts in raw material procurement were evidently associated with the Middle Paleolithic in southwestern France. However, Aurignacian populations may have acquired most lithic materials by movement directly to sources, while certain non utilitarian materials were probably obtained via some form of indirect social exchange. This suggested coexistence of direct and indirect procurement mechanisms serves to distinguish Aurignacian assemblages from earlier Middle Paleolithic deposits and emphasizes that socially-directed intensification was one of the fundamental elements of the suite of cultural changes referred to as the Middle-Upper Paleolithic transition. PMID- 10375478 TI - The first Paranthropus from the Malawi Rift. PMID- 10375479 TI - Modern human origins as seen from the peripheries. PMID- 10375480 TI - Intraoperative duplex imaging of carotid endarterectomy. PMID- 10375481 TI - The validity of ultrasonographic scanning as screening method for abdominal aortic aneurysm. AB - OBJECTIVE: the sensitivity and specificity of screening for abdominal aortic aneurysms (AAAs) with ultrasonographic scanning (US) is unknown. The aim of the study was to validate US as screening test for AAAs. METHODS AND MATERIAL: 4176 (76.3%) of 5470 men aged 65-73 attended hospital-based US screening for an AAA at their local hospital. Two observers and one scanner were used. The maximal anterior-posterior (AP) of the dilated aorta, or 2 cm above the bifurcation, and at the crossing of left renal vein was recorded. In 50 cases, blinded measurements were carried out by two observers. An AAA was defined as an AP diameter greater than 29 mm. RESULTS: the standard deviation (s.d.) of the interobserver variability of the distal AP diameter was 0.84. The mean distal AP diameter was 17. 9 mm (s.d. 2.92). Combining these data, the estimated diagnostic sensitivity was 98.9%, the estimated diagnostic specificity was 99. 9%. The interobserver s.d. of the proximal AP diameter was 1.76. The mean proximal AP diameter was 18.4 mm (s.d. 2.45). Combining these data, the estimated diagnostic sensitivity was 87.4%, the estimated diagnostic specificity was 99.9%. CONCLUSION: US seems to be a valid screening method for AAA. Screening for proximal infrarenal aorta aneurysm remains acceptable because the majority of aortic diameters in this segment are so much smaller than the diameters that define an AAA. PMID- 10375482 TI - Computer analysis of ultrasonic plaque echolucency in identifying high risk carotid bifurcation lesions. AB - OBJECTIVES: to confirm that plaque echogenicity evaluated by computer analysis, as suggested by preliminary studies, can identify plaques associated with a high incidence of strokes. MATERIALS AND METHODS: a series of 96 patients with carotid stenosis in the range of 50-99% were studied retrospectively (41 with TIAs and 55 asymptomatic). Carotid plaque echogenicity was evaluated using a computerised measurement of the median grey scale value (GSM). All patients had a CT brain scan to determine the presence of infarction in the carotid territory. RESULTS: the incidence of ipsilateral brain CT infarctions was 16% in the asymptomatic and 32% in the symptomatic plaques (p =0.076). It was 20% for <70% stenosis and 25% for >70% stenosis (p =0.52). It was 9% for plaques which had a GSM >50 and 40% in those with GSM <50 (p <0.001) with a relative risk of 4.6 (95% CI 1.8 to 11.6). CONCLUSIONS: the results confirm that computer analysis of plaque echogenicity is better than the degree of stenosis in identifying plaques associated with an increased incidence of CT brain-scan infarction and consequently useful for identifying individuals at high risk of stroke. What is required is a form of image standardisation in order to apply this method to natural history studies with stroke as the endpoint. PMID- 10375483 TI - Angiographic runoff score as a predictor of outcome following femorocrural bypass surgery. AB - OBJECTIVE: to evaluate the efficacy of the revised ad hoc scoring system in predicting the outcome of femorocrural bypass surgery. DESIGN: retrospective study. MATERIALS AND METHODS: seventy-seven infrainguinal bypass procedures to the crural arteries were performed in 69 patients with critical leg ischaemia. Preoperative angiographic findings were graded according to the revised ad hoc scoring system and other preoperative angiographic measures. RESULTS: the revised ad hoc scores were valuable in predicting the outcome of these grafts. The status of the outflow artery throughout its length had a great impact on the long-term outcome in terms of secondary patency, leg salvage, patients alive with legs, and survival rates. In situ autogenous saphenous grafts achieved the best immediate and long-term results. CONCLUSIONS: the revised ad hoc angiographic scoring method is useful in predicting the outcome of patients undergoing femorocrural arterial reconstruction. Patients with an outflow artery completely open throughout its length had excellent long-term results. PMID- 10375484 TI - Influence of the collateral function of the circle of Willis on hemispherical perfusion during carotid occlusion as assessed by transcranial colour-coded duplex ultrasonography. AB - OBJECTIVES: to investigate the collateral potential of the circle of Willis with transcranial colour-coded duplex ultrasonography and common carotid artery (CCA) compression. MATERIALS AND METHODS: in 46 atherosclerotic patients without cerebrovascular disease, the functional patency of the collaterals of the circle of Willis, the anterior and posterior communicating arteries, was assessed. The Peak Systolic Velocity (PSV) decrease in the middle cerebral artery (MCA) during CCA compression between complete and incomplete circles was compared. RESULTS: in 10 (22%) patients a complete and in 36 (78%) patients an incomplete circle of Willis was found, mainly due to non-functioning posterior communicating arteries. In hemispheres with collateral supply through both the anterior and the posterior communicating artery, the median PSV decrease in the MCA during CCA compression was 43%. When the posterior, anterior or both communicating arteries (1 hemisphere) were missing the PSV decrease was 58% (p =0.003), 70% (p =0.001) and 75%, respectively. CONCLUSIONS: collateral flow from the basilar to the carotid territory is often hampered by non-functioning posterior communicating arteries. A non-functioning anterior communicating artery is rare. A complete collateral circulation provides better perfusion of the MCA during carotid occlusion as compared with collateral supply through only the anterior or the posterior communicating artery in the case of an incomplete circle of Willis. PMID- 10375485 TI - Controlling transplant vasculopathy in cryopreserved vein grafts with polyethylene glycol and glutathione during transport. AB - BACKGROUND: the biological characteristics of cryopreserved allografts are poorly understood, although many factors are known to influence their outcome. This study examines the development of transplant vasculopathy in both fresh and cryopreserved vein allografts and specifically assesses the efficacy of a transport solution containing 10% polyethylene glycol and 10 microM glutathione (PEG/GSH). METHODS: jugular veins were harvested from control donor rabbits and transplanted as interposition carotid bypass grafts in 30 New Zealand White (NZW) rabbits. Ten received the fresh jugular veins (fresh). Ten animals received jugular veins which had been harvested, transported in a physiological solution, cryopreserved and stored in a standard fashion (cryopreserved). Ten animals received jugular veins which had been harvested, transported in the same solution with the addition of PEG/GSH, cryopreserved and stored in a standard fashion (PEG/GSH). Cryopreserved jugular veins were stored for 6 weeks before transplantation. All animals were sacrificed 28 days postoperatively. Vein grafts were perfusion-fixed and wall dimensions were determined by planimetry. RESULTS: all transplanted grafts were patent at harvest. The control cryopreserved vein grafts showed a 54% increase in mean intimal thickness (63+/-10 micron vs. 41+/-3 micron p<0.05) but no change in mean medial thickness (125+/-9 micron vs. 119+/ 13 micron; p = N.S. ) compared to the fresh allograft. Transport of the grafts in PEG/GSH solution resulted in the abolition of the increase in intimal thickness (41+/-4 micron; p <0.01) associated with cryopreservation without a change in medial thickness (140+/-15 micron; p = N.S.) compared to the cryopreserved allograft. CONCLUSION: cryopreserved vein grafts develop significant intimal hyperplasia compared to freshly transplanted grafts. The use of PEG/GSH in the transport solution significantly reduces this transplant graft intimal hyperplasia to that which develops in fresh grafts and may lead to improvements in the clinical use of cryopreserved veins. PMID- 10375486 TI - Are variations in the use of carotid endarterectomy explained by population Need? A study of health service utilisation in two English health regions. AB - OBJECTIVES: to describe variation in utilisation of carotid endarterectomy (CEA) within two English health regions and explore relationships between use, need and proximity to services. DESIGN: consecutive case series of operations. Comparison at a population level with district stroke mortality, hospital admissions and material deprivation. MAIN OUTCOME MEASURES: standardised utilisation rates for CEA and measures of inter-district variability. Spearman's rank correlation coefficients for associations between variables. RESULTS: variation in utilisation rates was considerable (14-fold difference across district populations). More individuals had bilateral surgery in the Yorkshire region than in the Northern (11.7% vs. 5.5%, p=0.002). There was no association between utilisation rates for CEA and district stroke mortality (r=-0.06, 95% CI -0.41 to 0.30) or admission rates for stroke (r=0.17, 95% CI -0.2 to 0.49). There was a strong relationship between residence in districts where services were located and higher utilisation. Rates of CEA were lowest in the regions' most affluent wards. CONCLUSION: use of CEA varies widely, depending on area of residence. Variation is not a consequence of differences in need, but reflects clinical practice and supply of services. There is evidence to suggest unmet need for CEA. PMID- 10375487 TI - The effect of 'non-critical' (<50%) stenosis on vein graft longitudinal resistance and impedance. AB - OBJECTIVE: vein graft stenoses <50% cause minimal flow impairment, velocity elevation, or symptomatology and are therefore usually assumed to be "non critical". The purpose of this study was to assess the effect of <50% vein graft stenosis on vein graft longitudinal impedance, as elevated impedance has been found to correlate with clinical graft failure. METHODS: eight segments of non reversed cryopreserved vein (mean length 23+/-1 cm; mean outer diameter 4.7+/-0.2 mm) were saline-perfused in vitro utilising a variable pulsatile perfusion pump, Windkessel, and clamp resistor simulating the haemodynamic conditions of arterial bypass. Proximal (Pprox) and distal (Pdist) pressure were continuously measured by fluid-filled catheter transduction, and flow (Q) by ultrasonic transit-time flowmetry. Waveforms were digitally recorded at 200 Hz at pulse rates ranging from 60-180 b.p.m. with mean flow (Q) of 154 ml/min and mean proximal pressure (Pprox) of 100 mmHg (max/min 120/90). Graded mid-graft stenoses of <50% were created using an inflatable vascular occluder and measured by the corresponding changes in mean pressure gradient (DeltaP=Pprox-Pdist) and Q (%stenosis=1 {DeltaPbaselineQstenosis/Delta PstenosisQbaseline}1/4). Vein graft longitudinal resistance (RL) was calculated as DeltaP/Q. After Fourier transformation, vein graft longitudinal impedance (ZL) was calculated as DeltaP/Q at each harmonic, with ZL determined by integration over 0-4 Hz. Results are reported as mean+/ S.E.M. RESULTS: the desired levels of pressure and flow were established in all vein segments. Graded inflation of the occluder resulted in vein graft stenosis of 23+/-3% and 39+/-3%. This was accompanied by a mild reduction in Q (12% and 30%) and considerable increases in both RL (180% and 710%) and ZL (140% and 430%). CONCLUSIONS: "non-critical" vein graft stenosis (<50%) causes minimal change in mean flow, but substantial elevations in longitudinal resistance and impedance. The contribution of "non-critical" stenosis to vein graft failure may be under-appreciated. PMID- 10375488 TI - Studies in calf venous pump function utilizing a two-valve experimental model. AB - OBJECTIVES: to explore the hydrodynamic mechanisms involved in the regulation of ambulatory venous pressure. DESIGN: an experimental model of calf venous pump was constructed with collapsible tubes and valves. MATERIAL: the model consisted of a conduit and a pump with an intervening competent valve. Another valve that could allow reflux into the pump was mounted above the pump. METHODS: conduit pressure and recovery times were monitored under conditions of different degrees of ejection fraction and reflux into the pump. Model variables included using poorly compliant tubes for the pump, the conduit and for both the pump and conduit. RESULTS: the latex tube exhibited a non-linear volume-pressure relationship and a bi-modal regimen of compliance. This bestowed pressure-buffering properties. Ambulatory venous hypertension resulted when reflux beyond buffering capacity occurred. Substituting less compliant PTFE for latex at the pump had a relatively minor effect on post-ejection pressure and recovery times. Using PTFE at the conduit had a profound but divergent effect on both of these parameters. Conduit capacitance reduction had a similar effect. CONCLUSION: conduit elastance plays a significant role in the regulation of ambulatory venous pressure in this experimental model. The hydrodynamic principles illustrated by the model may enhance our understanding of the human calf venous pump. PMID- 10375489 TI - Vitamin C reduces ischaemia-reperfusion-induced acute lung injury. AB - OBJECTIVES: to evaluate vitamin C supplementation in the prevention of ischaemia reperfusion (I-R) induced acute lung injury. DESIGN: Sprague-Dawley rats (n =6/group) were randomised into Control, I-R and I-R pretreated with vitamin C (3.3 g over 5 days). Ischaemia-reperfusion injury was induced by 30 minutes infrarenal aortic cross-clamping and 120 minutes reperfusion. METHODS: pulmonary microvascular injury was measured by broncho-alveolar lavage protein concentration, pulmonary neutrophil infiltration by tissue myeloperoxidase activity and bronchoalveolar lavage neutrophil counts. In a second experiment (n =5/group) neutrophil respiratory burst activity was measured in Control and vitamin C groups. RESULTS: ischaemia-reperfusion resulted in a significant increase in both microvascular leakage and pulmonary neutrophil infiltration as measured by bronchoalveolar lavage protein concentration and pulmonary myeloperoxidase activity respectively. Pretreatment with vitamin C significantly attenuated both microvascular leakage and neutrophil infiltration. Neutrophil respiratory burst activity was significantly reduced in the vitamin C group (13.02 m.c.f.+/-0.3) compared with Control (19.04 m.c.f.+/-1. 9),p <0.02. CONCLUSION: these data suggest that oral vitamin C therapy protects against ischaemia-reperfusion-induced acute lung injury, possibly by attenuating neutrophil respiratory burst activity. PMID- 10375490 TI - Vascular and endovascular surgical activity in Denmark, Finland, Norway, Sweden and Northern Ireland. PMID- 10375491 TI - Persistent sciatic vein - unusual cause of reflux from the popliteal fossa and sural nerve damage. PMID- 10375495 TI - Volume contents PMID- 10375492 TI - Loneliness prior to CABG. PMID- 10375496 TI - The genetics of cell migration in Drosophila melanogaster and Caenorhabditis elegans development. AB - Cell migrations are found throughout the animal kingdom and are among the most dramatic and complex of cellular behaviors. Historically, the mechanics of cell migration have been studied primarily in vitro, where cells can be readily viewed and manipulated. However, genetic approaches in relatively simple model organisms are yielding additional insights into the molecular mechanisms underlying cell movements and their regulation during development. This review will focus on these simple model systems where we understand some of the signaling and receptor molecules that stimulate and guide cell movements. The chemotactic guidance factor encoded by the Caenorhabditis elegans unc-6 locus, whose mammalian homolog is Netrin, is perhaps the best known of the cell migration guidance factors. In addition, receptor tyrosine kinases (RTKs), and FGF receptors in particular, have emerged as key mediators of cell migration in vivo, confirming the importance of molecules that were initially identified and studied in cell culture. Somewhat surprisingly, screens for mutations that affect primordial germ cell migration in Drosophila have revealed that enzymes involved in lipid metabolism play a role in guiding cell migration in vivo, possibly by producing and/or degrading lipid chemoattractants or chemorepellents. Cell adhesion molecules, such as integrins, have been extensively characterized with respect to their contribution to cell migration in vitro and genetic evidence now supports a role for these receptors in certain instances in vivo as well. The role for non-muscle myosin in cell motility was controversial, but has now been demonstrated genetically, at least in some cell types. Currently the best characterized link between membrane receptor signaling and regulation of the actin cytoskeleton is that provided by the Rho family of small GTPases. Members of this family are clearly essential for the migrations of some cells; however, key questions remain concerning how chemoattractant and chemorepellent signals are integrated within the cell and transduced to the cytoskeleton to produce directed cell migration. New types of genetic screens promise to fill in some of these gaps in the near future. PMID- 10375497 TI - Role of PDGF-B and PDGFR-beta in recruitment of vascular smooth muscle cells and pericytes during embryonic blood vessel formation in the mouse. AB - Development of a vascular system involves the assembly of two principal cell types - endothelial cells and vascular smooth muscle cells/pericytes (vSMC/PC) - into many different types of blood vessels. Most, if not all, vessels begin as endothelial tubes that subsequently acquire a vSMC/PC coating. We have previously shown that PDGF-B is critically involved in the recruitment of pericytes to brain capillaries and to the kidney glomerular capillary tuft. Here, we used desmin and alpha-smooth muscle actin (ASMA) as markers to analyze vSMC/PC development in PDGF-B-/- and PDGFR-beta-/- embryos. Both mutants showed a site-specific reduction of desmin-positive pericytes and ASMA-positive vSMC. We found that endothelial expression of PDGF-B was restricted to immature capillary endothelial cells and to the endothelium of growing arteries. BrdU labeling showed that PDGFR beta-positive vSMC/PC progenitors normally proliferate at sites of endothelial PDGF-B expression. In PDGF-B-/- embryos, limb arterial vSMC showed a reduced BrdU labeling index. This suggests a role of PDGF-B in vSMC/PC cell proliferation during vascular growth. Two modes of vSMC recruitment to newly formed vessels have previously been suggested: (1) de novo formation of vSMC by induction of undifferentiated perivascular mesenchymal cells, and (2) co-migration of vSMC from a preexisting pool of vSMC. Our data support both modes of vSMC/PC development and lead to a model in which PDGFR-beta-positive vSMC/PC progenitors initially form around certain vessels by PDGF-B-independent induction. Subsequent angiogenic sprouting and vessel enlargement involves PDGF-B-dependent vSMC/PC progenitor co-migration and proliferation, and/or PDGF-B-independent new induction of vSMC/PC, depending on tissue context. PMID- 10375498 TI - Characterization of the transvection mediating region of the abdominal-B locus in Drosophila. AB - Genetic studies have identified an unusual transvection process in the Abdominal B (Abd-B) locus of Drosophila. In some cases distal infraabdominal (iab) regulatory domains continue to activate the Abd-B promoter even when translocated onto different chromosomes. Transvection depends on an approx. 10 kb genomic DNA sequence, termed the transvection mediating region (tmr), located immediately downstream of the Abd-B transcription unit. Here we report a detailed analysis of this region. Different DNA fragments from the tmr were inserted into a variety of P-transformation vectors. Analyses of reporter gene expression in transgenic embryos and adults identify at least three cis-regulatory elements, including two enhancers (IAB7 and IAB8) and a new insulator DNA (Frontabdominal-8, Fab-8). Evidence is also presented for a Polycomb Response Element (PRE) linked to the IAB8 enhancer, and an internal promoter in the iab-8 domain, which transcribes the iab-7 and iab-8 cis-regulatory DNA, including the Fab-8 insulator. We discuss the significance of these findings with regard to Abd-B transvection and long range enhancer-promoter interactions in mammalian globin loci. PMID- 10375499 TI - Induction of the mesendoderm in the zebrafish germ ring by yolk cell-derived TGF beta family signals and discrimination of mesoderm and endoderm by FGF. AB - The endoderm forms the gut and associated organs, and develops from a layer of cells which emerges during gastrula stages in the vertebrate embryo. In comparison to mesoderm and ectoderm, little is known about the signals which induce the endoderm. The origin of the endoderm is intimately linked with that of mesoderm, both by their position in the embryo, and by the molecules that can induce them. We characterised a gene, zebrafish gata5, which is expressed in the endoderm from blastula stages and show that its transcription is induced by signals originating from the yolk cell. These signals also induce the mesoderm expressed transcription factor no tail (ntl), whose initial expression coincides with gata5 in the cells closest to the blastoderm margin, then spreads to encompass the germ ring. We have characterised the induction of these genes and show that ectopic expression of activin induces gata5 and ntl in a pattern which mimics the endogenous expression, while expression of a dominant negative activin receptor abolishes ntl and gata5 expression. Injection of RNA encoding a constitutively active activin receptor leads to ectopic expression of gata5 and ntl. gata5 is activated cell-autonomously, whereas ntl is induced in cells distant from those which have received the RNA, showing that although expression of both genes is induced by a TGF-beta signal, expression of ntl then spreads by a relay mechanism. Expression of a fibroblast growth factor (eFGF) or a dominant negatively acting FGF receptor shows that ntl but not gata5 is regulated by FGF signalling, implying that this may be the relay signal leading to the spread of ntl expression. In embryos lacking both squint and cyclops, members of the nodal group of TGF-beta related molecules, gata5 expression in the blastoderm is abolished, making these factors primary candidates for the endogenous TGF-beta signal inducing gata5. PMID- 10375500 TI - Extragenic suppressors of the arabidopsis zwi-3 mutation identify new genes that function in trichome branch formation and pollen tube growth. AB - The plant cytoskeleton plays a pivotal role in determining the direction of cell wall expansion, and ultimately the cell's final shape. However, the mechanisms by which localized expansion events are initiated remain obscure. Mutational analysis of the trichome (plant hair) morphogenic pathway in Arabidopsis has identified at least eight genes that determine trichome branch number. One of these genes, ZWICHEL (ZWI), encodes a novel member of the kinesin superfamily of motor proteins. Mutations in the ZWI gene cause a reduction in the number of trichome branches. To identify additional genes involved in trichome branch initiation, we screened for extragenic suppressors of the zwi-3 mutation and isolated three suppressors that rescued the branch number defect of zwi-3. These suppressors define three genes, named suz, for suppressor of zwichel-3. All of the suppressors were shown to be allele specific. One of the suppressors, suz2, also rescued the trichome branch number defect of another branch mutant, furca1 2. Plants homozygous for suz2 have more than the wild-type number of trichome branches. This suggests that SUZ2 is a negative regulator of trichome branching and may interact with ZWI and FURCA1. The suz1 and suz3 mutants display no obvious phenotype in the absence of the zwi-3 mutation. The suz1 zwi-3 double mutants also exhibited a male-sterile phenotype due to a defect in pollen tube germination and growth, whereas both the suz1 and the zwi-3 single mutants are fertile. The synthetic male sterility of the suz1 zwi-3 double mutants suggests a role for SUZ1 and ZWI in pollen germination and pollen tube growth. DNA sequence analysis of the zwi-3 mutation indicated that only the tail domain of the zwi-3 protein would be expressed. Thus, the suz mutations show allele-specific suppression of a kinesin mutant that lacks the motor domain. PMID- 10375501 TI - Sonic hedgehog regulates the growth and patterning of the cerebellum. AB - The molecular bases of brain development and CNS malignancies remain poorly understood. Here we show that Sonic hedgehog (Shh) signaling controls the development of the cerebellum at multiple levels. SHH is produced by Purkinje neurons, it is required for the proliferation of granule neuron precursors and it induces the differentiation of Bergmann glia. Blocking SHH function in vivo results in deficient granule neuron and Bergmann glia differentiation as well as in abnormal Purkinje neuron development. Thus, our findings provide a molecular model for the growth and patterning of the cerebellum by SHH through the coordination of the development of cortical cerebellar cell types. In addition, they provide a cellular context for medulloblastomas, childhood cancers of the cerebellum. PMID- 10375502 TI - Phases of cytoplasmic and cortical reorganizations of the ascidian zygote between fertilization and first division. AB - Many eggs undergo reorganizations that localize determinants specifying the developmental axes and the differentiation of various cell types. In ascidians, fertilization triggers spectacular reorganizations that result in the formation and localization of distinct cytoplasmic domains that are inherited by early blastomeres that develop autonomously. By applying various imaging techniques to the transparent eggs of Phallusia mammillata, we now define 9 events and phases in the reorganization of the surface, cortex and the cytoplasm between fertilization and first cleavage. We show that two of the domains that preexist in the egg (the ER-rich cortical domain and the mitochondria-rich subcortical myoplasm) are localized successively by a microfilament-driven cortical contraction, a microtubule-driven migration and rotation of the sperm aster with respect to the cortex, and finally, a novel microfilament-dependant relaxation of the vegetal cortex. The phases of reorganization we have observed can best be explained in terms of cell cycle-regulated phases of coupling, uncoupling and recoupling of the motions of cortical and subcortical layers (ER-rich cortical domain and mitochondria-rich domain) with respect to the surface of the zygote. At the end of the meiotic cell cycle we can distinguish up to 5 cortical and cytoplasmic domains (including two novel ones; the vegetal body and a yolk-rich domain) layered against the vegetal cortex. We have also analyzed how the myoplasm is partitioned into distinct blastomeres at the 32-cell stage and the effects on development of the ablation of precisely located small fragments. On the basis of our observations and of the ablation/ transplantation experiments done in the zygotes of Phallusia and several other ascidians, we suggest that the determinants for unequal cleavage, gastrulation and for the differentiation of muscle and endoderm cells may reside in 4 distinct cortical and cytoplasmic domains localized in the egg between fertilization and cleavage. PMID- 10375503 TI - The role of tolloid/mini fin in dorsoventral pattern formation of the zebrafish embryo. AB - A highly conserved TGF-&bgr; signaling pathway is involved in the establishment of the dorsoventral axis of the vertebrate embryo. Specifically, Bone Morphogenetic Proteins (Bmps) pattern ventral tissues of the embryo while inhibitors of Bmps, such as Chordin, Noggin and Follistatin, are implicated in dorsal mesodermal and neural development. We investigated the role of Tolloid, a metalloprotease that can cleave Chordin and increase Bmp activity, in patterning the dorsoventral axis of the zebrafish embryo. Injection of tolloid mRNA into six dorsalized mutants rescued only one of these mutants, mini fin. Through chromosomal mapping, linkage and cDNA sequence analysis of several mini fin alleles, we demonstrate that mini fin encodes the tolloid gene. Characterization of the mini fin mutant phenotype reveals that Mini fin/Tolloid activity is required for patterning ventral tissues of the tail: the ventral fin, and the ventroposterior somites and vasculature. Gene expression studies show that mfn mutants exhibit reduced expression of ventrally restricted markers at the end of gastrulation, suggesting that the loss of ventral tail tissues is caused by a dorsalization occurring at the end of gastrulation. Based on the mini fin mutant phenotype and the expression of tolloid, we propose that Mini fin/Tolloid modifes the Bmp activity gradient at the end of gastrulation, when the ventralmost marginal cells of the embryo are in close proximity to the dorsal Chordin expressing cells. At this time, unimpeded Chordin may diffuse to the most ventral marginal regions and inhibit high Bmp activity levels. In the presence of Mini fin/Tolloid, however, Chordin activity would be negatively modulated through proteolytic cleavage, thereby increasing Bmp signaling activity. This extracellular mechanism is amplified by an autoregulatory loop for bmp gene expression. PMID- 10375504 TI - Precise developmental regulation of Ets family transcription factors during specification and commitment to the T cell lineage. AB - Ets family transcription factors control the expression of a large number of genes in hematopoietic cells. Here we show strikingly precise differential expression of a subset of these genes marking critical, early stages of mouse lymphocyte cell-type specification. Initially, the Ets family member factor Erg was identified during an arrayed cDNA library screen for genes encoding transcription factors expressed specifically during T cell lineage commitment. Multiparameter fluorescence-activated cell sorting for over a dozen cell surface markers was used to isolate 18 distinct primary-cell populations representing discrete T cell and B cell developmental stages, pluripotent lymphoid precursors, immature NK-like cells and myeloid hematopoietic cells. These populations were monitored for mRNA expression of the Erg, Ets-1, Ets-2, Fli-1, Tel, Elf-1, GABPalpha, PU.1 and Spi-B genes. The earliest stages in T cell differentiation show particularly dynamic Ets family gene regulation, with sharp transitions in expression correlating with specification and commitment events. Ets, Spi-B and PU.1 are expressed in these stages but not by later T-lineage cells. Erg is induced during T-lineage specification and then silenced permanently, after commitment, at the beta-selection checkpoint. Spi-B is transiently upregulated during commitment and then silenced at the same stage as Erg. T-lineage commitment itself is marked by repression of PU.1, a factor that regulates B-cell and myeloid genes. These results show that the set of Ets factors mobilized during T-lineage specification and commitment is different from the set that maintains T cell gene expression during thymocyte repertoire selection and in all classes of mature T cells. PMID- 10375505 TI - Antagonism of EGFR and notch signalling in the reiterative recruitment of Drosophila adult chordotonal sense organ precursors. AB - The selection of Drosophila melanogaster sense organ precursors (SOPs) for sensory bristles is a progressive process: each neural equivalence group is transiently defined by the expression of proneural genes (proneural cluster), and neural fate is refined to single cells by Notch-Delta lateral inhibitory signalling between the cells. Unlike sensory bristles, SOPs of chordotonal (stretch receptor) sense organs are tightly clustered. Here we show that for one large adult chordotonal SOP array, clustering results from the progressive accumulation of a large number of SOPs from a persistent proneural cluster. This is achieved by a novel interplay of inductive epidermal growth factor-receptor (EGFR) and competitive Notch signals. EGFR acts in opposition to Notch signalling in two ways: it promotes continuous SOP recruitment despite lateral inhibition, and it attenuates the effect of lateral inhibition on the proneural cluster equivalence group, thus maintaining the persistent proneural cluster. SOP recruitment is reiterative because the inductive signal comes from previously recruited SOPs. PMID- 10375506 TI - XCtBP is a XTcf-3 co-repressor with roles throughout Xenopus development. AB - XTcf-3 is an HMG box transcription factor that mediates Xenopus dorsal-ventral axis formation. As a Wnt pathway effector, XTcf-3 interacts with beta-catenin and activates the expression of the dorsal organizing gene siamois, while in the absence of beta-catenin, XTcf-3 functions as a transcriptional repressor. We show that XTcf-3 contains amino- and carboxy-terminal repressor domains and have identified a Xenopus member of the C-terminal Binding Protein family of transcriptional co-repressors (XCtBP) as the C-terminal co-repressor. We show that two XCtBP binding sites near the XTcf-3 carboxy-terminus are required for the interaction of XTcf-3 and XCtBP and for the transcriptional repression mediated by the XTcf-3 carboxy-terminal domain. By fusing the GAL4 activation domain to XCtBP we have generated an antimorphic protein, XCtBP/G4A, that activates siamois transcription through an interaction with endogenous XTcf-3. Ectopic expression of XCtBP/G4A demonstrates that XCtBP functions in the regulation of head and notochord development. Our data support a role for XCtBP as a co-repressor throughout Xenopus development and indicate that XCtBP/G4A will be a useful tool in determining how XCtBP functions in various developmental processes. PMID- 10375507 TI - UNC-84 localizes to the nuclear envelope and is required for nuclear migration and anchoring during C. elegans development. AB - Nuclear migrations are essential for metazoan development. Two nuclear migrations that occur during C. elegans development require the function of the unc-84 gene. unc-84 mutants are also defective in the anchoring of nuclei within the hypodermal syncytium and in the migrations of the two distal tip cells of the gonad. Complementation analyses of 17 unc-84 alleles defined two genetically separable functions. Both functions are required for nuclear and distal tip cell migrations, but only one is required for nuclear anchorage. The DNA lesions associated with these 17 mutations indicate that the two genetically defined functions correspond to two distinct regions of the UNC-84 protein. The UNC-84 protein has a predicted transmembrane domain and a C-terminal region with similarity to the S. pombe spindle pole body protein Sad1 and to two predicted mammalian proteins. Analysis of a green fluorescent protein reporter indicated that UNC-84 is widely expressed and localized to the nuclear envelope. We propose that UNC-84 functions to facilitate a nuclear-centrosomal interaction required for nuclear migration and anchorage. PMID- 10375508 TI - A short region of its homeodomain is necessary for engrailed nuclear export and secretion. AB - Engrailed homeoprotein, a transcription factor involved in midbrain/hindbrain patterning, primarily localizes to the cell nucleus. However, significant amounts of the protein are also found in the cell cytoplasm or associated with membrane microdomains enriched in cholesterol and glycosphingoglycolipids (Joliot, A., Trembleau, A., Raposo, G., Calvet, S., Volovitch, M. and Prochiantz, A. (1997) Development 124, 1865-1875). This non-nuclear localization, observed in vitro and in vivo, led us to investigate the possibility that Engrailed be transferred between nuclear and non-nuclear compartments. Monkey COS-7 cells expressing chick Engrailed-2 (cEN2) were fused with 3T3 mouse fibroblasts and the passage of cEN2 from COS-7 to 3T3 nuclei was followed in the interspecies heterokaryons. We find that, 10 minutes following cell fusion, cEN2 is detected in the 3T3 nuclei of 80% of the heterokaryons demonstrating rapid cEN2 nuclear export. Export from donor nuclei can be saturated and is strongly reduced after deletion of a 11 amino acid long (&Dgr;)1 sequence present within a slightly larger domain that extends between helices 2 and 3 of the homeodomain and shows strong similarities with leucine-rich nuclear export signals (NES). This putative NES, when fused with a nuclear reporter protein, allows its nuclear export, demonstrating that it is not only necessary but also sufficient for nuclear export and can therefore be considered as a true nuclear export sequence. In an earlier report (Joliot, A., Maizel, A., Rosenberg, D., Trembleau, A., Dupas, S., Volovitch, M. and Prochiantz, A. (1998) Current Biology 8, 856-863), we demonstrated that the (&Dgr;)1 sequence is necessary for the access of cEN2 to the lumen of a membrane compartment and for its intercellular transfer. The present study thus strongly suggests that the regulation of Engrailed nuclear export could play a role not only in Engrailed transcriptional activity but also in its ability to gain access to a secretory compartment. PMID- 10375509 TI - Fgf8 and Gbx2 induction concomitant with Otx2 repression is correlated with midbrain-hindbrain fate of caudal prosencephalon. AB - Chick/quail transplantation experiments were performed to analyse possible factors involved in the regionalisation of the midbrain-hindbrain domain. The caudal prosomeres, expressing Otx2, were transplanted at stage HH10 into rostrocaudal levels of the midbrain-hindbrain domain, either straddling the intra metencephalic constriction (type 1 grafts), or at rostral and medial levels of pro-rhombomere A1 (type 2 and 3 grafts, respectively); thus, in all situations, one border of the graft was in contact with the host Gbx2- and Fgf8-expressing domains. The area containing the graft, recognised by QCPN immunohistochemistry, was first analysed 48 hours after transplantation for Otx2, Gbx2, En2 and Fgf8. Although in all three situations, a large part of the graft maintained Otx2 expression, another part became Otx2 negative and was induced to express Gbx2 and Fgf8. These inductive events occurred exclusively at the interface between the Otx2-positive transplanted domain and the ipsilateral host Gbx2-positive rhombomere 1, creating a new Otx2-Gbx2 boundary within the grafted territory. In type 1 and 2 grafts, the induced Fgf8 domain is in continuity with the host Fgf8 isthmic domain, whereas for type 3 grafts, these two domains are separate. High levels of En2 expression were also induced in the area expressing Gbx2 and Fgf8, and Wnt1 and Pax2 expressions, analysed in type 3 grafts, were induced at the intragraft Otx2-Gbx2 new boundary. Moreover, at later embryonic stages, the graft developed meso-isthmo-cerebellar structures. Thus, gene expressions induced in the grafted prosencephalon not only mimicked the pattern observed in the normal midbrain-hindbrain domain, but is followed by midbrain-hindbrain cytodifferentiation, indicating that not only Fgf8 but also confrontation of Otx2 and Gbx2 may play an essential role during midbrian-hindbrain regionalisation. PMID- 10375510 TI - Gli proteins encode context-dependent positive and negative functions: implications for development and disease. AB - Several lines of evidence implicate zinc finger proteins of the Gli family in the final steps of Hedgehog signaling in normal development and disease. C-terminally truncated mutant GLI3 proteins are also associated with human syndromes, but it is not clear whether these C-terminally truncated Gli proteins fulfil the same function as full-length ones. Here, structure-function analyses of Gli proteins have been performed using floor plate and neuronal induction assays in frog embryos, as well as induction of alkaline phosphatase (AP) in SHH-responsive mouse C3H10T1/2 (10T1/2) cells. These assays show that C-terminal sequences are required for positive inducing activity and cytoplasmic localization, whereas N terminal sequences determine dominant negative function and nuclear localization. Analyses of nuclear targeted Gli1 and Gli2 proteins suggest that both activator and dominant negative proteins are modified forms. In embryos and COS cells, tagged Gli cDNAs yield C-terminally deleted forms similar to that of Ci. These results thus provide a molecular basis for the human Polydactyly type A and Pallister-Hall Syndrome phenotypes, derived from the deregulated production of C terminally truncated GLI3 proteins. Analyses of full-length Gli function in 10T1/2 cells suggest that nuclear localization of activating forms is a regulated event and show that only Gli1 mimics SHH in inducing AP activity. Moreover, full length Gli3 and all C-terminally truncated forms act antagonistically whereas Gli2 is inactive in this assay. In 10T1/2 cells, protein kinase A (PKA), a known inhibitor of Hh signaling, promotes Gli3 repressor formation and inhibits Gli1 function. Together, these findings suggest a context-dependent functional divergence of Gli protein function, in which a cell represses Gli3 and activates Gli1/2 prevents the formation of repressor Gli forms to respond to Shh. Interpretation of Hh signals by Gli proteins therefore appears to involve a fine balance of divergent functions within each and among different Gli proteins, the misregulation of which has profound biological consequences. PMID- 10375511 TI - A role for fasciclin II in the guidance of neuronal migration. AB - The insect cell adhesion receptor fasciclin II is expressed by specific subsets of neural and non-neural cells during embryogenesis and has been shown to control growth cone motility and axonal fasciculation. Here we demonstrate a role for fasciclin II in the guidance of migratory neurons. In the developing enteric nervous system of the moth Manduca sexta, an identified set of neurons (the EP cells) undergoes a stereotyped sequence of migration along the visceral muscle bands of the midgut prior to their differentiation. Probes specific for Manduca fasciclin II show that while the EP cells express fasciclin II throughout embryogenesis, their muscle band pathways express fasciclin II only during the migratory period. Manipulations of fasciclin II in embryonic culture using blocking antibodies, recombinant fasciclin II fragments, and enzymatic removal of glycosyl phosphatidylinositol-linked fasciclin II produced concentration dependent reductions in the extent of EP cell migration. These results support a novel role for fasciclin II, indicating that this homophilic adhesion molecule is required for the promotion or guidance of neuronal migration. PMID- 10375512 TI - Xenopus nodal-related signaling is essential for mesendodermal patterning during early embryogenesis. AB - Previously, we showed that Xenopus nodal-related factors (Xnrs) can act as mesoderm inducers, and that activin induces Xnr transcription, suggesting that Xnrs relay or maintain induction processes initiated by activin-like molecules. We used a dominant negative cleavage mutant Xnr2 (cmXnr2) to carry out loss-of function experiments to explore the requirement for Xnr signaling in early amphibian embryogenesis, and the relationship between activin and Xnrs. cmXnr2 blocked mesoderm induction caused by Xnr, but not activin, RNA. In contrast, cmXnr2 did suppress mesoderm and endoderm induction by activin protein, while Xnr transcript induction was unaffected by cmXnr2, consistent with an interference with the function of Xnr peptides that were induced by activin protein treatment. The severe hyperdorsalization and gastrulation defects caused by Xnr2 in whole embryos were rescued by cmXnr2, establishing a specific antagonistic relationship between the normal and cleavage mutant proteins. Expression of cmXnr2 resulted in delayed dorsal lip formation and a range of anterior truncations that were associated with delayed and suppressed expression of markers for dorsoanterior endoderm, in which the recently recognized head organizer activity resides. Reciprocally, Xnr2 induced dorsoanterior endodermal markers, such as cerberus, Xhex-1 and Frzb, in animal cap ectoderm. The migratory behavior of head mesendoderm explanted from cmXnr2 RNA-injected embryos was drastically reduced. These results indicate that Xnrs play crucial roles in initiating gastrulation, probably by acting downstream of an activin-like signaling pathway that leads to dorsal mesendodermal specification, including setting up the head organizer. PMID- 10375513 TI - The Drosophila homeobox genes zfh-1 and even-skipped are required for cardiac specific differentiation of a numb-dependent lineage decision. AB - A series of inductive signals are necessary to subdivide the mesoderm in order to allow the formation of the progenitor cells of the heart. Mesoderm-endogenous transcription factors, such as those encoded by twist and tinman, seem to cooperate with these signals to confer correct context and competence for a cardiac cell fate. Additional factors are likely to be required for the appropriate specification of individual cell types within the forming heart. Similar to tinman, the zinc finger- and homeobox-containing gene, zfh-1, is expressed in the early mesoderm and later in the forming heart, suggesting a possible role in heart development. Here, we show that zfh-1 is specifically required for formation of the even-skipped (eve)-expressing subset of pericardial cells (EPCs), without affecting the formation of their siblings, the founders of a dorsal body wall muscle (DA1). In addition to zfh-1, mesodermal eve itself appears to be needed for correct EPC differentiation, possibly as a direct target of zfh-1. Epistasis experiments show that zfh-1 specifies EPC development independently of numb, the lineage gene that controls DA1 founder versus EPC cell fate. We discuss the combinatorial control mechanisms that specify the EPC cell fate in a spatially precise pattern within the embryo. PMID- 10375514 TI - Regulation of midline development by antagonism of lefty and nodal signaling. AB - The embryonic midline is crucial for the development of embryonic pattern including bilateral symmetry and left-right asymmetry. In zebrafish, lefty1 (lft1) and lefty2 (lft2) have distinct midline expression domains along the anteroposterior axis that overlap with the expression patterns of the nodal related genes cyclops and squint. Altered expression patterns of lft1 and lft2 in zebrafish mutants that affect midline development suggests different upstream pathways regulate each expression domain. Ectopic expression analysis demonstrates that a balance of lefty and cyclops signaling is required for normal mesendoderm patterning and goosecoid, no tail and pitx2 expression. In late somite-stage embryos, lft1 and lft2 are expressed asymmetrically in the left diencephalon and left lateral plate respectively, suggesting an additional role in laterality development. A model is proposed by which the vertebrate midline, and thus bilateral symmetry, is established and maintained by antagonistic interactions among co-expressed members of the lefty and nodal subfamilies of TGF beta signaling molecules. PMID- 10375515 TI - Starvation promotes Dictyostelium development by relieving PufA inhibition of PKA translation through the YakA kinase pathway. AB - When nutrients are depleted, Dictyostelium cells undergo cell cycle arrest and initiate a developmental program that ensures survival. The YakA protein kinase governs this transition by regulating the cell cycle, repressing growth-phase genes and inducing developmental genes. YakA mutants have a shortened cell cycle and do not initiate development. A suppressor of yakA that reverses most of the developmental defects of yakA- cells, but none of their growth defects was identified. The inactivated gene, pufA, encodes a member of the Puf protein family of translational regulators. Upon starvation, pufA- cells develop precociously and overexpress developmentally important proteins, including the catalytic subunit of cAMP-dependent protein kinase, PKA-C. Gel mobility-shift assays using a 200-base segment of PKA-C's mRNA as a probe reveals a complex with wild-type cell extracts, but not with pufA- cell extracts, suggesting the presence of a potential PufA recognition element in the PKA-C mRNA. PKA-C protein levels are low at the times of development when this complex is detectable, whereas when the complex is undetectable PKA-C levels are high. There is also an inverse relationship between PufA and PKA-C protein levels at all times of development in every mutant tested. Furthermore, expression of the putative PufA recognition elements in wild-type cells causes precocious aggregation and PKA-C overexpression, phenocopying a pufA mutation. Finally, YakA function is required for the decline of PufA protein and mRNA levels in the first 4 hours of development. We propose that PufA is a translational regulator that directly controls PKA-C synthesis and that YakA regulates the initiation of development by inhibiting the expression of PufA. Our work also suggests that Puf protein translational regulation evolved prior to the radiation of metazoan species. PMID- 10375516 TI - Transcription factors: the right combination for the DNA lock. AB - Recently determined structures of complexes between homeodomain proteins, their cofactors and DNA have provided new insights into the way pairs of transcription factors can collaborate to select the appropriate target DNA-binding sites during development. PMID- 10375517 TI - Taste processing: whetting our appetites. AB - Two G-protein-coupled receptors have been identified that are present in the apical membranes of rat and mouse taste cells and differentially distributed across the tongue and palate. They are strong candidates for being taste receptors and their discovery has provided new tools for research into gustatory processing. PMID- 10375518 TI - Cell cycle: driving the centrosome cycle. AB - Cyclin-dependent kinases (Cdks) control major transitions as cells pass through the cell cycle. It has recently been shown that centrosome duplication in vertebrates requires Cdk2 activity and can be driven solely by Cdk2-cyclin E complexes. PMID- 10375519 TI - Insulin secretion: feed-forward control of insulin biosynthesis? AB - It has long been accepted wisdom that insulin secreted from islet beta cells has either no effect, or an inhibitory feedback effect, on insulin synthesis and secretion. Recent work suggests, instead, that secreted insulin acts directly on beta cells, via its own receptor, to enhance insulin production in an autocrine feed-forward loop. PMID- 10375520 TI - Axon guidance: starting and stopping with slit. AB - As developing axons navigate, they exhibit various behaviours: extending and branching, pausing, changing direction, retracting. Now, the Slit protein has been discovered to have striking positive and negative effects on axon growth and guidance. PMID- 10375521 TI - Nuclear transport: randy couples. AB - The recently solved structures of the Ran GTPase with a Ran-binding domain and with karyopherin-beta2 have revealed unusually tight embraces that provide important insights into the mechanism of nuclear transport and the many ways in which common protein folds are adapted to perform very different functions. PMID- 10375522 TI - Genetics: a touch of elegance with RNAi. AB - RNA-mediated gene interference (RNAi), a rapid, convenient tool for inhibiting gene function in Caenorhabditis elegans, has recently been shown to work in other organisms. PMID- 10375523 TI - Dna repair: Rad52 - the means to an end. AB - In eukaryotic organisms, double-strand breaks in chromosomal DNA are repaired either by non-homologous end-joining, or by homologous recombination. How do cells choose which pathway to use? PMID- 10375524 TI - HIV: a new role for Nef in the spread of HIV. AB - The HIV Nef protein downregulates the cell-surface expression of the HIV receptor glycoprotein CD4, but the significance of this event has remained obscure. Recent data suggest that Nef reduces cell-surface CD4 to promote the efficient spread of the virus. PMID- 10375525 TI - Inhibition of HIV-1 progeny virion release by cell-surface CD4 is relieved by expression of the viral Nef protein. AB - BACKGROUND: The human immunodeficiency virus type 1 (HIV-1) Nef protein is required for efficient virus replication in vivo and displays a number of distinct and apparently unrelated biological activities in vitro. Of these, one of the most readily demonstrated is the efficient internalization and degradation of cell-surface CD4, the receptor for the HIV-1 envelope protein. The biological purpose of this internalization has, however, remained unclear. RESULTS: Using human 293T cells expressing high levels of cell-surface CD4 or CD8, we demonstrate that CD4, but not CD8, can dramatically reduce the release of infectious virions bearing the HIV-1 envelope protein and induce a concomitant increase in the accumulation of cell-associated HIV-1 structural proteins. In contrast, CD4 had no effect on the release of HIV-1 bearing a heterologous envelope protein unable to bind CD4. Nef expression totally reversed CD4-mediated inhibition but only if the CD4 used remained susceptible to Nef-induced internalization. CONCLUSIONS: These results support the hypothesis that cell surface CD4 can interact with the envelope protein present on budding HIV-1 virions to inhibit their release. The internalization and degradation of cell surface CD4 induced by the viral Nef protein can fully reverse this inhibition and is, therefore, likely to facilitate the spread of virus in vivo. PMID- 10375526 TI - The COE transcription factor Collier is a mediator of short-range Hedgehog induced patterning of the Drosophila wing. AB - BACKGROUND: The secreted Hedgehog (Hh) proteins have been implicated as mediators of positional information in vertebrates and invertebrates. A gradient of Hh activity contributes to antero-posterior (A/P) patterning of the fly wing. In addition to inducing localised expression of Decapentaplegic (Dpp), which in turn relays patterning cues at long range, Hh directly patterns the central region of the wing. RESULTS: We show that short-range, dose-dependent Hh activity is mediated by activation of the transcription factor Collier (Col). In the absence of col activity, longitudinal veins 3 and 4 (L3 and L4) are apposed and the central intervein is missing. Hh expression induces col expression in a narrow stripe of cells along the A/P boundary through a dual-input mechanism: inhibition of proteolysis of Cubitus-interruptus (Ci) and activation of the Fused (Fu) kinase. Col, in cooperation with Ci, controls the formation of the central intervein by activating the expression of blistered (bs), which encodes the Drosophila serum response factor (D-SRF), the activity of which is required for the adoption and maintenance of the intervein cell fate. Furthermore, col is allelic to knot, a gene involved in the formation of the central part of the wing. This finding completes our understanding of the sectorial organisation of the Drosophila wing. CONCLUSIONS: Col, the Drosophila member of the COE family (Col/Olf-1/EBF) of non-basic, helix-loop-helix (HLH)-containing transcription factors, is a mediator of the short-range organising activity of Hh in the Drosophila wing. Our results support the idea that Hh controls target gene expression in a concentration-dependent manner and highlight the importance of the Fu kinase in this differential regulation. The high degree of evolutionary conservation of the COE proteins and the diversity of developmental processes controlled by Hh signalling raises the possibility that the specific genetic interactions depicted here may also operate in vertebrates. PMID- 10375527 TI - Interplay between Rac and Rho in the control of substrate contact dynamics. AB - BACKGROUND: Substrate anchorage and cell locomotion entail the initiation and development of different classes of contact sites, which are associated with the different compartments of the actin cytoskeleton. The Rho-family GTPases are implicated in the signalling pathways that dictate contact initiation, maturation and turnover, but their individual roles in these processes remain to be defined. RESULTS: We monitored the dynamics of peripheral, Rac-induced focal complexes in living cells in response to perturbations of Rac and Rho activity and myosin contractility. We show that focal complexes formed in response to Rac differentiated into focal contacts upon upregulation of Rho. Focal complexes were dissociated by inhibitors of myosin-II-dependent contractility but not by an inhibitor of Rho-kinase. The downregulation of Rac promoted the enlargement of focal contacts, whereas a block in the Rho pathway not only caused a dissolution of focal contacts but also stimulated membrane ruffling and formation of new focal complexes, which were associated with the advance of the cell front. CONCLUSIONS: Rac functions to signal the creation of new substrate contacts at the cell front, which are associated with the induction of ruffling lamellipodia, whereas Rho serves in the maturation of existing contacts, with both contact types requiring contractility for their formation. The transition from a focal complex to a focal contact is associated with a switch to Rho-kinase dependence. Rac and Rho also influence the development of focal contacts and focal complexes, respectively, through mutually antagonistic pathways. PMID- 10375528 TI - Cell-surface expression of CD4 reduces HIV-1 infectivity by blocking Env incorporation in a Nef- and Vpu-inhibitable manner. AB - BACKGROUND: Human immunodeficiency virus-1 (HIV-1) infection decreases the cell surface expression of its cellular receptor, CD4, through the combined actions of Nef, Env and Vpu. Such functional convergence strongly suggests that CD4 downregulation is critical for optimal viral replication, yet the significance of this phenomenon has so far remained a puzzle. RESULTS: We show that high levels of CD4 on the surface of HIV-infected cells induce a dramatic reduction in the infectivity of released virions by the sequestering of the viral envelope by CD4. CD4 is able to accumulate in viral particles while at the same time blocking incorporation of Env into the virion. Nef and Vpu, through their ability to downregulate CD4, counteract this effect. CONCLUSIONS: The CD4-mediated 'envelope interference' described here probably explains the plurality of mechanisms developed by HIV to downregulate the cell-surface expression of its receptor. PMID- 10375529 TI - A new member of the endonuclease III family of DNA repair enzymes that removes methylated purines from DNA. AB - DNA is constantly exposed to endogenous andexogenous alkylating agents that can modify its bases,resulting in mutagenesis in the absence of DNA repair [1,2]. Alkylation damage is removed by the action of DNA glycosylases, which initiate the base excision repair pathway and protect the sequence information of the genome [3-5]. We have identified a new class of methylpurine DNA glycosylase, designated MpgII, that is a member of the endonuclease III family of DNA repair enzymes. We expressed and purified MpgII from Thermotoga maritima and found that the enzyme releases both 7-methylguanine and 3-methyladenine from DNA. We cloned the MpgII genes from T. maritima and from Aquifex aeolicus and found that both genes could restore methylmethanesulfonate (MMS) resistance to Escherichia coli alkA tagA double mutants, which are deficient in the repair of alkylated bases. Analogous genes are found in other Bacteria and Archaea and appear to be the only genes coding for methylpurine DNA glycosylase activity in these organisms. MpgII is the fifth member of the endonuclease III family of DNA repair enzymes, suggesting that the endonuclease III protein scaffold has been modified during evolution to recognize and repair a variety of DNA damage. PMID- 10375530 TI - Mad2 binding by phosphorylated kinetochores links error detection and checkpoint action in mitosis. AB - The spindle checkpoint must detect the presence of unattached or improperly attached kinetochores and must then inhibit progression through the cell cycle until the offending condition is resolved. Detection probably involves attachment sensitive kinetochore phosphorylation (reviewed in [1,2]). A key player in the checkpoint's response is the Mad2 protein, which prevents activation of the anaphase-promoting complex (APC) by the Cdc20 protein [3-8]. Microinjection of Mad2 antibodies results in premature anaphase onset [9,10], and excess Mad2 protein causes arrest in mitosis [5,11]. We have previously shown that Mad2 localizes to unattached kinetochores in vertebrate cells, and that this localization ceases as kinetochores accumulate microtubules [10,12,13]. But how is Mad2 binding limited to unattached kinetochores? Here, we used lysed PtK1 cells to study kinetochore phosphorylation and Mad2 binding. We found that Mad2 binds to phosphorylated kinetochores, but not to unphosphorylated ones. Our data suggest that it is kinetochore protein phosphorylation that promotes Mad2 binding to unattached kinetochores. Thus, we have identified a probable molecular link between attachment-sensitive kinetochore phosphorylation and the inhibition of anaphase. The complete pathway for error control in mitosis can now be outlined. PMID- 10375531 TI - Inhibition of Rho at different stages of thymocyte development gives different perspectives on Rho function. AB - Development of thymocytes can be staged according to the levels of expression of the cell-surface markers CD4, CD8, CD44, CD25 and CD2. Thymocyte development is regulated by a complex signalling network [1], one component of which is the GTPase Rho. The bacterial enzyme C3 transferase from Clostridium botulinum selectively ADP-ribosylates Rho in its effector-binding domain and thereby abolishes its biological function [2,3]. To explore the function of Rho in thymocyte development, we previously used the proximal promoter of the gene encoding the Src-family kinase p56lck to make transgenic mice that selectively express C3 transferase in the thymus [4,6]. In these mice, which lack Rho function from the earliest thymocyte stages, thymocyte numbers are reduced by approximately 50- to 100-fold. Here, we describe transgenic mice that express C3 transferase under the control of the locus control region (LCR) of the CD2 gene; this regulatory element drives expression at a later stage of thymocyte development than the lck proximal promoter [7]. In these mice, thymocyte numbers were also reduced by 50- to 100-fold, but unlike the lck-C3 mice, in which the reduction predominantly results from defects in cell survival of CD25(+) thymocyte progenitors, the CD2-C3 transgenic mice had a pre-T-cell differentiation block at the CD25(+) stage after rearrangement of the T-cell receptor (TCR) beta chains. Analysis of CD2-C3 mice demonstrated that Rho acts as an intracellular switch for TCR beta selection, the critical thymic differentiation checkpoint. These results show that Rho-mediated survival signals for CD25(+) pre-T cells are generated by the extracellular signals that act on earlier thymocyte precursors and also that temporal cell-type-specific elimination of Rho can reveal different functions of this GTPase in vivo. PMID- 10375533 TI - O sole mio! PMID- 10375532 TI - p27(Kip1) ubiquitination and degradation is regulated by the SCF(Skp2) complex through phosphorylated Thr187 in p27. AB - Many tumorigenic processes affect cell-cycle progression by their effects on the levels of the cyclin-dependent kinase inhibitor p27(Kip1) [1,2]. The phosphorylation- and ubiquitination-dependent proteolysis of p27 is implicated in control of the G1-S transition in the cell cycle [3-6]. To determine the factors that control p27 stability, we established a cell-free extract assay that recapitulates the degradation of p27. Phosphorylation of p27 at Thr187 was essential for its degradation. Degradation was also dependent on SCF(Skp2), a protein complex implicated in targeting phosphorylated proteins for ubiquitination [7-10]. Immunodepletion of components of the complex - Cul-1, Skp1, or Skp2 - from the extract abolished p27 degradation, while addition of purified SCF(Skp2) to Skp2- depleted extract restored the capacity to degrade p27. A specific association was observed between Skp2 and a p27 carboxy-terminal peptide containing phosphorylated Thr187, but not between Skp2 and the non phosphorylated peptide. Skp2-dependent associations between Skp1 or Cul-1 and the p27 phosphopeptide were also detected. Isolated SCF(Skp2) contained an E3 ubiquitin ligase activity towards p27. Our data thus suggest that SCF(Skp2) specifically targets p27 for degradation during cell-cycle progression. PMID- 10375534 TI - Salvador moncada: NO holding back PMID- 10375535 TI - Pain. PMID- 10375537 TI - Aquatic jewels PMID- 10375536 TI - Focal adhesions. PMID- 10375538 TI - Synthesis and application of functionally diverse 2,6,9-trisubstituted purine libraries as CDK inhibitors. AB - BACKGROUND: Purines constitute a structural class of protein ligands involved in mediating an astonishing array of metabolic processes and signal pathways in all living organisms. Synthesis of purine derivatives targeting specific purine binding proteins in vivo could lead to versatile lead compounds for use as biological probes or drug candidates. RESULTS: We synthesized several libraries of 2,6, 9-trisubstituted purines using both solution- and solid-phase chemistry, and screened the compounds for inhibition of cyclin-dependent kinase (CDK) activity and human leukemic cell growth. Lead compounds were optimized by iterative synthesis based on structure-activity relationships (SARs), as well as analysis of several CDK-inhibitor cocrystal structures, to afford several interesting compounds including one of the most potent CDK inhibitors known to date. Unexpectedly, some compounds with similar CDK inhibitory activity arrested cellular proliferation at distinctly different phases of the cell cycle and another inhibitor directly induced apoptosis, bypassing cell-cycle arrest. Some of these compounds selectively inhibited growth of cells derived from specific tumors. CONCLUSIONS: 2,6,9-Trisubstituted purines have various and potent biological activities, despite high concentrations of competing endogenous purine ligands in living cells. Purine libraries constitute a versatile source of small molecules that affect distinct biochemical pathways mediating different cellular functions. PMID- 10375539 TI - The binding site for an inhibitor of squalene:hopene cyclase determined using photoaffinity labeling and molecular modeling. AB - BACKGROUND: The squalene:hopene cyclases (SHCs) are bacterial enzymes that convert squalene into hopanoids, a function analogous to the action of oxidosqualene cyclases (OSCs) in eukaryotic steroid and triterpenoid biosynthesis. We have identified the binding site for a selective, potent, photoactivatable inhibitor of an SHC. RESULTS: SHC from Alicyclobacillus acidocaldarius was specifically labeled by [3H]Ro48-8071, a benzophenone containing hypocholesteremic drug. Edman degradation of a peptide fragment of covalently modified SHC confirmed that Ala44 was specifically modified. Molecular modeling, using X-ray-derived protein coordinates and a single point constraint for the inhibitor, suggested several geometries by which Ro48-8071 could occupy the active site. CONCLUSIONS: A covalent complex of a potent inhibitor with a squalene cyclase has been characterized. The amino acid modification and molecular modeling suggest that Ro48-8071 binds at the junction between the central cavity and substrate entry channel, therefore inhibiting access of the substrate to the active site. PMID- 10375540 TI - A unique urinary constituent, 6-hydroxy-6-methyl-3-heptanone, is a pheromone that accelerates puberty in female mice. AB - BACKGROUND: Olfactorily mediated puberty acceleration in female mice (measured by an increase in uterine weight) has been observed since the 1960s without the active chemosignal being structurally identified. There are many controversies in the literature as to whether this male-originated pheromone is a volatile substance. We investigated the chemical nature of the urinary fractions that are responsible for the characteristic uterine weight increases. RESULTS: The active pheromone was identified as 5,5-dimethyl-2-ethyltetrahydrofuran-2-ol and/or its open-chain tautomer (6-hydroxy-6-methyl-3-heptanone). A series of cyclic vinyl ethers were isolated from chromatographically active fractions of the urine. Because these compounds did not accelerate puberty, we postulated that these ethers were degradation products of a lactol (5,5-dimethyl-2-ethyltetrahydrofuran 2-ol). The lactol was then detected directly in the mouse urine extract using a silylation agent. Synthetic 6-hydroxy-6-methyl-3-heptanone had strong biological activity, whereas its close structural analogs did not. CONCLUSIONS: The male house mouse excretes into its urine a large quantity of a volatile substance that has a unique lactol/hydroxyketone structure. This substance is capable of binding to the less volatile urinary constituents, such as proteins or peptides, and is active in puberty-acceleration bioassays. The controversies regarding the volatility of the puberty-accelerating pheromones can now be explained by considering a complex of volatile lactol/hydroxyketone and urinary proteins. PMID- 10375541 TI - Binding of a dimeric derivative of vancomycin to L-Lys-D-Ala-D-lactate in solution and at a surface. AB - BACKGROUND: The emergence of bacteria that are resistant to vancomycin (V), a glycopeptide antibiotic, results from the replacement of the carboxy-terminal D Ala-D-Ala of bacterial cell wall precursors by D-Ala-D-lactate. Recently, it has been demonstrated that covalent dimeric variants of V are active against vancomycin-resistant enterococci (VRE). To study the contribution of divalency to the activities of these variants, we modeled the interactions of V and a dimeric V with L-Lys-D-Ala-D-lactate, an analog of the cell-wall precursors of the vancomycin-resistant bacteria. RESULTS: A dimeric derivative of V (V-Rd-V) was found to be much more effective than V in inhibiting the growth of VRE. The interactions of V and V-Rd-V with a monomeric lactate ligand - diacetyl-L-Lys-D Ala-D-lactate (Ac2KDADLac) - and a dimeric derivative of L-Lys-D-Ala-D-lactate (Lac-R'd-Lac) in solution have been examined using isothermal titration calorimetry and UV spectroscopy titrations; the results reveal that V-Rd-V binds Lac-R'd-Lac approximately 40 times more tightly than V binds Ac2KDADLac. Binding of V and of V-Rd-V to Nalpha-Ac-L-Lys-D-Ala-D-lactate presented on the surface of mixed self-assembled monolayers (SAMs) of alkanethiolates on gold indicates that the apparent off-rate for dissociation of V-Rd-V from the surface is much slower than that of V from the same surface. CONCLUSIONS: The results are compatible with the hypothesis that divalency is responsible for tight binding, which correlates with small values of minimum inhibitory concentrations of V and V-Rd V. PMID- 10375542 TI - Assembly line enzymology by multimodular nonribosomal peptide synthetases: the thioesterase domain of E. coli EntF catalyzes both elongation and cyclolactonization. AB - BACKGROUND: EntF is a 142 kDa four domain (condensation-adenylation-peptidyl carrier protein-thioesterase) nonribosomal peptide synthetase (NRPS) enzyme that assembles the Escherichia coli N-acyl-serine trilactone siderophore enterobactin from serine, dihydroxybenzoate (DHB) and ATP with three other enzymes (EntB, EntD and EntE). To assess how EntF forms three ester linkages and cyclotrimerizes the covalent acyl enzyme DHB-Ser-S-PCP (peptidyl carrier protein) intermediate, we mutated residues of the proposed catalytic Ser-His-Asp triad of the thioesterase (TE) domain. RESULTS: The Ser1138-->Cys mutant (kcat decreased 1000-fold compared with wild-type EntF) releases both enterobactin (75%) and linear (DHB-Ser)2 dimer (25%) as products. The His 1271-->Ala mutant (kcat decreased 10,000-fold compared with wild-type EntF) releases only enterobactin, but accumulates both DHB-Ser-O TE and (DHB-Ser)2-O-TE acyl enzyme intermediates. Electrospray ionization and Fourier transform mass spectrometry of proteolytic digests were used to analyze the intermediates. CONCLUSIONS: These results establish that the TE domain of EntF is both a cyclotrimerizing lactone synthetase and an elongation catalyst for ester-bond formation between covalently tethered DHB-Ser moieties, a new function for chain-termination TE domains found at the carboxyl termini of multimodular NRPSs and polyketide synthases. PMID- 10375543 TI - Cellular delivery of peptide nucleic acids and inhibition of human telomerase. AB - BACKGROUND: Human telomerase has an essential RNA component and is an ideal target for developing rules correlating oligonucleotide chemistry with disruption of biological function. Similarly, peptide nucleic acids (PNAs), DNA analogs that bind complementary sequences with high affinity, are outstanding candidates for inducing phenotypic changes through hybridization. RESULTS: We identify PNAs directed to nontemplate regions of the telomerase RNA that can overcome RNA secondary structure and inhibit telomerase by intercepting the RNA component prior to holoenzyme assembly. Relative potencies of inhibition delineate putative structural domains. We describe a novel protocol for introducing PNAs into eukaryotic cells and report efficient inhibition of cellular telomerase by PNAs. CONCLUSIONS: PNAs directed to nontemplate regions are a new class of telomerase inhibitor and may contribute to the development of novel antiproliferative agents. The dependence of inhibition by nontemplate-directed PNAs on target sequence suggests that PNAs have great potential for mapping nucleic acid structure and predictably regulating biological processes. Our simple method for introducing PNAs into cells will not only be useful for probing the complex biology surrounding telomere length maintenance but can be broadly applied for controlling gene expression and functional genomics. PMID- 10375544 TI - Towards a molecular understanding of arthritis. AB - Several different agents including free radicals, oxidizing compounds and proteases are believed to play a role in the onset of arthritis. The evidence and underlying chemistry presently available for each destructive agent are presented. PMID- 10375545 TI - Histidine kinases and two-component signal transduction systems. AB - The phosphorylation of histidine is the first step in many signal transduction cascades in bacteria, yeast and higher plants. The transfer of a very reactive phosphoryl group from phosphorylated histidine kinase to an acceptor is an essential step in many cellular signaling responses. PMID- 10375546 TI - Novel mutant green fluorescent protein protease substrates reveal the activation of specific caspases during apoptosis. AB - BACKGROUND: The caspase-mediated proteolysis of many cellular proteins is a critical event during programmed cell death or apoptosis. It is important to determine which caspases are activated in mammalian cells, and where and when activation occurs, upon receipt of specific death stimuli. Such information would be useful in the design of strategies to regulate the activation of caspases during apoptosis. RESULTS: We developed two novel fluorescent substrates that were specifically cleaved by caspase-1 or caspase-3. For in vitro studies, four amino-acid recognition sequences, YVAD for caspase-1 and DEVD for caspase-3, were introduced between blue fluorescent protein (BFP) and green fluorescent protein (GFP), expressed in bacteria and purified. For in vivo studies, YVAD and DEVD were introduced between cyan fluorescent protein and yellow fluorescent protein, and expression was monitored in live mammalian cells. The proximity between fluorophores was determined using fluorescence resonance energy transfer. Purified substrates were cleaved following exposure to purified caspase-1 and caspase-3. In Cos-7 cells, caspase-1 and caspase-3 substrates were cleaved upon induction of apoptosis with staurosporine, a protein-kinase inhibitor, whereas caspase-3 but not caspase-1 substrate was cleaved upon treatment of cells with the DNA-damaging agent mitomycin c. CONCLUSIONS: These substrates allow the spatial activation of specific members of the caspase family to be deciphered during the initiation and execution phase of programmed cell death, and allow activation of specific caspases to be monitored both in vivo and in vitro. This technology is also likely to be useful for high-throughput screening of reagents that modulate caspase activity. PMID- 10375547 TI - The regulation of antigen-receptor signaling by protein tyrosine phosphatases: a hole in the story. AB - Antigen-receptor signaling requires Src-family kinases to initiate tyrosine phosphorylation. CD45 dephosphorylates the inhibitory site in Src-family kinases before antigen-receptor engagement and thus serves to 'prime' the kinases. It has been unclear why CD45 does not also dephosphorylate 'activated' kinases or motifs within the cytoplasmic domains of antigen-receptors and thus prevent signal transduction. Recent reports raise the possibility that CD45 is excluded from engaged antigen-receptors by mechanisms that may include the formation of lipid microdomains. PMID- 10375548 TI - Cytotoxic T lymphocyte activation by cross-priming. AB - Cross-priming represents the induction of cytotoxic T lymphocyte responses to exogenous, usually cell associated, antigens that are captured and re-presented on bone-marrow-derived antigen-presenting cells. For effective cross-priming, cytotoxic T lymphocytes require help from CD4(+) T cells, which mediate this help indirectly via modification of the antigen-presenting cell. Recent advances made in research into the cellular and molecular interactions required for cross priming have greatly improved our understanding of the antigenic requirements for effective priming and have revealed a role for CD40 and its ligand in the provision of T cell help. PMID- 10375549 TI - T cell co-stimulatory molecules other than CD28. AB - CD28 is the primary co-stimulatory receptor for inducing high-level IL-2 production and survival of naive CD4(+) T cells. While no other cell surface receptor can be considered fully redundant with CD28, recent developments suggest that additional co-stimulatory pathways have preferential effects at different stages of T cell activation, on different subsets of T cells or contribute to the development of different effector functions. PMID- 10375550 TI - Co-receptors on B lymphocytes. AB - Co-receptors have been shown to regulate the antigen-receptor signaling threshold for B cell responses by modulating the activation of signaling molecules that are essential for transmitting a signal through the antigen-receptor. Co-receptors appear to modulate the signaling threshold for B cell tolerance distinctly from that for B cell activation. PMID- 10375551 TI - T cell antigen-receptor signal transduction. AB - During the past year, major progress has been made in understanding proximal TCR signal-transduction events. Cbl has been identified as a negative regulator of kinases from the ZAP-70/Syk family. Studies on LAT, SLP-76, Itk and Vav have revealed their role in the activation of Ras and phospholipase-Cgamma1-Ca2+ signalling pathways. TCR-induced cytoskeletal changes involve signalling through SLP-76-Vav-Nck to activate effectors of the Rho-family of GTPases. Finally, glycolipid-enriched microdomains play a crucial role in T cell activation. PMID- 10375552 TI - Natural killer cell activation and inhibition by receptors for MHC class I. AB - Several recent advances have been made in our understanding of the mechanisms which human natural killer cells recognize MHC class I molecules. Three are of special relevance: the identification of a novel molecule (DAP12) with a key role in the activation pathways; the observation that certain immunoglobulin-like receptors for HLA class I molecules are also utilized by other leucocyte lineages; and the definition of MHC class Ib proteins (i.e. HLA-E and Qa-1b) as specific ligands for the phylogenetically conserved CD94-NKG2 lectin-like receptors. PMID- 10375553 TI - Apoptosis signaling in lymphocytes. AB - Lymphocyte apoptosis is essential for proper function of the immune system. Among other functions, it is responsible for the homeostasis of immune cells and plays a key role in the elimination of autoreactive lymphocytes. Recently, progress has been made in the identification of genes that regulate apoptosis. Studies characterizing the basic apoptosis signaling machinery have begun to reveal the molecular control of processes that modulate lymphocyte apoptosis. PMID- 10375554 TI - Signal transduction from the B cell antigen-receptor. AB - Ligation of the B cell antigen-receptor triggers an intricate maze of intercalated biochemical events that ultimately affect B cell biological responses. Recent advances have helped to connect many loose ends by identifying key adaptor proteins, such as BLNK/SLP-65, defining crucial roles for phosphatidylinositol-3-kinase and mapping pathways controlling the mitogen activated protein kinases (ERK, JNK and p38). PMID- 10375555 TI - Developmental regulation of dendritic cell function. AB - 1998 saw key advances in our understanding of the molecular mechanisms whereby immature dendritic cells recognise foreign pathogens in tissues and are induced to migrate to secondary lymphoid organs. In particular, there have been some key insights into how dendritic cells subsequently direct the evolution of immune responses by differential expression of co-stimulatory molecules. PMID- 10375556 TI - Chemokine receptors in lymphoid organ homeostasis. AB - Leukocytes respond to complex patterns of chemoattractant cytokine (chemokine) gradients that guide them to their destinations in secondary lymphoid organs. This directed movement of multiple cell types requires the choreographed expression of specific G-protein-coupled chemokine receptors and both positive and negative regulation of the signal transduction pathways emanating from them. PMID- 10375557 TI - CTLA-4 and T cell activation. AB - The past year has seen significant advances in our understanding of the role of cytotoxic T lymphocyte antigen 4 (CTLA-4) in regulating T cell activation and tolerance. Recent studies indicate that CTLA-4 not only counterbalances CD28 signals but also can inhibit T cell responses independently of CD28. Recent work has also revealed a role for CTLA-4 in regulating Th1/Th2 differentiation. Manipulation of CTLA-4 in animal models of autoimmunity has shown that CTLA-4 regulates both the initiation and the progression of autoimmune diseases. PMID- 10375558 TI - IL-12 and IFN-gamma in host defense against mycobacteria and salmonella in mice and men. AB - The development of gene-knockout mice and the identification of gene-deficient humans have improved our understanding of the role of IL-12 and IFN-gamma in host defense. Comparison of experimental and natural infections has shown that animals and humans genetically deficient in immunity mediated by IL-12 or IFN-gamma are highly susceptible to mycobacteria and salmonella. Impaired secretion of, or response to, IFN-gamma is the common pathogenic mechanism that accounts for impaired granuloma formation and uncontrolled growth of bacteria within macrophages. The axis formed between IL-12 and IFN-gamma is essential for protective immunity against mycobacteria and salmonella in mice and men. PMID- 10375559 TI - CD1-mediated immune responses to glycolipids. AB - Major advances have recently been accomplished in understanding the biochemistry of interactions between lipid antigens and CD1. The diversity of lipid antigens loaded onto CD1 molecules in normal cells, the ligand requirements for CD1 recognition by autoreactive T cells, including NK T cells, and the significance of CD1-mediated autoreactivity, as opposed to pathogen reactivity, remain controversial issues. PMID- 10375560 TI - Membrane compartmentation and the response to antigen. AB - Antigen-receptor engagement is a complex, highly orchestrated rearrangement of signaling molecules at, and adjacent to, the plasma membrane. Recent discoveries have shown that the plasma membrane itself is differentiated into microdomains of distinct lipid constituents and that the affiliation of lipid-modified proteins with those domains has important implications for antigen-receptor function. Disruption of lipid microdomains by a variety of methods attenuates TCR signal transduction. PMID- 10375561 TI - The role of B cells in acute and chronic infections. AB - The important role of B cells in protection against secondary viral infections has been recognized for a long time. Recent evidence suggests that B cells are also critically involved in protective immune reactions classically attributed to T cells. Specifically, antibodies have been documented to protect from many primary viral and parasitic infections and to be indispensable for the control of latent viral infections. Current vaccine strategies should take into account this pivotal role of antibodies. PMID- 10375562 TI - Seeing the world in red and blue: insight into plant vision and photoreceptors. AB - Plants see light through multiple photoreceptors, including phytochromes and cryptochromes. Cryptochromes are flavoproteins that participate in many blue light responses, including phototropism in plants and entrainment of circadian rhythms in plants and animals. A novel flavoprotein, NPH1, is also implicated in plant phototropism. Phytochromes function as serine/threonine kinases whose potential interacting partners include cryptochrome (CRY1 and CRY2). PMID- 10375563 TI - Making starch. AB - Improvements in understanding the structure of the starch granule and the nature and roles of starch-synthesising enzymes have allowed detailed mechanisms of the synthesis of the amylopectin and amylose components of the granule to be suggested. However, none of these proposed mechanisms has yet been shown to operate in vivo. Several critical aspects of granule synthesis, including granule initiation and the formation of the growth rings, remain a mystery. PMID- 10375564 TI - Biochemical dissection of photorespiration. AB - Progress has been made in the understanding of photorespiration and related proteins (Rubisco, glycolate oxidase and glycine decarboxylase) in the context of recent structural information. Numerous shuttles exist to support transamination, ammonia refixation and the supply or export of reductants generated or consumed (via malate-oxaloacetate shuttles) in the photorespiratory pathway. A porin-like channel that is anion selective represents the major permeability pathway of the peroxisomal membrane. PMID- 10375565 TI - Iron acquisition by plants. AB - In nongraminaceous plants, the FeII-transporter gene and ferric-chelate reductase gene have been cloned from Arabidopsis thaliana, whereas FeIII-reductase has not. In graminaceous monocots, the genes for mugineic acids (MAs) synthesis, nas (nicotianamine synthase) and naat (nicotianamine aminotransferase), have been cloned from barley, whereas the FeIII-MAs transporter gene is yet to be cloned. Transferrin absorption in Dunaliella has been reported, suggesting a phagocytotic (endocytotic) Fe-acquisition mechanism. Work to develop transgenic cultivars tolerant to Fe-deficiency in calcareous soils is now in progress. PMID- 10375566 TI - Redundancy as a way of life - IAA metabolism. AB - Plants have evolved elaborate systems for regulating cellular levels of indole-3 acetic acid (IAA). The redundancy of this network has complicated the elucidation of IAA metabolism, but molecular genetic studies and precise analytical methods have begun to expose the circuitry. It is now clear that plants synthesize, inactivate and catabolize IAA by multiple pathways, and multiple genes can encode a particular enzyme within a pathway. A number of these genes are now cloned, which greatly facilitates the future dissection of IAA metabolism. PMID- 10375567 TI - Sugar transporters in plant biology. AB - Sugar transporters are key players in many fundamental processes in plant growth and development. Recent results have identified several new transporters that contribute to a wide array of physiological activities, and detailed molecular analysis has provided exciting insights into the structure and regulation of these essential membrane proteins. PMID- 10375568 TI - Source-sink regulation by sugar and stress. AB - The regulation of carbon partitioning between source and sink tissues in higher plants is not only important for plant growth and development, but insight into the underlying regulatory mechanism is also a prerequisite to modulating assimilate partitioning in transgenic plants. Hexoses, as well as sucrose, have been recognised as important signal molecules in source-sink regulation. Components of the underlying signal transduction pathways have been identified and parallels, as well as distinct differences, to known pathways in yeast and animals have become apparent. There is accumulating evidence for crosstalk, modulation and integration between signalling pathways responding to phytohormones, phosphate, light, sugars, and biotic and abiotic stress-related stimuli. These complex interactions at the signal transduction levels and co ordinated regulation of gene expression seem to play a central role in source sink regulation. PMID- 10375569 TI - Nitrate regulation of metabolism and growth. AB - Recent research shows that signals derived from nitrate are involved in triggering widespread changes in gene expression, resulting in a reprogramming of nitrogen and carbon metabolism to facilitate the uptake and assimilation of nitrate, and to initiate accompanying changes in carbon metabolism. These nitrate derived signals interact with signals generated further downstream in nitrogen metabolism, and in carbon metabolism. Signals derived from internal and external nitrate also adjust root growth and architecture to the physiological state of the plant, and the distribution of nitrate in the environment. PMID- 10375570 TI - Zeroing in on zinc uptake in yeast and plants. AB - Zinc is an essential micronutrient. Genes responsible for zinc uptake have now been identified from yeast and plants. These genes belong to an extended family of cation transporters called the ZIP gene family. Zinc efflux genes that belong to another transporter family, the CDF family, have also been identified in yeast and Arabidopsis. It is clear that studies in yeast can greatly aid our understanding of zinc metabolism in plants. PMID- 10375571 TI - Plasma membrane transport in context - making sense out of complexity. AB - Major advances in our understanding of the transport of inorganic nutrient ions across plant plasma membranes have emerged from recent studies on the control of the dominant H+-pumping ATPase and from identification of a range of new transporters for divalent cations, potassium, phosphate and nitrate. In many cases, multiple transporter isoforms have been described. An appreciation of the physiological roles of these transporters demands combined genetic and physiological approaches, which, in the case of an outward rectifying K+ channel, have already been used to yield an intriguing insight into root-mediated K+ release into the xylem. In this review we attempt to place some of those developments in a physiological context. PMID- 10375572 TI - Connections between virus movement, macromolecular signaling and assimilate allocation. AB - Studies originating with plant viruses led to the concept that plasmodesmata potentiate the cell-to-cell trafficking of viral and endogenous proteins and nucleoprotein complexes. In this article, we develop the theme that, at the tissue/organ level, cell-to-cell trafficking of information molecules enables non cell-autonomous control over a range of processes, whereas at the organismal level, the phloem serves as an information superhighway. The capacity to deliver proteins and nucleoprotein complexes, over long distances, allowed for the development of a viral surveillance/resistance mechanism, as well as the integration of processes at the whole-plant level. PMID- 10375586 TI - Dorothy Hodgkin: A Life. PMID- 10375588 TI - Cooperativity between free and N-(2-hydroxypropyl) methacrylamide copolymer bound adriamycin and meso-chlorin e6 monoethylene diamine induced photodynamic therapy in human epithelial ovarian carcinoma in vitro. AB - The purpose of this study was to determine the interaction between free (unbound) and N-(2-hydroxypropyl) methacrylamide (HPMA) copolymer bound adriamycin and meso chlorin e6 monoethylene diamine (Mce6) induced photodynamic therapy in combination in their cytotoxic activities against human ovarian epithelial carcinoma (OVCAR-3) in vitro. The effects of each agent (free drugs and HPMA copolymer bound) alone and in combination were measured simultaneously utilizing two measures of cell viability: a) mitochondrial respiration via the 3-(4,5 dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide reduction (MTT) assay; and b) thymidine incorporation via the tritiated thymidine incorporation (TI) assay. These were performed at 72 and 144 h after drug exposure. Forty-eight hours from time zero (24 h after drug addition), the cells treated with Mce6 (free and HPMA copolymer bound) and controls were exposed to 650 nm light (13 min at 15 mW/cm2, 11.7 J/cm2). The calculated ED50 values by the MTT 72 h assay for adriamycin (A) and Mce6/light (C) were 1.5 microg/ml and 209 ng/ml, respectively. Adriamycin demonstrated progressive cellular toxicity over time in both assays. Mce6/light demonstrated initial damage at 72 h by MTT and TI which recovered by 144 h. Adriamycin and Mce6/light acted cooperatively to increase the percentage of cells inhibited. In combination, 21.3+/-1.5% MTT reduction activity was observed by free adriamycin and Mce6/light compared to the expected 27+/-5% (p<0. 0001) based on additivity. Twice the ED50 of adriamycin (2A=3 microg/ml) or Mce6/light (2C=418 ng/ml) resulted in only 42+/-3.6% and 39.2+/-2.0% activity, respectively (both p<0.0001 vs. combination). When Mce6/light at 10x ED50 (10C) was combined with 1x ED50 of adriamycin (1A), or the reciprocal combination, additional cooperativity was demonstrated. Compared to free drugs, both HPMA copolymer bound adriamycin (P-A) and HPMA copolymer bound Mce6/light (P-C) required a 10-fold increase in drug concentration to show equivalency with free drugs (A or C). Dose response curves demonstrated a reduced slope compared to free drugs in the same dose ranges. When P-A was added (1-10x free adriamycin ED50) to an effective concentration of P-C (10P-C: equivalent to 10x free Mce6 ED50) an improved long-term inhibition of OVCAR-3 cell multiplication was noted in both the MTT and TI 144 h assays. P-C (1-10x free Mce6 ED50) added to an effective concentration of P-A (10P-A: equivalent to 10x free adriamycin ED50) did not appear to significantly improve the efficacy profile of P-A. A and C in vitro appear to act independently and are cooperative in their combined toxicity against the human ovarian epithelial carcinoma cell line OVCAR-3. HPMA copolymer adriamycin and Mce6 conjugates (P-A and P-C, respectively) inhibited growth of OVCAR-3 in vitro. HPMA copolymer-adriamycin added to HPMA copolymer-Mce6 improved the efficacy of HPMA copolymer-Mce6. PMID- 10375589 TI - Involvement of the Ink4a gene (p16 and p19arf) in murine tumorigenesis. AB - The INK4A and INK4B genes map to 9p21, with the INK4A gene encoding two products, p16 and p19ARF. Many neoplasms in which INK4A and INK4B genes are altered show deletions involving both genes. Mice carrying a targeted Ink4a deletion develop tumors at an early age. In the present study we examined the genetic alterations affecting the remaining Ink4a allele and the Ink4b gene in tumors arising in heterozygous Ink4a mice. We identified deletion of the remaining Ink4a allele in 7 of 18 (39%) tumors. We also observed deletion of the exon 1beta in 3 cases, one of them presenting this deletion as a unique alteration. In conclusion, the deletion of the remaining Ink4a allele was the alteration most frequently observed, representing the inactivation of two proteins capable of arresting the cell cycle through different pathways that involve the tumor suppressors pRB and p53. PMID- 10375590 TI - Sentinel node localisation: A new prospective in the treatment of nodal melanoma metastases. AB - Sentinel node (SN) mapping and biopsy is a procedure that accurately stages the regional lymph node (LN) basin. Defined patterns of lymphatic drainage allow intraoperative determination of the first (sentinel) lymph node in the regional basin, and the absence of metastatic disease in the SN accurately reflects the absence of melanoma in the remaining regional nodes. The use of radiocolloid and a hand-held gamma detecting probe (GDP) together with a vital blue dye provides optimal results, and allows for the successful identification of the SN in over 99% of the procedures. Close collaboration between surgeons, nuclear radiologists and pathologists is required to ensure optimal results. Examination of serially sectioned SNs by hematoxylin-eosin staining (H&E), immunohistochemical staining and perhaps in the near future RT-PCR should reduce the number of patients with missed microscopic melanoma in the regional lymph nodes. Furthermore, the survival benefit recently reported in patients with melanoma metastatic to regional nodes using high dose of interferon alpha-2b signals that the surgeons should aggressively examine patients for the presence of occult regional melanoma metastases. Intraoperative SN mapping and SN biopsy are cost-effective procedures that allows accurate identification of regional lymph nodes that contain metastatic melanoma. PMID- 10375591 TI - Activated lansoprazole inhibits cancer cell adhesion to extracellular matrix components. AB - Integrins play an important role in tumor dissemination. The purpose of this study was to examine whether a SH-reactive reagent inhibits the adhesion of the human pancreatic cancer cell line AsPC-1 to extracellular matrix components. Activated lansoprazole (AG-2000) was used as the SH-reactive reagent because this compound is known to react with SH groups but does not permeate the cell membrane. The effect of AG-2000 on the adhesion of AsPC-1 cells to matrix was examined, using both an in vitro adhesion assay and an in vivo nude mouse xenograft model of peritoneal implantation. In the in vitro adhesion assay, a 60 min exposure of AsPC-1 cells to AG-2000 resulted in a dose-dependent inhibition of AsPC-1 cell adhesion to laminin, fibronectin and type IV collagen, although AG 2000 did not affect the viability of AsPC-1 cells by MTT assay. In the in vivo assessment of AsPC-1 cell implantation, the AsPC-1 cells were initially preincubated with AG-2000 for 60 min to ensure adequate exposure of the AsPC-1 cells to AG-2000 before intraperitoneal injection. AG-2000 significantly inhibited the peritoneal implantation of the AsPC-1 cells in nude mice. These findings suggest that a short exposure of cancer cells to AG-2000 can inhibit cancer cell adhesion to extracellular matrix components. PMID- 10375592 TI - Cytogenetic and CGH studies of four neuroendocrine tumors and tumor-derived cell lines of a patient with multiple endocrine neoplasia type 1. AB - A malignant insulinoma (LOHG-I), a carcinoid of the lung (LOHG-L), a parathyroid adenoma (LOHG-NSA), and a fibroma (LOHG-F) were obtained from a patient with multiple endocrine neoplasia type 1 (MEN1). Long-term cultures were established. Essential neurobiological properties of the cell lines were proven immunocytochemically and by electron microscopy. Molecular analysis of the germline DNA showed a 4 bp deletion in exon 3 of the MEN1 gene. Cytogenetic and CGH analyses of the tumors/tumor cell lines revealed diploidy and balanced and unbalanced structural aberrations different for each tumor. Chromosomes 6q21, 11q and 17q were most frequently involved in clonal structural aberrations. PMID- 10375593 TI - Apoptosis in relation to proliferating cell nuclear antigen and Dukes' stage in colorectal adenocarcinoma. AB - Colorectal cancer is a disease that is associated with default in the balance of apoptotic regulation. In the present study apoptosis was examined in 158 colorectal adenocarcinomas using the terminal deoxynucleotidyl transferase mediated digoxigenin nick end labeling (TUNEL) method. The median apoptotic index (AI) was 0.95% (range 0-6. 68%). Eighty-two tumours exhibited AI 0.95%. We revealed a positive correlation between apoptosis and proliferation determined as the expression of proliferating cell nuclear antigen (PCNA, p=0.002). The frequency of apoptosis increased from Dukes' stage A, B, C to D (p=0.01). No correlations were found between apoptosis and the patients' sex, age, tumour location, growth pattern, differentiation, prognosis, bcl-2, p53 or K-ras. Our findings suggest that we should further investigate the relationship between apoptosis and cellular proliferative activity in colorectal cancer to evaluate whether this might provide additional information in the selection of patients for effective adjuvant therapy. PMID- 10375594 TI - A novel hydrophobized GM3 ganglioside/Neisseria meningitidis outer-membrane protein complex vaccine induces tumor protection in B16 murine melanoma. AB - Gangliosides are sialic acid-containing glycosphingolipids that have increased surface membrane expression on cancers of neuroectodermal origin. The present study was designed to investigate at a preclinical level the therapeutic usefulness of a consistently immunogenic and safe conjugate vaccine in melanoma. We have examined a novel vaccine of GM3 monosialoganglioside hydrophobically conjugated with the outer-membrane-protein complex from Neisseria meningitidis plus Montanide ISA 51 in the B16 melanoma mouse model. B16 cell line is characterized by the predominant presence of ganglioside GM3 on the cell surface. Vaccines were administered i.m. in the quadriceps at 14-day intervals and B16 cells were injected in the subcutis of the right flank of C57BL/6 mice, 7 days after the fourth dose. Significant suppression of tumor growth and prolongation of survival were seen by immunization with GM3 vaccine in animals challenged with 5x10(3) or 10(3) live melanoma cells. In addition, vaccination reduced tumor growth in animals challenged with 5x10(4) cells. The reactivity of serum IgG from vaccinated mice was examined by a sensitive immunoperoxidase assay on B16 tumor specimens. Most melanoma cells displayed a distinct positive staining associated with both cell membrane and cytoplasm. In accordance with the immunohistochemical stainings, the antisera of immunized mice reacted brightly against B16 melanoma cells in flow cytometry studies. Anti-sera also mediated complement-mediated cytotoxicity and specific response could be totally ascribed to antibodies of the IgG2b subclass. The present data suggest that GM3 vaccine may provide a useful immunotherapeutic strategy for melanoma. PMID- 10375595 TI - Decreased expression of galectin-3 in basal cell carcinoma of the skin. AB - Galectins are beta-galactoside-binding lectins that play multiple roles during tumor progression. Previous work conducted in our laboratory has demonstrated decreased galectin-3 expression in carcinomas from colon, breast, ovary and endometrium, compared to the corresponding normal tissues. In this study, we examined the pattern of galectin-3 expression by immunohistochemistry in a group of 10 basal cell carcinomas of the skin. In the surrounding normal skin, galectin 3 immunostaining was found predominantly in the middle epidermis (spine layer) and eccrine sweat glands. Compared to the normal epidermal cells, basal carcinoma cells observed in all 10 samples examined presented with significantly decreased galectin-3 immunostaining. These data further demonstrates that galectin-3 is down-regulated in a variety of human cancers, including basal cell carcinoma. PMID- 10375596 TI - Immunological and molecular characterization of an aggressive murine lymphoma variant: modulation in vitro and in vivo. AB - The highly metastatic murine ESb-L lymphoma was analyzed with respect to its possible origin and phenotype modulation. By determining the methylation status of the CD8 gene an early thymic origin of the ESb-L lymphoma cells is suggested. It revealed that the precursors of ESb-L cells had at least one CD8 allele expressed during their development. ESb-L tumor cells were found to express ICAM 1, ICAM-2, VLA-4 and Mel14 as adhesion molecules and homing receptors and CD25, CD69 and CD124 (HSA) as T-cell related activation markers. PCR analysis revealed that ESb-L tumor cells express a Th2-like cytokine pattern with mRNAs for IL-4, IL-5, IL-6, IL-10 and IL-13, but not for IL-2 and IFNgamma. In addition mRNA for TNFalpha, LT, IFNalpha and the chemokines MIP1alpha and MIP1beta was found. The expression of the adhesion molecules ICAM-1, ICAM-2, VLA-4 and of the T-cell activation marker CD25 on ESb-L tumor cells could be upregulated by incubating the cells with 10 ng/ml TNFalpha. For CD25 this effect was confirmed also at the mRNA level. Using the lacZ transduced T-cell lymphoma ESb-L-CI we were able to re isolate live tumor cells from the primary site or from a metastasized liver and to investigate their phenotype ex vivo. MIP1alpha mRNA expression was strongly reduced in ex vivo isolated tumor cells as compared to in vitro grown cells indicating the modulatory role of the tumor microenvironment. The presented data suggest possible roles of TNFalpha and/or other microenvironmental factors modulating the expression of molecules involved in cell migration and adhesion thereby influencing cancer metastasis. PMID- 10375597 TI - Alternative splicing, but not allelic loss, of the FHIT gene increases with development of lung cancer. AB - The FHIT gene is considered to be a tumor suppressor gene, its role and inactivation mechanism remain unclear. We analyzed FHIT gene aberrations in 64 lung cancer tissues and found that the appearance of the aberrant FHIT transcripts depends on the condition of RT-PCR and high telomerase activity, shortened telomere length, and advanced pathological stage were likely associated with the prevalence of aberrant FHIT transcripts, but not with allelic loss of the FHIT gene. These observations would indicate that an additional unknown gene may exist, which is more responsible for the allelic loss around the FHIT gene locus. PMID- 10375598 TI - Electrochemotherapy against colorectal carcinoma: comparison of in vitro cytotoxicity of 5-fluorouracil, cisplatin and bleomycin. AB - Effectiveness of electrochemotherapy against colorectal carcinoma (CRC) was investigated in vitro using murine CRC cell lines. Electropermeabilization did not increase the sensitivity of CRC cells to 5-fluorouracil or cisplatin. Conversely, electropermeabilization markedly increased the sensitivity of CRC cells to bleomycin (BLM), resulting in 1,000-fold higher susceptibility. Subsequent analyses revealed that electropermeabilization significantly increased intracellular BLM levels of CRC cells. These results suggest that electrochemotherapy with BLM is a promising modality for the treatment of CRC, and that electrochemotherapy can be translated from the treatment of superficial tumors to the treatment of internal tumors including CRC. PMID- 10375599 TI - Cepharanthin enhances thermosensitivity without a resultant reduction in the thermotolerance of a murine mammary carcinoma. AB - Cepharanthin (Ce) is a biscoclaurine alkaloid extracted from Stephania cepharantha Hayata. The results of our previous in vitro study indicated that Ce reduces thermotolerance by enhancing thermosensitivity. In the present study, we investigated the in vitro and in vivo effects of Ce on thermosensitivity and thermotolerance using a murine mammary carcinoma, MCa, and C3H/HeN mice. Ce enhanced the thermosensitivity of MCa cells for heating at 44 degrees C not only in vitro but also in vivo. The in vivo enhancement ratio +/- SD of Ce at 100 mg/kg for heating at 44 degrees C was 1.3+/-0.3. The fractionated heat treatments at 44 degrees C for 30 and 60 min with an interval time of 0-6 days resulted in the development of remarkable thermotolerance and the expression of heat shock protein 70 in MCa tumors after the first heating. Ce at 100 mg/kg given immediately after the first heating increased the expression of heat shock protein 70 in MCa tumors, and did not reduce the development of thermotolerance. Ce given immediately before the first or second heating also did not inhibit the thermotolerance. The results of this study suggest that Ce enhances the thermosensitivity of MCa tumors as a thermosensitizer, but that this mild thermosensitizing property of Ce might be insufficient to conquer the remarkable thermotolerance in MCa tumors that develops after the first heating. PMID- 10375600 TI - Suppression of tumorigenicity in cervical carcinoma HeLa cells by an episomal form of adeno-associated virus. AB - To compare the oncosuppressive activity of integrated and episomal adeno associated virus (AAV), two Epstein-Barr virus (EBV) derived episomal form vectors, p205 and p220, were used to generate plasmids containing episomal AAV. HeLa cells transfected with p205, p220, and the plasmids with forward (pA205-1, pA220-1) and reverse (pA205-1, pA220-2) orientation of inserted AAV DNA were designated H205, H220, HA205-1, HA220-1, HA205-1, and Ha220-2, respectively. The respective average copy numbers of pA205 and pA220 per cell are 4.9-6.9 and 4.3. The AAV-transfected HeLa cells displayed growth inhibition when compared to parental and the vector-transfected HeLa cells. Nude mice assay showed that the tumor size from HA205-1 and HA205-2 cells were 11.3 and 22.6%, and those from HA220-1 and HA220-2 cells were 54.8 and 57.3%, respectively, when compared to parental HeLa cells. HA205-1 cells containing forward AAV insert exerted more oncosuppressive effect than HA205-2 cells containing reverse AAV insert. Our data indicate that the episomal AAV can exert oncosuppressive activity as compared to the integrated AAV in HeLa cells. PMID- 10375601 TI - In vivo gene transfer of a suicide gene under the transcriptional control of the carcinoembryonic antigen promoter results in bone marrow transduction but can avoid bone marrow suppression. AB - We constructed the CEA419/CD retrovirus vector carrying the cytosine deaminase (CD) gene directed by the carcinoembryonic antigen (CEA) promoter. pCD2 retrovirus vector carrying the CD gene directed by the retrovirus long terminal repeat promoter was also used. When mice bearing intraperitoneally disseminated colorectal carcinomas (CRCs) were infused intraperitoneally with pCD2 or CEA419/CD retrovirus-producing cells, a CD fragment was detected in CRCs and bone marrow cells. It was shown that the CD gene was expressed both in CRCs and in the bone marrow of animals infused with pCD2 retrovirus-producing cells, while the CD gene was expressed solely in CRCs of animals infused with CEA419/CD retrovirus producing cells. These results indicate that the use of a tumor-selective promoter may warrant the safety of in vivo gene therapy using suicide genes. PMID- 10375602 TI - Use of cellular telephones and the risk for brain tumours: A case-control study. AB - The use of cellular telephones has increased dramatically during the 1990's in the world. In the 1980's the analogue NMT system was used whereas the digital GSM system was introduced in early 1990's and is now the preferred system. Case reports of brain tumours in users initiated this case-control study on brain tumours and use of cellular telephones. Also other exposures were assessed. All cases, both males and females, with histopathologically verified brain tumour living in Uppsala-Orebro region (1994-96) and Stockholm region (1995-96) aged 20 80 at the time of diagnosis and alive at start of the study were included, 233 in total. Two controls to each case were selected from the Swedish Population Register matched for sex, age and study region. Exposure was assessed by questionnaires supplemented over the phone. The analyses were based on answers from 209 (90%) cases and 425 (91%) controls. Use of cellular telephone gave odds ratio (OR) = 0.98 with 95% confidence interval (CI) = 0. 69-1.41. For the digital GSM system OR = 0.97, CI = 0.61-1.56 and for the analogue NMT system OR = 0.94, CI = 0.62-1.44 were calculated. Dose-response analysis and using different tumour induction periods gave similar results. Non-significantly increased risk was found for tumour in the temporal or occipital lobe on the same side as a cellular phone had been used, right side OR = 2.45, CI = 0.78-7.76, left side OR = 2.40, CI = 0.52-10.9 Increased risk was found only for use of the NMT system. For GSM use the observation time is still too short for definite conclusions. An increased risk for brain tumour in the anatomical area close to the use of a cellular telephone should be especially studied in the future. PMID- 10375603 TI - A protective role for common p21WAF1/Cip1 polymorphisms in human ovarian cancer. AB - The objective of this study was to compare the frequency of two common p21WAF1/Cip1 gene polymorphisms in ovarian cancer patients with that in age matched controls, from a population originating from Eastern Scotland. Both polymorphisms were found significantly less frequently in both the constitutive and tumour tissue DNA of ovarian cancer patients (3/65; 4.6%), than in that from geographically and age-matched controls (25/186; 13.4%) (p=0.0495, chi2). Furthermore, we found no p21WAF1/Cip1 gene mutations in any of the tumours, reflected by a relatively low degree of loss of heterozygosity (LOH) at the chromosomal region where the gene maps, providing further evidence that the p21WAF1/Cip1 gene is not mutated in ovarian cancer. The data suggest however, that there may potentially be a protective function for the two p21WAF1/Cip1 gene polymorphisms in the population under study. PMID- 10375604 TI - Tumor necrosis factor and gamma-interferon repress transcription from the c-myc P2 promoter by reducing E2F binding activity. AB - Transcription of the c-myc gene in HeLa cells has been shown to be repressed by the combined action of tumor necrosis factor (TNF) and gamma-interferon (gamma INF). Shown here, these two cytokines inhibit proliferation of Hela cells with a coordinate inhibition of c-myc gene expression. It was found that these two cytokines exert their effects on the more proximal region of the c-myc P2 promoter. Using c-myc promoter:CAT constructs, it was found that the combined action of these cytokines significantly repress transcription from P2. This repression occurred through the E2F site within the promoter and not the ME1a2 or ME1a1 sites. However, these cytokines had no effect on transcription from the rous sarcoma virus promoter or the SV40 virus early promoter. Protein binding assays indicate that TNF and gamma-INF did not effect the ability of the ME1a2 factor to bind to its site but did significantly repress E2F factor binding to its DNA sequence. PMID- 10375605 TI - Overexpression of protein kinase C-alpha in MCF-7 breast cancer cells results in differential regulation and expression of alphavbeta3 and alphavbeta5. AB - MCF-7 breast cancer cells stably transfected with protein kinase C-alpha (MCF-7 PKC-alpha cells) show anchorage-independent growth and exhibit increased tumorigenicity in nude mice. Since integrins are involved in tumor growth and metastatic spread, we investigated whether integrin expression is differentially regulated in MCF-7-PKC-alpha cells. Fluorescence-activated cell sorting revealed that alphavbeta3 is highly expressed on MCF-7-PKC-alpha cells, but is undetectable on MCF-7V cells (MCF-7 cells transfected with vector only). In contrast, MCF-7-PKC-alpha cells have reduced expression of alphavbeta5. Blocking experiments with antibodies to alphavbeta3 and alphavbeta5 revealed that these receptors are used by MCF-7-PKC-alpha cells to adhere primarily to vitronectin and osteopontin. Consistent with heterodimer expression, MCF-7-PKC-alpha cells express increased beta3 and decreased beta5 on their surface. Surface expression of alphav on MCF-7-PKC-alpha cells is unchanged. Western blotting, Northern analysis, and nuclear run-on assays indicated that post-translational mechanisms increase the surface expression of beta3 on MCF-7-PKC-alpha cells. In contrast, reduced beta5 transcription diminishes beta5 surface expression on MCF-7-PKC alpha cells. These results indicate that overexpression of PKC-alpha in MCF-7 cells alters beta5 and beta3 expression by transcriptional and post-translational mechanisms, respectively, resulting in altered heterodimer expression. These findings suggest that the increased metastatic capacity of tumor cells with elevated protein kinase C levels may result, in part, from modulation of integrin expression. PMID- 10375606 TI - Analysis of Bcl-2, Bax and Survivin genes in uterine cancer. AB - To investigate the role of the apoptosis-related genes, Bcl-2, Bax and Survivin genes were analyzed. For the Bax gene, abnormality was detected in 1 of 7 cervical and 1 of 6 endometrial cancer cell lines, 1 of 25 cervical cancer tissues and none of 17 endometrial cancer tissues using PCR-SSCP. In 4 cervical and 2 endometrial cancer cell lines, the ratio of Bcl-2 to Bax expression was higher than the control ratio using Western blotting. Survivin mRNA was detectable in all cell lines and all cancer tissues. The data suggested that these apoptosis-related genes may play important roles in the pathway of carcinogenesis of human uterine cancer. PMID- 10375607 TI - Involvement of galectin-3 expression in colorectal cancer progression and metastasis. AB - Galectin-3 is a beta-galactoside-specific lectin that binds to laminin sugar sites and is involved in tumor malignancy. Galectin-3 expression in relation to primary tumor and liver metastasis of colorectal cancer was examined to determined its involvement in cancer progression and metastasis. Immunohistochemical staining of galectin-3 was performed on 117 primary lesions and 15 liver metastases of colorectal cancer using TIB166 monoclonal antibody. The expression of galectin-3 was evaluated by grading the intensity of the staining as either negative, weakly positive, or strongly positive. Normal mucosa of all patients were strongly positive for galectin-3, but the staining in these tissues was still significantly less than in the primary lesions of the cancer (31.6%). Galectin-3 expression in the primary lesions was significantly increased, correlating with the progression of clinical stage (p=0. 0224), liver metastasis (p<0.0001), venous invasion (p=0.0048), and lymph node metastasis (p=0.0289). Liver metastatic lesions also showed up-regulated levels of galectin 3 compared to the primary lesions (p=0.0030). The group showing strongly positive galectin-3 had a significantly poorer prognosis than the negative/weakly positive group in terms of disease-free survival (p=0.0224). The strong expression of galectin-3 in colorectal cancer correlates with cancer progression, liver metastasis, and poor prognosis for patients. PMID- 10375608 TI - Angiogenesis and platelet-derived endothelial cell growth factor/thymidine phosphorylase expression in cervical cancer. AB - The object of this study was to clarify the association of platelet-derived endothelial cell growth factor (PD-ECGF)/thymidine phosphorylase (dThdPase), separately assessed in cancer cells and in stroma cells, with clinicopathological factors including tumor angiogenesis and prognosis in cervical cancer. The expression of PD-ECGF was evaluated by immunohistochemical staining in 92 patients with stage Ib-II cervical cancer. The microvessel count was assessed by immunostaining for factor VIII-related antigen in the most neovascularized area. Microvessel count was significantly higher in tumors with non-squamous cell carcinoma. PD-ECGF expression in cancer cells was significantly higher in tumors with pelvic node metastasis and squamous cell carcinoma. Immunopositivity for PD ECGF in stroma cells was significantly higher in tumors with large size and deep stromal invasion. The microvessel counts in cases with positive PD-ECGF expression in stroma cells were significantly higher than those in cases with negative PD-ECGF expression in stroma cells (p=0.048). Disease-free survival and overall survival were significantly worse in patients with deep stromal invasion, parametrial involvement, vaginal involvement, lymph-vascular space involvement, pelvic lymph node metastasis and high microvessel count. A multivariate analysis using Cox's proportional hazard model showed that high microvessel count independently predicted disease-free and overall survival. The expression of PD ECGF in stroma cells may play a crucial role in the promotion of angiogenesis and tumor angiogenesis can be used as a useful prognostic marker for cervical cancer. PMID- 10375609 TI - A comparison of the modulation of antiblastics cytotoxicity by verapamil and dipyridamole in a human colon carcinoma cell line. AB - This study was designed to compare the activity of two MDR modulators, verapamil and dipyridamole, on the in vitro growth of a human colon carcinoma cell line. The aims were: a) to investigate the different sensitivity of the parental cell line (LoVo S) and the doxorubicin-resistant one (LoVo R) towards the treatment with several antiblastics and their associations with verapamil or dipyridamole; b) to evaluate if the combined use of these drugs with verapamil or dipyridamole increases their cytotoxicity; c) to understand whether the mechanism of action of each modulator is the same. Idarubicin and vinblastine were the most active drugs on both cell lines. LoVo R cells showed cross-resistance to vinblastine, teniposide and mitoxantrone, while chemosensitivity towards cisplatin and cyclophosphamide was almost the same in both cell lines. The inhibitory effect on cell growth was enhanced when the drugs were associated with verapamil, but no difference was detected with cisplatin and cyclophosphamide. Verapamil is thus an effective MDR modulator when used with drugs actively pumped out of tumour cells by P-glycoprotein, while it is ineffective with drugs that induce resistance by different mechanisms. When combined with dipyridamole, a significant result was observed in the case of cisplatin, where a marked increase of cytotoxicity was detected. PMID- 10375610 TI - Induction of differentiation accompanies inhibition of Cdk2 in a non-small cell lung cancer cell line. AB - Induction of differentiation in a variety of model systems is accompanied by cell cycle exit and inhibition of Cdk2 kinase activity. We asked whether inhibition of Cdk2 activity is sufficient to allow differentiation to occur in a non-small cell lung cancer cell line. Treatment of NCI-H358 with flavopiridol, an inhibitor of multiple Cdk's, resulted in growth arrest and induction of mucinous differentiation. The onset of differentiation coincided temporally with loss of Cdk2 kinase activity. Western analysis revealed that flavopiridol treatment resulted in depletion of both cyclin E and D1, suggesting that loss of the regulatory subunits is at least partially responsible for the loss of Cdk kinase activity. Similarly, roscovitine, an inhibitor of Cdk's 1, 2, and 5, but not Cdk4, also induced differentiation in NCI-H358, although the resulting pattern of expression of cell cycle regulatory genes differed from the pattern obtained with flavopiridol. Furthermore, stable expression of an antisense Cdk2 construct in NCI-H358 also resulted in the appearance of a marker of mucinous differentiation. These results show that the inhibition of activity of cyclin dependent kinases, particularly Cdk2, by multiple different mechanisms is accompanied by differentiation. Thus, induction of differentiation is one potential mechanism of action for agents that down-regulate Cdk activity. PMID- 10375611 TI - Elevated blood levels of soluble tumor necrosis factor receptors in nasopharyngeal carcinoma: correlation with humoral immune response to lytic replication of Epstein-Barr virus. AB - Nasopharyngeal carcinoma (NPC) is tightly associated with Epstein-Barr virus (EBV) infection and a heavy infiltration of lymphoid cells in the tumor tissue. Although various lines of evidence have shown that the immune systems of NPC patients have the potential to attack the tumor cells, it is not yet understood how this potential is blocked. In this study we determined the circulatory soluble tumor necrosis factor receptors (sTNFRI and sTNFRII), which are proven to be inhibitory to the anti-tumor effects of tumor necrosis factor-alpha (TNF alpha), in NPC patients. The serum concentration of both sTNFRI and sTNFRII was determined with an ELISA method, and shown to be significantly higher in 28 NPC patients than in matched healthy controls. This elevation was found to be positively correlated with the serum titers of IgA against EBV early antigens and viral capsid antigens in NPC patients, suggesting that the increased serum concentration of sTNFRI and sTNFRII is possibly due to the EBV infection in NPC tumor cells. This is partly supported by FACS analysis of the circulatory T cells. Phenotypical expression of activation markers such as CD25, CD38, CD69 and CD71 in blood T cells was not significantly different between the NPC and control individuals, indicating the elevation of the sTNFRs is indeed derived from the local immune response in the tumor area. Based on these results, it seems that the increased sTNFRs may act as an inhibitor to decrease the host immune response towards tumor cells in NPC patients. PMID- 10375612 TI - Identification of c-myc promoter-binding protein and X-box binding protein 1 as interleukin-6 target genes in human multiple myeloma cells. AB - Interleukin-6 (IL-6) is implicated in the in vivo proliferation of malignant plasma cells in multiple myeloma. To define the molecular basis of the IL-6 induced mitogenic response in myeloma cells, we applied STAR (subtractive transcriptional amplification of mRNA), a new differential expression analysis technology, to isolate mRNAs preferentially expressed in IL-6-treated versus untreated cultures of the factor-responsive myeloma cell line U266. From the resulting collection of STAR clones, sequence information was obtained for a total of 72 distinct transcripts. Of these, 29 were found to correspond to known genes, 22 matched expressed sequence tags in public databases and 21 showed no sequence similarity to any existing entries. Among the known genes uncovered in the screen were those encoding proteins that function in cell division, cell signalling and gene/protein expression. Northern blot analysis documented that two transcription factor genes chosen for further study, c-myc promoter-binding protein (MBP-1) and X-box binding protein 1 (XBP-1), were up-regulated in U266 cells about 3-fold relative to the cell cycle-dependent beta-actin gene 12 h after IL-6 treatment. Both genes were also similarly up-regulated by IL-6 in factor-dependent ANBL-6 myeloma cells. These results indicate that MBP-1 and XBP 1 are IL-6 genes in myeloma cells; as such, they may play a role in IL-6-mediated growth control in multiple myeloma. PMID- 10375613 TI - Prognostic factors in non-metastatic limb osteosarcoma: A 20-year experience of one center. AB - The purpose of this study was to evaluate the prognostic significance of variables in osteosarcoma. We performed a retrospective analysis of 35 patients with non-metastatic limb osteosarcoma that were treated between 1973 and 1994. The following variables were evaluated: age, sex, ethnic group, tumor histology and primary site, alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) levels at diagnosis, treatment regimen, and the histologic response to treatment. Three variables showed significant correlation with prognosis: i) histologic response to preoperative treatment. Disease-free survival (DFS) was 89% in patients with grade III-IV histologic response after a median follow-up (MFU) of 64 months, 67% in patients with grade II after an MFU of 64 months, the patients with grade I response died within 15 months (p<0.0001); ii) treatment regimen. DFS was 83% after an MFU of 42 months, 62% after an MFU of 82 months, and 30% after an MFU of 177 months in patients treated by the 90's, 80's, and 70's protocols, respectively (p<0.05); iii) corrected ALP (cALP) levels at diagnosis. DFS was 78% after an MFU of 88 months in patients with cALP levels <200, and 32% after an MFU of 56 months in patients with cALP levels >200 (p=0.01). Low ALP levels, good histologic response to preoperative chemotherapy, and the new therapeutic regimen correlated with good prognosis in patients with osteosarcoma. PMID- 10375614 TI - Expression and secretion of TGF-beta isoforms and expression of TGF-beta receptors I, II and III in normal and neoplastic human breast. AB - We investigated gene expression of the TGF-beta signalling system (including peptides and receptors) in normal and malignant breast tissue. Additionally, gene and protein expression was determined in a series of primary epithelial and stromal cultures derived from these tissues. TGF-beta isoforms and their receptors were expressed by both tissue sets, however the percentage of samples expressing each transcript varied. In normal breast, both TGF-beta1 and TGF-beta3 were found in most samples (88 and 89% respectively), with fewer expressing TGF beta2 (68%). A similar pattern was evident in the tumours. Type I receptor of TGF beta was constitutively expressed in normal breast and observed in most tumours (90%). Type II and III receptors of TGF-beta were expressed less frequently, although the type II receptor was mainly expressed by tumours (P=0. 0075). All primary cultures produced TGF-beta1 and TGF-beta2. Comparing respective cell populations, tumour stromal cells produced significantly more TGF-beta1 than those derived from normal breast (P<0.0001). Linear regression analysis showed stromal cultures derived from breast tumours exhibited a strong positive correlation (r=0.976) in the production of TGF-beta1 and TGF-beta2. Thus, TGF beta and TGF-beta-receptors are widely and differentially expressed by normal and malignant breast and secretion of this peptide by epithelial and stromal cultures, in particular those derived from tumours, confirms its potential as an autocrine/paracrine regulator in breast cancer. PMID- 10375615 TI - Plant MAP kinase kinase kinases structure, classification and evolution. AB - The increasing number of reports describing plant MAP kinase signalling components reflects the cardinal role that MAP kinase pathways are likely to play during plant growth and development. Relationship and structural analyses of plant MAP kinase kinase kinase related cDNAs and genes established, on one hand, the PMEKKs, which may be distinguished into the alpha, beta, gamma, and zeta groups, and, on the other hand, the PRAFs that consist of the delta, eta and theta groups. Plant MAP3Ks are characterized by different primary structures, but conserved within a single group. A relationship analysis, which included animal, fungal and plant MAP3Ks, revealed a high degree of diversity among this biochemically established set of proteins, thus suggesting a range of biological functions. Four major families emerged, namely the MEKK/STE11, including the PMEKKs, the RAF, including the PRAFs, as well as the MLK and CDC7 families. These four families showed phylum-dependent distributions. Signature sequences characterizing the RAF family and the RAF subfamilies have been evidenced. However, no equivalent sequence motifs were identified for the MEKK/STE11 family, which is highly heterogeneous. PMID- 10375616 TI - Cloning of the cDNA encoding phenylalanyl tRNA synthetase regulatory alpha subunit-like protein whose expression is down-regulated during differentiation. AB - Hybrid polar compounds (HPCs), such as suberoylanilide hydroxamic acid (SAHA), induce differentiation of transformed cells. Differential display of RNA was used to identify genes whose expression is changed during SAHA-induced differentiation of murine erythroleukemia (MEL) cells. One such cDNA was identified whose mRNA level decreased by 50% after 8h of SAHA treatment as determined by Northern blot analysis. The full-length cDNA (1944bp in length) was cloned by sequencing of an EST clone and rapid amplification of 5' cDNA ends (5'-RACE). The predicted amino acid sequence is 589 amino acids and shares 45% identity with the yeast cytoplasmic phenylalanyl tRNA synthetase (PheRS) regulatory alpha-subunit. Human EST clones which share over 90% identity of predicted amino acid sequence with this cDNA map to chromosome 2 near the paired box homeotic gene 3 (PAX3) locus, a region syngenic to mouse chromosome 1. This is the first report of the cloning of the full-length cDNA for the murine PheRS regulatory alpha-subunit-like protein. The level of PheRS alpha-subunit-like mRNA is regulated during differentiation but not during cell cycle progression. PMID- 10375617 TI - Isolation and characterization of genomic sequences involved in the regulation of the human reduced folate carrier gene (RFC1). AB - Decreased reduced folate carrier (RFC) activity has been associated with MTX resistance in experimental models of transport-mediated MTX resistance, and has been attributed to changes in the expression of RFC1, the gene that encodes a protein with this activity. RNA transcripts of RFC1 may contain any one of four distinct 5' untranslated regions (UTRs). We cloned a human genomic DNA fragment upstream from the RFC1 translation start site and determined the nucleotide sequence of a 4.8kb region that contained the exons corresponding to each of the reported UTRs. To identify regulatory elements that may be involved in RFC1 transcription, we first developed a semi-quantitative RT-PCR assay using primers specific for each of the 5' UTRs to amplify RNA transcripts containing each of the RFC1 5' exons, and found evidence for differential transcription of RFC1 non coding exons in tissues, during development, and in MTX-resistant, transport deficient cells. We also found that RFC1 RNA levels are cell cycle regulated and peak at the G1 to S transition. Then, using a series of RFC1 promoter-reporter fusion constructs, we identified genomic sequences that may be involved in the regulation of expression of exons 1b and 1c of the RFC1 gene. These studies therefore identify regulatory regions of RFC1 promoters and potential models of regulation in which these regions may exert control. PMID- 10375618 TI - RXR-2, a member of the retinoid x receptor family in Schistosoma mansoni. AB - A cDNA encoding a second full-length member of the Schistosoma mansoni RXR family (SmRXR-2) was identified. The nucleotide sequence of SmRXR-2 translates into a protein of 784 amino acids with a pI of 7.63 and an approximate mass of 78kDa making it the largest reported RXR to date. Phylogenetic tree analysis provides evidence that SmRXR-2 is the most ancient full-length RXR identified. SmRXR-2 exhibits unique sequence features compared with other RXRs. RT-PCR results demonstrate that the SmRXR-2 gene is constitutively expressed and thus must play multiple roles throughout schistosome development in the vertebrate host. PMID- 10375619 TI - Characterization of the components of the putative mammalian sister chromatid cohesion complex. AB - Establishing and maintaining proper sister chromatid cohesion throughout the cell cycle are essential for maintaining genome integrity. To understand how sister chromatid cohesion occurs in mammals, we have cloned and characterized mouse orthologs of proteins known to be involved in sister chromatid cohesion in other organisms. The cDNAs for the mouse orthologs of SMC1S.c. and SMC3S.c. , mSMCB and mSMCD respectively, were cloned, and the corresponding transcripts and proteins were characterized. mSMCB and mSMCD are transcribed at similar levels in adult mouse tissues except in testis, which has an excess of mSMCD transcripts. The mSMCB and mSMCD proteins, as well as the PW29 protein, a mouse homolog of Mcd1pS.c./Rad21S.p., form a complex similar to cohesin in X. laevis. mSMCB, mSMCD and PW29 protein levels show no significant cell-cycle dependence. The bulk of the mSMCB, mSMCD and PW29 proteins undergo redistribution from the chromosome vicinity to the cytoplasm during prometaphase and back to the chromatin in telophase. This pattern of intracellular localization suggests a complex role for this group of SMC proteins in chromosome dynamics. The PW29 protein and PCNA, which have both been implicated in sister chromatid cohesion, do not colocalize, indicating that these proteins may not function in the same cohesion pathway. Overexpression of a PW29-GFP fusion protein in mouse fibroblasts leads to inhibition of proliferation, implicating this protein and its complex with SMC proteins in the control of mitotic cycle progression. PMID- 10375620 TI - Mutational analysis of the promoter region of the porA gene of Neisseria meningitidis. AB - The porA gene encodes the class 1 outer membrane protein (OMP1) in Neisseria meningitidis and is under transcriptional control. Promoter regions of porA from different clinical isolates were sequenced and were found to differ in the number of guanosine residues in a poly(G) track located upstream of the -10 region. Isolates that did not express OMP1 had up to nine G residues in the poly(G) track or an adenosine residue within this poly(G) track. Using beta-galactosidase as a reporter gene, the transcriptional activities of the promoter regions of the porA gene from three strains, two of which do not express OMP1, were assayed in both Escherichia coli and N. meningitidis. Mutations in the poly(G) track were created by site-directed mutagenesis and promoter fusions were further analyzed in E. coli and N. meningitidis. The number of nucleotides in the poly(G) track influenced promoter activity: reduction of a poly(G) track of 12nt by one and by two guanosine residues reduced promoter activity. Within the poly(G) track, replacement of an adenosine residue by a guanosine residue increased the promoter activity; replacement of a guanosine residue by an adenosine residue decreased the activity. The similar transcriptional activities for the mutated promoters in E. coli and N. meningitidis are compatible with similar control mechanisms for transcriptional control in both organisms. PMID- 10375621 TI - Metastatic rat carcinoma cells express a new retrotransposon. AB - Genes differentially expressed by a rat bladder carcinoma NBT-II cells and their in-vivo-selected metastatic M-NBT-II variant were analysed. Amplification and cloning of a 277-bp B sequence, exclusively expressed by the M-NBT-II cells, were performed, and this sequence was detected as a 6.7-kb RNA. This fragment shares 46-50% identities with the gag-related protein of mouse and hamster Intracisternal A Particles (IAPs). Screening of a M-NBT-II cDNA library with the B probe selected a 1671-bp sequence corresponding to the 5' end of a novel retrotransposon member of the rat IAP family. This sequence has a strong identity with the Ecker Rat IAP (ERA-IAP) except for the B portion and has an open reading frame potentially encoding a 114-amino-acid gag retrovirus-related protein. Rearrangement of this new retrotransposon could be relevant with the tumor progression in our model system since it is only expressed in the M-NBT-II in vivo-selected carcinoma metastasis. PMID- 10375622 TI - cDNA cloning of mouse tumor necrosis factor-alpha converting enzyme (TACE) and partial analysis of its promoter. AB - Tumor necrosis factor-alpha (TNFalpha) is a cytokine that induces pleiotropic inflammatory reactions. Soluble TNFalpha is released from its membrane-bound precursor by TNFalpha converting enzyme (TACE)/a disintegrin and metalloproteinase 17 (ADAM17). We have recently cloned the mouse TACE complete cDNA and the 5' flanking promoter region of the gene. Two versions of the mouse TACE cDNA having a coding region of 2541bp were obtained: one was about 4.1kb and the other was 4.4kb in length, in which only the length of the 3' UTR was different. Rapid amplification of 5' cDNA ends suggested that there were multiple transcriptional start sites. From the coding sequence, 827 amino acids were deduced which were 91.9 % identical to those of human TACE. Northern blot analysis indicated that the major transcript was approx. 4.4kb product in the mouse. The mouse TACE mRNA was ubiquitously expressed, and was particularly high in the lung. The proximal promoter contained multiple AP2 and Sp1 transcription factor binding sites and included a GC box and a CCAAT box, but lacked a consensus TATA box. Reporter gene analysis using RAW264.7 cells showed that the fragment at nt -290 to -1 from the translation start site has a strong promoter activity, and appeared to be essential for transcription of the mouse TACE mRNA. Finally, we found that the mouse TACE promoter at nt -2304 to -1 also worked in NIH3T3, 3T3L1 and C6 cell lines. PMID- 10375623 TI - The cDNA cloning and transient expression of an ovary-specific 17beta hydroxysteroid dehydrogenase of chickens. AB - A cDNA clone, pc17bHSD, was obtained from the chicken ovarian cDNA library by its partial homology to the cDNA sequence of the rat 17beta-hydroxysteroid dehydrogenase (17beta-HSD). The cDNA insert of pc17bHSD is 979bp long and contains an open reading frame (ORF) of 906bp. The deduced amino acid sequence of the ORF shows 48 and 50% overall identity with those of the rat and the human type-1 17beta-HSD, respectively. Five sequence regions common to the short-chain alcohol dehydrogenase superfamily are well conserved, including the YxxxK sequence motif at the active site. Northern blot hybridization detected a transcript of about 1kb only in ovaries of both sexually immature and mature female chickens. The 17beta-HSD activity, which was highly specific to the interconversion between estrone and estradiol-17beta, was detected in the cytoplasmic fraction of human 293 cells transfected transiently with an expression vector carrying the c17bHSD cDNA sequence, pcDNAI/c17bHSD. From these results, it is concluded that the pc17bHSD is the cDNA clone for the ovary specific molecular species of 17beta-HSD in chickens. PMID- 10375624 TI - The elongation factor-1delta (EF-1delta) originates from gene duplication of an EF-1beta ancestor and fusion with a protein-binding domain. AB - The molecular evolution of two components of elongation factor-1 (EF-1), EF-1beta and EF-1delta was analysed using the distance matrix, the maximum parsimony and the maximum likelihood methods, after careful alignment of protein and cDNA sequences. The topology of the phylogenetic trees obtained supports monophyly of plant EF-1beta and EF-1beta' sequences, and monophyly of higher eukaryotic animal EF-1beta and EF-1delta sequences. EF-1beta and EF-1delta are homologous in their C-terminal domain. EF-1delta, which emerged before arthropods, originates from a beta-type ancestor gene and fusion with a leucine zipper N-terminal motif. Plant EF-1beta and EF-1beta' correspond to paralogous genes whose ancestor was most likely duplicated before the emergence of monocotyledons and dicotyledons. PMID- 10375625 TI - The complete sequence of the mitochondrial genome of Daphnia pulex (Cladocera: Crustacea). AB - The sequence of the mitochondrial DNA (mtDNA) of the branchiopod crustacean Daphnia pulex has been completed. It is 15333bp with an A+T content of 62.3%, and contains the typical complement of 13 protein-coding, 22 transfer RNA (tRNA) and two ribosomal RNA (rRNA) genes. Comparison of this sequence with the sequences of the other eight completely sequenced arthropod mtDNAs showed that gene order and orientation are identical to that of Drosophila but different from Artemia due to the rearrangement of two tRNA genes. Nucleotide composition, codon usage, and amino acid composition are very similar in the crustaceans, but divergent from insects and chelicerates which show a much higher bias towards A+T. However, with few exceptions, the mitochondrial proteins of Daphnia are more similar to those of the dipteran insects (Drosophila and Anopheles) than to those of Artemia, at both the nucleotide and amino acid levels, suggesting that Artemia mtDNA is evolving at an accelerated rate. These results also show that sequence evolution and the evolution of nucleotide composition can be decoupled. Analysis of nucleotide substitution patterns in COII showed that there has been an unbiased acceleration of the overall substitution rate in Artemia. In contrast, the accelerated substitution rate in Apis is due partly to extreme A+T mutation pressure. Secondary structures are proposed for the Daphnia tRNAs and rRNAs. The tRNAs are similar to those of other arthropods but tend to have TPsiC arms that are only 4bp long. The rRNA secondary structures are similar to those proposed for insects except for the absence of a small number of helices in Daphnia. Phylogenetic analysis of second codon positions grouped Daphnia with Artemia, as expected, despite the latter's accelerated divergence rate. In contrast, the unusual pattern of mtDNA divergence in Apis led to a topology in which the holometabolous insects (Anopheles, Drosophila, Apis) appeared to be paraphyletic with respect to the hemimetabolous insect, Locusta, due to the early branching of Apis. PMID- 10375626 TI - Sequence analysis of the Mycoplasma arthritidis bacteriophage MAV1 genome identifies the putative virulence factor. AB - The bacteriophage MAV1 is required for the development of arthritis in rats after infection with its host Mycoplasma arthritidis. To identify the phage-encoded virulence factor for this arthritis, the complete nucleotide sequence of MAV1 was determined. The linear double-stranded genome of MAV1 is 15644bp and contains 15 ORFs. Putative protein products from these ORFs were identified by comparison of the deduced amino acid sequences to known proteins and comprise DNA replication, restriction-modification, structural, regulatory, and integration/excision proteins. Eight putative promoters were identified; four of these would produce polycistronic transcripts. Translation of each ORF appears to be initiated independently, with each having its own RBS. A single ORF, vir, was identified on the minus strand of the phage genome. The putative protein product of vir contains a classic prokaryotic lipoprotein signal sequence and is a strong candidate for the MAV1-encoded virulence determinant. PMID- 10375627 TI - The dnaK/dnaJ operon of Haemophilus ducreyi contains a unique combination of regulatory elements. AB - Haemophilus ducreyi, which causes the genital ulcer disease chancroid, requires high basal levels of the 60-kDa heat-shock (hs) protein GroEL in order to survive and adhere to host cells in the presence of common environmental stresses. In contrast, the 70-kDa hs protein, DnaK, a negative modulator of the hs response in prokaryotes, is not produced at as high a level as GroEL. Because of these differences, we were interested in identifying regulatory elements affecting the expression of the H. ducreyi dnaK/dnaJ operon. First, the genes encoding H. ducreyi DnaK (Hsp70) and DnaJ (Hsp40) were sequenced. The deduced amino acid sequences shared 82.8 and 63. 9% identity with the Escherichia coli DnaK and DnaJ homologs, respectively. Despite the presence of highly similar (but not identical) hs promoter sequences preceding both the H. ducreyi groES/groEL and dnaK/dnaJ operons, transcription levels for groEL were found to exceed that of dnaK. Subsequently, other genetic elements that could contribute to a lower basal expression of dnaK in H. ducreyi were identified. These elements include: (1) a complex promoter for dnaK consisting of four transcriptional start points (two for sigma32 and two for sigma70) identified by primer extension; (2) a putative binding site for Fur (a transcriptional repressor of iron-regulated genes) that overlaps the initiating AUG of dnaK; and (3) the potential for extensive secondary structure of the long leader sequences of the dnaK transcripts, which could interfere with efficient translation of DnaK. This unique combination of regulatory elements may be responsible for the relatively low-level expression of dnaK in this fastidious genital pathogen. PMID- 10375628 TI - Characterization of the functional replication origin of Mycobacterium tuberculosis. AB - The gene order in the 5kb Mycobacterium tuberculosis dnaA region is rnpA, rpmH, dnaA, dnaN and recF. We show that M. tuberculosis DNA fragment containing the dnaA-dnaN intergenic region functioned as oriC, i.e., allowed autonomous replication to otherwise nonreplicative plasmids, in M. tuberculosis H37Ra (H37Ra), avirulent strain of M. tuberculosis, and in Mycobacterium bovis BCG (BCG), a closely related, slowly growing mycobacterial strain. Removal of Escherichia coli plasmid replication origin (ColE1) from the M. tuberculosis oriC plasmids did not abolish their ability to function as oriC, confirming that the autonomous replication activity of these plasmids is due to the presence of the DNA fragment containing the dnaA-dnaN intergenic region. Deletion analyses revealed that the minimal oriC DNA fragment is 814bp. The copy number of M. tuberculosis oriC plasmids containing ColE1 ori relative to chromosomal oriC is one and the 5' flanking region of minimal oriC contains features that support stable autonomous replication. The M. tuberculosis oriC did not function in rapidly growing mycobacterial species such as M. smegmatis. M. smegmatis oriC functioned only in M. fortuitum, but not in any of the slowly growing mycobacterial species such as M. tuberculosis and BCG. Together these data suggest that the replication initiation mechanisms in the slowly growing Mycobacteria are similar and probably different from those in the rapidly growing Mycobacteria and vice versa. PMID- 10375629 TI - The micronuclear gene encoding a putative aminoacyl-tRNA synthetase cofactor of the ciliate Euplotes octocarinatus is interrupted by two sequences that are removed during macronuclear development. AB - The micronuclear gene of the ciliated protozoan Euplotes octocarinatus (Eo) syngen 1 encoding the putative aminoacyl-tRNA synthetase cofactor (ARCE), as well as its macronuclear version and the corresponding cDNA, were amplified and sequenced. Analyses of the sequences revealed that the micronuclear gene contains two sequences (430 and 625bp long) that are missing in the macronuclear version of this gene. These sequences are called 'internal eliminated sequences' (IESs) and appear to occur in all ciliates. The two IESs are located in the coding region of the micronuclear gene. One IES is flanked by a pair of dinucleotide 5' TA-3' direct repeats and the other one by a pair of hepta-nucleotide 5'-TTACTGA 3' direct repeats. Inside the two IESs, several other sequence repeats were found. The macronuclear DNA molecule carrying this gene is 1517bp long and shows characteristics typical of macronuclear chromosomes of hypotrichous ciliates. Copy number determination revealed that the molecule is amplified to only about 750 copies per macronucleus. The deduced protein is a 441-amino-acid (aa) polypeptide with a molecular mass of 50kDa. It shares a conserved endothelial monocyte-activating polypeptide II (EMAP II)-like carboxyl-terminal domain and a hydrophilic central domain containing a KEKE-motif with a group of proteins associated with aminoacyl-tRNA synthetases and tRNAs. PMID- 10375630 TI - Deletion of 24 open reading frames from chromosome XI from Saccharomyces cerevisiae and phenotypic analysis of the deletants. AB - As a part of the EUROFAN program, 24 open reading frames from Saccharomyces cerevisiae (YKR010c to YKR013w, YKR015c to YKR025w, YKR081c to YKR083c, YKR087c to YKR091w and YKR096w) were disrupted in two genetic backgrounds, FY1679 and W303. Systematic deletions and phenotypic analysis were performed following a hierarchical strategy, the so-called 'mass murder'. Of the 24 genes thus deleted, four are essential, whereas the deletion of 17 did not reveal any significant difference between the parental and mutant strains. Deletions of the remaining three show some growth phenotype; ykr024c mutants grow slowly under any conditions, ykr019c mutants grow slower in a rich medium and ykr082w mutants are temperature sensitive, being unable to germinate at 30 degrees C and above. PMID- 10375631 TI - Are horizontal transfers involved in the evolution of the Streptococcus thermophilus exopolysaccharide synthesis loci? AB - A 32.5kb variable locus of the Streptococcus thermophilus CNRZ368 chromosome, the eps locus, contains 25 ORF and seven insertion sequences (IS). The putative products of 17 ORF are related to proteins involved in the synthesis of polysaccharides in various bacteria. The two distal regions and a small central region of the eps locus are constant and present in all or almost all of the S. thermophilus strains tested. The other regions are variable and present in only some S. thermophilus strains tested, particularly in the closely related strains CNRZ368 and A054. A 13.6kb variable region of the eps locus of S. thermophilus CNRZ368 contains two ORF that are almost identical to epsL and orfY of the eps locus of Lactococcus lactis NIZOB40 and seven IS belonging to four different families, ISS1, IS981, IS1193 and IS1194. Five of these sequences were probably acquired by horizontal transfer from L. lactis (Bourgoin, F., et al., 1996. Gene 178, 15-23). Three probes of this 13.6kb region hybridized with the DNA of several L. lactis strains tested. A specific probe for another sequence within the S. thermophilus eps locus, epsF, hybridized with the DNA of one of the L. lactis strains tested. Sequence comparisons also suggest that five ORF of the eps locus have a mosaic structure and probably result from recombinations between sequences that are 10 to 50% divergent. The chimeric structure of the eps locus suggests a very complex evolution. This evolution probably involves both the acquisition of the 13.6kb region from L. lactis by horizontal transfer and exchanges within the S. thermophilus species. PMID- 10375632 TI - Molecular cloning of the mouse dopamine transporter and pharmacological comparison with the human homologue. AB - Drug abuse is a serious problem in the United States and in the world. Cocaine and amphetamines, widely used drugs of abuse, bind to dopamine (DA), serotonin, and norepinephrine transporters with high affinity and block their functions. It is believed that the dopamine transporter plays a key role in the mechanism of cocaine addiction. Because a good portion of our knowledge about drug addiction is derived from studying mouse as an animal model, it is essential to compare the properties of dopamine transporter from human and mouse. We report here the cloning of the mouse dopamine transporter (mDAT) cDNA and its expression and comparison with the human DAT. The 3.4 kilobase (kb) cDNA encodes a polypeptide that is 93.5% identical to the hDAT, with 619 amino acid residues and a calculated molecular weight of 68.8kDa. Dopamine transporters from mouse and human were stably expressed in the same parental MDCK cells and their properties were compared. The Michaelis-Menten constant Km values are 2.0 microM for mDAT and 2.4 microM for hDAT. Mouse and human DAT were also compared for drug inhibition profiles. Dopamine transporters from the two species have the same sensitivity to amphetamine (Kd: 0.75 microM) and bupropion (Kd: 1.5 microM). However, hDAT is more sensitive than mDAT to cocaine (Kd: 0.14 microM and 0. 29 microM respectively) and to ritalin (Kd: 0.038 microM and 0. 12 microM respectively). The cloning of mDAT cDNA provides an important tool for further study of the mechanism of drug addiction using mouse as an animal model. PMID- 10375633 TI - Cloning, mapping and tissue-specific expression of Drosophila clathrin-associated protein AP50 gene. AB - The Drosophila homologue of AP50, the medium chain of clathrin-associated protein complex AP-2, was identified and characterized from the Drosophila Expressed Sequence Tag database. The Drosophila AP50 is 86% identical to that of mouse and human, and 80% identical to the Caenorhabditis elegans homologue. It is a single copy gene with two mini-introns in the coding region and it maps to position 94B1 B2 on polytene chromosomes. Two P1 clones, DS01102 and DS0104, were identified that contain the AP50 gene. Alternative 5' UTR splicing is involved in the regulation of AP50 expression. AP50 expression is highly enriched in the central nervous system and midgut caecum during embryo development, and its function is discussed. The two other Drosophila members of the medium-chain family of clathrin-associated protein complexes, AP47 and mu3, have also been identified and mapped to 85D20-D27 and 6E1-E4, respectively. PMID- 10375634 TI - Analysis of expressed sequence tags of porcine skeletal muscle. AB - Porcine skeletal muscle genes play a major role in determining muscle growth and meat quality. Therefore, to progress towards a better understanding of the genetic factors influencing these traits, the first step is to characterize the genes expressed in skeletal muscle tissue in pig. To this aim, we constructed a porcine biceps femoris muscle cDNA library and sequenced 111 randomly isolated clones. By FASTA analysis, we identified 72 unique clones: 47 showed homology to previously identified genes in human or other mammals, 20 matched uncharacterized expressed sequence tags (ESTs), two showed no significant matches to sequences already present in DNA databases, and three other clones containing only repetitive elements were excluded from further analysis. Mitochondrial genes (16.2%), myosin heavy chain genes (9%) and the actin alpha skeletal muscle gene (9%) were the most abundant transcripts. Among the 47 identified genes several muscle-specific or predominant sequences expressed in skeletal muscle were found. The sequences of the clones matching uncharacterized human, mouse or porcine ESTs were tested by GRAIL in order to identify putative coding regions. The results of our analysis allowed the establishment of a first list of genes expressed in porcine skeletal muscle. PMID- 10375635 TI - Cloning, mapping and expression of UBL3, a novel ubiquitin-like gene. AB - A novel Drosophila melanogaster gene UBL3 was characterized and shown to be highly conserved in man and Caenorhabditis elegans (C. elegans). The human and mouse homologues were cloned and sequenced. UBL3 is a ubiquitin-like protein of unknown function with no conserved homologues in yeast. Mapping of the human and mouse UBL3 genes places them within a region of shared gene order between human and mouse chromosomes on human chromosome 13q12-13 and telomeric mouse chromosome 5 (MMU5). PMID- 10375636 TI - YAH1 of Saccharomyces cerevisiae: a new essential gene that codes for a protein homologous to human adrenodoxin. AB - Here we describe the identification of a yeast gene (YAH1) with significant homology to a mammalian enzyme, adrenodoxin, encoded in open reading frame (ORF) YPL252C. Adrenodoxin is the second electron carrier that participates in a mitochondrial electron transfer chain that, in mammals, catalyses the conversion of cholesterol into pregnenolone, the first step in the synthesis of all steroid hormones. The inactivation of the yeast gene's chromosomal copy reveals that it performs an essential function. We show that the protein is targeted to the mitochondrial matrix and describe attempts to complement the yeast knockout with the human adrenodoxin gene (FDX1) and with chimerical proteins constructed with the fusion of the yeast and the human gene. The previous identification of a homolog of the first mammalian enzyme in yeast, ARH1, also shown to be essential (Manzella, L., Barros, M.H., Nobrega, F.G., 1998. Yeast 14, 839-846), strongly suggests that there is a novel electron transfer chain, unlinked to respiration, and of essential function in mitochondria. PMID- 10375637 TI - A 450-kb contig of defensin genes on human chromosome 8p23. AB - Defensins are a large family of host defense peptides expressed in leukocytes and epithelia. Using P1 and BAC clones, we have determined the organization of the human alpha-defensin genes and the beta-defensin gene HDEFB1 on chromosome 8p23. From the telomere, the order of the genes (with encoded peptides in parentheses) is HDEFA5 (HD-5), HDEFA1/1A (HNP-1/3), HDEFA4 (HNP-4), HDEFA6 (HD-6), and HDEFB1 (HBD-1). These genes span a region of approximately 450kb. Genes encoding intestinal Paneth cell defensins (HDEFA5 and HDEFA6) flank the myeloid defensin gene cluster (HDEFA1, HDEFA1A, HDEFA4). Based on our previous studies, the remaining known defensin gene, HDEFB2 (HBD-2), is about 400kb centromeric to HDEFB1. This map supports the hypothesis, originally proposed because of sequence similarities, that myeloid alpha-defensin genes evolved by reduplication and divergence from Paneth cell defensin genes, and identifies regions and clones, which should be useful in the search for new defensin genes. PMID- 10375638 TI - Multiple 5'-untranslated exons in the nuclear respiratory factor 1 gene span 47 kb and contribute to transcript heterogeneity and translational efficiency. AB - Nuclear respiratory factor 1 (NRF-1) is a nuclear transcription factor that has been implicated in the nuclear control of respiratory chain expression in mammalian cells. Here, we demonstrate that a complex pattern of alternative splicing contributes to sequence heterogeneity within the human NRF-1 5' untranslated region (UTR). At least six different 5'-UTR exons (UTRs 1-6) were detected in NRF-1 transcripts. These exons were mapped to human NRF-1 genomic clones and their sequences, including donor and acceptor splice junctions, determined. Two of the human UTR exons were derived from insertions of Alu-sq family members into the NRF-1 locus. The distance between the transcription initiation sites in UTR1 and the first protein coding exon is approx. 47kb, bringing the total length of the human NRF-1 gene to approx. 104kb. In contrast to human, only two UTR exons were found in mouse. The mouse UTR1 sequence obtained is identical to human UTR1, but mouse UTR2 bears no resemblance to any of the human exons. Mutations within human UTR1 modulate NRF-1 expression by interfering with mRNA translational efficiency in transfected cells and in an in vitro translation system. The effects of the mutations are proportional to their ability to disrupt predicted mRNA secondary structures within UTR1. Thus, the unusually high sequence conservation within UTR1 in part reflects selective constraints on translational expression. PMID- 10375639 TI - Cloning and sequencing of an acetyl-CoA synthetase (ADP-forming) gene from the amitochondriate protist, Giardia lamblia. AB - A Giardia lamblia gene, Glacs, was cloned, sequenced and expressed in Escheria Coli. This gene codes for a 726 residue long acetyl-CoA synthetase (ADP-forming). This enzyme is responsible for the formation of acetate, a metabolic endproduct of G. lamblia. It is known from only two Type I amitochondriate eukaryotes, G. lamblia and Entamoeba histolytica and from the archaebacterium, Pyrococcus furiosus. With Glacs as query, homologous unidentified open reading frames were detected in the complete genomes of only a few archaebacteria and eubacteria. These form a new protein family present in all three domains of life, which probably plays a central role in the acyl-CoA metabolism but is of restricted taxonomic distribution. PMID- 10375640 TI - Identification, expression and chromosome localization of a human gene encoding a novel protein with similarity to the pilB family of transcriptional factors (pilin) and to bacterial peptide methionine sulfoxide reductases. AB - Here we report the isolation, characterization and chromosome localization of a subtracted cDNA (CBS-1) isolated from the human ocular ciliary body which encodes a novel protein. As is deduced from the nucleotide sequence of the cDNA, CBS-1 contains an open reading frame consisting of 182 amino acids, with a molecular weight of 19.5kDa. CBS-1 shares significant nucleotide and amino acid sequence identities (residues 51 to 182) with a hypothetical 15.5kDa protein in the ANSA GAP intergenic region (yeaA) of Escherichia coli, and the carboxyl terminal region of pilB, a transcription factor involved in the regulation of expression of pili, from Neisseria gonorrhoeae. Interestingly, CBS-1 also shares significant identity with the carboxyl terminus of the peptide-methionine sulfoxide reductase (MsrA), a repair enzyme, from Helicobacter pylori and Streptococcus pneumoniae. However, the amino terminal of CBS-1 (residues 23 to 43), which lacks homology to the amino terminal region of gonococcal pilB or pneumococcal MsrA, exhibits significant identity in a stretch of 20 amino acids, with glycine-rich proteins. By Northern blot, CBS-1, hybridized to a 0.6 to 0.7kb transcript in size, is expressed ubiquitously in many tissues, but most abundantly in the retina and ocular ciliary body, skeletal muscle and heart. An epitope-directed antibody to an amino acid sequence at the carboxyl terminus of CBS-1 recognized a main protein of 19.5kDa in ocular ciliary body extracts, and a 23kDa protein in total extracts from E. coli MC1061 cells, which expresses high levels of MsrA. The CBS 1 gene was mapped to human chromosome 10p12 between markers WI-8535 and WI-4724, and is tightly linked to the two STRP markers of D10S1789 and D10S550. We suggest that the CBS-1 gene encodes a mammalian transcription factor related to the bacterial pilB and certain bacterial MsrA homologues. PMID- 10375641 TI - Characterization of the Drosophila ortholog of mouse eIF-3p48/INT-6. AB - The mouse mammary tumor virus (MMTV) has been shown to integrate frequently into INT-6 in MMTV-induced mouse mammary tumors. The INT6 gene has been highly conserved through evolution and has recently been shown to encode the p48 component of the eucaryotic translation initiation factor 3 (eIF-3) complex. We report here the isolation of the Drosophila eIF-3p48/INT-6. The gene comprises three exons within 1.8kb of genomic DNA located at cytogenetic position 73C2 in the Drosophila genome. The 1.5kb eIF-3p48/INT-6 RNA species encodes a protein composed of 364 amino-acid residues whose sequence is 71% similar to that of the mouse/human eIF-3/INT-6 amino-acid sequence. eIF-3p48/INT-6 RNA is expressed throughout development in Drosophila and the encoded protein is associated with the microsomal subcellular fraction. PMID- 10375642 TI - Interspersed DNA element restricted to a specific group of telomeres in the dipteran Chironomus pallidivittatus. AB - Telomeres in the dipteran Chironomus pallidivittatus terminate with 340bp tandem DNA repeats belonging to different subfamilies with characteristic intertelomeric distribution. We have now found, interspersed between such repeats, a composite element of approx. 1400bp present in two similar size variants, with several components of nontelomeric origin. There were about 50 copies of the element, predominantly or exclusively present in a previously defined group of telomeres, characterized by a unique set of telomeric tandem repeat subfamilies. Elements were integrated at irregular distances from each other, and intervening telomeric tandem repeat DNA was variable in composition. Nevertheless, the flanks immediately surrounding the elements were identical for different elements; in other words, there was a site-specific insertion. We suggest that this selective invasion of a small part of the genome by an interspersed, probably rapidly evolving element is best explained by repeated gene conversions. PMID- 10375643 TI - Cloning and characterization of a bifunctional RelA/SpoT homologue from Mycobacterium tuberculosis. AB - A 2.2kb relA/spoT homologue was isolated from Mycobacterium tuberculosis (Mtb) genomic DNA by PCR-amplification. The Mtb gene encodes a protein of 738 amino acid residues, and is flanked upstream by an ORF that is highly similar to the apt gene, and downstream by an ORF that is highly similar to the cypH gene. This dual function Mtb homologue belongs to the relA/spoT family of genes that mediate the stringent response by regulating the synthesis and degradation of guanosine 3',5'-bis(diphosphate) (ppGpp) and pppGpp. In vitro biochemical data indicate that purified RelMtb is a ribosome- and tRNA-independent ATP:GTP/GDP/ITP 3' pyrophosphoryltransferase. Additionally, purified RelMtb is an Mn2+-dependent, ribosome and tRNA-independent, (p)ppGpp 3'-pyrophosphohydrolase. These reactions were also assessed in vivo in E. coli deleted in both the relA and spoT genes, which generates a (p)ppGpp0 phenotype. RelMtb can suppress this phenotype and can generate more (p)ppGpp than relA in the wild type E. coli control. PMID- 10375644 TI - Sequencing and exon mapping of the inositol 1,4,5-trisphosphate receptor cDNA from Drosophila embryos suggests the presence of differentially regulated forms of RNA and protein. AB - A single gene appears to code for the inositol 1,4,5-trisphosphate receptor (itpr) in Drosophila melanogaster, as compared to three known genes in mammals. Expression of the itpr gene in Drosophila occurs in a wide range of tissues and developmental stages, suggesting its requirement during diverse cellular and physiological processes. A head cDNA for the Drosophila IP3R has previously been cloned and sequenced. Here we present and analyse the sequence of cDNAs encoding the complete IP3R, obtained from embryonic stages. The embryonic cDNA is 10525bp long and is a splice variant of the head cDNA. It differs from the latter in three main respects. It has longer 5' and 3' untranslated regions, two potential casein kinase II sites are missing in the embryo form and it contains an alternate exon which results in the replacement of three residues (VHF) in the head form by five residues (GVGHSV) in the embryo form. The significance of these changes is discussed. An exon-intron map of the gene derived from sequencing of intron-containing genomic fragments is also presented. The gene has a total of 11 introns, of which more than half are clustered in a region of the modulatory domain of the IP3R. PMID- 10375645 TI - Galanin antagonizes vasopressin-stimulated flank marking in male golden hamsters. AB - Microinjection of vasopressin (VP) into the anterior hypothalamus (AH) of golden hamsters induces a rapid bout of flank marking, a stereotyped scent marking behavior used for olfactory communication. In rats, VP is colocalized with galanin (GAL) in several brain regions. GAL has been shown to antagonize the postsynaptic actions of other cosecreted neurotransmitters including acetylcholine and norepinephrine; however, the ability of GAL to modulate the postsynaptic actions of VP has not been assessed. Here, we report that coadministration of GAL can block VP-induced flank marking in golden hamsters in a dose dependent manner. These findings provide the first evidence in any species that GAL can antagonize the central actions of VP. Using slice binding and receptor autoradiography, we have identified GAL binding sites in the AH and two other regions implicated in flank marking behavior (the lateral septum and central grey). These findings raise the possibility that endogenous GAL may function as an inhibitory modulator of this stereotypic scent marking behavior. PMID- 10375646 TI - Kynurenate attenuates the accumulation of diacylglycerol and free fatty acids after experimental brain injury in the rat. AB - This study examined the effects of the administration of kynurenate, a non specific excitatory amino acid (EAA) receptor subtype antagonist, on the regional accumulation of diacylglycerol (DG) and free fatty acids (FFAs) after lateral fluid percussion (FP) brain injury in the rat. After brain injury of moderate severity (2.0 atm), rats were treated with either kynurenate (200 mg/kg, i.v.) or saline at 5 min after injury. In the saline-treated brain-injured rats, levels of all individual DG-fatty acids (palmitic, stearic, oleic and arachidonic acids) and total DG-fatty acids were increased in the ipsilateral left cortex and hippocampus at 30 min and 60 min after injury. Kynurenate administration attenuated increases of individual and total DG-fatty acids in the ipsilateral cortex at 30 and 60 min and in the ipsilateral hippocampus at 30 min after FP brain injury. At 30 and 60 min after FP brain injury, increases in the levels of individual FFAs (palmitic, stearic, oleic and arachidonic acids) and of total FFAs were also observed in the ipsilateral cortex and hippocampus of the saline treated injured rats. Kynurenate administration attenuated increases of all individual and total FFAs in the ipsilateral cortex and hippocampus either at 30 min alone or at both 30 min and 60 min after FP brain injury. In the contralateral cortex, levels of both DG-fatty acids and FFAs were not increased in the saline-treated injured rats and were also not affected by the administration of kynurenate. These results support the role of EAA receptor subtypes in the phospholipases-catalyzed formation of DG and FFAs in the ipsilateral cortex and hippocampus after lateral FP brain injury. PMID- 10375647 TI - Neural mechanisms of soleus H-reflex depression accompanying voluntary arm movement in standing humans. AB - This study have investigated the changes in soleus (Sol) H-reflexes by arm movement during freely standing (FS) and back-supported standing (BS) in healthy subjects. Before the arm movement, there is an anticipatory phase, which includes increased electromyographic (EMG) activity in the biceps femoris (BF) and decreased EMG activity of the Sol muscle. The Sol H-reflex appeared to be inhibited during the anticipatory phase as well as during the time of arm movement. However, the inhibition appeared to be larger in FS than in BS conditions. Vibration applied to the tendon of the BF muscle depressed the Sol H reflex. This inhibition was attributed to presynaptic inhibition and was reduced during the anticipatory phase, and was not very much changed during arm movements. It is suggested that the depression of the Sol H-reflex induced by voluntary arm movement has two inhibitory components of different origins. Descending motor commands generate the early inhibitory component, while the late component is produced by the presynaptic inhibition that results from peripheral inputs. The inhibition related to anticipatory postural adjustment (APA) indicates that a new-setting of the spinal mechanisms is required and responsible in order to stabilize body equilibrium which is dependent upon different postural conditions. PMID- 10375648 TI - Excitatory effect of Cd2+ on cat adrenal chromaffin cells. AB - To understand the mechanism(s) underlying the Cd2+- and Co2+-induced increases in the cytosolic free Ca2+ concentration ([Ca]i) in cat adrenal chromaffin cells, we used nystatin-perforated patch recording method and fura-2 microfluorometry. Under the current-clamp conditions, the external application of 5x10(-7) M Cd2+ slowly depolarized the cells resulting in the bursting of action potentials. Under the voltage-clamp conditions, Cd2+ evoked a slow inward current accompanied by a decrease of K+ conductance at a holding potential of -40 mV, and Co2+ mimicked Cd2+ action. In some cells (16%), Cd2+ evoked an additional rapid transient outward current associated with an increased K+ conductance and a successive slow inward current. The Cd2+-induced inward current was activated in a concentration-dependent manner with a half-maximum concentration of 9.3x10(-8) M. The Cd2+- and Co2+-induced [Ca]i increases measured with fura-2 microfluorometry were maximal at 10(-6) and 10(-5) M, respectively, and the higher concentrations of both cations caused the smaller responses. Additional transient increase in [Ca]i was often evoked upon the removal of relatively higher concentrations of these metals. It was concluded that the Cd2+-induced membrane depolarization due to the decrease in K+ conductances evoked the bursting firings resulting in the increase in [Ca]i, and consequently might stimulate the catecholamine secretion. PMID- 10375649 TI - Reversible attenuation of glutamatergic transmission in hippocampal CA1 neurons of rat brain slices following transient cerebral ischemia. AB - The present experiments were conducted to determine the time course of synaptic dysfunction in the vulnerable regions of the post-ischemia hippocampus. Following transient cerebral ischemia, neurons in the CA1 subfield of the hippocampus undergo a delayed degeneration that develops about 48 h after reperfusion. We have shown previously that CA1 glutamatergic transmission is decreased in the CA1 subfield well before any morphological deterioration of the CA1 cells is visible under the light microscope. However, it is unknown whether a time window exists after insult in which attenuated synaptic activity may be restored to normal levels. We show here that evoked CA1 somatic population spikes and dendritic field potential responses decline progressively after reperfusion in the CA1 subfield, such that by 72 h post-insult, the challenged neurons are unable to elicit evoked excitatory responses. This attenuation of synaptic transmission was confined to the vulnerable neurons of the hippocampus, however, as the evoked responses in the dentate gyrus displayed amplitudes that were not significantly diminished from sham control after challenge. In brain slices obtained from 24 h post-ischemia rats with significantly impaired CA1 somatic responses, the application of 5 or 50 microM of the potassium channel blocker 4-aminopyridine (4 AP) restored the magnitude of the evoked excitatory response to control values. At 36 h post-ischemia, the decreased CA1 evoked responses could be partially improved by 4-AP, but not to control levels. Based upon these results, we conclude that the decreased CA1 synaptic activity at 24 h post-ischemia is potentially reversible, and suggest that 4-AP improves the CA1 synaptic responses at least in part by improving transmitter release at post-ischemia glutamatergic synapses. PMID- 10375650 TI - Biondi ring tangles in the choroid plexus of Alzheimer's disease and normal aging brains: a quantitative study. AB - The choroid plexus (CP) performs the vital function of producing up to 90% (450 1000 ml/day) of cerebrospinal fluid (CSF) to nourish and to protect the brain in the CSF suspension. The CP also acts as a selective barrier between blood and CSF to regulate ions and other essential molecules. However, the accumulation of intracellular inclusions called Biondi ring tangles (BRTs) in CP cells of Alzheimer's disease (AD)/aging brains may affect these vital functions of the CP. Statistical analysis of quantitative data on the numbers of CP cells containing BRTs from 54 brains (29 AD and 25 normal control), age range 1-100 years, indicated a significant difference (p<0.00004) between AD and control brains, using analysis of covariance (ANCOVA) with age as covariate. This study compiled the first set of archives to reveal the distribution pattern of BRTs in the CP of AD brains at various ages. Electron microscopy of negatively stained isolated BRTs revealed that these tangles are made of tightly packed bundles of long filaments with diameter around 10 nm that are morphologically distinct from the more loosely packed/shorter bundles of 6-8 nm amyloid fibrils of neuritic plaques (NPs) and from the 24 nm paired helical filaments of neurofibrillary tangles (NFTs) in AD brain. These data suggest that BRTs may represent a significant and measurable biomarker for AD in addition to NPs and NFTs. PMID- 10375651 TI - Colocalization of VIP with AVP in neurons of the human paraventricular, supraoptic and suprachiasmatic nucleus. AB - Aim of this study was to investigate, with the aid of a recently developed immunofluorescence technique, cellular colocalization of vasoactive intestinal peptide (VIP) with arginine-vasopressin (AVP) in the paraventricular nucleus (PVN), the supraoptic nucleus (SON) and the suprachiasmatic nucleus (SCN) of the human hypothalamus. To this end, six hypothalami resected from patients who had died suddenly served as material of research. After formaldehyde fixation and subsequent storage in 30% sucrose, 25-microm thick cryosections were cut of one half of each hypothalamus. These sections were double-immunolabeled with primary antibodies against AVP and VIP followed by fluorophore-conjugated secondary antibodies. Autofluorescence, mainly caused by lipofuscin granules in neurons and glial cells, was blocked by a specially developed procedure consisting of incubating the immunolabeled sections in a Sudan Black B solution. Quantitative analysis with a confocal laser scanning microscope showed that of all stained cellular profiles the percentages of profiles immunoreactive exclusively for AVP or VIP or for both neuropeptides (colocalization) were for the SCN approximately 76.5%, 19.6% and 3.9%, for the SON 97.7%, 0.2% and 2. 1% and for the PVN 93.2%, 1.6% and 5.2%, respectively. These data illustrate that colocalization between AVP and VIP is not only present in neurons of the PVN and SON, but also in neurons of the SCN. This unexpected finding illustrates that the human SCN may also use a highly differentiated language to transmit its circadian signal to the rest of the brain. PMID- 10375652 TI - Microglia activation in a model of sleep disorder: an immunohistochemical study in the rat brain during Trypanosoma brucei infection. AB - Microglial cells play a key role in the events triggered by infection, injury or degeneration in the central nervous system not only as scavenger cells but also as immune effector elements. We analyzed the features and distribution of cells of the microglia/macrophage lineage with OX-42 and ED-1 immunohistochemistry in the brain of experimental rats infected with the extracellular parasite Trypanosoma brucei. Such experimental infection provides a rat model of sleeping sickness or African trypanosomiasis, and is hallmarked in its advanced stages by severe alterations of the animals' sleep structure. In infected rats a remarkable activation of microglia, revealed by OX-42 immunoreactivity, became evident in the 3rd week post-infection in periventricular and subpial brain regions, with a prevalence in the hypothalamus. These features were concomitant with the onset of sleep anomalies, monitored with electroencephalographic recordings. Microglia activation increased in the following weeks, paralleling the progressive alterations of sleep parameters, and was most marked in the terminal stages of the infection, corresponding to the 6th-7th weeks. In addition, ED-1 immunoreactive macrophages and ramified microglia, confined to hypothalamic periventricular and basal regions, were evident after 4 weeks of disease. Degeneration of neuronal perikarya was not detected histologically in the infected brains at any time point. These data provide evidence for a reaction of microglia and macrophages in the brain of trypanosome-infected rats, and point out a selective distribution of these activated cells. The findings are discussed in relation to the animals' sleep disorder during trypanosome infection. PMID- 10375653 TI - Neuropeptides, nitric oxide synthase and GAP-43 in B4-binding and RT97 immunoreactive primary sensory neurons: normal distribution pattern and changes after peripheral nerve transection and aging. AB - We have here sought to cross-correlate the expression of immunoreactivities for several neuropeptides, nitric oxide synthase (NOS) and the growth associated protein GAP-43 in subpopulations of dorsal root ganglion (DRG) neurons tagged by the selective markers isolectin B4 and the neurofilament antibody RT97, selective for, respectively, subpopulations of small and large DRG neurons. By use of double- and triple-labeling immunohistochemistry, non-manipulated and sciatic nerve transected young adult rats as well as aged (30-months-old) rats were examined using a confocal microscope equipped with enhanced spectral separation. In young adult rats, the DRG neuron profiles could be divided into three subpopulations (B4 binding (B4+) approximately 50%; RT97-immunoreactive (RT97+) approximately 35%; B4-/RT97- approximately 15%). Calcitonin gene-related peptide (CGRP) is expressed in all three subpopulations. Galanin message-associated peptide (GMAP) colocalize with CGRP (100%) but is not expressed in RT97+ profiles. NOS is present in the RT97- subpopulations and frequently colocalize with CGRP (92%). GAP-43 is expressed in all three DRG subpopulations and colocalize with CGRP (88%), GMAP (38%) and/or NOS (22%). Only very small differences were seen among the young adult rats, implicating that the size of respective subpopulation as well as the expression pattern for neuropeptides, NOS and GAP-43 are fairly stable. Sciatic nerve transection reduced B4-binding but not RT97-like immunoreactivity. Distinct changes in the expression of neuropeptides, NOS and GAP-43 were evident in the DRG subpopulations and, furthermore, the regulatory changes were very similar among the lesioned animals. The relative size of the DRG subpopulations was unaffected by aging, while the expression of neuropeptides was altered showing similarities with the changes induced by axotomy in young adult rats. PMID- 10375654 TI - Peripheral injections of Freund's adjuvant in mice provoke leakage of serum proteins through the blood-brain barrier without inducing reactive gliosis. AB - Breakdown of the blood-brain barrier (BBB) and ensuing gliosis are common events following physical trauma to the central nervous system (CNS) or during autoimmune diseases such as experimental allergic encephalomyelitis (EAE). Some studies of EAE in rodents report that peripheral injections of complete Freund's adjuvant (CFA), which contains heat-inactivated Mycobacterium to provoke peripheral inflammation without adversely affecting the CNS, can itself lead to increased BBB permeability to small tracer molecules and certain serum proteins. To study the equivocal relationship between serum protein extravasation and reactive gliosis, we injected C57BL/6 mice with CFA and histologically assessed the permeability of various serum proteins and the reactivity of proximal microglia and astrocytes in the uninjured brainstem and spinal cord enlargements after 1-4 weeks. Our results confirm that CFA injections induce progressive increases in the perivascular extravasation of serum IgG, albumin, IgM, and exogenous horseradish peroxidase, all to varying degrees, most prominently in the brainstem and cervical spinal cord after 2-3 weeks. More importantly, neither microglial cells nor astrocytes in regions of focal serum protein leakage appeared morphologically reactive based on immunoreactivity for CR3 receptors (Mac-1) or glial fibrillary acidic protein (GFAP), respectively. Because we found no evidence of T cell infiltration accompanying the exudates, our results indicate that in the absence of physical trauma or inflammatory cells resident CNS neuroglia remain quiescent upon exposure to extravasated serum proteins. PMID- 10375655 TI - Stimulation of the chewing area of the cerebral cortex induces inhibitory effects upon swallowing in sheep. AB - Mastication and swallowing are two tightly integrated components of food intake behavior. We investigated the effects of stimulating the chewing area of the fronto-orbital cortex (CCx) on some muscles and medullary interneurons (Ins) or motoneurons (Mns) active during swallowing. For the purpose of comparison, the lingual nerve (LN) was also stimulated during the experiments. Electromyography (EMG) and extracellular neuronal recording were used, and swallowing was reflexly induced (RIS) by stimulation of the superior laryngeal nerve (SLN). RIS was almost totally abolished during long-lasting repetitive stimulation of CCx or LN, and was strongly facilitated after stimulation cessation. Short-duration stimulation (one or a few pulses) of both the CCx and LN also inhibited triggering of deglutition when delivered just before the onset of RIS. This inhibition appeared as a delay or total suppression of the EMG and neuronal swallowing activities. It was obvious at the level of the muscles, the hypoglossal Mns and the premotoneurons (PMns; Ins of the ventral medulla near the nucleus ambiguus), as well as at the level of the Ins of the dorsal medulla (within or around the solitary tract nucleus) which are assumed to be the core of the 'central pattern generator' (CPG) for swallowing. In addition to the 'chewing related inhibition', many ventral Ins exhibited a short latency synaptic activation after CCx and/or LN stimulation. Therefore, these Ins may play a pivotal role for reflex or cortical fast control of tongue (and jaw) muscles, and for coordinating their contractions in the context of mastication-deglutition interactions. PMID- 10375656 TI - Suppression of post-ischemic-induced fos protein expression by an antisense oligonucleotide to c-fos mRNA leads to increased tissue damage. AB - Activation of c-fos, an immediate early gene, and the subsequent upregulation of Fos protein expression occur following neural injury, including focal cerebral ischemia (fci). Fos and Jun form a heterodimer known as activator protein 1, which regulates the expression of many late effector genes. To study the downstream effects of c-fos expression following ischemia, we suppressed the translation of c-fos by administering an antisense oligonucleotide (AO) to c-fos mRNA. Eighteen hours prior to fci, male, Long Evans (LE) rats received intraventricular injections of AO, mismatched AO (MS) or artificial cerebrospinal fluid (aCSF). Fci was induced by permanent right middle cerebral artery occlusion. At 24-h post-occlusion, neurological function was assessed, and the animals were sacrificed. The brains were removed and stained with triphenyltetrazolium chloride for infarct volume determination. Fos immunohistochemistry was performed in separate animals to determine the effects of treatment on Fos expression number of Fos positive cells. AO administration reduced the number of cells with fci-induced Fos expression by approximately 75%. No differences in neurological scores existed between any of the groups. AO treated LE developed larger infarcts (40.1+/-1.0%, mean+/-S.D., p<0.001) than MS- or aCSF-treated controls (34.3+/-1.0%, 34.6+/-1.0%, respectively). These results suggest that c-fos activation and subsequent Fos protein expression exerts a neuroprotective effect, which is likely via upregulation of neurotrophins, following focal cerebral ischemia. This response, among others, may contribute to brain adaptation to injury that underlies functional recovery after stroke. PMID- 10375657 TI - FK960 [N-(4-acetyl-1-piperazinyl)-p-fluorobenzamide monohydrate], a novel potential antidementia drug, restores the regional cerebral blood flow response abolished by scopolamine but not by HA-966: a positron emission tomography study with unanesthetized rhesus monkeys. AB - The interactions of FK960 [N-(4-acetyl-1-piperazinyl)-p-fluorobenzamide monohydrate], a novel potential antidementia drug, with cholinergic and glutamatergic neuronal systems were evaluated with respect to its effects on the regional cerebral blood flow (rCBF) response to vibrotactile stimulation in unanesthetized rhesus monkeys with [15O]H2O and high resolution positron emission tomography (PET). Under a saline condition, the vibrotactile stimulation given on the right forepaw induced a significant increase in the rCBF response in the contralateral somatosensory cortex of the monkey brain. Systemic administration of scopolamine (50 microg/kg, i.v.), a muscarinic cholinergic receptor antagonist, completely abolished the rCBF response to the stimulation, and the abolishment lasted, at least, up to 4 h after scopolamine injection. The scopolamine-induced abolishment of rCBF response was restored by the administration of FK960 at relatively wide dosing range from 1 to 1000 microg/kg (i.v. ), and the recovery by FK960 on the rCBF response lasted for 1 h following the administration of FK960 at doses of 100 and 1000 microg/kg. On the other hand, the rCBF response abolished by 1000 microg/kg of (+)-3-amino-1-hydroxy-2 pyrrolidone (HA-966), an antagonist of the glycine modulatory site on the N methyl-d-aspartate (NMDA) receptors, was not restored by FK960 (1000 microg/kg, i.v.). These findings suggest that FK960 reverses the abolished rCBF response to somatosensory stimulation via enhancement of cholinergic neurotransmission but not via the glutamatergic one. PMID- 10375658 TI - Postnatal ethanol exposure blunts upregulation of GABAA receptor currents in Purkinje neurons. AB - Recently, we found that early postnatal ethanol exposure inhibits the maturation of GABAA receptors (GABAARs) in developing medial septum/diagonal band (MS/DB) neurons, suggesting that these receptors may represent a target for ethanol related to fetal alcohol syndrome (FAS). To determine whether GABAARs on other neurons are also sensitive to a postnatal ethanol insult, postnatal day (PD) 4-9, rat pups were artificially reared and exposed to ethanol (4.5 g kg-1 day-1, 10.2% v/v). The pharmacological profile of acutely dissociated cerebellar Purkinje cell GABAARs from untreated, artificially reared controls and ethanol-treated animals was examined with conventional whole-cell patch clamp recordings during PD 12-16 (juveniles) and PD 25-35 (young adults). For untreated animals, GABA (0.3-100 microM) consistently induced inward Cl- currents in a concentration-dependent manner showing an age-related increase in maximum response without change in EC50 or slope value. Acute ethanol (100 mM) consistently inhibited 3 microM GABA currents (10-20%); positive modulators, pentobarbital (10 microM), midazolam (1 microM) and loreclezole (10 microM), consistently potentiated; the negative modulator, Zn2+ (30 microM), inhibited GABA currents across both juvenile and young adult groups. Loreclezole potentiation increased while Zn2+ inhibition decreased with age in untreated Purkinje neurons. Postnatal ethanol exposure (PD 4-9) decreased GABAAR maximum current density in young adult Purkinje cells but not in juvenile neurons. However, sensitivity to allosteric modulators did not change after ethanol. These data are consistent with the hypothesis that postnatal ethanol exposure during the brain growth spurt can disturb GABAAR development across the brain, although the mechanism(s) underlying this action remains to be determined. PMID- 10375659 TI - Generation of aberrant sprouting in the adult rat brain by GAP-43 somatic gene transfer. AB - The expression of GAP-43 was modulated genetically in the adult rat nigrostriatal or septohippocampal pathway using recombinant adeno-associated virus (rAAV) vectors incorporating the neuron specific enolase (NSE) promoter and either a rat GAP-43 cDNA or the corresponding antisense sequence. Bicistronic expression of green fluorescent protein (GFP) enabled us to evaluate transduced neurons selectively. Single injections of rAAV into the substantia nigra pars compacta (SNc) transduced both dopaminergic and non-dopaminergic neurons stably for the 3 month duration of the study. Transduction with the GAP-43 vector in this region: (1) increased GAP-43 mRNA levels 2-fold compared to controls; (2) led to GAP-43 immunoreactivity in neuronal perikarya, axons, and dendrites that was not observed otherwise; and (3) resulted in GAP-43/ GFP-positive axons that were traced to the striatum where they formed clusters of aberrant nets. The GAP-43 antisense vector, in contrast, decreased neuropil GAP-43 immunoreactivity compared to controls in the SNc. In septum, injections of the GAP-43 expressing vector also caused aberrant clusters of GAP-43 labelled fibers in terminal fields, i.e., fornix and hippocampus, that were not observed in control tissues. It therefore appears that rAAV vectors provide a novel approach for modulating intraneuronal GAP-43 expression in the adult brain. PMID- 10375660 TI - GABAA receptor stimulation blocks NMDA-induced bursting of dopaminergic neurons in vitro by decreasing input resistance. AB - The effects of the GABAA agonist, isoguvacine, on NMDA-induced burst firing of substantia nigra dopaminergic neurons were studied with intracellular and whole cell recordings in vitro. NMDA application caused the neurons to fire in rhythmic bursts. Although the NMDA-induced bursty firing pattern was insensitive to hyperpolarization by current injection, it was reversibly abolished by the selective GABAA agonist, isoguvacine. The block of the rhythmic burst pattern by isoguvacine application occurred regardless of whether the chloride reversal potential was hyperpolarizing (ECl-=-70 mV) or depolarizing (ECl-=-40 mV). In either case, the input resistance of the dopaminergic neurons was dramatically decreased by application of isoguvacine. It is concluded that GABAA receptor activation by isoguvacine disrupts NMDA receptor-mediated burst firing by increasing the input conductance and thereby shunting the effects of NMDA acting at a distally located generator of rhythmic burst firing. PMID- 10375661 TI - Strain differences in mesotelencephalic dopaminergic neuronal regulation between Fischer 344 and Lewis rats. AB - Differences in the behavioral responses of Lewis and Fischer (F344) inbred rat strains to stress and psychoactive drugs have been related to differences in the expression of various regulatory proteins in regions containing mesolimbic dopamine (DA) neurons. The present study compared basal and stimulated neurochemical estimates of DA utilization and synthesis in mesocortical, mesolimbic and nigrostriatal DA terminal regions of these two strains. In unstressed control animals, the Lewis strain had lower DA concentrations in the dorsal striatum (ST; 80.3% of F344) and lower basal dihydroxyphenylalanine (DOPA) accumulation after m-hydroxybenzylhydrazine (NSD 1015) treatment in the medial prefrontal cortex (mPfx; 75.3% of F344). Similar differences were observed in vehicle-injected animals. No strain differences in basal neurochemistry were apparent in the nucleus accumbens shell (NAs) or core (NAc). In response to restraint stress, dihydroxyphenylacetic acid (DOPAC) to DA ratios in the mPfx, NAs and ST increased in the F344 but not the Lewis strain. However, restraint stress did not significantly increase DOPA accumulation in the F344 strain. This latter finding was not due to a deficit in synthesis capacity, as gamma hydroxybutyric acid lactone (GBL) increased DOPA accumulation significantly more in F344 than Lewis animals. Finally, haloperidol increased DA utilization similarly in the two strains. Together these findings suggest that the inbred, behaviorally divergent F344 and Lewis rats have selective differences in mesocortical, nigrostriatal and mesolimbic DA neuronal regulation. PMID- 10375662 TI - Oxidative damage and breakage of DNA in rat brain after transient MCA occlusion. AB - As thrombolytic therapy for treatment of ischemic stroke was propagated, much attention has been paid to reperfusion brain injury. Oxidative stress is one of the most important factors that exacerbate tissue damage by reperfusion. Thus, we investigated the extent of oxidative damage in rat brain after transient middle cerebral artery (MCA) occlusion by immunohistochemical analysis for 8-hydroxy-2' deoxyguanosine (8-OHdG), which is one of the best markers of oxidative damage. Furthermore, in order to investigate its role in neuronal cell death, we performed terminal deoxynucleotidyl transferase-mediated dUTP-biotin in situ nick end labeling (TUNEL) study, and compared the results with that of 8-OHdG immunohistochemistry. There was no immunoreactive 8-OHdG in sham-operated brain, but it became present in neurons of MCA territory at 3 h of reperfusion after 90 min ischemia. At 48 h after reperfusion, cerebral tissue of MCA territory was severely destroyed, and many cells in that area revealed TUNEL positivity. Some neurons in MCA territory showed mild immunoreactivity for 8-OHdG at that time, but it was strongest in neurons in the outer area of MCA territory. Those cells did not show TUNEL positivity, suggesting that 8-OHdG production is not necessarily followed by early cell death. Here, it was demonstrated that oxidative DNA damage occurs in more extended area than that where cell death is recognized. Although this damage does not cause early cell death, this might result in more prolonged cell dysfunction and eventual neuronal loss. Anti oxidant therapy might be required for treatment of stroke in the future. PMID- 10375663 TI - An investigation on the role of nitric oxide in the modulation of the binding characteristics of various radioligands of GABAA receptors by 5alpha-pregnan 3alpha-ol-20-one in the rat brain regions. AB - Chronic administration of Nomega-nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide synthase, diminished the ability of 5alpha-pregnan 3alpha-ol-20-one, a neurosteroid, to potentiate the [3H]muscimol (5 nM) binding in the rat hippocampus but not in the cerebellum or cerebral cortex. This effect of NAME was stereospecific and susceptible to reversal by the pre-treatment of rats with L-arginine. However, chronic administration of L-NAME did not affect the modulation of the [3H]flunitrazepam (2 nM) or [35S]TBPS (4 nM) binding by the neurosteroid in any of the brain regions investigated in this study. These results suggest that nitric oxide may be involved in some of the effects of neurosteroids in hippocampus. PMID- 10375664 TI - Orphanin FQ/nociceptin blocks acquisition of morphine place preference. AB - Orphanin FQ/nociceptin (OFQ/N) suppresses the activity of the dopaminergic mesolimbic reward pathway yet reportedly fails to produce conditioned place aversion or preference. The present study sought to determine if this peptide could attenuate the development of morphine place preference. Male rats were administered OFQ/N (3 to 30 nmol intracerebroventricularly) during the induction of morphine (3 mg/kg subcutaneously) place preference. Animals receiving 3 or 10 nmol (but not 30 nmol) OFQ/N showed significant reductions in the development of place preference to morphine. PMID- 10375665 TI - Evidence for the collateral innervation of the esophagus and the heart from neurons in the compact formation of the nucleus ambiguus of the rat. AB - We investigated whether the heart receives collateral projections from the neurons which innervate the esophagus with a retrograde double-labeling method using two fluorescent tracers. Following injections of True Blue (TB) into the esophagus and Diamidino Yellow (DY) into the heart, about 21.9% of the labeled esophageal motoneurons in the compact formation of the nucleus ambiguus (AmC) were retrogradely double labeled. No single-labeled cardiac motoneurons were found in the AmC. The present results provide anatomical substrates for the esophagocardiac reflex. PMID- 10375666 TI - Postnatal development of action potential-induced dendritic calcium entry in neocortical layer II/III pyramidal cells. AB - By whole-cell patch-clamp recording and calcium imaging with fura-2, we investigated postnatal development of intracellular calcium dynamics in apical dendrites of layer II/III pyramidal cells in the rat visual cortex. Dendritic calcium increases, induced by single action potentials, occurred only slightly on postnatal days 8-10 (P8-10), and then underwent a gradual enhancement during the second postnatal week to become 2-3-fold larger on P16-18 than on P8-10. The results suggest that the developmental growth of calcium dynamics may play a critical role for functional development of neocortical neurons. PMID- 10375667 TI - Enhancement of activity-dependent calcium increase by neurotrophin-4 in visual cortex pyramidal neurons. AB - In pyramidal neurons from rat visual cortex slices, bath-application of NT-4 (20 ng/ml) did not much affected the baseline calcium signal, but did enhance calcium signals elicited by injections of depolarizing currents (+0.5 nA, 1 s). This enhancing effect of NT-4 was abolished by co-applying K252a. With ryanodine injected intracellularly, the effect of NT-4 was significantly reduced, suggesting an involvement of intracellular calcium release in this NT-4-induced enhancement of calcium transient. PMID- 10375668 TI - Dopamine transporter loss and clinical changes in MPTP-lesioned primates. AB - Single photon emission computed tomography (SPECT) and the dopamine (DA) transporter tracer, 2 beta-carboxymethoxy-3 beta-(4-iodophenyl)tropane ([123I]beta-CIT), were used to determine DA transporter density in 1-methyl-4 phenyl-1,2,3, 6-tetrahydropyridine (MPTP)-lesioned monkeys with varying degrees of parkinsonism. The clinical stage of parkinsonism corresponded to SPECT measures of striatal DA transporter density suggesting that more severe parkinsonism was associated with a greater degree of dopaminergic terminal degeneration. These findings are similar to those reported earlier using positron emission tomography (PET) and the DA metabolism tracer, 6-[18F]fluoro-L-m tyrosine (FMT), indicating that both are good methods for evaluating nigrostriatal degeneration in MPTP primate models. PMID- 10375669 TI - Bcl-2 transgenic mice with increased number of neurons have a greater learning capacity. AB - Transgenic mice overexpressing Bcl-2 in their neurons have an increased number of neurons. To assess whether this increased number of neurons leads to increased learning capacity we have used the Hebb-Williams maze which provides a measure of learning suitable for the study of small animals. We have demonstrated that bcl-2 transgenic mice learn faster and are more accurate in this maze. They required fewer trials to complete the maze and committed fewer errors. The transgenic mice were also faster than the wildtype mice, in particular the older mice. Prior to learning both groups of mice behaved in a similar way. These results show that bcl-2 transgene expression enhances learning capacity in mice by increasing the number of neurons. PMID- 10375670 TI - Effects of adrenergic stimulus on the activities of Ca2+ and K+ channels of dorsal root ganglion neurons in a neuropathic pain model. AB - We hypothesized that abnormal activity and adrenergic sensitivity in injured dorsal root ganglion (DRG) neurons are due to an intrinsic alteration of the cell body membrane. We investigated the effects of adrenergic stimulus on the activities of Ca2+ and K+ channels of DRG neurons in a rat chronic constriction injury (CCI) model. At first, we demonstrated thermal hyperalgesia and sprouting sympathetic nerve fibers in the ipsilateral L4-L5 DRGs. Using whole-cell patch clamp techniques, we found that alpha2-adrenergic stimulus by 10 microM norepinephrine (NE) inhibited inward currents (IBa, Ba2+ as a charge carrier) through voltage-dependent Ca2+ channels (VDCCs) of DRGs in the CCI model by 42%, whereas it enhanced the IBa by 18% in control animals. The inhibitory effect of NE disappeared by pretreatment with the N-type VDCC antagonist omega-conotoxin GVIA (1 microM). NE shifted the inactivation curve to a more negative potential, showing that it has inhibitory effects on IBa both in activated and in inactivated states. alpha2-Adrenergic stimulus also inhibited outward K+ currents by 24% in the CCI model, while it had no effect on the currents in control animals. The inhibitory effect of NE was blocked by pretreatment with the Ca2+ activated K+ (KCa) channel antagonist charybdotoxin (40 nM). The NE-induced inhibitory effects both on N-type VDCC and on KCa channels in injured DRG neurons of the CCI model could lead to cell membrane depolarization, resulting in a spontaneous discharge of action potential and an increase in sensitivity to adrenergic stimulus. PMID- 10375671 TI - Characteristics of the muscle spindle endings of the masticatory muscles in the rabbit under halothane anesthesia. AB - To explore the response characteristics of muscle spindle units in the masticatory muscles in the rabbit, the responses of muscle spindle units were recorded from the mesencephalic trigeminal nucleus (MesV) under halothane anesthesia during ramp-and-hold stretches. Three firing patterns, initial burst (IB) at the onset of the dynamic phase, negative adaptation (NA) at the end of the dynamic phase and firing during the release (FDR) phase, were observed during muscle stretch. IB was present at higher stretch velocities, FDR at lower stretch velocities. The velocity at which an IB or FDR was present was different from unit to unit. Because, within the range of the velocities of stretch tested, units with NA always showed NA and units without NA never did, all recorded units were divided into two groups on the basis of the existence of NA (NA(+) or NA(-) units). Response characteristics of the two groups were then compared. NA(+) units showed an IB more frequently and FDR less frequently than NA(-) units. NA(+) units had significantly higher dynamic responsiveness and discharge variability than NA(-) units. The conduction velocity of the afferents of NA(+) units was higher than that of NA(-) units. However, distributions of these measurements were not bimodal. These results suggest that NA is the useful criteria to classify the muscle spindle endings in the masticatory muscles in the rabbit under halothane anesthesia. PMID- 10375672 TI - Changes of somatosensory evoked potentials during writing with the dominant and non-dominant hands. AB - Since close attention and special effort are necessary to perform difficult unskilled movements, particular brain activities underlying such movements could be expected to take place in the primary sensori-motor cortices (SI and MI). In this study we focused on such activities by analyzing the difference in the somatosensory evoked potential (SEP) in presence to the electrical stimulation of the median nerve during writing using the dominant and non-dominant hands in twelve right-handed and eight left-handed normal subjects. By alternately stimulating the right and left median nerves during the writing with either hand, SEPs were recorded from both hemispheres. During the dominant hand writing, the middle latency SEP components, i.e., parietal P25 and N33 and frontal N30, were significantly attenuated only in the hemisphere contralateral to the writing hand, corresponding to the conventional gating effect. During the non-dominant hand writing, not only those components recorded from the hemisphere contralateral to the writing hand, but also those from the hemisphere ipsilateral to the writing hand were significantly attenuated. In addition, N20 in the hemisphere contralateral to the writing hand was also significantly attenuated. There was no significant difference in the attenuation between the right-handed and left-handed subjects. The results indicated that the specific interaction between the signals after electrical stimulation and the sensory cortical activities related to the writing using the non-dominant hand occurred in both hemispheres, while it was recognized only in the hemisphere contralateral to the writing hand during the dominant hand writing. We speculate that the somatosensory cortex was more activated and thus interacted with the applied stimulation during the unskilled movement of the non-dominant hand compared to the movement of the dominant hand. PMID- 10375673 TI - Calcium and increase excitability promote tolerance against anoxia in hippocampal slices. AB - We have previously demonstrated that anoxic preconditioning (APC) protects against a subsequent otherwise 'lethal' anoxic insult in hippocampal slices. Tested here are two hypotheses: (a) APC requires calcium to improve electrical recovery in hippocampal slices; and (b) mild excitation promotes preconditioning neuroprotection. Control hippocampal slices were given a single 'test' anoxic insult followed by reoxygenation. Experimental slices were preconditioned by three short anoxic insults of 1 min separated by 10 min of reoxygenation. At 30 min after the third 'conditioning' insult, slices underwent a 'test' anoxic insult [1 min of anoxic depolarization (AD)], and then slices were reoxygenated. Evoked potentials (EPs) were recorded throughout the experiment. In other slices, APC was emulated by inducing spreading depression (as determined by a negative DC shift) with KCL or by inducing increased neuronal excitability with the excitatory agent 8-cyclopentyl-1, 3-dipropylxanthine (DPCPX) (an adenosine A1 receptor blocker). 'Test' anoxic insults lasted 2 min of AD in these groups. To determine the role of calcium during APC, extracellular CaCl2 was decreased to 0.5 mM but only during the APC episodes ('test' anoxia, 1 min of AD). EP amplitudes recovered significantly better after anoxia in preconditioned slices, and in KCl- and DPCPX-treated slices (147.2+/-33.3, n=8, **p<0.01, 71.7+/-13.5, n=7, **p<0.01, and 117.8+/-37.3, n=5, ***p<0.001, respectively) compared to controls. Decreases in extracellular CaCl2 during APC blocked the recovery of EPs after 'test' anoxia (80.6+/-23.0, n=8). These data confirm that increases in excitability can emulate APC. These data also demonstrate that calcium influx during preconditioning is required for the induction of tolerance during APC. PMID- 10375674 TI - Circadian rhythm of AMPA receptor GluR2/3 subunit-immunoreactivity in the suprachiasmatic nuclei of Syrian hamster and effect of a light-dark cycle. AB - In mammals, the suprachiasmatic nuclei (SCN) of the hypothalamus are the site of the circadian clock that generates and coordinates many endogenous physiological and behavioral rhythms. SCN are normally entrained to light/dark (LD) cycle by direct retinal afferents using glutamate as neurotransmitter. N-Methyl-d aspartate (NMDA) and non-NMDA receptors are involved in photic entrainment of SCN. In rodents, the presence of three of the four known 2-amino-3-(3-hydroxy-( methylisoaxol-4-yl) propanoic acid (AMPA) receptor subunits has been demonstrated by in situ hybridization. This study analyzes the expression of GluR2/3 subunits in SCN of Syrian hamsters maintained under constant darkness (DD) or 12:12 LD cycle. In animals submitted to DD or LD, small immunoreactive neurons were located in the ventral and external latero-ventral parts of the rostral two thirds of the SCN and along the symmetrical plane. The number of intensely labeled neurons with or without long process(es) were counted at six circadian times (CTs) in three groups of animals maintained in DD and six nycthemeral (zeitgeber time, ZT) times in one group of hamsters submitted to LD. In DD, we observed significantly more GluR2/3 subunit-immunoreactive (GluR2/3-ir) neurons during the subjective day than during the subjective night, with minima at CT 19 CT 23. The LD cycle significantly reduced the number of immunoreactive neurons, lessened the differences between LD phases and depressed immunoreactivity at light transition, i.e., at ZT 11 and ZT 23. This study demonstrates for the first time by immunohistochemistry the existence of a circadian dynamic of the expression of AMPA receptor subunits in SCN of rodents and the effect of the LD cycle on this dynamic. PMID- 10375675 TI - Analyses of signal transduction cascades in rat pinealocytes reveal a switch in cholinergic signaling during postnatal development. AB - In the rat pineal gland, norepinephrine activates alpha1- and beta-adrenergic receptors and triggers melatonin production through an increase in the intracellular calcium concentration ([Ca2+]i) and stimulation of the cAMP/cAMP responsive element-binding protein (CREB) cascade. VIP and PACAP also elevate the intracellular cAMP level and promote melatonin formation. Finally, ACh antagonizes the norepinephrine-induced hormone synthesis via nicotinic acetylcholine receptors and subsequent activation of voltage-gated calcium channels. By immuno(cyto)chemical demonstration of phosphorylated CREB and calcium imaging we have investigated the temporal relationship between the maturation of these signaling pathways and the rhythmic onset of melatonin biosynthesis in developing rat pinealocytes. Norepinephrine-regulated calcium signaling and phosphorylation of CREB are already fully developed at birth, i.e., prior to ingrowth of the sympathetic innervation into the pineal parenchyma, and appear to develop in an innervation-independent manner. VIP- and PACAP-induced CREB phosphorylation is restricted to subpopulations of neonatal cells and thus also displays an adult pattern. Cholinergic calcium signaling exhibits a developmental switch within the first three postnatal weeks. In neonatal pinealocytes, acetylcholine elevates [Ca2+]i via muscarinic rather than nicotinic acetylcholine receptors. In the second postnatal week, pinealocytes gain responsiveness to nicotine and gradually lose responsiveness to muscarinic cholinergic stimuli. Voltage-gated calcium channels are absent in neonatal cells and develop during the first postnatal days. ACh-evoked cellular events may be diversified depending on the functional subclass of receptor that is present. The transient existence of muscarinic acetylcholine receptors and the subsequent switch to nicotinic receptors would permit ACh to elicit temporary effects in early pineal development. PMID- 10375676 TI - Medial septum mediates the increase in post-ictal behaviors and hippocampal gamma waves after an electrically induced seizure. AB - Hippocampal EEG and behavior of freely moving rats were studied before and after a hippocampal afterdischarge (AD), with or without reversible inactivation of the medial septum (MS) by muscimol. Muscimol suppressed the normal hippocampal EEG, including theta (5-10 Hz) and gamma (30-70 Hz) waves. After a hippocampal AD, hippocampal gamma waves were decreased for about 2 min and then increased at 3-10 min, while power of EEG of <30 Hz was decreased at 300 Torr) causes Ca2+-dependent photolabile excitation of chemosensors in the carotid body (CB). We previously proposed that the source of this Ca2+ was the [Ca2+]i stores because CO would react only intracellularly. However, influx of extracellular Ca2+ was not excluded. Now, using perfused rat CB (n=6) in the presence of normal extracellular [Ca2+] we show that chemosensory response to CO (pCO approximately 550 Torr) in normoxic (pO2 approximately 100 Torr) normocapnia (pCO2 approximately 30 Torr, pH approximately 7.4) is completely but reversibly inhibited by Cd2+ (200 microM), a voltage-gated Ca2+ channel blocker. Thus, extracellular Ca2+ is necessary for excitatory chemosensory response to high pCO. Cd2+ block occurs in spite of an enhanced [Ca2+]i rise. This shows that Ca2+ rise alone is unable to release neurotransmitter and to elicit a chemosensory response. Therefore, as a corollary, we conclude that Cd2+ blocks the Ca2+ flux that is needed for vesicle-membrane fusion for neurotransmitter release and neural discharge. PMID- 10375684 TI - The effect of age on expression of endogenous plasminogen activators after focal cerebral ischemia in mice. AB - We measured urokinase-type plasminogen activator (u-PA) and tissue-type plasminogen activator (t-PA) activity in the brain of 2-3 month old and 6-8 month old mice subjected to 4 h of middle cerebral artery (MCA) occlusion. t-PA activity was present in all non-ischemic and ischemic young mouse brain. In contrast, t-PA activity was present in 46.7% of non-ischemic middle aged mouse brain and in 44.4% of ischemic middle aged mouse brain. u-PA activity was present in all young and middle aged non-ischemic brains. PMID- 10375685 TI - No benefit of dietary restriction on disease onset or progression in amyotrophic lateral sclerosis Cu/Zn-superoxide dismutase mutant mice. AB - Amyotrophic lateral sclerosis (ALS) is a fatal disease in which spinal cord motor neurons degenerate resulting in progressive paralysis. Some cases of ALS are caused by mutations in the antioxidant enzyme Cu/Zn-superoxide dismutase (SOD). Transgenic mice expressing ALS-linked Cu/Zn-SOD mutations (SODMutM) exhibit a phenotype analogous to that of human ALS patients. Dietary restriction (DR) is a well-established means of extending lifespan in rodents. It may act by reducing levels of cellular oxidative stress. We therefore tested the hypothesis that DR will retard the development of the clinical phenotype and extend the lifespan of SODMutM. There was no significant difference in age of disease onset in mice placed on a DR regimen beginning at 6 weeks of age compared to mice fed ad libitum, and disease duration was shortened indicating that DR accelerates the clinical course. Histological analyses indicated a similar extent of lower motor neuron degeneration in SODMutM maintained on DR or ad libitum diets. We conclude that a dietary manipulation known to extend lifespan has no beneficial effect in an animal model of familial ALS. PMID- 10375686 TI - Ultrastructural localization of hippocampal TNF-alpha immunoreactive cells in rats following transient global ischemia. AB - Using a polyclonal antibody against rat TNF-alpha, we have identified specific intracellular immunoreactive sites in hippocampal pyramidal cells, astroglia, and in microglia within 72 h after a period of ischemia. Electron opaque immunoreactive products in pyramidal cells were found mainly in somata and dendrites. Astrocytes and microglia were nearly devoid of such complexes. These findings demonstrate the presence of TNF-alpha in hippocampal neurons and its enhancement by ischemic stress. PMID- 10375687 TI - Alteration of iron homeostasis following chronic exposure to manganese in rats. AB - Recent studies suggest that manganese-induced neurodegenerative toxicity may be partly due to its action on aconitase, which participates in cellular iron regulation and mitochondrial energy production. This study was performed to investigate whether chronic manganese exposure in rats influenced the homeostasis of iron in blood and cerebrospinal fluid (CSF). Groups of 8-10 rats received intraperitoneal injections of MnCl2 at the dose of 6 mg Mn/kg/day or equal volume of saline for 30 days. Concentrations of manganese and iron in plasma and CSF were determined by atomic absorption spectrophotometry. Rats exposed to manganese showed a greatly elevated manganese concentration in both plasma and CSF. The magnitude of increase in CSF manganese (11-fold) was equivalent to that of plasma (10-fold). Chronic manganese exposure resulted in a 32% decrease in plasma iron (p<0.01) and no changes in plasma total iron binding capacity (TIBC). However, it increased CSF iron by 3-fold as compared to the controls (p<0.01). Northern blot analyses of whole brain homogenates revealed a 34% increase in the expression of glutamine synthetase (p<0.05) with unchanged metallothionein-I in manganese intoxicated rats. When the cultured choroidal epithelial cells derived from rat choroid plexus were incubated with MnCl2 (100 microM) for four days, the expression of transferrin receptor mRNA appeared to exceed by 50% that of control (p<0.002). The results indicate that chronic manganese exposure alters iron homeostasis possibly by expediting unidirectional influx of iron from the systemic circulation to cerebral compartment. The action appears likely to be mediated by manganese-facilitated iron transport at brain barrier systems. PMID- 10375688 TI - Dopamine depletion in the medial prefrontal cortex induces sensitized-like behavioral and neurochemical responses to cocaine. AB - It has been postulated that behavioral sensitization to cocaine is associated with an attenuation of cocaine-induced dopamine (DA) transmission in the medial prefrontal cortex (mPFC). Hence, experiments were designed to examine the effects of chemically-induced cortical DA depletion on the acute behavioral and neurochemical responses to cocaine. One week following two bilateral 6 hydroxydopamine (6-OHDA) injections into the mPFC, animals received injections of cocaine (7.5, 15 or 30 mg/kg, i.p.) or saline (1 ml/kg, i.p.) in a randomized fashion with a minimum 3 day intertrial interval. Cocaine produced a dose dependent increase in motor activity which was significantly enhanced in animals depleted (mean of 76%) of dopamine in the mPFC. Likewise, 6-OHDA lesions of the mPFC produced a significant enhancement of cocaine-induced DA transmission in the nucleus accumbens (NAC) as estimated by in vivo microdialysis. These data indicate a permissive involvement of cortical DA in mediating behavioral and neurochemical responses to cocaine, as well as confirm the ability of the mPFC to influence subcortical structures in response to an acute injection of cocaine. Collectively, the present findings suggest that alterations in cortical DA transmission may be a neural substrate mediating the development of sensitization to cocaine, and thus, may contribute to the addictive properties of cocaine. PMID- 10375689 TI - Regulation of protein phosphorylation in astrocyte cultures by external calcium ions: specific effects on the phosphorylation of glial fibrillary acidic protein (GFAP), vimentin and heat shock protein 27 (HSP27). AB - The effect of external Ca2+ ([Ca2+]e) on the incorporation of [32P] into total protein, cytoskeletal proteins and the heat shock protein HSP27, was studied in primary cultures of astrocytes from the rat hippocampus. Zero [Ca2+]e increased total 32P-incorporation into astrocyte protein and when this was normalized to 100%, incorporation was significantly increased into glial fibrillary acidic protein (GFAP), vimentin (VIM) and HSP27. The difference in total 32P incorporation between zero [Ca2+]e and 1 mM [Ca2+]e was reversed by incubation of the cells with the protein phosphatase inhibitor okadaic acid in the range 1-10 nM; higher concentrations of okadaic acid (50-100 nM) further increased total 32P incorporation. In zero [Ca2+]e the non-specific channel blocker Co2+ (1 mM) decreased total 32P-incorporation by approximately 30%. The results were compared with a previous study [S.T. Wofchuk, R. Rodnight, Age-dependent changes in the regulation by external calcium ions of the phosphorylation of glial fibrillary acidic protein in slices of rat hippocampus, Dev. Brain Res. 85 (1995) 181-186] in which it was shown that in immature hippocampal slices zero [Ca2+]e compared with 1 mM [Ca2+]e increased 32P-incorporation into GFAP without changing total incorporation. The difference between the results for total 32P-incorporation obtained in cultured astrocytes and immature brain tissue was found to be related to the concentration of [Ca2+]e in the medium since in slices concentrations of [Ca2+]e higher than 1 mM progressively decreased total incorporation. The difference may reflect a higher Ca2+-permeability of the plasma membrane in cultured astrocytes and/or to the complex structure of the slice tissue. In two dimensional electrophoresis HSP27, in contrast to GFAP and VIM, was separated into 3 immunodetectable isoforms only two of which were normally phosphorylated. After labelling in the presence of okadaic acid both immunodetectable and phosphorylated HSP27 focussed as a single polypeptide. Phorbol dibutyrate (1 microM) and zero [Ca2+]e stimulated the phosphorylation of both isoforms, but in the case of zero [Ca2+]e the effect on the more acidic isoform was proportionally greater. PMID- 10375690 TI - Effects of handling on extracellular levels of glutamate and other amino acids in various areas of the brain measured by microdialysis. AB - Upon a physiological and pharmacological challenge, the responsiveness of extracellular glutamate levels in the prefrontal cortex, ventral tegmental area and locus coeruleus were studied using microdialysis. A 10-min handling period was used as a mild stressful stimulus. In all three brain areas, handling induced an immediate and short-lasting increase in glutamate levels, but the responses were highly variable. Only in the ventral tegmental area and the locus coeruleus, but not in the prefrontal cortex, the increases were significantly different from basal values. In rats with relatively low basal glutamate levels, both in the ventral tegmental area and locus coeruleus, handling had a more pronounced effect on glutamate levels than in rats with high basal levels, although in some rats with relatively low basal levels of glutamate, handling had hardly any effect. Potassium stimulation also induced variable responses in all three brain areas. Again, relatively low basal glutamate levels were more responsive to the stimulation than higher basal values, although there appeared to be a lower limit. These data suggest that relatively high basal levels contain sources of glutamate that mask the neuronal pool of glutamate and are therefore less responsive to physiological or pharmacological stimulation. However, this interpretation was questioned by the findings that basal levels and handling induced increases in glutamate levels were found to be (partly) TTX-independent. As carrier-mediated release as a possible non-exocytotic release mechanism has only been described in vivo under pathological conditions, it seems plausible to ascribe TTX-independent glutamate increases to aspecific, non-neuronal processes. This interpretation was further supported by the observation that in all three brain areas, other amino acids, i.e., aspartate, taurine, glutamine, serine, alanine and glycine also increased upon handling in a very similar way as glutamate did. Thus, these results question a direct correlation between stimulated extracellular glutamate levels induced by handling and measured by microdialysis and glutamatergic neurotransmission. PMID- 10375691 TI - Regional changes in the expression of neurotrophic factors and their receptors following acute traumatic brain injury in the adult rat brain. AB - We have analyzed the effect of severe traumatic brain injury (TBI) on the levels of mRNA expression of neurotrophic factors (NTFs): brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), ciliary neurotrophic factor (CNTF) and their respective receptors: trkB, trkA and CNTFRalpha. The expression was examined in the region of the lesion as well as a region remote from the lesion at 12, 24, and 36 h following the injury. Our data suggest that after the brain injury, the expression of NGF and BDNF mRNAs were early, transiently and significantly upregulated while that of CNTF was a slow and less amplified response in both areas of the brain. We also found that trkA mRNA expression was only upregulated significantly in the remote area; trkB mRNA showed no significant change in either area except an upregulation at 12 h in the remote area. CNTFRalpha was downregulated significantly by 24-36 h in the lesion area and by 24 h in the remote area. These changes suggest that TBI regulates the expression of NTFs and their receptors. These alterations in expression may be involved in modulating the neuronal response after brain injury. PMID- 10375692 TI - Scatter factor/hepatocyte growth factor gene transfer increases rat blood-glioma barrier permeability. AB - Malignant gliomas are associated with a dysfunctional blood-tumor barrier (BTB) that causes substantial morbidity. Scatter factor/hepatocyte growth factor (SF/HGF) is a multifunctional growth factor that correlates with glioma malignancy and has several biological properties that suggest a role in enhancing blood-glioma barrier permeability. In this study, we examined the effects of glioma cell SF/HGF expression on BTB permeability to horseradish peroxidase (HRP). Fischer 344 rats bearing intrastriatal 9L tumors engineered to secrete SF/HGF (9L-SF) and SF/HGF-negative control tumors (9L-neo) received intracardiac injections of HRP and were rapidly decapitated. Densitometric analysis of brain sections reacted with diaminobenzidine showed significantly greater extravascular HRP surrounding SF/HGF-secreting tumors than 9L-neo tumors of comparable size (p<0.05). HRP enzymatic activity associated with striata containing SF/HGF expressing tumors was 1. 6-fold greater than that of striata containing control tumors (p<0. 05). Northern analysis showed that expression of vascular endothelial growth factor/vascular permeability factor (VEGF/VPF) did not differ between 9L-neo and 9L-SF tumors. These data demonstrate that SF/HGF expression by intracerebral glial tumors can enhance BTB permeability independent of changes in VEGF/VPF expression. PMID- 10375693 TI - Middle cerebral artery occlusion in the mouse by intraluminal suture coated with poly-L-lysine: neurological and histological validation. AB - The present study was conducted to validate a modified method of temporary focal cerebral ischemia in the mouse; neurobehavioral function and histopathological infarction were quantitated following various periods of middle cerebral artery occlusion (MCAo). Male C57BL/6 mice were anesthetized with 3% halothane in a mixture of 30%O2/70%N2O delivered by face mask and were subjected to 30- to 180 min of temporary middle cerebral artery occlusion (MCAo) by an intraluminal suture coated with poly-l-lysine. Twenty-eight of 40 mice showed an initial high grade neurological deficit (30-min MCAo, n=7; 60-min, n=8; 120-min, n=8; 180-min, n=5) when examined during MCAo; these were used for subsequent study. One day after MCAo, behavioral function was re-evaluated, and brains were perfusion-fixed and infarct volumes were measured. The initial neurological deficit improved at 24 h in mice with 30- or 60-min of prior MCAo but tended to persist in mice with 120- or 180-min insults. Following each duration of ischemia, mice exhibited ipsilateral infarcts. Small, inconsistent predominantly subcortical infarcts were present after 30-min MCAo, while longer occlusion periods gave rise to consistent foci of subcortical infarction involving striatum, septum, thalamus, and hippocampus, as well as areas of frontoparietal cortical infarction. The major advantages of the improved intraluminal MCAo model reported here, incorporating sutures coated with poly-l-lysine, include: a 100% incidence of infarction of predictable location and size in mice having an initial neurological deficit. Periods of 60- to 180-min MCA occlusion in this model yield sufficiently reproducible sequelae to permit the effects of various therapeutic agents on neurological outcome and size of infarction to be readily studied. PMID- 10375694 TI - Corticostriatal and corticosubthalamic input zones from the presupplementary motor area in the macaque monkey: comparison with the input zones from the supplementary motor area. AB - The presupplementary motor area (pre-SMA) is a cortical motor-related area which lies in the medial wall of the frontal lobe, immediately anterior to the supplementary motor area (SMA). This area has been considered to participate in the control of complex forelimb movements in a way different from the SMA. In an attempt to analyze the patterns of projections from the pre-SMA to the basal ganglia, we examined the distributions of pre-SMA inputs in the striatum and the subthalamic nucleus and compared them with the SMA input distributions. To detect morphologically the terminal fields from the pre-SMA and the forelimb region of the SMA, anterograde tracers were injected into such areas that had been identified electrophysiologically in the macaque monkey. Corticostriatal inputs from the pre-SMA were distributed mainly in the striatal cell bridges connecting the rostral aspects of the caudate nucleus and the putamen, as well as in their neighboring striatal portions. These input zones were located, with no substantial overlap, rostral to corticostriatal input zones from the SMA forelimb region. Corticosubthalamic input zones from the pre-SMA were almost localized in the medial aspect of the nucleus, where corticosubthalamic inputs from the SMA forelimb region were also distributed predominantly. However, the major terminal fields from the pre-SMA were centered ventrally to those from the SMA. The present results indicate that the corticostriatal and corticosubthalamic input zones from the pre-SMA appear to be segregated from the SMA-derived input zones. This implies the possibility of parallel processing of motor information from the pre-SMA and SMA in the cortico-basal ganglia circuit. PMID- 10375695 TI - Acetaldehyde cytotoxicity in cultured rat astrocytes. AB - The effect of acetaldehyde on astrocytes have been investigated because not only do they play an important role in brain maturation but also recent reports have shown their delayed proliferation following both 'in vivo' and 'in vitro' ethanol exposure. Biochemical parameters related to apoptotic and necrotic processes were examined in primary cultures of rat astrocytes exposed for 4 days to acetaldehyde generated from ethanol by co-cultured alcohol dehydrogenase-transfected Chinese hamster ovary cells. Acetaldehyde levels in the culture media attained concentrations of approximately 450 microM. To study ethanol effects, alcohol oxidation was inhibited by 4-methylpyrazole (an inhibitor of alcohol dehydrogenase). Acetaldehyde but not ethanol increased intracellular calcium levels by 155%. Moreover, significant DNA fragmentation was detected using a random oligonucleotide primed synthesis assay, by flow cytometry and when using agar gel electrophoresis. Transglutaminase activity was elevated in the cells treated with acetaldehyde but when acetaldehyde formation was inhibited by 4 methylpyrazole the enzyme activity was unaffected. Nitrate levels in the culture media were unchanged. Additionally, microscopic examination of cell nuclei revealed chromatin condensation in astrocytes exposed to acetaldehyde. It can be concluded, that in 'in vitro' acetaldehyde exposed rat astrocytes apoptotic pathways are activated. PMID- 10375697 TI - Behavioral effects of cocaine on a transgenic mouse model of cortical-limbic compulsion. AB - We previously created a transgenic mouse model of cortical-limbic induced compulsions in which dopamine D1 receptor-expressing (D1+) neurons in restricted regional subsets of the cortex and amygdala express a neuropotentiating cholera toxin (CT) transgene. These 'D1CT' mice engage in complex behavioral abnormalities uniquely resembling human compulsions, such as non-aggressive biting of cagemates during grooming, repeated leaping and episodes of perseverance of any and all normal behaviors. Because both compulsions and cocaine-induced behaviors may represent forms of psychomotor activation that have a shared or overlapping neurological basis, we have examined the behavioral response of these 'compulsive' mice to cocaine. In both control and D1CT mice, cocaine increased the amount of time spent engaged in typical cocaine-dependent stereotypies such as locomotion, sniffing, or gnawing, while the remainder of behaviors within their normally complete behavioral repertoires decreased. Cocaine also decreased, rather than facilitated, the incidence of D1CT transgene induced compulsion-like behaviors such as repeated leaping and perseverance of any and all normal behaviors. The indistinguishable cocaine responses of D1CT and normal mice, as well as the masking (rather than potentiation) of D1CT mouse compulsion-like behaviors by cocaine, suggests that cortical-limbic induced compulsions are significantly different in their origin or circuitry from cocaine induced stereotyped behaviors. Specifically, these data suggest that the motor circuits stimulated in compulsions represent only a subset of the parallel circuits stimulated by cocaine. These data are, thus, consistent with the hypothesis that topographically restricted subsets of parallel cortical-striatal thalamic loops induce different types of compulsive behaviors. PMID- 10375698 TI - Neuronal activity in the cerebellar interpositus and lateral pontine nuclei during inhibitory classical conditioning of the eyeblink response. AB - Single-unit neuronal activity was recorded from the cerebellar interpositus nucleus and lateral pontine nuclei during conditioned inhibition of the eyeblink response in rats. Conditioned inhibition training sessions included 100 trials/day for 12 days. During each training session, the rats were given 50 presentations of a tone conditioned stimulus (CS) that was paired with a brief periocular shock unconditioned stimulus (US). They were also given 50 presentations of a compound stimulus that included the tone-CS and a light-CS. The compound-CS was not paired with the US. The two types of trials were mixed throughout the session and presented in an irregular sequence. This training procedure resulted in significant inhibition of the eyeblink response during the compound-CS. Neurons in the interpositus and lateral pontine nuclei exhibited significantly less activity during the compound-CS relative to the tone-CS. The suppression of cerebellar and pontine learning-related neuronal activity during the inhibitory CS may be critical for inhibiting the conditioned eyeblink response. PMID- 10375696 TI - Neuronal localization of a novel mosaic apolipoprotein E receptor, LR11, in rat and human brain. AB - A new type of mosaic protein was recently discovered as a new member of the low density lipoprotein receptor (LDLR) family, designated as LR11. The predominant expression of LR11 transcripts in brain tissue and the presence of elements found in neural adhesion molecules suggested a function(s) in the central nervous system (CNS). In order to gain insight about this complex receptor in the CNS, we raised a rabbit polyclonal antibody and examined immunohistochemically rat and human brain tissue. A strong LR11 immunoreactivity was found to be localized mainly in neurons throughout the brain in both species. A detailed mapping in the rat brain showed a distribution of LR11 immunoreactivity in a widespread population of neurons, though the intensity varied between different locations. The most prominent immunoreactivity was observed in neurons of the hippocampus, some nuclei of brain stem and Purkinje cells, whereas neurons of the thalamus and the hypothalamus showed weak staining. Uniquely, the single LR11 immunoreactive cytoplasmic puncta were observed in the proximity of apical dendrites, most conspicuously in the pyramidal neurons of hippocampus. In the human brain, one to four immunoreactive puncta were seen within individual neurons. The neuronal localization of LR11 and its unique association of cytoplasmic structure, presumably botrysome, may suggest the roles of LR11 in both the lipoprotein metabolism and intracellular trafficking in certain neuronal population of the CNS. PMID- 10375699 TI - Metabotropic glutamate receptor modulation of excitotoxicity in the neostriatum: role of calcium channels. AB - We have previously shown that metabotropic glutamate receptor (mGluR) activation can attenuate N-methyl-d-aspartate (NMDA)-induced excitotoxic injury in the neostriatum both in vivo and in vitro. Our earlier studies made use of the non subtype selective mGluR agonist 1-amino-cyclopentane-1,3-dicarboxylic acid (tACPD). In the present study, we extended these observations by identifying the subtype of mGluR involved. Using selective mGluR agonists, we provide evidence that the Group II mGluRs are responsible for inhibition of NMDA excitotoxicity in the neostriatum. In addition, we provide evidence that the inhibitory effects of tACPD on excitotoxicity are dependent upon calcium influx as they are blocked by a low calcium solution as well as the broad-spectrum calcium channel blocker cadmium. The tACPD-induced attenuation was also blocked by omega-conotoxin GVIA suggesting participation of N-type calcium channels. Whole cell voltage clamp recordings were made to directly determine the effects of mGluRs on voltage-gated calcium channels in neostriatal neurons. As predicted, both tACPD and the Group II agonist 3C4HPG inhibited calcium currents in neostriatal neurons. Again this effect was blocked by omega-conotoxin GVIA. Overall the results suggest that mGluR regulation of voltage-gated calcium channels can limit NMDA toxicity in the neostriatum. PMID- 10375700 TI - Disinhibition of lower midbrain neurons enhances non-shivering thermogenesis in anesthetized rats. AB - The hypothesis that the lower midbrain, specifically in and around the retrorubral field (RRF) and/or rubrospinal tract (rs), contains a tonic inhibitory mechanism on non-shivering thermogenesis (NST) was examined in urethane-anesthetized rats. It has been proposed that removal of the tonic inhibitory mechanism in the lower midbrain causes body temperature increase through disinhibition-induced activation of the central sympathetic nervous system. The present experiments were carried out to examine whether and where the proposed midbrain region contains cell bodies that tonically inhibit the NST, and if so, whether they receive any influence from the hypothalamus. Male Wistar rats were anesthetized with urethane (1. 0-1.2 g/kg, i.p.), and various agents were microinjected into the RRF and rs areas of one side before and after knife-cut in the other side of the lower midbrain or isolation of the hypothalamus from the midbrain. Changes in interscapular brown adipose tissue (IBAT), rectum, and tail skin temperatures were monitored. RESULTS: (1) unilateral midbrain procaine increased IBAT and rectal temperatures only after un-injected side of the midbrain had been pre-transected. (2) Effective midbrain sites for procaine to increase IBAT and rectal temperatures was laterally extended. (3) Tetrodotoxin microinjected into the midbrain site where procaine increased IBAT and rectal temperatures also raised both temperatures. (4) l-glutamate decreased IBAT and rectal temperatures when microinjected into one of the most inner midbrain area of procaine-sensitive sites without affecting tail skin temperature. (5) Isolation of the hypothalamus from the lower midbrain did not affect midbrain procaine-induced IBAT and rectal temperature increases. These results suggest that neurons that tonically inhibit the NST are located in the area close to the midline adjacent to the RRF and rs, and that the integrity of the neurons to tonically inhibit the NST is not affected by disconnecting the hypothalamus from the midbrain. PMID- 10375701 TI - The effect of intrathecal selective agonists of Y1 and Y2 neuropeptide Y receptors on the flexor reflex in normal and axotomized rats. AB - We have examined the effects of intrathecal (i.t.) administration of [Leu31,Pro34]-neuropeptide Y (NPY) or NPY-(13-36), selective agonists of NPY Y1 or Y2 receptors, respectively, on the excitability of the flexor reflex in normal rats and after unilateral transection of the sciatic nerve. In rats with intact and sectioned sciatic nerves, i.t. [Leu31,Pro34]-NPY induced a similar biphasic effect on the flexor reflex with facilitation at low doses and facilitation followed by depression at high doses. In contrast, i.t. NPY-(13-36) only facilitated the flexor reflex in normal rats, and at high dose it caused ongoing discharges in the electromyogram. NPY-(13-36) caused dose-dependent depression of the flexor reflex in rats after sciatic nerve transection, in addition to its facilitatory effect. Topical application of [Leu31,Pro34]-NPY or NPY-(13-36) caused a moderate and brief reduction in spinal cord blood flow. No difference was noted between the vasoconstrictive effect of [Leu31,Pro34]-NPY and NPY-(13 36). It is suggested that activation of Y1 receptors may be primarily responsible for the reflex depressive effect of i.t. neuropeptide Y in rats with intact sciatic nerves, whereas both Y1 and Y2 receptors may be involved in mediating the depressive effect of NPY after axotomy. PMID- 10375702 TI - Degeneration and gliosis in rat retina and central nervous system following 3,3' iminodipropionitrile exposure. AB - 3,3'-Iminodipropionitrile (IDPN) exposure causes a neurofilamentous axonopathy and olfactory, audiovestibular and visual toxicity. Many events relevant to these effects and the neurotoxic properties of nitriles as a class remain to be elucidated. We characterized the gliosis associated with the IDPN-induced retinal degeneration in comparison to other effects on the visual and central nervous systems. Gliosis was quantified using an ELISA for the intermediate filament protein, glial fibrillary acidic protein (GFAP). IDPN (0-400 mg kg-1 day-1x3 days, i.p.) caused corneal opacity and dose- and time-dependent increases in retinal GFAP, up to 26-28 fold of control values at 4 weeks post-exposure; a second peak occurred at 16 weeks. In contrast, GFAP peaked at 1 week in olfactory bulbs (OB), cingulate cortex and hippocampus. Cerebellum and striatum showed no gliosis. Retinal dopamine decreased within 2 weeks. Delayed GFAP increases occurred in superior and inferior colliculi. Retina and superior colliculi also showed increased [3H]PK-11195 binding. Histological analysis demonstrated progressive degeneration and gliosis in retina and colliculi. Taken together, the data indicate that primary and secondary degenerative events occur in the retina, and that this retinal degeneration induces GFAP increases in retina and superior colliculus. In addition, GFAP assays demonstrated that the retinal toxicity of IDPN is enhanced by CCl4 hepatotoxicity and blocked by methimazole inhibition of flavin-mono-oxygenases, similarly to its ototoxicity. GFAP assays also indicated that neither vestibulotoxic doses of crotononitrile nor olfatotoxic doses of dichlobenil damage the retina. The data support the use of GFAP assays for assessing the retinal toxicity of IDPN and other nitriles. PMID- 10375703 TI - Actions of 8-bromo-cyclic-GMP on neurones in the rat thalamus in vivo and in vitro. AB - The diffusible intercellular messenger nitric oxide may have a modulatory role in the thalamus and this action may be mediated via activation of soluble guanylate cyclase. In order to investigate this possibility, we applied the cyclic-GMP analogue 8-Bromo-cyclic-GMP (8-Br-cGMP) onto neurones in the ventrobasal and lateral geniculate nuclei of the thalamus in anaesthetised rats, and compared its effects with those of a nitric oxide donor. 8-Br-cGMP enhanced the responses of neurones to iontophoretically applied NMDA and AMPA. Furthermore, somatosensory and visual responses of ventrobasal and lateral geniculate neurones were enhanced to 274+/-76% and 217+/-69% of control values, respectively. These effects were similar to those seen with nitric oxide donors in this study and previous work from this laboratory. When applied to thalamic neurones in an in vitro slice preparation, 8-Br-cGMP caused a membrane depolarisation associated with a decrease in input resistance. These findings indicate that activation of guanylate cyclase can cause a membrane depolarisation of thalamic neurones in vitro, and that this effect is sufficient to enhance action responses to ionotropic glutamate receptor stimulation via either exogenous agonists or sensory stimulation. PMID- 10375704 TI - Role of intracellular calcium in thermal allodynia and hyperalgesia in diabetic mice. AB - We examined the involvement of cytosolic calcium on thermal hyperalgesia and allodynia seen in diabetic mice. Tail-flick latencies at source voltages of a 50 W projection bulb to 35 and 50 V in diabetic mice were significantly shorter than those in non-diabetic mice. Tail-flick latencies at 35 and 50 V in diabetic mice were increased by pretreatment with ryanodine, which blocks Ca2+ release from Ca2+/caffeine-sensitive microsomal pools. On the other hand, intrathecal (i.t.) pretreatment with thapsigargin, which inhibits Ca2+ uptake into the inositol 1,4,5-trisphosphate-sensitive microsomal Ca2+ pool, decreased tail-flick latencies at 35 and 50 V in non-diabetic mice. Thus, it seems likely that thermal allodynia and hyperalgesia in diabetic mice may be due, in part, to the enhancement of intracellular calcium level in the spinal cord. PMID- 10375705 TI - Modification of the effect of diazepam on the propofol-induced loss of the righting reflex in mice by diabetes. AB - The effect of diabetes on the effect of diazepam on the propofol-induced loss of the righting reflex was investigated. There was no significant difference in the duration of the propofol-induced loss of the righting reflex between non-diabetic and diabetic mice. Diazepam increased the duration of the propofol-induced loss of the righting reflex in both diabetic and non-diabetic mice. The diazepam induced enhancement of the effect of propofol was significantly lower in diabetic mice than that in non-diabetic mice. These effects were antagonized by the pretreatment with flumazenil. Pretreatment with FG7142, a benzodiazepine receptor inverse agonist, attenuated the duration of the propofol-induced loss of the righting reflex in non-diabetic mice, but not in diabetic mice. These results suggest that the attenuation of the diazepam-induced enhancement of the duration of the propofol-induced loss of the righting reflex in diabetic mice may be due to the dysfunction of benzodiazepine receptors. PMID- 10375706 TI - Elevated neuropeptide Y and corticotropin-releasing factor in the brain of a novel epileptic mutant rat: Noda epileptic rat. AB - Noda epileptic rat (NER) is a new epileptic rat strain, which was developed by inbreeding rats with spontaneous tonic-clonic seizures in a stock of Crj:Wistar. In the present study, possible changes of two neuropeptides, neuropeptide Y (NPY) and corticotropin-releasing factor (CRF), in the brains of NER were investigated. Increased contents of immunoreactive (IR) NPY were found in the striatum and amygdala of 8-week NERs with partial seizure, while these changes extended to the limbic region including hippocampus in 16-week NERs with fully developed generalized tonic-clonic seizure. IR-CRF were elevated only in the entorhinal and pyriform cortex of both 8-week and 16-week NERs. Generalized tonic-clonic seizure in NERs induced a transient increase of NPY mRNA in the granular layer of dentate gyrus. These results suggest that NPY metabolism in the limbic brain contributes to the seizure susceptibility in this model of epilepsy. PMID- 10375707 TI - In vivo microdialysis measures of extracellular serotonin in the rat hippocampus during sleep-wakefulness. AB - We investigated extracellular 5-hydroxytryptamine (5-HT) levels in rat hippocampus during different stages of the sleep-waking cycle using in vivo microdialysis. The extracellular 5-HT level was highest in active waking (AW) and, when compared to AW, 5-HT level was progressively lower in quiet waking (QW; 78%), quiet sleep (QS; 50%) and REM (which we termed active sleep (AS); 40%). Functional implications of AS related-decreased 5-HT in the hippocampus are discussed. PMID- 10375708 TI - Complement regulators C1 inhibitor and CD59 do not significantly inhibit complement activation in Alzheimer disease. AB - Proteins characteristic of activated complement are associated with Alzheimer disease (AD) lesions. The classical complement pathway can be activated only when the influence of such endogenous regulators as C1-inhibitor (C1-inh) and CD59 are overcome. We used the techniques of reverse transcriptase-polymerase chain reaction and Western blotting to assess the mRNA and protein levels of C1-inh and CD59 in AD and control brains in comparison with levels of the complement components with which they interact. The inhibitors were only slightly upregulated and then only in heavily affected areas of AD brain such as the entorhinal cortex, hippocampus, midtemporal gyrus and midfrontal gyrus. The ratio of AD to control mRNAs in these four areas was 1.17 for C1-inh and 1.12 for CD59, compared to 3.06 for C1r, 2.67 for C1s, 2.35 for C5, 2.56 for C6, 2.42 for C7, 5. 08 for C8 and 16.3 for C9. Peripheral organ expression of C1-inh and CD59 mRNAs was no different in AD than controls but was slightly upregulated in infarcted heart tissue. Again, the increase was small compared with that of the competitive complement components. These data indicate that the forces which upregulate and activate complement in AD and myocardial infarction are not effectively suppressed by the endogenous regulators, C1-inh and CD59. PMID- 10375709 TI - Monoamine oxidase in the intermediolateral nucleus of the thoracic spinal cord of the rat. A histochemical study. AB - We examined monoamine oxidase (MAO) activity in the intermediolateral nucleus (IML) of the rat thoracic spinal cord by histochemistry with tyramine as a common substrate for both MAO types A and B. Light microscopy showed MAO activity in neuronal cell bodies, processes, and varicosities. Electron microscopic examination showed both MAO-positive and -negative neuronal cell bodies. In the stained cell bodies, histochemical reaction products were localized in the cytoplasm showing a selective association with mitochondrial outer membranes. MAO positive axon terminals were often found in contact with MAO-negative neurons but only occasionally with MAO-positive neurons. MAO histochemistry in the IML was also performed using serotonin (a MAO type A preferential substrate) and beta phenylethylamine (a MAO type B preferential substrate). Light microscopy identified MAO activity for serotonin in a plexus of varicosities but not in any neuronal cell bodies. The activity for beta-phenylethylamine was detected frequently in neuronal cell bodies but rarely in varicosities. Our findings indicate that two groups of IML neurons can be chemically distinguished, one contains MAO type B while the other lacks both MAO types A and B. In addition, many axon terminals contain MAO type A but only a few fibers include MAO type B in the IML. PMID- 10375710 TI - Aging and neuropathic pain. AB - Although chronic neuropathic pain disorders are more prevalent in the senescent population, little is known about how the aging process alters the thermal hyperalgesic sensitivity to peripheral nerve injury. In this study, neuropathic pain was induced in young, mature and aged FBNF1 hybrid rats via unilateral ligation of the left sciatic nerve. The extent to which the aging process affects the thermal hyperalgesic responsiveness of these animals was investigated. The results demonstrate that the aging process differentially alters nociceptive processing. PMID- 10375711 TI - Diurnal modulation of long-term potentiation in the hamster hippocampal slice. AB - Long-term potentiation (LTP) was examined in hippocampal slices from Syrian hamsters entrained to a LD 14:10 cycle. Population spike (PS) amplitudes from CA1 pyramidal cells were measured before (control) and after tetanizing the Schaffer/collateral commissural pathway. Slices from animals sacrificed during the day, between zeitgeber time (ZT) 0430 and 0530, were incubated, and then tetanized between ZT 1340 and 1930, where ZT=0 denotes lights on. Slices from animals sacrificed during the night, between ZT 1830 and 1930, were incubated, and tetanized between ZT 0030 and 0410. LTP, a sustained increase in PS amplitude following tetanus, was evoked in both groups. PS amplitude increased by 102.7+/ 20.3% in animals sacrificed during the day and by 48.0+/-7.5% in animals sacrificed during the night (p<0.05). Thus hamster slices prepared during the day show more robust LTP (a doubling of PS amplitude), a difference persisting in slices incubated for several hours. PMID- 10375712 TI - Lack of topographical organisation of the corticospinal tract in the cervical spinal cord of the adult rat. AB - The fluorescent dextran, Fluororuby, was injected into either the hindlimb or forelimb sensorimotor cortex to label corticospinal axons projecting to corresponding regions of spinal cord. In the rostral cervical spinal cord, labelled axons projecting to the lumbar or cervical grey matter were intermingled randomly. We conclude that there is no topographical organisation of the corticospinal tract in the cervical spinal cord. PMID- 10375713 TI - High-threshold calcium channel activity in rat hippocampal neurones during hypoxia. AB - Whole-cell patch clamp recordings in combination with direct control and measurements of O2 tension (pO2) in bath solution were used to determine the sensitivity of Ca2+ channels of cultured hippocampal neurones to hypoxia in glucose free solution. In all tested neurones, a lowering of pO2 to 4/50 mmHg did not induce changes either in magnitude, kinetics or voltage-current relations of total Ca2+ currents, which composed mainly from two types, L-type (64%) and N type (31%) components. Hypoxia only induced a delay of Ca2+ current run-down about 27.5% and 39% at 50 and 4 mmHg pO2 respectively that presumably depended on changes in cytoplasmic channel-modulatory metabolites. The obtained results demonstrate that Ca2+ channel molecules in cultured hippocampal neurones are themselves insensitive to short-lasting (10-20 min) oxygen and glucose deprivation, and that they are not a principal target for hypoxic influences on hippocampal function. PMID- 10375714 TI - The staff faces behind the work of the society: An introduction PMID- 10375715 TI - The faceless messengers. PMID- 10375716 TI - Predictive value of pulmonary venous flow patterns in detecting mitral regurgitation and left ventricular abnormalities. AB - OBJECTIVE: To determine whether abnormalities in pulmonary venous flow (PVF) patterns detected by transesophageal echocardiography (TEE) correlate with the severity of mitral regurgitation (MR) or the presence of left ventricular (LV) abnormalities, and to demonstrate whether a normal PVF pattern predicts the absence of structural heart disease. DESIGN: Review of all TEEs performed at a tertiary care cardiac hospital over a four-month period. PATIENTS: Among 195 studies, 100 fulfilled the inclusion criteria. RESULTS: PVF was categorized into three patterns, which have been described previously. A normal PVF pattern predicted the absence of clinically significant MR with a high degree of certainty (positive predictive value [PPV] 98%). However, it did not predict the absence of structural cardiac disease (PPV 64%). A PVF pattern that showed systolic flow reversal was strongly predictive of the presence of significant MR (sensitivity 86%, specificity 100%, PPV 100%). The frequency of significant MR in this group was much higher than in patients with normal PVF (12 of 12 versus one of 66, P<0.0001). Patients with a blunted PVF pattern were more likely than patients with a normal PVF to have LV abnormalities (18 of 22 versus 23 of 66, P=0.0005). However, a blunted PVF was not associated with clinically significant MR. CONCLUSIONS: A normal PVF does not rule out the absence of LV abnormalities but confirms the absence of significant MR. Systolic flow reversal is highly predictive of the presence of significant MR. A blunted PVF is more likely to be associated with LV abnormalities than with MR and has limited usefulness in the diagnosis of significant MR. PMID- 10375717 TI - Profile of cardiac tamponade in the medical emergency ward of a North Indian hospital. AB - OBJECTIVE: To determine the profile of patients presenting to the medical emergency ward with cardiac tamponade. DESIGN: Retrospective observational study. SETTING: Tertiary care hospital in North India. PATIENTS: Thirty patients (19 men and 11 women) presenting to the medical emergency ward with cardiac tamponade from March 1, 1995 to March 31, 1997. MAIN RESULTS: The mean age was 36.5+/-7.6 years for the men and 34+/-12.4 years for the women. Breathlessness, fever, cough, chest pain and easy fatigability were present in 97%, 90%, 70%, 57% and 37% of patients, respectively. Etiologically, tuberculosis accounted for 60%, malignant disease for 33% and hypothyroidism for 7% of cases of cardiac tamponade. Echocardiographically guided pericardiocentesis was carried out in all patients without any complications. Six patients underwent catheter pericardial drainage and, of these, four required pericardiostomy. CONCLUSIONS: Tuberculosis ranked as the most common cause of cardiac tamponade in Northern India, followed by malignancy. Therapeutically, echocardiographically guided pericardiocentesis for cardiac tamponade is a safe and effective procedure. For those with recurrent pericardial effusions, catheter pericardial drainage is a safe option until the underlying cause can be treated or surgery planned. PMID- 10375718 TI - Effect of a single bolus of intracoronary basic fibroblast growth factor on perfusion in an ischemic porcine model. AB - Basic fibroblast growth factor (bFGF) has been shown to induce angiogenesis in various animal models, but the methods of administration used experimentally are not clinically feasible. The objective of this study was to determine whether a single intracoronary bolus injection of bFGF would improve coronary perfusion in a porcine ischemic model that mimics clinical chronic ischemia. A copper coil studded with gold was delivered into the proximal right coronary artery of juvenile Yorkshire pigs and deployed by interventional techniques. After a four week interval for stenosis maturation, bFGF (100 micrograms) was administered by bolus injection into the left coronary artery in five animals, and vehicle alone was administered in four animals. Angiogenesis and change in right coronary perfusion area were assessed two weeks later by angiography, myocardial contrast echocardiography and immunohistochemistry. The right coronary perfusion area increased significantly after treatment in all but one of the animals that received bFGF but not in any of the controls. Intimal hyperplasia was not induced by bFGF. Capillary density determined histochemically was not different in the two groups. In conclusion, in a porcine ischemic model, bFGF administered by a single bolus intracoronary injection into the contralateral artery improved antegrade perfusion into the ischemic territory although without histological evidence of angiogenesis. This preliminary work merits further investigation. PMID- 10375719 TI - Alterations in protein kinase A and protein kinase C levels in heart failure due to genetic cardiomyopathy. AB - BACKGROUND: It is becoming evident that both cardiac and skeletal muscles are affected in congestive heart failure. Although protein kinases are known to regulate cardiac function, very little is known about their status in cardiac and skeletal muscles during the development of congestive heart failure. OBJECTIVE: To determine changes in the activities and protein levels of protein kinase A (PKA) and protein kinase C (PKC) in cardiac and skeletal muscles in congestive heart failure due to genetic cardiomyopathy on the basis that PKA and PKC are crucial for protein phosphorylation. ANIMALS AND METHODS: Genetically cardiomyopathic UM-X7.1 hamsters (250 to 300 days old) and age-matched Syrian hamsters were used in this study. PKA and PKC activities were assayed by measuring 32P from [gamma-32P]ATP incorporated into synthetic substrates. Relative protein contents of these protein kinases were obtained by using immunoblot analysis in control and failing hamster hearts and skeletal muscles. RESULTS: PKC activity was significantly increased in the failing hearts compared with control preparations. The relative protein contents of cytosolic PKC-alpha and -epsilon , and of particulate PKC-epsilon isozymes were significantly increased in failing hearts. PKC activity was also markedly increased in cardiomyopathic skeletal muscle. Furthermore, PKA activity and protein level in both cardiac and skeletal muscles were significantly increased in the failing heart group compared with control values. CONCLUSIONS: Increased PKC activity in heart failure may be due to changes in PKC-alpha and -epsilon isozymes in cardiomyopathic hearts. Alterations of PKA and PKC in congestive heart failure were not limited to the heart because similar changes in enzyme activities were evident in skeletal muscle. PMID- 10375720 TI - Profile: robert S fraser PMID- 10375721 TI - A crusty cause of prosthetic valve endocarditis. AB - A 73-year-old man with two previous mitral valve replacements presented with prosthetic valve infective endocarditis. Ten days before hospitalization he had undergone minimally invasive cutaneous surgery for crusty lesions but had not received antibiotic prophylaxis. The current literature regarding the role of antibiotic prophylaxis in dermatological procedures is discussed along with the issues surrounding this patient. PMID- 10375722 TI - Dilated cardiomyopathy as a first sign of nutritional vitamin D deficiency rickets in infancy. AB - A five-month-old boy presented with severe dilated cardiomyopathy, requiring intravenous inotropes as part of the initial management. He was found to have hypocalcemia due to vitamin D deficiency rickets. His cardiac function recovered completely after six months of vitamin D supplementation. PMID- 10375723 TI - Reversible biventricular dysfunction secondary to ischemia in a patient with acute airway obstruction: a case report and review of the literature on reversible causes of acute ventricular dysfunction. AB - Reversible causes of acute myocardial dysfunction are important for clinicians to recognize. Reversible biventricular dysfunction secondary to myocardial ischemia is presented in a patient with acute upper airway obstruction. The differential diagnosis of reversible acute myocardial dysfunction is reviewed. PMID- 10375724 TI - The second annual Dr robert E beamish award PMID- 10375725 TI - Dual effect of cobra cardiotoxin on vascular smooth muscle and endothelium. AB - AIM: To assess the cytotoxic effects of cobra cardiotoxin (CTX) on rat aorta. METHODS: Measure of contractility of aortic rings with or without endothelium. RESULTS: In endothelium-intact rings, CTX 10 mumol.L-1 induced a transient relaxation followed by a sustained contraction. Removal of the endothelium or pre incubation of the rings with NO synthase inhibitor NG-nitro-L-arginine methyl ester (L-NAME) abolished the transient relaxation but did not affect the magnitude of the contractile response induced by CTX. CTX itself induced contraction of vascular smooth muscle but also reduced contractions induced by phenylephrine (PhE) or KCl stimulation in a concentration-dependent manner. Contraction induced by CTX was dependent on the external Ca2+ concentration. Maximal contractile response to CTX was obtained in medium containing Ca2+ 1 mmol.L-1. This response decreased with higher Ca2+ concentration and disappeared when Ca2+ 7 mmol.L-1, organic and inorganic calcium channel blockers were present in the external solution before CTX addition. In preparations with the endothelium intact and incubated with CTX, relaxation by acetylcholine (ACh) stimulation of the tension induced by PhE was decreased. Endothelium-dependent relaxation to ACh was preserved when Ca2+ 5 mmol.L-1 was added to the medium prior to CTX. CONCLUSION: CTX first triggers the release of NO from the endothelium which results in muscle relaxation, and then causes smooth muscle contraction, Ca2+ and Ca2+ channel blockers prevented the effect of CTX. PMID- 10375726 TI - Hemolytic activity of copper sulfate as influenced by epinephrine and chelating thiols. AB - AIM: To study the effects of epinephrine, homocysteine, and other complexing agents on the cytotoxicity of copper sulfate. METHODS: In vitro suspensions of human red cells incubated with cupric sulfate were used, and hemolysis was determined by extracellular hemoglobin. RESULTS: The hemolytic activity of CuSO4 (0.3 mmol.L-1) was enhanced by the presence of epinephrine and to a lesser extent by homocysteine, whereas D-penicillamine, succimer, and mercaptodextran reduced the copper-induced hemolysis. The latter 3 chelating thiols also reduced the copper-epinephrine-induced hemolysis. The plasma protein ceruloplasmin reduced markedly the copper-epinephrine-induced hemolysis, even upon concentrations < 20% of that of copper. Chromic chloride, as well, acted anti-hemolytically. CONCLUSION: The latter protectors may interact with the production or activity of toxic oxygen, while classical copper chelators sequester cupric ions from interaction with epinephrine or homocysteine. PMID- 10375727 TI - Pyruvate-based peritoneal dialysate preserves neutrophilic oxygen consumption. AB - AIM: To investigate effects of pyruvate- or lactate-based peritoneal dialysis solutions (P-PDS or L-PDS) on neutrophilic oxygen consumption and the role of the extracellular pH (pHe) in cells' oxygen uptake. METHODS: Human neutrophils were incubated in P-PDS or L-PDS containing pyruvate or lactate 35-38 mmol.L-1 at various pHe, respectively. Oxygen consumption rates by opsonized zymosan (OZ) stimulated cells were measured polarographically, using a Clark-type oxygen electrode. RESULTS: L-PDS at an initial pH 5.2 dramatically inhibited the rate of oxygen consumption (2.2 nmol.min-1/10(6) cells) by neutrophils, while the equally acidic P-PDS markedly improved the rate (6.4 nmol.min-1/10(6) cells) (P < 0.01). However, P-PDS at pHe 5.2 severely impaired the rate by cells, the same as pHe 5.2 L-PDS. CONCLUSION: P-PDS preserved an oxygen consumption rate by OZ stimulated human neutrophils, but in an acidi milieu it comparably deteriorated the ability of cells to consume oxygen, indicating that the pHe of PDS plays an essential role in cellular oxidative metabolism. The superior biocompatibility of an acidic P-PDS was associated with its lower buffering capacity. PMID- 10375728 TI - Contractile effect of 6 beta-acetoxy nortropane on human and guinea pig airways. AB - AIM: To study the effects of 6 beta-acetoxy nortropane (6 beta-AN) on the isolated human bronchus and guinea pig trachea. METHODS: The contractile effect of 6 beta-AN was studied with 4 different muscarinic receptor antagonists on airway strips and inositol phosphates (IP) accumulation in human bronchi was determined by HPLC with radioactivity flow detector. RESULTS: (1) The maximal contractile effect of 6 beta-AN was lower than that of acetylcholine (ACh) on the human bronchus and equal to that of ACh on the guinea pig trachea. 6 beta-AN was more potent than ACh on both preparations (68 and 245 times, respectively). (2) The contractile effect of 6 beta-AN was inhibited by atropine (1 -100 nmol.L-1) or para-fluoro-hexahydro-sila-difenidol (0.01-1 mumol.L-1), but not by methoctramine (Met, 0.3-3 mumol.L-1) or pirenzepine (0.01-0.1 mumol.L-1), and was not enhanced by tacrine (0.1-10 mumol.L-1) or by epithelium removal. (3) The 6 beta-AN induced-contraction was accompanied by an increase of IP levels in isolated human bronchial tissues. (4) 6 beta-AN had an inhibitory effect on isoprenaline (Iso)-induced relaxation, which was abolished or reduced by Met 0.3 mumol.L-1. CONCLUSION: 6 beta-AN exerts a potent contractile effect involving muscarinic M3 receptor stimulation on airway smooth muscle. Muscarinic M2 receptor stimulation is furthermore partially involved in the antagonism by 6 beta-AN on the Iso-induced relaxation of the guinea pig trachea. PMID- 10375729 TI - Human mu-opioid receptor overexpressed in baculovirus system and its pharmacological characterizations. AB - AIM: To overexpress human mu-opioid receptor (muOR) with characteristics similar to those of mammalian origin. METHODS: Human muOR with a tag of 6 consecutive histidines at its carboxyl terminus was expressed in recombinant baculovirus infected Sf9 insect cells. Then the pharmacological characterizations of the product were studied by receptor binding assay and cAMP assay. RESULTS: The maximal binding capacity for the [3H]diprenorphine and [3H]ohmefentanyl (Ohm) were 9.1 +/- 0.7 and 6.52 +/- 0.23 nmol/g protein, respectively. The [3H]diprenorphine or [3H] Ohm binding to the receptor expressed in Sf9 cells was strongly inhibited by alpha-selective agonists [D-Ala2, N-methyl-Phe4, glyol5] enkephalin (DAGO), Ohm, and morphine, but neither by the delta-selective agonist [D-Pen2, D-Pen5] enkephalin (DPDPE) nor by the kappa-selective agonist ?trans-(+/ )-3, 4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl) cyclohexyl]? benzacetamide (U50488). NaCl 100 mmol.L-1 and guanosine triphosphate (GTP) 50 mumol.L-1 could reduce mu agonists Ohm and etorphine affinity binding to the expressed muOR. DAGO and Ohm effectively inhibited forskolin-stimulated cAMP accumulation. This agonist-dependent effect was blocked by opioid antagonist naloxone. CONCLUSION: The overexpression of human muOR with a tag of six consecutive histidines at its carboxyl terminus in Sf9 insect cells retained the characteristics of wild-type human muOR. PMID- 10375730 TI - Neuroprotective action of dextromethorphan in rat photochemically-induced focal cerebral ischemia. AB - AIM: To study the effects of dextromethorphan (Dex) on photochemically-induced focal cerebral ischemia in rats. METHODS: Anesthetized rats undergone 10-min light irradiation on exposed skull after rose bengal injection were pretreated with saline and Dex at 3 doses (12.5, 25, and 50 mg.kg-1, i.p., 15 min before ischemia). The alteration of volume of lesioned cortical region, regional cerebral blood flow (CBF), bcl-2 and bax expression at penumbra area were studied. RESULTS: Dex dose-dependently decreased the infarcted volume (17%, 26%, and 50% reduction, respectively). Pretreatment with Dex at a dose of 50 mg.kg-1 improved the postischemic hypoperfusion compared with the control at 20 and 30 min after lesion (both 31% increase), and also upregulated the expression of anti apoptosis gene bcl-2. CONCLUSION: Dex protects against ischemic neuronal damage in this model and its effects on CBF and bcl-2 expression may contribute to its neuroprotective action. PMID- 10375731 TI - Isolation of rat sacral dorsal commissural neurons. AB - To isolate rat sacral dorsal commissural neurons (SDCN). METHODS: Using enzymatic and mechanical dissociation techniques to isolate the neurons and using nystatin perforated patch technique to evaluate their functional state. RESULTS: The isolated neurons exhibited good responses to excitatory and inhibitory amino acids. The responses of SDCN to N-methyl-D-aspartate were markedly potentiated by substance P and trans-1-aminocyclopentane-1,3-dicarboxylate, whereas GABA responses were significantly potentiated by diazepam, pregnenolone, and pentobarbital. CONCLUSION: This preparation provides a satisfactory model for exploring the mechanisms of the SDCN in nociception and antinociception. PMID- 10375732 TI - Analysis of multidrug effects by parameter method. AB - AIM: To set up a new analytic method for multidrug effects. METHODS: Based on the principles of the target site kinetics and the equieffective test, a new mathematical model was set as Q = (Eo-Ee)/magnitude of Ee x W-sx x T (-1 < Q < 1 addition, Q < or = -1 antagonism, Q > or = 1 synergism) where Eo = a fitted value of the observed effect of a combination, Ee = an expected value of combined effect, W = an equieffective criterion decided by a special field, sx = a common standard error of Eo and Ee, and T = a value of one-sided t0.05. All the calculations were completed with computer. Dose-effect data from different types of experiments were fitted by the new model and the results were compared with those of other methods. RESULTS: This parameter method dealt with different types of data well fitted with the Hill equation, and was not limited to analyze receptor interaction of drugs, or the number of combined drugs. A series of Q values was obtained from all levels of dose-effect for a systematic analysis. The analysis took the criterion of a special field and laboratory error into account. CONCLUSION: This parameter method can effectively analyze the multidrug effects. PMID- 10375733 TI - Effects of taurine on L-type voltage-dependent Ca2+ channel in rat cardiomyocytes infected with coxsackievirus B3. AB - AIM: To study the effects of taurine on L-type voltage-dependent Ca2+ channel (VDCC) in adult rat cardiomyocytes infected with coxsackievirus B3 (CVB3). METHODS: Whole-cell Ca2+ current of L-type VDCC was obtained by patch-clamp techniques. RESULTS: The density of L-type Ca2+ current was 4.1 +/- 0.8 pA/pF in normal cardiomyocytes, but increased to 4.9 +/- 1.4 pA/pF with CVB3 infection. At 16 mmol.L-1, taurine decreased the density to 3.5 +/- 0.5 pA/pF in normal cardiomyocytes, and to 3.8 +/- 0.8 pA/pF in CVB3-infected cardiomyocytes. In addition, CVB3 shifted the membrane potential depolarizing to peak current (Vp) from 8 +/- 8 mV to 5 +/- 3 mV which could also be reverted to 8 +/- 4 mV by taurine. CONCLUSION: Taurine inhibited the increase of Ca2+ inflow through L-type VDCC and normalized the decreased Vp induced by CVB3 infection. The effect of taurine on L-type VDCC was the mechanism of taurine attenuating the intracellular Ca2+ accumulation and abnormal electric activities induced by CVB3 infection. PMID- 10375734 TI - Bradykinin B2 receptor antagonist icatibant reduces inhibitory effect of captopril on growth of cultured neonatal rat cardiomyocytes. AB - AIM: To study whether endogenous kinins are negative modulators in the growth of cardiomyocytes and their possible cellular and molecular mechanisms. METHODS: Cultured neonatal rat cardiomyocytes were used. Intracellular RNA and protein synthesis were measured by [3H]uridine incorporation and [3H]leucine incorporation, respectively. The expression level of proto-oncogene c-myc, c-fos mRNA was observed by Northern blotting. RESULTS: Exposure of cardiomyocytes to captopril (Cap, 100 mumol.L-1) for 48 h inhibited the rates of [3H]Urd and [3H]Leu incorporations by 25% and 26%, respectively, and for 2 h inhibited c-myc, c-fos mRNA expression by 75% and 55%, respectively. Treatment of angiotensin II (Ang II, 1 mumol.L-1) for 48 h significantly increased the rates of [3H]Urd and [3H]Leu incorporations and for 1 h induced c-myc, c-fos mRNA overexpression, which were reduced by pretreatment with Cap (100 mumol.L-1). Icatibant acetate (Hoe 140, a specific antagonist of bradykinin B2 receptor) 0.1-10 mumol.L-1 blocked the effects of Cap in a concentration-dependent manner. CONCLUSION: Endogenous kinins exhibited a negative modulatory effect on growth of cardiomyoctes via BK B2 receptor. PMID- 10375735 TI - Oxidized low-density lipoproteins induce apoptosis in vascular smooth muscle cells. AB - AIM: To examine whether oxidized low density lipoproteins (ox-LDL) could induce apoptosis in rabbit aortic smooth muscle cells (VSMC). METHODS: Low density lipoproteins (n-LDL) were isolated from healthy human plasma by gradient ultracentrifugation and oxidized by CuSO4 10 mumol.L-1. VSMC were exposed to ox LDL, n-LDL, or phosphate-buffer solution (PBS) as control. Morphological changes were observed under fluorescene microscope after Hoechst 33258 staining. Extracted DNA was electrophoresized on agarose gel. RESULTS: Incubation of VSMC with ox-LDL 300 mg.L-1, not n-LDL, for 24 h induced morphological apoptosis changes (chromatin condensation, nucleus fragmentation) and DNA fragmentation, which was furthered with the incubation time up to 48 h or at a concentration of 400 mg.L-1. Dextran sulfate, a scavenger receptor blocker and butylated hydroxytoluene (BHT), an antioxidant, exhibited no effect on DNA fragmentation. Lysophosphatidylcholine (LPC) at a concentration up to 125 mumol.L-1 (equivalent to ox-LDL 300 mg.L-1) did not elicit DNA fragmentation. CONCLUSION: Ox-LDL induced apoptosis in VSMC without involving oxygen free radicals and LPC. PMID- 10375736 TI - Effects of sex hormones on action potential and contraction of guinea pig papillary muscle. AB - AIM: To study the effects of sex hormones, estradiol (Est), progesterone (Pro) and testosterone (Tes) on the action potential (AP) and contraction of guinea pig papillary muscle. METHODS: Using conventional glass microelectrode and mechanical recording of myocardial contraction. RESULTS: Est slowed down the maximal rate of rise of phase 0 (Vmax) of AP at low concentration (1 mumol.L-1). At 10 mumol.L-1 and above, Est also prolonged AP duration (APD50 and APD90), effective refractory period (ERP) and decreased the maximal isometric tension (Pmax) and velocity of tension development (dT/dt) of contraction. Tes (100 mumol.L-1 - 1 mmol.L-1) prolonged APD90 and ERP with decreased Pmax and dT/dt. But Pro (1 mumol.L-1 - 1 mmol.L-1) had no effects on both AP and contraction. CONCLUSION: Est prolonged AP and depressed contraction of guinea pig papillary muscle. PMID- 10375737 TI - 8-(N,N-diethylamino)-n-octyl-3,4,5-trimethoxybenzoate reduced [Ca2+]i elevation induced by histamine, serotonin, and glutamate in cultured calf basilar artery smooth muscle cells. AB - AIM: To study the effects of 8-(N,N-diethylamino)-n-octyl-3,4,5- trimethoxybenzoate (TMB-8) on intracellular free calcium ([Ca2+]i) in cultured calf basilar artery smooth muscle cells. METHODS: [Ca2+]i was examined by a system of measurement of AR-CM-MIC, using Fura 2-AM as a fluorescent indicator. RESULTS: In the presence of extracellular Ca2+ 1.3 mmol.L-1, histamine (His), serotonin (5-HT), and sodium glutamate (Glu) markedly increased the [Ca2+]i which was attenuated by TMB-8. In Ca2+ free Hanks' solution containing egtazic acid 0.1 mmol.L-1, TMB-8 not only reduced the resting [Ca2+]i, but also inhibited the elevation of [Ca2+]i evoked by His and 5-HT. CONCLUSION: TMB-8 reduced the resting [Ca2+]i and attenuated His-, 5-HT-, and Glu-induced increases of [Ca2+]i in basilar artery smooth muscle cells. PMID- 10375738 TI - Inhibitory effect of trapidil on proliferation of cultured rat aortic smooth muscle cells induced by endothelin-1. AB - AIM: To study the effect of trapidil (Tra) on endothelin-1-induced proliferation of cultured rat aortic vascular smooth muscle cells (VSMC). METHODS: The [3H]TdR incorporation into DNA assay, the number of VSMC, and cell cycle distribution were measured. RESULTS: Pretreated with endothelin-1 100 nmol.L-1, cell number, [3H]TdR uptake, and cell mitogenic activity increased 134% +/- 23%, 210% +/- 70%, and 86% +/- 18%, respectively. This proliferation was inhibited by Tra 5, 50, 500 mumol.L-1. The inhibitory rates were 12%-48%, 35%-54% and 15%-47%, respectively. Tra did not influence the proliferation of VSMC without endothelin-1 pretreatment. CONCLUSION: Tra antagonized the proliferation of VSMC induced by endothelin-1. PMID- 10375739 TI - Cytokine and nitric oxide production by rat microglia stimulated with lipopolysaccharides in vitro. AB - AIM: To study the characterization of interleukin (IL)-1, IL-2, tumor necrosis factor-alpha (TNF-alpha), and nitric oxide (NO) production in microglia stimulated with lipopolysaccharides (LPS). METHODS: Primary cultured neonatal rat microglia were incubated with LPS (0-10 mg.L-1) for 0-72 h. The supernatants and lysates were collected. IL-1, IL-2, and TNF-alpha were assayed by mouse thymocyte proliferation, mouse spleen cell proliferation, and 1929 cytotoxity, respectively. NO was assayed by Griess reaction. RESULTS: Extracellular IL-1, TNF alpha, and NO production reached peak levels at LPS 1 mg.L-1. Intracellular IL-1 production reached its peak level at LPS 100 micrograms.L-1. Intracellular TNF alpha level was very low. IL-1, TNF-alpha, and NO activities were detected at 1, 4, and 8 h, after the cells were stimulated with LPS. IL-1 got to its peak value at 8 h, TNF-alpha, and NO reached the highest levels at 24 h. However, IL-2 activity was not detected after the microglia were stimulated with LPS 0-10 mg.L 1 during the incubation period. CONCLUSION: Rat microglia stimulated with LPS in vitro produced proinflammatory cytokines and NO. PMID- 10375740 TI - Promotion of ATP and S-140 to ribosome inactivation with camphorin, cinnamomin, and other RNA N-glycosidases. AB - AIM: To study the effect of ATP and extra-ribosomal factors (S-140) on type I and type II RNA N-glycosidases in inactivating ribosome. METHODS: The activity of ATP and S-140 was determined by characterization of R-fragment in gel. An improved two-step method of cell-free protein synthesis system was used to quantitate the requirements of S-140 in ribosome inactivation. RESULTS: IC50 ratios of camphorin, gamma-momorcharin, luffin S, luffin A, trichosanthin (type I); and ricin, ricin A-chain; cinnamonin, cinnamomin A-chain (type II) between the absence and presence of ATP and S-140 were 3108, 151, 51, 45, 15; and 47, 7, 26, 12, respectively. CONCLUSION: The ribosome-inactivating activity of type II ribosome-inactivating proteins, including intact protein and its A-chain, was promoted by ATP and S-140. Camphorin showed a significant difference from cinnamomin in need of ATP and S-140 for such promoting. PMID- 10375741 TI - Antisense expression of protein kinase C alpha improved sensitivity to anticancer drugs in human lung cancer LTEPa-2 cells. AB - AIM: To study the role of protein kinase C alpha (PKC alpha) in sensitivity to some clinical anticancer drugs in human lung cancer LTEPa-2 cells. METHODS: Human lung cancer cell model expressing antisense PKC alpha was established and characterized by gene transfection and immunoblotting. Northern blotting was used to analyze the expression of multiple drug resistance (mdr-1) gene and antisense PKC alpha mRNA. IC50 for some anticancer drugs in cultured cells were measured. RESULTS: Expression of antisense PKC alpha mRNA inhibited mdr-1 gene expression in lung cancer cells and improved sensitivity to anticancer drugs (harringtonine, carboplatin, bleomycin A5, vincristine and doxorubicin) in lung cancer cells. IC50 for harringtonine, carboplatin, bleomycin A5, vincristine, and doxorubicin was decreased by 46.4%, 42.1%, 79%, 69.9%, and 61.6% respectively. CONCLUSION: PKC alpha plays an important regulation role of mdr-1 gene expression and drug sensitivity in human lung cancer cells. PMID- 10375742 TI - Artemisinin blocks activating and slowly activating K+ current in guinea pig ventricular myocytes. AB - AIM: To study the effect of artemisinin (Art) on outward rectifier potassium current in ventricular myocytes. METHODS: In isolated guinea pig ventricular myocytes, the effects of Art on the two components of delayed outward rectifier K+ current (IK), the rapidly activating inward K+ current (IKr), and the slowly rectifying outward K+ current (IKs) were observed by the whole cell patch-clamp technique. RESULTS: Art decreased IK in a concentration-dependent manner. The IKstep and IKtail were reduced from 387 +/- 46 pA to 240 +/- 48 pA and from 299 +/- 30 pA to 130 +/- 38 pA, respectively at holding potential of +40 mV by Art 50 mumol.L-1. The envelope of tail analysis suggested that both IKr and IKs were inhibited. CONCLUSION: Art blocked the two components of delayed outward rectifier K+ current (IKr and IKs) in guinea pig ventricular cells. PMID- 10375743 TI - Menadione reduced doxorubicin resistance in Ehrlich ascites carcinoma cells in vitro. AB - AIM: To study the effect of menadione (Men) reducing doxorubicin (Dox) resistance in Ehrlich ascites carcinoma (EAC) cells resistant to Dox (EAC/Dox cells). METHODS: Glutathione (GSH) content and membrane fluidity were measured by fluorometric assay and fluorescence depolarization assay, respectively. Glutathione S-transferase (GST) activity was measured with 1-chloro-2,4 dinitrobenzene as the substrate. Cell viability was determined by 3-(4, 5 dimethylthiazol)-2, 5-diphenyltetrazolium bromide assay. RESULTS: GSH content, GST activity, and membrane fluidity in EAC/Dox cells were higher than those in EAC cells (P < 0.01). The IC50 (95% confidence limits) for Dox on EAC/Dox cell was 22.3 (15.8-28.8) mg.L-1. Relative resistance of Dox in EAC/Dox cells was 42 fold. Pretreatment of EAC/Dox cells with Men 5 or 10 mg.L-1 decreased intracellular GSH content (P < 0.01). Men 1 mg.L-1 had no obvious effect on GSH content in EAC/Dox cells (P > 0.05), but decreased the elevated membrane fluidity efficiently (P < 0.05). Men had no obvious effect on GST activity in EAC/Dox cells (P > 0.05). IC50 of Dox was reduced to 9.6 (7.8-11.3), 6.0 (2.8-9.2), or 5.3 (3.9-6.7) mg.L-1 in EAC/Dox cells pretreated with Men 1, 5, or 10 mg.L-1. CONCLUSION: Men reduced Dox resistance effectively due in part to its depletion of GSH content in EAC/Dox cells. PMID- 10375744 TI - Negative correlation between cyclophilin mRNA level in leukocytes of renal allograft recipients and ciclosporin concentration in whole blood. AB - AIM: To study mRNA level of cyclophilin in the white blood cells (WBC) of the renal allograft recipients (RAR) and its correlation with ciclosporin concentration in whole blood. METHODS: The cyclophilin mRNA levels and beta-actin as controls in the WBC of 47 RAR were measured by quantitative reverse transcription polymerase chain reaction. The blood ciclosporin assay utilized monoclonal antibody fluorescence polarization immunoassay. RESULTS: With the increase of ciclosporin concentration in whole blood (from 62 micrograms.L-1 to 678 micrograms.L-1), relative cyclophilin mRNA level in the WBC of RAR decreased nonlinearly (from 1.1 to 0.03, r = 0.8195). CONCLUSION: There was a negative correlation between the mRNA level of cyclophilin in the WBC of RAR and the ciclosporin concentration in whole blood. PMID- 10375745 TI - Effect of artemether on glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, and pyruvate kinase of Schistosoma japonicum harbored in mice. AB - AIM: To study the effect of artemether (Art) on glyceraldehyde-phosphate dehydrogenase (GAPDH), phosphoglycerate kinase (PGK), and pyruvate kinase (PK) of S japanicum. METHODS: Mice infected with schistosome cercariae for 32-38 d were treated ig with Art 100-300 mg.kg-1 and killed 24-72 h after medication for collection of schistosomes. The activities of GAPDH, PGK, and PK of the worms were determined by measuring the formation of NADH or consumption of NAD. The lactate content of the worms was also measured. RESULTS: After the infected mice were treated ig with Art 300 mg.kg-1 for 24 h, the inhibition rates of GAPDH were 13% (Male) and 21% (Female), and 48 h later the inhibition rates of the enzyme were 6% (Male) and 28% (Female). When Art 300 mg.kg-1 was given to infected mice for 24 h and 48 h, the inhibition rates of PGK were 60% (Male) and 48% (Female) as well as 75% (Male) and 62% (Female), respectively. Similar results were seen in PK activity. At 72 h after treatment the reduction rate of lactate content in Female worm was 72%, while that of Male was 48%. CONCLUSION: In the glycolytic pathway of both Male and Female schistosomes, PGK and PK activities were inhibited by Art. The GAPDH activity of Female worms was also susceptible to Art, While that of Male worms showed only temporary inhibition after treatment with Art. The Art reduced lactate content more in Female than in Male worms. PMID- 10375746 TI - [Antimutagenic effects of beta-carotene from Dunaliella salina]. AB - AIM: To study the genotoxic and antimutagenic efficts of beta-carotene from Dunaliella salina (beta-CDS). METHODS: The in vitro micronucleus and chromosomal aberration tests in hunman lymphocytes were adopted. The effect of beta-CDS on mutagensis induced by gamma-rays and mitomycin (Mit) was studied. RESULTS: beta CDS (1-30 mg.L-1) had no genotoxicity, but inhibited spontaneous and gamma-ray induced micronucleus fromation and Mit-induced chromosomal aberrations in human lymphocytes in vitro. The genotoxic action of synthetic beta-carotene (S beta C) was suppressed, the antimutagenic effects were heightened when S beta C and beta carotene oil (beta CO, one of the beta-CDS compositions) were mixed in proportion as 80:27.5. CONCLUSION: beta-CDS is an antimutagenic agent. PMID- 10375747 TI - Immunomodulating effects of methionine enkephalin. AB - Methionine enkephalin (Met-Enk), the endogenous neuropeptide, is suggested to be involved in the regulatory loop between immune and neuroendocrine systems. Our studies showed that Met-Enk over a wide range of concentrations increased interleukin-1 (IL-1) production from mouse peritoneal macrophages induced by lipopolysaccharides (LPS) and naloxone did not block the enhancing effect. Met Enk promoted the proliferation of mouse splenocyte and the production of IL-2 and IL-6 in a dose-dependent manner. The up-regulating effects of IL-2 and IL-6 not only augmented their mRNA transcription but also increased their stability. Thus Met-Enk appears to be an important immunomodulatory signaling molecule to exert regulatory actions concerned with the expressing of pre-inflammatory cytokines. PMID- 10375748 TI - Pharmacokinetics of 5-fluorouracil and its penetration into pancreatic juice in dogs. AB - AIM: To study the pharmacokinetic behavior of 5-fluorouracil (5-FU) in pancreatic juice in dogs and its correlation with 5-FU in plasma, and to evaluate its penetration characteristics. METHODS: After placing a pancreatico-drainage tube, 8 dogs were injected 5-FU 250 mg i.v. Blood and pancreatic samples were collected and the 5-FU concentrations were determined by HPLC. The pharmacokinetic parameters were obtained with statistical analysis. RESULTS: The mean slopes of the terminal phase (K10) in plasma and pancreatic juice were 9.4 h-1 and 10.2 h 1, respectively (P > 0.05). The pharmacokinetic behaviors of 5-FU in plasma and pancreatic juice fitted a nonlinear model. Its penetration index was 3.39 +/- 2.84. The penetration of 5-FU from blood to pancreatic juice was relatively rapid, demonstrating a consistently higher concentration in pancreatic juice than in plasma. CONCLUSIONS: The elimination phase of 5-FU in plasma was similar to that in pancreatic juice, indicating that they were in the same kinetic compartment. PMID- 10375750 TI - Involvement of a putative G-protein-coupled receptor and a branching pathway in argipressin (4-8) signal transduction in rat hippocampus. AB - AIM: To study the signal transduction pathway induced by argipressin (4-8) (AVP4 8) in rat hippocampus. METHODS: Rat hippocampi were sectioned transversely at 300 microns with a tissue chopper and transferred to fresh incubation solution circulated with a humidified gas mixture of 95% O2 + 5% CO2 at 36 +/- 0.5 degrees C. After incubation with various drugs, MAP kinase (MAPK) activity and Ca2+/calmodulin-dependent protein kinase II (CaMKII) autophosphorylation were measured. RESULTS: The main findings are: (1) The AVP4-8-stimulated MAPK activity and the CaMKII autophosphorylation were blocked by ZDC(C)PR, an antagonist of AVP4-8, and also completely inhibited by pertussis toxin, a selective inhibitor of the G-protein-coupled receptor (GPCR). But, AVP-induced MAPK activation was not sensitive to ZDC(C)PR or PTX. (2) Polymyxin B (PMB), an inhibitor of protein kinase C (PKC), markedly suppressed the peptide-activation of MAPK, but did not affect CaMKII autophosphorylation. Phorbol myristate acetate (TPA), an activator of PKC, elicited an increase of MAPK activity, but did not further influence the level of AVP4-8-enhanced MAPK activity; Nevertheless, the extent of CaMKII activation was attenuated by TPA. (3) The enhancement of MAPK activity was not reduced by KN-62, a specific inhibitor of CaMKII. (4) AVP4-8 did not show any influence on cAMP production. CONCLUSION: AVP4-8 stimulated signal transduction via a GPCR and a branching pathway in rat hippocampus. PMID- 10375749 TI - Antagonistic effect of orphanin FQ on opioid analgesia in rat. AB - AIM: To study the effect of orphanin FQ (OFQ), a newly discovered heptadecapeptide, on nociception and opioid analgesia. METHODS: The intracerebroventricular (i.c.v.) and intrathecal (i.t.h.) injections were used to give the drugs. The tail-flick model of rats were used to test the pain threshold. RESULTS: OFQ (i.c.v. or i.t.h.) 0.1 microgram had no effect on nociception but 0.5-10 micrograms induces hyper-reaction of rat to noxious electric stimulus; the decapeptide (OFQ1-10 i.c.v.), a fragment of the OFQ, did not affect the pain reaction of rats. Fentanyl (1 microgram, i.c.v. or i.t.h.), a selective mu-receptor agonist, DSLET (5 micrograms, i.c.v. or i.t.h.), a selective delta-receptor agonist, or U50488H (1 microgram, i.t.h.), a kappa receptor agonist, induced an increase in pain threshold, when OFQ (0.1 or 1 microgram) was added together with one of them (except for the ith injection of DSLET), the increase of pain threshold was reduced obviously. CONCLUSION: OFQ induces hyperalgesia and antagonizes opioid analgesia mediated by mu- and delta receptors in the brain and by mu- and kappa- but not delta-receptors in the spinal cord of rats. PMID- 10375751 TI - Involvement of medullary tail-flick related neurons in descending facilitation evoked by chemical stimulation of rat lateral habenular nucleus. AB - AIM: To study effects of sodium L-glutamate microinjection into lateral habenular nucleus (LHN) of rats on the firing of medullary tail-flick related neurons and tail-flick reflex (TF). METHODS: Using synchronous recording of unitary neuronal discharges and TF induced by noxious heat. RESULTS: Chemical stimulation of LHN induced an excitement of the on-cell spontaneous activity, an inhibition of the off-cell spontaneous activity with an enhancement of their TF related responses. The spontaneous firing rate of on-cells increased from 5.8 +/- 2.2 Hz to 10.9 +/- 3.4 Hz while the spontaneous firing rate of off-cells decreased from 11.8 +/- 2.2 Hz to 6.1 +/- 2.2 Hz. Meanwhile the TFL was shortened from 4.04 +/- 0.17 s to 2.97 +/- 0.13 s. CONCLUSION: The chemical stimulation of LHN produced a facilitating action on nociceptive spinal defensive reflex. This effect is brought out by the cooperation of on- and off-cells. PMID- 10375753 TI - Anticholinesterase effects of huperzine A, E2020, and tacrine in rats. AB - AIM: To compare the anticholinesterase effects of huperzine A (Hup A), E2020, and tacrine in rats. METHODS: Spectrophotometry was used to determine AChE activity in brain and BuChE activity in serum. RESULTS: Following intragastric gavage, Hup A, E2020, and tacrine all produced dose-dependent inhibitions of brain AChE. Oral Hup A exhibited a higher inhibition than E2020 and tacrine. Tacrine was more effective in inhibiting serum BuChE correlated with severe peripheral adverse effects. The BuChE activity was less affected by Hup A and E2020. After a single oral dose of Hup A, a relatively steady state of AChE inhibition produced, which was longer than that after E2020 and tacrine. No change in the cholinesterase inhibition was seen for the 3 drugs following repeated i.g. medications. CONCLUSION: Hup A i.g. exhibited a higher efficacy, a longer duration of action, and a more selective inhibition on AChE than E2020 and tacrine. PMID- 10375752 TI - Effects of N-methyl berbamine on delayed outward potassium current in isolated rat hepatocytes. AB - AIM: To study the effects of N-methyl berbamine (NMB) on the delayed outward potassium currents (Ik) in isolated rat hepatocytes. METHODS: With patch-clamp techniques and whole-cell recording method, holding potential -50 mV, command potential +30 to +140 mV, duration 900 ms. RESULTS: NMB reduced Ik in a concentration-dependent manner. When the concentrations of NMB were 20, 50, 400 nmol.L-1 and 50 mumol.L-1, the amplitude values of Ik were decreased to 3.6 +/- 0.4 (P > 0.05), 2.1 +/- 1.6 (P > 0.05), 3.7 +/- 1.6 (P < 0.05), 2.3 +/- 1.3 nA (P < 0.01) from 4.4 +/- 1.0 (n = 4), 2.5 +/- 1.8 (n = 4), 5.8 +/- 2.1 (n = 5), 4.6 +/- 1.3 (n = 6) nA of control, respectively. The inhibitory rates were 10%, 15%, 37%, and 51%, respectively. CONCLUSION: NMB was a K+ channel inhibitor. PMID- 10375754 TI - Pinacidil suppression on 5-HT3 receptor-mediated contraction of guinea pig ileum in vitro. AB - AIM: To study the effects of the K+ channel opener pinacidil on 5-HT3 receptor mediated contractions of the isolated guinea pig ileum (GPI) longitudinal muscle myenteric plexus strip preparations. METHODS: GPI contractions were recorded with a chart recorder through isometric transducers. The effect of pinacidil on binding properties of 5-HT3 receptors was assessed using [3H]GR65630 binding assay in membrane preparations of rat entorhinal cortex. RESULTS: (1) A selective 5-HT3 receptor agonist 2-methyl-5-HT 0.1-300 mumol.L-1 and 5-HT 0.001-50 mumol.L 1 elicited GPI contractile responses in concentration-dependent manners, the EC50 values (and 95% confidence limits) for 2-methyl-5-HT and 5-HT were 10.0 (8.9 11.2) mumol.L-1 and 1.6 (1.3-1.9) mumol.L-1, respectively. Selective 5-HT3 receptor antagonist tropisetron 0.1 mumol.L-1 competitively inhibited the responses to 2-methyl-5-HT and 5-HT. (2) Pinacidil 0.5-5 mumol.L-1 inhibited 5 HT3 receptor-mediated contractions. (3) Pinacidil 1 mumol.L-1 enhanced the inhibitory effects of tropisetron 0.1 mumol.L-1 or another selective 5-HT3 receptor antagonist benesetron 1 mumol.L-1 on 5-HT-induced GPI contractile responses. (4) Pinacidil 1-5 mumol.L-1 did not affect GPI contractile responses evoked by a selective M-ACh receptor agonist carbachol 1 mumol.L-1. (5) Pinacidil 1-5 mumol.L-1 had no effect on binding properties of 5-HT3 receptors with selective 5-HT3 receptor radioligand [3H]GR65630 in the entorhinal cortex of rat brain. CONCLUSION: The inhibition by pinacidil of 5-HT3 receptor-mediated GPI contractile responses may be mediated through activation of ATP-sensitive K+ channels located in prejunctional myenteric neurons. PMID- 10375755 TI - Effects of long-term application of dopamine HCl on dopamine agonist-induced cAMP production in rat renal cortex. AB - AIM: To study the effects of long-term application of dopamine HCl (DA) on the functional changes of dopamine receptor subtypes coupled to adenyl cyclase in rat renal cortex. METHODS: cAMP levels were measured by radioimmunoassay as an index of dopamine receptor function. RESULTS: Injection of DA (30 mg.kg-1.d-1, i.p. 30 d) reduced the fenoldopam (Fen) (100 mumol.L-1)-induced increments of cAMP production from the control group of +1.26 +/- 0.04 to the DA-treated group of +0.63 +/- 0.22 nmol.min-1/g tissue and the propyl-butyl-dopamine (PBDA) (100 mumol.L-1)-induced decrements of cAMP production in the presence of Sch-23390 (Sch) from the control group of -0.38 +/- 0.18 to the DA-treated group of -0.11 +/- 0.08 nmol.min-1/g tissue with, however, comparable percentile changes for the 2 groups. Sch blocked both Fen- and PBDA-induced increase in cAMP production, while domperidone (Dom) blocked the decreasing effects of PBDA on cAMP accumulation in the presence of Sch. CONCLUSION: Long-term application of DA produced a marked "down regulation" of both DA1 and DA2 receptors in rat renal cortex with, however, the responsiveness of the remaining receptors unchanged. PMID- 10375756 TI - Pharmacokinetics of flutamide and its metabolite 2-hydroxyflutamide in normal and hepatic injury rats. AB - AIM: To develop a new HPLC assay to study the pharmacokinetics of flutamide (Flu) and its active metabolite 2-hydroxyflutamide (HF) in rats. METHODS: Normal or hepatic injury rats were given i.g. Flu 50 mg.kg-1. Reverse phase HPLC was developed with a mu-Bondapak C 18 column. Internal standard was methyltestosterone. The mobile phase was a mixture of methanol:acetonitrile:water:diethyl ether = 40:20:35:1 (vol). Absorbance was measured at lambda 234 nm. RESULTS: The pharmacokinetic parameters of Flu were as follows: in normal rats, K = 0.62 +/- 0.16 h-1, Cl = 6.0 +/- 1.0 L.kg-1.h-1, AUC = 8.6 +/- 1.3 mg.L-1.h, Cmax = 2.4 +/- 0.7 mg.L-1; in hepatic injury rats, K = 0.16 +/- 0.03 h-1, Cl = 0.63 +/- 0.29 L.kg-1.h-1, AUC = 100 +/- 44 mg.L-1.h, Cmax = 6.7 +/- 2.8 mg.L-1. The pharmacokinetic parameters of HF were as follows: in normal rats, K(m) = 0.07 +/- 0.01 h-1, AUC = 219 +/- 22 mg.L-1.h, Cmax = 8.6 +/- 0.6 mg.L-1; in hepatic injury rats, K(m) = 0.05 +/- 0.01 h-1, AUC = 170 +/- 42 mg.L-1.h, Cmax = 3.8 +/- 0.8 mg.L-1. There were significant differences between the parameters of normal and hepatic injury rats (P < 0.01) except AUC of HF (P > 0.05). CONCLUSION: This HPLC assay was sensitive and precise, and the elimination of Flu and HF was inhibited significantly due to hepatic injury. PMID- 10375757 TI - Use of caffeine as a probe for rapid determination of cytochrome P-450 CYP1A2 activity in humans. AB - AIM: To develop a rapid HPLC method for the determination of cytochrome P-450 CYP1A2 activity. METHODS: A 300-microL plasma was prepared by extraction with 5 mL chloroform/isopropanol (9:1), and beta-hydroxyletheophylline was added as internal standard (IS). Samples were separated on an ODS column by a gradient elution system, of which mobile phase consisted of 0.05% acetic acid, acetonitrile, and methanol. The compounds of interest were monitored at 282 nm by UV detector. RESULTS: No potential interfering peaks were found. Paraxanthine (17X), IS and caffeine (137X) were rapidly eluted with baseline resolution, and their retention time was less than 13 min. The detection limits of both 17X and 137X were 0.1 mumol.L-1. Linear relations ranged over 1-100 mumol.L-1 and 1-200 mumol.L-1 with correlation coefficient of 0.9999 and 0.9987, respectively, for 17X and 137X. The coefficients of variation were within 6% for 17X, and 10% for 137X. The average recoveries for both compounds were ranged from 96% to 108%. CONCLUSION: This method is sensitive and rapid, and can be used for population studies of CYP1A2. PMID- 10375758 TI - Effects of tanshinone II-A sulfonate on adhesion molecule expression of endothelial cells and platelets in vitro. AB - AIM: To study the action of tanshinone II-A sulfonate (Tan) on adhesion molecule expression by cultured endothelial cells and platelets. METHODS: Tumor necrosis factor alpha (TNF-alpha)-induced ICAM-1 expression on the cell surface and endothelial adhesivity toward HL-60 cells were studied using human umbilical vein endothelial cells (HUVEC). Thrombin-induced expression of platelet P-selectin was studied using human blood platelets. Adhesion molecule expression on the cell surface was measured by flow cytometry. The number of HL-60 cells adhering to the HUVEC monolayer was determined by liquid scintillation spectroscopy. RESULTS: Pretreatment of HUVEC with TNF-alpha significantly enhanced ICAM-1 expression and increased HL-60 cells adhesion to HUVEC from 4.6% +/- 0.7% to 30% +/- 6%. Tan (25 200 mumol.L-1) inhibited the effects of TNF-alpha in a concentration-dependent manner. Tan also inhibited the increase of P-selectin expression of thrombin activated platelets in a concentration-dependent manner. CONCLUSION: Tan inhibited expression of adhesion molecules (ICAM-1, P-selectin) in HUVEC and in human blood platelets. PMID- 10375759 TI - Pharmacokinetics of recombinant human granulocyte macrophage colony-stimulating factor in Macaca mulatta. AB - AIM: To examine the pharmacokinetics of i.v. and s.c. recombinant human granulocyte macrophage colony-stimulating factor (rhGM-CSF) in Macaca mulatta. METHODS: Plasma levels of rhGM-CSF were detected with sandwich enzyme-linked immunosorbent assay. RESULTS: Plasma concentration-time curves after i.v. rhGM CSF in monkeys were best fitted with 3-compartment model. The 1st, 2nd, and 3rd phase T1/2 were 0.05-0.07, 0.14-0.58, and 1.4-4.1 h. Cl and K10 were similar between different doses, respectively. Cmax was 0.93 +/- 0.16 microgram.L-1, Tmax was 2.65 +/- 0.14 h, and elimination T1/2 was 2.5 +/- 0.3 h after s.c. rhGM-CSF. The bioavailability after s.c. rhGM-CSF was 0.61. CONCLUSION: Pharmacokinetics of rhGM-CSF in Macaca mulatta provided a useful index for clinical trial. PMID- 10375760 TI - Effects of 1-(2,6-dimethylphenoxy)-2-(3,4-dimethoxyphenylethylamino) propane hydrochloride on heart function, lactate dehydrogenase and its isoenzymes in rats with cardiac hypertrophy. AB - AIM: To investigate the effects of 1-(2,6-dimethylphenoxy)-2-(3,4 dimethoxyphenylethylamino) propane hydrochloride (DDPH) on cardiac systolic and diastolic function, lactate dehydrogenase (LDH) activity, and LDH isoenzymes in rats with cardiac hypertrophy. METHODS: The cardiac hypertrophy of rats was induced by partly occluding abdominal aorta. The rats were given i.g. DDPH for 8 wk 4 wk after operation, and isolated working heart was made. RESULTS: Eight wk later, in model group, left ventricle systolic pressure (LVSP), LV + dp/dtmax, dp/dtmax and aorta pressure (AP) decreased by 20.2%, 20.0%, 41.4%, and 13.6%, respectively. Left ventricle ending diastolic pressure (LVEDP) increased by 173.9%. The hemodynamic study showed that flowing liquid of aorta (AF) and coronary (CF) and cardiac output (CO) decreased by 49.4%, 41.2%, and 48.9%, respectively. After the rats were given i.g. DDPH, the all above-mentioned parameters recovered to different degrees. Under condition of cardiac hypertrophy, LDH isoenzymes and subunits changed significantly. Isoenzymes LDH3, LDH4, and LDH5, especially LDH5 increased, LDH1 decreased, subunit M in hypertrophied heart increased 1.69 times than that in normal heart. DDPH could decrease subunit M and increase subunit H. CONCLUSION: DDPH can increase cardiac function, coronary flow and reverse changes of LDH isoenzymes in rats with cardiac hypertrophy. PMID- 10375761 TI - Enhancement of ADP-induced aggregation by 5-HT in rabbit platelets. AB - AIM: To study the enhanced effects of 5-hydroxytryptamine (5-HT) on ADP-induced aggregation. METHODS: Platelet aggregation was quantified by the light transmission, the cytosolic-free calcium ([Ca2+]i) was measured by digital fluorescent microscopy, and inositol 1,4,5-triphosphate (IP3) was determined by receptor binding assay. RESULTS: In rabbit platelet-rich plasma (PRP), 5-HT 0.03 3 mumol.L-1 induced a decrease in light transmission (DLT) in a concentration dependent manner with centralization of granules, as revealed by electron microscopy. The DLT was accompanied with neither platelet aggregation nor a release reaction. In single washed platelets loaded with Fura-2, 5-HT caused a concentration-dependent elevation of [Ca2+]i, and IP3 level was also transiently increased in washed platelets at 15 s after stimulation by 5-HT. Adenosine diphosphate (ADP) also caused DLT transiently in PRP before its own aggregation without a release reaction. Pretreatment of PRP or washed platelets with 5-HT, the DLT by ADP was reduced concentration-dependently and ADP-induced aggregation and [Ca2+]i mobilization were enhanced. CONCLUSION: The enhancement of ADP induced aggregation was attributed to the superimposition of the calcium release from the storage sites and calcium influx induced by ADP over the calcium release from the storage sites by 5-HT. PMID- 10375762 TI - Preventive effect of artemether in rabbits infected with Schistosoma japonicum cercariae. AB - AIM: To study the effect of artemether (Art) for prevention of schistosomal infection. METHODS: Rabbits with single infection or reinfection with Schistosoma japonicum cercariae were treated intramuscularly (i.m.) or intragastrically (i.g.) with Art 5 -20 mg.kg-1 on d 7-15 after the first infection, followed by various regimens. RESULTS: When rabbits were injected i.m. Art 7.5 mg.kg-1 (i.e., one half of the effective dose given i.g. on d 7) followed by once every week for twice, the female worm reduction rate was only 42%. In infected rabbits treated i.g. with Art 10-20 mg.kg-1 given in the same administration schedule, the female worm reduction rates were > 91%. When Art 15 mg.kg-1 was given to rabbits on d 7 14 and the following dose of the drug was given at intervals of 7-14 d, the female worm reduction rates were > 94%. In rabbits reinfected with cercariae, the female reduction rate of Art given i.g. once a week for 3 times since d 8 after the first infection was 96% which was similar to that given once a week twice since d 14 after the first infection. CONCLUSION: Art should be given i.g. on d 7 15 after infection, followed by repeated dosing once every 7-15 d for a total of 3 doses. Art given i.g. daily for 2 consecutive days or given at 1-wk intervals since 7-15 d after infection also showed preventive effect. PMID- 10375763 TI - Involvement of interleukin-2 in analgesia produced by Coriolus versicolor polysaccharide peptides. AB - AIM: To study the role of interleukin-2 (IL-2) and mediobasal hypothalamus (MBH) in analgesia produced by Coriolus versicolor polysaccharide peptide (PSP). METHODS: The IL-2 antiserum was injected i.c.v. or i.p. and the MBH was destroyed electrolytically. RESULTS: PSP i.g. 1 g.kg-1.d-1 for 6 d increased the pain threshold in tail stimulation-vocalization test in rats. This PSP-produced analgesia was blocked by i.c.v., but not i.p., IL-2 antiserum and disappeared after electrolytic lesion of MBH. CONCLUSION: The analgesia produced by PSP is mediated by IL-2 which is activated by PSP and interacts with IL-2 receptors in the MBH. PMID- 10375764 TI - Effects of indomethacin on joint damage in rat and rabbit. AB - AIM: To study the effects of indometacin (Ind) on joint damages. METHODS: The volume of noninjected hind paw and interleukin-1 (IL-1) production from peritoneal macrophages and articular synoviocytes induced by lipopolysaccharides were assayed in adjuvant arthritis (AA) rats. Measurements of synovial fibroblast proliferative response and proteoglycan synthesis of cartilage from rabbits were used. RESULTS: The secondary inflammatory reactions in AA rats on d 18, 21, and 24 were suppressed by i.g. Ind 2 mg.kg-1.d-1 for 9 d. Ind promoted IL-1 production from both macrophages and synoviocytes in AA rats. Ind 10 mumol.L-1 enhanced the proliferation of rabbit synovial fibroblasts and suppressed the proteoglycan synthesis of articular cartilage in response to IL-1 in vitro. CONCLUSION: Ind is unfavorable to the repair of joint destruction. PMID- 10375766 TI - Reversal of tumor multidrug resistance by 2-phenyl-3-(3',5'-dimorpholinomethyl-4' hydroxy)-benzoyl-indole (HWL-12). AB - AIM: To explore the reversal of multidrug resistance (MDR) by indole derivative HWL-12. METHODS: Cytotoxicity was determined by tetrazolium (MTT) assay. The function of P-gp was examined by Fura 2-AM assay. Cellular accumulation of doxorubicin (Dox) was measured by fluorescence spectrophotometry. RESULTS: HWL-12 10 mumol.L-1 markedly increased Fura-2 accumulation and was 17.2-fold reversal of MDR in MCF-7/ADR cells. The cellular Dox accumulation in MDR cells was increased in the presence of HWL-12 on the MCF-7/ADR cells. No effect was observed for Dox accumulation in the presence of high Ca2+ (addition of CaCl2) or low Ca2+ (addition of egtazic acid). CONCLUSION: HWL-12 has a potent MDR reversal action which was associated with the increase of cellular Dox accumulation in MDR cells and not related with calcium ion concentration. PMID- 10375765 TI - Fresh vs aged benzylpenicillin on non-IgE responses in mice. AB - AIM: To study whether or not the freshly prepared benzylpenicillin could induce different non-IgE antibody response from aged benzylpenicillin. METHODS: Antibody response was determined by enzyme-linked immunosorbent assay (ELISA). Antigen molecules recognized by antibodies and antigenic cross reactions were tested by hapten inhibition assay. RESULTS: Isotypes of specific non-IgE antibodies induced by freshly prepared benzylpenicillin were mainly IgM, and then IgG and IgA. Some parts of specific antibodies recognized benzylpenicillin molecule and major parts combined with degraded or transforming products. Isotypes of antibodies responsible for cross reaction were mainly IgG between benzylpenicillin and ampicillin and IgM between benzylpenicillin and piperacillin. CONCLUSION: Freshly prepared and aged benzylpenicillin induced different non-IgE antibody response. PMID- 10375767 TI - Effect of catecholamic acid on detoxication and distribution of NiCl2 in mice and rats. AB - AIM: To study the effect of catecholamic acid (CBMIDA) on detoxication of NiCl2. METHODS: Mice and rats were injected s.c. or i.m. CBMIDA immediately after i.p. NiCl2. Each mouse was injected i.p. CBMIDA after i.v. 63NiCl2 185 kBq, and radioactivities of various tissues were measured with liquid scintillation counter at 24 h. The localization of 63Ni was shown by the whole-body autoradiography. RESULTS: CBMIDA s.c. 0.5-1.5 g.kg-1 markedly reduced the mortality from acute poisoning of i.p. NiCl2 500 mg.kg-1. After i.p. NiCl2 in mice, the LD50 was 82.7 mg.kg-1. Mice were injected s.c. CBMIDA 1.5 or 2.5 g.kg-1 after Ni poisoning, the LD50 of NiCl2 were raised to 789 or 820 mg.kg-1, respectively. The LD50 of NiCl2 was 39 mg.kg-1 in rat. If CBMIDA was injected i.m. 0.5 g.kg-1 after i.p. NiCl2, the LD50 was 332 mg.kg-1. CBMIDA 1.5 g.kg-1 i.m. after i.v. 63NiCl2, decreased the contents of 63Ni in blood and lung of mice vs control mice at 24 h. The contents of 63Ni in brain, heart, spleen, and kidney were similar to those of the control mice. The content of 63Ni in bone was more than the control. The excretions of 63Ni through urine and feces were not increased by CBMIDA at 24 h. The whole-body autoradiography showed that the radioactivity was highly localized in the kidney, lung, and Harder's gland. There was a moderate level of 63Ni in the liver, bone, skin, and blood. A pronounced accumulation occurred in the bone. There was a marked reduction of 63Ni in the lung, skin, liver, and blood after i.p. CBMIDA. CONCLUSION: The CBMIDA markedly raised the survival rate of nickel-poisoned mice and rats, and decreased 63Ni levels in lung and blood. PMID- 10375768 TI - Long-term toxicity of modified recombinant human tumor necrosis factor in Macaca mulatta. AB - AIM: To study the long-term toxicity of modified recombinant human tumor necrosis factor (rhTNF-NC) in Macaca mulatta compared with recombinant human tumor necrosis factor (rhTNF). METHODS: rhTNF-NC 93, 9.3 GU/m2, and rhTNF 62 GU/m2 were injected i.v. daily to 16 Macaca mulatta for 1 month and 10 d, respectively. Hematologic, chemical, urinalysis values, ECG, specific antibody, bone marrow, and pathologic profile of organs were measured. RESULTS: No more adverse effects of rhTNF-NC were found in spite of anorexia in 4 monkeys and palpebral edema in 2 monkeys of 93 GU/m2 group. Besides, in rhTNF group, the injury of liver and kidneys, the decrease of erythron, the phlebitis, and thrombosis at injection site occurred. Both drugs caused the production of specific antibody. CONCLUSION: No serious adverse effects of rhTNF-NC were found in Macaca mulatta. The toxicity of rhTNF-NC was much lower than that of rhTNF. PMID- 10375769 TI - [Effect of captopril on platelet cytosolic [Ca2+]i and plasma TXA2/PGI2 in renovascular hypertensive rats]. AB - AIM: To study the effect of captopril (Cap) on platelet cytosolic free calcium concentration ([Ca2+]i), platelet aggregation (PAg), and plasma TXA2/PGI2 ratio in the renovascular hypertensive rats. METHODS: Blood pressure was measured once a week by tail-cuff microphonic manometer. Platelet [Ca2+]i was measured by Fura 2-AM. Plasma angiotensin II (Ang), thromboxane A2 (TXA2), and prostacycline (PGI2) were measured by radioimmunoassay. RESULTS: Platelet [Ca2+]i and PAg increased (P < 0.01), while plasma Ang and TXA2/PGI2 ratio elevated (P < 0.05) in the renovascular hypertensive rats; platelet [Ca2+]i and plasma TXA2/PGI2 ratio reduced markedly after i.g. Cap 100 mg.kg-1.d-1 compared with saline for 2 wk. CONCLUSION: The altered TXA2/PGI2 after Cap treatment contributed to the improvement of the platelet [Ca2+]i and PAg. PMID- 10375770 TI - [Decrease of cAMP and increase of amino acids contents in mouse brain after dihydroetorphine tolerance]. AB - AIM: To study the mechanism of dihydroetorphine (DHE) tolerance. METHODS: DHE tolerance was produced by repeated s.c. injections in progressively increased doses to mice for 8 d. The concentrations of amino acids and cAMP were detected by RP-HPLC/fluorescence assay and radioimmunoassay, respectively. RESULTS: The basal contents of glutamic acid (Glu), aspartic acid (Asp), and GABA in whole brain (cerebellum removed) were increased respectively from 14.1 +/- 2.1, 3.0 +/- 0.4, and 1.8 +/- 0.8 mumol/g tissue in control mice to 17.2 +/- 2.2, 4.1 +/- 0.6, and 3.2 +/- 1.0 mumol/g tissue in tolerant mice, and the rates of increase were 22.0% (P < 0.05), 36.7% (P < 0.01), and 77.8% (P < 0.05 vs control), respectively. There was no significant difference in the basal contents of Gln (5.1 +/- 1.0 vs 4.5 +/- 1.7 mumol/g tissue of control). The basal contents of cAMP in hypothalamus and striatum were decreased respectively from 271 +/- 38 and 189 +/- 31 nmol/g tissue in control mice to 96 +/- 15 and 65 +/- 21 nmol/g tissue in tolerant mice (P < 0.01), and the rates of decrease were 64.6% and 65.6%, respectively. There was no significant difference of cAMP in cerebral cortex (72 +/- 20 vs 55 +/- 15 nmol/g tissue of control). CONCLUSION: The increases of Glu, Asp, and GABA in brain and the decrease of cAMP in hypothalamus and striatum were involved in DHE tolerance. PMID- 10375771 TI - Photodynamic modulation of cellular function. AB - Photodynamic action with a large number of photosensitisers has important practical implications such as photodynamic cancer therapy. But the cellular and molecular mechanisms involved have been rather poorly understood. In this paper, photodynamic modulation of cell signal transduction and the resultant changes in cellular function are reviewed, with a particular emphasis on smooth muscle and the pancreas. PMID- 10375772 TI - Inhibition of fibroblast-like cell proliferation by interleukin-1 blockers, CK 119 and CK-122. AB - AIM: To study potent and nontoxic agents to inhibit fibroblast proliferation. METHODS: Fibroblast-like corneal and conjunctival cells were cultured and inhibited by interleukin-1 (IL-1) blockers, dihydropyridazino-pyridazines CK-119 and CK-122. The cell growth and syntheses of DNA, RNA, and protein after IL-1 blocker incubation were determined. RESULTS: CK-119 and CK-122 inhibited cell growth of corneal fibroblast at 30 mg.L-1. DNA and RNA syntheses in corneal fibroblasts were markedly inhibited by CK-119 and CK-122 whereas protein synthesis was either unaffected or mostly enhanced at 30-100 mg.L-1 and 100-300 mg.L-1, respectively. Similar results were obtained in conjunctival cell cultures by CK-119 and CK-122 at 3-10 mg.L-1 and 30-100 mg.L-1, respectively. CONCLUSION: CK-119 and CK-122 are potent IL-1 blockers to inhibit cell growth of fibroblast like corneal and conjunctival cells mainly through the inhibition of DNA and RNA syntheses but not protein synthesis. PMID- 10375773 TI - Effect of acetylstrophanthidin on action potential duration and relation with extracellular potassium in sheep isolated Purkinje fibers. AB - AIM: To study the relation between the effect of acetylstrophanthidin on action potential duration (APD) and the extracellular potassium concentration. METHODS: Effect of acetylstrophanthidin (AS 0.15 mmol.L-1) on APD at different extracellular potassium concentrations was studied at the stimulation cycle lengths of 990 and 690 ms in sheep isolated cardiac Purkinje fibers using the standard microelectrode technique. RESULTS: At [K+]o 4.0 mmol.L-1, the biphasic effect of AS on APD appeared obviously. Both APD50 and APD90 were lengthened within the first 10 min of drug exposure. After 10 min, they were shortened at all pacing cycle lengths. On the other hand, at [K+]o 5.4 mmol.L-1, AS only shortened APD markedly without lengthening effect on it. The biphasic and monophasic effects of AS on APD were found at [K+]o 4.0 mmol.L-1 and 5.4 mmol.L 1, respectively. CONCLUSION: The effect of AS on APD was related to the concentration of [K+]o. PMID- 10375775 TI - A K+ channel-blocking peptide from venom of Chinese scorpion Buthus martensii Karsch. AB - AIM: To purify and characterize a potassium channel blocker (BmP-3) from the venom of Chinese scorpion Buthus martensii Karsch. METHODS: 1. Purification was carried out by gel-filtration, cation-exchange, and reversed-phase chromatographies. N-terminal was directly sequenced by double-coupling manual method. Molecular weight was determined on an electrospray ionization mass spectrometer. Amino acid composition was analyzed after acidic hydrolysis for 20 h in HCl 6 mol.L-1 at 110 degrees C. 2. Toxicity tests were conducted in mice and cockroaches. 3. The inhibitory effects of BmP-3 on K+ channels were tested in acutely dissociated rat hippocampal pyramidal neurons using whole-cell patch clamp configuration. RESULTS: 1. A pure peptide (BmP-3, 8.1 mg) was obtained, about 0.08% of total proteins of the venom. The N-terminal sequences were VGCEE and the molecular weight was 2938 in ESI-mass spectra. 2. No death occurred at the dosage of 200 micrograms in mice and 8 micrograms in cockroaches. 3. The peptide at 10 mumol.L-1 reduced the peak outward K+ currents by 63% +/- 4% in vitro. CONCLUSION: BmP-3 inhibited K+ channels. PMID- 10375774 TI - Structure-activity relationship of schisandrins in enhancing liver mitochondrial glutathione status in CCl4-poisoned mice. AB - AIM: To explore whether the methylenedioxy group and cyclooctadiene ring of the dibenzocyclooctadiene skeleton of schisandrins (Sch) play a role in the liver mitochondrial glutathione status enhancing activity. METHOD: The effects of three dibenzocyclooctadiene derivatives, Sch A, Sch B, Sch C, and a synthetic intermediate of Sch C, (dimethyl biphenyl dicarboxylate, DBD) on carbon tetrachloride (CCl4)-hepatotoxicity and liver mitochondrial glutathione status were examined in mice. RESULTS: Pretreating mice with intragastric Sch B, Sch C, or DBD 1.mmol.kg-1.d-1 for 3 d protected against CCl4-hepatotoxicity. The hepatoprotection afforded by Sch B or Sch C pretreatment was associated with increases in liver mitochondrial reduced glutathione (mtGSH) level and glutathione reductase (mtGRD) activity, an indication of enhanced mitochondrial glutathione status. In contrast, the hepatoprotective action of DBD was not accompanied by any detectable changes in mtGSH level and mtGRD activity. CONCLUSION: Both the methylenedioxy group and the cyclooctadiene ring of the dibenzocyclooctadiene molecule are important structural determinants in the enhancement of liver mitochondrial glutathione status. PMID- 10375776 TI - Analysis of electronic structures of physostigmine analogs. AB - AIM: To elucidate the action mechanism and structural prerequisites of 21 physostigmine analogs as acetylcholinesterase inhibitors at the molecular level, and help the rational design of these dihydroindoline inhibitors. METHODS: Initial structures of these compounds were built and minimized by SYBYL 6.2 molecular modeling software. Conformations of those molecules with the highest predictive abilities in the Comparative Molecular Field Analysis model were chosen to the semiempirical quantum chemical calculations. RESULTS: (1) The highest occupied molecular orbital (HOMO) consisted mainly of the orbitals in phenyl group and N1 atom; the lowest unoccupied molecular orbital (LUMO) of the molecules was contributed from phenyl group and C11 atom. While the HOMO energies did not show any recognizable relationship with activity, the LUMO energies showed a decreased tendency with increasing activity. The active compounds showed lower LUMO energies. (2) The carbon atom (C11) had the most positive net atom charge. The most active compound had the most positive charge on this carbon, but had the lower charges on the carbonyl oxygen (O12) which was the most negative charge atom. (3) The bond order of carbon-oxygen bond (C11-O10) was invariant across the series of the compounds. (4) Compounds with too high or too low total dipole moment had lower activities, while the most active one had a lower molecular polarizability. CONCLUSION: A molecular model was suggested to explain the possible mode of action by which these compounds inhibit acetylcholinesterase. PMID- 10375777 TI - Dual effects of pentobarbital on rat sacral dorsal commissural neurons in vitro. AB - AIM: To study the effects of pentobarbital (PB) on acutely dissociated rat sacral dorsal commissural neurons (SDCN). METHODS: Nystatin-perforated patch clamp recording was used. RESULTS: (1) At a holding potential of -40 mV, PB induced inward Cl- current (IPB) in a concentration-dependent manner with a EC50 (95% confidence limits) of 416 (385-477) mumol.L-1 and a Hill coefficient of 1.08. (2) Picrotoxin reversibly blocked IPB. (3) The reversal potential of IPB was close to the Cl- equilibrium potential. (4) PB enhanced GABA-induced Cl- influx (IGABA). In the presence of PB 30 mumol.L-1, the EC50 (95% confidence limits) of IGABA decreased from 6.9 (5.4-8.4) mumol.L-1 to 3.5 (2.9-4.1) mumol.L-1. CONCLUSION: PB had dual effects on SDCN, facilitated GABAA receptor-mediated currents and at higher concentrations induced Cl- influx itself. PMID- 10375778 TI - Stimulation of central cholinergic neurons by (-)clausenamide in vitro. AB - AIM: To study the neurotrophic effects of (-) and (+)clausenamide on frontal cortex neurons in culture. METHODS: The activity of the choline acetyltransferase (ChAT) was determined by spectrophotometric method; protein content was assayed by Folin phenol method. RESULTS: (-)Clausenamide increased the activity of ChAT and protein content in cultured neurons, as well as stimulated proliferation of neuronal cells, support survival and neurite outgrowth of neurons. The neurotrophic action of (-)clausenamide (0.001-10 mumol.L-1) was similar to that of nerve growth factor. The (+)clausenamide had no neurotrophic action, even at high concentrations (0.1-10 mumol.L-1), but neurons were damaged. CONCLUSION: ( )Clausenamide stimulated central cholinergic neuron development. PMID- 10375779 TI - Effect of Coriaria lactone on cytosolic free calcium of cultured neurons from rat cerebral cortex. AB - AIM: To study the effect of Coriaria lactone (CL) on cytosolic free calcium ([Ca2+]i) of cultured neurons from cerebral cortex. METHODS: Primary neuron culture (14d) and AR-CM-MIC cation measurement system were used, the [Ca2+]i were measured. CL effect was observed by loading egtazic acid. RESULTS: The [Ca2+]i of cultured neurons (99.4-103.4) nmol.L-1 was elevated concentration-dependently by CL (25-500) mumol.L-1 (P < 0.01). This effect disappeared after loading egtazic acid 5 mmol.L-1, but reappeared after adding CaCl2 to 1 mmol.L-1. CONCLUSION: The [Ca2+]i of cultured neurons was elevated by CL, depending on extracellular Ca2+. PMID- 10375780 TI - Effects of puerarin against glutamate excitotoxicity on cultured mouse cerebral cortical neurons. AB - AIM: To study the effects of puerarin (Pue) against injury of cultured neurons by sodium glutamate (Glu). METHODS: Neuronal damage induced by Glu, N-methyl-D asparate (NMDA), and kainic acid (KA), as well as the actions of Pue and some excitatory amino acid antagonists (EAAA), were measured by determining the leakage of lactate dehydrogenase (LDH) from nerve cells. RESULTS: The 24-h leakage of LDH was increased from cells exposed either to Glu 100 and 500 mumol.L 1 for 15 min (from 20 +/- 4 kU/g protein in control group to 35 +/- 3 kU/g protein in Glu 100 mumol.L-1 group and to 46 +/- 6 kU/g protein in Glu 500 mumol.L-1 group) or to NMDA 500 mumol.L-1 or KA 500 mumol.L-1 for 45 min (from 19 +/- 4 kU/g protein in control group to 27 +/- 3 kU/g protein in NMDA group and to 30 +/- 5 kU/g protein in KA group). Pre and post-treatment with Pue (100 mumol.L 1) decreased the leakage of LDH, which was similar to the effects of EAAA kynurenic acid (from 35 +/- 3 kU/g protein in Glu 100 mumol.L-1 to 20 +/- 5 kU/g protein in kynurenic acid group and to 22 +/- 3 kU/g protein in Pue group), DL-2 amino-5-phosphonovaleric acid (APV) (from 27 +/- 3 kU/g protein in NMDA damaged group to 183 kU/g protein in APV group and to 19 +/- 5 kU/g protein in Pue group) or 6,7-dinitroquinoxaline-2,3(1H,4H)-diane (DNQX) (from 30 +/- 5 kU/g protein in KA damaged control to 22 +/- 5 kU/g protein in DNQX group and to 20 +/- 4 kU/g protein in Pue group). Post-treatment with Pue (100 mumol.L-1) was able to reduce 24-h leakage of LDH from neurons expos ed to Glu 100 mumol.L-1 for 15 min (from 35 +/- 3 kU/g protein to 27 +/- 4 kU/g protein). CONCLUSION: Pue had protective effects on neurons damaged by Glu, NMDA, or KA. PMID- 10375781 TI - Determination of amitriptyline and nortriptyline in human liver microsomes with reversed-phase HPLC in vitro. AB - AIM: To develop a method for simultaneous determinations of amitriptyline (Ami) and its metabolite nortriptyline (Nor) in human liver microsomes. METHODS: An incubation buffer containing microsomes, NADPH-generating system, and Ami, after termination of enzyme reaction and desipramine (Des) as internal standard (IS), was extracted with diethy ether and separated on a reversed-phase ODS column. Detection was achieved at 242 nm by ultraviolet detector. RESULTS: No potential interfering peaks were found. Ami and Nor gave rapid elution and baseline resolution. The linear curves of both analyses ranged 0.02-10 nmol and the limit of detection was 0.01 nmol. The recovery (94%-101%) had good precision with relative s of < 8.3%. CONCLUSION: This method is rapid, sensitive, and simple for studying the metabolism of Ami and Nor. PMID- 10375782 TI - Genetic analysis of N-acetyltransferase polymorphism in a Chinese population. AB - AIM: To study the genetic basis of N-acetylatransferase polymorphism in Chinese. METHODS: Genotypes in 120 healthy Han volunteers from 19 provinces of China were assayed. The 3 common mutant alleles (M1, M2, M3) and one normal wild-type (WT) allele of the N-acetyltransferase (NAT2) gene were detected by allele-specific polymerase chain reaction technique. RESULTS: The NAT2 allele frequencies in 120 Chinese (WT = 0.625, M1 = 0.0458, M2 = 0.188, M3 = 0.142) were different (P < 0.01). The NAT2 genotype distribution for all detected combinations of NAT2 alleles in 120 Chinese subjects was consisitent with Hardy-Weinberg equilibrium (chi 2 = 7.27, nu = 8, 0.7 > P > 0.5). Fifty subjects (41.7%) were homozygous wildtypes, 50 subjects (41.7%) were heterozygous mutants, and 20 subjects (16.7%) were homozygous mutants. CONCLUSION: The lower frequency of mutant M1 allele compared with that of Caucasians explains the low frequency of slow acylators in Chinese. PMID- 10375783 TI - Effects of vitamin C on myocardial mitochondrial function and ATP content in hypoxic rats. AB - AIM: To observe the effects of large dose of vitamin C (Vc) on myocardial mitochondrial function, ATP content, and myocardial structure in acute and chronic hypoxic rats. METHODS: Rats were exposed to a simulated altitude 4000 m (barometric pressure = 43 kPa) for 3 and 30 d. Vc (0.75 g.kg-1.d-1) was injected i.p. The heart mitochondrial respiratory function were determined by Clark-type O2 electrode; mitochondrial membrane fluidity (MMF) were assayed through fluorescence polarizative method; the contents of ATP, ADP, and AMP in myocardial tissue were measured with HPLC. RESULTS: After administration of Vc, the ATP content was increased from 35 +/- 3 mg.g-1 to 53 +/- 3 mg.g-1 in acute hypoxic rats (P < 0.01), from 42 +/- 4 mg.g-1 to 48 +/- 3 mg.g-1 in chronic hypoxic rats (P < 0.01); Pa, O2 was increased from 7.2 +/- 1.4 kPa to 9.5 +/- 1.2 kPa in acute hypoxic rats (P < 0.01); mitochondrial respiratory control rate (RCR) was increased from 2.1 +/- 0.6 to 4.7 +/- 0.5 in acute hypoxic rats (P < 0.01), and from 3.3 +/- 0.7 to 4.5 +/- 0.6 in chronic hypoxic rats (P < 0.01); MMF was increased in acute and chronic hypoxic rats (P < 0.05); the degree of myocardial necrosis in vitamin C preventive rats was attenuated as compared with those of acute hypoxic rats. CONCLUSION: Vc is effective on improving myocardial energy metabolism and protecting against myocardial structural injury in hypoxic rats. PMID- 10375784 TI - Effect of mitoxantrone on DNA polymerase of Ehrlich ascites carcinoma cells. AB - AIM: To study the effect of mitoxantrone (Mit) on DNA polymerases of tumor cells. METHODS: DNA polymerases of Ehrlich ascites carcinoma cells were isolated by phosphocellulose column chromatography. The effects of Mit on DNA polymerase alpha, beta, and gamma were detected by method of K Ono. RESULTS: Mit inhibited DNA polymerase alpha, beta, and gamma, IC50 values were 11.9, 6.5, and 11.9 mumol.L-1, and Ki 1.86, 2.22, and 2.05 mumol.L-1, respectively. The inhibitory mode of Mit on DNA polymerase alpha, beta, and gamma was competitive. CONCLUSION: Mit is a strong inhibitor on DNA polymerase alpha, beta, and gamma. The inhibitory mode was competition with respect to template DNA. PMID- 10375785 TI - Protective effects of Ginkgo biloba extract against lysophosphatidylcholine induced vascular endothelial cell damage. AB - AIM: To study the protective effects of Ginkgo biloba extract (GbE) against endothelial cell damage induced by lysophosphatidylcholine (LPC). METHODS: The vasorelaxation response to acetylcholine (ACh) were investigated in the isolated rabbit thoracic aorta. Lipid peroxidation products were determined by measuring thiobarbituric acid reactive substance. RESULTS: GbE attenuated the inhibition of vasorelaxation response to ACh and prevented the LPC-induced increase of malondialdehyde (MDA) content both in thoracic aortae. GbE prevented the leakage of LDH and the increase of MDA content in cultured endothelial cells in a concentration-dependent manner. GbE also markedly increased epoprostenol level in cultured endothelial cells treated with LPC. CONCLUSION: GbE protected endothelial cells against LPC-induced damage due to reduction in lipid peroxidation and facilitation of synthesis and/or release of epoprostenol. PMID- 10375786 TI - Effects of amiodarone on cardiac electrophysiology in right ventricular rapid pacing-induced heart failure dogs. AB - AIM: To study the effects of amiodarone (Ami) on cardiac electrophysiologic properties and ventricular fibrillation threshold (VFT) in right ventricular rapid pacing-induced congestive heart failure (CHF) dogs. METHODS: Dogs (n = 25) were randomly allocated into 3 groups: A) control group; B) CHF group induced by right ventricular rapid pacing (4 pulses.s-1) for 4-5 wk; C) CHF models p Ami 300 mg.d-1 for 4-5 wk. The electrophysiologic parameters and VFT were evaluated by electric stimulation and monophasic action potential (MAP) recording. RESULTS: In CHF models, ventricular MAP duration (MAPD90), ventricular late repolarization duration (VLRD), and intra-ventricular conduction time (IVCT) were prolonged by 43%, 318%, and 19%, respectively; the ratio of ventricular effective refractory period (VERP) to MAPD90 (VERP/ MAPD90) and VFT were decreased by 13% and 48% respectively; the dispersion of ventricular recovery time (RT-D) was increased by 185%. In CHF models, Ami had no effects on ventricular MAPD90, but increased VERP/ MAPD90, IVCT, and VFT by 15%, 10%, and 67%, respectively, shortened VLRD by 87%; and decreased RT-D by 87%. Ami had no significant influences on the hemodynamic parameters of the CHF dogs. CONCLUSION: Ami normalizes the cardiac electrophysiologic properties in CHF dogs. PMID- 10375787 TI - Modulating effect of mitomycin or cisplatin on lymphokine-activated killer cell proliferation and antitumor activity to bladder cancer cell lines in vitro. AB - AIM: To study the effect of mitomycin (Mit) or cisplatin (Cis) on the proliferation of lymphokine-activated killer (LAK) cells in patients with transitional cell cancer of bladder and their cytolysis to bladder tumor cells. METHODS: LAK cell proliferation was assayed in the presence of Mit or Cis by cell counting. Bladder cancer cell lines BIU-87 and EJ were cultured as target cells and cytotoxicity of LAK cells was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay. RESULTS: The proliferation of LAK cells induced by recombinant interleukin-2 (IL-2) was inhibited by Cis in a concentration-dependent manner and was decreased to 55.3% at 100 mg.L-1 compared with control at 96 h. The enhanced growth of the LAK cells was observed with Mit 5-10 mg.L-1 from 48 to 96 h. Cis 10 mg.L-1 increased the cytotoxicity against BIU 87 and EJ cells. CONCLUSION: Immunomodulatory effect of chemotherapeutic agents on LAK cell proliferation induced by IL-2 in patients with bladder cancer mainly depends on the drug itself. PMID- 10375788 TI - Effects of ciclosporin on whole blood chemiluminescence of renal transplant patients. AB - AIM: To examine the possible inhibitory role of ciclosporin (Cic) on luminol dependent chemiluminescence (CL) of whole blood in renal transplant patients. METHODS: Luminol-dependent CL was used to measure active oxygen species generation in respiratory burst of whole blood stimulated by zymosan A. Fluorescence polarization immunoassay was used to monitor the blood concentration of Cic. RESULTS: CL values of Cic group (n = 50) decreased in comparison with those of normal group (n = 10) (P < 0.01). The blood concentration of Cic was negatively related to CL value (P < 0.01). The serum of renal transplant patients directly inhibited respiratory burst of peritoneal macrophages of rats in a concentration-dependent manner. CONCLUSION: Cic inhibits the phagocytic activity of neutrophils in renal transplant patients. PMID- 10375789 TI - Effect of procainamide on ultrastructure of blood platelet in rabbits. AB - AIM: To study the effect of procainamide (PA) on the ultrastructure of blood platelets. METHODS: Arachidonic acid was added to PA-treated platelet-rich plasma to induce platelet aggregation. The 50-nm sections were examined with a transmission electron microscope. RESULTS: PA 8.5-136 mumol.L-1 markedly inhibited changes of pseudopods, alpha-granules, dense granules, glycogens, open canalicular system, and dense tubular system. CONCLUSION: PA markedly inhibited the changes of ultrastructure of blood platelet and releasing response. PMID- 10375790 TI - Platelet-released ADP stabilizes PAF-induced rabbit platelet aggregation by stabilizing intracellular calcium. AB - AIM: To examine whether platelet-released adenosine diphosphate (ADP) would contribute to the stabilization of rabbit platelet aggregation induced by platelet activating factor (PAF). METHODS: Rabbit platelet aggregation induced by PAF was measured turbimetrically. ADP release from rabbit platelets stimulated by PAF was determined by HPLC. Intracellular Ca2+ was measured using Ca(2+) sensitive fluorescent indicator Fura 2-AM. RESULTS: PAF > or = 1 nmol.L-1 induced full platelet aggregation, which did not deaggregate over 5 min after aggregation reached peak. Platelet aggregation was deaggregated in a concentration-dependent manner by subsequent addition of ADP scavenger ATP-diphosphohydrolase (apyrase) at 5-100 mg.L-1. PAF 3 nmol.L-1 stimulated release of ADP (29% vs 6% of control), and elicited a rapid rise in intracellular calcium ([Ca2+]i) which peaked at approximately 15 s. Then the [Ca2+]i gradually decayed from 585 +/- 80 nmol.L-1 within 100 s to a low level (364 +/- 82 nmol.L-1). Apyrase 100 mg.L-1, added 2 min after PAF, reduced [Ca2+]i to a lower level (171 +/- 29 nmol.L-1). CONCLUSION: Platelet-released ADP stabilizes PAF-induced rabbit platelet aggregation by stabilizing [Ca2+]i at elevated level. PMID- 10375791 TI - Effects of 5-HT released from platelets on thrombin-induced aggregation and ATP release in rabbit platelets in vitro. AB - AIM: To study the effects of arachidonic acid (AA)-induced endogenous serotonin (5-HT) release on platelet aggregation and ATP release by thrombin (Thr). METHODS: Platelet aggregation and release reaction were quantified by light transmission in platelet-rich-plasma (PRP) and the amount of ATP in medium. The effects of endogenous 5-HT were evaluated by the filtration of content in cuvette A (content A) containing endogenous 5-HT into cuvette B in which Thr-induced aggregation was observed in the absence/presence of ?(+/-)-5 (Z)-7-[3 endophenylsulfonylamino [2.2.1] bicyclohept-2-exo-yl]heptanoic acid, sodium salt? (S-145) or/and methysergide (Met). RESULTS: (1) AA 100 and 200 mumol.L-1 induced aggregation and ATP release in cuvette A. When the aggregation reached a peak, the content A directly caused platelet aggregation in cuvette B, and it was inhibited by S-145 100 nmol.L-1, Met 30 mumol.L-1, and inhibited more potently by S-145 + Met. (2) In the presence of S-145 100 nmol.L-1 in cuvette B, aggregations by Thr 0.1 and 0.3 IU.L-1 were enhanced (P < 0.01) by the filtrate, while Thr 0.5 IU.L-1-caused ATP release was suppressed (P < 0.01) without the effect on aggregation. Preincubation with S-145 and Met, the effects of the filtrate on aggregation and ATP release were abolished. (3) By prolongation of the time intervals between filtration and addition of Thr, the aggregation was enhanced and ATP release was reduced. CONCLUSION: Endogenous 5-HT was released from activated platelet and plays, in turn, a role in the regulation of platelet aggregation by the superimposition of cytosolic-free calcium ([Ca2+]i) and the feedback loop to regulate release reaction and calcium. PMID- 10375792 TI - Immunosuppressive effects of intravenous self administration of dihydroetorphine on lymphocyte functions in rats. AB - AIM: To study the effects of dihydroetorphine (DHE) on lymphocyte functions in rats and to further assess the abuse potential of DHE. METHODS: An intravenous self administration (SA) procedure in rats was used to determine the SA liability of DHE. Concanavalin A (Con A)-stimulated lymphocyte proliferation and lymphokine production of rat splenocytes were measured. RESULTS: DHE 178 +/- 13 micrograms established a stable and typical rat model of psychological dependence, suppressed lymphocyte proliferation (129 +/- 11 Bq) compared with control group (620 +/- 36 Bq), and inhibited the activity of interleukin-2 (IL-2) (A570 = 0.28 +/- 0.06) compared with control group (A570 = 0.51 +/- 0.03). CONCLUSION: DHE had a high abuse potential and inhibited the Con A-induced lymphocyte proliferation and interleukin-2 production in rats. PMID- 10375793 TI - Effects of nitroquine on ultrastructures and cytochrome oxidase of exoerythrocytic Plasmodium yoelii in rat liver. AB - AIM: To study the effects of nitroquine acetate (NA) on the ultrastructures and cytochrome-c oxidase (CCO) of exoerythrocytic forms (EEF) of Plasmodium yoelii. METHODS: Rats were inoculated with sporozoites directly into the liver. After 48 h rats were killed. Rat liver thin sections were incubated in histochemical reaction medium, then examined by transmission electron microscopy. NA (2 mg.kg 1) was fed to rats 3.5 h and 14 h before killing the rats. RESULTS: At 3.5 h, in the parasites there appeared swelling and proliferation of mitochondria, dilation of endoplasmic reticulum, and reduction of the electron density of parasites' nuclei. The structures of the parasites disintegrated to form many autophagocytes 14 h after exposure to NA. The reaction products of CCO still existed until 14 h after using NA. CONCLUSION: CCO was not the starting point of NA action. NA interferes with the structure and function of the cytoplasm and nucleus of malaria parasites and exerts its antimalarial effects in many aspects. PMID- 10375794 TI - [Effect of corticotropin on formalin-evoked c-fos expression of spinal cord and receptors analysis in rat]. AB - AIM: To study the effect of corticotropin (Cor) on c-fos expression induced by formalin in spinal cord of rat and the role of receptors. METHODS: Using immunohistochemistry and adrenalectomy. RESULTS: Cor inhibited the formalin evoked c-fos expression in rat spinal cord in a dose-dependent manner. No obvious effect was seen by i.p. Cor 10 U.kg-1, but 25 U.kg-1 reduced the evoked c-fos expression, that was blocked by phentolamine, naloxone, or verapamil, but not much changed by adrenalectomy. CONCLUSION: The formalin-evoked c-fos expression of rat spinal cord was suppressed by Cor through the alpha-adrenergic receptors, opiate receptors and calcium, but no relation to adrenal glands. PMID- 10375795 TI - Cardiovascular ATP-sensitive K+ channel as a new molecular target for development of antihypertensive drugs. PMID- 10375796 TI - Endomorphin-1 and -2 inhibit human vascular sympathetic norepinephrine release: lack of interaction with mu 3 opiate receptor subtype. AB - AIM: To determine if endomorphin-1 (End-1) and -2 (End-2) interact with mu 3 opiate receptor subtype and in this way cause vascular hypotension. METHODS: Amperometric nitric oxide (NO) determinations associated with opiate binding displacement analysis and preloaded [3H]norepinephrine KCl stimulated release in human vascular tissues from sympathetic nerve fibers in vitro. RESULTS: The endomorphins did not release NO from human monocytes, granulocytes, saphenous vein, and internal thoracic artery endothelium and did not displace opiate alkaloid binding to mu 3 receptor. However, they did inhibit KCl-stimulated [3H]norpinephrine release from vascular nerves. CONCLUSION: The data strongly suggested that End-1 and -2 caused hypotension by blocking sympathetic vascular sympathetic activity. PMID- 10375797 TI - Wiedendiol-A inhibits cholesteryl ester binding to its transfer protein. AB - AIM: To study the wiedendiol-A (W-A) inhibition mechanism of plasma cholesteryl ester (CE) transfer protein (CETP) on the transfer of CE. METHODS: Using gel filtration method. RESULTS: W-A at 30 mumol.L-1 inhibited association of CE with CETP by 76% and CETP transfer activity by 81%. In addition, W-A enhanced binding of TP2, a monoclonal antibody with a CETP C-terminal epitope which is involved in CE binding, to CETP, suggesting a W-A-induced conformational change at the epitope for increased TP2 binding. When CETP activity was measured by varying high-density lipoproteins (HDL) concentration, the apparent Vmax of CE transfer was inhibited by 74% and 83% in the presence of W-A at 14 and 25 mumol.L-1, respectively, while the apparent K(m) of HDL for CETP did not change. CONCLUSION: W-A action is mediated through interaction between W-A and CETP, but not through those between W-A and lipoproteins. PMID- 10375798 TI - Effects of synthetic (-)-huperzine A on cholinesterase activities and mouse water maze performance. AB - AIM: To compare the effects of synthetic and natural (-)-huperzine A (Hup A) on cholinesterase and mouse water maze performance. METHODS: Spectrophotometry was used to determine cholinesterase activity. Mouse water maze was used to evaluate nootropic effect. RESULTS: The IC50 of synthetic Hup A for acetylcholinesterase (AChE) of rat cortex and rat erythrocyte membrane determined in vitro were 64.7 (52.6-79.5) and 53.9 (43.6-66.6) nmol.L-1, respectively, and for butyrylcholinesterase of rat serum was 53.6 (44.9-63.8) mumol.L-1. Synthetic Hup A 0.12-0.48 mg.kg-1 ig produced a dose-dependent inhibition of brain AChE in mice. Synthetic Hup A 0.05 mg.kg-1 ig attenuated scopolamine-induced impairment of spatial memory. The efficacy of synthetic Hup A was the same as natural Hup A. CONCLUSION: Synthetic Hup A yielded an in vitro and in vivo pharmacological profile of activities similar to that of natural Hup A. PMID- 10375799 TI - Hemorheology and walking of peripheral arterial occlusive diseases patients during treatment with Ginkgo biloba extract. AB - AIM: To study the effects of Ginkgo biloba extract 761 (GbE) from the points of view of hemorheology for patients of peripheral arterial occlusive diseases (PAOD). METHODS: The treatment with GbE (240 mg.d-1, po) and the pain-free walking distance (PFWD) were carried out for 24 PAOD patients (12 nondiabetic, ND and 12 diabetic, D) over 48 wk. The parameters erythrocyte stiffness (ES) and relaxation time (RT), the blood plasma viscosity (eta), the plasma fibrinogen concentration (Cf) and the blood sedimentation rate (BSR), the PFWD, and maximal walking distance (MWD) were determined at 6 wk before treatment (-6), at the beginning of the treatment (0), and after 6, 11, 16, and 48 wk of treatment. RESULTS: At wk -6, ES and RT of both the ND- and D-group were not significantly different from a healthy control group. At wk 0, stiffness and RT were significantly higher than healthy control, and the mean PFWD was only 111 m. The eta value was significantly elevated and Cf and BSR were enhanced. Throughout 11 wk of treatment ES, RT, eta, and Cf decreased gradually and PFWD improved. Between 16 and 48 wk, ES, and RT were no longer significantly different from the controls, whereas eta and Cf decreased gradually but remained higher than normal, BSR decreased, and the PFWD improved by a factor of 3.8 times (D) and 3.3 times (ND). CONCLUSION: GbE gives therapeutic effects in PAOD patients. PMID- 10375800 TI - Intradermal irritation of dental bleaching agents in rats. AB - AIM: To evaluate the irritative potential of three dental bleaching agents (hydrogen oxide, carbopol, and carbamide peroxide). METHODS: In rats, Evans blue (2.5%, 1 mL.L-1) was injected i.v. and later each test solution was injected intradermally on the back. After the concentration of the dye in the stained skin area was determined by spectrophotometric analysis. RESULTS: All the dental bleaching agents caused increase of vascular permeability and the intensity varied with the time. CONCLUSION: Dental bleach agents had a great potential for irritating soft tissues. PMID- 10375801 TI - Cytotoxicities of alkaloids from processed and unprocessed seeds of Strychnos nux vomica. AB - AIM: To examine the cytotoxicities of 6 crude Strychnos alkaloid fractions from the seeds of Strychnos nux-vomica unprocessed or processed with various traditional processing methods and 13 pure Strychnos alkaloids from the fractions. METHODS: Using cell culture, their inhibitory effects on Vero cell growth-inhibition assay, and host cell DNA synthesis by [3H]thymidine ([3H]TdR) uptake assay. RESULTS: The IC50 of processed seeds were 155% and 212% of unprocessed ones in cell growth-inhibition assay and in [3H]TdR uptake assay, respectively. The IC50 of 13 compounds were 0.45-0.80 mmol.L-1 and 0.50-12 mmol.L 1, respectively. The processing method with sand bath exhibited a wide safety margin compared with other traditional processing methods or no processing. The isomers of Strychnos alkaloids and their N-oxides showed much lower cytotoxicities among these alkaloids. Isobrucine N-oxide showed the lowest cytotoxicity. The contents of isomers and N-oxides of Strychnos alkaloids were the highest in the sand processing. CONCLUSION: Processing of nux vomica plays a critical role in its toxicity. PMID- 10375802 TI - An expressional system of human cytochrome P-450 CYP1A1 gene transcription. AB - AIM: To explore an expressional system of human cytochrome P-450 CYP1A1 (CYP1A1) gene transcription. METHODS: The plasmid pMC 6.3 K containing human CYP1A1 promoter was transiently transfected into Hep G2 cells. The expression of chloramphenical acetyltransferase (CAT) reporter gene was detected by ELISA. RESULTS: Both the CAT expression and CYP1A1 activity increased with the concentrations of beta-naphthoflavone from 2.5 to 10 mumol.L-1. At 10 mumol.L-1 of beta-naphthoflavone, the levels of CAT and CYP1A1 were 94-fold and 2.8-fold those of the corresponding control, respectively. Using this method, the study of 8 glucosinolates with various side chains on the induction of CYP1A1 gene transcription showed that none of the parent glucosinolates increased CAT expression, whereas the breakdown products of indol-3-yl-methyl glucosinolate (glucobrassicin), rather than indole-3-carbinol, increased the CAT expression. CONCLUSION: The CYP1A1 gene transcriptional system was more reliable and sensitive. PMID- 10375803 TI - Clomipramine N-demethylation metabolism in human liver microsomes. AB - AIM: To study the effect of cytochrome P-450 (CYP450) inhibitors on clomipramine (Clo) N-demethylation in vitro. METHODS: The kinetic parameters of Clo N demethylation in human liver microsomes were obtained by the Michaelis-Menten equation. The parameters after pretreatment with putative inhibitors of various CYP450 isoforms were compared with controls. RESULTS: K(m1), K(m2), Vmax1, Vmax2, Vmax1/K(m1), and Vmax2/K(m2) were (0.11 +/- 0.06), (24 +/- 14) mumol.L-1, (114 +/ 47), (428 +/- 188) nmol.g-1.min-1, (1.8 +/- 1.6), and (0.019 +/- 0.005) L.g 1.min-1, respectively. The interindividual variations for the last 4 parameters reached up to 2.5-, 7.3-, 3.4-, and 1.8-fold. At 5 mumol.L-1 of Clo, troleandomycin (Tro), furafylline (Fur), ditiocarb sodium (Dit), and S mephenytoin (Mep) produced a marked inhibition on Clo N-demethylation while sulfaphenazole (Sul) and quinidine (Qui) had only slight effects. The inhibitory rates by Dit 30, Mep 500, Fur 10, Tro 10, Fur 80, Tro 200 and Fur 80 + Tro 200 mumol.L-1 were 27.0%, 32.9%, 42.8%, 40.5%, 63.9%, 66.4%, and 78.3%, respectively. The IC50 (95% confidence limits) for Fur and Tro were 27.7 (19.1-36.3) and 42.1 (20.9-63.3) mumol.L-1, respectively. CONCLUSIONS: The N-demethylation of Clo exhibited a biphasic behavior. This reaction was mediated mainly by both CYP1A2 and CYP3A4, to a minor extent by CYP2C19 at the low concentration of Clo in vitro. PMID- 10375804 TI - Pharmacokinetics of theophylline metabolites in 8 Chinese patients. AB - AIM: To study theophylline metabolites pharmacokinetics in patients after a therapeutic dose. METHODS: Eight adult patients with mild bronchial asthma and normal liver function were infused aminophylline intravenously (6.6 mumol.kg-1). The plasma concentrations of theophylline and its 4 metabolites: 1,3-dimethyluric acid (DMUA), 3-methylxanthine (3-MX), 1-methyluric acid (MUA), and the intermediate 1-methylxanthine (1-MX) were monitored by HPLC throughout 24 h. RESULTS: The plasma concentration of DMUA was the highest one among the 4 metabolites. 3-MX showed the slowest elimination rate. The plasma concentration of 1-MX throughout a 24-h period showed that there was a picking up of 1-MX (from 0.04 mumol.L-1 to 1.05 mumol.L-1) in the next morning. CONCLUSION: The formation of DMUA was the main metabolites. During night there was an accumulation of 1-MX. PMID- 10375805 TI - Stereoselective metabolism of metoprolol in isolated rat liver. AB - AIM: To study the stereoselective pharmacokinetics of the enantiomers of metoprolol (Met). METHODS: The enantiomers of Met were metabolized in isolated rat liver with hemoglubin-free medium. The enantiomers of Met were analyzed with HPLC. RESULTS: The linear kinetics were found in 3 doses (0.16, 0.32, and 0.64 mg) of R-(+)- and S-(-)-Met. The time-concentration curve of Met was fitted in first order-kinetics compartment model. There were differences in the pharmacokinetic parameters (K, T1/2, and Cl) between the enantiomers at the same dose (P < 0.01). The ratio of liver clearence of S-(-)-Met/R-(+)-Met was 0.14 0.17. CONCLUSION: Pharmacokinetics of enantiomers of Met in the isolated rat liver were stereoselective, with a preferential clearance of R-(+)-Met, which elimination was not a saturable process. PMID- 10375806 TI - Delay of metabolism rate of ciclosporin by simvastatin in 7 Chinese healthy men. AB - AIM: To study the effects of simvastatin (Sim) on pharmacokinetics of ciclosporin (Cic). METHODS: Seven healthy young volunteers took Cic 100 mg alone or in combination with Sim 10 mg in a randomized crossover study. The Cic concentrations in blood were determined by specific fluorescence polarization immunoassay. Data were analyzed with 3P87 program. RESULTS: The blood concentration-time curve was fitted to open 2-compartment model, and the pharmacokinetic parameters of Cic alone and Cic + Sim were: Cmax (646 +/- 94) and (698 +/- 340) micrograms.L-1; Tmax (1.12 +/- 0.13) and (1.13 +/- 0.21) h; AUC (2.3 +/- 0.4) and (2.6 +/- 1.2) mg.h.L-1; T1/2 beta (12 +/- 6) and (23 +/- 8) h (P < 0.05). CONCLUSION: Sim delays the metabolism rate of Cic when they are given simutaneously. PMID- 10375807 TI - Establishment of kappa opioid receptor agonists pharmacophore with molecular modeling method. AB - AIM: To build up nonpeptide kappa-opioid receptor agonists pharmacophore. METHODS: Five structurally diverse, highly active nonpeptide kappa-opioid agonists were retrieved from MDL MDDR database. Molecular mechanics method were used to seek out 50 lower energy conformations for each compound. Taking nitrogen atom of pyrrolidine and acyl acetamide as overlay points, 5 agonists were superimposed to each other with their most favorite conformation. From the overlay map, the structure specificity of nonpeptide kappa-opioid receptor agonists were elucidated. RESULTS: According to this pharmacophore, the pyrrolidine ring, the carbonyl group of acyl acetamide, and the hydrophobic group attached to acyl acetamide were suggested to be the structure-specific moieties of kappa-opioid agonists. Moreover, by comparing kappa 1-opioid receptor sequence of mouse with other G-protein-coupled receptors, we determined those conserve residues existing on transmembrane regions which might interact with the suggested groups. The carboxyl of Asp138 might interact with N atom of pyrrolidine by forming a hydrogen bond. The hydroxyl of Ser187 and the carbonyl group of kappa-opioid agonists might form another hydrogen bond, which was critical for its kappa selective affinity. The hydrophobic group attached to acyl acetamide might have hydrophobic interaction with aromatic residues of kappa opioid receptors. CONCLUSION: These kappa agonists pharmacophore were helpful to select specific positions in the lead compounds to be occupied by hydrophobic moieties to limit their ability to across the blood-brain barrier. PMID- 10375808 TI - Tetraethylammonium and 4-aminopyridine enhancement of 5-HT3 receptor-mediated contraction of guinea pig ileum in vitro. AB - AIM: To study the effects of tetraethylammonium (TEA) and 4-aminopyridine (4-AP) on 5-HT3 receptor-mediated contractions of the isolated guinea pig ileum longitudinal muscle-myenteric plexus strip preparations (GPI). METHODS: GPI contractions were recorded with a chart recorder through isometric transducers. The effect of TEA and 4-AP on binding properties of 5-HT3 receptors was assessed using [3H]GR65630 binding assay in membrane preparation of rat entorhinal cortex. RESULTS: (1) Both TEA 0.5 mmol.L-1 and 4-AP 5 mumol.L-1 increased the spontaneous activity, and elicited contractions of GPI; atropine 10 mumol.L-1 or the selective 5-HT3 receptor antagonist MDL72222 100 mumol.L-1 prevented these effects. (2) Both TEA 0.05-0.5 mmol.L-1 and 4-AP 1-10 mumol.L-1 enhanced GPI contractions induced by the selective 5-HT3 receptor agonists 2-methyl-5-HT in concentration-dependent manners. (3) Both TEA 0.5 mmol.L-1 and 4-AP 5 mumol.L-1 attenuated the inhibitory effects of the selective 5-HT3 receptor antagonists tropisetron 0.1 mumol.L-1 and benesetron 1 mumol.L-1 on 5-HT3 receptor-mediated GPI contractions. (4) Neither TEA 0.1-0.5 mmol.L-1 nor 4-AP 5-10 mumol.L-1 affected GPI contractions evoked by the selective M-ACh receptor agonist carbachol 1 mumol.L-1. (5) TEA 0.5 mmol.L-1 and 4-AP 10 mumol.L-1 had no effect on the properties of binding of the selective 5-HT3 receptor radioligand [3H]GR65630 to 5-HT3 receptors. CONCLUSION: The enhancement by TEA and 4-AP of 5 HT3 receptor-mediated GPI contractile responses was due to blocking K+ channels in prejunctional myenteric neurons. PMID- 10375809 TI - L-type calcium channel blockade mechanisms of panaxadiol saponins against anoxic damage of cerebral cortical neurons isolated from rats. AB - AIM: To identify the changes of L-type Ca2+ channel on cerebral cortical neurons of rats during anoxia and the protective mechanisms of panaxadiol saponins (PDS) against anoxic injury. METHODS: Patch-clamp technique of cell-attached configuration and in vitro cerebral anoxic modle built with actuely isolated cortical cells of Wistar rats. RESULTS: The open time of L-type Ca2+ channel of cortical neurons increased significantly from (2.85 +/- 0.21) ms to (9.1 +/- 1.0) ms (P < 0.01) under anoxia. The particular change was a long-lasting open, which was more than 20 ms in some cases. At the same time, the close time decreased from (38 +/- 8) ms to (10 +/- 3) ms (P < 0.01) and the open-state probability raised from (0.047 +/- 0.008) to (0.165 +/- 0.025) (P < 0.01). PDS (1.5 g.L-1) inhibited the activity of L-type Ca2+ channel both in normal and anoxic condition [open time from (2.23 +/- 0.47) ms and (9.1 +/- 1.0) ms to (1.03 +/- 0.25) ms and (2.1 +/- 0.4) ms; close time from (38 +/- 10) ms and (10 +/- 3) ms to (74 +/- 16) ms and (46 +/- 10 ms); open-state probability from (0.043 +/- 0.006) and (0.165 +/- 0.025) to (0.012 +/- 0.004) and (0.021 +/- 0.009), respectively, P all < 0.01]. The results of PDS were similar to those of verapamil, but were weaker compared with verapamil. CONCLUSION: The L-type Ca2+ channels of rat cerebral cortical neurons were obviously opened during anoxia. The channels in normal and anoxic condition were effectively blocked by PDS. It was one of the important mechanisms by which PDS protected brain from the anoxic injury. PMID- 10375810 TI - Intracerebroventricularly injected L-arginine-induced vasopressin release is mediated by cGMP in rats. AB - AIM: To study the possible role of cGMP in the effect of L-arginine-induced arginine-vasopressin (AVP) release. METHODS: In anesthetized rats, saline, L arginine, 8-Br-cGMP, L-arginine + methylene blue (an inhibitor of guanylyl cyclase) were injected intracerebroventricularly (icv) and the plasma vasopressin level was detected with radioimmunoassay. RESULTS: Both 8-Br-cGMP 2.53 g.L-1 and L-arginine 100 g.L-1 increased plasma AVP level [from (2.6 +/- 0.3) to (6.6 +/- 0.4) ng.L-1, and from (3.2 +/- 0.5) to (5.8 +/- 1.4) ng.L-1, respectively; P < 0.01] 5 min after the icv injection; methylene blue (3.74 g.L-1) + L-arginine (100 g.L-1) did not change plasma AVP level. CONCLUSION: cGMP was a mediator of the effect of L-arginine-induced AVP release. PMID- 10375811 TI - Up-regulation of LPS-induced iNOS activity in dibutyryl cyclic AMP-differentiated rat astrocytes. AB - AIM: To study the effect of dBcAMP on bacterial endotoxin LPS-induced NOS activity. METHODS: Microscopic changes were observed. Nitrite levels were measured by fluorometric assay. NOS activity was measured by citrulline assay. RESULTS: Within 3-4 h after the addition of dBcAMP 1 mmol.L-1 to culture medium, a morphological transformation reminiscent of in vivo differentiation occurred. Coincubation with LPS and dBcAMP 1 mmol.L-1 resulted in a marked increase in the nitrite production as compared with LPS alone. This increase was concentration- and time-dependent with a maximal effect after 24 h treatment. Nitrite production stimulated by LPS is parallel to the degree of cell differentiation. After a 24-h costimulation with LPS and dBcAMP, L-citrulline formation assay revealed a 3-fold increase in NOS activity over LPS treatment alone. Simultaneous incubation with L NAME, completely inhibited the stimulation effect of LPS/dBcAMP on nitrite production. Cycloheximide and dactinomycin also suppressed enhancement of NOS activity stimulated by LPS/dBcAMP, both in nitrite production and citrulline assay, indicating that the enhancement of NOS activity was due to the expression of inducible NOS (iNOS) gene and protein. CONCLUSION: Inflammatory signals can trigger astrocytes to express substantially different levels of iNOS depending on their degree of differentiation. PMID- 10375812 TI - Inhibitory effects of ginkgolides on nitric oxide production in neonatal rat microglia in vitro. AB - AIM: To study the effects of ginkgolide A (GA) and B (GB), apafant (Apa), and N omega-nitro-L-arginine (L-NA) on nitric oxide (NO) production in cultured neonatal rat microglia. METHODS: NO concentration was represented by nitrite which was determined by Griess reaction. RESULTS: In resting microglia, no inhibitory effects of GA, GB, and Apa were observed. L-NA inhibited NO production, its IC50 value (95% confidence limits) being 3.4 (0.8-14.9) mumol.L 1. GA, GB, and L-NA inhibited NO production in LPS-stimulated microglia, and their IC50 values (95% confidence limits) were 5.7 (1.8-18.1), 1.1 (0.3-4.4), and 0.5 (0.1-2.8) mumol.L-1, respectively. Apa did not inhibit NO production. CONCLUSION: GA and GB inhibited NO production in LPS-stimulated microglia. PMID- 10375813 TI - Nitric oxide and soman poisoning. AB - AIM: To examine whether nitric-oxide (NO) is involved in the toxicity of soman. METHODS: With pretreatments of icv L-arginine (Arg, the substrate of nitric-oxide synthase NOS), NG-nitro-L-arginine methyl ester (NAME, the inhibitor of NOS), the latency of seizure, and the mortality of mice induced by soman poisoning were examined. The activities of brain NOS in soman-intoxicated mice were measured. RESULTS: In case of Arg pretreatments, the latency decreased (P < 0.05) from (5.2 +/- 1.8) min (control) to (4.3 +/- 0.8) min (Arg 160 nmol), and the mortality increased (P < 0.05) from 50% (control) to 81% (Arg 160 nmol). In case of NAME pretreatment, the latency increased (P < 0.01) from (4.0 +/- 1.1) min (control) to (14.5 +/- 5.0) min (NAME 2.20 mumol), and the mortality decreased (P < 0.05) from 87% (control) to 50% (NAME 2.20 mumol). The toxicity of soman in mice was enhanced by Arg and reduced by NAME all in a dose-dependent fashion. NAME antagonized the enhancement of soman poisoning by Arg. Intoxication of mice with soman increased the NOS activity in cerebrum, cerebellum, and hippocampus from 100% to 104% (P < 0.05), 115% (P < 0.01), and 111% (P < 0.01), respectively. CONCLUSION: The onset of seizure and death of mice induced by soman poisoning are related to the NO messenger system. PMID- 10375814 TI - Tyrosine kinases participate in alpha 1A-adrenoceptor-mediated vasoconstriction in perfused rat hindlimb. AB - AIM: To determine whether or not tyrosine kinase is involved in the signal transduction of alpha 1A-adrenoceptors. METHODS: Effects of various pharmacological probes on norepinephrine (NE)-induced vasopressor responses were determined in the perfused rat hindlimb. RESULTS: The putative tyrosine kinase inhibitors, genistein, and tyrphostin, significantly inhibited the vasopressor responses induced by NE but not that induced by KCl. The protein-tyrosine phosphatase inhibitor, sodium orthovanadate, selectively potentiated the vasopressor responses induced by NE. Neither genistein nor tyrphostin had effect on the contraction elicited by phorbol 12-myristate 13-acetate. In contrast, both genistein and tyrphostin attenuated the vasopressor responses evoked by NaF. CONCLUSION: The genistein- and tyrphostin-sensitive tyrosine kinases participate in alpha 1A-adrenoceptor-mediated vasoconstriction in perfused rat hindlimb. PMID- 10375816 TI - Characteristics of transient outward K+ current in human atrial cardiomyocytes. AB - AIM: To study the properties of transient outward potassium current (Ito) in human atrial myocytes. METHODS: The patch-clamp whole-cell recording techniques were used. When the calcium inward current was blocked by CdCl2 (0.1 mmol.L-1), the transient outward potassium current (Ito) was recorded by depolarizing cells. RESULTS: The current was voltage dependent, it was activated quickly and inactivated rapidly too. 4-Aminopyridine (4-AP) 10 mmol.L-1 (a selective blocker of Ito) blocked the current completely. The IC50 (95% confidence limits) of 4-AP on Ito were 0.67 (0.58-0.80) mmol.L-1, 4-AP 1 mmol.L-1 shifted the activation curve of Ito to positive potential, therefore a higher membrane potential was required to activate Ito. CONCLUSION: Ito, a major K+ channel current in human atrial cells, is blocked by 4-AP. PMID- 10375815 TI - Mediation of calcitonin gene-related peptide in protection of ischemic preconditioning in rat hindlimbs. AB - AIM: To study modulation of calcitonin gene-related peptide (CGRP) in the protective effect of ischemic preconditioning on endothelial cells. METHODS: Rat hindlmbs were subjected to ischemia for 2 h, and endothelium-dependent vasorelaxation to acetylcholine (ACh) was examined in rat hindlimbs. RESULTS: Two hours of ischemia elicited no effect on vasoconstrictor responses to norepinephrine, but markedly impaired vasodilator responses to ACh. Ischemic preconditioning induced by 5-min aortic occlusion and 10-min blood reperfusion prevented the impairment of vasorelaxation to ACh due to long-term ischemia. The protection of ischemic preconditioning was abolished by repeated pretreatments with capsaicin to deplete CGRP. Acute application of capsaicin to evoke CGRP release or CGRP caused an ischemic preconditioning-like protection. CONCLUSION: Capsaicin-sensitive sensory nerves are involved in the protective effect of ischemic preconditioning on endothelial cells in the rat hindlimbs, and CGRP can mimic the protective effect of ischemic preconditioning in blood vessels. PMID- 10375817 TI - Inhibitory effects of S-nitrosocaptopril on vasomotor tone. AB - AIM: To study effects of captopril (Cap) and S-nitrosocaptopril (CapNO) on vascular tension. METHODS: Tension of rabbit aortic rings and perfusion pressure of rat renal artery (PPr) were examined to evaluate the effects of CapNO. RESULTS: CapNO (3 nmol.L-1-10 mumol.L-1) produced concentration-dependent vasorelaxation response in the rings of rabbit thoracic aortas. Endothelium denudation did not alter the relaxations to CapNO. In contrast, Cap had no vasorelaxing effect on the rings precontracted with phenylephrine. CapNO (10 nmol.L-1) decreased rat PPr in vivo (P < 0.01), and the effect was concentration dependent and reversible. Cap showed a reduction in rat PPr only at the concentration 1000 nmol.L-1 (P < 0.05). The relaxing potency of CapNO was 100 times higher than that of Cap in this respect. Pre-perfusion of rat renal arteries with NG-monomethyl-L-arginine monoacetate (L-NMMA, 0.03 nmol.L-1) or L arginine (3 nmol.L-1) did not significantly blocked the relaxing effect induced by CapNO. CONCLUSION: CapNO had a direct vasodilatory effect. PMID- 10375818 TI - Effect of antisense mitogen-activated protein kinase oligonucleotides on rat vascular smooth muscle cell proliferation induced by EGF in vitro. AB - AIM: To study the preventive effect of down-regulating mitogen-activated protein kinase (MAPK) on vascular smooth muscle cell (VSMC) proliferation. METHODS: Cultured rat VSMC was pretreated with a phosphorothioate-protected 17-mer antisense MAPK oligodeoxynucleotides (ODN) directed against the initiation of translation sites of the p42- and p44-MAPK isoforms by liposomal transfection. A 17-mer sense and a random sequence MAPK ODN were used as control. After liposomal transfection, cells were exposed to epidermal growth factor (EGF) 1 nmol.L-1 for 10 min and then harvested in lysis buffer. MAPK activity was measured by Western blot and P-81 phosphocellulose filter papers method by using [gamma-32P] ATP and myelin basic protein as substrate. DNA synthesis was measured by [3H]thymidine incorporation. RESULTS: Antisense ODN 0.2 mumol.L-1 reduced EGF-induced MAPK activities by 84%, and inhibited VSMC [3H]thymidine incorporation stimulated by EGF. CONCLUSION: A 17-mer MAPK antisense oligonucleotide directed against the initiation of translation sites of the p44- and p42-MAPK inhibited EGF-stimulated rat VSMC proliferation. PMID- 10375819 TI - Effect of histidine on myocardial mitochondria and platelet aggregation during thrombotic cerebral ischemia in rats. AB - AIM: To study the effect of histidine on cerebral thrombosis and possible mechanism. METHODS: Cerebral-cardiac stroke was produced by photochemically induced thrombotic cerebral ischemia in rats. RESULTS: Platelet aggregation in whole blood increased markedly, peak heights at 4 and 24 h were (5.1 +/- 0.5) omega and (4.3 +/- 0.5) omega, respectively. Heart mitochondria volume (V), volume density (Vv), surface density (Nm), and surface density of outer membrane (Sv1) increased (8.2 +/- 5.5, 0.59 +/- 0.16, 0.11 +/- 0.03, and 0.22 +/- 0.05, respectively, P < 0.01), but numerical density (Nv), specific surface of inner membrane (delta 2) and of the cristae (delta 3) decreased (0.07 +/- 0.02, 2.8 +/- 0.8, and 2.4 +/- 0.7, respectively, P < 0.01) after cerebral thrombosis. The myocardial histopathologic characteristics were different from those of ischemic necrosis and myocardial damage caused by ischemic reperfusion. In rat treated with histidine after photochemical reaction, platelet aggregation decreased markedly [(2.93 +/- 1.08) omega, P < 0.01], reversible change often went with parameters related to the inner mitochondrial membrane but not the outer mitochondrial membrane. CONCLUSION: Histidine depressed platelet aggregation and reduced myocardial mitochondrial damage resulted from cerebral ischemia. PMID- 10375820 TI - [Effects of O-(4-ethoxyl-butyl)-berbamine on isolated frog hearts]. AB - AIM: To study the effects of O-(4-ethoxyl-butyl)-berbamine (EBB) on isolated frog heart. METHODS: The isolated frog hearts were prepared with Straub's method; heart contractions were recorded using LMS-2A recorder by tonotransducer. RESULTS: EBB 1-100 mumol.L-1 caused concentration-dependent increase in systolic and diastolic activities, but did not alter the heart rate. EBB 250-500 mumol.L-1 slowed the heart beats and caused ventricular asystole. Cimetidine 10 mumol.L-1 or prazosin 100 mumol.L-1 did not inhibit the positive inotropic action of EBB. Whereas verapamil 0.01 mumol.L-1 antagonized the positive inotropic action of EBB. EBB enhanced positive inotropic action of CaCl2. The action of EBB was slower but longer than caffeine or isoproterenol. At room temperature (21.0 +/- 0.5) degree C, however, the onset of oubain was slower, but the duration of its peak action was longer, than EBB. CONCLUSION: The positive inotropic action of EBB was, at least partially, via promoting the inward current of [Ca2+]o, rather than increasing the intracellular Ca2+ release. PMID- 10375821 TI - [Study on the analysis of methamphetamine and its metabolite amphetamine in blood and urine]. AB - The methods using GC/MS,GC/FID,GC/NPD for identification and quantification of Methamphetamined(MAMP) and Amphetamine (AMP) in human blood and urine were developed. Using 4-phenylbutylamine as the internal standard, the samples were extracted with 200 microliters cyclohexane and then the 2 microliters cyclohexane was injected into GC or the extracts were derivaized by using microwave irradiation. The methods given allowed simple and rapid procedure, the recoveries were greater than 80% and the sensitivity limits were 2-5 ng/ml. The methods for d/L Enantiomer of MAMP and AMP in human urine was also described, which can be used to determine the source of Amphetamines and determine the toxic effect in the case. PMID- 10375822 TI - [Experimental study of myocardial myoglobin depletion induced by early myocardial infarction and coronary artery spasm]. AB - In this study, the model of early myocardial infarction (EMI) was produced by legation of left anterior descending artery in rat. The artery spasm (CAS) was produced by external jugular vein injection of vasopressin (VP). Myocardium of apex and adjoining slice was used to make paraffin sections, then HE and LSAB-Mb staining were performed. Results showed: In 25 min after myocardial infarction, stretch-shape Mb depletion was observed in subendocardial myocardium of aortic ventricle's frontal wall. As the time of EMI prolonged, stretch-shape Mb depletion extended to epicardial layer. In CAS group, multiply, spotty Mb depletion was detected. There were more Mb depletion zones in right heart than that in left heart. The Mb depletion zones surrounded the CA or were like grape clusters along the branches of one large CA. Thus, myocardial Mb depletion induced by EMI and CAS had different morphologic peculiarity. LSAB-Mb method could be hoped for the use of supplying objectively morphologic evidence to myocardial ischemia induced by CAS. PMID- 10375823 TI - [Immunohistochemical study on SMDS using anti-actin monoclonal antibody]. AB - For elucidating the relationship between the causes of sudden manhood death syndrome (SMDS) and cardiac sudden death, the cardiac conducting system-(CCS) and myocardium of 23 cases including SMDS group, coronary heart disease group and normal control group were studied using anti-actin monoclonal antibody (HHF35) S P immunohistochemical method. The results showed that some areas of CCS and myocardium were stained negatively by HHF35 in SMDS group but no infarction area can be found with H.E staining. The negative staining by HHF35 can be also observed in all cases of coronary heart disease group. In control group, the CCS and myocardium stained positively. It indicated that there were pathological changes such as early ischemia or infarction of myocardium in SMDS, which related closely to cardiac sudden death. PMID- 10375824 TI - [Study on cerebral ATP level with bioluminescent: method to estimate the time of death]. AB - This paper reported the changes of ATP content in dog's cerebral tissues during different time after death. ATP levels were determined by biolumi-nescent method. The results showed that ATP concentration decreased gradually along with prolonging of death time. The study has important value in estimating the time of death in forensic practice. PMID- 10375825 TI - [Observation on postmortem tracheal cilia of drowning by scanning electron microscope (SEM)]. AB - The changes of tracheal cilia of drowning rat were observed by SEM at 0, 24, 48, 72, 96 h after death. The authors found: the tracheal cilia lost their normal arrangement in some area, and the opening increased on the top of the Goblet cell. There were some thing resembling cotton, or looking like network on the surface of tracheal cilia. At different times after death, the tracheal cilia appeared adhering, downing, and destroying, Goblet cell membrane creasing, the top of cell flattening and the cell collapsing. Observing this changes, we could estimate the time of death. PMID- 10375826 TI - [Legal bases for evaluation of mental injuries]. AB - In this article, we study the legislation bases for evaluation of mental injuries, the legislation form of the evaluation standard, and also discuss the degrees of mental injuries. PMID- 10375827 TI - [Analyses of 430 cases of limb injuries]. PMID- 10375828 TI - [Analyses of expert's testimonies about 22 mental patients occurred in trip]. AB - This article reviewed the expert's testimonies about 22 mental patients involving in cases of murder and harm. Of these patients, 20 are male, 2 female. Their mean age is 31 +/- 9.72% of them are peasant and non-regular workers. The results showed that all actions of murder and harm were induced by the direct effect of auditory hallucination and disturbance of consciousness, lacking premediated murder and self-protection. The people attacked are mostly strangers and passengers. All of these patients were considered non-responsibility and non ability legally. The problems about distinguishing features and clinical diagnosis of the diseases were discussed. PMID- 10375829 TI - [Typing of ABH blood group on human nails by improved two-dimensional absorption method]. AB - The two-dimensional absorption-inhibition method has been proved to be more sensitive than any other classical method in detecting ADH antigens in human bone, dental and urine tissue. The 96F microwell plates are used in our experiment, for it is easy for us to observe the results. It shows that the Two dimensional absorption-inhibition method is practically effective to typing of ABH antigen on human nails. PMID- 10375830 TI - [Gunshot residue particle detected in entrance wound of near range fire with energy dispersive X-ray]. PMID- 10375831 TI - [The use of microwave for immunohistochemical technology in forensic pathology]. AB - A microwave oven (MWO) of National brand was applied for Labeled Streptavidin Biotin immunohistochemical staining (LSAB method) on autopsy materials of 20 cases to detect 10 kinds of antigen of heart and brain: The keys to manipulate MWO as follows: (1) Recovery of antigen: submerge sections in 0.01M citrate buffer (pH 6.0), turn MWO to the position of DEF (210W) for 10 min, reaching temperature 92 degrees C-98 degrees C. (2) add normal serum, first antibody, second biotinylated antibody and streptavidin complex, then turn MWO to the position of LOW (70W) for 5 min, reaching temperature 20 degrees C-37 degrees C. Other steps were the same as the standards. Results showed: MWO can recover the destroyed or covered antigen, improve positive staining degree, reduce background's stain and shorten staining time. PMID- 10375832 TI - [Macro, micro and ultrastructural observations of pituitary, adrenal, pancreas and spleen in experimental brain death]. AB - The purpose of this study is to observe the macro, micro and ultrastructural changes of the pituitary, adrenal, pancreas and spleen in the experimental primary brain death. The results showed: 1. acinar zymogen granule of pancreas decreased, karyopyknosis of islet cells formed. 2. dilation and blood stasis of spleen sinus happened 3. adrenocortical karyopyknosis formed, mitochondrial crista vanish. 4. Pituitary karyopyknosis and cytolysis developed. 5. there was correlation between the extent of lesions and time after brain death. PMID- 10375833 TI - [The identification of banding chromosomes by computer image analysis]. AB - The analysis system for banding chromosomes (SYSTEM), an image analysis software, was programmed. By the SYSTEM the photos of Chinese G banding chromosomes were analyzed. It showed that the karyotype of the chromosomes of persons was recorded in high speed and efficiency, and that the chromosomes of alike ordinal number from different persons could be compared directly. That would presage, by this kind of the high effective operation, a favorable condition was created for the application of the polymorphism of the chromosomes to the parentage diagnosis. PMID- 10375834 TI - [The application of microwave irradiation to derivatization]. AB - The methods using microwave irradiation for rapid preparation of acetyl, trifluoroacetyl, pentafluoropyl, heptafluorobutyl and MSTFA derivatives of Morphine, AMP and MAMP were developed. Conventional techniques for the reaction mixture to be heated need 30 min at 60-70 degrees C to form derivatizations, but by microwave irradiation, only 2-3 min is needed. The mass spectral fragmentation patterns and gas chromatographic retention times of the derivatives obtained by both microwave irradiation and conventional heating are similar. PMID- 10375836 TI - [Studies on basis and methods for examination of ruled characters]. AB - Although the writer of ruled characters temporarily changes his movement habits forcefully, the static impression of characters in mind still determines the characteristics of structures and wrongly written. The static impression controls ruled writing. This builds up the basis for the examination of ruled characters. The main methods for examination is to find in characters special structures, stroke collocation, and wrongly written. PMID- 10375835 TI - [Determination of basic drugs in blood by RP-HPLC]. AB - A reversed-phase HPLC method for determination of phenothiazines and tricyclin antidepressants in whole blood was described. In this paper, a model 1090 HPLC with DAD and a zorbax ODS column was used. The mobile phase was methanol: water: tritely amine (75:24.7:0.3) with pH 7.5. Cyproheptadine was used as internal standard in this method. Blood samples were extracted with the solid-phase extraction method and the liquid-liquid method. This method is suitable in forensic toxicology analysis for basic drugs. PMID- 10375837 TI - [An autopsy analysis on 14 cases of poisoning death caused by illegal quackery]. AB - In this paper, 14 cases on poisoning death caused by illegal quackery were analyzed. The results indicated that the causes and routes of poisoning were various, and that there were three kinds of poisoning types in illegal quackery. Main points of forensic identification were pointed out. In addition, some medical and administrative measures and possible measures were also investigated. The authors suggest that measures as above are of great importance for preventing similar poisoning and improving forensic identification. PMID- 10375838 TI - [Analysis of 36 cases of traumatic cerebral infarction]. AB - 36 cases of traumatic cerebral infarction were analysed. The clinical signs and symptoms, pathological mechanism, relation between injury and disease and degree of injury were discussed. PMID- 10375839 TI - [Analysis of 69 cases of pudendal injuries]. AB - Cause of trauma, tools which cause injuries and types of injuries were analyzed in 69 cases with pudendal injuries. The direct outcome of the injury were discussed. PMID- 10375840 TI - [Advances in the study on immunoassay of narcotic drugs]. PMID- 10375841 TI - How to use low-molecular weight heparin for outpatient management of deep vein thrombosis. AB - Low-molecular weight heparins can be used to treat acute deep vein thrombosis on an outpatient basis, but such use requires careful planning and patient education. We present an algorithm used at the Cleveland Clinic. PMID- 10375842 TI - A 57-year-old woman with pulmonary infiltrates and eosinophilia. PMID- 10375843 TI - A common-sense approach to chronic fatigue in primary care. AB - Since chronic fatigue is so common, long-lasting, and fraught with functional and emotional consequences, early intervention can limit overuse of health care resources and forestall disability. To help patients, we must intervene before chronicity is established or iatrogenic harm has occurred. Care that integrates medical and psychologic concepts, together with symptom management, can prevent significant secondary impairment in most cases. PMID- 10375844 TI - Sones' laws of clinical conduct. PMID- 10375845 TI - Review of newer contraceptive agents. AB - Advances in contraceptive technology have made birth control more effective, convenient, and safe. We review the newer products and some under development, including the latest oral contraceptives, injectable progesterone, subdermal progestin implants, progesterone-releasing IUDs, emergency contraception, and male contraception. PMID- 10375846 TI - Inhibitors of tumor necrosis factor: new treatment options for rheumatoid arthritis. AB - Infliximab and etanercept, both approved by the FDA in 1998, are examples of a new class of disease-modifying antirheumatic drugs that interfere with the action of tumor necrosis factor alpha, one of the key cytokines that promote inflammation. Infliximab is approved for Crohn disease and etanercept for rheumatoid arthritis. Both show promise in treating rheumatoid arthritis, although the long-term risks and benefits of these drugs are not yet known. PMID- 10375847 TI - Listeriosis: recognizing it, treating it, preventing it. AB - Listeria monocytogenes has become a major pathogen in foodborne illness. It most often affects patients who are pregnant, at the extremes of life, or immunocompromised in some way. A variety of clinical manifestations are possible, but bacteremia and meningitis are most common. This article reviews the epidemiology, microbiology, populations at risk, clinical manifestations, treatment, and prevention of listeriosis. PMID- 10375848 TI - [Study of the permeability of oral leukoplakia]. AB - The significant increase in the mortality rates of oral carcinoma speeded up the research on the mechanisms of tumor promoting and inhibiting factors. Results of previous studies in the USA showed different permeability between keratinized (hard palate, gingive) and non-keratinized (floor of the mouth) regions of the oral mucosa, and a corresponding difference in lipid composition. Aim of this study was, to compare the permeability and lipid composition of leukoplakia of the oral mucosa with healthy specimens from the same region, in order to assess a possible enhancement or inhibition in the barrier-function, in cases of pathological keratinization. Results of the measurements showed, that hyperplastic leukoplakia areas were more permeable for the NNN carcinogens contained in tobacco, and even clinically healthy sites showed a higher permeability, that oral mucosa of non-smokers. These data together with the results of lipid analysis, point to the possible generalized changes--caused by smoking--in the oral cavity, even before the appearance of clinical signs. PMID- 10375849 TI - [The development and action-mechanism of the quadhelix in orthodontic treatment]. AB - Authors present a historical review of the development of quadhelix appliance. In conclusion, slow maxillary expansion with the quadhelix appliance provides an alternative approach to rapid maxillary expansion that is particularly beneficial for correction of transverse and arch-length discrepancies during the mixed dentition. There are numerous indications for this approach with two major advantages. First, a lower force that is within the elastic limits of maxillary sutural and periodontal tissue is generated. Second, a low separation rate of 0.4 1.1 mm per week allows for more physiologic adaptation to the expansion that may result in a more stable correction with less sutural relapse. The quadhelix is a versatile appliance that provides practitioners a convenient means to normalize developing malocclusions. PMID- 10375850 TI - [Comparison of the effect of acetone-containing and acetone-less bonding materials on dental pulp blood vessels in rats]. AB - No data are available on the direct acute vascular effect of dental bond materials in the dental pulp. The purpose of the present study was to investigate the effect of composite resin bonding systems on the pulpal vascular diameter. Three groups of male Sprague-Dawley rats (weighing 300-490 g), each of 10 animals were used for this investigation. The left lower incisor of the rats was prepared for vitalmicroscopy. Changes in vessel diameter were recorded prior to; and 5, 15, 30, 60 minutes after the application of saline (control) or the bonding agents (test1: acetone containing--/Prime/Bond2.1, DeTrey/; test2: acetone free /Scotchbond Multi-Purpose Adhesive System, 3M/ bond material) on pulpal dentin as recommended by the manufacturer. In control rats, the vessel diameter was stable during the experiment. However, in the presence of bonding materials an enhancement was registered in vascular diameter (p < 0.05). The bond materials applied directly onto a very thin layer of dentin show acute vasodilating effect on the rat pulpal microvessels, but no stasis or prestasis has been detected, indicating a possible reversible effect. We could not show any statistical difference between the vasodilatation caused by the acetone containing and the acetone free bonding material. PMID- 10375851 TI - [Epidemiologic survey of tongue lesions and analysis of the etiologic factors involved]. AB - The prevalence of tongue lesions, and relationships with different systemic diseases, according to the international literature of epidemiologic studies were reviewed. The data were compared with a Hungarian investigation of 5034 individuals, between 1992 and 1995 in Budapest. Tongue lesions were found in 22.76% of the examined individuals. The ratio was similar in women (51%) and men (49%). Fissured tongue was found in 21.49%, geographic tongue in 2.21%, atrophic tongue in 0.12% and central papillar atrophy in 0.70%. These data agree in general with the epidemiological data found in the international literature. Some rare tongue alterations were also investigated. Lingua indentata was found in 1.49% and hypertrophy of papillae foliatae was found in 0.20%. The relationship with different systemic diseases was the following: In diabetes mellitus tongue lesions were found in 29.03%, in hypertension in 28.63%, connected to heart- and vascular diseases in 25.15%, to haematologic diseases in 17.54%. Tongue lesions were found in 23.86% in the case of liver disease, in 22.38% in gastrointestinal diseases and in 20.69% associated to tumors. In the case of Candida's infection tongue alterations were found in 41.6%, in smokers in 23.72%. Tongue lesions caused complaints rarely, only in 2.27%. The recognition of tongue lesions may be helpful in the early diagnosis of systemic diseases. PMID- 10375852 TI - [The sequence analysis of the 3' end of intron 15 from human LDL receptor gene and its application in the study of gene diagnosis for familial hypercholesterolaemia]. AB - To understand intron 15 of human LDL receptor gene, the DNA fragments from exon 15 to exon 16 and the 3' end of intron 15 were amplified with long chain PCR and anchored PCR. The 3' end of intron 15 was sequenced with Dynalbeads-Streptavidin Solid Phase technique. The sequence analysis showed that the 3' end of intron 15 contained the 3' splicing site and the branch site at 31 nucleotides upstream of the 3' end. Besides the authentic branch site, it is possible that the 3' end of intron 15 contains a cryptic site (GCCTCAC) at 20 nucleotides upstream of the 3' end. The sequences suggest that the PvuII polymorphism at Intron 15 is caused by the T-C substitution. According to the sequences of the 3' end, the new PCR-RFLP protocol for detection of PvuII polymorphism at intron 15 was developed. Using this protocol a representative familial hypercholesterolaemia family was identified with linkage analysis of PvuII polymorphism at intron 15. PMID- 10375853 TI - [Gene cloning and expression of mouse type I soluble interleukin 1 receptor in insect cells]. AB - Interleukin 1(IL-1), which takes part in many physiological and pathological processes, is a kind of important cytokines. Their effects are exerted via specific IL-1 receptors (IL-1R) which are present on a wide variety of different cell types. Two types of IL-1R are identified as type I receptor (IL-1RI) and type II receptor (IL-1RII). Soluble forms of IL-1R can be produced from proteolytic cleavage of the extracellular portion of the two type receptors on cell surface. In this study, mouse type I soluble IL-1R (sIL-1RI) has been cloned and expressed in insect cells Sf9 with baculovirus as vector. Total RNA from NIH/3T3 cell was extracted with guanidinium thiocyanate followed by ultra centrifugation in cesium chloride solution. The cDNA of sIL-1RI was amplified by RT-PCR technique. The cDNA was cloned into plasmid pAcGP67B. The recombinant plasmid pAI (pAcGP67B-sIL-1RI) was cotransfected with wild type AcNPV DNA into insect cells Sf9, because of genetic exchange occurred by homologous recombination in vivo, recombinant baculovirus rAcNPV was produced. Insect cells Sf9 were infected with the purified rAcNPV and sIL-1RI gene were expressed. SDS PAGE analyses and Blockage assay of IL-1 beta biological activity demonstrated that the recombinant sIL-1RI had biological function and could be secreted into the medium. PMID- 10375854 TI - [A cis-regulatory enhancer element of the nodal gene is present in its first intron]. AB - The mouse nodal gene is expressed during early embryogenesis and plays an important role during gastrulation and the determination of the left-right body symmetry/axis formation. The embryonic stem (ES) cell line 413-d 3563 contains a single copy of a retrovirus insertion in the first inton of the nodal gene. Using a quantitative RNase protection assay, the level of the nodal gene expression in this cell line was compared to that of the parental CCE cell line. A 2.3 fold down regulation of the nodal transcript in the 3563 ES cell line suggested that the proviral insertion might decrease the nodal mRNA expression by disrupting the potential cis regulatory elements present at the insertion site in the first intron of the nodal gene. Luciferase reporter constructs containing the entire first intron fragment or part of it were tested by a transient transfection assay for their potential transcriptional regulatory activity. A 2.4 kb DNA fragment from the 5' end of the intron was sufficient to increase the expression level of the reporter gene 8 fold, suggesting that an important cis acting enhancer element for the nodal gene expression is localized within the first intron. PMID- 10375855 TI - [The influence of the 20 kDa protein from Bacillus thuringiensis subsp. israelensis on the cytolytic activity of CytA]. AB - In order to understand the influence of the 20 kDa protein on the cytolytic activity of CytA protein, a set of PCR primers were designed to amplify 20 kDa protein and CytA protein genes. The genes were ligated to E. coli expression vector pUHE24 and transformed into E. coli XL1 and DH5 alpha. The clones, LZ20, LZcytA and LZ20A containing 20 kDa protein gene, cytA protein gene and 20 kDa cytA genes were obtained respectively. The influence of the clones on the growth of E. coli cells was determined under induction of IPTG. The results showed that the growth of LZ20 cells were not affected, LZcytA cells were killed, and the growth of LZ20A cells were not affected. It was suggested that expression product of 20 kDa protein gene protected the host cells from the cytolytic effect of CytA protein. This was supported by using different host strains. PMID- 10375856 TI - [Expression of chimeric single-chain antibody with specificity for HBsAg in E. coli]. AB - The Vk gene of MAb with specificity for HBsAg and human Ck gene have been combined to form chimeric light chain (Vk-Ck) by recombinant PCR, which has combined with VH to construct chimeric single-chain antibody(ScFv-Ck) by a linker encoding a flexible peptide[(Gly3 Ser)3]. The ScFv-Ck has been expressed in the heat-induced expression system and secreted expression system of E. coli respectively. Analysis by Western-blot and indirect ELISA shows that the ScFv-Ck product in two systems both have HBsAg-binding ability, and ScFv-Ck has been secreted from E. coli in the direction of secretion peptide. PMID- 10375857 TI - [Relationship between the familial nonrandom chromosome loss (NCL) and leukomogenesis]. AB - By using R-banding karyotypic analysis technique, the bone marrow (BM) cells were performed in 223 hematopoietic malignacies and 105 diopathic throbocytopenic purpura (ITP), which served as control. The following results were obtained: (1) Nonrandom chromosome loss (NCL), such as, -11, -14, -21, etc, which were found in the affected members of leukemia families, were found in about 30% sporadic ANLL, MDS and about 50% ALL, espedislly in 100% (5/5) CLL, but not found in ITP (P < 0.001). These results indicated that the familial nonrandom chromosome loss were associated with leukomogenesis. (2) Because most of BM cells are hypodiploed and have the same kinds of NCL in each cases of CLL, which can develop into ALL, ANLL and also cancers, ALL BM hypo- and hyper-diploid and/or polyploid cells might be origin of hypodiploid cells. (3) 28% (6/21) of pediatric patients with AL, MDS, or FA (Fanconi Anemia) have one parent, who have up to 30% BM hypodiploid cells and similar kinds of NCL and also have the rearrangement of C-erbB and abnormal proliferation of BM. The NCL were found in the three consecutive generations of a family with 5 ALL among 10 members of third generation. It indicated that the familial NCL might be inherited and be coded by a unknown gene alteration, which might be related to leukomo and carcinogenesis, because there are genes or their candidates for leukemia in the chromosomes 11, 14 and 21. (4) Based on the works of my colleages and I, the model of leukomo and carcinogenesis was proposed and the relationship between chromosome monosomy, deletion, translocations and leukomogenesis were showed elswhere. The significance of monosomy 11, 14 and 21 etc. were discussed briefly. PMID- 10375859 TI - [Molecular mechanism of diapause in Bombyx mori. 2. Diapause determination and expression of diapause hormone gene in pupal stage]. AB - Embryonic diapause in Bombyx is induced by diapause hormone (DH), which is produced in the suboesophageal ganglion from a precursor protein. We have cloned the DH cDNA into pBluescript KS vector that containing T7 RNA promoter can be used to synthesize nonspecific RNA. DH cRNA was synthesized and used as internal control. We quantitatively measured DH mRNA at SG of day 3 pupae by Northern hybridization analysis and suggested that the embryonic diapause of Bombyx in next generation was determined by DH mRNA content in SG. PMID- 10375858 TI - [Construction of expressing vector for phage display scFv and a mouse unspecified antibody library]. AB - The display of antibody gene library on the surface of E. coli filamentous phage offers a new method of obtaining antibodies against specific antigens. In the paper, it is reported that a phagemid for phage display antibody, named after pFUW80, characterized expressing scFv either secretly or stickily. With a series of designed PCR primers, heavy-chain and light-chain variable region genes of mice antibodies were amplified and a mouse unspecified single-chain antibody library with the size of 1.2 x 10(6) clones was constructed. From this library, phage was selected out against antigen human IgG and detected by ELISA and partial sequence analysis. These primary results established the foundation for future research using this system. PMID- 10375860 TI - [Aneuploidy detection by multi-color fluorescence in situ hybridization in mouse sperm]. AB - Multi-color fluorescence in situ hybridization (FISH) was employed to evaluate the frequency of aneuploid sperm in young adult mice after treated by 2-(4' thiazolyl) benzimidazole or thiabendazole (TB). The animals were given TB (100 200 300 mg/kg) by oral incubation daily for 11 days, and sacrificed 22 days after the last treatment of TB. Three-chromosome FISH was applied to determine the frequencies of diploid, disomic and nullisomic sperm with DNA probes specific for the chromosomes X, Y and 8. The aneuploid frequencies were not significantly different between the sperm in 100 mg/kg group and solvent control. The frequencies aneuploid and diploid sperm in middle dose group were significantly higher than the solvent control (P < 0.001-0.05). There were no significant aneuploid frequency increases were found in 300 mg/kg group excepting the frequency of diploid sperm. It is conclude that TB is a potential germinal aneugen in mammalian, which may block the cleavage during the meiosis. Epididymal sperm aneuploidy can be reliably determined by sperm FISH analysis in mice. PMID- 10375861 TI - [Formation of germline chimeras from murine embryonic stem cell lines]. AB - Generation of germline chimeras is the crucial step in ES cell-mediated transgenesis. The prerequisite for germline chimerism is the maintenance of germline differentiating potency of ES cells, whereas production of germline chimeras is the only method to prove whether such potency is maintained. In order to investigate the germline differentiating potency of three newly established ES cell lines (MESPU21, MESPU22 and MESPU29), ES cells were introduced into host embryos from inbred C57BL/6J and outbred KMW or ICR through blastocyst injection or 8-cell stage morula injection. Totally 81 chimeras were obtained; among 42 test-bred ones, 19 were germline transmitters assessed by coat analysis, as is the first report of ES cell-embryo germline chimeras in China. MESPU21 and MESPU22 formed germline chimeras in high frequency and most of those chimeras produced ES cell-derived progeny in high proportion, which proved that both ES cell lines retained good germline differentiating potency and could be used as valuable cellular vehicles to introduce genetic modifications into mouse genome. PMID- 10375862 TI - [Genetics and expression stability of exogenous gene construct in transgenic mice]. AB - Transgenic mice were produced by introduced exogene construct lambda 106, an expression construct of HBsAg gene directed by bovine alpha-S1 casein gene, with microinjection. Gene integration test with PCR-Southern hybridization, shows that the construct integration rate is 56% (17/30) and expressed rate, by ELISA, of target gene product HBsAg is 100% (8/8) in the first generation. Generation trailing test suggests that the gene construct can be stably inherited across generation after generation, and expressed more or less than parents among offspring. However positive rate of offspring does not follow genetic ratio that exogenous gene can be integrated randomly into a single site on chromosomes. There may be a machinery of multi-sites for exo-gene integration. Changes of the target gene expression in the offsprings may be related to position effect, changing of gene copy number and changing of chromatin imprinting pattern when parent's genome information was delivered to their progeny. PMID- 10375863 TI - [Delineation of the cis-regulatory sequences of the nodal gene]. AB - In mouse embryo development, nodal is expressed in the node during the gastrulation. It is involved in mesoderm formation and left-right axis determination. The expression of the nodal gene has a restricted spatial and temporal specificity. Studies on the function of the nodal first intron in the gene expression regulation have shown that a 220 bp fragment at the 5' end of the first intron has an enhancer activity. Rather unexpectedly, the transient transfection assays in F9 cells indicate that this fragment also displayed a promoter activity when it is cloned upstream of basic luciferase reporter gene only in sense orientation. We present here the nucleotide sequence of this 220 bp fragment, and discuss its possible role in the nodal gene regulation. PMID- 10375864 TI - [Immunolocalization of actin in synaptonemal complexes in spermatocytes of Mesocricetus auratus]. AB - Opinions upon whether actin is present in synaptonemal complexes of eukaryotes are still controversial. We detected actin in synaptonemal complexes of spermatocytes of Mesocricetus auratus with indirect immunofluorescence technique and observed bright fluorescence in the SCs, indicating the presence of actin in the structure. We labelled sections of the spermatocytes of M. auratus with immunogold technique, gold particels were observed over the lateral elements and attachment plaques of the SCs, and many of the gold were found in clusters, confirming the results of the immunofluorescence observations. Further examinations revealed the gold particles representing actin in SCs of autosomes and sex chromosomes and also in SCs of zygotene and pachytene. Our results are in favour of the point of view that actin is a component of the SC. PMID- 10375865 TI - [A study of a derivative of Taq DNA polymerase]. AB - Using the method of double primer oligonucleotide-mediated mutagenesis, the high expression plasmid of TaqND236, a derivative of Taq DNA polymerase, was constructed. To determine the frameshift mutation frequency of the in vitro DNA synthesis, we constructed a Gapped-DNA system using the pFDPM118 (a mutant of pUC118 with a -1 frameshift mutation on the lacZ gene) as template. By calculating the ratio of blue and white colonies on the X-gal plate after transforming E. coli TG1, the frameshift mutation frequency of Taq and TaqND236 was measured. It was found that the replication fidelity of the deleted Taq TaqND236 increased more than 10 folds. PMID- 10375866 TI - Characteristics of primary osteoblast culture derived from rat fetal calvaria. AB - Primary osteoblast cultures, which reflect more phenotypic properties of normal osteoblasts than osteoblastic cell lines, can be used as an experimental tool for investigating the osteoblastic functions in vitro. Primary osteoblast cultures were obtained from the parietal bones of calvaria of fetal rats in this study. Differential characteristics of osteoblasts in our culture system were examined and fibroblast cultures were also tested for comparison. We tested the alkaline phosphatase (ALP) and von Kossa stains on osteoblast and fibroblast cultures to examine the expression of ALP and the subsequent matrix mineralization occurred at 2 and 3 weeks after cell confluence respectively. The results showed that osteoblast cultures revealed obvious positive stains of ALP and von Kossa, while fibroblast cultures revealed negative stains, suggesting the osteoblast culture system used in this study reflects the typical phenotypes of primary osteoblasts but not fibroblasts. We tested the ALP activities following various doses of PGE2 or ketorolac treatments in primary osteoblast and fibroblast cultures. The results showed that PGE2 and ketorolac stimulated intracellular ALP activities of osteoblasts in dose dependent fashions, while very low ALP activities were detected in either the control or agents treated cultures of fibroblast. These results suggest that PGE2 may be involved in osteoblastic differentiation and the stimulatory effect of ketorolac on osteoblastic ALP activity may not be PGE2 mediated. The responses of osteoblasts to both agents can be as the characteristics of primary osteoblast derived from rat calvaria. PMID- 10375867 TI - Detection of human papilloma virus and Epstein-Barr virus DNA in nasopharyngeal carcinoma by polymerase chain reaction. AB - Nasopharyngeal carcinoma (NPC) is an unique epithelial malignancy which occurs at a high frequency in certain regions of Southeast Asia. Human papilloma virus (HPV) and Epstein-Barr virus (EBV) have both been identified in tissue specimens from NPC. Nevertheless, the association between viral infection and NPC remains unclear. The purpose of this study was to demonstrate that simultaneous infection with EBV and HPV can occur in NPC. Eighty-eight fresh tissue samples which contained sufficient and adequate DNA from patients with histologically confirmed NPC were examined for the existence of HPV and EBV DNA by polymerase chain reaction (PCR). The results showed that HPV and EBV DNA were detected in 51% and in 83% of the specimens, respectively. Coexistence of EBV and HPV in NPC was found in 42% of the samples. The "high risk" types including HPV-16 and HPV-18 accounted for 67% of 45 HPV positive samples. Furthermore, 80% of HPV-16 or HPV 18 positive samples also contained EBV DNA. Our findings suggest that coexistence of EBV and "high risk" HPV-16 or HPV-18 infection may be an important factor in the pathogenesis of NPC. PMID- 10375868 TI - HLA-DMA and HLA-DMB genotyping in patients with rheumatic diseases. AB - To investigate the correlation of HLA-DMA and DMB alleles to some rheumatic diseases, HLA-DMA and DMB genes were detected in 11 patients with juvenile rheumatoid arthritis (JRA), 22 patients with psoriatic arthritis, 26 patients with Behcet's disease, 62 patients with ankylosing spondylitis (AS), and 138 unrelated healthy controls. There was no significant difference in phenotypic frequencies of HLA-DMA and DMB alleles between controls and patients with these rheumatic diseases. HLA-DMA and DMB genes are not related to the susceptibility of JRA, psoriatic arthritis, Behcet's disease, and AS. PMID- 10375869 TI - Attenuation of basilar artery spasm after experimental subarachnoid hemorrhage in rabbit by potassium channel activator cromakalim--a preliminary result. AB - Cerebral vasospasm associated with aneurysmal subarachnoid hemorrhage (SAH) remains a major complication in patients suffering from SAH. Regulation of membrane potential of arterial smooth muscle through activation or inhibition of potassium (K+) channel activity provides an important mechanism to dilate or constrict arteries. The present study examined the effect of a K+ channel activator, cromakalim, on cerebral vasospasm following experimental SAH. By the route of topical application and intra-arterial injection, basilar arteries were exposed transclivally and measured on-line using videomicroscopic camera. Continuous microinjection from right vertebral artery was given after the result of application was observed. Basilar artery spasm induced by SAH was released by topical or intra-arterial administration of cromakalim, and this beneficial effect against cerebral vasospasm was dose-dependent. There was no significant difference between topical and intra-arterial administration of cromakalim. These results indicate that K+ channel activator may play an important role for ameliorating cerebral vasospasm. An important goal of future studies will be to carefully evaluate the possibility and effect of intra-arterial administration of cromakalim to treat angiographic vasospasm. PMID- 10375870 TI - Vaccine storage practices in primary care physicians' offices in Taiwan. AB - To assess vaccine storage practices in Taiwan's clinics, an evaluation was made to determine whether they follow the relevant guidelines. A cross sectional study was carried out in March 1994, in which sixty-three clinics were selected at random from the 744 clinics in the twenty-one counties of Taiwan. Vaccine storage practices were inspected and the temperature of the central core of refrigerators used by these clinics for vaccine storage was measured. Of the 63 clinics, the majority (93.7%) stored articles other than vaccines in their vaccine refrigerators. Thirty clinics (47.6%) placed simple thermometers in their refrigerators; twenty-nine clinics (46.0%) used recording thermometers to produce temperature charts; and 25 clinics (39.7%) equipped their refrigerators with UPS (uninterruptable power system). Forty (63.5%) of 63 vaccine coordinators who were interviewed indicated at least one power failure per year. All refrigerators used for vaccine storage were of the domestic type without "safe zone" gauges. We discovered inappropriately high temperatures (> 8 degrees C) in 22% (14/63) of all refrigerators. Most refrigerator doors (76.3%, 45/59) or freezing rooms (73.7%, 28/38) were not maintained at a temperature required for storing vaccine. Coordinators should be carefully trained and refrigeration and electrical facilities be reassessed to improve vaccine storage in primary care physicians' offices. PMID- 10375871 TI - The influence of rehabilitation therapy on the prognosis for stroke patients--a preliminary study. AB - The purpose of this prospective study was to investigate the influence of rehabilitation therapy on the prognosis for stroke patients. Sociodemographic and clinical factors were collected in a sample of 147 stroke patients (81 men and 66 women) admitted to the inpatient rehabilitation department at our university hospital over 10 days between January 1, 1997 and December 31, 1997. Functional Independence Measure (FIM) scores at discharge and gains during rehabilitation period were used as the prognosis index. Statistical techniques with univariate and multiple regression analyses indicated that significant predictors of discharge FIM scores include age, previous attacks twice or over, medical comorbidities, sensory and orientation impairments, and dementia. In addition, previous stroke attacks twice or over and sensory impairment were significant predictors of FIM gains during rehabilitation period. We concluded that: 1) age is a critical factor to determine the rehabilitation outcome, but may not be an important factor to predict the ability for the improvement through rehabilitation therapy; 2) the delay of rehabilitation therapy may not affect the potential for further improvement; 3) patients with low initial functional level may have poor final outcome, they may still have good rehabilitation potential to improve the functional level; 4) complications of stroke may affect the rehabilitation outcome and should be prevented; and 5) patients with impaired mental status should not routinely be excluded from rehabilitation programs. PMID- 10375872 TI - An indigenous melioidosis: a case report. AB - Melioidosis is a rare but potentially fatal infectious disease in Taiwan, although it has been endemic in Southeast Asia, especially northeast Thailand, and northern Australia. In this article, we report a male diabetes with fulminant pneumonia, and septicemia caused by Burkholderia pseudomallei without traveling abroad before this episode. Productive cough and intermittent chills, high fever for one week, followed by progressively deteriorating dyspnea, shock, disturbed consciousness status were the major presentations. Blood culture grew B. pseudomallei on the fifth admission day. Unfortunately, the patient died on the 9th admission day, despite intensive care and the broad-spectrum antimicrobial regimen used. PMID- 10375873 TI - Rapid progression of parkinsonism associated with an increase of blood manganese. AB - In this paper, we report a 72-year-old man whose parkinsonian pictures accelerated rapidly after an ingestion of unknown herb pills. His serum manganese and aluminum level increased 2-fold and 5-fold over physiological level respectively. A reverse of his parkinsonian deterioration was accompanied with a normalization of these metals. Exclusive heavy metals have been widely mentioned in parkinsonism. While industrial source of these metals has extensively been sought, pharmacology is rarely mentioned in this aspect, especially of herb medicine origin. We suggest that an acceleration of parkinsonian pictures should raise the need to re-evaluate the possibility of heavy metal intoxication in parkinsonism. Besides of industrial contamination, we should be alert for the nonindustrial source in our population. PMID- 10375874 TI - Diabetes insipidus in Langerhans' cell histiocytosis: report of a case. AB - The incidence of diabetes insipidus secondary to Langerhans' cell histiocytosis (LCH) varies among different reports, ranging from 9.5 to 50%, but it has never been reported in literature in Taiwan. Therefore, we presented a case suffering from polyuria, polydipsia, body weight loss for more than one year and seborreic dermatitis-like skin lesions over the scalp and trunk for more than two years. Her body weight and body length were both less than 3 percentile. Fluid restriction and vasopressin test were performed to differentiate nephrogenic from neurogenic diabetes insipidus. Skin biopsy revealed picture of LCH and LCH with complete central diabetes insipidus was diagnosed. Brain MRI and other laboratory examinations were all within normal limits. She received nasal DDAVP treatment and chemotherapy with TPOG-H 94 protocol. After 3 months treatment, her skin lesions disappeared and daily urine amount returned to normal range. PMID- 10375877 TI - The promise of medical informatics. PMID- 10375876 TI - Report from HIMSS. Information tools as enablers. PMID- 10375878 TI - Health informatics and the Hospital Ethics Committee. PMID- 10375879 TI - Taking HL7 to the next level. AB - For HL7 to reach the next level of standards capability, the organization must continue their model-centric development process so systems using the standards can achieve interoperability--sematic as well functional. The HL7 Message Development Framemwork specifies thi process detail and anchors future HL7 standards with a common reference information model. That model will evolve through a process of discussion, amendment, voting to continuously harmonize proposals from committees that have specific spheres of knowledge. PMID- 10375880 TI - Getting to the core of medical informatics training. PMID- 10375881 TI - System analysis and design in five easy steps. PMID- 10375882 TI - The Columbia medical informatics story: from clinical system to major department. PMID- 10375883 TI - Conference report. Medicine meets virtual reality. PMID- 10375884 TI - Assessing the impact of ambulatory computer-based medical record systems. PMID- 10375885 TI - Medical knowledge on demand. Highlights from the third annual DMI conference. PMID- 10375886 TI - Catching the wave. Breakthroughs in wireless technology. PMID- 10375887 TI - SMPTE test pattern evaluation of teleradiology. Using data compression and a standard modem. PMID- 10375888 TI - Clinical practice guidelines on the Internet. A structured, scalable approach. PMID- 10375889 TI - The utilization of a neural network system in the diagnosis of appendicitis. PMID- 10375890 TI - The evolution of computer programming. PMID- 10375891 TI - [The 100th Congress of the Japanese Society of Otolaryngology. Sendai, Japan. May 20-22, 1999. Abstracts]. PMID- 10375892 TI - [Wernicke's encephalopathy]. PMID- 10375894 TI - [Diabetic neuropathy]. PMID- 10375893 TI - [Dementia associated with metabolic diseases]. PMID- 10375895 TI - [Neurologic dysfunctions due to poisoning]. PMID- 10375896 TI - [Mitochondrial abnormalities and nervous dysfunctions]. PMID- 10375897 TI - [AIDS encephalopathy]. PMID- 10375898 TI - [Nervous system dysfunctions due to collagen diseases]. PMID- 10375899 TI - [Nervous system diseases associated with sarcoidosis]. PMID- 10375900 TI - [Neuro-Behcet disease]. PMID- 10375901 TI - [Autoimmune nervous dysfunctions]. PMID- 10375902 TI - [Sleep apnea syndrome]. PMID- 10375903 TI - [Coagulopathy and cerebrovascular disorders]. PMID- 10375904 TI - [hypertensive encephalopathy]. PMID- 10375905 TI - [Heart diseases and cerebrovascular disorders]. PMID- 10375906 TI - [Neurologic diseases associated with malignant tumors]. PMID- 10375907 TI - [Nervous system dysfunctions associated wtih systemic diseases: discussion]. PMID- 10375908 TI - [Case of Cushing's syndrome due to bilateral adrenal adenomas with one presenting as melanotic adenoma]. PMID- 10375909 TI - [Case of infective aortitis initially developing marked edema of the legs and resulting in sudden death due to rupture of the right coronary artery]. PMID- 10375910 TI - [Case of ANCA-related nephritis complicated with central retinal vein occlusion]. PMID- 10375911 TI - [Chronic hepatic dysfunction with fatal outcome due to Vibrio septicemia: report of 3 cases]. PMID- 10375912 TI - [Case of small-cell lung carcinoma associated with Lambert-Eaton myasthenic syndrome]. PMID- 10375913 TI - [Collagen diseases and hemophagocytic syndrome]. PMID- 10375914 TI - [Repair and regeneration of renal tissue]. PMID- 10375915 TI - [Hypertension therapy and molecular biology--with special reference to renin angiotensin system]. PMID- 10375916 TI - [Contemporary model for treatment of erythroplakia]. AB - In literature worldwide is still commonly used the term erosion to describe red areas within cervix around the external orifice. In such cases with negative cytology result, for cervical cancer prevention, the electrocoagulation or electro-conisation or other destructive operations are routinely used. Without colposcopy verification such management is inappropriate. The physician treats but does not know what he treats. It may be both common ectopy or regeneration zone so physiologic cervical states but it may be also CIN or even early cervical cancer, however cytologically negative. The first group of lesions is effectively diagnosed with colposcopy without additional diagnostic procedures and the CIN lesions are diagnosed in high percentage of accuracy. Not all of these lesions should be treated. In the group of colposcopically and cytologically unsuspected lesions just very extensive lesions with active mucous glands should be treated. Such lesions cause recurrent cervical inflammation. All other erythroplakia type lesions demand no treatment. The presence of ectopy around the external cervical orifice is just profitable for diagnosis of epithelial changes and cervical physiologic processes observation. All cases of abnormal colposcopy or cytology results, suspected of CIN should be treated as prevention of cervical cancer. In lower CIN grades electroresection (LEEP) is recommended, while in higher grades the cervical conisation is the appropriate mode of treatment. PMID- 10375917 TI - [Colposcopy screening for prevention of uterine neck neoplasms]. AB - Cervical cancer, throughout the world, is the second most common neoplasm among women. In Europe approximately 22,000 new cases of cervical cancer are diagnosed each year. This study assessed colposcopy as a screening method in the detection of pre- and early neoplastic lesions. This retrospective study analysed 3264 women hospitalized or treated on an outpatient basis at the Gynaecology and Oncology Clinic of CMUJ from 1995 to 1997. Among 81.2% patients the unsuspected colposcopic pictures were found which were histologically verified. In 5.9% of all examined patients the colposcopic pictures suggested lesions of CIN and early invasive cancer. In these cases the colposcopy presented false positive pictures in 2.6%. From this analysis the colposcopy is a method of good concordance with histologic examination result. This allows to eliminate unnecessary punch biopsies and even diagnostic conisation in significant percentage. PMID- 10375918 TI - [The role of colposcopy in evaluation of the cervix for unexpectedly positive pap smears]. AB - The cervical cancer is still one of the most common malignancies of female low genital tract (LGT). Pap smears is a widely used screening procedure. Colposcopy with punch biopsy verifies positive Pap smears. Clinical material were 133 women aged 34-75 yrs diagnosed and operated in the Chair and Department of Gynecology and Oncology CM Jagellonian University. In all cases cytology and colposcopy were performed. Their results were compared to each other and to histology results. The concordance of cytologic and histology results reached 92.5% while of colposcopic and histology--98%. Colposcopy is an useful method of Pap smears verification because it limits to minimum false negative results percentage. PMID- 10375919 TI - [Incidence of precancerous states an carcinomas of the vulva in young women]. AB - Frequency of precancerous and malignant neoplasms in young (under 45 y.) women in last 20 years has been analysed in the work. There have been analysed VIN and invasive carcinoma progression, localization, coexistence of HPV infection together with intraepithelial neoplasia within uterine cervix and vaginal mucosa. 50% increase of VIN and vulvar cancer, especially in young women during last 10 years has been found. Earlier clinical stadium of vulvar carcinoma is usually found in this group of women compared with older ones. There has been found high percent of multi-focal localization of the vulvar changes in younger patients, especially as far as VIN is concerned. It is often accompanied by HPV infection (60.5%), and rarely by dystrophic changes (20.6%). 11.8% coincidence of cervical intraepithelial neoplasia and vaginal intraepithelial neoplasia, mostly accompanied by HPV, with VIN and vulvar cancer, has also been found. PMID- 10375920 TI - [The management of cervical cancer cases in pregnancy and puerperium]. AB - Cervical cancer in women during pregnancy and puerperium is a serious diagnostic and therapeutic problem. Twelve multiparas with confirmed cervical cancer during pregnancy, delivery and puerperium were examined. The mean age of the group was 35. In two of them cervical cancer was diagnosed in the second trimester, in 5 in the third trimester and in 5 in puerperium. Clinical stage according to FIGO was as follow: Ib--9 patients, IIa--2 patients, III--1 patient. In two patients operated in the second trimester--extended hysterectomy was performed. In four women cesarean section with extended hysterectomy and lymphadenectomy was performed. Only one patient in third trimester had cesarean section and in the same time unradical hysterectomy because of bleeding. In two patients in puerperium extended hysterectomy was performed (Meigs operation). Three patients underwent only radiotherapy. All patients who were operated on underwent subsequent radiotherapy. CONCLUSIONS: Cervical cancer during pregnancy and puerperium is diagnosed very late, usually in advanced stage. It is connected with lack of clinical and cytological examination of women before pregnancy. Principles of treatment of cervical cancer in pregnancy and puerperium do not differ from those applicable in other patients. PMID- 10375921 TI - [The role of phase contrast microscopy in the evaluation of cervix neck and vaginal biocenosis]. AB - The aim of our study was to evaluate and compare the possibility of germ identification in cervico-vaginal infections of different origin by Ph cytology and classic fixated and stained (H-E) smears. We identified Lactobacillus vag., Gardnerella vag., short bacilli (Escherichia coli, Proteus, Klebsiella), mixed Coccus flora (diplococci, small and large cocci), Trichomonas vag., and Candidia alb. in cervico-vaginal smears. In endocervical, mature metaplastic or parabasal cells the elementary and cocoid bodies of Chlamydia trachomatis were possible to observed. The cytopathic effects caused by viral infections (HPV, HSV, CMV) were detected. The classic (koilocytosis, dyskeratosis) and discrete (binucleation, cracked cells, ghost cells, tadpole cells and large cytoplasmatic hyaline granulations) morphological changes must be confirmed by colposcopy and Digene Hybryde Capture analysis. The results of identification of vaginal biocenosis in Ph microscope and H-E smears were similar. The main benefit of Ph cytology is accuracy, speed and reliability of comprehensive information. PMID- 10375922 TI - [Integrated colposcopy and phase contrast microscopy for diagnosis of squamous intraepithelial lesions and invasive cervical neoplasms]. AB - Colposcopy or cytology examinations alone allows to obtain diagnostic concordance with histopathological examination between 74-89% and 68-86%, respectively in SIL and cervical cancer detection. Both methods used simultaneously allow increase concordance with histopathology up to 95-100%. Cytological examination may be used simultaneously with colposcopy when Ph microscopy is used. The aim was to estimate benefits of conjunction of colposcopy and Ph cytology in SIL and cervical cancer detection in daily gynecological practice. Integrated colposcopy and cytology examinations were performed in 200 patients. Cervico-vaginal smears were obtained from the patient before Acetic-Acid Test and Lugol-Solution-Test were performed. Wet smears (Ph) were made. Results of integrated colposcopy and cytology were verified by histopathological examination. Diagnostic specificity, sensitivity, and Youden's value were estimated for Ph cytology, H-E cytology, and colposcopy alone and for integrated colposcopy and Ph cytology. Simultaneous colposcopy and Ph cytology reveal specificity 97.1% sensitivity 94.6% and Youden's value 91.8% thus provide fast, reliable and comprehensive information for accurate diagnosis in daily gynecological practice. PMID- 10375923 TI - [Colposcopic verification of cytologic smears containing atypical squamous cells of undetermined significance]. AB - The aim of the study was the estimation of significance of smears containing Atypical Squamous Cells of Undetermined Significance (ASCUS) and presentation the own management of that cases. The study group consisted of 73 non-pregnant women (aged from 21 to 54 years) with ASCUS. The control group included 113 non pregnant patients (aged between 20 and 57 years) with unsuspected cytology findings. Chi square test and Fisher test were used to compare the colposcopy findings in the two groups. Colposcopic pictures indicating a presence of dysplasia were significantly more often in study group. 14 cases (19.18%) of mild dysplasia and 8 cases (10.96%) of moderate and severe dysplasia were found in the group of patients with ASCUS. During the 18 month follow-up of the remaining patients in that group persistence of ASCUS was found in 2 cases (3.92%), progression to mild and moderate dysplasia was observed in 6 women (11.76%). Regression to normal smear occurred in 43 women (84.31%). Progression to dysplasia referred to women with suspected colposcopy findings. Until further research resolve all the questions regarding ASCUS smears or new more precise diagnostic methods are invented, colposcopy remains the method of choice in verification of ASCUS smears. Colposcopy together with cytology is the optimal mode of observation of such patients. PMID- 10375924 TI - [The value of micro-colpo-hysterectomy in the diagnosis and therapeutic evaluation of squamous intraepithelial lesions]. AB - Simultaneous application of cytology, colposcopy and colposcopically directed punch biopsy is acknowledged diagnostic procedure in squamous intraepithelial lesions (SIL) and cervical cancer, but not in every case it allows for precise estimation and localisation of the lesion. The aim of the study was evaluation of the value of microcolpohysteroscopy (MCH) in diagnostics and therapeutical qualification of SIL. The study group comprised 86 women cytologically suspected for SIL. Sensitivity, specificity, positive and negative predictive values of MCH in detection of SIL were calculated. The estimation of the value of MCH in therapeutical qualification was based on therapy effects of SIL performed according microcolpohysteroscopy evaluation of the cervix. In cases of low-grade SIL the sensitivity of MCH reached 0.73, the specificity 0.5, positive predictive value was 0.85 and negative predictive value was 0.33. For high-grade SIL and cervical cancer the values were 0.98, 0.33, 0.95 and 0.5 respectively. Totally, for all lesions of SIL type and cervical cancer sensitivity of MCH reached 0.94, specificity 0.75, positive predictive value was 0.99, and negative predictive value was 0.38. The high value of microcolpohysteroscopy in evaluation of the cervix made possible to optimize the treatment mode and adequately employ of minimal invasive therapeutic methods. PMID- 10375925 TI - [Results of cervix neoplasm prophylaxis in women examined during the health promotion program in the Central Railway Hospital]. AB - In the program of cervical cancer prophylaxis in CRH in 1997 simultaneous cytology and colposcopy evaluation of the uterine cervix were performed. Among 1479 women examined there were 6 cases (0.4%) of CIN 3, 34 cases (2.79%) of CIN 1 and CIN 2 and no case of invasive cervical cancer. The results of these studies point at high effectiveness of simultaneous use of colposcopy and cytology as complementary methods in prophylaxis of cervical cancer. PMID- 10375926 TI - [Correlation between oncologically suspicious cytologic smears and colposcopic images of the uterine cervix and results of histopathologic examination taken from material provided by the Gynecology Department of the Pruszkow Municipal Hospital from 1995 to 1997]. AB - The objective of the paper was an evaluation of relationship between oncologically suspicious cytologic smear and colposcopic images, and the results of histopathological examination of samples collected from the vaginal portion of uterine cervix and from cervical canal scrapings. The material consisted of 60 patients with cytologic group 3 and 13 patients with group 4 and 5 according to Papanicolaou, who had been selected during a two-year preventive examination period. Cytologic group 3 patients (group 1) were subjected to cytological assessment and colposcopic examination again in order to determine further diagnostic indications. Cytologic group 4 and 5 women (group 2) were collected samples from the vaginal portion of uterine cervix under colposcopic supervision, without waiting for the result of cytologic re-assessment. All these patients were also collected samples from the vaginal portion of uterine cervix. In patients of cytologic group 3 the cytologic and colposcopic examination results were found to be compatible with histopathologic examination results in 53.3% cases. In patients of cytologic group 4 or 5 this factor was established at 92.3%. PMID- 10375927 TI - [Appearance and evolution of cytologic changes of squamous epithelial cervical cells during the course of human papillomavirus infection. I. Progression of cervical dysplasia]. AB - There was planed a cycle of works devoted to the title problem. The aim of the first of them was evaluation in our material influence of HPV infection on dysplasia progression of cervical squamous epithelium in correlation to the detected HPV type. PMID- 10375928 TI - [Histopathologic status of endocervical canal epithelium in CIN changes localized to the ectocervix]. AB - The histopathologic assessment of endocervical canal epithelium in the presence of ectocervical CIN is a major determinant of therapy. The presence or absence of neoplastic changes within the endocervical canal is one of the main modifying factors of the therapeutic approach. In the group of 97 women examined colposcopically, ectocervical punch biopsies of the cervix and endocervical curettage samples were obtained. Patients were divided into two groups based on colposcopic ectocervical appearance. The first group of 44 patients had a mildly suspicious colposcopic ectocervical appearance. The second group of 53 cases had a highly suspicious colposcopic ectocervical appearance. A comparison of the histopathological results of ectocervical punch biopsies with those of the endocervical curettage samples presented the following: in 44 women with a mildly suspicious colposcopic ectocervical appearance, 5 (11%) had CIN type endocervical lesions. Of 57 women with a highly suspicious colposcopic ectocervical appearance 13 (24%) had endocervical CIN type lesions. Curettage of the endocervical canal is an equally important part of the assessment of CIN type changes on pair with ectocervical punch biopsy. This is particularly true in cases of a highly suspicious colposcopic ectocervical appearance. PMID- 10375929 TI - The use of carbon-dioxide laser surgery in the treatment of intraepithelial neoplasia of the uterine cervix. AB - From October 1989 to June 1997, 1841 patients (pts) suffering from different diseases of the lower genital tract have been treated with CO2 laser surgery in our Institution: among them, 782 were affected by cervical intraepithelial neoplasia (CIN). All pts underwent CO2 laser procedure for CIN after adequate colposcopic evaluation of the entire lower genital tract, colposcopic guided biopsy of the lesion, adequate pre-surgical work-up for possible infectious and coagulation associated disease. In 736 (94.1%) pts, the procedure was performed on an ambulatory basis while 46 pts (5.9%) were admitted for 1 or 2 days. A CO2 laser Sharplan 55 associated to a Zeiss operative colposcope was employed. The preoperative diagnosis of the 782 pts treated for CIN was 297 CIN1, 255 CIN2, 171 CIN3 and 59 CIS. Mean age was 33.6 years without statistical difference among the grade of disease: 605 pts underwent laser vaporization according to specific selection criteria. The depth of cervical destruction was less than 6 mm in 26 cases, between 6 and 10 mm in 549, between 11 and 15 mm in 157, more than 15 mm in 50 pts. 742 procedures were performed under local anesthesia and pain was absent in 667 pts. (89.9%), mild in 51 (6.8%), moderate in 19 (2.5%) and severe in 5 (0.7%). Intraoperative bleeding was severe in 30 pts. (3.8%), moderate in 77 (9.8%), mild in 204 (26.1%) and absent in 471 (60.2%). The conization procedure was shown to have a higher risk of bleeding but no direct relation was observed with the depth of cervical destruction. Late complications were scarce: 1.3% of late hemorrhages, 1.4% of stenosis of cervical external orifice and cervical endometriosis in 0.3%. In 76 pts (42%) of the 177 conizations the final pathology report was in accordance with the previous biopsy, in 56 (30.9%) a lower grade of CIN was observed, in 53 (29.3%) a worse grade of the lesion was retrieved. Among these latter pts. 10 showed a microinvasive and 2 an invasive cancer: both the invasive but only 3 of the 10 microinvasive cancer pts underwent a surgical procedure (2 radical and 3 extrafascial hysterectomies, respectively). After a mean follow up of 37 months our incidence of recurrence is 2.3% (18 pts): 5 CIN1, 7 CIN2, 3 CIN3, 2 CIS and 1 microinvasive disease. In 78% of the instances the recurrence has occurred within the first year of follow up. All 18 recurrences were successfully treated with further vaporization in 8 cases, conization in 9 and hysterectomy in 1. 93 term pregnancies occurred in 83 pts after CO2 laser treatment of CIN. No cervical incompetence occurred (no cervical cerclage employed) while the incidence of spontaneous abortion was not statistically significant. 82 pregnancies were delivered vaginally without significant variation of labor phase duration. The incidence of caesarian section (11.8%) was lower than the mean incidence in our Institution. These data confirm the successful complete restitutio ad integrum of the cervix after an adequate CO2 laser surgical procedure without any further risk of cervical incompetence, premature delivery or premature rupture of membranes. The use of CO2 laser surgery is recommended as modality treatment of choice in the management of cervical intraepithelial neoplasia. PMID- 10375930 TI - [The role of preoperative brachytherapy in the treatment of early invasive cervical cancer]. AB - The role of preoperative brachytherapy in the treatment of cervical cancer are discussed. Authors described a group of 45 patients with preliminary diagnosis of Io and IIo A sec FIGO tumour. Qualification system to combined treatment and diagnostic procedure are described. Authors discussed methods of LDR therapy. Tubes of 226Ra in Paris technique and afterloading 137Cs Selectron LDR/MDR (of Nucletron BV) were used. Postoperative histopathologic results are compared to preoperative diagnoses established on the basis of clinical examination, US and MRI. Preliminary assessment of tolerance and efficacy of these methods of treatment is made. PMID- 10375931 TI - [LEEP in gynecologic practice]. AB - The experience in performing LEEP electroconisation, the gaining popularity method of treatment in preneoplastic lesions of the uterine cervix was presented. The simplicity, safety and low number of complications of this out patient procedure was stressed. The attention was paid on diagnostic-therapeutic aspect of the procedure, the adequacy of histological specimen assessment, as well as on the excellent patient's toleration. The weak impact on anatomy and physiology of the uterine cervix was also pointed. The introduction of LEEP conisation to gynecological clinic bears in author's opinion simplification and cost effectiveness of diagnosis and treatment in selected cervical lesions. PMID- 10375932 TI - [The effectiveness of conservative treatment of cervical lesions using the LLETZ and CO2 laser]. AB - Development made on the carcinogenesis process of the cervical lesions and increased detection of the early precancerous lesions enable discontinuance of radical treatments for non-radical techniques which it is of vital importance to young women of the child-bearing capacity. The aim of this study was to determine the efficacy of the non-radical treatment of the cervical lesions using LLETZ procedure (Large Loop Excision of Transition Zone) and laser CO2 vaporisation. 2046 women aged 18-46 who were diagnosed for cervical lesions were treated in the Institute of Obstetrics and Gynaecology Medical School of Bialystok in the years 1994-97. 216 of which were histologically confirmed for CIN I-III diagnosis. The choice between LLETZ or laser CO2 was made based on a pre-treatment examination (cytology, colposcopy, microbiology test and punch biopsy). The final results were evaluated from 6 months to 4 years after the treatment. The effectiveness of CO2 laser was 94.6% and was similar to LLETZ--96.4%. In spite of almost complete agreement in both procedures, the LLETZ seems to be more preferred because of the possibility of histological post-treatment verification. PMID- 10375933 TI - [Operative treatment of early cancerous stages and a woman's quality of life]. AB - The operative treatment of early cervical cancerous stages changes the womans quality of life. In the literature there exist works on influence of radical treatment of invasive cervical cancer on the psychical and psychological state of the operated woman. There are few works on the influence of early cancerous stages and social activity of woman. The aim of this work is the assessment of influence of precancerous and cancerous cervical disease of psychical state, partnership, family life and professional and social activity of woman. The material were 153 women aged 20-47 years diagnosed and treated in the Department of Gynaecology and Oncology CM UJ Krakow in the period 1989-1994 for CIN 3 and the cervical cancer in IA stage. During the hospitalisation the women were tested psychologically and were performed special examination with originally constructed formula in 3, 6 and 12 month after the treatment. There has been stated that the precancerous and early cancerous disease diagnosis causes the disease awareness in these women who stated themselves as healthy, causes the fear of the life and saving the reproductive organ and protection the reproduction ability, their further family, professional activities i.e. woman's quality of life. PMID- 10375934 TI - [Neurotrophic basis of atrophic changes in the vagina and vulva]. AB - The aim of the study is the analysis of morphology and biochemistry of neurovascular peculiarities of biologically active cells of connective tissue and epithelium. Surgically treated women were examined in 3 age groups--active sexual cycle, menopause and senium. Histological-histochemical, neurohistological and neurohistochemical methods and techniques were applied. Morphological pictures of connective tissue proper and epithelium reflect the physiological retroplasia process, associated with age-dependent structural and cellular transformations. This is manifested in typical peculiarities observed in the peripheral innervation and in vascular rete, quantitative and localization changes in mastocytes, chromaffin cells, macrophages, lymphocytes and leucocytes. Quantitative and qualitative rebuilding takes place in fibrous and biochemical structures of basic intracellular substance with its hyalinization. This process of simple tissue atrophy includes pathological activation. In innervation and vascularization, active restructuring coexists with the pictures of quantitative and localization changes in biologically active cells in the series of hormone immune reactions. This is expressed by dysplasia of innervation structures with angiogenesis, while in cells, apart from apoptosis or necrosis, by the appearance of stratified epithelium (anaplasia) in the reproductive layers. PMID- 10375935 TI - [Iscador QuS and human recombinant interferon alpha (Intron A) in cervical intraepithelial neoplasia (CIN)]. AB - For several years there has been the association between the persistent HPV infection (especially with high oncogenic potency i.e. 16, 18) and the cervical intraepithelial neoplasia. The pathomechanism is probably considered with spread of the early virus gene E1, E2 and the suppressor protein p53 complexes. Further on these complexes cause the neoplastic cell transformation. There has also been described the role of impaired immune response in these cases. The abnormalities cover malformation of antigen presenting system APC, decrease of MHC-I and MHC-II heavy chains rate, decrease of the Langer-hans cells and decrease of count and cytotoxic activities of lymphocytes B and NK cells. The invasive and destructive techniques of HPV associated CIN treatment do not respect its pathogenesis. Therefore the new non surgical methods of treatment would play a major role in treatment and prevention of women especially in their reproductive period. The aim of this work was the evaluation of the Iscador QuS and Intron A role in the management of HPV associated CIN. The 60 patients with CIN and HPV have been diagnosed and treated in our clinic for 12 months. Early results present increase of regression and significant decrease of progression rates in both groups of examined women, comparing to the control group. The stationery state rates in this groups of women were similar to the control group. PMID- 10375936 TI - [Tactical diagnostic-therapeutic procedures in early stages of vulvar cancer]. AB - The objective of this report is to present the diagnostic and therapeutic tactics in early vulvar carcinoma i.e. carcinoma in situ and carcinoma in I stage (IA and IB) which aim is to limit the radicalisation of the surgery. The clinical material is 57 women aged 27-63 years, colposcopy, cytology and histology diagnosed. The histology examination was made under colposcopic control, and the initial staging was settled i.e. ca in situ in 11 cases, carcinoma IA in 7 cases and carcinoma IB in 39 cases. In qualifying criteria for surgery, unifocal or multifocal location in carcinoma in situ was analysed, and presence of dystrophic lesions in ca in situ and ca IA was taken into consideration. In ca IB the additionally diameter of the lesion (1 cm and 1-2 cm), location (clitoris, perineum, or pudendal lip) and in carcinoma located within the pudendal lip, presence of metastases in lymphatic nodes of inguino-femoral region was taken into consideration. Local excision of lesion was therapeutical procedure in majority of cases in carcinoma in situ and carcinoma IA stage. In multifocal localisation of ca in situ or in coexistence of dystrophic lesions in ca in situ and ca IA vulvectomy was performed. But in carcinoma IB located within the pudendal lips, where lack of metastases to lymphatic nodes in intraoperative histopathology was confirmed, local excision of neoplastic lesion was performed. Presence of lesion within the clitoris, perineum or presence of metastases in lymphatic nodes led to more radical procedure including radical vulvectomy with bilateral lymphadenectomy of inguino-femoral region. PMID- 10375937 TI - [A comparison of clinical and surgical-pathologic staging of advanced vulvar cancer]. AB - A surgico-pathological staging system introduced in 1988 by FIGO replaced previously used clinical classification of vulvar cancer extent. The aim of our study was to compare retrospectively the prognostic value of both of these staging systems (with the modification of surgical staging introduced in 1994) in the group of 123 vulvar cancer patients undergoing radical surgery including lymph node dissection in our institution. The comparison of both staging systems is presented in the table. Ninety patients (73%) have been assigned to the same stage in both systems, in 27 patients (22%) the use of surgical system resulted in upstaging, as compared to the clinical classification and 6 patients (5%) have been down-staged by the surgical system. In the analysis of prognostic value of both classifications the surgical staging system proved to have better discriminating power for prediction of survival in vulvar cancer patients. PMID- 10375938 TI - [Analysis of intra- and postoperative complications and postoperative course in patients surgically treated for vulvar cancer]. AB - Surgical complications were evaluated in 162 vulvar cancer patients. Assessed were: the frequency and type of postoperative complications, the incidence of perioperative blood transfusions, the course of postoperative wound healing, the length of postoperative hospital stay and reasons for perioperative mortality. An analysis of factors influence the risk of complications was conducted. PMID- 10375939 TI - [Wound healing after vulvar surgery]. AB - Vulvular wounds healing is an important problem in vulvar surgery. Prolonged healing worsens patients quality of live, and is related to prolonged hospitalization and use of antibiotics. The aim of this report is to analyse factors, which may influence vulvular wounds healing. It was found that extended vulvar surgery and advanced age of patients are the negative factors in vulvar wound healing. Antibioticotherapy, drainage and diseases like diabetes mellitus, hypertension and obesity don't affect the vulvular wounds healing. PMID- 10375940 TI - [Giant condylomas acuminata of the uterine cervix in a pregnant woman]. PMID- 10375941 TI - [Chordomas. Analysis of 24 cases]. AB - Chordoma is a relatively rare neoplasm originating from the remnants of the embryonic notochord. We analysed 24 cases of chordoma. The patients were divided into groups depending on the localisation. We analysed the results of different methods of therapy: surgical treatment, radiation therapy and combined surgical and radiological treatment. The results were compared with the data from the references. PMID- 10375942 TI - [Clinical observations concerning piracetam treatment of patients after craniocerebral injury]. AB - Piracetam (Nootropil) is a cytoprotective to brain tissue and improving cerebral blood flow medicine. In the Department of Neurotraumatology we investigated results of piracetam treatment in a group of 100 succeeding patients admitted between 1995-96 due to craniocerebral injury. High doses (24-30 g per day) of this medicine have a positive effect on final result of treatment, when treatment is initiated immediately after the injury and described conditions are abided. We also showed usefulness of piracetam treatment in posthospital management. PMID- 10375943 TI - [Evaluation of general medical awareness and extent of preoperative explanation of surgical patients]. AB - In order to help sick persons with making an aware decision to undergo a surgical treatment it is necessary to instruct them in every possible aspect of the proposed therapy. Aiming at adjusting the range and form of information to individual case of each patient, we have tried to estimate his/her general medical awareness and the extent of the acquirement of information received. On the ground of analysis of the results, it was found that 20-50% of patients had no basic knowledge of their illness at the moment of their admission to the hospital. Although 67% of the patients were satisfied with the questionnaire and preoperative explanation, every other patient had no understanding of the essence of the medical action planned. At the same time, about 50% of the patients declared a possibility of withdrawing their consent to surgical treatment, if given more comprehensive information about possible complications. PMID- 10375944 TI - [Diagnosis and treatment approach in anaplastic thyroid carcinoma]. AB - The paper presents diagnosis and treatment of anaplastic thyroid cancer based on the own experience and literature review. There were presented up to date diagnosis and treatment which included radical thyroidectomy and radio-, chemio-, and hormonotherapy. The results of radical thyroidectomy and teleradiotherapy in anaplastic thyroid carcinoma are not satisfactory. PMID- 10375945 TI - [Treatment approach for substernal and intrathoracic goiter. Personal experience]. AB - The retrospective analysis of 489 cases of substernal and intrathoracic goiters among 4122 patients undergoing surgical treatment between 1984 and 1996 due to various thyroid gland diseases including clinical data, surgical technics and early postoperative complications was performed. The surgical procedures of substernal and intrathoracic goiter amounted to 11.9% of all thyroid gland surgery. In 468 (95.5%) patients goiter was situated substernally, in 22 (4.5%) intrathoracicaly. The mean age and time of goiter growth in that location exceeded over 10 years the location of goiter within the neck. In preoperative examination the X-ray of chest and trachea were essential. Routine ultrasonography and thyroid gland scyntigraphy were scarcely helpful as the retrosternal and mediastinal region were often omitted. The jugular access was dominant (98.6%), sternotomy was performed in 1.4% of cases due to big disproportion between size of the goiter and size of the upper inlet into the chest. The surgical complications, similarly as in goiter within the neck (no cases of pneumothorax were observed), included the single-side paresis of recurrent laryngeal nerve in 3.7% of patients, in 0.2% hypoparathyroidism, in 1% bleeding requiring reoperation and in 0.2% esophageal fistula (self-healed). The surgical treatment of retrosternal and intramediastineal goiter was safe and a total number of complications was comparable to that one in a group of patients under-going surgery due to goiter within the neck. Most of surgical procedures was possible to perform using the jugular access. In a small number of cases because of difficulties related to the anatomical conditions the access was reached through the oblong sternotomy. PMID- 10375946 TI - [Acute respiratory failure in the course of thyroid disease]. AB - During the past 10 years, we have treated 11 patients who were admitted with acute respiratory failure due to goiter. Multinodular goiter was examined in four patients, one patient suffered from Graves disease and six patients had malignant thyroid lesions. Four patients required emergency intubation and in two cases we performed tracheostomy because of significant narrowing of trachea lumen. Diagnosis was made in most cases based on chest x-ray films, laryngoscopic examination, CT scans of the neck and goiters fine needle aspiration biopsy. All patients underwent mostly subtotal thyroidectomy with the standard cervicotomy approach. Five patients required tracheostomy procedures after surgery, three because of local advanced malignancy and two because of postoperative respiratory failure. In two cases pneumonia developed as a postoperative complication. One patient with advanced malignant goiter died. The goiters with progress of trachea compression symptoms should be operated in elective surgery to avoid sudden life threatening complications like severe dyspnea. When the hoarseness is present the possibility of malignant goiter is particularly high. In our opinion patients with suspected respiratory failure should be referred to the centres with special interest in thyroid surgery. PMID- 10375947 TI - [Analysis of voice disorders in teachers treated in the Phoniatric Laboratory of the Otolaryngology Department AM in Wroclaw]. AB - Analysis of voice disorders in 898 teachers was performed. In all cases otolaryngologic, videostroboscopic and phoniatric examinations were completed. There was prevalence of women at the age ranged from 46 to 56 years. Majority of morphologic changes included Reinkes oedema (120 cases), chronic simple laryngitis (109 cases), vocal nodules (95 cases), chronic hypertrophic laryngitis (70 cases) and laryngeal polyps (35 cases). In 175 cases vocal insufficiency of glottis and in 57 cases functional disorders were diagnosed. In 131 cases no morphologic and functional changes were found. In the group of teachers the incidence of vocal insufficiency was higher than in control group. The analysis of the examined teachers revealed that professional disease was confirmed in 33% of cases. Majority of them included vocal nodules and chronic hypertrophic laryngitis. PMID- 10375948 TI - [Extracellular components of bronchoalveolar lavage fluid (BALF) as a marker of the activity of interstitial pulmonary diseases. III: Phospholipids to protein concentration ratio]. AB - In interstitial pulmonary diseases investigations are conducted to find markers of the activity of the interstitial processes so that noninvasive monitoring of the disease might be possible. In 188 patients divided into 9 groups: 42 with active sarcoidosis, 24 with inactive sarcoidosis, 16 with active sarcoidosis treated with steroids and 22 with inactive sarcoidosis after corticotherapy, 17 with avian fanciers' lung exposed to the antigen, 16 with avian fanciers' lung after a year interval in exposure to the antigen, 20 with advanced and 13 with moderate idiopathic pulmonary fibrosis, and 18 healthy persons the BAL was performed. In the BALF concentrations of protein and phospholipids were assayed by colorimetric method. The results indicate usefulness of the studied biochemical parameters in BALF in evaluation of the activity of interstitial pulmonary diseases. Significant differences were found between the results in the active group of patients compared to the control group and to the inactive forms of interstitial pulmonary diseases. Particularly valuable is phospholipids to protein concentration ratio in BALF. PMID- 10375949 TI - [Movement efficiency of the cervical spine in pain syndrome associated with instability of the cervical segment of the spine]. AB - The goal of this work is to attempt to evaluate the motor efficiency of the cervical spine by taking into account the range of active movements and the strength of muscle groups in pain syndrome associated with instability of the cervical segment of the spine (CSPS), to point out possible dependencies between the tested parameters of motor efficiency, and to indicate the suitability of motor efficiency examination in assessing the effectiveness of rehabilitation. The results from tests performed on 71 patients indicate that persons suffering from CSPS, in comparison to healthy subjects, ae characterized by very low motor efficiency of the cervical spine. The greatest limitation on the range of active movements occurred in vertical inclination and rotation, while the greatest deficit of strength appeared in the muscle groups of lateral flexors. Moreover, these parameters of motor efficiency showed a high degree of dependency in the examinations. The results from the preliminary examination and the high dynamics of change in the control examinations suggest that motor efficiency examinations are useful in assessing the effectiveness of rehabilitation procedures in patients with CSPS. PMID- 10375950 TI - [Examination of the quality of life in post stroke patients]. AB - The clinical assessment of patients after stroke is usually focused on the detection and determination of neurological deficits. However, the full picture of the patient after stroke should take into account also cognitive deficits, emotional and intellectual disturbances as well as limitations in performing one's social functions. Thus, assessing quality of life after stroke is an alternative to traditional methods of evaluation of the patient's condition. The study presents the attempts to define quality of life determined by the health status and to review questionnaires devised for the examination of post-stroke patients. Despite a considerable variety of methods used, reliable evaluation of all the areas of the patients life after stroke seems impossible using only one research tool. There is also an urgent need to develop an uniform system of evaluation, appropriate for Polish cultural standards. PMID- 10375951 TI - [Post-steroidal osteoporosis]. AB - Pathogenesis of corticosteroid-induced osteoporosis and an influence of several forms of these drugs on bone mineral density were presented. Principles of prevention and treatment of this type of osteoporosis were described. Among the drugs studied, calcitonin, bisphosphonates, calcium, vitamin D and hormonal replacement therapy in women appear to be most consistently effective in increasing bone density. It is also important to modify risk factors of osteoporosis (increased physical activity, diet rich in calcium, discontinuation of smoking, avoidance of excess alcohol intake). PMID- 10375953 TI - [Renal renin-angiotensin system in homeostatic water-electrolyte regulation]. AB - There are many local renin-angiotensin systems in different organs of the body. The renal system is relatively well-known. Localization of particular components of that system, its intrarenal action, influence on arterial blood pressure regulation and sodium homeostasis have been described. PMID- 10375952 TI - [Cotinine--the biomarker of exposure to tobacco smoke]. AB - Long-term exposure to tobacco smoke is conducive to increased morbidity rate of respiratory, alimentary and circulatory diseases. The biochemical and immunochemical studies permitted to assess the basic pathways of nicotine change in mammal organisms. The measurement of cotinine in the system fluids is a specific marker of the intensiveness of smoking, of the exposure connected with active and passive smoking and it can also be of use in nicotine therapy monitoring during the cessation of smoking. PMID- 10375954 TI - [Nonsteroidal anti-inflammatory drugs--nephrotoxic mechanism of action]. AB - Non-steroid anti-inflammatory drugs are easily available and commonly used. Mechanism of their action is based on inhibiting prostaglandin synthesis. Prostaglandins are arachidonic acid derivatives that are responsible, among others, for regulation of renal blood flow. In some kidney disorders as well as in hemodynamic disturbances, their increased release aims at balancing substances causing kidney ischemia. Blocking prostaglandin synthesis in such conditions may result in development of nephrotoxic effect, manifesting in water-electrolyte imbalance, acute tubulo-interstitial nephropathy, nephrotic syndrome, acute and chronic renal papillary necrosis as well as acute or chronic renal failure. Analgesic nephropathy with papillary necrosis is a particular form of the nephrotoxic effect of non-steroid anti-inflammatory drugs. Development of this complication has been described in patients abusing phenacetin or other analgesic drugs and especially their combination. PMID- 10375955 TI - [Anaphylactic shock leading to death in a young woman after oral administration of metamizole (Pyralginum-Polfa)--case report]. AB - The purpose of the report was to describe a case of anaphylactic shock in a 22 year-old woman after administration of one tablet of metamizole (pyralginum Polfa). The patient died, notwithstanding intensive treatment applied rapidly. The patient had been treated for bronchial asthma for a few years. There had been no side effects after repeated administration of metamizole before. PMID- 10375956 TI - [Recurrent ventricular fibrillation in a patient with Prinzmetal angina pectoris- case report]. AB - The description of the case of 35 years old patient with paroxysmal ventricular fibrillation in a course of Prinzmetal angina pectoris treated unsuccessfully with antiarrhythmic drugs who required implantation of cardioverter/defibrillator. The via-venous cardioverter/defibrillator was implanted (ICD). During 6 months observation ventricular tachycardia and ventricular fibrillation occurred four times and was effectively interrupted by the cardioverter/defibrillator. PMID- 10375957 TI - [Past and present at the Polish Medical School in Edinburgh]. AB - The article describes the history of the Polish Medical School at the University of Edinburgh, the world-wide unprecedented enterprise. Profiles of some distinguished Polish people or Scots of Polish descents associated with the Polish Medical School are presented. PMID- 10375958 TI - [The last unpublished paper of Ludwik Bierdowski]. AB - In the last year of his life professor Ludwik Bierkowski (1801-1860) introduce a new method of treatment for open fractures, which was not published because of his untimely death. This method was described by his son Wlodzimierz Bierkowski (1832-1888) but it is still in manuscript. However the method was used by his students. PMID- 10375959 TI - [Magnesium (Mg) status in patients with cardiovascular diseases]. AB - Mg is the fourth most abundant total cation in the human body and the second most abundant intracellular cation. Moreover, Mg is an important cofactor for many enzymes especially those involved in phosphate transfer reactions. Mg is therefore essential in the regulation of the metabolism of other ions and cellular functions. Mg deficiency has been shown to be associated with fatal cardiovascular diseases such as cardiac arrhythmias and coronary heart disease, as well as with risk factors for these diseases, such as hypertension, and diabetes mellitus. Clinical evaluation of Mg status has been limited by the lack of suitable technology for measuring this cation. Although the measurement of serum total Mg is routinely available, ionized Mg is physiologically active. Furthermore, most of the body's Mg is present in the intracellular space. Our findings showed that serum total Mg was similar in all groups, but patients with arrhythmias and diabetes mellitus revealed lower levels of serum ionized Mg. On the other hand, patients with essential hypertension exhibited higher intraerythrocyte Mg concentrations than healthy controls. The measurement of serum total Mg may obscure the diagnosis of an abnormality in Mg metabolism in patients with arrhythmias and diabetes mellitus. Furthermore, the intracellular accumulation of Mg does not support the hypothesis of Mg deficiency in patients with essential hypertension. PMID- 10375960 TI - [Urinary excretion of aquaporin-2 in water metabolism disorders]. AB - Water metabolism plays an essential role in the homeostasis of body fluids in animals and humans. It is regulated by arginine vasopressin (AVP), renal function and water drinking. Disorders of water metabolism result in an increase or decrease in a body water or fluid, which manifest as hyponatremia, hypernatremia, polyuria, dehydration or edema. In the pathogenesis of such pathological conditions AVP is either directly or indirectly involved. Aquaporin-2 (AQP-2) is an AVP-dependent water channel in renal collecting duct cells. Approximately 3% of AQP-2 is excreted into the urine, which is measurable by RIA or Western blot using a specific antibody against AQP-2. There was positive relationship between urinary excretion of AQP-2 (UAQP-2) and plasma AVP levels in normal subjects. UAQP-2 varied in a wide range under ad libitum water drinking. The level of UAQP 2 was one eighth less in patients with central diabetes insipidus than in normal subjects, and it was 2.8-fold greater in patients with water retention. A hypertonic saline infusion test manifested the difference in the UAQP-2 response to an increase in plasma osmolality between the patients with central diabetes insipidus and the normal subjects. Acute oral water load clarified the impaired water excretion and the persistent elevation of UAQP-2 in patients with water retention. Such increased UAQP-2 was linked to nonsuppressible levels of plasma AVP despite hypoosmolality. These results indicate that UAQP-2 is a useful marker to diagnose disorders of water metabolism. PMID- 10375961 TI - [Hyponatremia and inflammation]. AB - We experienced four cases with hyponatremia due to SIADH, which seems to be related to inflammation. The plasma Na concentration decreased when the patients had fever and increased plasma CRP level. In such conditions, plasma vasopressin concentration (PAVP) and the plasma interleukin-6 (IL-6) concentration were increased. There was significant correlation between them. The animal experiments were carried out to investigate the role of interleukin in the development of SIADH. Intravenous administrations of IL-1 beta increased AVP, atrial natriuretic hormone (ANH) and ACTH. The changes in AVP and ACTH were abolished by the pretreatment with an intravenous administration of indomatacin. Moreover, the intracerebroventricular administration (ICV) of IL-1 beta also increased AVP, atrial natriuretic hormone (ANH) and ACTH. The pretreatment of indomatacin attenuated the changes in AVP and ACTH. The intravenous administration of IL-1 beta increased the urinary sodium excretion. The pretreatement of HS142-1, an ANH antagonist, abolished the increase in urinary sodium excretion induced by IL-1 beta. These results suggested that the interleukin play an important role in the development of SIADH associated with inflammation. PMID- 10375962 TI - [Serum uric acid level and fractional excretion of urate in fluid and electrolyte disturbances]. AB - Changes in renal urate clearance may reflect altered glomerular dynamics more precisely than other commonly used indices. Extracellular fluid volume expansion is known to be correlated with uricosuria, while volume depletion is associated with decreased urate excretion. The diagnostic usefulness of measuring serum urate levels and fractional excretions of urate in SIADH, incipient diabetic nephropathy, and prerenal azotemia have been reviewed. In SIADH, profound hypouricemia and markedly increased fractional excretion of urate (FEUA) accompany the extracellular fluid volume expansion. In prerenal azotemia, decreased FEUA may represent a reliable indicator of prerenal azotemia in the differential diagnosis of acute renal failure. In incipient diabetic nephropathy, glomerular hyperfiltration may increase renal urate clearance and lower the serum uric acid level. Hypouricemia may also predict the future progression of incipient nephropathy in type II diabetes. PMID- 10375963 TI - Gs alpha knockouts in mice and man. PMID- 10375964 TI - [Suggestions and propositions to resolve some issues for standardization of prothrombin time and activated partial thromboplastin time]. AB - Prothrombin time (PT) and activated partial thromboplastin time (APTT), popularized as a routine assay for screening blood coagulation disorders and monitoring anticoagulant therapy, still involve some issues regarding standardization. In this lecture, we present propositions to resolve these problems in respective laboratory. Although international normalized ratio (INR) calculated by international sensitivity index (ISI) of PT reagent seems to improve discrepancy of sensitivity between reagents, local calibration of sensitivity of PT reagent in respective laboratories (local SI) is reasonable to make INR/ISI system more useful. However, local calibration of reagent is not easy by WHO recommended method in a small size laboratory. By using AK calibrant (IMMUNO AG), one of calibration plasma for INR, we investigated its possibility to calibrate local SI in four different reagents, compared with the recommended methodology. The results led the following process to determine reagent and calibrate local SI for practical use of INR/ISI system. (a) Use PT reagent of which ISI is close to 1.0 if possible, and utilize manufacture's ISI as is for INR. (b) Select PT reagent labeled specific ISI for an instrument as the same as used in the lab., and use the manufacture's ISI as is, if impossible to choose small ISI reagent. (c) If use a reagent of which ISI is close to 2.0 and shown no specific ISI for used detector, adjustment of local SI by commercial calibration plasma is recommended when unavailable warfarinized patient plasma. In APTT, we attempted to evaluate sensitivity between five different APTT reagents with a patient model by hemophilia A plasma contained various FVIII: C. This model reflected difference of sensitivity between reagents in results. Because standardization of APTT is not improved in this point, certification of APTT pattern in each laboratories with patient models is required for not only monitoring of heparinization, but also screening of typical coagulation disorders such as hemophilia and von Willebrand disease. PMID- 10375966 TI - [Detection of activated platelets by flow cytometry]. AB - While bleeding time and platelet aggregometry are the two standard clinical tests of platelet function, an increasing number of flow cytometric studies using activation-dependent monoclonal antibodies have been reported recently. Those include an IgM antibody, PAC-1, which binds to the fibrinogen binding site exposed by a conformational change in the GPIIb-IIIa complex of activated platelets. In our study, platelets in whole blood bound to PAC-1 when activated by a wide range of ADP concentrations. ADP-induced PAC-1 binding to platelets was suppressed in individuals who had taken an oral antiplatelet drug, ticlopidine. Whole blood flow cytometry have advantages such as minimal artifact and smaller sample volume. Platelet activation and its suppression by antiplatelet therapy may be evaluated by using ADP-induced PAC-1 binding in many clinical settings where platelet hyperreactivity and/or circulating activated platelets play a role. PMID- 10375965 TI - [Cell cycle-dependent change of the adhesive character of CD34+ progenitor cells and their VLA-4 expression]. AB - We identified the cell cycle status of CD34+ cells of steady-state bone marrow (BM) and peripheral blood (PB) obtained from healthy volunteers, and those of BM and apheresis PB samples collected from donors who had been administered granulocyte colony-stimulating factor (G-CSF). Regardless of whether G-CSF treatment was undergone, more than 10% of CD34+ cells in the BM was in the S + G2/M phase. In contrast, less than 2% of CD34+ cells in the PB was cycling. After co-culturing BM CD34+ cells with a monolayer of the stromal cell line MS-5 for 1 hour, some cells adhered to the stroma. The percentage of cells in the S + G2/M phase among these adherent cells was higher than that among the non-adherent cells. Flow cytometric analysis revealed that CD34+ cells in mobilized PB expressed less VLA-4 than those in BM and that in in vitro-cultured non-adherent cells exhibited a lower level of VLA-4 expression than adherent cells. In addition, CD34+ cells in the G0/G1 phase expressed lower levels of VLA-4 than those in the S + G2/M phase. These findings suggested that the reduced expression of adhesion molecules such as VLA-4 by the progenitor cells in the G0/G1 phase of the cell cycle result in the release of progenitor cells from the hematopoietic microenvironment to peripheral blood. PMID- 10375967 TI - [A clinical application of laser scanning cytometer--the significance in the cytology as an additional diagnostic technique]. AB - Laser scanning cytometer (LSC) is a new machine similar to flow cytometer but with advantages for certain clinical and research applications. LSC is a microscope based and measures cells on a slide with the position of each cell on the slide. This new technique of LSC can be utilized on extremely small specimens and enables direct correlation of all of the measured fluorescent parameters with light microscopic cytologic morphology. To date, LSC has been successfully used to perform DNA content analysis of numerous specimen types and automated analysis of fluorescence in situ hybridization specimens. In this report, we describe characteristics of LSC comparison with flow cytometry and a clinical application of LSC focused on DNA content analysis in clinical specimens with pulmonary disorders. LSC provides a number of benefits that may make it more suitable for clinical laboratories than FCM. PMID- 10375968 TI - [Homeostasis of antioxidant status in hemodialysis patients]. AB - Oxidative stress, which occurs when there is excessive free-radical production or low antioxidant levels, makes significant contributions to pathogenesis in many human diseases. Cardiovascular disease is the major cause of mortality in patients receiving hemodialysis. For these patients, oxidative stress and increased lipid peroxidation may contribute to increased risk of atherosclerosis. The aim of this study was to determine if hemodialysis patients were associated with disturbance of homeostasis of antioxidant status. In this experiment, total antioxidant status of serum is measured by its ability to inhibit generation of free radicals from 2,2'-amino-di-[3-ethylbenzthiazole sulphonate] by metmyoglobin and hydrogen peroxide. Status of radical scavengers, such as serum total protein, albumin, uric acid and total bilirubin, was also measured. Blood were collected from three different episodes of hemodialysis. In the first group (n = 29), blood were collected before and after hemodialysis. In the second group (n = 29), blood were collected after dialysis and before next hemodialysis. In the third group (n = 8), blood were collected before hemodialysis. After last hemodialysis, patients started ingesting vitamin C and blood were collected before next hemodialysis. There was a marked reduction of total antioxidant status after hemodialysis in the first group. There was a marked increase in total antioxidant status before next hemodialysis in the second group. High doses of vitamin C caused increase in total antioxidant status in the third group. In conclusion, disturbance of homeostasis of total antioxidant status were observed in patients receiving hemodialysis. This may play a role in the pathogenesis in these groups. PMID- 10375969 TI - [Development of a sensitive enzyme immunoassay for protein 1/Clara cell 10 kDa protein using monoclonal antibodies]. AB - Protein 1 (P1)/Clara cell 10 kDa protein is a dimer of identical subunits of 70 amino acids. Designed to expand its clinical significance, a sensitive sandwich enzyme-linked immunosorbent assay (ELISA) for P1 was developed using two monoclonal antibodies, which were recently generated and characterized in this laboratory. The precision, reproducibility and specificity of the assay were quite satisfactory. Species specificity was also satisfactory. The detection sensitivity was 5 ng/l; high enough to quantify P1 in several body fluids. P1 was distributed in the value of 13 to 25 micrograms/l, 153-4300 micrograms/l and 145 8600 micrograms/l, in serum, bronchoalveolar lavage fluids and seminal fluids, respectively. PMID- 10375970 TI - [Quantitation and heterogeneity of serum alpha 2-macroglobulin]. AB - alpha 2-Macroglobulin (alpha 2M) is a major protease inhibitor in human plasma. In this study, serum alpha 2M was determined by means of immunoelectrophoresis (IEP), rocket immunoelectrophoresis (R-IEP), laser nephelometry (LN) and single radial immunodiffusion (SRID) in 133 subjects. The values obtained by these methods coincided well in most specimens, but some specimens showed disagreement. The sera of alpha 2M deficiency found by IEP were analyzed by the other methods. We found that R-IEP gave the results of alpha 2M deficiency most frequently while the other methods gave the results of normal alpha 2M level for some specimens. We studied on the presumption that the phenomenon is due to heterogeneity in molecular weight of alpha 2M. After alpha 2M was purified by affinity chromatography and gel filtration chromatography, we further employed gel filtration on a high-performance liquid chromatography (HPLC) to isolate alpha 2M with different molecular size. Two peaks of immunoreactive alpha 2M were found by the gel-filtration HPLC. When we performed IEP for both portions, the first portion with higher molecular size showed larger rate of migration than the second portion. Serum alpha 2M quantitation with SRID is not appropriate because molecular weight influences on the determination by SRID. We conclude that R-IEP is best suitable for the detection of decrease of serum alpha 2M, although this method requires expertness of manipulation. PMID- 10375971 TI - [Dibucaine number (DN) and fluoride number (FN) of L330 I mutant recombinant cholinesterase by use of various substrates]. AB - Dibucaine number (DN) and fluoride number (FN) of the recombinant 330 I mutant ChE (r330 I) expressed in human kidney cells (293 cell) were compared with recombinant usual ChE (rUU), by several assay kits and substrates. All of them showed lower the values compared with rUU. However, the r330 I/rUU ratios about FN determined by several substrates were higher than that determined by propionyl thiocholoneiodide (PTCI), which was recommended by American Association for Clinical Chemistry. In conclusion, commercially available assay kits may not be suitable for the determination of L330 I. PMID- 10375972 TI - [The 43rd Congress of the Japan Rheumatism Association. Sapporo, Japan. June 3-5, 1999. Abstracts]. PMID- 10375973 TI - [Patient perceptions and satisfaction of psychiatric services at their discharge]. AB - We conducted a multi-central survey on patient's perception and satisfaction of 31 psychiatric services at their discharge. The subjects were 364 patients who agreed to participate in the survey among 471 discharged patients between August 10th and September 10th, 1997. We excluded 48 patients emergently transferred to another hospitals, 47 patients with dementia or mental retardation, and 12 patients who refused to participate. Of the subjects, 326 patients (89.6%) responded. The rates of patients who rated the psychiatric care positively ranged between 79.4% (Amenity) and 93.6% (Overall satisfaction). Older patients tended to be more satisfied with psychiatric care than younger patients. Patients with schizophrenia and mood disorders rated the psychiatric care more positively, whereas patients with personality disorders rated negatively. Patients with neurosis rated the care positively in informed consent, but negatively in other items. Patients with the 60's score in Global Assessment of Functioning Scale rated higher in nurses and clerks than patients with other scores. Patients who have received their care in non-voluntary admission rated significantly lower in informed consent than other patients. There were no significant correlation of patient satisfaction with former admission and type of ward. Our results indicate that patient satisfaction survey could contribute to improve psychiatric care regarding patient characteristics in Japan. PMID- 10375974 TI - [Criticisms on Yasuo Okada's article: on psychiatric terminology]. PMID- 10375975 TI - [Development and use of the Japanese version of the self-report Yale-Brown Obsessive Compulsive Scale]. PMID- 10375976 TI - [(EPIC XI) proceedings (recent advances in event-related brain potential research)]. PMID- 10375977 TI - [ERPs changes during neuroleptic treatment in schizophrenia--a vulnerability marker in schizophrenia]. AB - P300 amplitude reduction and P300 latency prolongation are consistent findings in schizophrenia, but it is unclear if these abnormalities were the effect of current or past neuroleptic treatment or were present at the onset of illness. We previously recorded ERPs in drug free schizophrenic patients (45 neuroleptic naive and 56 previously treated with neuroleptics). In that study, P300 amplitude reduction was observed in both the neuroleptic-naive and the previously treated patients. However, both N200 and P300 latencies were prolonged only in the previously treated schizophrenic patients. In this study, we investigated ERPs in 60 drug free schizophrenic patients before and after neuroleptic treatment was begun. According to DSM-IV, schizophrenia subtype classification, 26 cases were paranoid type, 14 were disorganized, 2 catatonic and 18 undifferentiated. Twenty six of the patients were neuroleptic-naive and 34 had been previously treated. Sixty gender- and age-matched healthy controls were also investigated. ERPs were recorded during an auditory oddball task. The scalp EEGs were recorded from AgAgCl electrodes at 16 sites according to the international 10-20 system. Clinical symptoms were assessed using the Brief Psychiatric Rating Scale (BPRS). Before treatment, all schizophrenic patients displayed larger N200 amplitudes than the controls; however, increases in N200 amplitudes were not observed after neuroleptic treatment was begun. Both N200 and P300 latencies in the patients before treatment were prolonged only in those previously treated. Neuroleptic naive patients demonstrated prolongation of both N200 and P300 latencies only after treatment. P300 amplitudes in patients were increased by neuroleptic treatment; but patients had smaller P300 amplitudes than the controls even after treatment. The change in P300 amplitudes (Pz) and the change in total BPRS scores by neuroleptic treatment were positively correlated in the patients whose duration of illness was six months or less (mean: 2.4 months). However, no correlation was observed for patients whose duration of illness was over six months (mean: 49.7 months). There were no significant differences in ERPs changes among subtypes. These results suggested that the P300 amplitude should be considered a vulnerability marker in schizophrenia and that both N200 and P300 latencies might be markers for neuroleptic exposure. PMID- 10375978 TI - [An analysis of public psychiatric hospital activities]. PMID- 10375979 TI - [A psychopathological study of aggression in schizophrenic patients]. AB - For some schizophrenic patients violent behavior occurs during acute phases of their illness. Some of these patients who exhibit violent behavior recover quickly from this phase. Further, there is a marked improvement in the quality life during remission. In this study the psychopathological features of aggression in 24 schizophrenic patients were considered. All of these patients were involuntary emergency admissions due to an uncontrollable violent episode. The interaction between aggression and delusion was categorized as follows: convergent type, aggression due to delusion; reactive type, aggression instigated by hallucination or delusional perception; and divergent type, random outbursts of violence and aggression. The intensity of catatonic features and fixed targets of violence distinguishes convergent type violent schizophrenic patients from reactive and divergent type patients. Convergent aggressive behavior can be more easily treated then the other two types and should have significant bearing over the course of treatment. PMID- 10375980 TI - [Differential diagnosis of schizophrenic symptoms complicated with brain anomalies including bilateral temporal arachnoid cysts by eye mark recorder and PET: a case study]. AB - Bilateral temporal arachnoid cysts and other intracranial congenital lesions including a moderately large left temporal arachnoid cyst accompanied by remarkable dysplasia of the temporal lobe in particular were discovered by chance during computerized axial tomography of a 26-year-old Japanese male who had been diagnosed as schizophrenia approximately 10 years earlier. A detailed re assessment revealed no other organic symptoms or signs. His symptoms and clinical course met the DSM-IV criteria for schizophrenia, disorganized type. Based on his symptoms, positron emission tomography (PET) and the eye-movement recording test developed by Kojima et al. were performed. In addition, psychological tests including WAIS, Rorschach Test, and Wechsler's Memory Test were administered for further differential diagnosis. PET using continuous inhalation of oxygen 15-gas revealed a regional decrease in CBF and CMRO2 in the superior medial frontal lobe including the anterior cingulate gylus, findings sometimes associated with schizophrenia. However, no abnormal findings were noted around the arachnoid cysts. In the eye-movement recording test, several parameters including the responsive search score (RSS) were about the same level as that commonly observed in schizophrenics and are classified as schizophrenia by discrimination analysis. The psychological tests offered no reason to doubt the diagnosis of schizophrenia. Thus, the patient was diagnosed as schizophrenia with arachnoid cysts and other intracranial lesions. The way of diagnosis we used here might bring forth a breakthrough in schizophrenia research by differentiating schizophrenia from the other organic brain diseases. PMID- 10375981 TI - [Actual state of involuntary admission by the prefectural governor--a research report on the involuntary admission in 1995. Committee on Psychiatric Services and Law. Sub-Committee on Measures of Security and Justice]. PMID- 10375982 TI - Jamaican doctors can help to prevent interpersonal violence. PMID- 10375983 TI - Sir Harry Annamunthodo. PMID- 10375984 TI - A behaviour risk factor survey in Jamaica. AB - A population based probability sample of 958 persons (454 males and 504 females) aged 15 to 49 years was surveyed in Jamaica in late 1993 for lifestyle and behaviour risk factors. Demographic characteristics of the sample were comparable to the general population, 60% of persons visited a private doctor the last time that they were ill. Based on self-reporting, 18% of the women and 8% of the men were hypertensive and 4.8% of the women and 3.3% of the men were diabetic. 26% of the men and 8% of the women had never had their blood pressure taken. 40% of the women had never had a Papanicolaou smear, 29% had never had a breast examination and 33% said that they were overweight compared with 18% of men. Smoking cigarettes and marijuana was more common among men (36%) than women (11%), as were drinking alcohol (79% of men, 41% of women) and heavy alcohol use (30% of men, 9% of women). Injuries requiring medical attention in the previous five years were reported by 40% of the men and 15% of the women. 34% of the men and 12% of the women regularly carried a weapon and 18% of the sample had participated in or witnessed at least one violent act in the previous month. Most of the people interviewed used a contraceptive method; 10% were not sexually active. Significantly more men than women had two or more sexual partners in the previous year (54% vs 17%, p < 0.001) or reported ever having a sexually transmitted disease (29% vs 9%, p < 0.001). Younger persons were more sexually active and more likely to use condoms during their most recent sexual intercourse. Higher socio-economic status and educational level generally had a more positive effect on health behaviour. This survey provides vital information relevant to planning health promotion campaigns and assessing their success. PMID- 10375985 TI - Clustering of cases of Mycobacterium fortuitum infection investigated by molecular typing. AB - Organisms of the Mycobacterium fortuitum complex are recognised but uncommon causes of pulmonary disease, primary cutaneous disease and a wide spectrum of nosocomial infections. M fortuitum was isolated from 20 patients over a 15 month period, with an apparent clustering of isolates occurring from January to March 1994. The molecular epidemiology of this clustering was investigated using an arbitrary primer polymerase chain reaction method (AP-PCR). 21 isolates were studied, which yielded 13 distinct profiles. Multiple isolates from a single patient yielded identical profiles. All of seven isolates recovered during the six week period from January to March 1994 shared a common profile which was distinct from all other isolates, suggesting that a single strain was isolated from specimens from all seven patients. The source of this cluster is uncertain. We can find no epidemiological basis for an episode of cross-infection within the hospital environment, and it is assumed that contamination of the specimens during collection, transport or processing was responsible for the "pseudo outbreak" of M fortuitum. PMID- 10375986 TI - Methicillin resistant Staphylococcus aureus. AB - The prevalence of methicillin resistant Staphylococcus aureus (MRSA) at the General Hospital, Port-of-Spain, between June 1995 and May 1996 was determined. The MRSA prevalence rate was 4.6% of all S aureus isolates, with all but one nosocomially acquired. 15 isolates were associated with infections, and three were colonizing strains. 17 of the 18 patients with MRSA had received antibiotics previously, including 13 who had received multiple antibiotics. Skin and soft tissue were the sites of infection and colonization in 12 cases; and surgical wards and the Intensive Care Unit (ICU) accounted for 16 MRSA isolates. All isolates were sensitive to vancomycin and all but one were resistant to gentamicin. MRSA occurred sporadically in a wide distribution of wards and physicians' services, although the isolation of three strains from the ICU and three strains from a surgical ward were temporally related. Only one of two deaths was attributable to MRSA. Control of the spread of MRSA in this hospital must include the reinforcement of the appropriate use of antibiotics, hand washing and appropriate isolation of patients in the surgical and intensive care wards. PMID- 10375987 TI - Neuroprotection by caffeine and pentoxifylline during experimental cerebral ischaemia. AB - Cerebral ischaemia was induced in anaesthetized rats by occlusion of the ipsilateral common carotid and middle cerebral arteries. The response to ischaemia was assessed by the reduction of the amplitude of recorded somatosensory evoked potentials (SSEPs), and the rate of recovery of the SSEPs during reperfusion. Caffeine and pentoxifylline when applied at 70 mM to the cortex for 60 min prior to induction of ischaemia significantly reduced the ischaemia induced attenuation of the SSEPs and hastened recovery to control levels. In contrast, application of normal saline or of the drugs for 15 min did not reduce the effect of ischaemia on the SSEPs. These results suggest that caffeine and pentoxifylline have potential roles in the management of patients with cerebral ischaemia. PMID- 10375988 TI - Necrotising enterocolitis: a management protocol for developing countries. AB - 28 cases of necrotising enterocolitis (NEC) comprising 11 term and 17 preterm patients were diagnosed between January 1990 and December 1995 at the University Hospital of the West Indies (UHWI). Treatment was in accordance with a management protocol which emphasised aggressive screening of potential cases, early laparotomy for bowel perforation and primary anastomosis after small bowel resection. There were three deaths among the 13 cases of bowel perforation. Centres in developing countries can achieve rates of survival comparable to those in the developed world in babies with NEC weighing over 1000 grams by adopting the UHWI management protocol. PMID- 10375989 TI - Medical admissions of oncology patients to the University Hospital of the West Indies. AB - A prospective study of 80 oncology patients (42 men, 38 women; mean age 50.3 years) admitted to the University Hospital of the West Indies, Jamaica, was conducted over a six month period (August 1, 1995 to January 31, 1996). There were 103 admissions representing 8.7% of total admissions to the medical wards. Solid tumours and haematological malignancies accounted for equal proportions of admissions. 62% were emergency admissions. Investigation of constitutional symptoms, abnormal physical findings, infection and chemotherapy were the commonest reasons for admission. Complications developed in 42.7% of admissions, the commonest being renal and/or hepatic impairment; anaemia, leukopaenia and thrombocytopenia; and nosocomial infections. 35% of the patients died during the study period. The mean length of stay was 12.9 days (SD 12.8). Mean hospital stay was significantly longer in admissions involving an initial diagnosis of cancer and in those resulting in complications (p < 0.001). PMID- 10375990 TI - Magnesium sulphate for eclamptic seizures? PMID- 10375991 TI - Granulosa-theca cell tumour of the ovaries. A late metastasizing tumour. AB - Granulosa-theca cell tumours are ovarian neoplasms of low malignancy with hormone secreting potential, accounting for 2-3% of all ovarian cancers. They have an uncertain clinical course and a potential for late recurrence after surgical removal. Clinical features of a patient presenting with pulmonary metastases 21 years after removal of the primary tumour are described, along with a review of the management options. PMID- 10375992 TI - The Guillain-Barre syndrome following dengue fever. AB - A 44 year old female presented with fever, muscle aches, rash and a low platelet count. IgM antibody to dengue virus was positive. Two weeks later she developed a flaccid areflexic quadriparesis. Nerve conduction studies showed a predominantly demyelitinating sensory motor polyneuropathy consistent with Guillain-Barre syndrome. Despite the relatively common occurrence of dengue fever, an associated polyradiculoneuropathy is distinctly uncommon. PMID- 10375993 TI - Malaria in the Americas. PMID- 10375994 TI - [Efficient synthesis of nucleosides labeled with stable isotopes and their application to structural biology]. AB - 1) A series of synthetic works on nucleosides appropriately labeled with stable isotopes of deuterium, carbon-13, and nitrogen-15 has been undertaken, confronting the strong demands for the nucleosides in the NMR spectroscopic study deeply related to structural biology, and the synthetic methods of (2'R)- and (2'S)-2'-deoxy[2'-2H]ribonucleosides, 2'-deoxy[5'-2H]ribonucleosides, [5' 13C]ribonucleosides, 2'-deoxy[5'-13C]ribonucleosides, 2'-deoxy[4-15N]cytidine, [4 15N]cytidine, 2'-deoxy[6-15N]adenosine, and [6-15N]adenosine were developed more efficiently than ever; some oligodeoxyribonucleotides were constructed by the use of these materials and found to be extraordinarily feasible for the NMR spectroscopic studies. 2) A novel approach to oligonucleotide synthesis on CPG has been established by the use of a base-labile protecting group, i.e., 2 (levulinyloxymethyl)-5-nitrobenzoyl (LMNBz) protecting group for the 5'-hydroxyl group of nucleoside 3'-phosphoramidites. PMID- 10375995 TI - [Terpenoids and steroids from several euphorbiaceae and pinaceae plants]. AB - During the course of a search for biologically active constituents from unexamined plant sources, several biogenetically interesting new di- and tri terpenes and steroids were isolated from several weeds and shrubs of Euphorbiaceae and the bark, leaves and cones of several Pinaceae trees which had been treated as wastes in the forestry industry. Euphorbia supina contained 3,4 seco-5 alpha- and 5 beta-adian-4(23)-ene-3,5-diols and related oxides, oxygenated fern-8-en-3 beta-ols named supinenolones A-E and unusually migrated oxyfernanes having (9S)- and (9R)-7(8-->9)abeo-9-D:C-friedo-B':A'-neogammacerane skeletons named spirosupinane and neospirosupinane, while E. chamaesyce contained 3,4-seco oleana-4(23), 18-dien-3-oic acid, 3,4-seco-8 beta H-ferna-4(23),9(11)-dien-3-oic acid and two oxygenated obtusifoliols. The bark of Phyllanthus flexuosus (Euphorbiaceae) contained 11 beta-hydroxy-D:A-friedo-olean-1-en-3-one, lup-20(29) ene-3 beta, 15 alpha-diol, olean-12-ene-3 beta,15 alpha-diol and olean-12-ene-3 beta,15 alpha,24-triol together with trichadenic acid B for which we revised the structure to 3 beta-hydroxy-D:A-friedo-oleanan-27-oic acid. Two 26-nor-D:A-friedo olean-14-enes were isolated from P. watsonii. Regarding Pinaceae trees, an unusually migrated abieslactone [(3R, 7S, 9R, 23R)-7-hydroxy-3-methoxy-8-oxo-7(8- >9)abeo-lanost-24-eno-26,23-lactone], named spiroveitchionolide, was isolated from the bark of Abies species, besides nine abieslactone analogues. Two pairs of unusually migrated serratanes, piceanonols A and B and jezananals A and B having novel skeletal systems of 14(13-->12) abeo- and 16(15-->14) abeo-serratanes named piceanane and jezanane, respectively, were also isolated from the stem bark of Picea species, besides three 14 beta,15 beta-epoxyserratanes and two 13 alpha,14 alpha-epoxyserratanes. The leaves of Larix kaempferi contained two deformed abietanes named karamatsuic acid (9,10-seco-9,10 alpha-epoxyabieta-8,11,13-trien 18-oic acid) and larikaempferic acid [9 alpha,13 alpha-epoxy-8-oxo-9(8-->7)abeo-7 beta-abietan-18-oic acid], as well as the cones to contain 8 alpha,12 alpha epidioxy-15-hydroxyabiet-13-en-18-oic acid, three diepoxy-abietan-18-oic acids and two new dehydroabietic acid analogues. Several of the above compounds exhibited inhibitory effects against tumor-promoting and DNA topoisomerase II activities. PMID- 10375996 TI - [Studies on the alkaloids of Erythrina plants]. AB - The alkaloidal components of eight Erythrina plants (Leguminosae), E. arborescence Roxb., E. orientalis (L.) Murr, E. crysta-galli Linn, E. crysta galli (L.) cv. Maruba Deiko H. Murata, E. x bidwilli Lindl, E. poeppigiana (Walp) O. F. Cook, E. glauca Willd, and E. variegata L. were examined. As a result of this study, five new oxo-erythrinan alkaloids, erythrinine (8), 11 hydroxyerysotrine (9), erysotramidine (10), erytharbine (11), crystamidine (12) and a di-benz[d,f]azonine type alkaloid, erybidine (2), were isolated respectively. Two tetrahydroprotoberberine type alkaloids, scoulerine (4) and coreximine (5), were also isolated from E. orientalis (L.) Murr. A new synthetic route to erythrinan alkaloids was developed, via the cis -C/D-ring fused 15 methoxy-16-hydroxyerythrinan-2, 8-dione (49) as a key intermediate, from the enol methyl derivative (48) which was obtained by Birch reduction of the benzyl amide (47). The total synthesis of (+/-)-erysotramidine (10), an oxo-erythrinan alkaloid, including a novel ring cleavage of the aza-tricyclo[3.2.0.0] compound (70) with phenylselenyl chloride is described. PMID- 10375997 TI - [Development and applications of new reactions for construction of basic structures of natural products]. AB - This review deals with the development of efficient methods to construct the basic structure of natural products and the versatile methods to control the stereochemistry, and these methods were applied to the synthesis of natural compounds. The photochemical spirodienone formation reaction was applied to the synthesis of proaporphine alkaloids. The alternative spirodienone formation reactions by the metal-catalyzed degradation reaction of phenolic alpha diazoketones were applied to many natural spirocyclic compounds, such as chamigrane type sesquiterpenes, spirovetivane type phytoalexins, marine natural products, and so on. Lewis acid mediated spirocyclization reaction of cyclohexene bis-acetal derivative was developed, and this reaction was applied to the synthesis of aphidicolane and stemodane diterpenes. The regioselective cleavage reaction of the cyclopropane ring of tricyclooctanone derivatives was used for the syntheses of diquinane and triquinane compounds. A chiral pool synthesis of several aromadendrane sesquiterpenes was achieved via common tricyclic enone intermediates. The synthesis of macrocarpals, coupling products of aromadendrane skeleton and isopentylphloroglucinol dialdehyde, was also accomplished for the first time using an arene Cr(CO)3 complex as a chiral benzyl cation equivalent. The Fe(diene)(CO)3 complexes were used for the highly stereoselective asymmetric synthesis of several natural products, such as insect pheromones and alkaloid, as a versatile mobile chiral auxiliary. PMID- 10375998 TI - [Thromboxane A2 antagonist--discovery of seratrodast]. AB - We were interested in RCS (rabbit aorta contracting substance) and SRS-A (slow reacting substance of anaphylaxis) and their involvement in human bronchial asthma. When we started our anti-asthmatic drug research in the 1970's. We synthesized a lot of chemical compounds and eventually discovered that AA-861 inhibited the generation of SRS-A from the lung tissue of actively sensitized guinea pigs. AA-861 was found to be a potent 5-lipoxygenase inhibitor. This compound reduced experimental allergic asthma in guinea pigs, but it is easily metabolized in the body. More recently, we found a novel compound, AA-2414 (seratrodast), which is not metabolized in the body. AA-2414 proved to be not a 5 lipoxygenase inhibitor, but a thromboxane A2 (TXA2) receptor antagonist. Seratrodast is the first receptor antagonist that is being developed as an anti asthmatic drug. Seratrodast inhibits both immediate-, late asthmatic responses in guinea pigs, and also reduces airway hyperresponsiveness in dogs. The anti asthmatic action of seratrodast in animal models indicates that the drug should be of use in the treatment of human asthmatics. In clinical studies, seratrodast showed a marked effect to improve clinical parameters in bronchial asthma. It is also reported that seratrodast is free from harmful aftereffects. Clinical trials are under way in the US. PMID- 10375999 TI - [Investigation on the marker substances of crude drugs in formulations. I. Marker substances for the identification of astragali radix in kampo and drinkable preparations]. AB - There are few reports about marker substances for the identification of Astragali Radix in formulations. First, constituents analysis was performed by HPLC for the screening of a marker substance, using several lots of Astragali Radix and its fluid extracts. As a result of the analysis, one of the main components was clearly detected even in the fluid extracts, and thought to be a good marker substance. This component was identified to be calycosin (7,3'-dihydroxy-4' methoxyisoflavone) by the structural analysis. Then, the identification methods of Astragali Radix and its fluid extracts in formulations were investigated with an Astragali Radix extract as a reference standard solution and calycosin as a marker substance. This method was applied to three different kinds of Kampo formulations and three different kinds of drinkable preparations. Consequently, calycosin was clearly detected by a HPLC-multi wavelength detector system, in all investigated formulations. Calycosin will be successfully used as a marker substance for the identification of Astragali Radix and its fluid extracts in formulations. PMID- 10376000 TI - [Relationship between biological activity and heparin-affinity sites of recombinant human basic fibroblast growth factor CS23 mutein]. AB - A recombinant human basic fibroblast growth factor CS23 mutein (rhbFGF-CS23) obtained from Escherichia coli cells has proliferation-stimulating activity for a fetal bovine heart endothelial cell line, ATCC CRL 1395 (biological activity), and strong affinity for heparin (heparin-affinity) similarly to the natural human basic fibroblast growth factor. Plural species having different kinds of heparin affinity were formed by acetylation of rhbFGF-CS23 with acetic anhydride. To clarify the relationship between the sites with heparin-affinity and the biologically active sites, we have investigated the acetylation sites by peptide mapping and the biological activity of the acetylated species. Consequently, the sites with heparin-affinity in rhbFGF-CS23 are found to be Lys26, Lys119, Lys125, Lys129, and Lys135 in the primary structure, and these sites with heparin affinity except Lys26 are also clarified to be very important to retain the biological activity of the factor. PMID- 10376001 TI - [Examination of the stability of chloral hydrate and its preparation by capillary electrophoresis]. AB - Stabilities of chloral hydrate in an aqueous solution and its medicated syrup were examined by high performance capillary electrophoresis. Analysis of the concentration of chloral hydrate indicated that there was no obvious change in the concentration of chloral hydrate both in the aqueous solution and in the syrup preparation after keeping them for 3 months at room temperature or at 60 degrees C. The lowering of pH was more obvious in the syrup solution than in the aqueous solution, and this tendency was estimated to be due to the formation of hydrochloric acid. We propose that the stabilities of the preparation of chloral hydrate should be monitored by observing pH changes. PMID- 10376002 TI - [Characterization of the facilitatory modulation of adrenergic neurotransmission via prejunctional purinoceptors in rabbit ear artery]. AB - The purinergic modulation of the release of norepinephrine (NE) from sympathetic nerves in the rabbit ear artery was investigated. Methoxamine, an alpha 1 adrenoceptor agonist, enhanced the NE-release by electrical stimulation (ES) and released large amounts of adenyl purines (ATP, ADP, AMP and adenosine) from the endothelium. Both actions of methoxamine were blocked by prazosin. In addition, the enhancement of the NE-release by methoxamine was abolished by 8-sulfophenyl theophylline (8SPT), a P1-Purinoceptor antagonist. These findings indicated that the endogenous purines and purinoceptors participate in the facilitation of NE release by alpha 1-adrenoceptor stimulation. P1-Purinoceptor agonists, such as adenosine and 2-chloroadenosine, and P2-purinoceptor agonists, such as ATP and beta,gamma-methylene ATP (beta gamma-mATP), enhanced the ES-evoked NE-release. This enhancement was antagonized not only by the P1-purinoceptor antagonist, 8SPT, but also by the P2-purinoceptor desensitizing agent, alpha,beta-methylene ATP. A phosphodiesterase inhibitor, Ro20-1724 potentiates the enhancement of NE release by beta gamma-mATP. On the other hand, an adenylate cyclase inhibitor, SQ22536, inhibited the enhancement of NE-release by beta gamma-mATP. These findings suggested that prejunctional facilitatory purinoceptors exist on the adrenergic nerves of the rabbit ear artery. This receptor may be coupled to adenylate cyclase and is different from well-known P1 and P2 purinoceptors. In the rabbit ear artery, adrenergic neurotransmission may be regulated by endogenous ATP and its metabolites via prejunctional facilitatory purinoceptors, which were initiated by alpha 1-adrenoceptor stimulation; i.e. transsynaptic regulation of neurotransmission. PMID- 10376003 TI - [Effects of mouth wash on the removing beclomethasone dipropionate delivered by pressurized aerosol metered-dose inhaler in the mouth]. AB - Effects of mouth wash (mouth rinsing and gargling) on the removal of drug residues in both mouth and pharynx after the use of pressurized aerosol metered dose inhaler (MDI) were studied. The concentration of beclomethasone dipropionate (BM) in mouth wash after a splay of Becotide inhaler was measured by the method using HPLC. The total amount of the removed BM was measured by a sum of the concentrations of BM in 4 or 5 times of mouth washes in the following 4 kinds of methods. In method 1, mouth wash was done with 5 times of water change after a splay of MDI on wetted mouth. In method 2, mouth wash was done with 5 times of change water on dried mouth. In method 3, mouth wash was done with 4 times of change saliva on wetted mouth. In these methods, the actual inhalation of BM was not done. In method 4, mouth wash was done with 5 times of change water after a splay and a inhalation on wetted mouth. The mouth wash procedures removed totally 47.9%, 51.1%, 31.3%, and 33.3% of a splayed amount of BM in each method, respectively. It was required for the removal of 90% of the totally recovered BM to do one time of mouth wash in method 1, two times in method 2, three times in method 3, and two times in method 4, respectively. These data suggest that the mouth wash procedure is shown to have prophylactic benefit for candidiasis induced by steroid delivered by MDI. PMID- 10376004 TI - [Studies on the constituents of Ligustrum species. XIX. Structures of iridoid glucosides from the leaves of Ligustrum lucidum AIT]. AB - Two new iridoid glucosides, named iso-8-epikingiside and 8-demethyl-7 ketologanin, were isolated together with 8-epikingiside, kingiside, ligustroside, 10-hydroxyligustroside, ligustaloside A and ligustaloside B from the leaves of Ligustrum lucidum AIT. (Oleaceae). The stereochemical structures of these new compounds were elucidated on the basis of spectroscopic evidence. PMID- 10376005 TI - [The application of the endotoxin test for globulin and other blood products]. AB - The application of the endotoxin test for globulin and other blood products were investigated by two different limulus amebocyte lysate (LAL) test methods, colorimetric and kinetic turbidimetric methods, using two endotoxin-specific reagents. By the dilution of the blood products in 40 times or more, spiked endotoxin in the products was recovered accurately showing neither inhibition nor enhancement. The definite difference was not shown between the results obtained by the two LAL test methods. According to the method of the endotoxin test described under General Tests of The Japanese Pharmacopeia (thirteenth edition), JPXIII, the maximum valid dilution (MVD) for these products will be calculated to be 40 or more, so it is capable to measure the endotoxin limit for each product. This study indicates that the endotoxin test is applicable to measure the endotoxin content in globulin and other blood products as an alternative method for the rabbit pyrogen test. PMID- 10376006 TI - The menace of the AIDS-tuberculosis combo: any solutions? PMID- 10376007 TI - How do linker histones mediate differential gene expression? AB - In developing Xenopus laevis embryos the multiple-copy oocyte-type 5S RNA genes are progressively shut down. Results presented in three recent articles 1-3 together demonstrate that replacement of the cleavage stage linker histone B4 by somatic H1 leads to chromatosomes positioned directly over these genes and adjacent sequences so as to occlude the binding site for the critical transcription factor TFIIIA. In contrast, on the somatic-type 5S genes the somatic H1 positions chromatosomes about 65 bp further upstream, thereby leaving the TFIIIA binding site exposed and the genes active. The somatic linker histone thus functions as a specific gene repressor. PMID- 10376008 TI - Knockout of REST/NRSF shows that the protein is a potent repressor of neuronally expressed genes in non-neural tissues. AB - The protein repressor element 1 silencing transcription factor/neuron restrictive silencer factor (REST/NRSF) is a negative regulator of neuronal genes that contain a particular DNA sequence, the neuron restrictive silencer element (NRSE). REST is expressed ubiquitously in non-neural tissues but is down regulated in neural precursors and turned off in postmitotic neurons, suggesting that it can act both to prevent extraneural expression of certain genes and to delay the differentiation of neuronal subtypes. In a recent paper, Chen et al.(1) describe the production of a null mutant for REST in mice and the mosaic inactivation of REST function in chicken embryos. Knockout of REST led to malformations in several non-neural tissues, as well as apoptosis and embryonic lethality in mice. In addition, the expression of several REST target genes was derepressed in non-neural tissues and in neural progenitors in both mouse and chicken embryos. These studies clearly demonstrate that active repression of tissue-specific genes is required for proper tissue differentiation during embryonic development. PMID- 10376009 TI - The genome of Rickettsia prowazekii and some thoughts on the origin of mitochondria and hydrogenosomes. AB - The sequence of an alpha-proteobacterial genome, that of Rickettsia prowazekii, is a substantial advance in microbial and evolutionary biology. The genome of this obligately aerobic intracellular parasite is small and is apparently still undergoing reduction, reflecting gene losses attributable to its intracellular parasitic lifestyle. Evolutionary analyses of proteins encoded in the genome contain the strongest phylogenetic evidence to date for the view that mitochondria descend from alpha-proteobacteria. Although both Rickettsia and mitochondrial genomes are highly reduced, it appears that genome reduction in these lineages has occurred independently. Rickettsia's genome encodes an ATP generating machinery that is strikingly similar to that of aerobic mitochondria. But it does not encode homologues for the ATP-producing pathways of anaerobic mitochondria or hydrogenosomes, leaving an important issue regarding the origin and nature of the ancestral mitochondrial symbiont unresolved. PMID- 10376010 TI - Can't get no SMADisfaction: Smad proteins as positive and negative regulators of TGF-beta family signals. AB - The identification of Smad proteins as molecular components of the transforming growth factor-beta (TGF-beta) signaling cascade has enhanced our understanding of how ligand-mediated activation of TGF-beta receptors leads to modulation of target gene transcription. Recent studies have identified a distinct, structurally related class of Smads which inhibits, rather than transduces, TGF beta family signals. The molecular mechanism of action and the exact signaling pathways that are targeted by antagonistic Smads are not completely understood. These proteins appear to participate in autoregulatory negative feedback loops in which signaling initiated by specific TGF-beta family ligands induces the expression of an inhibitory Smad that then functions to modulate the amplitude or duration of signaling. Negative feedback circuits such as these play important roles in fine-tuning the activity of multifunctional signaling molecules during embryonic patterning and in response to pathologic stimuli in adults. PMID- 10376011 TI - Eukaryotic DNA methylation as an evolutionary device. AB - DNA methylation is catalyzed by a family of conserved DNA methyltransferases and is widespread among protists, plants, fungi and animals. It is however absent in some species and its genomic distribution varies among organisms. Sequence comparisons suggest that known and putative eukaryotic DNA methyltransferases fall into at least five structurally distinct subfamilies. Furthermore, it is now clear that DNA methylation can be involved in several functions, some of which may coexist within the same organism. It can inhibit transcription initiation, arrest transcript elongation, act as an imprinting signal, and suppress homologous recombination. On the basis of these observations, we argue that DNA methylation has been conserved during evolution because it provides unique possibilities for setting up functions of various types. PMID- 10376012 TI - Protein glycosylation in development and disease. AB - N- and O-linked glycan structures of cell surface and secreted glycoproteins serve a variety of functions related to cell-cell communication in systems affecting development and disease. The more sophisticated N-glycan biosynthesis pathway of metazoans diverges from that of yeast with the appearance of the medial-Golgi beta-N-acetylglucosaminyltransferases (GlcNAc-Ts). Tissue-specific regulation of medial- and trans-Golgi glycosyltransferases contribute structural diversity to glycoproteins in metazoans, and this can affect their molecular properties including localization, half-life, and biological activity. Null mutations in glycosyltransferase genes positioned later in the biosynthetic pathway disrupt expression of smaller subsets of glycan structures and are progressively milder in phenotype. In this review, we examine data on targeted mutations affecting glycosylation in mice and congenital mutations in man, with a view to understanding the molecular functions of glycan structures as modulators of glycoprotein activity. Finally, pathology associated with the expression of GlcNAc-Ts in cancer and diabetes-induced cardiac hypertrophy suggest that inhibitors of these enzymes may have therapeutic value. PMID- 10376013 TI - Signaling pathways in phagocytosis. AB - Phagocytosis is an uptake of large particles governed by the actin-based cytoskeleton. Binding of particles to specific cell surface receptors is the first step of phagocytosis. In higher Eucaryota, the receptors able to mediate phagocytosis are expressed almost exclusively in macrophages, neutrophils, and monocytes, conferring immunodefence properties to these cells. Receptor clustering is thought to occur upon particle binding, that in turn generates a phagocytic signal. Several pathways of phagocytic signal transduction have been identified, including the activation of tyrosine kinases and (or) serine/threonine kinase C in pivotal roles. Kinase activation leads to phosphorylation of the receptors and other proteins, recruited at the sites of phagocytosis. Monomeric GTPases of the Rho and ARF families are likely to be engaged downstream of activated receptors. The GTPases, in cooperation with phosphatidylinositol 4-phosphate 5-kinase and phosphatidylinositol 3-kinase lipid modifying enzymes, can modulate locally the assembly of the submembranous actin filament system leading to particle internalization. PMID- 10376014 TI - Generation of evolutionary novelty by functional shift. AB - That biological features may change their function during evolution has long been recognized. Particularly, the acquisition of new functions by molecules involved in developmental pathways is suspected to cause important morphologic novelties. However, the current terminology describing functional changes during evolution (co-option or recruitment) fails to recognize important biologic distinctions between diverse evolutionary routes involving functional shifts. The main goal of our work is to stress the importance of an apparently trivial distinction: Whether or not the element that adopts a new function (anything from a morphologic structure to a protein domain) is a single or a duplicated element. We propose that natural selection must act in a radically different way, depending on the historic succession of co-option and duplication events; that is, co-option may provide the selective pressure for a subsequent gene duplication or could be a stabilizing factor that helps maintain redundancy after gene duplication. We review the evidence available on functional changes, focusing whenever possible on developmental molecules, and we propose a conceptual framework for the study of functional shifts during evolution with a level of resolution appropriate to the power of our current methodologies. PMID- 10376015 TI - Are there molecules of nucleoprotamine? AB - A short critical review of the data related to protamine and nucleoprotamine (DNP) structure is given. A new model is proposed for DNP structure in which protamine molecules are located in channels between the DNA molecules. DNA molecules are arranged hexagonally in the x-y plane, whereas their relative positions with respect to the z-axis are shifted by 0, 1/3, and 2/3 of the pitch of the double helix. As a result, large cavities are formed in three out of six channels surrounding each DNA molecule where the large grooves are juxtaposed. Protamine molecules are also proposed to have some secondary/tertiary structure prior to complex formation. Inside the channels, a protamine molecule modifies its shape to fill the large grooves of all of the three surrounding DNA molecules simultaneously, and might possibly be in touch with other protamine molecules in neighbouring positions as well. This disposition allows the protamine molecules to be located between DNA molecules without a significant increase in the lattice parameters. PMID- 10376016 TI - The hip fracture threat. PMID- 10376017 TI - Pharmacists' outreach visits and doctors' prescribing. PMID- 10376018 TI - Bacteraemia in young children with high fever: still no easy answers. PMID- 10376019 TI - Improving organ donor rates. PMID- 10376020 TI - "Don't confuse me with facts...": evidence-based practice confronts reality. PMID- 10376021 TI - Health burden of hip and other fractures in Australia beyond 2000. Projections based on the Geelong Osteoporosis Study. AB - OBJECTIVE: To calculate the expected increase in the number of fractures in adults attributable to the predicted increase in the number of elderly Australians. DATA SOURCES: All fractures in adult residents (> or = 35 years) of the Barwon Statistical Division (total population, 218,000) were identified from radiological reports from February 1994 to February 1996. The Australian Bureau of Statistics supplied predictions of Australia's population (1996 to 2051). MAIN OUTCOME MEASURE: The projected annual number of fractures in Australian adults up to 2051 (based on stable rates of fracture in each age group). RESULTS: The number of fractures per year is projected to increase 25% from 1996 to 2006 (from 83,000 fractures to 104,000). Hip fractures are projected to increase 36% (from 15,000 to 21,000) because of a substantial rise in the number of elderly aged 85 years and over. Hip fractures are expected to double by 2026 and increase fourfold by 2051. CONCLUSIONS: In contrast to Europe and North America, where numbers of hip fractures are expected to double by 2026 and then stabilise, in Australia hip fractures will continue to place a growing demand on healthcare resources for many decades. These projections can be used for setting goals and evaluating the costs and benefits of interventions in Australia. PMID- 10376022 TI - Outcomes of an educational-outreach service for community medical practitioners: non-steroidal anti-inflammatory drugs. AB - OBJECTIVE: Exploration of longer-term outcomes of an ongoing educational-outreach service for community doctors. DESIGN: Quasi-experimental, with parallel and historical comparisons. SETTING: Since 1992, a teaching-hospital-based service has been providing advice and information on drugs and therapeutic strategies to community medical practitioners. PARTICIPANTS: 210 doctors practising in a particular area of metropolitan Adelaide (79% general practitioners; 21% specialists). INTERVENTIONS: Two surgery visits during 1992 focused on better use of prescribed non-steroidal anti-inflammatory drugs (NSAIDs). Subsequent visits on other topical therapeutic issues have occurred regularly. MAIN OUTCOME MEASURES: Doctor participation in the service; supply of prescription NSAIDs; hospital admissions for gastrointestinal (GI) effects of NSAID use. RESULTS: 89% of doctors practising within the service area received the first visit on NSAIDs and 86% received the second visit. More than 85% continue to receive the service. Relative to a comparison area, aggregate reductions of 9% and 28%, respectively, were observed in two different measures of NSAID use. During an 11-year observation period, a single change point in the number of hospital admissions for GI disorders occurred in the service area, coinciding with delivery of the NSAID program. In the five years since the visits commenced, a 70% reduction in admissions was observed. No notable changes in hospital admission rates occurred in the comparison area. CONCLUSIONS: A continuing education and support service for community medical practitioners which uses principally academic detailing methods in its contact with doctors has contributed to sustained changes in prescribed NSAID use over a five-year period. A focus on risk-minimisation in prescribing of NSAIDs appears to have contributed to reductions in hospitalisations for GI adverse events. PMID- 10376023 TI - Bacteraemia in febrile children presenting to a pae3iatric emergency department. AB - OBJECTIVE: To determine the prevalence of bacteraemia in young febrile children presenting to a paediatric emergency department. DESIGN: Prospective observational case study. SETTING: Emergency Department of the Royal Children's Hospital, Melbourne, between May 1996 and May 1997. PARTICIPANTS: Patients aged 3 36 months presenting to the Emergency Department with temperature > or = 39 degrees C and without specific viral illnesses (varicella, croup or herpes gingivostomatitis). OUTCOME MEASURES: Bacteraemia (defined as presence of pathogenic bacteria in a blood culture), white blood cell count (WCC), McCarthy score, and final diagnosis based on clinical features and investigations. RESULTS: Bacteraemia was identified in 18 of 534 patients (3.4%). Pathogens isolated were Streptococcus pneumoniae (15), Neisseria meningitidis (2) and Klebsiella pneumoniae (1). Increased WCC counts (P < 0.001) and brief duration of fever (P < 0.001) were associated with bacteraemia. Nevertheless, clinical features, including McCarthy scores, and high WCC counts (> or = 20 x 10(9)/L) had < 10% predictive accuracy for bacteraemia. Overall, final diagnoses in the 534 febrile children included non-specific viral infections (25%), upper respiratory tract infections (24%), otitis media (10%), gastroenteritis (9%), pneumonia (7%), and urinary tract infection (5%). CONCLUSIONS: Most urban Australian children aged 3-36 months presenting to a paediatric emergency department with temperature > or = 39 degrees C without a clinical focus have a viral infection. However, 3%-4% have occult bacteraemia. Neither clinical features nor high WCC counts reliably identify these patients. As empiric antibiotics may contribute to increasing antibiotic resistance and have not been shown to prevent the rare complication of meningitis, we believe that close contact and regular review of these patients is preferable to empiric antibiotic therapy. PMID- 10376024 TI - Organ donor index: a benchmark for comparing hospital organ donor rates. AB - OBJECTIVE: To develop organ donor indices to assess donor rates of individual hospitals. DESIGN: Data from hospital databases were retrospectively reviewed for patient separation ICD-9-CM codes (i.e., diagnostic codes from the International classification of diseases, 9th revision, clinical modification) to identify and categories actual and potential organ donors. Organ donor indices for groups of codes and for individual hospitals were determined by dividing the number of actual donors by the total number of patients who died with the same separation ICD-9-CM codes. SETTING: The three South Australian adult tertiary hospitals in 1988-1995. PATIENTS: The 154 actual organ donors, and all patients aged less than 71 years who died with the same groups of ICD-9-CM codes as the organ donors. RESULTS: Organ donors could be classified by three groups of ICD-9-CM codes specifying diseases or pathological processes that could result in brain death. These groups were head injury (44.2% of donors), cerebrovascular accident (CVA) (42.2%), and eight "other" codes (13.6%). Differences between the head injury donor indices for the three hospitals were not significant (Hospital A, 19.1%; Hospital B, 24%; Hospital C, 21%), but there were significant interhospital differences in donor indices for the CVA group (A, 11.2%; B, 5.7%; C, 5.1%; P < 0.05) and the "other" group (A, 3.6%; B, 0.7%; C, 0.3%; P < 0.001). CONCLUSIONS: ICD-9-CM codes can be used to describe organ donors and hospital populations from which potential organ donors may be found. The casemix-controlled organ donor indices can be used to compare the organ donor rates of individual hospitals and to examine reasons for low rates (other than purely casemix variation). PMID- 10376025 TI - Disseminated tuberculosis: still a diagnostic challenge. AB - Disseminated tuberculosis is notoriously difficult to diagnose and, with the decrease in tuberculosis incidence in Australia, familiarity with its manifestations has dwindled. We describe four bacteriologically proven cases which illustrate the range of presentations and diagnostic difficulties. Surprisingly, immunosuppressive therapy need not cause rapid deterioration. Disseminated tuberculosis should be considered in any patient with multisystem illness who is at risk of tuberculosis, particularly if born overseas. In the absence of confirmatory results, a prompt therapeutic trial may be life-saving. PMID- 10376026 TI - The potential effect on hip fracture incidence of mass screening for osteoporosis. AB - With ageing of the Australian population, treatment of osteoporosis-related hip fractures will impose an increasing burden on the healthcare system. Based on current age-adjusted hip fracture incidence and population projections for New South Wales, we estimated a 90% increase in hip fractures by the year 2021. Contributing significantly to this increase will be the number of men reaching the high risk age group for osteoporotic hip fractures. A suggested solution- screening and appropriate therapy for individuals at high risk of osteoporosis- may have only a modest impact. Our calculations show that, even with optimistic screening and therapy compliance rates, hip fractures could still increase by over 50%. Other approaches need to be further explored. PMID- 10376028 TI - Reproductive aspects of cancer treatment: an update. PMID- 10376027 TI - How best to fix a broken hip. Fractured Neck of Femur Health Outcomes Project Team. AB - OBJECTIVES: To develop evidence-based guidelines for the treatment of proximal femoral fractures to optimise functional outcome while minimising length of stay in hospital. DATA SOURCES: Systematic literature search of MEDLINE and CINAHL computer databases, bibliographies, and current contents of key journals for 1966 1995. STUDY SELECTION: English-language randomised controlled trials of all aspects of acute-care hospital treatment of proximal femoral fracture among subjects aged 50 years and over with proximal femoral fractures not due to metastatic disease. DATA EXTRACTION: Two independent reviewers, blinded to authors, institution and study results, followed a standard Cochrane Collaboration protocol and assessed study quality and treatment conclusions. When necessary, a third review was performed to reach consensus. RESULTS: Of the 120 articles published between 1966 and December 1995, 97 met the inclusion criteria. Fifteen clinical interventions were reviewed. Five were supported by National Health and Medical Research Council (NHMRC) level I evidence (prophylactic anticoagulants, prophylactic antibiotics, regional anaesthesia, pressure relieving mattresses, and internal surgical fixation), two had no supporting randomised controlled trial evidence (time to surgery, time to mobilisation after surgery) and the remainder were classified as having Level II evidence. A review of current practice (1993-94) identified wide variability in these interventions across five acute-care hospitals in the Northern Sydney Area Health Service. CONCLUSIONS: Randomised controlled trial evidence (NHMRC Levels I and II) exists for many, but not all, aspects of hip fracture treatment. There is a need for changes to be made to some aspects of practice in accordance with evidence-based guidelines. PMID- 10376029 TI - Cardiology. 4. Atrial fibrillation. PMID- 10376030 TI - GPs and clinical audit. PMID- 10376031 TI - RACOG and CME. Royal Australian College of Obstetricians and Gynaecologists. PMID- 10376032 TI - Performance and outcome measures. PMID- 10376033 TI - Australia's anaesthetists are world leaders. PMID- 10376034 TI - SA's Perioperative Mortality Committee. PMID- 10376035 TI - Commonwealth initiatives. PMID- 10376036 TI - Acute copper poisoning from drinking lime cordial prepared and left overnight in an old urn. PMID- 10376037 TI - Advances in gastrointestinal endoscopy. PMID- 10376038 TI - Indiscriminate use of aciclovir. PMID- 10376039 TI - Factors associated with waiting time for surgery. PMID- 10376040 TI - Doctors detected self-administering opioids in New South Wales, 1985-1994: characteristics and outcomes. PMID- 10376041 TI - Irukandji envenomation in far north Queensland. PMID- 10376042 TI - Should medical students read Plato? PMID- 10376043 TI - Are we headed for a surplus of 'generalist' healthcare providers? It might be time to pull the plug on incentives. PMID- 10376044 TI - Lyme disease vaccine safe for all? PMID- 10376045 TI - Why different forms of testosterone? PMID- 10376046 TI - Is Y2K really such a big problem? Tips on preparing your clinical practice. PMID- 10376047 TI - Marijuana and cancer risk. PMID- 10376048 TI - What's normal for glucose? PMID- 10376049 TI - Work-related asthma and latex allergy. Sorting out the types, causes, and consequences. AB - Work-related asthma now has clear definitions based on criteria agreed upon by the American College of Chest Physicians. The clinician should suspect occupational asthma, irritant-induced asthma, or work-aggravated asthma in adults with new-onset asthma or asthma symptoms that worsen during work, after work (late allergic response), or over the course of workdays. The possible cause should be sought, and a skin test or immunoassay should be performed, if possible,to he;lp detect sensation. Workup als o includes objective documentation of worsening of symptoms and airway obstruction during occupational exposure. If this information is inconclusive, an inhalation challenge may be considered. Medical management is the same as for nonoccupational asthma, but cessation of exposure to the specific agent is necessary to improve long-term diagnosis. Latex allergy and latex-induced asthma are becoming more common in the workplace, particularly in the healthcare field. No commercially available standard serum or skin test are available for diagnosis. The principal treatment is avoidance of latex, which can be achieved in most cases without extensive changes to the workplace. PMID- 10376050 TI - Flexible sigmoidoscopy. Illuminating the pearls for passage. AB - As efforts to improve screening for colon cancer have increased, so has the importance of routine flexible sigmoidoscopy. However, the procedure is still underused, and about half of primary care physicians who recommend it refer their patients to other physicians. Increased use of this valuable screening tool is an important goal in primary care practice. Drs Davis and Stanfield discuss the indications, contraindications, and goals of the procedure as well as special techniques and maneuvers that ensure its safety and efficiency. PMID- 10376051 TI - Practical management of pleural effusions. When and how should fluid accumulations be drained? AB - Management of pleural effusions requires recognition of the cause and initiation of disease-specific therapy. However, it is important to be forewarned that breathing problems do not always improve in these patients, since other disease processes may be contributing to dyspnea. Many options are available for draining effusions and preventing recurrence, but decisions often need to be made quickly so the process does not progress to the point at which intervention becomes much more difficult. PMID- 10376052 TI - The painful, protruding eye. Unilateral euthyroid Graves' ophthalmopathy. PMID- 10376053 TI - Practical pointers for grappling with GERD. Heartburn gnaws at quality of life for many patients. AB - GERD is a common condition, generally caused by transient LES relaxations. The spectrum of disease due to GERD is wide, ranging from symptoms alone to esophagitis, Barrett's esophagus, and respiratory tract complications. Excellent diagnostic test are available to confirm the diagnosis and stage the disease, and medications can predictably relieve symptoms and heal even the most severe forms of esophagitis. Finally, surgical therapy is an effective option for patients with truly refractory disease or for those patients with significant disease who prefer not to use drug treatment. PMID- 10376054 TI - Esophageal cancer and Barrett's esophagus. How to approach surveillance, treatment, and palliation. AB - Esophageal cancer is an increasingly common problem with poor survival rates in patients who present with symptoms. The underlying cause for the progressive rise in the incidence of this cancer remains to be determined. Reducing mortality to requires either early identification of patients or prevention for progression from Barrett's esophagus to cancer. Significant questions remain regarding the cost effectiveness of endoscopic and nonendoscopic methods of surveillance. For local esophageal cancer, the traditional approach has been surgical resection. Radiation therapy is sometimes used alone, but chemotherapy alone is not helpful. Combination therapy consisting of chemotherapy along with surgery or radiation may be the best choice. A new option being tried in disease limited to the mucosa is ablation of neoplastic tissue with endoscopic techniques. Treatment of advanced-stage esophageal cancer is limited and may be hampered by the presence of micrometastatic disease. Morbidity and quality-of-life issues need to be considered and discussed with patients, given the current short survival time of most patients with esophageal cancer. PMID- 10376055 TI - When it's hard to swallow. What to look for in patients with dysphagia. AB - Swallowing disorders can be divided into oropharyngeal dysphagia and esophageal dysphagia. The most common cause of oropharyngeal dysphagia is cerebrovascular accidents; other causes may include oropharyngeal structural lesions, systematic and local muscular diseases, and diverse neurologic disorders. Esophageal dysphagia may result from neuromuscular disorders, mortality abnormalities, and intrinsic or extrinsic obstructive lesions. Through clinical history taking helps define the tpe of dysphagia and can guide diagnostic testing. Important questions to ask patients with the disorder include specific features of the dysphagia, its onset and progression, accompanying problems, and eating habits adopted to relieve symptoms. Videofluoroscopy should be the initial test in evaluating oropharyngeal dysphagia. Barium-contrast esophagography identifies most anatomic causes of dysphagia and some motor disorders and is better tha endoscopy at identifying extrinsic esophageal compression and intramural lesions not involving the esophageal mucosa. Cine-esophagography may provide clues to a possible esophageal motor disorder causing dysphagia. Endoscopy is the test of choice if obstruction or gastroesophageal reflux disease is suspected, because biopsies can confirm the presence of esophagitis and provide specific pathologic identification of the obstructive lesion. In addition, therapeutic dilatation of a stricture and removal of foreign bodies can be accomplished as part of the evaluation procedure. When no obvious source of dysphagia is apparent after radiologic and endoscopic assessment, manometry for possible motility disorder should be considered. PMID- 10376056 TI - The dangers of atypical mole (dysplastic nevus) syndrome. Teaching at-risk patients to protect themselves from melanoma. AB - Everyone with atypical mole syndrome is at increased risk for malignant melanoma. Care of these patients should focus on lowering this risk and on early detection and treatment if melanoma develops. Prophylactic excision of all atypical nevi is not recommended. Patients should be warned of the potential hazards of sun exposure, educated in methods of protecting themselves and their children from the sun, and encouraged to do monthly skin self-examinations. Total-skin examinations at regular intervals should be performed by a physician in patients with atypical mole syndrome. Frequency depends on the specific risk factors, but a total-skin examination should be completed at least once each year, beginning around puberty, and should be continued for life. PMID- 10376058 TI - Adult degenerative joint disease of the knee. Maximizing function and promoting joint health. Institute for Clinical Systems Integration. PMID- 10376057 TI - Using acellular pertussis vaccines for childhood immunization. Potential benefits far outweigh potential risks. AB - If acellular vaccines can offer protection against pertussis that is similar to that afforded by whole-cell vaccines and do so with reduced adverse effects, why have these agents not completely replaced the whole-cell preparations? This article provides an overview of general characteristics of both types of vaccine and some concerns and issues that remain in many physicians' minds. In addition, the authors furnish a rationale and recommendations for safe and appropriate use of the newer acellular formulations. PMID- 10376059 TI - 'Resistant' nail fungus. Are you fighting the real invader? PMID- 10376060 TI - Heartburn and GERD. PMID- 10376061 TI - The making of a curriculum. PMID- 10376062 TI - Feature selection for computerized mass detection in digitized mammograms by using a genetic algorithm. AB - RATIONALE AND OBJECTIVES: To investigate optimization of feature selection for computerized mass detection in digitized mammograms, and to compare the effectiveness of a genetic algorithm (GA) in such optimization with that of an "exhaustive" search of all feature permutations. MATERIALS AND METHODS: A Bayesian belief network (BBN) was used to classify positive and negative regions for masses depicted in digitized mammograms; 20 features were computed for each of 592 positive and 3,790 negative regions in two databases. Conditional probabilities for the BBN were computed by using a "training" database of 288 positive and 2,204 negative regions. Performance was measured by the area under the receiver operating characteristic curve (A) by using the remainder database (304 positive and 1,586 negative regions). The optimal set was first found by using an "exhaustive" (complete permutation) searching method. A GA-based search for the optimal set then was applied, and the results of the two approaches were compared. RESULTS: As the number of features in the classifier increased, the A value increased until it reached a maximum performance for 11 features of 0.876 +/- 0.008. The A value then decreased monotonically as the number of features increased from 11 to 20. Using 100 random chromosomes (seeds) in the first generation, the GA identified the same optimal set of features but reduced the total computation time by a factor of 65. CONCLUSION: A GA-based search might be an efficient and effective approach to selecting an optimal feature set. PMID- 10376063 TI - Touch-preparation cytologic examination of breast core biopsy specimens: accuracy in predicting benign or malignant core histologic results. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to determine the accuracy of touch-preparation cytologic examination of breast core biopsy specimens in predicting benign or malignant core histologic results. MATERIALS AND METHODS: One hundred two core biopsies were performed on 88 women with stereotactic or ultrasonographic (US) guidance. Slides were prepared by smearing one core sample on each slide, spraying the slides with fixative, and staining them with the Papanicolaou technique. Slides were blindly reviewed by a cytopathologist. Cytologic results were categorized as positive for malignancy, not diagnostic for malignancy, or insufficient for diagnosis. Results were correlated with histologic results from all specimens obtained during the core biopsy. RESULTS: Imaging depicted the lesions sampled for biopsy as masses (n = 70), clustered calcifications (n = 29), focal asymmetries (n = 2), or architectural distortion (n = 1). Touch-preparation slides of 87 (85%) lesions contained sufficient material for diagnosis. Cytologic results correctly identified 12 of 16 (three of five intraductal and nine of 11 invasive) malignancies in 10 of 13 masses and two of three clusters of calcifications. Two false-positive results occurred, both with fibroadenomas. Overall, touch-preparation studies produced 69 true-negative and four false-negative results. Excluding slides with insufficient material, the sensitivity, specificity, and accuracy of touch-preparation results were 75%, 97%, and 93%, respectively. Including insufficient samples, accuracy was 79%. CONCLUSION: Although touch-preparation cytologic examination of breast core biopsy specimens is fairly accurate in prediction of benign or malignant core histologic results, its correlation with histologic results is not sufficient to justify routine use in immediate counseling and treatment planning. PMID- 10376064 TI - Volume (three-dimensional) fast spin-echo imaging of the lumbar spine. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to prospectively evaluate a proton-density-weighted, three-dimensional (3D) volume fast spin-echo (SE) pulse sequence in the assessment of the lumbar spine for suspected spondylosis. MATERIALS AND METHODS: Twenty-eight patients referred for low back or lower extremity pain were imaged with both a two-dimensional (2D) protocol and a proton density-weighted 3D volume fast SE imaging. The spinal canal, conus medullaris, intervertebral disks, neural foramina, bone marrow, and spinal alignment shown with the 3D volume fast SE pulse sequence were independently assessed by two neuroradiologists. These findings were compared with those of the routine 2D studies. RESULTS: Interpretation of disk protrusions and stenoses of the neural foramina were concordant between both protocols. No instance of cord abnormality was detected with either protocol. CONCLUSION: A 3D volume fast SE proton-density weighted pulse sequence may provide information comparable to that of routine 2D imaging. Advantages of volume imaging include thinner sections, the capability of reconstruction into any plane, and the potential to decrease imaging time. PMID- 10376065 TI - In vivo evaluation of the adjustable temporary venous spring filter and the RF02 temporary filter: comparative study. AB - RATIONALE AND OBJECTIVES: The authors performed this study to compare the in vivo efficacies of the temporary venous spring filter and the RF02 filter in an animal model. MATERIALS AND METHODS: Either the spring filter or the RF02 filter was placed in the inferior vena cava of 10 pigs each, and two clots (5 x 20 mm) were funneled into the filters at 1-hour intervals. The second clots were funneled without removing the first clots captured by the filters. Clot-trapping ability, caval occlusion associated with the clot-trapping procedure, arterial blood gas concentrations, and changes in arterial and iliac venous pressures were evaluated. RESULTS: Placement of the RF02 filter caused elevation of iliac venous pressure with a maximum of 2.2 mm Hg (median) (n = 13, P = .003). Placement of the spring filter parallel to venous flow enabled capture of 90% (nine of 10) and 100% (six of six) of the first and second clots, respectively. The RF02 filter captured clots consistently. The difference between filters was not statistically significant. Both filters equally contributed to elevation of iliac venous pressure (median, 9.3 and 7.2 mm Hg [n = 9] with the spring filter and RF02 filter, respectively). Caval occlusion occurred in 17% (one of six) and 67% (six of nine) of animals after two clots were trapped in the spring filter and RF02 filter, respectively (P = .06). Other parameters were not influenced by the clot trapping procedure. CONCLUSION: Although a larger version should be developed and better stability of the filter is needed, the spring filter proved to be an efficient filtering device and had a lower rate of caval occlusion compared with the RF02 filter. PMID- 10376066 TI - Automated calculation of the centerline of the human colon on CT images. AB - RATIONALE AND OBJECTIVES: This article presents an evaluation of an automated technique for determining the colon centerline with computed tomographic (CT) data sets. MATERIALS AND METHODS: The technique proceeds as follows. After indication of a voxel in the rectum, voxels corresponding to air were segmented. Points along the colon centerline were estimated on the basis of centers of mass of grown voxels. A second segmentation and centerline calculation was initiated at the cecum. These two centerlines were then averaged. The resulting average was refined by using lumen data obtained perpendicular to the average centerline. The accuracy of the technique was investigated with simulation phantoms. The technique was also evaluated for 40 clinical colon cases. Calculated centerline points were compared with those indicated by radiologists for a randomly selected clinical case. RESULTS: In the simulation studies, the calculated centerline points were, on average, within 2.5 mm of the true centerlines but differed by up to 4 mm in regions of deep folds or sharp turns. In the clinical colon study, 40% of the centerlines were computed with a single seed point and 25% with two seed points. Average centerlines were computed in 1 minute. The root mean square difference between the computed centerline points and those indicated by the radiologists was 4-5 mm (comparable to interobserver variations). CONCLUSION: Accurate centerlines can be determined from colon CT data with this automated technique. PMID- 10376068 TI - Research in interventional MR imaging: where do we go from here? PMID- 10376067 TI - Musculoskeletal radiology curriculum guide. The Education Committee of the American Society of Musculoskeletal Radiology. PMID- 10376069 TI - Do's and don't's of percutaneous nephrostomy. AB - Percutaneous nephrostomy procedures generally are safe. The associated mortality rate is approximately 0.04%, and the incidence of important complications is 5% (2-4). To minimize complications, certain precautions always should be followed. First, radiologists should perform a preprocedural evaluation of the patient, with correction of marked coagulopathy or thrombocytopenia before all but the most emergent procedures. Second, antibiotics should be administered routinely before nephrostomy drainage; the choice of antibiotics can be based on the specific patient's risk factors for bacteriuria. To minimize the risk of clinically important renal vascular damage, radiologists should do the following: 1. Always achieve adequate visualization of the calices. 2. Identify a posterior calix for puncture that will give access to the appropriate segment of the kidney for anticipated procedures and allow safe creation of a tract. 3. Puncture below the 11th rib (and preferably below the 12th rib when feasible). 4. Puncture the tip of a posterior calix from a 20 degrees-30 degrees, posterolateral oblique approach to avoid major blood vessels. 5. Make a single-wall puncture of the calix. 6. Perform exchange transfusion for opacification of the renal pelvis and calices during percutaneous nephrostomy procedures to minimize the risk of sepsis. Overdistention can increase the likelihood of sepsis or retroperitoneal contamination. 7. Inject contrast material via a catheter placed over a wire to confirm the intracollecting system location of the entry. 8. Avoid unnecessary (complicated, prolonged) procedures in an infected, obstructed system. 9. Use only self-retaining drainage catheters to minimize the risk of inadvertent catheter dislodgment. 10. Create large-bore tracts with a balloon dilation system. By contrast, radiologists should not do the following: 1. Puncture above the 11th rib (unless all other avenues of approach have been exhausted). 2. Lose access to an obstructed kidney once the kidney has been punctured. Placement of a "safety" wire for all complex manipulations is recommended. 3. Panic if excessive bleeding or evidence of adjacent organ injury is seen. Excessive bleeding usually can be stopped with tract tamponade by using a balloon catheter advanced through the tract or with placement of an appropriate-sized nephrostomy tube to occlude the tract. If active bleeding continues or recurs, arteriography should be considered. The quantity of bleeding can be monitored with sequential hematocrit measurements. Almost all renal artery injuries can be treated with minimally invasive procedures, such as selective embolization of the branch artery involved, and this will lead to infarction of only a small segment of kidney, with preservation of functioning renal parenchyma. Injury to an adjacent organ usually can be treated nonsurgically (21,23). The most commonly injured extrarenal abdominal organ is the colon (Fig 6). On occasion, a percutaneous nephrostomy needle may traverse the retroperitoneal segment of the colon, and this type of injury generally can be treated nonsurgically, as well (23). If the colon has been traversed, adequate urinary drainage should be ensured before the transcolonic nephrostomy catheter is removed (so that a nephrocolonic fistula is not maintained). This can be done by placing a ureteral stent and a bladder catheter (18). Once adequate urinary drainage is provided, the nephrostomy catheter can be withdrawn into the colon and used as a percutaneous colostomy drain. The percutaneous colostomy tract should be allowed to mature for several days before this catheter is removed. In addition, appropriate antibiotics should be administered from the time a transcolonic tract is identified until the percutaneous tract has healed completely. Transthoracic entry can cause pneumothorax and pleural effusions. These should be treated only if they are large or cause symptoms (21). (ABSTRACT TRUNCATED) PMID- 10376070 TI - Report of the Working Group on Digital Mammography: Computer-Aided Diagnosis and 3D Image Analysis and Display. Cambridge, Massachusetts, USA. October 8-9, 1998. PMID- 10376071 TI - Heavy metal poisoning and its laboratory investigation. PMID- 10376072 TI - Measurement of selenium in clinical specimens. PMID- 10376073 TI - Assessing the quality of comments on reports: a retrospective study. AB - I describe a comparatively simple method for assessing the quality of comments on clinical biochemistry reports. It is based on independent peer review of components of comments, appears reasonably robust and correlates well with separate independent assessment of the value of complete comments. The method gives a numerical score, and thus lends itself to a wide range of statistical manipulations and assessment for education and audit purposes. PMID- 10376074 TI - Carotenoid composition and antioxidant potential in subfractions of human low density lipoprotein. AB - Carotenoids and vitamin E are transported in human plasma complexed with lipoproteins. The bulk of them are associated with low-density lipoprotein (LDL), in which form they may act as antioxidants and thus delay the onset of atherosclerosis. We used a simple, rapid, ultracentrifugation technique to fractionate plasma lipoproteins in self-generating gradients of iodixanol (Optiprep), a non-ionic iodinated density gradient medium. The carotenoid content and composition of a number of LDL subfractions was determined by reversed-phase high-performance liquid chromatography. Lycopene, beta-carotene and beta cryptoxanthin were mainly located in the larger, less-dense LDL particles whereas lutein and zeaxanthin were found preferentially in the smaller, more dense LDL particles. When the antioxidant content of these fractions was expressed per milligram of LDL protein, significantly lower concentrations of carotenoid and vitamin E were found to be associated with the smaller, protein-rich fractions of LDL. Strong positive correlations were found between total carotenoid and vitamin E plasma concentrations and the lag-time of Cu(2+)-mediated oxidation of LDL subfractions. The more dense LDL subfractions, which had lower levels of these antioxidants, were more readily oxidized, highlighting their possible role in atherosclerotic events. PMID- 10376075 TI - Effect of low-density lipoprotein (LDL) antigen source on an enzyme-linked immunosorbent assay for autoantibodies against oxidized LDL. AB - We examined the effect of antigen source on an enzyme-linked immunosorbent assay (ELISA) for autoantibodies against oxidized low-density lipoprotein (LDL). Serum samples from 20 subjects with systemic lupus erythematosus (SLE) and from 20 controls were assayed for immunoglobulin G (IgG) and immunoglobulin M (IgM) autoantibodies against oxidized LDL, using either a pooled or individual (n = 3) LDL preparation as antigen. For IgG autoantibodies against oxidized LDL there was a relationship (r approximately 0.5, P < 0.01) between data obtained using individual versus pooled antigen preparations. Bias plots demonstrated consistent inverse, concentration-dependent relationships (r approximately -0.6, P < 0.001). The difference in IgG autoantibodies against oxidized LDL levels between SLE patients and controls was underestimated (39-58%) when assays used individual rather than pooled LDL antigen. For IgM autoantibodies against oxidized LDL the direct relationships were stronger (r approximately 0.8, P < 0.001) and the concentration-dependent relationships weaker (r approximately -0.3, significance variable) than for IgG autoantibodies against oxidized LDL. Variations between LDL preparations suggested that a pooled antigen would give a more stable assay. Thus, LDL antigen source is important in assays for both IgG and IgM autoantibodies against oxidized LDL. PMID- 10376076 TI - Influence of investigative and operative procedures on serum prostate-specific antigen concentration. AB - We determined the effect of cystoscopy (flexible and rigid), transrectal ultrasonography (with and without needle biopsy of the prostate) and transurethral resection of the prostate or bladder tumour on the serum prostate specific antigen (PSA) concentration. Samples were taken from 60 men before and up to 14 days following these procedures. Flexible cystoscopy did not result in a significant increase in serum PSA concentration, with a median increase of 0.1 microgram/L (P > 0.05). Small but statistically significant increases in serum PSA levels 1 day post-procedure were observed following rigid cystoscopy and transrectal ultrasound without biopsy. The median increase in serum PSA concentration following rigid cystoscopy was 0.15 microgram/L (P = 0.04) and following transrectal ultrasound was 0.3 microgram/L (P = 0.01). In both cases the serum PSA level had normalized by 2 days post-procedure. Transurethral resection of bladder tumours resulted in a variable rise in serum PSA, with a median increase of 2.6 micrograms/L after 1 day, which returned to normal over 7 14 days. Ultrasound-guided needle biopsy of the prostate and transurethral resection of the prostate produced significant increases in serum PSA levels, which took up to fourteen days to return to normal. The median increase in serum PSA following needle biopsy was 6.0 micrograms/L and following transurethral resection of the prostate (TURP) was 13 micrograms/L. Samples for PSA measurement may safely be taken within 24-48 h of flexible cystoscopy and transrectal ultrasonography (TRUS) providing prostatic biopsy is not carried out. For other procedures it is necessary to wait for at least 14 days to ensure that false positive PSA results are not obtained. PMID- 10376077 TI - Threshold concentration of unbound bilirubin to induce neurological deficits in a patient with type I Crigler-Najjar syndrome. AB - Based on the clinical course of a 16-year-old boy with type I Crigler-Najjar syndrome, we estimated the threshold concentration of unbound bilirubin, as assayed by the horseradish peroxidase method, that apparently induces toxicity to the brain. Before the age of 15, the patient did not manifest any neurological or behavioural dysfunction despite increased bilirubin in serum. The binding affinity and the binding capacity of the patient's serum albumin for bilirubin determined when he was about 14 years old were 10(8)(mol/L)-1 and 1.01 to 1.04 mol/L, respectively. These values were nearly the same as those of normal controls reported in the literature. The total bilirubin binding capacity was greater than the patient's total bilirubin concentration, showing that his serum albumin was not saturated with bilirubin. The reserve bilirubin binding capacity (RBBC) was estimated to be 158 mumol/L and the unbound bilirubin concentration to be 15.1 nmol/L. Concentration of unbound bilirubin peaked at 21.7 nmol/L at the age of 15 years and 11 months, i.e. 2 months before the onset of difficulties in walking and speaking. At this time, the RBBC was estimated as -64 mumol/L. A peak concentration of total bilirubin, 811 mumol/L, was observed during the period of rapid loss of the ability to walk or speak. At the age of 16 years and 1 month the RBBC decreased to -98 mumol/L and the unbound bilirubin concentration to 18.8 nmol/L. Following phototherapy, the patient's neurological state returned to normal; he could speak and walk normally. At the age of 16 years and 2 months the RBBC returned to 105 mumol/L and unbound bilirubin decreased to 16.6 nmol/L. These results suggest that maintaining the concentration of unbound bilirubin at < 20 nmol/L and the total bilirubin concentration at lower than the binding capacity of serum albumin is important for prevention of neurological deficits in Crigler-Najjar syndrome. The upper limit of unbound bilirubin in such an older patient was nearly the same as that reported for newborns. PMID- 10376078 TI - Eosinophil cationic protein in serum from nonatopic and asymptomatic atopic individuals after standardized blood clotting at 37 degrees C. AB - Eosinophil cationic protein (ECP) and ECP/eosinophil ratio were measured in 223 apparently healthy subjects. The serum sample for ECP measurement was obtained by standardized clotting of the blood sample for 2 hours at 37 degrees C. Eosinophil count, ECP concentration and ECP/eosinophil ratio were no different between men (n = 122) and women (n = 101). Reference ranges for serum ECP and ECP/eosinophil ratio were 12-99 micrograms/L and 61-367 micrograms ECP per 10(9) eosinophils, respectively. Serum ECP and blood eosinophil count were positively correlated (y = 141x + 18, R2 = 0.45, P < 0.001). The ECP/eosinophil ratio, however, was found to decrease with increasing eosinophil count. Twenty-three per cent of the apparently healthy subjects were found to have a positive score for immunoglobulin E antibodies specific to inhalant allergens, and thus were considered atopic. Serum ECP concentrations and eosinophil counts were significantly higher in this group compared with the non-atopic group. In this healthy population no decision level for either eosinophil count, serum ECP or ECP/eosinophil ratio could be found that discriminated atopic from non-atopic individuals. PMID- 10376079 TI - Evaluation of different weight correction methods for antenatal serum screening using data from two multi-centre programmes. AB - Weight correction of serum markers is widely used when screening for Down's syndrome and open neural tube defects (NTD) because marker concentrations decrease with increasing maternal weight. Log-linear regression is frequently used for determining weight correction factors, but recently reciprocal-linear regression has been suggested to have advantages. We compared both methods of weight correction using data from two screening programmes carried out by this laboratory, one using alpha-fetoprotein (AFP) and total human chorionic gonadotrophin (HCG) (n = 129,143) and the other, AFP and free beta-HCG (n = 39,982). The reciprocal-linear method fitted the data more closely but did not significantly alter the detection rate or screen positive rate (SPR) for Down's syndrome or NTD with either dataset. Without correction, women heavier or lighter than average weight had significantly different SPRs for Down's syndrome and NTD compared with those weighing close to the median weight. Both correction methods smoothed out the variability in the SPR for Down's syndrome to a similar degree, but reciprocal-linear regression was much better at reducing the variability in SPR for NTD and its use is therefore worthwhile. PMID- 10376080 TI - Detecting anti-SSA and anti-SSB antibodies in routine analysis: a comparison between double immunodiffusion and immunoblotting. AB - The aim of this study was to assess the performance of a commercially available procedure for detecting anti-Sjogren's syndrome A (anti-SSA) and anti-Sjogren's syndrome B (anti-SSB) antibodies by immunoblotting (IB) and compare it with double immunodiffusion (DID). We also studied the clinical significance of these profiles in a series of unselected anti-SSA positive patients. Serum samples from 534 patients that were positive on an immunofluorescent screening test using HEp 2 cells were analysed for anti-SSA and anti-SSB antibodies by DID and IB (Biolab Anablot System II), and the results on anti-SSA antibodies were confirmed by an enzyme-linked immunosorbent assay (ELISA). Fifty-five serum samples were found to be positive for anti-SSA antibodies. Among these, 24 were anti-SSA negative by IB but positive by DID and ELISA ('non-blotter sera'), whereas only three serum samples were anti-SSA negative by DID but positive by IB and ELISA. Of the 18 anti-SSB positive serum samples, eight were negative by DID. All the serum samples that were anti-SSB positive by DID were also positive by IB. Anti-SSB antibodies showed a significant association with eye dryness and leucopenia. Anti 52 kDa SSA antibodies were associated with anti-SSB antibodies but showed no significant association with sicca symptoms, while anti-60 kDa SSA antibodies were associated with lower rates of leucopenia. The 'non-blotter' profile showed no significant association with any clinical parameter. IB is less sensitive than DID for detecting anti-SSA antibodies but more sensitive than DID for detecting anti-SSB antibodies. The determination of anti-SSA immunoblotting profiles in patients positive for anti-SSA antibodies by DID does not significantly improve the clinical usefulness of this test. As expected, anti-SSB antibodies were associated with clinical features of Sjogren's disease. Non-blotting (probably conformational) anti-SSA antibodies did not show any further association with clinical parameters and seem to have no clinical relevance. PMID- 10376081 TI - Recommended approaches for the laboratory measurement of homocysteine in the diagnosis and monitoring of patients with hyperhomocysteinaemia. AB - Several recent studies have indicated that an increased concentration of plasma homocysteine is an independent risk factor for the premature development of vascular disease. These important findings emphasize the need for careful selection of an appropriate analytical approach to diagnose and treat individuals who may be at risk. We compared the results obtained from the measurement of plasma total homocysteine (free + protein-bound fractions) by high-performance liquid chromatography (HPLC) with the measurement of plasma free homocystine (free fraction) by conventional ion-exchange chromatography in 10 patients with inherited defects of homocysteine metabolism and 13 obligate heterozygote individuals. This study can be used to formulate recommendations on the appropriate use of these assays in different clinical circumstances. Our results show that the concentration of total plasma homocysteine must exceed 60 mumol/L before plasma free homocystine becomes detectable by conventional ion-exchange chromatography. Similarly, assessment of the urinary excretion of homocysteine in these patients indicates that it may not become consistently detectable by conventional ion-exchange chromatography or HPLC until plasma total homocysteine exceeds 150 mumol/L. On this basis, while most patients with classical homocystinuria would be detected by analysis of plasma using conventional ion exchange chromatography or by measurement of of the urinary homocysteine excretion, occasional patients would be missed. When monitoring patients receiving treatment for classical homocystinuria, in whom metabolic control is good, and when investigating individuals with a suspected inherited defect of cobalamin or folate metabolism, a method which measures plasma total homocysteine should be used. The identification of moderate hyperhomocysteinaemia of undefined cause investigated in relation to a history of early vacsular disease can only be identified by this approach. PMID- 10376082 TI - Evaluation of six erythropoietin kits. AB - Six erythropoietin (EPO) measurement kits from different manufacturers were evaluated for ease of use and performance. Reference values were compared from patient samples with normal and abnormal circulating serum EPO. The kits evaluated were EPO-Trac RIA, EPORIA, DSL RIA, EPO EIA, IBL ELISA and Medac ELISA (see below for manufacturers' details). The radioimmunoassay (RIA) methods were simple to perform with little preliminary preparation of reagents but the non isotopic methods were of varying degrees of complexity. Imprecision data showed that performance of the RIA methods was acceptable across the range 40-150 mU/mL [within-assay coefficient of variation (CV) = 2-5%, between-assay CV = 3-7%] but poor (CV > 10%) at the lower end of the range (10-15 mU/mL). The IBL enzyme linked immunosorbent assay (ELISA) kit showed good precision across the range 10 54 mU/mL (within-assay CV = 5-7%, between-assay CV = 8-12%). The performance of the EPO enzyme immunoassay (EIA) and Medac ELISA were acceptable although precision was poor at the lower end of the range (< 10 mU/mL; within-assay CV 10% and 17% and between-assay CV 13% and 18%, respectively). The ELISA and EIA methods had lower limits of detection (0.4-0.8 mU/mL) than the RIA methods (2.3 3.6 mU/mL). Analysis of serum EPO measurement variation between methods showed evidence of significant negative bias with DSL RIA and Medac ELISA when compared with EPO-Trac RIA. Conversely, EPORIA showed significant positive bias and the two remaining methods, EPO EIA and IBL ELISA, showed no evidence of systematic bias when compared with EPO-Trac RIA. Patients with normal circulating concentration of serum EPO gave values that were within the reference range of the kits. PMID- 10376083 TI - Rapid detection of insulin-like growth factor-binding protein-1 and foetal fibronectin in cervico-vaginal secretions to diagnose premature membrane rupture. PMID- 10376085 TI - Is the Bayer DCA 2000 acceptable as a screening instrument for the early detection of renal disease? PMID- 10376084 TI - Association of the 'secretor state' with the presence and recurrence of urinary infections in pregnant women. PMID- 10376086 TI - Night blindness following hemicolectomy and radiotherapy. PMID- 10376087 TI - Let's talk about purchasing power. PMID- 10376088 TI - The international world of nursing. PMID- 10376090 TI - Caring for the Islamic patient. AB - The delivery of culturally sensitive care by perioperative nurses is an essential element of patient advocacy. To provide culturally astute care, nurses must familiarize themselves with the world's religious and ethnic groups. Islam is a worldwide religion and, like all religions, is practiced along a spectrum ranging from very conservative and traditional practices to the more liberal and contemporary ones. A person may accept some, all, or none, of the principles discussed in this manuscript. Additionally, the patient's country of origin plays an integral role in the planning of culturally competent care. PMID- 10376089 TI - Airborne particulates in the OR environment. AB - Intraoperative sampling of airborne particulates is rarely performed in the OR environment because of technical difficulties associated with sampling methodologies and because of the common belief that airborne contamination is infrequently associated with surgical site infections (SSIs). In this study, investigators recovered non-viable (i.e., lint) and viable (i.e., microorganisms) particulates during vascular surgery using a personal cascade impactor sampling device. The predominant nonviable particulates recovered during intraoperative sampling were wood pulp fibers from disposable gowns and drapes. Several potential nosocomial pathogens (e.g., Staphylococcus aureus, Staphylococcus epidermidis) and other drug-resistant isolates frequently were recovered from an area adjacent to the surgical field. The widespread presence of airborne particulates during surgery suggests that further studies are warranted to assess the role these particles may play in the development of SSIs or in dissemination of nosocomial pathogens within the OR and hospital environment. PMID- 10376092 TI - An introduction to nursing research through an OR nursing journal club. PMID- 10376091 TI - Perioperative management of burn patients. AB - Burns continue to be a leading cause of death in the United States. Survivability of burn patients has increased since 1988 because of significant changes in the management of life-threatening injuries. Advances include rapid removal of eschar, skin grafting, and early enteral feeding. Pathophysiologic changes occur in every major organ system of the burn patient and must be aggressively treated. Perioperative care of the burn patient begins in the immediate postburn period and continues throughout the patient's care. Intraoperative management of the patient requires organization, planning, and, above all, communication between the OR team members and anesthesia care providers to ensure optimum results. PMID- 10376093 TI - Who can be called a "nurse"? PMID- 10376094 TI - Internet resources for perioperative care of burns. PMID- 10376095 TI - Using ethical analysis when there is no research. PMID- 10376096 TI - Comparison of intradermal testing and serum testing for allergen-specific IgE using monoclonal IgE antibodies in 84 atopic dogs. AB - OBJECTIVE: To compare an ELISA measuring serum allergen-specific IgE with intradermal skin testing in canine atopic dermatitis. PROCEDURE: Eighty-four dogs with the clinical diagnosis of atopic dermatitis underwent intradermal skin testing and serum testing for allergen-specific IgE. Tests were performed in a blinded fashion. Positive reactions were compared and the sensitivity and specificity of the serum test (using intradermal skin test as the standard) were determined overall and for individual allergen groups (grass pollens, weed pollens, tree pollens, house dust mites and fleas). RESULTS: The sensitivity of the ELISA overall was 90.4%. Evaluating the individual allergen groups, the sensitivity for dust mite hypersensitivity was 95.1%, for fleas 85.4%, for tree pollens 84.3%, for grass pollens 95.1% and for weed pollens 96.4%. The specificity was 91.6% overall, for dust mites 96.3%, for fleas 92.7%, for tree pollens 95.2%, for grass pollens 94% and for weed pollens 80.7%. CONCLUSION: The evaluated ELISA seemed reliable for the diagnosis of atopy in practice and can be recommended as a screening test prior to intradermal skin testing or for use in dogs when immunotherapy is not a therapeutic option. PMID- 10376097 TI - Disc extrusion in a Rottweiler dog with caudal cervical spondylomyelopathy after failure of intervertebral distraction/stabilisation. AB - A Rottweiler dog was presented with an 8 week history of hindlimb ataxia. Neurological examination localised the lesion to the cervical spinal cord. Myelography demonstrated dynamic compressive lesions at C5-6 and C6-7 consistent with a diagnosis of caudal cervical spondylomyelopathy. Distraction/stabilisation of both discs was performed using interbody polymethyl methacrylate. Both implants subsequently failed leading to extrusion of the remaining dorsal annulus fibrosus of the C5-6 intervertebral disc and nonambulatory tetraparesis. A ventral slot combined with distraction/stabilisation using screws and polymethyl methacrylate was performed and resulted in nearly full neurological recovery. PMID- 10376099 TI - Total ear canal ablation and lateral bulla osteotomy in an alpaca. PMID- 10376098 TI - Effect of acute haemorrhage on QRS amplitude of the lead II canine electrocardiogram. AB - OBJECTIVE: To examine the effect of acute haemorrhage on the QRS amplitude of the canine lead II surface electrocardiograph (ECG). DESIGN: Ten adult racing Greyhounds were tranquilised, anaesthetised, positioned in right lateral recumbency and connected to recording electrodes of an ECG unit. Baseline six lead ECG traces were recorded, and further traces were obtained after one unit (460 mL) of blood, and then a second unit, were collected from the femoral artery. RESULTS: There was a consistent and progressive reduction in amplitude of the QRS complex in all leads during acute haemorrhage. QRS amplitude in lead II after removal of two units of blood averaged 74% of the baseline voltage, with individual values of 61 to 91% (P < 0.0001). There were even greater reductions in QRS amplitudes in lead aVL during haemorrhage. In three additional dogs, reductions in QRS voltages were shown to be accompanied by reductions in end diastolic left ventricular internal dimensions measured echocardiographically. Furthermore, the effects of haemorrhage on the QRS amplitude and echocardiographic measurements were reversed when circulating blood volume was restored by re-infusion of blood removed previously. CONCLUSION: Acute haemorrhage corresponding to an approximately one-third reduction in blood volume caused a substantial reduction in QRS voltage of the surface ECG. It is postulated that this resulted from diminished ventricular distension as a consequence of reduced venous return. A similar mechanism may account for the small-amplitude ECG complexes associated with pericardial effusion, severe dehydration and hypovolaemia. PMID- 10376100 TI - Outcomes after extrahepatic portosystemic shunt ligation in 49 dogs. AB - OBJECTIVE: To evaluate outcomes after attenuation of extrahepatic portosystemic shunts in dogs using surgical silk. DESIGN: Retrospective study. PROCEDURE: Case records were reviewed for degree of surgical attenuation, experience of the primary surgeon, perioperative mortality and problems related to persistent portosystemic shunting or shunt ligation. Presence of portosystemic shunting after surgery was evaluated by ammonia tolerance testing, measurement of postprandial serum bile acid, plasma urea and cholesterol concentrations and liver enzyme activity. The influence of age, postocclusion portal pressure, primary surgeon, degree of attenuation and postoperative biochemical findings on the occurrence of postoperative problems was assessed. RESULTS: The mortality rate was 2.1%. Shunt attenuation was complete in 34% and partial in 66% of dogs. Portal hypertension necessitating ligature removal was encountered in only one dog. Five dogs experienced neurological abnormalities (seizures or ataxia), possibly as a manifestation of 'postligation seizure syndrome'. Postoperative liver function was normal in 78% of dogs, including 70% with partial shunt attenuation. Experience of the surgeon was related positively to outcome after partial attenuation (P = 0.002). Postoperative biochemical evidence of abnormal liver function was the most sensitive predictor of recurrence of clinical signs referable to persistent portosystemic shunting. CONCLUSIONS: In the hands of an experienced surgeon, surgical attenuation of single extrahepatic shunts was safe and effective, even in animals with partial attenuation. Most dogs with biochemical evidence of persistent shunting suffer relapse of clinical signs within 18 months of surgery. Postligation neurological syndromes of variable intensity may be more common than previously thought. PMID- 10376101 TI - Equine piroplasmosis: the temporary importation of seropositive horses into Australia. PMID- 10376102 TI - Vitamin D doses for alpacas (Lama pacos). AB - OBJECTIVE: To assess the effectiveness of cholecalciferol (D3) doses for maintaining adequate vitamin D status in crias and adult female alpacas at pasture. DESIGN: A field experiment during winter and early spring in a herd on a farm in South Australia. ANIMALS AND PROCEDURE: Crias, usually less than 6 months of age and female alpacas, aged 2 to 6 years, were given a single subcutaneous dose of 0, 1000 or 2000 IU D3/kg body weight. Plasma concentrations of 25 hydroxycholecalciferol (25-OH D3), phosphorus, calcium and vitamins A and E and alkaline phosphatase activity were measured at intervals over a period of 16 weeks after treatment. RESULTS: Crias not given a vitamin D supplement had reduced growth rate during winter and one animal showed clinical signs of rickets. Vitamin D treatment had no effect on the body weight of mature females. Vitamin D supplements increased the 25-OH D3 and phosphorus concentrations in plasma of both crias and adult females; alkaline phosphatase activity was not affected by treatment. CONCLUSION: It is suggested that for alpacas in southern Australia a subcutaneous dose of 1000 IU D3/kg body weight to crias in late autumn and again in mid winter and to adult females in mid winter should prevent vitamin D inadequacy. PMID- 10376103 TI - Plasma cortisol concentrations in normal dogs given hydrocortisone sodium succinate. AB - OBJECTIVE: To evaluate the effects on plasma cortisol concentration of a continuous infusion of a readily available steroid with equipotent glucocorticoid and mineralocorticoid effects. PROCEDURE: Plasma cortisol concentrations were measured before and regularly after hydrocortisone sodium succinate was administered as a continuous intravenous infusion over 6 h at 0.32 and 0.65 mg kg 1 h-1 to 12 healthy dogs weighing 12 to 22 kg. RESULTS: The infusion at both does rates produced significant and stable increases in plasma cortisol concentrations. The plateau concentrations produced by the large and small doeses were respectively above and below plasma cortisol concentrations likely to provide adequate glucocorticoid and mineralocorticoid activity in stressed dogs with significantly decreased adrenal function. CONCLUSION: This paper presents information regarding the changes in plasma cortisol concentrations in 12 normal dogs given an hydrocortisone sodium succinate infusion at two dose rates. The marked and continuous increase in plasma cortisol concentrations suggests a continuous HSS infusion may be a possible alternative to desoxycorticosterone acetate and dexamethasone in the treatment of acute adrenal dysfunction. PMID- 10376104 TI - Effectiveness of small workshops for improving farmers' knowledge about ovine footrot. AB - OBJECTIVE: To determine sheep farmers' attitudes to and beliefs about ovine footrot, and to improve their knowledge about the diagnosis, control and eradication of this disease. METHOD: Eighteen workshops, involving 291 farmers, were conducted across Victoria in the spring of 1996. The workshops were designed as small-group discussions with a maximum attendance of 20 farmers to encourage active participation. All participants completed questionnaires before each workshop and 12 to 40 weeks after the last workshop. RESULTS: Before the workshops the farmers had a poor understanding of the principles of diagnosis, control and eradication of footrot. For example, only 50% knew the footrot organism survived in soil for less than 7 days, over two-thirds did not know the reason for paring sheep's feet during an eradication program, and only 31% realised cattle were a potential source of footrot infection for sheep. After the workshops, understanding about footrot was significantly improved; 87% said Dichelobacter nodosus survived in soil less than 7 days, 71% knew the reason for paring sheep's feet and 64% realised that cattle were a potential source of footrot infection. As well as improved knowledge, change of attitude among farmers is fundamentally important if virulent footrot is to be successfully controlled and eradicated. The workshops successfully initiated this process; 40% of farmers thought the workshops changed their attitudes to footrot, while 37% said they gained an increased understanding of other people's opinions about the disease. CONCLUSION: Farmers' poor understanding of ovine footrot is a constraint to the programs aimed at controlling this disease. Small group workshops may be an effective way to influence farmers' attitudes and beliefs, and could facilitate the effectiveness of regulatory disease control programs. PMID- 10376105 TI - A cross-sectional study of risk factors affecting the outcome of rabbit haemorrhagic disease virus releases in New South Wales. AB - OBJECTIVE: To determine what factors governed the extent of outbreaks of rabbit haemorrhagic disease (RHD) following releases in New South Wales. DESIGN: Retrospective cross-sectional study. PROCEDURE: Information from the data set of official releases was subjected to two preliminary analyses. More comprehensive information on a subsample of official RHD releases, sites and animals was gathered by telephone survey of Rural Lands Protection Board staff and farmers. Data were analysed using multivariate techniques to determine which factors were associated with rabbit mortality within one month of RHDV release, within several months of release and in affecting the proportion of the population killed. RESULTS: A strong association was found between the presence of heavy flea infestation (odds ratio 2.7), breeding in rabbits and outbreaks of RHD. For each week following breeding there was an 8% decline in the odds of an outbreak. Low temperatures also promoted outbreaks. Less important effects included the prior presence of RHD at the release site, which reduced the likelihood and severity of outbreaks. The presence of cattle and proximity to the nearest water body were associated with increased severity and likelihood of outbreaks respectively. CONCLUSION: Both breeding of rabbits and associated high flea numbers may act together or independently in promoting outbreaks of RHD. Stresses involved with rabbit reproduction and low environmental temperatures also appear to influence the likelihood of outbreaks. The effects of proximity to cattle and water suggests that both flies and mosquitoes may have a minor role in local transmission. PMID- 10376106 TI - Autofluorescent substance and neurocyte necrosis in thiamine deficiency in cattle. PMID- 10376107 TI - Serological survey for Brucella antibodies in feral pigs from eastern Australia. PMID- 10376108 TI - Australia's vital assistance to Asian nations fighting serious infectious diseases. PMID- 10376109 TI - Times-and vets-have changed. PMID- 10376110 TI - AVA went 'too far'. PMID- 10376111 TI - Trial by tv & vet fees. PMID- 10376112 TI - DHEA: mood, memory, and aging. PMID- 10376113 TI - Dehydroepiandrosterone treatment of midlife dysthymia. AB - BACKGROUND: This study evaluated the efficacy of the adrenal androgen, dehydroepiandrosterone, in the treatment of midlife-onset dysthymia. METHODS: A double-blind, randomized crossover treatment study was performed as follows: 3 weeks on 90 mg dehydroepiandrosterone, 3 weeks on 450 mg dehydroepiandrosterone, and 6 weeks on placebo. Outcome measures consisted of the following. Cross sectional self-ratings included the Beck Depression Inventory, and visual analogue symptom scales. Cross-sectional objective ratings included the Hamilton Depression Rating Scale, the Cornell Dysthymia Scale and a cognitive test battery. Seventeen men and women aged 45 to 63 years with midlife-onset dysthymia participated in this study. Response to dehydroepiandrosterone or placebo was defined as a 50% reduction from baseline in either the Hamilton Depression Rating Scale or the Beck Depression Inventory. RESULTS: In 15 patients who completed the study, a robust effect of dehydroepiandrosterone on mood was observed compared with placebo. Sixty percent of the patients responded to dehydroepiandrosterone at the end of the 6-week treatment period compared with 20% on placebo. A significant response was seen after 3 weeks of treatment on 90 mg per day. The symptoms that improved most significantly were anhedonia, loss of energy, lack of motivation, emotional "numbness," sadness, inability to cope, and worry. Dehydroepiandrosterone showed no specific effects on cognitive function or sleep disturbance, although a type II error could not be ruled out. CONCLUSIONS: This pilot study suggests that dehydroepiandrosterone is an effective treatment for midlife-onset dysthymia. PMID- 10376115 TI - Magnetoencephalography: applications in psychiatry. AB - Magnetoencephalography (MEG) measures the extracranial magnetic fields produced by intraneuronal ionic current flow within appropriately oriented cortical pyramidal cells. Based upon superconducting quantum interference device technology operating at liquid helium temperatures (4 K), MEG offers excellent temporal and spatial resolution for selected sources, and complements information obtained from electroencephalograms and other functional imaging strategies. Current instrumentation permits recording up to several hundred channels simultaneously with head-shaped dewars, although the cost of such systems is high. The fact that magnetic fields fall off with the square of the distance from the source is both a benefit (when separating activity in the two hemispheres) and a limitation (when attempting to record deep sources). The lack of skin contact facilitates using MEG to record direct current and very high frequency (> 600 Hz) brain activity. The clinical utility of MEG includes presurgical mapping of sensory cortical areas and localization of epileptiform abnormalities, and localization of areas of brain hypoperfusion in stroke patients. MEG studies in psychiatric disorders have contributed materially to improved understanding of anomalous brain lateralization in the psychoses, have suggested that P50 abnormalities may reflect altered gamma band activity, and have provided evidence of hemisphere-specific abnormalities of short-term auditory memory function. PMID- 10376114 TI - Anterior cingulate cortex dysfunction in attention-deficit/hyperactivity disorder revealed by fMRI and the Counting Stroop. AB - BACKGROUND: The anterior cingulate cognitive division (ACcd) plays a central role in attentional processing by: 1) modulating stimulus selection (i.e., focusing attention) and/or 2) mediating response selection. We hypothesized that ACcd dysfunction might therefore contribute to producing core features of attention deficit/hyperactivity disorder (ADHD), namely inattention and impulsivity. ADHD subjects have indeed shown performance deficits on the Color Stroop, an attentional/cognitive interference task known to recruit the ACcd. Recently, the Counting Stroop, a Stroop-variant specialized for functional magnetic resonance imaging (fMRI), produced ACcd activation in healthy adults. In the present fMRI study, the Counting Stroop was used to examine the functional integrity of the ACcd in ADHD. METHODS: Sixteen unmedicated adults from two groups (8 with ADHD and 8 matched control subjects) performed the Counting Stroop during fMRI. RESULTS: While both groups showed an interference effect, the ADHD group, in contrast to control subjects, failed to activate the ACcd during the Counting Stroop. Direct comparisons showed ACcd activity was significantly higher in the control group. ADHD subjects did activate a frontostriatal-insular network, indicating ACcd hypoactivity was not caused by globally poor neuronal responsiveness. CONCLUSIONS: The data support a hypothesized dysfunction of the ACcd in ADHD. PMID- 10376116 TI - A pilot study of penicillin prophylaxis for neuropsychiatric exacerbations triggered by streptococcal infections. AB - BACKGROUND: Some children with obsessive-compulsive disorder (OCD) and tic disorders appear to have symptom exacerbations triggered by group A beta hemolytic streptococcal infections in a manner that is similar to rheumatic fever and its neurologic variant, Sydenham's chorea. Because penicillin prophylaxis has proven to be effective in preventing recurrences of rheumatic fever, it was postulated that it might also prevent streptococcal-triggered neuropsychiatric symptom exacerbations in children with Pediatric Autoimmune Neuropsychiatric Disorders Associated with Streptococcal infections (PANDAS). These children are identified by five clinical characteristics: presence of OCD or tic disorder, prepubertal onset, episodic symptom course, neurologic abnormalities (i.e., choreiform movements) and streptococcal-triggered symptom exacerbations. METHODS: Thirty-seven children with PANDAS were enrolled in an 8 month, double-blind, balanced cross-over study. Patients were randomized to receive either 4 months of the active compound (twice daily oral 250 mg penicillin V) followed by 4 months of placebo, or placebo followed by penicillin V. Tic, OCD, and other psychiatric symptoms were monitored monthly. Throat cultures and streptococcal antibody titers were also obtained. RESULTS: There were an equal number of infections in both the active and placebo phases of the study. There was no significant change seen in either the obsessive-compulsive or tic symptom severity between the two phases. CONCLUSIONS: Because of the failure to achieve an acceptable level of streptococcal prophylaxis, no conclusions can be drawn from this study regarding the efficacy of penicillin prophylaxis in preventing tic or OCD symptom exacerbations. Future studies should employ a more effective prophylactic agent, and include a larger sample size. PMID- 10376117 TI - CSF 5-HIAA levels are lower in impulsive as compared to nonimpulsive violent suicide attempters and control subjects. AB - BACKGROUND: We studied CSF 5-HIAA and HVA concentrations in violent suicide attempters and examined their relationship with depression, anxiety, and impulsivity. METHODS: CSF 5-HIAA and HVA concentrations were determined very shortly after hospital admission and compared to those of a matched control population. Clinical evaluation was performed concomitantly; the level impulsivity was evaluated by the Impulsivity Rating Scale (IRS). RESULTS: Twenty three patients and 23 control subjects were included. According to the IRS, 14 patients were classified as impulsive, including all patients suffering from personality disorders, and 9 as nonimpulsive, with a main DSM-IIIR diagnosis of melancholia. CSF 5-HIAA concentrations in the suicide group were significantly lower than in control subjects. This difference was entirely due to the impulsive suicide attempters. There was an inverse correlation between the IRS score and CSF 5-HIAA (r = -.47, p = .02) and only a trend for HVA (r = -.41, p = .078) levels in the suicide group. CONCLUSIONS: This study of a group of violent suicide attempters distinguished a subgroup of patients diagnosed with personality disorder with high impulsivity scores and a subgroup of patients with the main diagnosis of severe depression. CSF 5-HIAA was significantly lower in impulsive violent attempters than in nonimpulsive violent attempters, therefore desintangling violence from impulsivity and linking this biologic abnormality to impulsivity. PMID- 10376118 TI - Pupillary cholinergic sensitivity to pilocarpine increases in manic lithium responders. AB - BACKGROUND: The cholinergic hypothesis of affective disorders predicts that mania is a hypocholinergic state relative to monoaminergic activity. Treatments that increase cholinergic sensitivity are expected to improve manic symptoms. METHODS: Ten male hypomanic or manic patients were treated with lithium carbonate (0.7-1.1 mEq/L) for 2 weeks. Cholinergic sensitivity was assessed prior to, and following treatments, using graded concentrations of pilocarpine eyedrops (.03-2.0%). Pupil size changes were quantified using an infrared pupillometer and ED50 values were referenced to maximal dilation with .5% tropicamide. RESULTS: Lithium treatment decreased Bech mania ratings and ED50 values (p < .001). Improvements in mania with lithium treatment were closely correlated with decreases in ED50 (r = .88, p < .001). CONCLUSIONS: These results support the cholinergic-adrenergic hypothesis and suggest that one possible mechanism for the antimanic effects of lithium may involve increasing cholinergic activity in relation to monoaminergic neurotransmission. PMID- 10376120 TI - CAG repeat length in the hKCa3 gene and symptom dimensions in schizophrenia. AB - BACKGROUND: Long CAG repeats in the hKCa3 potassium channel gene have been associated with schizophrenia. We sought evidence for associations between this polymorphism and aspects of the schizophrenia phenotype. METHODS: Associations were investigated between CAG repeat length and gender, age of illness onset, and psychotic symptom dimensions in 203 unrelated individuals with DSM-IIIR schizophrenia. RESULTS: No association was found between CAG repeat length and gender or age of onset. Long CAG repeats were associated with higher negative symptom dimension scores. CONCLUSIONS: This study provides preliminary evidence that genetic liability to negative symptoms in schizophrenia may be partly mediated through the hKCa3 gene. PMID- 10376119 TI - Functional PAX-6 gene-linked polymorphic region: potential association with paranoid schizophrenia. AB - BACKGROUND: Early differentiation of the nervous system and adult CNS neuroplasticity is modulated by PAX-6. We have shown previously that a highly polymorphic, functional AC/AG repeat in the 5' regulatory region of the gene showed significantly increased promoter activity, if containing > or = 29 repeats, and that the heterozygous genotype (< or = 28/> or = 29) revealed increased mRNA PAX-6 levels in human brain tissue compared to the homozygous short variant. METHODS: In a case-control study of 655 unrelated individuals, allele frequencies and genotype distributions of the functional PAX-6 promoter polymorphism were investigated comprising patients with DSM-IV schizophrenia, patients with affective disorders, and population controls. RESULTS: No allelic or genotypic association of the PAX-6 promoter polymorphism to affective disorder or to schizophrenia as one disease entity was observed. After subtyping schizophrenia into paranoid and nonparanoid forms, potential evidence was found for a genotypic association of the high-activity variant with the paranoid subtype of schizophrenia (p = .02). The estimated odds ratio was 1.7 (95% CI .98 to 2.95) for those heterozygous and 1.4 (95% CI .82 to 2.42) for those heterozygous or homozygous for the high-activity variant compared to the homozygous low-activity variant. CONCLUSIONS: Our finding indicates that early developmental genes may be involved in the etiopathogenesis of schizophrenia subtypes via variable transcriptional regulation in the developing and adult human brain. PMID- 10376121 TI - Volumetric analysis of frontal lobe regions in schizophrenia: relation to cognitive function and symptomatology. AB - BACKGROUND: The purpose of this study was to examine the structure of dorsolateral, medial, and orbital regions of the frontal lobe in schizophrenia, and to determine whether their volumetric measurements were related to cognitive function and symptomatology. METHODS: High resolution magnetic resonance imaging scans of the brains of 14 schizophrenic patients and 14 closely matched healthy controls were acquired. Volumes of gray and white matter of the left and right dorsolateral, medial, and orbital prefrontal brain regions were measured. Tests of verbal and visual memory and executive functions were used to assess cognitive function. The SANS and SAPS were used to obtain symptom ratings in patients. RESULTS: Data of 13 schizophrenic patients were analyzed. Patients showed a general, though not significant, decrease in volumes of frontal regions as compared to controls. In patients, but not in controls, smaller left and right prefrontal gray matter volumes were significantly correlated with impaired performance on immediate recall in verbal and visual memory and semantic fluency. Furthermore, in patients, smaller total orbitofrontal gray matter volume was significantly correlated with more severe negative symptomatology (rs = -.76, p = .006). CONCLUSIONS: These findings suggest that in schizophrenia, deficits in verbal and visual memory and semantic fluency and negative symptoms may be related to (subtle) abnormalities in frontal lobe structure. PMID- 10376122 TI - Prolactin hyperresponsiveness to D-fenfluramine in drug-free schizophrenic patients: a placebo-controlled study. AB - BACKGROUND: Functional alterations in the central serotonergic system have been reported in schizophrenia but no conclusive data have been provided. In the present study, we investigated the prolactin (PRL) response to the selective serotonin (5-HT) releasing agent D-fenfluramine in both patients with schizophrenia and matched healthy subjects. METHODS: Sixteen drug-free schizophrenics and 16 healthy subjects were randomized in a double-blind neuroendocrine test to D-fenfluramine (30 mg p.o.) or placebo. Blood PRL and cortisol concentrations were determined by radioimmunoassay, while plasma levels of D-fenfluramine were measured by mass spectrometry. RESULTS: In schizophrenic patients, baseline plasma PRL levels were not different from controls, whereas plasma cortisol concentrations were significantly increased (p < .03). The PRL response to D-fenfluramine was significantly enhanced in patients as compared to matched control subjects (p < .005). Schizophrenics meeting Kane's criteria for previous nonresponse to typical neuroleptics exhibited a PRL response to D fenfluramine significantly higher than non-drug-resistant patients (p < .04). No significant difference in plasma D-fenfluramine concentrations was observed between schizophrenic and healthy subjects. CONCLUSIONS: These findings suggest a serotonergic hypersensitivity in chronic schizophrenia. This alteration seems to be peculiar to those patients refractory to typical neuroleptics. PMID- 10376123 TI - Topography of the auditory P300 in schizotypal personality. AB - BACKGROUND: Research with schizophrenic patients has demonstrated reduced amplitude of the P300b elicited with the auditory "oddball" paradigm, as well as reduced P300a amplitude following "novel" stimuli. The focus of the present study was the investigation of these components in a nonclinical sample of participants with high expressions of the schizotypal personality trait. METHODS: By use of an acoustic oddball task, including the presentation of novel stimuli, the event related brain potentials of 14 participants with "low" and 13 participants with "high" scores on the German adaptation of the "Schizotypal Personality Questionnaire" were investigated. Current source density (CSD) curves and spline interpolated CSD maps were generated. Peak amplitudes and latencies of the N100, P200, P300a, and P300b were determined for the CSD data. RESULTS: Results indicate no group differences with respect to N100, P200, and P300a amplitudes and latencies. By contrast, the P300b amplitude was significantly smaller in high as compared to low-scoring participants. Left-temporal as compared to right temporal P300b was significantly smaller in high- than in low-schizotypal participants. CONCLUSIONS: Confirming results of other researchers, this present study suggests that a reduced P300b amplitude and an altered P300b topography at temporal sites may be a trait-like "marker" of the schizophrenia spectrum. PMID- 10376124 TI - Effects of continuous exposure to light on behavioral dopaminergic supersensitivity. AB - BACKGROUND: This study examines the effects of long-term continuous exposure to light on dopaminergic supersensitivity induced by repeated treatment with haloperidol in rats. METHODS: Spontaneous general activity in an open-field (SGA) and stereotyped behavior induced by apomorphine (SB-APO) or amphetamine (SB-AMP) were used as experimental parameters. Rats were allocated to four groups in each experiment: saline-treated animals kept under a 12-hour light/dark cycle (LD) or 24-hour light/light cycle (LL), and 2 mg/kg haloperidol-treated animals kept under the above cycles. Plasma corticosterone concentration was also measured by radioimmunoassay in saline-treated rats kept under a LD or LL cycle. RESULTS: All the behavioral parameters used showed the development of central dopaminergic supersensitivity in rats kept under both cycles. Continuous exposure to light enhanced SGA and SB-AMP in both saline- and haloperidol-treated rats, but did not modify SB-APO. Animals kept under the LL cycle presented an increased plasma corticosterone concentration. CONCLUSIONS: Our results suggest that continuous exposure to light leads to an increase in dopaminergic function in both normal and "supersensitive" rats. This effect seems to be mediated by a presynaptic mechanism possibly involving corticosterone actions. PMID- 10376125 TI - Choreoathetoid movements in cocaine dependence. AB - BACKGROUND: To evaluate the severity of choreoathetoid movements in cocaine dependent (CD) subjects and age-matched normal control subjects. METHODS: Choreoathetoid movements were evaluated using the Abnormal Involuntary Movement Scale (AIMS) in samples of 71 CD, 56 normal control, and 9 amphetamine-dependent male subjects. RESULTS: The CD subjects had a significantly increased nonfacial (limbs plus body) AIMS subscore. When the nonfacial AIMS scores of the two groups were compared in relation to age, a significant age by diagnosis interaction was observed, indicating that the differences between groups were most marked in the younger age groups. The facial AIMS scores were also increased but only in the youngest CD cohort (under 32 years of age). The comparison group of 9 younger amphetamine-dependent subjects also showed increased AIMS scores. CONCLUSIONS: Increases in choreoathetoid movements in younger cocaine and amphetamine dependent subjects may be related to their psychostimulant use. The absence of differences in choreoathetoid movements between the older CD subjects and normal control subjects may represent an age-related self-selection effect. PMID- 10376126 TI - The effects of naltrexone maintenance on the response to yohimbine in healthy volunteers. AB - BACKGROUND: Preclinical research suggests that opiate antagonists may alter stress responsiveness. This study describes the effect of pretreatment with the opioid antagonist naltrexone on the response to a noradrenergic stressor, the alpha-2-receptor-antagonist, yohimbine, in healthy subjects. The current study was designed to compare the change in responses to yohimbine after 2 weeks of treatment with naltrexone to the response after at least 2 weeks of treatment with placebo. METHODS: After a week of placebo naltrexone treatment, ten subjects were randomized into a double-blind cross-over to placebo or active naltrexone (50 mg p.o. daily) on weeks 2 to 4, and the converse condition for weeks 5 to 7. Subjects received challenges in a random, fixed sequence with placebo and active yohimbine (i.v., 0.2 mg/kg) on weeks 1, 4, and 7. The active-active combination generally had the strongest drug effects. RESULTS: There were statistically significant (p < .05) interactions of naltrexone condition X yohimbine condition for subject ratings of "nervous," "not liking the drug effect," "talkative," and "urge to urinate," and a trend (p < .10) for cortisol levels. CONCLUSIONS: The results suggest that clinically used naltrexone doses alter sensitivity to yohimbine. PMID- 10376127 TI - Daily rhythm of serum melatonin levels and effect of light exposure in patients with dementia of the Alzheimer's type. AB - BACKGROUND: Several studies have suggested that patients with dementia experience a deterioration of biological rhythms. We investigated the daily profile of serum melatonin levels in patients with dementia of the Alzheimer's type (AD), since daily melatonin rhythm is thought to reflect the functioning of the biological clock. METHODS: Seventeen inpatients with AD, 10 psychiatric inpatients without dementia, and 11 elderly healthy volunteers participated in this study. Serum melatonin was assessed every 3 hours by radioimmunoassay. RESULTS: A daily fluctuation of melatonin levels with a significant nocturnal increase was observed in all three subject groups. However, both the AD patients and psychiatric patients without dementia showed significantly higher levels of melatonin in the daytime compared with the healthy subjects. When the effect of bright light exposure on melatonin secretion was investigated in six AD patients and five psychiatric patients without dementia, the daytime levels were markedly decreased in the patients without dementia, while no change was observed in the AD patients. CONCLUSIONS: The high levels of melatonin in the daytime associated with a lack of response to light exposure in AD patients may be due to the neurodegenerative process of this disease. PMID- 10376128 TI - Risperidone-induced increase of plasma norepinephrine is not correlated with symptom improvement in chronic schizophrenia. AB - BACKGROUND: Previous studies have shown an increase in plasma levels of norepinephrine (NE) after clozapine treatment of schizophrenia. This effect has been suggested to relate to improvement in symptoms. METHODS: To test whether other novel antipsychotic drugs have such an effect, the present experiment examined schizophrenic symptoms and plasma levels of NE before and after 5 weeks of treatment with risperidone or haloperidol. RESULTS: Risperidone, but not haloperidol, significantly increased plasma NE; however, there was no correlation of this effect with clinical improvement on any symptom scale. CONCLUSIONS: This finding suggests that risperidone shares similar properties with clozapine in enhancing peripheral NE, but that these changes in plasma NE may not be a consistent indicator of atypical antipsychotic drug efficacy. PMID- 10376129 TI - EEG correlates of methylphenidate response among children with ADHD: a preliminary report. AB - BACKGROUND: Recent electrophysiologic studies have found fairly consistent differences between children with attention-deficit/hyperactivity disorder (ADHD) and age-matched control subjects. The present study examined electroencephalogram (EEG) changes associated with a double blind, placebo-controlled administration of methylphenidate among children with ADHD. METHODS: Subjects were 10 children, ages 8 to 13, with a primary diagnosis of ADHD. Brain electrical activity was recorded with 7 electrodes in the frontal, central, and midline areas during baseline and cognitive activation conditions. RESULTS: Repeated-measures ANOVAs indicate that children exhibiting a positive medication response had reductions of theta and alpha as well as increased beta in the frontal regions, while nonresponders showed the opposite pattern (p < .05). Significant correlations between improvement on a vigilance task and changes in beta activity in the frontal electrodes emerged as well. CONCLUSIONS: These preliminary findings indicate that there are different electrophysiologic correlates to methylphenidate among ADHD children who are medication responders and nonresponders. PMID- 10376130 TI - Differential display combined with a regulated transient expression system. PMID- 10376131 TI - Effect of growth temperature and vector ends phosphorylation in the cloning efficiency of specific DNA fragments. PMID- 10376132 TI - Instability of a multiple copy enhancer in plasmid vectors: practical considerations. PMID- 10376133 TI - Functional testing of a bicistronic retroviral vector for intracellular peptide production. PMID- 10376134 TI - Vectors for inducible expression of dual epitope-tagged proteins in insect cells. PMID- 10376135 TI - Screening procedure for Pichia pastoris clones containing multiple copy gene inserts. PMID- 10376136 TI - Direct cloning of differential display products eluted from northern blots. PMID- 10376137 TI - Simple protocol for extracting nuclear DNA from single embryos of a marine snail. PMID- 10376138 TI - Extracting high-quality DNA from shed reptile skins: a simplified method. PMID- 10376139 TI - Recycling of anion-exchange resins for plasmid DNA purification. PMID- 10376140 TI - Removal of ethidium bromide and cesium chloride from plasmid DNA by ethanol precipitation. PMID- 10376141 TI - Color coding the cell death status of plant suspension cells. PMID- 10376142 TI - Morphological detection of plasma membrane changes during apoptosis using enhanced green fluorescent protein. PMID- 10376143 TI - Extending the useful life span of DNA probes for fluorescence in situ hybridization. PMID- 10376144 TI - Microtiter immunocytochemical ELISA assay. PMID- 10376145 TI - Improved protocol for using avian red blood cells as substrates for the polymerase chain reaction. PMID- 10376146 TI - Bridge-overlap-extension PCR method for constructing chimeric genes. PMID- 10376147 TI - Microsatellite locus amplification using deer antler DNA. PMID- 10376148 TI - Snapping up SNPs. PMID- 10376150 TI - Rapid genetic screening for hemochromatosis using automated SSCP-based capillary electrophoresis (SSCP-CE). AB - Hereditary hemochromatosis (HHC) represents an autosomal recessive disease in which increased iron absorption causes iron overload and irreversible tissue damage. The recently detected association between two point mutations in the HFE gene on chromosome 6p and HHC has made it possible to screen for the disease before the onset of irreversible tissue damage. Conventional genetic testing is based on restriction fragment-length polymorphisms (RFLP) using two endonuclease recognition sites in codon 63 or 282, respectively. In this study, we have adapted single-strand conformation polymorphism analysis for capillary electrophoresis (SSCP-CE) to detect homozygote or heterozygote point mutations. Two HFE gene fragments spanning codons 63 and 282 were amplified by a duplex PCR using genomic DNA from peripheral blood or from tissue sections of paraffin embedded liver biopsies as template. Thereby, rapid genotyping of both HFE mutations was achieved with a single PCR, omitting the need of further analysis by restriction digest. Eighty-five patients with liver disease and/or suspected iron overload were genotyped using SSCP-CE, and all results were verified by conventional RFLP analysis. In summary, SSCP-CE proved to be a reliable, cost effective, sensitive and rapid method for genotyping HFE mutations. This method will further facilitate high-throughput genetic screening using capillary array electrophoretic devices. PMID- 10376149 TI - Differential sensitivity to 5-fluoro-orotic acid as a screen for bait RNA independent false positives in a yeast three-hybrid system. AB - The yeast three-hybrid system presents a valuable tool for detecting and analyzing RNA-protein interactions in vivo. A major drawback of the use of such a transcriptional reporter-based assay in a library screen is the frequent occurrence of false-positive results due to bait RNA-independent activation of the reporter gene. To minimize the isolation of false positives in three-hybrid library screens, we incorporated a rapid and simple procedure based on differential sensitivity to 5-fluoro-orotic acid. The technique effectively eliminates bait RNA-independent false positives and thus greatly enhances the efficiency of the yeast three-hybrid system. PMID- 10376151 TI - Absolute quantitation of MDR1 transcripts using heterologous DNA standards- validation of the competitive RT-PCR (CRT-PCR) approach. AB - The multidrug resistance (MDR1) gene product, P-glycoprotein (Pgp), is a 170-kDa ATP-dependent pump that expels a variety of anticancer drugs out of malignant cells, reducing drug accumulation and thus antitumor activity. In recent years, considerable data has been presented that indicates the need to standardize detection methods for Pgp and MDR1. Reverse transcription (RT)-PCR is one of the most sensitive and specific techniques used to detect MDR1. Nevertheless, there is the need to address working criteria for quantitation by RT-PCR. In this study, we describe a flexible assay used to quantify MDR1 gene expression using heterologous (nonhomologous) standards for use in competitive RT-PCR (CRT-PCR). Our guidelines were to use a RT-PCR quantitation method that was independent of exponential phase kinetics, sensitive (detect low levels of gene measurement in clinical samples) and did not require radiolabel. Furthermore, the method would need to be flexible enough for the user to express quantitation as either the number of cells or amount of cDNA used in CRT-PCR. Using low-stringency amplification, heterologous DNA competitors were constructed for MDR1 and as an internal reference, the ubiquitously expressed human histone variant 3.3 (H3.3). The benefits of this approach are threefold: (i) amplification kinetics of target and competitor molecules are identical, (ii) low-stringency PCR is a simple way of constructing heterologous DNA competitors that do not require special storage conditions and (iii) heterologous competitors avoid the formation of heteroduplex molecules. We conclude that CRT-PCR is an extremely flexible and sensitive assay that can quantify MDR1 based on competitive amplification of a heterologous competitor. This might complement future efforts to standardize MDR1 detection methods using RT-PCR. PMID- 10376152 TI - Model system for plant cell biology: GFP imaging in living onion epidermal cells. AB - The ability to visualize organelle localization and dynamics is very useful in studying cellular physiological events. Until recently, this has been accomplished using a variety of staining methods. However, staining can give inaccurate information due to nonspecific staining, diffusion of the stain or through toxic effects. The ability to target green fluorescent protein (GFP) to various organelles allows for specific labeling of organelles in vivo. The disadvantages of GFP thus far have been the time and money involved in developing stable transformants or maintaining cell cultures for transient expression. In this paper, we present a rapid transient expression system using onion epidermal peels. We have localized GFP to various cellular compartments (including the cell wall) to illustrate the utility of this method and to visualize dynamics of these compartments. The onion epidermis has large, living, transparent cells in a monolayer, making them ideal for visualizing GFP. This method is easy and inexpensive, and it allows for testing of new GFP fusion proteins in a living tissue to determine deleterious effects and the ability to express before stable transformants are attempted. PMID- 10376153 TI - Fast and reliable screening of mutations in human tumors: use of multiple fluorescence-based long linker arm nucleotides assay (mf-LLA). AB - Human tumor samples were screened for point mutations by adapting a mobility shift assay to automated DNA sizing. This screen identifies the type of point mutation and relative amount of mutated DNA sequences present in a sample. Test samples having known hypoxanthine-guanine phosphoribosyl transferase (hprt)/exon 3 sequence mutations were characterized by: (i) PCR amplification, (ii) fluorescent dye-primer extension with 36-atom linker derived deoxycytosine or deoxyuridine triphosphate and the remaining three natural nucleotides and (iii) sizing of the resulting fluorescently labeled modified strands, using an automated DNA sequencer. Routinely, a range of sizes is observed among the sequence variants of a single DNA target sequence. This is because nucleotide analogs are incorporated into DNA strands in a sequence-dependent manner, resulting in composition-dependent electrophoretic mobility. Thus, point mutations are identified as shifts in mobility between the fluorescently labeled modified strands of the control and test samples. The twenty different hprt/exon 3 single-base substitution mutations tested were easily identified, even at fourfold dilution with control DNA. PMID- 10376154 TI - Construction of gene targeting vectors from lambda KOS genomic libraries. AB - We describe a highly redundant murine genomic library in a new lambda phage, lambda knockout shuttle (lambda KOS) that facilitates the very rapid construction of replacement-type gene targeting vectors. The library consists of 94 individually amplified subpools, each containing an average of 40,000 independent genomic clones. The subpools are arrayed into a 96-well format that allows a PCR based efficient recovery of independent genomic clones. The lambda KOS vector backbone permits the CRE-mediated conversion into high-copy number pKOS plasmids, wherein the genomic inserts are automatically flanked by negative-selection cassettes. The lambda KOS vector system exploits the yeast homologous recombination machinery to simplify the construction of replacement-type gene targeting vectors independent of restriction sites within the genomic insert. We outline procedures that allow the generation of simple and more sophisticated conditional gene targeting vectors within 3-4 weeks, beginning with the screening of the lambda KOS genomic library. PMID- 10376155 TI - Identification of relative protein bands in polyacrylamide gel electrophoresis (PAGE) using a multi-resolution snake algorithm. AB - In polyacrylamide gel electrophoresis (PAGE) image analysis, it is important to determine the percentage of the protein of interest of a protein mixture. This study presents reliable computer software to determine this percentage. The region of interest containing the protein band is detected using the snake algorithm. The iterative snake algorithm is implemented in a multi-resolutional framework. The snake is initialized on a low-resolution image. Then, the final position of the snake at the low resolution is used as the initial position in the higher-resolution image. Finally, the area of the protein is estimated as the area enclosed by the final position of the snake. PMID- 10376156 TI - WinGene/WinPep: user-friendly software for the analysis of amino acid sequences. AB - WinGene1.0/WinPep1.2 is a pair of Microsoft Windows programs designed to read nucleotide or amino acid sequence data. These versatile programs have the following capabilities: (i) searches for open reading frames and their translation, (ii) assisting the design of primers for PCR and (iii) calculation of molecular weight, isoelectric point and molar absorbtion coefficients of polypeptides. Furthermore, hydropathic plots and helical wheel displays are easily produced. The programs run with an intuitive Windows interface, contain a comprehensive help file and enable data exchange with other applications by means of the Copy&Paste command. The software is free for academic and noncommercial users. PMID- 10376157 TI - Digital image processing for rapid analysis of differentially expressed transcripts on high-density cDNA arrays. AB - Usage of filter arrays is becoming increasingly attractive for many research laboratories involved in determination of gene-expression profiles. However, analysis of numerous spots, representing genes or partial gene sequences (ESTs), is still tedious work involving the ordered analysis of vast amounts of numerical tabular data. We present a rapid and efficient method for the visual identification of differentially expressed targets on high-density cDNA filter arrays using standard laboratory equipment and standard software, which is available for free. The method we introduce provides an inexpensive alternative, and no changes in the experimental set up are required. Our results were verified by densitometric analyses performed with an established system. PMID- 10376158 TI - Spreadsheet-based program for alignment of overlapping DNA sequences. AB - Molecular biology laboratories frequently face the challenge of aligning small overlapping DNA sequences derived from a long DNA segment. Here, we present a short program that can be used to adapt Excel spreadsheets as a tool for aligning DNA sequences, regardless of their orientation. The program runs on any Windows or Macintosh operating system computer with Excel 97 or Excel 98. The program is available for use as an Excel file, which can be downloaded from the BioTechniques Web site. Upon execution, the program opens a specially designed customized workbook and is capable of identifying overlapping regions between two sequence fragments and displaying the sequence alignment. It also performs a number of specialized functions such as recognition of restriction enzyme cutting sites and CpG island mapping without costly specialized software. PMID- 10376159 TI - ScienceLabDatabase: a computer program to organize a molecular biology laboratory. AB - A description of a novel laboratory management software is provided. ScienceLabDatabase (SLD) offers a useful platform to organize a molecular biology research laboratory. The program manages stocks of biological samples, including plasmids, antibodies and cell lines, laboratory protocols and addresses, and it includes an easy ordering and funds managing system. Preformed data sheet templates help to store and maintain valuable information on samples or reagents and to facilitate the transfer of accurate information between researchers. Password protection regulates access, and simple button functions allow the use of this system without prior database knowledge. SLD is based on FileMaker Pro Version 4.0, which allows easy customization and import of preexisting data from other applications. The SLD program was successfully tested in several independent research groups and proved a useful tool to efficiently organize a molecular biology laboratory. PMID- 10376160 TI - Platelet nitric oxide metabolites in migraine. PMID- 10376161 TI - NSAIDs. PMID- 10376163 TI - High-density assessment of the IHS classification criteria for migraine without aura: a prospective study. AB - Fifty-six adult female patients with the clinical diagnosis of MwoA kept a diary 6 times per day for 10 consecutive weeks to record the occurrence, pain characteristics, and accompanying symptoms of headache. In order to avoid bias due to retrospection or expectancy the diary was programmed into palmtop computers which signaled the patients with a beep to enter the diary with a random-fixed time schedule: two signals occurred in, respectively, the morning, the afternoon, and the evening, but at different times for each day. The palmtop computers also warranted flawless data storage and automatic computations of response delay and missing values. Of the 339 attacks, 75% had a duration of 4-72 h and 94% confirmed the International Headache Society classification criteria for MwoA concerning pain characteristics and accompanying symptoms. Our results obtained for attacks in treated patients are highly comparable with the results of Rasmussen, Jensen, and Olesen (1991) obtained in the general population with unknown treatment of headache. Together, both studies support the IHS classification criteria for MwoA. The electronic Experience Sampling Method also allowed for an unbiased description of the course of treated MwoA attacks: 67% subsided in the first day. In the 16 attacks the characteristics and accompanying symptoms were present in 60-80% of the attacks at the first assessment (9.30 a.m.) with the exception of moderate to severe pain intensity (37%) and nausea or vomiting (31%). A waxing and waning of characteristics and symptoms over the day remained in about 30-40% of the attacks with a tendency towards increases in the evening (7.30 p.m. and 10 p.m.). The method is there for a replication of this study in untreated MwoA patients. PMID- 10376164 TI - Familial occurrence of chronic tension-type headache. AB - Chronic tension-type headache (CTTH) assessed by proband report was evaluated in a family study of CTTH. A clinical interview of first-degree relatives by a physician was used as index of validity. Familial occurrence of CTTH in first degree relatives was also investigated. Patterns of familial aggregation of CTTH were assessed by calculating the population relative risk. A neurological resident carried out all the interviews of probands and their first-degree relatives. The operational diagnostic criteria of the International Headache Society were used. The 122 probands had 377 first-degree relatives. Sensitivity, specificity, predictive values, and chance-corrected agreement rate for the diagnosis CTTH were 68%, 86%, 53% (PVpos), 92% (PVneg), and 0.48, respectively. The low sensitivity of CTTH assessed by proband report indicates that a clinical interview by a physician is necessary in family studies of CTTH. Clinically interviewed parents, siblings, and children had a 2.1 to 3.9-fold significantly increased risk of CTTH compared with the general population. The gender of the probands did not influence the risk of CTTH among first-degree relatives. The significantly increased familial risk of CTTH and no increased risk of CTTH in spouses suggest that a genetic factor is involved in CTTH. PMID- 10376165 TI - Comorbidity of headache and depressive disorders. AB - The goal of the present study was to investigate the clinical profile of patients with primary headache syndromes who also suffer from mood disorders. Four-hundred and-seventy headache outpatients (170M, 300F) and 150 age- and sex-matched healthy subjects were screened using a specific questionnaire that included the Hamilton rating scales for anxiety and depression. The average scores of the Hamilton rating scales for anxiety and depression were significantly higher in headache sufferers (17.4 and 14.2, respectively) than in healthy people (6.8 and 5.7, respectively). The frequency of headache attacks, the history of headaches, and gender (women more than men) were correlated with the score of the Hamilton rating scale for both anxiety and depression. Sixteen headache patients (3.4%) achieved the DSM-IV criteria for major depression or dysthymia versus one among headache-free subjects (0.6%; OR 5.2). Patients suffering from drug-overuse and migraine with aura showed the higher odds ratios (35 and 17, respectively). These results suggest that those headache patients with long history and high frequency of headaches, or patients suffering from migraine with aura and drug-overuse might benefit from psychiatric evaluation. PMID- 10376166 TI - Platelet nitric oxide metabolites in migraine. AB - Nitric oxide (NO) is a candidate as a causative molecule in migraine. We determined nitrite, total nitrate/nitrite, and cyclic guanosine 3',5' monophosphate (cGMP) concentrations in platelets from 30 migraine without aura (MwoA) patients and 17 migraine with aura (MwA) patients. All migraine patients were studied during their migraine attacks. The control group consisted of 28 healthy volunteers. Concentrations of platelet nitrite and total nitrate/nitrite were determined using simple and sensitive nitrate/nitrite fluorometric assay techniques. High concentrations of platelet nitrite and total nitrate/nitrite were found in patients with MwoA and MwA when compared with healthy controls. High concentrations of platelet cGMP were also found in patients with MwoA and MwA. The levels of platelet total nitrate/nitrite significantly decreased in headache-free periods after treatment with oral propranolol. These findings suggest that NO is produced in platelets during migraine attacks. It may also be related to the migrainous pain and the changes in cerebral blood flow experienced during migraine attacks. These data may provide new strategies for the treatment of migraine. PMID- 10376167 TI - Prospective large-scale study of the tolerability of subcutaneous sumatriptan injection for acute treatment of migraine. AB - To investigate prospectively serious adverse events associated with sumatriptan injection, we studied 12,339 typical migraineurs for up to 12 months each. This study imitated the ordinary clinical use of sumatriptan injection except that: (a) a short, written informed consent was required, (b) there was a centralized, automated dispensing service that audited each patient's product use, (c) patients were sometimes reviewed by telephone, and (d) drug supply and medical consultation were without charge. All adverse events were recorded regardless of etiology. There were 25 fatalities during the study, none being attributable to sumatriptan injection. Of six strokes in the study, two occurred soon after treatment of a migraine attack with sumatriptan injection; whether these were migraine-related or drug-related is discussed. None of the three myocardial infarctions was due to sumatriptan injection use. We conclude that sumatriptan injection is well tolerated when used in accordance with labeling. PMID- 10376168 TI - Acute treatment of migraine attacks: efficacy and safety of a nonsteroidal anti inflammatory drug, diclofenac-potassium, in comparison to oral sumatriptan and placebo. The Diclofenac-K/Sumatriptan Migraine Study Group. AB - BACKGROUND: Migraine attacks are often treated with simple analgesics or with ergotamine-containing preparations alone or in combination with anti-emetics. Although also sometimes used to treat migraine, nonsteroidal anti-inflammatory drugs (NSAIDs) have not been systematically evaluated in controlled clinical trials, particularly in comparison with the newer drug sumatriptan. Sumatriptan is a specific migraine treatment which has recently become among the most widely prescribed acute migraine therapies. However, while effective, it has low oral bioavailability and some problematic adverse effects. Diclofenac-potassium is a potent NSAID available as a fast-acting oral tablet, which has been shown to be safe and effective in several other acute pain indications. In the clinical trial reported here, the efficacy and safety of diclofenac-potassium in the acute treatment of migraine attacks has been tested in comparison with oral sumatriptan and placebo. METHODS: Single oral doses of 50 mg and 100 mg diclofenac-potassium were compared to a single oral dose of 100 mg sumatriptan and placebo in a double blind randomized crossover trial in 156 adult patients suffering from migraine attacks, with or without aura, selected according to the International Headache Society diagnostic criteria. The primary efficacy criterion was migraine headache pain recorded on a visual analog scale at 2 h after dosing. Secondary endpoints included pain at other time points up to 8 h and the presence of accompanying symptoms (nausea, vomiting, photophobia, phonophobia). FINDINGS: Diclofenac potassium was more effective than placebo in reducing migraine headache pain at 2 h after dosing, which was the primary endpoint. Secondary analyses showed that diclofenac-potassium provided significant pain relief from 60 min after dosing and for all remaining endpoints in the 8-h observation period. Both 50 and 100 mg doses of diclofenac-potassium were similarly effective. A similar effect was shown with sumatriptan; however, significant superiority to placebo was seen only from the 90-min time point. Diclofenac-potassium was generally superior to placebo or sumatriptan in reducing accompanying symptoms, particularly nausea. Diclofenac-potassium seemed to be as well tolerated as placebo, with fewer adverse events reported than after sumatriptan treatment and with more patients assessing the overall tolerability of diclofenac-potassium better than that of sumatriptan. INTERPRETATION: Compared with placebo and the reference therapy sumatriptan, diclofenac-potassium is an effective, fast-acting, and well tolerated acute oral therapy for migraine attacks, with advantages over oral sumatriptan in terms of onset of analgesic effect, reduction of accompanying symptoms, and tolerability profile. It may therefore be useful as an alternative oral therapy for migraine attacks. PMID- 10376169 TI - Prolonged migraine aura without headache arrested by sumatriptan. A case report with further considerations. AB - The case of a 42-year-old woman with prolonged migraine visual aura without headache, whose long-lasting episodes of visual aura were successfully controlled by oral sumatriptan, is reported. Effectiveness of sumatriptan was unequivocal, since, after taking sumatriptan, duration of aura would drop from 1.5 h to approximately 20 min. This case suggests that sumatriptan may cross the blood brain barrier and block spreading depression. PMID- 10376170 TI - Migraine and ocular pain in "glaucoma suspect". AB - A relationship between glaucoma and migraine has been hypothesized by some authors, but not confirmed by others. We studied the prevalence and features of migraine and ocular pain in 460 "glaucoma suspect" patients (with ocular hypertension, but without optic disc and visual field abnormalities) and 460 controls. A higher prevalence of migraine was found in patients (13%), particularly in women (17%), than in controls (7%). At the time of the interview, migraine was still active in 68% of the patients and had decreased in the remaining 32% (prevalently those not being treated for ocular hypertension), whereas it had ceased in 52% of controls. Attacks of "ocular pain" of mild and moderate intensity were found to occur in 51% of the patients with both "glaucoma suspect" and migraine, in almost all who were not taking treatment for ocular hypertension. "Ocular pain" was time-related to the history of glaucoma. Changes in intraocular pressure may play a role in the interaction between "glaucoma suspect", migraine, and ocular pain. PMID- 10376171 TI - Efficacy definitions for migraine studies. PMID- 10376173 TI - Beta blockers: are they difficult to use in heart failure patients. PMID- 10376172 TI - Salutary effects of aspirin in coronary artery disease are not limited to its platelet inhibitory effects. PMID- 10376174 TI - Continuing medical education and industry support. AB - 1. Industry and faculty of continuing medical education programs must agree that any supported program is for scientific and educational purposes only and not to promote a company's products directly or indirectly. 2. The control of content and selection of presenters and moderators must be independent of the company and determined by the program director. 3. Faculty of continuing medical education programs must disclose any financial relationship they have with the sponsoring company. 4. Program content cannot be scripted or influenced in any way by industry. 5. Objectivity and balance must be perceived by the audience. This includes the presentation of favorable and unfavorable information and alternative treatments. 6. Incomplete data must be identified by the faculty. Oftentimes, much of what is presented at continuing medical education programs has not yet been published in peer-reviewed journals. This must be so indicated by the presenter, and it should be specified whether incomplete data is on-going research, analysis, preliminary data, or unsupported opinion. 7. Support from industry must be in the form of an educational grant with no strings attached. PMID- 10376175 TI - Apoptosis: the good and the bad--possible future therapeutic implications. PMID- 10376176 TI - The use of the New York Heart Association's classification of cardiovascular disease as part of the patient's complete Problem List. AB - This paper extols the value of combining two systems in order to improve learning, teaching, communication, patient care, and clinical research in patients with heart disease. This is accomplished by using Weed's recommendation regarding the creation of a complete Problem List and, within this context, characterizing the cardiac or vascular problem according to the recommendations of the New York Heart Association. PMID- 10376177 TI - Transesophageal echocardiography for the evaluation and management of patients with cerebral ischemia. AB - To prevent recurrent strokes and transient ischemic attacks, considerable attention is devoted to investigating the etiology of acute cerebral ischemia in the large subpopulation of patients without an easily identifiable cause. In general, transthoracic echocardiography is an insensitive tool for the evaluation of patients with cerebral ischemia, unless clinical signs and/or symptoms of cardiac disease are present. Transesophageal echocardiography (TEE), because of its increased sensitivity for aortic arch atheromata, atrial septal pathology, left atrial thrombi, and valvular abnormalities, is the preferred cardiac imaging modality, especially in young patients, older patients with hypertension or systemic atherosclerosis, and patients with prosthetic heart valves. This paper reviews the prognostic and therapeutic impact of TEE in patients with cerebral ischemia, specifically focusing on the ability of information obtained by this technique to alter patient management and improve risk stratification. PMID- 10376178 TI - The effects of metoprolol and captopril on heart rate variability in patients with idiopathic dilated cardiomyopathy. AB - BACKGROUND: The effects of treatment with captopril or metoprolol on heart rate variability (HRV) were investigated in 38 patients (29 men and 9 women) with mild to moderate symptoms of heart failure due to idiopathic dilated cardiomyopathy (DCM). HYPOTHESIS: The aim of the study was to investigate and compare the effects of the angiotensin-converting enzyme inhibitor captopril with those of the selective beta-adrenergic receptor blocker metoprolol on HRV in patients with idiopathic DCM. METHODS: Heart rate variability was analyzed in the time and frequency domains from 18th of Holter monitoring before randomized treatment was started, after 6 months of therapy, and 1 month after therapy was stopped. RESULTS: Captopril treatment increased HRV expressed as total power and low frequency power in the frequency domain. There was no change in the time domain. In the metoprolol group, there was a pronounced increase in both time- and frequency-domain indices of HRV. The increase in total power was partly maintained 1 month after therapy was stopped in both treatment groups. CONCLUSION: Treatment with captopril and metoprolol increases HRV in patients with DCM. This effect seems to be maintained for at least 1 month after therapy is stopped. The increase in HRV seems to be more pronounced with metoprolol, and the two different pharmacologic approaches may have additive effects that are of prognostic importance in patients with heart failure. PMID- 10376179 TI - QRS prolongation on the signal-averaged electrocardiogram versus ST-segment changes on the 12-lead electrocardiogram: which is the most sensitive electrocardiographic marker of myocardial ischemia? AB - BACKGROUND: ST-segment changes and QRS prolongation are electrocardiographic (ECG) markers of myocardial ischemia. HYPOTHESIS: This study was undertaken to investigate the appearance of QRS duration changes with or without concomitant ST segment changes during a typical anginal episode. METHODS: For this purpose, 126 patients underwent 12-lead surface ECG and signal-averaged electrocardiogram (SAECG) during typical anginal pain as well as at the time the patient was asymptomatic. In both periods, QRS duration and ST-segment changes were evaluated. All patients underwent cardiac catheterization. RESULTS: Of the 126 patients, 108 (86%) had coronary artery disease (CAD), whereas the remaining 18 (14%) patients had normal coronary arteriograms. During typical anginal pain, 75 of the 108 (70%) patients with CAD and 2 of the 18 (11%) patients with normal coronary arteriograms developed QRS prolongation, whereas 60 of the 108 (56%) patients with CAD and 2 of the 18 (11%) patients with normal coronary vessels developed ST-segment changes. Thus, the sensitivities of QRS prolongation measured by SAECG and of ST-segment changes on the surface ECG for the detection of myocardial ischemia were found to be 70 and 56%, respectively, (p < 0.01), whereas the specificities were both found to be 89% (p = NS). CONCLUSIONS: During typical anginal pain, QRS prolongation on the SAECG is more sensitive than are ST segment changes on the ECG for the detection of myocardial ischemia. PMID- 10376180 TI - QT dispersion plus ST-segment depression: a new predictor of restenosis after successful percutaneous transluminal coronary angioplasty. AB - BACKGROUND: ST-segment depression during exercise testing is frequently observed in the absence of restenosis after percutaneous transluminal coronary angioplasty (PTCA). HYPOTHESIS: With the goal of improving the prediction of restenosis after PTCA, we evaluated the usefulness of ST-segment depression plus QT dispersion (QTd = QTmax - QTmin) during treadmill stress test. METHODS AND RESULTS: Fifty six patients (37 men, 19 women, mean age 51 +/- 14 years) were evaluated with treadmill exercise testing and coronary angiography 7 +/- 5 months after PTCA. Treadmill test was positive in 30 patients and negative in 26 patients. At coronary angiography, restenosis was present in 16 patients with positive exercise electrocardiogram (ECG) and in 6 patients with negative exercise ECG. Fourteen patients with a positive stress test did not have restenosis. There was no difference in QTd values between groups at baseline (p > 0.05). Exercise QTd was 63 +/- 9 ms in patients with positive exercise test, 54 +/- 18 ms in patients with negative exercise test (p = 0.003), 71 +/- 13 ms in patients with restenosis, and 53 +/- 17 ms in patients without restenosis (p = 0.001). ST segment depression during the stress test determined restenosis with a sensitivity of 80% and a specificity of 58%. Sensitivity and specificity of QTd of > or = 60 ms for prediction of restenosis were 83 and 61%, respectively. When QTd of > or = 60 ms was added to ST-segment depression as a condition for positive test, the sensitivity and specificity increased to 91 and 78%, respectively. QT dispersion plus ST-segment depression had higher sensitivity and specificity than either QTd or ST-segment depression alone (p < 0.05). CONCLUSION: The addition of QTd to ST-segment depression during exercise test improves the diagnostic value and can be used as a noninvasive tool in the diagnosis of restenosis after PTCA. PMID- 10376181 TI - Paradoxical behavior of the QT interval during exercise and recovery and its relationship with cardiac memory. AB - BACKGROUND: Few studies have dealt with the behavior of the corrected (QTc) and uncorrected QT intervals during exercise and recovery. HYPOTHESIS: Based on previously published dynamics of the QT interval during treadmill testing, this study attempted to reevaluate the computer-proposed underlying mechanisms of these dynamics and to determine whether the so-called memory phenomenon could be operative in some subjects without evidence of structural heart disease. METHODS: This study included 42 unmedicated healthcare volunteers, 23 men and 19 women aged between 20 and 42 (mean 31.7) years. All had normal physical examinations, x rays, and transthoracic echocardiograms. The electrocardiograms were also normal with 12-lead QT interval dispersions of < 90 ms. RESULTS: During exercise and recovery, the behavior of the QT intervals permitted the categorization into two groups. In Group 1 (31/42; 73.8% of subjects) the uncorrected QT interval showed a biphasic pattern consisting of a gradual decrease during incremental exercise followed by a gradual increase during recovery. In contrast, the QTc interval had a triphasic pattern resulting from a slight increase during the early phase of exercise, a gradual decrease at the highest rates, and a final increase during recovery as the rate slowed to control values. In Group 2 (11/42; 36.2% of subjects) the behavior was considered as paradoxical since the uncorrected QT interval displayed in a triphasic pattern whereas the QTc interval yielded a tetraphasic pattern due to the fact that both showed a second decrease during recovery while the rate was decreasing. CONCLUSIONS: Analysis of dynamics behavior of the QTc and the uncorrected QT intervals during exercise showed that some normal subjects had a paradoxical behavior which, because of its temporal relation to the phases of exercise, could be an expression of the so-called memory phenomenon. PMID- 10376182 TI - Morning attenuation of endothelium-dependent, flow-mediated dilation in healthy young men: possible connection to morning peak of cardiac events? AB - BACKGROUND AND HYPOTHESIS: Brachial artery flow-mediated dilation (FMD), a noninvasive, widely used clinical index of endothelial function and magnitude of FMD, has been reported to be closely related to many coronary risk factors and coronary atherosclerosis. However, there has been no study that examines the diurnal change of FMD. We designed this study to reveal the diurnal variation of FMD in healthy volunteers. METHODS: We examined FMD in response to reactive hyperemia by high resolution ultrasound in 13 healthy young men (age 25-32) at four different times over the course of a day. RESULTS: Mean measures of brachial artery FMD was 4.0% at 8:00, 5.3% at 12:00, 9.7% at 17:00, and 6.9% at 21:00 hours. Flow-mediated dilation at 8:00 and at 12:00 hours was significantly lower than that at 17:00 (p < 0.05). CONCLUSIONS: These results show that endothelial function has diurnal variation and is significantly attenuated in the morning. Morning attenuation of endothelial function should be recognized in clinical research and may play an important role in the circadian variation of the occurrence of acute cardiovascular events. PMID- 10376183 TI - Fatal torsade de pointes with d,l-sotalol and low potassium. PMID- 10376184 TI - Double-orifice mitral valve. PMID- 10376185 TI - Coronary dissection and thrombosis associated with exercise testing three months after successful coronary stenting. AB - Exercise testing is commonly performed to assess the functional result of coronary revascularization procedures and is usually not associated with any complications. However, this report documents a rare case of coronary dissection and thrombosis, which resulted in an acute myocardial infarction, in a patient who underwent stress testing 3 months following successful coronary stent implantation. PMID- 10376186 TI - Metastatic germ cell tumor to the heart presenting with syncope. AB - Malignant nonseminomatous germ cell tumors (NSGCT) rarely metastasize to the heart. The first such case presenting with syncope is described. Eight previously described cases of NSGCT with intracaval metastasis to the heart are reviewed and the literature to date is discussed. Transesophageal echocardiography is the diagnostic study of choice and treatment consists primarily of platinum-based chemotherapy followed by surgical resection of residual deposits. PMID- 10376187 TI - The Botallo mystery. PMID- 10376188 TI - Clinical implications of endothelial dysfunction. PMID- 10376189 TI - Aortic stenosis. PMID- 10376190 TI - The epidemiology of triglyceride as a coronary artery disease risk factor. AB - Triglyceride (TG) has long been associated as a risk factor for coronary artery disease. A recent meta-analysis of various epidemiologic studies has confirmed this link. An important issue is to assess further the appropriate cutpoints to classify desirable TG because recent data indicate that levels < 200 mg/dl confer elevated risk. The dietary habits of present hunter-gatherer populations reveal the impact of a Westernized diet on both TG and cholesterol and suggest that a desirable TG is < 100 mg/dl. The epidemiologic and observational data in support of this concept are explored. PMID- 10376191 TI - Pathophysiology of triglyceride-rich lipoproteins in atherothrombosis: cellular aspects. AB - Elevated plasma levels of triglyceride-rich lipoproteins (TGRLP), including very low-density lipoproteins (VLDL), chylomicrons, and their remnants, are now acknowledged as risk factors for cardiovascular disease. Interactions of TGRLP with lipoprotein receptors on monocytes, macrophages, and endothelial cells may be mechanistically linked to this risk. Triglyceride-rich lipoproteins from hypertriglyceridemic (HTG) subjects have the abnormal ability to bind to low denisty lipoprotein receptors via apoE, and plasma chylomicrons from all subjects bind to a new, distinct receptor for apoB48 that is expressed specifically by monocytes, macrophages, and endothelial cells. Receptor binding and uptake of TGRLP by these cells are likely mechanisms involved in the formation of lipid filled, macrophage-derived "foam cells" of atherosclerotic lesions and for defective fibrinolysis due to endothelial dysfunction. Recognition of the atherothrombogenic potential of TGRLP may lead to improved interventions to lessen or prevent the often fatal sequelae of coronary atherosclerosis and thrombosis associated with elevated plasma triglyceride levels. PMID- 10376192 TI - Pathophysiology of triglyceride-rich lipoproteins in atherothrombosis: clinical aspects. AB - Invasive and noninvasive arterial imaging are important techniques used to study atherosclerosis and, specifically, to evaluate the atherogenecity of triglyceride rich lipoproteins (TRL). Serial coronary angiography trials show significant benefit from lowering low-density lipoprotein cholesterol (LDL-C) which serves to retard lesion progression. Even with aggressive LDL-C reduction, however, up to half of patients demonstrate continued progression of atherosclerosis. Angiographic studies reveal that lowering LDL-C has the most impact on severe lesions, those > or = 50% diameter stenosis, whereas TRL (and their apolipoprotein markers) have been identified as a driving factor behind progression of mild-to-moderate lesions < 50% diameter stenosis. Quantitative coronary angiography (QCA) has demonstrated that progression of mild-to-moderate lesions are among the most significant predictors of clinical coronary events, and that lowering TRL reduces progression of coronary artery disease to the same degree as the lowering of LDL-C. PMID- 10376193 TI - Measurement of triglyceride-rich lipoproteins by nuclear magnetic resonance spectroscopy. AB - Nuclear magnetic resonance (NMR) spectroscopy is being used to determine the concentrations of very low-density lipoprotein (VLDL), low-density lipoprotein (LDL), and high-density lipoprotein (HDL) subclasses of different size. These subclasses have unequal associations with coronary heart disease. Nuclear magnetic resonance distinguishes among the subclasses on the basis of slight differences in the spectral properties of the lipids carried within the particles, which vary according to the diameter of the phospholipid shell. Studies using NMR spectroscopy have shown that individuals with elevated triglycerides are likely to have higher-risk lipoprotein subclass profiles. Triglyceride-rich lipoproteins drive the metabolic reactions that produce LDL of abnormal size and cholesterol content. The quantities of these abnormal LDL particles and the associated risk of coronary heart disease are underestimated by conventional cholesterol measurements. Nuclear magnetic resonance spectroscopy measures lipoprotein subclasses directly and efficiently, and produces information that may improve the assessment and management of cardiovascular disease risk. PMID- 10376194 TI - Postprandial triglyceride metabolism in diabetes mellitus. AB - Individuals with diabetes have a two to four times higher risk of cardiovascular morbidity and mortality than nondiabetics. Patients with both type 1 and type 2 diabetes share a similar risk. Studies in individuals with type 1 diabetes have shown a decreased clearance of postprandial triglyceride-rich lipoprotein particles of abnormal composition. Particles isolated from diabetic individuals show abnormal composition and an increased tendency to cause cholesteryl ester accumulation in macrophages and are therefore potentially atherogenic. Various interventions may alter these abnormalities and improve the atherosclerotic risk. These include adopting a high-carbohydrate diet over a high monounsaturated diet, improving glycemic control, infusing insulin intraperitoneally, and using pharmacologic therapies such as the statins. PMID- 10376195 TI - Brachial artery ultrasound: a noninvasive tool in the assessment of triglyceride rich lipoproteins. AB - In recent years, endothelial dysfunction has been identified as an early feature of atherosclerosis. Endothelial function can be measured noninvasively by using brachial artery ultrasound. A variety of factors associated with atherosclerosis also impair endothelial function. Some of these factors are lipoproteins such as various forms of low-density lipoproteins, postprandial chylomicron remnants, fasting triglyceride-rich particles, and free fatty acids. A high-fat diet also has an adverse effect on endothelial function. Several interventions can improve endothelial function and, at the same time, reduce cardiovascular events. Measuring endothelial function may eventually serve as a useful index to determine an individual's risk for coronary artery disease. PMID- 10376196 TI - Nonpharmacologic treatment of hypertriglyceridemia: focus on fish oils. AB - Early studies in Greenland Eskimos stimulated interest in evaluating the effect of Omega-3 fatty acids on coronary artery disease. Subsequent studies showed a significant decrease in triglyceride levels in patients receiving high doses of fish oil containing DHA and EPA. Slight increases in LDL were also observed in patients receiving fish oil supplements. These studies have also shown a dose response effect which persists as long as supplementation continues. Later trials, specifically the Diet and Reinfarction Trial and the Indian Experiment of Infarct Survival, have demonstrated a reduction in cardiac death rates and in the incidence of cardiac symptoms in patients receiving fish oil. PMID- 10376197 TI - Pharmacologic management of triglycerides. AB - Currently available cholesterol-lowering pharmacologic agents have been studied for their effect on reducing triglyceride levels. The fibrates increase lipoprotein lipase activity, thereby decreasing the size of triglyceride-rich particles. High doses of niacin can produce decreases in very low-density lipoprotein (VLDL) levels, triglyceride-rich particles, and low-density lipoprotein (LDL) by inhibiting hepatic lipoprotein synthesis. By increasing LDL receptor activity, the statins increase the removal rate of triglyceride-rich particles. Each class of agents produces various degrees of triglyceride lowering, depending on the existing baseline level and other factors. Patients with elevated LDL who are also hypertriglyceridemic should receive statins as first-line therapy. Niacin may be used as an alternative first-line agent in patients with low LDL elevations. Combination therapy using other agents may be indicated depending on the patient's levels of triglycerides and LDL. PMID- 10376198 TI - The effect of early postinfarction revascularization of asymptomatic patients on left ventricular remodeling. AB - BACKGROUND: Patients with angina after a Q-wave myocardial infarction benefit from elective revascularization, but it is not known whether asymptomatic patients, including those with a totally occluded infarct-related artery, improve after revascularization. OBJECTIVE: To determine the effect of early postinfarction revascularization of asymptomatic patients on left ventricular remodeling. METHODS: We prospectively studied 31 consecutive asymptomatic patients (aged 57 +/- 2 years, 24 with anterior infarcts) after Q-wave myocardial infarction with > or = 70% stenosis of the infarct-related artery (IRA) who underwent early elective revascularization (days 4-10 after myocardial infarction). Group I consisted in patients with a totally occluded IRA (n = 10), and group II consisted in patients with a patent, though stenosed, IRA (n = 21). Resting echocardiography and low-dose dobutamine echocardiography were performed at baseline (day 3 +/- 1), and rest echocardiography was repeated after an 8-week follow-up. Significant myocardial viability was defined as > or = 2 wall segments improved (in a 16-segment model of left ventricle) versus baseline, and significant functional recovery as > or = 2 segments improved versus baseline on follow-up examination. Left ventricular end-systolic volume indices (ESVI) and end-diastolic volume indices and ejection fractions were measured by using a modified version of Simpson's rule (using apical two-chamber and four-chamber views). RESULTS: The left ventricular ESVI of patients in group I had decreased by 4.2 +/- 1.9 ml/m2, whereas for patients in group II the left ventricular ESVI had increased by 4.2 +/- 1.7 ml/m2 (P = 0.006). Similarly, the left ventricular end-diastolic volume index had decreased by 0.7 +/- 2.4 ml/m2 versus baseline at follow-up for patients in group I and increased by 7.8 +/- 2.1 ml/m2 for patients in group II (P = 0.02). The left ventricular ejection fraction increased by 7.3 +/- 3% for patients in group I and decreased by 0.4 +/- 2% for patients in group II (P = 0.04). CONCLUSION: There is less global left ventricular remodeling, a potentially deleterious process, after elective revascularization early after Q wave myocardial infarction in asymptomatic patients who had had a totally occluded IRA before revascularization than there is in patients who had already had a patent, though stenosed, IRA before revascularization. These results suggest that restoration of patency of IRA after a Q-wave myocardial infarction is beneficial even for asymptomatic patients. PMID- 10376199 TI - Coronary risk factors influence plaque morphology in patients with unstable angina. AB - BACKGROUND: The risk of plaque disruption and subsequent thrombosis in patients with unstable angina depends on the plaque type and size. DESIGN: Intravascular ultrasound (IVUS) was employed to illustrate the correlation between risk factors and plaque morphology in patients with unstable angina. METHODS: In a prospective study of 60 of 95 patients consecutively admitted with unstable angina [41 men, aged 61.2 +/- 8.1 years (mean +/- SD)], qualitative (soft and hard plaque, thrombus, calcification, eccentricity, adaptive and constrictive remodeling) and quantitative [lumen, external elastic membrane (EEM) and plaque cross-sectional area (CSA) and plaque burden] IVUS data relating to the target lesion, and proximal and distal reference segments were analyzed and correlated with risk factors. Univariate and multivariate nominal logistic regression analyses and analyses of variance were used to determine the independent predictors for IVUS morphology. RESULTS: For plaque composition univariate analysis showed a younger age (< 60 years) to be a predictor for adaptive remodeling (P = 0.019), and an older age to be a predictor for constrictive remodeling (P = 0.021). Hypercholesterolemia, smoking and sex were associated with a higher frequency of thrombus (P = 0.044, 0.038 and 0.043, respectively). Multivariate analyses revealed that only younger and older ages were independent predictors for adaptive and constrictive remodeling (P = 0.039 and P = 0.045). For plaque size, univariate and multivariate analyses demonstrated that diabetes mellitus and hypercholesterolemia were independent predictors for greater plaque (13.5 +/- 5.72 versus 10.17 +/- 4.6 mm2, P = 0.015, for diabetic versus non-diabetic patients; 12.0 +/- 5.35 versus 9.03 +/- 3.76 mm2, P = 0.010, for hypercholesterolemic versus normocholesterolemic patients) and EEM CSA (17.16 +/- 5.81 versus 14.3 +/- 5.1 mm2, P = 0.033, for diabetic versus non-diabetic patients; 16.57 +/- 5.49 versus 12.25 +/- 3.8 mm2, P = 0.001, for hypercholesterolemic versus normocholesterolemic patients) at the target lesion. Hypercholesterolemia was associated with significantly greater plaque and EEM CSA in both proximal and distal reference segments. CONCLUSIONS: Multivariate analyses indicated that age, diabetes and hypercholesterolemia are independent predictors for plaque morphology in patients with unstable angina. PMID- 10376200 TI - Clinical implications of circulating soluble Fas and Fas ligand in patients with acute myocardial infarction. AB - BACKGROUND: Apoptotic cell death is the major form of myocardial damage produced by coronary ischemic events. OBJECTIVE: To assess whether circulating levels of soluble Fas (sFas), an inhibitor of apoptosis, and sFas ligand, an inducer of apoptosis, in patients with coronary artery disease are greater than normal. METHODS: Forty-seven patients [acute myocardial infarction (AMI) in 17, old myocardial infarction (OMI) in 15, stable angina in 15] and 10 normal control subjects participated in this study. Serum levels of sFas and sFas ligand in all patients were measured, and cardiac catheterizations were performed. RESULTS: Serum levels of sFas were greater than normal only in patients with AMI (4.6 +/- 1.6 ng/ml); the levels were significantly higher than those in patients with OMI (2.1 +/- 0.6 ng/ml) and stable angina (2.2 +/- 0.5 ng/ml), and in normal subjects (2.0 +/- 0.6 ng/ml; P < 0.0001). However, there was no difference among serum levels of sFas ligand for all groups. For patients with AMI, there was no significant correlation between serum levels of sFas and peak levels both of plasma creatine phosphokinase and of plasma myosin light chain type I as clinical indexes of infarct size. However, there were significant correlations between serum levels of sFas and both pulmonary artery wedge pressure (r = 0.767, P = 0.0003) and left ventricular end-diastolic pressure (r = 0.629, P = 0.03). CONCLUSIONS: Circulating sFas increases in concentration in relation to the severity of hemodynamic conditions in patients with AMI, but it is independent from size of infarct. Therefore, circulating sFas could play an important role as the marker of pathophysiologic conditions associated with cardiomyocyte apoptosis in AMI. PMID- 10376201 TI - Variant angina is not associated with angiotensin I converting enzyme gene polymorphism but rather with smoking. AB - BACKGROUND: Angiotensin converting enzyme (ACE) perhaps plays roles in regulating coronary vasomotor tone by producing angiotensin II and degrading bradykinin. OBJECTIVES: We sought to investigate the role of ACE gene polymorphism in the pathogenesis of variant angina and to compare it with that of other clinical risk factors for male patients with variant angina and age-matched and sex-matched control subjects. METHODS: We studied 78 male patients with variant angina who exhibited spontaneous or provoked coronary spasms during coronary angiography and compared prevalences of ACE gene genotype (deletion D and insertion I) and other risk factors between this group of patients with variant angina and age-matched and sex-matched control subjects whose angiograms were normal and in whom the ergonovine test did not cause spasms (n = 80). RESULTS: Smokers were more prevalent in the group of patients with variant angina (P < 0.05). Genotype and allele prevalences of the group of patients with variant angina (0.14, 0.53 and 0.33 for DD, DI and II and 0.41 and 0.59 for D and I, respectively) were no different from those of the control group (0.16, 0.49 and 0.35 for DD, DI and II and 0.40 and 0.60 for D and I). Multiple logistic regression analysis showed that smoking was a significant risk factor for variant angina (odds ratio 2.61, 95% confidence interval 1.03-6.66) whereas ACE genotype was not. CONCLUSIONS: Variant angina is associated with an environmental factor, such as smoking, rather than a genetic factor, such as ACE gene polymorphism. PMID- 10376202 TI - The effects of dipyridamole stress test on plasma levels of soluble adhesion molecules intracellular adhesion molecule-1, vascular cell adhesion molecule-1, E selectin and L-selectin in patients with ischemic heart disease and patients with syndrome X. AB - BACKGROUND: Adhesion of activated leukocytes to the endothelium as a result of myocardial ischemia/reperfusion has been shown to be involved in the development of tissue injury. Leukocyte adhesion to the endothelium occurs via adhesion molecules expressed on the surface of both cell types. Upon cell activation these proteins may be released into the circulation and measured in a soluble form. AIM: To verify whether the dipyridamole stress test, performed in patients with ischemic heart disease (IHD) and in patients with syndrome X, modifies plasma levels of the soluble adhesion molecules vascular cell adhesion molecule-1 (VCAM 1), intracellular adhesion molecule-1 (ICAM-1), E-selectin and L-selectin. METHODS: Plasma levels of the soluble endothelial adhesion molecules ICAM-1, VCAM 1 and E-selectin, as well as of the soluble leukocyte adhesion molecule L selectin, were measured in venous blood samples taken before and 7 min after administration of dipyridamole in patients with IHD, patients with syndrome X and healthy individuals. Myocardial perfusion was evaluated using single photon emission tomography. The plasma levels of soluble VCAM-1, ICAM-1, E-selectin and L-selectin were all measured using enzyme-linked immunosorbent assays. RESULTS: After infusion of dipyridamole, plasma levels of ICAM-1 increased significantly in patients with IHD, whereas they remained unchanged in patients with syndrome X and in the control group. In patients with IHD, the initial plasma levels of VCAM 1, E-selectin and L-selectin, before administration of dipyridamole, were higher than those observed in patients with syndrome X and than those in the control group. Plasma levels of soluble VCAM-1, E-selectin and L-selectin decreased significantly in patients with IHD following the dipyridamole stress test, whereas they remained unchanged in patients with syndrome X, and in the control group. CONCLUSION: In patients with IHD, administration of dipyridamole induces myocardial ischemia resulting in modification of plasma levels of the soluble adhesion molecules. PMID- 10376203 TI - Endothelial function and atherosclerosis. AB - Under normal conditions, the vascular endothelium exerts vasodilator, antithrombotic, and growth-inhibiting effects on the vessel wall. In the setting of atherosclerosis, endothelial dysfunction contributes to vasoconstriction, thrombosis, and growth of vascular smooth muscle. This article reviews the important contributors to endothelial function both in health and in the diseased state of atherosclerosis. Therapeutic options for improvement of endothelial function will also be summarized. PMID- 10376204 TI - Blood pressure profile in patients with microvascular angina. AB - Normotensive patients with microvascular angina exhibit increased diastolic blood pressure and blood pressure loads during daily activities and decreased diurnal variation of systolic blood pressure, compared with age- and sex-matched normotensive controls. The abnormal blood pressure profile could play a role in the pathogenesis of microvascular angina. PMID- 10376205 TI - Magnetic resonance imaging of the coronary arteries: anatomy of the coronary arteries and veins in three-dimensional imaging. AB - Magnetic resonance imaging of coronary arteries will visualize, besides the arteries, the myocardium, blood in the cavities and cardiac veins. This will hamper the application of projectional visualization techniques such as those used in conventional coronary angiography. Volume rendering, a different visualization technique, can be used to create a three-dimensional impression of a magnetic resonance data set on a two-dimensional surface. In this article, we will review the volume-rendering technique and anatomy of the coronary arteries and veins in the obtained images. Also we will discuss the relation between arteries and veins and the possible sites of confusion. PMID- 10376206 TI - Bibliography. Current world literature. PMID- 10376207 TI - Cloning and sequencing of the gilthead sea bream estrogen receptor cDNA. AB - We report here the complete nucleotide sequence of a cDNA clone containing the full-coding sequence of the Sparus aurata estrogen receptor (ER) isolated from an expression library prepared from gilthead sea bream liver poly A+ RNA. The library was screened using a single strand rainbow trout ER cDNA probe, corresponding to the C-D domain. The cDNA sequence containing an insert of 2369 nucleotides was found to encode a protein of 579 amino acids. The 5'- and 3' untranslated regions of the message are 186 and 392 nucleotides long, respectively. The gilthead sea bream ER shows the higher homology with the ER of another perciform, Chrysophrys major (93%), moderate to high homology with Oreocromis aureus (78%) medaka (77%) and rainbow trout (70.7%) ERs and lower homology with japanese eel (45%), amphibian (47%), avian (48.5%) and mammalian (47-47.5%) ERs. The sequence homologies and phylogenetic analysis of the various ERs suggest that gilthead sea bream ER should be considered as a ER alpha-like. PMID- 10376208 TI - Guinea pig D-amino-acid oxidase cDNA and phylogenetic position. AB - The nucleotide sequence of cDNA that encodes guinea pig D-amino-acid oxidase (DAO) was determined. The cDNA consisted of 1,399 nucleotides and a poly(A) tail. The cDNA encodes 347 amino acid residues. In contrast to the hamster, rat, and mouse DAOs, guinea pig DAO had the 25th amino acid residue. The homology in amino acid sequences between the guinea pig DAO and the rodent DAOs was not high in comparison to the homology in amino acid sequences between the guinea pig DAO and DAOs of humans, pigs and rabbits. The phylogenetic position of the guinea pig varied depending on the source of sequences (amino acids or nucleotides) and the methods of phylogenetic tree construction. These results suggest that the guinea pig is not a simple rodent. PMID- 10376209 TI - Nucleotide and peptide sequences of the open reading frame encoding a truncated toxin A gene of Clostridium difficile strain CCUG 20309. AB - The open reading frame encoding a putative truncated toxin A gene of Clostridium difficile in strain CCUG 20309 (ATCC 8864), a strain that produces toxin B but not toxin A, was sequenced by cycle sequencing method. The coding region contains 2097 base pairs and has a GC content of 26.4%. The deduced polypeptide is 50 kDa and is generally hydrophilic. Although strain CCUG 20309 of C. difficile was reported not to produce toxin A, it is enterotoxic, an inherent property of toxin A in pathogenic strains of C. difficile. It therefore remains to be determined whether a truncated form of toxin A could actually be expressed from the open reading frame by this strain of C. difficile. PMID- 10376210 TI - Molecular cloning and sequence analysis of bubaline lactoferrin promoter. AB - The proximal promoter for bubaline lactoferin-encoding gene has been isolated, cloned and sequenced. A 468 bp fragment of the 5' flanking region of the lactoferrin gene was PCR amplified and cloned into pUC18 vector. Sequence analysis of the amplified fragment revealed the presence of one TATA box, one TATA like element, two GC boxes and one motif resembling cAMP response element (CRE) in this region. Bubaline lactoferrin promoter shares 93%, 53%, 52% and 48% homology with cattle, pig, mouse and human lactoferrin 5' flanking region, respectively. PMID- 10376211 TI - cDNA cloning and sequence analysis of bubaline growth hormone. AB - The cDNA for Bubalus bubalis growth hormone (GH) has been cloned and sequence determined through RT-PCR approach. The nucleotide sequence of bubaline GH cDNA was in a single reading frame coding for a protein of 191 residues comprising a putative signal sequence of 27 amino acids. Homology comparison of the sequence with other mammalian GH cDNAs showed a very high degree of evolutionary conservation. Bubaline GH sequence shared a homology of 99.5%, 99.5%, 98.6%, 87.6% and 61.9% with that of ovine, caprine, bovine, porcine and human, respectively at amino acid level. PMID- 10376213 TI - Caenorhabditis elegans contains structural homologs of human prk and plk. AB - We and others have recently reported cloning and characterization of human prk and plk, members of the polo family of protein serine/threonine kinases that includes the budding yeast cdc5 and Drosophila melanoganster polo. The cdc5 gene is essential for cell cycle progression through mitosis and controls adaptation to the yeast DNA damage checkpoint. Here we report the identification of two new cdc5 homologs from Ceanorhabditis elegans, named plc1 and plc2. The deduced amino acid sequences of Plc1 and Plc2 share strong homology with both human Prk and Plk. plc1 and plc2 genes are closely linked on chromosome III and share 40% residue identity, suggesting that gene duplication followed by independent evolution gives rise to multiple polo homologous genes within a species. Similar to polo family members in other species, two distinct domains are present in Plc1 and Plc2 with the N-terminal half being the putative kinase domain. Interestingly, Plc2, unlike Plc1, contains a less conserved polo box within the C terminal half of the protein, suggesting a functional division between these two kinases. PMID- 10376212 TI - Molecular cloning and sequence analysis of the cDNA encoding beta-lactoglobulin in Bubalus bubalis. AB - The cDNA for bubaline beta-lactoglobulin (beta lg) has been cloned through RT-PCR approach and sequenced. Sequence data showed a single open reading frame coding for a protein of 180 amino acids with a signal sequence of 18 amino acid residues. Comparison with other ruminant beta lg sequences revealed a high homology indicating the protein to be conserved through evolution. The degree of homology, at amino acid level, is 96.1%, 95.6%, 93.9% and 63.7% with goat, sheep, cow and pig, respectively. PMID- 10376214 TI - A pseudoautosomal boundary-like element adjacent to the SSAV1 locus at 18q21. AB - Pseudoautosomal boundary-like (PABL) elements have been found at transition sites between genomic regions with different GC-contents. A new PAB related sequence was found immediately adjacent to the S71 provirus on human chromosome 18q21.1-2 (officially designated the SSAV1 locus). The S71 PABL element was full-length as defined by comparison with elements identified at the pseudoautosomal boundaries of the sex chromosomes and in the MHC region. The 3'-ends of all PABL elements showed significant homology to functional CpG-islands, indicating that this similarity is a new common feature of PABL elements. PMID- 10376215 TI - A comparative study of rat and human Tmp21 (p23) reveals the pseudogene-like features of human Tmp21-II. AB - Tmp21 (p23) is involved in biosynthetic transport from the endoplasmic reticulum to the Golgi complex. We have recently characterized two cDNA-variants of human Tmp21, the Tmp21-isoforms-I and -II. Because of the lack of cDNA sequence data and protein expression data, it was not clear if Tmp21-II encodes a functional Tmp21-protein. Here we describe the cloning of the full length human Tmp21-II transcript. The putative open reading frame of Tmp21-II contains a reading frame jump and a nonsense mutation in comparison to all other Tmp21-I (p23) members. Our data indicate that hum-Tmp21-II is transcribed, but not translated. We conclude that Tmp21-II cDNA derives from a neutral pseudogene, which originates from a duplication event of the human Tmp21-isoform-I. PMID- 10376217 TI - Nucleotide sequence of equine caspase-1 cDNA. AB - Caspases are a family of cysteine proteases which have important roles in activation of cytokines and in apoptosis. Caspase-1, or interleukin-1 beta converting enzyme (ICE), promotes maturation of interleukin-1 beta (IL-1 beta) and interleukin-18 (IL-18) by proteolytic cleavage of precursor forms to generate biologically active peptides. We report the cloning and sequencing of equine caspase-1 cDNA. Equine caspase-1 is 405 amino acids in length and has 72% and 63% identity to human and mouse caspase-1, respectively, at the amino acid level. Sites of proteolytic cleavage and catalytic activity as identified in human caspase-1, are conserved. PMID- 10376216 TI - Unique 5'-end of a Na(+)-K(+)-2Cl- cotransporter-like mRNA expressed in rat skeletal muscle. AB - Functional evidence presented by others indicates that rat slow-twitch skeletal muscle lacks typical Na(+)-K(+)-2Cl- cotransporter activity, as determined by loop diuretic-sensitive potassium transport. This report presents a unique 5' mRNA sequence of a Na(+)-K(+)-2Cl- cotransporter-like molecule expressed in the rat soleus muscle and the deduced N-terminus of the protein. In addition to its unique 5' mRNA sequence, the coding region of the N-terminus is quite short compared with other known Na(+)-K(+)-2Cl- cotransporters. Nonetheless, the mRNA possesses conserved cotransporter-like membrane spanning domains, though one domains corresponding to a reported exon is divergent. Therefore, it appears that skeletal muscle does express a Na(+)-K(+)-2Cl- cotransporter-like mRNA that may code for a protein with atypical Na(+)-K(+)-2Cl- cotransporter properties. PMID- 10376218 TI - The characterisation of a cervine immunoregulatory cytokine, interleukin 12. AB - The cloning, sequencing, and production of cervine interleukin-12 is described. The cervine IL-12 p35 subunit coding sequence is 666 bp long and has highest homology to bovine p35 (94%), followed by human (79%), then murine (57%). The cDNA codes for a 221 aa long protein with predicted molecular weight of 24,902 Da. The cervine p40 subunit has a coding sequence of 984 bp and shows 96% homology to bovine, 85% homology to human, and 65% homology to murine p40 respectively. Cervine p40 cDNA codes for a 327 aa long protein with a predicted molecular weight of 37,461. Both subunits were inserted into a recombinant baculovirus that was then used to produce cervine IL-12 in Trichoplusia ni cells. Interleukin-12 was secreted into the culture medium and was biologically active as measured by proliferation of mitogen sensitised peripheral blood lymphocytes and the induction of interferon-gamma transcription in peripheral blood lymphocytes. PMID- 10376219 TI - Practice nurses' perceptions of services for clients with psychological problems in primary care. AB - Over a third of people presenting in primary care in the United Kingdom (UK) have a mental health problem causing some degree of disruption in their lives. Approximately 90% of these are treated and managed by primary care staff without any support from mental health services. Following the White Paper published by the UK Department of Health in 1997 (Department of Health, 1997. The New NHS- Modern and Dependable, HMSO, London), the influence of primary care both in the commissioning and provision of mental health services is likely to increase. By far the largest professional group currently involved in mental health in primary care are practice nurses. Although their numbers have increased dramatically during this decade, little is known of the work they do or of their perceptions of it. The present questionnaire-based study sought to elicit the types of mental health problems encountered by practice nurses in primary care, the interventions they provide and the skills they utilise. The data indicates that practice nurses care for people with a wide variety of mental health problems ranging from mild to severe. Many feel unprepared for this type of work and are reluctant to get too involved with clients in case they uncover problems they are not able to cope with. Lack of access to appropriate educational support is identified as the main problem currently faced by practice nurses alongside poor inter-professional relationships with mental health personnel. This paper discusses ways of meeting the needs of practice nurses and of improving collaboration in primary care settings. PMID- 10376220 TI - Qualified nurses' lived experience of violence perpetrated by individuals suffering from enduring mental health problems: a hermeneutic study. AB - The incidence of violence within the National Health Service is rising and attention to the issue is increasing. Due to the ramifications in terms of physical, psychological and economic cost, the need to understand all the dynamics and variables involved in violence becomes evident. If nurses can provide care that reduces the frequency, intensity and negative consequences of violence, then clients, nurses and the organisation all benefit. This study attempted to discover the lived experience of nurses who experience violence perpetrated by individuals suffering from enduring mental health problems. It adopted a hermeneutic, phenomenological, method and produced an emerging theory comprised of the three key themes; Personal construct of violence, Feeling equipped and Feeling supported. Furthermore, the author suggests relationships between exposure to violent incidents and the nurse's ability to deal with the incidents therapeutically and how formal support systems for nurses influence this relationship. Strategic plans that are concerned with caring for violent individuals need to consider this reciprocity, as staff who feel well supported may well have a substantial impact on the quality of care offered to these people. PMID- 10376221 TI - A review of spirituality as applied to nursing. AB - In this paper a review of spirituality as applied to nursing is carried out. In doing so, it is shown that the holistic understanding of spirituality has been derived almost exclusively from the Christian theological tradition. Whilst recognizing the importance of this tradition, the relatively unknown element, that is, the biological basis of spirituality as advanced by empirical research on spiritual awareness is brought to the debate in this review. Following the review, an operational definition of spirituality embracing its biological roots is provided to highlight its significance to nursing. PMID- 10376222 TI - Enrolled nurses and the professionalisation of nursing: a comparison of nurse education and skill-mix in Australia and the UK. AB - In the UK prior to 1989 two levels of nurse were trained: first level, or 'Registered Nurses' (RNs), and second Level, or 'Enrolled Nurses' (ENs). In 1989 changes to nurse education driven by 'Project 2000' marked the end of EN training: nurse education moved into the higher education sector and a single type of RN education replaced the original split-level training. Yet in Australia, where RN training has followed a similar path into higher education, the split level training of ENs and RNs has been maintained. The reasons for this difference in approach to ENs are investigated and discussed. The paper goes on to explore the implications and possible outcomes of the two different approaches in terms of the professionalisation of nursing and skill-mix in the health care workforce. Now that some UK nursing bodies are pressing for a degree-led profession, it is suggested that the Australian model may have an advantage, as concerns are being raised that English nurses may 'price themselves out of the market', with the nursing role being encroached upon by non-nurse Health Care Assistants. PMID- 10376223 TI - Evaluating the benefits of a clinical ladder for nursing staff: an international review. AB - This paper reviews the use of clinical ladders for nursing staff and examines the extent to which their claimed benefits have been realised, on the basis of available published research-based evaluation. A 'clinical ladder' is a grading structure which facilitates career progression and associated differentation of pay by defining different levels of clinical practice. The review examines publications from the United States, New Zealand, Australia and the United Kingdom. It notes that the vast majority of publications are descriptive 'this is how we did it' articles which do not provide any evaluation. The relatively few published evaluations are assessed, and it is concluded that the evidence base for supporting the claimed benefits of the use of clinical ladders is fragmented. Methods of evaluation have varied between studies, and in some cases fall short of that required for objective analysis. PMID- 10376224 TI - Evaluation of a nurse-run asthma school. AB - Important aims of the study were to investigate whether an educational program (the 'Asthma School') directed by a nurse led to improved knowledge of the disease, to improved self-medication and self-management and to improved, self rated, functional status. A total of 32 patients (6 males, 26 females, mean age 43 years) was included. The following methods were used to collect the data before and one year after the Asthma School was completed; two study-specific questionnaires for collecting demographic data and measuring different aspects of the patients' knowledge of the disease and its treatment, monthly diary cards, lung-function tests (FEV1) and the Sickness Impact Profile (SIP) questionnaire. The main results of the study were an improved knowledge of the disease and its treatment, better self-management, i.e. more frequent use of the peak expiratory flow meter (PEF-meter) and use of inhaled bronchodilators on an as-required basis, fewer patients on sick-leave and a better, self-rated, physical health status. However, in spite of these encouraging results, the lungfunction was found to be unaffected, no pronounced changes in the use of asthma drugs could be found and the patients' need for medical care remained the same. PMID- 10376225 TI - Medication compliance and older people: a review of the literature. AB - The purpose of this paper is to critically review a select body of literature pertaining to medication compliance among older people. As there is a vast amount of literature on the topic, this review is selective to include (1) a critique of the more commonly cited compliance theories, (2) an analysis of the key variables influencing medication compliance among older people and (3) a critical evaluation of the literature which examines these phenomena. In addition, studies, which explore the benefits of Self-Administration of Medication Schemes, are the subject of particular scrutiny, by virtue of the fact that they address not only the education component but also the behavioural component of a well organised patient education programme. The review is limited to material published in English since 1980 with the exception of the literature on compliance theories, which originated in previous decades. Key search terms including medication, compliance, older people, self-medication and education were used to derive the relevant material from the Medline and CINAHL databases. The literature was then critically reviewed using the criteria identified by Roe (1993) [Roe, B., 1993. Undertaking a critical review of the literature. Nurse Researcher 1(1), 35-46.] which emphasises the need for clarity in key areas such as research design, sample selection, research methods, results, discussion and conclusions. Empirical research which met this criteria was included in the review and in the main, this was found in academic rather than professional journals. The review concludes with a summary of the main points and a discussion of the implications for nursing practice, education and research. PMID- 10376226 TI - Occupational mental health: a study of work-related depression among nurses in the Caribbean. AB - This study addresses issues of occupational mental health among nurses in the Caribbean. A linear model linking role, work and social factors, stress, burnout, depression, absenteeism and turnover intention guides the research. Data were collected from 119 nurses working for major hospitals located in St. Vincent and Trinidad & Tobago using a field survey. Psychometrically sound instruments with proven cross-cultural validity were utilized in the questionnaire. Descriptive statistics, correlations, and path analysis were used to analyze the data. The results indicated fairly strong support for the proposed model which is tested for the first time among a Caribbean population. Role conflict, role overload and social support predicted stress, which along with social support predicted burnout. Burnout was the sole predictor of depression which in turn predicted both absenteeism and turnover intention. Implications of these findings for research and practice are discussed. PMID- 10376227 TI - A review of the impact of pre-operative education on recovery from surgery. AB - Seminal research by Boore (Boore, J., 1978. Prescription for Recovery. RCN, London) and Hayward (Hayward, J., 1975. Information--a Prescription against Pain, RCN, London) demonstrated that pre-operative information reduces post-operative stress, pain and anxiety in general surgical patients. Both studies used experimental designs and the results are frequently cited in the UK nursing literature. However, they are now more than twenty years old and surgical practices, patterns of hospitalisation and nursing as well as patients' knowledge and expectations have changed enormously. Other more recent studies have sought to evaluate the impact of pre-operative education on post-operative recovery. This paper reviews the research published in this field since 1985. It focuses on studies in which an experimental design was used and considers the types of educational intervention employed and the impact on patient outcomes. A sequel paper will evaluate the research methods used in the light of current practice in the design and reporting of randomised controlled clinical trials. PMID- 10376228 TI - Pre-operative education--a review of the research design. AB - Trials on the effectiveness of providing people with education before surgery have suggested a beneficial effect. Although this has been demonstrated through meta-analyses, the results from individual studies have not always shown significant differences between control and experimental groups. This could be a product of the design of the studies. Hence this paper examines the research literature on pre-operative education in the light of modern standards in design and reporting of randomised controlled trials. Patient assignment, blinding of participants and researchers, follow-up procedures and statistical analyses are considered. The theoretical framework underpinning this body of research and ethical issues are discussed. Although there are a relatively large number of studies on pre-operative education the conclusion is that there is considerable room for improvement in trial design as a basis for promoting evidence based nursing. PMID- 10376229 TI - Diagnostic errors of primary care screens for depression and panic disorder. AB - OBJECTIVE: As the health care reimbursement system has changed, brief screens for detecting mental disorders in primary care have been developed. These efforts have faced the formidable task of identifying patients with mental disorders, while at the same time minimizing the number of misclassified cases. Here we consider the balance between sensitivity and positive predictive value. Primary care patients with false positive and false negative results on screens for depression and panic disorder are compared with regard to comorbidity and functional impairment. METHODS: This was a cross-sectional psychometric study. The study sample included 1001 primary care patients from the Department of Internal Medicine at Kaiser Permanente in Oakland, California. The Symptom-Driven Diagnostic System for Primary Care (SDDS-PC) screens and Sheehan Disability Scale were completed by the subjects. The SDDS-PC diagnostic interviews were administered to all subjects. RESULTS: Patients with false positive results on the panic disorder screen did not differ from patients with false negatives results with regard to rates of other psychiatric disorders, functional impairment, or mental health service utilization. In contrast, patients with false negative depression screen results had significantly more psychiatric disorders and functional impairment than those with false positive depression results. CONCLUSIONS: A substantial number of patients with either false positive or false negative screen results met diagnostic criteria for other mental disorders. Given the nominal burden of follow-up assessments for patients with positive screens, these data suggest that erring on the side of sensitivity may have been preferable when algorithms for these screens were selected. PMID- 10376230 TI - Psychiatric assessment of candidates for bone marrow transplantation: a psychodynamically-oriented approach. AB - OBJECTIVE: To seek possible relationships between psychological factors and survival after an intensive medical therapy, using bone marrow transplantation as a model. METHOD: Candidates for bone marrow transplantation underwent two to three psychodynamically-oriented psychiatric interviews that explored family functioning ("F"), individual psychological maturity ("I"), and the capacity to form and communicate a mature psychological construct of the transplant ("T") process. The results were recorded in a semiquantitative manner, assigning a possible score of 1 to 3 for each parameter, for a possible total of 3 to 9 (the "F.I.T." assessment). Survival after the transplant was analyzed retrospectively in relation to the F.I.T. assessment. RESULTS: In a series of 112 candidates interviewed prior to transplant, those with the lowest F.I.T. assessment tended not to survive as long. By one year, 95 percent of individuals assigned the lowest score (F.I.T. = 3) had died, whereas 96 percent of those assigned the highest scores (F.I.T. = 7-9) remained alive. The strongest predictors were the "I" and "T" parameters. CONCLUSION: This approach to assessment of candidates for bone marrow transplantation may identify individuals who require added, supportive measures, both medical and psychological. Furthermore, the results suggest possible leads in the search for how psychological factors might influence the physiologic response to a toxic stress. PMID- 10376232 TI - An educational intervention using the Agency for Health Care Policy and Research Depression Guidelines among internal medicine residents. AB - OBJECTIVE: To determine if a brief educational intervention utilizing the Agency for Health Care Policy and Research (AHCPR) Depression Guidelines would effect improved recognition of depressed patients, as well as improved attitudes and knowledge, among Internal Medicine housestaff. METHOD: This was a randomized trial of an educational intervention for Internal Medicine residents. All patients attending the resident clinics were screened using the Center for Epidemiologic Studies Depression Scale. Persons scoring greater than 16 constituted the prospective cohort. Three hundred eighty-four patients were screened for entry into the study. Of 160 persons meeting the entry criteria, follow up was available on seventy-two (60%). Residents were randomly assigned to receive the educational intervention which consisted of three sessions where the residents received copies and instructions about the AHCPR depression guidelines and the use of a case-finding instrument for depression. RESULTS: Non-recognized patients had milder symptoms of depression than did recognized patients. The presence of depressive symptoms was strongly related to measures of health status. Only seven of the seventy-two patients were identified as depressed and this was distributed equally between the two groups of residents. Intervention residents showed sustained improvement regarding depression criteria and the use of screening instruments at six months. CONCLUSIONS: A brief educational intervention effected changes in resident attitudes and knowledge regarding the care of depressed patients. Residents recognized patients with greater depressive symptoms than those with milder symptoms. PMID- 10376231 TI - The effects of major depression and phobia on stage at diagnosis of breast cancer. AB - OBJECTIVE: To examine the longitudinal effects of major depression and phobia on stage at diagnosis of subsequent breast cancer. METHOD: Data from the New Haven Epidemiologic Catchment Area (ECA) study were linked to the Connecticut Tumor Registry (CTR). The sample comprised of seventy-two women with a first primary breast cancer diagnosed sometime after their baseline ECA study interview. In the ECA study, lifetime psychiatric history was assessed using the Diagnostic Interview Schedule based on DSM-III criteria. Stage at diagnosis of breast cancer was taken from CTR records and dichotomized into early stage (in situ and localized tumors) versus late stage (regional and distant tumors). RESULTS: A positive history of major depression was associated with an increased likelihood of late-stage diagnosis of breast cancer (odds ratio [OR] = 9.81, p = 0.039), whereas a positive history of phobic disorders was associated with a decreased likelihood of late-stage diagnosis (OR = 0.01, p = 0.021), controlling for sociodemographic characteristics of the sample. CONCLUSIONS: These analyses revealed a longitudinal association between reported lifetime psychiatric history and stage at diagnosis of subsequent breast cancer. Phobia may motivate women to adhere to breast cancer screening recommendations and to report suspicious symptoms to a physician without delay. Major depression, on the other hand, was identified as an important predictor of late-stage diagnosis; proper recognition and management of depression in the primary care setting may have important implications for breast cancer detection and survival. PMID- 10376233 TI - Personality disorder traits: prevalence and effects on health status in primary care patients. AB - OBJECTIVE: Previous work in an academic setting has found that scoring in the higher ranges for selected personality disorders on an objective assessment tool was associated with increases in psychiatric co-morbidities, decreased satisfaction with health care, and diminished health related functional status. This study examines how often patients in primary care practices exhibit traits consistent with these selected disorders and what impact this has on their health related functional status and use of health care resources. METHODS: Thirteen family practices agreed to distribute questionnaires to 50 consecutive patients in the spring of 1997. Questionnaires contained instruments that assess risk for personality disorders, health related functional status, health resource use, demographics, and depression. The relationships between four specific personality disorders (borderline, dependent, schizoid and schizotypal) and other assessed variables were explored. RESULTS: Of the 250 patients returning completed survey instruments, 80 (32%) scored in the high range for traits consistent with one of the four target personality disorders. Patients in the high risk group also were noted to have more outpatient, emergency, and inpatient visits in the previous six months. Those in the high risk group also had significantly lower scores on seven of eight measures of health related functional status. CONCLUSIONS: Patients who have several traits for borderline, dependent, schizoid, and schizotypal personality disorders are common in primary care practices. These patients utilize services at higher rates than others and are more likely to screen in the positive range for depressive symptoms and have overall lower health related functional status. PMID- 10376234 TI - Patient-disease characteristics and coping strategies predict hospitalization in emergency psychiatry. AB - OBJECTIVE: The goal of this study was to analyze how far patient-disease characteristics (sociodemographic variables, previous psychiatric treatment, way of referral, the patient's current diagnosis), and the patient's coping strategies are connected with the consecutive disposition for inpatient or outpatient treatment. METHODS: Data from a one-year intake of the psychiatric emergency service at a University Hospital (N = 1439) were monitored and analyzed with regard to the decision on treatment. Four hundred eighty-one patients were hospitalized and 530 were assigned to outpatient treatment. Two subsamples of twenty-eight patients from each group filled out the Bernese Coping Modes questionnaire before the decision with regard to the treatment disposition was taken. RESULTS: The patient's psychiatric history, the way of referral as well as the current axis I diagnosis made a significant contribution to the treatment decision. Overall, patient-disease characteristics allowed for correct classification of 69.3 percent of cases. However, coping was a comparable predictor of hospitalization. CONCLUSIONS: It is argued that the search for patient-disease characteristics in the psychiatric emergency room should be complemented by a more extensive monitoring of the patients' way of coping with their current crisis. PMID- 10376235 TI - Commentary: the use of placebos in psychiatric research. PMID- 10376236 TI - Clonazepam treatment of panic disorder in patients with recurrent chest pain and normal coronary arteries. AB - OBJECTIVE: To examine the efficacy of clonazepam in chest pain patients with panic disorder and normal coronary arteries. METHOD: We conducted a placebo controlled, double blind, flexible dose (1-4 mg/d), six-week trial of clonazepam. All subjects (N = 27) had current panic disorder and a negative coronary angiogram or thallium exercise tolerance test within the previous year. RESULTS: Analyses show modest improvements in the clonazepam and placebo groups over the first four weeks in both primary outcome measures. Eight of twelve (67%) clonazepam treated patients responded with reduction of panic attacks by week four to zero per week or half of initial frequency, while seven of fifteen (47%) placebo treated patients responded (not significant). When response was measured by 50 percent reduction in Hamilton Anxiety total score, however, seven of twelve (58%) clonazepam treated patients responded, while two of fifteen (14%) placebo treated patients responded, (p = .038) by Fisher's exact test. Within-subject improvements over the first four weeks were not significantly greater for the clonazepam group than for the placebo group on either outcome measure. CONCLUSIONS: These results show a generally good outcome in chest pain patients with panic disorder, and they provide suggestive evidence for the efficacy of clonazepam compared to placebo. This study points to the need for larger, well funded treatment studies of chest pain patients with panic disorder. PMID- 10376237 TI - Treatment of panic disorder in older adults: a pilot study comparison of alprazolam, imipramine, and placebo. AB - OBJECTIVE: Several studies have documented that a variety of pharmacological compounds are quite effective in controlling acute symptomatology of panic disorder in the general population. However, there is a paucity of such studies in the management of panic disorder in older adults (ages 55 and above). The purpose of this study was to gather pilot data in older patients with panic disorder to begin to assess the efficacy of two commonly-used antipanic medications, imipramine and alprazolam. METHOD: Twenty-five (n = 25 (23 females; 2 males); 18 completers, 7 dropouts) older panic disorder (DSM-III-R) patients (age range = 55-73; mean = 61.24) were studied in an eight-week randomized, parallel-groups, double-blind, placebo-controlled, flexible dose design. Outcome was assessed weekly by global change ratings (Hamilton Anxiety and Depression Scales; Physicians' Global Impression ratings) and panic diaries. Because of small sample size, we present data descriptively. RESULTS: Subjects in active medication groups evidenced reductions in panic attacks and in level of overall anxiety and depression. Therapeutic dosages were approximately half those commonly used in younger panic disorder patients. CONCLUSION: Our data suggest the comparable efficacy of alprazolam and imipramine in the short-term treatment of older adults with panic disorder. There is clearly the need for a larger scale placebo-controlled study, preferably comparing imipramine and/or alprazolam with one of the SSRIs, to substantiate our findings. PMID- 10376238 TI - Effect of feeding before, during and after milking on milk production and the hormones oxytocin, prolactin, gastrin and somatostatin. AB - Feeding during milking has been shown to influence milk production, milk flow and milking time as well as the secretion of the pituitary hormones oxytocin and prolactin, and the gastrointestinal hormone somatostatin. However, it is not known whether feeding before or after milking has any effect. The aim of the present study was to investigate how the timing of feeding relative to milking influences milk production and flow, milking time and hormone secretion. The trial was carried out over 9 weeks with 24 cows at varying stages of lactation. Each treatment period lasted for 3 weeks, including one registration week. The cows were fed ad lib. and were exposed to three treatments: feeding 1.5 h before milking (FBM), feeding at exactly the same time as milking (FDM) and feeding 1.5 h after milking (FAM). The most marked treatment effect was observed during morning milking. FDM resulted in higher milk production and higher yields of protein and lactose. FAM produced a lower fat yield and a lower fat content compared with FDM, and a lower lactose content than either FBM and FDM. Milking time was longer when cows were fed during milking, but no significant effects on milk flow were found. The amount of milk collected during the first 2 min of milking was lower when cows were fed after milking. Milking-related oxytocin and somatostatin secretion was lower in FAM than in FDM. The level of prolactin was lower when cows were fed before or after than during milking. More studies are needed to elucidate whether there is a long-term effect on milk production related to the discussed milking routines. PMID- 10376240 TI - Effects of calcium soaps and rumen undegradable protein on the milk production and composition of dairy ewes. AB - Forty-eight Manchega dairy ewes were used during a complete lactation in a 2 x 2 factorial design to determine the effects of supplementing diets with fat (calcium soaps of palm oil fatty acids, CSFA) and rumen undegradable protein (RUP) on milk production and composition. Factors tested were amounts of CSFA (0 or 200 g/kg) and RUP (300 or 450 g/kg crude protein) in the concentrate. RUP was altered by adding a mixture of maize gluten meal and blood meal. Lactation was divided into one nursing period (period 1, weeks 1-4), and three milking periods (periods 2-4, weeks 5-8, 9-14 and 15-21). Concentrates were given at 0.8 kg/d during periods 1 and 2, and at 0.6 kg/d in periods 3 and 4. Ewes grazed rotationally in an Italian rye-grass pasture and received a daily supplement of 0.8 kg vetch-oat hay during period 1, and 0.3 kg lucerne hay during periods 2-4. For the whole lactation, supplemental fat markedly increased milk fat content (+23%) and yield (+16%), and decreased milk protein content (-9%). The positive effect of feeding CSFA on milk fat content was more evident at the beginning of lactation; however, its negative effect on milk protein was more pronounced in late lactation. Supplementary RUP had little effect, increasing milk protein content only in period 3, when the crude protein content of pasture was lower. Milk yield and lamb growth were not affected by dietary treatments. The results indicated that CSFA can be useful for increasing the milk fat content of dairy ewes at pasture, which may help farmers to produce milk reaching the minimum requirements of fat content for the cheese industry. PMID- 10376239 TI - Effects of stocking density and concentrate supplementation of grazing dairy cows on milk production, composition and processing characteristics. AB - The effects on milk composition and processing characteristics of varying grass supply by changing stocking density and of offering a concentrate supplement were investigated. The experiment was conducted over 28 weeks of the lactation (April October) using 48 spring-calved Friesian-Holstein cows. Three herds each of 16 cows were offered a restricted grass supply, a standard grass supply and a standard grass supply with a supplement of 3 kg concentrate/d. Treatment groups were grazed separately with a residence time of 3 d/paddock. Milk production, composition and processing characteristics such as renneting properties, ethanol stability and plasmin activity were measured weekly. Increasing stocking density above the standard system resulted in significant reductions in milk fat and protein yields, the concentrations of total protein, casein and whey proteins, and a deterioration in most processing characteristics. Imposing concentrate supplementation on the standard system increased total protein, casein and whey protein concentrations but generally did not improve processing characteristics except for ethanol stability. These results suggest that the standard grass supply in a rotational grazing paddock system can support efficient production of quality milk, and concentrate supplementation will not improve processing characteristics when an adequate supply of good quality herbage is available. PMID- 10376241 TI - Effects of dietary protein supply on caseins, whey proteins, proteolysis and renneting properties in milk from cows grazing clover or N fertilized grass. AB - The objective of this work was to examine whether variation in the amino acid supply to cows could be a reason for the reduced casein content and poorer renneting properties of milk that often occur in late summer, or whether these effects are related to proteolysis in the raw milk. In a 2 x 2 x 2 factorial design, we investigated the effects of sward (clover v. rye-grass) and supplementary feed with a high or low level of rumen-soluble N or of rumen undegradable protein on milk protein composition during the grazing season. A total of 32 Danish Holstein cows were included in the experiment. Milk protein and casein contents and the ratios casein N:total N and casein:true protein were at a minimum in late summer, whereas the contents of urea, non-protein N and whey protein were higher during this period. These seasonal effects were unrelated to either the type of supplementary feed or the type of sward; neither were they clearly related to proteolysis, although casein: true protein was related to the proteose peptone content. The results indicated that whey proteins other than alpha-lactalbumin or beta-lactoglobulin accounted for the higher proportion or concentration of whey protein in late summer. Based on a principal component analysis including variables such as citric acid, lactose and non-protein N, we suggest that the cows' energy supply during this period may be a critical factor in determining the milk protein composition, although our results were not conclusive. There was an interaction between the supplement of rumen undegradable protein and type of sward. When clover was grazed, a high supplement increased the concentrations of protein and casein in milk and the kappa-casein: total casein ratio. When rye-grass was grazed, the opposite response was found, and overall milk protein yield was not affected. The very low N content of clover in early summer reduced milk protein and casein protein during this period. PMID- 10376242 TI - Changes in plasma and milk concentrations of glucose and IGF-1 in response to exogenous growth hormone in lactating goats. AB - Exogenous growth hormone was administered subcutaneously to five lactating goats during the post-peak period of lactation. Milk yields increased significantly by approximately 20% in response to growth hormone. Blood and milk samples were taken in the periods before, during and after growth hormone treatment. The concentrations of glucose in milk increased significantly by approximately 50% in the period following growth hormone treatment at a time corresponding to the increase in milk yield. There was a transient increase in plasma glucose concentrations immediately following growth hormone treatment before either milk glucose concentrations or milk yields were increased. Both free and total IGF-1 concentrations in plasma increased slowly following growth hormone treatment. The increase in plasma IGF-1 corresponded to the increase in milk yields and milk glucose concentrations. Concentrations of IGF-1 in milk increased more rapidly than those in plasma, rising by approximately 150% following growth hormone treatment, and were starting to decline by the time that milk yield and milk glucose concentrations were at their maximum. As milk glucose concentrations have been shown previously to reflect changes in the intracellular concentration of glucose, the results indicate that part of the mechanism by which growth hormone stimulates milk production is by increasing the intracellular availability of glucose for lactose synthesis. The results also suggest that changes occur in the concentrations of IGF-1 in the environment of the mammary gland before changes are observed in the general circulation, and that these are reflected in the changed concentrations in milk. PMID- 10376243 TI - Process steps for the preparation of purified fractions of alpha-lactalbumin and beta-lactoglobulin from whey protein concentrates. AB - Fractions enriched with alpha-lactalbumin (alpha-la) and beta-lactoglobulin (beta lg) were produced by a process comprising the following successive steps: clarification-defatting of whey protein concentrate, precipitation of alpha lactalbumin, separation of soluble beta-lactoglobulin, washing the precipitate, solubilization of the precipitate, concentration and purification of alpha-la. The present study evaluated the performance of the process, firstly on a laboratory scale with acid whey and then on a pilot scale with Gouda cheese whey. In both cases soluble beta-lg was separated from the precipitate using diafiltration or microfiltration and the purities of alpha-la and beta-lg were in the range 52-83 and 85-94% respectively. The purity of the beta-lg fraction was higher using acid whey, which does not contain caseinomacropeptide, than using sweet whey. With the pilot scale plant, the recoveries (6% for alpha-la; 51% for beta-lg) were disappointing, but ways of improving each step in the process are discussed. PMID- 10376244 TI - Deterioration of protein fraction by Maillard reaction in dietetic milks. AB - The development of the Maillard reaction in pasteurized, UHT and in-bottle sterilized dietetic milks was studied. In these products damage caused by heat treatments could increase as a result either of the addition of various ingredients or of manufacturing processes that alter their content of reducing carbohydrates. Protein damage was evaluated by measuring furosine by reversed phase ion-paired HPLC. The levels of furosine detected made it possible to assess the amounts of biologically unavailable lysine. In all milks analysed blocked lysine values were < 340-350 mg/g total lysine, the level at which lysine becomes the limiting amino acid in milk. Pasteurized dietetic milks had levels of blocked lysine similar to that in ordinary pasteurized cows' milk. In some UHT and in bottle sterilized dietetic milks their different composition resulted in an increase in the blocked lysine content. In some in-bottle sterilized milks, protein damage greatly reduces the beneficial effects of milk as a dietary supplement. Lactose-free milks, which are more susceptible to protein deterioration because of their higher content of reducing carbohydrates, were also analysed after storage at 20 degrees C and at < or = 4 degrees C. At the end of their recommended storage times, they contained limited amounts of blocked lysine only if they had been stored at < or = 4 degrees C. PMID- 10376245 TI - Purification and characterization of a lysine-p-nitroanilide hydrolase, a broad specificity aminopeptidase, from the cytoplasm of Lactococcus lactis subsp. cremoris AM2. AB - A hydrolase activity that cleaves lysyl-p-nitroanilide (Lys-pNA) has been purified from the cytoplasm of Lactococcus lactis subsp. cremoris AM2 by chromatography on DE52, DEAE Affi-Gel Blue Gel, Hydroxyapatite Bio-Gel HTP and Phenyl Sepharose. The purified aminopeptidase was found to have a native M(r) of 50,000-55,000 by gel filtration chromatography and by FPLC gel filtration on Superose 12 and to be composed of a single polypeptide chain following SDS-PAGE. Enzyme activity was almost completely inhibited by EDTA, amastatin, puromycin and bestatin, while the sulphydryl-reactive agents p-chloromercuribenzoate and iodoacetamide were inhibitory. The enzyme was found to be very unstable during the purification procedures at 4 degrees C and its stability was greatly improved when 10 ml glycerol/l and 2 mM-dithiothreitol were included in the purification buffers. The purified enzyme was found to hydrolyse a wide range of dipeptides, tripeptides and longer peptides provided that proline was not present in the penultimate position from the N-terminus or that a pyroglutamyl residue was not present at the N-terminus. While neither Asp-pNA nor Pro-pNA was hydrolysed by the purified enzyme, the release of N-terminal acidic residues from peptides was observed in addition to the release of N-terminal proline from Pro-Leu-Gly-NH2, Pro-Leu-Gly-Gly and Pro-His-Pro-Phe-His-Leu-Phe-Val-Tyr. This ability of Lys-pNA hydrolase to release N-terminal proline residues was employed in concert with a purified aminopeptidase P preparation to release alternate N-terminal amino acids from Tyr-Pro-Phe-Pro-Gly. The complementary action of these enzymes represents an alternative mechanism to that of post-proline dipeptidyl aminopeptidase for metabolism of proline-containing peptides. PMID- 10376246 TI - 15N enrichment of casein amino acids in the milk from goats given a single intravenous dose of L-[15N]leucine. PMID- 10376247 TI - Expression of bovine beta-lactoglobulin in the milk of transgenic mice. PMID- 10376248 TI - Multiple forms of lactadherin (breast antigen BA46) and butyrophilin are secreted into human milk as major components of milk fat globule membrane. PMID- 10376249 TI - DNA probe for Lactobacillus delbrueckii subsp. lactis. PMID- 10376250 TI - Single-step method for rapid detection of Brucella spp. in soft cheese by gene specific polymerase chain reaction. PMID- 10376251 TI - Characterization of kefir grains grown in cows' milk and in soya milk. PMID- 10376253 TI - Peripheral paraphrasing the glaucoma lexicon. PMID- 10376252 TI - Sugar formulation effect on available lysine content of dulce de leche. PMID- 10376254 TI - Effect of pupillary dilation on retinal nerve fiber layer thickness as measured by scanning laser polarimetry in eyes with and without cataract. AB - PURPOSE: This study was conducted to evaluate the effect of pupillary dilation on retinal nerve fiber layer (RNFL) thickness as measured by scanning laser polarimetry (SLP) in cataractous and noncataractous eyes. METHODS: The study included 31 eyes of 31 consecutive patients (mean age, 62.5 +/- 14.0 years; range, 30-76 years). Eyes with refractive error exceeding 5.0 D sphere or 2.0 D cylinder, nonlenticular media opacity, cup-to-disc ratio > 0.9, corneal disease, ocular inflammation, or previous intraocular surgery were excluded. A standard reticule was used to measure pupillary diameter. Cataract grade was evaluated by a single observer using the Lens Opacities Classification System (LOCS III). RNFL thickness measurements were obtained by means of SLP before and after pupillary dilation. RESULTS: Of the patients, 10 had clear lenses and 21 had variable degrees of lenticular opacification. In four eyes, imaging could not be performed because of dense cataracts. Mean pupillary diameters before and after dilation were 2.5 +/- 0.7 mm and 7.3 +/- 1.1 mm, respectively. There were no significant differences in global RNFL thickness before and after dilation in noncataractous and cataractous eyes. Among cataractous eyes in which imaging was possible, there was no correlation between difference in RNFL thickness before and after dilation and nuclear opalescence, nuclear color, and cortical and posterior subcapsular grading of the LOCS III score. Six of 27 eyes (22.2%) had a change of more than 10% in RNFL thickness after pupillary dilation. CONCLUSION: Although pharmacologic mydriasis does not statistically alter RNFL thickness as measured by SLP, approximately one fifth of such eyes will have a change of more than 10% in retardation. Uniformity in pupil size is recommended when longitudinally evaluating RNFL measurements. PMID- 10376255 TI - Diversity of response of optic nerve head circulation to timolol maleate in gel forming solution. AB - PURPOSE: This randomized, double-masked, placebo-controlled, two-period crossover study was conducted to investigate the effects of 0.5% timolol maleate in gel forming solution on intraocular pressure (IOP) and blood circulation in the optic nerve head in patients with untreated ocular hypertension. METHODS: The effects of 0.5% timolol in gel-forming solution on IOP and optic nerve head capillary blood speed were studied in 12 patients with untreated ocular hypertension. Optic nerve capillary blood speed was measured using the laser Doppler technique before and at the end of each treatment period. RESULTS: In each patient, IOP decreased after treatment with timolol (mean decrease 16.8% versus placebo). Systemic blood pressure and pulse rate did not differ significantly after treatment with topical timolol from values after placebo. The mean change from baseline in Doppler broadening was 10.6% greater after treatment with timolol than after placebo. There was no significant change in mean Doppler broadening from baseline after treatment with either timolol or placebo. However, optic nerve head capillary blood speed increased in six patients, and was within the range of placebo response in six patients after treatment with timolol. Spearman correlation analysis of the baseline with Doppler broadening measurements after treatment showed a correlation for placebo but not for timolol. The percent change in Doppler broadening after timolol treatment was correlated with iris color. CONCLUSION: These results indicate that administration of timolol for 4 weeks reduces IOP in patients with ocular hypertension and generally does not change the blood circulation in the optic nerve head. Individual patients, however, showed variable changes in optic nerve head circulation after topical administration of timolol. Although the sample size was small, these changes in optic nerve head circulation were correlated with iris color. PMID- 10376256 TI - Correlation of functional and structural measurements in eyes suspected of having glaucoma. AB - PURPOSE: This study was conducted to determine the correlation between structural changes in the retinal nerve fiber layer (RNFL) and functional loss detected on short-wavelength automated perimetry (SWAP) in a population of patients with suspected glaucoma. METHODS: With a selection criteria of intraocular pressure (IOP) more than 21 mmHg and normal results of conventional automated perimetry, 49 eyes of 49 patients with ocular hypertension were enrolled in the study. The SWAP was performed with a modified Humphrey field analyzer, and visual field indexes (mean deviation [MD], corrected pattern standard deviation [CPSD]) were calculated. Semiquantitative RNFL scores were given separately to diffuse and localized defects of the RNFL. RESULTS: The MD increased significantly with higher diffuse and total RNFL scores, with good correlation coefficients. A weak correlation was found between CPSD and diffuse, total, and localized RNFL scores. CONCLUSION: Diffuse RNFL loss are associated with abnormalities in visual field indexes (MD), whereas focal structural damage showed no correlation with visual field loss. PMID- 10376257 TI - Audible pops during cyclodiode procedures. AB - PURPOSE: This study was conducted to determine the incidence of intraocular uveal microexplosions ("pops") during contact diode laser transscleral cyclophotocoagulation (cyclodiode) and to analyze the influence of pop occurrence on results and postoperative complications. METHODS: Cyclodiode treatment (1.5-2 W x 2 seconds over 270 degrees) was performed in 43 consecutive patients (43 eyes) with uncontrolled glaucoma who had not undergone previous ciliary ablation. Mean duration of follow-up evaluation was 12.7 months (range, 9-18 months). These eyes included 31 seeing eyes, in which intraocular pressure (IOP) reduction was indicated to preserve visual acuity (therapeutic group) and 12 blind eyes, in which IOP reduction was advisable to relieve pain (palliative group). Success was defined as a final IOP > or = 5 mmHg and < or = 21 mmHg in seeing eyes, and as the resolution of pain in blind eyes. Potential factors evaluated for intraoperative pop occurrence included patient age, gender, iris color, glaucoma diagnosis, preoperative IOP, and number of previous surgical procedures used to treat glaucoma. RESULTS: In the group receiving therapeutic treatment, mean +/- standard deviation (SD) IOP was 41.8 +/- 12.7 mmHg before surgery and 19.2 +/- 8.3 mmHg after surgery; the success rate in this group was 83.9%. In the group receiving palliative treatment, mean IOP was 55.4 +/- 13.7 mmHg before surgery and 21.1 +/- 13.4 mmHg after surgery; success rate was 83.3% in this group. Intraoperative microdisruptions occurred in 48.8% of the cases during the first laser application; 67.5% of these occurred in the superior half of the eye. No significant difference in rate of intraoperative pop was observed between patients after one cyclodiode session and those eyes that underwent additional sessions. Mean baseline IOP was significantly higher in patients with intraoperative occurrence of pops. An audible pop was more common in the group undergoing palliative treatment. Intraoperative occurrence of pops was associated with a greater severity of postoperative iridocyclitis. All patients with postoperative hyphema also had pops during surgery. No significant difference in the success rate was found between patients in whom intraoperative pops did and did not occur. CONCLUSION: Choroidal vaporization is significantly more common in patients with higher baseline IOP. Occurrence of pops also is associated with more severe postoperative inflammation and with a greater risk of postoperative hyphema. PMID- 10376258 TI - Mitomycin-C in combined or two-stage procedure trabeculectomy followed by penetrating keratoplasty. AB - PURPOSE: To evaluate the efficacy and safety of application of mitomycin-C (MMC) in combined and separate trabeculectomy and penetrating keratoplasty for the treatment of coexisting corneal disease and glaucoma. METHODS: A retrospective evaluation of 11 eyes of 10 patients was conducted. A combined trabeculectomy with MMC and penetrating keratoplasty procedure was performed in eight eyes (group 1), and keratoplasty was performed after a previous trabeculectomy with MMC in three eyes (group 2). RESULTS: In group 1, seven of the eight eyes had controlled intraocular pressure (IOP) and clear corneal graft at the end of the follow-up period (range, 5-60 months; mean duration, 16.7 months). In group 2, all three eyes had controlled IOP at the end of the follow-up period (range, 4-30 months; mean duration, 14 months); two of these patients had clear corneal grafts, and graft failure occurred in the remaining patient. Complications included transient flat anterior chamber and corneal epithelial defects, each of which occurred in a single eye in group 1, and progressive cataract, which occurred in a single eye in group 2. CONCLUSION: Mitomycin-C was found to be safe and efficient in the present series, controlling IOP in 10 of 11 eyes (91%) with coexisting corneal disease and glaucoma. The transplanted corneas remained clear in 9 of 11 eyes (82%). Complications related to MMC included a reversible epithelial defect that occurred in one eye. PMID- 10376259 TI - Trabeculectomy using a sutureless scleral tunnel technique: a preliminary study. AB - PURPOSE: This study was conducted to evaluate prospectively the surgical outcome in terms of intraocular pressure (IOP) control and safety of trabeculectomy using a sutureless scleral tunnel technique in patients with uncomplicated primary open angle glaucoma (POAG). METHODS: Patients with POAG whose IOP was more than 21 mmHg with administration of maximally tolerated medications were recruited for this study. All patients underwent the sutureless scleral tunnel trabeculectomy under retrobulbar anesthesia. Intraoperative complications and postoperative visual acuity, IOP, bleb status, and complications were evaluated for a follow-up period of 1 year. RESULTS: Surgery was performed in 18 patients (20 eyes); 17 of the patients (19 eyes) completed the study (one patient was excluded because of defaulted follow-up). Mean IOP was 26.6 +/- 3.6 mmHg before surgery and 15.3 +/- 3.0 mmHg 1 year after surgery. No intraoperative complications were encountered. Mild hyphema (10.5%) and shallow anterior chamber (15.8%) were noted in the immediate postoperative period, but both were managed successfully with conservative treatment. The absolute success rate was 79.0%. CONCLUSION: The preliminary results of this study are encouraging. It appears that sutureless scleral tunnel trabeculectomy is a safe and effective drainage procedure for treating uncomplicated POAG. A larger-scale study with a longer follow-up period seems warranted. PMID- 10376260 TI - Mitomycin C in higher risk trabeculectomy: a prospective comparison of 0.2- to 0.4-mg/cc doses. AB - PURPOSE: This randomized, masked, prospective study was conducted to compare the outcome of filtering surgery using doses of 0.2 mg/cc or 0.4 mg/cc of mitomycin C (MMC) in eyes that were at higher risk from previous conjunctival incisional surgery. METHODS: Eyes of 50 consecutive patients with primary open-angle, pseudoexfoliation, or pigmentary glaucoma requiring trabeculectomy who had previously undergone either limbal cataract surgery or trabeculectomy were enrolled. Patients received an intraoperative dose of either 0.2 or 0.4 mg/cc MMC for 2 minutes (n = 25 in each study group). Intraocular pressure (IOP), logMAR visual acuity, and complications were monitored at regular intervals for 1 year. Unpaired student t tests were used to compare percent decrease in IOP in both study groups at each interval. RESULTS: The percent decrease in IOP was not significantly different between groups at 1 day, 1 week, 1 month, 3 months, 6 months, or 1 year after surgery. LogMAR visual acuity was not significantly different between groups at 1 month, 6 months, or 1 year. Treatment failure occurred in seven patients in the 0.2 mg/cc group (28%) and seven patients in the 0.4 mg/cc group (29.2%). Postoperative hypotony, choroidal effusions and hemorrhages, and wound leaks occurred in both groups, but occurred more often in the group receiving 0.4 mg/cc MMC. CONCLUSION: Filtering surgery performed on higher risk eyes was as effective using a lower dose (0.2 mg/cc) of MMC as that using a higher dose (0.4 mg/cc). Incidence of complications and treatment failures was slightly higher in the group receiving high-dose MMC. PMID- 10376262 TI - A randomized trial of the effect of midazolam on intraocular pressure. AB - PURPOSE: The effect of midazolam on intraocular pressure (IOP) in adults was studied as an initial step in determining whether it can be used as a preoperative anxiolytic or sedative agent in children with glaucoma who are undergoing examination for IOP measurements. METHODS: This study followed a prospective, placebo controlled, randomized, double masked design. Fifty-five participants were enrolled after informed consent was obtained. Each enrolled patient underwent a brief history and eye examination. Measurements of IOP were taken at baseline and 5, 10, and 15 minutes after intravenous administration of 1 mg midazolam or placebo. IOP was the primary outcome measured. RESULTS: There was no difference in IOP fluctuation from baseline between patients who received midazolam and those who received placebo. CONCLUSION: Early results indicate that because midazolam does not lower IOP, it may be a useful adjunct to ketamine anesthesia in children with glaucoma undergoing ophthalmologic examination under anesthesia. However, studies of midazolam must be conducted in children and patients with glaucoma before its use in these populations can be recommended. PMID- 10376261 TI - Latanoprost in glaucoma associated with Sturge-Weber syndrome: benefits and side effects. AB - PURPOSE: This study was conducted to evaluate the ocular hypotensive efficacy, safety, and side effects of latanoprost 0.005% administered as adjunctive therapy in patients with Sturge-Weber syndrome (SWS) and glaucoma. METHODS: Commercially available latanoprost 0.005% was added as a single drop once daily to other antiglaucoma medications. Intraocular pressure (IOP) was measured at 1, 3, and 6 months of treatment. A successful response was defined as a reduction of at least 20% in IOP at the final follow-up evaluation without additional medical or surgical therapy and no adverse events related to latanoprost. RESULTS: 18 eyes of 18 patients with SWS and glaucoma were enrolled from 9 clinical centers. Mean baseline IOP was 28.4 +/- 7.1 mmHg (range, 17-42 mmHg). Using Kaplan-Meier analysis, a successful response to latanoprost was observed in 3 of 18 (16.7%) patients at the 6-month interval. Seven (38.9%) patients required surgery; three (16.7%) patients required additional medical therapy, seven (38.9%) patients had no change in therapy. One (5.6%) patient discontinued latanoprost treatment because of intolerable conjunctival hyperemia. Two successfully treated patients had significantly greater episcleral vessel engorgement after initiation of latanoprost therapy. CONCLUSION: Patients with SWS and glaucoma respond poorly to adjunctive latanoprost therapy and often require additional medical or surgical intervention. Increased episcleral vascular engorgement might result in greater operative risks should filtration surgery become necessary in these patients. PMID- 10376263 TI - Recurrent angle-closure glaucoma. PMID- 10376264 TI - Vascular dysregulation: a principal risk factor for glaucomatous damage? AB - Both intraocular pressure (IOP) and vascular factors appear to play an important role in the pathogenesis of glaucomatous optic neuropathy (GON). Arteriosclerosis and its risk factors are of minor importance, whereas vasospastic syndrome clearly is associated with GON. A vascular endotheliopathy seems to be involved in the diathetic hyperresponsiveness to stimuli, such as coldness or emotional stress. This in turn leads to a compromised autoregulation, and thereby renders the eye more sensitive to IOP or to a decrease in blood pressure. A variation in ocular perfusion may lead to an increase in free oxygen radicals. This may finally lead to apoptosis. PMID- 10376265 TI - Change in optic disc topography associated with diurnal variation in intraocular pressure. AB - PURPOSE: A case demonstrating significant changes in optic disc topography from variation in intraocular pressure (IOP) is demonstrated. METHODS: Within a single 24-hour period, confocal scanning laser ophthalmoscopic images were obtained in the left eye of a patient during an IOP peak (60 mmHg) and during an IOP trough (18 mmHg). RESULTS: During an IOP trough, paired t tests showed significant reduction in cup area, cup volume, mean cup depth, and maximum cup depth, and a significant increase in rim area and cup-to-disc (CD) ratio. CONCLUSION: Under certain conditions, optic disc topography may be affected by diurnal variations in IOP. PMID- 10376266 TI - Putative role of placental corticotropin-releasing factor in the mechanisms of human parturition. AB - Corticotropin-releasing factor is a 41-amino-acid neuropeptide synthesized in the paraventricular nucleus of the hypothalamus and released in response to stress. Its major role is the regulation of the hypothalamus-pituitary-adrenal axis by stimulation of ACTH release from the anterior pituitary gland. In addition, corticotropin-releasing factor modulates behavioral, vascular, and immune responses to stress. Corticotropin-releasing factor was first detected in the extracts of human placentas obtained at full term from spontaneous deliveries. Placental corticotropin-releasing factor content and messenger RNA expression progressively increase during normal pregnancy, and corticotropin-releasing factor levels in maternal plasma have a similar time course. The addition of corticotropin-releasing factor to primary trophoblast cell cultures stimulates ACTH secretion in a dose-dependent manner, and its action is mediated by cyclic adenosine monophosphate as second messenger. In addition, corticotropin-releasing factor is a potent local regulator of myometrial contractility and of membrane prostaglandin release. The effects of corticotropin-releasing factor in these various tissues are mediated by specific receptors. Placental corticotropin releasing factor is also secreted into the fetal circulation and the stimulation of fetal pituitary ACTH and fetal adrenal gland dehydroepiandrosterone sulfate release in vitro has been shown. Recently, urocortin, a new peptide related to corticotropin-releasing factor, has been found in human placenta. Corticotropin releasing factor and urocortin share some of their biologic effects, acting on the same receptors. A large-molecular-weight corticotropin-releasing factor binding protein modulates the activity of both these peptides. Plasma corticotropin-releasing factor levels are low in nonpregnant women and become higher during the second trimester of pregnancy, rising steadily until about 35 weeks, and then increasing more rapidly until term. Vaginal delivery is a condition associated with the highest values of maternal corticotropin-releasing factor levels. Corticotropin-releasing factor is also measurable in fetal plasma (20-fold lower than in maternal circulation) and in amniotic fluid. Increased maternal plasma corticotropin-releasing factor levels characterize some gestational diseases. Women with chronic hypertension and preeclampsia have high corticotropin-releasing factor levels, and intrauterine growth retardation is associated with an activation of the hypothalamus-pituitary-adrenal axis, reflected by increased fetal plasma concentrations of ACTH, cortisol, and corticotropin-releasing factor. The role of corticotropin-releasing factor in preterm labor is uncertain, but midgestational plasma corticotropin-releasing factor levels may be higher in women delivering preterm. In these various pathologic states, maternal plasma corticotropin-releasing factor-binding protein levels undergo opposite changes, decreasing to very low levels. The endocrine paracrine corticotropin-releasing factor/corticotropin-releasing factor-binding protein pathways may play a major role in the mechanism of human parturition. PMID- 10376267 TI - Transient umbilical cord occlusion in late-gestation fetal sheep results in hippocampal damage but not in cerebral arteriovenous difference for nitrite, a stable end product of nitric oxide. AB - OBJECTIVE: To investigate the effect of total umbilical cord occlusion on cerebral arteriovenous difference for nitrite (a stable end product of nitric oxide) and neuronal outcome. METHOD: The cord was totally occluded for 10 minutes in 14 late-gestation (gestational age 113-120 days) chronically instrumented fetal sheep. Arterial (carotid artery) and venous (superior sagittal sinus) blood samples were taken at regular intervals for determination of acid-base status, glucose, lactate, and nitrite plasma levels. Three days after the occlusion period the fetal brain was perfusion fixed, and the parietal cortex, hippocampus, and cerebellum were scored for neuronal damage. RESULTS: Three fetuses died shortly after the occlusion period. Total umbilical cord occlusion resulted in a combined respiratory and metabolic acidosis as observed in carotid arterial blood gas samples (pH, 6.96 +/- 0.02; CaO2 [mmol/L], 0.43 +/- 0.9; PaCO2 [kPa], 13.46 +/- 0.38; base excess [mmol/L], -11.1 +/- 0.8; lactate [mmol/L], 10.57 +/- 0.95; bradycardia, 75 +/- 9 bpm; and hypotension, 29.85 +/- 3.00 mmHg) (n = 14, values are mean +/- standard error of the mean). Selective brain damage was observed in the hippocampus in 10 of the 11 surviving fetal sheep. No changes in arterial, venous, or cerebral arteriovenous difference for nitrite plasma levels were observed (n = 11). CONCLUSION: Total umbilical cord occlusion of 10 minutes in preterm fetal sheep results in hippocampal damage but not in changes of cerebral arteriovenous difference for nitrite plasma levels, a stable end product of nitric oxide. PMID- 10376268 TI - Prostaglandin endoperoxide synthase-2 abundance is increased in brain tissues of late-gestation fetal sheep in response to cerebral hypoperfusion. AB - OBJECTIVE: To determine the mechanism by which cerebral hypoperfusion enhances prostanoid secretion by fetal brain tissues. METHODS: Studies were performed on five intact and five carotid sinus-denervated sheep fetuses (124-136 days) exposed to 10 minutes of cerebral hypoperfusion. Plasma collected from lingual artery and sagittal sinus, and microdialysates collected from brain stem and hypothalamus were assayed for prostanoid production. Fetal hypothalamus, cerebral cortex, hippocampus, cerebellum, and brain stem were collected from intact animals and 30 minutes after cerebral hypoperfusion for the expression, activity, and distribution of prostaglandin endoperoxide synthase-1 (PGHS-1), PGHS-2, and thromboxane synthase. RESULTS: Thromboxane B2 increased significantly in sagittal sinus compared with arterial blood, but PGE2 did not change. Thromboxane B2 decreased in brain stem and hypothalamus microdialysates, and prostaglandin E2 increased in these regions. PGHS-2 immunoreactive protein levels in brain tissues increased in the cerebral hypoperfusion fetuses compared with those of the intact animals. By contrast, PGHS-1 and thromboxane synthase protein levels did not change between these two groups. Prostaglandin endoperoxide synthase activity in brain tissues decreased with the increased levels of immunoreactive PGHS-2. CONCLUSIONS: 1) Prostanoids are produced in response to cerebral hypoperfusion, 2) the increase in the production of prostanoid responses to cerebral hypoperfusion is associated with the decrease in activity of, and therefore, the "suicide" inactivation of PGHS, and 3) PGHS-2 is the predominant form of PGHS, whose synthesis is induced by cerebral hypoperfusion in the fetal brain. PMID- 10376269 TI - Increased neutrophil-endothelial adhesion induced by placental factors is mediated by platelet-activating factor in preeclampsia. AB - OBJECTIVE: Endothelial cell activation or dysfunction and neutrophil-endothelial cell adhesion have been suggested to be important in the pathophysiology of preeclampsia. However, the mechanisms that underlie the alteration of endothelial cell function in preeclampsia are unknown. Placenta from preeclamptic pregnancies produces mediators and autacoids, which may be released into the maternal circulation and modulate endothelial function. In this study, the effect of placental factor(s) on neutrophil-endothelial adhesion and the possible role of platelet-activating factor (PAF) in mediating the response have been examined. METHODS: Endothelial cells were isolated from human umbilical veins (HUVECs) from normal pregnancies. Confluent primary passage HUVECs were exposed to conditioned medium derived from normal and preeclamptic placental tissue cultures, with unconditioned medium as a control. Placental-conditioned medium was prepared by incubation of placental whole villous tissue in Dulbecco's Modified Eagle's Medium (DMEM) for 48 hours. Neutrophil-endothelial adhesion assays were performed to evaluate placental factors in mediating neutrophil-endothelial adhesion, and a PAF-3H scintillation proximity assay (SPA) system was used to determine endothelial PAF production. The PAF-receptor antagonist WEB 2086 was used to block placental factor-mediated increased neutrophil-endothelial adhesion induced by conditioned medium derived from preeclamptic placenta. RESULTS: Neutrophils were significantly more adherent to HUVECs treated with conditioned medium from preeclamptic placentas (28.44 +/- 2.47%) than to HUVECs treated with conditioned medium from normal placentas (18.95 +/- 1.57%) or with unconditioned medium (14.60 +/- 1.29%, P < .01). Also, HUVECs exposed to preeclamptic placental conditioned medium produced more PAF than the cells exposed to normal conditioned medium and unconditioned medium, 416.18 +/- 17.14 pg/1 x 10(7) cells versus 330.90 +/- 35.70 and 296.43 +/- 44.40 pg/1 x 10(7) cells, P < .05, respectively. The PAF receptor antagonist WEB 2086 completely blocked increased neutrophil endothelial adhesion induced by preeclamptic placental-conditioned medium (13.24 +/- 0.81% versus 31.31 +/- 4.75%, P < .01). CONCLUSION: In preeclampsia, the placenta releases one or more factors promoting neutrophil-endothelial adhesion. The increased neutrophil-endothelial adhesion thereby induced is a PAF-mediated event. It is suggested that if preeclamptic placentas release toxic factors into the maternal circulation in vivo, these factors may contribute to the altered vascular endothelial cell function in preeclampsia. PMID- 10376270 TI - Endothelial monocyte-activating polypeptide-2 is increased in pregnancy but is not further increased in preeclampsia. AB - OBJECTIVES: Endothelial monocyte-activating polypeptide-2 (EMAP-2) is a novel protein that demonstrates potent proinflammatory activity in vivo and in vitro. The purpose of this study was to investigate the expression of EMAP-2 in normal pregnancy and in pregnancies complicated with preeclampsia and to test the hypothesis that EMAP-2 is a causative agent of the endothelial activation of preeclampsia. METHODS: Expression of EMAP-2 in the placenta was investigated using reverse transcriptase-polymerase chain reaction to detect mRNA, and immunohistochemistry with an EMAP-2-specific polyclonal antiserum was carried out to detect protein. Levels of circulating EMAP-2 protein were measured in the blood of nonpregnant, normal pregnant, and preeclamptic individuals using a specific enzyme-linked immunoabsorbent assay and the molecular forms present assessed by Western blotting. RESULTS: EMAP-2 is transcribed and translated by the placenta troughout pregnancy and in preeclampsia and can be detected in the plasma of nonpregnant and pregnant individuals. Levels of circulating EMAP-2 antigen are raised in pregnancy compared with nonpregnant controls; however, levels in patients with preeclampsia are identical to those in normal pregnant individuals. CONCLUSION: While circulating levels of EMAP-2 are increased in pregnancy, there is no evidence that EMAP-2 is involved directly in the pathogenesis of preeclampsia. PMID- 10376271 TI - Effect of cocaine on intracellular calcium regulation in myometrium from pregnant women. AB - OBJECTIVE: To evaluate the effect of cocaine on intracellular free calcium ([Ca2+]i) regulation in human myometrial cells by determining the sources of Ca2+ it might mobilize, as well as assess the role cocaine might play in the catecholamine's effect on the cell's [Ca2+]i. METHODS: Primary culture of myometrial cells from pregnant women was used as an experimental model. [Ca2+]i relative changes in response to cocaine and norepinephrine were measured with fura-2 fluorometry and analyzed by means of one-way analysis of variance. RESULTS: Cocaine alone (10(-8) to 10(-3) mol/L) increased [Ca2+]i by up to 43 +/- 18% over basal level in a dose-dependent manner. Norepinephrine also elevated [Ca2+]i in a concentration-dependent manner (202 +/- 24% over basal level at 10( 4) mol/L). The norepinephrine-evoked increase was inhibited in Ca(2+)-free media by 48%, whereas the cocaine response was not affected. The Ca(2+)-channel antagonist nifedipine caused decrease in the [Ca2+]i response to 10(-5) mol/L of norepinephrine by 84%, whereas the [Ca2+]i rise to 10(-5) mol/L cocaine was not significantly changed. Inhibitor of the sarcoplasmic reticulum Ca2+ pump, thapsigargin, completely blocked cocaine-evoked increases in [Ca2+]i, whereas norepinephrine responses were greatly reduced. At the same time, cocaine (10(-8) to 10(-3) mol/L) did not potentiate norepinephrine-evoked Ca2+]i increases in the cells. CONCLUSION: These results indicate that cocaine increases [Ca2+]i in pregnant human myometrial cells, primarily by stimulating release of Ca2+ from intracellular stores rather than by direct stimulation of Ca2+ influx. PMID- 10376272 TI - Peritoneal fluid cytokine and eicosanoid levels and their relation to the incidence of peritoneal adhesion. AB - OBJECTIVE: To determine the peritoneal fluid content of several cytokines and eicosanoids with inflammatory, anti-inflammatory, anti-fibrotic, and fibrotic activities, and to assess the relationship of these levels with the incidence of peritoneal adhesions. METHODS: Peritoneal fluids were collected from 30 subjects with adhesions (n = 22) or with normal pelvic anatomy (n = 8), and the level of interleukin-1 beta (IL-1 beta), tumor necrosis factor alpha (TNF-alpha), interleukin-10 (IL-10), interferon gamma (IFN-gamma), transforming growth factor beta-1 (TGF-beta 1), granulocyte macrophage-colony stimulating factor (GM-CSF), prostaglandin E2 (PGE2), leukotriene B4 (LTB4), and thromboxane B2 (TXB2) were determined by enzyme-linked immunosorbent assay (ELISA) and radioreceptor assay. RESULTS: The peritoneal fluid content of these factors varied considerably, with low levels of IL-1 beta, TNF-alpha, IL-10, IFN-gamma, and GM-CSF. Only IFN-gamma levels were significantly lower in subjects with adhesions compared with the normal group (P < .05). The levels of total (latent + active) and active TGF-beta 1 were higher than those of other cytokines assayed and were significantly higher in subjects with adhesions compared with the normal group (P < .05). The peritoneal fluid content of PGE2, TXB2, and LTB4 was significantly higher than that of the cytokines and was higher, but not significantly so, in subjects with adhesions compared with normal subjects (P = .06). CONCLUSION: Although the effect of length of time since the adhesions were formed is not known, the results indicate that peritoneal fluid content of these cytokines and eicosanoids, with the exception of IFN-gamma and TGF-beta 1, does not correlate with the presence of peritoneal adhesions. PMID- 10376273 TI - Collagen phagocytosis by human extravillous trophoblast: potential role in trophoblastic invasion. AB - OBJECTIVES: To study collagen phagocytosis by human extravillous trophoblast. METHODS: First-trimester extravillous trophoblastic cell lines and primary trophoblast cell preparations were cultured in vitro with collagen-coated fluorescent latex beads and fluorescent-labeled collagen. Confocal microscopy was used to demonstrate internalization of collagen and beads. The effect of cytochalasin B, temperature, metabolic inhibitors, and cytokines was studied by culturing trophoblast cells with tritiated collagen. Acridine orange was used to stain for lysosomal compartments, and histochemical methods were used to demonstrate acid phosphatase in trophoblast cells. RESULTS: Both cell lines and primary culture cells internalize collagen and beads. Confocal microscopy unequivocally localized the phagocytosed material to the intracellular compartment. Inhibition by cytochalasin B and culture at 4C of uptake of [3H] collagen suggested that the process was phagocytosis. Cytokines and growth factors did not affect phagocytosis. Lysosomal compartments and acid phosphatase appear to colocalize. CONCLUSIONS: The continuous remodeling and turnover of collagen which occur in a wide variety of tissues under both physiologic and pathologic conditions are thought to be mediated by two pathways: one external (involving release of proteolytic enzymes), and the other internal (involving phagocytosis). Similar remodeling events are likely to occur during trophoblast invasion. Although current views emphasize the importance of the extracellular pathway, we postulate, on the basis of our findings, that both pathways are used, with the internal pathway probably being dominant. We hypothesize that the proteolytic enzymes (extracellular pathway) disrupt collagen matrices, thereby facilitating phagocytosis. It is teleologically sound to conceive of a dominant intracellular pathway, as it allows for more precise control of the process of invasion and is economical, as the products of collagen degradation can be used as energy sources or building blocks. PMID- 10376274 TI - The effect of diet on blood vitamin K status and urinary mineral excretion assessed by a food questionnaire. AB - To assess Vitamin K (VK) status by food questionnaire, healthy young males (32) and females (9) were given a food list of 50 VK rich foods (identified in the 4th edition standard tables of food composition in Japan). After checking the food names and approximate amount eaten for 7 days, early morning blood and urine samples were collected. Prothrombin and hepaplastin was tested and plasma protein induced by VK absence factor II (PIVKA-II), osteocalcin, and calcium, phosphorous and magnesium in plasma and urine were determined. Participants who have a habit of eating natto, a traditional Japanese food very rich in Vitamin K, VK were excluded, and lowest and highest VK consumers were compared (males; lowest 7 vs. highest 7, females; 3 vs. 3). Plasma PIVKA-II levels, and urinary calcium and magnesium excretion of the lowest was significantly higher, but urinary phosphorus was lower, than that of the highest in females. In the natto eaters, daily mean VK intakes and hepaplastin test results of natto eaters were significantly higher, but urinary calcium excretions were lower than that of non natto eaters in males. These results suggest that Daily VK intake estimated from a questionnaire, is well related to real VK status, and also that natto is a good dietary source of vitamin K. PMID- 10376275 TI - Infant feeding practices in a deprived environment: a concern for early introduction of water and glucose D water to neonates. AB - The main objective of this study was to inquire from lactating mothers whether they were fully or partially practising exclusive breastfeeding in the first six months postnatally. Time of initiation of breast and complementary feedings, types of feeds and reasons for giving other feeds to infants apart from breast milk were also examined. The data were collected by structured pretested questionnaire. Of the 200 nursing mothers interviewed, 103 (51.5%) and 77 (38.5%) reported to have given water and glucose D water to neonates respectively within the first week of life. Sieved cornpap was the popular weaning diet. Time of introducing complementary feeding to infants, and nursing mothers' educational levels, were highly significantly related (P = 0.005). Surprisingly, none of the nursing mothers listed infant formula as one of the complementary feeds. It is concluded that there is a strong need to correct this unnecessary practice of giving water and glucose D water to neonates to prevent thirst and Jaundice respectively. The correction should commence with health workers and then the nursing mothers. PMID- 10376276 TI - McCarrison, farmyard manure and the future. PMID- 10376277 TI - The significance of dietary boron, with particular reference to athletes. AB - Ergogenic substances and synthetic steroids have a wide spread use, particularly among non-professional athletes. To avoid the side-effect of drug abuse, it is suggested that the key to success is a proper athletic nutrition. It is a balanced intake of nutritional wholesome foods that contain a proper blend of essential nutrients. Knowledge of human physiology and nutrition has increased greatly, and so has application of dietary alterations and supplementation with specific nutrients. Modulation of dietary composition and/or supplementation with specific nutrients with the intent of improving human physical performance is a working definition of nutritional ergogenic aids. Boron is a trace element nutrient, and recently its supplements have been shown to increase the concentration of plasma steroid hormones. In a single blind cross-over trial, it resulted in a significant increase in plasma 17-B estradiol (E2) concentration (P < 0.004) and there was a trend for plasma testosterone (T) levels to be increased. The ratio of E2/T increased significantly. However, there was no perturbation in plasma lipids. Furthermore, the effect of boron on steroidogenesis and its mechanism was also investigated in two more studies conducted on adult male rats. The elevation of endogenous steroid hormones as a result of boron supplementation suggest that boron may be used as an ergogenic safe substance for athletes which should be further investigated. PMID- 10376278 TI - Preliminary results on the effect of tetracycline on the embryogenesis and symbiotic bacteria (Wolbachia) of Dirofilaria immitis. An update and discussion. AB - The distribution and phylogeny of Wolbachia in filarial species suggests that these endosymbiotic bacteria may be important in the biology of their filarial hosts. An experiment to falsify this hypothesis would be to treat filarial worms with antibiotics which are active against intracellular bacteria. Indeed, it has already been shown that tetracycline treatment inhibits development in a model filarial species (Brugia pahangi) at different stages of the life cycle, in both mosquito and mammalian hosts. Here we discuss these previous data and present new results on the effect of tetracycline on the embryogenesis of the canine filaria Dirofilaria immitis. PMID- 10376279 TI - Different patterns of intestinal helminth infection among young workers in urban and rural areas of Alexandria Governorate, Egypt. AB - The process of urbanisation taking place in most developing countries is creating favourable conditions for an increase in prevalence of infections, especially with intestinal parasites, in the marginal areas of the towns. The present study was implemented in 1996 to assess the varying prevalence and intensity of infection among young workers in urban and rural areas of the same Governorate (Alexandria, Egypt). The sample comprised 408 male subjects, 8 to 19 years of age, in various occupations: 308 from urban areas, 67 from an industrialised village close to the desert, and 33 from a rural village. A quantitative diagnosis of intestinal helminth infections was made using the Kato-Katz technique, with a double reading of each slide. The results showed a higher prevalence (> 50%) and intensity of infection (indirectly measured as number of eggs per gram of faeces) than in previous studies. Furthermore, a higher prevalence and intensity of infection with Ascaris lumbricoides and Trichuris trichiura was detected in urban districts, as compared to rural areas. This difference was statistically significant. High crowding index, latrine shared with other families and no piped water inside the household, were more common in urban areas as compared with rural settlements and also associated with a higher intensity of infection by soil-transmitted helminths. The trend toward urbanisation seems to have caused deterioration of living conditions and sanitation standards in some areas of Alexandria city, with the most vulnerable people experiencing an increase in intestinal parasitic infections. PMID- 10376280 TI - Chronosexuality of Plasmodium species of Central African Muridae. AB - A host harbouring many parasite species of the same genus is a phenomenon frequently observed in numerous parasitic infections. This is the case for the Plasmodium parasites of Muridae in Central Africa, where three different parasite species are found in the same rodent host species. It is highly likely that these three Plasmodium species are transmitted simultaneously by the same vector. We and others have shown that the maturation periods of the various asexual and sexual stages in the rodent, differ amongst the three parasites. In this article we propose that these differences are the product of complex adaptations which result, for all three Plasmodium species, in a maximum peak of infectivity to the insect vector occurring around 3 a.m., the period of highest activity of the nocturnal host rodent. PMID- 10376281 TI - Malaria, schistosomiasis, and intestinal helminths in relation to microdams in Tigray, northern Ethiopia. AB - A survey was undertaken in Tigray, Northern Ethiopia, to assess the prevalence of malaria, schistosomiasis, and intestinal helminths in relation to microdams. The survey took place from March to June 1995, during the dry season, at 41 microdams. At each site the village nearest the dam (within thirty minutes walk) was selected, ten households were randomly chosen, and all family members were examined for malaria and intestinal parasites. The overall study sample was 2271 people, of all age groups. Plasmodium falciparum infection was documented in four villages (at 10% of microdams); prevalence was 1.2% (range 0-20% by village). Larvae of Anopheles gambiae s.l. were found at one microdam. Infection with intestinal schistosomiasis was documented in 20 villages (at 49% of microdams), and one third of those infected had moderate to heavy infections. Biomphalaria species, the intermediate host snails of Schistosoma mansoni, were found at 16 microdams (39%), and snails infected by mammalian cercariae were found in one locality. Infections with soil-transmitted nematodes were prevalent: hookworm was detected in more than two thirds of the villages, and Ascaris lumbricoides and Trichuris trichiura were present in almost half of the villages. Out of 2078 stool examinations, the prevalence of S. mansoni infection was 7.2% (range 0-48% by village), of A. lumbricoides 2.3% (range 0-31%), of T. trichiura 2.4% (range 0 21%), and of hookworm 8.9% (range 0-78%). The prevalence of malaria, S. mansoni and hookworm was higher at altitudes below 2000 metres above sea level. S. mansoni was more prevalent in microdams built more than 5 years before the survey, while T. trichiura was more prevalent at recently constructed microdams. The widespread distribution of schistosomiasis and intestinal helminths, and the presence of malaria infection during the dry season confirm that microdams create favourable conditions for the transmission of these parasitic diseases. Health safeguards must be incorporated into the planning, construction, and operation of microdams and irrigation systems in order to prevent or reduce these diseases. In areas with high prevalence, control activities should be intensified. PMID- 10376282 TI - Breast infection due to Dirofilaria repens: report of two new Italian cases and revision of the literature. AB - Two new cases of human dirofilariasis, occurred in women aged 46 and 52 years, respectively, both living in Lombardy (Northern Italy) are reported. Dirofilaria repens nematodes were localised in the breast in both cases. In one of them the parasite was accidentally extracted while the patient underwent a fine needle aspirate. The international literature records 30 cases of breast Dirofilariasis. Except in rare cases, the parasite was located subcutaneously in the breast, and nested in a nodule. The clinical diagnoses were consistently wrong, the nodule being diagnosed as a suspected tumor of the breast. PMID- 10376283 TI - Acanthamoeba adherence to contact lenses, removal by rinsing procedures, and survival to some ophthalmic products. AB - Unworn soft and rigid gas-permeable contact lenses were inoculated with an Acanthamoeba keratitis strain to study the protozoon's ability to adhere. Furthermore, the efficacy of the rinsing in saline on acanthamoeba removal was evaluated, as well as the amebicidal activity of five commercial cleaning/disinfecting products: hydrogen peroxide, chlorhexidine, polyaminopropyl biguanide-poloxamine, thimerosal-polyquaternium, and thimerosal-chlorhexidine. Microscopic count of cells showed that Acanthamoeba trophozoites and cysts adhered to all types of contact lenses. A significantly greater adherence of trophozoites than cysts was recorded. The rinsing in saline using a flow-method was significantly more effective than the immersion-method, particularly in removing trophozoites from rigid gas-permeable lenses. The cleaning/disinfecting solutions tested were ineffective in removing or in affecting the viability of all Acanthamoeba trophozoites or cysts in the 17 hours allotted for the experiment. The need for a better care in mechanical and physical hygiene procedures is stressed. PMID- 10376284 TI - Sense organs in post-embryonic stages of Hyalomma marginatum marginatum Koch, 1844 (Acari: Ixodida: Ixodidae). I. Tarsal sensory system. AB - Light and scanning electron microscopic studies showed the differences in morphology and in size of Haller's organ in larvae, nymphs and adults (females and males) of Hyalomma marginatum marginatum Koch, 1844. The length of the anterior pit setae increases during post-embryonic development. The localization of these setae is the same in all stages. Six setae (one porose seta, two grooved setae, two fine setae, one conical seta) contain anterior pit of various developmental stages. In nymphs and adults more numerous pores appear on the wall surface of porose seta than in the larval stage. The structure of the capsule roof also differs in various developmental stages. Haller's organ of Hyalomma m. marginatum shows great degree of morphological development which is connected with the complicated life cycle and feeding behaviour of this tick species. PMID- 10376285 TI - The fine structure of reptilian Isospora species with intranuclear endogenous development. AB - The fine structure of the endogenous stages of five species of Isospora of lacertile hosts is reviewed and compared. All species were intranuclear and were the following: I. deserti, from Agama pallida from Israel; I. cannoni, from Diporiphora australis from Queensland, Australia; and new, yet undescribed species from Hemidactylus turcicus from Israel. Heteronota binoei from Queensland, and Carlia rhomboidalis from Queensland. In young infections the intranuclear parasitophorous vacuole (PV) maintained at one point a junction zone with the nucleolemma. Finding couples of trophozoites sharing the same PV suggests a binary division (or endodyogeny?) prior to merogony. There was some overall conformity in structure among the respective stages of the different species, particularly of the gamonts. Some interspecific differences were, however, evident in the texture of the wall-forming bodies. The wall formation followed the scheme described for avian and mammalian eimeriid coccidia. The smaller size Isospora from C. rhomboidalis exhibited fine structural peculiarities and affected its host nucleus differently. More conspicuous peculiarities were evident in the differentiation process of I. cannoni meronts, involving a formation of centrally positioned large inclusions, a mitochondrial plaque-like organelle and large uniquely structured vesicular mitochondria. PMID- 10376286 TI - [Observations on subgenus Aedes (genus Aedes, Diptera, Culicidae) in northeast Italy and first italian report of Aedes geminus Peus]. AB - The paper reports some observations on the subgenus Aedes (genus Aedes, Diptera, Culicidae) in northeast Italy. Two species were collected: Ae. cinereus and Ae. geminus, the latter recorded for the first time in Italy. Morphological, ecological and biological data of the two species are presented. The identification is possible only on the male hypopigium; larvae, pupae and adult females show no differential characters. For both species, the larval breeding sites were fresh water marshes mainly within woods; preimaginal development took place twice a year, in Spring and Autumn. The females were strongly anthropophilic. No biological differences between the two species were noticed, but more data are needed to ascertain their relationships and the presence of subtle biological divergences. PMID- 10376287 TI - Parasites of Italian sea turtles. II. Loggerhead turtles (Caretta caretta [Linnaeus, 1758]). AB - Eleven out of fourteen sea turtles (Caretta caretta) stranded along the coast of the Adriatic sea (Mediterranean sea) were found infected with helminths. Seven trematode species (Rhytidodes gelatinosus, Orchidasma amphiorchis, Enodiotrema megachondrus. Pachypsolus irroratus, Pleurogonius trigonocephalus, Calicodes anthos, Plesiochorus cymbiformis) and one nematode (Sulcascaris sulcata) were found. S. sulcata was the most frequent species in the stomach and R. gelatinosus in the intestine. These species were also the most abundant. The redescriptions of C. anthos (Braun, 1899) and P. trigonocephalus (Rudolphi, 1809) Looss, 1901 are included. The accuracy of Braun's original description is questioned because it refers to specimens studied in dorsal view. PMID- 10376288 TI - Some aspects of intracellular symbiosis during embryo development of Mastotermes darwiniensis (Isoptera: Mastotermitidae). AB - All examined species of cockroaches have been shown to harbour intracellular bacteria in specialized cells (bacteriocytes) of the fat body. In termites, bacteria in specialized cells have been observed only in Mastotermes darwiniensis (Isoptera: Mastotermitidae). All of these bacteria have been assigned to the same eubacterial lineage, with the bacteria of M. darwiniensis as the sister group to the cockroach bacteria. While the main steps of the life cycle of cockroach bacteria have been described, little is known about the bacteria of M. darwiniensis. More specifically, no data are available on their behaviour during the development of this termite. Using both optical and electron microscopy methods, we examined embryos of M. darwiniensis at different developmental stages. Our results show that the integration of bacteria during the development of M. darwiniensis is implemented in the same way as in cockroaches. In particular, we observed the aggregation of a large amount of bacteria in a single mass in the yolk sac, with vitellophage-associated bacterial lysis. In cockroaches, a similar process has been described in detail for Periplaneta americana (Blattaria: Blattidae), where the bacterial mass is referred to as the transitory mycetome. The formation of a transitory mycetome could thus be regarded as an ancestral condition for cockroaches and termites. PMID- 10376289 TI - [Myasis due to Cordylobia anthrophaga (Diptera: Calliphoirdae): description of a clinical case and review of the literature]. AB - A case of myiasis due to Cordylobia anthropophaga is reported. The patient, an Italian man 44 years old, had come back from Nigeria and Ghana. He presented two nodular erythematous lesions of about 1 cm in diameter, on his back and right arm. From the lesions two third larval stage of C. anthropophaga were extracted. The authors report a short review about myiasis cases described in the scientific literature in Italy. PMID- 10376290 TI - Results of a survey to detect the mosquito Aedes albopictus in the French Riviera. AB - A programme of surveillance was initiated in 1992, in the French Riviera, to detect a possible introduction of Aedes albopictus from Italy and to prevent nuisances caused by mosquitoes in the touristic localities of the Cote d'Azur. In five years, numerous mosquito breeding places have been located. Nine species have been collected: Anopheles claviger, An. plumbeus, Aedes geniculatus, Ae. vittatus, Culex hortensis, Cx. impudicus, Cx. pipiens, Culiseta fumipennis, Cs. longiareolata but no Ae. albopictus was found. Nuisances were mainly due to hypogean populations of Cx. pipiens. Breeding places in urban sites have been controlled or suppressed. The discovery of an important larval population of An. plumbeus in an urban area might further stress the importance of this species already suspected to transmit indigenous malaria in cities. PMID- 10376291 TI - Hypoderma lineatum antigen and anti-Przhevalskiana silenus antibodies: cross reactivity and antibody kinetics in naturally infested goats. AB - To evaluate the cross-reactivity between Hypoderma lineatum antigen and anti Przhevalskiana silenus antibodies six protocols with different concentrations of antigen and different dilutions of sera and conjugate were applied. The highest cross-reaction between the H. lineatum antigen and the anti-P. silenus antibodies is given by 2 micrograms/ml of antigen concentration, 1:400 of serum and 1:10,000 conjugate dilution. The study on the kinetic development of antibodies in goats naturally infested by P. silenus and the natural course of infestation pointed out the existence of a good correlation between individual antibody kinetics and the natural evolution of the cycle of infestation. The highest antibody concentration may be registered from October through November, coinciding with the end of the migration fo the larvae inside the animal's body. In our condition, this period can be considered as a favorable sampling period for immunodiagnosis and immunoepidemiological studies of goat warble fly infestation. PMID- 10376292 TI - Both human subcutaneous dirofilariasis cases previously diagnosed in Amiens (France) are probably not autochthonous. PMID- 10376293 TI - [A case of cutaneous myiasis caused by Dermatobia hominis]. AB - Due to increasing international travelling of European population, tropical parasitic diseases may be imported more frequently in temperate countries. We describe a subject who travelled in Brasil and returned to Italy with a subcutaneous nodule containing a phase-II Dermatobia hominis larva; such larva is reported with photographic documentation. PMID- 10376294 TI - Trichomonosis of the oral cavity complicated by mycosis. AB - Protozoa and fungi of the oral cavity, although frequently occurring and connected to considerable clinical adverse effects, are as yet insufficiently known. The aims of the study were (i) to estimate the prevalence of common invasions of Trichomonas tenax and fungi, (ii) to associate the symptoms with the diagnosis of trichomonosis complicated by mycosis, and (iii) to determine trichomonacidal properties of ornidazole, tinidazole and metronidazole. A sample of 936 dentist patients with different diagnoses were included in the study. The collected material consisted of rinsings, with simultaneous application of selective media, different for protozoa or fungi cultures. Using ornidazole, tinidazole and metronidazole, we examined in vitro their influence on 30 strains of T. tenax. Among the examined patients T. tenax was found in 90 cases including 85 cases where it occurred together with fungi, on the basis of which diagnosis of trichomonosis complicated by mycosis of the oral cavity was established. It was the most frequent in patients with leukoplakia and lichen Wilsoni. We recorded a statistically significant association for T. tenax with fungi and xerostomia, burning sensation, periodontal pockets and denuded teeth. T. tenax was never found in patients with caries and with aphtha recidivans. The curves of ornidazole activity were obtained within the solution range of 130-4350 micrograms/ml; the curves of tinidazole activity within the concentration range of 500-16870 micrograms/ml; metronidazole in the highest concentration killed from 5 to 100% of the population of all strains. PMID- 10376295 TI - Presence of Dirofilaria repens and an insect immunocyte (plasmatocyte) in a human subcutaneous nodule, induced by a mosquito bite. AB - It is well known that the nematode Dirofilaria repens is transmitted to humans by vector mosquito bite. Examination of a fine needle aspiration biopsy drawn from a month-old nodule on the chest of a woman, residing in Garlasco, province of Pavia, Northern Italy, revealed the presence of not only one immature female of D. repens, but also some scattered cells that we believe to be mosquito's blood cells, plasmatocytes (immunocytes). We presume that plasmatocytes were carried into the bite wound with the mosquito's hemolymph that had oozed from a rupture in its mouthparts during feeding. Because Aedes albopictus recently colonized certain areas in the above region, we suspect that the nodule resulted from the bite of this mosquito. PMID- 10376296 TI - [Classification of Anopheles claviger (Diptera, Culicidae) in north-eastern Italy]. AB - The paper reports the results of a taxonomical study and some reservations on the ecology and bionomics of the Anopheles claviger complex of north-eastern Italy. In 1996 and 1997, 18 samples of larvae and pupae were collected in 15 localities of Friuli-Venezia Giulia and eastern Veneto regions, from sea level up to 200 m a.s.l.; 870 larvae and 114 pupae were examined and identified by morphological criteria. All samples and all individuals were assigned to the species An. claviger s. str. Abnormal setae 1-II were detected in some larvae; significant statistical differences were recorded among some samples for the larval characters setae 1-II, 2-IV and 2-V. The breeding sites were mainly originated by springs, which are abundant in the region. The species showed two generations per year, with overwintering as a larva. PMID- 10376297 TI - [Description of Aedes (Ochlerotatus) coluzzii n. sp. (Diptera, Culicidae), twin A species of the detritus complex]. AB - Reports of a wide variation in space and time of the frequency of autogeny in Aedes (Ochlerotatus) detritus (Haliday, 1833) of the Camargue (Delta of the Rhone, Bouches-du-Rhone, Gard, France) led us to make an enzymatic analysis of male and female adults from different larval biotopes. The study showed the existence of two genetically distinct, sympatric populations which are morphologically indistinguishable. By diagnostic enzymes monomorph Got-2, Gpd, the grouping of the individual into two subgroups satisfies a Hardy-Weinberg equilibrium for the polymorphic enzymes. It is concluded that Ae. detritus is composed of a complex of two sibling species provisionally designated by the letters A (Got-2RR, GpdCC) and B (Got-2LL, GpdBB). In the present article, we retrace the history of the binomen Ae. detritus since the original description (sub nom. Culex detritus) to the split into the detritus complex. Certain ecophysiological (steno-eurygamy) and chorological (bioclimatic gradients N-S) criteria show that the sibling species B should be assigned to the taxon described in UK (Holywood, County Down, Ireland) by A.H. Haliday. Species A is here named Aedes (Ochlerotatus) Coluzzii n. sp. PMID- 10376298 TI - Comparison of milk protein allele frequencies in Nordic cattle breeds. AB - Allele frequencies at four milk protein loci were studied in five modern and 17 old Nordic cattle breeds in order to reveal variants that are characteristic for these populations. The B allele of CSN3, which has been associated with improved manufacturing properties of milk, showed significantly lower frequencies in modern production breeds than in old breeds of interest for conservation purposes. Characteristic frequencies of CSN1S1 (C), CSN2 (A2) and CSN3 (B) were found in Icelandic cattle, Swedish Mountain cattle, Northern Finncattle and Western Fjord cattle, which indicate a common origin of these populations. Further comparisons of allele frequencies in old Nordic breeds suggest sorting of these breeds into two groups with a northern and southern geographic location. PMID- 10376299 TI - Major histocompatibility complex (MHC) related cDNA probes associated with antibody response in meat-type chickens. AB - The major histocompatibility complex (MHC) region was examined as a set of candidate genes for association between DNA markers and antibody response. Intercross F2 families of chickens were generated from a cross between high (HC) and low (LC) Escherichia coli(i) antibody lines. Restriction fragment length polymorphism (RFLP) analysis was conducted by using three MHC-related cDNA probes: chicken MHC class IV (B-G), chicken MHC class I (B-F), and human MHC linked Tap2. Association between RFLP bands and three antibody response traits (E. coli, sheep red blood cells and Newcastle disease virus) were determined by two methods: by statistically analyzing each band separately and also by analyzing all bands obtained from the three probes by using multiple regression analysis to account for the multiple comparisons. The MHC class IV probe was the highest in polymorphisms but had the lowest number of bands associated with antibody response. The MHC class I probe yielded 15 polymorphic bands of which four exhibited association with antibody response traits. The Tap2 probe yielded 20 different RFLP bands of which five were associated with antibody production. Some Tap2 bands were associated with multiple antibody response traits. The multiband analysis of the three probes' bands revealed more significant effects than the analysis of each band separately. This study illustrates the efficacy of using multiple MHC region probes as candidate markers for quantitative trait loci (QTLs) controlling antibody response in chickens. PMID- 10376300 TI - Phylogenetic relationships of Cheju horses to other horse breeds as determined by mtDNA D-loop sequence polymorphism. AB - Historical records suggest that horses inhabiting the island of Cheju in Korea are descendants of Mongolian horses introduced in 1276. Other studies, however, suggest that horses may have been present on the island prior to the Mongolian introduction. To determine the origin of the Cheju horses we used a phylogenetic analysis of sequences of the mitochondrial DNA (mtDNA) D-loop region, including tRNA Pro and parts of tRNA thr and tRNA Phe sequences (1102-bp excluding the tandem repeat region). Maximum parsimony and neighbor-joining trees were constructed using sequences determined for seven Cheju, four Mongolian, one Przewalskii and two Chinese Yunnan horses, and published sequences for one Swedish and three Thoroughbred horses. Donkey mtDNA was used as an outgroup. We found that the mtDNA D-loop sequence varies considerably within Mongolian, Cheju and Thoroughbred horse breeds, and that Cheju horses clustered with Mongolian horses as well as with horses from other distantly related breeds. On the basis of these findings we propose that horses on Cheju Island are of mixed origin in their maternal lineage, and that horses may have existed and been traded on the island before the Mongolian introduction. PMID- 10376301 TI - A PCR method for typing B-L beta II family (class II MHC) alleles in broiler chickens. AB - Certain haplotypes of the major histocompatibility (B) complex are strongly associated with resistance or susceptibility to several infectious diseases in Leghorn chickens. Identification of chicken haplotypes based on the nucleotide sequence of B complex loci could provide more precise identification of haplotypes than traditional serological methods. We report the development and application of polymerase chain reaction with sequence specific primers (PCR-SSP) to type broiler chicken B haplotypes based on the DNA sequence of B-L beta II family genes. Five well-defined standard B haplotypes from White Leghorns and 12 recently characterized B haplotypes from a broiler breeder line were used to develop the test system. The B-L beta II family loci were amplified from genomic DNA by B-L beta II family specific primers and then characterized by PCR-SSP. In total, ten pairs of primers, derived from the sequences of expressed B-L beta II family alleles, were used in the PCR typing test to discriminate the chicken B haplotypes identified previously by serological means. The PCR-SSP showed that each haplotype had a different amplification pattern, except those haplotypes known or suspected to have the same B-L beta alleles. Cloning and sequencing of the family specific PCR products indicated that two loci in the B-L beta II family, presumably B-L beta I and B-L beta II, were amplified. Finally, B-L beta PCR-SSP typing was used in combination with B-G RFLP analyses to characterize unusual (variant) B serotypes; the results indicate that some of these are natural recombinants within the B complex. PMID- 10376302 TI - Conserved nucleotide differences and subfamily structure of porcine short interspersed elements. AB - Interspersed elements are ubiquitous in the genomes of higher eukaryotes and account for over a third of the genomic DNA (Smit 1996). In swine the short interspersed elements, SINEs or PREs (porcine repetitive elements), have been found in a number of introns and 3' untranslated regions of different genes. However, compared to human Alu repeats the number of available PRE DNA sequences is still limited. In this study we have compared 85 PREs selected from DNA sequence database entries. The PREs were aligned and for each nucleotide position the relative frequencies of the four bases were calculated. A consensus sequence was derived from the first base usage. Similar to studies of SINEs in other species, the analysis showed that most mutations in PREs occur at CpG dinucleotide hot spots. The position variability for the two most frequent bases shows a bimodal distribution. The analysis suggests that the porcine SINEs can be divided into three major subfamilies sharing conserved nucleotide similarities. PMID- 10376303 TI - High resolution mapping and identification of new quantitative trait loci (QTL) affecting susceptibility to Marek's disease. AB - Marek's disease (MD) is a lymphoproliferative disease of chickens that costs the poultry industry approximately $1 billion annually. Genetic resistance to MD is gaining increased attention to augment vaccinal control as disease outbreaks occur more frequently. Previously, analysis of a 272 F2 White Leghorn resource population measured for many MD traits and genotyped for 78 microsatellite markers revealed two and four quantitative trait loci (QTL) with significant and suggestive association, respectively, to one or more MD associated traits. Additional genetic markers have since been scored on the MD resource population to increase QTL resolution and genome coverage. Saturation of four of the QTL regions with 17 markers revealed five new QTL while 32 markers extended the genome coverage by 400 + CM and uncovered three more QTL. QTL analysis by single point and interval mapping algorithms agreed well when marker saturation was approximately 20 CM or less. Currently 127 genetic markers cover approximately 68% of the genome that contain up to 14 MD QTL associated to one or more MD trait; seven at the significant level and seven at the suggestive level. Individually each QTL accounts for 2-10% of the variation and, in general, resistance was dominant although the resistant allele may come from either parental line. This study suggests that a limited number of genomic regions play a major role in the genetic control of MD resistance. Markers linked to these loci may be useful for selection of MD resistant stock by the poultry industry following verification of the association within their breeding populations. PMID- 10376304 TI - A search for quantitative trait loci for milk production traits on chromosome 6 in Finnish Ayrshire cattle. AB - Cattle chromosome 6 was scanned with 11 markers, ten microsatellites and the casein haplotype, to identify quantitative trait loci (QTLs) affecting the following milk production traits: milk yield, fat percentage, fat yield, protein percentage and protein yield. Twelve Finnish Ayrshire half-sib families with a total of 480 sons were genotyped and used in a grand-daughter design. Interval mapping was performed with a multiple-marker regression approach with a one-QTL and a two-QTL model, and the significance threshold values were determined empirically using a permutation test. Across-family analysis with the one-QTL model revealed an effect on protein percentage (P < 0.05) and on milk yield (P < 0.05). The analysis with the two-QTL model identified significant effects (P < 0.05) on protein percentage, milk yield, and fat yield. Comparing these two cases, the results suggest the existence of two QTLs on chromosome 6 with an effect on milk production traits. One of the QTLs was located around the casein genes. As the other QTL was similar in location and effect to a QTL found previously in Holstein-Friesians, an identity-by-descent approach could be applied to fine map this region. PMID- 10376305 TI - The AB blood group system in wild felids. AB - The blood type of 131 non-domesticated felids belonging to 26 felid species was surveyed in this study. Based upon a tube hemagglutination assay established for domestic cats, 80% of felids had type-A, 18% type-B, and 2% type-AB blood. Felids in the Puma group and African and Asian golden cats had blood type B, whereas all other species were found to have blood type A. Two cheetahs and one bobcat had type-AB blood. Red cell glycolipids analysed by high performance thin layer chromatography revealed a similar ganglioside pattern in wild cats as reported in domestic cats. Independent of the AB blood group system, incompatible blood crossmatch reactions were detected between different felid groups. In conclusion, wild felids display the same AB-erythrocyte antigens as domestic cats, and the same blood typing procedures can be applied for wild and domestic felids. PMID- 10376306 TI - A convenient and efficient microsatellite-based assay for resolving parentages in dogs. AB - We present a practical and efficient parentage analysis test for dogs. The method makes use of ten polymorphic microsatellite markers combined in three multiplex PCR reactions. All three multiplexes can be performed under the same cycling conditions and using a multiple-dye detection system the PCR-products can be pooled for electrophoresis. Amplifications in four different thermal cyclers produced easily interpretable results throughout demonstrating the repeatability of the assay. In order to evaluate the method's efficiency a total of 100 dogs from four different breeds were typed. Assuming one known parent, exclusion probabilities ranging from 99.34% to 99.93% were attained. PMID- 10376307 TI - Identification of abundant and informative microsatellites from shrimp (Penaeus monodon) genome. AB - Microsatellites were isolated from P. monodon genomic libraries by direct sequencing of recombinant clones without probe screening. Forty-nine out of 83 clones sequenced contained 99 microsatellite arrays of three or more repeats. When five or more and ten or more repeats were considered, 28 and 14 microsatellites were detected, respectively. The 99 microsatellites were classified as perfect (75%), imperfect (6%), compound perfect (3%) and compound imperfect (16%). The abundance of di-, tri-, tetra- and hexanucleotide repeats were 67%, 20%, 9% and 3%, respectively. The dinucleotide repeats included 36 (CT)n, 31 (GT)n, 17(AT)n and 3 (CG)n. One octanucleotide repeat (ATTTATTC)5 was found within a large repeat sequence. Optimal annealing temperatures were determined for PCR using 11 primer sets encompassing 15 microsatellites. Ten primer sets provided successful amplifications with allele sizes generally ranging from 139 to 410 bp. All these primers amplified polymorphic loci with PIC values ranging from 0.63 to 0.96. Two primer sets amplified additional bands which can easily be distinguished from the bands of the main locus. Three out of 10 P. monodon microsatellites also amplified alleles in P. vannamei. The abundance and informative nature of P. monodon microsatellites and their potential for cross-species amplification make them useful for genetic studies. PMID- 10376308 TI - Associations of the bovine major histocompatibility complex DRB3 (BoLA-DRB3) with production traits in Canadian dairy cattle. AB - Associations of two alleles of the bovine major histocompatibility complex DRB3 gene (BoLA-DRB3) with lowered somatic cell score (SCS) and occurrence of disease (BoLA-DRB3.2* 16 and *23, respectively) have previously been documented. The objective of this study was to evaluate potential relationships between BoLA-DRB3 alleles with production traits, namely 305-day milk, milk fat and milk protein yield, in a population of Canadian dairy cattle (Holstein, n = 835 and Jersey, n = 66) over the course of two lactations. No significant associations were detected between BoLA alleles and production traits in Jerseys. In Holsteins, alleles *16 and *23 also did not show associations with production traits but allele *8 was significantly associated with increased 305-day milk, fat and protein yields in the previous lactation (the lactation prior to immunization with a gram negative core antigen vaccine), and with increased protein production in the subsequent (with reference to the time of immunization) lactation. Allele *22 was associated with decreased milk and protein yield in both previous and subsequent lactations. Therefore, it can be concluded that increasing or decreasing the frequency of BoLA alleles *16 and *23 to reduce SCS or increase resistance to mastitis in this population would not have adverse effects on production in this population, and that certain BoLA alleles (*8 and *22) are associated with altered production traits in Canadian Holsteins. PMID- 10376309 TI - PL44, a cosmid containing microsatellite INRA242 (DXS39), maps to bovine chromosome Xq25. PMID- 10376310 TI - Clarification of the order of acrosin and aconitase 2 genes on the physical and linkage maps of porcine chromosome 5. PMID- 10376311 TI - Two polymorphic dinucleotide repeats in rainbow trout (Oncorhynchus mykiss). PMID- 10376312 TI - Linkage mapping of genes encoding bone morphogenetic proteins 1, 4 and 5 in sheep. PMID- 10376313 TI - A new CSN3 allele in Bos indicus cattle is characterised by MspI PCR-RFLP. PMID- 10376314 TI - MX1 maps to cattle chromosome 1. PMID- 10376316 TI - Eight microsatellite markers for South American camelids. PMID- 10376317 TI - Polymorphism in exon 10 of the bovine GHR gene detected by PCR-DGGE. PMID- 10376315 TI - The porcine poly(rC)-binding protein 2 (PCBP2) gene maps to chromosome 5. PMID- 10376318 TI - An AluI PCR-RFLP in the bovine survival motor neuron gene (SMN). PMID- 10376319 TI - 99mTechnetium labelled Escherichia coli. AB - Samples of a culture of unlabeled Escherichia coli were incubated with different concentrations of stannous chloride for various time periods. 99mTc (26.0 MBq) was added to each preparation and the results showed a labelling yield of 98% for E. coli. Since the bacterial viability of 99mTc-E. coli and E. coli did not show any statistical differences, these results demonstrate that labelling of E. coli with 99mTc does not modify the bacterial viability, and the radiolabelled bacteria may be a good model to study bacterial translocation. PMID- 10376320 TI - Preparation of [186Re]Re-DMSA and its bio-distribution studies. AB - 99mTc(V)-DMSA is widely used for imaging medullary carcinoma and hence 186/188Re(V)-DMSA is suggested as a potential agent for treating medullary carcinoma. In the present paper, we report the work carried out for the preparation of [186Re]Re(V)-DMSA and it's bio-distribution studies in Wistar rats. The complex was prepared by reducing 186Re (100 micrograms, 0.54 microM, approximately 150 MBq) in the presence of DMSA (2 mg, 11 microM) with stannous chloride (0.4 mg, 2.2 microM) in acidic medium at pH 2. The reaction was taken to completion by heating the complex in a boiling water bath for 30 min. Bio distribution studies carried out revealed that pharmacological behaviour of 186Re(V)-DMSA is similar to that of 99mTc(V)-DMSA except that the kidney uptake is marginally higher. The kidney uptake reduced significantly when the pH of the complex was adjusted to 8 prior to injection. The in vitro stability studies of this complex suggest that the product formed is stable and could be used for clinical trials. PMID- 10376321 TI - Preparation, stability studies and pharmacological behavior of [186Re]Re-HEDP. AB - 99mTc-HEDP is widely used as a bone imaging agent and its Re analog [186Re]Re HEDP is now well established as a therapeutic radiopharmaceutical for palliation of pain due to bone metastases. In the present paper, we report the work carried out for the preparation of stable 186Re-HEDP which retains RC purity up to 5 days when stored at 4 degrees C. 186Re was prepared by irradiation of natural Re metal at a flux of 3 x 10(13) neutrons/cm2/s for seven days and processed after a cooling period of four days. The specific activity of 186Re formed was approximately 35 mCi/mg. A complex with RC purity > 98% could be prepared by varying the reaction conditions. By carefully optimizing the reaction and storage conditions, a complex which was stable for over 4 days could be synthesized. Bio distribution studies carried out in rats revealed approximately 30% bone uptake of 186Re-HEDP at 3 h postinjection which remained almost constant for 48 h, at which time there was negligible activity in other organs. PMID- 10376322 TI - Natural radioactivity of Indian building materials and by-products. AB - Conventional building materials and by-products from coal power plants which are being used or have the potential for use in buildings in India were analysed for natural radioactivity due to the presence of 226Ra, 232Th and 40K using gamma-ray spectroscopy. The materials examined in this work showed the radioactivity levels below the limit estimated from radium equivalent activity the criterion formula for gamma-activity suggested for acceptable radiation doses attributable to building materials in OECD countries. PMID- 10376323 TI - Radon exhalation rate and uranium estimation in rock samples from Bihar uranium and copper mines using the SSNTD technique. AB - Widespread uranium mineralization is associated with copper, nickel and other sulphides in the Singhbhum shear zone developed at the northern margin of the Singhbhum craton in the state of Bihar of India. The south-eastern part of the shear zone between Surda-Mosabani-Badia is rich in copper mineralization while the central part between Jaduguda-Bhatin-Nimdih and Narwapahar-Garadih-Turamdih is enriched in uranium. In the present study, trace uranium concentration in geological samples from the Mosabani copper mine and the Narwapahar and Jaduguda uranium mine areas have been determined using fission track registration technique. For the measurement of the radon exhalation rate, the 'can technique' using alpha sensitive LR-115 type II plastic track detectors were used. Uranium concentrations were found to vary from 1.5 to 2097.9 ppm whereas the radon exhalation rate varied from 0.2 to 19.2 Bq m-2 h-1. The values of radon exhalation rate from crushed rock and soil samples are found to correspond with the measured values of uranium in the corresponding samples. A positive correlation has been found between radon exhalation rate and uranium concentration in the samples. The linear coefficients are found to be 0.40, 0.98 and 0.95 in the Mosabani, Narwapahar and Jaduguda mine areas respectively. High values of radon exhalation in subsurface mines like Jaduguda (depth approximately 800 m) and Mosabani (depth > 1000 m) seem to emphasize the need for adequate ventilation for the removal of radon and its progenies from the mines. PMID- 10376325 TI - Heavy metals and rare earth elements source-sink in some Egyptian cigarettes as determined by neutron activation analysis. AB - Heavy metals and rare earth elements in two types of cigarettes were studied. The contents of trace elements were determined by using delayed neutron activation analysis. In the present study 11 elements have been detected in popular and fine brand cigarettes marketed in Egypt. Evaluation of these elements with their potential hazards for smokers is briefly discussed. The material balance (source and sink) for each element was determined. Also the ratio of element recovery to the total amount was assessed. PMID- 10376324 TI - Distribution coefficients (Kd's) for use in risk assessment models of the Kara Sea. AB - As a prerequisite for most evaluations of radionuclide transport pathways in marine systems, it is necessary to obtain basic information on the sorption potential of contaminants onto particulate matter. Kd values for use in modeling radionuclide dispersion in the Kara Sea have been determined as part of several international programs addressing the problem of radioactive debris residing in Arctic Seas. Field and laboratory Kd experiments were conducted for the following radionuclides associated with nuclear waste: americium, europium, plutonium, cobalt, cesium and strontium. Emphasis has been placed on two regions in the Kara Sea: (i) the Novaya Zemlya Trough (NZT) and (ii) the mixing zones of the Ob and Yenisey Rivers (RMZ). Short-term batch Kd experiments were performed at-sea on ambient water column samples and on samples prepared both at-sea and in the laboratory by mixing filtered bottom water with small amounts of surficial bottom sediments (particle concentrations in samples = 1-30 mg/l). Within both regions, Kd values for individual radionuclides vary over two to three orders of magnitude. The relative particle affinities for radionuclides in the two regions are americium approximately equal to europium > plutonium > cobalt > cesium > strontium. The values determined in this study agree with minimum values given in the IAEA Technical Report [IAEA, 1985. Sediment Kd's and Concentration Factors for Radionuclides in the Marine Environment. Technical Report No. 247. International Atomic Energy Agency, Vienna.]. Given the importance of Kd's in assessments of critical transport pathways for radionuclide contaminants, we recommend that Kd ranges of values for specific elements rather than single mean values be incorporated into model simulations of radionuclide dispersion. PMID- 10376326 TI - Effect of Peumus boldus on the labeling of red blood cells and plasma proteins with technetium-99m. AB - Peumus boldus is used in popular medicine in Brazil. The influence of Peumus boldus on the labeling of red blood cells and plasma proteins with 99mTc was studied. Stannous chloride and 99mTc pertechnetate were incubated with blood and a tincture of Peumus boldus. Aliquots of plasma and blood cells were isolated from the mixture and treated with trichloroacetic acid (TCA). After separation, analysis of the soluble and insoluble fractions showed a rapid uptake of the radioactivity by blood cells in the presence of the drug, whereas there was a slight decrease in the amount of 99mTc radioactivity in the TCA-insoluble fraction of plasma. PMID- 10376327 TI - EPR dose reconstruction for bone-seeking 90Sr. AB - The results of the EPR dose reconstruction in calcified tissues of dog injected with 90Sr are presented. It has been established that there is no essential difference in the values of doses absorbed in tooth tissues of teeth in symmetric positions in the mouth, whereas a significant difference occurs in the values of absorbed doses in teeth in non-symmetric positions. In the case of 90Sr internal exposure the dose reconstruction in crown dentine plays an important role. It has been found that its quantity is close to the dose in diaphyseal cortical bone of the femur, dose at the endosteal bone surface and in femural fatty marrow. The fact that these values exceed doses absorbed in tooth enamel points out the predominant contribution of internal exposure. The highest absorbed doses have been observed in metaphyseal trabecular femur bones, tooth alveolar bone walls, and cortical and trabecular vertebra that can be considered as suitable candidates for biomarkers of internal 90Sr exposure for post-mortal autopsy. The satisfactory correlation has been found between the doses reconstructed in calcified dog tissues and the doses measured by EPR in alanine dosimeters fixed in (or nearby) the sites of autopsy of bones/teeth. The experiments provide support for the view that EPR retrospective dosimetry with calcified tissues for internal exposure is unique in providing useful information on the doses obtained. PMID- 10376328 TI - Production of radioactive endovascular stents by implantation of 133Xe ions. AB - A coronary stent was made radioactive by implantation of 133Xe ions for the purpose of suppressing the renarrowing of the part of blood vessel in which the stent is implanted. Electrons of relatively low energies emitted in the decay of 133Xe may give an antiproliferative effect of ionizing radiation to the intimal cells within a limited range of 1 mm. A 133Xe+ beam accelerated at 40 or 60 keV was directed to several stainless steel stents mounted on a target-holder table that could revolve and move up and down to distribute the 133Xe+ ions within a stent as well as among the stents. The radioactive stents produced contained up to 100 kBq of 133Xe and were implanted into the abdominal aortas of rabbits. Neointimal thickening was analyzed by histomorphometry for samples taken 4 weeks after stent implantation. The results indicate that the radioactive stents have a potential to suppress neointimal hyperplasia in rabbits. PMID- 10376329 TI - Detection limits for 90Sr, Pu, Am and Cm in soil and pasture vegetation shortly after a nuclear accident. AB - This work estimates the critical activity concentrations of 90Sr and the alpha emitting isotopes of Pu, Am and Cm in soil and pasture vegetation that would be required to exceed the action levels for foodstuffs recommended by the IAEA. The results show that the common detection limits for environmental analysis of these nuclides may be increased by orders of magnitude if the aim of the analysis is to determine whether or not the action levels will be exceeded. This information is useful in the development of more simple and rapid analytical methods to be used shortly after a nuclear accident. In addition to activity concentrations, the critical deposition densities on soil and grazing areas are estimated. Critical limits are also derived for 137Cs and 131I. PMID- 10376330 TI - Determination of 210Pb concentration in the air at ground-level by gamma-ray spectrometry. AB - 210Pb activity concentrations in the air of Palermo were determined by gamma-ray spectrometric analysis of 323 particulate samples collected in the period September 1995-December 1996. For each sample, the air filtered through a cellulose filter paper was 8600 m3 on average. The values of the daily activity concentration of 210Pb were ranging from 136 to 3390 microBq/m3. PMID- 10376331 TI - A contribution to the improvement of accuracy in the quantitation of THC. AB - The accuracy of a quantitative analysis is highly dependent on the quality of the reference standard. Although reference standards are more and more supplied with a certificate, laboratories may feel the need for additional acceptance testing. In general, confirmation of the purity of many solid reference substances can be obtained by a number of simple tests. However, verification of the true content of reference solutions may be complicated. A number of problems with the THC quantitation caused our interest for a verification method for the THC reference solution. The quantitation of THC is performed by gas chromatography with flame ionisation detector. The effective carbon number concept was used to predict GC/FID response factors. Equations and data are presented to calculate theoretical response ratios of cannabinoids. The experimental data for CBD and CBN were in excellent agreement with the theoretical ones. The paper shows that the response factors of CBD and/or CBN can be used for the calculation of the THC content of either reference solutions or cannabis samples. PMID- 10376332 TI - Analysis of etorphine in postmortem samples by HPLC with UV diode-array detection. AB - Etorphine is a synthetic narcotic analgesic usually used in veterinary medicine. It possesses an analgesic potency up to 1000 times greater than morphine and is therefore used in low doses, primarily for tranquilising large animals. For veterinary use, etorphine is usually available in its commercial formulation as Immobilon, when in combination with acepromazine or methotrimeprazine. Due to the potency of etorphine, only very low doses are required to produce adverse or fatal effects. This paper describes a method for detecting and quantifying etorphine using HPLC with UV diode array detection (HPLC-DAD) and demonstrates the advantage of the technique for the detection of Immobilon at low doses. In a forensic case involving Immobilon, the etorphine concentrations measured in postmortem femoral vein and heart blood specimens were 14.5 and 23.5 micrograms/l, respectively. No etorphine was detected in the urine. To our knowledge this is the first time postmortem etorphine concentrations have been reported. PMID- 10376333 TI - The interpretation of cocaine and benzoylecgonine concentrations in postmortem cases. AB - This study examined cocaine and benzoylecgonine concentrations in 100 consecutive deaths where either compound was identified in blood or urine specimens to determine whether any relationship between these concentrations and cause of death can be found. Forty-seven of the 100 cases were deaths attributed to cocaine, narcotic or combined cocaine and narcotic intoxication. There were 13 cases of cocaine intoxication where no psychoactive substance other than ethanol was detected. The mean cocaine concentration in these deaths was 908 ng/ml; three cases had cocaine concentrations greater than 2000 ng/ml, while the other ten cases had cocaine concentrations less than or equal to 700 ng/ml. The mean cocaine concentration in non-cocaine deaths where no psychoactive substance other than ethanol was detected was 146 ng/ml. This difference was not statistically significant. However, the average blood benzoylecgonine concentration in the 13 cocaine deaths was significantly higher than in the 19 non-cocaine deaths. A review of combined cocaine and narcotic deaths suggest that the narcotic is the main causative agent in these deaths. PMID- 10376334 TI - Philippine population database at nine microsatellite loci for forensic and paternity applications. AB - Allele frequency distributions for a Filipino population from the National Capital Region (NCR) were determined for eight STR loci: HUMF13A01, HUMFES/FPS, HUMvWA, HUMFOLP23, HUMD8S306, HUMCSFIPO, HUMTPOX and HUMTHO1; and a VNTR locus: D1S80. Statistical analysis showed that the nine loci showed no deviations from Hardy-Weinberg and linkage equilibrium rules. The average power of paternity exclusion for the nine loci is 0.9962 and the discriminating power is 1-2 x 10( 9). The data obtained from this study will be used as reference data for forensic DNA typing in the Philippines. PMID- 10376335 TI - Multivariate analysis ('forensiometrics')--a new tool in forensic medicine. Findings on the victim of sharp-force homicide can predict the inter-relationship with the perpetrator. AB - A multivariate 'forensiometric' technique (PLS-DA) was used to create a model to predict the relationship between homicide victim and perpetrator on a five-level scale: strangers, acquaintances, drinking companions, relatives and spouses. The model is based on findings on the victim and at the venue, and uses the following ten variables as 'predictors', listed in falling order of covariation with instances of close relationship between victim and perpetrator: Victim found in home, female victim, a single sharp injury, injuries to the upper extremity, superficial sharp injuries to the chest ('scratches'), ten or more sharp injuries, presence of defence injuries, total number of sharp injuries, male victim, and victim found outdoors. The model was based on 87 sharp-force homicides (the model-set) and was validated on another set of 43 sharp-force homicides (the test-set). In this test-set validation, 17 of 39 cases (44%) were correctly predicted according to the results of earlier combined forensic and police investigations, and a prediction one step away from the correct level was given in another 17 cases. These results were significantly better than would have been obtained by chance. PMID- 10376336 TI - Effect of methamphetamine abuse on the bone quality of the calcaneus. AB - BACKGROUND: Methamphetamine, a derivative of amphetamine, has been said to cause mainly mental dependency in humans, but only little is known about its physical dependency. Like other narcotics, it may have chronic effects on the human body, so, this study was planned to evaluate it by examining the bone quality of the skeletal system. METHOD: Among the convicts serving in Fuchu Prison, two groups of people were chosen according to their methamphetamine experience. The bone quality of the calcaneus of both abusers (n = 59, ages 41 +/- 11 years) and controls (n = 50, aged 45 +/- 13 years) was examined with an Achilles ultrasound bone densitometer. SOS (speed of sound) and BUA (broadband ultrasound attenuation), both of which were obtained from the measurement and are an indicator of the strength of bone, were compared between the two groups. RESULTS: The SOS of the abuser group was 1559 +/- 24 (mean +/- SD) m/s and this was significantly lower than that of the control group, 1570 +/- 27 (mean +/- SD) m/s (p = 0.017). The BUAs of the abuser group and the control group were 108 +/- 10 and 110 +/- 10 (mean +/- SD) dB/MHz, respectively, and there was no significant difference between them (p = 0.181). CONCLUSION: There was loss of SOS of the calcaneus in methamphetamine abusers. PMID- 10376337 TI - Repeated arson: data from criminal records. AB - The criminal careers of all arsonists convicted in former West-Germany between 1983 and 1985 who were found not to be responsible due to diminished responsibility for psychiatric reasons and a random sample (every third) of all criminally liable arsonists during the same period of time were followed up until 1994 by means of their trial records. Reappearance before the court for arson did not differ between the groups. When subjects are grouped by the additional occurrence of crimes other than arson, however, arsonists with diminished responsibility are the most problematic group: In comparison with all other subgroups, the arsonists who were partly responsible who did not commit any crimes other than arson showed the highest number of fire-setting incidents. Among the arsonists who committed other crimes as well, arsonists with diminished responsibility had the highest number of additional offences. PMID- 10376338 TI - Estimation of stress in child neglect from thymic involution. AB - It is difficult to evaluate the extent of stress in cases of suspected child abuse/neglect in a medico-legal autopsy. We have previously reported that stress due to abuse/neglect was found to have led to thymic involution. To elucidate the influence upon thymocytes differentiation, we compared the proportion of the thymocyte subpopulation in the thymus of a neglected child with one in an age matched control obtained from cardiac surgery. We found that the relative number of CD4+ CD8+ double positive (DP) thymocytes decreased in the neglected child. It was presumed that the selective decrease in the number of the immature DP thymocytes with CD3- to low bcl-2low caused the thymic involution in the neglected child. It was suggested that an alteration in the proportion of thymocytes subpopulation might be used as an index of stress in cases of child abuse/neglect. PMID- 10376339 TI - Acute fatal poisoning cases due to furathiocarb ingestion. AB - Seven cases involving acute fatalities due to ingestion of furathiocarb, a carbamate insecticide, are presented. Furathiocarb was detected in the gastric contents using thin layer chromatography (TLC) and gas chromatography/mass spectrophotometry (GC/MS), and quantified in the blood using a gas chromatograph equipped with a nitrogen-phosphorus detector (NPD). The fatal levels of furathiocarb in the blood ranged from 0.1 to 21.6 micrograms/ml. PMID- 10376340 TI - Fourteen shots for a suicide. AB - A 56-year-old man is discovered unconscious in a pool of blood in the kitchen of his house. According to findings, the man used a 22 long Rifle to fire 14 shots at his thorax with trajectories going from front to back, from right to left and on a nearly horizontal level. All the projectiles got into the left front side of his thorax and came out just under the back of his left armpit. One of them then got through his left arm and fractured his left humerus. According to the findings made on the premises and the position of the bloodstains, we think that man put his rifle against the wall, resting on a pipe. He fired, unloading two clips into his thorax. He had to handle the bolt of the rifle before each shot. To reload, he took the bullets which were on the nearby table on which blood marks can be seen. When reloading at a certain moment, he sat down in his armchair and when he wanted to stand up, he leaned on the armrests, on which blood marks can be seen. The last bullet was probably the one which went through his left arm, preventing him from keeping on shooting. His death, caused by a hypovolemical shock, was obviously very slow. PMID- 10376341 TI - Determination of orbifloxacin in rabbit plasma by high-performance liquid chromatography with fluorescence detection. AB - A simple and sensitive high-performance liquid chromatography method is developed for the determination of orbifloxacin (ORB) in rabbit plasma. Sample preparations are carried out by adding phosphate buffer (pH 7.4, 0.1 M) and extracting with trichloromethane. ORB and the internal standard, norfloxacin (NOR), are separated on a reversed-phase column using an aqueous phosphate buffer-acetonitrile (80:20, v/v) mobile phase. The concentrations of ORB and NOR eluting from the column with retention times of 2.16 and 3.09 min, respectively, are monitored by fluorescence detection at 338 (excitation) and 425 nm (emission). The method is shown to be linear from 4 to 1500 ng/mL (regression coefficient r2 = 0.999). The quantitation and detection limits are 4 and 9 ng/mL, respectively. Mean recovery is determined as 92% by the analysis of plasma standards containing 150, 750, and 1500 ng/mL. Inter- and intra-assay precisions were 4 and 3%, respectively. PMID- 10376342 TI - Rapid gas chromatographic analysis of drugs of forensic interest. AB - High-speed gas chromatographic (GC) screening for drugs of forensic relevance is performed using a commercial Flash GC instrument in which the chromatographic column is resistively heated at rates of up to 30 degrees C/s. Temperature programming conditions are varied in an experiment designed to evaluate trade offs between resolution and analysis time for a mixture of 19 drugs of abuse. All 19 components can be separated with excellent resolution in 90 s. Specific analytes can be analyzed even faster; for example, amphetamine analysis is completed in less than 20 s. Case studies of confiscated street drugs containing amphetamine, cocaine, and heroin are analyzed to evaluate the retention time repeatability. Ten replicate injections over a 2-day period for 3 different drug samples achieved retention time relative standard deviations in the range of 0.48 to 0.81%. PMID- 10376344 TI - A simple, convenient and direct method for assessing the purity of cobalamins. AB - A straightforward and simple, but powerful and direct, method is presented for both the detection and quantitation of cobalamin impurities in either commercial cobalamins or in metastable cobalamins (Cbls), such as RSCbls. The method is, quite simply, the use of the aromatic region of the 1H NMR of cobalamins; it is a method developed as an outgrowth of our work preparing metastable thiolatocobalamins (RSCbls) and is a method that proved necessary for characterizing those (and by inference other) cobalamins unstable to HPLC separation conditions (i.e., and, therefore, where the normally powerful HPLC method so commonly used in cobalamin chemistry fails). Despite considerable, prior, modern multidimensional NMR literature on cobalamins, the present method has not yet been indicated explicitly, nor has anyone reported previously the NMR data required to prove that the method works (i.e., the data for a series of cobalamins and their common impurities proving that they have different chemical shifts in the aromatic region of their 1H NMR when examined under identical NMR solvent, pH and other conditions). The direct NMR method is easy to perform, readily quantitated and applicable to species unstable to the HPLC conditions required to separate cobalamin impurities. The results have allowed quantitation of the 5-11% impurities in, for example, commercial HOCb1.HX, results which document that some commercially available cobalamins are not as pure as the manufacturers' claims. PMID- 10376343 TI - Synthesis, characterization and antitumor activity of copper(II) complex with nicotinamido-4-bis(2-chloroethyl)aminobenzaldimine. AB - Nicotinamido-4-bis(2-chloroethyl)aminobenzaldimine (NBAB) was synthesized and characterized by elemental analysis, IR and 1H NMR spectra. The complex of Cu(NBAB)2(NO3)2 was prepared in ethanol and characterized by elemental analysis, conductivity, cyclic voltammetry, IR, UV-Vis, fluorescence, CD and EPR spectra. The characteristic data suggest that the complex has an elongated octahedral structure, and NBAB behaves as bidentate in the keto form. The antitumor activities of NBAB and the complex against L1210 murine leukemia and K562 were investigated with both the MTT method and the colony formation test. The results in vitro indicate that antitumor activities of NBAB are superior to 2,2' chlorodiethylamine hydrochloride (nitrogen mustard) for L1210, and inferior to nitrogen mustard for K562, but the antitumor activities of the complex for both cell lines are superior to nitrogen mustard. PMID- 10376345 TI - La ions in precipitated hydroxyapatites. AB - Hydroxyapatites were synthesized by precipitation from an aqueous solution with La3+ (0-0.75%) and with carbonate (0-6.1%) at controlled pH 7.0. Uptake of La3+ was 90-95% complete. Relatively low Ca/P (1.54-1.63) ratios were attributed to nonstoichiometry. Carbonate in samples was identified by IR spectroscopy as B type carbonate. Lattice parameters of the hexagonal apatite structure were not affected by the La3+ content. Noncarbonated samples heated to 800 degrees C transform partially to beta-Ca3(PO4)2. Thermogravimetric analysis showed release of 0.4 mol adsorbed and 1 mol crystalline water up to 400 degrees C and decomposition of carbonate up to 900 degrees C in the samples. Luminescence data obtained for Gd-containing hydroxyapatites prove that Gd3+ ions are not incorporated in the precipitated hydroxyapatite. These findings suggest that, in the La-containing samples, La3+ is surface absorbed and not incorporated in hydroxyapatite. PMID- 10376346 TI - Comparison of the core size distribution in iron dextran complexes using Mossbauer spectroscopy and X-ray diffraction. AB - Mossbauer spectra of a series of iron dextran complexes in the intermediate temperature range where both sextet and doublet coexist may be used to obtain a qualitative description of the distribution of core sizes in these samples. Eight samples from five suppliers have been examined at 100 and 77 K. These differ markedly in the relative doublet contribution to the total spectral area and also in the hyperfine fields characterizing the sextets. The results indicate three distinct types of distribution. One sample from each type has also been examined at 4 K, where the doublet component has vanished and the hyperfine field distribution has become narrow and symmetric. These data are compared with estimates of average core diameters from X-ray line broadening. PMID- 10376347 TI - A cycloplatinated compound of p-isopropylbenzaldehyde thiosemicarbazone and its chloro-bridged derivative induce apoptosis in cis-DDP resistant cells which overexpress the H-ras oncogene. AB - cis-Diamminedichloroplatinum(II) (cis-DDP) is a widely used antitumour drug which produces important damage on the DNA inducing apoptosis in several cell lines. We have analyzed the cytotoxic activity of novel cyclometallated complexes of p isopropylbenzaldehyde thiosemicarbazone (p-is.TSCN) and their dimeric chloro bridged derivatives in murine keratinocytes transformed by the H-ras oncogene which are resistant to cis-DDP (Pam-ras cells). The data show that, in contrast with cis-DDP, the tetrameric cycloplatinated complex [Pt(p-is.TSCN)]4 and its dimeric chloro-bridged derivative [Pt(microCl)(p-is.TSCN)]2 have a good in vitro therapeutic index when comparing the cytotoxicity in Pam-ras cells to normal murine keratinocytes (Pam 212 cells) since they induce cell death in Pam-ras cells at drug concentrations significantly lower than those needed to kill Pam 212 cells. At equitoxic doses (IC90), both complexes produce characteristic features of apoptosis in Pam-ras cells together with a drastic decrease in levels of H-ras protein. These effects are not observed when the cells are treated with the IC90 of the cis-DDP drug nor the p-is.TSCN ligand. Altogether, these results suggest that the platinum compounds [Pt(p-is.TSCN)]4 and [Pt(microCl)(p is.TSCN)]2 might have potential as antitumour agents in view of their specific induction of apoptosis in cis-DDP resistant cells. PMID- 10376348 TI - Irreversible inactivation of purple acid phosphatase by hydrogen peroxide and ascorbate. AB - Treatment of the Cu(II)-Fe(III) derivative of pig allantoic fluid acid phosphatase with hydrogen peroxide caused irreversible inactivation of the enzyme and loss of half of the intensity of the visible absorption spectrum. Phosphate, a competitive inhibitor, protected against this inactivation, suggesting that it occurred as a result of a reaction at the active site. The native Fe(II)-Fe(III) enzyme was irreversibly inactivated by H2O2 to a much smaller extent than the Cu(II)-Fe(III) derivative, whereas the Zn(II)-Fe(III) derivative was stable to H2O2 treatment. The rates of inactivation of the Cu(II)-Fe(III) and Fe(II) Fe(III) enzymes in the presence of H2O2 were increased by addition of ascorbate. These results suggest involvement of a Fenton-type reaction, generating hydroxyl radicals which react with essential active site groups. Experiments carried out on the Fe(II)-Fe(III) enzyme showed that irreversible inactivation by H2O2 in the presence of ascorbate obeyed pseudo first-order kinetics. A plot of kobs for this reaction against H2O2 concentration (at saturating ascorbate) was hyperbolic, giving kobs(max) = 0.41 +/- 0.025 min-1 and S0.5(H2O2) = 1.16 +/- 0.18 mM. A kinetic scheme is presented to describe the irreversible inactivation, involving hydroxyl radical generation by reaction of H2O2 with Fe(II)-Fe(III) enzyme, reduction of the product Fe(III)-Fe(III) enzyme by ascorbate and reaction of hydroxyl radical with an essential group in the enzyme. PMID- 10376349 TI - Characterization of bis(isoorotato)diaquamagnesium(II) dihydrate: a potentially useful complex for magnesium supplementation. AB - The synthesis and crystal structure of a new Mg(II) complex of stoichiometry [Mg(isoor)2(H2O)2].2H2O, where isoor is the monoanion of 5-carboxyuracil (isoorotic acid), are reported. The structure, solved by single-crystal X-ray diffractometry, shows that it crystallizes in the triclinic space group P(-1) with Z = 1. The electronic, IR and Raman spectra of the complex are briefly discussed. Its thermal behavior was also investigated by means of thermal gravimetry (TG) and differential thermal analysis (DTA) measurements in an oxygen atmosphere. Dissolution studies support the usefulness of the compound for magnesium supplementation. PMID- 10376350 TI - Metal-ion speciation in blood plasma incorporating the bisphosphonate, 1-hydroxy 4-aminopropilydenediphosphonate (APD), in therapeutic radiopharmaceuticals. AB - In the quest for more effective pain palliation radiopharmaceuticals for metastatic bone cancer, this paper relates results obtained with 166Ho complexed to the bone-seeking bisphosphonate, 1-hydroxy-4-aminopropililydenediphosphonate (APD). APD is itself a bone cancer pain palliation agent and this work was therefore driven by the idea that the energetic beta-particle emitter, 166Ho, coupled with APD could afford a highly effective radiopharmaceutical in the treatment of bone cancer. Complex-formation constants for important blood plasma metal-ions were measured by potentiometry or polarography at 37 degrees C and I = 150 mmol dm-3. The latter technique was used for systems where precipitates formed at ligand-to-metal ratios appropriate for potentiometry. For trivalent lanthanides, neither electrochemical technique could be used. Animal tests showed that the 166Ho-APD complex was taken up primarily by the liver due to precipitation or colloid formation. PMID- 10376351 TI - Selective cone suppression by the L-M- and M-L-cone-opponent mechanisms in the luminance pathway. AB - We investigated how transient changes of background color influence the L- and M- (long- and middle-wavelength-sensitive-) cone signals in the luminance pathway. Motion identification thresholds were measured for a drifting sinusoidal grating (1 cycle/deg) modulated along different vector directions in L- and M-cone contrast space. The color of a central 4-deg-diameter region was briefly altered (500 ms) by incrementing or decrementing either L- or M-cone excitation. Incrementing L-cone and decrementing M-cone excitation produced a field that appeared reddish relative to the yellow surround. Likewise, incrementing M-cone and decrementing L-cone produced a field that appeared greenish. Motion identification thresholds were obtained on the yellow field following the brief color transitions. The results show that the threshold for the L-cone direction was selectively elevated by the background substitution of incrementing L-cone and decrementing M-cone excitation (shift toward reddish color). The same substitution, however, did not affect the threshold in the M-cone direction. Similarly, the threshold for the M-cone direction was selectively elevated by the background substitution of incrementing M-cone, decrementing L-cone excitation (shift toward greenish) without affecting the threshold in the L-cone direction. Experiments using the motion quadrature paradigm confirmed that these effects occur within the luminance mechanisms. These results indicate that the activation of L-on plus M-off signals suppresses the L-cone signal and that the activation of L-off plus M-on signals suppresses the M-cone signals in the luminance pathway. We propose a retinal model based on the experimental results. PMID- 10376352 TI - Reflectance and curvature of the inner limiting membrane at the foveola. AB - Light reflected specularly by the inner limiting membrane (ILM) provides information on the topography of the retinal surface. The ILM in the central part of the foveal pit acts as a concave mirror. Light reflected specularly by this mirror forms an image of the entrance pupil in front of the retina. In 15 normal subjects (ages 16-56 years) we have measured photometric and geometrical properties of this image to derive two characteristics of the ILM: its reflectance rho at the foveola and its radius of curvature r in the central part of the fovea. rho and r are found to decrease significantly with age (p = 0.0073 and p = 0.01, respectively). The equations of the regression lines are log10 rho = -4.234 - 0.0118 age and radius r = 1484 - 13.6 age, respectively (age in years, r in micrometers). PMID- 10376353 TI - Application of anterior pituitary cell culture in toxicological research. AB - High density plating procedure was used to evaluate the effect of atrazine on anterior pituitary cells of rats in monolayer culture. Collagenase-dispersed pituitary cells plated in suspension with medium-199 and 10% foetal calf serum attached quantitatively to plastic surfaces within 24 hours. Electron microscopy showed subpopulations of different cell types. After prolonged cultivation, most cells established small colonies with extensive contacts among them. Cell-to-cell formation of aggregates was significant and the colonies manifested morphological changes. The cells retained their enzymatic activity, converting testosterone into 5 alpha-dihydrotestosterone by enzyme 5 alpha-reductase. Immunohistochemical techniques facilitated differentiation of gonadotrophs producing follicle stimulating hormone (FSH) and luteinising hormone (LH). Atrazine in concentrations of 5 to 50 micrograms/ml of medium was associated with a significant reduction in the number of viable cells within 72 hours. The results suggest that the pituitary cell culture may prove useful in toxicological testing of various toxic compounds and reduce or replace in vivo animal experiments. PMID- 10376354 TI - Increase in lactate dehydrogenase isoenzyme-4 and splenocyte toxicity in methomyl treated rats. AB - The toxic effect of methomyl was studied in rats after a single or repeated oral administration. Rats treated with a single dose of methomyl (3, 5, or 7 mg/kg) showed significant increase (P < 0.05) in total lactate dehydrogenase (LDH) activity on day 1. The highest level of LDH activity was observed on day 3 in rats receiving 7 mg/kg of methomyl. The total LDH activity returned to normal on day 7 after dosing. Specific increases in LDH-3 and LDH-4 isoenzyme activities were observed. In rats treated with a single dose of 6 and 8 mg/kg of methomyl, spleen weight and splenocyte viability significantly dropped (P < 0.05) on days 1 and 3, respectively. Splenotoxicity was prevented by pretreatment with 60 mg/kg of N-acetylcysteine. The results suggest that the splenotoxic effect of methomyl is more likely directly related to oxidative cell injury than to cholinesterase inhibition. The significance of cytotoxic effects and the nature of cytotoxicity in relation to reactive oxidative damage deserve further investigation. PMID- 10376356 TI - Chemical composition of rainwater collected at two sampling sites in the city of Rijeka. AB - This study compares the chemical composition of rainwater samples collected at two sampling sites, the first situated in the Rijeka city centre and the second in a suburban site 120 m above the sea level. The rainwater samples were analysed for precipitation weighted average concentrations of hydrogen, sulphate, nitrate, chloride, ammonium, sodium, potassium, calcium, and magnesium. The results suggest that the local washout of the atmosphere enhanced the rainwater acidity in the city centre which also received significant marine contributions of sulphate, calcium, magnesium, and potassium content. Rainwater in the suburban site was affected by soil dust and/or fertilizers used in the nearby gardens, resulting in partial neutralization with rising of pH value. While the content of S-SO4 was practically equal at both sites, the quantities of N-NO3 and N-NH4 nearly doubled at the suburban site. PMID- 10376355 TI - Bacterial pollution of cutting fluids: a risk factor for occupational diseases. AB - An outbreak of occupational diseases involving 38 metal workers with local infections on the hands and the face (53%), bronchitis (29%), and bronchopneumonia (18%) urged an examination of 150 samples of emulsions which the workers used in processing metals. Among the isolated bacteria, the most frequent was Pseudomonas aeruginosa (48%), followed by Escherichia coli (44%), Proteus spp. (11%), Enterococcus faecalis (9%), Enterobacter aerogenes (7%), Citrobacter spp. (5%), Shigella spp., Staphylococcus aureus (3%), and the sulphite reducing Clostridia (1%). Moulds and yeasts were isolated from all tested samples. Most samples counted about 3 x 10(5)/ml of colony forming units. It was concluded that all emulsions and other substances which come in direct contact with the workers should be regularly monitored. Furthermore, preservatives should be added to prevent microbial infection. PMID- 10376357 TI - New technologies in diagnosing occupational asbestosis. AB - The paper considers the possibilities of applying telemedicine, a relatively new branch combining medicine and telecommunications, in diagnosing occupational asbestosis. Nowadays, telemedicine has been extensively used in an ever increasing number of areas such as dermatology, oncology, radiology, surgery, cardiology, and psychiatry. The paper gives an example of possible applications in Croatia. Telemedicine is expected to significantly reduce absenteeism and travel expenses incurred by various laboratory tests. Furthermore, the diagnostic procedure would be considerably quicker, thereby improving the overall health condition of the population living in the immediate vicinity of asbestos factories. Our research has been conducted in the southern Croatian town of Ploce with an asbestos plant, but possible application refers to places distant from major hospitals, such as rural areas and the islands. PMID- 10376358 TI - [Importance of assessment of visual function in nautical school students and naval engineers]. AB - Eighty-seven junior nautical school students, of which 46 undergraduate marine engineers and 41 undergraduate nautical officer were examined for visual functions as a part of screening procedure for nautical occupations. All students were male, aged between 17 and 18. Examination comprised visual acuity, visual depth, and colour vision. The visual functions were then quantified according to ergo-ophthalmologic criteria. Statistically, the visual functions were found to be within the normal limits. Only 5% of students bordered with visual impairments in acuity and depth. Five undergraduate marine engineers and one undergraduate nautical officer were found to be deuteranomalous. Deuteranomaly in the latter presented a serious impediment and the student was not eligible for the occupation. It turned out that he had not been examined before the enrollment. The enrollment in the nautical school should be conditioned by previous medical examination that would be performed in accordance with the Regulations for Nautical Personnel. The student's health should be monitored throughout the education and the examination should be repeated on entering the junior class. The existing Regulations for Nautical Personnel should be updated. PMID- 10376359 TI - [[Long-term bone marrow culture]. AB - Long-term bone marrow culture is an experimental in vitro model of hematopoiesis imitating conditions in vivo. It contains hematopoietic elements at various stages of differentiation as well as a supportive stromal microenvironment. Primitive hematopoietic stem cells of mesenchymal origin, the long-term culture initiating cells proliferate and differentiate into different cell types, giving rise to the adherent stromal layer and to various hematopoietic elements attached to it or floating freely in the supernatant medium. The stromal layer keeps the hematopoietic cells aggregated, helps their mitosis, differentiation and maturation by cell-to-cell contact, produces hematopoietic growth factors (cytokines), and forms the extracellular matrix required for cell attachment. Hematopoiesis occurs without exogenous growth factors. The appearance and development of the stroma, the proliferation and differentiation of hematopoietic progenitor cells, and the production of cytokines differ in long-term cultures of the normal and of pathologically altered bone marrow. Long-term bone marrow culture is applied in fundamental studies of normal and pathologically altered hematopoiesis, in pharmacological research, in the purging of residual leukemia cells from bone marrow autotransplants, and in the gene transfer. It is also suitable for testing carcinogenic and toxic chemicals causing hematopoietic damage through occupational or habitual exposure. PMID- 10376360 TI - [Information services in occupational medicine--comments]. AB - The recent social and public health policy changes have largely affected the functioning of occupational health units in two ways: the occupational physician has been reduced to an advisory function, while he has simultaneously been cut from the information sources such as community health centres, part of the Institute for Medical Research and Occupational Health, and scientific and professional publications. This has put the profession in an unfavourable position. The occupational physician requires fast, accurate, and comprehensive access to all relevant information. The author proposes establishing a single national information centre for occupational health that would collect, create, and distribute relevant and updated recommendations and information, legal regulations and updates, norms, and standards. PMID- 10376361 TI - Is information knowledge? Does it matter? PMID- 10376362 TI - Publish or perish. The debate from a global perspective. PMID- 10376363 TI - The hard choice facing Australian GPs. PMID- 10376365 TI - Growing pains. PMID- 10376366 TI - Insulin and type 2 diabetes. Last resort or rational management? AB - BACKGROUND: Recent evidence indicates that lower glucose levels in people with type 2 diabetes result in fewer complications. People with diabetes generally have sub-optimal glycaemic control. The natural progression of diabetes is characterised by increasing glucose levels requiring increasing therapy. OBJECTIVE: This article explores the possible role of therapeutic insulin in the management of type 2 diabetes. Arguments for earlier use of insulin, illustrative cases and common dilemmas faced when introducing insulin are examined. DISCUSSION: Findings from the United Kingdom Prospective Diabetes Study (UKPDS) are reviewed. It suggests that active and aggressive management of type 2 diabetes in general practice can have a role to play in reducing complications from diabetes. It now appears that insulin has a role earlier in the management of type 2 diabetes. PMID- 10376367 TI - Hormone replacement therapy. Indications, benefits and risk. AB - BACKGROUND: The debate surrounding oestrogen replacement in peri and postmenopausal women has been made overly controversial by the dissemination of misinformation and subjective evaluation of available data both by proponents and opponents of such treatment. OBJECTIVE: To review the indications, risks and benefits of oestrogen with and without progestin therapy in postmenopausal women. CONCLUSIONS: When prescribed judiciously, such that undesirable side effects are avoided, many women well benefit overall from the use of oestrogen in the postmenopausal years. Clearly many of these benefits are long term in addition to providing acute symptomatic relief. PMID- 10376368 TI - Syndromes of hyperandrogenism in women. AB - BACKGROUND: Although the actual incidence is not known, hyperandrogenism seems common among Australian women. Most women with syndromes of hyperandrogenism present with symptoms such as acne, hirsutism and androgenic alopecia, or menstrual irregularity. OBJECTIVES: The aim of this overview is: to put in perspective the various causes of hyperandrogenism in women, provide a guide to evaluation of affected women; and briefly review current management strategies. DISCUSSION: Hyperandrogenism in women is also associated with significant health problems including infertility, insulin resistance, gestational diabetes, type 2 diabetes mellitus and dyslipidaemia which may or may not be linked with obesity. PMID- 10376369 TI - Pituitary disease. AB - BACKGROUND: Pituitary disease is relatively uncommon, but because failure to recognise and manage it correctly has such severe repercussions, it has an importance out of proportion to its prevalence. OBJECTIVE: Pituitary disease may result in findings due to hormone excess or hormone deficiency. This article revisits the main clinical findings, investigations and treatment options for pituitary disease. DISCUSSION: The clinical presentation of pituitary disease varies from asymptomatic to severe features of endocrine disturbance. Establishing the diagnosis is not easy and requires a strong clinical suspicion supported by radiology and specific biochemical tests. Treatment is directed toward both the underlying pathology and the endocrine disturbances present. PMID- 10376370 TI - Suspicious skin lesions and their management. AB - OBJECTIVE: To describe the spectrum, diagnosis and management of non pigmented suspicious skin lesions in general practice. METHOD: General practitioners recorded all patients initiating consultations for suspicious skin lesions over 6 weeks in a tropical provincial city in Queensland as part of a study to determine the incidence of skin cancer. The spectrum of lesions and outcomes of the examinations and diagnoses are reported. Proportions of correct clinical diagnoses with 95% confidence intervals (CI) were calculated on excised lesions. Options in clinical management according to certainty of diagnosis were compared with chi-square statistics. RESULTS: Of the 81 eligible GPs 61 (75%) recorded detailed data on 1355 lesions. These included clinical diagnoses of 387 (28.6%) nonmelanoma skin cancers, 836 (61.7%) dysplastic lesions and 132 (9.7%) other benign lesions. 454 (33.5%) lesions were reported as excised or biopsied, 707 (52.2%) were treated without biopsy, 24 (1.8%) were referred to a specialist, 147 (10.9%) were monitored without treatment and 23 (1.6%) had no management specified. For lesions histologically confirmed by local pathologists as malignant (basal cell carcinoma or squamous cell carcinoma), the clinical diagnosis was correct in 69.1% of cases (95% CI 62.5-75.7%). The doctors reported managing 71.2% (95% CI, 65.6-76.7%) of clinically diagnosed BCC and 90.2% (95% CI, 85.6-94.9) of SCC by excision or biopsy. If more certain of the diagnosis of solar keratosis they were likely to treat without obtaining histology and if less certain they were likely to excise or biopsy (p = < 0.0001). CONCLUSION: GPs see a spectrum of skin lesions which are of concern to patients. A high proportion of these lesions are clinically benign and are not excised. If a BCC or SCC is suspected or diagnosis is uncertain most lesions are excised or biopsied. PMID- 10376371 TI - Diagnosing skin cancer in general practice. PMID- 10376372 TI - Asthma management in adults. Strategies to minimise morbidity. AB - BACKGROUND: Fundamental changes to the approach in asthma treatment over recent years have resulted in a reduction in asthma mortality in Australia. However, many factors still contribute to asthma morbidity. OBJECTIVE: This article identifies the preventable causes of difficult asthma and identifies those patients at greatest risk of mortality. Recent changes in the use of long acting beta agonists and inhaled corticosteroids are discussed. DISCUSSION: Morbidity due to asthma and quality of life for asthmatic patients can be improved by: identifying preventable causes of persistent asthma symptoms; reviewing inhaler technique; targeting high risk for more intensive monitoring; familiarity with the drugs used to treat asthma. PMID- 10376373 TI - Japanese encephalitis. PMID- 10376374 TI - Clinical audit activity. Prescribing for common conditions--hypertension. PMID- 10376375 TI - The speculum examination. PMID- 10376376 TI - Intermittent abdominal pain. PMID- 10376377 TI - Cases in travel medicine. After the banquet. PMID- 10376378 TI - Knowledge and intuition. PMID- 10376379 TI - Pretreatment symptom distress in women newly diagnosed with breast cancer. AB - Although the initial phase of illness is recognized as important in the overall process of adjustment after a diagnosis of breast cancer, little is known about pretreatment patterns of symptom distress. Seventy-four women ages 25 to 79 years and newly diagnosed with breast cancer were studied to determine physical, cognitive, and affective distress in the pretreatment period. Severity of distress was assessed about 11 days before primary surgery using the Symptom Distress Scale (SDS), Attentional Function Index (AFI), and Profile of Mood States (POMS). Higher levels of distress (SDS) were related to a triad of symptoms, insomnia, fatigue, and loss of concentration. Also, lowered effectiveness in cognitive function (AFI) and significant disturbances in mood state (POMS) were observed. Overall, a greater number of symptoms was associated with lowered cognitive function (r = -0.47; p < 0.01) and greater mood disturbance (r = 0.65; p < 0.01). Younger women younger than 55 years of age (n = 25) reported significantly (p = 0.02) greater overall symptom distress (SDS) than older women (n = 49). Interestingly, severity of distress did not differ in groups anticipating breast-conserving surgery (n = 35) instead of mastectomy (n = 39). The findings showed a discernible pattern of symptom distress before any treatment in women newly diagnosed with breast cancer, indicating a need for early intervention to promote the initial process of adjustment. PMID- 10376380 TI - Patient-related barriers to cancer pain management in a palliative care setting in Hong Kong. AB - This article reviews a study of pain management and its barriers in Hong Kong. Using an interview technique, several measures were used to understand the level of concern in patients about pain, the patients' hesitancy in reporting pain, use of analgesics, and adequacy of medication for pain. A total of nine barriers were identified, which include "addiction," "tolerance," "side effects," "physician distraction," "good patient," "fear of injection," "time interval," "fatalism," and "disease progression." Thirty-nine interviews were carried out. The interviewees were all cancer patients with pain in a palliative setting in Hong Kong. When the findings in Taiwan and the United States were compared, it was found that the cancer patients in Hong Kong had a higher level of concern toward the patient-related barriers. It was also found that the level of concern was generally higher in the group with hesitancy in reporting pain and using analgesics. Last of all, this project also identified the educational needs of patients and health care workers in Hong Kong. PMID- 10376381 TI - Classifying the empathic understanding of the nurse psychotherapist. AB - The purpose of this article is to identify types of verbal communication approaches to empathic understanding, and to consider the way of psychological support for cancer patients based on those types. These types were derived from the nurse psychotherapy process used with 46 Japanese cancer patients involved in the author's practice as a nurse psychotherapist. The psychotherapy process was interpreted by the phenomenologic approach. These types are as follows: Type A: Reflecting the perceived meaning of the verbalized experiencing of the patient. Type B: Anticipatively communicating the intuitively perceived meaning of the patient's experiencing, not verbalized, but now aware. Type C: Communicating the intuitively perceived meaning of the patient's experiencing, indifferent now, but usually aware. Type D: Facilitating the patient in focusing on her or his implicit felt sense. Type E: Communicating definitely the insight of the patient's implicit felt sense. These approaches indicated that when both a patient and a nurse psychotherapist have a deep level of empathic understanding, the patient could interpret personal meaning clearly and change the previous meaning. This experience helps the patient recover his or her own way of being, while helping the patient to live in the here and now. PMID- 10376382 TI - New agents in gastrointestinal malignancies: Part 1: Irinotecan in clinical practice. AB - In 1996 two chemotherapy agents were introduced by the U.S. Food and Drug Administration (FDA) with indications for the gastrointestinal malignancies for advanced colon and pancreatic cancers. The agents approved were irinotecan hydrochloride (CAMPTOSAR Injection, Pharmacia & Upjohn Company, Kalamazoo, MI; also investigated under the name CPT-11) for the second-line treatment of metastatic colorectal cancer, recurrent or relapsed, after 5-fluorouracil (5-FU) based therapy, and gemcitabine hydrochloride (GEMZAR for injection, Eli Lilly and Company, Indianapolis, IN; also referred to as dFdC) for first-line treatment of locally advanced and metastatic cancer of the pancreas. Irinotecan and gemcitabine, with demonstrated activity in colorectal and pancreatic cancer, respectively, are generally well tolerated and can be administered safely on an outpatient basis. Clinically relevant activity is documented for both single agents. Therapy-related side effects are manageable with appropriate monitoring and intervention, and reversible with dose modification or discontinuation. This article is one of a two-part series on new chemotherapeutic agents for gastrointestinal malignancies. The first in the series, this article addresses the agent irinotecan hydrochloride (CAMPTOSAR Injection). The second article, appearing in a subsequent issue, will review gemcitabine hydrochloride (Gemzar for Injection). Both articles review the current clinical use, safety profile, and key patient management guidelines for these new and novel cytotoxics. As clinical and investigational use of irinotecan and gemcitabine increases, the oncology nurse and other members of the health care team will need to anticipate potential treatment associated toxicities and be knowledgeable in their early identification and management. As patient advocates, oncology nurses play a key role in treatment outcome and related quality of life through expert patient education, symptom recognition, and intervention individualized to patient tolerance. This first article of the series addresses irinotecan, which in 1996 was approved for the second-line therapy of metastatic colorectal cancer, recurrent or elapsed, after 5-fluorouracil (5-FU). PMID- 10376383 TI - Long-term quality of life in patients after total gastrectomy. AB - This study used a questionnaire survey to evaluate the long-term quality of life (QOL) in 51 patients who underwent total gastrectomy. Activities of daily living (ADL) were good in 20, relatively good in 9, relatively poor in 13, and poor in 6 patients. The other 3 patients were treated on an inpatient basis. Of 38 patients who had been employed before surgery, 18 (47%) resumed working. Physically, body weight increased or showed no changes in 38 patients (74%), but decreased in the other 13 patients, of whom 3 showed a 15% or more decrease. Dumping symptoms developed in 13 patients (26%), 2 of whom had a severe condition. Clear decreases in physical strength and mental strength (spiritual energy) were reported by 10 and 8 patients, respectively. Comprehensive QOL was good in 20, slightly poor in 17, and poor in 14 patients. Quality of life was poor in 12 (41%) of the 29 patients with good ADL. The following were suggested as necessary for patients and their families: sufficient preoperative explanation about pathophysiology and nutritional management after total gastrectomy; execution of a continued patient education program until after discharge; and explanations in specific terms about cooking methods, nutritional management, exercise therapy, periodic medical checks, and patients' associations using pamphlets and food models to describe daily living of the patients after discharge. PMID- 10376384 TI - American Cancer Society's Program of Professorships in Oncology Nursing: survey of activities of the recipients. PMID- 10376385 TI - Nurses' perspectives on unconventional therapies. AB - Unconventional therapies have become increasingly popular with health care consumers in recent years. As patients seek information and attempt to make decisions about unconventional therapies, they often turn to nurses, asking the nurse's opinion about certain therapies. The nurse's attitudes and beliefs about unconventional therapies quite likely will influence the response to the patient's inquiries. This article represents the findings of interviews with 20 nurses regarding their perspectives on unconventional therapies. Without exception, all nurses who were interviewed emphasized that information regarding unconventional therapies needs to be available readily for both patients and health care professionals. Other themes identified in the interviews included the following: Various people use unconventional therapies; people seek unconventional therapies for a variety of reasons; communication about unconventional therapies needs to be open, and a place should be found for unconventional therapies. The interviewees saw a clearly defined role for nurses regarding unconventional therapies. PMID- 10376386 TI - Factors associated with acceptance or rejection of recommendation for chemotherapy in a community cancer center. AB - Socioeconomic, attitudinal, and psychological factors associated with acceptance or refusal of recommendations for chemotherapy were investigated in 64 consecutive patients with solid tumors or lymphoma who agreed to participate in this study. Patients filled out the Brief Symptom Inventory (BSI) and a questionnaire that investigated selected factors. Patients also were asked if they believed in, used, had used, or planned to use alternative-complementary treatments for their cancer. Eight patients refused chemotherapy against the advice of their oncologist. Frequency, percentage, mean, standard deviation, and range was coded, and differences between the groups of those who accepted and those who rejected recommendations for chemotherapy were analyzed by chi-square, using unpaired t test and the Wilcoxon two-sample test. There was a significant increase in anxiety in the total study population as compared with patients who did not have cancer. In addition, all the BSI scores except those for anxiety were higher in patients who refused chemotherapy, and the difference was statistically significant. PMID- 10376387 TI - [Health care needs and their assessment. Part 2. The need for health and health care]. AB - Health needs assessment is primarily concerned with the benefits that can be expected from the provision of different kinds of health services. These are closely related to the levels of health risks and illness in the population and the expected effectiveness of the service in reducing health risks and the consequences of illness as indicated by results of evaluative research or by clinical consensus. Levels of disease and health risks in a population are commonly referred to as needs for health, and the levels of expected benefit are referred to as the needs for services or care. PMID- 10376388 TI - [Genetic predisposition in multiple metabolic syndrome. Part 2. Candidate genes in type II diabetes mellitus]. AB - The author presents a review on candidate genes of proteins involved in the metabolism of glucose, lipids and other metabolites (glucose carriers, insulin receptors, proinsulin, glucokinase, amyline, glycogen synthase). One of the main causes of enhanced atherogenesis in patients with type II diabetes (NIDDM) are marked genetically conditioned deviations of the lipid, lipoprotein and apolipoprotein metabolism. In the metabolic dyshomeostasis of multiple metabolic syndrome participate in the process of atherogenesis also: isoforms of apolipoprotein E4, isoforms of apolipoprotein A-IV-1/1, hyperuricaemia, raised levels of the plasminogen activator inhibitor 1 (PAI-1), hyperfibrinogenaemia, hyperhomocysteinaemia and other metabolites (cytokines, endothelin etc.). Patients with a greated genetic sensitivity manifest diabetes sooner and more intensely and die at a younger age in particular from cardiovascular disease, but also on account of a higher incidence of tumours diseases. PMID- 10376389 TI - [Molecular genetic analysis of Alzheimer's dementia in the Czech population. The APP-717 mutation in the gene for amyloid protein precursor]. AB - BACKGROUND: Although locus mutations in the gene for the amyloid precursor protein were already described in patients with Alzheimer's disease, there still are some patients where this mutation was not found and no link was found with other possible genetic loci on chromosomes 14 and 19. Therefore a group of subjects with Alzheimer's disease was subjected to tests for the presence of a mutation in the APP gene (in position 717). METHODS AND RESULTS: In a selected group of subjects with Alzheimer's disease (AD) in the gene for amyloid precursor protein in position 717 mutations of its transmembraneous region are found. The authors analyzed the genome DNA of cerebral tissue of Czech subjects for the presence of this mutation by means of the polymerase chain reaction with subsequent verification by sequencing analysis. In every subject genetic analyses from cerebral areas of the frontal lobe, temporal lobe, parietal lobe and hippocampus were performed. The methods used were the polymerase chain reaction (PCR) and sequencing. From the total number of 18 subjects with confirmed Alzheimer's disease and six non-related subjects without histopathological signs of Alzheimer's disease after the age of 90 years, three sequencing changes were found in position 717 of exon 17 of the transmembranous region of the precursor of beta-4 amyloid glycoprotein. In the first case it was substitution of thymin for adenine in codon 717, in the second case substitution of cytosine for thymine, in the third case a sporadic mutation of guanine for thymine in codon 717 was found. CONCLUSIONS: It was revealed that codon 717 could be a so-called hot spot site preferred for the preferential development of mutations in codon 717 in the gene for the amyloid precursor protein (APP). PMID- 10376390 TI - [Budesonide (Turbuhaler) at a dosage of 400 micrograms per day is at least as effective as a double dose of beclomethasone (MDI) in patients with mild to moderately severe bronchial asthma]. AB - BACKGROUND: Recent investigations revealed that in patients with bronchial asthma the same anti-inflammatory effect is achieved by inhalation of half the dose of budesonide by a Turbuhaler (i.e. by using corticosteroid in powder form) as by a full dose of beclomethasone driven into the lungs by compressed chlorofluorocarbons (i.e. MDI = pressure dosage inhalator). The objective was to assess whether there is a difference between 12-week treatment with budesonide Turbuhaler in a smaller dose of 400 micrograms/day and treatment with beclomethasone dipropionate MDI 800 micrograms/day. METHODS AND RESULTS: After an initial two-week period of the 227 patients with mild or medium severe asthma who had not taken corticosteroids for three months, into the budesonide Turbuhaler group 94 patients were included and into the beclomethasone MDI group 99 patients. Characteristics: group treated with budesonide (46 men, 48 women, mean age 38 years, FEV1 78% of appropriate values). Group treated with beclomethasone (51 men, 48 women, mean age 39 years, FEV1 81.5% of appropriate values). Morning and evening values of the peak expiration rate (PEF) increased significantly after budesonide treatment 2 x 200 micrograms/day) as compared with beclomethasone treatment (2 x 400 micrograms/day). Differences of morning PEF between budesonide and beclomethasone: 47:28 l/min, p < 0.05, differences of evening PEF: 32:10 l/min., p < 0.027. The number of dyspnoic attacks declined after both types of treatment, as well as the amount of inhaled bronchodilatating substances (terbutalin Turbuhaler). The differences between drugs were however not statistically significant. CONCLUSIONS: Budesonide Turbuhaler, 400 micrograms/day when administered to patients with bronchial asthma was at least as effective as beclomethasone MDI, 800 micrograms/day. The increase of morning and evening PEF values was after budesonide significantly higher than after beclomethasone. PMID- 10376391 TI - [Talc--a standard drug in the treatment of malignant pleural effusion]. AB - Currently, talc is the most effective and widely-used agent for chemical pleurodesis in the treatment of both malignant pleural effusion and pneumothorax. Its popularity has been growing due to the low incidence of side effects, low cost and higher success rate in comparison with other agents (tetracyclines, bleomycin, Corynebacterium parvum). The guidelines for talc pleurodesis are summarized and the history of the Corynebacterium parvum as an agent for chemical pleurodesis is mentioned. Its production in the Czech Republic has been halted due to the increased interest in the talc as a chemical sclerosant. PMID- 10376392 TI - [Use of the publications and documents of the World Health Organization]. AB - For 50 years the development of reliable information has been one of the functions of WHO. Much of this information is made available through an extensive programme of publications. The National Medical Library. Prague as the WHO Documentation Centre acts as a national support to the promotion, distribution and reproduction of WHO documentation. PMID- 10376393 TI - [Development of European medicine to the time of Vesalius (16th century)]. AB - Mythical Greece, Egyptian Alexandria and ancient Rome were the cradle of ancient medicine where the beginnings of anatomy and physiology were combined with nature philosophy of great personalities such as Hippocrates and Galen. A modern contribution i.e. medical chemistry, to European medicine was made by Arab physicians Rhazes and Avicena. Only when clerical dogmatism was overcome, after the discovery of the New world and the development of universities the road to scientific progress ws opened on which started the great surgeon and physician Andreas Vesalius. PMID- 10376394 TI - [Discussion of the article by P. Dlouhy, M. Andel: Trans fatty acids in the diet their possible health risks. PMID- 10376395 TI - [Priapism]. AB - Priapism is prolonged, and usually painful, erection not associated with sexual desire. It is a relatively rare acute urological disease where treatment must be started within 6 hours after its development, as after a longer time interval due to ischaemia irreversible fibrotic changes of the cavernous tissue of the penis develop which lead to permanent erectile dysfunction. There may be either low flow priapism (inadequate outflow of blood from the cavernous tissue) or more rarely high flow priapism (excessive inflow of blood). Priapism is classified with regard to its aetiology into primary (cause unknown) or secondary. The causes of secondary priapism are most frequently overdosage of vasodilatating agents during intracavernous injection treatment of erectile dysfunction (specially papaverine), tumours (obstruction of the efferent veins or direct infiltration of the corpora cavernosa)--in particular carcinoma of the urinary bladder, prostate and rectum. Priapism is frequently due to injuries of the prostate and straddle injuries. 5% men with sickle-cell anaemia suffer from an attack of priapism. Treatment of priapism differs, depending on the type, and should be entrusted to an experienced urologist in an in-patient department. PMID- 10376396 TI - [Genetic predisposition in multiple metabolic syndrome. Part 3. Metabolism of lipids, lipoproteins and apolipoproteins]. AB - The author discusses metabolic processes during exogenous and endogenous lipid transport and deviations in the metabolism of lipids, lipoproteins and apolipoproteins in multiple metabolic syndrome and in so-called diabetic dyslipidaemia. Specific phenotypic manifestations of diabetic dyslipidaemia include hypertriacylglycerolaemia, hypercholesterolaemia, elevated plasma levels of LDL-cholesterol and apolipoprotein B and reduced levels of HDL-cholesterol and apolipoprotein B and reduced levels of HDL-cholesterol and apolipoprotein A-I. Other recent findings relating to this syndrome include evidence of elevated concentrations of small and dense LDL micelles (< 25 nm), so-called LDL phenotype B, which are easily modified (e.g. by oxidation, glycation etc.), and subsequent uptake by "scavenger" receptors into macrophages which after filling become foam cells and penetrate into the vascular wall. Elevated levels of small and dense LDL micelles, the accelerating process of atherogenesis, were proved in all multiple metabolic syndrome carriers. The atherogenic lipoprotein phenotype hastens markedly atherogenesis and subsequent manifestation of cardiovascular diseases. PMID- 10376397 TI - [Type I and III procollagen in patients with abdominal aorta aneurysms]. AB - BACKGROUND: Collagen and elastin are the basic building stones of the aortal wall. During the process of development of an aneurysm of the abdominal aorta (AAA) degradation and remodelling of elastin and collagen in the intercellular matrix of its wall occurs. The two main types of collagen in the aorta are collagens type I and III. During type I collagen synthesis the carboxyterminal propeptide of procollagen (PICP) is released and during synthesis and degradation of collagen type III the aminoterminal propeptide collagen III (PIIINP). The objective of the present work was to assess to what extent the two factors can be used to follow up metabolic processes in the AAA wal and in plasma in relation to the extent and symptomatology of AAA. METHODS AND RESULTS: Samples of venous blood and the AAA wall were examined using radioimmunoanalytical methods. PIIINP levels in venous blood were significantly higher (wall p < 0.01) in patients with AAA (n = 78) as compared with the control group (n = 15). The authors did not reveal a statistically significant difference between the levels of the two factors in the blood of patients with AAA of different extent and symptomatology. The PIIINP concentration in the AAA wal correlated significantly with its extent and symptomatology (wall p < 0.01). CONCLUSIONS: Evidence was provided of an enhanced metabolism of collagen type III in the AAA wal with predominant degradation in growing and symptomatic AAA. For complete evaluation of the importance of PICP and PIIINP plasma levels it will be necessary to follow up their dynamics in subjects with growing, small (< 5 cm) AAA. PMID- 10376398 TI - [Warning signs preceding the development of septic shock in patients with neutropenia treated for hematologic malignancies]. AB - BACKGROUND: In patients with haematological malignities infectious complications take a very rapid course, and in particular during the period of neutropenia, they are not necessarily manifested by a clear symptomatology. Frequently they may be manifested only by an elevated temperature and general deterioration of the condition. The onset of shock then can be rapid and surprising. The objective of the work was to identify clinical and laboratory signs warning against the possible development of septic shock. METHODS AND RESULTS: The investigation comprised a total of 38 patients hospitalised due to infectious complications at the intensive care unit because of general deterioration of the condition. 18 developed septic shock (group S), the remaining 20 (group N) achieved during hospitalisation at the ICU a stabilised condition. In both groups the laboratory values and clinical condition were followed up for 2-3 days prior to deterioration of the condition. Risk factors for development of septic shock in the group of investigated patients were: unusual weakness, heart rate above 95/min. beyond the temperature peak, hypoalbuminaemia, mucositis, hypokalaemia, presence of a central venous catheter and administration of parenteral nutrition. PMID- 10376399 TI - [Etiopathogenesis and therapy of dermatophytosis of the nails]. AB - The importance of mycotic infections of the skin and its adnexa has a rising trend. The causal agents are not only dermatophytes and candidae but to an increasing extent opportune fungal microorganisms. Most frequently these diseases develop during intense immunosuppressive treatment, e.g. in oncological, oncohaematological diseases and in developed AIDS syndrome. In addition to the reduced defence capacities of the organism favourable conditions are created e.g. by antibiotic treatment, treatment with female hormones, unsuitable clothing, obesity, a certain part is played also by occupational predisposition. Treatment of onychomycoses is complicated and difficult. Before griseofulvin was available there was only surgical ablation and treatment with keratolytics. At present a number of substances with marked fungistatic effects is available. The author mentions: griseofulvin, ketoconazole, fluconazole, itraconazole (azole derivatives), allylamines (terbinafine-Lamisil). He discusses the mode of administration, dosage and undesirable effects. The recent antimycotics are ciclopiroxolamines (Batrafen) and amorolfin (Loceryl) in the form of lac. PMID- 10376400 TI - [Psychological factors, panic disorders and heart diseases]. PMID- 10376401 TI - [Insulin and insulin therapy]. AB - The author summarizes basic findings on the formation of insulin, its secretion and bond to insulin receptors and other phenomena at the post-receptor level. He summarizes basic indications for insulin treatment also beyond the sphere of diabetology. In a separate part the author summarizes briefly the history since the discovery of insulin up to the present time with modern therapeutic procedures (insulin injections, insulin pumps). The greatest part of the paper is devoted to contemporary insulin preparations and their use in therapy. A separate section is devoted also to insulin absorption and ways to influence its action. Attention is drawn to some differences in insulin absorption in relation to the site of injection, insulin concentration, body temperature and other factors which can be influenced. The objective of the paper is an overview of insulin and its use in therapy since to discovery of insulin to the present time. PMID- 10376402 TI - Treatment of children and adolescents with non-Hodgkin's lymphoma (results based on the NHL Berlin-Frankfurt-Munster 90 protocols). AB - BACKGROUND: To determine the feasibility and results of treating children with non-Hodgkin's lymphomas (NHL) according to very intensive protocols based on the German Berlin Frankfurt Munster NHL 90 study. METHODS AND RESULTS: From 1991 until 1995 eighty two patients less than 18 years of age with NHL were admitted to our department. Sixty three of them were eligible for the study. The entire group consisted of 43 males and 20 females (ratio 2.1:1). Median age was 10 2/12 years. Eleven had stage I disease, 4 stage II, 29 stage III and 19 stage IV disease. Histologies represented were: large cell lymphoma 22, lymphoblastic lymphoma 19, and Burkitt lymphoma 10 patients. In 12 cases the immunophenotype was not further classified as to B-cell or T-cell subtype. Patients were stratified into the therapy groups "B" or "non B" according to histopathology, clinical stage and LDH level. Therapy for the B group consisted of 2, 4 or 6 courses of intensive 5 day pulses of 6 drugs. Patients in the non B group received the protocol for acute lymphoblastic leukemia including reinduction and CNS irradiation for advanced stages. At a median follow-up of 35 months the probability of event free survival (pEFS) at 5 years 70% and overall survival 73% for entire group. For therapy group B pEFS was 76%. The non B therapy group had a pEFS 60% (p = 0.22). There was a significantly better outcome for children classified as stage I and II. There was no statistical difference between stage III and IV. Treatment results were comparable between NHL subtypes, except for large cell lymphomas, which did significantly better (pEFS 90%). CONCLUSIONS: The use of protocols based on BFM 90 study in the Czech Republic was feasible. The pEFS are approximately 10% lower than the German study but comparable to some other studies. Outcome for large cell lymphomas was excellent. Reduction of treatment related complication and mortality rate as well as more precise classification are required. PMID- 10376404 TI - [The first death from tertian malaria in the Czech Republic]. AB - The authors describe a fatal case of tertian malaria (Plasmodium vivax) imported to the Czech Republic from India. Severe clinical symptoms leading to shock and multi-organ failure are more frequent in tropical malaria (P. falciparum). Despite the use oc continuous veno-venous haemodiafiltration HLA-DR decline and death occurred. The effect of the administered antimalarias drugs was good. PMID- 10376403 TI - [Detection and significance of chromosome abnormalities in patients with malignant non-Hodgkin's lymphoma using the polymerase chain reaction]. AB - BACKGROUND: Morphology and immunological marker analysis are insufficient to detect neoplastic population in some cases (15%) of non-Hodgkin's lymphomas (B NHL). Aim of the study was to detect malignancy at molecular level using polymerase chain reaction. METHODS AND RESULTS: We examined a diverse set of B NHL (90 patients--48 men, range 18-76 years, mean age 53 years and 42 women, range 20-86 years, mean age 54 years) to detect immunoglobulin heavy chain (IgH) rearrangement. 32 patients with centroblastic-centrocytic lymphomas (12 men, range 45-64 years, mean age 52 years and 20 women, range 29-85 years, mean age 53 years) were also studied for translocation (14,18). DNA was isolated from lymphatic nodes, bone marrows and peripheral blood. Translocation (14,18) was founded in 38% lymphatic nodes, 36% bone marrows and in 50% of peripheral blood. The detection rate of IgH PCR varied according to the morphologic type of the analyzed lymphoma specimen. A high detection rate (100%) was observed in low grade lymphoma, while in high-grade lymphoma was in 62%. In bone marrows samples from follicular lymphomas, IgH PCR positivity was observed in 50% cases without leukaemic blood picture and in 64% cases with lymphoma cells in peripheral blood picture. In peripheral blood with bone marrow infiltration, but without the presence of lymphoma cells (morphological assessment) we observed 71% IgH PCR positive samples. In case, when bone marrow and peripheral blood were morphologic negative, we identified 64% positive cases. Using t(14,18) and IgH PCR we detected neoplastic population in 81% follicular lymphomas. CONCLUSIONS: IgH PCR and t(14.18) PCR are convenient additional technology for detection of neoplastic lymphocytes in B-NHL, particularly when morphology and immunological marker analysis are insufficient. PMID- 10376405 TI - [Evaluation of drug utilization from positive lists]. AB - BACKGROUND: The introduction of positive drug lists (positive lists) is part of the regulating mechanisms which try to reduce the increasing expenditure on drugs. An essential part of these provisions is at present an analysis of the trend of drug consumption. The objective of the work was to introduce and evaluate the method DU 90% which in analyses of drug consumption uses a 90% ratio of preparations of their total consumption. METHODS AND RESULTS: The analysis was focused on prescriptions in medical clinics of the General Faculty Hospital Classification of drugs according to anatomical, therapeutic and chemical groups and the use of defined daily doses made it possible to calculate DU 90% and to prepare a consecutive list of drugs according to costs in different departments. The number of drugs with a defined daily dose was within the range of 280-316. In the interval DU 90% were 116-142 drugs. In all departments there was a practically identical ratio of drugs which were part of the positive list. The greatest expenditure were anti-infectious drugs for systemic use and drugs with an effect an blood and haematopoietic organs. CONCLUSIONS: The advantages of the method DU 90% is its easy application, low cost and possibility to identify controversial lields. This facilitates further qualitative and quantitative evaluation of the standard of pharmacotherapy. PMID- 10376406 TI - [Selenium and the organism]. AB - Selenium is an essential trace element for animals. It is biologically active as selenocysteine in the active centre of selenoproteins with enzymatic functions. Incorporation of selenocysteine occurs on the basis of genetic expression and selenium is the only trace element under direct genetic control. Selenocysteine can be considered the 21st amino acid with regard to its biosynthesis and incorporation into proteins. At least two types of selenoproteins are necessary for each animal cell, the first from the family of GSH-peroxidases and the second from the family of deiodinases. GSH-peroxidases are the most powerful antioxidant enzymes, which defend the cell and whole organism against oxidative damage and thus from oxidative diseases and disorders such as cardiovascular diseases, malignancies, bacterial or viral diseases, muscle dystrophy, arthropathy, arterial plaques, and others. GSH-Px have many other regulatory functions such as regulation of biosynthesis of prostaglandins, prostacycline, leukotrienes, and thromboxans. Deiodinases regulate the metabolism of biologically active triiodothyronine and thus thyroid hormone regulation of the whole organism. Selenoproteins act against cancerogenic effects of some organic molecules and bind heavy metals. Tissue-specific selenoproteins without a known biological function have been detected in some specialised tissues with a high priority for selenium. One of the regulators of selenoprotein synthesis is the selenium status of the organism. Its state an intake may be assessed by analyses of selenium indexes. The most often used indexes are serum selenium and urinary selenium. On the basis of its analyses in six regions of the Czech Republic, severe selenium deficiency has been found in inhabitants of this country, which is even profound for more distressed groups like growing children, pregnant and lactating women, and the elderly. PMID- 10376407 TI - [Renal anemia and the effect of long-term dialysis]. AB - Renal anaemia causes in patients with chronic renal failure numerous serious problems which can be favourably influenced by improvement of the anaemia. There is a number of known factors which cause deterioration of anaemia and make its treatment more difficult. For a long time it was not clear that these negatively acting factors included also insufficiently effective dialysis treatment. The authors of the submitted paper evaluate the relationship between anaemia and the effectiveness of dialysis based on new data reported in the literature and their own results. From this evaluation ensues that inadequate haemodialysis, assessed from the percentage reduction of urea in blood, is associated with a reduced response to recombinant human erythropoietin which is the basic remedy of renal anaemia. If the inadequate intensity of haemodialysis is increased, anaemia improves substantially. In patients on continuous ambulatory peritoneal dialysis (CAPD) there is a direct relationship between the effectiveness of blood purification expressed by the index KT/Vurea, i.e. the indicator of urea elimination, and the severity of anaemia. In patients treated by CAPD there is a significant association between the haematocrit and KT/Vurea supplied by the peritoneum as well as the kidneys. KT/Vurea supplied by the patient's own kidneys is from the aspect of anaemia more significant. Some facts regarding the relationship between anaemia and the effectiveness of dialysis treatment remain obscure so far. This however does not influence the fact that based on data available at present, effective dialysis must be included among basic prerequisites of effective treatment of renal anaemia in dialyzed patients. PMID- 10376408 TI - [Age and changes in dietary habits affect hyperlipoproteinemia after kidney transplantation]. AB - BACKGROUND: Atherosclerosis of the blood vessels is the most frequent cause of morbidity and mortality of patients after transplantation of the kidneys with a long-term function of the graft. It is assumed that the most significant risk factor for its development is secondary hyperlipoproteinaemia (HLP). In the development of HLP a number of factors may participate (chronic renal insufficiency, proteinuria, immunosuppressive treatment, diet, increment of body weight, age, genetic factors). The objective of the investigation was to follow changes of the lipid spectrum after renal transplantation and evaluate the impact of different factors which participate in these changes. METHODS AND RESULTS: The authors investigated in a retrospective metabolic study for a period of 18 months a total of 348 patients after the first cadaverous kidney transplantation. They compared the findings in 34 patients (group I) who had throughout the investigation period a total cholesterol of < or = 5.2 and triacylglycerols < or = 2.3 and group II (314 patients) who had elevated values of these factors. The mean values of different parameters differed highly significantly (group I vs. group II): cholesterol 4.6 +/- 0.4 vs 6.8 +/- 1.5 (p < 0.001), triacylglycerols 1.8 +/- 0.8 vs 3.6 +/- 1.6 (p < 0.001), LDL-cholesterol 2.6 +/- 0.6 vs 4.0 +/- 1.1 (p < 0.001), (all in mmol/l) and HDL/total cholesterol 0.28 +/- 0.07 vs 0.20 +/- 0.09 (p < 0.01). The authors did not detect a difference in the incidence of isoforms of apo E. There were no differences in the mean cyclosporin A levels: 394 +/- 114 vs. 489 +/- 202 (ng/ml) and renal function CCr 1.0 +/- 0.6 vs 0.9 +/- 0.3 (ml/s). In group I there was a significantly higher intake of energy 150 +/- 20 vs. 125 +/- 25 (kJ), of fat 1.6 +/- 0.3 vs. 1.0 +/- 0.2 (g/kg) and disaccharides (by 50%). With this corresponded also a significantly higher increment of BMI 27.4 +/- 4.6 vs. 23.8 +/- 3.3 (p < 0.01). Patients in group II were also significantly older (44.5 +/- 14.1 vs 49.4 +/- 12.5, p < 0.01). CONCLUSIONS: It is assumed that one of the main causes of the development of HLP after transplantation are poor dietary habits of the patients associated with an excessive intake of energy, fats and disaccharides and an increase of body weight. The patient's age is significantly higher. Standard lower doses of immunosuppressive drugs have obviously only a supportive effect. PMID- 10376409 TI - [Immunohistologic determination of extracellular matrix components in granuloma annulare]. AB - BACKGROUND: The histological diagnosis of granuloma annulare is based on assessment of a palisade-like granuloma and necrobiosis. The authors attempt to make a more accurate diagnosis of the disease based on immunohistological assessment of components of the extracellular matrix. METHODS AND RESULTS: In a group of 15 patients with granuloma annulare the authors assessed, using the immunoperoxidase method, collagens type I, III and V and fibronectin. An increased amount of collagens type III and V was found within the palisade-like granuloma and its neighbourhood resp. In the area of the necrobiosis the findings of the mentioned collagens depended on the degree of necrobiotic changes (12 of 15 patients). An increased occurrence of collagen type I was only indicated in the vicinity of granulomas or within the palisade-like granuloma in 7 of 15 patients. Larger amounts of fibronectin were found in the area of the necrobiosis and in the area of the granulomatous palisade-like infiltrate resp. (12 of 15 patients). CONCLUSIONS: In the author's opinion the higher incidence of collagens types III and V in the neighbourhood of the palisade-like granulomatous infiltrate and its size resp, as well as the finding of fibronectin in the area of necrobiosis and the granulomatous infiltrate justify the use of immunohistological assessment of the above constituents of the extracellular matrix as a supplementary examination in the diagnosis of the above disease. PMID- 10376410 TI - [Mushroom poisoning by Cortinarius orellanus]. AB - The authors present the case-records of three patients who became intoxicated with the mushroom Cortinarius orellanus. This mushroom is very rare in this country and is not well known. The toxin orellanin is solely nephrotoxic and renal affection can lead to acute renal failure. A specific feature of this intoxication is the symptom-free period from 2 to 21 days, gastrointestinal complaints associated with back pain. The diagnosis can be established from a mycological analysis or by estimation of toxin in serum or tissue obtained by renal biopsy. The basis of treatment is disposal of the toxin by extracorporeal elimination methods: haemodialysis and haemoperfusion. As with the length of the interval between intoxication and onset of treatment the probability of irreversible renal affection increases, the medical community should take into consideration possible intoxication with this mushroom in the differential diagnosis of acute renal failure. PMID- 10376411 TI - [The Masaryk Homes and the Charles University Medical School in Prague]. PMID- 10376412 TI - [The Slovak Pharmaceutical Society between the 5th and 6th congresses]. PMID- 10376413 TI - [Antioxidants of biological origin]. AB - Diseases due to free radical are the subject of intensive study. The search aims to find substances preventing the origin of free radicals, substances capable of rendering the developed free radicals harmless, or substances capable of removing the damage of molecules which has already taken place. The present paper briefly surveys the metabolites of biogenic origin, mainly from higher plants, which show in vitro antioxidant activity and are potentially usable in a number of diseases. PMID- 10376414 TI - [Targeted and controlled release of drugs using magnetoliposomes]. AB - The present paper deals with a stable drug delivery system which allows to concentrate the drug in a desired site and its programmable release. For these purposes the present authors have suggested to use magnetoliposomes (liposomes with enwrapped magnetite particles in their bilayers) which are magnetosensitive and may be maneuvered to the given site in the organism. Due to the magnetite particles, which are strong microwave absorbers, magnetoliposomes can be heated to higher temperatures which may subsequently lead to a leakage of encapsulated drug. Influence of static magnetic field on liposomes in the blood-stream was analyzed theoretically. Fluorescence spectrophotometry was used to determine the relationship between doses of microwave radiation and the extent of drug release. PMID- 10376415 TI - [Inter-organ variability in enzymatic hydrolysis of ammonium salts in rats in vitro]. AB - The study evaluated the rate and kinetics of enzymatic hydrolysis of N-(2 benzoyloxyethyl)-alkyl-dimethylammonium bromides, potential easily biodegradable disinfectants of soft character. The products of enzymatic hydrolysis of the substrates under study, catalysed by microsomal esterase, included substituted substrates choline and benzoic acid which, as a hydrolytic product, was essayed by HPLC. The effect of the length of the alkyl chain of the individual homologues on the rate of enzymatic hydrolysis and their affinity to microsomal esterase of the rat liver and lung in vitro was examined. The structural modification (varying length of the aliphatic chain on the ammonium nitrogen of these compounds) was found to significantly influence the kinetics of enzymatic hydrolysis of the esteric bond. From the viewpoint of the rate of enzymatic hydrolysis no significant inter-organ variability was observed: in the liver as well as the lung the dependence of the rate of enzymatic hydrolysis on the length of the aliphatic chain possesses the shape of a falling hyperbole with the maxima for BCH2 > BCH4 > BCH8 = BCH10. The specific activity of both esterases ranges within 0.2-3.5 nmol.min-1.mg-1. From the viewpoint of affinity, a marked inter organ difference was manifested by 10 times higher affinity of the substrates to the lung microsomal esterase in comparison with the liver one. The effect of the length of the alkyl of the individual homologues on the affinity is of a non linear character also in this case. In both organs, a certain correlation was found between the rate of enzymatic hydrolysis and affinity to microsomal esterases. PMID- 10376416 TI - [Comparison of the effects of BI-6, a new asymmetric bipyridine oxime, with HI-6 oxime and obidoxime in combination with atropine on soman and fosdrine toxicity in mice]. AB - The therapeutic efficacy of the new asymmetric bispyridinium oxime BI-6 against acute toxicity of the highly toxic organophosphate soman and the organophosphorus insecticide fosdrin by means of affecting the LD50 values of these noxiores substances was compared with the effect of the hitherto most perspective oxime HI 6 and the classic obidoxime always in combination with the identical dose of atropine. At the equimolar level the effect of oxime BI-6 against fosdrin completely equals the effects of both oximes HI-6 and obidoxime. The effect of oxime BI-6 against soman is even more marked than the effect of HI-6 but this difference is not statistically significant. On the other hand, at the equi effective level, the effect of oxime BI-6 against soman is statistically significantly lower than the effect of HI-6, and against fosdrin it is even lower than the effect of both remaining oximes. The effects of the new oxime BI-6 equal, or slightly exceed the therapeutic effect of HI-6 but at the equimolar level only. At the equi-effective level which respects the toxicity of the oxime and is therefore more important for practical use, it is a therapeutically weaker reactivator of acetylcholinesterase than HI-6. PMID- 10376417 TI - [Generic drugs, alternatives, substitutes, interchanges]. AB - Linking up with the topical activities of the International Pharmaceutical Federation (FIP) in defining the concepts generic alternative, generic substitution, therapeutic alternative, therapeutic substitution and therapeutic replacement and the proposed policy, the paper describes the state and development in the field of generics. The attention is devoted to the position of generics in selected countries, their definitions, labelling, pricing and generic substitution. PMID- 10376418 TI - [The human hepatocyte: I. A model for studying metabolism and toxicity of xenobiotics]. AB - Isolated human hepatocytes have become one of the most attractive experimental approaches to the study of the specific metabolic functions of the human liver, interactions between liver cells and infectious agents, and metabolism and pharmacotoxicity of drugs. Particularly the primary cell culture model provides an in vitro system for investigating specific mechanisms in a precisely controlled conditions. In the present paper the legislative and ethical problems concerning availability of human liver samples, techniques developed for isolation, preservation and cultivation of hepatocytes are discussed. In addition, a comparison of human and rat hepatocyte models in the study of the metabolism and cytotoxicity of taxol is reviewed. PMID- 10376419 TI - Routine retroperitoneal drainage is not required for uncomplicated pelvic lymphadenectomy for uterine cancer. AB - Four hundred consecutive patients with endometrial carcinoma clinically confined to the corpus and cervix underwent extrafascial hysterectomy and pelvic lymphadenectomy by the author over an eight-year period of time. No patient had retroperitoneal drains, and only one patient (0.25%) developed a pelvic lymphocyst. Routine retroperitoneal drainage may be safely omitted in patients with uterine carcinoma undergoing hysterectomy and staging pelvic lymphadenctomy. PMID- 10376420 TI - The impact of pretreatment staging laparotomy on survival in locally advanced cervical carcinoma. AB - The purpose of this retrospective study was to determine the effect of pretreatment surgical staging on survival of patients with locally advanced cervical carcinoma. Two hundred and seventy-four women with cervical cancer stages IIB-IVA treated with primary radiotherapy comprised the study group. Eighty-nine patients underwent pretreatment staging laparotomy (group 1) and 172 patients underwent clinical staging (group 2). Thirteen patients underwent CT guided biopsy of paraaortic adenopathy. Paraaortic metastases were detected in 12.3% and intraabdominal metastases were found in 4.5% of patients in group 1. Extended field radiotherapy and/or systemic chemotherapy were given in these cases. The median survival of patients in group 1 was statistically longer than that of patients in group 2, 29 months vs 19 months, respectively (p=.01). Multivariate analysis controlling for both stage and age showed pretreatment staging laparotomy is a significant predictor of survival (p=.03). Our data suggest that surgical staging may be beneficial in patients with locally advanced cervical carcinoma. PMID- 10376421 TI - Recurrent borderline ovarian tumours after conservative management in women wishing to retain their fertility. AB - OBJECTIVE: To evaluate the rate of recurrence after conservative treatment of Stage 1 borderline ovarian tumors. METHODS: Retrospective case note review of all women with borderline ovarian tumours diagnosed at Aberdeen, U.K. and loannina, Greece between 1981-1993. Outcome measures comprised initial surgery, stage of disease, histological type, pregnancy rate, recurrence rate and disease-free interval. RESULTS: Conservative surgery for borderline tumours in young women permitted high conception rates. Recurrence of tumour was high in this group, 71%, and presented late. CONCLUSION: Prolonged intensive follow-up is required for women treated conservatively for borderline malignant ovarian tumours. PMID- 10376422 TI - Extramammary Paget's disease of the vulva: report of five cases and review of the literature. AB - Five patients with vulvar extramammary Paget's disease (EMPD) are reported. They accounted for 4.4% of all vulvar malignancies diagnosed in the south of Israel between 1961-1997. Mean age was 64.4 years and the predominant symptom was vulvar pruritus. Three patients had intra-epidermal lesions (managed by wide local excision or, at most, simple vulvectomy), one had a minimally invasive lesion (managed by simple vulvectomy) and one had EMPD with an underlying apocrine gland adenocarcinoma (managed by radical vulvectomy and bilateral groin dissection followed by pelvic radiotherapy). EMPD recurred in two patients: 1) local recurrence after simple hemivulvectomy for an intra-epidermal lesion was successfully treated by wide local excision; 2) widespread recurrence at distant skin sites after simple vulvectomy for a minimally invasive lesion was unsuccessfully treated with systemic chemotherapy. It is concluded that vulvar EMPD is an uncommon neoplasm that primarily affects postmenopausal women. Its histogenesis is uncertain and most commonly it is an intra-epidermal lesion. The high rate of recurrent disease remains a challenge for optimal management. PMID- 10376423 TI - Pelvic malignancy in female nonagenarians. AB - Reports about pelvic malignancy in female nonagenarians are scarce. Thirteen women 90 years of age and older were diagnosed with a malignant pelvic tumor between 1984 and 1996. The majority of these malignancies were gynecologic in origin. The development of a malignant pelvic tumor was, in our experience, associated with a poor prognosis. However, long-term cure after definitive treatment, as deemed feasible, is not precluded. PMID- 10376424 TI - Malignant trophoblastic disease following a twin pregnancy consisting of a complete hydatiform mole and a normal fetus and placenta. A case report. AB - We report an unusual pregnancy with a complete hydatiform mole coexisting with a normal fetus and placenta. This report stresses the importance of a correct diagnosis and the dilemmas the clinician is faced with when managing such a case. Malignant trophoblastic disease occurs in 55% of complete hydatiform mole and fetus. Two-thirds require combination chemotherapy. PMID- 10376425 TI - Diagnostic dilemmas and current therapy of Fallopian tube cancer. AB - Primary tubal cancer, unlike ovarian cancer, is not routinely suspected preoperatively, and thus diagnosis and therapy are delayed. We have recently encountered two cases in which primary Fallopian tube cancer masqueraded as other lesions. One presented as a pelvic inflammatory process, the second as cervical cancer. Primary Fallopian cancer should be suspected by the clinician, even if the presenting symptoms are atypical. Chemotherapy with taxol and cisplatin was instituted following debulking surgery. PMID- 10376426 TI - Intraepithelial neoplasia and early stage vulvar cancer. Epidemiological, clinical and virological observations. AB - OBJECTIVE: Evaluation of vulvar intraepithelial neoplasia (VIN) and early vulvar cancer risk factor occurrence, frequency, localization and development. STUDY DESIGN: A clinical study carried out on 293 women aged 23-76 years with VIN and vulvar cancer stage I and in a control group of 115 cytologically and colposcopically negative women. METHODS: Clinical, colposcopic and morphological evaluation of the localization of VIN and vulvar carcinoma stage I concomitant with intraepithelial neoplasia in other parts of the lower genital tract. Anamnestic inquiry regarding risk factors. In situ hybridisation technique for HPV detection. Thomson's method for blood serum vitamin A level assessment. RESULTS AND CONCLUSION: An increased frequency of VIN and Ca stage I, especially in young women, has been observed in the past 15 years. In a group of young women under 45 years of age, those lesions were multifocal in 43 cases (63.2%), and unifocal in 25 patients (36.8%). In women over 45 years of age, multifocal lesions occurred in 35 (31.8%), and unifocal in 75 patients (68.2%). HPV infections concomitant with VIN and vulvar cancer stage I occurred in 61.5% of young women and in 17.5% of older females. VIN and stage I vulvar carcinoma coexisted with cervical intraepithelial neoplasia and cervical and/or vaginal cancer in 14 women (7.9%). The risk factor for VIN and early vulvar carcinoma occurrence in young women was different than in older patients. Long-term follow up of VIN 1 and VIN 2 showed that in over 1/3 of cases the lesions were persistent or recurred after a transient remission. Progression depended not only on dysplasia stage, but also on histological pattern. PMID- 10376427 TI - Exaggerated placental site erroneously diagnosed as non-metastatic trophoblastic disease. A case report. AB - BACKGROUND: Exaggerated placental site (EPS) is classified as a non-neoplastic trophoblastic lesion, and histologically it consists of endometrial and myometrial invasion of intermediate trophoblasts and syncytiotrophoblasts and it differs morphologically from placental site trophoblastic tumors and placental nodules. The purpose of this report is to increase physicians' awareness of this lesion. CASE: A 48-year-old woman with post-molar rising betahCG titers and a clinical diagnosis of non-metastatic trophoblastic disease underwent hysterectomy. Final histopathology showed exaggerated placental site--a lesion often unfamiliar to clinicians. CONCLUSION: It is suggested that operative hysteroscopy may be useful in the diagnosis and management of EPS. PMID- 10376428 TI - Pilot study on the treatment of cyclical mastodynia with Quinagolide. AB - In a pilot clinical trial on 52 patients, 75 microg Quinagolide given once per day was administered for the treatment of cyclical mastodynia. Linear analogue charts were used for the assessment of response. Decrease in breast pain, heaviness, tenderness and serum prolactin level on the one hand, and increases in the serum estradiol and progesterone levels on the other hand were noted after 3 and 6 months administration, and were statistically significant. Statistical analysis was performed by Paired t-test or Wilcoxon test. One-Way Anova Repeated Measures and Wilcoxon test and analysis of Covariance model. The beneficial effect of Quinagolide also lasted after the cessation of treatment. Fourteen patients dropped out during treatment. Adverse effects like nausea, low blood pressure, dizziness and constipation were rarely reported. PMID- 10376429 TI - Sarcoma botryoides of the female genital tract: long-term results of conservative treatment followed by pregnancy. A case report. PMID- 10376430 TI - Ovarian tumors during childhood and adolescence. A clinicopathological study. AB - Ovarian tumors during the first two decades of life represent the most frequent tumors of the female genital tract at this age. Clinical and therapeutical particularities differentiate them from the same tumors of older women. During the period 1993-1997, 57 ovarian tumors were diagnosed and managed in the Divisions of Pediatric and Adolescent Gynecology of the 1st and 2nd Departments of Obstetrics and Gynecology, University of Athens. Benign tumors were found to be 77.2%, malignant 15.8% and borderline 7%. Germ cell tumors were the most common 49.1%, epithelial 35.1%, stromal 12.3%, lymphoma 1.75% and mixed tumors 1.75%. Of all the malignancies germ cell carcinomas consisted of 44.5%. PMID- 10376431 TI - Evaluation of the risk of cervical intraepithelial neoplasia and human papilloma virus infection in renal transplant patients receiving immunosuppressive therapy. AB - OBJECTIVE: To investigate the risk of cervical intraepithelial neoplasia and the coexistence of human papilloma virus (HPV) infection in renal transplant patients receiving immunosuppressive therapy. MATERIALS AND METHODS: Cervical Papanicolaou (Pap) smear and colposcopic examinations were performed in 48 renal transplant patients receiving immunosuppressive therapy. Microbiological and histopathologic findings were discussed. RESULTS: The patients were evaluated as to cervical neoplasia risk factors and the results were found to be statistically insignificant (p>0.05). Genital neoplasia was encountered in 20 of the 48 renal transplant patients. Koilocytosis developed in 6 out of 8 (75%) patients who were receiving high dose immunosuppressive therapy due to transplant rejection. HPV was found in 2 out of 48 patients; these 2 patients had koilocytosis in their cervical biopsies. The difference between the positive predictive value of colposcopic evaluation and the Pap smear was found to be insignificant (p>0.05). However, if colposcopy had not been performed in two cases of cervical intraepithelial neoplasia class I (CIN-I) and in one case of cervical microinvasive carcinoma, the cases would have been incorrectly diagnosed as normal by the false-negative results of the Pap smear. CONCLUSION: Renal transplant patients who were undergoing immunosuppressive therapy were found to be at increased risk of developing cervical intraepithelial neoplasia. All the patients using immunosuppressive agents should be followed-up by Pap smears every six months and by colposcopic evaluation every year. Avoiding high-risk sexual acts will decrease the risk of HPV transmission and the risk of genital neoplasia as well. PMID- 10376432 TI - Prognostic factors in stage IB-IIA cervical carcinomas treated with postoperative radiotherapy. AB - PURPOSE: This study investigated the prognostic significance of age, stage, tumor size, pelvic lymph node metastasis (PLM), surgical margin invasion, overall radiotherapy time (ORT), and interval between radiotherapy and surgery (IRS) in stage IB-IIA cervical carcinoma. METHOD AND MATERIALS: 100 patients treated with radical hysterectomy and postoperative radiotherapy were evaluated retrospectively. RESULTS: The 5-yr overall survival (OS), disease-free survival (DFS), and pelvic control rate (PC) were 83.6%, 82.8%, and 91.8%, respectively. PLM (p=0.008), IRS (p=0.01), ORT (p=0.007), and tumor size (p=0.028) were found to be significant on PC. PLM (p=0.04), ORT (p=0.04), and IRS (p=0.001) were significant on OS. PLM was significant (p=0.04) and IRS was marginally significant (p=0.06) on DFS. After multivariate analysis, PLM was significant on OS, DFS and PC. Recurrences were seen in 14 patients. CONCLUSION: According to this study PLM, IRS, and ORT are the most important prognostic factors. Recurrences outside the radiation volume leads to treatment failure. PMID- 10376433 TI - Lymphocyte-infiltrated FIGO Stage IIB squamous cell carcinoma of the cervix is a prominent factor for disease-free survival. AB - BACKGROUND: The data of histopathological factors affecting prognosis of patients with FIGO Stage IIB squamous cell carcinomas of the cervix are rarely known. The purpose of this study was to evaluate the histopathological factors rendering therapeutic failure in patients with FIGO Stage IIB. METHODS: Survival and prognostic factors were analyzed in 83 patients with FIGO stage IIB squamous cell carcinoma of the cervix treated with radical hysterectomy and retroperitoneal lymphadenectomy between 1980 and 1989. The relapse-free and overall survival rate was 61.4%. RESULTS: Univariate analysis demonstrated that the degree of lymphoreticular cells infiltrating the cervix, lymph node metastases, barrel shape of the cervix, lymph-vascular space invasion, more than three lymph nodes involved, and bulky tumor size (i.e. tumor size larger than 4 cm) were of prognostic significance in disease-free survival. However, pronounced lymphoreticular cells infiltrating the cervical cancer was the only independent factor to disease-free survival in patients with FIGO stage IIB squamous cell carcinoma treated with radical hysterectomy and retroperitoneal lymph node dissection using multivariate analysis. CONCLUSIONS: We concluded that the immune function of the host might play a crucial role in the process of anti-neoplasm even if local advanced carcinoma of the cervix is noted. Individual therapies may very well have limited efficacy based on the patient's tumor or the inherent host response. How to strengthen the power of the host's immune system may offer another thinking process to treat such a subgroup of patients. More studies are required. PMID- 10376434 TI - Expression of a local immune defense system in the female genital tract. An immunohistochemical study. AB - The aim of this study was to investigate the presence of a local immunological defense mechanism of the secretory IgA class in the female genital tract. MATERIAL AND METHODS: We studied by a streptavidin-biotin method the secretory component (SC) and IgA distribution in paraffin-embedded sections of 90 formalin fixed specimens. We studied 10 normal and 5 neoplastic cervical specimens, 20 normal, 10 hyperplastic endometrial specimens and 10 endometrial adenocarcinomas, 5 normal ovarian tubes and 30 ovarian epithelial neoplasms, serous and mucinous. A polyclonal SC and (Dako) and a mab IgA (Dako) was used and the reaction was scored from 1-3. RESULTS: Normal cervical mucosa and atrophic endometria were negative, while the basal portion of the endometrium, focally the proliferative glands, most of the secretory glands and most of the hyperplastic glands were positive for SC. IgA showed a similar distribution and a perivascular stromal reaction. Adenocarcinomas were positive for SC, but the intensity of the reaction was dependent on the differentiation of the tumors. Mucous and most serous neoplasms were negative for SC. IgA showed a similar reaction. CONCLUSION: There is evidence that the female genital tract has a local defensive immune system that may be hormone dependent. SC is a valuable marker of glandular differentiation. PMID- 10376435 TI - The role of Chlamydia trachomatis infection in cervical cancer development. AB - Previous Chlamydia trachomatis infection elevates expression of: 1. TGFalpha in CIN I, CIN II, CIN III; 2. HPV 16 in CIN I, CIN II, CIN III and invasive carcinoma; 3. Ki 67 in CIN III and invasive carcinoma. Chronic Chlamydial infection is very often associated with cervical hypertrophy. PMID- 10376436 TI - Radial scar of the breast. AB - Four cases of radial scar of the breast, primarily diagnosed as carcinoma are presented; the patients were found among 858 patients who were operated on at our Breast Unit over the last 4 years. The lesion was revealed on routine mammographic examination in 4 of our patients, while in the fourth it was found by palpation. In two of our patients mammographic examination revealed architectural distortion, in one patient micro-calcifications and in the fourth patient a stellate lesion was found. In the last two patients the lesion was localized before surgery with a hook wire. Diagnosis was established by histopathologic examination in all cases. The surgeon, the radiologist and the pathologist should be aware of this clinical entity which, in spite of its benign character, has the ability to simulate invasive carcinoma clinically, mammographically and histopathologically. PMID- 10376437 TI - Clinical assessment of EMA/CO induced DNA damage in peripheral blood lymphocytes of high-risk gestational trophoblastic tumor patients. AB - From 1989 to 1994, Etoposide, Methotrexate, Actinomycin-D, Cyclophosphamide, Vincristine, Folic acid (EMA/CO) regimen was administered to seven patients with high-risk gestational tumours according to the Bagshawe 1976 criteria. Peripheral blood lymphocytes were obtained from two of these seven high-risk gestational trophoblastic patients receiving the EMA/CO regimen, and damage levels of DNA during chemotherapy were assessed using SCGE (single cell gel electrophoresis) assay. Additionally, the efficacy, toxicity and clinical results of EMA/CO regimen were evaluated in patients with high-risk gestational trophoblastic tumours. Fever (71.4%), leukopenia (57%), increase in transaminase concentrations (57%), trombocytopenia (57%), and anemia (57%) were among the most frequent side effects of the EMA/CO regimen. All these toxic effects were reversible and there was no need to stop the therapy. EMA/CO is highly effective in patients with high risk gestational trophoblastic disease and its toxicity is predictable and reversible. Because of chemotherapy, DNA damage that is shown in peripheral blood lymphocytes, increases at the 8th day of the EMA/CO regimen. When DNA damage is higher in patients, the course of chemotherapy per each patient is shortened. When DNA damage is higher in the patients, the multisystem effects due to toxicity are more significant. The SCGE assay has many possibilities in such research and has proved to be a relatively simple, quick and sensitive technique. PMID- 10376438 TI - p53 staining as a prognostic indicator in endometrial carcinoma. AB - OBJECTIVE: To investigate p53 expression in endometrial cancer and its significance as a prognostic indicator. METHODS: Thirty-five consecutively surgically treated patients with endometrial cancer had their p53 expression studied by immunoperoxidase staining and quantified by lighted microscopic evaluation of the staining pattern. The determination of mean percentage of p53 expression was compared to prognostic indicators of endometrial cancer. RESULTS: p53 staining was detected in 20 of the 35 cases of endometrial carcinoma. Eleven of the 21 endometrioid tumors stained positive, while 9 out of 14 tumors with more aggressive histology stained positive for p53. If the grade I and II patients were taken into account as a whole, there was a statistically significant correlation (p<0.001) between the grade I and II patients and the grade III patients. The difference was statistically significant between stage I and III (p<0.05). The difference between lymphovascular space invasion and no lymphovascular invasion and p53 positivity was statistically significant (p<0.05). CONCLUSION: p53 expression is more common in more aggressive histologic subtypes than in endometrioid adenocarcinomas. Strong expression of p53 correlates with advanced stage and high grade and is detected more frequently in endometrial cancers with lymphovascular invasion. PMID- 10376439 TI - Nonpeptide vasopressin antagonists: a new group of hormone blockers entering the scene. AB - After the story of success of hormone blockers for catecholamines, aldosterone and angiotensin II and their successful implementation into clinical practice another endocrine cardiovascular system has come into focus. It has long been known, that the hormone vasopressin plays an important role in peripheral vasoconstriction, hypertension and in several disease conditions with dilutional hyponatremia in edematous disorders, like congestive heart failure, liver cirrhosis, SIADH and nephrotic syndrome. A series of orally active nonpeptide antagonists against the vasopressin receptor subtypes has recently been synthesized and is now under intensive examination. Nonpeptide V1a-receptor specific antagonists, OPC 21268 and SR 49059, nonpeptide V2-receptor specific antagonists, SR 121463 A and VPA 985, and combined V1a-/V2-receptor antagonists, OPC 31260 and YM 087, have become available for clinical research. AVP-V2 receptor antagonists lead to a dose-dependent diabetes insipidus in animals and man. The term aquaretic drugs (aquaretics) has been coined for these drugs to highlight their different mechanism compared to the saluretic diuretic furosemide. V1a-receptor antagonists might offer new therapeutic advantages in the treatment of vasoconstriction and hypertension. Combined V1a-/V2-receptor antagonists might be beneficial in the treatment of congestive heart failure. Early results are promising and now need to be confirmed in large clinical studies. PMID- 10376440 TI - T cell reactivity to DR*0401- and DQ*0302-binding peptides of the putative autoantigen IA-2 in type 1 diabetes. AB - Type 1 diabetes is thought to be an autoimmune disease mediated by T lymphocytes recognizing critical islet cell antigens. Recently, the tyrosine phosphatase like protein IA-2 was suggested as a putative autoantigen in type 1 diabetes since autoantibodies are detected in sera of diabetic patients and prediabetic subjects. Similarly, T cell responses of peripheral blood lymphocytes of type 1 diabetic patients to this protein have been described. Only very few data is available about immunodominant epitopes of IA-2 recognized by T cells. We have studied T cell responses in type 1 diabetic patients and age and partly HLA matched controls to IA-2 peptides designed to bind HLA risk alleles of IDDM as DR*0401 and DQ*0302. Both diabetic patients and controls responded to IA-2ic and some of the peptides. Three peptides of the C-terminal region of IA-2 were recognised by T cells of a fraction of diabetic patients but at least two of these peptides triggered also T cell responses in DR*0401/DQ*0302-matched controls. Most peptides bound to different HLA alleles ("promiscous binders"). The identification of autoantigenic epitopes may offer clues to related sequences e.g. of viral origin what relates to models of diabetes pathogenesis ("molecular mimicry"). Secondly, the design of antigen- or even epitope-specific immune intervention strategies aiming at tolerization of disease specific T cells in type 1 diabetes may profit from the knowledge of immunodominant T cell epitopes of a putative autoantigen. PMID- 10376441 TI - Helicobacter pylori associated gastric pathology in patients with type II diabetes mellitus and its relationship with gastric emptying: the Ankara study. AB - Helicobacter pylori (HP) is the most common cause of nonerosive nonspecific gastritis. Gastric and duadenal ulcer both are found to be associated with HP infection. Another consequence of HP infection is that it may progress to chronic atrophic gastritis which is a well recognized risk factor for adenocarcinoma of the stomach. So by extension, HP infection can be accepted as a risk factor for gastric cancer. From this aspect, identification of risk groups is increasingly important. It is well-known that patients with diabetes mellitus are more prone to infection. Besides this, presence of gastroparesis diabeticorum may lead to bacterial overgrowth in the upper gastrointestinal (GI) tract. The present crossectional study was planned to study the presence of HP infection in diabetic patients with alterations in upper GI motility and to compare the results with healthy control group. Group I consisted of 51 patients with type II diabetes mellitus (as defined by National Data Group criteria) without any dyspeptic symptoms. Twenty-five age-matched healthy people served as a control in group II. Radionuclide-labelled solid meals were used to calculate gastric emptying time (GET). According to the results, patients in group I were divided into two groups. Patients with prolonged GET were grouped as group IA, while group IB consisted of patients with normal or shortened GET. Presence of HP gastritis is determined by histopathologic examination of endoscopic biopsy specimen. The results showed that the prevalence of HP gastritis in group I and II were 80.4% and 56% respectively and the difference was significant statistically (p: 0.03). In group IA, the prevalence of HP infection was estimated to be 88.2%, while in group IB it was 76.5% but the difference was not significant (p: 0.31). We have not found any correlation between HbA1c levels and the presence of HP infection in both group IA and IB (p values 0.26 and 0.15 respectively). We conclude that the prevalence of HP gastritis is higher in asymptomatic diabetic patients compared with healthy people. But there is no association between the alterations in GET and the presence of HP gastritis as indicated by our results. So prolonged GET may not be regarded as a specific pathogenic mechanism or a cause of HP infection in NIDDM patients. PMID- 10376442 TI - Unique alterations of thyroid function parameters after i.v. administration of alkylating drugs (cyclophosphamide and ifosfamide). AB - Alkylating drugs (cyclophosphamide and ifosfamide) have been in clinical use for the treatment of malignant diseases in the past. They are most useful anticancer agents and cyclophosphamide is also widely used for its immunosuppressive properties. However the effect of alkylating drugs on thyroid hormone parameters have not been evaluated so far. Three groups of patients were prospectively evaluated: Group I: 15 patients with Wegener's granulomatosis and 4 patients with severe scleritis received a single dose cyclophosphamide (15 mg/kg bw/day) and 250 mg prednisone i.v. Group II: 9 patients with malignant lymphomas were treated according to the IMVP 16-protocol. Patients received daily ifosfamide 1000 mg/m2 from day 0 to 4 and vepesid from day 0 to 2. Patients did not receive corticosteroids additionally. Group III: 6 patients with a relapse of malignant lymphomas received ifosfamide 1.500 mg/m2/day from day 0 to 4 i.v. and dexamethasone 40 mg/m2 as well as ara-c and etoposid. All patients received mesna to prevent hemorrhagic cystitis and odansetran or metoclopramide as antiemetic drugs. Alkylating drugs were given as a one hour infusion. Thyroid hormone parameters were determined before and on day 1, 2, 3, 4 after drug administration. We observed a significant increase in T4 and fT4 concentrations and a concomitant fall in TSH in either group one day after the administration of alkylating drugs. The effect was most pronounced in group III: T4 increased from 113 +/- 8 nmol/L to 175 +/- 8 (normal: 58-154) and fT4 from 14.0 +/- 0.8 to 24.8 +/- 2.5 pmol/L (normal 10-25). TSH dropped from 1.27 +/- 0.16 to 0.33 +/- 0.07 mU/L (normal 0.3-4). All changes were significant: p < 0.001. Two of the six patients displayed biochemical hyperthyroidism. Also reverse T3 increased significantly. Two days after drug administration a gradual normalization occurred. However, T3, Tg, TBG, Transthyretin and albumin levels did not change throughout the study period. One patient with coexisting hypothyroidism, who received his last thyroxine substitution therapy one day before the administration of cyclophosphamide (as in group I), also demonstrated an increase in T4, fT4 and rT3 and a fall in TSH concentrations. I.v. administrations of cyclophosphamide and ifosfamide induce a transient increase in T4 and fT4 concentrations and a concomitant fall of TSH in the presence of normal Tg, T3 and thyroid binding protein concentrations. These data suggest, that the changes are not due to a release of thyroid hormones from the thyroid itself, but is likewise related to a release of thyroxine from cellular pools such as the liver. PMID- 10376443 TI - Correlates of long-term hypocortisolism after transsphenoidal microsurgery for Cushing's disease. AB - In Cushing's disease, selective total removal of a corticotroph tumor of the pituitary regularly results in subnormal ACTH- and cortisol plasma levels. The duration of secondary adrenocortical insufficiency varies widely, with an average of 17 months in our patients. The goal of this study is to elucidate the underlying causes for the variation in duration of postoperative hypocortisolism. In this retrospective study, we evaluated 35 patients with postoperative hypocortisolism lasting more than 36 months, and compared them to 51 patients with a duration of less than 36 months. Preoperative data, intraoperative findings, and postoperative results with follow-up evaluations are presented. Extensive pituitary exploration, medial localization of the tumor, and a higher age were associated with increased risk for isolated secondary long-term hypocortisolism. The histological examination of paraadenomateous tissue identified a significantly larger amount of Crooke's cells in long-term cortisol insufficient patients. Previous pituitary surgery increased the risk for hypopituitarism. In some of our patients, the long-term adrenocortical insufficiency resolved after a period of over five years. PMID- 10376444 TI - Evaluation of procollagen III peptide as a marker of tissue hyperthyroidism in long-term treated women with TSH suppressive doses of thyroxine. AB - Increased serum concentrations of the aminoterminal propeptide of collagen III (PIIINP) are found in overt hyperthyroidism. Thus, measurement of serum PIIINP might be useful as an early marker of tissue hyperthyroidism in patients with TSH suppressive thyroxine treatment. In a prospective study we evaluated female patients followed in the thyroid outpatient clinic. Serum PIIINP concentrations were analysed in three groups: patients with TSH suppressive thyroxine treatment for more than 6 months (n = 33, TSH < 0.1 mU/l), patients with thyroxine substitution for hypothyroidism for more than 6 months (n = 20, TSH 0.2-4.0 mU/l) and spontaneous hyperthyroid patients (n = 8, TSH < 0.03 mU/l, increased freeT4 and/or T3). Beside TSH, thyroid hormones and serum PIIINP we measured serum SHBG and a clinical score. Hyperthyroid patients had clearly elevated serum PIIINP and SHBG values and a higher clinical score when compared with other study groups (p < 0.001). Patients with TSH suppressive thyroxine treatment had higher fT4 and T3 concentrations than the thyroxine substitution group (fT4 22 +/- 4.8 pmol/l vs. 17 +/- 2.6 pmol/l, T3 2.2 +/- 0.4 nmol/l vs. 1.8 +/- 0.3 nmol/l, p < 0.001) and also elevated serum SHBG values (77.6 +/- 27.5 nmol/l vs. 58.4 +/- 18 nmol/l, p < 0.01). However, serum PIIINP concentrations and the clinical score were very similar in both thyroxine treated groups (PIIINP in TSH suppression group 3.0 +/- 0.67 microg/l vs. 2.8 +/- 0.65 microg/l in the substitution group, clinical score 2 +/- 1.8 pts. vs. 1.7 +/- 1.5 pts. p = n.s.). In conclusion, serum PIIINP is not a reliable early marker for detection of tissue hyperthyroidism in long-term thyroxine treated women with suppressed TSH. PMID- 10376445 TI - Finasteride treatment for one year in 35 hirsute patients. AB - This study was performed to confirm the favourable therapeutic effects of finasteride in hirsute women as described by previous publications of different research groups. Our study was a non-randomized, prospective clinical trial. Thirty five patients with hirsutism were included in the study. The patients received 5 mg finasteride orally once per day over a period of 12 months. Hirsutism score, FSH, LH, E2, total T, free T, androstenedione (A), DHEAS, 17 hydroxyprogesterone (17-OHP) and sex hormone-binding globulin (SHBG) levels were determined in all the patients before treatment and every 6 months during treatment. The modified Ferriman-Gallwey score decreased from a mean of 19.06 +/- 6.12 to 11.31 +/- 4.93 during the study. Clinical improvement in the degree of hirsutism was observed in 26 of 35 patients. The percent reductions in hirsutism scores (mean% +/- SD) at 6 and at 12 months were 25.8 +/- 11.3 and 41.3 +/- 18.5, respectively. During the finasteride therapy E2 and SHBG were increased significantly while DHEAS was decreased significantly at 12 months. There were no significant changes in the values of the other hormones and no serious side effects were observed in the study. In conclusion, finasteride is a well tolerated therapeutic agent without significant side pathological laboratory findings and can be used in the treatment of hirsutism. PMID- 10376447 TI - Flow cytometric cell cycle analysis of two subpopulations of porcine granulosa cells. AB - Two types of granulosa cells from 120 individual follicles (forty follicles in particular stages of development) were analysed by DNA flow cytometry to determine the percentage of cells with degraded DNA (apoptosis) and the cell cycle analysis. The distribution of cells in the cell cycle (G1, S, G2/M) was studied to show relation to location within a follicle and follicular development. Isolated granulosa cells were scraped from the vesicle walls with rounded-tip ophthalmological tweezers and separated into weakly-associated (AGc) and tightly-bound (MGc) according to Ford (1991) and Duda (1997). Granulosa cells after fixation in 70% ethanol and staining with propidium iodide (PI) were analysed. At least 20,000 events were collected in each specimen. The S-phase fraction (SPF) and apoptosis were calculated using the ModFit LT programme for the cell cycle analysis (Verity Software House Inc., USA). In AGc a population with degraded DNA was found, containing less fluorescence than the G1/G0 peak as shown in the DNA histograms. The percentage of apoptotic AGc ranged from 39.29 in small, to 58.9 in medium and was significantly higher than in large follicles (26.13%; p < 0.05). The percentage of apoptotic MGc was significantly lower than in the AGc (p < 0.05) and was equal to 3.78 in small, 0.10 in medium and 0.08% in large follicles. There are no significant statistical differences between the mean percentage of SPF in MGc of small and medium follicles (4.94, 7.25%). However, SPF was significantly lower in large follicles (1.31%). The number of SPF in AGc decreased during follicular development (35.92, 26.98, and 19.62%). Our data indicate lack of apoptotic cell death in MGc which seem to be more differentiated, and lose their mitotic potential. In AGc however, which are undifferentiated and undergo numerous mitosis, apoptosis was observed. PMID- 10376446 TI - A study of the renal sodium excretion during the normal menstrual cycle using method of passive leg rising. AB - 12 healthy women (age 18-38 years) were examined using the 2-hour's method of passive leg rising (PLR) in follicular (FP) and luteal (LP) phases of normal ovulatory cycle. Renal and hormonal response to PLR was investigated. There was a significant increase of diuresis (from 53 +/- 9 ml/h to 298 +/- 27 ml/h in FP, from 69 +/- 12 to 324 +/- 28 ml/h in LP) and natriuresis (from 4.5 +/- 0.9 to 9.8 +/- 1 mmol/h in FP, from 5.7 +/- 0.3 to 12.1 +/- 1.1 mmol/h in LP), simultaneously with a decrease of plasma renin activity (PRA) and plasma aldosterone (PA) in both FP and LP. Baseline PRA was mildly and PA was significantly higher in LP compared to FP. Urinary osmolarity, heart rate and systolic blood pressure dropped significantly. Renal and hormonal response to PLR were identical in the two phases of the menstrual cycle. Authors conclude that 1/PLR causes significant diuresis and natriuresis due to central volume expansion and may be used as a simple stimulating test of renal sodium excretion, 2/renal sodium retention does not occur in the LP of normal ovulatory cycle. PMID- 10376448 TI - Autoantibodies to adrenal cytochrome P450 antigens in isolated Addison's disease and autoimmune polyendocrine syndrome type II. AB - Adrenal P450 enzymes 21-hydroxylase (21OH), 17alpha-hydroxylase (17OH) and side chain cleavage enzyme (SCC) represent major target antigens in adrenal autoimmunity. To evaluate the diagnostic sensitivity of autoantibodies to recombinant adrenal antigens we established rapid and sensitive radioligand assays and compared the results with adrenocortical autoantibodies (ACA) as detected by the standard immunofluorescence test. A high prevalence of antibodies to 21OH (21OH-A) was observed in patients with isolated Addison's disease (IAD) and patients suffering from autoimmune polyendocrine syndrome type II (APS II). 21OH-A were found in 19 of 25 (76%) patients with IAD and in 34 of 40 (85%) patients with APS II. In contrast, antibodies to 17OH (17OH-A) as well as antibodies to SCC (SCC-A) were detected in 12 (30%) and 13 (33%) patients with APS II whereas only a few sera from patients with IAD had 17OH antibodies (n = 3) and SCC-A (n = 1), respectively (p < 0.0001). The majority of patients with 17OH A (83.3%) or SCC-A (76.9%) were also found positive for 21OH-A and all three antibody specificities were positively correlated with the presence of ACA. Among 52 sera with ACA 49 (94.2%), 11 (21.2%), and 9 (17.3%) were positive for 21OH-A, 17OH-A and SCC-A, respectively. By combination of 21OH-A with 17OH-A all ACA positive individuals were identified. The availability of recombinant steroid P450 enzymes made it possible to develop radiobinding assays which allow simple, sensitive and quantitative detection of autoantibodies to defined adrenal autoantigens. We here demonstrate that autoantibodies to 21-hydroxylase are sensitive markers for autoimmune Addison's disease with and without polyglandular failure. The presence of 17OH-A or SCC-A may suggest the coexistence of or progression towards polyglandular autoimmunity. PMID- 10376449 TI - Human pancreatic islet quality control: easy assessment of metabolic functions. AB - We describe simplified and rapid methods to assess islet function with the aim to develop better protocols for islet isolation and to determine islet characteristics before transplantation. These methods are also useful in the assessment of the potentially beneficial or deleterious effects of compounds added to the culture media in stimulation experiments. To this end, we took advantage of the multiscreen assay system produced by Millipore SA. This 96-well unit allowed the free-floating culture of islets on filter membranes, the rapid vacuuming and collection of conditioned media or reaction buffer and thus successive testing of the same number of islets, possibly at different culture times. We estimated islet viability by determination of the metabolic activity of cells, normal function of islets by their ability to metabolize glucose and to synthesize and secrete insulin and of nitrite release, a reflection of nitric oxide (NO) status of cells, which may be involved in a signaling pathway during glucose-stimulated insulin secretion or in cytokine inducible pathway. Assays may be performed either on selected islets or on aliquots of semi-purified preparations designated for grafting, allowing thus the rapid estimation of graft function of the entire preparation. This herein described system may be also extended to many other functional tests. PMID- 10376450 TI - Frequency of severe hypoglycaemia in type 1 and type 2 diabetes during conventional insulin therapy. AB - To determine the frequency of severe hypoglycaemia during conventional insulin therapy in juvenile-onset and adult-onset type 1 and in type 2 diabetes mellitus (DM), we retrospectively analysed the medical records of 165 Turkish diabetic patients who have been treated with conventional insulin. Patients were divided into 3 subgroups with respect to the type of diabetes: 33 had juvenile-onset Type 1 DM, 18 had adult-onset type 1 DM, and 114 had type 2 DM. The diabetic subgroups were found to be comparable with regard to mean frequency of severe hypoglycaemia (juvenile-onset type 1 DM: 0.20 episode x patient(-1) x year-1, adult-onset type 1 DM: 0.10 episode x patient(-1) x year(-1), type 2 DM: 0.15 episode x patient( 1) x year(-1)). Frequency of severe hypoglycaemia necessitating in-hospital treatment was 0.05 episode x patient(-1) x year(-1) for all diabetic subgroups. The data clearly indicate that the extent of the problem of severe hypoglycaemia during conventional insulin therapy in type 2 DM is comparable with both juvenile and adult-onset forms of type 1 DM in Turkish diabetic population. PMID- 10376451 TI - Cutaneous neuroimmunomodulation: the proopiomelanocortin system. A New York Academy of Sciences Conference, September 11-13, 1998, Munster, Germany. PMID- 10376452 TI - Occult dysplasia is disclosed by Lugol chromoendoscopy in alcoholics at high risk for squamous cell carcinoma of the esophagus. AB - BACKGROUND AND STUDY AIMS: Squamous cell carcinoma of the esophagus (SCCE) becomes symptomatic at a late stage when the disease is already advanced, and this contributes to its poor prognosis. Esophagoscopy of asymptomatic individuals exposed to known risk factors associated with the development of this cancer may facilitate the diagnosis of early cancerous or precancerous lesions; however, conventional esophagoscopy is not accurate enough. The aim of this study was to measure the value of Lugol chromoendoscopy of the esophagus (LCE) as an endoscopic technique to detect dysplasia in patients at risk. PATIENTS AND METHODS: We studied 190 male patients older than 35 attending an outpatient unit for alcoholics who consumed more than 80g of alcohol, more than 10 cigarettes and more than 500 ml 'mate' (a hot infusion of herbs) per day over 10 years. All underwent conventional upper gastrointestinal endoscopy followed by LCE, a spraying of Lugol 3% on the entire esophagus. All patients denied dysphagia. Biopsies were obtained from any unstained areas larger than 5mm and also from stained areas in all individuals. Biopsies were analyzed independently by two pathologists unaware of the biopsy sites. All conventional esophagoscopies showed normal mucosa, except for two suspicious small elevated lesions, confirmed histologically to be SCCE. These two cases were excluded from the statistical analysis. RESULTS: The LCE found unstained areas in 23 patients and a uniformly stained esophageal mucosa in the remaining 165. Biopsies taken from these 23 unstained areas showed dysplasia in six (two high grade and four low grade), and the ones from the 165 stained areas taken at the middle esophagus showed low grade dysplasia in seven. There was a high prevalence (6.9%) of dysplastic lesions in these individuals and occult dysplasia was significantly more frequent in unstained than stained areas (p = 0.0017). LCE showed a sensitivity of 46%, a specificity of 90%, a positive predictive value of 26% and a negative predicitve value of 96% when unstained areas were compared to stained ones. Agreement between two independent pathologists was high, with a kappa coefficient of 0.64. CONCLUSION: We concluded that individuals who abuse alcohol, smoke and consume 'mate' have a high prevalence of dysplastic lesions that can be better detected by LCE. Esophagi with unstained areas had an eight-fold higher chance of revealing dysplasia than the uniformly stained ones. LCE is an easy and inexpensive method. It improves the detection of dysplasia and should be added to conventional upper GI endoscopy in patients at risk for SCCE. PMID- 10376453 TI - Combined ligation and sclerotherapy versus ligation alone for eradication of bleeding esophageal varices: a randomized and prospective trial. AB - BACKGROUND AND STUDY AIMS: A number of trials have been reported in which a combination of ligation and sclerotherapy was compared with ligation alone, or with sclerotherapy alone. The present trial was carried out to assess whether the combined therapy might achieve more rapid eradication of bleeding esophageal varices. PATIENTS AND METHODS: One hundred and three patients with either active bleeding or stigma of recent bleeding from esophageal varices were randomly assigned to receive ligation plus sclerotherapy, or ligation alone. Ligation was performed with the technique introduced by Stiegmann. Sclerotherapy was carried out using low-volume (1 ml) 1% aethoxysclerol, which was injected into varices proximal to each ligature. Further treatment sessions were held seven days later, and then at two-week intervals, until eradication of the varices was achieved. Endoscopic follow-up examinations were carried out at three-month intervals, or immediately if there was any recurrent bleeding. The mean follow-up period was 14 months. RESULTS: There were no significant differences between the groups of patients compared with regard to the number of sessions required to eradicate the varices (2.4 +/- 0.7 in the combined group, and 2.3 +/- 0.7 in the ligation group; p>0.05). No significant differences were found between the groups with regard to recurrent bleeding (Fp = 2.882; p > 0.05). Three cases of recurrent bleeding (6%) from treatment-induced ulcers and two cases of recurrent bleeding (4%) from duodenal ulcers were observed with the combined therapy and ligation, respectively. No significant differences in the mortality were found between the groups (Fp = 1.145; p>0.05). Two percent of patients in the ligation group died due to bacterial peritonitis. CONCLUSION: Since ligation combined with low-volume sclerotherapy did not reduce the time required for variceal eradication, it can be concluded that the combined therapy is not superior to ligation alone. This mode of endoscopic therapy for the treatment of bleeding esophageal varices is therefore not recommended. PMID- 10376454 TI - Preoperative staging of esophageal carcinoma: miniprobe sonography versus conventional endoscopic ultrasound in a prospective histopathologically verified study. AB - BACKGROUND AND STUDY AIMS: Endosonographic staging of esophageal carcinoma may be limited by non-traversable tumor stenoses. Dilation of malignant esophageal strictures carries a significant risk of esophageal perforation. We therefore evaluated the use of ultrasonic miniprobes in the staging of stenotic esophageal carcinoma compared with conventional endoscopic ultrasound. PATIENTS AND METHODS: In a blinded, prospective study, which included histopathological evaluation, 53 consecutive patients (43 male, 10 female, mean age 61 years) with stenosing esophageal carcinomas were examined preoperatively. Endosonography was done using the optical GF-UM3 echo endoscope. If tumor strictures were not traversable with this instrument, a blind esophagoprobe, the MH-908 was used for endosonography. Miniprobe sonography (MPS) was done during esophagoscopy in all patients. The various imaging modalities were assessed in terms of complete tumor traversability and correct tumor staging. Every patient underwent surgical tumor resection. RESULTS: MPS of the esophagus and proximal parts of the stomach was possible in all 53 patients without prior dilation of tumor stenoses. Endosonography with the GF-UM3 instrument was precluded in 23 patients (43.4%) while in 20 of the latter patients the MH 908 esophagoprobe could be passed through tumor stenoses. The overall accuracy rates for depth of tumor infiltration (T) staging were: 62% (31/50) for endosonography (GF-UM3 plus esophagoprobe) and 86.8% (46/53) for MPS. The accuracy rates for T staging in tumors traversable both with the GF-UM3 echo endoscope and with miniprobes were 56.7% (17/30) for GF-UM3 and 80% (24/30) for MPS. The accuracy rates for T staging in tumors traversable only with the MH-908 esophagoprobe and with miniprobes were 70% (14/20) for the MH-908 and 95% (19/20) for MPS. With regard to the presence or absence of peri-esophageal metastatic lymph nodes (N staging), the accuracy rates were 83% (25/30) for MPS and 70% (21/30) for the GF-UM3, and 80% (16/20) for MPS and 70% (14/20) for the MH-908. CONCLUSION: Compared with conventional endosonography using 7.5-MHz large diameter instruments, MPS enables: a) safe passage through high-grade malignant esophageal strictures, achieving b) higher accuracy rates for T staging, and c) similar rates for N staging. The use of MPS can also represent an improvement in the comfort and safety of patients. Moreover, miniprobe sonography is highly cost-effective compared with conventional endosonography. Thus, MPS appears to be a valuable addition to the armamentarium for staging esophageal carcinoma. PMID- 10376455 TI - Use of the enteroscope for colo-ileoscopy: low yield in unexplained lower gastrointestinal bleeding. AB - BACKGROUND AND STUDY AIMS: The small intestine is a potential origin of bleeding in patients with unexplained gastrointestinal tract hemorrhage or iron-deficiency anemia. Most reports on the investigation of these patients describe the use of upper tract enteroscopy. The diagnostic yield of combined upper and lower enteroscopy has not been widely assessed and remains to be clarified. The aim of this study was to assess the benefit of lower gastrointestinal tract enteroscopy in occult digestive bleeding. PATIENTS AND METHODS: Between 1 December 1995 and 15 January 1998, 54 patients with gastrointestinal bleeding of unknown origin were prospectively studied using upper and lower video push enteroscopy (44 for chronic iron-deficiency anemia and 10 for unexplained gastrointestinal tract hemorrhage with no potential site having been identified by other investigations). Examinations were done using a Olympus video enteroscope (SIF 100) under general anesthesia in a one-day clinic. An upper tract examination was done first, directly followed by the lower tract investigation. RESULTS: The upper tract enteroscopy was successful in 53 patients (98%) and retrograde ileoscopy in 21 patients (39%). In 18 (38%) cases the technical failure resulted from the impossibility of intubating the ileocecal valve. A potential source of upper gastrointestinal bleeding was detected in 35% of patients with chronic iron deficiency anemia and in 20% of those with unexplained gastrointestinal tract hemorrhage. The most common lesion in the small bowel was angiodysplasia (25%). The lower tract video push enteroscopy disclosed 11 lesions in patients with chronic anemia. However the lesions, including two ileocecal valve cancers, were mainly located in the colon and had been missed by previous colonoscopy. No case of ileal lesion was detected in this group of patients. In patients with unexplained gastrointestinal tract hemorrhage, three lesions were detected but only one of these was in the ileum. Associated colonic and jejunal lesions were observed in three patients (5.5%). Overall, the diagnostic yield of lower video push enteroscopy was less than 2%. CONCLUSION: This prospective study has shown that using an enteroscope as a colonoscope in the management of patients with gastrointestinal bleeding of unknown origin is of little help. It might actually be more appropriate to perform a second colonoscopy. This however remains controversial and a prospective study is needed to answer that question. PMID- 10376456 TI - Management of foreign bodies in the gastrointestinal tract: an analysis of 104 cases in children. AB - BACKGROUND AND STUDY AIMS: Ingested foreign bodies may be managed by endoscopy, observation, or surgery. The aim of the study was to investigate the methods of removal of foreign bodies according to type and location, success rates, and complications. PATIENTS AND METHODS: The charts of 104 children who had ingested foreign bodies were retrospectively reviewed. RESULTS: Of the patients, 80 (76.9%) were managed endoscopically. The overall success rate for endoscopic management was 98.8%. There were no complications during endoscopic interventions. In 23 cases the foreign bodies spontaneously passed through the gastrointestinal tract (22.1%). Surgical removal of a foreign body was done in only one case (0.96%). The majority of the foreign bodies which were located in the upper gastrointestinal tract could be removed endoscopically regardless of the nature of the material. Foreign bodies in the small and large intestine tended to pass through spontaneously without complications. CONCLUSIONS: It appears that the endoscopic approach is the preferable method for the extraction of upper gastrointestinal foreign bodies in child patients because of its high success rate, and that foreign bodies in the small and large intestine tend to be passed spontaneously without complications. PMID- 10376457 TI - Value of brush cytology for dominant strictures in primary sclerosing cholangitis. AB - BACKGROUND AND STUDY AIMS: Around 10% of patients with primary sclerosing cholangitis (PSC) develop cholangiocarcinoma, which is cholangiographically often indistinguishable from a benign dominant stricture. The aim of the present study was to assess the value of brush cytology in discriminating between benign and malignant dominant strictures in primary sclerosing cholangitis. PATIENTS AND METHODS: The results of all brush cytology specimens from dominant strictures from patients with established primary sclerosing cholangitis, taken at endoscopic retrograde cholangiopancreatography between 1987 and 1996, were compared with the histological diagnosis or clinical status of the patients at least 2 years later. RESULTS: A total of 47 brush cytology samples, taken between 1987 and 1996, from 43 PSC patients could be included. Between 1993 and 1996, p53 immunocytochemical examination was done in 27 brush cytology specimens and K-ras mutation analysis in 25 patients. The sensitivity, specificity, positive predictive value, and negative predictive value of brush cytology for detection of malignancy were 60%, 89%, 59%, and 89%, respectively. These figures were not improved by adding the results of p53 and K-ras analysis. Logistic regression analysis did not reveal any additional benefit of p53 or K-ras analysis either. Prior stenting did not adversely affect specificity. CONCLUSIONS: The sensitivity and positive predictive value of brush cytology for dominant strictures in PSC are rather poor. The specificity and negative predictive value are reasonably good. There was no additional value from p53 immunocytochemistry and K-ras mutation analysis. Prior stenting did not affect the results. PMID- 10376458 TI - Virtual reality training in laparoscopic surgery: a preliminary assessment of minimally invasive surgical trainer virtual reality (MIST VR). AB - BACKGROUND: The "fulcrum effect" of the body wall on surgical instrument manipulation is a major hurdle for novice endoscopic surgeons. Virtual reality training has not previously been evaluated as a means to overcome this problem. MATERIALS AND METHODS: 16 participants with no experience of endoscopy were required to make multiple defined incisions under laparoscopic laboratory conditions within 2-minute periods. Half of the subjects were randomized to receive initial training on the Minimally Invasive Surgical Trainer, Virtual Reality (MIST VR) computer programme. RESULTS: Participants with MIST VR training made significantly more correct incisions (P = 0.0001) than the control group on test trial 1, and even after extended practice by both groups (P = 0.0001). They were also significantly more likely to actively use both hands to perform the endoscopic evaluation task (P = 0.01). CONCLUSIONS: Virtual reality training represents a potential, viable solution for junior endoscopists, for overcoming the "fulcrum effect", in a replicable, safe learning environment which allows objective and reliable quantification of skill levels by trainers. PMID- 10376459 TI - Water intubation of the sigmoid colon: water instillation speeds up left-sided colonoscopy. AB - BACKGROUND AND STUDY AIMS: Rapid passage through the sigmoid and descending colon is important during flexible colonoscopy, and colonoscopists have developed several techniques and tricks for achieving this. The present study was designed to explore the effect of instilling 200 ml of water into the first bend of the sigmoid on the speed of passage of the endoscope from the rectum to the left colonic (splenic) flexure. PATIENTS AND METHODS: A prospective study of 100 successive single-handed colonoscopies was carried out, using randomly either the water intubation technique (50 patients) or the traditional method (50 controls) to compare the time needed to pass the endoscope from the rectum to the left colonic flexure. RESULTS: The results indicate that water intubation allowed the endoscope to be advanced through the sigmoid and descending colon in a median time (fiftieth percentile) of 154.5 seconds, compared to 223.5 seconds using the traditional technique. Water intubation speeds up the insertion time by 31%. This difference was highly significant statistically (P<0.0001). The difference remained significant when the data for men and women were analyzed separately. There was no statistically significant difference in the formation of N loops, or in incidentally formed alpha loops between the two study groups. CONCLUSION: The water intubation technique is more efficient than the traditional method, particularly in difficult left-sided colonoscopies, but it is equally safe. PMID- 10376460 TI - Initial experience with laparoscopic ultrasound-guided radiofrequency thermal ablation of hepatic tumours. AB - BACKGROUND AND STUDY AIMS: Radiofrequency (RF) thermal ablation has been applied almost exclusively through the percutaneous approach under radiological/external ultrasound guidance. We have embarked on a programme of laparoscopic ultrasound guided RF ablation of hepatic tumours in view of the potential advantages of this approach, i. e. reduced heat sink effect, greater precision and improved assessment of the thermal ablative zone. PATIENTS AND METHODS: RF thermal ablation using the Zomed International generator and multielectrode probes in two patients with hepatoma arising on a background of cirrhosis and in eight patients with multiple deposits from primary colorectal cancer. RESULTS: Total ablation was performed in two patients with hepatoma and 7/8 patients with secondary deposits. Total ablation with a minimum of 0.5-cm margin was achieved in 32 lesions. No complications were encountered postoperatively and all patients were discharged within 2 days of the intervention. One patient in whom thermal ablation was not completed has since died of progressive disease, eight appear to be free of tumour (follow-up 6-20 months) but one patient has developed further secondary hepatic deposits. CONCLUSIONS: The initial experience with laparoscopic ultrasound-guided RF ablation of hepatic tumours indicates its safety and therapeutic potential in patients with inoperable hepatic tumours. PMID- 10376461 TI - Hereditary angioedema with gastrointestinal involvement: endoscopic appearance. AB - We present the first reported case of hereditary angioedema (HAE) with gastric involvement to be successfully evaluated by endoscopy both during and after an attack. A 31-year-old man who had a family history of angioedema was admitted to our hospital with complaints of abdominal pain and swelling of extremities. Computed tomography scan and endoscopy carried out during this attack revealed transient gastrointestinal wall edema which, along with decreased levels of serum C4 and C1 inhibitor, confirmed the diagnosis of HAE with gastrointestinal involvement. During the attack, the gastric mucosa was erythematous and edematous, and parts of its surface bulged into the gastric lumen, resembling a submucosal tumor, as a result of massive submucosal edema. During the healing process, a number of small nodules and raised erosions developed over the entire gastric mucosal surface after healing of prominent gastric edema. Within 55 days, the gastric mucosa had returned to normal. The endoscopic findings for the stomach in HAE have not, to our knowledge, been previously described. PMID- 10376462 TI - Screening for early esophageal cancer: whom and how? PMID- 10376463 TI - How far have we come with ultrasound miniprobes? PMID- 10376464 TI - Value of brush border cytology for dominant strictures in primary sclerosing cholangitis. PMID- 10376465 TI - Gastric ascariasis. PMID- 10376466 TI - Fast method of jejunal biopsy. PMID- 10376467 TI - Guide wire technique for difficult pharyngeal passage. PMID- 10376468 TI - Bowel gas explosion during argon plasma coagulation. PMID- 10376469 TI - A partially displaced percutaneous biliary metallic stent: endoscopic management. PMID- 10376470 TI - Minute invasive squamous cell carcinoma of the esophagus associated with head and neck cancer: a case report. PMID- 10376471 TI - Laparoscopic removal of an ingested foreign body that had migrated into the liver. PMID- 10376472 TI - A case of abscess formation in a remnant common bile duct after Roux-en-Y hepaticojejunostomy. PMID- 10376473 TI - Spontaneous disappearance of a gastric lipoma after endoscopic biopsy: report of an unusual case. PMID- 10376474 TI - Giant cavernous hemangioma of the esophagus: endoscopic and echo-endoscopic appearance. PMID- 10376475 TI - Artificial playing surfaces research: a review of medical, engineering and biomechanical aspects. AB - In this paper, current knowledge of artificial playing surfaces is reviewed. Research status in the fields of sports medicine, engineering and biomechanics is described. A multidisciplinary approach to the study of artificial sports surface properties is recommended. The development of modelling techniques to characterise fundamental material properties is described as the most appropriate method for the unique specification of material properties such as stiffness and damping characteristics. It is suggested that the systematic manipulation of fundamental surface material properties in biomechanics research will allow the identification of subject responses to clearly defined surface variation. It is suggested that subjects should be grouped according to characteristic behaviour on specific sports surfaces. It is speculated that future biomechanics research will identify subject criterion related to differing group responses. The literature evidence of interactions between sports shoes and sports surfaces leads to the suggestion that sports shoe and sports surface companies should work together in the development of ideal shoe - surface combinations for particular groups of subjects. PMID- 10376476 TI - Decreased chronotropic drive as an adaptation to chronic exercise; possible mechanisms. AB - Cardiovascular function was determined at rest and during exercise in twenty eight healthy, elite distance runners. Maximum heart rate was 184 +/- 6 b x min( 1), which was more than one SD lower than the age predicted value (p < 0.001), and an inverse correlation was observed between maximum heart rate and VO2max (r = - 0.82, p < 0.001). The most aerobically trained athlete, a 27 year old male, presented with a maximum heart rate of 139b x min(-1). Echo-cardiac ultrasound revealed unremarkable intra-cardiac dimensions and flow characteristics relative to other endurance-trained athletes. A decreased chronotropic drive may represent a favourable physiological adaptation to endurance exercise which increases cardiovascular efficiency. PMID- 10376477 TI - Alterations in cardiac morphology and function in elite multi-disciplinary athletes. AB - Early echocardiographic studies of left ventricular (LV) morphology and function focused on single discipline athletes, primarily endurance and strength trained. To date there are few studies examining multi-disciplinary trained athletes. The present echocardiographic study examined LV morphology and function in 18 elite triathletes (swimming, cycling, and running) and 11 elite modern pentathletes (running, swimming, shooting, fencing, and show-jumping) compared with age- and sex-matched controls. Elite triathletes demonstrated significantly (p < 0.05) increased LV wall thickness and cavity dimensions together with LV mass, both in absolute terms and scaled for body surface area, compared with controls. Elite modern pentathletes demonstrated significantly (p < 0.05) increased LV wall thickness with a non-significant increase in LV internal diameter. Despite significant LV enlargement, the distribution of hypertrophy and diastolic filling indices were normal in both triathletes and modern pentathletes and significantly increased in the triathletes. It is concluded that multi-disciplinary training results in variations in LV morphology. The inciting stimulus resulting in LV enlargement in triathletes is associated with prolonged endurance activity, together with an isometric component accompanying cycling. In contrast, elite modern pentathletes experience a reduced endurance component combined with a high isometric component associated with fencing. PMID- 10376478 TI - Pre-exercise ingestion of carbohydrate and transient hypoglycemia during exercise. AB - To study the occurrence and contributing factors of transient hypoglycemia after pre-exercise ingestion of glucose after a 4-hour fast, 19 well-trained cyclists ingested 50 grams of glucose dissolved in water around noon after having a normal breakfast. The ingestion of the glucose solution was followed by 30 minutes rest after which the subjects cycled for 40 minutes at 60% of the predetermined maximal power output. Every 10 minutes blood was sampled for determination of glucose, catecholamines, and insulin concentrations. In 6 subjects (hypo-group) plasma glucose levels dropped transiently below 3.0 mmol/l, while in the other 13 subjects (non-hypo group) plasma glucose level remained above this level. Although at the onset of exercise the plasma glucose levels were lower in the hypo-group, insulin levels were similar in both groups, suggesting a higher insulin sensitivity in the hypo-group. During exercise, norepinephrine was lower in the hypo-group, indicating a lower sympathetic activity in the hypo-group. The lowest plasma glucose levels in both groups were observed after 20 minutes of exercise, after which plasma glucose concentration returned to normal levels. It is concluded that pre-exercise carbohydrate ingestion after a 4-hour fast is sufficient to induce a transient hypoglycemia. The data suggest that the occurrence of hypoglycemia is determined by a combination of a high insulin sensitivity, a small amount of ingested glucose, and a low sympathetic activity. PMID- 10376479 TI - Effects of exercise load and breathing frequency on heart rate and blood pressure variability during dynamic exercise. AB - It is known that heart rate (HR) variability decreases with dynamic exercise, but there are only few studies on blood pressure (BP) variability with exercise loads and the effect of breathing pattern has never been investigated. Thus, we studied HR and systolic blood pressure (SBP) signals by spectral analysis (FFT), in 9 healthy subjects, at different breathing frequencies (0.15, 0.2, 0.3, 0.4, 0.5, 0.6 Hz), at rest and during 3 exercise loads (25, 50 and 75% VO2max). BP was measured with a non-invasive device (Finapres) and continuously recorded. The power spectrum of R-R period significantly decreased with exercise loads in the low frequency band (LF: 0.04-0.128 Hz) and in the high frequency band (HF: 0.128 0.65 Hz), but with breathing frequency only in the HF part of the spectrum. The power spectrum of SBP significantly increased with exercise loads in LF and HF bands, and decreased in HF band with increasing breathing frequency. R-R and SBP HF peaks were centered on breathing frequency peaks. Therefore, spectral analysis of HR and SBP confirm the withdrawal of vagal control during exercise, while mechanical effect of respiration on SBP persists. LF/HF ratio of R-R spectral components decreased with increasing load, whereas cardiovascular sympathetic activity is known to rise, suggesting that this ratio is not a good indicator of cardiovascular autonomic modulation during exercise. PMID- 10376480 TI - Physiological effects of variations in spontaneously chosen crank rate during sub maximal and supra-maximal upper body exercises. AB - The aim of the present study was to compare the physiological responses when the crank rate was chosen spontaneously (Ts) or set at +/- 10% (T-10%, T+10%) of the freely chosen rate, during two upper body exercises: i) a sub-maximal test (T(SUB)) in which intensities ranged from 50 to 80% (118.4 +/- 10.2 to 189.5 +/- 16.3 watts) of maximal power (MP) and ii) a supramaximal test (T(SUPRA)) in which power output was set at 110 and 120% (260.5 +/- 22.4 and 284.2 +/- 24.4 watts) of MP. Eight nationally and internationally ranked kayakers, aged 20 +/- 2 years, performed these tests in which power outputs were normalised in relation to the maximal power output determined during T(MP). In T(SUB+10%), oxygen uptake and ventilation were significantly (P< 0.05) higher than during T(SUBxS). In T(SUB+10%) and T(SUB-10%), energy expenditure was significantly (P<0.05) higher and gross and net efficiencies lower than during T(SUBxS). During T(SUPRA-10%) when the power output was set at 110% of MP, time to exhaustion was significantly higher (P<0.05) than during T(SUPRAxS). The findings of the present study suggest that upper body exercise performed on an ergocycle should be conducted using the freely and spontaneously chosen crank rate. PMID- 10376481 TI - Physiological characteristics of well-trained synchronized swimmers in relation to performance scores. AB - The purpose of this study was to examine the relationships between the physiological characteristics of synchronized swimmers and their performance scores. The subjects were 16 trained female synchronized swimmers with a mean age of 17.2 +/- 1.7 years (mean +/- SD). The examined variables were body dimensions (height, width, body mass, circumference of the body and segment length), body composition, isokinetic muscle strength of the elbow and knee during extension and flexion, abdominal muscle endurance, anaerobic power (leg extension power and peak blood lactate concentration), aerobic power (maximum oxygen uptake [VO2max], swimming velocity at the onset of blood lactate accumulation [OBLA-V]), and flexibility (standing trunk flexion, prone trunk extension and distance between the open legs). The performance scores had significant correlations (p < 0.05) with isokinetic muscle strength of the elbow extension and flexion, and the knee extension, abdominal muscle endurance, leg extension power, VO2max x wt(-1), OBLA V and distance between the open legs. However, no significant correlations were found between the performance scores and anthropometric variables. This study showed that the performance scores of synchronized swimmers correlated significantly with the functional aspects, and that muscle strength, muscle endurance and aerobic capacity seem to be particularly important determinants. PMID- 10376482 TI - Effects of ingesting a sports bar versus glucose polymer on substrate utilisation and ultra-endurance performance. AB - The purpose of this study was to determine whether the ingestion of a sports bar (BAR) containing a mixture of fat (7 g), protein (14 ) and carbohydrate (CHO; 19 ) improved ulta-endurance cycling performance compared to when an equicaloric amount of CHO was consumed. On two occasions separated by a minimum of 7 days, six highly trained (peak power output [PPO] 414 +/- 8 W) endurance cyclists rode for 330 min at approximately 50% of PPO (203 +/- 8 W) while ingesting either the BAR or just CHO, before performing a 400 k] time trial as fast as possible. Rates of fat oxidation were significantly greater at the end of the submaximal ride when subjects ingested the BAR compared to CHO (1.09 +/- 0.08 vs 0.73 +/- 0.08g x min(-1); P<0.05), and accordingly total fat oxidation was significantly higher (280 +/- 24 vs 203 +/- 25 g, P < 0.05). However, two subjects failed to complete the time trial after they consumed the BAR during the prolonged, submaximal ride, whereas all subjects managed to finish the time trial when ingesting CHO. In conclusion, ingestion of the sports bar enhanced fat metabolism during prolonged, submaximal exercise, but impaired subsequent high-intensity time-trial performance. PMID- 10376483 TI - The effects of B-sitosterol (BSS) and B-sitosterol glucoside (BSSG) mixture on selected immune parameters of marathon runners: inhibition of post marathon immune suppression and inflammation. AB - A pilot study was undertaken to investigate the effects of the intake of capsules containing the plant sterols and sterolins (BSS:BSSG mixture) on selected immune parameters of volunteers participating in an ultra-marathon in Cape Town, South Africa. Those runners having received active capsules (n=9) showed less neutrophilia, lymphopenia and leukocytosis when compared to their counterparts having received placebo capsules (n=8): the placebo treated individuals showed significant increases in their total white blood cell numbers as well as in their neutrophils (p=0.03 and 0.03 respectively). Furthermore, statistically significant increases within lymphocyte subsets were observed in the runners having received the active capsules: CD3+ cells increased (p=0.02) as did CD4+ cells (p=0.03). In parallel, the BSS:BSSG capsules decreased the plasma level of IL6 in the runners using the active capsules (p=0.08) and significantly decreased the cortisol: DHEAs ratio (p=0.03), suggesting that these volunteers had less of an inflammatory response and were less immune suppressed during the post-marathon recovery period. These findings justify further investigations into the use of the phytosterols to prevent the subtle immunosuppression associated with excessive physical stress. PMID- 10376484 TI - Conditioned patellar tendon-tap reflexes in patients with ACL reconstruction. AB - The patellar tendon-tap stretch reflexes were examined in six neurologically healthy young subjects (mean age = 27.1 yrs) who had developed persistent quadriceps strength deficit due to ACL reconstruction. Each subject was tested on two separate days. A specially designed apparatus was used to examine the unilateral and conditioned patellar tendon-tap reflex response utilizing three different conditioning intervals: 25 ms, 75 ms, 150 ms, and a unilateral reflex (control). Peak isometric force and contraction time were measured by using a strain gauge. Also, peak-to-peak EMG was measured by using bipolar surface electrodes which were placed over the middle of the rectus femoris. All data were collected with a microcomputer (sample rate = 1 kHz). Due to the small sample, the Kruskal-Wallis nonparametric analysis of variance was performed. All subjects demonstrated quadriceps strength deficits in the ACL leg when compared with the contralateral leg. This analysis determined that for both the ACL leg and the Non ACL leg the size of stretch reflex was facilitated at the long-latency conditioning intervals (75 and 150 ms), whereas it was inhibited at the short latency conditioning interval (25ms). However, the ANOVA model failed to reveal any differences in the conditioned stretch reflex between the ACL leg and the Non ACL leg. Also, no differences were observed at the unilateral condition. Taken together, these results indicate that ACL reconstruction results in significant strength deficits, but does not alter unilateral or conditioned reflex profiles. PMID- 10376485 TI - The one-hundredth anniversary of the first culture of a mollicute, the contagious bovine peripneumonia microbe, by Nocard and Roux, with the collaboration of Borrel, Salimbeni, and Dujardin-Baumetz. PMID- 10376486 TI - Campylobacter coli strains with enlarged flagellin genes isolated from river water. AB - A group of campylobacters isolated from river water were found to possess unusually large flagellin genes. Both phenotype and serology were consistent with identification as Campylobacter coli. Phylogenetic analysis of small (16S, rrs) and large subunit (23S, rrl) rRNA genes of a representative strain, NCTC 13006, demonstrated high levels of relatedness with C. jejuni and C. coli (99.1 and 98.3% similarity for 16S; 99.3 and 99.4% similarity for 23S). Large flagellin proteins were demonstrated by SDS-PAGE analysis. The flaA and flaB genes were sequenced and aligned with known campylobacter flagellin amino acid sequences. The encoded FlaA protein of the new group exhibited a high degree of divergence from other Campylobacter species. Within the central variable region of FlaA, a further hypervariable domain was identified containing characteristic repeated motifs. Separate pairwise alignments performed for the variable regions of the polypeptide indicated these large fla genes were more closely related to those of C. upsaliensis than to those of C. coli or C. jejuni. PMID- 10376487 TI - Bacteroides fragilis isolates compared by AP-PCR. AB - Bacteroides fragilis is a component of the normal intestinal flora and an important pathogen in nonintestinal endogenous infections. It has been associated with enteric infections and has already been detected in polluted water. In order to evaluate the genetic diversity of B. fragilis, a total of 31 isolates and two reference strains were examined. This collection included strains from nonintestinal infections [12], intestinal infections [5], intestinal microflora [10], aquatic environments [4], and the reference strains ATCC 25285 and ATCC 23745. DNA fingerprints were detected using two separate PCR reactions with different arbitrary primers. The computer-assisted system Taxotron (Institut Pasteur, Dr P. Grimont) was used to analyze the profiles obtained and dendrograms were generated. By using a distance of 0.65 as the threshold, two clusters (hereafter referred to as genotypes I and II) were defined. Strains of differents origins could be distributed into both genotypes. We were unable to detect any obvious correlation between a given genotype and the specific disease or the source of the corresponding strains. PMID- 10376488 TI - Epidemiological typing of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae isolates by pulsed-field gel electrophoresis and antibiotic susceptibility patterns. AB - Over a 16-month period, Klebsiella pneumoniae isolates from 102 patients admitted to a university hospital in Liege (Belgium) produced extended-spectrum beta lactamases. Pulsed-field gel electrophoresis of genome macrorestriction patterns with XbaI and antibiotic susceptibility patterns subdivided 39 isolates into eight clonally related groups. Two of them were implicated in the course of this outbreak. They were responsible for successive waves of infection or colonization in different wards of the hospital while the others were encountered sporadically. A beta-lactamase with an isoelectric point of 7.6 and consistent with type SHV-2 characterized all nine isolates chosen among both major groups. PMID- 10376489 TI - Microevolution through DNA exchange among strains of Neisseria meningitidis isolated during an outbreak in the Czech Republic. AB - Neisseria meningitidis is a highly variable bacterium. Indeed, N. meningitidis is naturally competent for transformation, and horizontal DNA exchange between strains may lead to mosaic genetic loci in N. meningitidis. We studied such an exchange in nature during an epidemic provoked by N. meningitidis. This epidemic started in the Czech Republic in 1993 and the original epidemic clone was shown to have the antigenic formula (serogroup:serotype:serosubtype) C:2a:P1.2,5. We analysed 145 meningococcal strains isolated in the Czech Republic between 1993 and 1997 using serological and genetic typing methods (multilocus enzyme electrophoresis and polymorphism of pilA and pilD genes). This analysis showed that genetic exchange between epidemic and endemic strains had occurred. Exchanges involved mostly surface-exposed structures such as the capsule, giving rise to new meningococcal variants. The expansion of these variants should be kept under close surveillance. PMID- 10376490 TI - CreA modulates the XlnR-induced expression on xylose of Aspergillus niger genes involved in xylan degradation. AB - The expression of the feruloyl esterase gene faeA, the alpha-glucuronidase gene aguA, the endoxylanase gene xlnB, and the beta-xylosidase gene xlnD from Aspergillus niger on xylose was studied in a wild-type strain and in a CreA mutant. A decrease in expression of all four genes was observed with increasing xylose concentrations in the wild-type strain, whereas expression levels in the CreA mutant were not influenced. The results in the wild type indicated that xylose concentrations higher than 1 mM resulted in repression of the expression of the xylanolytic genes tested mediated by the carbon catabolite repressor protein CreA. On xylose, the expression levels of the xylanolytic genes were therefore not only determined by induction via XlnR, but also by repression via CreA. The genes tested were not influenced to the same extent by XlnR or CreA, resulting in specific expression levels and patterns for each individual gene. PMID- 10376491 TI - HSa of Staphylococcus aureus, a new member of the HU family of bacterial histone like proteins. AB - Histone-like proteins of the HU family are small, sequence-independent DNA binding proteins that facilitate a variety of DNA transactions. Here we report isolation from cell-free extracts of Staphylococcus aureus of HSa, a new member of the HU family. The NH2-terminal amino acid sequence of HSa led to identification of the corresponding gene (hsa) using the genome sequence of S. aureus. HSa is 90 amino acids long (Mr = 9 620) and shares 64 to 80% identity with homologs found in the genus Bacillus. PMID- 10376492 TI - Use of vascular sealing devices (VasoSeal and Perclose) versus assisted manual compression (Femostop) in transcatheter coronary interventions requiring abciximab (ReoPro). AB - Transcatheter coronary interventions requiring abciximab (ReoPro) are associated with vascular access site complications. Several devices have been developed to aid in the closure of the femoral arteriotomy, including collagen plug devices (VasoSeal, AngioSeal), percutaneous suture closure (Perclose), and aids to manual compression (Femostop). In 185 patients who received abciximab plus aspirin and heparin for transcatheter coronary interventions, we compared femoral arteriotomy closure by three different methods: VasoSeal, Perclose, and Femostop. A composite endpoint of late complications defined as an access site-related bleed or hematoma that required blood transfusion or an extended hospital stay, pseudoaneurysm, arteriovenous fistula, arterial or venous thrombosis was compared. VasoSeal was initially successful in 41/52 patients (78.8%). The 11 patients who failed to have adequate hemostasis with VasoSeal required manual compression aided by Femostop, but had no late complications. There was one access site infection and one fatal retroperitoneal hematoma unrelated to the vascular access site (surgically explored). There were no late complications. Perclose was successful in 48/56 patients (85.7%). One Perclose failure required surgical repair for an extensive arteriotomy. The other Perclose failure required manual compression aided by Femostop, but had no late complications. There were no access site infections requiring intravenous antibiotics. There was one retroperitoneal bleed that extended the patient's hospital stay and for which a blood transfusion was required. Femostop was successful in 77/77 patients (100%). There were no infections. Late complications occurred in four patients. These included three episodes of bleeding or hematomas requiring blood transfusion, and one pseudoaneurysm. CONCLUSION: In patients receiving abciximab in addition to aspirin and heparin, VasoSeal and Perclose are at least as safe as Femostop when used to achieve homeostasis after sheath removal. VasoSeal and Perclose have a significantly lower initial rate of successful hemostasis than Femostop. The numbers of late complications between the VasoSeal, Perclose, and Femostop groups were not significantly different. In those patients in whom VasoSeal or Perclose failed, no late complications occurred. Access site infections were no different between VasoSeal, Perclose, and Femostop. PMID- 10376493 TI - Risk of distal embolization and infarction with transluminal extraction atherectomy in saphenous vein grafts and native coronary arteries. NACI Investigators. New Approaches to Coronary Interventions. AB - Lower success rates have been reported when treating high-risk lesions in saphenous vein grafts (SVGs) and native coronary arteries with balloon angioplasty. The transluminal extraction atherectomy catheter (TEC) has been proposed to reduce the incidence of distal embolization (DE) in subsets of high risk lesions. To define the utility of TEC in reducing the incidence of DE, all patients who were enrolled in the New Approaches to Coronary Interventions (NACI) Registry and had TEC planned as the sole treatment were studied (329 patients with 381 lesions). Of the lesions treated, 75.9% were in SVGs; 37.5% were thrombotic; and 15% were total occlusions. Adjunctive percutaneous transluminal coronary angioplasty (PTCA) was performed in 87.4% of lesions. Multivariate predictors of DE were: noncardiac disease, stand alone TEC, thrombus, and larger vessel size. DE was associated with an 18.5% in-hospital mortality vs. 3.0% without DE (P < 0.01) and a 25.9% MI rate vs. 5.0% without DE (P < 0.01). In conclusion, in this high-risk subset of patients, TEC is associated with an 8.3% incidence of DE with thrombotic and SVGs lesions. DE associated with TEC appears to carry high morbidity and mortality. Additional techniques to control DE are needed to reduce the frequency of complications in these patients. PMID- 10376494 TI - Managing the embolization problem during saphenous vein graft intervention. PMID- 10376495 TI - Percutaneous treatment of pseudoaneurysms and arteriovenous fistulas after invasive vascular procedures. AB - Development of femoral artery pseudoaneurysms and arteriovenous fistulas represents a continuing problem after vascular diagnostic and interventional procedures. For most patients, ultrasound-guided compression is an effective method of treating such complications. However, in patients requiring a continuous anticoagulant regimen, in those with large arteriovenous fistulas or in patients suffering from painful groin hematomas, compression repair is less successful. We therefore assessed the feasibility, efficacy, and long-term results of interventional percutaneous treatment of these complications. In a 40 month period, we treated 53 consecutive patients with 30 pseudoaneurysms, 21 arteriovenous fistulas, and 2 combined lesions. The intervention was successful in 47 patients: 32 lesions were treated by implantation of covered stents, 14 by embolization techniques, and 1 by a combined procedure, surgical repair being necessary only in 6 patients. After a clinical and ultrasonic follow-up of 301 +/ 280 days, we noticed four late stent occlusions, especially in patients with poor peripheral runoff. Lesions with a distinct connection channel to the vessel lumen should be treated by coil embolization. In lesions originating from the femoral bifurcation with a broad base, surgical repair is necessary. Stenting of the superficial femoral artery with poor runoff should be avoided. Our results suggests that percutaneous closure of false aneurysms and arteriovenous fistulas after invasive procedures with unsuccessful ultrasonic compression repair is an attractive alternative to surgical treatment. PMID- 10376496 TI - You broke it, you fix it: more cards up the sleeve of the catheter man. PMID- 10376497 TI - Clinical predictors of improved long-term blood pressure control after successful stenting of hypertensive patients with obstructive renal artery atherosclerosis. AB - Despite a high procedural success rate, long-term blood pressure control after successful renal artery stenting of hypertensive patients has been inconsistent. This most likely reflects the absence of clinical guidelines for the selection of patients likely to benefit from renal revascularization. A cohort of 150 consecutive hypertensive patients (mean age, 66.7 years; 86 women) with 180 renal artery lesions (> or =75%) underwent primary Palmaz stent deployment. Mean arterial blood pressure (MAP), serum creatinine, and antihypertensive medication requirements were monitored prospectively. Specific definitions of blood pressure cure, improvement, or treatment failure were followed. Renal artery duplex Doppler or angiography was performed to assess stent patency at a mean 13 months (range, 7-15 months). Multivariate logistic regression analysis was used to select clinical variables that best related to a beneficial blood pressure control at follow-up. The procedural success rate was 97.3% (146 patients) and major in-laboratory complications were infrequent (1.3%). Late MAP values in 127 patients (91%) fell from 110 +/- 13.7 to 97.6 +/- 10.6 mm Hg (P < 0.001); antihypertensive medication requirements decreased from 2.9 +/- 1.2 to 1.9 +/- 1.1 (P < 0.01). The 13-month stent restenosis rate defined by duplex Doppler or angiography was 12%. Multivariate logistic regression analysis identified a preprocedure MAP of >110 mm Hg (odds ratio, 2.9; P = 0.003) and bilateral renal stenoses (odds ratio, 4.6; P = 0.009) as predictors of a beneficial blood pressure response at follow-up. This study provides general preprocedure guidelines for the selection of hypertensive patients with atherosclerotic renal lesions likely to benefit from primary Palmaz stenting and confirms a high procedural success and low stent restenosis rate. PMID- 10376498 TI - Optimizing the benefits of renal artery stenting. PMID- 10376499 TI - A prospective randomized trial of early ambulation following 8 French diagnostic cardiac catheterization. AB - This prospective randomized study was done to assess the safety of 4-hr ambulation after diagnostic cardiac catheterization with 8 French sheaths and catheters. In this selected group of patients, we found that early ambulation could be done without an increase in access site complications. PMID- 10376500 TI - "So how do you want to do this one?". PMID- 10376501 TI - Acute occlusion of the left main coronary artery following intracoronary ultrasound examination. AB - Intracoronary ultrasound (ICUS) is generally considered as safe procedure, with a low complication rate. We describe a nearly fatal complication of a diagnostic ICUS study that was treated successfully with stent implantation in the left main coronary artery and discuss the indications and remaining risks of this procedure. PMID- 10376502 TI - Delayed development of a coronary artery pseudoaneurysm after angioplasty. AB - One month after a successful angioplasty, one of our patients developed a new aneurysm in the right coronary artery (RCA). The aneurysm was characterized as a pseudoaneurysm by the use of intravascular ultrasound (IVUS). A stenosis that was not well seen by angiography was better depicted by IVUS. Both the pseudoaneurysm and the stenosis were successfully treated with a second angioplasty and stenting with a covered stent. Delayed development of pseudoaneurysms after dissection is an uncommon, but possible complication after angioplasty. In this case IVUS was useful for accurate characterization of the aneurysm. The use of covered stents may become a clinically useful method for treating coronary pseudoaneurysms. PMID- 10376503 TI - Treatment of a coronary artery to superior vena cava fistula resulting from early closure of a Possis Perma-Flow graft. AB - The Perma-Flow graft used in bypass surgery achieves more complete revascularization when paucity of native conduits exists. We report a coronary artery to superior vena cava fistula as a complication of this graft, leading to severe right heart failure. The fistula was successfully occluded percutaneously, improving the patient's clinical situation. PMID- 10376504 TI - Myocardial ischemia resulting from spontaneous dissection in a patient with massive bilateral sinus of valsalva aneurysms. AB - We describe a patient with large sinus of Valsalva aneurysms involving both the left and right coronary sinuses. Spontaneous dissection of the left coronary artery occurred, causing unstable angina, a complication heretofore not associated with this disease. Successful surgical reconstruction of the aortic root, aortic valve replacement, and coronary bypass grafting were performed. Pathology revealed cystic medial necrosis. PMID- 10376505 TI - Left internal mammary artery graft perforation due to high-pressure stent deployment. AB - Perforation of newly placed left internal mammary artery (LIMA) grafts due to stent deployment is an infrequent but potentially dangerous complication of coronary interventions. It may lead to brisk hemorrhage and massive cardiac tamponade requiring emergent pericardiocentesis and surgery. We report a case of a LIMA graft perforation following stent deployment with a high-pressure balloon 12 days after surgery. The patient was treated with emergent pericardiocentesis, rapid autotransfusion of the pericardial aspirate into the systemic circulation, and surgical repair of the ruptured vessel. PMID- 10376506 TI - Primary angioplasty for acute myocardial infarction resulting from the simultaneous occlusion of two major coronary arteries. AB - Primary angioplasty for acute myocardial infarction is frequently performed. Not uncommonly, more than one occluded artery may be present. Usually only one is an acute event, the others being chronic occlusions. We encountered a patient who presented with two simultaneous occlusions; both were successfully recanalized. We discuss some observations that assisted us in devising our treatment strategy. PMID- 10376507 TI - Post-infarction ventricular septal defect: percutaneous transvenous temporary closure using a Swan-Ganz catheter. AB - We report on a case of temporary closure of a post-infarction ventricular septal defect (VSD) using a Swan-Ganz catheter through a femoral transvenous approach. This resulted in substantial improvement in the hemodynamic status of the patient. Six hr later, the patient underwent surgery for VSD closure. When immediate surgical intervention is not possible, it may be helpful to stabilize the patient until surgery can be performed. Thus, such a treatment has potential as a temporary measure for patients awaiting surgical repair of post-infarction VSD. PMID- 10376508 TI - Calcified splenic artery aneurysm discovered during coronary arteriography. AB - We present a case of calcified splenic artery aneurysm discovered during routine coronary angiography. Early awareness of these aneurysms is important since their rupture may cause serious complications. This case emphasizes the importance of attention to cardiac and extra-cardiac structures during coronary angiography. PMID- 10376509 TI - String-plucking as a mechanism of chordal rupture during balloon mitral valvuloplasty using inoue balloon catheter. AB - Percutaneous transvenous mitral commissurotomy using the Inoue technique was performed in a 59-year-old female with mitral stenosis and a severely calcified mitral leaflets. Although not entrapped in the subvalvular apparatus, the balloon catheter was deviated away from the mitral orifice-apex axis of the left ventricle during the inflation of the proximal balloon, which plucked and severed the chordae tendineae of the posterior mitral leaflet and resulted in severe mitral regurgitation. PMID- 10376510 TI - Intralobar pulmonary sequestration supplied by the right coronary artery. AB - Bronchopulmonary sequestrations are malformations that are often congenital; they consist of isolated nonfunctioning lung segments having no communication with functional tracheobronchial elements of the surrounding lung. They are supplied by single or multiple branches from the distal thoracic or proximal abdominal aorta, or from the celiac, splenic, intercostal, subclavian, or pulmonary artery. Due to the absence of ventilation, the lung tissue can become chronically infected. We describe an intralobar pulmonary sequestration with arterial supply from the right coronary artery. PMID- 10376511 TI - Hemodynamic rounds series II: hemodynamic effects of alcohol-induced septal infarction for hypertrophic obstructive cardiomyopathy. PMID- 10376512 TI - Radiation safety in the cardiac catheterization laboratory: basic principles. AB - The possibility of injury from X-rays is only one of the risks of working in a cardiac catheterization laboratory. This article introduces information that should help understand the physics and radiobiology associated with radiation protection. PMID- 10376513 TI - Local intramural heparin delivery during primary angioplasty for acute myocardial infarction: results of the Local PAMI Pilot Study. AB - The feasibility and safety of local heparin delivery during acute infarct angioplasty was evaluated in a prospective, multicenter, 120-patient series. Angioplasty was performed using standard techniques, after which heparin (4,000 U) was delivered locally; 25% of patients received stents. Procedural success was reported in 98% of patients; 6.7% of patients suffered death, reinfarction, recurrent ischemia, or stroke during the index hospitalization. The 6-month target vessel revascularization rate was 12.5%. Local heparin therapy with provisional stenting in acute myocardial infarction patients is safe, feasible, associated with a low rate of infarct artery revascularization at 6 months, and may potentially eliminate the need for systemic heparin following the procedure. PMID- 10376514 TI - An embolization containment device. AB - A coaxial catheter system for containment of distal embolization is described. Utilizing a novel 0.014" hypotube with a distal elastomeric occlusion balloon, the PercuSurge GuardWire functions as a guidewire while trapping distal embolization resulting from more proximal intervention. The particulate debris is evacuated with a single operator exchange aspiration catheter (Export catheter) prior to deflation of the distal occlusion balloon. This animal study confirmed the feasibility of concept. The system was easily delivered through tortuous coronary anatomy. The GuardWire served as an adequate rail for delivery of dilatation balloons and a multitude of stents. There was no evidence of deep wall damage from low-pressure inflation and apposition of the distal occlusion balloon. PMID- 10376515 TI - Effect of stent design on reduction of elastic recoil: a comparison via quantitative intravascular ultrasound. AB - The increase in minimum lumen diameter achieved by coronary stent placement can be further enhanced by reducing the immediate recoil that occurs after stent deployment. The effect of various stent designs-flexible coils, slotted tubes, and a locking stent-on minimization of postdilation stent recoil was evaluated using an in vitro model of circumferential compression. The stents were expanded to 7 atm (3.82 +/- 0.02 mm); as pressure was reduced, lumen diameter and cross sectional area (CSA) were determined by on-line intravascular ultrasound imaging (30 MHz) positioned inside the dilating balloon (n = 10-15 inflation-deflation cycles). Stent recoil was assessed by calculation of percent change in CSA from 7 atm to negative balloon pressure: -33.1 +/- 5.6% (GR-II) and -22.4 +/- 3.8% (Wiktor) in the coil stents; -20.0 +/- 4.2% (JJIS coronary), -8.4 +/- 2.6% (JJIS biliary), and -6.9 +/- 1.5% (Multilink) in the slotted tube stents; and -1.9 +/- 3.2% in the Navius ZR1 locking stent (P < 0.05 vs. Multilink, P < 0.0001 vs. others). A range of resistances to recoil is demonstrated by this model, with coil stent designs undergoing greater elastic recoil than slotted tube stent designs. The locking stent design demonstrated the greatest radial strength and the most reduction in elastic recoil. PMID- 10376516 TI - Stent deformation following simulated side-branch dilatation: a comparison of five stent designs. AB - We aimed to determine the effects of simulated stent side-branch dilatation and subsequent redilatation of the central lumen. Following coronary stent implantation it may be necessary to dilate through the side of a stent to maintain branch patency. Branch dilatation through the side of 3.5-mm-diameter stents (AVE GFX, beStent, Crown, MultiLink, and NIR) was simulated in a plexiglass phantom using 2.5-, 3.0-, 3.5-, and 4.0-mm balloons. In further experiments, the main lumen was redilated with a 3.5-mm balloon after 3.0-mm side branch dilatation. Thereafter, a 3.5-mm central and a 3.0-mm side-branch balloon were simultaneously inflated ("kissing balloons"). The larger the balloon size used for side-branch dilatation, the greater the distortion of the stent immediately distal to the side-branch, which for a 4.0-mm balloon ranged from 36% +/- 2% (Crown) to 65% +/- 6% (NIR). Central lumen redilatation or kissing balloons abolished this stenosis with little reduction of the side-lumen diameter. The main stent lumen compromise caused by side-branch dilatation can be abolished by main-lumen redilatation or by kissing balloons. PMID- 10376517 TI - Coronary artery-descending aorta connection: fistula or collateral. PMID- 10376518 TI - Anomalous coronary vessels. PMID- 10376519 TI - Active constriction of the persistent arterial duct explains its intermittent reappearance. PMID- 10376520 TI - Addendum to "Short-term angiographic and long-term clinical follow-up of a patient with a malexpanded vein graft stent". PMID- 10376521 TI - Serine protease inhibitor TPCK prevents Taxol-induced cell death and blocks c-Raf 1 and Bcl-2 phosphorylation in human breast carcinoma cells. AB - The mechanism of Taxol-induced apoptosis was investigated in MCF-7 human breast carcinoma cells. Taxol-induced apoptosis was associated with phosphorylation of both c-Raf-1 and Bcl-2 and activation of ERK and JNK MAP kinases. The serine protease inhibitor N-tosyl-L-phenylalanine chloromethyl ketone (TPCK) effectively blocked apoptosis, but N-p-tosyl-L-lysine chloromethyl ketone (TLCK), another serine protease inhibitor, was without effect. TPCK treatment also prevented phosphorylation of c-Raf-1 and Bcl-2 in response to Taxol treatment. The serine protease inhibitor did not alter JNK activity, but it enhanced Taxol-induced activation of ERK1/2. Treatment of cells with the inhibitor of MEK activation, PD98059, prevented Taxol-induced ERK activation both in the presence and absence of TPCK, but did not influence survival of either Taxol- or Taxol plus TPCK treated cells. In addition, PD98059 had no effect on c-Raf-1 or Bcl-2 phosphorylation. Thus, while the Taxol-induced phosphorylations of c-Raf-1 and Bcl-2 proteins appear to be coupled, these events can be disassociated from ERK1/2 activation. In summary, these findings suggest that phosphorylation of c Raf-1 and Bcl-2, but not ERK1/2, are important signaling events in Taxol-induced apoptosis of MCF-7 breast cancer cells and that a TPCK inhibitable protease(s) is required for these processes. PMID- 10376522 TI - NDF/heregulin-induced cell cycle changes and apoptosis in breast tumour cells: role of PI3 kinase and p38 MAP kinase pathways. AB - Neu differentiation factor (NDF)/heregulin activates ErbB2 via heterodimerization with the NDF receptors ErbB3 and ErbB4. Cells which express normal levels of these receptors are often growth stimulated by NDF, whereas SKBR3, and other ErbB2-overexpressing breast tumour cells are growth inhibited. We demonstrate here that in SKBR3 cells, NDF induces G1 progression but also causes a G2 delay from day 1 and apoptosis from days 2-3. G1 progression was associated with ErbB2 transactivation of ErbB3 and subsequent stimulation of the phosphatidylinositol 3 kinase (PI3K) pathway whereas apoptosis was dependent on p38 MAPK. Inhibition of ERK1/ERK2 had no effect on cell cycle progression or apoptosis. Activation of ErbB3 and PI3K was also seen with betacellulin (BTC) but not epidermal growth factor (EGF) and correlated with the growth effects of these ligands. All three ligands induced short-term activation of p38 MAPK in a c-Src-dependent manner. However, only NDF caused a second, c-Src-independent increase in p38 MAPK activity which was required for apoptosis. PMID- 10376523 TI - Transcriptional regulation by the carboxyl terminus of c-Myb depends upon both the Myb DNA-binding domain and the DNA recognition site. AB - The c-Myb protein binds to DNA, can regulate transcription, and is required for normal hematopoiesis in vertebrates. Either amino- or carboxy-terminal truncation of this protein is required for efficient oncogenic activation. Previous studies have shown that the carboxyl terminus of c-Myb that is deleted in v-Myb contains negative regulatory domains. We now demonstrate that specific mutations within this carboxyl terminus result in greater transcriptional activation than truncation of the entire carboxyl terminus. Furthermore, this increased transcriptional activation depends upon the presence of the highly conserved Myb DNA-binding domain and is also dependent upon the nature of the Myb-binding sites within the target promoter. In a similar fashion, an activating mutation within the heptad leucine repeat region of c-Myb that is also present in v-Myb functions only in conjunction with the Myb DNA-binding domain and with particular Myb binding sites. These results suggest a model in which multiple domains of the c Myb protein are highly interdependent for transcriptional regulation. These interactions are promoter-specific and are not well modeled by heterologous fusion proteins. PMID- 10376524 TI - Evidence of an interaction between Mos and Hsp70: a role of the Mos residue serine 3 in mediating Hsp70 association. AB - c-Mos is a germ cell-specific MAP kinase kinase kinase (MAPKKK) that plays an essential role during meiotic divisions of oocytes. c-Mos is a key component of an activity, cytostatic factor, required for metaphase II arrest of unfertilized eggs in vertebrates. To understand the regulation of c-Mos, we are investigating c-Mos-interacting proteins. We provide evidence that mouse c-Mos binds to Hsp70, a molecular chaperone. Hsp70 was found to associate with Mos ectopically expressed in COS-1 cells. Mos-Hsp70 complexes could be immunoprecipitated with both Mos and Hsp70 antibodies. Despite a low-abundance of Mos, the Hsp70 antibody immunoprecipitated Mos as the major protein. Of importance, the Mos protein present in anti-Hsp70 immunoprecipitates functioned as an active MAPKKK indicating that it is not grossly misfolded. It is known that c-Mos protein kinase activity in cell extracts of transfected COS-1 or NIH3T3 cells is labile. We found that the inclusion of adenosine triphosphate (ATP) in cell extracts protected against the loss of Mos kinase activity. In the absence of ATP from cell extracts, protein kinase activity of Mos was lost within 6 h on ice even though the Mos protein was not degraded and remained bound to Hsp70. Based on our identification of c-Mos-Hsp70 interaction, one of the roles of ATP may be to assist the regulation of c-Mos via ATP involvement in the protein-folding function of Hsp70 and possibly other molecular chaperones. We also detected by coimmunoprecipitation a physical association between endogenous c-Mos and Hsp70 in Xenopus eggs. To provide further evidence for the functional significance of Hsp70 interaction to Mos function, we show that the residue serine 3 in Mos, which is important for the regulation of protein kinase activity of Mos is also important for Hsp70 association. PMID- 10376525 TI - Death-associated protein kinase 2 is a new calcium/calmodulin-dependent protein kinase that signals apoptosis through its catalytic activity. AB - We have identified and characterized a new calcium/calmodulin (Ca2+/CaM) dependent protein kinase termed death-associated protein kinase 2 (DAPK2) that contains an N-terminal protein kinase domain followed by a conserved CaM-binding domain with significant homologies to those of DAP kinase, a protein kinase involved in apoptosis. DAPK2 mRNA is expressed abundantly in heart, lung and skeletal muscle. The mapping results indicated that DAPK2 is located in the central region of mouse chromosome 9. In vitro kinase assay revealed that DAPK2 is autophosphorylated and phosphorylates myosin light chain (MLC) as an exogenous substrate. DAPK2 binds directly to CaM and is activated in a Ca2+/CaM-dependent manner. A constitutively active DAPK2 mutant is generated by removal of the CaM binding domain (deltaCaM). Treatment of agonists that elevate intracellular Ca2+ concentration led to the activation of DAPK2 and transfection studies revealed that DAPK2 is localized in the cytoplasm. Overexpression of DAPK2, but not the kinase negative mutant, significantly induced the morphological changes characteristic of apoptosis. These results indicate that DAPK2 is an additional member of DAP kinase family involved in apoptotic signaling. PMID- 10376526 TI - Expression of dominant-negative ErbB2 in the mammary gland of transgenic mice reveals a role in lobuloalveolar development and lactation. AB - Overexpression of the receptor tyrosine kinase ErbB2/HER2/Neu (ErbB2) occurs in 15-40% of human breast cancers. To determine the function of ErbB2 signaling during normal mouse mammary gland development, we expressed a carboxyl-terminal truncated dominant negative allele of ErbB2 (ErbB2deltaIC) in the developing mouse mammary gland. Despite ErbB2deltaIC expression within mammary glands of pubescent virgin and pregnant mice, a phenotype was not observed until late in pregnancy. At 1 day post-partum, lactationally active, distended lobuloalveoli failed to form. This phenotype was exaggerated in multiparous females expressing ErbB2deltaIC. Immunohistochemical staining for ErbB2deltaIC revealed a concordance between high levels of ErbB2deltaIC protein expression and the absence of lactational products within the lumens of ErbB2deltaIC stained lobuloalveoli. These results demonstrate that ErbB2 signaling is required for proper mammary development and lactation at parturition. PMID- 10376527 TI - Role of the ATFa/JNK2 complex in Jun activation. AB - The ATFa proteins, which are members of the CREB/ATF family of transcription factors, display quite versatile properties. We have previously shown that they interact with the adenovirus E1a oncoprotein, mediating part of its transcriptional activity and heterodimerize with the Jun, Fos or related transcription factors, thereby modulating their DNA-binding specificity. In the present study, we report the sequence requirement of the N-terminal activation domain of ATFa and demonstrate the importance of specific threonine residues (Thr51 and Thr53) in addition to that of the metal-binding domain, in transcriptional activation processes. We also show that the N-terminal domain of ATFa which stably binds the Jun N-terminal kinase-2 (JNK2) (Bocco et al., 1996), is not a substrate for this kinase in vivo but, instead, serves as a JNK2-docking site for ATFa-associated partners like JunD, allowing them to be phosphorylated by the bound kinase. PMID- 10376528 TI - Prohibitin, a potential tumor suppressor, interacts with RB and regulates E2F function. AB - The retinoblastoma tumor suppressor protein and its family members, p107 and p130, are major regulators of the mammalian cell cycle. They exert their growth suppressive effects at least in part by binding the E2F family of transcription factors and inhibiting their transcriptional activity. Agents that disrupt the interaction between Rb family proteins and E2F promote cell proliferation. Here we describe the characterization of a novel interaction between Rb family proteins and a potential tumor suppressor protein, prohibitin. Prohibitin physically interacts with all three Rb family proteins in vitro and in vivo, and was very effective in repressing E2F-mediated transcription. Prohibitin could inhibit the activity of E2Fs 1, 2, 3, 4 and 5, but could not affect the activity of promoters lacking an E2F site. Surprisingly, prohibitin-mediated repression of E2F could not be reversed by adenovirus E1A protein. A prohibitin mutant that could not bind to Rb was impaired in its ability to repress E2F activity and inhibit cell proliferation. We believe that prohibitin is a novel regulator of E2F activity that responds to specific signaling cascades. PMID- 10376529 TI - Role of caspases and possible involvement of retinoblastoma protein during TGFbeta-mediated apoptosis of human B lymphocytes. AB - In this study, we investigated the involvement of caspases in TGFbeta-induced apoptosis in human B cells. Our results show that TGFbeta-mediated nuclear fragmentation, observed in the Epstein-Barr virus-negative Burkitt's Lymphoma cell line BL41, was abolished in the presence of the tripeptide caspase inhibitor zVAD-fmk or the specific caspase-3 inhibitor DEVD-fmk. Other apoptotic manifestations such as cell shrinkage, surface phosphatidylserine expression and chromatin condensation were strongly inhibited by zVAD-fmk but only partially by DEVD-fmk. This suggests that other caspases in addition to caspase-3 control these apoptotis-associated features. Specific activation of caspase-3 during TGFbeta-induced apoptosis was demonstrated by the DEVD-fmk-sensitive expression of the active p17 subunit of caspase-3 and by in vivo cleavage of PARP. In addition, TGFbeta treatment of BL41 promoted the expression of both dephosphorylated and truncated forms of the retinoblastoma protein. Inhibition of caspase-3 activity abolished both nuclear fragmentation and expression of the truncated retinoblastoma protein, without modifying the G1 cell cycle arrest induced by TGFbeta. Our data thus demonstrate that TGFbeta-induced apoptosis of lymphoma B lymphocytes is dependent on caspase activation and involves caspase dependent cleavage of the retinoblastoma protein. PMID- 10376530 TI - Bcl-2 targeted to the endoplasmic reticulum can inhibit apoptosis induced by Myc but not etoposide in Rat-1 fibroblasts. AB - Bcl-2 is a key inhibitor of a broad range of apoptotic pathways, yet neither the mechanism of action nor the role of Bcl-2 subcellular localization are well understood. The subcellular localization of Bcl-2 includes the mitochondrial membrane as well as the contiguous membrane of the endoplasmic reticulum and nuclear envelope. Most studies suggest that the ability of Bcl-2 to confer cell survival is dependent upon its localization to the mitochondria. In this manuscript, we show that Bcl-2 targeted to the endoplasmic reticulum can inhibit Myc-, but not etoposide-induced apoptosis in the Rat-1 fibroblast cell line. By contrast, wild type Bcl-2 can inhibit apoptosis triggered by either death agonist. We further show both Myc and etoposide trigger disruption of mitochondrial membrane potential (MMP) and induce poly-ADP ribose polymerase (PARP) cleavage, but release of calcium was not evident. Bcl-2 abrogates apoptosis at or upstream of MMP depletion showing that Bcl-2 does not have to reside at the mitochondria to prevent apoptosis. These results further elucidate the biochemical events associated with Myc- and etoposide-induced apoptosis and significantly advance our understanding of Bcl-2 function. PMID- 10376531 TI - Expression and role of PML gene in normal adult hematopoiesis: functional interaction between PML and Rb proteins in erythropoiesis. AB - The expression of the PML gene was investigated in purified early hematopoietic progenitor cells (HPCs) induced to unilineage erythroid or granulocytic differentiation. PML mRNA and protein, while barely detectable in quiescent HPCs, are consistently induced by growth factor stimulation through the erythroid or granulocytic lineage. Thereafter, PML is downmodulated in late granulocytic maturation, whereas it is sustainably expressed through the erythroid pathway. In functional studies, PML expression was inhibited by addition of antisense oligomers targeting PML mRNA (alpha-PML). Interestingly, early treatment (day 0 HPCs) with alpha-PML reduced the number of both erythroid and granulocytic colonies, whereas late treatment (day 5 culture) reduced erythroid, but not granulocytic, clonogenesis. These findings suggest that PML is required for early hematopoiesis and erythroid, but not granulocytic maturation. The pattern of PML expression in normal hematopoiesis mimics that of retinoblastoma pRb 105. Combined treatment of HPCs with alpha-PML and alpha-Rb oligomers inhibited both PML and Rb protein expression and completely blocked erythroid colony development. Furthermore, PML and pRb 105 were co-immunoprecipitated in cellular lysates derived from erythroid precursors indicating that this functional interaction may have a biochemical basis. These results suggest a key functional role of PML in early hematopoiesis and late erythropoiesis: the latter phenomenon may be related to the molecular and functional interaction of PML with pRb 105. PMID- 10376532 TI - Cyclin D1 amplification is independent of p16 inactivation in head and neck squamous cell carcinoma. AB - Progression through the G1 phase of the cell cycle is mediated by phosphorylation of the retinoblastoma protein (pRb) resulting in the release of essential transcription factors such as E2F-1. The phosphorylation of pRb is regulated positively by cyclin D1/CDK4 and negatively by CDK inhibitors, such as p16 (CDKN2/MTS-1/INK4A). The p16/cyclin D1/Rb pathway plays a critical role in tumorigenesis and many tumor types display a high frequency of inactivation of at least one component of this pathway. In order to determine the overall contribution of these three components to progression of head and neck squamous cell carcinoma (HNSCC), we examined p16 inactivation, cyclin D1 amplification, and pRb expression in 23 primary HNSCC tumors and five cell lines. p16 inactivation was detected in 19/23 (83%) primary tumors by detailed genetic analysis and was confirmed by immunohistochemistry (IHC). Absence of Rb protein expression indicative of pRb inactivation was identified in 2/23 (9%) tumors. In this set of tumors, there was a perfect inverse correlation between p16 and pRb inactivation. Using fluorescence in situ hybridization (FISH) cyclin D1 amplification was identified in 4/5 (80%) cell lines and 4/11 (36%) primary tumors. However, 2/4 cell lines and all four primary tumors with cyclin D1 amplification contained a concomitant alteration of p16. Therefore 21/ 23 (91%) of primary HNSCC contained at least one alteration in the p16/cyclin D1/Rb pathway. Although p16 and Rb alteration are apparently exclusive, cyclin D1 amplification occurs concomitantly with the loss of p16 suggesting an additional role for this amplification in HNSCC. PMID- 10376534 TI - Changes for Health Physics On-Line. PMID- 10376533 TI - Identification and temporal expression pattern of genes modulated during irreversible growth arrest and terminal differentiation in human melanoma cells. AB - Abnormalities in differentiation are common occurrences in human cancers. Treatment of human melanoma cells with the combination of recombinant human fibroblast interferon (IFN-beta) and the antileukemic compound mezerein (MEZ) results in a loss of tumorigenic potential that correlates with an irreversible suppression in proliferative ability and induction of terminal differentiation. It is hypothesized that this is associated with the differential expression of genes that may directly regulate cancer cell growth and differentiation. To define the relevant gene expression changes that correlate with and potentially control these important cellular processes a differentiation induction subtraction hybridization (DISH) scheme is being used. A temporally spaced subtracted differentiation inducer treated (TSS) cDNA library was constructed and differentially expressed DISH clones were isolated and evaluated using a high throughput microchip cDNA (Synteni) array screening approach. Verification of differential gene expression for specific cDNAs was confirmed by Northern blotting. The temporal kinetics of regulation and the expression pattern of DISH genes were also evaluated by microchip cDNA array screening. Using this approach with 1000 DISH cDNA clones (approximately 10% of the DISH library) has resulted in the identification and cloning of both 26 known and 11 novel cDNAs of potential relevance to growth control and terminal differentiation in human melanoma cells. PMID- 10376535 TI - In-situ Object Counting System. PMID- 10376536 TI - Radiation protection information: can you trust the government's risks or risk the government's trust?: 1997 G. William Morgan lecture. AB - Public acceptance of information concerning radiation risks has been impacted by the erosion of trust in government agencies and by societal images that personify radiation or its effects in terms of monsters and ogres. The loss of trust in government agencies, particularly the Atomic Energy Commission and later the Department of Energy, has been influenced by a number of key events and individuals. Examples of these are given, including the anti-Viet Nam war movement, the Watergate incident, the activities of the Union of Concerned Scientists, Ralph Nader and the Critical Mass movement, the claims of Ernest Sternglass, and the widely publicized views of John Gofman and Arthur Tamplin. The use of negative images, pictures, and symbols in the mass media has reinforced the public perception of radiation as a thing to be feared. There is growing evidence that the public perception of radiation risks is related more to mistrust and negative images than it is to the technical information health physicists provide or to the issue of whether or not the linear no-threshold theory of radiation risks is correct. Attempts by federal agencies to regain public trust in radiation risk information generated by health physicists or other radiation scientists appear to be largely unsuccessful. If health physicists hope to be successful in changing such public perceptions, they may have to focus efforts on the next generation and concentrate on assuring that elementary and secondary school children receive sound instruction on radiation risks. Additional research at the molecular biology level is needed to elucidate the risks, if any, at low doses so that the practice of extrapolating low dose responses from high dose data can be eliminated. PMID- 10376537 TI - Induction of apoptosis by beta radiation from tritium compounds in mouse embryonic brain cells. AB - Induction of apoptosis by tritium exposure was investigated in both cultured embryonic mid brain cells and brain sections of embryos and of newborns in mice. In the cultures of mid brain cells, addition of methyl-3H-thymidine (3H-TdR) (21 kBq mL(-1)) and tritiated water (5.616 MBq mL(-1)) induced late appearances and low percentages of apoptosis when compared to x-irradiation at the ID50 dose, the inhibitory dose that reduced cellular differentiation by 50% of the control. A significant increase in p53 protein was detected about 2 h before the marked appearance of apoptosis. The pregnant mice were given an intraperitoneal injection of tritiated water at the concentration of 481.8 kBq g(-1) of body weight on gestation day 12.5, by which treatment behavioral changes in the offspring occurred. Increased apoptotic cells were observed in the neural tube of embryos from 1 d after the injection to 1 wk postnatal age. Apoptosis induced by x-rays appeared 2 h after irradiation, with a peak at 4 h. Increase of apoptotic cells was also found in the brain cortexes of newborns. The percentage of apoptosis in the brain was higher in the prenatal tritiated water exposed mice than in the prenatal x-irradiated mice. Possible mechanisms on apoptosis and its relation to the higher relative biological effectiveness value of tritium beta rays are discussed. PMID- 10376538 TI - The effect of lung deposition patterns on the activity estimate obtained from a large area germanium detector lung counter. AB - Lung counters for in vivo detection of low energy photon emitters are typically calibrated using phantoms containing lung tissue equivalent material with the radioactivity homogeneously distributed throughout the material. If the activity in a measurement subject is heterogeneously distributed, the activity estimate for that subject will be uncertain due to the assumptions of distribution. The magnitude of the uncertainty for a four-detector germanium array, using the Lawrence Livermore National Laboratory torso phantom with a newly designed lung set that allows the activity to be localized in one or more of 16 areas, was estimated. The results show that detector arrays will reduce the uncertainties arising from the geometry of the lung deposition compared to single detectors. The estimated activity of an internal deposition that emits 17.5 keV photons can be overestimated by a factor of three, or underestimated by a factor of infinity (i.e., the activity is missed completely). As the photon energy rises to 59.5 keV the uncertainty in the activity decreases so that the maximum overestimate (underestimate) will be a factor of two (five). As the energy rises to 344.3 keV only the maximum underestimate changes: it becomes a factor of three. PMID- 10376539 TI - Medical radiation exposures for diagnostic radiology in Malaysia. AB - The medical radiation usage for diagnostic radiology in Malaysia (a Level II country) for 1990-1994 is reported, enabling a comparison to be made for the first time with the United Nations Scientific Committee on the Effects of Atomic Radiation Report. In 1994, the number of physicians, radiologists, x-ray units, and x-ray examinations per 1,000 population was 0.45, 0.005, 0.065, and 183, respectively. (Level I countries had averages of 2.6, 0.072, 0.35, and 860, respectively). In 1994, a total of 3.6 million x-ray examinations were performed; the annual effective dose per capita to the population was 0.05 mSv, and the collective effective dose was 1,000 person-Sv. Chest examinations contributed 63% of the total. Almost all examinations experienced increasing frequency from 1990 to 1994 except for barium studies, cholecystography, and intravenous urography ( 23%, -36%, -51%). These decreases are related to the increasing use of ultrasound and greater availability of fiberoptic endoscopy. Notable increases during the same period were observed in computed tomography (161%), cardiac procedures (190%), and mammography (240%). In order to progress from Level II to Level I status Malaysia needs to expand and upgrade radiological service in tandem with the health care development of the country. PMID- 10376540 TI - An environmental radionuclide baseline study near three Canadian naval ports. AB - This paper summarizes an environmental radionuclide baseline study undertaken for the Department of National Defence in Canada. The purpose of the project was to establish levels of radionuclides present in the environment around areas where nuclear propelled vessels may be berthed. Specifically, this report describes environmental baselines near Halifax (Nova Scotia), Esquimalt (British Columbia), and Nanoose Bay (British Columbia). Valued ecosystem component samples were taken from dairy farms, beef producers, market gardens, vegetables, tree fruits and berries within the study areas, as well as marine bivalves (mussels and clams), salmon, seaweed, and food from native fisheries. Numerous naturally occurring isotopes were detected and quantified. The only non-naturally occurring isotope positively identified was in the form of trace quantities of 131I, measured in the Halifax study zone (attributed to local hospital cancer therapy). 137Cs is the only other anthropogenic radionuclide detected. Its origin may be the combination of fallout from the Chernobyl accident and fallout from atmospheric nuclear weapon tests. The results indicate that nuclear-powered vessels have not resulted in activity levels that would contribute a significant radiation exposure to the public, the biota, and the environment within the three study zones. PMID- 10376541 TI - Survey of 222Rn concentrations in the air of a tunnel located in Nagano City using the solid-state nuclear track detector method. AB - The survey of 222Rn concentration in the air of tunnels constructed during World War II has been performed using a solid-state nuclear track detector technique. For the practical application of this technique to the determination of 222Rn concentrations in air, some basic properties were experimentally examined on the cellulose nitrate film, Kodak LR 115 type II. The calibration coefficient of the cellulose nitrate film used is determined from a correlation between the 222Rn concentration in air and the observed number of perforated etched tracks for widespread radon concentrations. The slope of the linear relationship observed yields a calibration coefficient of (0.00209 +/- 0.00018) tracks cm(-2) (Bq m(-3) h)(-1). From the survey of 222Rn concentration in the air of tunnels, the concentration of several thousand Bq m(-3) was observed at the inner most area of the tunnel, and the seasonal variation was clearly observed. The exponential distribution of radon concentration as a function of distance from the openings of the tunnel suggests that the radon concentration in the tunnel is basically governed by diffusion and mixing of radon gas with air. PMID- 10376542 TI - Soil concentration, vertical distribution and inventory of plutonium, 241Am, 90Sr and 137Cs in the Marche Region of Central Italy. AB - Soil concentrations of 239+240Pu, 238Pu, 241Am, 90Sr, and 137Cs are investigated in the Marche Region of Central Italy. Mean values in uncultivated soils (0.721 +/- 0.456, 0.023 +/- 0.014, 0.241 +/- 0.165, 5.40 +/- 3.32, and 62.3 +/- 33.9 Bq kg(-1), respectively) are 3.5-8 times higher than the corresponding values in cultivated soils (0.147 +/- 0.054, 0.005 +/- 0.002, 0.047 +/- 0.021, 1.53 +/- 0.44, and 7.70 +/- 2.07 Bq kg(-1)). Radionuclide inventories and ratios are consistent with values reported by the United Nations Scientific Committee on the Effect of Atomic Radiation for this latitude (40-50 degrees N). This suggests that radiocontamination in this region is mainly due to atmospheric deposition of nuclear weapon test fallout. The vertical distribution of these radionuclides is also studied. The results show that, with the exception of 90Sr (68%), more than 90% of these radionuclides are contained in the first 20 cm of soil and that mobility follows the order 90Sr > 241Am > 239+240Pu, 238Pu > 137Cs. PMID- 10376544 TI - Resolving Nevada test site and global fallout plutonium in attic dust and soils using 137Cs/239+240Pu activity ratios. AB - A simple equation using only 137Cs/239+240Pu activity ratios was developed and evaluated as a means of resolving the plutonium in attic dust and soil from Nevada and Utah that came from Nevada Test Site fallout from that which came from global fallout. Applied to an historical data set of 137Cs and 239+240Pu activity concentrations in soils from Nevada and Utah, the activity ratio method gives results similar to the traditional 240Pu/239Pu isotope mass ratio method. Considering the difficulty and expense of determining the 240Pu/239Pu atom ratios, this activity ratio method is simpler, faster, and less costly, and may be useful for detecting and/or monitoring plutonium contamination in soils. Applied to samples of attic dust and soil collected from throughout southern Nevada and Utah during 1996 and 1997, it was found that all sites surveyed showed the presence of Nevada Test Site plutonium. Over 90% of the plutonium found in the samples from Beatty, Tonopah, and Queen City Summit, Nevada, can be attributed to the Nevada Test Site. PMID- 10376543 TI - Radioactive contamination of cistern waters along the Croatian coast of the Adriatic sea by 90Sr. AB - Measurements of radioactive contamination of water samples from cisterns collecting rainwater containing fission products from roofs and other surfaces have been carried out along the Croatian coast of the Adriatic sea since 1968. An exponential decline of radioactivity followed the nuclear moratorium. After the nuclear accident at Chernobyl, higher levels of 137Cs and 90Sr were detected again, with cistern waters being the only environmental samples in Croatia in which elevated 90Sr activities persisted for several years. For the pre-Chernobyl period, the observed mean residence time of 90Sr in cistern waters, estimated to be 6.2 +/- 1.9 y, was similar to that calculated for fallout. Contrary, for the post-Chernobyl time, observed 90Sr mean residence time was calculated to be considerably shorter, reflecting the tropospheric mean residence time. The annual dose for the critical adult population received from 90Sr and 137Cs by drinking cistern water was estimated to be very small, in the 1990's less than few microSv y(-1). PMID- 10376545 TI - A non-destructive method to determine the depth of radionuclides in materials in situ. AB - A non-destructive method based on in-situ gamma spectroscopy is developed to determine the depth of radiological contamination in media. An innovative algorithm, Gamma Penetration Depth Unfolding Algorithm (GPDUA), uses point kernel techniques to predict the depth of contamination based on the results of the uncollided peak information from the in-situ gamma spectroscopy. The GPDUA is designed and verified through extensive Monte Carlo simulations and validated through laboratory experiments. This innovative tool promises to be "better, faster, safer, and cheaper" than the current practice in decontamination and decommissioning. The method requires the a priori knowledge of the contaminant source distribution. The applicable radiological contaminants of interest are any isotopes that emit two or more gamma rays per disintegration or isotopes that emit a single gamma ray but have gamma-emitting progeny in secular equilibrium with its parent (e.g., 60Co, 235U, and 137Cs to name a few). The predicted depths from the GPDUA algorithm using Monte Carlo N-Particle Transport Code simulations and laboratory experiments using 60Co have consistently produced predicted depths within 20% of the actual or known depth. PMID- 10376546 TI - Stirring system for radioactive waste water storage tank. AB - A stirring system for 100-m(3) radioactive liquid waste tanks was constructed to unify radioactive concentrations in the tank. The stirring system is effective in certifying that the radioactive concentrations in the tanks are less than the legal limits before they are drained away as waste liquid. This system is composed of discharge units, pipe lines, and a controller. The performance of the system was assessed by comparing the calculated red ink and 32P concentrations with those monitored at six locations in the tanks. The concentration reached equilibrium after stirring 60 to 120 min with discharge units equipped with six fixed openings configured in differing directions. Residual chlorine in city water used for dilution occasionally bleached the red ink and reduced its concentration. The adsorption of 32P by slime on the walls of the tanks storing actual waste water lowered the equilibrium concentration. PMID- 10376547 TI - ICNIRP recommendation for limiting public exposure to 4-Hz-1-kHz electric and magnetic fields--need for peer review. PMID- 10376548 TI - A piece of my mind. Cold blue. PMID- 10376549 TI - A new source of resistance to HIV drugs. PMID- 10376550 TI - Why the rise in asthma? New insight, few answers. PMID- 10376551 TI - Respiratory research's reach. PMID- 10376552 TI - Survey says patients expect little physician help on sex. PMID- 10376553 TI - Treat arthritis earlier, better. PMID- 10376554 TI - From the Health Care Financing Administration. PMID- 10376555 TI - From the Centers for Disease Control and Prevention. Impact of arthritis and other rheumatic conditions on the health-care system--United States, 1997. PMID- 10376556 TI - From the Centers for Disease Control and Prevention. Update: influenza activity- United States and worldwide, 1998-99 season, and composition of the 1999-2000 influenza vaccine. PMID- 10376557 TI - From the Centers for Disease Control and Prevention. Outbreak of poliomyelitis- Angola, 1999. PMID- 10376558 TI - Autopsy rates and diagnosis. PMID- 10376559 TI - Autopsy rates and diagnosis. PMID- 10376560 TI - Autopsy rates and diagnosis. PMID- 10376561 TI - Autopsy rates and diagnosis. PMID- 10376562 TI - Autopsy rates and diagnosis. PMID- 10376563 TI - Autopsy rates and diagnosis. PMID- 10376564 TI - Autopsy rates and diagnosis. PMID- 10376565 TI - Autopsy rates and diagnosis. PMID- 10376566 TI - Autopsy rates and diagnosis. PMID- 10376567 TI - Autopsy rates and diagnosis. PMID- 10376568 TI - Interpretation of research on sexual abuse of boys. PMID- 10376569 TI - Interpretation of research on sexual abuse of boys. PMID- 10376570 TI - New perspectives in glaucoma. PMID- 10376572 TI - Trends in unintentional drowning: the role of alcohol and medical care. AB - CONTEXT: During the last few decades, mortality from drowning has decreased in the United States for unknown reasons. It has been hypothesized that this decline may be due to decreased use of alcohol in and around water or improved medical treatment after a submersion. OBJECTIVES: To estimate changes in unintentional mortality due to submersion, estimate trends in drownings attributable to alcohol use, and assess the role of medical care in these mortality trends. DESIGN: A 21 year longitudinal study of case findings, from January 1, 1975, through December 31,1995. SETTING AND PARTICIPANTS: All residents of King County, Washington, who died unintentionally from submersion and 284 persons hospitalized for submersion who survived. MAIN OUTCOME MEASURES: Changes in submersion-related mortality incidence over time, proportion of this mortality that could be attributed to alcohol use, changes over time in the case-fatality rate of treated patients, and estimate of deaths prevented in 1995 compared with projected estimates had there been no change in incidence since 1975. RESULTS: There were 539 deaths due to drowning in King County during 21 years. Mortality rates during this period declined by 59% (95% confidence interval [CI], -70% to -46%). The incidence of death attributable to alcohol use decreased by 81% (95% CI, -91% to -57%); this could account for 51% of deaths prevented in 1995. Among 249 comatose patients who received prehospital care, 205 died; the odds of survival decreased 40% over 21 years (P = .40). Among 101 comatose patients who were hospitalized, 63 died; the odds of survival decreased 29% (P = .75). The incidence of survival of comatose hospital patients decreased by 29% from 1975 to 1995 (95% CI, -78% to +125%). We found no evidence that trends in medical treatment prevented any deaths due to drowning in 1995. CONCLUSIONS: Drowning incidence in King County, Washington, declined because of a decrease in severe submersion episodes rather than an increase in success of medical interventions. Our data support the theory that less use of alcohol around water prevents some deaths. About half of the decrease was unexplained. PMID- 10376571 TI - The effect of raloxifene on risk of breast cancer in postmenopausal women: results from the MORE randomized trial. Multiple Outcomes of Raloxifene Evaluation. AB - CONTEXT: Raloxifene hydrochloride is a selective estrogen receptor modulator that has antiestrogenic effects on breast and endometrial tissue and estrogenic effects on bone, lipid metabolism, and blood clotting. OBJECTIVE: To determine whether women taking raloxifene have a lower risk of invasive breast cancer. DESIGN AND SETTING: The Multiple Outcomes of Raloxifene Evaluation (MORE), a multicenter, randomized, double-blind trial, in which women taking raloxifene or placebo were followed up for a median of 40 months (SD, 3 years), from 1994 through 1998, at 180 clinical centers composed of community settings and medical practices in 25 countries, mainly in the United States and Europe. PARTICIPANTS: A total of 7705 postmenopausal women, younger than 81 (mean age, 66.5) years, with osteoporosis, defined by the presence of vertebral fractures or a femoral neck or spine T-score of at least 2.5 SDs below the mean for young healthy women. Almost all participants (96%) were white. Women who had a history of breast cancer or who were taking estrogen were excluded. INTERVENTION: Raloxifene, 60 mg, 2 tablets daily; or raloxifene, 60 mg, 1 tablet daily and 1 placebo tablet; or 2 placebo tablets. MAIN OUTCOME MEASURES: New cases of breast cancer, confirmed by histopathology. Transvaginal ultrasonography was used to assess the endometrial effects of raloxifene in 1781 women. Deep vein thrombosis or pulmonary embolism were determined by chart review. RESULTS: Thirteen cases of breast cancer were confirmed among the 5129 women assigned to raloxifene vs 27 among the 2576 women assigned to placebo (relative risk [RR], 0.24; 95% confidence interval [CI], 0.13-0.44; P<.001). To prevent 1 case of breast cancer, 126 women would need to be treated. Raloxifene decreased the risk of estrogen receptor-positive breast cancer by 90% (RR, 0.10; 95% CI, 0.04-0.24), but not estrogen receptor-negative invasive breast cancer (RR, 0.88; 95% CI, 0.26-3.0). Raloxifene increased the risk of venous thromboembolic disease (RR, 3.1; 95% CI, 1.5-6.2), but did not increase the risk of endometrial cancer (RR, 0.8; 95% CI, 0.2-2.7). CONCLUSION: Among postmenopausal women with osteoporosis, the risk of invasive breast cancer was decreased by 76% during 3 years of treatment with raloxifene. PMID- 10376573 TI - Preoperative serum potassium levels and perioperative outcomes in cardiac surgery patients. Multicenter Study of Perioperative Ischemia Research Group. AB - CONTEXT: Although potassium is critical for normal electrophysiology, the association between abnormal preoperative serum potassium level and perioperative adverse events such as arrhythmias has not been examined rigorously. OBJECTIVE: To determine the prevalence of abnormal preoperative serum potassium levels and whether such abnormal levels are associated with adverse perioperative events. DESIGN AND SETTING: Prospective, observational, case-control study of data collected from 24 diverse US medical centers in a 2-year period from September 1, 1991, to September 1, 1993. PATIENTS: A total of 2402 patients (mean [SD] age, 65.1 [10.3] years; 24% female) undergoing elective coronary artery bypass grafting who were not enrolled in another protocol. The study population was identified using systematic sampling of every nth patient, in which n was based on expected total number of procedures at that center during the study period. MAIN OUTCOME MEASURES: Intraoperative and postoperative arrhythmias, the need for cardiopulmonary resuscitation (CPR), cardiac death, and death due to any cause prior to discharge, by preoperative serum potassium level. RESULTS: Perioperative arrhythmias occurred in 1290 (53.7%) of 2402 patients, with 238 patients (10.7%) having intraoperative arrhythmias, 329 (13.7%) having postoperative nonatrial arrhythmias, and 865 (36%) having postoperative atrial flutter or fibrillation. The incidence of adverse outcomes was 3.6% for death, 2.0% for cardiac death, and 3.5% for CPR. Serum potassium level less than 3.5 mmol/L was a predictor of serious perioperative arrhythmia (odds ratio [OR], 2.2; 95% confidence interval [CI], 1.2-4.0), intraoperative arrhythmia (OR, 2.0; 95% CI, 1.0-3.6), and postoperative atrial fibrillation/flutter (OR, 1.7; 95% CI, 1.0-2.7), and these relationships were unchanged after adjusting for confounders. The significant univariate association between increased need for CPR and serum potassium level less than 3.3 mmol/L (OR, 3.3; 95% CI, 1.2-9.5) and greater than 5.2 mmol/L (OR, 3.0; 95% CI, 1.1-8.7) became nonsignificant after adjusting for confounders. CONCLUSIONS: Perioperative arrhythmia and the need for CPR increased as preoperative serum potassium level decreased below 3.5 mmol/L. Although interventional trials are required to determine whether preoperative intervention mitigates these adverse associations, preoperative repletion is low cost and low risk, and our data suggest that screening and repletion be considered in patients scheduled for cardiac surgery. PMID- 10376574 TI - Carrier rates in the midwestern United States for GJB2 mutations causing inherited deafness. AB - CONTEXT: Mutations in the GJB2 gene are the most common known cause of inherited congenital severe-to-profound deafness. The carrier frequency of these mutations is not known. OBJECTIVES: To determine the carrier rate of deafness-causing mutations in GJB2 in the midwestern United States and the prevalence of these mutations in persons with congenital sensorineural hearing loss ranging in severity from moderate to profound, and to derive revised data for counseling purposes. DESIGN: Laboratory analysis, performed in 1998, of samples from probands with hearing loss for mutations in GJB2 using an allele-specific polymerase chain reaction assay, single-strand conformation polymorphism analysis, and direct sequencing. SETTING AND SUBJECTS: Fifty-two subjects younger than 19 years sequentially referred to a midwestern tertiary referral center for hearing loss or cochlear implantation, with moderate-to-profound congenital hearing loss of unknown cause, parental nonconsanguinity, and nonsyndromic deafness with hearing loss limited to a single generation; 560 control neonates were screened for the 35delG mutation. MAIN OUTCOME MEASURE: Prevalence of mutations in the GJB2 gene by congenital deafness status. RESULTS: Of 52 sequential probands referred for congenital sensorineural hearing loss, 22 (42%) were found to have GJB2 mutations. The 35delG mutation was identified in 29 of the 41 mutant alleles. Of probands' sibs, all homozygotes and compound heterozygotes had deafness. Fourteen of 560 controls were 35delG heterozygotes, for a carrier rate expressed as a mean (SE) of 2.5% (0.66%). The carrier rate for all recessive deafness-causing GJB2 mutations was determined to be 3.01% (probable range, 2.54%-3.56%). Calculated sensitivity and specificity for a screening test based on 35delG mutation alone were 96.9% and 97.4%, respectively, and observed values were 94% and 97%, respectively. CONCLUSIONS: Our data suggest that mutations in GJB2 are the leading cause of moderate-to-profound congenital inherited deafness in the midwestern United States. Screening of the GJB2 mutation can be offered to individuals with congenital deafness with high sensitivity and specificity by screening only for the 35delG mutation. A positive finding should establish an etiologic diagnosis and affect genetic counseling. PMID- 10376575 TI - Proportion of cystic fibrosis gene mutations not detected by routine testing in men with obstructive azoospermia. AB - CONTEXT: Infertile men with obstructive azoospermia may have mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, many of which are rare in classic cystic fibrosis and not evaluated in most routine mutation screening. OBJECTIVE: To assess how often CFTR mutations or sequence alterations undetected by routine screening are detected with more extensive screening in obstructive azoospermia. DESIGN: Routine screening for the 31 most common CFTR mutations associated with the CF phenotype in white populations, testing for the 5-thymidine variant of the polythymidine tract of intron 8 (IVS8-5T) by allele specific oligonucleotide hybridization, and screening of all exons through multiplex heteroduplex shift analysis followed by direct DNA sequencing. SETTING: Male infertility clinic of a Canadian university-affiliated hospital. SUBJECTS: Of 198 men with obstructive (n = 149) or nonobstructive (n = 49; control group) azoospermia, 64 had congenital bilateral absence of the vas deferens (CBAVD), 10 had congenital unilateral absence of the vas deferens (CUAVD), and 75 had epididymal obstruction (56/75 were idiopathic). MAIN OUTCOME MEASURE: Frequency of mutations found by routine and nonroutine tests in men with obstructive vs nonobstructive azoospermia. RESULTS: Frequency of mutations and the IVS8-5T variant in the nonobstructive azoospermia group (controls) (2% and 5.1% allele frequency, respectively) did not differ significantly from that in the general population (2% and 5.2%, respectively). In the CBAVD group, 72 mutations were found by DNA sequencing and IVS8-5T testing (47 and 25, respectively; P<.001 and P = .002 vs controls) vs 39 by the routine panel (P<.001 vs controls). In the idiopathic epididymal obstruction group, 24 mutations were found by DNA sequencing and IVS8-5T testing (12 each; P=.01 and P=.14 vs controls) vs 5 by the routine panel (P=.33 vs controls). In the CUAVD group, 2 mutations were found by routine testing (P=.07 vs controls) vs 4 (2 each, respectively; P=.07 and P=.40 vs controls) by DNA sequencing and IVS8-5T testing. The routine panel did not identify 33 (46%) of 72, 2 (50%) of 4, and 19 (79%) of 24 detectable CFTR mutations and IVS8-5T in the CBAVD, CUAVD, and idiopathic epididymal obstruction groups, respectively. CONCLUSIONS: Routine testing for CFTR mutations may miss mild or rare gene alterations. The barrier to conception for men with obstructive infertility has been overcome by assisted reproductive technologies, thus raising the concern of iatrogenically transmitting pathogenic CFTR mutations to the progeny. PMID- 10376576 TI - Reticulocyte hemoglobin content to diagnose iron deficiency in children. AB - CONTEXT: Early identification of iron deficiency in children is essential to prevent the damaging long-term consequences of this disease. However, it is not clear which indices should be included in a diagnostic panel for iron deficiency and iron deficiency anemia in children. OBJECTIVE: To develop an effective approach for the diagnosis of iron deficiency and iron deficiency anemia in young children. DESIGN AND SETTING: Retrospective laboratory analysis, carried out over 7 weeks in 1996, using blood samples ordered by pediatricians and sent to a large metropolitan hospital for analysis. PATIENTS: A total of 210 children (mean [SD] age, 2.9 [2.0] years; 120 were male) who had a lead screening test (complete blood cell count and plasma lead level) ordered by a primary care pediatrician. MAIN OUTCOME MEASURES: Levels of hemoglobin (Hb), iron, transferrin, transferrin saturation (Tfsat), ferritin, and circulating transferrin receptor and reticulocyte Hb content (CHr) among patients with and without iron deficiency, defined as Tfsat of less than 20%, and iron deficiency anemia, defined as Tfsat of less than 20% and Hb level of less than 110 g/L. RESULTS: Of the 210 subjects, 43 (20.5%) were iron deficient; 24 of these had iron deficiency anemia. Reticulocyte Hb content and Hb levels were the only significant predictors of iron deficiency (likelihood ratio test [LRT] = 15.96; P<.001 for CHr, and LRT = 6.59; P = .01 for Hb), and CHr was the only significant multivariate predictor of iron deficiency anemia (LRT = 30.43; P<.001). Plasma ferritin level had no predictive value (P = .97). Subjects with CHr of less than 26 pg (optimal cutoff value based on sensitivity/specificity analysis) had lower Hb level, mean corpuscular volume, mean corpuscular Hb level, serum iron level, and Tfsat, and increased red blood cell distribution width vs those with CHr of 26 pg or more (P<.001 for all). CONCLUSIONS: Reticulocyte Hb content level was the strongest predictor of iron deficiency and iron deficiency anemia in children. It holds promise as an alternative to biochemical iron studies in diagnosis. PMID- 10376578 TI - Editorial governance of the Journal of the American Medical Association: a report. PMID- 10376577 TI - The rational clinical examination. Does this patient have aortic regurgitation? AB - OBJECTIVE: To review evidence as to the precision and accuracy of clinical examination for aortic regurgitation (AR). METHODS: We conducted a structured MEDLINE search of English-language articles (January 1966-July 1997), manually reviewed all reference lists of potentially relevant articles, and contacted authors of relevant studies for additional information. Each study (n = 16) was independently reviewed by both authors and graded for methodological quality. RESULTS: Most studies assessed cardiologists as examiners. Cardiologists' precision for detecting diastolic murmurs was moderate using audiotapes (kappa = 0.51) and was good in the clinical setting (simple agreement, 94%). The most useful finding for ruling in AR is the presence of an early diastolic murmur (positive likelihood ratio [LR], 8.8-32.0 [95% confidence interval [CI], 2.8-32 to 16-63] for detecting mild or greater AR and 4.0-8.3 [95% CI, 2.5-6.9 to 6.2 11] for detecting moderate or greater AR) (2 grade A studies). The most useful finding for ruling out AR is the absence of early diastolic murmur (negative LR, 0.2-0.3 [95% CI, 0.1-0.3 to 0.2-0.4) for mild or greater AR and 0.1 [95% CI, 0.0 0.3] for moderate or greater AR) (2 grade A studies). Except for a test evaluating the response to transient arterial occlusion (positive LR, 8.4 [95% CI, 1.3-81.0]; negative LR, 0.3 [95% CI, 0.1-0.8]), most signs display poor sensitivity and specificity for AR. CONCLUSION: Clinical examination by cardiologists is accurate for detecting AR, but not enough is known about the examinations of less-expert clinicians. PMID- 10376579 TI - Encouraging news from the SERM frontier. Selective estrogen receptor modulator. PMID- 10376580 TI - Declines in drowning: exploring the epidemiology of favorable trends. PMID- 10376582 TI - JAMA Patient Page: water safety. PMID- 10376581 TI - Choosing the best strategy to prevent childhood iron deficiency. PMID- 10376583 TI - Help! The data are coming. PMID- 10376584 TI - Biotech panel set up in US may help allay public fears. PMID- 10376585 TI - Varmus relaunches NIH degree proposal. PMID- 10376586 TI - US jury split over hormone patent case... PMID- 10376587 TI - as Seeburg faces misconduct inquiry. PMID- 10376588 TI - It's sink or swim as a tidal wave of data approaches. PMID- 10376589 TI - Longevity--does family size matter? PMID- 10376591 TI - Looking out for memory T cells. PMID- 10376590 TI - Antibiotic resistance. A vancomycin surprise. PMID- 10376592 TI - Hiding messages in DNA microdots. PMID- 10376593 TI - Marine viruses and their biogeochemical and ecological effects. AB - Viruses are the most common biological agents in the sea, typically numbering ten billion per litre. They probably infect all organisms, can undergo rapid decay and replenishment, and influence many biogeochemical and ecological processes, including nutrient cycling, system respiration, particle size-distributions and sinking rates, bacterial and algal biodiversity and species distributions, algal bloom control, dimethyl sulphide formation and genetic transfer. Newly developed fluorescence and molecular techniques leave the field poised to make significant advances towards evaluating and quantifying such effects. PMID- 10376594 TI - Structural basis of procaspase-9 recruitment by the apoptotic protease-activating factor 1. AB - Caspase-9-mediated apoptosis (programmed cell death) plays a central role in the development and homeostasis of all multicellular organisms. Mature caspase-9 is derived from its procaspase precursor as a result of recruitment by the activating factor Apaf-1. The crystal structures of the caspase-recruitment domain of Apaf-1 by itself and in complex with the prodomain of procaspase-9 have been determined at 1.6 and 2.5 A resolution, respectively. These structures and other evidence reveal that each molecule of Apaf-1 interacts with a molecule of procaspase-9 through two highly charged and complementary surfaces formed by non conserved residues; these surfaces determine recognition specificity through networks of intermolecular hydrogen bonds and van der Waals interactions. Mutation of the important interface residues in procaspase-9 or Apaf-1 prevents or reduces activation of procaspase-9 in a cell-free system. Wild-type, but not mutant, prodomains of caspase-9 completely inhibit catalytic processing of procaspase-9. Furthermore, analysis of homologues from Caenorhabditis elegans indicates that recruitment of CED-3 by CED-4 is probably mediated by the same set of conserved structural motifs, with a corresponding change in the specificity determining residues. PMID- 10376595 TI - Dynamics of individual flexible polymers in a shear flow. AB - Polymer dynamics are of central importance in materials science, mechanical engineering, biology and medicine. The dynamics of macromolecular solutions and melts in shear flow are typically studied using bulk experimental methods such as light and neutron scattering and birefringence. But the effect of shear on the conformation and dynamics of individual polymers is still not well understood. Here we describe observations of the real-time dynamics of individual, flexible polymers (fluorescently labelled DNA molecules) under a shear flow. The sheared polymers exhibit many types of extended conformation with an overall orientation ranging from parallel to perpendicular with respect to the flow direction. For shear rates much smaller than the inverse of the relaxation time of the molecule, the relative populations of these two main types of conformation are controlled by the rate of the shear flow. These results question the adequacy of assumptions made in standard models of polymer dynamics. PMID- 10376596 TI - Tuning bilayer twist using chiral counterions. AB - From seashells to DNA, chirality is expressed at every level of biological structures. In self-assembled structures it may emerge cooperatively from chirality at the molecular scale. Amphiphilic molecules, for example, can form a variety of aggregates and mesophases that express the chirality of their constituent molecules at a supramolecular scale of micrometres. Quantitative prediction of the large-scale chirality based on that at the molecular scale remains a largely unsolved problem. Furthermore, experimental control over the expression of chirality at the supramolecular level is difficult to achieve: mixing of different enantiomers usually results in phase separation. Here we present an experimental and theoretical description of a system in which chirality can be varied continuously and controllably ('tuned') in micrometre scale structures. We observe the formation of twisted ribbons consisting of bilayers of gemini surfactants (two surfactant molecules covalently linked at their charged head groups). We find that the degree of twist and the pitch of the ribbons can be tuned by the introduction of opposite-handed chiral counterions in various proportions. This degree of control might be of practical value; for example, in the use of the helical structures as templates for helical crystallization of macromolecules. PMID- 10376597 TI - Feature-based attention influences motion processing gain in macaque visual cortex. AB - Changes in neural responses based on spatial attention have been demonstrated in many areas of visual cortex, indicating that the neural correlate of attention is an enhanced response to stimuli at an attended location and reduced responses to stimuli elsewhere. Here we demonstrate non-spatial, feature-based attentional modulation of visual motion processing, and show that attention increases the gain of direction-selective neurons in visual cortical area MT without narrowing the direction-tuning curves. These findings place important constraints on the neural mechanisms of attention and we propose to unify the effects of spatial location, direction of motion and other features of the attended stimuli in a 'feature similarity gain model' of attention. PMID- 10376598 TI - Assignment of circadian function for the Neurospora clock gene frequency. AB - Circadian clocks consist of three elements: entrainment pathways (inputs), the mechanism generating the rhythmicity (oscillator), and the output pathways that control the circadian rhythms. It is difficult to assign molecular clock components to any one of these elements. Experiments show that inputs can be circadianly regulated and outputs can feed back on the oscillator. Mathematical simulations indicate that under- or overexpression of a gene product can result in arrhythmicity, whether the protein is part of the oscillator or substantially part of a rhythmically expressed input pathway. To distinguish between these two possibilities, we used traditional circadian entrainment protocols on a genetic model system, Neurospora crassa. PMID- 10376599 TI - Control of organ shape by a secreted metalloprotease in the nematode Caenorhabditis elegans. AB - The molecular controls governing organ shape are poorly understood. In the nematode Caenorhabditis elegans, the gonad acquires a U-shape by the directed migration of a specialized 'leader' cell, which is located at the tip of the growing gonadal 'arm'. The gon-1 gene is essential for gonadal morphogenesis: in gon-1 mutants, no arm elongation occurs and somatic gonadal structures are severely malformed. Here we report that gon-1 encodes a secreted protein with a metalloprotease domain and multiple thrombospondin type-1-like repeats. This motif architecture is typical of a small family of genes that include bovine procollagen I N-protease (P1NP), which cleaves collagen, and murine ADAMTS-1, the expression of which correlates with tumour cell progression. We find that gon-1 is expressed in two sites, leader cells and muscle, and that expression in each site has a unique role in forming the gonad. We speculate that GON-1 controls morphogenesis by remodelling basement membranes and that regulation of its activity is crucial for achieving organ shape. PMID- 10376600 TI - Emergence of vancomycin tolerance in Streptococcus pneumoniae. AB - Streptococcus pneumoniae, the pneumococcus, is the most common cause of sepsis and meningitis. Multiple-antibiotic-resistant strains are widespread, and vancomycin is the antibiotic of last resort. Emergence of vancomycin resistance in this community-acquired bacterium would be catastrophic. Antibiotic tolerance, the ability of bacteria to survive but not grow in the presence of antibiotics, is a precursor phenotype to resistance. Here we show that loss of function of the VncS histidine kinase of a two-component sensor-regulator system in S. pneumoniae produced tolerance to vancomycin and other classes of antibiotic. Bacterial two component systems monitor environmental parameters through a sensor histidine kinase/phosphatase, which phosphorylates/dephosphorylates a response regulator that in turn mediates changes in gene expression. These results indicate that signal transduction is critical for the bactericidal activity of antibiotics. Experimental meningitis caused by the vncS mutant failed to respond to vancomycin. Clinical isolates tolerant to vancomycin were identified and DNA sequencing revealed nucleotide alterations in vncS. We conclude that broad antibiotic tolerance of S. pneumoniae has emerged in the community by a molecular mechanism that eliminates sensitivity to the current cornerstone of therapy, vancomycin. PMID- 10376601 TI - Modelling T-cell memory by genetic marking of memory T cells in vivo. AB - Immunological memory is the ability of the immune system to respond with enhanced vigour to pathogens that have been encountered in the past. Following infection or immunization, most effector T cells undergo apoptotic cell death, but a small fraction of these cells, proportional to the early antigen load and initial clonal burst size, persist in the host as a stable pool of memory T cells. The existence of immunological memory has been recognized for over 2,000 years, but our understanding of this phenomenon is limited, primarily because memory lymphocytes cannot be unequivocally identified as they lack specific, permanent markers. Here we have developed a transgenic mouse model system whereby memory T cells and their precursors can be irreversibly marked with a reporter gene and thus can be unambiguously identified. Adoptive transfer of marked CD8+ T cells specific for lymphocytic choriomeningitis virus protected naive recipients following viral challenge, demonstrating that we have marked memory T cells. We also show that cytotoxic effector lymphocytes that develop into memory T cells can be identified in the primary response. PMID- 10376603 TI - Activation of nitric oxide synthase in endothelial cells by Akt-dependent phosphorylation. AB - Nitric oxide (NO) produced by the endothelial NO synthase (eNOS) is a fundamental determinant of cardiovascular homesotasis: it regulates systemic blood pressure, vascular remodelling and angiogenesis. Physiologically, the most important stimulus for the continuous formation of NO is the viscous drag (shear stress) generated by the streaming blood on the endothelial layer. Although shear-stress mediated phosphorylation of eNOS is thought to regulate enzyme activity, the mechanism of activation of eNOS is not yet known. Here we demonstrate that the serine/threonine protein kinase Akt/PKB mediates the activation of eNOS, leading to increased NO production. Inhibition of the phosphatidylinositol-3-OH kinase/Akt pathway or mutation of the Akt site on eNOS protein (at serine 1177) attenuates the serine phosphorylation and prevents the activation of eNOS. Mimicking the phosphorylation of Ser 1177 directly enhances enzyme activity and alters the sensitivity of the enzyme to Ca2+, rendering its activity maximal at sub-physiological concentrations of Ca2+. Thus, phosphorylation of eNOS by Akt represents a novel Ca2+-independent regulatory mechanism for activation of eNOS. PMID- 10376602 TI - Regulation of endothelium-derived nitric oxide production by the protein kinase Akt. AB - Endothelial nitric oxide synthase (eNOS) is the nitric oxide synthase isoform responsible for maintaining systemic blood pressure, vascular remodelling and angiogenesis. eNOS is phosphorylated in response to various forms of cellular stimulation, but the role of phosphorylation in the regulation of nitric oxide (NO) production and the kinase(s) responsible are not known. Here we show that the serine/threonine protein kinase Akt (protein kinase B) can directly phosphorylate eNOS on serine 1179 and activate the enzyme, leading to NO production, whereas mutant eNOS (S1179A) is resistant to phosphorylation and activation by Akt. Moreover, using adenovirus-mediated gene transfer, activated Akt increases basal NO release from endothelial cells, and activation-deficient Akt attenuates NO production stimulated by vascular endothelial growth factor. Thus, eNOS is a newly described Akt substrate linking signal transduction by Akt to the release of the gaseous second messenger NO. PMID- 10376604 TI - Enhancement of TBP binding by activators and general transcription factors. AB - Eukaryotic transcriptional activators function, at least in part, by promoting assembly of the preinitiation complex, which comprises RNA polymerase II and its general transcription factors (GTFs). Activator-mediated stimulation of the assembly of the preinitiation complex has been studied in vitro but has been relatively refractory to in vivo analysis. Here we use a DNA crosslinking/immunoprecipitation assay to study in living cells the first step in the assembly of the preinitiation complex, the interaction between the TATA-box binding protein (TBP) and its binding site, the TATA box. Analysis of a variety of endogenous yeast genes, and of a series of activators of differing strength, reveals a general correlation between TBP binding and transcriptional activity. Using mutant yeast strains, we show that Mot1 prevents the binding of TBP to inactive promoters and that activator-mediated stimulation of TBP binding requires additional GTFs, including TFIIB and Srb4. Taken together, our results indicate that TBP binding in vivo is stringently controlled, and that the ability of activators to stimulate this step in the assembly of the preinitiation complex is a highly cooperative process involving multiple transcription factors. PMID- 10376606 TI - NIH E-biomed proposal: a welcome jolt. PMID- 10376605 TI - Binding of TBP to promoters in vivo is stimulated by activators and requires Pol II holoenzyme. AB - In eukaryotes, transcriptional activators have been proposed to function by recruiting the RNA polymerase II (Pol II) machinery, by altering the conformation of this machinery, or by affecting steps after initiation, but the evidence is not definitive. Genomic footprinting of yeast TATA-box elements reveals activator dependent alterations of chromatin structure and activator-independent protection, but little is known about the association of specific components of the Pol II machinery with promoters in vivo. Here we analyse TATA-box-binding protein (TBP) occupancy of 30 yeast promoters in vivo. We find that TBP association with promoters is stimulated by activators and inhibited by the Cyc8 Tup1 repressor, and that transcriptional activity correlates strongly with the degree of TBP occupancy. In a small subset of promoters, TBP occupancy is higher than expected when gene activity is low, and the activator-dependent increase is modest. TBP association depends on the Pol II holoenzyme component Srb4, but not on the Kin28 subunit of the transcription factor TFIIH, even though both proteins are generally required for transcription. Thus in yeast cells, TBP association with promoters occurs in concert with the Pol II holoenzyme, activator-dependent recruitment of the Pol II machinery occurs at the vast majority of promoters, and Kin28 acts after the initial recruitment. PMID- 10376607 TI - Tamoxifen hits the target in situ. PMID- 10376608 TI - Negative association between MMR and autism. PMID- 10376609 TI - Benefit of beta-blockers for heart failure: proven in 1999. PMID- 10376611 TI - Disability in older people: a mass problem requiring mass solutions. PMID- 10376610 TI - Relevance of AIDS treatment with two nucleoside analogues alone. PMID- 10376612 TI - Japan's loss of leadership role in access to drug data. PMID- 10376613 TI - Tamoxifen in treatment of intraductal breast cancer: National Surgical Adjuvant Breast and Bowel Project B-24 randomised controlled trial. AB - BACKGROUND: We have shown previously that lumpectomy with radiation therapy was more effective than lumpectomy alone for the treatment of ductal carcinoma in situ (DCIS). We did a double-blind randomised controlled trial to find out whether lumpectomy, radiation therapy, and tamoxifen was of more benefit than lumpectomy and radiation therapy alone for DCIS. METHODS: 1804 women with DCIS, including those whose resected sample margins were involved with tumour, were randomly assigned lumpectomy, radiation therapy (50 Gy), and placebo (n=902), or lumpectomy, radiation therapy, and tamoxifen (20 mg daily for 5 years, n=902). Median follow-up was 74 months (range 57-93). We compared annual event rates and cumulative probability of invasive or non-invasive ipsilateral and contralateral tumours over 5 years. FINDINGS: Women in the tamoxifen group had fewer breast cancer events at 5 years than did those on placebo (8.2 vs 13.4%, p=0.0009). The cumulative incidence of all invasive breast-cancer events in the tamoxifen group was 4.1% at 5 years: 2.1% in the ipsilateral breast, 1.8% in the contralateral breast, and 0.2% at regional or distant sites. The risk of ipsilateral-breast cancer was lower in the tamoxifen group even when sample margins contained tumour and when DCIS was associated with comedonecrosis. INTERPRETATION: The combination of lumpectomy, radiation therapy, and tamoxifen was effective in the prevention of invasive cancer. PMID- 10376614 TI - Effect of metoprolol CR/XL in chronic heart failure: Metoprolol CR/XL Randomised Intervention Trial in Congestive Heart Failure (MERIT-HF) AB - BACKGROUND: Metoprolol can improve haemodynamics in chronic heart failure, but survival benefit has not been proven. We investigated whether metoprolol controlled release/extended release (CR/XL) once daily, in addition to standard therapy, would lower mortality in patients with decreased ejection fraction and symptoms of heart failure. METHODS: We enrolled 3991 patients with chronic heart failure in New York Heart Association (NYHA) functional class II-IV and with ejection fraction of 0.40 or less, stabilised with optimum standard therapy, in a double-blind randomised controlled study. Randomisation was preceded by a 2-week single-blind placebo run-in period. 1990 patients were randomly assigned metoprolol CR/XL 12.5 mg (NYHA III-IV) or 25.0 mg once daily (NYHA II) and 2001 were assigned placebo. The target dose was 200 mg once daily and doses were up titrated over 8 weeks. Our primary endpoint was all-cause mortality, analysed by intention to treat. FINDINGS: The study was stopped early on the recommendation of the independent safety committee. Mean follow-up time was 1 year. All-cause mortality was lower in the metoprolol CR/XL group than in the placebo group (145 [7.2%, per patient-year of follow-up]) vs 217 deaths [11.0%], relative risk 0.66 [95% CI 0.53-0.81]; p=0.00009 or adjusted for interim analyses p=0.0062). There were fewer sudden deaths in the metoprolol CR/XL group than in the placebo group (79 vs 132, 0.59 [0.45-0.78]; p=0.0002) and deaths from worsening heart failure (30 vs 58, 0.51 [0.33-0.79]; p=0.0023). INTERPRETATION: Metoprolol CR/XL once daily in addition to optimum standard therapy improved survival. The drug was well tolerated. PMID- 10376615 TI - Optimisation of antihypertensive treatment by crossover rotation of four major classes. AB - BACKGROUND: Most comparisons of antihypertensive drugs are undertaken in parallel groups. We undertook a crossover rotation of the four main classes of antihypertensive drugs, in untreated young hypertensive patients, to assess the response rate with monotherapy achieved by a systematic rotation. METHODS: 56 patients, mean blood pressure 161/98 mm Hg, entered the rotation, of whom 36 received all four monthly cycles of treatment with an angiotensin-converting enzyme (ACE) inhibitor (A), beta-blocker (B), calcium-channel blocker (C), and diuretic (D). Each patient's best drug was then repeated to assess repeatability. Two measures of individual variability in response were used. First, the value of rotation was measured by the increased proportion of patients reaching target blood pressure on their best drug versus their first drug. Second, we assessed whether the responses to each drug were correlated with each other. FINDINGS: Significant variability in response was found. 20 of the 41 patients reaching target blood pressure (< or =140/90 mm Hg) failed to achieve this target on their first drug. Rotation increased from 22/56 (39%) to 41/56 (73%) the success of monotherapy (p=0.0001); in half the patients, blood-pressure on the best treatment was 135/85 mm Hg or less. There were significant correlations between the blood pressure responses to A and B (r=0.5, p<0.01), and C and D (r=0.6, p<0.001), but not between the other four pairings of treatments. The responses to the AB pair were, on average, at least 50% higher than those to the CD pair; this difference was highly significant by multivariate repeated-measures ANOVA. INTERPRETATION: There is a marked variability in hypertensive patients' response to different antihypertensive drugs. The basis may be underlying variability in types of essential hypertension. Optimisation of treatment requires systematic rotation through several therapies; however, an "AB/CD" rule is proposed in which one of each of the two pairs of treatments is initially selected to abbreviate the rotation in routine practice. PMID- 10376616 TI - Zidovudine, didanosine, and zalcitabine in the treatment of HIV infection: meta analyses of the randomised evidence. HIV Trialists' Collaborative Group. AB - BACKGROUND: To assess the effects of zidovudine, didanosine, and zalcitabine on HIV disease progression and survival, we undertook meta-analyses of individual patient data and tabular data from all randomised trials that compared these agents. METHODS: Individual patient data were available for 7722 participants without AIDS in the nine randomised trials of immediate versus deferred zidovudine, and 7700 participants with or without AIDS in the six trials comparing zidovudine plus didanosine, zidovudine plus zalcitabine, or zidovudine alone. The main outcomes were mortality and disease progression (new AIDS defining event or death before any such event). FINDINGS: In the comparison of immediate versus deferred zidovudine, during a median follow-up of 50 months, 1908 individuals progressed, of whom 1351 died. In the deferred group, 61% started antiretroviral therapy (median time to therapy 28 months, which was zidovudine monotherapy in 94%). During the first year of follow-up, immediate zidovudine halved the rate of disease progression (p<0.0001), increasing the probability of AIDS-free survival at 1 year from 96% to 98%. This early delay did not persist: after 6 years, AIDS-free survival was 54% in both groups. At no time was there any difference in overall survival, which at 6 years was 64% with immediate and 65% with deferred zidovudine (rate ratio 1.04 [95% CI 0.94-1.15]). In the comparison of zidovudine plus didanosine or zalcitabine versus zidovudine alone, during a median follow-up of 29 months, 2904 individuals progressed, of whom 1850 died. The addition of didanosine to zidovudine delayed both progression (rate ratio 0.74 [0.67-0.82], p<0.0001) and death (0.72 [0.64-0.82], p<0.0001). Similarly, the addition of zalcitabine to zidovudine also delayed progression (0.86 [0.78-0.94], p=0.001) and death (0.87 [0.77-0.98], p=0.02). After 3 years, the estimated percentages alive and without a new AIDS event were 53% for zidovudine plus didanosine, 49% for zidovudine plus zalcitabine, and 44% for zidovudine alone; the percentages alive were 68%, 63%, and 59%, respectively. Five of the six trials involved randomised comparisons of zidovudine plus didanosine versus zidovudine plus zalcitabine: in these, the zidovudine plus didanosine regimen had greater effects on disease progression (p=0.004) and death (p=0.009). INTERPRETATION: Although immediate use of zidovudine halved disease progression during the first year, this effect was not sustained, and there was no improvement in survival in the short or long term. However, the use of didanosine and, to a lesser extent, zalcitabine delayed both disease progression and death, at least when added to zidovudine. The comparative effects of these different nucleoside analogues on long-term survival should inform the choice of which to combine with other types of drug, such as protease inhibitors. PMID- 10376617 TI - Autism and measles, mumps, and rubella vaccine: no epidemiological evidence for a causal association. AB - BACKGROUND: We undertook an epidemiological study to investigate whether measles, mumps, and rubella (MMR) vaccine may be causally associated with autism. METHODS: Children with autism born since 1979 were identified from special needs/disability registers and special schools in eight North Thames health districts, UK. Information from clinical records was linked to immunisation data held on the child health computing system. We looked for evidence of a change in trend in incidence or age at diagnosis associated with the introduction of MMR vaccination to the UK in 1988. Clustering of onsets within defined postvaccination periods was investigated by the case-series method. FINDINGS: We identified 498 cases of autism (261 of core autism, 166 of atypical autism, and 71 of Asperger's syndrome). In 293 cases the diagnosis could be confirmed by the criteria of the International Classification of Diseases, tenth revision (ICD10: 214 [82%] core autism, 52 [31%] atypical autism, 27 [38%] Asperger's syndrome). There was a steady increase in cases by year of birth with no sudden "step-up" or change in the trend line after the introduction of MMR vaccination. There was no difference in age at diagnosis between the cases vaccinated before or after 18 months of age and those never vaccinated. There was no temporal association between onset of autism within 1 or 2 years after vaccination with MMR (relative incidence compared with control period 0.94 [95% CI 0.60-1.47] and 1.09 [0.79 1.52]). Developmental regression was not clustered in the months after vaccination (relative incidence within 2 months and 4 months after MMR vaccination 0.92 [0.38-2.21] and 1.00 [0.52-1.95]). No significant temporal clustering for age at onset of parental concern was seen for cases of core autism or atypical autism with the exception of a single interval within 6 months of MMR vaccination. This appeared to be an artifact related to the difficulty of defining precisely the onset of symptoms in this disorder. INTERPRETATION: Our analyses do not support a causal association between MMR vaccine and autism. If such an association occurs, it is so rare that it could not be identified in this large regional sample. PMID- 10376618 TI - Decline in total serum IgE after treatment for tuberculosis. AB - BACKGROUND: Infection with Mycobacterium tuberculosis induces a type-1 immune response, whereas intestinal parasites elicit a type-2 response. Given that type 1 and type-2 responses inhibit each other, we investigated if M tuberculosis downregulates serum IgE, a marker of a type-2 response. METHODS: A prospective study was done in the Western Cape Province of South Africa, where tuberculosis and intestinal-parasite infection are common. Total serum IgE was determined for 37 controls and for 33 adolescent patients at presentation with tuberculosis and after successful completion of treatment. IgE specific for ascaris and allergens were measured in a subset of these individuals. Mantoux skin tests were done on 35 controls and on 31 patients at diagnosis. FINDINGS: Total IgE concentrations were high in controls (mean 313 kU/L) and in patients before treatment (mean 457 kU/L, p=0.085) and declined in all patients following successful treatment (mean 175 kU/L, p<0.0001). Posttreatment IgE concentrations did not differ from concentrations in controls. Ascaris-specific IgE was lower in controls (mean 1.73 kU/L) than in patients before treatment (4.62 kU/L, p=0.023) and was 2.39 kU/L in patients after treatment (p=0.0625). Tuberculin induration correlated inversely with IgE in patients but not in controls. INTERPRETATION: Infection with M tuberculosis as such is not incompatible with a prominent IgE response. IgE concentrations decreased after successful treatment of tuberculosis, showing that IgE concentrations in human beings can be downregulated under these circumstances, presumably due to enhancement of a type-1 response. PMID- 10376619 TI - A train driver with painful legs. PMID- 10376620 TI - Rehabilitation of hemiparesis after stroke with a mirror. PMID- 10376621 TI - Early recanalisation in acute ischaemic stroke saves tissue at risk defined by MRI. PMID- 10376622 TI - Aplastic anaemia in donor cells 14 years after bone-marrow transplant. PMID- 10376623 TI - Reducing incidence of headache after lumbar puncture and intrathecal cytotoxics. PMID- 10376624 TI - Isolation of enterohaemorrhagic Escherichia coli from municipal sewage. PMID- 10376625 TI - Pneumocystis carinii genotypes and severity of pneumonia. PMID- 10376626 TI - Intake of trans fatty acids and prevalence of childhood asthma and allergies in Europe. ISAAC Steering Committee. PMID- 10376627 TI - Clozapine in drug-induced psychosis in Parkinson's disease. The French Clozapine Parkinson Study Group. PMID- 10376628 TI - Topical cidofovir for severe molluscum contagiosum. PMID- 10376629 TI - Computers--a sympathetic shoulder to cry on. PMID- 10376630 TI - Health and wealth in the United Arab Emirates. PMID- 10376631 TI - European dioxin-contaminated food crisis grows and grows. PMID- 10376632 TI - Acute pain. AB - Postanaesthesia care units used to echo with cries of patients in pain after general anaesthesia. Each as-needed dose of analgesia was given only after permission of the surgeon or anaesthesiologist. Once conscious, patients were required to request each subsequent analgesic dose until hospital discharge. Not surprisingly, nearly half the patients who have an operation experience moderate to severe pain after surgery. Acute pain control has advanced dramatically and is now a field with dedicated texts, journals, and research. Despite improved surgical techniques that have transformed many operations into same-day procedures, inadequately controlled pain may still extend the length of hospital stay and predispose to expensive, time-consuming complications such as pneumonia. Recognition of economic and humanitarian benefits of pain control has prompted worldwide attention from professional groups, insurers, and governments. This paper describes the process of acute pain and measures to control it with drugs or non-pharmacological interventions. Even brief intervals of acute pain can induce long-term neuronal remodelling and sensitisation ("plasticity"), chronic pain, and lasting psychologial distress. Hence, acute pain and other types of pain (cancer-related or chronic) that are classified as distinct actually have many similarities. PMID- 10376633 TI - Disarming life's invisible enemies: Mikhail Bulgakov's A Country Doctor's Notebook. PMID- 10376634 TI - Analysis of HIV-1 clinical trials: statistical magic? The AVANTI Steering Committee. PMID- 10376635 TI - Physicians for Human Rights raise concerns for ethnic Albanians in refugee camps and those left inside Kosovo. PMID- 10376636 TI - Health and human rights of the East Timorese. PMID- 10376637 TI - Cholera in Madagascar. PMID- 10376638 TI - Cholera epidemiology. PMID- 10376639 TI - Psychiatric dysfunction and dizziness. PMID- 10376640 TI - Psychiatric dysfunction and dizziness. PMID- 10376641 TI - Statins and monocytes. PMID- 10376642 TI - Child obesity and body-mass index. PMID- 10376643 TI - Mammography in Asian patients with BRCA1 mutations. PMID- 10376644 TI - Mammography in Asian patients with BRCA1 mutations. PMID- 10376645 TI - Interaction between sildenafil and HIV-1 combination therapy. PMID- 10376646 TI - QT lengthening and arrhythmias associated with fexofenadine. PMID- 10376647 TI - CYP2A6 polymorphism, nicotine, and environmental nitrosamines. PMID- 10376648 TI - Betamethasone and placental vascular resistance. PMID- 10376649 TI - Jhum-jhum is a symptom. PMID- 10376650 TI - Propofol-infusion syndrome in children. PMID- 10376651 TI - Propofol-infusion syndrome in children. PMID- 10376652 TI - New formula to calculate mean aortic pressure? PMID- 10376653 TI - Duration of pregnancy and risk of breast cancer. PMID- 10376654 TI - Medical expert witnesses. PMID- 10376655 TI - Medical expert witnesses. PMID- 10376656 TI - Controversy over cancer chemotherapy in Japan. PMID- 10376659 TI - Kosovo's refugees. PMID- 10376660 TI - Debt relief and health charges in Mozambique. PMID- 10376661 TI - Weapons and the law. PMID- 10376663 TI - AIDS Web Gallery straddles worlds of art and medicine. PMID- 10376662 TI - Pooch protease paunch. PMID- 10376666 TI - The Nobel chronicles. 1958: George Wells Beadle (1903-89), Edward Lawrie Tatum (1909-75) and Joshua Lederberg (b 1925). PMID- 10376667 TI - Plant genomics. AB - The rapidity with which genomic sequences of the model plant Arabidopsis thaliana and soon of rice are becoming available has strongly boosted plant molecular biology research. Here, two main genomic fields will be discussed: the progress in different structural genome projects, such as mapping, sequencing, genome organization and comparative genomics, and the so-called functional genomics approaches to analyze the genome using such molecular tools as transcript profiling, micro-arrays, and insertional mutagenesis. In addition a section on bioinformatics is included. PMID- 10376668 TI - Learning from the genome sequence of Mycobacterium tuberculosis H37Rv. AB - Mycobacterium tuberculosis, the scourge of humanity, is one of the most successful and scientifically challenging pathogens of all time. To catalyse the conception of new prophylactic and therapeutic interventions against tuberculosis, and to enhance our understanding of the biology of the tubercle bacillus, the complete genome sequence of the most widely used strain, H37Rv, has been determined. Bioinformatic analysis led to the identification of approximately 4000 genes in the 4.41 Mb genome sequence and provided fresh insight into the biochemistry, physiology. genetics and immunology of this much feared bacterium. Genomic information is centralised in TubercuList (http://www.pasteur.fr/Bio/TubercuList/). PMID- 10376669 TI - Obligate intracellular parasites: Rickettsia prowazekii and Chlamydia trachomatis. AB - Transitions to obligate intracellular parasitism have occurred at numerous times in the evolutionary past. The genome sequences of two obligate intracellular parasites, Rickettsia prowazekii and Chlamydia trachomatis, were published last year. A comparative analysis of these two genomes has revealed examples of reductive convergent evolution, such as a massive loss of genes involved in biosynthetic functions. In addition, both genomes were found to encode transport systems for ATP and ADP, not otherwise found in bacteria. Here, we discuss adaptations to intracellular habitats by comparing the information obtained from the recently published genome sequences of R. prowazekii and C. trachomatis. PMID- 10376670 TI - Molecular and cellular activities of Helicobacter pylori pathogenic factors. AB - Stomach infection with pathogenic strains of Helicobacter pylori causes in some patients severe gastroduodenal diseases. These bacteria produce various virulence factors and, here, we review the recent acquisition on the biochemical mode of action of three major factors. We discuss the role of urease both as buffer of the stomach pH and as source of ammonia. The vacuolating toxin alters the endocytic pathway of non-polarized cells, inducing the release of acid hydrolases, the depression of extracellular ligand degradation and of antigen processing and, in the presence of ammonia, swelling of late-prelysosomal compartments. In polarized epithelial monolayers, vacuolating toxin induces an increase of the paracellular permeability, independent of vacuolation. The neutrophil activating protein induces the production of oxygen radicals in human neutrophils and could contribute to the damage of the stomach mucosa. The activities of these factors are discussed in terms of the need of the bacterium of increasing the supply of nutrients from the stomach lumen and from the mucosa. PMID- 10376671 TI - Extremophiles and their adaptation to hot environments. AB - Water-containing terrestrial, subterranean and submarine high temperature areas harbor a variety of hyperthermophilic bacteria and archaea which are able to grow optimally above 80 degrees C. Hyperthermophiles are adapted to hot environments by their physiological and nutritional requirements. As a consequence, cell components like proteins, nucleic acids and membranes have to be stable and even function best at temperatures around 100 degrees C. The chemolithoautotrophic archaeon Pyrolobus fumarii is able to grow at 113 degrees C and, therefore, represents the upper temperature border of life. For the first time, (vegetative) cultures of Pyrolobus and Pyrodictium are able to survive autoclaving. PMID- 10376672 TI - Folding and catalysis by the hairpin ribozyme. AB - The hairpin ribozyme undergoes a site-specific transesterification cleavage of the phosphodiester backbone. The natural form of the ribozyme is a four-way helical junction, where two arms contain unpaired loops. This folds by pairwise coaxial stacking of helical arms, and a rotation into an antiparallel conformation in which there is close association between the loops. This probably generates the local conformation required to facilitate the trajectory into an in line SN2 transition state. Folding is induced by the cooperative binding of at least two divalent metal ions, which are probably distributed between the junction and the loop-loop interface. The junction forms the structural scaffold on which the geometry of the ribozyme is built, and structural perturbation of the junction leads to impaired catalytic activity. PMID- 10376673 TI - Voltage-gated potassium channels: from hyperexcitability to excitement. AB - The superfamily of voltage-activated potassium channels may express structurally and functionally diverse voltage-activated potassium channels in the nervous system. The roles of some voltage-activated potassium channel types, e.g. rapidly inactivating (transiently active type) channels and muscarine sensitive muscarine sensitive channels, are beginning to be understood. They may significantly influence dendritic action-potential back-propagation, signal to noise ratios in presynaptic excitability or the responsiveness of a neuron to synaptic input. Inherited disorders related to changes in excitability (episodic ataxia, epilepsy, heart arrhythmia) or to defects in sensory perception (hearing loss) have been associated with mutations in a few voltage-activated potassium channel genes. Most likely, more voltage-activated potassium channel genes will be linked to related disorders in the near future. PMID- 10376674 TI - Modular PH and C2 domains in membrane attachment and other functions. AB - The pleckstrin homology and C2 domains are modular protein structures involved in mediating intermolecular interactions. Although they represent distinct domains, there are several parallels regarding their function and type of interactions in which they participate. Both domains are stable structural entities that incorporate variable regions which, in different proteins, can be adapted to perform a specific function through binding to membrane phospholipids or specific protein ligands. A number of recent examples illustrate the function of some of these domains in regulated membrane attachment, with an important role in many cellular signalling pathways. PMID- 10376675 TI - Structural information for explaining the molecular mechanism of protein biosynthesis. AB - Protein biosynthesis is controlled by a number of proteins external to the ribosome. Of these, extensive structural investigations have been performed on elongation factor-Tu and elongation factor-G. This now gives a rather complete structural picture of the functional cycle of elongation factor-Tu and especially of the elongation phase of protein biosynthesis. The discovery that three domains of elongation factor-G are structurally mimicking the amino-acylated tRNA in the ternary complex of elongation factor-Tu has been the basis of much discussion of the functional similarities and functional differences of elongation factor-Tu and elongation factor-G in their interactions with the ribosome. Elongation factor-G:GDP is now thought to leave the ribosome in a state ready for checking the codon-anticodon interaction of the aminoacyl-tRNA contained in the ternary complex of elongation factor-Tu. Elongation factor-G does this by mimicking the shape of the ternary complex. Other translation factors such as the initiation factor-2 and the release factor 1 or 2 are also thought to mimic tRNA. These observations raise questions concerning the possible evolution of G-proteins involved in protein biosynthesis. PMID- 10376676 TI - Transfer RNA modification: influence on translational frameshifting and metabolism. AB - Transfer RNA modification improves the rate of aa-tRNA selection at the A-site and the fitness in the P-site and thereby prevents frameshifting according to a new model how frameshifting occurs [Qian et al. (1998) Mol. Cell 1, 471-482]. Evidence that the presence of various modified nucleosides in tRNA also influences central metabolism, thiamine metabolism, and bacterial virulence is reviewed. PMID- 10376677 TI - Chloroplast precursor protein translocon. AB - Chloroplasts are believed to have originated from a photosynthetic, prokaryotic ancestor. As the result of endosymbiotic evolution, most of the genes of the endocytobiont were displaced to the host nucleus. Today's chloroplasts must import most of their proteins from the cytosol as precursors. Oligomeric protein complexes in the chloroplast outer and inner envelope membranes are responsible for the specific recognition and membrane translocation of precursor proteins. The translocon at the outer membrane of chloroplasts and the inner membrane of chloroplasts act jointly during the import process. Several translocon subunits have been partially characterized in their molecular structure and function. Initial evidence indicates the prokaryotic origin of some chloroplast translocon components. PMID- 10376678 TI - Mitochondrial assembly in yeast. AB - The yeast Saccharomyces cerevisiae is likely to be the first organism for which a complete inventory of mitochondrial proteins and their functions can be drawn up. A survey of the 340 or so proteins currently known to be localised in yeast mitochondria reveals the considerable investment required to maintain the organelle's own genetic system, which itself contributes seven key components of the electron transport chain. Translation and respiratory complex assembly are particularly expensive processes, together requiring around 150 of the proteins so far known. Recent developments in both areas are reviewed and approaches to the identification of novel mitochondrial proteins are discussed. PMID- 10376679 TI - Biogenesis of transport intermediates in the endocytic pathway. AB - Evidence is accumulating that membrane traffic between organelles can be achieved by different types of intermediates. Small (< 100 nm) and short-lived vesicles mediate transport from the plasma membrane or the trans-Golgi network to endosomes, and formation of these vesicles depends on specific adapter complexes. In contrast, transport from early to late endosomes is achieved by relatively large (approximately 0.5 microm), long-lived and multivesicular intermediates, and their biogenesis depends on endosomal COP-I proteins. Here, we review recent work on the formation of these different transport intermediates, and we discuss, in particular, coat proteins, sorting signals contained in cargo molecules and the emerging role of lipid in vesicle biogenesis. PMID- 10376680 TI - Endocytosis and intracellular transport of ricin: recent discoveries. AB - The plant toxin ricin has proven valuable as a membrane marker in studies of endocytosis as well as studies of different intracellular transport steps. The toxin, which consists of two polypeptide chains, binds by one chain (the B-chain) to both glycolipids and glycoproteins with terminal galactose at the cell surface. The other chain (the A-chain) enters the cytosol and inhibits protein synthesis enzymatically. After binding the toxin is endocytosed by different mechanisms, and it is transported via endosomes to the Golgi apparatus and the endoplasmic reticulum before translocation of the A-chain to the cytosol. The different transport steps have been analyzed by studying trafficking of ricin as well as modified ricin molecules. PMID- 10376681 TI - Editing of messenger RNA precursors and of tRNAs by adenosine to inosine conversion. AB - The double-stranded RNA-specific adenosine deaminases ADAR1 and ADAR2 convert adenosine (A) residues to inosine (I) in messenger RNA precursors (pre-mRNA). Their main physiological substrates are pre-mRNAs encoding subunits of ionotropic glutamate receptors or serotonin receptors in the brain. ADAR1 and ADAR2 have similar sequence features, including double-stranded RNA binding domains (dsRBDs) and a deaminase domain. The tRNA-specific adenosine deaminases Tad1p and Tad2p/Tad3p modify A 37 in tRNA-Ala1 of eukaryotes and the first nucleotide of the anticodon (A 34) of several bacterial and eukaryotic tRNAs, respectively. Tad1p is related to ADAR1 and ADAR2 throughout its sequence but lacks dsRBDs. Tad1p could be the ancestor of ADAR1 and ADAR2. The deaminase domains of ADAR1, ADAR2 and Tad1p are very similar and resemble the active site domains of cytosine/cytidine deaminases. PMID- 10376682 TI - Nuclear RNA export in yeast. AB - Eukaryotic cells massively exchange macromolecules (proteins and RNAs) between the nucleus and cytoplasm through the nuclear pore complexes. Whereas a mechanistic picture emerges of how proteins are imported into and exported from the nucleus, less is known about nuclear exit of the different classes of RNAs. However, the yeast Saccharomyces cerevisiae offers an experimental system to study nuclear RNA export in vivo and thus to genetically dissect the different RNA export machineries. In this review, we summarize our current knowledge and recent progress in identifying components involved in nuclear RNA export in yeast. PMID- 10376683 TI - Nuclear transport and transcriptional regulation. AB - Studies over the past 10 years have provided major insights into the molecular mechanisms responsible for active transport of macromolecules in and out of the nucleus. Nucleocytoplasmic transport pathways correspond to active and signal mediated processes that involve substrates, adaptors and receptors. Regulation of both nuclear import and nuclear export is mainly exerted at the level of transport complex formation and has emerged as one of the most efficient mechanisms to adapt gene expression to the cell environment by restricting the access of transcriptional regulators to their target genes. PMID- 10376684 TI - Initiation of DNA replication in eukaryotes: questioning the origin. AB - Although proteins involved in DNA replication in yeast have counterparts in multicellular organisms, the definition of an origin of DNA replication and its control in higher eukaryotes might obey to different rules. Origins of DNA replication that are site-specific have been found, supporting the notion that specific DNA regions are used to initiate DNA synthesis along metazoan chromosomes. However, the notion that specific sequences will define origins is still being debated. The variety and complexity of transcriptional programs that have to be regulated in multicellular organisms may impose a plasticity that would not be compatible with a fixed origin simply defined at the sequence level. Such a plasticity would be essential to developmental programs where the control of DNA replication could be more integrated to the control of gene expression than in unicellular eukaryotes. PMID- 10376685 TI - Protein kinases in control of the centrosome cycle. AB - The centrosome is the major microtubule nucleating center of the animal cell and forms the two poles of the mitotic spindle upon which chromosomes are segregated. During the cell division cycle, the centrosome undergoes a series of major structural and functional transitions that are essential for both interphase centrosome function and mitotic spindle formation. The localization of an increasing number of protein kinases to the centrosome has revealed the importance of protein phosphorylation in controlling many of these transitions. Here, we focus on two protein kinases, the polo-like kinase 1 and the NIMA related kinase 2, for which recent data indicate key roles during the centrosome cycle. PMID- 10376686 TI - Cytoskeleton cross-talk during cell motility. AB - Cell crawling entails the co-ordinated creation and turnover of substrate contact sites that interface with the actin cytoskeleton. The initiation and maturation of contact sites involves signalling via the Rho family of small G proteins, whereas their turnover is under the additional influence of the microtubule cytoskeleton. By exerting relaxing effects on substrate contact assemblies in a site- and dose-specific manner, microtubules can promote both protrusion at the front and retraction at the rear, and thereby control cell polarity. PMID- 10376687 TI - Lipid signaling in CD95-mediated apoptosis. AB - Ceramides play an important role mediating different cell responses such as proliferation, differentiation, growth arrest and apoptosis. They are released upon sphingomyelin hydrolysis which occurs after triggering of a number of cell surface receptors including CD95. Ceramide generation also regulates glycosphingolipid and ganglioside metabolism. In particular, ganglioside GD3 biosynthesis represents an important event for the progression of apoptotic signals generated by CD95 and mediated by ceramide in hematopoietic cells. PMID- 10376688 TI - Photodynamic therapy for nevus sebaceus with topical delta-aminolevulinic acid. PMID- 10376689 TI - Is pharmaceutical sponsorship of a dermatologic conference proper? PMID- 10376690 TI - Purple toes in a patient with end-stage rheumatoid arthritis. PMID- 10376691 TI - Equivalent therapeutic efficacy and safety of ivermectin and lindane in the treatment of human scabies. AB - OBJECTIVE: To compare the therapeutic efficacy and safety of ivermectin and lindane for the treatment of human scabies. DESIGN: Randomized, prospective, controlled, double-blind, "double-dummy," and parallel clinical study. SETTING: A single department of dermatology at a hospital in Buenos Aires, Argentina. PATIENTS: Patients were outpatients, hospitalized patients, and those referred to our hospital from nursing homes and asylums. Fifty-three patients had clinical signs and symptoms compatible with scabies. INTERVENTION: Patients received either a single oral dose of ivermectin (150-200 microg/kg of body weight) or a topical application of 1% lindane solution. Treatment was repeated after 15 days if clinical cure had not occurred. MAIN OUTCOME MEASURES: Clinical healing and adverse effects. RESULTS: Of 53 patients, 43 (81%) completed the study, 19 in the group treated with ivermectin and 24 in the group treated with lindane. At day 15, 14 patients (74%; 95% confidence interval, 48.8%-90.8%) in the group receiving ivermectin showed healing of their scabies and 13 patients (54%; 95% confidence interval, 32.8%-74.4%) in the group treated with lindane were healed. At 29 days, both treatments resulted in statistically equivalent therapeutic efficacy: 18 patients (95%; 95% confidence interval, 74.0%-99.9%) were healed with ivermectin and 23 patients (96%; 95% confidence interval, 78.9%, 99.9%) were healed with lindane (P<.02). Adverse effects from the treatments were few, mild, and transient. Results from laboratory tests showed no major abnormalities and no difference between treatments. CONCLUSIONS: Ivermectin is as effective as lindane for the treatment of scabies. Ivermectin is simpler to use and, therefore, is a promising tool to improve compliance and to control infestations. PMID- 10376692 TI - Hair density in African Americans. AB - BACKGROUND: The meager data on normal hair density in humans have been gathered from a predominantly white population. Examination of scalp biopsy specimens from African Americans suggests that hair density in this group may be lower than in whites. This study was performed to quantify any differences between white and African American patients. DESIGN: A retrospective case series of subjects who had undergone a biopsy of clinically healthy scalp skin. The 4-mm punch biopsy specimens were sectioned, and all follicles contained within the specimens were counted at various levels (suprabulbar, isthmus, and infundibulum) to arrive at the number and type of hairs present. SETTING: Outpatient clinic in a tertiary care medical center. PATIENTS: A consecutive sample of 22 African American and 12 white patients with clinically healthy scalp skin specimens that were studied and compared with previously reported data. MAIN OUTCOME MEASURES: Patients' age and total number of follicles, terminal follicles, vellus follicles, terminal anagen hairs, and terminal telogen hairs. RESULTS: Total hair density (number of follicles per 4-mm punch biopsy specimen) and total number of terminal follicles and terminal anagen hairs were significantly lower in African Americans (P<.001) than in whites and in a previously reported, predominantly white, population. CONCLUSIONS: Hair density in African Americans is significantly lower than that in whites, which must be taken into consideration when evaluating a biopsy specimen from an African American patient. Data previously collected from white patients may not provide adequate guidance when evaluating scalp biopsy specimens from African Americans and could lead to an incorrect diagnosis. PMID- 10376693 TI - Effect of treatment of Helicobacter pylori infection on rosacea. AB - OBJECTIVE: To evaluate the clearing and intensity of symptoms of rosacea 60 days after the treatment of Helicobacter pylori infection. DESIGN: Randomized, double blind, placebo-controlled clinical trial. SETTING: The dermatology section of a large multispecialty clinic in the North Central United States. PARTICIPANTS: Men and women older than 25 years with active signs of rosacea who tested positive for H pylori with both the rapid whole blood test and the urea breath test. INTERVENTION: Treatment of H pylori infection with 14-day therapy using clarithromycin. 500 mg orally 3 times a day, and omeprazole, 40 mg orally once a day. MAIN OUTCOME VARIABLES: The extent and intensity of rosacea as measured by the number of papules and pustules and the extent and intensity of erythema and telangiectasia. RESULTS: Three hundred twenty patients presented with rosacea. For 50 patients, the results of a urea breath test were positive for H pylori, and 44 patients were enrolled in the study. Rosacea abated in almost all patients, but none were cured. Notably, lessening of rosacea for patients treated for H pylori was not significantly better than for the control cohort. CONCLUSIONS: Rosacea abated in most participants in this study, whether they were in the treatment or the control cohort. There was no statistical difference when the results of active treatment were compared with those of placebo. Treating H pylori infection has no short-term beneficial effect on the symptoms of rosacea to support the suggested causal association between H pylori infection and rosacea. PMID- 10376694 TI - High-intensity flashlamp photoepilation: a clinical, histological, and mechanistic study in human skin. AB - OBJECTIVE: To examine the clinical, histological, and immunohistological effects of flashlamp photoepilation. DESIGN: Nonrandomized control trial with blinded histological study and follow-up of 1 to 20 months. SETTING: Private academic practice. SUBJECTS: Sixty-seven subjects (10 males and 57 females) with areas of excess body hair. INTERVENTIONS: Single (9 subjects) or multiple (58 subjects) treatments (noncoherent, 590-1200 nm, 2.9-3.0 milliseconds, 40-42 J/cm2) to hairy skin. From subjects given a single treatment, biopsy samples were taken immediately after treatment and at different intervals for up to 20 months. MEAN OUTCOME MEASURES: Clinical measures include hair counts and morphologic features before and after treatment. Histological measures include terminal-vellus and anagen-other ratios, hair shaft diameter, and morphologic features (routine and immunohistochemical detection of bcl-2, bax, p53, Ki67, cyclin D1, and hsp70) before and after treatment. RESULTS: Mean hair loss after photoepilation was 49%, 57%, and 54% for a single treatment and 47%, 56%, and 64% for multiple treatments at follow-up of less than 3 months, 3 to less than 6 months, and 6 months or longer, respectively (P<.05 for all comparisons). Transient erythema was seen in all subjects; no scarring occurred. Histologically, treatment caused morphologic damage confined to hair follicles and shafts. Terminal-vellus and anagen-telogen ratios, mean hair shaft diameter, and immunohistochemical profiles were not significantly modified by treatment. Treatment did not alter other skin adnexa, epidermis, or vessels. CONCLUSIONS: Flashlamp treatment leads to significant, longlasting epilation. The predominant mechanism seems to be via selective photothermal damage to large, pigmented hair follicles rather than induction of a programmed state of follicular cycle arrest or follicular miniaturization. PMID- 10376695 TI - Treatment of port-wine stains with a noncoherent pulsed light source: a retrospective study. AB - OBJECTIVE: We investigated whether a noncoherent intense pulsed light source (IPLS) would be effective in therapy of port-wine stains (PWSs). DESIGN: To evaluate the efficacy in treatment of PWSs with IPLS, a retrospective study was initiated. SETTING: The data were collected by physicians working in private practices and departments of university hospitals and medical centers, respectively. PATIENTS: A total of 37 randomly selected patients with a total of 40 PWSs were included in the study. Clinical PWS characteristics recorded were color and location of the PWS. INTERVENTIONS: All patients were treated with IPLS. MAIN OUTCOME MEASURES: Data collected included treatment parameter (filters, pulse duration, fluence, and pulse sequencing), percentage of clearance, and side effects (purpura, blisters, crusting, altered pigmentation, and scarring). RESULTS: Good and complete (70%-100%) clearance was achieved in 28 of 40 PWSs treated with IPLS. The average number of treatment sessions in PWSs reaching 100% clearance included 4.0 for pink PWSs and 1.5 for red PWSs. The average number of sessions for purple PWSs reaching good clearance (70%-99%) was 4.2 sessions. Parameters used most frequently were 515- and 550-nm cut-off filters, pulse duration of 2.5 to 5.0 milliseconds, and fluences of 24 to 60 J/cm2. Side effects included purpura in 133 (76%), superficial blisters in 14 (8%), and crusting in 35 (20%). Transient pigmentation changes were seen in 10.8% of patients (hypopigmentation in 3 [8.1%], hyperpigmentation in 1 [2.7%]). No scarring was observed. CONCLUSION: Intense pulsed light source presents an effective and safe method for treating PWSs, especially purple PWSs. PMID- 10376696 TI - Pinta in Austria (or Cuba?): import of an extinct disease? AB - BACKGROUND: Pinta, 1 of the 3 nonvenereal treponematoses, is supposed to be extinct in most areas in South and Central America, where it was once endemic. Only scattered foci may still remain in remote areas in the Brazilian rain forest, and the last case from Cuba was reported in 1975. OBSERVATION: A native Austrian woman, who had lived for 7 years in Cuba and was married to a Cuban native, developed a singular psoriasiform plaque on her trunk and several brownish papulosquamous lesions on her palms and soles during a visit to her home in Austria. Positive serological findings for active syphilis and the detection of spirochetes in the trunk lesion indicated early secondary syphilis, but an extensive case history and the clinical appearance fulfilled all criteria for pinta. CONCLUSION: The acquisition of a distinct clinical entity, pinta, in a country where it was formerly endemic but now is believed to be extinct raises the question of whether the disease is in fact extinct or whether syphilis and pinta are so similar that no definite distinction is possible in certain cases. PMID- 10376697 TI - Cryogen spray cooling in combination with nonablative laser treatment of facial rhytides. AB - BACKGROUND: Cryogen spray cooling can be used to provide epidermal protection while still achieving spatially selective photocoagulation in the upper dermis. The objective of this study is to determine the efficacy and safety of cryogen spray cooling in combination with a nonablative Nd:YAG (lambda = 1320 nm) laser treatment of facial rhytides in human volunteers. OBSERVATIONS: Thirty-five adults with bilateral periorbital rhytides were treated with cryogen spray cooling in combination with 3 nonablative laser treatments performed sequentially at intervals of 2 weeks. Small but statistically significant improvements were noted in the mild, moderate, and severe rhytid groups 12 weeks after the final laser treatment. A final assessment performed 24 weeks after the last treatment showed statistically significant improvement only in the severe rhytid group. The procedure was found to be safe; 4 sites (5.6%) developed transient hyperpigmentation. Two sites (2.8%) subsequently developed barely perceptible pinpoint pitted scars. CONCLUSIONS: Cryogen spray cooling is a safe and effective method for protecting the epidermis during nonablative laser treatment of facial rhytides thereby avoiding much of the morbidity associated with other resurfacing procedures. Minor improvements in rhytides can be achieved with the current technology. Optimization of treatment parameters may further improve these results. PMID- 10376698 TI - Iatrogenic cutaneous injuries in the neonate. AB - BACKGROUND: Iatrogenic cutaneous injuries of the neonate have decreased in number in the last 30 years because of changes in the medical procedures during the prenatal, perinatal, and postnatal periods. OBSERVATIONS: The emergence of such cutaneous injuries derived from the use of instruments, blunt and sharp, from manual manipulations, from medications, and from hesitation or abstaining from intervention. CONCLUSION: The dermatologist, unfamiliar with handling of neonates in the nursery, should be capable of recognizing and dealing with these phenomena when encountered in the acute stage or with their residue. PMID- 10376700 TI - A slowly growing tender plaque of the palm. PMID- 10376699 TI - Ivermectin: a new therapeutic weapon in dermatology? PMID- 10376701 TI - Red eroded nodule on the foot. PMID- 10376702 TI - Congenital painful pedal mass. PMID- 10376703 TI - Asymptomatic scalp nodule present for 20 years. PMID- 10376704 TI - Lichen nitidus actinicus. PMID- 10376705 TI - Prognostic factors in leukocytoclastic vasculitis: what is the role of antineutrophil cytoplasmic antibody? PMID- 10376706 TI - Persistent periungual erythema with telangiectasia: red fingers syndrome. PMID- 10376707 TI - Antibiotic prophylaxis. PMID- 10376709 TI - The "hairless" gene in mouse and man. PMID- 10376708 TI - Cost-effectiveness of methotrexate and Goeckerman therapy: a flawed analysis. PMID- 10376710 TI - Fluorescence-activated cell sorter analysis in patients with cutaneous lupus erythematosus. PMID- 10376711 TI - STIR magnetic resonance imaging: a noninvasive method for detection and follow-up of dermatomyositis. PMID- 10376712 TI - Antibiotic sensitivity of Propionibacterium acnes isolates studied in a skin clinic in Singapore. PMID- 10376713 TI - Treatment of pemphigus vulgaris and bullous pemphigoid with mycophenolate mofetil monotherapy. PMID- 10376714 TI - Delayed appearance of livedo reticularis in 3 cases with a cholesterol embolism. PMID- 10376715 TI - Sodium benzoate-induced acute leukocytoclastic vasculitis with unusual clinical appearance. PMID- 10376717 TI - Dermatosis neglecta: dirt crusts simulating verrucous nevi. PMID- 10376716 TI - A novel human papillomavirus DNA sequence related to epidermodysplasia verruciformis-associated types isolated from recurrent scar carcinoma. PMID- 10376718 TI - Failure of partial cementation to achieve implant stability and bone ingrowth: a long-term roentgen stereophotogrammetric study of tibial components. AB - Thirty patients with gonarthrosis were operated on with the PCA primary total knee prosthesis and had the tibial component fixed to the bone by partial cementation. In the first two groups of patients, cementation was by a peripheral rim of high and low-viscosity cement, respectively. In the third group, the pegs were cemented with the low-viscosity cement. Follow-up was performed with use of clinical parameters and roentgen stereophotogrammetric analysis. Clinically, the series was successful apart from a problem with tibial component wear, necessitating revision in five patients. At 8 years, the mean Hospital for Special Surgery score was 81 points. Venn-diagram scores revealed four failures and three acceptable cases; the remaining cases were satisfactory. Apart from one loose patellar component, there was no mechanical loosening. Roentgen stereophotogrammetric analysis showed that the tibial components moved relative to the bone; this indicated fibrous tissue fixation, which was corroborated histologically in two patients. The objective, to achieve bone ingrowth, was thus not successfully met. Radiolucent lines were consistently seen, and their size correlated with the migration as measured by roentgen stereophotogrammetric analysis. Furthermore, five continuously migrating prostheses showed significantly larger radiolucent zones than the prostheses that migrated only initially, and they were less well bonded to the bone at 1 year. In conclusion, partial cementation does not appear to be a way to achieve bone ingrowth in porous-coated implants under load. PMID- 10376719 TI - Mixed-mode fracture toughness of the cobalt-chromium alloy/polymethylmethacrylate cement interface. AB - Mechanical debonding of the stem/cement interface has been implicated in the failure process of cemented femoral hip components. The nature of this failure process remains poorly understood due, in part, to limited understanding of how interfacial debonding occurs in response to a wide range of loading conditions. The purpose of this investigation was to determine the fracture toughness of the cobalt-chromium alloy/polymethylmethacrylate interface under mixed-mode loading conditions. The hypothesis was that the critical energy release rate was dependent on the phase angle of the crack tip and that the fracture response would be significantly different for a smooth compared with rough interface surface. A novel in-plane shear test fixture was developed with use of a combination of finite element and experimental fracture-mechanics tests. A wide range (-65-60 degrees) of phase angles was determined with the in-plane shear test and a clamped cantilever-beam test. Sixty experimental tests were performed for cobalt-chromium alloy bars with a plasma-sprayed coating or a precoat of polymethylmethacrylate over a satin-finished surface. For the specimens with the plasma-sprayed coating, critical energy release rates (500-700 J/m2) were not a function of the phase angle of the crack tip. In contrast, critical energy release rates (15-80 J/m2) were found to be strongly affected by the phase angle for the specimens precoated with polymethylmethacrylate. The critical energy release rate for specimens with the plasma-sprayed surface was significantly (p < 0.01) greater than for those precoated with polymethylmethacrylate. The critical energy release rate increased markedly with the phase angle of the crack tip for the specimens precoated with polymethylmethacrylate. The results suggest that the failure response of a stem with a plasma-sprayed surface may be insensitive to the loading angle of the crack tip, whereas a stem precoated with polymethylmethacrylate may be more likely to debond under tensile opening loading. PMID- 10376720 TI - Effect of molecular weight, calcium stearate, and sterilization methods on the wear of ultra high molecular weight polyethylene acetabular cups in a hip joint simulator. AB - Orthopaedic surgeons must currently choose from several types of ultra high molecular weight polyethylene acetabular cups that differ in their material properties and in the methods used for their sterilization. Information on the wear resistance of these different cups may help in the selection process. This study included two separate tests for wear run on a hip simulator to investigate the effect of molecular weight, calcium stearate, and sterilization methods on the wear resistance of ultra high molecular weight polyethylene acetabular cups. Test 1 revealed nearly identical wear rates for acetabular cups with molecular weights in two distinct ranges, as well as for cups with molecular weights in the same range but with or without calcium stearate added. In Test 2, cups that were sterilized in air with gamma irradiation exhibited lower rates of wear than those sterilized with ethylene oxide, presumably due to the crosslinking induced by the irradiation. In addition, cups that were irradiated while packed in a partial vacuum to minimize oxygen absorbed in the surface layer initially showed lower rates of wear than those irradiated in air, with the wear rates becoming similar as wear penetrated the more oxidized surface layer and the more crosslinked subsurface region. Because these tests were run a few months after the irradiation, the potential effects of long-term oxidation of any residual free radicals in the irradiated materials could not be taken into account. After artificial aging to accelerate oxidative degradation of the materials, the wear rates could be markedly different. Analyses performed after wear indicated that the irradiated (i.e., crosslinked) cups exhibited a smaller proportion of, as well as shorter, fibrils in the wear debris and an increased crystallinity and melting temperature and that gamma irradiation in the low-oxygen environment reduced the level of oxidation and increased the level of crosslinking in the surface region of the cups. PMID- 10376721 TI - Effect of serum proteins on osteoblast adhesion to surface-modified bioactive glass and hydroxyapatite. AB - Previous studies indicate that modification of the surface of porous bioactive glass promotes osteoblast function. We hypothesize that bone formation on treated bioactive glass is due to the selective adsorption of serum attachment proteins. To test this hypothesis, we examined the profile of proteins adsorbed to treated bioactive glass and compared these proteins with those adsorbed to untreated bioactive glass and porous hydroxyapatite. Porous bioactive glass was treated with Tris-buffered electrolyte solution to generate a calcium phosphate-rich surface layer and then immersed in tissue-culture medium containing 10% serum. Proteins adsorbed to the ceramic surfaces were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot analysis. Porous hydroxyapatite bound a higher amount of total protein than did the other substrates. However, surface-modified porous bioactive glass adsorbed more fibronectin than did hydroxyapatite. The effect of serum-protein adsorption on osteoblast adhesion to bioactive glass and hydroxyapatite was also evaluated. Cell adhesion to porous bioactive glass that was surface-modified and serum treated was significantly greater than to porous bioactive glass that was either surface-modified or serum-treated. Furthermore, cell adhesion to porous bioactive glass treated to form the dual layer of calcium phosphate and serum protein was significantly higher than adhesion to porous hydroxyapatite with adsorbed serum protein. Results of the study strongly suggest that adsorption of serum fibronectin to the surface of modified porous bioactive glass coated with calcium phosphate may be responsible for enhanced osteoblast adhesion. PMID- 10376722 TI - Mechanical behavior of human trabecular bone after overloading. AB - With the etiology of osteoporotic fractures as motivation, the goal of this study was to characterize the mechanical behavior of human trabecular bone after overloading. Specifically, we quantified the reductions in modulus and strength and the development of residual deformations and determined the dependence of these parameters on the applied strain and apparent density. Forty cylindrical specimens of human L1 vertebral trabecular bone were destructively loaded in compression at 0.5% strain per second to strains of up to 3.0% and then immediately unloaded to zero stress and reloaded. (An ancillary experiment on more readily available bovine bone had been performed previously to develop this testing protocol.) In general, the reloading stress-strain curve had a short initial nonlinear region with a tangent modulus similar to Young's modulus. This was followed by an approximately linear region spanning to 0.7% strain, with a reduced residual modulus. The reloading curve always approached the extrapolated envelope of the original loading curve. Percent modulus reduction (between Young's and residual), a quantitative measure of mechanical damage, ranged from 5.2 to 91.0% across the specimens. It increased with increasing plastic strain (r2 = 0.97) but was not related to modulus or apparent density. Percent strength reduction, in the range of 3.6-63.8%, increased with increasing plastic strain (r2 = 0.61) and decreasing apparent density (r2 = 0.23). The residual strains of up to 1.05% depended strongly on applied strain (r2 = 0.96). Statistical comparisons with previous data for bovine tibial bone lend substantial generality to these trends and provide an envelope of expected behavior for other sites. In addition to providing a basis for biomechanical analysis of the effects of damage in trabecular bone at the organ level, these findings support the concept that occasional overloads may increase the risk of fracture by substantially degrading the mechanical properties of the underlying trabecular bone. PMID- 10376724 TI - Effect of lengthening rate on angiogenesis during distraction osteogenesis. AB - This study investigated the angiogenic response to four varying rates (0.3, 0.7, 1.3, and 2.7 mm/day) of distraction in a rabbit model of leg-lengthening. Immunostaining was performed with use of specific antibodies to type-IV collagen and endothelial cell antigen to examine semiquantitatively the presence of blood vessels in the developing tissues. With use of the Chalkley counting method, the greatest number of positive-staining blood vessel cells was found in the central fibrous zone of the groups that underwent lengthening at 0.7 and 1.3 mm/day compared with any other zone in any group (p < 0.05, t test). There were no statistical differences in the positive labeling indices in the mineralization front and the new bone zone adjacent to the mineralization front in any of the groups. However, the decrease in the number of positive-staining blood vessel cells in the new bone zone distant to the mineralization front compared with any other zone in any group was statistically significant. The results suggest that during distraction osteogenesis, the precursor cells of new capillaries were present in abundance within the fibrous interzone. These cells connected into the capillary network at the junction of the mineralization front and the fibrous zone. The angiogenic response was weaker in the more mature regions within the new bone zones. A slow rate of distraction (0.3 mm/day) did not maximally stimulate angiogenesis in the central fibrous zone, whereas high rates (2.7 mm/day) appeared to impair this response. In this model of distraction osteogenesis, the vascularization process in the central fibrous zone was maximally stimulated at distraction rates of 0.7 and 1.3 mm/day. PMID- 10376723 TI - Increased expression of matrix metalloproteinase-1 in osteocytes precedes bone resorption as stimulated by disuse: evidence for autoregulation of the cell's mechanical environment? AB - An in vivo animal model of bone adaptation was used to examine a possible role for matrix metalloproteinase-1 in the local mediation of bone remodeling: to corrode the coupling of osteocytes to the matrix in an attempt to autoregulate the cell's perception of its mechanical environment. Twelve young (12-16 months old) skeletally mature turkeys were separated into groups to be studied for stimulus periods of either 3 or 30 days. In each animal, the left ulna was functionally isolated and subjected to either disuse or 3,000 microstrain at 1 Hz for 10 minutes per day. The right ulna remained intact and served as an intra animal control. No significant differences in bone area were detected at 3 days; however, ulnae subjected to disuse lost 8 +/- 4% (+/-SD) of bone area by 30 days. Over the same period, ulnae subjected to the mechanical stimulus gained 21 +/- 9% of bone area. With use of in situ reverse transcription-polymerase chain reaction, less than 2% of the osteocytes examined from the intact control ulnae stained positively for matrix metalloproteinase-1 mRNA. An antibody raised against matrix metalloproteinase-1 revealed no positively labeled osteocytes in the intact ulnae. This low percentage of osteocytes expressing matrix metalloproteinase-1 mRNA was similar to that seen in ulnae subjected to the osteogenic mechanical stimuli. In contrast, ulnae subjected to either 3 or 30 days of disuse showed evidence of matrix metalloproteinase-1 mRNA activity in a high percentage of osteocytes (89 +/- 5 and 66 +/- 8%, respectively; each time point significantly different from intact ulnae, as well as from each other, p < 0.05). The percentage of osteocytes labeled with the anti-matrix metalloproteinase-1 antibody was also highly elevated following 3 days of disuse (74 +/- 17%). These data demonstrate that an early response of bone to disuse is the upregulation of matrix metalloproteinase-1 activity in osteocytes. It is proposed that this upregulation of collagenase activity is indicative of the cell's degradation of coupling to the matrix, and it thus reflects the osteocyte's regulation of its own mechanical environment. We believe that such autoregulation of the osteocyte's physical environment will accommodate subtle changes in the bone's functional environment without the need to add or resorb bone tissue. PMID- 10376725 TI - Assessment of subchondral bone blood flow in the rabbit femoral condyle using the laser speckle method. AB - The laser speckle method is a new form of flowmetry that can obtain a two dimensional distribution of blood flow in tissue. This method is a noncontact, simple, and rapid technique that may aid in the diagnosis of osteonecrosis. We investigated whether the subchondral bone blood flow within the femoral condyles of rabbits could be measured by the laser speckle method. The hydrogen washout method was chosen as a comparison technique because of its ability to allow repetitive measurements of blood flow in various conditions in one rabbit and because of its reliability, which already has been established. We simultaneously measured the bone blood flow in 20 femoral condyles of 10 rabbits with the laser speckle and hydrogen washout methods and found a significant correlation between the blood flow levels with use of these two methods. For the clinical application of the laser speckle method, we also investigated the influence of cartilage thickness on the measurements and the depth in the bone to which blood flow could be measured with this method. A cartilage thickness of 0.2 mm did not influence the measurement of the bone blood flow, and the depth in the bone to which the laser speckle method could be used was approximately 2 mm. PMID- 10376726 TI - Experimental acute hematogenous osteomyelitis in mice. I. Histopathological and immunological findings. AB - This study investigated immunological responses to Staphylococcus aureus bone infection. Because considerable immunological information is available on the mouse, a murine model of acute hematogenous osteomyelitis was established. Osteomyelitis was created in the proximal tibia of C3H/HeJ mice by a tibial epiphyseal fracture followed by intravenous bacterial inoculation with Staphylococcus aureus (strain LS-1). Swelling and warmth of the limb was found, and following limb exposure, abscess formation was evident in the proximal tibia. Histological examination revealed distortion primarily at the hypertrophic zone of the physis and polymorphonuclear leukocyte infiltration throughout the damaged area of the proximal tibia. Local infection was demonstrated at the fracture site, evidenced by the recovery of Staphylococcus aureus following microbiological analysis of tissue specimens. Polymerase chain reaction was utilized to detect 16S ribosomal prokaryotic nucleic acid to demonstrate that the diagnosis of osteomyelitis could be established in the absence of conventional microbiological techniques. The infected mice had an increase of circulating large leukocytes (granulocytes) and an elevation of total serum immunoglobulin. Flow cytometry revealed significant increases in splenic B lymphocytes and in lymph-node CD4+ T lymphocytes. These results indicate that an experimental model of acute hematogenous osteomyelitis that closely resembles the pathology of the disease in humans may be consistently induced in mice. Furthermore, marked immunological changes may be observed in response to the Staphylococcus aureus bone infection. PMID- 10376728 TI - Increased interleukin-1beta production in the synovium of glenohumeral joints with anterior instability. AB - Macroscopic synovitis of the glenohumeral joint is frequently seen during arthroscopy in patients with anterior instability. Interleukin-1beta is known to be expressed in inflamed tissue, to correlate with the magnitude of inflammation, and to affect articular cartilage in the joint. We hypothesized that chronic synovitis may occur in the glenohumeral joint in patients with anterior instability. The purpose of this study was to examine the expression of interleukin-1beta in the synovium of the glenohumeral joint with anterior instability and to discuss its clinicopathologic significance. Specimens of synovial tissue around the greater tuberosity in the subacromial synovium (as controls) and around the rotator interval in the glenohumeral synovium were obtained from 10 patients who had anterior instability without signs of subacromial impingement. Semiquantitative reverse transcriptase-polymerase chain reaction was used to compare the levels of interleukin-1beta mRNA expression in the glenohumeral joint with those in the subacromial bursa. We also employed immunohistochemistry and in situ reverse transcriptase-polymerase chain reaction to detect the cells producing interleukin-1beta protein and mRNA. The levels of interleukin-1beta mRNA expression were significantly higher in the glenohumeral joint than in the subacromial bursa (p < 0.01). Histology showed nonspecific inflammation in all 10 samples of glenohumeral synovium, whereas no inflammation was seen in seven of 10 samples of subacromial synovium. Immunohistochemistry identified interleukin-1beta protein in the vessels and inflammatory and synovial cells (from lining to sublining layers) in synovium of the glenohumeral joint, whereas immunoreactivity was negative in seven subacromial bursa. The remaining three synovial specimens of subacromial bursa, however, showed positive immunoreactivity that was unremarkable and confined around the vessels. In situ reverse transcriptase-polymerase chain reaction was exclusively performed in the synovial specimens of the glenohumeral joint, which exhibited a positive reaction (in the same kinds of cells as seen with immunohistochemistry) in the lining and sublining layers and to a lesser extent in the stroma. Thus, our data confirmed the increased production of interleukin-1beta in the synovium of the glenohumeral joint in patients with anterior instability, suggesting the presence of chronic inflammation at the site. We conclude that this chronic synovitis may be partly associated with the development of dislocation arthropathy in the long term. PMID- 10376730 TI - Estrogen level alters the failure load of the rabbit anterior cruciate ligament. AB - We hypothesized that increasing the level of circulating serum estrogen would decrease the load at failure for the rabbit anterior cruciate ligament. We developed an animal model in which hormonal manipulations could be correlated with load at failure for the anterior cruciate ligament. Sixteen New Zealand White ovariectomized rabbits were matched and divided into two groups. The eight rabbits that were not treated with the estrogen supplement (Group O) and the eight that were treated with the supplement (Group E) were housed for 30 days. Serum estrogen levels were measured. The knees were stripped of all soft tissue, and the load at failure for the anterior cruciate ligament was measured in a materials testing machine with a displacement rate of 0.5 mm/sec. The load at failure for all 16 specimens in Group E (446 +/- 54 N) (mean +/- SD) was significantly reduced (p = 0.02) compared with that for the nine specimens in Group O (503 +/- 48 N). It is recognized that an increased number of anterior cruciate ligament injuries occurs in female athletes. Although the mechanism responsible for failure of the anterior cruciate ligament in women is yet to be defined, this experiment suggests that estrogen may alter ligament strength. PMID- 10376727 TI - Experimental acute hematogenous osteomyelitis in mice. II. Influence of Staphylococcus aureus infection on T-cell immunity. AB - A murine model of acute hematogenous osteomyelitis was used to study the immune response following Staphylococcus aureus infection and to examine the hypothesis that the bacteria may modify T-cell responses due to the production of bacterial enterotoxins with mitogenic or superantigenic activity. Lymph-node T cell receptor expression was assessed with use of flow cytometry and reverse transcription-polymerase chain reaction techniques, and increased apoptosis (programmed cell death) in T-cell subsets was monitored. The expression and levels of circulating cytokines and T-cell cytokines within tissues surrounding the damaged area of the proximal tibia were also investigated. Analysis of T-cell receptors in experimental osteomyelitis revealed two distinct patterns of T-cell evolution during the disease. Certain T-cell subsets (Vbeta2, Vbeta3, Vbeta9, and Vbeta10) were activated and expanded during the first 24 hours after infection; they reached maximum levels 6 days after infection, followed by a return to pre infection levels. In contrast, other T-cell subsets (Vbeta11, Vbeta12, Vbeta13, Vbeta14, and Vbeta16) contracted during the first 24 hours after infection, followed by expansion to a maximum level 9 days after infection. Activation and proliferation of T-cell subsets (notably Vbeta14 T cells) was followed by apoptosis, suggesting that staphylococcal bone infection caused superantigenic like effects on the mouse immune system. Analysis of cytokine responses in local tissue revealed that the T-cell cytokines interleukin-2 and interferon-gamma showed a late and relatively short activation pattern compared with the inflammatory cytokines interleukin-1, interleukin-6, and tumor necrosis factor alpha. The results suggest that Staphylococcus aureus bone infection may undermine the antibacterial immune response through downregulation of T-cell immunity and immune-cytokine production, which could increase the severity of the systemic infection and local osseous destruction that occur with acute hematogenous osteomyelitis. PMID- 10376729 TI - Characterization of chemotactic migration and growth kinetics of canine knee ligament fibroblasts. AB - Migration and proliferation of ligament fibroblasts are essential for the healing of ligament injuries. This study was designed to evaluate the migration of intraarticular (anterior and posterior cruciate) and extraarticular (medial and lateral collateral) ligament fibroblasts in response to cytokines and to determine the effect of cell passage on cell proliferation. Recombinant human platelet-derived growth factor, hepatocyte growth factor/scatter factor, and bone morphogenic protein-2 stimulated the migration of all ligament cells in a dose dependent manner, with optimal migration at 10 ng/ml. Recombinant human epithelial growth factor preferentially stimulated the migration of intraarticular ligament fibroblasts, whereas recombinant human interleukin-1 was more effective with extraarticular ligament fibroblasts. Recombinant human insulin-like growth factor-1, insulin-like growth factor-2, transforming growth factor-beta, and fibroblast growth factor had no significant effect on the migration of ligament-derived fibroblasts. These data suggest that specific cytokines stimulate the migration of knee ligament fibroblasts and provide a rationale for possible therapeutic approaches to optimize ligament healing. Fibroblasts derived from the anterior cruciate ligament have been shown to proliferate at a slower rate than those derived from the medial collateral ligament. We have extended these observations and have demonstrated that fibroblasts from both the posterior and anterior cruciate ligaments proliferate at a slower rate than lateral and medial collateral ligament-derived fibroblasts. The differences between the growth rates of intraarticular and extraarticular fibroblasts become insignificant with serial passaging of the cells. PMID- 10376731 TI - Stiffness and muscle function with age and reduced muscle use. AB - Changes in passive muscle stiffness with age and disuse were assessed in male Fischer-344 and Brown Norway rats. Three groups of rats were studied: young (approximately 7 months old), old (approximately 33 months old), and old that had undergone 2 weeks of hindlimb unweighting, a model of reduced muscle use. Four hindlimb muscles were examined: the soleus (postural), plantaris (locomotor), extensor digitorum longus (nonpostural), and peroneus longus (nonpostural). Supramaximal stimuli elicited peak tetanic tensions throughout the available range of motion (amount of muscle elongation before the maximal attainable contractile or tetanic tension is obtained) for each muscle, permitting the creation of length-tension curves. Passive tension (amount encountered at each millimeter of change in muscle length) was also recorded throughout the available range of motion and was unchanged with aging and unchanged or reduced with hindlimb unweighting; muscle stiffness remained unchanged under both conditions. Passive tension, however, accounted for a greater proportion of total tension with age and particularly with hindlimb unweighting. A significant loss in muscle range of motion, resulting in a leftward shift in the length-tension curve, occurred with aging in only the plantaris. Hindlimb unweighting resulted in a marked loss in muscle range for all four muscles studied, suggesting that the remaining muscle force was constrained to a very small arc. Significant declines in muscle mass and peak contractile tension, associated with age and hindlimb unweighting, were observed for all four muscles. PMID- 10376732 TI - Influences of inflation rate and duration on vasodilatory effect by intermittent pneumatic compression in distant skeletal muscle. AB - Previous study has demonstrated that application of intermittent pneumatic compression on legs can cause vasodilation in distant skeletal muscle at the microcirculation level. This study evaluated the influence of inflation rate and peak-pressure duration on the vasodilatory effects of intermittent pneumatic compression. The cremaster muscles of 50 male rats were exposed and divided into five groups of 10 each. A specially designed intermittent pneumatic-compression device was applied in a medial-lateral fashion to both legs of all rats for 60 minutes, with an inflation rate and peak-pressure duration of 0.5 and 5 seconds, respectively, in group A, 5 and 0 seconds in group B, 5 and 5 seconds in group C, 10 and 0 seconds in group D, and 10 and 5 seconds in group E. Diameters of arterial segments were measured in vessels of three size categories (10-20, 21 40, and 41-70 microm) for 120 minutes. The results showed that the greatest increase in diameter was produced by intermittent pneumatic compression with the shortest inflation rate (0.5 seconds). A moderate increase resulted from compression with an inflation rate of 5 seconds, and no effective vasodilation occurred during compression with the longest inflation rate (10 seconds). When the groups with different inflation rates but the same peak-pressure duration were compared, there was a significant difference between any two groups among groups A, C, and E and between groups B and D. When the groups with different peak-pressure durations but the same inflation rate were compared, compression with a peak-pressure duration of 5 seconds caused a generally similar degree of diameter change as did compression without inflation at peak pressure. The findings suggest that inflation rate plays an important role in the modulation of distant microcirculation induced by intermittent pneumatic compression whereas peak-pressure duration does not significantly influence the vasodilatory effects of the compression. This may be due to the fact that rapid inflation produces a significant increase in shear stress on the vascular wall, which stimulates vascular endothelium to release nitric oxide, causing systemic vasodilation. PMID- 10376733 TI - Mechanically induced calcium waves in articular chondrocytes are inhibited by gadolinium and amiloride. AB - Chondrocytes in articular cartilage utilize mechanical signals from their environment to regulate their metabolic activity. However, the sequence of events involved in the transduction of mechanical signals to a biochemical signal is not fully understood. It has been proposed that an increase in the concentration of intracellular calcium ion ([Ca2+]i) is one of the earliest events in the process of cellular mechanical signal transduction. With use of fluorescent confocal microscopy, [Ca2+]i was monitored in isolated articular chondrocytes subjected to controlled deformation with the edge of a glass micropipette. Mechanical stimulation resulted in an immediate and transient increase in [Ca2+]i. The initiation of Ca2+ waves was abolished by removing Ca2+ from the extracellular media and was significantly inhibited by the presence of gadolinium ion (10 microM) or amiloride (1 mM), which have previously been reported to block mechanosensitive ion channels. Inhibitors of intracellular Ca2+ release (dantrolene and 8-diethylaminooctyl 3,4,5-trimethoxybenzoate hydrochloride) or cytoskeletal disrupting agents (cytochalasin D and colchicine) had no significant effect on the characteristics of the Ca2+ waves. These findings suggest that a possible mechanism of Ca2+ mobilization in this case is a self-reinforcing influx of Ca2+ from the extracellular media, initiated by a Ca2+-permeable mechanosensitive ion channel. Our results indicate that a transient increase in intracellular Ca2+ concentration may be one of the earliest events involved in the response of chondrocytes to mechanical stress and support the hypothesis that deformation-induced Ca2+ waves are initiated through mechanosensitive ion channels. PMID- 10376734 TI - Monitoring glycosaminoglycan replenishment in cartilage explants with gadolinium enhanced magnetic resonance imaging. AB - We previously devised a magnetic resonance imaging method that allows for the nondestructive and quantitative determination of glycosaminoglycan concentration in excised cartilage. The technique measures the concentration of the charged contrast agent Gd-DTPA2- (gadolinium diethylenetriamine-pentaacetic acid) equilibrated within cartilage, from which the tissue distribution of glycosaminoglycan can be calculated. The goals of our study were to determine the practicality of nondestructively monitoring glycosaminoglycan concentration in cartilage explants over a long-term culture period and to determine if glycosaminoglycan could be restored to glycosaminoglycan-depleted cartilage explants maintained in long-term culture. To meet our objectives, we harvested bovine cartilage explants, treated them initially with trypsin to reduce the glycosaminoglycan concentration, and cultured them for as long as 8 weeks. Images depicting glycosaminoglycan concentration were calculated from magnetic resonance images acquired at selected intervals during the trypsinization process and the subsequent culture period. The results indicate that gadolinium-enhanced magnetic resonance imaging can follow the reduction of glycosaminoglycan concentration over the course of enzymatic digestion and the replenishment of glycosaminoglycan over several weeks of culture and that cultured cartilage explants are capable of restoring glycosaminoglycan to 85% of its initial concentration. Of particular interest, samples cultured for 5 weeks indicated a depth dependence of glycosaminoglycan regeneration to values similar to the initial physiologic distribution. Thus, this magnetic resonance imaging method may be a very powerful means for exploring the spatial and temporal evolution of glycosaminoglycan in cartilage. PMID- 10376736 TI - Effect of surface roughness and composition on costochondral chondrocytes is dependent on cell maturation state. AB - During endochondral bone formation, as occurs in fracture healing, chondrocytes are one of the first cells to see an implant surface. We tested the hypothesis that chemical composition and surface roughness affect chondrocyte differentiation, matrix synthesis, and local factor production and that the nature of the response is dependent on the state of maturation of the cells. To do this, we harvested rat growth zone and resting zone chondrocytes and examined their response to smooth and rough disk surfaces manufactured from either commercially pure titanium or titanium alloy. Profilometry, scanning electron microscopy, Auger spectroscopy, and Fourier transform infrared spectroscopy were used to characterize the surfaces. Average roughness values were 0.22 microm for smooth titanium surfaces, 0.23 microm for smooth titanium alloy surfaces, 4.24 microm for rough titanium surfaces, and 3.20 microm for rough titanium alloy surfaces. Cells were grown on the different disk surfaces until the cultures had reached confluence on plastic. The effect of the surfaces was determined by assaying cell number and [3H]thymidine incorporation as measures of cell proliferation, cell layer and cell alkaline phosphatase specific activity as markers of differentiation, and collagen production and [35S]sulfate incorporation as indicators of extracellular matrix production. In addition, the synthesis of prostaglandin E2 and transforming growth factor-beta were examined to measure changes in local factor synthesis. In growth zone and resting zone cultures, cell number and [3H]thymidine incorporation were decreased on rough surfaces; however, this effect was greater on commercially pure titanium surfaces. Cell layer and cell alkaline phosphatase specific activity were decreased in resting zone cells grown on rough surfaces. Cell alkaline phosphatase specific activity in growth zone cells was decreased on rough surfaces, whereas cell layer alkaline phosphatase specific activity was increased only in growth zone cells grown on rough commercially pure titanium surfaces. Resting zone cell collagen production was decreased only on rough commercially pure titanium, whereas in growth zone cells, collagen production was increased. Increased prostaglandin E2 release into the media was found for growth zone and resting zone cell cultures on the disks with rough surfaces. The observed effect was greater on rough commercially pure titanium. Production of transforming growth factor-beta by resting zones was similarly affected, whereas an increase in its production by growth zone cells was measured only on rough commercially pure titanium. These results indicate that surface roughness affects chondrocyte proliferation, differentiation, matrix synthesis, and local factor production and that these parameters are also affected by chemical composition. Furthermore, the nature and extent of the cell response is dependent on cell maturation. The overriding variable in response to an implant material, however, appears to be roughness of the surface. PMID- 10376735 TI - Localization and expression of cartilage oligomeric matrix protein by human rheumatoid and osteoarthritic synovium and cartilage. AB - Synovium and cartilage from patients with osteoarthritis or rheumatoid arthritis were analyzed for expression of cartilage oligomeric matrix protein. Immunostaining of synovium with antiserum to cartilage oligomeric matrix protein demonstrated positive staining in both diseases. In osteoarthritis, there was positive staining within the synovial cells and immediately subjacent connective tissue, with less intense staining in the deeper connective tissue. In rheumatoid arthritis, there was less intense staining within the synovial cells and marked intense staining in the deeper connective tissue. In situ hybridization performed with an antisense digoxigenin-labeled riboprobe to human cartilage oligomeric matrix protein confirmed the presence of cartilage oligomeric matrix protein mRNA in the cells of the synovial lining in both types of synovium. Quantitative polymerase chain reaction with a cartilage oligomeric matrix protein MIMIC demonstrated increased cartilage oligomeric matrix protein mRNA in rheumatoid cartilage and synovium as compared with osteoarthritic cartilage and synovium, respectively; mRNA levels in rheumatoid synovium were similar to those from osteoarthritic chondrocytes. As a result of the high expression of cartilage oligomeric matrix protein from rheumatoid synovium, inflammatory synovium should be considered as a potential tissue source of cartilage oligomeric matrix protein in any investigation of biological markers of cartilage metabolism. The upregulated expression of cartilage oligomeric matrix protein in inflammatory tissues suggests its in vivo regulation by cytokines. PMID- 10376737 TI - Inputs to combination-sensitive neurons of the inferior colliculus. AB - In the mustached bat, combination-sensitive neurons display integrative responses to combinations of acoustic elements in biosonar or social vocalizations. One type of combination-sensitive neuron responds to multiple harmonics of the frequency-modulated (FM) components in the sonar pulse and echo of the bat. These neurons, termed FM-FM neurons, are sensitive to the pulse-echo delay and may encode the distance of sonar targets. FM-FM neurons are common in high-frequency regions of the central nucleus of the inferior colliculus (ICC) and may be created there. If so, they must receive low-frequency inputs in addition to the expected high-frequency inputs. We placed single deposits of a tracer at FM-FM recording sites in the ICC and then analyzed retrograde labeling in the brainstem and midbrain. We were particularly interested in labeling patterns suggestive of low-frequency input to these FM-FM neurons. In most nuclei containing labeled cells, there was a single focus of labeling in regions thought to be responsive to high-frequency sounds. More complex labeling patterns were observed in three nuclei. In the anteroventral cochlear nucleus, labeling in the anterior and marginal cell divisions occurred in regions thought to respond to low-frequency sounds. This labeling comprised 6% of total brainstem labeled cells. Labeling in the intermediate nucleus of the lateral lemniscus and the magnocellular part of the ventral nucleus of the lateral lemniscus together comprised nearly 40% of all labeled cells. In both nuclei, multiple foci of labeling occurred. These different foci may represent groups of cells tuned to different frequency bands. Thus, one or more of these three nuclei may provide low-frequency input to high frequency-sensitive cells in the ICC, creating FM-FM responses. We also examined whether ICC neurons responsive to lower frequencies project to high-frequency sensitive ICC regions; only 0.15% of labeling originated from these lower frequency representations. If the spectral integration of FM-FM neurons is created at the level of the ICC, these results suggest that neurons of the anteroventral cochlear nucleus or monaural nuclei of the lateral lemniscus may provide the essential low-frequency input. In contrast, there is little evidence that the low-frequency representation of the ICC contributes to these integrative responses. PMID- 10376738 TI - Multidimensional chemotopic responses to n-aliphatic acid odorants in the rat olfactory bulb. AB - In an effort to understand the means by which similar chemical odorants are encoded in the mammalian brain, we exposed rats to a homologous series of n aliphatic acids and mapped the response of the entire olfactory bulb glomerular layer by using a high-resolution [14C]-2-deoxyglucose uptake technique. We found that these similar odorants evoked spatially clustered but distinct responses in the bulb that changed systematically with carbon chain length. In addition to these chemotopic responses, different odorants within the series evoked systematic differences along two other dimensions: amount of deoxyglucose uptake and extent of the glomerular layer showing high activity. Increases along these two dimensions also were correlated with increasing carbon number. The focal glomerular responses were mirrored by responses in deeper bulb layers. Decreasing the odorant concentration decreased the deoxyglucose uptake within focal regions. The focal regions of activity occurred in pairs involving both medial and lateral representations in the bulb, perhaps reflecting the paired medial and lateral projections of olfactory sensory neurons expressing specific types of odorant feature receptor proteins. The observed spatial pattern of response also may explain both the failure of some bulb lesions to interfere with behavioral olfactory responses and the success of other lesions in blocking olfactory responses. These data support a model of parallel, distributed processing of odorants along multiple dimensions. They also support the notion that analyses of the spatial relationships among odorant responses in the olfactory bulb can demonstrate aspects of the mechanism for odor chemical coding. PMID- 10376739 TI - Development of metabotropic glutamate receptors from trigeminal nuclei to barrel cortex in postnatal mouse. AB - Expression patterns of group I (mGluR1alpha and mGluR5) and group II (mGluR2/3) metabotropic glutamate receptor subtypes were examined immunocytochemically in the trigeminal system of mice during the first 3 weeks of postnatal development, when somatotopic whisker representations are sequentially established from brainstem through thalamus to cerebral cortex. Immunostaining for all three epitopes formed whisker-related patterns in the trigeminal nuclei from postnatal day (P) 0, in the ventral posterior thalamic nucleus from P2, and in the posteromedial barrel subfield of somatosensory cortex (SI) from P4. The appearance of whisker-related patterns was preceded by increased levels of immunostaining of the neuropil, which subsequently declined from the trigeminal nuclei upward. In SI, mGluR1alpha-positive neurons were observed in all cortical layers from P2. mGluR5 was localized in neurons, glial cells, and neuropil from P2. mGluR2/3 immunostaining was distributed only in the neuropil at all ages. The three receptor subtypes showed moderate to high expression in deep layer V throughout development. Transient expression peaked in the hollows of layer IV barrels from P4 to P9, and then fell off as expression increased in supragranular layers from P14 to P21. The deep aspect of the cortical subplate (layer VIb) showed dense mGluR5 and less dense mGluR1alpha immunostaining throughout development. Up-regulation of expression of group I and II mGluRs is correlated with the growth and refinement of connectivity and the establishment of somatotopic patterns in the three main relay stations of the trigeminal system. This finding suggests roles for mGluRs in the early processing of sensory information and in developmental plasticity. PMID- 10376740 TI - Brain weight throughout the life span of the chimpanzee. AB - Studies on human postmortem material report lower brain weights in older than in younger cohorts, whereas there is no apparent change with age in the rhesus monkey. In view of these contrasting results, we examined the pattern of brain weight across the life span in the chimpanzee, one of the closest biological relatives of humans. To place the study in context of the empirical life expectancy of the chimpanzee, we first performed a survival analysis on data from 275 chimpanzees that were maintained in the colony of the Yerkes Primate Center. The survival analysis revealed the maximum life spans of female and male chimpanzees to be about 59 and 45 years, respectively. We examined fresh brain weights from 76 chimpanzees ranging in age from birth to 59.4 years of age. The brains were taken from 9 infants (birth to 1 year of age), 25 juveniles (1-7 years), 13 adolescents (7-15 years), 21 young adults (15-30 years), and 8 old adults (over 30 years). Adult brain weight was achieved by the age of 7 years. The adolescent and young adult chimpanzees had the largest brain weights; in these two age groups combined, the mean brain weight (+/- standard deviation) was 368.1 g (+/-37.3) for females (n = 17) and 405.6 g (+/-39.4) for males (n = 17). This sex difference was statistically significant (P < 0.01). Simple linear regression performed on the combined material from females and males aged 7 years and older revealed a decline in brain weight with advancing age of 1.1 g/year (P < 0.05). When the effect of sex on brain weight was statistically controlled for, the loss of brain weight with age was 0.9 g/year (P = 0.07). These results suggest that brain weight declines moderately with age in the chimpanzee as it does in humans. PMID- 10376741 TI - Relationships between cytochrome oxidase (CO) blobs in primate primary visual cortex (V1) and the distribution of neurons projecting to the middle temporal area (MT). AB - The cytochrome oxidase (CO) blobs and interblobs in layer 3B of primate visual cortex have different sets of corticocortical connections. Cortical layers below layer 3B also project corticocortically, but the relationship of efferent projections from the deeper layers to the overlying blob/interblob architecture is less clear. We studied the tangential organization of neurons projecting from primary visual cortex (V1) to the middle temporal visual area (MT) and their relationship to the CO blobs. MT-projecting neurons in two primate species, bush babies and owl monkeys, were retrogradely labeled, then charted in tangential sections, and compared to the positions of the overlying CO blobs. In both primate species, MT-projecting neurons in layer 3C were unevenly distributed in the tangential plane, with dense patches of labeled cells that were aligned with the CO blobs. A novel two-dimensional spatial correlation method was used to show the colocalization of MT-projecting cells with the overlying blobs. Chi-square analyses performed with the cortical surface equally divided into compartments of blob, interblob, and blob/interblob borders showed that blob columns tended to have about 1.5 times more MT-projecting cells (P < 0.0001) than interblob columns. Similar analyses were applied to published data on V1 cells projecting to area MT in macaque monkey (Shipp and Zeki [1989] Euro J Neurosci 1:310-332). Again, the results showed a significant correlation between the cell distribution and CO blobs. Taken together, these results suggest that layer 3C is not uniform but is made up of a mosaic of cells that project to area MT and cells that project to some other location. These findings also indicate that the mosaic organization of layer 3C is related in some unique way to the overlying CO architecture. PMID- 10376742 TI - Reduction in cell size during development of the spinal cord. AB - During development, spinal cord cells of the frog Xenopus laeuis undergo a reduction in size. This phenomenon occurs during neural tube formation and continues at least until the start of metamorphosis. The number and shape of spinal cord cells also changes, but not always in synchrony with the reduction in cell size. The concomitant change in size and number of spinal cord cells during embryogenesis suggests that a cleavage type of reductive division contributes to the decrease in cell size. Blocking cell division with a combination of hydroxyurea and aphidicolin (HUA) stops the decrease in cell size during embryonic development without affecting the differentiation of a specific class of catecholaminergic neurons. HUA treatment during larval stages does not block the decrease in catecholaminergic neuron size. Thus, both mitotic and postmitotic cells decrease in size during spinal cord development. The two mechanisms are prevalent at different developmental stages with reductive division and cellular atrophy common during embryonic and larval phases, respectively. Like other regressive changes such as cell death and synapse elimination, decreases in cell size affect spinal cord morphogenesis and presumably the function of developing spinal cord cells. PMID- 10376743 TI - Organization of olfactory and multimodal afferent neurons supplying the calyx and pedunculus of the cockroach mushroom bodies. AB - The mushroom bodies of neopteran insects are considered to be higher olfactory centers because their calyces receive abundant collaterals of projection neurons from the antennal lobes. However, intracellular recordings of mushroom body efferent neurons demonstrate that they respond to multimodal stimuli, implying that the mushroom bodies receive a variety of sensory cues. The present account describes new features of the organization of afferent neurons supplying the calyces of the cockroach Periplaneta americana. Afferent terminals segment the calyces into discrete zones, I, II, III, and IIIA, which receive afferents from 1) two discrete populations of sexually isomorphic olfactory glomeruli, 2) two types of male-specific olfactory glomeruli, 3) the optic lobes, and 4) multimodal interneurons that originate in protocerebral neuropils. In addition, intracellular recordings and dye fills show that at least four morphologically distinct GABAergic elements link many regions of the protocerebrum to the calyces. A new type of touch-sensitive centrifugal neuron has been identified terminating in the pedunculus. The dendrites of this afferent reside in satellite neuropil, beneath the mushroom body's medial lobe, which is supplied by collaterals from medial lobe efferent neurons and by terminals from the central complex. The role of this centrifugal cell in odorant sampling is considered. Golgi impregnation identifies other afferents in proximal regions of the calyx (zone IIIA) that also originate from satellite neuropils, suggesting major reafference from the medial lobes channeled through this region. The relevance of multimodal supply to the calyx in odorant discrimination is discussed as are comparisons between mushroom body organization in this phylogenetically basal neopteran and other taxa. PMID- 10376744 TI - Representation of the calyces in the medial and vertical lobes of cockroach mushroom bodies. AB - Previous studies of honey bee and cockroach mushroom bodies have proposed that afferent terminals and intrinsic neurons (Kenyon cells) in the calyces are arranged according to polar coordinates. It has been suggested that there is a transformation by Kenyon cell axons of the polar arrangements of their dendrites in the calyces to laminar arrangements of their terminals in the lobes. Findings presented here show that cellular organization in the calyx of an evolutionarily basal neopteran, Periplaneta americana, is instead rectilinear, as it is in the lobes. It is shown that each calyx is divided into two halves (hemicalyces), each supplied by its own set of Kenyon cells. Each calyx is separately represented in the medial lobe where the dendritic trees of some efferent neurons receive inputs from one calyx only. Kenyon cell dendrites are arranged as narrow elongated fields, organized as rows in each hemicalyx. Dendritic fields arise from 14 to 16 sheets of Kenyon cell axons stacked on top of each other lining the inner surface of the calyx cup. A sheet consists of approximately 60 small bundles, each containing 5-15 axons that converge from the rim of the calyx to its neck. Each sheet contributes to a pair oflaminae, one dark one pale, called a doublet, that extends through the mushroom body. Dark laminae contain Kenyon cell axons packed with synaptic vesicles. Axons in pale laminae are sparsely equipped with vesicles. By analogy with photoreceptors, and with reference to field potential recordings, it is speculated that dark laminae are continuously active, being modulated by odor stimuli, whereas pale laminae are intermittently activated. Timm's silver staining and immunocytology reveal a second type of longitudinal division of the lobes. Five layers extend through the pedunculus and lobes, each composed of subsets of doublets. Four layers represent zones of afferent endings in the calyces. A fifth (the y layer) represents a specific type of Kenyon cell. It is concluded that the mushroom bodies comprise two independent modular systems, doublets and layers. Developmental studies show that new doublets are added at each instar to layers that are already present early in second instar nymphs. There are profound similarities between the mushroom bodies of Periplaneta, an evolutionarily basal taxon, and those of Drosophila melanogaster and the honey bee. PMID- 10376745 TI - Multimodal efferent and recurrent neurons in the medial lobes of cockroach mushroom bodies. AB - Previous electrophysiological studies of cockroach mushroom bodies demonstrated the sensitivity of efferent neurons to multimodal stimuli. The present account describes the morphology and physiology of several types of efferent neurons with dendrites in the medial lobes. In general, efferent neurons respond to a variety of modalities in a context-specific manner, responding to specific combinations or specific sequences of multimodal stimuli. Efferent neurons that show endogenous activity have dendritic specializations that extend to laminae of Kenyon cell axons equipped with many synaptic vesicles, termed "dark" laminae. Efferent neurons that are active only during stimulation have dendritic specializations that branch mainly among Kenyon cell axons having few vesicles and forming the "pale" laminae. A new category of "recurrent" efferent neuron has been identified that provides feedback or feedforward connections between different parts of the mushroom body. Some of these neurons are immunopositive to antibodies raised against the inhibitory transmitter gamma-aminobutyric acid. Feedback pathways to the calyces arise from satellite neuropils adjacent to the medial lobes, which receive axon collaterals of efferent neurons. Efferent neurons are uniquely identifiable. Each morphological type occurs at the same location in the mushroom bodies of different individuals. Medial lobe efferent neurons terminate in the lateral protocerebrum among the endings of antennal lobe projection neurons. It is suggested that information about the sensory context of olfactory (or other) stimuli is relayed by efferent neurons to the lateral protocerebrum where it is integrated with information about odors relayed by antennal lobe projection neurons. PMID- 10376746 TI - Development of GABA, glycine, and their receptors in the auditory brainstem of gerbil: a light and electron microscopic study. AB - Inhibitory synaptic transmission is known to play an important role during the maturation of central auditory pathways. While there is a lot of information on the modulatory role of glycine (Gly) on the postsynaptic target nuclei in the developing auditory brain stem, such a role for gamma-aminobutyric acid (GABA) in the lateral superior olive (LSO) of neonatal gerbil has been only recently reported (Kotak and Sanes [1997] Soc Neurosci Abst 23:1549; Kotak et al. [1998] J Neurosci 18:4646-4655). Here we present further immunohistochemical findings and the first ultrastructural evidence documenting a significant decrease in the postsynaptic localization of the beta2,3 subunit of the GABA(A) receptor from postnatal day (P)4 to P14 in the LSO of gerbil and the shift in the location of most of the staining from dendritic to astroglial over the same time course. There was a concomitant increase in staining for the Gly receptor (GlyR) anchoring protein, gephyrin. At the same time, GABA and Gly did not show a significant change in their staining pattern, suggesting that the transmitter levels are not particularly indicative of the inhibitory function in the neonatal gerbil LSO, but their receptors on the postsynaptic cells are. The observations of the present study suggest that the early GABAergic inhibition may be important in establishing appropriate synaptic contacts in the LSO of gerbil. PMID- 10376747 TI - A prospective study on quality of life of laryngeal cancer patients treated with radiotherapy. AB - BACKGROUND: The aim of this study was to describe prospectively quality of life and mood before and after radiotherapy for laryngeal cancer. METHODS: Sixty-five patients with Tis-T3 laryngeal cancer treated with radiotherapy completed the European Organization for Research and Treatment of Cancer (EORTC) Core Questionnaire, the EORTC Head and Neck Cancer module, and the Center for Epidemiologic Studies Depression Scale before treatment and 6 and 12 months later. RESULTS: There was a significant but temporary deterioration of physical functioning, fatigue and most head and neck symptoms. Speech was the only symptom which improved. Patients with T2 tumors had significantly worse physical symptoms compared with patients with T1 tumors. There was a high level of depressive symptomatology at baseline, followed by an improvement after treatment. CONCLUSIONS: After radiotherapy for laryngeal cancer, a temporary deterioration of physical functioning and symptoms occurs, mostly caused by side effects of treatment. Despite physical deterioration, there is an improvement of emotional functioning and mood after treatment, probably as a result of psychological adaptation and coping processes. PMID- 10376748 TI - National Cancer Data Base report on cancer of the head and neck: acinic cell carcinoma. AB - BACKGROUND: Management of acinic cell carcinoma is based on reports of small numbers of cases accrued over several decades. METHODS: The National Cancer Data Base (NCDB) identified 1353 cases of acinic cell carcinoma of the head and neck for the years 1985 to 1995. Chi square analyses of selected contingency tables and Wilcoxon regression analyses of selected survival stratifications are presented. RESULTS: Five-year survival was 83.3% (observed) and 91.4% (disease specific). Worse survival was associated with high grade (p < .0001), age greater or equal to 30 years (p = .0055), and the presence of metastatic disease (p < .0001). CONCLUSIONS: An aggressive subset of acinic cell carcinoma which is characterized by high grade and advanced stage rarely occurs in patients younger than 30 years old. Although better outcome was not statistically demonstrated for combined therapy, surgery with irradiation is the most common management in the United States for cases with regional metastases, high grade, and microscopic positive margins. PMID- 10376749 TI - Laser treatment of symptomatic anterior pharyngeal pouches after laryngectomy. AB - BACKGROUND. An anterior pharyngeal pouch is a common anatomic variation after total laryngectomy. The exact part the pouch has to play in postlaryngectomy swallowing disorders is unknown but might be bigger than expected. METHODS: Nine patients with a symptomatic pouch were treated with CO2 laser and questioned about pre- and postoperative complaints. RESULTS: Eight out of nine patients noticed an obvious improvement of their swallowing complaints after laser surgery. CONCLUSION: Based on the results, it is concluded that CO2-laser treatment with division of the bar of tissue posterior to the pouch is seen as a safe and simple way to treat patients for symptoms that can be wrongly attributed to a total laryngectomy. PMID- 10376750 TI - Local radiation dose, fixation, and non-union of mandibulotomies. AB - BACKGROUND. Non-union of mandibulotomies is a serious complication but risk factors are unclear. No data are available on the effect of the specific dose of irradiation at the osteotomy site. METHODS: Forty-eight patients (36 men, 12 women, median age 58 years) undergoing a mandibulotomy and postoperative radiotherapy were enrolled in the study. Twenty-five had wire and 23 had plate osteosyntheses. Five patients had a non-union and three had minor complications. The distribution of the radiation dose and the specific dose at the mandibulotomy site were calculated. Parametric tests and logistic regression were used for analysis. RESULTS: Non-union was not related to a specific dose of irradiation at the osteotomy site. Also, no effect could be attributed to type of fixation. CONCLUSIONS: There is no evidence for a single factor, radiation, or fixation type, that contributes to a non-union. We recommend a careful surgical technique. PMID- 10376751 TI - Quality of life after laryngectomy: are functional disabilities important? AB - BACKGROUND: The purpose of this study was to determine the functional disabilities and overall quality of life (QOL) of patients successfully treated (ie, without evidence of disease at two years) for laryngeal or hypopharyngeal cancer by a total laryngectomy. METHODS: The University of Washington QOL questionnaire was administered to 10 patients prior to laryngectomy, at one year postlaryngectomy, and at two years postlaryngectomy. RESULTS: Postlaryngectomy QOL scores were not significantly different from prelaryngectomy scores. In all QOL domains the severity of functional disability was not significantly correlated with its importance. Ninety percent of patients (9/10) reported that compared with one year prior to the diagnosis of cancer their general health was the same or better at two years postlaryngectomy. Seventy percent of patients (7/10) reported having a good to excellent overall QOL. CONCLUSIONS: Though the loss of voice is disabling, the functional limitations caused by a laryngectomy do not necessarily translate into a worse overall QOL. Future research is needed to determine whether the importance of individual QOL domains changes as patients adjust to the experience of having and surviving cancer. PMID- 10376752 TI - Fatty acid synthase expression is increased in neoplastic lesions of the oral tongue. AB - BACKGROUND: Fatty acid synthase (FASE) is required for fatty acid synthesis. Elevated levels of FASE have been observed in a variety of malignancies. METHODS: We examined the expression of FASE in 56 primary squamous cell carcinomas (SCC) of the tongue using immunohistochemistry (IHC) with a monoclonal antibody to FASE. RESULTS: Immunoreactivity was low in histologically normal epithelium (0.42 +/- .07, n = 43), moderate in mildly dysplastic epithelium (1.41 +/- 11, n = 40), and strong in SCC of the tongue (1.64 +/- 10, n = 50). Both mild dysplasia and SCC stained more strongly than histologically normal epithelium (p<0.00001). Well differentiated tumors showed increased immunoreactivity when compared to less well-differentiated tumors (p=0.044). Decreased overall survival was observed among patients with tumors with low immunoreactivity (p = 0.04). CONCLUSIONS: Increased expression of FASE in dysplasia and squamous carcinomas of the oral tongue may be an indicator of both differentiation and early neoplastic change. FASE expression may be useful diagnostically, prognostically, and as a potential target for therapy. PMID- 10376753 TI - Immunohistochemical study of sialyl Le(a) and sialyl Le(x) antigen in oral squamous cell carcinoma: the association of sialyl Le(a) expression with the metastatic potential. AB - BACKGROUND: Carbohydrate antigens in cancer cells are considered to be involved in the binding of cancer cells to the endothelium during metastasis. METHODS: Seventy cases of primary oral squamous cell carcinoma (SCC) were obtained from biopsy specimens and were analyzed immunohistochemically using an antibody against sialyl Lewis (Le)a or sialyl Le(x). Flow cytometry was performed to detect the sialyl Le(a) or sialyl Le(x) expressed on oral SCC cell lines. RESULTS: The expressions of sialyl Le(a), but not sialyl Le(x), of primary tumors significantly correlated to nodal metastasis; 71% of the metastatic cases express sialyl Le(a) and the cases with positive sialyl Le(a) and no sialyl Le(x) demonstrated a high incidence of metastasis (80%). A flow cytometric study demonstrated the oral SCC cell line, which can metastasize in nude mice, to express a high level of sialyl Le(a). CONCLUSION: The high expression of sialyl Le(a) in primary tumors may thus be involved in nodal metastasis and therefore predict a poor prognosis in oral SCC. PMID- 10376754 TI - Fractionated stereotactic radiation therapy for locally recurrent nasopharynx cancer: report of three cases. AB - BACKGROUND: This article reports on experience with fractionated stereotactic radiation therapy (FSRT) for locally recurrent nasopharynx cancer. METHODS: Three patients with locally recurrent nasopharynx cancer were given FSRT as reirradiation between September 1995 and August 1996. Application of FSRT was the third radiation therapy in two patients. Authors used the individually made relocatable Gill-Thomas-Cosman (GTC) stereotactic frame, and the radiation dose planning was performed using XKnife-3. The total doses to the recurrent tumor were 45 Gy/18 fractions in two patients, who were given concurrent chemotherapy as a radiosensitizer, and 50 Gy/20 fractions in the other patient. In all three patients the dose per fraction was 2.5 Gy, and the fraction schedule was to give five daily treatments per week. RESULTS: Authors observed satisfactory symptomatic improvement and remarkable objective tumor size decrease through the magnetic resonance (MR) images taken one month post-FSRT in all three patients. No neurological side effect was observed. All three patients died with regional and distant seeding outside the FSRT field at seven, nine, and nine months, respectively. CONCLUSION: FSRT as reirradiation for locally recurrent nasopharynx cancer seemed to be effective and safe. PMID- 10376755 TI - Radiation induced sarcoma of the head and neck. AB - BACKGROUND: Radiation-induced sarcoma of the head and neck (RISHN) is a long-term complication of treatment. The rarity of this tumour is reflected in the very few series reported in the English language medical literature. The incidence of RISHN is, however, likely to increase due to progressive aging of the population combined with improved survival in head and neck cancer patients resulting from better treatment regimes. Diagnosis and management of this problem can be extremely challenging and the overall outlook has been reported to be very bleak. As survival data from isolated case reports cannot be expected to provide reliable information on outcome, we have reviewed 69 cases reported in the English medical literature since 1966 and pooled this information with our experience in treatment of RISHN. PATIENTS AND METHODS: Ten patients with features of a RISHN were treated at the Royal Marsden Hospital between 1944 and 1997. The features of RISHN, treatment, and outcome were analysed in these patients. Additionally, 61 eligible patients with RISHN reported in the literature between 1966 and 1997 were pooled with nine of our patients to form the RISHN group (n = 70). This group was then compared for survival with 124 patients with a diagnosis of head and neck sarcoma registered on the Head and Neck Sarcoma database at the Royal Marsden Hospital (SHN group). Lifetables were constructed using the Kaplan-Meier method and compared using the log-rank test. RESULTS: There was no site of predilection for RISHN, but malignant fibrous histiocytoma (MFH) was the commonest pathological diagnosis. The period of latency between initial radiation therapy and diagnosis of RISHN ranged from 9 to 45 years with a median of 17 years. Surgery was the mainstay of treatment and follow-up ranged from 6 months to 15 years with a median of 48 months. The actuarial five-year disease free survival in these patients was 60%. CONCLUSION: There is at present little or no prospect for effective prevention of RISHN and therefore, a high index of suspicion based on the patient's symptoms assumes great importance in the outcome of these patients. Although surgical management of RISHN is challenging because of the close proximity of the tumour to important regional structures and the technical difficulties of operating in an irradiated area, complete surgical excision appears to offer the best means for palliation and the only realistic chance for long-term survival. PMID- 10376756 TI - The pathogenesis of vascular thrombosis and its impact in microvascular surgery. AB - As the use of free tissue transfer becomes more wide-spread, it is important for both the ablative surgeon and the reconstructive surgeon to understand the factors that contribute to successful revascularized tissue transfer. The purpose of this two part review is to provide a basic science overview of the problem of failed free tissue transfers. The first part will focus on the pathogenesis of thrombosis at the anastomotic site, and part two will discuss the pathogenesis of the no-reflow phenomenon. The pathophysiology and therapeutic interventions to prevent and treat anastomotic thrombosis and the no-reflow phenomenon will be discussed. PMID- 10376757 TI - Primary placement of a voice prosthesis on transposed colon after total pharyngolaryngoesophagectomy. AB - BACKGROUND: Primary placement of a voice prosthesis may aid rehabilitation after total laryngectomy. METHODS: We present a rare clinical situation of a T4 NO MO squamous cell carcinoma of the hypopharynx and esophagus in a patient who had previously undergone a transmesocolic Billroth II gastrectomy. RESULTS: The patient benefited from a total pharyngolaryngoesophagectomy, with reconstruction using a transverse-descending colon transposition, and primary placement of a low pressure voice prosthesis. CONCLUSION: Primary placement of a voice prosthesis may be successful even in a patient who requires extensive pharyngoesophageal reconstruction using transposed colon. To our knowledge, there has been no previous report of primary placement of a voice prosthesis on a colon autograft. PMID- 10376758 TI - Vascular transformation of sinuses in bilateral cervical lymph nodes. AB - BACKGROUND: Vascular transformation of sinuses (VTS) in lymph node has been infrequently reported. We present a case of incidental VTS in bilateral cervical nodes discovered at the time of operation for tongue cancer. METHODS: Standard histopathologic review was undertaken. RESULTS: In this case, a spectrum of varied vasoformative patterns involving lymph nodes in either a pan-nodal or localized fashion were identified. The hilum was occupied by smooth muscle proliferation and adipose tissue. Two nodes also displayed similar features of angiolipomatous hamartoma. No metastatic carcinoma was found in lymph nodes with VTS. CONCLUSIONS: This case suggests that lymphovenous congestion and distention represented the major cause through which VTS developed. PMID- 10376759 TI - Cervical cystic lymph node metastasis as first manifestation of occult papillary thyroid carcinoma: report of seven cases. AB - BACKGROUND: Cervical cystic lymph node metastases as first and sole manifestation of occult papillary thyroid carcinoma are observed exceptionally rarely. In the seven patients here reported, a cystic, ovoid mass in the lateral aspect of the neck was the initial symptom of the papillary microcarcinoma. METHODS: There were six men and one woman, aged between 17 and 54 years (mean 31.7 years), who complained of round, movable, painless masses in the lateral aspect of the neck. Two patients were first seen with two cervical tumors. Tumors had been present from a few days to 12 months (mean 5.1 months). The cystic nature of tumors was demonstrated by echographic studies. In all cases, thyroid tumors were not palpable on physical examination and no abnormalities of the thyroid gland were shown by other diagnostic procedures. RESULTS: The diagnosis was made preoperatively by fineneedle aspiration cytology of the nodes in five of the seven cases. All patients underwent thyroidectomy with conservative neck dissection followed by radioactive iodine therapy. After a follow-up period from 1 to 7 years, all patients are alive with no apparent signs of recurrence or metastasis. CONCLUSIONS: In a young patient with solitary lateral cervical cyst, the diagnosis of lymph node metastasis from occult papillary thyroid carcinoma should be considered. Any lateral mass requires tissue diagnosis, and fine-needle aspiration is usually adequate for clarification of the histology. Ipsilateral modified neck dissection and total thyroidectomy followed by radioactive iodine therapy offers a favorable prognosis. PMID- 10376760 TI - Estimation of cytochrome P-450 CYP1A2 activity in 863 healthy Caucasians using a saliva-based caffeine test. AB - A pronounced variability limits the usefulness of CYP1A2 phenotyping for drug therapy, for evaluating liver function, and for assessing the role of this enzyme in carcinogenesis. To identify and quantify sources of this variation, we estimated CYP1A2 activity in 863 healthy Caucasians using caffeine clearance derived from saliva concentrations before and 5-7 h after a caffeine test dose. Data from 786 individuals were eligible for evaluation (mean age 39 years, 415 women including 94 taking oral contraceptives, 401 non-smokers). Overall geometric mean (geometric SD) caffeine clearance was 1.34 ml min(-1) kg b.w.(-1) (1.65). The effect of the following covariates was evaluated by analysis of covariance: age, sex, oral contraceptives, body height, body weight, body mass index, number of cigarettes smoked, tar exposure from smoking, several indices of dietary caffeine consumption, intake of sauerkraut, and country of residence (Germany, Bulgaria or Slovakia). Estimated changes relative to arbitrarily defined basal caffeine clearance (male, non-smoking, German resident) exerted by significant (P < 0.05) covariates were: coffee, 1.45-fold per litre of coffee drunk daily; body mass index, 0.99-fold per kg m(-2); smoking, 1.22-fold, 1.47 fold, 1.66-fold, and 1.72-fold for 1-5, 6-10, 11-20, and > 20 cigarettes smoked per day, respectively; oral contraceptives, 0.72-fold; country of residence, 0.81 fold and 0.74-fold for Bulgaria and Slovakia, respectively; female, 0.90-fold. These covariates explained 37% of overall variation. The 95% confidence interval of individual clearance was 0.46-2.20 times the predicted value. No relevant polymorphism was found for CYP1A2 activity when adjusted for covariate effects. PMID- 10376761 TI - Four amino acid changes are associated with the Aldh3a1 locus polymorphism in mice which may be responsible for corneal sensitivity to ultraviolet light. AB - We studied the phenotype and the nucleotide sequence for the cDNAs of the Aldh3a1a and Aldh3a1c allelic forms of the dioxin-inducible cytosolic aldehyde dehydrogenase (ALDH3A1) present in inbred mouse strains. This gene is constitutively expressed in cornea, stomach, skin, urinary bladder and lungs. The Aldh3a1a allele is found in most inbred mouse strains and codes for a 'high activity' corneal enzyme compared to the 'low-activity' encoded by the Aldh3a1c allele in SWR/J strain. The 'low-activity' variant is associated with extensive corneal clouding after a single exposure to ultraviolet light. The ALDH3A1 phenotype was examined in tissues from inbred mouse strains carrying the Aldh3a1a allele including, SJL/J, C57BL/6 J/Ibg, DBA/2 J/Ibg, C3H/Ibg and the Aldh3a1c allele (SWR/J). Only trace levels of ALDH3A1 activity were found in all SWR/J tissues. All other strains had significant levels of ALDH3A1 activity in eye, stomach, skin, less in urinary bladder and lungs and only trace amounts in liver. However, no differences were found in corneal and stomach ALDH3A1 mRNA levels between the 'low-' and 'high-activity' variants. A 1556-bp ALDH3A1 cDNA fragment, containing the entire coding region plus 5' and 3' untranslated regions, was amplified by reverse transcriptase-polymerase chain reaction from SWR/J and DBA/2 J/Ibg mouse strains. Sequence analysis revealed 13 nucleotide changes in the Aldh3a1c allele. Four of these changes result in G88R, I154N, H305R and I352V substitutions, whereas nine changes are silent. The I154N disrupts a potential alpha helix, which belongs to the Rossmann fold. Replacement of Arg with the more ionizable His at position 305 of a beta strand might directly affect catalytic activity of the enzyme. It is likely that structural changes associated with these amino acid changes are responsible for the loss of ALDH3A1 enzymatic activity in SWR/J mice. PMID- 10376762 TI - Phenotyping of flavin-containing monooxygenase using caffeine metabolism and genotyping of FMO3 gene in a Korean population. AB - Flavin-containing monooxygenase (FMO) activity was determined in 82 Korean volunteers by taking molar concentration ratio of theobromine and caffeine present in the 1 h urine (between 4 and 5 h) samples collected after administration of a cup of coffee containing 110 mg of caffeine. Among 82 volunteers, there were 19 women and 63 men (30 smokers and 52 non-smokers). Volunteers were divided into two groups comprising low (0.53-2.99) and high (3.18 11.95) FMO activities separated by an antimode of 3.18. Peripheral bloods were sampled from these volunteers and their genomic DNAs were amplified by polymerase chain reaction with oligonucleotides designed from intronic sequences of human FMO3 gene. Comparing nucleotide sequences of the amplified FMO3 gene originating from randomly selected individuals with low and high FMO activities, nine point mutations were identified in the open reading frame sequences. Among these nine mutations, three FMO3 mutant types (FMO3/Stop148, Lys158 and Gly308) were selected and correlated with FMO activities observed in our Korean population. A rare FMO3/Stop148 mutant allele originating from FMO3/Gly148 occurred by substitution of G442T in exon 4 and yielded a premature TGA stop codon. The stop codon was detected in one individual having the second lowest FMO activity and he had the mutation in heterozygous state. In a pedigree study, he was found to have inherited the mutation from his mother who also had a heterozygous stop codon and equally low FMO activity. In our volunteers, two other common mutations were detected in exons 4 and 7. The one in exon 4 resulted from a G472A change eliminating a HinfI restriction site and produced an amino acid substitution from Glu158 to Lys. The other mutation in exon 7 resulted from an A923G change generating a DraII restriction site and produced a non-conservative replacement of Glu308 to Gly. Based on the secondary structure maps of FMO3 enzyme proteins for these two mutant types, FMO3/Gly308 mutation transformed the helix structure into a sheet shape and indicated that dysfunctional FMO3 may be produced. FMO3/Lys158 mutation did not alter the secondary structure. Approximately 80% of volunteers with homozygous and/or heterozygous mutations on either one or two of these mutations had low FMO activities. Thus, individuals with these FMO3 gene mutations may have defective metabolic activity for many clinically used drugs and dietary plant alkaloids which are oxidized primarily by hepatic FMO3. PMID- 10376763 TI - Human glutathione S-transferase P1 polymorphism and susceptibility to smoking related epithelial cancer; oral, lung, gastric, colorectal and urothelial cancer. AB - The A/G polymorphism at nucleotide 313 in the glutathione S-transferase P1-1 (GSTP1) gene was examined in patients with different types of smoking-related cancers (oral, lung, gastric, colorectal and urothelial cancers) and healthy control individuals. This polymorphism results in an amino acid substitution from isoleucine to valine at residue 105, which reduces catalytic activity of the enzyme. In control individuals, 23.8% of individuals had GSTP1 AG or GG genotype. This rose to 37.3% [n = 83, odds ratio = 1.93 (1.05-3.58), P = 0.035] in oral cancer patients. No increase in the frequency of the GSTP1 AG or GG genotype was obtained in lung, gastric, colorectal or urothelial cancers in this Japanese population. After grouping by smoking status, no consistent difference was observed between smoking patients and corresponding control individuals for the frequency of the GSTP1 A/G polymorphism for any cancer. However, a moderate risk (odds ratio = 2.78; 95% confidence interval 1.06-7.51) was associated with this polymorphism in the non-smoking group of oral cancer patients. The results suggest the GSTP1 polymorphism at nucleotide 313 may be associated with susceptibility to oral squamous cell carcinoma in the Japanese population. PMID- 10376764 TI - Genetic differences in spatial learning between Dark Agouti and Sprague-Dawley strains: possible correlation with the CYP2D2 polymorphism in rats treated neonatally with methamphetamine. AB - Following neonatal exposure to d-methamphetamine, adult rats have previously been shown to exhibit augmented acoustic startle and spatial learning deficits. d Methamphetamine is structurally similar to several phenylethylamines that are metabolized by CYP2D6. In humans, allelic differences in the CYP2D6 confer the extensive or poor metabolizer phenotype for the more than three dozen drugs that are members of the CYP2D6-mediated 'debrisoquine/sparteine panel.' An analogous genotype exists with the CYP2D2 gene in rats. Female Dark Agouti rats show the poor metabolizer phenotype, whereas Sprague-Dawley rats show the extensive metabolizer phenotype; male Dark Agouti rats are intermediate. We sought to test the possibility that these strains might exhibit altered d-methamphetamine induced developmental neurotoxicity. Dark Agouti and Sprague-Dawley litters (11 20 days of age) were given d-methamphetamine or vehicle alone subcutaneously twice daily (15 mg/kg). Offspring were assessed as adults (beginning at 50 days of age) on acoustic startle, straight-channel swimming, and spatial learning and memory in a Morris hidden platform maze. Increases in d-methamphetamine-induced acoustic startle were found in both male and female Dark Agouti rats, but not Sprague-Dawley rats. In the Morris maze, d-methamphetamine-induced spatial navigation deficits were found in both strains among males, suggesting some mechanism other than the CYP2D2 polymorphism. In contrast, among females only the d-methamphetamine-treated Dark Agouti rats showed deficits in spatial navigation. The maze deficits in Dark Agouti females, and enhanced acoustic startle in Dark Agouti females and males, support the hypothesis that the CYP2D2 poor metabolizer phenotype confers increased vulnerability to d-methamphetamine-induced developmental neurotoxicity, indicating that the parent drug rather than a CYP2D2 mediated metabolite is responsible for this behavioural defect--which occurs in adults who had been exposed to d-methamphetamine during the neonatal period. PMID- 10376765 TI - Low levels of p53 are associated with resistance to tetrachlorodibenzo-p-dioxin toxicity in DBA/2 mice. AB - We show here that DBA/2 strain mice have a complex mutation/polymorphism in the promoter region of the Trp53 locus (the mouse p53 locus). This region has previously been shown to be essential for p53 expression. We further show that the DBA/2 mutation is associated with approximately fourfold lower p53 levels in thymocytes treated with the DNA-damaging agent etoposide in-vitro, and with relative resistance of these thymocytes to apoptosis induced by the DNA-damaging agent etoposide compared with C57BL/6 mice. When part of the promoter containing this mutation was inserted into a plasmid containing a luciferase reporter gene but lacking eukaryote promoter sequences and transfected into MCF-7 human breast cell line cells, the mean luciferase activity was slightly less with the DBA/2 than with the C57BL/6 promoter-reporter construct (p < 0.01). We found that DBA/2xC57BL/6 F2 hybrid mice with the DBA/2 genotype at the Trp53 locus were relatively resistant to tetrachlorodibenzo-p-dioxin toxicity, and this resistance was additive with resistance associated with the Ahr locus. DBA/2 mice are long lived and do not have particularly high levels of cancer, suggesting either that they carry other compensatory tumour resistance alleles (such as Ahr(d)), or that, while there may be a p53 protein dosage effect for acute toxicity, lower than normal levels of p53 may still be sufficient to protect against cancer. In evolutionary terms, it may be better to maintain low levels of p53 in order to avoid death from acute toxicity, even at the expense of a higher incidence of cancer in later life. PMID- 10376766 TI - Characterization of a variable number tandem repeat region in the thiopurine S methyltransferase gene promoter. AB - Characterization of the genetic polymorphism of thiopurine S-methyltransferase enzyme (TPMT; EC 2.1.1.67) is required because of its clinical importance for patients exposed to thiopurine drugs. A number of point mutations have already been characterized in exons and introns of the TPMT gene. Here we report the identification of a polymorphic locus within the promoter region of the gene. This polymorphism was detected by polymerase chain reaction - single strand conformation polymorphism analysis of DNA samples from 54 unrelated European individuals. A total of five alleles with length variations were distinguished through the 5'-flanking region involved in the TPMT gene expression. Sequence analysis revealed that these variations were due to a variable number of tandem repeats (VNTR), ranging from four to eight repeats. Each repeat consists of 17 or 18 bp units and contains putative binding sites for transcription factors. The most frequent alleles harbour four or five tandem repeats, a heterozygosity rate of 0.44 was calculated, and a stable Mendelian inheritance of alleles was demonstrated. Analysis of the effect of each VNTR allele on promoter activity of a reporter gene was further performed in various cell lines by transient transfection assay. A modulatory effect of VNTR alleles was observed in vitro, but the repeat polymorphism did not display a significative role in TPMT gene regulation in vivo. Further studies need to be carried out to support the hypothesis that VNTR may contribute to the large interindividual variations of TPMT activity. PMID- 10376767 TI - Functional analysis of the human D5 dopamine receptor missense and nonsense variants: differences in dopamine binding affinities. AB - The functional analysis of expressed human gene variants is important in the study of genetic susceptibility to diseases, pharmacogenetic traits and for the investigation of the human genetic diversity at the molecular level. We have performed the analysis of sequence polymorphisms in the human D5 dopamine receptor gene (DRD5) predicting missense and nonsense amino acid changes in the receptor protein. The amino acid substitutions in the human D5 dopamine receptor are: Leu88 to Phe in the putative second transmembrane domain, Ala269 to Val in the third intracellular and Pro330 to Gln in the third extracellular loops, Asn351 to Asp in the seventh transmembrane and Ser453 to Cys in the C-terminal domains and Cys335 to Stop in the third extracellular loop. The two amino acid substitutions in the transmembrane domains had an effect on agonist binding to the human D5 dopamine receptor. Asn351 to Asp resulted in an approximately 10 fold decrease in dopamine and threefold decrease in R(+)-SKF-38393 binding affinities. Leu88 to Phe resulted in a small increase in dopamine binding affinity. Antagonist binding affinities were mostly unaffected by the amino acid substitutions with the exception of Leu88 to Phe, which showed small reductions in binding affinities of SCH-23390 and risperidone. The existence of functionally different variants of the human dopamine receptors might have phenotypic consequences given their importance in central nervous system physiology and pharmacology. PMID- 10376768 TI - Characterization and substrate specificity of UGT2B4 (E458): a UDP glucuronosyltransferase encoded by a polymorphic gene. AB - Variations in glucuronidation activities among different individuals have been reported; however, genetic polymorphisms in the genes encoding phase II drug metabolizing UDP-glucuronosyltransferases have not been studied extensively. A novel UGT2B cDNA clone UGT2B4(E458) was isolated from human prostate and LNCaP cell cDNA libraries. The cDNA encoding UGT2B4(E458) is 2097 bp in length and has an open reading frame of 1584 nucleotides encoding a protein of 528 amino acids. Characterization of the UGT2B4(E458) cDNA revealed nucleotide differences with the previously published UGT2B4 and UGT2B11 cDNAs. These variations in the UGT2B4 sequence lead to an amino acid change from aspartic acid to glutamic acid at position 458. In the previous UGT2B11 cDNA (which has subsequently been renamed UGT2B4 (L109,396, D458)), leucine residues are found at positions 109 and 396, whereas phenylalanines are present at these positions in the UGT2B4(D458) and UGT2B4(E458) enzymes. Analysing the genomic DNA of 26 unrelated Caucasian individuals demonstrated the presence of variant alleles encoding UGT2B4(D458) and UGT2B4(E458). Stable expression of UGT2B4(E458) cDNA in HK293 cells demonstrates the presence of a 52 kDa protein, which is in agreement with other characterized (UGT2B proteins. UGT2B4(E458) conjugates hyodeoxycholic acid (HDCA) as well as 4-hydroxyestrone (4-OH-E1), androstane-3alpha,17beta-diol (3alpha diol) and androsterone (ADT). Specific reverse transcriptase-polymerase chain reaction analysis revealed expression of UGT2B4(D458) and UGT2B4(E458) transcripts in a wide range of extrahepatic tissues, including the liver, kidney, testis, mammary gland, prostate, placenta, adipose, adrenal, skin and lung. Our results suggest that UGT2B4(E458) and UGT2B(E458) are two widely expressed isoenzymes, and that polymorphism in the UGT2B4 gene might be responsible for differences in UGT2B4 enzymatic properties. PMID- 10376769 TI - Comparative evolutionary pharmacogenetics of CYP2D6 in Ngawbe and Embera Amerindians of Panama and Colombia: role of selection versus drift in world populations. AB - The development of CYP2D6 has been attributed to the need of earth-dwelling animals to detoxify toxic xenobiotics (phytoalexins) present in plants. This hypothesis has been extrapolated to humans, but is yet unconfirmed. Therefore, we studied two Amerindian populations as the best available model to test the effect of selection through diet on human CYP2D6 evolution. The frequency of sparteine poor metabolizers in Ngawbe was 4.4% (n = 344), while the frequency in Embera was 2.2% (n = 153). Among Ngawbe and Embera, CYP2D6*4 (allelic frequencies for each tribe, respectively: 0.171; 0.14), CYP2D6*6 (0.005; 0.011) and CYP2D6*10 (0.175; 0.069) were detected, while CYP2D6*3, CYP2D6*5, CYP2D6*9 and CYP2D6*16 were absent. All poor metabolizers possessed either CYP2D6*4 or CYP2D6*6 and there were no disagreements between genotypic and phenotypic data. The total frequency of mutant alleles showed no difference among Amerindians or when compared to Caucasians. It was higher than in Chinese, since the frequency of CYP2D6*4 was higher in Amerindians. XbaI restriction fragment length polymorphisms haplotypes were very homogeneous in Amerindians, because the only fragment that hybridized with the CYP2D6 cDNA probe was the 29 kb (not 42/44 kb or 11.5/13 kb). This indicated no gene cluster recombinations that generate insertions or deletions. We propose that in earlier hominids and humans, CYP2D6 had increasingly become a vestigial characteristic unconstrained by dietary stressors, as a result of cultural survival strategies. Human CYP2D6 evolution was preferentially affected by random genetic drift, and not by adaptive or purifying selection. PMID- 10376770 TI - An inhibitory monoclonal antibody to human cytochrome P450 2A6 defines its role in the metabolism of coumarin, 7-ethoxycoumarin and 4-nitroanisole in human liver. AB - Cytochrome P450 (CYP) 2A6 is an important enzyme catalysing the metabolism of many drugs, procarcinogens and promutagens. Its role in human liver metabolism of coumarin, 4-nitroanisole, 4-nitrophenol and 7-ethoxycoumarin was analysed with an inhibitory monoclonal antibody (MAb) to CYP2A6. MAbs were derived from a panel of 16 hybridomas which yielded positive enzyme-linked immunosorbent assay (ELISA) results or immunoblots against CYP2A6. The hybridomas were selected from more than 500 clones generated by the fusion of myeloma cells with spleen cells of mice immunized with purified baculovirus-expressed human CYP2A6. The MAbs obtained from four of the 16 hybridomas exhibited strong inhibitory activity to CYP2A6-catalysed phenanthrene metabolism. MAb 151-45-4 was positive and highly specific to CYP2A6 as determined by ELISA and immunoblot, and showed no cross reactivity with recombinant human CYP 1A1, 1A2, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, 3A4 and 3A5, as tested with ELISA and immunoblot analyses. MAb 151-45-4 specifically inhibited CYP2A6-catalysed metabolism of phenanthrene, 4 nitroanisole, 4-nitrophenol, coumarin and 7-ethoxycoumarin each by 94-99% and did not inhibit their metabolism catalysed by 10 other human CYPs. The potent inhibitory effect of MAb 151-45-4 was used to define the contribution of human CYP2A6 to the metabolism of coumarin, 4-nitroanisole and 7-ethoxycoumarin in seven human liver microsome samples. Coumarin metabolism in all of the seven samples was inhibited by greater than 94% by MAb 151-45-4 which indicates that essentially all microsome mediated coumarin metabolism in human liver is catalysed only by CYP2A6. Inhibition of 4-nitroanisole and 7-ethoxycoumarin metabolism by anti 2A6 MAb ranged from 22-65% and 8-24%, respectively. The degree of inhibition defines the contribution of CYP2A6 activity to the 4-nitroanisole and 7-ethoxycoumarin metabolism in human liver and the range reflects the variability among samples. The inhibitory antibody to CYP2E1 was used to determine its role in 4-nitroanisole and 7-ethoxycoumarin metabolism in seven human liver samples. The addition of both MAbs to CYP2A6 and 2E1 to the microsome samples defined combinatorially the relative role of CYP2A6 and 2E1 in the metabolism of 4-nitroanisole and 7-ethoxycoumarin. PMID- 10376772 TI - The hereditary transmission of the glutathione transferase hGSTT1-1 conjugator phenotype in a large family. AB - The polymorphism of human glutathione transferase hGSTT1-1 is expressed in three phenotypes. Experimentally, individuals can be classified as non-conjugators, low conjugators and 'high' conjugators depending on the enzyme activity in blood towards methylene chloride using a gas chromatographic assay. Non-conjugators do not have a functional hGSTT1 gene; however, little is known about the molecular basis of the three conjugator phenotypes. The higher hGSTT1-1 activity in high conjugators may be the result of enzyme induction or be genetically determined. Twenty-nine members of a large family, including three generations were phenotyped and genotyped with respect to hGSTT1-1. The hGSTT1-1 enzyme activity of high conjugators was twice as high as that of low conjugators. The distribution of hGSTT1-1 phenotypes strongly indicates a Mendelian intermediary inheritance, in which a gene-dosage effect results in a doubled enzyme expression in the presence of two functional alleles. The Mendelian intermediary inheritance is further supported by the findings of a semiquantitative polymerase chain reaction method designed to distinguish the three genotypes of hGSTT1 for rapid screening of large study groups. PMID- 10376771 TI - Genetic variation in the metabolism of coumarin in mouse liver. AB - The metabolism of 50 microM [3-14C] coumarin to polar products separated by high performance liquid chromatography (HPLC) and covalently bound metabolites in liver microsomes was compared in a series of inbred strains of mice. Coumarin metabolism to total polar products was higher in female than male mice. In all strains, the coumarin 3,4-epoxidation pathway was the major route of metabolism with o-hydroxyphenylacetaldehyde (o-HPA) as the major metabolite. However, in females, there was a major strain difference in the degree of metabolism to coumarin 7-hydroxylase with DBA/2 and 129 having high 7-hydroxycoumarin formation, CBA/Ca having intermediate levels and the other strains low levels. The differences between the strains was much less pronounced in the male mice. There was also evidence for strain variation in metabolism in the quantities of a number of other coumarin metabolites as detected by HPLC analysis of incubate extracts. However, this variation was of a quantitative nature and relatively small. The metabolism of B6C3F1 hybrid mice, in which coumarin had been identified as carcinogenic in a long-term cancer bioassay, was qualitatively similar to that of the other genotypes. The DBA/2 mouse has been suggested as a model for the metabolism of coumarin in humans. The pattern of metabolism found in this strain is different from most other strains. However, the pattern found for all the mouse strains, including DBA/2, differed appreciably from the profiles for other species including humans in the extent of 7-hydroxylation. PMID- 10376773 TI - Thiopurine methyltransferase pharmacogenetics: alternative molecular diagnosis and preliminary data from Northern Portugal. AB - The thiopurine methyltransferase (TPMT) genetic polymorphism has been shown to have a highly significant clinical impact, namely in the therapeutic efficiency of thiopurine drugs used in the treatment of a wide range of diseases. Available diagnostic methods, although reproducible and sensitive, are relatively laborious. Thus population studies are still very scarce. In this work we describe a new polymerase chain reaction-single strand confirmational analysis based protocol for TPMT specific detection which introduces a substantial technical simplification avoiding the use of restriction enzyme treatment after polymerase chain reaction amplification. Additionally, the use of this protocol allows the simultaneous detection of a T474 to C substitution, a frequent silent mutation in the North Portuguese population (TPMT*1S = 0.215). In a sample of 310 unrelated Northern Portuguese individuals, 15 were found to be heterozygous for the TPMT*3A allele (defined by the presence of two transitions, G460 to A and A719 to G) which is associated with TPMT enzymatic deficiency; the corresponding gene frequency estimate was 0.024. We also attempted to evaluate the relationship between the molecular TPMT genotype and the reaction to treatments involving thiopurine drugs by analysing a sample of 24 children submitted to curative therapy of acute lymphoblastic leukaemia. Four of them were shown to be heterozygous for the TPMT*3A allele. An examination of their clinical histories showed that all four patients exhibited signs of severe hepatic toxicity during treatment. PMID- 10376774 TI - Cytochrome P450 3A proteins are expressed in B lymphocytes but not in T lymphocytes. PMID- 10376775 TI - Dietary-fat intake in the US population. AB - OBJECTIVE: This article looks at the food group choices by individuals grouped based on fat intake, saturated fat intake, and use of lowfat foods. METHODS: Food consumption data from USDA's National Food Consumption Surveys (NFCS) and the Continuing Survey of Food Intakes by Individuals (CSFII) were used to look at changes in the mean energy, percent calories from fat and saturated-fat and total fat intakes over time. USDA's 1995 CSFII data were used to evaluate the diets of individuals grouped based on percent calories from fat and use of low-fat foods. Individuals six to 50 years old who had complete food intake records were included and five age-gender classifications were used. RESULTS: The percent of calories from total fat and saturated fat have steadily declined over the last 30 years, and the amount of fat in the diet has increased from 1989 to 1995. Those whose diets met the Dietary Guidelines Recommendations for fat and saturated fat had lower fat intakes. Except for adult males, those with low-fat diets had higher intakes of total-food amount. Also, lower saturated-fat intakes were associated with lower energy intakes. In general, high-fruit-and-grain-products consumption were seen in groups with low-fat intake. For those who included low fat foods in their diets and also had low-fat intakes, rice and pasta were the major foods of choice for calories. Fried potatoes were one of the main sources of calories for high-fat groups. CONCLUSION: The study showed individuals whose diets included low-fat foods are more likely to have a diet that meets the dietary guidelines recommendations for fat and saturated fat. PMID- 10376776 TI - Vitamin A and zinc supplementation of preschool children. AB - OBJECTIVE: To determine whether supplementation of vitamin A and/or zinc (Zn) improved serum levels of these nutrients and/or height and weight gains in preschool children, 22 to 66 months, living in Belize, Central America. METHODS: Subjects received either Zn, vitamin A, Zn and vitamin A or a placebo, (70 mg Zn and/or 3030 RE vitamin A, once per week) for 6 months in a 2x2 factorial design. Forty-three children, from a population of 104 prescreened, completed the study; they were selected, prior to treatment, for low/marginal serum concentrations of these micronutrients. RESULTS: Serum Zn levels were greater (16%, p<0.001) for those who received Zn. In contrast, after vitamin A treatment there were no differences in serum vitamin A among groups. Although increases in height (+4.4 cm, p<0.001) and weight (+0.79 kg, p<0.001), compared with baseline values, were numerically greatest for children who received both supplements, only the vitamin A supplementation effect was significant, resulting in increased height (+1.4 cm, p<0.002) and greater weight gain (+0.15 kg, p<0.03) compared to those receiving no vitamin A. Vitamin A supplementation alone significantly increased (p<0.001) hemoglobin concentration. CONCLUSION: The results suggest that the preschool children in this study, prescreened for low/marginal serum concentrations from a larger population prior to treatment, were enduring inadequate vitamin A and, to a lesser degree, Zn nutriture. Height and weight gain were significantly increased in the subjects who received a single weekly supplement 3030 RE of vitamin A. PMID- 10376777 TI - Weight satisfaction and dieting practices among college males in Taiwan. AB - OBJECTIVE: To obtain baseline data regarding body mass index (BMI), weight satisfaction and weight perception in male college students. Approaches used to lose weight and characteristics of dieters were also investigated. METHODS: Written questionnaires and height and weight measurements were used to collect data. Nine-hundred thirty male college students selected by multiple stage sampling among colleges in Taiwan participated in this study. Simple frequency and ANOVA were used to analyze data. Post hoc analyses were performed with the LSD test when the F ratio for the ANOVA was significant at p<0.05. RESULTS: The mean BMI for all subjects was 21.7. Males with a BMI < or =22.5 are considered thin by Taiwanese and world standards. Yet 34% of these males were attempting to lose weight and 14% percent perceived themselves as either overweight or obese. Exercise was the main approach to losing weight for subjects in all weight categories. Subjects in the dissatisfied/dieting group tended to measure body weight more frequently (p<0.05), spent more time exercising and reading nutrition information (p<0.05) and skipped breakfast and lunch more frequently (p<0.05) than non-dieters. Male dieters ate fewer vegetables and less meat than non dieters. CONCLUSIONS: Male college students had frequent misconceptions and dissatisfaction with their body weight. Behavioral characteristics among those dissatisfied with their weight indicate they are at risk for developing eating disorders. PMID- 10376778 TI - Effect of pistachio nuts on serum lipid levels in patients with moderate hypercholesterolemia. AB - BACKGROUND: Elevated serum cholesterol levels play an important role in the development of coronary artery disease. Previous studies have suggested that nut consumption benefits lipid profile. Pistachio nuts are widely available, inexpensive and frequently consumed by the general population. OBJECTIVE: To determine whether substituting 20% of the daily caloric intake in the form of pistachio nuts will improve the lipid profiles of humans with primary, moderate hypercholesterolemia. DESIGN: Controlled, randomized crossover design. SETTING: Outpatient dietary modification, counseling and blood analysis. PATIENTS: Ten patients with moderate hypercholesterolemia. INTERVENTION: Three weeks of dietary modification with 20% caloric intake from pistachio nuts. MEASUREMENTS: Body weight, blood pressure, total cholesterol, LDL, HDL, and triglycerides were monitored. Lipid profiles were analyzed prior to, during and after dietary modification. RESULTS: After three weeks, there was a decrease in total cholesterol (p<0.04), an increase in HDL (p<0.09), a decrease in the total cholesterol/HDL ratio (p<0.01) and a decrease in the LDL/HDL ratio (p<0.02). Triglycerides and LDL levels decreased, but not significantly. Body weight and blood pressure remained constant throughout the study. CONCLUSIONS: Results suggest that eating pistachio nuts instead of other dietary fat calories can improve lipid profiles, thereby decreasing coronary risk. Further studies will be required to confirm these results and to determine the mechanism of this effect. PMID- 10376779 TI - Comprehensive assessment of the components of energy expenditure in infants using a new infant respiratory chamber. AB - BACKGROUND: Current methods for energy expenditure (EE) measurements in term infants do not include simultaneous measurements of basal and sleeping metabolic rates (BMR and SMR) or a measure of physical activity (PA). Furthermore, prediction equations for calculating EE are not appropriate for use in infants with metabolic disorders. OBJECTIVE: To develop and utilize a new infant respiratory chamber for simultaneous measurements of EE (kJ/d), preprandial BMR (kJ/d), SMR (kJ/d) and an index of PA (oscillations/min/kg body weight) in infants with a variety of metabolic disorders, for up to four hours in a hospital setting, while allowing parental interaction in a comfortable environment. METHODS: We obtained simultaneous measurements of EE, BMR, SMR and PA in 21 infants (66+/-73 days of age, 4.5+/-1.7 kg body weight, 55+/-8 cm in length and 16+/-7% body fat) using our new infant respiratory chamber. Six of these infants were healthy, seven had thyroid dysfunction, five were HIV-exposed, one had AIDS, one had intrauterine and postnatal growth retardation and one was a hypothermic preterm infant. Energy expenditure, BMR and SMR were extrapolated for 24 hours. Body composition was estimated by skin-fold thickness, using age-appropriate formulae. Basal metabolic rate obtained with the infant respiratory chamber was compared to BMR that was calculated using the appropriate World Health Organization (WHO) equations. RESULTS: In all infants both extrapolated 24-hour EE and BMR correlated with fat-free mass (r = 0.89, p<0.01 and r = 0.88, p<0.01 respectively). Twenty-four hour EE also correlated with PA (r = 0.52, p<0.05). The HIV-exposed infants had higher BMR (p<0.05) than that calculated by the appropriate WHO equation. We found that the caloric requirements for the infant with growth retardation were underestimated based on the infant's weight and age. CONCLUSIONS: The infant respiratory chamber can measure all of the main components of EE. Some of the results obtained differed significantly from those obtained by the WHO equations; therefore, the new infant respiratory chamber is necessary for estimating EE in infants with metabolic and growth disorders. PMID- 10376781 TI - Patterns in child and adolescent consumption of fruit and vegetables: effects of gender and ethnicity across four sites. AB - OBJECTIVES: Few studies have examined the association of gender and ethnicity with fruit and vegetable consumption. We examined these associations using baseline data from four school-based sites funded under the National Cancer Institute's 5 A Day for Better Health Program. METHODS: Diet was measured using 24-hour recalls at three sites and seven-day food records at one site. Demographics were obtained via self-report or school records. Regression analyses for clustered data were employed with fruit and vegetables combined and fruit and vegetables separately. RESULTS: Girls ate more fruit, more vegetables and more fruit and vegetables combined than boys at the Georgia site. Ethnicity was significant in two sites: In Georgia, African-Americans ate more fruit and more fruit and vegetables combined than European-Americans; in Minnesota, Asian American/Pacific Islanders and African-Americans ate more fruit than European Americans, and European-Americans and African-Americans ate more vegetables than Asian-Americans. No significant effects were found at the Alabama or Louisiana sites. CONCLUSIONS: Ethnicity was related to fruit and vegetable consumption in Georgia and Minnesota. Consistent with prior studies, gender was related to fruit and vegetable consumption, with girls consuming more servings than boys; however, this was observed at one site only, Georgia. Consumption levels were similar to national estimates for children and varied by region. Further studies are needed using a single methodology to facilitate regional comparisons. PMID- 10376780 TI - Day-to-day consistency in amount and source of carbohydrate intake associated with improved blood glucose control in type 1 diabetes. AB - OBJECTIVE: To determine if a relationship exists between blood glucose control and variability in nutrient intake from day-to-day in subjects with type 1 diabetes. METHODS: Two three-day diet records and one measurement of glycated hemoglobin (HbA1c) were obtained from 272 subjects with type 1 diabetes treated with a mixture of regular and NPH insulins before breakfast and supper and using a standardized algorithm to adjust insulin dose according to the results of self monitoring of blood glucose two to four times daily. Day-to-day variation in nutrient intake was expressed as the coefficient of variation (CV = SDx100/mean). RESULTS: Nutrient intakes in the study population (mean +/- SD) were energy 8.35+/-2.43 MJ, fat 81+/-30 g, protein 94+/-28 g, carbohydrate 227+/-68 g, starch 126+/-38 g and dietary fiber 20+/-6 g with diet glycemic index being 84.2+/-7.4. Neither energy, nutrient intakes nor insulin dose was significantly related to HbA1c. Day-to-day variation of carbohydrate (p = 0.0097) and starch (p = 0.0016) intakes and diet glycemic index (p = 0.033) was positively related to HbA1c, and the associations remained significant when adjusted for age, sex, duration of diabetes and BMI. Day-to-day variation in energy, protein or fat intakes was not related to HbA1c. CONCLUSIONS: Consistency in the amount and source of carbohydrate intake from day-to-day is associated with improved blood glucose control in people with type 1 diabetes, a result which supports continued educational efforts to achieve adherence to a diabetes diet plan. This conclusion may not apply to people on intensified insulin therapy who adjust their insulin dose based on their actual carbohydrate intake at each meal. PMID- 10376782 TI - Variability in the trans fatty acid content of foods within a food category: implications for estimation of dietary trans fatty acid intakes. AB - OBJECTIVE: Currently, the published information on trans fatty acid composition of foods is incomplete and of questionable accuracy. Detailed fatty acid analysis of over 200 foods was undertaken for the purpose of determining the variability in trans fatty acid content among foods within a product category, and the significance of this variability to the estimation of trans fatty acids intakes from analysis of dietary intake data. METHODS: The analysis of food fatty acids used gas-liquid chromatography with 100 m capillary columns and standardized methodologies for food sampling, fat extraction, separation and quantification of trans fatty acid isomers. For the purposes of this report, trans refers to all non-naturally occurring isomers including trans, cis-trans, geometric and positional isomers. RESULTS: The results show that the amount of trans fatty acids varies considerably among foods within a category, reflecting differences in the fats and oils used in the manufacturing or preparation process. For example, the range of trans fatty acids in 17 brands of crackers was 23 to 51% total fatty acids, representing differences of from 1 to 13 g trans fatty acids per 100 g cracker. The large errors that may arise in estimates of the trans fatty acid intake of an individual are illustrated by analyses of the potential trans fatty acid intake in a sample diet, for each food as calculated using the minimum and maximum values for trans fatty acids within a given category. The results of these analyses show estimates of trans fatty acid intake from a low of 1.4 to 25.4 g a day for the same diet. CONCLUSION: This study shows that the wide variability in trans fatty acid content of different foods may result in large errors in the estimation of trans fatty acid intake of individuals and, potentially, groups. PMID- 10376783 TI - Zinc and IGF-I concentrations in pregnant women with anemia before and after supplementation with iron and/or zinc. AB - OBJECTIVE: The objective of our study was to investigate zinc (Zn) status and effects of Zn supplementation in relation to insulin-like growth factor-I (IGF-I) and iron deficiency anemia in pregnant women. The role of Zn and IGF-I in hematologic abnormalities has remained unclear. METHODS: Thirty-eight Japanese women, when examined at the second trimester of pregnancy, had hemoglobin concentrations below 11.0 g/dL and 32 of 38 had normocytic erythrocytes. These 38 women were divided into three groups, and we compared the hematological status and serum IGF-I levels before and after iron (Group A) or Zn (Group B) or iron plus Zn (Group C) supplementation. RESULTS: The concentrations of hemoglobin (Hb) did not change in groups A and B. In group C, Hb levels were significantly increased from 10.3+/-0.3 to 11.0+/-0.6 g/dL. Furthermore, numbers of RBC and reticulocytes also increased significantly. Concentrations of iron, IGF-I and total iron binding capacity (TIBC) were increased, and concentrations of erythropoietin were decreased, but not statistically. There were significant positive correlations between increases in IGF-I and increases in Hb and RBC in the Zn administered groups. CONCLUSION: Zn status to some extent can account for hematological abnormalities in pregnant women. Zn derived IGF-I has a role in the regulation of hematopoiesis in pregnant women. PMID- 10376784 TI - Body fat percent by bioelectrical impedance analysis and risk of coronary artery disease among urban men with low rates of obesity: the Indian paradox. AB - OBJECTIVE: To determine the association between body fat percent and prevalence of coronary artery disease (CAD) and coronary risk factors in subjects with low rates of obesity. SUBJECTS AND METHODS: We randomly selected 850 men, aged 25 to 64 years. The survey methods were questionnaire and bioelectrical impedance analysis for body composition. Subjects were divided into high-fat (n = 357), over-fat (n = 230), normal-fat (n = 200) and under-fat (n = 63) based on criteria of body-fat percent analysis. RESULTS: The prevalence of CAD and the coronary risk factors hypercholesterolemia, hypertension, diabetes, mellitus and sedentary lifestyle were significantly associated with high and moderate body fat percent despite low body-mass index (23.6+/-4.1 kg/m2). Mean total cholesterol, triglycerides and blood pressure were significantly associated with high and moderate body fat percent. The prevalence of smoking was weakly but inversely associated with high body-fat percent. Mean HDL cholesterol was positively associated with high body-fat percent. Body mass index was positively associated with high body-fat percent. CONCLUSIONS: High and moderate body-fat-percent subjects were associated with high prevalence of CAD and the coronary-risk factors hypertension, diabetes mellitus, higher body-mass index and sedentary lifestyle. PMID- 10376785 TI - Nutritional parameters and short term outcome in arthroplasty. AB - OBJECTIVE: Advances in surgical techniques and management of arthroplasty patients have contributed to a significant reduction in surgical complication rates. Preoperative nutritional status has a significant impact on surgical outcome. Studies have reported improved outcomes in burn and hip fracture patients receiving nutritional supplementation during their recoveries. Our objective was to assess the effects of preoperative nutritional status on the incidence of complications, resource consumption, and length of stay of patients undergoing hip and knee replacement surgery. METHODS: One hundred and nineteen patients were evaluated. Standard preoperative laboratory tests were performed on all patients. Medical severity of illness was assessed on all patients using the Charlson Comorbidity Index. Anesthesia and surgical time was recorded. Short term outcome was assessed utilizing hospital charges as a measure of resource consumption, length of stay (LOS), in-hospital consults and the presence and number of complications during hospitalization. Non-parametric Kruskall Wallis and chi-square statistical analyses were performed. A p value <.05 was considered significant. RESULTS: Mean age was 64.6 years +/-15.62. 52.9% had osteoarthritis (OA), 4.2% had rheumatoid arthritis (RA), 5.9% had osteonecrosis (ON), 9.2% had a hip fracture and 28% had a failed total knee arthroplasty (TKA) or total hip arthroplasty (THA). Mean albumin and total lymphocyte count (TLC) were 38.5 g/L +/-4.78 SD and 1884 cells/microL +/-762 SD, respectively. Patients with albumin levels less than 34 g/L had 32.7% higher charges ($50,108+/-8203 SE vs. $33,720+/ 1128 SE, p<.006), higher medical severity of illness (p = .03) and longer LOS (8.6+/-1.7 SE vs. 5.2+/-.356 SE days, p<.001). Patients with TLC less than 1200 cells/microL had higher charges ($32,544+/-1050 SE vs. $42,098+/-3122 SE, p = .004), longer LOS (5.7+/-.531 vs. 5.4 days +/-.368, p = .004) and anesthesia (242.85+/-17.55 SE vs. 198.6 min. +/-6.06 SE, p = .02) and surgical times (177.14 min. +/-17.57 SE vs. 120.21 min. +/-6.22 SE, p = .002) when compared with patients with TLC higher than 1200 cells/microL. These findings were still significant when adjusted for medical severity of illness and age. CONCLUSIONS: Our data demonstrate that preoperative nutritional status is an excellent predictor of short term outcome. Serum albumin and TLC correlate with resource consumption, length of stay and operative time in patients undergoing joint replacement surgery. These parameters may be improved with nutritional supplementation prior to surgery. PMID- 10376786 TI - Infant feeding practices of Anglo American and Asian Indian American mothers. AB - OBJECTIVE: To compare infant feeding practices of Anglo-American (AA) (n = 25) and Asian-Indian American (AIA) mothers (n = 25) residing in the southeastern United States. METHODS: Feeding practices (breast-feeding, formula-feeding, introduction of solid foods) were assessed at infant ages one, three, six, nine and twelve months for a total of 250 interviews conducted in the home. Mothers' sources of information about infant feeding practices and dietary intakes of their infants were collected (24-hour recalls). RESULTS: Compared to their AIA counterparts, AA mothers breast-fed for significantly longer durations and introduced formula and solid foods into the infants' diet at a later age (p<0.05). Throughout the first year, AA mothers relied primarily upon health professionals for infant feeding information compared to AIA mothers, who sought information primarily from the family network during the first six months and relied more on health professionals during the second six months of the infant's life. Throughout the first twelve months, infants of both groups exceeded 100% of the RDA for energy, protein, calcium, iron, vitamin A, and vitamin C. CONCLUSION: Health professionals, including nutrition educators, should educate AIA mothers about and encourage AA mothers to follow current feeding recommendations and guidelines about breast-feeding, formula-feeding and introducing solid foods. PMID- 10376787 TI - CD-ROMs for nutrition education. PMID- 10376788 TI - Tryptamine-mediated stabilization of tryptophanyl-tRNA synthetase in human cervical carcinoma cell line. AB - Tryptamine is an endogenous neuroactive metabolite of tryptophan. Interpretation of the function of this bioamine, however, is restricted to manipulation with tryptamine synthetic pathways. Meanwhile, tryptamine is a potent inhibitor of protein biosynthesis, via the competitive inhibition of tryptophanyl-tRNA synthetase (TrpRS). The influence of the persistent tryptamine inhibition on the half-life and cellular content of TrpRS was examined by chase labeling of HeLa cells and the tryptamine-resistant subline with [35S]methionine. The results indicate that long-term tryptamine treatment of HeLa cells led to a significant increase in the half-life of TrpRS while the content, in vivo phosphorylation and gene dose of TrpRS were unchanged. These findings suggest that survival of drug resistant cells may not be due to TrpRS gene amplification, but to stabilization of TrpRS. It was shown that tryptamine is an effective inhibitor of HeLa cell growth. In contrast to the well-characterized antineoplastic compounds, conferring a many hundred-fold elevated drug resistance to tumor cells, resistance to tryptamine at very low levels was difficult to achieve, i.e. the 2 fold resistant subline was selected after 19 months of treatment of HeLa cells with gradually increasing concentrations of tryptamine. The tryptamine-resistant HeLa subline exhibited a slower growth rate than the original HeLa line when similar concentrations of both cell populations were seeded on the plates. A low tryptamine resistance and a lack of TrpRS gene amplification were observed in two tryptamine-resistant HeLa sublines and three Chinese hamster sublines. The role of TrpRS in oncogenesis and the perspective for tryptamine as a potential anti cancer drug are discussed. PMID- 10376789 TI - Alteration of protein kinase C isoform-specific expression during rat hepatocarcinogenesis after exposure to the peroxisome proliferator WY-14,643. AB - The role of protein kinase C (PKC) isoforms in mediating peroxisome proliferator chemical- (PPC) induced hepatocarcinogenesis was examined. After an acute gavage exposure to WY-14,643 (WY) membrane-bound PKCdelta and cytosolic PKCbeta decreased, whereas the expression of the other isoforms was not altered. After a 13-week chronic exposure, membrane-bound PKCbeta, delta and zeta levels decreased. In WY-induced hepatocellular adenomas, PKCalpha was increased, and PKCbeta was further decreased in membrane fractions. These results, taken together with previous studies, indicate that alterations in PKCalpha, beta and delta isoforms, which regulate mitogenesis, could play important roles in perpetuating the high cell proliferative rate in PPC-induced hepatocellular adenomas. PMID- 10376790 TI - The inhibitory action of BOF-A2, a 5-fluorouracil derivative, on squamous cell carcinoma. AB - Inactivation of antineoplastics is a serious problem in cancer therapy, and prevention of the inactivation is a surpassing strategy for enhancement of their therapeutic effects. BOF-A2, which contains both EM-FU, a masked form of 5 fluorouracil (5-FU), and CNDP, an inhibitor of 5-FU degradation, was developed with this aim. We compared the antitumor effects of BOF-A2 and 5-FU in human squamous cell carcinoma cells transplanted to nu/nu mice. Each drug (0.9-7.0 mg/kg of 5-FU and 3.8-30 mg/kg of BOF-A2) was orally administered every day to 5 mice in each dosage group for 4 weeks. Although the maximal tumor growth inhibition by 5-FU (3.5 mg/kg per day) was about 50% of the control value, 15 mg/kg per day of BOF-A2, which is equimolar to 3.5 mg/kg per day of 5-FU, almost completely inhibited the tumor growth. The flow-cytometric analysis revealed that BOF-A2 (15 mg/kg per day) induced more prominent S-phase-accumulation (63 +/- 6%) of tumor cells than did 3.5 mg/kg per day of 5-FU (43 +/- 18%), and immunohistochemical stainings indicated that the decrease of proliferating cell nuclear antigen expression was more prominent in tumor cells in the BOF-A2 treated mice than in the 5-FU-treated mice. Correspondingly, the DNA synthesis was markedly suppressed in tumor cells obtained from BOF-A2-treated mice. Compared with 5-FU, BOF-A2 more strongly induced apoptosis; apoptotic cells detected by nick-end labeling techniques were about 20% of the tumor cells in 5 FU (3.5 mg/kg per day)-treated mice, and nearly 50% in BOF-A2 (15 mg/kg per day) administered mice. The expressions of involucrin, cytokeratin 10 and protein kinase C eta, which are associated with squamous cell differentiation, were not increased by BOF-A2 or 5-FU, although the expression of transglutaminase was slightly augmented by both drugs. These results indicate that compared with 5-FU, BOF-A2 more strongly suppresses the growth of squamous cell carcinoma by inhibiting DNA synthesis and inducing apoptosis but not cell differentiation. PMID- 10376791 TI - Prognostic significance of DNA ploidy pattern in osteosarcomas in association with chemotherapy. AB - In this study, we analysed the DNA ploidy of osteosarcomas at biopsy and attempted to clarify the relationship between DNA ploidy pattern and prognosis. Thirty patients with non-metastatic osteosarcoma of an extremity were studied. All underwent intensive chemotherapy with doxorubicin, cisplatin and methotrexate, in addition to wide tumor resection. DNA ploidy was detected by DNA cytofluorometry, using isolated and smeared cells of biopsied tumor tissue. Twelve tumors showed a diploid ploidy pattern and 18 showed a non-diploid pattern such as aneuploidy (15 tumors) and euploid-polyploidy (3 tumors). The event-free survival rate at 9 years was 63.5% in non-diploid osteosarcoma patients and 13.3% in diploid osteosarcoma patients. There was a statistically significant difference between the two groups (P = 0.0278). These results lead us to conclude that a non-diploid osteosarcoma may be more sensitive to chemotherapy than a diploid tumor. PMID- 10376792 TI - Tumor growth inhibition and regression induced by photothermal vascular targeting and angiogenesis inhibitor retinoic acid. AB - The effect of photothermal vascular targeting, alone and in combination with antiangiogenic therapy, was evaluated using tumors produced in mice by transplantation of KB cells. Tumor growth inhibition and regression followed vascular damage produced by pulsed dye laser (PDL) radiation. Administration of the antiangiogenic agent all-trans-retinoic acid (RA) was associated with smaller average tumor volumes in the presence and absence of PDL irradiation, but this effect was not statistically significant. The ability of PDL photothermal vascular targeting to cause regression of tumors without harming normal tissue may be a consequence of preferential damage to supplying vessels at the tumor periphery. PMID- 10376793 TI - Expression of hsp90 and cyclin D1 in human breast cancer. AB - The integral roles of heat shock proteins (hsps) in the cell cycle and in multistep processes leading to tumorigenesis have been implied. We examined the expression of hsp90alpha, hsp90beta and cyclin D1 in human breast cancer. Levels of mRNAs coding for hsp90alpha and cyclin D1 were significantly higher in cancer tissues than in non-cancer tissues. Moreover, there was a close relationship between the extent of the two mRNA levels, suggesting that increased expression of hsp90alpha, an isoform of the hsp90 family, is associated with the proliferation of human breast cancer. Hsp90beta was expressed in cancer cells, but not associated with cell proliferation. PMID- 10376794 TI - Lung cancer risk of the GSTM1 null genotype is enhanced in the presence of the GSTP1 mutated genotype in male Japanese smokers. AB - The potential interaction of GSTM1 and GSTP1 genotypes in pulmonary carcinogenesis was assessed in 382 male Japanese lung cancer patients (127 squamous cell carcinoma, 78 small cell carcinoma, 177 adenocarcinoma) and 257 controls. In smokers (358 cases, 184 controls) the GSTM1 null genotype was more distributed in individuals with at least one GSTP1 mutant allele compared to those without, in lung cancer patients (69.5% vs.53.2%) but not in controls (48.0% vs. 48.5%). No such relationship was detected in non-smokers (24 cases, 73 controls). The estimated relative risk of the GSTM1 null genotype for lung cancer was 2.58 (95%CI = 1.26-5.30) in smokers with the GSTP1 mutant allele while it was 1.17 (95%CI = 0.77-1.79) in those without, suggesting that mutated GSTM1 and GSTP1 genotypes interact to potentiate the risk of lung cancers in Japanese smokers. PMID- 10376795 TI - Reduced expression of protein tyrosine phosphatase gamma in lung and ovarian tumors. AB - Based on LOH studies protein tyrosine phosphatasegamma (PTPgamma) has been suggested as a candidate tumor suppressor gene involved in the oncogenesis of lung and renal cancers. In order to assess the involvement of PTPgamma in tumor development we developed a PTPgamma-specific monoclonal antibody (gammaTL1) (IgM isotype) by immunization with a synthetic peptide of 15 amino acids corresponding to the amino acid sequence nos. 1423-1438 just outside the phosphatase domain-II. In line with the fact that the antibody was raised to an intracellular domain of the PTPgamma molecule the antibody labeled the cell membrane of fixed cells but did not stain the outside of the cell membrane in the immunofluorescence assay. The Mab gammaTL1 recognized a full-length baculovirus recombinant PTPgamma protein of 185 kDa, in addition to putative cleavage products of 120 kDa, 114/110 kDa and 80 kDa, on Western blots of lysates of PTPgamma-gene transfected Sf9 insect cells but not of tumor cell lysates. Based on immunoperoxidase and immunofluorescence assays on cryostat sections, however, PTPgamma was expressed in more than 90% of both normal, human tissue samples and in the (non-) tumor cells of carcinoma samples. However, PTPgamma was not found in 28% of the overall lung tumor samples, i.e. in 50% of the lung adenocarcinoma samples, while the expression was weak and heterogeneous in 71% of squamous lung cell carcinomas. PTPgamma was not suppressed in the normal cells between the lung carcinoma cells. The presence of PTPgamma, assayed by immunofluorescence in lung tumor cell lines (H69, H128, H82, C3) was confirmed by RT-PCR assay. Interestingly, the 90% expression score of PTPgamma protein in normal ovarian tissue samples was reduced dramatically to 44 and 38% in both the non-tumorous and tumorous cells, respectively, in ovarian tumor samples. PTPgamma was absent in the HT29 human colon carcinoma cell line both by immunofluorescence and RT-PCR assay. In summary, we have developed a PTPgamma-specific monoclonal antibody, that demonstrated that the expression of PTPgamma is severely reduced (>50%) in lung tumors and ovarian tumors. PMID- 10376796 TI - Cytotoxic activity of pierisin, from the cabbage butterfly, Pieris rapae, in various human cancer cell lines. AB - Pierisin, a protein purified from pupae of the cabbage butterfly, Pieris rapae, exhibits cytotoxic effects against the human gastric cancer TMK-1 cell line, inducing apoptosis. The present study was performed to determine whether pierisin might exert a similar influence on nine other human cancer cell lines and human umbilical vein endothelial cells (HUVECs). Pierisin showed cytotoxic effects in all the human cells tested, with IC50 values ranging from 0.043 ng/ml to 150 ng/ml. Among the target cells, the cervical carcinoma cell line, HeLa, was the most sensitive to pierisin, showing a 1000-fold less IC50 value than that of HUVECs. While pierisin clearly induced apoptotic cell death in most cancer cell lines and HUVECs, the pathway appeared to be probably different from that involving anti-Fas, TNF-alpha and p53. Pierisin may thus be a promising new candidate for cancer therapy. PMID- 10376797 TI - Tumor-enhancement effect of a Mn3+ metalloporphyrin derivative (ATN-4T) in magnetic resonance imaging. AB - Magnetic resonance imaging is routinely used for tumor recognition in cancer diagnosis. The tumor image-enhancing characteristics of ATN-4T (THF-Mn-Asp), a Mn3+ metalloporphyrin derivative, were evaluated in rabbits. ATN-4T (10 mM) was diluted in gelatin to final concentrations ranging from 100 to 1 microM. Increasing concentrations of ATN-4T resulted in higher signal intensities. VX2 (squamous cell carcinoma) tumor-bearing rabbits were injected with 50 micromol/kg ATN-4T intravenously and T1 -weighted images were recorded continuously. Tumor images were compared with images of surrounding muscle tissue. T1-weighted images from ATN-4T-treated rabbits showed a marked enhancement of tumor contrast from 30 to 240 min postinjection. Microscopic examination revealed that carcinoma cells were scattered throughout the high contrast area of the tumor and were not seen in the surrounding muscles. ATN-4T appears useful for enhancing the intensity of tumors imaged by magnetic resonance. PMID- 10376798 TI - Contribution of myeloid and lymphoid host cells to the curative outcome of mouse sarcoma treatment by photodynamic therapy. AB - Selective depletion or inactivation of specific myeloid populations (neutrophils, macrophages) and lymphoid populations (helper T cells, cytolytic T cells) in EMT6 sarcoma-bearing mice was used to determine the contribution of each of these host immune cell types to the curative outcome of Photofrin-based photodynamic therapy (PDT). Immunodepletion of neutrophils and cytolytic T cells initiated immediately after PDT resulted in a marked reduction in PDT-mediated tumor cures. Significant reduction in the cures of EMT6 tumors was also achieved by immunodepletion of helper T cells and inactivation of macrophages by silica treatment. The initial tumor ablation by PDT was not affected by any of the above depletion treatments. These results provide direct evidence that the contribution of neutrophils, macrophages and T lymphocytes is essential for the maintenance of long-term control of PDT-treated tumors. PMID- 10376799 TI - The in vivo effect of the administration of resistance-modulating agents on rhodamine 123 distribution in mice thymus and lymph nodes. AB - P-glycoprotein (Pgp) has been widely associated with the multidrug resistance phenotype. Nevertheless, this protein has been detected in many normal tissues and cells, including liver, kidney, endothelial cells that constitute the hematological barrier of the brain and testes, and cells from the immune system. Many in vitro models have been used to study drugs that modulate Pgp activity and the multidrug resistance phenomenon. In the present work, we investigate the in vivo effects of resistance-modulating agents on lymphoid organs. Rhodamine 123 (Rho123), a well-known Pgp substrate, was administered to mice, and the fluorescence level in thymus and lymph node cells measured. The fluorescence level on these organs showed a dose-dependent response. Cyclosporin A (CSA), Verapamil (VP) and Trifluoperazine (TFP), three resistance-modulating agents, were administered to mice 1 h prior to 1 mg/kg Rho123 administration. Surprisingly, VP (10 mg/kg) and TFP (750 microg/kg) did not modulate Rho123 retention by thymus and lymph node cells. CSA (50 mg/kg) was the only drug that increased the fluorescence level in both organs. These results point out to the need of a wider study on the in vivo effects of resistance-modulating agents in different organs and systems. PMID- 10376800 TI - Simian virus 40 T antigen induces p53-independent apoptosis but does not suppress erbB2/neu gene expression in immortalized human epithelial cells. AB - Previously, we have shown that simian virus 40 (SV40) T antigen can directly cause apoptosis in immortalized human epithelial cells under normal growth conditions. In this study, we further characterized the mechanism of T-antigen mediated apoptosis involving p53 and whether T antigen can suppress erbB2/neu gene expression. Our results show the differential regulation of erbB2/neu gene expression in different cell clones in response to T antigen transgene, indicating that the regression of erbB2/neu gene by SV40 T is cell-type dependent. Our previous study reported T-antigen-induced apoptosis in p53 mutant cells; however, in this study we find increased levels of p53 protein in T antigen-containing cells. Therefore, we examined the transactivation function of p53 in these cells. Our data show the failure to transactivate p53, suggesting that increased p53 protein in T antigen expressing cells is functionless at least for transcriptional activation. PMID- 10376801 TI - Zinc-alpha2-glycoprotein expression as a marker of differentiation in human oral tumors. AB - Zinc-alpha2-glycoprotein (Znalpha2gp) is a soluble major histocompatibility complex homolog widespread in body fluids and in glandular epithelia; the authors recently demonstrated its presence in stratified epithelia. Znalpha2gp has been associated with tumor differentiation in breast cancers and other carcinomas. We compare here its gene expression in histopathologically graded oral squamous cell carcinomas and in their perilesional normals. Znalpha2gp levels are higher in the controls than in the tumors, and higher in well-differentiated tumors than in poorly differentiated ones. Markers of oral epithelial maturation (keratin K13 and involucrin) are less simply related to tumor histology. PMID- 10376803 TI - Calcium-dependent self-association of annexin II: a possible implication in exocytosis. AB - Alveolar type II cells secrete lung surfactant through exocytosis of lamellar bodies. We previously showed that the annexin II tetramer (Anx IIt) mediates the fusion of lamellar bodies with liposomes. The present study examined the possible involvement of membrane proteins in this process. Pre-treatment of lamellar bodies with trypsin and alpha-chymotrypsin reduced Anx IIt-mediated membrane fusion. With the use of an Anx IIt-conjugated Sepharose column, three Anx IIt binding proteins with molecular weights of 67,000, 36,000 and 34,000 were isolated froM the Triton X-100 extract of bovine lung tissue membranes. These proteins were identified as annexins VI, II and IV by Western blot. The interaction of Anx IIt with annexins II and IV was confirmed by ligand blot assay. An EGTA-resistant membrane-bound annexin II was present in lung type II cells. Anx IIt preferentially hound to membranous annexin II compared with cytosolic annexin II of type II cells. With the use of immunofluorescence, annexin II was found to translocate from cytoplasm to plasma membranes in type II cells upon stimulation with phorbol 12-myristate 13-acetate. These results suggest that cytosolic annexin II may bind to membranous annexin II and form a protein-protein bridge to bring two membranes together. PMID- 10376802 TI - Cyclic ADP-ribose: a novel Ca2+-mobilising second messenger. AB - Cyclic ADP-ribose (cADPR) was discovered as a potent Ca2+-mobilising natural compound in sea urchin eggs. Recently, cADPR was reported to stimulate Ca2+ signalling in several higher eukaryotic cell systems (e.g., smooth and cardiac muscle cells, neuronal cells, adrenal chromaffin cells, macrophages, pancreatic acinar cells and T-lymphocytes). The following aspects of the role of cADPR as a Ca2+-mobilising second messenger are reviewed: coupling of metabolism of cADPR to stimulation of receptors in the plasma membrane, properties and pharmacology of Ca2+ release by cADPR and the involvement of cADPR in Ca2+ entry. PMID- 10376804 TI - p42/p44 mitogen-activated protein kinase activation is involved in prostaglandin F2alpha-induced interleukin-6 synthesis in osteoblasts. AB - Prostaglandin F2alpha (PGF2alpha) significantly induced p42/p44 mitogen-activated protein (MAP) kinase activity in osteoblast-like MC3T3-E1 cells. PD98059, a selective inhibitor of MAP kinase kinase, inhibited PGF2alpha-induced interleukin 6 (IL-6) synthesis as well as PGF2alpha-induced p42/p44 MAP kinase activation. PD98059 suppressed the IL-6 synthesis induced by 12-O-tetradecanoylphorbol-13 acetate (TPA), a protein kinase C (PKC) activator, or NaF, an activator of heterotrimeric GTP-binding protein, as well as the p42/p44 MAP kinase activation by TPA or NaF. Calphostin C, a highly potent and specific inhibitor of PKC, inhibited the PGF2alpha-induced p42/p44 MAP kinase activity. These results strongly suggest that PKC-dependent p42/p44 MAP kinase activatioin is involved in PGF2alpha-induced IL-6 synthesis in osteoblasts. PMID- 10376805 TI - Role of STAT5 in interferon-alpha signal transduction in Ba/F3 cells. AB - In interferon-alpha (IFN-alpha) signalling, the essential role of the transcription factors STAT1 and STAT2 is well established. In contrast, the involvement of other STAT proteins, including STAT5, is much less well understood. Here we show that, in IFN-alpha-responsive Ba/F3 cells, this cytokine stimulates the DNA-binding of STAT5A and B but that IL-3 is a much more potent activator of both STAT5 isoforms. A stably expressed dominant-negative mutant of JAK2 suppressed the IL-3- but not the IFN-alpha-dependent DNA binding of STAT5, suggesting independent mechanisms of its activation. Northern blots revealed that IL-3 strongly induced the expression of two STAT5-regulated genes, pim-1 and oncostatin-M, whereas IFN-alpha had a weak stimulatory effect on pim-1 expression only. In summary our results suggest that, despite the capability of IFN-alpha to stimulate DNA binding of STAT5, this transcription factor does not play a pivotal role in IFN-alpha signalling in Ba/F3 cells. PMID- 10376806 TI - Coupling of beta-adrenergic receptors to cardiac L-type Ca2+ channels: preferential coupling of the beta1 versus beta2 receptor subtype and evidence for PKA-independent activation of the channel. AB - Beta1- and beta2-adrenergic receptors (beta-ARs) co-exist in mammalian heart, and it is generally accepted that both activate adenylyl cyclase (AC), resulting in increased levels of cAMP and subsequent activation of L-type Ca2+ channels (CaCh). To investigate the contribution of each beta-AR subtype in AC and CaCh coupling, we stably expressed cardiac CaCh alpha1 and beta2 subunits along with either beta1-AR or beta2-AR in CHW fibroblasts. Co-expression of either beta-AR with CaCh subunits conferred responsiveness of AC and CaCh to isoproterenol (ISO), which was not observed in non-transfected cells. ISO-promoted cAMP formation occurred at a lower EC50 through the beta2-AR than through the beta1-AR (0.13 +/- 0.01 vs. 0.6 +/- 0.14 nM). In contrast, activation of CaCh was more efficacious via the beta1-AR than the beta2-AR (EC50 for CaCh activation = 238 +/ 33 vs. 1057 +/- 113 nM). Pre-treatment with pertussis toxin (PTX) had no effect upon the responsiveness of either cAMP formation or CaCh activation through either receptor. We conclude (1) that beta1-ARs exhibit preferential coupling to CaCh activation, versus that observed for the beta2-AR; (2) that this preferential coupling cannot be explained solely by cAMP-dependent processes; and (3) that the relative attenuation of beta2-AR-promoted CaCh activation is not due to receptor coupling to PTX-sensitive G proteins. Thus, it is likely that other subtype-specific, cAMP-independent coupling of the beta-AR to CaCh is present. PMID- 10376807 TI - Role of voltage-dependent Na+ channels for rhythmic Ca2+ signalling in glucose stimulated mouse pancreatic beta-cells. AB - The putative role of voltage-dependent Na+ channels for glucose induction of rhythmic Ca2+ signalling was studied in mouse pancreatic beta-cells with the use of the Ca2+ indicator fura-2. A rise in glucose from 3 to 11 mM resulted in slow oscillations of the cytoplasmic Ca2+ concentration ([Ca2+]i). These oscillations, as well as superimposed transients seen during forskolin-induced elevation of cAMP, remained unaffected in the presence of the Na+ channel blocker tetrodotoxin. During exposure to 1-10 microM veratridine, which facilitates the opening of voltage-dependent Na+ channels, the slow oscillations were replaced by repetitive and pronounced [Ca2+]i transients arising from the basal level. The effects of veratridine were reversed by tetrodotoxin. The veratridine-induced [Ca2+]i transients were critically dependent on the influx of Ca2+ and persisted after thapsigargin inhibition of the endoplasmic reticulum Ca2+-ATPase. Both tolbutamide and ketoisocaproate mimicked the action of glucose in promoting [Ca2+]i transients in the presence of veratridine. It is suggested that activation of voltage-dependent Na+ channels is a useful approach for amplifying Ca2+ signals for insulin release. PMID- 10376809 TI - Intracellular free calcium regulates the onset of the respiratory burst of human neutrophils activated by phorbol myristate acetate. AB - Thapsigargin was used to study the regulation of different static calcium level ([Ca2+]i) on the respiratory hurst of human neutrophils stimulated with phorbol myristate acetate (PMA). The result showed that the onset time of the respiratory hurst was obviously reduced by elevation of static [Ca2+]i but is still much longer than that stimulated with N-formylmethionylleucylphenylalanine (fMLP). To find the reason, the onset times of the respiratory burst stimulated with fMLP, 1,2-dioctanoyl-sn-glycerol (DiC8), and PMA were determined at different static [Ca2+]i. It turns out that although DiC8 was unable to induce the respiratory burst at low [Ca2+], the onset time of DiC8-stimulated response at high [Ca2+]i was almost the same as that stimulated with fMLP. The study revealed that the fast onset of the fMLP-stimulated respiratory burst in comparison with PMA stimulated response is not only due to the transient rise of [Ca2+]i, but is also due to the higher efficiency of diacylglycerol (DAG) in activating protein kinase c (PKC). The determining step in governing the onset of a respiratory burst is the activation of PKC. PMID- 10376808 TI - Assessment of the extracellular and intracellular actions of sphingosine 1 phosphate by using the p42/p44 mitogen-activated protein kinase cascade as a model. AB - We have investigated the extracellular and intracellular actions of sphingosine 1 phosphate (S1P) by using cultured airway smooth muscle cells. We have demonstrated that exogenous S1P elicited an activation of mitogen-activated protein kinase (p42/p44 MAPK) that was abolished by pertussis toxin (0.1 microg/mL, 24 h), which was used to inactivate Gi. The effect of exogenous S1P might therefore be attributed to an action at a putative Gi-coupled receptor. The regulation of the p42/p44 MAPK cascade by S1P was also shown to include a protein kinase C (PKC)-dependent intermediate step. Platelet-derived growth factor (PDGF) stimulates intracellular S1P formation and was therefore used to evaluate the intracellular action of S1P. This has previously been investigated by others using the sphingosine kinase inhibitors D,L-threo-dihydrosphingosine and N,N dimethylsphingosine. We have demonstrated here that both inhibitors block the PDGF-dependent activation of p42/p44 MAPK. However, both are also PKC inhibitors, which might account for their effect because PDGF utilises PKC as an intermediate in the regulation of the p42/p44 MAPK cascade. Significantly, sphingosine, which is the substrate of sphingosine kinase and a PKC inhibitor, blocked the activation of p42/p44 MAPK by PDGF with an almost identical concentration dependence compared with D,L-threo-dihydrosphingosine and N,N dimethylsphingosine. Therefore, the use of so-called sphingosine kinase inhibitors might lead to misleading interpretations because of their additional effect on PKC. Other approaches, such as oligodeoxynucleotide anti-sense against sphingosine kinase, are required to address the intracellular role of S1P. PMID- 10376810 TI - 5-Hydroxytryptamine-induced phosphoinositide hydrolysis and Ca2+ mobilisation in canine cultured aorta smooth muscle cells. AB - The effect of 5-hydroxytryptamine (5-HT) on phospholipase C (PLC)-mediated phosphoinositide (PI) hydrolysis and intracellular Ca2+ ([Ca2+]i) changes was investigated in canine cultured aorta smooth muscle cells (ASMCs). 5-HT stimulated inositol phosphate (IP) accumulation was time and concentration dependent with a half-maximal response (pEC50) and a maximal response at 6.4 and 10 microM, n = 6, respectively. Stimulation of ASMCs by 5-HT produced an initial transient peak followed by a sustained, concentration-dependent elevation in [Ca+]i. The half-maximal response (pEC50) values of 5-HT for the peak and sustained plateau were 7.1 and 6.9, respectively. Ketanserin and mianserin (1 and 3 nM), 5-HT2A antagonists, were equipotent and had high affinity in antagonising the 5-HT-induced IP accumulation and [Ca2+]i change with pK(B) values of 8.6-9.1 and 8.6-9.4, respectively. In contrast, the concentration-effect curves of 5-HT induced IP and [Ca2+]i responses were not shifted until the concentrations of NAN 190 and metoctopramide (5-HT1A and 5-HT3 receptor antagonists, respectively) were increased to as high as 1 microM with pK(B) values of 5.7-6.3 and 6.1-6.6, respectively, indicating that the 5-HT receptor-mediated responses had low affinity for these antagonists. Pre-treatment of ASMCs with pertussis toxin (100 ng/mL, 24 h) caused a significant inhibition of 5-HT-induced IP accumulation and [Ca2+]i change in ASMCs. Depletion of external Ca2+ or removal of Ca2+ by addition of EGTA led to a significant attenuation of IP accumulation and [Ca2+]i change induced by 5-HT. Influx of external Ca2+ was required for the 5-HT-induced responses, because Ca2+-channel blockers--verapamil, nifedipine and Ni2+--partly inhibited the 5-HT-induced IP accumulation and Ca2+ mobilisation. The sustained elevation of [Ca2+]i response to 5-HT was dependent on the presence of external Ca2+. Removal of external Ca2+ by addition of 5 mM EGTA during the sustained phase caused a rapid decline in [Ca2+]i to lower than the resting level. The sustained elevation of [Ca2+]i could then be evoked by addition of 1.8 mM Ca2+ in the continued presence of 5-HT. These results demonstrate that 5-HT directly stimulates PLC-mediated PI hydrolysis and Ca2+ mobilisation, at least in part, through a pertussis toxin-sensitive G protein in canine ASMCs. 5-HT2A receptors may be predominantly mediating IP accumulation, and subsequently IP-induced Ca2+ mobilisation may function as the transducing mechanism for 5-HT-stimulated contraction of aorta smooth muscle. PMID- 10376812 TI - A hybrid response regulator possessing a PEP-dependent phosphorylation domain. PMID- 10376811 TI - Dual activity of pyrrolidine dithiocarbamate on kappaB-dependent gene expression in U937 cells: II. Regulation by tumour necrosis factor-alpha. AB - In the human promonocytic U937 cell line, pyrrolidine dithiocarbamate (PDTC) was a potent inhibitor of the nuclear factor-kappaB (NF-kappaB) signalling pathway induced by the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA). However, PDTC did not inhibit tumour necrosis factor-alpha (TNF-alpha)-induced NF-kappaB DNA binding activity but potentiated the effect of TNF-alpha on kappaB-dependent gene expression. The stimulatory effect of PDTC with TNF-alpha was not observed with an HIV-1 LTR reporter construct containing two mutated kappaB binding sites or with a construct with a mutation of the activating protein (AP)-2 binding site located between the two kappaB elements. Two distinct signalling pathways, one mediated by TPA and the other by TNF-alpha, were shown to interact, functionally defining a threshold important in the inhibitory or stimulatory effect of PDTC on kappaB-dependent gene expression. Evidence that PDTC induced AP-1 DNA binding and AP-1 reporter gene activity, raised the hypothesis that the effect of PDTC was mediated by an interaction between the AP-1 pathway and p65(RelA). Co transfection with expression vectors for p65(RelA) and the AP-1 subunits c-Fos and c-Jun resulted in a decrease in the stimulatory effect of PDTC on HIV-1 LTR activity. Co-transfection of p65(RelA) with Tam67, a dominant negative mutant of c-Jun defective in transactivation, stimulated the effect of PDTC on HIV-1 LTR activity. Evidence that the stimulatory effect of Tam67 with PDTC was reduced with c-Jun is consistent with the hypothesis. PMID- 10376813 TI - Protein targeting to the endoplasmic reticulum in yeast. 1997 Fleming Lecture. PMID- 10376814 TI - Proteolytic cleavage of the A subunit is essential for maximal cytotoxicity of Escherichia coli O157:H7 Shiga-like toxin-1. AB - Members of the bacterial Shiga toxin family consist of a single A subunit that is non-covalently associated with a pentamer of B subunits. These toxins bind to receptors on susceptible mammalian cells and enter the cells by endocytic uptake. During cell entry, the 32 kDa A subunit is cleaved by the membrane-anchored protease furin to generate a catalytically active, 27.5 kDa A1 fragment and a 4.5 kDa A2 fragment. Previous studies have shown that mutating the furin site to prevent cleavage did not significantly affect toxin potency, suggesting that cleavage is not required for toxin activity. Here it is confirmed that preventing cleavage at the usual processing site does not prevent proteolytic processing of the Escherichia coli Shiga-like toxin-1 A subunit. However, simultaneous mutation of both the primary furin-recognition site and a nearby putative furin cleavage site did prevent intracellular processing of the A subunit. Comparison of the cytotoxicities of purified recombinant toxins to cultured mammalian cells demonstrated that even on prolonged incubation with toxin, the unprocessed mutant was 60-fold less toxic than the wild-type protein or other mutants still capable of being proteolytically processed during cell entry. PMID- 10376815 TI - The large plasmids of Shiga-toxin-producing Escherichia coli (STEC) are highly variable genetic elements. AB - Shiga-toxin-producing Escherichia coli (STEC) of different serotypes are known to harbour large plasmids. The aim of this study was to investigate, using the example of the plasmid-encoded serine protease EspP, whether these plasmids are a uniform genetic element present in STEC. Examination of 201 diarrhoeagenic E. coli strains using a newly developed espP-specific PCR showed that espP is specific for STEC and present in 57% of STEC belonging to 16 different serotypes. The espP genes of the 16 STEC serotypes varied to a certain extent, as shown by nucleotide sequence and restriction enzyme analyses, but the DNA regions adjacent to the espP gene were completely different. When two further STEC-plasmid markers, the catalase-peroxidase gene katP and the enterohaemorrhagic E. coli haemolysin gene EHEC-hlyA were included, many combinations of the three markers were found, depending in part on the serotype. In addition, strains possessing none of the three markers still harboured large plasmids. In the most prevalent STEC serogroup, O157, it was observed that the plasmid of sorbitol-fermenting STEC O157:H- lacks the espP and katP genes although both genes are present in the plasmid of the non-sorbitol-fermenting STEC O157:H7. The EHEC-hlyA gene, however, is present in both. In conclusion, this study shows that the large plasmids of STEC are not uniform genetic elements but heterogeneous in both their gene composition and arrangement. PMID- 10376816 TI - Varying division planes of secondary constrictions in spheroidal Escherichia coli cells. AB - Planes of successive divisions in Escherichia coli have been proposed to be either parallel or perpendicular to each other, restricted to one or two dimensions. To test the hypothesis that divisions can occur in planes alternating in three dimensions, a method was developed to generate cells with secondary constrictions during growth in suspension. The method involves a combination of thymine limitation (to manipulate chromosome replication rate) and mecillinam treatment (to inhibit penicillin-binding protein 2). The former modifies timing of terminations, the latter results in spheroidal cells. Such cells displayed secondary constrictions after adding deoxyguanosine (accelerating replication rate), thus temporarily enhancing division signals. The successive constrictions were seen to develop in planes that were tilted relative to each other, and in positions related to those of the nucleoids, visualized by staining with DAPI (4',6-diamidino-2-phenylindole dihydrochloride hydrate). Visualizing cell envelopes with FM 4-64 by confocal scanning laser microscopy supported the conclusion that planes of successive divisions can alternate in three dimensions. PMID- 10376817 TI - Fimbriae- and flagella-mediated association with and invasion of cultured epithelial cells by Salmonella enteritidis. AB - Salmonella enteritidis expresses flagella and several finely regulated fimbriae, including SEF14, SEF17 and SEF21 (type 1). A panel of mutants was prepared in three strains of S. enteritidis to elucidate the role of these surface appendages in the association with and invasion of cultured epithelial cells. In all assays, the naturally occurring regulatory-defective strain 27655R associated with tissue culture cells significantly more than wild-type progenitor strains LA5 and S1400/94. Compared with wild-type strains, SEF14 mutants had no effect on association and invasion, whereas SEF17, SEF21 and aflagellate mutants showed significant reductions in both processes. Histological examination suggested a role for SEF17 in localized, aggregative adherence, which could be specifically blocked by anti-SEF17 sera and purified SEF17 fimbriae. SEF21-mediated association was neutralized by mannose and a specific monoclonal antibody, although to observe enhanced association it was necessary for the bacteria to be in fimbriate phase prior to infection. Additionally, aflagellate mutants associated and invaded less than motile bacteria. This study demonstrated the potential for multifactorial association and invasion of epithelial cells which involved SEF17 and SEF21 fimbriae, and flagella-mediated motility. PMID- 10376818 TI - Structural analysis of Bacillus megaterium KM spore peptidoglycan and its dynamics during germination. AB - The composition and structure of peptidoglycan from dormant spores of Bacillus megaterium KM and its dynamics during germination were investigated. Amino acid analysis and mass spectrometry identified 21 muropeptides resolved by reverse phase HPLC following digestion of peptidoglycan with Cellosyl. The basic structure of peptidoglycan in B. megaterium spores is similar to that of Bacillus subtilis: 44.2% of muramic acid residues are substituted with delta-lactam, 28.8% with single L-alanine, 25.1% with tetrapeptide and only 1.8% with tripeptide. The cross-linking index of the spore peptidoglycan, determined from muropeptides resolved by reverse phase HPLC, was 2.2 % per muramic acid. Spore peptidoglycan contains 2.9% of muropeptides with unsubstituted N-acetylmuramic acid. These muropeptides are likely to be intermediate products of delta-lactam formation. Analysis of muropeptide dynamics during germination revealed the activity of at least two hydrolytic enzymes, an N-acetylglucosaminidase and a lytic transglycosylase. A 4 M LiCl extract from 30 min germinated spores of B. megaterium KM caused 'germination-like' changes to permeabilized spores of B. megaterium and B. subtilis but not those of a B. subtilis cwlD mutant. Muropeptide analysis of the treated spores revealed the presence of products generated by the activity of a glucosaminidase. PMID- 10376819 TI - Real-time monitoring of Bacillus subtilis endospore components by attenuated total reflection Fourier-transform infrared spectroscopy during germination. AB - Chemical changes of particular Bacillus subtilis spore components were monitored by attenuated total reflection Fourier-transform infrared spectroscopy (ATR/FTIR) during spore germination on a ZnSe internal reflection element. Within minutes of the initiation of spore germination, significant changes in the amount of calcium dipicolinate (DPA-Ca) and proteins were noted in the wild-type strain. The changes in a germination mutant (strain 1G9, gerD) were similar to those in the wild-type strain, but the rates of change were slower. The changes in another germination mutant (strain 1G7, gerA) were very different from those in the first two strains: germination was slow and incomplete, and proteins and DPA-Ca remained unaltered throughout the course of the germination study. This technique thus offers a sensitive and non-destructive method for real-time monitoring of various cellular components during spore germination. PMID- 10376820 TI - Assay for UDPglucose 6-dehydrogenase in phosphate-starved cells: gene tuaD of Bacillus subtilis 168 encodes the UDPglucose 6-dehydrogenase involved in teichuronic acid synthesis. AB - A novel assay permitting the detection of UDPglucose 6-dehydrogenase activity in cell-free extracts obtained from phosphate-starved cultures of Bacillus subtilis is described. The critical step, the separation of phosphate-starvation-induced exo-enzymes, phosphatases and phosphodiesterases from the cytoplasmic fraction containing the UDPglucose dehydrogenase, was achieved by protoplasting and removal of the periplasmic fraction by protoplast washing. Using this method, the following were unambiguously demonstrated: (i) the presence in the cytoplasm of an enzymic activity oxidizing UDPglucose to UDPglucuronic acid, and (ii) that detection of the activity in whole-cell-free extracts is prevented by the presence of 'periplasmic' enzymes catalysing the degradation of the sugar nucleotides. With this method, several B. subtilis 168 mutants unable to synthesize teichuronic acid were examined. Strains inactivated in gene tuaD, whose product shares homology with UDPglucose 6-dehydrogenase and GDPmannose 6 dehydrogenase from other organisms, were shown to lack UDPglucose 6-dehydrogenase activity. Anion exchange chromatography revealed that mutants deficient in tuaD lacked a cytoplasmic UDPglucuronate pool. PMID- 10376821 TI - Nucleotide sequence of the Bacillus subtilis temperate bacteriophage SPbetac2. AB - The Bacillus subtilis 168 chromosomal region extending from 184 degrees to 195 degrees, corresponding to prophage SPbeta, has been completely sequenced using DNA of the thermoinducible SPbetac2 mutant. This 134416 bp segment comprises 187 putative ORFs which, according to their orientation, were grouped into three clusters. Compared to its host, SPbetac2 is characterized by a lower G+C content, shorter mean ORF length, as well as a different usage of start codons. Nearly 75% of predicted ORFs do not share significant homologies to sequences in available databases. The only highly similar proteins to SPbetac2-encoded ones are host paralogues. SPbetac2 promoter regions contain SOS box consensus sequences and a repeated motif, designated SPbeta repeated element (SPBRE), that is absent from the host genome. Gene sspC, encoding the small acid-soluble protein C, that has been previously sequenced and mapped to the vicinity of the SPbeta region, was found to be part of the prophage. PMID- 10376822 TI - Two genes from Bacillus subtilis under the sole control of the general stress transcription factor sigmaB. AB - The general stress response of Bacillus subtilis is triggered by a variety of environmental and metabolic stresses which activate the sigmaB transcription factor. Among the more than 100 genes controlled by sigmaB (the csb genes), the functions identified thus far include resistance to oxidative stress, resistance to protein denaturation and resistance to osmotic stress. To understand the breadth of functions in which csb genes participate, the transcriptional organization and predicted products of two such genes previously identified in a screen for sigmaB-dependent lacZ fusions were analysed. The csb-22::Tn917lacZ and csb-34::Tn917lacZ fusions are unusual among csb genes in that their expression appears to be completely dependent upon sigmaB. By plasmid-integration experiments, fusion analyses and site-directed mutagenesis, stress-inducible, sigmaB-dependent promoters for both these fusions were identified. The csb-34 fusion marked an ORF (yxcC or csbC) which by sequence analysis lay in a monocistronic transcriptional unit. This ORF encoded a predicted 461-residue product which had high identity with Class I sugar transporters of the major facilitator superfamily. It was speculated that the csbC product could serve either a nutritional or an osmotic protection function. In contrast, the csb-22 fusion identified an ORF (ywmG or csbD) which appeared to be the second gene of a two-gene operon. This ORF encoded a predicted 62-residue product which resembled a small Escherichia coli protein of unknown function. The sigmaB. dependent promoter lay immediately upstream from csbD and appeared to be an internal promoter for the operon. PMID- 10376823 TI - Inactivation of the KIPMR1 gene of Kluyveromyces lactis results in defective cell wall morphogenesis. AB - The P-type Ca2+ -ATPases are the transporters responsible for calcium homeostasis in the cell compartments of eukaryotes. The KIPMR1 gene of Kluyveromyces lactis encodes a P-type Ca2+ -ATPase, which is functionally and structurally homologous to Pmr1p of Saccharomyces cerevisiae, the calcium pump localized in the Golgi membranes. In this work, a novel involvement of KIPmr1p in cell-wall morphogenesis of K. lactis is reported. KIpmr1delta cells exhibited the loss of outer-chain extension in the glycosylation of secreted proteins. The absence of KIPmr1p resulted in the accumulation of round, large cells with an abnormally thick cell wall, as revealed by transmission electron microscopy. The deletant strain also showed a delocalized deposition of chitin in the lateral cell wall accompanied by an unbalanced ratio of insoluble to soluble glucans. These morphological defects were accompanied by the presence of irregularly shaped nuclei and by a DNA content greater than 2n. Addition of 10 mM Ca2+ to the medium of the KIpmr1delta strain reversed the chitin-deposition impairment, recovered the alteration to the glucan ratio and restored a normal thickness of the cell wall. The mutant cells resumed wild-type size, shape and nuclear morphology but the DNA content indicated the persistence of defects in the co-ordination between DNA replication and cell division. The glycosylation defects were completely unaffected by the calcium supplement. These results indicate that calcium homeostasis controlled by KIPmr1p plays an important role in the cell-wall morphogenesis of K. lactis. PMID- 10376824 TI - A unique eukaryotic beta-xylosidase gene from the phytopathogenic fungus Cochliobolus carbonum. AB - The plant-pathogenic fungus Cochliobolus carbonum secretes one major beta xylosidase (Xyp1) when grown on xylan or maize cell walls. cDNA and genomic DNA encoding Xyp1 were isolated using PCR primers based on peptide sequences from the purified protein. XYP1 contains three introns, has 5' and 3' untranslated regions of 74 and 145 bp, respectively, and is predicted to encode a protein of 328 amino acids (Mr 36700) with four N-glycosylation sites. Although it is secreted, Xyp1 has no predicted signal peptide. Furthermore, Xyp1 appears not to be processed at the N-terminus because one of the peptides isolated from the mature protein is located only six amino acids downstream of the translational start methionine. The primary sequence of Xyp1 is unrelated to any known eukaryotic beta-xylosidase but has 35% overall identity to two bacterial bifunctional beta-xylosidase/alpha arabinosidases. Mutation of XYP1 by targeted gene replacement resulted in the loss of the major beta-xylosidase activity corresponding to the product of XYP1, but a significant amount of secreted beta-xylosidase activity (25% of wild-type) remained in the culture filtrates. The xyp1 mutant was still fully pathogenic on maize. PMID- 10376825 TI - The small GTP-binding protein Rho is expressed differentially during spore germination of Phycomyces blakesleeanus. AB - Evidence has been obtained for the presence of the small 22 kDa GTP-binding Rho protein in dormant spores of Phycomyces blakesleeanus. Immunoblotting with a polyclonal antibody against RhoA revealed a soluble and membrane-associated 22 kDa protein. When [32P]ADP-ribosylated by Clostridium botulinum C3 exotoxin the protein had a pI of 5.7, a value similar to that reported for other RhoA proteins. The 22 kDa protein was expressed at all stages of growth investigated, but radiolabelling of the [32P]ADP-ribosylated protein increased when tube formation occurred and decreased as the hyphae branched. Localization of RhoA during spore germination studied by immunofluorescence microscopy revealed the presence of RhoA in the cell membrane of the spore. When the spore started to swell, RhoA was observed as patches in the cell membrane which become concentrated in the neck region of the site of the protuberation tube, but this protein was never observed at the point of growth of the hyphal tip. The above results suggest that RhoA associated with one or more membrane proteins could participate in the molecular mechanism involved in maintaining cell integrity during the extrusion of the germ-tube of P. blakesleeanus. PMID- 10376826 TI - Different temporal and spatial expression of two hydrophobin-encoding genes of the edible mushroom Agaricus bisporus. AB - In a search for genes that are only expressed in fruit bodies of the basidiomycete Agaricus bisporus, two cDNAs, hypA and hypB that encode hydrophobins have been isolated previously. In this study, the structure of hypB is resolved and it is shown that the two genes are differentially expressed, indicating that the encoded hydrophobins serve different functions in A. bisporus mushrooms. hypB encodes a polypeptide (HYPB) of 119 aa that shows little sequence identity with HYPA apart from the characteristic arrangement of eight cysteines found exclusively in hydrophobins. The temporal and spatial expression of the two hydrophobin-encoding genes during fruit body development was compared using Northern analysis and in situ hybridization. Accumulation of hypA mRNA was found in tissue fractions consisting of undifferentiated white hyphae. In situ hybridization showed that the highest hypA mRNA levels are not found in the outermost cell layers of the pileipellis but in the cell layers adjacent to that. The highest level of expression of hypB occurs early in development when the primordium differentiates into densely packed, randomly oriented cap hyphae and loosely packed, vertically oriented stipe hyphae. In mature mushrooms, a strong accumulation of hypB transcripts was found only in the transitional zone between cap and stipe tissue, demonstrating that transcription regulation of hypB is clearly distinct from hypA. PMID- 10376827 TI - Changes in Aspergillus nidulans gene expression induced by bafilomycin, a Streptomyces-produced antibiotic. AB - In natural environments bacteria and filamentous fungi often compete for the same resources. Consequently, production of antibiotic secondary metabolites and defence mechanisms against these compounds have evolved in these organisms. An experimental model has been developed to study the response in fungi exposed to one such antibiotic. The filamentous fungus Aspergillus nidulans was treated with bafilomycin B1, a Streptomyces-produced antibiotic which reduces radial growth rate and induces morphological changes in fungi. mRNA differential display was used to study changes in fungal gene expression. For five genes, changes in abundance of the corresponding mRNAs, directly or indirectly caused by bafilomycin, were observed. Of these, three were up-regulated and two repressed. With four of these the change in mRNA abundance measured ranged from 10- to 60 fold. However, for one gene the mRNA was only detected after bafilomycin treatment. One of the down-regulated mRNAs encodes ASPND1, a glycoprotein that belongs to a known family of antigens identified in aspergilloma patients. One up regulated mRNA shows sequence similarities, at the amino acid level, with a cell wall protein of Saccharomyces cerevisiae. The remaining three genes were also cloned and sequenced; their sequences do not correspond to known genes in A. nidulans, and no similarities with published nucleotide or protein sequences in other organisms were found. These results indicate the feasibility of using mRNA differential display to study interactions between bacteria and filamentous fungi. PMID- 10376828 TI - Purification of geranylgeranyltransferase I from Candida albicans and cloning of the CaRAM2 and CaCDC43 genes encoding its subunits. AB - All previously characterized protein geranylgeranyltransferases I (GGTase I) are heterodimeric zinc metalloenzymes which catalyse geranylgeranylation of a cysteine residue in proteins containing a C-terminal CaaL motif (C, Cys; a, aliphatic amino acid; L, Leu). The alpha and beta subunits of GGTase I of Saccharomyces cerevisiae are encoded by RAM2 and CDC43, respectively, and are essential for yeast viability. The authors are therefore investigating the role of geranylgeranylation in the related pathogenic yeast, Candida albicans, which is the most prevalent human fungal pathogen. GGTase I was purified to near homogeneity and also found to be a heterodimeric magnesium-dependent, zinc metalloenzyme displaying selectivity for CaaL-containing protein substrates. GGTase I peptide sequences were obtained from the purified protein and used to clone the genes encoding both subunits. CaRAM2 and CaCDC43 encode proteins that are 42 and 34% identical to their corresponding S. cerevisiae homologues, respectively, and 30% identical to their human homologues. Despite the limited overall homology, key zinc- and substrate-binding residues of the beta subunit (Cdc43p) are conserved. A unique feature of CaCdc43p is a tract of polyasparagine whose length varies from 6 to 17 residues among C. albicans strains and between alleles. Coexpression of both CaCDC43 and CaRAM2 under their native promoters complemented the ts defect of a S. cerevisiae cdc43 mutant but expression of the beta-subunit alone did not correct the growth defect, suggesting that hybrid GGTase I heterodimers are nonfunctional. PMID- 10376829 TI - Towards understanding the evolution of the human commensal yeast Candida albicans. AB - Allelic frequencies and relationships for one dimorphic locus and three unlinked polymorphic loci have been determined for 114 unrelated isolates of Candida albicans, including 14 laboratory reference strains and 50 strains from each of two geographic regions. Although there was no indication of geographical partitioning, there were significant correlations for specific allelic pairs among loci and little evidence that any alleles were in Hardy-Weinberg equilibrium. This gives additional support for the concept that the primary mode of genetic inheritance in this species is clonal, with other intracellular genetic events playing a lesser role in the creation of genomic diversity. Through inference of this and other known attributes of closely related Candida species, such as sequence analysis of IS1 and the ITS2 (internal transcribed spacer 2) region of the rDNA cistron, the deduced phylogeny suggests an evolutionarily recent origin for many frequently isolated strains. This finding will be of interest in the context of understanding pathogenicity and drug resistance in this human commensal yeast. PMID- 10376830 TI - Involvement of outer-membrane proteins in the aggregation of Azospirillum brasilense. AB - A bioassay was developed to investigate biological factors involved in the aggregation of Azospirillum brasilense strain Cd. Cells were grown for 24 h under aggregation-inducing and non-aggregation-inducing conditions (high and low C:N, respectively) and sonicated for 20 s. The cells were washed by centrifugation and resuspended in potassium phosphate buffer containing the two types of sonication extract. A greater extent of aggregation and higher flocculation were observed after 2-3 h incubation in the presence of sonicates from cells grown at high C:N (H-cells) compared to cells grown at low C:N. Flocculation did not occur after incubation of these cells in phosphate buffer. Boiled or proteinase K-treated sonicates originating from H-cells had lower aggregation-inducing capacity. After fractionation of the crude sonicate, both the outer-membrane protein (OMP) and the total membrane (mostly OMP) fractions possessed relatively high aggregation specific activities. The aggregation-inducing capacity of the OMP fraction strongly correlated with its protein concentration in the bioassay. Treatment of this fraction with proteinase K also decreased its aggregation-inducing activity. These findings suggest that OMPs are involved in the aggregation process of cells of A. brasilense. PMID- 10376831 TI - Anaerobic oxidations of cysteate: degradation via L-cysteate:2-oxoglutarate aminotransferase in Paracoccus pantotrophus. AB - Anoxic, fresh-water enrichment cultures to oxidize different organosulfonates were set up with nitrate, ferric iron or sulfate as electron acceptors. Pure cultures were easily obtained with two naturally occurring sulfonates, cysteate (2-amino-3-sulfopropionate) and taurine (2-aminoethanesulfonate), under nitrate reducing conditions. These two sulfonates were also oxidized during reduction of iron(III), though isolation of pure cultures was not successful. One nitrate reducing cysteate-oxidizing bacterium, strain NKNCYSA, was studied in detail. It was identified as Paracoccus pantotrophus. Eighteen sulfonates were tested, and the organism degraded cysteate, taurine, isethionate (2-hydroxyethanesulfonate), sulfoacetate or 3-aminopropanesulfonate with concomitant reduction of nitrate, presumably to molecular nitrogen. The carbon skeleton of these substrates was converted to cell material and, presumably, CO2. The amino group was released as ammonia and the sulfono moiety was recovered as sulfate. Cell-free extracts of P. pantotrophus NKNCYSA contained constitutive L-cysteate:2-oxoglutarate aminotransferase (EC 2.6.1.-) and glutamate dehydrogenase (EC 1.4.1.4). Taurine:pyruvate aminotransferase, in contrast, was inducible. PMID- 10376833 TI - Butane metabolism by butane-grown 'Pseudomonas butanovora'. AB - The pathway of butane metabolism by butane-grown 'Pseudomonas butanovora' was determined to be butane --> 1-butanol --> butyraldehyde butyrate. Butane was initially oxidized at the terminal carbon to produce 1-butanol. Up to 90% of the butane consumed was accounted for as 1-butanol when cells were incubated in the presence of 5 mM 1-propanol (to block subsequent metabolism of 1-butanol). No production of the subterminal oxidation product, 2-butanol, was detected, even in the presence of 5 mM 2-pentanol (an effective inhibitor of 2-butanol consumption). Ethane, propane and pentane, but not methane, were also oxidized. Butane-grown cells consumed 1-butanol and other terminal alcohols. Secondary alcohols, including 2-butanol, were oxidized to the corresponding ketones. Butyraldehyde was further oxidized to butyrate as demonstrated by blocking butyrate metabolism with 1 mM sodium valerate. Butyrate also accumulated from butane when cells were incubated with 1 mM sodium valerate. The pathway intermediates (butane, 1-butanol, butyraldehyde and butyrate) and 2-butanol stimulated O2 consumption by butane-grown cells. 1-Butanol, butyraldehyde and butyrate supported growth of 'P. butanovora', as did 2-butanol and lactate. PMID- 10376832 TI - Possible involvement of cAMP in aerial mycelium formation and secondary metabolism in Streptomyces griseus. AB - In Streptomyces griseus, A-factor (2-isocapryloyl-3R-hydroxymethyl-gamma butyrolactone) triggers secondary metabolism and morphogenesis by binding a repressor protein (ArpA) and dissociating it from DNA. UV-mutagenesis of the A factor-deficient mutant HH1 generated strain HO2, defective in the synthesis of ArpA and therefore able to form aerial mycelium, spores and streptomycin. Shotgun cloning of chromosomal DNA from wild-type S. griseus in strain HO2 yielded a gene that suppressed aerial mycelium formation and streptomycin production. Nucleotide sequencing and subcloning revealed that the gene encoded a eukaryotic-type adenylate cyclase (CyaA). In mutant HO2 production of cAMP was growth-dependent until the middle of the exponential growth stage; the production profile was the same as in the wild-type strain. However, the amount of cAMP produced was five times larger when mutant HO2 harboured cyaA on the high-copy-number plasmid pIJ486. Consistent with this, supplying cAMP exogenously at a high concentration to mutant HO2 suppressed formation of both aerial mycelium and streptomycin. On the other hand, some lower concentrations of cAMP stimulated or accelerated aerial mycelium formation. No effects of exogenous cAMP on morphogenesis and secondary metabolism were apparent in the wild-type strain. In addition, disruption of the chromosomal cyaA gene in the wild-type strain had almost no effect. Introducing cyaA cloned in either a low- or a high-copy-number plasmid suppressed morphogenesis and secondary metabolism not only in mutant HO2 but also in other arpA mutants, implying that the effects of cAMP became apparent in the arpA-defective background. When mutant HO2 carried cyaA on a plasmid, synthesis of the stringent response factor ppGpp was greatly reduced; this may account for the observed suppression by cAMP of morphogenesis and secondary metabolism. cAMP also affected protein tyrosine phosphorylation, as determined with antiphosphotyrosine antibody. PMID- 10376834 TI - Yersiniabactin from Yersinia pestis: biochemical characterization of the siderophore and its role in iron transport and regulation. AB - A siderophore-dependent iron transport system of the pathogenic yersiniae plays a role in the pathogenesis of these organisms. The structure of the yersiniabactin (Ybt) siderophore produced by Yersinia enterocolitica has been elucidated. This paper reports the purification of Ybt from Yersinia pestis and demonstrates that it has the same structure as Ybt from Y. enterocolitica. Purified Ybt had a formation constant for Fe3+ of approximately 4x10(-36). Addition of purified Ybt from Y. pestis enhanced iron uptake by a siderophore-negative (irp2) strain of Y. pestis. Maximal expression of the Ybt outer-membrane receptor, Psn, in this strain was dependent upon exogenously supplied Ybt. Regulation of Psn expression by Ybt occurred at the transcriptional level. Y. pestis DNA was used to construct irp2 and psn mutations in Yersinia pseudotuberculosis. The irp2 mutant strain no longer synthesized Ybt and the psn mutant strain could not use exogenously supplied Ybt. As in Y. pestis, Ybt was required for maximal expression of Psn. Regulation by Ybt occurred at the transcriptional level. In contrast to Y. pestis, in which a psn mutation does not repress synthesis of Ybt siderophore or expression of the iron-regulated HMWP1 and HMWP2 proteins, the same mutation in Y. pseudotuberculosis partially repressed these products. PMID- 10376835 TI - Characterization of glycolipids from Meiothermus spp. AB - Thin-layer chromatographic analysis of the polar lipids of Meiothermus strains revealed two glycolipid bands with similar chromatographic mobility to the major glycolipid of Thermus strains. In this study the glycolipids from the type strains of Meiothermus ruber, Meiothermus chliarophilus, Meiothermus silvanus and Meiothermus cerbereus were characterized using GC, GC/MS, fast atom bombardment MS and chemical methods. All strains contained dihexosyl-(N acyl)hexosaminylglucosyl diacylglycerols, related in structure to the major glycolipid of Thermus strains but varying in their fatty acylation pattern. The detection of two glycolipid bands by TLC in Meiothermus spp. was attributable to the invariable presence of 2-hydroxyacyl groups N-linked to the hexosamine of the polar head group which cause the glycolipids to be more strongly retained on silica TLC plates than 3-hydroxy or non-hydroxylated N-acyl glycolipids of similar structure that are also present. M. silvanus contained, in addition to these glyceroglycolipids, several glycolipids which were linked to acylated branched octadecanediols rather than to glycerol. The presence of glycolipids containing 2-hydroxyacyl groups N-linked to hexosamine appears to be a stable phenotypic marker that distinguishes the genus Meiothermus from the genus Thermus. PMID- 10376836 TI - Transposition of IS117 of Streptomyces coelicolor A3(2) in Mycobacterium smegmatis. AB - Derivatives of IS117, the Streptomyces coelicolor A3(2) 2.6 kb minicircle, transpose efficiently in Mycobacterium smegmatis, targeting chromosomal sites resembling translation start signals. Two IS117 derivatives, pIJ4696 and pIJ4697, containing a Streptomyces hygromycin-resistance gene in opposite orientations were introduced into M. smegmatis by electroporation and found to integrate into one of three specific sites. Integrations at sites A and B were frequent while integration at site C was observed only once. Only one site was occupied in each transformant. Sites A and B had either single or tandem integrations. PFGE analysis located these sites on different genomic Asel fragments. The sequences of the chromosome-IS117 junctions confirmed that integration was via the same IS117 attachment site as in Streptomyces, that there was no target site duplication, and that the orientation of IS117 at each site was fixed. In contrast to the situation in Streptomyces lividans, no deletions were created by the transposition and no circular forms could be detected. Comparison of the three M. smegmatis chromosomal 15117 target sites (attB) with known primary and secondary S. lividans attB sites showed that only a 2 bp 'AG' sequence at the crossover point was conserved. Dividing the attB sites into two groups produced two longer consensus target sites, GtcAAGg and gCCGATAGg. Most of the IS117 target sites resemble translational start sites, and site C resembles strongly the amino-terminal sequence of a Mycobacterium tuberculosis aminopeptidase. The level of hygromycin resistance in the transformants was high and independent of the site of integration, the number of copies integrated, or the orientation of the hyg gene. pIJ4696 at all three sites was stable in M. smegmatis in the absence of selection for at least 60 cell divisions. pIJ4696, pIJ4697 and other IS117 derivatives are promising vectors for the stable, integrative cloning of genes in M. smegmatis. PMID- 10376837 TI - The NucE and NucD lysis proteins are not essential for secretion of the Serratia marcescens extracellular nuclease. AB - The nuclease of Serratia marcescens is an extracellular protein encoded by the nucA gene. Pre-nuclease carries a typical 21-amino-acid N-terminal signal sequence that interacts with the Sec machinery to allow the translocation of nuclease to the periplasm. In Escherichia coli the nuclease remains in the periplasm; however, S. marcescens has the capacity to secrete nuclease extracellularly. The nucC operon carrying the nucEDC genes of S. marcescens has been identified previously. NucC is a transcriptional activator necessary for expression of nuclease as well as the extracellular bacteriocin 28b. NucE resembles and can act as a bacteriophage holin, whereas NucD has homology to bacteriophage lysozyme-like proteins. When present on a multicopy plasmid, the nucC operon, and specifically the nucED genes, appeared to allow extracellular secretion of nuclease from E. coli. Here experiments are reported which demonstrate that, when the nucC operon was placed in the E. coli chromosome in single copy, nuclease secretion was lost and nuclease remained periplasmic. The converse experiment, deletion of the nucE and nucD genes from the chromosome of S. marcescens, likewise had no effect on nuclease secretion by S. marcescens. It is concluded therefore that NucD and NucE are not necessary for nuclease secretion. PMID- 10376838 TI - gdhB, a gene encoding a second quinoprotein glucose dehydrogenase in Pantoea citrea, is required for pink disease of pineapple. AB - The pink disease of pineapple, caused by the bacterium Pantoea citrea, is characterized by a dark coloration on fruit slices after canning. A glucose dehydrogenase (Gdh) encoded by the gdhA gene has been implicated in the colour formation activity of P. citrea. In this paper it has been shown that P. citrea contains a second, homologous gdh gene and its product, GdhB, represents the main source of Gdh activity in this organism. Unlike gdhA, gdhB is constitutively expressed during the exponential phase of growth and is induced in stationary phase. A previously isolated chemical mutant, CMC6, which is deficient in Gdh activity and pink disease formation, failed to express gdhB during the stationary phase of growth. The CMC6 mutant can be complemented by a 54 bp DNA fragment located upstream of gdhA. This fragment, which contains an operator-like 11 bp inverted repeat, strongly enhances the expression of gdhA, probably by titrating away a negative effector of its expression. These results illustrate the complex interplay operating between the two gdh genes and emphasize the role of glucose metabolism in the pathway leading to pink disease. PMID- 10376839 TI - Effects of gene disruptions in the nisin gene cluster of Lactococcus lactis on nisin production and producer immunity. AB - The lantibiotic nisin is produced by several strains of Lactococcus lactis subsp. lactis. The chromosomally located gene cluster nisABTCIPRKFEG is required for biosynthesis, development of immunity, and regulation of gene expression. Inframe deletions in the nisB and nisT genes, and disruption of nisC by plasmid integration, eliminated nisin production and resulted in a strongly reduced level of immunity of the strains. The transcription of two nisin operons was inactivated in these mutant strains, but could be restored by addition of small amounts of nisin to growing cultures. The immunity levels of the mutants were also raised by adding nisin to growing cultures, albeit not to wild-type level. A strain with an in-frame deletion in the nisI gene was still able to produce active nisin, but the production and immunity levels were markedly lower. By measuring immunity levels of the knock-out strains and determining mRNA levels, it is concluded that NisI has an important function for nisin immunity and must cooperate with nisFEG-encoded proteins to provide a high level of immunity. Maximal immunity could not be obtained in the mutant strains, probably because the wild-type transcription levels from nisA and nisF promoters are not reached when essential nis genes are disrupted. Using Southern hybridization with a consensus promoter probe, no other DNA sequences similar to the nisA and nisF promoters could be detected, indicating that these two elements are probably the only ones in the chromosome regulated by nisin and are thus the only ones involved in the regulation of producer immunity. PMID- 10376840 TI - Role of multiple gene copies in particulate methane monooxygenase activity in the methane-oxidizing bacterium Methylococcus capsulatus Bath. AB - Genes for the subunits of particulate methane monooxygenase, PmoABC, have been sequenced from the gamma-proteobacterial methanotroph Methylococcus capsulatus Bath. M. capsulatus Bath contains two complete copies of pmoCAB, as well as a third copy of pmoC. The two pmoCAB regions were almost identical at the nucleotide sequence level, differing in only 13 positions in 3183 bp. At the amino acid level, each translated gene product contained only one differing residue in each copy. However, the pmoC3 sequence was more divergent from the two other pmoC copies at both the far N-terminus and far C-terminus. Chromosomal insertion mutations were generated in all seven genes. Null mutants could not be obtained for pmoC3, suggesting that it may play an essential role in growth on methane. Null mutants were obtained for pmoC1, pmoC2, pmoA1, pmoA2, pmoB1 and pmoB2. All of these mutants grew on methane, demonstrating that both gene copies were functional. Copy 1 mutants showed about two-thirds of the wild-type whole cell methane oxidation rate, while copy 2 mutants showed only about one-third of the wild-type rate, indicating that both gene copies were necessary for wild-type particulate methane monooxygenase activity. It was not possible to obtain double null mutants that were defective in both pmo copies, which may indicate that some expression of pMMO is important for growth. PMID- 10376841 TI - Expression of the Oenococcus oeni trxA gene is induced by hydrogen peroxide and heat shock. AB - Sequencing of the DNA region located upstream of the alpha-acetolactate synthase and decarboxylase (alsS-alsD) cluster of Oenococcus oeni allowed identification of an ORF, named trxA. This encodes a protein of 104 amino acids very similar to known thioredoxins. The protein encoded by the cloned fragment was able to complement Escherichia coli strains lacking a functional thioredoxin. Considering the results of protein sequence comparisons and complementation experiments, it was concluded that the trxA gene encodes a functional thioredoxin. Studies of trxA expression showed that the abundance of trxA mRNA was similar during all growth stages. A significant increase in trxA mRNA levels was observed in the presence of hydrogen peroxide in the medium or after heat shock. A single transcriptional start site was determined with total RNA isolated from cells subjected or not subjected to oxidative stress or heat shock. In each case the same promoter region was identified and shown to have a high similarity to the consensus promoter sequence of Gram-positive bacteria, as well as to that of E. coli and the previously mapped promoters from O. oeni. PMID- 10376842 TI - Structural and putative regulatory genes involved in cellulose synthesis in Rhizobium leguminosarum bv. trifolii. AB - Six genes involved in cellulose synthesis in Rhizobium leguminosarum bv. trifolii were identified using Tn5 mutagenesis. Four of them displayed homology to the previously cloned and sequenced Agrobacterium tumefaciens cellulose genes celA, celB, celC and celE. These genes are organized similarly in R. leguminosarum bv. trifolii. In addition, there were strong indications that two tandemly located genes, celR1 and celR2, probably organized as one operon, are involved in the regulation of cellulose synthesis. The deduced amino acid sequences of these genes displayed a high degree of similarity to the Caulobacter crescentus DivK and PleD proteins that belong to the family of two-component response regulators. This is to our knowledge the first report of genes involved in the regulation of cellulose synthesis. Results from attachment assays and electron microscopic studies indicated that cellulose synthesis in R. leguminosarum bv. trifolii is induced upon close contact with plant roots during the attachment process. PMID- 10376843 TI - Functional analysis of the O antigen glucosylation gene cluster of Shigella flexneri bacteriophage SfX. AB - Previous studies have shown that Shigella flexneri bacteriophage X (SfX) encodes a glucosyltransferase (GtrX, formerly Gtr), which is involved in O antigen modification (serotype Y to serotype X). However, GtrX alone can only mediate a partial conversion. More recently, a three-gene cluster has been identified next to the attachment site in the genome of two other S. flexneri bacteriophages (i.e. SfV and SfII). This gene cluster was postulated to be responsible for a full O antigen conversion. Here it is reported that besides the gtrX gene, the other two genes in the gtr locus of SfX were also involved in the O antigen modification process. The first gene in the cluster (gtrA) encodes a small highly hydrophobic protein which appears to be involved in the translocation of lipid linked glucose across the cytoplasmic membrane. The second gene in the cluster (gtrB) encodes an enzyme catalysing the transfer of the glucose residue from UDP glucose to a lipid carrier. The third gene (gtrX) encodes a bacteriophage specific glucosyltransferase which is largely responsible for the final step, i.e. attaching the glucosyl molecules onto the correct sugar residue of the O antigen repeating unit. A three-step model for the glucosylation of bacterial O antigen has been proposed. PMID- 10376844 TI - Molecular analysis of the recA gene and SOS box of the purple non-sulfur bacterium Rhodopseudomonas palustris no. 7. AB - The recA gene of the purple non-sulfur bacterium Rhodopseudomonas palustris no. 7 was isolated by a PCR-based method and sequenced. The complete nucleotide sequence consists of 1089 bp encoding a polypeptide of 363 amino acids which is most closely related to the RecA proteins from species of Rhizobiaceae and Rhodospirillaceae. A recA-deficient strain of R. palustris no. 7 was obtained by gene replacement. As expected, this strain exhibited increased sensitivity to DNA damaging agents. Transcriptional fusions of the recA promoter region to lacZ confirmed that the R. palustris no. 7 recA gene is inducible by DNA damage. Primer extension analysis of recA mRNA located the recA gene transcriptional start. A sequential deletion of the fusion plasmid was used to delimit the promoter region of the recA gene. A gel mobility shift assay demonstrated that a DNA-protein complex is formed at this promoter region. This DNA-protein complex was not formed when protein extracts from cells treated with DNA-damaging agents were used, indicating that the binding protein is a repressor. Comparison of the minimal R. palustris no. 7 recA promoter region with the recA promoter sequences from other alpha-Proteobacteria revealed the presence of the conserved sequence GAACA-N6-G(A/T)AC. Site-directed mutations that changed this consensus sequence abolished the DNA-damage-mediated expression of the R. palustris recA gene, confirming that this sequence is the SOS box of R. palustris and probably plays the same role in other alpha-Proteobacteria. PMID- 10376845 TI - Homocysteine, hypertension and stroke. AB - The balance of evidence from observational studies suggests that elevated homocysteine levels are associated with increased risk of carotid artery disease and stroke. There is however a paucity of prospective studies. There are also concerns regarding confounding due to factors associated with hyperhomocysteinemia, including renal impairment, an atherogenic diet and cigarette smoking. Homozygosity for a defective thermolabile variant of MTHFR, a common genetic polymorphism which results in hyperhomocysteinemia, has not been consistently linked with stroke or other vascular disease. There is a need for additional prospective studies with data on relevant confounders, sufficient power to characterise the form of the association between homocysteine concentrations and stroke risk, whether linear or threshold, and power to study interactions between homocysteine, other dietary markers and established stroke risk factors such as smoking and hypertension. Similarly, the evidence linking hyperhomocysteinemia with hypertension is limited and inconsistent. Given the biological mechanisms proposed in support of the homocysteine-CVD hypothesis, one would predict a positive association between homocysteine and blood pressure. There is a need to address this hypothesis directly in studies with reliable measurements of both homocysteine and blood pressure. Ultimately, the case for a causal role for elevated homocysteine levels in vascular disease, including hypertension and stroke, will depend on data from randomised controlled trials of homocysteine lowering interventions. Given the high prevalence of hyperhomocysteinemia in apparently well nourished populations and the tendency for homocysteine concentrations to increase with age, modest effects of homocysteine on stroke risk will have profound implications for public health. PMID- 10376847 TI - Reproducibility of ambulatory blood pressure monitoring in daily practice. AB - The reproducibility of ambulatory blood pressure monitoring (ABPM) was investigated in 45 untreated hypertensive patients in an out-patient clinic. Subjects with symptoms or diseases which could probably give rise to an increase in blood pressure (BP) variability were excluded. Patients underwent office BP (OBP) measurements and ABPM measurements with the Oxford Medilog device twice. The data were edited following previous set standards. Reproducibility of ABPM was good for the group: 24 h ABPM difference 0/2 mm Hg, standard deviation of the difference (SDD) 12/6 mm Hg for systolic BP and diastolic BP respectively. For OBP the difference between the two visits was 5/2 mm Hg with a SDD of 15/8 mm Hg. Intra-individual reproducibility was poor; almost half of the patients had a systolic difference of more than 10 mm Hg between both ABPM recordings. Reproduciblity of the day-night difference with a BP fall of at least 10% (dipper status) was moderate. About 60% of the subjects were dippers at one of the ABPM recordings but only 42% had a reproducible dip. Possible factors playing a role in the disappointing reproducibility of the ABPM recordings are the difference in daily activities between both recording days, decreased accuracy at higher BP, quality of sleep and the probable lower accuracy of the device during real ambulant conditions. In daily practice ABPM has no better reproducibility than OBP measurements, despite the larger number of measurements. PMID- 10376846 TI - Hypertension is associated with a high risk of cancer. AB - To investigate the possible relationship between hypertension and cancer, a retrospective analysis was carried out using a database including 1225 cases, of which 552 were hypertensives and 673 normotensives. Seventy cases of cancers with different origins were found during a 17-year follow-up. Odds ratio (OR) for occurrence of cancer was calculated. It was shown that an age over 40 years, male sex, alcohol-taking, systolic and diastolic blood pressures (SBP/DBP) were the five risk factors for the occurrence of cancers, while occupation, smoking, body mass index, left ventricular hypertrophy, and antihypertensive medication had no effect on cancer incidence. Hypertensives were at a high risk of overall cancer incidence with OR 2.2 (P < 0.01). After stratification of age, OR for hypertensives aged 40-49 years old with SBP > or =140 mm Hg or DBP > or =90mm Hg was 3.18 and 2.98 (P < 0.01 respectively). The OR of cancer for non-alcohol taking male hypertensives with SBP < or =140 mm Hg or DBP > or =90 mm Hg were 3.6 (95%CI 1.37-9.68, P = 0.003) and 5.67 (95%CI 2.01-16.75, P < 0.001), 7.55 (95%CI 2.10-33.19, P < 0.001) and 7.80 (95%CI 2.14-33.79, P < 0.001) for non-alcohol taking female hypertensives with SBP > or =140 mm Hg or DBP > or =90 mm Hg. After adjustment of age, sex and alcohol taking, the OR of the cancer incidence was 3.45 (95%CI 1.30-9.01, P < 0.01) for male and 5.0 (95%CI 1.56-16.67, P < 0.01) for female hypertensives aged 40-49 years. Multiple logistic regression analysis shows that age over 40 years, male sex, alcohol-taking, and DBP were the four independent risk factors for cancers. It is concluded that hypertension is associated with a high risk of cancer. PMID- 10376848 TI - Silent myocardial ischaemia in patients with essential arterial hypertension and non-insulin dependent diabetes mellitus. AB - The concomitant presence of diabetes mellitus and arterial hypertension significantly impairs myocardial function through a direct negative effect on cardiac myocytes, coronary microvessels and precipitation of atherosclerosis in major coronary arteries. The purpose of the present study was to establish to what extent non-insulin dependent diabetes mellitus (NIDDM) modified silent myocardial ischaemia (SMI) in patients with essential hypertension and without documented coronary artery disease (CAD). The study population consisted of 41 patients with essential arterial hypertension associated with NIDDM, treated with diet and oral hypoglycaemic agents (group I) and 40 patients with essential arterial hypertension without diabetes mellitus (group II). Both groups were comparable with respect to age, gender, duration, severity and complications of hypertension. A mean duration of diabetes mellitus in group I was 6.8 years. Conventional and automatic blood pressure and heart rate measurements, continuous ECG recordings, echocardiograms and laboratory tests were obtained in all patients. SMI was more frequent in group I than in group II (29.3% vs 12.5%, P < 0.05). In group I the total duration of SMI was longer (37.3 vs 2.8 min, P < 0.001) and the total number of silent episodes was larger (15.5 vs 2.6, P < 0.001). No inter-group differences were seen in conventional and automatic blood pressure and heart rate measurements. Both groups did not differ significantly in left ventricular mass index (LVMI) or the proportion of patients with left ventricular hypertrophy (LVH) (75.6% vs 60%). Lipid profile in both groups indicated an increased risk of CAD, but without significant differences. In conclusion, in patients with essential arterial hypertension and diabetes mellitus, the incidence and severity of SMI were clearly higher than in hypertensives with normal carbohydrate metabolism. Employment of modern diagnostic techniques in hypertensives permits identification of those at greater risk, which may have further clinical implications. PMID- 10376849 TI - Studies on cardiac sympathovagal balance and large artery distensibility in patients with untreated essential hypertension. AB - BACKGROUND: Power spectral analysis of heart rate variability and arterial distensibility are non-invasive measures of cardiac autonomic modulation and mechanical vessel wall properties, respectively. The aim of the present study was to assess cardiac sympathovagal balance, carotid and brachial artery distensibility and a possible relation between these parameters in mildly hypertensive patients as compared to normotensive controls. METHODS: Total power (TP, 0.01 to 0.5 Hz) and spectral components (low frequency 0.04-0.15 Hz, mainly sympathetic cardiac modulation; high frequency 0.15-0.4 Hz, mainly vagal cardiac modulation) and cardiac sympathovagal balance (LF/LH ratio) of short term heart rate variability (ECG-recording) were calculated in 15 untreated essential hypertensive patients (HYP) and 15 age- and sex-matched healthy controls (CON). Brachial and carotid artery distensibility coefficient (DC) was measured with a multigate doppler system (echo-tracking). RESULTS: TP (ms2 x 10(-3)) (11.2 +/- 0.8 vs 13.6 +/- 0.9, P < 0.03), LF/HF ratio (1.07 +/- 0.08 vs 0.75 +/- 0.07, P < 0.01) and HF (ms2 x 10(-3)/%) (0.7 +/- 0.1/49 +/- 2 vs 1.3 +/- 0.2/58 +/- 2, P < 0.01/P < 0.01) were significantly reduced in HYP compared to CON subjects. LF (ms2 x 10(-3)/%) was 0.7 +/- 0.1/50 +/- 2 vs 0.9 +/- 0.1/41 +/- 2, P = 0.16/P < 0.01. Carotid artery DC (15 +/- 2 vs 26 +/- 2, P < 0.001) and brachial artery DC (4.7 +/- 0.6 vs 9 +/- 1.0, P < 0.001) were significantly reduced in HYP. There was a significant correlation between carotid DC and LF/HF (rho = -0.41, P < 0.03). CONCLUSION: The data shows reduced heart rate variability and altered cardiac sympathovagal balance as well as impaired arterial distensibility in untreated mildly hypertensive patients. The relative increase in sympathetic modulation and decreased carotid distensibility appear to be related. PMID- 10376850 TI - Effect of protein kinase C and insulin on Na+/H+ exchange in red blood cells of essential hypertensives. AB - The kinetic properties of sodium-proton exchange are abnormal in human red blood cells of hypertensive patients and it has been demonstrated that the transport protein undergoes post-translational modifications able to affect its kinetic properties. Protein kinase C (PKC) activation decreases the affinity constant for intracellular protons while insulin increases the maximal rate of proton translocation. The present study therefore aimed to examine the relationships among PKC activity, fasting insulin levels and the kinetic behaviour of sodium proton exchange in red blood cells from 20 normotensives and 36 hypertensives. In comparison with normotensive subjects, hypertensive patients had higher body mass index (26.2 +/- 0.7 vs 23.6 +/- 0.6 kg/m2, P < 0.05), higher fasting insulin levels (93.2 +/- 10.8 vs 38.6 +/- 2.9 pmol/L), increased maximal velocity of proton translocation (37.9 +/- 2.7 vs 27.6 +/- 1.9 mmol/L per cell x h, P < 0.05), and reduced Hill's coefficient (1.6 +/- 0.1 vs 2.0 +/- 0.1, P < 0.01) of sodium-proton exchange. Basal PKC activity of the cytosol and membrane was similar in the study groups. However, after treatment with 1 micromol/L phorbol 12-myristate 13-acetate (PMA) for 10 min, membrane PKC activity was stimulated to a larger extent in hypertensives (to 181 +/- 8 pmol/min/mg protein) than in normotensives (to 136 +/- 6 pmol/min/mg protein, P < 0.01). The PMA stimulated PKC activity was positively correlated to fasting insulin levels (r = 0.59, P < 0.01). Stimulation of membrane PKC by PMA corrected the low Hill's coefficient for H(i)+ activation of sodium-proton exchange in the hypertensives, while the constant for half maximal activation for intracellular protons (ie, the affinity for intracellular protons) decreased to a similar extent in both groups. The maximal transport rate was unaffected by PMA. These results indicate that the abnormal proton activation of red blood cell sodium-proton exchange in hypertensives reflects an abnormal regulation of PKC translocation to the cell membrane, associated to hyperinsulinaemia and probably insulin resistance. Therefore, post-translational modifications of the transport protein(s) account for the altered kinetic behaviour of sodium-proton exchange in hypertensives. PMID- 10376851 TI - Plasma insulin, plasminogen activator inhibitor, and ankle-brachial systolic blood pressure ratio in overweight hypertensive subjects. AB - BACKGROUND: In hypertensive subjects, the ratio between ankle and brachial systolic blood pressure (ABI) has been shown to be an independent risk factor for cardiovascular diseases, particularly in the elderly. Plasma insulin may be an important interconnecting factor explaining this observation. PURPOSE: In a population of middle-aged subjects with essential hypertension and moderate overweight, we identified whether the decrease in the ABI ratio was associated with the clinical and biochemical factors involved in resistance to insulin. Patients with diabetes and/or arteriosclerosis obliterans of the lower limbs were excluded from the population. Subjects were or were not on antihypertensive therapy. RESULT: On the basis of univariate correlations, the ABI ratio was found to be significantly and negatively associated not only with the degree of abdominal fat distribution, but also with the usual biological features of resistance to insulin: plasma triglycerides and cholesterol; plasma glucose and insulin; and plasminogen activator inhibitor (PAI) antigen. In a multivariate analysis in subjects with untreated hypertension, the ABI ratio was significantly and negatively associated with only three variables: age, plasma insulin and PAI antigen. In treated hypertensive subjects, only the role of age and insulin remained significant. CONCLUSION: Since the alterations of the ABI ratio may be considered as a marker of the changes in the structure and function of arteries of the lower limbs, the study provides evidence that plasma insulin and PAI antigen, independently of the presence of significant atherosclerotic occlusive lesions, are susceptible to alter the pressure wave transmission in conduit arteries of the lower limbs. PMID- 10376852 TI - Co-administration of an ACE-inhibitor (moexipril) and hormonal replacement therapy in postmenopausal women. AB - Co-administration of antihypertensive drug therapy and hormonal replacement therapy (HRT) is frequent in postmenopausal women but it is not known whether HRT interacts with concomitant antihypertensive therapy. The present study was designed to investigate efficacy and safety of the ACE inhibitor moexipril in comparison to placebo in hypertensive, postmenopausal women on HRT. After a 4 week placebo run-in phase, 95 postmenopausal women (35-74 years of age) who had a sitting diastolic blood pressure (BP) of 95-114 mm Hg and were treated with HRT were randomised to a 12-week treatment with moexipril 15 mg or placebo. Efficacy and safety were assessed by measuring changes in sitting BP and metabolic parameters associated with cardiovascular disease including triglycerides, total cholesterol, HDL, LDL, total cholesterol/HDL ratio and glucose. Adverse events were recorded continuously. After 12 weeks of treatment, moexipril 15 mg was significantly more effective in reducing sitting systolic and diastolic BP from baseline than placebo (-12.2/-9.9 mm Hg vs -1.6/-4.3 mm Hg, P < 0.001). Metabolic parameters were not affected by treatment with moexipril: mean levels of triglycerides, total cholesterol, HDL, LDL, total cholesterol/HDL ratio and glucose remained unchanged throughout the study. Fibrinogen, an independent cardiovascular risk factor, increased after placebo (+35.0 mg/dl) and decreased after treatment with moexipril (-33.6 mg/dl), the difference, however, was not statistically significant. Moexipril was well-tolerated by postmenopausal women using HRT. The most frequent adverse events included headache (21.3%), cough (12.8%) and rhinitis (10.6%) and there were no significant differences in the number and severity of adverse events between the moexipril and placebo groups. This study indicates that moexipril is effective and well tolerated in the treatment of hypertensive, postmenopausal women and can safely be co-administered to HRT. PMID- 10376853 TI - A man with severe, transient hypertension due to acute renal infarction. PMID- 10376854 TI - The St George's 'Star' and 'Imploding Diamond'. PMID- 10376855 TI - Cervical cancer, Pap smear and HPV testing: an update of the role of organized Pap smear screening and HPV testing. PMID- 10376856 TI - The Icelandic and Nordic cervical screening programs: trends in incidence and mortality rates through 1995. AB - BACKGROUND: The objective of cervical cancer screening is to lower the incidence and mortality rates of the disease. This study evaluates the effectiveness of cervical screening and the UICC and EC screening recommendations based on the Nordic screening experience. METHODS: The study analyzes the features of the Icelandic and the Nordic screening programs and the observed trends in the incidence and mortality rates in these countries through 1995. RESULTS: Organized screening started in all the Nordic countries soon after 1960 and had nation-wide coverage in all these countries, except in Denmark (45% coverage in 1991), by around 1973 but in Norway screening was only spontaneous up to late in 1994. Up to 1985 the target age group and screening interval were most intensive in Iceland, followed by Finland, Sweden and Denmark. All countries except Finland lowered the lower age limit and intensified the screening intervals after 1985. Through the period 1986-1995 the reduction in both the mortality and the incidence rates was greatest in Iceland (mortality: 76% and incidence: 67%) and Finland (73% and 75%, respectively), intermediate in Sweden (60% and 55%, respectively) and Denmark (55% and 54%, respectively), and lowest in Norway (43% and 34%, respectively). The age-specific incidence in the 20-29 age group has been increasing since 1971 in all the Nordic countries, except in Finland, where the yearly registered age-specific incidence has been increasing in the targeted 30-54 age group since 1991. In Iceland screening has greatly affected the rate of all stages of squamous cell carcinoma, but not the rate of adeno- and adenosquamous carcinomas. In fact the rate of adenocarcinoma has been increasing. CONCLUSIONS: Organized screening is more effective than spontaneous screening in reducing the risk of cervical cancer. Although differences in environmental, biological and ethnic factors may call for different screening strategies, screening should preferably start soon after age 20 with a screening interval of 2-3 years. PMID- 10376857 TI - Trends in cervical intra-epithelial neoplasia in Iceland through 1995: evaluation of targeted age groups and screening intervals. AB - BACKGROUND: Targeted age groups and screening intervals are dependent on the age specific prevalence of the preclinical disease and the length of the detectable pre-clinical phase. This study evaluates the UICC and EC recommendations regarding targeted age group and screening intervals based on the Icelandic screening experience. METHODS: The trends for cytologic preinvasive lesions were analyzed at first visit during the period 1966-1995, at second and later visits after a normal test(s) taken after 1985, and finally at any visit in 1991-1995. The frequency of histologic lesions was calculated for the birth cohort from the age of 60 and among women referred for colposcopic examination in 1994. RESULTS: The prevalence of preinvasive disease has increased significantly since 1980, and the rate of moderate to high-grade cytologic and histologic lesions begins increasing as early as 20 years of age. The rate of these lesions starts to accumulate at 24 to 36 months after a normal smear, but the rate decreases with the number of negative smears taken. Among correctly screened women the rate of histologically verified moderate to high-grade lesions and invasive disease is practically non-existent after the age of 60, while among the younger women cases with microinvasive disease start to appear within 2 to 3 years after a normal smear. CONCLUSION: Screening should start soon after age 20 with a screening interval of 2 to 3 years. The screening interval can probably be extended to 4 years at the age of 50 and stop at the age of 60 to 64 among regularly screened women. PMID- 10376858 TI - The effect of betamethasone and dexamethasone on fetal heart rate patterns and biophysical activities. A prospective randomized trial. AB - BACKGROUND; Contradictory findings on the effect of betamethasone versus dexamethasone on antenatal tests of fetal well-being have been reported. The purpose of this study was to compare the effects of these steroid compounds on fetal heart rate patterns and biophysical activities in a prospective. randomized trial. STUDY DESIGN: Forty-six pregnant women (gestational age range 27-34 weeks) at risk for preterm delivery were randomized to receive betamethasone or dexamethasone for enhancement of fetal lung maturity. Fetal heart rate was recorded for 60 minutes and analyzed with the Sonicaid System 8000 before (0 hours), and 48 hours and 96 hours after steroid administration. Subsequently, fetal limb, body and breathing movements were sonographically observed and quantified for 30 minutes. To account for fetal circadian rhythms, all examinations were performed between 1 p.m. and 5 p.m., at least one hour after maternal meals. RESULTS: Fetal heart rate accelerations (p<0.001; p<0.01), short term variation (p<0.0001; p<0.05), long-term variation (p<0.01; p=NS), duration of high episodes (p<0.001; p<0.05), total movement count (p<0.001; p<0.05), and duration of breathing time (p<0.0001; p<0.0001) were substantially reduced 48 h after betamethasone and dexamethasone administration, respectively, with percent reduction being larger for the betamethasone group, except for breathing movements (p<0.05; p<0.001; p<0.001; p<0.005; p<0.05; p=NS; respectively). In 68.2%( and 45.5% of fetuses, less than 30 seconds of continuous breathing movements were found in the betamethasone and dexamethasone groups, respectively. In 71.8% and 12.5%, of fetuses, respectively, less than 2 body/limb movements were observed. Therefore five and two fetuses in the betamethasone and dexamethasone study group, respectively, had both nonreactive fetal heart rate monitors for 60 minutes and biophysical profiles of < or =4/10. All parameters returned to baseline values at 96 h. Baseline fetal heart rate and numbers of decelerations remained unchanged (p=NS). CONCLUSIONS: Both betamethasone and dexamethasone induce a profound, albeit transient, suppression of fetal heart rate characteristics and biophysical activities in the preterm fetus. However, the effect of betamethasone is more pronounced. Awareness of these phenomena might prevent unwarranted iatrogenic delivery of preterm fetuses. PMID- 10376859 TI - MULTISCAN--a Scandinavian multicenter second trimester obstetric ultrasound and serum screening study. AB - AIM: To study the detection rates of second trimester ultrasound screening for neural tube defects (NTD), abdominal wall defects (AWD) and Down's syndrome (DS) in low risk populations at tertiary centers, and to compare the ultrasound screening detection rates with those that were obtainable by biochemical serum screening (double test: alpha-fetoprotein/human chorion gonadotrophin/age test). STUDY DESIGN: Prospective multicenter study with a three year inclusion period: 1/1/1989-31/12/1991. SUBJECTS: 27,844 low-risk women at 18-34 years of age who had a second trimester ultrasound screening examination. Of these, 10,264 also had a serum test. METHODS: An ultrasound malformation scan and a serum test were carried out at 17-19 weeks of gestation. Risk calculations regarding DS were based on alpha-fetoprotein, human chorion gonadotrophin and maternal age; performed retrospectively for the first two years. RESULTS: In total 73 cases were identified in the study population: NTD (n=34), AWD (n=7) and DS (n=32). The detection rates, (%, with 95% confidence interval) for ultrasound screening were: NTD: 79.4 (62.1-91.3); AWD: 85.7 (42.1-99.6); DS: 6.3 (0.8-20.8). In the subgroup of women who had both tests, the detection rates for ultrasound screening vs double test were: NTD: 62.5 (24.5-91.5) vs 75.0 (34.9-96.8); AWD: 66.7 (9.4-99.2) vs 100 (29.2-100.0); DS: 7.7 (0.2-36.0) vs 46.2 (19.2-74.9). The false positive rates (%) for ultrasound screening vs double test were: NTD: 0.01/3.3; AWD: 0.01/3.3; DS: 0.1/4.0. CONCLUSION: Second trimester ultrasound screening in a low risk population gave a low detection rate for fetal DS (6.3%) and an acceptable detection rate for NTD (79.4%) and AWD (85.7%). In the subgroup of women who had both tests, serum screening performed better than ultrasound as applied in the present study, especially regarding DS. PMID- 10376860 TI - Induced second trimester abortion by extra-amniotic prostaglandin infusion in patients with a cesarean scar: is it safe? AB - BACKGROUND: One result of the advancement in prenatal diagnosis is an increase in the need for second trimester pregnancy terminations. Extra-amniotic infusion of prostaglandins is a common technique used for such pregnancy termination. Since prostaglandins cause strong uterine contractions, many practitioners are hesitant to use this technique on women with a uterine scar. In this study we tried to evaluate the effectiveness and safety of the technique for women with a previous uterine scar. METHODS: This retrospective study included all women with a complete medical record who underwent a second trimester pregnancy termination at our institution by extra amniotic prostaglandin E2, during a 6 year period. The study group included all women with a previous uterine scar. The group of women without such a scar served as the control group. RESULTS: Three hundred and forty women had their pregnancy terminated, but only in 282 cases was the medical information complete (research population). The study group (35 women) characteristics were similar to those of the control group (247 women). We found no difference in the abortion interval, the need to use an additional method, the need for curettage and in bleeding complication between the two groups. There was no case of uterine rupture. The group of women with multiple uterine scars was too small for analysis. CONCLUSIONS: Our results suggest that extra amniotic prostaglandin infusion is an effective and safe technique in women with a uterine scar. PMID- 10376861 TI - Effect of L-arginine load on platelet aggregation: a comparison between normotensive and preeclamptic pregnant women. AB - BACKGROUND: In the present study we hypothesized that a derangement of the L arginine-nitric oxide system could be involved in the development of the hypercoagulative status found during preeclampsia. In order to verify such hypothesis we have compared the effects of nitric oxide substrate, L-arginine on platelet aggregation. Moreover, we have also measured the L-citrulline plasma levels as a stochiometric metabolite resulting from the conversion L-arginine to nitric oxide. METHODS: Nine preeclamptic women and 11 normotensive pregnant women were enrolled for the study. Subjects were infused with saline and with 30gr of L arginine. Blood samples were drawn during the saline infusion (30 min), during L arginine administration (30 min) and 30 min thereafter. ADP and collagen-induced platelet aggregation was studied as per Born with a dual-channel aggregometer (Chrono-Log, Mascia Brunelli, Italy) and L-citrulline was measured by HPLC. RESULTS: In normotensive women the infusion significantly decreased ADP and collagen-induced aggregation after 15 minutes of L-arginine load; whereas no effects were observed in preeclamptic women. Similarly in normotensive but not in preeclamptic women L-arginine load was able to increase L-citrulline plasma levels. CONCLUSIONS: In normotensive women the in vivo L-arginine administration decreases platelet aggregation with an increase of L-citrulline plasma levels. On the contrary, no effects were observed in preeclamptic women. These findings confirm that a hypercoagulative status characterizes preeclampsia and that such phenomenon could be explained by a derangement of the platelet L-arginine-nitric oxide pathway. PMID- 10376862 TI - Birthweight in women with potential gestational diabetes mellitus--an effect of obesity rather than glucose intolerance? AB - BACKGROUND: The purpose was to compare the influence of varying levels of glycemia on the perinatal outcome. METHODS: The data charts of 383 women screened for gestational diabetes mellitus with an oral glucose tolerance test during two birthyears were retrospectively evaluated. In 55 women gestational diabetes mellitus was diagnosed and treated with diet. The non-diabetic women (n=328) were subdivided into a borderline diabetes group (n=74) and a normal group (n= 254) on the basis of the oral glucose tolerance test result. The birth registry of 8196 singleton pregnancies from The Perinatal Research Unit at Skejby University Hospital served as the background population. RESULTS: Birthweight was highest in the borderline group. Weight increase during pregnancy was larger in the non diabetic than the gestational diabetic women (15 vs. 8 kg p<0.01). The women with less increase of body weight delivered neonates with lower birthweight than those with higher increase. Birthweight was associated with maternal weight during pregnancy (p<0.01). Birthweight ratio increased with increasing glucose intolerance. Vaginal delivery rate was less and cesarean section rate higher in women with gestational diabetes mellitus compared to the non-diabetic women. No significant difference was found in the incidence of hypertensive disorders during pregnancy or neonatal morbidity. CONCLUSIONS: Even minor hyperglycemia is associated with increasing birthweight. Birthweight is reduced in GDM when dietary treatment is instituted and effect on weight gain is achieved. PMID- 10376863 TI - Sperm morphology and IVF: embryo quality in relation to sperm morphology following the WHO and Kruger's strict criteria. AB - BACKGROUND: The aim of this study was to examine the correlation between sperm morphology and embryo quality/IVF outcome. METHODS: The implication of sperm morphology assessment before an IVF cycle was evaluated. A total of 100 IVF couples where the female partner had either tubal factor (n=50) or unexplained infertility (n= 50) entered a prospective study, and sperm samples for the actual cycle were assessed according to the strict criteria and WHO criteria. The study was blinded for the technician involved in sperm morphology analyzing. IVF was carried out according to a long down regulation protocol using GnRH/FSH/hCG and ova were inseminated with 200,000 spermatozoa/ml. Embryos were transferred on day 2 post fertilization in a maximum of three embryos. RESULTS: No significant differences were found between the groups regarding age of the female partner (mean=34.3), no. oocytes retrieved (mean=8.5), fertilization (66.5%), pregnancies (pos. S-hCG/transfer 39.6%) or 'Take home baby rate' (birth rate/transfer 30.0%). As to the score of Kruger's strict criteria and the WHO criteria, we found no correlation between this score and cleavage rate, embryo development or pregnancies. The WHO criteria were found to be a better predictor for fertilization rate than the Kruger's criteria (p<0.002). CONCLUSION: The strict criteria or sperm evaluation according to WHO have no better predictive value for the outcome of routine IVF. PMID- 10376864 TI - Comparison of ovarian electrocautery and oral contraceptives in the treatment of hyperandrogenism in women with polycystic ovary syndrome. AB - BACKGROUND: Endocrine treatment of hyperandrogenism in women with polycystic ovary syndrome (PCOS) aims at reduction of androgens and increasing sex hormone binding globuline (SHBG), which are also side effects of ovarian electrocautery (OE) when used for induction of ovulation. METHODS: Hormonal effects of ovarian electrocautery were compared with the effects of oral contraceptives (OC) containing desogestrel (DG) or cyproteron acetate (CPA). OCs were given to 18 women with PCOS as their sole treatment (group A) and to 23 women after the restoration of regular ovulatory cycling by ovarian electrocautery (group B). RESULTS: Ovarian electrocautery induced ovulation and increased the concentration of estrogens and especially progesterone, while OC induced the opposite effects. In the androgens and SHBG the two treatments induced changes that were in the same direction, but OC treatment induced changes that exceeded those of OE. The concentration of SHBG increased from 27.9 to 127.7 nmol/L on OC treatment (Group A), compared with 37.2 to 44.9 after OE (Group B). The androgens decreased, for testosterone the decreases were 2.1 and 0.99 nmol/L, respectively, for androsterone 5.36 and 3 nmol/L, for dihydrotestosterone 0.12 and 0.1 nmol/L, and for DHEAS 3.28 and 1.8 umol/L. No further gain was obtained by the combination of the two treatments. CONCLUSIONS: Hyperandrogenism in women with PCOS can be effectively treated with OCs containing DG or CPA. In women with concurrent infertility, however, ovarian electrocautery can be recommended. The only indication for the combination of these two treatments is ambivalence with regard to the fertility after ovulation induction by OE. A temporary delay of ovulatory cycling with OC-treatment after OE has no negative impact upon the future fertility. PMID- 10376865 TI - Compliance to estrogen treatment one to three years after hysterectomy and bilateral salpingooophorectomy. The cohort's lifestyle, knowledge of ERT, benefits etc. AB - BACKGROUND: To determine compliance to Estrogen Replacement Therapy (ERT) initiated at discharge from a gynecological department in perimenopausal women having had hysterectomy (hyst.) and bilateral salpingooophorectomy (BSO) for benign disease. To investigate this cohort's knowledge and attitude to ERT, lifestyle issues, sources of information etc. METHODS: A questionnaire was sent to the patients 1-3 years after discharge (April 1993 to December 1994). RESULTS: Eighty-eight patients were identified and returned the questionnaire. Seventy nine were included in the statistics. Eighty-nine percent were still compliant to treatment. Forty-two percent received their information about ERT from newspapers/magazines and 17%) from GPs. Ninety percent knew that ERT prevented osteoporosis, 33% knew about the positive effect on cardiovascular disease (CVD). Sixty-nine percent believed that ERT increased the risk of breast cancer and 74% anticipated an increase in quality of life. CONCLUSION: Even though not offered any follow-up visits, 89% of this cohort was still on ERT 1-3 years after hyst. and BSO. This high rate of compliance might be due to initiation of treatment during hospitalization with the information given at discharge combined with a low rate of side effects of ERT. The women's primary source of information was newspapers/ magazines, and they were aware of the benefits of ERT on climacteric symptoms and osteoporosis but not so much the benefits on CVD. The major concerns were the risk of breast cancer and weight gain. PMID- 10376866 TI - Insulin sensitivity during postmenopausal hormone replacement with transdermal estradiol and intrauterine levonorgestrel. AB - BACKGROUND: The study was devised to measure the effect of intrauterinely delivered levonorgestrel and transdermal estradiol on insulin sensitivity in postmenopausal women and compare this effect with that induced by transdermal estradiol alone. METHODS: An open, prospective, comparative study of healthy postmenopausal women without earlier use of hormone replacement therapy. The estrogen therapy group consisted of eight hysterectomized women, who used a transdermal patch delivering a daily dose of 50 microg of estradiol continuously for 6 months. The estrogen-progestin therapy group consisted of 13 women with an intact uterus, who received a simultaneous combination of a transdermal patch and a levonorgestrel (20 microg/day) intrauterine system for the same length of time. Fasting plasma concentrations of glucose, insulin and C-peptide and an insulin tolerance test were used to measure glucose metabolism and insulin sensitivity. RESULTS: Neither therapy changed the fasting plasma levels of glucose, insulin or C-peptide. Transdermal estrogen improved insulin sensitivity by 22%, as measured by an insulin tolerance test, while a small increase of 3.6% was observed using the combination therapy. CONCLUSIONS: Transdermal estradiol improves insulin sensitivity in healthy postmenopausal women. Combining intrauterine levonorgestrel to transdermal estradiol reverses this effect. This combination does not, however, seem to induce insulin resistance. PMID- 10376867 TI - The prevalence of urinary incontinence and its influence on the quality of life in women from an urban Swedish population. AB - OBJECTIVES: To assess the prevalence of urinary incontinence and its influence on the quality of life. MATERIAL AND METHODS: A random sample of every fourth woman aged > or =20 years resident in a primary health care district of the city of Goteborg was obtained from the population register (n=2911). The women were invited by letter to complete a questionnaire concerning urinary incontinence. The women were also requested to assess their quality of life using a visual analogue scale. RESULTS: The overall response rate was 77%. The prevalence of urinary incontinence increased (p<0.001) in a linear fashion from 3% in the cohort 20-29 years to 32 % in the cohort of women aged > or =80 years. The proportion of women suffering from stress incontinence decreased (p<0.001) with increasing age, while the proportion of women suffering from urge and mixed incontinence increased (p<0.01) with increasing age. Women with stress incontinence had a greater body weight and had given birth to a greater number of children compared to continent women. There was, however, in this respect no difference between women with urge incontinence and continent women. Women with urinary incontinence reported a poorer quality of life compared to continent women (p<0.01). Women with urge incontinence and women with mixed incontinence reported a poorer quality of life compared to women with stress incontinence (p<0.05). Only 6% of the women from this population had sought medical attention for urinary incontinence. CONCLUSIONS: Although urinary incontinence was a prevalent condition, particularly among the elderly and had a negative influence on the quality of life, only a small number of women had sought medical care. PMID- 10376868 TI - Safety clamp and cutter (SCC 23) for the umbilical cord. PMID- 10376869 TI - Cerebral hyperperfusion in patient with eclampsia. PMID- 10376870 TI - Is the use of IGFB-1 for diagnosing ROM of any clinical value. PMID- 10376872 TI - A nested double pseudoknot is required for self-cleavage activity of both the genomic and antigenomic hepatitis delta virus ribozymes. AB - The crystal structure of a genomic hepatitis delta virus (HDV) ribozyme 3' cleavage product predicts the existence of a 2 bp duplex, P1.1, that had not been previously identified in the HDV ribozymes. P1.1 consists of two canonical C-G base pairs stacked beneath the G.U wobble pair at the cleavage site and would appear to pull together critical structural elements of the ribozyme. P1.1 is the second stem of a second pseudoknot in the ribozyme, making the overall fold of the ribozyme a nested double pseudoknot. Sequence comparison suggests the potential for P1.1 and a similar fold in the antigenomic ribozyme. In this study, the base pairing requirements of P1.1 for cleavage activity were tested in both the genomic and antigenomic HDV ribozymes by mutagenesis. In both sequences, cleavage activity was severely reduced when mismatches were introduced into P1.1, but restored when alternative base pairing combinations were incorporated. Thus, P1.1 is an essential structural element required for cleavage of both the genomic and antigenomic HDV ribozymes and the model for the antigenomic ribozyme secondary structure should also be modified to include P1.1. PMID- 10376871 TI - mRNA surveillance in eukaryotes: kinetic proofreading of proper translation termination as assessed by mRNP domain organization? AB - In the last few years it has become clear that a conserved mRNA degradation system, referred to as mRNA surveillance, exists in eukaryotic cells to degrade aberrant mRNAs. This process plays an important role in checking that mRNAs have been properly synthesized and functions, at least in part, to increase the fidelity of gene expression by degrading aberrant mRNAs that, if translated, would produce truncated proteins. A critical issue is how normal and aberrant mRNAs are distinguished and how that distinction leads to differences in mRNA stability. Recent results suggest a model with three main points. First, mRNPs have a domain organization that is, in part, a reflection of the completion of nuclear pre-mRNA processing events. Second, the critical aspect of distinguishing a normal from an aberrant mRNA is the environment of the translation termination codon as determined by the organization of the mRNP domains. Third, the cell distinguishes proper from improper termination through an internal clock that is the rate of ATP hydrolysis by Upf1p. If termination is completed before ATP hydrolysis, the mRNA is protected from mRNA degradation. Conversely, if termination is slow, then ATP hydrolysis and a structural rearrangement occurs before termination is completed, which affects the fate of the terminating ribosome in a manner that fails to stabilize the mRNA. This proposed system of distinguishing normal from aberrant transcripts is similar to, but distinct from other systems of kinetic proofreading that affect the accuracy of other biogenic processes such as translation accuracy and spliceosome assembly. PMID- 10376873 TI - A potential mechanism for selective control of cap-independent translation by a viral RNA sequence in cis and in trans. AB - Highly efficient cap-independent translation initiation at the 5'-proximal AUG is facilitated by the 3' translation enhancer sequence (3'TE) located near the 3' end of barley yellow dwarf virus (BYDV) genomic RNA. The role of the 3'TE in regulating viral translation was examined. The 3'TE is required for translation and thus replication of the genomic RNA that lacks a 5' cap (Allen et al., 1999, Virology253:139-144). Here we show that the 3'TE also mediates translation of uncapped viral subgenomic mRNAs (sgRNA1 and sgRNA2). A 109-nt viral sequence is sufficient for 3'TE activity in vitro, but additional viral sequence is necessary for cap-independent translation in vivo. The 5' extremity of the sequence required in the 3' untranslated region (UTR) for cap-independent translation in vivo coincides with the 5' end of sgRNA2. Thus, sgRNA2 has the 3'TE in its 5' UTR. Competition studies using physiological ratios of viral RNAs showed that, in trans, the 109-nt 3'TE alone, or in the context of 869-nt sgRNA2, inhibited translation of genomic RNA much more than it inhibited translation of sgRNA1. The divergent 5' UTRs of genomic RNA and sgRNA1 contribute to this differential susceptibility to inhibition. We propose that sgRNA2 serves as a novel regulatory RNA to carry out the switch from early to late gene expression. Thus, this new mechanism for temporal control of translation control involves a sequence that stimulates translation in cis and acts in trans to selectively inhibit translation of viral mRNA. PMID- 10376874 TI - C-terminal interaction of translational release factors eRF1 and eRF3 of fission yeast: G-domain uncoupled binding and the role of conserved amino acids. AB - Translation termination in eukaryotes requires a stop codon-responsive (class-I) release factor, eRF1, and a guanine nucleotide-responsive (class-II) release factor, eRF3. Schizosaccharomyces pombe eRF3 has an N-terminal polypeptide similar in size to the prion-like domain of Saccharomyces cerevisiae eRF3 in addition to the EF-1alpha-like catalytic domain. By in vivo two-hybrid assay as well as by an in vitro pull-down analysis using purified proteins of S. pombe as well as of S. cerevisiae, eRF1 bound to the C-terminal one-third domain of eRF3, named eRF3C, but not to the N-terminal two-thirds, which was inconsistent with the previous report by Paushkin et al. (1997, Mol Cell Biol 17:2798-2805). The activity of S. pombe eRF3 in eRF1 binding was affected by Ala substitutions for the C-terminal residues conserved not only in eRF3s but also in elongation factors EF-Tu and EF-1alpha. These single mutational defects in the eRF1-eRF3 interaction became evident when either truncated protein eRF3C or C-terminally altered eRF1 proteins were used for the authentic protein, providing further support for the presence of a C-terminal interaction. Given that eRF3 is an EF Tu/EF-1alpha homolog required for translation termination, the apparent dispensability of the N-terminal domain of eRF3 for binding to eRF1 is in contrast to importance, direct or indirect, in EF-Tu/EF-1alpha for binding to aminoacyl-tRNA, although both eRF3 and EF-Tu/EF-1alpha share some common amino acids for binding to eRF1 and aminoacyl-tRNA, respectively. These differences probably reflect the independence of eRF1 binding in relation to the G-domain function of eRF3 (i.e., probably uncoupled with GTP hydrolysis), whereas aminoacyl-tRNA binding depends on that of EF-Tu/EF-1alpha(i.e., coupled with GTP hydrolysis), which sheds some light on the mechanism of eRF3 function. PMID- 10376875 TI - 16S ribosomal RNA pseudouridine synthase RsuA of Escherichia coli: deletion, mutation of the conserved Asp102 residue, and sequence comparison among all other pseudouridine synthases. AB - The gene for RsuA, the pseudouridine synthase that converts U516 to pseudouridine in 16S ribosomal RNA of Escherichia coli, has been deleted in strains MG1655 and BL21/DE3. Deletion of this gene resulted in the specific loss of pseudouridine516 in both cell lines, and replacement of the gene in trans on a plasmid restored the pseudouridine. Therefore, rsuA is the only gene in E. coli with the ability to produce a protein capable of forming pseudouridine516. There was no effect on the growth rate of rsuA- MG1655 either in rich or minimal medium at either 24, 37, or 42 degrees C. Plasmid rescue of the BL21/DE3 rsuA- strain using pET15b containing an rsuA gene with aspartate102 replaced by asparagine or threonine demonstrated that neither mutant was active in vivo. This result supports a role for this aspartate, located in a unique GRLD sequence in this gene, at the catalytic center of the synthase. Induction of wild-type and the two mutant synthases in strain BL21/DE3 from genes in pET15b yielded a strong overexpression of all three proteins in approximately equal amounts showing that the mutations did not affect production of the protein in vivo and thus that the lack of activity was not due to a failure to produce a gene product. Aspartate102 is found in a conserved motif present in many pseudouridine synthases. The conservation and distribution of this motif in nature was assessed. PMID- 10376876 TI - The properties of chimeric picornavirus IRESes show that discrimination between internal translation initiation sites is influenced by the identity of the IRES and not just the context of the AUG codon. AB - The internal ribosome entry segment (IRES) of picornaviruses consists of approximately 450 nt of 5'-untranslated region, terminating at the 3' end with an approximately 25 nt element consisting of an absolutely conserved UUUC motif followed by a more variable pyrimidine-rich tract and G-poor spacer, and finally an AUG triplet, which is considered to be the actual ribosome entry site. Events following entry at this site differ among picornaviruses: in encephalomyocarditis virus (EMCV) virtually all ribosomes initiate translation at this site (AUG-11); in foot-and-mouth-disease virus (FMDV), one-third of the ribosomes initiate at this AUG (the Lab site), and the rest at the next AUG 84 nt downstream (Lb site); and in poliovirus (PV), the AUG at the 3' end of the IRES (at nt 586 in PV type 1) is considered to be a silent entry site, with all ribosomes initiating translation at the next AUG downstream (nt 743). To investigate what determines this different behavior, chimeras were constructed with a crossover at the conserved UUUC motif: the body of the IRES, the sequences upstream of this UUUC motif, was derived from one species, and the downstream sequences from another. When the body of the FMDV or PV IRESes was replaced by that of EMCV, there was a marked increase in the absolute and relative frequency of initiation at the upstream AUG, the Lab site of FMDV and 586AUG of PV, respectively. In contrast, when the body of the EMCV IRES was replaced by that of PV, initiation occurred with no preference at three AUGs: the normal site (AUG-11), AUG-10 situated 8 nt upstream, and AUG-12, which is 12 nt downstream. Thus although the context of the AUG at the 3' end of the IRES may influence initiation frequency at this site, as was shown by improving the context of 586AUG of PV, the behavior of the ribosome is also highly dependent on the nature of the upstream IRES. Delivery of the ribosome to this AUG in an initiation-competent manner is particularly efficient and accurate with the EMCV IRES. PMID- 10376877 TI - The roles of Rrp5p in the synthesis of yeast 18S and 5.8S rRNA can be functionally and physically separated. AB - The yeast nucleolar protein Rrp5p is the only known trans-acting factor that is essential for the synthesis of both 18S rRNA and the major, short form of 5.8S (5.8Ss) rRNA, which were thought to be produced in two independent sets of pre rRNA processing reactions. To identify domains within Rrp5p required for either processing pathway, we have analyzed a set of eight deletion mutants that together cover the entire RRP5 sequence. Surprisingly, only one of the deletions is lethal, indicating that regions encompassing about 80% of the protein can be removed individually without disrupting its essential biological function. Biochemical analysis clearly demonstrated the presence of two distinct functional domains. Removal of each of three contiguous segments from the N-terminal half specifically inhibits the formation of 5.8Ss rRNA, whereas deleting part of the C terminal region of the protein only blocks the production of 18S rRNA. The latter phenotype is also caused by a temperature-sensitive mutation within the same C terminal region. The two functional regions identified by the mutational analysis appear to be correlated with the structural domains detected by computer analysis. They can even be physically separated, as demonstrated by the fact that full Rrp5p activity can be supplied by two contiguous protein fragments expressed in trans. PMID- 10376878 TI - Mutations in the MOF2/SUI1 gene affect both translation and nonsense-mediated mRNA decay. AB - Recent studies have demonstrated that cells have evolved elaborate mechanisms to rid themselves of aberrant proteins and transcripts. The nonsense-mediated mRNA decay pathway (NMD) is an example of a pathway that eliminates aberrant mRNAs. In yeast, a transcript is recognized as aberrant and is rapidly degraded if a specific sequence, called the DSE, is present 3' of a premature termination codon. Results presented here show that strains harboring the mof2-1, mof4-1, mof5-1, and mof8-1 alleles, previously demonstrated to increase the efficiency of programmed -1 ribosomal frameshifting, decrease the activity of the NMD pathway. The effect of the mof2-1 allele on NMD was characterized in more detail. Previous results demonstrated that the wild-type MOF2 gene is identical to the SUI1 gene. Studies on the mof2-1 allele of the SUI1 gene indicate that in addition to its role in recognition of the AUG codon during translation initiation and maintenance of the appropriate reading frame during translation elongation, the Mof2 protein plays a role in the NMD pathway. The Mof2p/Sui1 p is conserved throughout nature and the human homolog of the Mof2p/Sui1p functions in yeast cells to activate NMD. These results suggest that factors involved in NMD are general modulators that act in several aspects of translation and mRNA turnover. PMID- 10376879 TI - Sequence requirement for hand-in-hand interaction in formation of RNA dimers and hexamers to gear phi29 DNA translocation motor. AB - Translocation of DNA or RNA is a ubiquitous phenomenon. One intricate translocation process is viral DNA packaging. During maturation, the lengthy genome of dsDNA viruses is translocated with remarkable velocity into a limited space within the procapsid. We have revealed that phi29 DNA packaging is accomplished by a mechanism similar to driving a bolt with a hex nut, which consists of six DNA-packaging pRNAs. Four bases in each of the two pRNA loops are involved in RNA/RNA interactions to form a hexagonal complex that gears the DNA translocating machine. Without considering the tertiary interaction, in some cases only two G/C pairs between the interacting loops could provide certain pRNAs with activity. When all four bases were paired, at least one G/C pair was required for DNA packaging. The maximum number of base pairings between the two loops to allow pRNA to retain wild-type activity was five, whereas the minimum number was five for one loop and three for the other. The findings were supported by phylogenetic analysis of seven pRNAs from different phages. A 75-base RNA segment, bases 23-97, was able to form dimer, to interlock into the hexamer, to compete with full-length pRNA for procapsid binding, and therefore to inhibit phi29 assembly in vitro. Our result suggests that segment 23-97 is a self-folded, independent domain involved in procapsid binding and RNA/RNA interaction in dimer and hexamer formation, whereas bases 1-22 and 98-120 are involved in DNA translocation but dispensable for RNA/RNA interaction. Therefore, this 75-base RNA could be a model for structural studies in RNA dimerization. PMID- 10376880 TI - Transient interaction of BBP/ScSF1 and Mud2 with the splicing machinery affects the kinetics of spliceosome assembly. AB - Removal of introns from pre-mRNA is an essential step of gene expression. The splicing reaction is catalyzed in a large complex termed the spliceosome. Introns are recognized during the early steps of spliceosome assembly with the formation of commitment complexes. Intron recognition is mediated by the interaction of splicing factors with conserved sequences present in the pre-mRNA. BBP/SF1 participates in this recognition by interacting with the pre-mRNA branch point in both yeast and mammals. This protein, which is essential in yeast, also interacts with the U2AF65/Mud2 splicing factor. However, its precise role in splicing complex formation is still unclear. We have now analyzed the presence of BBP and Mud2 in yeast splicing complexes using supershift and coprecipitation assays. We found that BBP is present together with Mud2 in commitment complex 2 (CC2), but is not detectable in commitment complex 1 (CC1). Furthermore, genetic and biochemical depletion of BBP demonstrated that it is required for CC2 formation. In addition we observed that BBP and Mud2 are not detectable in pre-spliceosomes. These are the first commitment complex components that are shown to be released during or immediately after pre-spliceosome formation. Interestingly, depletion of BBP or disruption of MUD2 had no significant effect on pre-spliceosome formation and splicing in vitro but led to a transient accumulation of CC1. These observations support a model in which BBP and Mud2 are recycled during transition from CC2 to pre-spliceosome. PMID- 10376882 TI - The prevalence of Giardia and Cryptosporidium in Taiwan water supplies. AB - Giardia and Cryptosporidium have emerged as waterborne pathogens of concern. Thirty-one water samples were collected from nine potable water treatment plants in Taiwan and investigated for the presence of Giardia cysts and Cryptosporidium oocysts. The immunofluorescence assay was used for the simultaneous detection of cysts and oocysts. The frequency of occurrence of cysts was 77.8% for Giardia and 72.2% for Cryptosporidium in 18 raw water samples. Ten out of 13 samples collected from treated water samples showed the presence of cysts, while in 5 out of 13 treated water samples oocysts were detected. The risk assessment for adverse human effects arising from the presence of cysts and oocysts indicates the possibility of waterborne transmission of Giardia and Cryptosporidium infection in Taiwan if water is not adequately treated. PMID- 10376881 TI - Efficient reconstitution of functional Escherichia coli 30S ribosomal subunits from a complete set of recombinant small subunit ribosomal proteins. AB - Previous studies have shown that the 30S ribosomal subunit of Escherichia coli can be reconstituted in vitro from individually purified ribosomal proteins and 16S ribosomal RNA, which were isolated from natural 30S subunits. We have developed a 30S subunit reconstitution system that uses only recombinant ribosomal protein components. The genes encoding E. coli ribosomal proteins S2 S21 were cloned, and all twenty of the individual proteins were overexpressed and purified. Reconstitution, following standard procedures, using the complete set of recombinant proteins and purified 16S ribosomal RNA is highly inefficient. Efficient reconstitution of 30S subunits using these components requires sequential addition of proteins, following either the 30S subunit assembly map (Mizushima & Nomura, 1970, Nature 226:1214-1218; Held et al., 1974, J Biol Chem 249:3103-3111) or following the order of protein assembly predicted from in vitro assembly kinetics (Powers et al., 1993, J MoI Biol 232:362-374). In the first procedure, the proteins were divided into three groups, Group I (S4, S7, S8, S15, S17, and S20), Group II (S5, S6, S9, Sll, S12, S13, S16, S18, and S19), and Group III (S2, S3, S10, S14, and S21), which were sequentially added to 16S rRNA with a 20 min incubation at 42 degrees C following the addition of each group. In the second procedure, the proteins were divided into Group I (S4, S6, S11, S15, S16, S17, S18, and S20), Group II (S7, S8, S9, S13, and S19), Group II' (S5 and S12) and Group III (S2, S3, S10, S14, and S21). Similarly efficient reconstitution is observed whether the proteins are grouped according to the assembly map or according to the results of in vitro 30S subunit assembly kinetics. Although reconstitution of 30S subunits using the recombinant proteins is slightly less efficient than reconstitution using a mixture of total proteins isolated from 30S subunits, it is much more efficient than reconstitution using proteins that were individually isolated from ribosomes. Particles reconstituted from the recombinant proteins sediment at 30S in sucrose gradients, bind tRNA in a template-dependent manner, and associate with 50S subunits to form 70S ribosomes that are active in poly(U)-directed polyphenylalanine synthesis. Both the protein composition and the dimethyl sulfate modification pattern of 16S ribosomal RNA are similar for 30S subunits reconstituted with either recombinant proteins or proteins isolated as a mixture from ribosomal subunits as well as for natural 30S subunits. PMID- 10376883 TI - A study on oxidative stress in lead-exposed workers. AB - To explore the possible oxidative stress induced by lead, heparinized whole blood and urine of 66 secondary smelter lead workers (46 for Comet assay) and 28 controls were collected. The concentrations of blood lead (BPb) and urinary lead (UPb) and alpha-aminolevulinic acid (alpha-ALA), indices of lead exposure level of the body, were determined. Malondialdehyde (MDA) concentrations and superoxide dismutase (SOD) activity of plasma were also measured. Single-cell gel (SCG, Comet assay) was used to measure the DNA damage of peripheral blood cells. There was a positive correlation between the presence of Pb in blood and significant increases in MDA levels and SOD activity. Alcohol consumption and smoking with increased exposure to Pb was associated with enhanced DNA damage. A positive correlation was found between MDA and DNA damage. PMID- 10376885 TI - Effects of bis(4-chlorophenyl) sulfone on rats following 28-day dietary exposure. AB - The short-term oral toxicity of a recently identified environmental pollutant, bis(4-chlorophenyl) sulfone (BCPS), was studied. Groups of male Sprague-Dawley rats (n = 6) were administered BCPS via the diet at 0 (control), 10, 100, or 1000 ppm for 4 wk. Additional control and 1000 ppm groups were also treated for 1, 2, and 3 wk. At termination, high-dose animals showed depressed growth rate and food consumption, and 1 high dose animal in each of the wk-1, -3, and -4 groups had marked hematuria. Increased liver to body weight ratio was present at 100 ppm and increased kidney to body weight ratio at 1000 ppm. Marked increases in hepatic benzoylresorufin O-dealkylase (BROD) and pentoxyresorufin O-dealkylase (PROD) activities were detected starting at 10 ppm. There was a significant decrease in methoxyresorufin O-dealkylase (MROD) activity at 1000 ppm. Hepatic UDP glucuronosyltransferase (UDPGT) and glutathione S-transferase (GST) activities also increased starting at 100 ppm. A marked increase in urinary excretion of ascorbic acid was apparent starting at 10 ppm, while there were no changes in urinary N-acetylglucosaminidase (NAG) activity and protein levels. A threefold increase in serum cholesterol and a 30% increase in platelet counts were observed in the 1000 ppm group. Levels of thiobarbituric acid-reactive substances (TBARS) were increased by threefold in the liver of the high-dose animals but were not significantly altered in the serum. Tissue BCPS concentrations were dose dependent and followed the order: adipose tissue >>> liver > kidneys > brain, spleen, lungs. In the time-course study involving the control and high-dose groups, most of the treatment effects were clearly present in wk 1, and the severity of the effects remained at more or less the same levels thereafter. The exceptions were hepatic BROD and PROD activities, which showed a trend toward further increases with time of treatment. Liver and adipose tissue concentrations of BCPS remained unchanged from wk 1 to wk 4, while kidney concentrations increased with time. The results indicated that BCPS produced hepatic effects at the lowest dose level tested (10 ppm in the diet or 0.8 mg/kg/d). PMID- 10376884 TI - Iron exacerbates aniline-associated splenic toxicity. AB - Our earlier studies have shown that aniline exposure in rats causes time- and dose-dependent accumulation of iron in the spleen, which may exacerbate aniline splenotoxicity by catalyzing free-radical reactions. The present studies were conducted to test whether aniline-induced splenic toxicity could be potentiated by iron overload. For 30 d male Sprague-Dawley rats received the following treatments: 0.5 mmol/kg/d aniline hydrochloride (AH) by gavage (AH group); 3% carbonyl iron-supplemented diet (IR group); 0.5 mmol/kg/d AH by gavage and iron supplemented diet (AH + IR group); or no treatments (controls). Treatment-related significant increases in total iron, low molecular weight chelatable iron, lipid peroxidation, and protein oxidation were observed in the spleens of all the groups compared to control. However, these changes were much greater in the combined AH + IR group. The aniline-induced morphological changes in the spleen were consistent with our earlier observations, but were more pronounced in the AH + IR group. The increased toxicity, as evident from greater oxidative stress and morphological changes in the AH + IR group, suggests that iron potentiates the splenic toxicity of aniline. PMID- 10376886 TI - Distribution and metabolism of (5-hydroxymethyl)furfural in male F344 rats and B6C3F1 mice after oral administration. AB - (5-Hydroxymethyl)furfural (HMF), a heat-induced decomposition product of hexoses, is present in food and drink. Recent reports have shown HMF to be an in vitro mutagen after sulfate conjugation and to be a promoter as well as a weak initiator of colonic aberrant foci in rats. In order to investigate the metabolic activation further and to provide information for HMF toxicology studies, the disposition of [14C]-HMF has been investigated in male F344 rats and B6C3F1 mice following po administration of either 5, 10, 100, or 500 mg/kg. Tissue distribution results indicated that absorption of HMF was rapid in male rats and mice and that tissue concentrations in male mice at the earliest time point are not linearly proportional to dose. Excretion was primarily via the urine in both, with 60-80% of the administered dose excreted by this route in 48 h. Tissue/blood ratios of HMF-derived radioactivity were greater than 1 for liver and kidney. Three metabolites were identified and quantitated in urine. Formation of one of the metabolites, N-(5-hydroxymethyl-2-furoyl)glycine, was inversely proportional to dose in rats but not mice. None of the metabolites were sulfate conjugates nor likely to be formed from sulfate conjugates. There were relatively low levels of nonextractable radioactivity in liver, kidney, and intestines, indicating that some reactive intermediate(s) may be formed. PMID- 10376887 TI - Dependence of cadmium-metallothionein nephrotoxicity on glutathione. AB - Acute cadmium-metallothionein (CdMT) injection is frequently used as a model to study the mechanism of chronic Cd-induced nephrotoxicity. The purpose of this study was to investigate the relationship between glutathione (GSH) status and the ability of CdMT, either administered as a bolus dose or infused over a 24-h period by an osmotic minipump, to cause nephrotoxicity. GSH levels were modulated by pretreatment with either buthionine sulfoximine (BSO) or GSH. BSO enhanced while GSH suppressed acute CdMT nephrotoxicity. An infused dose of CdMT (150 microg Cd/kg) that was well tolerated when delivered over a 24-h period became nephrotoxic when GSH synthesis was inhibited by BSO. With depletion of GSH, as little as 0.4 microg Cd/g renal cortex was sufficient to cause nephrotoxicity after an acute dose of CdMT. While BSO had no effect on renal Cd accumulation, pretreatment with GSH reduced renal cortical Cd accumulation by 36%. CdMT nephrotoxicity was enhanced by depleting renal GSH, but without increasing renal Cd accumulation, which suggests that intracellular GSH is directly involved in protection against CdMT nephrotoxicity. Reduced Cd accumulation in the renal cortex following GSH pretreatment suggests an additional extracellular mechanism of GSH protection. It is concluded that GSH status is an important determinant of CdMT nephrotoxicity, with low GSH levels enhancing and high GSH levels reducing its toxicity, and that the mechanism appears to involve both intracellular and extracellular sites. PMID- 10376888 TI - Efficacy of topically administered doramectin against eyeworms, lungworms, and gastrointestinal nematodes of cattle. AB - OBJECTIVE: To evaluate efficacy of topically administered doramectin against eyeworms, lungworms, and gastrointestinal nematodes of cattle. ANIMALS: 400 cattle (20 cattle in each of 20 trials). PROCEDURE: Trials were conducted in North America; natural and experimentally induced infections were used. In each trial, cattle were allocated randomly to control (placebo [saline [0.9% NaCl] solution at 1 ml/10 kg of body weight] or untreated; n = 10) or doramectin treated (500 microg/kg of body weight; 10) groups. Treatments were applied in a single passage along the midline of the back, from the withers to the tailhead. Cattle were euthanatized > or =14 days after treatment, and worm burdens were determined by use of standard techniques. RESULTS: Efficacy of doramectin was > or =95.3% against adults of Thelazia gulosa, T skrjabini, Dictyocaulus viviparus, Haemonchus contortus, H placei, Ostertagia lyrata, O ostertagi, Trichostrongylus axei, Bunostomum phlebotomum, Capillaria spp, Cooperia oncophora, C pectinata, C punctata, C spatulata, C surnabada, Nematodirus spathiger, Strongyloides papillosus, T colubriformis, Oesophagostomum radiatum, and Trichuris spp. Efficacy was 95.1% against fourth-stage larvae of D viviparus, H placei, O lyrata, O ostertagi, T axei, C oncophora, C punctata, C spatulata, C surnabada, N helvetianus, T colubriformis, O radiatum, and Trichuris spp. In addition, efficacy against inhibited fourth-stage larvae of O ostertagi and Ostertagia spp was > or =98.1%. CONCLUSIONS AND CLINICAL RELEVANCE: A single topical application of doramectin pour-on was efficacious against a broad range of nematode species in cattle. PMID- 10376889 TI - Superoxide production in phagocytes obtained from Mycobacterium marinum stimulated goldfish (Carassius auratus) that were exposed to copper. AB - OBJECTIVE: To investigate the effects of copper exposure and recovery from copper toxicosis on the nonspecific immune response in Mycobacterium marinum-inoculated goldfish. ANIMALS: Goldfish (Carassius auratus) with a mean weight of 33.5 g. PROCEDURE: Superoxide (O2-) production was measured in fish 2 to 6 weeks after injection with phosphate-buffered saline (PBS) solution or M marinum (10(2) to 10(7) colony-forming units [CFU]/fish). Then, paired groups of fish were injected with PBS solution or 10(4) CFU of M marinum and exposed to copper (100 microg/L) for 7 days or for 4 days with 3 days of recovery. One paired group not exposed 14 days later to copper served as control fish. Phagocyte production of O2-was measured by use of the nitroblue tetrazolium reduction assay. Inflammation and bacterial colony counts were determined by use of routine histologic and microbiologic procedures. RESULTS: Superoxide production achieved a maximal response 2 to 4 weeks after M marinum inoculation. Compared with control fish, O2 production increased in the groups exposed to copper but then decreased in the exposed groups that were allowed to recover. Superoxide response and peritoneal inflammation were greater in M marinum-inoculated groups than in non-inoculated groups. CONCLUSIONS: Copper exposure and inoculation with M marinum increased O2- production, whereas recovery after exposure decreased O2- production, even in fish that were immunostimulated by M marinum. CLINICAL RELEVANCE: When the antimicrobial oxidative response is suppressed after copper exposure, steps should be taken to avoid imposing additional stress and minimize the possibility of resurgent or secondary pathogenic infections. PMID- 10376890 TI - Stress leak point pressures and urethral pressure profile tests in clinically normal female dogs. AB - OBJECTIVE: To develop a stress leak point pressure (LPP) test for dogs, determine LPP for continent female dogs, and determine urethral pressure profile (UPP) values for nonanesthetized, continent female dogs. ANIMALS: 22 continent female dogs weighing from 21 to 29 kg. PROCEDURE: A standard UPP test and a modification of the LPP test used in women were performed on all dogs. On 3 occasions, dogs underwent UPP testing while awake. They then were anesthetized with propofol, and LPP was measured at bladder volumes of 75, 100, and 150 ml. For LPP tests, abdominal pressure was applied by inflating a human blood pressure cuff placed around the dog's abdomen. LPP were recorded through a urethral catheter (bladder LPP) and a rectal balloon catheter (abdominal LPP). RESULTS: Mean +/- SD and median maximal urethral closure pressure was 110.1+/-20.2 and 109.0 cm water, respectively. Mean bladder LPP for the 75, 100, and 150 ml bladder volumes was 172.4 cm water. Significant differences among LPP for the 3 bladder volumes were not detected. CONCLUSIONS: Stress LPP can be recorded in female dogs. PMID- 10376891 TI - Interday variation in vertical ground reaction force in clinically normal Greyhounds at the trot. AB - OBJECTIVE: To determine the effect of interday variation on vertical ground reaction force variables in dogs. ANIMALS: 52 clinically normal Greyhounds of either sex weighing between 22 and 35 kg. PROCEDURE: Dogs were led at a trot across a floor-mounted force platform to determine vertical ground reaction force variables (peak [PFz] and impulse [IFz]) from hind limbs. Data were collected from each dog on 3 consecutive days. Variance components were estimated, using maximal likelihood to evaluate contributions of interday variation within dogs and variation attributable to dogs and repetitions. An ANOVA was used to test significance of interday variation within dogs and day within dog interactions. RESULTS: PFz, IFz, or both differed significantly from day to day for 29 of 52 dogs. Only PFz differed significantly among days for 16 dogs, and only IFz differed among days for 5 dogs. The PFz and IFz differed significantly from day to day in 8 dogs. Using ANOVA, the difference for PFz and IFz among days within dogs was significant. CONCLUSIONS: Effect of day within dog variation (interaction) should be considered as a component in statistical models in which data from 1 day are evaluated against data from the same subject on another day. We propose a statistical model that incorporates an accommodation for interday variation. Investigators should determine the factors that affect their studies, including the extent of interday variation, and compensate for the variation attributable to each factor in the statistical models used to analyze their data. PMID- 10376892 TI - In vitro contractile responses and contracture testing of skeletal muscle from Quarter Horses with exertional rhabdomyolysis. AB - OBJECTIVE: To determine whether increased sensitivity to pharmacologic agents was a general property of equine exertional myopathies, including polysaccharide storage myopathy (PSSM) in Quarter Horses. ANIMALS: 5 adult Quarter Horses with exertional rhabdomyolysis and abnormal polysaccharide accumulation in skeletal muscle and 4 clinically normal adult Quarter or Quarter-type horses. PROCEDURES: Twitch time course measurements and contracture responses to various concentrations of caffeine and halothane for small bundles of intact external intercostal muscle fibers were measured in all horses. RESULTS: Caffeine contracture threshold of muscles from Quarter Horses with PSSM was not different from that of clinically normal horses (5 mM in both groups). Muscles from horses with PSSM and from clinically normal horses did not have contracture in response to up to 2% halothane. CONCLUSIONS AND CLINICAL RELEVANCE: Results were in contrast to the increased sensitivity to caffeine and halothane for muscles from Thoroughbreds with recurrent exertional rhabdomyolysis (RER). Although clinical signs of muscular stiffness after exercise are similar between Quarter Horses with PSSM and Thoroughbreds with RER, these breeds appear to have 2 distinct myopathies with different pathophysiologic bases. Unlike RER in Thoroughbreds, PSSM in Quarter Horses does not appear to be accompanied by a defect in regulation of muscle contraction. PMID- 10376893 TI - Aerosolized albuterol sulfate used as a bronchodilator in horses with recurrent airway obstruction. AB - OBJECTIVE: To determine the dose of aerosolized albuterol sulfate required to cause bronchodilation in horses with recurrent airway obstruction (RAO) and duration of this effect. ANIMALS: 19 horses with RAO (10 in experiment 1; 9 in experiment 2). PROCEDURE: Horses were moved from pasture to stables, and airway obstruction was induced. Pulmonary function was measured in 10 horses before and 5, 10, and 30 minutes after administration of vehicle or 120, 240, 360, or 720 microg of the drug. Nine horses received vehicle or 360 or 720 microg of albuterol, and pulmonary function was measured at baseline and 5 minutes and 1, 2, 3, 4, 5, 6, and 7 hours later. Horses were evaluated for adverse drug effects. RESULTS: 360 microg of albuterol was required to cause significant bronchodilatation; 720 microg did not enhance bronchodilatation or increase duration of action. Depending on which pulmonary function parameter was evaluated, bronchodilatation achieved by use of albuterol lasted between 30 minutes and 1 hour. Because there was a significant vehicle effect, the combined effect of vehicle and drug lasted up to 3 hours. Adverse effects were not observed. CONCLUSIONS: Aerosolized albuterol, 360 or 720 microg, is a safe and effective bronchodilator in horses with RAO. Onset of action is rapid (5 minutes), and effects last from 30 minutes to 3 hours. CLINICAL RELEVANCE: Aerosolized albuterol is useful for treatment of bronchospasm in horses with RAO. PMID- 10376894 TI - Serologic cross-reactivity of antibodies against Borrelia theileri, Borrelia burgdorferi, and Borrelia coriaceae in cattle. AB - OBJECTIVE: To evaluate the immune response induced by Borrelia theileri infection and to determine whether B theileri induces cross-reacting antibodies to other bovine borreliae. ANIMALS: Two 3-month-old calves, 1 of which was splenectomized. PROCEDURE: Calves were exposed to Boophilus microplus infected with B theileri. Rectal temperature, PCV, bacteremia, and clinical signs of infection were monitored. Serum was obtained weekly and used to evaluate the humoral response to homologous antigen and B burgdorferi and B coriaceae, using an indirect fluorescent antibody (IFA) test, and to B burgdorferi, using a commercially available ELISA. The identity of cross-reacting antigens was explored, using monoclonal antibodies to genus- and species-specific antigens in an IFA test. RESULTS: B theileri-infected calves produced antibodies that cross-reacted with B burgdorferi and B coriaceae whole-cell antigens. Borrelia theileri whole-cell antigen was recognized by genus-specific monoclonal antibody H9724 but not by species-specific antibody H5332. False-positive reactions were not observed when serum from B theileri-infected calves was tested by use of the ELISA for B burgdorferi. CONCLUSIONS: B theileri induces humoral responses in infected cattle that can be confused with those of other borrelial infections. Care must be taken to definitively distinguish between the various borreliae that may cause disease in cattle. CLINICAL RELEVANCE: Serologic cross-reactivity must be taken into account when making a serodiagnosis of Lyme borreliosis or epizootic abortion in epidemiologic studies involving cattle. PMID- 10376895 TI - Effect of alternate-day prednisolone administration on hypophyseal-adrenocortical activity in dogs. AB - OBJECTIVE: To evaluate effect of alternate-day oral administration of prednisolone on endogenous plasma ACTH concentration and adrenocortical response to exogenous ACTH in dogs. ANIMALS: 12 Beagles. PROCEDURE: Dogs were allotted to 2 groups (group 1, 8 dogs treated with 1 mg of prednisolone/kg of body weight; group 2, 4 dogs given excipient only). During a 30-day period, blood samples were collected for determination of plasma ACTH and cortisol concentrations before, during, and after treatment with prednisolone. From day 7 to 23, prednisolone or excipient was given on alternate days. Sample collection (48-hour period with 6 hour intervals) was performed on days 1, 7, 15, 21, and 28; on other days, sample collection was performed at 24-hour intervals. Pre- and post-ACTH plasma cortisol concentrations were determined on days 3, 9, 17, 23, and 30. RESULTS: A significant difference was detected between treatment and time for group 1. Plasma ACTH concentrations significantly decreased for 18 to 24 hours after prednisolone treatment in group-1 dogs. At 24 to 48 hours, ACTH concentrations were numerically higher but not significantly different in group-1 dogs. Post ACTH plasma cortisol concentration significantly decreased after 1 dose of prednisolone and became more profound during the treatment period. However, post ACTH cortisol concentration returned to the reference range 1 week after prednisolone administration was discontinued. CONCLUSIONS AND CLINICAL RELEVANCE: Single oral administration of 1 mg of prednisolone/kg significantly suppressed plasma ACTH concentration in dogs for 18 to 24 hours after treatment. Alternate day treatment did not prevent suppression, as documented by the response to ACTH. PMID- 10376896 TI - Effect of intramammary infusion of beta-1,3-glucan or interleukin-2 on leukocyte subpopulations in mammary glands of sheep. AB - OBJECTIVE: To investigate effects of intramammary infusion of beta-1,3-glucan or recombinant ovine interleukin-2 (rOvIL-2) on blood and mammary leukocyte subpopulations and their expression of various surface antigens in sheep. ANIMALS: 12 healthy multiparous, nonpregnant, nonlactating fine-wool Merino ewes. PROCEDURE: Beta-1,3-glucan in pyrogen-free saline solution (PFSS; n = 6), rOvIL-2 in PFSS (3), or PFSS (3) was infused on days 0 and 7 in 1 udder half of each ewe. Jugular vein blood and mammary secretion samples were taken before infusion and on days 2, 7, and 21 after infusion. Total and differential leukocyte counts were obtained, and blood and mammary cells were labeled for flow cytometry. RESULTS: Slight swelling of the mammary glands was observed on day 2 after rOvIL-2 but not after beta-1,3-glucan infusion. Both substances induced significant increase in mammary secretion leukocyte numbers, compared with controls. Beta-1,3-Glucan induced an influx of monocyte/macrophages, whereas neutrophils were the predominating cell population after rOvIL-2 infusion. Beta-1,3-glucan induced selection for CD4+, B-cell+, WC1+, and L-selectin+ lymphocytes on day 14 after infusion. By comparison, rOvIL-2 induced selection for B-cell+ and L-selectin+ lymphocytes on days 14 and 21 and depletion of CD8 and, to some degree, of IL-2R+ lymphocytes. Beta-1,3-Glucan induced an increase in the proportion of CD14+ leukocytes, indicating selective migration of monocyte/ macrophages to the nonlactating udder. CONCLUSION: Beta-1,3-Glucan and rOvIL-2 can modulate nonspecific immunity in the udder of sheep but may exert their effects by differing mechanisms. Clinical Relevance-Stimulation of the nonspecific defense against udder infections may improve control of mastitis. PMID- 10376897 TI - Evaluation of 2-deoxy-D-glucose for induction of a stress response in pigs. AB - OBJECTIVE: To determine the most effective route and dose for 2-deoxy-D-glucose (2DG) administration in swine, kinetics of 2DG, endogenous glucose concentration in the blood, effects of 2DG on cortisol concentration, and effects of 2DG administration in vivo on lymphocyte proliferation in vitro. ANIMALS: 14 Salmonella-free male and female mixed-breed pigs. PROCEDURE: A cannula was inserted in the femoral artery of each pig to allow for frequent blood collection with minimal external stress. The concentration and duration of 2DG in the blood was monitored while varying dose (250, 500, or 750 mg/kg of body weight) and route (IV, SC, IM, or IP) of 2DG administration. Blood samples were collected at various time points and assayed for lymphocyte response to concanavalin A and cortisol, endogenous glucose, and 2DG concentrations. RESULTS: The 2 best routes for administration of 2DG were IV and SC. If the IV route was chosen, the optimal dose was 500 mg of 2DG/kg; the optimal dose for SC administration was 750 mg/kg. CONCLUSIONS: 2DG induces a stress response in pigs similar to that in rodents. The use of 2DG in a porcine stress model should be effective for studying the possible role of stress in the pathogenesis and shedding of microorganisms. PMID- 10376898 TI - Nitric oxide synthase activity in healthy and interleukin 1beta-exposed equine synovial membrane. AB - OBJECTIVE: To quantitate nitric oxide synthase (NOS) activity in healthy and interleukin 1beta (IL-1beta)-exposed equine synovial membrane. ANIMALS: 6 healthy horses, 2 to 8 years old. PROCEDURE: Recombinant human IL-1beta (0.35 ng/kg of body weight) was injected intra-articularly into 1 metacarpophalangeal joint of each horse. The contralateral joint served as an unexposed control. All horses were euthanatized 6 hours after injection of IL-1beta, and synovial membrane specimens were assayed for NOS activity by measuring conversion of arginine to citrulline. Severity of inflammation was semiquantitated by analysis of synovial fluids and histologic examination of synovial membrane. RESULTS: Equine synovial membrane had minimal NOS activity. A significant difference was not detected in NOS activity between control and IL-1beta-exposed specimens. Histologic examination revealed a neutrophilic infiltrate in synovial membrane exposed to IL 1beta. Synovial fluid from IL-1beta-exposed joints had a moderate inflammatory response and significantly greater concentrations of IL-1beta and interleukin-6 than fluid from healthy joints. CONCLUSION: Healthy equine synovial membrane had low NOS activity that was not affected by exposure to IL-1beta. PMID- 10376899 TI - Influence of sotalol on the time constant of isovolumic left ventricular relaxation in anesthetized dogs. AB - OBJECTIVE: To determine the effects of various doses of sotalol on myocardial relaxation in healthy dogs. ANIMALS: 12 healthy adult mixed-breed dogs anesthetized with thiopental and alpha-chloralose. PROCEDURES: Left ventricular pressure (LVP), aortic pressure, and aortic flow velocity were measured. The time constant of isovolumic relaxation (tau) was determined by means of linear regression of the natural logarithm of LVP and by means of direct measurement from the LVP-versus-time curve. Sotalol was administered IV at cumulative doses of 1, 2, 4, and 8 mg/kg to 6 dogs; the other 6 were used as controls. Mean systolic aortic pressure was used to assess afterload, and maximal rate of increase in LVP versus time (dP/dt) was used as an index of contractility. RESULTS: After administration of the first dose of sotalol, tau was increased significantly, and maximum dP/dt was decreased significantly, compared with baseline values. Administration of additional doses of sotalol did not result in any additional change in tau, but maximum dP/dt increased, and maximum dP/dt after administration of the final dose of sotalol was significantly higher than maximum dP/dt after administration of the first dose. There were no significant changes in mean systolic aortic pressure. CONCLUSIONS AND CLINICAL RELEVANCE: In healthy dogs, sotalol did not have any negative effect on myocardial relaxation beyond those attributable to its beta-blocking properties, despite an increase in intracellular ionized calcium concentration, as suggested by an increase in maximum dP/dt after an initial decrease. PMID- 10376900 TI - Biosafety of Brucella abortus strain RB51 for vaccination of mature bulls and pregnant heifers. AB - OBJECTIVE: To determine shedding and colonization profiles in mature sexually intact bulls and pregnant heifers after vaccination with a standard calfhood dose of Brucella abortus strain RB51 (SRB51). ANIMALS: 6 sexually mature 3-year-old Jersey bulls and 7 mixed-breed heifers in midgestation. PROCEDURE: Bulls and pregnant heifers were vaccinated IM with the standard calfhood dose of 3x10(10) colony-forming units of SRB51. After vaccination, selected body fluids were monitored weekly for vaccine organism shedding. Pathogenesis was monitored in bulls by weekly breeding soundness examination and, in heifers, by delivery status of the calf. Vaccine organism colonization was assessed by obtaining select tissues at necropsy for bacterial culture. Serologic analysis was performed by use of numerous tests, including complement fixation, an SRB51-based ELISA, and immunoblot analysis. RESULTS: After vaccination, none of the vaccinated bulls or heifers shed SRB51 in their secretions. Results of breeding soundness examination for bulls were normal as was delivery status of the pregnant heifers (6 live births, 1 dystocia). At necropsy, SRB51 was not recovered from any of the selected tissues obtained from bulls, heifers, or calves; however, serologic analysis did detect SRB51-specific antibodies in all cattle. CONCLUSIONS AND CLINICAL RELEVANCE: Vaccination with the standard calfhood dose of SRB51 administered IM was not associated with shedding or colonization in sexually mature bulls or pregnant heifers. Also, under conditions of this study with small numbers of animals, IM vaccination with SRB51 does not appear to cause any reproductive problems when administered to sexually mature cattle. PMID- 10376901 TI - Genomic screening for fucosidosis in English Springer Spaniels. AB - OBJECTIVE: To develop a robust molecular genetic test for alpha-L-fucosidosis in English Springer Spaniels and to screen dogs from the United Kingdom and United States for the mutant allele. ANIMALS: 35 English-bred English Springer Spaniels, 60 American-bred English Springer Spaniels, and 1 affected dog and its parents from a family of English Springer Spaniels in Colorado. PROCEDURE: Polymerase chain reaction analysis was used to amplify the mutated region in the gene encoding alpha-L-fucosidase. High guanine-cytosine (GC) content of the region required use of an amplification buffer with high pH. Mutant and normal alleles were separated by polyacrylamide gel electrophoresis. Molecular genetic test results were compared with enzyme data. RESULTS: A 262-bp PCR product was amplified from normal dogs and compared with a 248-bp product from affected dogs. Carriers had 1 copy of each allele, distinguishable by the 14-bp size difference. Two carriers among the English-bred dogs were identified by use of enzyme and genomic DNA analyses. The molecular defect in dogs from Colorado was proven to be the same as that in British and Australian dogs. None of the other 60 American bred dogs carried the mutant allele. CONCLUSIONS AND CLINICAL RELEVANCE: A PCR method that can be used to identify dogs affected with or carriers of the autosomal recessive disease fucosidosis was established. Amplification was achieved within a GC-rich region, using a method that may be useful in overcoming amplification problems in GC-rich areas within other genes. Using this test, fucosidosis can be controlled and ultimately eradicated from the English Springer Spaniel population. PMID- 10376902 TI - Effect of hypothyroidism on blood lipid concentrations in horses. AB - OBJECTIVE: To measure and compare concentrations of selected blood lipids before and after thyroidectomy in horses. ANIMALS: 5 healthy adult mares. PROCEDURE: Mares were confirmed to be euthyroid. Thyroidectomy was performed, and hypothyroidism was confirmed. Selected blood lipid variables were measured before hypothyroidism was induced and weekly for 4 weeks after induction. Plasma concentrations of very low-density lipoprotein (VLDL), low-density lipoprotein (LDL), serum triglyceride (TG), total cholesterol (TC), and nonesterified fatty acid (NEFA) were measured. The composition of VLDL and LDL also was examined. RESULTS: Mean plasma concentrations of VLDL and LDL increased significantly after thyroidectomy. By 4 weeks after thyroidectomy, a ninefold increase in mean plasma concentration of VLDL and a threefold increase in LDL, compared with baseline values, were detected. After thyroidectomy, mean percentage of TG in VLDL increased significantly, whereas free cholesterol and cholesterol ester content decreased. Mean percentage of TG in LDL also increased by 3 to 4 weeks after thyroidectomy. Serum concentrations of TG and TC increased, whereas serum NEFA concentration decreased. CONCLUSIONS: Hypothyroidism significantly alters blood lipid concentrations of horses. After thyroidectomy, markedly high VLDL concentration, appearance of TG-rich VLDL, increased serum concentrations of TG and TC, and decreased blood concentration of NEFA were evident. CLINICAL RELEVANCE: Examination of blood lipid concentrations of horses may be useful for detecting naturally acquired hypothyroidism. PMID- 10376903 TI - Development of a snapback method of single-strand conformation polymorphism analysis for genotyping Golden Retrievers for the X-linked muscular dystrophy allele. AB - OBJECTIVE: To develop a snapback method of single-strand conformation polymorphism (SSCP) analysis for genotyping Golden Retrievers for the X-linked muscular dystrophy allele. ANIMALS: 20 Golden Retriever puppies from a colony with X-linked muscular dystrophy. PROCEDURE: DNA spanning the canine dystrophin mutation was amplified by means of a polymerase chain reaction (PCR), using a primer modified to have an additional sequence at the 5' terminus. The primer was designed so that 1 terminus of the single-stranded PCR product could anneal to the normal sequence flanking the region of the mutation in the allele but not in the mutant allele. True disease status of the dogs was determined by means of a PCR and restriction digest protocol. RESULTS: Snapback SSCP analysis allowed for accurate and unambiguous genotyping of unaffected, carrier, and affected dogs, whereas conventional SSCP analysis, using the unmodified primer, did not. Creatine kinase activities measured within 24 hours after birth were not consistent with genotype. CONCLUSION AND CLINICAL RELEVANCE: Snapback SSCP analysis provided a simple, fast, and accurate method for genotyping Golden Retrievers for the mutation known to cause X-linked muscular dystrophy. PMID- 10376905 TI - Anesthesia of horses with a combination of detomidine, zolazepam, tiletamine, and isoflurane immediately after strenuous treadmill exercise. AB - OBJECTIVES: To evaluate effects of strenuous exercise in adult horses immediately before anesthesia and to determine whether prior exercise affects anesthesia induction, recovery, or both. ANIMALS: 6 healthy Thoroughbreds in good condition and trained to run on a treadmill, each horse serving as its own control. PROCEDURE: Horses ran on a treadmill until fatigued, then were sedated immediately with detomidine hydrochloride and anesthetized with a zolazepam hydrochloride-tiletamine combination. Anesthesia was maintained with isoflurane in oxygen for another 90 minutes. Blood samples were taken before, during, and after exercise and during anesthesia. RESULTS: During exercise, changes in heart rate, core body temperature, plasma lactate concentration, arterial pH, and PaCO2 were significant. Plasma ionized calcium concentration was lower after exercise, compared with baseline values, and remained lower at 30 minutes of isoflurane anesthesia. Compared with baseline values, plasma chloride concentration decreased significantly during anesthesia after exercise. Cardiac output during anesthesia was significantly lower than that during preexercise, but significant differences between experimental and control periods were not observed. Arterial blood pressure during anesthesia was significantly lower than that during preexercise and initially was maintained better during isoflurane anesthesia after exercise. Cardiac output and blood pressure values were clinically acceptable throughout anesthesia. CONCLUSION: Administration of detomidine hydrochloride followed by zolazepam hydrochloride-tiletamine appeared to be safe and effective for sedation and anesthesia of horses that had just completed strenuous exercise. CLINICAL RELEVANCE: Anesthetic given in accordance with this protocol can be used to anesthetize horses that are injured during athletic competition to assess injuries, facilitate first aid, and possibly allow salvage of injured horses. PMID- 10376904 TI - Pharmacokinetics of the antihyperglycemic agent metformin in cats. AB - OBJECTIVE: To determine the pharmacokinetics of metformin in healthy cats after single-dose IV and oral administration of the drug. ANIMALS: 6 healthy adult ovariohysterectomized cats. PROCEDURE: In a randomized cross-over design study, each cat was given 25 mg of metformin/kg of body weight, IV and orally. Blood and urine samples were collected after drug administration, and concentrations of metformin in plasma and urine were determined by use of high-performance liquid chromatography. RESULTS: Disposition of the drug was characterized by a three compartment model with a terminal phase half-life of (mean +/- SD) 11.5+/-4.2 hours. Metformin was distributed to a small central compartment of 0.057+/-0.017 L/kg and to 2 peripheral compartments with volumes of distribution of 0.12+/-0.02 and 0.37+/-0.38 L/kg. Steady-state volume of distribution was 0.55+/-0.38 L/kg. After IV administration, 84+/-14% of the dose was excreted unchanged in urine, with renal clearance of 0.13+/-0.03 L/h/kg; nonrenal clearance was negligible (0.02+/-0.02 L/kg). Mean bioavailability of orally administered metformin was 48%. CONCLUSIONS: The general disposition pattern of metformin in cats is similar to that reported for humans. Metformin was eliminated principally by renal clearance; therefore, this drug should not be used in cats with substantial renal dysfunction. CLINICAL RELEVANCE: On the basis of our results, computer simulations indicate that 2 mg of metformin/kg administered orally every 12 hours to cats will yield plasma concentrations documented to be effective in humans. PMID- 10376906 TI - Effects of oral administration of orotic acid on hepatic morphologic characteristics and serum biochemical variables in cats. AB - OBJECTIVE: To evaluate orotic acid (OA) as a possible etiologic factor in cats with idiopathic hepatic lipidosis (HL). ANIMALS: 20 clinically normal adult female cats. PROCEDURE: Cats were fed a control diet or a diet containing less protein. On day 1 of the control period, blood, urine, and liver biopsy specimens were obtained. Each cat was given an oral dose of water daily. On days 8, 15, and 22, blood and urine specimens were collected as on day 1. On day 29, liver, blood, and urine samples were obtained as on day 1. After a resting period of 30 to 60 days, cats were treated with orotic acid. Serum biochemical analyses, urinary OA-to-creatinine ratios, and liver biopsy specimens were evaluated. Cats were given OA orally (suspension or capsules) for 29 days. Sample collection and data obtained were identical to those described for the control period. RESULTS: Urinary OA-to-creatinine ratios were significantly higher in all treated cats, but ratios were significantly higher in those receiving OA in capsules than in those receiving OA in suspension. Diet or treatment did not alter hepatic biochemical or histologic variables significantly. However, 7 cats given the highest dose of OA in capsules developed azotemia, urolithiasis, and renal changes. CONCLUSIONS: Most concentrations of OA used in this study did not induce HL in cats during a 29-day period, but the highest dosage used did result in renal disease. CLINICAL RELEVANCE: Orotic acid does not appear to be involved in the genesis of HL in cats. PMID- 10376907 TI - Urinary orotic acid-to-creatinine ratios in cats with hepatic lipidosis. AB - OBJECTIVE: To determine urinary orotic acid (OA) concentration and evaluate the urinary OA-to-creatinine ratio (OACR) in cats with hepatic lipidosis (HL). ANIMALS: 20 cats with HL and 20 clinically normal cats. PROCEDURE: Hepatic lipidosis was diagnosed on the basis of clinical signs, results of serum biochemical analyses, exclusion of other concurrent illness, and cytologic or histologic evaluation of liver biopsy specimens. Urine samples were collected from each cat and frozen at -20 C until assayed. Urine creatinine concentrations were determined, using an alkaline picrate method followed by spectrophotometric assay. Urine OA concentration was determined, using high-performance liquid chromatography. Minimum amount of detectable OA in feline urine was 1 microg/ml. Because of small interfering peaks near the base of the OA peak, the minimum quantifiable concentration of OA was determined to be 5 microg/ml. Urinary OACR was compared in both groups of cats. RESULTS: Differences in urinary OACR were not detected between clinically normal cats and cats with HL. Peaks were not detected for urinary OA in any of the 20 clinically normal cats. Of the 20 HL cats, 14 did not have detectable peaks for urinary OA. Of the 6 HL cats that had detectable urinary OA peaks, 3 had values of <5 microg/ml. CONCLUSIONS: Apparently, OACR does not increase significantly in cats with HL. CLINICAL RELEVANCE: Urinary OACR is not a useful diagnostic test for HL in cats. PMID- 10376908 TI - Effects of glutamine supplementation of an amino acid-based purified diet on intestinal mucosal integrity in cats with methotrexate-induced enteritis. AB - OBJECTIVE: To determine effects of glutamine-supplemented and glutamine-free amino acid-based purified diets, compared with a dry expanded diet, on intestinal structure and function in a model that used cats with methotrexate-induced enteritis. ANIMALS: 18 adult specific-pathogen-free cats. PROCEDURE: 12 cats were given intragastric feedings of an amino acid-based purified diet supplemented with glutamine (7% [wt:wt]) or an isonitrogenous amount of glycine and alanine; 6 cats consumed a dry expanded diet. After 21 days, cats received methotrexate (MTX; 11 mg/kg of body weight, IV). Intestinal permeability testing was performed immediately before and 66 hours after MTX administration. Celiotomy was performed 72 hours after MTX administration for aseptic removal of mesenteric lymph nodes, collection of full-thickness intestinal biopsy specimens, determination of intestinal cellular proliferation, and collection of aortic and portal venous blood samples for determination of arteriovenous amino acid concentrations across the intestine. RESULTS: Administration of MTX was associated with severe enterotoxicosis manifested as diarrhea (8/12 cats), vomiting (12/12), and positive results for bacterial culture of mesenteric lymph nodes (12/12) in cats receiving the purified diets, independent of glutamine supplementation. Diet did not affect villus tip length and villus surface area in the small intestine or cellular proliferation. Administration of MTX was associated with significantly increased intestinal permeability, which was not attenuated by glutamine supplementation. CONCLUSIONS: Feeding of a glutamine-supplemented amino acid based purified diet was unable to preserve intestinal function in cats with MTX induced enteritis. CLINICAL RELEVANCE: Intestinal morphologic alterations correlate poorly with intestinal function as measured by means of bacterial translocation and intestinal permeability. PMID- 10376910 TI - Cardiorespiratory effects of a tiletamine/zolazepam-ketamine-detomidine combination in horses. AB - OBJECTIVE: To determine cardiorespiratory effects of a tiletamine/zolazepam ketamine-detomidine (TZKD) combination in horses. ANIMALS: 8 healthy adult horses. PROCEDURE: Horses were instrumented for measurement of cardiorespiratory, acid-base, and electrolyte values. Each horse was given xylazine (0.44 mg/kg of body weight, IV) 10 to 15 minutes prior to induction of recumbency by administration of the TZKD combination. Cardiorespiratory, acid-base, and electrolyte values were measured at 5-minute intervals for > or =30 minutes. RESULTS: All horses became recumbent within 1 minute after IV administration of TZKD. Mean +/- SD duration of recumbency was 40+/-8 minutes. All horses regained standing position after < or =2 attempts. Quality of anesthesia and analgesia was determined to be satisfactory in all horses. Xylazine induced decreases in respiratory rate, heart rate, cardiac output, maximum rate of increase of right ventricular pressure, and rate pressure product. The PaCO2, right atrial pressure, and peripheral vascular resistance increased, whereas blood temperature, PO2, pHa, HCO3-, PCV, total solids, Na, and K values remained unchanged. Subsequent administration of TZKD caused right atrial pressure and PaCO2 to increase and PaO2 to decrease, compared with values obtained after xylazine administration. Remaining cardiorespiratory, acid-base, hematologic, and electrolyte values did not differ from those obtained after xylazine administration. CONCLUSION: IV administration of TZKD induces short-term anesthesia in horses. Potential advantages of this drug combination are the small volume of drug administered; minimal cardiorespiratory depression; quality of induction and maintenance of, and recovery from, anesthesia; and duration of drug effects. PMID- 10376909 TI - Pulmonary distribution of aerosolized technetium Tc 99m pentetate after administration of a single dose of aerosolized albuterol sulfate in horses with recurrent airway obstruction. AB - OBJECTIVE: To determine whether pulmonary distribution of aerosolized technetium Tc 99m pentetate is improved after inhalation of a single dose of albuterol sulfate in horses susceptible to recurrent airway obstruction (heaves). ANIMALS: 6 horses with heaves and 4 horses with normal respiratory tract function. PROCEDURE: Images were obtained during ventilation of horses at baseline (maximal change in pleural pressure during tidal breathing [deltaPpImax] >15 cm H2O) and after aerosolized albuterol sulfate (360 microg) administration, with a 24-hour washout period between experiments. The deltaPpImax was determined prior to the baseline scan, prior to albuterol sulfate administration, and 5 minutes after albuterol sulfate administration. Images were assessed by visual inspection (semi quantitative scoring system) and histogram analysis. RESULTS: Images obtained from horses with heaves had nonuniform pulmonary distribution of radionuclide characterized by poor penetration in peripheral lung fields and excess deposition in large airways. Histogram analysis of images of the caudal portions of the lungs revealed nonuniform radionuclide deposition in horses with heaves and uniform radionuclide deposition in control horses. CONCLUSION: Administration of a single dose of aerosolized albuterol sulfate improved pulmonary distribution of aerosolized radiolabeled pentetate suspension in horses with heaves but did not alter pulmonary distribution in clinically normal horses. CLINICAL RELEVANCE: Precedent bronchodilator administration may improve pulmonary distribution of aerosolized, surface-active anti-inflammatory preparations. PMID- 10376911 TI - Scintigraphic, sonographic, and histologic evaluation of renal autotransplantation in cats. AB - OBJECTIVE: To determine scintigraphic, sonographic, and histologic changes associated with renal autotransplantation in cats. ANIMALS: 7 adult specific pathogen-free cats: 5 males, 2 females, 1 to 9 years old. PROCEDURE: Renal autotransplantation was performed by moving a kidney (5 left, 2 right) to the left iliac fossa. Before and at multiple times after surgery, for a total of 28 days, cats were evaluated by B-mode and Doppler ultrasonography, scintigraphy, and renal biopsy. RESULTS: By 24 hours after surgery, a significant decrease (42%) in mean glomerular filtration rate (GFR) and an increase in mean renal size (81% increase in cross-sectional area) were evident in the transplanted kidney, compared with preoperative values. By postsurgery day 28, reduction in GFR was 23%. Significant changes in renal blood flow velocity were identified in both kidneys. Consistent changes in resistive index or pulsatility index for either kidney could not be identified. When all postoperative histologic data were combined, the histologic score, indicating degree and numbers of abnormalities detected, for the transplanted kidney was significantly higher than that for the control kidney. CONCLUSIONS: Significant changes in renal function, size, and histologic abnormalities develop secondary to acute tubular necrosis in cats after uncomplicated renal autotransplantation. CLINICAL RELEVANCE: Evaluation of renal size and function may be of benefit for clinical evaluation of feline renal transplant patients, whereas measurement of the resistive index may be of little clinical value. PMID- 10376912 TI - Right atrial compartmentalization using radiofrequency catheter ablation for management of patients with refractory atrial fibrillation. AB - INTRODUCTION: Atrial fibrillation (AF) is often refractory to antiarrhythmic drugs, and patients who are intolerant of AF may require the maze operation for cure. As a less invasive alternative, a catheter-based, right atrial compartmentalization procedure was evaluated. METHODS AND RESULTS: Twelve patients with AF refractory to Class I and III antiarrhythmic drugs were studied. Four linear right atrial radiofrequency ablations were performed, from superior to inferior vena cava in the posterior wall and interatrial septum, anteriorly from the superior vena cava to the tricuspid annulus through the appendage, and across the tricuspid valve-inferior vena cava isthmus. The radiofrequency catheter was dragged along each line three to four times, until the atrial electrogram amplitude decreased by 75% and there was bidirectional conduction block in the tricuspid valve-inferior vena cava isthmus. One complication occurred: sinus node dysfunction requiring a pacemaker. Eight patients were discharged from the hospital on no antiarrhythmic drugs, and four were discharged on previously ineffective antiarrhythmic drugs. Total duration of follow-up was 21.3 +/- 11.2 months. Four patients discharged on previously ineffective antiarrhythmic drugs had no recurrence of AF. One patient discharged off antiarrhythmic drugs had no recurrence of AF. Seven patients discharged off antiarrhythmic drugs had recurrent AF by 12.6 +/- 13.0 months (median 6, range 1 to 39); 3 of these 7 responded to previously ineffective antiarrhythmic drugs without further AF and 4 did not. Thus, 8 of 12 patients (67%) had suppression of AF after ablation on previously ineffective medication or no medication. CONCLUSION: Right atrial compartmentalization may alter the substrate for AF, thus improving the efficacy of previously ineffective antiarrhythmic drugs. Because it is relatively safe, it may be a reasonable adjunctive intervention to maintain sinus rhythm in patients with drug-refractory AF. PMID- 10376913 TI - Intraoperative radiofrequency ablation of chronic atrial fibrillation: a left atrial curative approach by elimination of anatomic "anchor" reentrant circuits. AB - INTRODUCTION: The percutaneous approach to radiofrequency (RF) catheter ablation for curative treatment of atrial fibrillation (AF) is an investigational technique, and the optimal composition of lesion lines is unknown. We tested an intraoperative RF ablation concept with elimination of left atrial anatomic "anchor" reentrant circuits. METHODS AND RESULTS: In 12 patients with an indication for valve surgery and chronic AF, a right atrial-transseptal approach was chosen for access to the left atrium. AF had been present for 4.3 +/- 3.9 years; the left atria measured 56 +/- 7 mm. Under direct vision, contiguous lesion lines were placed endocardially with temperature-guided RF energy applications for treatment of AF with a specially designed probe. The lesion lines were placed between the mitral annulus and the left lower pulmonary vein, further to the left upper pulmonary vein, from there to the right upper pulmonary vein, and finally to the right lower pulmonary vein. The antiarrhythmic ablation procedure lasted 19 +/- 4 minutes. One patient died postoperatively of low cardiac output. During follow-up of 11 +/- 6 months, chronic AF was ablated successfully in 9 of 11 patients (82%). Six patients were in stable sinus rhythm or intermittent pacemaker rhythm, and three patients were in sinus rhythm with intermittent atypical atrial flutter. CONCLUSIONS: Intraoperative RF energy application for induction of contiguous lesion lines is feasible. Elimination of anatomically defined "anchor" reentrant circuits within the left atrium prevented chronic AF in > 80% of the patients treated. Intraoperative validation of lesion line concepts for curative treatment of AF may be transferred to percutaneous ablation techniques. PMID- 10376914 TI - Intraoperative computerized mapping of ventricular tachycardia: differences between anterior and inferior myocardial infarctions. AB - INTRODUCTION: Although direct ventricular tachycardia (VT) surgery has been shown to be effective for treatment of inferior myocardial infarction (MI), the differences in the arrhythmia substrates compared to anterior MI have not been systematically delineated. We sought to compare operative procedures and VT substrates between anterior and inferior MI locations. METHODS AND RESULTS: Computerized mapping was performed in 30 patients with a 128-electrode system using epicardial sock and transatrial left ventricular endocardial balloon arrays, followed by combined endocardial resection and cryoablation. At surgery, there were 51 and 34 different VTs in 18 patients with anterior MI and 12 patients with inferior MI, respectively. The proportion of aneurysms was lower in inferior MI (25% vs 78%, P = 0.008). Total activation times accounted for 65% +/- 23% and 50% +/- 22% of the VT cycle length in anterior and inferior infarcts, respectively (P = 0.005). Complete superficial reentry was identified in 12 VTs related to anterior infarcts and in only two VTs associated with inferior infarction (P = 0.038). Involvement of papillary muscles occurred in two patients with inferior MI. Patients with inferior infarcts received more cryolesions and required epicardial cryolesions or mitral valve replacement more frequently, and their operative mortality was greater (2/12 vs 0/18). Noninducibility rate (89.3%) and 2-year survival (76% +/- 8%) did not differ according to infarct location. CONCLUSION: VT associated with inferior MI can be ablated successfully; however, the substrate is more complex, with frequent participation of intramural layers rendering the ablative procedure more difficult. PMID- 10376915 TI - Verification of linear lesions using a noncontact multielectrode array catheter versus conventional contact mapping techniques. AB - INTRODUCTION: Creation of linear lesions is an established ablation goal. Verification of complete conduction block at the ablation line is required to determine ablation success. Conventional mapping techniques are sequential endocardial activation mapping and documentation of double potentials. Recently, a noncontact multielectrode array catheter was developed that allows instantaneous three-dimensional mapping by simultaneous reconstruction of > 3,000 electrograms. In this study, we prospectively compared the accuracy of noncontact mapping to identify discontinuities in linear lesions and to verify a conduction block with that of conventional mapping techniques. METHODS AND RESULTS: In 12 patients with atrial flutter, radiofrequency pulses were applied between the tricuspid annulus and either the inferior vena cava or the eustachian ridge. Following each application, pulse propagation at the ablation line was determined during pacing by conventional mapping techniques. The findings were compared to high-density isopotential mapping using the noncontact multielectrode array catheter. It was found that noncontact mapping reliably distinguished conduction delays from a conduction block as defined by contact mapping. In addition, noncontact mapping instantaneously identified the area where a discontinuity in the line of block was present. In these patients, complete conduction block was achieved by radiofrequency pulses guided by the noncontact mapping system. CONCLUSION: Noncontact mapping is highly accurate in distinguishing conduction delays from a complete conduction block. By providing an instantaneous high density propagation vector at all sites along the ablation line, three dimensional isopotential mapping is helpful in localizing discontinuities of linear lesions and, thus, may facilitate the creation of a complete conduction block. PMID- 10376916 TI - Target temperatures of 48 degrees C versus 60 degrees C during slow pathway ablation: a randomized comparison. AB - INTRODUCTION: The relationship between temperature at the electrode-tissue interface and the loss of AV and ventriculoatrial (VA) conduction is not established, and the optimal target temperature for the slow pathway approach to radiofrequency ablation of AV nodal reentrant tachycardia (AVNRT) is unknown. Therefore, the purpose of this study was to compare target temperatures of 48 degrees C and 60 degrees C during the slow pathway approach to ablation of AVNRT. METHODS AND RESULTS: The study included 138 patients undergoing ablation for AVNRT. Patients undergoing slow pathway ablation using closed-loop temperature monitoring were randomly assigned to a target temperature of either 48 degrees C or 60 degrees C. The primary success rates were 76% in the patients assigned to 48 degrees C and 100% in the patients assigned to 60 degrees C (P < 0.01). The ablation procedure duration (33 +/- 31 min vs 26 +/- 28 min; P = 0.2), fluoroscopic time (25 +/- 15 min vs 24 +/- 16 min; P = 0.5), and mean number of applications (9.3 +/- 6.5 vs 7.8 +/- 8.1; P = 0.3) were similar in patients assigned to 48 degrees and 60 degrees C, respectively. The mean temperature (46.1 degrees +/- 24.8 degrees C vs 48.7 +/- 3.2 degrees C; P < 0.01), the temperature associated with junctional ectopy (48.1 degrees +/- 2.0 degrees C vs 53.5 degrees +/- 3.5 degrees C, P < 0.0001), and the frequency of VA block during junctional ectopy (24.6% vs 37.2%; P < 0.0001) were less in the patients assigned to 48 degrees C compared to 60 degrees C. The frequency of transient or permanent AV block was similar in each group (2.8% vs 3.6%; P = 0.2). In the 60 degrees C group, only 12% of applications achieved an electrode temperature of 60 degrees C. During follow-up of 9.9 +/- 4.2 months, there was one recurrence of AVNRT in the 48 degrees C group and none in the 60 degrees C group. CONCLUSIONS: Compared to 48 degrees C, a target temperature of 60 degrees C during radiofrequency slow pathway ablation is associated with a higher primary success rate and a higher incidence of VA block during junctional ectopy induced by the radiofrequency energy. AV block is not more common with the higher target temperature, but only if VA conduction is aggressively monitored during applications of radiofrequency energy. PMID- 10376917 TI - The valve of Vieussens: an important cause of difficulty in advancing catheters into the cardiac veins. AB - INTRODUCTION: The coronary sinus and cardiac veins are useful conduits for the passage of electrode catheters for mapping the origin of cardiac arrhythmias. However, sometimes it is difficult to advance catheters an adequate distance into the cardiac veins. The aim of this study was to determine the reasons for this. METHODS AND RESULTS: In 50 cadaveric hearts, a deflectable 7-French electrode catheter was passed from the right atrium into the coronary sinus and advanced to the anterior interventricular portion of the great cardiac vein (GCV). Causes of obstruction were determined. The catheter was obstructed by the valve of Vieussens in 23 of 50 hearts (46%). Once the valve was negotiated, obstruction was caused by an acute bend in the GCV in 28 of 50 hearts (56%). Clinical studies were undertaken in 10 patients in whom electrode catheters could not be advanced as far as required. Using contrast venography, the most frequent cause of obstruction was determined to be the valve of Vieussens in 8 of 10 cases (80%). An acute bend in the GCV caused obstruction in 2 cases (20%). CONCLUSIONS: The valve of Vieussens is a frequent cause of obstruction to passage of a catheter in postmortem and in vivo studies. An acute bend in the vein, with or without lodgment in a tributary, is the other common cause. In adults, venous luminal diameter is not a cause of obstruction to the passage of a 7-French catheter in the coronary sinus or proximal GCV. PMID- 10376918 TI - Muscarinic receptor stimulation with edrophonium hydrochloride does not elevate ventricular fibrillation thresholds in humans. AB - INTRODUCTION: Although decreased vagal tone, as measured by heart rate variability is a risk factor for ventricular fibrillation (VF) and sudden cardiac death, it is unknown whether increasing vagal tone has an antiarrhythmic effect. The purpose of this study was to determine whether edrophonium hydrochloride (HCI), a vagomimetic agent, increases VF threshold. METHODS AND RESULTS: Twenty eight consecutive patients with previously implanted defibrillators had two inductions of VF by monophasic direct-current shocks delivered at 10 to 30 msec after the T wave peak, escalating energies (0.4, 1, then 3 J) until VF was induced. If VF was not induced, this protocol was repeated at the T wave peak and then at 10 to 30 msec before the T wave until VF was induced. Patients were randomized to receive edrophonium HCl (12 to 18 mg) or no drug before repeating the protocol for the second VF induction. The mean sinus cycle length increased from 782 to 872 msec in the group receiving edrophonium HCI (P = 0.006 ). In the control group, the mean sinus cycle length remained unchanged (838 vs 858 msec). The mean energy to induce VF, coupling interval relative to the T wave, and the number of attempts to induce VF were not different between VF induction attempts 1 and 2, and they were not different between the group receiving edrophonium HCl and the control group. CONCLUSION: In a sedated patient population with implantable defibrillators, edrophonium HCI infusion prolongs sinus cycle length but does not change inducibility of VF using T wave shocks. PMID- 10376919 TI - Functional effects of mutations in KvLQT1 that cause long QT syndrome. AB - INTRODUCTION: The long QT syndrome (LQT) is caused by mutations in genes encoding ion channels that modulate the duration of ventricular action potentials. One of these genes, KVLQT1, encodes an alpha subunit that coassembles with another subunit, hminK, to form the cardiac slow delayed rectifier (I(Ks)) K+ channel. METHODS AND RESULTS: The functional effects of seven mutations in KVLQT1 were assessed using two-microelectrode voltage clamp and the Xenopus oocyte expression system. Most mutations in KVLQT1 caused loss of function when expressed alone. Oocytes were also injected with equal amounts of wild-type (WT) KVLQT1 and mutant KVLQT1 cRNA (with or without coinjection of hminK) and the resulting currents compared to currents induced by WT KvLQT1 alone. A341V, R190Q, or G189R KVLQT1 subunits did not affect expression of WT KvLQT1. The other mutations in KVLQT1 caused a variable degree of dominant-negative suppression of I(Ks). The order of potency for this effect was G345E > G306R = V254M > A341E. CONCLUSIONS: LQT1 associated mutations in KVLQT1 caused a spectrum of dysfunction in I(Ks) and KvLQT1 channels. The degree of I(Ks) dysfunction did not correlate with the QTc interval or the presence of symptoms in the respective gene carriers. In contrast to previous reports, we found that loss of function mutations are not exclusive to recessively inherited LQT. PMID- 10376920 TI - Gene expression of the natriuretic peptide system in atrial tissue of patients with paroxysmal and persistent atrial fibrillation. AB - INTRODUCTION: Circulating cardiac natriuretic peptides play an important role in maintaining volume homeostasis, especially during conditions affecting hemodynamics. During atrial fibrillation (AF), levels of plasma atrial natriuretic peptide (ANP) becomes elevated. The aim of this study was to gather information about gene expression of the natriuretic peptide system on the atrial level in patients with AF. METHODS AND RESULTS: Right atrial appendages of 36 patients with either paroxysmal or persistent AF were compared with 36 case matched controls in sinus rhythm for mRNA expression of pro- atrial natriuretic peptide (pro-ANP), pro-brain natriuretic peptide (pro-BNP), and their natriuretic peptide receptor type-A (NPR-A). We investigated patients without (n = 36) and with (n = 36) valvular disease. Persistent AF was associated with higher mRNA expression of pro-BNP (+66%, P = 0.04, in patients without valvular disease, and +69%, P < 0.01, in patients with valvular disease) and lower mRNA expression of NPR-A (-58%, P = 0.02, in patients without valvular disease, and -62 %, P < 0.01, in patients with valvular disease). The mRNA content of pro-ANP was only increased in patients with valvular disease (+12%, P = 0.03). No changes were observed in patients with paroxysmal AF. CONCLUSION: This study demonstrates that persistent, but not paroxysmal, AF induces alterations in gene expression of pro BNP and NPR-A on the atrial level. Although AF generally is associated with an increase of plasma ANP level, a change in mRNA content of pro-ANP is only observed in the presence of concomitant valvular disease and is of minor magnitude. PMID- 10376921 TI - Block of HERG potassium channels by the antihistamine astemizole and its metabolites desmethylastemizole and norastemizole. AB - INTRODUCTION: The selective H1-receptor antagonist astemizole (Hismanal) causes acquired long QT syndrome. Astemizole blocks the rapidly activating delayed rectifier K+ current I(Kr) and the human ether-a go-go-related gene (HERG) K+ channels that underlie it. Astemizole also is rapidly metabolized. The principal metabolite is desmethylastemizole, which retains H1-receptor antagonist properties, has a long elimination time of 9 to 13 days, and its steady-state serum concentration exceeds that of astemizole by more than 30-fold. A second metabolite is norastemizole, which appears in serum in low concentrations following astemizole ingestion and has undergone development as a new antihistamine drug. Our objective in the present work was to study the effects of desmethylastemizole, norastemizole, and astemizole on HERG K+ channels. METHODS AND RESULTS: HERG channels were expressed in a mammalian (HEK 293) cell line and studied using the patch clamp technique. Desmethylastemizole and astemizole blocked HERG current with similar concentration dependence (half-maximal block of 1.0 and 0.9 nM, respectively) and block was use dependent. Norastemizole also blocked HERG current; however, block was incomplete and required higher drug concentrations (half-maximal block of 27.7 nM). CONCLUSIONS: Desmethylastemizole and astemizole cause equipotent block of HERG channels, and these are among the most potent HERG channel antagonists yet studied. Because desmethylastemizole becomes the dominant compound in serum, these findings support the postulate that it becomes the principal cause of long QT syndrome observed in patients following astemizole ingestion. Norastemizole block of HERG channels is weaker; thus, the risk of producing ventricular arrhythmias may be lower. These findings underscore the potential roles of some H1-receptor antagonist metabolites as K+ channel antagonists. PMID- 10376922 TI - Characteristics of I(K) and its response to quinidine in experimental healed myocardial infarction. AB - INTRODUCTION: Mechanisms and drug treatment of serious ventricular arrhythmias in patients with healed myocardial infarction (HMI) are incompletely understood, in part because the electrophysiology and pharmacology of myocytes from noninfarcted regions of HMI hearts are not well characterized. METHODS AND RESULTS: We studied the delayed rectifier potassium current (I(K)) and quinidine responsiveness of single left ventricular subendocardial myocytes isolated from the region remote to the border zone of healed infarct myocardium (4 to 6 mm from scar edge) in cat hearts 2 months after coronary artery occlusion. Subendocardial cells isolated from corresponding regions of normal cat hearts provided controls. I(K) activation and tail currents were recorded using whole cell, voltage clamp techniques. Membrane capacitance of cells remote to HMI (187 +/- 7 pF) was significantly greater than normal (155 +/- 6 pF; P < 0.001). Action potential durations (APDs) recorded from myocytes in remote regions were prolonged (APD90 = 247 +/- 10 msec) compared to normal (214 +/- 11 msec; P < 0.05). Quinidine (1 microM) significantly prolonged APD90 in normal cells but not in remote cells. Density of I(K) (tail current) was significantly decreased in remote cells (3.1 +/- 0.3 pA/pF) compared to normal (3.9 +/- 0.3 pA/pF; P < 0.05), and voltage dependent activation of I(K) was shifted in the positive direction. Quinidine had significantly less incremental blocking effect on I(K) already blunted by regional hypertrophy compared to its effect on normal cells in remote cells. IC50 shifted to 0.95 microM in remote cells compared with 0.50 microM in normal cells. CONCLUSION: Cells in noninfarct region remote from the scar are hypertrophied and display altered electrophysiology. Their reduced I(K) responsiveness to quinidine may explain, in part, failure of quinidine to prolong APD in such cells. Moreover, dispersion of repolarization may be decreased by the effect of quinidine on normal cells. PMID- 10376923 TI - Rapid delayed K+ current and quinidine sensitivity are reduced in healed myocardial infarction. PMID- 10376924 TI - Ablation of ventricular tachycardia with a saline-cooled radiofrequency catheter: anatomic and histologic characteristics of the lesions in humans. AB - INTRODUCTION: In animal models, active cooling of the electrode during radiofrequency (RF) ablation allows creation of larger lesions, presumably by increasing the power that can be delivered without coagulum formation. These RF lesions have not been characterized in human myocardium in regions of infarction and scarring. METHODS AND RESULTS: Cooled-tip RF catheter ablation of ventricular tachycardias (VTs) was performed in two patients who had severe congestive heart failure and subsequently underwent cardiac transplantation. The first patient had four different monomorphic VTs. RF applications along the inferoseptal margin of a scarred region abolished all inducible VTs. The second patient had sarcoidosis involving the myocardium and four different inducible VTs. RF current applied at an inferobasal VT exit and at the right and left septa failed to abolish the VTs. The explanted hearts were examined at the time of cardiac transplantation 18 and 21 days later, respectively. Lesions extended to depths up to 7 mm, reaching clusters of myocardial cells deep to regions of fibrosis. Microscopically, the ablation sites contained coagulation necrosis with hemorrhage, surrounded by a rim of granulation tissue. CONCLUSION: Saline-irrigated RF catheter ablation produces relatively large lesions capable of penetrating deep into scarred myocardium. PMID- 10376925 TI - The role of atrial electrical remodeling in the progression of focal atrial ectopy to persistent atrial fibrillation. AB - Although atrial fibrillation- (AF) induced changes in atrial refractoriness (atrial electrical remodeling) have been demonstrated in a number of different animal models, the clinical significance of this process is unknown. We describe a patient in whom there has been documented progression of atrial ectopy to persistent AF accompanied by evidence of atrial electrical remodeling, with reversal of remodeling following successful ablation of the focal source of AF. A second patient with focal AF, but with a "nonfocal" appearance on the ECG, is also described. These cases illustrate: (1) the possibility that a significant proportion of younger patients with idiopathic persistent AF may well have a focal source as the underlying abnormality; and (2) atrial electrical remodeling reverses following ablation of the underlying source. PMID- 10376926 TI - Software error resulting in malfunction of an implantable cardioverter defibrillator. AB - Loss of ventricular output resulting from an unexpected software error in a dual chamber implantable cardioverter defibrillator (ICD) is reported. A 70-year-old man with a dual chamber ICD implanted for a history of cardiac arrest and infra Hisian block presented with acute onset of dizziness. He was found to have loss of ventricular output due to an internal software problem. The problem was corrected by software reprogramming via the programmer. This malfunction exemplifies the potential ability to correct current-generation ICD software problems noninvasively, thus avoiding the need for replacement. PMID- 10376927 TI - Management of vasovagal syncope. AB - Vasovagal syncope is a common disorder of autonomic cardiovascular regulation that can be very disabling and result in a significant level of psychosocial and physical limitations. The optimal approach to treatment of patients with vasovagal syncope remains uncertain. Although many different types of treatment have been proposed and appear effective based largely on small nonrandomized studies and clinical series, there is a remarkable absence of data from large prospective clinical trials. However, based on currently available data, the pharmacologic agents most likely to be effective in the treatment of patients with vasovagal syncope include beta blockers, fludrocortisone, and alpha adrenergic agonists. In this article, we provide a summary of the various therapeutic options that have been proposed for vasovagal syncope and review the clinical studies that form the basis of present therapy for this relatively common entity. PMID- 10376928 TI - Anisotropy of cardiac tissue: a major determinant of conduction? PMID- 10376929 TI - A paucity of P waves. PMID- 10376930 TI - Ventricular tachycardia as a consequence of a suicidal stabbing injury to the heart. PMID- 10376931 TI - Plasminogen activators and matrix metalloproteases, mediators of extracellular proteolysis in inflammatory demyelination of the central nervous system. AB - The role of extracellular proteolysis in inflammatory demyelination, originally hypothesized as a mechanism for myelin degradation, is increasingly recognized as a pathogenetic step and as a target for therapy in human demyelinating disease. The activation of ubiquitous plasminogen by urokinase (u-PA) and tissue-type plasminogen activator (t-PA), which is associated with various neuropathologies, including multiple sclerosis (MS), is the key initiator of the activation cascade of the four classes of matrix metalloproteinases (MMPs): collagenases, stromelysins, membrane-type metalloproteinases and gelatinases. Spatiotemporal protein and mRNA expression of gelatinase B (MMP-9) and matrilysin (MMP-7) have been documented respectively in MS lesions and in the central nervous system (CNS) of animals developing experimental autoimmune encephalomyelitis (EAE). A close interaction between disease-promoting cytokines and extracellularly acting proteases is deduced from in vitro experiments. Cytokines regulate the balance between the proteases and their respective specific inhibitors at the transcriptional level, while proteolysis is a reciprocal mechanism to enhance (by activation) or downmodulate (by degradation) the specific activities of cytokines. In acute inflammation the contribution of chemokines is hierarchically organised, interleukin-8 (IL-8) and related CXC-chemokines inducing a rapid influx of neutrophils in the acute lesions and an instantaneous exocytosis of gelatinase B granules. This results in sudden and extensive damage to the CNS. In chronic disease involving autoimmune processes CC-chemokines that act mainly on mononuclear cell types appear to be more strictly regulated. As MMPs modify matrix components, promoting extravasation of lymphocytes and monocytes/macrophages and have the potential to generate encephalitogenic peptides from myelin basic protein, novel treatments for demyelinating diseases may be predicted by specific inhibition of these enzymes. Here we review plasminogen activators and the MMP family, in the context of their role in CNS inflammation and demyelination and highlight studies in which intervention in these protease cascades are and may be used to treat demyelinating diseases. PMID- 10376933 TI - HIV-1 tat protein induces the production of interleukin-8 by human brain-derived endothelial cells. AB - This study focused on the role of the HIV-derived viral protein, tat, in activating central nervous system (CNS)-derived endothelial cells (EC) to produce interleukin-8 (IL-8), a stimulator and chemoattractant for neutrophils and lymphocytes. Human CNS-EC treated with tat (100 ng/ml) demonstrated a 2 to 3 fold upregulation in IL-8 mRNA and protein. Tumor necrosis factor-alpha (TNF) and tat were found to act additively in upregulating IL-8 production. In contrast, transforming growth factor beta (TGF beta), appeared to down modulate tat-induced IL-8 production. These data suggest that extracellular tat, especially in the presence of TNF, may be responsible for the local production of IL-8. PMID- 10376934 TI - Requirement of PI3-kinase activity for the nuclear transport of prolactin in cloned murine T lymphocytes. AB - The murine T-cell clone, L2, requires both IL2 and PRL to proliferate. Proliferation and selected IL2-driven gene expression are blocked by treatment with rapamycin. Since prolactin translocation to the nucleus is IL2 dependent and required for proliferation, experiments were performed to identify activation pathways that might be involved in nuclear transport and proliferation. L2 cells were stimulated with IL2 in the presence and absence of the mTOR inhibitor rapamycin, PI3-kinase inhibitors (wortmannin, LY294002), the p38 MAP kinase inhibitor SB203580 and the vitamin D analog calcipotriol. The immunosuppressant rapamycin markedly inhibited IL2-induced proliferation and prolactin translocation to the nucleus. Similarly, wortmannin and LY294002 inhibited IL2 induced proliferation and markedly decreased the amount of prolactin transported to the nucleus. SB203580 and calcipotriol partially inhibited IL2-induced proliferation but had no effect on prolactin translocation. None of the inhibitors affected Lucifer Yellow uptake indicating that rapamycin, wortmannin and LY294002 did not inhibit early endosomal formation but rather worked to inhibit prolactin translocation at a later point in the retrograde transport pathway. PMID- 10376932 TI - High affinity single-chain Fv antibody fragments protecting the human nicotinic acetylcholine receptor. AB - Univalent antibody fragments directed against the main immunogenic region (MIR) of the human acetylcholine receptor (AChR) are capable of protecting the AChR against loss induced by antibodies from myasthenia gravis (MG) patients. Our aim was to construct single-chain Fv (scFv) antibody fragments as a first step towards the production of therapeutic protecting molecules, from two high affinity anti-MIR monoclonal antibodies (mAb 192 and mAb 195). During the construction of scFv192 fragment, two light chains co-secreted from the hybridoma mAb192 were identified. N-terminal amino acid and cDNA sequence analysis showed that one of the two light chains corresponded to the antigen binding molecule while the other originated from the non-secreting myeloma S194/5.XXO.BU.1 which was used in the production of the hybridoma. Functional scFv 192 and 195 fragments were constructed, expressed in Escherichia coli and affinity purified. The binding affinities of scFv192 and scFv195 (K(D) = 0.6 and 0.8 nM for human AChR) were two orders of magnitude higher than that of the earlier constructed scFv198. The scFv192 almost completely protected human AChR against binding of intact anti-MIR mAbs. Human AChR was also very efficiently protected (74-85%) by the scFv192 against binding of autoantibodies from MG sera with high anti-alpha subunit antibody fractions. These scFvs are good candidates for protection of MG patients after appropriate genetic modifications. PMID- 10376936 TI - Schmidt-Lanterman's incisures--the principal target of autoimmune attack in demyelinating Guillain-Barre syndrome? AB - We used immunocytochemical staining of peripheral (trigeminal) nerve to screen sera of patients with Guillain-Barre syndrome (GBS) for the presence of autoantibodies, using sera from patients with other neurological diseases and healthy volunteers as controls. Most sera mildly reacted with axons, myelin sheaths, or sensory neurons without correlation to a specific disease. A characteristic staining, however, was found in 23 demyelinating cases (89%) out of 26 investigated GBS sera. With these sera, dark, oval and often paired small blobs were observed throughout the sections. A similar picture was rarely observed with sera from patients with other disorders or healthy controls. Using immunocytochemical marker proteins and high light microscopic resolution, the blobs were identified as Schmidt-Lanterman's incisures (SLIs). Further investigations will be necessary to identify the corresponding antigen and to answer the question, whether these antibodies represent an epiphenomenon or play a role in the causative mechanism of the disease. PMID- 10376935 TI - Delta opioid receptors expressed by stably transfected jurkat cells signal through the map kinase pathway in a ras-independent manner. AB - Delta opioid receptors (DOR) are G-protein coupled 7-transmembrane receptors (GPCR), expressed by thymic and splenic T cells, that modulate interleukin (IL)-2 production and proliferation in response to concanavalin A or crosslinking the TCR. Mitogen-activated protein kinases (MAPKs) are involved in mediating intracellular responses to TCR crosslinking. In addition, MAPKs can be activated by signaling cascades that are initiated by the release of G-proteins from GPCRs. To determine whether DORs expressed by T cells signal through the MAPKs, extracellular-regulated kinases (ERKs) 1 and 2, two delta opioid peptides, deltorphin and [D-Ala2,D-Leu5]-enkephalin (DADLE), were studied in Jurkat cells that had been stably transfected with DOR (DOR-Ju.1). These peptides rapidly and dose-dependently induced ERK phosphorylation; pretreatment with naltrindole (NTI), a selective DOR antagonist, abolished this. Pertussis toxin (PTX) also inhibited phosphorylation, indicating the involvement of the Gi/o proteins. Herbimycin A, a protein tyrosine kinase (PTK) inhibitor, reduced the DADLE induced ERK phosphorylation by 68%. ERK phosphorylation was inhibited by Bisindolylmaleimide 1 (GF109203X), an inhibitor of PKC, and by pretreatment with PMA prior to DADLE. A GTP/GDP exchange assay was used to assess the potential role of Ras in the pathway leading to ERK phosphorylation; DADLE failed to stimulate GTP/GDP exchange in comparison to PMA. Additional studies showed that DADLE stimulated an increase in cfos mRNA; this was reduced by the inhibitor of MAPK/ERK kinase (MEK), PD98059. Therefore, in DOR-Ju.1 cells, DOR agonists stimulate ERK phosphorylation in a Ras independent and PKC-dependent manner; PTKs appear to be involved. MAPKs mediate the increase in cfos mRNA induced by DOR agonists. PMID- 10376937 TI - Expression of granulocyte colony-stimulating factor in recurrent glial tumors is inversely correlated with tumor progression. AB - We have previously reported that in vivo-expression of granulocyte colony stimulating factor (G-CSF) is a characteristic feature of reactive and neoplastic astrocytes. The aim of the present study was to analyze the expression of G-CSF protein in primary and recurrent astroglial, oligodendroglial and mixed glial differentiated tumors. G-CSF expression was present in all GFAP-positive tumors excepting glioblastomas. G-CSF expression was significantly reduced in recurrent tumors more dedifferentiated than their primary counterparts. G-CSF expression was absent in all GFAP-negative tumors such as oligodendrogliomas. Our results demonstrate that G-CSF is a highly sensitive differentiation marker of neoplastic astrocytes, which is lost during tumor progression. PMID- 10376938 TI - Comparison of C1q-receptors on rat microglia and peritoneal macrophages. AB - A comparison of the expression and ligand specificity of the C1q (first complement component) receptor on rat microglia and peritoneal macrophages was made. This revealed that radiolabelled C1q was competed from the peritoneal macrophages with intact C1q, and additively displaced by calf-skin collagen and purified C1q globular heads, suggesting the presence of at least two receptors. This was in contrast to microglia, where radiolabelled C1q was displaced with intact C1q and to a modest degree with collagen, but not with globular heads. Taken together, this implies that under these conditions, peritoneal macrophages and microglia both express a C1q receptor which binds to the collagen-like region, and that peritoneal macrophages additionally express a molecule which binds to the globular head of C1q. Analysis of the ligand bound by these cells reflected the differences observed in the competitive binding experiments, with the novel identification of naturally-occurring peptides from the globular head of C1q bound to the peritoneal macrophages, but not the microglia. PMID- 10376940 TI - Effect of rhTNF-alpha injection into rat sciatic nerve. AB - To assess whether TNF-alpha causes inflammatory demyelination or axonal degeneration, we injected into rat sciatic nerve saline, 100 U and 1000 U of rhTNF-alpha and studied the electrophysiological and pathological effects. At day 1 electrophysiology showed a slight reduction of proximal compound muscle action potential amplitude and pathology showed edema, inflammatory infiltration of vessel walls and endoneurium only in nerves injected with 1000 U of rhTNF-alpha. At day 5, there was no demyelination and a percentage of degenerated fibers similar in the three groups. To study the blood-nerve barrier, fluorescein isothiocyanate-labelled albumin was given intravenously after intraneural injection. The nerves injected with 1000 U rhTNF-alpha showed a leakage of the tracer in the endoneurium. TNF-alpha does not appear, at the doses used, to have myelinotoxic or axonopathic properties. The electrophysiological effect at day 1 may be due to mechanical compression of nerve fibers as a result of the blood nerve barrier damage with consequent endoneurial edema. PMID- 10376939 TI - Markers in the promoter region of interleukin-10 (IL-10) gene in myasthenia gravis: implications of diverse effects of IL-10 in the pathogenesis of the disease. AB - Interleukin-10 (IL-10) is an important pleiotropic cytokine with both anti inflammatory and B lymphocyte stimulating functions. The expression of IL-10 is tightly controlled. A bi-allelic polymorphism and 2 CA repeat microsatellites located in the promoter region of IL-10 gene were analysed in Swedish myasthenia gravis (MG) patients and ethnically matched healthy individuals. The prevalence of a 'high secretor' phenotype of IL-10 (IL-10 1 G/G) was higher in IL-1beta TaqI polymorphism allele 2 positive healthy individuals ('high secretor' phenotype for IL-1beta) than in healthy individuals negative for this allele. No such balance was found in MG patients. In one of the microsatellites, IL10.R, allele 112 was associated with patients having normal thymic histology. No relation of IL10.R allele 112 to proinflammatory cytokine gene polymorphisms and serum IgG, IgM and autoantibodies against nicotinic acetylcholine receptor (nAchR-Ab) was found. In another microsatellite, IL10.G, allele 134 was associated with patients having higher level of nAchR-Ab in their circulation. Our results demonstrated novel genetic markers within IL-10 gene indicating different mechanisms for IL-10 involved in the disease. PMID- 10376941 TI - Identification and characterization of immunoreactive calcitonin gene-related peptide from lymphocytes of the rat. AB - There is accumulating evidence that the immune system can produce neuropeptides. In the light of these facts, we obtained direct evidences to prove that T lymphocytes also synthesize and secrete calcitonin gene-related peptide (CGRP), a neuropeptide localized within primary sensory nerves. By using CGRP specific RIA, CGRP-like immunoreactivity (LI) was found in the extracts of rat lymphocytes from thymus and mesenteric lymph node. The intracellular concentration of lymphocyte derived CGRP-LI of rat thymus and mesenteric lymph node was 745+/-39 and 447+/-33 fg/10(6) cells, respectively. CGRP-LI in lymphocytes was shown to co-elute with synthetic rat CGRP and sensory neuron-derived CGRP by reverse-phase HPLC. In addition, the CGRP-LI located in the T lymphocytes was also shown by immunocytochemical method examined by electron microscopy. The CGRP mRNA detected by RT-PCR was also present in these lymphocytes and was also identified to be the same one in sensory neurons. These data suggest that CGRP is synthesized and secreted in T lymphocytes of both thymus and lymph node in the rat, and this is identified to be the same one in neuronal tissue. Lymphocyte-derived CGRP may act in an autocrine/paracrine mode and play an important role in certain physiological and pathophysiological conditions. PMID- 10376942 TI - Early differentiation of thymic dendritic cells in the absence of glucocorticoids. AB - The possible role of glucocorticoids (GCs) in the maturation of thymic dendritic cells (DCs) during early ontogeny was analyzed in the progeny of adrenalectomized pregnant rats (Adx foetuses). This experimental model ensured the lack of GCs until establishment of foetal hypothalamus-pituitary gland-adrenal (HPA) axis, and showed profound modifications of the development of thymus gland. In the absence of maternal GCs, there was a high percentage of DCs, many of them exhibiting a mature phenotype, in the 15-16 day-old Adx foetal thymus, which sharply decreased to reach control values on foetal day 17. On the other hand, the absolute number of DCs of Sham foetal rats increased throughout ontogeny, whereas the high numbers found in 15-16 day-old Adx foetuses significantly diminished in the following days. This process was closely correlated with the thymocyte life span, previously demonstrated, and the early appearance of DCs in the spleen. Our results demonstrate that like for other cell components of rat thymus, DC maturation is accelerated in an early foetal microenvironment devoid of glucocorticoids. PMID- 10376943 TI - Antigen-specific immunosuppression: nasal tolerance to P0 protein peptides for the prevention and treatment of experimental autoimmune neuritis in Lewis rats. AB - Experimental autoimmune neuritis (EAN) is an autoimmune inflammatory demyelinating disease of the peripheral nervous system (PNS), and represents an animal model of the human Guillain-Barre syndrome (GBS). In this study, we report that nasal administration of the neuritogenic peptide 180-199 and of the cryptic peptide 56-71 of the rat neuritogenic P0 protein of peripheral nerve myelin prevents EAN and attenuates ongoing EAN. Both peptides effectively decreased the severity and shortened clinical EAN. Both a prophylactic and a therapeutic approach proved to be beneficial. These effects were associated with T and B cells hyporesponsiveness to the peptide antigens, reflected by downregulated Th1 cell responses (interferon-gamma secretion) and macrophage function, whereas Th2 cell responses (IL-4 secretion) and transforming growth factor-beta mRNA expression were upregulated. PMID- 10376944 TI - B cell-deficient mice have increased susceptibility to HSV-1 encephalomyelitis and mortality. AB - We studied the susceptibility of B cell-deficient mice to encephalomyelitis following intraperitoneal inoculation of HSV-1. B cell-deficient mice developed striking CNS signs including tail atony, clumsy gait and limb paralysis after HSV 1 infection. In addition, B cell-deficient mice had decreased survival (LD50 = 2.2 x 10(7) PFU) compared to control C57BL/6 mice (LD50 = 2.3 x 10(8) PFU). B cell-deficient mice had encephalomyelitis and detectable virus in the brain 7 days post-infection while C57BL/6 mice did not. Passive transfer of hyperimmune sera protected B cell-deficient mice from death, suggesting a role for antibody in susceptibility to HSV-1 encephalomyelitis. PMID- 10376945 TI - Separate precursor cells for macrophages and microglia in mouse brain: immunophenotypic and immunoregulatory properties of the progeny. AB - The brain contains two populations of macrophages: the microglia of brain parenchyma, and the central nervous system (CNS) macrophages located in the perivascular spaces, the leptomeninges and the choroid plexus. The microglia are characterized, in part, by their paucity of major histocompatibility complex (MHC) molecules and lack of constitutive antigen (Ag)-presenting activity for naive CD4+ T-cells. Some CNS macrophages, on the other hand, constitutively express MHC molecules and present Ag to naive CD4+ T-cells. We have reported that mouse brain contains precursor cells that, in the presence of colony-stimulating factor-1, the macrophage growth factor, give rise to clones of cells that differ in their ability to constitutively present Ag to naive CD4+ T cells. Here we report that this population of precursor cells can be separated into two discrete subpopulations based on differences in cell density and that the two cell populations give rise to progeny that differ in their content of cells constitutively expressing MHC class II and CD86 molecules, and the ability to present Ag to naive CD4+ T-cells. A comparison of the level of CD45 staining of the progeny, an indication of a microglial or a CNS macrophage origin, suggests that one population of precursor cells yields immunologically immature microglia and the other CNS macrophages. PMID- 10376947 TI - Social stress, dominance and blood cellular immunity. AB - The impact of chronic social coexistence on distribution and function of blood immune cells was examined in Long Evans rats. At the beginning of a 7 day period of chronic coexistence (confrontation), a wall was removed between two neighboring cages each consisting of a male-female pair. Winner and loser males were classified based on differences in their defensive behavior. On day 2 and 7 of confrontation, losers showed reductions in numbers of blood CD4 and CD8 T cells as well as profound suppression of in vitro NK activity and lymphocyte (LYM) proliferation. Numbers of granulocytes (GRAs) were more than doubled. Winner males showed similar immunological alterations only on day 2 of confrontation. On day 7 most changes were reversed. The persistent changes in loser males may reflect a less favorable state for effective immune response. PMID- 10376948 TI - High levels in serum, but no signs of intrathecal synthesis of anti-sulfatide antibodies in HIV-1 infected individuals with or without central nervous system complications. AB - Myelin degeneration is commonly found in the central nervous system (CNS) of individuals infected with human immunodeficiency virus type 1 (HIV-1), especially in patients with HIV-1-associated dementia. We analysed cerebrospinal fluid (CSF) and serum samples from 25 HIV-1 infected individuals for the presence of antibodies directed against sulfatide, the major acidic glycosphingolipid in myelin. Nine of the patients had CNS complications, including 3 with HIV-1 associated dementia, and 16 had no neurological symptoms. Elevated titres of anti sulfatide antibodies were found in serum from 24/25 HIV-1-infected individuals but in none of them in the CSF. Although the vast majority of HIV-1-infected individuals harbour autoantibodies directed against sulfatide in serum, the lack of detectable intrathecal production indicates that anti-sulfatide antibodies are not a major component in the pathogenesis of CNS myelin damage in HIV-1 infection. PMID- 10376946 TI - Interferon-gamma reduces cyclooxygenase-2-mediated prostaglandin E2 production from primary mouse astrocytes independent of nitric oxide formation. AB - Nitric oxide (NO) and prostaglandins (PGs) modulate inflammatory and immune responses in the central nervous system (CNS). Both NO and PG synthesis have been described in appropriately stimulated astrocytes. In other systems, both positive and negative modulation of cyclooxygenase (COX) activity, hence PG synthesis, have been described by NO. Since interferon (IFN)-gamma is known to upregulate the production of NO from astrocytes, the present study was designed to investigate the effect of IFNgamma on PG production from activated astrocytes and to determine whether this effect is mediated by NO. Astrocytic PG production was induced by exposure of murine cortical cultures to lipopolysaccharide (LPS). This induction was time- and concentration-dependent, and prevented by inhibitors of transcription and translation, as well as the selective COX-2 inhibitor, NS-398. LPS-induced expression of COX-2 mRNA and protein was confirmed by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. Exposure of LPS-treated astrocytes to IFNgamma resulted in a concentration-dependent decrease in PGE2 accumulation which was accompanied by a striking parallel increase in NO formation. However, the NOS inhibitors, N(G) nitro-L-arginine or N6-(1-iminoethyl)-lysine, failed to reverse the IFNgamma mediated diminution of LPS-induced PGE2 production, indicating that the IFN-gamma mediated reduction in COX-2-dependent PGE2 production occurred independent of NO formation. Additional experiments demonstrated that IFN-gamma acted mainly by downregulating the expression of COX-2 protein. Present results indicate that PG and NO synthesis in mouse cortical astrocytes in vitro are under the direct reciprocal control of IFNgamma. PMID- 10376950 TI - Tumour necrosis factor-alpha polymorphism and secretion in myasthenia gravis. AB - The mechanism behind the association between MHC genes and myasthenia gravis (MG) is not fully understood. In the present study we studied the associations with polymorphisms at HLA-DR3, HLA-B8 and TNF-alpha genes in Swedish patients and healthy individuals. The TNF-alpha-308 allele 2 was associated with female patients having disease onset before the age 40 and with thymic hyperplasia. Analysis of strongest associations between MG and alleles close to TNF-alpha indicated that the association of TNF-alpha was possibly stronger than for HLA DR3 and nearly the same as for HLA-B8. Peripheral blood mononuclear cells from patients positive for TNF-alpha -308 allele 2 had higher secretion of TNF-alpha when stimulated by anti-CD3 antibodies. Our results indicate that a subgroup of MG patients who have been previously shown to be associated with MHC genes may have a higher inducible TNF-alpha level in vivo, thus resulting the pathological changes in the thymus and the early onset of MG. PMID- 10376949 TI - Anti-GM2 IgM antibodies: clinical correlates and reactivity with a human neuroblastoma cell line. AB - Anti-GM2 IgM antibodies have been reported in some patients with dysimmune neuropathy or lower motor neuron syndrome, in whom they were often associated with a concomitant reactivity with GM1. To investigate the possible clinical and pathogenetic relevance of these antibodies we measured serum anti-GM2 IgM titers by ELISA in 224 patients with different neuropathies and motor neuron disease and examined their binding to SK-N-SH neuroblastoma cells by indirect immunofluorescence (IIF). High titers of anti-GM2 IgM antibodies were found in eight patients with dysimmune neuropathies including two with multifocal motor neuropathy (MMN), two with purely motor demyelinating neuropathy without conduction block (MN) and four with Guillain-Barre syndrome (GBS). In two MMN patients reactivity with GM2 was associated with anti-GM1 reactivity and in one MN patient with anti-GM1, -GD1a and -GD1b reactivity. All but one patient had a concomitant reactivity with GalNAc-GD1a. Serum IgM from all positive patients intensely stained by IIF the surface of SK-N-SH neuroblastoma cells. This reactivity was blocked by serum pre-incubation with GM2, was not observed with sera from patients without anti-GM2 antibodies including those with high anti-GM1 or other anti-glycolipid antibodies, and correlated with the presence of GM2 in the SK-N-SH neuroblastoma cells. These findings indicate that anti-GM2 antibodies, though infrequent, are strictly associated with dysimmune neuropathies and suggest that SK-N-SH neuroblastoma cells can be a suitable in vitro model to study the functional and biological effects of these antibodies. PMID- 10376951 TI - Involvement of catecholamines in recall of the conditioned NK cell response. AB - The primary goal of the study was to identify the types of catecholamines and the associated receptors which might be involved in the recall of the conditioned NK cell response. Specific catecholamine receptor antagonists were selected to block the conditioned NK cell response at the recall step. The regional contents of dopamine (DA), norepinephrine (NE), and epinephrine were determined in the brain of the conditioned animals by using the high performance liquid chromatography with electrochemical detection (HPLC/ED). Results showed that pre-disruption of the central alpha1-, alpha2-, beta1-, beta2-, D1-, or D2-receptors at the conditioned recall stage, interrupted the conditioned enhancement in NK cell activity. The NE contents at the cerebellum, and DA contents at the striatum and hippocampus, were significantly higher in the brain of the conditioned animals when compared to that of the control animals. These information indicated the possible roles of the central noradrenergic and dopaminergic systems in regulating the recall of the conditioned NK cell response. PMID- 10376952 TI - Construction and characterization of a humanized single chain Fv antibody fragment against the main immunogenic region of the acetylcholine receptor. AB - The single chain Fv fragment of mAb198 (scFv198) directed against the main immunogenic region (MIR) of the nicotinic acetylcholine receptor (AChR), can efficiently protect the AChR in muscle cell cultures against the destructive activity of human myasthenic autoantibodies. Humanization of the scFv198 antibody fragment should prove useful for therapeutic application by reducing its immunogenicity. Framework sequences from human immunoglobulins homologous to the rat scFv198 sequences were selected and a totally synthetic humanized scFv198 antibody fragment was constructed in vitro. Humanized VH and VL domains were synthesized using two overlapping sets of 225 bases long oligonucleotides overlap extension and polymerase chain reaction (PCR), then assembled into a full-length gene by overlap extension of single-stranded DNA (ssDNA) fragments and PCR. The initial humanized antibody fragment had a very low affinity for the AChR. Molecular modeling was then performed and four residues from the framework regions (FR) of the humanized VH domain were selected to be replaced by the corresponding amino acid from the rat sequence. Three mutants were constructed by overlap extension, using PCR. The humanized variant containing replacements at VH residues 27, 29, 30 and 71 showed very good recovery of AChR binding activity; its binding affinities for Torpedo or human AChR (K(D): 8.5 or 323 nM, respectively) being only four times lower than those of the parental scFv198 (K(D): 2 or 80 nM, respectively). This variant was able to protect the human AChR against the binding of anti-MIR mAb and anti-alpha autoantibodies from a myasthenic patient. It was also able to protect AChR against antigenic modulation induced by the anti-MIR mAb198. PMID- 10376953 TI - Induction of experimental autoimmune neuritis in CD4-8-C57BL/6J mice. AB - The C57BL/6J mice strain is known to be reputedly resistant to induction of experimental autoimmune neuritis (EAN), an animal model of Guillain-Barre syndrome in humans. Here we describe the induction of EAN in mice of the C57BL/6J background by transfer into naive syngeneic recipients bovine peripheral nerve myelin (BPM)-primed donor lymph node cells that had been stimulated in vitro with the bovine peripheral nervous system (PNS) myelin P2 protein peptide 57-81 followed by challenge with BPM, Freund's complete adjuvant and pertussis toxin. EAN was more severe, both clinically and histologically, and accompanied by extensive infiltration of inflammatory cells and demyelination in peripheral nerves when examined on day 30 after transfer of primed T cells from CD4- 8- mice into identical naive hosts than after transfer of cells from primed wild type, CD4-/- or CD8-/- mice to corresponding recipient animals. EAN in CD4-8- mice was also associated with elevated numbers of P2 peptide-reactive interferon-y (TFN gamma) secreting cells and alphabeta T cells were present in lymph nodes and spleens. The data suggest that PNS myelin activated T cells from an EAN-resistant mice strain are capable of homing to the PNS. The expanded CD4-8- alphabeta T cells may have helper and effector functions, related to initiation of EAN in the CD4-8- mice. Lack of CD4+ and CD8+ expressing cells does not prevent the initiation of an autoimmune disease. PMID- 10376954 TI - Cloning the antibody response in humans with inflammatory CNS disease: isolation of measles virus-specific antibodies from phage display libraries of a subacute sclerosing panencephalitis brain. AB - We have developed a strategy to identify the disease-relevant antigens in a chronic inflammatory CNS disease exhibiting intrathecally expressed oligoclonal IgG. Using subacute sclerosing panencephalitis (SSPE), a chronic inflammatory measles virus infection of the brain as a model system, we constructed a phage display antibody Fab library from the amplified products of IgG expressed in the brain. Selection of the library against measles virus-infected cell lysates yielded four distinct Fabs which, by ELISA and by immunostaining, reacted specifically with measles virus-infected cells. Three Fabs immunoprecipitated a 72 kDa protein from infected cell cultures corresponding to the measles virus phosphoprotein. The fourth Fab immunoprecipitated and recognized by immunoblotting a 60 kDa protein corresponding to the measles virus nucleoprotein. The results demonstrate that functional antibodies from an inflammatory CNS disease can be expressed in bacteria and used to identify disease-relevant antigens. This approach could be applied to chronic inflammatory CNS diseases of unknown cause such as multiple sclerosis. PMID- 10376955 TI - Platelet activating factor is elevated in cerebral spinal fluid and plasma of patients with relapsing-remitting multiple sclerosis. AB - Platelet-activating factor (PAF) is a phospholipid mediator of inflammation with a wide range of biological activities, including the alteration of barrier function of endothelium. A biological assay combined with high pressure liquid chromatography-tandem mass spectrometry showed that plasma and cerebral spinal fluid (CSF) PAF levels in 20 patients with relapsing/remitting or secondary progressive multiple sclerosis (MS) studied by magnetic resonance imaging (MRI) were significantly higher than in healthy controls (plasma: 3.29+/-4.52 vs. 0.48+/-0.36 ng/ml, p < 0.002; CSF: 4.95+/-6.22 ng/ml vs. 0.01+/-0.04 ng/ml, p < 0.0001). Values were also significantly higher in relapsing/remitting than in secondary progressive (plasma: 5.10+/-4.97 vs. 0.52+/-0.85 ng/ml, p < 0.005; CSF: 8.59+/-6.39 vs. 0.55+/-0.68 ng/ml, p < 0.002). It was also found that both plasma (R2: 0.65) and CSF (R2:0.72) levels were correlated with the MRI number of gadolinium enhancing lesions, which are markers of blood-brain barrier (BBB) injury, whereas their peaks were not correlated with the MRI number of white matter lesions, nor with the expanded disability status score (EDSS) according to Kurtze [Kurtze, J.F., 1983. Rating neurological impairment in multiple sclerosis: an expanded disability scale (EDSS). Neurology 33, 1444-1452]. Both plasma and CSF in patients with relapsing/remitting MS and marked gadolinium enhancement contained the two major molecular species of PAF: 1-0-hexadecyl- (C16:O) and 1-0 octadecyl-sn-glycero-3-phosphocholine (C18:O). The ratio of the two molecular species was different in the two biological fluids, being PAF C18:0 more abundant in CSF and PAF C16:0 in plasma, indicating a different cellular origin of PAF or different enzymatic processing. These findings suggest that PAF is a significant mediator of BBB injury in the early stages of MS, rather than a marker of its progression and severity. PMID- 10376956 TI - Creativity over the life course--a call for a relational perspective. AB - Research into creativity over the life course has a long history of concentrating on individual characteristics thought to be associated with shifting patterns. In large measure creativity has been explained as a consequence of intelligence, cognitive style, personality, imagery, and other endogenous factors. These human capital factors are not sufficient in themselves to explain differential creativity either by age or among diverse groups. In addition to individual-level considerations there is an array of societal and domain specific factors that must be taken into account. A number of these are discussed under the rubric of social capital. Finally a relational perspective is advocated as an interpretative framework and discussed in terms of dialectical life span approaches to the life course. PMID- 10376957 TI - Respect for the elderly in Asia: stability and change. AB - This study analyzes data from seventy-nine focus groups conducted in the Philippines, Singapore, Taiwan, and Thailand. The research examined ways in which respect for the elderly is experienced in these four countries, the extent to which respect has changed over time, and the reasons for changes in respect for the elderly. Using qualitative analysis, five distinct dimensions of respect were identified: gestures and manners, tokens, customs and rituals, asking for advice, and obedience. Focus group discussions indicated that changes have occurred on most of these dimensions of respect. The changes were attributed to variations in family structure and function, education, income, and modernization. These findings are discussed in relation to changing definitions of respect and variations in the way in which respect for the elderly is expressed in Asia. PMID- 10376958 TI - The inner self of the Japanese elderly: a defense against negative stereotypes of aging. AB - Japanese scholars have reported that the Japanese people tend to maintain highly developed outer and inner selves. This study examines how these selves impact on aging. We hypothesized that the Japanese elderly would express more negative attitudes toward old people in general but more positive self-concepts than elders in China and the United States. The results supported this predicted pattern. They suggest an unusual dynamic of aging and self-identity in Japan that can shed light on the role of the self in accepting or rejecting societal stereotypes about aging. PMID- 10376959 TI - Disability and depressive symptoms in the elderly: the effects of instrumental support and its subjective appraisal. AB - This article explores the buffering effect of social support on depressive symptoms in a community sample of elderly with varying levels of disability. Baseline interviews were conducted in respondents' homes. Results show that higher levels of disability are associated with higher levels of depression. Instrumental support and subjective appraisal of the network are associated with depressive symptoms, but instrumental support has a weak positive correlation, while subjective appraisals show a negative relationship. Social support mitigates the depressive effect of disability only when the network's efforts are appraised positively. However, no such relationship is shown for instrumental support. One's perception of the network's helpfulness appears to be more potent than the actual help provided by friends and family. PMID- 10376960 TI - Diminution of 37-kDa laminin binding protein expression reduces tumour formation of murine lung cancer cells. AB - Expression of the 37-kDa laminin binding protein (37LBP/p40), a precursor of the 67-kDa laminin receptor, is well-correlated with the biological aggressiveness of cancer cells. To elucidate the direct role played by 37LBP/p40 in cancer cells, a murine lung cancer cell line T11, the 37LBP/p40 expression of which was remarkably diminished, was established by the introduction of the antisense 37LBP/p40-RNA using a retroviral vector. As a result, the population doubling time of T11 was prolonged (60 h) compared with that of P29, the non-transfected parental cell line (42 h), and TN2, a transfectant with vehicle only (40 h). In vitro studies also showed that T11 cells adhered to immobilized laminin less firmly than P29 cells did. When 5 x 10(5) cells were subcutaneously inoculated into syngenic mice, the mean survival time of T11-recipients (77.0+/-14.8 days) was also significantly prolonged compared with that for P29 (34.8+/-5.5 days) and TN2 (36.7+/-6.1 days) recipients (P < 0.001). The electron-microscopic view of the tumour tissue revealed that T11 cells were loosely apposed and their intercellular space was markedly widened. Some of the T11 cells sporadically degenerated with the infiltration of lymphocytes and neutrophils. These results suggest that the suppressed expression of 37LBP/p40 reduces the capability of lung cancer cell proliferation in vitro and tumour formation in vivo. PMID- 10376961 TI - Regulation and expression of multidrug resistance (MDR) transcripts in the intestinal epithelium. AB - A paucity of information exists on the regulation of gene expression in the undifferentiated intestine. The intestinal epithelium is one of the few normal tissues expressing the multidrug resistance (MDR) genes that confer the multidrug resistant phenotype to a variety of tumours. Expression of mdr1a has been observed in the primitive rat intestinal epithelial cell line, IEC-18. It is hypothesized that characterization of MDR gene expression in IEC-18 cells will provide insight into gene regulation in undifferentiated intestinal cells. A series of hamster mdr1a promoter deletion constructs was studied in IEC-18 and a region with 12-13-fold enhancer activity was identified. This region was shown to function in an orientation- and promoter context-independent manner, specifically in IEC-18 cells. Unexpectedly, Northern probing revealed a greater expression of mdr1b than mdr1a in IEC-18 cells. A quantitative reverse transcription polymerase chain reaction assay was used to compare the relative expression of MDR genes in IEC cells, fetal intestine, and in the undifferentiated and differentiated components of adult intestinal epithelium. MDR transcript levels in IEC cells were found to resemble those of fetal intestine and small intestinal crypts, where a conversion from mixed mdr1a/mdr1b to predominantly mdr1a expression occurs as cells mature. This work describes two contributions to the field of gene regulation in the undifferentiated intestine--first, the initial characterization of a putative mdr1a enhancer region with specificity for primitive intestinal cells and secondly, the first report of mdr1b detection in the intestine and its expression in primitive cell types. PMID- 10376962 TI - Correlation of neomycin, faecal neutral and acid sterols with colon carcinogenesis in rats. AB - High fat diets have been implicated in incidence of colon cancer both in epidemiological and animal studies. Present investigation deals with the incidence, location and numbers of large and small bowel tumours induced by 1,2 dimethyl hydrazine (DMH) in rats fed high fat diets and neomycin. Neomycin was used to modify the faecal sterol metabolism and the relationship of the high fat diet and faecal neutral and acid sterols to the large bowel tumorigenesis was evaluated. DMH administered rats were fed with (a) 20% safflower oil; (b) 20% safflower oil and neomycin; (c) 20% safflower oil, cholesterol and cholic acid; and (d) 20% safflower oil, cholesterol, cholic acid and neomycin. Neomycin was found to be associated with both increase and decrease of tumour numbers. The faecal sterols lithocholic and deoxycholic acids were found to have no participation, while cholesterol and cholic acid were found to decrease with increase in tumour numbers. However, faecal coprostanol has been found to have a significant positive correlation with tumorigenesis in all dietary groups. Therefore coprostanol might possibly be associated with colon carcinogenesis in DMH-fed rats and cholesterol metabolism in gut appears to be related to the development of tumours. PMID- 10376963 TI - Enhancement of membrane-type 1-matrix metalloproteinase (MT1-MMP) production and sequential activation of progelatinase A on human squamous carcinoma cells co cultured with human dermal fibroblasts. AB - Matrix metalloproteinase 2 (MMP-2)/gelatinase A plays an important role in tumour invasion and metastasis. Since MMP-2 is secreted as an inactive form (proMMP-2) from tumour and neighbouring stroma cells, the activation process is necessary to express the enzymic activity for degradation of extracellular matrix components. We herein reported that the activation of proMMP-2 was induced in human squamous carcinoma cells co-cultured with normal human dermal fibroblasts. When A431 cells were co-cultured with human fibroblasts at various cell ratios, 72-kDa proMMP-2 was converted to a 62-kDa active form through the appearance of a 64-kDa intermediate. The activation of proMMP-2 by co-culture was also observed in other carcinoma cell lines, HSC-4 and SAS, but not in normal human keratinocytes. We characterized by in vitro invasion assay that A431 cells in co-culture preferentially invaded through Matrigel and the increased invasive activity was inhibited by exogenously adding tissue inhibitor of metalloproteinases 2. The augmented proMMP-2 activation by co-culture was achieved by the increase in membrane type 1-MMP (MT1-MMP) production along with that of its mRNA level. The predominant appearance of MT1-MMP was immunologically observed in A431 cells, but not human fibroblasts of the co-culture. Furthermore, epidermal growth factor (EGF) enhanced the co-culture-mediated proMMP-2 activation by increasing the production and gene expression of MT1-MMP, and thereby tumour invasive activity was further augmented. These results suggest that the cell-cell contact between carcinoma cells and normal fibroblasts enhances the production of MT1-MMP followed by sequential activation of proMMP-2 on the tumour cell surface, which may be closely implicated in tumour invasion in vivo. PMID- 10376964 TI - Rapid induction of p21WAF1 but delayed down-regulation of Cdc25A in the TGF-beta induced cell cycle arrest of gastric carcinoma cells. AB - Transforming growth factor-beta (TGF-beta) is a multifunctional polypeptide that inhibits cellular proliferation in most epithelial cells. cdk4 and several cyclin dependent kinase (cdk) inhibitors (p15INK4B, p21WAF1/Cip1 and p27Kip1) have been implicated in the TGF-beta-induced cell cycle arrest. More recently, down regulation of Cdc25A, a cdk activator, was additionally suggested as a mechanism underlying growth inhibition by TGF-beta. The existence of diverse cellular mediators of TGF-beta, however, raises the question of whether their involvement might occur in a redundant manner or coordinately in a certain cell type. Using two TGF-beta-sensitive gastric carcinoma cell lines (SNU-16 and -620), we addressed the contributory roles of several cdk inhibitors, and of cdk4 and Cdc25A, in TGF-beta-induced cell cycle arrest by comparing their temporal expression pattern in response to TGF-beta. Among the cdk inhibitors examined, p21 mRNA was most rapidly (in less than 1 h) and prominently induced by TGF-beta. In contrast, p15 mRNA was more slowly induced than p21 in SNU-620 cells, and not expressed in SNU-16 cells harbouring homozygous deletion of p15. Western blotting results confirmed the rapid increase of p21, while opposite patterns of p27 expression were observed in the two cell lines. The down-regulation of Cdc25A mRNA occurred, but was more delayed than that of p15 or p21. Until G1 arrest was established, changes in the protein levels of both Cdc25A and cdk4 were marginal. Co-immunoprecipitation with anti-cdk4 antibody showed that induced p21 associates with cdk4 and that its kinase activity is reduced by TGF-beta, which kinetically correlates closely with G1 arrest following TGF-beta treatment of both cell lines. These results suggest that in certain human epithelial cells, p21 may play an early role in TGF-beta-induced cell cycle arrest, and its cooperation with other cdk inhibitors is different depending on cell type. Delayed down-regulation of Cdc25A and cdk4 may contribute to cell adaptation to the quiescent state in the two gastric carcinoma cell lines studied. PMID- 10376966 TI - Suppression of telomerase reverse transcriptase (hTERT) expression in differentiated HL-60 cells: regulatory mechanisms. AB - Telomerase activity, associated with cellular immortalization and tumorigenesis, is suppressed during terminal differentiation of HL-60 promyelocytic leukaemic cells. However, it is poorly understood how telomerase activity is regulated in differentiated HL-60 cells. In the present study, we demonstrate that the down regulation of telomerase reverse transcriptase (hTERT) expression, the catalytic subunit, occurs prior to the suppression of telomerase activity in differentiated HL-60 cells. In contrast, the expression of telomerase RNA template (hTR) and telomerase associated protein (TP1) is not reduced. This down-regulation of hTERT expression is achieved through inhibition of gene transcription, in which process new protein synthesis is required. Moreover, the rapid down-regulation of hTERT expression followed by the inhibition of telomerase activity is a specific component of the differentiation programme and not simply a consequence of cell cycle arrest. Serum-deprivation of HL-60 cells causes cell cycle arrest without differentiation and this does not result in a significant reduction in hTERT mRNA levels within the first 24 h. Our findings suggest that hTERT expression is stringently controlled at transcriptional level in HL-60 cells. The downregulation of hTERT expression in the HL-60 cell differentiation model may represent a general regulatory mechanism through which telomerase becomes repressed during development and differentiation of human somatic cells. PMID- 10376965 TI - The two phyto-oestrogens genistein and quercetin exert different effects on oestrogen receptor function. AB - We compared the oestrogenic and anti-oestrogenic properties of the two well-known phyto-oestrogens, genistein and quercetin, on the oestrogen-sensitive breast cancer cell line MCF-7. Genistein exerted a biphasic effect on growth of MCF-7 cells, stimulating at low and inhibiting at high concentrations, whereas quercetin was only growth inhibitory. At doses which did not inhibit cell growth, respectively 5 and 1 microM, genistein and quercetin counteracted oestrogen- and transforming growth factor-alpha-promoted cell growth stimulation. Furthermore, genistein promoted transcription of the oestrogen-regulated genes pS2 and cathepsin-D, whereas quercetin interfered with the oestrogen-induced expression of the proteins. In in vitro binding experiments, genistein competed with oestradiol for binding to the oestrogen receptor (ER), but quercetin did not. Quercetin and genistein down-regulated cytoplasmic ER levels and promoted a tighter nuclear association of the ER, but only genistein was able to up-regulate progesterone receptor protein levels. In gel mobility assays, ER preincubation with oestradiol or with the two phyto-oestrogens led to the appearance of the same retarded band, excluding differences between the various complexes in binding to the consensus sequence. The data allowed us to conclude that quercetin acts like a pure anti-oestrogen, whereas genistein displays mixed agonist/antagonist properties, and to formulate a hypothesis on the possible mechanism of action of such phyto-oestrogens. PMID- 10376967 TI - Differentiation of human colon cancer cells changes the expression of beta tubulin isotypes and MAPs. AB - The human colon adenocarcinoma HT29-D4 cell line is an interesting model for studies on epithelial cell differentiation. Undifferentiated cells are malignant proliferating cells, whereas differentiated cells act like epithelial polarized cells. In the present study, we first characterized the action of taxoids on the microtubular network of HT29-D4 cells according to the state of differentiation. Microtubular bundles were found in undifferentiated cells but not in differentiated cells, even with 500-fold higher taxoid concentrations for 96 h. This finding led us to study changes in microtubules according to the polarity status of the cell. E-MAP-115 was expressed only in differentiated cells; expression of beta-tubulin isotypes was altered in them relative to undifferentiated cells. Classes I, II, III, IVa and IVb isotypes were expressed in both phenotypes; however, differentiated epithelial cells displayed a specific increase in class III beta-tubulin. Thus, the increase in expression of this beta tubulin isotype in differentiated cells is not restricted to neuronal cells. Moreover, these expression changes may reflect a higher stability of microtubular network in differentiated cells, which may explain the lower activity of anti microtubule agents, independently of the mitotic process. These results indicate that the composition of microtubules should be considered as one of the criteria involved in the response of tumour cells to chemotherapy with anti-microtubule agents. PMID- 10376968 TI - Overexpression of alpha(1,3)-fucosyltransferase VII is sufficient for the acquisition of lung colonization phenotype in human lung adenocarcinoma HAL-24Luc cells. AB - Metastatic human lung adenocarcinoma HAL-8Luc cells display an enhanced expression of alpha(1,3)-fucosyltransferases (alpha(1,3)-Fuc-Ts) compared with their non-metastatic counterpart HAL-24Luc cells. This correlates with an increased surface expression of Lewis(x) (Le(x))- and Lewis(a) (Le(a))-related molecules and an in vitro enhanced adhesive capacity to E-selectin-expressing endothelial cells (Martin-Satue et al (1998). Cancer Res 58: 1544-1550). In the present work we have stably transfected HAL-24Luc cells with the cDNAs for the alpha(1,3)-Fuc-TIV and VII enzymes and analysed by flow cytometry the expression of Le(x), sialyl-Le(x), sialyl-Le(x) dimeric, Le(a) and sialyl-Le(a). Fuc-TVII transfectants exclusively overexpress sialyl-Le(x) while Fuc-TIV-transfected cells only overexpress the Le(x) oligosaccharide. We show that solely Fuc-TVII transfectants are able to adhere to interleukin-1beta-stimulated HUVEC monolayers. We also demonstrate that Fuc-TVII overexpression in HAL-24Luc cells is sufficient for the acquisition of the lung colonization phenotype. This is the first report directly showing the contribution of an alpha(1,3)-Fuc-T to the metastatic behaviour of human lung adenocarcinoma cells. PMID- 10376970 TI - Frequent loss of expression or aberrant alternative splicing of P2XM, a p53 inducible gene, in soft-tissue tumours. AB - We investigated expression of a human p53-inducible gene, P2XM, a member of the P2X-receptor family of ATP-gated ion channels, in 56 human primary soft-tissue tumours including 47 sarcomas and nine benign tumors. Among the 47 sarcomas examined by reverse transcription polymerase chain reaction, 12 had lost expression of this gene and 22 revealed altered splicing patterns; among the nine benign tumours, four showed no expression of P2XM and three revealed aberrant splicing patterns involving transmembrane domains M1 and/or M2. As the aberrant transcripts lacked either or both of those domains, the protein products probably lacked normal function. We also looked for p53 mutations and mdm2 overexpression in the same panel of tumours and found them in 13 tumours, all but three of which had shown altered expression of P2XM. However, 31 (72%) of the 43 tumours that carried wild-type p53 without mdm2 overexpression had revealed aberrant P2XM expression. Our results suggest that disorder of P2XM expression may play a crucial role in the genesis of benign and malignant tumours in soft tissues, and that one or more genetic factors other than p53 or mdm2 contribute significantly to aberrant P2XM expression. PMID- 10376969 TI - G1 checkpoint protein and p53 abnormalities occur in most invasive transitional cell carcinomas of the urinary bladder. AB - The G1 cell cycle checkpoint regulates entry into S phase for normal cells. Components of the G1 checkpoint, including retinoblastoma (Rb) protein, cyclin D1 and p16INK4a, are commonly altered in human malignancies, abrogating cell cycle control. Using immunohistochemistry, we examined 79 invasive transitional cell carcinomas of the urinary bladder treated by cystectomy, for loss of Rb or p16INK4a protein and for cyclin D1 overexpression. As p53 is also involved in cell cycle control, its expression was studied as well. Rb protein loss occurred in 23/79 cases (29%); it was inversely correlated with loss of p16INK4a, which occurred in 15/79 cases (19%). One biphenotypic case, with Rb+p16- and Rb-p16+ areas, was identified as well. Cyclin D1 was overexpressed in 21/79 carcinomas (27%), all of which retained Rb protein. Fifty of 79 tumours (63%) showed aberrant accumulation of p53 protein; p53 staining did not correlate with Rb, p16INK4a, or cyclin D1 status. Overall, 70% of bladder carcinomas showed abnormalities in one or more of the intrinsic proteins of the G1 checkpoint (Rb, p16INK4a and cyclin D1). Only 15% of all bladder carcinomas (12/79) showed a normal phenotype for all four proteins. In a multivariate survival analysis, cyclin D1 overexpression was linked to less aggressive disease and relatively favourable outcome. In our series, Rb, p16INK4a and p53 status did not reach statistical significance as prognostic factors. In conclusion, G1 restriction point defects can be identified in the majority of bladder carcinomas. Our findings support the hypothesis that cyclin D1 and p16INK4a can cooperate to dysregulate the cell cycle, but that loss of Rb protein abolishes the G1 checkpoint completely, removing any selective advantage for cells that alter additional cell cycle proteins. PMID- 10376971 TI - BIRICODAR (VX-710; Incel): an effective chemosensitizer in neuroblastoma. AB - Clinical studies have suggested that both MDR1 and MRP may play a significant role in the chemosensitivity and outcome of neuroblastoma. To clarify the nature of multidrug resistance (MDR) in this tumour a series of six neuroblastoma cell lines have been characterized with regard to P-glycoprotein, MRP and LRP expression using immunocytochemistry and expression of MDR1, MRP, LRP and topoisomerase II genes using reverse transcription polymerase chain reaction (RT PCR). By RT-PCR, all lines expressed MRP, five expressed LRP and four expressed MDR1, but protein levels of each of these were variable. Chemosensitization to a range of MDR-associated drugs (vincristine, doxorubicin, etoposide, taxotere, topotecan) and non-MDR-associated drugs (cisplatin, melphalan) by three modulating agents, cyclosporin A, PSC 833 and the novel Biricodar (VX-710; Incel), was evaluated using a colourimetric cytotoxicity assay (MTS). Alteration of daunorubicin efflux by these agents was evaluated using FACS analysis. Clonogenic assay was used to study the influence of these chemosensitizers on vincristine cytotoxicity. Marked sensitization to vincristine was observed in MDR1-positive lines, and a similar but less consistent effect was seen with taxotere, doxorubicin and etoposide. With MRP-positive, MDR-negative lines, only VX-710 caused consistent sensitization. These data confirm MDR1 and MRP expression as contributory factors in chemoresistance in neuroblastoma and indicate that VX-710 may be a useful modulator of both mechanisms and worthy of clinical evaluation in this tumour. PMID- 10376972 TI - Apoptosis is induced in both drug-sensitive and multidrug-resistant hepatoma cells by somatostatin analogue TT-232. AB - Clinical resistance to chemotherapeutic drugs is an important problem in the treatment of cancer; the circumvention of resistance has become one of the basic goals of cancer therapy. The most frequent form of primary liver cancer is hepatocellular carcinoma, which is essentially refractory to chemotherapy. We earlier showed that TT-232, a new somatostatin analogue developed in our laboratory, exerted a strong antiproliferative effect both in vitro and in vivo, but no growth hormone release inhibitory or antisecretory activity. Here we report that TT-232 has a pronounced antiproliferative effect on differentiated and dedifferentiated, drug-sensitive and multidrug-resistant hepatocellular carcinoma cell lines. TT-232 induces apoptosis at comparable levels in all these hepatoma variants demonstrating that the multidrug resistance of hepatomas does not correlate with a reduced susceptibility to apoptosis induction. These results clearly reveal that the machinery involved in apoptosis is functional in both drug-sensitive and resistant hepatoma variants and can be activated by the somatostatin analogue TT-232. PMID- 10376973 TI - Increased doxorubicin uptake and toxicity in multicellular tumour spheroids treated with DC electrical fields. AB - Electrochemotherapy (ECT) is a new approach to the treatment of tumours. In the present study, multicellular prostate tumour spheroids were treated with non lethal direct current (DC) electrical fields, and uptake and toxicity of doxorubicin were investigated. An electrical field with a field strength of 500 Vm(-1) applied for a duration of 90 s resulted in neither reversible nor irreversible membrane breakdown as revealed by fluid phase uptake studies of the membrane impermeant tracer Lucifer yellow. However, treated spheroids showed an increased uptake of doxorubicin and, consequently, an increased toxicity following electrical field exposure. The electrical field raised intracellular reactive oxygen species (ROS) as revealed using 2',7'-dichlorofluorescein diacetate (H2DCFDA) as an indicator. ROS induced membrane lipid peroxidation since the lipid peroxidation end products malondialdehyde (MDA) and 4-hydroxy-2 (E)-nonenal (4-HNE) were detected after electrical field treatment. Moreover, lipid peroxidation decreased the lipid diffusion coefficient D from 4.2 x 10(-10) cm2 s(-1) to 2.7 x 10(-10) cm2 s(-1) in the control and treated sample, respectively, as revealed by fluorescence recovery after photobleaching (FRAP) experiments. The field effects could be mimicked by incubating spheroids with 100 nM hydrogen peroxide and were inhibited by the radical scavengers dehydroascorbate (DHA) and alpha-tocopherol (vitamin E), indicating that the increased uptake of doxorubicin after electrical field treatment is owing to lipid peroxidation and decreased membrane lipid mobility. Treatment of tumours with low intensity electrical fields may be useful to improve the cytotoxic capacity of anthracyclines. PMID- 10376974 TI - An anti-CD30 single-chain Fv selected by phage display and fused to Pseudomonas exotoxin A (Ki-4(scFv)-ETA') is a potent immunotoxin against a Hodgkin-derived cell line. AB - The human CD30 receptor is highly overexpressed on the surface of Hodgkin Reed Sternberg cells and has been shown to be an excellent target for selective immunotherapy using monoclonal antibody-based agents such as immunotoxins. To construct a new recombinant immunotoxin for possible clinical use in patients with Hodgkin's lymphoma, we have chosen the murine anti-CD30 hybridoma Ki-4 to generate a high-affinity Ki-4 single-chain variable fragment (scFv). Hybridoma V genes were polymerase chain reaction-amplified, assembled, cloned and expressed as a mini-library for display on filamentous phage. Functional Ki-4 scFv were obtained by selection of binding phage on the Hodgkin lymphoma-derived, CD30 expressing cell line L540Cy. The selected recombinant Ki-4 scFv was shown to specifically bind to an overlapping epitope on the CD30 antigen with binding kinetics similar to those of the original antibody. The Ki-4 scFv was subsequently fused to a deletion mutant of Pseudomonas exotoxin A (ETA'). The resulting immunotoxin Ki-4(scFv)-ETA' specifically binds to CD30+ L540Cy cells and inhibits the protein synthesis by 50% at a concentration (IC50) of 43 pM. This recombinant immunotoxin is a promising candidate for further clinical evaluation in patients with Hodgkin's lymphoma or other CD30+ malignancies. PMID- 10376975 TI - Enhanced cytotoxicity of mitomycin C in human tumour cells with inducers of DT diaphorase. AB - DT-diaphorase is a two-electron reducing enzyme that activates the bioreductive anti-tumour agent, mitomycin C (MMC). Cell lines having elevated levels of DT diaphorase are generally more sensitive to MMC. We have shown that DT-diaphorase can be induced in human tumour cells by a number of compounds, including 1,2 dithiole-3-thione. In this study, we investigated whether induction of DT diaphorase could enhance the cytotoxic activity of MMC in six human tumour cell lines representing four tumour types. DT-diaphorase was induced by many dietary inducers, including propyl gallate, dimethyl maleate, dimethyl fumarate and sulforaphane. The cytotoxicity of MMC was significantly increased in four tumour lines with the increase ranging from 1.4- to threefold. In contrast, MMC activity was not increased in SK-MEL-28 human melanoma cells and AGS human gastric cancer cells, cell lines that have high base levels of DT-diaphorase activity. Toxicity to normal human marrow cells was increased by 50% when MMC was combined with 1,2 dithiole-3-thione, but this increase was small in comparison with the threefold increase in cytotoxicity to tumour cells. This study demonstrates that induction of DT-diaphorase can increase the cytotoxic activity of MMC in human tumour cell lines, and suggests that it may be possible to use non-toxic inducers of DT diaphorase to enhance the efficacy of bioreductive anti-tumour agents. PMID- 10376976 TI - Effect of a cachectic factor on carbohydrate metabolism and attenuation by eicosapentaenoic acid. AB - The effect of a proteolysis-inducing factor (PIF), produced by cachexia-inducing tumours on glucose utilization by different tissues and the effect of pretreatment with the polyunsaturated fatty acid eicosapentaenoic acid (EPA), has been determined using the 2-deoxyglucose tracer technique. Mice receiving PIF showed a profound depression of body weight (2.3 g) over a 24-h period, which was completely abolished by pretreatment with a monoclonal antibody to PIF or by 3 days pretreatment with EPA at 500 mg kg(-1). Animals receiving PIF exhibited a marked hypoglycaemia, which was effectively reversed by both antibody and EPA pretreatment. There was an increase in glucose utilization by brain, heart and brown fat, but a decrease by kidney, white fat, diaphragm and gastrocnemius muscle after administration of PIF. Changes in organ glucose consumption were attenuated by either monoclonal antibody, EPA, or both. There was a decrease in 2 deoxyglucose uptake by C2C12 myoblasts in vitro, which was attenuated by EPA. This suggests a direct effect of PIF on glucose uptake by skeletal muscle. These results suggest that in addition to a direct catabolic effect on skeletal muscle PIF has a profound effect on glucose utilization during cachexia. PMID- 10376977 TI - Cytotoxic response of ovarian cancer cell lines to IFN-gamma is associated with sustained induction of IRF-1 and p21 mRNA. AB - Interferon-gamma (IFN-gamma) has some anti-tumour activity in human ovarian cancer. This cytokine inhibited proliferation in three of four ovarian cancer cell lines in vitro. We then compared the action of IFN-gamma in two cell lines, one sensitive and one resistant to its growth inhibitory effects. IFN-gamma signalling appeared normal in both cell lines, with stat1 DNA binding activity detectable at 30 min. Continuous exposure to IFN-gamma for 2-3 days was necessary for an irreversible effect on cell growth and apoptosis in cells sensitive to growth inhibition. During this time there was an increase in mRNA for the CKI p21, but no alterations in mRNA levels for other members of the CKI family. Maintenance of p21 mRNA required continuous mRNA synthesis. mRNA for the transcription factor IRF-1 was also induced in growth inhibited cells with similar kinetics to those observed for p21. Maximal induction of both p21 and IRF 1 mRNA was observed after 2-3 days IFN-gamma exposure as the cells became committed to cell death. There was also a rapid increase in p21 and IRF-1 mRNA in cells resistant to the growth inhibitory effects of IFN-gamma, but this increase was not maintained. Thus, continuous interaction with the IFN-gamma receptor, together with a sustained induction of p21 and IRF-1, is associated with growth inhibitory and apoptotic effects of IFN-gamma in ovarian cancer cells. PMID- 10376979 TI - Anti-tumour activity in vitro and in vivo of selective differentiating agents containing hydroxamate. AB - A series of hydroxamates, which are not metalloprotease inhibitors, have been found to be selectively toxic to a range of transformed and human tumour cells without killing normal cells (fibroblasts, melanocytes) at the same concentrations. Within 24 h of treatment, drug action is characterized by morphological reversion of tumour cells to a more normal phenotype (dendritic morphology), and rapid and reversible acetylation of histone H4 in both tumour and normal cells. Two hydroxamates inhibited growth of xenografts of human melanoma cells in nude mice; resistance did not develop in vivo or in vitro. A third hydroxamate, trichostatin A, was active in vitro but became inactivated and had no anti-tumour activity in vivo. Development of dendritic morphology was found to be dependent upon phosphatase activity, RNA and protein synthesis. Proliferating hybrid clones of sensitive and resistant cells remained sensitive to ABHA, indicating a dominant-negative mechanism of sensitivity. Histone H4 hyperacetylation suggests that these agents act at the chromatin level. This work may lead to new drugs that are potent, and selective anti-tumour agents with low toxicity to normal cells. PMID- 10376978 TI - Cisplatin anti-tumour potentiation by tirapazamine results from a hypoxia dependent cellular sensitization to cisplatin. AB - Tirapazamine (TPZ) is a new anticancer drug that is activated specifically at the low oxygen level typically found in solid tumours. It exhibits preferential cytotoxicity towards hypoxic cells and has been shown in preclinical studies with transplanted tumours and in phase II and III clinical trials to potentiate the anti-tumour efficacy of cisplatin without increasing its systemic toxicity. At present, the mechanism for this potentiation is unknown. Here we show that there is a schedule-dependent enhancement of cisplatin cytotoxicity by TPZ for cells in vitro that is similar to that seen with transplanted murine tumours. This cisplatin potentiation depends on the TPZ exposure being at oxygen concentrations below 1%, which are typical of many cells in tumours but not in normal tissues. Also, the interaction between TPZ and cisplatin does not occur in cells mutant in ERCC4, a protein essential for repair of DNA interstrand cross-links. Incubation of the cells with TPZ under hypoxia prior to cisplatin treatment increases cisplatin-induced DNA interstrand cross-links with kinetics suggesting that TPZ inhibits or delays repair of the DNA cross-links. In conclusion, we show that the tumour-specific potentiation of cisplatin cytotoxicity is likely the result of an interaction between TPZ and cisplatin at the cellular level that requires the low oxygen levels typical of those in solid tumours. The mechanism of the interaction appears to be through a potentiation of cisplatin-induced DNA interstrand cross links, possibly as a result of a diminished or delayed repair of these lesions PMID- 10376980 TI - Comparison of survival between malignant neuroendocrine tumours of midgut and pancreatic origin. AB - The survival of 64 consecutive patients with disseminated midgut carcinoid tumours was compared in a retrospective study with that of 25 consecutive patients with sporadic malignant endocrine pancreatic tumours treated according to similar surgical principles. The presence of hepatic metastases implied a worse prognosis in neuroendocrine tumours of pancreatic rather than midgut origin. This infers that these tumour types must be separated when treatments are evaluated. PMID- 10376981 TI - Aberrant DNA methylation of the p16INK4a gene in plasma DNA of breast cancer patients. AB - Hypermethylation of exon 1 of p16INK4a was examined in tumour and plasma DNA of a series of breast cancer patients. De novo methylation was observed in the tumours of eight patients (23%), and in plasma DNA in five (14%) of these eight patients. Our data show that de novo methylation of exon 1 of p16INK4a can be demonstrated in plasma DNA of breast cancer patients, a fact that provides additional evidence of the tumour-related origin of free plasma DNA in cancer patients. PMID- 10376982 TI - Measurement of urinary collagen cross-links indicate response to therapy in patients with breast cancer and bone metastases. AB - Objective assessment of response in bone metastases from breast cancer using radiological techniques takes up to 6 months of treatment to be certain of a response, and sclerotic metastases are not evaluable. Standard serum and urinary tumour markers may not always be utilized to predict response, as they may not be elevated, and therefore may not change on treatment. The development of the urinary pyridinoline cross-link assays which measure mature bone breakdown products have been shown to be highly sensitive and specific as a measure of bone change in osteoporosis. We have measured pyridinoline (Pyr) and deoxypyridinoline (Dpyr) cross-links sequentially in 36 breast cancer patients with bone metastases, to determine if the measurement of these analytes predicts response at an earlier stage than radiological assessment. Response was assessed by UICC criteria. Seventeen women responded to hormonal therapy, whilst 19 developed progressive disease. Both Pyr and Dpyr increased sequentially in women with progressive disease with changes becoming apparent by 8 weeks (P<0.03). In responding women, cross-link levels did not change significantly. Pyr and Dpyr were more sensitive and specific than the standard serum tumour marker CA 15-3. Urinary cross-link measurements provide a novel objective method of assessing response to treatment in women with bone metastases. Initial elevated urinary cross-link markers identify patients who tend not to respond to changes in hormonal therapy. PMID- 10376983 TI - Expression of oestrogen and progesterone receptors in gastric cancer: a flow cytometric study. AB - Increased expression of oestrogen (ER) and progesterone (PR) receptors have been reported in gastric adenocarcinoma, although results have been variable. Immunohistochemical staining methodologies, in particular in the detection of ER, have been inconsistent with many tumours being classified ER-negative. In this study we have used flow cytometry to quantify expression of ER and PR in gastric adenocarcinoma and examine their relationships with established prognostic indicators. Cytokeratin-positive cells obtained from tumour biopsies of 50 patients with gastric cancer and ten control patients were labelled with biotinylated ER or PR antibodies followed by streptavidin PE. Flow cytometry was seen to increase the detection of ER levels in gastric cancer with more receptor positive patients in this study than in results published to date. We believe this is related to the sensitivity of the flow cytometric assay with the detection of small shifts in ER level detected using cytokeratin gating. On analysis, the data showed no significant correlations with tumour stage and grade, and no differences were seen between normal mucosa and gastric cancer samples. PMID- 10376984 TI - Systematic review of cancer treatment programmes in remote and rural areas. AB - In an attempt to ensure high quality cancer treatment for all patients in the UK, care is being centralized in specialist centres and units. For patients in outlying areas, however, access problems may adversely affect treatment. In an attempt to assess alternative methods of delivering cancer care, this paper reviews published evidence about programmes that have set out to provide oncology services in remote and rural areas in order to identify evidence of effectiveness and problems. Keyword and textword searches of on-line databases (MEDLINE, EMBASE, HEALTHSTAR and CINAHL) from 1978 to 1997 and manual searches of references were conducted. Fifteen papers reported evaluations of oncology outreach programmes, tele-oncology programmes and rural hospital initiatives. All studies were small and only two were controlled, so evidence was suggestive rather than conclusive. There were some indications that shared outreach care was safe and could make specialist care more accessible to outlying patients. Tele oncology, by which some consultations are conducted using televideo, may be an acceptable adjunct. Larger and more methodologically robust studies are justified and should be conducted. PMID- 10376985 TI - Prognostic factors of oesophageal squamous cell carcinoma from the perspective of molecular biology. AB - Recent developments in molecular biology have revealed that several oncogenes, suppressor genes and adhesion molecules are involved in the development of oesophageal cancer; however, the role of these genes is still unknown. To evaluate which molecular biological factors are related to patients' prognosis and recurrence, we checked p53, p16, p21/Waf1, cyclin D1, Ki-67, epidermal growth factor receptor (EGFR), vascular endothelial growth factor (VEGF), Mdm2, Bcl2, E cadherin and MRP1/CD9 by means of immunohistochemical analysis in 116 cases of oesophageal cancer (R0). We also checked the regrowth capability of the primary cultures of the resected tumours and the effect of post-operative treatment. Although univariate analysis revealed that pN (pTNM), pT (pTNM), sex, cyclin D1, Ki-67, VEGF, E-cadherin and cell regrowth capability were prognostic factors, multivariate analysis revealed that pN (risk ratio (RR) 3.17), sex (RR 8.13), cell regrowth capability (RR 3.03) and E-cadherin (RR 0.30) were prognostic factors. Interestingly, step-wise analysis revealed that the following five factors were prognostic factors: pN (RR 5.74), sex (RR 3.14), cyclin D1 (RR 2.29), E-cadherin (RR 0.26) and cell regrowth capability (RR 1.94). Logistic regression analysis revealed that the risk factors of haematogenous recurrence were pN (odds ratio (OR) 8.97), cyclin D1 (OR 4.52) and EGFR (OR 0.18). On the other hand, the risk factor of lymph node recurrence was pN (OR 5.16). With regard to the effect of postoperative treatment, post-operative radiotherapy was a favourable risk factor (RR 0.43) and reduced the haematogenous recurrence (OR 0.18). Our data indicate that combination analysis using pN, sex, cyclin D1, E cadherin, EGFR and cell regrowth capability may be useful for the prediction of patient survival and recurrence. PMID- 10376987 TI - Clinico-pathological and treatment-related factors influencing survival in parotid cancer. AB - One hundred and three patients with primary parotid cancer treated surgically at the Christie Hospital, Manchester (1952-1992), were analysed to assess the influence on survival of prognostic and treatment-related factors. Thirty-seven patients were treated by surgery alone (SG), 66 received post-operative radiation (SG+RT). Median follow-up was 12 years, minimum 5 years. The 10-year disease specific survival rates for stage I, II and III/IV were 96%, 61% and 17% respectively (P < 0.0001). The various histological types segregated into three survival patterns: low-, intermediate-and high-grade with 10-year survival rates of 93%, 41% and 50% respectively (P < 0.0001). On multivariate analysis, the factors influencing risk of cancer death in order of importance were: tumour size > 4 cm (P < 0.001), presence of nodes (P = 0.001), histology of adenoid cystic carcinoma (P = 0.01), high-tumour grade (P = 0.02) and perineural involvement (P = 0.01). Neither the extent of surgery nor the operator influenced outcome. Overall, adjuvant RT significantly reduced locoregional recurrence (SG+RT 15% vs SG 43%; P = 0.002) but not survival, although on subanalysis, there was a trend to improved survival with large cancers and high-grade tumours. Long-term survival is determined primarily by tumour characteristics, namely clinical stage and grade. Post-operative RT contributes significantly to locoregional control and probably confers some survival advantage in high-risk patients. PMID- 10376988 TI - Incidence of childhood CNS tumours. PMID- 10376986 TI - Cyclin D1 expression in non-small-cell lung cancers: its association with altered p53 expression, cell proliferation and clinical outcome. AB - Cyclin D1, like p16INK4 (p16) and retinoblastoma (RB) proteins, participates in the cell cycle control at the G1-S transition. We have previously demonstrated altered p16 and RB protein status in non-small-cell lung cancers (NSCLCs) and their potential synergistic effect with altered p53 protein on proliferative activity (Kinoshita et al (1996) Cancer Res 56: 5557-5562). In the present study, cyclin D1 expression was studied by immunohistochemistry in the same cohort of 111 resected NSCLCs as in our previous study, and the amount of the cyclin D1 gene was analysed by Southern blot analysis in 29 NSCLCs. Cyclin D1 expression was analysed in relation to the status of p53, p16 and RB proteins, and proliferative activity determined by the Ki-67 index. It was also analysed in relation to survival of 77 patients with NSCLCs which were potentially curatively resected between 1990 and 1995. We found that: (1) cyclin D1 was expressed in 13 (11.7%) of 111 NSCLCs; (2) the cyclin D1 gene was neither significantly amplified nor rearranged; (3) cyclin D1 expression significantly correlated with altered p53 protein expression (P = 0.04), whereas it did not correlate with p16 and RB protein status; (4) proliferative activity tended to be higher in cyclin D1 positive (+) tumours than in cyclin D1-negative (-) tumours, although this difference was not statistically significant (P = 0.08); and (5) patients with cyclin D1+ tumours survived longer than patients with cyclin D1- tumours (5-year survival rates, 89% and 64% respectively, by the Kaplan-Meier method; P = 0.045 by the log-rank test), and cyclin D1 expression tended to be a favourable prognostic factor (P = 0.08 in univariate analysis). These findings suggest the involvement of cyclin D1 in the development and progression of NSCLCs, their proliferative activity and clinical outcome of NSCLC patients. PMID- 10376989 TI - Knockout of the glutamate dehydrogenase gene in bloodstream Trypanosoma brucei in culture has no effect on editing of mitochondrial mRNAs. AB - Glutamate dehydrogenase (GDH) was shown previously to bind the 3' oligo[U] tail of the mitochondrial guide RNAs (gRNAs) of Leishmania tarentolae, apparently in the dinucleotide pocket (Bringaud F, Stripecke R, Frech GC, Freedland S, Turck C, Byrne EM, Simpson L. Mol. Cell. Biol. 1997; 17:3915-3923). Bloodstream Trypanosoma brucei cells in culture represent a good system to investigate the genetic effects of knocking out kinetoplastid nuclear genes to test a role in RNA editing, since editing of several mitochondrial genes occurs but is dispensable for viability (Corell RA, Myler P, Stuart K. Mol. Biochem. Parasitol. 1994; 64:65 74 and Stuart K. In: Benne R, editor. RNA editing--the alteration of protein coding sequences of RNA. New York: Ellis Horwood, 1993:25-52). Both GDH alleles of bloodstream T. brucei in culture were replaced by drug resistant markers without any effect on viability. The ratios of edited to unedited mRNAs for several cryptogenes were assayed by primer extension analysis. The steady state abundances of these edited RNAs were unaffected by the double knockout. This evidence suggests that GDH may not play a role in the editing reaction in bloodstream trypanosomes in culture, but this conclusion is tentative since there could be redundant genes for any biological function. We employed a double allelic replacement technique to generate a tetracycline inducible conditional expression of an ectopic copy of the deleted gene in bloodstream trypanosomes in culture. We used this strategy for genes encoding mitochondrial proteins which are not required during this stage of the life cycle, but as a general strategy it should be appropriate for generation of conditional null mutants for essential genes as well. PMID- 10376990 TI - Theileria parva 104 kDa microneme--rhoptry protein is membrane-anchored by a non cleaved amino-terminal signal sequence for entry into the endoplasmic reticulum. AB - The 104 kDa microneme-rhoptry protein (p104) is the only known apical complex organelle-specific protein of Theileria parva. p104 exhibits striking structural similarities to circumsporozoite protein and sporozoite surface protein 2 of Plasmodium yoelii. Their primary sequences contain two hydrophobic segments, located at the amino-and the carboxy-terminus. The p104 amino-terminal hydrophobic region was suggested to be a signal peptide for entry into the endoplasmic reticulum and the extreme carboxy-terminal region to function as a membrane anchor. We have studied the biogenesis of p104 in a cell-free expression system and found that p104 is co-translationally transported into membranes derived from endoplasmic reticulum. The amino-terminal signal peptide is not cleaved off and anchors the protein in the membrane with the carboxy-terminal portion translocated into the lumen. We suggest that in vivo p104 is co translationally integrated into the membrane of the endoplasmic reticulum, from where it is further transported to the microneme-rhoptry complex. Thus, p104 appears to be a suitable marker to study the development of micronemes and rhoptries in T. parva. PMID- 10376991 TI - Characterization of the Ras homologue of Schistosoma mansoni. AB - Ras is a member of a super-family of guanine-binding or G-proteins. Ras functions as a molecular switch in the transduction of signals generated by the activation of a variety of cell surface receptors and relays the signals to downstream effectors. Little is known about signal transduction in schistosomes. In order for Schistosoma mansoni to survive different immune responses triggered by the host as well as to migrate from the site of penetration at the skin to the final destination in portal circulation, they must receive signals from the host environment and respond to them in a way that allows their survival. We have isolated the schistosome Ras cDNA by using sequence information of the schistosome Ras homologue submitted to the Genbank database. Analysis of the encoded peptide revealed 81% identity and 92% similarity with K-Ras from various species. Ras is a single copy gene as determined by quantitative hybridization experiments. The cDNA was cloned into pGEX-4T and the expressed peptide was used to generate specific antibody reagents. Affinity purified antibodies identified a 23 kDa native protein that localizes to the subtegument. Ras is not associated with the tegument. Ras is expressed in all the developmental stages of the parasite. However, Ras is over-expressed in female worms compared to males. Schistosome Ras was also shown to be post-translationally modified by addition of farnesyl isoprenoid moiety to the cysteine residue in the C-terminal box. Using a schistosome extract in vitro SmRas farnesylation was inhibited by the farnesyl transferase inhibitor, FTI-277, at concentrations comparable to those required to inhibit K-Ras processing. These initial studies on signal transduction in schistosomes should provide a solid basis for improving our understanding of schistosome-host interactions. PMID- 10376992 TI - Histidine-phosphorylation of succinyl CoA synthetase from Trypanosoma brucei. AB - The insect form of Trypanosoma brucei depends on respiration for its energy requirements. It contains a fully functional mitochondrion with a complete citric acid cycle. Most of its enyzmes have been characterized to date. The current study presents the characterization of the histidine phosphorylation activity of one of the few remaining enzymes, succinyl CoA synthetase. The trypanosomal enyzme was identified by partial purification, followed by direct protein sequencing. It is rapidly phosphorylated, presumably through auto phosphorylation, using either ATP or GTP as phosphate donors. The phosphorylation occurs exclusively on histidine residues. The histidine-bound phosphate can be donated to suitable phosphate acceptors in a rapid reaction. This phosphotransfer reaction is highly nucleotide selective, as only ADP, but none of the other nucleoside-diphosphates tested, can be used as a phosphate acceptor. PMID- 10376993 TI - Inhibition of succinyl CoA synthetase histidine-phosphorylation in Trypanosoma brucei by an inhibitor of bacterial two-component systems. AB - Recent drug screenings for new antibacterial drugs directed against histidine phospho-relay signalling pathways in bacteria have resulted in compounds which potently inhibit the histidine kinase activity of bacterial two-component systems. The present study demonstrates that one of these compounds, LY266500, is also a potent inhibitor of histidine phosphorylation in the unicellular eukaryotic parasite Trypanosoma brucei, both in vitro and in whole cells. In vitro, it inhibits histidine phosphorylation of mitochondrial succinyl CoA synthetase. LY26650 does not interfere with the phosphotransfer from the histidine-phosphorylated protein to ADP. In standardized cell culture tests, LY266500 potently inhibits the proliferation of the human pathogens T. brucei rhodesiense and Leishmania donovani. Since the inhibitory activity in vivo is life-cycle stage specific and correlates well with the mitochondrial activity in the different stages, the effect of LY266500 is most likely due to its specific inhibition of the mitochondrial succinyl CoA synthetase. PMID- 10376994 TI - Hemozoin stability and dormant induction of heme oxygenase in hemozoin-fed human monocytes. AB - Human monocytes avidly ingest malarial pigment, hemozoin. Phagocytosed hemozoin persists in the monocyte for a long time and modifies important monocyte functions. Stability of phagocytosed hemozoin may depend on modifications of the hemozoin heme moiety or reduced ability to express heme-inducible heme oxygenase. We show here that the spectral characteristics of alkali-solubilized hemozoin were identical to those of authentic heme, although hemozoin was solubilized by alkali much more slowly than authentic heme. Alkali-solubilized hemozoin was a substrate of microsomal rat heme oxygenase and bilirubin reductase, with bilirubin as the main final product. Hemozoin feeding to human monocytes did not induce heme oxygenase, but authentic heme and alkali-solubilized hemozoin supplemented to hemozoin-fed monocytes induced heme oxygenase and were degraded normally. Lysosomes isolated from hemozoin-fed monocytes released only traces of heme while lysosomes from erythrocyte-fed monocytes liberated considerable quantities of heme. Lack of heme release from hemozoin did not depend on proteolysis-resistant, heme-binding proteins, since lysosomal proteases fully degraded hemozoin-associated proteins but did not solubilize hemozoin. In conclusion, our data indicate that lack of induction of HO1 is due to the intrinsic structural characteristics of hemozoin and not to hemozoin-mediated impairment of the mechanism of HO1 induction. PMID- 10376995 TI - Energy-dependent efflux from Leishmania promastigotes of substrates of the mammalian multidrug resistance pumps. AB - We demonstrated the existence of three transport activities in promastigotes of Leishmania braziliensis, Leishmania guyanensis, and Leishmania mexicana. The first activity, an energy-dependent efflux of pirarubicin, was observed in all Leishmania species and inhibited by verapamil, by 2-[4-(diphenylmethyl)-1 piperazinyl]ethyl-5-(trans-4,6-dimethyl-1, 3,2-dioxaphosphorinan-2-yl)-2,6 dimethyl-4-(3-nitrophenyl)-3-py ridinecarboxylate P oxide (PAK104P) and by the phenothiazine derivatives: thioridazine, prochlorperazine, trifluoperazine, chlorpromazine and trifluoropromazine. The second activity, an energy-dependent efflux of calcein acetoxymethylester, was observed in all Leishmania species and inhibited by PAK104P and the same phenothiazine derivatives, but not by verapamil. The third activity, an energy-dependent efflux of calcein, was clearly detected in L. braziliensis and guyanensis and inhibited only by prochlorperazine and trifluoperazine. The fact that prochlorperazine and trifluoperazine inhibited the energy-dependent efflux of the three substrates suggests that these activities are mediated by the same transport system. It is noteworthy that the transport system identified in this study shares several properties with the mammalian multidrug resistance pump, MRP1. Pirarubicin, calcein acetoxymethylester and calcein are well known substrates of the MRP. Furthermore, the three types of inhibitors are also inhibitors of the MRP function. PMID- 10376996 TI - Characterization of a stage-specific Mr16000 schistosomular surface glycoprotein antigen of Schistosoma mansoni. AB - A 16 kDa Schistosoma mansoni schistosomular surface antigen (Sm16) was originally described as the target of a passively protective mAb (B3A). It appeared on the schistosomular surface after transformation of cercariae and was uniquely recognised by sera from animals exposed to attenuated cercariae. In this work sequential extractions of schistosomula with Triton X-114 and sodium dodecyl sulphate showed Sm16 to be an integral membrane structure which did not appear to be glycosylphosphatidylinositol-anchored as judged by experiments using phosphatidyl inositol-specific phospholipase C. The antigen was strongly reactive in Western blotting with rabbit irradiated vaccine sera. Sm16 was demonstrated in the hepatopancreas of S. mansoni-infected snails and was equally abundant in cercariae and mechanically- transformed schistosomula but was undetected in liver stage worms or eggs. Immunoelectron microscopy showed Sm16 to be localised, in cercariae, to what are believed to be subtegumental cell bodies packed with membraneous vesicles. Treatment with proteases and with sodium metaperiodate showed Sm16 to be a glycoprotein of which the epitope recognised by B3A was periodate sensitive. Two-dimensional electrophoresis gave a PI of 6. Neither the size or the recognition by B3A was affected by treatment with N-glycosidase F, endoglycosidase F or endo-alpha-N-acetylgalactosaminidase. Western blotting using a wide range of biotinylated lectins showed recognition only by peanut agglutinin and Ricinus communis agglutinin II (ricin). It is concluded that Sm16 has antigenic surface-exposed O-linked complex oligosaccharides which lack mannose/glucose, GlcNAc, L-fucose and sialic acid but contain terminal Gal beta (1-3) GalNAc and/or galactose. PMID- 10376997 TI - Effect of acidic pH on heat shock gene expression in Leishmania. AB - Temperature and pH shifts trigger differential gene expression and stage transformation in Leishmania. The parasites encounter dramatic fluctuations in the extra-cellular pH between the mid-gut of the sand fly (pH>8) and the phagolysosomal vacuole of mammalian macrophages (pH<6). The authors examined the effect of pH shifts on heat shock gene expression in Leishmania amazonensis and Leishmania donovani promastigotes. Acidic pH resulted in preferential stability of the hsp83 transcripts at 26 degrees C, but hsp transcripts were not preferentially translated as observed during heat shock. Pre-conditioning of promastigotes to acidic pH did not alter the temperature threshold for hsp synthesis but lead to an increase in hsp synthesis mainly in L. donovani at 37 degrees C, and to a slight decrease in the arrest of tubulin synthesis in L. amazonensis. The stage specific morphological alterations that take place in vitro correlated with the arrest in tubulin synthesis and occurred at different temperatures in L. donovani and L. amazonensis. PMID- 10376998 TI - Identification of the pro-mature processing site of Toxoplasma ROP1 by mass spectrometry. AB - The rhoptries are specialized secretory organelles that function during host cell invasion in the obligate intracellular parasite Toxoplasma gondii. All T. gondii rhoptry proteins studied to date are synthesized as pro-proteins that are then processed to their mature forms. To understand the role of the pro region in rhoptry protein function, we have precisely defined the processing site of the pro-region of the rhoptry protein ROP1. Efforts to determine such processing sites have been prevented by blocked N-termini of mature proteins isolated from T. gondii. To overcome this problem, we have used an engineered form of ROP1 and mass spectrometry to demonstrate that proROP1 is processed to its mature form between the glutamic acid at position 83 and alanine at position 84. These data also show that mature ROP1 lacks substantial post-translational modifications, a result which has important implications for targeting of rhoptry proteins. PMID- 10376999 TI - Identification of heparin as a ligand for the A-domain of Plasmodium falciparum thrombospondin-related adhesion protein. AB - Thrombospondin-related adhesion protein (TRAP) is a Plasmodium falciparum transmembrane protein that is expressed within the micronemes of sporozoites, and is implicated in host cell invasion and motility. Contained within the extracellular region of TRAP is an A-domain, a module found in a number of membrane, plasma and matrix proteins, that is often involved in ligand recognition. In order to determine the role of the TRAP A-domain, it has been expressed as a glutathione S-transferase fusion protein and its ligand binding compared with that of other characterised glutathione S-transferase A-domain fusion proteins. Using a solid phase assay to screen for binding to known A domain ligands, the TRAP A-domain was found to bind heparin. Binding to heparin appeared to be specific as it was saturable, and was inhibited by soluble heparin, fucoidan and dextran sulfate, but not by other negatively charged sulfated glycosaminoglycans such as chondroitin sulfates. Furthermore, unlike some A-domain ligand interactions, the A-domain of both TRAP and the leukocyte integrin, Mac-1, bound to heparin in the absence of divalent cations. It has been shown previously that another domain within TRAP, which is homologous to region II-plus of circumsporozoite protein, binds to sulfatide and to heparan sulfate on the immortalised hepatocyte line HepG2. The TRAP A-domain also bound to sulfatide and to HepG2 cells. Thus the A-domain shares certain binding properties already attributed to the region II-plus-like domain of TRAP, and may contribute to the binding of TRAP to heparan sulfate on hepatocytes. PMID- 10377000 TI - Selection and recovery of minor parasite populations expressing unique infected erythrocyte phenotypes. PMID- 10377001 TI - Isolation and subunit composition of the 20S proteasome of Giardia lamblia. PMID- 10377003 TI - Analysis of stage specificity of promoters in Plasmodium berghei using luciferase as a reporter. PMID- 10377002 TI - The Theileria annulata sporozoite and macroschizont polypeptide encoded by the spm1 gene shares phenylalanine-glycine motifs with nuclear pore proteins. PMID- 10377004 TI - Characterization of the genomic locus expressing the ERM-like protein of Echinococcus multilocularis. PMID- 10377005 TI - True hermaphroditism with partial duplication of chromosome 22 and without SRY. AB - We present the case of a patient with true hermaphroditism and partial duplication of chromosome 22. Cytogenetic evaluation showed no evidence of a Y chromosome in blood, skin, or gonadal tissue. Additional investigations using molecular probes showed no evidence of SRY. We conclude that there are genes on chromosome 22 that are involved in sex determination. PMID- 10377006 TI - Increased occurrence of cleft lip in glycogen storage disease type II (GSDII): exclusion of a contiguous gene syndrome in two patients by presence of intragenic mutations including a novel nonsense mutation Gln58Stop. AB - Genetic deficiency of lysosomal acid alpha-glucosidase (acid maltase) results in the autosomal recessive disorder glycogen storage disease type II (GSDII) in which intralysosomal accumulation of glycogen primarily affects function of skeletal and cardiac muscle. During an earlier review we noted 3 in 100 cases of GSDII with incidental description of cleft lip. In addition, we identified 2 of 35 GSDII patients referred to us for molecular studies with co-occurence of cleft lip, considerably greater than the estimated frequency of nonsyndromic cleft lip with or without cleft palate of 1 in 700 to 1,000. Because several lines of evidence support a minor cleft lip/palate (Cl/P) locus on chromosome 17q close to the locus for GSDII, we defined the molecular basis for the GSDII in these two patients to determine if they represented a contiguous gene syndrome. Patient I (of Dutch descent) was homozygous and the parents heterozygous for an intragenic deletion of exon 18 (deltaex18), common in Dutch patients. Patient II was heterozygous for delta525T, a mutation also common in Dutch patients and a novel nonsense mutation (172 [corrected] C-->T; Gln58Stop) in exon 2, the first coding exon. The mother was heterozygous for the delta525T and the father for the 172 [corrected] C-->T; Gln58Stop. The finding that both patients carried intragenic mutations eliminates a contiguous gene syndrome. Whereas the presence of cleft lip/cleft palate in a patient with GSDII could be coincidental, these co occurences could represent a modifying action of acid alpha-glucosidase deficiency on unlinked or linked genes that result in increased susceptibility for cleft lip. PMID- 10377007 TI - Bone mineral density and laboratory evaluation of a type II autosomal dominant osteopetrosis carrier. AB - Type II autosomal dominant osteopetrosis (ADO2) is an inherited disorder characterized by increased skeletal mass and characteristic abnormalities evident on radiography. Although previous investigators have described nonpenetrant individuals (carriers), it is not known whether carriers manifest subtle abnormalities. We hypothesized that ADO2 carriers would have an abnormality of osteoclast function that would lead to changes in bone mineral density (BMD), in serum tartrate-resistant acid phosphatase (TRAP), or in creatine kinase isoenzyme BB (CK-BB) levels that would permit carrier recognition. We identified a female carrier in a well-established ADO2 family and measured BMD, serum TRAP, and CK-BB concentrations. She had normal BMD, serum TRAP, and CK-BB concentrations. Thus, these measurements cannot be used to exclude carrier status in individuals who are seen for genetic counseling. However, measurements in other asymptotic carriers are necessary before concluding that these measurements are normal in all or most nonpenetrant individuals. PMID- 10377008 TI - Tibial agenesis, femoral duplication, and caudal midline anomalies. AB - Tibial agenesis with femoral duplication (Gollop-Wolfgang complex) and cloacal exstrophy are each rare malformations. Thus, their concurrence in an individual is an extremely rare event. We report on a patient born with distal duplication of the right femur, agenesis of the right tibia and hallux, cloacal exstrophy, and sacral defects. Review of the small group of cases reported with femoral duplication and tibial agenesis in association with caudal midline defects demonstrated a pattern of anomalies that while varying in presentation and severity was quite specific. We postulate that this disorder is related to misexpression of one or more distal HOX genes, potentially HOX10 or HOX11, leading to abnormal induction and proliferation of caudal mesenchyme. PMID- 10377009 TI - Phylogenetic analysis of the mitochondrial genome indicates significant differences between patients with Alzheimer disease and controls in a French Canadian founder population. AB - The activity of cytochrome oxidase (CO), the terminal enzyme of the mitochondrial electron transport chain, has been reported to be lower in the brains of Alzheimer disease (AD) patients. This suggests that a modification of mitochondrial DNA (mtDNA) may be responsible for this decrease of CO activity. Many mtDNA variants were found by different studies at a higher frequency in AD patients, suggesting that mtDNA variants could confer a genetic susceptibility to AD. In this study, we sequenced the entire mitochondrial genome region that encompasses the three CO genes and the 22 mitochondrial tRNA in 69 AD patients and 83 age-matched controls. We detected a total of 95 mtDNA variants. The allele frequencies of the majority of these variants were similar in patients and controls. However, a haplotype composed of three different modifications (positions: 5633, 7476, and 15812) was present in three of the 69 late-onset AD patients (4.3%) and also in 1 of 16 early-onset AD patients (6.2%) but not in control individuals. Given that one of these variants (15812) has already been shown to be associated with another neurodegenerative disease and that all three modifications are relatively conserved and their frequencies in the general population is only 0.1%, our data suggest that the presence of this haplotype may represent a risk factor for AD. We also found a significant association (P < 0.05) of two other variants at positions 709 (rRNA 12S) and 15928 (tRNA(Thr)). These two mtDNA variants are three times more frequent in control individuals compared with AD patients, suggesting that they may be protective against AD. PMID- 10377010 TI - Oesophageal atresia, related malformations, and medical problems: a family study. AB - Oesophageal atresia (OA) and tracheo-oesophageal fistula (TOF) are life threatening malformations of generally undefined cause. Previous reports of familial cases suggest a genetic contribution. The pattern of inheritance appears non-Mendelian, i.e., multifactorial. Individuals with OA/TOF often have other malformations and medical problems. The aim of this study was to determine the association in OA/TOF cases and healthy control subjects of associated malformations, midline defects, and medical conditions. We also investigate the relationships of these conditions in the relatives of the cases and controls. The results show that infants with OA/TOF frequently have VACTERL anomalies (vertebral, 17%; anal, 12%; cardiac, 20%; renal, 16%; limb, 10%) and other midline defects (cleft lip and palate, 2%; sacral dysgenesis, 2%; urogenital anomalies, 5%). The following medical problems were also reported: oesophageal dysmotility, 21%; gastro-oesophageal reflux, 22%; chest infections, 6%; and autonomic dysfunction, 0.5%. The first-degree relatives of children with OA are much more likely to have one of the aforementioned malformations or medical conditions when compared with the control group: one or more VACTERL anomalies (P < 0.01), gastro-oesophageal reflux (P < 0.05), recurrent respiratory infections (P < 0.05), and autonomic dysfunction (P < 0.001). The more distant relatives also show an increased incidence of such problems although in this case the data must be viewed with caution. The results confirm that the associated malformations and related medical problems occur significantly more frequently in the relatives of individuals with OA/TOF. These families may prove valuable for linkage analysis in an attempt to determine the genetics of OA/TOF. PMID- 10377011 TI - Early-infantile galactosialidosis: prenatal presentation and postnatal follow-up. AB - Galactosialidosis (GS) is an autosomal recessive condition caused by combined deficiency of the lysosomal enzymes beta-galactosidase and alpha-neuraminidase. The combined deficiency has been found to result from a defect in protective protein/cathepsin A (PPCA), an intralysosomal protein which protects these enzymes from premature proteolytic processing. The most severe form of GS, the early-infantile form, results in early onset of edema, ascites, visceromegaly, and skeletal dysplasia. We report a case of early-infantile GS in a male infant who presented with nonimmune fetal hydrops (NIH), "coarse" facial appearance, massive fluid-filled inguinal hernias, multiple telangiectasia, and diffuse hypopigmentation; he subsequently developed visceromegaly. The diagnosis of GS was confirmed biochemically and the defect in PPCA characterized at the protein level. Examination of fetal peripheral blood smears sampled at 30 weeks gestation demonstrated vacuolation of lymphocytes, suggesting blood film examination may be a useful screening tool for cases of NIH where a metabolic disorder is suspected. Skeletal radiography at birth demonstrated punctate epiphyses of the femora, calcanei, and sacrum. We present a discussion of and differential diagnosis for this radiographic finding. To the best of our knowledge, this is the first case of early-infantile GS presenting with stippled epiphyses. PMID- 10377012 TI - Sanjad-Sakati and autosomal recessive Kenny-Caffey syndromes are allelic: evidence for an ancestral founder mutation and locus refinement. AB - The Sanjad-Sakati syndrome (SSS; MIM241410), an autosomal recessive trait characterized by congenital hypoparathyroidism, growth and mental retardation, seizures, and a characteristic physiognomy, was recently linked to chromosome area 1q42-q43. SSS resembles the autosomal recessive form of Kenny-Caffey syndrome (KCS; MIM244460), with similar manifestations but lacking osteosclerosis. Since KCS was recently linked to the region 1q42-q43, the possibility that this disorder is allelic with SSS was considered. Eight Sanjad Sakati families from Saudi Arabia were genotyped with polymorphic short tandem repeat markers from the SSS/KCS critical region. A maximum multipoint LOD score of 14.32 was obtained at marker D1S2649, confirming linkage of SSS to the same region as autosomal recessive KCS. Haplotype analysis refined the critical region to 2.6 cM and identified a rare haplotype present in all the SSS disease alleles, indicative of a common founder. In addition to the assignment of the Saudi SSS and Kuwaiti KCS syndromes to overlapping genetic intervals, comparison of the haplotypes unexpectedly demonstrated that the diseases shared an identical haplotype. This finding, combined with the clinical similarity between the two syndromes, suggests that the two conditions are not only allelic but are also caused by the same ancestral mutation. PMID- 10377013 TI - Severe achondroplasia with developmental delay and acanthosis nigricans (SADDAN): phenotypic analysis of a new skeletal dysplasia caused by a Lys650Met mutation in fibroblast growth factor receptor 3. AB - We previously discovered a novel missense mutation (Lys650Met) in the tyrosine kinase domain of the fibroblast growth factor receptor 3 (FGFR3) gene in four unrelated individuals with a condition we called "severe achondroplasia with developmental delay and acanthosis nigricans" (SADDAN) [Tavormina et al., 1999: Am. J. Hum. Genet. 64:722-731]. Here we present a more detailed clinical account of the SADDAN phenotype. The FGFR3 Lys650Met mutation results in severe disturbances in endochondral bone growth that approach and overlap those observed in thanatophoric dysplasia, type I. However, this mutation is most often compatible with survival into adulthood. Other unusual bone deformities, such as femoral bowing with reverse (i.e., posterior apex) tibial and fibular bowing and "ram's horn" bowing of the clavicle, are also seen in some patients. In addition to skeletal dysplasia, progressive acanthosis nigricans, and central nervous system structural anomalies, seizures and severe developmental delays are observed in surviving SADDAN patients. Despite its location within the same FGFR3 codon as the thanatophoric dysplasia type II mutation (Lys650Glu) and a similar effect on constitutive activation of the FGFR3 tyrosine kinase, the Lys650Met is not associated with cloverleaf skull or craniosynostosis. PMID- 10377014 TI - Investigation of maternal blood enriched for fetal cells: role in screening and diagnosis of fetal trisomies. AB - Prenatal diagnosis of chromosomal abnormalities relies on assessment of risk followed by invasive testing in the group with highest risk. Assessment of risk by a combination of maternal age and fetal nuchal translucency and invasive testing in the 5% of the population with the highest risk would identify about 80% of trisomy 21 pregnancies. Preliminary reports suggest that chromosomal abnormalities can also be diagnosed by fluorescent in situ hybridization (FISH) in maternal blood enriched for fetal cells. This study examines the potential role of this method on the prenatal diagnosis of fetal trisomies. Maternal blood was obtained before invasive testing in 230 pregnancies at 10-14 weeks of gestation. After enrichment for fetal cells, by triple density centrifugation and anti-CD71 magnetic cell sorting, FISH was performed and the proportion of cells with positive signals in the chromosomally normal and abnormal groups was determined. Fetal karyotype was normal in 150 cases and abnormal in 80 cases, including 36 with trisomy 21. Using a 21 chromosome-specific probe, three-signal nuclei were present in at least 5% of the enriched cells from 61% of the trisomy 21 pregnancies and in none of the normal pregnancies. For a cut-off of 3% of three-signal nuclei the sensitivity for trisomy 21 was 97% for a false positive rate of 13%. Similar values were obtained in trisomies 18 and 13 using the appropriate chromosome-specific probe. Examination of fetal cells from maternal blood may provide a noninvasive prenatal diagnostic test for trisomy 21 with the potential of identifying about 60% of affected pregnancies. Alternatively, this technique can be combined with maternal age and fetal nuchal translucency as a method of selecting the high-risk group for invasive testing. Potentially, 80% of trisomy 21 pregnancies could be identified after invasive testing in less than 1% of the pregnant population. PMID- 10377015 TI - Mulibrey nanism and Wilms tumor. PMID- 10377016 TI - Infant with manifestations of oto-palato-digital syndrome type II and of Melnick Needles syndrome. PMID- 10377017 TI - Follow-up of an adult with Keutel syndrome. PMID- 10377018 TI - Maternal vitamin use, infant C677T mutation in MTHFR, and isolated cleft palate risk. PMID- 10377019 TI - Rare sex chromosome aneuploidies in humans: report of six patients with 48,XXYY, 49,XXXXY, and 48,XXXX karyotypes. PMID- 10377020 TI - Radiographic documentation does not permit the diagnosis of MNS. PMID- 10377021 TI - Genetic heterogeneity of isolated noncompaction of the left ventricular myocardium. PMID- 10377022 TI - Paradoxical emboli: the relationship between patent foramen ovale, deep vein thrombosis and ischaemic stroke. PMID- 10377023 TI - Realistic expectations for patients with stent-graft treatment of abdominal aortic aneurysms. Results of a European multicentre registry. AB - OBJECTIVE: the outcomes for patients after endovascular treatment of abdominal aortic aneurysm (AAA) are determined primarily by the endpoints of death and endoleaks, the latter representing continued risk of rupture. The data of a multicentre registry were analysed with regard to the early outcome of stent graft procedures for AAA and the complications associated with this treatment. In addition, the results during follow-up were analysed by determining mortality and endoleak development as separate endpoints and as a combined endpoint defined as endoleak-free survival. SETTING: 38 European institutions of Vascular Surgery collaborating in a multicentre registry project. PATIENTS AND METHODS: 899 patients with AAA underwent between May 1994 and March 1998 elective endovascular repair (818 men and 81 women; mean age 69 years). 80 (8.9%) of the patients had medical conditions that excluded them from open repair. 818 (91%) of patients had a bifurcated device, 63 (7%) had a straight tube graft, and only 18 (2%) had an aorto-uni-iliac device. Clinical examination and contrast-enhanced computed tomography was performed at fixed follow-up intervals to assess increase or decrease of the maximum transverse diameter (MTD). Endoleaks observed at follow up were discriminated into persistent endoleak and temporary endoleak. The latter is defined as single time observed endoleaks or with two or more negative imaging studies between observed endoleaks. Life-table analyses were used to calculate the rates of freedom-from-endoleak (no endoleak at any time), freedom-from persistent endoleak (no persistent endoleak), patient survival, and persistent endoleak-free-survival. RESULTS: the median follow-up of this patient series was 6.2 months. The ratio between observed and expected follow-up data was 82% for the overall follow-up period. However, at 18 months of follow-up this rate was only 45%. The number of patients followed during this period was sufficient to allow statistically meaningful assessment. The MTD in patients with temporary endoleaks demonstrated a significant decrease at 6 to 12 months compared to preoperative values (mean 57 and 53 respectively, p =0.004). In patients with persistent endoleaks there was no change between the preoperative and 6-month MTD (mean 57 and 60 mm respectively). At 6 and 18 months freedom-from-endoleak was 83% and 74% and freedom-from-persistent endoleak was 93% and 90%, respectively. The 18-month cumulative patient survival was 88% and the main outcome measure, the persistent endoleak-free-survival was 79%. CONCLUSIONS: the MTD decreases in patients with temporary endoleak, but not in patients with persistent endoleak. Therefore, the use of the rate of freedom-from-persistent endoleak, reflecting absence of persisting endoleaks to estimate the prognosis with regard to the AAA, is justified. Determining persistent endoleak-free survival appears a rational approach to provide a realistic outlook for patients with stent-grafted AAA. The observed 18-month endoleak-free survival reflects a satisfactory mid-term result. PMID- 10377024 TI - Interplay of maturation-promoting factor and mitogen-activated protein kinase inactivation during metaphase-to-interphase transition of activated bovine oocytes. AB - The objective of the present study was to examine the activity changes in histone H1 kinase (also known as maturation-promoting factor [MPF]) and mitogen-activated protein kinase (MAPK) and their constituent proteins in in vitro-matured bovine oocytes after in vitro fertilization (IVF) or after parthenogenetic activation induced by calcium ionophore A23187 alone or by the ionophore followed by either 6-dimethylaminopurine (6-DMAP) or cycloheximide (CHX). Inactivation of both H1 kinase and MAPK occurred after both A23187+6-DMAP treatment and IVF; inactivation of H1 kinase preceded inactivation of MAPK. However, MAPK was inactivated much earlier in 6-DMAP-treated oocytes. Further analysis of constituent cell cycle proteins of these kinases by Western blot showed that A23187 alone could not induce changes in cdc2, cdc25, or ERK2 but induced reduction of cyclin B1. IVF and A23187+CHX induced similar changes: cyclin B1 was destroyed shortly after activation followed by accumulation of cyclin B1, phosphorylation of cdc2, and dephosphorylation of ERK2 at pronuclear formation 15 h after activation. No change in cdc25 was observed at this time. In contrast, A23187+6-DMAP treatment resulted in earlier phosphorylation of cdc2 and dephosphorylation of ERK2 at 4 h after treatment when the pronucleus formed. Moreover, accumulation of both cdc25 and cyclin B1 was detected at 15 h. Microinjection of ERK2 antibody into A23187 treated oocytes resulted in pronuclear formation. In conclusion, activation of bovine oocytes with 6-DMAP led to earlier inactivation of MAPK, while CHX induced inactivation of MAPK parallel to that following sperm-induced oocyte activation. Destruction of cyclin B is responsible for inactivation of MPF, while phosphorylation of cdc2 is likely responsible for maintaining its low activity. Inactivation of MAPK is closely associated with pronuclear development regardless of the activation protocol used. PMID- 10377025 TI - Endogenous nitric oxide suppresses rat myometrial connexin 43 gap junction protein expression during pregnancy. AB - Nitric oxide (NO) synthase (NOS) is active in the gravid uterus, and its activity decreases prior to the onset of parturition. We tested the hypothesis that NO helps maintain uterine quiescence by suppressing the expression of genes necessary for parturition. Pregnant rats (18 days gestation) were treated with inducible NOS (iNOS) inhibitor N-iminoethyl-L-lysine (NIL) or endothelial NOS inhibitor nitro-L-arginine methyl ester (L-NAME); 24 h later, uteri were analyzed for myometrial connexin 43 (Cx43) protein by immunoblotting and mRNA by Northern analysis. Myometrial oxytocin receptors (OTR) were measured by radioligand binding, and decidual prostaglandin H synthase (PGHS) protein by immunoblotting. Uterine NOS blockade was verified by NOS activity assay. We found that NIL, but not L-NAME, significantly increased myometrial Cx43 protein to parturitional levels with treatment at 19 but not 17 days gestation. Steady state mRNA concentrations were not changed at 24 h. NOS inhibition did not increase the concentrations of OTR, or PGHS protein, nor did it decrease maternal serum progesterone. We conclude that endogenous uterine NO from iNOS suppresses myometrial Cx43 gap junction protein expression during rat pregnancy. Although the exact mechanism is unknown, an increase of uterine wall stretch due to inhibition of relaxation could account for increased Cx43 gene transcription. PMID- 10377026 TI - Expression of matrix metalloproteinases and their tissue inhibitor messenger ribonucleic acids in macaque periovulatory granulosa cells: time course and steroid regulation. AB - Progesterone appears essential for ovulation and luteinization of the primate follicle, but specific gene targets of progesterone action remain elusive. Limited evidence supports a role for progesterone in the induction of collagenolytic activity in the periovulatory follicle of primate and nonprimate species. This study was designed to elucidate the pattern of expression and progesterone regulation of mRNAs for the matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in macaque granulosa cells during controlled ovarian stimulation cycles before (0 h) and after (up to 36 h) administration of an ovulatory hCG bolus. Levels of mRNAs for interstitial collagenase, gelatinase A, matrilysin, TIMP-1 and TIMP-2 increased (p < 0.05) within 12 h of hCG, while gelatinase B mRNA increased later, by 36 h after hCG. Administration of a 3beta hydroxysteroid dehydrogenase inhibitor (Trilostane [TRL]) during hCG treatment decreased (p < 0.05) mRNA levels for interstitial collagenase, gelatinase B, matrilysin, TIMP-1, and TIMP-2. Progestin (R5020) replacement during hCG+TRL treatment returned interstitial collagenase and TIMP-1 mRNAs to control levels. These data suggest that one action of progesterone, and possibly other steroids, in the cascade of events leading to ovulation and luteinization of the primate follicle is to regulate the expression of specific ovarian proteases and protease inhibitors. PMID- 10377027 TI - Developmentally regulated loss and reappearance of immunoreactive somatic histone H1 on chromatin of bovine morula-stage nuclei following transplantation into oocytes. AB - One difference between chromatin of bovine oocytes and blastomeres is that somatic subtypes of histone H1 are undetectable in oocytes and are assembled onto embryonic chromatin during the fourth cell cycle. We investigated whether this chromatin modification is reversed when nuclei containing somatic H1 are transplanted into ooplasts. Donor nuclei obtained from morula-stage bovine embryos were fused to ooplasts at different times before and after parthenogenetic activation of the ooplasts. After fusion, immunoreactive H1 became undetectable, and the loss occurred more rapidly when fusion was performed near the time of ooplast activation compared with several hours after activation, when the host oocytes were at a stage corresponding to interphase. Although the loss of immunoreactive H1 occurred independently of DNA replication and transcription, exposure of reconstructed oocytes to cycloheximide or 6 dymethylaminopurine (6-DMAP) delayed the loss of immunoreactive H1 from transplanted nuclei. During further development of nuclear-transplant embryos, somatic H1 remained undetectable at the 2- and 4-cell stages, and it reappeared on the chromatin at the 8- to 16-cell stage, as previously observed in unmanipulated embryos. We conclude that factors in oocyte cytoplasm are able to modify morula chromatin so that somatic H1 becomes undetectable, and that the amount or activity of these factors declines over time in activated ooplasts. PMID- 10377028 TI - Follicle deviation and intrafollicular and systemic estradiol concentrations in mares. AB - By definition, follicle deviation begins on the day the two largest follicles of a wave begin to differ in growth rates. The relationships between follicle deviation and intrafollicular and systemic estradiol concentrations were studied in ponies, using a two-follicle model in which all but the two largest follicles were ablated. A 20-microliter sample of follicular fluid was obtained from each of the two follicles by transvaginal ultrasonography. In experiment 1, the two follicles were sampled when the larger follicle reached 15 mm. No differences (p > 0.05) in post-sampling follicle characteristics were found between control (n = 6) and sampled (n = 8) groups except that the growth rate was slower (p < 0.01) in the larger follicle between the day of sampling and the next day (0.7 +/- 0.7 mm per day) than in the controls (3.3 +/- 0.3 mm per day). The growth rates between 2 and 5 days after sampling were not different between groups. Follicular fluid estradiol-17beta concentrations were higher (p < 0.007) in the larger follicle (460 +/- 67 ng/ml; diameter, 16.4 +/- 0.4 mm) than in the smaller follicle (322 +/- 50 ng/ml; diameter, 14.6 +/- 0.6 mm). In experiment 2, the pair of follicles was sampled when the larger follicle reached 15 mm, 20 mm, or 25 mm (n = 5 per group). There were no significant differences among the three groups for day of deviation and diameters of larger and smaller follicles at deviation. The difference in diameter between the larger and smaller follicles was similar for the 15-mm (2.2 +/- 0.9 mm) and 20-mm (3.1 +/- 1.0 mm) groups, but the difference between follicles for the 25-mm group (7.9 +/- 1.2 mm) was greater (p < 0.004) than for the other two groups. In contrast, the differences in estradiol concentrations between the larger and smaller follicles increased (p < 0.0001) progressively for the 15-mm (13.0 +/- 86.8 ng/ml), 20-mm (722.0 +/- 173.8 ng/ml), and 25-mm (1873.5 +/- 310.3 ng/ml) groups. The first significant (p < 0.007) increase in systemic estradiol occurred between the day before and the day of the beginning of deviation. Detection of an increased difference in estradiol concentrations between the two follicles before the detection of a change in differences in diameter suggests, on a temporal basis, that estradiol is a candidate for involvement in the mechanism that leads to follicle-diameter deviation in mares. PMID- 10377029 TI - Prostaglandins and leukotriene B4 are potent inhibitors of 11beta-hydroxysteroid dehydrogenase type 2 activity in human choriocarcinoma JEG-3 cells. AB - The 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2) is responsible for the inactivation of glucocorticoids. This is the predominant isozyme in the human placenta, where it is proposed to protect the fetus from high levels of maternal cortisol. In the present study, we examined the effects of eicosanoids on the activity of 11beta-HSD2 in human choriocarcinoma JEG-3 cells, a well-established model for placental trophoblasts. Treatment of JEG-3 cells for 24 h with either prostaglandin (PG) E2 or F2alpha attenuated 11beta-HSD2 activity ( approximately 40%). Paradoxically, indomethacin, an inhibitor of cyclooxygenases, inhibited (approximately 40%) rather than stimulated the activity of this enzyme. This indicated that the arachidonic acid metabolism may be diverted to other pathway(s), the products of which may inhibit 11beta-HSD2 activity. To determine whether the lipoxygenase pathways were involved, the cells were treated with nordihydroguaretic acid (NDGA), a blocker of all three (5-, 12-, and 15-) lipoxygenases. NDGA caused a 3-fold increase in 11beta-HSD2 activity. To further delineate which specific lipoxygenase pathway was involved, the cells were incubated with zileuton, a selective inhibitor of 5-lipoxygenase. This resulted in a similar increase in 11beta-HSD2 activity, suggesting that the products of this pathway (e.g., leukotrienes) may be involved. Given that leukotriene B4 (LTB4) is the most biologically active product of the 5-lipoxygenase pathway, we treated the cells with LTB4, which inhibited 11beta-HSD2 activity in a time- and dose-dependent manner with a maximal effect (60% reduction) at 10 nM for 9 h. Semiquantitative reverse transcription-polymerase chain reaction analysis revealed that 11beta-HSD2 mRNA levels were not altered by the addition of LTB4, PGE2, or PGF2alpha, indicating an effect at the posttranscriptional level. In conclusion, these results demonstrate that prostaglandins and LTB4 are potent inhibitors of 11beta-HSD2 activity in JEG-3 cells, suggesting that placental 11beta-HSD2 activity is modulated by these locally produced eicosanoids. This is the first time that the products of arachidonic acid metabolism have been found to regulate the activity of 11beta-HSD2. PMID- 10377030 TI - Embryo survival, and fetal and placental growth following elevation of maternal estradiol blood concentrations in the rat. AB - High doses of estrogens cause embryonic mortality, and fetal and placental growth retardation in rats. This study addresses the physiological relevance of such findings. Estradiol benzoate (EB), by s.c. injection, or estradiol-17beta (E2), delivered by a miniosmotic pump, raised maternal E2 concentrations from only slightly above control values to 5-fold. EB (1 microgram/day) over Days 6-13, 8 13, and 11-13, and continuous infusion of E2 (15 ng/h; Days 10-13) reduced fetal survival to 0%, 0%, 22%, and 75%, respectively. Single injections of EB showed that its lethal effect declined rapidly over Days 9 (44% survival) to 13 (90% survival). Embryos died within 48 h, but death was not due to luteal failure since progesterone levels were maintained and progesterone administered with EB did not reduce mortality. Administration of EB at 1 microgram/day (Days 14-21) or E2 at 40 ng/h (Days 13-16) retarded fetal and placental growth but did not affect survival. The rat embryo is highly sensitive to elevated maternal estradiol concentrations over much of gestation. The early lethal effect implies that endogenous E2 production is carefully regulated to maintain pregnancy; the latter growth-retarding effect suggests that E2 may have a role in the normal control of fetal growth. PMID- 10377031 TI - Binding characteristics of estrogen receptor (ER) in Atlantic croaker (Micropogonias undulatus) testis: different affinity for estrogens and xenobiotics from that of hepatic ER. AB - An estrogen receptor (ER) was identified in cytosolic and nuclear fractions of the testis in a marine teleost, Atlantic croaker (Micropogonias undulatus). A single class of high affinity, low capacity, and displaceable binding sites was identified by saturation analysis, with a Kd of 0.40 nM in cytosolic extracts and a Kd of 0.33 nM in nuclear extracts. Competition studies demonstrated that the receptor was highly specific for estrogens (diethylstilbestrol > estradiol >> estriol = estrone) and also bound several antiestrogens. Testosterone and 5alpha dihydrotestosterone had much lower affinities for the receptor, whereas no displacement of specific binding occurred with 11-ketotestosterone or any of the C21 maturation-inducing steroids. A variety of xenoestrogens, including o,p' dichlorodiphenyltrichloroethane (DDT), chlordecone (Kepone), nonylphenol, hydroxylated polychlorinated biphenyls (PCBs), and the mycotoxin zearalenone, bound to the receptor with relatively low binding affinities, 10(-3) to 10(-5) that of estradiol. A comparison of the binding affinities of various ligands for the testicular ER and the hepatic ER in this species revealed that the testicular ER was saturated at a lower [3H]estradiol concentration (1 nM vs. 4 nM). The binding affinities of several compounds, including testosterone and nafoxidine, exhibited marked differences for the two ERs; and most of the estrogens and xenoestrogens tested had higher binding affinities for the testicular receptor. Minor amounts of estradiol (0.12 ng/g tissue/h) were produced by testicular tissue fragments incubated in vitro, and estradiol was detected in male Atlantic croaker plasma. The identification of a testicular ER and evidence that estradiol is produced by the testes in croaker suggest that estrogens participate in the hormonal control of testicular function in teleosts. PMID- 10377032 TI - Human sperm proteome: immunodominant sperm surface antigens identified with sera from infertile men and women. AB - The objective of this study was to identify those immunodominant sperm antigens recognized by antisperm antibodies (ASA) in the serum samples of infertile men and women. High-resolution two-dimensional gel electrophoresis was employed to separate human sperm proteins using isoelectric focusing or nonequilibrium pH gradient electrophoresis, followed by PAGE. Serum samples from 15 infertile male subjects and 6 infertile female subjects that contained ASA as assayed by the immunobead binding test (IBT) were analyzed by Western blotting followed by enhanced chemiluminescence (ECL). Serum samples from 10 fertile subjects (5 males and 5 females) that were ASA negative by IBT were used as controls. The ECL blots were analyzed by computer scanning to compare the immunoreactivity between serum samples from fertile and infertile subjects and to identify the antigens unique to the sera of the infertile subjects; 98 sperm auto- and iso-antigenic protein spots were recognized by sera from infertile males and females but not from fertile subjects. Based on vectorial labeling with 125I at the sperm surface, a subset of 6 auto- and iso-antigens was identified as possibly relevant to antibody-mediated infertility. PMID- 10377033 TI - Germ cell apoptosis in the testes of Sprague Dawley rats following testosterone withdrawal by ethane 1,2-dimethanesulfonate administration: relationship to Fas? AB - Germ cell apoptosis, which occurs normally during spermatogenesis, increases after testosterone withdrawal from the testis. The molecular mechanism by which this occurs remains uncertain. The Fas system has been implicated as a possible key regulator of apoptosis in various cells: binding of Fas ligand (FasL), a type II transmembrane protein, to Fas, a type I transmembrane receptor protein, triggers apoptosis in cells expressing Fas. Recently, Fas has been localized to germ cells, and FasL to Sertoli cells, within the rat testis. We hypothesized that Fas protein content would rise in response to reduced levels of testosterone as part of a suicide pathway that would result in germ cell apoptosis. To test this hypothesis, ethane 1,2-dimethanesulfonate (EDS), a Leydig cell toxicant, was used to kill Leydig cells and thus reduce intratesticular testosterone levels in Sprague Dawley rats. Apoptosis was examined in situ and biochemically, and Fas protein content in the testis was monitored by Western blot analysis. We show that EDS injection results in the following sequence of events: apoptotic death of Leydig cells by a mechanism that does not involve Fas; reduced testosterone; increased testicular Fas content; and germ cell apoptosis. These results suggest that Fas may play a role in the apoptotic death of germ cells that results from reduced intratesticular testosterone levels, and that testosterone may play a role in germ cell survival via its suppression of Fas. PMID- 10377034 TI - Roles of bicarbonate, cAMP, and protein tyrosine phosphorylation on capacitation and the spontaneous acrosome reaction of hamster sperm. AB - Capacitation is a prerequisite for successful fertilization by mammalian spermatozoa. This process is generally observed in vitro in defined NaHCO3 buffered media and has been shown to be associated with changes in cAMP metabolism and protein tyrosine phosphorylation. In this study, we observed that when NaHCO3 was replaced by 4-(2-hydroxyethyl)1-piperazine ethanesulfonic acid (HEPES), hamster sperm capacitation, measured as the ability of the sperm to undergo a spontaneous acrosome reaction, did not take place. Addition of 25 mM NaHCO3 to NaHCO3-free medium in which spermatozoa had been preincubated for 3.5 h, increased the percentage of spontaneous acrosome reactions from 0% to 80% in the following 4 h. Addition of anion transport blockers such as 4,4'-diiso thiocyano-2, 2'-stilbenedisulfonate (DIDS) or 4-acetomido-4' isothiocyanatostilbene-2,2'-disulfonic acid (SITS) to the NaHCO3-containing medium inhibited the acrosome reaction, with maximal inhibition at 600 microM, and with an EC50 of 100 microM. Increasing either extracellular or intracellular pH did not induce the acrosome reaction in NaHCO3-free medium. In contrast, addition of 500 microM dibutyryl cAMP (dbcAMP), alone or together with 100 microM 1-methyl-3-isobutylxanthine (IBMX), induced the acrosome reaction in spermatozoa incubated in NaHCO3-free medium. These compounds also partially reversed the inhibition of the acrosome reaction caused by the DIDS or SITS in complete medium. In contrast to these results, IBMX or dbcAMP did not induce acrosome reactions in cells incubated in Ca2+-free medium. When hamster sperm were incubated in the absence of NaHCO3 or in the presence of NaHCO3 and DIDS, cAMP concentrations were significantly lower than the values obtained from sperm incubated in complete medium. Protein tyrosine phosphorylation has also been shown to be highly correlated with the onset of capacitation in many species. During the first hour of capacitation, an increase in protein tyrosine phosphorylation was observed in complete medium. In the absence of NaHCO3, the increase in protein tyrosine phosphorylation was delayed for 45 min, and this delay was overcome by the addition of dbcAMP and IBMX. The induction of the acrosome reaction by calcium ionophore A23187 in NaHCO3-free medium was delayed 2 h, as compared with control medium. This delay was not observed in the presence of dbcAMP and IBMX. Taken together, these results suggest that a cAMP pathway may mediate the role of NaHCO3 in the capacitation of hamster spermatozoa and that protein tyrosine phosphorylation is necessary but not sufficient for complete capacitation. PMID- 10377036 TI - Differential activity of diethylstilbestrol versus estradiol as neonatal endocrine disruptors in the female hamster (Mesocricetus auratus) reproductive tract. AB - The synthetic estrogen diethylstilbestrol (DES) is a potent neonatal endocrine disruptor in the hamster. To test the specificity of this phenomenon, newborn animals were treated with 100 microgram of either DES or the natural estrogen, estradiol-17beta (E2). Of the two, neonatal DES exposure caused greater morphological disruption throughout the female reproductive tract in prepubertal animals and in adults that either retained their ovaries or were ovariectomized and then given the same levels of chronic E2 stimulation. In the uterus, a characteristic histopathological profile, including enhancement of both hyperplastic and apoptotic activity, was initiated prepubertally and exclusively in the endometrial epithelial cell compartment from the neonatally DES-treated animals and then was promoted by E2 stimulation during adulthood. Interestingly, apoptotic activity was not detected in an area of endometrial epithelium that progressed to the neoplastic state in a DES-exposed animal. Lastly, chronic estrogen induction of lactoferrin was also restricted to the DES-exposed endometrium. We conclude that 1) DES is more active than E2 as a perinatal endocrine disruptor in the hamster and 2) this experimental system should be generally useful as a means to screen compounds for such activity and then probe their mechanism of action. PMID- 10377035 TI - Regulation of monocyte chemotactic protein-1 expression in human endometrial stromal cells by estrogen and progesterone. AB - There is a cyclicity in the number of endometrial macrophages that is most likely secondary to changes in steroid hormone levels. One cytokine that controls macrophage migration is monocyte chemotactic protein-1 (MCP-1). In the endometrium, highest levels of MCP-1 are detected perimenstrually, when estrogen levels are low; however, when estrogen levels are high (around the time of ovulation), MCP-1 levels are lowest. We hypothesized that sex steroids may be involved in the regulation of macrophage migration by regulating MCP-1 expression. We investigated the regulation of MCP-1 expression in human endometrial stromal cells by estradiol 17beta (E2) and progestins. We found that MCP-1 mRNA levels decreased in response to E2 (5 x 10(-8) M), with biphasic nadirs at 8 h and 24 h. MCP-1 protein production was also inhibited by E2 in a concentration-dependent manner. Tamoxifen, an anti-estrogen, alone (10(-7) M) did not affect MCP-1 expression, but it reversed the E2-induced inhibition up to 80%. Progesterone (10(-7) M) alone slightly decreased MCP-1 levels, and the combination of E2 and progesterone further decreased them, but that decrease was not different from that observed using E2 treatment alone. In summary, we found that E2 inhibits MCP-1 expression in endometrial stromal cells, and we speculate that E2 may control endometrial macrophage migration by regulating MCP-1 expression. PMID- 10377037 TI - Switching maternal dietary intake at the end of the first trimester has profound effects on placental development and fetal growth in adolescent ewes carrying singleton fetuses. AB - The aim was to investigate whether placental growth and hence pregnancy outcome could be altered by switching adolescent dams from a high to a moderate nutrient intake, and vice-versa, at the end of the first trimester. Embryos recovered from adult ewes inseminated by a single sire were transferred in singleton to peripubertal adolescents. After transfer, adolescent ewes were offered a high (H, n = 33) or moderate (M, n = 32) level of a diet calculated to promote rapid or moderate maternal growth rates, respectively. At Day 50 of gestation, half the ewes had their dietary intakes switched, yielding 4 treatment groups: HH, MM, HM, and MH. A subset of ewes were killed at Day 104 of gestation to determine maternal body composition in relation to growth of the products of conception. Maternal body composition measurements revealed that the higher live weight in the high-intake dams was predominantly due to an increase in body fat deposition, with a less pronounced increase in body protein. At Day 104, HH and MH groups (high intake during second trimester) compared with MM and HM groups (moderate intake during second trimester) had a lower (p < 0.002) total fetal cotyledon weight; but fetal weight, conformation, and individual organ weights were not significantly influenced by maternal dietary intake. In ewes delivering live young at term, a high plane of nutrition from the end of the first trimester (HH and MH groups) compared with moderate levels (MM and HM groups) was associated with a reduction in gestation length (p < 0.009), total placental weight (p < 0.002), total fetal cotyledon weight (p < 0.001), and mean fetal cotyledon weight per placenta (p < 0.001). Fetal cotyledon number was dependent on maternal dietary intake during the first trimester only and was lower (p < 0.007) in HH and HM ewes compared to MM and MH ewes. The inhibition of fetal cotyledon growth in HH and MH groups was associated with a major decrease (p < 0.001) in lamb birth weight at term relative to the MM and HM groups. Thus, reducing maternal dietary intake from a high to a moderate level at the end of the first trimester stimulates placental growth and enhances pregnancy outcome, and increasing maternal dietary intake at this time point has a deleterious effect on placental development and fetal growth. PMID- 10377038 TI - Effect of myosin light chain kinase, protein kinase A, and protein kinase C inhibition on porcine oocyte activation. AB - Previous studies have shown that exposure to broad-spectrum protein kinase inhibitors results in parthenogenetic activation of metaphase II arrested porcine oocytes. The objective of this study was to determine the effect of inhibitors of myosin light chain kinase and other protein kinases on pronuclear development, dephosphorylation of a 25-kDa protein, and cortical granule exocytosis. Metaphase II arrested oocytes were obtained by in vitro maturation. Cumulus-free oocytes were cultured with specific inhibitors in modified Whitten's medium for 24 h. Treatment with inhibitors that should inhibit myosin light chain kinase--HA100 (250 microM), Wortmannin (1 microM), and a combination of Wortmannin (1 microM), KT5720 (75 nM), and Iso-H7 (50 microM)--resulted in significantly higher pronuclear development (74.0%, 18.0%, and 35.0%, respectively) than in the negative control, H7 (10 microM; 2.0-12.4% depending upon the replication). Treatment with HA100 (250 microM) resulted in the dephosphorylation of the 25-kDa protein to a 22-kDa protein in 80.0% (n = 10) of oocytes exposed. However, Wortmannin (1 microM; n = 17), KT5720 (75 nM; n = 16), and Iso-H7 (50 microM; n = 19) treatment individually and in combination (n = 19) did not result in significant (p < 0.05; n = 19) dephosphorylation over the negative control, H7 (10 microM; n = 19). HA100 treatment resulted in significant cortical granule exocytosis when evaluated by laser confocal microscopy. In addition, protein kinase assays revealed lower myosin light chain kinase activity in electroactivated oocytes (p < 0.05) and protein kinase inhibitor-treated oocytes (p < 0.05) than in negative controls, nonelectroactivated oocytes, and H7 (10 microM)-treated oocytes. Treatment with HA100 (250 microM) resulted in pronuclear formation, dephosphorylation of the 25-kDa protein, and some release of cortical granules. These observations suggest that inhibition of myosin light chain kinase, protein kinase A, and protein kinase C results in activation of porcine oocytes. PMID- 10377039 TI - Transcription factors Ets1, Ets2, and Elf1 exhibit differential localization in human endometrium across the menstrual cycle and alternate isoforms in cultured endometrial cells. AB - To better understand the transcriptional regulation of human endometrial remodeling, the localization of three members of the Ets family of transcription factors was examined at different stages of the menstrual cycle. Elf1 was found by immunohistochemistry to be predominantly localized to the glandular epithelium. In contrast, Ets1 and Ets2 were found at lower intensities in both glandular epithelial and stromal cells. Low expression during the menstrual phase of the cycle, and high expression and intensity of staining in decidualized stromal cells of the late secretory phase were common to Ets1, Ets2, and Elf1. These localization patterns were confirmed in cultured human endometrial stromal and epithelial cells by Western blotting, which also demonstrated different isoforms and phosphorylation products of Ets1 and Ets2 in the two cell types. This study has shown for the first time that members of the Ets family of transcription factors, previously found predominantly during development and in hematopoietic cells, are expressed in the human endometrium and display cell and cycle-stage specificity. Expression of Elf1 predominantly in the glandular epithelium may indicate that Elf1 plays a unique role in epithelium-specific gene regulation in the endometrium. PMID- 10377040 TI - Persistent dominant follicle alters pattern of oviductal secretory proteins from cows at estrus. AB - The experimental objective was to compare synthesis of oviductal secretory proteins of dairy cows bearing a persistent dominant follicle (PDF) versus a fresh dominant follicle (FDF) at estrus. On Day 7 after synchronized estrus (Day 0), cows received an intravaginal progesterone device and injection of prostaglandin F2alpha (PGF2alpha). On Day 9, cows received an injection of a GnRH agonist (FDF group; n = 3) or received no injection (PDF group, n = 3). On Day 16, all cows received PGF2alpha, and progesterone devices were removed. At slaughter on Day 18 or Day 19, oviducts ipsilateral and contralateral to the dominant follicle were divided into infundibulum, ampulla, and isthmus regions. Explants from oviductal regions were cultured in minimal essential medium supplemented with [3H]leucine for 24 h. Two-dimensional fluorographs of proteins in conditioned media were analyzed by densitometry. Rate of incorporation of [3H]leucine into macromolecules was greater in the infundibulum, ampulla, and isthmus of FDF cows (p < 0.01). Overall, intensities of radiolabeled secretory protein (P) 2 and P13 were greater for FDF than for PDF. In the ampulla, P14 was more intense for FDF while P7 was more intense for PDF. Abundance of P1 in the isthmus was greater for PDF cows. Across regions, P5, P6, P8, P9, and P11 were more intense for PDF than for FDF in the ipsilateral side. In the contralateral side, P19 was more intense for PDF than for FDF, whereas P6, P8, P9, and P11 were more intense for FDF. Differences in biosynthetic activity and in secreted oviductal proteins from cows bearing a PDF may contribute to the decrease in fertility associated with a PDF. PMID- 10377041 TI - Sensitivity of domestic cat (Felis catus) sperm from normospermic versus teratospermic donors to cold-induced acrosomal damage. AB - Freeze-thawing cat sperm in cryoprotectant results in extensive membrane damage. To determine whether cooling alone influences sperm structure and viability, we compared the effect of cooling rate on sperm from normospermic (N; > 60% normal sperm per ejaculate) and teratospermic (T; < 40% normal sperm per ejaculate) domestic cats. Electroejaculates were divided into raw or washed (Ham's F-10 + 5% fetal calf serum) aliquots, with the latter resuspended in Ham's F-10 medium or Platz Diluent Variant Filtered without glycerol (20% egg yolk, 11% lactose). Aliquots were 1) maintained at 25 degrees C (no cooling; control), 2) cooled to 5 degrees C in a commercial refrigerator for 30 min (rapid cooling; approximately 4 degrees C/min), 3) placed in an ice slush at 0 degrees C for 10 min (ultrarapid cooling; approximately 14 degrees C/min), or 4) cooled to 0 degrees C at 0.5 degrees C/min in a programmable alcohol bath (slow cooling); and aliquots were removed every 4 degrees C. All samples then were warmed to 25 degrees C and evaluated for percentage sperm motility and the proportion of intact acrosomes using a fluorescein-conjugated peanut agglutinin stain. In both cat populations, sperm percentage motility remained unaffected (p > 0.05) immediately after exposure to low temperatures and after warming to 25 degrees C. However, the proportion of spermatozoa with intact acrosomes declined (p < 0.05) after rapid cooling ( approximately 4 degrees C/min) to 5 degrees C (N, 65.6%; T, 27.5%) or ultrarapid cooling ( approximately 14 degrees C/min) to 0 degrees C (N, 62.1%; T, 23.0%) in comparison to the control value (N, 81.5%; T, 77.5%). Transmission electron microscopy of cooled sperm revealed extensive damage to acrosomal membranes. In contrast, slow cooling (0.5 degrees C/min) to 5 degrees C maintained (p > 0.05) a high proportion of spermatozoa with intact acrosomes (N, 75.5%; T, 68.3%), which also remained similar (p > 0.05) between cat populations (N, 64.7%; T, 56.8%) through continued cooling to 0 degrees C. Results demonstrate that 1) rapid cooling of domestic cat sperm induces significant acrosomal damage without altering sperm motility, 2) spermatozoa from teratospermic males are more susceptible to cold-induced acrosomal damage than normospermic counterparts, and 3) reducing the rate of initial cooling markedly decreases sperm structural damage. PMID- 10377042 TI - Functional capacity of fetal zone cells of the baboon fetal adrenal gland: a major source of alpha-inhibin. AB - We have shown that ACTH receptor mRNA expression and steroidogenesis were increased in the transitional zone and decreased in the fetal zone of the baboon fetal adrenal in the second half of gestation. Thus, we proposed that there is a divergence in ACTH receptor-mediated zone-specific steroidogenesis within the fetal adrenal during mid to late gestation. We have also demonstrated that fetal serum alpha-inhibin levels decline with advancing development. It is possible, therefore, that the alpha subunit of inhibin provides a good marker of fetal zone cellular function and that the changes in circulating fetal alpha-inhibin with advancing pregnancy reflect ontogenetic changes in fetal adrenal cortical zone specific cell function. However, it remains to be determined whether the fetal adrenal is a major source of circulating alpha-inhibin in the fetus and whether alpha-inhibin is expressed in the fetal, definitive, and/or transitional zones. Therefore, the current study compared fetal serum alpha-inhibin levels with immunocytochemical localization of alpha-inhibin in baboon fetal adrenals obtained on Days 60 (early), 100 (mid), and 165 or 182 (late) of gestation (term averages Day 184) from animals untreated or treated with betamethasone, which we previously demonstrated suppressed fetal pituitary ACTH and adrenal weight. Fetal serum alpha-inhibin levels (mean +/- SE) were greater (p < 0.05) at mid (5863 +/- 730 microliter eq/ml) than at late (3246 +/- 379) gestation and were reduced (p < 0. 05) by betamethasone. The inhibin alpha subunit was expressed in abundant quantities in the fetal adrenal cortex, but not in medulla, throughout gestation. At mid and late gestation, alpha-inhibin was expressed throughout the fetal adrenal cortex but most intensely in the innermost area of fetal zone cells. By late gestation, the fetal adrenal exhibited a gradient of alpha-inhibin expression. Thus, the outermost definitive zone cells were devoid of alpha inhibin, the transitional zone exhibited a relatively low alpha-inhibin content, and fetal zone cells continued to exhibit extensive expression of alpha-inhibin. Betamethasone diminished the intensity of alpha-inhibin expression throughout the fetal adrenal cortex. These results indicate that the fetal adrenal fetal zone is a significant source of circulating alpha-inhibin in the baboon fetus and that alpha-inhibin provides a good marker to study the developmental regulation of fetal zone-specific adrenocortical function. PMID- 10377043 TI - Diverse effects of tyrosine kinase inhibitors on follicle-stimulating hormone stimulated estradiol and progesterone production from rat granulosa cells in serum-containing medium and serum-free medium containing epidermal growth factor. AB - Epidermal growth factor (EGF) has been shown to influence FSH-stimulated estradiol (E2) and progesterone (P4) production from granulosa cells. RG 50810, a tyrosine kinase inhibitor (TKI), has previously been shown to inhibit the EGF receptor tyrosine kinase. RG 50810 has also been shown to inhibit FSH-stimulated increases in mRNA for steroidogenic enzymes, implying a functional role of tyrosine kinases in FSH action in granulosa cells. However, inhibition of FSH stimulated steroidogenesis by TKIs has not been evaluated in connection with the effects of EGF in granulosa cells. In the present studies, FSH-stimulated E2 production was inhibited similarly by inhibitors of protein kinase A (H-89) and protein kinase C (calphostin C) and by TKIs, and none of the inhibitors were capable of reversing the EGF-induced inhibition of FSH-stimulated E2 production. FSH-stimulated P4 production was enhanced dramatically in serum-containing medium with concentrations of TKI that were near previously reported IC50s. The enhancing effect of TKIs was less evident in serum-free medium. Addition of EGF to serum-free medium enhanced FSH-stimulated P4 production, and the TKIs reversed EGF-enhanced P4 production, but in a manner similar to that of protein kinase A inhibitor H-89. Compared to results in serum-free medium, the potency of RG 50810 and genistein to inhibit the effects of EGF on P4 production was 3- to 8-fold greater relative to H-89. These studies have demonstrated that TKIs RG 50810 and genistein selectively inhibit the effects of EGF on FSH-stimulated P4 production in granulosa cell cultures. In contrast, these studies have demonstrated nonselective inhibition of FSH-stimulated E2 and P4 production by TKIs in serum free medium, in which it is not clear which enzyme system is affected by the compounds tested. PMID- 10377044 TI - Improvement of spermatogenesis in adult cryptorchid rat testis by intratesticular infusion of lactate. AB - In order to test the hypothesis that a lack of energy could be a cause of germ cell death at high temperatures, cryptorchid rats testes were infused with lactate, delivered by osmotic pumps over 3-15 days. In cryptorchid testes, the spermatids and spermatocytes were lost between 3 and 8 days. In cryptorchid testes supplemented with lactate, elongated spermatids persisted in a few seminiferous tubules at Day 15. Elimination of round spermatids occurred progressively between 3 and 15 days, mostly at stage VIII. The loss of spermatocytes increased after 8 days, and 30% of seminiferous tubules still contained meiotic or meiotic plus spermiogenetic cells at Day 15. After 8 days, the chromatin of step 8 round spermatids was abnormal and nuclear elongation did not commence. The Sertoli cell cytoplasm that was retracted toward the basal compartment of the seminiferous epithelium could not hold the germ cells of the adluminal compartment. Therefore, attachment of germ cells to Sertoli cells and the supply of lactate seem necessary for the development of germ cells at high temperatures. The improvement in spermatogenesis in cryptorchid supplemented testes for several days is a new finding. PMID- 10377045 TI - Hormonal regulation of natriuretic peptide system during induced ovarian follicular development in the rat. AB - All components of the natriuretic peptide (NP) system have been found in the ovary. The purpose of this study was to determine the hormonal regulation of the NP system during follicular growth and ovulation induced by gonadotropins eCG and hCG. Ovarian membrane binding, before and after treatment, revealed the presence of guanylyl cyclase-type receptors exclusively. Equine CG treatment increased Bmax from 225 +/- 50 fmol/mg protein in control animals to 354 +/- 51 fmol/mg protein, and additional hCG treatment increased it further to 492 +/- 130 fmol/mg protein (p < 0.05), without changing receptor affinity. The increased binding was consistent with increased ability of atrial natriuretic peptide (ANP) to activate guanylyl cyclase in the ovarian cells obtained from hormone-treated animals. In confirmation, autoradiography of 125I-tyroCNP and 125I-ANP binding to the rat ovary showed that both guanylyl cyclase GC-A and GC-B receptor subtypes are localized to the granulosa cells of antral follicles. Quantitative analysis of GC A and GC-B receptors by reverse transcription-polymerase chain reaction showed that the expression level of both receptors started to increase at 2 h and reached maximal levels at 6 h following eCG treatment. Increased levels of GC-B mRNA were also observed 12 h after eCG injection. At 24 and 48 h the receptor levels were below basal. Stimulation of NP receptors by eCG was paralleled by activation of both ovarian ANP and C-type natriuretic peptide (CNP) gene expression. ANP mRNA increased as early as 1 h after eCG injection and remained elevated up to 6 h. CNP mRNA increased at 2 h after eCG injection, peaked (5 fold) at 6 h, and remained elevated 48 h later, a stage at which follicular maturation continues. Incubation of ovaries with ANP significantly decreased eCG induced estradiol level, indicating the functionality of the ovarian NP system. These results implicate the NP system in the induction and maintenance of fluid balance in the rapidly developing ovarian follicle. PMID- 10377046 TI - Abnormal estrous cyclicity after disruption of endothelial and inducible nitric oxide synthase in mice. AB - The roles of nitric oxide (NO) and nitric oxide synthase (NOS) in reproduction were studied by examining the estrous cycle of wild-type (WT) mice, inducible NOS (iNOS)-, and endothelial NOS (eNOS)-knockout mice. We observed an average estrous cycle of 4.8 +/- 0.2 days in WT mice. While we observed no significant influence of iNOS deficiency on cycle length, eNOS-knockout females showed a significantly longer estrous cycle (6.6 +/- 0.6 days; p < 0.03) than WT females, due to an extension of diestrus (p < 0.03). There was no influence of iNOS deficiency on ovulation rate compared with that in WT females; however, eNOS-knockout mice showed a significant reduction (p < 0.05) in ovulatory efficiency relative to WT or iNOS-knockout females. In contrast to WT females, in which the highest level of estradiol (E2) was observed at 1500 h of proestrus, iNOS-knockout females reached a peak of E2 at 1830 h of proestrus. In eNOS-knockout females, the peak of E2 occurred at 1830 h, as in iNOS-knockout mice; however, E2 levels were 5 fold and 3-fold higher (p < 0.05) than levels observed in WT and iNOS-knockout females, respectively. There was no effect of genotype on the plasma LH concentrations at proestrus. On the first day of diestrus, eNOS-knockout females showed significantly higher plasma E2 and progesterone levels (p < 0.05) relative to WT and iNOS-knockout females. The dysfunction in cyclicity, ovulation rate, ovarian morphology, and steroidogenesis in eNOS-knockout female mice strongly supports the concept that eNOS/NO plays critical roles in ovulation and follicular development. PMID- 10377048 TI - Circannual variations in intraovarian oocyte but not epididymal sperm quality in the domestic Cat. AB - Ovaries and testes were collected throughout the year from domestic cats spayed and neutered at local veterinary clinics. Fresh oocytes recovered from minced ovaries were subjected to in vitro maturation and then stained to determine stage of maturation or were inseminated with conspecific sperm. The cauda and corpus regions of each epididymis were dissected into pieces and placed in medium; 30 min later, the epididymal tissue was removed, the medium centrifuged, and the sperm pellet resuspended. Samples were assessed for total sperm count and sperm motility traits, morphology, acrosomal integrity, and ability to penetrate cat oocytes in vitro. Fewer excellent (grade I) oocytes were recovered per ovarian pair during September-November (mean +/- SEM, 19.2 +/- 2.1%) than during January July (36.8 +/- 3.6%, p < 0.05), while the remaining months had intermediate percentages of grade I oocytes (p > 0.05). A high percentage of oocytes recovered from November-April completed nuclear maturation (64.3 +/- 6.8%), which was different (p < 0.05) from the values for May-July (32.2 +/- 3.8%) and August October (10.4 +/- 2.9%). Percentage of oocytes with bound sperm was lowest (p < 0.01) in September and October (32.0 +/- 3.1%) compared to February and March (91.4 +/- 1.7%). Percentage of oocytes with sperm within the perivitelline space was highest (p < 0.05) in May-August (33.8 +/- 4.6%) compared to all other months. In contrast, the period of highest (p < 0.01) fertilization (i.e., >/= 4 cell embryo formation) was March-April (51.7 +/- 3.1%) as compared to May-July (17.2 +/- 1.8%) or November-January (12.4 +/- 2.6%). Negligible numbers of oocytes recovered during August-October developed beyond the 2-cell stage (1.1 +/ 0.3%). Blastocyst development from cleaved embryos was highest during February April (44.3 +/- 2.3%) and lowest during August-October (0.6 +/- 0.1%; p < 0.01). Sperm recovered from the epididymides throughout the year did not differ (p > 0.05) in concentration or in any of the motility, structural, or functional variables evaluated. In summary, cat oocyte nuclear maturation in vitro is depressed during August-October, and the ability to form cleaved embryos remains low even when the capacity to achieve nuclear maturation is relatively high (November-January and May-July). In contrast, male cats are capable of consistently producing viable, progressively motile sperm throughout the year. PMID- 10377047 TI - Effects of cryopreservation procedures on the cytology and fertilization rate of in vitro-matured bovine oocytes. AB - The survival and developmental capacity of bovine oocytes after cryopreservation are greatly impaired, possibly due to organelle damage caused by freezing procedures. Distributions of chromosomes, microtubules, and microfilaments in bovine oocytes matured in vitro were examined after cooling, ethylene glycol (EG) exposure, or freezing. Oocytes were incubated after treatment for 20 min or 1 or 3 h, fixed, and evaluated using specific fluorescent probes. Abnormal cytological features increased over control levels after cooling or EG exposure and rewarming. Changes observed in oocytes during prefreezing manipulations included chromosome dispersal and clumping, microtubule depolymerization and alteration of spindle structure, and formation of craters and discontinuity in cytoskeletal actin staining. Freezing also led to an increase in the occurrence of cytological abnormalities. Less than 31% of frozen-thawed oocytes contained a normal chromosome arrangement 3 h postthaw (versus 90% of controls). Only 7-14% of frozen-thawed oocytes had normal spindles (versus 59-71% of controls). Normal distribution of filamentous actin was observed in less than 30% of oocytes postthaw (versus 62-89% of controls). These results indicate that the steps in a conventional freezing procedure cause irreversible alterations in multiple cytological components of bovine oocytes, demonstrating the need for improved strategies for preventing cellular damage during cryopreservation procedures. PMID- 10377049 TI - Production of lysophosphatidic acids by lysophospholipase D in human follicular fluids of In vitro fertilization patients. AB - Lysophosphatidic acids (LPAs) are known to be normal constituents of mammalian serum, and they mimic some biological effects of the serum. We previously reported that lysophospholipase D (LPLD) was involved in the accumulation of LPAs in incubated rat plasma and serum. In this study we detected, by gas-liquid chromatography, various molecular species of LPA in follicular fluids collected from women programmed for in vitro fertilization. When the follicular fluid was incubated at 37 degrees C for 48 h, persistent increases in the amounts of LPAs were observed concomitant with decreases in the amounts of the corresponding lysophosphatidylcholines (LPCs), although the concentrations of saturated LPCs increased in the first 6 h of incubation. These results suggest that human follicular fluid has LPLD activity, and this was confirmed by experiments with follicular fluids mixed with an exogenous radioactive LPC. The LPLD showed preference for unsaturated over saturated LPCs, similar to plasma LPLD, indicating that it originated from the circulation. PMID- 10377050 TI - Imprinting by neonatal sex steroids on the structure and function of the mature mouse prostate. AB - Perinatal sex-steroid exposure may result in permanent modifications in the structure and function of the prostate gland. The mechanism of such long-range alterations in hormonal sensitivity is not known. This study aimed to define the molecular requirements for neonatal sex-steroid imprinting and to investigate whether combined administration of neonatal androgens and estrogens had synergistic effects upon the mature mouse prostate. Since the interaction between endogenous and exogenous sex steroids in normal mice makes it difficult to dissociate direct from indirect effects, we used the hypogonadal (hpg) mouse, characterized by congenital androgen deficiency yet still fully responsive to exogenous androgens. Newborn mice (Days 1-2) were administered a single s.c. injection of androgens alone or in combination with an estrogen followed by testosterone-induced maximal prostate growth at maturity. The final effects were determined in 7-wk-old mice through study of ductal architecture in microdissected ventral prostates (VP) and quantitation of volume densities and diameters of prostate tissue components. A single neonatal dose of androgens, but not of estrogen, increased branching morphogenesis and VP weights at adulthood. These effects did not differ significantly between various androgens; in addition, combined androgen and estrogen treatment failed to demonstrate any synergistic effects on the prostate. We conclude that neonatal androgens induce long-range effects upon the mature VP structure as well as its secretory function and that this imprinting occurs via the androgen receptor without requiring aromatization of androgens. However, these conclusions, based on a specific treatment protocol, are confined only to the distal segment of VP, and effects of neonatal sex-steroid exposure in other regions or lobes of VP may differ. PMID- 10377051 TI - Humoral immune response to equine chorionic gonadotropin in ewes: association with major histocompatibility complex and interference with subsequent fertility. AB - In dairy ewes, the use of eCG as a convenient hormone for the induction of ovulation is necessary for out-of-season breeding and artificial insemination (AI). In this report we show the presence of anti-eCG antibodies in plasma of treated ewes. The major histocompatibility complex (MHC) was involved in the individual variability of the humoral immune responses to eCG. We found significant associations between the anti-eCG response phenotype and some MHC class II alleles. The low immune response phenotype was associated with one MHC class II allele only in Lacaune ewes, and the high immune response phenotype was associated with one MHC class II allele both in Manech and in Lacaune ewes. In herds, the impact of residual anti-eCG antibodies on subsequent fertility after AI seems minimal because of an indirect elimination of high-responder ewes from AI breeding. Therefore, the true magnitude of the association between residual anti-eCG antibody concentration and fertility has been underestimated. An additional experiment without any high-responder female elimination showed a significant correlation between high residual antibody concentrations and lower lambing rate after AI at a fixed time, possibly because of a delayed preovulatory LH surge. The results suggest that anti-eCG antibody concentration is one risk factor for infertility after AI. PMID- 10377052 TI - Insulin-like growth factor-I, insulin-like growth factor-binding proteins, and gonadotropins in the hypothalamic-pituitary axis and serum of nutrient-restricted ewes. AB - Body condition scores (BCS) of ovariectomized estradiol-treated ewes were controlled to examine effects of suboptimum BCS on insulin-like growth factor (IGF)-I, IGF-binding proteins (IGFBPs), and LH in the anterior pituitary gland, hypophyseal stalk-median eminence (SME), and circulation. Serum LH increased in ewes with BCS (1 = emaciated, 9 = obese) > 3 (HIGH-BCS), but not in ewes with BCS cIL-1RA) expression, 2) localize the relevant transcripts to the granulosa cell, 3) disclose peak expression at the time of ovulation, and 4) establish IL-1 dependence. PMID- 10377061 TI - Complementary deoxyribonucleic acid cloning and tissue expression of BSP-A3 and BSP-30-kDa: phosphatidylcholine and heparin-binding proteins of bovine seminal plasma. AB - BSP-A1, BSP-A2, BSP-A3, and BSP-30-kDa are four major proteins of bovine seminal plasma (BSP protein family). These heparin- and phosphatidylcholine-binding proteins potentiate the capacitation of spermatozoa. Here we determined the complete sequences of the two cDNAs coding for the BSP-A3 and BSP-30-kDa proteins. Degenerate oligonucleotides designed on the basis of the primary sequences of the proteins were used as primers in reverse transcription polymerase chain reaction, with cDNA preparations of bovine seminal vesicles as templates, to amplify an internal fragment of each BSP cDNA. Specific oligonucleotides designed on the basis of these partial cDNA sequences were used to clone the two complete cDNAs by using the 3' rapid amplification of cDNA ends (RACE) and 5' RACE methods. We also verified the expression of all members of the bovine BSP protein family in several adult bovine tissues by RNase protection assays. The results indicated that each BSP protein mRNA is expressed only in seminal vesicles and in the ampullae. Homologous genes were detected in human, rat, hamster, and rabbit genomic DNA, using high-stringency Southern hybridization with a specific BSP-30-kDa cDNA probe. PMID- 10377062 TI - Production of plasminogen activators (PAs) in bovine cumulus-oocyte complexes during maturation in vitro: effects of epidermal growth factor on production of PAs in oocytes and cumulus cells. AB - We examined whether plasminogen activators (PAs) are produced by bovine cumulus oocyte complexes (COCs) during maturation in vitro. The effects of epidermal growth factor (EGF) on production of PAs in oocytes and cumulus cells were also examined. When COCs were cultured for 24 h with 30 ng/ml EGF, three plasminogen dependent lytic zones (58.5 +/- 3.5 kDa, 79.0 +/- 3.0 kDa, and 113.5 +/- 6.5 kDa) were observed. Addition of amiloride, a competitive inhibitor of urokinase-type PA (uPA), to the zymogram eliminated the activity of the 58.5 +/- 3.5-kDa zone, suggesting that this band is a uPA. However, since the activity of the remaining two bands was not eliminated, it was suggested that the 79.0 +/- 3.0-kDa band is a tissue-type PA (tPA) and the 113.5 +/- 6.5-kDa band is possibly a tPA-PA inhibitor (tPA-PAI) complex. In COCs before culture, however, no activity of PAs was detected. At 6 h of culture, the same level of uPA activity was detected in COCs cultured both in the absence and in the presence of EGF. The uPA activity was increased at 12 h of culture but without further increase at 24 h of culture, with higher activity in the presence than in the absence of EGF. The activity of tPA and tPA-PAI was first detected at 24 h of culture in the absence of EGF. In the presence of EGF, however, some activity of tPA-PAI was detected at 12 h of culture. At 24 h of culture, the activity of all PAs was detected in cumulus cells, but only uPA activity was detected in oocytes, with higher activity in the presence than in the absence of EGF. The uPA activity in oocytes was not detected when they were cultured without cumulus cells in either the presence or absence of EGF, although cumulus expansion was stimulated by EGF, exhibiting a time course similar to that observed in PA production. These results suggest that uPA, tPA, and tPA-PAI are all produced by bovine COCs, but only uPA by oocytes, during maturation in vitro. However, cumulus cells play an essential role or roles in the production of uPA by oocytes, and EGF enhances the roles of cumulus cells. PMID- 10377063 TI - Mice carrying two t haplotypes: sperm populations with reduced Zona pellucida binding are deficient in capacitation. AB - Capacitation is the unique process by which mammalian sperm become capable of undergoing the acrosome reaction (AR). An approach to studying sperm capacitation is to identify mutations altering this process. Male mice carrying two t haplotypes are sterile, with poor sperm motility, reduced zona pellucida binding, and an inability to penetrate zona-free oocytes. The objective of this study was to examine sperm capacitation and its potential relationship to zona pellucida binding in mice of the same genetic strain carrying none, one, or two t haplotypes. Sperm capacitation was assessed by the B pattern of staining by chlortetracycline (CTC) and by the ability of sperm to undergo the lysophosphatidylcholine (LPC)-induced AR. The CTC assay demonstrated that sperm capacitation from t/+ mice was similar to that from +/+ mice, but sperm from t/t mice were deficient. LPC induced the AR of capacitated sperm, but not noncapacitated sperm, in a concentration-dependent manner. Sperm from t/t mice were also deficient in the LPC-induced AR. Thus, by two independent assays, sperm from t/t mice were shown to be deficient in capacitation. To determine whether a deficiency in capacitation could influence zona binding, the ability of capacitated versus noncapacitated sperm to bind to the zona pellucida was tested. The mean numbers of sperm bound per oocyte were significantly greater for capacitated sperm than for noncapacitated sperm. These results suggest that the deficient capacitation of sperm from t/t mice could be responsible for, or at least contribute to, their reduced ability to bind to the zona pellucida. PMID- 10377064 TI - Expression of the interferon tau inducible ubiquitin cross-reactive protein in the ovine uterus. AB - Ubiquitin cross-reactive protein (UCRP) is a 17-kDa protein that shows cross reactivity with ubiquitin antisera and retains the carboxyl-terminal Leu-Arg-Gly Gly amino acid sequence of ubiquitin that ligates to, and directs degradation of, cytosolic proteins. It has been reported that bovine endometrial UCRP is synthesized and secreted in response to conceptus-derived interferon-tau (IFNtau). In the present studies, UCRP mRNA and protein were detected in ovine endometrium. Ovine UCRP mRNA was detectable on Day 13, peaked at Day 15, and remained high through Day 19 of pregnancy. The UCRP mRNA was localized to the luminal epithelium (LE), stromal cells (ST) immediately beneath the LE, and shallow glandular epithelium (GE) on Day 13, but it extended to the deep GE, deep ST, and myometrium of uterine tissues by Day 15 of pregnancy. Western blotting revealed induction of UCRP in the endometrial extracts from pregnant, but not cyclic, ewes. Ovine UCRP was also detected in uterine flushings from Days 15 and 17 of pregnancy and immunoprecipitated from Day 17 pregnant endometrial explant conditioned medium. Treatment of immortalized ovine LE cells with recombinant ovine (ro) IFNtau induced cytosolic expression of UCRP, and intrauterine injection of roIFNtau into ovariectomized cyclic ewes induced endometrial expression of UCRP mRNA. These results are the first to describe temporal and spatial alterations in the cellular localization of UCRP in the ruminant uterus. Collectively, UCRP is synthesized and secreted by the ovine endometrium in response to IFNtau during early pregnancy. Because UCRP is present in the uterus and uterine flushings, it may regulate endometrial proteins associated with establishment and maintenance of early pregnancy in ruminants. PMID- 10377065 TI - Mitogen-activated protein kinase activation by stimulation with thyrotropin releasing hormone in rat pituitary GH3 cells. AB - We examined whether mitogen-activated protein (MAP) kinase is activated by thyrotropin-releasing hormone (TRH) in GH3 cells, and whether MAP kinase activation is involved in secretion of prolactin from these cells. Protein kinase inhibitors--such as PD098059, calphostin C, and genistein--and removal of extracellular Ca2+ inhibited MAP kinase activation by TRH. A cAMP analogue activated MAP kinase in these cells. Effects of cAMP on MAP kinase activation were inhibited by PD098059. TRH-induced prolactin secretion was not inhibited by levels of PD098059 sufficient to i activation but was inhibited by wortmannin (1 microM) and KN93. Treatment of GH3 cells with either TRH or cAMP significantly inhibited DNA synthesis and induced morphological changes. The effects stimulated by TRH were reversed by PD098059 treatment, but the same effects stimulated by cAMP were not. Treatment of GH3 cells with TRH for 48 h significantly increased the prolactin content in GH3 cells and decreased growth hormone content. The increase in prolactin was completely abolished by PD098059, but the decrease in growth hormone was not. These results suggest that TRH-induced MAP kinase activation is involved in prolactin synthesis and differentiation of GH3 cells, but not in prolactin secretion. PMID- 10377067 TI - Circulation online only : june 22, 1999 PMID- 10377066 TI - Progesterone regulation of epidermal growth factor receptor in rat decidua basalis during pregnancy. AB - Ovarian steroid hormones and epidermal growth factor (EGF) play important interactive roles in proliferation and decidualization of mesometrial stromal cells during pregnancy. This study determined the ontogeny of EGF receptor (EGF R) expression in the decidua basalis (DB) throughout pregnancy and its regulation by estrogen and progesterone (P4). DB were isolated from rats between Days 8-21 of pregnancy and prepared for immunohistochemistry or Western analysis. In one study, rats were ovariectomized (Ovx) on Day 8 or 9 and given estradiol-17beta, P4, or both. In another study, the antiprogestin, mifepristone (RU-486), was administered on Day 9. During normal pregnancy, total EGF-R (phosphorylated and unphosphorylated forms) increased from Day 8 to a maximum level on Days 10 and 12. Tyrosine-phosphorylated EGF-R (pEGF-R), the bioactive form, was also highest on Days 10 and 12. Both forms of receptor decreased to almost undetectable levels during DB regression on Days 17-21. Immunohistochemistry of DB from Ovx rats revealed that only P4 was able to maintain normal expression of EGF-R; RU-486 decreased EGF-R expression within 6 h, and by 24 h EGF-R and pEGF-R were 15% of the Day 10 control group levels. These findings show that EGF-R is a P4-dependent protein associated with stromal cell proliferation and decidualization. PMID- 10377068 TI - Many cardiologists still unsure of electron beam computed tomography's benefit as a screening tool. PMID- 10377069 TI - Novel mechanism for endothelial dysfunction: dysregulation of dimethylarginine dimethylaminohydrolase. AB - BACKGROUND: Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of nitric oxide synthase (NOS). Plasma levels of ADMA are elevated in individuals with hypercholesterolemia or atherosclerosis. We postulated that reduced degradation of ADMA may play a role in the accumulation of ADMA in these individuals. Accordingly, we studied the effects of oxidized LDL (oxLDL) or tumor necrosis factor-alpha (TNF-alpha) on the accumulation of ADMA by transformed human umbilical vein endothelial cells (ECV304) and on the enzyme dimethylarginine dimethylaminohydrolase (DDAH), which degrades ADMA. METHODS AND RESULTS: ECV304 were incubated with or without native LDL (100 micrograms/mL), oxLDL (100 micrograms/mL), or TNF-alpha (250 U/mL) for 48 hours. The concentration of ADMA in the conditioned medium was determined by high performance liquid chromatography. Western blotting was performed to evaluate DDAH expression. We assayed DDAH activity by determining L-citrulline formation from ADMA. The addition of oxLDL or TNF-alpha to ECV304 significantly increased the level of ADMA in the conditioned medium. The effect of oxLDL or TNF-alpha was not due to a change in DDAH expression but rather to the reduction of DDAH activity. To determine whether dysregulation of DDAH also occurred in vivo, New Zealand White rabbits were fed normal chow or a high-cholesterol diet. Hypercholesterolemia significantly reduced aortic, renal, and hepatic DDAH activity. CONCLUSIONS: These results suggest that the endothelial vasodilator dysfunction observed in hypercholesterolemia may be due to reduced degradation of ADMA, the endogenous inhibitor of NOS. PMID- 10377070 TI - G894T polymorphism in the endothelial nitric oxide synthase gene is associated with an enhanced vascular responsiveness to phenylephrine. AB - BACKGROUND: Differences in vascular reactivity to phenylephrine (PE) responsiveness have been largely evidenced in patients undergoing cardiac surgery with cardiopulmonary bypass (CPB). Because nitric oxide (NO) strongly affects modulation of the vascular tone in response to vasopressor agents, we hypothesized that the G894T polymorphism of the endothelial NO synthase gene (eNOS) could be related to changes in the pressor response to PE. METHODS AND RESULTS: The protocol was performed in 68 patients undergoing coronary artery bypass grafting (n=33) or valve surgery (n=35) in whom mean arterial pressure decreased below 65 mm Hg during normothermic CPB. Under constant and nonpulsatile pump flow conditions (2 to 2.4 L. min-1. m-2), a PE dose-response curve was generated by the cumulative injection of individual doses of PE (25 to 500 micrograms). The G894T polymorphism of the eNOS gene was determined, and 3 groups were defined according to genotype (TT, GT, and GG). Groups were similar with regard to perioperative characteristics. The PE dose-dependent response was significantly higher in the allele 894T carriers (TT and GT) than in the homozygote GG group (P=0.02), independently of possible confounding variables. CONCLUSIONS: These results evidenced an enhanced responsiveness to alpha adrenergic stimulation in patients with the 894T allele in the eNOS gene. PMID- 10377071 TI - Cardiac-directed adenylyl cyclase expression improves heart function in murine cardiomyopathy. AB - BACKGROUND: We tested the hypothesis that increased cardiac myocyte adenylyl cyclase (AC) content increases cardiac function and response to catecholamines in cardiomyopathy. METHODS AND RESULTS: Transgenic mice with cardiac-directed expression of AC type VI (ACVI) were crossbred with mice with cardiomyopathy induced by cardiac-directed Gq expression. Gq mice had dilated left ventricles, reduced heart function, decreased cardiac responsiveness to catecholamine stimulation, and impaired beta-adrenergic receptor (betaAR)-dependent and AC dependent cAMP production. Gq/AC mice showed improved basal cardiac function in vivo (P=0.01) and ex vivo (P<0.0005). When stimulated through the betaAR, cardiac responsiveness was increased (P=0.02), and cardiac myocytes showed increased cAMP production in response to isoproterenol (P=0.03) and forskolin (P<0.0001). CONCLUSIONS: Increasing myocardial ACVI content in cardiomyopathy restores cAMP generating capacity and improves cardiac function and responsiveness to betaAR stimulation. PMID- 10377072 TI - Membrane type 1 matrix metalloproteinase expression in human atherosclerotic plaques: evidence for activation by proinflammatory mediators. AB - BACKGROUND: Matrix metalloproteinases (MMPs) are expressed in atherosclerotic plaques, where in their active form, they may contribute to vascular remodeling and plaque disruption. In this study, we tested the hypothesis that membrane type 1 MMP (MT1-MMP), a novel transmembrane MMP that activates pro-MMP-2 (gelatinase A), is expressed in human atherosclerotic plaques and that its expression is regulated by proinflammatory molecules. METHODS AND RESULTS: MT1-MMP expression was examined in normal and atherosclerotic human arteries by immunocytochemistry with specific antibodies. MT1-MMP expression in human saphenous vein-derived smooth muscle cells (SMCs) maintained in tissue culture was determined under basal conditions and in response to proinflammatory molecules (interleukin [IL] 1alpha, tumor necrosis factor [TNF]-alpha, and oxidized LDL [ox-LDL]) by use of Northern blot and ribonuclease protection assays for mRNA, Western blot and immunoprecipitation for protein, and gelatin zymography for catalytic activity. Medial SMCs of normal vessel wall expressed MT1-MMP. In atherosclerotic arteries, MT1-MMP expression was noted within the complex atheroma colocalizing with SMCs and macrophages (Mphi). Cultured SMCs constitutively expressed MT1-MMP mRNA and protein, which increased 2- to 4-fold over control in a time-dependent manner within 4 to 8 hours of exposure to IL-1alpha, TNF-alpha, and ox-LDL (thiobarbituric acid-reactive substances, 13.4 nmol/mg LDL protein), whereas native LDL had no effect. Flow cytometry revealed MT1-MMP expression by human monocyte-derived Mphi, which increased 3.8-fold over baseline within 6 hours after exposure to 10 ng/mL TNF-alpha. CONCLUSIONS: This study demonstrates that MT1-MMP, an activator of pro-MMP-2, is expressed by SMCs and Mphi in human atherosclerotic plaques. Furthermore, proinflammatory molecules upregulate MT1 MMP expression in vascular SMCs and Mphi. Thus, activation of SMCs and Mphi by proinflammatory molecules may influence extracellular matrix remodeling in atherosclerosis by regulating MT1-MMP expression. PMID- 10377073 TI - Expression of lectinlike oxidized low-density lipoprotein receptor-1 in human atherosclerotic lesions. AB - BACKGROUND: Oxidized LDL (Ox-LDL) seems to play key roles in atherogenesis. Lectinlike Ox-LDL receptor-1 (LOX-1) is a recently identified cell-surface receptor for Ox-LDL. The relationship of this novel receptor for Ox-LDL to atherogenesis, however, has not yet been clarified. In this study, we explored the expression of LOX-1 in the atherosclerotic lesions of human carotid arteries. METHODS AND RESULTS: Using carotid endarterectomy specimens obtained from 21 patients and 2 samples of normal human aortas, we examined LOX-1 expression by reverse transcription-polymerase chain reaction and immunohistochemistry. In aortas without atherosclerosis, LOX-1 expression was undetectable by immunohistochemistry and negligible by reverse transcription-polymerase chain reaction. In carotid arteries, luminal endothelial cells covering early atherosclerotic lesions were more frequently positive for LOX-1 expression than those in advanced atherosclerotic lesions. Endothelial cells in the intimal neovasculature of advanced lesions also expressed LOX-1. In addition, macrophages and smooth muscle cells in the intima of advanced atherosclerotic plaques were positive for LOX-1 expression. CONCLUSIONS: LOX-1 may play important roles in Ox LDL uptake and subsequent functional alteration in the luminal endothelium in early atherosclerotic lesions and in intimal neovascular endothelial cells in advanced plaques. Furthermore, LOX-1 may also be involved in Ox-LDL uptake and subsequent foam cell transformation in macrophages and smooth muscle cells in the atherosclerotic intima. PMID- 10377074 TI - Iron-dependent human platelet activation and hydroxyl radical formation: involvement of protein kinase C. AB - BACKGROUND: Iron is an important modulator of lipid peroxidation, and its levels have been associated with the progression of atherosclerosis. Little is known about the possibility that this metal, when released from tissue stores, may modulate the reactivity of blood cell components, in particular platelets. Therefore, we investigated a possible link between iron, oxygen free radical formation, and platelet function. METHODS AND RESULTS: Human whole blood was stimulated with collagen 2 micrograms/mL, and an irreversible aggregation with thromboxane (Tx)B2 formation was observed (15+/-4 versus 130+/-10 ng/mL). Deferoxamine (DSF), a specific iron chelator, and catalase, an H2O2 scavenger, inhibited collagen-induced whole-blood aggregation. The aggregation was accompanied by an increase in hydroxyl radical (OH.) levels (30+/-8 versus 205+/ 20 nmol/L dihydroxybenzoates), which were reduced by DSF and by 2 specific OH. scavengers, mannitol and deoxyribose. Iron (Fe2+) dose-dependently induced platelet aggregation, TxB2 formation (6+/-2 versus 135+/-8 ng/mL), and protein kinase C (PKC) translocation from the cytosol to the cell membrane when added to platelets that have been primed with a low concentration of collagen (0.2 micrograms/mL). In the same system, an increase in OH. levels was observed (37+/ 12 versus 230+/-20 nmol/L dihydroxybenzoates). Mannitol and deoxyribose, but not urea, were able to reduce OH. formation, PKC activation, and platelet aggregation. Selective inhibition of PKC activity by GF 109203X prevented iron dependent platelet aggregation without influencing OH. production. CONCLUSIONS: The present study shows that iron can directly interact with human platelets, resulting in their activation. Its action is mediated by OH. formation and involves PKC activity. Our findings provide an additional contribution to the understanding of the mechanism(s) by which iron overload might promote atherosclerosis and coronary artery disease. PMID- 10377075 TI - PPARalpha activators inhibit cytokine-induced vascular cell adhesion molecule-1 expression in human endothelial cells. AB - BACKGROUND: Adhesion molecule expression on the endothelial cell (EC) surface is critical for leukocyte recruitment to atherosclerotic lesions. Better understanding of transcriptional regulation of adhesion molecules in ECs may provide important insight into plaque formation. Peroxisome proliferator activated receptor-alpha (PPARalpha), a member of the nuclear receptor family, regulates gene expression in response to certain fatty acids and fibric acid derivatives. The present study investigated PPARalpha expression in human ECs and their regulation of vascular cell adhesion molecule-1 (VCAM-1). METHODS AND RESULTS: Immunohistochemistry revealed that human carotid artery ECs express PPARalpha. Pretreatment of cultured human ECs with the PPARalpha activators fenofibrate or WY14643 inhibited TNF-alpha-induced VCAM-1 in a time- and concentration-dependent manner, an effect not seen with PPARgamma activators. Both PPARalpha activators decreased cytokine-induced VCAM-1 mRNA expression without altering its mRNA half-life. Transient transfection of deletional VCAM-1 promoter constructs and electrophoretic mobility shift assays suggest that fenofibrate inhibits VCAM-1 transcription in part by inhibiting NF-kappaB. Finally, PPARalpha activators significantly reduced adhesion of U937 cells to cultured human ECs. CONCLUSIONS: Human ECs express PPARalpha, a potentially important regulator of atherogenesis through its transcriptional control of VCAM 1 gene expression. Such findings also have implications regarding the clinical use of lipid-lowering agents, like fibric acids, which can activate PPARalpha. PMID- 10377076 TI - Human coronary arteriolar dilation to bradykinin depends on membrane hyperpolarization: contribution of nitric oxide and Ca2+-activated K+ channels. AB - BACKGROUND: K+ channel activation in vascular smooth muscle cells (VSMCs) plays a key role in regulating vascular tone. It has been proposed that endothelium derived hyperpolarizing factor (EDHF) contributes to microvascular dilation more than nitric oxide (NO) does. Whether hyperpolarization is important for coronary arteriolar dilation in humans is not known. Bradykinin (BK), an endogenous vasoactive substance, is released from ischemic myocardium and regulates coronary resistance. Therefore, we tested the effects of inhibiting NO synthase, cyclooxygenase, and K+ channels on the changes in diameter and membrane potential (Em) in response to BK in isolated human coronary microvessels. METHODS AND RESULTS: Arterioles (97+/-4 micrometers; n=120) dissected from human right atrial appendages (n=78) were cannulated at a distending pressure of 60 mm Hg and zero flow. Changes in vessel diameter (video microscopy) and VSMC Em (glass microelectrodes) were measured simultaneously. In vessels constricted and depolarized (Em; -50+/-3 to -28+/-2 mV) with endothelin-1 (ET), dilation to BK was associated with greater membrane hyperpolarization (-48+/-3 mV at 10(-6) mol/L) than dilation to sodium nitroprusside (SNP) (-34+/-2 mV at 10(-4) mol/L) for similar degrees of dilation. Treatment with Nomega-nitro-L-arginine methyl ester (L-NAME; 10(-4) mol/L), an NO synthase inhibitor, partially decreased dilation to BK (maximum dilation 61+/-10% versus control 92+/-4%; P<0.05). Charybdotoxin (CTX; 10(-8) mol/L), a large-conductance Ca2+-activated K+ channel blocker, or apamin (10(-7) mol/L), a small-conductance Ca2+-activated K+ channel blocker, inhibited both dilation (CTX 22+/-6% and apamin 45+/-10% versus control 69+/-6%; P<0.05) and membrane hyperpolarization (CTX -31+/-2 mV and apamin -37+/ 2 mV versus control -44+/-2 mV; P<0.05) to BK, whereas glibenclamide (10(-6) mol/L), an ATP-sensitive K+ channel blocker, was without effect. CONCLUSIONS: Vasodilation of human coronary arterioles to BK is largely dependent on membrane hyperpolarization by Ca2+-activated K+ channel activation, with apparently less of a role for endothelium-derived NO. This suggests a role for K+ channel activation in regulating human coronary arteriolar tone. PMID- 10377077 TI - Improved coronary artery definition with T2-weighted, free-breathing, three dimensional coronary MRA. AB - BACKGROUND: Three-dimensional (3D) navigator-gated and prospectively corrected free-breathing coronary magnetic resonance angiography (MRA) allows for submillimeter image resolution but suffers from poor contrast between coronary blood and myocardium. Data collected over >100 ms/heart beat are also susceptible to bulk cardiac and respiratory motion. To address these problems, we examined the effect of a T2 preparation prepulse (T2prep) for myocardial suppression and a shortened acquisition window on coronary definition. METHODS AND RESULTS: Eight healthy adult subjects and 5 patients with confirmed coronary artery disease (CAD) underwent free-breathing 3D MRA with and without T2prep and with 120- and 60-ms data-acquisition windows. The T2prep resulted in a 123% (P<0. 001) increase in contrast-to-noise ratio (CNR). Coronary edge definition was improved by 33% (P<0.001). Acquisition window shortening from 120 to 60 ms resulted in better vessel definition (11%; P<0.001). Among patients with CAD, there was a good correspondence with disease. CONCLUSIONS: Free-breathing, T2prep, 3D coronary MRA with a shorter acquisition window resulted in improved CNR and better coronary artery definition, allowing the assessment of coronary disease. This approach offers the potential for free-breathing, noninvasive assessment of the major coronary arteries. PMID- 10377079 TI - Health-related quality of life after angioplasty and stent placement in patients with iliac artery occlusive disease: results of a randomized controlled clinical trial. The Dutch Iliac Stent Trial Study Group. AB - BACKGROUND: To assess the quality of life in patients with iliac artery occlusive disease, we compared primary stent placement versus primary angioplasty followed by selective stent placement in a multicenter randomized controlled trial. METHODS AND RESULTS: Quality-of-life assessments were completed by 254 patients in a telephone interview. Assessment measures consisted of the RAND 36-Item Health Survey 1.0, time tradeoff, standard gamble, rating scale, health utilities index, and EuroQol-5D. The interviews were performed before treatment and after 1, 3, 12, and 24 months. When the 2 treatments were compared, no significant difference was observed (P>0.05). All measurements showed a significant improvement in the quality of life after treatment (P<0.05). The RAND 36-Item Health Survey measures physical functioning, role limitations caused by physical problems, and bodily pain and the EuroQol-5D were the most sensitive to the impact of revascularization. CONCLUSIONS: Health-related quality of life improves equally after primary stent placement and primary angioplasty with selective stent placement in the treatment of intermittent claudication caused by iliac artery occlusive disease. PMID- 10377078 TI - Preintervention arterial remodeling as an independent predictor of target-lesion revascularization after nonstent coronary intervention: an analysis of 777 lesions with intravascular ultrasound imaging. AB - BACKGROUND: Pathological and intravascular ultrasound (IVUS) studies have documented arterial remodeling during atherogenesis. However, the impact of this remodeling process on the long-term outcome after percutaneous intervention is unknown. METHODS AND RESULTS: We used preintervention IVUS to define positive and negative/intermediate remodeling in a total of 777 lesions in 715 patients treated with nonstent techniques. Positive remodeling (lesion external elastic membrane area greater than average reference) was present in 313 lesions; intermediate/negative remodeling (lesion external elastic membrane area less than or equal to reference) was present in the other 464. Baseline clinical and angiographic characteristics were similar, except for a slightly higher percentage of insulin-dependent diabetic patients (10.2% versus 6.1%; P=0.054) in the negative/intermediate-remodeling group. Angiographic success and in-hospital and short-term complications were comparable in the 2 groups. There was no significant correlation between remodeling (as a continuous variable) and final lumen area (r=0.06) or final lesion plaque burden (r=0.17). At 18+/-13 months of clinical follow-up, both groups had similar rates of death and Q-wave myocardial infarction: 3.4% and 2.5% for the negative/intermediate-remodeling group versus 2.7% and 2.7% for the positive-remodeling group. However, the target-lesion revascularization (TLR) rate was 20.2% for the negative/intermediate-remodeling group versus 31.2% for the positive-remodeling group (P=0.007), and remodeling, as a continuous variable, was strongly correlated with probability of TLR (P=0.0001). By multivariable logistic regression analysis, diabetes (OR=2.3), left anterior descending artery location (OR=1.8), and remodeling (OR=5.9) were independent predictors of TLR. CONCLUSIONS: Positive lesion-site remodeling is associated with a higher long-term TLR after a nonstent interventional procedure. Thus, long-term clinical outcome appears to be determined in part by preintervention lesion characteristics. PMID- 10377080 TI - QT interval is linked to 2 long-QT syndrome loci in normal subjects. AB - BACKGROUND: The rate-corrected QT interval (QTc) is heritable, and the discovery of quantitative trait loci that influence the QTc would be an important step in identifying the genes responsible for life-threatening arrhythmias in the general population. We studied 66 pairs of unselected normal dizygotic (DZ) twin subjects and their parents in a sib-pair analysis. We tested for linkage of gene loci harboring genes known to cause the long-QT syndrome (LQT) to the quantitative trait QTc. METHODS AND RESULTS: We found genetic variance on QRS duration, QRS axis, T-wave axis, and QTc. Women had a longer QTc than men. Microsatellite markers were tested in the vicinity of the gene loci for the 5 known LQT genes. We found significant linkage of QTc with the loci for LQT1 on chromosome 11 and LQT4 on chromosome 4 but not to LQT2, LQT3, or LQT5. We also found linkage of the QRS axis with LQT2 and LQT3. CONCLUSIONS: We suggest that these quantitative trait loci may represent the presence of variations in LQT genes that could be important to the risk for rhythm disturbances in the general population. PMID- 10377081 TI - Congenital long-QT syndrome caused by a novel mutation in a conserved acidic domain of the cardiac Na+ channel. AB - BACKGROUND: Congenital long-QT syndrome (LQTS) is an inherited condition of abnormal cardiac excitability characterized clinically by an increased risk of ventricular tachyarrhythmias. One form, LQT3, is caused by mutations in the cardiac voltage-dependent sodium channel gene, SCN5A. Only 5 SCN5A mutations have been associated with LQTS, and more work is needed to improve correlations between SCN5A genotypes and associated clinical syndromes. METHODS AND RESULTS: We researched a 3-generation white family with autosomal dominant LQTS who exhibited a wide clinical spectrum from mild bradycardia to sudden death. Molecular genetic studies revealed a single nucleotide substitution in SCN5A exon 28 that caused the substitution of Glu1784 by Lys (E1784K). The mutation occurs in a highly conserved domain within the C-terminus of the cardiac sodium channel containing multiple, negatively charged amino acids. Two-electrode voltage-clamp recordings of a recombinant E1784K mutant channel expressed in Xenopus oocytes revealed a defect in fast inactivation characterized by a small, persistent current during long membrane depolarizations. Coexpression of the mutant with the human sodium channel beta1-subunit did not affect the persistent current, even though we did observe shifts in the voltage dependence of steady-state inactivation. Neutralizing multiple, negatively charged residues in the same region of the sodium channel C-terminus did not cause a more severe functional defect. CONCLUSIONS: We characterized the genetics and molecular pathophysiology of a novel SCN5A sodium channel mutation, E1784K. The functional defect exhibited by the mutant channel causes delayed myocardial repolarization, and our data on the effects of multiple charge neutralizations in this region of the C-terminus suggest that the molecular mechanism of channel dysfunction involves an allosteric rather than a direct effect on channel gating. PMID- 10377082 TI - Familial hypertrophic cardiomyopathy in maine coon cats: an animal model of human disease. AB - BACKGROUND: A naturally occurring animal model of familial hypertrophic cardiomyopathy (FHCM) is lacking. We identified a family of Maine coon cats with HCM and developed a colony to determine mode of inheritance, phenotypic expression, and natural history of the disease. METHODS AND RESULTS: A proband was identified, and related cats were bred to produce a colony. Affected and unaffected cats were bred to determine the mode of inheritance. Echocardiography was used to identify affected offspring and determine phenotypic expression. Echocardiograms were repeated serially to determine the natural history of the disease. Of 22 offspring from breeding affected to unaffected cats, 12 (55%) were affected. When affected cats were bred to affected cats, 4 (45%) of the 9 were affected, 2 (22%) unaffected, and 3 (33%) stillborn. Findings were consistent with an autosomal dominant mode of inheritance with 100% penetrance, with the stillborns representing lethal homozygotes that died in utero. Affected cats usually did not have phenotypic evidence of HCM before 6 months of age, developed HCM during adolescence, and developed severe HCM during young adulthood. Papillary muscle hypertrophy that produced midcavitary obstruction and systolic anterior motion of the mitral valve was the most consistent manifestation of HCM. Cats died suddenly (n=5) or of heart failure (n=3). Histopathology of the myocardium revealed myocardial fiber disarray, intramural coronary arteriosclerosis, and interstitial fibrosis. CONCLUSIONS: HCM in this family of Maine coon cats closely resembles the human form of FHCM and should prove a valuable tool for studying the gross, cellular, and molecular pathophysiology of the disease. PMID- 10377083 TI - Evidence for a causal role of the renin-angiotensin system in nitrate tolerance. AB - BACKGROUND: We have previously shown that nitroglycerin (NTG) therapy increases vascular expression of endothelin 1 (ET-1) and stimulates vascular superoxide (O2.-) production via activation of NADH/NADPH oxidases. Both phenomena are stimulated by angiotensin II in vitro, and the renin-angiotensin system is activated during early nitrate therapy. We hypothesized that either angiotensin II or ET-1 may increase vascular O2.- production during nitrate therapy. METHODS AND RESULTS: In New Zealand White rabbits, 3 days of treatment with NTG patches increased plasma renin activity for the entire treatment period. After 24 hours of NTG treatment, angiotensin II type 1 (AT1) receptor expression and vascular ACE activity were significantly decreased. At this time, constrictions to angiotensin I and II were depressed, but there was no loss of NTG vasodilator potency. Within 3 days of continuous NTG treatment, relaxations to NTG were markedly blunted. This was associated with an increase in AT1 receptor mRNA expression, a return of ACE activity back to baseline, and a marked increase in constrictions to angiotensin I and II despite continuously increased plasma renin activity. Tolerance was associated with a 2-fold increase in vascular O2.-, as estimated by lucigenin-enhanced chemiluminescence. Concomitant treatment with the AT1 receptor antagonist losartan (5 to 25 mg. kg-1. d-1) dose-dependently normalized vascular O2.- and prevented tolerance to NTG and cross-tolerance to endogenous nitric oxide released by acetylcholine. The nonselective ET-1 receptor blocker bosentan (100 mg. kg-1. d-1) had similar but less pronounced effects. CONCLUSIONS: The positive effects of AT1 and ET-1 receptor blockade on tolerance and O2.- production imply a pathophysiological role for angiotensin II and to some extent for ET-1 in the development of nitrate tolerance. PMID- 10377084 TI - Impaired collateral vessel development associated with reduced expression of vascular endothelial growth factor in ApoE-/- mice. AB - BACKGROUND: The impact of disordered lipid metabolism on collateral vessel development was studied in apolipoprotein (apo)E-/- mice with unilateral hindlimb ischemia. METHODS AND RESULTS: Hindlimb blood flow and capillary density were markedly reduced in apoE-/- mice versus C57 controls. This was associated with reduced expression of vascular endothelial growth factor (VEGF) in the ischemic limbs of apoE-/- mice. Cell-specific immunostaining localized VEGF protein expression to skeletal myocytes and infiltrating T cells in the ischemic limbs of C57 mice; in contrast, T-cell infiltrates in ischemic limbs of apoE-/- mice were severely reduced. The critical contribution of T cells to VEGF expression and collateral vessel growth was reinforced by the finding of accelerated limb necrosis in athymic nude mice with operatively induced hindlimb ischemia. Adenoviral VEGF gene transfer to apoE-/- mice resulted in marked augmentation of hindlimb blood flow and capillary density. CONCLUSIONS: These findings thus underscore the extent to which hyperlipidemia adversely affects native collateral development but does not preclude augmented collateral vessel growth in response to exogenous cytokines. Moreover, results obtained in the apoE-/- and athymic nude mice imply a critical role for infiltrating T cells as a source of VEGF in neovascularization of ischemic tissues. PMID- 10377086 TI - Spot welding the gap in atrial flutter ablation. PMID- 10377085 TI - Expression of tissue inhibitor of matrix metalloproteinases 1 by use of an adenoviral vector inhibits smooth muscle cell migration and reduces neointimal hyperplasia in the rat model of vascular balloon injury. AB - BACKGROUND: Cell migration is a major contributor to injury-induced neointimal hyperplasia and depends on alteration of the proteolytic balance within the arterial wall toward matrix breakdown. This is partly mediated by the matrix metalloproteinases (MMPs) and their natural inhibitors, the tissue inhibitors of metalloproteinases (TIMPs). METHODS AND RESULTS: An increase in expression of biologically active and immunoreactive TIMP-1 was seen in vitro after infection of rat smooth muscle cells (SMCs) with Av1.TIMP1 (an adenoviral vector containing the human TIMP1 cDNA). Infection of rat SMCs with Av1.TIMP1 reduced migration in vitro by 27% compared with control virus-infected cells (37.6+/-4.34 versus 51+/ 5.01 cells per high-power field, P<0.05). The adenoviral vector was delivered to the injured rat carotid artery, and 4 days later, immunoreactive protein was identified and migration of SMCs reduced by 60% (5.2+/-0. 5 versus 12.8+/-1.5 cells per section, P<0.05, n=5). Neointimal area 14 days after injury showed a 30% reduction in the animals receiving the Av1.TIMP1 virus compared with controls (0.09+/-0.01 versus 0. 14+/-0.01 mm2, P=0.02, n=14). CONCLUSIONS: The response to arterial balloon injury involves MMP-dependent SMC migration and can be attenuated in vivo by the transmural expression of TIMP-1 by adenoviral gene transfer. PMID- 10377087 TI - Electron beam computed tomographic angiography and 3-dimensional reconstruction of a stented saphenous vein graft. PMID- 10377088 TI - Effects of environmental pH on membrane proteins in Borrelia burgdorferi. AB - Borrelia burgdorferi, the causative agent of Lyme disease, alternates between the microenvironments of the tick vector, Ixodes scapularis, and a mammalian host. The environmental conditions the spirochete encounters during its infectious cycle are suspected to differ greatly in many aspects, including available nutrients, temperature, and pH. Here we identify alterations in the membrane protein profile, as determined by immunoblotting and two-dimensional nonequilibrium pH gradient gel electrophoresis (2D-NEPHGE), that occur in virulent B. burgdorferi B31 as the pH of the medium is altered. Initial comparisons of cultures incubated at pHs 6.0, 7.0, and 8.0 yielded alterations in the expression of seven membrane proteins as determined by probing with hyperimmune rabbit serum. Six of these membrane proteins (54, 45, 44, 43, 35, and 24 kDa) were either present in increased amounts in or solely expressed by cultures incubated at pHs 6.0 and 7.0. The 24-kDa protein that decreased in expression at pH 8.0 was identified as outer surface protein C (OspC). In addition, a 42-kDa membrane protein increased in amount in cultures incubated at pH 8.0. Similar changes were observed with serum from a mouse infected by tick bite, with the recognition of two additional bands (48 and 46 kDa) unique to pHs 6. 0 and 7.0. When membrane fractions were analyzed by 2D-NEPHGE, at least 37 changes in the membrane protein profile between cells incubated at pHs 6.0, 7.0, and 8.0 were observed by immunoblotting and silver staining. Environmental cues such as pH may prove important in the regulation of virulence determinants and factors necessary for the adaptation of B. burgdorferi to the tick or mammalian microcosm. PMID- 10377089 TI - Effects of Mycoplasma fermentans incognitus on differentiation of THP-1 cells. AB - Mycoplasma fermentans incognitus has been isolated from human tissue in patients both with and without AIDS who died of systemic infection. M. fermentans incognitus and other strains of M. fermentans have been associated with rheumatoid arthritis. While cell extracts of M. fermentans incognitus can induce changes in murine and human cells of the monocytic lineage, little is known about interactions of viable organisms with such cells. Because of the central role of macrophages in chronic inflammation, we examined the effects of M. fermentans incognitus on surface markers and functions of THP-1 cells, a well-characterized human monocytic cell line. This cell line has been used extensively in studies of macrophage differentiation, especially following exposure to phorbol esters. Changes in cell morphology, phagocytosis, rate of cell division, and selected surface markers were evaluated in cultures of THP-1 cells exposed to phorbol myristate acetate (PMA), M. fermentans incognitus, or both. As reported by other investigators, PMA induced THP-1 cells to differentiate into cells resembling tissue macrophages. M. fermentans incognitus only minimally affected changes induced by PMA, slightly increasing the percentage of cells positive for FCgammaRI and major histocompatibility complex (MHC) class II antigens. M. fermentans incognitus alone induced an incomplete arrest in the cell cycle at G0 phase, increased phagocytic ability, and enhanced expression of FCgammaRI, CR3, CR4, and MHC class II antigens. PMID- 10377090 TI - Unanticipated heterogeneity in growth rate and virulence among Candida albicans AAF1 null mutants. AB - The disruption of a specific gene in Candida albicans is commonly used to determine the function of the gene product. We disrupted AAF1, a gene of C. albicans that causes Saccharomyces cerevisiae to flocculate and adhere to endothelial cells. We then characterized multiple heterozygous and homozygous mutants. These null mutants adhered to endothelial cells to the same extent as did the parent organism. However, mutants with presumably the same genotype revealed significant heterogeneity in their growth rates in vitro. This heterogeneity was not the result of the transformation procedure per se, nor was it caused by differences in the expression or function of URA3, a marker used in the process of gene disruption. The growth rate among the different heterozygous and homozygous null mutants was positively correlated with in vivo virulence in mice. It is possible that the variable phenotypes of C. albicans were due to mutations outside of the AAF1 coding region that were introduced during the gene disruption process. These results indicate that careful phenotypic characterization of mutants of C. albicans generated through targeted gene disruption should be performed to exclude the introduction of unexpected mutations that may influence pathogenicity in mice. PMID- 10377091 TI - Survival of Mycobacterium avium and Mycobacterium tuberculosis in acidified vacuoles of murine macrophages. AB - Despite the antimicrobial mechanisms of vertebrate phagocytes, mycobacteria can survive within the phagosomes of these cells. These organisms use various strategies to evade destruction, including inhibition of acidification of the phagosome and inhibition of phagosome-lysosome fusion. In contrast to mycobacteria, Coxiella burnetii, the etiologic agent of Q fever, inhabits a spacious acidified intracellular vacuole which is prone to fusion with other vacuoles of the host cell, including phagosomes containing mycobacteria. The Coxiella-infected cell thus provides a unique model for investigating the survival of mycobacteria in an acidified phagosome-like compartment. In the present study, murine bone marrow-derived macrophages were infected with either Mycobacterium avium or Mycobacterium tuberculosis and then coinfected with C. burnetii. We observed that the majority of phagocytosed mycobacteria colocalized to the C. burnetii-containing vacuole, which maintained its acidic properties. In coinfected macrophages, the growth of M. avium was not impaired following fusion with the acidified vacuole. In contrast, the growth rate of M. tuberculosis was reduced in acidified vacuoles. These results suggest that although both species of mycobacteria inhibit phagosome-lysosome fusion, they may be differentially susceptible to the toxic effects of the acidic environment in the mature phagolysosome. PMID- 10377092 TI - Pili binding to asialo-GM1 on epithelial cells can mediate cytotoxicity or bacterial internalization by Pseudomonas aeruginosa. AB - The interaction of Pseudomonas aeruginosa type IV pili and the glycosphingolipid asialo-GM1 (aGM1) can mediate bacterial adherence to epithelial cells, but the steps subsequent to this adherence have not been elucidated. To investigate the result of the interaction of pili and aGM1, we used polarized epithelial monolayers of Madin-Darby canine kidney (MDCK) cells in culture, which contained little detectable aGM1 on their apical surface but were able to incorporate exogenous aGM1. Compared to an untreated monolayer, P. aeruginosa PA103 displayed an eightfold increase in association with and fivefold more cytotoxicity toward MDCK cells pretreated with aGM1. Cytotoxicity of either carrier-treated or aGM1 treated monolayers required the type III secreted protein ExoU. Asialo-GM1 pretreatment of MDCK monolayers likewise augmented bacterial internalization of an isogenic invasive strain approximately fourfold. These increases were not seen in monolayers treated with GM1, the sialyated form of the glycolipid, and were inhibited by treatment with an antibody to aGM1. Also, the aGM1-mediated adhesion, cytotoxicity, and internalization required intact type IV pili since nonpiliated PA103 mutants were unaffected by aGM1 pretreatment of MDCK cells. These results demonstrate that epithelial cell injury and bacterial internalization can proceed from the same adhesin-receptor interaction, and they indicate that P. aeruginosa exoproducts solely determine the steps subsequent to adhesion. PMID- 10377093 TI - Moesin functions as a lipopolysaccharide receptor on human monocytes. AB - Bacterial endotoxin (lipopolysaccharide [LPS]), a glycolipid found in the outer membranes of gram-negative bacteria, induces the secretion of proinflammatory cytokines such as tumor necrosis factor alpha (TNF-alpha), interleukin-1 (IL-1), and IL-6 by monocytes/macrophages. The secretion of these biologically active compounds leads to multiple pathological conditions, such as septic shock. There is substantial evidence that chronic exposure to LPS mediates, at least in part, the tissue destruction associated with gram-negative infection. CD14, a 55-kDa protein, has been identified as an LPS receptor. In conjunction with a serum protein, LPS binding protein (LBP), LPS-CD14 interactions mediate many LPS functions in the inflammatory response. However, CD14 lacks a cytoplasmic domain, or any known signal transduction sequence motif, suggesting the existence of another cell surface domain capable of transducing signals. In this paper, we report a second, CD14-independent LPS binding site, which, based on biological activity, appears to be a functional LPS receptor. Cross-linking experiments were performed to identify LPS binding sites. Two molecules were identified: a 55-kDa protein (CD14) and a second, 78-kDa band. Sequencing of the 78-kDa protein by mass spectroscopic analysis revealed 100% homology with moesin (membrane organizing extension spike protein). Antibody to CD14 induced partial blocking of the LPS response. However, antimoesin monoclonal antibody completely blocked the LPS-induced TNF-alpha response in human monocytes, without blocking CD14 binding of LPS. Irrelevant isotype controls had no effect. Additional experiments were performed to evaluate the specificity of the antimoesin blocking. Separate experiments evaluated antimoesin effects on monocyte chemotaxis, IL-1 production in response to IL-1 stimulation, and TNF-alpha secretion in response to Staphylococcus aureus stimulation. Antimoesin blocked only LPS-mediated events. The data suggest that moesin functions as an independent LPS receptor on human monocytes. The role of moesin in transduction of CD14-mediated signals is discussed. PMID- 10377094 TI - T-Cell hyporesponsiveness induced by activated macrophages through nitric oxide production in mice infected with Mycobacterium tuberculosis. AB - In active tuberculosis, T-cell response to Mycobacterium tuberculosis is known to be reduced. In the course of Mycobacterium tuberculosis infection in mice, we observed that T-cell proliferation in response to M. tuberculosis purified protein derivative (PPD) reached the maximum level on day 7, then declined to the minimal level on day 14, and persisted at a low level through day 28 postinfection. The frequency of PPD-specific CD4 T cells in the spleen on day 28 decreased to one-sixth on day 7. To further investigate the mechanism of this T cell hyporesponsiveness, we next analyzed the suppressive activity of spleen macrophages on T-cell function. The nonspecific proliferative response of naive T cells and the PPD-specific proliferative response of T cells were suppressed by day 28 macrophages, but not by day 7 macrophages or naive macrophages. This reduction of proliferative response was restored by addition of nitric oxide synthesis inhibitor, NG-monoethyl-L-arginine monoacetate, but not by monoclonal antibody against interleukin 10 or transforming growth factor beta. These data indicate that the macrophages from mice chronically infected with M. tuberculosis suppress T-cell response through production of nitric oxide, suggesting that nitric oxide-induced elimination mediated by activated macrophages may reduce the T-cell response and the number of mycobacterium-specific CD4 T cells in vivo. PMID- 10377095 TI - Promoter architecture of the Porphyromonas gingivalis fimbrillin gene. AB - Porphyromonas gingivalis fimbriae can mediate adherence to many of the available substrates in the oral cavity. Expression of P. gingivalis fimbriae is regulated at the transcriptional level by environmental signals, such as temperature and hemin concentration. The arrangement of the upstream promoter and regulatory sequences required for transcription and control of the fimbrial structural gene (fimA) was investigated. Primer extension analysis demonstrated that the transcriptional start site of the fimA gene is located 41 bp upstream from the translational start codon. A region (upf) spanning 648 bp upstream of the start codon to 44 bp downstream of the translational start site was cloned upstream of a promoterless lacZ reporter gene. A series of deletion and base substitution mutations were then generated in the upf region. The constructs were introduced into the chromosome of P. gingivalis, and promoter activity measured by assaying levels of beta-galactosidase. The results showed that fimA contains sequences resembling sigma70 promoter consensus sequences, consisting of a -10 region (TATGAC) located at -18 to -23 and a -35 region (TTGTTG) located at -41 to -46 from the transcriptional start point. The AT-rich upstream sequences spanning bases -48 to -85 and bases -90 to -240 were required for full expression of the fimA gene, indicating the existence of positive regulation regions. Moreover, the -48 to -64 region may constitute an UP element, contributing to promoter activity in P. gingivalis. Thus, our data suggest that the P. gingivalis fimA gene has a transcription complex consisting of -10 and -35 sequences, an UP element, and additional AT-rich upstream regulatory sequences. PMID- 10377096 TI - Coxiella burnetii infection increases transferrin receptors on J774A. 1 cells. AB - Inoculation with viable, but not inactivated, Coxiella burnetii resulted in the increased expression of transferrin receptors (TfR) in the murine macrophage-like cell line J774A.1. This upregulation was evident in immunoblots as early as 6 h postinfection, with TfR levels continuing to increase through the first 24 h of infection. Fluorescent labeling revealed that TfR upregulation occurred throughout infected monolayers, eliminating the possibility that it reflected a response by a minor subset of host cells. In addition, TfR trafficking did not appear to be affected by C. burnetii infection. Consistent with the increase in TfRs, inoculation with viable C. burnetii resulted in a 2.5-fold increase in total cellular iron by 12 h postinoculation. Our further findings that the chelation of intracellular iron arrests C. burnetii replication and that C. burnetii metabolic activities in vitro are affected by iron concentration suggest that TfR upregulation is a salient factor in C. burnetii infection, and we speculate that it may represent a significant virulence mechanism. PMID- 10377097 TI - Increase in gamma interferon-secreting CD8(+), as well as CD4(+), T cells in lungs following aerosol infection with Mycobacterium tuberculosis. AB - Although it is well established that CD4(+) T cells are required for the protective immune response against tuberculosis (TB), there is some evidence that CD8(+) T cells are also involved in the host response to Mycobacterium tuberculosis. There is, however, a paucity of information on the pulmonary CD8(+) T-cell response during infection. We therefore have compared the changes in both CD8(+) and CD4(+) T cells following aerosol infection with M. tuberculosis. There was an observed delay between the peak of infection and the activated T-cell response in the lung. The kinetics of CD8(+) and CD4(+) T-cell responses in the lung were identical, both peaking at week 8, 4 weeks later than the peak of cellular response in draining lymph nodes. Similar changes in activation/memory phenotypes occurred on the pulmonary CD8(+) and CD4(+) T cells. Following in vitro restimulation, both subsets synthesized gamma interferon, a cytokine essential for controlling M. tuberculosis infection. Since lung CD8(+) T cells are actively expanded during aerosol M. tuberculosis infection, it is important that both CD8(+) and CD4(+) T cells be targeted in the design of future TB vaccines. PMID- 10377098 TI - Purification, characterization, and sequence analysis of a potential virulence factor from Porphyromonas gingivalis, peptidylarginine deiminase. AB - The initiation and progression of adult-onset periodontitis has been associated with infection of the gingival sulcus by Porphyromonas gingivalis. This organism utilizes a multitude of virulence factors to evade host defenses as it establishes itself as one of the predominant pathogens in periodontal pockets. A feature common to many other oral pathogens is the production of ammonia due to its protective effect during acidic cleansing cycles in the mouth. Additionally, ammonia production by P. gingivalis has been proposed as a virulence factor due to its negative effects on neutrophil function. In this study, we describe the first purification of a peptidylarginine deiminase (PAD) from a prokaryote. PAD exhibits biochemical characteristics and properties that suggest that it may be a virulence agent. PAD deiminates the guanidino group of carboxyl-terminal arginine residues on a variety of peptides, including the vasoregulatory peptide-hormone bradykinin, to yield ammonia and a citrulline residue. The soluble protein has an apparent mass of 46 kDa, while the DNA sequence predicts a full-length protein of 61.7 kDa. PAD is optimally active at 55 degrees C, stable at low pH, and shows the greatest activity above pH 9.0. Interestingly, in the presence of stabilizing factors, PAD is resistant to limited proteolysis and retains significant activity after short-term boiling. We propose that PAD, acting in concert with arginine specific proteinases from P. gingivalis, promotes the growth of the pathogen in the periodontal pocket, initially by enhancing its survivability and then by assisting the organism in its circumvention of host humoral defenses. PMID- 10377099 TI - Comparative analysis and immunological characterization of the Borrelia Bdr protein family. AB - Multiple circular and linear plasmids of Lyme disease and relapsing fever Borrelia spirochetes carry genes for members of the Bdr (Borrelia direct repeat) protein family. To define their common and divergent attributes, we first comprehensively compared the known homologs. Bdr proteins with predicted sizes ranging from 10.7 to 30. 6 kDa formed five homology groups, based on variable numbers of short direct repeats in a central domain and diverse N- and C-terminal domains. In a further characterization, Western blots were probed with rabbit antisera raised against either of two purified recombinant Bdr proteins from Borrelia burgdorferi B31. The results showed that antibodies cross-react and several Bdr paralogs 19.5 to 30.5 kDa in size are expressed by cultured strain B31 in a temperature-independent manner. In situ proteolysis, immunofluorescence, and growth inhibition assays indicated that Bdr proteins are not surface exposed. Distinct patterns of cross-reacting proteins of 17.5 to 33 kDa were also detected in other B. burgdorferi, Borrelia garinii, and Borrelia afzelii strains as well as in relapsing fever spirochetes Borrelia hermsii and Borrelia turicatae. Last, we examined whether these proteins are antibody targets during Lyme disease. Analysis of 47 Lyme disease patient sera by immunoblotting and enzyme-linked immunosorbent assays showed that 24 (51%) and 20 (43%), respectively, had detectable antibodies to one or more of the Bdr proteins. Together, these data indicate that Bdr proteins constitute a family of cross-reactive Borrelia proteins which are expressed in the course of Lyme disease and in vitro. PMID- 10377100 TI - Innate antimicrobial activity of nasal secretions. AB - Minimally manipulated nasal secretions, an accessible form of airway surface fluid, were tested against indigenous and added bacteria by using CFU assays. Antimicrobial activity was found to vary between donors and with different target bacteria and was markedly diminished by dilution of the airway secretions. Donor to-donor differences in electrophoresis patterns of nasal secretions in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (PAGE) and acid urea-PAGE analyses were readily observed, suggesting that polymorphic genes encode the secreted proteins. Three donors (of twenty-four total), whose nasal fluid yielded similar protein band patterns and did not kill indigenous bacteria, were determined to be heavy nasal carriers of Staphylococcus aureus. Their fluid was deficient in microbicidal activity toward a colonizing strain of S. aureus but the defect was corrected in vitro by a 1:1 addition of nasal fluid from noncarriers. The microbicidal activity of normal fluid was inactivated by heating it for 10 min to 100 degrees C and could not be restored solely by the addition of two major nasal antimicrobial proteins, lysozyme and lactoferrin. Several other known antimicrobial proteins and peptides, including statherin, secretory phospholipase A2, and defensins, were identified in nasal secretions and likely contribute to their total antimicrobial properties. Nasal fluid may serve as a useful model for the analysis of lower-airway secretions and their role in host defense against airway colonization and pulmonary infections. PMID- 10377101 TI - noxR3, a novel gene from Mycobacterium tuberculosis, protects Salmonella typhimurium from nitrosative and oxidative stress. AB - Reactive oxygen intermediates (ROI) and reactive nitrogen intermediates (RNI) produced by activated macrophages participate in host defense against the facultative intracellular pathogens Mycobacterium tuberculosis and Salmonella typhimurium. To survive within macrophages, such pathogens may have evolved ROI and RNI resistance mechanisms. ROI resistance pathways have been intensively studied. Much less is known about the mechanisms of resistance to RNI. To identify possible RNI resistance genes in M. tuberculosis, a mycobacterial library was expressed in S. typhimurium and subjected to selection by exposure to the NO donor S-nitrosoglutathione (GSNO) in concentrations sufficient to kill the vast majority of nontransformed salmonellae. Among the rare surviving recombinants was a clone expressing noxR3, a novel and previously anonymous M. tuberculosis gene predicted to encode a small, basic protein. Expression of noxR3 protected S. typhimurium not only from GSNO and acidified nitrite but also from H2O2. noxR3 is the third gene cloned from M. tuberculosis that has been shown to protect heterologous cells from both RNI and ROI. This suggests diversity in the repertoire of mechanisms that help pathogens resist the oxidative and nitrosative defenses of the host. PMID- 10377102 TI - Carrageenan primes leukocytes to enhance lipopolysaccharide-induced tumor necrosis factor alpha production. AB - We have previously reported that pretreatment with carrageenan (CAR) enhances lipopolysaccharide (LPS)-induced tumor necrosis factor alpha (TNF-alpha) production in and lethality for mice. Whole blood cultured in vitro was used to show that CAR pretreatment results in about a 200-fold increase in LPS-induced TNF-alpha production. CAR by itself did not induce TNF-alpha production. However, CAR-treated cultured medium sensitized whole blood to make more LPS-induced TNF than did saline-treated cultured medium in vitro. It was also demonstrated that CAR pretreatment increases TNF-alpha mRNA levels of both blood cells and peritoneal exudate cells, but not of bone marrow cells. Immunoelectron microscopic analysis revealed that polymorphonuclear leukocytes and macrophages are TNF-alpha-producing cells in CAR-treated mice. In CAR-treated mice, TNF-alpha was seen early after LPS injection in leukocytes in hepatic sinusoids and on the surfaces of endothelial cells. TNF-alpha was also detected late after LPS injection in hepatocytes which become edematous. These results suggest that CAR primes leukocytes to produce TNF-alpha in response to LPS and that they play an important role in the pathogenesis of liver injury. PMID- 10377103 TI - Cytotoxic T-lymphocyte epitopes fused to anthrax toxin induce protective antiviral immunity. AB - We have investigated the use of the protective antigen (PA) and lethal factor (LF) components of anthrax toxin as a system for in vivo delivery of cytotoxic T lymphocyte (CTL) epitopes. During intoxication, PA directs the translocation of LF into the cytoplasm of mammalian cells. Here we demonstrate that antiviral immunity can be induced in BALB/c mice immunized with PA plus a fusion protein containing the N-terminal 255 amino acids of LF (LFn) and an epitope from the nucleoprotein (NP) of lymphocytic choriomeningitis virus. We also demonstrate that BALB/c mice immunized with a single LFn fusion protein containing NP and listeriolysin O protein epitopes in tandem mount a CTL response against both pathogens. Furthermore, we show that NP-specific CTL are primed in both BALB/c and C57BL/6 mice when the mice are immunized with a single fusion containing two epitopes, one presented by Ld and one presented by Db. The data presented here demonstrate the versatility of the anthrax toxin delivery system and indicate that this system may be used as a general approach to vaccinate outbred populations against a variety of pathogens. PMID- 10377104 TI - Differences in the carboxy-terminal (Putative phospholipid binding) domains of Clostridium perfringens and Clostridium bifermentans phospholipases C influence the hemolytic and lethal properties of these enzymes. AB - The phospholipases C of C. perfringens (alpha-toxin) and C. bifermentans (Cbp) show >50% amino acid homology but differ in their hemolytic and toxic properties. We report here the purification and characterisation of alpha-toxin and Cbp. The phospholipase C activity of alpha-toxin and Cbp was similar when tested with phosphatidylcholine in egg yolk or in liposomes. However, the hemolytic activity of alpha-toxin was more than 100-fold that of Cbp. To investigate whether differences in the carboxy-terminal domains of these proteins were responsible for differences in the hemolytic and toxic properties, a hybrid protein (NbiCalpha) was constructed comprising the N domain of Cbp and the C domain of alpha-toxin. The hemolytic activity of NbiCalpha was 10-fold that of Cbp, and the hybrid enzyme was toxic. These results confirm that the C-terminal domain of these proteins confers different properties on the enzymatically active N terminal domain of these proteins. PMID- 10377105 TI - Role of lactosyl glycan sequences in inhibiting enteropathogenic Escherichia coli attachment. AB - Previously, we found that asialo-lactosamine sequences served as receptors for enteropathogenic Escherichia coli (EPEC) binding to Chinese hamster ovary (CHO) cells. In the present report, we have extended these earlier results by examining the ability of lactosamine- or fucosylated lactosamine-bovine serum albumin (BSA) glycoconjugates to inhibit EPEC, strain E2348/69, binding to HEp-2 cells. We found that, consistent with our previous findings with CHO cells, N acetyllactosamine-BSA was the most effective inhibitor of EPEC localized adherence to HEp-2 cells, with Lewis X-BSA being the next best inhibitor. Further investigation revealed that coincubating EPEC E2348/69 with these BSA glycoconjugates alone caused a decrease in the expression of the bundle-forming pilus structural subunit (BfpA) and intimin by the bacteria. BfpA and intimin expression were reduced to the greatest extent by N-acetyllactosamine-BSA and Lewis X-BSA, respectively. These results suggest that the glycoconjugate inhibition of EPEC binding to HEp-2 cells might be achieved, wholly or in part, by an active mechanism that is distinct from simple competitive antagonism of receptor-adhesin interactions. PMID- 10377106 TI - Green fluorescent protein as a marker in Rickettsia typhi transformation. AB - Transformation of rickettsiae is a recent accomplishment, but utility of this technique is limited due to the paucity of selectable markers suitable for use in this intracellular pathogen. We chose a green fluorescent protein variant optimized for fluorescence under UV lights (GFPUV) as a fluorometric marker and transformed Rickettsia typhi with an rpoB-GFPUV fusion construct. The rickettsiae were subsequently grown in Vero cells, and cultures were screened by PCR and restriction fragment length polymorphism (RFLP) to confirm incorporation of the rpoB-GFPUV construct. Cultures were then analyzed by flow cytometry for detection of GFPUV expression, and transformed R. typhi were isolated in a fluorescence activated cell sorter. This is the first report of transformation of rickettsiae with a nonrickettsial (GFPUV) gene. PMID- 10377107 TI - Histoplasma capsulatum strain variation in both H antigen production and beta glucosidase activity and overexpression of HAG1 from a telomeric linear plasmid. AB - The H antigen of the dimorphic fungal pathogen Histoplasma capsulatum was first described over 40 years ago. It is a secreted glycoprotein that is immunogenic during infection. Recent cloning of the H antigen gene (HAG1) indicated sequence homology with genes for fungal beta-glucosidases. To understand the biological role of this immunodominant antigen in H. capsulatum, enzymatic assays were performed to determine whether H. capsulatum contained a beta-glucosidase enzyme activity and whether this activity was encoded by the HAG1 gene. Substrate gels with H. capsulatum culture supernatants revealed beta-glucosidase activity near the predicted mobility of the H antigen. Quantitative microtiter plate assays revealed marked differences in secreted beta-glucosidase activities from three H. capsulatum restriction fragment length polymorphism (RFLP) classes, with RFLP class II strains displaying high levels of enzyme activity, in contrast to the low levels of activity exhibited by class I and III strains. Immunoblotting of culture supernatants with an H antigen-specific antiserum demonstrated differences in H protein expression levels between the H. capsulatum classes, with a correlation between secreted enzyme activity and H protein levels. We took advantage of these class differences to demonstrate multicopy plasmid H gene overexpression by transformation of an HAG1 plasmid into H. capsulatum. Both a class II strain (G217Bura5-23) and a class III strain (G184ASura5-11) transformed with the telomeric overexpression plasmid pMAD401 displayed increased levels of beta-glucosidase enzyme activity and H protein expression compared to the levels in control transformants containing only the single genomic copy of HAG1. This is the first demonstration of telomeric plasmid-mediated protein overexpression in this pathogenic fungus, and the findings support the identification of the H antigen as a beta-glucosidase. PMID- 10377108 TI - Prevalence of, antibody response to, and immunity induced by Haemophilus ducreyi hemolysin. AB - Haemophilus ducreyi, the etiologic agent of chancroid, a genital ulcer disease, produces a cell-associated hemolysin whose role in virulence is not well defined. Hemolysin is encoded by two genes, hhdA and hhdB, which, based on their homology to Serratia marcescens shlA and shlB genes, are believed to encode the hemolysin structural protein and a protein required for secretion and modification of this protein, respectively. In this study, we determined the prevalence and expression of the hemolysin genes in 90 H. ducreyi isolates obtained from diverse geographic locations from 1952 to 1996 and found that all strains contained DNA homologous to the hhdB and hhdA genes. In addition, all strains expressed a hemolytic activity. We also determined that hemolysin is expressed in vivo and is immunogenic, as indicated by the induction of antibodies to hemolysin in both the primate and rabbit disease models as well as in human patients with naturally acquired chancroid. Wild-type strain 35000 and isogenic hemolysin-negative mutants showed no difference in lesion development in the temperature-dependent rabbit model. However, immunization of rabbits with the purified hemolysin protein reduced the recovery of wild-type H. ducreyi, but not hemolysin-negative mutants, from lesions. Our study indicates that hemolysin is a possible candidate for vaccine development due to its immunogenicity, expression in vitro and in vivo by most, if not all, strains, and the effect of immunization on reducing the recovery of viable H. ducreyi in experimental disease in rabbits. PMID- 10377109 TI - Experimental lyme arthritis in the absence of interleukin-4 or gamma interferon. AB - Genetic resistance and susceptibility to experimental Lyme arthritis have been linked with the production of interleukin-4 (IL-4) or gamma interferon (IFN gamma), respectively. To determine the absolute requirement for these cytokines in disease outcome, we compared arthritis development in wild-type, IL-4 deficient (IL-4 degrees ), and IFN-gamma-deficient (IFN-gamma degrees ) mice. While susceptible C3H mice developed swelling of ankle joints during the second week of infection, this swelling was exacerbated in C3H IFN-gamma degrees mice. Their arthritis severity scores at day 21, however, were similar. Resolution of arthritis was also similar between C3H and C3H IFN-gamma degrees mice. Arthritis resistant DBA mice did not develop ankle swelling during the experimental period. There were no differences in ankle swelling or arthritis severity scores between control DBA mice and DBA IL-4 degrees mice at any of the time points tested. While the presence of spirochetes in various tissues was similar among all strains at day 21, DBA IL-4 degrees mice had a higher presence of spirochetes in blood, heart, and spleen than the DBA, C3H, and C3H IFN-gamma degrees mice did at day 60. DBA IL-4 degrees mice also had impaired ability to produce Borrelia specific antibody responses, especially immunoglobulin G1. Thus, while IFN-gamma and IL-4 are not absolutely required for arthritis susceptibility or resistance, the production of IL-4 does appear to play an important role in Borrelia-specific antibody production and spirochete clearance. PMID- 10377110 TI - T-Cell responses during Trypanosoma brucei infections in mice deficient in inducible nitric oxide synthase. AB - We have investigated the possibility that nitric oxide (NO) synthesis may affect the course of a trypanosome infection via T-cell responses using mice deficient in inducible NO synthase (iNOS). Parasitemia levels increased at the same rate in both iNOS-deficient homozygous and control heterozygous mice, and peak parasitemia values were the same in both groups. However, the heterozygous mice maintained higher parasitemia levels after the peak of an infection than the homozygous mice due to a decrease in the rate of clearance of parasites. In iNOS deficient mice there was an increase in the numbers of total CD4(+) cells and activated (interleukin-2 receptor-expressing) CD4(+) cells in infected mice compared with the numbers in uninfected mice. Spleen cells from infected iNOS deficient mice displayed increased proliferative responses and gamma interferon secretion when stimulated in vitro than those of control mice. These data suggest that NO production depresses T-helper 1-like responses generated during Trypanosoma brucei infections, thus promoting the survival of the parasite. PMID- 10377111 TI - A novel means of self-protection, unrelated to toxin activation, confers immunity to the bactericidal effects of the Enterococcus faecalis cytolysin. AB - Enterococcus faecalis has become a pervasive clinical problem due to the emergence of resistance to most antibiotics. The cytolysin of E. faecalis is a novel bacterial toxin that contributes to the severity of disease. It consists of two structural subunits, which together possess both hemolytic and bactericidal activity. Both toxin subunits are encoded in a complex operon frequently harbored on pheromone-responsive plasmids. E. faecalis strains lacking such plasmids are susceptible to the bactericidal effects of the cytolysin. A novel cytolysin immunity determinant at the 3' end of the pAD1 cytolysin operon is described in the present study. Deletion analysis and specific mutagenesis isolated the immunity function to a single open reading frame. Specific mutagenesis experiments demonstrate that cytolysin immunity is unrelated to cytolysin activator (CylA) expression as previously proposed. Cytolysin immunity is, however, encoded on the same transcript as and 3' to CylA, and previous associations between immunity and CylA can be ascribed to the polar behavior of Tn917 insertion. PMID- 10377113 TI - Role of hemolysin BL in the pathogenesis of extraintestinal Bacillus cereus infection assessed in an endophthalmitis model. AB - Bacillus cereus is a rare cause of serious human infection but, paradoxically, causes one of the most severe posttraumatic or endogenous infections of the eye, endophthalmitis, which frequently results in blindness. The virulence of B. cereus endophthalmitis historically has been attributed to toxin production. We therefore sought to examine the contribution of the dermonecrotic toxin, hemolysin BL, to the pathogenesis of B. cereus infection in an endophthalmitis system that is highly amenable to study. The pathogenesis of infection resulting from intravitreal injection of 10(2) CFU of either a clinical ocular isolate of B. cereus producing hemolysin BL (HBL+) or an isogenic mutant in this trait (HBL ) was assessed bacteriologically and by slit lamp biomicroscopy, electroretinography, histology, and inflammatory cell enumeration. Both HBL+ and HBL- strains evoked severe intraocular inflammatory responses as early as 12 h postinfection, with complete loss of retinal responsiveness by 12 h. The infections caused by both strains spread of the infection to adjacent tissues by 18 h. No significant differences in intraocular bacterial growth (P >/= 0.21) or inflammatory changes (P >/= 0.21) were observed in eyes infected with either HBL+ or HBL- strains during the course of infection. The level of retinal responsiveness was greater in HBL- infected eyes than in HBL+-infected eyes at 6 h only (P = 0.01). These results indicate that hemolysin BL makes no essential contribution to the severe and rapid course of infection in the endophthalmitis model. PMID- 10377112 TI - Pathogenesis of gram-positive bacterial endophthalmitis. AB - The severity of endophthalmitis has been associated generally with the virulence of the offending pathogen. However, precisely what constitutes the virulence in intraocular infections remains ill defined. We therefore sought to identify the basis for virulence for three common ocular pathogens (Bacillus cereus, Enterococcus faecalis, and Staphylococcus aureus) in terms of intraocular growth rates, bacterial localization patterns, and the contribution of cell walls and secreted products to the pathogenesis of endophthalmitis. Rabbit eyes were injected intravitreally with (i) viable B. cereus, E. faecalis, or S. aureus, (ii) metabolically inactive B. cereus, E. faecalis, or S. aureus, (iii) sacculus preparations from each strain, or (iv) culture fluid containing products secreted by each strain. Eyes were assessed at various times following injection by slit lamp biomicroscopy, electroretinography (ERG), bacterial and inflammatory cell enumeration, and histology. B. cereus endophthalmitis followed a more rapid and virulent course than E. faecalis or S. aureus endophthalmitis, eliminating retinal responsiveness, as measured by ERG, by 12 h. Analysis of bacterial localization revealed that B. cereus uniquely migrated rapidly from posterior to anterior segment during infection. Although injection of neither metabolically inactive bacteria nor cell wall sacculi greatly affected ERG, significant intraocular inflammation was observed. Injection of B. cereus or S. aureus culture fluids caused both significant reductions in retinal responsiveness and significant intraocular inflammation, paralleling that seen in natural infections. The results demonstrate that toxins, intraocular localization, and, to a lesser extent, the intraocular host response to cell walls all contribute to the pathogenesis of B. cereus, S. aureus, and E. faecalis endophthalmitis in a pathogen-specific manner. The key pathophysiologic differences in these intraocular diseases highlight opportunities for optimizing conventional therapies and deriving new ones. PMID- 10377114 TI - Role of superoxide dismutase activity in the physiology of Porphyromonas gingivalis. AB - Porphyromonas gingivalis is a gram-negative, obligate anaerobe strongly associated with chronic adult periodontitis. A previous study has demonstrated that this organism requires superoxide dismutase (SOD) for its modest aerotolerance. In this study, we have constructed a mutant deficient in SOD activity by insertional inactivation as well as a sod::lacZ reporter translational fusion construct to study the regulation of expression of this gene. We have confirmed that SOD is essential for tolerance to atmospheric oxygen but does not appear to be protective against hydrogen peroxide or exogenously generated reactive oxygen species. Furthermore, the sod mutant appeared to be no more sensitive to killing by neutrophils than the parental strain 381. SOD appears to be protective against oxygen-dependent DNA damage as measured by increased mutation to rifampin resistance by the sod mutant. Use of the sod::lacZ construct confirmed that SOD expression is maximal at mid-log phase and is influenced by oxygen, temperature, and pH. However, expression does not appear to be significantly affected by iron depletion, osmolarity, or nutrient depletion. The transcription start site of the sod gene was determined to be 315 bp upstream of the sod start codon and to be within an upstream open reading frame. Our studies demonstrate the essential role that SOD plays in aerotolerance of this organism as well as the selective induction of this enzyme by environmental stimuli. PMID- 10377115 TI - Deacylation of purified lipopolysaccharides by cellular and extracellular components of a sterile rabbit peritoneal inflammatory exudate. AB - The extent to which the mammalian host is capable of enzymatic degradation and detoxification of bacterial lipopolysaccharides (LPS) is still unknown. Partial deacylation of LPS by the enzyme acyloxyacyl hydrolase (AOAH) provides such a mechanism, but its participation in the disposal of LPS under physiological conditions has not been established. In this study, deacylation of isolated radiolabeled LPS by both cellular and extracellular components of a sterile inflammatory peritoneal exudate elicited in rabbits was examined ex vivo. AOAH like activity, tested under artificial conditions (pH 5.4, 0.1% Triton X-100), was evident in all components of the exudate (mononuclear cells [MNC] > polymorphonuclear leukocytes [PMN] > inflammatory [ascitic] fluid [AF]). Under more physiological conditions, in a defined medium containing purified LPS binding protein, the LPS-deacylating activity of MNC greatly exceeded that of PMN. In AF, MNC (but not PMN) also produced rapid and extensive CD14-dependent LPS deacylation. Under these conditions, almost all MNC-associated LPS underwent deacylation within 1 h, a rate greatly exceeding that previously found in any cell type. The remaining extracellular LPS was more slowly subject to CD14 independent deacylation in AF. Quantitative analysis showed a comparable release of laurate and myristate but no release of 3-hydroxymyristate, consistent with an AOAH-like activity. These findings suggest a major role for CD14(+) MNC and a secondary role for AF in the deacylation of cell-free LPS at extravascular inflammatory sites. PMID- 10377117 TI - Mycoplasmal lipopeptide MALP-2 induces the chemoattractant proteins macrophage inflammatory protein 1alpha (MIP-1alpha), monocyte chemoattractant protein 1, and MIP-2 and promotes leukocyte infiltration in mice. AB - Natural as well as experimental infections with pathogenic mycoplasmas lead to cellular responses characterized by early polymorphonuclear leukocyte influx, which in turn is followed by infiltration of macrophages. Since some of the most potent leukocyte chemoattractants are macrophage products, we investigated whether the 2-kDa macrophage-activating lipopeptide (MALP-2) from Mycoplasma fermentans was capable of inducing chemoattractant chemokines and initiating an in vivo inflammatory effect. MALP-2 was a potent in vitro inducer of the chemokines macrophage inflammatory protein 1alpha (MIP-1alpha), monocyte chemoattractant protein 1 (MCP-1), and MIP-2, yielding a maximal response at 0.1 ng/ml (5 x 10(-11) M). Leukocyte infiltration was determined after intraperitoneal injection of MALP-2, liposome-encapsulated MALP-2, and heat killed mycoplasmas. There was a steady increase in the number of peritoneal cells over 72 h in response to these agents. Polymorph counts were maximal by 24 to 48 h, decreasing thereafter. Monocytes/macrophages had significantly increased after 3 days. MIP-1alpha, MCP-1, and MIP-2 levels in serum or peritoneal lavage fluid were determined. MIP-1alpha and MCP-1 levels were elevated by 2 to 6 h after injection and were still above control values after 24 h. In contrast, MIP-2 levels reached their maximum at 2 h, dropping to control values after 24 h. We conclude that macrophage-stimulating mycoplasmal lipoproteins, exemplified by MALP-2, play an important role in the late phase of phagocyte recruitment at sites of infection and that this is affected by leukoattractive chemokines. PMID- 10377116 TI - Isolation of T-cell antigens by using a recombinant protein library and its application to the identification of novel vaccine candidates against schistosomiasis. AB - We present here a novel approach to identify T-cell antigens from any infectious agent by use of a library of purified recombinant proteins. Essential features of this strategy include (i) a highly efficient cDNA cloning system which negatively selects against nonrecombinant transformants by making use of the bacterial EcoK restriction system, (ii) affinity staining of cDNA clones expressing recombinant proteins, and (iii) a procedure of simultaneous purification of recombinant proteins from large numbers of isolated clones (representing the protein library) in a single step from pools consisting of up to 24 individual clones. The feasibility of the screening system was confirmed by constructing a protein library of the human parasite Schistosoma mansoni. The recombinant antigens of this library were used to stimulate CD4(+) T cells derived from the axillary lymph nodes of mice vaccinated with irradiated cercariae. In initial screening experiments, we detected parasite-specific proliferation and gamma interferon (IFN-gamma) secretion in response to several pools of cDNA clones. Further analysis of one particular pool revealed that only one of its constituents stimulated considerable IFN-gamma secretion by CD4(+) T cells and that the expressed antigen is identical to a small fragment of myosin heavy chain. PMID- 10377118 TI - Induction of lethal shock and tolerance by Porphyromonas gingivalis lipopolysaccharide in D-galactosamine-sensitized C3H/HeJ mice. AB - Lipopolysaccharide (LPS) obtained from Porphyromonas gingivalis was found to exhibit marked lethal toxicity in galactosamine-sensitized C3H/HeJ mice. Although no lethality was observed in mice intraperitoneally challenged with 1 mg of P. gingivalis LPS without galactosamine, when they were sensitized with 30 mg of galactosamine, challenge with 1 and 10 micrograms of LPS resulted in 67 and 100% lethality, respectively. The lethal dose of LPS was almost the same in LPS responsive C57BL/6 mice and non-LPS-responsive C3H/HeJ mice. Furthermore, when 1 microgram of P. gingivalis LPS was administered to each mouse 90 min before the challenge with the same LPS with galactosamine, tolerance to the lethal action of LPS was induced, and the mice were completely protected from death, even at a dose 100-fold greater than the lethal dose of LPS. Neither a lethal effect nor induction of tolerance to the lethality of P. gingivalis LPS was exhibited by Salmonella LPS in galactosamine-sensitized C3H/HeJ mice. A protein-LPS complex derived from Pseudomonas aeruginosa, which exhibited strong lethality and induced tolerance to a subsequent challenge with a lethal dose of LPS in galactosamine sensitized LPS-responsive mice, did not exhibit lethal toxicity in galactosamine sensitized C3H/HeJ mice and failed to induce tolerance in these mice to the lethality of P. gingivalis LPS. These results indicate that P. gingivalis LPS plays the central role in the activation of non-LPS-responsive C3H/HeJ mice. PMID- 10377119 TI - Schistosoma mansoni activates host microvascular endothelial cells to acquire an anti-inflammatory phenotype. AB - Since endothelial cells (ECs) play a key role in immune defense mechanisms and in immunopathology, we investigated whether the intravascular helminth parasite Schistosoma mansoni could interact with and activate resting ECs in vitro. Microscopic analysis revealed that the lung-stage schistosomula specifically attached to microvascular ECs. This adherence was associated to active cellular processes involving actin filament formation. Since variation of permeability of cultured capillary brain ECs is a good marker for endothelial activation, the transendothelial passage of a low-molecular-weight molecule (inulin) on monolayers of bovine brain capillary ECs (BBCEC) was measured in response to parasites. Schistosomula induced a dramatic decrease in transendothelial permeability, a characteristic marker for the generation of an anti-inflammatory phenotype to ECs. This paracellular barrier enhancing effect on endothelial monolayers was due to a soluble substance(s) (below 1 kDa in size) secreted from S. mansoni schistosomula and not by mechanisms associated to adherence between parasites and ECs. The reinforcement of the endothelial barrier function was accompanied by an elevation of intracellular concentration of cyclic AMP (cAMP). The use of specific kinase inhibitors confirms that schistosomula activate ECs through a cAMP/protein kinase A pathway that leads to an increased phosphorylation of the myosin light-chain kinase. These combined findings suggest that the secretory/excretory products from schistosomula possess anti inflammatory factor(s) that signal host microvascular endothelium. The immunological consequences of such activation are discussed. PMID- 10377120 TI - A localized adherence-like pattern as a second pattern of adherence of classic enteropathogenic Escherichia coli to HEp-2 cells that is associated with infantile diarrhea. AB - Escherichia coli strains that cause nonbloody diarrhea in infants are known to present three distinct patterns of adherence to epithelial cells, namely, localized (LA), diffuse (DA), and aggregative (AA) adherence. Strains with LA (typical Enteropathogenic Escherichia coli [EPEC]) are well recognized as a cause of secretory diarrhea, but the role of strains with DA (DAEC) is controversial, and strains with AA (EAEC) have been more frequently related to persistent diarrhea whereas its relationship with acute diarrhea is not well defined. To determine the relationship of the different types of E. coli adherence patterns with acute diarrhea (lasting less than 14 days) and persistent diarrhea (lasting more than 14 days) in Sao Paulo, Brazil, we studied stool specimens from 40 infants under 1 year of age with diarrhea and 40 age-matched control infants without any gastrointestinal symptoms. Twenty-eight (35.0%) of eighty cases yielded adherent E. coli (HEp-2 cells). Strains with localized and aggregative adherence were associated with acute and persistent diarrhea. A total of 11.2% of the adherent strains were typical EPEC serotypes and hybridized with the enteroadherence factor probe; 5.0% were EAEC and hybridized with the EAEC probe. DAEC strains were isolated from 10.0% of patients and 7.5% of controls and did not hybridize with the two probes used (daaC and AIDA-I). Strains with a localized adherence-like pattern (atypical EPEC) were found significantly more frequently (P = 0.028) in cultures from children with diarrhea (17.5%) than in controls (2.5%). PMID- 10377121 TI - Genomic loci of the Porphyromonas gingivalis insertion element IS1126. AB - The Porphyromonas gingivalis genome contains multiple copies of insertion element IS1126. When chromosomal DNA digests of different strains were probed with IS1126, between 25 and 35 hybridizing fragments per genome were detected, depending on the strain. Unrelated strains had very different restriction fragment length polymorphism (RFLP) patterns. When different laboratory copies of a specific strain were examined, the IS1126 RFLP patterns were very similar but small differences were observed, indicating that element-associated changes had occurred during laboratory passage. Within the next year, genome sequencing, assembly, and annotation for P. gingivalis W83 will be completed. Because repetitive elements complicate the assembly of randomly sequenced DNA fragments, we isolated and sequenced the flanking regions of IS1126 copies in strain W83. We also isolated and sequenced the flanking regions of IS1126 copies in strain ATCC 33277 in order to compare insertion sites in phylogenetically divergent strains. We identified 37 new sequences flanking IS1126 from strain ATCC 33277 and 30 from strain W83. The insertion element was found between genes except where it transposed into another insertion element. Examination of identifiable flanking genes or open reading frames indicated that the insertion sites were different in the two strains, except that both strains possess an insertion adjacent to the Lys-gingipain gene (J. P. Lewis and F. L. Macrina, Infect. Immun. 66:3035-3042, 1998). Most of the genes or sequences flanking IS1126 in ATCC 33277 were present in W83 but were contiguous and not insertion element associated. Thus, where genes were identified in both strains, their order was maintained, indicating that the two genomes are organized similarly, but the loci of IS1126 are different. In both strains, insertion element-associated duplicated target sites were lost from several copies of IS1126, providing evidence of homologous recombination between elements. Larger organizational differences between the genomes, such as deletions and inversions, may result from insertion element mediated recombination events. PMID- 10377122 TI - Interleukin-10 responses to liver-stage antigen 1 predict human resistance to Plasmodium falciparum. AB - The design of an effective vaccine against Plasmodium falciparum, the most deadly malaria parasite of humans, requires a careful definition of the epitopes and the immune responses involved in protection. Liver-stage antigen 1 (LSA-1) is specifically expressed during the hepatic stage of P. falciparum and elicits cellular and humoral immune responses in naturally exposed individuals. We report here that interleukin-10 (IL-10) production in response to LSA-1 predicts resistance to P. falciparum after eradication therapy. Resistance was not related to gamma interferon or tumor necrosis factor alpha production. This is the first report that human IL-10 responses are associated with resistance after eradication therapy, and our findings support the inclusion of LSA-1 in a vaccine against malaria. PMID- 10377123 TI - Pretreatment with granulocyte colony-stimulating factor attenuates the inflammatory response but not the bacterial load in cerebrospinal fluid during experimental pneumococcal meningitis in rabbits. AB - A possible immunomodulatory role of granulocyte colony-stimulating factor (G-CSF) was investigated in an experimental pneumococcal meningitis model in rabbits. Animals were pretreated with G-CSF (10 micrograms/kg subcutaneously twice a day) starting 48 h before in vivo and ex vivo experiments, causing a five- to six-fold increase in the peripheral leukocyte level. Meningitis was induced by intracisternal inoculation of approximately 4 x 10(5) CFU of Streptococcus pneumoniae type 3. Neutrophil pleocytosis and interleukin-8 (IL-8) levels were significantly attenuated in G-CSF-pretreated animals compared to untreated animals (P < 0.05). Furthermore, G-CSF pretreatment significantly delayed alterations in cerebrospinal fluid (CSF) tumor necrosis factor alpha and IL-1beta levels, as well as protein and glucose levels (P < 0.05). No difference in CSF bacterial concentrations was found, whereas the blood bacterial concentration was significantly decreased in G-CSF-pretreated animals (P < 0.05). Ex vivo chemotaxis of neutrophils isolated from G-CSF-pretreated animals was significantly decreased compared to that of neutrophils from untreated animals (P < 0.05). In conclusion, G-CSF pretreatment attenuates meningeal inflammation and enhances systemic bacterial killing. Further preclinical studies are required to investigate whether this may affect the clinical course of meningitis and thus whether G-CSF treatment may have a beneficial role in pneumococcal meningitis. PMID- 10377124 TI - Vaccination against shigellosis with attenuated Shigella flexneri 2a strain SC602. AB - The Shigella flexneri 2a SC602 vaccine candidate carries deletions of the plasmid borne virulence gene icsA (mediating intra- and intercellular spread) and the chromosomal locus iuc (encoding aerobactin) (S. Barzu, A. Fontaine, P. J. Sansonetti, and A. Phalipon, Infect. Immun. 64:1190-1196, 1996). Dose selection studies showed that SC602 causes shigellosis in a majority of volunteers when 3 x 10(8) or 2 x 10(6) CFU are ingested. In contrast, a dose of 10(4) CFU was associated with transient fever or mild diarrhea in 2 of 15 volunteers. All volunteers receiving single doses of >/=10(4) CFU excreted S. flexneri 2a, and this colonization induced significant antibody-secreting cell and enzyme-linked immunosorbent assay responses against S. flexneri 2a lipopolysaccharide in two thirds of the vaccinees. Seven volunteers who had been vaccinated 8 weeks earlier with a single dose of 10(4) CFU and 7 control subjects were challenged with 2 x 10(3) CFU of virulent S. flexneri 2a organisms. Six of the control volunteers developed shigellosis with fever and severe diarrhea or dysentery, while none of the vaccinees had fever, dysentery, or severe symptoms (P = 0. 005). Three vaccinees experienced mild diarrhea, and these subjects had lower antibody titers than did the fully protected volunteers. Although the apparent window of safety is narrow, SC602 is the first example of an attenuated S. flexneri 2a candidate vaccine that provides protection against shigellosis in a stringent, human challenge model. PMID- 10377126 TI - Chemokine gene expression during Pneumocystis carinii-driven pulmonary inflammation. AB - Severe combined immunodeficient (SCID) mice lack functional lymphocytes and therefore develop Pneumocystis carinii pneumonia. However, when infected SCID mice are immunologically reconstituted with congenic spleen cells, a protective inflammatory cascade is initiated. Proinflammatory cytokines are produced, and lymphocytes and macrophages are recruited specifically to alveolar sites of infection. Importantly, uninfected regions of the lung remain free from inflammatory involvement, suggesting that there are specific mechanisms that limit inflammation in the infected lung. Therefore, to determine whether chemokines are involved in targeting the P. carinii-driven inflammatory response, steady-state mRNA levels of several chemokines were measured in the lungs of both reconstituted and nonreconstituted P. carinii-infected SCID mice. Despite significant organism burdens in the lungs of 8- and 10-week-old SCID mice, there was no evidence of elevated chemokine gene expression, which is consistent with the lack of an inflammatory response in these animals. However, when 8-week-old infected SCID mice were immunologically reconstituted, signs of focal pulmonary inflammation were observed, and levels of RANTES, MCP-1, lymphotactin, MIP 1alpha, MIP-1beta, and MIP-2 mRNAs were all significantly elevated. Chemokine mRNA abundance was elevated at day 10 postreconstitution (PR), was maximal at day 12 PR, and returned to baseline by day 22 PR. In situ hybridization demonstrated that during the peak of inflammation, RANTES gene expression was localized to sites of inflammatory cell infiltration and P. carinii infection. Thus, these observations indicate that chemokines play a role in the focal targeting of inflammatory cell recruitment to sites of P. carinii infection after the passive transfer of lymphocytes to the host. PMID- 10377125 TI - Development of antibody isotype responses to Schistosoma mansoni in an immunologically naive immigrant population: influence of infection duration, infection intensity, and host age. AB - We have identified the influence of host and parasite factors that give rise to characteristic antibody isotype profiles with age seen in human populations living in different areas of schistosomiasis endemicity. This is important in the immunobiology of this disease. It is also of interest in the context of human responses to chronic antigen stimulation, vaccines, allergens, and other pathogens. In populations exposed to endemic schistosomiasis, factors such as intensity and duration of infection are age dependent. They therefore confound the influence of host age on antiparasite responses. Here, we resolved these confounding factors by comparing the developing antibody responses of an immunologically naive immigrant population as they acquired the infection for the first time with those of chronically infected resident inhabitants of the same region of Schistosoma mansoni endemicity in Kenya. Recent arrival in the area strongly favored immunoglobulin G3 (IgG3) responses against the parasite. The antibody isotype responses associated with human susceptibility to reinfection after chemotherapy were elevated in those suffering high intensities of infection (IgG4 responses against worm and egg antigens) or were characteristic responses of young children irrespective of the intensity or duration of infection (IgG2 responses against egg antigen). IgE responses against the adult worm, a response associated with resistance to reinfection after chemotherapy, increased with the ages of infected individuals and were also favored in those currently suffering higher intensities of infection. PMID- 10377127 TI - Wheat germ agglutinin induces NADPH-oxidase activity in human neutrophils by interaction with mobilizable receptors. AB - Wheat germ agglutinin (WGA), a lectin with specificity for N-acetylglucosamine and sialic acid, was investigated with respect to its ability to activate the NADPH-oxidase of in vivo-exudated neutrophils (obtained from a skin chamber), and the activity was compared to that of peripheral blood neutrophils. The exudate cells responded to WGA, by both releasing reactive oxygen species into the extracellular milieu and producing oxygen metabolites intracellularly. The peripheral blood cells were unresponsive. To mimic the in vivo-exuded neutrophils with regards to receptor exposure, peripheral blood neutrophils were induced to mobilize their granules and vesicles to varying degrees (in vitro priming), prior to challenge with WGA. The oxidative response to WGA increased with increasing levels of granule mobilization, and the receptor(s) could be shown to reside in the secretory vesicles and/or the gelatinase granules in resting neutrophils. Several WGA-binding glycoproteins were detected in subcellular fractions containing these organelles. The extra- and intracellular NADPH-oxidase responses showed differences in sialic acid dependency, indicating that these two responses are mediated by different receptor structures. PMID- 10377128 TI - Neisseria gonorrhoeae coordinately uses Pili and Opa to activate HEC-1-B cell microvilli, which causes engulfment of the gonococci. AB - This study was undertaken to examine concomitant roles of pili and colony opacity associated proteins (Opa) in promoting Neisseria gonorrhoeae adherence to and invasion of human endometrial HEC-1-B cells. Adherence of N. gonorrhoeae to cultured HEC-1-B cells was saturable, even though organisms adhered to <50% of the cells. During 4 to 6 h of incubation, adherent mono- and diplococci formed microcolonies on the surfaces of the cells. Microvilli of the HEC-1-B cells adhered by their distal ends to individual cocci within the microcolonies. When the microcolonies grew from isogenic pilus-negative (P-) Opa-, P- Opa+, or P+ Opa gonococci, microvilli did not elongate, and the colonies were not engulfed. In contrast, the microvilli markedly elongated during exposure to P+ Opa+ gonococci. The microvilli adhered to the organisms along their full lengths and appeared to actively participate in the engulfment of the microcolonies. Internalized microcolonies, with P+ Opa+ gonococci, contained dividing cocci and appeared to be surrounded by cell membrane but were not clearly within vacuoles. In contrast, degenerate individual organisms were within vacuoles. Low doses of chloramphenicol, which inhibits protein synthesis by both prokaryotes and eukaryotes, prevented the microvillar response to and internalization of the P+ Opa+ gonococci; higher doses caused internalization without microvillus activation. Cycloheximide and anisomycin, which inhibit only eukaryotic protein synthesis, caused dose-dependent enhancement of uptake. Cytochalasins reduced engulfment; colchicine had no effect. These results show that gonococci must express both pili and Opa to be engulfed efficiently by HEC-1-B cells. PMID- 10377129 TI - Biased immunoglobulin G1 isotype responses induced in cattle with DNA expressing msp1a of Anaplasma marginale. AB - Immunization with the native major surface protein 1 (MSP1) (a heterodimer containing disulfide and noncovalently bonded polypeptides designated MSP1a and MSP1b) of the erythrocytic stage of Anaplasma marginale conferred protection against homologous challenge (G. H. Palmer, A. F. Barbet, W. C. Davis, and T. C. McGuire, Science 231:1299-1302, 1986). The MSP1a polypeptide possesses a conserved neutralization-sensitive epitope. In the present study, the immune response to DNA-mediated immunization using msp1a was studied. The plasmid pVCL/MSP1a, which encodes the complete msp1a gene of A. marginale under the control of human cytomegalovirus immediate-early enhancer/promoter and intron A, was constructed. The immune responses elicited by immunization with pVCL/MSP1a into cardiotoxin-induced regenerating muscle were evaluated in mice and cattle. Antibody reactive with native MSP1a was detected in pooled sera of immunized BALB/c mice 3 weeks following primary immunization. Two calves seronegative for A. marginale were immunized four times, at weeks 0, 3, 7, and 13, with pVCL/MSP1a. By 8 weeks, both calves responded to MSP1a with an antibody titer of 1:100, which peaked at 1:1,600 and 1:800 by 16 weeks after the initial immunization. Interestingly, immunoblotting with anti-immunoglobulin G1 (anti IgG1) and anti-IgG2 specific monoclonal antibodies revealed a restricted IgG1 anti-MSP1a response in both animals. T-lymphocyte lines, established after the fourth immunization, proliferated specifically against A. marginale homogenate and purified MSP1 in a dose-dependent manner. These data provide a basis for an immunization strategy to direct bovine immune responses by using DNA vaccine vectors containing single or multiple genes encoding major surface proteins of A. marginale. PMID- 10377130 TI - Effect of peroxisome proliferator-activated receptor alpha activators on tumor necrosis factor expression in mice during endotoxemia. AB - Inflammatory mediators orchestrate the host immune and metabolic response to acute bacterial infections and mediate the events leading to septic shock. Tumor necrosis factor (TNF) has long been identified as one of the proximal mediators of endotoxin action. Recent studies have implicated peroxisome proliferator activated receptor alpha (PPARalpha) as a potential target to modulate regulation of the immune response. Since PPARalpha activators, which are hypolipidemic drugs, are being prescribed for a significant population of older patients, it is important to determine the impact of these drugs on the host response to acute inflammation. Therefore, we examined the role of PPARalpha activators on the regulation of TNF expression in a mouse model of endotoxemia. CD-1 mice treated with dietary fenofibrate or Wy-14,643 had fivefold-higher lipopolysaccharide (LPS)-induced TNF plasma levels than LPS-treated control-fed animals. Higher LPS induced TNF levels in drug-fed animals were reflected physiologically in significantly lower glucose levels in plasma and a significantly lower 50% lethal dose than those in LPS-treated control-fed animals. Utilizing PPARalpha wild-type (WT) and knockout (KO) mice, we showed that the effect of fenofibrate on LPS induced TNF expression was indeed mediated by PPARalpha. PPARalpha WT mice fed fenofibrate also had a fivefold increase in LPS-induced TNF levels in plasma compared to control-fed animals. However, LPS-induced TNF levels were significantly decreased and glucose levels in plasma were significantly increased in PPARalpha KO mice fed fenofibrate compared to those in control-fed animals. Data from peritoneal macrophage studies indicate that Wy-14,643 modestly decreased TNF expression in vitro. Similarly, overexpression of PPARalpha in 293T cells decreased activity of a human TNF promoter-luciferase construct. The results from these studies suggest that any anti-inflammatory activity of PPARalpha in vivo can be masked by other systemic effects of PPARalpha activators. PMID- 10377131 TI - Differential sensitivity of human epithelial cells to Pseudomonas aeruginosa exoenzyme S. AB - Exoenzyme S (ExoS) is an ADP-ribosyltransferase produced and directly translocated into eukaryotic cells by the opportunistic pathogen Pseudomonas aeruginosa. Model systems that allow bacterial translocation of ExoS have found ExoS to have multiple effects on eukaryotic cell function, affecting DNA synthesis, actin cytoskeletal structure, and cell matrix adherence. To understand mechanisms underlying differences observed in cell sensitivities to ExoS, we examined the effects of bacterially translocated ExoS on multiple human epithelial cell lines. Of the cell lines examined, confluent normal kidney (NK) epithelial cells were most resistant to ExoS, while tumor-derived cell lines were highly sensitive to ExoS. Analysis of the mechanisms of resistance indicated that cell association as well as an intrinsic resistance to morphological alterations were associated with increased resistance to ExoS. Conversely, increased sensitivity to ExoS appeared to be linked to epithelial cell growth, with tumor cells capable of undergoing non-contact-inhibited, anchorage-independent growth all being sensitive to ExoS, and NK cells becoming sensitive to ExoS when subconfluent and growing. Consistent with the possibility that growth-related, actin-based structures are involved in sensitivity to ExoS, scanning electron microscopy revealed cellular extensions from sensitive, growing cells to bacteria, which were not readily evident in resistant cells. In all studies, the severity of effects of ExoS on cell function directly correlated with the degree of Ras modification, indicating that sensitivity to ExoS in some manner related to the efficiency of ExoS translocation and its ADP-ribosylation of Ras. Our results suggest that factors expressed by growing epithelial cells are required for the bacterial contact-dependent translocation of ExoS; as normal epithelial cells differentiate into polarized confluent monolayers, expression of these factors is altered, and cells in turn become more resistant to the effects of ExoS. PMID- 10377132 TI - Differential roles of segmented filamentous bacteria and clostridia in development of the intestinal immune system. AB - The presence of microflora in the digestive tract promotes the development of the intestinal immune system. In this study, to evaluate the roles of two types of indigenous microbe, segmented filamentous bacteria (SFB) and clostridia, whose habitats are the small and large intestines, respectively, in this immunological development, we analyzed three kinds of gnotobiotic mice contaminated with SFB, clostridia, and both SFB and clostridia, respectively, in comparison with germfree (GF) or conventionalized (Cvd) mice associated with specific-pathogen free flora. In the small intestine, the number of alpha beta T-cell receptor bearing intraepithelial lymphocytes (alpha betaIEL) increased in SFB-associated mice (SFB-mice) but not in clostridium-associated mice (Clost-mice). There was no great difference in Vbeta usage among GF mice, Cvd mice, and these gnotobiotic mice, although the association with SFB decreased the proportion of Vbeta6(+) cells in CD8beta- subsets to some extent, compared to that in GF mice. The expression of major histocompatibility complex class II molecules on the epithelial cells was observed in SFB-mice but not in Clost-mice. On the other hand, in the large intestine, the ratio of the number of CD4(-) CD8(+) cells to that of CD4(+) CD8(-) cells in alpha betaIEL increased in Clost-mice but not in SFB-mice. On association with both SFB and clostridia, the numbers and phenotypes of IEL in the small and large intestines changed to become similar to those in Cvd mice. In particular, the ratio of the number of CD8alpha beta+ cells to that of CD8alpha alpha+ cells in alpha betaIEL, unusually elevated in the small intestines of SFB-mice, decreased to the level in Cvd mice on contamination with both SFB and clostridia. The number of immunoglobulin A (IgA)-producing cells in the lamina propria was more elevated in SFB-mice than in Clost-mice, not only in the ileum but also in the colon. The number of IgA-producing cells in the colons of Clost-mice was a little increased compared to that in GF mice. Taken together, SFB and clostridia promoted the development of both IEL and IgA-producing cells in the small intestine and that of only IEL in the large intestine, respectively, suggesting the occurrence of compartmentalization of the immunological responses to the indigenous bacteria between the small and large intestines. PMID- 10377133 TI - Dissemination of Listeria monocytogenes by infected phagocytes. AB - In vitro data suggest that blood-borne Listeria monocytogenes organisms enter the central nervous system (CNS) by direct invasion of endothelial cells or by cell to-cell spread from infected phagocytes to endothelial cells. However, a role for infected phagocytes in neuroinvasion and dissemination of L. monocytogenes in vivo has not been confirmed experimentally. Experiments described here tested whether L. monocytogenes-infected peripheral blood leukocytes (PBL) circulated in bacteremic mice and could establish organ infection in vivo. A mean of 30.5% of bacteria cultured from whole blood were PBL associated, and microscopy showed that 22.2% of monocytes and 1.6% of neutrophils were infected. PBL-associated bacteria spread to endothelial cells in vitro, indicating their potential for virulence in vivo. To test this possibility, mice were injected intravenously with infected PBL and CFU of bacteria in liver, spleen, and brain were quantified and compared with values for mice injected with broth-grown bacteria and in vitro infected macrophage cell lines. An inoculum of infected macrophage cell lines led to greater numbers of bacteria in the liver than the numbers produced by a similar inoculum of broth-grown bacteria. In contrast, brain infection was best established by infected PBL. Results of intraperitoneal injection of infected peritoneal cells compared with results of injection with infected J774A.1 cells suggested that unrestricted intracellular bacterial replication within J774A.1 cells contributed to excessive liver infection in those mice. These data show dissemination of intracellular L. monocytogenes and indicate that phagocyte facilitated invasion has a role in CNS infection in vivo. Heterogeneity with regard to bactericidal activity as well as to other phagocyte characteristics is a critical feature of this mechanism. PMID- 10377134 TI - Four clones of Borrelia burgdorferi sensu stricto cause invasive infection in humans. AB - Lyme disease begins at the site of a tick bite, producing a primary infection with spread of the organism to secondary sites occurring early in the course of infection. A major outer surface protein expressed by the spirochete early in infection is outer surface protein C (OspC). In Borrelia burgdorferi sensu stricto, OspC is highly variable. Based on sequence divergence, alleles of ospC can be divided into 21 major groups. To assess whether strain differences defined by ospC group are linked to invasiveness and pathogenicity, we compared the frequency distributions of major ospC groups from ticks, from the primary erythema migrans skin lesion, and from secondary sites, principally from blood and spinal fluid. The frequency distribution of ospC groups from ticks is significantly different from that from primary sites, which in turn is significantly different from that from secondary sites. The major groups A, B, I, and K had higher frequencies in the primary sites than in ticks and were the only groups found in secondary sites. We define three categories of major ospC groups: one that is common in ticks but very rarely if ever causes human disease, a second that causes only local infection at the tick bite site, and a third that causes systemic disease. The finding that all systemic B. burgdorferi sensu stricto infections are associated with four ospC groups has importance in the diagnosis, treatment, and prevention of Lyme disease. PMID- 10377135 TI - Analysis of a capsular polysaccharide biosynthesis locus of Bacteroides fragilis. AB - A major clinical manifestation of infection with Bacteroides fragilis is the formation of intra-abdominal abscesses, which are induced by the capsular polysaccharides of this organism. Transposon mutagenesis was used to locate genes involved in the synthesis of capsular polysaccharides. A 24,454-bp region was sequenced and found to contain a 15,379-bp locus (designated wcf) with 16 open reading frames (ORFs) encoding products similar to those encoded by genes of other bacterial polysaccharide biosynthesis loci. Four genes encode products that are similar to enzymes involved in nucleotide sugar biosynthesis. Seven genes encode products that are similar to sugar transferases. Two gene products are similar to O-acetyltransferases, and two products are probably involved in polysaccharide transport and polymerization. The product of one ORF, WcfH, is similar to a set of deacetylases of the NodB family. Deletion mutants demonstrated that the wcf locus is necessary for the synthesis of polysaccharide B, one of the two capsular polysaccharides of B. fragilis 9343. The virulence of the polysaccharide B-deficient mutant was comparable to that of the wild type in terms of its ability to induce abscesses in a rat model of intra-abdominal infection. PMID- 10377136 TI - Identification and characterization of TspA, a major CD4(+) T-cell- and B-cell stimulating Neisseria-specific antigen. AB - In search for novel T-cell immunogens involved in protection against invasive meningococcal disease, we screened fractionated proteins of Neisseria meningitidis (strain SD, B:15:P1.16) by using peripheral blood mononuclear cells (PBMCs) and specific T-cell lines obtained from normal individuals and patients convalescing from N. meningitidis infection. Proteins of iron-depleted meningococci produced higher PBMC proliferation indices than proteins of iron replete organisms, indicating that iron-regulated proteins are T-cell immunogens. Insoluble proteins of the iron-depleted cells, which produced better T-cell stimulation than soluble ones, were fractionated by using sodium dodecyl sulfate polyacrylamide gels and recovered as five fractions (F1 to F5) corresponding to decreasing molecular weight ranges. The proteins were purified (by elution and precipitation) or electroblotted onto nitrocellulose membranes (dissolved and precipitated) before use in further T-cell proliferation assays. One of the fractions (F1), containing high-molecular-mass proteins (>130 kDa), consistently showed the strongest T-cell proliferation responses in all of the T-cell lines examined. F1 proteins were subdivided into four smaller fractions (F1A to F1D) which were reexamined in T-cell proliferation assays, and F1C induced the strongest responses in patients' T-cell lines. Rabbit polyclonal antibodies to F1C components were used to screen a genomic expression library of N. meningitidis. Two major clones (C1 and C24) of recombinant meningococcal DNA were identified and fully sequenced. Sequence analysis showed that C24 (1,874 bp) consisted of a single open reading frame (ORF), which was included in clone C1 (2, 778 bp). The strong CD4(+) T-cell-stimulating effect of the polypeptide product of this ORF (named TspA) was confirmed, using a patient T-cell line. Immunogenicity for B cells was confirmed by showing that convalescent patients' serum antibodies recognized TspA on Western blots. Additional genetic sequence downstream of C24 was obtained from the meningococcal genomic sequence database (Sanger Centre), enabling the whole gene of 2,761 bp to be reconstructed. The DNA and deduced amino acid sequence data for tspA failed to show significant homology to any known gene, except for a corresponding (uncharacterized) gene in Neisseria gonorrhoeae genome sequences, suggesting that tspA is unique to the genus Neisseria. The DNA and deduced amino acid sequence of the second ORF of clone C1 showed significant homology to gloA, encoding glyoxalase I enzyme, of Salmonella typhimurium and Escherichia coli. Thus, we have identified a novel neisserial protein (TspA) which proved to be a strong CD4(+) T-cell- and B-cell-stimulating immunogen with potential as a possible vaccine candidate. PMID- 10377137 TI - Mouse beta-defensin 3 is an inducible antimicrobial peptide expressed in the epithelia of multiple organs. AB - One component of host defense at mucosal surfaces is epithelium-derived peptides with antimicrobial activity called defensins. We describe in this report the isolation and characterization of a murine homologue of human beta-defensin 2 (hBD-2) called mouse beta-defensin 3 (mBD-3). The predicted amino acid sequence shows the hallmark features of other known epithelial defensins, including the ordered array of six cysteine residues. Analysis of a genomic clone of mBD-3 revealed two exons separated by a 1.7-kb intron. The mBD-3 gene is localized at the proximal portion of chromosome 8, the site where genes for mouse alpha- and beta-defensins are found. Under basal condition, mBD-3 transcripts were detected at low levels in epithelial cells of surface organs, such as the intestine and lung. After instillation of Pseudomonas aeruginosa PAO1 into mouse airways, mBD-3 specific mRNA was upregulated significantly not only in large airways but also in the small bowel and liver. Recombinant mBD-3 peptide, produced from a baculovirus expression system, showed antimicrobial activity against P. aeruginosa PAO1 (MIC of 8 micrograms/ml) and Escherichia coli D31 (MIC of 16 micrograms/ml) in a salt dependent manner. This study demonstrates that a murine homologue of hBD-2 is present in the respiratory system and other mucosal surfaces. These similarities between murine and human host defense apparatus provide further impetus to evaluate the mouse as a model for studying the human innate host defense system. PMID- 10377138 TI - Role of the 85-kilobase plasmid and plasmid-encoded virulence-associated protein A in intracellular survival and virulence of Rhodococcus equi. AB - Rhodococcus equi is a facultative intracellular pathogen of macrophages and a cause of pneumonia in young horses (foals) and immunocompromised people. Isolates of R. equi from pneumonic foals typically contain large, 85- or 90-kb plasmids encoding a highly immunogenic virulence-associated protein (VapA). The objective of this study was to determine the role of the 85-kb plasmid and VapA in the intracellular survival and virulence of R. equi. Clinical isolates containing the plasmid and expressing VapA efficiently replicated within mouse macrophages in vitro, while plasmid-cured derivatives of these organisms did not multiply intracellularly. An isolate harboring the large plasmid also replicated in the tissues of experimentally infected mice, whereas its plasmid-cured derivative was rapidly cleared. All foals experimentally infected with a plasmid-containing clinical isolate developed severe bronchopneumonia, whereas the foals infected with its plasmid-cured derivative remained asymptomatic and free of visible lung lesions. By day 14 postinfection, lung bacterial burdens had increased considerably in foals challenged with the plasmid-containing clinical isolate. In contrast, bacteria could no longer be cultured from the lungs of foals challenged with the isogenic plasmid-cured derivative. A recombinant, plasmid-cured derivative expressing wild-type levels of VapA failed to replicate in macrophages and remained avirulent for both mice and foals. These results show that the 85-kb plasmid of R. equi is essential for intracellular replication within macrophages and for development of disease in the native host, the foal. However, expression of VapA alone is not sufficient to restore the virulence phenotype. PMID- 10377139 TI - Role of CD40 ligand in Mycobacterium avium infection. AB - Mycobacterium avium is a common opportunistic pathogen in immunocompromised patients such as those infected with human immunodeficiency virus. Although M. avium is an intracellular organism replicating predominantly in macrophages, disseminated M. avium infection is seen in AIDS patients with CD4(+) cell counts of <50 cells/microliters, suggesting a possible involvement of a T cell macrophage interaction for the elimination of M. avium. To determine whether CD40 CD40 ligand (CD40L) interactions play a role in M. avium infection, we studied the ability of CD40L to restrict M. avium replication in human monocyte-derived macrophages (MDM) in vitro. MDM were infected with M. avium and cocultured with CD40L-transfected 293 cells for 7 days. Intracellular growth of M. avium in these MDM was assessed by colony counting. CD40L-expressing cells inhibited growth of M. avium in MDM by 86.5% +/- 4.2% compared to MDM cultured with control cells. These findings were verified by assays using purified, soluble recombinant human CD40L (CD40LT). CD40LT (5 micrograms/ml) inhibited intracellular growth of M. avium by 76.9% +/- 18.0% compared to cells treated with medium alone. Inhibition by CD40LT was reduced by monoclonal antibodies (MAbs) against CD40 and CD40L. The inhibitory effect of CD40LT was not accompanied by enhancement of interleukin-12 (IL-12) production by M. avium-infected MDM, while CD40L-expressing cells stimulated IL-12 production by these cells. Treatment of M. avium-infected mice with MAb against murine CD40L resulted in recovery of larger numbers of organisms (0.8 to 1.0 log) from the spleens, livers, and lungs of these animals compared to infected mice which received normal immunoglobulin G. These results indicate that CD40-CD40L signaling may be an important step in host immune response against M. avium infection. PMID- 10377140 TI - The capsule supports survival but not traversal of Escherichia coli K1 across the blood-brain barrier. AB - The vast majority of cases of gram-negative meningitis in neonates are caused by K1-encapsulated Escherichia coli. The role of the K1 capsule in the pathogenesis of E. coli meningitis was examined with an in vivo model of experimental hematogenous E. coli K1 meningitis and an in vitro model of the blood-brain barrier. Bacteremia was induced in neonatal rats with the E. coli K1 strain C5 (O18:K1) or its K1(-) derivative, C5ME. Subsequently, blood and cerebrospinal fluid (CSF) were obtained for culture. Viable bacteria were recovered from the CSF of animals infected with E. coli K1 strains only; none of the animals infected with K1(-) strains had positive CSF cultures. However, despite the fact that their cultures were sterile, the presence of O18 E. coli was demonstrated immunocytochemically in the brains of animals infected with K1(-) strains and was seen by staining of CSF samples. In vitro, brain microvascular endothelial cells (BMEC) were incubated with K1(+) and K1(-) E. coli strains. The recovery of viable intracellular organisms of the K1(+) strain was significantly higher than that for the K1(-) strain (P = 0.0005). The recovery of viable intracellular K1( ) E. coli bacteria was increased by cycloheximide treatment of BMEC (P = 0.0059) but was not affected by nitric oxide synthase inhibitors or oxygen radical scavengers. We conclude that the K1 capsule is not necessary for the invasion of bacteria into brain endothelial cells but is responsible for helping to maintain bacterial viability during invasion of the blood-brain barrier. PMID- 10377142 TI - Early events in the pathogenesis of avian salmonellosis. AB - Salmonellae are gastrointestinal pathogens of man and animals. However, strains that are host-specific avian pathogens are often avirulent in mammals, and those which are nonspecific are commensal in poultry. The objective of this study was to determine whether host specificity was exhibited by bacterial abilities to invade epithelial cells or resist leukocyte killing. In this study, leukocytes isolated from humans and chickens were used to kill Salmonella in vitro. Both Salmonella pullorum, an avian-specific serotype, and Salmonella typhimurium, a broad-host-range serotype, were sensitive to killing by polymorphonuclear leukocytes isolated from both species. Both serotypes replicated in cells of the MQ-NCSU avian-macrophage cell line. In contrast, S. pullorum was noninvasive for cultured epithelial Henle 407, chick kidney, chick ovary, and budgerigar abdominal tumor cells. In the bird challenge, however, S. typhimurium rapidly caused inflammation of the intestinal mucosa, but S. pullorum preferentially targeted the bursa of Fabricius prior to eliciting intestinal inflammation. Salmonella serotypes which cause typhoid fever in mice have been shown to target the gut-associated lymphoid tissue. Observations from this study show that S. pullorum initiated a route of infection in chicks comparable to the route it takes in cases of enteric fever. PMID- 10377141 TI - Fatal granuloma necrosis without exacerbated mycobacterial growth in tumor necrosis factor receptor p55 gene-deficient mice intravenously infected with Mycobacterium avium. AB - The pathogenesis of mycobacterial infections is associated with the formation of granulomas in which both antibacterial protection and tissue damage take place concomitantly. We used murine Mycobacterium avium infection to compare the development of granulomatous lesions in intravenously infected tumor necrosis factor receptor p55 (TNFRp55) gene-deficient (p55(-/-)) mice to the development of granulomatous lesions in M. avium-infected syngeneic C57BL/6 (p55(+/+)) mice. Up to 5 weeks after infection with either the highly virulent M. avium strain TMC724 or the intermediately virulent M. avium strain SE01, bacterial counts in the liver, spleen, and lung of p55(-/-) mice were identical to or lower than those in infected p55(+/+) mice. However, the formation of mononuclear cell foci in the liver was delayed by approximately 2 to 3 weeks in p55(-/-) mice compared to the results obtained for infected p55(+/+) mice. Despite comparable bacterial loads, granulomas in p55(-/-) mice underwent progressive necrosis, causing damage to the surrounding liver tissue. The appearance of necrotizing granulomas was associated with the death of all infected p55(-/-) mice, regardless of the virulence of the M. avium strain used for infection. Granulomatous lesions in the liver contained three times as many CD3(+) cells in p55(-/-) mice yet appeared more diffuse than in p55(+/+) mice. Semiquantitative reverse transcription-PCR studies revealed that prior to mouse death, interleukin-12 (IL-12) and gamma interferon mRNA levels were up regulated in the livers of infected p55(-/-) mice, while mRNA levels for tumor necrosis factor, the inducible isoform of nitric oxide synthase (iNOS), and IL-10 were similar to those found in infected p55(+/+) mice. In response to persistent mycobacterial infection, the absence of TNFRp55 causes the disregulation of T-cell-macrophage interactions and results in fatal granuloma necrosis even when adequate antibacterial functions are maintained. PMID- 10377143 TI - Protective immunity of microsphere-based mucosal vaccines against lethal intranasal challenge with Streptococcus pneumoniae. AB - Mucosal vaccination of capsular polysaccharide (PS) of Streptococcus pneumoniae and subsequent creation of the first line of immunological defense in mucosa were examined. Mucosal as well as systemic antibody responses to PS were evoked by peroral or intranasal immunization of BALB/c mice with PS-cholera toxin B subunit (CTB) conjugates entrapped in the alginate microspheres (AM). The bacterial colonization at the lung mucosa was most profoundly inhibited (<95%) by intranasal immunization with the naked conjugate (PS-CTB). The mice vaccinated orally with encapsulated conjugate [AM(PS-CTB)] showed significant reduction on the level of pneumococcal bacteremia (<99%). Eighty percent of the mice perorally immunized with AM (PS-CTB) were protected from lethal intranasal challenge with S. pneumoniae, whereas more than 60% of the mice in the other control groups died of infection. Our novel approach may prove to be important in the development of a mucosal vaccine that will provide protection of mucosal surfaces of host. PMID- 10377144 TI - Obligatory role of gamma interferon for host survival in a murine model of infection with Burkholderia pseudomallei. AB - Burkholderia pseudomallei, the causative agent of melioidosis, is a gram-negative bacterium capable of causing either acute lethal sepsis or chronic but eventually fatal disease in infected individuals. However, despite the clinical importance of this infection in areas where it is endemic, there is essentially no information on the mechanisms of protective immunity to the bacterium. We describe here a murine model of either acute or chronic infection with B. pseudomallei in Taylor Outbred (TO) mice which mimics many features of the human pathology. Intraperitoneal infection of TO mice at doses of >10(6) CFU resulted in acute septic shock and death within 2 days. In contrast, at lower doses mice were able to clear the inoculum from the liver and spleen over a 3- to 4-week period, but persistence of the organism at other sites resulted in a chronic infection of between 2 and 16 months duration which was eventually lethal in all of the animals tested. Resistance to acute infection with B. pseudomallei was absolutely dependent upon the production of gamma interferon (IFN-gamma) in vivo. Administration of neutralizing monoclonal antibody against IFN-gamma lowered the 50% lethal dose from >5 x 10(5) to ca. 2 CFU and was associated with 8,500- and 4,400-fold increases in the bacterial burdens in the liver and spleen, respectively, together with extensive destruction of lymphoid architecture in the latter organ within 48 h. Neutralization of either tumor necrosis factor alpha or interleukin-12 but not granulocyte-macrophage colony-stimulating factor, also increased susceptibility to infection in vivo. Together, these results provide the first evidence of a host protective mechanism against B. pseudomallei. The rapid production of IFN-gamma within the first day of infection determines whether the infection proceeds to an acute lethal outcome or becomes chronic. PMID- 10377145 TI - Differential regulation of immune responses by highly and weakly virulent Cryptococcus neoformans isolates. AB - Early inflammatory responses, delayed-type hypersensitivity (DTH) responses, and cytokine profiles were studied in mice infected by the pulmonary route with either a highly virulent isolate (NU-2) or a weakly virulent isolate (184A) of Cryptococcus neoformans. After infection, NU-2 remained in the lungs and the capsule became more pronounced during the first 24 h, whereas 184A induced an immediate inflammatory reaction and was rapidly cleared from the lungs. Cryptococcal antigen (GXM) appeared in sera early after infection with NU-2 and increased over the entire observation period. There was no detectable GXM in sera from 184A-infected mice. Both C. neoformans isolates induced anticryptococcal cell-mediated immune responses, but the responses had different profiles. DTH in NU-2-infected mice appeared at day 15 after infection and waned by day 21, whereas DTH in 184A-infected mice was present by day 5 and continued to increase. T helper 1 (Th1) cytokines (interleukin 2 [IL-2] and gamma interferon) were made by spleen cells early after infection with either isolate. NU-2-infected mice lost their ability to produce these cytokines, but 184A-infected mice retained it. IL-4, a Th2 cytokine, was not detected in infected mice. The regulatory cytokine IL-10 was made by spleen cells early but not later after infection with the highly virulent isolate and was not produced by spleen cells from 184A infected mice. IL-10-deficient mice survived an NU-2 infection significantly longer than wild-type mice, suggesting that IL-10 is important in down-regulating the protective immune response. The induction of anergy appears to be responsible for the inability of NU-2-infected mice to control a C. neoformans infection. PMID- 10377146 TI - Resistance of virulent Mycobacterium avium to gamma interferon-mediated antimicrobial activity suggests additional signals for induction of mycobacteriostasis. AB - The cytokine gamma interferon (IFN-gamma) plays a major role in the control of Mycobacterium avium infections. We assessed whether the progressive growth of virulent strains of M. avium was associated with alterations in the production of this cytokine as evaluated by reverse transcription-PCR and detection of immunoreactive cytokine in the serum and in spleen homogenates. We found that IFN gamma was induced during infection by a virulent strain of M. avium to similar or even higher extents than the levels found during infections by a less virulent strain whose growth was controlled. IFN-gamma produced during infection by both mycobacterial strains was partly derived from T cells and led to activation of macrophages, namely, those that were infected. Concomitant with the development of the infection with the virulent strain of M. avium there was an extensive depletion of lymphocytes in the spleen. Thymectomy alone promoted the proliferation of the virulent, but not of the less virulent, strain of M. avium. Our data indicate that virulent strains of M. avium resist the antimicrobial mechanisms of IFN-gamma-activated macrophages and raise the possibility that a second, T-cell-dependent signal is required for the effective control of mycobacterial replication inside macrophages. PMID- 10377147 TI - Salmonella typhi flagella are potent inducers of proinflammatory cytokine secretion by human monocytes. AB - The cytokine production patterns of human peripheral blood mononuclear cells (PBMC) in response to Salmonella typhi flagella (STF) were examined in culture supernatants of PBMC stimulated with STF. Consistent with previous findings in volunteers vaccinated with aroC aroD deletion mutants of S. typhi, PBMC from volunteers immunized with the licensed live Ty21a S. typhi vaccine secreted gamma interferon following exposure to STF. Stimulation with STF induced rapid de novo synthesis of tumor necrosis factor alpha (TNF-alpha) and interleukin-1beta (IL 1beta), followed by IL-6 and IL-10. Trypsin treatment of STF abrogated their effects, while polymyxin B had no effect. Intracellular cytokine measurements of STF-stimulated PBMC revealed the existence of monocyte subpopulations that produce only TNF-alpha, IL-1beta or both cytokines. Moreover, STF markedly decreased the percentage of CD14(+) cells. These data demonstrate that STF are powerful monocyte activators which may have important implications for vaccine development and for understanding the pathogenesis of S. typhi infection. PMID- 10377148 TI - Pseudomonas aeruginosa gene products PilT and PilU are required for cytotoxicity in vitro and virulence in a mouse model of acute pneumonia. AB - Type IV pili of the opportunistic pathogen Pseudomonas aeruginosa mediate twitching motility and act as receptors for bacteriophage infection. They are also important bacterial adhesins, and nonpiliated mutants of P. aeruginosa have been shown to cause less epithelial cell damage in vitro and have decreased virulence in animal models. This finding raises the question as to whether the reduction in cytotoxicity and virulence of nonpiliated P. aeruginosa mutants are primarily due to defects in cell adhesion or loss of twitching motility, or both. This work describes the role of PilT and PilU, putative nucleotide-binding proteins involved in pili function, in mediating epithelial cell injury in vitro and virulence in vivo. Mutants of pilT and pilU retain surface pili but have lost twitching motility. In three different epithelial cell lines, pilT or pilU mutants of the strain PAK caused less cytotoxicity than the wild-type strain but more than isogenic, nonpiliated pilA or rpoN mutants. The pilT and pilU mutants also showed reduced association with these same epithelial cell lines compared both to the wild type, and surprisingly, to a pilA mutant. In a mouse model of acute pneumonia, the pilT and pilU mutants showed decreased colonization of the liver but not of the lung relative to the parental strain, though they exhibited no change in the ability to cause mortality. These results demonstrate that pilus function mediated by PilT and PilU is required for in vitro adherence and cytotoxicity toward epithelial cells and is important in virulence in vivo. PMID- 10377149 TI - Correlation of immunity in experimental syphilis with serum-mediated aggregation of Treponema pallidum rare outer membrane proteins. AB - We have previously shown by freeze-fracture electron microscopy that serum from infection-immune syphilitic rabbits aggregates the low-density membrane-spanning Treponema pallidum rare outer membrane proteins (TROMPs). The purpose of this study was to determine if a relationship could be demonstrated between acquired immunity in experimental rabbit syphilis, serum complement-dependent treponemicidal antibody, and antibody directed against TROMPs as measured by the aggregation of TROMP particles. Three groups of T. pallidum-infected rabbits were treated curatively with penicillin at 9 days, 30 days, and 6 months postinfection to generate various degrees of immunity to challenge reinfection. Sera from rabbits completely susceptible to localized and disseminated reinfection possessed a low titer of treponemicidal antibody (/=50% of a treponemal suspension) and showed a correspondingly low level of TROMP aggregation (16.5% of the total number of outer membrane particles counted) similar to normal serum controls (13. 4%); the number of particles within these aggregates never exceeded three. Sera from partially immune rabbits, which were susceptible to local reinfection but had no evidence of dissemination, showed an increase in the titer of treponemicidal antibody (1:16) compared to the completely susceptible group (C57BL/6 and LPL+/--->C57BL/6 mice, but not in LPL-/--->C57BL/6 mice, whereas myocardial cells expressed LPL in all groups. The mean aortic lesion area was reduced by 55% in LPL-/--->C57BL/6 mice compared with LPL+/+-->C57BL/6 mice and by 45% compared with LPL+/--->C57BL/6 mice, respectively. These data demonstrate in vivo that LPL expression by macrophages in the artery wall promotes foam cell formation and atherosclerosis. off PMID- 10377175 TI - Genetic dissection of lupus pathogenesis: a recipe for nephrophilic autoantibodies. AB - Sle1 and Sle3 are 2 loci that confer susceptibility to lupus nephritis in the NZM2410 strain of mice. Our previous work has shown that B6.NZMc1 mice, congenic for Sle1, exhibit loss of tolerance to chromatin but do not develop any pathogenic autoantibodies or disease. B6.NZMc7 mice, congenic for Sle3, exhibit low-grade polyclonal B- and T-cell activation, elevated CD4/CD8 ratios, and mildly penetrant glomerulonephritis. In contrast to these monocongenics, the present study reveals that B6.NZMc1|c7 mice, bicongenic for Sle1 and Sle3, exhibit splenomegaly, significantly expanded populations of activated B and CD4(+) T cells, and a robust, variegated IgG autoantibody response targeting multiple components of chromatin (including double-stranded DNA), intact glomeruli, and basement membrane matrix antigens. As one might predict, these mice, particularly the females, exhibit highly penetrant glomerulonephritis. These findings lend strong support to a two-step epistatic model for the formation of pathogenic, nephrophilic autoantibodies in lupus. Whereas loci such as Sle1 may serve to breach tolerance to chromatin, full-blown pathogenic maturation of the autoantibody response appears to require additional input from other loci (such as Sle3) and gender-based factors. PMID- 10377178 TI - Fundamental signals that regulate eosinophil homing to the gastrointestinal tract. AB - The histological identification of increased eosinophils in the gastrointestinal tract occurs in numerous clinical disorders; however, there is a limited understanding of the mechanisms regulating eosinophil trafficking into this mucosal surface. The results presented in this study characterize the processes regulating eosinophil homing into the gastrointestinal tract at baseline. Eosinophils were found to be present in the lamina propria of 19-day-old embryos and germ-free adult mice at concentrations comparable to those present in non germ-free adult mice. Furthermore, eosinophil gastrointestinal levels were not altered by increasing circulating eosinophils after pulmonary allergen challenge. Gastrointestinal eosinophil levels were partially reduced in mice deficient in recombinase activating gene-1 (RAG-1), IL-5, or the beta common chain (betac), but these reductions paralleled reductions in circulating eosinophils. In contrast, mice deficient in eotaxin had a marked reduction in gastrointestinal eosinophils but normal levels of eosinophils in the hematopoietic compartments. Furthermore, eotaxin was important for regulating eosinophil levels, even in the presence of high levels of IL-5. These investigations demonstrate eosinophil homing into the gastrointestinal tract during embryonic development occurring independently of viable intestinal flora. Furthermore, eotaxin is identified as the primary regulator of eosinophil gastrointestinal homing under homeostatic states, and may therefore have a fundamental role in innate immune responses. PMID- 10377177 TI - Short ragweed allergen induces eosinophilic lung disease in HLA-DQ transgenic mice. AB - The human leukocyte antigen (HLA) restriction of the IgE response to different allergens in humans has been a subject of numerous published studies. However, the role and contribution of specific HLA class II molecules in the pathogenesis of allergic airway inflammation are unknown and difficult to assess. HLA-DQ6 and HLA-DQ8 transgenic mice lacking endogenous mouse class II gene expression were actively immunized and later challenged intranasally with short ragweed (SRW) allergenic extract. The HLA-DQ transgenic mice developed pulmonary eosinophilia and lung tissue damage. We also found an increase in total protein (TP) level and IL-5 production in bronchoalveolar lavage (BAL) fluid and an increase in SRW specific Th2-type immunoglobulins (IgG1, IgG2b) and total serum IgE levels. Under similar treatment, DQ-negative full-sib control mice were normal. The allergic response could be significantly inhibited or abrogated in HLA-DQ mice by systemic treatment with anti-DQ mAb. The in vivo responses of HLA-DQ6 and HLA-DQ8 mice showed differences in terms of levels of eosinophilia, BAL protein, IL-5 concentration, and lung hyperreactivity to inhaled methacholine. These findings demonstrate the crucial role for specific HLA-DQ molecules in SRW-specific CD4(+) T-cell activation and resulting recruitment of eosinophils into the airways. PMID- 10377179 TI - Mitogen-activated protein kinase inhibits 1,25-dihydroxyvitamin D3-dependent signal transduction by phosphorylating human retinoid X receptor alpha. AB - Human retinoid X receptor alpha (hRXR alpha) is a member of the nuclear receptor family of transcriptional regulators. It regulates transcription through its association with several heterodimeric partners, including the vitamin D3 receptor (VDR). Signaling through the VDR is essential for normal calcium homeostasis and has been shown to inhibit the proliferation of cancer cells derived from a number of tissues. Here we show that phosphorylation of hRXR alpha in ras-transformed human keratinocytes through the activated Ras-Raf-mitogen activated protein kinase (Ras-Raf-MAP kinase) pathway results in attenuated transactivation by the VDR and resistance to the growth inhibitory action of 1,25 dihydroxyvitamin D3 [1,25(OH)2D3] and RXR-specific agonist LG1069 (4-[1-(5,6,7, 8 tetrahydro-3,5,5,8,8-pentamethyl-2-naphthalenyl) ethenyl]-benzoic acid). Phosphorylation of hRXR alpha occurs at serine 260, a consensus MAP kinase site. Inhibition of MAP kinase activity or point mutagenesis of serine 260 of hRXR alpha reverses the observed resistance to 1,25(OH)2D3 and LG1069. Thus, hRXR alpha is a downstream target of MAP kinase, and its phosphorylation may play an important role in malignant transformation. PMID- 10377180 TI - SLP-76 deficiency impairs signaling via the high-affinity IgE receptor in mast cells. AB - SLP-76 is an adapter protein expressed in T cells and myeloid cells that is a substrate for ZAP-70 and Syk. SLP-76-deficient mice exhibit a profound block in T cell development. We found that although SLP-76 is expressed in mouse mast cells, SLP-76(-/-) mice have normal numbers of mast cells in their skin and bronchi. SLP 76(-/-) mice are resistant to IgE-mediated passive anaphylaxis. SLP-76(-/-) mice sensitized with IgE anti-dinitrophenyl (DNP) and then challenged with DNP-HSA developed only mild and transient tachycardia, failed to increase their plasma histamine level, and all survived the antigen challenge. Bone marrow-derived mast cells (BMMCs) from SLP76(-/-) mice failed to release beta-hexosaminidase and to secrete IL-6 after FcepsilonRI cross-linking. Tyrosine phosphorylation of phospholipase C-gamma1 (but not of Syk) and calcium mobilization in response to IgE cross-linking were reduced in SLP-76-deficient BMMCs. These results suggest that SLP-76 plays an important role in FcepsilonRI-mediated signaling in mast cells. PMID- 10377181 TI - Natural killer cell-mediated lysis of autologous cells modified by gene therapy. AB - This study investigated the role of natural killer (NK) cells as effectors of an immune response against autologous cells modified by gene therapy. T lymphocytes were transduced with LXSN, a retroviral vector adopted for human gene therapy that carries the selectable marker gene neo, and the autologous NK response was evaluated. We found that (i) infection with LXSN makes cells susceptible to autologous NK cell-mediated lysis; (ii) expression of the neo gene is responsible for conferring susceptibility to lysis; (iii) lysis of neo-expressing cells is clonally distributed and mediated only by NK clones that exhibit human histocompatibility leukocyte antigen (HLA)-Bw4 specificity and bear KIR3DL1, a Bw4-specific NK inhibitory receptor; and (iv) the targets are cells from HLA Bw4(+) individuals. Finally, neo peptides anchoring to the Bw4 allele HLA-B27 interfered with KIR3DL1-mediated recognition of HLA-B27, i.e., they triggered NK lysis. Moreover, neo gene mutations preventing translation of two of the four potentially nonprotective peptides reduced KIR3DL1(+) NK clone-mediated autologous lysis. Thus, individuals expressing Bw4 alleles possess an NK repertoire with the potential to eliminate autologous cells modified by gene therapy. By demonstrating that NK cells can selectively detect the expression of heterologous genes, these observations provide a general model of the NK cell mediated control of viral infections. PMID- 10377183 TI - CD8(+) T cells mediate CD40-independent maturation of dendritic cells in vivo. AB - Induction of cytotoxic T lymphocyte (CTL) responses against minor histocompatibility antigens is dependent upon the presence of T cell help and requires the interaction of CD40 on dendritic cells (DCs) with CD40 ligand on activated T helper cells (Th). This study demonstrates that CD40 is neither involved in Th-dependent nor Th-independent antiviral CTL responses. Moreover, the data show that DC maturation occurs in vivo after viral infection in the absence of CD40 and Th. This maturation did not require viral infection of DCs but was mediated by peptide-specific CD8(+) T cells. Surprisingly, naive CD8(+) T cells were able to trigger DC maturation within 24 h after activation in vivo and in vitro. Moreover, peptide-activated CD8(+) T cells were able to induce maturation in trans, as DCs that failed to present the relevant antigen in vivo also underwent maturation. Upon isolation, the in vivo-stimulated DCs were able to convert a classically Th-dependent CTL response (anti-HY) into a Th independent response in vitro. Thus, antiviral CD8(+) T cells are sufficient for the maturation of DCs in the absence of CD40. PMID- 10377182 TI - Influence of polymorphism in the genes for the interleukin (IL)-1 receptor antagonist and IL-1beta on tuberculosis. AB - Several lines of evidence suggest that host genetic factors controlling the immune response influence infection by Mycobacterium tuberculosis. The proinflammatory cytokine interleukin (IL)-1beta and its antagonist, IL-1Ra (IL-1 receptor agonist), are strongly induced by M. tuberculosis and are encoded by polymorphic genes. The induction of both IL-1Ra mRNA and secreted protein by M. tuberculosis in IL-1Ra allele A2-positive (IL-1Ra A2(+)) healthy subjects was 1.9 fold higher than in IL-1Ra A2(-) subjects. The M. tuberculosis-induced expression of mRNA for IL-1beta was higher in subjects of the IL-1beta (+3953) A1(+) haplotype (P = 0.04). The molar ratio of IL-1Ra/IL-1beta induced by M. tuberculosis was markedly higher in IL-1Ra A2(+) individuals (P < 0.05), with minor overlap between the groups, reflecting linkage between the IL-1Ra A2 and IL 1beta (+3953) A2 alleles. In M. tuberculosis-stimulated peripheral blood mononuclear cells, the addition of IL-4 increased IL-1Ra secretion, whereas interferon gamma increased and IL-10 decreased IL-1beta production, indicative of a differential influence on the IL-1Ra/IL-1beta ratio by cytokines. In a study of 114 healthy purified protein derivative-reactive subjects and 89 patients with tuberculosis, the frequency of allelic variants at two positions (-511 and +3953) in the IL-1beta and IL-1Ra genes did not differ between the groups. However, the proinflammatory IL-1Ra A2(-)/IL-1beta (+3953) A1(+) haplotype was unevenly distributed, being more common in patients with tuberculous pleurisy (92%) in comparison with healthy M. tuberculosis-sensitized control subjects or patients with other disease forms (57%, P = 0.028 and 56%, P = 0. 024, respectively). Furthermore, the IL-1Ra A2(+) haplotype was associated with a reduced Mantoux response to purified protein derivative of M. tuberculosis: 60% of tuberculin nonreactive patients were of this type. Thus, the polymorphism at the IL-1 locus influences the cytokine response and may be a determinant of delayed-type hypersensitivity and disease expression in human tuberculosis. PMID- 10377184 TI - Immunoglobulin E-independent major histocompatibility complex-restricted T cell peptide epitope-induced late asthmatic reactions. AB - Intradermal administration of short overlapping peptides derived from chain 1 of the cat allergen Fel d 1 (FC1P) that did not cross-link IgE, elicited isolated late asthmatic reactions with no visible early or late cutaneous response in 9/40 cat-allergic asthmatics. Four of the nine were human histocompatibility leukocyte antigen DR13-positive, as compared with only 1/31 nonreactors. The other five reactors expressed either DR1 or DR4. To confirm major histocompatibility complex restriction, fibroblast cell lines transfected with HLA-DR molecules were used to present FC1Ps to cat allergen-specific T cell lines derived from subjects before peptide injection. FC1P3 (peptide 28-44 of Fel d 1 chain 1) was recognized in the context of DR13 alleles (DRB1*1301, 1302) and induced specific T cell proliferation and IL-5 production. T cells from a DR1(+) responder proliferated and produced IL-5 in the presence of FC1P3 and DR1 (DRB1*0101) fibroblast cell lines, whereas T cells from a DR4(+) subject recognized FC1P2 (peptide 22-37) when presented by DRB1*0405. We conclude that short, allergen-derived peptides can directly initiate a major histocompatibility complex-restricted, T cell dependent late asthmatic reaction, without the requirement for an early IgE/mast cell-dependent response, in sensitized asthmatic subjects. PMID- 10377185 TI - Cure of progressive murine leishmaniasis: interleukin 4 dominance is abolished by transient CD4(+) T cell depletion and T helper cell type 1-selective cytokine therapy. AB - Progressive infection with Leishmania major in susceptible BALB/c mice is mediated by interleukin (IL)-4-producing T helper cell type 2 (Th2) CD4(+) T cells that, once established, become resistant to Th1-deviating therapies with recombinant (r)IL-12 and/or neutralizing anti-IL-4 antibodies. We sought to restore protective immunity in advanced leishmaniasis by depletion of Th2-biased CD4(+) populations and by cytokine-directed reconstitution of Th1 cellular responses during lymphocyte recovery. Treatment with cytolytic GK1.5 anti-CD4 mAb alone did not reverse disease in 3 wk-infected BALB/c mice, but GK1.5 combined with anti-IL-4 antibody and intralesional rIL-12 cured cutaneous lesions in 80% of mice and established a Th1-polarized cytokine response to L. major antigen protective against reinfection. The curative effects of GK1.5 were not replaced by cytotoxic anti-CD8 monoclonal antibody 2.43 or nondepleting anti-CD4 mAb YTS177, confirming that depletion of CD4(+) cells was specific and essential for therapeutic effect. Finally, combined CD4(+) depletion and IL-4 neutralization were curative, indicating that neither increased parasite burden nor altered accessory cell function independently biased towards Th2 reconstitution in advanced leishmaniasis. Advanced leishmaniasis can be cured by T cell depletion and cytokine-directed recovery of Th1 cellular responses, suggesting novel interventions for other immune-mediated diseases and identifying distinct roles for CD4(+) T cell and non-T cell in the maintenance of Th2 and Th1 phenotypes. PMID- 10377186 TI - An invariant T cell receptor alpha chain defines a novel TAP-independent major histocompatibility complex class Ib-restricted alpha/beta T cell subpopulation in mammals. AB - We describe here a new subset of T cells, found in humans, mice, and cattle. These cells bear a canonical T cell receptor (TCR) alpha chain containing hAV7S2 and AJ33 in humans and the homologous AV19-AJ33 in mice and cattle with a CDR3 of constant length. These T cells are CD4(-)CD8(-) double-negative (DN) T cells in the three species and also CD8alphaalpha in humans. In humans, their frequency was approximately 1/10 in DN, 1/50 in CD8alpha+, and 1/6,000 in CD4(+) lymphocytes, and they display an activated/memory phenotype (CD45RAloCD45RO+). They preferentially use hBV2S1 and hBV13 segments and have an oligoclonal Vbeta repertoire suggesting peripheral expansions. These cells were present in major histocompatibility complex (MHC) class II- and transporter associated with antigen processing (TAP)-deficient humans and mice and also in classical MHC class I- and CD1-deficient mice but were absent from beta2-microglobulin deficient mice, indicating their probable selection by a nonclassical MHC class Ib molecule distinct from CD1. The conservation between mammalian species, the abundance, and the unique selection pattern suggest an important role for cells using this novel canonical TCR alpha chain. PMID- 10377187 TI - Lipoxin (LX)A4 and aspirin-triggered 15-epi-LXA4 inhibit tumor necrosis factor 1alpha-initiated neutrophil responses and trafficking: regulators of a cytokine chemokine axis. AB - The impact of lipoxin A4 (LXA4) and aspirin-triggered lipoxins (ATLs) was investigated in tumor necrosis factor (TNF)-alpha-initiated neutrophil (polymorphonuclear leukocyte) responses in vitro and in vivo using metabolically stable LX analogues. At concentrations as low as 1-10 nM, the LXA4 and ATL analogues each inhibited TNF-alpha-stimulated superoxide anion generation and IL 1beta release by human polymorphonuclear leukocytes. These LXA4-ATL actions were time and concentration dependent and proved selective for TNF-alpha, as these responses were not altered with either GM-CSF- or zymosan-stimulated cells. TNF alpha-induced IL-1beta gene expression was also regulated by both anti-LXA4 receptor antibodies and LXA4-ATL analogues. In murine air pouches, 15R/S-methyl LXA4 dramatically inhibited TNF-alpha-stimulated leukocyte trafficking, as well as the appearance of both macrophage inflammatory peptide 2 and IL-1beta, while concomitantly stimulating IL-4 in pouch exudates. Together, these results indicate that both LXA4 and ATL regulate TNF-alpha-directed neutrophil actions in vitro and in vivo and stimulate IL-4 in exudates, playing a pivotal role in immune responses. PMID- 10377189 TI - Interleukin 13 is secreted by and stimulates the growth of Hodgkin and Reed Sternberg cells. AB - Gene expression patterns can provide vital clues to the pathogenesis of neoplastic diseases. We investigated the expression of 950 genes in Hodgkin's disease (HD) by analyzing differential mRNA expression using microarrays. In two independent microarray experiments, the HD-derived cell lines L428 and KMH2 were compared with an Epstein-Barr virus (EBV)-immortalized lymphoblastoid B cell line, LCL-GK. Interleukin (IL)-13 and IL-5 were found to be highly expressed in the HD-derived cell lines. Examination of IL-13 and IL-5 expression by Northern blot analysis and enzyme-linked immunosorbent assay confirmed these results and revealed the expression of IL-13 in a third HD-derived cell line, HDLM2. Control LCL and EBV-negative non-Hodgkin lymphoma-derived cell lines did not express IL 13. In situ hybridization of lymph node tissue from HD patients showed that elevated levels of IL-13 were specifically expressed by Hodgkin/Reed-Sternberg (H/RS) tumor cells. Treatment of a HD-derived cell line with a neutralizing antibody to IL-13 resulted in a dose-dependent inhibition of H/RS cell proliferation. These data suggest that H/RS cells produce IL-13 and that IL-13 plays an important role in the stimulation of H/RS cell growth, possibly by an autocrine mechanism. Modulation of the IL-13 signaling pathway may be a logical objective for future therapeutic strategies. PMID- 10377188 TI - Chlamydia inhibits interferon gamma-inducible major histocompatibility complex class II expression by degradation of upstream stimulatory factor 1. AB - We report that chlamydiae, which are obligate intracellular bacterial pathogens, can inhibit interferon (IFN)-gamma-inducible major histocompatibility complex (MHC) class II expression. However, the IFN-gamma-induced IFN regulatory factor-1 (IRF-1) and intercellular adhesion molecule 1 (ICAM-1) expression is not affected, suggesting that chlamydia may selectively target the IFN-gamma signaling pathways required for MHC class II expression. Chlamydial inhibition of MHC class II expression is correlated with degradation of upstream stimulatory factor (USF)-1, a constitutively and ubiquitously expressed transcription factor required for IFN-gamma induction of class II transactivator (CIITA) but not of IRF-1 and ICAM-1. CIITA is an obligate mediator of IFN-gamma-inducible MHC class II expression. Thus, diminished CIITA expression as a result of USF-1 degradation may account for the suppression of the IFN-gamma-inducible MHC class II in chlamydia-infected cells. These results reveal a novel immune evasion strategy used by the intracellular bacterial pathogen chlamydia that improves our understanding of the molecular basis of pathogenesis. PMID- 10377190 TI - Structure and expression of an adhesive protein-like molecule of mosquito invasive-stage malarial parasite. AB - Invasion of the malarial parasite into a vector mosquito begins when the motile ookinete transverses the gut epithelium. Adhesive proteins that may mediate this invasive process have not been identified to date. We found that a molecule with an adhesive protein-like structure was expressed in the ookinete of Plasmodium berghei. This protein is structurally homologous to circumsporozoite protein and thrombospondin-related adhesive protein (TRAP)-related protein, CTRP, of Plasmodium falciparum. We named it P. berghei CTRP (PbCTRP) and report here its structure and manner of expression. PbCTRP has six integrin I region-like domains and seven thrombospondin-like domains in its putative extracellular region. This structure is similar to that of CTRP and TRAPs of malaria sporozoite. The putative transmembrane and cytoplasmic regions of PbCTRP, CTRP, and TRAP also have conserved amino acid sequences. PbCTRP is produced at least 10 h after fertilization when zygotes begin transformation to ookinetes. In the mature ookinete, PbCTRP is located mainly in the anterior cytoplasm. The staining pattern was also similar to TRAP in the sporozoite. We suggest that PbCTRP may play a role in ookinete invasive motility and belongs to a protein family together with TRAP and other structurally related proteins of apicomplexan parasites. PMID- 10377191 TI - Human herpesvirus 6 (HHV-6) causes severe thymocyte depletion in SCID-hu Thy/Liv mice. AB - Human herpesvirus 6 (HHV-6) is a potentially immunosuppressive agent that may act as a cofactor in the progression of AIDS. Here, we describe the first small animal model of HHV-6 infection. HHV-6 subgroup A, strain GS, efficiently infected the human thymic tissue implanted in SCID-hu Thy/Liv mice, leading to the destruction of the graft. Viral DNA was detected in Thy/Liv implants by quantitative polymerase chain reaction (PCR) as early as 4 d after inoculation and peaked at day 14. The productive nature of the infection was confirmed by electron microscopy and immunohistochemical staining. Atypical thymocytes with prominent nuclear inclusions were detected by histopathology. HHV-6 replication was associated with severe, progressive thymocyte depletion involving all major cellular subsets. However, intrathymic T progenitor cells (ITTPs) appeared to be more severely depleted than the other subpopulations, and a preferred tropism of HHV-6 for ITTPs was demonstrated by quantitative PCR on purified thymocyte subsets. These findings suggest that thymocyte depletion by HHV-6 may be due to infection and destruction of these immature T cell precursors. Similar results were obtained with strain PL-1, a primary isolate belonging to subgroup B. The severity of the lesions observed in this animal model underscores the possibility that HHV-6 may indeed be immunosuppressive in humans. PMID- 10377192 TI - Mannose receptor and its putative ligands in normal murine lymphoid and nonlymphoid organs: In situ expression of mannose receptor by selected macrophages, endothelial cells, perivascular microglia, and mesangial cells, but not dendritic cells. AB - The mannose receptor (MR) has established roles in macrophage (Mphi) phagocytosis of microorganisms and endocytic clearance of host-derived glycoproteins, and has recently been implicated in antigen capture by dendritic cells (DCs) in vitro. MR is the founder member of a family of homologous proteins, and its recognition properties differ according to its tissue of origin. Given this heterogeneity and our recent discovery of a soluble form of MR in mouse serum, we studied the sites of synthesis of MR mRNA and expression of MR protein in normal mouse tissues. We demonstrate that synthesis and expression occur at identical sites, and that mature Mphi and endothelium are heterogeneous with respect to MR expression, additionally describing MR on perivascular microglia and glomerular mesangial cells. However, MR was not detected on DCs in situ, or on marginal zone or subcapsular sinus Mphi, both of which have MR-like binding activities. We also compared expression of MR to the binding of a recombinant probe containing the cysteine-rich domain of MR. We show that MR and its putative ligand(s) are expressed at nonoverlapping sites within lymphoid organs, consistent with a transfer function for soluble MR. Therefore, in addition to endocytic and phagocytic roles, MR may play an important role in antigen recognition and transport within lymphoid organs. PMID- 10377193 TI - Susceptibility of mice deficient in CD1D or TAP1 to infection with Mycobacterium tuberculosis. AB - Cellular immunity against Mycobacterium tuberculosis controls infection in the majority of infected humans. Studies in mice have delineated an important role for CD4(+) T cells and cytokines including interferon gamma and tumor necrosis factor alpha in the response to infection with mycobacteria. Recently, the identification of CD8(+) CD1-restricted T cells that kill M. tuberculosis organisms via granulysin and the rapid death after infection of beta2 microglobulin deficient mice in humans has drawn attention to a critical role for CD8(+) T cells. The nature of mycobacterial-specific CD8(+) T cells has been an enigma because few have been identified in any species. Here, we delineate the contribution of class I MHC-restricted T cells in the defense against tuberculosis as transporter associated with antigen processing (TAP)1-deficient mice died rapidly, bore a greater bacterial burden, and had more severe tissue pathology than control mice. In contrast, CD1D-/- mice were not significantly different in their susceptibility to infection than control mice. This data demonstrates a critical role for TAP-dependent peptide antigen presentation and provides further evidence that class I MHC-restricted CD8(+) T cells, the major T cell subset activated by this antigen processing pathway, play an essential role in immunity to tuberculosis. PMID- 10377194 TI - Interleukin 12-dependent interferon gamma production by CD8alpha+ lymphoid dendritic cells. AB - We investigated the role of antigen-presenting cells in early interferon (IFN) gamma production in normal and recombinase activating gene 2-deficient (Rag-2(-/ )) mice in response to Listeria monocytogenes (LM) infection and interleukin (IL) 12 administration. Levels of serum IFN-gamma in Rag-2(-/-) mice were comparable to those of normal mice upon either LM infection or IL-12 injection. Depletion of natural killer (NK) cells by administration of anti-asialoGM1 antibodies had little effect on IFN-gamma levels in the sera of Rag-2(-/-) mice after LM infection or IL-12 injection. Incubation of splenocytes from NK cell-depleted Rag 2(-/-) mice with LM resulted in the production of IFN-gamma that was completely blocked by addition of anti-IL-12 antibodies. Both dendritic cells (DCs) and monocytes purified from splenocytes were capable of producing IFN-gamma when cultured in the presence of IL-12. Intracellular immunofluorescence analysis confirmed the IFN-gamma production from DCs. It was further shown that IFN-gamma was produced predominantly by CD8alpha+ lymphoid DCs rather than CD8alpha- myeloid DCs. Collectively, our data indicated that DCs are potent in producing IFN-gamma in response to IL-12 produced by bacterial infection and play an important role in innate immunity and subsequent T helper cell type 1 development in vivo. PMID- 10377195 TI - Dominant myelopoietic effector functions mediated by chemokine receptor CCR1. AB - Macrophage inflammatory protein (MIP)-1alpha, a CC chemokine, enhances proliferation of mature subsets of myeloid progenitor cells (MPCs), suppresses proliferation of immature MPCs, and mobilizes mature and immature MPCs to the blood. MIP-1alpha binds at least three chemokine receptors. To determine if CCR1 was dominantly mediating the above activities of MIP-1alpha, CCR1-deficient (-/-) mice, produced by targeted gene disruption, were used. MIP-1alpha enhanced colony formation of marrow granulocyte/macrophage colony-forming units (CFU-GM), responsive to stimulation by granulocyte/macrophage colony-stimulating factor (GM CSF), and CFU-M, responsive to stimulation by M-CSF, from littermate control CCR1(+/+) but not CCR1(-/-) mice. Moreover, MIP-1alpha did not mobilize MPCs to the blood or synergize with G-CSF in this effect in CCR1(-/-) mice. However, CCR1(-/-) mice were increased in sensitivity to MPC mobilizing effects of G-CSF. Multi-growth factor-stimulated MPCs in CCR1(-/-) and CCR1(+/+) marrow were equally sensitive to inhibition by MIP-1alpha. These results implicate CCR1 as a dominant receptor for MIP-1alpha enhancement of proliferation of lineage committed MPCs and for mobilization of MPCs to the blood. CCR1 is not a dominant receptor for MIP-1alpha suppression of MPC proliferation, but it does negatively impact G-CSF-induced MPC mobilization. PMID- 10377197 TI - Exhaled nitric oxide in COPD: glancing through a smoke screen. PMID- 10377196 TI - The Kaposi's sarcoma-related herpesvirus (KSHV)-encoded chemokine vMIP-I is a specific agonist for the CC chemokine receptor (CCR)8. AB - The Kaposi's sarcoma-related herpesvirus (KSHV), also designated human herpesvirus 8, is the presumed etiologic agent of Kaposi's sarcoma and certain lymphomas. Although KSHV encodes several chemokine homologues (viral macrophage inflammatory protein [vMIP]-I, -II, and -III), only vMIP-II has been functionally characterized. We report here that vMIP-I is a specific agonist for the CC chemokine receptor (CCR)8 that is preferentially expressed on Th2 T cells. Y3 cells transfected with CCR8 produced a calcium flux in response to vMIP-I and responded vigorously in in vitro chemotaxis assays. In competition binding experiments, the interaction of vMIP-I with CCR8 was shown to be specific and of high affinity. In contrast to its agonist activity at CCR8, vMIP-I did not interact with CCR5 or any of 11 other receptors examined. Furthermore, vMIP-I was unable to inhibit CCR5-mediated HIV infection. These findings suggest that expression of vMIP-I by KSHV may influence the Th1/Th2 balance of the host immune response. PMID- 10377198 TI - Tuberculosis in AIDS: past or new problems? PMID- 10377199 TI - Increased exhaled nitric oxide in patients with stable chronic obstructive pulmonary disease. AB - BACKGROUND: Nitric oxide (NO) plays an important role as an inflammatory mediator in the airways. Since chronic obstructive pulmonary disease (COPD) is characterised by airway inflammation, a study was undertaken to determine NO levels in the exhaled air of patients with COPD. METHODS: Two groups of patients with clinically stable COPD were studied, 10 current smokers and 10 ex-smokers. Two control groups of healthy subjects consisting of 10 current smokers and 20 non-smokers were also studied. Exhaled NO levels were measured by the collection bag technique and NO chemiluminescence analyser. RESULTS: Mean (SE) levels of exhaled NO in ex-smokers and current smokers with COPD (25.7 (3.0) ppb and 10.2 (1.4) ppb, respectively) were significantly higher than in non-smoker and current smoker control subjects (9.4 (0.8) ppb and 4.6 (0.4) ppb, respectively). In current smokers with COPD exhaled levels of NO were significantly lower than in ex-smokers. In this latter group of patients there was a significant negative correlation between smoking history (pack years) and levels of exhaled NO (r = 0.8, p = 0.002). A positive correlation was seen between forced expiratory volume in one second (FEV1) and levels of exhaled NO (r = 0.65, p = 0.001) in patients with COPD. CONCLUSIONS: These data show that exhaled NO is increased in patients with stable COPD, both current and ex-smokers, compared with healthy control subjects. PMID- 10377200 TI - Markers of nitric oxide metabolism in sputum and exhaled air are not increased in chronic obstructive pulmonary disease. AB - BACKGROUND: Nitric oxide (NO) is involved in inflammation and host defence of the lung. It has been found in increased concentrations in the airways in asthmatic subjects but its levels in patients with chronic obstructive pulmonary disease (COPD) have not been investigated. A study was undertaken to determine whether markers of NO metabolism (NO in exhaled air, iNOS expression in sputum cells, and nitrite + nitrate (NO2-/NO3-) in sputum supernatant) are increased in subjects with COPD, and whether they correlate with inflammatory indices in induced sputum. The associations of these markers with smoking were also assessed. METHODS: Sixteen subjects with COPD (median age 66 years, median forced expiratory volume in one second (FEV1) 63% predicted, eight current smokers) and 16 healthy subjects (median age 63 years, median FEV1 113% predicted, eight current smokers) participated in the study. NO was measured during tidal breathing and sputum was induced by inhalation of hypertonic saline. RESULTS: No differences were observed between subjects with COPD and healthy controls in exhaled NO excretion rate (median 5.15 and 6.25 nmol/min), sputum macrophage iNOS expression (14% and 12%), and sputum supernatant NO2-/NO3- (46 and 73 microM). NO in exhaled air correlated with the percentage of sputum eosinophils in patients with COPD (rho = 0.65, p = 0.009) but not in healthy individuals. Exhaled NO and supernatant NO2-/NO3- levels were lower in healthy smokers than in healthy non/ex smokers. CONCLUSIONS: Our findings indicate that NO metabolism is not increased in patients with stable COPD. The close association between exhaled NO levels and sputum eosinophils suggests a role for NO in airway inflammation in COPD. Studies performed during exacerbations may clarify this role. PMID- 10377201 TI - Usefulness of the Medical Research Council (MRC) dyspnoea scale as a measure of disability in patients with chronic obstructive pulmonary disease. AB - BACKGROUND: Methods of classifying chronic obstructive pulmonary disease (COPD) depend largely upon spirometric measurements but disability is only weakly related to measurements of lung function. With the increased use of pulmonary rehabilitation, a need has been identified for a simple and standardised method of categorising disability in COPD. This study examined the validity of the Medical Research Council (MRC) dyspnoea scale for this purpose. METHODS: One hundred patients with COPD were recruited from an outpatient pulmonary rehabilitation programme. Assessments included the MRC dyspnoea scale, spirometric tests, blood gas tensions, a shuttle walking test, and Borg scores for perceived breathlessness before and after exercise. Health status was assessed using the St George's Respiratory Questionnaire (SGRQ) and Chronic Respiratory Questionnaire (CRQ). The Nottingham Extended Activities of Daily Living (EADL) score and Hospital Anxiety and Depression (HAD) score were also measured. RESULTS: Of the patients studied, 32 were classified as having MRC grade 3 dyspnoea, 34 MRC grade 4 dyspnoea, and 34 MRC grade 5 dyspnoea. Patients with MRC grades 1 and 2 dyspnoea were not included in the study. There was a significant association between MRC grade and shuttle distance, SGRQ and CRQ scores, mood state and EADL. Forced expiratory volume in one second (FEV1) was not associated with MRC grade. Multiple logistic regression showed that the determinants of disability appeared to vary with the level of disability. Between MRC grades 3 and 4 the significant covariates were exercise performance, SGRQ and depression score, whilst between grades 4 and 5 exercise performance and age were the major determinants. CONCLUSIONS: The MRC dyspnoea scale is a simple and valid method of categorising patients with COPD in terms of their disability that could be used to complement FEV1 in the classification of COPD severity. PMID- 10377202 TI - Asymptomatic bronchial hyperresponsiveness to exercise in childhood and the development of asthma related symptoms in young adulthood: the Odense Schoolchild Study. AB - BACKGROUND: Exercise testing may be of value in identifying a group of children at high risk of subsequently developing respiratory symptoms. As few longitudinal studies have investigated this issue, the bronchial hyperresponsiveness to exercise in asymptomatic children was evaluated as a risk factor for developing asthma related symptoms in young adulthood. METHODS: A community based sample of 1369 schoolchildren, first investigated in 1985 at a mean age of 9.7 years, was followed up after a mean of 10.5 years. Nine hundred and twenty children (67%) were asymptomatic in childhood and 777 (84.9%) of these were re-investigated at follow up. At the first examination a maximum progressive exercise test on a bicycle ergometer was used to induce airway narrowing. The forced expiratory volume in one second (FEV1) after exercise was considered abnormal if the percentage fall in FEV1 was more than 5% of the highest fall in the reference subjects characterised by having no previous history of asthma or asthma related symptoms. The threshold for a positive test was 8.6% of pre-exercise FEV1. RESULTS: One hundred and three subjects (13%) had wheeze within the last year at follow up and, of these, nine (9%) had been hyperresponsive to exercise in 1985. One hundred and seventy subjects (22%) had non-infectious cough within the previous year, 11 of whom (6%) had been hyperresponsive to exercise in 1985. Multiple regression analysis showed that subjects with hyperresponsiveness to exercise had an increased risk of developing wheeze compared with subjects with a normal response to exercise when the fall in FEV1 after exercise was included as a variable (threshold odds ratio (OR) 2.3 (95% CI 1.1 to 5.5)). The trend was not significant when exercise induced bronchospasm was included as a continuous variable (OR 1.02 (95% CI 0.97 to 1.06)). CONCLUSIONS: Asymptomatic children who are hyperresponsive to exercise are at increased risk of developing new symptoms related to wheezing but the predictive value of exercise testing for individuals is low. PMID- 10377203 TI - Immunohistochemical localisation of the matrix metalloproteinases MMP-3 and MMP-9 within the airways in asthma. AB - BACKGROUND: The matrix metalloproteinase (MMP) enzymes MMP-3 and MMP-9 have relevance to the chronic structural airway changes in asthma. These proteinases can be generated by structural and inflammatory cells, and have the ability to degrade proteoglycans and thus potentially enhance airway fibrosis and smooth muscle proliferation through their ability to release and activate latent matrix bound growth factors. METHODS: Immunostaining for MMP-3 and MMP-9 as well as for mast cells, eosinophils, and neutrophils was undertaken in acetone fixed and glycolmethacrylate embedded endobronchial biopsy specimens obtained by fibreoptic bronchoscopy under local anaesthesia. The findings from 17 asthmatic subjects (nine with mild to moderate non-steroid treated asthma and eight with chronic persistent steroid-dependent asthma) were compared with those from eight healthy controls. The cell associated MMP immunoreactivity was co-localised to mast cells, eosinophils, or neutrophils and represented as cells/mm2, based on the area of the biopsy specimen. Extracellular matrix immunoreactivity was assessed by an image analysis system and visually with ranking and the two approaches were compared. RESULTS: The biopsy specimens from asthmatic subjects contained significantly more eosinophils (p<0. 001) than those from the non-asthmatic subjects. Both MMP-9 and MMP-3 immunoreactivity could be identified in endobronchial biopsy specimens. Gelatinase B (MMP-9) immunoreactivity was prominent within the extracellular matrix as well as exhibiting distinct cell immunoreactivity which predominantly co-localised to neutrophils. Stromelysin (MMP-3) was co-localised to mast cells, eosinophils, and neutrophils as well as being present in the epithelium, the lamina reticularis and, to a lesser extent, the extracellular matrix. There was no significant difference in the extent of matrix immunoreactivity for either MMP-3 or MMP-9 between healthy controls or subjects with mild or severe asthma. CONCLUSION: Although immunostaining cannot distinguish between active and inactive forms of MMPs, the presence of MMP-3 and MMP-9 within endobronchial biopsy specimens, the co-localisation to inflammatory cells of relevance to asthma (mast cells and eosinophils), and the identification of matrix binding, indicative of MMP-matrix interactions, points to the potential for disease related changes in MMP release that influence airway remodelling in asthma. PMID- 10377204 TI - Association of air pollution with daily GP consultations for asthma and other lower respiratory conditions in London. AB - BACKGROUND: Very few published studies have looked at the effects of air pollution on health in the primary care setting. As part of a large study to examine the association between air pollution and a number of health outcomes, the relationship between daily GP consultations for asthma and other lower respiratory diseases (LRD) and air pollution in London was investigated. METHODS: Time-series analysis of daily numbers of GP consultations controlling for time trends, seasonal factors, day of week cycles, influenza, weather, pollen levels, and serial correlation was performed. Consultation data were available from between 268 718 and 295 740 registered patients from 45-47 London practices contributing to the General Practice Research Database during 1992-4. RESULTS: Positive associations, weakly significant and consistent across lags, were observed between asthma consultations and nitrogen dioxide (NO2) and carbon monoxide (CO) in children and particulate matter of less than 10 microm in diameter (PM10) in adults, and between other LRD consultations and sulphur dioxide (SO2) in children. A consistently negative association with ozone in children was observed in both disease categories. The effect estimates of most pollutants were much larger when analysed separately by season, particularly in the children: percentage change in asthma consultations during the warm season (April-September) for a 10-90th percentile increase in 24 hour NO2 lagged by one day = 13.2% (95% CI 5.6 to 21.3), with CO = 11.4% (95% CI 3.3 to 20.0), and with SO2 = 9.0% (95% CI 2.2 to 16.2). In adults the only association consistent over different lag periods was with PM10 = 9.2% (3.7 to 15.1). The associations of pollution and consultations for LRD were increased mainly in the winter months: percentage change in consultations by children in winter with NO2 = 7.2% (95% CI 2.8 to 11.6), CO = 6.2% (95% CI 2.3 to 10.2), and SO2 = 5.8% (95% CI 1.6 to 10.2). CONCLUSIONS: There are associations between air pollution and daily consultations for asthma and other lower respiratory disease in London. The most significant associations were observed in children and the most important pollutants were NO2, CO, and SO2. In adults the only consistent association was with PM10. PMID- 10377205 TI - Prevalence of atopy, asthma symptoms and diagnosis, and the management of asthma: comparison of an affluent and a non-affluent country. AB - BACKGROUND: The prevalence of childhood asthma and of atopy varies widely between countries. However, few studies have compared the pattern of diagnosis and management of asthma, or the role of atopy in predisposing to asthma between a less affluent country and a more affluent country. The aim of this study was to compare the prevalence of symptoms, diagnosis, and management of asthma, and the prevalence of atopy as measured by skin prick tests in Nigeria and Australia using a standardised methodology. METHODS: Respiratory history was collected using a validated questionnaire administered to parents, and atopy was measured with skin prick tests in 654 Australian and 566 Nigerian children aged 8-11 years (70% consent rate in Australia, 60% in Nigeria). RESULTS: Wheeze and persistent cough were less prevalent in Nigeria (10.2% and 5.1%, respectively) than in Australia (21.9% and 9.6%, respectively), caused less morbidity, and were less likely to be labelled or treated as asthma than in Australia. There was no significant difference in the overall prevalence of atopy between the two countries (Australia 32. 5%, Nigeria 28.2%). Atopy was a strong risk for wheeze in both countries (odds ratio (OR) 3.4 (95% CI 2.3 to 5.1) in Australia, 1.8 (95% CI 1.0 to 3.3) in Nigeria), especially atopy to house dust mites (OR 3.1 (95% CI 2.1 to 4.7) in Australia, 2.4 (95% CI 1.3 to 4. 3) in Nigeria). CONCLUSION: Although there was a similar prevalence of atopy in both countries, Australian children had a higher prevalence of asthma symptoms. Further studies are needed to determine why atopic children in Australia are more at risk of developing asthma. Such studies will have important implications for the prevention of asthma. PMID- 10377206 TI - Remission of asthma in the middle aged and elderly: report from the Obstructive Lung Disease in Northern Sweden study. AB - BACKGROUND: Remission of asthma in adults has been considered to be low but is still not well documented. In children remission occurs with a rate estimated at approximately 50%. Remission of asthma in middle aged and elderly subjects was investigated as part of a population based study of respiratory diseases in Northern Sweden. METHODS: In 1986 86% of 6610 subjects participated in a questionnaire survey. After a clinical validation study 300 subjects were diagnosed as having current asthma. In 1996 5935 subjects of the cohort could be traced for a third survey and 87% participated. Of the subjects with current asthma in 1986, 267 participated. In addition, 60 symptomatic subjects were classified as suspected asthma and 58 of them participated in 1996. Remission of asthma was defined as no recurrent wheeze, no attacks of shortness of breath, and no use of asthma medicines in 1996. RESULTS: Remission of asthma during the 10 year period under study was 6%. In subjects with suspected asthma, remission occurred in 22%. The average annual remission rate was less than 1%. Remission was associated with previously mild disease and cessation of smoking. CONCLUSION: Remission of asthma or the disappearance of its symptoms to an asymptomatic latent phase appeared to be rare in middle aged and elderly subjects. PMID- 10377207 TI - Evaluation of the buccal component of systemic absorption with inhaled fluticasone propionate. AB - BACKGROUND: Inhaled corticosteroids have dose related systemic effects determined by oral (swallowed or oropharyngeal absorption) and lung bioavailability. A study was undertaken to evaluate the significance of oropharyngeal absorption for fluticasone propionate. METHODS: Sixteen healthy volunteers of mean age 29.3 years were studied using an open randomised, placebo controlled, four way crossover design. Treatments were: (a) fluticasone metered dose inhaler (pMDI) 250 microg, 8 puffs; (b) fluticasone pMDI 250 microg, 8 puffs + mouth rinsing/gargling (water); (c) fluticasone pMDI 250 microg, 8 puffs + mouth rinsing/gargling (charcoal); and (d) placebo pMDI, 8 puffs + mouth rinsing/gargling (water). Overnight (ONUC) and early morning (EMUC) urinary cortisol/creatinine ratios and 8 am serum cortisol (SC) levels were measured. RESULTS: Significant (p<0. 05) suppression of ONUC, EMUC, and SC occurred with each active treatment compared with placebo. The mean values (95% CI for difference from placebo) were: (a) ONUC (nmol/mmol): fluticasone (2. 8, 95% CI 3.6 to 7.9), fluticasone + water (3.1, 95% CI 3.3 to 7.7), fluticasone + charcoal (2.3, 95% CI 4.1 to 8.5); placebo (8.6); (b) EMUC (nmol/mmol): fluticasone (5.6, 95% CI 8.4 to 24.5), fluticasone + water (7.6, 95% CI 6.6 to 22.4); fluticasone + charcoal (5.6, 95% CI 8.7 to 24.5); placebo (22.1). There were no significant differences between active treatments. The numbers of subjects with an overnight urinary cortisol of <20 nmol/10 hours were 0 (placebo), 11 (fluticasone), 12 (fluticasone + water), and 13 (fluticasone + charcoal). CONCLUSIONS: Oropharyngeal absorption of fluticasone does not significantly contribute to its overall systemic bioactivity as assessed by sensitive measures of adrenal suppression. In view of almost complete hepatic first pass inactivation with fluticasone, there is no rationale to employ mouth rinsing to reduce its systemic effects although it may be of value for reducing oral candidiasis. PMID- 10377208 TI - Pulmonary lymphangioleiomyomatosis in Korea. AB - BACKGROUND: Pulmonary lymphangioleiomyomatosis (LAM) is a rare disease occurring in women of reproductive age and leading to progressive respiratory failure in spite of treatment. In Korea the first case was reported in 1984 and by 1997 a total of 23 cases had been reported. The clinical findings of these Korean cases are reviewed. METHODS: The details of 10 cases of LAM on file at Seoul National University Hospital were reviewed together with those of 13 cases previously reported from other Korean institutes. Two, including the only one to be reported in a man, were excluded after reviewing the clinical, radiological, and pathological findings, leaving a total of 21 cases in the present study. RESULTS: All 21 patients were women and in all cases the disease was proven pathologically. The mean (SD) age at onset of symptoms was 32 (8.6) years. The most common symptoms were dyspnoea and pneumothorax which were seen in 19 (90%) and 13 (76%) patients, respectively. Pulmonary function tests showed decreased transfer factor (TLCO) (100%) and airflow limitation (67%). All the cases had characteristic cysts on high resolution computed tomographic (HRCT) scanning. The overall severity score based on HRCT scans correlated with the percentage predicted TLCO/VA (p = 0.03) and FEV1/FVC (p = 0.02). The patients were all treated with medroxyprogesterone and/or tamoxifen. Follow up was possible in 10 cases. Two of these patients appeared to stabilise with no appreciable change clinically or in lung function on medroxyprogesterone and/or tamoxifen, but the remaining patients all deteriorated with two dying of respiratory insufficiency and one of infection following lung transplantation. CONCLUSIONS: As in other countries, in Korea LAM occurs exclusively in women and progresses despite hormonal treatment. PMID- 10377209 TI - Effect of a single dose of salmeterol on the increase in airway eosinophils induced by allergen challenge in asthmatic subjects. AB - BACKGROUND: The long acting beta2 agonist salmeterol is very effective in preventing asthmatic responses to specific stimuli, and this effect could theoretically be due to some anti-inflammatory property in addition to bronchodilator property. METHODS: The protective effect of a single dose of salmeterol (50 microg) on allergen induced early and late responses and on the associated airway inflammation was investigated in a double blind, placebo controlled, crossover study in 11 atopic asthmatic subjects. Eosinophil percentages and concentrations of eosinophil cationic protein (ECP) in peripheral blood and in hypertonic saline induced sputum were measured 24 hours after allergen inhalation. RESULTS: Salmeterol effectively inhibited both early and late asthmatic responses in comparison with placebo. Salmeterol also inhibited the increase in the percentage of eosinophils in the sputum 24 hours after allergen inhalation (median (range) baseline 6% (1-36), after placebo 31% (5-75), after salmeterol 12% (1-63)). However, the increase in both sputum and serum ECP concentrations 24 hours after allergen challenge was not affected by pretreatment with salmeterol. CONCLUSIONS: A single dose of salmeterol inhibits the allergen induced airway responses and the increase in sputum eosinophils after allergen challenge. PMID- 10377210 TI - Issues at the interface between primary and secondary care in the management of common respiratory disease.2: Are we too ready to diagnose asthma in children? PMID- 10377211 TI - Rare diseases.3: Wegener's granulomatosis. PMID- 10377213 TI - Idiopathic azygos vein aneurysm: a rare cause of mediastinal mass. AB - Venous aneurysm of the azygos arch is a very rare cause of mediastinal mass and is usually an incidental finding on chest radiography. Nowadays the diagnosis is made by non-invasive tests such as thoracic CT scanning and/or magnetic resonance imaging. The case is described of an asymptomatic woman in whom a mediastinal mass due to an azygos vein aneurysm was diagnosed by non-invasive procedures, the aetiology of which, in all probability, was idiopathic. PMID- 10377214 TI - Structure-antiviral activity relationship in the series of pyrimidine and purine N-[2-(2-phosphonomethoxy)ethyl] nucleotide analogues. 1. Derivatives substituted at the carbon atoms of the base. AB - A series of dialkyl esters of purine and pyrimidine N-[2-(phosphonomethoxy)ethyl] derivatives substituted at position 2, 6, or 8 of the purine base or position 2, 4, or 5 of the pyrimidine base were prepared by alkylation of the appropriate heterocyclic base with 2-chloroethoxymethylphosphonate diester in the presence of sodium hydride, cesium carbonate, or 1,8-diazabicyclo[5,4, 0]undec-7-ene (DBU) in dimethylformamide. Additional derivatives were obtained by the transformations of the bases in the suitably modified intermediates bearing reactive functions at the base moiety. The diesters were converted to the corresponding monoesters by sodium azide treatment, while the free acids were obtained from the diester by successive treatment with bromotrimethylsilane and hydrolysis. None of the PME derivatives in the pyrimidine series, their 6-aza or 3-deaza analogues, exhibited any activity against DNA viruses or retroviruses tested, except for the 5 bromocytosine derivative. Substitution of the adenine ring in PMEA at position 2 by Cl, F, or OH group decreased the activity against all DNA viruses tested. PMEDAP was highly active against HSV-1, HSV-2, and VZV in the concentration range (EC50) of 0.07-2 microg/mL. Also the 2-amino-6-chloropurine derivative was strongly active (EC50 = 0.1-0. 4 microg/mL) against herpes simplex viruses and (EC50 = 0.006-0.3 microg/mL) against CMV and VZV. PMEG was the most active compound of the whole series against DNA viruses (EC50 approximately 0.01-0.02 microg/mL), though it exhibited significant toxicity against the host cells. The base-modified compounds did not show any appreciable activity against DNA viruses except for 7-deazaPMEA (IC50 approximately 7.5 microg/mL) against HIV-1 and MSV. The neutral (diisopropyl, diisooctyl) diesters of PMEA were active against CMV and VZV, while the corresponding monoesters were inactive. The diisopropyl ester of the 2-chloroadenine analogue of PMEA showed substantially (10-100x) higher activity against CMV and VZV than the parent phosphonate. Also, the diisopropyl and diisooctyl ester of PMEDAP inhibited CMV and VZV, but esterification of the phosphonate residue did not improve the activity against either MSV or HIV. PMID- 10377215 TI - Fluorination of 3-(3-(piperidin-1-yl)propyl)indoles and 3-(3-(piperazin-1 yl)propyl)indoles gives selective human 5-HT1D receptor ligands with improved pharmacokinetic profiles. AB - It has previously been reported that a 3-(3-(piperazin-1-yl)propyl)indole series of 5-HT1D receptor ligands have pharmacokinetic advantages over the corresponding 3-(3-(piperidin-1-yl)propyl)indole series and that the reduced pKa of the piperazines compared to the piperidines may be one possible explanation for these differences. To investigate this proposal we have developed versatile synthetic strategies for the incorporation of fluorine into these ligands, producing novel series of 4-fluoropiperidines, 3-fluoro-4-aminopiperidines, and both piperazine and piperidine derivatives with one or two fluorines in the propyl linker. Ligands were identified which maintained high affinity and selectivity for the 5 HT1D receptor and showed agonist efficacy in vitro. The incorporation of fluorine was found to significantly reduce the pKa of the compounds, and this reduction of basicity was shown to have a dramatic, beneficial influence on oral absorption, although the effect on oral bioavailability could not always be accurately predicted. PMID- 10377212 TI - The molecular basis of asbestos induced lung injury. PMID- 10377216 TI - A peptide agonist acts by occupation of a monomeric G protein-coupled receptor: dual sites of covalent attachment to domains near TM1 and TM7 of the same molecule make biologically significant domain-swapped dimerization unlikely. AB - Membrane receptor dimerization is a well-established event for initiation of signaling at growth factor receptors and has been postulated to exist for G protein-coupled receptors, based on correction of nonfunctional truncated, mutant, or chimeric constructs by coexpression of appropriate normal complementary receptor domains. In this work, we have directly explored the molecular composition of the minimal functional unit of an agonist ligand and the wild-type G protein-coupled cholecystokinin (CCK) receptor, using photoaffinity labeling with a CCK analogue probe incorporating dual photolabile benzoylphenylalanine (Bpa) residues as sites of covalent attachment. This probe, 125I-D-Tyr-Gly-[(Nle28, 31, Bpa29,33)CCK-26-33], was shown to represent a full agonist and to specifically label the CCK receptor. Like probes incorporating individual photolabile residues in these positions,1,2 the two Bpa residues in the dual photoprobe covalently labeled receptor domains in the amino-terminal tail outside TM1 and in the third extracellular loop outside TM7. Absence of demonstrable receptor dimerization after the establishment of dual sites of covalent attachment supports the presence of these two domains within a single receptor molecule. Demonstration of the covalent adduct of a single probe molecule with the two cyanogen bromide fragments of the CCK receptor representing the expected domains further supports this interpretation. Thus, while domain swapped dimerization of G protein-coupled receptors may be possible as a mechanism of rescue for nonfunctional molecules, it is not necessary for ligand binding and initiation of signaling at a wild-type receptor in this superfamily. The functional unit for CCK action is normally a ligand-receptor monomer. PMID- 10377217 TI - Phthalein derivatives as a new tool for selectivity in thymidylate synthase inhibition. AB - A new set of phthalein derivatives stemming from the lead compound, phenolphthalein, were designed to specifically complement structural features of a bacterial form of thymidylate synthase (Lactobacillus casei, LcTS) versus the human TS (hTS) enzyme. The new compounds were screened for their activity and their specificity against TS enzymes from different species, namely, L. casei (LcTS), Pneumocystis carinii (PcTS), Cryptococcus neoformans (CnTS), and human thymidylate synthase (hTS). Apparent inhibition constants (Ki) for all the compounds against LcTS were determined, and inhibition factors (IF, ratio between the initial rates of the enzymatic reaction in the presence and absence of each inhibitor) against each of the four TS species were measured. A strong correlation was found between the two activity parameters, IF and Ki, and therefore the simpler IF was used as a screening factor in order to accelerate biological evaluation. Compounds 5b, 5c, 5ba, and 6bc showed substantial inhibition of LcTS while remaining largely inactive against hTS, illustrating for the first time remarkable species specificity among TSs. Due to sequence homology between the enzymes, several compounds also showed high activity and specificity for CnTS. In particular, 3-hydroxy-3-(3-chloro-4-hydroxyphenyl)-6-nitro-1H, 3H naphtho[1,8-c,d]pyran-1-one (6bc) showed an IF < 0.04 for CnTS (Ki = 0.45 microM) while remaining inactive in the hTS assay at the maximum solubility concentration of the compound (200 microM). In cell culture assays most of the compounds were found to be noncytotoxic to human cell lines but were cytotoxic against several species of Gram-positive bacteria. These results are consistent with the enzymatic assays. Intriguingly, several compounds also had selective activity against Cr. neoformans in cell culture assay. In general, the most active and selective compounds against the Gram-positive bacteria were those designed and found in the enzyme assay to be specific for LcTS versus hTS. The original lead compound was least selective against most of the cell lines tested. To our knowledge these compounds are the first TS inhibitors selective for bacterial TS with respect to hTS. PMID- 10377218 TI - Tricyclic farnesyl protein transferase inhibitors: crystallographic and calorimetric studies of structure-activity relationships. AB - Crystallographic and thermodynamic studies of farnesyl protein transferase (FPT) complexed with novel tricyclic inhibitors provide insights into the observed SAR for this unique class of nonpeptidic FPT inhibitors. The crystallographic structures reveal a binding pattern conserved across the mono-, di-, and trihalogen series. In the complexes, the tricycle spans the FPT active site cavity and interacts with both protein atoms and the isoprenoid portion of bound farnesyl diphosphate. An amide carbonyl, common to the tricyclic compounds described here, participates in a water-mediated hydrogen bond to the protein backbone. Ten high-resolution crystal structures of inhibitors complexed with FPT are reported. Included are crystallographic data for FPT complexed with SCH 66336, a compound currently undergoing clinical trials as an anticancer agent (SCH 66336, 4-[2-[4-(3,10-dibromo-8-chloro-6,11-dihydro-5H-benzo[5, 6]cyclohepta[1, 2-b]pyridin-11-yl)-1-piperidinyl]-2-oxoethyl]-1-piperidinecarbo xamide ). Thermodynamic binding parameters show favorable enthalpies of complex formation and small net entropic contributions as observed for 4-[2-[4-(3,10 dibromo-8-chloro-6,11-dihydro-11H-benzo[5, 6]cyclohepta[1, 2-b]pyridin-11 ylidene)-1-piperidinyl]-2-oxoethyl]pyridine N-oxide where DeltaH degrees bind = 12.5 kcal/mol and TDeltaS degrees bind = -1.5 kcal/mol. PMID- 10377219 TI - Design and synthesis of modified quinolones as antitumoral acridones. AB - The bacterial topoisomerase II (DNA gyrase) and the mammalian topoisomerase II represent the cellular targets for quinolone antibacterials and a wide variety of anticancer drugs, respectively. In view of the mechanistic similarities and sequence homologies exhibited by the two enzymes, tentative efforts to selectively shift from an antibacterial to an antitumoral activity was made by synthesizing a series of modified tricyclic quinolones, in which the essential 3 carboxylic function is surrogated by phenolic OH and the classic C-6 fluorine atom is replaced by a NH2 group. The resulting 7-amino-9-acridone derivatives were assayed for their antibacterial as well as cytotoxic activities. No antibacterial activity was found. On the other hand, many derivatives showed significant cytotoxic activity against both HL-60 and P388 leukemias and a wide panel of human and rodent solid tumor cells, derivatives 25 and 26 displaying the best overall antiproliferative activity. Against the LoVo cell line, derivative 25 exhibited higher cytotoxic effects than etoposide. PMID- 10377220 TI - Novel multidrug resistance reversal agents. AB - A series of 59 alpha-aryl-alpha-thioether-alkyl, -alkanenitrile, and alkanecarboxylic acid methyl ester tetrahydroisoquinoline and isoindoline derivatives (15a-48) were synthesized and evaluated as multidrug resistance (MDR) reversal agents. The compounds were tested on S1-B1-20 human colon carcinoma cells selected for resistance to bisantrene. Both the cytotoxicity of the reversal agents and their ability to resensitize the cells to bisantrene were determined. All but two of these compounds (15q, 40) were more effective MDR reversal agents in vitro than verapamil (VRP), a calcium channel antagonist which also has been shown to possess MDR modulating activity. Several showed good activity in this assay (IC50's < 0.5 microM), the most potent being isoindolines 44 (IC50 0.26 microM) and 46 (IC50 0.26 microM) and tetrahydroisoquinolines 47 (IC50 0.29 microM) and 15m (IC50 0.30 microM). A number of compounds were evaluated in vivo against vincristine (VCR)-resistant murine P388 leukemia, as well as against human epidermoid carcinoma KB/8.5 implanted sc in athymic mice. The reversal agents which consistently showed the highest activity, together with low toxicity, were alpha-aryl-alpha-thiotolylalkanenitrile tetrahydroisoquinoline derivatives with electron-rich alkoxy substituents on the aromatic rings. Of the tested compounds, the most effective reversal agents for both tumor lines were 15h (33% increased life span at 12.5 mg/kg, 0.2 mg/kg VCR versus VCR alone in the VCR-resistant P388 leukemia model and 59% relative tumor growth at 50 mg/kg, 8 mg/kg doxorubicin versus doxorubicin alone in the KB/8.5 model) and 39a (48% increased life span at 50 mg/kg, 0.2 mg/kg VCR versus VCR alone in the VCR resistant P388 leukemia model and 46% relative tumor growth at 25 mg/kg, 8 mg/kg doxorubicin versus doxorubicin alone in the KB/8.5 model). The mechanism of action of these compounds is believed to involve blocking the drug efflux pump, P glycoprotein. PMID- 10377221 TI - Butenolide endothelin antagonists with improved aqueous solubility. AB - Continued development around our ETA-selective endothelin (ET) antagonist 1 (CI 1020) has led to the synthesis of analogues with improved aqueous solubility profiles. Poor solubility characteristics displayed by 1 required a complex buffered formulation in order to conduct iv studies. To overcome the use of specific iv formulations for preclinical studies on additional drug candidates, analogues with improved aqueous solubility were desired. Several analogues were synthesized with substitution patterns that allowed for the formation of either acid or base addition salts. These derivatives had dramatically improved aqueous solubility. In addition, these analogues retained equivalent or improved ETA receptor selectivity and antagonist potency, versus 1, both in vitro and in vivo. Compound 29, which contains as a substituent the sodium salt of a sulfonic acid, has an ETA IC50 = 0.38 nM, ETA selectivity of 4200-fold, and ETA functional activity of KB = 7.8, all of which are similar or superior to those of 1. Compound 29 also has vastly superior aqueous solubility and solubility duration, compared to 1. Furthermore, 29 after iv infusion displays improved activity to 1 in preventing acute hypoxia-induced pulmonary hypertension in rats with an ED50 = 0.3 microg/kg/h. PMID- 10377222 TI - Prediction of ligand-receptor binding thermodynamics by free energy force field three-dimensional quantitative structure-activity relationship analysis: applications to a set of glucose analogue inhibitors of glycogen phosphorylase. AB - Glucose analogue inhibitors of glycogen phosphorylase, GP, may be of clinical interest in the regulation of glycogen metabolism in diabetes. The receptor geometry of glycogen phosphorylase b, GPb, is available for structure-based design and also for the evaluation of the thermodynamics of ligand-receptor binding. Free energy force field (FEFF) 3D-QSAR analysis was used to construct ligand-receptor binding models. FEFF terms involved in binding are represented by a modified first-generation AMBER force field combined with a hydration shell solvation model. The FEFF terms are then treated as independent variables in the development of 3D-QSAR models by correlating these energy terms with experimental binding energies for a training set of inhibitors. The genetic function approximation, employing both multiple linear regression and partial least squares regression data fitting, was used to develop the FEFF 3D-QSAR models for the binding process and to scale the free energy force field for this particular ligand-receptor system. The significant FEFF energy terms in the resulting 3D QSAR models include the intramolecular vacuum energy of the unbound ligand, the intermolecular ligand-receptor van der Waals interaction energy, and the van der Waals energy of the bound ligand. Other terms, such as the change in the stretching energy of the receptor on binding, change in the solvation energy of the system on binding, and the change in the solvation energy of the ligand on binding are also found in the set of significant FEFF 3D-QSAR models. Overall, the binding of this class of ligands to GPb is largely characterized by how well the ligand can sterically fit into the active site of the enzyme. The FEFF 3D QSAR models can be used to estimate the binding free energy of any new analogue in substituted glucose series prior to synthesis and testing. PMID- 10377223 TI - Design and synthesis of potent, selective, and orally bioavailable tetrasubstituted imidazole inhibitors of p38 mitogen-activated protein kinase. AB - Novel potent and selective diarylimidazole inhibitors of p38 MAP (mitogen activated protein) kinase are described which have activity in both cell-based assays of tumor necrosis factor-alpha (TNF-alpha) release and an animal model of rheumatoid arthritis. The SAR leading to the development of selectivity against c Raf and JNK2alpha1 kinases is presented, with key features being substitution of the 4-aryl ring with m-trifluoromethyl and substitution of the 5-heteroaryl ring with a 2-amino substituent. Cell-based activity was significantly enhanced by incorporation of a 4-piperidinyl moiety at the 2-position of the imidazole which also enhanced aqueous solubility. In general, oral bioavailability of this class of compounds was found to be poor unless the imidazole was methylated on nitrogen. This work led to identification of 48, a potent (p38 MAP kinase inhibition IC50 0.24 nM) and selective p38 MAP kinase inhibitor which inhibits lipopolysaccharide-stimulated release of TNF-alpha from human blood with an IC50 2.2 nM, shows good oral bioavailability in rat and rhesus monkey, and demonstrates significant improvement in measures of disease progression in a rat adjuvant-induced arthritis model. PMID- 10377224 TI - Synthesis of 9-O-substituted derivatives of 9-hydroxy-5, 6-dimethyl-6H-pyrido[4,3 b]carbazole-1-carboxylic acid (2-(dimethylamino)ethyl)amide and their 10- and 11 methyl analogues with improved antitumor activity. AB - Analogues of the antitumor drug S 16020-2 modified at the 9, 10, or 11 position were synthesized and evaluated in vitro and in vivo on the P388 leukemia and B16 melanoma models. Starting from 9-methoxy-5, 11-dimethyl-6H-pyrido[4,3-b]carbazole 1-carboxylic acid ethyl ester, the 11-CH3 analogue of 9-hydroxy-5,6-dimethyl-6H pyrido[4, 3-b]carbazole-1-carboxylic (2-(dimethylamino)ethyl)amide (1), compound 4, was synthesized using a four-step sequence, whereas its 10-CH3 analogue 5 was prepared using a two-step pathway, starting from compound 1. Finally starting from the 9-OH compounds 1, 4, and 5, a series of variously 9-O-substituted derivatives were synthesized. In these series, the most active compounds resulted from esterification of the 9-OH group with various aliphatic diacids, which led to 9-O-CO-( )-COOH derivatives of 1, 4, and 5. For these compounds, the number of long-term surviving mice obtained at the optimal dose were 60-100% in the ip/iv P388 leukemia and 10-35% in the ip/ip B16 melanoma, corresponding to an improved therapeutic index with respect to 1 and 4. This high antitumor activity, with curative examples in both models, was not due to a higher cytotoxicity since these compounds were equally or slightly less potent in vitro than 1 and 4. The most active compounds were thus selected for further in vivo evaluation. PMID- 10377225 TI - Stereoselective characterization of the 1,4-dihydropyridine binding site at L type calcium channels in the resting state and the opened/inactivated state. AB - Via a 3D-QSAR pseudoreceptor modeling approach, atomistic binding site models for pharmacologically active 1,4-dihydropyridines (DHPs) were developed. Applying a training set of pure DHP enantiomers a pseudoreceptor model representing the resting state of voltage-gated calcium channels (VGCCs) was generated by correlating experimental versus predicted free energies of binding (DeltaG degrees). For validation further test set derivatives-not used for receptor generation-were predicted yielding root-mean-square (rms) deviation of 0.532 kcal/mol. Selectivity of the resting state model was checked by using the same DHP training set compounds but experimental data for the inactivated channel mode. Although there was found an almost perfect correlation for the training set, the following free relaxation of the corresponding test set applying a Monte Carlo protocol showed rms of 2.033 kcal/mol, clearly demonstrating the lack of any predicting character of the hybrid model. Taking into consideration 19 additional nifedipine analogues, a further verification of the model was performed. This yielded a good correlation for the 12 training set compounds and a satisfactory prediction for the test set molecules with rms of 0.409 kcal/mol. The generation of a pseudoreceptor model depicting the opened/inactivated state of VGCCs required one single additional residue to achieve a rms of 0.848 kcal/mol for the prediction of the test set derivatives. Since all pseudoreceptor models are composed of the same six amino acid residues-Thr, Phe, Gly, Met, Tyr, Tyr-transition from resting to open/inactivated state may be described by one additional hydrogen bond donor interaction (Thr) at the left-hand side of DHPs. Furthermore, a potential charge-transfer interaction for all electron-deficient 4 phenyl DHPs is postulated, because significant correlation between quantum chemically AM1 (R = 0.91) and RHF 6-31G (R = 0.84) computed LUMO energies and experimentally detected DeltaG degrees exp values was found. PMID- 10377226 TI - Structure-activity relationships of L-dioxolane uracil nucleosides as anti Epstein Barr virus agents. AB - A series of 1,3-dioxolanyluracil analogues was prepared from the dioxolane intermediates 2, and their anti-Epstein Barr virus (anti-EBV) activities were determined. The potency of L-dioxolane uracil nucleosides against EBV replication is dependent on the substituents at the 5-position in the following decreasing order: I > Br > Cl > CH3 > CF3 > F. The most active and selective analogue was the iodo derivative (L-I-OddU) with an EC50 value of 0.03 microM and an EC90 value of 0.16 microM. There was no cytotoxicity or depletion of mitochondrial DNA in cells after exposure to L-I-OddU at 50 microM. The action against EBV replication in H1 cells is time-dependent, and EBV DNA in cells treated with L-I OddU could rebound to pretreatment levels once the drug was removed. In view of the potent antiviral activity plus favorable toxicity profiles, L-I-OddU may be potentially useful for the treatment of EBV-related infectious diseases as well as for delaying the onset or decreasing the incidence of EBV-associated cancers. PMID- 10377227 TI - Benzodiazepine receptor ligands. 4. Synthesis and pharmacological evaluation of 3 heteroaryl-8-chloropyrazolo[5,1-c][1,2,4] benzotriazine 5-oxides. AB - The synthesis of new 3-heteroaryl-8-chloropyrazolo[5,1-c][1,2, 4]benzotriazine 5 oxides and their binding activities at the central benzodiazepine receptor (BZR) are reported. The derivatives substituted at the 3-position with electron-rich five-membered rings, such as pyrrole 11, 2-thiophene 13c, or 3-thiophene 13d, showed good affinity values for BZR. In in vivo tests the 3-(thien-3-yl)-8 chloropyrazolo[5,1-c][1,2,4] benzotriazine 5-oxide (13d) showed selective anticonvulsant activity. PMID- 10377228 TI - Novel and potent 6-chloro-3-pyridinyl ligands for the alpha4beta2 neuronal nicotinic acetylcholine receptor. AB - 1-[(6-Chloro-3-pyridinyl)methyl]-2-imidazolidine (1), the N-desnitro metabolite of the major insecticide imidacloprid, is known to have similar potency to that of (-)-nicotine as an inhibitor of [3H](-)-nicotine binding at the rat recombinant alpha4beta2 neuronal nicotinic acetylcholine receptor (nAChR); IC50 values in the present study are 3.8 nM for (-)-nicotine, 6.0 nM for 1, and 155 nM for imidacloprid. Synthesis of new analogues of 1, modified only in the heterocyclic moiety (five-, six-, or seven-membered rings with NH, S, O, and CH2 substituents), gave compounds varying from 4-fold higher potency (2-iminothiazole analogue 10) to >6000-fold less active than (-)-nicotine. Other potent N-[(6 chloro-3-pyridinyl)methyl] compounds are those in which the heterocyclic imine is replaced with pyrrolidine (19) (IC50 9 nM) or trimethylammonium (22) (IC50 18 nM). A novel conversion of (-)-nicotine to its 6-chloro analogue increased the potency 2-fold. These 6-chloro-3-pyridinyl compounds are of interest as novel nAChR probes and potential metabolites of candidate insecticides. PMID- 10377230 TI - Cross-linking and sequence-specific alkylation of DNA by aziridinylquinones. 3. Effects of alkyl substituents. AB - The cytotoxicities and DNA cross-linking abilities of several alkyl-substituted diaziridinylquinones have been investigated. The cytotoxicities were determined in DT-diaphorase-rich (H460 and HT29) and -deficient (H596 and BE) cell lines. It was shown that the cytotoxicities in these cell lines correlated with the relative rates of reduction by the purified human enzyme and with the cross linking efficiencies. The rates of reduction by DT-diaphorase were more dependent on the structures of the compounds than the reduction potentials, as determined by cyclic voltammetry. A computer model was also used to explain high efficiency of cross-linking and the GNC sequence selectivity of the reduced methyl substituted diaziridinylquinones. PMID- 10377229 TI - New (sulfonyloxy)piperazinyldibenzazepines as potential atypical antipsychotics: chemistry and pharmacological evaluation. AB - A series of 2- or 8-trifluoromethylsulfonyloxy (TfO) and 2- or 8 methylsulfonyloxy (MsO) 11-piperazinyldibenzodiazepines, -oxazepines, and thiazepines were synthesized and evaluated in pharmacological models for their potential clozapine-like properties. In receptor binding assays, the 2-TfO analogues (18a, GMC2-83; 24, GMC3-06; and previously reported GMC1-169, 9a) of the dibenzazepines have profiles comparable to that of clozapine, acting on a variety of CNS receptors except they lack M1 receptor affinity. Introduction of 2 TfO to clozapine leads to compound 9e (GMC61-39) which has a similar binding profile as that of clozapine including having M1 receptor affinity. Interestingly, the MsO analogues, as well as the 8-TfO analogues, have no or weak dopaminergic and serotonergic affinities, but all 8-sulfonyloxy analogues do have M1 affinities. In behavioral studies performed to indicate the potential antipsychotic efficacy and the propensity to induce EPS, 2-TfO analogues blocked effectively the apomorphine-induced climbing in mice in a dose-dependent manner with ED50 values (mg/kg) of 2.1 sc for 9a, 1.3 po for 18a, 2.6 sc for 24, and 8.2 sc for 9e. On the other hand, they showed a clear dose separation with regard to their ED50 values (mg/kg) for indicating catalepsy in rats (>44 sc for 9a, 28 po for 18a, 30 sc for 24, and >50 sc for 9e, respectively), thus implicating a more favorable therapeutic ratio (K/A, ED50 climbing/ED50 catalepsy) in comparison with typical neuroleptics such as haloperidol and isoclozapine. Furthermore, compound 18a was also demonstrated to be an orally potent DA antagonist with an ED50 value of 0.7 mg/kg po in the ex vivo L-DOPA accumulation model. The present study contributes to the SAR of 11-piperazinyldibenzazepines, and the 2-TfO analogues of 11-piperazinyldibenzazepines are promising candidates as clozapine like atypical antipsychotics with low propensity to induce EPS. PMID- 10377231 TI - Synthesis and nicotinic acetylcholine receptor in vivo binding properties of 2 fluoro-3-[2(S)-2-azetidinylmethoxy]pyridine: a new positron emission tomography ligand for nicotinic receptors. AB - The lead compound of a new series of 3-pyridyl ethers, the azetidine derivative A 85380 (3-[(S)-2-azetidinylmethoxy]pyridine), is a potent and selective ligand for the human alpha4beta2 nicotinic acetylcholine receptor (nAChR) subtype. In vitro, the fluoro derivative of A-85380 (2-fluoro-3-[(S)-2-azetidinylmethoxy]pyridine or F-A-85380) competitively displaced [3H]cytisine or [3H]epibatidine with Ki values of 48 and 46 pM, respectively. F-A-85380 has been labeled with the positron emitter fluorine-18 (t1/2 (half-life) = 110 min) by no-carrier-added nucleophilic aromatic substitution by K[18F]F-K222 complex with (3-[2(S)-N-(tert butoxycarbonyl)-2-azetidinylmethoxy]pyridin-2-yl) tri methylammonium trifluoromethanesulfonate as a highly efficient labeling precursor, followed by TFA removal of the Boc protective group. The total synthesis time was 50-53 min from the end of cyclotron fluorine-18 production (EOB). Radiochemical yields, with respect to initial [18F]fluoride ion radioactivity, were 68-72% (decay corrected) and 49-52% (non-decay-corrected), and the specific radioactivities at EOB were 4-7 Ci/micromol (148-259 GBq/micromol). In vivo characterization of [18F]F-A-85380 showed promising properties for PET imaging of central nAChRs. This compound does not bind in vivo to alpha7 nicotinic or 5HT3 receptors. Moreover, its cerebral uptake can be modulated by the synaptic concentration of the endogenous ligand acetylcholine. The preliminary PET experiments in baboons with [18F]F-A-85380 show an accumulation of the radiotracer in the brain within 60 min. In the thalamus, a nAChR-rich area, uptake of radioactivity reached a maximum at 60 min (4% I.D./100 mL of tissue). [18F]F-A-85380 appears to be a suitable radioligand for brain imaging nAChRs with PET. PMID- 10377232 TI - 2,4-Diphenyl furan diamidines as novel anti-Pneumocystis carinii pneumonia agents. AB - Dicationic 2,4-bis(4-amidinophenyl)furans 5-10 and 2, 4-bis(4-amidinophenyl)-3,5 dimethylfurans 14 and 15 have been synthesized. Thermal melting studies revealed high binding affinity of the compounds to poly(dA-dT) and to the duplex oligomer d(CGCGAATTCGCG)2. All of the new compounds were effective against Pneumocystis carinii pneumonia in the immunosuppressed rat model with up to 200-fold increase in activity compared to the control compound pentamidine. No toxicity was noted for 5, 7-10 at the dose of 10 micromol/kg/d; however, the isopropyl analogue 7 showed toxicity comparable to pentamidine at the dosage of 20 micromol/kg/d. Dimethylation of the parent compound on the furan ring resulted in reduced activity and increased toxicity. PMID- 10377233 TI - Design and synthesis of novel quinoxaline-2,3-dione AMPA/GlyN receptor antagonists: amino acid derivatives. AB - PNQX (1,4,7,8,9,10-hexahydro-9-methyl-6-nitropyrido[3, 4-f]quinoxaline-2,3-dione) is a potent AMPA (IC50 = 0.063 microM) and GlyN (IC50 = 0.37 microM) receptor antagonist that was developed in our laboratories. While possessing a desirable in vitro and in vivo activity profile, this compound suffers from low aqueous solubility. In an effort to improve its potency and physical properties, we have designed and synthesized novel ring-opened analogues 4, 6, 9, and 11. Modeling analyses demonstrated that, while the 5-substituent in these analogues was forced to adopt an out-of-plane conformation due to steric contacts with neighboring substituents, the overall structure retained a good fit to a previously described AMPA pharmacophore model. This nonplanar orientation may lessen efficient packing in the solid state, compared to PNQX, leading to increased water solubility. Indeed, several nonplanar analogues containing appropriate functionalities, for example, the sarcosine analogue 9, were found to retain AMPA (IC50 = 0.14 microM) and GlyN (IC50 = 0.47 microM) receptor affinity and possess improved aqueous solubility compared to PNQX. The synthesis and the SAR of these compounds are discussed. PMID- 10377234 TI - Synthesis of classical and a nonclassical 2-amino-4-oxo-6-methyl-5-substituted pyrrolo[2,3-d]pyrimidine antifolate inhibitors of thymidylate synthase. AB - Compounds 2-5 were designed as potential antifolate nonpolyglutamatable inhibitors of thymidylate synthase (TS). These analogues are structurally related to 2-amino-4-oxo-5-substituted quinazolines and 2-amino-4-oxo-5-substituted pyrrolo[2, 3-d]pyrimidines which have shown excellent inhibition of TS and, for the quinazoline, significant promise as clinically useful antitumor agents. Compounds 2-4 were synthesized by appropriate amine exchange reactions on pivaloyl-protected 5-dimethylaminomethyl-substituted 6-methyl pyrrolo[2,3 d]pyrimidine 7 which in turn was obtained from the Mannich reaction of pivaloylated-6-methyl pyrrolo[2, 3-d]pyrimidine 6. In instances where the amine exchange reaction was sluggish, the Mannich base was quaternized with methyl iodide which afforded much faster exchange reaction with improved yields. For compound 5, 4-mercaptopyridine was used as the nucleophile and reacted with 7. The analogues 2-4 inhibited Lactobacillus casei (lc) TS and recombinant human (h) TS with IC50 in the 10(-4) to 10(-5) M range. Compound 5 inhibited lcTS and hTS 20% at 26 and 25 microM, respectively. In addition, compound 5 inhibited the growth of Pneumocystis carinii and Toxoplasma gondii cells in culture by 76% at 32 x 10(-6) M and 50% at 831 x 10(-6) M, respectively. PMID- 10377235 TI - Potent inhibition of steroid sulfatase activity by 3-O-sulfamate 17alpha benzyl(or 4'-tert-butylbenzyl)estra-1,3,5(10)-trienes: combination of two substituents at positions C3 and c17alpha of estradiol. AB - Steroid sulfates are precursors of hormones that stimulate androgen- and estrogen dependent cancers. Thus, steroid sulfatase, the enzyme that catalyzes conversion of DHEAS and E1S to the corresponding unconjugated steroids DHEA and E1, appears to be one of the key enzymes regulating the level of active androgenic and estrogenic steroids. Since 17alpha-substituted benzylestradiols and 3-O-sulfamate estrone (EMATE) represent two families of steroid sulfatase inhibitors that probably act through different mechanisms, we synthesized compounds 3-O-sulfamate 17alpha-benzylestradiol (4) and 3-O-sulfamate 17alpha-(tert-butylbenzyl)estradiol (5) that contain two kinds of substituents on the same molecule. In our enzymatic assay using a homogenate of human embryonal (293) cells transfected with steroid sulfatase, compounds 4 and 5 were found to be more potent inhibitors than already known steroid sulfatase inhibitors that have only a C17alpha-substituent or only a C3-sulfamate group (EMATE). The IC50 values of 4 and 5 were, respectively, 0.39 and 0.15 nM for the transformation of E1S to E1 and 4.1 and 1.4 nM for the transformation of DHEAS to DHEA. Compound 5 inhibited the steroid sulfatase activity in intact transfected (293) cell culture assays by inactivating the enzyme activity. Compound 5 also inactivates the steroid sulfatase activity at lower concentration than EMATE in microsomes of transfected (293) cells. In this assay, an excess of natural substrate E1S protects enzyme against inactivation by 5 or EMATE. Furthermore, the unsulfamoylated analogue of 5, compound 3, did not inactivate the steroid sulfatase. PMID- 10377236 TI - 2-Amino-4-benzyl-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridines: novel selective beta3-adrenoceptor agonists. AB - Trimetoquinol (TMQ, 7) is a potent nonselective beta-adrenoceptor (AR) agonist. Replacement of the catechol moiety of TMQ with a 2-aminothiazole group resulted in novel thiazolopyridine derivatives 9-11 which have been synthesized and evaluated for biological activity on human beta1-, beta2-, and beta3-AR. The Bischler-Napieralski reaction has been employed as a novel approach to construct the 2-amino-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridine ring system. Although in radioligand binding studies analogues 9 and 10 did not show selectivity toward beta3-AR, they exhibited a high degree of selective beta3-AR agonist activity in functional assays. Moreover, the beta3-AR agonist activity of the 2-aminothiazole derivatives is abolished by N-acetylation (analogue 11) or ring opening (analogue 25). This illustrates the importance of the intact 2-amino-4,5,6,7 tetrahydrothiazolo[5,4-c]pyridine ring for beta3-AR activity. PMID- 10377237 TI - The soluble sperm factor that causes Ca2+ release from sea-urchin (Lytechinus pictus) egg homogenates also triggers Ca2+ oscillations after injection into mouse eggs. AB - Cytosolic extracts of boar sperm contain a soluble phospholipase C (PLC) activity that induces Ca2+ release in sea-urchin (Lytechinus pictus) egg homogenates and an uncharacterized protein factor that causes Ca2+ oscillations when injected into mammalian eggs. In the present study we fractionated boar sperm extracts on three different FPLC chromatographic columns and found that the fractions that caused maximal Ca2+ release in sea-urchin egg homogenates were also the ones that triggered Ca2+ oscillations in mouse eggs. Our data suggests that the sperm factor which triggers Ca2+ oscillations in eggs contains a PLC and not the 33 kDa glucosamine deaminase previously suggested to be one its components. PMID- 10377238 TI - Acyl phosphatase activity of NO-inhibited glyceraldehyde-3-phosphate dehydrogenase (GAPDH): a potential mechanism for uncoupling glycolysis from ATP generation in NO-producing cells. AB - Treatment of glyceraldehyde-3-phosphate - dehydrogenase (GA (GAPDH) with the NO donors S-nitrosoglutathione, 3-morpholinosydnonimine or diethylamine NONOate (diethylamine diazeniumdiolate) in vitro, inhibited its dehydrogenase activity and induced its acyl phosphatase activity. NO-producing cells, in turn, exhibited reduced GAPDH activity, increased glycolysis, and decreased ATP content, synthesis and turnover. These cellular alterations could be explained by the uncoupling of glycolytic flux from substrate level phosphorylation by the acyl phosphatase activity of NO-modified GAPDH. PMID- 10377239 TI - A conserved RGD (Arg-Gly-Asp) motif in the transferrin receptor is required for binding to transferrin. AB - The transferrin receptor contains a highly conserved Arg-Gly-Asp (RGD) sequence in the C-terminal region where transferrin is thought to bind. RGD sequences are commonly involved in cell adhesion. This sequence is crucial for transferrin binding, suggesting possible evolutionary links between molecules mediating iron uptake and cell adhesion. PMID- 10377240 TI - A single point mutation leads to an instability of the hetero-octameric structure of yeast phosphofructokinase. AB - Yeast phosphofructokinase is an oligomeric enzyme whose detectable activity in vitro depends on its hetero-octameric structure. Here we provide data demonstrating that an alanine residue at positions 874 (for the PFK1-encoded alpha-subunit) or 868 (for the PFK2-encoded beta-subunit) is crucial to achieve this structure. Thus subunits carrying substitutions by either aspartate or lysine of this residue cause a lack of phosphofructokinase activity in vitro and signals of the subunits are poorly detectable in Western blots. Size-exclusion HPLC in conjunction with ELISA detection of the enzyme protein confirmed that no functional octamer is produced in such mutants. Our data suggest that the mutant subunits, not being assembled, tend to aggregate and subsequently become degraded. Substitution of the alanine by valine in either subunit leads to a reduction in specific activities, as expected from a conservative exchange. The kinetic data of the latter mutant revealed a higher affinity to the substrate fructose 6-phosphate, a lower extent of ATP inhibition and a lower degree of activation by fructose 2,6-bisphosphate. In addition, the affinity of mutants carrying a valine instead of an alanine in either the alpha- or the beta-subunit to fructose 2, 6-bisphosphate was increased. As no X-ray data on eukaryotic phosphofructokinases are available yet, our data provide the first evidence that a non-charge amino acid at position 874 or 868 is essential for the formation of the functional oligomer. This conclusion is substantiated by comparison with the structure of the well-known prokaryotic enzyme. PMID- 10377241 TI - Intramolecular chaperone and inhibitor activities of a propeptide from a bacterial zinc aminopeptidase. AB - An aminopeptidase from Aeromonas caviae T-64 was translated as a preproprotein consisting of three domains; a signal peptide (19 amino acid residues), an N terminal propeptide (101 residues) and a mature region (273 residues). We demonstrated that a proteinase, which was isolated from the culture filtrate of A. caviae T-64, activated the recombinant pro-aminopeptidase by removal of the majority of the propeptide. Using L-Leu-p-nitroanilide as a substrate, the processed aminopeptidase showed a large increase in kcat when compared with the unprocessed enzyme, whereas the Km value remained relatively unchanged. The similar Km values for the pro-aminopeptidase and the mature aminopeptidase indicated that the N-terminal propeptide of the pro-aminopeptidase did not influence the formation of the enzyme-substrate complex, suggesting the absence of marked conformational changes in the active domain. In contrast, the marked difference in kcat suggests a significant decrease in the energy of one or more of the transition states of the enzyme-substrate reaction coordinate. Moreover, we showed that the activity of the urea-denatured pro-aminopeptidase could be recovered by dialysis, whereas the activity of the urea-denatured mature aminopeptidase, which lacked the propeptide, could not. Further to this, the propeptide-deleted aminopeptidase formed an inclusion body in the cytoplasmic space in Escherichia coli and was not secreted at all. These results suggested that the propeptide of the pro-aminopeptidase acted as an intramolecular chaperone that was involved with the correct folding of the enzyme in vitro and was required for extracellular secretion in E. coli. PMID- 10377242 TI - Kinetic and inhibition studies on substrate channelling in the bifunctional enzyme catalysing C-terminal amidation. AB - A series of experiments has been conducted to investigate the possibility that substrate channelling might occur in the bifunctional forms of enzymes carrying out C-terminal amidation, a post-translational modification essential to the biological activity of many neuropeptides. C-terminal amidation entails sequential action by peptidylglycine mono-oxygenase (PAM, EC 1.14.17.3) and peptidylamidoglycolate lyase (PGL, EC 4.3.2.5), with the mono-oxygenase catalysing conversion of a glycine-extended pro-peptide into the corresponding alpha-hydroxyglycine derivative, which is then converted by the lyase into amidated peptide plus glyoxylate. Since the mono-oxygenase and lyase reactions exhibit tandem reaction stereospecificities, channelling of the alpha-hydroxy intermediate might occur, as is the case for some other multifunctional enzymes. Selective inhibition of the mono-oxygenase domain by competitive ester inhibitors, as well as mechanism-based mono-oxygenase inactivation by the novel olefinic inhibitor 5-acetamido-4-oxo-6-phenylhex-2-enoate (N-acetylphenylalanyl acrylate), has little to no effect on the kinetic parameters of the lyase domain of the AE from Xenopus laevis. Similarly, inhibition of the lyase domain by the potent dioxo inhibitor 2,4-dioxo-5-acetamido-6-phenylhexanoate has little effect on the activity of the monooxygenase domain in the bifunctional enzyme. A series of experiments on intermediate accumulation and conversion were also carried out, along with kinetic investigations of the reactivities of the monofunctional and bifunctional forms of PAM and PGL towards substrates and inhibitors. Taken together, the results demonstrate the kinetic independence of the mono-oxygenase and lyase domains, and provide no evidence for substrate channelling between these domains in the bifunctional amidating enzyme. PMID- 10377244 TI - Identification and expression of Pen c 2, a novel allergen from Penicillium citrinum. AB - The mould genus, Penicillium, is known to be a significant source of environmental aero-allergens. One important allergen from Penicillium citrinum, Pen c 2, has been identified by means of two-dimensional immunoblotting using IgE containing patients' sera. This novel allergen was cloned, sequenced and expressed in Escherichia coli. The cloned cDNA encodes a large 457-amino acid protein precursor containing a 16-amino acid signal peptide, a 120-amino acid propeptide and the 321-amino acid mature protein. Comparison of the Pen c 2 sequence with known protein sequences revealed shared high sequence similarities with two vacuolar serine proteases from Aspergillus niger and Saccharomyces cerevisiae. Asp-46, His-78 and Ser-244 were found to constitute the catalytic triad of the 39-kDa Pen c 2. The DNA coding for Pen c 2 was cloned into vector PQE-30 and expressed in E. coli as a His-tag fusion protein that bound serum IgE from Penicillium-allergic patients on immunoblots. Recombinant Pen c 2 could therefore be used effectively for diagnosis and also potentially for the treatment of mould-derived allergic disorders. PMID- 10377243 TI - Stimulation of Drosophila TrpL by capacitative Ca2+ entry. AB - Trp-like protein (TrpL, where Trp is transient receptor-potential protein) of Drosophila, a non-selective cation channel activated in photoreceptor cells by a phospholipase C-dependent mechanism, is thought to be a prototypical receptor activated channel. Our previous studies showed that TrpL channels are not activated by depletion of internal Ca2+ stores when expressed in Sf9 cells. Using fura-2 to measure cation influx via TrpL, and cell-attached patch recordings to monitor TrpL single-channel activity directly, we have found a thapsigargin induced increase in TrpL activity in the presence of extracellular bivalent cations, with Ca2+>Sr2+>> Ba2+. The increase in TrpL channel activity was blocked by concentrations of La3+ that completely inhibited endogenous capacitative Ca2+ entry (CCE), but have no effect on TrpL, suggesting that TrpL exhibits trans stimulation by cation entry via CCE. TrpL has two putative calmodulin (CaM) binding domains, designated CBS-1 and CBS-2. To determine which site may be required for stimulation of TrpL by the cytosolic free Ca2+ concentration ([Ca2+]i), a chimaeric construct was created in which the C-terminal domain of TrpL containing CBS-2 was attached to human TrpC1, a short homologue of Trp that is not activated by depletion of internal Ca2+ stores or by a rise in [Ca2+]i. This gain-of-function mutant, designated TrpC1-TrpL, exhibited trans-stimulation by Ca2+ entry via CCE. Examination of CaM binding in gel-overlay experiments showed that TrpL and the TrpC1-TrpL chimaera bound CaM, but TrpC1 or a truncated version of TrpL lacking CBS-2 did not. These results suggest that only CBS-2 binds CaM in native TrpL and that the C-terminal domain containing this site is important for trans-stimulation of TrpL by CCE. PMID- 10377245 TI - Identification of human complement Factor H as a ligand for L-selectin. AB - The selectin family of adhesion molecules (E-, P- and L-selectins) is involved in leukocyte recruitment to sites of inflammation and tissue damage. Recently it has been shown that L-selectin is involved not only in leukocyte tethering and rolling, but also plays an important role in leukocyte activation. For example, glycosylation-dependent cell-adhesion molecule 1 (GlyCAM-1), a known ligand for L selectin, has been shown to enhance beta2-integrin function. GlyCAM-1 is a secreted protein and is present in mouse serum at a concentration of approx. 1.5 microg/ml. There is no obvious GlyCAM-1 homologue in man and, to date, L-selectin ligand(s) from human serum have not been characterized. Therefore we have used L selectin affinity chromatography, followed by ion-exchange chromatography, to isolate specific ligand(s) for L-selectin. Using this procedure, we have isolated three major glycoproteins of apparent molecular masses 170 kDa, 70kDa and 50 kDa. The 170 kDa protein band was digested with trypsin and peptides were analysed by delayed extraction matrix-assisted laser desorption ionization MS and protein database searching. The 170 kDa protein was identified as the human complement protein Factor H. Human Factor H, isolated by a different method, was shown to bind specifically to L-selectin in the presence of CaCl2, and binding was inhibited by anti-L-selectin antibodies, fucoidan and lipopolysaccharide. Only a part of the purified Factor H preparation bound to immobilized L-selectin. The interaction of Factor H with leukocyte L-selectin was shown to induce the secretion of tumour necrosis factor-alpha (TNF-alpha). Pretreatment of Factor H with sialidase reduced both the binding of L-selectin to Factor H and the Factor H-induced L-selectin-mediated TNF-alpha secretion by leukocytes. Taken together, these results demonstrate that a post-translationally modified form of human plasma Factor H is a potential physiological ligand for L-selectin. PMID- 10377246 TI - Molecular cloning of the cDNA coding for mouse aldehyde oxidase: tissue distribution and regulation in vivo by testosterone. AB - The cDNA coding for mouse aldehyde oxidase (AO), a molybdoflavoprotein, has been isolated and characterized. The cDNA is 4347 nt long and consists of an open reading frame predicting a polypeptide of 1333 amino acid residues, with 5' and 3' untranslated regions of 13 and 335 nt respectively. The apparent molecular mass of the translation product in vitro derived from the corresponding cRNA is consistent with that of the monomeric subunit of the AO holoenzyme. The cDNA codes for a catalytically active form of AO, as demonstrated by transient transfection experiments conducted in the HC11 mouse mammary epithelial cell line. The deduced primary structure of the AO protein contains consensus sequences for two distinct 2Fe-2S redox centres and a molybdopterin-binding site. The amino acid sequence of the mouse AO has a high degree of similarity with the human and bovine counterparts, and a significant degree of relatedness to AO proteins of plant origin. Northern blot and in situ hybridization analyses demonstrate that hepatocytes, cardiocytes, lung endothelial or epithelial cells and oesophagus epithelial cells express high levels of AO mRNA. In the various tissues and organs considered, the level of AO mRNA expression is not strictly correlated with the amount of the corresponding protein, suggesting that the synthesis of the AO enzyme is under translational or post-translational control. In addition, we observed sex-related regulation of AO protein synthesis. In the liver of male animals, despite similar amounts of AO mRNA, the levels of the AO enzyme and corresponding polypeptide are significantly higher than those in female animals. Treatment of female mice with testosterone increases the amounts of AO mRNA and of the relative translation product to levels similar to those in male animals. PMID- 10377247 TI - A monomer-dimer equilibrium modulates the interaction of the sunflower homeodomain leucine-zipper protein Hahb-4 with DNA. AB - We have analysed the interaction of the sunflower homeodomain leucine-zipper (Hd Zip) protein Hahb-4 with DNA. The complete Hd-Zip domain from Hahb-4 was able to select specific sequences from a random oligonucleotide mixture that contained a 9-bp core with four fixed and five degenerate positions. Analysis of the binding of some of the selected sequences suggests that Hahb-4 preferentially binds the dyad-symmetrical sequence CAAT(A/T)ATTG. Single-nucleotide replacements at positions 1, 5 or 9 of this sequence produced a decrease in binding of 2-4-fold. DNA binding as a function of protein concentration was non-hyperbolic. This behaviour could be explained by an equation in which dimer formation is a pre requisite for DNA binding. A global dissociation constant (Kd) of 1.31x10(-14) M2 could be calculated. The removal of the leucine zipper promoted a change in specificity and a decrease in binding affinity (Kd=5. 03x10(-5) M). Mutation of Phe-20 of the homeodomain into Leu completely abolished DNA binding. The mutant protein, however, was able to inhibit DNA binding by the non-mutant form, presumably through the formation of heterodimers. The analysis of this inhibitory effect at different mutant concentrations allowed the estimation of the Kd for the dimer-monomer equilibrium [about (2-4)x10(-6) M]; from this, a Kd of 3-6x10( 9) M for the dimer-DNA complex could be estimated. The results obtained indicate that the formation of dimers is the main factor influencing the interaction of Hahb-4 with DNA. It is proposed that shifts in a dimer-monomer equilibrium could be used within the cell to modulate the interaction of this protein with target genes. PMID- 10377248 TI - Structural defects of a Pax8 mutant that give rise to congenital hypothyroidism. AB - Pax proteins are transcriptional regulators that play important roles during embryogenesis. These proteins recognize specific DNA sequences via a conserved element: the paired domain (Prd domain). The low level of organized secondary structure, in the free state, is a general feature of Prd domains; however, these proteins undergo a dramatic gain in alpha-helical content upon interaction with DNA ('induced fit'). Pax8 is expressed in the developing thyroid, kidney and several areas of the central nervous system. In humans, mutations of the Pax8 gene, which are mapped to the coding region of the Prd domain, give rise to congenital hypothyroidism. Here, we have investigated the molecular defects caused by a mutation in which leucine at position 62 is substituted for an arginine. Leu62 is conserved among Prd domains, and contributes towards the packing together of helices 1 and 3. The binding affinity of the Leu62Arg mutant for a specific DNA sequence (the C sequence of thyroglobulin promoter) is decreased 60-fold with respect to the wild-type Pax8 Prd domain. However, the affinities with which the wild-type and the mutant proteins bind to a non specific DNA sequence are very similar. CD spectra demonstrate that, in the absence of DNA, both wild-type Pax8 and the Leu62Arg mutant possess a low alpha helical content; however, in the Leu62Arg mutant, the gain in alpha-helical content upon interaction with DNA is greatly reduced with respect to the wild type protein. Thus the molecular defect of the Leu62Arg mutant causes a reduced capability for induced fit upon DNA interaction. PMID- 10377249 TI - Chemical cleavage of the overexpressed mitochondrial F1beta precursor with CNBr: a new strategy to construct an import-competent preprotein. AB - We have isolated a soluble import-competent 15 kDa N-terminal fragment of the overexpressed Nicotiana plumbaginifolia F1beta precursor of the ATP synthase (N15pF1beta). The isolation was achieved after chemical cleavage, with CNBr, of the insoluble precursor collected in inclusion bodies, followed by purification of the fragment using ion-exchange chromatography. The purity of the final product was estimated to be more than 99%. N15pF1beta contained a presequence of 54 amino acid residues (except for the N-terminal methionine residue) and 82 N terminal residues of the mature protein. N15pF1beta was shown to be imported into isolated potato tuber mitochondria and to be processed by the isolated mitochondrial processing peptidase (MPP) integrated into the cytochrome bc1 complex of the respiratory chain. Addition of N15pF1beta at micromolar concentrations resulted in the inhibition of import of F1beta precursor and alternative oxidase precursor, synthesized in vitro, into isolated mitochondria as well as the processing of these precursors catalysed by the isolated MPP-bc1 complex. N15pF1beta conjugated via a biotin link to avidin blocked import sites even after the reisolation of mitochondria and inhibited the import of the mitochondrial precursors, indicating that it can be used as a substrate for the generation of a stable translocation intermediate. Our results present a novel procedure for the production of an N-terminal fragment of the F1beta precursor that contains all information necessary for mitochondrial targeting and processing and that can be used for structural and functional studies of the mitochondrial protein import system. This procedure has a general value because it can be used for the production of chemical quantities of any mitochondrial import substrate and presequence peptide. PMID- 10377250 TI - Induction of the multispecific organic anion transporter (cMoat/mrp2) gene and biliary glutathione secretion by the herbicide 2,4,5-trichlorophenoxyacetic acid in the mouse liver. AB - The canalicular multispecific organic anion transporter, cMoat, is an ATP-binding cassette protein expressed in the canalicular domain of hepatocytes. In addition to the transport of endo- and xenobiotics, cMoat has also been proposed to transport GSH into bile, the major driving force of bile-acid-independent bile flow. We have shown previously that the herbicide 2,4,5-trichlorophenoxyacetic acid (2,4,5-T), a peroxisome-proliferator agent, significantly increases bile acid-independent bile flow in mice. On this basis, the effect of the herbicide on cMoat gene expression was studied. A 3.6-fold increase in cMoat mRNA levels and a 2.5-fold increase in cMoat protein content were observed in the liver of mice fed on a diet supplemented with 0.125% 2,4,5-T. These effects were due to an increased rate of gene transcription (3.9-fold) and were not associated with peroxisome proliferation. Significant increases in bile flow (2.23+/-0.39 versus 1.13+/-0.15 microl/min per g of liver; P<0.05) and biliary GSH output (7.40+/ 3.30 versus 2.65+/-0.34 nmol/min per g of liver; P<0.05) were observed in treated animals. The hepatocellular concentration of total glutathione also increased in hepatocytes of treated mice (10.95+/-0.84 versus 5.12+/-0.47 mM; P<0.05), because of the induction (2.4-fold) of the heavy subunit of the gamma-glutamylcysteine synthetase (GCS-HS) gene. This is the first model of co-induction of cMoat and GCS-HS genes in vivo in the mouse liver, associated with increased glutathione synthesis and biliary glutathione output. Our observations are consistent with the hypothesis that the cMoat transporter plays a crucial role in the secretion of biliary GSH. PMID- 10377251 TI - Molecular cloning of aryl-alcohol oxidase from the fungus Pleurotus eryngii, an enzyme involved in lignin degradation. AB - Aryl-alcohol oxidase (AAO), an extracellular enzyme characteristic of fungi from the genus Pleurotus, constitutes a source for H2O2 required in lignin biodegradation. The gene aao has been cloned, sequenced and characterized for the first time in Pleurotus eryngii. Both cDNA and genomic libraries were screened with probes obtained by PCR using as primers oligonucleotides corresponding to the N-terminus and internal sequences of AAO. DNA sequences from positive clones showed a unique open reading frame of 1779 nucleotides interrupted by 12 introns. The conceptual translation of the protein agrees with the partial amino acid sequences obtained from protein sequencing. A search for proteins with related amino-acid sequences revealed that glucose oxidase from Aspergillus niger has 33% identity and 51% similarity. A comparison with other oxidoreductases showed common motifs in both N- and C-terminal regions corresponding, respectively, to the FAD-binding region and the enzyme active site. However, AAO probably has structural differences with other oxidases, as deduced from its unique ability to generate H2O2 from the oxidation of aromatic alcohols. PMID- 10377252 TI - The N-terminal segment of endothelin-converting enzyme (ECE)-1b contains a di leucine motif that can redirect neprilysin to an intracellular compartment in Madin-Darby canine kidney (MDCK) cells. AB - Endothelin-converting enzyme (ECE)-1 is a membrane-bound metallopeptidase of the neprilysin (NEP) family. ECE-1 is responsible for the conversion of inactive big endothelins into active endothelins. Three different isoforms of human ECE-1 (ECE 1a, ECE-1b and ECE-1c) have been identified. They differ in their N-terminal cytosolic regions, have distinct tissue distribution and intracellular localization. ECE-1a and ECE-1c are both located at the cell surface whereas ECE 1b is targeted to an intracellular compartment. To better understand the nature of the signal responsible for the targeting of ECE-1b to the intracellular compartment, we have constructed several ECE/NEP chimaeric proteins and expressed them by transfection into Madin-Darby canine kidney (MDCK) cells. This allowed us to identify a nine amino acid segment in the cytosolic tail of ECE-1b that is sufficient to relocate NEP from the cell surface to an intracellular compartment. Site-directed mutagenesis on these chimaeras led to the identification of two leucine residues as part of the intracellular retention signal. PMID- 10377253 TI - Evidence that cyclophilin-A protects cells against oxidative stress. AB - Cyclophilin-A is the cytosolic isoform of a family of peptidylproline cis-trans isomerases that bind cyclosporin A. This study investigates the role of cyclophilin-A in necrotic cell death, induced by 'chemical ischaemia' and by t butylhydroperoxide. An 18-mer antisense phosphorothioate oligodeoxynucleotide was used to target a translated region of cyclophilin-A mRNA in rat neonatal cardiomyocytes. After a 24 h exposure to the oligonucleotide, the amount of cyclophilin-A in the cells was decreased by at least 93% as judged by immunological and enzymic criteria. For the enzyme assays, peptidyl proline cis trans-isomerase activity was measured fluorimetrically in small (10 microl) volumes of cell extract. Immunoblots were developed with a polyclonal anti cyclophilin-A antibody after sample isoelectric focusing and SDS/PAGE. Cyclophilin-A suppression had no effect on cyanide-plus-2-deoxyglucose-induced cell death. However, cyclophilin-A-suppressed cells were markedly more sensitive to t-butylhydroperoxide. Cyclosporin A conferred some resistance to the peroxide in both types of cell, but protection was greater in cyclophilin-A-suppressed cells, where cyclosporin A increased the survival time 2-fold. It is concluded that two cyclophilin isoforms are involved, in quite different ways, in peroxide induced cell death. Cyclophilin-A has a protective role. Another isoform, possibly mitochondrial cyclophilin-D, has a deleterious role, such that blockade by cyclosporin A leads to protection. PMID- 10377254 TI - Co-ordinate variations in methylmalonyl-CoA mutase and methionine synthase, and the cobalamin cofactors in human glioma cells during nitrous oxide exposure and the subsequent recovery phase. AB - We investigated the co-ordinate variations of the two cobalamin (Cbl)-dependent enzymes, methionine synthase (MS) and methylmalonyl-CoA mutase (MCM), and measured the levels of their respective cofactors, methylcobalamin (CH3Cbl) and adenosylcobalamin (AdoCbl) in cultured human glioma cells during nitrous oxide exposure and during a subsequent recovery period of culture in a nitrous oxide free atmosphere (air). In agreement with published data, MS as the primary target of nitrous oxide was inactivated rapidly (initial rate of 0.06 h(-1)), followed by reduction of CH3Cbl (to <20%). Both enzyme activity and cofactor levels recovered rapidly when the cells were subsequently cultured in air, but the recovery was completely blocked by the protein-synthesis inhibitor, cycloheximide. During MS inactivation, there was a reduction of cellular AdoCbl and holo-MCM activity (measured in the absence of exogenous AdoCbl) to about 50% of pre-treatment levels. When the cells were transferred to air, both AdoCbl and holo-MCM activity recovered, albeit more slowly than the MS system. Notably, the regain of the holo-MCM and AdoCbl was enhanced rather than inhibited by cycloheximide. These findings confirm irreversible damage of MS by nitrous oxide; hence, synthesis of the enzyme is required to restore its activity. In contrast, restoration of holo-MCM activity is only dependent on repletion of the AdoCbl cofactor. We also observed a synchronous fluctuation in AdoCbl and the much larger hydroxycobalamin pool during the inactivation and recovery phase, suggesting that the loss and repletion of AdoCbl reflect changes in intracellular Cbl homoeostasis. Our data demonstrate that the nitrous oxide-induced changes in MS and CH3Cbl are associated with reversible changes in both MCM holoactivity and the AdoCbl level, suggesting co-ordinate distribution of Cbl cofactors during depletion and repletion. PMID- 10377255 TI - An aromatic, but not a basic, residue is involved in the toxicity of group-II phospholipase A2 neurotoxins. AB - Ammodytoxins (Atxs) A, B and C are basic phospholipase A2s from Vipera ammodytes ammodytes snake venom, and they exhibit presynaptic toxicity. The most toxic is AtxA, followed by AtxC, its naturally occurring F124-->I/K128-->E mutant, which is 17 times less toxic. Two mutants of AtxA have been produced in bacteria and characterized. The specific enzymic activity of the K128-->E mutant on mixed phosphatidylcholine/Triton X-100 micelles is similar to that of the wild type. The K108-->N/K111-->N mutant, however, possesses 160% of the wild-type activity. Replacement of the two basic residues by uncharged, polar residues on the opposite side of the protein to the enzyme active site and interfacial adsorption surface results in increased enzymic activity at the water/lipid aggregate interface, due to a redistribution of electrostatic charge. The binding affinity of the double mutant for the specific acceptor in bovine brain was similar to that of AtxA, whereas the affinity of the single mutant was similar to that of AtxC, which was slightly weaker than that of AtxA. Interestingly, the substitution of any of these three basic surface residues did not significantly change the lethal potency of AtxA. Since the single mutant AtxA(K128-->E) is equivalent to the AtxC(I124-->F) mutant, this indicates that the residue at position 124 is important for presynaptic toxicity of Atxs. The more than 10-fold lower toxicity of AtxC, compared with AtxA, is a consequence of the substitution of Phe-124 (aromatic ring) with Ile (aliphatic chain). Exposed aromatic residues in the C-terminal region may also be important for the neurotoxicity of other similar toxins. PMID- 10377256 TI - The DmpA aminopeptidase from Ochrobactrum anthropi LMG7991 is the prototype of a new terminal nucleophile hydrolase family. AB - The DmpA (d-aminopeptidase A) protein produced by Ochrobactrum anthropi hydrolyses p-nitroanilide derivatives of glycine and d-alanine more efficiently than that of l-alanine. When regular peptides are utilized as substrates, the enzyme behaves as an aminopeptidase with a preference for N-terminal residues in an l configuration, thus exemplifying an interesting case of stereospecificity reversal. The best-hydrolysed substrate is l-Ala-Gly-Gly, but tetra- and penta peptides are also efficiently hydrolysed. The gene encodes a 375-residue precursor, but the active enzyme contains two polypeptides corresponding to residues 2-249 (alpha-subunit) and 250-375 (beta-subunit) of the precursor. Residues 249 and 250 are a Gly and a Ser respectively, and various substitutions performed by site-directed mutagenesis result in the production of an uncleaved and inactive protein. The N-terminal Ser residue of the beta-subunit is followed by a hydrophobic peptide, which is predicted to form a beta-strand structure. All these properties strongly suggest that DmpA is an N-terminal amidohydrolase. An exploration of the databases highlights the presence of a number of open reading frames encoding related proteins in various bacterial genomes. Thus DmpA is very probably the prototype of an original family of N-terminal hydrolases. PMID- 10377257 TI - Chloroplast thioredoxin mutants without active-site cysteines facilitate the reduction of the regulatory disulphide bridge on the gamma-subunit of chloroplast ATP synthase. AB - The activity of the chloroplast H+-ATPase (CFoCF1) is regulated by the proton electrochemical membrane potential and the reduction or the formation of the disulphide bridge on the gamma-subunit mediated by chloroplast thioredoxins (Trx). The latter regulation also applies to the water-soluble portion of CFoCF1 (CF1) and includes two successive steps, namely the binding of Trx to CF1 and the subsequent reduction or oxidation of CF1. To study this process thoroughly, a new expression system for spinach Trx-f and Trx-m was designed. In the presence of dithiothreitol (DTT) both forms of the expressed Trx could reduce the disulphide bridge on the gamma-subunit of CF1 and thus activate the ATPase. Trx mutants deficient in the internal, or both, cysteines of the active site were designed to study the details of the interaction. The Trx mutant proteins could still activate CF1-ATPase in the presence of DTT and they also increased the apparent affinity of CF1 for DTT. This implies that the binding of Trx to the CF1 gamma subunit induces a conformational change facilitating the reduction of the disulphide bridge, and partially explains the high efficiency of Trx as a reductant in vivo. PMID- 10377258 TI - Purification, characterization and cDNA cloning of a phospholipase A2 inhibitor from the serum of the non-venomous snake Elaphe quadrivirgata. AB - The serum of a non-venomous striated snake, Elaphe quadrivirgata, was found to contain phospholipase A2 (PLA2) inhibitory proteins (PLIs). One of these inhibitors was purified by Sephadex G-200 gel filtration, Q-Sepharose FF ion exchange chromatography and Butyl Sepharose 4FF hydrophobic chromatography. The purified PLI inhibited the enzymic activities of all PLA2 groups, including Elapidae venom (group-I), Viperidae venom (group-II) and honeybee PLA2s (group III). The inhibitor was a 130 kDa glycoprotein consisting of two distinct subunits, A and B, of 30 and 29 kDa respectively; each of which was glycosylated with N-linked oligosaccharide chains. The cDNAs encoding the respective inhibitor subunits were isolated from a liver cDNA library by the use of probes, prepared by PCR, based on the partially determined amino-acid sequences of the corresponding subunits. The respective nucleotide sequences encoded 19-amino-acid residue signal sequences, followed by 183- and 181-residue protein sequences for the A and B subunits respectively. The amino-acid sequences revealed that the E. quadrivirgata inhibitor corresponded to PLIgamma, one of three kinds of inhibitors purified from venomous snakes. The existence of PLIgamma in the serum of this non-venomous snake suggested that, besides having a protective role against the venom PLA2s of other venomous snakes, PLIgamma has other important physiological functions in regulating local PLA2 activities; and thus it raises the possibility that PLIgamma occurs in other animals, including mammals. PMID- 10377259 TI - The influence of epitope availability on atomic-force microscope studies of antigen-antibody interactions. AB - The ability of the atomic-force microscope (AFM) to detect interaction forces between individual biological molecules has recently been demonstrated. In this study, force measurements have been obtained between AFM probes functionalized with the beta-subunit of human chorionic gonadotrophin (betahCG) and surfaces functionalized with anti-betahCG antibody. A comparison of the obtained results with previous anti-ferritin antibody-binding data identifies differences when the antigen molecule expresses only a single epitope (betahCG), rather than multiple epitopes (ferritin), for the monoclonal antibodies employed. Specifically, the probability of observing probe-sample adhesion is found to be higher when the antigen expresses multiple epitopes. However, the periodic force observed in the adhesive-force distribution, due to the rupture of single antigen-antibody interactions, is found to be larger and more clearly observed for the mono epitopic system. Hence, these findings indicate the potential of the AFM to distinguish between multivalent and monovalent antibody-antigen interactions, and demonstrate the influence of the number of expressed epitopes upon such binding studies. PMID- 10377260 TI - Doc2 is not associated with known regulated exocytotic or endosomal compartments in adrenal chromaffin cells. AB - Doc2 is a C2-domain-containing protein that is highly expressed in the nervous system and has a constitutively expressed isoform. It has been implicated as a potential Ca2+ sensor in regulated exocytosis, and has been suggested to be associated with synaptic vesicles. To examine whether Doc2 is associated with synaptic-like microvesicles (SLMVs) or dense-core granules in neuroendocrine cells, we examined the distribution of Doc2 in subcellular fractionation of chromaffin cells of the adrenal medulla and in PC12 cells. Doc2 did not co distribute with SLMVs from either cell type, but did appear to co-distribute with dense-core granules from PC12 cells. In contrast, it was not associated with the dense-core granules during subcellular fractionation of the adrenal medulla, and nor did it appear to be associated with endosomes, cis-Golgi or the trans-Golgi network. In contrast, Doc2 co-distributed under all conditions with a mitochondrial marker. We conclude that Doc2 is not a general component of regulated secretory vesicles, but may instead be associated with mitochondria. PMID- 10377261 TI - ADP ribosylation factor 1 mutants identify a phospholipase D effector region and reveal that phospholipase D participates in lysosomal secretion but is not sufficient for recruitment of coatomer I. AB - The small GTP-binding protein, ADP-ribosylation factor 1 (ARF1) is essential for the formation of coatomer-coated vesicles from the Golgi and is also an activator of phospholipase D (PLD). Moreover, ARF1-regulated PLD is part of the signal transduction pathway that can lead to secretion. In this study, substitution and deletion mutants of ARF1 were tested for their ability to activate PLD. These map the PLD effector region of ARF1 to the alpha2 helix, part of the beta2-strand and the N-terminal helix and its ensuing loop. ARF mutants with an increased or decreased ability to activate PLD showed similar characteristics when tested for their ability to stimulate secretion from HL60 cells. ARF1, deleted of the N terminal 17 amino acid residues (Ndel17), did not support PLD activity or secretion, and neither did it inhibit the activity of wild-type myristoylated ARF1 (myrARF1). In contrast, Ndel17 effectively competed with wild-type myrARF1 to prevent coatomer binding to membranes. This appears to define a structural role for Ndel17, as it can bind a high-molecular mass complex in cytosol. In addition, ethanol has no effect on recruitment of coatomer to membrane. We conclude that the function of ARF-regulated PLD is in the signal-transduction pathway leading to secretion of lysosomal granules, and not as an essential component of ARF1-mediated coatomer binding. PMID- 10377262 TI - Processing of normal lysosomal and mutant N-acetylgalactosamine 4-sulphatase: BiP (immunoglobulin heavy-chain binding protein) may interact with critical protein contact sites. AB - The lysosomal hydrolase N-acetylgalactosamine-4-sulphatase (4-sulphatase) is essential for the sequential degradation of the glycosaminoglycans, dermatan and chondroitin sulphate and, when deficient, causes the lysosomal storage disorder mucopolysaccharidosis type VI. The cysteine at codon 91 of human 4-sulphatase was identified previously as a key residue in the active site of the enzyme and was mutated by site-directed mutagenesis to produce a 4-sulphatase in which cysteine 91 was replaced by a threonine residue (C91T). The C91T mutation caused a loss of 4-sulphatase activity, a detectable protein conformational change and a lower level of intracellular 4-sulphatase protein [Brooks, Robertson, Bindloss, Litjens, Anson, Peters, Morris and Hopwood (1995) Biochem. J. 307, 457-463]. In the present study, we report that C91T is synthesized normally in the endoplasmic reticulum as a 66 kDa glycosylated protein, which is very similar in size to wild type 4-sulphatase. However, C91T neither underwent normal Golgi processing, shown by lack of modification to form mannose 6-phosphate residues on its oligosaccharide side chains, nor did it traffic to the lysosome to undergo normal endosomal-lysosomal proteolytic processing. Instead, C91T remained in an early biosynthetic compartment and was degraded. The molecular chaperone, immunoglobulin binding protein (BiP), was associated with newly-synthesized wild type and mutant 4-sulphatase proteins for extended periods, but no direct evidence was found for involvement of BiP in the retention or degradation of the C91T protein. This suggested that prolonged association of mutant protein with BiP does not necessarily infer involvement of BiP in the quality control process, as previously implied in the literature. The predicted BiP binding sites on 4 sulphatase map to beta-strands and alpha-helices, which are co-ordinated together in the folded molecule, indicating that BiP interacts with critical protein folding or contact sites on 4-sulphatase. PMID- 10377263 TI - Characterization of the Ca2+-dependent binding of annexin IV to surfactant protein A. AB - We have shown previously that surfactant protein A (SP-A) binds to annexin IV in a Ca2+-dependent manner [Sohma, Matsushima, Watanabe, Hattori, Kuroki and Akino (1995) Biochem. J. 312, 175-181]. Annexin IV is a member of the annexin family having four consensus repeats of about 70 amino acids and a unique N-terminal tail. In the present study, the functional site of both annexin IV and SP-A for the Ca2+-dependent binding was investigated using mutant proteins. SP-A bound in a Ca2+-dependent manner to an annexin-IV truncation mutant consisting of the N terminal domain and the first three domains (T(N-1-2-3)). SP-A also bound to T3 4, but this interaction was not Ca2+-dependent. SP-A bound weakly to the other truncation mutants (T(N-1-2), T(2-3) and T(2-3-4)). Each consensus repeat of annexin IV possesses a conserved acidic amino acid residue (Glu70, Asp142, Glu226 and Asp301) that putatively ligates Ca2+. Using annexin-IV DE mutants in which one, two or three residues out of the four Asp/Glu were altered to Ala by site directed mutagenesis [Nelson and Creutz (1995) Biochemistry 34, 3121-3132], it was revealed that Ca2+ binding in the third domain is more important than in the other Ca2+-binding sites. SP-A is a member of the animal lectin group homologous with mannose-binding protein A. The substitution of Arg197 of rat SP-A with Asp or Asn eliminated binding to annexin IV, whereas the substitution of Glu195 with Gln was silent. These results suggest that the Ca2+ binding to domain 3 of annexin IV is required for the Ca2+-dependent binding by SP-A and that Arg197 of SP-A is important in this binding. PMID- 10377264 TI - Identification of Tyr-703 and Tyr-936 as the primary association sites for Grb2 and Grb7 in the c-Kit/stem cell factor receptor. AB - In this paper we demonstrate the presence of two novel in vivo autophosphorylation sites in the c-Kit/stem cell factor receptor (c-Kit/SCFR): Tyr-703 in the kinase insert and Tyr-936 in the C-terminal tail. We furthermore demonstrate that the adapter protein Grb2 is a specific binding partner for both phosphorylated Tyr-703 and phosphorylated Tyr-936, whereas the adapter protein Grb7 binds selectively to phosphorylated Tyr-936. It is shown that the association occurs through the Src homology 2 (SH2) domains of Grb2 and Grb7. Binding of Grb2 to Tyr-703 in the c-Kit/SCFR provides a link to the Ras/mitogen activated protein kinase pathway. PMID- 10377267 TI - Oscillatory dynamics and information processing in olfactory systems AB - Oscillatory dynamics is a universal design feature of olfactory information processing systems. Recent results in honeybees and terrestrial slugs suggest that oscillations underlie temporal patterns of olfactory interneuron responses critical for odor discrimination. Additional general design features in olfactory information-processing systems include (1) the use of central processing areas receiving direct olfactory input for odor memory storage and (2) modulation of circuit dynamics and olfactory memory function by nitric oxide. Recent results in the procerebral lobe of the terrestrial slug Limax maximus, an olfactory analyzer with oscillatory dynamics and propagating activity waves, suggest that Lucifer Yellow can be used to reveal a band-shaped group of procerebral neurons involved in the storage of an odor memory. A model has been constructed to relate wave propagation and odor memory bands in the procerebral lobe of L. maximus and to relate these findings to glomerular odor representations in arthropods and vertebrates. PMID- 10377265 TI - Angiotensin II stimulates serine phosphorylation of the adaptor protein Nck: physical association with the serine/threonine kinases Pak1 and casein kinase I. AB - Nck is a small adaptor protein consisting exclusively of three SH3 domains and one SH2 domain. Nck is thought to have an important role in cell signalling by coupling receptor tyrosine kinases, via its SH2 domain, to downstream SH3-binding effectors. We report here that angiotensin II, working through the AT1 receptor subtype, stimulates the phosphorylation of Nck in rat aortic smooth muscle cells. Phosphopeptide mapping analysis revealed that Nck is phosphorylated on four peptides containing exclusively phosphoserine in quiescent cells. Treatment with angiotensin II resulted in increased phosphorylation of these four peptides, without the appearance of new phosphopeptides. We show that Nck, via its SH3 domains, specifically binds three major phosphoproteins of 95, 82 and 66 kDa both in vitro and in intact cells. Notably, the phosphorylation of these Nck-binding proteins was found to increase in parallel with that of Nck on stimulation by angiotensin II. One candidate for the 66 kDa phosphoprotein is the serine/threonine kinase p21-activated kinase 1 (Pak1), which was found to form a stable complex with Nck in aortic smooth muscle cells. We have also identified the gamma2 isoform of casein kinase I as another protein kinase that associates with Nck in these cells. These findings indicate that Nck is a target of G protein-coupled receptors and suggest a role for Pak1 and casein kinase I-gamma2 in downstream signalling or regulation of the AT1 receptor. PMID- 10377268 TI - Tympanal hearing in the sarcophagid parasitoid fly Emblemasoma sp.: the biomechanics of directional hearing. AB - In Diptera, tympanal hearing has evolved at least twice in flies that belong to two different families, the tachinids and the sarcophagids. Common to these flies is their parasitoid reproductive strategy, both relying on the acoustic detection and localization of their hosts, singing insects, by means of tympanal hearing organs. In the present study, the external anatomy of the unusual hearing organs of the sarcophagid fly Emblemasoma sp. is described. The sarcophagid ears bear numerous anatomical similarities with those of ormiine tachinids: they are located on the ventral prosternum and possess a pair of scolopidial mechanoreceptive sense organs. A striking difference, however, resides in the lack of a well-defined presternum in the sarcophagid tympanal system. Instead, a deep longitudinal fold, the tympanal fold, spans both hemilateral tympanal membranes across the midline of the animal. Measured using laser Doppler vibrometry, the tympanal mechanical response in the sound field reveals asymmetrical deflection shapes that differ from those of tachinids. Lacking a central fulcrum, the sarcophagid tympanal complex presents different vibrational modes that also result in interaural coupling. The evolutionarily convergent, yet distinct, solutions used by these two small auditory systems to extract directional cues from the sound field and the role of tympanal coupling in this process are discussed. PMID- 10377269 TI - Ixodid ticks avoid contact with liquid water. AB - Larvae of the cattle tick Boophilus microplus and all life stages of the European sheep tick Ixodes ricinus avoid walking on a wet membrane surface surrounding a dry patch. Of 170 reactions made at a border with liquid water by 22 B. microplus larvae, 40% consisted of immediate turns to the opposite side to bring all the legs back onto a dry patch, 41% were walks along the border, during which the ticks maintained contact with both the dry and wet zones, and 19% were returns to the dry patch after a short excursion onto the wet surround. Since contact with one front leg tip was sufficient to cause return reactions from the wet surface in most of the border contacts, the water receptor(s) that enable ticks to perceive the wet surface are probably located in terminal pore sensilla on the first-leg tarsi. Observations on the return reactions of ticks with different groups of chemosensilla masked confirmed this. Ticks have an ambiguous relationship with water: they appear to avoid direct contact with it, but they need a high humidity to compensate for any deficit in body water. PMID- 10377266 TI - Induction of gadd153 mRNA by nutrient deprivation is overcome by glutamine. AB - The growth arrest and DNA damage-inducible (gadd) genes are co-ordinately activated by a variety of genotoxic agents and/or growth-cessation signals. The regulation of gadd153 mRNA was investigated in renal proximal tubular epithelial cells (LLC-PK1) cultured in a nutrient- and serum-deprived medium. The addition of glutamine alone to LLC-PK1 cells cultured in Earl's balanced salt solution (EBSS) is sufficient to suppress gadd153 mRNA expression, and the removal of only glutamine from Dulbecco's modified Eagle's medium (DMEM) is also sufficient to induce gadd153 mRNA expression. Consistent with these findings, the inhibition of glutamine utilization with acivicin and 6-diazo-5-oxo-l-norleucine (DON) in cells grown in a glutamine-containing medium effectively induces gadd153 expression. Glutamine can be used as an energy source in cultured mammalian cells. However, it is unlikely that deficits in cellular energy stores (ATP) are coupled to gadd153 mRNA expression, because concentrations of ATP, UTP and GTP are all elevated in EBSS-exposed cells, and the addition of alpha-oxoglutarate to cells grown in EBSS has no effect on gadd153 mRNA expression. In contrast, concentrations of CTP decline substantially in EBSS and glutamine-deprived DMEM cultured cells. Glutamine also serves as a precursor for the synthesis of protein and DNA. The addition of glutamine to cells grown in EBSS partly restores CTP concentrations. The addition of pyrimidine ribonucleosides (cytidine and uridine) to LLC-PK1 cells also restores CTP concentrations, in a manner commensurate with their relative abilities to overcome gadd153 expression. Finally, glutamine does not completely suppress DNA damage-induced gadd153 expression, suggesting that multiple signalling pathways lead to the expression of gadd153 mRNA under conditions of nutrient deprivation and DNA damage. PMID- 10377270 TI - Developmental analysis of Ganaspis xanthopoda, a larval parasitoid of Drosophila melanogaster. AB - Ganaspis xanthopoda is a solitary larval parasitoid wasp of the fruit fly Drosophila melanogaster. The life cycle of Ganaspis xanthopoda in the wild-type and developmental mutant ecdysoneless strains of Drosophila melanogaster is described. The female infects a second-instar host larva. The parasitoid embryo hatches into a mobile first-instar (L1) larva. The L1 parasitoid has fleshy appendages and, while mobile, it remains confined within the wandering larval host. The second-instar larva (L2) is an endoparasite within the host prepupa and lacks appendages. The L2-to-L3 molt is dependent on pupation and marks the transition of the endoparasite into an ectoparasite. The third-instar larva (L3) is a sessile ectoparasite, develops an extensive tracheal system and consumes the host as it progresses through its prepupal and pupal stages. A single adult male or female emerges from the host puparium. The developmental analysis of Ganaspis xanthopoda reveals a tight synchrony between host and parasitoid development which is, at least in part, dependent on the ecdysone levels of the host. PMID- 10377271 TI - Spatio-temporal learning by the ant ectatomma ruidum AB - We tested, under field and laboratory conditions, whether the neotropical ant Ectatomma ruidum Roger can learn several associations between temporal and spatial changes in the daily pattern of food availability. Honey was shuffled between two or three feeding sites following a fixed daily schedule. Foragers learnt to associate particular sites with the specific times at which food was available, individually marked ants being observed on the correct sites at the correct times. Some ants anticipated the time of food delivery by approximately 30 min, and it was not necessary for them to be rewarded at the first stage of the sequence of food collection to continue their search for honey according to the correct schedule of reward. Ants also followed the same schedule when no honey was supplied at each stage of the sequence, and they stayed at the expected unrewarded site for a period equivalent to the reward period of the corresponding training phase, indicating that they had learnt when and for how long the food was available. Thus, ants rely on their spatio-temporal memory rather than on local cues coming from the honey source to guide them. PMID- 10377272 TI - Functional complementation of the malvolio mutation in the taste pathway of Drosophila melanogaster by the human natural resistance-associated macrophage protein 1 (Nramp-1). AB - The malvolio (mvl) gene of Drosophila melanogaster encodes a protein with a high degree of homology to natural resistance-associated macrophage proteins (Nramps). This family of integral membrane proteins, many of which appear to function as cation transporters, is remarkably conserved in several phylogenetically distinct species. In Drosophila melanogaster, the protein Mvl is expressed in macrophages and in differentiated neurons; loss-of-function mutations lead to defects in gustatory behaviour. The human Nramp-1 protein was expressed in Drosophila melanogaster using the hsp70 promoter. Overexpression in normal animals does not lead to any alterations in their behaviour or physiology. In mutants, however, ubiquitous expression of human Nramp-1 can totally rescue the taste defect. This finding that Nramp-1 can complement the taste defect in mvl mutants provides a potent means of exploiting behavioural genetics to dissect the function of Nramp 1 and to identify other molecules involved with this transport system. PMID- 10377274 TI - The photoreceptors and visual pigments in the retina of a boid snake, the ball python (Python regius) AB - The photoreceptors and visual pigments of Python regius were studied using microspectrophotometry and scanning electron microscopy. The retina contains rods and cones, with rods constituting at least 90 % of the photoreceptor population. The rods are of a single type with long, narrow outer segments and are tightly packed. The wavelength of maximum absorbance ( &lgr; max) of the visual pigment in the rods is in the region of 494 nm. Two distinct types of cone are present. The most common cone, with a stout but stubby outer segment, contains a visual pigment with &lgr; max at approximately 551 nm. A relatively rare cone, with a long, slender outer segment, contains an ultraviolet-sensitive visual pigment with &lgr; max at approximately 360 nm. All the visual pigments have chromophores based on vitamin A1. The results are discussed in relation to the behavior of P. regius. PMID- 10377273 TI - Flight speed and body mass of nectar-feeding bats (Glossophaginae) during foraging. AB - Aerodynamic theory predicts that minimum power (Vmp) and maximum range (Vmr) flight speeds increase when the body mass of an individual animal increases. To evaluate whether foraging bats regulate their flight speed within a fixed speed category relative to Vmp or Vmr, I investigated how the natural daily changes in body mass caused by feeding affected the flight speed of neotropical nectar feeding bats (Phyllostomidae: Glossophaginae) within a strictly defined, stereotyped behavioural context. Individual bats were maintained in a flight tunnel (lengths of five different types 14-50 m) with a fully automated feeding, weighing (using an electronic balance at the roost) and flight speed measuring system. Flight speeds were measured during normal nocturnal foraging activity by an undisturbed bat while it flew between the two ends of the flight tunnel to obtain food from two computer-controlled nectar-feeders. For a comparison of flight enclosure measurements with field data, flight speeds were also obtained from unrestrained bats foraging in their natural environment (Costa Rica). Foraging flight speeds spanned a range of at least a factor 3 within a single species, which demonstrates the wide range of speeds possible to these animals. Significant, positive correlations between flight speed and the natural individual variability in body mass were found in nearly all cases, with body mass exponents ranging between 0.44 and 2.1. Bats flying at normal speeds were therefore not near their upper limit of muscle power. The most reliable measurements of speed increase with mass (with individual mass changes of up to 30%) were close to the increase theoretically predicted for Vmp and Vmr for an individual bat (with constant wing span and area), which should vary as M0.42, where M is mass. This provides evidence that the glossophagine bats attemped to maintain their flight speed within a fixed speed category relative to Vmp or Vmr during foraging. Among differently sized species of glossophagine bat (N=4), flight speeds V varied with V=20M0.23, in agreement with the mass exponent of 0.21 expected from aerodynamic models for interspecific variation. In addition to the mass effect, at least five other variables significantly influenced flight speed. (1) Both mean and maximum flight speeds increased with the length and the cross-sectional area of the flight tunnel. Mean (maximum) flight speeds of 11-12 g Glossophaga soricina bats (in m s-1) were 4.6 (5.3) over a 7 m and 7. 3 (10.5) over a 50 m flight path. (2) The flight speed range adopted by a bat during one night could vary significantly between nights, independently of body mass and the effect of the size of the flight enclosure. (3) Bats flew significantly faster under illumination than in darkness. This effect was shown (i) by bats kept under natural ambient illumination that initiated foraging during the twilight phase of the evening, (ii) when bats continued to feed into the light phase directly after the dark-light transition in the laboratory and (iii) during foraging under constant, artificial illumination. (4) After a period of rest, the initial flight speed during a foraging bout was significantly increased by 25%, but declined to the mean level within 20 s of activity. (5) Flight speed could differ significantly between foraging (flight from feeder to feeder) versus non-foraging (flight from end to end of the enclosure without visiting the feeders) flights. The results of this study demonstrate a clear ability of bats to regulate their flight speed in response to small natural changes in body mass as predicted by aerodynamic theory for Vmp and Vmr. The set point in flight speed regulation, however, was influenced by multiple additional variables. PMID- 10377275 TI - Voltage-activated whole-cell K+ currents in lamina cells of the desert locust schistocerca gregaria AB - Voltage-dependent outward currents were studied in freshly dissociated somata of locust lamina cells. These currents were recorded in 142 somata using the whole cell patch-clamp technique. By measuring the reversal potential at altered external [K+] and by replacing internal K+ with Cs+, we determined that the outward currents were carried by K+. The outward currents consist of a transient A-type K+ current (KA) and a delayed-rectifier-like K+ current (KD). Amongst the cells studied, we observed two distinct groups of cells. The most obvious difference between the two groups is that in group I cells the total outward current is dominated by KA (KA/KD=12.5), whereas in group II cells KA makes a smaller contribution (KA/KD=2.1). Furthermore, in cells of group I, the KA current shows a steeper voltage-dependence of activation, where VG50 is -29.9 mV and s is 11.9 (N=22), and inactivation, where VI50 is -84.5 mV and s is -6.3 (N=18), compared with the KA current in cells of group II: VG50=-7.9 mV; s=26.6 (N=36) and VI50=-68.4 mV; s=-7.5 (N=21) (VG50 is the voltage at which the whole cell conductance G is half-maximally activated, VI50 is the voltage of half maximal inactivation and s is the slope of the voltage-dependence). The transient KA current in group I cells decayed mono-exponentially. The decay of the KA current in group II cells was fitted with a double-exponential curve and was significantly faster than in group I cells. In contrast to the large differences in KA currents, the KD currents appeared to be quite similar in the two groups of cells. PMID- 10377276 TI - Chemolithoheterotrophy in a metazoan tissue: sulfide supports cellular work in ciliated mussel gills AB - Hydrogen sulfide, a common constituent of marine intertidal sediments, is both a potent toxin of aerobic cellular respiration and an electron-rich molecule used by some prokaryotic organisms as a source of energy. In ciliated gills from Geukensia demissa, a marine mussel from sulfide-rich sediments, sulfide oxidation supports cellular work. Evidence for this comes from measurements of ciliary beat frequency (fCB) as a measure of ATP turnover rate, the rate of gill oxygen consumption ( m_dot O2) as a measure of ATP production rate, and mitochondrial cytochrome redox state as an indicator of the path of electron flow. Results from experiments performed in the presence and absence of the mitochondrial complex III inhibitor antimycin A to limit endogenous carbon substrate oxidation showed that exposure to sulfide stimulated oxygen consumption and ciliary beating, with cytochrome c being the dominant reduced species. These results, along with the resultant fCB/ m_dot O2 ratio, are qualitatively and quantitatively consistent with the hypothesis that electrons from sulfide oxidation support mitochondrial ATP production. We propose that Geukensia demissa gills use sulfide as a respiratory substrate when given the choice and thus function metabolically as facultative chemolithoheterotrophs. Similar conclusions could not be drawn for the ciliated gills from Mytilus edulis, a marine mussel from aerated habitats, or for the ciliated lungs from the phylogenetically distinct leopard frog Rana pipiens. PMID- 10377277 TI - Haemoglobin H+ equilibria in lamprey (Lampetra fluviatilis) and hagfish (Myxine glutinosa) AB - Agnathans, comprising lamprey and hagfish species, have been reported to be practically devoid of HCO3-/Cl- exchange across the red blood cell membrane. This suggests that the capacity of their haemoglobin (Hb) to remove H+ is essential for obtaining a high CO2-carrying capacity in the blood. Hydrogen ion titrations were performed on oxygenated and deoxygenated composite Hbs from river lamprey and from Atlantic hagfish at 15 degrees C and an ionic strength of 0.1 (0.1 mol l 1 KCl). Lamprey Hb was characterised by very low buffer values when the degree of oxygenation was constant, whereas the fixed-acid Haldane effect was large (uptake of approximately 0.9 H+ per monomer upon deoxygenation). Hagfish Hb, in contrast, had large buffer values and a moderate fixed-acid Haldane effect. In deoxygenated Hb, the low buffer values in lamprey correlated with the presence of only 1-1.5 titratable 'neutral' groups (normally histidines and &agr; -amino groups) per monomer, whereas there were 4-5 titratable 'neutral' groups per monomer in hagfish. The large differences in Hb/H+ equilibria between the two species reflect the early evolutionary divergence between lampreys and hagfish. With respect to CO2 transport, the special Hb/H+ equilibria and the high red blood cell pH in lamprey ensure a high concentration of free HCO3- inside the red cells in venous blood, which compensates for the absence of a shift of HCO3- to the plasma. The Hb/H+ equilibria in hagfish are less effective in ensuring a high CO2 carrying capacity given the virtual absence of a red blood cell HCO3-/Cl- exchange, and other adaptations may be involved. PMID- 10377278 TI - Pancreas development and diabetes. AB - The past few years have seen an increase in interest about the molecular and genetic events regulating pancreas development. Transcription factors such as Pdx1, p48 and Nkx2.2 have been shown to be essential for the proper differentiation of exocrine and endocrine tissue; however, pancreas development also involves intricate interactions between the pancreatic epithelium and its surrounding mesenchyme. Signalling factors emanating from the notochord have been shown to repress Sonic hedgehog expression in the endoderm whereas signals originating from the pancreatic mesenchyme determine the proportion of exocrine to endocrine tissue. Understanding the molecular and genetic events underlying pancreas development also opens the door for devising new therapeutic strategies against pancreatic diseases such as diabetes and cancer. PMID- 10377279 TI - Cardiac hypertrophy: sorting out the circuitry. AB - Cardiac hypertrophy is an adaptive response of the heart to a variety of intrinsic and extrinsic stimuli. The hypertrophic response, during which cardiomyocytes increase in size without undergoing cell division, initially serves to compensate for decreased cardiac output; however, prolonged hypertrophy can become detrimental, resulting in dilated cardiomyopathy and heart failure. Cardiac hypertrophy requires coupling of intracellular signal transduction systems with transcription factors that activate and maintain the hypertrophic program. Over the past year, signaling pathways involving G proteins, mitogen activated protein kinases and calcium-responsive phosphatases have emerged as critical regulators of cardiac hypertrophy. PMID- 10377280 TI - The PML nuclear bodies: actors or extras? AB - The PML and SP100-containing nuclear bodies (NBs) represent the best-studied example of a defined nuclear substructure the integrity of which is compromised in certain human diseases, including leukemia, neurodegenerative disorders and viral infection. Although recent progress has underscored the unexpectedly broad involvement of NB constituents in the control of cell growth, gene regulation and apoptosis in both pathological and normal contexts, evidence for a specific physiological activity within the NBs remains scant, thus precluding a unifying hypothesis for NB function. PMID- 10377281 TI - Molecular genetics of congenital hypothyroidism. AB - Congenital thyroid gland defects - resulting in reduced production of the hormones triiodothyronine (T3) and thyroxine (T4) - can be a consequence of either reduced or absent thyroid tissue (thyroid dysgenesis) or, less frequently, of impairment in the biochemical mechanisms responsible for hormone biosynthesis (thyroid dyshormonogenesis). Recent studies have revealed how mutations in the genes encoding either transcription factors or the thyroid stimulating hormone receptor cause, in humans or in mouse models, thyroid dysgenesis. This demonstrates, for the first time, the heritability of this condition. New genes responsible for thyroid dyshormonogenesis have also been discovered. PMID- 10377282 TI - The roles of RNA-binding proteins in spermatogenesis and male infertility. AB - RNA-binding proteins are essential for spermatogenesis: they are required in the nucleus of germ cells, for the production of specific mRNA isoforms, and in the cytoplasm - where proteins required for chromatin condensation and for changes in cell morphology are translated long after transcription ceases. Some of the RNA targets and the RNA-binding proteins themselves have been identified recently. Both nuclear and cytoplasmic proteins are affected in examples of azoospermia in men. PMID- 10377283 TI - Cellular and molecular mechanisms of memory: the LTP connection. AB - Studies of the cellular and molecular mechanisms of memory formation have focused on the role of long-lasting forms of synaptic plasticity such as long-term potentiation (LTP). A combination of genetic, electrophysiological and behavioral techniques have been used to examine the possibility that LTP is a cellular mechanism of memory storage in the mammalian brain. Although a definitive answer remains elusive, it is clear that in many cases manipulations that alter LTP alter memory, and training regimens that produce memory can produce LTP-like potentiation of synaptic transmission. PMID- 10377284 TI - Telomerase: Dr Jekyll or Mr Hyde? AB - The past year has seen the ectopic expression of human telomerase and the consequent increased replicative lifespan of cells, whereas mice lacking telomerase have lived and reproduced for six generations. Core telomerase activity from various organisms was reconstituted in vitro, yet how its action is regulated remains largely unknown. Telomerase activation preceded oncogenic transformation in some human cell types, yet was lacking in other transformed cells. These advances highlight the potentials of telomerase-based therapeutics and warn of their pitfalls. PMID- 10377285 TI - Pain. AB - Advances in our understanding of the activation of peripheral damage-sensing neurons (nociceptors) over the past year have been complemented by electrophysiological and imaging studies of central nervous system pain-related centres. The manipulation of gene expression in a reversible and cell type specific way combined with imaging and electrophysiological studies holds promise for helping us to identify the spatial and molecular substrates of pain perception with increasing precision and gives hope for improved analgesic therapies. PMID- 10377286 TI - Advances in the molecular genetics of obesity. AB - The past year has seen substantial progress in our understanding the molecular mechanisms of bodyweight regulation, particularly in the central and peripheral actions of the leptin and melanocortin signaling pathways (e.g. leptin stimulation of angiogenesis and suppression of cytokine production). Important advances also include the identification of mutations in components of the leptin and melanocortin pathways in human obese families. Expanding from the positional cloning of leptin some five years ago, the mouse continues to be a central focus of study, particularly the way in which different bodyweight-sensing pathways interact in different feeding states. PMID- 10377287 TI - Genes involved in deafness. AB - Remarkable progress has been made over the past few years in the field of hereditary deafness. To date, mutations in at least 35 genes are known to cause hearing loss. We are now beginning to understand the function of many of these genes, which affect diverse aspects of ear development and function. PMID- 10377288 TI - Transcription factors for lens development assessed in vivo. AB - Lens-cell differentiation occurs at a fairly early stage of embryogenesis and results in very simple tissue architecture. These features allow the embryonic lens to provide a paradigm of tissue development starting from tissue induction to tissue maturation. Not only have a number of transcription factors participating in lens development been identified but their actual functions are now assessed by modern approaches utilizing genetic and tissue manipulations of embryos. PMID- 10377289 TI - The trithorax-group and polycomb-group chromatin modifiers: implications for disease. AB - The chromatin structure associated transcriptional memory function by Polycomb group and trithorax-group protein complexes integrate normal development with control of cellular proliferation. This is illustrated by recent mechanistic insights that link Polycomb repression with histone deacetylases and the identification of new target genes that provide a connection to cell cycle control and tumorigenesis. PMID- 10377290 TI - Muscular dystrophies: alterations in a limited number of cellular pathways? AB - Identification of new genes involved in muscle disorders has dramatically changed the traditional clinical classification of the large and heterogeneous group of the muscular dystrophies. Results obtained in recent years by positional candidate cloning have demonstrated the role of the sarcolemma and of the nuclear envelope in normal muscle function and have elucidated molecular pathways perturbed by mutations that lead to muscular dystrophy. PMID- 10377291 TI - Odorant receptor genes in humans. AB - The sense of smell is highly sophisticated in vertebrates but Homo sapiens ranks low in olfactory performance when compared to other species - why? Olfaction initiates with the interaction of odorants with specific receptors on the surface of olfactory sensory neurons in the nose. The genes encoding odorant receptors form the largest family in the vertebrate genome, numbering as many as 1000 in rodents. It has recently come to light that the repertoire of human odorant receptor genes, unlike in other vertebrates, is riddled with pseudogenes. PMID- 10377292 TI - Molecular biology of prion propagation. AB - The occurrence of new variant Creutzfeldt-Jakob disease and the experimental confirmation that it is caused by the same prion strain as BSE has dramatically highlighted the need for a precise understanding of the molecular basis of prion propagation. The molecular basis of prion-strain diversity, previously a major challenge to the protein-only model, is now becoming clearer. The conformational change thought to be central to prion propagation, from a predominantly alpha helical fold to one predominantly comprising beta-structure, can now be reproduced in vitro, and the ability of beta-PrP to form fibrillar aggregates provides a plausible molecular mechanism for prion propagation. These and other advances in the fundamental biology of prion propagation are leading to prion diseases becoming arguably the best understood of the neurodegenerative conditions and strategies for the development of rational therapeutics are becoming clearer. PMID- 10377293 TI - Development of the mammalian enteric nervous system. AB - The mammalian enteric nervous system is derived from neural crest cells which invade the foregut and hindgut mesenchyme. It has been established that signalling molecules produced by the mesenchyme of the gut wall play a critical role in the development of the mammalian enteric nervous system. Recent studies have characterised further the role of such molecules and have identified novel extracellular and intracellular signals that are critical for enteric ganglia formation. PMID- 10377294 TI - Misuse of "coronary heart disease". PMID- 10377295 TI - Flying after heart surgery. PMID- 10377297 TI - The difficulties in assessing patients with moderate aortic stenosis. PMID- 10377298 TI - Exercise testing to guide surgery in aortic stenosis. PMID- 10377299 TI - Is aortic valve resistance more clinically meaningful than valve area in aortic stenosis? PMID- 10377300 TI - The relation between transaortic pressure difference and flow during dobutamine stress echocardiography in patients with aortic stenosis. AB - OBJECTIVE: To investigate the relation between transaortic pressure difference and flow in patients with aortic stenosis. METHODS: 50 asymptomatic patients with all grades of aortic stenosis were studied using dobutamine stress echocardiography. Individual plots of mean pressure drop against flow were drawn. Comparisons were made between grades of aortic stenosis as defined by the continuity equation. RESULTS: A significant linear relation between pressure difference and flow was found in 34 patients (68%). There was a significant curvilinear relation in four (8%), while no significant regression line could be fitted in 12 (24%). In the 34 patients with linear fits, the slopes (mean (SD)) were 0.08 (0.07) in mild, 0.10 (0.04) in moderate, and 0.22 (0.16) in severe aortic stenosis (p = 0. 0055). CONCLUSIONS: Transaortic pressure difference can be related directly to flow in many patients with all grades of aortic stenosis. However, there are individual differences in slope and intercept suggesting that resistance calculated at rest may not always be representative. Raw pressure drop/flow plots may be an alternative method of describing valve function. PMID- 10377301 TI - QT dispersion is reduced after valve replacement in patients with aortic stenosis. AB - OBJECTIVE: To investigate whether QT dispersion is a reliable index of the severity of aortic stenosis and left ventricular hypertrophy in the setting of aortic stenosis. DESIGN: A retrospective analysis of the results of echocardiography and electrocardiography before and after aortic valve replacement. SETTING: Tertiary centre. PATIENTS: 36 men (30 white and six black) with symptomatic aortic stenosis requiring valve replacement. RESULTS: All patients had significant aortic stenosis (mean (SD) aortic valve area 0.68 (0.18) cm2) and evidence of left ventricular hypertrophy (left ventricular mass index (LVMI): 267 (90) g/m2). Before aortic valve replacement, QT dispersion was correlated with mean aortic valve area and LVMI (r = 0.697, p < 0.001, and r = 0.59, p < 2.4 x 10(-6), respectively). QT dispersion and QT corrected for heart rate dispersion decreased from 133 (54) to 71 (33) ms and from 151 (64) to 94 (76) ms, respectively (p < 0.001 for both). LVMI regressed after aortic valve replacement to 190 (79) g/m2, p < 0.01. CONCLUSIONS: QT dispersion is increased in association with LVMI in patients with significant symptomatic aortic stenosis. Aortic valve replacement reduces QT dispersion and LVMI. QT dispersion could be a useful indicator of risk and risk reduction in patients with significant symptomatic aortic stenosis. PMID- 10377302 TI - Aortic root dilatation in young men with normally functioning bicuspid aortic valves. AB - OBJECTIVE: To evaluate the dimensions of the aortic root in a selected population of young males with isolated normally functioning bicuspid aortic valve. DESIGN AND SETTING: Echocardiographic and Doppler evaluation of conscripts with bicuspid aortic valve at the time of military pre-enrolment screening in two military hospitals. SUBJECTS AND METHODS: 66 consecutive young men with a normally functioning bicuspid aortic valve were studied to assess aortic size at four aortic levels: annulus, sinuses of Valsalva, supra-aortic ridge, and proximal ascending aorta; 70 consecutive normal young subjects, matched for age and body surface area, were used as controls. RESULTS: In men with a bicuspid aortic valve, the diameter of the aortic root was significantly larger than in controls at the sinuses (3.16 (0.37) v 2.87 (0.31) cm, p < 0.001), at the supra-aortic ridge (2.64 (0.46) v 2.47 (0.28) cm, p = 0.01), and at the level of the proximal ascending aorta (3.12 (0.48) v 2.69 (0.28) cm, p < 0.001). The prevalence of aortic root dilatation was 7.5% at the annulus (5/66), 19.6% at the sinuses (13/66), 15% at the supra-aortic ridge (10/66), and 43.9% at the ascending aorta (29/66); 32 subjects (48%) had aortic root dimensions comparable with controls, while 34 (52%) had definitely abnormal aortic root dimensions. CONCLUSIONS: Aortic root enlargement in people with a bicuspid aortic valve occurs independently of haemodynamic abnormalities, age, and body size. However, there appear to be different subgroups of young adults with bicuspid aortic valves, one of which is characterised by aortic dilatation, possibly caused by a congenital abnormality of the aortic wall. PMID- 10377303 TI - Anticoagulation in pregnant women with mechanical heart valve prostheses. AB - OBJECTIVE: To evaluate the outcome of pregnancy in women with mechanical heart valve prostheses in relation to the anticoagulant treatment used in the first trimester and the incidence of thrombotic and bleeding complications. METHODS: 92 pregnancies in 59 women were followed between 1986 and 1997. In 31 pregnancies, oral anticoagulants were discontinued when pregnancy was diagnosed and subcutaneous heparin was started (12 500 U every 12 hours) adjusted to prolong the adjusted partial thromboplastin time to twice the control level. In the second trimester oral anticoagulants were resumed but changed to heparin again 15 days before the expected delivery date. In 61 pregnancies oral anticoagulants were continued during the first trimester. The same regimen of heparin was used for delivery. RESULTS: Abortion or fetal losses were similar (p = 0. 5717) in women exposed to oral anticoagulants in the first trimester (13/61; 25%) compared with those who received adjusted subcutaneous heparin (6/31; 19%). Embolic episodes were more common (p = 0.0029) in women who received heparin (4.92%) compared with those on oral anticoagulants (0.33%). Embolic episodes were cerebral and transient. No valve thromboses were observed. No malformations appeared in the 71 newborns, except for one case of hydrocephalus. There were no maternal deaths secondary to thrombotic complications. The only death was the result of major bleeding after the delivery of a premature stillborn. CONCLUSIONS: Oral anticoagulants seem to be safer for the mother than adjusted subcutaneous heparin. Heparin does not offer a clear advantage over oral anticoagulation in the pregnancy outcome. PMID- 10377304 TI - Use of self expanding stents in stenotic aortopulmonary shunts in adults with complex cyanotic heart disease. AB - OBJECTIVE: To describe the use of self expanding stents in treating long segment stenosis of aortopulmonary shunts (APS) in adults. DESIGN: Clinical records, catheterisation data, cineangiograms, and operation notes of four consecutive patients undergoing stent implantation since December 1994 were studied retrospectively. SETTING: A tertiary referral centre for cardiac disease. SUBJECTS: Four patients underwent cardiac catheterisation because of clinical deterioration. Their age ranged between 23 and 32 years. The underlying diagnosis was complex cyanotic heart disease in all. Three had a stenotic interposition graft, and one had a classic Blalock shunt. RESULTS: There was one technical failure owing to migration of the stent distal to an ostial stenosis. The ability index, resting oxygen saturation, and exercise tolerance improved in the remainder. Their medium term results have been excellent. CONCLUSIONS: This technique may further palliate adult patients with complex congenital heart disease, though the long term patency of stents is unknown. PMID- 10377305 TI - Incidence of secondary pulmonary hypertension in adults with atrial septal or sinus venosus defects. AB - OBJECTIVE: To examine the incidence of raised pulmonary artery pressure and resistance in adults with isolated atrial septal defect within the oval fossa (so called secundum defect) or sinus venosus defect. DESIGN: A historical, retrospective, unrandomised study. SETTING: A tertiary referral centre. METHODS: Cardiac catheterisation was performed in all patients, with measurement of pulmonary artery pressure and resistance. Pulmonary to systemic flow ratio was calculated using the Fick principle. Pulmonary hypertension was defined as mean pulmonary artery pressure > 30 mm Hg, and increased resistance as an Rp/Rs ratio > 0.3. PATIENTS: All patients with a secundum atrial septal or sinus venosus defect who presented between July 1988 and December 1997 were enrolled in the study. RESULTS: Pulmonary artery pressure and resistance in the patients with sinus venosus defect (n = 31) was higher than in patients with atrial septal defect (n = 138). Pulmonary hypertension was present in 26% of patients with sinus venosus and in 9% of patients with atrial septal defect. The incidence of raised pulmonary vascular resistance was 16% in patients with sinus venosus and 4% in patients with atrial septal defect. The increase in resistance occurred at a younger age in sinus venosus defect than in atrial septal defect. CONCLUSIONS: Patients with sinus venosus defect have higher pulmonary pressures and resistances and develop these complications at younger age than patients with atrial septal defects. Thus they should be managed differently than patients with "simple" atrial septal defects. PMID- 10377306 TI - Spectrum of congenital heart defects and extracardiac malformations associated with chromosomal abnormalities: results of a seven year necropsy study. AB - OBJECTIVE: To analyse the spectrum of congenital heart malformations, the frequency of extracardiac malformations, and the proportion of chromosome aberrations among fetuses sent for necropsy. MATERIAL: Necropsies were performed on 815 fetuses-448 induced abortions (55%), 220 spontaneous abortions (27%), and 147 stillbirths (18%)-during a seven year period (1991-97) in the department of pathology of the Charite Medical Centre in Berlin. A congenital heart defect was identified in 129 cases (16%). For all 129 fetuses, karyotyping and an ultrasound examination had been performed. RESULTS: Congenital heart defects were present in 22% of induced abortions (99 cases), 9% of spontaneous abortions (20 cases), and 7% of stillbirths (10 cases). The heart malformations were classified into 13 categories. A fetus with more than one defect was included only in the category of the most serious defect. The malformations in order of frequency were: ventricular septal defect (VSD) (28%), atrioventricular septal defect (AVSD) (16%), hypoplastic left heart (HLH) (16%), double outlet right ventricle (DORV) (12%), coarctation of the aorta (CoA) (6%), transposition of the great arteries (TGA) (4%), aortic valve stenosis (AoVS) (4%), tetralogy of Fallot (TOF) (3%), truncus arteriosus communis (TAC) (3%), pulmonary valve stenosis/pulmonary valve atresia (PaVS/PaVA) (3%), tricuspid atresia (TA) (3%), single ventricle (SV) (1.5%), and atrial septal defect (ASD) (0.5%). The most common congenital heart defects were VSD, AVSD, HLH, and DORV, which made up 72% of all the cases. In 11 cases the heart defect was isolated (no other cardiovascular or extracardiac malformations present), 85 cases (66%) were associated with additional cardiac malformations, 85 cases (66%) were associated with extracardiac malformations, and chromosome anomalies were detected in 43 cases (33%). CONCLUSIONS: Fetal congenital heart malformations are common. These defects are often associated with other cardiovascular and extracardiac malformations, as well as with chromosome anomalies. Complex heart defects such as AVSD, HLH, and DORV are frequent in fetuses, as they often lead to spontaneous abortion or stillbirth or, after prenatal diagnosis, to deliberate termination of pregnancy. PMID- 10377307 TI - Abnormalities in liver function and coagulation profile following the Fontan procedure. AB - OBJECTIVE: To investigate liver function and coagulation disorders in patients with a Fontan circulation at different time intervals after surgery. DESIGN: Retrospective analysis of clinical data and cross sectional study relating liver function and coagulation profile to time since surgery, in 28 surviving patients after the modified Fontan procedure. PATIENTS: 20 patients (71%) with atriopulmonary anastomosis, seven (25%) with atrioventricular anastomosis, and one (4%) with total cavopulmonary connection. Follow up ranged from 2.0 to 21.8 years (mean 11.1). RESULTS: Abnormal liver function tests, mainly reflecting cholestasis, were present in 21 patients who had a significantly longer follow up (p < 0.01). Protein synthesis was normal in almost all patients. Coagulation profile showed abnormalities in 22 patients. "Procoagulant" abnormalities-that is, decreased plasminogen and protein C activity-were found in 11 and five patients, respectively. The extent of these abnormalities was less in patients with a longer follow up. Anticoagulant abnormalities were factor V deficiency in 16 patients and factor VII deficiency in 17, resulting in a prolonged prothrombin time in 19 patients. Thirteen patients had both pro- and anticoagulant abnormalities. A prethrombotic state was present in five patients, with a significantly longer mean time interval since surgery (p = 0.05). Thus, although the individual procoagulant indices decreased with increasing time intervals since surgery, a prethrombotic state was found particularly in patients with a long term follow up. CONCLUSIONS: Mild cholestasis was mainly present in Fontan patients with a long duration of follow up. Along with laboratory procoagulant abnormalities indicating a prethrombotic state, anticoagulant abnormalities were also present. The coagulation profile varied at different time intervals after surgery. Thus detailed evaluation should be performed regularly, and the use of anticoagulants should be considered in every patient. Long term prospective studies are needed to evaluate the individual fluctuations of coagulation profile over time following a Fontan procedure. PMID- 10377308 TI - Ten year survival after heart transplantation: palliative procedure or successful long term treatment? AB - OBJECTIVE: To investigate the long term outcome and prognostic factors after heart transplantation. SETTING: University hospital. SUBJECTS: 120 heart transplant patients (98 male, 22 female; underlying disease: dilated cardiomyopathy in 69, coronary artery disease in 42, miscellaneous in nine) who had undergone heart transplantation between October 1984 and October 1987. Immunosuppressive treatment was comparable in all patients and rejection episodes were treated in a uniform manner. METHODS: Functional status, quality of life, and potential predictors for long term survival were investigated. RESULTS: Actuarial survival rates were 65% at five years and 48% at 10 years; 58 patients survived > 10 years. The major causes of death were cardiac allograft vasculopathy (39%), acute rejection (18%), infection (11%), and malignancy (11%). Long term survivors had good exercise tolerance assessed by the New York Heart Association classification: 47 (81%) in grade I/II; 11 (19%) in grade III/IV. Echocardiography showed good left ventricular function in 48 patients. On angiography, severe allograft vasculopathy was present in only 16 patients (28%). Renal function was only slightly impaired, with mean (SD) serum creatinine of 148.5 (84.9) micromol/l. Multiple potential predictors of long term survival were analysed but none was found useful. CONCLUSIONS: Heart transplantation represents a valuable form of treatment. Survival for more than 10 years with a good exercise tolerance and acceptable side effects from immunosuppression can be achieved in about 50% of patients. PMID- 10377309 TI - Cardiovascular disease in the Netherlands, 1975 to 1995: decline in mortality, but increasing numbers of patients with chronic conditions. AB - OBJECTIVE: To examine the relation between trends over time in mortality and hospital morbidity caused by various cardiovascular diseases in the Netherlands. DESIGN: Trend analysis by Poisson regression of national data on mortality and hospital admissions from 1975 to 1995. SUBJECTS: The Dutch population. RESULTS: All cardiovascular diseases combined were responsible for 39% of all deaths and 16% of all hospital admissions in 1995. From 1975 to 1995, age adjusted cardiovascular mortality declined by an annual change of -2.0% (95% confidence intervals (CI) -2.1% to -1.9%), while in the same period age adjusted discharge rates increased annually by 1. 3% (95% CI 1.1% to 1.5%). Around 60% of the gain in life expectancy in this period was related to lower cardiovascular mortality. For mortality, major reductions were seen in coronary heart disease (annual change -2.9%) and in stroke (-2.1%), whereas the increase in hospital admissions was mainly caused by chronic manifestations of coronary heart disease (5.1%), heart failure (2.1%), and diseases of the arteries (1.8%). In recent years, the gap between men and women at risk of dying from coronary heart disease became smaller for those aged 10 pg/ml were seen in 15% of patients, and aldosterone concentrations > 144 pg/ml were seen in 38% of patients. AII concentrations were significantly correlated (p < 0.001) with ACE but not with aldosterone concentrations. Aldosterone concentrations were not significantly correlated with ACE concentrations. CONCLUSIONS: AII "reactivation" occurred in 15% and failure of aldosterone suppression in 38% of routine CHF patients taking ACE inhibitor treatment. AII "reactivation" was associated with both low and high levels of ACE activity, which suggests that multiple different mechanisms are at play. In patients with high plasma ACE concentrations, non-compliance should be considered along with inadequate dose titration. In patients with low plasma ACE and high AII concentrations, non-ACE mediated production of AII may be operative. Raised aldosterone concentrations appear to be more common than AII "reactivation". It is important to establish the cause of detectable or increased AII concentrations in a heart failure patient treated with an ACE inhibitor. The measurement of serum ACE may help to identify the likely cause as poor compliance or inadequate dose. PMID- 10377311 TI - Combined assessment of reflow and collateral blood flow by myocardial contrast echocardiography after acute reperfused myocardial infarction. AB - OBJECTIVE: To evaluate the combined assessment of reflow and collateral blood flow by myocardial contrast echocardiography after myocardial infarction. DESIGN: Myocardial contrast echocardiography was performed in patients with acute myocardial infarction shortly after successful coronary reperfusion (TIMI 3 patency) by direct angioplasty. Collateral flow was assessed before coronary angioplasty, and contrast reflow was evaluated 15 minutes after reperfusion. The presence of contractile reserve was assessed by low dose dobutamine echocardiography (5 to 15 micrograms/kg/min) at (mean (SD)) 3 (2) days after myocardial infarction. Recovery of segmental function (myocardial viability) was evaluated by resting echocardiography at a two month follow up. The study was prospective. PATIENTS: 35 consecutive patients referred for acute transmural myocardial infarction. RESULTS: Contrast reflow was observed in 20 patients (57%) and collateral flow in 14 (40%). Contrast reflow and collateral contrast flow were both correlated with reversible dysfunction on initial dobutamine echocardiography and at follow up (p < 0.05). The presence of reflow or collateral flow on myocardial contrast echocardiography was a highly sensitive (100%) but weakly specific (60%) indicator of segmental dysfunction recovery. Simultaneous presence of contrast reflow and collateral flow was more specific of reversible dysfunction than reflow alone (90% v 60%). CONCLUSIONS: Combined assessment of reflow and collateral blood flow enhanced the sensitivity of myocardial contrast echocardiography in predicting myocardial viability after acute, reperfused myocardial infarction. The simultaneous presence of reflow and collateral blood flow was highly specific of recovery of segmental dysfunction. PMID- 10377312 TI - Arterial remodelling of native human coronary arteries in patients with unstable angina pectoris: a prospective intravascular ultrasound study. AB - OBJECTIVE: To investigate the use of intravascular ultrasound (IVUS) in detecting the presence of arterial remodelling in patients with unstable angina. DESIGN: Prospective case study. PATIENTS: 60 of 95 consecutively admitted patients with unstable angina (41 male, 19 female), mean (SD) age 61.2 (8.1) years. INTERVENTIONS: Qualitative and quantitative coronary angiography and IVUS. MAIN OUTCOME MEASURES: Adaptive or constrictive remodelling (AR, CR) was considered present when the cross sectional area of the external elastic membrane at the lesion site was larger than the proximal cross sectional area or smaller than the distal cross sectional area, respectively. RESULTS: 22 of the 60 patients (37%) showed AR and 14 (23%) showed CR. No remodelling was seen in 24 patients (group NR). The plaque contained more thrombus and plaque rupture in group AR than in groups CR and NR (thrombus: 91% v 50% and 67%, respectively, p = 0.023; rupture: 73% v 29% and 42%, p = 0.020). AR was associated with a larger plaque cross sectional area (12.6 (SD 4.6) mm2 v 10.8 (6.3) and 9.2 (3.7) mm2, p = 0.001) and larger external elastic membrane cross sectional area (16.5 (5.8) mm2 v 13.2 (5.2) and 14.4 (3.6) mm2, p = 0.01 in group AR v groups CR and NR, respectively), while the plaque burden was larger in groups AR (74.9 (9.1)%) and CR (72.4 (16.6)%) than in group NR (66.2 (18.1)%, p = 0.005). CONCLUSIONS: IVUS is capable of detecting adaptive and constrictive remodelling of target lesions and its relation to plaque morphology in unstable angina. PMID- 10377313 TI - Alterations of autonomic nervous activity in recurrence of variant angina. AB - OBJECTIVE: To investigate whether autonomic nervous activity is involved in the recurrence of spontaneous coronary spasm in variant angina. DESIGN: Retrospective analysis. SETTING: Cardiology department of a university hospital. PATIENTS: 18 patients with variant angina were divided into single attack group (SA; nine patients) and multiple attack group (MA; nine patients) according to the frequency of ischaemic episodes with ST segment elevation during 24 hour Holter monitoring. METHODS: Heart rate variability indices were calculated using MemCalc method, which is a combination of the maximum entropy method for spectral analysis and the non-linear least squares method for fitting analysis, at 30 second intervals for 30 second periods, from 40 minutes before the attack to 30 minutes after the attack. High frequency (HF; 0.04-0.15 Hz) was defined as a marker of parasympathetic activity, and the ratio of low frequency (LF; 0.15-0.40 Hz) to high frequency (LF/HF) as an indicator of sympathetic activity. The averaged value during the 40 to 30 minute period before an attack was defined as the baseline. RESULTS: Compared with baseline, the HF component decreased in both groups at two minutes before the attack (p < 0.01), and the LF/HF ratio decreased at three minutes before the attack (p < 0.01). The baseline LF/HF was lower in the MA group than in the SA group (p < 0. 01). CONCLUSIONS: A reduction of sympathetic activity may play a key role in determining the recurrence of transient ischaemic events caused by spontaneous coronary spasm in patients with variant angina. PMID- 10377314 TI - Clinical outcome of patients treated with spinal cord stimulation for therapeutically refractory angina pectoris. The Working Group on Neurocardiology. AB - OBJECTIVE: To determine morbidity and mortality characteristics in patients treated with electrical neuromodulation for refractory angina pectoris. DESIGN: A retrospective multicentre study of patients treated with spinal cord stimulation between 1987 and 1997; 21 centres were contacted and 14 responded. SETTING: Specialist centres worldwide. PATIENTS: Questionnaires were returned on 517 patients, of whom 71% were male. One was lost to follow up. Mean (SD) age was 63.9 (10.1) years. Duration of angina pectoris was 8.1 (6.3) years. RESULTS: Before spinal cord stimulation, 66% of the patients had experienced myocardial infarction, 68% had three vessel disease, and in 24% the left ventricular ejection fraction (LVEF) was /= 71 years were independent predictors of mortality. During spinal cord stimulation, New York Heart Association functional class improved from 3.5 to 2.1 (p < 0.01); 25 of the deceased patients (24%) and 32 survivors (8%) experienced myocardial infarction; hospital admissions were significantly (p < 0.001) more common in the deceased group (66% v 37%). CONCLUSIONS: The clinical outcome of patients with intractable angina is not adversely affected by the chronic use of neurostimulation. PMID- 10377315 TI - Images in cardiology: Mycotic aneurysm of the left pulmonary artery in a child with tetralogy of Fallot and Streptococcus viridans infective endocarditis. PMID- 10377316 TI - Spinal cord stimulation significantly decreases the need for acute hospital admission for chest pain in patients with refractory angina pectoris. AB - OBJECTIVE: To assess the impact of spinal cord stimulation (SCS) on the need for acute admissions for chest pain in patients with refractory angina pectoris. DESIGN: Retrospective analysis of case records. PATIENTS: 19 consecutive patients implanted for SCS between 1987 and 1997. All had three vessel coronary disease, and all were in New York Heart Association functional group III/IV. METHODS: Admission rates were calculated for three separate periods: (1) from initial presentation up until last revascularisation; (2) from last revascularisation until SCS implantation; (3) from SCS implantation until the study date. Post revascularisation rates were then compared with post-SCS rates, without including admissions before revascularisation, as this would bias against revascularisation procedures. RESULTS: Annual admission rate after revascularisation was 0.97/patient/year, compared with 0.27 after SCS (p = 0.02). Mean time in hospital/patient/year after revascularisation was 8.3 days v 2.5 days after SCS (p = 0.04). No unexplained new ECG changes were observed during follow up and patients presented with unstable angina and acute myocardial infarction in the usual way. CONCLUSIONS: SCS is effective in preventing hospital admissions in patients with refractory angina, without masking serious ischaemic symptoms or leading to silent infarction. PMID- 10377317 TI - Handgrip exercise increases postocclusion hyperaemic brachial artery dilatation. AB - OBJECTIVE: To examine the effect of handgrip exercise induced ischaemia on non invasive assessment of endothelial function in the brachial artery. DESIGN AND SETTING: High frequency ultrasound was used to measure brachial artery diameter at rest and after reactive hyperaemia induced by forearm cuff occlusion with and without handgrip exercise induced ischaemia. SUBJECTS: 10 healthy subjects, < 40 years, without known cardiovascular risk factors. MAIN OUTCOME MEASURES: Brachial artery dilatation and blood flow. RESULTS: Hyperaemia following forearm occlusion with handgrip exercise induced ischaemia increased brachial artery diameter significantly more than hyperaemia following occlusion alone, 6.9 (3.2)% and 4.5 (1.6)%, respectively (95% confidence interval 0.3% to 4.5%). There was no difference in peak blood flow with and without exercise induced ischaemia CONCLUSIONS: Handgrip exercise induced ischaemia with forearm occlusion caused more pronounced brachial artery dilatation than occlusion alone without change in peak blood flow. This suggests continued brachial artery responsiveness to the stimulus of ischaemia despite maximum blood flow and peripheral vasodilatation with occlusion alone. PMID- 10377318 TI - Stabilisation of medically refractory ventricular arrhythmia by intra-aortic balloon counterpulsation. AB - OBJECTIVE: To review the efficacy of intra-aortic balloon counterpulsation (IABCP) in medically refractory ventricular arrhythmia. DESIGN: Retrospective analysis of the outcome of patients with ventricular arrhythmia treated with IABCP after transfer between 1992 and 1997. SETTING: Tertiary cardiac referral centre. PATIENTS: 21 patients (mean age 58 years) who underwent IABCP for control of ventricular arrhythmia. All had significant left ventricular impairment (mean ejection fraction 28.6%); 18 had coronary artery disease. RESULTS: Before IABCP, 10 patients had incessant monomorphic ventricular tachycardia and 11 had paroxysmal ventricular tachycardia and/or ventricular fibrillation (VT/VF). IABCP resulted in suppression of ventricular arrhythmia in 18 patients, of whom 13 were weaned from IABCP. After stabilisation of ventricular arrhythmia, 10 patients were maintained on medical treatment alone and one underwent endocardial resection. IABCP was maintained until cardiac transplantation in five patients. One patient had a fatal arrest before discharge and one died from progressive heart failure. IABCP failed to control ventricular arrhythmia in three patients and was subsequently discontinued. A cardiac assist device was employed in one of these until cardiac transplantation; the other two were eventually stabilised on medical treatment. Nineteen patients were discharged from hospital. Overall survival was 95% at mean follow up of 25.7 months. CONCLUSIONS: IABCP can be an effective means of controlling refractory ventricular arrhythmia, allowing time for the institution of more definitive treatment. PMID- 10377319 TI - Images in cardiology: Candida endocarditis of the right heart. PMID- 10377320 TI - Initial experience in the extraction of chronically implanted pacemaker leads using the Excimer laser sheath. AB - OBJECTIVE: To assess the safety and efficiency of the Excimer laser sheath in extracting chronically implanted pacemaker leads. PATIENTS: Eight patients were studied (one female, mean age 62 years, range 34 to 77) with 17 pacemaker leads (five atrial, 10 ventricular, two implantable defibrillator). The mean implantation time was 65 months (range 23 to 188). The indications for lead extraction were chronic infection (7), superior vena cava obstruction (4), lead malfunction (4), and pain (2). METHODS: A prospective analysis of the use of the Excimer laser sheath in extracting chronically implanted pacemaker leads. Laser sheath extraction was undertaken if conventional extraction techniques with simple traction or traction with a locking stylet had failed. If laser sheath extraction was unsuccessful, basket retrieval of the lead from the groin was performed. RESULTS: Complete lead removal was achieved in 16 leads (94%). In one case the electrode tip was left behind without complication. Extraction was achieved with the laser sheath alone in 16 leads. Basket retrieval was required in one case after laser failure. There were no complications. CONCLUSIONS: The Excimer laser sheath appears to be an effective and safe technique for extracting chronically implanted pacemaker leads. It can be used in combination with the currently available techniques for successful lead extraction. PMID- 10377321 TI - Images in cardiology: Apparent fracture of a pacemaker lead. PMID- 10377322 TI - Cardiac involvement in Emery Dreifuss muscular dystrophy: a case series. AB - Three patients with Emery Dreifuss muscular dystrophy are reported. Emery Dreifuss muscular dystrophy is an X linked muscular dystrophy, in which locomotor involvement is characteristically mild and slowly progressive. The effect on the heart becomes apparent in the teenage years and is characterised by cardiac conduction defects and infiltration of the myocardium by fibrous and adipose tissue. It first affects the atria, which results in atrial paralysis; treatment with ventricular pacing is usually needed. Female carriers can develop heart problems and are at risk of sudden death. Relatives of affected patients should be offered screening with electrocardiography and echocardiography. PMID- 10377323 TI - Images in cardiology: Cutting balloon for treatment of severe peripheral pulmonary stenosis in a child. PMID- 10377324 TI - Aortic obstruction caused by device occlusion of patent arterial duct. AB - A 2 year old girl is reported in whom deployment of the Amplatzer ductal occluder caused significant aortic obstruction, requiring surgical removal of the device. This case emphasises the need for careful echocardiographic and angiographic assessment of the position of the Amplatzer ductal occluder before and after detaching the device from its delivery system, with particular emphasis on the position of the aortic retention ring. Careful assessment of ductal anatomy must guide case selection. PMID- 10377325 TI - Erosion of the left ventricle by the epicardial patch of an automatic implantable cardioverter defibrillator. AB - A 56 year old man with an implantable cardioverter defibrillator was admitted with chest pain and collapse. Erosion of the left ventricle by an epicardial patch was confirmed by thoracotomy, but surgical repair was impossible. This rare complication should be considered in patients with a history of cardioverter defibrillators implanted by thoracotomy. PMID- 10377328 TI - Cellular defects and altered gene expression in PC12 cells stably expressing mutant huntingtin. AB - Expanded polyglutamine tracts cause huntingtin and other proteins to accumulate and aggregate in neuronal nuclei. Whether the intranuclear aggregation or localization of a polyglutamine protein initiates cellular pathology remains controversial. We established stably transfected pheochromocytoma PC12 cells that express the N-terminal fragment of huntingtin containing 20 (20Q) or 150 (150Q) glutamine residues. The 150Q protein is predominantly present in the nuclei, whereas the 20Q protein is distributed throughout the cytoplasm. Electron microscopic examination confirmed that most of the 150Q protein is diffuse in the nucleus with very few microscopic aggregates observed. Compared with parental PC12 cells and cells expressing 20Q, cells expressing 150Q display abnormal morphology, lack normal neurite development, die more rapidly, and are more susceptible to apoptotic stimulation. The extent of these cellular defects in 150Q cells is correlated with the expression level of the 150Q protein. Differential display PCR and expression studies show that cells expressing 150Q have altered expression of multiple genes, including those that are important for neurite outgrowth. Our study suggests that mutant huntingtin in the nucleus is able to induce multiple cellular defects by interfering with gene expression even in the absence of aggregation. PMID- 10377329 TI - Distinct subtypes of metabotropic glutamate receptors mediate differential actions on excitability of spinal respiratory motoneurons. AB - Metabotropic glutamate receptors (mGluRs) modulate neuronal function by affecting excitability and altering synaptic transmission. We have shown that the mGluR agonist (1S,3R)-1-amino-1, 3-cyclopentanedicarboxylic acid (1S,3R-ACPD) has multiple actions on phrenic motoneurons (PMNs), including reduction of inspiratory-modulated synaptic currents and an increase of neuronal excitability. We hypothesized that these actions were mediated by different mGluR subtypes. We have now identified the involvement of mGluR subtypes and their roles in modulating the excitability of PMNs and the consequent inspiratory motor output in an in vitro neonatal rat brainstem-spinal cord preparation. Activation of postsynaptic group-I mGluRs increases PMN excitability, associated with the production of an inward current and a decrease in membrane conductance, whereas activation of group-II or group-III mGluRs decreases PMN inspiratory-modulated synaptic current, probably via a presynaptic mechanism. To confirm further the distinction and the involvement of group-I and group-II/-III receptor subtypes affecting PMN excitability, we used the membrane permeable cAMP analog 8-bromo cAMP (8-Br-cAMP) to elevate intracellular cAMP concentration to mask or occlude any effects mediated via the cAMP cascade. 8-Br-cAMP attenuated the reduction of the inspiratory-modulated activity of PMNs by both (S)-4-carboxy-3 hydroxyphenylglycine (4C3HPG) and L-(+)-2-amino-4-phosphonobutyric acid (L-AP4), agonists for group-II and group-III mGluRs, respectively, but did not affect the actions of 3,5-dihydroxyphenylglycine (DHPG), an agonist for group-I mGluRs. These three groups of mGluRs are all endogenously activated during the inspiratory phase. We conclude that three groups of mGluRs are functionally expressed in the phrenic nucleus and that their activation modulates PMN excitability via distinct mechanisms, with group-I acting at postsynaptic sites and group-II and group-III acting at presynaptic sites. PMID- 10377330 TI - N-type calcium channels and their regulation by GABAB receptors in axons of neonatal rat optic nerve. AB - Axons of neonatal rat optic nerves exhibit fast calcium transients in response to brief action potential stimulation. In response to one to four closely spaced action potentials, evoked calcium transients showed a fast-rising phase followed by a decay with a time constant of approximately 2-3 sec. By selective staining of axons or glial cells with calcium dyes, it was shown that the evoked calcium transient originated from axons. The calcium transient was caused by influx because it was eliminated when bath calcium was removed. Pharmacological profile studies with calcium channel subtype-specific peptides suggested that 58% of the evoked calcium influx was accounted for by N-type calcium channels, whereas L- and P/Q-type calcium channels had little, if any, contribution. The identity of the residual calcium influx remains unclear. GABA application caused a dramatic reduction of the amplitude of the action potential and the associated calcium influx. When GABAA receptors were blocked by bicuculline, the inhibitory effect of GABA on the action potential was eliminated, whereas that on the calcium influx was not, indicating involvement of GABAB receptors. Indeed, the calcium influx was inhibited by the GABAB receptor agonist baclofen. This baclofen effect was occluded by a previous block of N-type calcium channels and was unaffected by the broad-spectrum K+ channel blocker 4-AP. We conclude that neonatal rat optic nerve axons express N-type calcium channels, which are subjected to regulation by G-protein-coupled GABAB receptors. We suggest that receptor-mediated inhibition of axonal calcium channels plays a protective role in neonatal anoxic and/or ischemic injury. PMID- 10377331 TI - A zinc-dependent Cl- current in neuronal somata. AB - Extracellular Zn2+ modulates current passage through voltage- and neurotransmitter-gated ion channels, at concentrations less than, or near, those produced by release at certain synapses. Electrophysiological effects of cytoplasmic Zn2+ are less well understood, and effects have been observed at concentrations that are orders of magnitude greater than those found in resting and stimulated neurons. To examine whether and how neurons are affected by lower levels of cytoplasmic Zn2+, we tested the effect of Zn2+-selective chelators, Zn2+-preferring ionophores, and exogenous Zn2+ on neuronal somata during whole cell patch-clamp recordings. We report here that cytoplasmic zinc facilitates the downward regulation of a background Cl- conductance by an endogenous protein kinase C (PKC) in fish retinal ganglion cell somata and that this regulation is maintained if nanomolar levels of free Zn2+ are available. This regulation has not been described previously in any tissue, as other Cl- currents have been described as reduced by PKC alone, reduced by Zn2+ alone, or reduced by both independently. Moreover, control of cation currents by a zinc-dependent PKC has not been reported previously. The regulation we have observed thus provides the first electrophysiological measurements consistent with biochemical measurements of zinc-dependent PKC activity in other systems. These results suggest that contributions of background Cl- conductances to electrical properties of neurons are susceptible to modulation. PMID- 10377332 TI - Calcium-activated potassium conductances contribute to action potential repolarization at the soma but not the dendrites of hippocampal CA1 pyramidal neurons. AB - Evidence is accumulating that voltage-gated channels are distributed nonuniformly throughout neurons and that this nonuniformity underlies regional differences in excitability within the single neuron. Previous reports have shown that Ca2+, Na+, A-type K+, and hyperpolarization-activated, mixed cation conductances have varying distributions in hippocampal CA1 pyramidal neurons, with significantly different densities in the apical dendrites compared with the soma. Another important channel mediates the large-conductance Ca2+-activated K+ current (IC), which is responsible in part for repolarization of the action potential (AP) and generation of the afterhyperpolarization that follows the AP recorded at the soma. We have investigated whether this current is activated by APs retrogradely propagating in the dendrites of hippocampal pyramidal neurons using whole-cell dendritic patch-clamp recording techniques. We found no IC activation by back propagating APs in distal dendritic recordings. Dendritic APs activated IC only in the proximal dendrites, and this activation decayed within the first 100-150 micrometer of distance from the soma. The decay of IC in the proximal dendrites occurred despite AP amplitude, plus presumably AP-induced Ca2+ influx, that was comparable with that at the soma. Thus we conclude that IC activation by action potentials is nonuniform in the hippocampal pyramidal neuron, which may represent a further example of regional differences in neuronal excitability that are determined by the nonuniform distribution of voltage-gated channels in dendrites. PMID- 10377333 TI - kappa- and mu-opioids reverse the somatostatin inhibition of Ca2+ currents in ciliary and dorsal root ganglion neurons. AB - Neuromodulators, including transmitters and peptides, modify neuronal excitability. In most neurons, multiple neuromodulator receptors are present on a single cell. Previous work has demonstrated either occlusive or additive effects when two neuromodulators that target the same ion channel are applied together. In this study, we characterize the modulation of Ca2+ and K+ channels in embryonic chick ciliary ganglion neurons by somatostatin (Som) and opioids, including the effects of these neuromodulators when applied in combination. We report a modulation of calcium current by kappa- or mu-opioids that can prevent Som effects when applied before Som and can replace Som effects when applied after Som. We term these effects demodulation because they do not have the characteristics of simple occlusion but rather represent a dominant effect of opioid-mediated modulation of calcium channels over Som-mediated modulation. These opioid effects persist in the presence of kinase and phosphatase inhibitors, as well as after alteration of the intracellular Ca2+ concentration. Furthermore, they are present in both whole-cell and perforated-patch recording configurations. These effects of opioids on Som-mediated modulation do not seem to be mediated by a general uncoupling of Som receptors from G-protein-coupled signaling systems because K+ current modulation by Som can persist in the presence of opioids. Demodulation by opioids was also observed in dorsal root ganglion neurons on the modulation of calcium current by GABA and norepinephrine (NE). In both preparations, this demodulatory interaction occurred between voltage-independent (opioids) and voltage-dependent (Som, GABA, and NE) modulatory pathways. PMID- 10377334 TI - Selective excitation of subtypes of neocortical interneurons by nicotinic receptors. AB - The cellular mechanisms by which neuronal nicotinic cholinergic receptors influence many aspects of physiology and pathology in the neocortex remain primarily unknown. Whole-cell recordings and single-cell reverse transcription (RT)-PCR were combined to analyze the effect of nicotinic receptor agonists on different types of neurons in acute slices of rat neocortex. Nicotinic receptor agonists had no effect on pyramidal neurons and on most types of interneurons, including parvalbumin-expressing fast spiking interneurons and somatostatin expressing interneurons, but selectively excited a subpopulation of interneurons coexpressing the neuropeptides vasoactive intestinal peptide (VIP) and cholecystokinin. This excitation persisted in the presence of glutamate, GABA, and muscarinic receptor antagonists and in the presence of tetrodotoxin and low extracellular calcium, suggesting that the depolarization was mediated through the direct activation of postsynaptic nicotinic receptors. The responses were blocked by the nicotinic receptor antagonists dihydro-beta-erythroidine and mecamylamine and persisted in the presence of the alpha7 selective nicotinic receptor antagonist methyllycaconitine, suggesting that the involved nicotinic receptors lacked the alpha7 subunit. Single-cell RT-PCR analysis indicated that the majority of the interneurons that responded to nicotinic stimulation coexpressed the alpha4, alpha5, and beta2 nicotinic receptor subunits. Therefore, these results provide a role for non-alpha7 nicotinic receptors in the selective excitation of a subpopulation of neocortical interneurons. Because the neocortical interneurons expressing VIP have been proposed previously to regulate regional cortical blood flow and metabolism, these results also provide a cellular basis for the neuronal regulation of cortical blood flow mediated by acetylcholine. PMID- 10377335 TI - Interleukin-6 (IL-6) production by astrocytes: autocrine regulation by IL-6 and the soluble IL-6 receptor. AB - In the CNS, astrocytes are a major inducible source of interleukin-6 (IL-6). Although IL-6 has beneficial effects in the CNS because of its neurotrophic properties, its overexpression is generally detrimental, adding to the pathophysiology associated with CNS disorders. Many factors have been shown to induce IL-6 expression by astrocytes, particularly the cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1beta). However, the role of IL-6 in its own regulation in astrocytes has not been determined. In this study, we examined the influence of IL-6 alone or in combination with TNF-alpha or IL 1beta on IL-6 expression. IL-6 alone had no effect on IL-6 expression; however, the addition of the soluble IL-6 receptor (sIL-6R) induced IL-6 transcripts. Addition of TNF-alpha or IL-1beta plus IL-6/sIL-6R led to synergistic increases in IL-6 expression. This synergy also occurred in the absence of exogenously added IL-6, attributable to TNF-alpha- or IL-1beta-induced endogenous IL-6 protein production. IL-6 upregulation seen in the presence of TNF-alpha or IL 1beta plus IL-6/sIL-6R was transcriptional, based on nuclear run-on analysis. Experiments were extended to other IL-6 family members to determine their role in IL-6 regulation in astrocytes. Oncostatin M (OSM) induced IL-6 alone and synergized with TNF-alpha for enhanced expression. These results demonstrate that IL-6/sIL-6R and OSM play an important role in the regulation of IL-6 expression within the CNS, particularly in conjunction with the proinflammatory cytokines TNF-alpha and IL-1beta. PMID- 10377337 TI - Molecular basis for the inactivation of Ca2+- and voltage-dependent BK channels in adrenal chromaffin cells and rat insulinoma tumor cells. AB - Large-conductance Ca2+- and voltage-dependent potassium (BK) channels exhibit functional diversity not explained by known splice variants of the single Slo alpha-subunit. Here we describe an accessory subunit (beta3) with homology to other beta-subunits of BK channels that confers inactivation when it is coexpressed with Slo. Message encoding the beta3 subunit is found in rat insulinoma tumor (RINm5f) cells and adrenal chromaffin cells, both of which express inactivating BK channels. Channels resulting from coexpression of Slo alpha and beta3 subunits exhibit properties characteristic of native inactivating BK channels. Inactivation involves multiple cytosolic, trypsin-sensitive domains. The time constant of inactivation reaches a limiting value approximately 25-30 msec at Ca2+ of 10 microM and positive activation potentials. Unlike Shaker N terminal inactivation, but like native inactivating BK channels, a cytosolic channel blocker does not compete with the native inactivation process. Finally, the beta3 subunit confers a reduced sensitivity to charybdotoxin, as seen with native inactivating BK channels. Inactivation arises from the N terminal of the beta3 subunit. Removal of the beta3 N terminal (33 amino acids) abolishes inactivation, whereas the addition of the beta3 N terminal onto the beta1 subunit confers inactivation. The beta3 subunit shares with the beta1 subunit an ability to shift the range of voltages over which channels are activated at a given Ca2+. Thus, the beta-subunit family of BK channels regulates a number of critical aspects of BK channel phenotype, including inactivation and apparent Ca2+ sensitivity. PMID- 10377336 TI - Dynamic regulation of expression and phosphorylation of tau by fibroblast growth factor-2 in neural progenitor cells from adult rat hippocampus. AB - The nature of the extracellular signals that regulate the expression and the phosphorylation of the microtubule-associated protein tau, which is aberrantly hyperphosphorylated in Alzheimer disease and other adult-onset neurodegenerative diseases, is not known. We have found that neural progenitor cells from adult rat hippocampus express adult isoforms of tau and that the expression and the phosphorylation of tau are regulated by fibroblast growth factor-2 (FGF-2). Astrocytes that are differentiated from these cells by stimulation with ciliary neurotrophic factor express phosphorylated tau similarly when cultured in the presence of FGF-2. In fetal progenitor cells that express only the fetal tau isoform, expression, but not the phosphorylation, of this protein is regulated by FGF-2 in cultures of higher passages. The FGF-2-mediated tau hyperphosphorylation is inhibited by lithium, an inhibitor of glycogen synthase kinase-3 (GSK-3), but not by inhibitors of mitogen-activated protein kinase or the cyclin-dependent kinases. Furthermore, both GSK-3 activity and the phosphorylation of tau increase when the concentration of FGF-2 is increased up to 40 ng/ml. These results demonstrate that proliferating adult rat hippocampal progenitor cells express adult isoforms of tau stably and that FGF-2 upregulates the expression and, by upregulating GSK-3 activity, the phosphorylation of tau. PMID- 10377338 TI - The concentration of synaptically released glutamate outside of the climbing fiber-Purkinje cell synaptic cleft. AB - AMPA receptors and glutamate transporters expressed by cerebellar Bergmann glial cells are activated by neurotransmitter released from climbing fibers (). Based on anatomical evidence, this is most likely the result of glutamate diffusing out of the climbing fiber-Purkinje cell synaptic clefts (). We used the change in the EC50 of the Bergmann glia AMPA receptors produced by cyclothiazide (CTZ) to estimate the concentration of glutamate reached at the glial membrane. The decrease of the EC50 gives rise to a concentration-dependent potentiation of the AMPA receptor-mediated responses (). By comparing the increase in amplitude of the AMPA receptor response in the Bergmann glia (840 +/- 240%; n = 8) with the shift in the glutamate dose-response curve measured in excised patches (EC50, 1810 microM in control vs 304 microM in CTZ), we estimate that the extrasynaptic transmitter concentration reaches 160-190 microM. This contrasts with the concentration in the synaptic cleft, thought to rapidly rise above 1 mM, but is still high enough to activate glutamate receptors. These results indicate that the sphere of influence of synaptically released glutamate can extend beyond the synaptic cleft. PMID- 10377339 TI - The electrical properties of auditory hair cells in the frog amphibian papilla. AB - The amphibian papilla (AP) is the principal auditory organ of the frog. Anatomical and neurophysiological evidence suggests that this hearing organ utilizes both mechanical and electrical (hair cell-based) frequency tuning mechanisms, yet relatively little is known about the electrophysiology of AP hair cells. Using the whole-cell patch-clamp technique, we have investigated the electrical properties and ionic currents of isolated hair cells along the rostrocaudal axis of the AP. Electrical resonances were observed in the voltage response of hair cells harvested from the rostral and medial, but not caudal, regions of the AP. Two ionic currents, ICa and IK(Ca), were observed in every hair cell; however, their amplitudes varied substantially along the epithelium. Only rostral hair cells exhibited an inactivating potassium current (IA), whereas an inwardly rectifying potassium current (IK1) was identified only in caudal AP hair cells. Electrically tuned hair cells exhibited resonant frequencies from 50 to 375 Hz, which correlated well with hair cell position and the tonotopic organization of the papilla. Variations in the kinetics of the outward current contribute substantially to the determination of resonant frequency. ICa and IK(Ca) amplitudes increased with resonant frequency, reducing the membrane time constant with increasing resonant frequency. We conclude that a tonotopically organized hair cell substrate exists to support electrical tuning in the rostromedial region of the frog amphibian papilla and that the cellular mechanisms for frequency determination are very similar to those reported for another electrically tuned auditory organ, the turtle basilar papilla. PMID- 10377340 TI - Involvement of cGMP-dependent protein kinase in adrenergic potentiation of transmitter release from the calyx-type presynaptic terminal. AB - I have previously reported that norepinephrine (NE) induces a sustained potentiation of transmitter release in the chick ciliary ganglion through a mechanism pharmacologically distinct from any known adrenergic receptors. Here I report that the adrenergic potentiation of transmitter release was enhanced by a phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine (IBMX) and by zaprinast, an inhibitor of cGMP-selective phosphodiesterase. Exogenous application of the membrane-permeable cGMP, 8-bromo-cGMP (8Br-cGMP), potentiated the quantal transmitter release, and after potentiation, the addition of NE was no longer effective. On the other hand, 8Br-cAMP neither potentiated the transmitter release nor occluded the NE-induced potentiation. The NE-induced potentiation was blocked by neither nitric oxide (NO) synthase inhibitor nor NO scavenger. The quantal transmitter release was not potentiated by NO donors, e.g., sodium nitroprusside. The NE-induced potentiation and its enhancement by IBMX was antagonized by two inhibitors of protein kinase G (PKG), Rp isomer of 8-(4 chlorophenylthio) guanosine-3', 5'-cyclic monophosphorothioate and KT5823. As with NE-induced potentiation, the effects of 8Br-cGMP on both the resting intraterminal [Ca2+] ([Ca2+]i) and the action potential-dependent increment of [Ca2+]i (DeltaCa) in the presynaptic terminal were negligible. The reduction of the paired pulse ratio of EPSC is consistent with the notion that the NE- and cGMP-dependent potentiation of transmitter release was attributable mainly to an increase of the exocytotic fusion probability. These results indicate that NE binds to a novel adrenergic receptor that activates guanylyl cyclase and that accumulation of cGMP activates PKG, which may phosphorylate a target protein involved in the exocytosis of synaptic vesicles. PMID- 10377342 TI - Fast excitatory synaptic transmission mediated by nicotinic acetylcholine receptors in Drosophila neurons. AB - Difficulty in recording from single neurons in vivo has precluded functional analyses of transmission at central synapses in Drosophila, where the neurotransmitters and receptors mediating fast synaptic transmission have yet to be identified. Here we demonstrate that spontaneously active synaptic connections form between cultured neurons prepared from wild-type embryos and provide the first direct evidence that both acetylcholine and GABA mediate fast interneuronal synaptic transmission in Drosophila. The predominant type of fast excitatory transmission between cultured neurons is mediated by nicotinic acetylcholine receptors (nAChRs). Detailed analysis of cholinergic transmission reveals that spontaneous EPSCs (sEPSCs) are composed of both evoked and action potential independent [miniature EPSC (mEPSC)] components. The mEPSCs are characterized by a broad, positively skewed amplitude histogram in which the variance is likely to reflect differences in the currents induced by single quanta. Biophysical characteristics of the cholinergic mEPSCs include a rapid rise time (0.6 msec) and decay (tau = 2 msec). Regulation of mEPSC frequency by external calcium and cobalt suggests that calcium influx through voltage-gated channels influences the probability of ACh release. In addition, brief depolarization of the cultures with KCl can induce a calcium-dependent increase in sEPSC frequency that persists for up to 3 hr after termination of the stimulus, illustrating one form of plasticity at these cholinergic synapses. These data demonstrate that cultured embryonic neurons, amenable to both genetic and biochemical manipulations, present a unique opportunity to define genes/signal transduction cascades involved in functional regulation of fast excitatory transmission at interneuronal cholinergic synapses in Drosophila. PMID- 10377341 TI - Voltage-dependent neuromodulation of Na+ channels by D1-like dopamine receptors in rat hippocampal neurons. AB - Activation of D1-like dopamine (DA) receptors reduces peak Na+ current in acutely isolated hippocampal neurons through phosphorylation of the alpha subunit of the Na+ channel by cAMP-dependent protein kinase (PKA). Here we report that neuromodulation of Na+ currents by DA receptors via PKA is voltage-dependent in the range of -110 to -70 mV and is also sensitive to concurrent activation of protein kinase C (PKC). Depolarization enhanced the ability of D1-like DA receptors to reduce peak Na+ currents via the PKA pathway. Similar voltage dependent modulation was observed when PKA was activated directly with the membrane-permeant PKA activator DCl-cBIMPS (cBIMPS; 20 microM), indicating that the membrane potential dependence occurs downstream of PKA. PKA activation caused only a small (-2.9 mV) shift in the voltage dependence of steady-state inactivation and had no effect on slow inactivation or on the rates of entry into the fast or slow inactivated states, suggesting that another mechanism is responsible for coupling of membrane potential changes to PKA modulation. Activation of PKC with a low concentration of the membrane-permeant diacylglycerol analog oleylacetyl glycerol also potentiated modulation by SKF 81297 or cBIMPS, and these effects were most striking at hyperpolarized membrane potentials where PKA modulation was not stimulated by membrane depolarization. Thus, activation of D1-like DA receptors causes a strong reduction in Na+ current via the PKA pathway, but it is effective primarily when it is combined with depolarization or activation of PKC. The convergence of these three distinct signaling modalities on the Na+ channel provides an intriguing mechanism for integration of information from multiple signaling pathways in the hippocampus and CNS. PMID- 10377343 TI - Alternative splicing of a short cassette exon in alpha1B generates functionally distinct N-type calcium channels in central and peripheral neurons. AB - The N-type Ca channel alpha1B subunit is localized to synapses throughout the nervous system and couples excitation to release of neurotransmitters. In a previous study, two functionally distinct variants of the alpha1B subunit were identified, rnalpha1B-b and rnalpha1B-d, that differ at two loci;four amino acids [SerPheMetGly (SFMG)] in IIIS3-S4 and two amino acids [GluThr (ET)] in IVS3-S4. These variants are reciprocally expressed in rat brain and sympathetic ganglia (). We now show that the slower activation kinetics of rnalpha1B-b (DeltaSFMG/+ET) compared with rnalpha1B-d (+SFMG/DeltaET) channels are fully accounted for by the insertion of ET in IVS3-S4 and not by the lack of SFMG in IIIS3-S4. We also show that the inactivation kinetics of these two variants are indistinguishable. Through genomic analysis we identify a six-base cassette exon that encodes the ET site and with ribonuclease protection assays demonstrate that the expression of this mini-exon is essentially restricted to alpha1B RNAs of peripheral neurons. We also show evidence for regulated alternative splicing of a six-base exon encoding NP in the IVS3-S4 linker of the closely related alpha1A gene and establish that residues NP can functionally substitute for ET in domain IVS3-S4 of alpha1B. The selective expression of functionally distinct Ca channel splice variants of alpha1B and alpha1A subunits in different regions of the nervous system adds a new dimension of diversity to voltage-dependent Ca signaling in neurons that may be important for optimizing action potential dependent transmitter release at different synapses. PMID- 10377344 TI - The native rat olfactory cyclic nucleotide-gated channel is composed of three distinct subunits. AB - Cyclic nucleotide-gated (CNG) channels play central roles in visual and olfactory signal transduction. In the retina, rod photoreceptors express the subunits CNCalpha1 and CNCbeta1a. In cone photoreceptors, only CNCalpha2 expression has been demonstrated so far. Rat olfactory sensory neurons (OSNs) express two homologous subunits, here designated CNCalpha3 and CNCalpha4. This paper describes the characterization of CNCbeta1b, a third subunit expressed in OSNs and establishes it as a component of the native channel. CNCbeta1b is an alternate splice form of the rod photoreceptor CNCbeta1a subunit. Analysis of mRNA and protein expression together suggest co-expression of all three subunits in sensory cilia of OSNs. From single-channel analyses of native rat olfactory channels and of channels expressed heterologously from all possible combinations of the CNCalpha3, -alpha4, and -beta1b subunits, we conclude that the native CNG channel in OSNs is composed of all three subunits. Thus, CNG channels in both rod photoreceptors and olfactory sensory neurons result from coassembly of specific alpha subunits with various forms of an alternatively spliced beta subunit. PMID- 10377345 TI - The Caenorhabditis elegans unc-49 locus encodes multiple subunits of a heteromultimeric GABA receptor. AB - Ionotropic GABA receptors generally require the products of three subunit genes. By contrast, the GABA receptor needed for locomotion in Caenorhabditis elegans requires only the unc-49 gene. We cloned unc-49 and demonstrated that it possesses an unusual overlapping gene structure. unc-49 contains a single copy of a GABA receptor N terminus, followed by three tandem copies of a GABA receptor C terminus. Using a single promoter, unc-49 generates three distinct GABAA receptor like subunits by splicing the N terminus to each of the three C-terminal repeats. This organization suggests that the three UNC-49 subunits (UNC-49A, UNC-49B, and UNC-49C) are coordinately rescued and therefore might coassemble to form a heteromultimeric GABA receptor. Surprisingly, only UNC-49B and UNC-49C are expressed at high levels, whereas UNC-49A expression is barely detectable. Green fluorescent protein-tagged UNC-49B and UNC-49C subunits are coexpressed in muscle cells and are colocalized to synaptic regions. UNC-49B and UNC-49C also coassemble efficiently in Xenopus oocytes and HEK-293 cells to form a heteromeric GABA receptor. Together these data argue that UNC-49B and UNC-49C coassemble at the C. elegans neuromuscular junction. Thus, C. elegans is able to encode a heteromeric GABA receptor with a single locus. PMID- 10377346 TI - Mutant presenilin-1 induces apoptosis and downregulates Akt/PKB. AB - Most early onset cases of familial Alzheimer's disease (AD) are caused by mutations in presenilin-1 (PS1) and presenilin-2 (PS2). These mutations lead to increased beta-amyloid formation and may induce apoptosis in some model systems. Using primary cultured hippocampal neurons (HNs) and rat pheochromocytoma (PC12) cells transiently transfected with replication-defective recombinant adenoviral vectors expressing wild-type or mutant PS1, we demonstrate that mutant PS1s induce apoptosis, downregulate the survival factor Akt/PKB, and affect several Akt/PKB downstream targets, including glycogen synthase kinase-3beta and beta catenin. Expression of a constitutively active Akt/PKB rescues HNs from mutant PS1-induced neuronal cell death, suggesting a potential therapeutic target for AD. Downregulation of Akt/PKB may be a mechanism by which mutant PS1 induces apoptosis and may play a role in the pathogenesis of familial AD. PMID- 10377347 TI - Selective disruption of "late onset" sagittal banding patterns by ectopic expression of engrailed-2 in cerebellar Purkinje cells. AB - To explore the role of Engrailed proteins in development of the cerebellum, Engrailed-2 (En-2) was ectopically expressed in cerebellar Purkinje cells from the late embryonic stage into adulthood. The fundamental organization of Purkinje cell sagittal zones as revealed by the "early onset" markers L7-beta-gal and cadherin-8 was found to be virtually identical to that in wild type. In contrast, "late onset" sagittal banding patterns revealed by Purkinje cell markers zebrin I, zebrin II, and 9-O-acetyl GD3 Ganglioside (P-Path), and the granule cell marker NADPH-diaphorase, were disrupted. In general, although some evidence of banding was still detectable, boundaries defined by the latter markers were poorly defined, and the patterns overall took on a diffuse appearance. In parallel with the changes in late onset markers, anterograde tracing of spinocerebellar axons revealed a general diffusion of the mossy fiber projection pattern in lobule VIII and the anterior lobe. These observations suggest that at least two separate mediolateral boundary systems exist in the cerebellum, and these are differentially affected by ectopic En-2 expression. Alternatively, one boundary system exists that remains primarily intact in the mutant, but recognition of this system by a set of late developmental events is perturbed. PMID- 10377348 TI - Voltage-activated K+ channels and membrane depolarization regulate accumulation of the cyclin-dependent kinase inhibitors p27(Kip1) and p21(CIP1) in glial progenitor cells. AB - Neural cell development is regulated by membrane ion channel activity. We have previously demonstrated that cell membrane depolarization with veratridine or blockage of K+ channels with tetraethylammonium (TEA) inhibit oligodendrocyte progenitor (OP) proliferation and differentiation (); however the molecular events involved are largely unknown. Here we show that forskolin (FSK) and its derivative dideoxyforskolin (DFSK) block K+ channels in OPs and inhibit cell proliferation. The antiproliferative effects of TEA, FSK, DFSK, and veratridine were attributable to OP cell cycle arrest in G1 phase. In fact, (1) cyclin D accumulation in synchronized OP cells was not affected by K+ channel blockers or veratridine; (2) these agents prevented OP cell proliferation only if present during G1 phase; and (3) G1 blockers, such as rapamycin and deferoxamine, mimicked the anti-proliferative effects of K+ channel blockers. DFSK also prevented OP differentiation, whereas FSK had no effect. Blockage of K+ channels and membrane depolarization also caused accumulation of the cyclin-dependent kinase inhibitors p27(Kip1) and p21(CIP1) in OP cells. The antiproliferative effects of K+ channel blockers and veratridine were still present in OP cells isolated from INK4a-/- mice, lacking the cyclin-dependent kinase inhibitors p16(INK4a) and p19(ARF). Our results demonstrate that blockage of K+ channels and cell depolarization induce G1 arrest in the OP cell cycle through a mechanism that may involve p27(Kip1) and p21(CIP1) and further support the conclusion that OP cell cycle arrest and differentiation are two uncoupled events. PMID- 10377349 TI - Functionally antagonistic interactions between the TrkA and p75 neurotrophin receptors regulate sympathetic neuron growth and target innervation. AB - In this report, we provide evidence that NGF and BDNF have functionally antagonistic actions on sympathetic neuron growth and target innervation, with NGF acting via TrkA to promote growth and BDNF via p75NTR to inhibit growth. Specifically, in cultured sympathetic neurons that themselves synthesize BDNF, exogenous BDNF inhibits and function-blocking BDNF antibodies enhance process outgrowth. Both exogenous and autocrine BDNF mediate this effect via p75NTR because (1) BDNF does not inhibit growth of neurons lacking p75NTR, (2) function blocking p75NTR antibodies enhance NGF-mediated growth, and (3) p75NTR-/- sympathetic neurons grow more robustly in response to NGF than do their wild-type counterparts. To determine the physiological relevance of this functional antagonism, we examined the pineal gland, a well defined sympathetic target organ. BDNF is present in the pineal gland during target innervation, and incoming sympathetic axons are p75NTR positive. Moreover, the pineal glands of BDNF+/- and BDNF-/- mice are hyperinnervated with sympathetic fibers, and tyrosine hydroxylase (TH) levels are elevated. Increased tyrosine hydroxylase is also observed in the BDNF+/- carotid artery, another sympathetic neuron target. Thus, BDNF, made by sympathetic neurons and/or their target organs, acts via p75NTR to antagonize NGF-mediated growth and target innervation, suggesting that sympathetic target innervation is determined by the balance of positively and negatively acting neurotrophins present in developing and potentially mature targets. PMID- 10377350 TI - Expression of the striatal DARPP-32/ARPP-21 phenotype in GABAergic neurons requires neurotrophins in vivo and in vitro. AB - The medium spiny neuron (MSN) is the major output neuron of the caudate nucleus and uses GABA as its primary neurotransmitter. A majority of MSNs coexpress DARPP 32 and ARPP-21, two dopamine and cyclic AMP-regulated phosphoproteins, and most of the matrix neurons express calbindin. DARPP-32 is the most commonly used MSN marker, but previous attempts to express this gene in vitro have failed. In this study we found that DARPP-32 is expressed in <12% of E13- or E17-derived striatal neurons when they are grown in defined media at high or low density in serum, dopamine, or Neurobasal/N2 (Life Technologies), and ARPP-21 is expressed in <1%. The percentage increases to 25% for DARPP-32 and 10% for ARPP-21 when the same cells are grown in Neurobasal/B27 (Life Technologies) for 7 d. After growth in Neurobasal/B27 plus brain-derived neurotrophic factor (BDNF) for 7 d, E13-derived MSNs are 53.7% DARPP-32-positive and 29. 0% ARPP-21-positive; E17-derived MSNs are 66.8% DARPP-32-positive and 51.5% ARPP-21-positive. The percentage of calbindin-positive neurons also is increased under these conditions. Finally, ARPP-21 expression is reduced in mice with a targeted deletion of the BDNF gene. We conclude that BDNF is required for the maturation of a large subset of patch and matrix MSNs in vivo and in vitro. In addition, we introduce a culture system in which highly differentiated MSNs may be generated, maintained, and studied. PMID- 10377351 TI - Immortalized human dorsal root ganglion cells differentiate into neurons with nociceptive properties. AB - A renewable source of human sensory neurons would greatly facilitate basic research and drug development. We had established previously conditionally immortalized human CNS cell lines that can differentiate into functional neurons (). We report here the development of an immortalized human dorsal root ganglion (DRG) clonal cell line, HD10.6, with a tetracycline-regulatable v-myc oncogene. In the proliferative condition, HD10.6 cells have a doubling time of 1.2 d and exhibit a neuronal precursor morphology. After differentiation of clone HD10.6 for 7 d in the presence of tetracycline, v-myc expression was suppressed, and >50% of the cells exhibited typical neuronal morphology, stained positively for neuronal cytoskeletal markers, and fired action potentials in response to current injection. Furthermore, this cell line was fate-restricted to a neuronal phenotype; even in culture conditions that promote Schwann cell or smooth muscle differentiation of neural crest stem cells, HD10.6 differentiated exclusively into neurons. Moreover, differentiated HD10.6 cells expressed sensory neuron associated transcription factors and exhibited capsaicin sensitivity. Taken together, these data indicate that we have established an immortalized human DRG cell line that can differentiate into sensory neurons with nociceptive properties. The cell line HD10.6 represents the first example of a human sensory neuronal line and will be valuable for basic research, as well as for the discovery of novel drug targets and clinical candidates. PMID- 10377352 TI - Developmental requirement of gp130 signaling in neuronal survival and astrocyte differentiation. AB - gp130 is a signal-transducing receptor component used in common by the interleukin-6 (IL-6) family of hematopoietic and neurotrophic cytokines, including IL-6, IL-11, leukemia-inhibitory factor, ciliary neurotrophic factor, oncostatin-M, and cardiotrophin-1. We have examined in this study a role of gp130 in the nervous system by analyzing developmental cell death of several neuronal populations and the differentiation of astrocytes in gp130-deficient mice. A significant reduction was observed in the number of sensory neurons in L5 dorsal root ganglia and motoneurons in the facial nucleus, the nucleus ambiguus, and the lumbar spinal cord in gp130 -/- mice on embryonic day 18.5. On the other hand, no significant neuronal loss was detectable on day 14.5, suggesting a physiological role of gp130 in supporting newly generated neurons during the late phase of development when naturally occurring cell death takes place. Moreover, expression of an astrocyte marker, GFAP, was severely reduced in the brain of gp130 -/- mice. Our data demonstrate that gp130 expression is essential for survival of subgroups of differentiated motor and sensory neurons and for the differentiation of major populations of astrocytes in vivo. PMID- 10377353 TI - Increased synchronization of neuromagnetic responses during conscious perception. AB - In binocular rivalry, the observer views two incongruent images, one through each eye, but is conscious of only one image at a time. The image that is perceptually dominant alternates every few seconds. We used this phenomenon to investigate neural correlates of conscious perception. We presented a red vertical grating to one eye and a blue horizontal grating to the other eye, with each grating continuously flickering at a distinct frequency (the frequency tag for that stimulus). Steady-state magnetic fields were recorded with a 148 sensor whole head magnetometer while the subjects reported which grating was perceived. The power of the steady-state magnetic field at the frequency associated with a grating typically increased at multiple sensors when the grating was perceived. Changes in power related to perceptual dominance, presumably reflecting local neural synchronization, reached statistical significance at several sensors, including some positioned over occipital, temporal, and frontal cortices. To identify changes in synchronization between distinct brain areas that were related to perceptual dominance, we analyzed coherence between pairs of widely separated sensors. The results showed that when the stimulus was perceived there was a marked increase in both interhemispheric and intrahemispheric coherence at the stimulus frequency. This study demonstrates a direct correlation between the conscious perception of a visual stimulus and the synchronous activity of large populations of neocortical neurons as reflected by stimulus-evoked steady-state neuromagnetic fields. PMID- 10377354 TI - Different proctolin neurons elicit distinct motor patterns from a multifunctional neuronal network. AB - Distinct motor patterns are selected from a multifunctional neuronal network by activation of different modulatory projection neurons. Subsets of these projection neurons can contain the same neuromodulator(s), yet little is known about the relative influence of such neurons on network activity. We have addressed this issue in the stomatogastric nervous system of the crab Cancer borealis. Within this system, there is a neuronal network in the stomatogastric ganglion (STG) that produces many versions of the pyloric and gastric mill rhythms. These different rhythms result from activation of different projection neurons that innervate the STG from neighboring ganglia and modulate STG network activity. Three pairs of these projection neurons contain the neuropeptide proctolin. These include the previously identified modulatory proctolin neuron and modulatory commissural neuron 1 (MCN1) and the newly identified modulatory commissural neuron 7 (MCN7). We document here that each of these neurons contains a unique complement of cotransmitters and that each of these neurons elicits a distinct version of the pyloric motor pattern. Moreover, only one of them (MCN1) also elicits a gastric mill rhythm. The MCN7-elicited pyloric rhythm includes a pivotal switch by one STG network neuron from playing a minor to a major role in motor pattern generation. Therefore, modulatory neurons that share a peptide transmitter can elicit distinct motor patterns from a common target network. PMID- 10377355 TI - Differential regulation of corticotropin-releasing hormone and vasopressin gene transcription in the hypothalamus by norepinephrine. AB - All stress-related inputs are conveyed to the hypothalamus via several brain areas and integrated in the parvocellular division of the paraventricular nucleus (PVN) where corticotropin-releasing hormone (CRH) is synthesized. Arginine vasopressin (AVP) is present in both magnocellular and parvocellular divisions of the PVN, and the latter population of AVP is colocalized with CRH. CRH and AVP are co-secreted in the face of certain stressful stimuli, and synthesis of both peptides is suppressed by glucocorticoid. CRH and AVP stimulate corticotropin (ACTH) secretion synergistically, but the physiological relevance of the dual corticotroph regulation is not understood. Norepinephrine (NE) is a well known neurotransmitter that regulates CRH neurons in the PVN. We explored the mode of action of NE on CRH and AVP gene transcription in the PVN to examine the effect of the neurotransmitter on multiple genes that are responsible for a common physiological function. After NE injection into the PVN of conscious rats, CRH heteronuclear (hn) RNA increased rapidly and markedly in the parvocellular division of the PVN. AVP hnRNA did not change significantly in either the parvocellular or magnocellular division of the PVN after NE injection. The present results show that the transcription of CRH and AVP genes is differentially regulated by NE, indicating the complexity of neurotransmitter regulation of multiple releasing hormone genes in a discrete hypothalamic neuronal population. PMID- 10377356 TI - Different contributions of the human amygdala and ventromedial prefrontal cortex to decision-making. AB - The somatic marker hypothesis proposes that decision-making is a process that depends on emotion. Studies have shown that damage of the ventromedial prefrontal (VMF) cortex precludes the ability to use somatic (emotional) signals that are necessary for guiding decisions in the advantageous direction. However, given the role of the amygdala in emotional processing, we asked whether amygdala damage also would interfere with decision-making. Furthermore, we asked whether there might be a difference between the roles that the amygdala and VMF cortex play in decision-making. To address these two questions, we studied a group of patients with bilateral amygdala, but not VMF, damage and a group of patients with bilateral VMF, but not amygdala, damage. We used the "gambling task" to measure decision-making performance and electrodermal activity (skin conductance responses, SCR) as an index of somatic state activation. All patients, those with amygdala damage as well as those with VMF damage, were (1) impaired on the gambling task and (2) unable to develop anticipatory SCRs while they pondered risky choices. However, VMF patients were able to generate SCRs when they received a reward or a punishment (play money), whereas amygdala patients failed to do so. In a Pavlovian conditioning experiment the VMF patients acquired a conditioned SCR to visual stimuli paired with an aversive loud sound, whereas amygdala patients failed to do so. The results suggest that amygdala damage is associated with impairment in decision-making and that the roles played by the amygdala and VMF in decision-making are different. PMID- 10377357 TI - Chronic pain is associated with increased TrkA immunoreactivity in spinoreticular neurons. AB - Repetitive noxious stimulation leads to permanent adaptive changes of central pathways involved in the genesis and integration of nociception. Several classes of neurotrophic factors that affect brain plasticity are also involved in the regulation of sensory functions in adulthood. To investigate a putative role of nerve growth factor (NGF) in central plasticity linked to chronic pain, modifications in immunoreactivity (IR) for the high-affinity NGF receptor, TrkA, were studied at spinal levels in a rat model of inflammatory chronic pain, adjuvant-induced arthritis (AIA). We report a specific increase in the number of TrkA-IR profiles in laminae V-VI at lumbar levels L3 and L4 in arthritic rats. Tract tracing using FluoroGold injections in the ventrobasal complex of the thalamus and in the brainstem showed that these increased TrkA-IR profiles are spinoreticular neurons. Dual labeling with calcitonin gene-related peptide or substance P showed that TrkA-IR neurons were mainly located in projection fields of small- to medium-sized primary afferent fibers, which convey nociceptive inputs. These results suggest that TrkA-containing neurons of the spinal dorsal horn participate in the first central relay of transmission of nociceptive information to supraspinal centers. Enhanced numbers of TrkA-IR neurons during AIA strongly support the hypothesis of a participation of NGF in adaptive mechanisms of central nociceptive pathways observed in chronic pain states. PMID- 10377358 TI - Prospective coding for objects in primate prefrontal cortex. AB - We examined neural activity in prefrontal (PF) cortex of monkeys performing a delayed paired associate task. Monkeys were cued with a sample object. Then, after a delay, a test object was presented. If the test object was the object associated with the sample during training (i.e., its target), they had to release a lever. Monkeys could bridge the delay by remembering the sample (a sensory-related code) and/or thinking ahead to the expected target (a prospective code). Examination of the monkeys' behavior suggested that they were relying on a prospective code. During and shortly after sample presentation, neural activity in the lateral PF cortex primarily reflected the sample. Toward the end of the delay, however, PF activity began to reflect the anticipated target, which indicated a prospective code. These results provide further confirmation that PF cortex does not simply buffer incoming visual inputs, but instead selectively processes information relevant to current behavioral demands, even when this information must be recalled from long-term memory. PMID- 10377359 TI - Brain mechanisms of propofol-induced loss of consciousness in humans: a positron emission tomographic study. AB - In the present study, we used positron emission tomography to investigate changes in regional cerebral blood flow (rCBF) during a general anesthetic infusion set to produce a gradual transition from the awake state to unconsciousness. Five right-handed human volunteers participated in the study. They were given propofol with a computer-controlled infusion pump to achieve three stable levels of plasma concentrations corresponding to mild sedation, deep sedation, and unconsciousness, the latter defined as unresponsiveness to verbal commands. During awake baseline and each of the three levels of sedation, two scans were acquired after injection of an H215O bolus. Global as well as regional CBF were determined and correlated with propofol concentrations. In addition, blood flow changes in the thalamus were correlated with those of the entire scanned volume to determine areas of coordinated changes. In addition to a generalized decrease in global CBF, large regional decreases in CBF occurred bilaterally in the medial thalamus, the cuneus and precuneus, and the posterior cingulate, orbitofrontal, and right angular gyri. Furthermore, a significant covariation between the thalamic and midbrain blood flow changes was observed, suggesting a close functional relationship between the two structures. We suggest that, at the concentrations attained, propofol preferentially decreases rCBF in brain regions previously implicated in the regulation of arousal, performance of associative functions, and autonomic control. Our data support the hypothesis that anesthetics induce behavioral changes via a preferential, concentration-dependent effect on specific neuronal networks rather than through a nonspecific, generalized effect on the brain. PMID- 10377360 TI - Optic flow input to the hippocampal formation from the accessory optic system. AB - Recent studies in rodents have implicated the hippocampal formation in "path integration": the ability to use self-motion cues (ideothesis) to guide spatial behavior. Such models of hippocampal function assume that self-motion information arises from the vestibular system. In the present study we used the retrograde tracer cholera toxin subunit B, the anterograde tracer biotinylated dextran amine, and standard extracellular recording techniques to investigate whether the hippocampal formation [which consists of the hippocampus proper and the area parahippocampalis (Hp/APH) in pigeons] receives information from the accessory optic system (AOS). The AOS is a visual pathway dedicated to the analysis of the "optic flow fields" that result from self-motion. Optic flow constitutes a rich source of ideothetic information that could be used for navigation. Both the nucleus of the basal optic root (nBOR) and nucleus lentiformis mesencephali of the AOS were shown to project to the area ventralis of Tsai (AVT), which in turn was shown to project to the Hp/APH. A smaller direct projection from the nBOR pars dorsalis to the hippocampus was also revealed. During extracellular recording experiments, about half of the cells within the AVT responded to optic flow stimuli. Together these results illustrate that the Hp/APH receives information about self-motion from the AOS. We postulate that this optic flow information is used for path integration. A review of the current literature suggests that an analogous neuronal circuit exists in mammals, but it has simply been overlooked. PMID- 10377361 TI - Arginine analogs modify signal detection by neurons in the visual cortex. AB - Nitric oxide (NO) modulates neurotransmitter release, induction of long-term synaptic potentiation and depression, and activity levels of neurons. However, it is not known whether NO contributes to the ability of the CNS to distinguish sensory signals from background noise and/or extract sensory information with greater reliability. We addressed these questions in the visual cortex, in vivo, using electrophysiological recording and analysis of signal detection from individual neurons. This was combined with microiontophoretic application of arginine analogs that either upregulate or downregulate the brain's endogenous NO generating pathways or compounds that produce exogenous NO. Protocols that enhance NO levels generally increased the number of action potentials per trial evoked by visual stimuli, improved signal detection, and decreased the coefficient of variation of visually evoked responses, whereas NO-reducing protocols predominantly had complementary effects. Control experiments demonstrate that these effects are likely attributable to the specific ability of these arginine compounds to modify NO levels versus other nonspecific effects. Differential effects between neighboring cells and between single-cell receptive subfields suggest that these actions have a significant direct neural component versus exclusively operating indirectly on neurons through the central vascular actions of NO. PMID- 10377362 TI - Differential distribution of intracellular glutamate receptors in dendrites. AB - Glutamate receptors are synthesized in the cell body and transported in intracellular compartments to the target synapse. The objective of the present study was to analyze the intracellular pool of glutamate receptors and determine whether the intracellular pool was related to the synaptic distribution of the receptors. As a model system, we chose the fusiform cell of the dorsal cochlear nucleus for which we have previously demonstrated that receptors are selectively targeted to synapses on apical and basal dendrites. A combination of retrograde tracing and postembedding immunogold labeling was used to quantify intracellular receptors in segments of apical and basal dendrites. Immunolabeling for GluR4 and mGluR1alpha is present at synapses on basal dendrites but not on apical dendrites, whereas immunolabeling for GluR2/3 is present at both populations of synapses. In the analysis of intracellular pools, we find that GluR2/3 is equally distributed in apical and basal dendrites, whereas GluR4 and mGluR1alpha are more concentrated in basal dendrites than in apical dendrites. These findings indicate that the distribution of intracellular receptors is related to that of synaptic receptors and suggest that a mechanism exists in neurons to target proteins to dendritic domains soon after synthesis. We found no evidence for the existence of a pool of intracellular receptors, which could represent a receptor reserve, near the postsynaptic density. Receptors were often found in clusters associated with tubulovesicular membranes of the endoplasmic reticulum, identified with immunoglobulin binding protein (BIP) or calnexin, suggesting that this organelle is involved in receptor transport in dendrites. PMID- 10377363 TI - Diminished viability, growth, and behavioral efficacy of fetal dopamine neuron grafts in aging rats with long-term dopamine depletion: an argument for neurotrophic supplementation. AB - We examined the behavioral and morphological correlates of the response to a single intrastriatal dispersed cell graft of fetal rat ventral mesencephalic tissue in male Fischer-344 rats of varying age (4, 17, and 24-26 months old) and history of mesostriatal dopamine (DA) depletion (1 or 14 months). Our goal was to determine the impact of advancing age and duration of DA depletion in the host on DA graft viability and function. The findings can be summarized as follows. (1) Fetal DA neuron grafts that were effective in completely ameliorating amphetamine induced rotational behavior in young rats with short-term lesions were virtually without effect in aged rats with long-term lesions. Middle-aged rats with long term lesions responded to these grafts with partial behavioral recovery. (2) Age of the host at the time of transplantation, and not duration of DA depletion, was the primary determinant of response to DA grafts. (3) Diminished efficacy of grafts in lesioned aging rats was related to decreased survival and neurite extension of transplanted DA neurons. (4) Co-grafts of DA neurons with Schwann cells as a source of neurotrophic support improved the behavioral outcome of grafts in aged lesioned rats. These findings support the view that the DA depleted striatum of aged rats is an impoverished environment for survival, growth, and function of DA grafts. Consistent with this view, local supplementation of the neurotrophic environment of grafted DA neurons with products of co-grafted Schwann cells, a demonstrated source of neurotrophic activity for embryonic DA neurons, improved graft outcome. PMID- 10377364 TI - Localization of a suprachiasmatic nucleus subregion regulating locomotor rhythmicity. AB - The bilaterally symmetrical suprachiasmatic nuclei (SCN) of the hypothalamus are the loci of the mammalian clock controlling circadian rhythms. Previous studies suggested that all regions of the SCN are equipotential as circadian rhythmicity is sustained after partial ablation, as long as approximately 25% of the nuclei are spared. In contrast to these results, we found that animals bearing partial lesions of the SCN that spared the subregion delimited by cells containing the calcium-binding protein calbindin-D28K (CaBP), sustained circadian locomotor rhythms. Furthermore, there was a correlation between the strength of the rhythm and the number of spared CaBP cells. Partial lesions that destroyed this region but spared other compartments of the SCN resulted in loss of rhythmicity. The next study indicates that transplants of half-SCN grafts that contain CaBP cells restore locomotor rhythms in SCN-lesioned host animals, whereas transplants containing SCN tissue but lacking cells of this subnucleus fail to restore rhythmicity. Finally, there was a correlation between the number of CaBP-positive cells in the graft and the strength of the restored rhythm. Taken together, the results indicate that pacemakers in the region of the CaBP subnucleus are necessary and sufficient for the control of locomotor rhythmicity and that the SCN is functionally heterogeneous. PMID- 10377365 TI - Spontaneous activity of neostriatal cholinergic interneurons in vitro. AB - Neostriatal cholinergic interneurons fire irregularly but tonically in vivo. The summation of relatively few depolarizing potentials and their temporal sequence are thought to underlie spike triggering and the irregularity of action potential timing, respectively. In these experiments we used whole-cell, cell-attached, and extracellular recording techniques to investigate the role of spontaneous synaptic inputs in the generation and patterning of action potentials in cholinergic interneurons in vitro. Cholinergic cells were spontaneously active in vitro at 25 +/- 1 degrees C during whole-cell recording from 2 to 3 week postnatal slices and at 35 +/- 2 degrees C during cell-attached and extracellular recording from 3 to 4 week postnatal slices. A range of firing frequencies and patterns was observed including regular, irregular, and burst firing. Blockade of AMPA and NMDA receptors altered neither the firing rate nor the pattern, and accordingly, voltage-clamp data revealed a very low incidence of spontaneous EPSCs. GABAA receptor antagonists were also ineffective in altering the spiking frequency or pattern owing to minimal inhibitory input in vitro. Functional excitatory and inhibitory inputs to cholinergic cells were disclosed after application of 4-aminopyridine (100 microM), indicating that these synapses are present but not active in vitro. Blockade of D1 or D2 dopamine receptors or muscarinic receptors also failed to influence tonic activity in cholinergic cells. Together these data indicate that cholinergic interneurons are endogenously active and generate action potentials in the absence of any synaptic input. Interspike interval histograms and autocorrelograms generated from unit recordings of cholinergic cells in vitro were indistinguishable from those of tonically active neurons recorded in vivo. Irregular spiking is therefore embedded in the mechanism responsible for endogenous activity. PMID- 10377367 TI - Binocular neurons in V1 of awake monkeys are selective for absolute, not relative, disparity. AB - Most neurophysiological accounts of disparity selectivity in neurons of the primary visual cortex (V1) imply that they are selective for absolute retinal disparities. By contrast, a number of psychophysical observations indicate that relative disparities play a more important role in depth perception. During recordings from disparity selective neurons in area V1 of awake behaving monkeys, we used a disparity feedback loop () to add controlled amounts of absolute disparity to a display containing both absolute and relative disparities. This manipulation changed the absolute disparity of all the visible features in the display but left unchanged the relative disparities signalled by these features. The addition of absolute disparities produced clear changes in the neural responses to unchanged external stimuli, which were well predicted by the measured change in absolute disparity: in 45/53 cases, the neuron maintained a consistent firing pattern with respect to absolute disparity so that the manipulation created no significant change in the absolute disparity preferred by the neuron. No neuron in V1 maintained a consistent relationship with relative disparity. We conclude that the relative disparity signals used in primate depth perception are constructed outside area V1. PMID- 10377366 TI - Direct androgenic regulation of calcitonin gene-related peptide expression in motoneurons of rats with mosaic androgen insensitivity. AB - The spinal nucleus of the bulbocavernosus (SNB) and its target muscles, bulbocavernosus and levator ani (BC/LA), form a sexually dimorphic neuromuscular circuit whose development and maintenance are androgen-dependent. The mechanisms whereby androgen regulates gene expression in the SNB of adult rats are largely unknown, although a retrograde influence from the BC/LA muscles has been suggested to underlie the suppression of calcitonin gene-related peptide (CGRP) expression observed in SNB motoneurons after systemic androgen treatment. A mosaic paradigm was used to determine the site of action of androgen in the regulation of CGRP expression in SNB motoneurons. As a consequence of random X chromosome inactivation, androgenized female rats heterozygous for the tfm androgen receptor (AR) mutation (XwtXtfm-mosaics) express a mosaic of androgen sensitive and androgen-insensitive motoneurons in the SNB, whereas the BC/LA target musculature appears to be uniformly sensitive to androgens. In adult mosaics, testosterone administration resulted in a reduction in the proportion of androgen-sensitive cells expressing CGRP, whereas no such reduction was observed in the androgen-insensitive population, indicating that neuronal AR plays an essential role in the neuromuscular regulation of CGRP expression in these motoneurons. This provides the first in vivo demonstration of AR regulation of gene expression unambiguously localized to a neuronal population. PMID- 10377370 TI - Rat strain differences in the ability to disrupt sensorimotor gating are limited to the dopaminergic system, specific to prepulse inhibition, and unrelated to changes in startle amplitude or nucleus accumbens dopamine receptor sensitivity. AB - Previous studies indicate that a variety of pharmacological agents interfere with the prepulse inhibition of the acoustic startle (PPI) response including phencyclidine (PCP), 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), amphetamine, and apomorphine. Strain differences have been observed in the ability of apomorphine to disrupt PPI, although the degree to which these strain differences occur after administration of nondopaminergic drugs or the degree to which differences can be observed in other models of dopamine (DA) receptor activation has not been elucidated. The present study tested the effects of apomorphine, amphetamine, 8-OH-DPAT, and PCP on PPI in the Sprague Dawley and Wistar rat strains. Because apomorphine disrupts PPI via activation of DA receptors in the nucleus accumbens, apomorphine-induced hyperlocomotion, also a behavioral model of nucleus accumbens DA receptor activation, was measured in both rat strains. Administration of PCP or 8-OH-DPAT attenuated PPI in both strains, whereas apomorphine and amphetamine only attenuated PPI in Wistar rats. The ability of apomorphine to increase motor activity in the absence of a startle eliciting stimulus was similar in the two strains, as was apomorphine-induced hyperlocomotion. A time course analysis of the effects of apomorphine on startle response in Sprague Dawley rats found that changes in the magnitude of PPI followed changes in basic startle amplitude. Similarly, no apomorphine-induced attenuation of PPI was observed in Sprague Dawley rats after 6-OHDA-induced DA receptor supersensitivity in the nucleus accumbens. These data suggest a dissociation between the effects of DA receptor agonists in PPI and other behavioral models of DA receptor activation. PMID- 10377368 TI - Actions of brain-derived neurotrophic factor in slices from rats with spontaneous seizures and mossy fiber sprouting in the dentate gyrus. AB - This study examined the acute actions of brain-derived neurotrophic factor (BDNF) in the rat dentate gyrus after seizures, because previous studies have shown that BDNF has acute effects on dentate granule cell synaptic transmission, and other studies have demonstrated that BDNF expression increases in granule cells after seizures. Pilocarpine-treated rats were studied because they not only have seizures and increased BDNF expression in granule cells, but they also have reorganization of granule cell "mossy fiber" axons. This reorganization, referred to as "sprouting," involves collaterals that grow into novel areas, i.e., the inner molecular layer, where granule cell and interneuron dendrites are located. Thus, this animal model allowed us to address the effects of BDNF in the dentate gyrus after seizures, as well as the actions of BDNF on mossy fiber transmission after reorganization. In slices with sprouting, BDNF bath application enhanced responses recorded in the inner molecular layer to mossy fiber stimulation. Spontaneous bursts of granule cells occurred, and these were apparently generated at the site of the sprouted axon plexus. These effects were not accompanied by major changes in perforant path-evoked responses or paired-pulse inhibition, occurred only after prolonged (30-60 min) exposure to BDNF, and were blocked by K252a. The results suggest a preferential action of BDNF at mossy fiber synapses, even after substantial changes in the dentate gyrus network. Moreover, the results suggest that activation of trkB receptors could contribute to the hyperexcitability observed in animals with sprouting. Because human granule cells also express increased BDNF mRNA after seizures, and sprouting can occur in temporal lobe epileptics, the results may have implications for understanding temporal lobe epilepsy. PMID- 10377369 TI - Spatial attention deficits in patients with acquired or developmental cerebellar abnormality. AB - Recent imaging and clinical studies have challenged the concept that the functional role of the cerebellum is exclusively in the motor domain. We present evidence of slowed covert orienting of visuospatial attention in patients with developmental cerebellar abnormality (patients with autism, a disorder in which at least 90% of all postmortem cases reported to date have Purkinje neuron loss), and in patients with cerebellar damage acquired from tumor or stroke. In spatial cuing tasks, normal control subjects across a wide age range were able to orient attention within 100 msec of an attention-directing cue. Patients with cerebellar damage showed little evidence of having oriented attention after 100 msec but did show the effects of attention orienting after 800-1200 msec. These effects were demonstrated in a task in which results were independent of the motor response. In this task, smaller cerebellar vermal lobules VI-VII (from magnetic resonance imaging) were associated with greater attention-orienting deficits. Although eye movements may also be disrupted in patients with cerebellar damage, abnormal gaze shifting cannot explain the timing and nature of the attention-orienting deficits reported here. These data may be consistent with evidence from animal models that suggest damage to the cerebellum disrupts both the spatial encoding of a location for an attentional shift and the subsequent gaze shift. These data are also consistent with a model of cerebellar function in which the cerebellum supports a broad spectrum of brain systems involved in both nonmotor and motor function. PMID- 10377371 TI - Sensitization to the effects of tumor necrosis factor-alpha: neuroendocrine, central monoamine, and behavioral variations. AB - Consistent with the proposition that cytokines act as immunotransmitters between the immune system and the brain, systemic administration of the proinflammatory cytokine tumor necrosis factor-alpha (TNF-alpha; 1.0-4.0 microg) induced mild illness in CD-1 mice, increased plasma corticosterone concentrations, and altered central norepinephrine, dopamine, and serotonin turnover. The actions of TNF alpha were subject to a time-dependent sensitization effect. After reexposure to a subeffective dose of the cytokine (1.0 microgram) 14-28 d after initial treatment, marked illness was evident (reduced consumption of a palatable substance and diminished activity and social exploration), coupled with an elevation of plasma corticosterone levels. In contrast, cytokine reexposure 1-7 d after initial treatment did not elicit illness, and at the 1 d interval the corticosterone response to the cytokine was reduced. The increase of norepinephrine release within the paraventricular nucleus of the hypothalamus, as reflected by elevated accumulation of 3-methoxy-4-hydroxyphenylglycol, was augmented at the longer reexposure intervals. In contrast, within the central amygdala and the prefrontal cortex TNF-alpha reexposure at the 1 d interval was associated with a pronounced sensitization-like effect, which was not apparent at longer intervals. Evidently, systemic TNF-alpha proactively influences the response to subsequent treatment; however, the nature of the effects (i.e., the behavioral, neuroendocrine, and central transmitter alterations) vary over time after initial cytokine treatment. It is suggested that the sensitization may have important repercussions with respect to cognitive effects of TNF-alpha and may also be relevant to analyses of the neuroprotective or neurodestructive actions of cytokines. PMID- 10377372 TI - An antisense oligonucleotide reverses the footshock-induced expression of fos in the rat medial prefrontal cortex and the subsequent expression of conditioned fear-induced immobility. AB - The immediate-early genes, including c-fos, have been proposed to be involved in learning and memory. In this report, we examine stress-induced Fos-like immunoreactivity (Fos-li) in subregions of the prefrontal cortex during a conditioned fear paradigm. During the acquisition phase, the rats were conditioned to fear a formerly neutral tone by pairing the tone with a mild footshock. The rats were then tested for fearful behavior by reexposure to the tone without additional footshock. During acquisition, Fos-li was increased in the medial prefrontal cortex (infralimbic and prelimbic) but not the anterior cingulate and M1 motor cortex. However, during the extinction phase, no significant increase in Fos-li was observed in any region. These findings indicate that acquisition, but not extinction, of conditioned fear is associated with an increase in Fos-li in subregions of the medial prefrontal cortex. In other animals, an antisense oligonucleotide directed against the c-fos mRNA was injected into the infralimbic/prelimbic cortex 12 or 72 hr before the acquisition session. Antisense treatment given 12, but not 72, hr earlier suppressed Fos production without altering behavior during the acquisition session. Three days after the acquisition session, rats were tested for fearful behavior as before. The antisense oligonucleotide blockade of Fos production during acquisition was associated with a significantly less fearful response during the extinction session. These results support a role for Fos in the medial prefrontal cortex during the acquisition of aversive learning. PMID- 10377373 TI - Activity of neurons in human temporal cortex during identification and memory for names and words. AB - Extracellular recordings of human temporal cortical neuronal activity during identification and memory for object names or words were obtained from 31 neurons at 18 sites in 12 left language dominant patients undergoing left (10) or right (2) awake craniotomy for epilepsy under local anesthesia. Frequency of activity during identification was compared with perceptual controls, that during the encoding phase of recent memory to identification of the same material. Statistically significant changes in one or more temporal epoch (p < 0.005) of one or more comparisons were present for 27 of the 31 neurons in either hemisphere. Few neurons changed activity in the same direction for both words and names. The instruction to retain an item in recent memory changed activity in most neurons from that during identification of the same material, although the items presented were identical and overtly identified in each task. Any individual neuron usually changed activity in one direction for only one task. There are separate, widely distributed neural networks for identification or recent memory for each type of material. The majority of nearby neurons recorded through the same extracellular microelectrode were related to the networks for different tasks. The temporal characteristics of these changes were also investigated; 31% of the changes were "phasic": temporally related to presentation or response to the item. Most of the remaining neuron changes were sustained throughout a task, often for several minutes. These task-specific sustained changes may reflect effects of thalamo-cortical attentional systems. Individual neurons had both sustained and phasic changes to different tasks. PMID- 10377374 TI - Direct evidence for biphasic cAMP responsive element-binding protein phosphorylation during long-term potentiation in the rat dentate gyrus in vivo. AB - Phosphorylation of the transcription factor cAMP responsive element-binding protein (CREB) is thought to play a key role in synaptic plasticity and long-term memory. However, direct evidence for CREB phosphorylation during hippocampal long term potentiation (LTP) in vivo is sparse. Here, we show that, in the intact animal, CREB is rapidly phosphorylated in response to high-frequency stimulation but not low-frequency stimulation of the perforant pathway. CREB phosphorylation occurred in a biphasic manner, with a first peak at 30 min and a second long lasting peak beginning 2 hr after tetanic stimulation and lasting for at least 24 hr. Only stimuli that generated nondecremental LTP promoted a sustained hyperphosphorylation of CREB but not stimuli that produced decremental LTP. CREB phosphorylation was specifically triggered in the dentate gyrus, as well as the CA1, but not the CA3, hippocampal region. Pretreatment with the NMDA receptor antagonist (+)-5-methyl-10,11-dihydro-5H-dibenzo [a,d] cyclohepten-5,10-imine maleate completely prevented activation of CREB. Together, we have resolved the spatial and temporal dynamics of CREB phosphorylation during hippocampal LTP, showing that the transcription factor CREB is specifically recruited at two distinct time points in some forms of hippocampal synaptic plasticity in vivo. PMID- 10377375 TI - Hominids and hybrids: the place of Neanderthals in human evolution. PMID- 10377376 TI - The tangled biology of tau. PMID- 10377377 TI - Coming or going it's another pretty picture for the lambda-Int family album. PMID- 10377378 TI - Augmenting beta receptors in the heart: short-term gains offset by long-term pains? PMID- 10377379 TI - Permanence of brain sex differences and structural plasticity of the adult brain. PMID- 10377380 TI - A method for high-sensitivity peptide sequencing using postsource decay matrix assisted laser desorption ionization mass spectrometry. AB - A method has been developed for de novo peptide sequencing using matrix-assisted laser desorption ionization mass spectrometry. This method will facilitate biological studies that require rapid determination of peptide or protein sequences, e.g., determination of posttranslational modifications, identification of active compounds isolated from combinatorial peptide libraries, and the selective identification of proteins as part of proteome studies. The method involves fast, one-step addition of a sulfonic acid group to the N terminus of tryptic peptides followed by acquisition of postsource decay (PSD) fragment ion spectra. The derivatives are designed to promote efficient charge site-initiated fragmentation of the backbone amide bonds and to selectively enhance the detection of a single fragment ion series that contains the C terminus of the molecule (y-ions). The overall method has been applied to pmol quantities of peptides. The resulting PSD fragment ion spectra often exhibit uninterrupted sequences of 20 or more amino acid residues. However, fragmentation efficiency decreases considerably at amide bonds on the C-terminal side of Pro. The spectra are simple enough that de novo sequence tagging is routine. The technique has been successfully applied to peptide mixtures, to high-mass peptides (up to 3,600 Da) and to the unambiguous identification of proteins isolated from two dimensional gel electrophoresis. The PSD spectra of these derivatized peptides often allow far more selective protein sequence database searches than those obtained from the spectra of native peptides. PMID- 10377381 TI - Purification of transcription cofactor complex CRSP. AB - Transcription of protein coding genes in metazoans involves the concerted action of enhancer binding proteins and the RNA polymerase II apparatus. The cross talk between these two classes of transcription factors is mediated by an elaborate set of cofactor complexes. For the activation of transcription by the promoter specificity protein 1 (Sp1), TATA binding protein-associated factors in the TFIID complex originally were identified as necessary coactivators, but the identity of additional cofactors required for activated transcription was unknown. Recently, we have reported the isolation and properties of a cofactor complex, CRSP (cofactor required for Sp1), which functions in conjunction with the TATA binding protein-associated factors to promote efficient activation of transcription by Sp1. CRSP contains unique subunits as well as polypeptides that are shared with other cofactor complexes. Here, we report a detailed purification protocol for the isolation of CRSP from human HeLa cells. Our purification strategy takes advantage of the ability of CRSP to bind Ni2+-nitrilotriacetic acid-agarose resin as well as other conventional chromatographic resins. We also describe a streamlined purification protocol that allows a more rapid and efficient means to isolate active CRSP. PMID- 10377382 TI - Asymmetric DNA bending in the Cre-loxP site-specific recombination synapse. AB - Cre recombinase catalyzes site-specific recombination between two 34-bp loxP sites in a variety of DNA substrates. At the start of the recombination pathway, the loxP sites are each bound by two recombinase molecules, and synapsis of the sites is mediated by Cre-Cre interactions. We describe the structures of synaptic complexes formed between a symmetrized loxP site and two Cre mutants that are defective in strand cleavage. The DNA in these complexes is bent sharply at a single base pair step at one end of the crossover region in a manner that is atypical of protein-induced DNA bends. A large negative roll (-49 degrees) and a positive tilt (16 degrees) open the major groove toward the center of the synapse and compress the minor groove toward the protein-DNA interface. The bend direction of the site appears to determine which of the two DNA substrate strands will be cleaved and exchanged in the initial stages of the recombination pathway. These results provide a structural basis for the observation that exchange of DNA strands proceeds in a defined order in some tyrosine recombinase systems. The Cre loxS synaptic complex structure supports a model in which synapsis of the loxP sites results in formation of a Holliday junction-like DNA architecture that is maintained through the initial cleavage and strand exchange steps in the site specific recombination pathway. PMID- 10377383 TI - mRNA cap recognition: dominant role of enhanced stacking interactions between methylated bases and protein aromatic side chains. AB - We have determined, by high resolution x-ray analysis, 10 structures comprising the mRNA cap-specific methyltransferase VP39 or specific mutants thereof in the presence of methylated nucleobase analogs (N1-methyladenine, N3-methyladenine, N1 methylcytosine, N3-methylcytosine) and their unmethylated counterparts, or nucleoside N7-methylguanosine. Together with solution affinity studies and previous crystallographic data for N7-methylguanosine and its phosphorylated derivatives, these data demonstrate that only methylated, positively charged bases are bound, indicating that their enhanced stacking with two aromatic side chains of VP39 (Tyr 22 and Phe 180) plays a dominant role in cap recognition. Four key features characterize this stacking interaction: (i) near perfect parallel alignment between the sandwiched methylated bases and aromatic side chains, (ii) substantial areas of overlap in the two-stacked rings, (iii) a 3.4-A interplanar spacing within the overlapping region, and (iv) positive charge in the heterocyclic nucleobase. PMID- 10377384 TI - Chromatin structure analysis of the mouse Xist locus. AB - The Xist gene is expressed exclusively from the inactive X chromosome and plays a central role in regulating X chromosome inactivation. Here we describe experiments aimed at defining the extent of the active chromatin domain of the expressed Xist allele. By using an allele-specific general DNaseI sensitivity assay we show that there is preferential digestion of the expressed allele at sites within the transcribed locus but not in flanking sites located up to 70 kb 5'. A putative proximal boundary for the Xist domain is located within 10 kb upstream of promoter P1. Chromatin in the expressed domain was found to be acetylated at H4 in XX somatic cells but also in XY cells, where Xist is never expressed. A single clear exception to this was the Xist promoter, which is acetylated only in XX cells. These observations concur with the view that H4 acetylation may not be a general marker of active chromatin domains and further support data implicating local promoter acetylation as being of primary functional significance in vivo. PMID- 10377385 TI - Glutamine synthetase inactivation by protein-protein interaction. AB - Glutamine synthetase (GS; EC 6.3.1.2) is the pivotal enzyme of nitrogen metabolism in prokaryotes. Control of bacterial GS activity by reversible adenylylation has provided one of the classical paradigms of signal transduction by cyclic cascades. By contrast, in the present work we show that cyanobacterial GS is controlled by a different mechanism that involves the interaction of two inhibitory polypeptides with the enzyme. Both inactivating factors (IFs), named IF7 and IF17, are required in vivo for complete GS inactivation. Inactive GS-IF7 and GS-IF17 complexes were reconstituted in vitro by using Escherichia coli expressed purified proteins. Our data suggest that control of GS activity is exerted by regulating the levels of IF7 and IF17. PMID- 10377386 TI - Substrate-induced closure of the flap domain in the ternary complex structures provides insights into the mechanism of catalysis by 3-hydroxy-3-methylglutaryl CoA reductase. AB - 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase is the rate-limiting enzyme and the first committed step in the biosynthesis of cholesterol in mammals. We have determined the crystal structures of two nonproductive ternary complexes of HMG-CoA reductase, HMG-CoA/NAD+ and mevalonate/NADH, at 2.8 A resolution. In the structure of the Pseudomonas mevalonii apoenzyme, the last 50 residues of the C terminus (the flap domain), including the catalytic residue His381, were not visible. The structures of the ternary complexes reported here reveal a substrate induced closing of the flap domain that completes the active site and aligns the catalytic histidine proximal to the thioester of HMG-CoA. The structures also present evidence that Lys267 is critically involved in catalysis and provide insights into the catalytic mechanism. PMID- 10377387 TI - Identification of SH2-bbeta as a potent cytoplasmic activator of the tyrosine kinase Janus kinase 2. AB - Janus kinases (JAKs) are cytoplasmic tyrosine kinases critical for signaling by growth hormone (GH) and many other ligands that bind to members of the cytokine receptor superfamily. SH2-Bbeta was previously identified as a JAK2-interacting protein that is tyrosyl phosphorylated in response to GH and other cytokines that activate JAK2. In this study, we examined whether SH2-Bbeta alters the activity of JAK2. SH2-Bbeta, when coexpressed with JAK2, significantly increased the tyrosyl phosphorylation of JAK2 and multiple other cellular proteins and stimulated the kinase activity of JAK2 by approximately 20-fold. Coexpression of SH2-Bbeta with JAK2 dramatically increased tyrosyl phosphorylation of signal transducer and activator of transcription (Stat)5B and Stat3, physiological substrates of JAK2. SH2-Bbeta(R555E) with a defective Src homology 2 domain was unable to stimulate JAK2 and JAK2-mediated tyrosyl phosphorylation of Stat5B and Stat3. More importantly, SH2-Bbeta enhanced GH-induced tyrosyl phosphorylation of endogenous JAK2 and ligand-induced tyrosyl phosphorylation of Stat5B by endogenous JAK2. In contrast, SH2-Bbeta did not potentiate the activation of other tyrosine kinases including the receptors for platelet-derived growth factor, epidermal growth factor, or nerve growth factor (TrkA), tyrosine kinases that also bind SH2-Bbeta. These data demonstrate that SH2-Bbeta is a potent cytoplasmic activator of JAK2 and is thereby expected to be an important cellular regulator of signaling by GH and other hormones and cytokines that activate JAK2. PMID- 10377388 TI - Comparison of the intracellular signaling responses by three chimeric fibroblast growth factor receptors in PC12 cells. AB - Stably transfected PC12 cell lines expressing similar amounts of chimeric receptors composed of the extracellular domain of the human platelet-derived growth factor (PDGF)beta receptor and the transmembrane and intracellular domains of the fibroblast growth factor receptors (FGFRs) 1, 3, and 4 undergo ligand induced differentiation. The FGFR1 chimera (PFR1) is the most potent of the three, and PFR4 requires more frequent (every 24 hr) addition of ligand to maintain the response. Both PFR1 and -3 also show significant ligand-independent autophosphorylation but PFR4 does not. All of the chimeras activated phospholipase Cgamma, Shc, FGFR substrate (FRS)2, and the mitogen-activated protein kinases, ERK1 and 2. PFR4 was moderately weaker in stimulating these effects as well; PFR1 and -3 were comparable. None of the chimeras induced Sos association or were coprecipitated with Shc. Cotransfection of a dominant negative Shc derivative, with tyrosine at 239, 240, and 317 replaced with phenylalanine, in the PFR-expressing cells was without effect on PDGF-induced neurite outgrowth. The same derivative substantially inhibited the response of these cells to NGF. These results indicate that FGFR1, 3, and 4 (i) are capable of signaling in a similar fashion; (ii) primarily use FRS2 and, perhaps, PLCgamma; and (iii) do not utilize Shc. The results also suggest that the principal difference between FGFR1, 3, and 4 is in the strength of the tyrosine kinase activity and that qualitative differences in signaling capacity are likely to be less important. PMID- 10377389 TI - Heat-inactivated proteins are rescued by the DnaK.J-GrpE set and ClpB chaperones. AB - Functional chaperone cooperation between Hsp70 (DnaK) and Hsp104 (ClpB) was demonstrated in vitro. In a eubacterium Thermus thermophilus, DnaK and DnaJ exist as a stable trigonal ring complex (TDnaK.J complex) and the dnaK gene cluster contains a clpB gene. When substrate proteins were heated at high temperature, none of the chaperones protected them from heat inactivation, but the TDnaK.J complex could suppress the aggregation of proteins in an ATP- and TGrpE-dependent manner. Subsequent incubation of these heated preparations at moderate temperature after addition of TClpB resulted in the efficient reactivation of the proteins. Reactivation was also observed, even though the yield was low, if the substrate protein alone was heated and incubated at moderate temperature with the TDnaK.J complex, TGrpE, TClpB, and ATP. Thus, all these components were necessary for the reactivation. Further, we found that TGroEL/ES could not substitute TClpB. PMID- 10377390 TI - Bypassing the periplasm: reconstitution of the AcrAB multidrug efflux pump of Escherichia coli. AB - AcrAB is a constitutively expressed, major multidrug efflux system of Escherichia coli. We have purified the cytoplasmic membrane component, AcrB, to near homogeneity, and reconstituted the protein into proteoliposomes. In the presence of DeltapH (outside acid), the protein catalyzed the extrusion of fluorescent phospholipids, which were then trapped by protein-free acceptor vesicles. Known substrates of AcrAB, such as bile acids, erythromycin, and cloxacillin, inhibited this activity. Addition of various drugs to AcrB-containing proteoliposomes, in the presence of DeltapH (inside acid) resulted in proton efflux, suggesting that AcrB is a proton antiporter. Interestingly, fluorescent lipid extrusion was accelerated strongly by the periplasmic protein AcrA in the presence of Mg2+, and at pH 5.0 AcrA alone produced a slow mixing of lipids of different vesicles, without causing the mixing of intravesicular material. These results suggest that AcrA brings two membranes together, and under certain conditions may even cause the fusion of at least the outer leaflets of the membranes, contributing to the ability of the AcrAB-TolC system to pump drugs out directly into the medium. PMID- 10377391 TI - Poisoning of human DNA topoisomerase I by ecteinascidin 743, an anticancer drug that selectively alkylates DNA in the minor groove. AB - Ecteinascidin 743 (Et743, National Service Center 648766) is a potent antitumor agent from the Caribbean tunicate Ecteinascidia turbinata. Although Et743 is presently in clinical trials for human cancers, the mechanisms of antitumor activity of Et743 have not been elucidated. Et743 can alkylate selectively guanine N2 from the DNA minor groove, and this alkylation is reversed by DNA denaturation. Thus, Et743 differs from other DNA alkylating agents presently in the clinic (by both its biochemical activities and its profile of antitumor activity in preclinical models). In this study, we investigated cellular proteins that can bind to DNA alkylated by Et743. By using an oligonucleotide containing high-affinity Et743 binding sites and nuclear extracts from human leukemia CEM cells, we purified a 100-kDa protein as a cellular target of Et743 and identified it as topoisomerase I (top1). Purified top1 was then tested and found to produce cleavage complexes in the presence of Et743, whereas topoisomerase II had no effect. DNA alkylation was essential for the formation of top1-mediated cleavage complexes by Et743, and the distribution of the drug-induced top1 sites was different for Et743 and camptothecin. top1-DNA complexes were also detected in Et743-treated CEM cells by using cesium chloride gradient centrifugation followed by top1 immunoblotting. These data indicate that DNA minor groove alkylation by Et743 induces top1-mediated protein-linked DNA breaks and that top1 is a target for Et743 in vitro and in vivo. PMID- 10377392 TI - An inactive open complex mediated by an UP element at Escherichia coli promoters. AB - A specific interaction between the alpha subunit of RNA polymerase and an A+T rich upstream sequence (UP element) stimulates transcription at some promoters in Escherichia coli. We found that RNA polymerase formed a heparin-resistant nonproductive initiation complex at the malT promoter which has an A+T-rich upstream sequence that begins 9 bp upstream of the -35 region. Substitution of other sequences for the A+T-rich sequence eliminated both the formation of heparin-resistant complexes and alpha binding to the malT promoter. A 5-bp deletion between the A+T-rich sequence and the -35 region increased promoter activity. The UP element derived from the rrnB P1 promoter stimulated transcription of the malT core promoter when placed 4 bp upstream from the malT 35 region, but insertion of an additional 4 bp between the rrnB P1 UP element and the -35 element eliminated transcription activity without eliminating heparin resistant complex formation. Similar UP element effects were observed in hybrids with the lac core promoter, even though the region around the transcription start site was melted in both productive and nonproductive complexes. We conclude that UP elements can mediate the formation of both productive and nonproductive open complexes, depending on their location with respect to the core promoter. PMID- 10377393 TI - Host-cell positive transcription elongation factor b kinase activity is essential and limiting for HIV type 1 replication. AB - HIV-1 gene expression and viral replication require the viral transactivator protein Tat. The RNA polymerase II transcriptional elongation factor P-TEFb (cyclin-dependent kinase 9/cyclin T) is a cellular protein kinase that has recently been shown to be a key component of the Tat-transactivation process. For this report, we studied the requirement for P-TEFb in HIV-1 infection, and we now show that P-TEFb is both essential and limiting for HIV-1 replication. Attenuation of P-TEFb kinase activity either by expression of a dominant-negative cyclin-dependent kinase 9 transgene or through the use of small-molecule inhibitors suppresses HIV-1 gene expression and HIV-1 replication. Inhibition of HIV-1 replication is affected in a manner consistent with a direct and specific effect on P-TEFb and the known functional role of P-TEFb in Tat-activated transcription. Tat-activated expression of HIV-1 genes seems uniquely dependent on P-TEFb, as inhibition of P-TEFb activity and HIV-1 replication can be achieved without compromising cell viability or RNA polymerase II-dependent cellular gene transcription. Selective inhibition of the P-TEFb kinase may therefore provide a novel approach for developing chemotherapeutic agents against HIV-1. PMID- 10377394 TI - NFAT5, a constitutively nuclear NFAT protein that does not cooperate with Fos and Jun. AB - NFAT transcription factors are related to NF-kappaB/Rel proteins and form cooperative complexes with Fos and Jun on DNA. We have identified an NFAT-related protein, NFAT5, which differs from the conventional NFAT proteins NFAT1-4 in its structure, DNA binding, and regulation. NFAT5 contains a NFAT-like Rel homology domain, conserves the DNA contact residues of NFAT1-4, and binds DNA sequences similar to those found in the regulatory regions of well-characterized NFAT dependent genes. However, it lacks the majority of Fos/Jun contact residues and does not bind cooperatively with Fos and Jun to DNA. Unlike NFAT1-4, whose nuclear import is tightly regulated by calcineurin-mediated dephosphorylation, NFAT5 is a constitutively nuclear phosphoprotein regardless of calcineurin activation. These features suggest that unlike the conventional NFAT proteins, NFAT1-4, which activate gene transcription by integrating inputs from calcium/calcineurin and protein kinase C/mitogen-activated protein kinase signaling pathways, NFAT5 participates in as-yet-unidentified signaling pathways in diverse immune and nonimmune cells. PMID- 10377395 TI - Identification of human prostaglandin E synthase: a microsomal, glutathione dependent, inducible enzyme, constituting a potential novel drug target. AB - Human prostaglandin (PG) E synthase (EC 5.3.99.3) is a member of a recently recognized protein superfamily consisting of membrane associated proteins involved in eicosanoid and glutathione metabolism (the MAPEG family). Previous designations of the protein are PIG12 and MGST1-L1. PGE synthase was expressed in Escherichia coli, and both cytosolic and membrane fractions were prepared. Western blot analysis specifically detected a 15- to 16-kDa protein in the membrane fraction. Both fractions were incubated with prostaglandin H2 in the presence or absence of reduced glutathione. The membrane but not the cytosolic fraction was found to possess high glutathione-dependent PGE synthase activity (0.25 micromol/min/mg). The human tissue distribution was analyzed by Northern blot analysis. High expression of PGE synthase mRNA was detected in A549 and HeLa cancer cell lines. Intermediate level of expression was demonstrated in placenta, prostate, testis, mammary gland, and bladder whereas low mRNA expression was observed in several other tissues. A549 cells have been used as a model system to study cyclooxygenase-2 induction by IL-1beta. If A549 cells were grown in the presence of IL-1beta, a significant induction of the PGE synthase was observed by Western blot analysis. Also, Western blot analysis specifically detected a 16-kDa protein in sheep seminal vesicles. In summary, we have identified a human membrane bound PGE synthase. The enzyme activity is glutathione-dependent, and the protein expression is induced by the proinflammatory cytokine IL-1beta. PGE synthase is a potential novel target for drug development. PMID- 10377396 TI - Purification of the yeast U4/U6.U5 small nuclear ribonucleoprotein particle and identification of its proteins. AB - The yeast U4/U6.U5 pre-mRNA splicing small nuclear ribonucleoprotein (snRNP) is a 25S small nuclear ribonucleoprotein particle similar in size, composition, and morphology to its counterpart in human cells. The yeast U4/U6.U5 snRNP complex has been purified to near homogeneity by affinity chromatography and preparative glycerol gradient sedimentation. We show that there are at least 24 proteins stably associated with this particle and performed mass spectrometry microsequencing to determine their identities. In addition to the seven canonical core Sm proteins, there are a set of U6 snRNP specific Sm proteins, eight previously described U4/U6.U5 snRNP proteins, and four novel proteins. Two of the novel proteins have likely RNA binding properties, one has been implicated in the cell cycle, and one has no identifiable sequence homologues or functional motifs. The purification of the low abundance U4/U6.U5 snRNP from yeast and the powerful sequencing methodologies using small amounts of protein make possible the rapid identification of novel and previously unidentified components of large, low abundance macromolecular machines from any genetically manipulable organism. PMID- 10377397 TI - The alpha-helix folds on the millisecond time scale. AB - It has long been believed that nucleation of the alpha-helix is a very fast reaction, occurring in around 10(-7) s. We show here that helix nucleation, in fact, takes place on the millisecond time scale. The rate of alpha-helix nucleation in two polyalanine-based peptides and in lysine and glutamic acid homopolymers was measured directly by stopped-flow deep UV CD with synchrotron radiation as the light source. Synchrotron radiation CD gives far superior signal to noise than a conventional instrument. The 16-aa AK peptide folds with first order kinetics and a rate constant of 15 s-1 at 0 degrees C. The rate-determining step is presumably the initiation of a new helix, which occurs at least 10(5) times slower than expected. Helix folding occurs in at least two steps on the millisecond time scale for the longer peptides, with a transient overshoot of helix content significantly greater than at equilibrium, similar to that seen in the folding of several proteins. We suggest that the overshoot is caused by the formation of a single long helix followed by its breakage into the two or more helices present at equilibrium. PMID- 10377398 TI - cDNA cloning of cholesterol 24-hydroxylase, a mediator of cholesterol homeostasis in the brain. AB - The turnover of cholesterol in the brain is thought to occur via conversion of excess cholesterol into 24S-hydroxycholesterol, an oxysterol that is readily secreted from the central nervous system into the plasma. To gain molecular insight into this pathway of cholesterol metabolism, we used expression cloning to isolate cDNAs that encode murine and human cholesterol 24-hydroxylases. DNA sequence analysis indicates that both proteins are localized to the endoplasmic reticulum, share 95% identity, and represent a new cytochrome P450 subfamily (CYP46). When transfected into cultured cells, the cDNAs produce an enzymatic activity that converts cholesterol into 24S-hydroxycholesterol, and to a lesser extent, 25-hydroxycholesterol. The cholesterol 24-hydroxylase gene contains 15 exons and is located on human chromosome 14q32.1. Cholesterol 24-hydroxylase is expressed predominantly in the brain as judged by RNA and protein blotting. In situ mRNA hybridization and immunohistochemistry localize the expression of this P450 to neurons in multiple subregions of the brain. The concentrations of 24S hydroxycholesterol in serum are low in newborn mice, reach a peak between postnatal days 12 and 15, and thereafter decline to baseline levels. In contrast, cholesterol 24-hydroxylase protein is first detected in the brain of mice at birth and continues to accumulate with age. We conclude that the cloned cDNAs encode cholesterol 24-hydroxylases that synthesize oxysterols in neurons of the brain and that secretion of 24S-hydroxycholesterol from this tissue in the mouse is developmentally regulated. PMID- 10377399 TI - Electrically driven motor in the outer hair cell: effect of a mechanical constraint. AB - The outer hair cell has a unique voltage-dependent motility associated with charge transfer across the plasma membrane. To examine mechanical changes in the membrane that are coupled with such charge movements, we digested the undercoating of the membrane with trypsin. We inflated the cell into a sphere and constrained the surface area by not allowing volume changes. We found that this constraint on the membrane area sharply reduced motor-associated charge movement across the membrane, demonstrating that charge transfer is directly coupled with membrane area change. This electromechanical coupling in the plasma membrane must be the key element for the motile mechanism of the outer hair cell. PMID- 10377401 TI - A single amino acid near the C terminus of the synaptosomeassociated protein of 25 kDa (SNAP-25) is essential for exocytosis in chromaffin cells. AB - Amperometry in chromaffin cells expressing green fluorescent protein (GFP) fused to synaptosome-associated protein of 25 kDa (SNAP-25) have been used to test the involvement of single amino acids in exocytotic function, overcoming some of the limitations of studies based on Botulinum neurotoxin cleavage, as this occurs at defined sites of the protein. Constructs containing either the whole SNAP-25 polypeptide or several deleted forms lacking its C-terminal domain were heavily overexpressed in transfected cells. All GFP-fusions were located in both the cytoplasm and the plasma membrane. Although a construct containing complete SNAP 25 sustained normal secretion, removal of four or more amino acids of its C terminus greatly altered the overall rate and extent of exocytosis. Further mutational analysis proved that Leu203, the fourth residue from the C terminus, is critical for secretion. Kinetics of single granule fusions from cells expressing truncated forms showed slow onset and decay times when compared with control cells expressing full SNAP-25. Thus, these data provide direct evidence for the involvement of a specific residue of SNAP-25 in exocytosis and show that overexpression of GFP-SNAP contructs combined with single vesicle fusion measurements constitutes a powerful approach to dissect the structural elements playing a role in individual steps of the exocytotic cascade. PMID- 10377400 TI - The active form of the steroidogenic acute regulatory protein, StAR, appears to be a molten globule. AB - The steroidogenic acute regulatory protein (StAR) increases the movement of cholesterol from the outer to the inner membrane of adrenal and gonadal mitochondria, thus providing the substrate for steroid hormone biosynthesis. Deletion of 62 amino-terminal aa produces a cytoplasmic form of StAR (N-62 StAR) that lacks the mitochondrial leader sequence but retains full activity and appears to act at the outer mitochondrial membrane. At neutral pH the native state of bacterially expressed N-62 StAR protein displays cooperative unfolding under the influence of urea with DeltaGH2O = -4.1 kcal/mol, and it remains correctly folded down to pH 4. Limited proteolysis at different pHs shows that the biologically essential C-terminal region is accessible to solvent, and that the N-terminal domain is compact at pH 8 and partially unfolds below pH 4. Secondary structural analysis of CD curves suggests that the unfolding may coincide with an increase in alpha-helical character at pH 3.5. Fluorescence spectroscopy at pH 3-8 and at 0-6 M urea is consistent with two distinct domains, a compact N-terminal domain containing tryptophans 96 and 147 and a more solvent accessible C-terminal domain containing tryptophans 241 and 250. These observations suggest that StAR forms a molten globule structure at pH 3.5-4.0. As the mitochondrial proton pump results in an electrochemical gradient, and as StAR must unfold during mitochondrial entry, StAR probably undergoes a similar conformational shift to an extended structure while interacting with the mitochondrial outer membrane, allowing this apparent molten globule form to act as an on/off switch for cholesterol entry into the mitochondria. PMID- 10377402 TI - Passive entry of a DNA molecule into a small pore. AB - I consider a vesicle with an open pore of small radius rp, exposed to a DNA solution. The crucial moment is the entry, when a chain end faces the pore and enters it. I discuss qualitatively the following three characteristic times: (i) the duration of the entry of one chain end (defining the minimum lifetime of the pore) taue approximately 10(-4) sec, (ii) the transfection time taut (the time required to be sure that one chain has gone in) taut approximately hours, and (iii) the sliding time tauS (the time between entry of one end and entry of the other end) approximately 1 sec. A fortunate feature is that sliding may proceed even if the pore tends to close itself after entry. PMID- 10377403 TI - Hepatic differentiation induced by oncostatin M attenuates fetal liver hematopoiesis. AB - Embryonic liver is a transient site for definitive hematopoiesis. Along with maturation of the bone marrow and spleen, hematopoietic cells relocate from the liver to their final destinations while the liver starts organizing its own structure and develops numerous metabolic functions toward adult. Recently, it was demonstrated that the signal exerted by oncostatin M (OSM) through gp130 plays a pivotal role in the maturation process of the liver both in vitro and in vivo. However, the molecular basis underlying the termination of embryonic hematopoiesis remains unknown. In this study, we report that primary culture of fetal hepatic cells from embryonic day 14.5 murine embryos supported expansion of blood cells from Lin-Sca-1(+)c-Kit+ cells, giving rise to myeloid, lymphoid, and erythroid lineages. Of interest, promotion of hepatic development by OSM and glucocorticoid strongly suppressed in vitro hematopoiesis. Consistent with these results, hepatic culture from the embryonic day 18.5 liver no longer supported hematopoiesis. These data together with the previous observations suggest that the signals exerted by OSM and glucocorticoid induce hepatic differentiation, which in turn terminate embryonic hematopoiesis and promote relocation of hematopoietic cells. PMID- 10377404 TI - Receptor recruitment: a mechanism for interactions between G protein-coupled receptors. AB - There is a great deal of evidence for synergistic interactions between G protein coupled signal transduction pathways in various tissues. As two specific examples, the potent effects of the biogenic amines norepinephrine and dopamine on sodium transporters and natriuresis can be modulated by neuropeptide Y and atrial natriuretic peptide, respectively. Here, we report, using a renal epithelial cell line, that both types of modulation involve recruitment of receptors from the interior of the cell to the plasma membrane. The results indicate that recruitment of G protein-coupled receptors may be a ubiquitous mechanism for receptor sensitization and may play a role in the modulation of signal transduction comparable to that of the well established phenomenon of receptor endocytosis and desensitization. PMID- 10377405 TI - Separation of C/EBPalpha-mediated proliferation arrest and differentiation pathways. AB - Cell proliferation and terminal differentiation are mutually exclusive in most cell lineages. The b-zip transcription factor CCAAT/enhancer-binding protein alpha (C/EBPalpha) induces proliferation arrest and differentiation in many cell types, suggesting that both activities are linked. Here we show that C/EBPalpha mediated proliferation arrest and differentiation pathways can be separated by the E7 oncoprotein of the "high-risk" human papilloma virus 16. The E7 oncoprotein overrides C/EBPalpha-mediated cell cycle withdrawal without compromising the transactivation activity of C/EBPalpha or its ability to participate in differentiation. Uncoupling of both pathways depends on the casein kinase II site of the oncoprotein but not on its ability to neutralize pocket proteins or the cyclin-dependent kinase inhibitor protein p21. Our results suggest a bifurcation of C/EBPalpha-mediated proliferation arrest and differentiation pathways. PMID- 10377406 TI - A functional bone morphogenetic protein system in the ovary. AB - Bone morphogenetic proteins (BMPs) comprise a large group of polypeptides in the transforming growth factor beta superfamily with essential physiological functions in morphogenesis and organogenesis in both vertebrates and invertebrates. At present, the role of BMPs in the reproductive system of any species is poorly understood. Here, we have established the existence of a functional BMP system in the ovary, replete with ligand, receptor, and novel cellular functions. In situ hybridization histochemistry identified strong mRNA labeling for BMP-4 and -7 in the theca cells and BMP receptor types IA, IB, and II in the granulosa cells and oocytes of most follicles in ovaries of normal cycling rats. To explore the paracrine function of this BMP system, we examined the effects of recombinant BMP-4 and -7 on FSH (follicle-stimulating hormone) induced rat granulosa cytodifferentiation in serum-free medium. Both BMP-4 and -7 regulated FSH action in positive and negative ways. Specifically, physiological concentrations of the BMPs enhanced and attenuated the stimulatory action of FSH on estradiol and progesterone production, respectively. These effects were dose- and time-dependent. Furthermore, the BMPs increased granulosa cell sensitivity to FSH. Thus, BMPs have now been identified as molecules that differentially regulate FSH-dependent estradiol and progesterone production in a way that reflects steroidogenesis during the normal estrous cycle. As such, it can be hypothesized that BMPs might be the long-sought "luteinization inhibitor" in Graafian follicles during their growth and development. PMID- 10377407 TI - Genetic dissection of the budding yeast Arp2/3 complex: a comparison of the in vivo and structural roles of individual subunits. AB - In previous work, we identified the yeast Arp2/3 complex, which localizes to cortical actin patches and is required for their motility and integrity in vivo. This complex contains proteins homologous to each subunit of the Acanthamoeba and human Arp2/3 complex except for a 40-kDa subunit (p40), which was missing from the purified yeast complex. Here, we demonstrate by using immunoprecipitation and gel-filtration analysis that Arc40p, the homolog of p40 identified from the yeast genome database, associates with the yeast Arp2/3 complex. We have carried out gene disruptions of each subunit of the yeast Arp2/3 complex to study each subunit's role in the function of the complex. Surprisingly, we find that only ARC40 is fully essential for cell viability. Strains lacking each of the other subunits exhibit varying degrees of defects in cell growth and viability and in assembly and polarization of cortical actin patches. We have also examined each subunit's role in maintaining the structural integrity of the Arp2/3 complex. Arp2p, Arp3p, and Arc40p fall into the monomer pool in Deltaarc19 and Deltaarc35 cells, suggesting that Arc19p and Arc35p are the central scaffolding components of the complex. Arp2p and Arp3p do not have major roles in maintaining complex integrity, and Arc15p is required for association of Arp2p and Arc40p, but not other subunits, with the complex. These results provide evidence that each subunit contributes differently to the assembly and function of the Arp2/3 complex. PMID- 10377408 TI - Osteoblast-specific gene expression after transplantation of marrow cells: implications for skeletal gene therapy. AB - Somatic gene therapies require targeted transfer of the therapeutic gene(s) into stem cells that proliferate and then differentiate and express the gene in a tissue-restricted manner. We have developed an approach for gene therapy using marrow cells that takes advantage of the osteoblast specificity of the osteocalcin promoter to confine expression of chimeric genes to bone. Adherent marrow cells, carrying a reporter gene [chloramphenicol acetyltransferase (CAT)] under the control of a 1.7-kilobase rat osteocalcin gene promoter, were expanded ex vivo. After transplantation by intravenous infusion, engrafted donor cells in recipient mice were detected by the presence of the transgene in a broad spectrum of tissues. However, expression of the transgene was restricted to osteoblasts and osteocytes, as established by biochemical analysis of CAT activity and immunohistochemical analysis of CAT expression at the single cell level. Our data indicate that donor cells achieved long-term engraftment in various tissues of the recipients and that the CAT gene under control of the osteocalcin promoter is expressed specifically in bone. Thus, transplantation of multipotential marrow cells containing the osteocalcin promoter-controlled transgene provides an efficacious approach to deliver therapeutic gene expression to osteoblasts for treatment of bone disorders or tumor metastasis to the skeleton. PMID- 10377409 TI - Ezrin, a plasma membrane-microfilament linker, signals cell survival through the phosphatidylinositol 3-kinase/Akt pathway. AB - ERM (Ezrin-Radixin-Moesin) proteins function as plasma membrane-actin cytoskeleton linkers and participate in the formation of specialized domains of the plasma membrane. We have investigated ezrin function in tubulogenesis of a kidney-derived epithelial cell line, LLC-PK1. Here we show that cells overproducing a mutant form of ezrin in which Tyr-353 was changed to a phenylalanine (Y353F) undergo apoptosis when assayed for tubulogenesis. While investigating the mechanism responsible for this apoptosis, we found that ezrin interacts with p85, the regulatory subunit of phosphatidylinositol 3-kinase (PI 3 kinase). Two distinct sites of ezrin are involved in this interaction, the amino terminal domain containing the first 309 aa and the phosphorylated Tyr-353 residue, which binds to the carboxyl-terminal SH2 domain of p85. Cells producing Y353F ezrin are defective in activation of the protein kinase Akt, a downstream target of PI 3-kinase that protects cells against apoptosis. Furthermore, the apoptotic phenotype of these cells is rescued by production of a constitutively activated form of PI 3-kinase. Taken together, these results establish a novel function for ezrin in determining survival of epithelial cells by activating the PI 3-kinase/Akt pathway. PMID- 10377410 TI - Cdc20 associates with the kinase aurora2/Aik. AB - Cdc20/fizzy family proteins are involved in activation of the anaphase-promoting complex/cyclosome, which catalyzes the ubiquitin-dependent proteolysis of cell cycle regulatory proteins such as anaphase inhibitors and mitotic cyclins, leading to chromosome segregation and exit from mitosis. Previous work has shown that human Cdc20 (hCdc20/p55CDC) associates with one or more kinases. We report here that Cdc20-associated myelin basic protein kinase activity peaks sharply in early M phase (embryonic cells) or in G2 phase (somatic cells). In HeLa cells, Cdc20 is associated with the kinase aurora2/Aik. Aurora2/Aik is a member of the aurora/Ipl1 family of kinases that, like Cdc20, previously has been shown to be localized at mitotic spindle poles and is involved in regulating chromosome segregation and maintaining genomic stability. The demonstration that Cdc20 is associated with aurora2/Aik suggests that some function of Cdc20 is carried out or regulated through its association with aurora2/Aik. PMID- 10377411 TI - The intracellular parasite Theileria parva protects infected T cells from apoptosis. AB - Parasites have evolved a plethora of strategies to ensure their survival. The intracellular parasite Theileria parva secures its propagation and spreads through the infected animal by infecting and transforming T cells, inducing their continuous proliferation and rendering them metastatic. In previous work, we have shown that the parasite induces constitutive activation of the transcription factor NF-kappaB, by inducing the constitutive degradation of its cytoplasmic inhibitors. The biological significance of NF-kappaB activation in T. parva infected cells, however, has not yet been defined. Cells that have been transformed by viruses or oncogenes can persist only if they manage to avoid destruction by the apoptotic mechanisms that are activated on transformation and that contribute to maintain cellular homeostasis. We now demonstrate that parasite-induced NF-kappaB activation plays a crucial role in the survival of T. parva-transformed T cells by conveying protection against an apoptotic signal that accompanies parasite-mediated transformation. Consequently, inhibition of NF kappaB nuclear translocation and the expression of dominant negative mutant forms of components of the NF-kappaB activation pathway, such as IkappaBalpha or p65, prompt rapid apoptosis of T. parva-transformed T cells. Our findings offer important insights into parasite survival strategies and demonstrate that parasite-induced constitutive NF-kappaB activation is an essential step in maintaining the transformed phenotype of the infected cells. PMID- 10377412 TI - Reprogramming of intestinal differentiation and intercalary regeneration in Cdx2 mutant mice. AB - The homeobox gene Cdx2, a homologue of the Drosophila gene caudal, has been implicated in the control of cell differentiation in the intestinal epithelium. Recently, we showed that mice in which one allele of the Cdx2 gene had been inactivated by homologous recombination developed multiple intestinal polyp-like lesions that did not express Cdx2 and that contained areas of squamous metaplasia in the form of keratinizing stratified squamous epithelium, similar to that occurring in the mouse esophagus and forestomach. We have now examined colonic lesions from 98 Cdx2+/- mice and report that the lesions are composed of heterotopic stomach and small intestinal mucosa. We conclude that Cdx2 directs endodermal differentiation toward a caudal phenotype and that haploinsufficient levels of expression in the developing distal intestine lead to homeotic transformation to a more rostral endodermal phenotype, such as forestomach epithelium that does not express Cdx2 during normal development. Intercalary growth (epimorphic regeneration), which previously has never been described in mammals, then occurs, resulting in the ordered "filling in" of tissue types at the discontinuity between the gastric and colonic epithelia. This intercalary growth in a restricted space results in the formation of the polypoid lesions observed. PMID- 10377413 TI - Normal growth and development in the absence of hepatic insulin-like growth factor I. AB - The somatomedin hypothesis proposed that insulin-like growth factor I (IGF-I) was a hepatically derived circulating mediator of growth hormone and is a crucial factor for postnatal growth and development. To reassess this hypothesis, we have used the Cre/loxP recombination system to delete the igf1 gene exclusively in the liver. igf1 gene deletion in the liver abrogated expression of igf1 mRNA and caused a dramatic reduction in circulating IGF-I levels. However, growth as determined by body weight, body length, and femoral length did not differ from wild-type littermates. Although our model proves that hepatic IGF-I is indeed the major contributor to circulating IGF-I levels in mice it challenges the concept that circulating IGF-I is crucial for normal postnatal growth. Rather, our model provides direct evidence for the importance of the autocrine/paracrine role of IGF-I. PMID- 10377414 TI - Branching morphogenesis independent of mesenchymal-epithelial contact in the developing kidney. AB - Whether mesenchymal-epithelial interactions leading to branching morphogenesis in developing epithelial tissues such as the kidney require direct cell-cell contact or are due to soluble mediators elaborated by the inducing tissue has been the subject of much debate. Here we demonstrate that ureteric bud (UB) epithelium, from which the kidney collecting system and upper urinary tract are derived, can undergo impressive three-dimensional branching morphogenesis when cultured in the appropriate extracellular matrix context in the absence of direct contact with mesenchymal tissue, indicating that the program for branching morphogenesis is inherent to the UB. Both a soluble factor in BSN cell-conditioned medium (BSN-CM) derived from an immortalized cell line thought to originate in the early metanephric mesenchyme and glial cell line-derived neurotrophic factor (GDNF) were required for early and later events in branching morphogenesis. In the absence of BSN-CM, the isolated UB did not survive; a similar result was obtained in the presence of neutralizing antibodies against glial cell line-derived neurotrophic factor. Preliminary analysis of key activity present in BSN-CM indicates that it is a heat-sensitive, heparin-binding factor with a probable molecular mass greater than 100 kDa. When the in vitro cultured UB was recombined with freshly isolated metanephric mesenchyme, nephric units were induced in the mesenchyme, and the UB branches underwent elongation. Our data suggest that, although UB branching morphogenesis per se does not require direct mesenchymal contact, such contact may play a key role in regulating branch elongation and establishing the pattern of branching. The results also suggest an approach to in vitro engineering of nephron. PMID- 10377415 TI - Selective activation of the versican promoter by epithelial- mesenchymal interactions during hair follicle development. AB - Interaction between the epithelium and the mesenchyme is an essential feature of organogenesis, including hair follicle formation. The dermal papilla (DP), a dense aggregate of specialized dermis-derived stromal cells located at the bottom of the follicle, is a major component of hair that signals the follicular epithelial cells to prolong the hair growth process. However, little is known about DP-specific gene activation with regard to hair induction. In this study we demonstrate that a short fragment (839 bp) of the human versican (a core protein of one of the matrix chondroitin sulfate proteoglycans) promoter is sufficient to activate lacZ reporter gene expression in the DP of postnatal transgenic mice and also in the condensed mesenchyme (the origin of the DP) beneath the hair placode during hair follicle embryogenesis. Using the same versican promoter with green fluorescent protein (GFP), large numbers of fresh pelage DP cells were isolated from newborn transgenic skin by high-speed cell sorting. These GFP-positive DP cells showed abundant versican mRNA, confirming that the reporter molecules reflected endogenous versican gene expression. These sorted GFP-positive cells showed DP-like morphology in culture, but both GFP and versican expression was lost during primary culture. In vivo hair growth assays showed that GFP-positive cells could induce hair when grafted with epithelial cells, whereas GFP-negative cells grafted with epithelium or GFP-positive cells alone did not. These results suggest that versican may play an essential role both in mesenchymal condensation and in hair induction. PMID- 10377416 TI - MADS-box genes reveal that gnetophytes are more closely related to conifers than to flowering plants. AB - The evolutionary origin of the angiosperms (flowering plants sensu stricto) is still enigmatic. Answers to the question of angiosperm origins are intimately connected to the identification of their sister group among extinct and extant taxa. Most phylogenetic analyses based on morphological data agree that among the groups of extant seed plants, the gnetophytes are the sister group of the angiosperms. According to this view, angiosperms and gnetophytes are the only extant members of a clade called "anthophytes" to emphasize their shared possession of flower-like reproductive structures. However, most phylogeny reconstructions based on molecular data so far did not support an anthophyte clade, but also could not clarify the case because support for alternative groupings has been weak or controversial. We have isolated 13 different homologs of MADS-type floral homeotic genes from the gnetophyte Gnetum gnemon. Five of these genes fall into monophyletic gene clades also comprising putatively orthologous genes from flowering plants and conifers, among them orthologs of floral homeotic B and C function genes. Within these clades the Gnetum genes always form distinct subclades together with the respective conifer genes, to the exclusion of the angiosperm genes. This provides strong molecular evidence for a sister-group relationship between gnetophytes and conifers, which is in contradiction to widely accepted interpretations of morphological data for almost a century. Our phylogeny reconstructions and the outcome of expression studies suggest that complex features such as flower-like reproductive structures and double-fertilization arose independently in gnetophytes and angiosperms. PMID- 10377417 TI - Mutation, recombination, and incipient speciation of bacteria in the laboratory. AB - Mutations in the DNA mismatch repair system increase mutation and recombination. They may thereby promote the genetic divergence that underlies speciation, after which the reacquisition of a functional repair system may sustain that divergence by creating a barrier to recombination. We tested several lines of Escherichia coli, derived from a common ancestor and evolved for 20,000 generations, for their recombination ability. Some lines, but not others, had become mismatch repair-defective mutators during experimental evolution, providing different opportunities for DNA sequence divergence. We knocked out the repair system in lines that had retained this function, and we restored function to those lines that had become defective. We then estimated recombination rates in various crosses between these repair-deficient and -proficient strains. The effect of the mismatch repair system on recombination was greatest in those lines that had evolved nonfunctional repair, indicating they had undergone more sequence divergence and, consequently, were more sensitive to the recombination-inhibiting effect of a functional repair system. These results demonstrate the establishment of an incipient genetic barrier between formerly identical lines, and they support a model in which the mismatch repair system can influence speciation dynamics through its simultaneous effects on mutation and recombination. PMID- 10377418 TI - Widespread intra-serotype recombination in natural populations of dengue virus. AB - Diversity analysis of 71 published dengue virus gene sequences revealed several strains that appeared to be mosaics comprising gene regions with conflicting evolutionary histories. Subsequent maximum likelihood breakpoint estimation identified seven recombinants, including members of three of the four dengue virus serotypes, with breakpoints in the premembrane/membrane gene, the envelope gene, and at the junction of the envelope and first nonstructural genes. Many of the individual recombinants contain sequence representing separate genetic subtypes. The results were highly statistically significant and were confirmed by phylogenetic analysis of the regions of interest. These findings indicate that recombination may play a very significant role in shaping genetic diversity in dengue virus and, as such, have important implications for its biology and its control. PMID- 10377419 TI - Proposal for a standardized temporal scheme of biological classification for extant species. AB - With respect to conveying useful comparative information, current biological classifications are seriously flawed because they fail to (i) standardize criteria for taxonomic ranking and (ii) equilibrate assignments of taxonomic rank across disparate kinds of organisms. In principle, these problems could be rectified by adopting a universal taxonomic yardstick based on absolute dates of the nodes in evolutionary trees. By using procedures of temporal banding described herein, a simple philosophy of biological classification is proposed that would retain a manageable number of categorical ranks yet apply them in standardized fashion to time-dated phylogenies. The phylogenetic knowledge required for a time-standardized nomenclature arguably may emerge in the foreseeable future from vast increases in multilocus DNA sequence information (coupled with continued attention to phylogeny estimation from traditional systematic data). By someday encapsulating time-dated phylogenies in a familiar yet modified hierarchical ranking scheme, a temporal-banding approach would improve the comparative information content of biological classifications. PMID- 10377420 TI - Heterogeneous gene expression from the inactive X chromosome: an X-linked gene that escapes X inactivation in some human cell lines but is inactivated in others. AB - In mammalian females, most genes on one X chromosome are transcriptionally silenced as a result of X chromosome inactivation. Whereas it is well established that some X-linked genes "escape" X inactivation and are expressed from both active (Xa) and inactive (Xi) X chromosomes, most models for the chromosomal control of X-linked gene expression assume that the X inactivation status of a given gene is constant among different females within a population. In this report, we test the expression of human X-linked genes in primary cell lines from females with complete nonrandom X inactivation, by using transcribed polymorphisms to distinguish Xa and Xi expression. Six X-linked genes used to document this assay system showed monoallelic expression in all informative cell lines, consistent with X inactivation. However, a novel pattern of expression was observed for another gene, REP1; monoallelic expression, indicating inactivation, was detected in some lines, whereas biallelic expression, indicating escape from inactivation, was detected in others. Furthermore, levels of Xi expression varied among cell lines that expressed REP1. The cellular basis of Xi expression was examined by expression assays in single cells. These data indicate that REP1 is expressed from the Xi in all cells, but that the level of expression relative to Xa levels is reduced. These findings suggest that Xi gene expression is under a previously unsuspected level of genetic or epigenetic control, likely involving local or regional changes in chromatin organization that determine whether a gene escapes or is subject to X inactivation. PMID- 10377421 TI - Conditional requirement for the Flk-1 receptor in the in vitro generation of early hematopoietic cells. AB - Genetic studies in mice have previously demonstrated an intrinsic requirement for the vascular endothelial growth factor (VEGF) receptor Flk-1 in the early development of both the hematopoietic and endothelial cell lineages. In this study, embryonic stem (ES) cells homozygous for a targeted null mutation in flk-1 (flk-1 (-/-)) were examined for their hematopoietic potential in vitro during embryoid body (EB) formation or when cultured on the stromal cell line OP9. Surprisingly, in EB cultures flk-1 (-/-) ES cells were able to differentiate into all myeloid-erythroid lineages, albeit at half the frequency of heterozygous lines. In contrast, although flk-1 (-/-) ES cells formed mesodermal-like colonies on OP9 monolayers, they failed to generate hematopoietic clusters even in the presence of exogenous cytokines. However, flk-1 (-/-) OP9 cultures did contain myeloid precursors, albeit at greatly reduced percentages. This defect was rescued by first allowing flk-1 (-/-) ES cells to differentiate into EBs and then passaging these cells onto OP9 stroma. Thus, the requirement for Flk-1 in early hematopoietic development can be abrogated by alterations in the microenvironment. This finding is consistent with a role for Flk-1 in regulating the migration of early mesodermally derived precursors into a microenvironment that is permissive for hematopoiesis. PMID- 10377422 TI - Disruption of mRad50 causes embryonic stem cell lethality, abnormal embryonic development, and sensitivity to ionizing radiation. AB - The Mre11/Rad50 protein complex functions in diverse aspects of the cellular response to double-strand breaks (DSBs), including the detection of DNA damage, the activation of cell cycle checkpoints, and DSB repair. Whereas genetic analyses in Saccharomyces cerevisiae have provided insight regarding DSB repair functions of this highly conserved complex, the implication of the human complex in Nijmegen breakage syndrome reveals its role in cell cycle checkpoint functions. We established mRad50 mutant mice to examine the role of the mammalian Mre11/Rad50 protein complex in the DNA damage response. Early embryonic cells deficient in mRad50 are hypersensitive to ionizing radiation, consistent with a role for this complex in the repair of ionizing radiation-induced DSBs. However, the null mrad50 mutation is lethal in cultured embryonic stem cells and in early developing embryos, indicating that the mammalian Mre11/Rad50 protein complex mediates functions in normally growing cells that are essential for viability. PMID- 10377423 TI - Rapid p53 sequence analysis in primary lung cancer using an oligonucleotide probe array. AB - The p53 gene was sequenced in 100 primary human lung cancers by using direct dideoxynucleotide cycle sequencing and compared with sequence analysis by using the p53 GeneChip assay. Differences in sequence analysis between the two techniques were further evaluated to determine the accuracy and limitations of each method. p53 mutations were either detected by using both techniques or, if only detected by one technique, were confirmed by using mutation-specific oligonucleotide hybridization. Dideoxynucleotide sequencing of the conserved regions of the p53 gene (exons 5-9) detected 76% of the mutations within this region of the gene. The GeneChip p53 assay detected 81% of all (exons 2-11) mutations, including 80% of the mutations within the conserved regions of the gene. The GeneChip assay detected 46 of 52 missense mutations (88%), but 0 of 5 frameshift mutations. The specificity of direct sequencing and of the p53 GeneChip assay at detecting p53 mutations were 100% and 98%, respectively. The GeneChip p53 assay is a rapid and reasonably accurate approach for detecting p53 mutations; however, neither direct sequencing nor the p53 GeneChip are infallible at p53 mutation detection. PMID- 10377424 TI - Transglutaminase aggregates huntingtin into nonamyloidogenic polymers, and its enzymatic activity increases in Huntington's disease brain nuclei. AB - The protein huntingtin (htt), aggregated in neuronal nuclear inclusions, is pathognomonic of Huntington's disease (HD). Constructs, translated in vitro from the N terminus of htt, containing either polyQ23 from a normal individual, or polyQ41 or polyQ67 from an HD patient, were all soluble. Transglutaminase (TGase) crosslinked these proteins, and the aggregations did not have the staining properties of amyloid. More TGase-catalyzed aggregates formed when the polyglutamine domain of htt exceeded the pathologic threshold of polyQ36. Furthermore, shorter htt constructs, containing 135 aa or fewer, formed more aggregates than did larger htt constructs. TGase activity in the HD brain was increased compared with the control, with notable increases in cell nuclei. The increased TGase activity was brain specific. In lymphoblastoid cells from HD patients, TGase activity was decreased. TGase-mediated crosslinking of htt may be involved in the formation of the nonamyloidogenic nuclear inclusions found in the HD brain. The staining properties of nuclear inclusions in the HD brain revealed that they were not amyloid. PMID- 10377425 TI - Neurosecretory control of aging in Caenorhabditis elegans. AB - In the nematode Caenorhabditis elegans, an insulin receptor signaling pathway regulates adult life span and developmental arrest at the dauer larval stage. Here we show that the unc-64 and unc-31 genes also function in this pathway. These two genes are involved in mediating Ca2+-regulated secretion. Mutations in unc-64 and unc-31 increase adult life span and cause constitutive dauer formation. Both phenotypes are suppressed by mutations in daf-16, which also suppresses other mutations in this pathway. We present evidence that the site of action of unc-64 is neuronal, suggesting that a neurosecretory signal regulates life span and dauer formation. PMID- 10377426 TI - Stimulation of homologous recombination in plants by expression of the bacterial resolvase ruvC. AB - Targeted gene disruption exploits homologous recombination (HR) as a powerful reverse genetic tool, for example, in bacteria, yeast, and transgenic knockout mice, but it has not been applied to plants, owing to the low frequency of HR and the lack of recombinogenic mutants. To increase the frequency of HR in plants, we constructed transgenic tobacco lines carrying the Escherichia coli RuvC gene fused to a plant viral nuclear localization signal. We show that RuvC, encoding an endonuclease that binds to and resolves recombination intermediates (Holliday junctions) is properly transcribed in these lines and stimulates HR. We observed a 12-fold stimulation of somatic crossover between genomic sequences, a 11-fold stimulation of intrachromosomal recombination, and a 56-fold increase for the frequency of extrachromosomal recombination between plasmids cotransformed into young leaves via particle bombardment. This stimulating effect may be transferred to any plant species to obtain recombinogenic plants and thus constitutes an important step toward gene targeting. PMID- 10377428 TI - Colinearity and its exceptions in orthologous adh regions of maize and sorghum. AB - Orthologous adh regions of the sorghum and maize genomes were sequenced and analyzed. Nine known or candidate genes, including adh1, were found in a 225 kilobase (kb) maize sequence. In a 78-kb space of sorghum, the nine homologues of the maize genes were identified in a colinear order, plus five additional genes. The major fraction of DNA in maize, occupying 166 kb (74%), is represented by 22 long terminal repeat (LTR) retrotransposons. About 6% of the sequence belongs to 33 miniature inverted-repeat transposable elements (MITEs), remnants of DNA transposons, 4 simple sequence repeats, and low-copy-number DNAs of unknown origin. In contrast, no LTR retroelements were detected in the orthologous sorghum region. The unconserved sorghum DNA is composed of 20 putative MITEs, transposon-like elements, 5 simple sequence repeats, and low-copy-number DNAs of unknown origin. No MITEs were discovered in the 166 kb of DNA occupied by the maize LTR retrotransposons. In both species, MITEs were found in the space between genes and inside introns, indicating specific insertion and/or retention for these elements. Two adjacent sorghum genes, including one gene missing in maize, had colinear homologues on Arabidopsis chromosome IV, suggesting two rearrangements in the sorghum and three in the maize genome in comparison to a four-gene region of Arabidopsis. Hence, multiple small rearrangements may be present even in largely colinear genomic regions. These studies revealed a much higher degree of diversity at a microstructural level than predicted by genetic mapping studies for closely related grass species, as well as for comparisons of monocots and dicots. PMID- 10377427 TI - The natriuretic peptide clearance receptor locally modulates the physiological effects of the natriuretic peptide system. AB - Natriuretic peptides (NPs), mainly produced in heart [atrial (ANP) and B-type (BNP)], brain (CNP), and kidney (urodilatin), decrease blood pressure and increase salt excretion. These functions are mediated by natriuretic peptide receptors A and B (NPRA and NPRB) having cytoplasmic guanylyl cyclase domains that are stimulated when the receptors bind ligand. A more abundantly expressed receptor (NPRC or C-type) has a short cytoplasmic domain without guanylyl cyclase activity. NPRC is thought to act as a clearance receptor, although it may have additional functions. To test how NPRC affects the cardiovascular and renal systems, we inactivated its gene (Npr3) in mice by homologous recombination. The half life of [125I]ANP in the circulation of homozygotes lacking NPRC is two thirds longer than in the wild type, although plasma levels of ANP and BNP in heterozygotes and homozygotes are close to the wild type. Heterozygotes and homozygotes have a progressively reduced ability to concentrate urine, exhibit mild diuresis, and tend to be blood volume depleted. Blood pressure in the homozygotes is 8 mmHg (1 mmHg = 133 Pa) below normal. These results are consistent with the sole cardiovascular/renal function of NPRC being to clear natriuretic peptides, thereby modulating local effects of the natriuretic peptide system. Unexpectedly, Npr3 -/- homozygotes have skeletal deformities associated with a considerable increase in bone turnover. The phenotype is consistent with the bone function of NPRC being to clear locally synthesized CNP and modulate its effects. We conclude that NPRC modulates the availability of the natriuretic peptides at their target organs, thereby allowing the activity of the natriuretic peptide system to be tailored to specific local needs. PMID- 10377429 TI - Efficient export of the glucose transporter Hxt1p from the endoplasmic reticulum requires Gsf2p. AB - Mutations in the GSF2 gene cause glucose starvation phenotypes in Saccharomyces cerevisiae. We have isolated the HXT1 gene, which encodes a low-affinity, high capacity glucose transporter, as a multicopy suppressor of a gsf2 mutation. We show that gsf2 mutants accumulate Hxt1p in the endoplasmic reticulum (ER) and that Gsf2p is a 46-kDa integral membrane protein localized to the ER. gsf2 mutants also display a galactose growth defect and abnormal localization of the galactose transporter Gal2p but are not defective in function or localization of the high-affinity glucose transporter Hxt2p. These findings suggest that Gsf2p functions in the ER to promote the secretion of certain hexose transporters. PMID- 10377430 TI - Protein kinase B/Akt-mediated phosphorylation promotes nuclear exclusion of the winged helix transcription factor FKHR1. AB - Although genetic analysis has demonstrated that members of the winged helix, or forkhead, family of transcription factors play pivotal roles in the regulation of cellular differentiation and proliferation, both during development and in the adult, little is known of the mechanisms underlying their regulation. Here we show that the activation of phosphatidylinositol 3 (PI3) kinase by extracellular growth factors induces phosphorylation, nuclear export, and transcriptional inactivation of FKHR1, a member of the FKHR subclass of the forkhead family of transcription factors. Protein kinase B (PKB)/Akt, a key mediator of PI3 kinase signal transduction, phosphorylated recombinant FKHR1 in vitro at threonine-24 and serine-253. Mutants FKHR1(T24A), FKHR1(S253A), and FKHR1(T24A/S253A) were resistant to both PKB/Akt-mediated phosphorylation and PI3 kinase-stimulated nuclear export. These results indicate that phosphorylation by PKB/Akt negatively regulates FKHR1 by promoting export from the nucleus. PMID- 10377431 TI - The PTEN tumor suppressor homolog in Caenorhabditis elegans regulates longevity and dauer formation in an insulin receptor-like signaling pathway. AB - Inactivation of the tumor suppressor PTEN gene is found in a variety of human cancers and in cancer predisposition syndromes. Recently, PTEN protein has been shown to possess phosphatase activity on phosphatidylinositol 3,4,5 trisphosphate, a product of phosphatidylinositol 3-kinase. We have identified a homolog of PTEN in Caenorhabditis elegans and have found that it corresponds to the daf-18 gene, which had been defined by a single, phenotypically weak allele, daf-18(e1375). By analyzing an allele, daf-18(nr2037), which bears a deletion of the catalytic portion of CePTEN/DAF-18, we have shown that mutation in daf-18 can completely suppress the dauer-constitutive phenotype caused by inactivation of daf-2 or age-1, which encode an insulin receptor-like molecule and the catalytic subunit of phosphatidylinositol 3-kinase, respectively. In addition, daf 18(nr2037) dramatically shortens lifespan, both in a wild-type background and in a daf-2 mutant background that normally prolongs lifespan. The lifespan in a daf 18(nr2037) mutant can be restored to essentially that of wild type when combined with a daf-2 mutation. Our studies provide genetic evidence that, in C. elegans, the PTEN homolog DAF-18 functions as a negative regulator of the DAF-2 and AGE-1 signaling pathway, consistent with the notion that DAF-18 acts a phosphatidylinositol 3,4,5-trisphosphate phosphatase in vivo. Furthermore, our studies have uncovered a longevity-promoting activity of the PTEN homolog in C. elegans. PMID- 10377432 TI - An engineered closterovirus RNA replicon and analysis of heterologous terminal sequences for replication. AB - Citrus tristeza virus (CTV) populations in citrus trees are unusually complex mixtures of viral genotypes and defective RNAs developed during the long-term vegetative propagation of the virus and by additional mixing by aphid transmission. The viral replication process allows the maintenance of minor amounts of disparate genotypes and defective RNAs in these populations. CTV is a member of the Closteroviridae possessing a positive-stranded RNA genome of approximately 20 kilobases that expresses the replicase-associated genes as an approximately 400-kDa polyprotein and the remaining 10 3' genes through subgenomic mRNAs. A full-length cDNA clone of CTV was generated from which RNA transcripts capable of replication in protoplasts were derived. The large size of cDNA hampered its use as a genetic system. Deletion of 10 3' genes resulted in an efficient RNA replicon that was easy to manipulate. To investigate the origin and maintenance of the genotypes in CTV populations, we tested the CTV replicase for its acceptance of divergent sequences by creating chimeric replicons with heterologous termini and examining their ability to replicate. Exchange of the similar 3' termini resulted in efficient replication whereas substitution of the divergent (up to 58% difference in sequence) 5' termini resulted in reduced but significant replication, generally in proportion to the extent of sequence divergence. PMID- 10377434 TI - Conformational variants of class II MHC/peptide complexes induced by N- and C terminal extensions of minimal peptide epitopes. AB - Class II MHC molecules are known to exist in conformational variants. "Floppy" and "compact" forms of murine MHC molecules, for example, are discriminated by their migration behavior on SDS/PAGE and represent empty and ligand-loaded forms. Here we show that formation of distinctly faster-migrating ligand complexes (F forms) rather than the normal compact (C-) forms of HLA-DR1 or -DR4 results from extensions of minimal peptide epitopes (such as HA306-318 or IC106-120) by approximately 10 amino acids at either the N or the C terminus. Two similar but distinct F-forms (FI and FII) were detected, depending on the site of the extension. Both F-forms were characterized by increased surface hydrophobicity and reduced SDS-stability. Native gel separations and size exclusion chromatography indicated that the F-forms had increased hydrodynamic radii compared with the C-form and an apparent size similar to that of empty MHC molecules. The regions on the ligand overhangs responsible for the effect began at a distance of approximately 5 amino acids on either side of the epitopes, comprised 4-8 amino acids (i.e., a total overhang of 9-14), and did not have a particular sequence preference. The possible functional significance of these forms is discussed. PMID- 10377433 TI - Induction of differentiation of pre-NKT cells to mature Valpha14 NKT cells by granulocyte/macrophage colony-stimulating factor. AB - Valpha14 NKT cells express an invariant antigen receptor encoded by Valpha14 and Jalpha281 gene segments as well as natural killer (NK) markers, including NK1.1. Here, we describe a precursor population of NKT cells (pre-NKT) that expresses NK1.1, T cell antigen receptor beta, pTalpha, and RAG1/2 but not Valpha14 and surface CD3epsilon. Such pre-NKT cells were differentiated successfully in vitro into mature CD3epsilon+ Valpha14(+) NKT cells by IL-15 and granulocyte/macrophage colony-stimulating factor (GM-CSF) in conjunction with stroma cells. Interestingly, only GM-CSF without stroma cells induced the Valpha14-Jalpha281 gene rearrangement in the pre-NKT cells. This also was confirmed by the findings that the number of mature Valpha14 NKT cells and the frequency of Valpha14 Jalpha281 rearrangements were decreased significantly in the mice lacking a GM CSF receptor component, common beta-chain. These results suggest a crucial role of GM-CSF in the development of Valpha14 NKT cells in vivo. PMID- 10377435 TI - Cytolytic and IFN-gamma-producing activities of gamma delta T cells in the mouse intestinal epithelium are T cell receptor-beta-chain dependent. AB - We analyzed the cytolytic activity of intraepithelial T cells (IEL) isolated from the small intestines of 2- to 3-month-old mutant mice rendered deficient in different gene(s) in which the number of IEL expressing either T cell receptor (TCR)-alpha beta (alpha beta-IEL) or TCR-gamma delta (gamma delta-IEL) were absent or markedly diminished. When compared with wild-type littermates, cytolytic activity of gamma delta-IEL was sharply attenuated in TCR-beta mutant mice but remained unaltered in TCR-alpha mutant mice in which a minor population of dull TCR-beta+ (betadim)-IEL was also present. Cytolytic activity of gamma delta-IEL was maintained in mice doubly homozygous for beta2-microglobulin and transporter associated with antigen processing 1 gene mutations in which a conspicuous decrease was noted in absolute numbers of alpha beta-IEL. In contrast, both TCR-delta and IL-7 receptor-alpha gene mutations that lead to lack of gamma delta-IEL generation did not affect the development or cytolytic activity of the remaining alpha beta-IEL. The anti-CD3 and anti-TCR-gamma delta mAb-induced IFN-gamma production of gamma delta-IEL showed the same TCR-alpha and TCR-beta mutation-dependent variability. These results indicate that cytolytic and IFN-gamma-producing activities of gamma delta T cells in mouse intestinal epithelium are TCR-beta-chain-dependent. PMID- 10377436 TI - Cancer-specific chromosome alterations in the constitutive fragile region FRA3B. AB - We have sequenced 870 kilobases of the FHIT/FRA3B locus, from FHIT intron 3 to intron 7. The locus is AT rich (61.5%) and Alu poor (6. 2%), and it apparently does not harbor other genes. In a detailed analysis of the 308-kilobase region between FHIT exon 5 and the telomeric end of intron 3, a region known to encompass a human papillomavirus-16 integration site and two clusters of aphidicolin-induced chromosome 3p14.2 breakpoints, we have precisely mapped 10 deletion and translocation endpoints in cancer-derived cell lines relative to positions of specific repetitive elements, regions of high genome flexibility and aphidicolin-induced breakpoints. Conclusions are (i) that aphidicolin-induced breakpoint clusters fall close to high-flexibility sequences, suggesting that these sequences contribute directly to aphidicolin-induced fragility; (ii) that 9 of the 10 FHIT allelic deletions in cancer cell lines resulted in loss of exons, with 7 deletion endpoints near long interspersed nuclear elements or long terminal repeat elements; and (iii) that cancer-specific deletions encompass multiple high-flexibility genomic regions, suggesting that fragile breaks may occur at these regions, whereas repair of the breaks involves homologous pairing of flanking sequences with concomitant deletion of the damaged fragile sequence. PMID- 10377437 TI - Treatment of noninfectious intermediate and posterior uveitis with the humanized anti-Tac mAb: a phase I/II clinical trial. AB - To evaluate the safety and potential therapeutic activity of humanized anti-IL-2 receptor mAb (Daclizumab) therapy in the treatment of patients with severe, sight threatening, intermediate and posterior noninfectious uveitis, a nonrandomized, open-label, pilot study was performed. Patients with uveitis were treated with a minimum of 20 mg of prednisone, cyclosporine, antimetabolites, or any combination of these agents were eligible. Patients were weaned off their systemic immunosuppressive agents according to a standardized schedule, while ultimately receiving Daclizumab infusions every 4 weeks. Anti-IL-2 receptor antibody therapy, given intravenously with intervals of up to 4 weeks in lieu of standard immunosuppressive therapy, appeared to prevent the expression of severe sight threatening intraocular inflammatory disease in 8 of 10 patients treated over a 12-month period, with noted improvements in visual acuity. One patient met a primary endpoint with a loss of vision of 10 letters or more from baseline in one eye and another patient discontinued therapy because of evidence of increased ocular inflammation. All patients were able to tolerate the study medications without the need for dose reduction. We report effective long-term use of anti-IL 2 therapy for an autoimmune indication. These initial findings would suggest that anti-IL-2 receptor therapy may be an effective therapeutic approach for uveitis and, by implication, other disorders with a predominant Th1 profile. PMID- 10377438 TI - Modulation of TEL transcription activity by interaction with the ubiquitin conjugating enzyme UBC9. AB - The E-26 transforming specific (ETS)-related gene TEL, also known as ETV6, is involved in a large number of chromosomal rearrangements associated with leukemia and congenital fibrosarcoma. The encoded protein contains two functional domains: a helix-loop-helix (HLH) domain (also known as pointed domain) located at the N terminus and a DNA-binding domain located at the C terminus. The HLH domain is involved in protein-protein interaction with itself and other members of the ETS family of transcription factors such as FLI1. TEL is a transcription factor, and we and others have shown that it is a repressor of gene expression. To understand further the role of TEL in the cell, we have used an in vivo interaction system to identify proteins that interact with TEL. We show that a protein, UBC9, interacts specifically with TEL in vitro and in vivo. UBC9 is a member of the family of ubiquitin-conjugating enzymes. These enzymes usually are involved in proteosome-mediated degradation; however, our data suggest that interaction of TEL with UBC9 does not lead to TEL degradation. Our studies show that UBC9 binds to TEL exclusively through the HLH domain of TEL. We also show that TEL expressed as fusion to the DNA-binding domain of Gal4 completely represses a Gal4 responsive promoter, but that the coexpression of UBC9 in the same system restores the activity of the promoter. Targeted point mutation of conserved amino acids in UBC9 essential for enzymatic ubiquitination of proteins does not affect interaction nor transcriptional activity. Based on our data, we conclude that UBC9 physically interacts with TEL through the HLH domain and that the interaction leads to modulation of the transcription activity of TEL. PMID- 10377439 TI - A critical role for the peroxisome proliferator-activated receptor alpha (PPARalpha) in the cellular fasting response: the PPARalpha-null mouse as a model of fatty acid oxidation disorders. AB - We hypothesized that the lipid-activated transcription factor, the peroxisome proliferator-activated receptor alpha (PPARalpha), plays a pivotal role in the cellular metabolic response to fasting. Short-term starvation caused hepatic steatosis, myocardial lipid accumulation, and hypoglycemia, with an inadequate ketogenic response in adult mice lacking PPARalpha (PPARalpha-/-), a phenotype that bears remarkable similarity to that of humans with genetic defects in mitochondrial fatty acid oxidation enzymes. In PPARalpha+/+ mice, fasting induced the hepatic and cardiac expression of PPARalpha target genes encoding key mitochondrial (medium-chain acyl-CoA dehydrogenase, carnitine palmitoyltransferase I) and extramitochondrial (acyl-CoA oxidase, cytochrome P450 4A3) enzymes. In striking contrast, the hepatic and cardiac expression of most PPARalpha target genes was not induced by fasting in PPARalpha-/- mice. These results define a critical role for PPARalpha in a transcriptional regulatory response to fasting and identify the PPARalpha-/- mouse as a potentially useful murine model of inborn and acquired abnormalities of human fatty acid utilization. PMID- 10377440 TI - X inactivation and somatic cell selection rescue female mice carrying a Piga-null mutation. AB - A somatic mutation in the X linked PIGA gene is responsible for the deficiency of glycosyl phosphatidylinositol (GPI)-anchored proteins on blood cells from patients with paroxysmal nocturnal hemoglobinuria. No inherited form of GPI anchor deficiency has been described. Because conventional Piga gene knockout is associated with high embryonic lethality in chimeric mice, we used the Cre/loxP system. We generated mice in which two loxP sites flank part of Piga exon 2. After crossbreeding with female mice of the EIIa-cre strain, the floxed allele undergoes Cre-mediated recombination with high efficiency during early embryonic development. Because of X chromosome inactivation, female offspring are mosaic for cells that express or lack GPI-linked proteins. Analysis of mosaic mice showed that in heart, lung, kidney, brain, and liver, mainly wild-type Piga is active, suggesting that these tissues require GPI-linked proteins. The salient exceptions were spleen, thymus, and red blood cells, which had almost equal numbers of cells expressing the wild-type or the recombined allele, implying that GPI-linked proteins are not essential for the derivation of these tissues. PIGA( ) cells had no growth advantage, suggesting that other factors are needed for their clonal dominance in patients with paroxysmal nocturnal hemoglobinuria. PMID- 10377441 TI - JC virus DNA is present in the mucosa of the human colon and in colorectal cancers. AB - JC virus (JCV) is a polyoma virus that commonly infects humans. We have found T antigen DNA sequences of JCV in the mucosa of normal human colons, colorectal cancers, colorectal cancer xenografts raised in nude mice, and in the human colon cancer cell line SW480. A larger number of viral copies is present in cancer cells than in non-neoplastic colon cells, and sequence microheterogeneity occurs within individual colonic mucosal specimens. The improved yield of detection after treatment with topoisomerase I suggests that the viral DNA is negatively supercoiled in the human tissues. These results indicate that JCV DNA can be found in colonic tissues, which raises the possibility that this virus may play a role in the chromosomal instability observed in colorectal carcinogenesis. PMID- 10377442 TI - Silibinin decreases prostate-specific antigen with cell growth inhibition via G1 arrest, leading to differentiation of prostate carcinoma cells: implications for prostate cancer intervention. AB - Reduction in serum prostate-specific antigen (PSA) levels has been proposed as an endpoint biomarker for hormone-refractory human prostate cancer intervention. We examined whether a flavonoid antioxidant silibinin (an active constituent of milk thistle) decreases PSA levels in hormone-refractory human prostate carcinoma LNCaP cells and whether this effect has biological relevance. Silibinin treatment of cells grown in serum resulted in a significant decrease in both intracellular and secreted forms of PSA concomitant with a highly significant to complete inhibition of cell growth via a G1 arrest in cell cycle progression. Treatment of cells grown in charcoal-stripped serum and 5alpha-dihydrotestosterone showed that the observed effects of silibinin are those involving androgen-stimulated PSA expression and cell growth. Silibinin-induced G1 arrest was associated with a marked decrease in the kinase activity of cyclin-dependent kinases (CDKs) and associated cyclins because of a highly significant decrease in cyclin D1, CDK4, and CDK6 levels and an induction of Cip1/p21 and Kip1/p27 followed by their increased binding with CDK2. Silibinin treatment of cells did not result in apoptosis and changes in p53 and bcl2, suggesting that the observed increase in Cip1/p21 is a p53-independent effect that does not lead to an apoptotic cell death pathway. Conversely, silibinin treatment resulted in a significant neuroendocrine differentiation of LNCaP cells as an alternative pathway after Cip1/p21 induction and G1 arrest. Together, these results suggest that silibinin could be a useful agent for the intervention of hormone-refractory human prostate cancer. PMID- 10377443 TI - Constitutive cell surface association between CD4 and CCR5. AB - HIV-1 entry into cells involves formation of a complex between gp120 of the viral envelope glycoprotein (Env), a receptor (CD4), and a coreceptor. For most strains of HIV, this coreceptor is CCR5. Here, we provide evidence that CD4 is specifically associated with CCR5 in the absence of gp120 or any other receptor specific ligand. The amount of CD4 coimmunoprecipitated with CCR5 was significantly higher than that with the other major HIV coreceptor, CXCR4, and in contrast to CXCR4 the CD4-CCR5 coimmunoprecipitation was not significantly increased by gp120. The CD4-CCR5 interaction probably takes place via the second extracellular loop of CCR5 and the first two domains of CD4. It can be inhibited by CCR5- and CD4-specific antibodies that interfere with HIV-1 infection, indicating a possible role in virus entry. These findings suggest a possible pathway of HIV-1 evolution and development of immunopathogenicity, a potential new target for antiretroviral drugs and a tool for development of vaccines based on Env-CD4-CCR5 complexes. The constitutive association of a seven-transmembrane domain G protein-coupled receptor with another receptor also indicates new possibilities for cross-talk between cell surface receptors. PMID- 10377445 TI - Murine gammaherpesvirus M2 gene is latency-associated and its protein a target for CD8(+) T lymphocytes. AB - Murine gammaherpesvirus 68 (MHV-68) infection of mice is a potential model with which to address fundamental aspects of the pathobiology and host control of gammaherpesvirus latency. Control of MHV-68 infection, like that of Epstein-Barr virus, is strongly dependent on the cellular immune system. However, the molecular biology of MHV-68 latency is largely undefined. A screen of the MHV-68 genome for potential latency-associated mRNAs revealed that the region encompassing and flanking the genomic terminal repeats is transcriptionally active in the latently infected murine B-cell tumor line S11. Transcription of one MHV-68 gene, that encoding the hypothetical M2 protein, was detected in virtually all latently infected S11 cells and in splenocytes of latently infected mice, but not in lytically infected fibroblasts. Furthermore, an epitope was identified in the predicted M2 protein that is recognized by CD8(+) T cells from infected mice and a cytotoxic T lymphocyte line that recognizes this epitope killed S11 cells, indicating that the M2 protein is expressed during latent infection and is a target for the host cytotoxic T lymphocyte response. This work therefore provides essential information for modeling MHV-68 latency and strategies of immunotherapy against gammaherpesvirus-related diseases in a highly tractable animal model. PMID- 10377444 TI - Virulent Salmonella typhimurium has two periplasmic Cu, Zn-superoxide dismutases. AB - Periplasmic Cu, Zn-cofactored superoxide dismutase (SodC) protects Gram-negative bacteria from exogenous oxidative damage. The virulent Salmonella typhimurium strain ATCC 14028s has been found to contain two discrete periplasmic Cu, Zn-SOD enzymes that are only 57% identical at the amino acid level. SodCI is carried by a cryptic bacteriophage, and SodCII is closely related to the Cu, Zn-superoxide dismutase of Escherichia coli. All Salmonella serotypes appear to carry the sodCII locus, but the phage-associated sodCI gene is found only in certain strains belonging to the most highly pathogenic serotypes. Expression of either sodC locus appears to be enhanced during stationary phase, but only sodCII is regulated by the alternative sigma factor sigmas (RpoS). Mutants lacking both sodC genes are less lethal for mice than mutants possessing either sodC locus alone, indicating that both Cu, Zn-SOD enzymes contribute to Salmonella pathogenicity. The evolutionary acquisition of an additional sodC gene has contributed to the enhanced virulence of selected Salmonella strains. PMID- 10377446 TI - The neural substrate and temporal dynamics of interference effects in working memory as revealed by event-related functional MRI. AB - Research on the prefrontal cortex (PFC) of monkeys and humans indicates that this region supports a heterogeneous repertoire of mental processes that contribute to many complex behaviors, such as working memory. Anatomical evidence for some of these processes derives from functional neuroimaging experiments using blocked experimental designs, which average signal across all components of many trials and therefore cannot dissociate distinct processes with different time courses. Using event-related functional MRI, we were able to isolate temporally the neural correlates of processes contributing to the target presentation, delay, and probe portions of an item-recognition task. Two types of trials were of greatest interest: those with Recent Negative probes that matched an item from the target set of the previous, but not the present, two trials, and those with Nonrecent Negative probes that did not match a target item from either the present or the two previous trials. There was no difference between the two trial types in target presentation (i.e., encoding) or delay-period (i.e., active maintenance) PFC activation, but there was significantly greater activation for Recent Negatives than Nonrecent Negative activation associated with the probe period within left ventrolateral PFC. These findings characterize spatially and temporally a proactive interference effect that may reflect the operation of a PFC-mediated response-inhibition mechanism that contributes to working memory performance. PMID- 10377447 TI - Relationship between presynaptic calcium transients and postsynaptic currents at single gamma-aminobutyric acid (GABA)ergic boutons. AB - Postsynaptic responses to stereotyped activation of single axons are known to fluctuate, but the origin of synaptic variability in the vertebrate central nervous system is still unclear. To test the hypothesis that fluctuations of inhibitory postsynaptic currents reflect variations in presynaptic Ca2+ concentration, we examined single GABAergic axodendritic contacts in low-density cultures. Collicular neurons from rat embryos were loaded with the Ca2+ indicator Oregon Green 488 BAPTA-1. Presynaptic axon terminals were visualized by staining with the styryl dye RH414. Under the condition of action potential block, RH414 labeled boutons were activated selectively by current pulses applied through a fine-tipped glass pipette. Short (1- to 3-ms) depolarization of isolated boutons resulted in stimulus-locked changes of presynaptic Ca2+ concentration ([Ca2+]pre) and in evoked inhibitory postsynaptic currents (eIPSCs). Varying the strength of the stimulating currents produced a wide amplitude range of both presynaptic fluorescence transients (up to 220% of the resting value) and postsynaptic conductance changes (up to 2-3 nS). It was found that average eIPSCs displayed an approximately third-power dependency on [Ca2+]pre. Transmitter release retained its probabilistic character throughout the range of observed [Ca2+]pre values. In any tested single bouton, maximal eIPSCs occurred in association with the largest [Ca2+]pre transients, but failures were present at any [Ca2+]pre. The increase of maximal eIPSC amplitudes in connection with higher [Ca2+]pre supports the hypothesis that GABAergic boutons have the capacity to regulate synaptic strength by changing the number of simultaneously released vesicles. PMID- 10377448 TI - Interaction between astrocytes and adult subventricular zone precursors stimulates neurogenesis. AB - Neurogenesis continues in the mammalian subventricular zone (SVZ) throughout life. However, the signaling and cell-cell interactions required for adult SVZ neurogenesis are not known. In vivo, migratory neuroblasts (type A cells) and putative precursors (type C cells) are in intimate contact with astrocytes (type B cells). Type B cells also contact each other. We reconstituted SVZ cell-cell interactions in a culture system free of serum or exogenous growth factors. Culturing dissociated postnatal or adult SVZ cells on astrocyte monolayers-but not other substrates-supported extensive neurogenesis similar to that observed in vivo. SVZ precursors proliferated rapidly on astrocytes to form colonies containing up to 100 type A neuroblasts. By fractionating the SVZ cell dissociates with differential adhesion to immobilized polylysine, we show that neuronal colony-forming precursors were concentrated in a fraction enriched for type B and C cells. Pure type A cells could migrate in chains but did not give rise to neuronal colonies. Because astrocyte-conditioned medium alone was not sufficient to support SVZ neurogenesis, direct cell-cell contact between astrocytes and SVZ neuronal precursors may be necessary for the production of type A cells. PMID- 10377449 TI - Mice lacking complex gangliosides develop Wallerian degeneration and myelination defects. AB - Gangliosides are a family of sialic acid-containing glycosphingolipids highly enriched in the mammalian nervous system. Although they are the major sialoglycoconjugates in the brain, their neurobiological functions remain poorly defined. By disrupting the gene for a key enzyme in complex ganglioside biosynthesis (GM2/GD2 synthase; EC 2.4.1.92) we generated mice that express only simple gangliosides (GM3/GD3) and examined their central and peripheral nervous systems. The complex ganglioside knockout mice display decreased central myelination, axonal degeneration in both the central and peripheral nervous systems, and demyelination in peripheral nerves. The pathological features of their nervous system closely resemble those reported in mice with a disrupted gene for myelin-associated glycoprotein (MAG), a myelin receptor that binds to complex brain gangliosides in vitro. Furthermore, GM2/GD2 synthase knockout mice have reduced MAG expression in the central nervous system. These results indicate that complex gangliosides function in central myelination and maintaining the integrity of axons and myelin. They also support the theory that complex gangliosides are endogenous ligands for MAG. The data extend and clarify prior observations on a similar mouse model, which reported only subtle conduction defects in their nervous system [Takamiya, K., Yamamoto, A., Furukawa, K., Yamashiro, S., Shin, M., Okada, M., Fukumoto, S., Haraguchi, M., Takeda, N., Fujimura, K., et al. (1996) Proc. Natl. Acad. Sci. USA 93, 10662-10667]. PMID- 10377450 TI - A brain sexual dimorphism controlled by adult circulating androgens. AB - Reports of structural differences between the brains of men and women, heterosexual and homosexual men, and male-to-female transsexuals and other men have been offered as evidence that the behavioral differences between these groups are likely caused by differences in the early development of the brain. However, a possible confounding variable is the concentration of circulating hormones seen in these groups in adulthood. Evaluation of this possibility hinges on the extent to which circulating hormones can alter the size of mammalian brain regions as revealed by Nissl stains. We now report a sexual dimorphism in the volume of a brain nucleus in rats that can be completely accounted for by adult sex differences in circulating androgen. The posterodorsal nucleus of the medial amygdala (MePD) has a greater volume in male rats than in females, but adult castration of males causes the volume to shrink to female values within four weeks, whereas androgen treatment of adult females for that period enlarges the MePD to levels equivalent to normal males. This report demonstrates that adult hormone manipulations can completely reverse a sexual dimorphism in brain regional volume in a mammalian species. The sex difference and androgen responsiveness of MePD volume is reflected in the soma size of neurons there. PMID- 10377451 TI - Gap-junctional coupling between neurons and astrocytes in primary central nervous system cultures. AB - Gap-junctional communication between neurons and astrocytes dissociated from rat brain was identified in culture by using dye-transfer assays and electrophysiological measurements. Cell types were identified by using antibodies against beta-tubulin III, glial fibrillary acidic protein, and 2',3'-cyclic nucleotide phosphohydrolase, which are antigenic determinants of neurons, astroglia, and oligodendrocytes, respectively. Dye coupling was examined as a function of time after dissociated embryonic brain cells were plated onto confluent monolayers of postnatal astrocytes by intracellularly injecting the fluorochrome Lucifer yellow. Coupling of neurons to the astrocytic monolayer was most frequent between 48 h and 72 h in culture and declined over the next 4 days. This gradual uncoupling was accompanied by progressive neuronal maturation, as indicated by morphological measurements in camera lucida drawings. Dye spread was abolished reversibly by octanol, an agent that blocks gap junction channels in other systems. Double whole-cell voltage-clamp measurements confirmed the presence of heterocellular electrical coupling in these cocultures. Coupling was also seen between neurons and astrocytes in cocultures of cells dissociated from embryonic cerebral hemispheres but was rarely detectable in cocultures of postnatal brain cells. These data strongly suggest that junctional communication may provide metabolic and electrotonic interconnections between neuronal and astrocytic networks at early stages of neural development and that such interactions are weakened as differentiation progresses. PMID- 10377452 TI - Wild-type but not Alzheimer-mutant amyloid precursor protein confers resistance against p53-mediated apoptosis. AB - Amyloid precursor proteins (APPs) are expressed in multiple organs and cell types in diverse species. Their conservation across species and high abundance in brain and the association of various APP missense mutations with autosomal dominant forms of familial Alzheimer's disease (FAD) suggest important roles for APP in the central nervous system. However, the basic functions of APP in the central nervous system remain largely unknown. To assess potential effects of APP on neuronal death and survival, we transfected APP-deficient rat neuroblastoma cells (B103) with DNA constructs encoding wild-type or FAD-mutant human APP. Wild-type, but not FAD-mutant, APP effectively protected cells against apoptosis induced by ultraviolet irradiation, staurosporine, or p53. Wild-type APP also strongly inhibited p53 DNA-binding activity and p53-mediated gene transactivation, whereas FAD-mutant APP did not. We conclude that APP protects neuronal cells against apoptosis by controlling p53 activation at the post-translational level. Disruption of this function by mutations or alterations in APP processing could enhance neuronal vulnerability to secondary insults and contribute to neuronal degeneration. PMID- 10377453 TI - Selective loss of cone function in mice lacking the cyclic nucleotide-gated channel CNG3. AB - Two types of photoreceptors, rods and cones, coexist in the vertebrate retina. An in-depth analysis of the retinal circuitry that transmits rod and cone signals has been hampered by the presence of intimate physical and functional connections between rod and cone pathways. By deleting the cyclic nucleotide-gated channel CNG3 we have generated a mouse lacking any cone-mediated photoresponse. In contrast, the rod pathway is completely intact in CNG3-deficient mice. The functional loss of cone function correlates with a progressive degeneration of cone photoreceptors but not of other retinal cell types. CNG3-deficient mice provide an animal model to dissect unequivocally the contribution of rod and cone pathways for normal retinal function. PMID- 10377454 TI - Somatotopy of the lateral line projection in larval zebrafish. AB - We examined the topography of the lateral line primary projection in zebrafish larvae by double labeling. The projections of two identified neuromasts of the posterior lateral line are seen as two separate sets of fibers that show reproducible spatial relationships: the projection of the anterior neuromast is always ventrolateral to that of a more posteriorly located neuromast. The same rule applies to the projection of anterior lateral line neuromasts. The position of the neuromasts along the antero posterior axis of the fish therefore is represented in the central projection of the sensory neurons. This somatotopy is similar to, and may be at the origin of, the tonotopic projection of the cochlear hair cells in mammals. PMID- 10377456 TI - Activation of erythropoietin receptor in the absence of hormone by a peptide that binds to a domain different from the hormone binding site. AB - Applying a homology search method previously described, we identified a sequence in the extracellular dimerization site of the erythropoietin receptor, distant from the hormone binding site. A peptide identical to that sequence was synthesized. Remarkably, it activated receptor signaling in the absence of erythropoietin. Neither the peptide nor the hormone altered the affinity of the other for the receptor; thus, the peptide does not bind to the hormone binding site. The combined activation of signal transduction by hormone and peptide was strongly synergistic. In mice, the peptide acted like the hormone, protecting against the decrease in hematocrit caused by carboplatin. PMID- 10377455 TI - Nonsteroid drug selectivities for cyclo-oxygenase-1 rather than cyclo-oxygenase-2 are associated with human gastrointestinal toxicity: a full in vitro analysis. AB - The beneficial actions of nonsteroid anti-inflammatory drugs (NSAID) can be associated with inhibition of cyclo-oxygenase (COX)-2 whereas their harmful side effects are associated with inhibition of COX-1. Here we report data from two related assay systems, the human whole blood assay and a modified human whole blood assay (using human A549 cells as a source of COX-2). This assay we refer to as the William Harvey Modified Assay. Our aim was to make meaningful comparisons of both classical NSAIDs and newer COX-2-selective compounds. These comparisons of the actions of >40 NSAIDs and novel COX-2-selective agents, including celecoxib, rofecoxib and diisopropyl fluorophosphate, demonstrate a distribution of compound selectivities toward COX-1 that aligns with the risk of serious gastrointestinal complications. In conclusion, this full in vitro analysis of COX 1/2 selectivities in human tissues clearly supports the theory that inhibition of COX-1 underlies the gastrointestinal toxicity of NSAIDs in man. PMID- 10377457 TI - Suppression of the heterotrimeric G protein causes abnormal morphology, including dwarfism, in rice. AB - Transgenic rice containing an antisense cDNA for the alpha subunit of rice heterotrimeric G protein produced little or no mRNA for the subunit and exhibited abnormal morphology, including dwarf traits and the setting of small seeds. In normal rice, the mRNA for the alpha subunit was abundant in the internodes and florets, the tissues closely related to abnormality in the dwarf transformants. The position of the alpha-subunit gene was mapped on rice chromosome 5 by mapping with the restriction fragment length polymorphism. The position was closely linked to the locus of a rice dwarf mutant, Daikoku dwarf (d-1), which is known to exhibit abnormal phenotypes similar to those of the transformants that suppressed the endogenous mRNA for the alpha subunit by antisense technology. Analysis of the cDNAs for the alpha subunits of five alleles of Daikoku dwarf (d 1), ID-1, DK22, DKT-1, DKT-2, and CM1361-1, showed that these dwarf mutants had mutated in the coding region of the alpha-subunit gene. These results show that the G protein functions in the formation of normal internodes and seeds in rice. PMID- 10377458 TI - AKT3, a phloem-localized K+ channel, is blocked by protons. AB - The potassium-channel gene, AKT3, has recently been isolated from an Arabidopsis thaliana cDNA library. By using the whole-mount and in situ hybridization techniques, we found AKT3 predominantly expressed in the phloem. To study the physiological role of this channel type, AKT3 was heterologously expressed in Xenopus oocytes, and the electrical properties were examined with voltage-clamp techniques. Unlike the plant inward-rectifying guard cell K+ channels KAT1 and KST1, the AKT3 channels were only weakly regulated by the membrane potential. Furthermore, AKT3 was blocked by physiological concentrations of external Ca2+ and showed an inverted pH regulation. Extracellular acidification decreased the macroscopic AKT3 currents by reducing the single-channel conductance. Because assimilate transport in the vascular tissue coincides with both H+ and K+ fluxes, AKT3 K+ channels may be involved in K+ transport accompanying phloem loading and unloading processes. PMID- 10377459 TI - Distribution of tactile learning and its neural basis. AB - The brain's sensory processing systems are modified during perceptual learning. To learn more about the spatial organization of learning-related modifications, we trained rats to utilize the sensory signal from a single intact whisker to carry out a behavioral task. Once a rat had mastered the task, we clipped its "trained" whisker and attached a "prosthetic" one to a different whisker stub. We then tested the rat to determine how quickly it could relearn the task by using the new whisker. We observed that rats were immediately able to use the prosthetic whisker if it were attached to the stub of the trained whisker but not if it were attached to a different stub. Indeed, the greater the distance between the trained and prosthetic whisker, the more trials were needed to relearn the task. We hypothesized that this "transfer" of learning between whiskers might depend on how much the representations of individual whiskers overlap in primary somatosensory cortex. Testing this hypothesis by using 100-electrode cortical recordings, we found that the overlap between the cortical response patterns of two whiskers accounted well for the transfer of learning between them: The correlation between the electrophysiological and behavioral data was very high (r = 0.98). These findings suggest that a topographically distributed memory trace for sensory-perceptual learning may reside in primary sensory cortex. PMID- 10377460 TI - Impairments in verb morphology after brain injury: a connectionist model. AB - The formation of the past tense of verbs in English has been the focus of the debate concerning connectionist vs. symbolic accounts of language. Brain-injured patients differ with respect to whether they are more impaired in generating irregular past tenses (TAKE-TOOK) or past tenses for nonce verbs (WUG-WUGGED). Such dissociations have been taken as evidence for distinct "rule" and "associative" memory systems in morphology and against the connectionist approach in which a single system is used for all forms. We describe a simulation model in which these impairments arise from damage to phonological or semantic information, which have different effects on generalization and irregular forms, respectively. The results provide an account of the bases of impairments in verb morphology and show that these impairments can be explained within connectionist models that do not use rules or a separate mechanism for exceptions. PMID- 10377461 TI - Folkecology and commons management in the Maya Lowlands. AB - Three groups living off the same rainforest habitat manifest strikingly distinct behaviors, cognitions, and social relationships relative to the forest. Only the area's last native Maya reveal systematic awareness of ecological complexity involving animals, plants, and people and practices clearly favoring forest regeneration. Spanish-speaking immigrants prove closer to native Maya in thought, action, and social networking than do immigrant Maya. There is no overriding "local," "Indian," or "immigrant" relationship to the environment. Results indicate that exclusive concern with rational self-interest and institutional constraints do not sufficiently account for commons behavior and that cultural patterning of cognition and access to relevant information are significant predictors. Unlike traditional accounts of relations between culture, cognition, and behavior, the models offered are not synthetic interpretations of people's thoughts and behaviors but are emergent cultural patterns derived statistically from measurements of individual cognitions and behaviors. PMID- 10377462 TI - The early Upper Paleolithic human skeleton from the Abrigo do Lagar Velho (Portugal) and modern human emergence in Iberia. AB - The discovery of an early Upper Paleolithic human burial at the Abrigo do Lagar Velho, Portugal, has provided evidence of early modern humans from southern Iberia. The remains, the largely complete skeleton of a approximately 4-year-old child buried with pierced shell and red ochre, is dated to ca. 24,500 years B.P. The cranium, mandible, dentition, and postcrania present a mosaic of European early modern human and Neandertal features. The temporal bone has an intermediate sized juxtamastoid eminence. The mandibular mentum osseum and the dental size and proportions, supported by mandibular ramal features, radial tuberosity orientation, and diaphyseal curvature, as well as the pubic proportions align the skeleton with early modern humans. Body proportions, reflected in femorotibial lengths and diaphyseal robusticity plus tibial condylar displacement, as well as mandibular symphyseal retreat and thoracohumeral muscle insertions, align the skeleton with the Neandertals. This morphological mosaic indicates admixture between regional Neandertals and early modern humans dispersing into southern Iberia. It establishes the complexities of the Late Pleistocene emergence of modern humans and refutes strict replacement models of modern human origins. PMID- 10377463 TI - Canadian Institutes of Health Research: should there be an institute for digestive sciences? PMID- 10377464 TI - Reduction in morbidity and mortality associated with gastrointestinal bleeding in the elderly. PMID- 10377465 TI - Role of high fibre foods in the prevention of colorectal neoplasia. PMID- 10377466 TI - Retiring at 55: is it a media-created myth? PMID- 10377467 TI - Barrett's esophagus: is it all that bad? AB - An alarmingly rapid rise in the number of adenocarcinomas at the level of the gastroesophageal junction and distal esophagus has been noted over the past two decades. Intestinal metaplasia is considered to be the main precursor lesion for such adenocarcinomas. Given the low five-year survival rate in patients with advanced esophageal cancer, strategies for early detection have been developed. Because superficial cancers only rarely cause symptoms, detection of cancer at such an early curable state can only be achieved through surveillance of patients at risk. Therefore, implementation of an endoscopic surveillance program for patients in whom intestinal metaplasia has been detected in the distal esophagus or at the esophagogastric junction seems to be a reasonable option. PMID- 10377468 TI - New photosensitizers for photodynamic therapy in gastroenterology. AB - Most applications of photodynamic therapy (PDT) in gastroenterology to date have used porfimer sodium as the photosensitizing agent. For destroying small lesions in the wall of the gastrointestinal tract in inoperable patients, it has proved to be most effective, but attempts to achieve circumferential mucosal ablation, as in the treatment of Barrett's esophagus, have led to a high incidence of strictures, and all patients have cutaneous photosensitivity, which can last up to three months. Two new photosensitizers are of particular interest to gastroenterologists. PDT with metatetrahydroxyphenyl chlorin produces a similar biological effect as PDT with porfimer sodium, but the light doses required are much smaller, and cutaneous photosensitivity lasts only two to three weeks. Further, it can be used with percutaneous light delivery to destroy localized pancreatic cancers. The photosensitizing agent 5-amino levulinic acid, converted in vivo into the photoactive derivative protoporphyrin IX, sensitizes the mucosa much more than the underlying layers. This makes it feasible to destroy areas of abnormal mucosa without damaging the underlying muscle and is, therefore, better for treating Barrett's esophagus. Detailed clinical studies are required to establish the real role of PDT with the use of these and other new photosensitizers. PMID- 10377469 TI - Results of photodynamic therapy in Barrett's esophagus: A review. AB - Barrett's esophagus is associated with an increased occurrence of mucosal dysplasia and adenocarcinoma in the specialized glandular mucosa, with a 30- to 52-fold increase in the occurrence of esophageal cancer compared with the normal population. An alternative to esophagectomy as a treatment modality is needed because of the high morbidity and mortality associated with it. Photodynamic therapy offers an alternative nonsurgical therapy that eliminates dysplasia and superficial cancer, and reduces Barrett's mucosa while reducing the risks and costs compared with those of esophagectomy. The use of photodynamic therapy in the ablation of Barrett's mucosa is reviewed. PMID- 10377470 TI - Colorectal cancer screening: A guide to the guidelines. AB - The two most recent guidelines for colorectal cancer screening are those of the Agency for Healthcare Policy and Research, and the American Cancer Society. The guidelines are similar in many regards and reflect current literature, consensus opinion and compromise between members of multidisciplinary panels. The emphasis of both guidelines is to increase the options available for colorectal cancer screening. Increasing choice should expand the attractiveness of colorectal cancer screening to more patients and physicians, and the development of guidelines should help compel payers to provide reimbursement for colorectal cancer screening. These guidelines are summarized and evaluated as they pertain to colorectal cancer screening. PMID- 10377471 TI - Zapping Zenker's diverticulum: gastroscopic treatment. AB - Zenker's diverticulum (ZD) is a common cause of dysphagia in the elderly. Many symptomatic elderly are poor candidates for surgery and/or ear, nose and throat treatment. The author's first experiences with gastroscopic treatment by cutting the Zenker bridge to allow an overflow have recently been published. Only patients with contraindications for general anesthesia were accepted to the pilot group. However, the author now treats all ZD patients in this manner. One hundred and twenty-five patients (male to female ratio 1. 6) were referred for treatment from 1993 to 1997. After introduction of the gastroscope into the esophagus, a nasogastric tube was positioned to treat a ZD bridge with a height of less than 1 cm. The ZD bridge was divided by argon plasma coagulation, if necessary, in combination with monopolar forceps, Savary dilator and/or precut needle. All patients received antibiotics, topical anesthesia to the throat, if necessary, and intravenous midazolam, if possible. Radiography was performed after treatment. Normalization of the diet was allowed when the x-ray showed no signs of leakage. All patients referred for treatment were treated successfully. The median age was 77 years (range 41 to 100 years). Symptomatic improvement was seen in all patients after treatment. Complications included subcutaneous emphysema (n=17), mediastinal emphysema (n=5) and bleeding (n=2). One patient (95 years of age) died in her nursing home 27 days after treatment due to massive pulmonary embolism. The thirty-day mortality rate was otherwise zero. Three patients had been previously treated by surgeons and 12 by ear, nose and throat physicians, without sufficient improvement; all were adequately treated by the author. The mean number of treatment sessions was 1.8. This approach seems safe and effective. Treatment of every patient was possible and was carried out, even in patients in very poor condition, without general anesthesia. PMID- 10377472 TI - Preventing ulcer rebleeding: the role of second-look endoscopy. AB - Whether a second-look endoscopy after initial endoscopic hemostasis is of value is controversial. Routine surveillance endoscopy on the next day and treatment of any remaining stigmata may provide marginal benefit, but existing studies are not large enough to demonstrate significant differences. Endoscopic retreatment when patients develop rebleeding may allow emergency surgery to be avoided if successful but may endanger life if further bleeding occurs. Early data indicate that surgery can be avoided in about 75% of patients by retreatment without an increase in mortality. PMID- 10377473 TI - Spots and clots - leave them or treat them? Why and how to treat. AB - The clinical indication for urgent endoscopy with combined diagnosis and treatment is bleeding that is severe enough to seek medical attention. The author uses stigmata of ulcer hemorrhage as a guide to endoscopic therapy. Active arterial bleeding, nonbleeding visible vessels and adherent nonbleeding clots are always treated endoscopically. In randomized trials, patients have demonstrated better outcomes from endoscopic therapies than from medical therapies. Flat spots, grey or black sloughs, and clear ulcer bases are not treated endoscopically. The clinical condition and the endoscopic appearance of the ulcer (ie, stigmata of hemorrhage) of the patient with upper gastrointestinal bleeding are used to determine the subsequent level of care (discharge, ward or intensive care). PMID- 10377474 TI - Is light-induced fluorescence better than the endoscopist's eye? AB - While recognizing advanced tumours with endoscopy does not generally pose a challenge, cure rates are relatively low, depending on the size and stage of the tumour. Screening tests for cancer are advantageous for diagnosing cancers before the date after which a cure is no longer an option. Many gastrointestinal cancers are diagnosed after the date on which a cure is possible. The present article discusses some of the limitations of conventional white light endoscopy in screening and presents some of the fluorescent-based diagnostics that are being investigated as complements to white light endoscopy. Autofluorescence and fluorescence due to exogenous photosensitizers or precursors are two sources of fluorescence that are being studied. Preliminary results of current investigations are presented, and future research directions are described. PMID- 10377475 TI - Tissue staining (chromoscopy) of the gastrointestinal tract. AB - Tissue staining, or chomoscopy, is used as an adjunctive technique during gastrointestinal endoscopy. Chemical agents are applied to the gastrointestinal mucosal surface to identify specific epithelia or to enhance the mucosal surface characteristics of the gastrointestinal epithelium. This aids in the recognition of subtle lesions (ie, polyps) or allows directed targeting of biopsies (ie, sprue or Barrett's esophagus) to increase the yield of endoscopic diagnostic accuracy. The four endoscopic tissue-staining techniques in use are vital staining, contrast staining (chromoscopy), reactive staining and tattooing. Some of the agents used for endoscopic tissue staining and the uses of chromoscopy in identifying pathology of the esophagus, stomach, small bowel and colon during endoscopy are discussed. PMID- 10377476 TI - [Pathophysiologic aspects of S-100beta protein: a new biological marker of brain pathology]. AB - S-100 protein is an acidic calcium-binding protein with a molecular weight of 21 kDa consisting of two submits alpha and beta, which are combined to give alphaalpha (S-100a), alphabeta (S-100a) and betabeta (S-100b) dimeric forms. S 100 protein is much more abundant in the brain than in other tissus and is present as a mixture of S-100a and S-100b (named S-100beta). These two isoforms are predominantly synthesized and secreted by glial cells. Structural damage of these cells causes leakage of S-100beta protein into the extracellular compartment and into cerebrospinal fluid, further entering the bloodstream. We provide here an overview of the physicochemical properties of S-100beta protein, of its pathological aspects and of possible interest in measuring this protein in biological fluids during cerebral diseases and brain damage occurring after surgical events. PMID- 10377477 TI - [Carotenoids: 2. Diseases and supplementation studies]. AB - Inverse correlations have been found in most studies on the relationship between dietary intake and plasma concentrations of carotenoids on one side and degenerative diseases such as cancer and cardiovascular diseases on the other side. Protective effects of carotenoids have been found for pathologies of the retina and the skin. Concentrations of these molecules in blood are lower in digestive pathologies and HIV. Short- and long-term toxicity of carotenoids was found to be low. In combination with the beneficial effects found for diets rich in carotenoids, this has initiated trials with relatively high doses of carotenoid supplements. In the study in Linxian (China) in a rural population with poor nutritional status, supplementation with beta-carotene, zinc, selenium and vitamin E lowered total mortality and mortality from stomach cancer. Other studies (ATBC, Caret.) on well-fed subjects did not show beneficial effects on mortality from cancer and cardiovascular diseases. On the contrary, higher mortality and lung cancer incidence was found in supplemented subjects that were also exposed to asbestos and cigarette smoke. In these studies, doses of supplemental beta-carotene were high and varied from 20 to 50 mg/day. One still ongoing study, called Suvimax, doses subjects for eight years with a cocktail of vitamins and minerals including 6 mg per day of beta-carotene. This supplementation with physiologically seen more "normal" doses might give clarity on the question if beta-carotene is the protective factor in fruits and vegetables. PMID- 10377479 TI - [Viruses and the neuroendocrine system: model of murine obesity induced by cerebral infection by canine distemper virus]. AB - It is currently well established that the nervous, endocrine and immune systems inter-communicate using biologically active soluble factors, synthesised and produced by these three systems themselves (e.g. immunomodulator effect of hormones, effect of substances secreted by immune cells on endocrine function.). In addition, these systems jointly express receptors for hormones, peptides, growth factors and cytokines. Immuno-neuroendocrine interactions therefore underlie physiological processes and their deregulation can result in various pathological states. By entering into complex relationships with the specialized and differentiated cells of these three systems viruses can alter inter-cellular communication and result in the appearance of pathological processes directly linked to these disturbances. In order to understand the role of viruses in the genesis of neuroimmunoendocrine pathologies, we have developed a cerebral infection model using canine distemper virus (CDV). In infected mice, this paramyxovirus, closely related to the human measles virus, induces early neurological pathologies (encephalitis) which are associated with active viral replication. Mice surviving the acute phase of infection exhibit motor deficits (paralysis and turning behaviour) or obesity during the viral persistence phase, despite the fact that the virus is no longer detectable. The obesity is characterised by hyperinsulinaemia, hyperleptinaemia and hyperplasia of the adipocytes, associated with decreased expression of the OB-Rb hypothalamic leptin receptor and modulated expression of hypothalamic monoamines and neuropeptides. These results support the viral "hit and run" theory, since the initial viral impact in the hypothalamus may be the origin of the changes in later immunoneuroendocrine communication. Thus, certain human neurodegenerative or neuroendocrine diseases may have a previous viral infection aetiology without it being possible to clearly identify the agent responsible. PMID- 10377478 TI - [Anti-viral proteins: from interferon alpha to its receptor]. AB - The immune response against viral pathogens include specific and non specific mechanisms. Cytokines are peptides which can play a role in the non specific immunity. Type I interferons (IFNalpha/beta) are the most effective antiviral cytokines. Interferons alpha are represented by a large familly of structurally related genes while the IFNbeta is encoded by a single gene. Type I interferon genes are located on the chromosome 9, and are segregated in a "modern" and an "ancestral" group with distinctive effects onto the cells. Type I interferons consist of 5 alpha helices, 4 of which are closely held together. Type I interferons receptor is a member of the class II cytokine receptor familly. It is a heterodimer composed of polypeptidic chains naimed IFNAR-1 and IFNAR-2. The type I interferons inducers are viruses, inactivated viruses and certain synthetic molecules. The molecular mechanisms of the induction of IFNbeta have been extensively described whereas the factors that play a role in the regulation of IFNalpha are not so well characterized. Two regulating domains located on the IFNbeta gene promotor are involved. The activation of these regulating domains leads to adverse effects by interacting with two intracellular factors (IRF1 and IRF2). IRF1 enhances the synthesis of IFNbeta but other pathways may be involved as well. The different IFNalpha sub-types have not only synergistic but also antagonistic effects. The synthesis of the different subtypes depends on cell lines and IFNalpha inducers. The N terminal part of IFNalpha is a major determinant in the diversity of the biological effects of IFNalpha sub-types which probably involve changes of the conformation of the type I receptor resulting from interactions between the receptor and its ligand. The type I interferons confer resistance to viruses, especially by enhancing the synthesis of intracellular antiviral proteins. PMID- 10377480 TI - [Genetic causes of male infertility]. AB - In this review, after a brief summary of the spermatogenesis process, we present some genetic causes of male infertility at chromosome and gene level, known at present. Even though knowledge has greatly increased, in 15% of cases, we still do not know the infertility mechanism. The genotype-phenotype relationship remain ambiguous because of the diversity of the implicated mechanisms and their interactions. Other genes remain to be identified and, in the future, one of the main difficulties will be to define exactly the relationship between environmental factors and male infertility. PMID- 10377481 TI - [Genetic control of spermatogenesis: Y chromosome and male infertility]. PMID- 10377482 TI - [Physiopathology of gonadotropic control of gametogenesis]. PMID- 10377483 TI - [Fertility of the couple and fecundity: genetic aspects]. PMID- 10377484 TI - [Natural reproduction and assisted reproduction]. PMID- 10377485 TI - [Diagnostic inquiry into hyperprolactinemic pathologies]. PMID- 10377486 TI - [The spermogram and the exigencies of quality]. PMID- 10377487 TI - [The Paragon CZE 2000 and the Hydrasys Hyrys for electrophoresis of serum proteins and serum monoclonal antibody typing]. PMID- 10377488 TI - [Comparison of LDL-cholesterol results calculated by using apolipoproteins A1, B or HDL-cholesterol for classification of non-chylomicron dyslipidemias]. PMID- 10377489 TI - [Evaluation of analytical performances of ferritin assays with Vitros ECi]. PMID- 10377490 TI - [A surprising aglycorrachia! A case of neurocysticercosis]. PMID- 10377491 TI - [Program of external quality control in cyclosporine dosage: summary and analysis from French laboratories]. PMID- 10377492 TI - [Evaluation of a new automated method for von Willebrand factor antigen measurement: the STA-Liatest vWF]. PMID- 10377493 TI - [Selective salpingography in the treatment of infertility caused by proximal tubal obstruction: apropos of 122 cases treated in Dakar]. AB - One hundred and twenty-two women who had not conceived after more than one year of unprotected sexual relations underwent selective salpingography. All had been shown to have proximal obstruction of the fallopian tube by standard hysterosalpingography. The women underwent the procedure as outpatients during the follicular period. They were given prophylactic antibiotic treatment and salpingography was performed by a radiologist in an X-ray room. We used the commercial sets of Cook and Zorn. The mean age of patients was 34 years (range 28 to 46) and the mean duration of infertility was 7.7 years (range 2.2 to 14 years). Two hundred and forty tubes were examined, of which 213 were cleared of obstruction (88.7%), with both tubes affected in 66% of cases and only one tube affected in 33% of cases. All patients had at least one patent tube after the procedure. The patent tube was normal in 73% of cases (177 tubes) and abnormal in 15% (36 tubes). Catheterization failed in 11.3% of cases (27 tubes). Forty-nine women conceived spontaneously during the 2- to 12-month follow-up period, and 39 of these women delivered healthy babies. No extrauterine pregnancies or serious complications were reported. The most frequent side effects were slight pain, bleeding and nausea. One perforated tube was reported and none of the patients died. The greatest difficulties encountered were a lack of cooperation in patients with previous painful experiences of hysterogram and catheterization problems in women with a strongly flexed or distorted uterus. Complications were minimal and the one case of tubal perforation at the beginning of the series had no serious consequences. Thus, selective salpingography should be used more widely because it is simple and more cost-effective than the surgical management of tubal obstruction and artificial insemination. Our results suggest that this procedure should be the first-line treatment of infertility caused by proximally obstructed fallopian tubes. PMID- 10377494 TI - [Ultrasound aspects of abdominal involvement in adults with HIV infections in the Ivory Coast: apropos of 146 cases]. AB - Over a six-month period, 146 grade IV AIDS patients (HIV-1 and HIV-2) systematically underwent abdominal ultrasound examination. All abdominal organs were investigated in the search for lesions. Many lesions were detected. Pathological lesions were most frequent in the nodes (17.2%), hyperechogenic liver tissue (16.5%) and kidneys (13.7%). Lesions in the liver, gall bladder and spleen were less frequent. In most cases, the lesions were not specific to AIDS. However, systemic abdominal ultrasound scans for AIDS patients would be of value because such lesions are present. PMID- 10377495 TI - [Visual problems in albinos: a hospital study carried out at the Douala General Hospital]. AB - We report here a clinical study investigating the main visual problems associated with albinism, with a view to determining the best treatment. We came across 42 cases of albinism during the course of this study, corresponding to a prevalence for albinism of 0.15%. One in ten albinos were of the yellow mutant type and more of the albinos were men than were women (sex ratio 1.21). The maximum visual acuity recorded was 3/10 and 40.47% of the patient had a best visual acuity score no higher than 1/10. Vision was best in the yellow mutants and it improved with age. We found that 33.33% of the albino patients had a squint and 35.71% had torticullis at primary fixation. In contrast to the results of previous studies, the most common ametropia was myopic astigmatism (61.9%). These findings have potential implications for the treatment of visual problems in albinos. PMID- 10377496 TI - [The consumption of cassava is not responsible for the etiology of endemic goiter in rural areas in Senegal]. AB - An epidemiological study of iodine deficiency disorders (IDD) was carried out in West Senegal and Casamance. Five hundred and eighty five households were selected in 3 areas with a high prevalence of goiter. We assessed the relative impact of iodine deficiency (estimated by mean iodine excretion in the urine of family members) and cassava consumption (mean frequency of consumption by the household). Cassava consumption, even if on a regular basis, neither caused nor increased goiter formation in this area of West Africa. This was probably due to the local method of cassava root preparation, which reduces the amount of cyanogenic compounds consumed. Iodine deficiency was principally responsible for goiter formation. Therefore, the commercial availability of iodine-supplemented salt should lead to the eradication of IDD from Senegal in the near future. PMID- 10377497 TI - [Sexual behavior, knowledge and attitudes to AIDS and sexually transmitted diseases of students at the University of Benin (Togo)]. AB - Many studies have shown that in Africa, particularly in Togo, the 20- to 29-year old age group is the age group most frequently affected by AIDS. This age group accounts for 84% of the students of the University of Benin. We studied students, most of the age group thought to be most at risk, investigating sexual behavior, knowledge and attitudes to AIDS and sexually transmitted diseases (STDs). The level of knowledge about the problems of AIDS and STDs was similar for both sexes and for all ages and levels of education of the students. Students had a reasonable knowledge of AIDS, particularly concerning the transmission of HIV (88.6% of students aware), risk behavior (80.8%), AIDS treatment (57.0%) and more general information about HIV (49.4%). They were poorly informed about the transmission (42.9%) and complications (0.69%) of other STDs. Most students had positive attitudes towards HIV issues, particularly the use of preventive measures (3.41 in 5) and the acceptance of infected individuals (3.98 in 5). However, few had seriously considered that AIDS and STDs might impact on their own sex lives (1. 84 in 5) and some were even fatalistic concerning HIV infection. The students were highly sexually active, having intercourse a mean of 31 times per year. Their sexual behavior depended on age and sex. The 15- to 19 year-olds preferred occasional partners. They had sexual intercourse 1 to 3 times per month and used condoms 10 to 20% of the time. The 20- to 29-year-olds had multiple partners. They had sexual intercourse 3 to 5 times per month and used condoms more than 30% of the time. Students over the age of 30 had many partners in addition to their regular partner. They had sexual intercourse 5 to 10 times per month and used condoms 0 to 20% of the time. Significantly more women than men had high-risk sexual behavior (40. 5% of men claimed to regularly use condoms, versus only 22.7% of the women and 11.9% of the women accepted anal penetration versus only 8. 4% of men). The general assumption is that students, who have a high level of education, should be well informed concerning AIDS and STDs and should therefore have positive attitudes and responsible sexual behavior. This study demonstrates that the assumption bears no resemblance to reality. The students were aware of the way in which HIV is transmitted and of what construes risky behavior, but there was nonetheless a high frequency of high risk behavior (e.g. multiple sexual partners, anal and oral sex, homosexuality, intravenous drug use). The behavior of the students was not significantly different from that of young people living on the streets of Lome. There were significant relationships between knowledge and attitudes and between some types of sexual behavior and knowledge. However, there was no correlation between attitudes and behavior. The education of young people should focus on both the prevention of AIDS and STDs and on facing up to these diseases. PMID- 10377499 TI - [The time is ripe for listening to the voice of the half of the world]. AB - Has the world of women changed? So wonders a one-time champion of what was then called women's rights, and is now known as women's empowerment, in the Third World. She recalls her surprise when she listened to advocates of birth spacing as a young mother. She remembers the debates she attended: "Today development, tomorrow the Pill", "Women's education is the key to progress", the speculations of western feminists ignorant of the burden of traditions kept alive elsewhere. She is now concerned about the resolution of the Beijing conference, assigning the protection of the environment and responsibility for domestic husbandry to women. Is this not yet another abdication of male responsibility? Should she once again stand up and deliver a harangue at an international meeting or should she instead pursue the quiet, self-effacing weaving of networks between the women of the North and South? PMID- 10377498 TI - [Use in Mozambique of WHO diagnostic strategies in HIV infection]. AB - The lack of expensive equipment and well-trained laboratory technicians in developing countries makes it difficult to use standard methods (2 EIA confirmed by western blotting) to diagnose HIV infection. This led the WHO to develop simplified algorithms based on sequential screening tests, with no confirmation by immunoblotting. These algorithms were tested in the normal diagnosis conditions of a medical unit in Maputo, Mozambique. We tested 402 sera, collected with the consent of the patients concerned. The sera were first tested for HIV according to French regulations (2 EIA with western blot if at least one EIA was positive). This strategy identified 53 sera as positive for HIV1 and 1 serum as positive for HIV2. We then tested who algorithms, one for a predicted rate of prevalence < 10% and the other for a predicted rate of prevalence > 10%. Neither algorithm performed adequately for diagnostic purposes. Further evaluation with a panel of local sera is required to select the most suitable algorithm in terms of specificity and sensitivity, and algorithms should be throughly tested before inclusion in national AIDS control strategies. PMID- 10377500 TI - [Amebic liver abscesses in Yaounde]. AB - Until 1985, morbidity and mortality from amebic infections were high in sub Saharan Africa (2). The aim of this work was to describe the clinical, diagnostic and prognostic aspects of amebic liver lesions in a large town in tropical Africa, seven years after the last study of this disease in the area. We studied 96 amebic liver abscesses in 77 patients in a study with prospective and retrospective components. The patients were aged 20 to 65 years and the diagnoses were made on the basis of clinical radiological and biological criteria. Most of the patients were men (3:1). The most frequent clinical signs and symptoms were: pain on (100%), fever (92.2%), enlarged right hypochondrium (89.6%), heptomegaly (45.4%) and weight loss (39.6%). Ultrasound scans showed that the abscesses were most frequently found in the liver, on the right-hand side (65.6%) and that they were 20 to 125 mm in diameter. Between 80 and 2,500 ml of pus was drained from each abscess. We found pleuropulmonary lesions in 10.4% of cases. Serological tests for amebae were strongly positive in almost all cases and HIV tests (carried out prospectively) were positive in 11.6% of cases. The amebic liver lesions appeared to be primary in 66% of cases. The patients were treated with metronidazole, combined if necessary, with drainage under ultrasound surveillance. Two of the patients died. The others were completely cured after a mean of 13 days in hospital. Amebic infestations are cosmopolitan in nature. They occur most frequently in tropical areas. Amebic infections are rife in tropical Africa, with a prevalence of 1 to 2% (1). Liver amebiasis is the principal form of amebic infection outside of the intestine. Liver amebiasis is often detected at the stage of abscess formation. PMID- 10377502 TI - [Ethical issues and preventive vaccination]. AB - Preventive vaccination is one of the most spectacular examples of success in the fight against disease. However, the implementation of vaccination strategies is not dependent solely on identifying appropriate vaccines. It also requires organization and is affected by complex economic issues, which are subject to ethical considerations. These include the mobilization of resources and skills for the common good and the respect of the individual such that the use and acceptability of vaccines are optimized. PMID- 10377501 TI - [CISMeF: catalog and index of French-speaking medical sites]. AB - The Internet has now become a major source of health information. The aim of CISMeF is to catalogue and index the main French-speaking sites and documents concerning health. This project was initiated by Rouen University Hospital. Its URL is http://www.chu-rouen.fr/cismef. CISMeF covers all areas of health care and medical sciences, and is indexed both alphabetically and according to subject. It was set up on a Sun workstation under the Sun UNIX operating system and is entirely based on static HTML. By May 1999, the number of sites and documents indexed was already over 6,500, with a mean of 75 new sites added each week. CISMeF is updated via a five-step process: resource collection, filtering, description, classification, and indexing. The Net Scoring criteria are used to assess the quality of health information on the Internet. These criteria concern eight categories: credibility, content, links, design, interactivity, quantitative aspects, ethics and accessibility. CISMeF uses two standard tools to organize information: the MeSH (medical subject heading) thesaurus from the Medline reference database (National Library of Medicine, USA) and the Dublin core metadata format. The sites and documents included in CISMeF are described using the following elements from the Dublin core project: title, author or creator, subject and keywords, description, publisher, date, resource type, format, identifier, and language. PMID- 10377503 TI - Octreotide suppresses the incretin glucagon-like peptide (7-36) amide in patients with acromegaly or clinically nonfunctioning pituitary tumors and in healthy subjects. AB - OBJECTIVE: To study the effect of octreotide on glucagon-like peptide (7-36) amide (GLP-1) and insulin secretion in patients with pituitary tumors during preoperative treatment and in healthy subjects. DESIGN: Open design prospective clinical study. METHODS: Eighteen patients with pituitary macroadenomas (13 clinically nonfunctioning (NFA; 11/13 had GH insufficiency), 5 GH secreting (GHA)) received preoperative octreotide treatment: 3x100 microg/day s. c. for 3 months, and 3x500 microg/day s.c. for an additional 3 months. Seven healthy subjects received (for ethical reasons) only 3x100 microg/day for 10 days. A standardized meal (St-M) test, oral glucose test (oGTT) and i.v. glucose test (ivGTT) were done before octreotide therapy, on days 1, 2 and 3 (D1,2,3), after 3 months (M3) and 6 months (M6) of octreotide treatment in the patients, and before treatment, on D1,2,3 and on D8,9,10 of octreotide treatment in the healthy subjects. Serum GLP-1, insulin and GH as well as plasma glucose were determined for 180 min (oGTT, St-M) or 120 min (ivGTT). RESULTS: Pretreatment fasting GLP-1 concentrations as well as integrated responses (area under the curve 0-180 min) to oGTT and St-M were not significantly different between NFA, GHA and healthy subjects. During the oGTT, octreotide initially almost abolished the early (0-60 min) and diminished the late (60-180 min) GLP-1 and insulin responses in patients and healthy subjects. At M6 integrated insulin responses had significantly recovered, while the increase in GLP-1 response failed to reach significance (GLP 1: 56.5% of pretreatment at D2 versus 93.5% at M6 and 41.2 versus 63.1% in NFA and GHA respectively; insulin: 50.2 versus 71.2% and 35.5 versus 70. 4%). An escape of GLP-1 and insulin in healthy subjects (D2 versus D9) was not significant. Intestinal glucose absorption was apparently not reduced, since the early glucose rise was similar before and during octreotide treatment. During the St-M the GLP-1 and insulin responses were similarly suppressed by octreotide and recovered during ongoing treatment (GLP-1: 49.6% of pretreatment at D1 versus 79.0% at M6 in NFA and 46.9 versus 52.9% in GHA. Insulin: 27.6 versus 83.9% and 23.5 versus 54.4%). The escape was significant in NFA but not in GHA. In the healthy subjects the escape was already significant on D8 (GLP-1: 39.5% of pretreatment at D1 versus 68.3% at D8; insulin: 36.6 versus 53.8%). During the ivGTT GLP-1 did not increase. The early insulin response (0-30 min) was abolished by octreotide, followed by a reduced peak at 60 min. The reduction of the integrated insulin response during ivGTT was similar to that during oGTT. An insulin escape reached significance only for NFA (52. 6% of pretreatment at D3 versus 66.7% at M6). Glucose tolerance (KG value) deteriorated and did not improve during ongoing treatment. Octreotide suppressed the median GH concentration (8h profile) of the GHA patients from 10.3 microg/l (pretreatment) to 5.8, 6.3 and 3. 7 microg/l at D4, M3 and M6 with no escape. GH was 1.5 microg/l postoperatively. CONCLUSIONS: Octreotide abolishes the early and diminishes the late GLP-1 and insulin responses to oGTT and St-M in NFA and GHA patients and in healthy subjects. In contrast to GH, both hormones partially escape from suppression during ongoing therapy. During treatment with our conventional octreotide doses suppression of insulin secretion is maximal. Under these conditions an effect of the additional loss of GLP-1 is not apparent. Basal GLP-1 concentrations and integrated responses to oGTT and St-M were similar in healthy subjects and in patients with GH excess or GH insufficiency. PMID- 10377505 TI - Biotechnology clusters. PMID- 10377504 TI - Effects of 42 months of GH treatment on bone mineral density and bone turnover in GH-deficient adults. AB - OBJECTIVE: To study the effects of GH treatment for up to 42 months on bone mineral density (BMD) and bone turnover. DESIGN AND METHODS: BMD with dual energy X-ray absorptiometry, serum type I procollagen carboxy-terminal propeptide (PICP), serum type I collagen carboxy-terminal telopeptide (ICTP) and serum IGF-I were assessed in 71 adults with GH deficiency. There were 44 men and 27 women, aged 20 to 59 (median 43) years. Thirty-two patients completed 36 months and 20 patients 42 months of treatment. RESULTS: The BMD increased for up to 30-36 months and plateaued thereafter. In the whole study group, the maximum increase of BMD was 5.0% in the lumbar spine (P<0. 001), 5.9% (P<0.01) in the femoral neck, 4.9% (NS, P>0.05) in the Ward's triangle and 8.2% (P<0.001) in the trochanter area. The serum concentrations of PICP (202.6+/-11.5 vs 116.3+/-5.4 microg/l; mean+/-s.e.m.) and ICTP (10.5+/-0.6 vs 4.4+/-0.3 microg/l) doubled (P<0.001) during the first 6 months of GH treatment but returned to baseline by the end of the study (130.0+/-10.4 and 5.6+/-0.7 microg/l respectively), despite constantly elevated serum IGF-I levels (39. 6+/-4.1 nmol/l at 42 months vs 11.9+/ 0.9 nmol/l at baseline; P<0.001). The responses to GH treatment of serum IGF-I, PICP, ICTP (P<0.001 for all; ANOVA) and of the BMD in the lumbar spine (P<0.05), in the femoral neck and the trochanter (P<0.001 for both) were more marked in men than in women. At the end of the study the BMD had increased at the four measurement sites by 5.7-10.6% (P<0.01-0.001) in patients with at least osteopenia at baseline and by 0.1-5.3% (NS P<0.05) in those with normal bone status (P<0.001 for differences between groups; ANOVA). Among patients who completed 36-42 months of treatment, the number of those with at least osteopenia was reduced to more than a half. The response of BMD to GH treatment was more marked in young than in old patients at three measurement sites (P<0. 05-<0.001; ANOVA). In the multiple regression analysis the gender and the pretreatment bone mass appeared to be independent predictors of three measurement sites, whereas the age independently determined only the vertebral BMD. CONCLUSIONS: GH treatment in GH-deficient adults increased BMD for up to 30-36 months, with a plateau thereafter. Concurrently with the plateau in BMD the bone turnover rate normalized. From the skeletal point of view GH-deficient patients exhibiting osteopenia or osteoporosis should be considered as candidates for GH supplementation of at least 3-4 years. PMID- 10377506 TI - Chemokines and their receptors. PMID- 10377507 TI - Biotechnology perspectives in Europe. PMID- 10377508 TI - Green fluorescent protein (GFP): applications in cell-based assays for drug discovery. AB - Green fluorescent protein (GFP) is a powerful tool for cell-based assays owing to the intrinsic fluorescence of this protein that allows real-time analysis of molecular events in living cells. A number of GFP variants have been developed with optimal properties for both high-throughput screening and high-content screening. The author discusses advances in basic GFP technology, including the discovery of fluorescent proteins from divergent bioluminescent species, as well as the development of various GFP biosensors suited to the drug discovery process. PMID- 10377509 TI - Intellectual property and chirality of drugs. AB - Chirality has emerged as a key issue in drug design, discovery and development. Chiral switches are drugs that are already approved and claimed as racemates but that have been redeveloped as single enantiomers. The legal state of the art of patentability of chiral switches, as derived from US and European precedents, is reviewed. The issues of intellectual property in the pending chiral switches of the blockbuster drugs ibuprofen, fluoxetine and omeprazole are analysed. PMID- 10377510 TI - Dopaminergic agents for the treatment of cocaine abuse. AB - Cocaine is a major drug of abuse whose devastating effects have captured the attention of health officials and policy makers. Based upon the alarming health and crime-related costs associated with the use of this powerful reinforcing drug, immediate therapies are needed for the treatment of cocaine addiction. In this review, some of the small-molecule-based approaches that have been pursued in the search for such medications are highlighted. Because the pharmacological actions of cocaine stem laargely from its ability to block the dopamine transporter, many intervention strategies have focused on the dopaminergic pathway. PMID- 10377511 TI - Monitor: molecules and profiles. AB - Monitor provides an insight into the latest developments in drug discovery through brief synopses of recent presentations and publications together with expert commentaries on the latest technologies. There are two sections: Molecules summarizes the chemistry and the pharmacological significance and biological relevance of new molecules reported in the literature and on the conference scene; Profiles offers commentary on promising lines of research, emerging molecular targets, novel technology, advances in synthetic and separation techniques and legislative issues. PMID- 10377512 TI - Combinatorial chemistry. PMID- 10377513 TI - A step towards gene therapy for CF. PMID- 10377514 TI - New collaborations make pharmacogenomics a SNP. PMID- 10377515 TI - A vaccine approach to healthy lipoprotein levels. PMID- 10377516 TI - Rapid update PMID- 10377517 TI - IL-6 and gp130 signalling: its role in chronic disease. PMID- 10377518 TI - Procalcitonin--a novel biochemical marker for the mediator-directed therapy of sepsis. PMID- 10377519 TI - Why do in utero stem cell transplants sometimes fail? AB - In utero stem cell transplantation promises a novel therapeutic approach to those genetic disorders that can be diagnosed in early pregnancy and that could lead to either severe disability or death. Available scientific evidence suggests that such procedures could achieve clinically relevant levels of engraftment with donor cells and that the resulting sustained chimerism is potentially long lived. However, the relatively few cases performed so far have not borne out initial hopes, and both the source and the type of cell preparation to be transplanted and the choice of disorders to target with this therapy remain controversial. PMID- 10377521 TI - Gene therapy using HVJ-liposomes: the best of both worlds? AB - A new concept for the development of novel vectors is to overcome the limitations of individual vectors by combining them. The HVJ-liposome was developed by combining liposomes with fusion proteins derived from the hemagglutinating virus of Japan (HVJ), also known as Sendai virus. Gene transfer in vivo using this delivery system can be repeated because it is much less immunogenic and cytotoxic than other viral-vector systems. By coupling the Epstein-Barr virus (EBV) replicon apparatus with HVJ-liposomes, transgene expression can be sustained in vitro and in vivo. In animal models, this system has shown promise for several diseases, including cancer and cardiovascular disease. PMID- 10377520 TI - Antibiotic peptides from higher eukaryotes: biology and applications. AB - Gene-encoded antibiotic peptides are increasingly being recognized as effector molecules of host defense in plants and animals. Studies of antimicrobial peptides are providing new insights into the dynamic interactions between microbes and their hosts, and are generating new paradigms for the pathogenesis and treatment of diseases. Because antimicrobial peptides of higher eukaryotes differ structurally from conventional antibiotics produced by bacteria and fungi, they offer novel templates for pharmaceutical compounds that could be effective against increasingly resistant microbes. PMID- 10377522 TI - The putative role of cell adhesion molecules in endometriosis: can we learn from tumour metastasis? AB - Endometriosis, one of the most frequent diseases in gynaecology, is a considerable threat to the physical, psychological and social integrity of women. The etiology and pathogenesis of this important disease, defined as the ectopic location of endometrium-like glandular epithelium and stroma outside the uterine cavity, is poorly understood. Clinical observations and in vitro experiments imply that endometriotic cells are invasive and able to metastasize. Analogous to tumour metastasis, it is likely that cell adhesion molecules are central for the invasion and metastasis of endometriotic cells. Investigation of these molecules in endometriosis should increase our understanding of the molecular mechanisms involved in the pathogenesis of this disease. PMID- 10377523 TI - Alcohol and genetics: new animal models. AB - In recent years, it has become increasingly evident that there is a genetic component to alcoholism. Attempts to isolate alcoholism genes have met with modest success, in part because alcoholism is a multigenic trait. Recently, experimental animal models and novel genetic manipulations have provided several clues as to the specific genes involved in alcoholism, and extensive research has identified many genes that might influence responses to alcohol. Although not all of these might be proven to influence drug sensitivity, research has provided evidence for the involvement of a few genes. Ultimately, findings from animal models that investigate the function of specific genes could aid the development of pharmacotherapies to treat alcohol dependence. PMID- 10377524 TI - On ticks and tick-borne diseases. PMID- 10377525 TI - Protective immune mechanisms to ticks and tick-borne diseases of ruminants. AB - A workshop of the European Union (EU) Concerted Action Project on the Integrated Control of Ticks and Tick-borne diseases (CA-ICTTD) recently assessed protective immune mechanisms to ticks and tick-borne diseases. The consensus achieved there is summarized here by Patricia Preston and Frans Jongejan. The current understanding of this field is expanded upon by the accompanying articles, and Poster, in this special issue of Parasitology Today. PMID- 10377526 TI - Immunology of the tick-host interaction and the control of ticks and tick-borne diseases. AB - The first experimental vaccination against ticks was carried out 60 years ago. Since then, progress has been slow, although the recent commercial release of a recombinant vaccine against Boophilus microplus is significant. The nature of naturally acquired protective immunity against ticks is poorly understood, particularly in the important, domesticated ruminant hosts. Characterization of the antigens of naturally acquired immunity remains limited, although more has been achieved with 'concealed' antigens. Crucial questions remain about the true impact of tick-induced immunosuppression and the effect of immunity on the transmission of tick-borne diseases, despite some fascinating and important recent results, as discussed here by Peter Willadsen and Frans Jongejan. PMID- 10377527 TI - Protective immune mechanisms against Theileria parva: evolution of vaccine development strategies. AB - Theileria parva is an intracellular sporozoan parasite that infects and transforms bovine lymphocytes, causing a severe lymphoproliferative disease known as East Coast fever in eastern, central and southern Africa. In this article, Declan McKeever and colleagues summarize the current understanding of immune mechanisms provoked by the parasite with regard to their role in both pathogenesis and protection. In particular, the influence of genomic polymorphism in parasite and host on the development of immunity is discussed, along with the evolution of current vaccine development strategies as a result of immunological research on the disease. PMID- 10377528 TI - Innate and adaptive immune responses co-operate to protect cattle against Theileria annulata. AB - For many years it was assumed that Theileria annulata resembled T. parva, parasitizing lymphocytes and causing lymphoproliferative disease, with the two species being controlled by similar protective immune responses. Patricia Preston et al. here review the evidence that has led to a different view of T. annulata. It is now thought that the schizonts of T. annulata inhabit macrophages and B cells, and that tropical theileriosis is not a lymphoproliferative disease. Both innate and adaptive responses contribute to recovery from infection and resistance to challenge and cytokines produced by infected and uninfected cells influence the outcome of infection. Partial protection has been stimulated recently by defined recombinant antigens; efficacy depended upon the delivery system. PMID- 10377529 TI - Websites of interest PMID- 10377530 TI - Designing blood-stage vaccines against Babesia bovis and B. bigemina. AB - The tick-transmitted apicomplexan parasites Babesia bovis and B. bigemina cause significant disease in cattle in many tropical and temperate areas of the world. These parasites present a challenge for vaccine development, and yet provide a system for studying the pathogenesis, mechanisms of protective immunity and regulation of host immune responses associated with intraerythrocytic protozoan parasites in a non-rodent species. In this article, Wendy Brown and Guy Palmer review strategies for identifying candidate vaccine antigens of B. bovis and B. bigemina and for priming immune responses to evoke strain crossprotective immunity. PMID- 10377531 TI - Molecular basis for vaccine development against the ehrlichial pathogen Anaplasma marginale. AB - Anaplasma marginale is a tick-transmitted ehrlichial pathogen causing severe morbidity and mortality in livestock on six continents. Development of safe effective vaccines would be greatly facilitated by identification of the protective immune mechanisms and by understanding how the pathogen evades immune effectors to establish persistent infection. In this article, Guy Palmer and colleagues review recent progress in identifying how defined epitopes induce protective immunity and the role of antigenic variation in these epitopes as a mechanism of persistence. PMID- 10377532 TI - Immune responses to Cowdria ruminantium infections. AB - Understanding the basis of protective immunity to Cowdria ruminantium will facilitate the development of an effective subunit vaccine against heartwater in ruminants and contribute to a better definition of protective immune mechanisms to obligate intracellular pathogens in general. Until recently, immunological studies of heartwater in ruminants concentrated solely on antibody responses. Since 1995, the mechanisms underlying cell-mediated immunity of heartwater have been analysed. Progress achieved in these areas is discussed here by Philippe Totte and colleagues, with special emphasis on ruminants, the natural hosts of C. ruminantium. PMID- 10377533 TI - Vaccine strategies for Cowdria ruminantium infections and their application to other ehrlichial infections. AB - Suman Mahan and co-authors review the strategies applied to develop improved vaccines for Cowdria ruminantium infections (heartwater). Inactivated vaccines using cell-cultured C. ruminantium organisms combined with an adjuvant are capable of protecting goats, sheep and cattle against lethal C. ruminantium challenge. Immune responses induced with this vaccine, or after recovery from infection, target outer membrane proteins of C. ruminantium, in particular the major antigenic protein 1 (MAP-1). Genetic immunizations with the gene encoding MAP-1 induce protective T helper cell type 1 responses against lethal challenge in a mouse model. Similarly, homologues of MAP-1 in other phylogenetically and antigenically related ehrlichial agents such as Anaplasma marginale and Ehrlichia chaffeensis are also targets of protective responses. Given the antigenic similarities between the related ehrlichial agents, common strategies of vaccine development could be applied against these agents that cause infections of importance in animals and humans. PMID- 10377535 TI - Corrigendum PMID- 10377534 TI - Immune responses to Dermatophilus congolensis infections. AB - Complex mechanisms underly the establishment of dermatophilosis, an exudative and proliferative skin disease of ruminants. This multicomponent system involves the bacterium Dermatophilus congolensis, transmission by various routes including flies, host genetic factors and immunosuppression by Amblyomma variegatum ticks. Here, Nick Ambrose and colleagues summarize recent evidence for an association between A. variegatum and severe chronic dermatophilosis in cattle. Breed-based differences in resistance to dermatophilosis are probably related to immunity to ticks or resistance to the immunosuppressive effects of ticks. Immunity to dermatophilosis might involve non-classic responses mediated by CD1 antigen presentation and gammadelta T cells. Progress towards vaccination is further complicated by strain-specific acquired immunity to D. congolensis. PMID- 10377538 TI - High-frequency brain activity: perception or active memory? PMID- 10377537 TI - Reply. PMID- 10377536 TI - What does semantic dementia reveal about the functional role of the perirhinal cortex? PMID- 10377539 TI - Reply. PMID- 10377540 TI - Multistable phenomena: changing views in perception. AB - Traditional explanations of multistable visual phenomena (e.g. ambiguous figures, perceptual rivalry) suggest that the basis for spontaneous reversals in perception lies in antagonistic connectivity within the visual system. In this review, we suggest an alternative, albeit speculative, explanation for visual multistability - that spontaneous alternations reflect responses to active, programmed events initiated by brain areas that integrate sensory and non-sensory information to coordinate a diversity of behaviors. Much evidence suggests that perceptual reversals are themselves more closely related to the expression of a behavior than to passive sensory responses: (1) they are initiated spontaneously, often voluntarily, and are influenced by subjective variables such as attention and mood; (2) the alternation process is greatly facilitated with practice and compromised by lesions in non-visual cortical areas; (3) the alternation process has temporal dynamics similar to those of spontaneously initiated behaviors; (4) functional imaging reveals that brain areas associated with a variety of cognitive behaviors are specifically activated when vision becomes unstable. In this scheme, reorganizations of activity throughout the visual cortex, concurrent with perceptual reversals, are initiated by higher, largely non-sensory brain centers. Such direct intervention in the processing of the sensory input by brain structures associated with planning and motor programming might serve an important role in perceptual organization, particularly in aspects related to selective attention. PMID- 10377541 TI - Mechanisms of selection for the control of hand action. AB - Most attention research has viewed selection as essentially a perceptual problem, with attentional mechanisms required to protect the senses from overload. Although this might indeed be one of several functions that attention serves, the need for selection also arises when one considers the requirement of actions rather than perception. This review examines recent attempts to determine the role played by selective mechanisms in the control of action. Recent studies looking at reach-to-grasp responses to target objects in the presence of distracting objects within a three-dimensional space are discussed. The manner in which motor aspects of the reach-to-grasp response might be influenced by distractors is also highlighted, rather than merely addressing the perceptual consequences of distractors. The studies reviewed here emphasize several factors highlighting the importance of studying selective processes within three dimensional environments from which attention and action have evolved. PMID- 10377542 TI - Possible stages in the evolution of the language capacity. AB - Much current discussion of the evolution of language has concerned the emergence of a stage in which single vocal or gestural signals were used symbolically. Assuming the existence of such a stage, the present review decomposes the emergence of modern language into nine partially ordered steps, each of which contributes to precision and variety of expression. Bickerton's proposed 'protolanguage' falls somewhere in the middle of this succession. In addition to the by-now accepted evidence from language learning, language disorders, and ape language experiments, modern languages provide evidence of these stages of evolution through the presence of detectable 'fossils' in vocabulary and grammar. PMID- 10377543 TI - Introduction PMID- 10377544 TI - Physical medicine and rehabilitation at the Mount Sinai medical center during the 20th century. AB - The Department of Rehabilitation Medicine at the Mount Sinai Medical Center has a long history, beginning in 1910 with the establishment of the Department of Physical Therapy, headed by Heinrich Wolf, M.D. In 1935, William Bierman, M.D., was appointed director. He was, at that time, one of the leading physicians of physical therapy in the United States, and a prolific researcher, writer and clinician. In 1948, the name of the department was changed to the Department of Physical Medicine, reflecting the newly established specialty of the Board of Physical Medicine. In 1959, Lawrence Wisham, M.D., was appointed chairman, and shortly thereafter the name of the department was changed to Physical Medicine and Rehabilitation. Under Dr. Wisham's leadership, services were provided to inpatients on the acute wards of the hospital, and to outpatients. In 1968, the name of the department was changed to Rehabilitation Medicine. In 1986, Kristjan T. Ragnarsson, M.D., became chairman of the department, and shortly thereafter an inpatient rehabilitation medicine service was established and outpatient services expanded. Since that time, rapid growth has occurred within the department, particularly in delivery of clinical services and research. The role of rehabilitation medicine in the delivery of clinical services to people with temporary or permanent disability is now well established, but efficient and effective delivery must be ensured in the current and future healthcare environment. PMID- 10377545 TI - Geriatrics and rehabilitation medicine: common interests, common goals. AB - Geriatrics and rehabilitation medicine are both fields with roots in antiquity, and they are both objects of renewed interest in modern times. They share a common philosophy: concern for the total needs of the patient. Both fields employ a team approach to patient care. The rehabilitation medicine physician (physiatrist) has an important role in the care of the elderly. Geriatricians and physiatrists should work together for research purposes and to improve the prevention and treatment of illness and disability in the aging population. PMID- 10377546 TI - Physiatric management of mild traumatic brain injury. AB - Mild traumatic brain injury (MTBI) is a common condition, afflicting as many as 1.5 million Americans yearly. Most individuals sustain MTBI as a result of motor vehicle collisions, but it may also occur as a result of falls, physical assault or sporting accidents. Problems related to MTBI include various pain syndromes, cognitive impairments, disorders of affect, cranial nerve dysfunction, and vertigo, arising from injury to the brain, head, or cervical spine. Symptoms are usually transient, although a small percentage of afflicted individuals develop long-lasting problems, often preventing them from leading productive lives. Recognition of these problems as arising from MTBI is difficult due to the frequent lack of abnormal findings on diagnostic tests and failure to identify a history of head trauma. The American Congress of Rehabilitation Medicine has defined MTBI, an important first step in identifying individuals who need treatment. Diagnosis is usually made by directed questions regarding trauma history and careful procurement and interpretation of appropriate tests. Once a diagnosis is made, proper care can be prescribed in order to lead patients toward more productive lives. PMID- 10377547 TI - Quality of life as a construct in health and disability research. AB - Definitional issues that affect the measurement of quality of life (QOL) in health care research are discussed. In reviewing a broad sample of health- and disability-related QOL studies, the authors note several characteristics in which respective approaches to measurement differ: (a) In various measurement tools, QOL has been located either within the insider's (i.e., the person being measured) judgment of the 'goodness' of his or her life or outside this judgment. (b) The insider's and/or outsider's values may hold sway in deciding the elements of life that are relevant to QOL within the measurement process, and in rating the degree of 'goodness' of these life domains. (c) QOL models incorporate domains of items varying in breadth and specificity; and they take either a negative or neutral view of functioning. (d) QOL models vary in their complexity, type of linkage between components, and inclusion (or not) of both the insider's judgment and external predictors of QOL. These distinctions are used by the authors in recommending approaches to QOL measurement suitable for health care research aimed at outcome assessment and description of populations. PMID- 10377548 TI - Acute management of traumatic cervical spinal cord injuries. AB - Successful outcome from a traumatic cervical spinal cord injury (SCI) depends heavily upon the quality of the acute care rendered to the affected individual. In recent years, there have been significant advances in the acute management of SCI. We discuss current management strategies in the areas of prehospital care and transport, emergency room management, surgical considerations and pharmacotherapy. PMID- 10377549 TI - Functional status and its uses in rehabilitation medicine. AB - BACKGROUND: Over the past decade and a half, rehabilitation medicine has developed and implemented standardized measures of functional status. Standardized measures of functional status are important for four reasons: (1) clinicians need them to determine whether interventions produce the expected outcomes; (2) managed care companies use them to decide which rehabilitation services and equipment will be paid for; (3) accreditation bodies such as the Commission on the Accreditation of Rehabilitation Facilities (CARF) require empirical functional status and functional outcome measures; and (4) public policy is moving toward a case-based payment system derived from patient need, and type and severity of impairment. METHODS: Review of the literature. CONCLUSIONS: While researchers, clinicians, managed care, accrediting bodies, and federal regulation have each influenced rehabilitation as conceptualized, measured, and practiced, lack of coordination among these groups has hampered agreement on appropriate tools for functional assessment and outcome. Rehabilitation providers, however, will be increasingly accountable to government regulations and managed care companies. PMID- 10377550 TI - Primary care for people with disabilities. AB - People with disabilities are a unique population. Although there have been great advances in their care, access to reliable and consistent primary health care remains a difficult issue for many of these patients after discharge from medical rehabilitation units. Many of these health care needs are not unique to this patient population, but become compounded or exacerbated in people with disabilities. The effects of physical impairments on these patients' health need to be recognized. Specific attention must be paid to prevent the occurrence of secondary disabilities, which can drastically affect their independence. Ultimately, proper attention to the health care needs of people with disabilities will result in greater independence and improved health among this population. PMID- 10377551 TI - Repetitive strain injury (cumulative trauma disorder): causes and treatment. AB - BACKGROUND: The computerization of the workforce in the last two decades has led to increases in the incidence of repetitive strain injury (RSI). U.S. Workers Compensation claims made by persons disabled in the upper extremities in 1989 were estimated to be $563 million. METHOD: Through an investigation of factors that increase the likelihood of contracting an RSI in various industries, the authors suggest a relation between workplace conditions, lack of education about causes of RSI and improper use of equipment among contemporary workers. RESULTS: A medical term which analyzes the unique history and mechanism of the injury and offers a unique regimen of exercise, education, modalities and some pain management has proven to be the most effective treatment for RSI. Damage to muscle and tendon due to RSI generally cannot be surgically repaired. However, specific nerve-related disorders can be treated with surgery if other more conservative treatments prove ineffective. CONCLUSION: Through foresight and education, industries in which specific factors render workers more likely to contract RSI can begin to take steps to minimize worker susceptibility to the disorder. Those industries will face large losses due to Workers' Compensation claims, disability pay, lawsuits and ultimately lower productivity. PMID- 10377552 TI - Osteoporosis and exercise: a review. AB - BACKGROUND: Osteoporosis is a widespread bone disorder which primarily effects postmenopausal women, with annual costs of six billion dollars. Several studies have looked at the potential value of exercise as an inexpensive and widely available treatment. METHODS: Using Medline, English-language articles published after 1989 in major medical journals and containing the key words 'exercise' and 'osteoporosis' were obtained. Selected prospective studies which included twenty or more subjects were reviewed with special attention to the exercise portion of the protocol. Studies felt to be historically important were also reviewed. RESULTS: Quality of studies, design types and exercise intervention varied greatly. Detailed exercise protocols, high compliance rates and low drop-out rates appeared to favorably effect results. CONCLUSION: As a whole, the literature suggests that exercise induced improvement in bone mineral density in select individuals. Based on the review the author suggests an ideal program. PMID- 10377553 TI - Treatment with an anabolic agent is associated with improvement in respiratory function in persons with tetraplegia: a pilot study. AB - BACKGROUND: Pulmonary complications are a major cause of morbidity and mortality among individuals with cervical spinal cord lesions. Strengthening of the respiratory musculature may reduce these complications. Anabolic steroids have been used to increase muscle mass and improve muscle performance. Oxandrolone, an anabolic steroid, may have beneficial effects on breathing in persons with tetraplegia. METHODS: The effect of one-month treatment with oxandrolone on weight gain and pulmonary function was studied in ten subjects with complete motor tetraplegia. Spirometry, maximal inspiratory and expiratory pressures, and resting self-rating of dyspnea (Borg Scale) were measured at baseline and repeated again at the end of one month of oxandrolone therapy (20 mg/day). Serum lipid profiles and liver function tests were performed before and after treatment. A paired t-test was used to determine pre- and post-treatment differences on the dependent variables. Percent change from baseline was calculated for each variable and tested using a one-sample t-test. RESULTS: On average, the subjects gained 1.4+/-1.5 kg, a 2+/-2% increase in weight (p=0.01). A significant, 9+/-2% improvement was found in the combined measures of spirometry (p<0.005). Maximal inspiratory pressure improved an average of 10+/-7% (p<0.001). Maximal expiratory pressure improved 9+/-13% (non-significant). Subjective self-rating of dyspnea decreased an average of 37+/-28% (p<0.01). CONCLUSIONS: In healthy subjects with tetraplegia, the use of oxandrolone was associated with significant improvements in weight and pulmonary function, and a subjective reduction in breathlessness. Therefore, oxandrolone may be indicated to strengthen respiratory musculature in individuals who have tetraplegia and ventilatory insufficiency aggravated by superimposition of pneumonia or other such conditions. However, long-term use of oxandrolone may not be indicated, due to the adverse complications associated with this class of agents. PMID- 10377554 TI - Psoriasis and Crohn's disease. PMID- 10377555 TI - [High molecular weight fragmentation of DNA and its possible role in genomic rearrangement]. PMID- 10377556 TI - [RNA polymerase II]. PMID- 10377557 TI - [Immunochemical identification of protein products of three new genes from the p13-14 region of human chromosome 11]. PMID- 10377558 TI - [Interaction of an isolated subunit of sigma-70 with oligodeoxyribonucleotides, identical to regions -10 and -35 of the spc promoter]. PMID- 10377559 TI - [Photoaffinity modification of human 80S ribosome using an analog of the mRNA derivative of the hexaribonucleotide pUUUGUU, bearing an arylazide group on the guanine residue]. PMID- 10377560 TI - [Protein environment of the nucleotide U-1061 in the "790 stem-look" region of 18S rRNA in human ribosomal 40S subunit]. PMID- 10377561 TI - [Analysis of two segments of biallelic polymorphism at the tumor necrosis factor in patients with multiple sclerosis from the Russian population: connection with NcoI-PDGF in the first intron of the lymphotoxin alpha gene]. PMID- 10377562 TI - [Structure and function of the Erwinia herbicola speA gene]. PMID- 10377564 TI - [Homologous locus of genomes of Bacillus subtilis and Bacillus stearothermophilus, containing levansucrase and levanase genes]. PMID- 10377563 TI - [Antirestriction activity of metalloregulatory proteins ArsR and MerR]. PMID- 10377565 TI - [Polymorphism of the angiotensinogen gene and genetic predisposition to diabetic nephropathy in type I diabetes mellitus]. PMID- 10377566 TI - [Intracellular endonuclease from Serratia marcescens. I. Spatial structure of the protein and crystalline state at a resolution of 1.7 A]. PMID- 10377567 TI - [The role of small-scale movement in the protein globule in intensifying the electron interactions of photoactive groups]. PMID- 10377569 TI - [Conformational analysis of DNA in complexes with E. coli RNA polymerase. Modeling and experimental data]. PMID- 10377568 TI - [Stability of the structure of KRP (kinase related protein) protein]. PMID- 10377570 TI - [Refinement of helix boundaries in alpha-helical globular proteins] ]. PMID- 10377571 TI - [Analysis of lymphoma-specific transcripts in B-cell non-Hodgkin's lymphoma patients, infected by the human immunodeficiency virus type 1]. PMID- 10377572 TI - [Changes in the composition of transcription factor AP-1 in rat embryonal fibroblast cells, transformed by E1A and Ha-Ras oncogenes]. PMID- 10377573 TI - [DNA fragments from regions of the inter-disk polytene chromosomes of Drosophila melanogaster bind the nuclear matrix in vitro]. PMID- 10377574 TI - [Expression of alpha-fetoprotein and hepatocyte nuclear factors in rat hepatoma clones: effect of retinoic acid]. PMID- 10377575 TI - [Transformation of immortalized rat embryonic fibroblasts by the E7 gene of human type 16 papillomavirus]. PMID- 10377576 TI - [The proposed role of the ceruloplasmin receptor and Cu2+-transferring Menkes ATPase in copper metabolism in mammals]. PMID- 10377577 TI - [Structure-activity organization of the cowpox strain GRI-90 viral genome. II. Comparative analysis of the structure of the left species-specific region of the orthopoxvirus genome]. PMID- 10377579 TI - [Microsatellite (ttgc)n, specific for the intergenic spacer of human and chimpanzee rDNA: use for studying the structural variations of the prepromoter region of rDNA]. PMID- 10377578 TI - [Structure-activity organization of the cowpox strain GRI-90 viral genome. III. Functional characteristics of the left species-specific region of the genome]. PMID- 10377580 TI - [Secondary structure of hairpin 17 of the lower multicellular animal Rhopalura ophiocomae (Mesozoa: Orthonectida) as an example of "punctuated equilibrium" in the evolution of 18S ribosomal RNA]. PMID- 10377582 TI - [The role of prenatal diagnosis on decreased incidence of congenital defects in the pediatric population of the Czech Republic 1990-1996]. AB - The author presents data on the prevalence of birth defects in the Czech Republic during the period from 1990-1996. The paper contains also data on results of prenatal diagnosis of birth defects in the Czech Republic during the same period and indicates achievements in secondary prevention of abnormal phenotypes, which contributed in a major way to the reduced incidence of some types of inborn defects in the population. PMID- 10377581 TI - [Diagnosis of Martin-Bell syndrome based on an analysis of the structural functional changes in the 5'-untranslated region of the FMR1 gene]. PMID- 10377583 TI - [Ultrasound imaging of the lower urinary tract in post-menopausal women with urinary stress or combined type of incontinence before and after intravaginal administration of estriol]. AB - The finding that climacteric symptoms are caused mainly by a decline of the oestrogen level and that their development can practically always be prevented by long-term local or general oestrogen treatment was a great asset to the treatment of this problem. Oestriol, a less effective natural oestrogen, has a favourable effect on urogenital tissues without stimulating the endometrium [2]. The objective of the present investigation was to analyze ultrasonographic parameters of the lower urinary tract in women after the menopause with the stress or mixed type of urinary incontinence before and after two-month local oestriol treatment (Ovestin). The trial comprised 40 women with confirmed stress (GSI) or the mixed type of urinary incontinence. The group with GSI comprised 124 patients and the group suffering from the mixed type of incontinence comprised 26 women. The type of incontinence was assessed by urogynaecological examination. This was followed by transperineal and introital ultrasound examination of patients in a supine position by means of an Acuson 128 XP10 apparatus using a convex probe with a frequency of 5 MHz and a vaginal probe with a frequency of 7.0 MHz. Assessment of the position and mobility of the urethrovesical junction was implemented by the transperineal route using a convex probe and filling the bladder with 300 ml. After urination followed assessment of the urethral sphincter by the introital route in a vertical plane whereby the authors followed the anterior and posterior surface of the rhabdosphincter, and in a horizontal plane its left and right surface (10). The authors assessed also in both planes the maximal thickness of the sphincter. In the vertical plane and in a proximal position in relation to the urethra they evaluated the vascular supply qunatitatively (minimal-1 to very abundant-4) and also the arterial flow-the pulsatile index PI was investigated as well as the resistance index RI. In the vertical plane 1 cm from the urethrovesical junction the authors assessed the thickness of the urethral mucosa; at the same level they evaluated the thickness of the urinary bladder wall; the anterior wall, the vertex and the area of the trigone. They assessed also the thickness of pelvic floor muscles. The assessments were made before and after two-month intravaginal oestriol administration (Ovestin crm)-two weeks 0.5 mg/day and then 0.5 mg twice a week. After treatment no statistically significant differences in thickness and areas of the urethral sphincter were found nor in the thickness of the pelvic floor muscles before and after oestriol administration. Statistically significant differences were recorded in the mobility of the urethrovesical junction and there was a significant increase in the thickness of the urethral mucosa and a more abundant vascularization was recorded during the quantitative evaluation and evaluation of PI. In women with the mixed type of incontinence after oestrogen treatment a decline in the thickness of the urinary bladder was found. Ultrasound examination of the lower urinary tract before and after oestriol treatment (Ovestin crm) is a useful supplement of common examination methods and it confirms its favourable therapeutic effect when administered by the intravaginal route. PMID- 10377585 TI - [Cerclage--yes or no]. PMID- 10377584 TI - [Does prenatal administration of thyrotropin releasing hormone contribute to premature delivery?]. PMID- 10377586 TI - [Indications for and risks of invasive methods of prenatal diagnosis--who indicates and who carries the risk?]. PMID- 10377587 TI - [Fetomaternal alloimmune thrombocytopenia--possibilities of diagnosis and exchange transfusion]. PMID- 10377588 TI - [Pitfalls in obstetrical peridural analgesia and anesthesia]. PMID- 10377589 TI - [Ropivacaine--a new local anesthetic for obstetrical peridural analgesia]. PMID- 10377590 TI - [Epididymal and testicular sperm in the in vitro fertilization program: the origin of sperm does not affect the results of intracytoplasmic sperm injection]. PMID- 10377591 TI - [Therapy and therapeutic results in carcinoma of the cervix at the Masaryk Oncologic Institute in Brno 1987-1992]. PMID- 10377592 TI - [The 5th edition of the TNM classification--malignant gynecologic tumors]. PMID- 10377593 TI - [The 5th edition of the TNM classification--malignant tumors of the breast]. PMID- 10377594 TI - [Diagnosis and therapy of early cervical pregnancy]. PMID- 10377595 TI - [Is determination of flow in the ductus venosus and umbilical vein of clinical importance?]. PMID- 10377596 TI - [The role of laparoscopy in gynecologic oncology]. PMID- 10377597 TI - [Angiogenesis in the human reproductive system]. PMID- 10377598 TI - [Intervening variables and vitamins in pregnancy]. PMID- 10377599 TI - [The basis of chronic ischemic reversible left ventricular dysfunction (hibernating myocardium)]. AB - The term "Hibernating myocardium" refers to the presence of a chronic left ventricular dysfunction that could be partially or completely improved after revascularization. Formerly, the myocardial hibernation was considered to be an adaptive response to a sustained reduction of the resting myocardial blood flow. Recently, several studies with positron emission tomography have revealed a normal or almost normal resting myocardial blood flow. The reduced coronary flow reserve of the hibernating myocardium brings about multiple episodes of demand induced ischemia and may evoce recurrent stunning of the myocardium. It is the basis of structural changes in the myocardium and the contractile dysfunction. Cellular degeneration rather than adaptation prevails in the hibernating myocardium. The longer the process continues, the smaller is the chance for a complete structural and functional recovery after the reperfusion. Thus the early revascularization becomes important for the patients with hibernating myocardium. PMID- 10377600 TI - [Reassessment of the role of aluminum in the development of Alzheimer's disease]. AB - The pathophysiology of Alzheimer's disease (AD) is related to the alterations in neurotransmission, beta-amyloid production, plaque formation and cytoskeletal abnormalities. The question of aluminium relevance to the etiology of AD cannot yet be adequately answered. Aluminium is currently regarded as the putative risk factor for the disease. Our paper shows that some of pathologic changes are not raised by aluminium alone, but by the aluminofluoride complexes. These complexes may act as the initial signal stimulating impairment of homeostasis, degeneration and death of the cells. By influencing energy metabolism these complexes can accelerate the aging and impair the functions of the nervous system. In respect to the etiology of AD, the long term action of aluminofluoride complexes may represent a serious and powerful risk factor for the development of AD. PMID- 10377601 TI - [Cerebral blood flow measurement]. AB - A cerebral blood flow measurement is an important part of studying various brain pathologies, particularly brain ischemia. The methods of measurement are still improving, although the physical substance of many of them, like Fick's or Doppler's principles, outlasts further. A change in quality in last several years came with the use of isotopes and a computer technique. New options in imaging of circulatory changes with the help of magnetic resonance technology seem to be particularly perspective. If there were no economical obstacles, this method, which is able to disclose non-invasively a brain perfusion disorder in the very beginning, would be certainly immediately applied in the broad clinical practice. PMID- 10377602 TI - The divergence in the conception of Pavlov and Bayliss-Starling concerning the function of the nervous system. AB - The history of the discoveries of the mechanisms of the pancreatic secretion is a fine example of the conflict of two schools and the concurrence of two concepts: The nerve control (Pavlov and his co-workers) and the humoral control (Bayliss and Starling). Only the synthesis of both conceptions brought about the most accurate model of the control mechanisms. PMID- 10377603 TI - [Melatonin and its wide-spectrum effects: use of melatonin in the treatment of tumors]. AB - One of the major physiological roles of melatonin is its synchronizative influence on circadian rhythmicity and the induction of seasonal responses to changes in day length. In seasonally reproductive animals like the hamster, melatonin has been established as antigonadotrophic on the gonadal axis. Melatonin has been used successfully to treat jet lag and some circadian-based sleep disorders. It has the immunomodulatory role, which can be supported by the existence of specific binding sites in lymphoid cells, thymus and spleen. Not negligible is the antioxidative effect of melatonin and the ability to scavenge some reactive forms of oxygen. Melatonin has potentially important influence on the neoplastic growth and direct and indirect oncostatic effect in some forms of neoplasia. The beneficial influence of melatonin alone or its combination with immunotherapy, radiotherapy or chemotherapy in many clinical studies in patients with tumors was demonstrated. PMID- 10377604 TI - Waianae Diet Program: long-term follow-up. AB - A long-term follow-up was conducted on 82 participants from prior programs based on ad libitum feeding of a traditional Hawaiian diet. Follow-up period ranged from 12 months to 90 months and averaged 33.67 months. An average weight loss of 15.1 pounds was maintained over 7.5 years of follow-up (p < 0.0005) even when stratified over two year intervals, suggesting that this type of program may be an effective long-term weight loss intervention. PMID- 10377605 TI - The dietary treatment of inflammatory arthritis: case reports and review of the literature. AB - Two patients with seropositive inflammatory arthropathies who experienced clinical improvement on the Waianae diet are presented. The scientific literature validates the usefulness of fasting in the control of joint inflammation. Elimination diets are variably successful. Fasting followed by a vegetarian diet can produce a sustained positive response measured clinically and by laboratory variables of inflammation; the efficacy of such an approach appears to hinge on the alteration of fecal flora. Swaying the balance of dietary fats in favor of the omega 3 and omega 6 fatty acids has an antiinflammatory effect, but does not appear to correct the basic immunologic processes involved in the development of the arthropathies. Practical guidelines for the application of this information are offered. PMID- 10377606 TI - IPS Empress inlays and onlays after four years--a clinical study. AB - OBJECTIVE: Ceramic inlays are used as esthetic alternatives to amalgam and other metallic materials for the restoration of badly damaged teeth. However, only limited clinical data are available regarding adhesive inlays and onlays with proximal margins located in dentine. In a prospective, controlled clinical study, the performance of IPS Empress inlays and onlays with cuspal replacements and margins below the amelocemental junction was examined. MATERIALS AND METHODS: Ninety-six IPS Empress fillings were placed in 34 patients by six clinicians. The restorations were luted with four different composite systems. The dentin bonding system Syntac Classic was used in addition to the acid-etch-technique. At baseline and after 6 months, one, two and four years after placement the restorations were assessed by two calibrated investigators using modified USPHS codes and criteria. A representative sample of the restorations was investigated by scanning electron microscopy to evaluate wear. RESULTS: Seven of the 96 restorations investigated had to be replaced (failure rate 7%; Kaplan-Meier). Four inlays had suffered cohesive bulk fractures and three teeth required endodontic treatment. After four years in clinical service, significant deterioration (Friedman 2-way Anova; p < 0.05) was found to have occurred in the marginal adaptation of the remaining restorations. Seventy-nine percent of the surviving restorations exhibited marginal deficiencies, independent of the luting composite. Neither the absence of enamel margins, nor cuspal replacement significantly affected the adhesion or marginal quality of the restorations. CONCLUSION: After four years, extensive IPS Empress inlays and onlays bonded with the dentin bonding system Syntac Classic were found to have a 7% failure rate with 79% of the remaining restorations having marginal deficiencies. PMID- 10377607 TI - The effect of saliva on enamel and dentine erosion. AB - The present study aims to assess the ability of saliva, both in vitro and in situ, to prevent surface mineral loss from enamel and dentine when exposed to an erosive challenge. Fifteen groups of four varnished thin tooth sections were stored in saliva collected from individuals taking part in the in situ study and a further eight groups, each containing four sections, were stored in deionised water. In vitro, sections were stored in saliva or water for 14 days. In addition, fifteen subjects each wore an appliance with four varnished sections. Appliances with sections were worn for 14 days. All sections were exposed to 25 ml of erosion solution for 5 min twice daily. Microradiography and image analysis of the recovered sections demonstrated significant protection of surface mineral loss from enamel and dentine by saliva in vitro and in situ compared with deionised water (p < 0.05). Significantly less mineral loss (p < 0.05) was observed for enamel and dentine stored in situ compared with storage in saliva in vitro. Generalised linear modelling demonstrated both the subject and protocol had significant effect on mineral loss. A weak positive correlation (r = 0.64) was noted when in situ and in vitro mineral loss from enamel were compared, demonstrating greater reactivity of the in vitro enamel specimens to the erosion challenge. The dentine data did not show any linear correlation. Saliva protected against mineral loss by erosion and, for enamel, in vitro results demonstrated a weak positive correlation with in situ results. PMID- 10377608 TI - Development and evaluation of a low erosive blackcurrant juice drink. 2. Comparison with a conventional blackcurrant juice drink and orange juice. AB - OBJECTIVE: A previous study demonstrated that an experimental low pH blackcurrant juice drink with calcium was markedly less erosive to enamel both in situ and in vitro than orange juice. Further development of the experimental blackcurrant juice drink formula has occurred and the aims of the present studies were two fold. Firstly, to confirm the low erosivity of the modified formulation and secondly, to provide more comparative data with other acidic fruit drink products. METHOD: The study was a single centre, single blind, randomised placebo controlled 4 cell crossover design involving 12 healthy volunteers. The test drinks were orange juice, water, experimental blackcurrant juice drink with calcium and a proprietary blackcurrant juice drink. Enamel samples were retained in situ at the mid palatal regions using intraoral appliances and exposed to 250 ml volumes of the drinks 4 times per day during 15 working days. Measurements of enamel loss were made by profilometry. The same method was modelled in vitro. RESULTS: By day 15 the mean losses of enamel in situ for orange juice, water, experimental blackcurrant juice drink with calcium and blackcurrant juice drink were 1.70, 0.05, 0.44 and 2.75 microns respectively. At all bar one measurement, the loss of enamel was significantly greater for all fruit drinks compared to water. Losses caused by the blackcurrant juice drink with calcium were significantly less than the other two fruit drinks at all time points. Losses of enamel by day 15 in the study in vitro were 13.02, 0.00, 1.78 and 39.02 microns respectively. The blackcurrant juice drink with calcium was not significantly different from water at days 3 and 6, otherwise all pairwise comparisons for differences between the 4 drinks at each time period were significant. CONCLUSIONS: The methodologies in situ and in vitro again appear to correlate in ranking the order of erosivity of drinks. The data particularly from the study in situ allude to the very low comparative erosivity of the further modified experimental blackcurrant juice drink with calcium and support the further development of such drinks for public consumption. PMID- 10377609 TI - Development and evaluation of a low erosive blackcurrant juice drink. 3. Final drink and concentrate, formulae comparisons in situ and overview of the concept. AB - OBJECTIVE: Two previous studies demonstrated that a blackcurrant juice drink with added calcium produced little erosion of enamel in vitro and in situ by comparison with other low pH fruit drinks. The primary aim of this study was to demonstrate that the final formulation drink and concentrate were of similar low erosivity. Secondary aims were to provide more data on the erosivity of other fruit drink concentrates and whether erosion was influenced by anterior and posterior palate siting of enamel specimens. METHOD: The study was a single centre, single blind, randomised placebo controlled 5 cell crossover design involving 15 volunteers. The test drinks were blackcurrant juice/calcium concentrate, blackcurrant juice/calcium drink, proprietary apple & blackcurrant juice concentrate, proprietary orange drink concentrate and water. Four enamel samples were retained in situ, 2 anterior palate and 2 mid/posterior palate, on upper removable acrylic appliances. Drinks were 250 ml volumes consumed 4 times per day during 15 working days. Concentrates were diluted 50 ml in 200 ml water. Measurements of enamel loss were made on one anterior and one posterior sample on days 2, 5, 10 and 15 by profilometry. RESULTS: One subject approached the 20 microns erosion limit by day 10 on the orange drink and was withdrawn from that cell. Differences in mean erosion between anterior and posterior sites were variable and small. By day 15 the mean losses of enamel averaged over anterior and posterior were blackcurrant/calcium concentrate 0.28 micron, blackcurrant/calcium drink 0.35 micron, apple & blackcurrant concentrate 2.04 microns. orange concentrate 8.29 microns and water 0.08 micron. Except at day 15 for the blackcurrant/calcium drink the erosion by the blackcurrant/calcium formulations was not significantly different from water at any time point. Erosion by the apple & blackcurrant and orange concentrate drinks was highly significantly greater than the blackcurrant/calcium drinks at all but the 2 day time point for the apple & blackcurrant concentrate drink compared to the blackcurrant/calcium drink. CONCLUSIONS: All data thus far indicate that dependant on tooth site susceptibility and the specific drink, the consumption of standard low pH fruit drinks could result in 1 mm loss of enamel in periods ranging from as little as 2 years to 20 years. Similar erosion by the low pH blackcurrant drinks with added calcium would take in excess of 100 years. PMID- 10377610 TI - An in vitro investigation of a poly(vinyl phosphonic acid) based cement with four conventional glass-ionomer cements. Part 1: Flexural strength and fluoride release. AB - OBJECTIVE: To investigate the flexural strength and fluoride release of four conventional glass-ionomer cements: Ketac-Molar (KM), HiFi (HF), Vivaglass Fil (VF), Ketac-Fil (KF) and a newly developed glass polyphosphonate cement, Diamond Carve (DC). METHOD: Disc specimens (10 mm diameter, 1 mm thick) were prepared and mould stored at 37 degrees C. After one hour, the specimens were removed from their mould and immersed in 20 ml of deionised water until required for testing. Biaxial flexural strength was determined at 1 hour and at 1, 7, 30 and 90 days after the start of mixing. Measurements of fluoride release from the specimens were carried out at 2 hours and at 1, 3, 7, 14, 30, 60 and 90 days after the start of mixing using a fluoride ion selective electrode. The results were analysed using ANOVA and student 't' tests. RESULTS: All the materials displayed different flexural strength patterns. KM and DC became stronger whilst KF and VF plateaued in strength with time. HF peaked in strength and then became weaker. At 90 days, the mean flexural strengths in decreasing order was as follows: KM > or = VF > or = DC > or = HF > KF. An initial fast rate of fluoride release followed by a slower but steady release of fluoride was observed in each of the materials. The mean cumulative fluoride release in decreasing order was as follows: VF > KF > or = HF > DC > KM. VF released significantly higher level and KM significantly lower level of fluoride than the other materials. CONCLUSIONS: The acid used to form the cement could not be used to predict changes in cement strength behaviour with respect to time. DC increased in strength with time and its flexural strength at 90 days was comparable to that of HF and VF. The cumulative and rate of fluoride release varied for the materials. DC had a low fluoride release consistent with a fast setting material with good early resistance to water. PMID- 10377611 TI - An in vitro investigation of a poly (vinyl phosphonic acid) based cement with four conventional glass-ionomer cements Part 2: Maturation in relation to surface hardness. AB - OBJECTIVE: To investigate the surface hardness of four conventional glass-ionomer cements: Ketac-Molar (KM), HiFi (HF), Vivaglass Fil (VF), Ketac-Fil (KF) and a newly developed glass polyphosphonate based cement, Diamond Carve (DC) at different maturation times in water and to investigate the effects of early water exposure on their surface hardness. METHOD: Disc specimens (10 mm diameter, 1 mm thick) were prepared and mould stored at 37 degrees C. The effect of different maturation times (15, 30 and 60 min) and storage in water over 24 h after those maturation times on surface hardness was determined using a microindentor with a Vickers diamond indentor. The results of the surface hardness tests were analysed using Mann-Whitney non-parameteric statistics (p < or = 0.05). The working (WT) and setting (ST) times of the cements were also measured using a modified Wilson oscillating rheometer. RESULTS: All the materials became harder after 24 h immersion in water. HF, VF and DC showed initial sensitivity to a short maturation time, but only HF was adversely affected by early moisture exposure. KF and KM were least sensitive to short maturation time or early water exposure. DC had the shortest and HF the longest WT and ST. HF and VF had a high WT:ST ratio of 1:7 and 1:9, respectively. CONCLUSIONS: A short maturation time (of 15 min) and early exposure to water did not adversely affect the surface hardness of KF, KM, VF and DC. DC, based on poly(vinyl phosphonic acid), had the shortest WT and ST. Poly(acrylic acid) based HF and VF had a long ST in relation to their WT. PMID- 10377612 TI - Flexural and thermal cycling of resins for veneering removable overlay dentures. AB - OBJECTIVES: This investigation compared the effect of flexural and thermal cycling upon resins, bonded via adhesive primers to cobalt-chromium alloy to identify appropriate materials for veneering overlay dentures which may flex in function. METHODS: The resins investigated were an acrylic resin, a crown and bridge resin, a urethane dimethacrylate and a hybrid composite resin systems. The first three were bonded via an adhesive primer/opaque system based upon 4 methacryloxyethyl trimellitate anhydride (4-META) and the last via one based on 10-methacryloxyldecyl dihydrogen phosphate (MDP). Cast alloy beams 30 x 4 x 0.8 mm with 2 mm resin veneers were stored in H2O for three days or three months, and subjected to 1000 thermo-cycles between 4 and 60 degrees C, and/or 5000 flexural cycles to a displacement of 0.1 mm. Three-point bend testing was carried out in an Instron 4505 UTM. The increase in the flexural moduli of the cast beams, due to the addition of the resin spines, together with the displacements and loads at yield were recorded. RESULTS: The adhesive resin systems varied in their abilities to withstand conditioning stresses, this appeared to reflect the rigidity of the resin component as well as the performance of the adhesive bond. CONCLUSIONS: While the composite/MDP system maintained the highest bond strength throughout the conditioning, its low displacement at yield indicates that it may be more suitable for rigid areas of a removable partial denture. Acrylic resin, with its high load and displacement at yield, make it the material of choice for veneering the more flexible saddle areas of partial dentures. PMID- 10377613 TI - Failure of resin-modified glass-ionomers subjected to shear loading. AB - The mechanism of bond failure of resin-modified glass-ionomers is unknown. This study examined the failure on shear loading at the dentine interface of these materials. Twenty-five teeth (embedded in acrylic blocks) were sectioned longitudinally to expose a flat dentine surface. Cylinders of materials were made by injecting into a tube placed on the dentine of each section surface. The materials used were Fuji Cap II and Fuji II LC (GC Corp., Japan), Vitremer (3M Dental Products, USA), Photac-Fil (original) and Photac-Fil* (new) (ESPE Dental AG, Germany). After a week, a fluorescent dye was placed in the pulp chamber of each tooth and left for 3 h. The specimens were sectioned through the cylinders before both halves were tested in shear. The failure was observed using a confocal microscope, with video rate images (stored) digitally. The shear load at failure and locus of failure were recorded. All specimens had intact interfaces before testing, except the original Photac-Fil specimens which dislodged from their tooth surfaces even before testing, while being mounted on the device. An amorphous zone or absorption layer was noted at the dentine interface of 60% of Fuji II LC, 22% of Vitremer and all of the Photac-Fil* (new) specimens, but not in Fuji Cap II. Failure was cohesive in Fuji II LC, adhesive in Vitremer, cohesive/adhesive in Photac-Fil* (new) and cohesive in Fuji Cap II. In specimens with the absorption layer present, the failure was at the material/absorption layer interface, leaving it behind on the dentine surface. The mean stresses at failure (MPa) and standard deviations were 5.60, 2.46 (Fuji II LC); 4.82, 0.99 (Vitremer); 4.97, 2.10 MPa (Photac-Fil*); and 3.48, 1.06 (Fuji Cap II). All data were normally distributed as tested by the Shapiro-Francia test. One-way analysis of variance using exact inferential statistics indicated no significant difference between the mean failure stress for all the systems, p = 0.08. The mechanism of failure of resin-modified glass-ionomer materials to shear loading at the dentine interface varies between products. In materials in which the absorption layer is present, it appears to play an important role in mediating the bond of the glass-ionomer to dentine. PMID- 10377614 TI - Comparison of polymerization contraction stresses between self- and light-curing composites. AB - OBJECTIVES: The objective of this study was to examine the distributions and the magnitudes of the internal stresses in self- and a light-curing composite restorations resulting from polymerization shrinkage. METHODS: Butt-joint box shaped cavities (5.0 x 2.0 mm2, 2.0 mm in depth) prepared in composite molds were filled with either a self- or light-curing transparent resin composite. The restorations were cross-sectioned perpendicular to the longitudinal axes of the cavities and observed using polarizing microscopes. The principal stresses in the restorations, normal and shear stresses at the cavity wall were evaluated by photoelastic analysis. RESULTS: The distributions of the principal stresses and the stresses generated at the cavity wall in both the self- and the light-curing composite restorations were similar. The maximum stress generated at the cavity wall in the light-curing composite restorations was twice as large as that seen in the self-curing restorations. CONCLUSIONS: The results of this study indicated that the difference in the magnitude of the internal stresses between self- and light-curing composites was not related to the distribution of the stresses. The velocity of polymerization appeared to be the most important factor contributing to the magnitude of the internal stresses generated in the composite restorations in this study. PMID- 10377615 TI - Effect of dentin primers containing N-methylolacrylamide or N methylolmethacrylamide on dentin pretreatment. AB - The effect of experimental dentin primers containing N-methylolacrylamide (MEAA) or N-methylolmethacrylamide (MEMA) on the bonding of three commercial light-cured resin composite systems [Restorative Z 100 (Scotchbond), Palfique Estelite (Macbond) and Photo Clearfil A (Clearfil Photobond)] to etched dentin was investigated. Water solutions of 35% hydroxyethyl methacrylate (HEMA). 50% MEAA or 30% MEMA were used as dentin primers. The dentin etched with 10% phosphoric acid solution was pretreated with dentin primers for 30 s. The resin composite systems were applied in a Teflon tube positioned onto pretreated dentin surfaces. After water immersion for 1 day, the shear bond strengths were measured. The thicknesses of hybrid layers at dentin-resin interfaces treated with 6 mol/l HCl and 1% NaOCl were measured by scanning electron microscopy. The dentin primer pretreatment increased the bond strengths of all resin composite systems. For Macbond and Clearfil Photobond, the bond strengths (14.2-26.5 MPa) with MEAA and MEMA were higher than those (10.5 and 17.8 MPa) with HEMA. All hybrid layer thicknesses were 1-1.5 microns after HCl immersion. The hybrid layers after NaOCl immersion become narrower. The main fracture pattern of specimens exhibiting high bond strengths (more than 14.2 MPa) was dentin cohesive fracture after bond test. For Macbond and Clearfil Photobond, the layers of specimens pretreated with MEAA and MEMA were clearly thicker than those pretreated with HEMA after NaOCl immersion. MEAA and MEMA solutions were more effective in improving the bond strength of Macbond and Clearfil Photobond to etched dentin than was HEMA. Macbond and Clearfil Photobond created good hybrid dentin layers which could resist NaOCl-attack when MEAA and MEMA solutions were used as dentin primers. PMID- 10377616 TI - The time is now. PMID- 10377617 TI - Education. Where have all the future nurses gone? PMID- 10377618 TI - Inquiry, insights, and history. Nursing leadership of uncontrolled pain. PMID- 10377619 TI - International affairs. Caring for older people in the next millennium. PMID- 10377620 TI - Legal and ethical issues. Ethics content in nursing education. PMID- 10377621 TI - Why nurses must actively participate in the debate on assisted suicide: a symposium. AB - This article argues that nursing must undertake further careful study of assisted suicide. Nursing must determine the relationship of assisted suicide to its core values as an important condition for making beneficent, patient-centered clinical decisions as well as for participating in discussions that establish public policies and professional practice standards. PMID- 10377622 TI - The moral appeal of assisted suicide in end-of-life decisions. AB - This article identifies conceptual distinctions, e.g., killing, letting die, causing, refraining from causing, and moral principles, eg, autonomy, beneficence, utility, and their implications for the debate on assisted suicide. PMID- 10377623 TI - Legal decisions and public opinion informing the debate on assisted suicide. AB - This article examines the evolution of state and federal legislation and court opinions in the 1990s concerning treatment abatement and assisted suicide. The recent Supreme Court decision on assisted suicide is summarized, and its rejection of a recognized constitutional right to assisted suicide is explored. Additionally, surveys of the opinions of nurses, physicians, and the public regarding the permissibility of assisted suicide are evaluated. The contradictions between public opinion and some federal and state legislation are highlighted and discussed. PMID- 10377624 TI - Assisted suicide and nursing: possibly compatible? AB - This article argues that the American Nurses Association's (ANA) prohibition of nurse-assisted suicide is misguided. The ANA's reasons for this policy do not provide the necessary conceptual or empirical support for the prohibition. In fact, arguments appear to lead to support for nurse-assisted suicide: (1) because the claim that death is always harmful may be false, the obligation to "do no harm" does not necessarily preclude assisted suicide (AS); (2) currently we have no evidence that AS would erode public trust in nurses; (3) AS may be compatible with the professional integrity of nursing, particularly the commitments to respecting autonomy, promoting patient welfare, and providing compassionate care; (4) nursing's participation would constrain, rather than contribute to, the potential for abuse to vulnerable patient populations; and (5) the professional has a responsibility to either embrace the public's increasing support of aid in dying or determine why AS is morally indefensible and educate the public. PMID- 10377626 TI - Outcomes assessment: implications for nursing education. AB - In response to public criticisms about the outcomes of higher education, colleges and universities have implemented comprehensive assessment programs. In nursing education, outcomes assessment has become a criteria for accreditation. Yet, currently no guidelines exist describing "best practices" in nursing education outcomes assessment. Experts in educational assessment provide helpful guidelines for nurse educators as they establish comprehensive assessment plans. The authors provide a review of the literature in the field of assessment and offer strategies for developing successful assessment programs in nursing education. PMID- 10377625 TI - Changing the present legal prohibitions on assisted suicide is a bad idea. AB - This article argues that although there may exist morally valid reasons to justify assisted suicide in some cases, establishing state or national policies legalizing assisted suicide is morally unjustified. Five arguments are examined that support the position against legalization: consequentialist arguments, arguments about who could legitimately choose assisted suicide and when it could be chosen, the incompatability of assisted suicide policy and authentic self determination; the probable lack of efficacy of assisted suicide in a managed care environment, and the effects of assisted suicide on the profession and practice of nursing. The article ends with a discussion of alternatives to assisted suicide legislation that might accomplish in large part what is sought by such legislation. PMID- 10377627 TI - Women's ways of knowing in nursing and critical thinking. AB - The purpose of this longitudinal qualitative study was (1) to extend the work of Belenky, Clinchy, Goldberger, and Tarule by interviewing female university nursing students to determine their "way of knowing" according to the Women's Ways of Knowing (WWK) schema and (2) to determine what relationship this way of knowing might have with critical thinking when accumulating a specific body of knowledge such as nursing. Interviews were conducted with 21 sophomore nursing students. Fourteen were reinterviewed their junior year, and 10 were interviewed or participated in a focus group their senior year. The procedural knowledge categories of separate and connected knowing became the focus of data analysis through the constant comparative method. Procedures for connected knowing were illuminated. Connected knowing was found to be congruent with nursing and the ways these women wanted to be as nurses. Separate knowing was found to be incongruent with nursing except for critical thinking purposes. Contrary to the notion that critical thinking is principled rather than procedural, procedural knowing, according to WWK, became the principle on which these women based their nursing actions, moving them to constructed knowers and caring, critical thinkers as they experienced nursing education. PMID- 10377628 TI - School of nursing closure: an example of congruence with Sutton's model of organizational death. AB - Literature concerning the closing of nursing schools is scarce. When faced with the task of closure, educational administrators are without guides because most nurses have had no experiences in implementing what can be termed "organizational death." The purpose of this article is to describe Sutton's Process Model of Organizational Death and its congruence with the closure of one nursing school. Process model knowledge and application may assist administrators, faculty, and all those connected with a nursing program in positively working through a school closure. PMID- 10377629 TI - Nursing attire: indicators of professionalism? AB - The purpose of this qualitative research was to explore the effect that current nursing attire has on the image of the nursing profession. A number of nurses and a nonnurse were interviewed to determine how attire affected their perception of today's nurses. The two research questions were as follows: (1) is the changing dress of nurses projecting a negative image to the public? and (2) What components of a nurse's apparel indicate professionalism? Content analysis was performed on transcriptions from the tape-recorded responses of a purposeful sample of health care workers: 12 registered nurses, 1 bachelor of nursing student, and 1 layperson. The responses for the first research question were not directly addressed by the participants. However, one overall theme emerged, which was labeled "I can't tell you what it is, but I know it when I see it." The main theme that emerged for the second research question was labeled "total package," with role identification and competency being related themes. As a result of this research, nursing administrators and other health care professionals could gain an understanding of the importance of nursing attire as an indicator of nursing professionalism. Future research needs to examine the same research questions with health care consumers in a variety of acute and community-based health care settings. PMID- 10377630 TI - Should nurses practice therapeutic touch? Should nursing schools teach therapeutic touch? PMID- 10377631 TI - Should nurses practice therapeutic touch? Should nursing schools teach therapeutic touch? PMID- 10377632 TI - Capricious nonsense. PMID- 10377633 TI - Dualistic approach. PMID- 10377634 TI - Nonmetal crowns. PMID- 10377636 TI - Antibiotics not always necessary. PMID- 10377635 TI - More to it than 'fix my tooth'. PMID- 10377638 TI - Quantitation of vital bleaching by computer analysis of photographic images. AB - BACKGROUND: The authors investigated the use of computer processing of photographic images to monitor changes in tooth brightness after nightguard vital bleaching, or NGVB. METHODS: Photographs of shade guides and clinical cases (patients' teeth) were taken on 35-millimeter film with electronic flash illumination and processed commercially. A slide scanner was used to digitize images as red, green and blue, or RGB, files, with constant brightness, contrast and linearity settings; the images were then analyzed with commercial software. Relevant image components (that is, teeth or shade guide tabs) were separated, and histograms of various numerical color descriptors were generated for each image component. RESULTS: Analysis of shade tab images showed that the mean pixel intensity for the RGB blue channel, or MPIb, was the most satisfactory brightness descriptor, with clear sequential MPIb increments from lighter to darker shades in each series of colors (A through D) and close correlation with the manufacturer's brightness scale (r = .83). Mathematical analysis of MPIb data for shade tabs in the same image yielded a brightness index that was reproducible and correlated well with the manufacturer's brightness scale. Sequential measurements of this index in three subjects whose teeth were bleached with carbamide peroxide for 14 days correlated well with assessments made by visual shade guide comparisons. CONCLUSIONS: The authors conclude that computer analysis of digitized photographic images with internal color controls provides an index of tooth brightness that is reproducible from image to image. CLINICAL IMPLICATIONS: A brightness index derived from computer analysis of digitized photographic images may be useful for monitoring the effectiveness of NGVB. PMID- 10377637 TI - Pain after periodontal scaling and root planing. AB - BACKGROUND: Although periodontal scaling and root planing, or SRP, is one of the most common procedures used in dental practice, there is little information available about the degree of postprocedural pain associated with it. The authors undertook this study to document the intensity and duration of pain after SRP with a view toward helping practitioners and their patients manage postprocedural discomfort. METHODS: Using the Heft-Parker self-assessment pain scale, 52 adults with moderate periodontitis evaluated their pain before and after SRP conducted with local anesthetic. RESULTS: After SRP, 28 percent of all patients reported faint-to-weak pain, 18 percent experienced weak-to-mild pain, 28 percent experienced mild-to-moderate pain, 8 percent had moderate-to-strong pain and 8 percent reported strong-to-intense pain. The average time to onset of maximum pain was approximately three hours after SRP, and the average duration of mild or greater pain was about six hours. Upon awakening the morning after SRP, subjects found that pain had returned to pre-SRP levels. Overall, 23 percent of all patients reported self-medicating with analgesics to relieve postprocedural pain. Women self-medicated earlier (P < .05) and more often than men (43 percent vs. 10 percent; P < .05). CONCLUSIONS: Patients experienced significant duration and magnitude of pain after SRP. This pain peaked between two and eight hours after SRP, lasted about six hours, and returned to pre-SRP levels by the morning after the procedure. Almost 25 percent of all patients self-medicated to relieve pain after SRP, and women took analgesic medication earlier and more often than men. CLINICAL IMPLICATIONS: Practitioners should consider using appropriate analgesic drugs to alleviate mild-to-moderate pain after SRP. On the basis of this study, it would appear that an analgesic that has a peak effect two to eight hours after the completion of SRP would be the most appropriate medication. Moreover, it is unlikely that analgesic medication would be needed by most patients beyond the day on which SRP was performed. PMID- 10377639 TI - Evaluating the effects of fluoride-releasing dental materials on adjacent interproximal caries. AB - BACKGROUND: The authors examined several restorative materials to evaluate their ability to inhibit demineralization and enhance remineralization of incipient carious lesions on the interproximal enamel of teeth adjacent to those restored with the materials. METHODS: Twenty-one subjects in need of a crown on a mandibular molar and a Class II inlay on an adjacent tooth took part in this six phase study. Artificial enamel lesions were created and positioned within the interproximal portion of a crown. Lesions were photographed with polarized light microscopy and characterized before and after 30-day intraoral exposures. Each phase included the placement of a new section in the crown model and a new Class II inlay restorative material in the adjacent tooth. RESULTS: Results demonstrated that nonfluoridated resin composite, fluoridated resin composite and resin-modified glass ionomer restorative materials, when placed in subjects who brushed with a fluoridated dentifrice, demonstrated significantly (P < .05) less enamel demineralization than the nonfluoridated resin composite control placed in subjects who brushed with a nonfluoridated dentifrice. The resin-modified glass ionomer cement, however, even when brushed with a nonfluoridated dentifrice, exhibited significantly (P < .05) less demineralization than the nonfluoridated resin composite control brushed with a nonfluoridated dentifrice. CONCLUSIONS: Resin-modified glass ionomer cement appears to significantly inhibit demineralization of interproximal enamel of teeth adjacent to those restored with the material. CLINICAL IMPLICATIONS: Resin-modified glass ionomer cement restorations can enhance prevention of enamel demineralization on adjacent teeth. PMID- 10377640 TI - When patients become cyanotic: acquired methemoglobinemia. AB - BACKGROUND: The authors conducted literature review to create a heightened awareness of the potential for developing toxic methemoglobinemia from local anesthetics. Methemoglobin normally is present in the blood at levels less than 1 percent. Levels may become toxic as hemoglobin is oxidized to methemoglobin after local anesthetics such as benzocaine and prilocaine are administered. TYPES OF STUDIES REVIEWED: The authors searched the medical and pharmaceutical industry literature. They found and reviewed case studies of incidences of methemoglobinemia that resulted from local anesthetic overdoses. RESULTS: Cases of local anesthetic-induced methemoglobinemia in dental practice are under recognized and rare. Reported cases of prilocaine-induced methemoglobinemia have resulted in recent changes in some prilocaine literature. These changes include maximum recommended doses for patients of various weights. CLINICAL IMPLICATIONS: Dentists should identify patients who are at increased risk of developing methemoglobinemia before administering local anesthetics. They also should follow new recommended dosing guidelines for prilocaine and be aware of symptoms of this adverse reaction. PMID- 10377641 TI - Diagnosis and treatment of cutaneous facial sinus tracts of dental origin. AB - BACKGROUND: Cutaneous draining sinus tracts of dental origin often are a diagnostic challenge. A delay in correctly diagnosing these types of lesions can result in ineffective and inappropriate treatment. CASE DESCRIPTION: The authors present five cases of facial lesions that were initially misdiagnosed as lesions of nonodontogenic origin. The correct diagnosis in each case was cutaneous sinus tract secondary to pulpal necrosis and suppurative apical periodontitis. All facial sinus tracts resolved after the patients received nonsurgical root canal therapy. CLINICAL IMPLICATIONS: As patients with cutaneous facial sinus tracts of dental origin often do not have obvious dental symptoms, possible dental etiology may be overlooked. Early correct diagnosis and treatment of these lesions can help prevent unnecessary and ineffective antibiotic therapy or surgical treatment. PMID- 10377642 TI - The automated external cardiac defibrillator: lifesaving device for medical emergencies. AB - BACKGROUND: More than 350,000 adult Americans die each year of sudden cardiac arrest, or SCA. The event is unpredictable and can occur in patients with no history of cardiac disease or cardiac symptoms. Drugs and cardiopulmonary resuscitation, or CPR, save only a small percentage of victims. The necessary response is rapid application of electrical shock, and the chances of success are reduced 10 percent for every minute of delay. TYPES OF STUDIES REVIEWED: The author reviewed the literature on resuscitation of people who have undergone SCA, and examined the emerging technology of automated external defibrillators, or AEDs, for correcting cardiac ventricular fibrillation. Included is a review of the controversies surrounding AED waveforms and energy levels. RESULTS: Automated cardiac defibrillators are becoming readily available because of improved technology and decreasing prices. AEDs are now commonly found in commercial aircraft, gambling casinos, sports arenas and public buildings, and will soon become as readily available as fire extinguishers. The use of AEDs is being taught in standard CPR courses. CLINICAL IMPLICATIONS: AEDs are being installed in more public locations, including some dental offices. As costs decrease and availability increases, there is significant potential use for AEDs in managing SCAs in dental offices. PMID- 10377643 TI - A simplified approach to isolating a single tooth before endodontic therapy. PMID- 10377644 TI - Online continuing dental education. AB - BACKGROUND: An increasing number of continuing dental education, or CDE, courses are available on the Internet. The authors conducted this study to determine general characteristics of online CDE course offerings. METHODS: The authors found online CDE courses through Internet search engines and dental indexes. They recorded each course's Web page address, title, topic, length, credit hours and cost. Then they classified course providers, categorized topics, compared course length with credit hours and calculated cost per credit hour. RESULTS: The authors located 157 online CDE courses offered by 32 providers. The courses covered a wide range of topics, and most were five screens long or shorter. Credit hours per screen ranged from 0.05 to two, and cost per credit hour ranged from no charge to $25. CONCLUSIONS: Online CDE courses are hard to locate, making this material accessible only to people who are well-versed in retrieving information on the Internet. The brevity of most courses may make them appropriate for incremental study at the dental practitioner's convenience. Guidelines to correlate credit hours with course length should be developed. PRACTICE IMPLICATIONS: Online CDE courses may become an important tool to help practitioners keep current. Several issues, however, need to be addressed before online CDE can reach its full potential. PMID- 10377645 TI - The cost-effectiveness of a new chlorhexidine delivery system in the treatment of adult periodontitis. AB - BACKGROUND: Periodontal treatment is costly. The authors assessed the potential economic impact of a new periodontal chemotherapeutic, testing the hypothesis that its adjunctive use would result in reduced periodontal surgical needs. METHODS: An economic model estimated treatment needs following two clinical trials of the adjunctive use of a chlorhexidine, or CHX, -containing chip compared with scaling and root planing, or SRP, alone. Needs were based on periodontal status at nine months and a probabilistic algorithm; costs were assigned on the basis of a national dental survey and an average wholesale price of the CHX chip. RESULTS: The base case model projected significantly more maintenance procedures and significantly fewer periodontal surgical procedures for patients treated with SRP and the CHX chip compared with patients who were treated with SRP alone (54.4 percent vs. 46.4 percent, P = .014; 29.2 percent vs. 35.5 percent, P = .015, respectively). Average total costs of care for patients treated with SRP and CHX chip were $737 +/- $244 compared with $734 +/- $239 for patients treated with SRP alone. Sensitivity analyses to account for variations in practice patterns did not appreciably alter the results. When data were analyzed after only three or six months of treatment, the significant differences in treatment needs disappeared. CONCLUSIONS: The CHX chip is a new, apparently cost-effective treatment option for non-surgical periodontal therapy. Adjunctive use of the CHX chip could reduce periodontal surgical needs significantly at little or no additional cost. CLINICAL IMPLICATIONS: Results suggest that incorporating the CHX chip into routine practice requires a new algorithm for management of periodontal disease. To obtain full clinical benefit, treatment needs to be continued for nine months. PMID- 10377646 TI - Learning, immunology and allergic responses. PMID- 10377648 TI - Supreme Court rules in ERISA pre-emption case. PMID- 10377647 TI - Adjunctive periodontal therapy. PMID- 10377649 TI - [New methods for the measurement of the burden of disease in Sweden]. PMID- 10377650 TI - [The value of gastrointestinal tumor surgery should be scrutinized. The therapeutic results are not always correctly interpreted]. PMID- 10377651 TI - [Intravenous immunoglobulin therapy is more and more common. The increasing tempo can force a prioritization of patients, scientists are warning]. PMID- 10377652 TI - [Gamma-globulin shortage threatens a patient group]. PMID- 10377653 TI - [Hardly any research funds to women]. PMID- 10377654 TI - [HPV-tests as suitable complement to cytological screening]. PMID- 10377655 TI - [Screening for diseases is futile as a preventive method in connection with tobacco]. PMID- 10377657 TI - [Confusing proposal for revisions in the Helsinki declaration]. PMID- 10377656 TI - [Difficult decisions on estrogen treatment of women with coronary diseases]. PMID- 10377658 TI - [Excessive neuropsychiatric diagnosis is not beneficial]. PMID- 10377659 TI - [The AD-drops can be replaced by D-drops]. AB - Since 1932, when vitamin A and D supplementation, in the form of cod liver oil, was introduced in Sweden, rickets has been a rare diagnosis among Swedish infants. In 1978, the National Board of Health and Welfare issued recommendations of daily supplementation with 300 micrograms (1000 IU) of vitamin A and 10 micrograms (400 IU) of vitamin D. This has recently been under review by the Paediatric Committee on Nutrition and Health, of the Swedish Paediatric Association and the National Food Administration, who concluded that there is no reason to retain vitamin A supplementation, but that vitamin D supplementation should continue to be recommended at the same daily dose (400 IU). PMID- 10377660 TI - [Recommendations for prevention of iron deficiency. Delay cow's milk intake as a beverage to infants until 10-12 months of age!]. AB - Breast-feeding is to be encouraged during the first six months of life. Iron deficiency is extremely rare in exclusively breast-fed infants during this period. Any cow-milk based formula used should be iron-fortified. During the second half of infancy, the iron content of weaning foods is important in preventing iron deficiency. Indeed, owing to the low iron content of dairy products, it is hard to compose a weaning diet sufficiently rich in iron to meet the demands of rapidly growing infants, if it is to include substantial amounts of cow milk, sour milk or yoghurt. Accordingly, the Paediatric Committee on Nutrition and Health, of the Swedish Paediatric Association and the National Food Administration, recommend delaying the introduction of cow's milk and cow-milk products until the infant is 10-12 months of age. Until then, breast-feeding, and the use of iron-fortified formula or gruel with modified protein and sodium content are encouraged; iron-fortified porridges of softer consistency can be prepared to circumvent the need of extra fluids, or porridge can be served with breast milk or iron-fortified formula; small amounts of milk may be used for cooking purposes. PMID- 10377662 TI - [Order and comfort in Konradsberg--the first modern psychiatric hospital in Sweden]. PMID- 10377661 TI - [A study of experience and consequences of diabetes. Increased psychosocial support in the care of diabetes is needed]. AB - Having diabetes affects life in many ways. The achievement of well-functioning self-care is highly dependent on the patient's ability to come to terms with the disease as an integral part of life, an integration which in turn is crucially dependent on the patient's emotional experience of the disease and trustful relationships with the health care professionals involved. Thus, the caregiver's professional expertise must include knowledge of the medical, psychological and social aspects of the disease. The article consists in a review of the results and conclusions of a population study designed to examine insulin-dependent diabetics' experience of living with their disease, their use of, and attitudes to, diabetes care, and the social and psychosocial impact of the disease on their lifestyle. The principal determinants of the patients' relationship both to the disease itself and to diabetes care appeared to be gender and the presence or absence of chronic complications. Another finding was the need of increased psychosocial support expressed by many patients, which suggests medical social workers to be important members of the diabetes care team. PMID- 10377663 TI - [Successful prevention of tractor accidents in Sweden]. AB - A decrease in the incidence of tractor roll-over fatalities, from 15/100,000 farm tractors for the period 1957(60 to 0.1/100,000 for the period 1986(90, was associated with a corresponding increase in the proportion of farm tractors equipped with roll-over protection structures (ROPS) from 6 to 93 per cent. Thus, the Swedish approach to the prevention of serious injuries due to tractor roll over, entailing the compulsory equipment of new tractors with ROPS from July 1st 1959, may be said to have been successful. No other country seems to have introduced such effective safety regulations for the prevention of these injuries. PMID- 10377664 TI - [Young children are endangered when close to a tractor. Keep children away from tractors; make the tractors safer!]. PMID- 10377665 TI - [Urinary incontinence in men--a neglected problem? A quarter of all 80-year old men suffer of urinary leakage]. AB - Urinary incontinence is an increasing health problem in industrialised countries, and has been shown to affect more the eight per cent of the adult population. Incontinence accounted for about two per cent of Swedish health care costs in 1990, and the costs are rising. Although the diagnosis and treatment of female incontinence has attracted more attention since the condition is more prevalent among women, the impact on quality of life is no less among men. As a variety of causes and underlying diseases may result in involuntary urine leakage, careful evaluation is mandatory before treatment is initiated. The article consists in a review of common diagnoses and treatment options in cases of male incontinence. PMID- 10377666 TI - [Over 900 workers in different health professions have received qualification for stroke care]. PMID- 10377667 TI - [Lament of a travel medicine specialist: "Why don't they do what we tell them?" Perhaps the tourists want to hold on to their image of paradise]. PMID- 10377668 TI - [Rossini--an extravagance in tones and calories]. PMID- 10377669 TI - [Contagion, contagious and contagiousness--a linguistic reflection. Infectious is worse than infective, but the worse is full of infection]. PMID- 10377670 TI - [Rules of Tove Jansson serve as guidance for physicians]. PMID- 10377671 TI - [Treatment with cytostatic agents should be considered in colorectal cancer. It improves prognosis and quality of life]. PMID- 10377672 TI - [Rehabilitation after stroke is beneficial]. PMID- 10377673 TI - [Afraid of labor, afraid of pain--individual management is needed. "Planned delivery" is an alternative to section]. PMID- 10377674 TI - [Is society going to finance projects of drug companies?]. PMID- 10377675 TI - [Homocysteine as a deficiency marker. Attitudes and values]. PMID- 10377676 TI - [Concentrate on geriatrics]. PMID- 10377677 TI - [Meta-analysis of therapeutic trials. Risk of underestimating specific effects]. PMID- 10377679 TI - ["Good old advice" (about breast feeding) must be--good!]. PMID- 10377678 TI - [Depressed adolescents say no to professional help. Prevention--illusion or hope of the future?]. PMID- 10377680 TI - [Incorrect information about suggested changes in the Helsinki declaration]. PMID- 10377681 TI - [Interleukin-1 and nitric oxide involved in the pathogenesis of diabetes)]. AB - Insulin-dependent diabetes mellitus (IDDM) is not only a common metabolic disorder in industrialised countries, but its incidence is still increasing, especially in Scandinavia. The article consists in a review of evidence implicating nitric oxide (NO) and the cytokine, interleukin-1 (IL-1), in the pathogenesis of IDDM. Cytotoxic effects of IL-1 and NO, generated through autoimmune reactions associated with insulitis and impairing the function of insulin-producing pancreatic beta-cells in IDDM, are discussed, as are possible pharmacological strategies for blocking this toxicity. Compounds capable of blocking IL-1 cell surface receptors and NO synthesis may prove beneficial in protecting beta-cells from autoimmune assault in IDDM. If IL-1 causes beta-cell dysfunction and destruction through NO synthesis in IDDM, several pathways in the IL-1-NO system are attractive potential targets for drugs protecting beta-cells against these effects, thus providing a means of intervening in the pathogenesis of IDDM. PMID- 10377682 TI - [For whom is training after stroke most beneficial? Selection method exists and should be used]. AB - In the light of recent Swedish health care cost cuts, this review addresses the need of limiting stroke rehabilitation to those it will benefit most. In many prognostic stroke studies, end-points have been adopted that vary as to time and place--e.g., duration of hospitalisation or discharge to a nursing home. Even modest improvement in ability may significantly enhance quality of life, but is not necessarily associated with beneficial effects on lifestyle or the length of hospital stay. Accordingly, we need to use reliable tests with more realistic end points. The use of the Functional Independence Measure (FIM) and the Katz ADL scale as prognostic instruments is discussed. PMID- 10377683 TI - [Anterior cruciate ligament injuries--still an enormous challenge. Life style related surgery likely to be used more widely]. AB - Anterior cruciate ligament (ACL) injuries still constitute the greatest single problem in orthopaedic sports medicine. The natural history is not well known. Conservative treatment is appropriate in cases where there are no instability problems, and where activity modifications are acceptable to the patient. However, there is an increased risk of meniscus and cartilage injuries, and thus of arthrosis. The indication for surgery depends on the level of activity, and on whether instability symptoms are present. Arthroscopic surgery of the ACL seems to yield good short-term results in 80-90 per cent of cases. Out-patient ACL surgery is becoming increasingly common. Rehabilitation is often vigorous, with early mobilisation and weight-bearing. Functional range-of-motion and closed kinetic chain exercises predominate. Successful surgical outcome often allows former levels of activity to be resumed. There is still uncertainty as to whether surgery reduces the risk of arthrosis. The nature of the original trauma, and continued top level sports activity may be factors of importance. Further research is needed to elucidate the natural history, and determine the long-term effects of modern ACL surgery. PMID- 10377684 TI - [Recommendations are not changed--let infants sleep in supine position. Surgical specialists' contribution to population studies would be of value]. PMID- 10377685 TI - [Asymmetric skull in children? Make a correct early diagnosis!]. PMID- 10377686 TI - [Spiritual needs of severely ill patients are neglected in health care]. AB - Recent decades have witnessed improvements in palliative care in terms of the physical, social and psychological needs of patients with advanced disease. Although interest in established religions has waned in the western world during the same period, many patients nevertheless have spiritual needs and in this respect there is room for improvement in palliative care. The article consists in discussion of the spiritual needs of patients with advanced disease, and how they can be met by care givers. PMID- 10377687 TI - [Quality assurance of blood gas analysis--a medical risk zone]. AB - Blood gas analysis is fundamental to all intensive care. Although speed is essential for adequate treatment, prognosis and even survival of the patient, precision and consistency of results are equally important prerequisites for correct clinical decision making. Point-of-care testing (POCT) has become one of the predominant fields of blood gas analysis. Although modern technology and instruments are user friendly, special training and continuous updating of staff education are of paramount importance to the reliability of results. The article consists in an outline of quality requirements and discussion of appropriate procedures for assessing and maintaining quality in blood gas analysis. PMID- 10377688 TI - [Infant food is not only a question of nutritional physiology. Doubtful to advise against milk with porridge]. PMID- 10377689 TI - [The proposal of the EU concerning experimental animals is a threat to biomedical research. The aim should be fewer animals used per project and not a general reduction]. PMID- 10377690 TI - [Far too few schoolchildren use protective helmets when bicycling. Review of the literature and questionnaires as basis for promotion of increased use of the helmets]. PMID- 10377691 TI - [Living in poverty undermined health of the graphic artist David Tagstrom from Falun]. PMID- 10377692 TI - [The role of "Lijecnicki vjesnik" in the training of Croatian physicians]. PMID- 10377693 TI - [Current approaches in surgical treatment of ulcerative colitis (reports of our results)]. AB - The results of surgical treatment of ulcerative colitis in 19 patients in the period from 1989 to 1997 are presented. Of surgical techniques, in 17 patients a pouch was formed, namely, in six patients W pouch, in four patients S pouch, and in seven patients J pouch, while in two patients the pouch was not formed due to the incontinence of anal sphincter. In these patients the stool derivation was achieved by permanent terminal ileostomy. Fifteen patients were operated in elective program, while in four patients emergency surgery was indicated. In emergency surgery the first step was subtotal colectomy, with ileostomy and sigmoidostomy. In further treatment we introduced corticosteroid clyster in the opening of sigmoidostoma, with good results. The quality of life of persons with the pouch is higher compared to patients with permanent ileostomy. In patients who require surgical treatment of ulcerative colitis, the operation of forming an ileal pouch and anal anastomosis should be done. PMID- 10377694 TI - [Epidemiology of Crohn's disease in the Rijeka-Istra region]. AB - The incidence and prevalence of Crohn's disease in Rijeka and Istra, the western part of Croatia with 575,000 inhabitants, were investigated for the period 1973 1994. A total of 197 patients were diagnosed. This gave an annual incidence of 0.34/10(5) in 1973 and 3.47/10(5) in 1994. These data suggest that incidence of Crohn's disease in our region is similar to that reported in northern Europe. The most frequent age groups affected were 15-25 and 50-60 years old. 54% of the patients were younger than 30 years. The small bowel was involved in 49.7% of the patients, the large bowel in 23.3%, and both small and large bowel in 25.8%. At the time of diagnosis 85.8% of patients had abdominal pain, 84.2% diarrhea, 71.6% significant weight loss. All forms of intestinal and extraintestinal complications were observed. Surgery was required in 25.5% patients. No patient developed a malignancy of the gastrointestinal tract. One patient had a cancer of uterus and one a cancer of vulva. The diagnosis was based on clinical features in combination with endoscopic and radiologic findings, and in most patients it was also supplemented by histologic examination. The mean interval from the symptom onset to diagnosis was 1.9 years. The positive family history among relatives was found. In Crohn's disease there was no dose-response relation between smoking and disease. The relationship should be further explored because of the potential for other therapeutic approaches in this disease. PMID- 10377695 TI - [Effect of improved surgical anesthesiologic factors on the results of treatment of aneurysms of the abdominal aorta]. AB - The purpose of this study was to compare the recent results of the abdominal aortic aneurysm (AAA) surgery, with the results for the patients operated ten years ago, and to identify the factors influencing the operative results. Two groups of patients were selected using a retrospective case series study. First group of patients (A) consisted of 32 cases operated in the period between 1984 and 1986, and the second group (B) consisted of those operated ten years later (from 1994 to 1996). Mortality rate, presence of risk factors, demographic data, and operative factors were analyzed and compared between the groups. The mortality rate decreased from 10.7% in period A, to 5.3% in period B for elective operations, and from 75% to 44.4% for emergency operations. The prevalence of straight graft technique in period B showed statistically significant difference compared to period A (p < 0.005). Besides, the operating time and the amount of the transfused blood between the groups differed significantly (p < 0.05). On the basis of this data, we concluded that the modifications in the operative technique, as well as the increased number of operations within the second period led to the decreased mortality rate in patients operated for AAA. PMID- 10377696 TI - [Prognosis for life expectancy using the inductive learning method]. AB - The aim of this paper was to find out the possibility of life expectancy achievement (LEA) prognosis in open population by using epidemiological data, to improve it and to determine the differences between regions. We were using inductive learning software tool, ASSISTANT Professional, based on modified Quinlan's inductive learning method. Data from an epidemiological study of oil/fat consuming influence on diabetes incidence in different regions in Croatia, 1887 examinees, have been used. In spite of limited number of attributes that were available, an improvement in prognosis of LEA has been made by selecting the proper attributes and by changing the limiting values of attributes (values that change the meaning of attributes). We reached the absolute accuracy of 75.54%. Even after further pruning of decision tree that lowered this value, Assistant showed 13% better result than those of random selected outcome. Differences between regions were also established that could not been explained with attributes that were used. Expanding the list of attributes and analysis of their influence in particular region can make further improvement in prognosis of LEA. PMID- 10377698 TI - [Rickettsiales--modern findings]. AB - The term rickettsiae has as a rule encompassed the intracellular bacteria. Molecular studies brought new data to rickettsial taxonomy. Many new data on rickettsia have been accumulated over recent years, and a comparison of the newly discovered diseases with previously known rickettsioses, as well as with their agents is of interest. The number of "new" agents that have been discovered in the last few years is remarkable. This review discusses the current knowledge of bacterial species that historically belonged to the order Rickettsiales, as well as the role of these agents as human and animal pathogens. PMID- 10377697 TI - [Primary adrenocortical micronodular dysplasia]. AB - Two girls (11 and 13 years old) with Cushing's syndrome due to primary adrenocortical micronodular dysplasia (PAMD) are presented. High plasma cortisol concentrations, elevated urinary free cortisol and 17-ketogenic steroids excretion, in addition to low or normal plasma adrenocorticotropic hormone (ACTH) levels pointed towards independent adrenal cortisol hypersecretion. In both girls bilateral adrenalectomy was performed, followed by replacement therapy with glucocorticoids and mineralocorticoids. Pathohistological findings of otherwise enlarged adrenal glands, showed characteristic small nodules measuring 1-2 mm, composed of cells resembling those of zona fasciculata, with abundant, clear cytoplasm. Our younger patient fulfilled the criteria of "Carney complex", because beside PAMD she has had the lentigines. PMID- 10377700 TI - [A new minimally invasive neurosurgery method in the treatment of refractory epilepsy: vagus nerve stimulator implantation]. PMID- 10377699 TI - [Pathophysiology of hemodynamic shock (circulatory failure). Part 2]. AB - In the second part of the review authors stress the importance of extent and duration of tissue shock hypoenergosis and body reactive capacity for the clinical outcome of the syndrome. Functional restitution, decrease of functional organ capacity, permanent absence of certain organs' function and death, represent a possible clinical status caused by and developed during the shock syndrome. Progressive pathologic alteration of tissue function and structure correlates well with the degree of tissue hypoenergosis. A short detailed description of the tissue alterations is outlined in the paper. The shock syndrome very often consists of parallel pathogenic processes which therefore can be classified as a complex pathogenic forms of the shock. A list of clinical disorders which develop due to a complex shock pathogenesis, are outlined in the paper. Tentative relative contribution of individual pathogenic processes are estimated for various diseases. Clinical symptoms and signs, as well as laboratory parameters give a valuable information which points to the level of shock development and reversibility. Correlation of clinical parameters and pathophysiologic processes are outlined. Simple predictive rules are re-discussed in the scope of underlying pathophysiology. In addition, a related hemodynamic disorders are shortly discussed in the paper. PMID- 10377701 TI - [Citations as a reflection of international impact of scientific work and a measure of individual influence on the development of science in Croatia]. PMID- 10377702 TI - [Citations as an international reflection of scientific work and measure of individual effect on the development of science in Croatia]. PMID- 10377703 TI - [Citations as a reflection of international scientific work and measure of individual effect on the development of science in Croatia]. PMID- 10377704 TI - A medical readiness model of health assessment or well-being in first-increment air combat command medical personnel. AB - We used a medical readiness model of health assessment based on the Neuman systems model, a comparative-descriptive design, to assess the health or state of well-being of Air Combat Command medical personnel. Group I consisted of 636 personnel actively participating in medical readiness training, and group II consisted of 127 personnel assigned to a medical treatment facility that recently returned from overseas deployment. In group I, statistically significant differences in developmental, psychological, and sociocultural elements of health varied according to military rank, mobility status, or previous overseas deployment experience. The spiritual element of health differed statistically in both groups according to military rank. The uncertainties of mobility status, lack of previous deployment experience, and contrasts in military rank increased stress. Recommendations include realistic mobility training concentrating on essential job performance elements, leadership providing clear and open channels of communication, and dealing effectively with the emotional impact of humanitarian assistance missions. PMID- 10377705 TI - The Tripler LE3AN Program: a two-year follow-up report. AB - This paper provides a review of 2-year follow-up data on the Tripler Army Medical Center LE3AN Program. The LE3AN Program (emphasizing healthy Lifestyles, reasonable Exercise, realistic Expectations, Emotions, and Attitudes, and Nutrition) provides active duty service members a treatment strategy that involves a reasonable low-intensity exercise regimen, behavior modification, intensive nutritional counseling healthy meal planing, relapse prevention strategies, cognitive coping strategies, and healthy lifestyle principals to lose weight and maintain weight loss. Based on the 2-year data and additional clinical findings, we expand upon earlier preliminary reports. The 2-year follow-up data suggest that the program is a safe and efficacious treatment program. PMID- 10377706 TI - A conceptual overview of a proactive health psychology service: the Tripler Health Psychology Model. AB - The military patient population, the demanding environment in which medical services are provided, and the rigors of the operational environment create a unique challenge for service members as well as military health care providers. Within the military medical system, the subspecialty of clinical health psychology may provide patient care and consultation interventions necessary to meet the demands of the unique Army medical and military communities. As funding and other resources decrease, military health psychologists can provide high quality care to difficult-to-manage patients while increasing outcome efficacy and decreasing costs to the hospital. This paper provides a definition of clinical health psychology and a description of its unique interventions and applications and how these unique skills augment medical services. Moreover, we offer a conceptual model for an innovative health psychology program that will assist other military treatment facilities in designing programs to increase outcome efficacy and concurrently reduce costs and utilization of services. PMID- 10377707 TI - Psychiatric illness and the workplace: perspectives for occupational medicine in the military. AB - In inpatient psychiatric wards and outpatient mental health clinics throughout the military, psychiatrists and other mental health professionals are often faced with patients suffering from emotional distress attributed to occupational stress. There has been scant research into how the routine military work environment affects the mental health status of military employees. This paper provides a review of the occupational medicine literature on the relationship between the work environment and employee mental health. There is a growing recognition that stress resulting from the workplace can provoke psychiatric illness, but the research is limited at this time. The data existing on the work force in general are examined, and the relationship of these findings to the military work environment is discussed. This review suggests that a comprehensive examination of the relationship between the military work environment and the mental health of military employees is needed. By gathering these data, interventions can be planned to mitigate the effect of stress caused by the military work environment on the mental health of its members. PMID- 10377708 TI - Knowledge and use of birth control methods in active duty Army enlisted medical trainees. AB - This study was designed to determine the familiarity of medical advanced individual training (AIT) students with current methods of birth control and to evaluate the accessibility of these methods. A survey was distributed to 578 medical specialist AIT students assigned to Fort Sam Houston, Texas, for training. Results obtained show a lack of knowledge concerning the newer forms of contraceptives available. This study also indicates that barriers may exist that limit a soldier's ability to acquire prescription forms of contraception while in training. PMID- 10377709 TI - The false-negative fraction: a statistical method to measure the efficacy of cervical smear screening laboratories. AB - The false-negative fraction (FNF) is emerging as a statistical parameter that may be used to evaluate the efficacy of Papanicolaou smear screening laboratories. Our objectives for this paper are to acquaint non-laboratorians with this important measurement and to measure the FNF of the Air Force Cyto-center (AFCC) at the Armed Forces Institute of Pathology in Washington, DC. The FNF is defined as estimated false negatives divided by (true positives plus estimated false negatives). Most often, the number generated is multiplied by 100 and expressed as a percent. We have determined the FNF of the AFCC to be 3.7%. This value compares favorably with most others reported in the medical literature. PMID- 10377710 TI - Occupational back disability in U.S. Army personnel. AB - Musculoskeletal disorders represent a prevalent source of outpatient visits, lost work time, hospitalization, and disability in the military. Recent research has identified patterns among military occupations, gender, and musculoskeletal disability. Although back disorders accounted for a high percentage of all cases, little is known about the relationship between job type and disability in soldiers. The present study analyzed 41,750 disability cases to determine (1) prevalence of work-related back disability diagnoses, (2) specific jobs associated with greater risk of back disability, and (3) association among gender, job type, and disability. The results indicate that (1) lumbosacral strain and intervertebral disc syndrome represent the most prevalent diagnoses for back disability, (2) certain occupations were associated with higher back disability risk, and (3) specific jobs were identified in which females experienced higher rates of back disability than males. The nature of these high risk jobs, and recent research on work disability factors in U.S. Army soldiers, suggest that a combination of ergonomic and individual/organizational psychosocial factors may play a role in the development, exacerbation, and maintenance of work disability. Future research that identifies specific job factors contributing to increased back disability risk should assist in the development of empirically based work site prevention programs to improve musculoskeletal health and readiness. PMID- 10377711 TI - Postmortem percutaneous core biopsy of the liver. AB - Unexpected deaths related to significant hepatopathology are encountered daily by forensic pathologists. After investigation, a decision will be made regarding whether or not to perform an autopsy. In considering the options available to assess the degree of hepatopathology, a study was undertaken to evaluate the potential role of percutaneous core biopsy of the liver at the time of postmortem examination. Postmortem percutaneous hepatic core biopsy was attempted at the time of external examination in 28 nonconsecutive cases examined at the Office of the Chief Medical Examiner, State of Maryland; hepatic tissue was obtained in 26 cases. In 21 of these cases, hepatic tissue was obtained during subsequent partial or complete autopsy and submitted for histologic correlation. There was complete histologic correlation of tissue obtained via percutaneous biopsy with hepatic tissue obtained by open biopsy in 18 of 20 cases (86%). Significant hepatopathology was identified by core biopsy in 5 of 9 cases (56%) with a history of ethanol abuse and in 8 of 19 cases (42%) with a negative ethanol history. The sensitivity of this technique was 82% and the specificity was 100%. This study has shown that core biopsy of the liver can provide information related to the cause of death in cases with and without a history of alcohol consumption. In cases in which a complete autopsy is deferred because of familial religious objections, infectious disease, or time/budgetary constraints, this modality can be used to obtain diagnostic tissue. PMID- 10377712 TI - The patient flow of wounded marines within a multi-echelon system of care. AB - Hospitalization data were extracted for Marines wounded in Vietnam from 1965 to 1969 to examine the echelon flow of treatment care for different types of injuries. The inter-echelon movement of each patient who was hospitalized at an echelon II or III facility was tracked until the treatment was completed or until the patient was moved to a facility in the continental United States. Results showed that approximately half of the admissions to echelon II or III facilities had no further treatment recorded at a higher echelon of care. Almost one-fourth of the patients required treatment at an echelon IV facility, and more than one third were admitted to an echelon V facility. PMID- 10377713 TI - Fitness, performance, and risk of injury in British Army officer cadets. AB - The objectives of this study were to investigate the effectiveness of the Commissioning Course (CC) to develop and maintain standards of fitness, to assess capability to perform military tasks, and to determine the relationship between fitness and risk of injury in 106 British Army officer cadets (OCdts). Aerobic fitness, muscular strength, endurance, and body composition were regularly assessed. Performance on four representative military tasks was measured at the end of the CC. All injuries in term 1 were documented. Over the CC, changes in fitness were generally modest and equivocal. Aerobic fitness improved by approximately 10% (p < 0.01), strength by 5 to 9% (p < 0.05 to p < 0.01), and muscular endurance by 55% (p < 0.01). Reductions in fat (p < 0.01) and gains in fat free mass (p < 0.05) averaged 3%. The females demonstrated greater improvements than the males. The majority of OCdts passed the representative military tasks, although females in some trades showed high failure rates. Forty six percent of OCdts sustained injuries in the first term, resulting in 5% of man days lost. No gender difference was found in injury rates. The least aerobically fit OCdts sustained more injuries than their fitter counterparts. In conclusion, there is scope for optimizing the effectiveness of the CC to enhance fitness and improve the focus of physical training on maximizing military task performance. PMID- 10377714 TI - Doctors five: African-American contract surgeons in the Spanish-American War. AB - The role played by African-American contract surgeons during the Spanish-American War has been a neglected aspect of the war's medical history. These men left their civilian practices, as did their white colleagues, to serve in the U.S. Army for relatively (even for the time) little compensation. After the war, as "civilian employees," they were not even eligible for a pension. Patriotism motivated their call to arms. PMID- 10377715 TI - A fugue-like state associated with diazepam use. AB - Diazepam is a long-acting benzodiazepine. Although diazepam is commonly associated with a variety of side effects, it is generally not believed to cause fugue-like states or retrograde amnesia. This report presents the case of an active duty patient who developed a brief fugue-like state with retrograde amnesia. This was associated with the short-term oral use of diazepam. There was no other apparent cause for his symptoms, which resolved within 24 hours after the diazepam was discontinued. This case suggests that short-term use of diazepam can lead to a brief fugue-like state with retrograde amnesia that has not been reported previously. PMID- 10377716 TI - Cerebral abscess after presumed superficial periorbital wound. AB - Penetrating wounds in the periorbital region may appear superficial and minor at first glance. The unique shape and thin bony roof of the orbit give these injuries a significant risk of associated intracranial penetration. This can initially be asymptomatic, and a high index of suspicion is essential to properly diagnose and treat these injuries. We report a case of an 8-year-old female who presented with delayed seizures from a frontal abscess resulting from such an injury. This article reviews the literature and discusses the appropriate management that should be used by emergency room and military physicians. PMID- 10377717 TI - Common peroneal nerve palsy in a UH-60 aviator. AB - A case of common peroneal nerve palsy in a UH-60 Blackhawk U.S. Army helicopter pilot is reported. A review of the literature revealed several reports of common peroneal nerve palsy, although there were no published reports of this injury secondary to performing flight duties in the UH-60 cockpit. A common practice among Blackhawk pilots is to brace the "collective" with their left knee, subjecting the common peroneal nerve to possible injury. This action should be considered as a possible cause of common peroneal nerve palsy in this select group of aviators. PMID- 10377718 TI - Multiple idiopathic left ventricular aneurysms in a Japanese woman. AB - Idiopathic aneurysms of the left ventricle (LV) are uncommon in Western society. Multiple idiopathic LV aneurysms are distinctly unusual and are rarely reported. As with aneurysms of atherosclerotic origin, these entities may be associated with chest discomfort, congestive heart failure, cardiac dysrhythmias, and thromboembolic phenomena. We present the case of a Japanese woman living in the United States with chest discomfort, ventricular arrhythmias, and a previous transient ischemic attack who demonstrated four discrete LV aneurysms on ventriculography. Extensive evaluation demonstrated no clear cause for these aneurysms. The patient was treated conservatively with medical therapy and has continued to do well without adverse clinical sequelae. PMID- 10377719 TI - [The relationship between Spanish and French neurology]. PMID- 10377720 TI - [Spanish study of quality of life in migraine (II). Profile of medication consumption and subjective efficacy]. AB - OBJECTIVES: The response to the different antimigraine medications is variable. In this study we have analysed the profile of prescription of these antimigraine medications, both preventive and symptomatic, by a group of spanish neurologists and examined the subjective efficacy of these compounds. PATIENTS AND METHODS: Neurologists from 7 hospitals in different spanish regions interviewed 305 patients (at least 40 per hospital) who met migraine diagnostic criteria. They used an ad hoc questionnaire in which the antimigraine medications, both symptomatic and preventive, taken by the patients, as well as their subjective response were registered. Patients with transformed migraine or tension-type headache more than 2 days per week were excluded. RESULTS: Analgesics, non steroidal anti-inflammatory drugs, ergotics and sumatriptan had been taken by 99, 69, 54 and 40% of the 305 interviewed patients, respectively. A subjective good response was refered to by 9% of patients who had taken analgesics, 23% of patients who had taken non-steroidal anti-inflammatory drugs, 39% of those who had taken ergotics and 63% of patients with sumatriptan. The current symptomatic treatment was: analgesics 34% of cases, non-steroidal anti-inflamatory drugs 26%, ergotics 13% and sumatriptan 63%. Regarding preventive treatments, 108 patients (35%) had been treated with calcium-antagonists, 87 (29%) with beta-blockers, 55 (18%) with amitriptyline and only 7 (2.2%) with valproic acid. The percentages of good responses to these drugs were: 55% for beta-blockers, 42% for calcium antagonists and 31% for amitriptyline. CONCLUSIONS: Our data confirm that analgesics are not efficacious in the majority of migraine patients and that the advent of sumatriptan has clearly improved the quality of migrane symptomatic treatment, even though about one-third of migraine patients do not respond to this drug. This study confirm that calcium-antagonists are the antimigraine preventive treatment most frequently prescribed in our country, even though their subjective efficacy is lower than that of beta-blockers. PMID- 10377721 TI - [Interobserver variation in the diagnosis of stroke]. AB - BACKGROUND: The variability in the diagnostic observations could be a problem both in the clinical and in the epidemiological field. Scarce data exist in the literature on this phenomenon, relatives to the field of the cerebrovascular illness, in their nosologic, semiologic and etiologic aspects. We pretend to study the interobserver variability in the diagnosis of the stroke, classifying it as absentee, present or possible in front of a given case. METHODS: 1. A questionnaire of 99 clinical cases has been elaborated, in order to be subjected to the evaluation of 10 neurologists (3 "junior", residents, 3 "senior" residents, and 4 "staff"); 2. The categories are: a) not stroke either transient ischaemic attack; b) probable stroke or transient ischaemic attack; c) transient ischaemic attack and d) stroke. 3. STATISTICAL ANALYSIS: by means of the statistic kappa of Fleiss (kappa) for several observers and several categories, determination of the standard error and 95% confidence intervals. RESULTS: In the diagnosis of the cerebrovascular illness: a) among all the observers: kappa = 0.49 (0.44-0.50); b) among staff: kappa = 0.51 (0.46-0.56); c) among residents: kappa = 0.47 (0.44-0.50). Global data in transient ischaemich attack diagnosis: kappa = 0.52 (0.49-0.55); in the diagnosis of the stroke: kappa = 0.57 (0.54 0.60); in the exclusion of stroke: kappa = 0.64 (0.62-0.67). CONCLUSIONS: With Fleiss' criteria, an acceptable to good agreement exists in the diagnosis of the stroke in base to the mere description of the clinical picture. This interobserver agreement is better if it is tried to exclude the stroke, and it could increase with the training of the neurologist. PMID- 10377722 TI - [Meningitis by Cryptococcus neoformans in patients with HIV infection]. AB - OBJECTIVE: To review a serie of patients with cryptococcal meningitis and immunodeficiency syndrome (AIDS) treated in our hospital in the last two years. PATIENTS AND METHODS: Retrospective study of 25 patients infected with the human immunodeficiency virus (HIV) and affected by Cryptococcus neoformans meningitis. The factors analysed were epidemiological data, clinical manifestations, biochemical and microbiological characteristics of cerebrospinal fluid (CSF), radiological abnormalities, treatment, adverse reactions and outcomes. RESULTS: Eighty-four percent of patients had less than 200 CD4/microliter. Cryptococcal infection was the AIDS defining illness in 24% of cases. Patients typically presented with neurologic symptoms such as: headache (88%), fever (68%) and somnolence (68%); 20% presented seizures and 28% focal deficits. There were no CSF biochemical alterations in 25% of them. CSF culture and indian ink stain were positive in 76%. CSF cryptococcal antigen test was positive in 68% of the cases. TC showed abnormalities in 48%. CSF of all patients treated with amphotericin B (AB) plus flucytosine (5FC) whose CSF culture was monitored became negative in the first two weeks, meanwhile those treated only with AB or fluconazol had negative control culture in 60% and 50% respectively. Six patients died within the initial 10 weeks. Death was due to bacterian sepsis in 3 patients and high intracranial pressure was the cause in 2 cases. One happened before treatment was administered. CONCLUSIONS: It's essential to consider the possibility of cryptococcal meningitis in patients infected with HIV and any compatible symptom regardless of CSF biochemical results and immunodepression level (CD4). Although our study was non randomized and so we can't propose a therapeutical schedule based on it, we can say that patients treated with AB plus 5FC showed an earlier conversion from positive to negative CSF cultures without more adverse reactions. PMID- 10377724 TI - [Cortico-basal degeneration: anatomical and functional neuroimaging]. PMID- 10377723 TI - [Pramipexol: a new dopaminergic agonist for the treatment of Parkinson disease]. AB - Pramipexol is a novel nonergot dopamine agonist which has high selectivity for intereacting with dopamine D2 receptors (especially with D3 receptor subtype). It has been effective in early Parkinson's disease as monotherapy and as adjunctive therapy with L-dopa in advanced stages of the disease. Clinical improvement can be observed after 3 or 4 weeks of treatment. The adverse events profile of pramipexol is similar, in general, to that of other dopamine receptor agonists, although it can be foreseen that pramipexol should not induce side effects related to the ergot chemical structure such as eritromelalgia, distal vasospasm, retroperitoneal fibrosis or pleural effusions. Nevertheless, the potential advantages of this promising dopamine agonist should be tested in well-designed prospective comparative studies with other available ergot and nonergot dopamine agonists. PMID- 10377725 TI - [Parkinson disease. Update in 1999]. PMID- 10377726 TI - [The problem of accurate diagnosis of Parkinson disease]. AB - A definite diagnosis of Parkinson's disease cannot be made solely on clinical grounds. None of the cardinal signs (tremor, rigidity, or bradykinesia) is entirely specific, including asymmetric presentation. A favourable response to levodopa may be found in different degenerative parkinsonisms or there may be drug-induced dyskinesias and fluctuations. "Probable" or "possible" as diagnostic categories depend on the presence or one or more cardinal signs, absence of potential etiologic factors (exposure to antidopaminergic drugs, among others), and atypical clinical manifestations early in disease course as dementia, falls and instability, erectile and micturition disturbances and unilateral neglect. Preclinical detection (positron emission tomography and molecular genetics in some familial forms) is a desirable goal with a view to neuroprotection. We emphasise some prodromal and early complaints as well as clinical skills to detect cardinal signs during early. PMID- 10377728 TI - [Current therapeutic problems. Solutions]. PMID- 10377727 TI - [Pharmacologic treatment in Parkinson disease]. PMID- 10377729 TI - [New pharmacologic strategies for the treatment of Parkinson disease]. AB - The dopamine precursor, levodopa, and the synthetic dopamine agonists are drugs widely used to alleviate the motor symptoms of idiopathic Parkinson's disease. Recently, several molecules which act on different pharmacological receptors implied in Parkinson's disease pathophysiology, have been developed to be applied as a treatment for its motor and cognitive symptoms. This paper discusses these novel therapeutical strategies, their mechanism of action, and their potential applications. To facilitate its lecture, the article has been structured in several sections. In the first section, drugs acting on dopamine metabolism are described, in the next part recent studies on new dopamine agonists and drugs acting on non-dopamine neuronal receptors are analysed, and finally the role of trophic factors for the treatment of Parkinson's disease is discussed. PMID- 10377730 TI - [Pathophysiological bases, clinical results and indications for surgical treatment in Parkinson disease]. AB - We review the present status of surgery for Parkinson's disease. Surgical options for Parkinson's disease are rapidly spanding. The main objectives of surgical techniques are to restore the dopaminergic deficit in the striatum (transplantation) and to normalize the neuronal activity of the subthalamic pallidal circuit (pallidotomy and deep brain stimulation). Whereas cell transplantation is still considered an experimental procedure, ablative procedures and deep brain stimulation are widely used. Both types of surgical procedures are supported by strong scientific data. However, much work remains to be done in order to understand several aspects not clearly elucidated at present. The results and current indications for pallidotomy and deep brain stimulation are analyzed. PMID- 10377731 TI - [Parkinson disease and cognition]. AB - Parkinson's Disease patients may have cognitive deficits, which may vary from mild focal specific deficits to global dementia. Executive functions, working memory, visuoespatial functions and internal control of attention are the main affected cognitive areas. The dopaminergic hypothesis suggests that dopaminergic transmission is involved in most altered cognitive processes. However, other neuronal systems are probably implicated, and the improvement of the cognitive function after dopaminergic replacement is incomplete and not seen in all cognitive tasks. The prevalence of dementia in Parkinson's disease is estimated in 15-25%. The pathologic hallmarks may be similar to that seen in Alzheimer's disease. However, in Parkinson's disease patients pure subcortical alterations are able to produce a severe cognitive impairment, such as lost of dopaminergic neurones from substantia nigra to caudate nucleus and frontal areas. Comprehension of the relationship between depression and dementia in Parkinson's disease patients is incomplete. Parkinson's disease patients with depression show poor neuropsychological performance, especially in frontal tasks, but it is unclear whether depression can be considered a risk factor for the development of dementia in Parkinson's disease. PMID- 10377732 TI - [Cardiac electrophysiological effect and clinical use of adenosine]. AB - A review is provided of the present state of knowledge relating to the cardiac electrophysiological effects of adenosine at ion channel and clinical levels. It is emphasized that intravenous adenosine is highly effective in terminating sinus node, atrioventricular (AV) nodal and AV-reciprocating reentrant tachycardias. In the course of an account of the antiarrhythmic indications of adenosine therapy, the main aspects of the diagnostic use of the nucleoside are briefly discussed. It is pointed out that adenosine treatment may possibly be accompanied by the occurrence of atrial and ventricular proarrhythmias, for at pharmacological dose of 6-12 mg this endogenous cardioprotective and antiarrhythmic agent is not devoid of side-effects. PMID- 10377733 TI - [Interferon therapy of chronic viral hepatitis in Hungary: 5-year experience. A multicenter study]. AB - In Hungary over the past 5 years more than thousand patients with chronic viral hepatitis have been examined and included in a treatment program with interferon (IFN) at 16 major hepatology centers, using unified diagnostic and therapeutical criteria. Authors give an account of their experiences on the clinical features of patients with chronic viral hepatitis and report the results of the treatment with IFN. According to the rules and availability of IFN for patients with chronic viral hepatitis in the country, virtually the entire Hungarian population with this diseases who required IFN therapy have been included. A total of 94 patients suffered from hepatitis B virus (HBV) infection, in addition 11 HBV + hepatitis Delta virus (HDV), 24 HBV + Hepatitis C virus (HCV) related liver disease, and 993 had chronic hepatitis C. IFN therapy for chronic HBV hepatitis consisted of IFN 5 MU thrice weekly for 6 months, and resulted in 33% seroconversion and sustained remission with 14% HBsAg clearance. For chronic hepatitis C treatment protocols (dose of IFN and duration of therapy) have changed with the time (from a weekly dose of 3 x 3 MU IFN for 6 months, to 3 x 3 MU for 12 months), and even a combination with ribavirin has been introduced. Although the therapeutic results showed a gradual improvement form a 13% sustained response over 22% in the first and second periods, respectively, differences were most significant with the advent of the combination therapy, that resulted in 36% remission rate. Only fibrosis in histology and baseline pretreatment HCV-RNA level appeared as predictors of response in chronic hepatitis C. Neither age nor gender did influence the outcome, but longer duration of treatment and higher total dose of IFN resulted moderately higher sustained remission rates. The experiences are in accordance with findings of suboptimal efficacy of IFN monotherapy reported worldwide and emphasize the need of seeking for newer and combination therapeutic modalities for these chronic viral diseases. PMID- 10377734 TI - [Experience with combined interferon alpha-2b and ribavirin in the therapy of chronic hepatitis C. Experience with one-year therapy of 100 patients. Multicenter study]. AB - It has been established that the long term interferon therapy in patients with chronic hepatitis C is able to produce sustained remission only in about 20 per cent of the cases. According to the newest data the combined interferon and ribavirin therapy significantly increases the remission of patients in naive, non responder or relapsed cases. Clinical remission was confirmed by enzyme activity of alaninamino transferase (ALT) and HCV-RNA-PCR tests. In order to get exact data of the remission rate and the symptom free period, a prospective multicenter study has been introduced in Hungary. Ten leading hepatologic units have been involved into the trial. Till now the combined therapy with interferon-alfa-2b (3 MU, three times a week) and ribavirin--(1000-1200 mg daily) for one year has been finished in 100 cases with chronic hepatitis C. The mean value of ALT activity decreased near to the normal level, in 58 patients it was in the normal range. Side effects with mild or moderate grades have been found in 31 cases. The interim report of this multicenter study confirm the efficacy of this combined therapy in chronic hepatitis C. PMID- 10377735 TI - [Transbronchial biopsy in diffuse infiltrative lung diseases]. AB - 224 transbronchial biopsies were made between 1990 and 1997 in 208 patients suffering diffuse, bilateral, disseminated pulmonary diseases with unidentified origin. The examinations were carried out by flexible bronchoscope with X-ray control after local anaesthesia. The obtained patterns yielded mucous membrane in 15, intact alveoli in 19 percent. In 25 percent of biopsies (57 cases) definitive diagnosis were verifiable. The histological examinations verified the diagnosis of sarcoidosis, hemosiderosis, TB, malignancies, eosinophil pneumonitis etc. In 71 biopsies (62 cases) according to histological opinion were pulmonary fibrosis or some synonyms of these (alveolitis, interstitial fibrosis). From these latter cases the documentation of 18 patients were insufficient, but the data of 44 cases were available. 7/44 carcinosis, 3/44 inactive TB, 8/44 regressive X-ray patterns were verifiable in this group. The idiopathic pulmonary fibrosis as final diagnosis only in 18/44 (41%) cases were established. Further diagnostic procedures are needed if the clinical data, the results of HRCT and histological examinations are not in correlation. PMID- 10377736 TI - [Principles of drug therapy of malignant tumors: the role of malignant progression in the choice of effective drugs]. AB - The progress that has been achieved in the development of antitumour drugs and the management of the untoward side effects both contributed to the increasing importance of drug-therapy beside surgery and radiotherapy in cancer treatment. The treatment of the micrometastasis which appears very frequently already at the time of the diagnosis and also the control of the metastatic progression represent the main importance of drug therapy in cancer patients. In spite of the numerous clinical trials indicating the usefulness of drug therapy both as adjuvant in the management of the primary tumours and in the treatment of metastatic tumours there are certain reservation against chemotherapy in medical circle. It is noteworthy that at the present time the strategy of cancer-therapy is subject of considerate changes. Beside to achieve cure by eradication of the tumour cells it has been recommended in various oncological center that stabilization of the malignant disease and to offer a good quality of life for the patients should also be the aim of the therapy. The purpose of this communication is to present those factors which are necessary to consider in the planning of anticancer drug therapy. Certainly to achieve cure or to improve the quality of life of cancer patients it is important to select the most appropriate drug (cytostatics, hormones, biological response modifiers or agents improving the quality of life) and treatment schedule. Since antitumour drug therapy must be classified as active, palliative/active, palliative and supportive/terminal treatments the present survey gives emphases on the underlying role of malignant progression before deciding the type of treatment. PMID- 10377737 TI - [Rapid hemoglobin A1c determination (a new possibility in diabetes care)]. AB - To assess the long-term metabolic control, immunochemical method was used for hemoglobin A1c (HbA1c) determinations in diabetic patients. The use of DCA 2000 device (Bayer) resulted in immediate (< 6 min) HbA1c values. The reproducibility of this method was acceptable (within-run coefficients of variations were 3.48% and 4.80%). A close, linear correlation (r = 0.974; p < 0.001; n = 106) between HbA1c-values measured simultaneously by DCA 2000 and DIAMAT (Bio-Rad, method: high pressure liquid chromatography) was observed in diabetic patients. The new immunochemical method proved to be simple and reliable. The immediate (within 6 min) result makes the therapeutic decision easier during the care of diabetic patients. PMID- 10377738 TI - [The community hospital and the poorhouse of Buda in the 16th century]. PMID- 10377739 TI - [Clinico-pathology of lobular breast cancer]. AB - 997 operations were performed because of malignant breast lesions at the National Institute of Oncology during a two-years period (1990-1991). Histologically 94 tumours proved to be invasive lobular cancer. Comparable data were available for 89 patients; a questionnaire was used for data collection. Analysing these cases, the authors discuss the clinicopathology of lobular cancer and current practice in its treatment. The mean age of the patients was 57.1 years, which in accordance with the literature. No synchronous contralateral tumour was observed in the studied patient group. 4.9% of the cases proved to be multifocal, what is considerably less than that reported in the literature. It is a frequent matter of debate in many papers whether mastectomy should be indicated if the multicentricity of the tumour is only suspected. The features of the studied group may also emphasise this question, and further investigation is needed for moderating the surgical radicality. The rate of large tumours and the mean tumour diameter was lower than suggested by the literature. The rate of positive axillary lymph nodes was higher than in reports of other centres. The authors underline the necessity of axillary block-dissection even in case of operations with decreased radicality. Beside lymph node metastases, invasive lobular cancer produces haematogenous metastases with a higher probabilty than other breast cancers. This observation is supported by our study, too. PMID- 10377740 TI - [Clinical experience with surgical management of atrial myxoma]. AB - Clinical data (symptoms, diagnostic tools, surgical and histological findings, postoperative course and present status) of 26 patients with cardiac myxomas were reviewed. All myxomas were of left atrial localization. The diagnosis was confirmed by cardiac catheterization and angiography (until late seventies) and echocardiography. The delay between diagnosis and surgical management was minimalized. There was one early postoperative death. During follow-up period, one patient died, most patients live without symptoms. No recurrences have been found by echocardiographic follow-up examinations. Late consequences can be revealed by careful follow-up and regular echocardiographic studies. PMID- 10377742 TI - [Regulation of data protection in health care]. AB - Computers are widely used in different fields of health care, the traditional administrative systems being replaced by softwares. However, the use of computer technology in health care has not yet brought to a definite improvement in the field of data protection and data security, what is more, new problems have been revealed. The legal regulation of data protection in Hungary dates back to 1992. There are technological norms, different standards, guidelines and the regulation of data protection in health care on a sectoral level. What is missing is the security regulations of information systems in the institutions. Joining the European Community requires the implementation of an adequate data protection being able to ensure the security of personality rights also in the field of health care. The use of an up-to-date insurance card would strengthen the data protection and security functions offering the possibility for introducing an almost "paperless" administrative system in health care. PMID- 10377741 TI - [Incidence of type 1 childhood diabetes in Hungary (1978-1997). Hungarian Committee on the Epidemiology of Childhood Diabetes]. AB - The incidence of Type 1 diabetes in 2417 children aged 0-14 years was studied between 1978 and 1997. Statistical analysis of the data in the twenty-year period showed a steady increase in incidence of Type 1 diabetes with an average rate of 4.8% (95% CI 4.0-5.5) per annum. The incidence rate was lowest in children aged 0 to 4 years and the highest in the age group 10 to 14 years. This increase in incidence was similar for all age groups, but it was most pronounced in the 10 to 14 year-old age group. One hundred and twenty one of the 1239 families (9.7%), one of parents was also reported to have Type 1 diabetes and in 3 (0.2%) families both parents had Type 1 diabetes. Twenty two (2.3%) of the diabetic families had a sibling with Type 1 diabetes. The nearly three-fold increase in incidence during the last two decades (3.8 per 100,000 per year in 1978; 10.7 per 100,000 per year) was probably due to environmental factors, while the high degree of familial clustering emphasises the role of genetic factors in the etiology of Type 1 diabetes. PMID- 10377743 TI - [Balneotherapeutic institutes based on the "Graefenbereg model" in Hungary 1848 1849. (Vinzenz Priessnitz (1799-1851), pioneer of empiric medicine, born 200 years ago)]. PMID- 10377744 TI - [A wheat germ preparation and its possible action]. PMID- 10377746 TI - [Comment of the use of positron emission tomography]. PMID- 10377745 TI - [Let us help retired persons by reducing medical costs]. PMID- 10377747 TI - Multiple genes governing biological functions in the genetic backgrounds of laboratory mice and Asian wild mice. PMID- 10377748 TI - Overview of transgenic and gene knockout mice. PMID- 10377749 TI - Alkylation carcinogenesis in mice with altered levels of DNA repair methyltransferase. PMID- 10377750 TI - Cancer predisposition in mutant mice defective in the XPC DNA repair gene. PMID- 10377751 TI - Myeloid leukemia: disease genes and mouse models. PMID- 10377752 TI - Genetic and epigenetic susceptibility to endogenous retrovirus-induced lymphomas in SL mice. PMID- 10377753 TI - Inheritance of pulmonary adenoma susceptibility in mice. PMID- 10377754 TI - TSC2 gene mutant (Eker) rat model of a Mendelian dominantly inherited cancer. PMID- 10377755 TI - Apc gene knockout mice as a model for familial adenomatous polyposis. PMID- 10377756 TI - A rat model system for predisposition to stomach cancer. PMID- 10377757 TI - Genetic and hormonal regulation of murine hepatocarcinogenesis. PMID- 10377758 TI - Genetics of skin tumor promotion. AB - Cancer development is a multistep process that involves both the activation of protooncogenes and the inactivation of tumor suppressor genes. Furthermore, both epidemiological and experimental data indicate that a third class of genes, tumor susceptibility genes, control the propensity to develop carcinogen-induced tumors. Recent studies suggest that tumor susceptibility is determined by the combined effect of both sensitivity and resistance genes. The mouse skin model of multistage carcinogenesis is an excellent paradigm in which to study the genetics of cancer susceptibility. This is particularly true with regard to tumor promotion, a process that occurs in other organs and species including humans. We have studied the genetics of tumor promotion susceptibility in the mouse two stage skin tumor model using crosses between sensitive DBA/2 or C3H and resistant C57BL/6 mice. Our results suggest that TPA promotion susceptibility is a multigenic trait. We have tentatively mapped one tumor susceptibility locus, Psl1, to mouse chromosome 9 and are currently identifying and characterizing candidate tumor susceptibility genes that map to this chromosomal region. The multistage model of carcinogenesis in mouse skin has, for more than 50 years, provided a conceptual framework from which to study the carcinogenesis process. Many concepts now currently applied to other tissues and model systems were originally derived from the mouse skin model. Because tumor promotion is an important component of carcinogenesis in humans, the identification of genes that modify response to tumor-promoting stimuli would be a significant advancement in our understanding of the genetic basis of susceptibility to cancer. PMID- 10377759 TI - The genetic components of susceptibility to breast cancer in the rat. AB - The rat is an extremely valuable model for studies of inherited susceptibility to breast cancer because the characteristics of rat mammary cancer and human breast cancer are so similar. There are now several rat models for studying sensitivity versus resistance, or cell autonomy versus non-cell-autonomy, for spontaneous and induced mammary cancers. It is known that the tumor-resistant Cop [20, 21] and WKy [8] strains carry dominant resistance genes that inhibit both spontaneous and induced mammary tumors. The WF and SD strains are known to carry dominant sensitivity genes that appear to increase susceptibility to induced but not spontaneous mammary tumors. The presence of both resistance and sensitivity genes in the Cop strain is intriguing, and provides a unique model for studying the interactions of both types of genes. It appears that the resistance genes together are at least partially dominant over the sensitivity gene in this model since the F1 rats develop only a few tumors. Yet another strain, the F344, has an intermediate sensitivity and has been shown to carry neither sensitivity or resistance genes. Thus, all these models and data indicate that sensitivity genes are not necessary for the development of mammary tumors, and neither are they sufficient. However, loss of resistance gene function is necessary but is not sufficient for mammary tumor development. Studies have shown that the sensitivity and resistance genes act directly within the mammary epithelial cells rather than globally in the rat. The products of these genes also do not appear to act at early steps in the carcinogenic process because there have been no observed effects of these genes on carcinogen metabolism or DNA adduct formation. It would appear that these genes act at later stages of mammary carcinogenesis. Identification and isolation of these genes should aid our understanding of the inherited components of human breast cancer. With the increasing availability of genetic markers and large-insert libraries for the rat genome, genetic and physical mapping studies are now a reality for the genes involved in mammary carcinogenesis of the rat. Such studies have already revealed the multigenic nature of this cancer, supporting the idea that the limited penetrance of BRCA1 and BRCA2 in human breast cancer is due to loci that modify the effects of the sensitivity genes. Assuming that human homologues of the Mcs genes exist, cloning the genes and defining the human homologues may provide a way to identify the risk for breast cancer development in women. Analysis of the function of such genes may also lead to the development of new drugs for chemoprevention and/or therapy of this lethal disease. PMID- 10377760 TI - [Laparoscopically-assisted endoscopic polypectomy]. AB - OBJECTIVE: Make it possible to implement safe polypectomy of polyps of the large intestine with a diameter at the base greater than 1 cm located in the intestinal lumen above the peritoneal fold. METHOD: After laparoscopy under general anaesthesia endoscopic polypectomy is performed under laparoscopic control. The large intestine is sutured to the site of the base of the removed polyp using laparoscopic technique. RESULTS: Removal of large polyps of the large bowel with a base diameter greater than 1 cm without the need of laparotomy. CONCLUSION: Laparoscopically assisted polypectomy is a safe and effective method for the removal of large bowel polyps with a large base. PMID- 10377761 TI - [Peritonism]. AB - The authors discuss clinic symptoms, reminding of acute abdomen, which called peritonism. Two patients admitted and treated at the 1st Department of Surgery are presented. These patients meet the criteria of the category of peritonism. PMID- 10377762 TI - [Videothoracoscopy and video-assisted thoracic procedures in the diagnosis and therapy of diseases of the thoracic cavity (experience with 155 procedures)]. AB - Videothoracoscopic procedures and video-assisted thoracic surgery (V.A.T.S.) are up-to-date procedures in thoracic surgery. The authors evaluated results from 155 cases. They reviewed especially indications, complications and results. The most frequent indications were spontaneous pneumothorax, fluidothorax, interstitial lung disease and lung carcinoma. Staging was main procedure in cases of lung carcinoma. Complications were observed in 14 cases (9%). Of these complications 5 developed at the time of surgery (3.2%) and 9 were postoperative (5.8%). PMID- 10377763 TI - [Dynamics of the sliding screw]. AB - Fractures of the upper end of the femur are treated by the authors since August 1993 by the method of a dynamic gliding screws. This method is considered sparing and economical. The operation is implemented as urgent with a screen of antibiotics and anticoagulants in order to reduce postoperative complications. As a matter of routine intensive early rehabilitation is ensured and early return of the patient to his normal environment. Between August 1, 1993 and August 31, 1998 at the authors clinic 259 osteosyntheses were made using a dynamic gliding screw. There were 10 reoperations. In three instances the reason was incorrect surgical technique, in seven instances unsuitable indication of osteosynthesis in an arthrotic joint with impaired nurture in a severely dislocated fracture of the subcapital type. Reoperation was implemented twice by the original technique, eight times by replacement of the hip joint [4, 6]. For successful treatment of patients with fractures of the upper end of the femur it is essential that the department should have an X-ray amplifier, instruments for a dynamic gliding screw, a gamma and reconstruction nail and a hip joint replacement. PMID- 10377764 TI - [Surgical treatment of benign tumors of the thyroid gland]. AB - Based on 22-year experience with surgery of 2107 benign thyroid tumours and data in the literature the authors evaluate some controversial problems from this sphere of thyrology pertaining to the definition and classification of benign tumours, their diagnosis, treatment and subsequent follow-up. Modern diagnostic procedures indicate that the actual number of true benign tumours is smaller than reported in some statistics and the assumption of monoclonal development of tumours is one of the most fundamental characteristics. The authors consider total lobectomy as minimal surgery, in case of uncertainty as regards possible malignancy further procedures depend on final histological examination. If the final finding alters the diagnosis from an originally benign tumour to a malignant one, the authors recommend a two-stage procedure to implement total thyroidectomy. In the authors' opinion total lobectomy is sufficient also in some variants of follicular adenoma--Hurtle's adenoma or atypical adenoma. Then however subsequent dispensarization is essential. After a partial operation (lobectomy) it is important to ensure substitution with thyroid hormones to suppress the function of TSH. PMID- 10377765 TI - [Malignant lymphomas of the thyroid gland]. AB - The authors evaluate their experience with malignant lymphomas of the thyroid gland, recorded in 22 patients from a total of 4501 operated on account of thyroid diseases in the course of 22 years. Although thyroid lymphomas are rare, their number is increasing. In localized findings curative procedures can be used, possibly supplemented by operations of the lymph nodes. The method of choice is still radiotherapy and surgery, in particular in lesions restricted to the thyroid gland. The role of chemotherapy is still controversial. PMID- 10377766 TI - [Quo vadis the azygo-portal disconnection]. AB - Based on experience at their department since 1983 the authors analyze the position of the azygoportal disconnection in the treatment of complications of portal hypertension. During the observation period the number of operations declined due to extensive use of endoscopic diagnosis and treatment. However the azygoportal disconnection still remains part of surgical intervention in indicated cases, in particular in secondary hypersplenism and profuse haemorrhage. PMID- 10377767 TI - [Cystic degeneration of the popliteal artery adventitia]. AB - The author presents the case-history of a typical clinical picture of a rare vascular affection--cystic degeneration of the adventitia of the popliteal artery in a young man. The diagnostic conclusion was reached by means of duplex sonography and angiography of the extremity, the condition was treated by reconstruction surgery. With regard to its long-term effect the author prefers it to some non-surgical procedures which include in particular transluminal angioplasty or sonographically assisted needle aspiration of the cyst. PMID- 10377768 TI - [A preventive ostomy in low rectal resections]. AB - Dehiscence of the anastomosis after low resection of the rectum is a serious complication with the possible development of sepsis and multiorgan failure. This complication is reported in 10-20% of operations. The incidence of dehiscences can be reduced when known principles of preoperative care are respected but in particular by correct surgical technique. The serious character of this surgical complication can be reduced by a primary derivative stomy. Based on their experience the authors recommend to implement a preventive ileostomy in low resections of the rectum which can be closed already two weeks after the primary operation. PMID- 10377769 TI - [Central rupture of the liver in multiple injuries]. AB - Authors evaluate clinical material of polytraumatized patients operated on account of liver injury during the last 15 years. The contribution of new imaging techniques is analyzed, especially of spiral computed tomography, for the diagnosis of liver injuries and associated injuries of the abdomen and retroperitoneum. These techniques enable in stabilized patients with the trauma I, II injuries and in some cases of grade III injuries to prefer conservative treatment. During the last 3 years the authors treated 5 polytraumatized patients with central liver rupture. Consistent with experience of major trauma centres in USA and Switzerland, criteria for conservative treatment are outlined, especially in patients of younger age groups. PMID- 10377770 TI - [Personal experience with surgical treatment of rupture of the Achilles tendon]. AB - The authors summarize 13 years experience with surgery of ruptures of the Achilles tendon. They describe the therapeutic and rehabilitation procedures and results recorded in 50 operated patients. By restricting rigid fixation, treatment can be substantially shortened. PMID- 10377771 TI - [The compartment syndrome in injuries of the upper extremity in children]. AB - In the Centre for Child Traumatology in Brno the authors followed up during the last 10 years 22 patients with suspected compartment syndrome of the upper extremity. Incisions of intrafascial spaces were made in 12 patients-9x on the forearm and 3x on the hand. Another 10 patients had conservative treatment. The therapeutic results are with the exception of one female patient with developed Volkmann's contracture very satisfactory, the sequelae are only cosmetic. Compartment syndrome is a serious acute condition in child traumatology. Permanent sequelae of this syndrome can be prevented above all by early diagnosis and adequate therapy. It is therefore important to consider the possible development of this serious complication. PMID- 10377772 TI - [Ethical problems in organ transplantation]. AB - Organ transplantation is an accepted therapeutic method with good results, but it is connected with many not only medical but also ethical problems. One of the most important problems is the donor programme. In cadaverous donors the main ethical and legal question is the decision who can issue the consent with organ retrieval; in living donors it is the problem of motivation and financial compensation. Allocation of organs with low compatibility or from non-ideal donors, and the recipient's consent in these cases may involve difficult decisions. PMID- 10377773 TI - [Inflammatory (infectious) complications in arteriovenous fistulae for hemodialysis]. AB - Angioaccess procedures for hemodialysis over a 12-year period were retrospectively reviewed to ascertain the frequency of infectious complications. A total of 571 angioaccess procedures were performed including 88 e polytetrafluoroethylene (ePTFE) grafts. Early infectious complications occurred only in 5 autologne fistulae. Late infectious complications developed in 2.3% autogenous fistulae and in 7.95% ePTFE grafts. Infected arteriovenous fistulae can be the cause of further complications (thrombosis, bleeding, aneurysm, sepsis) and call for surgical repair. PMID- 10377774 TI - [The compartment syndrome in injuries of the lower extremity in children]. AB - A total of 22 patients with suspected compartment syndrome in the lower extremity were treated over the period of the last ten years at the Paediatric Traumatological Centre in Brno. CS was confirmed in 15 cases of these patients and incisions of intrafascial areas were performed in the course of treatment, 13 times on the middle leg and twice on the thigh. A conservative procedure was selected in 6 cases when diagnosis was not clearly confirmed: twice on the foot and 4 times on the middle leg. In diagnostics and treatment we definitely prefer permanent monitoring of intrafascial pressures and performance of adequate fasciotomies. Therapeutic results revealed to be good in implementing a correct procedure. Prevention of the iatrogenic ratio of this serious syndrome caused particularly by late assessment of diagnosis is important. PMID- 10377775 TI - [Tragic results of an untreated compartment syndrome of the lower extremity]. AB - Despite its notorious extensive consequences compartment syndrome is generally underrated high risk states are not adequately monitored and treated and therefore there is a number of seriously injured child patients. Late surgical intervention and incorrect therapy are responsible for consequences and incomplete recovery. The authors present the case of a 9-year-old boy treated for a diaphyseal fracture of the thigh by vertical traction, where compartment syndrome developed during treatment and did not receive adequate attention. A combination of the incorrect procedure and the insufficient monitoring of the patient led to his permanent disability. PMID- 10377776 TI - [Preventing the development of secondary infections in severe acute pancreatitis]. AB - Severe acute pancreatitis is in its first stage characterized by the syndrome of systemic inflammatory response (SIRS). The extent of activation of the mediator defense cascade of the organism depends on a hitherto unknown primary insult which cannot be influenced by therapeutic care. The cause of the development of sepsis or multiorgan failure is the supraliminal action of secondary insults (ischaemic reperfusion syndrome, hypoxia, impaired physiological function of the gut). By early and aggressive therapeutic action the effect of these adverse factors can be markedly reduced. Then also better therapeutic results in severe acute pancreatitis may be expected. PMID- 10377777 TI - [Aberrant mammae, personal experience]. AB - The authors present information of a group of 63 patients (60 women and 3 men) treated in 1976-1998 where supernumerous aberrant or accessory mammary gland tissue was found. The diagnosis in the majority of the patients was based on histological examination. The tissue of an aberrant mammary gland is subject to the same physiological influences as the mamma proper and can be affected by the same pathological processes--benign and malignant. This is the reason why the authors recommend early extirpation. PMID- 10377778 TI - [Differential diagnosis of pain in the right lower abdominal quadrant--case reports]. AB - In the submitted case-records the authors present interesting peroperative findings--a coprolith obturing the lumen of the appendix, the torsion of an ovarian cyst and torsion of the omentum during diagnosis of acute appendicitis. The authors discuss the reliability of preoperative sonography for the diagnosis of pathological conditions in the appendical region which despite its falsely negative results are considered by the authors a useful method in the differential diagnosis of pain in the right iliac fossa. PMID- 10377779 TI - [New entities and pitfalls in the diagnosis of soft tissue tumors]. AB - In the presented review the authors described clinicopathological entities which were defined in soft tissue tumour pathology in the last ten years. They briefly discuss some new aspects of already well known soft tissue tumours and present their own diagnostic and published experiences with these lesions. PMID- 10377780 TI - [Surgery of a large ventral hernia using Gore-tex]. AB - Severe intraabdominal sepsis, i.e. perforating peritonitis or necrotizing pancreatitis with complications sometimes call for very aggressive surgery such as the method of temporary closure of the abdominal cavity with repeated revisions. This frequently leads to the development of extensive ventral hernia. The latter develops as a rule at a site where it is impossible to suture the aponeurotic part of the abdominal wall (it is too retracted or lacking) and the use of a synthetic mesh for its reconstruction is not possible for some reason. After elimination of the temporary closure thus only resuture of the skin and subcutaneous layer is made. The author demonstrates on two cases one of the possible operations of a thus developed major hernia, where a Gore-tex patch was used. This material meets best the demands laid on a permanent "substitute" of the abdominal wall. Its wider use is unfortunately prevented by its high price. PMID- 10377781 TI - [Temporary closure of the abdominal cavity--surgical technique]. AB - Temporary closure of the abdominal cavity can be achieved by different means. The disadvantage of original devices (Ethzip, Velcro) is as a rule their high price. The author presents his own technique of temporary closure of the abdominal cavity which meets all contemporary demands laid on this method, while it uses materials of Czech origin the price of which is incomparably lower than that of foreign devices. He uses as a basis worldwide findings as well as his own long standing experience with this method in the treatment of severe forms of haemorrhagic-necrotizing pancreatitis and diffuse peritonitis of varying etiology. The method was used at the Department of Surgery of the Faculty Hospital in Plzen since 1990 with favourable results in more than one hundred patients. PMID- 10377782 TI - [A technique for treatment of extensive defects of the abdominal wall with gastrointestinal fistulae]. AB - Under certain extreme conditions in abdominal surgery, such as in septic complications of resections of the GIT with dehiscence of the anastomosis the only possible surgical approach is classical laparostomy. In the latter, contrary to the technique of temporary closure, the abdominal cavity is left open. After control of the sepsis usually the greatest problem is treatment of the defect in the abdominal wall with a fistula of the GIT. Unfortunately special devices for the treatment of such wounds cannot be used in all cases. At the Department of Surgery in Plzen such situations are resolved by suction drainage led beyond the defect in the abdominal wall which is then covered with an incision foil. In this way treatment of the patient is greatly simplified. PMID- 10377783 TI - [Neuronal cell death--what we can see and what we cannot]. AB - Recently several responsible genes for hereditary neurodegenerative disorders were identified. In some of them the gene products were found to be aggregated. In the case of Alzheimer disease beta protein and apolipoprotein E accumulated in senile plaques. In CAG repeat diseases the polyglutamine aggregates in neuronal nuclei. More recently alpha synuclein accumulates in Lewy bodies in Parkinson disease and tau protein accumulates in NFT of hereditary frontotemporal dementia with tau mutation. Those results suggested that the responsible gene products accumulates in the lesion which the products involve in. However, presenilin which is one of the genes for familial Alzheimer disease accumulates in NFT and on the other hand its mutation changes the production ratio of beta 1-42/40, suggesting that the abnormal gene products not simply accumulate the lesion that it involved. The gene products accumulate in different lesions such as in nuclei of polyglutamine diseases, extracellular plaque and cytoplasm of prion disease and extracellular plaques in Alzheimer disease. Some of them are ubiquitinated and some of them are not. Thus the accumulating process in these disorders seems apparently same but is essentially different. We should study more precisely each pathological process of those disorders. PMID- 10377784 TI - [Neuronal cell death in neurodegenerative disorders and oxidative stress]. AB - Mechanisms of the process of neuronal degeneration in neurodegenerative disorders such as amyotrophic lateral sclerosis (ALS), Parkinson's disease (PD), and Alzheimer's disease (AD) remain unsolved. Oxidative stress might be a possible mechanism of neuronal cell death. Glutamate is an excitatory amino acid and its excessive release can cause intracellular calcium influx, activation of calcium dependent enzymes such as nitric oxide (NO) synthase (NOS), and production of toxic oxygen radicals. Excessive release of glutamate, therefore, can be used as a model of experimental oxidative stress. Continuous exposure to low levels of glutamate potentiates selective motor neuronal death mediated by NO, which inversely protects nonmotor neurons through the guanylyl cyclase-cGMP cascade. Mesencephalic dopaminergic neurons are resistant to cytotoxicity induced by NO. The protecting mechanism from NO neurotoxicity in dopaminergic neurons is based on inhibition of conversion of NO to peroxynitrite anion, and is possibly due to suppression of superoxide anion production. Dopamine D 2 agonists provide protection mediated not only by the inhibition of dopamine turnover but also via D 2-type dopamine receptor stimulation and the subsequent synthesis of proteins that scavenge free radicals. In addition, nicotinic receptor stimulation may be able to protect neurons from oxidative stress induced by A beta. PMID- 10377785 TI - [Transporter and neurological disease]. AB - There are two types of neurotransmitter transporters whose structure is known. One is a glutamate transporter family. The glutamate transporter takes up glutamate to terminate neurotransmission and to prevent neuronal cell death. The transporter may play important roles of cause and development in the neurological diseases. Another type is a catecholamine transporter family with 12 transmembrane domains. Dopamine transporter of the family takes up 1-methyl-4 phenylpyridinium (MPP+) and causes dopaminergic cell death. PMID- 10377786 TI - [A Parkin gene (PARK 2) and Parkinson's disease]. AB - We identified a PARK 2 (AR-JP) family with a patient presenting with homozygous deletion of D 6 S305--a marker within the 17cM region for PARK 2 locus. Markers surrounding D 6 S305 which are mapped 0 cM apart from D 6 S305, were not deleted, indicating that PARK 2 gene is located extremely close to D 6 S305. Exon search in the inserts with average size of 100 kb of BAC clones, which harbor D 6 S305, led us to find the exonic sequences which was subsequently proved to be exon 7 of the Parkin gene. From this exon sequences, full-length cDNA was isolated, and BAC contig covering Parkin gene was generated. Homozygous deletions or frame-shift mutations in the Parkin gene were found in the patients with AR-JP/PARK 2, revealing that a loss-of-function of Parkin gene is responsible for AR-JP/PARK 2. Our findings indicate that constant production of Parkin protein is essentially required for maintaining the survival of nigral neurons. One attractive hypothesis is that Parkinson's disease and AR-JP/PARK 2 might share a common effector pathway for nigral neuronal death. In this scenario, as PARK 2 is not accompanied with Lewy body formation. Parkin might act at or downstream of synuclein aggregation, which has been recently implicated as a trigger event for neuronal death in Parkinson's disease. In any case, identification of functional targets of Parkin protein will give us an important clue to identify downstream events of neuronal death which is activated by inclusion body formation. PMID- 10377787 TI - [Neuronal kinases in glial cytoplasmic inclusions in patients with multiple system atrophy]. AB - Glial cytoplasmic inclusions (GCI) occur characteristically in the cytoplasms of oligodendrocytes of brains with multiple system atrophy. We examined whether proline-directed protein kinases, which have been found in several neuronal inclusion bodies such as neurofibrillary tangles and Lewy bodies, are associated with GCI. We unexpectedly have observed cdk 5 and MAPK in GCI. These kinases were not immunolabeled in coiled bodies which are oligodendroglial inclusion bodies in brains with progressive supsranuclear palsy and Alzheimer's disease. We also found microtubule-associated protein 2 in oligodendrocytes in brains with MSA, which have not been observed in normal controls or neurological disease controls. Cdk 5 and MAP 2 are principally neuronal proteins. MAPK have been found in neuronal somata and some astrocytes, but not in oligodendrocytes. Thus, the present results suggest that oligodendrocytes in MSA harbor an abnormal phenotypic nature in terms of the aberrant expressions of the principally neuronal proteins. PMID- 10377788 TI - [Campylobacter jejuni enteritis and Guillain-Barre syndrome]. AB - Guillain-Barre syndrome (GBS) is the most common cause of acute neuromuscular paralysis. Sera from patients with GBS following Campylobacter jejuni infection frequently have autoantibody to GM 1 ganglioside in the acute phase of the illness. We revealed that the lipopolysaccharide (LPS) of C. jejuni that was isolated from a GBS patient has the oligosaccharide structure [Gal beta 1-3 GalNAc beta 1-4 (NeuAc alpha 2-3) Gal beta 1-], which is identical to the terminal tetrasaccharide of GM 1 ganglioside. (1) Infection by C. jejuni that bears the GM 1-like lipopolysaccharide associated with the serotypic determinant of PEN 19 induces high production of IgG 1 and IgG 3 anti-GM 1 antibodies with help of T cells. (2) IgG anti-GM 1 antibody binds to motor nerve terminal axons, inhibits motoneuron excitability, and produces the development of GBS. PMID- 10377789 TI - [DPB 1 *0501-associated opticomyelitis and atopic myelitis: novel disease entities]. AB - To clarify the relationship between Th 1/Th 2 balance and clinical features, we studied the intracellular IFN gamma-positive versus IL-4-positive cell ratio in peripheral blood CD 4 T cells by flow cytometry and measured total and allergen specific IgE by ELISA in 227 patients with various neurologic diseases including multiple sclerosis (MS), myelitis and HAM/TSP, and 42 healthy hospital subjects. The intracellular IFN gamma/IL-4 ratio in the patients with acute myelitis was significantly decreased, and the total serum IgE level and frequency of mite antigen-specific IgE were significantly elevated as compared with the controls. Patients with HAM/TSP, however, had a significantly higher intracellular IFN gamma/IL-4 ratio, lower total IgE level, and lower frequency of cedar pollen specific IgE than did the controls. The patients with recurrent opticomyelitis (ROM) had a significantly higher frequency of relapse, higher EDSS score, and lower number of brain MRI lesions than the patients with conventional MS, and only those with ROM showed a significant association with the HLA-DPB 1 *0501 allele. The ROM patients had a significantly higher intracellular IFN gamma/IL-4 ratio and IL-4-/IFN-gamma + cell percentages than the controls. These findings suggest that the Th 2 cell response predominates in acute myelitis with hyperIgEaemia (atopic myelitis), whereas the Th 1 cell response predominates in ROM and HAM/TSP. PMID- 10377790 TI - [Pathogenetic significance of HTLV-I infection and immune surveillance in HAM]. AB - HTLV-I proviral DNA load is significantly increased in HTLV-I associated myelopathy (HAM) compared with asymptomatic HTLV-I seropositive carriers (SPC), and this spread of HTLV-I infection seems to be critically important in the pathogenesis of HAM. Thus, in this report, cellular immune surveillance against HTLV-I was reviewed. (1) MHC class I-restricted cytotoxic T lymphocytes (CTL) activities are detected in peripheral blood mononuclear cells (PBMC) of HAM. CTL release various proinflammatory and cytotoxic cytokines, chemokines, and proteases. Since CTL are also found in the spinal lesions of HAM, CTL may contribute to the tissue damage. In spontaneous proliferation of PBMC in HAM, CD 4/CD 8 is decreased due to the proliferation of CD 8 + CTL. CD 4/CD 8 is inversely correlated with the clinical severity of HAM. Collectively, CTL may be involved in the pathogenesis of HAM. (2) Activity and subsets of natural killer (NK) cells are lower in HTLV-I-seropositive individuals. Moreover, NK have only a weak cytotoxicity against HTLV-I infected cells. (3) Antibody-dependent cell mediated cytotoxicity (ADCC) are impaired in HAM compared with SPC due to the suppressed effector cell activity. Since ADCC effectively lyse HTLV-I infected cells in vitro, the impaired ADCC may in part allow the spread of HTLV-I infection in HAM, and potentiation of ADCC may have an anti-HTLV-I therapeutic effect. PMID- 10377791 TI - [Infection and multiple sclerosis]. AB - Multiple sclerosis is a putative autoimmune disease in which limited numbers of autoimmune T cells reactive to myelin basic protein or proteolipid protein would play critical roles in initiation or augmentation of the inflammatory processes. Owing to the recent immunological studies, it is now possible to discuss the possible link between infectious agents and the development of MS on the molecular terms. This article deals with key issues in this subject such as molecular mimicry between autoantigen and viral peptide, autoimmune T cell stimulation by bacterial superantigens, and regulatory network in MS. In addition to reviewing the current activity in other laboratories, I point out that there might be much more broader range of peptide ligands for autodestructive T cells causing MS. This postulate is based on our recent discovery for the presence of degenerate autoimmune T cells in animal model of MS (EAE). I also indicate the possibility that immune regulatory system could be destroyed by some infectious agents. These informations should have significant implications for management of MS patients. PMID- 10377792 TI - [Infection in the central nervous system and corticosteroid therapy]. AB - No standardized therapy has been established for viral encephalitis except for herpes simplex encephalitis. Not a few neurologists, however, have had an impression that administration of corticosteroids ameliorated neurological impairment and induced better prognosis in some patients with viral encephalitis. Five patients with aseptic meningitis and 9 patients with viral encephalitis, who were moderately to severely ill, were examined for cerebrospinal fluid (CSF) parameters before and after short-term intravenous administration of corticosteroids. In all of the patients with aseptic meningitis, severe headache and nausea disappeared rapidly with this treatment, which was accomplished via anti-inflammatory effects of corticosteroids. By contrast, 5 of the patients with viral encephalitis responded well to intravenous corticosteroids, whereas the remaining 4 patients did not, three of whom showed poor prognosis. The CSF containing more than 15% of CD4+CD26+ memory helper T cells guaranteed good response to corticosteroid therapy. It appeared that viral encephalitis with severe inflammation had poor prognosis irrespective of attempted therapy including intravenous corticosteroids. The CSF of patients who responded well to corticosteroid treatment showed a significant reduction in CD4+CD29+ helper inducer T cells in the course of the illness. This finding indicates that autoimmune mechanisms may be involved in the pathogenesis of neurological impairment in a part of patients with viral encephalitis. PMID- 10377793 TI - [Functional imaging for disorders of basal ganglia]. AB - The nigrostriatal dopaminergic function and regional glucose metabolism were evaluated in patients suffering from various disorders of basal ganglia by using positron emission tomography with 18F-dopa and 18F-FDG, respectively. The 18F dopa uptake in the striatum (the caudate head and the putamen) decreased in patients with Parkinson's disease but was relatively unaffected in the caudate. The cerebral glucose metabolism was normal in patients with Parkinson's disease. The 18F-dopa uptake in the striatum also decreased in cases of multiple system atrophy and progressive supranuclear palsy, but there was no difference in the uptake between the caudate and the putamen. The glucose metabolism decreased in the cerebral cortices and the striatum: this finding was also different from those of Parkinson's disease. A normal 18F-dopa uptake with a markedly decreased striatal glucose metabolism was observed in cases of Huntington's disease. The 18F-dopa uptake increased and the glucose metabolism was normal in cases of idiopathic dystonia. Various patterns of 18F-dopa uptake and glucose metabolism were thus observed in the various disorders of basal ganglia. These results suggest that the measurements of the 18F-dopa uptake and glucose metabolism would be useful for evaluating the function of the basal ganglia in various disorders of basal ganglia. PMID- 10377794 TI - [PET study of cholinergic system in the brain]. AB - Recently, we have developed a method to measure acetylcholinesterase (AChE) activity, a functional marker for cholinergic system, by positron emission tomography (PET) and carbon-11 labeled N-methyl-4-piperidyl acetate. Kinetic analysis of the radioactivity in the brain and the plasma yielded a rate constant "k 3" as an index of AChE activity. The ratios for the k 3 values for the cerebral cortex/thalamus/cerebellum/striatum found in healthy participants were 1/3/8/10, respectively, corresponding well with AChE activity ratios in the brain at necropsy (1/3/8/38), except for the striatum. In 23 healthy volunteers (age range: 24-89 years), there was no age-related decline of k 3 values in the cerebral cortex, suggesting AChE activity is preserved in aged cerebral cortex. In 11 patients with Alzheimer's disease, there was a significant reduction (-24%) of k 3 values in the cerebral cortex and hippocampus, suggesting a loss of ascending cholinergic system from the basal forebrain to the cerebral cortex and hippocampus. In 16 patients with Parkinson's disease, there was a significant reduction (-18%) of k 3 values in the cerebral cortex. In 10 patients with progressive supranuclear palsy, there was a significant reduction (-38%) of k 3 values in the thalamus. This technique is useful for investigating central cholinergic system in neurodegenerative disorders with dementia. PMID- 10377795 TI - [SPM analysis on PET and SPECT data]. AB - I described the methodological explanation of statistical parametric mapping (SPM) with the special attention to the clinical applications. SPM was designed mainly by K. Friston and R. Frackowiack to visualize the statistically significant regions on the CBF data sets obtained from PET or SPECT during a various kinds of activation. In short, images were realigned to the first scan to eliminate the head position movements, and were normalized to the standard brain shape (Talairach & Tournoux) by linear or non-linear transformation. After that, gausian filtering was done in order to eliminate the individual brain differences and to increase the signal to noise ratio. The pixel by pixel analysis was done with multiple comparison correction, and the statistically significant areas were displayed. In clinical applications, comparison of normal controls with Alzheimer's disease, diffuse Lewy body disease, and some activation study in the pathological vs normal condition. In SPECT, we can obtain the brain perfusion images by the injection of Tc-99m labeled CBF tracers, so that we can analyze the condition with the subjects not confined to the scanner. PMID- 10377796 TI - [BOLD functional MRI: practical pitfalls]. AB - There is mounting criticism regarding the validity of blood oxygenation level dependent contrast functional magnetic resonance imaging (BOLD-fMRI) studies. The main sources of error in BOLD-fMRI are pixel misalignment and physiological noise. One of the widely practiced tactics to ameliorate motion related fictitious activation is post-processing motion correction algorithms. Nevertheless, the validity of activation data following application of such algorithms has not been totally confirmed. In this article, the practical pitfalls of BOLD-fMRI and potential correction algorithms are concisely discussed. PMID- 10377797 TI - [Functional localization of the somatomotor area by magnetoencephalography]. AB - Precise localization of the current dipole by the magnetoencephalography (MEG) has enabled us to combine the functional information onto the anatomical landmarks. This merit can be best exhibited when the dipole is situated in the superficial cortex and directed parallel to the skull surface. However, before utilizing MEG extensively as a clinical tool, it is inevitable to confirm the precision of the source localization by comparing the estimation with the actual sources. Somatosensory evoked field (SEF) following the electric shock to the peripheral nerve and movement-related cortical field (MRCF) associated with self paced movement can show us the estimated sources at the postcentral and precentral cortex, respectively, with somatotopic organization. These localizations were confirmed by the direct recordings from the human brain surface during the operation, even if the corresponding areas were anatomically distorted by some lesion occupying the central area. In addition, MEG can localize second somatosensory area (SII) over the superior bank of the Sylvian fissure as well as posterior parietal cortex (PPC), which are difficult to be detected by the EEG recording. These reliable estimation enables us to apply MEG to clarification of pathogenesis of various diseases and source localization for higher brain function. PMID- 10377798 TI - [Clinical characteristics of senile dementia in Japan]. AB - Senile dementia encountered in Japan is characterized with a higher frequency of vascular dementia as compared to that in North America and Europe. This clinical impression is well substantiated by the statistics from the epidemiologically established cities and townships such as Rochester, Minnesota and Framingham, Massachusetts in the United States and Hisayama, Fukuoka in Japan. However, the difference in incidence between vascular dementia and Alzheimer disease in Japan has narrowed down, and it is possible that the ratio between those two disease entities in Japan may approach that in North America and Europe in the future. In addition, there are qualitative differences in vascular dementia in Japan in that the most frequent vascular dementia in Japan is caused by multiple lacunar infarcts and that one half of those patients develop dementia insidiously without a single ischemic event clinically. Those characteristics make the use of some of commonly utilized diagnostic criteria for vascular dementia inapplicable, and make it necessary to develop a comprehensive and flexible criterion for vascular dementia which is universally applicable in the world. PMID- 10377799 TI - [Prevalence, incidence, and risk factors of vascular dementia: the Hisayama study]. AB - We studied the type-specific prevalence, incidence, and risk factors of vascular dementia in elderly persons from a Japanese community of Hisayama. In 1985, we performed a screening survey of dementia among 887 Hisayama residents > or = 65 years or older (screening rate, 95%), using clinical information and Hasegawa's dementia scale, and consequently, determined 59 subjects as demented. Of these, 58 cases underwent brain examinations at autopsy and/or computed tomography during the subsequent 12.5 years. Among the 58 cases of dementia, the frequency of vascular dementia (VD) was 43%: the rate was 2 times higher than that for Alzheimer's disease (AD). In the subjects of VD, the most frequent type of stroke was due to small-artery disease, which caused multiple lacunar infarction (40%) and Binswanger's disease (12%). We also followed the 826 nondemented subjects for 7 years starting in 1985 in order to determine the type-specific incidence of dementia and its risk factors in the general population. The age-adjusted total incidence (per 1,000 person-years) of dementia was 19.3 for men and 20.9 for women. The corresponding rates of VD were 12.2 for men and 9.0 for women, and for AD 5.1 for men and 10.9 for women. Among the VD subjects whose brain morphology was examined, the most frequent type of stroke was multiple lacunar infarcts (42%), but half these subjects lacked a stroke episode in their histories. Multivariate analysis showed that age, prior stroke episodes, systolic blood pressure, and alcohol consumption were significant risk factors for the occurrence of VD. PMID- 10377800 TI - [Young-adult-onset hereditary subcortical vascular dementia: cerebral autosomal recessive arteriosclerosis with subcortical infarcts and leukoencephalopathy (CARASIL)]. AB - The clinical features of a probably autosomal recessive syndrome ("CARASIL"), yet to be confined in Japan and characterized by prematurity of vascular dementia, alopecia and spondylosis deformans are reviewed through comparison with cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), which has been reported in Europe and North America, and recently in Japan. These two syndromes have many common features, such as familiality, encephalopathy of Binswanger type, and absence of vascular risk factors. There exists, however, a number of differences as follows: (1) Onset of encephalopathy is 32 years of age in "CARASIL" vs. 45 in CADASIL. (2) Male to female ratio is 3.2: 1 vs. 2:1.(3) Two thirds of "CARASIL" patients show stroke and/or stepwise deterioration, while almost all CADASIL patients have stroke. (4) Associated psychiatric features are euphoria, emotional lability and loss of spontaneity vs. severe mood disorders. (5) Migraine is a cardinal feature of CADASIL and vasospasm may occur during cerebral angiography. (6) White matter lesions on MRI are diffuse and homogeneous vs. punctuated and nodular. The latter four differences may mirror the difference in the pathology of arteriopathies. "CARASIL" is clearly different from CADASIL and reflect a second genetic condition with a seemingly direct effect upon the cerebral vasculature. PMID- 10377801 TI - [Identification of Notch 3 mutation in the first Japanese CADASIL family]. PMID- 10377802 TI - [A clinicopathologic study of progressive subcortical vascular encephalopathy of the Binswanger type (PSVE)]. AB - From an electron microscopic study I indicated that the number of nerve fibres per unit area in the frontal white matter was significantly less in PSVE than that in controls. The white matter pallor in PSVE is mainly based on the loss of nerve fibres. The dementia in PSVE is probably related to the loss of nerve fibres in the cerebral white matter. The extent of frontal white matter pallor tended to be less broad in vascular parkinsonism (VP) than in PSVE without parkinsonism. The relatively slight damage of the white matter may contribute to VP observed in PSVE. The number of oligodendrocytes and of astrocytes was decreased in PSVE showing half of that in the deep white matter of the controls. This suggests that the loss of oligodendrocytes and astrocytes can play a role to the process of nerve fibres loss in PSVE. Not only the severe hyalinotic changes in the white matter arterioles, but also the diffuse atherosclerotic lesions in the main stem of cerebral arteries were the characteristic arterial lesions in PSVE. On the basis of these arteriosclerotic lesions, we speculated that the repeated hypotension or the greater variability of casual blood pressure could induce PSVE in the elderly hypertensives. However, the retrospective analysis of casual blood pressure did not support our hypothesis. PMID- 10377803 TI - [The role of immunologic reactions in the pathogenesis in Binswanger's disease; a clue to therapeutic approach]. AB - We examined the alterations of glial cells in the brains with Binswanger's disease. In comparison to the brains with lacunar cerebral infarction and those from non-neurological controls, oligodendroglia was decreased in number, and microglia was increased and activated in the white matter lesions in Binswanger's disease. Astroglia occasionally showed a regressive change, termed clasmatodendrosis. In rodents, similar white matter lesions could be induced after chronic cerebral hypoperfusion along with a numerical decrease of oligodendroglia, an increase of astroglia and activation of microglia in the corresponding white matter. These white matter lesions were suppressed by injection of immunosuppressant, cyclosporin A or FK506, with a suppression of microglial activation. These findings indicate that the lesion development in the white matter may be mediated by an immunologic reaction. Unfortunately, these drugs may be inappropriate for a practical use in terms of their side effects. Alternatively, we tried the use of anti-thrombin drug, argatroban. In acute neurological exacerbation of Binswanger's disease, markers for coagulation fibrinolysis pathway were elevated significantly, and this activation was suppressed along with a recovery of neurological symptoms such as parkinsonism and pseudobulbar palsy. Thrombin contributes to a microcirculatory disturbance and may also enhance an inflammatory response through the action of thrombin receptor. PMID- 10377804 TI - [Blood pressure change abnormalities on lacunar infarcts and ischemic white matter lesions]. PMID- 10377805 TI - [Abnormal hyperexcitability in ALS]. AB - The defect of neuromuscular transmission is one of the important signs in ALS. The amplitude of a single motor unit potential from patients with ALS often decrease during tonic voluntary contraction. This phenomenon is closely correlated with fatigue seen in the patient. Overfunctioning of Ach release in the nerve terminal might cause the failure of neuromuscular transmission in ALS. Fasciculations is an another characteristic sign and considered mainly to be peripheral axons in origin. It is postulated that the dysfunction of potassium channel in ALS axons makes the hyperexcitability of the axon membrane, causing fasciculations. Magnetic cortical stimulation sometimes evokes the same potentials as fasciculations, implying the hyperexcitability might be present also in spinal motoneurons or even in pyramidal neurons in ALS. All of these findings lead to the hypothesis that hyperexcitability or overactivity of central and peripheral motoneurons is an essential feature in ALS. PMID- 10377807 TI - [Approach to study of etiology and therapy of ALS using the animal models]. PMID- 10377806 TI - [Neuropathology of the motor neuron disease--Bunina body]. AB - Bunina body is known to occur in cases of sporadic amyotrophic lateral sclerosis (ALS), ALS with dementia (a so-called Mitsuyama type), and Guamanian ALS, and seems to lend a diagnostic priority to the presence of Bunina bodies. Absence of Bunina body in a subset of familial ALS with posterior column and spinocerebellar tract involvement or motor neuron disease with basophilic inclusion also adds credence to the specificity of Bunina body in ALS. However, despite its bright eosinophilia, distinct expression of cystatin C, and conspicuous ultrastructure, the origin of Bunina body is still unknown and remain several unsolved problems. Bunina bodies are usually seen within the cytoplasm or dendrites of degenerated and/or sometimes normal-looking large neurons. However, so far, no Bunina body has been found within the axoplasm. Bunina bodies are mainly distributed in the lower motor neurons. Only a single report described it in the Betz cell. Furthermore, several recent studies have revealed the occurrence in neurons which are so far considered to be exempt from the pathology of ALS, i.e. the oculomotor nucleus, Onufurowicz's nucleus, Clarke's nucleus, reticular formation of the brain stem, and subthalamic nucleus. In addition to the occurrence in neurons other than motor neurons, ultrastructurally similar or identical inclusions are reported in neurons of aged rats, the olfactory bulb of aged human, the spinal cord of a patient without ALS, and gangliocytoma. Bunina body probably represents a facet of the degenerating process of a neuron. A high incidence in ALS, particularly in lower motor neurons, awaits a further study of the pathogenesis of Bunina body. PMID- 10377808 TI - [Molecular mechanism of ALS and a possible gene therapy]. AB - We report clinical characteristics of familial amyotrophic lateral sclerosis (FALS) with four different missense point mutations in exons 1, 2, 4, and 5 of the Cu/Zn superoxide dismutase (SOD) gene, that result in amino acid substitutions of cysteine 6 by phenylalanin (C 6 F), histidine 46 by arginine (H46R), leucine 84 by valine (L84V), isoleucine 104 by phenylalanine (I104F), and valine 148 by isoleucine (V148I), in five Japanese families. Although features of progressive neurogenic muscular atrophy was common in patients of these families, patients of each family showed characteristic clinical features. Immunoreactivity for Cu/Zn SOD of the motor neurons was not different between the ALS and controls. In contrast, immunoreactivity for NT was densely detected in motor neurons of ALS while that was not or was only minimally detected in those of controls. Adenovirus-mediated E. coli LacZ gene was transferred and expressed both in the muscle and spinal cord of transgenic mice. These results suggest that familial ALS with different mutations of the Cu/Zn SOD gene showed each clinical characteristics, that nitration of protein-tyrosine residue is upregulated in motor neurons of the spinal cord of ALS, and that there could be a possible future therapy of ALS with exogenous gene transfer. PMID- 10377809 TI - [Mechanical ventilation for amyotrophic lateral sclerosis--Making a comparison between hospital and home care]. AB - In Japan, the traditional method of mechanical ventilation (MV) used for patients with amyotrophic lateral sclerosis (ALS) has been positive pressure ventilation with tracheostomy (PPV). Survival after starting PPV can be for many years. The purpose of our study is to report our findings on 25 patients (22.5%) using PPV for thirteen years. A total of 91 ALS patients without PPV died during in past thirteen years. Four patients with PPV lived at home and their families usually served as primary caregivers. Other five patients with PPV lived in our hospital, because they were lacked willing, competent or available caregivers at home, and they had not sufficient informed consent about use of mechanical ventilation or not. The quality of MV care provided by properly trained family members and home helper was as good as care provided by home-visited nurse. Quality of life in ALS patients with home MV were significantly increased comparing to hospital MV. Few patients chose nasal intermittent PPV. Most patients with ALS do not want to use MV. They need assistance with palliative care and planning for emergencies, so that PPV can be avoided when respiratory failure occurs. Before a decision is made to use PPV, the patient with ALS and family should understand that death related to ALS can be prevented with home MV and good care. However, if PPV is used, ongoing tracheal suctioning will be required, immobility will progress, or needed resources and cost are high, with the result that family might be burdened with heavy responsibility. Thorough informed consent are necessary. PMID- 10377810 TI - [Surgical treatment for intractable epilepsy: update and future]. AB - For successful surgical treatment of intractable epilepsy, identification of the epileptogenic area and functional cortex, by using the intracranial electrodes such as subdural and depth electrodes, is important. Since 1994, via chronic subdural electrodes recording, we performed anterior temporal lobectomy with hippocampectomy for 18 patients with temporal lobe epilepsy. For 10 patients with extratemporal lobe epilepsy, cortical resection of the epileptogenic cortex was performed. For the epileptogenic cortex overlapping with functional area, we added the multiple subpial transection. Favorable postoperative seizure outcome was obtained in most of the patients. Although non-invasive presurgical evaluation modalities such as MRI, video-EEG monitoring, MEG, and FDG-PET are useful in the diagnosis of epilepsy, it is impossible to localize precisely the exact epileptogenic zone and functional cortex. PMID- 10377811 TI - [General principles of treatment and effects of childhood intractable epilepsy]. AB - Intractable epilepsy can be defined as (1) refractory to the treatment, (2) with frequent and severe seizures, and (3) with association of mental disability. Main disorders of childhood intractable epilepsy include Ohtahara syndrome, West syndrome, Lennox-Gastaut syndrome, severe myoclonic epilepsy in infancy, and symptomatic localization related epilepsy. Our results of long-term prognosis of West syndrome showed seizure free patients were 11 of 14 (79%) in cryptogenic and 10 of 29 (34%) in symptomatic cases. Intelligence was normal in 9 of cryptogenic (64%) and only 1 of symptomatic cases. Evolutional changes of West syndrome showed seizure free in 21, localization related epilepsy in 10, secondary generalized epilepsy in 10, unclassified epilepsy in 2. None of Lennox-Gastaut type was observed. A 13-year-old boy of tuberous sclerosis with frequent drop attacks was treated by anterior 3/4 callosotomy. Although the tonic seizures remained, drop attacks were completely abolished. The measures to the intractable epilepsy should be targeted to the symptoms or conditions to which the patients and guardians want to be eliminated. PMID- 10377812 TI - [Current status and future developments in the treatment of medically intractable epilepsy in adults]. PMID- 10377813 TI - [Electrophysiological study of intractable epileptogenicity in human epilepsy: clinical usefulness of ictal DC shifts and cavernous sinus EEG]. AB - In order to clarify the clinical and electrophysiological features in intractable epileptogenicity in human epilepsy, we applied the new techniques, ictal DC shifts and cavernous sinus EEG recording, for presurgical evaluation of patients with intractable partial epilepsy. (1) Ictal DC shifts were successfully recorded with subdural electrodes in 8 patients with intractable neocortical epilepsy, and an analysis of ictal DC shifts would add useful information to delineate an epileptogenic area. Scalp-recorded ictal DC shifts were also investigated in 3 patients with intractable neocortical epilepsy. It also delineated the epileptogenic area, but it was vulnerable for artifacts. (2) By using the techniques of intravascular EEG recording, we recorded EEG from the bilateral cavernous sinus (cavernous sinus EEG) in patients with intractable temporal lobe epilepsy. Cavernous sinus EEG well sensitively recorded interictal, also ictal in selected patients, epileptiform discharges which arose from the mesial temporal structure even though they were not recorded by scalp electrodes. It is concluded that the above two techniques are clinically useful for delineating an epileptogenic area in patients with neocortical epilepsy and temporal lobe epilepsy. PMID- 10377814 TI - [Psychiatric disorders following and preceding temporal lobectomy]. AB - We analysed pre- as well as post-operative psychiatric disorders in thirty eight patients with temporal lobe epilepsy. While postoperative paranoid disorders were closely correlated with preoperative acute interictal psychoses, episodes of postictal psychoses prior to surgery were associated with postoperative mood disorders. A good prognosis of postoperative mood disorders was stressed. The literature search supported the predominance of right-sided lobectomy in patients with de novo psychoses after surgery. The relationship between left-sided lobectomy and postoperative mood disorders needs further amplification and remains tentative. We stressed the need for a prophylactic psychotherapy to surgical candidates to cope with unrealistic wishes to get relieved from all the difficulties in their lives after temporal lobectomy. PMID- 10377815 TI - [Increased vascular endothelial growth factor (VEGF) is causative in Crow-Fukase syndrome]. AB - Crow-Fukase syndrome is a rare multisystem disorder characterized by polyneuropathy, organomegaly, endocrinopathy, skin changes and M-protein. We have examined levels of vascular endothelial growth factor/vascular permeability factor (VEGF) in serum with Crow-Fukase patients. Serum VEGF levels in Crow Fukase syndrome were about 15 to 30 times higher than control and other neurological disorders. Most of the characteristic manifestations may be well explainable by the biological function of VEGF except polyneuropathy. We examined the direct effects of VEGF on blood nerve barrier function using blood brain barrier model of rat and intraneural injection of recombinant VEGF. As the result, VEGF affected blood nerve barrier and increased microvascular permeability, thereby inducing endoneurial edema. After increasing the permeability of the blood nerve barrier by VEGF, serum components toxic to nerves such as complements and thrombins may induce nerve damage. Our results suggest that overproduction of VEGF plays an important role in the pathogenesis of Crow Fukase syndrome. PMID- 10377816 TI - [Ischemic neuropathy]. PMID- 10377817 TI - [New trend in pathogenesis of diabetic neuropathy]. AB - The pathogenesis of diabetic neuropathy remains unclear, although several factors have been implicated in its pathogenesis. We have examined possible roles of decreased production of nitric oxide, ion channel dysfunction and decreased capacity of nerve regeneration. STZ-induced diabetic rats showed decreases in nociceptive threshold and NADPH-diaphorase positive neurons, nNOS level and cGMP content of DRG at 12 weeks after induction of diabetes. The rats injected by L NAME, potent nNOS inhibitor, showed decreased nociceptive threshold, although D NAME, inactive in nNOS inhibition, did not. These results suggest that decreased NO production might be involved in hyperalgesia in diabetic rats. Both hyperglycemia and decreased Na/K-ATPase activity are thought to be characteristic features of diabetic neuropathy. To investigate the presence of ion channel abnormality in diabetic nerves, a Vaseline-gap voltage clamp technique was applied for a single myelinated fibers under 30 mM high glucose plus 0.1 mM ouabain. Since K current was increased, a Ca activated K channel blocker was applied and this increase was shown to be suppressed. Furthermore, Ca channel blockers all suppressed increased K currents, suggesting that the condition induced an increase of Ca influx, thereby increasing Ca activated K currents through K channels. The data are important in that diabetic condition may induce both Ca influx, leading to nerve degeneration, and increased K current, resulting in decreased nerve conduction. Nerve regeneration has been known to be disturbed in diabetic condition. We have shown a decrease in nerve elongation rate in diabetic rats after crush of sciatic nerve, although this decrease was not ameliorated by ARI. Furthermore, Wallerian degeneration was shown to be delayed in diabetic nerves, leading to delayed nerve regeneration. Hyperphosphorylation of both medium and high molecular weight neurofilaments that might be induced by protein kinases including CDK 5 may be involved in the mechanism. PMID- 10377818 TI - [Immunopathology of inflammatory neuropathies]. AB - With the use of immunohistochemical technique, nerve biopsy is more informative for the diagnosis of inflammatory neuropathies. In chronic inflammatory demyelinating neuropathy, an increased number of T cells are frequently present in endoneurium, which is in contrast to hereditary neuropathies. In active demyelinating lesions, macrophages adhering nerve fibers showed stainings with TNF-alpha. NOS and cyclooxygenase-2 (COX-2). These molecules may act in concert to promote nerve damage. The inhibitor of COX-2, nimesulide, was effective on experimental allergic neuritis, even if given after the onset of clinical signs. A COX-2 inhibitor may have potential as an additional therapeutic agent in human inflammatory neuropathies. In vasculitic neuropathies, cell-mediated cytotoxicity may be involved in the pathogenesis of small vessel injury. Axonal injury may be caused by focal ischemia. However, an immune attack might be involved in nerve damage, since T cells and IL-12 positive cells were found in endoneurium of some patients with active vasculitis. PMID- 10377820 TI - [Antiganglioside antibodies in the pathogenesis of autoimmune neuropathies]. AB - Antiganglioside antibodies are frequently detected in sera from patients with autoimmune neuropathies, such as Guillain-Barre syndrome, Miller Fisher syndrome, IgM paraproteinemic neuropathy, chronic inflammatory demyelinating polyneuropathy, and multifocal motor neuropathy. In the acute phase sera from GBS patients, antiganglioside antibodies are detected in 60-70%. Ganglioside antigens recognized by serum antibodies are varied from case to case. IgG antibody against GQ1b ganglioside is specifically raised in sera from patients with Miller Fisher syndrome and Guillain-Barre syndrome with ophthalmoplegia. That antibody may bind to the paranodal myelin of oculomotor, trochlear and abducens nerves, where GQ1b ganglioside is specifically localized, to cause ophthalmoplegia. IgM M-protein which recognizes the disialosyl residue of GD1b is specifically associated with sensory ataxic neuropathy. The IgM M-protein may bind to the primary sensory neurons, where GD1b ganglioside is localized, to cause sensory disturbance. After we confirmed the localization of GD1b in the rabbit primary sensory neurons, we sensitized rabbits with GD1b and induced sensory ataxic neuropathy in them. This is the first established animal model of autoimmune neuropathy induced by sensitization with ganglioside. Some antiganglioside antibodies may determine the clinical phenotype of neuropathy by binding specifically to the ganglioside antigens which have unique localization. PMID- 10377819 TI - [Expression of GDNF and GDNFR-alpha mRNAs in human peripheral neuropathies]. PMID- 10377821 TI - [Anti-GM1 antibodies did not induce conduction block in vivo]. PMID- 10377822 TI - [Differences in the properties of sensory and motor axons]. PMID- 10377823 TI - [Clinical neurophysiological evaluation of the motor unit]. PMID- 10377824 TI - [Electrophysiological studies of the corticomotoneuron in ALS]. PMID- 10377825 TI - [Physiological studies of the motor cortex]. PMID- 10377826 TI - [Cortical mechanisms underlying sensori-motor association and its disorders]. PMID- 10377827 TI - [Neuroimaging in patients with CVD]. AB - The development of the neuroimaging technique has revolutionized clinical neurology. This is also true in the field of stroke. Because of the easy accessibility and little time consuming, the computed tomography (CT) should be applied first to the stroke patient and other examinations should be prepared later to the patient due to the severity and the characteristic of the lesion. These are magnetic resonance imaging (MRI) (including MR angiography. MR spectroscopy, diffusion MRI), cerebral angiography, neurosonography (carotid ultrasound, color flow imaging, transcranial Doppler), single photon emission computed tomography (SPECT) and positron emission tomography (PET), etc. It is a matter of common sense that CT and MRI are now particularly important among laboratory examinations for neurological diseases and these non-invasive techniques will become more important for the elderly stroke patients. Recent study cleared the combination of 3-dimensional computed angiography and neurosonography has an informative value and this is especially important for the elderly patients who are fragile and need rapid and accurate diagnosis. PMID- 10377828 TI - [Current diagnosis and treatment of Parkinson's disease]. PMID- 10377829 TI - [Diagnosis and treatment of multifocal motor neuropathy (Lewis-Sumner)]. AB - We made a retrospective long-term follow-up study of 25 patients with multifocal motor neuropathy (Lewis-Sumner). The diagnosis was based upon criteria modified from those of AAEM (Sumner 1997). The electrophysiological findings indicating conduction block or focal demyelinative lesions were more diagnostic than anti-GM 1 antibody titers, which were elevated in only 40% of these patients. Demonstration of definite conduction block was not always possible in those patients who responded favorably to intravenous immunoglobulins (IVIg), whereas indirect pieces of evidence such as F-wave abnormalities or focal conduction delay or dispersion were equally helpful. IVIg had superior outcome to cyclophosphamide, which sometimes caused serious adverse effects. Three patients with severe axonal involvement showed elevated monospecific antibodies to GalNAc GD1a. PMID- 10377830 TI - [Neuroradiologic and pathologic approaches to the diagnosis of dementia syndrome]. AB - "Dementia" is the general term used to describe the symptom complex of intellectual deterioration in adult. Interest in accurately diagnosing dementia is a relatively recent phenomenon. This is reflected in both the development of neuroradiologic examinations, including MRI and SPECT as well as PET, and marked increase in both the incidence and prevalence of dementia associated with increase of the elderly population. The clinical evaluation remains the key to the differential diagnosis. Most cases of "typical dementia" can be diagnosed accurately by clinical criteria. However, the definitive diagnosis of "atypical dementia" still requires intensive neuroradiologic studies and histologic examination of brain to identify characteristic structural changes. In this study, we presented both neuroradiologic and neuropathologic information, which is important in diagnosing diseases that present atypical dementia syndrome. These diseases are as follows; AIDS, isolated CNS angiitis, CO intoxication. Wernicke encephalopathy, adrenoleukodystrophy, Nasu disease, CADASIL, CARASIL, glioblastoma, primary CNS lymphoma, antiphospholipid antibody syndrome, reversible posterior leukoencephalopathy syndrome, mitochondrial encephalopathy (MELAS), and subcortical vascular dementias. PMID- 10377831 TI - [Evoked potential]. PMID- 10377832 TI - [Magnetic resonance imaging in multiple sclerosis]. AB - Useful characteristics of MRI finding of multiple sclerosis (MS) include the distribution of lesions such as a strictly periventricular, infratentorial, or juxtacortical location, involvement of the corpus callosum and the presence of ovoid lesions with long axis directed to lateral ventricles. Our MRI-diagnostic criteria improved the sensitivity and specificity and is clinically useful. New sequences, such as fast spin echo, turbo spin echo or fluid attenuated inversion recovery have improved the detection of lesions. The presence of contrast enhancement in some but not all lesions--that is, evidence of both old and new lesions--provides additional diagnostic support. Enhanced lesions with more than 1 cm diameter often become ring-shaped. Strong correlations were found between the number and volume of enhancing lesions with changes of T 2 and magnetization transfer (MT) lesion loads in patients with secondary progressive MS. The degree of hypointensity of so called black holes on moderately T1-weighted spin echo images correlates with loss of magnetisation transfer, a marker of destruction of matrix and axon, and shows correlation with disability. One of spectroscopic indices of axonal loss is N-acetylaspartate. Atrophy is a process closely linked with the progressive phase of MS and worsening disability. Detection of the reduction in cord cross-sectional area or spinal cord atrophy over time makes an important contribution to the evaluation of therapeutic efficacy, especially in primary progressive disease. PMID- 10377833 TI - [Laboratory identification of blood hypercoagulability]. AB - For many years, the laboratory investigation of patients with thrombophilia has lagged behind that of patients with bleeding diathesis. The improved understanding of the mechanisms that control and regulate coagulation, and resultant recognition of new defects have greatly stimulated clinical laboratory interest in this area. Assays regarding the developed resistance to activated protein C, deficiencies of antithrombin, protein C and protein S, and the presence of antiphospholipid antibodies are widely available and should be a part of investigations of patients with idiopathic thrombosis. Such a study would likely provide an explanation of thrombosis in 40-60% of patients. Abnormalities of fibrinogen and fibrinolysis may be explained, although such defects are currently considered rare. More sophistic assays are being developed to detect abnormalities de to factor V Leiden and prothrombin 20,210 gene mutation, which will undoubtedly detect more patients with thrombophilia. Laboratory tests to define the hypercoagulable state are continually being developed. They include tests for novel activation markers. However, acceptance of these approaches by clinical laboratories has been slow. PMID- 10377835 TI - [The effect of sex factors on cytologic changes in the sputum of young adults exposed to urban air pollution]. AB - The harmful effect of occupational and urban air pollution on the function and structure of the respiratory tract has been proved. The severity and incidence of epithelial changes are in direct relation to duration of exposure and age of the examined population groups [1]. Squamous metaplasia of bronchial epithelium is an indicator of predisposition to chronic obstructive pulmonary disease and lung cancer [1-5]. Due to its topography and the development of industry and traffic, Sarajevo used to be the city with an extremely high level of air pollution [10 12]. Some studies in adults showed that the respiratory tract exposed to air pollution responded with more severe changes in men than in women [7-9]. The aim of this study was to compare cytological changes in sputa, depending on the sex of young adults exposed to a high level of urban air pollution, and to examine whether a big difference in duration of exposure has an effect on these relations. MATERIALS AND METHODS: The subjects in the study were young adults, university students (21 to 25 years of age), all non-smokers. At the time of sputa collection (March-April 1991) they were clinically healthy with no history of chronic and serious acute respiratory diseases. They were divided into two groups according to the duration of air pollution exposure. The first group included those who had been living in Sarajevo since birth. The second group included those who used to live in pollution-free areas, but for the last two to three years had been living in Sarajevo where they came to attend the University (Table 1). Specimens were early morning spontaneously produced sputa, expectorated directly into Saccomanno's fixative and after centrifugation immersed in paraffin. Nine sections from each block were cut and stained with haematoxylin and eosin. The differences between cytological findings were tested with Hi2 test and Fisher's exact test, regarding sex and duration of exposure to air pollution. RESULTS: The findings are summarized in Table 1. As many as 63% and 64% of the subjects from the first and the second group, respectively, were able to produce sputum, while the others produced saliva only. All sputa contained abnormal bronchial columnar cells and half of them contained respiratory spirals (Figures 1 and 2). Squamous metaplastic cells were found in 7% and 9% of subjects in the first and second group, respectively. These cells showed no evidence of atypia. Statistical tests failed to identify significant differences in the incidence of cytological findings between the groups regarding both the sex of the subjects and duration of their exposure to air pollution (Table 2). DISCUSSION: The ability to produce sputum with the frequent presence of respiratory spirals in both groups of our subjects indicate an extremely harmful effect of smog. However, the incidence of squamous metaplastic cell findings was less severe comparing to adults and the elderly urban population. In the elderly (65 to 105 years of age) severe epithelial changes developed spontaneously in the absence of any other known aetiological factor [8, 9]. The increase in severity of changes in the course of aging, in our opinion, due to a gradual decline in efficiency of the defence mechanisms and regenerative potential of the respiratory system. The experimental study showed a decline of the efficiency of mucociliary clearance [14] as well as the damage of the function and decrease in number of alveolar macrophages in lung parenchyma and bronchoalveolar lavage [15, 16]. Trying to answer the question why the sex related differences in the severity and incidence of epithelial changes appear in the groups of adults and the elderly, it is presumed that in aging the defence and regenerative potentials decline more rapidly in men than in women. (ABSTRACT TRUNCATED) PMID- 10377834 TI - [Aortic wall distensibility and the structure and function of the left ventricle in aged persons with isolated systolic hypertension]. AB - The number of old persons (over 65 years) with arterial hypertension is in a steady increase [1]. Such finding is mainly related to patients with isolated systolic hypertension. They present more than 60% of old persons with arterial hypertension [2]. Isolated systolic hypertension can be defined as increased systolic blood pressure to the value more than 160 mmHg and diastolic pressure to 90-95 mmHg [4, 5]. It has been suggested that the pathologic basis of this entity is in a decreased distensibility of aorta and great arteries. In patients with isolated systolic hypertension we studied the correlation between decreased aortic distensibility and systolic arterial blood pressure value. We also evaluated changes in the left ventricular structure and function during this type of hypertension. PATIENTS AND METHODS: We examined 59 patients older than 65 years. They were divided in two subgroups. First subgroup: 38 patients (74 +/- 11 years) with isolated systolic hypertension (ISH) and the second subgroup: 21 normotensive persons (NT) (73 +/- 6 years). Aortic distensibility was calculated by the formula: Aortic dystensibility = difference between aortic diameters/diameter aortae in diastole x pulse pressure. The ascending aortic diameters were measured 4 cm above the aortic valve by two dimensional echocardiography and pulse pressure was measured simultaneously by sphingomanometry. Using M-mod and two-dimensional echocardiography we measured end-diastolic (EDD) and end-systolic (ESD) left ventricular diameters and thickness of interventricular septum (IVS) and posterior wall (ZZ). We calculated the ejection fraction (EF) using Teichole formula. Changes in left ventricular structure were expressed by sum of interventricular septum and posterior wall thickness and left ventricular mass. We calculated left ventricular mass using the following formula: MLK = /EDD + IVS + ZZ)3 - EDD/ x 1.05. By pulse Doppler echocardiography we measured the peak velocity of the left ventricular filling (VE) and calculated the ratio between early and late peak velocity (VE/VA). RESULTS: Aortic distensibility was significantly lower in patients with isolated systolic hypertension than in normotensive subjects (0.10 +/- 0.02 x 1/100 1/mmHg vs 0.24 +/- 0.04 x 1/100 1/mmHg; p < 0.05). Such findings are presented in Table 1. At the same time, we found a significantly inversed correlation between aortic distensibility and systolic blood pressure value in patients with isolated systolic hypertension (r = 0.67; p < 0.05). From Table 2 it is visible that there were no significant differences between left ventricular diameter and mass in hypertensive patients. The sum of interventricular septum and posterior wall thickness was significantly greater in patients with isolated systolic hypertension than in normotensive patients (2.19 +/- 0.5 cm v.s. 1.93 +/- 0.4 cm; p < 0.05). This finding is also presented in Table 2. We found no statistically significant differences among the ejection fraction values in the studied subgroups (Table 3). The peak velocity of early filling and the ratio of early to late peak velocities were significantly lower in the hypertensive subgroup (0.4 m/s v.s. 0.54 m/s; p < 0.05; 0.69 v.s. 0.76; p < 0.05) (Table 3). DISCUSSION: In old persons with isolated systolic hypertension we found that aortic distensibility was significantly lower in comparison to normotensive subjects of the same age. Such finding supports the hypothesis that the reduced aortic distensibility is the cause of isolated systolic hypertension. At the same time, we found the inversed correlation between aortic distensibility and the mean systolic blood pressure value. Aging has an effect on reduction of aortic and great vessels distensibility. Thus, it causes arterial hypertension which changes the elastic properties of aorta. It is still questionable in which degree the systolic blood pressure value compromises the elastic properties of aorta. (ABSTRACT TRUNCATED) PMID- 10377837 TI - [The effect of the number of peritonitis episodes on peritoneal membrane function]. AB - INTRODUCTION: The problem of an adequate peritoneal dialysis has attracted attention in the recent years. The monitoring and adjustment of intraperitoneal dialysis liquid volume and dialysis duration between the filling and emptying, contributed to individualization of dialysis, improvement of its quality and prolongation of its application. Causes of decline of peritoneal dialyses adequacy have been recently the subject of many clinical studies. The aim of this study was to determine whether the number of peritonitis episodes and duration of treatment with continuous ambulatory peritoneal dialysis (CAPD) influence dysfunction of the peritoneal membrane. PATIENTS AND METHODS: We analysed 10 patients (4 females and 6 males), aged 27 to 77 years (60.1 +/- 13.9 years), who had been on CAPD for more than two years. All patients were tested at the beginning of CAPD, after one year and after two years of CAPD treatment (using 8 L exchanges per day): dialytic solute urea and creatinine clearance, daily and weekly; KT/V, daily and weekly; residual urea and creatinine clearance; and sum urea and creatinine clearance. The results are expressed as mean +/- SD and the differences between groups were studied by T-test and linear correlation coefficient. RESULTS: Daily and weekly KT/V declined during time, but was not statistically significant (Table 1). Decline of diuresis influenced the residual and sum creatinine and urea clearance, with no statistical significance (Table 1). Positive correlation was established between decline of diuresis and sum creatinine and urea clearance after one year of CAPD treatment (rcr = 0.7705; rur = 0.7782), as well as after two years (rcr = 0.6332; rur = 0.6852), with statistical significance (p < 0.05). During the first year of the study our patients had 20 episodes of peritonitis (2.00 +/- 1.65) and after two years this number increased to 51 (4.42 +/- 1.72). Negative correlation was evidenced between the number of peritonitis episodes and daily and weekly KT/V, with no statistical significance after one year (rd = -0.2462; rw = -0.2371), but with statistical significance after two years (rd = -0.6332; rw = -0.6852) of CAPD treatment (p < 0.05) (Graph 1). DISCUSSION: In order to achieve a better prognosis and longer CAPD therapy, we must take into account the patients age, nutrition, catabolism of proteins, residual renal function, and adequacy of dialysis. The consequences of an inadequate therapy can be reflected on morbidity and mortality in these patients. In spite of many discussions, the problem of an adequate peritoneal dialysis, and the value of daily and weekly KT/V are still controversial. Burkat recommends a daily KT/V above 0.24 as adequate and we found this KT/V value as appropriate in one half of our patients. The other authors found that adequate dialysis was achieved with KT/V above 0.29, what we found in two of our patients. The mean daily KT/V was 0.27 at the beginning of the study, and 0.26 after one year of treatment (Table 1); this finding is the the same as in the study of Gotch, and better than reported by Teehan et al. After two years of CAPD the mean daily KT/V was 0.23, what was better than in the study of Teehan et al. Many authors agree that weekly KT/V has to be at least 1.7 for an adequate dialysis; we found the same in one half of our patients in each time interval. Recently, many authors found that the optimal weekly KT/V was over 2, what we found in two of our patients. The mean weekly KT/V at the beginning of the study and after one year of CAPD was above the minimum of adequacy (Table 1). However, after two years of dialysis the mean weekly KT/V was 1.62; this was below the minimum of adequacy. At the beginning of dialysis the residual renal function declines, what we observed in our patients, but without statistical significance (Table 1). The decline of diuresis reflects on the residual and sum creatinine and urea clearance. (ABSTRACT TRUNCATED) PMID- 10377836 TI - [Hereditary deficiency of antithrombin III, protein C, protein S and factor XII in 121 patients with venous or arterial thrombosis]. AB - INTRODUCTION: Hereditary thrombophilia is caused by various inherited disorders which lead to familial tendency to recurrent venous thrombosis usually at an early age and with spontaneous onset. In the studies reported so far, the different prevalence of hereditary thrombophilia among patients with venous thrombosis was found, greatly depending on criteria for selection of patients. Arterial thrombosis is most often the consequence of arteriosclerosis but the prevalence of hereditary thrombophilia among young patients with arterial thrombosis and without recognized risk factors for arteriosclerosis is not known . In this study, the frequency of hereditary deficiencies of antithrombin III (AT III), protein C (PC), protein S (PS), plasminogen (PLMG), factor XII (F XII) and dysfibrinogenaemia was investigated over a 2-year period in 121 patients with venous or arterial thrombosis selected according to the recommendations of the British Committee for Standards in Haematology. PATIENTS AND METHODS: The study included total a of 121 patients (58 males and 63 females) with documented venous or arterial thrombosis. Table 1 shows patient's characteristics regarding gender, age and clinical manifestation of thrombosis. Each patient fulfilled at least one of the following criteria: a) venous thrombosis prior to the age of 45; b) arterial thrombosis prior to the age of 30, without risk factors for arteriosclerosis; c) recurrent thrombosis; d) familial tendency to thrombosis; e) thrombosis of unusual localization. A detailed history was taken from each patient on earlier personal or familial occurrence of thrombosis. For the purpose of this study, thrombophilia was characterized as congenital when the deficient protein was constantly below normal value and when the same deficiency was confirmed in a close family member; acquired when the acquired disorder predisposing to thrombosis was present in absence of constant protein deficiency; and idiopathic when the cause of thrombosis was unknown. All tests were performed in plasma obtained after centrifugation of venous blood anticoagulated with 0.129 mol/1 sodium citrate. Concentrations of fibrinogen, PT, PTT and F XII were measured by standard clotting methods. At III, PC and plasminogen activity were determined by chromogenic methods using commercial reagents (Boehring, Marburg, Germany). AT III, PC and total PS antigen were assayed by Laurell immunoelectrophoresis. The presence of lupus anticoagulant was investigated by recommended tests. RESULTS: A total of 15 patients (12.4%) fulfilled criteria for hereditary thrombophilia. Seven of them (5.8%) had AT III deficiency, five (4.1%) PC deficiency, two (1.6%) PS deficiency, and one patient had F XII deficiency. Secondary thrombophilia was found in 21.5% of patients and the cause of thrombosis in 66.1% of patients was not elucidated. A high frequency of hereditary thrombophilia has been found in patients with arterial thrombosis (40%). Among patients with hereditary thrombophilia thrombosis occurred at significantly younger age (29.9 vs. 42.2 and 40.9 yr.) compared to the patients with secondary and idiopathic thrombophilia, respectively. Patients with hereditary thrombophilia had also a higher occurrence of positive family history related to thrombosis (66.7% vs. 7.7% and 27.5%). DISCUSSION: The prevalence of hereditary thrombophilia in nonselected patients with venous thrombosis is relatively low, and for that reason the selection of patients, according recommended criteria, in whom the screening tests for congenital thrombophilia should be performed, is strongly suggested by many authors. In our study we used the generally accepted recommendations for investigation of patients with venous and arterial thrombosis. The presence of congenital thrombophilia was found in 15 (12.4%) of 121 studied patients, what is in accordance with results of other similarly designed studies. (ABSTRACT TRUNCATED) PMID- 10377838 TI - [Surgical treatment of secretory otitis media: persistent perforation as a rare complication]. AB - INTRODUCTION: The persistent perforation of the ear drum may appear as a complication after spontaneous or instrumental elimination of ventilatory tubes during surgical management of secretory otitis media. According to recent literature data its incidence can vary in an average interval from 0.5 to 3.4% for the short term ventilatory tubes and more than 8.5% for long term ventilatory tubes. This study showed the results of analysis of the incidence and therapy of persistent perforation of the ear drum after elimination of the short term ventilatory tubes during the treatment of secretory otitis media in 1554 children, treated over the period from 1988 to 1997 at the Ward of ORL, Zvezdara Clinical and Hospital Centre in Belgrade. MATERIAL AND METHODS: The retrospective analysis of medical documentation showed that 1554 children were surgically treated by short term ventilatory tubes between 1988 and 1997. Bilateral insertion of the ventilatory tubes was performed in 1353 children (Sheppard short term tubes) and unilateral insertion was performed in 201 children. The age of the patients ranged from 9 months to 12 years. RESULTS AND DISCUSSION: After 2907 inserted short term ventilatory tubes 25 (0.86%) persistent perforations of the ear drum were diagnosed. The average time of elimination was 6-8 months after tubes insertion. In 3 cases, refreshment of the edges of perforation and silastic sheet placement led to complete closure of ear drum defect. In 16 patients myringoplasty was performed. According to literature data and our clinical experience, the main cause of this complication is the state of the ear drum which is a part of pathophysiological mechanisms in secretory otitis media. The histopathological studies of the ear drum in secretory otitis media confirmed various morphological changes: thickening of the ear drum, subepithelialo oedema, infiltration of submucous space by lymphocytes, macrophages and plasmocytes. Fibrous layer showed irregularly oriented fibroblasts and a high production of fibrous fibres. The interaction among extracellular tissue, cells of the submucous and subepithelial layer and cytokines causes the process of reparation of the ear drum. CONCLUSION: Persistent perforation of the ear drum is a rare complication during surgical treatment of secretory otitis media, but it has its great clinical importance if spontaneous repair does not appear. Depending on the mucosal status, ear discharge, Eustachian tubes function and immunological status or upper respiratory mucosa, myringoplasty is the method of therapeutical choice. PMID- 10377839 TI - [Mastocytes in chronic inflammation of middle ear mucosa]. AB - INTRODUCTION: Pathohistological studies have shown that in secretory otitis media an increased number of mast cells have been found in mucosa of the middle ear and an increased level of histamine in the cavity of the middle ear. The aim of this study was to analyse the distribution and functional state of mast cells in middle ear mucosa biopsies in patients with chronic otitis media. MATERIAL AND METHODS: The pathohistological analysis was performed on 118 biopsy specimens of mucosa of the middle ear. The samples were taken intraoperatively from 39 patients during the surgical treatment of chronic otitis media with and without cholesteatoma and otomastoiditis. The specimens were taken from mastoid, attic space, promantory and protympanon. All biopsy specimens were prepared for hematoxylin-eozin staining and were identified with PAS method and Lennert Giemsa histamine granules. The Alcian blue-Safranin method was used to identify heparin granules in mast cells. By using the semiquantitative method, distribution of mast cells was marked as rare (+), numerous (+2) and high (+3). Also the number of degranulated cells was identified (Tables 1, 2). RESULTS AND DISCUSSION: The analyses of biopsy specimens of mucosa of the middle ear in chronic otitis media showed that cells were present in all areas of mucosa in a considerably higher number (Table 1). Degranulated cells were frequently found in biopsy specimens of otomastoiditis and contained predominately histamine granules; thus specimens taken from chronic otitis media, with and without cholesteatoma, showed mast cells with heparin granules with low rate degranulation. Pathophysiological examinations of the role of mast cells in the mechanism of chronic inflammation have shown that mast cells display through mediators the biphasic effect. They can participate in the process of deterioration of inflammation, whereby the released mediators cause the processes of tissue destruction, and can stimulate the process of tissue repair. Heparin accelerates the healing process of tympanic membrane perforation, stimulates the proliferation of endothelial cells and supports angiogenesis in the lamina propria of the tympanic membrane. Further studies are needed to clarify the role of histamine and heparin in the pathophysiology of chronic inflammation in chronic otitis media. PMID- 10377840 TI - [Conventional carotid endarterectomy]. PMID- 10377841 TI - [Cytokines in allergic inflammation]. PMID- 10377842 TI - [Drug therapy of hormone-dependent tumors]. PMID- 10377843 TI - [Thrombosis of the transverse sinus of the brain as a manifestation of antiphospholipid syndrome]. PMID- 10377844 TI - [Radiographic changes in patients with diabetes mellitus and pulmonary tuberculosis]. PMID- 10377845 TI - [Anniversaries of the Serbian Medical Society, 1962: 90 years since the founding of the Society and its realization and recognition]. AB - The 90th anniversary of the Serbian Medical Association was celebrated at the end of a period that surpassed the previous achievements of the Association--a system of medical protection was built, medical schools in Nish and Novi Sad were founded, and new medical institutions were erected. The construction of a system of medical protection was linked to the "social system, which in itself provides further development of the system in the direction of encompassing the total population". One could also detect in this intention an uncritical transfer of the concept of the National Health Service of Great Britain, when the requirements, health conditions and aging of the population were not similar to the present population. The Managing Board of the Serbian Medical Association of that time founded the Section of general practitioners and proposed the introduction of specialization of general medicine. The jubilee is preceded by years in which new medical institutions were built and medical schools founded in Nish and Novi Sad. The role of some members of the Serbian Medical Association and of the Managing Board was prominent in some documents dating only a couple of decades after these events. Certain documents and monographs published in celebration of an anniversary of the School of Medicine or of some medical institutions no longer mention the role of the Association. This is also the case with a monograph published in celebration of the 45th Jubilee of the Institute of Mother and Child Health Protection. That monograph does not include the activties of the Association and the first resolution of the Serbian Medical Association after the liberation, in 1946, was related to the problem of medical protection of the young "as the most important task of popular medical policy". In the monograph about the foundation of the Medical School in Novi Sad, the only connection with the past is the mention of date when the Medical School was founded. The 90th anniversary of the Serbian Medical Association, when some of the members were honored for their contributions, is the only anniversary in the long history of the Association. In total 47 doctors were honored. On the same occasion the Serbian Medical Association honored 92 doctors with memorial plaque for the "contribution to the activity of the Association". PMID- 10377846 TI - [Perfume allergy is not only a luxury problem]. PMID- 10377848 TI - [Forensic genetics]. PMID- 10377849 TI - [Perfume allergy]. PMID- 10377847 TI - [Elderly as a category. Age limits, discrimination, marginalization and polarization]. PMID- 10377850 TI - [Amyloidosis. A review]. AB - Amyloidosis is a heterogenous group of diseases, all characterized by extracellular deposition of amyloid either systemically or localized. Of wellknown diseases are Alzheimer's dementia, AL-amyloidosis (e.g. in multiple myeloma) and AA-amyloidosis (e.g. in rheumatoid arthritis). Amyloid is composed of three components of which the fibrillary component is the basis of amyloid classification. Many types of amyloid have a systemic distribution and give rise to varying symptoms. The diagnosis is based on biopsy, preferably of abdominal subcutis. The prognosis is poor, however, recent investigations on the three dimensional structure of the P-component provide hope for future therapy. PMID- 10377851 TI - [Scleroderma--systemic sclerosis. Serology, lung function and survival]. AB - Patients with systemic sclerosis (SSc) were studied with regard to the presence of antinuclear antibodies (ANA) and their clinical correlates (n = 230), pulmonary function (n = 176), and mortality and causes of death (n = 344). ANA were found in 85%. Anti-centromere antibodies were found in 34%, anti-Scl-70 antibodies in 13% and anti-U1-RNP antibodies in 6.5%. These serological groups were associated with limited SSc, diffuse SSc, and myositis/arthritis, respectively. The most prevalent finding at first lung function test was isolated reduction of diffusion capacity (47%). Further deterioration of diffusion capacity was related to the presence of anti-centromere antibodies and increased sedimentation rate. The standardized mortality rate (SMR) was 2.9, higher in young patients (SMR = 13) and patients with diffuse SSc (SMR = 4.5). PMID- 10377852 TI - [Follow-up of metatarsal fractures--treated with pressure bandage and weight bearing]. AB - We constructed a follow-up study of 61 patients, who had been treated for a simple metatarsal fracture in the A & E department two to three years earlier. All patients were treated with pressure bandage for three weeks and full weight bearing, no plaster was given. Forty-nine patients filled in a questionnaire and 37 patients, with 41 metatarsal fractures, were examined clinically and radiologically. The patients had pain for a median of 10 days and were walking with full weightbearing after a median of 21 days. Two percent (one patient) found the treatment not satisfactory. There were no clinical signs of malfunctioning of the feet. Of the fractures, 51% were avulsion fractures of the 5th metatarsal, 27% were fractures of the distal diaphysis of the 5th metatarsal, 10% were Jones' fractures and 12% other metatarsal fractures. All fractures healed without further treatment. We recommend pressure bandage and full weight bearing as first choice treatment of simple metatarsal fracture (2nd-5th metatarsal). PMID- 10377853 TI - [Neonatal asphyxia--prognosis based on clinical findings during delivery and the first day of life. A retrospective study of 54 newborn infants with asphyxia]. AB - A retrospective study of a cohort of 54 term infants with perinatal asphyxia admitted to the neonatal intensive care unit at Hvidovre University Hospital i 1995 is described. The purpose of the study was to find clinical markers for prediction of outcome after perinatal asphyxia. Neither complications of pregnancy, gestational age, the sex of the infant, passage of meconium before delivery, abnormal fetal heart rate nor birth weight seemed to have any interrelationship with outcome. The Apgar score was more depressed, and the metabolic acidosis worse among the infants with poor outcome. The best predictor of outcome after intrapartum asphyxia was the severity of postasphyxial encephalopathy. No infant with mild or moderate postasphyxial encephalopathy died or developed any handicap. All with severe postasphyxial encephalopathy either died or developed minor handicaps. PMID- 10377854 TI - [Suicide among patients with apoplexy. An epidemiological study]. AB - The purpose of this study was, on the basis of a cohort of patients with a discharge diagnosis of stroke (ICD8 code: 430-438) diagnosed in the period 1.4.1973-31.12.1989 in a representatively selected area of Denmark (County of Funen), to estimate whether or not the risk of suicide in stroke patients was increased compared to the background population, i.e. the total population of the County of Funen. The patients were followed for causes of death until end of 1989. Standard Mortality Ratios for suicide standardized for age and sex in male and female stroke patients were calculated. A total of 37,869 stroke patients were included in the study, 140 committed suicide in the study period (80 females and 60 males). Standard Mortality Ratio for suicide was significantly increased for all stroke patients. For women below 49 years and from 50-59 years Standard Mortality Ratios were 1376 and 1378 respectively. For men below 49 years and from 50-59 years Standard Mortality Ratios were 656 and 580 respectively. The suicides did not occur at any specific time-point after the stroke. The results of this study emphasize the need, concurrently with improvements in prevention and treatment of stroke, to improve the care of patients suffering from the impairment of a stroke. This is a continuous process, and studies are needed in order to decide how this is best done. PMID- 10377855 TI - [Amyloidosis: rapid course of AL-amyloidosis in a patient without myelomatosis]. AB - A case of severe AL-amyloidosis in a 72-year-old man is presented. Amyloid was especially deposited in the kidneys and the liver. The patient died one year after diagnosis with impaired kidney function and severe ascites. Despite the type of amyloid (AL-type), multiple myelomata or lymphoproliferative disease could not be demonstrated. PMID- 10377857 TI - [Psychopharmacological treatment of social phobia]. PMID- 10377856 TI - [Cerebral venous thrombosis]. AB - An 18-year-old woman with extensive cerebral venous thrombosis is described. The symptoms were fever, headache, nausea, vomiting and focal deficits. The diagnosis was confirmed by CT and MRI. Anticoagulation was given. Full restitution was achieved within three months. Possible predisposing factors were use of oral contraceptive, smoking and anticardiolipin antibodies. The causes, symptoms, investigations and possible modes of therapy are discussed. PMID- 10377858 TI - [Guidelines on registration of diagnoses and surgical interventions]. PMID- 10377859 TI - [Thrombolysis in acute apoplexy--more research is required]. PMID- 10377860 TI - [Apoplexy and thrombolysis]. PMID- 10377861 TI - [Neurotrophic factors and ALS]. PMID- 10377862 TI - [New priority trends in ocular traumatism]. AB - Types of ophthalmic traumas subject to change at present are enumerated: communal, criminal, war traumas, and injuries resultant from emergency situations (disasters). The changes are discussed. Main factors which should be borne in mind when investigating the problem of ocular injuries are determined. Priority trends of research in ophthalmic traumatology are defined. PMID- 10377863 TI - [Indications for transvitreal removal of foreign bodies from posterior eye segments]. AB - Results of treatment of 40 patients (43 eyes) with foreign bodies in the posterior segment of the eye are discussed. Special attention is paid to correct assessment of the risk and efficacy of removal of a fragment in each case. Only 67.5% of patients presented with obvious indications for removal of a foreign body through the vitreous. Fragments were removed in 83.3% of these patients. Complications occurred in 6.6%. The main causes of failure of sparing removal of a foreign body from the posterior segment of the eye were detachment of the retina and hemophthalmia. PMID- 10377864 TI - [Contusion changes in ophthalmic tonus: clinical observations and pathogenesis]. AB - Contusions of the eyeball involve reactive or secondary changes in intraocular pressure. Clinical examinations of 151 patients with contusion disorders in ophthalmic tone revealed reactive hypertension in 6.62%, reactive hypotone in 31.79%, secondary hypertension in 48.34%, and stable hypotone of the injured eye in 15.89% cases. Secondary changes in intraocular pressure were caused by massive intraocular hemorrhages and structural injuries to the eyeball. The main cause of reactive hypotone resulting from blunt injury to the eye was exfusion, detachment of the vascular membrane diagnosed at that period by ultrasonic examination in 76.92% cases. The main factors responsible for the postcontusion reactive syndrome are changes occurring within a very short period of time, intraocular pressure differences, and "loss" of the threshold volume of the anterior chamber humor because of its discharge during the shock, and activation of the kallikrein kinin system with release of kinins causing capillary dilatation and increasing capillary wall permeability in the presence of low activity of angiotensin converting enzyme which destroys bradykinin. PMID- 10377866 TI - [Role of ciliary muscle in ocular physiology and disease]. AB - The ciliary muscle is the largest intraocular muscle. It is characterized by a complex structure, rich blood supply, and high activity, particularly in the daytime, when visual work is performed, and even during night sleep. The activity of the ciliary muscle involves the adjacent structures. In fact, it continuously massages the anterior and median segments of the eye. Presbiopia leads to a decrease in the activity of the ciliary muscle, deteriorates blood circulation in it, and accelerates age-associated involution processes. Presbiopia is believed to be a risk factor for glaucoma, cataract, and some other dystrophic processes. The role of the ciliary muscle in health and disease is still to be investigated. PMID- 10377865 TI - [Significance of biochemical values of lacrimal fluid for early diagnosis and prognosis of traumatic uveitis]. AB - Parameters reflecting the severity of hypoxia and membrane destruction in the lacrimal fluid correlate with the severity of injury to uveal tract tissues. PMID- 10377867 TI - [Regularities of corneal tissue correlation and their biological significance for regeneration and pre-tumor growth]. AB - Experimental denervation of the central portions of the cornea in rabbits permitted us to hypothesize "autonomism" of corneal tissues, manifesting as soon as corneal tissues loose connection to the nervous system inhibiting cell growth and division. It involved edema, decreased differentiation, loss of correlation between the epithelium and stroma, submerged growth, and eventually even transformation into a sort of tissue culture in the organism. Experiments demonstrated that increased tissue differentiation is paralleled by in-growth of nerve endings. The degree of differentiation and functional intactness of tissues is maintained by the nervous system. If there are no conditions for reinnervation or the injury (irritation) is repeated, the process transforms into pathological: cords of immature tissue penetrate into the depth of the eye from open edges of corneal and scleral wounds, granulation of chronic wounds is abnormal, and pretumor states of tissues develop. Further status of such tissue apparently depends on new vessels, repeating the transformation of epithelial stroma and connective tissue. Increased growth of the epithelium and stromal elements leads to thickening of vascular wall, dilatation of vascular lumen, and to complete obliteration. Repeated irritation resumes the process of tissue transformation with formation of new vessels. Angiogenesis is apparently related to denervation, and each new irritation induces repeated vascular growth. Treatment and prevention are to be aimed at removal of the irritants and improvement of the nervous system function maintaining the maturity of tissues in an organism. PMID- 10377868 TI - [In situ laser keratomileusis for correcting myopia and astigmatism of different severity]. AB - Myopia and astigmatism were corrected by laser specialized keratomylesis in 1357 eyes with myopia of 1-18.0 diopters and astigmatism up to -5.5 diopters. The proposed method is highly effective in myopia of different severity and astigmatism and is preferable to photorefraction keratectomy. PMID- 10377869 TI - [Clinical and immunological criteria for predicting the course of uveitis]. AB - Clinical and immunological parameters are studied in 205 patients with uveites of different etiology in order to define the criteria permitting the prediction of uveitis course and relapses. The prognosis is unfavorable for autoimmune conditions, including peripheral, systemic, syndromal, tuberculous uveites and mixed infections. Clinical cure is unstable in diffuse inflammatory changes, persistent cellular infiltration of the vitreous body, formation of cyclic membranes in it, in cataract complications, and exudative hemorrhagic chorioretinitis. High titers of antibodies to tissue antigens and deficiency or hyperimmunoglobulinemia A, discoordination of T and B immunity, and sensitization to several antigens are risk factors for relapses. Clinical and immunological parameters should be analyzed for predicting the disease course and prescribing the basic and antirelapse therapy. PMID- 10377870 TI - [Clinical and immunological factors in predicting an early exudative reaction after extraction of senile cataracts with implantation of elastic IOL]. AB - Clinical and immunological studies in 170 patients with senile cataracts carried out before and during the first 7 days after cataract extraction with implantation of intraocular lenses helped define the risk factors for development of an early exudative reaction: patients' age under 60 years, clinical signs of secondary immune deficiency presenting as infection syndrome, increased levels of nonspecific immunity factors with deterioration of the phagocytic component, T lymphopenia, CD4/CD8 imbalance, and increased levels of interleukin-1 beta and tumor necrosis factor in the serum and lacrimal fluid. PMID- 10377871 TI - [Energy metabolism disorders in experimental atherosclerotic chorioretinopathy]. AB - Electron-microscopic, electron-histochemical, and biochemical studies in 12 rabbits with hypercholesterolemia revealed disorders in energy metabolism and redox processes in endotheliocytes of choroidal microvessels. These disorders resulted in functional defects of endotheliocytes, with changes in their membrane structure and disorders in ocular membrane microcirculation. PMID- 10377872 TI - [Clinical assessment of infrasonic phonophoresis efficacy in the treatment of bacterial keratitis]. AB - Therapeutic efficacy of infrasonic phonophoresis is studied in 30 patients with bacterial keratitis. Control group consisted of 87 patients with the same diagnosis. Clinical studies included comparative evaluation of the therapeutic efficacy of infrasonic phonophoresis and traditional local instillations of the same drugs. Before treatment, visual acuity was the same in both groups, while after regression of inflammation after treatment it was 0.13 higher in the phonophoresis group. Results of clinical studies indicate a higher efficacy of infrasonic therapy of patients with keratitis. The duration of therapy was decreased, number of bed-days decreased, and visual acuity after treatment improved. PMID- 10377873 TI - [Locferon: new eyedrops for treatment eye adenoviral diseases]. AB - Locferon, interferon eyedrops, was used in the treatment of adenovirus diseases of the eyes in 68 patients. The results demonstrated the advantages of locferon over interferon. The drug was highly effective and duration of treatment decreased; it was well tolerated and caused no side effects or toxic allergic reactions. PMID- 10377874 TI - [Histochrome, a new antioxidant, in the treatment of ocular diseases]. AB - The efficacy of a new bioantioxidant histochrome is studied in 92 patients (98 eyes). The drug was injected subconjunctivally or parabulbarly in a dose of 0.5 ml 0.02% solution daily, 7-10 injections per course. The clinical effect of histochrome is determined by the localization, duration, and extent of the process. The drug was the most effective in the treatment of intraocular hemorrhages which occurred no more than 7 days before. Electroretinography demonstrated pronounced retinoprotective properties of the drug which improved the electrophysiological parameters in degenerative processes in the retina and optic nerve. Hence, histochrome is characterized by hemoresorption, retinoprotective, and antioxidant properties and is recommended for the treatment of ocular diseases involving metabolic disorders in the retina, vascular membrane, and cornea. PMID- 10377875 TI - [Retinal phlebothrombosis: modern aspects in etiology, pathogenesis, diagnosis and therapy]. PMID- 10377876 TI - [Contribution of immunology to the development of biomedical disciplines]. AB - The paper deals with the history of immunology, the contribution of Nobel prize winners to its progress. It analyzes the achievements of immunology and its contribution to the development of basic biology, biotechnology, medicine, and agriculture. The greatest attention is paid to latest developments which are of high scientific, social, and economic values. These include the interpretation of the structure of immunoglobulins, the mechanisms responsible for the development of immunological responses, as well as hybridome immunobiotechnology, advances in immunogenetics, immunopharmacology, clinical immunology. The Russian developments of priority are the design of new-generation immunotoxins, the clinical use of the new-generation immunomodulators having a controlled structure, such as polyoxidonium, an nontoxic biodegraded synthetic polymer that has an immunostimulating action and detoxifying activity, the discovery of bone marrow bioregulatory molecules, the design of myelopid, their based immunomodulator showing its analgetic opiate-like action, the isolation, purification, synthesis, and characterization of individual myelopeptides (MP-1 to MP-6) which are constituents of myelopid. PMID- 10377877 TI - [Genetics of immune response and vaccines of tomorrow]. AB - The paper deals with the problems in the designing and development of fundamentally new conjugated antigen (hapten, peptide, protein)-polymer immunogens and vaccines and with their introduction into clinical practice. The history of designing conjugated and synthetic immunogens is briefly given. A new principle in the design of artificial vaccinating preparations is considered. The results of study of a genetic control of an immune response to artificial antigens are analyzed. The properties of high-molecular polyions, the molecular and cellular mechanisms of phenotypic correction of genetic control of an immune response to conjugated antigen-polymer complexes are shown. The artificial antigens and vaccinating preparations based on artificial polyions and designed by the author and his colleagues are described. Particular attention is paid to new designed vaccinating preparations, including Grippol which has been permitted for clinical application and manufacture. PMID- 10377878 TI - [Neuroimmunopathology]. AB - The paper deals with a part of neuroimmunopathology which is concerned with the role of the pathologically altered nervous system in the pathology of the immune system and vice versa and considers disturbances in the interconnection of nervous and immune systems. Antibodies against neuromediators and nervous tissue can produce both pathogenetic and sanogenic effects depending on the neurotransmitter function or the nervous structure which they are specifically directed towards. The effects of systematically administered antibodies against nervous tissue and neuromediators demonstrate that antibodies can penetrate into the central nervous system (CNS). The adoptive transfer of the signs of a pathological condition by immunocytes from CNS abnormalities illustrates the role of the immune system in nervous diseases. PMID- 10377879 TI - [Immunology of cancer]. AB - The paper present a brief review of progress in cancer immunology in the past 50 years and its use in the immunodiagnosis, immunoprevention, and immunotherapy of cancer. Tumor markers have been a basis of the serological diagnosis of some tumors and the immunophenotyping of leukemia. Hepatitis B virus vaccination should reduce the incidence of hepatic cell carcinoma. Investigations have provided impetus to the design of molecular genetic anticancer vaccines which are being tested in the clinical setting. PMID- 10377880 TI - [Cell interaction in immune response]. AB - The recognition of antigens by specific T- and B-lymphocytic receptors underlies an immune response. However, the formation of a potential signal for the activation of lymphocytes requires an additional their stimulation (costimulation). The main source of costimulation signals is the interaction of the surface molecules of lymphocytes and accessory cells. The interaction between the T-cell surface molecules CD28 and costimulatory molecules of antigen presenting cells (CD80 or CD86) is the most important point of the T-helper cell activation. The interaction between B-cell molecule CD40 and T-helper surface molecule CD154 is the key event of B-cell (and other antigen-presenting cell) activation. When costimulation is absent, antigen recognition induces specific lymphocytic anergy or apoptosis. Defects of costimulatory molecular expression or function can cause immunodeficiency. For example, hereditary defect of CD154 expression causes the hyper-IgM syndrome. The soluble forms of some costimulatory molecules are considered to be potential immunomodulators. PMID- 10377881 TI - [Natural cytotoxicity in complex of cell interactions]. AB - The paper summarizes the results of studies on the mechanisms and factors that do prevent natural killer (NK) cells from their cytotoxicity against the body's own cells. The findings support the fact that three-level mechanisms are involved in the protection of the body's cell from NK cell damage. The mechanisms of the first endogenic level are ensured by the factors which form a regulatory balance in the complex of NK cell responses. Those of the second endogenic level are ensured by the factors that participate in the transfer and exchange of metallic cations in the cell microenvironment. The third (exogenous) level mechanisms have been found in the study of the lymphocytic functions of cyanobacterial exometabolites and their derivatives from tap water probes. The products tested are able to block the generation of the free oxygen radicals utilized by NK cells in their lethal shock. A limitation of NK cell cytotoxicity seems to an important condition for non-cytotoxic regulatory NK cell interactions with immune and other cells. PMID- 10377882 TI - [Immunogenetic achievements for medicine]. AB - The paper presents the results of recent studies of human major histocompatibility complex. They are due to the progress made at the serological and molecular genetic levels of research and are applied in practical medicine, including in endocrinological and transplantological care. The main prospects of using these achievements in immunogenetics in the treatment of cancer and infectious diseases, including AIDS are defined. PMID- 10377883 TI - [Chronobiology of immune system]. AB - The biological rhythmological programme of the immune system is a constituent of the body's common biological rhythmological programme. Its pattern seems to be genetically determined and reflects the functional status of the system. The chronobiological mechanisms responsible for the regulation of immune functions lie in the presence of certain phasic interrelations between the biological rhythms of the synthesis and production of regulatory agents on the one hand, and those of the receptor system and metabolic potential of immunocompetent cells on the other. The facts given in the paper may be a basis for a chronobiological approach to better understanding the mechanisms of the physiology and pathology of the immune system. The medical significance of study of the structural and temporal pattern of the immune system consists in the development of new techniques for diagnosis, prognosis, therapy, and assessment of risk factors in immunopathological conditions. PMID- 10377884 TI - [Immunology of reproduction]. AB - The paper presents the results of studies of the role of immune factors in the pathogenesis of infertility, impaired gestation, and perinatal abnormalities, which have been performed at the Research Center of Obstetrics, Gynecology, and Perinatology, Russian Academy of Medical Sciences, in the past decade. It gives an evaluation of the contribution of antispermatic antibodies to infertility of unclear genesis, the systemic and local factors of immunity in females with endometriosis, the immune and interferon status in those with sexually transmitted infections, the role of immune factors and intrauterine infection in gestational abnormalities, the involvement of autoimmune mechanisms in reproductive disturbance. The trends of further studies are outlined. PMID- 10377885 TI - [Myelopeptides as a new group of regulatory peptides]. AB - The paper summarizes the data available on the structural and functional characteristics of the brain marrow immunoregulatory peptides myelopeptides (MPs). It describes the amino acid sequences of 6 individual MPs, their biological properties and mechanism. MP-1 is a hexapeptide having an immunocorrective effect. MP-2 has antitumor action, MP-3 stimulates the phagocytic activity of macrophages, M-4 is a new differentiating factor. The endogenous origin of the MPs isolated, their structure, and the pattern of their biological effects lead to the conclusion that MPs represent a new group of regulatory peptides that are promising for clinical application. PMID- 10377886 TI - [Interferon inducers: new generation of immunomodulators]. AB - Interferon (IFN) inducers are the agents having a wide range of antiviral activity. They have not only etiotropic, but marked immunomodulating effects. INF inducers different in its nature cause IFN synthesis in different immunocytic populations. The kinetics of the production of IFN and their antigenic composition depend on the chemical structure of an inducer and on their stimulated target cell populations. Their capacity of stimulating the synthesis of IFN in these or those organs determines the spectrum of application of an inducer and the time course of serum IFN accumulation--the strategy of its use. IFN inducers have a number of advantages over exogenous IFN. They stimulate the synthesis of the body's own IFN, which has no antigenicity unlike the most widely used recombinant IFN. IFN inducers cause IFN circulation at the therapeutical levels, at the same time to achieve such an effect requires multiple administration of large-dose exogenous agents. Some IFN inducers includes IFN synthesis in particular cell populations and organs, which has certain advantages over the polyclonal stimulation of IFN immunocytes. IFN inducers are well combined with IFN and other antiviral agents. The combination of these properties leads to the conclusion that IFN inducers are a highly promising group of agents having both etiotropic and immunomodulating effects. PMID- 10377887 TI - [Imunofan: new-generation synthetic peptide agent]. AB - The paper reviews the development, mode of action and field of application of synthetic regulatory peptides in the pathogenetic and immunocorrective therapy of infectious and noninfectious diseases. Great progress has been made in designing new-generation small regulatory peptides by modifying the sequence of amino acid residues in the active fragments of natural hormones to change their biological activity and therapeutic properties. The original hexapeptide Arg-alpha-Asp-Lys Val-Tyr-Arg has been designed by chemically modifying the thymic hormone Thymopoietin in positions 32-37. The agent has been called Immunofan. It is manufactured in ampoules containing 1 ml of 0.005% sterile solution for subcutaneous and intramuscular injections. The trials of Immunofan have demonstrated that it is able to restore cell immunity, the oxygen-dependent neutrophilic bactericidal system and antiviral antibody production. It decreases the levels of inflammatory mediators, such as TNF and IL-6, and activates the redox system. Included into the complex therapy of patients with cancer diseases, Immunofan enhances the body's reserve capacity to inactivate free radicals and oxidants, substantially shortens radiation and toxic reactions. Its use ensures the continuum of chemoradiotherapy. Used in the complex therapy for chronic infections (brucellosis, hepatitis B and C, opportunistic infections), Immunofan enhances antiviral and antibacterial immunity, shortens the manifestation of clinical symptoms and major syndromes of diseases. PMID- 10377888 TI - Target genes of homeodomain proteins. AB - Homeodomain proteins are transcription factors that share a related DNA binding domain, the homeodomain. This class of proteins was first recognized in the fruitfly Drosophila melanogaster where they cause homeotic transformations such as a fly with four wings instead of two (Lewis EB. A gene complex controlling segmentation in Drosophila. Nature 1978;276:565-570 [Ref. 18]). They are now known to exist in all eukaryotes where they perform important functions during development. Given that homeodomain proteins are transcription factors, they control the expression of downstream genes to regulate development. Which genes are controlled by homeodomain proteins and how many of them are there? This review focuses on a recent paper by Liang and Biggin (Liang Z, Biggin MD. Eve and Ftz regulate a wide array of genes in blastoderm embryos: the selector homeoproteins directly or indirectly regulate most genes in Drosophila. Development 1998; 125:4471-4482 [Ref. 1]), which proposes that the Drosophila homeodomain proteins Even-skipped and Fushi-tarazu directly control the expression of the majority of genes in the Drosophila genome. An alternative view, that most genes are only indirectly affected by homeodomain proteins is also discussed. PMID- 10377889 TI - Scrambled or bisected mouse eggs and the basis of patterning in mammals. AB - Several findings challenge the notion that specification of cell types and embryonic axes in mammals are rooted entirely in the temporal and spatial relations between cleaving blastomeres. They raise the question as to whether, as in most non-mammalian species, these processes depend on information already present in the egg. However, experiments designed to investigate this possibility directly by perturbing the organization of the zygote or, very recently, by deleting one or other of its polar regions [M. Zernicka-Goetz. Fertile offspring derived from mammalian eggs lacking either animal or vegetal poles. Development 1998;125:4803-4808 (Ref. 1)], have been interpreted to mean that such a role for the egg can be discounted. This conclusion seems premature in view of continuing uncertainty regarding the developmental potential of individual blastomeres in mammals. PMID- 10377890 TI - Genetics of epithelial polarity and pattern in the Drosophila retina. AB - This review is focused on recent advances in our understanding of the development of coordinated cell polarity, through experiments on the Drosophila compound eye. Each eye facet (or "ommatidium") contains a set of eight photoreceptor cells, placed so that their rhabdomeres form an asymmetric trapezoid. The array of ommatidia is organized so that these trapezoids are aligned in two mirror-image fields, dorsal and ventral to the eye midline (or "equator"). The development of this pattern depends on two systems of positional information that inform the cluster of cells that will form an ommatidium of anterior/posterior (a/p) and dorsal/ventral (d/v) direction. The former (a/p) is encoded by a progressive wave of development (the morphogenetic furrow). The latter (d/v) involves molecules known to act in tissue polarity in other organs and organisms. Our understanding of the function of these molecules rests not only on their mutant phenotypes, biochemistry, and expression patterns, but also on the spatial effects when mutant patches of cells are made (genetic mosaics). PMID- 10377891 TI - Defending genome integrity during DNA replication: a proposed role for RecQ family helicases. AB - The RecQ family of DNA helicases have been shown to be important for the maintenance of genomic integrity in all organisms analysed to date. In human cells, representatives of this family include the proteins defective in the cancer predisposition disorder Bloom's syndrome and the premature ageing condition, Werner's syndrome. Several pieces of evidence suggest that RecQ family helicases form associations with one or more of the cellular topoisomerases, and together these heteromeric complexes manipulate DNA structure to effect efficient DNA replication, genetic recombination, or both. Here, we propose that RecQ helicases are required for ensuring that structural abnormalities arising during replication, such as at sites where replication forks encounter DNA lesions, are corrected with high fidelity. In mutants defective in these proteins, not only is replication abnormal, but cells display aberrant responses to DNA-damaging agents or inhibitors of DNA synthesis. We suggest that RecQ helicases may be important for the integration of cellular responses to these insults, such as by linking cell cycle checkpoint responses to recombinational repair. PMID- 10377892 TI - New insights into the t-complex and control of sperm function. AB - The mouse t-complex, located on chromosome 17, contains genes known to influence male, but not female, fertility. Although some t-complex genes are recessive lethals, t-chromosomes are maintained in the population by transmission ratio distortion. When male mice heterozygous for the t-chromosome mate with wild-type females, most offspring will possess the t-chromosome, indicating a link between t-complex genes and sperm function. Several proteins coded for by t-complex genes have been localised in the sperm flagellum, suggesting roles relating to motility. Another t-complex protein appears able to regulate the adenylyl cyclase/cAMP signal transduction pathway, known to play an important role in capacitation. Defective motility and/or failure to capacitate ("switch on") would result in poorly fertile or infertile spermatozoa. Given the existence of human homologues for many genes in the t-complex and the prevalence of "male factor" infertility, information obtained about the t-complex not only will provide insight into basic biological mechanisms but may be of future clinical relevance as well. PMID- 10377893 TI - Deconstructing cell determination: proneural genes and neuronal identity. AB - Vertebrates express scores of bHLH proteins during neural development. Earlier studies inspired by the established role of "proneural" genes in fly neurogenesis, as well as by the vertebrate bHLH myogenic program, focused on the reconstruction of bHLH gene cascades, which are thought to control successive steps leading to neuronal differentiation. Little attention has been paid thus far to the relationship between the diversity of neural bHLH genes and the diversity of neuronal phenotypes. This article reviews recent evidence that, akin to their fly counterparts, vertebrate neural bHLH genes probably confer not only "generic" neuronal properties, but also neuronal type-specific properties, inextricably linking neural determination and the specification of neuronal identity. We also speculate on the relations between positional information and gene activity, and on the evolutionary significance of the diversity of bHLH genes. PMID- 10377894 TI - Activities of cold-shock domain proteins in translation control. AB - For efficient processing, transport, storage, translation, and degradation, stretches of RNA transcripts are required in a single-stranded conformation (ssRNA). A superfamily of OB-fold proteins is characterized by preference of binding to ssRNA. This superfamily consists of proteins containing either an S1 domain (S1-D) or a cold-shock domain (CSD). In a variety of situations. S1-D or CSD proteins are found in association with DEAD-box RNA helicases and the two types of protein appear to function together to maintain regions of ssRNA. CSD proteins are commonly found bound to stored (nontranslating) mRNA, particularly during early development. Although complete removal of the CSD proteins from mRNA permits its translation in vitro, low concentrations of CSD protein on the mRNA may be required for maximal translation efficiency in vivo. Another component of stored mRNP particles in Xenopus oocytes is the protein kinase CK2, which phosphorylates the associated CSD proteins. It is argued here that the loading of CSD proteins on mRNA and the stability of the protein/mRNA complex are regulated by RNA helicase activity and protein phosphorylation. PMID- 10377895 TI - Hierarchical differentiation of multipotent progenitor cells. AB - Selection of a pathway of differentiation in multipotent progenitor cells may depend on the amount of histone H1 or H1 zero relative to the core histones. With low levels of these linker histones, it is proposed that an evolutionarily more ancient cell differentiation occurs. Greater repetition of AT-rich regulatory motifs allows more frequent, hence earlier, transcription of genes, accounting for this type of cell differentiation. It is further proposed that a decrease of total cell protein accumulation leads to an increase of histone H1 or H1 zero relative to the core histones. It is suggested that these linker histones preferentially bind the more AT-rich regulatory sequences, thereby restricting the phylogenetically more ancient differentiation potency. This allows differentiation of an evolutionarily younger cell type. PMID- 10377896 TI - Instrumental biosensors: new perspectives for the analysis of biomolecular interactions. AB - The use of instrumental biosensors in basic research to measure biomolecular interactions in real time is increasing exponentially. Applications include protein-protein, protein-peptide, DNA-protein, DNA-DNA, and lipid-protein interactions. Such techniques have been applied to, for example, antibody antigen, receptor-ligand, signal transduction, and nuclear receptor studies. This review outlines the principles of two of the most commonly used instruments and highlights specific operating parameters that will assist in optimising experimental design, data generation, and analysis. PMID- 10377897 TI - The enigmatic megakaryocyte gradually reveals its secrets. AB - The recent cloning of thrombopoietin has brought many insights into the cellular and molecular mechanisms of megakaryocyte and platelet development. Thrombopoietin was cloned based on its binding to the product of the proto oncogene c-mpl and was found to affect all aspects of thrombopoiesis. Many of the molecular pathways that mediate thrombopoietin action have been discerned. Upon hormone binding, the megakaryocyte thrombopoietin receptor homodimerizes, activating members of the JAK family of kinases, which, in turn, phosphorylate the receptor, generating docking sites for second messengers that affect multiple signalling pathways. Ultimately, cellular proliferative and anti-apoptotic mechanisms are initiated, increasing megakaryocyte numbers, as are processes that uncouple DNA synthesis from nuclear and cytoplasmic division, generating polyploid cells. As the net result of thrombopoietin action is an expansion of cells that give rise to mature platelets, the availability of the recombinant hormone has provided new opportunities to manipulate blood cell development for therapeutic benefit. PMID- 10377899 TI - Surgery to prevent breast cancer? PMID- 10377898 TI - Genetics, eugenics and the medicalization of social behavior: lessons from the past. PMID- 10377900 TI - When heart patients need non-heart operations. PMID- 10377901 TI - Mind-body solutions for chronic pain. PMID- 10377902 TI - Nothing to lose but excess pounds. PMID- 10377903 TI - A brief history of depression. PMID- 10377905 TI - I have terrible low-back pain. Should I try magnets? PMID- 10377904 TI - Can nicotine help Parkinson's? PMID- 10377906 TI - What can be done for interstitial cystitis when oral medications and bladder instillations have not worked? PMID- 10377907 TI - Taking anger to heart. PMID- 10377908 TI - A "natural" remedy for high cholesterol. PMID- 10377909 TI - New solutions for persistent nerve pain. PMID- 10377910 TI - Are your vaccinations up to date? PMID- 10377911 TI - A brief history of antibiotics. PMID- 10377912 TI - I have atrial fibrillation and cannot take beta-blockers. What are my alternatives? PMID- 10377913 TI - Will you tell me more about the implants being used at Hopkins to restore voice in stroke patients? PMID- 10377914 TI - Short-sighted approach to long-term care. PMID- 10377915 TI - GI emergencies: rapid therapeutic responses for older patients. AB - Comorbid conditions and decreased physiologic reserves make persons age 65 and older particularly vulnerable to the adverse consequences of acute blood loss. Thus, gastrointestinal bleeding in this population presents an urgent clinical challenge. Treatment of such emergencies may be the preview of the gastroenterologist, but the primary care physician can play a key role in the preliminary diagnosis and management, timely referral for emergency intervention, and follow-up care. Because of age-related changes, including decreased cardiovascular reserve and a higher risk of hemorrhage, any evidence of GI bleeding in an older patient demands diligent intervention. PMID- 10377916 TI - Arthritis: new agents herald more effective symptom management. AB - For physicians and patients alike, managing the symptoms of rheumatoid and osteoarthritis is an ongoing challenge. Myriad therapies are available, although virtually all provide only temporary relief and produce side effects that interrupt long-term use. New disease-modifying antirheumatic drugs, biologic response modifiers, and cyclooxygenase inhibitors offer the promise of more effective, longer lasting symptom management and, in some cases, reduced side effects. The population of older persons affected by arthritis continues to grow. Increased familiarity with these new treatments will aid primary care physicians in helping older patients better manage their arthritis during the next decade. PMID- 10377917 TI - Diversity leadership. PMID- 10377918 TI - A shared statement of ethical principles for those who shape and give healthcare. A working draft from The Tavistock Group. PMID- 10377919 TI - Data collection for nursing work force strategic planning in California. PMID- 10377920 TI - Evaluating a medical day center in Hong Kong. Evidence-based management practice. PMID- 10377921 TI - Evidence-based clinical practice. AB - Healthcare professionals are trying to facilitate the use of evidence-based decision making for individual patients and patient populations they are privileged to serve. The authors describe an evidence-based multidisciplinary clinical practice model developed at the University of Colorado Hospital along with a clinical example of how the model was used to improve quality and decrease costs. PMID- 10377922 TI - Interpreter services in healthcare. Policy recommendations for healthcare agencies. AB - The author discusses the need for interpreter services for delivering effective healthcare in a country such as Canada, which is made up of people from diverse ethnocultural and linguistic backgrounds. As well as examining the complexity of allocation decisions and policy implementation, the author proposes policy recommendations for healthcare agencies to set up an integrated system of interpreter services based on cost-effective and equitable use of finite resources and partnership between agencies. PMID- 10377923 TI - Achieving information systems support for clinical integration. PMID- 10377924 TI - Epidemiologic determination of community-based nursing case management for stroke. AB - Efforts to control costs, especially those resulting from the 1997 Balanced Budget Act, have resulted in profound opportunities for futuristic community based nursing care. A retrospective chart review was conducted on 1,992 stroke patients discharged from 15 Cincinnati hospitals from July 1, 1995, to June 30, 1996. Determinants and descriptors of stroke distribution were identified. This study shows how nurses can plan cost-effective care while maintaining quality through an epidemiologic assessment of patient, family, and community needs. PMID- 10377925 TI - Implementation of the ANA report card. AB - A major challenge in healthcare today is measuring the quality of care. To explore nursing's contribution to patients in acute care settings, the American Nurses Association commissioned the development of the "Nursing Report Card." This study explored whether these report card indicators capture quality care. The convenience sample comprised 1,500 patients and 300 nurses from 16 units at an academic medical center. Using regression analysis, the most consistent predictor of outcome indicators was the percentage of RNs of the total staff. PMID- 10377926 TI - Informatics and process improvement. The clinical design unit. AB - Seeking to integrate departments and services, reduce costs, and enhance quality, administrators at a university medical center implemented a clinical design unit. The authors describe the operations of the unit in testing new processes before rollout to the entire center. PMID- 10377927 TI - Gastric stromal sarcoma, pulmonary chondroma, and extra-adrenal paraganglioma (Carney Triad): natural history, adrenocortical component, and possible familial occurrence. AB - OBJECTIVE: To investigate the natural history of the triad of gastric stromal sarcoma, pulmonary chondroma, and extra-adrenal paraganglioma, a rare syndrome of unknown cause primarily affecting young women. METHODS: Mayo Clinic records, the world literature, and the author's files were searched for patients with all or 2 of the 3 tumors. RESULTS: Seventy-nine patients, 67 women and 12 men, were identified, 17 (22%) with the 3 tumors and 62 (78%) with 2 tumors. Forty-two (53%) had gastric and pulmonary tumors, the most common combination. The longest interval between detection of the first and second components was 26 years (mean, 8.4 years; median, 6 years). Follow-up ranged from 1 year to 49 years (mean, 20.6 years; median, 20 years). Sixty-four patients (81%) were alive, 19 (24%) apparently free of disease and 45 (57%) with residual or metastatic tumors. Thirty-two patients (41%) had had 1 or more local recurrences of the gastric sarcoma; the longest interval to first recurrence was 36 years. Twenty-one survivors (27%) had hepatic metastatic gastric sarcoma with follow-up of 1 year to 25 years (mean, 9.3 years; median, 7 years). Fifteen patients (19%) were dead, 10 (13%) of whom died of the disorder. Ten patients (13%) had nonfunctioning adrenocortical tumors. Two patients each had a sibling with 1 component of the triad. CONCLUSIONS: The triad is a chronic, persistent, and indolent disease. Benign adrenocortical tumors are a component of the condition. The disorder may be familial. PMID- 10377928 TI - Staphylococcus aureus prosthetic joint infection treated with prosthesis removal and delayed reimplantation arthroplasty. AB - OBJECTIVE: To estimate in patients with Staphylococcus aureus prosthetic joint infection after total hip arthroplasty (THA) or total knee arthroplasty (TKA) the microorganism-specific cumulative probability of treatment failure after prosthesis removal and delayed reimplantation arthroplasty. PATIENTS AND METHODS: All patients with S aureus THA or TKA infection, according to a strict case definition, who were treated with prosthesis removal and delayed reimplantation arthroplasty at Mayo Clinic Rochester between 1980 and 1991 were identified. The study group comprised patients who were free of infection at the time of reimplantation arthroplasty. This cohort was followed up until treatment failure, infection with another organism, prosthesis removal, death, or loss to follow-up occurred. The Kaplan-Meier survival method was used to estimate the cumulative probability of treatment failure. RESULTS: Among 120 S aureus prosthetic joint infections treated with prosthesis removal during the study period, 38 episodes (22 THA, 16 TKA) in 36 patients met the study inclusion criteria. After a median of 7.4 years (range, 0.9 year-16.4 years) of follow-up, treatment failure occurred in 1 (2.6%) of 38 episodes 1.4 years after reimplantation arthroplasty. The 5-year cumulative probability of treatment failure was 2.8% (95% confidence interval, 0%-8.2%). CONCLUSIONS: These data suggest that prosthesis removal and delayed reimplantation arthroplasty is an effective treatment to limit the recurrence of S aureus prosthetic joint infection, provided there is no evidence of infection at the time of reimplantation arthroplasty. PMID- 10377929 TI - Should all Pima Indians with type 2 diabetes mellitus be prescribed routine angiotensin-converting enzyme inhibition therapy to prevent renal failure? AB - OBJECTIVE: To determine how effective angiotensin-converting enzyme (ACE) inhibitors must be in preventing diabetic nephropathy to warrant early and routine therapy in all Pima Indians with type 2 diabetes mellitus. DESIGN: A computerized medical decision analysis model was used to compare strategy 1, screening for microalbuminuria and treatment of incipient nephropathy as currently recommended with ACE inhibitor therapy, with strategy 2, a protocol wherein all patients were routinely administered an ACE inhibitor 1 year after diagnosis of type 2 diabetes mellitus. The model assumed that ACE inhibitors can block, at least in part, the pathogenic mechanisms responsible for early diabetic nephropathy (microalbuminuria). RESULTS: The model predicted that strategy 2 would produce more life-years at less cost than strategy 1, if routine drug therapy reduced the rate of development of microalbuminuria by 21% in all patients. Only a 9% reduction in the rate of development of microalbuminuria was cost-effective at $15,000 per additional life-year gained, and only a 2.4% reduction was cost-effective at $75,000 per additional life-year gained for strategy 2 over strategy 1. CONCLUSIONS: Routine ACE inhibitor therapy in Pima Indians with type 2 diabetes mellitus could prove more effective and even cost saving than the currently recommended approach of microalbuminuria screening. A prospective trial examining this goal should be considered. PMID- 10377931 TI - Cancer chemotherapy-related thrombotic thrombocytopenic purpura: biological evidence of increased nitric oxide production. AB - The occurrence of thrombotic thrombocytopenic purpura (TTP) in cancer patients receiving chemotherapy has been well established; although this entity is rare, its clinical importance seems to be growing. We describe 3 cases of TTP developing in cancer patients receiving different chemotherapeutic regimens. Using a sensitive high-performance liquid chromatographic method, we evaluated the stable nitric oxide end products, nitrite and nitrate, in the plasma of these patients. Nitric oxide is one of the key components involved in maintaining the normal nonthrombogenicity of the vascular endothelium. In our 3 patients, we found increased nitrate titers that were substantially higher than those observed in patients with de novo TTP. The observed increased release of nitrate could be interpreted as the consequence of massive disruption of endothelial integrity, with consequent passive nitric oxide release in vivo, or an adaptive mechanism of the endothelium to compensate for diffuse microvascular occlusion. The 2 mechanisms may both be involved, but the normal titers of nitric oxide end products in de novo TTP suggest that the former mechanism is more important, at least in cancer chemotherapy-related TTP. PMID- 10377930 TI - The incidence and prevalence of myoclonus in Olmsted County, Minnesota. AB - OBJECTIVE: To study the incidence and prevalence of diagnosed myoclonus in Olmsted County, Minnesota. Little is known about the frequency and distribution of myoclonus in the general population. DESIGN: Descriptive study with case ascertainment through a records-linkage system. PATIENTS AND METHODS: We used a medical records-linkage system to identify all subjects whose records contained documentation of myoclonus or of diseases known to exhibit myoclonus. The records of all potential patients were reviewed by a neurologist, and only patients with pathologic and persistent myoclonus were included. Population denominators were derived from census data. RESULTS: The average annual incidence rate of pathologic and persistent myoclonus for 1976 through 1990 was 1.3 cases per 100,000 person-years. The rate increased with advancing age and was consistently higher in men. Symptomatic myoclonus was the most common type, followed by epileptic and essential myoclonus; dementing diseases were the most common cause of symptomatic myoclonus. The lifetime prevalence of myoclonus, as of January 1, 1990, was 8.6 cases per 100,000 population, and the prevalence increased with advancing age. CONCLUSION: Although our figures are probably under-estimated, they are the first attempt, to our knowledge, to measure myoclonus morbidity in the general population. We found clinical features and age and sex distributions different from those previously described in clinical series. PMID- 10377932 TI - Toxoplasma infection in systemic lupus erythematosus mimicking lupus cerebritis. AB - An opportunistic infection is a known, although under-diagnosed, complication of systemic lupus erythematosus (SLE). A 48-year-old woman with a recent diagnosis of SLE was admitted to the hospital because of a fever, confused state, and convulsive episode. Her symptoms were interpreted as being compatible with lupus cerebritis. Treatment with methylprednisolone resulted in a temporary improvement in the patient's condition. Nevertheless, during the next few weeks, her physical and mental condition deteriorated, and she died of massive pulmonary emboli. An autopsy revealed no signs of lupus cerebritis; however, disseminated cerebral toxoplasmosis was found. Cerebral toxoplasmosis is a rare complication of SLE that may be misdiagnosed as lupus cerebritis. PMID- 10377933 TI - Vertebral Aspergillus osteomyelitis and acute diskitis in patients with chronic obstructive pulmonary disease. AB - Aspergillus osteomyelitis of the spine with acute diskitis has been well documented in immunocompromised hosts but is rare in immunocompetent patients. Predisposing factors to infection are prolonged neutropenia, hematologic malignancies, chemotherapy, history of prior spinal trauma or surgery, allograft transplantation, or any condition requiring the use of long-term immunosuppressive agents or systemic corticosteroids. Patients with chronic obstructive pulmonary disease (COPD) treated with systemic corticosteroids for either long-term management or frequent exacerbations are at potential risk for such infections. Patients with severe COPD treated primarily with inhaled corticosteroids are considered immunocompetent. This report describes 2 cases of Aspergillus osteomyelitis with acute diskitis in apparently immunocompetent patients with COPD who, aside from brief courses of systemic corticosteroids, were using inhaled corticosteroid therapy. One patient was treated with intravenous amphotericin B alone, whereas the other received amphotericin B and underwent surgical debridement. Both have done well and were symptom free at 6 month follow-up. PMID- 10377934 TI - Transmyocardial and percutaneous myocardial revascularization: current and future role in the treatment of coronary artery disease. AB - Transmyocardial revascularization (TMR) is a new treatment modality under evaluation in patients with severely symptomatic, diffuse coronary artery disease, in whom the potential for medical or interventional management has been exhausted. Preliminary clinical trials show improved ischemic symptoms within the first 3 months in about 70% of TMR-treated patients. The original proposed mechanism of surgical or catheter-based TMR (percutaneous myocardial revascularization [PMR]) was that channels mediate direct blood flow between the left ventricular cavity and ischemic myocardium. However, several alternative explanations for the clinical success of TMR have recently been suggested, including improved perfusion by angiogenesis, an anesthetic effect by nerve destruction, and a potential placebo effect. This article reviews the clinical role of TMR/PMR, its possible pathophysiologic mechanisms, and its controversies. It provides an overview of the actual scientific and clinical status of TMR and details future directions. PMID- 10377935 TI - Surgery in the patient with liver disease. AB - Patients with liver disease often undergo surgery. With the increasing prevalence of liver disease and improved survival due to newer medications and treatments, a growing number of patients with liver disease will require preoperative assessment. Because of the multiple physiological roles of the liver, hepatic dysfunction places these patients at an increased risk of perioperative morbidity and mortality. The precise risks associated with specific liver diseases are poorly understood but are greater with increased impairment of hepatic function. Identifying preexisting problems that could be optimally and appropriately managed before surgery (e.g., coagulation status, intravascular volume, renal function, electrolytes, cardiovascular status, and nutrition) may reduce these risks and decrease mortality in patients with liver disease undergoing surgery. PMID- 10377936 TI - Stanley Cohen--Nobel laureate for growth factor. PMID- 10377937 TI - Selective estrogen receptor modulators and phytoestrogens: new therapies for the postmenopausal women. AB - Estrogen deficiency in the postmenopausal woman results in numerous symptomatic and asymptomatic manifestations, including vasomotor symptoms, osteoporosis, heart disease, bladder and vaginal symptoms, and cardiovascular disease. Estrogen replacement therapy is associated with amelioration of these problems but has attendant risks. A newer class of drugs, the selective estrogen receptor modulators, provides both estrogen agonist and antagonist properties, depending on the target tissue. This article discusses the mechanism by which selective estrogen receptor modulators may vary in their end-organ effects and reviews the clinical studies associated with these compounds. Phytoestrogens are widely used in the United States, but little information is available regarding their potential long-term effects. PMID- 10377938 TI - 82-year-old man with recurrent syncope. PMID- 10377939 TI - The macrolides: erythromycin, clarithromycin, and azithromycin. AB - In addition to erythromycin, macrolides now available in the United States include azithromycin and clarithromycin. These two new macrolides are more chemically stable and better tolerated than erythromycin, and they have a broader antimicrobial spectrum than erythromycin against Mycobacterium avium complex (MAC), Haemophilus influenzae, nontuberculous mycobacteria, and Chlamydia trachomatis. All three macrolides have excellent activity against the atypical respiratory pathogens (C. pneumoniae and Mycoplasma species) and the Legionella species. Azithromycin and clarithromycin have pharmacokinetics that allow shorter dosing schedules because of prolonged tissue levels. Both azithromycin and clarithromycin are active agents for MAC prophylaxis in patients with late-stage acquired immunodeficiency syndrome (AIDS), although azithromycin may be the preferable agent because of fewer drug-drug interactions. Clarithromycin is the most active MAC antimicrobial agent and should be part of any drug regimen for treating active MAC disease in patients with or without AIDS. Although both azithromycin and clarithromycin are well tolerated by children, azithromycin has the advantage of shorter treatment regimens and improved tolerance, potentially improving compliance in the treatment of respiratory tract and skin or soft tissue infections. Intravenously administered azithromycin has been approved for treatment of adults with mild to moderate community-acquired pneumonia or pelvic inflammatory diseases. An area of concern is the increasing macrolide resistance that is being reported with some of the common pathogens, particularly Streptococcus pneumoniae, group A streptococci, and H. influenzae. The emergence of macrolide resistance with these common pathogens may limit the clinical usefulness of this class of antimicrobial agents in the future. PMID- 10377940 TI - The challenge of becoming a distinguished clinician. PMID- 10377941 TI - The Carney Triad: a lesson in observation, creativity, and perseverance. PMID- 10377942 TI - Efficacy of antineoplastons A10 and AS2-1. PMID- 10377943 TI - Plant stanol esters and vitamin K. PMID- 10377944 TI - Ku, a DNA repair protein with multiple cellular functions? AB - The Ku protein binds to DNA ends and other types of discontinuity in double stranded DNA. It is a tightly associated heterodimer of approximately 70 kDa and approximately 80 kDa subunits that together with the approximately 470 kDa catalytic subunit, DNA-PKcs, form the DNA-dependent protein kinase. This enzyme is involved in repairing DNA double-strand breaks (DSBs) caused, for example, by physiological oxidation reactions, V(D)J recombination, ionizing radiation and certain chemotherapeutic drugs. The Ku-dependent repair process, called illegitimate recombination or nonhomologous end joining (NHEJ), appears to be the main DNA DSB repair mechanism in mammalian cells. Ku itself is probably involved in stabilizing broken DNA ends, bringing them together and preparing them for ligation. Ku also recruits DNA-PKcs to the DSB, activating its kinase function. Targeted disruption of the genes encoding Ku70 and Ku80 has identified significant differences between Ku-deficient mice and DNA-PKcs-deficient mice. Although all three gene products are clearly involved in repairing ionizing radiation-induced damage and in V(D)J recombination, Ku-knockout mice are small, and their cells fail to proliferate in culture and show signs of premature senescence. Recent findings have implicated yeast Ku in telomeric structure in addition to NHEJ. Some of the phenotypes of the Ku-knockout mice may indicate a similar role for Ku at mammalian telomeres. PMID- 10377945 TI - Immortalization and characterization of Nijmegen Breakage syndrome fibroblasts. AB - Nijmegen Breakage Syndrome (NBS) is a very rare autosomal recessive chromosomal instability disorder characterized by microcephaly, growth retardation, immunodeficiency and a high incidence of malignancies. Cells from NBS patients are hypersensitive to ionizing radiation (IR) and display radioresistant DNA synthesis (RDS). NBS is caused by mutations in the NBS1 gene on chromosome 8q21 encoding a protein called nibrin. This protein is a component of the hMre11/hRad50 protein complex, suggesting a defect in DNA double-strand break (DSB) repair and/or cell cycle checkpoint function in NBS cells. We established SV40 transformed, immortal NBS fibroblasts, from primary cells derived from a Polish patient, carrying the common founder mutation 657del5. Immortalized NBS cells, like primary cells, are X-ray sensitive (2-fold) and display RDS following IR. They show an increased sensitivity to bleomycin (3.5-fold), etoposide (2.5 fold), camptothecin (3-fold) and mitomycin C (1.5-fold), but normal sensitivity towards UV-C. Despite the clear hypersensitivity towards DSB-inducing agents, the overall rates of DSB-rejoining in NBS cells as measured by pulsed field gel electrophoresis were found to be very similar to those of wild type cells. This indicates that the X-ray sensitivity of NBS cells is not directly caused by an overt defect in DSB repair. PMID- 10377946 TI - Cisplatin DNA cross-links do not inhibit S-phase and cause only a G2/M arrest in Saccharomyces cerevisiae. AB - Cisplatin (CDDP) has been used as a DNA cross-linking agent to evaluate whether there is a specific cell cycle checkpoint response to such damage in Saccharomyces cerevisiae (S. cerevisiae). Fluorescent-activated cell sorting (FACS) analysis showed only a G2/M checkpoint, normal exit from G1 and progression through S-phase following alpha-factor arrest and CDDP treatment. Of the checkpoint mutants tested, rad9, rad17 and rad24, did not show increased sensitivity to CDDP compared to isogenic wild-type cells. However, other checkpoint mutants tested (mec1, mec3 and rad53) showed increased sensitivity to CDDP, as did controls with a defect in excision repair (rad1 and rad14) or a defect in recombination (rad51 and rad52). Thus, by survival and cell cycle kinetics, it appears that DNA cross-links do not inhibit entry into S-phase or slow DNA replication and that replication continues after cisplatin treatment in yeast. PMID- 10377947 TI - SOS-dependent A-->G transitions induced by hydroxyl radical generating system hypoxanthine/xanthine oxidase/Fe3+/EDTA are accompanied by the increase of Fapy adenine content in M13 mp18 phage DNA. AB - Gas chromatography/isotope dilution-mass spectrometry with selected ion monitoring (GC/IDMS-SIM) was used to measure oxidised bases in hypoxanthine/xanthine oxidase/Fe3+/EDTA modified ss M13 mp18 phage DNA. A dose dependent increase of oxidised bases content in DNA was observed with the biggest augmentation of FapyGua, thymine glycol and FapyAde. The amount of 8-OH-Gua was relatively high both in non-oxidised and oxidised DNA, and increased to the same extent as FapyAde and ThyGly. DNA oxidation caused a dramatic decrease in phage survival after transfection to E. coli. Survival was improved 2.8-fold after induction of the SOS system by UV irradiation of bacteria and mutation frequency of the lacZ gene in SOS conditions increased 7-fold over that in non-irradiated bacteria. Spectrum of mutations was different from those reported previously and mutations were distributed rather randomly within M13 lacZ sequence, which was in contrast to previous findings, where with non-chelated metal ions other types of mutations were found in several clusters. Thus, conditions of DNA oxidation and accessibility of metal ions for DNA bases might be important factors for generating different DNA damages and mutations. Major base substitutions found both in SOS-induced and non-induced E. coli but with higher mutation frequency in SOS-induced cells were C-->A (approximately 20-fold increase in SOS-conditions), G-->A (9-fold increase) and G-->C (4.5-fold increase). Very few G-->T transitions were found. A particularly large group of A-->G transitions appeared only in SOS induced bacteria and was accompanied by augmentation of FapyAde content in the phage DNA with undetectable 2-OH-Ade. It is then possible that imidazole ring opened adenine mimics guanine during DNA replication and pairs with cytosine yielding A-->G transitions in SOS-induced bacteria. PMID- 10377948 TI - Chelating of iron and copper alters properties of DNA in L5178Y cells, as revealed by the comet assay. AB - We have previously found different proportions of iron and copper in nuclei of two sublines of murine lymphoma L5178Y (LY) and proposed a model of chromatin organization with these metal ions at the DNA attachment sites. We now examine the effect of chelators, desferal (DFO, iron-specific) and neocupreine (NEO, copper-specific) on DNA of LY-R and LY-S cells, using the comet and micronuclei frequency tests. There is less copper and more iron in LY-R nuclei than in LY-S nuclei. Accordingly, the effect of NEO is more marked in LY-R than in LY-S cells and in both sublines it is expressed as enhanced tail moment (measure of DNA damage in the comet assay) and increased micronuclei frequency. On the contrary, the effect of DFO on the tail moment is less pronounced in LY-R than in LY-S cells. With increasing DFO concentrations, there is a gradual decrease in the tail moment values below the control level in LY-S cells. In LY-R cells the tail moment values initially increase, then gradually decrease, eventually falling below the control level. This points to a dramatic conformational change that masks the effect of DNA discontinuities. The presence of the latter is indicated by the increase in micronuclei frequency. These results support the postulated differential role of iron and copper ions in maintaining the higher order DNA structure in LY sublines. PMID- 10377949 TI - p53 gene mutations in neoplastic transformation of C3H 10T1/2 and severe combined immunodeficiency fibroblasts. AB - The relevance of p53 mutations to the neoplastic malignant transformation of rodent fibroblasts by genotoxic physical and chemical agents is not clear. In the present study, we investigated p53 mutations (in exons 5-8) in non-transformed and neoplastically transformed C3H 10T1/2 and severe combined immunodeficiency (SCID) cells. No p53 mutations were detected in 15 neoplastically transformed (two spontaneous, one 3-methylcholanthrene-induced, seven gamma-ray-induced and five 'hot particle'-induced) and two non-transformed 10T1/2 cells. Wild-type p53 gene was also detected in all non-transformed (immortalized) SCID cell lines analyzed (four lines) whereas all three neoplastically transformed (two spontaneous, one gamma-ray-induced) cell lines displayed missense mutations in the p53 gene. These mutations were all transitions: A > G in codon 123, G > A in codon 152, and C > T in codon 238. We conclude that mutation in the p53 gene appears to be an infrequent event in 10T1/2 cells regardless of the transforming agent, but a frequent event in the neoplastic transformation of immortalized SCID cells. Non-transformed SCID cells are deficient in repair of DNA double-strand breaks, and neoplastically transformed cells are assumed to be deficient as well. PMID- 10377950 TI - Syntheses of some polyunsaturated sulfur- and oxygen-containing fatty acids related to eicosapentaenoic and docosahexaenoic acids. AB - With the aim of enhancing selectively the beneficial biological effects of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) a number of polyunsaturated fatty acids containing sulfur or oxygen atoms in the chain has been synthesized starting from EPA and DHA, respectively. Oxidative degradation of these acids led to the corresponding aldehydes all-(Z)-3,6,9,12 pentadecatetraenal and all-(Z)-3,6,9,12,15-octadecapentaenal. Reactions with DBU converted these aldehydes quantitatively into the conjugated isomers (2E,6Z,9Z,12Z)-pentadecatetraenal and (2E,6Z,9Z,12Z,15Z)-octadecapentaenal, respectively. The four aldehydes were transformed by a sequence of reactions comprising reduction to the alcohols, halogenation and substitution with mercapto esters into the corresponding sulfur containing polyunsaturated fatty acid esters. The oxygen containing esters were prepared from the respective alcohol by boron trifluoride catalysed reaction with ethyl diazoacetate. PMID- 10377951 TI - Preparation and antimicrobial studies of acyclic sulfamates. AB - A series of acyclic sulfamates have been prepared and tested for antimicrobial activity. Thus, the oxysulfonyl isocyanates, ROSO2NCO (1a, R = 4-methoxyphenyl; 1b, R = phenyl; 1c, R = 4-chlorophenyl and 1d, R = 2,2,2-trifluoroethyl) have been prepared in 76-91% yield from chlorosulfonyl isocyanate. Treatment of 1a-d with glycidol gave the glycidyl carbamates 2a d. Internal cyclisation afforded the corresponding 4-hydroxymethyl-2-oxazolidinones 3a-d, which in turn were hydrolysed to give the free amino alcohols 4a-d. The yields were in the range 39 85%. A preliminary agar diffusion test of 2a-d, 3a-d, 4a-d indicated 2a-d and 3c to be possible antimicrobial agents. A more thorough analysis of these compounds revealed a minimum inhibition concentration (MIC) of 128 and 64 mg l-1 for glycidyl p-methoxyphenoxysulfonylcarbamate (2a) and glycidyl phenoxysulfonylcarbamate (2b) respectively, against Branhamella catarrhalis. PMID- 10377952 TI - Capped-porphyrin precursors. AB - In the crystalline state, 2-[3-(tosyloxy)propoxy]benzaldehyde [(I), C17H18O5S] exists in a U-shaped conformation. The benzaldehyde and toluene rings are nearly parallel. Crystals of 2-[2-(tosyloxy)ethoxy]-benzaldehyde occur with two habits. The X-ray structure determinations of these habits reveal an anhydrous form [(II), C16H16O5S] and a hemihydrated form [(III), C16H16O5S.0.5H2O]. In (III), a water molecule bridges two carbonyl functions [O6...O1 2.87 (1) A]. 1,2,4,5 Tetrakis{2-[2-(1,3-dioxolan-2-yl)-phenoxy]ethoxy} benzene [(IV), C50H54O16] was prepared by protecting the aldehyde function of (II) or (III) with ethylene glycol and reacting the resulting compound with 1,2,4,5-tetrahydroxybenzene. Compound (IV) has 1 symmetry. PMID- 10377953 TI - Regulation of cation transport pathways and glycolytic enzyme activity by alterations in red cell volume. AB - In the presence of NH4Cl and hypotonic solutions, Rana balcanica red cells respond by increasing their volume. The stimulation of cellular volume by hypotonicity is more rapid than that of NH4Cl, while the maximum value is less than that observed in the presence of NH4Cl. Depending on the cause of swelling, (nct uptake of NH4Cl or decrease in external osmolality) cells show specific responses. The NH4Cl treatment causes a significant increase in intracellular Na+, from 5.14 +/- 0.78 to 29.84 +/- 0.47 mmoles l-1 cell, while hypotonicity leads to a significant decrease of this cation, to 3.85 +/- 0.25 mmoles l-1 cell in relation to the control, after 30 min of incubation of Rana balcanica erythrocytes. In addition, amiloiride significantly reverses the NH4Cl effect with respect to intracellular Na+. Both treatments cause a significant K+ loss in comparison with controls. Two glycolytic enzymes glyceraldehyde phosphate dehydrogenase (GAPDH) and pyruvate kinase (PK) of Rana balcanica haemolysate were found to respond to the NH4Cl effect by significantly decreasing their activity. PMID- 10377954 TI - Effect of cerebrocrast on the lymphocyte blast transformation activity in normal and streptozotocin-induced diabetic rats. AB - Both IDDM and NIDDM are characterized by deviations in peripheral T and B lymphocyte count, Thelper: Tsuppressor ratio, as well as by impaired Tsuppressor function. These abnormalities may promote insulin antibody and other antibody production, contributing to overt diabetes mellitus development in early stage of the disease. In the present study we explored the effects of cerebrocrast(1, 4 dihydropyridine derivative) administration on Con A- and IL-2-stimulated tissue lymphocyte blast transformation activity and on the thymus and lymph node mass in normal and streptozotocin (STZ)-induced diabetic rats. It was established that cerebrocrast, administered four times at the doses of 0.05 and 0.5 mg kg-1, has long-term (up to 14 days) effects on the immune system and protects against the toxic effect of STZ in STZ-induced diabetic rats, preventing thymus and lymph node mass loss. We conclude that cerebrocrast administration leads to the increase in number and activity of Thelper and Tsuppressor lymphocytes. Glycolysis and DNA synthesis in these cells is augmented under the influence of cerebrocrast administration. We propose that the increase in lymphocyte suppressive activity caused by cerebrocrast administration may prevent the development of IDDM and NIDDM in patients with pre-diabetes, but in patients with early and overt diabetes mellitus the drug administration may prevent the overexpression of insulin antibodies and other antibodies. The effect of cerebrocrast on the de novo production of insulin and IL-2 receptors may be beneficial for IDDM and NIDDM patients. PMID- 10377955 TI - Effects of some 1,4-dihydropyridine Ca antagonists on the blast transformation of rat spleen lymphocytes. AB - Ca antagonists of different classes (verapamil, nifedipine, nicardipinc, diltiazem) in a concentration of 10(-5) M and higher are known to suppress Ca2+ transport into the lymphocyte cytosol, changing a normal response of lymphocytes to mitogens and antigens and so inhibiting their proliferation, as well as IL-2 induced cell proliferation, and their receptor expression on the surface of lymphocytes without cell cytotoxicity. In the present work we studied the effect of some 1, 4-dihydropyridines (DHP) such as nimodipine, nicardipine, nifedipine, niludipine, cerebrocrast, etaftoron, as well as metabolites of cerebrocrast: compounds 7 and 8, (four of the last were synthesized in the Latvian Institute of Organic Synthesis) on rat spleen isolated lymphocyte activation and proliferation in vitro following stimulation with the mitogens concanavalin A (Con A) and recombinant interleukin-2 (IL-2), insulin and insulin antibodies. Based on the experimental results we conclude that in low concentrations (10(-7) to 10(-9) M) the tested 1, 4-DHP Ca antagonists stimulated the process of rat spleen lymphocyte proliferation and DNA synthesis, especially cerebrocrast. It is proposed that these Ca antagonists, as well as causing a concentration decrease of Ca2+, also activated phosphodiesterase, which in its turn, suppressed cAMP accumulation in the lymphocytes and eventually increased Ca2+ ion transport in the cells. Cerebrocrast among all the studied DHP Ca antagonists was the most potent in studies of activation of the lymphocytes in the presence of Con A, IL-2 and insulin, which indicates the number of suppressor and helper lymphocytes and formation of insulin and interleukin receptors on their membrane surface. The increase in the lymphocytes suppressive activity produced by this compound effect can prevent diabetes mellitus types I and II at the stages of pre-diabetes, early and distant diabetes, from hyperexpression of insulin and its receptor antibodies. PMID- 10377956 TI - Effect of polydeoxyribonucleotides on human fibroblasts in primary culture. AB - The effects of a mixture of oligo- and polydeoxyribonucleotides (PDRN) on the growth and protein secretion of cultured human skin fibroblasts were investigated. Both intact and DNAase-digested PDRN stimulated cell proliferation to a similar extent. When cultured fibroblasts were incubated with radioactive amino acids in the presence of intact or digested PDRN the incorporation of the tracer into secreted proteins increased significantly. This stimulation appears to be specific for certain protein components, including fibronectin. These results are interpreted assuming that PDRN and the nucleotides and nucleosides resulting from its degradation, can act as signal transducers or, alternatively, can be internalized and utilized to provide purine and pyrimidine rings for the salvage pathways. PMID- 10377957 TI - Influence of dietary selenium and vitamin E on the levels of fatty acids in brain and liver tissues of lambs. AB - In this study, the effects of dietary vitamin E, selenium, and their combination on the levels of fatty acid composition of the brain and liver tissues were examined. In brain tissue, the amounts of most fatty acids increased in vitamin E, combination and selenium groups compared with control group values. While the proportions of myristic, pentadecanoic, palmitic, linoleic, and total saturated fatty acids were decreased in vitamin E, Se and combination groups, eicosapentaenoic, total unsaturated and MUFA were increased in the same groups. In addition, the proportions arachidonic, eicosapentaenoic, total unsaturated, omega 6 and MUFA in the combination group were higher than in the control group. In liver tissue, the amounts of myristic, pentadecanoic, palmitic, eicosedienoic, eicosapentaenoic, docosahexaenoic, omega 3 and PUFA were higher in the combination group than in the control group. Also the proportions of eicosapentaenoic, docosahexaenoic acids in supplemented groups were higher than those in the control group. We conclude that dietary vitamin E and selenium have an influence on the levels of fatty acids in the brain and liver. PMID- 10377958 TI - The effect of coculture with human smooth muscle cells on the proliferation, the IL-1 beta secretion, the PDGF production and tube formation of human aortic endothelial cells. AB - Endothelial cells (ECs) and smooth muscle cells (SMCs), which are the major component cells of blood vessels, produce various bioactive substances and communicate with each other through them. Although several studies of the interaction between ECs and SMCs have been reported, the effect of coculture with SMCs on ECs is still obscure. To clarify the interaction of ECs and SMCs, we examined the effect of coculture with SMCs on the proliferation, the IL-1 beta secretion, the PDGF production and tube formation of ECs, using the coculture model: transferable wells and collagen gel. IL-1 and PDGF are considered to be related to progression of atherosclerosis. Proliferation and tube formation of ECs are associated with repair of vessels. In the transferable well system coculture with SMCs stimulated the proliferation of ECs, and enhanced the IL-1 beta secretion of ECs and in the collagen gel system coculture with SMCs induced the tube formation of ECs, and appeared to enhance the PDGF production of ECs. In conclusion, the effect of coculture with SMCs on ECs has two conflicting aspects: progression of atheroscleosis and angiogenesis. These results suggest that an imbalance of their effects may lead to pathological events. PMID- 10377959 TI - Neutrality of amiodarone on the initiation and propagation of membrane lipid peroxidation. AB - Amiodarone is an iodinated benzofuran derivative largely used as an antiarrhythmic. Owing to the sensitivity of heart tissue to radicals, amiodarone was assayed for putative effects on lipid peroxidation studied in liposomes of soybean phosphatidylcholine and of bovine heart mitochondrial lipids used as model systems. Lipid peroxidations were initiated with Fe2+/ascorbic acid, and with peroxyl radicals generated from the azocompounds, AAPH and AMVN. These assays were carried out by following the quenching of the fluorescent probe cis parinaric acid and by monitoring oxygen consumption. It has been ascertained that amiodarone does not protect or potentiate significantly the lipid peroxidation both lipidic systems. To fully ascertain the neutral behaviour of amiodarone in the lipid peroxidation process, the degradation of phospholipid acyl chains has been checked by GLC. These data confirm that amiodarone does not protect or potentiate lipid peroxidation to a significant extent. It is concluded that the limited effects of amiodarone might be related only indirectly with the lipid peroxidation. It is possible that the drug causes limited conformational and biophysical alterations in membrane phospholipid bilayers that can affect the process of peroxidation. Therefore, it is concluded that the therapeutic effects and benefits as a heart antiarrhythmic agent are independent of lipid peroxidation processes. Furthermore, the interaction of the drug with lipid bilayers does not induce significant conformational perturbations that could significantly favour or depress the peroxidation process. PMID- 10377960 TI - Effects of intraperitoneally-administered vitamin E and selenium on the blood biochemical and haematological parameters in rats. AB - The aim of this work was to determine the role of intraperitoneally-administered vitamin E and selenium on the biochemical and haematological parameters in the blood of rats. Thirty-two adult male Wistar rats were used in this study. All rats were randomly divided into four groups. The first group was used as the control. The second group was intraperitoneally administered with vitamin E (+/-( )alpha-tocopheroryl acetate, 10 mg day-1), the third group with Se (Na2SeO3 0.2 mg over a day), and the fourth group with vitamin E and Se (vitamin E 10 mg + Na2SeO3 0.2 mg over a day). This administration was done for 5 weeks. Blood samples were taken from animals at the end of the dosage period and biochemical parameters in serum samples and haematological parameters in total blood were determined. The levels of total cholesterol (p < 0.01) and number of white blood cell (p < 0.001) in blood were significantly higher in the vitamin E group than in the control group. The levels of ALP, total cholesterol (p < 0.01) and number of white blood cells (p < 0.01) in blood were significantly higher in the selenium group than in the controls. The levels of glucose (p < 0.05), ALP (p < 0.01), total cholesterol (p < 0.001) and number of white blood cells (p < 0.01) were higher in the vitamin E and selenium combined group than in the controls. Other parameters considered within this trial (ALT, LDH, creatinine, albumin, total protein, amylase, creatine kinase, HDL triglycerides, total lipid, sodium, chloride, uric acids, red blood cell, haemoglobin, packed cell volume, MCV, MCH, MCHC) did not show statistically significant differences between the control and injected groups. The results indicated that blood glucose and total cholesterol levels, ALP activity and white blood cell counts were significantly increased by intraperitoneal administration of vitamin E and selenium in rats. PMID- 10377961 TI - Long-lifetime lipid rhenium metal-ligand complex for probing membrane dynamics on the microsecond timescale. AB - We report the luminescence and spectral properties of a phospholipid analogue containing a long-lifetime luminescent rhenium metal-ligand complex (MLC) covalently linked to the amino group of phosphatidyl ethanolamine. When incorporated into synthetic membranes, this lipid probe displays intensity decay times near 3 microseconds. Importantly, the probe displays highly polarized emission with a maximal fundamental anisotropy of 0.33. This probe is expected to have numerous applications for studies of microsecond diffusion and dynamics of membranes. PMID- 10377962 TI - Action of quinolinic and indolinonic aminoxyls as radical-scavenging antioxidants. AB - The kinetic studies on the actions of quinolinic and indolinonic aminoxyls in the oxidation of lipid peroxidation induced by free radicals were carried out to evaluate their antioxidant activity. These aminoxyls showed a similar reactivity toward peroxyl radical with alpha-tocopherol. The antioxidant efficacies of aminoxyls against oxidation of methyl linoleate in homogeneous solution were smaller than that of alpha-tocopherol. Hydroxylamine, a reduced form of aminoxyl, possessed a comparative antioxidant efficacy with alpha-tocopherol and was capable of suppressing the consumption of alpha-tocopherol. Aminoxyls showed more potent antioxidant activity than alpha-tocopherol against the oxidation of methyl linoleate micelles induced by peroxyl radical or by a combination of copper ion and hydrogen peroxide. These results suggest that quinolinic and indolinonic aminoxyls may act as potent antioxidants against lipid peroxidation, especially in the presence of a good reductant which reduces aminoxyl radicals to hydroxylamines. PMID- 10377963 TI - Sterol synthesis. Preparation and characterization of fluorinated and deuterated analogs of oxygenated derivatives of cholesterol. AB - Oxygenated sterols, including both autoxidation products and sterol metabolites, have many important biological activities. Identification and quantitation of oxysterols by chromatographic and spectroscopic methods is greatly facilitated by the availability of authentic standards, and deuterated and fluorinated analogs are valuable as internal standards for quantitation. We describe the preparation, purification and characterization of 43 oxygenated sterols, including the 4 beta hydroxy, 7 alpha-hydroxy, 7 beta-hydroxy, 7-keto, and 19-hydroxy derivatives of cholesterol and their analogs with 25,26,26,26,27,27,27-heptafluoro (F7) and 26,26,26,27,27,27-hexadeuterio (d6) substitution. The 7 alpha-hydroxy, 7 beta hydroxy, and 7-keto derivatives of (25R)-cholest-5-ene-3 beta, 26-diol (1d) and their 16,16-dideuterio analogs were also prepared. These d2-26-hydroxysterols and [16,16-2H2]-(25R)-cholest-5-ene-3 beta, 26-diol (1e) were synthesized from [16,16 2H2]-(25R)-cholest-5-ene-3 beta, 26-diol diacetate (2e), which can be prepared from diosgenin. The highly specific deuterium incorporation at C-16 in 1e and 2e should be useful in mass spectral analysis of 26-hydroxycholesterol samples by isotope dilution methods. The delta 5-3 beta, 7 alpha, 26- and delta 5-3 beta, 7 beta, 26-triols were regioselectively oxidized/isomerized to the corresponding delta 4-3-ketosteroids with cholesterol oxidase. Also described are 5,6 alpha epoxy-5 alpha-cholestan-3 beta-ol, its 5 beta,6 beta-isomer, cholestane-3 beta, 5 alpha,6 beta-triol, their F7 and d6 derivatives, and d3-25-hydroxycholesterol, which was prepared from 3 beta-acetoxy-27-norcholest-5-en-25-one (30). The 43 oxysterols and most synthetic intermediates were isolated in high purity and characterized by chromatographic and spectroscopic methods, including mass spectrometry and nuclear magnetic resonance (NMR) spectroscopy. Detailed mass spectral assignments are presented, and 1H NMR stereochemical assignments are derived for the C-19 protons of 19-hydroxysterols and for the side-chain protons of 30. PMID- 10377964 TI - Synthesis and intermembrane transfer of pyrene-labelled liponucleotides: ceramide phosphothymidines. AB - Phospholipid conjugates of 3'-azido-3'-deoxythymidine (AZT) show activity against human immunodeficiency virus (HIV) in vitro. Here we report on the synthesis and characterization of two pyrene containing conjugates: 2-N-(4-(pyren-1 yl)butanoyl)ceramide 5'-phosphothymidine (Pbs-Cer-P-T) (XII) and 2-N-(10-(pyren-1 yl)decanoyl)ceramide 5'-phosphothymidine (Pds-Cer-P-T) (XIII). These fluorescent labelled conjugates served as model compounds to study incorporation of sphingoliponucleotides into membranes. The complex compounds were prepared by condensation of 3'-acetylthymidine and labelled ceramides using the phosphite triester coupling procedure. UV absorption, fluorimetry as well as 1H-, 31P-, 13C NMR analyses were used for structure confirmation of the synthesized substances. When incorporated into small unilamellar 1-palmitoyl-2-oleoyl-glycerophosphatidyl choline (POPC) vesicles and incubated with unlabelled acceptor POPC vesicles, the compounds (XII) and (XIII) exhibited spontaneous transfer. Kinetic data suggest that transfer from donor to acceptor vesicles occurred via the intervening aqueous phase. The non-specific lipid transfer protein from bovine liver stimulated the transfer of Pds-Cer-P-T between phospholipid vesicles in a concentration dependent manner. PMID- 10377965 TI - Interaction of retinol with dipalmitoylphosphatidylcholine and their formation of small dispersed particles. AB - Stable aqueous dispersions of all-trans-retinol (vitamin A, VA) were obtained by sonication with dipalmitoylphos-phatidylcholine (DPPC) in the VA mole fraction range 0.1-0.7. In order to clarify the dispersal mechanism, the dispersed particles were characterized and the interaction between VA and DPPC was investigated using several physicochemical techniques. Dynamic light scattering measurements showed that the diameter of the dispersed particles was 50-70 nm. A limited amount of VA was incorporated into DPPC bilayer membranes (approximately 5 mol%). The trapped aqueous volume inside the particles was determined fluorometrically using the aqueous space marker calcein and the volume in the VA/DPPC particles was decreased markedly with the addition of VA into small unilamellar vesicles of DPPC. The decline in the fraction of vesicular particles was also confirmed by fluorescence quenching of N-dansylhexadecylamine in the DPPC membrane by the addition of the quencher CuSO4. These results indicate that the excess VA separated from the DPPC bilayers is stabilized as emulsion particles by the DPPC surface monolayer. PMID- 10377966 TI - Genotoxicity of organic extracts of airborne particles in somatic cells of Drosophila melanogaster. AB - Complex mixtures extracted from air filters exposed for 24 h in two sessions (27 July and 02 August 1991) and at two locations (Merced, downtown, and Pedregal de San Angel, south-west) in Mexico City were analysed. The organic extracts were from airborne particles equal or smaller than 10 microns (PM10), and from total suspended particles (TSP). These organic extracts were assayed in the somatic mutation and recombination test (SMART) in wings of Drosophila melanogaster using two different crosses as well as in the Salmonella/microsome assay using strain TA98 with and without S9 fraction. The presence of polycyclic aromatic hydrocarbons (PAH) in the extracts was determined by gas chromatography. The genotoxic activities observed in the two test systems were comparable with the indirect mutagens producing greater response than the direct mutagens. The quantities of particulate matter as well as the genotoxic activities were higher on 02 August than on 27 July 1991 for both locations. The amounts of airborne particles and the resulting genotoxic activities were higher at Merced than at Pedregal. In both biological systems, PM10 were more genotoxic than TSP. These results demonstrate the sensitivity of the Drosophila wing SMART-which is an in vivo eukaryotic genotoxicity assay-as a biological monitor of environmental pollution related to airborne particles. PMID- 10377967 TI - Microbial dechlorination of polychlorinated biphenyls in anaerobic sewage sludge. AB - The potential of a chlorophenol (CP)-adapted consortium to dechlorinate polychlorinated biphenyls (PCBs) in sewage sludge was investigated. Results show that dechlorination rates differed significantly depending on sludge source and PCB congener. Higher total solid concentrations in sewage sludge and higher concentrations of chlorine in PCB resulted in slower dechlorination rates. No significant difference was found for 2,3,4,5-CB dechlorination from pH 6.0 to pH 8.0; however, dechlorination did not occur at pH 9.0 during a 41-day incubation period. Results show that at concentrations of 1 to 10 mg/L, the higher the PCB concentration, the faster the dechlorination rate. In addition, dechlorination rates were in the following order: methanogenic conditions > sulfate-reducing conditions > denitrifying conditions. The addition of acetate, lactate, pyruvate, and ferric chloride decreased lag times and enhanced dechlorination; however, the addition of manganese dioxide had an inhibitory effect. Dechlorination rates were also enhanced by the addition of PCB congeners, including 2,3,4-CB, 2,3,4,5-CB and 2,3,4,5,6-CB in mixture. Overall results show that the CP-adapted consortium has the potential to enhance PCB dechlorination. The optimal dechlorination conditions presented in this paper may be used as a reference for feasibility studies of PCB removal from sludge. PMID- 10377969 TI - PCDDs and PCDFs in vehicle exhaust particles in Japan. AB - Vehicle exhaust particles from gasoline and diesel engine cars were analyzed for PCDDs and PCDFs. The congener patterns of PCDDs and PCDFs in exhaust particles were different between gasoline and diesel engine cars. Suspended particulate matter from electrostatic precipitator connected to a highway tunnel was also analyzed for PCDDs and PCDFs. The congener pattern of suspended particular matter was different from both of gasoline and diesel engine cars. Total amounts of PCDDs/PCDFs sum concentrations in gasoline, diesel and suspended particulate matter were 0.21, 0.87 and 26.0 ng/g, respectively. The I-TEQs levels in gasoline, diesel and suspended particulate matter were 4.2, 11 and 242 pg/g, respectively. PMID- 10377970 TI - Introduction to pyrolysis-capillary gas chromatography. AB - By breaking large molecules into characteristic smaller fragments, analytical pyrolysis extends the use of gas chromatography to the analysis of polymeric materials, including natural polymers such as cellulose as well as synthetics. An understanding of the chemistry involved permits interpretation of the information present in these molecular fragments, with application to polymer and copolymer microstructure in addition to routine identification and quality control. Whether microfurnace, Curie-point or resistively heated filament, the pyrolysis device must be interfaced efficiently to the gas chromatography to make use of the resolving power afforded by capillary columns. PMID- 10377971 TI - Applied gas chromatography coupled to isotope ratio mass spectrometry. AB - Compound-specific isotope analysis (CSIA) by isotope ratio mass spectrometry (IRMS) following on-line combustion (C) of compounds separated by gas chromatography (GC) is a relatively young analytical method. Due to its ability to measure isotope distribution at natural abundance level with great accuracy and high precision, GC-C-IRMS has increasingly become the method of choice in authenticity control of foodstuffs and determination of origin in archaeology, geochemistry, and environmental chemistry. In combination with stable isotope labelled compounds, GC-C-IRMS is also used more and more in biochemical and biomedical application as it offers a reliable and risk-free alternative to the use of radioactive tracers. The literature on these topics is reviewed from the advent of commercial GC-C-IRMS systems in 1990 up to the beginning of 1998. Demands on sample preparation and quality of GC separation for GC-C-IRMS are discussed also. PMID- 10377972 TI - On-line combination of aqueous-sample preparation and capillary gas chromatography. AB - An overview is presented of methods currently in use to combine the preparation of aqueous samples on-line with capillary gas chromatography. Two approaches can be distinguished: heartcut-orientated reversed-phase liquid chromatography-gas chromatography (GC) and analyte-isolation-orientated analyte extraction-GC. These approaches either use techniques in which water is directly introduced onto the GC column, or an indirect approach in which water is eliminated, i.e., by solid phase extraction, solid-phase microextraction or liquid-liquid extraction, prior to introduction of the analytes onto the GC column. The latter type of approach is much more successful and user friendly, and many applications have been reported. PMID- 10377973 TI - Multifocal lenses compensate for chromatic defocus in vertebrate eyes. AB - The focal length of the vertebrate eye is a function of wavelength, i.e. the eye suffers from longitudinal chromatic aberration. Chromatic defocus is a particularly severe problem in eyes with high light-gathering ability, since depth of field is small due to a pupillary opening that is large in relation to the focal length of the eye. Calculations show that in such eyes only a narrow spectral band of light can be in focus on the retina. For the major part of the visual spectrum, spatial resolution should be limited by the optics of the eye and far lower than the resolving power achievable by the retinal cone photoreceptor mosaic. To solve this problem, fishes with irises unresponsive to light have developed lenses with multiple focal lengths. Well-focused images are created at the wavelengths of maximum absorbance of all spectral cone types. Multifocal lenses also appear to be present in some terrestrial species. In eyes with mobile irises, multifocal lenses are correlated with pupil shapes that allow all zones of the lens, with different refractive powers, to participate in the imaging process, irrespective of the state of pupil constriction. PMID- 10377974 TI - Response characteristics of neurons in the medial geniculate body of the little brown bat to simple and temporally-patterned sounds. AB - We examined the auditory response properties of neurons in the medial geniculate body of unanesthetized little brown bats (Myotis lucifugus). The units' selectivities to stimulus frequency, amplitude and duration were not significantly different from those of neurons in the inferior colliculus (Condon et al. 1994), which provides the primary excitatory input to the medial geniculate body, or in the auditory cortex (Condon et al. 1997) which receives primary input from the medial geniculate body. However, in response to trains of unmodulated tone pulses, the upper cutoff frequency for time-locked discharges (64 +/- 46.9 pulses per second or pps) and the mean number of spikes per pulse (19.2 +/- 12.2 pps), were intermediate to those for the inferior colliculus and auditory cortex. Further, in response to amplitude-modulated pulse trains, medial geniculate body units displayed a degree of response facilitation that was intermediate to that of the inferior colliculus and auditory cortex inferior colliculus: 1.32 +/- 0.33; medial geniculate body: 1.75 +/- 0.26; auditory cortex: 2.52 +/- 0.96, P < 0.01). These data suggest that the representation of isolated tone pulses is not significantly altered along the colliculothalamo cortical axis, but that the fidelity of representation of temporally patterned signals progressively degrades along this axis. The degradation in response fidelity allows the system to better extract the salient feature in complex amplitude-modulated signals. PMID- 10377975 TI - Effects of retinal dopamine depletion on the growth of the fish eye. AB - We investigated the suitability of fishes as animal models to study the involvement of the retinal dopaminergic system in the visually guided control of eye growth (emmetropization). Advantages of such a model system are (i) that all dopaminergic cells in the retina can be destroyed without apparent damage to other neurons, (ii) simple optical design and short depth of field of the eye, and (iii) continuous growth throughout life. Depleting the retina of dopamine in Aequidens pulcher (Cichlidae) had no apparent effect on refractive state, since size and focal length of the eye were reduced by the same amount. Furthermore, imposed defocus was compensated at a normal rate in spite of the absence of retinal dopamine. In A. pulcher, the dopaminergic system of the retina trus appears not to have an essential role in emmetropization. Our results furthermore suggest that in eyes of more complicated optical design, manipulation of the retinal dopaminergic system may lead to unrelated effects indistinguishable from direct interference with emmetropization. A major disadvantage of the fish model was that refractive state of the eye could not be measured accurately in vivo with standard methods. PMID- 10377976 TI - Thresholds for masking responses to light in three strains of retinally degenerate mice. AB - Mutant mice with retinal degeneration (rd/rd) were given 1-h pulses of light of varying brightness at times of the night when they would normally be active. The mutant mice showed a significantly greater inhibition of locomotor activity to light (negative masking) than wildtype controls. Lack of impairment, or even enhancement of negative masking suggests that this response may depend on sparing in retinally degenerate mice of the same receptor type that mediates clock resetting, because synchronization of the circadian system is known to be unimpaired in these mutants. With very dim light pulses, mutants did not change their activity, but wildtypes actually became more active (positive masking). Positive and negative masking appear to depend on different sensory and central processes. PMID- 10377977 TI - Masking of locomotor activity in hamsters. AB - The inhibition of locomotion by light (masking) was investigated in Syrian hamsters. When 1-h pulses of light were presented in the early night, activity was strongly suppressed by irradiances of about 1 lx or greater. Ultradian light dark cycles were used as another way to study masking. Hamsters were unable to entrain to 3.5:3.5-h light-dark cycles, thus permitting the masking and the entraining effects of light to be distinguished. Light had greater suppressive effects on activity in home cages than on activity in novel running wheels. Moreover, in home cages activity remained very low for about 30 min after lights were turned off. Post-pulse suppression of activity was not simply a consequence of reduced running, as shown by experiments in which running was temporarily prevented by locking the wheels. A phase response curve for masking was obtained by placing hamsters in novel wheels for 3-h periods at various times throughout their circadian cycles, and then superimposing a 30-min light pulse. The suppressive effect of light was maximal around the onset of activity, which normally coincides with dusk in hamsters. This may have adaptive value in limiting foraging to the hours of darkness. PMID- 10377978 TI - Masking by light in hamsters with SCN lesions. AB - Inhibition of wheel running by light (masking) was investigated in Syrian hamsters with suprachiasmatic nucleus or sham lesions. Approximately 90% of the wheel revolutions made by hamsters with complete suprachiasmatic nucleus lesions, as judged by histology and power spectrum analysis of their wheel running, occurred during the dark phases of an ultradian light-dark cycle (3.5 h light, 3.5 h dark). This was demonstrated for two illumination levels (380 lx and 6 lx). Similar results were obtained with sham-operated animals. In further tests, the hamsters with lesions also retained a strong preference for the dark side of a box divided into dark and light sides. These results demonstrate that the suprachiasmatic nucleus is not necessary for masking by light or the preference for a dark over a light compartment. Evidently the direct effects of light can substitute for the endogenous control by the suprachiasmatic nucleus to maintain appropriate behaviour in time and space. PMID- 10377979 TI - Enhanced masking response to light in hamsters with IGL lesions. AB - Syrian hamsters with intergeniculate leaflet or sham lesions were given tests with a series of light pulses of gradually decreasing intensities. The light pulses were given early in the night, at zeitgeber time 14-15. The amount of wheel running during the pulses was compared to that in the same hour on a night with no light pulses. Hamsters with intergeniculate leaflet lesions showed a significantly greater suppression of their wheel running in response to light than the sham-lesioned animals. The lesioned animals also had larger negative phase angles of entrainment to the 14:10-h light-dark cycle than sham-operated controls. However, phase shifting in response to light pulses at either zeitgeber time 14 or 18 was not significantly altered by the lesions. Preferences for spending more time in a dark than a light area were not abolished by the lesions. It is concluded that the intergeniculate leaflet in the Syrian hamster cannot be of paramount importance for masking of locomotor activity by light but may play a modulating role. PMID- 10377980 TI - Biogenic amines and division of labor in honey bee colonies. AB - Brain levels of dopamine, serotonin, and octopamine were measured in relation to both age-related division of labor and inter-individual differences in task specialization independent of age in honey bee colonies. The only differences among similarly aged bees performing different tasks were significantly lower levels of dopamine in food storers than comb builders and significantly lower levels of octopamine in soldiers than foragers, but soldiers also were slightly younger than foragers. Differences associated with age-related division of labor were stronger. Older bees, notably foragers, had significantly higher levels of all three amines than did younger bees working in the hive. Using social manipulations to unlink chronological age and behavioral status, octopamine was found to exhibit the most robust association between behavior and amine level, independent of age. Octopamine levels were significantly lower in normal-age nurses versus precocious foragers and overage nurses versus normal-age foragers, but not different in reverted nurses versus reversion colony foragers. Dopamine levels were significantly lower in normal-age nurses versus precocious foragers, but higher in reverted nurses versus reversion colony foragers. Serotonin levels did not differ in any of these comparisons. These correlative results suggest that octopamine is involved in the regulation of age-related division of labor in honey bees. PMID- 10377981 TI - Biogenic amines and division of labor in honey bee colonies: behaviorally related changes in the antennal lobes and age-related changes in the mushroom bodies. AB - Levels of the biogenic amines dopamine, serotonin, and octopamine were measured in different brain regions of adult worker honey bees as a function of age related division of labor, using social manipulations to unlink age and behavioral state. In the antennal lobes, foragers had higher levels of all three amines than nurses, regardless of age. Differences were larger for octopamine than serotonin or dopamine. In the mushroom bodies, older bees had higher levels of all three amines than younger bees, regardless of behavioral state. These correlative results suggest that increases in octopamine in the antennal lobes may be particularly important in the control of age-related division of labor in honey bees. PMID- 10377982 TI - Chemosensory responses of Tetrahymena thermophila to CB2, a 24-amino-acid fragment of lysozyme. AB - While lysozyme is a depolarizing chemorepellent in Tetrahymena, the entire lysozyme molecule is not necessary to activate the lysozyme receptor. Reduced lysozyme was cut into three fragments by cyanogen bromide cleavage and the fragments (CB1, CB2 and CB3) were separated by HPLC. Behavioral bioassays showed that the carboxy-terminal 24-amino-acid fragment, which we call CB2, is 100 times more active than intact lysozyme as a chemorepellent. CB2 appears to activate the same receptor as lysozyme because behavioral cross-adaptation is seen between these two compounds and an antibody generated to the purified lysozyme receptor blocks responses to both lysozyme and CB2. This is further supported by the observation that neomycin, which is a competitive inhibitor of lysozyme binding, also inhibits CB2 responses. This inhibition may be due to the fact that neomycin is highly positively charged (+5 at pH 7.0) and CB2 has a net charge of +4 at pH 7.0. Intracellular electrophysiological recordings documented that CB2 elicits a transient, depolarizing receptor potential that is similar to the lysozyme induced depolarizations except they are much smaller. CB2 is a more potent and specific ligand for use in studies of the lysozyme receptor of Tetrahymena. PMID- 10377983 TI - Peripheral frequency mis-match in the primitive ensiferan Cyphoderris monstrosa (Orthoptera: Haglidae). AB - Peripheral auditory frequency tuning in the ensiferan insect Cyphoderris monstrosa (Orthoptera: Haglidae) was examined by comparing tympanal vibrations and primary auditory receptor responses. In this species there is a mis-match between the frequency of maximal auditory sensitivity and the frequency content of the species' acoustic signals. The mis-match is not a function of the mechanical properties of the tympanum, but is evident at the level of primary receptors. There are two classes of primary receptors: low-tuned and broadly tuned. Differences in the absolute sensitivity of the two receptor types at the male song frequency would allow the auditory system to discriminate intraspecific signals from sounds containing lower frequencies. Comparisons of tympanal and receptor tuning indicated that the sensitivity of the broadly tuned receptors did not differ from that of the tympanum, while low-tuned receptors had significantly narrower frequency tuning. The results suggest that the limited specialization for the encoding of intraspecific signals in the auditory system of C. monstrosa is a primitive rather than a degenerate condition. The limited specialization of C. monstrosa may reflect the evolutionary origin of communication-related hearing from a generalized precursor through the addition of peripheral adaptations (tympana, additional receptors) to enhance frequency sensitivity and discrimination. PMID- 10377984 TI - The role of iron in protozoan and fungal infectious diseases. AB - To survive and replicate in vertebrate hosts, protozoan and fungal invaders must be capable of securing host iron. Successful pathogens obtain the metal from either extraction of heme, binding of siderophilins, binding of siderophores, and/or iron pools within host cells. The actual strategy can vary with the availability of iron in the particular host milieu. As a corollary, hosts have developed an elaborate iron withholding defense system. Conditions that can compromise the system as well as procedures that can strengthen it are reviewed. PMID- 10377985 TI - Analysis of genomic G + C content, codon usage, initiator codon context and translation termination sites in Tetrahymena thermophila. AB - In recent years, the amount of molecular sequencing data from Tetrahymena thermophila has dramatically increased. We analyzed G + C content, codon usage, initiator codon context and stop codon sites in the extremely A + T rich genome of this ciliate. Average G + C content was 38% for protein coding regions, 21% for 5' non-coding sequences, 19% for 3' non-coding sequences, 15% for introns, 19% for micronuclear limited sequences and 17% for macronuclear retained sequences flanking micronuclear specific regions. The 75 available T. thermophila protein coding sequences favored codons ending in T and, where possible, avoided those with G in the third position. Highly expressed genes were relatively G + C rich and exhibited an extremely biased pattern of codon usage while developmentally regulated genes were more A + T-rich and showed less codon usage bias. Regions immediately preceding Tetrahymena translation initiator codons were generally A-rich. For the 60 stop codons examined, the frequency of G in the end + 1 site was much higher than expected whereas C never occupied this position. PMID- 10377986 TI - Reduced invasion of cultured cells pretreated with a monoclonal antibody elicited against refractile body antigens of avian coccidial sporozoites. AB - The effect of a monoclonal antibody (1209-C2) elicited against sporozoite refractile-body antigen on invasion of cultured baby hamster kidney cells by avian Eimeria species was examined in vitro. Pretreatment of sporozoites with 1209-C2 for 45 min before inoculation into cultures or simultaneous introduction of sporozoites and 1209-C2 into cultures had no significant effect on invasion. However, pretreatment of cultures for 45 min with 1209-C2 (also with media from other cloned 1209 cell lines) significantly inhibited invasion by sporozoites of Eimeria tenella and E. adenoeides. Pretreatment of cultures with 2 unrelated monoclonal antibodies with the same isotype as 1209-C2 did not inhibit invasion by E. tenella. There was a significant correlation between time of exposure of the cultures to 1209-C2 and invasion (r = -0.80924; p = 0.0001), with inhibition of invasion occurring after 20 min exposure, but not after 10 min. There was also a significant correlation between the titer of 1209-C2 and invasion (r = 0.62291; p = 0.0305). Monoclonal antibody 1209-C2 cross-reacted with epitopes of baby hamster kidney cells by both immunofluorescence and Western blot. The fluorescent labeling of the cells differed according to the fixative that was used. In formalin-fixed cultures labeled with 1209-C2, fluorescent foci were distributed over the entire cell; after methanol fixation, 1209-C2 reacted with only discrete foci in the nucleus. On Western blots of sporozoites, 1209-C2 reacted with antigens having molecular sizes of approximately 8, 17, 23, 30, and 45-60 kDa, and with several minor bands. On baby hamster kidney cells, the antibody reacted primarily with bands of approximately 30, 45-60, and slightly with other bands. The data suggest that interactions among similar molecules in the sporozoites and host cells may play a role in cellular invasion. PMID- 10377987 TI - Cyclic expression of a nuclear protein in a dinoflagellate. AB - Nuclei of the dinoflagellate Crypthecodinium cohnii strain Whd were isolated and nuclear proteins were extracted in three fractions, corresponding to the increasing affinity of these proteins to genomic DNA. One fraction contained two major bands (48- and 46-kDa) and antibodies specific to this fraction revealed two major bands by Western blot on nuclear extracts, corresponding to the 46- and 48-kDa bands. The 48-kDa protein was detected in G1 phase but not in M phase cells. An expression cDNA library of C. cohnii was screened with these antibodies, and two different open reading frames were isolated. Dinoflagellate nuclear associated protein (Dinap1), one of these coding sequences, was produced in E. coli and appeared to correspond to the 48-kDa nuclear protein. No homologue of this sequence was found in the data bases, but two regions were identified, one including two putative zinc finger repeats, and one coding for two potential W/W domains. The second coding sequence showed a low similarity to non-specific sterol carrier proteins. Immunocytolocalization with specific polyclonal antibodies to recombinant Dinap1 showed that the nucleus was immunoreactive only during the G1 phase: the nucleoplasm was immunostained, while chromosome cores and nuclear envelopes were negative. PMID- 10377988 TI - Description of Opisthonecta matiensis n. sp. (Protozoa, Ciliophora), a new peritrich ciliate from wastewater. AB - Opisthonecta matiensis n. sp. was isolated from the inlet water of a wastewater treatment plant near Madrid, Spain, and studied in vivo, with silver methods, and using electronic and indirect immunofluorescence microscopy. This new species shows an amphora-like cell shape and has a size of 45-73 microns (x 58.2) x 25-40 microns (x 31.3). The oral infraciliature is formed by one haplokinety, three polykineties, and a short row of kinetosomes (epistomial membrane). The aboral infraciliature is made up of the trochal band and the scopula. From the trochal band arise three fibrillar systems: oral fibers, aboral fibers, and oblique fibers. The myoneme system is composed of a delicate peristomial ring, longitudinal branched fibers that reach the trochal band and of radial fibers extending from the scopula to the trochal band. The silverline system consists of an average of 147 lines. This new species is separated from other known forms by its smaller size, the presence of one single vacuole, and its higher number of silverlines. PMID- 10377989 TI - Correlation between loss of a Mg2+ conductance and an adaptation defect in a mutant of Paramecium tetraurelia. AB - Paramecium tetraurelia responds to chronic KCl-induced depolarization by swimming backward, but the ciliate recovers within seconds and then undergoes a prolonged adaptation period during which sensitivity to external stimuli is altered radically. We examined the role of Mg2+ in this phenomenon, prompted by finding that mutations in the eccentric-A gene both suppressed a Mg(2+)-specific conductance and prevented adaptation. Adaptation of the wild type proceeded normally when extracellular Mg2+ was varied from 0-20 mM, however, suggesting that channel-mediated Mg2+ fluxes were not involved. In seeking alternative explanations for the eccentric mutant phenotype, we ascertained that there was an osmotic component to adaptation but that K(+)-induced depolarization was the primary stimulus. We also noted that wild-type and eccentric mutant cells depolarized by equivalent amounts in KCl, suggesting that the genetic lesion must lie downstream of membrane-potential change. We also examined whether the adaptation-induced behavioral changes and, indeed, the defect in eccentric might be explained in terms of Mg2+ and Na+ efflux during behavioral testing, but experimental observations failed to support this notion. Finally, we consider the possibility that eccentric gene mutation prevents adaptation by interfering with intracellular free Mg2+ homeostasis in Paramecium. PMID- 10377990 TI - Babesia canis canis, Babesia canis vogeli, Babesia canis rossi: differentiation of the three subspecies by a restriction fragment length polymorphism analysis on amplified small subunit ribosomal RNA genes. AB - The parasites Babesia canis and Babesia gibsoni (phylum Apicomplexa) are responsible for canine babesiosis throughout the world. Babesia canis was previously described as a group of three biologically different subspecies, namely B. canis canis, B. canis vogeli, and B. canis rossi. We report partial sequences of small subunit ribosomal RNA gene (ssu-rDNA) of each subspecies amplified in vitro with primers derived from a semi-conserved region of the ssu rDNA genes in other Babesia species. The polymerase chain reaction combined with a restriction fragment length polymorphism analysis, using HinfI and TaqI restriction enzymes, confirmed the separation of B. canis into three subspecies. These sequences were compared with previously published sequences of other Babesia species. A phylogenetic approach showed that the three subspecies of B. canis belong to the clade of Babesia species sensu stricto where B. canis canis clusters with B. canis rossi whereas B. canis vogeli might form a monophyletic group with the cluster B. divergens and B. odocoilei. Our results show that the three subspecies of B. canis can readily be differentiated at the molecular level and suggest that they might be considered as true species. PMID- 10377991 TI - Tobacco retinoblastoma-related protein phosphorylated by a distinct cyclin dependent kinase complex with Cdc2/cyclin D in vitro. AB - The retinoblastoma (Rb) protein was originally identified as a product of a tumour suppressor gene that plays a pivotal role in regulating both the cell cycle and differentiation in mammals. The growth-suppressive activity of Rb is regulated by phosphorylation with cyclin-dependent kinase (CDK), and inactivation of the Rb function is one of the critical steps for transition from the G1 to the S phase. We report here the cloning of a cDNA (NtRb1) from Nicotiana tabacum which encodes a Rb-related protein, and show that this gene is expressed in all the organs examined at the mRNA level. We have demonstrated that NtRb1 interacts with tobacco cyclin D by using yeast two-hybrid and in vitro binding assays. In mammals, cyclin D can assemble with CDK4 and CDK6, but not with Cdc2, to form active complexes. Surprisingly, tobacco cyclin D and Cdc2 proteins can form a complex in insect cells, which is able to phosphorylate tobacco Rb-related protein in vitro. Using immunoprecipitation with the anti-cyclin D anti-body, cyclin D can be found in a complex with Cdc2 in suspension-cultured tobacco BY-2 cells. These results suggest that the cdc2 gene modulates the cell cycle through the phosphorylation of Rb-related protein by forming an active complex with cyclin D in plants. PMID- 10377992 TI - Plants have a sensitive perception system for the most conserved domain of bacterial flagellin. AB - The flagellum is an important virulence factor for bacteria pathogenic to animals and plants. Here we demonstrate that plants have a highly sensitive chemoperception system for eubacterial flagellins, specifically targeted to the most highly conserved domain within its N terminus. Synthetic peptides comprising 15-22 amino acids of this domain acted as elicitors of defence responses at sub nanomolar concentrations in cells of tomato and several other plant species. Peptides comprising only the central 8 to 11 amino acids of the active domain had no elicitor activity but acted as specific, competitive inhibitors in tomato cells. These antagonists suppressed the plant's response to flagellin, crude bacterial extracts and living bacterial cells. Thus, plants have a highly sensitive and selective perception system for the flagellin of motile eubacteria. PMID- 10377993 TI - A single locus determines sensitivity to bacterial flagellin in Arabidopsis thaliana. AB - Peptides corresponding to the most conserved domain of eubacterial flagellin act as potent elicitors in cells of different plant species. In intact Arabidposis thaliana seedlings these peptides (flg22 and flg15) caused callose deposition, induction of genes coding for pathogenesis-related proteins and a strong inhibition of growth. Half-maximal growth inhibition occurred at peptide concentrations of approximately 100 nM. In contrast, peptides representing the corresponding flagellin domains of the plant-associated bacteria A. tumefaciens and R. meliloti were inactive even at concentrations of 10 microM. With the exception of Ws-0, all ecotypes of A. thaliana tested were sensitive to flg22. Crosses of Ws-0 with the sensitive ecotypes Col-0 and La-er, respectively, resulted in sensitive F1 seedlings. In the F2 generation of both crosses, sensitivity segregated as a single trait with markers of chromosome 5 and a ratio of 3:1. Dominance of the locus sensing flagellin, termed FLS-1, suggests that it encodes an element which is important for the perception of the flagellin signal. PMID- 10377994 TI - A role for glutathione transferases functioning as glutathione peroxidases in resistance to multiple herbicides in black-grass. AB - Black-grass (Alopecurus myosuroides) is a major weed of wheat in Europe, with several populations having acquired resistance to multiple herbicides of differing modes of action. As compared with herbicide-susceptible black-grass, populations showing herbicide cross-resistance contained greatly elevated levels of a specific type I glutathione transferase (GST), termed AmGST2, but similar levels of a type III GST termed AmGST1. Following cloning and expression of the respective cDNAs, AmGST2 differed from AmGST1 in showing limited activity in detoxifying herbicides but high activities as a glutathione peroxidase (GPOX) capable of reducing organic hydroperoxides. In contrast to AmGST2, other GPOXs were not enhanced in the herbicide-resistant populations. Treatment with a range of herbicides used to control grass weeds in wheat resulted in increased levels of hydroperoxides in herbicide-susceptible populations but not in herbicide resistant plants, consistent with AmGST2 functioning to prevent oxidative injury caused as a primary or secondary effect of herbicide action. Increased AmGST2 expression in black-grass was associated with partial tolerance to the peroxidizing herbicide paraquat. The selective enhancement of AmGST2 expression resulted from a constitutively high expression of the respective gene, which was activated in herbicide-susceptible black-grass in response to herbicide safeners, dehydration and chemical treatments imposing oxidative stress. Our results provide strong evidence that GSTs can contribute to resistance to multiple herbicides by playing a role in oxidative stress tolerance in addition to detoxifying herbicides by catalysing their conjugation with glutathione. PMID- 10377995 TI - The sax1 dwarf mutant of Arabidopsis thaliana shows altered sensitivity of growth responses to abscisic acid, auxin, gibberellins and ethylene and is partially rescued by exogenous brassinosteroid. AB - Genetic approaches using Arabidopsis thaliana aimed at the identification of mutations affecting events involved in auxin signalling have usually led to the isolation of auxin-resistant mutants. From a selection screen specifically developed to isolate auxin-hypersensitive mutants, one mutant line was selected for its increased sensitivity to auxin (x 2 to 3) for the root elongation response. The genetic analysis of sax1 (hypersensitive to abscisic acid and auxin) indicated that the mutant phenotype segregates as a single recessive Mendelian locus, mapping to the lower arm of chromosome 1. Sax1 seedlings grown in vitro showed a short curled primary root and small, round, dark-green cotyledons. In the greenhouse, adult sax1 plants were characterized by a dwarf phenotype, delayed development and reduced fertility. Further physiological characterization of sax1 seedlings revealed that the most striking trait was a large increase (x 40) in ABA-sensitivity of root elongation and, to a lesser extent, of ABA-induced stomatal closure; in other respects, hypocotyl elongation was resistant to gibberellins and ethylene. These alterations in hormone sensitivity in sax1 plants co-segregated with the dwarf phenotype suggesting that processes involved in cell elongation are modified. Treatment of mutant seedlings with an exogenous brassinosteroid partially rescued a wild-type size, suggesting that brassinosteroid biosynthesis might be affected in sax1 plants. Wild-type sensitivities to ABA, auxin and gibberellins were also restored in sax1 plants by exogenous application of brassinosteroid, illustrating the pivotal importance of the BR-related SAX1 gene. PMID- 10377996 TI - The sax1 mutation defines a new locus involved in the brassinosteroid biosynthesis pathway in Arabidopsis thaliana. AB - In this issue we described a dwarf mutant in Arabidopsis thaliana, sax1, which is affected in brassinosteroid biosynthesis. This primary defect is responsible for alterations in hormone sensitivity of sax1 plants characterized by the hypersensitivity of root elongation to abscisic acid and auxin and the insensitivity of hypocotyl growth to gibberellins and ethylene (Ephritikhine et al., 1999; Plant J. 18, 303-314). In this paper, we report the further characterization of the sax1 mutant aimed at identification of the mutated step in the brassinosteroid biosynthesis pathway. Rescue experiments with various intermediates of the pathway showed that the sax1 mutation alters a very early step catalyzing the oxidation and isomerization of 3 beta-hydroxyl, delta 5,6 precursors to 3-oxo, delta 4,5 steroids. The mapping of the mutation, the physiological properties of the mutant and the rescue experiments indicate that sax1 defines a new locus in the brassinosteroid biosynthesis pathway. The SAX1 protein is involved in brassinosteroid-dependent growth of seedlings in both light and dark conditions. PMID- 10377997 TI - Characterization of an Arabidopsis thaliana receptor-like protein kinase gene activated by oxidative stress and pathogen attack. AB - An Arabidopsis thaliana cDNA clone that encodes a putative receptor-like protein kinase gene (At-RLK3) was characterized. The deduced 667-amino acid protein consists of an amino-terminal signal sequence, an extracellular domain, a single transmembrane domain, and a cytoplasmic domain with characteristics of serine/threonine protein kinase. Because of the original features of its extracellular domain, the At-RLK3 protein is a member of a new class of receptor like protein kinases. The At-RLK3 gene is present as a single copy within the Arabidopsis genome and its transcripts are detected in root, stem, leaf and flower. In cultured cells, the At-RLK3 gene is activated upon oxidative stress and salicylic acid treatment. In plants, the gene appears to be differentially regulated during various plant-pathogen interactions: upon inoculation with strains of Pseudomonas syringae pv. tomato harboring or not, different avr genes, At-RLK3 transcripts accumulate transiently at similar levels during both compatible and incompatible interactions. This gene is, however, preferentially expressed during the incompatible interaction induced by the soil-borne vascular bacteria, Ralstonia solanacearum. The involvement of At-RLK3 in signal transduction pathways during pathogen attack is discussed. PMID- 10377998 TI - BELL1 and AGAMOUS genes promote ovule identity in Arabidopsis thaliana. AB - Molecular and genetic analyses have demonstrated that the Arabidopsis thaliana gene BELL1 (BEL1) is required for proper morphogenesis of the ovule integuments. Several lines of evidence suggest that BEL1 may act, at least in part, to repress the function of the organ identity gene AGAMOUS (AG) during ovule development. To study the relative roles of BEL1 and AG, plants homozygous for ag, bel1 or both were constructed in an ap2 mutant background where ovules form even in the absence of AG function. The loss of either BEL1 or AG led to a decrease in the number of mature ovules, accompanied by an increase in primordial outgrowths. These data suggest that BEL1 and AG gene products act early in ovule development in a partially redundant manner to direct ovule identity. Development of the abnormal integuments characteristic of the Bel1- mutant phenotype was found to be dependent on AG function. Finally, BEL1 appears to be required for embryo sac development independent of both other aspects of ovule morphogenesis and AG function. This study therefore suggests that both BEL1 and AG are required for several distinct aspects of ovule morphogenesis. PMID- 10377999 TI - Time-resolved studies of the excited-state dynamics of meso tetra(hydroxylphenyl)chlorin in solution. AB - Meso-tetra(hydroxyphenyl)chlorin (m-THPC) is a new photosensitizer developed for potential use in photodynamic therapy (PDT) for cancer treatment. In PDT, the accepted mechanism of tumor destruction involves the formation of excited singlet oxygen via intermolecular energy transfer from the excited triplet-state dye to the ground triplet-state oxygen. Femtosecond transient absorption measurements are reported here for the excited singlet state dynamics of m-THPC in solution. The observed early time kinetics were best fit using a triple exponential function with time constants of 350 fs, 80 ps and > or = 3.3 ns. The fastest decay (350 fs) was attributed to either internal conversion from S2 to S1 or vibrational relaxation in S2. Multichannel time-resolved absorption and emission spectroscopies were also used to characterize the excited singlet and triplet states of the dye on nanosecond to microsecond time scales at varying concentrations of oxygen. The nanosecond time-resolved absorption data were fit with a double exponential with time constants of 14 ns and 250 ns in ambient air, corresponding to lifetimes of the S1 and T1 states, respectively. The decay of the T1 state varied linearly with oxygen concentration, from which the intrinsic decay rate constant, ki, of 1.5 x 10(6) s-1 and the biomolecular collisional quenching constant, kc, of 1.7 x 10(9) M-1 s-1 were determined. The lifetime of the S1 state of 10 ns was confirmed by fluorescence measurements. It was found to be independent of oxygen concentration and longer than lifetimes of other photosensitizers. PMID- 10378000 TI - Analysis of phosphorescence decay in heterogeneous systems: consequences of finite excitation flash duration. AB - Analysis of phosphorescence lifetimes using the Stern-Volmer equation is a reliable means of determining quencher concentration for a uniform sample. Methods of analysis for heterogeneous systems are based on the assumption that the excitation is produced by a momentary flash. This condition is an idealization because a real flash has a finite duration and a complex time profile. In the case of a heterogeneous quencher concentration, an excitation flash produces different initial intensities and different times of peak intensity from compartments having different concentrations of quencher. We formulated a model to explore the effects of flash duration on the shape of the emission curve obtained from systems in which the heterogeneity is continuous. We developed mathematical models that can be used to recover fitting parameters of continuous distributions of reciprocal lifetimes approximated as rectangular or Gaussian distributions, or an arbitrary histogram. We also formulated a procedure to convert the distribution of reciprocal lifetimes into a volume distribution of quencher concentration. We found that (1) the Stern-Volmer ratio of phosphorescence intensities cannot be employed for interpretation of pulse phosphorometric data in terms of a volume distribution of quencher; (2) shortening the flash duration decreases the difference of initial intensities between compartments having high and low quencher concentration; (3) the parameters of the volume distribution of quencher concentration can be recovered correctly only after taking account of the difference in initial intensities; and (4) calibration of the initial intensities for a given fitting delay and flash function is necessary. PMID- 10378001 TI - Photophysics of hypericin and hypocrellin A in complex with subcellular components: interactions with human serum albumin. AB - Time-resolved fluorescence and absorption measurements are performed on hypericin complexed with human serum albumin, HSA (1:4, 1:1 and approximately 5:1 hypericin: HSA complexes). Detailed comparisons with hypocrellin A/HSA complexes (1:4 and 1:1) are made. Our results are consistent with the conclusions of previous studies indicating that hypericin binds to HSA by means of a specific hydrogen-bonded interaction between its carbonyl oxygen and the N1-H of the tryptophan residue in the IIA subdomain of HSA. (They also indicate that some hypericin binds nonspecifically to the surface of the protein.) A single exponential rotational diffusion time of 31 ns is measured for hypericin bound to HSA, indicating that it is very rigidly held. Energy transfer from the tryptophan residue of HSA to hypericin is very efficient and is characterized by a critical distance of 94 A, from which we estimate a time constant for energy transfer of approximately 3 x 10(-15) s. Although it is tightly bound to HSA, hypericin is still capable of executing excited-state intramolecular proton (or hydrogen atom) transfer in the approximately 5:1 complex, albeit to a lesser extent than when it is free in solution. It appears that the proton transfer process is completely impeded in the 1:1 complex. The implications of these results for hypericin (and hypocrellin A) are discussed in terms of the mechanism of intramolecular excited state proton transfer, the mode of binding to HSA and the light-induced antiviral and antitumor activity. PMID- 10378002 TI - Excited-state properties of thymidine and their relevance to its heterogeneous emission in double-stranded DNA. AB - Published results on synthetic polynucleotides point to T as the major emitting fluorophore in DNA. We have reported also that the bases of the nonalternating polynucleotide poly(dA).poly(dT), in which T was selectively excited, undergo large-amplitude motions on the picosecond-nanosecond time scales (S. Georghiou et al., Biophys. J. 70, 1909-1922, 1996). In that study, the fluorescence decay profile of the T bases of this polynucleotide was found to contain a number of components; these may be considered to be the result of the motions of the bases that give rise to a distribution of stacked geometries of varying rigidity as well as dispersion and polar interactions. Here, we report the results of a study that we have undertaken in order to test this hypothesis. To this effect, we have studied the photophysical properties of thymidine (1) in aqueous buffer and in a number of organic solvents and (2) in aqueous sucrose solutions of viscosity extending to 149 cP. The results suggest that the fluorescence quantum yield decreases with an increase in the polarizability of the solvent, whereas it increases with an increase in the solvent polarity (on the basis of the empirical parameter of solvent polarity ETN) or viscosity. These findings suggest the following for the photophysical properties of the T bases in DNA: (1) Base stacking results in two antagonistic effects, namely it causes a reduction in fluorescence as a result of dispersion interactions and an enhancement as a result of a reduction in the motions of the bases and (2) exposure of the bases to the aqueous environment results in fluorescence enhancement. PMID- 10378003 TI - Evidence for [2 + 2] photoreaction of alpha-methylene-gamma-butyrolactones with thymine: an explanation for chronic actinic dermatitis to sesquiterpene lactones? AB - The photoreactivity of isoalantolactone, a natural sesquiterpene lactone, toward thymine was studied. After 313 nm irradiation of a deoxygenated acetone solution of isoalantolactone (2 x 10(-3) M) and thymine (4 x 10(-3) M), two intermolecular [2 + 2] photoadducts, 3 and 4, were isolated with respective yields of 30% and 18%. The structures of these two photoadducts were determined by a combination of NMR experiments. Compound 3 was identified as a cis-syn-exo intermolecular [2 + 2] photoadduct involving the 5,6 double bond of thymine and the exomethylenic double bond of the lactone, while compound 4 was identified as an intermolecular [2 + 2] photoadduct involving the same bonds but with the cis-syn-endo conformation. This high photoreactivity of sesquiterpene lactones toward thymine could be an explanation of the progressive evolution of allergic contact dermatitis toward chronic actinic dermatitis. PMID- 10378004 TI - Ultraviolet-B-induced damage to Escherichia coli Fpg protein. AB - We investigated the effect of UVB light (290 < or = lambda < or = 320 nm) on the structure and enzymatic activities of Escherichia coli Fpg protein (2,6-diamino-4 hydroxy-5N-methylformamidopyrimidine-DNA glycosylase), a DNA repair enzyme containing a zinc finger motif and five chromophoric Trp residues. Irradiation with UVB light of air-saturated pH 7.4 buffered aqueous solutions of Fpg induces the formation of polymers as shown by sodium dodecyl sulfate polyacrylamide gel electrophoretic analysis. In argon-saturated solutions, polymer formation produces a precipitate. The polymerization quantum yield is 0.07 +/- 0.01 and 0.15 +/- 0.02 in air- and argon-saturated solutions, respectively. In the polymerized Fpg protein, second-derivative absorption spectroscopy indicates that three and one Trp residues are destroyed in air- and argon-saturated solutions, respectively. Polymers are devoid of all three activities of the Fpg protein, whereas the unpolymerized protein retains full activities. Matrix-assisted laser desorption/ionization experiments demonstrate that polymer formation is accompanied by the formation of short polypeptides containing the first 32 or 33 residues of the N-terminal domain. Theses polypeptides are most probably formed by the photolytic cleavage of Fpg protein induced by light absorption by the adjacent Trp-34 residue. PMID- 10378005 TI - Merocyanine 540 solubilized as an ion pair with cationic surfactant in nonpolar solvents: spectral and photochemical properties. AB - Merocyanine 540 (MC) is an anionic dye that is used to photopurge the bone marrow of leukemia cells. Under these conditions it is localized mostly in cell membranes, which may affect its photochemical reactivity. We investigated the photochemistry of MC dissolved as a hydrophobic ion pair with a hexadecyltrioctadecylammonium cation in cyclohexane, trimethylpentane and toluene as well as in propylene carbonate, CH3CN, C2H5OH and D2O. In organic solvents, the absorption and fluorescence spectra of MC were strongly red-shifted compared with aqueous solutions. The fluorescence was also more intense despite aggregation that occurred in some solvents. Aggregation strongly affects the spectral and photochemical properties of MC, especially in aliphatic hydrocarbons in which distinctive H-type aggregates are formed. Hydrophobic MC is a moderate photosensitizer of singlet molecular oxygen (1O2). The following values for 1O2 quantum yields were calculated based on 1O2 phosphorescence relative to 1O2 generation by Rose Bengal: approximately 0.12 in trimethylpenthane, approximately 0.13 in cyclohexane, 0.045 in EtOH, 0.039 in toluene, 0.007 in CH3CN and approximately 3 x 10(-4) in D2O. The H-aggregates of MC in cyclohexane and trimethylpentane are better 1O2 producers than monomeric MC. The above 1O2 quantum yields are corrected for self-quenching because MC is an efficient 1O2 quencher (17 x 10(7) M-1 s-1 in CH3CN, 6.8 x 10(7) M-1 s-1 in D2O, 5.2 x 10(7) M 1 s-1 in EtOH, and 1.4 x 10(7) M-1 s-1 in toluene). Merocyanine undergoes photodegradation, a solvent-dependent process that proceeds faster when the dye is aggregated. The initial photodegradation rate is much slower in organic solvents than in water, but photodegradation products accumulated during longer irradiation may increase the rate in most solvents. Higher photostability and better photosensitization by MC in hydrophobic nonpolar solvents suggest that the killing of leukemia cells via a photodynamic mechanism may operate mostly in cell membranes. In contrast, any cytotoxic products from photodecomposition may be important in hydrophilic cell compartments. Our data show the spectral and photochemical properties of MC in a pure hydrophobic environment. PMID- 10378006 TI - Photoionization of ferrocytochrome c by 248 nm laser light and the observation of the early stages of ferricytochrome c unfolding in the nanosecond-to-millisecond timescale. AB - Photolysis of ferrocytochrome c by 248 nm laser light in aqueous solution at pH 7 generates hydrated electrons (eaq-) by a monophotonic process with quantum yield phi = 0.034. Approximately three-quarters of the eaq- originate from the heme, which is converted from the ferrous to the ferric state in < 100 ns. The conformational changes associated with the change in the redox state of cytochrome c are either not detectable spectrophotometrically or complete in < 100 ns. Also, under conditions where ferrocytochrome c is stable but ferricytochrome c is unfolded (3 M guanidine, pH 7, 40 degrees C), photoionization of ferrocytochrome c generated ferricytochrome c with similar quantum yield. Under these conditions, the lifetime of native ferricytochrome c is 67 microseconds; it decays via two intermediates with lambda max > 410 nm, neither of which is the thermodynamically favored, unfolded form. These species are putatively identified as unfolding intermediates with nonnative iron ligands, similar to those found during folding of ferrocytochrome c. The results suggest that unfolding, like folding, proceeds by intrachain diffusion and ligand exchange. PMID- 10378007 TI - The human melanocyte as a particular target for UVA radiation and an endpoint for photoprotection assessment. AB - The induction of DNA breaks by UVA (320-400 nm) in the nucleus of normal human melanocytes in culture was investigated using single cell gel electrophoresis, also called the comet assay. Endogenous pigment and/or melanin-related molecules were found to enhance DNA breakage: comets were more intense in melanocytes than in fibroblasts, in cells with high melanin content or after stimulation of melanogenesis by supplying tyrosine in the culture medium. After UVA doses where strong comets were observed, neither cytotoxicity nor stimulation of tyrosinase activity were detected. However, the accumulation of p53 protein suggested that cells reacted to genotoxic stress under these experimental conditions. The same approach was used to compare two sunscreens with identical sun protection factors but different UVA protection factors. The results presented in this paper suggest that human melanocytes may be used as a target cell to evidence broadspectrum photoprotection. Moreover, these data appear to be helpful in getting a better understanding of the role of sunlight in the initiating steps of melanocyte transformation. PMID- 10378008 TI - Differences in sensitivity to UVC, UVB and UVA radiation of a multidrug-resistant cell line overexpressing P-glycoprotein. AB - Multidrug resistance (MDR) is the phenomenon in which cultured tumor cells, selected for resistance to one chemotherapeutic agent, simultaneously acquire resistance to several apparently unrelated drugs. The MDR phenotype is multifactorial. The best-studied mechanism involves the expression of a membrane protein that acts as an energy-dependent efflux pump, known as P-glycoprotein (Pgp), capable of extruding toxic materials from the cell. In this work, resistance to UVA radiation, but not to UVC nor UVB, was observed in an MDR leukemia cell line. This cell line overexpresses Pgp. To study the role of Pgp in the resistance to UVA radiation, two MDR modulators or reversing agents (verapamil and cyclosporin A) capable of blocking Pgp activity were used. Cell viability was assessed and the techniques of flow cytometry and fluorescence microscopy were employed to measure the extrusion of rhodamine 123 by the efflux pump. The results show that MDR modulators did not modify the resistance to UVA radiation. Furthermore, although cell viability was not significantly altered, Pgp function was impaired after UVA treatment, suggesting that this glycoprotein may be a physical target for oxidative damage, and that other factors may be responsible for the UVA resistance. In agreement with this, it was found that the resistant cell line presented a higher catalase activity than the parental (non MDR) cell line. PMID- 10378009 TI - Photoproduct formation from a zinc benzochlorin iminium salt detected by fluorescence microscopy. AB - When examined by fluorescence microscopy, tumor cells loaded with a zinc benzochlorin iminium salt showed a very faint deep-red fluorescence that was rapidly transformed to a substantially brighter red-orange fluorescence. Fourier transfer spectroscopy analysis with a red-sensitive detection system revealed that an initial fluorescence at 770 nm was gradually converted to 640 nm fluorescence during excitation. Image analysis showed that photoproduct formation was accompanied by a change in the site of drug localization from the cytoplasm to the nucleus. These studies illustrate the power of interferometry for the characterization of photoproducts and changes in sensitizer localization during photoproduct formation. PMID- 10378010 TI - Enhancement of ALA-PDT damage by IR-induced hyperthermia on a colon carcinoma model. AB - The interaction of photodynamic therapy (PDT) with 5-aminolevulinic acid (ALA) and hyperthermia is not well understood. In the present study, significant enhancement of tumor damage was observed after simultaneous application of ALA PDT and IR-induced hyperthermia using a broad-band incoherent light source. One hour after systemic administration of ALA at a dose of 200 mg/kg, subcutaneously transplanted C26 colon carcinoma tumors were irradiated with two bands of the VersaLight system, red (R, 580-720 nm) and red plus IR (R + IR, 580-720 nm and 1250-1600 nm). Photoirradiation using the R + IR band at different fluence rates and exposures caused mild heating of the tumor to 39-43 degrees C at a 3 mm depth. Electron microscopy after ALA + R, ALA + R + IR and R + IR treatments showed early mitochondrial swelling that was more pronounced in the ALA + R + IR group. Tumor necrosis assessment, using histology and vital staining, revealed an enhancement of tumor necrosis depth in the ALA + R + IR group compared to ALA + R and R + IR. The results showed that subhyperthermic heating to 39-39.5 degrees C in the ALA + R + IR group decreased the threshold light dose required for 100% tumor necrosis from 210 J/cm2 (observed in the ALA + R group) to 140 J/cm2. A tumor growth delay test, based on tumor volume measurement, also revealed significant enhancement of antitumor effect after application of ALA + R + IR compared to ALA + R. PMID- 10378011 TI - Irradiation of amelanotic melanoma cells with 532 nm high peak power pulsed laser radiation in the presence of the photothermal sensitizer Cu(II)-hematoporphyrin: a new approach to cell photoinactivation. AB - Cu(II)-hematoporphyrin (CuHp) was efficiently accumulated by B78H1 amelanotic melanoma cells upon incubation with porphyrin concentrations up to 52 microM. When the cells incubated for 18 h with 13 microM CuHp were irradiated with 532 nm light from a Q-switched Nd: YAG laser operated in a pulsed mode (10 ns pulses, 10 Hz) a significant decrease in cell survival was observed. The cell photoinactivation was not the consequence of a photodynamic process, as CuHp gave no detectable triplet signal upon laser flash photolysis excitation and no decrease in cell survival was observed upon continuous wave irradiation. Thus, it is likely that CuHp sensitization takes place by photothermal pathways. The efficiency of the photoprocess was modulated by different parameters; thus, while varying the amount of added CuHp in the 3.25-26 microM range had little effect, pulse energies larger than 50 mJ and irradiation times of at least 120 s were necessary to induce a cell inactivation of about 50%. The porphyrin-cell incubation time prior to irradiation had a major influence on cell survival, suggesting that the nature of the CuHp microenvironment can control the efficiency of photothermal sensitization. PMID- 10378012 TI - Photobleaching of arterial fluorescent compounds: characterization of elastin, collagen and cholesterol time-resolved spectra during prolonged ultraviolet irradiation. AB - To study the photobleaching of the main fluorescent compounds of the arterial wall, we repeatedly measured the time-resolved fluorescence of elastin, collagen and cholesterol during 560 s of excitation with nitrogen laser pulses. Three fluence rate levels were used: 0.72, 7.25 and 21.75 microW/mm2. The irradiation related changes of the fluorescence intensity and of the time-resolved fluorescence decay constants were characterized for the emission at 390, 430 and 470 nm. The fluorescence intensity at 390 nm decreased by 25-35% when the fluence delivered was 4 mJ/mm2, a common value in fluorescence studies of the arterial wall. Cholesterol fluorescence photobleached the most, and elastin fluorescence photobleached the least. Photobleaching was most intense at 390 nm and least intense at 470 nm such that the emission spectra of the three compounds were markedly distorted by photobleaching. The time-resolved decay constants and the fluorescence lifetime were not altered by irradiation when the fluence was below 4 mJ/mm2. The spectral distortions associated with photobleaching complicate the interpretation of arterial wall fluorescence in terms of tissue content in elastin, collagen and cholesterol. Use of the time-dependent features of the emission that are not altered by photobleaching should increase the accuracy of arterial wall analysis by fluorescence spectroscopy. PMID- 10378013 TI - Knowing in the context of acting: the task dynamics of the A-not-B error. AB - The A-not-B error is one of the most robust and highly studied phenomena in developmental psychology. The traditional Piagetian interpretation is that the error reflects the immaturity of infants' understanding of objects as permanent entities. More recently, the error has been interpreted in terms of changes in representation, in memory, in spatial knowledge, and in inhibitory processes. Each account may be partially right but none offers a unified account of the many accumulated facts about this error. This article presents and tests a new unified explanation. The authors propose that the perseverative reach back to A is the product of the processes that take a hand to a location in visual space: the body centered nature of the spatial code, memories for previous reaching activity, and the close coupling of looking and reaching. The results from 6 experiments support this explanation. The results are used to challenge the idea of knowledge independent of and distinct from behavior. PMID- 10378014 TI - Connectionist and diffusion models of reaction time. AB - Two connectionist frameworks, GRAIN (J. L. McClelland, 1993) and brain-state-in-a box (J. A. Anderson, 1991), and R. Ratcliff's (1978) diffusion model were evaluated using data from a signal detection task. Dependent variables included response probabilities, reaction times for correct and error responses, and shapes of reaction-time distributions. The diffusion model accounted for all aspects of the data, including error reaction times that had previously been a problem for all response-time models. The connectionist models accounted for many aspects of the data adequately, but each failed to a greater or lesser degree in important ways except for one model that was similar to the diffusion model. The findings advance the development of the diffusion model and show that the long tradition of reaction-time research and theory is a fertile domain for development and testing of connectionist assumptions about how decisions are generated over time. PMID- 10378015 TI - The biological affects: a typology. AB - This typology of biological affects is based on developmental-interactionist theory of motivation, emotion, and cognition. Affects--subjectively experienced feelings and desires--involve interoceptive perceptual systems based on primordial molecules that characterize neurochemicals. Biological affects involve primary motivational-emotional systems (primes) associated with hierarchically organized neurochemical systems in the brain, including subcortical (reptilian) and paleocortical (limbic) brain structures. Affects fulfill individualistic (selfish) functions (arousal, approach-avoidance, agonistic) and prosocial (cooperative) functions. Selfish and cooperative functions are associated respectively with the right and left hemispheres. Biological affects constitute the physiological bases for higher level affects: social affects (e.g., pride, guilt, shame, pity, jealousy), cognitive affects (e.g., curiosity, surprise), and moral affects. PMID- 10378016 TI - Medical care in the next century. PMID- 10378017 TI - Prinzmetal's variant angina: three case reports and a review of the literature. AB - Prinzmetal's variant angina is a rare entity. When angina-like symptoms occur at rest, mostly at a specific hour in the early morning, together with transient ST segment elevations and angiographically normal arteries, provocative tests with ergonovine or acetylcholine should be performed. Endothelial dysfunction, a strong thrombotic tendency, an increased platelet aggregation together with changes in autonomic tone can trigger coronary vasospasms. Once treated with calcium antagonists and nitrates the prognosis is excellent and severe complications such as arrhythmias, myocardial infarction or sudden death are extremely rare. Coronary stenting can be useful for refractory coronary spasm, CABG can be used for important coronary atherosclerosis. This review is illustrated with three typical presentations of variant angina: a myocardial infarction without significant organic coronary atherosclerosis, an ergonovine induced coronary spasm with a non-significant coronary lesion and a multivessel spasm complicated by ventricular arrhythmia. All these three patients became asymptomatic after a treatment with calcium antagonists and nitrates. PMID- 10378018 TI - Plasma lipoprotein (a) concentrations in hypothyroid, euthyroid and hyperthyroid subjects. AB - OBJECTIVE: Alterations of the lipid profile are a well known phenomenon in thyroid dysfunction. Thyroid hormones regulate lipid metabolism through various mechanisms, but a key role is played by the LDL receptor pathway. Thyroid hormone influence on lipoprotein (a) [Lp(a)] metabolism is known. METHODS AND RESULTS: Therefore we studied Lp(a) concentrations in a group of 16 hypothyroid patients and in a group of 22 hyperthyroid patients. Twenty-six euthyroid subjects were used as a control group. Plasma Lp(a) concentrations in hyperthyroid patients (23.2 +/- 28.1 mg/dl) were significantly lower than those of the hypothyroid patients (27.1 +/- 19.2, p < 0.05). There were negative correlations between plasma Lp(a) concentrations and total T4 levels in patients with hyperthyroidism and hypothyroidism (r: -0.49, p < 0.05; r: -0.40, p < 0.05, respectively). Also, decreased HDL-C levels, increased LDL-C, total cholesterol and apo B levels in the hypothyroid patients according to euthyroid subjects were observed (p < 0.05). Decreased LDL-C levels, increased HDL-C and apo Al levels in the hyperthyroid patients according to euthyroid subjects were determined (p < 0.05). CONCLUSIONS: It was concluded that plasma Lp(a) concentrations increase in hypothyroid patients and the observed relationships between thyroid status and Lp(a) levels can be explained by impaired catabolism of apo B and Lp(a) in hypothyroidism. PMID- 10378019 TI - Adenoviruses and enteroviruses as pathogens in myocarditis and dilated cardiomyopathy. AB - BACKGROUND: Enteroviruses were detected in up to 50% in myocardium of patients with myocarditis and dilated cardiomyopathy, the latter being considered as a result of a prior subclinical myocarditis. A wide range of other infectious agents are being discussed as pathogens, often only based on reports of single cases. Adenovirus genome was recently identified in a significant number in the myocardium of paediatric patients with myocarditis. However, data on the role of adenoviruses for the aetiopathogenesis of myocarditis in adult patients is missing so far. Therefore, we studied the prevalence of adenoviral and enteroviral genome in myocardium of adults with myocarditis and dilated cardiomyopathy. METHODS: 15 patients were diagnosed at baseline with myocarditis, 16 patients with dilated cardiomyopathy according to clinical and histological criteria. Endomyocardial biopsies of these patients and 8 control patients with non-infectious heart diseases were evaluated by polymerase chain reactions for enterovirus and adenovirus genome. RESULTS: Enteroviral genome was detected in 27.3% patients with myocarditis or dilated cardiomyopathy, whereas adenoviral genome was not identified in any patient. Samples from control subjects systematically yielded negative results. CONCLUSIONS: From our data, it seems doubtful that adenoviruses are major pathogens of myocarditis or DCM, whereas enterovirus genome was identified in a significant number of patients with both diseases. PMID- 10378020 TI - Recurrent syncopal episodes of neurocardiogenic origin in a patient suffering from cardiac syndrome-X. AB - We describe the case of a 44-year-old man, with a history of recurrent syncopal episodes and effort angina, the latter attributed to cardiac syndrome-X, who was admitted to our department because of a syncopal episode. During his hospitalization laboratory investigations including haematologic and blood chemical findings, head C/T scan, electroencephalogram, 48-hour Holter monitoring, electrophysiologic testing and echocardiographic study disclosed no abnormalities. On the contrary, a passive upright tilt testing was found to be positive, resulting, approximately, in a 10-seconds time interval of asystole accompanied by syncope. The association in the same patient of cardiac syndrome-X and neurocardiogenic syncope, although never described before, might be explained by a similar pathophysiological mechanism, which is a sympathovagal imbalance. PMID- 10378021 TI - Anomalous origin of a coronary artery in a transplanted heart. AB - Upon routine coronary angiography one year after surgery in a 62-year-old male recipient of a heart transplant, an abnormal origin of the left anterior descending coronary artery from the pulmonary artery was found in the donor heart. This very rare congenital anomaly had not been detected during harvesting and transplantation of the heart, and to our knowledge it has never been described before in a heart transplant patient. The donor was a 43-year-old male who died of a spontaneous intracranial bleeding. The recipient continues to enjoy a normal functional capacity and is free of anginal complaints, though there is evidence of ischaemia in the left anterior descending artery territory on exercise thallium-201 myocardial perfusion imaging. PMID- 10378022 TI - Recurrent pulmonary emboli and thrombus attached to a permanently implanted pacemaker wire in pregnancy. AB - We present a case of pacer wire thrombus and recurrent pulmonary emboli in pregnancy associated with a permanent pacemaker. Transthoracic echocardiography demonstrated a thrombus attached to the pacer wire at the point where it crossed the tricuspid valve. After the uncomplicated vaginal delivery, thrombolytic therapy was given. This thrombus persisted despite thrombolytic therapy. Consequently, the patient was referred for cardiac surgery. The suspected cause was confirmed during the surgery. PMID- 10378023 TI - Paroxysmal atrioventricular block induced during head-up tilt test. AB - A 71-year-old female with vasovagal near-syncope suffered from paroxysmal second degree AV block during Holter monitoring. AV block was easily reproduced during head-up tilt test. She was successfully treated with a dual chamber pacemaker. This treatment is unusual and the role of cardiac pacing in patients with vasovagal symptoms is reviewed. PMID- 10378024 TI - Increased T-helper-1-type immunity and decreased T-helper-2-type immunity in patients with preeclampsia. AB - PROBLEM: To examine whether preeclampsia involves type-1 T-helper (TH1) immune hyperactivity. METHOD OF STUDY: Expression of HLA-DR, a cell-surface marker of activation, was analyzed on CD3+, CD4+, and CD8+ T cells in 15 preeclamptic patients and 15 normal pregnant women using flow cytometry. Additionally, peripheral blood mononuclear cells from preeclamptic patients and normal pregnant women were cultured with or without phytohemagglutinin (PHA) stimulation, and interleukin (IL)-2, IL-4, interferon (IFN)-gamma, and tumor necrosis factor (TNF) alpha concentrations were determined in the supernatant by immunoassays. RESULTS: HLA-DR antigen was expressed more strongly on CD3+ T cells in preeclamptic patients than in normal subjects. In preeclampsia, HLA-DR was expressed more strongly in CD8+ T cells than in CD4- T cells. More TNF-alpha, IL-2, and IFN gamma were produced by unstimulated and stimulated cultured peripheral blood mononuclear cells from preeclampsia patients than by those from normal subjects. TNF-alpha/IL-4, IL-2/IL-4, and IFN-gamma/IL-4 ratios were higher in preeclamptic patients than in the normal group. Significant positive correlations were observed between mean blood pressure and concentrations of the Th-1 type cytokines IL-2, IFN-gamma, and TNF-alpha. CONCLUSION: Up-regulation of Th1 responses and down-regulation of Th2 responses occur in preeclampsia. PMID- 10378025 TI - Immunoadsorption plasmapheresis as a treatment for pregnancy complicated by systemic lupus erythematosus with positive antiphospholipid antibodies. AB - PROBLEM: Our purpose was to study the effect of maternal immunoadsorption plasmapheresis (IA) on the outcome of pregnancies complicated by systemic lupus erythematosus (SLE) with positive antiphospholipid antibodies, which were known to have a strong correlation with abortion or stillbirth. METHOD OF STUDY: Eight pregnancies in 7 patients with SLE were treated according to our protocol. They were all positive for the lupus anticoagulant. The treatments provided in these cases were as follows: an oral low-dose steroid; oral low-dose aspirin; and IA. The outcomes of the pregnancies were then studied. RESULTS: Of eight pregnancies, seven resulted in preterm deliveries, and cesarean sections were performed at 26 36 weeks of gestation. In one case, intrauterine fetal death occurred at 24 weeks of gestation. The other seven pregnancies resulted in live births (survival rate of 87.5%). CONCLUSION: IA improves the outcome of pregnancy complicated by SLE with positive antiphospholipid antibodies, without increasing steroid dosage. PMID- 10378026 TI - Vertical transmission of HIV: parameters which might affect infection of placental trophoblasts by HIV-1: a review. Biomed Group on the Study of in Utero Transmission of HIV 1. AB - PROBLEM: To understand the mechanisms preventing and/or facilitating maternofetal transmission of human immunodeficiency virus (HIV)-1 across the placenta during pregnancy. METHODS OF STUDY: Current experimental data were reviewed. RESULTS AND CONCLUSIONS: The data about the production of cytokines by placental cells and explants, taken together with information indicating selective passage of certain HIV-1 variants across the placental trophoblast, suggest an intricate regulatory network operating at the fetomaternal interface. The data show a differential differentiation of early and late trophoblasts, as far as HIV entry routes are concerned. We believe this explains the relative predominance of the early infection window, as far as in utero infection is concerned. Whether such a differentiation state can be transiently induced on term placental trophoblasts by several differentiation agents, including cytokines, is being investigated. Whatever the results may be, it is obvious that infection of placental cells is an excellent model of passage infection by HIV of/through a mucosal barrier. PMID- 10378027 TI - Interleukin-12 augments cytolytic activity of peripheral and decidual lymphocytes against choriocarcinoma cell lines and primary culture human placental trophoblasts. AB - PROBLEM: Human trophoblasts are tolerant to the maternal immune system, but susceptible to interleukin (IL)-2-activated lymphocytes. IL-12 is also a key cytokine in the induction of cytotoxic responses. We administered IL-12 to peripheral blood lymphocytes (PBLs) and to decidual lymphocytes (DLs) and studied resulting cytotoxicity against trophoblasts. METHOD OF STUDY: PBLs and DLs were stimulated with rIL-2 and/or rIL-12 for 48 hr in vitro. Cytotoxicity against the choriocarcinoma cell line JEG-3, JAR, and primary culture trophoblasts were examined by LDH release assay. The proliferative response was estimated by MTT assay. Expression of cytotoxic factors was studied by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: Whereas IL-12 alone produced a modest enhancement in cytotoxicity of PBLs and DLs, the combination of IL-2 and IL-12 was most effective in trophoblast cell lysis. IL-12 enhanced the mRNA expression of T-cell specific serine protease (TSP, granzyme B) and FasL in DLs, but the expression of perforin was unchanged. Expression of these cytotoxic factors in PBLs was up-regulated by IL-12. CONCLUSION: Our findings indicate critical roles of IL-12 in the activation of maternal lymphocytes, which could possibly result in pregnancy failure syndromes. PMID- 10378028 TI - Quantification of C3 and C4 in infertile men with antisperm antibody in their seminal plasma. AB - PROBLEM: Previous studies showed that some infertilities are caused by antisperm antibodies (ASAs). It was shown that some major complement (C) components are present in seminal fluid. Due to the role of C in the pathogenesis of ASAs, the existence and amount of two key C components (C3 and C4) were investigated in seminal plasma (SP). METHOD OF STUDY: Single radial immunodiffusion assay and a sandwich-type enzyme-linked immunosorbent assay (ELISA) were used for C3 and/or C4 quantification, respectively, in serum and SP, and the tray agglutination test was used for ASA detection in 12 fertile and 53 infertile men (18 ASA-positive [ASA+] and 35 ASA-negative [ASA-] men). RESULTS: Of the 18 ASA + infertile men, 61.11% had positive C3, whereas 27.77% showed positive C4 levels. ASA + infertile men showed significant differences in seminal plasma C3 mean values compared with ASA- infertile (P < 0.01) and fertile (P < 0.05) men, but the seminal plasma C4 values only showed differences compared with ASA- infertile men (P < 0.05). No significant differences were observed in serum C3 and/or C4 levels of ASA+ infertile men compared with other groups. No significant correlation was found between ASA titer and C3 and C4 levels in SP. A significant correlation existed between SP and serum C3 levels of ASA+ (r = 0.522, P < 0.01) and ASA- (r = 0.451, P < 0.01) infertile men, but no correlation was observed between C4 levels. CONCLUSIONS: In the presence of ASAs, the C system has no definitive activity in blood serum or outside the male genital tract. In SP, and in association with ASAs, C has no lytic activity by the classical pathway. The excess of C3 in SP of ASA+ infertile men may participate in other C-mediated activities in the male reproductive tract. PMID- 10378029 TI - Decidual natural killer cells in recurrent spontaneous abortion with normal chromosomal content. AB - PROBLEM: The maternal local immune responses in unexplained recurrent spontaneous abortion (RSA) are not yet well known. Maternal peripheral and decidual natural killer (NK) cells were evaluated in RSA with normal chromosomal content. METHOD OF STUDY: Maternal peripheral blood, villous trophoblast, and decidua were taken from 15 normal pregnancies and 9 RSA patients with normal chromosomes. The NK cells in decidual lymphocytes were evaluated by flow cytometry using monoclonal antibodies for CD56, CD16, and CD3. RESULTS: The percentages of CD56+ CD16- CD3- cells in decidual lymphocytes in RSA were lower than in normal pregnancies (P < 0.002). The CD56+CD16+/CD56+CD16- cells ratio in RSA was higher than in normal pregnancies (P < 0.02). CONCLUSION: The lower percentages of CD56+CD16-CD3- cells in RSA cases may show an inappropriate accumulation of NK cells in the decidua, and this finding may be a factor involved in RSA. PMID- 10378030 TI - A possible role for activated complement component 3 in phagocytic activity exhibited by the mouse trophoblast. AB - PROBLEM: To determine whether any blood plasma factor may play a regulatory role in trophoblast phagocytosis in rodent early pregnancy. METHOD OF STUDY: The effects of alloplasma on the phagocytosis of cultured mouse trophoblast cells (TCs) were evaluated using erythrocytes as target cells, in the presence of 10% fresh, normal plasma; 10% heat-inactivated plasma; 10% component 3 (C3)-depleted plasma; or medium alone. The possible activation of C3 complement, the phagocytosis of zymosan bound or unbound to C3b, and immunoreactivity to C3b receptor were also estimated. Phagocytic activity was expressed as the percentage of phagocytic TCs, and as the number of phagosomes/TCs. RESULTS: The use of complement sufficient plasma significantly enhanced the phagocytosis of the TCs while the use of heat-inactivated plasma eliminated the erythrophagocytosis. Very low levels of phagocytic activity were seen when the plasma was C3-complement deficient. Phagocytosis of C3b-bound zymosan was remarkable in comparison to zymosan alone, and immunoreactivity to C3b-receptors was seen on the TCs. CONCLUSION: These results indicate the participation of thermosensitive molecules mediating the phagocytosis of TCs and suggest, as in macrophages, a role for C3 C3b in this process. PMID- 10378031 TI - Survival of fetuses and viruses: universal mechanisms of co-existence with an immunological potent host. PMID- 10378032 TI - Up-regulation of natural killer cell cytotoxicity by interleukin-2: the effect of sex and parity. AB - PROBLEM: Peripheral blood lymphocytes (PBLs) from some, but not all, female donors showed increased cytotoxicity in response to interleukin (IL)-2. METHOD OF STUDY: The effect of IL-2 on natural killer (NK) cell cytotoxicity was compared in nulliparous females, parous females, and males. Peripheral blood lymphocytes were preincubated for 20 or 72 hr with 5 or 100 U/ml IL-2 and cytotoxicity against K562 targets was then examined. RESULTS: In the parous females, only the 72-hr preincubation with 100 U/ml IL-2 significantly increased NK cell cytotoxicity, whereas nulliparous females also showed significantly increased cytotoxicity after a 20-hr preincubation with 100 U/ml IL-2. Neither female subject group had increased activity after preincubation for 20 or 72 hr with 5 U/ml IL-2. However, male peripheral blood lymphocytes also showed a significant increase in NK cell cytotoxicity when preincubated for 72 hr with 5 U/ml IL-2. CONCLUSIONS: The effect of IL-2 on NK cell cytotoxic activity may be related to sex and the state of parity. PMID- 10378033 TI - [ENT, fractals and chaos]. PMID- 10378034 TI - [Radiation treatment of early stage supraglottic cancer]. AB - The management of early-stage squamous cell carcinoma (SCC) of the supraglottic larynx is still controversial. Supraglottic laryngectomy as well as irradiation alone is correlated with good oncological and functional results. In order to evaluate the results and prognostic factors influencing the successful using radiotherapy (RT), the authors performed a retrospective study of 100 consecutive T1-T2 N0 M0 cases of SCC of the supraglottic larynx, treated at a single institution between 1983 and 1992. RT was delivered with 60Co or 6 MeV photons through two lateral parallel opposed portals encompassing the primary laryngeal tumor and the upper and mid-neck nodes (Robbins' levels II, III and V). Supraclavicular nodes (level IV) were electively irradiated in 54 patients with T2 N0 tumors only, using an anterior field with midline block. Sixty-three patients received conventional fractionation (2 Gy/fraction, once-a-day, five times a week), while 37 patients were irradiated according to a twice-a-day fractionation regimen (1.5 Gy/fraction, twice a day with six-hour interval, five days a week). The median total tumor dose delivered was 67 Gy. A multivariate analysis showed that performance status, tumor grade and fractionation modality were the only statistically significant variables influencing disease-free survival. Acute and late radiation reactions were relatively low. This retrospective study confirms that conservative management of T1-T2 N0 supraglottic cancer using RT can achieve good cure rates with the possibility of larynx preservation for the majority of the patients. The decision between different conservative treatment modalities may be influenced by several factors correlated to the patient's conditions, tumor characteristics, but especially treatment modalities. PMID- 10378035 TI - [Primary synovial sarcoma of head and neck. Materials of The Gustave Roussy Institute. Report of 13 cases]. AB - This report presents the result of our experience with 13 primary synovial sarcoma of the head and neck. The 9 males and 4 females had a median age of 32 years. The predominant location of the tumor was the pharynx, treatment consist of surgical excision alone or associated with post-operative radiotherapy or chemotherapy. Mean follow-up was 48 month. Local recurrence occurred in 2 patient, 4 patient died of pulmonary metastasis. The five year survival rate was 55%. Favorable prognostic findings included age < 20 years and complete initial resection. PMID- 10378036 TI - [Does allergic fungal sinusitis exist? Preliminary results of a prospective study]. AB - Allergic aspergillar sinusitis is a very controversial clinical feature. We present results of a prospective study aimed at evaluating the reality of allergic aspergillar sinusitis in a nosologic and clinical point of vue. During a 5 months period, 31 patients underwent surgery: 21 sino-nasal polyposis, 5 chronic sinusitis without polyposis, 5 chronic sinusitis with radiologic images evocative of mycosis. The study was carried out using clinical criteria (per operative discovery of glue-like, thickened and viscous aspect of the secretions), pathologic criteria (the presence of elements consitutive of allergic mucin), mycological criteria (direct examination and culture), and immunoallergic criteria (specific IgE for Aspergillus fumigatus, serology for Aspergillus fumigatus and Aspergillus flavus (IgM, IgG), skin tests for Aspergillus fumigatus). In three cases we suspect an allergic aspergillar sinusitis (one patient presenting a bilateral chronic sinusitis and two patients presenting a sinonasal polyposis). In two patients presenting a sinonasal polyposis, a allergic fungal sinusitis was suspected, fungal identification was not possible. PMID- 10378037 TI - [Acoustic neuroma in children. Report of 5 cases]. AB - In this report, we present five cases of acoustic neuroma in children; one of them concern a case of neurofibromatosis 2. The most common symptom is a deafness, but it can also be revealed by a tinnitus, a vertigo, a facial nerve paralysis or headache. Diagnosis is confirmed by T1 and T2-weighted MRI with intravenous infusion of gadolinium. The deals of the treatment are tumor's control and preservation of the hearing and facial nerve functions. The means used are either microsurgery with a preference for posteriors approaches to translabyrinthic one, or radiosurgery with gamma knife which seems to be a promising technique for stades II and III tumors. Only children with small acoustic neuroma and good hearing can be simply watched. In neurofibromatosis 2, acoustic neuromas are bilateral and can be associated with other neuromas and cutaneous or ophtalmologic symptoms. Surgery is indicated in first on the side of the biggest tumor or the worst hearing function. Only a few patients well selected may benefit from an auditory brainstem implant. PMID- 10378038 TI - [Hypopharyngeal sarcomatoid carcinoma with one time sarcomatous recurrence. A case report]. AB - We report case of a patient with hypopharyngeal biphasic sarcomatoid carcinoma, with two tumor cell component, epidermoid and spindle cells, treated by surgery only. Two years later, recurrence is noted with totally different histologic form, sarcomatous monophasic, with spindle tumor cells and smooth muscle phenotype with immunohistochemistry, looking like primitive leiomyosarcoma of high malignancy degree. Such monophasic sarcomatous forms of sarcomatoid carcinoma, without epithelial tumor component, are deceptive and can be mistaken with primitive sarcoma. That is why discovery of epithelial differenciation signs, with immunohistochemistry or electronic microscopy, is very important. In our case, while the epithelial differenciation in the recurrent tumor is away, previous history of true biphasic sarcomatoid carcinoma in the same anatomic location, allows to assert recurrence of the same tumor with monophasic sarcomatous expression and smooth muscle phenotype in this case. Therefore in front of "sarcoma of the upper aerodigestive tract", of any immunohistochemical phenotype, monophasic sarcomatous form of epidermoid sarcomatoid carcinoma have always to be evocated and searched. PMID- 10378039 TI - [Homicide by gunshot. An evaluation of 50 homicides with reference to gunshot wound site]. AB - In this paper the homicides committed in the region of Ulm with firearms over a period of 17 years were studied. These included 50 deaths caused by gunshot wounds, 33 males and 17 females. Homicides by firearms accounted for 44 percent, which was slightly higher than in other German regions. Homicide victims were found in all age groups with a markedly higher number up to the 20th year and in the fourth decade of life. In most cases small arms were used, and in many cases multiple shots were fired primarily hitting the head and the left side of the trunk: but injuries in the region of the extremities were also seen. PMID- 10378040 TI - [Fatal child abuse (caused by physical violence) in Germany during 1 January 1985 to 2 October 1990. Results of a multicenter study]. AB - No reliable data are available on cases of lethal child abuse (by active force) in the area of Federal Republic of Germany prior to reunification (the former West Germany). In a multicenter study we therefore examined the police and court records for such cases occurring in the period 1 January 1985 to 2 October 1990 in nearly the entire area of Federal Republic of Germany. RESULTS: The study center received information on 58 cases of lethal child abuse. Extrapolated to all institutes of legal medicine, this corresponds to 62 cases in all of West Germany in the period studied. An approximately equal number of unreported cases should be added to this figure. Including unreported cases, at least 20 cases of lethal child abuse occurred per year; thus only one in every two cases ever came to light. Almost two thirds of the victims were younger than one year old. At autopsy 59% exhibited signs of repeated abuse at autopsy. By far the most common cause of death was direct impact from a blunt object, usually to the head. Mostly, the male person to whom the victim relates most closely (father, stepfather, partner of the mother) has killed the child. Twenty-one of the 74 persons charged saw the charges against them dropped or were acquitted due to lack of evidence; 51 received sentences ranging from one year probation to life. In the remaining two cases the outcome of the trial was unknown. Signs of abuse were readily apparent at autopsy in almost all cases. The high number of unreported cases underscores the need to educate medical students and practicing physicians to be on the look-out for signs of abuse and argues for an increase in the rate of autopsy. PMID- 10378041 TI - [Forensic identification value of roentgen images in determining tooth-colored dental filling materials]. AB - Identification of unknown bodies is mainly made by dental examination and comparison with accurate dental records. Therefore it is necessary to examine the jaws carefully and to locate every tooth-coloured dental restoration. Overlooking dental fillings can make positive identification impossible. The paper presented here evaluates the usefulness of radiographs to locate tooth coloured dental restorations. RESULTS: 40% of the dental materials tested here could not be detected radiographically with a sensitivity that is demanded. A radiographic examination is not sufficient and must be supplied by other techniques. PMID- 10378042 TI - [Wound entry findings of animal anesthesia guns without smoke outlets]. AB - Subsequent to previous reports on the morphology of injuries from slaughterer's guns a case of suicide with atypical entrance wound findings is reported from the Freiburg autopsy material. The livestock stunner used did not have the smoke conduits usually opening into the muzzle end. Accordingly the entrance wound was not associated with any roundish or elliptic soot deposits. In cases where a large entrance hole without paired or cloverleaf-shaped zones of blackening, but no exit wound and no projectile is found one should also think of a slaughterer's gun without smoke conduits. The results of our test shots with conventional livestock narcotic devices and stunners without smoke conduits are presented. PMID- 10378043 TI - ["Crow's feet wrinkles" as a sign of preserved consciousness]. AB - We report on 3 deaths after the influence of high voltage current, fire and traumatic injury respectively. In all cases we found radial bands beside the eyes -so called "crow's feet". They are a sign of vital reaction in the cases of traumatic and thermic injury, possibly also in high-voltage burns due to sparking. In general, the "crow's feet" show that the person was conscious at the moment of the event. PMID- 10378045 TI - [Airway remodelling in asthma]. PMID- 10378044 TI - [Differential diagnostic aspects of forensically relevant findings in the brain stem]. AB - Because of the extreme dense accumulation of vital structures (compared with other regions of the central nervous system), in the brainstem even small lesions may cause serious clinical symptoms. Judging the forensic relevance of macroscopically visible lesions requires the knowledge of the respective possible diagnosis. As shown in three case reports (67 years, teleangiectasis; 35 years, cavernoma; 49 years, secondary hemorrhage following trauma) this demands apart from the knowledge of the normal and pathological anatomy of the brainstem always the use of histological methods. PMID- 10378046 TI - [Evaluating the influence of hospital size on respiratory health care efficiency in Andalusia]. AB - OBJECTIVE: To analyze the influence of hospital size on efficient delivery of health care to patients admitted with respiratory problems in Andalusia (Spain). METHOD: From the minimal data base of all public hospital releases from Andalusia for 1994 and 1995, we identified patients whose main diagnosis involved respiratory complaints as shown by diagnosis-related grouper AP-DRG-10.0, excluding patients aged under 15 years. A functional index for each hospital was calculated to estimate hospital efficiency; a case-mix index was used to express the complexity of cases attended. RESULTS: 1) The case-mix indexes of the hospitals studied were not significantly different. Functional indexes were positively related to hospital size. CONCLUSIONS: 1) In Andalusia, case complexity does not increase with the size of public hospital. 2) Larger hospital size led to decreased efficiency in caring for respiratory patients in the population and period under study. PMID- 10378047 TI - [Efficacy of demand of positive airway pressure therapy for treating obstructive sleep apnea syndrome]. AB - The treatment of choice for obstructive sleep apnea syndrome (OSAS) is nasal continuous positive airway pressure (nCPAP). The precise level of pressure is adjusted by polysomnography. Devices to deliver pressure on demand have recently been designed to adapt the level of pressure to each respiratory cycle according to flow modification. OBJECTIVE: To compare the manual titering nCPAP system with that of demand continuous positive airway pressure (nDPAP) in patients diagnosed of OSAS. PATIENTS AND METHODS: Eighteen consecutive patients whose OSAS was diagnosed by conventional polysomnography were enrolled with apnea-hypopnea indexes over 10/hour (AHI > 10) and clinical symptoms of daytime drowsiness and/or cardiovascular risk factors. Titering polysomnographs were performed for all patients with nCPAP and with nDPAP and analyzed blindly. RESULTS: No significant differences between nCPAP and nDPAP were found in neurophysiological variables analyzed (sleep architecture, arousals, sleep efficiency) or in respiratory variables (AHI, oxygen saturation) with the exception of minimum SatO2 during REM sleep, which was significantly better with nCPAP (p < 0.03). Mean end pressure with nCPAP and mean pressure with nDPAP were similar; it is also worth noting that mean pressure was lower with nDPAP than with titered nCPAP pressure a mean 65.7 +/- 22% of the time. CONCLUSION: Automatic nDPAP is as effective as titered nCPAP for treating patients with OSAS. PMID- 10378048 TI - [Video-assisted thoracoscopy for resecting solitary pulmonary nodules]. AB - The objective of this study was to evaluate the usefulness of video assisted thoracoscopy in the resection of solitary pulmonary nodules. Thirty-three patients with solitary pulmonary nodules diagnosed by video assisted thoracoscopy were enrolled prospectively. A preoperative computed tomography scan was obtained for each patient. Harpoons were implanted preoperatively to locate the lesion in patients whose tumors were in the parenchyma. When endoscopic resection proved impossible in five patients, the surgeon resorted to thoracotomy. All were diagnosed after the procedure. One was a case of pulmonary lymphoma, 2 were primary adenocarcinomas of the lung, 2 were oat-cell cancers, 1 was Wegener's disease, 4 were tuberculomas, 3 involved pulmonary infarction and 20 were single pulmonary metastases. Patients who needed thoracotomy required more days of postsurgical drainage (p < 0.05). The size of resected nodules ranged from 0.4 to 6 centimeters. Preoperative positron emission tomographs were available for four patients. No perioperative (< 30 days) mortality occurred and morbidity consisted of one case of prolonged airway leak (> 7 days). Use of video-assisted thoracoscopy reduced perioperative morbidity and hospital stay. PMID- 10378049 TI - [Do patients lie about smoking during follow-up in the respiratory medicine clinic?]. AB - Quitting smoking is a first-line treatment for patients with bronchial diseases. Continued smoking worsens the clinical course of chronic broncho-pulmonary diseases and increases the number of exacerbations. Specialists commonly insist on the need to quit smoking. This study sought to determine whether a percentage of patients seen in a respiratory medicine clinic continued to smoke while denying doing so. One hundred twenty-five subjects were studied consecutively. At a regular visit they were first asked about smoking; later, without prior warning, exhaled carbon monoxide (CO) was measured by co-oximetry. If CO was over 10 ppm, the subject was considered to have been smoking. We defined a patient as a "liar" if he or she denied smoking but had a reading of CO in exhaled air over 10. Of the 125 cases studied, 21 (17%) smoked while denying doing so. Among men the percentage was 21%, and among ex-smokers, the figure was 27%. The highest value, 34%, was found among patients with chronic obstructive pulmonary disease (COPD). We conclude, therefore, that a substantial proportion of patients lies to their physicians. A third of COPD patients, who are particularly sensitive to the toxic effects of smoking, try to mislead their doctors. PMID- 10378050 TI - [European Asthma Study. Identifying and treating young adults with epidemiological criteria for asthma in five areas of Spain. Spanish Group of the European Asthma Study]. AB - BACKGROUND AND OBJECTIVES: Asthma's great impact on public health stems from its chronicity and to high prevalence among all age groups and both sexes. To estimate the appropriateness of treatment and management of asthma in Spain during the period of 1991 and 1992, we analyzed data from the European Community Respiratory Health Survey (ECRHS). METHOD: The ECRHS was undertaken with a random sample of 20-to-44-year-olds in Albacete, Barcelona, Galdakao, Huelva and Oviedo. In total, 181 individuals with asthma were identified. Current asthma was defined as the presence of respiratory symptoms associated with asthma within the past 12 months and a positive methacholine challenge test. RESULTS: Subjects who were unaware of having asthma made up 57.5% (CI: 49.9-64.8%), and 35.9% (CI: 27.9 42.3%) were not following any specific treatment. Among asthmatics who reported having continuous or frequent respiratory symptoms, 25.9% (CI: 15.3-39.0%) were not following any treatment. CONCLUSION: Over half the individuals with asthma in 1991 to 1993 were unaware of having the disease at the time of the study or of ever having had it, and approximately one third were not in treatment. The delivery of appropriate treatment in asthma generally, and in asthma with continuous or frequent respiratory symptoms, was markedly inadequate during the period studied. PMID- 10378051 TI - [Reduction in tobacco consumption. A strategy toward quitting smoking?]. PMID- 10378052 TI - [New frontiers in imaging diagnosis in pneumology]. PMID- 10378053 TI - [Kartagener's syndrome. Diagnosis in a 75 year-old woman]. AB - Kartagener's syndrome is an inherited disease characterized by the clinical triad of bronchiectasis, sinusitis and situs inversus caused by an ultrastructural defect in the cilia that results in impaired mucociliary clearance. It is usually diagnosed during childhood, with a small number of cases discovered in adults and even fewer among patients over 60 years of age. Prompt, appropriate treatment of respiratory infections can minimize irreversible lung damage. We report the case of a woman with bronchiectasis, sinusitis and situs inversus diagnosed of Kartagener's syndrome based on clinical signs at age 75 years. PMID- 10378054 TI - [Synchronous multiple primary cancer of the lung: a rare association of small cell carcinoma as the main tumor plus epidermoid carcinoma]. AB - Multiple primary cancer (MPC), a rare finding, is most often seen in the breast. In the lung, this cancer is rare (accounting for between 1.5 and 3% of cases), with epidermoid carcinoma usually being the principal tumor. The presentation of small-cell carcinoma as the principal tumor in MPC is thought to be extremely rare. The criteria for pulmonary MPC described by Martini and Melamed continue to provide the definition of reference. Pulmonary MPC can manifest in a synchronically (simultaneous appearance) or metachronically (with more than two years' lapse in appearance). Patients so-affected are usually male and heavy smokers. Survival with this type of lung cancer is usually less than for a single form. We report the case of a patient with synchronous MPC of the lung, with small cell carcinoma as the principal tumor associated with epidermoid carcinoma. We review the literature on this subject. PMID- 10378055 TI - [On the difference between classification and diagnostic criteria]. PMID- 10378057 TI - [Diuretics to treat asthma?]. PMID- 10378058 TI - [Pleural effusion secondary to intrathoracic migration of the distal end of a ventriculoperitoneal shunt]. PMID- 10378059 TI - [Tuberculous pseudo-chylothorax with normal deaminase adenosine]. PMID- 10378060 TI - Science or show business. PMID- 10378061 TI - "Living high and training low" can improve sea level performance in endurance athletes. PMID- 10378062 TI - Cricket: injury in long trousers. PMID- 10378063 TI - Seeking misclassification: "doping" in disability sport. PMID- 10378064 TI - Boxing and medicine. PMID- 10378065 TI - Musculoskeletal change during spaceflight: a new view of an old problem. PMID- 10378066 TI - Abdominal injuries and sport. AB - Serious abdominal injuries resulting from sport are rare. The potential for misdiagnosis is significant and the consequences may be serious. Patients with abdominal pain should be taken very seriously and investigated with appropriate diagnostic equipment. Sporting bodies have a responsibility to address safety within a particular sport and to change the rules where necessary as injury patterns are identified. PMID- 10378068 TI - How I treat: return to sport after post-viral fatigue. PMID- 10378067 TI - Diabetes and exercise. AB - Exercise is frequently recommended in the management of type 1 and 2 diabetes mellitus and can improve glucose uptake by increasing insulin sensitivity and lowering body adiposity. Both alone and when combined with diet and drug therapy, physical activity can result in improvements in glycaemic control in type 2 diabetes. In addition, exercise can also help to prevent the onset of type 2 diabetes, in particular in those at higher risk, and has an important role in reducing the significant worldwide burden of this type of diabetes. Recent studies have improved our understanding of the acute and long term physiological benefits of physical activity, although the precise duration, intensity, and type of exercise have yet to be fully elucidated. However, in type 1 diabetes, the expected improvements in glycaemic control with exercise have not been clearly established. Instead significant physical and psychological benefits of exercise can be achieved while careful education, screening, and planning allow the metabolic, microvascular, and macrovascular risks to be predicted and diminished. PMID- 10378069 TI - Injury rates in Shotokan karate. AB - OBJECTIVE: To document the injury rate in three British Shotokan karate championships in consecutive years. In these tournaments strict rules governed contact, with only "light" or "touch" contact allowed. Protective padding for the head, hands, or feet was prohibited. METHODS: Prospective recording of injuries resulting from 1770 bouts in three national competitions of 1996, 1997, and 1998. Details of ages and years of karate experience were also obtained. RESULTS: 160 injuries were sustained in 1770 bouts. The overall rate of injury was 0.09 per bout and 0.13 per competitor. 91 (57%) injuries were to the head. The average age of those injured was 22 years, with an average of nine years of experience in karate. CONCLUSIONS: The absence of protective padding does not result in higher injury rates than in most other series of Shotokan karate injuries. Strict refereeing is essential, however, to maintain control and minimise contact. PMID- 10378070 TI - Analysis of the aerobic-anaerobic transition in elite cyclists during incremental exercise with the use of electromyography. AB - OBJECTIVES: To investigate the validity and reliability of surface electromyography (EMG) as a new non-invasive determinant of the metabolic response to incremental exercise in elite cyclists. The relation between EMG activity and other more conventional methods for analysing the aerobic-anaerobic transition such as blood lactate measurements (lactate threshold (LT) and onset of blood lactate accumulation (OBLA)) and ventilatory parameters (ventilatory thresholds 1 and 2 (VT1 and VT2)) was studied. METHODS: Twenty eight elite road cyclists (age 24 (4) years; VO2MAX 69.9 (6.4) ml/kg/min; values mean (SD)) were selected as subjects. Each of them performed a ramp protocol (starting at 0 W, with increases of 5 W every 12 seconds) on a cycle ergometer (validity study). In addition, 15 of them performed the same test twice (reliability study). During the tests, data on gas exchange and blood lactate levels were collected to determine VT1, VT2, LT, and OBLA. The root mean squares of EMG signals (rms-EMG) were recorded from both the vastus lateralis and the rectus femoris at each intensity using surface electrodes. RESULTS: A two threshold response was detected in the rms-EMG recordings from both muscles in 90% of subjects, with two breakpoints, EMGT1 and EMGT2, at around 60-70% and 80-90% of VO2MAX respectively. The results of the reliability study showed no significant differences (p > 0.05) between mean values of EMGT1 and EMGT2 obtained in both tests. Furthermore, no significant differences (p > 0.05) existed between mean values of EMGT1, in the vastus lateralis and rectus femoris, and VT1 and LT (62.8 (14.5) and 69.0 (6.2) and 64.6 (6.4) and 68.7 (8.2)% of VO2MAX respectively), or between mean values of EMGT2, in the vastus lateralis and rectus femoris, and VT2 and OBLA (86.9 (9.0) and 88.0 (6.2) and 84.6 (6.5) and 87.7 (6.4)% of VO2MAX respectively). CONCLUSION: rms-EMG may be a useful complementary non-invasive method for analysing the aerobic-anaerobic transition (ventilatory and lactate thresholds) in elite cyclists. PMID- 10378071 TI - A preliminary study of patient comfort associated with customised mouthguards. AB - OBJECTIVE: To compare patient perception of custom made mouthguards of ideal and less than ideal designs in terms of their comfort and "wearability". METHOD: A mouthguard of ideal design (A) and one incorporating common design faults of underextension and unadjusted occlusion (B) were provided for 22 active sportsmen and women. They were not informed of the details of the design or the status of the protector. Half the participants were asked to wear mouthguard A first and the other half wore B first, each worn for one hour on two consecutive nights. Questionnaires were used to evaluate and rate the comfort and wearability of each mouthguard. RESULTS: Eighteen people completed the study. The ideal appliance was rated as significantly more retentive and comfortable overall and specifically was more comfortable to lips, gums, and tongue. It was also recognised as being less bulky, less likely to keep the teeth apart, or to cause pain in the jaw muscles. CONCLUSIONS: Comfort is likely to be increased if mouthguards are extended labially to within 2 mm of the vestibular reflection, adjusted to allow even occlusal contact, rounded at the buccal peripheries, and tapered at the palatal edges. PMID- 10378072 TI - Effect of 14 weeks of resistance training on lipid profile and body fat percentage in premenopausal women. AB - OBJECTIVES: To study the effects of a supervised, intensive (85% of one repetition maximum (1-RM)) 14 week resistance training programme on lipid profile and body fat percentage in healthy, sedentary, premenopausal women. SUBJECTS: Twenty four women (mean (SD) age 27 (7) years) took part in the study. Subjects were randomly assigned to either a non-exercising control group or a resistance exercise training group. The resistance exercise training group took part in supervised 45-50 minute resistance training sessions (85% of 1-RM), three days a week on non-consecutive days for 14 weeks. The control group did not take part in any structured physical activity. RESULTS: Two way analysis of variance with repeated measures showed significant (p < 0.05) increases in strength (1-RM) in the exercising group. There were significant (p < 0.05) decreases in total cholesterol (mean (SE) 4.68 (0.31) v 4.26 (0.23) mmol/1 (180 (12) v 164 (9) mg/dl)), low density lipoprotein (LDL) cholesterol (2.99 (0.29) v 2.57 (0.21) mmol/l (115 (11) v 99 (8) mg/dl), the total to high density lipoprotein (HDL) cholesterol ratio (4.2 (0.42) v 3.6 (0.42)), and body fat percentage (27.9 (2.09) v 26.5 (2.15)), as well as a strong trend towards a significant decrease in the LDL to HDL cholesterol ratio (p = 0.057) in the resistance exercise training group compared with their baseline values. No differences were seen in triglycerides and HDL cholesterol. No changes were found in any of the measured variables in the control group. CONCLUSIONS: These findings suggest that resistance training has a favourable effect on lipid profile and body fat percentage in healthy, sedentary, premenopausal women. PMID- 10378073 TI - A prospective epidemiological study of injuries in four English professional football clubs. AB - OBJECTIVE: To define the causes of injuries to players in English professional football during competition and training. METHOD: Lost time injuries to professional and youth players were prospectively recorded by physiotherapists at four English League clubs over the period 1994 to 1997. Data recorded included information related to the injury, date and place of occurrence, type of activity, and extrinsic Playing factors. RESULTS: In all, 67% of all injuries occurred during competition. The overall injury frequency rate (IFR) was 8.5 injuries/1000 hours, with the IFR during competitions (27.7) being significantly (p < 0.01) higher than that during training (3.5). The IFRs for youth players were found to increase over the second half of the season, whereas they decreased for professional players. There were no significant differences in IFRs for professional and youth players during training. There were significantly (p < 0.01) injuries in competition in the 15 minute periods at the end of each half. Strains (41%), sprains (20%), and contusions (20%) represented the major types of injury. The thigh (23%), the ankle (17%), knee (14%), and lower leg (13%) represented the major locations of injury, with significantly (p < 0.01) more injuries to the dominant body side. Reinjury counted for 22% of all injuries. Only 12% of all injuries were caused by a breach of the rules of football, although player to player contact was involved in 41% of all injuries. CONCLUSIONS: The overall level of injury to professional footballers has been showed to be around 1000 times higher times higher than for industrial occupations generally regarded as high risk. The high level of muscle strains, in particular, indicates possible weakness in fitness training programmes and use of warming up and cooling down procedures by clubs and the need for benchmarking players' levels of fitness and performance. Increasing levels of injury to youth players as a season progresses emphasizes the importance of controlling the exposure of young players to high levels of competition. PMID- 10378074 TI - Trace elements and electrolytes in human resting mixed saliva after exercise. AB - OBJECTIVES: Exercise is known to cause changes in the concentration of salivary components such as amylase, Na, and Cl. The aim of this investigation was to evaluate the effect of physical exercise on the levels of trace elements and electrolytes in whole (mixed) saliva. METHODS: Forty subjects performed a maximal exercise test on a cycle ergometer. Samples of saliva were obtained before and immediately after the exercise test. Sample concentrations of Fe, Mg, Sc, Cr, Mn, Co, Cu, Zn, Se, Sr, Ag, Sb, Cs, and Hg were determined by inductively coupled plasma mass spectrometry and concentrations of Ca and Na by atomic absorption spectrometry. RESULTS: After exercise, Mg and Na levels showed a significant increase (p < 0.05) while Mn levels fell (p < 0.05). Zn/Cu molar ratios were unaffected by exercise. CONCLUSIONS: Intense physical exercise induced changes in the concentrations of only three (Na, Mg, and Mn) of the 16 elements analysed in the saliva samples. Further research is needed to assess the clinical implications of these findings. PMID- 10378075 TI - Role conflict and confidentiality in multidisciplinary athlete support programmes. AB - As medical and scientific staff have increasingly been called upon to provide multidisciplinary support to elite performers the potential for ethical, professional, and legal conflicts has also increased. Although this has been recognised, little guidance has been provided to help resolve such conflicts. This paper identifies key issues in the provision of effective support and specifically addresses the roles of medical and scientific staff and their relations to coaches and performers. An athlete charter is presented that has successfully been used to resolve ethical conflicts and clarify the lines of communication, confidentiality, and responsibility within a national governing body. PMID- 10378076 TI - Injuries to riders in the cross country phase of eventing: the importance of protective equipment. AB - OBJECTIVES: To determine the distribution of injuries in the eventing discipline of equestrian sports and the effectiveness of the protective equipment worn. METHODS: Data on all injuries sustained in the cross country phase over fixed obstacles were collected from 54 days of competition from 1992 to 1997. This involved 16,940 rides. RESULTS: Data on a total of 193 injuries were collected, which included two deaths. This represents an injury rate of 1.1%. Head and facial injuries represented the largest group (31%), with one third of these requiring treatment in hospital. All riders were wearing protective helmets and body protectors. CONCLUSIONS: Eventing is one of the most dangerous equestrian sports. Improved protective equipment, which is mandatory for 1999, should reduce the severity of these injuries. PMID- 10378077 TI - The prevalence of chronic knee injury in triathletes. AB - OBJECTIVES: To add to the area of triathlon research by providing much needed prevalence data on knee injury in triathletes. METHOD: An incidental "in field" sampling technique was used to interview 58 triathletes aged between 15 and 55 years about knee injury during a triathlon event. The sample comprised 46 men and 12 women. RESULTS: Most knee injuries occurred during the running event (72%) and affected the lateral side of the knee (38%). In all, 78% of the sample sought treatment from a healthcare professional. CONCLUSION: The study has provided much needed prevalence data on chronic knee injury in triathletes. PMID- 10378078 TI - Drug testing. PMID- 10378079 TI - Calcium fluxes in hydrozoan embryos depend, in part, on exocytosis and fluid phase endocytosis. AB - In the hydrozoan Phialidium gregarium, the constitutive calcium influx of cleavage stage embryos in sea water is 1.96 +/- 0.75 x 10(-15) moles/embryo/minute. Treating embryos with 227 mM KCl in seawater briefly increases the calcium influx more than 100-fold, to 3.9 x 10(-13) mol/embryo/min. About 62% of the KCl-induced calcium influx is due to calcium flowing through voltage-sensitive calcium channels. This causes a marked intracellular calcium transient and secretion of intracellular vesicles. The other component (approximately 38%) of the calcium influx occurs via fluid phase endocytosis of the extracellular medium (detected using extracellular 3H-sucrose). KCl-treatment of 45Ca loaded embryos induces a 45Ca efflux which can reach peak fractional rates of 0.98/min, during which 55-75% (mean 66%) of the total 45Ca is lost. The KCl-induced calcium efflux is due, in part, to secretion because loaded 3H sucrose is effluxed simultaneously. This pathway may be important for the calcium efflux necessary for long-term calcium homeostasis in cells. PMID- 10378080 TI - Modulation of Ca2+ channel-gated Ca2+ release by W-7 in cardiac myocytes. AB - Cardiac muscle excitation-contraction coupling is controlled by the Ca(2+) induced Ca2+ release mechanism. The present study examines the effects of a calmodulin antagonist W-7 on Ca2+ current (ICa)-induced Ca2+ release in whole cell-clamped rat ventricular myocytes. Exposure of cells to W-7 suppressed ICa, but the intracellular Ca(2+)-transients showed a lesser degree of reduction, suggesting possible enhancement of Ca(2+)-induced Ca2+ release. The effects of W 7 on the efficacy of Ca2+ release were most prominent at negative potentials. At test potentials of -30 mV, 20 microM W-7 almost completely blocked ICa, but significant Ca(2+)-transients remained, thus causing a four to six-fold increase in the efficacy of Ca(2+)-induced Ca2+ release. The depolarization-dependent Ca(2+)-transients were eliminated in absence of extracellular Ca2+, blocked by Cd2+, and were absent when the sarcoplasmic reticulum was depleted of Ca2+, implicating dependency on Ca(2+)-signaling between the L-type channel and the ryanodine receptor. W-7 mediated increase in the efficacy of Ca(2+)-induced Ca2+ release was eliminated when myocytes were dialyzed with the internal solution containing gluathione (5 mM), suggesting the possible role of cellular redox state in the regulation of Ca2+ release by the calmodulin antagonist. PMID- 10378081 TI - Visualization of biphasic Ca2+ diffusion from cytosol to nucleus in contracting adult rat cardiac myocytes with an ultra-fast confocal imaging system. AB - In contracting cardiac myocytes, the rapid changes in cytosolic and nuclear Ca2+ make it difficult to determine whether the nuclear Ca2+ transient is caused by diffusion from the cytosol or by Ca2+ release channels on the inner nuclear membrane, or both. The propagation mechanism in the nucleoplasm also remains unknown. We have developed an ultra-fast Nipkow confocal imaging system able to acquire two-dimensional images at approximately 4 ms/full frame speed and employed it to analyze Ca2+ waves and the dynamics of the cytosolic and nuclear Ca2+ transients after electrical stimulation of cardiac myocytes. The pattern of nuclear Ca2+ upon stimulation was well described by a mathematical model of Ca2+ diffusion across the nuclear envelope. No evidence of Ca2+ release from perinuclear Ca2+ stores was obtained. The Ca2+ diffusion constant appeared to change during contraction, with essentially free diffusion of Ca2+ through nuclear pore complexes at low cytosolic Ca2+ and partially restricted diffusion at high cytosolic Ca2+. The Ca2+ in the nucleoplasm propagated by diffusion and no Ca2+ release phenomena were seen in the nucleus. PMID- 10378082 TI - Different contributions of voltage-sensitive Ca2+ channels to histamine-induced catecholamine release and tyrosine hydroxylase activation in bovine adrenal chromaffin cells. AB - Histamine stimulates catecholamine release and tyrosine hydroxylase activity in a Ca(2+)-dependent manner in bovine adrenal chromaffin cells. The role of voltage sensitive Ca2+ channels in these two responses has been investigated. Using an EC50 concentration of histamine, 1 microM, catecholamine release was enhanced by (+/-)BayK8644, and partially inhibited by nitrendipine and omega-agatoxin IVA, blockers of L- and P/Q-type Ca2+ channels. omega-Conotoxin GVIA gave small and variable inhibitory effects. With a maximal histamine concentration, 10 microM, similar results were obtained except that now omega-conotoxin GVIA reliably reduced release. In contrast, neither (+/-)BayK8644 nor any of the individual Ca2+ channel antagonists had any significant effect on tyrosine hydroxylase (TOH) activation induced by either an EC50 or a maximal concentration of histamine. When high concentrations of nitrendipine, omega-conotoxin GVIA and omega-agatoxin IVA were combined with omega-conotoxin MVIIC (a non-selective blocker of N, P and Q channels) to block voltage-sensitive Ca2+ channels in these cells, release induced by K+ depolarization was completely blocked. Release caused by histamine, however, was substantially reduced but not abolished. The combination of antagonists also only partially inhibited TOH activation by histamine. The results show that the G protein-coupled receptor agonist histamine activates several different types of voltage-sensitive Ca2+ channels in chromaffin cells to mediate its cellular effects. Histamine may also activate additional pathways for Ca2+ entry. The results also suggest that the manner by which Ca2+ controls release and TOH activation once it has entered chromaffin cells through these channels are different. PMID- 10378083 TI - Regulation of Ca2+ release by cAMP-dependent protein kinase. A mechanism for agonist-specific calcium signaling? AB - Calcium is an ubiquitous second messenger that is involved in the regulation of a number of cell functions. The mechanism by which the specificity of calcium signaling is achieved is not well understood. We suggest that calcium release from the ER can occur selectively at different spatial locations in response to different extracellular stimuli. We discuss a possible mechanism for such selectivity and present a model based on this mechanism. The suggested mechanism is based on the regulation of local Ca2+ release by cyclic AMP-dependent protein kinase (PKA) and relies upon two experimental observations: first, some G-protein coupled signaling pathways activate PLC and regulate adenylate cyclase at the same time, leading to IP3 production and altering PKA activity via changes in cAMP level; second, phosphorylation by PKA alters the properties of IP3 receptor (IP3R). In our model we consider allosteric regulation of IP3Rs by IP3 and cAMP dependent phosphorylation. The differences in IP3Rs and PKA densities at different spatial locations within the cell allow the release of calcium selectively at each location in response to certain combination of IP3 and cAMP concentration. Specificity of agonist-response coupling is achieved if different combinations in the levels of these second messengers are specific for different extracellular stimuli. PMID- 10378084 TI - Measurement of sarcoplasmic reticulum Ca2+ content in intact amphibian skeletal muscle fibres with 4-chloro-m-cresol. AB - Single skeletal muscle fibres were isolated from the toad (Bufo marinus) and isometric force and myoplasmic free calcium concentration ([Ca2+]i) were measured. Brief applications of 4-chloro- m-cresol (4-CmC, 0.2-5 mM) elevated [Ca2+]i reversibly in a dose-dependent manner. The lowest concentration of 4-CmC which reliably gave maximal [Ca2+]i was 2 mM and it was, therefore, used for measurement of sarcoplasmic reticulum (SR) Ca2+ content. Tetanic stimulations (100 Hz) increased [Ca2+]i from a resting level of 105 +/- 47 nM (n = 10) to 1370 +/- 220 nM (n = 6). Application of 2 mM 4-CmC produced a contracture that was 54 +/- 16% (n = 6) of the tetanic force and elevated [Ca2+]i to a peak of 3520 +/- 540 nM (n = 8). Both force and [Ca2+]i levels (resting and tetanic) were restored after 10 min of washout of 4-CmC. In skinned muscle fibres, the myofibrillar Ca(2+)-sensitivity was not changed by 4-CmC, but maximal force was reduced to 74 +/- 10% (n = 4). The magnitude of the peak of the 4-CmC-induced Ca2+ transient was not significantly changed by removal of extracellular Ca2+ nor by inhibiting the SR Ca2+ pump with 2,5-di-tert-butylhydroquinone. Treatment of intact fibres with 30 mM caffeine produced a peak Ca2+ level that was indistinguishable from 2 mM 4-CmC. These results indicate that it is possible to measure the SR Ca2+ content in the same fibre with 4-CmC without loss of normal muscle function. PMID- 10378085 TI - What drives calcium entry during [Ca2+]i oscillations?--challenging the capacitative model. AB - An increased entry of Ca2+ across the plasma membrane plays a key role in the generation and maintenance of the [Ca2+]i signals seen in cells following activation of receptors coupled to the PLC/InsP3 signaling pathway. In recent years, considerable efforts have been made to define the nature and control of this agonist-enhanced Ca2+ entry. To date, these studies have largely focussed on the so-called 'capacitative' or store-operated model and, although many important details remain unclear, the critical role this mechanism plays in maintaining the sustained elevated 'plateau' type of [Ca2+]i response seen at high agonist concentrations is now well established. Far less well understood is the nature of the enhanced Ca2+ entry associated with the more complex [Ca2+]i signals typical of stimulation at more physiological levels of agonist. Where such entry has been considered, it too has generally been assumed to result from a capacitative or 'store-operated' mechanism. Significantly, however, direct evidence in support of this assumption is lacking. This review attempts to critically examine this assumption and presents the argument that several key characteristics of capacitative or store-operated mechanisms of agonist-activated Ca2+ entry are incompatible with its operation during these types of [Ca2+]i signal. PMID- 10378086 TI - Molecular properties of inositol 1,4,5-trisphosphate receptors. AB - The receptors for the second messenger inositol 1,4,5-trisphosphate (IP3) constitute a family of Ca2+ channels responsible for the mobilization of intracellular Ca2+ stores. Three different gene products (types I-III) have been isolated, encoding polypeptides which assemble as large tetrameric structures. Recent molecular studies have advanced our knowledge about the structure, regulation and function of IP3 receptors. For example, several Ca(2+)-binding sites and a Ca(2+)-calmodulin-binding domain have been mapped within the type I IP3 receptor, and studies on purified cerebellar IP3 receptors propose a second Ca(2+)-independent calmodulin-binding domain. In addition, minimal requirements for the binding of immunophilins and the formation of tetramers have been identified. Overexpression of IP3 receptors has provided further clues to the regulation of individual IP3 receptor isoforms present within cells, and the role that they play in the generation of IP3-dependent Ca2+ signals. Inhibition of IP3 receptor function and expression, and analysis of mutant IP3 receptors, suggests that IP3 receptors are involved in such diverse cellular processes as proliferation and apoptosis and are thus, necessary for normal development. Our understanding of the complex spatial and temporal nature of cytosolic Ca2+ increases and the role that these Ca2+ signals play in cell function depend upon our knowledge of the structure and the regulation of IP3 receptors. This review focuses on the molecular properties of these ubiquitous intracellular Ca2+ channels. PMID- 10378087 TI - Slow changes in cytosolic free Ca2+ in Escherichia coli highlight two putative influx mechanisms in response to changes in extracellular calcium. AB - Free intracellular Ca2+ ([Ca2+]i) in Escherichia coli was measured using the bioluminescent protein aequorin. Overall, the bacteria maintained a tight control on their free [Ca2+]i. The results indicated a slow Ca2+ influx, the magnitude of the initial rise in free [Ca2+]i being dependent upon the concentrations of external Ca2+. This was followed by the slow removal of free Ca2+ until normal levels were restored. Specifically, addition of external Ca2+ (0.25-10 mM) resulted in a gradual rise in intracellular free Ca2+ from a basal level of approximately 272 nM, maximally reaching a peak of 0.85-5.4 microM within 30-40 min. This was followed by a slow fall over the next 30 min, culminating in an oscillatory pattern of free [Ca2+]i (range 0.3-0.7 microM for 0.25 mM external Ca2+). In the presence of EGTA, free [Ca2+]i was dramatically reduced. Neither the influx of Ca2+ nor restoration of intracellular free Ca2+ required protein synthesis. Moreover, preincubation with Ca2+ increased the rising phase of intracellular Ca2+ in response to further exposure to external Ca2+. This was further evidence against a specific adaptation process such as the synthesis of calcium exporters. A putative Ca2+ influx channel was demonstrated in stationary phase cells in particular, which could be blocked by La3+. This channel was consistent with the voltage-activated poly-3-hydroxybutyrate/polyphosphate Ca2+ channels previously detailed by Reusch et al. [23] Even in the presence of La3+, however, the free [Ca2+]i of log phase and stationary phase bacteria still increased two-fold over resting values in response to external Ca2+. This suggested the presence of at least two Ca2+ influx processes, one inhibited by La3+ and the other not. PMID- 10378088 TI - A critical review of cranial ultrasounds: is there a closer association between intraventricular blood, white matter abnormalities or cysts, and cerebral palsy? AB - In an attempt to determine cranial ultrasonographic features of preterm infants with intraventricular hemorrhage (IVH) and/or periventricular white matter abnormalities (PVWMA) that could serve as more specific predictors of cerebral palsy (CP), we reviewed the cranial sonograms of 34 infants with IVH and/or PVWMA. Fourteen of the 34 infants studied (41%) developed CP. One of five infants with grade III IVH alone developed CP. Eleven infants with PVWMA did not develop cysts and only two (18%) developed CP (p = 0.04). Of the 18 infants who went on to develop cysts, four had a small, discrete solitary cyst and 14 had large cystic areas. Three of the four with small cysts were neurologically normal, whereas only three of the 14 with large cysts were neurologically normal (p = 0.04). Preterm infants with grade III IVH in the absence of any parenchymal lesion had a more favorable neurologic outcome than those with IVH and concomitant PVWMA. Infants with PVWMA in the presence or absence of IVH had much poorer neurologic prognoses. In infants with PVWM abnormalities, both the presence and extent of cystic lesions, though not their location, are the strongest predictors of long-term neurologic outcome. PMID- 10378089 TI - Children with moderately elevated lead levels: is chelation with DMSA helpful? AB - This study evaluates the effectiveness (use under routine circumstances) of DMSA (2,3 dimercaptosuccinic acid) and environmental remediation as compared with placebo and environmental remediation on children with blood lead (BPb) levels of 30-45 micrograms/dL (1.45-2.17 mumol/L). The endpoints were BPb at 1 month and 6 months after study entry. This double-blind placebo-controlled trial involved 39 children aged 2-5 years, who were randomized to one course of DMSA or placebo. The mean BPb levels of the two groups at study entry were similar, placebo group 33.0 micrograms/dL (1.59 mumol/L) and the DMSA group 34.9 micrograms/dL (1.68 mumol/L). At 1 month (the end of treatment) the mean BPb levels of the two groups were: placebo group 33.2 micrograms/dL (1.60 mumol/L) and the DMSA group 27.4 micrograms/dL (1.32 mumol/L), p = 0.16. At 6 months, the mean BPb levels were 25.1 micrograms/dL (1.21 mumol/L) for the placebo group and 28.8 micrograms/dL (1.39 mumol/L) for the DMSA-treated group, p = 0.06. Neither of these differences is statistically significant. All children with BPb, in the range studied here, should receive environmental evaluation and remediation; DMSA does not improve long-term blood lead levels. PMID- 10378090 TI - Pediatric imported malaria in New York: delayed diagnosis. AB - The records of 20 children with imported malaria admitted to Kings County Hospital between October 1987 and May 1995 were reviewed. All had a history of recent travel or immigration from a malaria endemic area (West-Africa [16], Central-America [three], and the Caribbean [one]). None of the 10 children with a travel history received appropriate malaria chemoprophylaxis. The most common symptoms and signs were daily fever, chills, and hepatomegaly. Diagnosis was delayed in seven children who were initially felt to have pharyngitis or viral syndrome. Common laboratory findings were anemia and thrombocytopenia. P. falciparum was identified in 70% of the patients. Other species were P. malariae and P. vivax. Complications occurred in six children, hyponatremia in five, seizures in three, and cerebral malaria in one patient. The high incidence of chloroquine-resistant malaria makes chemoprophylaxis difficult in children. The clinical presentation of malaria is nonspecific, and diagnostic delays occur, so a high index of suspicion is needed in children with a travel history. PMID- 10378091 TI - Ask, advise, assist: pediatricians and passive smoke exposure. AB - The objectives of this study were to determine: (1) how frequently pediatricians obtain a history of passive smoke exposure (PSE), (2) what type of advice regarding PSE they offer and how frequently they offer it, and (3) what methods and what assistance they believe would be useful to reduce PSE. A random sample of 1,000 US members (GEN) of the American Academy of Pediatrics (AAP) and all 724 members of the AAP sections of pulmonology, otolaryngology, and allergy (SPECS) were sent a questionnaire. Seven hundred fifty-five usable questionnaires were returned. Ninety-six percent of 321 general pediatricians obtained a PSE history at least "sometimes" but were much more likely to "always" do so when seeing a patient with asthma (87%) or recurrent otitis media (56%) than during well-child visits (41%) (p < 0.0001). Ninety-eight percent of pulmonologists and 95% of allergists "always" obtained a PSE history from parents of their asthmatic patients as compared with generalists who reported doing so 87% of the time (p = 0.0004). Fifteen percent of GEN gave specific assistance to parents with smoking cessation such as referral to an internist or family practitioner or a community agency or initiating a smoking cessation program themselves, whereas 85% gave only nonspecific advice such as, "don't smoke around the child," or "quit smoking." Reasons for not initiating a cessation program included lack of skills (38%) or time (36%) or a belief that it was "not their responsibility" (13%). Pediatricians indicated that brochures for parents that describe the hazards of PSE and contain specific information regarding how to refer to community smoking cessation programs would be of most use to them in helping parents reduce PSE to their children. Pediatricians frequently ask about PSE and advise reducing it but seldom assist parents with specific advice regarding effective methods to quit smoking. PMID- 10378092 TI - Parent-child relationship disorders: what do the Child Vulnerability Scale and the Parent Protection Scale measure? AB - While scales exist to aid clinicians in the assessment of parent-child relationship disorders, there are minimal data regarding their clinical usefulness. This study examined the ability of parent responses to questions regarding the child's health, behavior, and development to predict total scores on the Child Vulnerability Scale and the Parent Protection Scale. Of 120 potential participants, 103 parents (92% mothers, 69% white, 54% married, 44% lower socioeconomic status) with children aged 2-5 years completed the Child Health and Family Functioning Questionnaire. Logistic regression yielded a correct prediction rate of 77% for perceived child vulnerability and 76% for parental overprotection. Our data provide support for the independence of parental perceptions of increased child vulnerability and parental overprotection as well as for the content validity of the Child Vulnerability Scale and the Parent Protection Scale. PMID- 10378093 TI - Group A beta-hemolytic streptococcal pharyngitis in preschool children aged 3 months to 5 years. AB - The authors describe a prospective study of 420 patients aged 3 months to 5 years who presented to a primary pediatric clinic owing to fever > or = 38 degrees C and signs of pharyngitis and were not treated with antibiotics in the preceding week. Throat cultures and blood antistreptolysin O (ASO) titers were examined. In group A beta-hemolytic streptococcus (GABHS)-positive patients, a second ASO sample was obtained 2-3 weeks later. Positive throat cultures to GABHS were found in 61 of 415 patients (14.7%) (five patients were lost to follow-up). Thirty three of these (54.1% of the culture-positive group and 8% of the total study group) had the streptococcal infection with elevated ASO titers. The incidence of both true infection and carrier state gradually increased with age. Nevertheless, true streptococcal pharyngitis was found even in patients younger than 1 year and its percentage related to carriers did not increase with age and was > or = 50% in all age groups up to 4 years. The authors conclude that true GABHS pharyngitis may present in the first year of life. PMID- 10378094 TI - Group A streptococcal pharyngotonsillitis in children less than 2 years of age- more common than is thought. PMID- 10378095 TI - Group A streptococcal infections in toddlers. PMID- 10378096 TI - Cryptosporidia enterocolitis in an immunocompetent infant treated with paromomycin. PMID- 10378097 TI - Fetal cholelithiasis: a prospective study of incidence, predisposing factors, and ultrasonographic and clinical features. PMID- 10378098 TI - Comparison of an intraoral camera with colposcopy in sexually abused children. PMID- 10378099 TI - Urokinase in the management of complicated parapneumonic effusions in children. PMID- 10378100 TI - Heterogeneous responses of cell Ca2+ in human airway epithelium. AB - The Ca(2+)-mobilizing actions of adenosine 5'-triphosphate (ATP), bradykinin, and histamine were compared in phenotypically distinct human nasal epithelial (HNE) cell types and as a function of time in cell culture. Single-cell measurements of intracellular free Ca2+ (Ca2+i, Fura-2 fluorescence) were recorded in ciliated cells 1-2 days in primary culture, and in nonciliated cells 1-2 days (keratin 14 positive) or 4-5 days (keratin 18-positive) after seeding. No difference in basal Ca2+i was noted between ciliated and nonciliated cell preparations. For ciliated and nonciliated cells studied 1-2 days in culture, ATP, bradykinin, and histamine elicited a cytosolic Ca2+ response in 100% of the cells examined. For nonciliated HNE cells maintained 4-5 days in culture, ATP (10(-4) M) increased cytosolic Ca2+ in all cells tested, but only 85% of the cells responded to bradykinin (10(-5) M) addition, and 65% to histamine (10(-4) M) stimulation. In terms of the absolute change of Ca2+i (delta Ca2+i, peak-basal value), the efficacy was ATP > bradykinin > histamine for the 3 HNE cell preparations. However, the delta Ca2+i in response to agonists was smaller in nonciliated HNE cells studied 1-2 days or 4-5 days in culture as compared to the ciliated cell preparation. Thapsigargin (300 nM), an agent that mobilizes Ca2+i, was equally effective in raising cytosolic Ca2+ in nonciliated (1-2 days and 4-5 days in culture) and ciliated HNE cells. These data show that ciliated cells consistently respond to all agonists, whereas the cytosolic Ca2+ response to ATP, bradykinin, and histamine in nonciliated cells was quantitatively reduced at a comparable time period (1-2 days) and became smaller and less frequent in nonciliated cell preparations maintained 4-5 days in culture. These results demonstrate time-dependent differences in the magnitude and frequency of cytosolic Ca2+ responses to certain agonists, strongly indicating that measurements of Ca2+i in HNE cells must account for the heterogeneity of the cell types and the time cells are maintained in primary culture. PMID- 10378101 TI - Effect of acute glucose depletion following glucose excess on surfactant phospholipid synthesis in developing fetal lung. AB - Exposure to high glucose and insulin inhibits surfactant synthesis in vitro. We have demonstrated that fetal lung insulin receptor tyrosine kinase (TK) activity is downregulated after culture in high glucose plus insulin, with a resultant decrease in glucose uptake. To see whether relative substrate depletion following substrate excess would further diminish surfactant synthesis, 20-day fetal rat lung explants were initially cultured for 44 hours in media containing 100 mM glucose with or without 0.1 U/mL insulin, followed by a 4-hour pulse in 10 mM glucose +/- insulin or 100 mM glucose +/- insulin, after which the rate of choline incorporation into phosphatidylcholine (PC) or disaturated PC was measured. Choline incorporation was significantly lower after a 4-hour pulse in low glucose (+/- insulin) than under continuing high glucose (+/- insulin) conditions (P < .01). To determine the time required for reversal of TK downregulation in lung explants, insulin receptor TK activity was assayed after 44 hours in high glucose + insulin, followed by an additional 4, 8, 12, or 24 hours in either 10 mM glucose or continued 100 mM glucose + insulin. After 44 hours in 50 mM or 100 mM glucose + insulin, TK activity was significantly decreased (68 +/- 9% and 57 +/- 9% of control; P < .01). When explants cultured in 100 mM glucose + insulin for 44 hours were subsequently placed in 10 mM glucose for an additional 4 or 8 hours, TK activity remained significantly downregulated (70.2 +/- 7.0% and 84.9 +/- 5.5% of control, respectively, P < .05) compared to explants cultured in low glucose throughout. However, after 12 or 24 hours in low glucose, TK activity was no longer significantly different from control values (106.5 +/- 2.1% and 106.0 +/- 19.6%, respectively). Culture of type II cells for 44 hours in 25 mM glucose + insulin, followed by an additional 4 or 24 hours in either low glucose (5.5 mM) + insulin or high glucose (25 mM) + insulin yielded similar results: TK activity was decreased 20-30% by culture in 25 mM glucose + insulin conditions (P < .05) and this downregulation continued for 4 (but not 24) hours after switching to lower glucose conditions (P < .05). Continued receptor downregulation during a period of relative substrate deprivation may adversely affect surfactant synthesis, as in some infants of diabetic mothers who experience hypoglycemia (following intrauterine hyperglycemia) in the immediate postnatal period. PMID- 10378102 TI - A rat model presenting eosinophilia in the airways, lung eosinophil activation, and pulmonary hyperreactivity. AB - The aim of this study was to examine antigen-induced lung cell migration, eosinophil activation, and pulmonary reactivity of Wistar rats exposed to a new sensitization technique. The animals were sensitized with a single subcutaneous implant of a fragment of heat coagulated hen egg white and challenged 21 days later with an intratracheal injection of heat-aggregated ovalbumin (EWI). For comparison, another group of rats were sensitized by an intraperitoneal injection of ovalbumin in alum as adjuvant, with one booster on day 14 and challenge on day 21 post immunization (OVA/AL). Twenty-four hours after antigen challenge, the EWI group presented a higher number of eosinophils in the bronchoalveolar lavage (BAL) (4.85 +/- 1.43 x 10(6)) than the OVA/AL group (0.2 +/- 0.06 x 10(6)) or the control group, where the level of eosinophils were essentially undetectable. Levels of eosinophil peroxidase activity were increased in the cell-free BAL and homogenates of lung tissue in the EWI group (12.10 +/- 2.97 mg/mL and 36.14 +/- 7.21 ng/mg, respectively), but not in the OVA/AL group (4.83 +/- 1.4 ng/mL and 11.95 +/- 2.54 ng/mg, respectively), as compared with controls (5.16 +/- 1.65 ng/mL and 12.13 +/- 1.74 ng/mg, respectively). Thromboxane B2 levels were also increased in the BAL of EWI group (2.89 +/- 0.54 ng/mL) but not the OVA/AL group (1.13 +/- 0.23 ng/mL) as compared with controls (1.14 +/- 0.19 ng/mL). In contrast, the levels of prostaglandin E2 in the BAL were increased in both groups (456.4 +/- 11.8 pg/mL in the EWI group and 303.5 +/- 31.7 pg/mL in the OVA/AL group) as compared with controls (205.7 +/- 29.7 ng/mL). Moreover, only the EWI group developed increased pulmonary reactivity to serotonin (around two-fold), 24 hours after antigen challenge. The extent of lung eosinophil migration and activation and the pulmonary hyperreactivity induced by this novel sensitization procedure without adjuvants represents a significant improvement over existing experimental models of asthma. PMID- 10378103 TI - Perfluorochemical liquid-enhanced adenoviral vector distribution and expression in lungs of spontaneously breathing rodents. AB - Perfluorochemical (PFC) liquids have been shown to improve gas exchange and lung compliance in models of lung injury. We reasoned they may also be useful as a vehicle for gene transfer by improving transgene distribution throughout the lung as well as increasing total transgene expression. We have developed a model for PFC liquid use in spontaneously breathing rodents that obviates the need for intubation and ventilation. Intratracheal instillation of the adenoviral vector Adlac-Z resulted in patchy distribution of beta-galactosidase (beta-gal) activity as demonstrated using X-gal histochemistry. In contrast, in rats instilled with Adlac-Z followed by instillation of PFC liquid, more uniformly distributed and increased beta-gal activity was observed. Activity in distal airway and alveolar epithelium was particularly increased. Quantitative measure of beta-gal activity in lung homogenates demonstrated a 3- to 6-fold increase in total activity in lungs of rats receiving Adlac-Z and PFC liquid compared to animals receiving Adlac-Z alone. These studies show that PFC liquids can enhance both the distribution and the total amount of transgene expressed following adenoviral mediated vector transfer to lungs during spontaneous breathing. Use of PFC liquids may increase the efficacy of gene transfer strategies for treatment of cystic fibrosis and other lung diseases. PMID- 10378104 TI - Human tracheobronchial deposition and effect of a cholinergic aerosol inhaled by extremely slow inhalations. AB - Ten subjects inhaled the same amounts of cholinergic aerosol of a mass median diameter (MMD) of 7.7 microns in a normal provocation test and in a test with extremely slow inhalations (ESI). This new technique using ESI and large droplets/particles gives a high degree of deposition in small ciliated airways which cannot be obtained by using small particles. The purpose was to compare measured effects with calculated doses of the aerosol in large and small ciliated airways. The effect on large airways was measured by airway resistance (R(aw)), and the effect on small airways was measured by the phase III slope of single breath nitrogen test (N2-delta). Mouth and throat deposition was calculated from human experimental data, and deposition of the cholinergic aerosol into large and small airways was calculated, using a computerized lung model. The study showed that the extremely slow inhalation caused a larger effect on R(aw) and tendency to a larger effect on N2-delta compared to the effect in the normal provocation. Deposited dose in the large airways, in percent of inhaled dose, was calculated to be 25-33% for normal inhalation and 20-24% for ESI. Calculated deposited dose in the small airways (bronchioles; generations 12-16) was 1.8-3.4% for normal inhalation and 18-25% for ESI. For large airways a stronger effect was induced by ESI, perhaps by the more uniform distribution of particles within each generation, compared to normal inhalations when particles deposit near the bifurcations. Concerning the small airways, N2-delta did not differ significantly between normal and ESI provocations, indicating that they did not react much on cholinergic exposure. We believe that our approach using ESI for small airway deposition of a nebulized aerosol can be of value for estimating the effects of various substances on large and small airways. PMID- 10378105 TI - Lung deposition and extremely slow inhalations of particles. Limited effect of induced airway obstruction. AB - Studies of lung deposition and clearance have focused on the large airways. Still, lung diseases affect also the small airways. We have developed a method for selective particle deposition in the smallest ciliated airways. Eight healthy subject inhaled 6-micron radiolabelled test particles on 3 occasions at 0.05 L/s and retention was measured for 72 hours. At one occasion, the subjects inhaled the particles at a normal airway resistance. At a second occasion, a 2-3-fold increase in airway resistance was induced by a cholinergic provocation before inhalation of the particles. At a third occasion, a corresponding provocation was induced after inhalation of the particles. The percentage lung depositions were 76 +/- 7, 68 +/- 7, and 73 +/- 8 (mean +/- SD) for "normal airway resistance," "provocation before," and "provocation after" exposures, respectively. The lower value for the "provocation before" exposure was probably a result of increased mucociliary clearance, due to cholinergic stimulation, before the first measurements of radioactivity. The retentions at 24 hours were 51 +/- 7, 52 +/- 9, and 51 +/- 8 in percent of initial lung deposition for "normal airway resistance," "provocation before," and "provocation after" exposures, respectively. We conclude that our inhalation technique is useful in studying conditions in the bronchioles, as deposition is rather independent of airway resistance. PMID- 10378106 TI - Anxiety in primary care depression: how does it lead to poor outcomes and what can we do about it? PMID- 10378107 TI - DSM-IV somatoform disorders: do we need a new classification? PMID- 10378108 TI - The ethics of research involving memories of trauma. PMID- 10378109 TI - Does a coexisting anxiety disorder predict persistence of depressive illness in primary care patients with major depression? AB - We assessed whether a coexisting anxiety disorder predicts risk for persistent depression in primary care patients with major depression at baseline. Patients with major depression were identified in a 12-month prospective cohort study at a University-based family practice clinic. Presence of an anxiety disorder and other potential prognostic factors were measured at baseline. Persistent depressive illness (major depression, minor depression, or dysthymia) was determined at 12 months. Of 85 patients with major depression at baseline, 43 had coexisting anxiety disorder (38 with social phobia). The risk for persistent depression at 12 months was 44% greater [Risk Ratio (RR) = 1.44, 95% confidence interval (CI) 1.02-2.04] in those with coexisting anxiety. This risk persisted in stratified analysis controlling for other prognostic factors. Patients with coexisting anxiety had greater mean depressive severity [repeated measures analysis of variance (ANOVA), p < 0.04] and total disability days (54.9 vs 19.8, p < 0.02) over the 12-month study. Patients with social phobia had similar increased risk for persistent depression (RR = 1.40, 95% CI 0.98-2.00). A coexisting anxiety disorder indicates risk for persistent depression in primary care patients with major depression. Social phobia may be important to recognize in these patients. Identifying anxiety disorders can help primary care clinicians target patients needing more aggressive treatment for depression. PMID- 10378110 TI - Training primary care physicians improves the management of depression. AB - The purpose of this pretest-posttest study was to evaluate effects of a training program designed to improve primary care physicians' (PCPs) ability to recognize mental health problems (MHP) and to diagnose and manage depression according to clinical guidelines. The primary care settings were in the northern part of The Netherlands. There were eight intensive, hands-on training sessions of 2.5 hours, each of which three were targeting depression (7.5 hours). In the pretraining phase we screened 1778 consecutive patients of 17 PCPs with the 12-item General Health Questionnaire (GHQ-12) and interviewed a stratified sample of 518 patients about presence of current depression with the Primary Health Care version of the Composite International Diagnostic Interview (CIDI-PHC). PCPs registered patient's mental health (status, severity, diagnosis) and treatment prescribed. Then we trained the PCPs. In the posttraining phase, we screened a new group of 1724 consecutive patients of the same PCPs and a new stratified sample of 498 patients went through the same interview and rating procedures as patients in the pretraining phase. Knowledge about depression was assessed pre- and posttraining. PCPs' knowledge of depression improved significantly. Recognition of MHP and accuracy of depression diagnosis improved, but was not statistically significant. The proportion of patients receiving treatment according to the clinical guidelines increased significantly. It was observed that training PCPs improves the management of depression. PMID- 10378111 TI - The somatoform conundrum: a question of nosological values. AB - This paper critically reviews the values and perspectives evident in the formulation of the category of Somatoform Disorders in the widely used Diagnostic and Statistical Manual-IV (DSM-IV) [1] of the American Psychiatric Association. The conflict of values evident in the DSM Committee's elimination of causal issues and including only descriptive criteria in the final classification is emphasized. In this approach, causation was dismissed and only a description of popular clinical entities was allowed. It becomes clear that there was a conflict between causal and descriptive factors in the formation of the current classifications. The dubious logic and the inconsistencies that underlie the linking of the various diagnoses under the Somatoform Disorder (SD) rubric is presented. A heuristic model of the interaction of psychological and somatic variables is described. These variables act in a causal fashion and should be the basis for any nosological system of the SDs. There is ample evidence to support the importance of causal factors in the development of SDs, and the current descriptive nomenclature does not do justice to the rapidly growing field of Psychosomatic Medicine. A new scheme is proposed, using the presence or absence of stress as the connecting link for the diagnoses offered. The old nosology enhances the perpetuation of the mind-body dichotomy. Stress, representing the way in which the environment impinges on the human being, allows the mind-body dichotomy to be eliminated. The preferred synthesis of mind-body interaction as partly the result of experience in the environment becomes the basis for the new nosology. PMID- 10378112 TI - Assessing the ethical costs and benefits of trauma-focused research. AB - Although scientists and members of Institutional Review Boards must balance the needs of investigators and participants in research, virtually no evidence is available to inform this decision making. This study examines the frequency and correlates of adverse reactions and adequacy of informed consent among 1174 women in an HMO who completed a trauma-focused health survey, and a subset of 252 women who later completed a trauma-focused research interview. Despite the sensitive content, the majority of women participants found participation in the interview and the questionnaire study to be a positive experience. Although a small number of women, particularly those with a history of maltreatment, underestimated the level of upset they would subsequently experience, the majority still did not regret participating, indicating that informed consent procedures were adequate, with a large proportion reporting immediate perceptions of personal gain. Finally, the cost-benefit ratio appears stable 48 hours post-interview, with some minor fluctuations. Overall, these results suggest that research on childhood victimization is well tolerated by women who participate. Though a small number of women may be disturbed by these investigations, in general, adverse reactions appear less common than previously anticipated. PMID- 10378113 TI - Follow-up consultation billing and documentation. AB - Frequently, bills are not submitted for follow-up visits for patients who have been evaluated psychiatrically on medical-surgical services. There often is confusion regarding which procedure codes are most appropriate to use in billing. To help the consultant understand the documentation requirements for various procedure codes, information from several sources was synthesized and distilled. This paper should help minimize documentation errors and maximize reimbursement for clinical services. The authors have reviewed available billing choices, and clarified the documentation requirements for different procedure codes according to Medicare regulations. PMID- 10378114 TI - Disclosure of true diagnosis in Japanese cancer patients. AB - Full disclosure of medical diagnosis to cancer patients in Japan remains controversial. Some physicians in Japan believe that full disclosure may affect the outcome of treatment, create stress and psychiatric problems, or lead to suicide. Although the trend toward full disclosure is increasing in Japan, approximately 70% of current cancer patients are still not fully informed of their condition. In this study, the authors examined the psychiatric status and effects of full disclosure among 100 otolaryngology patients at Tokai University Hospital (50 with benign diseases, 50 with malignancy) using major depression and adjustment disorders criteria of the DSM-III-R Structured Clinical Interview (SCID). This demonstrated that 15 of 50 (30%) patients with benign diseases and 23 of 50 (46%) patients with malignant diseases met the criteria for depression and adjustment disorder; 29 of the 50 patients (58%) with malignant cancer were not informed of their true condition, according to the wishes of their families (21 were fully informed). The prevalence rate of psychiatric disorders was 42.9% among the informed group and 48.3% among the uninformed group. These findings suggest that concealing the true diagnosis was not related to the presence of psychiatric disorders in Japanese cancer patients. PMID- 10378115 TI - The disruptive behavior disorders in the psychiatric emergency service. AB - Many children and adolescents presenting to the Psychiatric Emergency Service (PES) are diagnosed with the Disruptive Behavior Disorders (DBD), Conduct Disorder (CD), and Oppositional Defiant Disorder (ODD). Sometimes it may be difficult to reliably diagnose these disorders in the PES setting. Given these limitations, a large database of 6 years of PES visits showed 314 patients with DBD compared with 1625 without DBD. More DBD patient visits required the intervention of police and/or the mobile crisis team. These patients are more likely to have additional diagnoses of depression and attention deficit hyperactivity disorder, be in current treatment, or involved with court or the correctional system. They are less frequently referred by other emergency services such as medical ERs. DBD patients do not require emergency medication or psychiatric hospitalization any more frequently than other youngsters presenting to the PES. In the PES setting there is little differentiation between the CD and the ODD population. A more detailed study of the presenting symptomatology of the DBD vs non-DBD patients revealed that DBD patients showed over twice as many disruptive behavior symptoms. Fights and defiance were present significantly more frequently than in controls, with a trend toward increased frequency of bullying and stealing. The clinical and public mental health implications of these findings are discussed. PMID- 10378116 TI - Laboratory findings in acute schizophrenia. Relevance to medical management on emergency admission. AB - This study documents the prevalence of abnormal laboratory findings in schizophrenic patients who were admitted, because of acute disease, to a psychiatric intensive care unit in Japan. Patient laboratory data were evaluated retrospectively prior to treatment. Of 259 male acute schizophrenic patients (ICD 10: F2 group), nearly 10% were dehydrated, 33% had hypokalemia and leukocytosis, and 66% showed elevated serum muscle enzymes. This prevalence was statistically significant compared with that of psychiatric outpatients (F1 group). In addition, these medical problems in the F2 group were as frequent as those in the F1 group, i.e., alcohol and/or psychoactive substance abusers (ICD-10), although the problems in the F2 group occurred less often than in the F1 group. Current medication, obvious complications, or the presence of alcohol and/or psychoactive substance abuse in the F2 group were not major causes of these results. The medical problems significantly improved after 8 hours of fluid therapy. These findings strongly suggest the significance of medical management for acute schizophrenic patients on emergency admission as well as for alcohol and/or psychoactive substance abusers. PMID- 10378117 TI - Calcium binding proteins immunohistochemistry and identification of neurons in the mammalian pineal gland of the African giant rat: Cricetomys gambianus. AB - The presence of true neurons in the rodent pineal gland is still a matter of controversy. In this work, by using immunohistochemistry with five antibodies against calcium-binding proteins (calbindin-D28k, calretinin, calmodulin, neurocalcin and S-100 beta) and Cricetomys gambianus, a rodent belonging to Muridae family living in Africa, we were able to illustrate the presence of neurons in the pineal gland. Anti-calbindin-D28k and anti-calretinin labelled neurons belonging to two neural ganglia. One ganglion was localized in the anterior part of the gland near the pineal stalk and the other one in the posterior portion of the organ. Immunoreactive neurons are medium in size (15-20 microns) and have long thick processes running towards the stalk. Calretinin and calbindin-D28k positive neurons stained with different intensities. Thin processes were detected by anti-calretinin whereas thick processes were preferentially calbindin-D28k positive. Neurocalcin labelled a few smaller neurons and many thin processes within the ganglion. Calmodulin could not be detected immunochemically. Within the ganglia many astrocytic processes were S 100 beta positive. The afferent and the efferent pathways of the pineal ganglia remain to be elucidated. PMID- 10378118 TI - Effects of strontium ions on contraction and action potential in rabbit papillary muscles. A comparison with effects of tetraethylammonium ions. AB - The effects of Sr2+ on contraction and action potential were studied in rabbit papillary muscles and compared with effects of tetraethylammonium (TEA+). The membrane potential was measured with KCl-filled microelectrodes and the contraction was simultaneously recorded using a mechanoelectrical transducer. A partial (90%) substitution of extracellular Ca2+ (Ca2+e) by Sr2+ produced stimulation frequency-dependent prolongation of the action potential (AP) with a dominant phase "plateau" as well as prolongation of the contraction. At low frequencies where the AP prolongation was well pronounced, the contraction became biphasic. The effect of Sr2+ on both AP and contraction was blocked by nifedipine (10 mumol/l) or by increasing Ca2+e. Ryanodine suppressed the early contraction component only. AP was prolonged to a similar extent and in the same frequency dependent manner by TEA+ (20 mmol/l). Despite similar AP configuration, no biphasic contraction developed in the presence of TEA+. High Ca2+e (10 mmol/l) or low Na+e (70 mmol/l) suppressed the TEA+ effect on AP. The data indicate that the two components of the biphasic contraction are of different origin; the early one is activated by activator cation released from the sarcoplasmic reticulum while the late one results from the Sr2+ entry across the sarcolemma via L-type Ca2+ channels. PMID- 10378119 TI - Effects of progesterone and estradiol benzoate on glutathione dependent antioxidant enzyme activities in the brain of female rats. AB - The activities of glutathione dependent antioxidant enzymes were measured in subcellular fractions of whole brain homogenates prepared from ovariectomized (OVX) female rats, untreated or treated 2 h or 24 h prior to sacrifice with a single dose of 2 mg progesterone (P) or 5 micrograms estradiol benzoate (EB). Glutathione peroxidase (GSH-Px) activity was not changed following systemic administration of EB, but P increased GSH-Px in the brain of OVX rats 24 h after the treatment. The activity of glutathione reductase (GR) was suppressed by EB short time, only 2 h following treatment, whereas P increased the enzyme activity 24 h after treatment. On the other hand, the activities of catalase (CAT) and glutathione-S-transferase (GST) were not changed following systemic administration of EB or P. The present work was carried out to study the involvement of ovarian steroids, especially P, in the control of GSH-Px and GR activities, and our results suggest that oxidative stress in the brain of female rats may be modulated by the level of progesterone. PMID- 10378120 TI - Differential expression of regulatory proteins in L1210/VCR cells with multidrug resistance mediated by P-glycoprotein. AB - Phosphorylation of P-glycoprotein (PGP) by some protein kinases may play an important role in the regulation of its drug transport activity, and may also be important for the development of multidrug resistance (MDR) phenotype. In the present study we investigated the expression of three groups of mitogen-activated protein kinases (MAPKs). The expression of ERKs, SAPK/JNKs and p38-MAPK was studied at the protein level in sensitive (L1210) and multidrug resistant (L1210/VCR) cells. The expression of ERKs in multidrug resistant cells did not differ from those observed in parental sensitive cells. On the other hand, the development of multidrug resistance phenotype in L1210/VCR cells was associated with increased expression of cytosolic p38-MAPK and also proteins of 90 and 130 kDa that react with antibody specific for SAPK/JNKs. The expression of the proteins mentioned was stimulated above all in conditions when vincristine was present in cultivation medium and the stimulation of transport activity of PGP was necessary for the cell survival. The development of multidrug resistance phenotype in L1210/VCR cells was not associated with significant changes in expression of several heat-shock proteins (hsp25, hsp60, hsp70, hsp90). The levels of these proteins were comparable in sensitive L1210 and resistant L1210/VCR cells, and vincristine did not influence the expression of heat-shock proteins in resistant cells. PMID- 10378121 TI - Depression of acetylcholinesterase synthesis following transient cerebral ischemia in rat: pharmacohistochemical and biochemical investigation. AB - The effect of transient cerebral ischemia on acetylcholinesterase (AChE) synthesis was studied in rats by a modified pharmacohistochemical method. The procedure involved in vivo irreversible inhibition of AChE by administration of the inhibitor diisopropyl fluorophosphate (DFP; 1.2 mg/kg b.w., i.m.) 1 h before 30 min forebrain ischemia (the four-vessel occlusion model). At the onset of ischemia, 70-75% of AChE was inhibited in the brain. Recirculation was followed by histochemical and biochemical investigations of newly synthesized AChE in the striatum, septum, cortex and hippocampus. Control sham-operated animals were treated with the same dose of DFP. For correlation, rats not treated with DFP were subjected to the same ischemic procedures and investigated simultaneously. In these rats, significant decrease in AChE activity was found in the striatum, septum and hippocampus during 24 h recirculation. In DFP treated rats, ischemia markedly depressed resynthesis of AChE; after 4 h recirculation, AChE activity was decreased by 45-60% in all investigated areas in comparison with controls and the AChE histochemistry showed only slightly stained neurons in the striatum and septum. Twenty-four hours after ischemia, these neurons were densely stained and the increase in AChE activity indicated a partial recovery of the enzyme synthesis. These results suggest that the depression of AChE synthesis after forebrain ischemia is probably transient, not accompanied by cholinergic neuron degeneration. PMID- 10378122 TI - The parametric pump mechanism in separation of components in heterogeneous systems. I. Macroscopic distributed systems. AB - A dynamic method is proposed for the separation of the electrolyte components using a parametric pump with an ion exchange column. It was studied experimentally and described mathematically. The parametric separation of mixtures is based on interactions of two oscillating fields with a heterogeneous system containing two phases, a liquid and a solid one, the components of the mixture being able to redistribute between the phases. The field of mechanical force is responsible for cyclic relative displacement of the phases, and synchronously changing temperature causes redistribution of the components between them. This results in sodium and potassium fluxes opposite in direction which in turn leads to accumulation of sodium and potassium in opposite end cells. PMID- 10378123 TI - Serum level of IgG autoantibodies against oxidized low density lipoproteins and lag-phase of serum oxidation in coronary heart disease--inverse correlation. AB - High affinity IgG autoantibodies (ABs) against oxLDLs and lag-phase of serum oxidation were tested in patients with coronary heart disease (CHD). Fifty one (37 M/14 F) patients with CHD defined as Q-wave myocardial infarction and/or stenosis of more than 50% and 51 (34 M/17 F) healthy blood donors as controls participated in this study. LDLs were isolated by gradient ultracentrifugation and oxidized with CuSO4. The modified LDLs (oxLDLs) or native LDLs (nLDLs) were used as antigens in an enzyme immunoassay (ELISA) to detect IgG ABs in both groups. The serum was oxidized by CuSO4 and the oxidation was monitored spectrophotometrically at lambda = 234 nm to follow the formation of conjugated diens. The lag-phase (in minutes) is the interval between the addition of CuSO4 to the serum and the beginning of extensive oxidation (increasing absorbance at 234 nm). The concentrations of total cholesterol, triglycerides, HDL-cholesterol, apo-A and apo-B were measured as well. The mean level of ABs against oxLDLs (expressed as optical density units) was 0.590 +/- 0.330 in CHD-patients vs 0.244 +/- 0.200 in controls (p < 0.001). The lag-phase in minutes was 47.00 +/- 27.19 in CHD-patients and 80.23 +/- 26.30 in controls (p < 0.001). A negative correlation between ABs levels and lag-phase was established in CHD-patients (r = -0.69, p < 0.001) and controls (r = -0.62, p < 0.001). A poor correlation was established between ABs levels or lag-phase, on one hand, and other measured parameters. In conclusion, the lag-phase of serum oxidation by Cu2+ could be informative for LDL susceptibility to modification and the extent of consequent humoral immune response. PMID- 10378124 TI - Is antimicrobial therapy of value for all children with acute otitis media? PMID- 10378125 TI - A primer on anaerobic bacteria and anaerobic infections for the uninitiated. PMID- 10378126 TI - Use of newer quinolones for the treatment of intraabdominal infections: focus on clinafloxacin. PMID- 10378127 TI - Dynamics of interleukin-1 production in middle ear fluid during acute otitis media treated with antibiotics. AB - In an ongoing prospective study, IL-1 concentrations were measured in 78 children (aged 3-36 months) with acute otitis media receiving antibiotics. Middle ear fluid IL-1 concentrations were determined using ELISA kits. Ninety-eight middle ear fluid samples were obtained by tympanocentesis at enrollment (day 1) and 43 samples were collected on days 4-5. Ninety-two pathogens were isolated in 77/98 samples obtained on day 1: 55 Haemophilus influenzae, 34 Streptococcus pneumoniae, 2 Moraxella catarrhalis and 1 Streptococcus pyogenes. Among 37 paired samples initially culture-positive, eradication of the pathogen was achieved on day 4-5 in 20 while pathogens were still present in 17. On day 1, IL-1 was detected in 61/77 (79%) culture-positive samples vs 9/21 (43%) culture-negative ones (P = 0.003). The mean +/- SD middle ear fluid concentration of IL-1 on day 1 was significantly higher in culture-positive (316 +/- 508 pg/ml) than in culture negative samples (111 +/- 245 pg/ml) (P = 0.01). When paired samples were evaluated, IL-1 decreased on days 4-5 in 13/20 (65%) ears where bacterial eradication was achieved, but also in 11/19 (58%) with persistent or new infection. The mean IL-1 concentrations decreased on days 4-5 in the 20 samples from ears where bacterial eradication was achieved (330 +/- 460 vs 118 +/- 294 pg/ml, P = 0.1) but also in the 17 samples where it was not (465 +/- 660 vs 232 +/- 289 pg/ml, P = 0.02). No significant differences were found between day 1 and days 4-5 in the mean IL-1 concentrations measured in patients with H. influenzae vs S. pneumoniae or concomitant H. influenzae and S. pneumoniae. It was concluded that: 1) IL-1 was detected in the middle ear fluid of most patients with acute otitis media; 2) significantly higher IL-1 concentrations were found in patients with culture-positive than in those with culture-negative acute otits media; 3) IL-1 concentrations decreased on days 4-5 of antibiotic therapy, whether the pathogen was eradicated or not. PMID- 10378128 TI - Serodiagnosis of neuroborreliosis: comparison of reliability of three confirmatory assays. AB - The sensitivity and specificity of three confirmatory assays for the serodiagnosis of neuroborreliosis were investigated. Samples from 96 patients with proven neuroborreliosis, 80 healthy volunteers, 20 patients with neurosyphilis and 20 patients with recent infections with Epstein-Barr virus (EBV) were tested for borrelial antibodies by immunoblotting, Borrelia burgdorferi sensu lato sonicate EIA following pre-absorption of cross-reactive antibodies (Abs-EIA) and by a so-called RECO-EIA using the following recombinant borrelial proteins as antigens: a 14 kDa-internal flagellin fragment, the outer surface protein C (23 kDa) and the high molecular mass protein p83 (83 kDa). The immunoblots were evaluated according to the criteria published by Engstrom et al. and Hauser et al. An evaluation of IgM and/or IgG antibodies revealed a considerably higher sensitivity for the RECO-EIA (94%) compared to the Abs-EIA (82%, P < 0.0001). Evaluation of the immunoblot according to the criteria of Hauser was significantly more sensitive than according to the criteria of Engstrom (89 vs 51%, P = 0.0003). A higher sensitivity was demonstrated for IgM (54 vs 22%) and IgG antibodies (64 vs 24%). When both findings from RECO-EIA and immunoblotting were considered, positive findings in the first step assay (sonicate EIA without pre-absorption) were confirmed in 97% of patients. When samples were tested for IgM antibodies, the specificities of the three confirmatory assays did not differ significantly, but in the case of IgG antibodies, the immunoblot (Hauser: P = 0.013; Engstrom: P = 0.004) and the RECO EIA (P = 0.02) were more specific than the Abs-EIA. It is concluded that the immunoblot (evaluated according to Hauser) and the RECO-EIA are both suitable as confirmatory assays in the serological diagnosis of neuroborreliosis. Monoclonal antibodies are mandatory tools in the evaluation of the immunoblot. PMID- 10378129 TI - Effect of polyclonal immunoglobulins on neutrophil phagocytic capacity and reactive oxygen production in patients with gram-negative septicemia. AB - The effect of immunoglobulin (Ig) preparations on neutrophil phagocytic ability and oxidative burst in response to Escherichia coli stimulation was analyzed in 14 patients with gram-negative septicemia by an ex vivo whole blood assay using flow cytometry. In patients, neutrophils exhibited a decreased capacity to phagocytize E. coli and generate reactive oxygen products compared to healthy controls (median -68%, P < 0.01). The addition of both 7S-Ig and 19S-Ig enriched preparations in vitro resulted in a dose-dependent increase in neutrophil reactive oxygen production at concentrations of 10 g/l (median +153% and +211%, P < 0.01, respectively) and 20 g/l (median +205% and +282%, P < 0.01, respectively). A decreased neutrophil phagocytic ability was seen in patients with septicemia (median -58%) compared to healthy controls (P < 0.01). Again, the addition of 7S and 19S-Igs enhanced the phagocytic ability in a dose-dependent manner (10 g/l: median +56 and +126%; 20 g/l: median +126% and +165%, P < 0.01 for all). It can be concluded that both polyclonal Igs can increase depressed neutrophil reactive oxygen production and neutrophil phagocytosis in patients with gram-negative septicemia. PMID- 10378130 TI - Incidence of Helicobacter pylori infection in a cohort of Italian military students. AB - Prevalence and incidence of Helicobacter pylori infection among 250 Italian military students were studied using specific IgG antibodies. Subjects susceptible at enrollment were evaluated during a 10-month follow-up period, when two serum samples were collected after 5 and 10 months, respectively. Samples were also analyzed for anti-CagA (a protein associated with virulent H. pylori strains) IgG antibodies. Finally, spectrotypic analysis by isoelectric focusing and reverse blotting (IEFRB) was performed in the majority of positive samples. Forty-three out of 250 (17.2%) were positive at the time of enrollment, a seroprevalence rate very similar to that observed in a larger Italian military population 5 years earlier. Among the 207 susceptible subjects, two seroconverted at 5 months after enrollment; they were still positive at the end of follow-up (incidence rate of 1.16 per 100 person/years of exposure). This data suggests a spread of H. pylori in the Italian military population that is not negligible. Nearly all anti-H. pylori-positive subjects were also CagA antibody positive (agreement percentage: 97.6%; K = 0.91), suggesting that the large majority of H. pylori strains were of the virulent type. Sixty-four percent of positive sera presented an oligoclonal spectrotype, which seems to be a hallmark of humoral immune response to H. pylori. PMID- 10378131 TI - Risk factors for hospital-acquired urinary tract infection in a large English teaching hospital: a case-control study. AB - About 10% of patients in hospital develop a hospital-acquired infection (HAI); the most commonly affected site is the urinary tract. Many studies have examined risk factors for HAI but few have adjusted for confounding and interaction. We performed a prospective case-control study on six acute wards of a busy English teaching hospital to assess risk factors for hospital-acquired urinary tract infection (HAUTI). Over a 2-year period, 136 cases were identified (2.8% of all patient episodes) along with 408 controls. Multiple logistic regression revealed that female sex, increased length of stay, elective admission, surgical operation, and transurethral and repeated intermittent catheterization were all significant independent risk factors for HAUTI. However, specialty of admission was also a significant risk factor when added to the model and, under these conditions, only length of stay and catheterization also remained significant. We detected significant interactions suggesting that the risk of HAUTI is maximal among women undergoing elective surgery, especially those who are catheterized; however, the overall risk of HAUTI among patients admitted electively was greater than for patients admitted as emergencies. PMID- 10378132 TI - Azithromycin: single 1.5 g dose in the treatment of patients with atypical pneumonia syndrome--a randomized study. AB - An open comparative study was undertaken in order to assess the efficacy and safety of a single dose of azithromycin in the treatment of community-acquired atypical pneumonia. A total of 100 adult patients with atypical pneumonia syndrome were randomized to receive 1.5 g of azithromycin as a single dose, or 500 mg once daily for 3 days. The presence of Mycoplasma pneumoniae, Chlamydia pneumoniae, Chlamydia psittaci, Coxiella burnetii, and Legionella pneumophila infection was diagnosed by serological tests. Control clinical examinations were performed 72 h, 10-12 days and 4 weeks after treatment initiation. Among 96 patients (48 in each group) who were evaluable for clinical efficacy M. pneumoniae infection was confirmed in 24, C. pneumoniae in nine, C. psittaci in five, C. burnetii in six, and L. pneumophila in five. Forty-seven patients (97.9%) in each group were cured. Side effects were observed in two patients in the single-dose group, and one patient in the 3-day group. In conclusion, a single 1.5 g dose of azithromycin may be an alternative to the standard 3-day azithromycin regimen in the treatment of outpatients with atypical pneumonia syndrome. PMID- 10378133 TI - Placental transfer of maternal rubella antibodies to full-term and preterm infants. AB - Premature infants are vulnerable to infections, partly because of the low transplacental transfer of maternal antibodies. The present study investigated the placental transfer of maternal rubella-specific antibodies to full-term and preterm infants. The study group consisted of 133 healthy, native Israeli mothers and their 159 newborns. Of these, 69 were full-term infants (gestational age > 37 weeks) of 69 mothers, and 90 were preterm infants (gestational age < 35 weeks) of 64 mothers. Antibody titers against rubella were measured in maternal and umbilical cord blood samples by hemagglutination inhibition and microneutralization techniques. There was no significant difference in the level of protection and in geometrical mean titers by hemagglutination between the full term and preterm groups. Conversely, significant differences in geometric mean titers of neutralizing antibodies were found between full-term and preterm infants, e.g., 65.9 and 39.8, respectively (P < 0.001). Very low birth weight preterm infants are at greater risk of rubella infection during the first year of life, due to the diminished transfer of neutralizing maternal antibodies. Therefore, earlier vaccination of this group may be beneficial. PMID- 10378134 TI - Resistance to third-generation cephalosporins in adult gram-negative bacillary meningitis. AB - Ninety-three patients with gram-negative bacillary meningitis (GNBM) were identified at Kaohsiung Chang Gung Memorial Hospital, over a period of 12 years. Among them, eight showed resistance to third-generation cephalosporins, accounting for 9% of the total GNBM cases and 29% of the postneurosurgical GNBM cases. The resistant pathogens included Acinetobacter baumannii, Klebsiella pneumoniae, Citrobacter freundii and Morganella morganii. These eight patients, six males and two females aged 18-61 years, all had nosocomially acquired meningitis associated with head trauma and/or postneurosurgical states. Six patients received imipenem/cilastatin treatment; five survived and one died. The other two expired because they did not receive appropriate antibiotic treatment. Although third-generation cephalosporin-resistant GNBM is rare, its incidence has been increasing over the past 5 years. In patients with nosocomially-acquired postneurosurgical GNBM, the presence of third-generation cephalosporin resistance should therefore be highly suspected. The appropriate choice of antibiotic is essential for their survival. PMID- 10378135 TI - Prevalence of anti-hepatitis A antibodies, hepatitis B viral markers, and anti hepatitis C antibodies among immigrants from the former USSR who arrived in Israel during 1990-1991. AB - The goal of this study was to assess the susceptibility of the sub-population of over 500,000 immigrants from the former USSR who came to Israel during 1989-94 to HAV infection, and to provide military physicians with estimates of the prevalence of HBV and HCV carriage in this sub-population. 987 males aged 17-49 and 195 females aged 17-19, reporting to military recruitment offices between December 1991 and March 1992 were tested. Anti-HAV, anti-HBV antibodies and hepatitis B surface antigen (HBsAg) were detected by using standard enzyme immunoassay (EIA) tests, and anti-HCV antibodies by a second-generation EIA and confirmed by a third-generation INNO-LIA test. It was found that in the 17-19 year age-group the prevalence of anti-HAV antibodies was 37%, anti-HBV was 12.8%, HBsAg was 3.0% and anti-HCV 1.3%. All markers were higher among males. The prevalence of anti-HAV and anti-HBs antibodies increased with age among males. That of HBsAg and anti-HCV antibodies increased with age overall. In the multiple logistic regression analysis, HAV and HBV seropositivity were significantly associated with the mother's education and republic of origin. It was concluded that the prevalence of anti-HAV antibodies is similar to that among the local population, which should not be considered at a higher risk of infection during military service. On the other hand, the higher prevalence of HBsAg and anti-HCV antibodies in this sub-population should heighten the awareness of the possibility of chronic liver pathology. PMID- 10378136 TI - Levels of nitric oxide, gamma interferon and interleukin-12 in AIDS patients with toxoplasmic encephalitis. AB - The production of nitric oxide (NO) by macrophages is important for the killing of intracellular pathogens, such as Toxoplasma gondii. Gamma interferon (IFN gamma) and lipopolysaccharide stimulate NO production. The aim of this study was to investigate the importance of NO, IFN-gamma and interleukin-12 (IL-12) in the host immune response in AIDS patients suffering from toxoplasmic encephalitis (TE). It was demonstrated that the production of NO, detected as nitrite/nitrate in the sera and in the cerebrospinal fluid (CSF) of 32 AIDS patients with TE, was normal. In addition, levels of IFN-gamma in the sera and in the CSF of patients with TE were not increased. In contrast, serum levels of IL-12 in these patients were significantly increased (6.5 +/- 7.1 pg/ml; P = 0.0368), compared to the control patients (1.7 +/- 3.5 pg/ml). Furthermore, increased but not significant levels of IL-12 were also observed in the CSF of patients with TE (2.2 +/- 4.7 pg/ml; controls: 0.5 +/- 1.9 pg/ml). The results of this study indicate that reactivation or recurrence of T. gondii infection in HIV-1-infected patients is probably due to a down-regulation of IFN-gamma along with a resulting non-optimal NO activity. PMID- 10378137 TI - Hepatitis E in Damascus, Syria. AB - A hospital-based study of acute hepatitis was conducted in Damascus, Syria, from 1995 to 1998. One hundred ninety-three sera from defined acute hepatitis cases were screened by ELISA for IgM anti-HAV, HBsAg, IgM anti-HBc and anti-HCV. Serum samples negative for all markers indicating recent infection by hepatitis A, B, or C were tested for HEV markers. Overall, 47 cases (24.4%) had no detectable hepatitis markers (non-A-E). HAV infection was detected in 71.2% of all viral hepatitis cases. Acute hepatitis B and C constituted 24 and 1.4% of the cases, respectively. Only five cases of acute hepatitis E were noted. Of 47 patients who had non-A-E hepatitis, fifteen (31.9%) tested positive for IgG anti HEV. This study provides indirect evidence that HEV is very likely to be endemic in Damascus, Syria. It reports for the first time the occurrence of hepatitis E in the country, a health problem that should be investigated further. PMID- 10378138 TI - Spontaneous non-typhoidal Salmonella peritonitis in patients with serious underlying disorders. AB - Non-typhoidal Salmonella is a rare cause of spontaneous bacterial peritonitis (SBP). Non-typhoidal Salmonella SBP has been reported in patients with relatively normal ascitic fluid protein levels. Five patients with non-typhoidal Salmonella SBP and a review of the literature are reported. These patients had chronic underlying disorders, such as malignancy, or other conditions causing immunosuppression. In previous reports, an ascitic fluid protein level above 1.5 g/dl was present in six patients, and under 1.5 g/dl in two. In the present report, ascitic fluid protein is above 2.5 g/dl in three patients and under 1.5 g/dl in one. Immunosuppression and the virulence of the organism seem to play a major role in non-typhoidal Salmonella SBP. Physicians should be alert to the possibility of non-typhoidal Salmonella infection in patients with SBP and normal protein levels in ascitic fluid. PMID- 10378139 TI - Cytomegalovirus-associated transverse myelitis in a non-immunocompromised patient. AB - Cytomegalovirus (CMV)-associated transverse myelitis is rare in immunocompetent patients. The case of a 54-year-old man is reported here who developed acute transverse myelitis with cerebrospinal fluid (CSF) alterations, suggesting a central nervous system infection. CMV-IgM positivity in serum and CMV isolated from blood, positive CMV PCR and positivity for pp65 antigen in blood, without viral antigens in the CSF and a positive response to therapy with ganciclovir (followed by progressive improvement) supported the diagnosis. PMID- 10378140 TI - Epstein-Barr virus and cytomegalovirus infections cause false-positive results in IgM two-test protocol for early Lyme borreliosis. PMID- 10378142 TI - Cephalometric study of the Apert syndrome in adolescence and adulthood. AB - This paper reports a cephalometric analysis of the craniofacial morphology in adolescents and adults with Apert syndrome. The sample comprised 26 patients with Apert syndrome (15 males and 11 females). The control group consisted of 153 adults (102 males and 51 females). Both lateral and frontal cephalograms were studied. The data were presented as mean plots of the craniofacial region together with data on some of the most significant findings. Marked differences were found in nearly all craniofacial regions except the mandible. The calvaria was increased in height and width but length was decreased. The cranial base showed marked protrusion of the greater wing of the sphenoid, which contributed to severe reduction of orbital volume and protrusion of the eyeglobe. Orbital volume was further reduced by maxillary hypoplasia in all three planes of space together with retrognathia. Maxillary height was extremely short and so was the nose. The width of the nasal cavity, height and depth of the bony nasopharynx, and the nasopharyngeal airway were all markedly reduced in size. The mandible was of fairly normal size and shape but was posteriorly inclined. Head posture was extended in relation to the cervical column. Total facial height was increased, whereas upper facial height was markedly reduced. Incisor occlusion showed mandibular overjet and open bite. Apert syndrome patients were then compared to a group of Crouzon syndrome patients. Marked and significant differences were found between the two syndromes in nearly all craniofacial regions, and craniofacial dysmorphology was generally more severe in Apert syndrome patients. PMID- 10378141 TI - Oral nitazoxanide and paromomycin inhalation for systemic cryptosporidiosis in a patient with AIDS. PMID- 10378143 TI - Nasal fossa malformations and paramedian facial cleft: new perspectives. AB - Choanal atresia may be associated with other cranio-facial malformations, including various degrees of nasal fossa malformation, and may be a part of paramedian facial clefts (as described by Tessier et al. [1977]). We identified five such cases with combined clinical elements corresponding to Tessier's paramedian facial cleft, including eyelid coloboma, mild to severe choanal and nasal fossa anomalies, ethmoidal hypoplasia and anterior skull base malformation, sometimes with proboscis lateralis and half-nose hypoplasia. These observations incited us, first, to elaborate a conception which accounts for the likely embryological mechanisms involved; second, to propose a new classification based on anatomical and pathogenic embryological considerations; and last, to propose the use of transpalatal approach to restore choanal permeability, since endonasal laser therapy is particularly dangerous in such cases. PMID- 10378144 TI - The effect of growth hormone therapy on craniofacial growth and dental maturity in children with Down syndrome. AB - Craniofacial growth was evaluated 3 years after termination of growth hormone (GH) therapy in ten Down syndrome (DS) children. The control group consisted of 16 age-matched children with DS. The treatment started at 6-9 months of age, and the duration was 36 months. There were no statistically significant differences in craniofacial development between DS children treated with GH or DS children not treated. In conclusion, the results of this study indicate that GH therapy for 36 months in children with DS did not change the craniofacial morphology compared to a group of DS children not given GH. PMID- 10378145 TI - Cytochemical identification of HSP110 during early mouse facial development. AB - Apoptotic cell death constitutes a common phenomenon observed during development. This process plays an important role in the regulation of cell populations and in early differentiation of embryonic organs. Several teratologic situations are considered as resulting in a dramatic increase of the apoptotic process. In mammalian cells, heat shock proteins (HSPs), expressed or increased in response to various stresses, act as molecular chaperones in physiological conditions. In order to determine specific histochemical markers of apoptotic cells in normal craniofacial development, we observed the expression of stress proteins (HSPs) 70, 86, and 110. The apoptotic pattern of mesectodermal cell death areas was confirmed using both nuclear staining (Feulgen) and specific labeling of DNA fragmentation (TUNEL). These areas are localized in the proximal parts of the first and second visceral arches. They are located in mesectodermal and ganglionic cells. Apoptotic mesectodermal populations strongly express HSP110, as shown by the cytochemical identification of HSP110 and by double staining HSP110 TUNEL, suggesting that this protein could be considered as a new marker for apoptotic embryonic cells, and could be used in further teratologic studies to better quantify induced cell death. PMID- 10378146 TI - Ectodermal ablation of the third branchial arch in chick embryos and the morphogenesis of the parathyroid III gland. AB - The parathyroid glands have been classically considered to be derivatives of the third and fourth pharyngeal pouches in most species, including humans. Furthermore, the presence of neural crest-derived cells in the parathyroid glands connective tissue has been apparently established. However, our previous studies have provided a new hypothesis on the origin of these glands in human and chick embryos. To determine the origin of the parathyroid III (P3) gland, ectoderm of the third branchial arch was cauterized in chick embryos at Hamburger and Hamilton's stage 19 (embryonic day 3). Cauterization of the ventral half of the ectoderm was followed by the non-formation, on the same side, of the P3 gland. When the dorsal half of the ectoderm was cauterized, both the right and left P3 glands formed. Our observations suggest that the ectoderm of the ventral half of the third branchial arch is necessary for the organization of the P3 gland. PMID- 10378147 TI - High levels of GM1-ganglioside beta-galactosidase in the salivary glands and GM1 like-ganglioside storage in parotids of deficient mice. AB - We have previously demonstrated high levels of GM1-ganglioside beta-galactosidase (beta-gal) in the salivary glands of Swiss-Webster mice (Nowroozi et al., J Craniofac Genet Dev Biol 18:51, 1998), and suggested that this activity reflects an important role for the lysosome in catabolism of salivary glycoconjugates. Here, we characterized and compared activities of lysosomal glycosidases among the salivary glands, spleen, and muscle of C57BL/6 mice, beta-gal hexosaminidase, and beta-glucuronidase activities are high in all three glands relative to muscle. Enzyme activities in the sublingual gland were substantially higher than in the submandibular and parotid glands. Spleen displays levels of activity that are comparable or higher (for beta-glucuronidase) than those in the salivary glands, whereas muscle displays substantially lower levels of these lysosomal glycosidases. In order to investigate the role of beta-gal in the salivary glands, we further characterized the salivary phenotype of knock-out mice deficient in this enzyme, mimicking human GM1-gangliosidosis. In contrast with the relative levels of beta-gal specific-activity among the salivary glands, only the parotid developed severe, generalized, degenerative histopathological changes in beta-gal-deficient knock-out mice. GM1-like-ganglioside, typically found at high levels only in the nerve tissue, where its exact function is still not clear, was demonstrated in storage vacuoles of the parotid glands of the deficient mice by binding of cholera toxin subunit B. Thus, beta-gal activity observed in the parotid gland most likely reflects its role in GM1-ganglioside catabolism, and this ganglioside, never previously reported in the salivary glands, may have a role in parotid exocrine secretory functions. beta-gal may also serve in secretory glycoprotein catabolism in other salivary glands, but this function may be non-essential for these glands. PMID- 10378148 TI - Morphological change of the nasopremaxillary suture in growing "toothless" osteopetrotic (op/op) mice. AB - Osteopetrotic (op/op) mice are known to commonly show a failure of tooth eruption. It is also well understood that masticatory function is highly associated with the craniofacial morphology of the growing mouse; however, the effects on sutural growth have not been studied. The present study was conducted to examine, in detail, the morphological and histological changes of the nasopremaxillary suture in these mutant mice and to assess a role of mechanical stress from mastication in the sutural growth. The width of the nasopremaxillary suture was measured on the section for the superior (P1), middle (P2), and inferior (P3) levels. The width of the nasopremaxillary suture for the P1 level in the normal mice fed a solid diet was significantly smaller in 30-day-old mice than in 15-day-old mice, whereas the width for the level P3 was significantly greater in the 30-day-old mice than in the 15-day-old mice. These changes in the sutural space were more prominent in the normal mice fed a solid diet than in the normal mice fed a granular diet. The sutural widths for all the levels became smaller in the 30-day-old op/op mice than in the 10-day-old op/op mice. The endocranial area of the nasopremaxillary suture showed synostosis in 30-day-old op/op mice. In both the normal and op/op mice, the number of tartrate-resistant acid phosphatase (TRAP)-positive cells was greatest at the age of 15 days. Moreover, the TRAP-positive cell number was smaller in the op/op mice than in the normal mice for all the experimental stages. Since, in general, mastication begins in mice after tooth eruption, i.e. from 15 to 30 days after birth, the present findings suggest that failure of tooth eruption and the reduced masticatory function restrict sutural modification. PMID- 10378149 TI - Novel electrode placements: time to reassess. PMID- 10378150 TI - Initial seizure threshold in the clinical practice of bilateral electroconvulsive therapy in Edinburgh, Scotland. AB - Initial seizure threshold was measured by empirical titration in 137 patients referred for bilateral electroconvulsive therapy (ECT) for the treatment of depressive illness. Treatment was given by an Ectron Series 5A ECT machine. The median and modal thresholds were 75 mC and the range was 50-200 mC. The only statistically significant relationship among initial threshold and age or gender was the correlation of age with threshold in men (rho = 0.46, p < 0.01). All patients (n = 22) younger than 30 years had an initial threshold < 100 mC, but age and gender did not predict threshold accurately for older men or women. PMID- 10378151 TI - Endpoint of ECT-induced elevation in heart rate. AB - Changes in the heart rate during electroconvulsive therapy (ECT) reflect seizure activity at a deeper brain site than shown by electroencephalography and motoric activity. Accordingly, such changes may provide additional information that is helpful in the evaluation of treatment quality. One basic measurement of heart rate change is the duration of ECT-induced tachycardia. In a prospective study, the electrocardiographic ECT records of 24 patients were rated for the abruptness of the endpoint of the seizure-induced elevation in heart rate; 19 showed abrupt endpoints and 5 showed gradual endpoints. The baseline heart rate of patients with abrupt endpoints (88 SD [standard deviation] 10 beats/min) was significantly lower (p = 0.00001) than those with gradual endpoints (118 SD 12 beats/min). A threshold occurred at a baseline heart rate of 100 beats/min, with abrupt endpoints below and gradual endpoints above. The data suggest that patients with low baseline heart rates might be likely to show bradyarrhythmia during the treatment, and corresponding precautions might be considered. PMID- 10378152 TI - Short-term effect of combined ECT and neuroleptic therapy in treatment-resistant schizophrenia. AB - Treatment-resistant schizophrenia (TRS) is a critical public health concern. Short-term treatment with electroconvulsive therapy (ECT), combined with neuroleptics, may increase the response rate in patients with TRS, when compared with either treatment alone. We conducted an open-trial study in 59 patients with TRS with acute exacerbations, by using bilateral ECT combined with flupenthixol (dose range, 12-24 mg/day). After the first sign of clinical improvement, all patients had to pass a 3-week stabilization period during which their clinical improvement had to be sustained. The patients had to receive at least 20 ECT treatments before being considered unresponsive to ECT. Thirty-one patients were ECT responders by our criteria, 19 were non-responders, and nine were dropouts. The responder group had more male patients, paranoid type, of younger age, shorter duration of illness and duration of the current episode, less family history of schizophrenia, and higher pretreatment GAF scores. They received a lesser number of ECT treatments, a less electrical charge used, and lower doses of flupenthixol (p < 0.05). Both positive and negative symptoms improved (p < 0.05), but positive symptoms responded to a greater extent. This study supports the therapeutic efficacy of combined treatment with ECT and neuroleptic drugs. A consensus in the definition of TRS is urgently required. PMID- 10378153 TI - Electroconvulsive therapy and mental retardation. AB - Reports on use of electroconvulsive therapy (ECT) in persons with mental retardation (MR) and mental illness are meager. We describe the successful use of ECT in the management of medication-resistant mental illness in persons with mental retardation. Details of the treatment of five patients with MR and psychotic disorders are reported. ECT was successful after the failure of adequate trials of traditional pharmacotherapy. Persons with MR did not have disproportionate adverse effects of ECT as a consequence of the MR. Four of the five cases reported received maintenance ECT. ECT should be considered in the affective and psychotic disorders occurring in persons with MR when traditional pharmacotherapy fails. PMID- 10378154 TI - Maintenance ECT in mentally retarded, treatment-resistant schizophrenic patients. AB - Three treatment-resistant schizophrenic patients with mental retardation were successfully treated with maintenance ECT combined with neuroleptics over a period of 6-18 months. There were no adverse effects. This report illustrates the benefits of ECT in schizophrenic patients with mental retardation. PMID- 10378155 TI - Treatment of Meige's syndrome with ECT. AB - A 62-year-old woman with Meige's syndrome failed to respond to several pharmacologic interventions. Her dystonias improved significantly after treatment with bilateral electroconvulsive therapy (ECT). However, the effect was not durable, lasting < or = 72 h. ECT is an effective treatment for many movement disorders including dystonias of differing etiologies. Its efficacy for Meige's syndrome is questionable. PMID- 10378156 TI - Electroconvulsive therapy for the treatment of neuroleptic malignant syndrome with psychotic symptoms: a report of five cases. AB - We report five cases of neuroleptic malignant syndrome (NMS) with psychotic symptoms treated with electroconvulsive therapy (ECT). Clinical response was observed after the first or the second session of ECT in every case, and the symptoms of NMS resolved by the third or fourth session. The mean time from the initial ECT to complete resolution was 6.0 days. No side effects from ECT were observed. Although the first treatment for NMS is pharmacotherapy using drugs such as dantrolene and bromocriptine, our results suggest that ECT is a useful therapeutic method for patients with NMS and psychotic symptoms. PMID- 10378157 TI - A simplified herbal formulation attenuates electroconvulsive shock-induced anterograde amnesia. PMID- 10378158 TI - Increment in reminiscing after ECT: possible connections to neuropsychologic changes. PMID- 10378159 TI - Induction of a reversible state of hypomania by rapid-rate transcranial magnetic stimulation over the left prefrontal lobe. PMID- 10378160 TI - Negative-pressure pulmonary edema: a potential hazard in patients undergoing ECT. PMID- 10378161 TI - Supraorbital and d'Elia electrode positions in unilateral ECT? PMID- 10378162 TI - Childhood abuse and sexual revictimization in a female Navy recruit sample. AB - To examine effects of childhood abuse on adult rape, 1,887 female Navy recruits were surveyed. Overall 35% of recruits had been raped and 57% had experienced childhood physical abuse (CPA) and/or childhood sexual abuse (CSA). Controlling for CPA, rape was significantly (4.8 times) more likely among women who had experienced CSA than among women who had not. In contrast, CPA (controlling for CSA) was unrelated to likelihood of adult rape. Alcohol problems and number of sex partners were examined as mediators. Although both variables predicted rape, their effects were independent of the effects of CSA. Finally, despite ethnic group differences in the prevalence of victimization, the predictors of rape did not differ significantly across ethnic groups. PMID- 10378163 TI - Exposure to violence and posttraumatic stress symptomatology among abortion clinic workers. AB - The intent of this study was to examine the relationship between exposure to abortion clinic violence, either as a victim or witness, and the reporting of posttraumatic stress disorder (PTSD) symptoms among clinic employees. Semi structured interviews with 71 clinic workers from eight abortion clinics in a Southeastern state were used for analyses. Findings showed that as victims, clinic workers experienced moderate forms of violence and witnessed greater variety and numbers of violent acts. Twenty-one percent of the sampled workers reported symptoms similar to the syndrome described in the DSM-IIIR/DSM-IV classification for PTSD. A multivariate analysis showed that even when controlling for significant life circumstances and stressors outside the clinic setting, witnessing violence was a significant predictor of PTSD symptomatology. PMID- 10378164 TI - Use of medical and mental health care by World War II survivors in The Netherlands. AB - This study examined the mental and medical health care utilization of World War II (WW II) survivors and the characteristics of survivors seeking professional health care. Forty seven years after the end of WW II, a random sample of 4,057 Dutch WW II survivors answered a four-page questionnaire; 1,461 persons subsequently answered an extensive follow-up questionnaire. Twenty-two percent had sought some form of health care for war-related complaints at some time since WW II. Most consultations were made in the 1940s. More consultations were made to general practitioners or to medical specialists as opposed to mental health specialists. Although the level of posttraumatic stress disorder (PTSD) symptoms was most important for discriminating between help-seeking and non-help-seeking respondents, 59% of the highly-exposed respondents with PTSD had not sought professional help in the years 1990-1992. The results show the importance of primary health care in recognizing PTSD symptoms and referring survivors to the appropriate professional helper. PMID- 10378165 TI - Virtual reality exposure therapy for PTSD Vietnam Veterans: a case study. AB - Virtual reality (VR) integrates real-time computer graphics, body tracking devices, visual displays, and other sensory input devices to immerse a participant in a computer-generated virtual environment that changes in a natural way with head and body motion. VR exposure (VRE) is proposed as an alternative to typical imaginal exposure treatment for Vietnam combat veterans with posttraumatic stress disorder (PTSD). This report presents the results of the first Vietnam combat veteran with PTSD to have been treated with VRE. The patient was exposed to two virtual environments, a virtual Huey helicopter flying over a virtual Vietnam and a clearing surrounded by jungle. The patient experienced a 34% decrease on clinician-rated PTSD and a 45% decrease on self-rated PTSD. Treatment gains were maintained at 6-month follow-up. PMID- 10378166 TI - PTSD among bereaved parents following the violent deaths of their 12- to 28-year old children: a longitudinal prospective analysis. AB - This study examined the prevalence of posttraumatic stress disorder (PTSD) among parents bereaved by the violent deaths of their 12- to 28-year-old children. A community-based sample of 171 bereaved mothers and 90 fathers was recruited by a review of Medical Examiner records and followed for 2 years. Four important findings emerged: Both parents' gender and children's causes of death significantly affected the prevalence of PTSD symptoms. Twice as many mothers and fathers whose children were murdered met PTSD caseness (full diagnostic) criteria compared with accident and suicide bereavement. Symptoms in the reexperiencing domain were the most commonly reported. PTSD symptoms persisted over time, with 21% of the mothers and 14% of the fathers who provided longitudinal data still meeting caseness criteria 2 years after the deaths. Parents who met caseness criteria for PTSD, compared with those who did not, were significantly different on multiple study variables. Both theoretical and clinical implications for the findings are discussed. PMID- 10378167 TI - Coping responses and posttraumatic stress symptomatology in urban fire service personnel. AB - Emergency workers, including urban fire fighters and paramedics, must cope with a variety of duty-related stressors including traumatic incident exposures. Little is known about coping responses of emergency workers or whether their coping responses predict future mental health outcomes. The previously formulated Coping Responses of Rescue Workers Inventory (CRRWI) underwent a principal components analysis employing a sample (N = 220) of urban fire fighters and paramedics. Six empirically and theoretically distinct CRRWI components were identified which were relatively stable over a 6-month period. Scores on one of the CRRWI scales, but neither years of service nor their past half year's traumatic incident exposures, predicted future changes in self-reports of posttraumatic stress symptomatology. PMID- 10378168 TI - Compelled attention: the effects of viewing trauma-related stimuli on concurrent task performance in posttraumatic stress disorder. AB - We examined the ability of Vietnam veterans with PTSD to focus attention on a primary digit detection task while concurrently viewing neutral or Vietnam related picture and word distractors. Controlling for combat exposure, military service, and psychopathology, veterans with PTSD took longer to detect the target when Vietnam-related pictures were distractors. There were no reaction time differences when word stimuli were distractors. The latency effect was specific to trials with trauma-related pictures and did not spread to neutral trials interleaved within a mixed block of trauma and neutral pictures. Individuals with PTSD recalled proportionally more Vietnam-related words than other groups, implying differential attention to Vietnam-related words. Attending to trauma related pictures interferes with performance of a concurrent task by individuals with PTSD. PMID- 10378169 TI - Partners' ratings of combat veterans' PTSD symptomatology. AB - The current study examined the concordance of combat veterans' scores on the Mississippi Scale for Combat-Related PTSD, with scores on a parallel version of that instrument completed by partners to assess veterans' symptoms. Further, the study examined the impact of quality of the marital relationship on score concordance. Bivariate and multiple regression were used with a sample of 466 veteran-partner dyads obtained from the National Vietnam Veterans Readjustment Study. There was moderate agreement in symptom reporting between veterans and their partners and little evidence to suggest that the quality of the relationship impacted upon the association between partner and veteran scores. PMID- 10378170 TI - A comparison of ICD10 and DSM-IV criteria for posttraumatic stress disorder. AB - The assumption that participants receiving an ICD10 diagnosis of posttraumatic stress disorder (PTSD) will also receive a DSM-IV diagnosis of PTSD was tested. Data were gathered for 1,364 participants using the Composite International Diagnostic Interview (CIDI). The 12-month prevalence of PTSD was 3% for DSM-IV and 7% for ICD10 Diagnostic Criteria for Research (ICD10-DCR). The agreement between the two systems was fair (kappa = .50). Forty eight percent of the discrepancies between the systems were accounted for by the additional criterion requiring clinically significant distress or impairment included in DSM-IV. The inclusion of symptoms of general numbing of responsiveness accounted for 18% of the discrepancies. It is concluded that ICD10-DCR PTSD cannot be assumed to be identical to DSM-IV PTSD. PMID- 10378171 TI - Effects of attribution of responsibility for motor vehicle accidents on severity of PTSD symptoms, ways of coping, and recovery over six months. AB - In light of Delahanty et al.'s (1997) identification of attribution of responsibility for a motor vehicle accident (MVA) as a powerful determinant of initial level of distress from the trauma and of early remission of PTSD, we reexamined data from Blanchard and Hickling's (1997) prospective follow-up of 158 MVA survivors. Despite differences between the two samples (Delahanty sample recruited from hospitals 2-3 weeks post-MVA and predominantly male; our sample recruited from outpatient care 1-4 months post-MVA and predominantly female) we replicated Delahanty's findings: those with PTSD who blame themselves for the MVA are less symptomatic initially and recover more rapidly in the first 6 months than those with PTSD who blame another party for the accident. PMID- 10378172 TI - Effects of mode of writing on emotional narratives. AB - The authors hypothesized that writing longhand about a stressful experience, compared to typing, arouses greater negative emotion. Eighty college students were randomly assigned to describe either a neutral or stressful topic by typing or writing longhand, in a 2 x 2 factorial design. Students describing the stressful topic, compared to the neutral topic, wrote for a longer period, used more words, and reported greater negative and less positive affect. Consistent with prediction, writing about a stressful experience longhand induced greater negative affect than typing, and led to more self-rated disclosure. These findings suggest a method whereby therapists can help patients control their levels of negative affect when producing a trauma narrative. PMID- 10378173 TI - Acute intrusive and avoidant PTSD symptoms as predictors of chronic PTSD following burn injury. AB - Several studies have endeavored to learn if acute PTSD symptoms are predictive of chronic PTSD, with equivocal results. In the present study, acute intrusive and avoidant PTSD symptoms were analyzed as possible predictors of chronic PTSD following burn injury. Results showed that baseline IES scores, within one week of injury, were significantly different for those who were later diagnosed with chronic PTSD. Additional analyses, undertaken to assess the relative importance of each symptom group in predicting chronic PTSD, indicated that both the presence, per se, and severity of acute avoidant symptoms predicted chronic PTSD. PMID- 10378174 TI - MMPI-2 data for Australian Vietnam Veterans with combat-related PTSD. AB - Considerable attention has been devoted to the MMPI in the assessment of combat related PTSD. To date, published data have focused almost exclusively on American Vietnam veterans. This study investigated MMPI-2 profiles of 100 Australian Vietnam veterans admitted to an intensive PTSD treatment program. Comparisons with United States (U.S.) data suggested strong similarities between the American and Australian populations in terms of F-scale elevations and typical 3-point code types (8-7-2). However, the American samples showed relatively higher elevations of Scales 4 and 6, suggesting social alienation and a tendency to externalize, while a subgroup of Australian veterans showed a greater propensity for somatization (Scale 1). The results provide overall support for the generalizability of American MMPI data to an alternative cultural group of combat veterans. PMID- 10378175 TI - Psychometric properties of a hurricane coping self-efficacy measure. AB - This brief report describes the psychometric properties of an instrument designed to measure Hurricane Coping Self-Efficacy (HCSE). Survivors of Hurricane Andrew (n = 165) and Hurricane Opal (n = 63) completed the HCSE and assessments of optimism, social support, distress, and resource loss. Principal components factor analyses revealed a unidimensional structure for the HCSE. Internal consistency of the HCSE was strong. In both samples, HCSE was positively associated with optimism and social support, but negatively associated with general psychological distress, trauma related distress, and resource loss. Finally, hierarchical regression analyses demonstrated that the HCSE explained a significant amount of experimental variance for intrusive thoughts and avoidance after controlling for social support, lost resources, and optimism. PMID- 10378176 TI - Quality of life and posttraumatic stress disorder: a pilot study assessing changes in SF-36 scores before and after treatment in a placebo-controlled trial of fluoxetine. AB - In this small pilot study, we evaluated quality of life for 16 posttraumatic stress disorder (PTSD) patients by administering the Medical Outcomes Study 36 item Short-Form Health Survey (SF-36) at baseline and endpoint during a 12-week double-blind trial of fluoxetine and placebo. At baseline, our subjects reported greater impairment relative to subjects with major depression or obsessive compulsive disorder on several SF-36 domains. Significant effects of fluoxetine relative to placebo were observed for vitality, social functioning, and mental health. Overall, PTSD was associated with greatly reduced quality of life, but considerable improvement was achieved through treatment. PMID- 10378177 TI - Divalproex in posttraumatic stress disorder: an open-label clinical trial. AB - Posttraumatic stress disorder (PTSD) is characterized by intrusive, avoidance, and hyperarousal symptoms. This study was conducted to investigate the effectiveness of divalproex in reducing PTSD symptoms, depression, and anxiety in patients with PTSD. Sixteen patients with a DSM-IV diagnosis of PTSD at the Albuquerque VAMC outpatient PTSD treatment program received an open-label trial of divalproex. The patients were evaluated at baseline and at 8 weeks by a trained rater using the Clinician Administered PTSD Scale (CAPS), the Hamilton Rating Scale for Depression (HAM-D) and the Hamilton Rating Scale for Anxiety (HAM-A). Plasma valproate levels were measured at the 8-week post-treatment assessment. Three patients stopped the medications due to side effects. Intrusion and hyperarousal symptoms decreased significantly, while no significant change was seen in avoidance/numbing symptoms. Depressive symptoms, as measured by the HAM-D, unexpectedly decreased at post-treatment assessment. HAM-A scores also decreased significantly. Controlled trials are needed to further study the efficacy of divalproex in the treatment of PTSD. PMID- 10378178 TI - The subdivisions of the guinea pig vestibular complex revealed by acetylcholinesterase staining. AB - A detailed map of the vestibular nuclear complex of the guinea pig has been established by Gstoettner and Burian (1987), using cytoarchitectonic (cresyl violet staining) and fiberarchitectonic criteria. However, the exact borders between the different subdivisions are not always evident in Nissl stained sections. In the present study, serial sections of the vestibular nuclei of the guinea pig were stained to visualize acetylcholinesterase (AChE) activity, and compared with corresponding sections stained with cresyl violet. All of the subdivisions of the vestibular nuclear complex previously described are more readily distinguished in AChE than in Nissl preparations. The AChE reactivity also shows that the medial vestibular nucleus extends more rostrally than previously described. Furthermore, it questions whether the area classically referred to as the rostral pole of the descending vestibular nucleus belongs to the descending vestibular nucleus or to the lateral vestibular nucleus (LV). Finally, a morphometric analysis performed on cresyl violet stained sections shows that (1) in the caudal LV, the neurons of the ventromedial extension are smaller than those of the dorsolateral extension and that (2) in the rostral LV, the ventromedial division contains a larger ratio of smaller neurons than the dorsolateral one. PMID- 10378179 TI - Importance of the vestibular system in visually induced nausea and self-vection. AB - The objective of this study was to determine the importance, if any, of the non auditory labyrinth of the inner ear in visually induced nausea and self-vection in subjects exposed to a moving visual field with and without concomitant pitching head movements. Subjects treated were 15 normals, 18 unilateral labyrinthectomies and 6 bilateral labyrinthectomies. The findings show a higher incidence of pseudo-Coriolis induced nausea in normal subjects compared to unilateral and bilateral labyrinthectomized subjects. When the subjects were exposed to the moving visual field only (no head movement), pronounced self vection occurred in all subjects, but with earlier onset in the bilateral labyrinthine defective subjects as compared to normal and unilateral defective subjects. The subjective intensities of self-vections reported by labyrinth defectives were much more pronounced as compared to normal subjects, and it is apparent that visual input in these subjects achieves much more importance in maintaining compensatory eye movements, and the gain of neck reflexes is enhanced. The findings that visual stimulation is more effective in producing the disabling effects after labyrinthine destruction could possibly be explained by enhancement of vision after loss of labyrinthine sensory input, and the gain in neck reflexes is also enhanced after labyrinthectomy. PMID- 10378180 TI - "Torso Rotation" experiments. 4: the role of vision and the cervico-ocular reflex in compensation for a deficient VOR. AB - Acute, reversible changes in human vestibular function can be produced by exposure to "Torso Rotation" (TR), a method involving the overuse of certain types of simple, self-generated movements. A single session results in multiple, short-lasting aftereffects, including perceptual illusions, VOR gain reduction, gaze and postural instability, and motion sickness. With repeated exposure, motion sickness susceptibility disappears and gaze stability improves. VOR gain continues to be reduced, however. Therefore, another gaze stabilizing system must come into play. Are visual and/or neck inputs involved in this functional compensation? Six subjects participated in this 7-day experiment. Eye and head movements were measured during 2 tests: 1) voluntary "head only" shaking between 0.3 and 3.0 Hz (lights off) and 2) voluntary "head and torso" shaking, moving the upper body en bloc (neck immobilized). Measurements were obtained before and repeatedly after TR. Velocity gain (eye velocity/head velocity) was determined for each of these tests. Each day, mean velocity gain during "head only" shaking in the dark (averaged over 1.0 to 2.0 Hz) dropped significantly after TR (P < 0.01), with no long-term improvement (P > 0.9). Similar results, although more noisy, were obtained for "head and torso" shaking. As a control, EOG calibration data confirmed that gaze stability in the light did improve over the 7 days of testing. This experiment demonstrates that the reduction in gaze instability following repeated exposure to TR results from an increased use of vision. It excludes the VOR, the COR, and predictive mechanisms (including efference copy) as contributors. In addition, in the 20 minutes following TR completion, gaze stability recovered less than during previous VOR testing in the dark. These results are compatible with the motion that exposure to TR leads to a change in sensorimotor strategy involving a de-emphasis of vestibular inputs. PMID- 10378181 TI - Postural sway with earth-fixed and body-referenced finger contact in young and older adults. AB - Sensory information from lightly touching a reference with the hand is known to influence postural sway in young adults. The primary aim of this study was to compare the influence of finger contact (FC) with an earth-fixed reference to the influence of FC with a body-fixed reference. A second goal of this study was to determine if FC is used differently by older adults compared to younger adults. Using a force plate, center of pressure at the feet was recorded from blindfolded young and older subjects during several conditions. Subjects either did or did not lightly touch a force-sensitive plate that was either earth-fixed or moved forward and backward in synchrony with body sway (that is, sway-referenced). In addition, support surface conditions were also varied, including a fixed floor and a sway-referenced floor using an Equitest. Results showed that the type of FC, floor condition, and age each had an effect on postural sway. Touching an earth-fixed plate decreased postural sway as compared to no touching, while touching a sway-referenced plate increased sway. This influence of FC was enhanced when the floor was sway-referenced. Although older subjects swayed more than young subjects overall, no age-FC interactions occurred, indicating that FC was not utilized differently between the age groups. This study suggests that FC cannot be disregarded as erroneous, especially when proprioceptive information from the legs is distorted. Further, FC is integrated with other sensory information by the postural control system similarly in young and older persons. PMID- 10378182 TI - Evidence that the ginkgo biloba extract, EGb 761, neither accelerates nor enhances the rapid compensation of the static symptoms of unilateral vestibular deafferentation in guinea pig. AB - The concentrated Ginkgo biloba extract, EGb 761, has previously been reported to enhance and accelerate vestibular compensation following unilateral vestibular deafferentation (UVD), in particular, compensation of the dynamic postural symptoms such as locomotor dysequilibrium. However, many of these studies have not included a complete analysis of the static symptoms of UVD, such as spontaneous nystagmus (SN), yaw head tilt (YHT), and roll head tilt (RHT), nor have they included a dose-response analysis or vehicle controls for EGb 761. The aim of the present study was to examine the effects of the EGb 761 extract on static vestibular compensation in guinea pig, using a dose-response analysis and both vehicle and saline controls. Analysis of variance showed that there was a significant decrease in SN frequency (P < 0.05) and a significant change in the rate of SN compensation (P < 0.05), using 3 i.p. injections of EGb 761 (25, 50, or 100 mg/kg), or vehicle, or saline, at 0, 25, and 40 h post-UVD. However, post hoc testing revealed that this was due entirely to significant differences between the saline and vehicle groups at 35, 40, and 50 h post-UVD (P < 0.05 in all cases) and between the saline and the 100 mg/kg and 25 mg/kg EGb 761 groups at 35 and 50 h post-UVD, respectively (P < 0.05 for both comparisons); there were no significant differences between the vehicle and drug groups at any time. YHT and RHT were not significantly different between the drug, saline, and vehicle groups. In a second set of experiments, the 50 and 100 mg/kg EGb 761 i.p. injection frequencies were doubled. However, once again, neither SN nor YHT were significantly different between the EGb 761 groups and the vehicle controls. These results suggest that 1) EGb 761 does not significantly enhance or accelerate compensation of the static symptoms of UVD in guinea pig and 2) the EGb 761 vehicle may exert some effects on its own. Therefore, EGb 761 may be of limited use in the treatment of acute vestibular dysfunction in humans. PMID- 10378183 TI - Comparison of two head autorotation tests. AB - The head autorotation test is a novel method for studying the high-frequency vestibuloocular reflex without heavy machinery to generate whole-body rotation. Despite many studies with the test, the method is far from standardized, and no comparison has been made of different versions of the test. The objective of this study was to compare the vestibuloocular reflex of 100 healthy subjects measured simultaneously with two versions of the head autorotation test. Gain, phase, asymmetry, and the frequency bands reached were determined in the frequency bands of 1, 2, 3, 4, and 5 Hz. The gain measured with both tests was close to unity (range 0.95-1.04) from 1 to 4 Hz and about 0.9 at 5 Hz. In the test developed by Vorteq the phase lagged (-7 to -21 degrees) in all the frequency bands, and it differed significantly from the phase lead of 2 to 5 degrees that was measured by the other test. The asymmetry measured with the Vorteq test increased continuously from 1.5% (1 Hz) to 5.7% (5 Hz). The results of the tests showed intersubject variation, which was larger in the higher frequency bands. In conclusion, the high-frequency vestibuloocular reflex of healthy subjects can be quantified with active head oscillation. Both tests produced similar gain results, but the phase results differed systematically. Thus, the results of different head autorotation tests may not be directly comparable. PMID- 10378184 TI - The effect of d-amphetamine on optokinetic nystagmus in the guinea pig. AB - The effect of d-amphetamine oral administration in doses of 1-2.5 mg/kg on horizontal optokinetic nystagmus (OKN) and after nystagmus (OKAN) was investigated in the guinea pig. Eye movements were recorded by means of the electromagnetic search-coil technique. After amphetamine administration the range of stimulus velocities effective for eliciting OKN was 10-20 deg/s higher than before treatment. The mean values and the fluctuations of the eye velocity during slow nystagmus phases before and after treatment did not differ. Administration of amphetamine led to 2-8 s increase in OKAN duration. The OKAN prolongation did not depend on stimulation velocity. The dependency of OKAN duration on stimulation velocity was well approximated by a linear regression. The slope of the regression line was 0.160 +/- 0.022 before and 0.177 +/- 0.028 after treatment. Similarity in the coefficients indicates that amphetamine did not alter the relationship between the velocity of optokinetic stimulus and the duration of after nystagmus. Constant prolongation of OKAN over the whole range of stimulation velocities could reflect a constant shift in activity of neurons representing the velocity storage. The effects observed on OKN gain curves and the increase in OKAN duration did not display a clear dependency on the dosages of d-amphetamine used in the experiments. We assume that the effects of treatment reflected a general increase in attentiveness and motility of animals resulting from the arousal action of amphetamine. PMID- 10378185 TI - Recovery of vestibulo-ocular reflex-function in subjects with an acute unilateral peripheral vestibular deficit. AB - The centrally controlled compensation for a reduced horizontal vestibulo-ocular reflex (VOR) gain caused by a unilateral afferent deficit is usually studied following a selective surgical procedure which completely lesions the vestibular nerve or blocks the horizontal semicircular canal. The more common, unilateral, vestibular deficit encountered clinically, is a partial loss of peripheral vestibular function, following which peripheral recovery and/or central compensation may occur. We investigated changes of the VOR gain in response to a sudden, idiopathic, unilateral vestibular deficit in 64 subjects by examining the responses to low-frequency, whole-body, rotations about an earth vertical axis with different accelerations (5, 20 and 40 deg/sec2) during in- and out-patient visits separated by 4 months in an attempt to identify changes brought about by peripheral recovery and by central compensation processes. Peripheral function was assumed to be measured by the response to caloric irrigation. It improved some 30% on average between the two visits. VOR responses for rotations towards the deficit side also improved between the two visits. Most improvement occurred for 20 deg/sec2 accelerations. However, the correlation coefficient between rotation and caloric responses was always less than 0.6. Unlike caloric responses which improved over time, responses for rotations to the intact side did not change between the visits. For this reason, the majority of observed VOR rotation responses were nearly symmetrical at the time of the second visit, despite being below normal levels. These findings suggest that both peripheral recovery and central compensation processes help restore symmetrical VOR function for head rotations after a partial unilateral vestibular deficit. However the improvement of VOR response symmetry, particularly to slow (< 40 deg/sec2) accelerations, is largely independent of the recovery of peripheral sensitivity. PMID- 10378186 TI - Subjective visual vertical in peripheral unilateral vestibular diseases. AB - In humans, the perception of vertical is provided by input from various sensorineural organs and pathways: vision, eye-movements, and proprioceptive and vestibular cues, particularly from the otolithic organs and graviceptive pathways. Well known in several types of brainstem lesions, subjective visual vertical (SVV) abnormalities may also be observed after peripheral vestibular lesions, such as surgical deafferentation, with a deviation directed toward the operated ear. Subjective visual vertical abnormalities are presumably related to a lesion of the otolithic organs and/or to changes in the afferent graviceptive pathways. The goal of this prospective study was to measure the SVV and to define the influence of the otolithic organs in patients suffering from various types of peripheral vestibular diseases: unilateral sudden cochleo-vestibular loss, so called "viral labyrinthitis" (VL), sudden idiopathic unilateral peripheral vestibular loss, so-called "vestibular neuritis" (Ne). Data were compared with findings after unilateral surgical deafferentations such as vestibular neurectomy (VN) and labyrinthectomy (Lab). Subjective visual vertical was measured with a binocular test (vertical frame) and a monocular test (Maddox rod). In all patients, after VN and Lab, the SVV showed a 10-30 degrees tilt with the vertical frame (N: 0 +/- 2 degrees), 5-15 degrees with the Maddox rod (N: 0 +/- 4 degrees). With the vertical frame, SVV was tilted > 2 degrees in VL (47%) and in Ne (37%); the Maddox rod showed a > 4 degrees tilt in VL (41%) and in Ne (42%). The tilt was directed toward the affected ear. Our results demonstrate that SVV is frequently tilted in acute peripheral vestibulopathies such as VL and Ne. These findings suggest that otolithic function is implicated in the deficit depending on the extent and/or the localisation of the peripheral vestibular lesion. PMID- 10378187 TI - Solutions of the BCM learning rule in a network of lateral interacting nonlinear neurons. AB - We introduce a new method for obtaining the fixed points for neurons that follow the BCM learning rule. The new formalism, which is based on the objective function formulation, permits analysis of a laterally connected network of nonlinear neurons and allows explicit calculation of the fixed points under various network conditions. We show that the stable fixed points, in terms of the postsynaptic activity, are not altered by the lateral connectivity or nonlinearity. We show that the lateral connectivity alters the probability of attaining different states in a network of interacting neurons. We further show the exact alteration in presynaptic weights as a result of the neuronal nonlinearity. PMID- 10378188 TI - Visual coding and the phase structure of natural scenes. AB - Although it is now well known that natural images display consistent statistical properties which distinguish them from random luminance distributions, this ecological approach to vision has so far concentrated on those second-order image statistics which are quantified by image power spectra, and it appears to be the image phase spectra which carry the majority of the image-intrinsic information. The present work describes how conventional nth-order statistics can be modified so that they are sensitive to image phase structure only. The modified measures are applied to an ensemble of natural images, and the results show that natural images do have consistent higher-order statistical properties which distinguish them from random-phase images with the same power spectra. An interpretation of this finding in terms of higher-order spectra suggests that these consistent properties arise from the ubiquity of edge structures in natural images, and raises the possibility that the properties of ideal relative-phase-sensitive mechanisms could be determined directly from analyses of the higher-order structure of natural scenes. PMID- 10378189 TI - Processing images by semi-linear predictability minimization. AB - In the predictability minimization approach, input patterns are fed into a system consisting of adaptive, initially unstructured feature detectors. There are also adaptive predictors constantly trying to predict current feature detector outputs from other feature detector outputs. Simultaneously, however, the feature detectors try to become as unpredictable as possible, resulting in a co-evolution of predictors and feature detectors. This paper describes the implementation of a visual processing system trained by semi-linear predictability minimization, and presents many experiments that examine its response to artificial and real-world images. In particular, we observe that under a wide variety of conditions, predictability minimization results in the development of well-known visual feature detectors. PMID- 10378190 TI - Entropy optimization by the PFANN network: application to blind source separation. AB - The aim of this paper is to present a study of polynomial functional-link neural units that learn through an information-theoretic-based criterion. First the structure of the neuron is presented and the unsupervised learning theory is explained and discussed, with particular attention being paid to its probability density function and cumulative distribution function approximation capability. Then a neural network formed by such neurons (the polynomial functional-link artificial neural network, or PFANN) is shown to be able to separate out linearly mixed eterokurtic source signals, i.e. signals endowed with either positive or negative kurtoses. In order to compare the performance of the proposed blind separation technique with those exhibited by existing methods, the mixture of densities (MOD) approach of Xu et al, which is closely related to PFANN, is briefly recalled; then comparative numerical simulations performed on both synthetic and real-world signals and a complexity evaluation are illustrated. These results show that the PFANN approach gives similar performance with a noticeable reduction in computational effort. PMID- 10378191 TI - Visual segmentation by contextual influences via intra-cortical interactions in the primary visual cortex. AB - Stimuli outside classical receptive fields have been shown to exert a significant influence over the activities of neurons in the primary visual cortex. We propose that contextual influences are used for pre-attentive visual segmentation. The difference between contextual influences near and far from region boundaries makes neural activities near region boundaries higher than elsewhere, making boundaries more salient for perceptual pop-out. The cortex thus computes global region boundaries by detecting the breakdown of homogeneity or translation invariance in the input, using local intra-cortical interactions mediated by the horizontal connections. This proposal is implemented in a biologically based model of V1, and demonstrated using examples of texture segmentation and figure ground segregation. The model is also the first that performs texture or region segmentation in exactly the same neural circuit that solves the dual problem of the enhancement of contours, as is suggested by experimental observations. The computational framework in this model is simpler than previous approaches, making it implementable by V1 mechanisms, though higher-level visual mechanisms are needed to refine its output. However, it easily handles a class of segmentation problems that are known to be tricky. Its behaviour is compared with psycho physical and physiological data on segmentation, contour enhancement, contextual influences and other phenomena such as asymmetry in visual search. PMID- 10378192 TI - The Sixth John M. Kinney-Nestle Clinical Nutrition Award for Enteral Nutrition. Recombinant human tumor necrosis factor and recombinant murine interleukin-1 alter the binding of Escherichia coli to intestine, mucin glycoprotein, and the HT29-C1 intestinal cell line: a context. PMID- 10378193 TI - Cytokines, intestine, and bacteria. PMID- 10378194 TI - The Sixth John M. Kinney-Baxter Healthcare Award for Parenteral Nutrition. Water and sodium restriction during preoperative TPN in severely malnourished patients: the history. PMID- 10378195 TI - Preoperative parenteral nutrition--standard or special regime? PMID- 10378196 TI - The Sixth John M. Kinney International Awards for Nutrition and Metabolism . "Glutamine-enhanced bacterial killing by neutrophils from postoperative patients": background. PMID- 10378197 TI - The role of glutamine in modulating the body's immune function. PMID- 10378198 TI - Energy expenditure and substrate utilization in mechanically ventilated children. AB - The objective of the study was to determine the value of indirect calorimetry and nitrogen balance (N balance) in order to evaluate the current feeding protocols of mechanically ventilated children. The study was designed as a cross-sectional prospective study. In 36 mechanically ventilated children energy expenditure was measured by indirect calorimetry, and total urinary nitrogen excretion (TUN) was determined. Substrate utilization and respiratory quotient (RQ) were calculated from the measured values of oxygen consumption (VO2), carbon dioxide production (VCO2), and TUN. The RQ was compared with the RQ of the macronutrients administered (RQmacr) according to the modified criteria of Lusk. In results, the total measured energy expenditure (TMEE) showed a wide variation (range 155-272 kJ.kg-1.d-1). The N balance was positive in 20 and negative in 16 patients. The ratio of caloric intake/TMEE was significantly higher in patients with a positive N balance (1.50 +/- 0.06) as compared with those with a negative N balance (0.8 +/- 0.1, P < 0.001). There was a significant relationship between the difference of RQ-RQmacr versus the ratio caloric intake/TMEE (r = 0.72, P < 0.001). Carbohydrate and fat utilization were not significantly different in patients with a positive or negative N balance. Protein utilization was significantly higher in those patients with a negative N balance. We concluded that measurement of TMEE with indirect calorimetry results in accurate determination of energy needs in critically ill mechanically ventilated children. Feeding according to or in excess of the TMEE is correlated with a positive N balance. A combination of the RQ and the RQmacr can be helpful in differentiating under- or overfeeding. PMID- 10378199 TI - Plasma zinc, copper, and erythrocyte superoxide dismutase in children with phenylketonuria. AB - Children with phenylketonuria (PKU) are treated with semisynthetic diets restricted in phenylalanine (PHE). The formulae must supply those trace elements and vitamins that are usually supplied by whole protein foods. We studied the effects of phenylalaline restricted diets in 42 children with PKU (P) and 31 normal (N) children, aged 1-12 y, divided into two groups (below and above 7 y). Plasma zinc and copper were analyzed by means of atomic spectrophotometry, and superoxide dismutase (CuZnSOD) activity was measured in erythrocytes, through NBT inhibition and its profile, as determined by isoelectric focalization. Plasma zinc of PKU children > or = 7 years old was significantly lower than that in the control group (17 mumol/L versus 20 mumol/L) but still within the normal range; in children < 7 years no substantial differences were found between the two groups. Plasma copper was not statistically different between PKU and normal children. Qualitative activity of CuZnSOD presented the same electrophoretic profile in both normal and PKU. Quantitative activity was not different in both P (1210 U/g Hb < 7 versus 1328 U/g hemoglobin (Hb) > or = 7) and N (1675 U/g Hb < 7 versus 1367 U/g Hb > or = 7). We concluded that children with PKU presented normal mean levels of zinc and copper, with preserved function, measured by enzyme activity. PMID- 10378200 TI - Body weight changes with protease inhibitor treatment in undernourished HIV infected patients. AB - Effective reduction of HIV replication by protease inhibitor (PI) treatment was expected to reverse some of the weight loss associated with HIV infection. Body weight changes in undernourished HIV-infected patients starting PI treatment were compared to its virologic and immunologic effects. This was designed as a retrospective study using prospectively collected weight data; the setting was the HIV outpatient department of a university hospital. Among 223 consecutive HIV positive patients starting treatment with PI February 1996 to September 1997, 63 undernourished patients were evaluable. The main outcome measures were weight trend, calculated by linear regression of a patient's weight versus time, and its change from a 4-14-wk baseline period to the first 14 wk, and 28 wk, after treatment. In our results, weight trend remained unchanged (baseline, +0.4 +/- 4.0 kg/100 d; 14 wk, +0.7 +/- 4.1 kg/100 d, and 28 wk, +1.0 +/- 3.4 kg/100 d, n.s.). Reduction of viremia and increase in CD4 cell count were unrelated to weight trends. Treatment with PI did not result in an improved weight trend. Altered body composition with PI treatment, as observed in other studies, does not seem to result in body weight changes. Drug side effects may have counteracted any positive effects. The metabolic and nutritional impact of effective antiviral treatment merits further study. PMID- 10378201 TI - Development of a valid and reliable malnutrition screening tool for adult acute hospital patients. AB - Nutrition screening identifies individuals who are malnourished or at risk of becoming malnourished and who may benefit from nutrition support. The aim of this study was to develop a simple, reliable and valid malnutrition screening tool that could be used at hospital admission to identify adult acute patients at risk of malnutrition. The sample population included 408 patients admitted to an Australian hospital, excluding pediatric, maternity, and psychiatric patients. The ability of various nutrition screening questions to predict subjective global assessment (SGA) were examined in contingency tables. The combination of nutrition screening questions with the highest sensitivity and specificity at predicting SGA was termed the malnutrition screening tool (MST), and consisted of two questions regarding appetite and recent unintentional weight loss. Subjects who were at risk of malnutrition according to the MST had significantly lower mean values for the objective nutrition parameters (except immunologic parameters) and longer length of stays than subjects who were not at risk of malnutrition. Therefore convergent and predictive validity of the MST was established. The interrater reliability of the malnutrition screening tool was high (93-97%). The MST is a simple, quick, valid, and reliable tool which can be used to identify patients at risk of malnutrition. PMID- 10378202 TI - Effects of megestrol acetate and testosterone on body composition in castrated male Sprague-Dawley rats. AB - The interrelationships among sex hormones, caloric intake, and intermediary metabolism in health and disease are uncertain. Studies in malnourished patients with AIDS and cancer show that megestrol acetate (MA) therapy increases appetite, body weight, and body fat, while it decreases serum testosterone (T) concentration. In this study, the separate and combined effects of MA and T were investigated in 65 young adult, male, castrated, Sprague-Dawley rats who received subcutaneous implants containing placebo, MA, T, or both MA and T for 11 wk. By hierarchical multiple regression analysis, MA therapy decreased weight gain and food intake (P < 0.01), increased body fat (P = 0.024), decreased body protein (P < 0.001), and decreased the portion of calories accrued as protein rather than fat (P ratio, P < 0.03). T alone decreased fat (P < 0.03), but had no significant effect on food intake, the relative number of consumed calories utilized for growth (food efficiency), body weight, or protein. The interaction of MA and T did not affect food intake or food efficiency, but increased body weight (P < 0.02), protein (P < 0.05) and the P ratio (P < 0.02). The portion of weight gain as fat was reduced from 47.3% with MA alone to 27.4% when MA and T were combined. Thus, megestrol acetate has significant antianabolic effects that are independent of its effects upon food intake. The addition of testosterone to megestrol acetate partially antagonized MA's inhibition of lean mass accretion in these rats. PMID- 10378203 TI - Effects of alimentary intact proteins and their oligopeptide hydrolysate on growth, nitrogen retention, and small bowel adaptation in inflammatory turpentine rat. AB - The effects of dietary proteins given as whole proteins (WP) or as a peptide hydrolysate (PH) on growth, nitrogen retention, and small bowel adaptation were assessed using two groups of male Wistar rats. Measurements were made 18, 42, and 66 h after acute inflammation induced by subcutaneous injections of 0.125 mL turpentine and in two control groups (n = 12). The two diets had the same caloric, nitrogen, vitamin, and mineral content. The WP diet resulted in better weight gain, nitrogen retention, and small intestinal adaptation by control rats than did the PH diet. Loss of body weight after 18 h of acute inflammation was significantly lower and nitrogen retention significantly higher in animals on the WP diet than in those on the PH diet. Small intestine morphology was maintained with the WP diet, whereas villus height was significantly lower after 66 h, and there were fewer mitoses per crypt in the rats on the PH diet. Glucoamylase activity at all times, and N-aminopeptidase activity at 18 h, were significantly higher in rats on the WP diet. The putrescine (at 42 h) and spermidine (at 18 h) concentrations in the mucosa were higher in the rats on the WP diet. These data suggest that synthetic diets should be tested for their nutritional value during acute inflammation before they are used in human nutrition. PMID- 10378204 TI - Postpartum body composition changes in lactating and non-lactating primiparas. AB - The objective of this study is to evaluate the postpartum body composition changes in lactating versus non-lactating or formula-feeding primiparas during the first 12 wk. Twenty primiparous females (age range 17-35 y) who decided to nurse or formula feed their infant were studied. The non-lactating mothers (n = 6) were younger (21 versus 29 y) and had a lower prepregnancy weight (55 kg versus 63 kg) than the lactating mothers (n = 14). Body weight, height, waist and hip measurements, 3-d dietary and activity records, skin-fold thickness from triceps, suprailiac, midthigh, and midupper arm circumference, and total body composition were evaluated at three time periods (at delivery, at 6 wk, and at 12 wk postpartum). Total body composition for bone mineral, lean, and fat mass was measured by dual energy x-ray absorptiometry. At delivery and 6 wk postpartum, the weights and heights were similar between the two groups. By 12 wk postpartum, the formula-feeding group had a weight loss that was different from delivery, 66 +/- 10 kg to 59 +/- 8 kg, P < 0.03. There was no significant weight change in the lactating group during the study. The weight loss consisted of more lean mass than fat mass. The total body bone mineral content did not differ between the two groups during the study. Both groups had reduction in their waist size from delivery to 12 wk postpartum. But only the non-lactating mothers had reductions in their hip and midthigh measurements. There were no changes between the two groups in the skin-fold measurements. Lactating mothers had a higher total daily calories (1974 +/- 318 versus 1464 +/- 178 calories, P < 0.002) and fat intake (63 +/- 14 versus 47 +/- 9 g, P < 0.02) than the non-lactating mothers. The energy expenditure was similar between both groups. In conclusion, during the first 12 wk postpartum, non-lactating mothers who were younger and weighed less prepregnancy lost body weight and had more waist, hip, and midthigh size reductions compared to lactating mothers. PMID- 10378205 TI - Complication of subcutaneous insertion of Hickman catheter using the Seldinger technique in patient with pendulous breast. PMID- 10378206 TI - Nutritional characteristics of wild primate foods: do the diets of our closest living relatives have lessons for us? AB - The widespread prevalence of diet-related health problems, particularly in highly industrialized nations, suggests that many humans are not eating in a manner compatible with their biology. Anthropoids, including all great apes, take most of their diet from plants, and there is general consensus that humans come from a strongly herbivorous ancestry. Though gut proportions differ, overall gut anatomy and the pattern of digestive kinetics of extant apes and humans are very similar. Analysis of tropical forest leaves and fruits routinely consumed by wild primates shows that many of these foods are good sources of hexoses, cellulose, hemicellulose, pectic substances, vitamin C, minerals, essential fatty acids, and protein. In general, relative to body weight, the average wild monkey or ape appears to take in far higher levels of many essential nutrients each day than the average American and such nutrients (as well as other substances) are being consumed together in their natural chemical matrix. The recommendation that Americans consume more fresh fruits and vegetables in greater variety appears well supported by data on the diets of free-ranging monkeys and apes. Such data also suggest that greater attention to features of the diet and digestive physiology of non-human primates could direct attention to important areas for future research on features of human diet and health. PMID- 10378207 TI - Anorexia and aging: pathophysiology. AB - Aging results in a dysregulation of the ability to regulate food intake. In general, this presents as a decrease in food intake accompanied by early satiation. The early satiation appears to be predominantly due to a decrease in adaptive relaxation of the fundus of the stomach resulting in early antral filling. Increased levels and effectiveness of cholecystokinin also play a role in the anorexia of aging. Leptin levels increase with aging in males but not in females. With regard to the central feeding drive, both the opioid and neuropeptide Y effects appear to decline with age. This physiologic anorexia of aging increases the risk for older persons to develop severe anorexia and weight loss when disease occurs. PMID- 10378208 TI - Liver transplantation, body composition, and substrate utilization: does organ transplantation normalize the metabolic situation of the patient? PMID- 10378209 TI - A non-essential role of liver innervation in controlling feeding behavior. PMID- 10378210 TI - From malnutrition to obesity: changes in nutritional status associated with liver transplantation. PMID- 10378211 TI - Leptin (OB protein) and meal size. PMID- 10378212 TI - Dietary guidelines for the year 2000: epidemiology in action. PMID- 10378213 TI - The effect of medical nutrition therapy on malnutrition and clinical outcomes. PMID- 10378214 TI - Confidence intervals. PMID- 10378215 TI - Missing pieces. PMID- 10378216 TI - Food, nutrition, and the prevention of cancer: a global perspective. American Institute for Cancer Research/World Cancer Research Fund, American Institute for Cancer Research, 1997. PMID- 10378217 TI - Laparoscopy in gynecologic malignancies. AB - One of the cornerstones of gynecologic cancer surgery is the assessment and removal of the retroperitoneal lymph nodes. Numerous reports have demonstrated that, when performed by highly skilled individuals, laparoscopic lymphadenectomies can be performed safely. This has led to the investigation of laparoscopy in the surgical staging and treatment of patients with ovarian, cervical, and endometrial cancers. This very promising approach has the potential to revolutionize numerous aspects of the management of gynecologic malignancies. However, it must be emphasized that the use of laparoscopy for gynecologic malignancies is still in its infancy. Studies that provide complication rates and long-term results are just beginning to be reported. More clinical data are necessary before the laparoscopic techniques are accepted as new surgical standards. Ongoing, prospective clinical trials will help answer many of the questions regarding the safety and efficacy of gynecologic laparoscopy. Until more data accumulate, operative laparoscopy will remain a promising, but unproven, tool in the management of patients with gynecologic malignancies. PMID- 10378218 TI - Immunotherapy in renal cell carcinoma. AB - Patients with metastatic renal cell carcinoma continue to present a therapeutic challenge. Current therapeutic approaches involve surgery and various types of immunotherapy. The rationale for this latter form of therapy include the observations of spontaneous tumor regression, the presence of a T-cell-mediated immune response, and the tumor responses observed in patients receiving cytokine therapy. Analysis of prognostic factors in these patients demonstrates that clinical responses occur most frequently in individuals with good performance status. The cytokines interleukin-2 (IL-2, aldesleukin [Proleukin], interferon alfa (Intron A, Roferon-A), or the combination produce responses in 15% to 20% of patients. Randomized trials suggest that administration of interferon-alfa may result in a modest improvement in median survival. Investigation of the molecular genetics of renal cell carcinoma and the presence of T-lymphocyte immune dysregulation have suggested new therapeutic strategies. Further preclinical and clinical studies investigating inhibitors of angiogenesis or pharmacologic methods to reverse immune dysregulation are ongoing. Therapeutic results in patients with renal cell carcinoma remain limited, and investigational approaches are warranted. PMID- 10378219 TI - Clinical trials. NCI clinical trials in colon cancer. PMID- 10378220 TI - QOL and outcomes research in prostate cancer patients with low socioeconomic status. AB - The VA Cancer of the Prostate Outcomes Study (VA CaPOS) is collecting quality-of life (QOL) information from prostate cancer patients, spouses, and physicians at six VA medical centers. Currently, 601 men with prostate cancer are included in the study, most of whom are of low socioeconomic status and over half of whom are African-American. Quality-of-life responses were most favorable for newly diagnosed patients, intermediate for those with stable metastatic disease, and poorest for those with progressive metastatic disease. Patients could not provide reliable estimates of their own preferences for future QOL states but responded reliably to questions phrased as a comparison of the preferences of two hypothetical patients. High out-of-pocket costs for hormonal therapies, lack of health insurance, and a belief that the non-VA system offered poorer services were the most common reasons for patient transferral to the VA system. Satisfaction with medical care was generally high. While African-American patients were more likely to have advanced prostate cancer at diagnosis, after adjustment for differences in health literacy, race was no longer a significant predictor of advanced disease. The VA CaPOS provides useful information on health status and patient satisfaction of VA prostate cancer patients. Long-term evaluations are needed to detect clinically meaningful QOL information as the disease progresses. PMID- 10378221 TI - Cancer survivors' employment and insurance rights: a primer for oncologists. AB - Cancer survivors' access to equal employment opportunities and adequate health insurance has changed significantly during the 1990s. New federal and state laws have expanded survivors' rights to be treated fairly in the workplace, although some problems still exist. Survivors have experienced less progress, however, in obtaining and retaining adequate health insurance. This article reviews the problems faced by cancer survivors in securing and keeping adequate employment and health insurance. PMID- 10378222 TI - Treatment of estrogen deficiency symptoms in women surviving breast cancer. Part 6: Executive summary and consensus statement. Proceedings of a conference held at Boar's Head Inn, Charlottesville, Virginia, September 21-23, 1997. PMID- 10378223 TI - Modulation of protein kinases and protein phosphatases by reactive oxygen species: implications for hippocampal synaptic plasticity. AB - 1. Reactive oxygen species are known for their role in neurotoxicity. However, recent studies indicate that reactive oxygen species also play a role in cell function under physiological conditions. 2. Both superoxide and hydrogen peroxide alter the activity of various protein kinases and protein phosphatases, some of which are involved in hippocampal synaptic plasticity. Specifically, the activity of protein kinase C, extracellular-regulated kinase 2, and a protein tyrosine kinase(s) is increased in the presence of these reactive oxygen species, whereas the activity of protein phosphatases 2A and 2B, and a protein tyrosine phosphatase(s) is decreased. 3. Protein kinase C, extracellular-regulated kinase 2, and protein tyrosine kinases critically participate in the induction and/or early expression of long-term potentiation at glutamatergic synapses in hippocampus. Protein phosphatases 2A and 2B participate in the induction and/or early expression of long-term depression at these synapses. 4. Treatment of hippocampal slices with scavengers of either superoxide or hydrogen peroxide prevents the full expression of long-term potentiation. Long-term potentiation in hippocampus also is attenuated in transgenic mice that overexpress Cu/Zn superoxide dismutase. 5. The link between reactive oxygen species and long-term potentiation may be the activating effect on protein kinases. The inhibiting effect of reactive oxygen species on protein phosphatases may also contribute to long-term potentiation. 6. The authors hypothesize that reactive oxygen species play a critical role in hippocampal long-term potentiation by favoring the activation of a protein kinase over a protein phosphatase signaling cascade. PMID- 10378224 TI - Understanding the biological activity of amyloid proteins in vitro: from inhibited cellular MTT reduction to altered cellular cholesterol homeostatis. AB - 1. MTT is taken into the cell through endocytosis and is reduced and accumulated in a population of acidic vesicles. Reduced MTT formazan is exocytosed to form needle-like formazan crystals at the cell surface. Mitochondria are unlikely to play a significant role in cellular MTT reduction. 2. Amyloid fibrils inhibit cellular MTT reduction indirectly by enhancing MTT formazan exocytosis. All protein fibrils with beta-pleated sheet structure which have been examined enhance MTT formazan exocytosis and induce neurotoxicity. 3. Cellular free cholesterol regulates the exocytosis of the intracellular MTT formazan transporting vesicles and these vesicles may be involved in cellular cholesterol homeostasis. 4. Amyloid fibrils inhibit cholesterol esterification and alter the distribution of free cholesterol in neurons. 5. Amyloid fibril-induced alterations in cellular cholesterol metabolism and vesicle trafficking may contribute to neurodegeneration in vivo. PMID- 10378225 TI - Representations of knowledge about dominoes in demented and normal elderly players. AB - 1. Dementia patients who retain musical and game-playing skills exhibit impaired performance on explicit memory tests of knowledge about their retained skill. 2. Dementia patients who retain skill at playing dominoes can answer complex questions about the play of the game almost as well as normal elderly domino players when the questions are presented with real dominoes. 3. The aim of this study was to determine if skilled dementia patients could answer questions about domino play when the stimuli were two-dimensional drawings of dominoes. 4. Seventeen dementia patients and eight normal elderly domino players were tested on two forms of the Domino Quiz: first with real dominoes, then with two dimensional drawings; other neuropsychological tests were given at the same time. 5. Fourteen of the 17 patients and all of the controls showed no decline in answering questions about domino play when two-dimensional drawings were used. These patients showed retained symbolic processing of information about dominoes despite declines in overall mental status, generation of words from specific semantic categories, and recognition memory for domino terminology. 6. Because the 14 patients with retained domino skill performed as accurately as controls on both administrations of a letter cancellation task, the ability to process familiar symbols may be important to their game-playing skill. PMID- 10378226 TI - Calbindin immunoreactivity in the hippocampal formation and neocortex of schizophrenics. AB - 1. The authors studied the morphology of CalbindinD28K (CaBp) immunoreactive cells and processes in the hippocampal formation and the prefrontal cortex of schizophrenics using the immunohistochemical technique of avidin-biotin-complex method (ABC method), and the results were compared with those from normal human brains. 2. In the hippocampal formation area CA2 of schizophrenics, many CaBp immunopositive cell bodies and fibers were disordered in their arrangement compared to normal control brains. 3. In the prefrontal cortex (Brodmann area 9) of schizophrenics, many immunopositive cell bodies were exhibited irregular axis arrangement and fiber disarray. 4. The altered distribution pattern of CaBp immunopositive structures in the hippocampal formation and the prefrontal cortex might indicate the existence of GABA(gamma-aminobutyric acid)ergic dysfunction in the brain of schizophrenic patients. PMID- 10378227 TI - P300 and disability of daily life in schizophrenia. AB - 1. The relationship between the P300 component of event-related potentials and disability of daily life was examined in schizophrenia. The subjects were 26 chronic schizophrenic patients. 2. Disability of daily life was assessed by using the Life Assessment Scale for Mental Illness (LASMI). 3. Auditory event-related potentials were recorded during a standard oddball task. 3. P300 amplitude correlated negatively with general psychopathology scale of PANSS (r = -0.416) and Work of LASMI (r = -0.417). P300 latency did not correlate with any of PANSS or LASMI score. 4. These results indicate that P300 amplitude can be an index for disability of daily life in schizophrenia. PMID- 10378228 TI - Screening for depression in the elderly: a study on misclassification by screening instruments and improvement of scale performance. AB - 1. The study compares the psychometric performance of the CES-D and the GHQ-12 in a sample of elderly community residents. Misclassification rates of the questionnaires were analyzed and suggestions for improvement of scale performance are made. 2. 287 subjects out of the general population aged 60-99 years were personally interviewed with standardized diagnostic tools and completed both the GHQ-12 and the CES-D. Best-estimate diagnoses served as standards for receiver operating characteristics (ROC) analysis. 3. Both the GHQ-12 and the CES-D discriminated well between depressive and nondepressive subjects (AUROC = 0.794 and AUROC = 0.782, respectively). The amount of false positive results was high for both questionnaires (GHQ-12: 80.6%, CES-D: 90.1%). Increasing age led to more false positive results on the GHQ-12, whereas the CES-D yielded more false positive results in subjects living in an old age residence or together with family members when compared to those living together with their spouse. 4. The GHQ-12 and the CES-D were valid screening instruments for depression in a community sample of elderly subjects. However, both questionnaires yielded a considerable proportion of false positive results. Elevation of the cut-off score may reduce the misclassification rate of the GHQ-12 but not that of the CES-D. PMID- 10378229 TI - Frontal reactivity and sensation seeking an ERP study in skydivers. AB - 1. In the line of Zuckerman's studies on sensation seeking and optimal level of arousal, the authors hypothesized that high sensation seeking might be used to compensate for anhedonia due to basal arousal deficit. A population of interest was found with parachutists practicing skydiving, generally described as very high sensation seekers. 2. After clinical assessment of emotional and affective components, amplitudes of the frontal P3 of the ERP were used as indices of arousal. 3. Skydivers presented more negative symptoms (anhedonia and blunted affect) than controls. This was observed in isolation from any depressive episode, which would suggest the presence of emotional deficit as a trait. As expected, skydivers presented more sensation seeking than controls. These two results taken together could indicate that sensation seeking is an adaptive reaction to anhedonia. 4. ERP results showed that frontal P3 amplitudes were larger in skydivers than in controls, whereas in a previous study we showed the opposite in depressed patients with a similar emotional deficit. This could indicate that the frontal P3 amplitude does not reflect the emotional deficit per se. We suggest that it rather reflects the capacity to use some behaviors which improve automatic attentional processes in order to obtain arousing stimulation that could counterbalance the emotional deficit. Depressions with emotional deficit might be due to the lack of such a capacity. PMID- 10378230 TI - Alcohol and visual performance. AB - 1. The authors examined the effect of acute alcohol consumption on a set of visual tasks: visual short term memory, depth perception, and attention. 2. In a repeated measurement design, thirteen subjects performed the tasks once sober and once intoxicated with 0.8 g/kg body weight pure ethanol in orange juice (33% alcohol). Subjects underwent a neuropsychological (Benton test) and a psychophysical test (vernier discrimination) both assessing visual short term memory, the test d2 as a measure of attention and concentration, and a psychophysical depth perception task. 3. Subjects demonstrated significant alcohol-related impairments in depth perception and in visual short term memory as assessed by the vernier discrimination task. However, the neuropsychological Benton test and test d2 failed to reveal alcohol-related changes in performance probably due to superimposed learning effects. Performance was neither correlated with blood alcohol levels (BAL) nor perceived intoxication. Even though the current BAL was known to the subjects, only half of them demonstrated a close correlation between BAL and perceived intoxication. PMID- 10378231 TI - Effect of acute and chronic tramadol on [3H]-norepinephrine-uptake in rat cortical synaptosomes. AB - 1 Tramadol hydrochloride is a centrally acting opioid analgesic whose efficacy and potency is only five to ten times lower than that of morphine. Opioid, as well as non-opioid mechanisms, may participate in the analgesic activity of tramadol. 2 [3H]-NE uptake in isolated rat cortical synaptosomes was studied in the presence of tramadol, desipramine, methadone, and morphine. Desipramine and tramadol inhibited synaptosomal [3H]-NE uptake with apparent Kis of 7.3 +/- 0.66 and 1.4 +/- 0.0045 microM, respectively. Methadone was active at a 10-fold higher concentration (Ki: 87 +/- 5.6 microM). In contrast, morphine essentially failed to inhibit [3H]-5-HT uptake (Ki: 0.75 +/- 0.40 M). 3 Methadone, morphine, and tramadol were active in the hot plate test with ED50s of 6.2, 9.3, and 40 mg kg 1, respectively. 4 [3H]-NE uptake was examined in synaptosomes prepared from rats 30 min after receiving a single dose of morphine, methadone or tramadol. Only tramadol (31 mg kg-1, i.p.) decreased uptake of the transmitter, with an ED50 equal to that in the hot plate test. 5 Animals were chronically treated for 15 days with increasing doses of tramadol (20 to 125 mg kg-1, i.p.). Twenty-four hours after the last drug injection, a challenge dose of tramadol (40 mg kg-1, i.p.) was administered. Chronic tramadol was still able to reduce [3H]-NE uptake by 35%. 6 These results further support the hypothesis that [3H]-NE uptake inhibition may contribute to the antinociceptive effects of tramadol. The lack of tolerance in [3H]-NE uptake, together with the absence of behavioural alteration after chronic tramadol treatment proposes that tramadol holds potential over classical opioids in the treatment of pain disorders. PMID- 10378232 TI - Involvement of 5-HT3 receptors in the prolactin release induced by immobilization stress in rats. AB - 1. Central serotoninergic participation in prolactin (PRL) secretion regulation is a well-established event. Furthermore, 5-HT may participate in the mediation of stress-induced PRL release. 2. The authors investigated the effect of the blockade of 5-HT3 receptors on the PRL release induced by immobilization stress in male rats. 3. Pretreatment with the 5-HT3 receptor antagonist, ondansetron (0.10-1.0 mg/kg i.p.), inhibited about 50% of the PRL response to immobilization stress. A lower dose of ondansetron (0.05 mg/kg i.p.) had no effect on the PRL response. No dose-response inhibition was observed. 4. It is concluded that serotoninergic activation related to the mediation of the stress-induced PRL response involves the 5-HT3 receptors. PMID- 10378233 TI - Anticipatory responding, exclusive drug-context pairing and conditioned effects in sensitization to apomorphine-induced climbing in mice. AB - 1. The conditioning aspects of contextual sensitization were examined in the case of apomorphine-induced wall-climbing in mice, measuring onset latencies of the pharmacological response and controlling differential habituation to the test context during drug treatment. 2. Sensitization was generated in male out-bred mice which received intermittent i.p. injections of 0.4 mg/kg apomorphine over 9 daily sessions. On day 10, they were tested for contextual sensitization (all mice under apomorphine). On day 14, after 3 sessions of reinstatement, mice were tested for conditioned climbing (all mice under saline). 3. It was found that simultaneous exposure to both apomorphine and the test context facilitated the expression of a full-blown contextual sensitization (some non-contextual sensitization emerging too); importantly, sensitization was accompanied by a progressive shortening of the latencies to climb (before injections); conditioned climbing appeared only in mice pairing the drug with the test context, that response being absent in mice treated outside the context or never exposed to the context. 4. It is likely that contextual sensitization to apomorphine-induced climbing relies on Pavlovian conditioning processes rather than on habituation related processes. PMID- 10378234 TI - Synaptotagmin and synaptic transmission alterations in apolipoprotein E-deficient mice. AB - 1. Aged apoE-deficient mice and age-matched controls were tested for cognitive alterations in the Morris water maze. 2. Water maze results were correlated with in vivo electrophysiology and expression of the synaptic protein synaptotagmin (p65). 3. Compared to age-matched controls, apolipoprotein E-deficient mice displayed significant performance impairment accompanied by in vivo electrophysiological alterations in the dentate gyrus. 4. Apolipoprotein E deficient mice also showed a significant increase in the synaptic protein, synaptotagmin, a synaptic calcium sensor involved in neurotransmitter release. 5. Cognitive impairments in these animals may be associated with decreased synaptic excitability in hippocampal neurons and the regulatory role of apolipoprotein E in synaptic function might be mediated by modulation of the expression of calcium sensor proteins. PMID- 10378235 TI - Selectivity of action of typical and atypical anti-psychotic drugs as antagonists of the behavioral effects of 1-[2,5-dimethoxy-4-iodophenyl]-2-aminopropane (DOI). AB - 1. There has been considerable research in the field of schizophrenia over the past few years with emphasis on the discovery of better drugs, particularly those with 5-HT2 antagonist activity. 2. In an effort to enhance identification of such compounds and to further understand the contribution of 5-HT2 activity to the effects of antipsychotic drugs, a series of conventional, atypical and purported antipsychotic compounds were assessed as antagonists of DOI-induced behaviors in rats. 3. DOI (1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane hydrochloride) is an hallucinogen having high affinity and selectivity as an agonist at 5-HT2A/2C receptors. Over a 30-min period after injection, DOI (0.3-10.0 mg/kg; i.p.) produced dose-related behavioral effects including head-and-body shakes, forepaw tapping and skin-jerks. Effects of the antipsychotic drugs and other compounds (30 min pretreatment; i.p.) were examined against a fixed dose of DOI (3.0 mg/kg). 4. In a dose-dependent manner, M100907 (MDL 100,907), risperidone, haloperidol, clozapine, iloperidone, olanzapine, amperozide, remoxipride, ritanserin and the neurotensin agonist NT1 (N alpha MeArg-Lys-Pro-Trp-Tle-Leu) antagonized each of the three behavioral effects of DOI. Drugs attenuating the head-and-body shakes were equally effective in blocking both forepaw tapping and skin-jerks indicating that these behaviors are mediated by similar mechanisms. The following compounds had either inconsistent or no effect on the DOI-induced behaviors: SB 200646A, citalopram, imipramine, fluoxetine, morphine, CP 99994, diazepam, ondansetron and SKF 97541. 5. The data show that antipsychotic agents, as a drug class, effectively block the effects of DOI. These actions are selective, as a series of nine non-antipsychotic and centrally-acting drugs were generally inactive in the procedure. PMID- 10378236 TI - Combined electroconvulsive therapy and clozapine in treatment-resistant schizophrenia. AB - 1. To assess the efficacy and safety of combining electroconvulsive therapy (ECT) and clozapine in patients with treatment-resistant schizophrenia, the authors reviewed use of this combination in four treatment-resistant schizophrenic inpatients and one inpatient with schizophrenia who was intolerant of clozapine doses needed to control her psychosis. 2. The combination of clozapine and bilateral ECT was modestly effective in two treatment-resistant patients and markedly effective in three patients. There was significant overall improvement in patients' Clinical Global Impression (CGI) and Global Assessment of Functioning (GAF) scores (p < 0.005 and p < 0.0004, respectively), however in patients where marked symptomatic improvement was noted, effects were not sustained. 3. One of the patients that showed dramatic yet transient improvement followed by relapses received maintenance ECT but relapsed despite this. 4. The authors saw no adverse effects in connection with the combination of ECT and clozapine. 5. Supplementing clozapine with a course of bilateral ECT appears to be safe and is effective in some patients with refractory schizophrenia, however its beneficial effects may be short-lived. The long-term impact of ECT on the course of schizophrenia in patients incompletely responsive to clozapine is not fully elucidated. PMID- 10378237 TI - Do panic disorder and posttraumatic stress disorder share a common psychoneuroendocrinology? PMID- 10378238 TI - Association of depressiveness with blunted growth hormone response to maximal physical exercise in young healthy men. AB - Blunted response of growth hormone secretion to several pharmacological challenges is present in depression, but much less is known about the relationship of depression and secretion of growth hormone elicited by physiological stimuli. Furthermore, it is not known whether blunted growth hormone response occurs in depressiveness as measured with psychometric scales. A total of 82 healthy male volunteers (age 18-26 years) exercised on a bicycle ergometer with incremental load to achieve their maximal performance. Before exercise, subjects filled in approbated versions of Beck Depression Inventory (BDI), Spielberger's State-Trait Anxiety Scale, Cohens Perceived Stress Scale, and Schwartzers Self-Efficacy Scale. Blood samples were collected before and after exercise, and growth hormone, cortisol, and testosterone were measured by chemiluminescence immunoassay. Median perceived stress score of the subjects was identical to our population-based database median value, but the subjects had higher self-efficacy and lower depressiveness as shown by median values. In the majority of subjects, physical exercise induced remarkable increases in blood levels of the hormones. Cortisol and testosterone levels were not associated with the scores of psychometric scales. However, growth hormone response was virtually absent in high scorers (above median population score, n = 24) in BDI total score and the negative attitude subcomponent. Hence, this study demonstrates that growth hormone response to physiological stimuli is reduced in psychometrically measured depressiveness. PMID- 10378239 TI - Cortisol levels of young children in full-day childcare centers: relations with age and temperament. AB - Cortisol levels of 70 children, aged 39-106 months, were sampled at home and at their full-day childcare centers at two times of day, mid-morning and mid afternoon. Parents and teachers completed questionnaires assessing child temperament (negative affectivity, surgency or extroversion, and effortful control) and aggressive behavior. The results replicated a previous study showing increases in cortisol levels over the day at childcare for preschool-aged children, while home levels followed the expected circadian decrease in cortisol from morning to afternoon for most children regardless of age. At childcare, 3- and 4-year olds were more likely to show elevations in cortisol by mid-afternoon than were older children. Controlling statistically for age, shyness for boys, and poor self-control and aggression for both sexes were associated with increases in cortisol over the day at childcare. The results suggest that younger children and those with more immature social skills may frequently experience elevations in cortisol as the day progresses in group care contexts. PMID- 10378240 TI - Cognitive-behavioral stress management buffers decreases in dehydroepiandrosterone sulfate (DHEA-S) and increases in the cortisol/DHEA-S ratio and reduces mood disturbance and perceived stress among HIV-seropositive men. AB - This study examined the effects of a 10-week cognitive-behavioral stress management (CBSM) intervention on dehydroepiandrosterone sulfate (DHEA-S) levels and the ratio of cortisol to DHEA-S (cortisol/DHEA-S), potential surrogate adrenal markers of HIV disease progression, in relation to alterations in mood and distress. HIV-seropositive men were randomized to either a group-based CBSM intervention (n = 43) or to a wait-list control group (n = 24), with both hormonal and distress measures assessed just prior to and immediately following the 10-week period. Results showed that CBSM buffers decreases in DHEA-S and increases in the cortisol/DHEA-S ratio. Further examination also revealed that changes in the cortisol/DHEA-S ratio were significantly and positively related to changes in total mood disturbance and perceived stress over time. These findings demonstrate that a short-term CBSM intervention can buffer against decrements in DHEA-S and increments in the cortisol/DHEA-S ratio among symptomatic, HIV positive men, and that alterations in the cortisol/DHEA-S ratio move in concert with changes in mood and distress observed during CBSM. PMID- 10378241 TI - Effects of competition and its outcome on serum testosterone, cortisol and prolactin. AB - In various species, competitive encounters influence hormonal responses in a different way depending on their outcome, victory or defeat. This study aimed to investigate the effects of sports competition and its outcome on hormonal response, comparing it with those displayed in situations involving non-effort and non-competitive effort. To this end, serum testosterone (T), cortisol (C) and prolactin (PRL) were measured in 26 judoists who participated in three sessions (control, judo fight and ergometry). The relationship between hormonal changes and psychological variables before and after the fight were also analysed. Our results showed a hormonal response to competition, which was especially characterized by an anticipatory rise of T and C. Depending on outcome, significant higher C levels were found in winners in comparison to losers through all the competition but not in T or PRL, both groups expending a similar physical effort. Furthermore, similar hormonal responses to the fight and to a non competitive effort with the same caloric cost were found, other than with PRL. Winners showed a higher appraisal of their performance and satisfaction with the outcome, and perceived themselves as having more ability to win than losers, although there were no significant differences in motivation to win. Finally, the relationships found between T changes in competition and motivation to win, as well as between C response and self-efficacy suggest that in humans hormonal response to competition is not a direct consequence of winning and losing but rather is mediated by complex psychological processes. PMID- 10378242 TI - Assessing dehydroepiandrosterone in saliva: a simple radioimmunoassay for use in studies of children, adolescents and adults. AB - While salivary assays for some hormones are widely used, the availability of assays for salivary DHEA is limited. By adapting a commercially available radioimmunoassay serum kit, we developed a reliable, efficient and sensitive measure of DHEA in saliva that does not require separation or extraction. The minimum detection limit was 4.0 pg/ml. Intra-assay coefficients of variation (CV%) were on average 4.05, and inter-assay CVs averaged 9.70. Method accuracy, determined by spike recovery, and linearity, determined by serial dilution, averaged 99.55 and 92.03%. Levels in matched serum and saliva samples showed strong linear relationships for adult males and females. Specific guidelines are developed for sample collection, storage, and preparation procedures. Reference ranges for salivary DHEA levels are provided for 64 children ages 8-11, 96 adolescents ages 12-17 and 48 adults ages 30-45. Salivary DHEA levels are shown to reflect developmental, gender and diurnal differences. PMID- 10378243 TI - [Medical guidelines: the legal implications]. AB - The author discusses the legal implications of medical guidelines. If treatment differs from such guidelines and the results for the patient are negative, the physician will come under considerable pressure to explain why the treatment was correct lege artis, deviation from guidelines notwithstanding. Sense or nonsense of guidelines is a matter to be discussed among the medical profession. However, if guidelines should attempt to limit or lower the quality of care for financial reasons, we are faced with the issue of priorities in a world where resources are limited. This is a legal issue which society as a whole has to decide; it is not a matter to be left to medical experts. PMID- 10378244 TI - [Consensus conferences and guidelines in gastroenterology: an illusion]. AB - Evidence-based medicine describes the present-day situation of a rational medicine which has its origins in the 19th century. At present evidence-based medicine is an ideal which is unattainable due to inadequate data from randomised controlled studies. Guidelines and consensus conferences which are not based on clearcut scientific data from controlled prospective and randomised studies fail to meet the requirements of evidence-based medicine and so cannot improve patient management. Guidelines and consensus conferences do not substantially reduce health costs. PMID- 10378245 TI - [Consensus conferences and guidelines in gastroenterology: a valid approach]. AB - The rapid and continuous increase in knowledge has meant that medicine has become more complex and involved, and at the same time less transparent, for physicians and patients. To ensure quality and improve patients confidence, new structures in quality management are necessary. Guidelines, standards and consensus conferences by experts are valid ways of ensuring the future quality of medical care. PMID- 10378246 TI - [Diagnosis and treatment of common bile duct stones: a current review and the Berne concept]. AB - The prevalence of common bile duct stones increases with age. Whereas patients under 60 show a prevalence of 8-15%, the rate increases up to 15-60% > 60 years. The main goal of optimal preoperative diagnosis and treatment of bile duct stones is to avoid unnecessary and negative investigations and to decrease the rate of retained/missed stones. Anamnestic and clinical pointers, laboratory findings and "simple" radiological examinations (ultrasonography, intravenous cholangiography) allow selective preoperative ERCP. The reported clearance rates for ERCP are as high as 95%, with a morbidity and mortality of 3-15% and 0.2-2.5% respectively. Only after endoscopic stone removal should laparoscopic cholecystectomy be added. Laparoscopic bile duct exploration is technically demanding and offers no objective advantages compared to ERCP. PMID- 10378247 TI - [Variceal hemorrhage in portal hypertension: role of surgery in the acute and elective situation]. AB - The role of surgery in portal hypertension has changed over time. The past decade has seen significant advances in pharmacotherapy (acute and elective), endoscopy and interventional radiology. However, mortality from the first bleeding remains constant between 30 and 50% and depends directly on patient risk (Child C). Surgical intervention during the acute bleeding phase carries a mortality rate of up to 70% and should therefore be avoided. About 90% of patients with acute variceal haemorrhage may satisfactorily be managed with pharmacotherapy and/or endoscopic banding alone. If bleeding persists, balloon tamponade (Linton) is indicated. In case of recurrent bleeding under maximal therapy (problem bleeder), delayed shunting may be indicated. In patients with Child A/B cirrhosis surgical mesocaval shunt with an interposition graft is preferred, whereas for transplant candidates a TIPS is used. The long-term outcome for surgical shunts is significantly better compared to TIPS. Secondary prophylaxis consists of medical treatment (propanolol) and repeated endoscopic banding. If rebleeding occurs under adequate therapy, surgery (mesocaval shunt/TIPS) should be evaluated. However, liver transplantation is the only curative therapeutic option for this life-threatening disease. PMID- 10378248 TI - [Gold induced cystitis]. PMID- 10378249 TI - Squamous cell carcinoma of the sinonasal cavities. AB - Squamous cell carcinoma of the sinonasal cavity is an uncommon neoplasm. The symptoms of this tumor are aspecific, and the diagnosis is often made in an advanced stage of disease. Imaging is necessary for evaluating paranasal extension and possible intracranial spread, orbital involvement, infratemporal extension, spread to the nasopharynx, oropharynx, or oral cavity, and spread along neurovascular bundles or perineural spread. Sinonasal cancers are studied with CT or MRI, and in many cases both modalities are used, as they often offer complementary information. The CT and MRI appearance of this kind of tumor is reviewed in this article. PMID- 10378250 TI - Radiologic evaluation of neck metastases: the otolaryngologist's perspective. AB - Imaging of the neck for the assessment of nodal metastases can be used to detect occult metastases or to assess operability of palpable metastases. The detection of small occult metastases has limitations, as micrometastases cannot be depicted. However, imaging can diminish the risk of occult metastases and thus influence management; for this purpose a very sensitive technique is necessary. The currently used radiologic criteria are not sensitive enough to accomplish enough reduction in the risk of occult metastases. Therefore, more sensitive CT and MRI criteria, but especially ultrasound guided aspiration, should be used to assess the clinically negative neck. PMID- 10378251 TI - Imaging of perineural tumor spread in head and neck cancer. AB - Perineural spread (PNS) of head and neck tumors is a well-described phenomenon whereby a lesion can migrate away from the primary site along the neural sheath. By this mechanism, tumor can spread a considerable distance and compromise vital neurologic structures, with significant impact on treatment and prognosis. It is crucial that radiologists search for PNS whenever imaging the head and neck cancer patient. Detection of PNS requires familiarity with common tumor types and locations that can give rise to it, an understanding of the relevant cranial nerve anatomy, and an awareness of the radiologic appearance and techniques best suited to imaging this important and often overlooked complication of head and neck cancer. PMID- 10378252 TI - Application of new imaging techniques for the evaluation of squamous cell carcinoma of the head and neck. AB - The past several years have seen dramatic changes in imaging of the head and neck. Technical improvements in CT and MRI coupled with their widespread availability have made cross-sectional imaging an important adjunct in evaluation of patients with disease of the extracranial head and neck. The most recent advances in head and neck imaging are from new metabolic and functional imaging techniques. The intent of this report is to provide an update on the potential role of positron emission tomography, new MRI agents, and magnetic resonance spectroscopy for evaluating squamous cell carcinoma of the extracranial head and neck. PMID- 10378253 TI - Multivariate competing risks. AB - Competing risks models can be used to compare the effect of risk factors for different causes of death or subtypes of a disease. However, sometimes more than one outcome classification is available and if two such classifications are correlated, one may speculate whether differences in the effect of a risk factor according to one classification simply may be an effect of differences according to the other correlated classification. We introduce in this paper the new concept of multivariate competing risks to test formally such a hypothesis. PMID- 10378254 TI - Is the AIDS incubation time changing? A back-calculation approach. AB - Contradictory literature was recently published on possible changes in AIDS incubation time over the period 1978-1994. The purpose of this work was to test if a change in incubation time (shortening or lengthening) was observed in France, either globally or in specific transmission groups (homosexual-bisexual men, heterosexual subjects), using a back-calculation approach. An age dependent TSI model (time since infection), which took into account a temporary treatment effect and allowed us to test for a change in the incubation time, was applied to the French AIDS cases (Reseau National de Sante Publique). The EM algorithm was used to maximize the likelihood and the best model was selected on the basis of the likelihood ratio statistic. The analysis on all AIDS cases indicated a shortening of the AIDS incubation time estimated to begin in 1983 (95 per cent CI 1982-1984). This shortening of incubation time was also apparent when analysis was restricted to homosexual-bisexual men and to heterosexual subjects. This shortening corresponded to a median incubation time of 9.6 years (95 per cent CI 8.1-10.5) for people infected at 30 years of age in 1983, versus 12.7 years for people infected at 30 years of age before the change. PMID- 10378255 TI - Group testing in presence of classification errors. AB - We modify Dorfman's and Sterrett's group testing protocols to make them suitable for testing blood samples for the presence of HIV antibodies, as well as for many industrial applications, when false negatives cannot be tolerated. We first propose that test kit sensitivity be increased to nearly 100 per cent by altering the reactive versus non-reactive threshold. Subsequently, group and repeat testing are used with a careful selection of group size and the number of times a test is repeated, in order to maximize efficiency while keeping the false positive predictive value (FPPV) within a specified limit. Numerical calculations show that our testing protocol is efficient, has low procedural complexity and keeps both types of classification errors below specified tolerance limits. PMID- 10378256 TI - Adjusting for measurement error to assess health effects of variability in biomarkers. Multicenter AIDS Cohort Study. AB - Longitudinal studies of health effects often relate individuals' biomarker levels to disease progression. Repeated measurements also provide an opportunity to assess within-individual biomarker variability, and it is reasonable to postulate that this measure might provide additional information about a particular outcome variable. Given the existing precedent for application of adjustment methods to account for measurement error in subject-specific average levels of a covariate, this concept motivates the application of such methods to incorporate variability as well. In this paper, we investigate the nature of the relationship between the decline of CD4 cell count induced by infection with human immunodeficiency virus, and CD4 level and variability prior to infection. We first describe the distribution of repeated CD4 measurements prior to infection using a model that accounts both for random average levels and random subject-specific variance components. Based on this model, we define true unobservable random variables that correspond to prior level and stability. We perform a linear regression analysis, using these latent variables as covariates, by means of a full maximum likelihood approach. We compare the resulting parameter estimates with those based on regressions employing sample-based estimates of pre-infection levels and variances, and empirical Bayes estimates of these quantities. Although the final inferences are similar to those based on the unadjusted analysis, we find that the magnitude of association with prior level decreases, while that with prior stability increases. Stratified analyses indicate that smoking status affects the relationship between prior CD4 level and initial CD4 decline. We point out advantages associated with the maximum likelihood approach in this particular application. PMID- 10378258 TI - Dichotomization of continuous measurements using generalized additive modelling- application in predicting intrapartum caesarean delivery. AB - In prediction model development, continuous variables are often dichotomized at empirically chosen thresholds to simplify calculations and facilitate decision making. However, these choices are often made in the absence of estimated covariate effects and may be inaccurate, thus weakening the models. To improve this approach, generalized additive modelling that allows non-parametric estimation of the true covariate effects is used for threshold selection. In this study, this approach is illustrated by development of prediction models for intrapartum Caesarean deliveries. The prediction performance of the models thus developed is significantly better than that developed using empirically chosen thresholds for dichotomization. PMID- 10378257 TI - Interval estimation of the common odds ratio from k(2 x 2) tables under cluster sampling. AB - We propose a simple correction factor for the variance of the logarithm of the common odds ratio estimated by the method of Mantel and Haenszel from a series of (2 x 2) tables when data are cluster correlated. The adjustment is applied to the variance estimators proposed by Hauck and by Robins, Breslow and Greenland for the log of the Mantel-Haenszel common odds ratio, and its performance is evaluated in a simulation study. The key features of the proposed adjustment are: (i) it has closed-form; (ii) it can accommodate covariates defined at the cluster specific level, the site-specific level, or both; and (iii) it does not require the user to specify a particular correlation structure for the response data. The correction derives from Liang and Zeger's generalized estimating equations (GEE) technique for logistic regression modelling. Via simulation, we examine empirical versus nominal coverage probabilities for interval estimation of the common odds ratio using adjusted and unadjusted variance estimates, and we present ratios of observed to estimated variances. Results are compared to those obtained from the fully iterated GEE analysis. The characteristics of the simulation study mimic scenarios common in the periodontal research setting, with small numbers of subjects (N = 25, 50), moderate numbers of sites per cluster (m = 4, 16, 32), and modest intracluster correlation levels (rho = 0.0, 0.1, 0.2, 0.3). Results show that adjusted confidence intervals (applied to the Hauck or the Robins, Breslow, Greenland variance estimate) provide coverage probabilities close to the nominal level for the Mantel--Haenszel common odds ratio over a variety of cluster sizes and levels of correlation. PMID- 10378259 TI - A transition rate model for first admissions to psychiatric institutions. AB - This paper describes the application of a parametric transition rate model, the generalized log-logistic model, to the duration of first admissions to psychiatric institutions. The final model included diagnosis, gender, age, living conditions and year of admission as covariates. Characteristics of the log logistic model are described extensively. Parametric transition models offer challenging and promising possibilities to model duration of hospital stay. PMID- 10378260 TI - Mixtures of proportional hazards regression models. AB - This paper presents a mixture model which combines features of the usual Cox proportional hazards model with those of a class of models, known as mixtures-of experts. The resulting model is more flexible than the usual Cox model in the sense that the log hazard ratio is allowed to vary non-linearly as a function of the covariates. Thus it provides a flexible approach to both modelling survival data and model checking. The method is illustrated with simulated data, as well as with multiple myeloma data. PMID- 10378261 TI - Comparing treatment variances in repeated measures bioavailability trials. AB - In repeated measures bioavailability/bioequivalence studies involving two formulations, the underlying measurements, such as the area under the plasma concentration-time curve (AUC) and the maximum plasma concentration (Cmax), typically have a block compound-symmetric covariance matrix. To test the equality of the variances in such a covariance matrix, we derive in this paper an exact test and four asymptotic tests. We compare these tests using simulation to determine those that have good performance in terms of controlling the type I error rate as well as providing good power. We provide an example to demonstrate the proposed method. PMID- 10378262 TI - A method for determining the size of internal pilot studies. PMID- 10378263 TI - Turn up the HEAT. AB - The recently determined crystal structure of the PR65/A subunit of protein phosphatase 2A reveals the architecture of proteins containing HEAT repeats. The structural properties of this solenoid protein explain many functional characteristics and account for the involvement of solenoids as scaffold, anchoring and adaptor proteins. PMID- 10378264 TI - Channel gate! Tension, leak and disclosure. AB - The crystal structure of a bacterial MscL shows how this homopentameric channel protein is held tightly shut to prevent leakage whilst at rest. By inference, the structure also shows how a stretch force in the lipid bilayer causes the channel to open. We now have a concrete picture as to how a stimulus 'gates' an ion channel. PMID- 10378265 TI - Computational protein design. AB - A 'protein design cycle', involving cycling between theory and experiment, has led to recent advances in rational protein design. A reductionist approach, in which protein positions are classified by their local environments, has aided development of an appropriate energy expression. The computational principles and practicalities of the protein design cycle are discussed. PMID- 10378266 TI - Demystifying the synchrotron trip: a first time user's guide. PMID- 10378267 TI - Structural details of urea binding to barnase: a molecular dynamics analysis. AB - BACKGROUND: The molecular mechanism of urea-induced protein unfolding has not been established. It is generally thought that denaturation results from the stabilizing interactions of urea with portions of the protein that are buried in the native state and become exposed upon unfolding of the protein. RESULTS: We have performed molecular dynamics simulations of barnase (a 110 amino acid RNase from Bacillus amyloliquefaciens) with explicit water and urea molecules at 300 K and 360 K. The native conformation was unaffected in the 300 K simulations at neutral and low pH. Two of the three runs at 360 K and low pH showed some denaturation, with partial unfolding of the hydrophobic core 2. The first solvation shell has a much higher density of urea molecules (water/urea ratio ranging from 2.07 to 2.73) than the bulk (water/urea ratio of 4.56). About one half of the first-shell urea molecules are involved in hydrogen bonds with polar or charged groups on the barnase surface, and between 15% and 18% of the first shell urea molecules participate in multiple hydrogen bonds with barnase. The more stably bound urea molecules tend to be in crevices or pockets on the barnase surface. CONCLUSIONS: The simulation results indicate that an aqueous urea solution solvates the surface of a polypeptide chain more favorably than pure water. Urea molecules interact more favorably with nonpolar groups of the protein than water does, and the presence of urea improves the interactions of water molecules with the hydrophilic groups of the protein. The results suggest that urea denaturation involves effects on both nonpolar and polar groups of proteins. PMID- 10378268 TI - Crystal structure of 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase, a potential target for the development of novel antimicrobial agents. AB - BACKGROUND: Folate cofactors are essential for life. Mammals derive folates from their diet, whereas most microorganisms must synthesize folates de novo. Enzymes of the folate pathway therefore provide ideal targets for the development of antimicrobial agents. 6-Hydroxymethyl-7,8-dihydropterin pyrophosphokinase (HPPK) catalyzes the transfer of pyrophosphate from ATP to 6-hydroxymethyl-7,8 dihydropterin (HP), the first reaction in the folate biosynthetic pathway. RESULTS: The crystal structure of HPPK from Escherichia coli has been determined at 1.5 A resolution with a crystallographic R factor of 0.182. The HPPK molecule has a novel three-layered alpha beta alpha fold that creates a valley approximately 26 A long, 10 A wide and 10 A deep. The active center of HPPK is located in the valley and the substrate-binding sites have been identified with the aid of NMR spectroscopy. The HP-binding site is located at one end of the valley, near Asn55, and is sandwiched between two aromatic sidechains. The ATP binding site is located at the other end of the valley. The adenine base of ATP is positioned near Leu111 and the ribose and the triphosphate extend across and reach the vicinity of HP. CONCLUSIONS: The HPPK structure provides a framework to elucidate structure/function relationships of the enzyme and to analyze mechanisms of pyrophosphoryl transfer. Furthermore, this work may prove useful in the structure-based design of new antimicrobial agents. PMID- 10378269 TI - Crystal structure of an aminoglycoside 6'-N-acetyltransferase: defining the GCN5 related N-acetyltransferase superfamily fold. AB - BACKGROUND: The predominant mechanism of antibiotic resistance employed by pathogenic bacteria against the clinically used aminoglycosides is chemical modification of the drug. The detoxification reactions are catalyzed by enzymes that promote either the phosphorylation, adenylation or acetylation of aminoglycosides. Structural studies of these aminoglycoside-modifying enzymes may assist in the development of therapeutic agents that could circumvent antibiotic resistance. In addition, such studies may shed light on the development of antibiotic resistance and the evolution of different enzyme classes. RESULTS: The crystal structure of the aminoglycoside-modifying enzyme aminoglycoside 6'-N acetyltransferase type li (AAC(6')-li) in complex with the cofactor acetyl coenzyme A has been determined at 2.7 A resolution. The structure establishes that this acetyltransferase belongs to the GCN5-related N-acetyltransferase superfamily, which includes such enzymes as the histone acetyltransferases GCN5 and Hat1. CONCLUSIONS: Comparison of the AAC(6')-li structure with the crystal structures of two other members of this superfamily, Serratia marcescens aminoglycoside 3-N-acetyltransferase and yeast histone acetyltransferase Hat1, reveals that of the 84 residues that are structurally similar, only three are conserved and none can be implicated as catalytic residues. Despite the negligible sequence identity, functional studies show that AAC(6')-li possesses protein acetylation activity. Thus, AAC(6')-li is both a structural and functional homolog of the GCN5-related histone acetyltransferases. PMID- 10378270 TI - Crystal structure of 7,8-dihydroneopterin triphosphate epimerase. AB - BACKGROUND: Dihydroneopterin triphosphate (H2NTP) is the central substrate in the biosynthesis of folate and tetrahydrobiopterin. Folate serves as a cofactor in amino acid and purine biosynthesis and tetrahydrobiopterin is used as a cofactor in amino acid hydroxylation and nitric oxide synthesis. In bacteria, H2NTP enters the folate biosynthetic pathway after nonenzymatic dephosphorylation; in vertebrates, H2NTP is used to synthesize tetrahydrobiopterin. The dihydroneopterin triphosphate epimerase of Escherichia coli catalyzes the inversion of carbon 2' of H2NTP. RESULTS: The crystal structure of the homo octameric protein has been solved by a combination of multiple isomorphous replacement, Patterson search techniques and cyclic averaging and has been refined to a crystallographic R factor of 18.8% at 2.9 A resolution. The enzyme is a torus-shaped, D4 symmetric homo-octamer with approximate dimensions of 65 x 65 A. Four epimerase monomers form an unusual 16-stranded antiparallel beta barrel by tight association between the N- and C-terminal beta strands of two adjacent subunits. Two tetramers associate in a head-to-head fashion to form the active enzyme complex. CONCLUSIONS: The folding topology, quaternary structure and amino acid sequence of epimerase is similar to that of the dihydroneopterin aldolase involved in the biosynthesis of the vitamin folic acid. The monomer fold of epimerase is also topologically similar to that of GTP cyclohydrolase I (GTP CH-1), 6-pyrovoyl tetrahydropterin synthase (PTPS) and uroate oxidase (UO). Despite a lack of significant sequence homology these proteins share a common subunit fold and oligomerize to form central beta barrel structures employing different cyclic symmetry elements, D4, D5, D3 and D2, respectively. Moreover, these enzymes have a topologically equivalent acceptor site for the 2-amino-4-oxo pyrimidine (2-oxo-4-oxo pyrimidine in uroate oxidase) moiety of their respective substrates. PMID- 10378271 TI - The 2.7 A crystal structure of deoxygenated hemoglobin from the sea lamprey (Petromyzon marinus): structural basis for a lowered oxygen affinity and Bohr effect. AB - BACKGROUND: The hemoglobins of the sea lamprey are unusual in that cooperativity and sensitivity to pH arise from an equilibrium between a high-affinity monomer and a low-affinity oligomer. Although the crystal structure of the monomeric cyanide derivative has previously been determined, the manner by which oligomerization acts to lower the oxygen affinity and confer a strong Bohr effect has, until now, been speculative. RESULTS: We have determined the crystal structure of deoxygenated lamprey hemoglobin V by molecular replacement to 2.7 A resolution, in a crystal form with twelve protomers in the asymmetric unit. The subunits are arranged as six essentially identical dimers, with a novel subunit interface formed by the E helices and the AB corner using the standard hemoglobin helical designations. In addition to nonpolar interactions, the interface includes a striking cluster of four glutamate residues. The proximity of the interface to ligand-binding sites implicates a direct effect on ligand affinity. CONCLUSIONS: Comparison of the deoxy structure with that of the cyanide derivative revealed conformational changes that appear to be linked to the functional behavior. Oligomerization is coupled with a movement of the first half of the E helix by up to 1.0 A towards the heme, resulting in steric interference of ligand binding to the deoxy structure. The Bohr effect seems to result from proton uptake by glutamate residues as they are buried in the interface. Unlike human and mollusc hemoglobins, in which modulation of function is due to primarily proximal effects, regulation of oxygen affinity in lamprey hemoglobin V seems to depend on changes at the distal (ligand-binding) side of the heme group. PMID- 10378272 TI - The allosteric transition of glucosamine-6-phosphate deaminase: the structure of the T state at 2.3 A resolution. AB - BACKGROUND: The allosteric hexameric enzyme glucosamine-6-phosphate deaminase from Escherichia coli catalyses the regulatory step of N-acetylglucosamine catabolism, which consists of the isomerisation and deamination of glucosamine 6 phosphate (GlcN6P) to form fructose 6-phosphate (Fru6P) and ammonia. The reversibility of the catalysis and its rapid-equilibrium random kinetic mechanism, among other properties, make this enzyme a good model for studying allosteric processes. RESULTS: Here we present the structure of P6(3)22 crystals, obtained in sodium acetate, of GlcN6P deaminase in its ligand-free T state. These crystals are very sensitive to X-ray radiation and have a high (78%) solvent content. The activesite lid (residues 162-185) is highly disordered in the T conformer; this may contribute significantly to the free-energy change of the whole allosteric transition. Comparison of the structure with the crystallographic coordinates of the R conformer (Brookhaven Protein Data Bank entry 1 dea) allows us to describe the geometrical changes associated with the allosteric transition as the movement of two rigid entities within each monomer. The active site, located in a deep cleft between these two rigid entities, presents a more open geometry in the T conformer than in the R conformer. CONCLUSIONS: The differences in active-site geometry are related to alterations in the substrate-binding properties associated with the allosteric transition. The rigid nature of the two mobile structural units of each monomer seems to be essential in order to explain the observed kinetics of the deaminase hexamer. The triggers for both the homotropic and heterotropic allosteric transitions are discussed and particular residues are assigned to these functions. A structural basis for an entropic term in the allosteric transition is an interesting new feature that emerges from this study. PMID- 10378273 TI - Helix swapping between two alpha/beta barrels: crystal structure of phosphoenolpyruvate mutase with bound Mg(2+)-oxalate. AB - BACKGROUND: Phosphonate compounds are important secondary metabolites in nature and, when linked to macromolecules in eukaryotes, they might play a role in cell signaling. The first obligatory step in the biosynthesis of phosphonates is the formation of a carbon-phosphorus bond by converting phosphoenolpyruvate (PEP) to phosphonopyruvate (P-pyr), a reaction that is catalyzed by PEP mutase. The PEP mutase functions as a tetramer and requires magnesium ions (Mg2+). RESULTS: The crystal structure of PEP mutase from the mollusk Mytilus edulis, bound to the inhibitor Mg(2+)-oxalate, has been determined using multiwavelength anomalous diffraction, exploiting the selenium absorption edge of a selenomethionine containing protein. The structure has been refined at 1.8 A resolution. PEP mutase adopts a modified alpha/beta barrel fold, in which the eighth alpha helix projects away from the alpha/beta barrel instead of packing against the beta sheet. A tightly associated dimer is formed, such that the two eighth helices are swapped, each packing against the beta sheet of the neighboring molecule. A dimer of dimers further associates into a tetramer. Mg(2+)-oxalate is buried close to the center of the barrel, at the C-terminal ends of the beta strands. CONCLUSIONS: The tetramer observed in the crystal is likely to be physiologically relevant. Because the Mg(2+)-oxalate is inaccessible to solvent, substrate binding and dissociation might be accompanied by conformational changes. A mechanism involving a phosphoenzyme intermediate is proposed, with Asp58 acting as the nucleophilic entity that accepts and delivers the phosphoryl group. The active-site architecture and the chemistry performed by PEP mutase are different from other alpha/beta-barrel proteins that bind pyruvate or PEP, thus the enzyme might represent a new family of alpha/beta-barrel proteins. PMID- 10378274 TI - Removal of the bridging ligand atom at the Ni-Fe active site of [NiFe] hydrogenase upon reduction with H2, as revealed by X-ray structure analysis at 1.4 A resolution. AB - BACKGROUND: The active site of [NiFe] hydrogenase, a heterodimeric protein, is suggested to be a binuclear Ni-Fe complex having three diatomic ligands to the Fe atom and three bridging ligands between the Fe and Ni atoms in the oxidized form of the enzyme. Two of the bridging ligands are thiolate sidechains of cysteinyl residues of the large subunit, but the third bridging ligand was assigned as a non-protein monatomic sulfur species in Desulfovibrio vulgaris Miyazaki F hydrogenase. RESULTS: The X-ray crystal structure of the reduced form of D. vulgaris Miyazaki F [NiFe] hydrogenase has been solved at 1.4 A resolution and refined to a crystallographic R factor of 21.8%. The overall structure is very similar to that of the oxidized form, with the exception that the third monatomic bridge observed at the Ni-Fe site in the oxidized enzyme is absent, leaving this site unoccupied in the reduced form. CONCLUSIONS: The unusual ligand structure found in the oxidized form of D. vulgaris Miyazaki F [NiFe] hydrogenase was confirmed in the reduced form of the enzyme, with the exception that the electron density assigned to the monatomic sulfur bridge had almost disappeared. On the basis of this finding, as well as the observation that H2S is liberated from the oxidized enzyme under an atmosphere of H2 in the presence of its electron carrier, it was postulated that the monatomic sulfur bridge must be removed for the enzyme to be activated. A possible mechanism for the catalytic action of the hydrogenase is proposed. PMID- 10378275 TI - The crystal structure of a reduced [NiFeSe] hydrogenase provides an image of the activated catalytic center. AB - BACKGROUND: [NiFeSe] hydrogenases are metalloenzymes that catalyze the reaction H2<-->2H+ + 2e-. They are generally heterodimeric, contain three iron-sulfur clusters in their small subunit and a nickel-iron-containing active site in their large subunit that includes a selenocysteine (SeCys) ligand. RESULTS: We report here the X-ray structure at 2.15 A resolution of the periplasmic [NiFeSe] hydrogenase from Desulfomicrobium baculatum in its reduced, active form. A comparison of active sites of the oxidized, as-prepared, Desulfovibrio gigas and the reduced D. baculatum hydrogenases shows that in the reduced enzyme the nickel iron distance is 0.4 A shorter than in the oxidized enzyme. In addition, the putative oxo ligand, detected in the as-prepared D. gigas enzyme, is absent from the D. baculatum hydrogenase. We also observe higher-than-average temperature factors for both the active site nickel-selenocysteine ligand and the neighboring Glu18 residue, suggesting that both these moieties are involved in proton transfer between the active site and the molecular surface. Other differences between [NiFeSe] and [NiFe] hydrogenases are the presence of a third [4Fe4S] cluster replacing the [3Fe4S] cluster found in the D. gigas enzyme, and a putative iron center that substitutes the magnesium ion that has already been described at the C terminus of the large subunit of two [NiFe] hydrogenases. CONCLUSIONS: The heterolytic cleavage of molecular hydrogen seems to be mediated by the nickel center and the selenocysteine residue. Beside modifying the catalytic properties of the enzyme, the selenium ligand might protect the nickel atom from oxidation. We conclude that the putative oxo ligand is a signature of inactive 'unready' [NiFe] hydrogenases. PMID- 10378276 TI - Structure of mammalian ornithine decarboxylase at 1.6 A resolution: stereochemical implications of PLP-dependent amino acid decarboxylases. AB - BACKGROUND: Pyridoxal-5'-phosphate (PLP) dependent enzymes catalyze a broad range of reactions, resulting in bond cleavage at C alpha, C beta, or C gamma carbons of D and L amino acid substrates. Ornithine decarboxylase (ODC) is a PLP dependent enzyme that controls a critical step in the biosynthesis of polyamines, small organic polycations whose controlled levels are essential for proper growth. ODC inhibition has applications for the treatment of certain cancers and parasitic ailments such as African sleeping sickness. RESULTS: The structure of truncated mouse ODC (mODC') was determined by multiple isomorphous replacement methods and refined to 1.6 A resolution. This is the first structure of a Group IV decarboxylase. The monomer contains two domains: an alpha/beta barrel that binds the cofactor, and a second domain consisting mostly of beta structure. Only the dimer is catalytically active, as the active sites are constructed of residues from both monomers. The interactions stabilizing the dimer shed light on its regulation by antizyme. The overall structure and the environment of the cofactor are compared with those of alanine racemase. CONCLUSIONS: The analysis of the mODC' structure and its comparison with alanine racemase, together with modeling studies of the external aldimine intermediate, provide insight into the stereochemical characteristics of PLP-dependent decarboxylation. The structure comparison reveals stereochemical differences with other PLP-dependent enzymes and the bacterial ODC. These characteristics may be exploited in the design of new inhibitors specific for eukaryotic and bacterial ODCs, and provide the basis for a detailed understanding of the mechanism by which these enzymes regulate reaction specificity. PMID- 10378277 TI - The crystal structure of human S-adenosylmethionine decarboxylase at 2.25 A resolution reveals a novel fold. AB - BACKGROUND: S-Adenosylmethionine decarboxylase (AdoMetDC) is a critical regulatory enzyme of the polyamine synthetic pathway, and a well-studied drug target. The AdoMetDC decarboxylation reaction depends upon a pyruvoyl cofactor generated via an intramolecular proenzyme self-cleavage reaction. Both the proenzyme-processing and substrate-decarboxylation reactions are allosterically enhanced by putrescine. Structural elucidation of this enzyme is necessary to fully interpret the existing mutational and inhibitor-binding data, and to suggest further experimental studies. RESULTS: The structure of human AdoMetDC has been determined to 2.25 A resolution using multiwavelength anomalous diffraction (MAD) phasing methods based on 22 selenium-atom positions. The quaternary structure of the mature AdoMetDC is an (alpha beta)2 dimer, where alpha and beta represent the products of the proenzyme self-cleavage reaction. The architecture of each (alpha beta) monomer is a novel four-layer alpha/beta sandwich fold, comprised of two antiparallel eight-stranded beta sheets flanked by several alpha and 3(10) helices. CONCLUSIONS: The structure and topology of AdoMetDC display internal symmetry, suggesting that this protein may be the product of an ancient gene duplication. The positions of conserved, functionally important residues suggest the location of the active site and a possible binding site for the effector molecule putrescine. PMID- 10378278 TI - Prevalence of antibodies to Coxiella burnetii (Q fever) in bulk tank milk in England and Wales. AB - In the United Kingdom, the infection of people with Coxiella burnetii, the causative agent of Q fever, is of significant public health importance and is associated with contact with dairy cattle. An ELISA was developed for the detection of IgG antibodies against C burnetii in bulk tank milk, and in a survey of randomly selected samples from dairy herds in England and Wales, 21 per cent showed serological evidence of C burnetii infection. PMID- 10378279 TI - Evaluation of ammonia measurements in dogs with two analysers for use in veterinary practice. AB - The measurement of ammonia in biological fluids is the only way to diagnose and evaluate hepatic encephalopathy, but samples for ammonia measurement cannot be stored or sent by post. Two analysers for use in veterinary practice have recently become available, the VetTest and the Blood Ammonia Checker II; the reliability of ammonia measurements in canine blood with these two analysers has been evaluated by comparing the results with a standard automated enzymatic assay. Blood samples from 39 dogs, with a range of ammonia concentrations from 5 to 589 microM, were used simultaneously in the three assays. The blood samples were placed immediately on ice, and the measurements were made in duplicate. The intra-assay coefficients of variation were 13.7 per cent for the VetTest, 4.7 per cent for the Blood Ammonia Checker, and 2.8 per cent for the enzymatic assay. The correlation coefficients over the entire range of concentrations were 0.79 between the VetTest and the enzymatic assay, and 0.98 between the Ammonia Checker and the enzymatic assay. The ammonia concentrations recorded in the enzymatic assay were divided into 12 samples within the normal range (0 to 50 microM), 18 samples with moderately increased concentrations (51 to 150 microM), and nine samples with concentrations above 150 microM. No correlation or a poor correlation was found between the results from the VetTest and those from the enzymatic assay from 0 to 50 microM (R = 0.27) and from 50 to 150 microM (R = 0.51; P = 0.05). The results from the VetTest were only reliable in samples with the highest concentrations (R = 0.93; P < 0.05). In contrast, the results from the Ammonia Checker correlated well with the results from the enzymatic assay over all the ranges: R = 0.79 (P < 0.05) from 0 to 50 microM, R = 0.86 (P < 0.05) from 50 to 150 microM, and R = 1.00 (P < 0.05) in samples exceeding 150 microM. PMID- 10378280 TI - Nationwide survey of antibodies to bovine coronavirus in bulk milk from Swedish dairy herds. AB - Bulk milk samples from 2236 dairy herds randomly selected throughout Sweden in proportion to region and herd size were analysed for antibodies to bovine coronavirus (BCV) in an ELISA. The results were expressed as optical density (OD) values and an OD > 0.04 was considered positive. Eighty-nine per cent of the samples were antibody-positive and 52 per cent had high levels of antibodies to BCV (an OD > 0.70). There were significantly higher OD values (P < 0.001) and fewer antibody-negative samples (P < 0.001) from larger herds than from smaller herds. There were also significantly higher OD values and fewer antibody-negative samples from herds in southern Sweden than from herds in northern Sweden (P < 0.001 and P < 0.001, respectively). These results indicate a higher frequency of BCV infections in larger herds and in herds in southern Sweden. PMID- 10378281 TI - Hypocalcaemia in 23 ataxic/recumbent ewes: clinical signs and likelihood ratios. AB - Twenty-three ewes in a flock of 2000 were identified as having acute onset ataxia and/or having become recumbent in late pregnancy and early lactation. The presence or absence of 15 clinical signs were recorded. Thirteen of the ewes (57 per cent) were hypocalcaemic and 10 (43 per cent) were normocalcaemic. In the hypocalcaemic group, loss of anal reflex, constipation, tachycardia, hyposensitivity, ruminal stasis, ruminal tympany, salivation and tachypnoea were recorded in 50 per cent or more of the cases. In the normocalcaemic group, tachycardia, tachypnoea and ataxia were recorded in 50 per cent or more of the cases. Constipation, ruminal stasis, salivation and hyposensitivity had likelihood ratios of 3 and above for being associated with hypocalcaemia. Ruminal stasis and hyposensitivity had the likelihood ratios of 0.10 and 0.11 respectively for not being associated with hypocalcaemia. PMID- 10378282 TI - Retrospective study of the epidemiological, clinical, haematological and biochemical findings in 109 dogs poisoned by Vipera xanthina palestinae. AB - One hundred and nine dogs were diagnosed as having been poisoned by viper (Vipera xanthina palestinae) venom between 1989 and 1996. Most of the cases occurred between April and September (86.2 per cent), with peaks in May (25.7 per cent) and July (20.2 per cent), and very few between November and February (3.6 per cent). Forty-two per cent of the dogs were poisoned in the evening (18.00 to 22.00), with a relative risk of 6.85, 17.4 per cent between 22.00 and 02.00, and 16.5 per cent between 14.00 and 18.00. The median age of the dogs was three years, and almost 80 per cent of them were from rural households. German shepherd dogs and rottweilers were over-represented (relative risk 1.98 and 1.87 respectively), and mongrel dogs and pinschers were under-represented (relative risk 0.41 and 0.53 respectively). Fifty-six per cent of the bites were on the head (excluding the mouth, lips and pinnae), 16.5 per cent on the front limbs, 9.7 per cent on the mouth and lips, 8 per cent on the hindlimbs, 4.4 per cent were submandibular and 5.4 per cent were at other sites. The main clinical signs were local swelling (98.2 per cent) and oedema (94.5 per cent), panting (45.7 per cent), tachypnoea (42.5 per cent), pain (34.9 per cent), tachycardia (29.8 per cent), lameness (25.7 per cent), and lymphadenomegaly (23.9 per cent). The mortality rate was 3.7 per cent. The most common haematological abnormalities were neutrophilia (67.6 per cent), leucocytosis (54.9 per cent), thrombocytopenia (51.9 per cent), increased haematocrit (47.6 per cent), and a left shift of neutrophils (37.8 per cent). Many biochemical abnormalities were observed, of which the most common were high activities of lactate dehydrogenase (84.6 per cent), creatine kinase (69 per cent), gamma-glutamyltransferase (40 per cent) and aspartate aminotransferase and high concentrations of globulin, phosphate and total bilirubin (33.3 per cent in each case). PMID- 10378283 TI - Activity of chlorhexidine shampoos in vitro against Staphylococcus intermedius, Pseudomonas aeruginosa and Malassezia pachydermatis. PMID- 10378284 TI - Restriction endonuclease analysis of feline herpesvirus 1 DNA isolated from wild felids. PMID- 10378285 TI - Uses of Emtryl. PMID- 10378286 TI - Abscesses in rabbits. PMID- 10378287 TI - Veterinary research--a continuing challenge for the schools. PMID- 10378288 TI - Prevalence and risk factors for feline Bordetella bronchiseptica infection. AB - A cross-sectional survey of a convenience-sample of 740 cats was undertaken to obtain an estimate of the prevalence of Bordetella bronchiseptica infection, and to identify risk factors that might predispose them to the infection. Data on individual cats and household variables, including disease status and animal contacts were obtained by questionnaire. B bronchiseptica was isolated from 82 (11 per cent) of the cats sampled. The prevalence of B bronchiseptica varied with the type of household sampled, being 19.5 per cent in rescue catteries, 9 per cent in breeding catteries, 13.5 per cent in research colonies, and 0 per cent in household pets. On the basis of a univariable analysis, 19 of 29 predictor variables were found to be significantly associated with the isolation of B bronchiseptica, including an association with cats in rescue catteries, and with cats from premises with larger numbers of animals. Separate analysis of the rescue cattery subpopulation showed a highly significant association on multivariable analysis with current respiratory disease, suggesting that different risk factors may operate in this type of environment. In the whole sample there was also strong association with cats from households containing a dog with recent respiratory tract disease. The clinical signs observed in the B bronchiseptica-positive cats included sneezing, ocular and nasal discharges and coughing, although only the association with sneezing was statistically significant. There was no significant association between the isolation of B bronchiseptica and the isolation of respiratory viruses, suggesting that in some circumstances B bronchiseptica may be able to cause disease independently. PMID- 10378289 TI - Anthelmintic treatment of dairy cows and its effect on milk production. AB - The results of more than 80 experiments on gastrointestinal parasitism and the impact of anthelmintic treatment on milk production in dairy cattle were reviewed. Abattoir surveys of culled dairy cows, faecal egg counts in milking cows, and serological tests and worm counts in cull cows in milk production studies were collated to assess the level of parasitism in dairy herds. The studies were divided into four general categories: induced infections in previously uninfected cattle; naturally infected cattle treated in mid-lactation; naturally infected cattle treated one to three times during the dry period and/or just before or just after parturition; and naturally infected cattle treated repeatedly from early lactation or given strategic treatments throughout the year. In most studies, the milk production of anthelmintic-treated cattle was compared with that of untreated controls. The anthelmintics investigated included members of the organophosphate, benzimidazole, imidazothiazole and macrocyclic lactone groups. The number of experiments in which the medicated (or uninfected) group had a higher milk yield was compared with the number of experiments in which the control (or infected) group had a higher yield. Overall, the studies demonstrated that grazing dairy cattle are likely to be infected with gastrointestinal nematode parasites, usually Ostertagia ostertagi and Cooperia species. These infections may be present as inhibited larvae, and a periparturient or spring rise is associated with their emergence. There is, at present, no reliable means of determining whether a cow or a herd may be parasitised subclinically at a level sufficient to interfere with milk production. In 70 of 87 experiments (80 per cent) there was an increase in milk production (P < 0.001) after anthelmintic treatment, with a median increase of 0.63 kg/day. In each of the four trial categories, a majority of the studies showed that anthelmintic treatment increased milk production. The yield of milk fat by the medicated cows was greater than by the controls in 26 of the 35 experiments in which that variable was studied (P < 0.01). PMID- 10378290 TI - Evidence of Brucella infection in marine mammals in the North Atlantic Ocean. AB - Between 1983 and 1996 a total of 1386 samples of serum were taken from four species of seal and three species of whale in the waters west of Iceland, the area of pack-ice north-west of Jan Mayen, the northern coast of Norway and the Kola Peninsula, the waters west of Svalbard, and the Barents Sea; they were tested for the presence of anti-Brucella antibodies with an indirect ELISA (protein G conjugate). The positive sera were re-tested with classical brucellosis serological tests, such as the serum agglutination test, the EDTA modified serum agglutination test, the Rose Bengal test, and the complement fixation test, as well as an anti-complement ELISA. Anti-Brucella antibodies were detected in all the species investigated, except for the bearded seal (Erignathus barbatus), with the following prevalences: hooded seals (Cystophora cristata) 35 per cent; harp seals (Phoca groenlandica) 2 per cent; ringed seals (Phoca hispida) 10 per cent; minke whales (Balaenoptera acutorostrata) 8 per cent; fin whales (Balaenoptera physalus) 11 per cent; and sei whales (Balaenoptera borealis) 14 per cent. An isolate belonging to the genus Brucella was obtained from the liver and spleen of one of the seropositive minke whales. The findings suggest that antibodies against the surface lipopolysaccharide of Brucella species are widely distributed among marine mammals in the North Atlantic Ocean. PMID- 10378291 TI - Cutaneous papillomavirus infection in a harbour porpoise (Phocoena phocoena) from the North Sea. PMID- 10378292 TI - Bovine tuberculosis. PMID- 10378293 TI - Bovine tuberculosis. PMID- 10378297 TI - [Regulation of intracellular CoA pool as one of approaches to correction of metabolic disorders]. AB - The problems of biosynthesis of coenzyme A, its transport into the mitochondria, and compartmentalization in mammalian cells have been reviewed. Co A pool structure in liver cells and the in myocardium under different pathobiochemical conditionsis discussed. Experimental data which have now been accumulated can be used as a basis for correction of the metabolic disturbances of different etiology. PMID- 10378298 TI - [Bioavailability of carotenoids]. AB - The review analysis of the gradual process of the assimilation of carotenoids in the living organism depending upon various factors of both external and internal environment is given. Elucidation of in time mechanisms of uptake of carotenoids can be of both theoretical and practical significance. It will allow to modify the composition of carotenoids-based drugs, dietary additives and balanced diets, to work out the recommendations on its dosage regimen for different groups of people. PMID- 10378299 TI - [Metabolic adaptation to alcohol in rats with different preference of ethanol over water]. AB - Recent studies have shown that the phenomenon of ethanol preference by animals and of alcohol consumption by humans may be related to the intensity of its metabolism in the body and depend on the activities of the ethanol and aldehyde metabolizing systems which are potential regulators of the acetaldehyde level in the cell. The special features of adaptative reactions of this system (alcohol dehydrogenase, microsomal ethanol oxidizing system, catalase, aldehyde dehydrogenase) were examined in rats, differing by the preference to water or ethanol (5%, 10%, 15%) under condition of a long contact with alcohol. PMID- 10378300 TI - [Acidity and interaction with superoxide anion radical of echinochrome and its structural analogs]. AB - Weak acid properties, autoxidation and interaction of natural polyhydroxy1,4 naphthoquinones (PHNQ) with superoxide anion-radical (O2-.) were studied by methods of potentiometric titration, polarography, and UV- and visible spectrophotometry. Sea urchin pigments 3-acetyl-2,6,7-trihydroxynaphthazarin (spinochrome C), 2,3,6,7-trihydroxynaphthazarin (spinochrome D), 2,3,6,7 trihydroxynaphthazarin (spinochrome E), 6-ethyl-2,3,7-trihydroxynaphthazarin (echinochrome A), synthetic 2,3-dihydroxy-6,7-dimethylnaphthazarin and 6-ethyl 2,3,7-trimethoxynaphthazarin (trimethoxyechinochrome A) were tested. Determined dissociation constants (pKi) were in the range of pH 5.3-8.5 (40% ethanol solvent). PHNQ autoxidation observrd in basic pH were inhibited by superoxide dismutase. Xanthine and xanthine oxidase was applied for O2-. generation. Interaction with O2-. led to sufficient time-dependent changing in spectra of echinochrome A, spinochromes D and E. There was weak O2-. influence on spinochrome C spectrum and no changing in trimethoxyechinochrome A spectrum. The spectra, that were transforming during time of reaction, contained pronounced isobestic point. It means formation the single reaction product. We proposed formation of 1,2,3,4-tetraketones from 2,3,5,8-tetrahydroxy-1,4-naphthoquinones (echinochrome A, spinochromes D and E) due to O2-.-induced oxidation of their OH groups in 2 and 3 positions. Reaction constants were determined by competition method using nitro blue tetrazolium (NBT). The reaction constants were about 10(4)-10(5) M-1s-1. They were decreased in the order: echinochrome A > spinochrome D > spinochrome C > NBT > trimethoxyechinochrome A. Thus, we concluded that some of the natural PGNQ, containing hydroxyl groups in 2nd and 3rd positions, could operate as powerful superoxide anion-radical scavengers. PMID- 10378301 TI - [Lipid parameters of the skin, cerebellum, and medulla oblongata during immersion stress in rats]. AB - The influence of short-form water immersion stress of rats on lipids in the skin, the cerebellum and the medulla oblongata was studied. The level of total lipids and absolute and relative contents of the main lipid fractions (phospholipids, nonesterified cholesterol, free fatty acids, triglycerides, and cholesterol esters) were measured. Stress induced delayed changes of the lipid component of the skin. The first significant changes of lipid fractions were only observed 20 h later after the stress procedure. These changes were retained (being at nearly constant levels) till the end of the second day. The decrease in contents of total lipids and esterified cholesterol was revealed in the cerebellum of stressed rats (in comparison to these levels in control rats). These results suggest the involvement of cholesterol metabolic system in the stress reaction. The content of total lipids decreased also in the medulla oblongata. However, levels of the main lipid fractions changed differently. The content of diglycerides increased and the content of cholesterol decreased. The data obtained suggest that degradation of triglycerides is the principle pathway of metabolic conversions of lipids. Free fatty acids formed during these processes are probably involved in the synthesis of phospholipids and cholesterol esters. The data indicate absolutely different mechanisms of interrelations between individual lipid fractions in the brain regions studied. Various roles of the brain structures in the stress response of the body may account for the differences revealed. PMID- 10378302 TI - [Specific binding and uptake of peptide ligand modified liposomes by PC12 cells]. AB - Possible employment of cell-specific peptide for the specific adsorption and uptake by cells. It was shown, that apoprotein E 139-158 peptide increases liposomal binding followed by receptor-mediated endocytosis by cells PC12. PMID- 10378303 TI - [Lipid peroxidation during enteral correction of experimental blood loss]. AB - Time course of lipid peroxidation and the state of antioxidant system of dog blood in an experimental haemorrhage were studied. Increases of conjugated dienes, malonic dialdehyde, Shiff's bases contents, lipid peroxidation degree were revealed. Enteral correction of the haemorrhage by the glucose-aminoacid saline enriched by mafusol promotes a decrease of lipid peroxidation. PMID- 10378304 TI - [ATPase activity in neurons and neuroglia during convulsions induced by picrotoxin]. AB - Na+,K(+)-ATPase activity was studied in neurones and neuroglia under conditions of convulsions caused by picrotoxin administration. Picrotoxin is a stimulant which causes convulsions by suppression of presynaptic inhibition. Na+,K(+) ATPase activity in neuroglia was increased in convulsion, bat was not altered in neurones. It is possible, that glial Na+, K(+)-ATPase activity plays the role of neuronal ion's flows regulator and neuronal protector from convulsions. In recovery period ATPase activities in neuronal and neuroglial cells run up to control. PMID- 10378305 TI - [Metabolism of vitamins B1 and B2 during phenylketonuria]. AB - Vitamin level in blood plasma and erythrocytes and a rate of urinary excretion of vitamin metabolites were analysed in children suffering from phenylketonuria. It has been shown that vitamin B1 metabolism and therefore the criteria of the adequate sufficiency with this vitamin does not differ from those for healthy people. Increased riboflavin urinary excretion under its decreased plasma and erythrocyte levels has been demonstrated for PKU children. In consequence of this the indexes of sufficiency significantly differ from those of the healthy adequately supplied with this vitamin children and are equal to 4 ng of riboflavin per 1 ml of blood plasma and its urinary excretion more than 50 mg/h. The necessity for the redetermination of vitamin B2 diet optimal content under this disease and its biochemical validation are discussed. PMID- 10378306 TI - [Fibrinolytic activity of the urine during chronic glomerulonephritis and amyloidosis]. AB - Correlative interconnections between plasminogen activator (PA) activity (fibrin plate method) and level of urokinase antigen (Ag UAP) and tissue PA antigen (Ag TAP) in urine and blood (ELISA) were studied in 60 patients with chronic glomerulonephritis (CGN) and 38 patients with amyloidosis. The high degree of positive correlation between blood and urine initial PA activity and Ag UAP content was found. This suggests the possible leading role of UAP in formation of the basal fluctuations of fibrinolytic activity in blood and urine. High degree of correlation--r = +0.84 and p < 0.001--was found between blood Ag UAP and urine Ag TAP in amyloidosis only. The functional protein loading probe revealed great importance of high urine and blood AP activity in realizing of ultrafiltration renal process--in CGN and amyloidosis. PMID- 10378307 TI - [Comparative and quantitative analysis of proteins in the urine peritoneal fluid, separated by gel electrophoresis and stained with silver]. AB - The method for the silver staining of proteins fractionated with SDS electrophoresis is described as a new approach for the complex analysis of biological fluids allowing to obtain the valuable diagnostic information and to determine quantitatively individual proteins in urine and peritoneal fluid, in particular in nephropathologies. PMID- 10378308 TI - [Development of a two-site immunoassay using polyclonal antibodies for determination of lactoferrin in human blood]. AB - The two-site enzyme-linked immunosorbent assay (ELISA) for lactoferrin using polyclonal antibodies to spatially distant epitopes has been developed. The assay sensitivity defined as minimal detectable lactoferrin concentration for p = 0.05 is 0.5 ng/ml. Accuracy of the assay (variance coefficient) is 7% within the clinical range of antigen concentrations. Human albumin, hemoglobin, and transferrin in concentrations up to 5 mg/ml practically do not interfere with the measurement. Sera of healthy donors and viral hepatitis patients were investigated using the two-site ELISA. The lactoferrin content in 44 donors' sera was 130 +/- 40 ng/ml (medium +/- standard deviation). A study of the serum specimens of 95 patients with hepatitis A, B, and C showed significant increase in serum lactoferrin concentration: 850 +/- 420, 780 +/- 580, and 680 +/- 500 ng/ml respectively. The assay showed good characteristics and may be recommended for lactoferrin measurement in patients' sera. PMID- 10378309 TI - [Computer modeling in the study of mechanism of catalytic activity and the structure of active site of glutamine(asparagine)ase. I. Pharmacophore models of glutamine(asparagine)ase substrates]. AB - Glutamine(asparagine)ase catalyses desamidation of both L-glutamine and L asparagine, and their D-isomers. In this study the two-pharmacophore models of main enzyme substrates and their hydrolysed analogues were design. The received models reflect two stage of substrate interaction with the enzyme active site. These models allow to explain the wide substrate specificity of glutamine(asparagine)ase. PMID- 10378310 TI - [Two hundred years ago: the first smallpox vaccinations in Vienna]. AB - The first successful smallpox vaccination with cowpox lymph outside of England was carried out in Vienna--only ten months after the publication of Edward Jenner's book "An Inquiry into the Causes and Effects of the Variolae Vaccinae, a Disease ... known by the Name of the Cow Pox", and only a little more than three months after the first vaccinations in London: Two hundred years ago, on 30 April 1799, the medical service chief of Lower Austria, Dr. Paskal Joseph Ferro, born in Bonn, vaccinated his three children with vaccine which had come in a letter from London. Subsequently the vaccination was introduced in Austria. Before 1800 effective prophylactic immunizations against smallpox were carried out, apart from England, only in Vienna and environs. The first efficient vaccine to reach India also came from Vienna. PMID- 10378311 TI - Hyperlipidemia and renal disease: the use of animal models in understanding pathophysiology and approaches to treatment. AB - Hyperlipidemia accelerates the progression of human renal disease, and its control is becoming an important component of therapy for patients with renal failure. The biologic basis for these observations remains poorly understood. This review summarizes recent data from animal models which show how lipoproteins interact with cells directly to cause renal injury. Data are presented from recent studies on the obese Zucker rat, a metabolic model of hyperlipidemia, obesity, and glomerular sclerosis, showing that triglyceride-containing lipoproteins may mediate glomerular injury. The biochemical basis for therapy is also discussed, including actions of lipid-lowering agents apart from their effect on serum lipids, and the use of polyunsaturated fatty acids as dietary therapy. Animal models are a crucial tool for the further elucidation of mechanisms of lipoprotein-mediated glomerular injury. PMID- 10378312 TI - [Trends in molecular diagnosis]. AB - The number of characterized monogenic and polygenic diseases is rising each year. In consequence, molecular diagnostics is faced with an ever increasing number of patient samples and with more and more heterogeneous genetic defects. The fusion of microelectronics and molecular biology has created a new technology (microelectronic miniaturization), which provides a rapid, efficient, and cost effective tool in molecular diagnostics at a high-sample throughput. The biochip has recently been selected as one of the ten scientific highlights in the year 1998. The application of microelectronics ranges from the polymerase chain reaction (PCR), nucleotide sequence analysis via DNA-chips or capillary electrophoresis-chips to gene expression analysis. These microchips are suited for integration into fully automated systems, thus providing the basis for automation of molecular diagnostics. The present article summarizes important trends in molecular diagnostics and provides a glimpse on future technologies. PMID- 10378313 TI - Occurrence and relevance of postprandial hypotension in patients with essential hypertension. AB - The aim of this study was to determine the occurrence of postprandial hypotension (PPH) in patients with arterial hypertension, identify its risk factors and evaluate the importance of postprandial blood pressure reduction in relation to the management of hypertension. Forty-nine patients (23 male; 26 female; mean age 65.6 +/- 12 years) with diagnosed hypertension underwent measurement of blood pressure and pulse rate before intake of a standardised breakfast (1821 kJ) and at 15-minute intervals until 1 hour thereafter. The orthostatic test for detection of orthostatic hypotension was performed before the ingestion of food. PPH was detected in 22 patients (45%) with arterial hypertension. Patients treated with diuretics had significantly greater postprandial reductions in blood pressure compared to those who received no diuretic treatment. Levels of premeal systolic blood pressure, age, orthostatic hypotension, history of syncope cardiovascular disease or stroke were not associated with a more severe decline in postprandial blood pressure. Maximal reductions in blood pressure were recorded approximately 33 +/- 15 minutes after ingestion of food. Therefore, recent intake of food should be taken into account in the evaluation of hypertension and the effect of antihypertensive drugs when blood pressure is measured within one hour after a meal. PMID- 10378314 TI - Serum uric acid concentration and thyroid-stimulating-hormone (TSH): results of screening for hyperuricaemia in 2359 consecutive patients with various degrees of thyroid dysfunction. AB - Serum uric acid concentration (sUA) and hyperthyroidism have been reported to positively correlate with each other. Furthermore, epidemiological data indicate that uric acid may be an independent risk factor for hypertension-associated morbidity and mortality. To evaluate whether screening for hyperuricaemia might be worthwhile in patients with hyperthyroidism we determined serum concentrations of uric acid in 2359 consecutive patients (1939 female, 420 male; age: 48 +/- 17 years, mean +/- SD) with various degrees of thyroid dysfunction (hyperthyroidism: n = 242; subclinical hyperthyroidism: n = 143, hypothyroidism: n = 71, subclinical hypothyroidism: n = 212) and in 1688 euthyroid subjects. No association (r = 0.03) between sUA and total T4/TSH was detected. The significant difference (p < 0.05) in serum uric acid between hyperthyroid (4.8 +/- 1.32 mg/dl) and euthyroid (4.5 +/- 1.32 mg/dl) patients was of no clinical significance. We conclude that routine determination of sUA in hyperthyroid patients is not warranted. PMID- 10378315 TI - [Primary lymphoma of the thyroid with contralateral recurrence--case report]. AB - Primary lymphomas located in the thyroid gland are rare clinical findings. The therapy and the subsequent monitoring of the disease continue to be a subject of debate. We present the case of a 63-year-old female patient in whom a hemi thyroidectomy was performed because of a growing goiter. Histological examination of the excised tissue revealed a large-cell B-lymphocytic lymphoma which extended to neighboring lymph nodes. The patient received chemotherapy which led to remission of disease for two years. The disease re-occurred in the remaining thyroid lobe. Subsequently, the patient was treated with involved-field radiation therapy which lead to long term remission. We conclude that total thyroidectomy should be considered in the treatment of lymphomas located in the thyroid gland. PMID- 10378316 TI - ["Ambulatory" therapy of deep venous thrombosis of the leg--definition]. AB - The central problem in connection with "ambulatory treatment" of deep vein thrombosis is the degree of mobility. In future trials it will be essential not only to register carefully anticoagulant therapy but also to define and to measure walking activities and adjuvant compression. PMID- 10378317 TI - [The death of Thomas Mann: consequence of erroneous angiologic diagnosis?]. AB - Soon after his 80th birthday in June 1955 celebrated in Kilchberg near Zurich Thomas Mann developed a swollen and painful left leg during his vacation on the seashore of Holland. The diagnosis of "thrombophlebitis" was made by Mulders from Leiden and confirmed by Wilhelm Loffler in Zurich. To the surprise of the treating doctors an irreversible shock occurred, when the famous poet and Nobel laureate was improving. The pathological diagnosis (Christoph Hedinger) was perforation of an iliac artery aneurysm on the left side with extensive retroperitoneal hematoma, compression and thrombosis of the iliac vein. Because of lacking apparative techniques correct intra vitam diagnosis was not yet possible in 1955, and life saving vascular surgery was available only shortly later. It is described how Thomas Mann himself experienced his last illness. His astonishing medical knowledge is illustrated by examples dealing with vascular medicine. A summary of his last year of life is given. PMID- 10378318 TI - [Pathophysiology of immobilization]. AB - Deep venous thrombosis is initiated primarily in the pockets of the valves of the veins of the lower limbs and the main pelvic veins. In addition to a genetic predisposition there are several acquired conditions associated with deep venous thrombosis such as major surgery, trauma, cancer, pregnancy, and immobilization. While major diseases as well as hormonal changes have been shown to cause changes in blood coagulation and fibrinolytic factors, the impact of immobilization per se is much less clear. It has been shown that in the absence of intermittent pulsatile flow the blood within the valve pockets became rapidly hypoxic when undisturbed during streamline flow (i.e. when "static"). Hypoxia on the other hand has been shown to cause procoagulatory changes of the vascular endothelium (e.g. production of platelet activating factor [PAF], expression of tissue factor), adhesion and activation of leukocytes and expression of tissue factor on their surface, as well as the activation of platelets. Together with reduced removal and/or inactivation of active clotting factors these mechanisms might contribute to the development of deep venous thrombosis during immobilization. PMID- 10378319 TI - [Predisposition and initiation of venous thrombosis]. AB - The causes of thromboembolic disease are complex. In addition to acquired risks, thrombophilic diatheses in particular now play an important role. The combination of predisposing factors increases the risk of thrombosis. However, a precise prognosis cannot yet be made for an individual patient. PMID- 10378320 TI - [Standard heparin versus low-molecular-weight heparin]. AB - Deep vein thromboses of the legs, which have been treated with low molecular weight heparin (LMWH), show significantly less thromboembolic recurrencies, less extension of the thrombus, fewer bleedings and a lower mortality than standard heparin (UFH). The differences are, however, small. LMWH is considerably more expensive. The advantage of LMWH lies in the possibility that it can be subcutaneously injected and thus given on an outpatient basis. This makes the treatment much easier and reduces the treatment costs by 50%. PMID- 10378321 TI - [Therapy of thromboses with low-molecular-weight heparins]. AB - Low-molecular-weight heparins, nowadays already widely used for the prevention of thromboembolism, have now also become available for the treatment of deep-vein thrombosis. This article should serve to explain the rationale for this development and to demonstrate the clinically relevant advantages of the use of low-molecular-weight heparins. After briefly describing the characteristic properties of heparins the most relevant studies comparing the use of low molecular-weight heparin versus unfractionated heparin for the treatment of thromboembolism are discussed. In conclusion, clinical trials suggest that low molecular-weight heparins given subcutaneously can replace the hitherto standard intravenous application of unfractionated heparin in the initial treatment of deep-vein thrombosis, granting equal or even better efficacy and potentially lower rates of adverse side effects. Furthermore, the simplicity of this therapeutic regime allows for treatment of patients at home, thus offering patients' mobility and also reducing the cost of treatment. PMID- 10378322 TI - Outpatient treatment of deep vein thrombosis with LMWH. AB - Low-molecular-weight heparin therapy is safe and effective in the treatment of patients with acute deep vein thrombosis. These compounds are administered subcutaneously, without a need for laboratory control. This has made possible to treat patients with venous thrombosis outside the hospital. Two large studies, Tasman and a Canadian study, have demonstrated the efficacy and safety of low molecular-weight heparin in this setting. Furthermore, there is some evidence that the safety, efficacy and feasibility is good in daily routine practice. Now, the task is to develop guidelines for widespread application of these findings to daily clinical practice. However, successful home treatment will require intensive patient education as well as an extensive infrastructure of supportive nursing and physician services. Careful patient follow-up will be crucially important. PMID- 10378323 TI - [Thrombophlebitis: bed rest or walking exercise?]. AB - It is a common tradition to admit patients with deep vein thrombosis (DVT) to the hospital and put them to bed for several days because of fear from pulmonary embolism, even if they are mobile. Between May 1994 and December 1997 929 patients were admitted to our department who were treated by subcutaneous injections of low-molecular-weight heparin (mainly 200 IU dalteparin per kilogram body-weight per 24 hours), got firm compression bandages and were encouraged to walk as much as possible. On admission DVT propagated into the pelvis in 268 patients, into the thigh in 480 and below the popliteal level in 181 patients. V/Q-lung scans were performed at baseline and repeated after 10 days on average. In these three groups primary pulmonary embolism was diagnosed in 49.4%, 50% and 34% respectively, new emboli after 10 days were found in 6.1%, 5.7% and 3.9%. Only one third of the patients with embolism on admission and 5 from 50 patients who developed new emboli showed some dyspnoea. 12 patients died and underwent autopsy, 3 fatal events were caused by pulmonary embolism. With out management the incidence of thromboembolic complications is statistically significantly lower than data from the literature. Preliminary results from an ongoing randomised trial comparing bed-rest, compression bandages and compression stockings in the acute phase of proximal DVT demonstrate faster improvement of swelling and of pain in the compression-groups. Low-molecular-weight heparin has greatly facilitated therapy of DVT since effective anticoagulation can be obtained by subcutaneous injections of fixed doses without the need of laboratory monitoring. For the future development of conservative management mechanical prophylaxis of thrombus extension by acceleration of venous flow using leg compression and walking will probably become as important as exact anticoagulation. PMID- 10378324 TI - [Value of compression therapy in treatment of deep venous thrombosis]. AB - Therapy in the acute phase of deep vein thrombosis aims to prevent the longitudinal growth of the thrombus as well as the life-threatening complication of pulmonary embolism. Compression therapy is part of all revascularisative and conservative treatment strategies, but hardly mentioned in most of the literature. Clinical efficacy of a sufficient compression therapy in patients with acute deep vein thrombosis is marked by a quick ease of subjective complaints as well as a reduction of the edema. The long-term risk of deep vein thrombosis, the postthrombotic syndrome, is avoidable by the use of compression therapy. Clinical studies about the effectiveness in relation to the length of the thrombus and the incidence of pulmonary embolism are missing as well as studies about the selection of the best compression material. Recently compression pressures were experimentally measured between long and short stretch bandages and the skin in order to select bandage material. It can be concluded that long stretch bandages can be used in the treatment of acute phlebothrombosis in mobile out-patients. Short stretch bandages should be used in immobile patients in order to avoid high pressures between the skin and the bandages. Compression stockings (Class III) represent an equivalent alternative to compression bandages after the resolution of edema. PMID- 10378325 TI - [Modern thrombosis management, also in Germany]. AB - Foreign phlebologists from Austria, Sweden and Switzerland have been demonstrating successfully for years that alternative approaches to the traditional treatment of DVT exist. At the outpatient clinic for vascular diseases in Essen, 520 patients with acute deep vein thrombosis have been treated since January 1996, approximately 85% of which on an outpatient basis. The results are quite encouraging. No complications had to be faced. 188 patients were highly satisfied with the fact that they could stay at home, could return to work considerably sooner than after being referred to hospital, not to speak of an absolutely convincing reduction of therapeutical costs. On the other hand, the therapeutical concept demands high flexibility, adequate equipment and an effective communication network between patient, specialist, family doctor and clinic, and should be used by experienced vascular specialists only. PMID- 10378327 TI - The effect of the duration of anticoagulation and other risk factors on the recurrence of venous thromboembolisms. Duration of Anticoagulation Study Group. AB - BACKGROUND: In order to find the optimal balance of anticoagulation, avoiding recurrent events as well as major hemorrhage, it is important to know the impact of different risk factors over time. METHODS: Based on a randomized, multicenter trial on different durations of anticoagulation after a first episode of venous thromboembolism, the probability of recurrence over 6 years of follow-up was analyzed. The contribution of the site of the thrombus, the nature of the triggering risk factor and the presence of cardiolipin antibodies to the risk of recurrence was also calculated. RESULTS: There is a cumulative risk of recurrence of 4 to 5% per year, which continues through year 6 after the index event, independent of the initial duration of anticoagulation. Deep vein thrombosis proximal to the knee joint or symptomatic pulmonary embolism confers a risk of recurrence that is higher than of distal thrombosis but equal to the risk when the triggering factor is permanent or unknown. The combination of proximal deep vein thrombosis or symptomatic pulmonary embolism and permanent/unknown triggering factor increases the risk, and the presence of cardiolipin antibodies generates an additional risk in all subgroups. CONCLUSION: It is necessary to tailor the duration of anticoagulation individually, according to the presence of different risk factors. PMID- 10378328 TI - [Monitoring blood coagulation]. AB - Standard heparin in therapeutic doses has to be monitored by the activated partial thromboplastin time. There is no need for monitoring of treatment or prophylaxis with low molecular weight heparins. Only specific clinical situations, like renal insufficiency, long-term treatment, pregnancy, high risk of bleeding or thrombosis, small children and an extremely low or high body weight demand determination of anti-factor Xa activities. Monitoring of oral anticoagulant treatment should be done by determination of prothrombin time, values should be given in International Normalized Ratio (INR). It has been shown that monitoring by specialized centers and most probably self monitoring at home by the patient himself are able to optimize treatment. PMID- 10378329 TI - [Incidence of pulmonary embolism in venous thrombosis]. AB - The frequency of pulmonary embolism in patients with deep vein thrombosis can be assessed by pathological-anatomical and by nuclear medical studies. The frequency of deep vein thrombosis in autopsies ranges from 23.7% to 62%, by inclusion of microscopic thrombi the frequency increases to 72%. In most cases the localisation of the venous thrombosis is bilateral. In cases of venous thrombosis the frequency of pulmonary embolism is 52 to 79.4%, if microscopic thrombi are included, the number is 87.8%. 7.8% to 78.9% of all pulmonary emboli are considered as cause of death or severely contributing to death. The fatal embolisms originate preferentially from iliofemoral thrombosis. About 1/10 of all emboli originate from V. cava sup. and the right heart. In a small number of pulmonary emboli the origin could not be detected. The rate of correct intra vitam diagnosis is low, not more than 11 to 25% of all pathological-anatomical proven emboli had a correct diagnosis during life. In fatal pulmonary embolism the correct clinical diagnosis was made in 1/3. In nuclear medicine studies pulmonary embolisms are searched for from the clinical suspicion or the diagnosis of deep vein thrombosis with the perfusion-/ventilation- or inhalation scintigraphy. Patients with deep vein thromboses showed in 38 to 57.9% pulmonary embolism. In 80% of all pulmonary embolism multiple perfusion defects (2 to 9 perfusion defects) were detected, the lesions were evenly distributed in both lungs. The frequency of pulmonary embolism in calf vein thrombosis was 46%, in leg vein thrombosis 67% and reached 77% if the pelvic veins were involved. It is remarkable that the majority of all pulmonary emboli (46.3% to 100%) showed no clinical symptoms. The knowledge about the high frequency of pulmonary embolism in patients with deep vein thrombosis can improve the diagnosis of pulmonary embolism. For the diagnostic process of pulmonary embolism the presence of acute deep vein thrombosis increases the pre-test probability (prevalence of more than 50%). Every positive test for pulmonary embolism will gain a very high post-test probability according to Bayes' theorem. PMID- 10378330 TI - [Clinical aspects and diagnosis of pulmonary thromboembolism]. AB - The symptomatology of PTE is good for the suspicion of PTE only. Symptoms are mainly based on dyspnoea (respiratory rate) and pain in the chest. Clinical diagnostic procedures, on the other hand, are backed up by evaluation of specific risk factors (previous thrombosis, post-operative state, immobilisation) and search for deep venous thrombosis (DVT). Blood gas analysis is very sensitive to unravel (latent) respiratory failure, along with established routine measures (blood pressure, ECG, chest-film) and additional echocardiography. Most important in our experience is a perfusion scan at the earliest opportunity. Spiral-CT angiography is indicated in special cases only. We looked at 115 consecutive patients with suspected PTE and found close correlations between risk profiles of thrombosis, pathological BGA and high probability perfusion scans. DVT was detected in 50% only. Positive predictive values for high risk and pathological BGA were 86 and 92%, respectively. An algorithm for diagnostical/therapeutical strategies is presented. The early application of an heparin-bolus is stressed. PMID- 10378332 TI - [Stemmer's sign--possibilities and limits of clinical diagnosis of lymphedema]. AB - The sign of the thickened cutaneous fold of the second toe is typical for the early and differential diagnosis of a primary ascending lymphedema without false positive findings. It appears in the late stages of the descending lymphedema. PMID- 10378331 TI - [Prospective 12-year follow-up study of clinical and hemodynamic sequelae of deep venous thromboses in patients with low risk (Zurich Study)]. AB - No prospective study of the long-term sequelae of more than 10 years after acute deep vein thrombosis exists so far. Therefore, 28 patients with DVT were included in a prospective study to evaluate the natural history of postthrombotic syndrome. Clinical and hemodynamic examinations were performed at the time of admission; after 3, 6 and 12 months; after the 2nd, 3rd, 4th, 5th; and finally after the 12th year. All patients received unfractionated heparin initially and oral anticoagulants subsequently. After 12 years, 64% of the patients exhibited normal findings. Mild skin changes were found in 28%, marked trophic changes in 5%, and only 1 venous ulcer occurred. Regular use of compression stockings was reported by 54% of the patients with multilevel disease. Although mean maximum venous outflow was significantly reduced from the acute event to 2 years later (p < 0.003) compared with the contralateral leg, a significant (p < 0.05) improvement was observed 6 months later. Recanalization of calf vein thrombosis was detected by Doppler sonography after 3 months. 64% of the multilevel thromboses were recanalized completely or in part after 1 year; in 69%, valvular incompetence was found. In conclusion, in contrast to earlier reports, this prospective study up to 12 years after DVT demonstrates a low incidence of PTS by administration of initially unfractionated heparin, oral anticoagulation and compression therapy. However, the adverse clinical event rate (mortality 14%) and recurrency rate of 24% show that the prognosis after DVT does not appear favorable even in low-risk patients. PMID- 10378333 TI - [Isotope lymphography--possibilities and limits in evaluation of lymph transport]. AB - Quantitative isotopic lymphscintigraphy is based on a combination of transmission and emission scintigraphy correcting the different depths of lymph nodes. This method allows an exact estimation of lymph-transport in legs during standardised stress. The depth-corrected uptake in the lymph nodes expressed in percentage of the injected dose (D%) reflects the lymph-transport. After s.c. injection of 1 mCi of 99 m Tc-microcolloid (Nanocoll) the prefascial lymph-transport and after intramuscular (i.m.) injection the subfascial lymph-transport can be assessed. The stress consists of 15 minutes walking on a horizontal treadmill at a speed of 3.2 km/h. After s.c. injection all types of lymphedema can be diagnosed (average uptake in lymphedema 2.0 +/- 2.5 D%, in normal legs 14.3 +/- 4.2 D%; p < 0.001). The subfascial transport is much lower, only 7.7% of the prefascial transport. In healthy legs the uptake after i.m. injections is 1.1 +/- 0.8 D%, in postthrombotic syndrome the uptake decreases to 0.2 +/- 0.16 D% (p < 0.02). Lymphscintigraphy after i.c. injection of the colloid reveals contradictory results. The uptake is very low only in lymphedema with involvement of the whole leg. In distal and in secondary lymphedema the uptake is near the normal range. The clearance rate of the tracer from the depot is not reliable for diagnosing lymphedema. Information may be gained concerning the following points: quantitative measurement of lymph transport, routine diagnosis of lymphedema, follow-up in lymphedema, assessment of pre- and subfascial lymph-transport in patients with venous diseases, angiodysplasias etc., documentation of therapeutic effects. PMID- 10378334 TI - [Indirect lymphography--possibilities and limits of roentgen diagnosis of the lymphatic system]. AB - Indirect lymphangiography enables to opacify peripheral lymph-collectors. In case of lymph valve insufficiency and dermal back flow also small skin lymphatics can be visualized. Indirect lymphangiography showed various new lymphatic patterns in different kinds of leg oedema. Specificity of 89% and sensitivity of 97% recommends the method as a good diagnostic tool for diagnosis of lymphoedema in a suspected clinical situation. Indirect lymphangiography is not able to opacify proximal or central lymphcollectors and lymph nodes. PMID- 10378335 TI - [Artificial lymphedema from the clinical viewpoint]. AB - Factitious lymphoedema should be taken into account in patients with recurrent oedema of a limb, characterized by a distinct border, pain and an onset discordant to the accused event. PMID- 10378336 TI - [Current status of diagnosis of lymphedema--a comprehensive Austrian survey of hospitals and patients]. AB - By means of two questionnaires sent out to all Austrian hospitals and to lymphedema patients data about diagnostic tools have been collected. The geographic distribution as well as the number of different diagnostic tools were highlighted. PMID- 10378337 TI - [Ambulatory drainage of lymphedema--possibilities and limits of ambulatory management of patients with lymphedema]. AB - Manual lymph drainage on an outpatient basis is safe, effective and produces low costs. PMID- 10378338 TI - [Revolving door patient in lymphology--possibilities and limits of therapy and patient motivation during inpatient and ambulatory conditions]. AB - Patients suffering from chronic diseases necessitate a continuous treatment and self-care. Independent from its etiology the method of choice of the treatment of chronic lymphedemas of the limbs is "Complex Decongestive Physiotherapy" (CDP). CDP is carried out according to the requirements either as an outpatient- or an inpatient-treatment. In some cases CDP has to be applied not in its usual, regular form but in a modified manner. The lecture deals with problematic cases and with the limits of our therapeutic possibilities. PMID- 10378339 TI - [Post-therapeutic lymphedema of the arm--possibilities and limits of diagnosis and therapy]. AB - The post-therapeutical secondary arm lymphedema is the most frequent complication after a curatively treated cancer of mamma. For the diagnosis and therapy the knowledge of physiology and pathophysiology of lymphedema and of specific anatomy are necessary. The diagnosis facilities are essentially limited to a basic diagnosis (anamnesis, inspection, palpation, sonography, functional-diagnosis). Specific apparative diagnostics like lab, sonography, CT, MRI and PTE have to be applied especially at an early stage of the secondary arm lymphedema for the differential diagnosis between the secondary malign and secondary benign arm lymphedema. Specific apparative examinations like lymphscintigraphy and lymphography are limited and solely indicated for special questions. As a therapy possibility of the secondary arm-lymph edema, a conservative therapy, that is, the complex two-stage-decongestive physiotherapy (CDP) is recommended as first choice. Surgical therapies such as autologous lympho-lymphostatic anastomoses and lymphovenous anastomoses are only recommended in selected individual cases. The secondary malignant arm lymphedema must be primarily treated oncologically; lymphological therapy measures have to be postponed. Diagnosis and therapy are limited through lymphological incompetence and insufficient patient compliance. In this respect the provision of financial resources through National Health policy ist regarded as utterly important. PMID- 10378340 TI - Possibilities and restriction of isotopic lymphography for the assessment of therapeutic effects in lymphedema. AB - Whatever the results are, we must keep in mind that the method used must be safe, simple, reproductive, quite physiological. The RNL with distal registration during 40 min give more precise and reliable informations particularly in transient or permanent lymphatic dysfunctions. This should be the protocol used for the assessment of the lymphatic function and the treatment efficacy. The RNL with nodal uptake registration is totally related to the lymphatic transport from the extremities and the exercise applied. When performed by well trained physicians concerned with the lymphatic physiology and informed of all factors interfering with the results, the quantitative functional lymphoscintigraphy is really giving objective and reproductive parameters to evaluate a treatment efficacy (decongestive physiotherapy, surgery, drugs) in lymphedemas useful to assess new lymphotonic treatments. PMID- 10378341 TI - [Current status of therapy of lymphedema in Austrian hospitals--a comprehensive Austrian survey]. AB - The overall Austrian survey of the Austrian Lymph-Liga on the acutal state of the diagnosis and therapy of the hospitals of Austria is represented by making enquiries in 178 hospitals. The response was moderate at 34%, with only 46 hospitals offering a therapy of lymphedemas (approximately 26%). The type of therapy does not seem standardized at the Austrian hospitals. This explains a high number of multiple entries. The conservative therapy recognized internationally today, or rather, the complex-2-stage-decongestive physiotherapy (CDP), is only mentioned in 24% (including multiple entries). The time required per patient per day, the cyclical weekly reception and the total time of the decongestive therapy are very varied compared to the standards recommended today. Personal qualification of the lymph-therapists is given by physiotherapists in the majority. The specialist competence is very different, but it is identified as competent in the majority through specialists of the internal and of physical medicine. Most of the cost of medical care is covered by Compulsory Insurance. The medical care is limited in every second case (it could not be clarified whether there are organisational reasons, staff-related reasons, or financial reasons). Supplementary data of a "Quality of life" survey among lymphedema patients underline the necessity of a concept for standards, diagnosis and therapy. More than half of the patients asked (65%) complained about a time period of 5 to 10 years from the beginning of the illness till the beginning of a definitive therapy. Every other patient does not feel optimally treated and is under an increased pressure of suffering. PMID- 10378342 TI - Health related behaviors and cancer screening of lesbians: results from the Boston Lesbian Health Project. AB - The purpose of this article is to present data on lesbian health-related and cancer screening behavior. This is an area in which not a great deal of data exist and which is particularly interesting in view of previous data suggesting that lesbians do not seek routine services because of a fear of homophobia. This paper discusses a portion of a larger survey completed by a national community based lesbian sample. The results show that the lesbians in this sample have healthy behaviors in general and utilize routine health screening. There is some indication that alcohol use is heavier in this sample than among women in general, an area that warrants further investigation. PMID- 10378343 TI - Managed care organizations and mammography: opportunities to serve underserved women. AB - Breast cancer is one of the major causes of mortality and morbidity among women. Breast cancer screening (mammography) has been shown to be an effective preventive service. Significant proportions of women for whom mammography would be an appropriate intervention, especially older, low-income, and minority women, do not receive it. A large proportion of American women (including those in the workforce or who are Medicare and Medicaid beneficiaries) is now enrolled in managed care plans and that trend is likely to continue. Analysts have identified several concerns related to access and use of preventive services by low-income and other vulnerable populations. Research related to these concerns is summarized. Many research-based interventions have been identified that increase the likelihood of women receiving mammography. These are summarized and recommendations are made for managed care organizations to implement them. PMID- 10378344 TI - Breast screening practices among Russian immigrant women in Israel. AB - This article discusses cognitions, attitudes, and practices aimed at secondary prevention of breast cancer in the national sample of 620 women aged over 35 who immigrated to Israel from the former Soviet Union after 1989. The study has shown that universal access to preventive care may not translate into its optimal utilization among the marginalized population groups. Specifically, while being at moderate to high cancer risk, Russian immigrants refrain from screening activities--gynecological check-ups, self- and clinical breast exams and mammography. This is a drastic reversal of the pre-emigration pattern: two-thirds of respondents underwent breast screening in the past and only one-third sustained this practice in Israel. The risk groups for late detection of breast cancer are the women least integrated into the host society: those over age 60, unemployed or having unskilled jobs. Women who had no regular primary care providers showed the lowest cancer awareness and minimal screening activity. Even those who knew key breast cancer facts, considered themselves susceptible, and realized the role of early detection in practice did little to avert the danger. Three explanations of the gap between the cognitions and practice are suggested: (a) immigrants' low health motivation, reflecting their downward social mobility and preoccupation with resettlement problems; (b) low self-efficacy and external locus of control over health typical for ex-Soviet citizens; and (c) communicative and other cultural barriers to health care services. PMID- 10378345 TI - Health behavior change models and their socio-cultural relevance for breast cancer screening in African American women. AB - Models of health behavior provide the conceptual bases for most of the breast cancer screening intervention studies. These models were not designed for and have not been adequately tested with African American women. The models discussed in this paper are: The Health Belief Model, the Theory of Reasoned Action/Theory of Planned Behavior, and the Transtheoretical Model. This paper will examine the socio-cultural relevance of these health behavior models, and discuss specific socio-cultural dimensions that are not accounted for by these paradigms. It is critical that researchers include socio-cultural dimensions, such as interconnectedness, health socialization, ecological factors and health care system factors into their intervention models with African American women. Comprehensive and socio-culturally based investigations are necessary to guide the scientific and policy challenge for reducing breast cancer mortality in African American women. PMID- 10378346 TI - Psychological issues in genetic testing for breast cancer. AB - Genetic testing for inherited forms of breast cancer is currently available to individuals who want to learn their genetic status for the BRCA1 and BRCA2 genes. Although still largely limited to research programs, widespread commercial testing and incorporation of genetic testing into primary care practices will occur in the not too distant future. Despite the availability of this technology, treatment and prevention strategies offered to these often healthy women are limited and somewhat controversial. Due to the medical and emotional complexities associated with the gap between genetic information and treatment interventions, behavioral scientists are currently investigating the psychosocial implications involved in genetic testing for the BRCA1/2 genes. This article attempts to summarize the current research models, and reviews the most recent findings of investigations evaluating the emotional and behavioral implications associated with genetic testing for breast cancer susceptibility. PMID- 10378347 TI - Screening for breast and cervical cancers: the importance of knowledge and perceived cancer survivability. AB - INTRODUCTION: This study examines the association between recent screening for breast and cervical cancers, knowledge of cancer risk factors, and perceptions of surviving cancer. METHODS: Data were from the Cancer Control Supplement to the 1992 National Health Interview Survey (NHIS-CCS). The dependent variable combined breast and cervical cancer screening practices into a single composite index. Two independent variables combined women's knowledge about breast and cervical cancers into single indicators--one representing risk factor knowledge, the other representing perceived likelihood of surviving breast and cervical cancers following early detection. RESULTS: Multivariate analysis showed that recency of screening for both breast and cervical cancers was associated with knowledge of cancer risk factors and perceptions of surviving cancer. Education, household income, and smoking status also were correlates of comprehensive screening. Significant interactions between income and perceived survivability, and between education and perceived survivability suggested that the effects of income and education on comprehensive screening varied with perceptions about surviving cancer. CONCLUSION: The study suggests that knowledge and attitudinal questions can be combined for two diseases to enhance understanding of who is most likely to be screened comprehensively for breast and cervical cancers. Although national trends show that large percentages of women over age 50 are having mammograms and Pap tests, this progress is not likely to be sustained unless existing barriers are eliminated. Limited knowledge about breast and cervical cancer risk factors and misperceptions about survival from cancer represent two of these barriers. PMID- 10378348 TI - Breast cancer in mass circulating magazines in the U.S.A. and Canada, 1974-1995. AB - This paper presents the results of a study of the images of breast cancer in the highest circulating periodicals in the USA and Canada over a twenty year period of time. Both manifest and latent themes are noted and described. The emphasis in the manifest themes is on the medical aspects of the treatment and early detection of breast cancer. The latent themes emphasize the contrast in the ways that women with the disease, as compared to their doctors, are described. Notably, women are portrayed as being 'worried about their health' and, in particular, the most feared of 'their' diseases, breast cancer. Breast cancer is said to be caused by everything, especially women's own traitorous bodies. Women are described as isolates, as emotional and preoccupied with their sexual attractiveness. Doctors are described in contrasting ways, as moral truth seekers, infused with rationality and intelligence. The ubiquitous causes of breast cancer are also noted. The paper concludes with a discussion of the possible implications of the gendered character of the reporting about breast cancer. PMID- 10378349 TI - Pathobiology of Helicobacter pylori infection. PMID- 10378350 TI - Helicobacter pylori and apoptosis. AB - In an attempt to understand the diverse effects of infection with Helicobacter pylori on epithelial mucosal mass and consequent clinical outcome, the relationship between H. pylori infection and gastric epithelial cellular turnover has been investigated. Our results indicate that H. pylori increases epithelial cell proliferation and apoptosis in vivo, but that infection with bacteria of the cagA genotype leads to relatively more proliferation than apoptosis. This review explores the causes of the induction of apoptosis in gastric epithelial cells by H. pylori and the consequences of alterations in apoptosis to the maintenance of gastric mucosal homeostasis. PMID- 10378352 TI - Accuracy and economics of Helicobacter pylori diagnosis. AB - Many diagnostic tests are available to establish Helicobacter pylori infection status. Most of the tests are accurate though none works perfectly, and no gold standard for diagnosis exists. Newly developed serum immunoassay kits can substitute for laboratory-based enzyme-linked immunosorbent assays, but whole blood immunoassays do not yet demonstrate adequate performance characteristics. Serologic diagnosis of H. pylori remains the most cost-effective option and should be utilized to establish initial infection in the majority of cases. If rapid urease testing is performed at endoscopy, negative results can be confirmed with a subsequent serologic test in those patients with a high probability of infection. Obtaining additional gastric tissue at endoscopy to evaluate for bacterial infection is reasonable if specimens are being taken for a mucosal defect. Confirmation of bacterial eradication cannot be justified for all post treatment patients at present due to the expense. It is important to test for cure in those patients with complicated ulcer disease and those with recurrent symptoms after therapy. PMID- 10378351 TI - Structure, function and localization of Helicobacter pylori urease. AB - Helicobacter pylori is the causative agent of most cases of gastritis. Once acquired, H. pylori establishes chronic persistent infection; it is this long term infection that, is a subset of patients, leads to gastric or duodenal ulcer, gastric cancer or gastric MALT lymphoma. All fresh isolates of H. pylori express significant urease activity, which is essential to survival and pathogenesis of the bacterium. A significant fraction of urease is associated with the surface of H. pylori both in vivo and in vitro. Surface-associated urease is essential for H. pylori to resist exposure to acid in the presence of urea. The mechanism whereby urease becomes associated with the surface of H. pylori is unique. This process, which we term "altruistic autolysis," involves release of urease (and other cytoplasmic proteins) by genetically programmed autolysis with subsequent adsorption of the released urease onto the surface of neighboring intact bacteria. To our knowledge, this is the first evidence of essential communal behavior in pathogenic bacteria; such behavior is crucial to understanding the pathogenesis of H. pylori. PMID- 10378353 TI - Realities of diagnosing Helicobacter pylori infection in clinical practice: a case for non-invasive indirect methodologies. AB - BACKGROUND: The current, arbitrarily defined gold standard for the diagnosis of H. pylori infection requires histologic examination of two specially stained antral biopsy specimens. However, routine histology is potentially limited in general clinical practice by both sampling and observer error. The current study was designed to examine the diagnostic performance of invasive and non-invasive H. pylori detection methods that would likely be available in general clinical practice. METHODS: The diagnostic performance of rotating clinical pathology faculty using thiazine staining was compared with that of an expert gastrointestinal pathologist in 38 patients. In situ hybridization stains of adjacent biopsy cuts were also examined by the expert pathologist for further comparison. Receiver operator characteristic (ROC) analysis was performed to evaluate whether the diagnostic performance of the expert pathologist differed depending upon the histologic method employed. A similar analysis was made to evaluate the diagnostic performance of pathology trainees relative to the expert. In the absence of an established invasive gold standard, non-invasive testing methods (rapid serum antibodies, formal Elisa antibodies and carbon-14 urea breath testing) were evaluated in 74 patients by comparison with a gold standard defined using a combination of diagnostic tests. RESULTS: Using either rapid urease testing of biopsy specimens or urea breath testing as the gold standard for comparison, the diagnostic performance of the rotating clinical pathology faculty was inferior to that of the expert gastrointestinal pathologist especially with regard to specificity (e.g., 69 percent for the former versus 88 percent, with the latter relative to rapid urease testing). Although interpretation of in situ hybridization staining by the expert appeared to have an even higher specificity, ROC analysis failed to show a difference. The mean ROC areas for thiazine and in situ hybridization staining for trainee pathologists relative to the expert were 0.88 and 0.94, respectively. In untreated patients, urea breath testing had a sensitivity and specificity of 100 percent as compared with thiazine staining with a sensitivity of 83 percent and a specificity of 97 percent. Post-therapy, breath testing had a sensitivity of 100 percent but a specificity of only 86 percent as compared with invasive testing with a sensitivity and specificity of 100 percent. Rapid serum antibody testing and formal Elisa antibody testing agreed in 93 percent of cases (Kappa 0.78) with the rapid test being correct in three of the four disagreements. CONCLUSIONS: The current study illustrates a number of realities regarding H. pylori diagnosis. There is no diagnostic gold standard in general clinical practice. Accurate interpretation of specially stained slides is a learned activity with a tendency towards overdiagnosis early on. Urea breath testing is likely to be the diagnostic method of choice for untreated patients in general clinical practice although antibody testing is almost as accurate. Rapid antibody tests are at least as accurate as formal Elisa antibody tests. Urea breath testing is useful for confirming cure after therapy, but false-positive results may occur in some patients. PMID- 10378354 TI - Helicobacter pylori and smoking: two additive risk factors for organic dyspepsia. AB - The hopes to distinguish between organic and functional dyspepsia on the grounds of the patient's symptomatology have not been fulfilled due to the low specificity of the so-called sinister symptoms. There is increasing evidence accumulating that Helicobacter pylori status and other environmental factors such as smoking have a higher discriminant power. Studies performed in our laboratories testing H. pylori status on gastric biopsy samples have shown that preselection of patients according to smoking habits and H. pylori status has a higher potential in avoiding unnecessary endoscopies in primary care patients as compared to risk factors based on patient complaints. Out of a total population of 282 primary care patients, one out of 24 endoscopies revealed significant pathology such as peptic ulcer or reflux esophagitis in the non-smokers with a negative H. pylori status, but when both risk factors were positive, the percentage rose to one out of every two patients. These observation have largely been confirmed by recent studies where H. pylori status was prospectively assessed prior to endoscopy by highly specific H. pylori serology or 13C breath test analysis. PMID- 10378355 TI - Helicobacter pylori and non-steroidal anti-inflammatory drugs: does infection affect the outcome of NSAID therapy? AB - 1. H. pylori gastritis appears to increase the likelihood of developing dyspeptic symptoms on NSAID therapy. 2. There is preliminary evidence that the histologic severity of H. pylori gastritis may be adversely affected by NSAID therapy, with a consequent increase in the risk of developing a peptic ulcer, possibly with complications. Whether this results from an effect on the inflammatory process or results from a quantitative increase in H. pylori colonization is unknown. In these respects, ASA may differ from other NSAIDs. 3. Ulcers are more likely to develop during the course of NSAID therapy in those infected with H. pylori; eradication of the infection reduces ulcer recurrence in the face of continued NSAID therapy, and it seems likely that this must reduce but not abolish the risk of GI bleeding in those using NSAIDs. Eradication also reduces the damage (and possibly risks) of low-dose aspirin therapy. 4. While H. pylori and NSAID use are independent risk factors for GI bleeding, whether or not they are interactive remains unresolved. 5. The effect of H. pylori infection on the risk of perforation during NSAID therapy, or conversely, the contribution of NSAID therapy to the risk of perforation in H. pylori-infected subjects, is also unclear at the present time. 6. Only large outcome studies of accurately diagnosed patients (with regard to H. pylori gastritis), and with much more specific detail as to the type of NSAID, dose and duration of therapy, employing only well-defined end-points, such as significant hemorrhage, perforation or death, and avoiding all surrogate markers short of these end points can hope to unravel this tangled web. PMID- 10378356 TI - Helicobacter pylori: therapeutic targets. AB - Helicobacter pylori is now considered a major pathogen of the upper gastrointestinal tract. It is seen as an important cause of peptic ulceration not associated with NSAID use. It is also increasingly linked to other diseases of the GI tract, although the relationship between the organism and conditions such as gastric cancer, non-ulcer dyspepsia and gastroesophageal reflux disease is not as clear as is the case in peptic ulcer disease. This is probably because of a lack of well-performed, statistically powerful, prospective therapeutic trials that indicate that H. pylori eradication is of benefit in these diseases. The high infection rate without overt disease seen in many populations, especially from developing countries, probably contributes to this "credibility gap." While we have excellent therapeutic regimens available at this time, rational targeting requires that the objective evidence in favor of therapeutic intervention in upper GI disease, as well as the local H. pylori epidemiology, needs to be considered. PMID- 10378357 TI - Treatment of H. pylori infection: the reality. AB - Despite the wide dissemination of information on Helicobacter pylori, there is still a great deal of variation in how general practitioners treat the infection and in which circumstances they prescribe eradication therapy for H. pylori. Specialty societies have developed consensus guidelines that recommend a strategy to test and treat dyspeptic patients for H. pylori infection although the data to support these recommendations are weak at the present time. As a result, there is still confusion about the indications for treatment and the treatment regimens that are likely to be effective in routine clinical practice. PMID- 10378358 TI - Eradication of Helicobacter pylori infection in the management of patients with dyspepsia and non-ulcer dyspepsia. AB - Although H. pylori infection has been recognized as a major etiological agent for the development of chronic active gastritis, duodenal ulcer and benign non-NSAID related gastric ulcer, its role in the development of symptoms in patients with dyspepsia remains uncertain. Results from population-based epidemiological studies have been conflicting regarding a causal link between H. pylori infection and dyspepsia. Abnormalities in gastric acid secretion may exist in some dyspeptic patients. Whether disordered gastric motility seen in dyspeptic patients is related to the infection is not clear based on the results in the literature. Numerous clinical trials have been undertaken to eradicate H. pylori infection and improve the symptoms in dyspeptic patients; however, the results have been discrepant between studies. Many published studies suffer from methodological problems that have made interpretation difficult. Large, well conducted, randomized, placebo-controlled, clinical trials with long-term follow up are needed to justify the beneficial effect of H. pylori eradication treatment in dyspeptic patients seen in some small studies. H. pylori eradication therapy is cost-effective in H. pylori-infected dyspeptic patients although this benefit may take a long time to accrue, especially in younger patients. PMID- 10378359 TI - Helicobacter pylori: the Middle East scenario. AB - A review of Helicobacter pylori in the Middle East is presented. Prevalence studies have been performed in asymptomatic population groups from Algeria, Israel, Saudi Arabia and Turkey. These showed that the prevalence of H. pylori is similar to that of the developing countries of the world with a high level of infection in childhood (40 to 70 percent), which increases with age to 85 to 90 percent. Israel, however, has a low prevalence in children (10 percent), but there is a rapid rise in the second decade of life to 39 percent, reaching 79 percent in those over 60 years old. The prevalence rates were higher in those living in communal settlements (72 percent) than in urban dwellers (65 percent). The infection rates were higher in persons of Mediterranean and Asian origin (89 percent) compared to those of Western European/North American origin (57 percent). The prevalence rate of H. pylori infection in patients undergoing endoscopy for upper gastrointestinal symptoms has now been reported from many Middle Eastern countries, including Egypt, Iran, Israel, Oman, Saudi Arabia, the United Arab Emirates and Yemen. These studies showed that patients with gastritis and peptic ulcer disease had similar rates of infection as reported from Europe, United States and Africa (71 to 92 percent). However, patients with non-ulcer dyspepsia had higher rates of infection (61 to 89 percent). The H. pylori scenario from the prevalence rates, treatment protocols and responses to treatment does not differ very much from other developing areas of the world. PMID- 10378360 TI - Does Helicobacter pylori infection contribute to gastroesophageal reflux disease? AB - Helicobacter pylori organisms that infect the stomach conceivably could contribute to esophageal inflammation in patients with gastroesophageal reflux disease (GERD) through any of at least three potential mechanisms: 1) by causing an increase in gastric acid secretion; 2) by spreading to infect the gastric-type columnar epithelium that occasionally can line the distal esophagus; and/or 3) by secreting noxious bacterial products into the gastric juice. Studies regarding these potential mechanisms are discussed in this report. Most investigations have found no apparent association between H. pylori infection and reflux esophagitis. Presently, infection with H. pylori does not appear to play an important role in the pathogenesis of GERD. PMID- 10378361 TI - [Coping with stress and personality structure in patients with and without stress associated symptoms of gastroesophageal reflux]. AB - BACKGROUND: Psychological aspects like stress, emotions or personality are known to affect the severity of symptoms of gastroesophageal reflux disease (GERD). The aim of the present study was to evaluate differences in coping with stress, structure of personality and also objective and subjective parameters of patients with or without a stress-related perception of symptoms in GERD. METHODS: 100 patients which underwent laparoscopic antireflux surgery at our department of surgery were included in this study. All patients answered questionnaires to evaluate their coping with stress (SVF), structure of personality (FPI-R) and quality of life (GILQI) pre- and postoperatively. Also data of physiological parameters like manometry, 24-hours pH monitoring, endoscopy and clinical history were included. Patients were divided into two groups: one with and one without a stress-related perception of symptoms. RESULTS: 46 out of 100 patients declared stress-related symptoms of reflux (group 1) and 54 out of 100 had no influence of stress to their reflux disease (group two). Those two groups showed significant differences (p < .05) in some coping strategies and their personality: Coping with stress (SVF): Trial to control the situations (18.3 vs. 13.1), trial to control reactions (18.2 vs. 13.3), requirement of social support (9.7 vs. 14.8), tendency to escape (7.7 vs. 13.9) and aggression (13.1 vs. 7.6); structure of personality (FPI-R): Standard of performance (10.8 vs. 7.2), stress (9.8 vs. 4.7) and physical discomfort (7.6 vs. 4.6). We also found significant (p < .05) differences in pre- and postoperative quality of life (GILQI: preoperative 86.3 vs. 98.5 points; postoperative 117.9 vs. 128.2 points) and the day-time of reflux perception. There were no differences in physiological parameters. CONCLUSION: These findings point out that there are no physiological differences between the two groups with or without stress-related symptoms in GERD. But we found significant differences in psychological factors. Therefore we suggest that preoperative psychological interventions may optimize the subjective outcome after antireflux surgery in patients with a stress-related perception of symptoms. PMID- 10378362 TI - [Effect of stomach motility on measuring stomach permeability with saccharose in vivo]. AB - Determination of the urinary excretion of sucrose after an oral dose has been used as a noninvasive test to measure gastric permeability in several clinical studies. Regarding different contact times of sucrose solution within the gastric mucosa, the present study investigates a possible influence of the gastric emptying rate on the sucrose permeability test. Urinary sucrose excretion and the gastric emptying rate of liquids using 13C-acetate breath test were determined in twelve healthy volunteers. Furthermore, in seven volunteers gastric emptying was accelerated by intravenous erythromycin and prolongated by oral anticholinergic propantheline in nine healthy controls. Breath samples were measured using infrared spectroscopy. The half-emptying time and Lag-phase were correlated with the urinary sucrose excretion. Erythromycin caused a significant (p = 0.02) reduction of the half-emptying time (median 35.0 min) compared with untreated controls (median 59.9 min), whereas propantheline significantly increased the half-emptying time (median 69.4 min, p = 0.01). After pharmacological increase of the half-emptying time the urinary sucrose excretion only slightly differs from the sucrose excretion of controls (median [range] 0.057 [0.034-0.106]% versus 0.031 [0.017-0.162]%), but there was an increase of urinary sucrose excretion in probands following reduction of the half-emptying time with erythromycin (0.077 [0.023-0.221]%. The present study shows that gastric motility has a possible influence on the sucrose permeability test. The sucrose permeability has to be interpreted critically concerning its clinical use especially in patients with altered gastric motility. PMID- 10378363 TI - Budd-Chiari syndrome in a patient with factor V Leiden--successful treatment by TIPSS placement followed by liver transplantation. AB - The causes of Budd-Chiari syndrome (BCS) comprise several diseases leading to thrombophilia. One of the most common thrombophilic disorders is resistance against activated protein C, caused by a single point mutation of the factor V gene. In December 1993, a 22-year-old patient was given a diagnosis of subacute BCS with occlusion of all major hepatic veins. Placement of a transjugular intrahepatic portosystemic stent shunt led to rapid disappearance of ascites and hepatic encephalopathy. During the following two years, recurrent partial occlusions of the shunt were treated by balloon angioplasty. The cause of the BCS still being unknown, in October 1996 we performed extensive laboratory investigations concerning states of thrombophilia and found moderately elevated IgG anticardiolipin antibodies (19.7 U/ml) and a resistance against activated protein C caused by heterozygosity for a point mutation of the factor V gene (1691G-->A; factor V Leiden). As a consequence, oral anticoagulation with coumarin was initiated. In October 1997, elective liver transplantation was performed which led to disappearance of APC resistance. Moreover, IgG anticardiolipin antibodies have been negative since then. If BCS is caused by APC resistance, liver transplantation not only treats the chronic liver disease but also cures the state of thrombophilia since factor V is mainly synthesized in the liver. PMID- 10378364 TI - [46-year-old patient with polyneuropathy, hepatosplenomegaly, endocrinopathy, M gradient, skin manifestations, sclerotic bone changes and therapy refractory ascites]. AB - The POEMS syndrome is a rare multisystemic disorder with polyneuropathy, organomegaly, endocrinopathy of various forms, production of monoclonal (M) component, and skin changes. We describe a 46-year-old man who developed ascites one year after the onset of peripheral neuropathy with accompanying muscle atrophies and increasing weakness. Extensive evaluation revealed that the patient had no underlying liver disease, malignancy, infection, or cardiac or renal disease. The ascites initially responded to high-dose corticosteroid therapy. The patient had many clinical features of the described POEMS syndrome including sclerotic bone lesions, a persistent lambda-paraprotein and refractory ascites. In this case ascites was a main presenting feature. Thus, the POEMS syndrome must be added to the list of rare causes of refractory ascites. PMID- 10378365 TI - [Pouch stomach reconstruction after gastrectomy]. AB - Up to now there is no general agreement on the ideal reconstruction after total gastrectomy. The importance of the duodenal passage, the need for a pouch reconstruction, and the ideal pouch volume are matters of controversy. Prospective randomized trials show a significantly better quality of life, a higher body weight and a better glucose regulation in patients with a curative operation and good life expectancy, if the duodenal passage is preserved. Reconstruction with a small jejunal pouch offers a better reservoir, less reflux and a better nutritional passage, but a statistically significant improvement of life quality could not be demonstrated up to now. Nevertheless, patients with a curative resection should undergo pouch reconstruction with preservation of the duodenal passage. If curative resection is not possible, reconstruction can be performed according to Hunt-Lawrence-Rodino. The Roux-en-Y-reconstruction without pouch should only be performed in high-risk patients and in carcinoma of the cardia with intrathoracic anastomosis. Nevertheless, further prospective randomized studies with more patients and more specific tests to measure life quality are necessary to evaluate the importance of a jejunal pouch in patients with a preserved duodenal passage. PMID- 10378366 TI - [Therapy of Wilson disease]. AB - Wilson disease is a copper storage disease with autosomal-recessive trait that is predominantly a disorder of the adolescent and young adult. Clinical manifestations are dominated by hepatic and/or neurological symptoms. Diagnostic procedures include determination of total serum copper, free serum copper and serum ceruloplamin concentrations as well as urinary copper excretion. Confirmation of diagnosis may be achieved by liver biopsy and histological determination of copper content. The aim of treatment is reduction of tissue copper concentration and detoxification of copper. Drugs applied are the chelating agents. D-penicillamine and trientine, or zinc. The chelating agents induce renal and biliary copper excretion and increased synthesis of metallothionein, which attaches and detoxifies intracellular copper, leading to impaired absorption and binding of excess intracellular copper. Treatment with zinc results in induction of hepatic and intestinal metallothionein synthesis. Regular examinations of the parameters of copper metabolism are necessary in order to control the therapeutic effect. Free copper serum concentrations and urinary copper excretion should reach values below 10 micrograms/dl and 80 micrograms/day, respectively. A significant improvement of clinical symptoms and normalization of parameters of copper metabolism can be expected earliest six months after onset of therapy. Anti-copper treatment may be accompanied by copper reduced diet. Lifelong therapy is required and provides life-expectancy near to normal. Interruption of treatment leads to reaccumulation of copper, often resulting in fulminant hepatic failure. This can also be observed as initial presentation in 5% of cases (predominant age 12 to 25 years). End stage liver disease and fulminant hepatic failure are indications for liver transplantation by which the genetic defect is phenotypically cured. Here decoppering treatment is no longer required. Whether severe neurological disorders may also be improved is not clear until today. PMID- 10378367 TI - [Guidelines of GASL for nutrition in liver diseases and liver transplantation]. PMID- 10378368 TI - [ICAM-1 antisense therapy: a new treatment concept on Crohn disease? Placebo controlled study of ICAM-1 antisense oligonucleotide therapy in Crohn disease]. PMID- 10378369 TI - [Incentive for rethinking--early enteral nutrition in patients with pancreatitis]. PMID- 10378370 TI - [Pathogenetic germ-line mutation in patients with hereditary nonpolyposis colorectal carcinoma (HNPCC)]. PMID- 10378371 TI - Apoptosis of T lymphocytes in acute disseminated encephalomyelitis. AB - Apoptosis has been shown to be an efficient mechanism involved in clearance of T lymphocytes from the brains of animals with acute experimental autoimmune encephalomyelitis (EAE), an animal model for human multiple sclerosis. In this report we describe a case of acute disseminated encephalomyelitis following general measles infection. In this disease, which closely mimics the pathology of acute EAE we found a high percentage (30%) of apoptotic T cells. This indicates that in both rodent and human brain clearance of T cell-mediated inflammation follows similar mechanisms. PMID- 10378372 TI - Quantitative analysis of NF1 and OMGP gene transcripts in sporadic gliomas, sporadic meningiomas and neurofibromatosis type 1-associated plexiform neurofibromas. AB - The close association of neurofibromatosis type 1 (NF1) with gliomas raises the question of whether the NF1 gene may be involved in the pathogenesis of sporadic astrocytic brain tumors. However, no frequent mutations within NF1 have been described in these tumors. Recent data on a limited series of gliomas indicate that NF1 expression may even be increased, thereby questioning the role of NF1 as a tumor suppressor in astrocytomas. In the present study, we examined the expression of NF1 in a series of 96 tumors including astrocytomas, meningiomas and plexiform neurofibromas. NF1 RNA transcription levels were compared to those of the reference genes B2M, ACTB and GAPD. The expression of OMGP, which is interposed in the NF1 gene, served as an additional control. NF1 expression did not significantly diverge among different malignancy stages of astrocytomas. As expected, the plexiform neurofibromas showed only very low NF1 expression. A striking finding was the highly variable expression of those genes selected to serve as references. While B2M and ACTB exhibited comparable levels of expression within different grades of astrocytomas and meningiomas, GAPD showed an inverse pattern in these tumors. In conclusion, NF1 expression is strongly reduced in NF1 associated plexiform neurofibromas but not in astrocytic tumors. The significant differences between B2M, ACTB and GAPD transcript levels brings into question the common practice of defining gene expression as a ratio between the transcripts of interest and those of these reference genes. PMID- 10378373 TI - Distal axonopathy does not occur without neurofilament accumulation in gamma diketone neuropathy: comparative studies of normal and neurofilament-deficient quail. AB - The neurotoxic effects caused by chronic exposure to 2,5-hexanedione (2,5-HD) were investigated in normal and neurofilament (NF)-deficient quail (Quv strain). These quail were given 175 mg/kg per day of 2,5-HD intraperitoneally for 24 weeks. Five of nine normal quail showed clumsy gait. They had NF-rich axonal swellings in the distal parts of the peripheral nerves, ventral and lateral funiculus of the cervical cord, and cerebellar peduncles. Axonal degeneration consisting of accumulation of mitochondria, vesicles, microtubules and dense bodies was found distal to the axonal swellings. Testicular atrophy appeared in two normal quail. In contrast, four of nine Quv quail showed systemic tonic convulsion, and died of respiratory paralysis within 6 days. No significant changes in the nervous system or testis of these four dead Quv quail. The five other Quv quail survived and did not show any neurological signs. Again, no significant changes were detected in the nervous system or testis of the surviving Quv quail. The present study revealed that distal axonal degeneration did not occur without NF accumulation. These results suggest that NF accumulation is an essential factor in the development of distal axonopathy in gamma-diketone neuropathy. PMID- 10378374 TI - Focal pathology in the Edinger-Westphal nucleus explains pupillary hypersensitivity in Alzheimer's disease. AB - Patients who suffer from Alzheimer's disease (AD) and a sub-population of community-dwelling elders show an exaggerated pupillary reaction to dilute tropicamide, a cholinergic antagonist. This finding may serve as an early diagnostic marker of AD. Here we report a likely pathological basis for this hypersensitive pupillary response. Our observations indicate that the Edinger Westphal nucleus (EW), a known center for the control of pupillary function, is a selective target of Alzheimer pathology early in the course of the disease. In all AD cases examined, the EW contained plaques and tangles. In contrast, the adjacent somatic portion of the oculomotor complex was virtually spared of pathology. Early pathology in the EW is likely to initiate a cascade of events that may give rise to pupillary hypersensitivity. PMID- 10378375 TI - Immunohistochemical and ultrastructural characterization of neuritic clusters around ghost tangles in the hippocampal formation in progressive supranuclear palsy brains. AB - We performed a detailed study of swollen neurite aggregation surrounding extracellular neurofibrillary tangles (ghost tangles, GTs) in brains of patients with progressive supranuclear palsy (PSP) by immunohistochemistry and electron microscopy (EM). The complex structures, designated as tangle-associated neuritic clusters (TANCs), were found in the hippocampus and parahippocampal cortex in all five PSP brains examined. TANCs measured from 20 to 40 microm across; twice as large as nearby neurons. Each neurite was globular or fusiform in shape, measured up to 10 microm in diameter, and was found between loosened fascicles of GTs or along their outer rims. There were several subsets of neurites that were argyrophilic or immunoreactive against antibodies to either phosphorylated tau protein, phosphorylated neurofilaments, ubiquitin, or synaptophysin. On EM, TANCs consisted of numerous axon terminals of varying size, which were filled with flocculate dense bodies, vesicular profiles, and synaptic vesicles, as well as normal-looking and degenerating cell organelles. Some axons had 13- to 15-nm thick straight tubules that showed tau immunoreactivity; however, there was little neurofilament accumulation. Most of the swollen axon terminals conformed to the ultrastructural features of either reactive or degenerating terminals. The neurites identified by immunohistochemistry only represented a minority of the swollen axons visualized by EM. Tubules of GTs were dispersed in the extracellular space, but no amyloid fibrils were found. TANCs may constitute a distinctive form of neuronal degeneration in PSP cortices. We hypothesize that axon terminal accumulation may occur in response to GT-formation. PMID- 10378376 TI - Preferential neurodegeneration in the cervical spinal cord of progressive supranuclear palsy. AB - Spinal cord lesions have seldom been described in cases with progressive supranuclear palsy (PSP). We thus decided to analyze spinal cord lesions by microtubule-associated protein 2 (MAP2) immunohistochemistry in six cases of PSP, five cases of Parkinson's disease (PD) and two cases of corticobasal degeneration (CBD), all of which cause parkinsonism, while six patients without any neurological disease served as controls. In the PSP cases, the MAP2 expression in the cervical spinal cords significantly decreased in the medial division of the anterior gray horn, intermediate gray and posterior gray horn, but showed no significant change in the substantia gelatinosa and lateral division of the anterior gray horn. The thoracic and lumbar spinal cords were well preserved for MAP2 immunoreactivity. In addition, the globose type neurofibrillary tangles and glial fibrillary tangles were more conspicuous in the cervical than in the thoracic and lumbar spinal cord in PSP cases. On the other hand, the PD and CBD cases showed no significant decrease of MAP2 immunoreactivity in the spinal cords. The small neurons, which are located rather selectively in the intermediate zone of the spinal cord, are considered to be mostly present in the interneurons, and are also thought to play a role in various types of focal dystonia, such as neck dystonia. We therefore consider the distinct decrease in the MAP2-positive neuronal processes in the cervical spinal cord may partly reflect the loss of interneurons and may, thereby, possibly cause nuchal dystonia. PMID- 10378377 TI - Microtubule-associated protein tau, heparan sulphate and alpha-synuclein in several neurodegenerative diseases with dementia. AB - Microtubule-associated protein tau forms neurofibrillary lesions in Alzheimer's disease and several other neurodegenerative disorders, such as Niemann-Pick disease type C, subacute sclerosing panencephalitis, argyrophilic grain disease, myotonic dystrophy and motor neuron disease with neurofibrillary tangles. In this study we have compared the characteristics of tau pathology in these diseases using immunohistochemistry and phosphorylation-dependent and phosphorylation independent anti-tau antibodies. The pattern of staining for heparan sulphate and alpha-synuclein was also investigated. We show that in all of these diseases tau deposits were stained by all anti-tau antibodies used, with the exception of argyrophilic grains which do not stain with antibody 12E8, confirming our previous findings. Heparan sulphate staining was present to a variable extent in all of these diseases, with the exception of subacute sclerosing panencephalitis, in which no staining was observed. Heparan sulphate staining coexisted with tau staining. In some cases it was more extensive than the tau staining. Alpha synuclein staining was present in presynaptic terminals with the exception of one case of Alzheimer's disease, in which alpha-synuclein-positive Lewy bodies were observed in the hippocampal formation. These findings indicate that tau deposits are antigenically similar in several neurodegenerative diseases and that tau staining is often associated with heparan sulphate staining, supporting the concept that heparan sulphate may be involved in the assembly of tau protein into filaments. PMID- 10378378 TI - Transgenic Lewis rats overexpressing the proteolipid protein gene: myelin degeneration and its effect on T cell-mediated experimental autoimmune encephalomyelitis. AB - Transgenic Lewis rats overexpressing proteolipid protein (PLP) genes in peripheral and central nervous myelin were produced by microinjecting murine genomic PLP sequences into fertilized eggs. The mouse PLP gene shares 98.7% homology in the nucleotide sequence with its rat counterpart, but both are fully identical on protein level. Homozygous rats show tremors early in postnatal life, eventually develop seizures, and die before they reach weaning age, while hemizygous animals are phenotypically normal and have a normal life expectancy. Transgene expression in the central nervous system (CNS) has profound consequences for myelin formation and maintenance: approximately twofold overexpression of PLP/DM-20, as seen in homozygotes, results in apoptosis of mature, and a developmental arrest of the remaining immature oligodendrocytes. Severe dysmyelination ensues, associated with reactive astrogliosis and microglia activation/proliferation. Activation of microglia is also prominent in hemizygous rats with low levels of transgene overexpression. In these animals, myelin sheaths remain intact, but there is low-grade myelin degeneration throughout life witnessed by myelin uptake and activation of microglia and astrocytes, in the absence of the expression of major histocompatibility complex class II gene products. There were no spontaneous lymphocytic infiltrates in areas of myelin degeneration. However, hemizygous LEW.PLP rats were more sensitive to experimental autoimmune encephalomyelitis mediated by T cells specific for PLP, but not another encephalitogenic myelin protein, MBP. PMID- 10378379 TI - Migratory potential of transplantable neural tumor cell lines. AB - Experimentally induced primitive neuroectodermal tumor (PNET) cell lines were transplanted into neonatal and adult rat brain and examined neuropathologically for their tumorigenic potential. Both cell lines showed a striking migratory behavior in both neonatal and adult brain. Migration of tumor cells was found in host brain parenchyma, along white matter tracts and associated with CSF pathways. These neural tumor cell lines provide a valuable tool for the development of strategies against strongly migrating neural tumors. PMID- 10378381 TI - Nerve cell loss in the thalamic mediodorsal nucleus in Huntington's disease. II. Optimization of a stereological estimation procedure. AB - This study provides the theoretical background of the decision to count approximately 750-1,300 neurons per individual in the preceding study of Heinsen et al. [6] finding a significant (P < 0.05) nerve cell loss in the thalamic mediodorsal nucleus in Huntington's disease with the so-called V(Ref) x N(V) method. Using a computer simulation of the study of Heinsen et al., it was shown that the legitimation for counting only 100-200 neurons per individual in previous studies comparable to that carried out by Heinsen et al. was based on incorrect assumptions. In this context it was of particular importance to confirm the theoretical prediction in the literature that the random error of total neuron number estimates obtained with the V(Ref) x N(V) method is actually greater than assumed in current stereological studies. In summary, this study revives the question of how many individuals need to be investigated and how many neurons (or other cell types, respectively) need to be counted per individual in studies comparable to that carried out by Heinsen et al. PMID- 10378380 TI - Nerve cell loss in the thalamic mediodorsal nucleus in Huntington's disease. AB - We estimated the total neurone number, glial number, and glial index (ratio glial cells/neurone) in the thalamic mediodorsal nucleus (MD) in seven patients suffering from Huntington's disease (HD; four males, three females, mean age 52.4 +/- 13.6 years) and age- and sex-matched controls (four males, three females, mean age 53.6 +/- 12.1 years) by means of a stereological protocol. The mean total neurone number (N(T)) in the MD of controls was 2,985,188 +/- 174,710, the mean glial number (G(T); astrocytes, oligodendrocytes) 21,785,008 +/- 2,986,678, and the glial index 7.29 +/- 0.88. In HD, the average neurone number was decreased by 23.8% to 2,275,321 +/- 247,162 (Mann-Whitney U-test P < 0.05), the mean glial number by 29.7% to 15,318,895 +/- 1,722,524 (Mann-Whitney U-test P < 0.05), the glial index was slightly reduced to 6.81 +/- 1.06. Gallyas' impregnation for the demonstration of fibrous astroglia gave strongly positive results in all cases with HD and negative results in the controls. The morpho functional correlation of the results is complicated because individual variability, presence of segregated and parallel neuronal circuits, and plasticity of the adult human CNS must be considered. PMID- 10378382 TI - Vascular changes in white matter lesions of Alzheimer's disease. AB - The pathogenesis of white matter lesions observed in Alzheimer's disease (AD) is not completely clear. We tested the hypothesis that white matter lesions are correlated with medullary artery sclerosis rather than with amyloid angiopathy. A total of 57 brains were examined, including 39 derived from patients with AD and 13 from patients with Binswanger's disease (BD) along with 5 from non neurological patients. Moderate or severe amyloid deposits in the meningocortical segment were observed in 32 out of 39 AD patients (82.1%), and in 2 out of 13 BD patients (15.4%). These deposits were not observed in the white matter segment, except for 2 patients with AD. The BD patients invariably had marked white matter lesions and fibrohyalinosis in the medullary arteries, with a mean sclerotic ratio of 48.1%. In contrast, the AD patients had mild or moderate white matter lesions and a sclerotic ratio of 37.9%, which was significantly greater than the controls. The scores for white matter lesions were correlated with the sclerotic ratio of the medullary arteries, but not with the ages of onset or the scores for amyloid angiopathy. Although amyloid angiopathy is an independent risk of white matter lesions, its role is limited in the pathogenesis of those associated with AD. Wall thickening of the medullary arteries, likely due to fibrohyalinosis, is closely correlated with the white matter lesions in AD, thus indicating a heterogeneity in its etiology. PMID- 10378383 TI - Glycosylation of microtubule-associated protein tau in Alzheimer's disease brain. AB - In the neurofibrillary pathology of Alzheimer's disease (AD), neurofibrillary tangles (NFTs) contain paired helical filaments (PHFs) as their major fibrous component. Abnormally hyperphosphorylated, microtubule-associated protein tau is the major protein subunit of PHFs. A recent in vitro study showed that PHF tangles from AD brains are highly glycosylated, whereas no glycan is detected in normal tau. Deglycosylation of PHF tangles converts them into bundles of straight filaments and restores their accessibility to microtubules. We showed that PHF tangles from AD brain tissue were associated with specific glycan molecules by double immunostaining with peroxidase and alkaline phosphatase labeling. Intracellular tangles and dystrophic neurites in a neuritic plaque with abnormally hyperphosphorylated tau, detected with the monoclonal antibodies AT-8 and anti-tau-2, were also positive with lectin Galanthus nivalis agglutinin (GNA) which recognizes both the N- and O-glycosidically linked saccharides. Colocalization was not seen in the extracellular tangles and amyloid deposition, suggesting that the glycosylation of tau might be associated with the early phase of insoluble NFT formation. Thus, although abnormal phosphorylation might promote aggregation of tau and inhibition of the assembly of microtubules, glycosylation mediated by a GNA-positive glycan appears to be responsible for the formation of the PHF structures in vivo. PMID- 10378384 TI - Local expression of monocyte chemoattractant protein-1 (MCP-1) in idiopathic inflammatory myopathies. AB - The idiopathic inflammatory myopathies (IIM), including dermatomyositis (DM), polymyositis (PM), and inclusion body myositis (IBM), are a group of autoimmune diseases characterized by the recruitment of lymphocytes and monocytes to the site of affection. The mechanism for recruitment of these cells likely involves chemokines. The monocyte chemoattractant protein-1 (MCP-1), a chemoattractant to T lymphocytes and monocytes, may play an important role in the pathogenesis of IIM. Frozen muscular tissues were obtained from eight cases of DM, five PM, and four IBM. We investigated the MCP-1 expression by the reverse transcription-PCR technique, immunohistochemistry and in situ hybridization. MCP-1 mRNA was markedly expressed in all the IIM cases. Greater amounts of MCP-1 mRNA were observed in DM, and in situ hybridization showed MCP-1 mRNA accumulation in perivascular mononuclear cells. Immunohistochemistry showed MCP-1 expression in vessels (endothelial cells and walls of veins and arteries) in all IIM cases. Very few mononuclear cells in DM perivascular infiltrates expressed MCP-1, whereas in PM and IBM, it was strongly expressed by mononuclear cells partially invading non-necrotic muscle fibers. These findings suggest that MCP-1 can contribute to the inflammatory response by attracting monocytes and T lymphocytes to sites of cell-mediated immune injury. The morphological characteristics and distribution of positive cells indicate that macrophages, lymphocytes, and endothelial cells previously reported to produce MCP-1, contribute to its production in IIM lesions. PMID- 10378385 TI - The heterogeneity in the immune response and efficiency of viral dissemination in brain infected with herpes simplex virus type 1 through peripheral or central route. AB - Using immunohistochemistry on adjacent brain sections, we studied the correlation between the dissemination of the virus, the inflammatory responses and the expression of major histocompatibility complex (MHC) proteins in rat brain infected with herpes simplex virus (HSV-1) F strain by either corneal scarification or intracerebral injection. Our results showed that the mortality of the corneally infected rats was much higher than that of the intracerebrally infected rats, due to a more extensive dissemination of the virus in the brain, particularly in the brain stem. The inflammatory responses were similar in brains infected through either route, as demonstrated by the expression of MHC I/II antigens on infiltrating lymphocytes, leukocytes and macrophage/microglia cells. While there was strong immunoreactivity for HSV-1 antigens in the cerebral cortex, the infiltrates were only located in subcortical areas, especially the hippocampus. Therefore, the distribution of these immune cells did not always overlap with the regions of viral infection. These results suggest that HSV-1 disseminate more efficiently from the peripheral to the central nervous system (CNS) than from CNS to CNS, which is independent of the immune responses, and that the cerebral cortex may immunologically respond to HSV-1 infection differently from other brain regions. PMID- 10378386 TI - Variable histological expression of dystrophinopathy in two females. AB - We report two carriers of Xp21 muscular dystrophy with unusual clinical manifestations and striking variability of dystrophin deficiency within the same muscle biopsy. The first patient was a 60-year-old nun with recent onset of cramps and proximal weakness, mimicking an acquired myopathy. Muscle biopsy disclosed slight alterations in one sample and severe dystrophic changes in another; dystrophin was absent in 7% fibers in the former specimen and in 60% in the second. X inactivation was skewed with 90% cells inactivating the same X chromosome. The second patient was a 17-year-old girl with hyperCKemia, learning disability and a family history of X-linked muscular dystrophy. Muscle biopsy displayed slight fiber size variability and some internal nuclei; dystrophin was absent only in one muscle fiber. A second sample with the same morphological features demonstrated dystrophin deficiency with mosaic distribution. The pattern of X inactivation was normal. These cases emphasize the variability of histopathological changes and dystrophin deficiency in Xp21 muscular dystrophy carriers and the risk of sampling errors in muscle biopsy. PMID- 10378387 TI - Berger disease: thirty years later. PMID- 10378388 TI - Congenital infection and B-cell responses in the newborn. PMID- 10378389 TI - Effects of a low-calorie diet on resting metabolic rate and serum tri iodothyronine levels in obese children. AB - The purpose of the study was to examine the effects of weight loss on resting metabolic rate (RMR) and on serum T3 levels in obese children and to investigate whether RMR changes are related to T3 changes. Sixty-four healthy, overweight, children (age: 12.1+/-1.1 years, body mass index 29.3+/-4.3 kg/m2) were studied during a 6-week weight reduction programme. RMR (by indirect calorimetry) total T3, total T4, TSH and fat-free mass (FFM) (by anthropometry) were measured at baseline and after 6 weeks of dietary treatment. Weight loss resulted in a 10.1% decline in RMR (P < 0.01) and a 23.4% decrease in serum T3 levels (P < 0.001). RMR was correlated with FFM before (r = 0.78, P < 0.001) and after weight loss (r = 0.76, P < 0.001). The changes in RMR were positively correlated with the changes in FFM (r = 0.48, P < 0.05) but also with the changes in serum T3 levels (r = 0.47, P < 0.05). The initial T3 levels predicted the subsequent fall in T3 that occurred after 6 weeks of dietary treatment (r = -0.60, P < 0.001). CONCLUSIONS: A significant decrease in serum T3 concentrations and resting metabolic rate occurred as a result of a 6-week weight reduction programme in an obese child population. The decline in T3 levels combined with fat-free mass loss could be responsible for the reduction in resting metabolic rate. PMID- 10378390 TI - Growth hormone treatment in a child with Williams-Beuren syndrome: a case report. AB - Growth retardation is a consistent finding in Williams-Beuren syndrome. The cause of short stature in this syndrome is unknown. Endocrine studies have failed to reveal abnormalities in the growth hormone-insulin-like growth factor I axis. We report a boy with confirmed Williams-Beuren syndrome, who was found to have classical growth hormone deficiency and responded well to growth hormone therapy. CONCLUSION: Although growth hormone deficiency is not likely to be a common cause of short stature in Williams-Beuren syndrome, we nevertheless recommend evaluation of the growth hormone-insulin-like growth factor I axis in all cases. PMID- 10378391 TI - Efficacy and safety of acarbose in patients with cystic fibrosis and impaired glucose tolerance. AB - Impaired glucose tolerance (IGT) is an increasingly frequent complication of cystic fibrosis (CF). In CF patients, a fast postprandial rise in plasma glucose is typically followed by a delayed but prolonged insulin response. Patients may develop symptoms of both hyper- and hypoglycaemia. The alpha-glucosidase inhibitor, acarbose, delays the hydrolysis and subsequent absorption of ingested carbohydrates. The aim of this study was to investigate the efficacy of acarbose in CF patients with IGT. During a 2-week inpatient period for treatment of Pseudomonas infection, 12 CF patients with IGT were studied in a double-blinded, randomized crossover trial. Each patient received acarbose (50 mg t.i.d.) for 5 days and placebo for 5 days (days 3-8 and days 10-14, respectively). Glucose, insulin and C-peptide responses to a standardized nutritional load were measured at baseline and at the end of each study period (Days 2, 8 and 14). Treatment with acarbose was associated with significant reductions in the mean value, mean peak values and the area under the curve of plasma glucose, insulin and C peptide, compared to respective baseline values and placebo. Gastro-intestinal disturbances were recorded in 67% of patients during therapy with acarbose. CONCLUSION: Acarbose has a positive therapeutic effect on glucose tolerance in cystic fibrosis patients, as shown by attenuation of postprandial plasma glucose increase and a significant decrease in insulin secretion response. However, acarbose treatment was associated with adverse gastro-intestinal effects that may prevent patients from accepting long-term therapy. PMID- 10378392 TI - Gastro-intestinal bleeding caused by leiomyoma of the small intestine in a child with neurofibromatosis. AB - Gastro-intestinal bleeding is an uncommon presentation in children with neurofibromatosis. Gastro-intestinal involvement caused by jejunal leiomyoma has only been described in adults. To the best of our knowledge, this is the first paediatric case of jejunal leiomyoma associated with neurofibromatosis. We present a 10-year-old girl with a 9-month history of anaemia and low gastro intestinal bleeding. Abdominal sonography and small bowel series showed a submucosal mass in the proximal jejunum. On surgery, a submucosal tumour was excised and histological examination suggested a diagnosis of "smooth muscle tumour of undetermined malignant potential". There were no recurrence of symptoms for 4 years after the operation. CONCLUSION: Jejunal leiomyoma should be considered in a child with neurofibromatosis presenting with gastro-intestinal bleeding. PMID- 10378393 TI - In vitro immunoglobulin response of fetal B-cells is influenced by perinatal infections and antibiotic treatment: a study in preterm infants. AB - The aim of our study was to analyse the influence of perinatal infections and administration of antibiotics on B-cell activity in blood cell cultures of preterm infants. We studied spontaneous and Escherichia coli induced immunoglobulin (Ig) secretion in 148 infants of 24 to 36 weeks of gestation: 53 healthy infants (Group I), 40 healthy infants receiving prophylactically antibiotics (Group II), 14 infants with intra-uterine infection (Group III) and 41 with nosocomial infection (Group IV). Spontaneous Ig secretion was significantly lower in neonates with intra-uterine infection (Group III) than in healthy infants of Group I. Nosocomial infections in Group IV increased spontaneous Ig synthesis, but only in the first days after birth. E. coli stimulation of peripheral blood mononuclear cells significantly increased Ig synthesis in healthy infants of Group I, whereas induced minimal Ig production in infected infants of Groups III and IV. Antibiotics given as prevention to Group II decreased Ig production in cell cultures as compared to healthy infants (Group I). CONCLUSION: The results indicate that perinatal infections and administration of antibiotics depress immunoglobulin secretion in cell cultures. We suggest that in vivo B-cell activity in infected preterm infants, and infants prophylactically receiving antibiotics, could also be depressed and result in decreased immunoglobulin production in these infants. PMID- 10378394 TI - Lack of lymphoid cell apoptosis in the pathogenesis of tonsillar hypertrophy as compared to recurrent tonsillitis. AB - The pathogenic mechanism of tonsillar hypertrophy is unknown and lacks a proper infectious or immunological explanation. Epidemiological studies point to polluted environments as the main cause of tonsillar hypertrophy in the adaptation of the juvenile organism. Tonsils and adenoids of 67 children aged 2 16 years (mean 5.9 years) were divided into three groups: recurrent tonsillitis (n = 21), recurrent tonsillitis with tonsillar hypertrophy (n = 21) and tonsillar hypertrophy without history of tonsillitis (n = 25). The following biological markers were studied: anti-streptolysin O antibody and anti-deoxy ribonuclease B antibody serology, microbiology and cell count of granulocytes in tonsils and adenoids as well as lymphocyte subsets and "ex vivo" endonuclease activity in tonsils. Anti-streptolysin O antibody and anti-deoxyribonuclease B antibody titres were significantly raised in recurrent tonsillitis. Positive bacterial cultures for Streptococcus pyogenes were rare in cases of tonsillar hypertrophy. T-lymphocytes counts were lower and the proportion of basophils was higher in hypertrophic tonsils than in recurrent tonsillitis. Two parameters of apoptosis were studied; the activation of endonuclease, inducing breakdown of DNA resulting in cell death, and the sensitivity to thapsigargin, known to trigger the cleavage of DNA by apoptotic endonuclease. In children with tonsillar hypertrophy both parameters were decreased contrasting with those with recurrent tonsillitis where apoptosis is increased. It may be speculated that the increase of basophils in children with tonsillar hypertrophy results in increased release of interleukin 4, which could prevent lymphoid apoptosis and lead to cell proliferation in tonsillar tissue. CONCLUSION: Whereas recurrent tonsillitis is characterised by apoptotic death of lymphoid tissue, tonsillar hypertrophy is caused by environmental pollution agents that trigger the chronic inflammatory process without apoptotic cell death. PMID- 10378395 TI - Congenital hydrocephalus internus and aqueduct stenosis: aetiology and implications for genetic counselling. AB - Genetic counselling in families with congenital hydrocephalus internus (CHI) in combination with aqueduct stenosis (AS) is often difficult due to an uncertain aetiology. We present a series of 35 patients with CHI and AS focusing on the aetiology and presumed recurrence risk for siblings. In 13 patients (37.1%) a genetic aetiology was identified with an increased recurrence risk for siblings. The relative frequency of patients with X-linked hydrocephalus in our sample was in accordance with the literature (2/35), but was more frequent in other diseases with Mendelian inheritance. CONCLUSION: In addition to the well-known X-linked and autosomal recessive forms of aqueduct stenosis with hydrocephalus, this malformation can occur in other diseases with Mendelian inheritance. This finding is of considerable importance for genetic counselling and prognosis. PMID- 10378396 TI - A mild variant of Desbuquois dysplasia. AB - On the basis of three newly observed cases (a pair of siblings and a sporadic case) and one previously reported case, we describe the clinical and radiological phenotype of a skeletal dysplasia resembling Desbuquois dysplasia. The skeletal alterations in the present disorder, including generalized osteopenia, mild modification of the vertebral endplates, epiphyseal flattening of the long bones, broad proximal femora with a spur-like projection of the lesser trochanters (a monkey wrench appearance of the proximal femora), and advanced carpal skeletal age, are almost identical to those of Desbuquois dysplasia. However, postnatal growth failure and minor spondylo-articular problems in the present disorder contrast with the conspicuous prenatal growth failure and severe spondylo articular deformities of Desbuquois dysplasia. Short stature in the present disorder does not reach the degree of Desbuquois dysplasia. Molecular investigation of one patient excluded abnormalities of the diastrophic dysplasia sulphate transporter gene. CONCLUSION: The combination of skeletal alterations identical to those of Desbuquois dysplasia with milder short stature and spondylo articular problems in the present patients suggests the nosological proposal of "a mild variant of Desbuquois dysplasia". PMID- 10378397 TI - A patent ductus arteriosus is associated with reduced middle cerebral artery blood flow velocity. AB - The study objective was to determine the effect of a patent ductus arteriosus (PDA) on cerebral blood flow velocity in ventilated, very low birth weight neonates, in the first 5 days of life. Sonography of the right middle cerebral artery and ductus arteriosus was performed using a colour Doppler technique. Statistical analysis was by stepwise regression. Thirty-one neonates without and 43 with a PDA, mean (SD) birth weight 1004 g +/- 192 and 1071 g +/- 227 respectively, were studied. The end diastolic and mean velocities were reduced (P = 0.008 and P = 0.129) and the resistive index was increased (P = 0.047) by a PDA. pH was inversely related to end diastolic and mean velocities (P = 0.015 and P = 0.003), suggesting that low pH may increase cerebral artery blood flow velocity. CONCLUSION: A patent ductus arteriosus reduces middle cerebral artery blood flow velocity in very low birth weight neonates. PMID- 10378398 TI - Substance misuse in early pregnancy and relationship to fetal outcome. AB - To establish the frequency of substance misuse in early pregnancy in an urban UK population, 807 consecutive positive pregnancy test urine samples were screened for a range of drugs, including cotinine as an indicator of maternal smoking habits. A positive test for cannabinoids was found in 117 (14.5%) samples. Smaller numbers of samples were positive for other drugs:- opiates (11), benzodiazepines (4), cocaine (3) and one each for amphetamines and methadone. Polydrug use was detected in nine individuals. Only two samples tested positive for ethanol. The proportion with a urine cotinine level indicative of active smoking was 34.3%. The outcome of the pregnancy was traced for 288 subjects. Cannabis use was associated with a lower gestational age at delivery (P < 0.005), an increased risk of prematurity (P < 0.02) and reduction in birth weight (P < 0.002). Whilst maternal smoking was associated with a reduction in infant birth weight (P < 0.05), this was less pronounced than the effect of other substance misuse. CONCLUSION: This study suggests that one in six women in South London are using drugs in early pregnancy and that cannabinoid use is associated with a poorer pregnancy outcome. PMID- 10378399 TI - Chronic respiratory morbidity following premature delivery--prediction by prolonged respiratory support requirement? AB - Neonatal chronic lung disease (CLD) is usually diagnosed if an infant remains oxygen dependent beyond 36 weeks postconceptional age (PCA). Our aim was to determine whether a shorter duration of respiratory support accurately predicted subsequent respiratory morbidity. A total of 103 infants, median gestational age 29 weeks (range 23-35), were followed prospectively for 5 years. They had a birth weight of < 1500 g or, if a birth weight of between 1500 and 2000 g, had required neonatal ventilatory support. Parents completed diary cards; their child had positive symptom status if, in any one year, they coughed and/or wheezed on at least 3 days per week for a 4-week period or for at least 3 days following each upper respiratory tract infection. Subsequent respiratory morbidity, positive symptom status in years 1 and 2 or all 5 pre-school years, was related to various definitions of prolonged respiratory support: intermittent positive pressure ventilation dependence > 7 days; oxygen dependence > 28 days and oxygen dependence > 36 weeks PCA. In years 1 and 2, 25 children were symptomatic and 22 in all 5 years. The patients with subsequent respiratory morbidity were distinguished from those without by requiring longer respiratory support (P < 0.05). Logistic regression analysis demonstrated only oxygen dependence beyond 28 days was independently related to subsequent respiratory morbidity (P < 0.01). The positive predictive values and likelihood ratios (95% confidence intervals) for positive symptom status in all 5 years were for intermittent positive pressure ventilation > 7 days 35% (16-53) and 19.5 (1.01-3.76), for oxygen dependency > 28 days 42% (23-61) and 2.20 (1.45-5.02) and for oxygen dependency >36 weeks PCA 35% (13-58) and 1.67 (0.65-4.31). CONCLUSION: Oxygen dependency at 28 days of age remains a useful criterion on which to diagnose "neonatal" chronic lung disease. PMID- 10378400 TI - Granulocyte colony-stimulating factor receptor expression on neutrophils of term and preterm neonates with and without signs of infection. AB - Neutrophils are an essential component of the human host defence system against infection. Recombinant human granulocyte colony-stimulating factor induces neutrophilia and enhances effector functions of mature neutrophils. Since the biological effects of granulocyte colony-stimulating factor (G-CSF) are mediated by its receptor, we investigated the expression of G-CSF receptor on the surface of neutrophils of term and preterm neonates (n = 22) with and without signs of infection and of healthy adults (n = 13) by flow cytometry. In healthy adults, the percentage of neutrophils expressing G-CSF receptor was higher compared to cord blood of term and preterm neonates (87% vs 53%, P < 0.05). Between 2 and 32 h of life, neonates with signs of infection showed lower values of G-CSF receptor expression compared to neonates without signs of infection (32% vs 54%, P < 0.05). No correlation was detectable between expression of G-CSF receptor and gestational age. CONCLUSION: Expression of granulocyte colony-stimulating factor receptor on neutrophils is lower than in adults. This may adversely affect granulopoiesis and neutrophil function during the neonatal period. Moreover, granulocyte colony-stimulating factor receptor expression seems to be down regulated during neonatal infection. PMID- 10378401 TI - Iodine deficiency in Turkey. AB - Turkey is an iodine deficiency area. The overall goitre prevalence is thought to be 30%, and most epidemiological studies give figures compatible with mild to moderate iodine deficiency. However, it is suspected that there are regions where iodine deficiency might be more severe than previously known. In this study the goitre prevalence and iodine status in a mountain village in Central Anatolia were investigated and the results compared to those of an urban area with mild iodine deficiency. Parameters of iodine status in the mountainous region showed severe iodine deficiency comparable to that in Central Africa. It seems that there are regions in Turkey where current programmes of salt iodization will be inadequate to correct the problem of iodine deficiency. CONCLUSIONS: Our observations suggest that regional variations in iodine status may impede the success of salt iodization programmes, which alone may not be adequate for correction of the problem country-wide. Alternative sources of iodine should be considered in addition to expanded and more efficient salt iodization programmes. PMID- 10378402 TI - Axial and peripheral bone mineral acquisition: a 3-year longitudinal study in Chinese adolescents. AB - We performed a 3-year longitudinal study of a group of 179 healthy Chinese adolescents (92 boys and 87 girls) aged from 12 to 16 years to determine the effects of puberty, physical activity, physical fitness, and calcium intake on the acquisition of bone mass. At yearly intervals for 3 consecutive years we recorded nutrition, calcium intake and anthropometric measurements, and assessed pubertal status according to Tanner. Bone mass of the lumbar spine was determined by dual-energy X-ray absorptiometry and radial bone mass by single-photon absorptiometry. Physical fitness and level of physical activity were assessed and muscle strength and power determined by isokinetic testing. Peripheral bone mass correlated with axial skeleton bone mass. Age, pubertal staging, physical fitness and muscle strength were significantly associated with bone mass increments on cross-sectional univariate and regression analysis. Longitudinal regression analysis showed that the most important factor affecting bone mass accretion in adolescents in both sexes was their pubertal stage. In boys, bone mass increment throughout the study was greater in children who were already in the advanced pubertal stages on entering the study than in those who started puberty in year 2 or 3 of the study. The percentage change in bone mineral content of the forearm and in bone mineral density of the lumbar spine was greater than 25% in the advanced pubertal group as compared to around 20% in the less mature group. For girls, the reverse was true. The increment of bone mass during the study period was significantly greater in those who presented in the earlier pubertal stages than in those who were at the more advanced stage of puberty on entry into the study. There was no significant effect of calcium intake and physical activities on the bone mass accretion. CONCLUSION: In Chinese adolescents, bone mineral accretion at adolescence is not influenced by exercise, level of physical fitness and calcium intake. In both sexes, and especially in girls, to optimally increase bone mass, regular physical exercise programmes should be instituted well before the onset of puberty rather than at or after it. Once puberty starts, these interventions may have no or only limited effect. PMID- 10378403 TI - Congenital alveolar proteinosis caused by a novel mutation of the surfactant protein B gene and misalignment of lung vessels in consanguineous kindred infants. AB - Congenital alveolar proteinosis and misalignment of lung vessels are rare disorders. We report on five infants of consanguineous kindred. All infants were delivered at term after uneventful pregnancies. Shortly after birth they developed respiratory failure and severe persistent pulmonary hypertension. All died despite intensive care. Lung tissue of two infants was studied. Histological examination revealed combination of alveolar proteinosis and misalignment of lung vessels in one patient, alveolar proteinosis in the other. Immunostaining demonstrated surfactant protein B (SP-B) deficiency in both patients' lungs. In a further sibling, analysis of broncho-alveolar lavage fluid showed decreased surfactant protein. PCR and direct sequence analysis of the SP-B gene revealed three novel mutations. One of them, a single base deletion, shifts the reading frame at amino acid 122 and creates a premature termination of translation in exon 6. No mature SP-B protein is produced. CONCLUSION: Surfactant protein B deficiency caused by mutations of the respective gene and misalignment of lung vessels can concur. Both diseases may have a pathogenetic factor in common. PMID- 10378404 TI - The necessity of building population specific prediction equations for clinical assessment of pulmonary function tests. AB - Fitting adequate prediction equations for pulmonary function test (PFT) parameters is crucial in the analysis of lung function tests and their interpretation. Our work aimed at studying the necessity of building population specific prediction equations, rather than using prediction equations built-in in commercial equipment. We used as an example results of studies carried out among Israeli schoolchildren. Second to sixth grade children (7-13 years old), 1064 boys and 1211 girls, were studied in Tel-Aviv. PFT (forced vital capacity, forced expiratory volume in 1st second, peak expiratory flow, forced expiratory flow in 50% volume, forced expiratory flow in 75% volume) performed by these children were adjusted for height, weight and age, for each sex separately, by a multiple regression procedure. Predicted PFT parameters of 300 boys and 301 girls aged 7 13 years, living along the southern shore of Israel, were calculated using the equations built for the same aged Tel-Aviv children as well as the prediction equations built-in in the spirometer used. The ratios between the observed PFT parameters in the southern children and their expected values, using the Israeli population specific equations, were around 1.00. Using the built-in equations resulted in ratios around 0.90. CONCLUSION: The development of population specific prediction equations for PFT parameters is necessary. Such equations should be used both in clinical assessment to minimize misclassification (healthy/sick child) and in epidemiological studies. PMID- 10378405 TI - Anaemia, hypoproteinaemia and abdominal distension. PMID- 10378407 TI - Phototherapy sequela in a child with congenital erythropoietic porphyria. PMID- 10378406 TI - Low erythrocyte docosahexaenoic acid in malnourished, often breast-fed, Pakistani infants: a matter of concern? PMID- 10378408 TI - 18q-syndrome with coeliac disease. PMID- 10378409 TI - Increased serum levels of soluble adhesion molecules in young children with atopic dermatitis. PMID- 10378410 TI - Cutaneous leishmaniasis: response to cryotherapy treatment. PMID- 10378411 TI - Trypanosoma cruzi and human ubiquitin are immunologically distinct proteins despite only three amino acid difference in their primary sequence. AB - The high similarity between Trypanosoma cruzi and human ubiquitin prompted us to characterize the human humoral immunity to host and parasite ubiquitin in Chagas disease and its possible role in Chagas autoimmunity. We have used a simplified one step purification procedure to partially purify T. cruzi ubiquitin. Using this preparation we have performed ELISA and Western blots, to show that chagasic sera recognise T. cruzi but not human or Leishmania ubiquitin indicating a species-specific response. Our results show that despite the high degree of similarity in the primary structure of human and T. cruzi ubiquitins, the three amino acid difference is sufficient to distinguish parasite versus host proteins. PMID- 10378412 TI - Antigen-specific response of murine immune system toward a yeast beta-glucan preparation, zymosan. AB - Zymosan, a particulate beta-glucan preparation from Saccharomyces cerevisiae, shows various biological activities, including anti-tumor activity. We have previously shown that soluble beta-glucan initiated anti-tumor activity was long lived and was effective even by prophylactic treatment at 1 month prior to tumor challenge. However, the activity by zymosan was relatively short-lived. Antigen specific responses of mice to zymosan might be a causative mechanism. In this paper, mice were immunized with zymosan and antibody production and antigen specific responses of lymphocytes to zymosan were analyzed. Sera of zymosan immune mice contained zymosan-specific IgG assessed by enzyme-linked immunosorbent assay and FACS. Spleen and bone marrow cells of zymosan-immune mice showed higher cytokine production in response to zymosan. Specificity of zymosan specific responses were also analyzed using various derivatives prepared from zymosan. These facts strongly suggested that mice recognize zymosan as antigen in addition to non-specific immune stimulant. PMID- 10378413 TI - Production and partial characterization of anti-cord factor (trehalose-6,6' dimycolate) IgG antibody in rabbits recognizing mycolic acid subclasses of Mycobacterium tuberculosis or Mycobacterium avium. AB - An ELISA with cord factor (trehalose-6,6'-dimycolate) is useful for the serodiagnosis of tuberculosis. To clarify the exact antigenic epitope in cord factor, recognized by a rabbit anti-cord factor IgG antibody, and to ascertain the most sensitive and specific diagnostic test antigen, rabbits were immunized with two kinds of cord factors isolated from Mycobacterium tuberculosis or Mycobacterium avium and the reactivities of the sera were tested against cord factors or the component mycolic acid methyl esters by ELISA. The serum from rabbits immunized with M. tuberculosis cord factor was highly reactive against M. tuberculosis cord factor, but less reactive against M. avium cord factor. In contrast, the serum from rabbits immunized with M. avium cord factor was highly reactive against M. avium cord factor but less reactive against M. tuberculosis cord factor. Moreover, the serum from rabbits immunized with M. tuberculosis cord factor reacted against mycolic acid methyl esters, especially methoxy mycolic acid methyl ester. On the other hand, the serum from rabbits immunized with M. tuberculosis cord factor was less reactive against trehalose-6-monomycolate and not reactive against sulfolipid (2,3,6,6'-tetraacyl trehalose 2'-sulfate). From these results, it was concluded that the anti-cord factor IgG antibody, produced experimentally in rabbits, recognized the differences in the cord factor structures, i.e. the hydrophobic moiety rather than the carbohydrate moiety. It was also noted that the serum from rabbits immunized with M. tuberculosis cord factor was highly reactive against methoxy mycolic acid as an epitope. This paper is the first to describe how the anti-cord factor IgG antibody can recognize the mycolic acid subclasses, which differ according to the species of mycobacteria. PMID- 10378415 TI - Chronic gastrointestinal inflammation. AB - Chronic gastrointestinal inflammation is one of the most common types of inflammatory process which affects humans. It is diverse in aetiology, pathogenesis and manifestation. There are also features of chronic inflammation at different sites within the gastrointestinal tract which provide a common thread in terms of the approaches which may be used in investigating these intriguing processes. This paper provides an overview of the mucosal changes in chronic gastrointestinal inflammation. Conserved and variable features of inflammation at different sites extending from the oral cavity to the rectum are highlighted. The involvement of different inflammatory cell types within any diagnostic entity is considered and the progression from an acute to chronic inflammatory condition explored. Important issues in the maintenance of a chronic inflammatory state are the balance between pro- and anti-inflammatory pressures, the driving force behind the inflammation and immune response that is occurring and the mechanisms for curtailment of unwanted or harmful responses which may damage the host. Thus inflammation is likely to result when there is persistence of a driving force and/or imbalance in the pro- and anti-inflammatory mechanisms in the tissue involved. PMID- 10378414 TI - Human T cell recognition of the Mycobacterium leprae LSR antigen: epitopes and HLA restriction. AB - We have in this work mapped epitopes and HLA molecules used in human T cell recognition of the Mycobacterium leprae LSR protein antigen. HLA typed healthy subjects immunized with heat killed M. leprae were used as donors to establish antigen reactive CD4+ T cell lines which were screened for proliferative responses against overlapping synthetic peptides covering the C-terminal part of the antigen sequence. By using this approach we were able to identify two epitope regions represented by peptide 2 (aa 29-40) and peptide 6 (aa 49-60), of which the former was mapped in detail by defining the N- and C-terminal amino acid positions necessary for T cell recognition of the core epitope. MHC restriction analysis showed that peptide 2 was presented to T cells by allogeneic cells coexpressing HLA-DR4 and DRw53 or DR7 and DRw53. In contrast, peptide 6 was presented to T cells only in the context of HLA-DR5 molecules. In conclusion, the M. leprae LSR protein antigen can be recognized by human T cells in the context of multiple HLA-DR molecules, of which none are reported to be associated with the susceptibility to develop leprosy. The results obtained are in support of using the LSR antigen in subunit vaccine design. PMID- 10378416 TI - Characterization of the respiratory chain of Helicobacter pylori. AB - The respiratory chain of Helicobacter pylori has been investigated. The total insensitivity of activities of NADH dehydrogenase to rotenone and of NADH cytochrome c reductase to antimycin is indicative of the absence of the classical complex I of the electron transfer chain in this bacterium. NADPH-dependent respiration was significantly stronger than NADH-dependent respiration, indicating that this is a major respiratory electron donor in H. pylori. Fumarate and malonate exhibited a concentration-dependent inhibitory effect on the activity of succinate dehydrogenase. The activity of succinate-cytochrome c reductase was inhibited by antimycin, implying the presence of a classical pathway from complex II to complex III in this bacterium. The presence of NADH fumarate reductase (FRD) was demonstrated in H. pylori and fumarate could reduce H2O2 production from NADH, indicating fumarate to be an endogenous substrate for accepting electrons from NADH. The activity of NADH-FRD was inhibited by 2 thenoyltrifluoroacetone. A tentative scheme for the electron transfer pathway in H. pylori is proposed, which may be helpful in clarifying the pathogenesis of H. pylori and in opening new lines for chemotherapy against this bacterium. PMID- 10378417 TI - pH-dependent binding of Helicobacter pylori to pig gastric mucins. AB - A microtiter-based assay was developed to study the binding of Helicobacter pylori to pig gastric mucins purified by density-gradient centrifugation in CsCl/4 M guanidinium chloride. Binding of H. pylori was observed over the 'mucin' band as well as with 'low-density' components in the gradients, and binding to the latter was more pronounced when incubations were performed at 37 degrees C as compared to 20 degrees C. At a lower pH, binding of H. pylori (strain SVA 40) to the 'high-density' mucins from pig antrum was increased but binding to the 'low density' ones was decreased. Binding of the P466 strain (Le(b)-specific) was mainly associated with the 'mucin' band, whereas the MO19 strain reacted preferentially with the 'low-density' components. In summary, H. pylori may bind to gastric mucins and the binding is influenced by temperature, pH and the repertoire of bacterial adhesins. PMID- 10378419 TI - Susceptibility in vitro of Helicobacter pylori to cetylpyridinium chloride. AB - The antimicrobial agent cetylpyridinium chloride (CPC) which is used in therapy of oro-pharyngeal infections and for antiseptic treatment of the oral cavity is active against different bacterial species. Determination of the minimal inhibitory concentration (MIC) using the agar dilution technique revealed that the gastric pathogen Helicobacter pylori in vitro is highly susceptible to CPC as indicated by an MIC of 10 microM (3.4 microg ml(-1)) which was significantly lower than the MIC of CPC against other bacterial species, which were analyzed in comparison to H. pylori. Bacteria of the genus Campylobacter, various Streptococcus spp., Staphylococcus aureus and Escherichia coli showed higher MICs ranging from 100 microM to 2 mM. In summary, this finding renders CPC-containing drugs candidates possibly useful for eradication or for the prevention of transmission of the gastric pathogen. PMID- 10378418 TI - Metronidazole resistance and virulence factors in Helicobacter pylori as markers for treatment failure in a paediatric population. AB - The eradication rate obtained using the classical triple therapy containing metronidazole, amoxicillin and bismuth citrate, was determined in 57 paediatric patients with digestive disorders, according to the susceptibility to metronidazole of the Helicobacter pylori strains (determined by agar dilution) and the cagA and vacA status (determined by PCR). Eradication was obtained in 38 out of 43 patients (88.3%) infected by H. pylori with metronidazole MIC < or = 2 mg l(-1), in 3 out of 6 patients (50%) when MIC was 4-8 mg l(-1) and in 4 out of 8 patients (50%) when MIC was > 8 mg l(-1). Among patients infected with cagA+ and cagA- strains an eradication rate of 75% (6/8) and 75% (18/24) was found, and 50% (3/6) and 80% (21/26) among vacA s1- and vacA s2-infected subjects (P > 0.05). H. pylori eradication depends on the susceptibility of the strain to metronidazole, being higher in patients infected with susceptible H. pylori. However, according to our data the cagA or vacA status was not an important factor in treatment failure in the eradication of H. pylori. PMID- 10378420 TI - Molecular methods for typing of Helicobacter pylori and their applications. AB - Microbial typing is a useful tool in clinical epidemiology for defining the source and route of infection, for studying the persistence and reinfection rates, clonal selection in the host and bacterial evolution. Phenotypic methods such as biotyping, serotyping and hemagglutinin typing have little discriminatory power compared to genotypic methods concerning the typing of Helicobacter pylori. Therefore great efforts have been made to establish useful molecular typing methods. In this context, the most frequently used genotypic methods are described based on our own experience and the literature: (1) restriction endonuclease analysis, (2) endonuclease analysis using pulsed-field gel electrophoresis, (3) ribotyping, (4) polymerase chain reaction (using either random primers or repetitive DNA sequence primers), and (5) polymerase chain reaction-restriction fragment length polymorphism analysis of e.g. the urease genes. Furthermore, reproducibility, discriminatory power, ease of performance and interpretation, cost and toxic procedures of each method are assessed. To date no direct comparison of all the molecular typing methods described has been performed in the same study with the same H. pylori strains. However, PCR analysis of the urease gene directly on suspensions of H. pylori or gastric biopsy material seems to be useful for routine use and applicable in specific epidemiological situations. PMID- 10378421 TI - Helicobacter pylori detection in human biopsies: a competitive PCR assay with internal control reveals false results. AB - A polymerase chain reaction assay (PCR) for the diagnosis of Helicobacter pylori in human gastric biopsies was developed. To prevent false-negative results while performing PCR on human tissues, an internal control is necessary. Primer set ACT1-ACT2 which specifically amplifies a 542-bp fragment of the 16S rRNA gene of H. pylori was used. dUTP and hot-start were used to prevent false-positives from carryover of previous products and avoid non-specific extension products. A competitive internal control DNA fragment was constructed to detect the presence of inhibitors. Biopsies from 101 unselected patients with gastric symptoms were tested. PCR results were compared with results from microscopy of histological sections and conventional culturing for H. pylori. Forty-two percent of the biopsies were found to contain compounds inhibiting the PCR. The addition of the internal control assures the performance of the PCR assay and is an important quality control parameter. PMID- 10378422 TI - The effect of Helicobacter pylori on neutrophil chemotaxis is independent of cagA. AB - Sixteen Helicobacter pylori strains were studied in order to determine their neutrophil chemotactic activity and the association with the presence cagA gene. Neutrophil chemotactic activity was detected by a modified Boyden chamber method and the results were expressed in terms of chemotactic index (CI). The presence of cagA was determined by PCR. Of the 16 strains, eight were cagA+ and eight were cagA-. All of the isolated strains showed chemotactic activity. The mean value of CI of the patient group was significantly higher than the negative control (P < 0.01). The mean value of CI of zymosan-activated serum (P < 0.05) and the reference strain H. pylori NCTC 11637 (HP11637) (P < 0.01) was significantly higher than the patient group's mean value of CI. There were no statistical significance in the CI between cagA+ and cagA- strains (P > 0.05). It is concluded that H. pylori attracts neutrophils by chemotaxis, however, there is no association with cagA. PMID- 10378423 TI - cagA gene and vacA alleles in Spanish Helicobacter pylori clinical isolates from patients of different ages. AB - The prevalence of the cagA gene and vacA alleles in 124 Spanish Helicobacter pylori clinical isolates from patients of different ages ranging from 3 to 78 years was studied (21 patients < or = 10 years, 30 patients 11-20 years, 17 patients 21-40 years, 31 patients 41-60 years and 25 patients 61-80 years). The cagA gene and vacA s1 or vacA s2 alleles were identified by PCR from the strain. 66.9% of the isolates were cagA+ and 33.1% cagA-. vacA s1 was detected in 48.4% of the isolates and vacA s2 in 51.6%. 44.4% of patients were cagA+/vacA s1, 22.5% were cagA+/vacA s2, 4% were cagA-/vacA s1 and 29% were cagA-/vacA s2. The percentage of cagA+ isolates and the vacA s1 alleles in the different groups were as follows: 23.8% and 28.6% in 0-10 years, 40% and 30% in 11-20 years, 88.2% and 70.6% in 21-40 years, 90.3% and 70.9% in 41-60 years and 92% and 44% in the 61-78 years group. 93% (54/58) of isolates found in ulcer patients and 90.9% (10/11) of isolates from gastritis patients older than 20 years were cagA+. In patients younger than 20 years ulcer disease was rare with 60% of isolates being cagA+ (3/5) compared with 31.6% cagA+ isolates (12/38) in patients suffering from gastritis in the younger group. The prevalence of the cagA gene and vacA s1 allele increased with age, being more frequent in older patients than in younger. PMID- 10378424 TI - Protection against Helicobacter pylori infection by intestinal immunisation with a 50/52-kDa subunit protein. AB - A mouse model of Helicobacter pylori infection was used to evaluate the vaccine antigen potential of the citrate synthase homologue protein purified from the H. pylori NCTC 11637 strain. Mice were immunised with the protein by intra-Peyer's patch immunisation. This route gives maximal intestinal immunisation and was used to screen oral vaccine candidate antigens without the added complication of simultaneously testing oral delivery systems. Two weeks post-immunisation mice were infected with Sydney strain H. pylori and 4 weeks after infection the mice were killed and the level of H. pylori infection in the stomach determined. Pre immunisation with the 50/52-kDa protein led to a 84-91% reduction in H. pylori infection compared to unimmunised controls. PMID- 10378425 TI - IgG antibodies to Lewis type 2 antigens in serum of H. pylori-infected and noninfected blood donors of different Lewis(a,b) blood-group phenotype. AB - Individuals of the Le(b+)/secretor phenotype revealed a stronger natural immune response to Le(x) and Le(y) epitopes irrespective of Helicobacter pylori serologic status. In contrast, H. pylori-infected Le(b-) type individuals showed a significantly higher proportion of strong responders to Le(x) antigen compared with the H. pylori-uninfected subgroup. The data suggest that the immune response to Lewis type 2 determinants is related to both the H. pylori serologic status and the Le(a,b) phenotype of the host. PMID- 10378426 TI - Changes in the mucosal expression of interleukin 15 in Helicobacter pylori associated gastritis. AB - Transcripts for interleukin (IL) 15 were detected in the gastric mucosal samples of 5/5 (100%) patients with no evidence of Helicobacter pylori infection and in 4/14 (28%) H. pylori-infected patients (P< 0.05). Both IL-15 mRNA and IL-15 protein were detected in 1/6 (17%) patients who successfully underwent H. pylori eradication therapy, before treatment and in 5/6 (83%) cases after eradication. Even though a parallel significant (P < 0.03) improvement of gastritis score occurred after eradication, the severity of gastritis did not differ according to the mucosal IL-15 expression among H. pylori-infected patients, irrespective of the CagA serology. This study demonstrates, for the first time, that transcripts for IL-15 are expressed in the human gastric mucosa. Changes occurring during H. pylori colonisation and after eradication raise the hypothesis that H. pylori may down-regulate IL-15 expression in the gastric mucosa. PMID- 10378427 TI - Impact of ELISA and immunoblot as diagnostic tools one year after eradication of Helicobacter pylori in a multicentre treatment study. AB - The performance of serological tests for Helicobacter pylori infections is hampered by the persistence of antibodies after eradication therapy or spontaneous healing. Detection of different antigens or immunoglobulin classes might have an impact on the validity of serodiagnosis. The aim of this study was to assess the decrease in IgA and IgG antibody levels after eradication of H. pylori. Serum samples of 242 patients with active duodenal ulcer were tested with the ELISA and the immunoblot (IB) techniques for H. pylori-specific IgA and IgG antibodies before therapy and 1 year after successful eradication. From a total of 81 patients paired sera were available. At the end of the follow-up period ELISA antibody titres from the IgA class had decreased from a mean value of 6.69 to 4.26 units (P = 0.0001), and IgG class antibody titres from a mean value of 21.9 to 12.1 units (P = 0.0001). Regarding seroreversion, from 34 initially IgA positive sera 16 (47%), and from 74 IgG positive sera 18 (24%), had definitively reverted to 'negative'. One year after eradication, when tested with the immunoblot, the antibody responses against specific antigens of 37% IgA-positive sera (23/62) and 8% IgG-positive sera (6/78) reverted to 'negative', compared to a seroreversion rate of 27% of the anti-CagA IgA-positive sera (18/67) and of 9% of the anti-CagA IgG-positive sera (7/79). In conclusion, despite an overall significant decrease of H. pylori antibodies, both tests cannot be recommended for monitoring treatment success. PMID- 10378428 TI - Animal models for gastric Helicobacter immunology and vaccine studies. AB - Over the last decade animal models have been used extensively to investigate disease processes and therapy for Helicobacter pylori infections. The H. pylori animal models which have been used in pathogenesis and vaccine studies include the gnotobiotic pig, non-human primates, cats, dogs, and several species of rodents including mice, rats, gerbils and guinea pigs. H. felis infection of mice and H. mustelae infection of ferrets have also been used. Recently, investigators have begun using transgenic mice and gene-targeted 'knock-out' mice to investigate Helicobacter infections. Each of these animal models has distinct advantages and disadvantages which are discussed in this minireview. The choice of an animal model is dictated by factors such as cost and an understanding of how each model will or will not allow fulfillment of experimental objectives. PMID- 10378429 TI - 13C-1H dipolar recoupling under very fast magic-angle spinning using virtual pulses. AB - A new solid-state NMR pulse sequence for recoupling 13C-1H dipolar interactions under magic-angle spinning is proposed, which works under a spinning speed of a few to several tens kilohertz. The sequence is composed of two different frequency switched Lee-Goldburg sequences, and the modulation of the spin part of the 13C-1H dipolar interaction is introduced by a virtual pulse sequence consisting of unitary operators connecting the rotating frame and the tilted rotating frame. When the cycle time of the spinning is equal to or twice the cycle time of the sequence, the 13C-1H dipolar interactions can be recoupled. The sequence is insensitive to experimental imperfections such as rf inhomogeneity or frequency offset, and the resulting lineshape can be represented by a simple analytical equation based on the zeroth-order average Hamiltonian. Experimental results for [2-(13)C] L-valine x HCl are reported. PMID- 10378430 TI - NMR study of monomethylammonium cation in (CH3NH3)5Bi2Cl11 ferroelectric polycrystal. AB - Spin-lattice relaxation times T1 and T1p are determined for protons in three polycrystals (CH3NH3)5Bi2Cl11, (CD3NH3)5Bi2Cl11 and (CH3ND3)5Bi2Cl11. The temperature dependencies of the relaxation times obtained for (CH3NH3)5Bi2Cl11 and (CD3NH3)5Bi2Cl11 are interpreted as a result of correlated motions of the three-proton groups of the monomethylammonium cation. The minimum of the T1p relaxation time is explained as a result of the oscillations of the symmetry axis of the whole cation. PMID- 10378431 TI - 19F/29Si distance determination in fluoride-containing octadecasil by Hartmann Hahn cross-polarization under fast magic-angle spinning. AB - 19F/29Si Hartmann-Hahn continuous wave cross-polarization (CP) has been applied under fast magic-angle spinning (MAS) to a powder sample of octadecasil. Strong oscillations occur during CP on a sideband matching condition between the isolated 29Si-19F spin pairs formed by the silicons in the D4R units and the fluoride anions. The magnitude of the dipolar coupling constant was deduced directly from the line-splitting between the intense singularities of the Pake like patterns obtained by Fourier transformation of the oscillatory polarization transfer. The corresponding Si-F internuclear distance, r = 2.62 +/- 0.05 A, is found to be in very good agreement with the X-ray crystal structure and the value of 2.69 +/- 0.04 A recently reported from rotational echo double resonance (REDOR) and transferred echo double resonance (TEDOR) nuclear magnetic resonance (NMR) experiments. Furthermore, the CP technique is still reliable under fast MAS where both REDOR and TEDOR sequences suffer from severe artefacts due to finite pulse lengths. In octadecasil, a spinning frequency of approximately 14 kHz is shown to be necessary for an effective suppression of 19F-19F spin diffusion. The influences of experimental missettings and radiofrequency (RF) field inhomogeneity are taken into account. PMID- 10378432 TI - Solid-state photodimerization of 4-aryl-1,4-dihydropyridines studied by 13C CPMAS NMR spectroscopy. AB - 13C CPMAS NMR spectroscopy has been applied to monitor the solid-state reaction of two different photodimerizing 4-phenyl-1,4-dihydropyridines yielding a cage dimer in one case and an anti-dimer in the other case. The spectra of the reacting monomers exhibit a magnetical inequivalence of chemically equivalent CO and C2/4 carbon atoms caused by a rotation of the pseudoaxially oriented 4-phenyl substituent out off the plane through N1, C3, C8 which could be determined by X ray crystal structure analyses of the centrosymmetrically arranged monomers. The 13C CPMAS NMR monitoring of the cage dimer formation proves that the reaction takes place in two steps via a syn-dimer for which a non-symmetrical structure was derived from the spectrum. The non-symmetrical structure was confirmed by X ray crystal structure analysis of one structurally related derivative. A centrosymmetric structure for both the finally formed cage dimer and the anti dimer of the other monitored photoreaction was proved by their spectra with one set of signals for each half of the dimers. respectively. Thus, conformational properties of the molecules as well as the symmetry of the products can be directly derived from the 13C CPMAS NMR spectra. PMID- 10378433 TI - Off-angle correlation spectroscopy applied to spin-1/2 and quadrupolar nuclei. AB - A two-dimensional correlation experiment is described, in which homonuclear dipolar couplings are used to realize through-space magnetization exchange on spin-1/2 (31P) and on quadrupolar nuclei (23Na and 11B). In the detection period, Magic Angle Spinning is applied to enhance resolution, and the dipole couplings are re-introduced in the mixing period by spinning off the Magic Angle. The dependency of the exchange rates on the mixing time and the spinning angle is investigated. The influence of strong spin-locking during mixing is discussed, and shown in the spin-1/2 case to remove the dependence on chemical shift offset effects. For quadrupolar spins, the experiment yields information on the relative tensor orientations of the coupled quadrupoles. Applications to crystalline sodium aluminum diphosphate, sodium sulphite, and potassium borate glasses are shown. PMID- 10378434 TI - Sodium-23 MAS NMR study on nuclear quadrupole interaction in NA(1-x)Ag(x)NO2. AB - Ag-impurity effects on the first- and second-order quadrupole interaction (QI) at 23Na site in an isomorphic mixed system, Na(1-x)Ag(x)NO2 (x = 0, 0.0084, 0.026, 0.079, 0.094, 0.16), have been investigated by employing 23Na (I = 3/2) magic angle spinning nuclear magnetic resonance (MAS NMR) technique. The central transition (CT) and satellite transition (ST) are simultaneously observed with this system. From the spectral analysis, the quadrupole parameter and its distribution width are obtained as a function of Ag concentration. From the intensity loss of CT MAS centerband and of the envelope function of ST MAS sidebands due to impurities, the range of their influence on the second- and first-order QI is estimated. The estimated ranges contain the second and first neighbouring Na sites from the resonating 23Na nucleus for the first- and second order QI, respectively. PMID- 10378435 TI - Application of 29Si and 27Al magic angle spinning nuclear magnetic resonance to studies of the building materials of historical monuments. AB - We report the application of 29Si and 27Al magic angle spinning nuclear magnetic resonance (MAS NMR) studies on building stones from historical monuments to obtain direct information about the degree of degradation and to observe the changes in the consolidated material after treatment with tetraethoxysilane (TEOS). Using the data obtained from deteriorated materials, a diagnostic laboratory, a suitable treatment and recommendations for building conservation may infer. A case study is presented using stones from Guanajuato City Main Church (Central Mexico). X-ray diffraction patterns to characterize the species present in the stones agree with the solid state NMR results. PMID- 10378436 TI - The effect of stimulus discriminability on stimulus-preceding negativities prior to instructive and feedback stimuli. AB - Stimulus-preceding negativity (SPN) was recorded in time estimation tasks that allowed the pre-instruction SPN and the pre-feedback SPN to be compared. In the task in which an acoustic tone was presented 3 s after a voluntary movement, (a) the level of stimulus discriminability (easy and difficult), and (b) the information content of the acoustic tone (feedback and instruction) were manipulated. The pre-instruction SPN over the right hemisphere tended to be larger under the difficult than under the easy level of discriminability, but the difference was only marginally significant. In contrast, the pre-feedback SPN over the right hemisphere was significantly larger under the easy than under the difficult level of discriminability. These findings suggest that the level of stimulus discriminability influences the pre-instruction and the pre-feedback SPN differently, and that it is probable that the pre-feedback and the pre instruction SPN do not have the same functional significance. PMID- 10378437 TI - Self-report of circadian type reflects the phase of the melatonin rhythm. AB - This study examined the relationship between circadian rhythm characteristics of the pineal hormone melatonin and individual differences in circadian type and mood. 95 healthy young men and 22 women were assessed each hour (00:00-07:00 h) for blood levels of melatonin throughout one night in the laboratory. Each subject was assessed for circadian type (morning, afternoon, or evening type) and morning mood (PANAS). Circadian type was strongly related to the melatonin acrophase but not to amplitude or time of year of assessment. Also, morning types evidenced a more rapid decline in melatonin levels after the peak than did evening types. Evening types were younger than were morning types. Female morning types reported more positive affect upon waking than did female afternoon or evening types. Males showed no such discrimination. Age was related to both melatonin acrophase and circadian type but did not explain the relationship between them. The results replicate and extend findings on circadian type and psychological and physiological variables. PMID- 10378438 TI - The effect of intention to learn novel, environmental sounds on the novelty P3 and old/new recognition memory. AB - Brain electrical activity was recorded while 32 young adults listened to frequent tones, infrequent target tones, and infrequent novel environmental sounds (some of which repeated). Subjects in the incidental group were not informed about the novel sounds, while subjects in the intentional group were informed and, in addition, were asked to memorize them for a subsequent memory test. Following the novelty oddball task, all subjects were given an old/new sound recognition memory task. The novelty P3 showed a more anterior scalp distribution and an effect of repetition in the incidental compared to the intentional group, suggesting that pre-categorizing the novel sounds influenced how they were processed. For the intentional group only, a subsequent memory effect was elicited by novel sounds that were subsequently recognized compared to those that were not, although both groups showed robust old/new ERP effects during the recognition task. The robust subsequent memory effects in the intentional group were associated with greater recognition accuracy of familiar environmental sounds relative to the incidental group. These data suggest that the encoding activity reflected in the ERP subsequent memory effect may have engendered an advantage in subsequent recognition memory performance. PMID- 10378439 TI - Evaluation of an adaptive automation system using three EEG indices with a visual tracking task. AB - A system was evaluated for use in adaptive automation using two experiments with electroencephalogram (EEG) indices based on the beta, alpha, and theta bandwidths. Subjects performed a compensatory tracking task while their EEG was recorded and converted to one of three engagement indices: beta/(alpha + theta), beta/alpha, or 1/alpha. In experiment one, the tracking task was switched between manual and automatic modes depending on whether the subject's engagement index was increasing or decreasing under a positive or negative feedback condition. Subjects were run for three consecutive 16-min trials. In experiment two, the task was switched depending on whether the absolute level of the engagement index for the subject was above or below baseline levels. It was hypothesized that negative feedback would produce more switches between manual and automatic modes, and that the beta/(alpha + theta) index would be most effective. The results confirmed these hypotheses. Tracking performance was better under negative feedback in both experiments; also, the use of absolute levels of engagement in experiment two resulted in better performance. There were no systematic changes in these effects over three 16-min trials. The implications for the use of such systems for adaptive automation are discussed. PMID- 10378440 TI - Fuzzy diagnosis. PMID- 10378441 TI - A fuzzy clustering based segmentation system as support to diagnosis in medical imaging. AB - In medical imaging uncertainty is widely present in data, because of the noise in acquisition and of the partial volume effects originating from the low resolution of sensors. In particular, borders between tissues are not exactly defined and memberships in the boundary regions are intrinsically fuzzy. Therefore, computer assisted unsupervised fuzzy clustering methods turn out to be particularly suitable for handling a decision making process concerning segmentation of multimodal medical images. By using the possibilistic c-means algorithm as a refinement of a neural network based clustering algorithm named capture effect neural network, we developed the possibilistic neuro fuzzy c-means algorithm (PNFCM). In this paper the PNFCM has been applied to two different multimodal data sets and the results have been compared to those obtained by using the classical fuzzy c-means algorithm. Furthermore, a discussion is presented about the role of fuzzy clustering as a support to diagnosis in medical imaging. PMID- 10378442 TI - Obtaining interpretable fuzzy classification rules from medical data. AB - For many application problems classifiers can be used to support a decision making process. In some domains-in areas like medicine especially-it is preferable not to use black box approaches. The user should be able to understand the classifier and to evaluate its results. Fuzzy rule based classifiers are especially suitable, because they consist of simple linguistically interpretable rules and do not have some of the drawbacks of symbolic or crisp rule based classifiers. Classifiers must often be created from data by a learning process, because there is not enough expert knowledge to determine their parameters completely. A simple and convenient way to learn fuzzy classifiers from data is provided by neuro-fuzzy approaches. In this paper we discuss extensions to the learning algorithms of neuro-fuzzy classification (NEFCLASS), a neuro-fuzzy approach for data analysis that we have presented before. We present interactive strategies for pruning rules and variables from a trained classifier to enhance its readability, and demonstrate our approach on a small example. PMID- 10378443 TI - Fuzzy pre-processing of gold standards as applied to biomedical spectra classification. AB - Fuzzy gold standard adjustment is a novel fuzzy set theoretic pre-processing strategy that compensates for the possible imprecision of a well-established gold standard (reference test) by adjusting, if necessary, the class labels in the design set while maintaining the gold standard's discriminatory power. The adjusted gold standard incorporates robust within-class centroid information. This strategy was applied to biomedical data acquired from a MR spectrometer for the purpose of classifying human brain neoplasms. It is shown that consistent improvement (10-13%) to the discriminatory power of the underlying classifier is obtained when using this pre-processing strategy. PMID- 10378444 TI - Diagnostic monitoring in anaesthesia using fuzzy trend templates for matching temporal patterns. AB - A technique based on the concept of a 'fuzzy trend template' has been developed to identify characteristic patterns in multiple time-series. The method has its foundation in fuzzy logic and allows for the intuitive and transparent description of 'templates', which preserve nuances of vagueness, temporal relationships and quantitative descriptors. Evaluation of fuzzy trend templates can provide both belief and plausibility information for use in diagnostic applications. The technique has been applied to the diagnosis of specific problems in anaesthesia and has demonstrated sensitivity and specificity of 95 and 65%, respectively. Evaluation of fuzzy trend templates is, computationally, relatively efficient and has allowed a real-time implementation. The technique has the potential to be useful in any domain that requires temporal pattern recognition based on linguistic rules. PMID- 10378446 TI - Dichloroacetate (DCA) dosimetry: interpreting DCA-induced liver cancer dose response and the potential for DCA to contribute to trichloroethylene-induced liver cancer. AB - Pharmacokinetic studies with dichloroacetate (DCA) provide insights into the likelihood that trichloroethylene-induced liver cancers arise from formation of DCA as a metabolite and the mode of action by which DCA induces liver cancer. A simple physiologically based pharmacokinetic model was developed to analyze DCA blood concentration data from mice unexposed to or pre-treated with DCA. The large first pass metabolism of DCA in the liver is significantly reduced by DCA pretreatment. Because DCA inhibits its own metabolism, large increases in area under the blood concentration curve occur at lower doses than would be predicted from single-dose pharmacokinetic studies with naive mice. The dose metrics associated with the incidence of liver tumors in contrast to the multiplicity of tumors per animal may be different, suggesting potentially different roles in the cancer process for DCA versus its metabolites. By linking a model for trichloroethylene (TCE) pharmacokinetics with the DCA model, maximum levels of DCA potentially produced from TCE were estimated to be at or below the analytical chemistry detection limits. In addition, the predicted levels of DCA would be too small to produce the observed liver cancers following corn oil gavage exposure of mice to TCE. PMID- 10378445 TI - Effects of molybdate and pentachlorophenol on the sulfation of alpha-naphthol. AB - Sulfation is the conjugation of chemicals with sulfate, which usually decreases, but occasionally increases, their biological effects. The phenol-sulfotransferase inhibitor pentachlorophenol (PCP) is often used to distinguish the biological effects of a chemical from its sulfate conjugate. Recently, molybdate has been shown to decrease the hepatic concentration of 3'-phosphoadenosine 5' phosphosulfate (PAPS), the cosubstrate for sulfation. Therefore, the present study was designed to compare the effectiveness and specificity of molybdate and PCP as inhibitors of sulfation. Alpha-naphthol (125 and 250 micromol/kg, i.p.) was administered to rats and the sulfate and glucuronide conjugates excreted into urine were quantified for this comparison. Molybdate (5.0, 7.5, and 10 mmol/kg) decreased the 24-h cumulative urinary excretion of the sulfate conjugate of the lower dose of alpha-naphthol by 54, 53, and 55%, respectively, with corresponding compensatory increases in glucuronide excretion at the two lower doses of molybdate. PCP (20, 40, and 80 micromol/kg) similarly decreased the sulfation of alpha-naphthol by 48, 38, and 41%, respectively, but prevented compensatory increases in glucuronide excretion. Molybdate (2.5, 5.0, and 7.5 mmol/kg) decreased the sulfation of the higher dose of alpha-naphthol by 21, 30, and 44%, respectively, again with corresponding compensatory increases in glucuronide excretion. In contrast, PCP did not decrease significantly the sulfation of the higher dose of alpha-naphthol. These data suggest that molybdate is equally or more effective than PCP at inhibiting sulfation of alpha-naphthol, and appears to be more specific. PMID- 10378447 TI - Metabolites of acetaminophen trigger Ca2+ release from liver microsomes. AB - Release of mitochondrial calcium is believed to play a key role in the toxicity of acetaminophen in biological systems. Elevated cytosolic Ca2+ may also result from activation of calcium releasing channels. The major metabolites of acetaminophen, benzoquinone imine and 1,4-benzoquinone, induced Ca2+ release in isolated rat liver microsomes. The 1,4-benzoquinone-induced release of calcium was suppressed by ryanodine and fully inhibited by reduced glutathione. Concentrations of 1,4-benzoquinone that induced Ca2+ release did not affect the activity of the microsomal Ca2+, Mg2+-APTase. The binding of [3H]ryanodine to liver microsomes, however, was significantly decreased by 1,4-benzoquinone, suggesting a direct interaction of this metabolite with the ryanodine-binding protein (ryanodine receptor). These results suggest that cellular Ca2+ levels may be elevated by acetaminophen by pathways involving, in part, activation of Ca2+ releasing channels such as the ryanodine receptor. PMID- 10378448 TI - Chrysotile-mediated imbalance in the glutathione redox system in the development of pulmonary injury. AB - A significant depletion in the content of glutathione (GSH) and alteration in GSH redox system enzymes were observed in the lung of chrysotile-exposed animals (5 mg) during different developmental stages of asbestosis. In the alveolar macrophages (AM) of exposed animals, the depletion in GSH started from day 1 and reached a maximum at day 16, whereas in lung tissue the maximum depletion was observed when fibrosis has matured. It appears that cellular GSH depletion triggers oxidative stress in the system as observed from increased thiobarbituric acid reactive substance (TBARS) production and alteration in the activities of glutathione peroxidase (GPx), glutathione reductase (GR), glucose 6-phosphate dehydrogenase (G6PD) and glutathione S-transferase (GST), the enzymes regulating oxidative tone. The depletion in GSH was also observed in red blood cells (RBC) of the exposed animals reaching a maximum when fibrosis matured. Thus the observed depletion in GSH, ascorbic acid and alteration in GSH redox system enzymes may be involved in fibrosis and carcinogenesis induced by chrysotile. PMID- 10378449 TI - N-acetyl-L-cysteine protects against delta-aminolevulinic acid-induced 8 hydroxydeoxyguanosine formation. AB - 5-Aminolevulinic acid (ALA) is a heme precursor that accumulates in acute intermittent porphyria and lead poisoning. It has been shown that ALA induces free radical generation and may cause damage to proteins and DNA. In the present study, the effects of ALA on DNA damage and its prevention by N-acetyl-L-cysteine (NAC) and the antioxidant enzymes catalase (CAT) and superoxide dismutase (SOD) are investigated. Oxidative damage to DNA was quantitated by measuring the increase in 8-hydroxy-2'-deoxyguanosine (oh8dG) formation. The time-course study demonstrated that ALA causes a linear increase in oh8dG levels in Chinese hamster ovary (CHO) cells. However, direct lead exposure did not cause any measurable increase in oh8dG levels. In the presence of either NAC (1 mM) or antioxidant enzymes (10 u/ml SOD and 10 u/ml CAT), oh8dG levels returned to the corresponding control levels. This suggests a protective role for NAC and the antioxidant enzymes. To determine the effect of ALA on cell proliferation, cell numbers were counted at the end of 24 h of incubation in the presence and absence of ALA at different concentrations. Results showed that levels of ALA up to 5 mM do not inhibit cell proliferation. PMID- 10378450 TI - Influence of oral administration of a quaternary mixture of trihalomethanes on their blood kinetics in the rat. AB - Trihalomethanes (THMs; chloroform, bromoform, bromodichloromethane, dibromochloromethane), formed as by-products of chlorine disinfection, are found to occur in combination in drinking water supplies. THMs are metabolized by cytochromes P-450 and are likely substrates of CYP2E1. Therefore, it is possible that mixed exposure results in toxicokinetic interactions among THMs. The toxicokinetics of THMs during mixed exposures has not been investigated previously. The purpose of this study was to characterize the blood kinetics of the four THMs administered singly or in combination in the rat. A single dose of 0.25 mmol/kg or 0.5 mmol/kg b.w., of each THM alone, or of a quaternary mixture containing 0.25 mmol/kg of each THM (total dose of 1.0 mmol/kg) was administered by gavage. The venous blood concentrations of the THMs were measured by headspace gas chromatography (GC) at 20, 40, 60, 120, 180, 270 and 360 min post administration. Results showed a nonlinear relationship between the area under the blood concentration versus time curves (AUCs) and administered doses of THMs, suggesting that metabolism is saturated in this dose range. The venous blood concentrations of THMs following administration of the quaternary mixture were significantly higher compared to single exposures. The altered kinetics of THMs during combined exposures is consistent with the occurrence of mutual inhibition of their hepatic metabolism. Simulation exercises conducted with physiologically based toxicokinetic models support metabolic inhibition as the possible mechanism of the interaction among THMs. The data reported in this study provide the starting point for evaluating the significance of interactions among THMs in the risk assessment process. PMID- 10378451 TI - Cytotoxic versus genotoxic effects of nitric oxide (NO). AB - The pathobiochemistry of endogenous reactive nitrogen species includes functions in inflammation and carcinogenesis. Genotoxicity has been suggested to play a major role. Two donor compounds, spermine NONOate, which can release authentic nitric oxide (NO), and 3-Morpholino-sydnonimine hydrochloride (SIN-1), which generates NO together with superoxide, possibly yielding peroxynitrite (ONOO-), were investigated in L5178Y mouse lymphoma cells for cytotoxic and genotoxic effects. As demonstrated by cell growth, 'micronucleus' and 'comet' assays NO, with and without concomitant superoxide formation, did not induce significant genotoxicity at concentrations with low cytotoxicity. Therefore, at least for the three tested parameters and the chosen time window, the pronounced cytotoxicity exhibited by NO and its oxidative metabolites most likely outweighs any genotoxic potential. PMID- 10378452 TI - Influence of renal biomarker variability on the design and interpretation of occupational or environmental studies. AB - OBJECTIVES: To quantify and identify sources of within- and between-subject variability of microalbumin, N-acetyl-beta-D-glucosaminidase (NAG) and alanine aminopeptidase (AAP), three biomarkers used for early detection of renal injury, and to assess the consequences of this variability for the design and power of epidemiological studies. METHODS: Urinary excretion of microalbumin, NAG, AAP and creatinine as well as blood pressure (BP) were measured three times over a 2-year period among 142 healthy male workers. To minimise physiopathological and analytical sources of variation, standardised methods were used for urine collection and assays, and severe exclusion criteria were applied. At the first and third examinations, subjects completed the same questionnaire, providing information about their personal characteristics, tobacco and alcohol consumption, and health. A linear mixed model was used to estimate the within- and between-subject variance components and to analyse the relation between subjects' characteristics and the biomarkers. RESULTS: No change in the mean value of any of the biomarkers was observed over the 2-year period. Intra-class correlation coefficients between repeated measurements were 0.53, 0.57 and 0.56 for microalbumin, NAG and AAP, respectively; the between-subject variance was slightly higher than the within-subject variance. Subjects' age, BP, body mass index and smoking and drinking habits explained 7.2%, 12.5% and 4.2% of the total variance of microalbumin, NAG and AAP, respectively. CONCLUSIONS: In this healthy population of male workers, day-to-day differences in biomarker values appeared to be nearly as great as differences between subjects. The within-subject variance of these biomarkers is not high enough to justify systematic repeated measurements in epidemiological surveys. But, in some situations where the number of subjects is limited, measuring the subjects twice may improve study power by reducing the total variance by about 25% for each biomarker. Taking the above covariates into account would slightly improve study power and the accuracy of parameter estimates for NAG, but would add little to the analysis of microalbumin and AAP. PMID- 10378453 TI - Deferoxamine delays the development of the hepatotoxicity of acetaminophen in mice. AB - The hepatotoxicity of acetaminophen is conventionally ascribed to metabolism by CYP450 to N-acetyl-p-benzoquinone imine and covalent binding to proteins. We investigated a potential role for oxidative stress by determining the effect of the ferric chelator deferoxamine (Desferal) on acetaminophen (paracetamol) induced hepatotoxicity in mice. Administration of deferoxamine (75 mg/kg) 1 h after a toxic dose of acetaminophen (300 mg/kg) significantly delayed the development of the toxicity without altering covalent binding. In saline-treated mice serum ALT was 18 +/- 2 IU/l. In acetaminophen-treated mice serum alanine aminotransferase (ALT) was 779 +/- 271 at 2 h, 7421 +/- 552 IU/l at 4 h, 5732 +/- 523 IU/l at 8 h, and 5984 +/- 497 IU/l at 24 h. In acetaminophen plus deferoxamine-treated mice, serum ALT was 80 +/- 10 at 2 h, 472 +/- 74 IU/l at 4 h, 2149 +/- 597 IU/l at 8 h, and 5766 +/- 388 at 24 h. Deferoxamine at 1 h after acetaminophen did not decrease serum ALT at 12 h; however, deferoxamine at 1 and 4 h, or deferoxamine at 1 h plus N-acetylcysteine at 4 h to replete hepatic glutathione, decreased the toxicity from 5625 +/- 310 IU/l to 3436 +/- 546 IU/l and 3003 +/- 282 IU/l, respectively. Deferoxamine plus N-acetylcysteine at 1.25 h after acetaminophen was more effective at decreasing the 24 h toxicity than N acetylcysteine alone. In acetaminophen treated mice, higher doses of deferoxamine (150-300 mg/kg) at 1 h greatly increased the observed hepatotoxicity at 4 h in a dose responsive manner, but deferoxamine alone was nontoxic. PMID- 10378454 TI - Multiple origins of the mtDNA 9-bp deletion in populations of South India. AB - The origins and genetic affinities of the more than 500 tribal populations living in South Asia are widely disputed. This may reflect differential contributions that continental populations have made to tribal groups in South Asia. We assayed for the presence of the intergenic COII/tRNALys 9-bp deletion in human mtDNA in 646 individuals from 12 caste and 14 tribal populations of South India and compared them to individuals from Africa, Europe, and Asia. The 9-bp deletion is observed in four South Indian tribal populations, the Irula, Yanadi, Siddi, and Maria Gond, and in the Nicobarese. Length polymorphisms of the 9-bp motif are present in the Santal, Khonda Dora, and Jalari, all of whom live in a circumscribed region on the eastern Indian coast. Phylogenetic analyses of mtDNA control region sequence from individuals with the 9-bp deletion indicate that it has arisen independently in some Indian tribal populations. Other 9-bp deletion haplotypes are likely to be of Asian and African origin, implying multiple origins of the 9-bp deletion in South India. These results demonstrate varying genetic affinities of different South Indian tribes to continental populations and underscore the complex histories of the tribal populations living in South Asia. PMID- 10378455 TI - Genetic population structure of two African-Ecuadorian communities of Esmeraldas. AB - The genetic structure of two African-Ecuadorian communities, Rio Cayapas and Viche (Esmeraldas province, northwest Ecuador), was studied on the basis of ACP1, ADA, AK1, CA2, ESD, GLO1, G6PD, PGD, and PGM1 subtypes and thermostability, PGM2, HBbeta, F13A, F13B, ORM1, AHSG, C6, C7, and APOC2 gene frequency, and migration data on 255 individuals. The fixation index of Wright (F(ST)), correspondence, and genetic distance analysis were applied to compare the genetic relationships between these communities and other American populations of African ancestry. F(ST) values from the migration data and surname origins suggest that Rio Cayapas is genetically more isolated and shows less mobility and admixture than does Viche. The genetic admixture estimates indicate a large contribution of African genes to the gene pool of both communities (74.3% to 58.4%), whereas the proportion of the Amerindian component differs significantly (14.5% in Rio Cayapas to 27.6% in Viche). PMID- 10378456 TI - Differential contribution of indigenous men and women to the formation of an urban population in the Amazon region as revealed by mtDNA and Y-DNA. AB - The human populations of the Brazilian Amazon were formed by interethnic crosses between Europeans, Africans, and Amerindians. The relative contribution of men and women of different ethnic groups was not homogeneous, since the social policies of the first three centuries of Brazilian colonization encouraged mating between European men and indigenous women and, later on, African women. In order to test this model based on historical data, we compared the relative contribution of the Y-DNA and mtDNA of Amerindian and non-Amerindian populations to the formation of the urban population of the town of Belem, in the Amazon region, on the basis of a C-->T mutation at locus DYS199 present in 90% of the Amerindian Y-DNA and of five markers that define 99% of the mitochondrial sequences of Amerindians. The contribution of indigenous men to the formation of this population was less than 5%, whereas the contribution of indigenous women was estimated at more than 50% of the mitochondrial sequences of the same population. Thus, the present results demonstrate that the contribution of indigenous women to the formation of the Belem population was 10 times higher than the contribution of indigenous men, a genetic consequence of social behavior and attitudes of the past; our results also help clarify the process of integration of indigenous communities into the urban societies in Brazil and possibly in other countries. PMID- 10378457 TI - Beta-fibrinogen allele frequencies in Peruvian Quechua, a high-altitude native population. AB - Elevated hematocrits, which are found in many high-altitude populations, increase the oxygen-carrying capacity of blood and may represent an adaptation to hypoxic environments. However, as high hematocrit increases blood viscosity, which in turn is associated with hypertension and heart disease, it may be advantageous for high-altitude populations to limit other factors that contribute to increased blood viscosity. One such factor is the plasma concentration of the coagulation protein fibrinogen. Several common polymorphisms in the beta-fibrinogen gene have been identified that affect fibrinogen concentrations. We determined the allele frequencies of three of these polymorphisms (G/A-455(HaeIII), C/T-148(HindIII), and G/A+448(MnlI)) in sample groups drawn from three populations: Quechua speaking natives living at over 3,200 m in the Peruvian Andes, North American natives (Na-Dene) from coastal British Columbia, and Caucasian North Americans. The frequencies of the alleles previously shown to be associated with increased fibrinogen levels were so low in the Quechuas that their presence could be accounted for solely by genetic admixture with Caucasians. Frequencies in the Na Dene, a Native American group unrelated to the Quechua, were not significantly different from those in Caucasians. PMID- 10378458 TI - Growth patterns among seminomadic pastoralists (Datoga) of Tanzania. AB - Anthropometric measurements made on 470 individual children (age 0-18 years) from a seminomadic population of Datoga pastoralists living in northern Tanzania were used to describe patterns of child growth. Comparisons with reference growth curves derived from American samples suggest that pastoral Datoga children grow poorly in this region. Body compositional changes with age differed markedly from the reference population. There were negligible fat gains through childhood, even among females. Comparison with data on other East African pastoralists showed that population growth performance is intermediate between that of nomadic and settled pastoralists. Little catch-up growth occurs during childhood, and adolescence appears to be delayed among males. The results contribute to the growing database on health indicators for African pastoralists and suggest a need for further research to investigate mechanisms for growth stunting in these populations. PMID- 10378459 TI - Effects of loading on the biomechanical [correction of biochemical] behavior of molars of Homo, Pan, and Pongo. AB - In a previous study, we found systematic differences in the biomechanical behavior of modern human molars using finite element stress analyses (FESA), which led us to propose that molars are adapted to differently-directed loads depending on their position within the mouth (Spears and Macho [1998] Am. J. Phys. Anthropol. 106:467-482). While the FESA results thus derived have not been verified experimentally, such an interpretation seemed reasonable. To refine the model previously presented, this study assessed the effects of 1) food particle size on the biomechanical behavior of molars, and those of 2) differences in morphology, particularly enamel thickness, on stress distribution. In order to appraise the evolutionary significance of the findings, the FESA results for modern humans were subsequently compared with those obtained for molars of one individual of Pan and Pongo, respectively. Bearing in mind limitations imposed by the FESA models created and analyzed in this study, constant cleavage-type loads and cuspal tip loads at different directions were employed on all teeth: this facilitated comparisons of patterns of stress distribution across molars and species. In Pan and Homo, cleavage-type loads exerted by big food particles tended to be better dissipated anteriorly than posteriorly, although trends in Pongo were less clear-cut. Furthermore, similar to modern humans, the buccal cusps of mandibular molars appeared to be able to dissipate the loads associated with a pestle-type action, while maxillary molars were better designed to dissipate the loads which would result if they acted as mortars against which the food is crushed/ground. While increases in enamel thickness lowered the overall stress values in teeth only slightly, changes in outer morphology could have a more profound effect on these stress levels. Overall, Pan appeared to be most generalized, while Homo and Pongo showed a number of unique specializations, which are in accordance with what is currently understood about their respective masticatory apparatus and dietary niche. PMID- 10378460 TI - Fracture trauma in a medieval British farming village. AB - Farming is among the three most hazardous occupations in modern society and perhaps also held a similar position during the medieval period. The goal of this study was to determine if there is a significant difference in frequencies and patterns of longbone fracture trauma observed between rural and urban activity bases that distinguish farming as a particularly dangerous occupation during the medieval period. The longbones of 170 individuals excavated from Raunds, a rural medieval British site (10th-12th centuries AD) were examined for fractures and compared to data collected from four contemporary British medieval sites, one rural and three urban. The fracture frequency for the Raunds individuals (19.4%) was significantly different from the urban sites (4.7-5.5%). Female fractures were characterized by injury to the forearm, while the males were predisposed to diverse fracture locations. Clinical research provided a source of documented farm-related trauma from North America and Europe where the crops and animals raised, the manual chores performed, and the equipment used in traditional or small-scale farms have changed little in form or function since the medieval period. Nonmechanized causes of injury contribute to approximately 40% of all modern farm-related injuries and are attributed to falls from lofts and ladders, animal assaults and bites, and falls from moving vehicles. These hazardous situations were also present in the medieval period and may explain some of the fracture trauma from the rural sites. A high fracture frequency for both medieval males and females is significantly associated with farming subsistence when compared to craft-orientated urban dwellers. PMID- 10378461 TI - Sexing potential of fragmentary and pathological metacarpals. AB - The use of metacarpal dimensions for determining skeletal sex is investigated. Previous studies on sexual dimorphism in the metacarpals (Scheuer and Elkington [1993]; J. Forensic Sci. 38:769-778; Falsetti [1995]; J. Forensic Sci. 40:774 776; Smith [1996]; J. Forensic Sci. 41:469-477) used multiple variables, which limits the application potential of the methodology. Using six measurements for each metacarpal, I generated 35 linear discriminant functions based on expected taphonomic or pathological preservation scenarios. The number of variables per function ranged from 2-5. Normal, jackknifed, and cross-validation classification matrices indicated a sex prediction accuracy in the 79-85% range. MC4 produced the most consistent functions. Overall accuracy in the validation samples ranged from 75-90%. ANOVA and MANOVA analyses indicated that population effects are insignificant, which may allow for the application of the functions without knowledge of the ancestral background of the individual. This, combined with the variety of preservation scenarios considered, provides accurate sex estimators for incomplete individuals. However, the population specificity of the insignificant population group effects remains untested. PMID- 10378462 TI - Incidence and distribution of postcranial fractures in the prehistoric population of San Pedro de Atacama, Northern Chile. AB - Trauma incidence analysis in skeletal populations has been very popular among skeletal biologists during the last two decades. In this context, the work of Lovejoy and Heiple ([1981] Am. J. Phys. Anthropol. 55:529-541) has been quoted as a landmark because their analysis rested on a populational approach, avoiding simple assumptions about cause and etiology. In this study, we apply to the prehistoric population of San Pedro de Atacama, northern Chile, an approach similar to that carried out by Lovejoy and Heiple (1981). The results obtained point to a peak of risk of fractures among old people, estimated age around 45 years. The distribution of fractures by sex and age suggests that the prevailing etiology is related to accidents and not violence. When the frequencies of fractures are compared, the Libben population shows a much higher incidence than the Atacamenean population. It is suggested that this difference can be explained by peculiarities of the subsistence economies of the two populations. PMID- 10378463 TI - Spondyloarthropathy identified as the etiology of Nubian erosive arthritis. AB - Slight variation in manifestation of different diseases may allow a single individual with one disease to mimic the "classic" appearance of another, as evidenced by the frequent confusion of spondyloarthropathy with rheumatoid arthritis. Analysis of population occurrence of arthritis (rather than isolated skeletons) facilitates more precise diagnosis. Northeast Africans living around 2,000 years before present were clearly afflicted with a form of spondyloarthropathy. Lack of inclusion of spondyloarthropathy in the differential diagnosis of erosive arthritis led to past misclassification of Nubians as having rheumatoid arthritis. While evidence of spondyloarthropathy abounds in the literature of human skeletal disease, pre-Columbian Old World rheumatoid arthritis is still elusive. The current study further documents the absence of rheumatoid arthritis in Nubians, supporting the hypothesis that rheumatoid arthritis began in the New World. PMID- 10378464 TI - Brief communication: Resistance to Falciparum malaria in alpha-thalassemia, oxidative stress, and hemoglobin oxidation. AB - A recent survey conducted on Vanuatu Island suggests that resistance to Plasmodium falciparum in alpha-thalassemic individuals may have an immunological basis. This study is important since it seems to undermine the current idea that red-cell genetic defects give protection against falciparum malaria by reducing intraerythrocytic growth and development of the parasite. However, the mechanisms underlying these clinical and genetic observations are not yet fully understood. Based on a review of the relevant literature, we first show that the model based on the interaction between hemoglobin (Hb) and membrane components may provide a molecular basis for the involvement of the immune response in genetic adaptation to malaria. Second, we discuss the main evolutionary implications of the model. Finally, we suggest two approaches by which anthropological studies could provide a useful way of testing the model: 1) analysis of the interactions of malaria resistance genes with genetic polymorphisms which affect the erythrocyte redox status and 2) study of the antimalarial effects of natural products (introduced as a part of a diet or for traditional antimalarial therapy) capable of interfering with the Hb/membrane interaction. PMID- 10378465 TI - Male reproductive effects of solvent and fuel exposure during aircraft maintenance. AB - Few studies have addressed the effects of mixed, low-level exposures to complex mixtures on a man's reproductive potential. In this prospective study, each subject was evaluated before first exposure and at 15 and 30 weeks after exposures had begun. A total of 50 men working on aircraft maintenance at an Air Force installation were included in the study. In addition, eight unexposed men were concurrently sampled. Industrial hygiene (IH) sampling and expired breath samples were collected for jet fuel as measured by total napthas, benzene--a component of jet fuel, 1,1,1-trichloroethane, methyl ethyl ketone, xylenes, toluene, and methylene chloride. Sperm production, structure, and function (sperm concentration, sperm motion, viability, morphology, morphometrics, and stability of sperm chromatin) were evaluated. Exposures were low. All mean IH measures were below 6 ppm, which is less than 10% of the Occupational Safety and Health Administration standard for all chemicals except benzene. Sheet metal workers had the highest mean breath levels for both total solvents (24 ppb) and fuels (28.3 ppb). For most sperm measures, mean values remained in the normal range throughout the 30 weeks of exposure. When jobs were analyzed by exposure groups, some adverse changes were observed. The paint shop group had a significant decline in motility of 19.5% at 30 weeks. Internal dose measures, however, did not show a significant association with spermatogenic changes. PMID- 10378466 TI - Acrylamide causes preimplantation abnormalities in embryos and induces chromatin adducts in male germ cells of mice. AB - Acrylamide, a known male postmeiotic germ cell mutagen, caused a dose-dependent increase in the frequency of morphologic abnormalities in preimplantation embryos. Single-cell eggs, growth retardation, and blastomere lysis were detected after paternal treatment with acrylamide (10 to 50 mg/kg, 5 d). The major effects were seen at weeks 1 to 3 after male treatment, with the highest level of abnormalities at the first week (> 90% vs. 5% in control). The frequency of abnormal four-day embryos was similar to preimplantation loss assessed at 15 to 16 d p.c. A > 100-fold elevation of chromatin adducts in sperm was observed during 1st and 2nd week after treatment, after which adduct levels decreased to baseline level. However, morphologic defects in embryos are not fully explained by the spermatid adduct curve. These findings demonstrate the effects of paternal exposure to acrylamide on preimplantation development and indicate a potential risk to the offspring of men exposed to acrylamide. PMID- 10378467 TI - Effects of 2-bromopropane on spermatogenesis in the Sprague-Dawley rat. AB - In 1995, 2-bromopropane (2-BP) was associated with occupational reproductive and hematopoietic toxicity in Korea. The effect of 2-BP on spermatogenesis, or Leydig cells, has not been determined in adult rats. In the present study, 40 ten-week old Sprague-Dawley (SD) rats were treated orally with 3.5 g/kg/d of 2-BP for 3 consecutive days. At 1, 3, 5, 7, 14, 28, 42, and 70 d after treatment, testes were perfused with Karnovsky's solution or immersed in Bouin's solution, embedded in plastic or Epon and evaluated with light and electron microscopy. DNA ploidy distributions of testicular suspensions were determined by flow cytometry, which allowed comparison of quantitative spermatogenesis with histopathologic observations. Degeneration of spermatogonia was observed during Stages I-IV in seminiferous tubules on Day 1 after treatment. Spermatocytes, spermatids, Sertoli cells, and Leydig cells appeared normal in the early stage of the study. Whereas spermatid retention in Stages IX-XI was observed on Day 7 after treatment, depletion of spermatocytes and spermatids continued over time, followed by a marked increase of germ cells on Day 42 after treatment. However, the seminiferous tubules did not completely recover by study termination. Leydig cell cellularity increased mildly without any significant morphologic modification at the end of the study. Immunohistochemistry using an antibody against proliferating cell nuclear antigen (PCNA), showed an increased number of immunoreactive Leydig cells in the interstitium. In the flow cytometry analysis, proportions of diploid and tetraploid cells gradually decreased time-dependently until Day 28 after treatment, but showed an increase on Day 42, followed by a decrease on Day 70 after treatment. These data are strengthened by qualitative descriptions of lesions observed by histopathology. These results suggest that a high dose of 2-BP can decrease spermatogenesis by adversely affecting spermatogonia followed by depletion of spermatocytes, spermatids, and spermatozoa, with subsequent testicular atrophy. The atrophied testes may not regenerate completely. The number of Leydig cells may increase mildly with 10 weeks of recovery. PMID- 10378468 TI - Chlorambucil-induced postclosure exencephaly and axial skeletal abnormalities in rat fetuses. AB - Open neural tube defects (NTD) are reported to arise from failure of the embryonic neural folds to close or rupture of a previously closed neural tube. The critical period for dysmorphogenesis of the neural tube by either mechanism is of clinical importance. We had previously reported that single doses of cyclophosphamide (CPA), administered to pregnant rats resulted in reopening of the closed neural tube. The objective of the present study was to determine if other antineoplastic drugs also had similar ability to cause rupture of the closed neural tube. Therefore, single doses of chlorambucil dissolved in bicarbonate buffer were administered to Wistar rats on one of gestation days (GD) 11 through 14 (i.e., well after closure of the neural tube) and fetuses were examined on GD 20. It was observed that the window of susceptibility extended from GD 11 through 14 and that a single dose of 10 mg/kg of the drug was most effective. The affected fetuses had the prosencephalon and mesencephalon parts of the brain protruding through a wide opening of the cranial roof. The exposed brain tissue was hemorrhagic and covered by a thin, transparent and often porous membrane. The cranial vault was missing. Numerous malformations of the axial skeleton and of several nonneural organs were found to accompany the brain defect. Electron microscopic and light microscopic studies revealed extensive cell death, fragmentation, and phagocytosis of dead cells and vacuolization of the neuroepithelium (NE) within 12 h of the drug treatment. Cell death per se was not pronounced in the cranial mesenchyme (ME). The vacuoles in the NE coalesced into small cavities. The ME failed to proliferate adequately and organize itself into the cranial vault primordium. Hemorrhage and disorganization of the NE was progressive. The fluid that escaped into the interstitium seemed to extend into the cysts that developed in the ME externally and into the ventricular system internally. The edema of the ME thus might have contributed to disruption of precursor mesenchymal tissue and consequently to malformation of the cranial bones. Large hematomas and cysts also developed on the basal aspect of the crumbling brain vesicles and appeared to lift the neural tissue out of the shallow cranial cavity. As previously reported for CPA, these data provide evidence for the ability of chlorambucil to cause postclosure exencephaly in rat fetuses for a susceptible period that is considerably long. PMID- 10378469 TI - Pyridine nucleotide flux and glutathione oxidation in the cultured rat conceptus. AB - It is proposed that protection of the developing embryo from chemical and environmental insults that produces oxidative stress requires a proper glutathione (GSH) and pyridine nucleotide status in both the embryo and extra embryonic membranes. Modulation of pyridine nucleotide flux [NAD(H) and NAD(P)H] in the visceral yolk sac (VYS) by the thiol oxidants diamide and tert-butyl hydroperoxide (tBH) was studied in real time using microfiberoptic sensors in GD 10 rat conceptuses. Consecutive 5-min exposures to 125- and 250-microM diamide resulted in a fluorescence decrease of 14 and 32 Arbitrary Fluorescence Units (AFU). An additional consecutive exposure to 500-microM diamide caused an attenuated decrease followed by a rebound increase of 22 AFU. Consecutive 5-min exposures to tBH at 250 and 500 microM produced fluorescence decreases similar to that of 500 microM diamide, but the decreases were attenuated at 1000 microM. However, there was variability in the rebound increase. A 5-min exposure to tBH (500 microM) alone caused a fluorescence decrease of 14 AFU followed by a rebound increase of 8 AFU. The rate of fluorescence decrease was attenuated by 50% with pretreatment with the glutathione reductase (GSSG-Rd) inhibitor, BCNU (1,3, bis(2 chloroethyl)-1-nitrosourea), indicating that the decrease in surface fluorescence was probably attributable to a decrease in NADPH. Decreases in fluorescence, observed from the surface of the VYS, correlated with decreases in GSH/GSSG ratios in the embryos and the VYS. After exposure to tBH, GSH levels in conceptuses decreased at the end of 5 and 15 min, with a corresponding increase in oxidized glutathione (GSSG) at the end of 3, 5, and 15 min. Our results demonstrate that the increased production of GSSG on exposure to thiol oxidants correlates with a decrease in the reduced pyridine nucleotide, implying the presence of an active GSSG-Rd pathway in the conceptus during organogenesis, and implicating an important role of the pyridine nucleotides in the restoration of GSH homeostasis in the developing rat conceptus during organogenesis. PMID- 10378470 TI - Cadmium accumulation and effects on progesterone release by cultured human trophoblast cells. AB - This study was designed to examine the characteristics of cadmium bioaccumulation by human trophoblast cells in culture and the subsequent effect of cadmium exposure on progesterone production and syncytial formation. The accumulation of cadmium suggested a time- and dose-dependent relationship, although it was not significant. The rate of metal accumulation was similar in all cadmium-treated groups. After 72 h of continuous exposure to cadmium concentrations of 5, 10, and 20 microM, progesterone release was diminished to 69, 51, and 38% of control values (P < 0.05), respectively. When cells were exposed to cadmium from 72 to 96 h (after syncytial development), progesterone release exhibited the same pattern of decline in response to increasing cadmium concentrations. Histologic evaluation of whole mounts of trophoblast cells exposed to 20 microM CdCl2 for 96 h revealed that syncytial formation seemed to be uninhibited. The pattern of cadmium-accumulation by normal cultured human trophoblast cells suggests a time- and dose-relationship with a concomitant decrease in progesterone release that occurs without apparent inhibition of syncytial development. PMID- 10378471 TI - The conduct of a two-generation reproductive toxicity study via dermal exposure in the Sprague-Dawley rat--a case study with KBR 3023 (a prospective insect repellent). AB - KBR 3023, 1-(1-methyl-propoxycarbonyl)-2-(2-hydroxyethyl)-piperidine, a prospective insect repellent being developed by the Bayer Corporation, was evaluated for reproductive toxicity in the Sprague-Dawley rat. As the intended human use of the test compound is topical, the test system was also exposed to the compound via the dermal route. Specifically, the adult rats (P generation) were fitted with Elizabethan collars, to reduce the likelihood of oral ingestion, and dermally administered either 0, 50, 100, or 200 mg KBR 3023/kg body weight throughout the study (5 d/week) beginning at the onset of the 10-week premating period and continuing through the mating, gestation, and lactation phases. Clinical signs and changes in body weight and food consumption were assessed throughout the study. All adults and neonates underwent a gross necropsy examination. Tissues retained for microscopic examination from all adult animals included the kidney, liver, pituitary, reproductive organs, and samples of skin from the shaved dose site. In addition to the parameters noted above, the animals were evaluated for the effect of the test compound on estrous cycling, mating, fertility, gestation length, litter size, pup sex ratio, and pup viability. There were no test compound-related clinical signs or effects on body weight or food consumption observed in either the adults or the pups during any phase of the study. There were no compound-related effects on any reproductive or litter parameters. Dermal findings at the dose site (acanthosis and hyperkeratosis) were noted in both generations. Other than the dermal findings, no compound-related necropsy findings were seen in either the adults or the pups. No compound-related histopathologic findings were noted in the reproductive tissues of either the males or females. Based on these results, KBR 3023, administered as described in this study at dose levels as high as 200 mg/kg body weight (the physical limit of dermal application for this compound), did not demonstrate any reproductive toxicity. PMID- 10378472 TI - Offspring sex ratio as a monitor of the potential reproductive hazard of exposure to microwave radiation. PMID- 10378473 TI - The sex ratio of offspring of people exposed to boron. PMID- 10378474 TI - Effects of dietary vitamin E supplementation on pulmonary morphology and collagen deposition in amiodarone- and vehicle-treated hamsters. AB - Amiodarone (AM) is a potent antidysrhythmic agent that is limited in clinical use by its adverse effects, including potentially life-threatening AM-induced pulmonary toxicity (AIPT). The present study tested the ability of dietary supplementation with vitamin E (500 IU d,1-alpha-tocopherol acetate/kg chow) to protect against pulmonary damage following intratracheal administration of AM (1.83 micromol) to the male golden Syrian hamster. At 21 days post-dosing, animals treated with AM had increased lung hydroxyproline content and histological disease index values compared to control (P < 0.05), which were indicative of fibrosis. Dietary vitamin E supplementation for 6 weeks resulted in a 234% increase in lung vitamin E content at the time of AM dosing, and maintenance on the diet prevented AM-induced elevation of hydroxyproline content and disease index 21 days post-dosing. Dietary vitamin E supplementation also decreased hydroxyproline content and disease index values in hamsters treated intratracheally with distilled water, the AM vehicle. These results demonstrate a protective role for vitamin E in an in vivo model of AIPT, and suggest that this antioxidant may have non-specific antifibrotic effects in the lung. PMID- 10378475 TI - Impairment of TNF-alpha expression and secretion in primary rat liver cell cultures by acetaminophen treatment. AB - Tumor necrosis factor-alpha (TNF-alpha) is assumed to act as a mediator in toxic liver injury, aggravating the primary damage to the parenchymal liver cell, but also stimulating liver regeneration. Reports on the effect of acetaminophen in vivo on TNF-alpha transcript concentrations and serum TNF-alpha concentrations, under different experimental, or clinical conditions have yielded controversial results. We used primary rat hepatocyte and Kupffer cell cultures to test the direct action of subtoxic and toxic concentrations of acetaminophen on TNF-alpha expression and release. We observed a dose-dependent decrease of TNF-alpha mRNA in the hepatocytes, and of TNF-alpha release into the medium of hepatocyte cultures. The data also indicate an impairment of TNF-alpha release in Kupffer cell cultures after treatment with nontoxic, as well as with toxic, acetaminophen concentrations. The results suggest that inhibition of TNF-alpha expression and release in the liver is a consequence of acetaminophen exposure. It is at present unknown how this effect modulates the course of acetaminophen intoxication. PMID- 10378476 TI - Protection against chronic cadmium toxicity by caloric restriction. AB - Exposure to cadmium (Cd) can result in nephrotoxicity and osteotoxicity. Because Cd-induced nephrotoxicity involves oxidative stress and caloric restriction decreases oxidative stress, we examined whether reduced caloric intake will protect against Cd-induced nephrotoxicity. In addition, the protection against the osteotoxicity was also examined. Male and female Sprague-Dawley rats were provided drinking water containing 100 mg Cd/l. Since fluid intake relative to the body weight was higher in females as compared to the males, the Cd concentration in their water was reduced to 80 mg/l after 3 months and 65 mg/l after 6.5 months. During the 27 month exposure period the males and females consumed a total of about 5 g Cd/kg body weight. Food was restricted to 20 g/day after the first 3 months. During the unrestricted food intake period Cd exposure reduced the bone density in females by 23%, with a partial recovery and stabilization during the caloric restriction phase. Hepatic and renal Cd accumulation and corresponding metallothionein (MT) levels were very similar in both sexes. The reported critical Cd concentration for nephrotoxicity was reached by 9 months. Renal MT levels were maximum at this time. Despite a 1.5-fold increase in renal Cd concentration over the next 18 months, there was no significant increase in renal MT levels. In spite of high renal Cd levels and lack of availability of sufficient MT, there was no sign of nephrotoxicity, as measured by urinary protein and glucose excretion. It is concluded that caloric restriction prevents Cd-induced nephrotoxicity and also appears to control the osteotoxicity of Cd. PMID- 10378477 TI - Lead induced thiamine deficiency in the brain decreased the threshold of electroshock seizure in rat. AB - Many neurological disorders that occur frequently in lead intoxicated animals, have also been observed in thiamine deficient animals. To test whether lead intoxication could decrease the thiamine status and thresholds of electroshock seizure in rats, 3-week-old Wistar rats were treated with lead or lead plus thiamine. For comparison, a thiamine deficient group was included. Thiamine contents and transketolase activity, one of the thiamine dependent enzymes in the brain regions were significantly lowered by lead intoxication and thiamine deficiency. In both cases, thresholds of the electroshock seizure were significantly decreased. Thiamine supplementation reversed these signs and decreased the brain lead concentration in the lead treated group. The results from the present study suggest that the increased seizure susceptibility induced by lead intoxication in rats may be mediated at least in part through the changes of thiamine status. PMID- 10378478 TI - Antiestrogenicity of environmental polycyclic aromatic hydrocarbons in human breast cancer cells. AB - The total concentration of 14 polycyclic aromatic hydrocarbons (PAHs) was determined to be 3400-fold greater in a sediment sample from an industrial site on the St. Lawrence River (SLR), NY, than in a sediment sample from a non industrial site on the Kinderhook Creek (KC), NY. PAH fractions from extracts of the two environmental samples and two reconstituted mixtures as well as the 14 individual PAHs were examined for their toxic, estrogenic, and antiestrogenic activities using MCF-7 focus, recombinant human estrogen receptor (ER) binding, whole-cell ER binding, and 17beta-estradiol (E2) metabolism assays. PAH fractions from the KC and SLR were antiestrogenic; they significantly inhibited the formation of foci elicited in MCF-7 breast cancer cells by 1 nM E2. Eight of the 14 individual PAHs, and the reconstituted mixtures were also antiestrogenic. Results from the whole-cell ER binding assay and the radiometric analysis of E2 metabolism indicate that the PAHs detected in the KC and the SLR environmental samples induce antiestrogenic responses in metabolically intact human breast cancer cells through at least two mechanisms: one involving competition for the ER by a PAH metabolite and the other involving depletion of E2 through induction of metabolism. PMID- 10378479 TI - Acrolein toxicity: comparative in vitro study with lung slices and pneumocytes type II cell line from rats. AB - Toxicological effects of acrolein have been studied in precision-cut rat lung slices and in L2 cells, a rat pneumocyte II cell line. These two models were cultured for 24 h with or without acrolein (0-100 microM in L2 cells; 0-200 microM in lung slices). Treatment with this pneumotoxicant produced a concentration dependent decrease in intracellular ATP levels. Acrolein concentrations higher than 50 microM induced ATP decrease in slices, while this decrease occurred from 10 microM acrolein in L2 cells. Detoxification marker evaluations showed that mostly the glutathione pathway was altered after acrolein treatment in both models. Intracellular glutathione (GSH) levels were drastically increased with an acrolein concentration of 10 microM. This increase was concomitant with glutathione-S-transferase (GST) and glutathione reductase (GRED) activities in L2 cells. After this strong increase, these enzymatic activities as well as GSH levels were quickly decreased. In precision-cut rat lung slices, the induction of the glutathione pathway was less clear-cut. A bell-shaped dose response curve was observed with a maximum for 5 microM acrolein for GST and GRED activities. These differences between acrolein toxic ranges could be explained by the presence of an active detoxification pathway in slices compared to its relative lack in L2 cells. PMID- 10378480 TI - Inhibition of human erythrocyte Ca2+-ATPase by Zn2+. AB - Recent investigations suggest that Ca2(+)-ATPase from fish gills is very sensitive to Zn2+ (Hogstrand et al., 1996. Am. J. Physiol. 270, R1141-R1147). The effect of free Zn2+ ion on the human erythrocyte plasma membrane Ca2(+)-ATPase was investigated to explore the possible extension of this finding to humans. Membrane vesicles were prepared and the Ca2(+)-ATPase activity was measured as Ca2(+)-stimulated ATP hydrolysis and as ATP-dependent Ca2+ transport. The Zn2+ ion inhibited the erythrocyte Ca2(+)-ATPase by reducing Vmax and increasing the K0.5. While in the Ca2+ transport assay only the Vmax was affected at lower Zn2+ concentrations (50-100 pM), reduction of Vmax was always accompanied by an affinity decrease in the ATP hydrolysis assay. The Ca2(+)-ATPase was found to be inhibited by Zn2+ at extremely low concentrations. The IC10 and IC50 for Zn2+, at a Ca2+ concentration of 1.0 microM, were estimated at 4 and 80 pM, respectively. Although the Ca2(+)-ATPase might be more sensitive in vitro than in vivo conditions, the results suggest that physiological concentrations of Zn2+ may reduce the activity of the erythrocyte Ca2(+)-ATPase. Furthermore, disturbance of Ca homeostasis may be a mechanism causing Zn toxicity during exposure. PMID- 10378481 TI - Possible mechanism for lead inhibition of vascular endothelial cell proliferation: a lower response to basic fibroblast growth factor through inhibition of heparan sulfate synthesis. AB - Although lead inhibits the proliferation of vascular endothelial cells, the mechanism has been incompletely understood. A lower response to basic fibroblast growth factor (bFGF) of growing bovine aortic endothelial cells after exposure to lead was investigated using a cell culture system in the present study. It was shown that lead significantly decreased the incorporation of [3H]thymidine into the acid-insoluble fraction of the cells but the inhibition disappeared in the presence of bFGF neutralizing antibody. Pretreatment with lead resulted in a reduction of the stimulation by exogenous bFGF on the [3H]thymidine incorporation. Lead decreased endogenous bFGF bound to cell surface heparan sulfate proteoglycans in a concentration-dependent manner but not the high affinity FGF receptor without a change of the accumulation within the cells. In spite of such a change in the endogenous bFGF distribution, the total amount of the growth factor synthesized was not significantly changed by lead. Although the binding of [125I]bFGF to heparan sulfate proteoglycans can be directly inhibited by lead, the inhibition was not so marked. On the other hand, lead markedly suppressed the incorporation of [35S]sulfate into heparan sulfate accumulated in the cell layer and the conditioned medium, suggesting that the metal inhibited the synthesis of the glycosaminoglycan in growing endothelial cells. Inhibition of the [3H]thymidine incorporation by lead was significantly restored by heparin. Since the binding of bFGF to its receptor is strongly promoted by heparan sulfate, the present data suggest that lead inhibits vascular endothelial cell proliferation by induction of a lower response to endogenous bFGF through a suppression of heparan sulfate synthesis. PMID- 10378482 TI - Lead inhibits the core protein synthesis of a large heparan sulfate proteoglycan perlecan by proliferating vascular endothelial cells in culture. AB - We characterized proteoglycans synthesized by growing cultured bovine aortic endothelial cells after exposure to lead. Lead significantly decreased the incorporation of both [3H]glucosamine and [35S]sulfate into glycosaminoglycans accumulated in the cell layer and the conditioned medium of the cells in a dose dependent manner. Proteoglycans metabolically labeled with [35S]sulfate in the presence of lead were separated into heparan sulfate proteoglycans (HSPGs) and more highly charged chondroitin/dermatan sulfate proteoglycans by DEAE-Sephacel ion-exchange chromatography. It was found that lead markedly inhibited the synthesis of HSPGs. Sepharose CL-4B molecular sieve gel filtration showed that the marked decrease by lead occurred in the high molecular weight subclass of HSPGs. However, the length of heparan sulfate chains (approximately 50 kDa) was unchanged when analyzed by Sepharose CL-6B chromatography. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis of core proteins showed that lead reduced the accumulation of a high molecular weight (approximately 400 kDa) HSPG core protein in the cell layer and the conditioned medium; the core protein was identified as a perlecan core by Western blot analysis. It is suggested that lead inhibits the synthesis of the perlecan core protein in growing endothelial cells without a change of heparan sulfate chain length. The present data support the hypothesis that inhibition of endothelial cell proliferation by lead may result from a lower response of the cells to endogenous basic fibroblast growth factor whose binding to the receptor is strongly promoted by heparan sulfate chains of perlecan. PMID- 10378483 TI - In vitro suppressive effect of aflatoxin B1 on murine peritoneal macrophage functions. AB - We examined the immunosuppressive effects of aflatoxin B1 (AFB1), a toxic compound produced by the Aspergillus flavus, on murine peritoneal macrophages after in vitro pre-exposure. When thioglycollate-elicited macrophages pre-exposed to AFB1 were stimulated with lipopolysaccharide (LPS), antitumor activity induced by LPS was suppressed by 10 and 50 microM AFB1. In addition, the production of reactive intermediates including nitric oxide (NO), superoxide anion and hydrogen peroxide which have been known to be implicated in macrophage-mediated cytotoxicity, was decreased by AFB1 pretreatment in a dose-dependent manner. We also determined whether the macrophage-mediated cytokine production was altered by AFB1 in vitro pretreatment. AFB1 markedly inhibited TNF-alpha interleukin-1 (IL-1) and IL-6 production by LPS-stimulated macrophages. Taken together, these data indicate that AFB1 inhibits the killing ability of murine macrophages, decreases various secretory molecules in those cells and the macrophages would be one of many systems affected by AFB1. PMID- 10378484 TI - Functional outcome in amputation versus limb sparing of patients with lower extremity sarcoma: a matched case-control study. AB - OBJECTIVE: To quantify the differences in physical disability and handicap experienced by patients with lower extremity sarcoma who required amputation for their primary tumor as compared with those treated by limb-sparing surgery. DESIGN: Matched case-control study. Twelve patients with amputation were matched with 24 patients treated by limb-sparing surgery on the following variables: age, gender, length of follow-up, bone versus soft-tissue tumor, anatomic site, and treatment with adjuvant chemotherapy. PATIENTS: Patients who underwent above-knee amputation (AKA) or below-knee amputation (BKA) for primary soft-tissue or bone sarcoma, who had not developed local or systemic recurrence, and who had been followed up for at least 1 year since surgery. MAIN OUTCOME MEASURES: The Toronto Extremity Salvage Score (TESS), a measure of physical disability; the Shortform 36 (SF-36), a generic health status measure; and the Reintegration to Normal Living (RNL), a measure of handicap. RESULTS: Mean TESS score for the patients with amputations was 74.5 versus 85.1 for the limb-sparing patients. (p = .15). Only the physical function subscale of the SF-36 showed statistically significant differences, with means of 45 and 71.1 for the amputation versus limb-sparing groups, respectively (p = .03). The RNL for the amputation group was 84.4 versus 97 for the limb-sparing group (p = .05). Seven of the 12 patients with amputations experienced ongoing difficulty with the soft tissues overlying their stumps. CONCLUSIONS: There was a trend toward increased disability for those in the amputation group versus those in the limb-sparing group, with the amputation group showing significantly higher levels of handicap. These data suggest that the differences in disability between amputation and limb-sparing patients are smaller than anticipated. The differences may be more notable in measuring handicap. PMID- 10378485 TI - Nontraumatic spinal cord injury: incidence, epidemiology, and functional outcome. AB - OBJECTIVES: To identify and compare the incidence, demographics, neurologic presentation, and functional outcome of individuals with nontraumatic spinal cord injury (SCI) to individuals with traumatic SCI. DESIGN: A 5-year prospective study. SETTING: Level I trauma center of a Regional SCI Model System. PATIENTS: Two hundred twenty adult SCI admissions. MAIN OUTCOME MEASURES: Demographics, etiology, level and completeness of injury, Functional Independent Measure (FIM) scores. RESULTS: Of SCI admissions, 39% were nontraumatic in etiology (spinal stenosis, 54%; tumor, 26%). Compared to subjects with traumatic SCI, those individuals with nontraumatic SCI were significantly (p < .01) older and were more likely married, female, and retired. Injury characteristics revealed significantly more paraplegia and incomplete SCI within the nontraumatic SCI group (p < .01). Both nontraumatic and traumatic SCI individuals had significant FIM changes from rehabilitation admission to discharge (p < .01). Those with tetraplegia-incomplete nontraumatic SCI had significantly higher admission motor FIM scores and shorter rehabilitation length of stay than in the traumatic group (p < .05). Paraplegic-complete and paraplegic-incomplete nontraumatic SCI subjects had lower discharge motor FIM scores, FIM change, and FIM efficiency than those with traumatic SCI. Similar discharge-to-home rates were noted in both nontraumatic and traumatic SCI groups. CONCLUSIONS: These data suggest that individuals with nontraumatic SCI represent a significant proportion of SCI rehabilitation admissions and, although differing from those with traumatic SCI in demographic and injury patterns, can achieve similar functional outcomes. PMID- 10378487 TI - Measuring outcomes in children's rehabilitation: a decision protocol. AB - OBJECTIVE: To develop and test the feasibility and clinical utility of a computerized self-directed software program designed to enable service providers in children's rehabilitation to make decisions about the most appropriate outcome measures to use in client and program evaluation. DESIGN: A before-and-after design was used to test the feasibility and initial impact of the decision-making outcome software in improving knowledge and use of clinical outcome measures. SETTING: A children's rehabilitation center in a city of 50,000. PARTICIPANTS: All service providers in the children's rehabilitation center. Disciplines represented included early childhood education, occupational therapy, physical therapy, speech and language pathology, audiology, social work, and psychology. INTERVENTION: Using a conceptual framework based on the International Classification of Impairment, Disability, and Handicap (ICIDH), an outcome measurement decision-making protocol was developed. The decision-making protocol was computerized in an educational software program with an attached database of critically appraised measures. Participants learned about outcome measures through the program and selected outcome measures that met their specifications. The computer software was tested for feasibility in the children's rehabilitation center for 6 months. OUTCOME MEASURES: Knowledge and use of clinical outcome measures were determined before and after the feasibility testing using a survey of all service providers currently at the centre and audits of 30 randomly selected rehabilitation records (at pretest, posttest, and follow-up). RESULTS: Service providers indicated that the outcomes software was easy to follow and believed that the use of the ICIDH framework helped them in making decisions about selecting outcome measures. Results of the survey indicated that there were significant changes in the service providers' level of comfort with selecting measures and knowing what measures were available. Use of outcome measures as identified through the audit did not change. CONCLUSIONS: The "All About Outcomes" software is clinically useful. Further research should evaluate whether using the software affects the use of outcome measures in clinical practice. PMID- 10378486 TI - Constraint-induced movement therapy for motor recovery in chronic stroke patients. AB - OBJECTIVE: Assessment of the effectiveness of constraint-induced (CI) movement therapy and quantitative evaluation of the effects of CI therapy. DESIGN: Intervention study; case series; pretreatment to posttreatment measures and follow-up 3 months after intervention. SETTING: An outpatient department. PATIENTS: Five chronic stroke patients with moderate motor deficit; convenience sample. INTERVENTIONS: CI therapy consisting of restraint of the unaffected upper extremity in a sling for 14 days combined with 6 hours of training per weekday of the affected upper extremity. MAIN OUTCOME MEASURES: Actual Amount of Use Test (AAUT), Motor Activity Log (MAL), Wolf Motor Function Test (WMFT), and Arm Motor Ability Test (AMAT) RESULTS: There was a substantial improvement in the performance times of the laboratory tests (AMAT, WMFT, p < or = .039) and in the quality of movement (AMAT, WMFT, p < or = .049; MAL, p = .049), particularly in the use of the extremity in "real world" environments (AAUT, p = .020), supported by results of quantitative evaluation. The effect sizes were large and comparable to those found in previous studies of CI therapy. CONCLUSIONS: CI therapy is an efficacious treatment for chronic stroke patients, especially in terms of real world outcome. PMID- 10378488 TI - Correlation of clinical and ultrasonographic features in congenital muscular torticollis. AB - OBJECTIVE: To find the relationship between the ultrasonographic pictures and the clinical features of patients with congenital muscular torticollis (CMT). DESIGN: Prospective survey of patients with clinically suspected CMT by high-resolution ultrasonography. SETTING: Rehabilitation department of a tertiary care center. PARTICIPANTS: Two hundred fifty-six CMT patients, from the ages of 9 days to 16yrs, with a mean follow-up period of 6.7 months. MAIN OUTCOME MEASURES: Correlation of the ultrasound appearance of the involved sternocleidomastoid (SCM) muscles with clinical features. The pathologic findings in diseased muscles from patients who underwent surgical intervention were also evaluated. RESULTS: Muscle abnormalities were identified ultrasonographically in 218 CMT patients (85%) and were classified into four types: a fibrotic mass in the involved muscle (type I, 15%); diffuse fibrosis mixing with normal muscle (type II, 77%) and without normal muscle in the involved muscle (type III, 5%); and a fibrotic cord in the involved muscle (type IV, 3%). Compared with type I patients, type IV patients were more likely to undergo surgical treatment (odds ratio = 31.54, p = .0196). Type III patients were more likely to undergo surgical treatment, although this tendency was not statistically significant. CONCLUSION: Ultrasonography can precisely identify pathologic changes in the involved SCM muscle of CMT patients. Type III and IV patients are more likely to require surgical intervention. PMID- 10378489 TI - Do Medicare patients with disabilities receive preventive services? A population based study. AB - OBJECTIVE: To compare health maintenance procedure rates of Medicare patients with different levels of disability. STUDY DESIGN: Observational study analyzing data from the 1995 Medicare Current Beneficiary Survey (MCBS, n = 15,590). Self reported Pap smears, mammograms, and influenza and pneumococcal vaccinations were compared between groups with different levels of health-related difficulties in six activities of daily living (ADL). RESULTS: Compared to those without disabilities, the most severely disabled women (limitations in 5 or 6 ADL) reported fewer Pap smears (age < or =70, 23% vs 41%, p < .001) and mammograms (age > or = 50, 13% vs 44%, p < .001). In a controlled analysis, individuals with this high level of disability were 57% (95% confidence interval [CI], 33% to 72%) and 56% (95% CI, 43% to 76%) less likely to report receiving Pap smears and mammograms, respectively, compared with able-bodied women, regardless of their age, whether they were in an HMO, or whether they lived in a long-term care facility. Functional limitations were not a deterrent to receiving vaccinations. In general, patients in HMOs reported more procedures than those in fee-for service, while those in long-term care facilities reported fewer procedures than those living in the community. CONCLUSIONS: Disability among Medicare patients is a significant, independent risk factor for not receiving mammograms and Pap smears. Efforts should be made to identify the most severely disabled because they are at particular risk. PMID- 10378490 TI - Laser therapy: a randomized, controlled trial of the effects of low-intensity Nd:YAG laser irradiation on musculoskeletal back pain. AB - OBJECTIVE: To assess the effectiveness of low-intensity laser therapy in the treatment of musculoskeletal low back pain. DESIGN: A double-masked, placebo controlled, randomized clinical trial. SETTING: A physical medicine and rehabilitation clinic. PARTICIPANTS: Sixty-three ambulatory men and women between the ages of 18 and 70yrs with symptomatic nonradiating low back pain of more than 30 days' duration and normal neurologic examination results. INTERVENTION: Subjects were bloc randomized into two groups with a computer-generated schedule. All underwent irradiation for 90 seconds at eight symmetric points along the lumbosacral spine three times a week for 4 weeks by a masked therapist. The sole difference between the groups was that the probes of a 1.06 microm neodymium:yttrium-aluminum-garnet laser emitted 542mW/cm2 for the treated subjects and were inactive for the control subjects. MAIN OUTCOME MEASURES: Subject's perception of benefit, level of function as assessed by the Oswestry Disability Questionnaire, and lumbar mobility. RESULTS: The treated group had a time-dependent improvement in two of the three outcome measures: perception of benefit and level of function. These results were most marked at the midpoint evaluation (p < .005, p < .01) and end of treatment (p < .017, p < .001) but tended to lessen at the 1-month follow-up (p < .10, p < .004). Lumbar mobility did not differ between the groups at any time. All tests were two-sample t tests with unequal variances. CONCLUSIONS: Treatment with low-intensity 1.06 microm laser irradiation produced a moderate reduction in pain and improvement in function in patients with musculoskeletal low back pain. Benefits, however, were limited and decreased with time. Further research is warranted. PMID- 10378491 TI - Assessment of command-following in minimally conscious brain injured patients. AB - OBJECTIVE: To develop a method for establishing the presence of command-following in individuals with traumatic brain injury, based on the principles of single subject experimental design. DESIGN: A series of single-subject experiments, individualized to the particular command-following question about a particular patient. SETTING: An inpatient rehabilitation hospital with a specialized program for vegetative and minimally conscious brain injured patients. PATIENTS: Eight individuals with serious brain injury of traumatic or nontraumatic origin, presenting in vegetative or minimally conscious states. INTERVENTIONS: The frequency of performance of the behavior in question was assessed in response to commands and in relation to appropriate control conditions. Data were analyzed with chi2 or Fisher's exact test, as well as measures derived from signal detection theory. MAIN OUTCOME MEASURES: The frequency of performance of a specific behavior in the presence of a command and in relevant contrasting conditions. RESULTS: This method identified whether a specific behavior was being performed in response to command and whether the reliability of this behavior was changing over time either spontaneously or in response to treatment. CONCLUSIONS: Quantitative assessment of command-following based on principles of single subject experimental design can determine whether patients are capable of following commands and whether this ability changes over time or in response to treatment. PMID- 10378492 TI - Improving cognitive function after brain injury: the use of exercise and virtual reality. AB - OBJECTIVE: To assess the impact of exercise and virtual reality (VR) on the cognitive rehabilitation of persons with traumatic brain injury (TBI). DESIGN: Before-after trial assessed cognitive function after a 4-week intervention program. A random allocation crossover assessed changes in reaction and movement times after a single bout of VR exercise and a no-exercise control condition. SETTING: Brain injury rehabilitation unit in Edinburgh, Scotland. PATIENTS: (1) Four-week intervention: a consecutive sample of 13 suitable TBI adults were compared to control populations (n > 25) of previous TBI patients of similar age, severity, and time postinjury. (2) Single-bout intervention: a consecutive sample of 13 suitable adults with moderate TBI, 6.29 to 202.86 weeks postinjury. INTERVENTION: Nonimmersive VR exercise. MAIN OUTCOME MEASURES: (1) Tests of attention, information processing, learning, and memory. (2) Reaction and movement times. RESULTS: After the 4-week intervention patients performed significantly better than controls on the digit symbol (p < .01). verbal (p < .01), and visual learning tasks (p < .05). Significant improvements in reaction times (p < .01) and movement times (p < .05) were gained following a single bout of VR exercise. CONCLUSION: Exercising in a virtual environment offers the potential for significant gains in cognitive function. PMID- 10378493 TI - Practice effects on the less-affected upper extremity after stroke. AB - OBJECTIVE: To test the hypotheses that (1) adults who have had a stroke, using the less affected upper extremity (UE), improve performance of an aiming task with practice, and (2) compared with control subjects, stroke patients show less improvement in a complex condition. DESIGN: Movement time (MT) and kinematic data were collected over practice. Comparisons were made between the less-affected UE of stroke patients and the same hand of controls. SETTING: A human performance laboratory. PARTICIPANTS: A matched sample of right-handed adults, 10 with unilateral stroke and 10 nondisabled controls. INTERVENTION: Practice of an aiming task in an easy and complex condition as defined by target width and distance between two targets. MAIN OUTCOME MEASURES: MT, peak velocity, and temporal phases of the trajectory. RESULTS: Adults who had experienced a stroke had persistently longer MTs than control subjects; however, all participants achieved faster MTs with practice in both conditions. The absolute amount of time in each temporal phase decreased without a change in the relative times. Peak velocity increased only in the easy condition. CONCLUSIONS: Adults with stroke damage can improve motor performance of the less-affected UE with practice. Further study is needed to see if practice effects are permanent and generalizable. PMID- 10378494 TI - Measurement of mood states in stroke patients: validation of the visual analog mood scales. AB - OBJECTIVE: The Visual Analog Mood Scales (VAMS) were recently developed by Stern and colleagues to assess mood state in neurologically impaired patients. These brief scales require that a patient place a single pen mark along a 100mm vertical line to indicate how he or she presently feels. Although previous studies have garnered evidence in support of the validity of these brief scales when administered to psychiatric patients and healthy young adult and geriatric control subjects, it is presently unknown whether the VAMS are valid measures of internal mood state in neurologically impaired stroke patients. The purpose of the present investigation was to assess reliability and validity of the VAMS in a stroke-patient population. PARTICIPANTS: Participants were 41 (21 men and 20 women) inpatients admitted for either acute stroke or rehabilitation following stroke. DESIGN: Participants completed both the VAMS and a modified version of the Profile of Mood States (POMS). Nonparametric multitrait-multimethod analyses were performed using the Pearson correlations among and between the six subscales of the VAMS and the POMS. CONCLUSION: The VAMS possess good convergent and discriminant validity when administered to stroke inpatients, providing further support for the utility of these brief, easily administered scales. PMID- 10378495 TI - The predictive value of plantar flexion of the toes in the assessment of neuropathic voiding disorders in patients with spine lesions at the thoracolumbar level. AB - OBJECTIVE: To correlate the presence of voluntary contraction of the plantar flexors of the toes with neuropathic voiding disorders in patients with spinal cord injury (SCI) after thoracolumbar fracture. SUBJECTS AND METHODS: Sixty-three SCI patients with thoracolumbar fractures were prospectively examined neurologically (American Spinal Cord Injury Association protocol) and urodynamically during their first hospitalization. To assess neurologic recovery, patients were reassessed after at least 1 year. Bladder function was also reevaluated and correlated to neurologic evaluation performed on the same day. SETTINGS: Primary care center, university facility. RESULTS: There was a significant correlation between score of the plantar flexors of the toes and the presence or absence of voluntary contraction of the external anal/urethral sphincter (p < .001). However, this was of no predictive value concerning neuropathic bladder type (p > .05). Seven patients recovered from their neuropathic voiding disorders. There was a significant correlation between the reappearance of a voluntary external anal/urethral sphincter contraction and bladder recovery (p < .01), but there was no significant correlation between the score obtained for the plantar flexors of the toes and the presence or absence of the bulbocavenosus reflex (p > .05). CONCLUSIONS: It appears that in SCI patients with thoracolumbar fractures, the presence of a voluntary contraction of the plantar flexors of the toes correlates with active contraction of the external anal/urethral sphincter but does not enable differentiation of the types of neuropathic voiding disorders. Urodynamic examination remains mandatory for assessment of the functional aspect of the neuropathic voiding disorders. PMID- 10378496 TI - Late onset polio sequelae: disabilities and handicaps in a population-based cohort of the 1956 poliomyelitis outbreak in The Netherlands. AB - OBJECTIVE: To investigate the prevalence of new neuromuscular symptoms, disabilities, and handicaps in a group of polio survivors. DESIGN: A self constructed health questionnaire about neuromuscular complaints and disability and handicap levels during the stable period after recovery from polio and at present. SUBJECTS: Three hundred fifty subjects, derived from the 1,784 polio cases registered during the 1956 polio outbreak in The Netherlands. RESULTS: Respondents totaled 260 (74%), 27 of whom denied or did not recall having had paralytic poliomyelitis. The remaining 233 subjects comprised the study group (mean age, 44yrs; range, 39 to 77; SD = 6.3). Frequency of all neuromuscular complaints at present time was significantly higher than that during the stable period after polio (range in p of .001 to .004). Fifty-eight percent of cases reported an increase in muscle weakness in comparison with muscle condition during the stable period. Fifty-six percent reported an increase in disabilities, mainly a restriction in gait functions. Fifty-three percent reported increased handicaps with regard to occupation and social integration, and there was an increased need for adaptive measures and devices. CONCLUSION: Nearly 60% of a sample of Dutch survivors of the 1956 polio outbreak experience late onset polio sequelae, resulting in increased severity of disabilities and handicaps. PMID- 10378497 TI - Exercise intensity: its effect on the high-density lipoprotein profile. AB - OBJECTIVE: To determine the effect of aerobic exercise intensity on the active subfraction of serum high-density lipoprotein (HDL) concentration. DESIGN: A randomized control, before-and-after investigation that tested the hypothesis that high-intensity exercise training would result in improvements in serum concentrations of HDL subfraction 2 (HDL2) greater than those accompanying moderate-intensity training. SETTING: Exercise tests were completed in a hospital stress testing laboratory, and cholesterol analyses were performed in a university research laboratory. Exercise training was performed in the community at a site determined by the subject. SUBJECTS: Subjects were 25 healthy female employees of a teaching hospital. INTERVENTION: Maximum treadmill tests and serum cholesterol profiles were assessed in 25 women before and after a 12-week aerobic walking regimen; 12 women in a high-intensity exercise group (HIG) walked at a target heart rate of 80% and 13 women in a moderate-intensity exercise group (MIG) walked at a heart rate of 60% of their heart rate reserve for a distance of 2 miles three times weekly. MAIN OUTCOME MEASURES: The main dependent variable was HDL2; other measures of the HDL profile were total HDL and HDL3. Peak oxygen uptake (VO2) was also evaluated as a dependent variable to ensure a general aerobic adaptation resulted from the exercise regimen. Measures were analyzed as pretraining to posttraining change scores and absolute values using independent and dependent t tests as appropriate. Statistical significance was assigned atp < .05. RESULTS: Total HDL was 32.3+/-8.5mg/dL before and 40.3+/-10.6mg/dL after training in the MIG and 31.6+/-6.2mg/dL before and 38.2+/-12.0mg/dL after training in the HIG. HDL2 was 14.2+/-5.7mg/dL before and 18.5+/-6.9mg/dL after training in MIG. HDL2 was 13.0+/-6.2mg/dL before and 19.6+/-8.9mg/dL after training in the HIG. Total HDL and HDL2 increased significantly in both groups as a result of exercise training, and intragroup differences were not observed. HDL3 was not affected by exercise training. Training resulted in significant increases in peak VO2 in both MIG and HIG (29.0+/-5.0 to 31.9+/-5.4mL/kg/min in the MIG and 30.7+/-5.2 to 33.5+/-6.3mL/kg/min in the HIG). Intergroup differences in change scores for peak VO2, HDL, and HDL2 were not observed. CONCLUSION: The results and analyses did not support the hypothesis that the HIG would acquire increases in HDL2 profile beyond those observed for the MIG. Moderate-intensity training was sufficient to improve the HDL profile, and high-intensity training appeared to be of no further advantage as long as total training volume (total walking distance per week) was constant. PMID- 10378498 TI - Gait consistency over a 7-day interval in people with Parkinson's disease. AB - OBJECTIVE: To evaluate the consistency of temporal and spatial parameters of the walking pattern in subjects with idiopathic Parkinson's disease (PD) over a 7-day interval during the "on" phase of the levodopa medication cycle. SETTING: Walking patterns were measured on a 12-meter walkway at the Kingston Gait Laboratory, Cheltenham, using a computerized stride analyzer. SUBJECTS: Sixteen subjects (7 women, 9 men) with PD recruited from the Movement Disorders Clinic at Kingston Centre. MAIN OUTCOME MEASURES: Speed of walking, stride length, cadence, and the percentage of the walking cycle spent in the double limb support phase of gait were measured, together with the level of disability as indexed by the modified Webster scale. RESULTS AND CONCLUSIONS: Product-moment correlation coefficients and intraclass correlation coefficients (ICC 2,1) for repeat measures over a 7 day interval were high for speed (r = .90; ICC = .93), cadence (r = .90; ICC = .86), and stride length (r = 1.00; ICC = .97) and moderate for double limb support duration after removal of outliers (r = .75; ICC = .73); 95% confidence intervals for the change scores were within clinically acceptable limits for all variables. The mean modified Webster score was 11.4 on the first day and 10.1 7 days later. The gait pattern and level of disability in subjects with PD without severe motor fluctuations remained stable over a 1-week period when optimal medication prevailed. PMID- 10378499 TI - Changes of laboratory markers of cognitive brain function by thermostimuli in the elderly. AB - OBJECTIVE: Plain external applications of physical stimuli, which are used quite commonly in geriatric care in Germany, have not been studied for their influence on cognitive brain function. The aim of this randomized crossover study was to examine the influence of dermatoreceptive stimuli on cognitive brain function in healthy geriatric volunteers. METHODS: Twenty-four healthy volunteers (23 women, 1 man) were randomized into two groups (crossover design). Group A (mean age, 68.8+/-6.2 [SD] years) was treated with a 10 degrees C to 12 degrees C cold stimulus for 10 seconds (a so-called "Kneipp face shower"), followed by a cold 10 degrees C to 12 degrees C wetpack at the neck for 1 minute. Group B (mean age, 69.8+/-5.3 [SD] years) was subjected to an identical procedure but with warm to neutral temperatures of 34 degrees C to 36 degrees C. After I week the two groups were interchanged. The parameters of interest were the critical flicker frequency (CFF) and the latencies of the event-related P-300 potentials of the visual evoked potentials (VEP), which can be considered an electroencephalographic marker of the cognitive functional ability. The CFFs and the P-300 latencies and amplitudes were measured directly both before and 10 minutes after the application of the respective stimuli. In addition, the CFFs were recorded 30 and 60 minutes later. RESULTS: After cold water stimuli were applied, the CFF increased from 32.55+/-2.26/sec (mean+/-SD) to 33.06+/-2.25/sec (p = .003) 10 minutes after the stimulus. Thirty minutes later the CFF was still elevated at 32.95+/-2.3/sec (p = .043). The P-300 latencies, after cold water application, decreased by 4.8% (p < .001), from 266.5+/-21.1msec (mean+/-SD) to 253.7+/ 16.9msec. After warm stimuli they increased from 258.69+/-14.8msec to 266.17+/ 20.1msec (p = .01). The P-300 amplitudes were significantly elevated, by 5% (p = .004), only after cold stimuli. CONCLUSION: Cold water applied locally to the face and neck region can provoke significant changes in electroencephalographic markers as measured by an electroencephalographic marker (VEP and P-300 latency) and, by inference, may help to improve cognitive function in the elderly. PMID- 10378500 TI - Enabling factors related to prosthetic use by people with transtibial and transfemoral amputation. AB - OBJECTIVE: To evaluate the frequency and extent of prosthetic use by people with lower limb amputation and identify factors that facilitate prosthetic use. DESIGN AND SETTING: Five-year follow-up survey using the Prosthetic Profile of the Amputee (PPA) questionnaire and Dillman's mailing strategy. SUBJECTS: Adults with unilateral transtibial and transfemoral amputation (n = 396) who had completed a prosthetic training program. MAIN OUTCOME MEASURES: Frequency of prosthetic wear, in hours per week, and active prosthetic use for locomotion indoors and outdoors. RESULTS: Eighty-five percent of the respondents (mean age 62.9+/-15.9yrs) were prosthetic wearers; 53% used their prosthesis for locomotion indoors, and 64% outdoors. Ability to don the prosthesis (p < .001), locomotor capabilities with the prosthesis (p < .001), walking distances (p < .001), automaticity of gait (p < .05), and assistive devices used (p < .001) were the main factors related to the three outcome measures. People with transfemoral amputation reported greater difficulties in donning their prosthesis (p < .01) and a significantly higher rate of falls (p < .001). CONCLUSION: The majority of people with lower limb amputation wear their prosthesis daily. With the exception of resources (prosthetic laboratory and means of transportation), all enabling factors investigated were significantly associated with the outcome measures. PMID- 10378501 TI - Inhibitory casting decreases a vibratory inhibition index of the H-reflex in the spastic upper limb. AB - OBJECTIVE: To test the hypothesis that application of an inhibitory cast to the spastic upper limb will decrease a vibratory inhibition index (VII) of the H reflex in the spastic upper limb. DESIGN: Prospective, nonrandomized, open-label trial. SETTING: University tertiary care center. PARTICIPANTS: Eight adults with upper limb spasticity. INTERVENTION: Fiberglass cast application spanning the wrist to the upper arm. MAIN OUTCOME MEASURE: The amplitude of the H-reflex with and without continuous 60Hz vibration to the tendon of the flexor carpi radialis was measured, and the VII was calculated using the formula: [H-reflex amplitude (vibrated)/H-reflex amplitude (control)] x 100%. RESULTS: Mean VII decreased from baseline (70.7) on day 1 (67.6, p = .699), day 2 (55.9, p =.066), and day 3 (43.5, p = .033) of casting, and increased on day 4 (89.9, p = .146) after removal of the cast. CONCLUSION: Findings lend support to the idea that during application of an inhibitory cast motor neuron excitability is decreased in the spastic upper limb. PMID- 10378502 TI - Unilateral lower limb injury: its long-term effects on quadriceps, hamstring, and plantarflexor muscle strength. AB - OBJECTIVE: To ascertain if long-term deficits in quadriceps, hamstring, and plantarflexor muscle strength remain after unilateral lower-limb musculoskeletal injury and to quantify whether improvements in performance continue once a subject concludes rehabilitation and returns to everyday activities. The relation between the size of decrement and limb dominance, type of injury, and time since injury was also considered. DESIGN: Isometric and/or dynamic muscle strength of both legs was measured (using the KinCom 500H isokinetic dynamometer) in 48 subjects. SETTING: A physiological laboratory at Brunel University. PATIENTS: Patients were recruited locally via a district general hospital, sports injury clinic, and university. MAIN OUTCOME MEASURES: Muscle strength in the injured limb, reported as a percentage of muscle strength in the uninjured limb. It was assumed that the preinjury state of the injured limb was similar to that of the uninjured limb. RESULTS: Decrements were seen in mean isometric and peak isometric, concentric, and eccentric quadriceps activity (p < .0001) and isometric plantarflexor activity (p < .05) in the injured limb, with the type of injury influencing the size of the decrement. Minimal difference was found in the hamstring muscles. CONCLUSIONS: The decrements in performance in the quadriceps muscle imply that full recovery (as defined by the preinjury state) is frequently not achieved and stress the need for accurate, objective assessment of muscle strength and further investigation into the nature and duration of rehabilitation after musculoskeletal injury. PMID- 10378503 TI - An unusual extraspinal cause of bilateral leg pain. AB - Low back pain with pain radiating to the lower extremities is common in patients referred to a spine center. Lumbar spine pathology is commonly the etiology of such symptoms, but extraspinal causes of back and leg pain can manifest as a radicular disorder. Extraspinal etiologies must be considered in the workup of back and leg pain. This report describes an unusual case of spontaneously occurring bilateral femoral neck stress fractures presenting as low back pain with seemingly bilateral L4 radicular symptoms. PMID- 10378504 TI - Walker inlet-closure strap for unsteady patients with lower-extremity amputations. AB - Some patients with lower-extremity amputations who use a walker fall backwards after advancing too far forward into the walker's base of support. In a pilot study of 14 patients with unilateral lower-extremity amputations who stepped into the forward two thirds of the walker base, this problem was corrected by using a knee-high elastic strap to close the open posterior "inlet" of the walker. Without the strap, the stance-phase position of the leading ankle was in the forward third of the walker base for 8 subjects and in the middle third for 6. With the walker strap, the ankle position was in the middle third for one subject, in the posterior third for 6, and at or behind the walker inlet for 7 (p < .0002). This simple intervention appears to correct the potentially dangerous behavior of stepping too far into the walker base. PMID- 10378505 TI - Misfolded proteins in the endoplasmic reticulum. PMID- 10378506 TI - Identification of a metaplastic cell lineage associated with human gastric adenocarcinoma. AB - Metaplastic cell lineages arising in response to chronic injury are precursors for the evolution of dysplasia and adenocarcinoma. Although a subtype of intestinal metaplasia has been associated with gastric adenocarcinoma, the link between this lineage and the evolution of gastric adenocarcinoma has remained unclear. Wang et al (1998) have reported that an aberrant metaplastic cell lineage with morphological characteristics similar to Brunner's glands of the duodenum develops in the fundic mucosa of mice infected with Helicobacter felis. This metaplastic lineage expresses the trefoil peptide spasmolytic polypeptide (SP). Given the epidemiological association of Helicobacter species infection with gastric cancer, we hypothesized that this SP-expressing metaplastic (SPEM) lineage may represent a precursor to or appear commensurate with gastric adenocarcinoma. The SPEM lineage was present in 68% of fundic biopsies from patients with fundic Helicobacterpylori-associated gastritis, but was absent in biopsies of fundic mucosa from patients without H. pylori infection. In a review of archival samples from 22 resected gastric adenocarcinomas, we found the SPEM lineage in 91% of cases, typically located in mucosa adjacent to the carcinoma or areas of dysplasia. Importantly, 59% of resections showed SP immunoreactivity within dysplastic cells. These data indicate a strong association of the SPEM lineage with both chronic H. pylori infection and gastric adenocarcinoma. PMID- 10378507 TI - Expression of the Wnt gene family during late nephrogenesis and complete ureteral obstruction. AB - Because the Wnt-4, -7b, and -11 genes are expressed in metanephric kidneys and code for secreted glycoproteins that may serve as mediators of the transformation of renal mesenchyme to epithelium, we investigated the pattern of Wnt gene expression in late metanephrogenesis and after ureteral obstruction. Newborn and 10-, 20-, and 60-day-old rats underwent complete unilateral ureteral ligation or sham operation. The kidneys were collected bilaterally 1, 5, 10, 20, or 30 days later. RNase protection assays were used to quantify the amounts of mRNA encoding Wnt-4, -7b, and -11, E-cadherin, and cytokeratin-19. Renal development was assessed by histologic characterization of vimentin, cytokeratin, E-cadherin, and beta-catenin distribution. During normal development, the amounts of mRNA encoding Wnt-4 and Wnt-11 increased during gestation and then abruptly decreased after the completion of metanephrogenesis, 15 days after birth. In contrast, the amounts of mRNA encoding Wnt-7b, E-cadherin, and cytokeratin-19 increased during development and into adulthood. In neonatally obstructed kidneys, the expression of Wnt-4 was abnormally maintained when obstruction was induced before the completion of renal development and was reactivated when obstruction was induced after the completion of metanephrogenesis. Wnt-7b expression was minimally affected and Wnt-11 expression was only transiently affected by obstruction. In neonatally obstructed kidneys, the differentiation of mesenchyme to epithelium failed to proceed normally, with the majority of cells maintaining vimentin expression and some differentiated epithelial cells reverting to vimentin expression. In addition, the expression of E-cadherin and cytokeratin was increased in epithelial cells. Changes in the expression of Wnt genes were correlated with histologic changes. This study suggests that Wnt-4 and -11 are likely to be important mediators of the transformation of mesenchyme to epithelium in the kidney. Obstruction induced during metanephrogenesis disrupts the normal pattern of Wnt-4, -7b, and -11 expression and interferes with the normal transformation process in developing kidneys, by maintaining the mesenchymal component and inducing the transformation of epithelium to mesenchyme. PMID- 10378508 TI - Endothelial expression of endothelial nitric oxide synthase and endothelin-1 in human coronary artery disease. Specific reference to underlying lesion. AB - Nitric oxide and endothelin-1 (ET-1) are two major endothelium-derived factors with opposing effects on the function and structure of the vessel wall. We investigated the endothelial expression of endothelial nitric oxide synthase (eNOS) and ET-1 in coronary artery disease (CAD) with special reference to the types of underlying lesions. Immunohistochemistry and in situ hybridization were performed in coronary arteries of heart transplant recipients with (n = 16) and without (n = 11) CAD. All coronary arteries from patients with CAD (n = 23) had concentric fibrous or advanced lesions, whereas most of the arteries (25 of 31) from patients with non-CAD showed normal appearance (myointimal thickening only) or eccentric lesions alone. Normal coronary segments consistently showed apparent endothelial immunoreactivity and mRNA signals for both eNOS and ET-1. In atherosclerotic coronary segments, endothelial expression of eNOS and ET-1 was reduced in most lesion sites, particularly in severe subendothelial lesions with dense fibrosis or macrophage accumulation, but not with smooth muscle cells only. Conversely apparent ET-1, compared with weak or focal eNOS signals, were more frequently seen in coronary segments with concentric severe lesions from CAD but not non-CAD patients. Immunoreactivity and mRNA signals for ET-1 were co localized with those for ET converting enzyme-1 in the endothelium, as well as in the underlying macrophages and smooth muscle cells. These results indicate the presence of differential endothelial expression of eNOS and ET-1 in diseased human coronary arteries with severe concentric atherosclerotic lesions, a finding that was rare in atherosclerotic lesions of coronary arteries of non-CAD patients. Altered expression of endothelium-derived factors may contribute to abnormality of coronary vasomotor tone and the formation of subendothelial lesions in CAD. PMID- 10378509 TI - Loss of heterozygosity in 11q13-14 regions in gastric neuroendocrine tumors not associated with multiple endocrine neoplasia type 1 syndrome. AB - Loss of heterozygosity (LOH) at the MEN1 gene locus at 11q13 is commonly found in type II gastric carcinoid tumors, which are associated with multiple endocrine neoplasia type 1 (MEN-1). In contrast, information is scanty or absent for other types of gastric neuroendocrine tumors, represented by type I carcinoids (associated with chronic atrophic gastritis), type III (sporadic) carcinoids, and neuroendocrine carcinomas. Moreover, LOH analysis of the allelic region distal to the MEN1 gene, which is postulated to contain an additional tumor suppressor gene effective in MEN-1-associated and sporadic endocrine tumors, has never been performed. To clarify these issues, DNA extracted from archival tissue from 25 type I carcinoids, 4 type III carcinoids, and 2 neuroendocrine carcinomas was amplified by PCR, using primers for six polymorphic markers located on chromosome 11q13 (PYGM, D11S4946, and D11S913) and 11q14 (D11S916, D11S901, and D11S1365), for analysis of LOH. Allelic losses in the 11q13-14 region with at least two polymorphic markers were found in 12 of 25 (48%) type I carcinoids. When LOH was found in the 11q13 region, it was large and continuous and extended to the most telomeric marker investigated. In one tumor, retention of heterozygosity for markers in the MEN1 region and LOH for distal markers were observed. No LOH was found in three of four type III carcinoids. Large deletions in both the 11q13 and 11q14 regions were observed in both neuroendocrine carcinomas investigated. In conclusion, LOH in the 11q13-14 regions is frequently found in type I carcinoids and neuroendocrine carcinomas of the stomach, suggesting the involvement of the MEN1 gene and/or a more telomeric tumor suppressor gene in the pathogenesis of these non-MEN-1-associated neuroendocrine tumors. The low rate of LOH at 11q13-14 suggests the predominance of different genetic mechanisms in type III carcinoids, which also differ from other types of gastric carcinoids in the lack of a promoter role for gastrin. PMID- 10378510 TI - Thrombopoietin expression in normal and hypobaric hypoxia-induced thrombocytopenic rats. AB - Thrombopoietin (TPO) is important as the physiologic regulator of platelet production. High-altitude hypoxia is a well-known cause of polycythemia and thrombocytopenia in animals. Fifty-two Wistar rats were housed for 0.5 to 21 days in a mechanical chamber in an environment equivalent to that found at 5500 m to determine (a) the cellular localization of TPO and (b) whether the decreased platelet and megakaryocyte counts in rats exposed to a hypobaric hypoxic environment (HHE) are associated with an altered TPO mRNA expression. In normal rats, there were high levels of TPO mRNA in the liver and kidney, intermediate levels in the brain and large intestine, and low levels in the skeletal muscle and small intestine. TPO mRNA and protein were expressed in Purkinje cells and neuronal cells in the brain, in proximal tubular cells and the mesangial cells of the glomeruli in the kidney, in hepatocytes and biliary duct epithelial cells, in absorptive epithelial cells in the large intestine, in the epidermis, and in the lung. The platelet count in the blood and megakaryocyte counts in the bone marrow and spleen were all decreased significantly after 5 or more days of exposure to HHE. In major producers such as the liver and kidney and in minor producers such as the brain, TPO mRNA levels, which tended to be decreased after 0.5 to 3 days of exposure to HHE, had returned to normal by about Day 5 or 7. Thus, during the HHE period with a decreased platelet count, no changes in TPO mRNA levels were detected in these three organs. In conclusion, we have demonstrated that TPO production occurs in various types of cells. In HHE, however, factors other than TPO may be involved in hypobaric hypoxia-induced thrombocytopenia in rats. PMID- 10378511 TI - A mouse prion protein transgene rescues mice deficient for the prion protein gene from purkinje cell degeneration and demyelination. AB - Disruption of both alleles of the prion protein gene, Prnp, renders mice resistant to prions; in a Prnp o/o line reported by some of us, mice progressively developed ataxia and Purkinje cell loss. Here we report torpedo like axonal swellings associated with residual Purkinje cells in Prnp o/o mice, and we demonstrate abnormal myelination in the spinal cord and peripheral nerves in mice from two independently established Prnp o/o lines. Mice were successfully rescued from both demyelination and Purkinje cell degeneration by introduction of a transgene encoding wild-type mouse cellular prion protein. These findings suggest that cellular prion protein expression may be necessary to maintain the integrity of the nervous system. PMID- 10378512 TI - Identification of ataxia telangiectasia heterozygotes, a cancer-prone population, using the single-cell gel electrophoresis (Comet) assay. AB - Heterozygotes of ataxia telangiectasia (AT) may comprise up to 1% of the general population. Because these individuals have no clinical expression of AT but may be highly radiosensitive and strongly predisposed for several forms of cancer, identification of AT carriers represents a considerable interest in cancer epidemiology and radiotherapy. We report a new approach for the in vitro identification of AT-heterozygotes based on the evaluation of the radiosensitivity and DNA damage repair ability of peripheral blood mononuclear cells using the single-cell gel electrophoresis (Comet) assay. The assay was performed on cells isolated from four different groups of individuals: (1) apparently healthy donors (n = 10); (2) patients with breast cancer showing a normal reaction to radiotherapy (n = 10); (3) a group of obligate AT carriers (parents of AT-homozygotes, n = 20); and (4) AT-homozygotes (n = 4). Cells irradiated with 3 Gy of x-rays were assayed for three parameters: (1) the initial and (2) residual DNA damage and (3) the kinetics of DNA damage repair. Both AT heterozygotes' and AT-homozygotes' cells were found to be highly sensitive to x irradiation. Quantitative evaluation of the single-cell electrophoregrams revealed that the average initial DNA damage in AT-heterozygous and AT-homozygous cells was almost three times higher than that in control non-AT cells. In addition, the DNA repair process in irradiated AT carrier cells was almost three times slower, and the extent of irreparable DNA damage in these cells was three times greater than in controls. Simultaneous assessment of the three parameters enabled correct identification of all tested AT carriers. This method seems to be a sensitive and useful tool for populational studies as a rapid prescreening test for a mutated AT status. The approach can also be extended for prediction of the in vivo radiosensitivity, which would enable optimization of individual radiotherapy schedules. PMID- 10378513 TI - Semi-quantitative fluorescence in situ hybridization analysis indicates that the myc protein is consistently stabilized both before and after transformation of low-grade follicular center to high-grade diffuse large cell lymphoma. AB - We have investigated the expression of the MYC gene at both the mRNA and protein levels to determine how these parameters are related in lymphoma cells and in nonmalignant lymphoid cells. To do this we have adopted a multicolor fluorescence in situ hybridization methodology, which has allowed us to investigate the expression of different genes at the same time in the same cell. We have made use of the digital imaging capabilities of a charge-coupled device camera system to quantify the hybridization signals for the MYC gene and, by comparing these to the expression of a control gene (glyceraldehyde-3-phosphate dehydrogenase; GAPDH), have obtained relative quantitations of MYC mRNA and protein levels. In this study we have compared cells both within and outside the germinal centers in control tissues (reactive lymph nodes and tonsils) and in low-grade follicular center lymphomas, as well as cells in high-grade diffuse large cell lymphomas. The MYC/GAPDH mRNA hybridization signal ratios were calculated and found to be higher in cell populations containing a majority of malignant cells (p < 0.04). However, when the myc/GAPDH protein hybridization signal ratios were calculated, these were significantly higher in malignant cells from all lymphomas than the ratios observed in the nonmalignant cells (p < 0.0005). These observations indicate that the environment in a malignant cell may contribute to the stabilization of the myc protein, thus enabling it to function for a longer time period than in nonmalignant cells. PMID- 10378514 TI - Presence of urokinase-type plasminogen activator receptor in urine of cancer patients and its possible clinical relevance. AB - High levels of urokinase-type plasminogen activator receptor (uPAR) are expressed in various types of cancer. Recent studies showed that cancer patients may have increased levels of soluble (s)uPAR in their serum. In the present study, we show that urine samples from healthy volunteers contain measurable amounts of suPAR. suPAR/creatinine levels from healthy controls showed only little variation over the day and were even stable during a month of continued monitoring. Importantly, urinary suPAR/creatinine levels were highly correlated with serum suPAR concentrations. Urinary suPAR levels were elevated in patients with different types of cancer. Interestingly, part of the urinary suPAR seemed to be present in a cleaved form, as has been found in tumor tissue extracts. Together with the recently established, cell migration-promoting effect of certain cleaved fragments of suPAR, the present data suggest that the measurement of urinary suPAR and/or its cleaved forms might have clinical implications. PMID- 10378515 TI - Characteristics of nonmalignant and malignant human prostate in organ culture. AB - Prostate tissue was obtained from 52 radical prostatectomies immediately upon surgery. From each specimen, a small piece of tissue was fixed in 10% buffered formalin and used for histology, cytokeratin staining, staining with the antibodies to the proliferation-associated antigen (Ki-67), and histochemical evaluation of the epithelial-stromal basement membrane. A second piece was used for the isolation of epithelial cells and stromal cells in monolayer culture. The remainder of each specimen was cut into cubes (approximately 1 mm on a side) and incubated in organ culture for up to 20 days. At the end of the incubation period, tissue was fixed in 10% buffered formalin and examined as described above with zero-time tissue. These studies showed that normal epithelial and stromal elements survived in organ culture in the presence of a serum-free medium containing a mixture of growth factors (epidermal growth factor, insulin, pituitary extract, and dihydrotestosterone). In many of the tissues examined at 4 days, individual glands resembled those seen immediately after surgery, with a single layer of basal epithelial cells and a layer of secretory cells above. By Day 8, the secretory epithelium was lost in many places and basal cells proliferated to fill in the lumens of the glands. All of the nonmalignant glands were reactive with the anti-cytokeratin antibody (K903), and there was a large increase in the number of cells staining for Ki-67 as compared with zero-time tissue. Staining with the Periodic Acid Schiff (PAS) and PAS-methenamine silver (PASME) reagents revealed an intact basement membrane around virtually all of the epithelial structures. The basement membrane appeared to be thickened in some areas. In places where a gland was cut during the processing of the tissue, epithelial cells migrated out of the gland and covered the cut surface of the tissue piece. There was no detectable basement membrane separating the epithelium from the stroma at these sites. Whereas nonmalignant epithelial cells were preserved in the growth factor- and dihydrotestosterone-supplemented culture medium, most of the malignant cells rapidly lysed under the same conditions. However, when phorbol myristate acetate was included in the culture medium, many of the tumor cells remained viable. This was seen with the more well differentiated tumors as well as with tumors that were highly anaplastic. All of the tumor cells were nonreactive with anti-cytokeratin antibody, and only a few cells stained for Ki-67. The basement membrane surrounding malignant cells was thin and, in places, appeared to be discontinuous. Where malignant glands were cut in the processing of the tissue, cells did not migrate out over the cut surface. In summary, this study identifies culture conditions for the successful maintenance of human prostate tissue for several days in organ culture. Histological/histochemical features that distinguish nonmalignant and malignant tissue are present in this model. PMID- 10378516 TI - Fluorescence spectroscopic and histochemical analysis using hematoporphyrin as a microenvironmental probe for atherosclerotic change in the human aorta. AB - Hematoporphyrin (HP) was used as a microenvironmental fluorescent probe to investigate the development of human atherosclerotic plaques. We compared the site of HP accumulation with changes in the HP fluorescence spectrum in atheromatous plaques and in a liposome control model, using a confocal laser scanning microscope equipped with a photonic multi-channel analyzer linked to a fluorescence spectrometer. Wavelength shifts of the two peaks of HP fluorescence (F1, 620 nm; and F2, 640-690 nm) were monitored, and the integrated F2/F1 ratio was calculated as a measure of HP fluorescence. The F1 peak is characteristic of a predominantly aqueous site. The ratio reflects selective changes in the distribution and overcrowding of HP molecules in the limited space of the artificial membrane model. Compared with a normal artery, the atherosclerotic lesions showed an increase in the area of HP fluorescence, increased fluorescence intensity, a red shift of the HP fluorescence spectrum, and an increased F2/F1 ratio. The F2/F1 ratio of the membranous structures was markedly greater in the cores of the fibrous plaques than in the other plaques. The F1 peak showed increased intensity in the atheromatous plaque core, whereas in hydrophilic fibrous regions, such as the cap of the plaque, the intensity of the F1 peak was lower than in the core region. HP aggregation was observed in damaged cells and in water surrounded by lipid in the atheromatous core. Using HP as a probe allowed us to determine not only the ionization or polarity of each region in the atherosclerotic plaques but also to detect the separation and fusion of the lipid bilayer or micelle lipids, as well as damage to the cellular membranes and cholesterol enrichment. These findings suggest that HP is useful for detecting clinically important changes in atherosclerotic lesions, to lipid-rich, unstable, and vulnerable plaques, which are closely associated with cardiovascular events. PMID- 10378517 TI - CD44 variant isoforms on blood leukocytes in chronic inflammatory bowel disease and other systemic autoimmune diseases. AB - We have described recently that TNBS-induced colitis, an animal model of chronic inflammatory bowel disease (IBD), can be cured by treatment with anti-CD44v7. This finding led us to evaluate whether CD44v7 may be of functional importance in patients with IBD. Expression of CD44 variant isoforms (CD44v) has been evaluated on PBMC of 46 patients with IBD, 43 patients with autoimmune diseases not affecting the gastrointestinal tract, 26 patients with nonautoimmune disease, and 24 healthy donors. In all groups, expression of CD44v on freshly harvested PBMC was not above or was borderline above background levels. After in vitro stimulation, expression of CD44 standard (CD44s) and CD44v6 was strongly up regulated. Exclusively on PBMC of patients with autoimmune disease, high expression of CD44v3 and CD44v7 was observed. CD44v3 and CD44v7 were mainly expressed on subsets of CD4+ lymphocytes, B cells, and monocytes; CD44v6 was predominantly detected on CD4+ and CD8+ cells. Considering functional activity, CD44v7 apparently exerted a dual effect. After culturing PBMC in the presence of anti-CD44v7, a higher percentage of cells produced IL-10. This was irrespective of whether the PBMC were derived from healthy donors or from patients with autoimmune disease or IBD. On the other hand, PBMC of all donors proliferated upon cross-linking of CD3 and CD44s or CD3 and CD44v6. Instead, costimulatory activity of CD44v7 was seen only in PBMC of patients with autoimmune disease and IBD. Because expression and function of CD44v7 in patients with systemic autoimmune disease and IBD have been much like the ones in mice suffering of TNBS induced colitis, it is tempting to speculate that blockade of CD44v7 could also be of therapeutic relevance in the human diseases. PMID- 10378518 TI - Auditory screening in neonates by means of transient evoked otoacoustic emissions: a report of 2,842 recordings. AB - The principal goal of an early identification program is to identify hearing impairment present at birth, in order to effect appropriate intervention as early as possible. Although recent research provides some evidence for the value of transient evoked otoacoustic emissions (TEOAEs) in neonate hearing screening, data are needed from large-scale clinical evaluations about the value of using TEOAEs for screening not only high-risk but also healthy neonates. A cohort of 1,421 neonates (2,842 ears) from the well-baby nursery was screened with TEOAEs in a 2-stage process. Neonates were referred from the first test prior to being discharged from the hospital. Those who failed were rescreened before the end of the first month. Those who did not pass the second-stage TEOAE screening were referred for diagnostic audiological evaluation for confirmation of hearing loss. Neonates transferred to a neonatal intensive care unit were not included in this study. Two neonates with bilateral sensorineural hearing loss of >40 dB hearing level were identified from this cohort. This study demonstrates the feasibility and the limitations of using TEOAEs as a universal hearing screening tool for all neonates. It confirms that the prevalence of hearing impairment in neonates has to be taken into account, even in a group of children without high-risk criteria. In France, a prevalence of 1.4 per 1,000 would represent 1,000 deaf children born every year, with reference to about 700,000 births per year. This study suggests that such universal screening programs would substantially increase the rate of early-identified infants with significant hearing impairment. PMID- 10378519 TI - Percutaneous implants in the temporal bone for securing a bone conductor: surgical methods and results. AB - Clinical results of an extended follow-up of percutaneous titanium implants for application of bone conductors are presented. A simplified 1-stage surgical procedure is introduced. This study entails a consecutive series of 163 implants in 155 patients 9 to 80 years old who received a bone conductor coupled to a percutaneous titanium implant since June 1988 at the University Hospital Nijmegen. The maximum follow-up is 7 years. The bone conductor can be connected to a percutaneous abutment fixed to a titanium fixture anchored in the temporal bone. Several clinical trials from different clinics have shown its efficacy in patients with a conductive or mixed hearing loss. In this study attention is paid to the following issues: the occurrence of skin reactions, the condition of the skin around the abutment, the stability of the fixture, and a simplified surgical technique. PMID- 10378520 TI - Electron microscopic temporal bone histopathology in experimental pneumococcal meningitis. AB - Bacterial meningitis is one of the most common causes of acquired profound sensorineural deafness in children. Measurement of hearing and examination of the cochlea is limited in patients suffering from acute meningitis. A rabbit model of pneumococcal meningitis was developed to identify the temporal bone histopathologic changes that occur in meningogenic labyrinthitis caused by Streptococcus pneumoniae. Light microscopy was previously performed on temporal bones from acutely meningitic rabbits with profound hearing loss as determined electrophysiologically. Extensive inflammation of the cochlea with endolymphatic hydrops was observed. The organ of Corti, however, showed preserved architecture in the majority of these animals. In order to further investigate these findings, a protocol was used to create meningitic rabbits with hearing loss ranging from early high-frequency loss to profound deafness. The temporal bones from 7 rabbits were examined by transmission electron microscopy. In cases of mild hearing loss, partial degeneration of the inner row of outer hair cells, as well as edema of efferent cochlear nerve endings and marginal cells of the stria vascularis, was seen. With increasing degrees of hearing loss, the remainder of the organ of Corti and intermediate cells of the stria showed ultrastructural abnormalities. Spiral ganglion cells and basal cells of the stria vascularis remained intact in all subjects. This study provides unique information regarding the histology and pathophysiology of meningogenic deafness. The clinical implications of these findings are discussed, with an emphasis on potentially reversible changes and therapeutic intervention. PMID- 10378521 TI - Relationship of the utriculus and sacculus to the stapes footplate: anatomic implications for sound-and/or pressure-induced otolith activation. AB - One hundred thirty human temporal bones that were sectioned in the vertical plane were examined to evaluate the relationship between the stapes footplate and the otolith organs. The shortest distance between the footplate and the utriculus was 0.58+/-0.10 mm in the posterior third of the oval window, 1.04+/-0.20 mm in the middle third, and 1.51+/-0.20 mm in the anterior third. The distance from the sacculus to the footplate was 1.33+/-0.20 mm in the middle third of the oval window and 1.31+/-0.18 mm in the anterior third. Membranous connections extending between the utriculus and the footplate were found in 26% of temporal bones. These membranous connections in coexistence with additional anatomic factors such as stapes hypermobility and/or dehiscence of bone within labyrinthine structures may predispose patients to sound- and/or pressure-induced otolith activation. The findings may have implications for different causes of the Tullio phenomenon. PMID- 10378522 TI - Efficacy of an antiallergic drug on otitis media with effusion in association with allergic rhinitis. An experimental study. AB - The effect of an antiallergic drug on the evacuation of middle ear effusion (MEE) from the tubotympanum was investigated by means of an animal model with both otitis media with effusion (OME) and allergic rhinitis. Azelastine hydrochloride (AZ), an oral antiallergic drug, was administered and the presence of MEE was investigated. Serous MEE was seen in 12 of the 13 untreated control animals on the 11th day after the experimental OME was induced, whereas MEE was detected in 9 of the 13 animals administered 1 mg/kg of AZ, but only in 4 of the 13 animals administered 2 mg/kg of AZ. In addition, the effect of AZ on MEE production was also examined in an experimental OME animal model without allergic rhinitis. Middle ear effusion was observed in all OME animals that received 2 mg/kg AZ for 5 consecutive days, before and 3 days after the experimental OME was induced. Results of the present study indicate that AZ promotes the evacuation of MEE from the tubotympanum in the OME animal model associated with nasal allergy. These data suggest that an antiallergic drug may contribute to the therapy of OME patients in association with nasal allergy indirectly, by promoting evacuation of MEE due to inhibition of type I allergic reactions in the nasopharynx. PMID- 10378523 TI - Course of IL-1beta, IL-6, IL-8, and TNF-alpha in the middle ear fluid of the guinea pig otitis media model induced by nonviable Haemophilus influenzae. AB - To characterize the local response in acute otitis media, courses of interleukin (IL)-1beta, IL-6, IL-8, and tumor necrosis factor (TNF)alpha in middle ear fluid (MEF) of the guinea pig otitis media model induced by nonviable Haemophilus influenzae were investigated with enzyme-linked immunosorbent assay (ELISA) kits. The IL-1beta concentration in H. influenzae-inoculated ears peaked 24 hours after inoculation. The IL-8 concentration was significantly higher in H. influenzae inoculated ears than in controls 48 and 96 hours after inoculation. The TNF-alpha concentration in H. influenzae-inoculated ears had an initial peak 6 hours after inoculation and had significant late increases 48 and 96 hours after inoculation. The results suggest that IL-1beta and TNF-alpha were produced by middle ear mucosa in the early stage of the experiment by stimulation of bacterial inoculation, which caused subsequent inflammatory cell accumulation, and that IL 8 and TNF-alpha were produced in the late stage by accumulating inflammatory cells. PMID- 10378524 TI - Lipid peroxides in middle ear fluid after acute otitis media in guinea pigs. AB - Oxygen free radical damage has been demonstrated in the middle ear mucosa of a guinea pig model of acute otitis media (AOM). Potential sources of free radicals include both neutrophils responding to infection and Streptococcus pneumoniae, a common AOM pathogen. This project was conducted to examine the middle ear fluid in a guinea pig model of AOM for evidence of elevated lipid peroxide (LPO) as a marker of free radical damage. Twenty-one guinea pigs were injected transtympanically with bacteria into the left (infected) middle ear cavity and sterile saline into the right (control) middle ear. Middle ear fluid was recovered on postoperative day 5. The fluid was weighed and analyzed for quantity of LPO. Results indicated an increased absolute level of LPO, as well as an increased level of LPO divided by the weight of the fluid recovered. Histologic examination confirmed leukocyte infiltration and mucosal edema that were consistent with mucosal damage. While free radical damage to the middle ear mucosa in a guinea pig model of AOM is well documented, this is the first study to demonstrate evidence of free radical damage in middle ear fluid. These results are relevant because they correlate mucosal damage with lipoperoxidation in fluid. Additionally, this serves as an important precursor to human studies, since middle ear fluid is readily available in patients with otitis media. PMID- 10378525 TI - Treatment of hypertrophy of the inferior turbinate: long-term results in 382 patients randomly assigned to therapy. AB - A number of surgical techniques are commonly performed to control the symptoms of inferior turbinate hypertrophy unresponsive to medical treatment. We report long term results in 382 patients randomly assigned to receive electrocautery (62), cryotherapy (58), laser cautery (54), submucosal resection without lateral displacement (69), submucosal resection with lateral displacement (94), and turbinectomy (45). Outcomes of objective test results from rhinomanometry, acoustic rhinometry, mucociliary transport time, and secretory immunoglobulin A levels were compared to the symptom scores before and yearly after surgical treatment. These data indicate that submucosal resection with lateral displacement of the inferior turbinate results in the greatest increases in airflow and nasal respiratory function with the lowest risk of long-term complications. PMID- 10378526 TI - Abnormal expression of the cystic fibrosis transmembrane regulator in chronic sinusitis in cystic fibrosis and non-cystic fibrosis patients. AB - Cystic fibrosis (CF) patients commonly suffer from chronic sinusitis. Mutations of a single gene, the cystic fibrosis transmembrane conductance regulator (CFTR) gene, have been associated with CF. Functional CFTR protein is localized to the apical cell membrane, while dysfunctional CFTR is commonly found in the cytoplasm. We undertook a preliminary immunocytochemical study of CFTR subcellular localization in CF and non-CF pediatric and adult patients using a newly developed murine monoclonal antibody, TAM. Immunostaining was evaluated for subcellular localization (cytoplasmic versus membranous) and for epithelial layer (basal versus luminal). Analysis of the predominant CFTR distribution patterns demonstrated significant differences in adult versus pediatric groups independent of whether the latter were CF or non-CF (p<.0001 and p<.008, respectively), and no significant difference between the 2 pediatric groups (p = .70). This suggests that the pathophysiology of pediatric sinusitis differs from that of adult sinusitis at the level of secretion production. PMID- 10378527 TI - Effect of diesel exhaust on guinea pig nasal mucosa. AB - In this study using guinea pigs, we investigated the effects of diesel exhaust (DE) containing diesel exhaust particulate (DEP) on 1) vascular permeability induced by histamine, 2) nasal mucosal permeability to horseradish peroxidase (HRP), and 3) eosinophilic epithelial infiltration. The vascular permeability induced by histamine was enhanced significantly and dose-dependently in DE exposed guinea pigs. The HRP reaction products in epithelial cells and intercellular spaces were significantly and dose-dependently increased in those guinea pigs. Eosinophil infiltration into the epithelial layer was significantly increased in guinea pigs exposed to DE containing 3.2 mg/m3 DEP, and the reactivity of the nasal mucosa to histamine solution applied on the nasal mucosa was significantly enhanced in those guinea pigs. These findings suggest that DE may play an important role not only in promoting nasal hyperreactivity induced by the enhancement of absorption of antigen through the nasal epithelium, but also in inducing eosinophil infiltration in nasal mucosa and enhancing nasal mucosal reactivity. PMID- 10378528 TI - Congenital nasal hemangiopericytoma: intrauterine, intraoperative, and histologic findings. AB - Hemangiopericytoma is a rare tumor of mesenchymal origin. To date, 91 cases of nasal or paranasal hemangiopericytoma and 59 congenital hemangiopericytomas have been reported in the literature. A congenital hemangiopericytoma arising from the nasal cavity and skull base has not yet been described. We report a case of a male newborn with a highly vascular nasal tumor diagnosed by in utero sonography with three-dimensional surface reconstruction. The tumor extended to the right anterior skull base, the right nasal cavity, and the right side of the nasal pyramid. A complete resection by neodymium:yttrium-aluminum-garnet-potassium titanyl phosphate ("Nd:YAG-KTP") laser was performed on the day of cesarean section at 33 weeks' gestation. The tumor was diagnosed as hemangiopericytoma by histologic and immunohistochemical findings. Postoperative nasal flow, feeding, and sight were unimpaired. At the 9-month follow-up, the infant remained free of disease. PMID- 10378529 TI - Distribution of rantes and interleukin-5 in allergic nasal mucosa and nasal polyps. AB - Eosinophil-chemoattracting cytokines are thought to be important in the pathogenesis of allergic inflammation. However, little is known about the presence and significance of RANTES in nasal allergy and nasal polyps, two well known rhinologic disorders characterized by eosinophil infiltration in the tissue. In order to evaluate the role of RANTES in eosinophil infiltration in vivo, the tissue distributions of RANTES and interleukin-5 (IL-5) and their correlation with eosinophil infiltration were investigated. Nasal mucosa specimens were obtained from 9 allergic and 12 control subjects, and nasal polyps from 6 allergic and 9 nonallergic subjects. All the subjects were divided into 4 groups: normal mucosa, allergic mucosa, nonallergic polyps, and allergic polyps. To identify the cellular localizations of RANTES and IL-5, we used specific immunohistochemical staining. We also investigated the differences in cytokine expression among the 4 groups, and the correlation between cytokine expression and eosinophil infiltration in the tissue. RANTES was expressed in the epithelium, endothelium, and some submucosal cells, while IL-5 was confined to the cells in the submucosa. Expression of both RANTES and IL-5 significantly increased in allergic mucosa and nasal polyps compared to normal mucosa; however, there was no significant difference in their expression between allergic and nonallergic polyps. Both cytokines had a significant correlation between their expression and either total or activated eosinophil numbers. The results of this study suggest that RANTES, as well as IL-5, plays a role in eosinophil recruitment in allergic nasal mucosa and nasal polyps in vivo. PMID- 10378530 TI - Graft healing in laryngotracheal reconstruction: an experimental rabbit model. AB - The actual sequence of events in graft healing in laryngotracheal reconstruction (LTR) is not well understood. To investigate sequential changes in graft healing, we submitted 21 rabbits to LTR with an anterior auricular cartilage graft. They were immediately extubated after surgery, and no stent was used. Rabbits were painlessly sacrificed at 7 different time periods (0, 1, 2, 3, 4, 6, and 10 weeks after surgery). Cross sections of the larynx and graft were cut and stained with hematoxylin-eosin and trichrome. Epithelialization progressed very rapidly and was complete by 21 days. During the 10 weeks, there was a progressive necrosis and resorption of the original graft cartilage. However, neochondrification progressed very rapidly and resulted in excellent structural support of the graft in the airway. Problems, such as infection and granulation tissue formation, were relatively minor and infrequent. PMID- 10378531 TI - Swallowing disorders in paralysis of the lower cranial nerves: a functional analysis. AB - Deficits of the lower cranial nerves (nerves IX, X, XI, and XII) occurring after treatment of skull base tumors may cause disabling swallowing disorders. To assess the mechanisms of swallowing disorders involved in such cases, we performed functional examinations: a videoendoscopic swallowing study and simultaneous manometry and videofluoroscopy in 7 patients. This study shows that the main mechanism of the swallowing disorders was a disturbance of the pharyngeal stage, including a decrease of pharyngeal propulsion, reduced laryngeal closure, and cricopharyngeal dysfunction, which led to aspiration. Decreased pharyngeal propulsion was found in 6 patients, with a very high correlation between fiberoscopy and simultaneous manometry-fluoroscopy. The responsibility of the upper esophageal sphincter in swallowing disorders was more difficult to assess. The role of the upper esophageal sphincter and pharyngeal propulsion in the onset of the problem is discussed in regard to the cricopharyngeal myotomy. PMID- 10378533 TI - High jugular bulb adhering to the eardrum. PMID- 10378532 TI - Laryngeal long latency response conditioning in abductor spasmodic dysphonia. AB - Previously, we demonstrated that patients with adductor spasmodic dysphonia (ADSD) have a disinhibition of laryngeal responses to sensory input. In this study, sensorimotor responses to stimulation of the superior laryngeal nerve were compared between 10 subjects with abductor spasmodic dysphonia (ABSD) and 15 normal volunteers. The groups had similar latency and frequency characteristics of their unconditioned adductor responses (p>.05). The conditioned R1 (early) responses of the subjects with ABSD were greater and more variable in amplitude than those of the normal volunteers (p< or =.008). Similar R2 (late) conditioning effects were found in both groups, with a nonsignificant trend toward reduced inhibition of contralateral R2 responses at lower interstimulus intervals (p = .01) in the patient group. Thus, inhibitory mechanisms that modulate the R1 laryngeal sensorimotor pathway in the brain stem may be abnormal in subjects with ABSD. Abnormal modulation of laryngeal sensorimotor responses seems present in both types of spasmodic dysphonia. PMID- 10378534 TI - Detection of unknown primary cancer with fluor-deoxy-glucose positron emission tomography. PMID- 10378535 TI - Follow-up of patients with completely resected lung cancer. PMID- 10378536 TI - Cytokines and pleural drainage fluid: do local levels make a difference? PMID- 10378537 TI - Improving outcomes in the ICU setting: are we effectively using all of the information that is potentially available to us? PMID- 10378538 TI - A stuck pig--even on warfarin--doesn't always bleed. PMID- 10378539 TI - Follow-up in lung cancer: how often and for what purpose? AB - OBJECTIVES: The present study evaluates the cost-effectiveness of two follow-up routines: a strict follow-up with frequent visits, imaging, and laboratory examinations was compared to a follow-up with infrequent visits that were scheduled mainly on the basis of the patient's symptoms. METHODS: A retrospective evaluation was undertaken of 130 patients who underwent a complete resection of non-small cell lung cancer (NSCLC). All patients had complete follow-up for at least 2 years after their operation. The patients were separated into two groups: strict (n = 67), with a routine follow-up policy; and symptom (n = 63), seen on a symptom-oriented basis. The costs of the follow-up routines and the yield of each schedule were compared between the two groups. RESULTS: There were no significant differences in the disease-free interval until the first detection of recurrence. In most patients, metastatic diseases were diagnosed on the basis of symptoms, rather than by routine tests. The patients who had recurrent cancer diagnosed after surgery had a dismal survival rate irrespective of the follow-up schedule. The majority of patients with recurrence died of malignancy within a 2-year period. The costs of strict vs symptom follow-up were significantly different, because of the greater number of routine imaging procedures performed in patients having strict follow-up. On the other hand, when we analyzed only the frequency of hospitalization and the cost per day of hospital treatment for medical problems other than cancer recurrence, the patients in the strict group had a less expensive follow-up than the patients in the symptom group. CONCLUSIONS: The present study showed that a more cost-effective routine follow-up scheme should be advised for patients with completely resected NSCLC, without affecting overall outcome. Routine imaging follow-up is of questionable value, and it may be indicated only in academic settings. PMID- 10378540 TI - Assessment of usefulness of endobronchial ultrasonography in determination of depth of tracheobronchial tumor invasion. AB - STUDY OBJECTIVE: We assessed the usefulness of endobronchial ultrasonography in the determination of the depth of tumor invasion of the tracheobronchial wall. METHODS: We performed a needle-puncture experiment on normal tissue of 45 specimens to determine the laminar structure of the tracheobronchial wall. In addition, we compared the ultrasonographic determinations of tumor invasion from 24 lung cancer cases with the histopathologic findings. RESULTS: The cartilaginous portions of the extrapulmonary bronchi and the intrapulmonary bronchi exhibited a five-layer structure. Starting on the luminal side, the first layer (hyperechoic) was a marginal echo, the second layer (hypoechoic) was the submucosal tissue, the third layer (hyperechoic) was the marginal echo on the inner side of the bronchial cartilage, the fourth layer (hypoechoic) was bronchial cartilage, and the fifth layer (hyperechoic) was the marginal echo on the outer side of the cartilage. In the membranous portions, the first layer (hyperechoic) was a marginal echo, the second layer (hypoechoic) was smooth muscle, and the third layer (hyperechoic) corresponded to the adventitia. Comparisons between the ultrasonograms and the histopathologic findings in 24 lung cancer cases revealed that depth diagnosis was the same in 23 lesions (95.8%) and was different in 1 lesion (4.2%). In the single case in which the findings were different, lymphocytic infiltration that protruded between the cartilage rings was mistakenly interpreted as tumor infiltration. CONCLUSIONS: This method allows visualization of the laminar structure of the tracheobronchial wall, which is impossible with other diagnostic imaging methods. PMID- 10378542 TI - Combined effects of a mechanical nasal dilator and a topical decongestant on nasal airflow resistance. AB - The goal of this study was to compare the isolated and combined effects of two treatments being used to reduce nasal airflow resistance (NR): an internal nasal mechanical dilator (Nozovent; Prevancure; Ste Pouret, Paris, France) and a topical decongestant, fenoxazoline hydrochloride (Aturgyl; Synthelabo; Le Plessis Robinson, France). The study was performed in 17 healthy subjects. NR was estimated by active posterior rhinometry at a 0.5 L/s flow under four conditions: in the basal state, with the internal nasal mechanical dilator, after treatment with fenoxazoline hydrochloride, and with both fenoxazoline hydrochloride and the mechanical dilator. The mean NR (+/- SD) decreased from 1.65+/-0.54 cm H2O/L/s in the basal state to 1.02+/-0.27 cm H2O/L/s with the mechanical dilator (p < 0.001), 1.03+/-0.47 cm H2O/L/s with fenoxazoline hydrochloride (p < 0.001), and 0.48+/-0.15 cm H2O/L/s with both the mechanical dilator and fenoxazoline hydrochloride (p < 0.001). The decreases in NR observed after using either the mechanical dilator (deltaNR(N)) or fenoxazoline hydrochloride (deltaNR(A)) were not significantly different. The decrease in NR observed with both (deltaNR(N + A)) was not significantly different from the sum deltaNR(N) + deltaNR(A): 1.16+/ 0.53 cm H2O/L/s vs 1.25+/-0.63 cm H2O/L/s, respectively (p > 0.05). deltaNR(N + A) strongly correlated with deltaNR(N) + deltaNR(A): deltaNR(N + A) = 0.80 (deltaNR(N) + deltaNR(A)) + 0.15 (r = 0.96; p < 0.0001). However, the slope of the regression line of deltaNR(N + A) vs deltaNR(N) + deltaNR(A) was significantly lower than unity (p < 0.003). These results demonstrate that, although not totally additive, the effects of using the mechanical dilator and fenoxazoline hydrochloride are cumulative. Further studies that include patients with nasal obstruction would allow us to better evaluate the benefit of a therapy combining both treatments. PMID- 10378541 TI - Impact of revised stage classification of lung cancer on survival: a military experience. AB - STUDY OBJECTIVES: This report reviews results of surgical management of lung cancer at a military medical center using the revised 1997 stage classification and determines the impact of the revised system on survival rates. It also compares our results with the recent reports from Japan and from a large, multinational study involving several institutions. DESIGN: Retrospective review. SETTING: Department of Cardiothoracic Surgery, Walter Reed Army Medical Center (WRAMC), Washington, DC. PATIENTS OR PARTICIPANTS: Active military members, their dependents, and eligible retired military members who were admitted to WRAMC for surgical treatment of lung cancer between January 1984 and December 1996. METHODS: Records of all patients who had surgical resection with intent to cure were reviewed. Data extracted included clinical and pathologic stages according to the 1997 revised stage classification. Survival probabilities for the stages were calculated by the Kaplan-Meier actuarial method. The log rank test was used to compare survival rates between stages and stage subsets. A p value < 0.05 was considered statistically significant. MEASUREMENTS AND RESULTS: Five hundred fifty-two of the 1,398 patients with primary lung cancers underwent curative surgical resection (39.5%). The operative mortality was 2%. Using the revised 1997 stage classification, the survival rate for stage IA was 77%; IB, 62%; IIA, 57%; IIB, 47%; IIIA, 28%; IIIB, 20%; and IV, 0%. The overall actuarial 5-year and 10-year survival rates were 58% and 45%, respectively (median survival, 3.3 years; mean survival 3.9+/-0.1 years). CONCLUSIONS: Our results confirm the justification for the recent revisions in the staging system of lung cancer; however, there are still discrepancies that cannot be explained. PMID- 10378543 TI - Comparison of respiratory polysomnographic parameters in matched cohorts of upper airway resistance and obstructive sleep apnea syndrome patients. AB - OBJECTIVE: To compare respiratory nocturnal polysomnography (NPSG) characteristics between matched cohorts of upper airway resistance syndrome (UARS) and obstructive sleep apnea syndrome (OSAS) patients. METHODS: All patients received 13-channel NPSG, including esophageal pressure (Pes) manometry. By definition, OSAS patients had an apnea-hypopnea index (AHI, number of apneas/hypopneas per hour total sleep time) > or = 15, and UARS patients had an AHI < 5. Respiratory effort-related arousal (RERA) was defined as the absence of apnea/hypopnea with > or = 10 s duration of progressive negative Pes, culminating in an arousal or microarousal. UARS patients, by definition, had > or = 15 RERAs per hour. Fifteen consecutively diagnosed UARS patients were matched with OSAS patients on the basis of body mass index (BMI) and gender. RESULTS: Respiratory disturbance index (sum of the AHI and RERA per hour) was the same for both cohorts: UARS, 36+/-4; OSAS, 42+/-6 (p = 0.34). There were no differences between cohorts for mean inspiratory Pes nadirs for each 30-s epoch of sleep compared for each sleep stage over an entire night. For randomly selected breaths from supine stage 2 sleep, the mean inspiratory Pes nadir was the same for the cohorts: UARS, -16.6+/-2 cm H2O; OSAS, -16.1+/-3 cm H2O (p = 0.30). Differences between cohorts for each parameter fell within respective 95% confidence intervals. CONCLUSION: With the exception of AHI, respiratory NPSG parameters were the same for UARS and OSAS patients when BMI and gender were controlled for. PMID- 10378544 TI - Salmeterol vs theophylline: sleep and efficacy outcomes in patients with nocturnal asthma. AB - STUDY OBJECTIVES: To compare the efficacy, safety, and effects on sleep quality of salmeterol and extended-release theophylline in patients with nocturnal asthma. DESIGN: Randomized, double-blind, double-dummy, three-period crossover. SETTING: Outpatients at a single center. Patients spent 1 night during screening and 2 nights during each study period in a sleep laboratory for completion of sleep studies. PATIENTS: Male and female patients who were at least 18 years old with nocturnal asthma (baseline FEV1, 50 to 90% of predicted) and who required regular bronchodilator therapy. Patients on inhaled corticosteroids, cromolyn, and nedocromil were allowed into the study if their dosing remained constant throughout the study. INTERVENTIONS: Inhaled salmeterol (42 microg per actuation), extended-release oral theophylline (titrated to serum levels of 10 to 20 microg/mL), and placebo taken twice daily. MEASUREMENTS AND RESULTS: Efficacy measurements included nocturnal spirometry, nocturnal polysomnography, sleep questionnaires, and daily measurements of lung function and symptoms. Salmeterol was superior to theophylline (p < or = 0.05) in maintaining nocturnal FEV1 levels and was superior to placebo (p < or = 0.05) in improving morning and evening peak expiratory flow (PEF) and in decreasing nighttime albuterol use. The use of salmeterol significantly increased the percentage of days and nights with no albuterol use and decreased daytime albuterol use compared with theophylline and placebo (p < or = 0.05). Sleep quality global scores significantly improved with salmeterol and placebo (p < 0.001) but not with theophylline. The effects on sleep architecture were similar across treatment groups. CONCLUSIONS: Salmeterol (but not theophylline) was associated with sustained improvements in morning PEF, protection from nighttime lung function deterioration, reductions in albuterol use, and improvements in patient perceptions of sleep. No differences were seen in polysomnographic measures of sleep quality. PMID- 10378545 TI - Effects of a beta2-agonist on airway hyperreactivity in subjects with cervical spinal cord injury. AB - STUDY OBJECTIVE: Aerosolized ipratropium bromide or orally administered baclofen or oxybutynin chloride (Ditropan) block methacholine-associated airway hyperreactivity in subjects with chronic cervical spinal cord injury (SCI), whereas these agents do not inhibit airway hyperreactivity associated with the inhalation of histamine. The present study was performed to determine whether pretreatment with a beta2-agonist attenuates airway hyperresponsiveness in these subjects. PARTICIPANTS: Subjects with chronic cervical SCI previously demonstrating airway hyperreactivity were challenged with methacholine (n = 9) or histamine (n = 16) alone and, on a separate day, 25 min following inhalation of nebulized metaproterenol sulfate. RESULTS: Inhalation of the beta2-agonist was associated with an increase in provocative concentration causing a 20% decrease in FEV1 (PC20) values (geometric mean) from 1.01+/-2.76 to 20.54+/-6.24 mg/mL for methacholine and from 2.29+/-2.26 to 19.82+/-5.93 mg/mL for histamine. No correlation was found between specific PC20 values for individual subjects and percentage improvement in FEV1 (liter) following inhalation of metaproterenol sulfate and between PC20 values and baseline FEV1 percent. CONCLUSION: These data, combined with findings that patients with chronic high cervical SCI experience increased breathlessness following exposure to exogenous agents, suggest that long-term prophylactic beta2-agonist therapy may reduce respiratory symptoms associated with airway hyperreactivity in these patients. PMID- 10378546 TI - Micturitional disturbances are associated with impaired breathing control in multiple sclerosis. AB - STUDY OBJECTIVES: To investigate whether the localization of multiple sclerosis (MS), the duration of the disease, and the level of neurologic functioning in patients with MS predispose them to disturbed breathing control. DESIGN: Case control study. SETTING: Outpatient pneumology department of a university hospital. PATIENTS: Twenty-three MS patients and 51 healthy control subjects. MEASUREMENTS AND RESULTS: Resting mouth occlusion pressure at 0.1 s after onset of inspiratory effort (P0.1) was measured during the hypercapnic response (HCR) and the hypoxic response (HR) in all subjects. The Kurtzke expanded disability status scale and the functional system score were used to describe the level of neurologic functioning of the MS patients. Predictors of HCR and HR were assessed by multiple regression analysis. Low maximal inspiratory pressure (MIP) values correlated with low resting P0.1 values (r = 0.44; p = 0.05), although in neuromuscular diseases, high resting P0.1 values are usually found to compensate for low MIPs. Detrusor-sphincter dyssynergia (DSD) was the only predictor for lower ventilatory HCR (p = 0.006; r2 = 0.52), lower P0.1 HCR (p = 0.004; r2 = 0.47), lower ventilatory HR (p = 0.04; r2 = 0.28), and lower P0.1 HR (p = 0.04; r2 = 0.10); low MIPs and pyramidal tract involvement had no role. CONCLUSIONS: (1) Impaired control of breathing in some MS patients is related mainly to central defects. (2) DSD is the most important predictor of disturbed ventilatory control, presumably because the micturition and pneumotaxic center are closely related and located in the rostral pons. (3) No relationship with the duration of the MS disease could be demonstrated, which can be explained by the variable course of MS itself. PMID- 10378547 TI - Reliability of maximal respiratory pressures in multiple sclerosis. AB - OBJECTIVE: To assess the reliability of maximal inspiratory pressure (P(I)max) and maximal expiratory pressure (P(E)max) in subjects with multiple sclerosis (MS) and healthy control subjects by identifying the number of testing sessions and the number of measurements needed in a single testing session to obtain consistent, reproducible results. DESIGN: A descriptive, comparative design with repeated measures was used. SETTING: Four sets of 10 P(I)max and 10 P(E)max measurements were obtained over a 4-week period from MS subjects in their homes. The same measurements were obtained from healthy control subjects in a private setting. SUBJECTS: Seventy-two MS patients and 61 healthy control subjects participated in the study. MEASUREMENT: P(I)max and P(E)max values were obtained by using previously published methods. RESULTS: Mean P(E)max and P(I)max values for MS patients differed over the first three of the four testing sessions. By contrast, mean P(E)max and P(I)max values for healthy control subjects differed only when the first session values were compared with values from the last three sessions. For MS patients, P(E)max and P(I)max increased between the first and 10th trial during the first testing session, but not during the subsequent three sessions. CONCLUSIONS: The results of this study suggest that several practice sessions should be provided in order to obtain reliable P(E)max and P(I)max values in persons with MS. At least one practice session should be provided for healthy control subjects before identifying a baseline. PMID- 10378548 TI - Respiratory muscle function and hypoxic ventilatory control in patients with type I diabetes. AB - STUDY OBJECTIVES: The interaction among pulmonary mechanics, respiratory muscle performance, and ventilatory control in subjects with insulin-dependent diabetes mellitus has so far received little attention. We therefore decided to assess the role of central factors and peripheral factors on the ventilatory response to a hypoxic stimulus in type I diabetic patients. SUBJECTS: Eight patients in stable condition aged 19 to 48 years old, with insulin-dependent diabetes mellitus (duration of the disease, 36 to 240 months) and no history of smoking, cardiopulmonary involvement, or autonomic neuropathy; and an age- and gender matched control group. MEASUREMENTS: In each patient, we measured the following: pulmonary volumes; diffusing capacity of the lung for carbon monoxide (D(LCO)); time and volume components of ventilation (tidal volume [V(T)] and respiratory frequency); static compliance (Clstat) and dynamic compliance (Cldyn); swings in pleural pressure (Pes) and gastric pressure (Pg); and transdiaphragmatic pressure (Pdi), obtained by subtracting Pes from Pg. Maximal inspiratory Pes and Pdi during a maximal sniff maneuver were also measured. Swings in Pes and Pdi during V(T) as a percentage of Pes and Pdi during the maximal sniff maneuver [Pessw(%Pessn) and Pdisw(%Pdisn), respectively] were both considered as a measure of central respiratory output, and the Pessw(%Pessn)/V(T) ratio was considered as an index of neuroventilatory dissociation (NVD) of the inspiratory pump. Subjects were studied at baseline and during hypoxic rebreathing. RESULTS: Pulmonary volumes and D(LCO) were normal or slightly reduced. A lower Cldyn, higher central respiratory output, and NVD were found. During hypoxic rebreathing, patients had lower V(T), similar central respiratory output, and greater NVD per unit change in arterial oxygen saturation compared with values in control subjects. An increase in dynamic elastance, computed as 1/Cldyn, during hypoxia was found in patients, but not in normal subjects, and was directly related to concurrent changes in NVD. CONCLUSIONS: We have shown that the assessment of a normal Clstat and normal routine parameters of airway obstruction does not permit the definite exclusion of the role of peripheral airway involvement in insulin-dependent diabetes mellitus. Peripheral airway involvement is likely to influence indices of hypoxic ventilator) drive by modulating a normal central motor output into a rapid and shallow pattern of ventilatory response. PMID- 10378549 TI - The relation between physician experience and patterns of care for patients with AIDS-related Pneumocystis carinii pneumonia: results from a survey of 1,500 physicians in the United States. AB - STUDY OBJECTIVES: To determine whether physician experience and specialty influence the approach to care of AIDS patients with pneumonia, we surveyed physicians about their management of possible Pneumocystis carinii pneumonia (PCP) infection. DESIGN, SETTING, PARTICIPANTS: A postal survey was sent to a random sample of 1,500 internists and family physicians in the United States drawn from the American Medical Association master file who were identified by a pharmaceutical marketing company as having written prescriptions for AIDS-related agents in the previous year. MEASUREMENTS AND RESULTS: The survey had a 53% response rate. Physicians more experienced in AIDS care were more likely to advocate diagnostic bronchoscopy over initiation of empiric anti-PCP therapy for HIV-infected patients with undiagnosed pulmonary infiltrates (odds ratio [OR], 1.4 for a patient with mild severity of illness [p = 0.02]; OR, 1.7 for a severely ill patient [p < 0.001]). Physician specialty and fee-for-service reimbursement were independently associated with higher rates of bronchoscopy, with internists favoring bronchoscopy more frequently than family physicians. High-experience providers and internists also predicted better clinical outcomes for the hypothetical patients. CONCLUSIONS: Our findings extend the observations about HIV experience and PCP prophylaxis to the setting of diagnosis and treatment. Physicians with higher levels of experience with AIDS, internists, and physicians reimbursed as fee-for-service providers are more likely to support diagnostic confirmation of PCP than empiric treatment approaches. PMID- 10378550 TI - Oropharyngeal Gram-negative bacillary carriage: a survey of 120 healthy individuals. AB - BACKGROUND: The presence of aerobic Gram-negative bacilli (AGNB) in the oropharynx can be either temporary or persistent. Prolonged colonization (ie, carriage) is distinguished from transient presence (ie, acquisition), which often occurs in healthy individuals but less frequently in those with underlying disease. Prevalence rates of up to 61.1% quoted previously for healthy individuals were obtained by using single sample surveys, which fail to differentiate acquisition from carriage. STUDY OBJECTIVES: To illustrate the need to distinguish carriage from acquisition in a healthy population at risk of acquisition of AGNB, and to show that although differing groups of healthy individuals may acquire oropharyngeal AGNB at differing frequencies, carriage is rare in healthy individuals. PARTICIPANTS: Two oral rinses were obtained within a 2-day interval from 120 healthy individuals comprising 40 nurses, 40 students, and 40 laboratory-associated persons. DESIGN: Two hundred forty oral rinses were quantitatively (1:10 dilution series) cultured for AGNB by using broth enrichment. MEASUREMENTS AND RESULTS: The rate of AGNB carriage based on two consecutive samples positive for the same AGNB was 6.6%; the rate of AGNB acquisition based on one positive sample was 35.8%. The concentrations of all carried and acquired AGNB were < or = 103 cfu/mL. AGNB acquisition was significantly higher in students (52.5%) compared to nurses (32.5%) and laboratory-associated persons (22.5%; p < 0.05). CONCLUSION: Healthy individuals rarely carry oropharyngeal AGNB, suggesting effective oropharyngeal clearance in a healthy population predisposed to acquisition. Apparently, the oropharyngeal mucosa in healthy individuals is not receptive to adhesins of AGNB, resulting in rapid elimination of these bacteria. PMID- 10378551 TI - Infusion phlebitis in patients with acute pneumonia: a prospective study. AB - STUDY OBJECTIVES: To prospectively assess the relative risk for phlebitis in a series of consecutive patients with pneumonia and to identify risk factors that predict an increased risk for phlebitis. SETTING: Internal medicine department of a tertiary teaching hospital. PATIENTS: Seven hundred sixty-six consecutive patients with acute pneumonia receiving IV therapy. INTERVENTIONS: Only the first catheter was taken into account. There were 308 short lines (a 51-mm, 18-gauge Teflon catheter); 307 midsized lines (a 28-cm, 16-gauge polyvinyl chloride catheter); and 151 long lines (71-cm, 14-gauge plain polyurethane catheter). Eighteen variables were prospectively evaluated in an open, nonrandomized study for their contribution to the occurrence of phlebitis. RESULTS: The overall phlebitis rate was 39%. Phlebitis developed in 53% of patients with short lines, in 41% of patients with midsized lines, and in 10% of patients with long lines, and these catheters remained in place an average (+/- SD) of 3.0+/-2.4 days, 4.6+/-3.4 days, and 7.8+/-6.6 days, respectively. The variables that influenced the development of phlebitis, as determined by multivariate analysis, were the following: type of catheter; blood hemoglobin levels; and IV therapy with either corticosteroids or erythromycin. CONCLUSIONS: According to our data, when the use of a catheter is expected to be required for < or = 36 h, a short line can be used. If a longer duration is expected, a longer line is warranted. Ours is the first study in which the relationship between blood hemoglobin levels and phlebitis has been reported. Because the use of intravascular devices is increasingly common, a more complete knowledge of the factors that influence their acceptance has become essential. PMID- 10378552 TI - Respiratory insufficiency in postmenopausal women: sustained improvement of gas exchange with short-term medroxyprogesterone acetate. AB - STUDY OBJECTIVES: The degree and duration of respiratory stimulation of medroxyprogesterone acetate (MPA) in postmenopausal women. DESIGN: A placebo controlled single-blind trial. SETTING: University hospital in Turku, Finland. PATIENTS: Fourteen postmenopausal women with permanent or previous episodic hypercapnic or hypoxemic respiratory failure. INTERVENTIONS: A 12-week trial including 14-day treatment periods with placebo and MPA (60 mg daily) and a 6 week follow-up. RESULTS: Thirteen of 14 patients completed the trial. The mean (+/- SD) PaCO2 at baseline was 42.8+/-4.5 mm Hg and the mean PaO2 was 71.2+/-9.0 mm Hg. The average reduction of PaCO2 was 6.3 mm Hg (14.7%, p < 0.001) on MPA and 3.0 mm Hg (6.1%, p = 0.001) after a 3-week washout. At 6 weeks after MPA, the PaCO2 had returned to baseline. The mean changes in PaO2 (+6.0+/-18.0 mm Hg on MPA and +3.8+/-22.5 mm Hg after a 3-week washout) were not significant. The PaO2/PaCO2 ratio increased, and bicarbonate and base excess decreased (p < 0.001) on MPA but not during washout. The systolic BP did not change on MPA but decreased on average 14.8+/-15.0 mm Hg (p = 0.016) after a 3-week washout. The diastolic BP remained unchanged. CONCLUSIONS: Our results suggest that postmenopausal women with chronic respiratory insufficiency consistently improve on MPA at a dose of 60 mg daily for 14 days. Lower PaCO2 is sustained for at least 3 weeks after cessation of MPA. The sustained effects in gas exchange and favorable after-effects in BP warrant further studies into the therapeutic efficacy and possible benefits of MPA pulse therapy. PMID- 10378553 TI - The acute effects of oxygen and carbon dioxide on renal vascular resistance in patients with an acute exacerbation of COPD. AB - OBJECTIVE: Changes in renal hemodynamics occur in patients with severe COPD, especially during an acute exacerbation. Renal hemodynamics are affected by changes in oxygen and carbon dioxide levels, but these changes have not been well defined, particularly in the acute clinical situation. We wished to determine whether oxygen or carbon dioxide levels have the predominant effect on renal hemodynamics in patients with an acute exacerbation of COPD. DESIGN: Fourteen patients with an acute exacerbation of COPD and a PaO2 < 64 mm Hg were studied. Initially, the patients breathed room air (hypoxemia). Then their arterial oxygen saturation was raised to approximately 95% (normoxemia) and then to 98 to 99% (hyperoxemia). Finally, 1 L/min of carbon dioxide was added to the circuit (hyperoxemic hypercapnia). Using duplex ultrasonography, the pulsatility index (PI) of an intrarenal artery was measured after 10 min at each level of oxygenation. The PI is an index of distal renovascular resistance. RESULTS: The PI fell significantly from room-air values on inducing hyperoxemia (p < 0.05). This suggests decreased renovascular resistance and increased renal blood flow. When hyperoxemic hypercapnia was induced, the PI rose significantly from the hyperoxemia level (p < 0.001). CONCLUSIONS: In hypoxemic patients, renovascular resistance decreased when hyperoxemia was induced. This fall in renovascular resistance was reversed with the addition of carbon dioxide. This suggests that acute changes in carbon dioxide levels might have a more dominant role than oxygen levels in determining renovascular resistance. PMID- 10378554 TI - Long-term follow-up after coronary artery bypass grafting reoperation. AB - BACKGROUND: Coronary artery bypass grafting (CABG) reoperation is being performed with increasing frequency. OBJECTIVE: To determine the clinical outcome and the long-term results of a second CABG. SETTING: An 1100-bed urban university affiliated hospital. DESIGN: Retrieval of data on selected parameters from medical records before surgery and prospective follow-up afterwards. PATIENTS AND METHODS: We studied the outcomes of 498 consecutive patients who underwent CABG reoperation in our institution from January 1978 to December 1989 and who were followed postoperatively. Their perioperative mortality, morbidity, and long-term follow-up results were re-evaluated. The end points of the study were December 1997, 15 years of follow-up, or the patient's death. RESULTS: The perioperative mortality rate was 3%. The cumulative survival rates were 90.1%, 74%, and 63.4% at the 5-year, 10-year, and 15-year follow-ups, respectively. The cardiac event free survival rates were 91.5%, 83.4%, and 67.8% at the 5-year, 10-year, and 15 year follow-ups, respectively. The risk factors adversely affecting long-term survival were advanced age, hypertension, and a low left ventricular ejection fraction (LVEF). CONCLUSIONS: The long-term results of cumulative survival and cardiac event-free survival in patients who underwent CABG reoperation are good. Although this reoperation is safe overall, advanced age, hypertension, and a decreased LVEF significantly increase the surgical risk. PMID- 10378555 TI - Determinants of hospital mortality after coronary artery bypass grafting. AB - OBJECTIVES: To examine causes of death and to find predictors of hospital mortality after elective coronary artery bypass graft (CABG) surgery. DESIGN: Case-control study. SETTING: Tertiary teaching hospital. METHODS: We prospectively collected various preoperative, operative, and immediate postoperative variables in a cohort of patients undergoing elective CABG surgery. RESULTS: Of the 2,014 consecutive patients (mean [+/- SD] age of 61.3+/-6.7 years old) undergoing elective CABG over a 2-year period, 27 patients (1.3%) died during their hospitalization. The main causes of death (either isolated or in combination) were cardiogenic shock (n = 13), brain death or stroke (n = 7), septic shock (n = 4), ARDS (n = 2), and pulmonary embolism (n = 1). A univariate statistical analysis revealed factors that significantly correlate with outcome: patient age, preoperative left ventricular ejection fraction, bypass time, aortic cross-clamp time, number of blood units transfused, number of inotropic agents administered in the operating room during the first postoperative day (POD), history of arterial hypertension, intra-aortic balloon pump usage, and perioperative development of shock. A logistic regression analysis showed that the combination of the number of inotropes and the number of blood units administered in the operating room during POD 1 was the most important determinant of outcome, with an overall positive predictive value of 91.7%. CONCLUSIONS: We conclude that the analysis of simple variables enhances our ability to accurately predict hospital mortality in patients undergoing elective CABG surgery. The number of inotropic agents and blood transfusions administered during the immediate postoperative period is the most important independent predictor of hospital mortality. PMID- 10378556 TI - Leukocyte infiltration and secretion of cytokines in pleural drainage fluid after thoracic surgery: impaired cytokine response in malignancy and postoperative complications. AB - STUDY OBJECTIVE: To assess the postoperative course of pleural leukocyte counts and cytokine concentrations in patients with malignant and nonmalignant lung disease who underwent thoracic surgery. PATIENTS AND INTERVENTIONS: A total of 21 patients undergoing thoracic surgery were included in the study. Twelve patients had a malignant disease, and 9 had a nonmalignant disease. Six patients underwent video-assisted thoracoscopy and 15 underwent thoracotomy. Pleural drainage fluid from the chest tubes was collected postoperatively at Oh, 3h, 6h, 12h, 24h, 48h, 72h, and 96 h. The same schedule, as well as one additional preoperative sample, was applied for blood collections. RESULTS: A trend toward lower concentrations of tumor necrosis factor-alpha (TNF-alpha), granulocytemacrophage colony stimulating factor, and interleukin-10 was observed in patients with malignant disease compared to those without malignancy. These differences achieved significance for TNF-alpha in the drainage fluid of those patients with nonmalignant disease who had undergone formal thoracotomy. Patients with malignant disease showed significantly lower macrophage fractions in drainage fluid and lymphocyte fractions in serum. All patients with complications had malignant disease and showed the lowest cytokine concentrations, as well as the lowest fractions of both macrophages in drainage fluid and lymphocytes in serum. CONCLUSION: The data suggest that malignancy may lead to impairment of the wound healing process via modification of the inflammatory cell infiltrate and locally released cytokines. PMID- 10378557 TI - Induced sputum in patients with newly diagnosed sarcoidosis: comparison with bronchial wash and BAL. AB - OBJECTIVES: Sarcoidosis is characterized by a diffuse alveolar inflammatory process, although bronchial airways are often involved. This study compares the cellular profiles of induced sputum (IS), bronchial washing (BW), and BAL in newly diagnosed sarcoidosis patients to those in control subjects, and examines whether inflammatory cell counts from IS are correlated with inflammatory cell counts from BW and BAL in sarcoidosis patients. PATIENTS AND MEASUREMENTS: We recruited 15 untreated patients with stage I and II pulmonary sarcoidosis and 12 healthy volunteers. Sputum was induced with hypertonic saline solution in all individuals. Bronchoscopy was performed on a different occasion in all patients and in five control subjects. RESULTS: Mean lymphocyte counts in IS, BW, and BAL fluid from sarcoidosis patients were significantly higher than in control subjects (9.4% vs 3.8%, p < 0.05; 12.6% vs 3.9%, p < 0.05; 24.1% vs 2.6%, p < 0.05, respectively). Moreover, total cell count and percentage of epithelial cells in IS were significantly higher in sarcoidosis patients than in control subjects (p < 0.01 and p < 0.05, respectively). In sarcoidosis patients, comparison between different samples showed significantly higher percentages of macrophages in BW and BAL than in IS (p < 0.05 and p < 0.01, respectively), whereas the percentage of neutrophils was higher in IS compared with BW and BAL (p < 0.01 and p < 0.001, respectively). Finally, the percentage of lymphocytes in IS was significantly lower than that in BAL (p < 0.05) but not that in BW. CONCLUSIONS: We demonstrated that, compared with IS in healthy control subjects, IS in untreated pulmonary sarcoidosis patients contains more total cells, lymphocytes, and epithelial cells. Although the relative proportion of inflammatory cells in the three samples differed, lymphocyte counts in IS were high. This finding suggests that IS could be used as a valuable alternative to more conventional invasive techniques in clinical assessment of pulmonary sarcoidosis patients. PMID- 10378558 TI - Efficacy and safety of danaparoid sodium (ORG 10172) in critically ill patients with heparin-associated thrombocytopenia. AB - OBJECTIVE: To evaluate the effectiveness and the safety of danaparoid sodium in the treatment of critically ill patients with standard unfractionated heparin induced thrombocytopenia (HIT) or low-molecular-weight HIT. SETTING: University hospital. PATIENTS AND METHODS: Retrospective analysis of 42 consecutive critically ill patients who were admitted for HIT between October 1992 and February 1997 and were treated either with therapeutic or prophylactic doses of danaparoid sodium. RESULTS: Among the 26 patients treated with therapeutic doses, neither new thrombotic complications nor thrombosis extension was clinically suspected. Two deaths were directly related to lower limb acute arterial thrombosis associated with HIT. Two major hemorrhagic complications were observed when aspirin in addition to danaparoid sodium was administered. When danaparoid sodium was used in prophylactic doses (20 courses of treatment) to prevent either postsurgical or medical thrombotic complications, no thrombotic event was observed. No death related to HIT or danaparoid sodium treatment was observed. One aggravation of a postsurgical cerebral lesion was observed. During danaparoid sodium treatment, a persistence or a recurrence of thrombocytopenia was observed in 6.5% of patients without thrombotic complications. CONCLUSION: Danaparoid sodium appears to be an efficient and safe treatment in critically ill patients with HIT. The concomitant use of aspirin in addition to danaparoid sodium seems to represent an important additional hemorrhagic risk that should be avoided in patient management. PMID- 10378559 TI - Rest-redistribution 201-Tl single-photon emission CT imaging for determination of myocardial viability: relationship among viability, mode of therapy, and long term prognosis. AB - BACKGROUND: The diagnosis of viable myocardium in the setting of ischemic left ventricular systolic dysfunction might indicate which patients have the greatest prognostic benefit from myocardial revascularization. Single-photon emission CT (SPECT) thallium-201 (201Tl) scintigraphy for the detection of viable myocardium is widely available in the community, but outcome data using this imaging modality are limited. METHODS: Thirty-seven patients (mean [+/- SD] age, 62+/-12 years) with ischemic left ventricular dysfunction (mean ejection fraction, 30+/ 9%) initially referred for rest-redistribution SPECT thallium scintigraphy were evaluated 29+/-19 months after coronary bypass surgery (n = 15) or medical therapy alone (n = 22). The relationship among myocardial viability, mode of therapy, and long-term prognosis was evaluated. RESULTS: Significant myocardial viability (defined as a viability index [VI] of > 0.5) was present in 19 patients. Among patients with a VI > 0.5, the 48-month actuarial event-free survival was 89+/-10% for patients undergoing surgical revascularization, compared with 0% for the medical treatment subgroup (p = 0.005). In contrast, patients in the low-viability subgroup tended to have intermediate event-free survival rates, which were not statistically different for patients receiving either surgical (62+/-21%) or medical therapy (50+/-14%; p = 0.55). CONCLUSIONS: Survival is significantly more favorable for surgically revascularized patients with ischemic left ventricular dysfunction and myocardial viability as detected by SPECT 201Tl scintigraphy. PMID- 10378560 TI - Myopathy following mechanical ventilation for acute severe asthma: the role of muscle relaxants and corticosteroids. AB - BACKGROUND: Acute myopathy following mechanical ventilation for near-fatal asthma (NFA) has been described recently, and some researchers have suggested that this complication is related to the use of neuromuscular blocking agents (NMBAs) and corticosteroids (CSs). OBJECTIVES: To determine the incidence of acute myopathy in a group of patients and to examine the most important predictors of its development. DESIGN AND METHODS: A retrospective cohort study over a 10-year period (1985 to 1995) of all asthma patients who received mechanical ventilation at two centers in Vancouver (designated center 1 and center 2). RESULTS: In center 1, there were 58 patients who had 64 episodes of NFA, and in center 2, there were 28 patients who had 30 episodes. NMBAs were used in 30 of 86 admissions for acute severe asthma (35%). The mean (+/- SD) duration of muscle paralysis was 3.1+/-2.3 days. A total of 9 patients (10.4%) developed significant myopathy. The incidence of myopathy was 9 of 30 (30%) among patients who received NMBAs. In a multiple logistic regression model, the development of myopathy was only significantly associated with the duration of muscle relaxation. The odds ratio for the development of myopathy increased by 2.1 (95% confidence interval, 1.4 to 3.2) with each additional day of muscle relaxation. The dose and the type of the CS were not significantly associated with the myopathy in the multiple logistic regression analysis. CONCLUSION: Our study showed that there is a high incidence of acute myopathy when NMBAs are used for NFA. The incidence of myopathy increases with each additional day of muscle relaxation. PMID- 10378562 TI - Diagnosis of nosocomial pneumonia in cancer patients undergoing mechanical ventilation: a prospective comparison of the plugged telescoping catheter with the protected specimen brush. AB - STUDY OBJECTIVES: Quantitative culture of protected samples of lower respiratory tract secretions obtained by a fiberoptic protected specimen brush (PSB) is widely accepted for the diagnosis of ventilator-associated pneumonia (VAP), but this diagnostic procedure is time consuming, expensive, and may give rise to iatrogenic complications, especially in cancer patients who often present with thrombocytopenia. The plugged telescoping catheter (PTC) could be a satisfactory alternative to the PSB in this setting. The aim of the present study was to evaluate the interest of the PTC to diagnose VAP in ventilated cancer patients. DESIGN: A prospective observational study. SETTING: A 15-bed medical-surgical ICU in a comprehensive cancer center. PATIENTS AND INTERVENTIONS: Over a 9-month period, 42 patients suspected of having bacterial VAP during mechanical ventilation underwent 69 bronchial samplings: a blinded PTC and a fiberoptic PSB were performed successively in each case. A positive culture for both sampling procedures was defined as the recovery of > or = 10(3) cfu/mL of at least one potential pathogen. The PSB result was taken as the reference standard. MEASUREMENTS AND RESULTS: The overall agreement between the techniques was 87% (60/69). PTC had a sensitivity of 67%, a specificity of 93%, a positive predictive value of 71%, and a negative predictive value of 91%. CONCLUSIONS: We conclude that the accuracy of the blinded PTC compares well with that of the PSB for the diagnosis of VAP in cancer patients. The sensitivity of the PTC observed herein, which is slightly lower than that described in previous studies, may be due to the blinded nature of the method: the indications for initial or secondary coupling with a directed sampling method in patients with suspicion of localized pneumonia remain to be determined. PMID- 10378561 TI - Decreasing catheter colonization through the use of an antiseptic-impregnated catheter: a continuous quality improvement project. AB - STUDY OBJECTIVES: To evaluate the use of an antiseptic-impregnated (chlorhexidine and silver sulfadiazine) catheter for the prevention of catheter colonization and catheter-related bloodstream infection (CR-BSI). Then, based on these findings, to implement changes in hospital policy and to assess their effect on a hospital service. DESIGN: Prospective, randomized, controlled (phase I); prospective, concurrent data collection (phase II). SETTING: Tertiary referral hospital with level 1 trauma center. PATIENTS: Patients > 12 years of age with central venous catheters placed while they were in the emergency room, neurotrauma ICU, or medical/surgical ICU from May through December, 1995 (phase I). All patients > 12 years of age on the trauma service admitted from November 16, 1996, through November 15, 1997 (phase II). INTERVENTIONS: Randomization table determined whether the patient would receive an antiseptic-impregnated catheter (AIC) or nonimpregnated catheter (NIC) (phase I). All removed or exchanged catheters were sent for semiquantitative culture. In phase II, only AICs were used; "length of time" and "fever" were discouraged as reasons for catheter exchange or removal; and only the tip was sent for culture. MEASUREMENTS AND RESULTS: In phase I, there were 139 catheters placed in 60 patients in the NIC group and 98 catheters placed in 55 patients in the AIC group. Two catheters (2.0/100 catheters) in the AIC group were found to be colonized, compared with 25 (18.0/100 catheters) in the NIC group (p = 0.001). The catheter colonization rates were 2.27/1,000 catheter days (AIC) and 24.68/1,000 catheter days (NIC) (p < 0.001), while the CR BSI rates were 1.14/1,000 catheter days (AIC) and 3.9.5/1,000 catheter days (NIC) (p = 0.31). The reason for each catheter removal/exchange was noted, and only "positive blood culture" was statistically significant overall. The tip segment was found to be positive more often than the intracutaneous segment. In phase II, there were 213 AICs placed in 101 patients. The colonization rate was 3.8/100 catheters (4.52/1,000 catheter days), and CR-BSI rate was 1.0/100 catheters (0.6/1,000 catheter days). The colonization rate for catheters left in place remained low for catheters left in place < 14 days (1.6/100 catheters). Only 11% of catheters were exchanged/removed for reason of "fever," as compared with 23% in phase I. CONCLUSIONS: AICs significantly reduce the rate of central venous catheter colonization. In addition, the apparent protective effects of the catheter over time permit less frequent exchanges or removals of the catheters, decreasing both patient risk and hospital cost. PMID- 10378563 TI - Bedside evaluation of efficient airway humidification during mechanical ventilation of the critically ill. AB - STUDY OBJECTIVE: To determine the correlation between simple rating of condensation seen in the flex-tube connecting the heating and humidifying device used with the endotracheal tube and hygrometric parameters (absolute and relative humidity and tracheal temperature) measured by psychrometry. DESIGN: Prospective randomized clinical trial. SETTING: Medical ICU of Louis Mourier Hospital, Colombes, France, a university-affiliated teaching hospital. PATIENTS: Forty-five consecutive mechanically ventilated critically ill patients. INTERVENTIONS: Patients undergoing mechanical ventilation were randomly assigned to receive humidification with one of the four heat and moisture exchangers (HMEs) tested or with a conventional heated humidifier. MEASUREMENTS: The hygrometric performances of four HMEs (BB2215, BB50, and BB100 from Pall Biomedical, Saint-Germaine-en Laye, France; and Hygrobac-Dar from Mallinckrodt, Mirandola, Italy) and a heated humidifier (Fisher & Paykel; Auckland, New Zealand) were studied after 3 h and also after 48 h of use for the Hygrobac-Dar and correlated to a clinical visual inspection rating the amount of condensation in the flex-tube of the endotracheal tube. RESULTS: A total of 95 measurements in 45 patients were performed. The best hygrometric parameters were obtained with the heated humidifier (p < 0.001). The Hygrobac-Dar yielded significantly higher values for both humidities and tracheal temperature than the other three HMEs (p < 0.001). The performance of Hygrobac Dar was unchanged after 48 h of use. There was a significant correlation between the condensation seen in the flex-tube and the hygrometric parameters measured by psychrometry (absolute humidity, rho = 0.7; relative humidity, rho = 0.7; tracheal temperature, rho = 0.5, p < 0.0001). CONCLUSION: In mechanically ventilated ICU patients, visual evaluation of the condensation in the flex-tube provides an estimation of the heating and humidifying efficacy of the heating and humidifying device used, thus allowing the clinician bedside monitoring of airway humidification. PMID- 10378564 TI - A comparison of bronchodilator therapy delivered by nebulization and metered-dose inhaler in mechanically ventilated patients. AB - BACKGROUND: The optimal method of delivering bronchodilators in mechanically ventilated patients is unclear. The purpose of this study was to compare the pulmonary bioavailability of albuterol delivered by the nebulizer, the metered dose inhaler (MDI) and spacer, and the right-angle MDI adaptor in ventilated patients using urinary analysis of drug levels. METHODS: Mechanically ventilated patients who had not received a bronchodilator in the previous 48 h and who had normal renal function were randomized to receive the following: (1) five puffs (450 microg) of albuterol delivered by the MDI with a small volume spacer; (2) five puffs of albuterol delivered by the MDI port on a right-angle adaptor; or (3) 2.5 mg albuterol delivered by a nebulizer. Urine was collected 6 h after the administration of the drug, and the amounts of albuterol and its sulfate conjugate were determined in the urine by a chromatographic assay. RESULTS: Thirty patients were studied, 10 in each group: their mean age and serum creatinine level were 62 years and 1.3 mg/dL, respectively. With the MDI and spacer, (mean +/- SD) 169+/-129 microg albuterol (38%) was recovered in the urine; with the nebulizer, 409+/-515 microg albuterol (16%) was recovered in the urine; and with the MDI port on the right-angle adaptor, 41+/-61 microg albuterol (9%) was recovered in the urine (p = 0.02 between groups). The level of albuterol in the urine was below the level of detection in four patients in whom the drug was delivered using the right-angle MDI adaptor. CONCLUSION: The three delivery systems varied markedly in their efficiency of drug delivery to the lung. As previous studies have confirmed, this study has demonstrated that using an MDI and spacer is an efficient method for delivering inhaled bronchodilators to the lung. The pulmonary bioavailability was poor with the right-angle MDI port. This port should not be used to deliver bronchodilators in mechanically ventilated patients. PMID- 10378565 TI - Effect of combined kinetic therapy and percussion therapy on the resolution of atelectasis in critically ill patients. AB - BACKGROUND: Some critically ill patients have difficulty in mobilizing their respiratory secretions. These patients can develop pulmonary atelectasis that may result in hypoxemia. There are some data to show that atelectasis may be prevented by turning a patient from side to side utilizing special beds. STUDY OBJECTIVES: To determine the role of kinetic therapy (KT) combined with mechanical percussion (P) in the resolution of established atelectasis of the lungs and hypoxemia in critically ill, hospitalized patients. (KT was defined as rotation of a patient along the longitudinal axis of > or = 40 degrees to each side continuously.) DESIGN: Prospective and randomized study (2:1 test to control group). PATIENTS: Twenty-four patients with respiratory failure, either mechanically ventilated or spontaneously breathing, who demonstrated segmental, lobar, or unilateral entire lung atelectasis were studied. SETTING: Medical ICU and adult respiratory ward in a county hospital in New York. INTERVENTIONS: Seventeen patients were treated with KT combined with mechanical P using a KT system (Triadyne Kinetic Therapy System; KCI; San Antonio, TX). Seven patients received manual repositioning and manual P every 2 h. Both groups received similar conventional therapy with inhaled bronchodilators and suctioning. RESULTS: Partial or complete resolution of atelectasis was seen in 14 of 17 patients (82.3%) in the test group as compared with 1 of 7 patient (14.3%) in the control group. The median duration to resolution of atelectasis was 4 days in the test group. Bronchoscopy was performed in 3 of 7 patients in the control group, but in none of the patients in the test group. A cost of $720 was incurred per patient for utilizing the specialty beds for a mean duration of 4 days. An improvement in oxygenation index occurred in the test group (change in baseline PaO2/fraction of inspired oxygen from 207.4+/-106.7 mm Hg to 318+/-100.7 mm Hg) at the end of therapy, while the control group showed a reduction over a similar duration of time (181.3+/-96.3 mm Hg to 112+/-21.2 mm Hg). CONCLUSIONS: KT and mechanical P therapy resulted in significantly greater partial or complete resolution of atelectasis as compared with conventional therapy. There was a generalized trend toward statistical significance in the improvement of oxygenation and a reduced need for bronchoscopy in the group receiving KT and P therapy. PMID- 10378566 TI - Transbronchial biopsy in the presence of profound elevation of the international normalized ratio. AB - STUDY OBJECTIVE: To identify a level of coagulopathy, reported as the international normalized ratio (INR), that predicts hemorrhage following transbronchial forceps biopsy (TBBx) in an animal model. DESIGN: Crossover blinded study using Yucatan mini-swine (Sus scrofa). SETTING: Tertiary medical center with a dedicated animal research facility. STUDY DESIGN: A two-stage study. In stage 1, flexible fiberoptic bronchoscopy with TBBx was performed to establish the amount of bleeding in animals with normal coagulation systems. Animals then were administered escalating dosages of warfarin to obtain one of several increased INR levels. The endpoint of stage 1 was defined as the INR that resulted in a blood loss of > or = 100 mL in > or = 50% of the study animals. In stage 2, all the animals were to be anticoagulated to the INR level determined in stage 1. Topical and systemic measures would then be administered in an attempt to decrease postprocedure hemorrhage, and the results were recorded. RESULTS: Eighteen animals were enrolled in the study. Despite INR levels > 10, no animals developed a hemorrhagic complication of the transbronchial forceps biopsy (TBBx). Eleven animals had INRs > 7. Four animal deaths were recorded, with three animal deaths attributed to nonpulmonary hemorrhage, each due to a ruptured ovarian cyst. One death was anesthesia related. Stage 2 of the study was not performed due to the extreme INR levels reached in the animals during stage 1 and to the lack of a procedure-related complication. CONCLUSIONS: Our study suggests that INR elevation does not correlate with an increased risk of bleeding following TBBx in this animal model. PMID- 10378567 TI - Role of ischemic preconditioning on ischemia-reperfusion injury of the lung. AB - STUDY OBJECTIVES: Ischemia-reperfusion injury of the lung frequently occurs after cardiopulmonary bypass, after pulmonary thromboembolectomy, and especially during lung transplantation. The protective effects of preconditioning on the heart, liver, bones, and various other organs have been previously evaluated. In this comparative study, we used isolated guinea pig lungs to show the effects of preconditioning on lung ischemia. METHODS: The lungs (n = 10 in each group) were mounted on a modified Langendorff perfusion apparatus and perfused by Krebs Henseleit solution for 30 min. We applied an ischemic preconditioning (5 min ischemia + 5 min perfusion, two times) in the experimental group. After 3 h of normothermic ischemia, the lungs were reperfused for 30 min. Pulmonary artery pressures and malondialdehyde (MDA) and glutathione (GSH) levels of the tissue and the perfusate were measured before and after the ischemic period and also at the end of reperfusion. Electron microscopic evaluation was done on randomly selected lungs of three animals in each group at the end of the experiment. RESULTS: Both MDA and GSH levels of tissue and perfusate decreased in the experimental group after reperfusion, although the reduction in GSH levels did not reach statistical significance. The increase in pulmonary artery pressure was lower in the preconditioning group after reperfusion. CONCLUSIONS: Our data showed that ischemic preconditioning of the lung may have a protective effect in ischemic-reperfusion injury. PMID- 10378569 TI - Technetium Tc 99m sestamibi myocardial perfusion imaging: current role for evaluation of prognosis. AB - Like 201Tl imaging, technetium Tc 99m sestamibi (MIBI) myocardial imaging can be used with exercise and pharmacologic testing to assess the presence of coronary artery disease. An increasing body of literature indicates that MIBI can also be used to assess risk of future cardiac events such as myocardial infarction or death. This article summarizes the current status of MIBI imaging for evaluating prognosis in patients with known or suspected coronary artery disease. PMID- 10378568 TI - Is beta-adrenergic-mediated airway relaxation of salmeterol antagonized by its solvent xinafoic acid? AB - STUDY OBJECTIVE: Isolated case reports of asthmatic fatalities accompanied by the use of salmeterol have raised the question whether a paradoxical effect of salmeterol or its vehicle on the airways might contribute to these fatalities. We questioned whether salmeterol's solvent, xinafoic acid, has detrimental effects on the tone of airways or on beta-adrenoceptor binding. MATERIALS AND METHODS: Basenji-greyhound dogs were anesthetized and their peripheral airways challenged with xinafoic acid via a wedged bronchoscope technique. Radioligand binding assays were performed in lung membranes prepared from these dogs. RESULTS: In contrast to a methacholine control, xinafoic acid (0.001 to 1.0 mg/mL) aerosolized into the peripheral airways of anesthetized dogs did not increase airway resistance. Xinafoate alone had no significant effect on the specific binding of 125I-cyanopindolol to lung membranes and did not affect the affinity of salmeterol for the beta-adrenoceptor in the absence or presence of xinafoate, respectively (-log concentration that inhibits 50% [IC50] of the high-affinity site, 7.7+/-0.15 and 7.9+/-0.27; -log IC50 of the low-affinity site = 5.6+/-0.44 and 5.3+/-0.28 [n = 4]). CONCLUSION: These findings suggest that xinafoic acid, the solvent for salmeterol, does not have direct airway irritant effects, does not bind to beta-adrenoceptors, and does not impair the binding of salmeterol to beta-adrenoceptors. Thus, xinafoate is unlikely to contribute to the worsening of airway symptoms in asthmatics using salmeterol xinafoate. PMID- 10378570 TI - Thrombolytic therapy of pulmonary embolism: a comprehensive review of current evidence. AB - Pulmonary embolism (PE) is a common disorder that is accompanied by significant morbidity and mortality. Although anticoagulation is the standard treatment for PE, thrombolytic therapy, with its ability to produce rapid clot lysis, has long been considered an attractive alternative. Although many studies have been performed over the past three decades, however, the indications for the use of thrombolytic agents in patients with PE remain controversial. In this article, we review the medical literature and provide evidence-based guidelines for the use of thrombolytic therapy. We will also discuss the practical aspects of PE thrombolysis. PMID- 10378571 TI - Alpha1-adrenergic hypothesis for pulmonary hypertension. AB - Pulmonary hypertension (PH) is a chronic and disabling condition that affects the pulmonary vasculature. Once PH is diagnosed, the prognosis is generally poor with a rapid downhill course. PH management is largely empirical because the underlying pathophysiologic mechanisms that are responsible for the excessive vasoconstrictor and vascular smooth muscle proliferative responses are poorly understood. Based on new information concerning the role of adrenergic receptors in regulating various cellular functions, a new perspective on the genesis of PH has emerged, along with a unifying hypothesis for the role of alpha1-adrenergic receptors present in the pulmonary vasculature as the major contributor to the pathophysiologic changes associated with PH. Adrenergic receptors that are present on vascular smooth muscle cells regulate vascular tone and growth. The alpha1-adrenergic receptors that are present on the small- and medium-sized pulmonary arteries have a unique and greatly enhanced affinity and activity to alpha1-adrenergic agonists. Under physiologic conditions, this helps in regulating vascular tone and maintains an adequate ventilation/perfusion matching. However, the excessive stimulation of alpha1-adrenergic receptors produces not only smooth muscle contraction but also proliferation and growth. The conditions that produce an increase in alpha1-adrenoreceptor gene synthesis, density, and activity (such as hypoxia or changes in vessel wall pressure) or increase the levels of its agonists (such as norepinephrine, appetite suppressants, or cocaine) greatly enhance pulmonary vascular smooth muscle contractile and proliferative responses and lead to the development of PH. An understanding of the role played by these receptors in the pathophysiology of PH would not only help to avoid the use of alpha1-agonists for appetite suppression and other disease states, but also would help in developing new drugs to block these receptors. A further understanding of the alpha1-adrenoreceptor subtypes present in the pulmonary vasculature, the factors that regulate their expression, and their intracellular signaling pathways would help researchers to devise newer therapeutic strategies and, hopefully, to find a cure for this crippling condition. PMID- 10378572 TI - Assessment of hazardous dust exposure by BAL and induced sputum. AB - OBJECTIVES: BAL, an important tool in assessing occupational lung diseases, is unsuitable for screening programs, exposure evaluation, or monitoring hazardous dust because it is an invasive technique. The results of induced sputum (IS) analysis were compared with BAL and evaluated as a possible alternative. METHODS: We compared BAL with IS analysis of 5 workers exposed to asbestos and 14 exposed to silica and hard metals. Pulmonary function tests and BAL were performed by conventional methods. IS induction was performed after a 20-min inhalation of 3.5% saline solution with an ultrasonic nebulizer. Giemsa-stained cytopreparations were differentially counted. T-lymphocyte subsets were analyzed by flow-activated cell sorter, and messenger RNA (mRNA) was transcribed by reverse transcriptase-polymerase chain reaction. Mineralogic particles were analyzed by scanning electron microscopy and polarizing light microscopy and quantified by an analyzer. RESULTS: The percentage of neutrophils was significantly lower in BAL fluid than in IS specimens, whereas no differences were found in the percentage of lymphocytes and subsets profile. Asbestos fibers were found in BAL but not in IS samples from workers exposed to asbestos. Polarizing particles were found in both samples. Similar mineral elements were found in qualitative analysis by scanning electron microscopy. Quantitative studies showed similar size distribution with a small shift toward larger particles in sputum; mRNA showed the same cytokine profile. CONCLUSIONS: A comparison of BAL and IS specimens in the evaluation of the study population yielded similar quantitative and qualitative results. Further research is needed to evaluate the hypothesis that IS, being a noninvasive technique, may be useful in monitoring exposed workers. PMID- 10378573 TI - A 32-year-old pregnant woman with malaise, headache, nausea, right upper quadrant abdominal pain, and facial swelling. PMID- 10378574 TI - Recurrent sinusitis, arthralgias, and progressive dyspnea in a 26-year-old woman. PMID- 10378575 TI - Finger clubbing and a lung mass. PMID- 10378576 TI - Severe systemic inflammatory response syndrome with shock and ARDS resulting from Still's disease: clinical response with high-dose pulse methylprednisolone therapy. AB - Adult-onset Still's disease, the adult variant of the systemic form of juvenile arthritis, is an uncommon systemic inflammatory disorder of unknown etiology characterized by high spiking fevers, neutrophilic leukocytosis, arthritis, and an evanescent rash. There is often a delay in reaching a firm diagnosis. Differential diagnoses include infection, malignancy, and various immunologic disorders. Increased ferritin levels are of particular value in establishing the diagnosis. Clinical response to high-dose corticosteroids may be dramatic. We report a case of a 29-year-old woman who had recently been investigated for fever of unknown origin, and who presented to our hospital with high fever and hypotension. Her condition rapidly deteriorated with the development of ARDS, disseminated intravascular coagulation, and shock. The patient had a markedly elevated serum ferritin concentration of 26,000 ng/mL. High-dose pulse methylprednisolone therapy resulted in a remarkable clinical improvement. Such a severe case of systemic inflammatory response syndrome, masquerading as septic shock, has not been reported previously. PMID- 10378577 TI - Pulmonary manifestations of POEMS syndrome: case report and literature review. AB - Phrenic nerve paresis is an unusual complication of POEMS (polyneuropathy, organomegaly, endocrinopathy, M-protein spike and skin changes) syndrome. In this report, we describe a case of POEMS syndrome in which a 56-year-old woman presented with dyspnea and ventilatory failure due to bilateral phrenic nerve paralysis. To our knowledge, only one other case of phrenic neuropathy in POEMS syndrome has been reported. PMID- 10378578 TI - New ECG changes associated with a tension pneumothorax: a case report. AB - This case report reveals new ECG changes associated with a left tension pneumothorax, specifically, PR-segment elevation in the inferior leads and reciprocal PR-segment depression in the aVR lead. A mechanism of atrial injury and/or ischemia is proposed as the cause, and the ECG changes associated with a left tension pneumothorax are briefly reviewed. PMID- 10378579 TI - Coronary spasm during outpatient fiberoptic laser bronchoscopy. AB - A 63-year-old woman with metastatic breast cancer was referred to our bronchoscopy unit for outpatient laser resection of an endobronchial mass through fiberoptic bronchoscopy. The patient had no history of ischemic heart disease. During the procedure, the patient developed an ST-segment elevation and a complete atrioventricular block. IV nitroglycerin and morphine were effective in treating this episode. In this patient, we were able to demonstrate a focal spasm by postbronchoscopy coronary angiography. PMID- 10378580 TI - Purulent pericarditis with tamponade in a postpartum patient due to group F streptococcus. AB - Bacterial pericarditis with cardiac tamponade is a life-threatening disorder that has been associated with a variety of organisms. There is usually an associated underlying condition or a seeding of the pericardium from an infection elsewhere. We report the development of cardiac tamponade and a subsequent pericardial constriction due to group F streptococcus purulent pericarditis. We believe this to be the first report of a postpartum patient with purulent pericarditis. PMID- 10378581 TI - Improvement of pulmonary hypertension after liver transplantation. AB - Pulmonary hypertension at the end stage of chronic liver disease is not an uncommon situation. This association termed portopulmonary hypertension raises the question of the feasibility of performing orthotopic liver transplantation (OLT). In the case reported herein, there was a favorable outcome after OLT, even though the mean pulmonary artery pressure (MPAP) before transplantation was increased to 45 mm Hg. Before OLT, the cardiac index (CI) was considerably elevated (7.69 L/min/m2), giving evidence of a marked hyperdynamic circulatory state. The CI decreased significantly after OLT (3.38 L/min/m2), and this produced a significant decrease in the MPAP. Our observation suggests that portopulmonary hypertension due to a marked increase in the CI can be managed successfully by OLT. PMID- 10378582 TI - Intraoperative detection of pulmonary thromboemboli with epicardial echocardiography. AB - We report a novel intraoperative use of epicardial echocardiography in detecting and guiding the removal of pulmonary arterial thromboemboli. We describe a patient with a right atrial thrombus that could not be visualized with intraoperative transesophageal echocardiography. Because we suspected acute pulmonary embolization, epicardial echocardiography was used to visualize the right and left pulmonary arteries. Pulmonary thromboemboli were identified, and pulmonary thromboembolectomy was successfully performed. PMID- 10378583 TI - Percutaneous dilation of tracheal stenosis. AB - A high-grade complex tracheal stenosis distal to a tracheostomy tube occurred in a patient with a chronic vegetative state. The stenosis was easily and rapidly dilated at bedside using commercially available percutaneous tracheostomy kit dilators. Following tracheal dilation, a larger tracheostomy tube was inserted, resulting in the splinting of the stenotic area. To my knowledge, this is the first report of such a bedside technique for the dilation of a tracheal stenosis through a tracheostomy. This technique may provide a temporary relief from tracheal obstruction as long as the stenosis is within the reach of the dilator. PMID- 10378584 TI - Is minimally invasive outpatient pneumonectomy the current standard of care for lung cancer? PMID- 10378585 TI - Pulmonary embolism--treatment vs nontreatment. PMID- 10378586 TI - Testing for exercise limitation in obstructive lung disease. PMID- 10378587 TI - Flutter flap. PMID- 10378588 TI - Tuberculin skin testing. PMID- 10378589 TI - Sarcoidosis is a significant cause of bullous emphysema. PMID- 10378590 TI - Catheter fragmentation of pulmonary emboli. PMID- 10378591 TI - Prevention of air leaks after lung surgery. PMID- 10378592 TI - Diverticular disease of the colon: a century-old problem. PMID- 10378593 TI - Suspected hereditary nonpolyposis colorectal cancer: International Collaborative Group on Hereditary Non-Polyposis Colorectal Cancer (ICG-HNPCC) criteria and results of genetic diagnosis. AB - PURPOSE: The aim of this study was to determine the frequency of mutations in the mismatch repair genes in families suspected of having hereditary nonpolyposis colorectal cancer. METHODS: We devised two criteria for families suspected of having hereditary nonpolyposis colorectal cancer (Criteria I and II). Criteria I consist of at least two first-degree relatives affected with colorectal cancer with at least one of the following: development of multiple colorectal tumors including adenomatous polyp, at least one colorectal cancer case diagnosed before the age of 50, and occurrence of a hereditary nonpolyposis colorectal cancer extracolonic cancer (endometrium, urinary tract, small intestine, stomach, hepatobiliary system, or ovary) in family members. Criteria II consist of one colorectal cancer patient with at least one of the following: early age of onset (<40 years); endometrial, urinary tract, or small intestine cancer in the index patient or a sibling (one aged <50 years); and two siblings with other integral hereditary nonpolyposis colorectal cancer extracolonic cancers (one aged <50 years). A questionnaire was mailed to members of the International Collaborative Group on Hereditary Non-Polyposis Colorectal Cancer to determine the mutation detection rate in mismatch repair genes from the families fulfilling these criteria. For comparison the mutation detection rate for families fulfilling the Amsterdam hereditary nonpolyposis colorectal cancer criteria in each institution was also obtained. RESULTS: Data were obtained from eight different institutions (in 7 different countries). In a total of 123 patients from 123 families (67 families fulfilling Criteria I and 56 families fulfilling Criteria II), genetic testing for germline mismatch repair gene variants was performed. Germline mutations of the hMLH1 or hMSH2 genes were identified in 24 families (20 percent). Of these, the mutation detection rate for families fulfilling Criteria I was 28 percent (19/67). The mutation detection rate for families fulfilling Criteria II was 9 percent (5/56). In these eight institutions, the overall mutation detection rate for families fulfilling the Amsterdam hereditary nonpolyposis colorectal cancer criteria was 50 percent (77/154). CONCLUSION: The Criteria I for suspected hereditary nonpolyposis colorectal cancer have the advantages that they can be applied to nuclear families and they can include extracolonic cancers. The results of this study suggest that families fulfilling Criteria I should be offered genetic testing. The relatively low mutation detection rate in those families fulfilling Criteria II suggests that, using current techniques, genetic testing in these families is not practical. PMID- 10378594 TI - Clinical implications of multiple colorectal carcinomas in hereditary nonpolyposis colorectal carcinoma. AB - PURPOSE: An increased incidence of multiple (synchronous and metachronous) colorectal carcinomas has been reported in hereditary nonpolyposis colorectal cancer. This review was undertaken to determine the clinical implications of multiple colorectal carcinomas in hereditary nonpolyposis colorectal cancer. METHODS: A retrospective review of the records of patients in the hereditary nonpolyposis colorectal cancer registry at Roswell Park Cancer Institute who had either synchronous or metachronous colorectal carcinomas was conducted. RESULTS: Twenty-five of 93 patients with documented pathology were found to have multiple colorectal carcinomas. The mean age at diagnosis of the index colorectal carcinoma was 46.7 (range, 28-65) years. There were 7 (7.5 percent) patients with synchronous colorectal carcinomas and 20 (21.5 percent) patients with metachronous colorectal carcinomas. Two of the seven (28.6 percent) patients with synchronous colorectal carcinomas developed a metachronous colorectal carcinoma. In the patients with metachronous colorectal carcinomas, 29 metachronous events were noted: colon (23) and rectum (6). The mean and median time interval for metachronous colorectal carcinomas were 10.9 and 11.8 (range, 1.5-43.8) years, respectively. The mean times to first, second, and third events were 11.7 (range, 1.5-43.5), 7.9 (range, 2.7-18.7), and 12.3 (range, 11.8-12.7) years, respectively. The majority of patients with metachronous colorectal carcinomas did not have stage progression at the diagnosis of the metachronous colorectal carcinomas: 13 patients had lower or same stage at first event, 4 had lower or same stage at second event, and 2 patients had lower stage at third event. Three of 20 patients with metachronous colorectal carcinomas died of their disease. CONCLUSION: Multiple colorectal cancers are common in hereditary nonpolyposis colorectal cancer. Even though stage progression may not be evident at diagnosis of metachronous colorectal cancer, some of these patients will nevertheless die of their disease. PMID- 10378595 TI - Histologic comparison of hereditary nonpolyposis colorectal cancer associated with MSH2 and MLH1 and colorectal cancer from the general population. AB - PURPOSE: Hereditary nonpolyposis colorectal cancer is reported to have special histologic features. This study compares the histologic features of hereditary nonpolyposis colorectal cancer to colorectal cancers from the general population when hereditary nonpolyposis colorectal cancer cases are restricted to families with known MSH2 and MLH1 mutations. METHODS: Thirty-seven cancers from kindreds carrying MSH2 mutations, 27 cancers from kindreds carrying MLH1 mutations, and 37 colorectal cancers from the general population were reviewed by a pathologist blinded to hereditary nonpolyposis colorectal cancer gene status. Tumor grade, growth pattern, Crohn's-like lymphoid reaction, mucin production, extent of disease in the bowel wall, and lymph node status were evaluated. RESULTS: Poor differentiation and Crohn's-like reaction were a feature of 44 and 49 percent of hereditary nonpolyposis colorectal cancer compared with 14 percent (P = 0.002) and 27 percent (P = 0.049) of colorectal cancers from the general population, respectively. There was no difference in growth pattern, mucin production, lymph node involvement, or local extent of disease between hereditary nonpolyposis colorectal cancer and colorectal cancers from the general population. Poor differentiation and lymph node metastases were found in 57 and 49 percent of MSH2 compared with 26 percent (P = 0.002) and 10 percent (P = 0.03) of MLH1-associated cancers, respectively. There was no difference in growth pattern, mucin production, Crohn's-like lymphoid reaction, or local extent of disease between subgroups of hereditary nonpolyposis colorectal cancer. CONCLUSIONS: Poor differentiation and Crohn's-like reaction are more common in hereditary nonpolyposis colorectal cancer than colorectal cancers from general population. Poor differentiation and lymph node metastases are more commonly seen in MSH2 associated cancers than MLH1. Evaluation of the natural history, pathogenesis, and prognosis of colorectal cancer in hereditary nonpolyposis colorectal cancer should include consideration of which mismatch repair genes are mutated and what the specific mutations are. PMID- 10378596 TI - Selection criteria for treatment of rectal cancer with combined external and endocavitary radiation. AB - PURPOSE: The aim of this study was to identify factors predictive of recurrence of rectal tumors treated with combined external and endocavitary radiation. METHODS: Seventy-two patients with rectal cancer were evaluated clinically and with transrectal ultrasound before combined external and endocavitary radiation. Ideal lesions were moderately differentiated, mobile, not ulcerated, <3 cm in diameter, and <12 cm from the anal verge. External radiation (4,500 cGy) was given during five weeks followed by endocavitary radiation (3,000 cGy x 2). Median follow-up was 31 (range, 7-93) months. RESULTS: Pretreatment transrectal ultrasound stages were uT1 (6 patients), uT2 (27 patients), and uT3 (39 patients). Clinical evaluation identified 26 ideal and 46 nonideal tumors. Overall recurrence was 36 percent; mean time to recurrence was 12 months. Ideal lesions recurred less than nonideal (15 vs. 48 percent; P = 0.01). Mobile lesions recurred less than tethered lesions (26 vs. 52 percent; P = 0.048). Transrectal ultrasound stage was predictive of recurrence (0 percent uT1, 22 percent uT2, and 51 percent uT3; P = 0.015). Surgery was possible in 14 of 17 patients with pelvic recurrence only; 11 patients (65 percent) had curative surgery. Distant metastases occurred in nine patients; all had pelvic recurrences, and six died of disease. CONCLUSION: Patients with uT3 or nonideal rectal cancers should not be offered combined external and endocavitary radiation for cure. Transrectal ultrasound stage is the only independent predictor of recurrence. PMID- 10378597 TI - Single-layer continuous colon and rectal anastomosis using monofilament absorbable suture (Maxon): study of 500 cases. AB - PURPOSE: The study purpose was to evaluate the results of continuous, single layer colon and rectal anastomoses using a monofilament absorbable suture material (Maxon). METHODS: Four hundred ninety-two consecutive patients undergoing five hundred colon and rectal anastomoses with the above technique were evaluated for outcome, including anastomotic leakage, stricture, and other complications, by means of chart review. RESULTS: Three patients (0.6 percent) died after surgery and 7 (1.4 percent) developed clinical evidence of anastomotic leakage. Twenty-four percent developed some postoperative complications, most of which were minor. CONCLUSIONS: Continuous, single-layer colorectal anastomosis using monofilament absorbable suture can be performed safely, quickly, and with a favorable cost ratio. Handsewn anastomoses should still be part of the armamentarium of the well-trained surgeon. PMID- 10378598 TI - Determination of factors responsible for the declining incidence of colorectal cancer. AB - INTRODUCTION: After rising for 13 years in the United States, the incidence of colorectal cancer began to fall in 1986 and has continued to drop since then. This report contains an analysis of the pattern of declining colorectal cancer risk by colorectal subsite, race, and gender and a time trend investigation of suspected risk modifiers of colorectal cancer. METHOD: Colorectal cancer incidence data were obtained from the Surveillance, Epidemiology, and End Results Public Use Files from 1973 to 1994. The following exposure variables were assessed, focussing principally on the period 1970 to 1980: dietary fat, fiber, ethanolic beverages, vitamin A, vitamin C, iron, calcium, estrogen, aspirin, energy intake, body mass index, serum cholesterol, body iron stores, cholecystectomy, constipation, cigarette use, physical activity, and colonoscopic polypectomy. Data sources used in these analyses were principally National Health and Nutrition Examination Surveys I, II, and III. RESULTS: After 1985 colorectal cancer incidence declined predominantly in the distal colorectum almost equally in both white males and white females. Some exposures remained unchanged or trended in the wrong direction (dietary fat, calcium, ethanol, energy intake, physical activity, overweight prevalence, and cholecystectomy). Others did not apply equally to both genders (estrogen, aspirin, ethanol, calcium, and cholecystectomy). Others may become significant in the future, such as aspirin, estrogen, or calcium, because their supplementation is now prevalent, but were not in 1970 to 1975. Of all the risk factors or interventions assessed, the one most consistent with the observed pattern of change is increased use of colonoscopic polypectomy. CONCLUSION: The best method to diminish the incidence of colorectal cancer today may be to increase the use of screening colonoscopy and polypectomy. PMID- 10378599 TI - A prospective, randomized study comparing the effect of augmented biofeedback with sensory biofeedback alone on fecal incontinence after obstetric trauma. AB - PURPOSE: This study was designed to compare prospectively the effects of augmented biofeedback with those of sensory biofeedback alone on fecal incontinence and anorectal manometry after obstetric trauma. METHODS: A consecutive cohort of 40 females with impaired fecal continence after obstetric anal sphincter injury were recruited from a dedicated perineal clinic. Patients were randomly assigned to receive either augmented biofeedback or sensory biofeedback alone. All patients were assessed before and after twelve weeks of biofeedback training, using a fecal continence questionnaire and anorectal manometry. RESULTS: Thirty-nine of 40 females recruited completed the study. Continence scores improved in both treatment groups, but the results were better for those who received augmented biofeedback. Anorectal manometry was unchanged by sensory biofeedback, whereas anal resting and squeeze pressures increased with augmented biofeedback. No change in anal vector symmetry was observed in either group. CONCLUSION: Augmented biofeedback training is superior to sensory biofeedback alone in the treatment of impaired fecal continence after obstetric trauma. PMID- 10378600 TI - Pudendal neuropathy and severity of incontinence but not presence of an anal sphincter defect may determine the response to biofeedback therapy in fecal incontinence. AB - PURPOSE: It has been suggested that the severity of fecal incontinence, the presence of pudendal neuropathy, or an external anal sphincter defect does not preclude clinical improvement with biofeedback therapy. A discrepancy, however, is frequently found between subjective improvement and objective results after biofeedback therapy. Our aim was to assess whether severity of fecal incontinence, presence of pudendal neuropathy, or an external anal sphincter defect could influence the results of manometric parameters after biofeedback therapy in patients with fecal incontinence. METHODS: Biofeedback therapy was used to treat 27 patients with fecal incontinence (25 women; mean age, 53; range, 29-74 years), according to a strict protocol. Manometry, pudendal nerve terminal motor latency, and anal ultrasound were performed in all patients before biofeedback therapy. Manometric evaluation of external anal sphincter function was performed after the biofeedback sessions. RESULTS: Eight of 27 patients had a good clinical response to biofeedback, but with no significant difference in their mean amplitude and duration of squeeze pressure before and after biofeedback. There was no relationship between the clinical results of biofeedback therapy and the initial severity of fecal incontinence, pudendal neuropathy, or external sphincter defect. Patients with severe incontinence (incontinence to solids) and pudendal neuropathy failed to improve the amplitude and duration of their maximum voluntary contraction after biofeedback therapy. Patients with mild fecal incontinence (incontinence to flatus, liquids, or both) (P<0.04), without pudendal neuropathy (P<0.02), or with (P<0.05) and without (P<0.05) external sphincter defect improved their external anal sphincter function after biofeedback therapy. CONCLUSION: In patients with fecal incontinence, the severity of symptoms and pudendal neuropathy should be considered as two factors of poor prognosis of favorable manometric results after biofeedback therapy. Improvement, on the other hand, may be expected after biofeedback therapy despite an external anal sphincter defect. PMID- 10378601 TI - Comparative study of transrectal ultrasonography, pelvic computerized tomography, and magnetic resonance imaging in preoperative staging of rectal cancer. AB - PURPOSE: The preoperative assessment of rectal cancer wall invasion and regional lymph node metastasis is essential for the planning of optimal therapy. This study was done to determine the accuracy and clinical usefulness of transrectal ultrasonography, pelvic computed tomography, and magnetic resonance imaging in preoperative staging. METHODS: A total of 89 patients with rectal cancer were examined with transrectal ultrasonography (n = 89), pelvic computed tomography (n = 69), and magnetic resonance imaging with endorectal coil (n = 73). The results obtained by these diagnostic modalities were compared with the histopathologic staging of specimens. RESULTS: In staging depth of invasion, the overall accuracy was 81.1 percent (72/89) by transrectal ultrasonography, 65.2 percent (45/ 69) by computed tomography, and 81 percent (59/73) by magnetic resonance imaging. Overstaging was 10 percent (9/89) by transrectal ultrasonography, 17.4 percent (12/69) by computed tomography, and 11 percent (8/73) by magnetic resonance imaging; and understaging was 8 of 89 (8.9 percent) by transrectal ultrasonography, 12 of 69 (17.4 percent) by computed tomography, and 6 of 73 (8 percent) by magnetic resonance imaging. In staging lymph node metastasis, the overall accuracy rate was 54 of 85 (63.5 percent) in transrectal ultrasonography, 39 of 69 (56.5 percent) in computed tomography, and 46 of 73 (63 percent) in magnetic resonance imaging. The sensitivity was 24 of 45 (53.3 percent) in transrectal ultrasonography, 14 of 25 (56 percent) in computed tomography, and 33 of 42 (78.5 percent) in magnetic resonance imaging; and specificity was 30 of 40 (75.0 percent) in transrectal ultrasonography, 25 of 44 (56.8 percent) in computed tomography, and 13 of 31 (41.9 percent) in magnetic resonance imaging. The accuracy in detection of positive lateral pelvic lymph nodes under magnetic resonance imaging (n = 8) was 12.5 percent. The accuracy in detection of posterior vaginal wall invasion was 100 percent in transrectal ultrasonography (n = 7) and 100 percent in magnetic resonance imaging (n = 3), but 28.5 percent in computed tomography (n = 7). CONCLUSIONS: Both transrectal ultrasonography and magnetic resonance imaging with endorectal coil exhibited similar accuracy and were superior to conventional computed tomography in preoperative assessment of depth of invasion and adjacent organ invasion. Because transrectal ultrasonography is a safer and more cost-effective modality than magnetic resonance imaging, transrectal ultrasonography is an appropriate method for preoperative staging of rectal cancer. Further efforts will be needed to provide a better staging of lymph node involvement. PMID- 10378602 TI - Secondary coloperineal pull-through and double dynamic graciloplasty after Miles resection--feasible, but with a high morbidity. AB - PURPOSE: Until recently, patients who underwent abdominoperineal resections had to cope with a colostomy for the rest of their lives. For some of these patients this colostomy was a terrible burden, physically and mentally. Publications about abdominoperineal pull-through and double dynamic graciloplasty immediately after a Miles resection showed good results. The purpose of this study was to investigate the procedure as a secondary approach after abdominoperineal resections. METHODS: In this study seven patients were evaluated. All had had an abdominoperineal resection and proved to have unbearable problems with their stoma. All had a secondary pull-through and double dynamic graciloplasty, a mean of 8.5 (range, 1.1-34.8) years after the Miles resection. RESULTS: In five patients continence was regained; two were reversed to colostomy because of several complications. Patients who had a successful outcome also suffered from numerous complications, with a total mean hospital stay of 73.8 (range, 27-167) days, a mean of 3.1 (range, 1-6) additional operations, and 1.8 (range, 0-4) readmissions. CONCLUSION: Secondary anorectal reconstruction after abdominoperineal resection is a feasible option, but with a high morbidity. Because of this the procedure was stopped at the beginning of 1997. PMID- 10378603 TI - Effects of nifedipine on anorectal smooth muscle in vitro. AB - INTRODUCTION: Glyceryl trinitrate reduces anal resting pressure and aids the healing of anal fissures. However, some patients develop tachyphylaxis and the fissure fails to heal, suggesting that other agents are needed. This study assesses the effects of nifedipine (a calcium channel antagonist) in modulating resting tone and agonist-induced contractions in human internal anal sphincter (IAS) and rectal circular muscle. METHODS: Smooth muscle strips from the IAS and rectal circular muscle from ten patients undergoing surgical resection were mounted for isometric tension recording in a superfusion organ bath. The effects of noradrenaline and carbachol were assessed in the presence of various perfusates. RESULTS: LAS strips developed tone and spontaneous activity. Noradrenaline produced dose-dependent contractions. In calcium-free Krebs solution, tone and activity were abolished and no contractions were elicited in response to noradrenaline. Nifedipine also abolished tone and spontaneous activity, but contractions to noradrenaline were only slightly attenuated. In contrast, rectal smooth muscle strips developed spontaneous activity but no resting tone and contracted in response to carbachol. In calcium-free Krebs solution, the spontaneous activity and carbachol contractions were abolished. Addition of nifedipine to the perfusate abolished spontaneous activity and greatly reduced contractions. DISCUSSION: These data suggest that spontaneous activity and resting tone are dependent on extracellular calcium and flux across the cells. Agonist-induced contraction in the IAS is attributable mainly to the release of calcium from intracellular stores, whereas rectal circular smooth muscle depends principally on extracellular calcium entering the cell for contraction. The attenuation of contractions in both tissues and the abolition of resting tone in the IAS suggest that nifedipine may be useful in the management of patients with anorectal disorders. PMID- 10378604 TI - Short chain fatty acids are effective in short-term treatment of chronic radiation proctitis: randomized, double-blind, controlled trial. AB - PURPOSE: Short chain fatty acids are the main energy source of colonocytes and their use may be impaired in chronic radiation proctitis. The aim of the present study was to evaluate the therapeutic effect of short chain fatty acid enemas in patients with chronic radiation proctitis. METHODS: A prospective, randomized, double-blind trial comparing short chain fatty acid enemas with placebo was conducted in 19 patients with chronic radiation proctitis. Short chain fatty acid enemas contained 60 mM sodium acetate, 30 mM sodium propionate, and 40 mM sodium butyrate. The treatment period lasted five weeks and patients were followed up for six months. RESULTS: On admission, both groups were similar regarding all parameters evaluated. After five weeks short chain fatty acid-treated patients showed a significant decrease in the number of days with rectal bleeding from the previous week (4.4+/-1.8 to 1.4+/-2.2; P = 0.001) and an improvement of endoscopic score (4.8+/-1.4 to 2.2+/-1.2; P = 0.001). Hemoglobin values were also significantly higher in short chain fatty acid-treated patients (13.1+/-0.9 g/dl vs. 10.7+/-2.1 g/dl; P = 0.02). Mucosal DNA and protein concentrations decreased in both groups but significantly so only in placebo-treated patients (P = 0.05). Changes in histologic parameters were not significant in either group. Although short chain fatty acid-treated patients did not get worse in the next six months, placebo-treated ones gradually improved, and at the end of six months, differences between the two groups were no longer observed. CONCLUSIONS: Short chain fatty acid enemas can accelerate the process of healing in chronic radiation proctitis, but treatment has to be continuous if a complete and sustained clinical, endoscopic, and histologic response is to be obtained. PMID- 10378605 TI - An audit of strictureplasty for small-bowel Crohn's disease. AB - PURPOSE: The aim of this study was to review the long-term outcome of strictureplasty for small-bowel Crohn's disease. METHODS: We reviewed 111 patients who underwent 285 primary strictureplasties (Heineke-Mikulicz, 236; Finney, 49) between 1980 and 1997. RESULTS: Eighty-seven patients (78 percent) had had previous bowel resections. Forty-six patients (41 percent) required synchronous resection for perforating disease (abscess or fistula) or long strictures (>20 cm). The mean number of strictureplasties was three (range, 1 11). There were no operative deaths. Septic complications (fistula or intra abdominal abscess) related to strictureplasty developed in eight patients (7 percent), of whom two required a proximal ileostomy. Abdominal symptoms were relieved in 95 percent of patients. The majority (95 percent) of patients with preoperative weight loss gained weight (median gain, +2 kg; range, -6 to +22.3 kg). After a median follow-up of 107 months, symptomatic recurrence occurred in 60 patients (54 percent). In 11 patients symptomatic recurrence was successfully managed by medical treatment. Forty-nine patients (44 percent) required reoperation for recurrence: strictureplasty alone in 22 patients, resection alone in 19 patients, strictureplasty and resection in 6 patients, and ileostomy alone in 2 patients. Eighteen patients (16 percent) required a third operation. One patient died from a small-bowel carcinoma which developed in the vicinity of a previous strictureplasty. Two of 19 patients with diffuse jejunoileal disease developed short-bowel syndrome, and were receiving longterm parenteral nutrition. Two other patients were taking corticosteroids for recurrent symptoms. All other patients were asymptomatic, receiving neither medical treatment nor nutritional support. CONCLUSIONS: Strictureplasty is a safe and efficacious procedure for small-bowel Crohn's disease in the long-term. PMID- 10378606 TI - Misconceptions about the colonic J-pouch: what the accumulating data show. AB - INTRODUCTION: Since 1986 when the colonic J-pouch-anal anastomosis was first described, it has gained increasing acceptance as the operation of choice for low rectal cancer surgery. However, there still exist several misconceptions about its use, namely anastomotic complications, alterations in anorectal physiology, and functional outcome. METHODS: All relevant articles derived from MEDLINE databases from 1986 to the present were reviewed. Emphasis was placed on reviewing the features that are claimed to make the colonic J-pouch-anal anastomosis superior to a straight anastomosis. RESULTS AND CONCLUSIONS: The colonic J-pouch has a role in ultra-low rectal cancer surgery, with an apparent reduction in the incidence of anastomotic leaks and reduced bowel frequency. Continence is unchanged and defecatory difficulties can be reduced by constructing a small pouch (< or =5 cm). PMID- 10378607 TI - Rectovesical fistula treated by covered self-expanding prosthesis: report of a case. AB - Postoperative rectovesical fistulas require surgical intervention for their treatment. We present a case treated by placement of a silicone self-expanding prosthesis in the rectum and vesical drainage and give technical details of the procedure. PMID- 10378608 TI - Rectal duplication in an adult: unusual cause of a buttock mass. Report of a case. AB - PURPOSE: Duplications of the rectum are extremely rare embryologic events, with almost 70 cases reported in the world literature. We report on a 39-year-old female patient with a duplication of the rectum. METHODS: Physical examination showed a left buttock mass; rectal examination revealed the presence of a painless mass compressing the rectum posterolaterally, confirmed by computerized tomography. RESULTS: The patient was operated on with a abdominal then a sacrococcygeal approach. After a complete excision, the postoperative course was unremarkable. Histology revealed a rectal duplication lined with heterotopic cylindric ciliated epithelium. DISCUSSION: This case shows that the diagnosis of rectal duplication is difficult and can be confused with other types of anorectal pathology. The presence of heterotopic ciliated epithelium has rarely been described. Complete excision of the duplication should be possible in most cases using a transcoccygeal, transanal, or abdominoperineal approach, depending on anatomic considerations. PMID- 10378609 TI - Original technique for small colorectal tumor localization during laparoscopic surgery. AB - INTRODUCTION: Small colonic tumor localization and correct extension of colonic resection is critical in laparoscopic surgery. Currently used techniques are sometimes inconclusive and may carry some morbidity. We describe an original method of small tumor localization during laparoscopic colorectal operations through the use of preoperative clip applications by colonoscopy and intraoperative ultrasound of the colon. METHODS: Eight patients with small colonic lesions necessitating preoperative marking were included into this study. A two-step technique was used. Before the operation two metal clips were endoscopically applied proximally and distally to the lesion site. At surgery an intraoperative ultrasound examination of the colon or rectum surface was performed to localize the clips. Subsequent laparoscopic colon resection was performed. RESULTS: Endoscopic metallic clips were easily applied around the lesion in all cases without complications. No dislodgement of clips was documented. At surgery laparoscopic ultrasound visualized the clips in all cases. The examination took between 5 and 17 minutes with no specific morbidity. The lesions with the surrounding clips were always found in the resected specimen. CONCLUSIONS: Endoscopic metal clipping and intraoperative laparoscopic ultrasound proved to be an easy, safe, and accurate technique in locating small colonic tumors. PMID- 10378610 TI - New method for paracolostomy hernia repair? PMID- 10378611 TI - Internal anal sphincter and endosonography. PMID- 10378612 TI - Fecal seepage in males with paradoxically high resting anal tone may be associated with anismus. PMID- 10378613 TI - Carcinoma arising in anorectal fistulas of Crohn's disease. PMID- 10378614 TI - Incidence of anal fissure in nonselected neurological patients. PMID- 10378615 TI - Assessing the quality of medical subspecialty training programs. PMID- 10378616 TI - Risk factors for heart failure in the elderly: a prospective community-based study. AB - PURPOSE: The risk factors for the development of heart failure are not clearly defined, particularly for older adults. We undertook the current investigation to examine the associations of traditional cardiovascular risk factors, comorbidity, and psychosocial factors with the risk of heart failure during 10 years of follow up in a community-based elderly population. SUBJECTS AND METHODS: We evaluated 1,749 subjects, 65 years of age or older, free of heart failure, myocardial infarction, and angina at baseline, who were participating in the New Haven, Connecticut cohort of the Established Population for Epidemiologic Studies of the Elderly program. Cox proportional hazards regression models were used to determine risk ratios (RR) and 95% confidence intervals (CI). RESULTS: During 13,811 person-years of follow-up, 173 subjects developed incident heart failure, as confirmed by chart review. Five factors were independent predictors of heart failure: male sex (RR = 1.7; CI, 1.3 to 2.4), older age (RR = 1.9; CI, 1.3 to 2.7 for age 75 to 84 years, RR = 3.0; CI, 1.7 to 5.5 for age 85 years and older, compared with < or = 74 years), diabetes (RR = 2.9; CI, 2.0 to 4.3), pulse pressure > or = 70 mm Hg (RR = 2.3; CI, 1.3 to 4.3, compared with <50 mm Hg), and body mass index > or = 28 kg/m2 (RR = 1.6; CI, 1.0 to 2.4, compared with <24 kg/ m2). Myocardial infarction occurred during follow-up in 8% of the cohort and was also an important predictor of heart failure (RR = 21; CI, 15 to 31). CONCLUSIONS: Age and traditional cardiovascular risk factors are associated with the development of heart failure in the elderly. Preventive strategies should focus on the management of diabetes, blood pressure, and weight, in addition to the prevention and management of myocardial infarction. PMID- 10378617 TI - Is upper gastrointestinal endoscopy indicated in asymptomatic patients with a positive fecal occult blood test and negative colonoscopy? AB - PURPOSE: There are no recommendations as to whether endoscopic evaluation of the upper gastrointestinal tract is indicated in asymptomatic patients who have a positive fecal occult blood test and a negative colonoscopy. SUBJECTS AND METHODS: All asymptomatic patients with a positive fecal occult blood test who were referred for diagnostic endoscopy were identified. Patient charts, endoscopy records, and pathology reports were reviewed. RESULTS: During the 5-year study period, 498 asymptomatic patients with a positive fecal occult blood test and negative colonoscopy were evaluated. An upper gastrointestinal source of occult bleeding was detected in 67 patients (13%), with peptic ulcer disease being the most common lesion identified (8%). Four patients were diagnosed with gastric cancer and 1 had esophageal carcinoma. In addition, 74 patients (15%) had lesions that were not considered a source of occult bleeding; these findings prompted a change in management in 56 patients (11%). Anemia was the only variable significantly associated with having a clinically important lesion identified (multivariate odds ratio = 5.0; 95% confidence interval 2.9 to 8.5; P <0.001). CONCLUSIONS: Upper gastrointestinal endoscopy yields important findings in asymptomatic patients with a positive fecal occult blood test and negative colonoscopy. Our data suggest that endoscopic evaluation of the upper gastrointestinal tract should be considered, especially in patients with anemia. PMID- 10378618 TI - A survey of 2,851 patients with hemochromatosis: symptoms and response to treatment. AB - PURPOSE: Hemochromatosis is a genetic disorder of iron absorption that affects 5 per 1,000 persons and is associated with reduced health and quality of life. We sought to determine the type and frequency of symptoms that patients experienced before the diagnosis and the treatments that they received. METHODS: We mailed a questionnaire to 3,562 patients with hemochromatosis who were located using patient advocacy groups, physicians, blood centers, newsletters, and the Internet. RESULTS: Of the 2,851 respondents, 99% were white and 62% were men. Circumstances that led to diagnosis of hemochromatosis included symptoms (35%), an abnormal laboratory test (45%), and diagnosis of a family member with hemochromatosis (20%). The mean (+/- SD) age of symptom onset was 41 +/- 14 years. Symptoms had been present for an average of 10 +/- 10 years before the diagnosis was made. Among the 58% of patients with symptoms, 65% had physician diagnosed arthritis and 52% had liver disease. The most common and troublesome symptoms were extreme fatigue (46%), arthralgia (44%), and loss of libido (26%). Physician instructions to patients included treatment with phlebotomy (90%), testing family members (75%), and avoiding iron supplements (65%). CONCLUSIONS: The diagnosis of hemochromatosis in most patients was delayed. Physician education is needed to increase the detection of patients with the disease and to improve its management. PMID- 10378619 TI - Characteristics of bronchoalveolar lavage fluid in patients with sulfur mustard gas-induced asthma or chronic bronchitis. AB - PURPOSE: To examine the pattern of immunoglobulins and cellular constituents in bronchoalveolar lavage fluid obtained from patients with sulfur mustard gas induced asthma or chronic bronchitis as compared with healthy control subjects. SUBJECTS AND METHODS: We studied two groups of nonsmoking veterans with either bronchial asthma (n = 21) or chronic bronchitis (n = 28) believed to have been caused by sulfur mustard gas exposure and a third group of healthy, nonsmoking, non-sulfur mustard gas exposed controls (n = 17). Bronchoalveolar lavage was performed in all three groups. The cellular constituents, albumin content, and immunoglobulin concentrations were determined. RESULTS: The three groups did not differ in age or in the serum albumin and immunoglobulin concentrations. The volume of bronchoalveolar lavage fluid recovered was approximately 10% less in the patients with asthma and chronic bronchitis (P = 0.008). The proportions of lymphocytes among the bronchoalveolar lavage cells were similar in all three groups, whereas the proportion of eosinophils was greater in lavage fluid from the asthmatic subjects than in either the healthy control subjects or the patients with chronic bronchitis (P = 0.0001). Both the total number of the recovered cells per milliliter of lavage fluid and the proportion of neutrophils were significantly greater in bronchoalveolar lavage from patients with chronic bronchitis than in healthy subjects or in the patients with asthma (all P <0.001). CONCLUSION: The bronchoalveolar lavage cellular constituents of patients with sulfur mustard gas-induced asthma and chronic bronchitis are similar to those that have been observed previously in patients with asthma and chronic bronchitis from other common causes. PMID- 10378620 TI - Failure of early heparin cessation as treatment for heparin-induced thrombocytopenia. AB - PURPOSE: The complications of heparin-induced thrombocytopenia include thrombosis and death. The purpose of the study was to determine whether early heparin cessation can prevent these outcomes. SUBJECTS AND METHODS: We performed a retrospective analysis of consecutive patients with heparin-induced thrombocytopenia diagnosed by platelet aggregometry. Demographic, clinical, and laboratory findings were compared in patients by whether heparin treatment was stopped early (< or = 48 hours) or late (>48 hours) after the onset of thrombocytopenia, as well as between patients with and without thrombosis. Thrombocytopenia was defined as a 50% decline in baseline platelet counts or an absolute platelet count < 100,000/mm3. RESULTS: Of the 113 patients, 38% developed thrombosis and 27% died. One-half of patients had thrombosis diagnosed >24 hours after heparin cessation. No difference in thrombosis or mortality was found in the 40 patients with early heparin cessation [mean (+/-SD) time of cessation 0.7 +/- 0.6 days] compared with the 73 patients with late heparin cessation (5 +/- 3 days). Thrombosis >24 hours after heparin cessation occurred in 61% of the patients in the early group and in 40% of the late group (P = 0.17). In a multivariate analysis, only a lower nadir of the platelet count (percent of baseline) was associated with thrombosis. Neither thrombosis nor the time to heparin cessation were associated with mortality. CONCLUSIONS: Early heparin cessation was not effective in reducing morbid events in patients with heparin-induced thrombocytopenia. Treatment strategies other than heparin cessation alone should be considered in patients with this condition. PMID- 10378621 TI - The effectiveness of a clinical practice guideline for the management of presumed uncomplicated urinary tract infection in women. AB - PURPOSE: Acute uncomplicated urinary tract infection is a common and costly disorder in women. To reduce potentially unnecessary expense and inconvenience, a large staff-model health maintenance organization instituted a telephone-based clinical practice guideline for managing presumed cystitis in which women 18 to 55 years of age who met specific criteria were managed without a clinic visit or laboratory testing. We sought to evaluate the effects of the guideline. SUBJECTS AND METHODS: We performed a population-based, before-and-after study with concurrent control groups at 24 primary care clinics to assess the effect of guideline implementation on resource utilization and on the occurrence of potential adverse outcomes. We measured the proportion of patients with presumed uncomplicated cystitis who had a return office visit for cystitis or sexually transmitted disease or who developed pyelonephritis within 60 days of the initial diagnosis. Relative risks (RR) and 95% confidence intervals (CI) were estimated, adjusting for the effects of clustering within clinics. RESULTS: A total of 3,889 eligible patients with presumed acute uncomplicated cystitis were evaluated. As compared with baseline, guideline implementation significantly decreased the proportion of patients with presumed cystitis who received urinalysis (RR = 0.75; CI, 0.70 to 0.80), urine culture (RR = 0.73; CI, 0.68 to 0.79), and an initial office visit (RR = 0.67; CI, 0.62 to 0.73), while increasing the proportion who received a guideline-recommended antibiotic 2.9-fold (CI, 2.4 to 3.7-fold). In the prospective comparison of the 22 intervention and two control clinics, the guideline decreased the proportion of patients who had urinalyses performed (RR = 0.80; CI, 0.65 to 0.98) and increased the proportion of patients who were prescribed a guideline-recommended antibiotic (RR = 1.53; CI, 1.01 to 2.33). Adverse outcomes did not increase significantly in either comparison. CONCLUSION: Guideline use decreased laboratory utilization and overall costs while maintaining or improving the quality of care for patients who were presumptively treated for acute uncomplicated cystitis. PMID- 10378622 TI - Risk factors associated with symptoms of gastroesophageal reflux. AB - BACKGROUND: Although patients with gastroesophageal reflux are often instructed to change their lifestyle, population-based data on the risk factors for reflux in the United States are lacking. METHODS: We performed a cross-sectional study in an age- and gender-stratified random sample of the population of Olmsted County, Minnesota. Residents aged 25 to 74 years were mailed a valid self-report questionnaire that measured reflux symptoms and potential risk factors. Logistic regression was used to estimate the odds ratios (OR) with 95% confidence intervals (CI) for reflux symptoms (heartburn or acid regurgitation) associated with potential risk factors. RESULTS: Overall, 1,524 (72%) of 2,118 eligible subjects responded. A body mass index >30 kg/m2 (OR = 2.8; CI, 1.7 to 4.5), reporting an immediate family member with heartburn or disease of the esophagus or stomach (OR = 2.6; CI, 1.8 to 3.7), a past history of smoking (OR = 1.6; CI, 1.1 to 2.3), consuming more than seven drinks per week (OR = 1.9; Cl, 1.1 to 3.3), and a higher psychosomatic symptom checklist score (OR per 5 units = 1.4; CI, 1.3 to 1.6) were independently associated with frequent (at least weekly) reflux symptoms. CONCLUSION: Obesity is a strong risk factor for gastroesophageal reflux, although the value of weight reduction remains to be proven. That family history was also a risk factor suggests that there may be a genetic component to the disorder. PMID- 10378623 TI - Idiopathic nonhistaminergic angioedema. AB - PURPOSE: We sought to describe the characteristics of a group of patients with idiopathic nonhistaminergic angioedema and their response to prophylactic treatment with tranexamic acid. METHODS: We identified 25 patients (15 men and 10 women; age at diagnosis 16 to 77 years) who had idiopathic nonurticarial angioedema that was not prevented by histamine-1 (H1) blockers. Known causes of angioedema were excluded by clinical history, physical examination, and diagnostic tests. RESULTS: The median age at the onset of symptoms was 35 years (range 8 to 66). The frequency of attacks was > 12 per year for 16 patients, six to 11 per year for 6 patients, and one to five per year for 3 patients. All patients had cutaneous attacks, 13 (52%) reported swellings of the pharynx or larynx, and 5 (20%) had symptoms consistent with bowel angioedema. Because of the similarities between these patients and patients who are deficient in C1 inhibitor, the 15 patients with severe and frequent attacks were started on prophylactic treatment with the antifibrinolytic agent tranexamic acid, 1 g three times a day orally for 3 months, tapered according to its effectiveness. The symptoms of 11 patients decreased to less than one attack per year, and the remaining 4 patients had partial remissions (less than 4 attacks per year). Fourteen patients are still being treated with tranexamic acid. CONCLUSION: Patients with idiopathic nonhistaminergic angioedema appear to have similar clinical features and response to treatment with tranexamic acid as those who are deficient in C1 inhibitor. This suggests that those two forms of angioedema might have, at least in part, a similar pathogenesis. PMID- 10378624 TI - In pursuit of folly. AB - Despite the agreement of most of the professional organizations that are concerned with medical education and health manpower that there is an increasing physician surplus, little has been done to address this problem. The number of entering first-year residency positions has remained relatively stable during the past several years, with the number of applicants consistently in excess of the number of positions. Nonetheless, national or state incentives to decrease the number of residency positions have been criticized frequently. The reasons for the physician surplus, the resistance to adjusting the size of residency training efforts, and the feasibility of existing solutions to balance physician supply with demand are critically reviewed. PMID- 10378625 TI - Outpatient treatment of deep vein thrombosis: translating clinical trials into practice. AB - To develop a rational approach to outpatient management, we review the pharmacologic properties of low-molecular-weight heparins and their efficacy in clinical trials of deep vein thrombosis treatment. Low-molecular-weight heparins have better bioavailability and more predictable anticoagulant activity than standard heparin and thus can be administered without routine laboratory monitoring. Randomized trials comparing subcutaneous low-molecular-weight heparin administered primarily at home with inpatient intravenous standard heparin have established the safety and efficacy of outpatient treatment of selected patients. However, many patients were excluded from these studies. The benefits demonstrated in carefully controlled clinical trials of outpatient treatment of deep vein thrombosis required a complex multidisciplinary organization of medical care that is not readily achievable in routine practice. A structured protocol is necessary to ensure that patient care is optimal. The essential components of an outpatient program include appropriate patient selection, adequate patient education, daily follow-up during therapy with low-molecular-weight heparin, and easy access to health-care professionals. PMID- 10378626 TI - Acute diarrhea: a practical review. AB - This review provides a practical, simple, and logical approach to the diagnosis and management of patients with acute infectious diarrhea, one of the most common diagnoses in clinical practice. Diarrhea in the immunocompromised host, traveler's diarrhea, and diarrhea in the hospitalized patient are also discussed. Most episodes of acute diarrhea are self-limited, and investigations should be performed only if the results will influence management and outcome. After an adequate history and physical examination, the clinician should be able to classify the acute diarrheal illness, assess the severity, and determine whether investigations are needed. Most patients do not require specific therapy. Therapy should mainly be directed at preventing dehydration. Various home remedies frequently suffice in mild, self-limited diarrhea. However, in large-volume, dehydrating diarrhea, oral rehydration solutions should be used, as they are formulated to stimulate sodium and water absorption. Antidiarrheal agents can be useful in reducing the number of bowel movements and diminishing the magnitude of fluid loss. The most useful agents are opiate derivatives and bismuth subsalicylate. Antibiotic therapy is not required in most patients with acute diarrheal disorders. Guidelines for their use are presented. PMID- 10378627 TI - Roles of leukocyte/endothelial cell adhesion molecules in the pathogenesis of vasculitis. PMID- 10378628 TI - The legacy of war gas. PMID- 10378629 TI - An uncharted country. PMID- 10378630 TI - Pathology of borderline ovarian tumours. PMID- 10378631 TI - Economic evaluation of cancer treatments: a review of the methods. PMID- 10378632 TI - A pilot study of increasing dose intensity of epirubicin and ifosfamide in patients with small cell lung cancer by using recombinant granulocyte colony stimulating factor. AB - The aim of this prospective study was to investigate the feasibility of increasing the dose intensity of chemotherapy in patients with small cell lung cancer (SCLC) by using recombinant human granulocyte colony-stimulating factor (r metHuG-CSF). Seventeen previously untreated patients (11 male, 6 female) were treated with ifosfamide (5.0 g/m2) and epirubicin (80 mg/m2) in two successive cohorts. Eight patients received chemotherapy every 2 weeks and r-metHuG-CSF 5 microg/kg given subcutaneously daily for 10 days (cohort A), and nine patients received chemotherapy at 10-day intervals with r-metHuG-CSF 5 microg/kg subcutaneously given daily for 7 days (cohort B). The relative dose intensity compared with the conventional 3-weekly regimen was 1.5 and 2.1 for cohorts A and B, respectively. Neutropenia-associated fever complicated two and five treatment courses in cohorts A and B, respectively. There were five episodes of grade 3/4 thrombocytopenia. There were no treatment delays in cohort A and one cycle was delayed in cohort B. One patient from each cohort was withdrawn due to toxicity. Grade 3/4 non-haematological toxicity, other than alopecia, was not observed. This study confirms that it is feasible to increase the relative dose intensity of ifosfamide and epirubicin in patients with SCLC to 2.1 by using r-metHuG-CSF and shortening the interval between treatment cycles. PMID- 10378633 TI - Is mammography of value in women with disseminated cancer of unknown origin? AB - Mammography is often requested to try to identify occult primary breast carcinoma in women with metastatic cancer of unknown primary site. This study aimed to investigate whether mammography is of use in these patients in identifying the breast as the origin of the metastatic disease. Thirty-one women with a working diagnosis of metastatic cancer underwent mammography in an attempt to determine the primary site. None of these women had a palpable breast mass. The site of presentation, pathological type of tumour, site of origin, and benefit of mammography and mammography-provoked biopsy were clarified for each patient. The patients were also followed up to determine survival. The commonest sites of presentation were lung (45%), lymph nodes (19%) and abdomen (16%). The primary sites of these cancers were identified with confidence in 27 patients (87%). The commonest known primary tumour sites were lung (45%), breast (16%) and ovary (16%). Abnormal mammograms were detected in four patients (13%), but three of these did not have breast cancer. In one, the site of origin remained indeterminate, as either breast or lung. Five (16%) had a confident diagnosis of breast carcinoma; all of these women had normal mammograms. We conclude that mammography in women presenting with metastatic disease from an unknown primary site is unhelpful and is not recommended. Furthermore, we could not demonstrate its value in women presenting with axillary lymphadenopathy. PMID- 10378634 TI - Accuracy of patient positioning during radiotherapy for bladder and brain tumours. AB - We report the results of a prospective study to quantify inaccuracies in patient set-up during routine radiotherapy for tumours of the brain and bladder, which took place as part of our departmental development. Knowledge of these inaccuracies is required to put into practice the ICRU 50 recommendations regarding clinical target volume and planning target volume. We measured inaccuracies in two dimensions by comparing portal beam films with the simulator check film. Our method used manual measurements, proved to be a very laborious technique, and demonstrated the need for portal imaging. Ninety-five brain and 97 bladder portal films from 30 brain and 30 bladder patients were examined. Displacements greater than 0.5 cm were seen in 13% of brain treatments in the supero-inferior direction and 1% in the anteroposterior direction. With bladder treatments, displacements greater than 0.75 cm were seen in 12% in the supero inferior direction and 5% in the lateral direction. These results are consistent with other previous studies. We identified a very small systematic error in the department, which was not [corrected] considered to be clinically significant. These results are discussed with reference to other similar studies and the ICRU 50 recommendations. PMID- 10378636 TI - The prevalence and associated variables of deep venous thrombosis in patients with advanced cancer. AB - The prevalence of venous thromboembolism (VTE) in cancer patients has been estimated as up to 15% antemortem, and higher (over 50% in pancreatic tumours) postmortem owing to the asymptomatic nature of many episodes of VTE. We investigated the prevalence of deep venous thrombosis (DVT) in a population of 298 hospice inpatients with advanced cancer. They were screened for the presence of DVT using light reflection rheography; 258 (86.6%) patients were evaluable for DVT, which was found in 135 (52%; 95% confidence interval 46-58). Factors associated (multivariate analysis) with the presence of DVT were: poor mobility, reduced serum albumin level and higher serum urea. A DVT risk assessment index was calculated using these variables. The three highest categories all had significant rates of DVT and, although the lowest category had a low rate of DVT, it accounted for less than 10% of all patients tested. DVT is common in patients with advanced cancer. It was found to be significantly associated with the above variables, but a combined index was of limited clinical application. In view of the number of patients identified with DVT, repeated small pulmonary emboli may be responsible for more symptoms than previously recognized in cancer patients. PMID- 10378635 TI - Intravenous itasetron: establishing the effective dose range for the prophylactic control of acute emesis in cancer patients undergoing high-dose cisplatin chemotherapy. AB - Nausea and vomiting induced by chemotherapy are a major cause of distress to patients and reduce compliance with potentially beneficial treatment. Itasetron hydrochloride is a new 5-hydroxytryptamine3 (5-HT3) antagonist with potent antiemetic properties. It is more potent than ondansetron in animal models and in early clinical studies it demonstrates a long half-life and does not undergo hepatic biotransformation before elimination. The aim of this open, uncontrolled study was to establish the effective dose range of itasetron hydrochloride given intravenously (i.v.) to patients due to receive high-dose cisplatin chemotherapy (50-120 mg/m2) for the first time. Thirty-nine patients were enrolled in the trial and received a single i.v. infusion of itasetron hydrochloride at a dose of 17-280 microg/kg body weight before commencing the cisplatin infusion (median dose 90-110 mg/m2). Antiemetic protection was demonstrated by doses in the range of 35-280 microg/kg. The 17 microg/kg dose was not effective. Treatment failure (>5 emetic episodes/24 hours) was reported in only six (16%) of the 38 evaluable patients over all treatment groups. Adverse events were generally mild or moderate and of a similar type and incidence to those of current 5-HT3 antagonists. Physicians' and patients' assessments of efficacy and tolerability of itasetron hydrochloride were similar, the majority rating the treatment as 'good' or 'very good'. In conclusion, itasetron hydrochloride is effective in the dose range 35-280 microg/kg in preventing cisplatin-induced emesis. Taken together with results from a larger dose-finding study, a dose corresponding to 35 microg/kg (equivalent to 2.5 mg itasetron, calculated as free base) has been pursued in Phase III studies with the i.v. formulation. PMID- 10378637 TI - Sentinel node localization and biopsy in breast cancer. AB - Currently, it is routine practice to carry out axillary lymph node dissection at the time of surgical removal of a malignant primary breast tumour. As breast cancer is being diagnosed at an earlier stage in a growing percentage of cases, this procedure is proving unnecessary in a proportion of patients. Axillary lymph node dissection carries a risk of side effects and ideally we should identify the patients who actually require the procedure. The concept of the sentinel node is not new and was developed over twenty years ago. The technique of lymphoscintography was initially used to identify the sentinel node/s in patients with malignant melanoma. A proposed alternative management strategy to routine axillary lymph node dissection in patients with breast cancer is sentinel lymph node localization using lymphoscintography and biopsy of the identified sentinel node. The aim being to accurately predict the disease status of the axilla and consequently determine whether axillary lymph node dissection is indicated. The technique is currently undergoing a multi-centre trial in the UK. During my elective at St Luke's Cancer Centre, Royal Surrey County Hospital, Guildford I was fortunate to observe this procedure being tested, with patients undergoing both lymphoscintography and sentinel node biopsy as well as axillary lymph node dissection. The results seen during my stay were extremely encouraging. PMID- 10378638 TI - Post-transplant lymphoproliferative disorders. AB - Post-transplant lymphoproliferative disorder (PTLD) is a serious complication of organ transplantation, with multiple factors being implicated in its pathogenesis. The clinical presentation of PTLD is atypical, therefore imaging and tissue biopsy are essential for diagnosis. Treatment requires drug combinations that are appropriate to the PTLD. Four patients with PTLD who had undergone orthotopic liver transplantation are documented, and their clinical characteristics and management described. These case histories identify neutropenia to be a problem and show that age does not seem to influence prognosis. PMID- 10378639 TI - Small cell carcinoma of the cervix complicated by pregnancy. AB - We report the results of treatment in a 26-year old patient with stage IB2 small cell carcinoma of the cervix complicated by pregnancy. A pathological complete remission was achieved following sandwich chemotherapy and radiotherapy. The patient remains in clinical remission 14 months after presentation. PMID- 10378640 TI - Avascular necrosis in patients treated with BEP chemotherapy for testicular tumours. AB - Avascular necrosis (AVN) is known to occur after combination chemotherapy for lymphomas and leukaemias that includes high dose corticosteroids, but it has been reported rarely in patients with solid tumours. We describe five recent cases in young men with testicular tumours (three of which were of good prognosis), who had been treated with chemotherapy using dexamethasone as an antiemetic. Dexamethasone is a low cost and effective antiemetic, but it may be responsible for inducing AVN in patients receiving chemotherapy for solid tumours. A prospective survey of the frequency of AVN is justified to quantify the extent of the problem. PMID- 10378641 TI - The management and clinical course of testicular seminoma: 15 years' experience at a single institution. PMID- 10378642 TI - Breast cancer radiotherapy litigation and the clinical oncologist. PMID- 10378643 TI - Breast radiation injury litigation and RAGE. Radiotherapy Action Group Exposure. PMID- 10378644 TI - The Royal College of Radiologists' Clinical Oncology Information Network. British Association of Urological Surgeons. Guidelines on the management of prostate cancer. PMID- 10378645 TI - Ultra low-dose helical CT of the chest: evaluation in clinical cases. AB - OBJECTIVE: To compare the visibility of normal lung structures and pulmonary abnormalities between ultra low-dose helical CT (ULHCT: 6 mA) and low-dose helical CT (LHCT: 50 mA), and to assess the feasibility of ULHCT for lung cancer screening. MATERIALS AND METHODS: The reduction of tube current to 6 mA was achieved by using an alminum filter installed in an X-ray tube. After obtaining informed consent, both ULHCT and LHCT of the whole lung were performed in five volunteers and 51 patients, with scanning parameters of 120 kV, 1 rotation/second, 10 mm collimation, and 20 mm/second table speed, during a single breath hold. Images were reconstructed every 5 mm with a 180-degree interpolation algorithm. Three chest radiologists were independently asked to compare the visibilities of normal lung structures using a four-point grading scale, and the scores were compared between ULHCT and LHCT. Pulmonary abnormalities including nodules (less than 20 mm) and other abnormalities (mass, consolidation) were evaluated using four decision levels (A: ULHCT equal to LHCT, B: inferior to LHCT but acceptable, C: much inferior to LHCT and not acceptable, D: not visible) and compared between ULHCT and LHCT. Visibility was also compared between the apical and non-apical regions. RESULTS: 99% of the normal lung structures were judged to be visible in ULHCT, and 236 of 345 (68%) of the nodules were judged as "A" and 92 (27%) as "B". Even with nodules of less than 5 mm, judgements of "A" and "B" were made in 74% and 23%, respectively. The visibility of nodules on the ULHCT was worse in the apical region than in the non-apical region (p<0.01). Other lung abnormalities were also graded as "A" (61%) or "B" (34%). No nodules or abnormalities were graded as "D". CONCLUSION: ULHCT has the potential to be utilized in lung cancer screening. PMID- 10378646 TI - Fast dynamic MRI of aortic dissection: flow assessment by subsecondal imaging. AB - PURPOSE: The aim of this study was to evaluate fast dynamic MRI, which consists of a subsecondal MRI sequence combined with a rapid contrast medium infusion technique, in examining clinical cases of aortic dissection. MATERIALS AND METHODS: The subjects consisted of 27 patients with aortic dissection. Turbo FLASH imaging of the aorta was conducted to obtain 50 image frames within 40 seconds. RESULTS: Recognition of the intimal flap and differentiation of the true channel from the false channel was easily accomplished in all 15 patients with a patent false channel. The entry site was detected in 13 of 15 patients. Differentiation of slow flow from thrombus in the false channel was possible in all 27 cases, and the relationship between the lesion and the main branches of the aortic arch was identified in 26 of 27 cases. CONCLUSION: Fast dynamic MRI may provide a large amount of information that is useful for the diagnosis of aortic dissection. PMID- 10378647 TI - Clinical evaluation of three-dimensional MR-cholangiopancreatography using three dimensional Fourier transform fast asymmetric spin echo method (3DFT-FASE): usefulness of observation by multi-planar reconstruction. AB - MR-cholangiopancreatography (MRCP) is a new method that is non-invasive and permits volume data collection and three-dimensional expression. With the three dimensional Fourier transform fast asymmetric spin echo (3DFT-FASE) method, a higher spatial resolution can be obtained both in-pain and in slice selecting direction. In this paper, the usefulness of this new technique is investigated in the clinical diagnosis of MIP images and MPR. The study was performed in 10 normal volunteers and 21 patients with abnormalities in the pancreas or bile-duct sustem. The study was done using a 1.5 Tesla super-conductive machine. The MRCP images were interpreted by three radiologists. In most cases good images were obtained. The additional clinical information provided by MPR was remarkably useful in cystic lesions, especially in mucinous cystic neoplasm of the pancreas. Even when the intestine overlapped the pancreas, it was possible to evaluate the pancreatic duct by MPR. Three-dimensional observation and clinically useful diagnosis were possible by utilizing the advantageds of the 3DFT-FASE method appears quite useful in clinical application. PMID- 10378648 TI - Pharmacologically stimulated portal flow measurement by magnetic resonance imaging for assessment of liver function. AB - PURPOSE: To evaluate pharmacologically stimulated portal flow measured by magnetic resonance (MR) imaging for assessment of liver function. MATERIALS AND METHODS: Pharmacologically stimulated portal flow was measured by phase contrast MR imaging in 27 patients when they were undergoing abdominal angiography for liver tumors or gall bladder cancer. The patients included 11 cases of liver cirrhosis and eight of chronic hepatitis. Pharmacological stimulation was done by infusion of 10 microg/Kg of nicardipine hydrochloride into the superior mesenteric artery through an angiographic catheter. We examined the correlation between stimulated or non-stimulated portal flow and biochemical liver function tests. RESULTS: Correlation coefficients and their corresponding p values between non-stimulated portal flow and the indocyanine green residual rate at 15 min after injection (ICG R15), serum albumin (ALB), total bilirubin (TB), cholinesterase (CHE), and hepaplastin test (HP) were--0.414 (0.056), 0.296 (0.134), -0.570 (0.002), 0.289 (0.153), and 0.321 (0.126), respectively, whereas those between stimulated portal flow and ICG R15, ALB, TB, CHE, and HP were- 0.561 (0.007), 0.411 (0.033), -0.509 (0.007), 0.445 (0.023), and 0.494 (0.014), respectively. CONCLUSION: Stimulated portal flow showed better correlations with biochemical liver function tests than non-stimulated portal flow. It is suggested that stimulated portal flow measurement is more useful for the evaluation of liver function than non-stimulated portal flow measurement. PMID- 10378649 TI - Local thrombolytic therapy for superior mesenteric artery embolism: complications and long-term clinical follow-up. AB - PURPOSE: To evaluate the effectiveness and complications of local intraarterial fibrinolysis in selected patients with superior mesenteric artery (SMA) embolism. MATERIALS AND METHODS: Intraarterial thrombolytic therapy was performed for acute SMA embolism in eight patients. Patients were selected for thrombolytic therapy on the basis of absence of peritoneal signs of intestinal necrosis at physical examination and absent findings of bowel infarction by CT. RESULTS: Clinical success was achieved in five patients and technical success in six. Complications included death due to massive shower emboli from the left ventricle in one patient and extravasation in one patient, who required surgery on the following day. Within one month after thrombolytic therapy, one patient each died of myocardial infarction and cerebral infarction due to emboli, and one patient underwent aorto-SMA bypass surgery due to residual stenosis. In the long-term follow-up period (2-7 years), four patients were still alive, with another embolic episode of a lower limb in one patient. One patient died of an unrelated cause without experiencing another embolic episode. CONCLUSION: Intraarterial fibrinolysis may be a therapeutic alternative in the management of SMA embolism in selected patients in whom an early diagnosis can be made. The long-term results depend on the occurrence of another embolic event. PMID- 10378650 TI - Three-dimensional helical CT for treatment planning of breast cancer. AB - PURPOSE: The role of three-dimensional (3D) helical CT in the treatment planning of breast cancer was evaluated. METHODS: Of 36 patients examined, 30 had invasive ductal carcinoma, three had invasive lobular carcinoma, one had DCIS, one had DCIS with minimal invasion, and 1 had Paget's disease. Patients were examined in the supine position. The whole breast was scanned under about 25 seconds of breath-holding using helical CT (Proceed, Yokogawa Medical Systems, or High-speed Advantage, GE Medical Systems). 3D imaging was obtained with computer assistance (Advantage Windows, GE Medical Systems). RESULTS: Linear and/or spotty enhancement on helical CT was considered to suggest DCIS or intraductal spread in the area surrounding the invasive cancer. Of 36 patients, 24 showed linear and/or spotty enhancement on helical CT, and 22 of those 24 patients had DCIS or intraductal spread. In contrast, 12 of 36 patients were considered to have little or no intraductal spread on helical CT, and eight of the 12 patients had little or no intraductal spread on pathological examination. The sensitivity, specificity, and accuracy rates for detecting intraductal spread on MRI were 85%, 80%, and 83%, respectively. CONCLUSIONS: 3D helical CT was considered useful in detecting intraductal spread and planning surgery, however, a larger study using a precise correlation with pathology is necessary. PMID- 10378651 TI - Breast ultrasonography: diagnostic efficacy of a computer-aided diagnostic system using fuzzy inference. AB - We have developed a computer-aided diagnostic (CAD) system using fuzzy inference for breast sonography and have evaluated the performance of the system. Our CAD system is not an automated image processing method, but requires the observer's subjective "scoring." Seven radiologists interpreted 54 breast mass lesions (24 malignant, 30 benign). Six criteria (shape, border, halo, internal echoes, posterior echoes, and edge shadows) were scored using a five-point rating scale. The output was described as a real number from 0.0 to 1.0. For cancer diagnosis, the sensitivity of the radiologists, a six-criteria CAD version, and a four criteria version (excluding posterior echoes and edge shadows) were 63.1%, 82.1%, and 78.0%, respectively. Specificity was 71.0%, 42.9%, and 51.0%, respectively, while accuracy was 67.5%, 60.3%, and 63.0%, respectively. No significant differences in the areas under ROC curve (Az) were found between the observers and CAD system. Several cases assessed as false-negative by observers were correctly diagnosed by the CAD system, and diagnostic efficacy could be improved, especially among novice sonographers. Although the system's high false-positive rate needs to be reduced, it will assist radiologists in generating a level of suspicion for malignancy. PMID- 10378652 TI - An automated image cytometry system for monitoring DNA ploidy and other cell features of radiotherapy and chemotherapy patients. AB - DNA content and distribution in cell nuclei were studied in samples of fine needle aspiration (FNA) from 27 locally advanced breast and head and neck cancers in two going randomized trials that compared accelerated fractionation to standard fractionation radiation in locally advanced breast cancer and head and neck cancer. Two image cytometry methods were compared: a new, fully automated DNA image cytometry system (AIC) and a conventional image cytometry (CIC) system with manual selection, focusing, and segmentation of cells. The results of both techniques were compared on the basis of DNA histogram parameters including DNA index (DI), mean DNA values (MDV), and Auer's DNA histogram patterns. An excellent correlation was achieved between the two imaging techniques in terms of DI (r=0.985, p<0.001) and MDV (r=0.951, p<0.001) as well as between Auer's histogram patterns, where both methods agreed completely. It was concluded in these analyses that the two image cytometry methods were equivalent. However, the AIC offered an advantage by scanning samples in a fully automated way, which represented significant time saving for cytopathologists working with the system, as well as a larger number of cells used in the automated analysis. With the automated image cytometer, 500 relevant cells were collected and analyzed in about 10 minutes, where with the interactive (manual) method, it took typically an hour to collect and analyze only about 250 cells. Seventeen samples were sufficient for flow analysis. Image cytometry and flow cytometry showed good agreement in DI determination; however, three cases reported as diploid by flow cytometry were found to be aneuploid by image cytometry techniques. PMID- 10378653 TI - Fever complicating brachytherapy application in patients with cancer of the cervix. AB - A retrospective study of one decade of intracavitary brachytherapy (ICB) for cancer of the cervix (CC) showed that ICB application was not immediately interrupted in 28 febrile patients. Twenty-eight afebrile women treated in a similar manner served as a control group. Significant differences in cumulative survival, locoregional failure, and complication rates were not observed between the patient groups. These findings suggest that it is not mandatory to promptly discontinue ICB application in the patient with CC when significant body temperature elevation occurs during the administration of ICB. PMID- 10378654 TI - A case of fallopian tube carcinoma: successful preoperative diagnosis with MR imaging. AB - We report a case of fallopian tube carcinoma, successfully diagnosed preoperatively. The patient was a 64-year-old woman. Transvaginal sonography and computed tomography showed a cystic and solid tumor on the left side of the uterus, suggesting ovarian cancer. The tumor was, however, suspected to be a fallopian tube carcinoma on MR imaging. MR images showed a solid mass surrounded by a tube-shaped cystic part. At surgery, a solid and cystic tumor was found in the left fallopian tube. MR imaging may be useful to assist in the diagnosis of fallopian tube carcinoma. PMID- 10378655 TI - CT and MR findings of splenic angiosarcoma. AB - A surgically confirmed primary splenic angiosarcoma is described. Although angiography showed most of the characteristic findings of the present patient, MR imaging provided a useful information as a supplemental tool. The tumor demonstrated low-signal intensity on both T1- and T2-weighted images, which might differ from hemangioma findings. Subacute hemorrhage within the tumor was revealed by MR imaging, suggesting the way in which the tumor grew during a short period of time. This finding was different from reported angiosarcoma MR findings of siderotic nodules within the tumor. After the administration of Gd-DTPA, MR images clearly demonstrated heterogeneous enhancement within the tumor, which corresponded to the pathologic findings of solid parenchyma with necrotic tissues. PMID- 10378656 TI - Radiotherapy for adrenal gland metastasis from lung cancer: report of three cases. AB - Adrenal gland metastasis is often observed during the clinical course of patients with lung cancer. However, treatment of adrenal gland metastasis is seldom considered because of the systemic spread of the disease. Treatment with curative intent is very rare, but palliative treatment may sometimes be considered when symptoms such as flank pain are observed. Three cases of adrenal gland metastasis were reported. Two of them received surgery for lung cancer and developed a sole metastasis of the adrenal gland. Case 1 developed a sole left adrenal gland metastasis with left flank pain 14 months after surgery for large cell carcinoma of the lung. Curative radiotherapy after intra-arterial chemotherapy was given. A good response was obtained, and he has been alive for 2 years and 9 months. Case 2 developed a right adrenal gland metastasis after radiotherapy for brain metastasis, after having received right upper lobectomy because of SCLC. The increase in the size of the right adrenal gland led us to treat the lesion before symptoms developed. Radiotherapy was given on an outpatient basis. Case 3, who was previously treated with chemoradiotherapy for SCLC, developed brain, liver, and bilateral adrenal gland metastasis. Huge adrenal gland metastases displaced the pancreas and caused severe pain with the increase in serum amylase level. Concurrent radiotherapy with systemic chemotherapy was given and remarkable shrinkage of the adrenal gland metastases was obtained together with pain relief. Cases 2 and 3 died after 8 and 4 months, respectively. In some cases, radiotherapy for adrenal gland metastasis is a good palliative therapy even in the advanced stage patients. Radiotherapy can sometimes curatively treat adrenal metastasis from NSCLC, as in our Case 1, in which adrenalectomy appeared difficult at the time of recurrence. PMID- 10378657 TI - A case of follicular thyroid cancer with tracheal stenosis responded to external radiation therapy. AB - We report the case of a 70-year-old man with follicular carcinoma of the thyroid who complained of worsening dyspnea and was successfully treated by external radiation therapy. The total dose given was 61 Gy in 28 fractions. This case suggests that external radiation therapy is effective for the management of differentiated thyroid cancer with critical stenosis of the trachea that is inoperable and difficult to treat with radioiodine. PMID- 10378658 TI - A case of malignant mixed mesodermal tumor (MMMT) of the ovary: MR features before and after chemotherapy. AB - Malignant mixed mesodermal tumor (MMMT) of the ovary is rare. We describe the MR findings of this tumor before and after chemotherapy. Although MR findings were not specific to ovarian MMMT, MR images provided useful information about the effect of chemotherapy. Thus we could select appropriate therapy. PMID- 10378659 TI - A case of Budd-Chiari syndrome successfully treated by transcatheter recanalization of the right hepatic vein and transjugular intrahepatic portosystemic shunt. AB - A 40-year-old man with Budd-Chiari syndrome due to chronic obstruction of the major hepatic veins was successfully treated by interventional radiologic procedures. Initially, partial splenic embolization was performed to improve thrombocytopenia and coagulopathy of blood. Second, recanalization was done by thrombolic therapy. Finally, the TIPS procedure was carried out through the recanalized right hepatic vein. This resulted in great improvement of Budd-Chiari syndrome, a marked decrease in pressure, and disappearance of gastroesophageal varices on endoscopy. PMID- 10378660 TI - Structural features of IgA molecules which contribute to IgA nephropathy. AB - IgA nephropathy (IgAN) is characterised by the mesangial deposition of polymeric IgA1 (pIgA1). pIgA1 production is reduced in the mucosal immune system in IgAN and increased in the marrow; this switch may be secondary to a defect in gammadeltaT cell control of IgA production. However this does not explain the mechanism by which pIgA1 deposits in the mesangium. There is no direct evidence that classical immune complex deposition occurs in IgAN and alternative mechanisms resulting from physicochemical abnormalities of the IgA1 molecule, particular altered glycosylation, have been proposed. IgA1 has a distinctive hinge region which is a site for O-glycosylation. There is reduced terminal galactose on the hinge region O-glycans of circulating IgA1 in IgAN, perhaps due to a defect in B cell beta1,3 galactosyltransferase. A concomitant O-glycan defect in mesangial IgA1 has not yet been proven. Altered hinge O-glycosylation may have substantial impact on the quaternary structure of the IgA1 molecule influencing its capacity to interact with matrix proteins, IgA receptors on mesangial cells and leucocytes, and complement; it may therefore play a key role in the pathogenesis of mesangial deposition of IgA1 and subsequent glomerular injury in IgAN. PMID- 10378662 TI - Methodological approaches to the analysis of IgA1 O-glycosylation in IgA nephropathy. AB - IgA nephropathy (IgAN) is a common form of glomerulonephritis in which IgA1 molecules deposit in the renal mesangium, leading to progressive glomerular inflammatory injury in a significant proportion of patients. The mechanisms underlying the pathogenesis of IgAN remain poorly understood, but altered O glycosylation, a physicochemical abnormality of IgA1 observed in these patients, may be a contributory factor. Although many studies have reported aberrant IgA1 O glycosylation in IgAN, the precise structural nature of the defect remains to be fully characterised, and analysis of IgA1 O-glycans has proved technically challenging. Three main strategies have been employed: lectin binding to the O glycans in situ on the whole IgA1 molecule; mass spectroscopy of isolated O glycosylated glycopeptides; and size/charge separation of free O-glycans released from IgA1. In this review, the basic principles, strengths and weaknesses of each of these methodological approaches are considered, together with a summary of the data obtained from their use. One of the common criticisms of many studies of IgA1 O-glycosylation is the method of IgA1 purification employed, and therefore, this issue is also critically discussed. PMID- 10378661 TI - New approaches to modify glomerular inflammation. AB - Glomerulonephritis remains the leading cause of end-stage renal failure and treatments for these conditions remain non-specific and with significant side effects. The cellular and molecular basis of acute and chronic inflammation is increasingly understood and the work in a number of animal models of nephritis demonstrates the potential of specific molecular interventions. These include preventing the migration of inflammatory cells by inhibiting the effects of chemokines or blocking endothelial/leucocyte adhesion interactions. Within damaged tissue it is possible to decrease the activity of pro-inflammatory cytokines, such as interleukin-1 (IL-1) and tumour necrosis factor (TNF) by using their natural antagonists, namely interleukin-1 receptor antagonist (IL-1ra) and soluble TNF receptors. In addition the behaviour of macrophages can be altered by the effects of anti-inflammatory cytokines including interleukin-4 (IL-4), interleukin-13 (IL-13), interleukin-10 (IL-10), interleukin-6 (IL-6) and transforming growth factor-beta (TGF-beta). By deactivating the inflammatory response of macrophages these cytokines can favour resolution of disease. The ability to use these approaches in clinical practice remains elusive, however the prospect of using gene transfer technology to deliver anti-inflammatory factors directly to the site of inflammation and our increasing understanding of the complexity of the control of inflammation bring such therapies closer. PMID- 10378663 TI - The effect of membrane permeability on ESRD: design of a prospective randomised multicentre trial. AB - The different permeability of high-flux and low-flux dialysis membranes results in different removal capacity, particularly for uremic toxins of middle and large molecular weight. High-flux dialysers have been evaluated in clinical and epidemiological studies for their effect on mortality, morbidity, dialysis related amyloidosis, nutritional status, response to erythropoietin treatment, dialysis tolerance and the preservation of residual renal function. Many of these studies, however, lack a prospective design and randomised treatment allocation, or have too few patients and too short a follow-up. Therefore, this clinical trial was designed to prospectively investigate the long-term effect of membrane permeability on clinical outcome in a larger number of patients. The primary objective is to compare the effect of membrane permeability on mortality of patients on bicarbonate hemodialysis and treated with a minimum dialysis dose. Patients included in the study should have been on hemodialysis for no longer than one month and have serum albumin 4 g/dl or lower. Patients will be randomised to either the experimental or the control group. During the four-week run-in period the treatment parameters will be established in order to achieve the required dialysis dose. During the maintenance period of three to five years regular visits are scheduled to record clinical and laboratory parameters, to measure Kt/V and to adapt the treatment parameters. Altogether a minimum of 660 patients should be enrolled within a two-year recruitment period. PMID- 10378664 TI - Antiphospholipid (aPL) antibodies in end-stage renal disease. AB - Data are few and conflicting about the prevalence and risk factors for antiphospholipid (aPL) antibodies in end-stage renal disease (ESRD). We studied the prevalence, risk factors and clinical manifestations of lupus anticoagulant (LA) and anticardiolipin antibodies (aCL) among ESRD patients (chronic hemodialysis (HD) patients and kidney transplant recipients) and blood donors. LA was assessed in a large cohort (n=180) of patients by the activated partial thromboplastin time (aPTT), dilute Russel's viper venom test (dRVVT) and lupus anticoagulant-sensitive aPTT reagent (PTT-LA). IgM- and IgG-aCL were measured by a solid-phase enzyme-linked immunosorbent assay (ELISA) in 111 patients (61.5%). The prevalence of aPL was low but, it was higher in ESRD than blood donors (8.8% (16/180) vs. 0%, P=0.005); the frequency of aCL was also higher in ESRD than controls (10.8% (12/111) vs. 0%, P=0.002). LA was similar in the study and control groups (2.2% (4/180) vs. 0%, NS). Among HD patients and kidney allograft recipients there was no difference in LA (3.9% (4/101) vs. 0% (0/79), NS) and aCL frequency (18.6% (8/43) vs. 5.9% (4/68), NS). aPL was not associated with sex, age, time on HD, post-transplantation follow-up, ESRD etiology, thrombotic or hemorrhagic events, or type of HD membrane; however, these findings must be interpreted with caution, given the low frequency of aPL. In one HD patient LA activity was associated with multiple thrombosis of the access graft and native veins. In summary, the prevalence of aPL in ESRD is low but nevertheless higher than controls; LA does not appear to be related to membrane bio-incompatibility and it may be linked to vascular thrombosis; the lack of concordance between LA and aCL was apparent. Further studies are needed to clarify the issue of aPL in ESRD. LA testing should be incorporated into the diagnostic evaluation of recurrent thrombotic episodes in patients on HD. PMID- 10378665 TI - Pneumoperitoneum in peritoneal dialysis patients. AB - The prevalence and clinical significance of pneumoperitoneum in peritoneal dialysis (PD) patients is not fully defined in current literature and some reports suggest that unlike in non-PD patients, it is rarely caused by gastrointestinal perforation. We reviewed 403 chest X-ray films of the 118 PD patients following our PD program in 1995-96, in order to define the prevalence of pneumoperitoneum. We found pneumoperitoneum in 3.7% of the X-rays (15/403) from five patients (4.2%). Its causes might have been: faulty bag exchange technique in two cases and extension tube exchange in three. One patient suffered from a simultaneous episode of peritonitis. Our data and the literature review suggest that 0-11% of pneumoperitoneum episodes in PD patients are due to gastrointestinal perforation; the main causes generally are abdominal operations and catheter manipulation. The amount of air is not useful in assessing the cause of pneumoperitoneum, which takes some weeks to disappear. Computed tomography is more sensitive than standard X-ray in diagnosis. PMID- 10378666 TI - Limits of clinical signs and non-invasive techniques in detecting severe acute rejection. AB - We describe a cadaveric renal transplant patient with an early post-transplant period characterized by normal urine output, normal clinical and biological signs, and a slow decrease of serum creatinine; repeated ultrasonography, color doppler ultrasonography and renal angioscintigraphy were normal or consistent with a clinical diagnosis of mild acute tubular necrosis. Nevertheless a core renal biopsy revealed severe steroid-resistant acute rejection with diffuse infiltrates of lymphocytes and initial transmural arteritis. PMID- 10378667 TI - Renal involvement in ANCA-associated systemic vasculitis. PMID- 10378668 TI - Which 5-fluorouracil regimen?--the great debate. AB - 5-Fluorouracil (5-FU) has been available for over 40 years and has been used in a wide variety of different regimens for the treatment of advanced colorectal cancer, a malignancy with a poor prognosis that is common in industrialized countries. However, despite numerous clinical trials in which 5-FU has been used alone and in combination with a variety of modulating agents [chiefly leucovorin (LV)], and has been administered by bolus injection and i.v. infusion, the optimal regimen for the management of advanced colorectal cancer remains unclear, and there are notable national and international variations in clinical practice. The toxicity of 5-FU also remains an obstacle to the achievement of overall clinical benefit in many patients. The introduction of novel chemotherapeutic agents may make it necessary to reassess the place of 5-FU in the treatment of advanced colorectal cancer. This article debates these issues with a review of clinical trials of 5-FU, and concludes that the future lies in the utilization of novel and established agents in combinations that may significantly improve outcomes, rather than in continuing experimentation with various schedules of 5 FU and LV. PMID- 10378669 TI - Weekly 24-h infusion of high-dose 5-flurouracil and leucovorin in patients with advanced gastric cancer. AB - The effect of biochemical modulation of weekly high-dose 5-fluorouracil (5-FU) 24 h infusion by leucovorin (LV) in the treatment of 39 consecutive patients with advanced gastric cancer without prior chemotherapy from October 1996 to August 1997 was examined. There were 21 male and 18 female patients with a median age of 56 years. The regimen consisted of 5-FU 2600 mg/m2 and LV 150 mg administered by 24 h infusion weekly for 6 weeks followed by a 2 week break. The treatment was repeated every 8 weeks until disease progression, patient refusal or unacceptable toxicity. Placement of a central vascular device and a portable external infusion pump was required in all patients and was used for outpatient treatment. The response to treatment was evaluated every 8 weeks. A total of 395 chemotherapy treatments were given with a mean of 10 (2-24). This response rate was: 33% (12 of 36) partial response (PR) rate, 33% (12 of 36) stable disease (SD) and 33% (12 of 36) progressive disease (PD). In general, the toxicity was mild but two toxic deaths occurred, one due to neutropenic sepsis and the other due to hyperammonemia. The median time to progression was 4 months. The overall median survival was 7 months. The survivals of the PR, SD and PD were 12, 8 and 5 months, respectively. This regimen showed a modest activity against gastric cancer with acceptable toxicity. Weekly 24 h infusion of high-dose 5-FU with LV in an outpatient setting for patients with gastric cancer is feasible and deserves further study as a basis for combination therapy. PMID- 10378670 TI - Phase I and pharmacologic study of the arotinoid Ro 40-8757 in combination with cisplatin and etoposide in patients with non-small cell lung cancer. AB - This phase I study was performed to assess the feasibility of combining cisplatin/etoposide (VP-16) with the arotinoid Ro 40-8757 and to determine the dose-limiting toxicity (DLT) of Ro 40-8757 in this combination. Patients with non small cell lung cancer were eligible. Treatment consisted of Ro 40-8757 p.o. day 1-21, cisplatin 100 mg/m2 i.v. on day 2 and VP-16 100 mg/m2 i.v. on day 2-4, repeated every 3 weeks. Eighteen patients were evaluable for toxicity and response. The doses of Ro 40-8757 ranged from 84 mg/m2 once daily to 42 mg/m2 thrice daily (tid). DLT consisting of delayed nausea/vomiting was reached at 42 mg/m2 tid. Consequently, the maximum tolerated dose was set at one dose level below the DLT, i.e. 28 mg/m2 tid. Skin toxicity occurred but was well manageable. Pharmacological analyses showed a small increase in the volume of distribution of cisplatin and VP-16 between the first and third course. However, no relationship with side effects was found. A response was achieved in 50% of patients. The combination of cisplatin/VP-16 with Ro 40-8757 appears to be feasible at a dose schedule of 28 mg/m2 tid. The response rate was at the upper rate of what can be expected with cisplatin and VP-16. PMID- 10378671 TI - Phase II study of a combination of low-dose cisplatin with 13-cis-retinoic acid and interferon-alpha in patients with advanced head and neck squamous cell carcinoma. AB - Preclinical and clinical data have suggested antitumor efficacy in squamous cell carcinoma (SCC) of interferon (IFN)-alpha and 13-cis-retinoic acid (13-c-RA) as single agent with greater activity in combination. Cisplatin was added to potentiate activity. Twenty-three patients with pretreated advanced or metastatic head and neck squamous cell carcinoma were given a combination of IFN-alpha (6 x 10(6) U/day, 84 days s.c.), 13-c-RA (1 mg/kg/day, 84 days) and cisplatin (40 mg/kg/day, day 1, 28 and 56). Seventeen patients had discontinuation of treatment and three patients received overall treatment without dose reduction. Hematological toxicity was more frequent; only three patients experiencing grade 3 or higher extra-hematological toxicity. Four out of 14 evaluable patients were in response, with one in complete pathological response. Median duration of response was 6 months with a 9 month median survival. Association of IFN-alpha, 13-c-RA and cisplatin induces modest but definite antitumor activity with moderate and manageable toxicity. Further studies of different combination modality therapy with chemotherapy and differentiating agents need to be performed in less pretreated patients. PMID- 10378672 TI - A phase II clinical and pharmacological study of oral 9-nitrocamptothecin in patients with refractory epithelial ovarian, tubal or peritoneal cancer. AB - 9-Nitrocamptothecin (9-NC) is a water-insoluble topoisomerase I inhibitor with a broad antitumor activity in animal models. A phase II study was performed in patients with heavily refractory ovarian, tubal or peritoneal cancer (median number of previous chemotherapy regimens > 3) to determine the activity of a daily oral dose of 9-NC. 9-NC dose was 1.5 mg/m2/day for four consecutive days every week. Increments of 0.25 mg/day were authorized in patients without significant side effects. Of 29 evaluable patients, a 7% remission rate was observed. Thirty-four percent of patients had stable disease. The median survival was 8 months. Toxicity was evaluated in 31 patients. Grade 3 or 4 hematologic toxicity consisted of anemia in 10 patients (32%), neutropenia in eight (26%) and thrombocytopenia in three (10%). Grade > or = 2 non-hematologic toxic effects were nausea and vomiting in 26 (84%), diarrhea in 12 (39%), weight loss in seven (22%), chemical cystitis in six (19%) and neutropenic sepsis in six (19%). 9-NC was tolerated for sustained periods of time in some patients (up to 47 weeks). The observed 8-month survival in such a refractory patient population is noteworthy. Further clinical research of prolonged exposure to less toxic analogs of 9-NC is warranted. PMID- 10378673 TI - Continuous delivery of venous 5-fluorouracil and arterial 5-fluorodeoxyuridine for hepatic metastases from colorectal cancer: feasibility and tolerance in a randomized phase II trial comparing flat versus chronomodulated infusion. AB - High-dose chemotherapy combining regional hepatic artery infusion (HAI) of fluorodeoxyuridine (HAI FUDR) and systemic venous infusion of 5-fluorouracil (i.v. 5-FU) was delivered against liver metastases from colorectal cancer. The hypothesis that chronomodulation of delivery rate along the 24 h time scale would improve the tolerable doses of both drugs was tested. Combined HAI FUDR (80 mg/m2/day) and i.v. 5-FU (1200 mg/m2/day) were administered for five consecutive days every 3 weeks, either as a constant rate infusion (schedule A, 27 patients) or as chronotherapy (schedule B, 29 patients). This latter regimen consisted of a sinusoidal modulation of the delivery rate over the 24 h scale with a maximum at 16:00 for FUDR and 4:00 for 5-FU. Intrapatient dose escalation up to the individual maximum tolerated doses (MTD) was planned for both drugs in the absence of any previous grade 3 or 4 toxicity. All patients had metastatic colorectal cancer, with adjuvant or palliative chemotherapy given to six patients (22%) on schedule A and 12 patients on schedule B (41%). Severe stomatitis occurred in 71% of the patients and was dose limiting. No hepatic toxicity was encountered. Dose reductions of 5-FU and/or FUDR were required for 17 of 27 patients on schedule A (63%) as compared to 11 of 29 patients on schedule B (38%), following reaching the individual MTD (p<0.05). Over the first six cycles, patients on schedule B received higher doses (mg/m2/cycle; FUDR: 522 +/- 85 versus 499 +/- 50, p=0.004 and 5-FU: 5393 +/- 962 versus 5136 +/- 963, p=0.009) and higher dose intensities (mg/m2/week; FUDR: 164 +/- 46 versus 151 +/- 52, p=0.018 and 5-FU: 1652 +/- 478 versus 1553 +/- 535, p<0.041) of both drugs than patients on schedule A. As a result the number of courses with doses of 5-FU above 1200 mg/m2/day and/or FUDR above 110 mg/m2/day was larger in group B than in group A (5-FU, A: 67 of 268, 25% versus B: 133 of 321, 41% and FUDR, A: 86 of 268, 32% versus B: 155 of 321, 48%; p<0.001). Objective responses were observed in 13 patients on schedule A (48%) and 11 patients on schedule B (38%). The results support the need for further exploration of chronotherapy of colorectal cancer liver metastases with combined arterial and venous fluoropyrimidine chemotherapy. PMID- 10378674 TI - A phase I trial of topical topitriol (calcitriol, 1,25-dihydroxyvitamin D3) to prevent chemotherapy-induced alopecia. AB - This study evaluated the toxicity and efficacy of topical topitriol (calcitriol, 1,25-dihydroxyvitamin D3) to prevent chemotherapy-induced alopecia (CIA). Patients with breast cancer scheduled to receive FAC chemotherapy (5 fluorouracil, adriamycin and cyclophosphamide) were eligible for the study. Initially, the first six patients were randomized in a double-blind fashion to have received topitriol or placebo with all subsequent patients being treated with topitriol. Topitriol cream (0.0025 or 0.005%; 25 and 50 microg/g concentration) was administered topically twice a day. Three different doses and schedules of administration were evaluated including: 500 and 1000 microg daily for 7 days prior to chemotherapy, and 2000 microg daily for 5 days prior and 5 days post-chemotherapy. Fourteen patients were treated (12 with topitriol and two with placebo) at three different dose levels. All patients developed grade 2 alopecia between day 20 and 30 after chemotherapy, demonstrating the lack of efficacy of topical topitriol on this schedule of administration to prevent CIA. Eight patients exposed to topitriol developed a toxic maculopapular dermatitis in areas exposed to the drug. In conclusion, topical topitriol at the doses and schedules evaluated in this trial was ineffective to prevent CIA and induced a local dermatitis in areas exposed to the drug. PMID- 10378675 TI - A unique paclitaxel-mediated modulation of the catalytic activity of topoisomerase IIalpha. AB - Paclitaxel (Taxol) is known to act by polymerizing and stabilizing microtubules. In spite of a known target, the existence of additional targets is suggested by a poor understanding of the mechanism(s) underlying eventual cell death by paclitaxel and by the drug's high efficacy, as compared to other spindle poisons. Based on the enhanced sensitivity of a mutant DNA double-strand break repair deficient Chinese hamster ovary cell line to paclitaxel as well as to various topoisomerase (Topo) II poisons, it was hypothesized that paclitaxel, in addition to having an effect on microtubules, may also alter the activity of Topo II. This study demonstrates the unique, in vitro effects of paclitaxel on Topo II activity as investigated by monitoring the decatenation of kinetoplast DNA and relaxation of supercoiled plasmid DNA by Topo II. Unlike classical anti-topoisomerase drugs, low concentrations of paclitaxel (0.02-500 nM) stimulated Topo II catalytic activity, while higher concentrations over 5 microM inhibited the activity of Topo II. Furthermore, these effects of paclitaxel appear to be mediated through a direct interaction of paclitaxel with Topo II rather than an interaction with DNA or DNA-Topo II complexes. Collectively, the evidence presented suggests the existence of an atypical interaction between Topo II and paclitaxel that may disrupt the normal functioning of the enzyme. PMID- 10378676 TI - Methotrexate-albumin conjugate causes tumor growth delay in Dunning R3327 HI prostate cancer-bearing rats. AB - Based on the rationale of a preferred albumin uptake by tumors, conjugates comprising of rat serum albumin (RSA) as a drug carrier and of methotrexate (MTX) as chemotherapeutic drug were prepared. For a comparative study of MTX-RSA and MTX we chose a slow growing Dunning R3327 HI prostate cancer model. In a radiopharmacologic study blood kinetics and the tumor and organ distribution pattern of residualizingly labeled MTX-RSA were determined, and were found to be similar to that of residualizingly labeled RSA. The MTD was established for Copenhagen rats at a total four injections of 2 mg/kg MTX or MTX-RSA administered at days 0, 4, 8 and 12. Tumor volume measurements and tumor removal showed a small non-significant growth delay in the MTX treatment group, suggesting MTX resistance for the Dunning R3327 HI prostate carcinoma. In contrast, treatment with MTX-RSA resulted in a significant (50%) growth inhibition of the Dunning R3327 HI tumor. The cellular mechanisms responsible for MTX resistance in Dunning HI tumor cells is not known. The improved therapeutic effects seen during MTX-RSA treatment in this slow growing adenocarcinoma might be a result of prolonged tumor exposure time and an altered cellular uptake by a lysosomal route. MTX albumin conjugates have shown antitumor activity exceeding that of MTX in several tumor xenografts in nude mice, including human prostate cancer. The recently initiated clinical development of MTX-human serum albumin will be continued and cancer of the prostate will be included as a potential target tumor during further clinical phase II testing. PMID- 10378677 TI - Cytotoxic effects of two gamma linoleic salts (lithium gammalinolenate or meglumine gammalinolenate) alone or associated with a nitrosourea: an experimental study on human glioblastoma cell lines. AB - Gamma linoleic acid (GLA) salts may exert a direct antiproliferative activity on tumor cells. The cytotoxicity is linked to the generation of conjugated dienes, peroxyl radicals and superoxide radicals. Lithium gammalinolenate (LiGLA) and meglumine gammalinolenate (MeGLA) have been recently developed for enhancing the water solubility of these compounds. MeGLA or LiGLA (10(-5) to 10(-4) mol/l) and fotemustine (Fote) (2 x 10(-6) to 2 x 10(-4) mol/l) were applied, alone or in combination, for up to 9 days to two human glioblastoma cell lines A172 and U373MG. Fote was applied first followed by LiGLA and/or MeGLA. Cytotoxicity was evaluated by the MTT test, and the effects of drug combinations were analyzed by the isobolographic representation according to the Chou and Talalay method (combination indexes). For both GLA salts, cytotoxicity was manifested after 4 days of cell exposure and with very sharp dose-response curves. Comparison of IC50 values indicated that MeGLA was more active than LiGLA. There was a constant reduction in IC50 values following an increase in exposure time for A172 cells: between 4 and 9 days of cell exposure, IC50 changed from 73 to 46 microM for LiGLA and from 49 to 31 microM for MeGLA (p<0.05). With U373MG cells, there was no influence of exposure duration on IC50 values. Combination index values indicated that association between Fote and GLA salts globally resulted in slightly antagonistic effects. These results may be useful for further development of GLA salts at the clinical level. PMID- 10378678 TI - Cisplatin combined with tiopronin or sodium thiosulfate: cytotoxicity in vitro and antitumor activity in vivo. AB - We have previously reported that the thiol compound tiopronin protects rat kidneys in vitro against the toxic activity of cisplatin. The influence of tiopronin and sodium thiosulfate (STS) on the cytotoxicity of cisplatin has been investigated on P388 leukemic cells in vitro after 3 days. The combination has also been investigated in vivo in BDF1 mice bearing a P388 s.c. tumor. In contrast to STS, tiopronin did not significantly reduce the cytotoxic activity of cisplatin in vitro and nor did it affect the uptake of platinum (cisplatin derived), binding to DNA or the percentage of interstrand cross-links (%ISCL) formation. The co-administration of cisplatin (4 mg/kg) and tiopronin (150 and 300 mg/kg) to BDF1 female mice bearing a s.c. P388 tumor produced a significant reduction in tumor growth similar to that of a single 6 mg/kg dose of cisplatin. Interestingly, pre-incubation in vitro of either tiopronin or STS for 2 h with the species formed from cisplatin by hydrolysis demonstrated their ability in inhibiting the cytotoxicity of these reactive platinum products. These results indicate that tiopronin does not reduce the cytotoxicity of cisplatin in vitro, as STS does. This may be, at least partly, because of a different effect of the two thiol compounds on the cellular uptake and binding of platinum to DNA. Notably, tiopronin substantially reduced tumor growth in mice treated with a non toxic dose of cisplatin (p < or = 0.0277), suggesting some positive influence of this thiol compound on the antitumor properties of cisplatin. The ability of tiopronin to protect in vitro against the cytotoxicity of the aquation products of cisplatin may be related to its nephroprotective effect. PMID- 10378679 TI - Hydroxyurea treatment enhances metastatic cell adhesiveness: a possible chemotherapy-induced increase in metastatic potential? AB - Treatment of tumor cells with certain chemotherapeutic agents, including hydroxyurea, increases their metastatic potential. In the present study the adhesiveness of SW620 colonic metastasis cells treated with hydroxyurea was compared with untreated cells by measuring their ability to remain attached to tissue flask walls during gentle saline washes. Treatment of SW620 cells was found to significantly increase their adhesiveness, suggesting a mechanism by which hydroxyurea may enhance the metastatic potential of tumor cells. PMID- 10378681 TI - Molecular analysis of the combining site of a monoclonal antibody against spermine. AB - The structural basis of the binding of the polyamine spermine to the monoclonal antibody SPM8-2 was studied using computer modelling, ELISA methods and chemical modifications of the binding site residues. Paratope modelling showed that the antibody combining site forms a highly negatively charged cavity mainly shaped by aspartic acid and tyrosine residues which contact the tetra-positively charged spermine molecule by electrostatic interactions and hydrogen bondings. The importance of the electrostatic environment for spermine binding to SPM8-2 is emphasised by the strong dependency on pH and ionic strength. Specific chemical modifications of carboxylate groups and tyrosine residues of the antibody adsorbed to microtiter plates resulted in decreased binding of the N1-biotin spermine conjugate used to monitor the activity of the antibody. These observations are consistent with a key role of aspartate and tyrosine residues in complex formation with spermine. These studies, important to our understanding of antibody-hapten specificity, may also shed light on important motifs responsible for protein-polyamine interactions. PMID- 10378680 TI - PMS2-deficiency diminishes hypermutation of a lambda1 transgene in young but not older mice. AB - The Pms2 gene is involved in DNA mismatch repair in mammalian cells, and has recently been shown to affect hypermutation of mammalian immunoglobulin genes. We have studied hypermutation of a lambda1 transgene in chronically stimulated Peyer's patch B cells of both young and old mice deficient in function of Pms2. In young (3-4 months) mice, somatic hypermutation is fourfold lower in PMS2 deficient mice than in control mice. This difference is statistically significant (P < 0.05). In contrast, in older mice (9 months of age), hypermutation levels are indistinguishable in the Pms2-/- and Pms2+/+ backgrounds. In the older mice, there was no clear difference in the fraction of clones carrying either any mutations or at least two mutations when PMS2-deficient mice were compared with their wild-type littermates. As genomic instability increases with age, this observation is difficult to reconcile with the hypothesis that highly mutated B cells cannot survive in Peyer's patches. Moreover, there were clear differences apparent in the mutation spectra of the Pms2-/- and Pms2+/+ mice. In the PMS2 deficient background, deletion and insertion mutations were found, and there was a significant decrease in the ratio of A mutations to T mutations in comparison with the Pms2+/+ controls. Our data support the hypothesis that PMS2 functions in somatic hypermutation, and are most consistent with the hypothesis that the role of PMS2 is direct rather than indirect. PMID- 10378682 TI - Extracellular processing and presentation of a 69-mer synthetic polypetide to MHC class I-restricted T cells. AB - The classical pathway for MHC class-I-restricted Ag presentation processes cytosolic Ag synthesized in or delivered into the cytosol for binding to MHC class I molecules in the ER. Alternatively, Ag may be processed and bind class I molecules in endocytic compartments or at the cell surface after regurgitation of processed peptides. We show that a 69-mer synthetic polypeptide that carries the optimal 9-mer Kd-restricted epitope from the Plasmodium berghei circumsporozoite protein, PbCS 245-253, is presented to CD8+ T cells after a short incubation (1-2 h) with target cells. The presentation kinetics correlate with the length of the peptides when shorter peptide analogues are used. This presentation is independent of the transporters associated with antigen processing and presentation (TAP), does not require newly synthesized proteins and does not proceed via regurgitation of intracellularly processed peptides. In contrast, it is substantially decreased in the absence of beta2 microglobulin or serum. Taken together, these data suggest that serum components, such as proteases and beta2 microglobulin, allow the processing and loading of exogenous polypeptides onto empty cell surface class I molecules for presentation to CTL. PMID- 10378683 TI - Molecular analysis of the heavy chain of antibodies that recognize the capsular polysaccharide of Neisseria meningitidis in hu-PBMC reconstituted SCID mice and in the immunized human donor. AB - The severe combined immunodeficient (SCID) mouse model, engrafted with human peripheral blood mononuclear cells (hu-PBMC) has proven to be useful in studying the human immune response. A major limitation of the hu-PBMC-SCID model has been the failure to consistently demonstrate a primary human immune response. Previously we developed a hu-PBMC-SCID mouse model in which we addressed both issues of adequate human lymphocyte engraftment and impaired differentiation. We demonstrated that a primary human immune response to the T-independent (TI-2) meningococcal group C capsular polysaccharide (MCPS) can be obtained in hu-PBMC SCID mice by the administration of human cytokines. In this study we compared the V(H) sequence of the MCPS response generated by B cells derived from a volunteer in the SCID mouse model to those generated by the donors' B cells in vivo. Human peripheral blood mononuclear cells were recovered from MCPS immunized hu-PBMC SCID mice and immunized donor. B cells with specificity for MCPS were isolated from these cell preparations using an anti-idiotypic monoclonal antibody which mimics MCPS. Immunoglobulin mRNA was isolated from single cells, amplified by the polymerase chain reaction, cloned and sequenced. We analysed a total of 15 V(H) regions from B cells obtained from SCID mice and a total of 13 V(H) regions from B cells obtained from the immunized donor. The response differed between SCID and in vivo cells, when studied at the genetric level. V, D and J gene usage was markedly different, however canonical structures of the hypervariable loops were conserved. The complementary determining region 3 (CDR3) varied, such that SCID derived sequences encoded longer CDR3 s than those of the donor. However all CDR3 s were rich in hydrophobic amino acids, most notably tyrosine and tryptophan, a characteristic common to many carbohydrate binding antibodies. PMID- 10378684 TI - The murine p75 TNF receptor promoter region: DNA sequence and characterization of a cis-acting silencer. AB - The promoter region of the murine p75 TNF receptor (TNF-R) was isolated from a mouse genomic DNA cosmid library. The promoter region is devoid of TATA box and has the characteristics of a house keeping gene since it contains multiple SP1 binding sites. Its mRNA has different initiation sites in different lymphoid cell lines. This promoter confers transcriptional activity to heterologous reporter genes. Deletion analysis showed that there is a silencer element located upstream from position -841. This inhibitory sequence is active both in fibroblasts and T cell lines transiently or permanently transfected. A fragment from -929 to -841 is capable of transferring this 'silencer' activity to the early SV40 promoter. This activity could be blocked in trans- when a plasmid containing the same sequence was co-transfected with the reporter plasmid indicating that a protein binds to this region of the promoter. PMID- 10378685 TI - Murine B-cell and T-cell epitopes of the allergen Hev b 5 from natural rubber latex. AB - Hev b 5, a proline-rich acidic protein with a predominantly random secondary structure, is a major allergen in natural rubber latex and a candidate for specific immunotherapy of latex allergy. As a first step in the identification of candidate peptides for immunotherapy, we have begun to identify the B-cell and T cell epitopes of Hev b 5 in BALB/c mice. The mice were immunized with a Hev b 5 fusion protein. The B-cell epitopes were determined by the SPOTS method using overlapping octamers or by ELISA inhibition using a series of overlapping 20 mers. The T-cell epitopes were determined by the proliferation and cytokine release of splenocytes cultured in the presence of the 20-mers. Potential antibody binding regions included residues in regions 1-38, 55-74, 109-128 and 132-151. Examination of the binding sequences for common motifs suggested enhanced antibody binding to the KXEE or KEXE sequences, where X is empty, threonine or alanine. Splenocyte stimulation and cytokine release suggest T-cell epitopes with the regions 1-20, 37-56, 73-101 and 109-146. Since they may contain major T-cell epitopes but do not exhibit significant antibody binding, peptide regions 38-48 and 75-101 are candidates for specific immunotherapy to Hev b 5 in the BALB/c mouse model. PMID- 10378686 TI - Analysis of the C5a anaphylatoxin core domain using a C5a phage library selected on differentiated U937 cells. AB - The human anaphylatoxin C5a is a 74-amino acid comprising polypeptide with a plethora of biological functions. Site directed mutagenesis studies suggest that several residues within the core and the C-terminus mediate the interaction with the C5a receptor. However, the contribution of particular core residues to receptor binding remained to be clarified. By means of the phage display technique, the loop between positions 35-40 was randomly mutated and the resulting C5a[35-40] fusion phage library affinity selected on C5a receptor expressing U937 cells. After five rounds of affinity enrichment, residues Arg37 and Arg40 were preferably selected. Enrichment was as high as 100% for Arg37 and 79% for Arg40. No significant enrichment of consensus residues could be obtained for positions 35, 36, 38 and 39. The core mutant C5a[A35E36R37A38S39R40], in which only Arg37/40 and Ala38 are of the native C5a sequence, was as potent as native C5a in both receptor binding and enzyme release examined on U937 cells. In contrast, replacement of Arg40 as in the mutant C5a[Q35E36R37I38L39N40] resulted in a 10-fold decrease in both binding and functional activities. Thus, selected out of a multiplicity of possibilities by the natural binding partner, Arg37 as well as Arg40 appear to be anchor residues in binding to the C5a receptor. PMID- 10378687 TI - Myc and Myb: are the veils beginning to lift? PMID- 10378688 TI - The Myc oncoprotein: a critical evaluation of transactivation and target gene regulation. AB - Mutations which disrupt the regulation or expression level of the c-myc gene are among the most common found in human and animal cancers (reviewed in ref. Cole, 1986; Henriksson and Luscher, 1996; Marcu et al., 1992). Ectopic expression studies define numerous biological activities of the c-myc gene, including transformation, immortalization, blockage of cell differentiation and induction of apoptosis (Askew et al., 1991; Cole, 1986; Evan and Littlewood, 1993; Freytag et al., 1990; Henriksson and Luscher, 1996; Marcu et al., 1992). Furthermore, c myc is required for efficient progression through the cell cycle (Goruppi et al., 1994; Prochownik et al., 1988; Yokoyama and Imamoto, 1987), although recent studies indicate that it is not absolutely essential (Mateyak et al., 1997). This fascinating array of biological activities makes the c-myc gene one of the most intriguing oncogenes and presents the challenging question of how a single gene can manifest so many different effects. The c-Myc protein exhibits sequence specific DNA binding when dimerized with its partner Max, and DNA binding is mediated through the basic region, which recognizes the core sequence CACGTG (Berberich et al., 1992; Blackwell et al., 1993; Blackwood and Eisenman, 1991; Prendergast and Ziff, 1991; Prendergast et al., 1991), but exhibits somewhat higher affinity for the more extended sequence ACCACGTGGT (Berberich et al., 1992; Blackwell et al., 1993; Halazonetis and Kandil, 1991). There are three closely related Myc family proteins (c-Myc, N-Myc and L-Myc), each with documented oncogenic potential (Birrer et al., 1988; Schwab et al., 1985; Yancopoulos et al., 1985) and similar DNA binding properties (Mukherjee et al., 1992). For simplicity, we will use the term Myc to refer to all three proteins, but delineate any distinct activities where they apply. The goal of this review is to discuss Myc as a transcriptional activator and critically evaluate the evidence for the transactivation of specific target genes as direct downstream effectors. Since excellent comprehensive reviews on Myc have been published recently (Facchini and Penn, 1998; Henriksson and Luscher, 1996), we will focus on the latest observations that offer mechanistic insight into transactivation and oncogenic transformation. PMID- 10378689 TI - Myc-mediated transformation: the repression connection. AB - Myc is an important regulator of many cellular processes, including growth promotion, differentiation, and apoptosis. The mechanisms underlying Myc biological activity, however, remain elusive. For many years, research in the field has focused on the idea of Myc as a transactivator of gene expression. More recently, alternative mechanisms of Myc function have been proposed, including gene repression. In this review we present several lines of evidence to support a connection between Myc-mediated transformation and transcriptional repression. PMID- 10378690 TI - Mysterious liaisons: the relationship between c-Myc and the cell cycle. AB - A large body of physiological evidence shows that either upregulation or downregulation of intracellular c-Myc activity has profound consequences on cell cycle progression. Recent work suggests that c-Myc may stimulate the activity of cyclin E/cyclin-dependent kinase 2 (Cdk2) complexes and antagonize the action of the Cdk inhibitor p27KIP1. Cyclin D/Cdk4/6 complexes have also been implicated as targets of c-Myc activity. However, in spite of considerable effort, the mechanisms by which c-Myc interacts with the intrinsic cyclin/Cdk cell cycle machinery remain undefined. PMID- 10378691 TI - New Myc-interacting proteins: a second Myc network emerges. AB - Despite its intensive investigation for almost two decades, c-Myc remains a fascinating and enigmatic subject. A large and compelling body of evidence indicates that c-Myc is a transcription factor with central roles in the regulation of cell proliferation, differentiation, and apoptosis, but its exact function has remained elusive. In this review we survey recent advances in the identification and analysis of c-Myc-binding proteins, which suggest insights into the transcriptional roles of c-Myc but which also extend the existing functional paradigms. The C-terminal domain (CTD) of c-Myc mediates interaction with Max and physiological recognition of DNA target sequences, events needed for all biological actions. Recently described interactions between the CTD and other cellular proteins, including YY-1, AP-2, BRCA-1, TFII-I, and Miz-1, suggest levels of regulatory complexity beyond Max in controlling DNA recognition by c Myc. The N-terminal domain (NTD), which includes the evolutionarily conserved and functionally crucial Myc Box sequences (MB1 and MB2), contains the transcription activation domain (TAD) of c-Myc as well as regions required for transcriptional repression, cell cycle regulation, transformation, and apoptosis. In addition to interaction with the retinoblastoma family protein p107, the NTD has been shown to interact with alpha-tubulin and the novel adaptor proteins Binl, MM-1, Pam, TRRAP, and AMY-1. The structure of these proteins and their effects on c-Myc actions suggest links to the transcriptional regulatory machinery as well as to cell cycle regulation, chromatin modeling, and apoptosis. Investigations of this emerging NTD-based network may reveal how c-Myc is regulated and how it affects cell fate, as well as providing tools to distinguish the physiological roles of various Myc target genes. PMID- 10378692 TI - The basic region/helix-loop-helix/leucine zipper domain of Myc proto oncoproteins: function and regulation. AB - A large body of evidence has been accumulated that demonstrates dominant effects of Myc proto-oncoproteins on different aspects of cellular growth. Myc is one of the few proteins that is sufficient to drive resting cells into the cell cycle and promote DNA synthesis. In line with this finding is that the constitutive expression of Myc in cells blocks their differentiation. These growth stimulating properties are most likely responsible for Myc's ability to initiate and promote tumor formation. Interestingly Myc can also sensitize cells to apoptosis, suggesting that this protein is part of a life-and-death switch. Molecularly Myc functions as a transcriptional regulator that needs to heterodimerize with Max to exert the biological activities described above and to regulate gene transcription. Myc and Max are just two members of a growing family of proteins referred to as the Myc/Max/Mad network. A hallmark of these proteins is that they possess a C-terminal basic region/helix-loop-helix/leucine zipper domain (bHLHZip). The bHLHZip domain specifies dimerization within the network and determines sequence specific DNA binding. Importantly this domain together with the N-terminal transactivation domain is essential for Myc biology. Here we have summarized the structural, functional, and regulatory aspects of the bHLHZip domain of Myc proteins. PMID- 10378693 TI - Mechanisms of apoptosis by c-Myc. AB - Much recent research on c-Myc has focused on how it drives apoptosis. c-Myc is widely known as a crucial regulator of cell proliferation in normal and neoplastic cells, but until relatively recently its apoptotic properties, which appear to be intrinsic, were not fully appreciated. Its death-dealing aspects have gained wide attention in part because of their potential therapeutic utility in advanced malignancy, where c-Myc is frequently deregulated and where novel modalities are badly needed. Although its exact function remains obscure, c-Myc is a transcription factor and advances have been made in characterizing target genes which may mediate its apoptotic properties. Candidate regulators and effectors are also emerging. Among recent findings are connections to the CD95/Fas and TNF pathways and roles for the tumor suppressor p19ARF and the c-Myc interacting adaptor protein Binl in mediating cell death. In this review I summarize the data establishing a role for c-Myc in apoptosis in diverse settings and present a modified dual signal model for c-Myc function. It is proposed that c-Myc induces apoptosis through separate 'death priming' and 'death triggering' mechanisms in which 'death priming' and mitogenic signals are coordinated. Investigation of the mechanisms that underlie the triggering steps may offer new therapeutic opportunities. PMID- 10378694 TI - The role of c-myc in cellular growth control. AB - Cell division is coupled to cell growth. Since some c-myc target genes are regulators of cell growth while others function in cell division pathways, c-myc is apparently poised at the interface of these processes. Cell culture systems have shown specific myc-associated growth phenotypes. Increased cell growth precedes DNA synthesis after myc activation in cells expressing myc-estrogen receptor fuson constructs and cells lacking c-myc exhibit a marked loss of protein synthesis. A number of candidate c-myc target genes regulate processes required for cell growth including rRNA transcription and processing, ribosomal protein transcription and translation, and translation initiation. These interactions all have the potential to account for the growth phenotypes in c-myc mutant cells. The ability of translation initiation factors, including eIF4E, to transform cells makes them particularly interesting targets of c-myc. Further evaluation of these target genes will provide important insights into growth control and c-myc's functions in cellular proliferation. PMID- 10378695 TI - The v-myc oncogene. AB - v-myc is the viral homolog of c-myc transduced by several acute transforming retroviruses, many of which encode this gene as a Gag-Myc fusion protein. The v myc oncogene can transform several lineages of mammalian and avian cells either alone or in cooperation with other oncogenes. While the Gag portion of the Gag Myc fusion protein and the nuclear localization signal each appear to be dispensable for transformation, the N- and C-termini of the Myc sequence have been found to be essential for transformation. All v-myc genes contain point mutations which seem to confer a greater potency to v-myc in the process of transformation, proliferation, and apoptosis. In v-myc-transformed myelomonocytic cells, secondary events occur, such as the expression of colony stimulating factor-1 (CSF-1) which play a critical role in immortalization and subsequent tumor progression. Inhibition of the autocrine loop of CSF-1 was found to induce apoptosis in the immortalized cells. While overexpression of v-Myc blocks terminal differentiation of hematopoietic cells, this is not sufficient to block the differentiation of certain neural and skeletal muscle cells. Recent developments on the effects of v-myc on cell growth, transformation, differentiation and apoptosis are discussed in this review. PMID- 10378696 TI - MYC oncogenes and human neoplastic disease. AB - c-myc, N-myc and L-myc are the three members of the myc oncoprotein family whose role in the pathogenesis of many human neoplastic diseases has received wide empirical support. In this review, we first summarize data, derived mainly from non-clinical studies, indicating that these oncoproteins actually serve quite different roles in vivo. This concept necessarily lies at the heart of the basis for the observation that the deregulated expression of each MYC gene is reproducibly associated with only certain naturally occurring malignancies in humans and that these genes are not interchangeable with respect to their aberrant functional consequences. We also review evidence implicating each of the above MYC genes in specific neoplastic diseases and have attempted to identify unresolved questions which deserve further basic or clinical investigation. We have made every attempt to review those diseases for which significant and confirmatory evidence, based on studies with primary tumor material, exists to implicate MYC members in their causation and/or progression. PMID- 10378697 TI - The myb gene family in cell growth, differentiation and apoptosis. AB - The myb gene family consists of three members, named A, B and c-myb which encode nuclear proteins that function as transcriptional transactivators. Proteins encoded by these three genes exhibit a tripartate structure with an N-terminal DNA-binding domain, a central transactivation domain and a C-terminal regulatory domain. These proteins exhibit highest homology in their DNA binding domains and appear to bind DNA with overlapping sequence specificities. Transactivation by myb gene family varies considerably depending on cell type and promoter context suggesting a dependence on interaction with other cell type specific co-factors. While the C-terminal domains of A-Myb and c-Myb proteins exert a negative regulatory effect on their transcriptional transactivation function, the C terminal domain of B-Myb appears to function as a positive regulator of this activity. One or more of these proteins interact with other transcription factors such as Ets-2, CEBP and NF-M. In addition, expression of these genes is cell cycle-regulated and inhibition of their expression with antisense oligonucleotides has been found to affect cell cycle-progression, cell division and/or differentiation. Members of the myb gene family exhibit different temporal and spatial expression patterns suggesting a distinctive function for each of these genes. Gene knockout experiments show that these genes play an essential role in development. Loss of c-myb function results in embryonic lethality due to failure of fetal hepatic hematopoiesis. A-myb null mutant mice, on the other hand are viable but exhibit growth abnormalities, and defects in spermatogenesis and female breast development. While the role of c-myb in oncogenesis is well established, future experiments are likely to provide further clues regarding the role of A-myb and B-myb in tumorigenesis. PMID- 10378698 TI - Reassessing the role of C-MYB in tumorigenesis. AB - Hematopoietic tumors in both humans and mice frequently up-regulate expression of the c-myb gene, but it is unclear whether this is a cause or a consequence of the leukemic state. Recent results placing super-activation of the c-Myb protein at the bottom of a kinase-activated signal transduction pathway indicate that it may be a downstream effector of transformation induced by other oncogenes. The relationship between c-Myb and the serine-threonine kinase pim-1, its immediate activator, is discussed, together with the possibility that c-Myb, like pim-1, may be able to synergize with c-Myc to induce tumors. PMID- 10378699 TI - Myb binding proteins: regulators and cohorts in transformation. AB - The c-Myb and v-Myb proteins are transcription factors that regulate cell proliferation and differentiation. Both Myb proteins have been shown to interact with a number of cellular proteins, some of which are transcription factors that cooperate to activate specific promoters, while others regulate the transcriptional activity of Myb in specific contexts. By comparing and analysing the types of proteins that bind Myb, and the conserved domains of Myb that interact with other proteins, conclusions can be drawn regarding the role of specific partner proteins in the regulation of gene expression, cell proliferation and disease. PMID- 10378700 TI - Transformation by v-Myb. AB - The v-myb oncogene of the avian myeloblastosis virus (AMV) is unique among known oncogenes in that it causes only acute leukemia in animals and transforms only hematopoietic cells in culture. AMV was discovered in the 1930s as a virus that caused a disease in chickens that is similar to acute myelogenous leukemia in humans (Hall et al., 1941). This avian retrovirus played an important role in the history of cancer research for two reasons. First, AMV was used to demonstrate that all oncogenic viruses did not contain a single cancer-causing principle. In particular, although both Rous sarcoma virus (RSV) and AMV could replicate in cultures of either embryonic fibroblasts or hematopoietic cells, RSV could transform only fibroblasts whereas AMV could transform only hematopoietic cells (Baluda, 1963; Durban and Boettiger, 1981a). Second, chickens infected with AMV develop remarkably high white counts and therefore their peripheral blood contains remarkably large quantities of viral particles (Beard, 1963). For this reason AMV was often used as a prototypic retrovirus in order to study viral assembly and later to produce large amounts of reverse transcriptase for both research and commercial purposes. Following the discovery of the v-src oncogene of RSV and the demonstration that it arose from the normal c-src proto-oncogene, a number of acute leukemia viruses were analysed by similar techniques and found to also contain viral oncogenes of cellular origin (Roussel et al., 1979). In the case of AMV, it was shown that almost the entire retroviral env gene had been replaced by a sequence of cellular origin (initially called mab or amv, but later renamed v-myb) (Duesberg et al., 1980; Souza et al., 1980). Remarkably, sequences contained in this myb oncogene were shared between AMV and the avian E26 leukemia virus, but were not contained in any other acutely transforming retroviruses. In addition, the E26 virus contained a second sequence of cellular origin (ets) that was unique. The E26 leukemia virus was first described in the 1960s and causes an acute erythroblastosis in chickens, more reminiscent of the disease caused by avian erythroblastosis virus (AEV) than by AMV (Ivanov et al., 1962). PMID- 10378702 TI - Interleukin 10 and arthritis. PMID- 10378701 TI - Myb targeted therapeutics for the treatment of human malignancies. AB - For the past several years, we have been engaged in developing a therapeutically effective strategy for disrupting gene function with reverse complementary, or so called 'antisense', oligodeoxynucleotides (ODN). This pursuit has focused on finding appropriate diseases in which to apply this approach, and suitable gene targets. Of the genes that we have targeted for disruption using the antisense ODN strategy (Clevenger et al., 1995; Gewirtz and Calabretta, 1988; Ratajczak et al., 1992c; Small et al., 1994) one that has been of particular interest, and one where therapeutically motivated disruptions are now in clinical trial, is the myb gene (reviewed in Lyon et al., 1994). These trials involve treatment of human leukemias. These diseases are a logical choice for developing oncogene targeted therapies because of easy access to tissues, and the abundance of knowledge about the cell and molecular biology of these diseases. Nevertheless, as will be touched on below, other malignancies have also been examined as models for Myb targeted therapy with some surprisingly encouraging results. Finally, while we have focused our efforts on the ODN strategy, I will allude briefly to other strategies for disrupting Myb function with therapeutic intent. PMID- 10378703 TI - Prolactin, the immune response and lupus. PMID- 10378704 TI - Is there a defect in cortisol production in rheumatoid arthritis? PMID- 10378705 TI - A 12-month randomized controlled trial of patient education on radiographic changes and quality of life in early rheumatoid arthritis. AB - OBJECTIVE: In rheumatoid arthritis, education programmes successfully impart knowledge but, notwithstanding issues of empowerment, this knowledge has to be translated into behavioural change to have a chance of improving disease outcome. Arguably, behavioural change must also occur early if outcomes are to be improved. For these reasons, we planned a study of patient education in early disease, with radiological damage and quality of life as the main outcome variables. METHODS: We performed a randomized controlled trial in people with rheumatoid arthritis of < 5 yr duration. The main intervention was a 4 week education programme, each weekly session lasting 2 h. Assessments were made at entry, at 4 weeks and at 12 months. The main outcome variables were the modified Larsen radiological score for the hands and the SF-36 quality of life questionnaire. Secondary outcome variables were the Health Assessment Questionnaire (HAQ), Ritchie Articular Index (RAI), Patient Knowledge Questionnaire (PKQ), Compliance Questionnaire (CQ), plasma viscosity (PV), pharmaceutical changes and consulting behaviour. RESULTS: The patient numbers were 34 (10 male, 24 female) for the control group and 43 (16 male, 27 female) for the education group. The groups were matched for age (56.5 yr for control, 55 yr for education), disease duration (3.5 yr vs 3.0 yr) and duration of second line drug therapy (14 months vs 12 months). We found no significant difference between the groups for Larsen scores at 12 months, although scores for the education group were lower (39.5 vs 43.0, P = 0.13). The 'social functioning' and 'general health perception' subscales of the SF-36 showed a significant improvement in the education group, but no significant differences between groups were seen. No significant differences were found for the HAQ, RAI, PV and CQ, but the education group had more disease-specific knowledge than the control group at 12 months (PKQ scores: 17 vs 21, P = 0.0002). No differences were found for out patient visits and in-patient admissions, but the education group had slightly more changes in second-line drugs during the study (0.43 changes/person in the control group, 0.51 changes/person in the education group). CONCLUSIONS: We found no significant difference between the groups in our primary outcome measures, but a trend in favour of the education group was found in radiological progression. Further studies of this kind, using larger patient numbers, are required since the difference may result from improved self-care, better compliance with joint protection strategies and, possibly, improved drug compliance. PMID- 10378706 TI - Probucol improves symptoms and reduces lipoprotein oxidation susceptibility in patients with Raynaud's phenomenon. AB - OBJECTIVE: Reactive oxygen species have been implicated in the pathogenesis of inflammatory and vascular disease. We have undertaken a controlled trial to evaluate probucol, a synthetic antioxidant, as a potential therapy for Raynaud's phenomenon. METHODS: The study cohort included patients with systemic sclerosis (SSc; n = 20), primary Raynaud's phenomenon (n = 15) or 'autoimmune Raynaud's' (n = 5). Patients were allocated to receive either probucol (500 mg daily) or nifedipine (20 mg daily) for 12 weeks. Clinical and biochemical variables at baseline were compared with those at completion of treatment. Evaluation included assessment of Raynaud's attack frequency and severity by visual analogue scale, measurement of low-density lipoprotein (LDL) oxidation lag time, and plasma concentrations of cholesterol, triglyceride, vitamin E and vitamin C. RESULTS: There was a significant reduction of both the frequency and severity of Raynaud's attacks in the patients who received probucol, but not in the control group. LDL oxidation lag time, reflecting in vitro susceptibility to oxidation, was also increased by probucol therapy and serum cholesterol levels were significantly reduced. Similar changes were observed in both SSc- and non-SSc-associated Raynaud's cases. CONCLUSION: These data suggest that probucol may be useful for the symptomatic treatment of Raynaud's phenomenon and also reduces LDL oxidation susceptibility. Since oxidized lipoproteins may mediate vascular damage in SSc, the use of probucol could have additional disease-modifying benefits. Based upon the results of this pilot study, further evaluation of this novel form of therapy is warranted. PMID- 10378708 TI - Risk taking in patients with rheumatoid arthritis: are the risks of haemopoietic stem cell transplantation acceptable? AB - OBJECTIVES: Autologous haemopoietic stem cell transplantation (HSCT), which carries defined risks of early treatment-related mortality (TRM), has recently been proposed as an experimental therapy for severe rheumatoid arthritis (RA). The aim of this study was to establish whether the risks of this approach are acceptable to patients with RA and whether risk taking related to disease associated or personal/social parameters. METHODS: A standard gamble questionnaire was used to determine the acceptable risk of mortality for a potentially curative procedure in patients with RA aged <70 yr. Additional data collected included age, sex, duration of RA, number of second-line agents, domestic and workforce information, and self-assessed disability. RESULTS: The 53 patients (age range 24-69 yr, 39 female, 14 male, disease duration 2-43 yr) interviewed were prepared to accept a broad range of treatment-related mortality in order to be returned to normality off all drugs (median 5%, range 0-50%). Risk taking was significantly related to degree of disability measured by the disability section of the Health Assessment Questionnaire (HAQ; P = 0.001) and negatively related to age (P = 0.04), although only HAQ score maintained significance on multivariate analysis. Using linear regression, we were able to determine that current TRM of autologous HSCT in Australia (3.3%) would be acceptable to patients with HAQ scores of >0.44 (84% of our sample), but allogeneic HSCT (with a TRM of 13.1%) would be acceptable only to severely disabled patients with HAQ scores of >2.45 (4% of our sample), assuming the procedure to be curative. CONCLUSION: Along with previous studies, these results suggest that, if long-term efficacy can be proven, then the risks of autografting may be acceptable to most patients with RA, particularly those with significant disability. PMID- 10378707 TI - Chromosomal analysis of peripheral lymphocytes of patients before and after radiation synovectomy with samarium-153 particulate hydroxyapatite. AB - OBJECTIVE: Radiation synovectomy may be indicated for the treatment of chronic synovitis. A number of factors may affect its current use, including availability, limited evidence for its efficacy compared to intra-articular glucocorticoid, and concerns regarding the potential long-term effects of radiation exposure, particularly in younger patients. Specific chromosome-type abnormalities in peripheral lymphocytes can be useful indicators of whole-body radiation exposure. The frequency of these aberrations has been shown to increase in patients who have had radiation synovectomy using yttrium-90 by up to five times compared to baseline levels. Samarium-153 particulate hydroxyapatite (Sm 153 PHYP) is a new radiopharmaceutical currently on trial which appears to have less extra-articular leakage than yttrium-90 compounds. The aim of this study was to identify any increase in specific chromosome-type abnormalities, using published criteria, in patients following Sm-153 PHYP synovectomy of the knee. The 10 patients (five men, five women) in whom the analyses were performed had a mean age of 47 yr (range 28-70 yr). RESULTS: There was no increase in scored chromosome-type abnormalities after Sm-153 PHYP synovectomy. CONCLUSION: This study further supports the relative safety of Sm-153 PHYP compared to other radiopharmaceuticals. PMID- 10378709 TI - CD5 and CD23 expression on B cells in peripheral blood and synovial fluid of rheumatoid arthritis patients: relationship with interleukin-4, soluble CD23 and tumour necrosis factor alpha levels. AB - METHODS: We have studied in peripheral blood (PB) and synovial fluid (SF) of 31 patients diagnosed with rheumatoid arthritis (RA), the expression of CD5 and CD23 antigens on B cells, and the levels of soluble CD23 (sCD23), interleukin-4 (IL-4) and tumour necrosis factor alpha (TNF-alpha). We have also correlated the results with the disease activity index. RESULTS: CD5+ B cells are expanded in SF and, moreover, show higher expression of CD23 than CD5 - B cells. Twelve patients had detectable levels of IL-4 in plasma and 10 in SF (nine patients in both samples); the absence of IL-4 was related to a higher expression of CD23 on CD5 + B cells and with higher levels of sCD23. A negative correlation was found in SF between TNF-alpha and sCD23 levels. CONCLUSION: There is no correlation between disease activity index and the different parameters studied (expression of CD5 and CD23 on B cells, sCD23, IL-4 and TNF-alpha levels) either in plasma/PB or in SF. PMID- 10378710 TI - A preliminary study of ultrasound aspiration of bone erosion in early rheumatoid arthritis. AB - OBJECTIVE: To develop a new technique to assess the primary lesion in early rheumatoid arthritis (RA). METHODS: Ten patients with early RA and radiographically or MRI confirmed erosions had a needle introduced into the base of the erosion under sonographic guidance. Material was then aspirated from this site. RESULTS: The procedure was well tolerated with no complications. Small samples of necrotic bone and tissue were obtained in five out of 10 cases. In one case, a distinctive population of pleomorphic CD34 + cells with characteristics of bone marrow progenitors was isolated. Tissue invading bone with a characteristic appearance of pannus was not seen. CONCLUSION: A new method of sampling the earliest lesion in RA is described. The findings raise questions about the nature of bone damage in early RA. PMID- 10378711 TI - Clinical, radiographic and HLA associations as markers for different patterns of psoriatic arthritis. AB - OBJECTIVE: The aim of this study was to examine whether the five clinical forms of psoriatic arthritis (PsA) identified by Moll and Wright (Semin Arthritis Rheum 1973;3:55-78) could be clearly distinguished, especially as the disease evolved over time, to analyse whether radiographic features or HLA associations could define subsets with greater precision and to identify predictors of disease outcome. METHODS: Seventy-three patients (37 males and 36 females) were followed for a median time of 8 yr (range 1-16 yr). A standard clinical protocol was used to assess patients at each visit and two clinical scores. based on the joint areas involved, were defined to evaluate the mode of onset and the evolution of arthritis. X-ray films of the hands, feet and sacroiliac joints were taken and the patients were divided into two categories according to the presence or absence of erosions and an X-ray erosion score was also used. Three classification methods were used to define the different clinical subsets. HLA-A, B and DR antigens were tested by standard microlymphocytotoxicity assays. A multiple linear regression model was used in the statistical analysis. RESULTS: The five classical clinical subsets defined by Moll and Wright did not remain since distinct peripheral arthritis patterns tended to evolve over time. Only two discrete groups were identified, axial disease (AD) (sacroilitis with or without peripheral arthritis) in 29% of cases and peripheral disease (PD) without sacroilitis in 71%. AD was positively associated with the duration of arthritis (P < 0.04), presence of mutilation (P < 0.02) and the joint area score over disease evolution (JASE) (P < 0.02). There were erosions in 71% of the patients. Erosions correlated with the presence of mutilation (P < 0.007) and with the JASE (P < 0.0005). HLA-B27 was found in 43% of patients with AD, but only in 11% of PD patients (P < 0.01). No other clear HLA correlations were found. CONCLUSIONS: Despite the relatively small number of patients, this longitudinal study suggests that only two clinical subsets can be clearly defined in PsA, AD and PD; these are primarily determined on clinical grounds although HLA-B27 is strongly associated with AD. The evolution of PD pattern with time means that narrower peripheral arthritis subsets are of little clinical use. PMID- 10378712 TI - Type IX collagen immunoreactive peptides in synovial fluids from arthritis patients. AB - OBJECTIVES: To determine whether type IX collagen-related peptides can be detected in the synovial fluids of arthritis patients and to assess their potential as molecular markers of arthritis. PATIENTS/METHODS: Synovial fluids from a set of carefully diagnosed arthritis patients and from healthy volunteers were used. Hydroxyproline assays were carried out to determine the content and concentration of collagen. Collagen cross-link determinations were conducted by reversed-phase HPLC. SDS PAGE and immunoblotting were used to identify the collagenous components, and N-terminal sequencing was performed to confirm these identities. RESULTS: All the synovial fluids were found to contain measurable amounts of collagen at similar concentrations. This appeared to be mainly high molecular-weight material consisting of type I and type IX collagens, but not type II collagen. However, other smaller molecular weight type IX immunoreactive peptides were detected which were more apparent in the synovial fluids from arthritis patients. These peptides were also found to contain non-collagenous material. Collagen cross-links were also present in the arthritis synovial fluids. CONCLUSION: Collagenous material can be detected in all synovial fluids and the presence of pyridinoline cross-links indicates that at least some of this is derived from a mature collagen matrix. Type IX immunoreactive peptides were identified, but were found to contain significant amounts of non-collagenous material, and their presence, even at lower levels, in synovial fluids from normal subjects limits their potential for use as molecular markers of disease. Nevertheless, this is the first report of type IX collagen-related fragments in synovial fluids. PMID- 10378713 TI - A randomized trial of acupuncture as an adjunctive therapy in osteoarthritis of the knee. AB - OBJECTIVE: The purpose of this study was to investigate the efficacy of acupuncture as an adjunctive therapy to standard care for the relief of pain and dysfunction in elderly patients with osteoarthritis (OA) of the knee. METHODS: Seventy-three patients with symptomatic OA of the knee were randomly assigned to treatment (acupuncture) or standard care (control). Analysis was performed on last score carried forward to account for patients who dropped out before completion. Patients self-scored Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) and Lequesne indices at baseline and at 4, 8 and 12 weeks. Patients in the control group were offered acupuncture treatment after 12 weeks. The data for these patients are pooled with those from the original acupuncture group for within-group analysis. RESULTS: Patients randomized to acupuncture improved on both WOMAC and Lequesne indices compared to those who received standard treatment alone. Significant differences on total WOMAC Scale were seen at 4 and 8 weeks. There appears to be a slight decline in effect at 4 weeks after cessation of treatment (12 weeks after first treatment). No adverse effects of acupuncture were reported. CONCLUSION: These data suggest that acupuncture is an effective and safe adjunctive therapy to conventional care for patients with OA of the knee. PMID- 10378714 TI - Determinants of WOMAC function, pain and stiffness scores: evidence for the role of low back pain, symptom counts, fatigue and depression in osteoarthritis, rheumatoid arthritis and fibromyalgia. AB - OBJECTIVES: The Western Ontario MacMaster (WOMAC) is a validated instrument designed specifically for the assessment of lower extremity pain and function in osteoarthritis (OA) of the knee or hip. In the clinic, however, we have noted that OA patients frequently have other musculoskeletal and non-musculoskeletal problems that might contribute to the total level of pain and functional abnormality that is measured by the WOMAC. In this report, we investigated back pain and non-articular factors that might explain WOMAC scores in patients with OA, rheumatoid arthritis (RA) and fibromyalgia (FM) in order to understand the specificity of this instrument. METHODS: RA, OA and FM patients participating in long-term outcomes studies completed the WOMAC and were assessed for low back pain, fatigue, depression and rheumatic disease symptoms by mailed questionnaires. RESULTS: Regardless of diagnosis, WOMAC functional and pain scores were very much higher (abnormal) among those complaining of back pain. On average, WOMAC scores for back pain (+) patients exceeded those of back pain (-) patients by approximately 65%,, and 52% of OA patients reported back pain. In regression analyses, study symptom variables explained 42, 44 and 38% of the variance in WOMAC function, pain and stiffness scores, respectively. In the subset of OA patients, radiographic scores added little to the explained variance. The strongest predictor of WOMAC abnormality in bivariate and multivariate analyses was the fatigue score, with correlations of 0.58, 0.60 and 0.53 with WOMAC function, pain and stiffness, respectively. The WOMAC performed well in RA and FM, and correlated strongly with the Health Assessment Questionnaire (HAQ) disability scale and a visual analogue scale (VAS) pain scale. CONCLUSION: The WOMAC captures more than just knee or hip pain and dysfunction, and is clearly influenced by the presence of fatigue, symptom counts, depression and low back pain. WOMAC scores also appear to reflect psychological and constitutional status. These observations suggest the need for care in interpreting WOMAC scores as just a measure of function, pain or stiffness, and indicate the considerable importance of psychological factors in rheumatic disease and rheumatic disease assessments. PMID- 10378716 TI - An uncommon association: Sjogren's syndrome and autoimmune myelofibrosis. PMID- 10378715 TI - The effects of interferon beta treatment on arthritis. AB - OBJECTIVE: To determine whether interferon beta (IFN-beta) therapy might have a beneficial effect on arthritis, we evaluated the effect of IFN-beta on collagen type II-induced arthritis (CIA) in rhesus monkeys and conducted a pilot study in patients with rheumatoid arthritis (RA). METHODS: Four rhesus monkeys with CIA were treated with 10 x 10(6) U (MIU)/kg mammalian cell-derived recombinant IFN beta (Rebif; Ares-Serono) s.c. daily for 1 week. Subsequently, 12 patients with active RA were treated for 12 weeks with purified natural fibroblast IFN-beta (Frone, Ares-Serono) s.c. 3 times weekly at the following dosages: 6 MIU (n = 4), 12 MIU (n = 4) and 18 MIU (n = 4). RESULTS: Rapid clinical improvement during IFN beta therapy was observed in three of the four rhesus monkeys with CIA. There was also a marked decrease in serum C-reactive protein (CRP) levels with a subsequent increase after discontinuation of the treatment in all monkeys. The 10 RA patients who completed the study exhibited on average gradual improvement of tender and swollen joint counts, patient's assessment of pain, and patient's and doctor's global assessment (all P < 0.05). The health assessment questionnaire and serum CRP levels also tended to decrease, but this was not statistically significant; 40% of the patients fulfilled the ACR criteria for 20%, improvement, whereas none fulfilled the ACR criteria for 50% improvement 12 weeks after initiation of treatment. There was no clear dose response relationship. CONCLUSION: The data suggest that IFN-beta treatment has a beneficial effect on arthritis. PMID- 10378717 TI - Bone mineral density in scleroderma. PMID- 10378718 TI - Does parenteral oestrogen therapy flare up disease activity in patients with systemic lupus erythematosus complicated by haemorrhagic cystitis? PMID- 10378719 TI - Seronegative oligoarthritis in the course of refractory anaemia with excess blasts. PMID- 10378720 TI - Hypertrophic osteoarthropathy associated with bacterial endocarditis. PMID- 10378721 TI - Cutaneous necrotizing vasculitis, erythema nodosum and ankylosing spondylitis. PMID- 10378723 TI - A case of systemic sclerosis that developed under dexfenfluramine use. PMID- 10378722 TI - Predominant ankle arthropathy in hereditary haemochromatosis. PMID- 10378724 TI - Enhanced frequency of autoimmune congenital heart block in female offspring. PMID- 10378725 TI - Oral methotrexate: the hazard of different tablet strengths. PMID- 10378726 TI - Automatic detection of cyclic alternating pattern (CAP) sequences in sleep: preliminary results. AB - OBJECTIVES: The analysis of cyclic alternating pattern (CAP) provides important microstructural information on arousal instability and on EEG synchrony modulation in the sleep process. This work presents a methodology for automatic classification of the micro-organization of human sleep EEG, using the CAP paradigm. METHODS: The classification system is composed of 3 parts: feature extraction, detection and classification. The feature extraction part is an EEG generation model-based maximum likelihood estimator. The detector part for the CAP phases A and B is done by a variable length template matched filter, while the classification criteria part is implemented on a state machine ruled-based decision system. RESULTS AND CONCLUSIONS: The preliminary results of the automatic classifier on a group of 4 middle-aged adults are presented. The high agreement between the detector and visual scoring is very promising in the achievement of a fully automated scoring system, although a more exhaustive evaluation program is needed. PMID- 10378727 TI - Spectral properties of EEG fast activity ictal discharges associated with infantile spasms. AB - OBJECTIVE: The aim of this study was to evaluate the characteristics of the ictal EEG event accompanying infantile spasms. METHODS: Quantitative analysis was used, based on the application of a bivariate autoregressive (AR) parametric model; autospectra, coherence, phase functions and inter-hemispheric time differences were estimated on homologous EEG channels in 18 infants presenting with either cryptogenic or symptomatic West syndrome. RESULTS: The AR analysis of the 500 ms EEG epochs preceding spasm onset revealed the presence of a short discharge of fast activity restricted to a narrow frequency band in 13 of the 18 cases included in the study. The fast discharge peaked at 17.5+/-2.1 Hz, with rather low inter-hemispheric coherence values (0.52+/-0.17) and asymmetric amplitude on homologous EEG derivations. It persisted briefly after spasm onset, reaching a higher coherence value (0.71+/-0.16). The inter-hemispheric time difference, estimated in those cases with the coherence values significantly different from zero, ranged from 9.1 to 14.3 ms (11.4+/-1.9) in the epoch preceding spasm onset. CONCLUSION: The data obtained from the analysis of the ictal EEG events, compared with clinical and interictal EEG features, indicate that an asymmetric EEG pattern (mainly consisting of a rhythmic burst of fast activity) consistently preceded both symmetric and asymmetric spasms, thus suggesting a localized cortical origin of the ictal discharge giving rise to the spasms. PMID- 10378728 TI - Common spatial subspace decomposition applied to analysis of brain responses under multiple task conditions: a simulation study. AB - A method, called common spatial subspace decomposition, is presented which can extract signal components specific to one condition from multiple magnetoencephalography/electroencephalography data sets of multiple task conditions. Signal matrices or covariance matrices are decomposed using spatial factors common to multiple conditions. The spatial factors and corresponding spatial filters are then dissociated into specific and common parts, according to the common spatial subspace which exists among the data sets. Finally, the specific signal components are extracted using the corresponding spatial filters and spatial factors. The relationship between this decomposition and spatio temporal source models is described in this paper. Computer simulations suggest that this method can facilitate the analysis of brain responses under multiple task conditions and merits further application. PMID- 10378729 TI - EEG power changes are related to regional cerebral glucose metabolism in vascular dementia. AB - OBJECTIVE: In patients with vascular dementia (VD), the relationship between the EEG power within the 4 frequency bands and the regional metabolic disturbances was investigated. METHODS: Twenty-eight patients (age 69.0+/-6.54 years) with VD according to NINDS-AIREN criteria underwent quantitative EEG recording, according to the 10-20 system, and fluodeoxyglucose F18 positron emission tomography (PET) at resting condition within 24 h. EEG power FFT-analysis was performed for delta (2-3.5 Hz), theta (4-7.5 Hz), alpha (8-13 Hz) and beta (13.5-20 Hz) frequency bands. Regional EEG power bands were related to regional glucose metabolism in anatomically defined regions corresponding to locations of the 10-20 system. RESULTS: Correlation between slow frequency band power and glucose metabolism was found. A widespread inverse relationship of delta power to metabolism was found between various regions; additionally, delta power was negatively correlated to cerebral glucose metabolism in individual regions. Frontal theta power correlated especially with thalamic CMRglc. Alpha power correlated directly with metabolism in the occipital lobe. No significant relationships were found between beta power and metabolism. CONCLUSION: We conclude that EEG power in VD is linked to glucose metabolism, indicating specific regional dependencies. PMID- 10378730 TI - EEG power spectrum differences in early and late onset forms of Alzheimer's disease. AB - OBJECTIVE: To evaluate the relationship between some EEG spectral parameters and age of onset of Alzheimer's disease (AD). METHODS: A study on the wakefulness EEG, recorded during eyes closed and open, was carried out on 150 AD patients (NINCDS-ADRDA criteria). Fifty-two normal subjects served as controls. RESULTS: A significant prevalence of an EEG spectrum characterised by lack of a dominant peak in the 6.5-12 Hz band was found in early AD (EAD). Age of onset correlated inversely with the 1-6.5 Hz relative powers and positively with 6.5-12 Hz relative powers. A similar correlation was also found when a subgroup of mild initial AD was selected. Moreover, evidence of EEG changes peculiar to early onset AD emerged when 3 subgroups (with age of onset < or =60, range 61-69 and > or =70 years) were compared. CONCLUSION: Irrespective of the severity of disease, this study provides evidence of specific changes of wakeful EEG in patients affected by early-onset AD. PMID- 10378731 TI - Alpha activity in the human REM sleep EEG: topography and effect of REM sleep deprivation. AB - The topographical distribution of alpha activity (8.125-11.125 Hz) in the REM sleep EEG, its time course within and across REM sleep episodes, and the effects of selective REM sleep deprivation were investigated in 8 young males. Power spectra of bipolar derivations along the antero-posterior axis in the left (F3C3, C3P3, P301) and right (F4C4, C4P4, P402) hemisphere were calculated. Alpha activity increased along the antero-posterior axis in both hemispheres, and was dominant in the right hemisphere. It decreased within and across REM sleep episodes. Selective REM sleep deprivation resulted in a reduction of alpha activity in the REM sleep EEG. However, the topographical distribution and the time course were not affected. It is suggested that alpha activity in the REM sleep EEG is a marker of REM sleep homeostasis. PMID- 10378732 TI - Gamma responses and ERPs in a visual classification task. AB - OBJECTIVE: We examined event-related potentials (ERPs) and gamma range EEG activity in a visual classification task to assess which variables affect these responses. METHODS: Ten subjects silently counted the occurrence of rare Kanizsa squares (targets) among Kanizsa triangles and non-Kanizsa figures (standards). By applying a time-frequency analysis to the data and selectively calculating topographical maps of certain frequencies. RESULTS: We were able to find 3 different types of gamma responses to Kanizsa figures: an early phase-locked gamma response at 40 Hz in the N100 time range, late phase-locked gamma activity (200-300 ms) at 40 Hz and a continuous phase-locked gamma response at 80 Hz due to the monitor refresh frequency. The two 40 Hz responses were significantly higher for Kanizsa figures than for non-Kanizsa figures and within the Kanizsa figures were higher for the target figure than for the non-target. CONCLUSION: The phase-locking of these two responses, previously found also as non-phase locked activity, could be synchronized due to the monitor flicker frequency. Also, our findings suggest that the gamma responses are not solely associated with the binding of stimulus features, but reflect some processes related to target processing. PMID- 10378733 TI - Functions and sources of event-related EEG alpha oscillations studied with the Wavelet Transform. AB - OBJECTIVES: By using the Wavelet Transform, a time frequency representation with nearly optimal resolution, we studied responses to stimulation in the 'alpha' range (10 Hz). METHODS: Visual evoked responses of 10 healthy subjects were studied with 3 different stimulus types (no-task VEP, non-target and target stimulus). RESULTS: Upon all the stimulus types, event-related responses in the 10 Hz ('alpha') range were distributed in the whole scalp, best defined in the occipital locations, the responses on the anterior electrodes being less pronounced and delayed. In some subjects, these event-related responses were prolonged upon target stimulation in posterior locations. CONCLUSIONS: These results point towards a distributed origin of event-related alpha oscillations with functional relation to sensory processing, and possibly to further processes. PMID- 10378734 TI - EEG and long-term outcome of term infants with neonatal hypoxic-ischemic encephalopathy. AB - OBJECTIVE: The prognostic value of a burst suppression pattern (BSP) on the electroencephalograph (EEG) in the prediction of long-term outcome for full term newborns with hypoxic-ischemic encephalopathy (HIE) is well established. The purpose of our study was to compare the patterns of burst suppression on EEG with long-term neurological outcome in term infants with HIE. METHODS: We retrospectively analyzed all records of all full-term newborn infants born at the University of Alberta Hospital between January 1, 1991 and December 31, 1992, who had clinical evidence of HIE and had at least one EEG during the first week of life. The EEGs were reviewed and blindly subclassified into a BSP, or if the pattern was not continuous or was incomplete, a modified burst suppression pattern (MBSP), based on specified electrophysiological criteria. The long-term neurological outcome was then correlated with the EEG pattern. RESULTS: Twenty three full-term infants were studied. Fifteen had a BSP on EEG and 8 had a MBSP. Six of 15 infants with a BSP died. Of the 9 survivors with a BSP, 7 are disabled and two are normal. Of the 8 infants in the MBSP group, one infant died, two are disabled and 5 are normal. In the BSP group, 6/7 disabled infants developed cerebral palsy while in the MBSP group, only one developed cerebral palsy. CONCLUSION: The results are suggestive of a better outcome for infants with neonatal HIE and MBSP on EEG compared with those with a BSP. Subclassification of the EEG changes of neonatal HIE into BSP and MBSP may give a more accurate prediction of outcome in perinatal asphyxia and assist in discussion with parents about prognosis. PMID- 10378735 TI - A ring-shaped distribution of dipoles as a source model of induced gamma-band activity. AB - As opposed to slow waves, spontaneous and stimulus-induced oscillations in the gamma-band show no polarity reversal in cortical depth, which cannot be explained by the classical equivalent current dipole model usually proposed as a model of pyramidal cell synaptic activity. Here we propose a ring-shaped distribution of dipoles as a source model for these fast oscillations. This distribution generates a field potential that does not reverse through cortical depth. Such a geometry could correspond to horizontally oriented dendritic fields. Moreover, this distribution generates a potential field, but no, or weak, magnetic field on the scalp surface, which corresponds to the observation that visually-induced gamma-band oscillations are detectable in EEG data, but not in simultaneously recorded MEG data. PMID- 10378736 TI - Towards an objectification by classification of tinnitus. AB - OBJECTIVE: The objectives of this study are to identify the presence of tinnitus and classify its different forms, in terms of changes in noise. METHODS: Late auditory evoked responses (LAERs) were recorded from Fz in response to 1000 Hz tone bursts of various intensities, in 13 tinnitus-free subjects and in 25 tinnitus sufferers (16 bilateral and 9 unilateral tinnitus sufferers). A classification of different forms of tinnitus, in terms of changes in noise, was also undertaken. N1-P2 component amplitudes and N1 and P2 latencies were measured. RESULTS: Objective identification of the affected ear in unilateral tinnitus sufferers was found feasible on the basis of N1-P2 intensity-dependence and N1-P2 amplitude. The bilateral tinnitus group was found to differ from controls by greater intensity-dependence of the N1-P2 component and shorter N1 latency. These characteristics varied with tinnitus type: a classification on the basis of intensity-dependence and latencies proved feasible. The group of patients showing improved tinnitus in noise had greater intensity-dependence and longer N1 latency than did the group showing aggravated tinnitus in noise. CONCLUSIONS: Data are discussed in light of the inhibitory role of frontal cortex on the sensory inputs and the modulatory function of central serotonergic system on the processing of auditory information. PMID- 10378737 TI - Face-selective spectral changes in the human fusiform gyrus. AB - OBJECTIVE: To characterize ventral occipitotemporal and prefrontal EEG during cognitive processing. METHODS: Depth probes were implanted for seizure localization in 16 pharmaco-resistant epileptics. Probes penetrated from middle temporal through fusiform to lingual gyrus, and from inferior frontal to anterior cingulate gyrus. Event-related potentials (ERPs) and event-related spectral power (ERSP) were calculated during delayed recognition for faces or words. RESULTS: Face stimuli evoked a broadband fusiform ERSP increase from 5 to 45 Hz at 150-210 ms after stimulus onset. This ERSP increase was immediately followed by an ERSP decrease in the same region from 300 to 1000 ms. Both the early increased ERSP and the late decreased ERSP, were greater for faces than words. Simultaneous with the late temporal ERSP decrease, the prefrontal depth EEG displayed a low frequency (5-12 Hz) ERSP increase to face and word stimuli. CONCLUSION: Early temporal ERSP increases occur at a time when the fusiform gyrus is thought to contribute to face processing. This increase is also reflected in spectral analysis of the ERP, but the late temporal ERSP decrease and frontal ERSP increase are not. Thus, intracranial recordings in humans demonstrate event related fluctuations in EEG spectral power with clear anatomical, temporal and cognitive specificity. PMID- 10378738 TI - Negative potential shifts and the prediction of the outcome of neurofeedback therapy in epilepsy. AB - About two-thirds of epilepsy patients who learn to control their slow cortical potential shifts (SCP) reduce their seizure rate, but the remaining third does not demonstrate clinical improvement. In the present study, this finding was replicated in a group of 27 patients with focal epilepsy. We found that patients who consistently produced larger negative SCP in all conditions during the first phase of treatment, showed no decrease in seizure frequency during the six-month follow-up, as compared with the three-month baseline phase. The large negative SCP explained about one-third of the variance of the clinical outcome. Age, medication, seizure history, or the localization of focus were found to be unrelated to clinical improvement. PMID- 10378739 TI - Hippocampal sleep spindles revisited: physiologic or epileptic activity? AB - OBJECTIVE: Few reports have described sleep spindles in intracranial electrode recordings from human hippocampus. Controversy exists regarding whether hippocampal spindles represent a physiologic or epileptic phenomenon. METHODS: We reviewed hippocampal recordings in 8 subjects to characterize events resembling sleep spindles. RESULTS: In 6 subjects, events occurred exclusively during non rapid eye movement (NREM) sleep, were similar in morphology to surface spindles, occurred simultaneously or independently of surface spindles, and did not show a consistent relationship to the epileptic region. In an additional subject, a proportion of the hippocampal activity recorded differed slightly in morphology from surface spindles, was present during both NREM and rapid eye movement (REM) sleep, occurred in the same channels as isolated interictal epileptiform discharges, attenuated just prior to seizures, and occurred postictally as repetitive discharges. This activity occurred simultaneously or independently of surface spindles, but differed from surface spindles by both visual and signal analysis measures. CONCLUSIONS: Most examples of hippocampal activity resembling spindles are probably physiologic, originating within the hippocampus or propagated from neighboring regions. However, in one subject, spindle activity and epileptiform discharges may have coincided, supporting experimental evidence that neurophysiological processes associated with spindle generation and NREM sleep contribute to the activation of epileptiform discharges. PMID- 10378740 TI - Minimal and asymptomatic chronic inflammatory demyelinating polyneuropathy. AB - OBJECTIVES: Show the chronic inflammatory demyelinating polyneuropathy (CIDP) is not only clinically heterogeneous but extremely variable in severity. METHODS: Three patients were referred for mild distal paresthesiae lasting more than 6 months and one for inguinal and thigh pain later ascribed to coxarthrosis. Strength was normal in all patients and tactile sensation reduced distally only in one. Tendon jerks were absent, except the knee jerks in one patient, reduced in lower limbs in two and normal in one. RESULTS: Electrophysiology showed a demyelinating neuropathy without motor conduction block. CSF protein content was increased in all patients. Nerve biopsies showed de-remyelination with varying degrees of axonal loss. Genetic studies excluded a demyelinating neuropathy associated with duplication or deletion of the 17p.11.2 segment. CONCLUSIONS: CIDP patients with pure sensory clinical presentation have been described but are generally more severely impaired. However, because of the mildness of symptoms and the unequivocal electrophysiological involvement of motor fibers, we think that in these cases the term minimal CIDP is more appropriate than sensory CIDP. These cases represent the most benign end of the CIDP spectrum. In our series minimal or even asymptomatic CIDP encompasses 8% of cases. PMID- 10378741 TI - Localization of the motor hand area using transcranial magnetic stimulation and functional magnetic resonance imaging. AB - OBJECTIVE: The anatomical location of the motor area of the hand may be revealed using functional magnetic resonance imaging (fMRI). The motor cortex representation of the intrinsic hand muscles consists of a knob-like structure. This is omega- or epsilon-shaped in the axial plane and hook-shaped in the sagittal plane. As this knob lies on the surface of the brain, it can be stimulated non-invasively by transcranial magnetic stimulation (TMS). It was the aim of our study to identify the hand knob using fMRI and to reveal if the anatomical hand knob corresponds to the hand area of the motor cortex, as identified by TMS, by means of a frameless MRI-based neuronavigation system. METHODS: Suprathreshold transcranial magnetic stimuli were applied over a grid on the left side of the scalp of 4 healthy volunteers. The motor evoked potentials (MEPs) were recorded from the contralateral small hand muscles, and the centers of gravity (CoG) of the MEPs were calculated. The exact anatomical localization of each point on the grid was determined using a frameless MRI-based neuronavigation system. In each subject, the hand area of the motor cortex was visualized using fMRI during sensorimotor activation achieved by clenching the right hand. RESULTS: In all 4 subjects, the activated precentral site in the fMRI and the CoG of the MEP of all investigated muscles lay within the predicted anatomical area, the so-called hand knob. This knob had the form of an omega in two subjects and an epsilon in the other two subjects. CONCLUSIONS: TMS is a reliable method for mapping the motor cortex. The CoG calculated from the motor output maps may be used as an accurate estimation of the location of the represented muscle in the motor cortex. PMID- 10378742 TI - Chronic inflammatory demyelinating polyneuropathy in diabetics: motor conductions are important in the differential diagnosis with diabetic polyneuropathy. AB - OBJECTIVE: It is important to recognize CIDP occurring in diabetics because, unlike diabetic polyneuropathy, it is treatable. The aim of this study was to find out whether there are clues which help to differentiate CIDP in diabetics from diabetic polyneuropathy. METHODS: We compared the electrophysiological and pathological findings of 7 diabetics, who developed a predominantly motor polyneuropathy with the features of CIDP, with a group of diabetics referred for symptomatic polyneuropathy. RESULTS: Of the 7 diabetics we believe developed CIDP, 6 met at least 3 and one patient two of the 4 electrophysiological criteria of demyelination. Of the 100 patients referred for diabetic polyneuropathy, only 4 fulfilled two criteria and none 3. Nerve biopsy findings were not helpful in differential diagnosis, as segmental demyelination and remyelination, onion bulbs and inflammatory infiltrates, which are the histologic features of CIDP, were also present in diabetic polyneuropathy. CONCLUSIONS: CIDP can be diagnosed in a diabetic patient when motor symptoms are predominant, are more severe than expected in diabetic polyneuropathy and 3 of the 4 electrophysiological criteria for demyelination are fulfilled. When only two criteria are met, we believe that a trial with one of the established treatments for CIDP may be helpful in confirming the diagnosis. PMID- 10378743 TI - Changes in motor evoked potentials to short-interval paired transcranial magnetic stimuli in multiple sclerosis. AB - OBJECTIVE: Paired transcranial magnetic stimuli (TMS) were applied in 8 multiple sclerosis (MS) patients with asymmetrical clinical signs and in 8 healthy controls to test the hypothesis that the circuits responsible for the generation and transmission of I-waves are abnormal in the former group METHODS: A figure-of 8 coil discharging through a Magstim 200/Bistim configuration delivered identical stimuli at an intensity 10% above the motor threshold of the relaxed first dorsal interosseous muscle. The interstimulus intervals (ISIs) used were varied in a pseudo-randomized fashion in steps of 0.2 ms between 1.0 and 5 ms. RESULTS: In 9 of 12 unilateral studies in the control group, a pattern of 3 peaks of increased motor evoked potential size was found at ISIs of 1.2-1.6 ms, 2.4-3.2 ms and 4.4 5.0 ms. A similar pattern was present in only 5 of 12 studies in the patients (Fisher's exact test, P = 0.1), while it was absent in all the 4 studies of the side with greater clinical involvement in patients (P = 0.01) CONCLUSION: Our results suggest that I-wave generation is more likely to be defective in MS than in normal subjects, that this defect resides in the cortex, and that it correlates with severity of physical signs. PMID- 10378744 TI - Spatial facilitation of motor evoked responses in monitoring during spinal surgery. AB - During spinal cord monitoring, motor responses in the tibialis anterior muscles were recorded on transcranial electrical stimulation of the motor cortex. In order to facilitate the responses, the cortical stimulus was preceded by a train of stimuli to the foot sole within the receptive field of the withdrawal reflex of the tibialis anterior muscle. This cutaneous input provides a spatial facilitation of the cortically elicited response. When the stimulus interval was 50-100 ms, large and reliable responses were seen in most cases. PMID- 10378745 TI - Reliability of surface electromyographic measurements. AB - OBJECTIVES: The aim of the study was to investigate short-term, intermediate-term and long-term reliability of surface electromyographic (EMG) measurements. METHODS: Eighteen healthy subjects performed 810 isometric knee extension tests. Reliability for maximum voluntary contraction (MVC) and 50% MVC was assessed with retest intervals of 3 min, 90 min and 6 weeks. Reliability for sustained contractions was assessed with retest intervals of 90 min and 6 weeks. EMG was recorded from the rectus femoris, vastus lateralis and vastus medialis muscles. The root mean square (RMS) and the median frequency (MF) parameters were extracted. At sustained contraction tasks, estimated linear regression values of both parameters were analyzed. Bland-Altman-plots, coefficient of repeatability, Pearson's coefficient of correlation and intra class correlation (ICC) procedures were applied to assess test-retest reliability. RESULTS: EMG recordings taken at short-term intervals were generally better reproducible than those of the longer term intervals. Moreover, 50% MVC EMG recordings demonstrated better reproducibility than 100% MVC measurements, and EMG recorded from the rectus femoris were more constant than that from the vastus lateralis or vastus medialis. The MF parameter recorded from the rectus femoris was the only reliable parameter of EMG fatigue change. CONCLUSION: In our set up, EMG measurement is best suited for clinical applications if submaximal MVC measurements are performed and signal is taken from rectus femoris muscle. PMID- 10378746 TI - F-chronodispersion in patients on thalidomide. AB - OBJECTIVES: To describe abnormalities of F-chronodispersion in patients treated with thalidomide. METHODS: We retrospectively studies F-wave latency, persistence and F-chronodispersion in 12 patients on thalidomide treatment and compared them with a control group of another 12 patients with similar dermatological conditions who did not receive thalidomide. Furthermore, we prospectively performed longitudinal neurophysiological studies in 4 patients before and during thalidomide treatment. RESULTS: Seven of 12 patients in the retrospective study had abnormal F-chronodispersion while this was normal in all patients of the control group (P = 0.014). All other neurophysiological parameters were similar in the two groups. Two of the thalidomide patients with abnormal F chronodispersion later developed sensory neuropathy. In all 4 patients in the prospective study though F-chronodispersion was normal before thalidomide it became markedly abnormal after exposure to this drug. CONCLUSIONS: Thalidomide may affect smaller diameter motor nerve fibres even before changes in sural sensory nerve action potentials. F-waves and F-chronodispersion should be routinely monitored in patients on thalidomide treatment. PMID- 10378747 TI - Assessment of temporal dispersion in motor nerves with normal conduction velocity. AB - Demyelinated nerves attenuate high-frequency components of propagating action potentials. In order to study if there is diagnostic use of this in motor nerves, the spectral energy above 49 Hz, amplitude, area, and duration of the compound muscle action potentials were measured; values after distal and proximal stimulation of posterior tibial nerves were compared. Normative data were collected in 48 control subjects. The same measurements were made in 20 patients with polyneuropathy and reduced motor nerve conduction velocity, in 21 patients with mild polyneuropathy but normal motor nerve conduction velocity, and in 8 patients with myasthenia gravis. Overall, high-frequency attenuation was closely correlated with amplitude decay (r = 0.63, P<10(-19)) and with increase of action potential duration (r = 0.34, P = 10(-5)). In the group of patients with normal NCV, high-frequency attenuation was abnormal in 9 (43%), amplitude decay was abnormal in two (10%), and area decay was abnormal in one (5%) patient. The action potential duration was normal in all of these patients. High-frequency attenuation was not influenced by stimulus intensity, thus it is not changed by conduction block, and it was not influenced by impaired neuromuscular transmission. Hence, high-frequency attenuation, both sensitively and specifically does indicate abnormal temporal dispersion. In conclusion, the simple measurement of high-frequency attenuation markedly improves detection and characterization of demyelination of human motor fibers. PMID- 10378748 TI - Demyelination and axonal degeneration in corpus callosum assessed by analysis of transcallosally mediated inhibition in multiple sclerosis. AB - OBJECTIVE: Following focal transcranial magnetic cortex stimulation (fTMS), inhibition of voluntary EMG activity in the ipsilateral first dorsal interosseus (FDI) muscle was studied, in order to assess the functional integrity of the corpus callosum in patients with multiple sclerosis (MS). METHODS AND RESULTS: Thirty-four patients suffering from definite MS and 12 healthy, age-matched normal subjects were examined. In mid-sagittal slices, 29 patients showed lesions within the truncus corporis callosi in T2-weighted MRI. In 20 patients, all areas (anterior, middle and posterior parts), in one both the anterior and posterior part, in 3 exclusively the anterior, in 4 the middle and in one the posterior area were affected. In 5 patients, lesions of corpus callosum were lacking. In normal subjects, fTMS elicited a transient inhibition (TI) of preactivated (50% of maximal force) isometric voluntary ipsilateral FDI muscle activity. Mean onset latencies of TI were 35.5+/-5.4 ms in right and 36.1+/-4.2 ms in left FDI. Mean duration of TI amounted to 23.0+/-8.4 ms for right and 24.6+/-8.4 ms for left FDI. In the MS group, TI latencies were significantly increased in 23 and TI durations in 16 cases, whereas a lack of TI was found in 5 patients bilaterally and in 6 unilaterally. In patients, mean onset latencies of TI were 40.4+/-13.8 ms in right and 43.3+/-14.4 ms in left FDI, TI duration amounted to 30.5+/-17.4 ms for right and 31.0+/-25.2 ms for left FDI. Increase of onset latencies and durations of TI were positively correlated with the summed area of lesions of corpus callosum in representative mid-sagittal MRI slices. Significant correlations between TI onset latencies and duration on the one hand, and central motor conduction latencies along corticospinal tracts (CML) on the other hand, were not found. CONCLUSION: The present investigation indicates that measurement of TI elicited by fTMS seems to be a sensitive method for an assessment of demyelination and axonal degeneration within corpus callosum in MS patients. PMID- 10378749 TI - Kinematics invariance in multi-directional complex movements in free space: effect of changing initial direction. AB - We investigated in normal human subjects the effect of changing the initial direction on the kinematic properties of figure '8' movement performed as fast as possible by the right arm extended in free space. To this end, the motion of the index finger was monitored by the ELITE system. The figure '8' movement was characterized by a complex tangential velocity profile (Vt) presenting 5 bell shaped components. It was found that the temporal segmentation following Vt was not significantly different, whatever the initial direction of the movement. The decomposition of Vt into different velocity profiles with respect to vertical (3 phases, Iy-IIIy) and horizontal (5 phases, Iz-Vz) directions showed a significant relationship between the amplitude and the maximal velocity for all the different phases (except the IIy phase), which demonstrated a good conservation of the Isochrony Principle. However, we showed that the transition between the clockwise and counter-clockwise loop (inflection point) induced greater variability in the vertical velocity profile than in the horizontal one. Moreover, some parameters such as the maximal velocity of Iy and the movement amplitude of the last phases (IIIy and Vz) showed significant changes depending on the initial direction. A highly significant positive correlation was observed between the instantaneous curvature and angular velocity. This was expressed by a power law similar to that previously describe for other types of movement. Furthermore, it was found that this covariation between geometrical and kinematic properties of the trajectory is not dependent on the initial direction of movement. In conclusion, these results support the idea that the fast execution in different directions of a figure '8' movement is mainly controlled by two types of invariant commands. The first one is reflected in the 2/3 power law between angular velocity and curvature and the second one is represented by a segmented tangential velocity profile. PMID- 10378750 TI - Waveform variation and size of sympathetic skin response: regional difference between the sole and palm recordings. AB - OBJECTIVE: The aim of this study was to investigate the regional difference in sympathetic skin response (SSR). The influence of SSR waveform from sole skin (S SSR) on latency, amplitude, and habituation was also studied. METHODS: Twenty SSRs were analyzed in 41 normal subjects. Waveforms were classified as either the P type, in which the positive component was larger than the negative one, or the N type, in which the negative component was larger the positive one. The occurrence patterns of these two waveform types were classified into three kinds, i.e. P, N, and M patterns. In the P or N pattern all the SSRs were of the P or N type. The M pattern had both P and N types during consecutive recordings. RESULTS: In the S-SSR, the P pattern had a higher amplitude and shorter latency than the N pattern, and habituation was most pronounced in the M pattern. These were compatible with previously reported findings in SSR from palm (P-SSR). The waveform patterns were not always consistent between P- and S-SSRs. The maximum S SSR and P-SSR were not simultaneously obtained in nearly half of the subjects. CONCLUSIONS: The size and waveforms of SSR were modified not only by the condition of the central component, but also the peripheral component of the reflex and the sweat gland. PMID- 10378751 TI - Normative data for onset VEPs to red-green and blue-yellow chromatic contrast. AB - OBJECTIVE: To better characterize the properties of chromatic VEPs to onset offset of red-green and blue-yellow equiluminant patterns, and establish normative values for a set of stimuli able to elicit robust and reliable responses, suitable for the clinical application. METHODS: Chromatic VEPs have been recorded (Oz lead) from 28 normal subjects (age range 20-53 years) in response to monocular presentation of both red-green and blue-yellow equiluminant sinusoidal gratings. Stimuli were generated by a Cambridge VSG/2 card and displayed on a Barco CCID monitor (14x14 deg field size). Spatial frequency, chromaticity, contrast and onset-offset duration were varied. RESULTS: For both red-green and blue-yellow equiluminant stimuli, robust responses have been obtained with gratings of 2 c/deg, presented in onset (300 ms) offset (700 ms) mode, at contrasts ranging from 90 to 6%. In all observers, the VEP waveform consisted mainly of a negative wave at stimulus onset, with a latency rapidly increasing with decreasing contrast. For both red-green and blue-yellow stimuli, the VEP contrast threshold coincided with the psychophysical threshold. CONCLUSIONS: The results complement previous studies aimed at characterizing the properties of chromatic VEPs. In addition, normative data are provided for a set of stimulus characteristics suitable for the clinical routine. PMID- 10378752 TI - Outcome studies. PMID- 10378753 TI - Executive function of children with extremely low birthweight: a case control study. AB - This study examines the executive function (EF) skills of extremely-low birthweight (ELBW) children at school compared with their peers. Thirty children with ELBW and 50 control children (both with a mean age of 62+/-4 months) were administered tests of EF including the Tower of Hanoi task, Finger Sequencing task, and Tapping Test. Children with ELBW, including those who scored more than 1 SD below the mean on the Peabody Picture Vocabulary Test-Revised, scored significantly lower than their peers on all executive tasks. There was limited correlation between EF and previous general quotient index scores obtained at routine assessment using the McCarthy Scales of General Ability at 4 years of age for the children with ELBW. Results suggest that children with ELBW are at risk for deficits in 'executive' behaviours including planning, sequencing, and inhibition which may have implications for later learning. PMID- 10378754 TI - Evidence of cognitive visual problems in children with hydrocephalus: a structured clinical history-taking strategy. AB - Damage to the occipital cortex in children can result in many complex disorders of cognitive visual function. A series of clinical questions, developed from the specific problems of a cohort of children with cortical visual impairment, was asked of the parents of 200 children with no history of cerebral pathology, aged 5 to 12 years. One hundred and ninety-two parents gave reliable consistent responses. The results show a progressive improvement in performance with age, culminating in few 11- and 12-year olds having frequent problems, apart from 8% having frequent difficulty with orientation in new surroundings and 2% having problems with simultaneous perception tasks. The parents of 52 children (aged 5 to 17 years) with shunted hydrocephalus were then asked the same set of questions. Evidence of cognitive visual problems was identified in 27 of these children of whom 16 manifested multiple difficulties. The disabilities identified by our study comprised problems with: shape recognition, simultaneous perception, perception of movement, colour perception, orientation, object recognition, and face recognition. The range, nature, and combinations of these disorders are presented in this paper. PMID- 10378755 TI - Survivors of neonatal extracorporeal membrane oxygenation at school age: unusual findings on intelligence testing. AB - Data are presented on 17 children who received extracorporeal membrane oxygenation (ECMO) in the neonatal period for persistent pulmonary hypertension (PPHN). These children are being followed as part of a larger program of follow up research on children who have been treated for PPHN with several treatment methods. On intelligence testing at ages 5 to 8 years, these 17 children had unusual patterns of results. A higher-than-predicted percentage of the ECMO survivors had discrepancies between their Verbal and Performance IQ and a much higher-than-predicted percentage had areas of unusual strength or weakness on their IQ subtest scores. Also, there was a significant correlation between the amount of time a child received ECMO and the child's Performance IQ: the longer the child received ECMO, the higher the Performance IQ. While findings of unusual weaknesses or deficits on intelligence testing at school age in children who have been severely ill in the neonatal period are not unusual, findings of high scores and areas of strength are not easily explained, particularly when these findings seem to relate to an invasive treatment like ECMO. Similar findings have been reported in two other small studies, which suggest that the impact of ECMO on the developing infant brain may not be purely detrimental. PMID- 10378756 TI - Spinal cord insults in the prenatal, perinatal, and neonatal periods. AB - We investigated the features of children with spinal cord insults (SCI) occurring in the pre-, peri-, and neonatal periods by sending 340 questionnaires to all paediatric neurologists, paediatric urologists, and neonatologists in the UK and Ireland. We requested information about timing, nature, and level of SCI in their patients; family and maternal history; pregnancy, delivery, and neonatal period; clinical presentation, imaging, laboratory studies, and outcome. Two-hundred and sixty-one questionnaires were returned with data on 58 patients with SCI. Seven out of the 58 children with SCI had pure dysraphic cord syndromes and were excluded. Fifty-one patients (33 males, 17 females, one unknown), born between 1972 and 1996, remained. Clinical presentations included severe respiratory failure (N=20; five of whom died neonatally) and hypotonia or weakness recognized either during the neonatal period (N=12) or after 28 days (N=10). Data on clinical presentation were not given in nine cases. Lesions were cervical (N=22) and thoraco-lumbar (N=29). SCI was ascribed to ischaemia (N=12), trauma (N=4), and other associated underlying conditions (N=11), whilst the aetiology was unknown in 24 cases. Mean gestational age (36.2 weeks) and birthweight (2.6 kg) were lower than previously reported with the lowest figures associated with thoraco-lumbar and ischaemic lesions. More males were affected by lesions than females and the incidence of preterm delivery, multiple pregnancy, breech presentation, forceps delivery, and caesarean delivery were higher than average. Forceps delivery was associated with cervical lesions. Outcome data were given in 47 children, nine of whom died either neonatally or within the first 20 months of life. Motor disability ranged from a complete recovery in one out of 40 to paraparesis in 26 out of 40, and tetraparesis in 13 out of 40 patients: 17 out of 39 were ambulant. Sphincter dysfunction was present in 22 out of 38 patients and scoliosis in 16 out of 37. Learning difficulties were present in 10 out of 39, behavioural problems in five out of 39 and seizures in four out of 39 patients. SCI in the pre-, peri-, and neonatal periods are rare but probably under diagnosed and are heterogeneous in aetiology, presentation, and outcome. Boys appear to be more susceptible than girls. PMID- 10378757 TI - Epilepsies of neonatal onset: seizure type and evolution. AB - Most neonatal seizures are occasional seizures and not true epilepsy. This study investigates seizure types of true neonatal epilepsies and their evolution with development. Seventy-five children with epilepsies of onset within 1 month of life, who were examined between 1970 and 1995, and whose seizure types could be confirmed with ictal EEG recordings, were studied. The patients were followed up for a minimum of 3 years and the evolution of epileptic syndromes was investigated. Sixty-three (84%) of 75 patients had partial seizures, while nine had generalized seizures, and only three had both generalized and partial seizures. Twenty-three of 24 neonates with benign familial or non-familial neonatal convulsions presented with partial seizures; these syndromes should not necessarily be categorized into generalized epilepsy as they are in the present International Classification. Age-dependent changes were a common feature of symptomatic neonatal epilepsies. Eighteen (41%) of 44 patients with symptomatic epilepsies of neonatal onset developed West syndrome in infancy. Fifteen (83%) of these 18 patients presented with symptomatic localization-related epilepsy in the neonatal period. In seven of these 15 patients, West syndrome was followed by localization-related epilepsy. Symptomatic localization-related epilepsy with transient West syndrome in infancy is another type of age-dependent epileptic syndrome. PMID- 10378758 TI - A population-based approach to the investigation of osteopenia in Rett syndrome. AB - This study compares bone mass in a national sample of girls with Rett syndrome (RS) with a sample of control children. The Australian RS Database was the source of cases for this population-based study. Hand radiographs were available from 101 of 137 subjects (74% of the known Australian population of girls with RS aged < or = 20 years). Control radiographs matched for age, sex, and laterality were obtained from hospital radiology departments. A measure of cortical thickness was made from the difference between the outer diameter and the medullary space in the second metacarpal bone. A mean z-score value for cortical thickness and percentage cortical area for each individual was calculated. The mean cortical thickness (z score) for girls with RS was -1.94 compared with -0.38 for control children (P<0.001). In girls with RS, the mean cortical thickness decreased with age (P<0.001). In girls who were taking epilepsy medication it was -2.21 compared with -1.23 in those not taking epilepsy medication (P<0.001). There was no evidence of a beneficial effect of increased calcium intake on cortical thickness. A similar pattern was obtained when percentage cortical area was estimated. In multivariate analysis, increasing age and use of anticonvulsant medication were associated with decreased cortical thickness and only use of anticonvulsant medication with decreased percentage cortical area. Fractures had occurred in one-third of cases and it was estimated that just over 40% of girls would sustain a fracture by the age of 15 years. Girls with RS may be at increased risk of fractures and their bone quality compromised as determined by cortical thickness and percentage cortical area measurements from the second metacarpal. PMID- 10378759 TI - Respiratory tract infections due to direct and reflux aspiration in children with severe neurodisability. AB - Lower respiratory tract infections in children with severe neurodisability are usually caused by aspiration of stomach contents from gastroesophageal reflux (GOR) or direct aspiration (DA) of food due to oral and pharyngeal motor problems. To determine the contributions and interactions of GOR and DA, oesophageal 24-hour pH monitoring and feeding videofluoroscopy were performed in 34 children (age range 7 months to 16 years, mean 7 years) who had severe physical and learning disabilities and who were slow feeders. Subjects were divided into three groups according to the frequency of their respiratory tract infections. Subjects in group 1 had no respiratory tract infections (N=10); five had GOR and none had DA. Subjects in group 2 had minor respiratory tract infections but had not received more than one course of antibiotics for this in the previous year (N=8); two had GOR alone, four had DA alone, and two had neither. All subjects in group 3 had recurrent respiratory tract infections (N=16); one had GOR alone, seven had DA alone, and eight had both GOR and DA. This study suggests that oral and pharyngeal motor problems are the major cause of respiratory tract infection in children with severe neurodisability. These problems lead to DA and, if GOR is present, to the aspiration of stomach contents. Those children with both DA and GOR are more likely to have severe respiratory tract infections which may lead to gastrostomy feeding (together with fundoplication). GOR without sufficient oral and pharyngeal motor problems to cause DA is less likely to cause respiratory tract infection in children with severe neurodisability. PMID- 10378760 TI - Speech discrimination and phonological working memory in children with ADHD. AB - This study examined phonological working memory and speech discrimination among children with attention-deficit-hyperactivity disorder (ADHD) with and without motor problems. Forty-one children were assigned to three groups; children with ADHD (N=9), children with ADHD plus developmental coordination disorder (ADHD+, N=13), and age-matched control children (N=19). The subjects' ability to classify stimulus pairs was examined in two experiments. The first experiment required subjects to discriminate pairs of monosyllabic stimuli with contrasting consonants to test speech discrimination without using a working-memory load. In the second protocol, subjects were exposed to two- to five-syllabic non-word pairs with contrasting vowels in order to test speech discrimination with a working-memory load. The subjects classified the pairs as being either the same or different in both experiments. No significant differences were found between the subject groups in the discrimination task with monosyllables. When exposed to the two- to five-syllabic stimuli, the ADHD+ group scored significantly lower than both other groups. This was attributed to a higher sensitivity to working memory load. Some possible explanations of this effect are discussed. PMID- 10378761 TI - Respiratory depression in children receiving diazepam for acute seizures: a prospective study. AB - The aim of this study was to determine the incidence of respiratory depression following the use of diazepam in children presenting with seizures. All children presenting with seizures to a children's A & E department over a period of 9 months were studied prospectively. Respiratory depression was defined as a fall in respiratory rate or oxygen saturation, or apnoea resulting in ventilation or resuscitation with bag-and-mask oxygen. There were 130 patient episodes involving 97 children who received treatment for their seizures before admission and/or in the A & E department. Administration of diazepam resulted in 122 patient episodes. The route of administration was rectal in 91 episodes, intravenous in 12 episodes, and both rectal and intravenous in 19 episodes. Eleven children had respiratory depression in relation to diazepam administration. Eight of these children required ventilation. The overall incidence of respiratory depression following the use of diazepam was 9%. The incidence of respiratory depression following diazepam given intravenously or rectally is high. The use of diazepam as first-line therapy for children with acute seizures needs to be reviewed. PMID- 10378762 TI - Ictal hemiparesis. AB - Two subjects with ictal hemiparesis are described. Both children presented with evolving paresis associated with seizure activity. Structural neuroimaging remained consistently normal, although EEG demonstrated slow-wave activity, and SPECT scanning in one child showed perfusion asymmetry. Both children had resolution of the hemiparesis when seizure activity was adequately controlled. The historically proposed pathophysiology of ictal hemiparesis is that of inhibition of the somatosensory and motor areas of the cortex. The presence of an evolving hemiparesis and seizure activity associated with normal neuroimaging should prompt consideration of ictal hemiparesis. Confirmation of this rare diagnosis can only be made when seizure control leads to resolution of the paresis. PMID- 10378763 TI - Sleep disorders in visually impaired children. PMID- 10378764 TI - Ataxia of parietal lobe origin. PMID- 10378765 TI - Prevalence of Tourette syndrome. PMID- 10378766 TI - Identification of PTEN-related sequences in glioma cells and in non-neoplastic cell lines. AB - The PTEN gene, which encodes a tumor suppressor with phosphatase activity, is located on chromosome 10q23 and is mutated in different tumors, including glioblastomas (GBM). We found evidence for a PTEN-related sequence (PTEN-rs) on genomic DNA of GBM and non-neoplastic cells. PTEN-rs does not contain introns and presents several conserved missense mutations, including a T to G transversion at the initiation codon. Rsa I digestion may help to identify this putative PTEN pseudogene which, according to RT-PCR analysis on glioma, fibroblast, brain and lung cells, does not appear to be transcribed. PMID- 10378767 TI - Enhanced efficacy of 1-methyl-3-propyl-7-butylxanthine on the antitumor activity of doxorubicin against doxorubicin-resistant P388 leukemia. AB - The effects of 1-methyl-3-propyl-7-butylxanthine (MPBX), a xanthine derivative, on doxorubicin (DOX)-induced antitumor activity against DOX-sensitive P388 leukemia (P388) and DOX-resistant P388 leukemia (P388/DOX) have been examined. In P388-bearing mice, the combination of MPBX with DOX increased the antitumor activity of DOX 1.6-fold. In contrast, in P388/DOX-bearing mice, DOX alone did not decrease the tumor weight, whereas in combination with MPBX it significantly decreased the tumor weight in the control group by 50%. The increase in DOX induced antitumor activity caused by MPBX was correlated with the DOX concentration in the tumors. The DOX concentration in the tumors of P388- and P388/DOX-bearing mice in the MPBX combination group increased by 1.3-fold and 2.2 fold, respectively, compared to the level in the DOX-alone group. On the other hand, there was no increase in the DOX concentration in the heart or liver in both types of tumor-bearing mice treated with MPBX. In vitro, the facilitated DOX influx and suppressed efflux by MPBX in both types of tumor cells were similar, suggesting that MPBX acts on the same site in both types of cells. P388/DOX overexpressed P-glycoprotein, i.e. the inhibitory order of DOX efflux caused by the inhibitor of P-glycoprotein was P388 < P388/DOX. However, the effect of MPBX was P388 > P388/DOX. Therefore, we expect that the site of attack by MPBX is not P-glycoprotein. PMID- 10378768 TI - An enhanced active efflux of CPT-11 and SN-38 in cisplatin-resistant human KB carcinoma cells. AB - Cisplatin-resistant KCP-4 cells were 12.4- and 31.6-fold more resistant to CPT-11 and SN-38 than parental KB-3-1 cells, respectively. We studied the mechanism of cross-resistance to CPT-11 and SN-38. Our previous study showed that multidrug resistance protein (MRP), canalicular multispecific organic anion transporter (cMOAT) and P-glycoprotein (P-gp) were not expressed in KCP-4 cells (Chen, Z.-S. et al., Exp. Cell Res., 240 (1998) 312-320, and Chuman, Y. et al., Biochem. Biophys. Res. Commun., 226 (1996) 158-165). The accumulation of both CPT-11 and SN-38 in KCP-4 cells was lower than that in KB-3-1 cells. The ATP-dependent efflux of CPT-11 and SN-38 from KCP-4 cells was enhanced compared with that from KB-3-1 cells. DNA topoisomerase (topo) I expression, topo I activity, topo I mediated cleavable complex, and the sensitivity to SN-38 of DNA topo I in KCP-4 were similar to those in KB-3-1 cells. Furthermore, the conversion of CPT-11 to SN-38 in the two cell lines was also similar. The transport of LTC4 in KCP-4 membrane vesicles was competitively inhibited by bis-(glutathionato)-platinum (II) (GS-Pt), CPT-11 and SN-38. These findings suggested that an unknown transporter distinct from P-gp, MRP or cMOAT is expressed in KCP-4 cells and transports CPT-11 and SN-38. PMID- 10378769 TI - Prevention of spontaneous and chemically induced carcinogenesis using activated carbon fiber adsorbent. II. Inhibitory effect of the activated carbon fiber adsorbent 'Aqualen' on N-methyl-N'-nitro-N-nitrosoguanidine-induced gastric carcinogenesis in rats. AB - Two-month-old female LIO rats were given N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) with tap water in a concentration of 100 mg/l for 12 months (groups 1 and 2) or were kept without the carcinogen treatment (groups 3 and 4). From the first day of exposure to MNNG rats from groups 2 and 3 were given activated carbon fiber adsorbent Aqualen in their diet five times per week together with lab chow in a daily dose of 100 mg/kg of body weight. The experiment was finalized 16 months after first exposure to the carcinogen. The total stomach adenocarcinoma incidence was 43% in group 1 and 39% in group 2, whereas invasive stomach adenocarcinomas occurred in 36% and 8% of rats from groups 1 and 2, respectively (P < 0.05). Tumors other then stomach sites (duodenum and liver) only developed in rats from group 1 (29%). No lesions were observed in rats exposed to Aqualen without MNNG. Thus, our results demonstrate the inhibitory effect of the activated carbon fiber adsorbent Aqualen on stomach carcinogenesis in rats. PMID- 10378770 TI - Prevention of spontaneous and chemically induced carcinogenesis using activated carbon fiber adsorbent. III. Inhibitory effect of the activated carbon fiber adsorbent 'Aqualen' on 1,2-dimethylhydrazine-induced intestinal carcinogenesis in rats. AB - Two-month-old outbred female LIO rats were exposed weekly to 15 (experiment I, groups 1, 2 and 3) or to 5 (experiment II, groups 4, 5 and 6) subcutaneous injections of 1,2-dimethylhydrazine (DMH) at a single dose of 21 mg/kg of body weight. From the day of the first injection of the carcinogen, the rats from groups 2, 3, 5 and 6 were given Aqualen in their diet. In both experiments rats were fed Aqualen five times per week together with lab chow at the daily dose of 0.1 g/kg (groups 2 and 5) or 1.0 g/kg (groups 3 and 6) of body weight. Additionally, other rats were not exposed to the carcinogen and served as an intact control (group 7) or were given Aqualen with the diet at the daily dose of 0.1 g/kg (group 8) or 1.0 g/kg (group 9). These experiments were finalized 6 months after the first injection of DMH. In experiments I and II, the majority of tumors were localized in the descending colon. Tumors of the small intestines developed only in rats from experiment I. The total incidence of colon tumors as well as tumors in different parts of the colon and the mean number of tumors per rat were much higher in rats from all groups in experiment I than in the rats from experiment II. In experiment I supplementation of Aqualen to the diet was followed by a decrease in the incidence of tumors in the ascending colon and by a decrease in the number of tumors per rat in both ascending and descending colons regardless of the dose of the enterosorbent. In experiment II the effect of Aqualen was stronger than in experiment I -- the enterosorbent decreased both the tumor incidence and the multiplicity in the total colon, its ascending and descending parts and in the rectum. In experiments I and II the percentage of small colon tumors among rats exposed to Aqualen (groups 2, 3, 5 and 6) was higher than that of the controls (groups 1 and 4). Most of detected intestinal tumors were classified as adenocarcinomas. The level of tumor differentiation was higher in rats exposed to Aqualen. There were no pathological changes observed in rats exposed to Aqualen without DMH. Carcinogen treatment resulted in an increase of serum glucose and cholesterol levels whereas Aqualen normalized these changes. Thus, our results demonstrate the inhibitory effect of activated carbon fiber adsorbent Aqualen on intestinal carcinogenesis in rats. PMID- 10378771 TI - Effects of melatonin on N-nitroso-N-methylurea-induced carcinogenesis in rats and mutagenesis in vitro (Ames test and COMET assay). AB - The effect of melatonin, an indole hormone of the pineal gland, on the initiation of N-nitroso-N-methylurea (NMU)-induced carcinogenesis in rats and mutagenesis in vitro has been investigated. Two-month-old female LIO rats (groups 1 and 2) were exposed to a single injection of NMU (50 mg/kg of body weight, i.v.). Rats from group 2 were given melatonin orally (20 mg/l) from 18:00 to 09:00 h over 3 days (2 days before and 1 day after NMU injection). Animals from group 1 (control) were administered the solvent (ethanol/water, 1:1000). Rats were followed up to natural death or were sacrificed when moribund. Tumors developed both in rats treated with NMU alone (50.0%) and in rats exposed to NMU plus melatonin (34.8%). The percentage of malignant tumor-bearing rats in group 2 (21.7%) was lower (P < 0.02) than that in the other group (41.7%). Melatonin also decreased the multiplicity of malignant tumors 1.3-fold and reduced the incidence of malignancies in some organs. Two in vitro tests were used for mutagenesis studies: the Ames test (strains TA 100 and TA 102 of Salmonella typhimurium) and the Single Cell Gel Electrophoresis assay (SCGE assay or COMET assay) performed on CHOK1 cells. Melatonin itself revealed no genotoxic effect in either of the tests. No protective action of melatonin (at doses of up to 2 micromol/plate) towards NMU was found in the Ames test. In contrast, in the SCGE assay a slight, but statistically significant (P < 0.001), dose-related anticlastogenic effect of melatonin (10(-10)-10(-7) M) was observed. Thus, our data indicate that melatonin may act as an anti-initiating hormone in NMU-induced carcinogenesis and possess anticlastogenic activity towards NMU in CHOK1 cells. PMID- 10378772 TI - Detection of cerebroside sulfotransferase mRNA in human gastric mucosa and adenocarcinoma. AB - Sulfatide is a major acidic glycolipid in human gastric mucosa, and its sulfation is catalyzed by cerebroside sulfotransferase (CST). To investigate the expression of the CST gene in human gastric cancer, a reverse transcription PCR method was developed with the use of endoscopic bioptic specimens. By this method, we examined the CST mRNA expression in 11 cases of gastric cancer, and in all the cases we detected various levels of the expression both in cancer tissues and in uninvolved adjacent tissues. The present assay method was suggested to be useful in the detection of CST mRNA from a limited amount of bioptic samples. PMID- 10378773 TI - Heme oxygenase-1 expression in oral squamous cell carcinoma as involved in lymph node metastasis. AB - Thirty-eight oral squamous cell carcinomas (SCCs) were semi-quantitatively analyzed by immunohistochemical staining, and the relation between heme oxygenase 1 (HO-1) expression and the clinical status were correlated. High immunostaining of HO-1 was detected in lymph node metastasis negative groups (P = 0.0018) and in well-differentiated SCCs (P = 0.0016). There were no significant correlations between heme oxygenase-1 expression and other factors, such as size of the tumor, staging, age and sex. These findings further support the proposition that high heme oxygenase-1 expression in oral SCCs can be useful in identifying patients at low risk of lymph node metastasis. PMID- 10378774 TI - Direct evidence for the formation of deoxyribonucleotide adducts from carcinogenic N-nitroso-N-methylaniline revealed by the 32P-postlabeling technique. AB - N-Nitroso-N-methylaniline (NMA) is an esophageal carcinogen in the rat. NMA forms a benzenediazonium ion (BDI) during microsomal cytochrome P-450 2B1 (CYP2B1) catalyzed metabolism. Using the nuclease P1-enhanced version of the 32P postlabeling assay we investigated the formation of adducts by NMA with deoxyadenosine 3'-monophosphate (dAp) and deoxyguanosine 3'-monophosphate (dGp). 32P-postlabeling analysis of dAp and dGp, which were modified by NMA activated with microsomes of rats pretreated with phenobarbital (PB), and directly labeled resulted in each case in the appearance of one single adduct spot. Quantitative analysis of adducts revealed that the extent of dGp modification by activated NMA was more than 23 times greater than the extent of modification of dAp. The results suggest strongly that BDI, derived from NMA by CYP2B1 present in PB microsomes, participates in the formation of dAp and dGp adducts. PMID- 10378775 TI - A comparison of the NK cell cytotoxicity with effects of TNF-alpha against K-562 cells, determined by LDH release assay. AB - Effects of r h TNF-alpha as a single cytotoxic mediator against K-562 cells was examined by LDH release and compared with NK cell cytotoxicity. The mean values of the percentage of LDH release (x = 6.25 +/- 3.68%, for ten individual experiments) from K-562 cells cultured for 2 h with r h TNF-alpha 100 U/ml of culture medium did not give significant difference in comparison with mean values of percentage LDH release (x = 6.43 +/- 2.97%, for 37 individual experiments) from K-562 cells which were cultured without r h TNF-alpha (Student's t-test, P > 0.05). The results also showed, that in the presence of increasing concentrations of r h TNF-alpha there was no significant increase of LDH release through the cell membrane in these short term incubations. However, significant difference in LDH release from K-562 cells was found after 6 h between cultures treated for 30 min with or without r h TNF-alpha (Mann-Whitney test, P < 0.05). Since TNF-alpha alone shows a lower degree of K-562 cell membrane damage than NK effectors, this suggested that TNF-alpha is neither an only nor a major mediator of cell destruction, based on determination of LDH release. PMID- 10378776 TI - Multiple cytochrome P-450 subfamilies are co-induced with P-glycoprotein by both phenothiazine and 2-acetylaminofluorene in rats. AB - We studied the effects of two P-glycoprotein (P-gp) inducers, 2 acetylaminofluorene (2-AAF) and phenothiazine (PTZ), administered intraperitoneally, on the activities and content of hepatic cytochrome P-450 (CYP) subfamilies in hepatic microsomes of Sprague-Dawley rats. After 4-day administration of 2-AAF or PTZ, the P-gp content was increased. The total CYP content after PTZ treatment was significantly increased compared with that of controls. The CYP1A, CYP2B and CYP3A2 contents were induced, while the CYP2C6, CYP2C11 and CYP2E1 contents remained unaffected. A marked increase in CYP1A1 was found after administration of each compound. Ethoxyresorufin O-deethylase, pentoxyresorufin O-deethylase, and testosterone 6beta hydroxylation activities showed a significant increase after both 2-AAF and PTZ treatments. In particular, ethoxyresorufin O-deethylase exhibited more than ten times greater activity than that of the controls after the treatments. These results suggest that P-gp inducers affect several CYP subfamilies in addition to CYP3A, which is reported to be up-regulated coordinately with P-gp by a CYP3A inducer. PMID- 10378777 TI - Dose-dependent induction of aberrant crypt foci in the colons but no neoplastic lesions in the livers of heterozygous p53-deficient mice treated with low dose 2 amino-3-methylimidazo [4,5-f]quinoline. AB - 2-Amino-3-methylimidazo[4,5-f]quinoline (IQ) is a food derived heterocyclic amine which induces aberrant crypt foci (ACF) and tumors in the livers in mice. However, most previous studies of carcinogenicity were carried out with high dose treatments, so the practical risk associated with the low dose exposure is unclear. We, therefore, assessed whether low dose IQ causes ACF formation in the colons of mice constitutively hemizygous for functional p53. Simultaneously, we screened for development of preneoplastic foci in the liver. A total of 60 heterozygous p53-deficient mice as well as 60 wild-type mice were divided into five groups and administered IQ in the diet at concentrations of 50, 10, 2, 0.4 and 0 ppm until the end of the experiment. ACF were detected in the 50, 10 and 2 ppm-treated groups and the numbers of those comprising one aberrant crypt (AC) in p53-deficient mice treated with 10 or 2 ppm were significantly increased, compared to counterpart wild-type values. A dose-dependent increase of ACF was also observed in transgenic mice groups but no large ACF developed. In spite of extensive examination, no preneoplastic foci could be detected in either transgenic or wild-type mice. The results suggested that germline p53 deficiency may slightly enhance the development of ACF in colons but not in the liver. The fact that no ACF were detected in the lowest, 0.4 ppm, treated groups may imply a practical non-effective level of IQ for tumor induction. PMID- 10378778 TI - Anti-tumor-promoting effects of 8-substituted 7-methoxycoumarins on Epstein-Barr virus activation assay. AB - In a search for anti-tumor-promoting agents, we carried out a primary screening of twenty-nine 8-substituted and four 6-substituted derivatives of 7 methoxycoumarins isolated from plants of the Murraya and/or Citrus species (Rutaceae), examining their possible inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by 12-O-tetradecanoylphorbol-13-acetate (TPA) in Raji cells. This investigation indicated that the prenyl (3-methyl-2 butenyl) or 2-hydroxy-3-methylbutyl (or butenyl) unit as an isoprenoid moiety at C-8 on the 7-methoxycoumarin nucleus plays an important role in the anti-tumor promoting activity. Some of the 8-substituted 7-methoxycoumarins isolated from Murraya species, murrangatin (7), minumicrolin (10) and chloticol (18), were found to significantly inhibit EBV-EA activation, and preserved the high viability of Raji cells, suggesting that 7, 10 and 18 might be valuable anti tumor-promoting agents. PMID- 10378779 TI - Radiation-induced tumorigenesis of mammary glands in pituitary transplanted rats ovariectomized before onset of estrous cycle. AB - The role of prolactin in the initiation of mammary tumorigenesis by radiation was evaluated in ovarian hormone-free rats. Rats were bilaterally ovariectomized at 23 days of age, and then, at 2.5 months of age, two pituitaries obtained from mature rats of the same strain were transplanted underneath the kidney capsule as a means of increasing the serum prolactin level to provide stimulation of development of mammary glands. After 2 weeks, the ovariectomized rats with ectopic pituitary glands were exposed to whole body irradiation of 2.6 Gy of gamma-rays from a 60Co source and then treated with diethylstilbestrol as a tumor promoter. For the control, ovariectomized rats without ectopic pituitary glands were exposed and treated in the same way as the experimental group. A significant increase of serum prolactin level was observed at the time of irradiation by the pituitary transplanted rats, and intense immunohistochemical reaction with a specific anti-prolactin antiserum was detected in the ectopic pituitary glands. Also, mammary glands in the pituitary transplanted rats, ovariectomized before puberty, showed lactiferous ducts without alveolar buds at the time of tumor initiation. The pituitary transplanted rats showed a significantly increased incidence of adenocarcinoma and fibroadenoma compared with the control. Many of the mammary tumors induced in the pituitary transplanted rats given radiation were estrogen receptor (ER)(+) progesterone receptor (PgR)(+) and ER(+)PgR(-) tumors, whereas ER(-)PgR(-) tumors were mainly obtained in the control rats. In the experimental group, many of the fibroadenomas had low concentrations of ER and no PgR, while the adenocarcinomas had moderate concentrations of ER and high PgR. These results suggest that hypersecretion of prolactin from the pituitary transplants developed lactiferous ducts and accelerated the tumorigenesis of mammary glands initiated by radiation in the absence of synergism with ovarian hormones. PMID- 10378780 TI - Flow cytometric DNA analysis and chromosomal aberrations in malignant glioblastomas. AB - In this study we combined flow cytometry with fluorescence in situ hybridization to detect numerical aberrations in chromosomes. Fifty-nine human malignant gliomas were examined by flow cytometry for DNA-content and cell cycle analysis and for numerical aberrations of chromosome 1 by in situ hybridization using a chromosome specific centromere probe. Of the gliomas analysed, 42% were diploid and 58% showed aneuploid tumour cell populations. The DNA index was heterogeneous ranging from 1.0 to 2.3. The S-phase analysis showed proliferation activity from a very low range of 0.7% up to 17.0%. In general, diploid gliomas exhibited a lower S-phase activity than aneuploid gliomas. Of the aneuploid gliomas, 15% showed a peridiploid pattern with a DNA index mean of 1.1. In these peridiploid tumours a trisomy of chromosome 1 could be detected by fluorescence in situ hybridization (FISH). The frequency of trisomic chromosome 1 in malignant gliomas reflects a very slight increase in DNA index from diploid to peridiploid (DNA index 1.1). Comparison of chromosome numbers and DNA content gave good correlation. Also important, the results reflects the cell cycle, specifically the extent of S-phase activity. In general, cell proliferation of diploid and peridiploid gliomas is much less than in higher aneuploid gliomas. The analysis of DNA content may thus yield results with respect to the biological behaviour of tumours in general. PMID- 10378781 TI - Evaluation of toxicity of beta-tethymustine, a new anticancer compound, in mice. AB - The toxicity of beta-tethymustine, a potential anticancer compound 1 ((Cancer Lett., 119 (1997) 7-12) was assessed in normal as well as in Ehrlich ascites carcinoma (EAC), Sarcoma-180 (S-180) and Dalton' s Lymphoma (DL) tumour-bearing Swiss male mice by measuring drug-induced changes in haematological parameters, femoral bone marrow cellularity and splenic cellularity on days 9, 15 and 21 following drug treatment at the optimum dose of 8.0 mg/kg body weight from days 1 to 7. Detailed studies were also made by noting sequential changes in the above parameters in normal and EAC-bearing mice on days 12 and 18, respectively. The results indicate that the compound did not adversely affect haematopoiesis as it was observed that no significant decrease in haematological parameters and femoral marrow cellularity occurred in treated groups. Initial hyposplenic activity was, however, noted in EAC and normal treated groups on day 9 which soon reached normal count within 7-10 days after termination of drug therapy. Drug induced hepatotoxicity and nephrotoxicity were also sequentially evaluated in normal and tumour-bearing mice on days 9, 15 and 21 but no such toxicities were detected. Also, body weight, skin and hair texture, and behavioural pattern (food and water intake and activity) did not reflect any toxic reaction in host mice at this optimum dose. PMID- 10378782 TI - Human erythrocyte acetylcholinesterase inhibition by cis-diamminediaquaplatinum (II): a novel kinetic approach. AB - The present work addresses the analyses of some novel kinetic parameters (k(t), K(v), t50, K(ir), t(c), m(c), IC50, IC99 and Ki) of human erythrocyte membrane bound acetylcholinesterase (AChE, EC 3.1.1.7) inhibition by cis diamminediaquaplatinum II (PDC). PDC is under a clinical trial for use as an antineoplastic drug. The authors recently reported that PDC and cisplatin have the ability to inhibit AChE activity in vitro. Therefore this study was designed to determine the estimation of time constant (k(t)), velocity constant (K(v)), 50% inhibition time (t50), inhibition rate constant (K(ir)), transition concentration (t(c)), meeting concentration (m(c)), 50% inhibition (IC50), 99% inhibition (IC99) and inhibition constant (Ki) by novel methods. The details are described in the text. PMID- 10378783 TI - Butyrolactone I induces cyclin B1 and causes G2/M arrest and skipping of mitosis in human prostate cell lines. AB - Several naturally occurring cyclin-dependent kinase (CDK) inhibitors have been isolated from different lower organisms. In this report, we examined the effect of one of the CDK inhibitors, butyrolactone I (BL), on the expression of cyclins D2, A and B1 in three human prostatic cancer cell lines (DU145, PC-3, LNCaP) using two colored flow cytometric analysis. The percentage of DU145 cells in the 4C phase of the cell cycle were increased significantly at both 70 microM and 100 microM BL. Furthermore, an additional 8C peak was observed which had double the DNA content of the 4C phase at these concentrations of BL. The appearance of the 8C peak increased gradually and was more evident in DU145 and PC-3 than LNCaP. Cells in the 8C peak had either two nuclei or abnormal nuclei as observed by Papanicolaou stain. BL also increased the amount of cyclin B1 positive cells in the 4C phase. This increase was apparent on day 1 and returned to normal by day 3. Since BL selectively inhibits cyclin-dependent kinase, cyclin B1 might accumulate without being degraded. Other cyclins were not significantly changed by BL. The data demonstrate that BL inhibited Cdc2 of unsynchronized cultured prostate cancer cells, and interrupted the cell cycle progression toward cell division. The BL inhibition of Cdc2 led to the accumulation of cells in the 4C phase without mitosis resulting in an accumulation of cyclin B1. The appearance of cells in the 8C phase may be due to the progression of cells in the 4C phase through the cell cycle skipping mitosis. Cyclin B1 decreased in correlation with the progression through a new cell cycle. These results suggest that BL does not cause a complete arrest of the cell cycle in G2/M but that BL occasionally allows for the skipping of mitosis and subsequent progression through the cell cycle to occur. PMID- 10378784 TI - Suppressive effects of alpha-Hederin on 2,3,7,8-tetrachlorodibenzo-p-dioxin mediated murine Cyp1a-1 expression in the mouse hepatoma Hepa-1c1c7 cells. AB - Cultured mouse hepatoma cell line Hepa-1c1c7 cells were treated with alpha Hederin to assess the role of alpha-Hederin in the process of Cyp1a-1 induction. Treatment of Hepa-1c1c7 cultures with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induced Cyp1a-1, as indicated by analysis of 7-ethoxyresorufin O-deethylation (EROD) activity and Cyp1a-1 protein. When alpha-Hederin and TCDD were both added to cultures, TCDD-inducible EROD activity was greatly suppressed by alpha-Hederin in a dose-dependent manner. TCDD-induced Cyp1a-1 protein and mRNA levels were markedly reduced in the concomitant treatment of TCDD and alpha-Hederin consistent with EROD activity. Electrophoretic mobility shift assay using nuclear extraction of cells revealed that alpha-Hederin reduced transformation of the Ah receptor to a form capable of specifically binding to an oligonucleotide containing a dioxin-response element (DRE) sequence of the Cyp1a-1 gene. These results suggest that the suppressive effect of alpha-Hederin on TCDD-induced Cyp1a-1 gene expression in Hepa-1c1c7 cells might be an antagonist of the DNA binding potential of a nuclear Ah receptor. PMID- 10378785 TI - The association between nm23-H1 expression and survival in patients with esophageal squamous cell carcinoma. AB - The nm23 gene is a potential metastasis suppressor gene originally identified using a murine melanoma cell line. The expression of nm23-H1 protein was examined immunohistochemically in 50 eligible patients with esophageal squamous cell carcinoma (ESCC). The expression was not correlated with other prognostic factors including lymph node metastases; however, overall survival rates of nm23-H1 negative patients were significantly shorter than those of nm23-H1-positive patients (P < 0.05). Furthermore, reduced expression of nm23-H1 was associated with shorter overall survival in patients with involved lymph nodes (P < 0.01), but not in patients without involved lymph nodes. These data support the conclusion that reduced expression of nm23-H1 may be associated with poor prognosis of ESCC patients, suggesting the value of nm23-H1 expression as a prognostic marker for ESCC patients, especially ESCC patients with involved lymph nodes. PMID- 10378786 TI - Selenium-enriched Agaricus bisporus mushrooms suppress 7,12 dimethlybenz[a]anthracene bioactivation in mammary tissue. AB - The present studies compared dietary Se (1.0 microg/g) when provided as either fortified Agaricus bisporus mushrooms, or sodium selenite on the in vivo metabolism of 7,12-dimethylbenz(a)anthracene (DMBA). Dietary addition of Se unenriched A. bisporus mushrooms at 2% did not alter the occurrence of DMBA induced DNA adducts or the activity of glutathione S-transferase (GST). However, the addition of Se as enriched mushrooms, or as selenite, significantly increased both liver and mammary GST activity. Providing sodium selenite, or enriched mushrooms also significantly reduced total and anti-3,4-dihydrodiol-1,2-epoxide deoxyguanosine adducts compared to feeding the basal diet (P < 0.05). These investigations provide evidence that Se enriched mushrooms can be used as an effective method to retard chemically induced tumors. PMID- 10378787 TI - Progressive changes of the nuclear matrix protein composition in diethylnitrosamine-induced rat hepatocarcinogenesis. AB - The nuclear matrix proteins (NMPs) consist of common and tissue-specific nuclear proteins, which can be altered by extracellular environments such as radiation, chemicals and virus infection. Thus, NMP profiles were analyzed in diethylnitrosamine (DEN)-treated rat liver. Male F344 rats (6 weeks old) were treated continuously with DEN (40 ppm) in drinking water. All animals were sacrificed at 10, 20 or 30 weeks during the experiment. The profiles of NMPs and cytoskeletal proteins (CSPs) progressively changed in their number and amount in DEN-treated rats. Four peptides increased in their relative amounts, while three decreased in the 10- and 20-week groups. Many NMPs were detected in DEN-induced hepatocellular carcinomas. These results suggest that the alteration of the NMPs may be involved in DEN-induced hepatocarcinogenesis. PMID- 10378788 TI - Response of DNA ploidy to chemotherapy in primary and metastatic lesions in human osteosarcomas. AB - Primary and pulmonary metastatic and pulmonary metastatic tumors (two synchronous and seven metachronous metastases) in nine patients with osteosarcomas were studied by DNA cytofluorometry. All patients were treated with both pre and postoperative chemotherapy. The results showed that all five diploid osteosarcomas and three of the four aneuploid tumors did not markedly change their ploidy pattern after preoperative chemotherapy, and had almost the same ploidy patterns as the pulmonary metastatic lesions. Those eight tumors showed poor histologic response and chemoresistance by the doxorubicin binding assay. Only one aneuploid osteosarcoma showing good histologic response and chemosensitivity changed its ploidy pattern to diploid, with the disappearance of aneuploid tumor cells and its synchronous pulmonary metastatic tumor also showed conversion to a diploid pattern with massive tumor necrosis. It is evident that those tumors showing no change in their ploidy pattern after chemotherapy were resistant to the chemotherapy. Therefore, we conclude that regardless of whether the pulmonary metastatic tumors were synchronous or metachronous, they showed the same change in their ploidy pattern as well as their chemosensitivity as the primary human osteosarcoma from which they were derived. PMID- 10378789 TI - Differences in metallothionein expression in transplantable mouse mammary tumor lines. AB - In order to elucidate a possible role of metallothionein (MT) in mammary carcinogenesis, MT and sex hormone receptor (estrogen receptor, ER; progesterone receptor, PR) expressions were investigated immunohistochemically in a transplantable pregnancy-dependent mouse mammary tumor (TPDMT-4) and related autonomous tumor sublines (T4-OI96, T4-OI165 and T4-OI320CY) recovered from pregnant and virgin DDD mice. TPDMT-4 showed MT expression in tumor cells, while the expression was less evident in T4-OI165 and T4-OI96 among the autonomous tumor lines; in T4-OI320CY, the MT expression was similar to that in TPDMT-4. Chromatographic study of MT contents in the tumor lines confirmed the results of the immunohistochemical examination. PR and ER were localized in the tumor cells of TPDMT-4, but not in those of autonomous tumor sublines. In TPDMT-4, a significant correlation was observed between MT and ER expressions (r = 0.83, P < 0.01), but not between MT and PR expressions (r = 0.26, P > 0.4), also between MT expression and mitotic activity (r = -0.34, P > 0.3). Since T4-OI96 and T4-OI165 are known to be more malignant than T4-OI320CY, the present study indicates a negative correlation between the MT positivity and progression of the transplantable mammary tumor in mice. PMID- 10378790 TI - Prognostic significance of tumor angiogenesis in epithelial ovarian cancer. AB - Since angiogenesis is considered essential for tumor growth and the development of metastasis, we assessed the correlation of microvessel density (MVD) with overall survival in patients with epithelial ovarian cancer. Histologic slides were immunostained for CD34-antigen. MVD was determined within each tumor by enumeration under a light microscope at 200x magnification and an examination area of 0.25 mm2. The Cox proportional-hazards model was used for multivariate analysis. In 63 patients with epithelial ovarian cancer the 5-year survival rate (OS-%) was as follows: 55.0% (+/-12.5) in 18 patients whose tumors had an MVD < 10/field, and 23.6% (+/-6.7) in 45 patients whose tumors had an MVD(10/field (log rank P = 0.038). MVD showed a significant influence on survival in univariate analysis, but failed to attain a significant value after adjustment for established prognostic parameters such as patients' age at diagnosis, stage of disease, and histologic grading. High MVD was significantly associated with advanced patients' age at diagnosis. This and a considerable heterogenity in the vascular architecture of ovarian carcinoma tissue might be the reasons why MVD did not reveal prognostic significance in multivariate analysis. In contrast to a variety of solid neoplasms, MVD does not seem to be a useful predictor of survival in patients with epithelial ovarian cancer. PMID- 10378791 TI - Analysis of genomic instability in squamous cell carcinoma of the head and neck using the random amplified polymorphic DNA method. AB - Using the random amplified polymorphic DNA (RAPD) method, we identified genomic instability in head and neck squamous cell carcinoma (HNSCC) tissues. We extracted DNA from tumor and corresponding normal tissues of 30 HNSCC patients and amplified with ten random 10-mer arbitrary primers by the RAPD method. Genomic instabilities, which appeared as banding pattern changes between normal and tumor DNA, were detected by at least one primer in all tumor tissues. Moreover, there was significant correlation between the frequency of genomic instability and the degree of tumor differentiation. These results indicate a possible association of genomic instability with malignant potential of head and neck cancer. PMID- 10378792 TI - Wild-type p53 protein potentiates phototoxicity of 2-BA-2-DMHA in HT29 cells expressing endogenous mutant p53. AB - To better understand the effects of p53 on the process of photodynamic therapy (PDT)-induced cell death, we introduced a wild-type p53 gene into the HT29 colorectal carcinoma cell line, which bears an endogenous mutant p53, using a lipofectin system. The influence of p53 status on the sensitivity induced by 2 butylamino-2-demethoxy-hypocrellin A (2-BA-2-DMHA) photosensitization was then examined. The results indicate that infection with wild-type p53 induces a growth arrest but does not induce cell death, and sensitizes the cells to PDT. At a concentration of 5 microM 2-BA-2-DMHA with a red light of 18 J/cm2 (lambda = 600 700 nm), the survival is reduced from 58.72% in HT29 cells to 13.49% in wild-type p53-infected HT29 cells. Apoptosis following PDT appears earlier in HT29 cells infected with wild-type p53 than in parent HT29 cells and empty vector-infected HT29 cells. These findings suggest that although wild-type p53 is, by itself, insufficient to induce apoptosis in cells with p53 mutation, it enhances the photosensitivity of 2-BA-2-DMHA by strongly potentiating the induction of apoptosis. PMID- 10378793 TI - Photochemotherapy of tumours with mesotetrahydroxyphenyl chlorin is pH dependent. AB - Human colon carcinoma cells of the WiDr line were incubated with the tumour localizing and photosensitizing drug mesotetrahydroxyphenyl chlorin (mTHPC) at either pH 6.8, 7.2 or 7.8. The cellular uptake of the drug was found to be independent of the pH value. However, under conditions where the cells contained the same amount of the drug (i.e. after incubation at different pH values) and were exposed to light, their photosensitivity increased with decreasing pH value. Furthermore, the cells were more photosensitive at 4 degrees C than at 37 degrees C. The shoulder on the survive curves, observed for irradiation at 37 degrees C and at pH 7.2-7.8 was practically absent for irradiation at 4 degrees C and even at 37 degrees C when the light exposure took place at pH 6.8. The observations may be related to a reduced repair of photochemotherapy (PCT)-induced damage at 4 degrees C and at pH 6.8. The high PCT efficiency at pH 6.8 may contribute to the tumour selectivity of PCT with mTHPC. PMID- 10378794 TI - Relationship between P-glycoprotein positivity, doxorubicin binding ability and histologic response to chemotherapy in osteosarcomas. AB - We previously reported that the doxorubicin binding ability detected by the doxorubicin (adriamycin) binding assay was closely correlated with the chemosensitivity of human osteosarcomas. In this study, we undertook to clarify the relationship between P-glycoprotein positivity (%PPG) and doxorubicin binding ability (%DB) in human osteosarcomas in order to determine which is a more sensitive index of histologic response to chemotherapy. Ten primary osteosarcomas were analyzed by the doxorubicin binding assay and by immunofluorescence to detect cellular P-glycoprotein positivity. Three good responders to chemotherapy containing doxorubicin showed a %DB greater than 90% (average: 96.43%), whereas the seven poor responders had values less than 80% (average: 35.31%). The difference between the two groups was statistically significant (P = 0.0167). However, the average %PPG of the three good responders was 6.73%, whereas the %PPG of the seven poor responders was 14.27%. There was no significant difference in %PPG between the two groups (P = 0.3051). No negative correlation between the %DB and the %PPG of all osteosarcomas (r = 0.536, P = 0.1104) was found, although there was a trend that those tumors with a high %PPG showed a low %DB. These results suggest that osteosarcomas showing a low %DB and %PPG with poor response to chemotherapy, may have multidrug resistance mechanisms other than P glycoprotein. Therefore, we conclude that doxorubicin binding ability, which reflects all of the doxorubicin-resistant mechanisms, was more sensitive than P glycoprotein positivity in predicting the chemosensitivity of human osteosarcoma. PMID- 10378795 TI - Abnormal frequencies of alleles in polymorphic markers of the 17q21 region is associated with breast cancer. AB - This study was carried out to evaluate the frequencies of alleles of four polymorphic markers in the 17q21 region in breast cancer patients, and their relation to seven pathological parameters. One hundred and sixty-four patients with breast cancer and 102 controls were analyzed. D17S856, D17S855, D17S1323 and D17S1327 polymorphic markers were studied, and used to investigate loss of heterozygosity in this region. The frequencies of alleles at marker D17S856 differed significantly in breast cancer patients and controls, and were related to histologic features considered to indicate a poor prognosis. When present, the pathophenotype of tumors associated with LOH in the 17q21 region is modified. PMID- 10378796 TI - Effect of cisplatin exposure on platinum accumulation and growth inhibition in human neoplastic and normal squamous epithelial cells of the mucosa of the upper aerodigestive tract. AB - The aim of the present study was to investigate how normal head and neck epithelial cells (NHNEC) respond to cisplatin compared to their neoplastic counterparts with respect to intracellular platinum (Pt) levels and growth inhibition. A colorimetric assay was used to assess growth inhibition after exposure to cisplatin for 72 h. Growth inhibition did not differ between cultures of neoplastic (n = 5) and normal cells (n = 5). Intracellular Pt levels, determined with atomic absorption spectroscopy were about 30-fold higher in the normal epithelial cells. The main finding of this study is that normal epithelial cells from the head and neck region have a much higher tolerance for cisplatin than their neoplastic counterparts. Interestingly, this characteristic is without consequence for growth inhibition. PMID- 10378797 TI - CDK-inhibitor olomoucine inhibits cell death after exposure of cell lines to cytosine-arabinoside. AB - Signal transduction for apoptosis or programmed cell death, after DNA damage in mammalian cells, is believed to involve activation of cyclin-dependent kinases (CDKs), especially CDK-1 (cdc2) and CDK-2. We used CDK-inhibitor olomoucine, a purine analogue to evaluate the role CDK inhibition on cytosine-arabinoside (Ara C)-induced cell death. The two drugs showed an antagonistic effect, suggesting that apoptosis after exposure to Ara-C is inhibited by olomoucine. DNA electrophoresis showed a clear inhibition of the apoptotic pattern when olomoucine was added to Ara-C. We conclude that CDK-inhibitor olomoucine inhibits cell death induced by Ara-C. PMID- 10378798 TI - Deletion mapping on the short arm of chromosome 8 in hepatocellular carcinoma. AB - To define the commonly deleted region on chromosome 8p for further positional cloning of the putative tumor suppressor gene, we carried out allelic imbalance (AI) studies in 41 HCCs using a panel of 37 microsatellite markers. The overall AI on 8p was 87.8% (36 of 41). Among the 36 cases with AI, 13 cases showed AI in all of the loci, suggesting entire deletion on the short arm of chromosome 8, while the remaining 23 cases showed partial AI. Detailed deletion mapping identified two independent commonly deleted regions on chromosome 8p. These were as follows: (1) centered by the D8S1819 and D8S1706 loci between the D8S561 and D8S1825 loci, (2) centered by the D8S1733 locus between the D8S298 and D8S1739 loci. These results suggest that the two putative tumor suppressor genes may be present on chromosome 8p. PMID- 10378799 TI - Fabrication, characterization and evaluation of bioceramic hollow microspheres used as microcarriers for 3-D bone tissue formation in rotating bioreactors. AB - Novel bioactive ceramic hollow microspheres with an apparent density in the range 0.8-1.0 g cm(-3) have been developed as microcarriers for 3-D bone tissue formation in rotating-wall vessels (RWV). Hollow ceramic microspheres with a composition of 58-72% SiO2, 28-42% Al2O3 (wt%) and an apparent density 0.8-1.0 g cm(-3) were pretreated in 1.0 N NaOH for 2 h before being coated with synthesized calcium hydroxyapatite (HA) particulate sol. The HA-coated hollow microspheres were sintered for 1 h at 600, 800 and 1000 degrees C. SEM analysis revealed that the grain size and pore size of the calcium phosphate coating increased with the sintering temperature. FTIR analysis showed that crystalline calcium hydroxyapatite was present in the coatings sintered at 600 and 800 degrees C. When sintered at 1000 degrees C, the coating consisted of alpha-tricalcium phosphate. All the coatings adhered well, independent of sintering temperature. The trajectory analysis revealed that the hollow microsphere remained suspended in a rotating-wall vessel (RWV), and experienced a low shear stress (approximately 0.6 dyn cm(-2)). Cell culture studies using rat bone marrow stromal cells and osteosarcoma cells (ROS 17/2.8) showed that the cells attached to and formed 3-D aggregates with the hollow microspheres in a RWV. Extracellular matrix was observed in the aggregates. These data suggest that these hollow bioactive ceramic microspheres can be used as microcarriers for 3-D bone tissue formation in vitro, as well as for the study of the effects of microgravity on bone cell functions. PMID- 10378801 TI - Interface between bone and nacre implants in sheep. AB - We have investigated the interface between bone and chronic implants of nacre in sheep. There was no foreign body reaction over the period of 10 months and the implants were not broken down. Light microscopy indicated activity within an osteoprogenitor cellular layer lining the implant, resulting in a complete sequence of new bone formation. Nacre appeared to bind directly to newly formed bone without any intervening fibrous tissue. Scanning electron microscopy and energy dispersive photon X-microanalysis showed calcium and phosphate ions lining the nacre within the osteoprogenitor tissue. These studies show a dynamic activity of the bone/nacre interface, leading to continuity between the nacre and the bone. PMID- 10378800 TI - Nitrous acid pretreatment of tendon xenografts cross-linked with glutaraldehyde and sterilized with gamma irradiation. AB - Collagenous xenografts made from kangaroo tail tendon cross-linked with glutaraldehyde have a potential application in the reconstruction of massive digital tendon deficits. However, a limitation to the clinical use of these xenografts has been the optimization of collagen cross-linking, and subsequent bio-incorporation and retention of mechanical properties following implantation. The purpose of this study was to evaluate the effect of nitrous acid on modulating the biologic and mechanical properties of tendon xenografts cross linked with glutaraldehyde. Tendon xenografts were pretreated with 0.1 or 0.01 M nitrous acid solution, prior to cross-linking in 2% glutaraldehyde and sterilization by gamma irradiation. Xenografts were implanted intramuscularly in rabbits to examine biocompatability, and also used to repair ovine digital extensor tendon deficits to evaluate functional incorporation. Histologically, intramuscularly implanted nitrous acid pretreated xenografts in rabbits had a greater degree of diffuse cellular infiltration into interstitial splits in the graft than controls after 12 weeks. Xenografts implanted in an ovine extensor tendon deficit were evaluated after 26 and 52 weeks. Rate of failure of tenorrhaphies between host tendon and xenografts overall (15/21) was significantly greater (P < 0.05) than for autografts (1/21), suggesting that the holding power of sutures in xenografts was inferior to that obtained in autografts. Tensile failure stress of midsections of both nitrous acid pretreated and control xenografts was about 100 MPa prior to implantation (time zero). After 26 and 52 weeks, failure stress of both types of xenografts was significantly less than at time zero (P < 0.05). At 52 weeks, failure stress of nitrous acid pretreated xenografts (47.4 +/- 3.1 MPa) was significantly less than control xenografts (63.7 +/- 5.4 MPa); (P < 0.05). However, nitrous acid pretreated xenografts were similar to control xenografts in failure load (357 +/- 29 and 354 +/- 26 N, respectively), but they tended to have larger cross-sectional areas (7.6 +/- 0.5 versus 5.7 +/- 0.6 mm2, respectively) which were responsible for the lower calculated value for failure stress. Histologically, autografts maintained their normal tissue architecture and evoked a more limited cellular response in surrounding tissues than xenografts (P < 0.05). Both types of xenograft were surrounded by a thicker cuff of cellular response than autografts. However, compared to control xenografts, nitrous acid pretreated xenografts had more extensive fragmentation and splitting of collagen bundles, and more diffuse cellular and vascular infiltration into these interstitial splits, and these alterations were apparently contributing to the greater 'swelling' of these xenografts. It was concluded that pretreatment of tendon xenografts with nitrous acid modulated their biologic and material properties. Further studies are needed to elucidate the mechanism of these effects, and to determine if the protocol for tendon xenograft preparation could be optimized for improved clinical performance. PMID- 10378802 TI - Lipid uptake in synthetic vascular prostheses explanted from humans. AB - Previous in vivo studies in humans and dogs have revealed an atherosclerosis-like phenomenon in which lipid penetration within arterial prosthesis wall was observed. The primary goal of the present study was therefore to investigate the occurrence of this lipid retention in ePTFE prostheses implanted in humans and therefore identify potential risk factors related to this phenomenon. Lipid uptake in 367 ePTFE microporous vascular prostheses explanted from humans was studied using Fourier transform infrared spectroscopy. The assignment of the infrared absorption features clearly revealed the presence of strongly bonded unsaturated fatty acids to the microporous structure of the prostheses. A one-way ANOVA statistical analysis showed that the lipid uptake in the synthetic vascular prostheses depended on the duration of implantation of the prosthesis and on the sex of the patient. A two-way ANOVA showed that a relationship existed between the estimated lipid uptake and the internal diameter of the prosthesis. These results confirm that the lipid uptake phenomenon depends on some clinical factors related either to the patients or to the prostheses' morphological parameters. PMID- 10378803 TI - Amphiphilic diblock copolymeric nanospheres composed of methoxy poly(ethylene glycol) and glycolide: properties, cytotoxicity and drug release behaviour. AB - Amphiphilic diblock copolymers based on methoxy poly(ethylene glycol) (MePEG) and glycolide with different molar composition were synthesized by bulk polymerization without any catalysts. Using diblock copolymers, we have prepared indomethacin-loaded polymeric nanospheres by forming a micelle in selective solvents. The size of nanospheres measured using dynamic light scattering exhibited a narrow monodisperse size distribution and an average diameter in the range of less than 200 nm. The critical micelle concentration (CMC) of MG70 sample determined by fluorescence spectroscopy was 1.57 x 10(-7) mol/l which was lower than the CMC of common low molecular weight surfactant. In vitro release experiments using indomethacin-loaded MePEG/glycolide nanospheres exhibited the sustained release behaviour without any burst effect. In addition, the results of cytotoxicity tests using an MTT assay method showed that these MePEG/glycolide nanospheres could remarkably reduce cell damage compared with unloaded free drug. PMID- 10378804 TI - Extracellular matrix protein-induced changes in human salivary epithelial cell organization and proliferation on a model biological substratum. AB - We have used a denuded rat tracheal preparation as a biological substratum on which to examine the growth and morphology of a salivary epithelial cell line (HSG) in vitro. In the absence of an additional coating of matrix proteins, HSG cells grew at low density on tracheae. Coating the tracheae with Vitrogen (a commercial collagen I preparation) or fibronectin promoted HSG cell growth and monolayer formation. Conversely, if a coating of Matrigel was applied, cells grew in a more organized fashion, but at low density. Generally similar results were obtained with cells grown on laminin and collagen IV but with less organization. These studies demonstrate the utility of a natural, tubular substratum for testing the influence of different matrix proteins on salivary epithelial cell behavior. PMID- 10378805 TI - Nanoindentation studies of titanium single crystals. AB - Titanium single crystal planes of different atomic density have been reported to show different oxidation characteristics. The differences in oxide characteristics have further been demonstrated to lead to differences in osteoblast attachment. Investigations of the preferred crystallographic planes of titanium for osteoblast attachment can be used to optimize the surfaces of single crystal and polycrystalline titanium implants for anchoring various prostheses. Nanoindentation techniques were used to determine mechanical properties of two crystallographic planes of titanium of different atomic density. Modulus of elasticity of 128 +/- 10 GPa was obtained for polycrystalline titanium and 123 +/ 5 and 124 +/- 6 GPa for the basal plane and pyramidal planes, respectively. The variation of modulus with crystal orientation was not greater than the statistical variation in the data. Surface hardness values were 2.1 +/- 0.1 GPa for the polycrystalline sample and 1.6 +/- 0.1 and 1.9 +/- 0.1 GPa, respectively, for the basal and pyramidal planes. Curves of hardness as a function of depth (0 2000 nm) obtained from electrochemically polished surfaces showed a sharp increase at shallow depths and may reflect changes caused by oxidation of the titanium surfaces. PMID- 10378806 TI - Factors affecting the degradation rate of poly(lactide-co-glycolide) microspheres in vivo and in vitro. AB - The purpose of this work was to study the degradation of poly(lactide-co glycolide) (PLG) microspheres in vivo and in vitro. Degradation rate constants were determined by measuring the polymer molecular weight as a function of time by gel-permeation chromatography. The effects of PLG chemistry and the effects of encapsulating the sparingly soluble salt zinc carbonate and the protein recombinant human growth hormone (rhGH) on the degradation rate were assessed. It was found that in vivo degradation was faster than in vitro degradation. In addition, different types of PLGs were found to degrade at different rates depending on the chemistry of the polymer end group and, to a lesser extent, the molecular weight. Finally, zinc carbonate was found to retard the degradation of some PLGs. These degradation studies have proved valuable in the design of sustained release microsphere products. PMID- 10378807 TI - A formal risk assessment of silicone breast implants. AB - In 1992, the United States Food and Drug Administration (FDA) announced that breast implants filled with silicone gel would be available only through controlled clinical studies despite the fact that they had been used for mammoplasty in millions of women around the world for more than 30 years. The safety of silicone gel breast implants had come into question after several reports on a possible association between the implants and subsequent development of connective-tissue diseases. Risk assessment refers to the systematic, scientific characterization of potential adverse effects of human exposures to hazardous agents or activities. The following risk assessment is intended to review the current scientific evidence for the safety of silicone gel-filled breast implants since the FDA's decision in 1992. There now appears ample evidence from the scientific literature for the safety of these prostheses. PMID- 10378808 TI - The treatment of type 2 diabetes: good news from the UK. PMID- 10378809 TI - Acalculous gallbladder pain: a largely unrecognised entity. AB - AIM: To study the presentation for and outcome of cholecystectomy in patients with acalculous gallbladder pain. METHODS: Sixty-six consecutive patients with prospective documentation underwent cholecystectomy for putative acalculous gallbladder pain between December 1988 to April 1995. The diagnosis was made on clinical grounds, but in the majority, a CCK oral cholecystogram was performed. Outcomes were assessed by postal questionnaire mailed in October 1995 or by the last recorded follow- up. RESULTS: Fifty-eight females and eight males, with a median age of 37.5 years had experienced abdominal pain, usually with associated nausea, for a median of three years. Preoperative investigations were non contributory, with the exception of the CCK oral cholecystogram which was regarded as abnormal in all instances. At a median follow-up of 40 months, 48 patients (72.7%) described their symptoms as either totally relieved or much improved by cholecystectomy. CONCLUSION: Though the pathophysiology remains poorly understood, there is clearly a group of patients who suffer from gallbladder pain in the absence of gallstones and who benefit from cholecystectomy. PMID- 10378810 TI - Dealing with a bleeding nuisance: a study of haemophilia care in New Zealand. AB - AIM: To describe the social and demographic characteristics of people with haemophilia in New Zealand and to identify key issues in their treatment. METHODS: The research (1994-6) used a combination of quantitative and qualitative methods, including key person interviews and focus groups, a national survey (n=193), interviews (80), participant observation and consultative report writing (30 participant-consultants). RESULTS: The study population was mainly (95%) male, somewhat younger than the general New Zealand population, and was 90% Caucasian but otherwise conformed closely to the socio-demographic profile of the general population. Participants clustered towards the severe end of the condition of haemophilia. There was great variation between participants in incidence of bleeds. Legs and arms were most commonly affected. The majority (61%) of bleeds were treated within six hours of detection. Despite considerable advances and an emphasis on the provision of adequate and prompt treatment, there is still much room for improvement to obviate problems such as joint damage, pain and the need for surgery. CONCLUSIONS: People with haemophilia and haemophilia health service providers struggle to achieve optimal care. PMID- 10378811 TI - Health advice given by general practitioners for travellers from New Zealand. AB - AIMS: To investigate where general practitioners (GP's) in New Zealand view travel health advice best given and where they refer for this advice, the prevalence of travel health advice reported to be given, and the prevalence of written advice, including a doctor's letter. METHOD: This was a descriptive cross sectional study, using self-report questionnaires, sent to 400 GPs randomly selected from the register of the New Zealand Medical Council. RESULTS: Three hundred and thirty-two GPs (83%) responded. Most GPs reported that they saw travel medicine as best practised in general practice (241/308, 78%) or in a combination of locations, usually including general practice (28/308, 9%). Most GPs (223/308, 72%) did not refer travellers for travel health advice. Health advice concerning malaria (310/310, 100%), immunisation (309/310, 100%), travellers' diarrhoea (296/305, 97%), insect avoidance (287/ 299, 96%), sexually transmitted diseases/human immunodeficiency virus (233/283, 82%), water purification (235/293, 80%) and other areas (35/75, 47%) was given. Written advice was usually given by 23% of GPs (69/302). Written advice was significantly more likely to be provided by those GPs with an interest in travel medicine (chi2=5.67, df=1, p<0.005), experience in tropical medicine/developing countries (chi2=6.69, df=1, p<0.001), a policy on travel medicine (chi2=21.4, df=1, p<0.001), a written policy on travel medicine (chi2=302.0, df=1, p<0.001), who saw a higher number of travellers per week (t=-2.51, df=296, p<0.05) and who saw a significantly higher proportion of patients who were travellers (t=-3.27, df= 295, p=0.001). Almost all GPs (303/310, 98%) reported giving their travelling patients a doctor's letter at least sometimes but only 7% (23/310) always gave travellers a doctor's letter. GPs with training in travel medicine/related area were significantly more likely to provide travellers with a doctor's letter (chi=11.61, df=3, p<0.01). CONCLUSIONS: This study confirmed that GPs in New Zealand see travel health advice as best given in general practice. Travel health advice, as recommended by New Zealand guidelines, should continue to be given. With limited time in general practice to advise travellers, GPs should also consider giving written advice, including a doctor's letter, more often. Epidemiological and specialist support by public health units and commercial groups, continuing medical education and training in travel medicine for GPs are among the major considerations. Further studies are needed concerning the adequacy and currency of destination-specific advice for travellers. PMID- 10378812 TI - An evaluation of pharmaceutical management and budget holding in Pegasus Medical Group. AB - AIMS: To describe and evaluate pharmaceutical management, including budget holding, in Pegasus Medical Group (Pegasus), to determine savings being achieved, to analyse variation in prescribing behaviour and to compare the findings with national and international experience. METHODS: Trends in pharmaceutical expenditure of the 150 Pegasus' 208 members who had a continuous prescribing record for the three years ending December 1996 were compared with national trends. Expenditure per member, per consultation and per item were also analysed. RESULTS: Pegasus has implemented a comprehensive and classical pharmaceutical management strategy. This includes active personalised feedback, information sharing, peer review groups and information system development, all within an incentive framework of retained savings for new services. Although about 5% savings of total pharmaceutical expenditure were identified by the above method, the real level may be higher. Wide variation between members in their prescribing behaviour was explained almost entirely by the volume rather than the price of the drugs prescribed. Targeting of the volume issue is therefore likely to have a much more significant effect in reducing inappropriate variation. CONCLUSION: The results indicate that the achievements of Pegasus, as for other independent practitioner associations, go far beyond the modest level of pharmaceutical savings achieved. These include the development of a substantial infrastructure, peer review processes, new internal and external relationships and accountability for the management of both quality and cost in what may be styled clinical governance. Such achievements put Pegasus and other independent practitioner associations into a strong position to take on new initiatives including integration with secondary care. PMID- 10378813 TI - Antimicrobial resistance. PMID- 10378814 TI - Health and Disability Commissioner Act 1994. PMID- 10378815 TI - Assessment for ACC cover. PMID- 10378816 TI - Should folate be added to flour. PMID- 10378817 TI - Youth suicide report 1997. PMID- 10378818 TI - Melioidosis--an emerging disease in New Zealand? PMID- 10378819 TI - Intravenous hydroxyethylrutosides combined with long-term oral anticoagulation in atherosclerotic nonreconstructable critical leg ischemia: a retrospective study. AB - OBJECTIVE: To evaluate in a group of seriously diseased patients with nonreconstructable chronic critical leg ischemia (CLI), treated by a combination of i.v. hydroxyethylrutosides (HR)* and oral anticoagulation (AC) by warfarin, the short-term effects on the cutaneous microvascular blood perfusion of the soles of feet and especially the long-term clinical outcome in terms of amputation and death. DESIGN: A retrospective comparison between two groups of patients, HR + AC and a comparable reference group, fulfilling the same inclusion and exclusion criteria corresponding to the definition of CLI according to the Second European Consensus Document (1991). Clinical follow-up in both groups was made after 1, 3, 6, 12, and 24 months. SETTING: Patients were examined at university departments of clinical physiology with special interest in peripheral vascular disease, in cooperation with colleagues at university departments of surgery, internal medicine and dermatology of Karolinska Hospital, Sodersjukhuset and Huddinge Hospital. PATIENTS: A total of seventy patients with CLI according to the definition of the Second European Consensus Document, 1991, ie, besides severe rest pain or ischemic lesions also a toe blood pressure < 30 mg Hg. Group with HR + anticoagulation (AC): 42 patients (19 diabetics, 23 nondiabetics). Reference group: 28 patients (18 diabetics, 10 nondiabetics). For distribution of age and toe blood pressure at baseline, see Table I. INTERVENTIONS: Therapy group: besides ordinary standard therapy, daily HR infusions for a mean period of 3.6 weeks + oral anticoagulation continued to the end of the study at 24 months. A comparable reference group on the same basic therapy but without the combination HR + AC. PARAMETERS IN EVALUATION: Short-term parameters: clinical data, skin temperature, and fluorescein imaging. Long-term outcome: amputation or death. RESULTS: Short-term and long-term results with HR + AC indicated that patients with severe CLI and very poor prognosis benefited in terms of survival and limb salvage from initial therapy with HR infusion combined with long-term oral anticoagulation. Results of this combined treatment seem at least comparable with those with i.v. prostacyclin analogies. PMID- 10378820 TI - Combination of calcium channel blockers and beta blockers for patients with exercise-induced angina pectoris: a double-blind parallel-group comparison of different classes of calcium channel blockers. The Netherlands Working Group on Cardiovascular Research (WCN). AB - The combination of calcium channel blockers and beta blockers is more effective for the treatment of exercise-induced angina pectoris than beta blocker monotherapy. Since ischemia in exercise-induced angina is essentially preceded by an increase in heart rate, calcium channel blockers with negative chronotropic property may perform better for this purpose than nonchronotropic compounds. A 335-patient, 10-week, double-blind, parallel-group comparison of amlodipine 5 and 10 mg, diltiazem XR 200 and 300 mg, and mibefradil 50 and 100 mg treatment added to baseline beta blocker treatment was performed. Exercise testing (ETT) was performed by bicycle ergometry. Although none of the calcium channel blockers improved duration of exercise or amount of workload, all of them significantly delayed onset of 1 mm ST segment depression on ETT (p<0.001 for any treatment versus baseline). In addition, mibefradil, both low- and high-dose treatment, produced the largest delays (low dose: different from diltiazem and amlodipine by 24.1 and 29.8 s, p<0.003 and <0.001, respectively; high dose: different from diltiazem and amlodipine by 33.7 and 37.0 s, p<0.001 and <0.001, respectively). These effects were linearly correlated to the amount of rate pressure product (RPP) reduction. Serious symptoms of dizziness likewise occurred significantly more frequently with mibefradil (p<0.05) and led 19 patients taking mibefradil to withdraw from the trial. The authors conclude that calcium channel blockers with negative chronotropic property provide better delay of ischemia in patients with exercise-induced angina but that the concomitant risk of intolerable dizziness largely reduces this benefit. PMID- 10378821 TI - Effects of triflusal on arteriosclerosis progression assessed with high resolution arterial ultrasound. AB - In order to evaluate the effect of triflusal (2-acetyloxy-4-trifluoromethyl benzoic acid), an orally active antiplatelet agent, on arteriosclerosis progression, a pilot, parallel, double-dummy, double-blind clinical trial vs acetylsalicylic acid (ASA) was carried out in patients with subclinical atherosclerotic lesions. The trial consisted of a 2-week run-in placebo phase, followed by a 12-month oral treatment with triflusal (600 mg/day) or ASA (300 mg/day). The primary variable was identified in the ultrasonic biopsy (UB) score; the secondary variables were the UB class changes of each arterial site, the rate of progression (ROP), the intima-media thickness (IMT), and the symptoms of arteriosclerosis. Data were evaluated by use of analysis of variance and Chi square test. Forty-three patients (31 men, 12 women, mean age 62.8 +/- 8.4 SD) were randomized to triflusal (15 men, 6 women, mean age 64.3 +/- 6.7) or to ASA (16 men, 6 women, mean age 61.3 +/- 9.6). The analysis of variance on the UB score showed no difference between treatments: the patients' UB scores remained unchanged with no progression, thus indicating that no patient worsened during treatment. When all arterial sites under evaluation are considered, 86% of the sites in the triflusal group and 85% in the ASA group remained unchanged. No relevant change was recorded in vital signs and routine laboratory tests. Gastric disturbances were reported by two and three patients treated with triflusal and ASA, respectively. In conclusion, triflusal appears as effective as ASA in slowing arteriosclerosis progression. PMID- 10378822 TI - Concomitant use of intraaortic balloon counterpulsation and streptokinase in acute anterior myocardial infarction. AB - Using a prospective, nonrandomized design, the authors sought to determine whether concomitant use of intraaortic balloon counterpulsation (IABP) and streptokinase in acute anterior myocardial infarction (MI) would improve the in hospital mortality rate and angiographic findings. The study included 45 patients with an acute anterior MI. All patients received intravenous streptokinase. Among these, 25 patients had concomitant IABP while the remaining 20 patients had streptokinase alone. All patients underwent cardiac catheterization. Patients treated with concomitant IABP had a significantly higher frequency of thrombolysis in myocardial infarction (TIMI) grade 3 flow (n: 11; 44% vs n: 1; 5%, p<0.05), and there was a trend toward a lower in-hospital mortality rate in the IABP group (n: 0; 0% vs n: 3; 15%, p=0.08). The angiographic presence of thrombus image and grade > or =2 coronary collateral circulation to the infarct related coronary artery for the IABP and non-IABP groups did not differ significantly. The preliminary results of this study suggest that concomitant use of IABP and streptokinase in acute anterior MI increases the incidence of TIMI grade 3 flow and may have decreased the in-hospital mortality rate without unacceptable rates of vascular or hemorrhagic complications. PMID- 10378823 TI - Peripheral vascular resistance limits exercise functional capacity of mild hypertensives. AB - To evaluate the physiological basis for suboptimal peak exercise oxygen consumption (VO2p) observed in the early stage of hypertension, 25 WHO Stage I hypertensive men with normal left ventricular mass and 10 healthy control subjects of equivalent age underwent the maximal cardiopulmonary exercise test with contemporary measurement of cardiac output with Tc99m angiocardiography. At peak exercise hypertensive patients had lower VO2p (p < 0.045) and cardiac output (p < 0.014) and higher vascular resistance (p < 0.010) than controls. At multiple regression analysis VO2 was positively related to cardiac output in controls (r = 0.80, p < 0.02), whereas in hypertensives the best (negative) correlation was observed with peripheral vascular resistance (r = -0.72, p < 0.04). Thus reduced cardiopulmonary function during physical exercise in hypertensives seems to be mainly related to impaired peripheral vascular autoregulation. PMID- 10378824 TI - Nitric oxide impacts endothelin-1 gene expression in intrapulmonary arteries of chronically hypoxic rats. AB - This study aimed to investigate whether nitric oxide (NO) could inhibit the elevated endothelin-1 (ET-1) gene expression by pulmonary artery endothelial cells or smooth muscle cells in chronically hypoxic rats by use of in situ hybridization. Male Wistar rats (n = 40) were randomly divided into 1-week hypoxia group, 1-week hypoxia with L-arginine (L-arg) group, 1-week hypoxia with N(omega)-nitro-L-arginine methyl ester (L-NAME) group, 2-week hypoxia group, 2 week hypoxia with L-arg group, and 2-week hypoxia with L-NAME group. All rats were put into a normobaric hypoxic chamber with an oxygen concentration of 10 +/- 0.5% for hypoxic challenge. The results showed that most pulmonary arteries had 1 50% of the endothelial cells showing positive signals for ET-1 expression in hypoxic rats, which was significantly suppressed by L-arg. L-NAME, however, significantly augmented ET-1 gene expression in pulmonary artery endothelial cells and smooth muscle cells. The results suggest that endogenous NO markedly inhibits ET-1 mRNA expression in both pulmonary artery endothelial cells and smooth muscle cells in chronically hypoxic rats, which may be one of the mechanisms by which NO modulates hypoxic pulmonary circulation. PMID- 10378825 TI - Basic fibroblast growth factor increases regional myocardial blood flow and salvages myocardium in the infarct border zone in a rabbit model of acute myocardial infarction. AB - Basic fibroblast growth factor (bFGF) has been shown by some to promote angiogenesis and myocardial salvage in experimentally induced acute myocardial infarction. Although these findings have spurred much clinical interest, they are not universally observed, and the true efficacy of bFGF remains unclear. The authors used a rabbit model of acute myocardial infarction to further elucidate the effects of bFGF on acutely infarcted myocardium containing few collaterals. Myocardial infarction was evoked by ligation of the left coronary artery. Prior to ligation, either 100 microg of bFGF (bFGF group; n = 15) or physiological saline (control group; n = 22) was injected into the myocardium supplied by the ligated artery. With use of nonradioactive colored microspheres, regional blood flow (Qm) was measured before, immediately after, and 4 weeks after coronary artery ligation. Infarct and border zone sizes were measured in cross-sectional slices of the resected hearts, and the amount of viable myocardium (myocardium score) and the extent of fibrosis were histologically determined in each area. Four weeks after ligation, Qm values in the infarcted area did not significantly differ between the bFGF and control groups (0.54 +/- 0.36 vs 0.48 +/- 0.30 mL/min/g); in the border zone, Qm tended to be higher in the bFGF group (3.39 +/- 2.68 vs 1.47 +/- 0.80 mL/min/g), but the difference was not significant; finally in the noninfarcted area, Qm was significantly (p < 0.05) higher in the bFGF group (6.06 +/- 3.85 vs 2.09 +/- 0.82 mL/min/g). There was no significant difference in the amount of viable myocardium or the extent of fibrosis in the infarcted areas of the two groups. In the border zone, however, the amount of viable myocardium was significantly (p < 0.005) larger in the bFGF group (61.8 +/ 8.5% vs 35.8 +/- 20.3% of the visual field). Likewise, as graded on a scale from 0 to 5, the extent of fibrosis was significantly (p < 0.005) less in the bFGF group (2.1 +/- 0.5 vs 3.3 +/- 0.8). In conclusion, injection of bFGF into acutely infarcted myocardium increased blood flow to the noninfarcted area and salvaged the myocardium in the border zone. PMID- 10378826 TI - Saphenous vein graft ectasia: an unusual late complication of coronary artery bypass surgery. A case report. AB - Aneurysms and ectasias of saphenous vein grafts are infrequent complications of coronary artery bypass surgery. They usually present as an expanding asymptomatic mediastinal mass on chest x-ray film or computed tomography scan. Though rare, they must be excluded from the differential diagnosis of mediastinal masses to avoid potentially dangerous needle biopsy. The authors describe ectasia of a saphenous vein graft in a 62-year-old man 14 years after coronary artery bypass surgery. The relevant literature is also discussed. PMID- 10378827 TI - Coronary artery aneurysms, aortic dissection, and hypertension secondary to primary aldosteronism: a rare triad. A case report. AB - Primary aldosteronism is a relatively uncommon etiology of hypertension. Plasma renin activity is suppressed in the majority of the cases but not always. Plasma renin activity has been associated with increased vascular injury. The occurrence of vascular complications has rarely been reported with low plasma renin activity. The authors report a case of long-standing secondary hypertension due to primary aldosteronism with coronary artery aneurysms and aortic dissection. Diagnosing is important, for therapeutic intervention can be curative. PMID- 10378828 TI - Multivessel spontaneous coronary artery dissection in a patient with severe systolic hypertension: a possible association. A case report. AB - Spontaneous coronary artery dissection (SCAD) is an uncommon cause of myocardial ischemia and infarction. Hypertension has not been associated with SCAD. The authors report multivessel SCAD in an elderly woman with severe systolic hypertension. They postulate that hypertension of this degree may play a pathophysiologic role in the causation of SCAD. PMID- 10378829 TI - Trousseau's syndrome with brachiocephalic vein thrombosis in a patient with uterine carcinosarcoma. A case report. AB - The authors treated a patient with the previously unreported occurrence of brachiocephalic vein and superior vena cava thrombosis in association with a distantly located cancer. A 71-year-old woman presented with swelling over the right side of the neck and abdominal distension. Physical examination revealed a huge mass, and computed tomography demonstrated thrombosis of the brachiocephalic vein and superior vena cava accompanied by jugular vein dilatation. No coagulation disorder was demonstrable. After anticoagulation and thrombolysis, hysterectomy was performed; microscopic examination of the specimen revealed uterine carcinosarcoma. Even though local tumor obstruction is a much more common cause of neck vein thrombosis, a distant occult cancer can present as this form of Trousseau's syndrome. In patients with otherwise unexplained neck vein thrombosis, examination not only of the head and neck but also of the abdomen and pelvis should be pursued. PMID- 10378830 TI - Successful transcatheter closure of a patent ductus arteriosus: using two Gianturco coils in a 41-year-old woman. A case report. AB - Patent ductus arteriosus (PDA) is a form of congenital heart disease uncommonly diagnosed in adult patients. Transcatheter closure of PDA has been widely used in children. However, the experience is limited in adults especially with use of Gianturco coils. The authors describe a case of successful transcatheter closure of a PDA, incidentally diagnosed in a 41-year-old woman, by successively deploying two coils by a transarterial approach. No residual shunting was seen angiographically after the procedure. A literature review of similar procedures in adult patients is discussed. PMID- 10378831 TI - Blood from patients with hereditary hemochromatosis--a wasted resource. PMID- 10378832 TI - Hemochromatosis and blood donors: a perspective. PMID- 10378833 TI - Allogeneic blood progenitor cell collection in normal donors after mobilization with filgrastim: the M.D. Anderson Cancer Center experience. AB - BACKGROUND: Information on the safety and efficacy of allogeneic peripheral blood progenitor cell (PBPC) collection in filgrastim-mobilized normal donors is still limited. STUDY DESIGN AND METHODS: The PBPC donor database from a 42-month period (12/94-5/98) was reviewed for apheresis and clinical data related to PBPC donation. Normal PBPC donors received filgrastim (6 microg/kg subcutaneously every 12 hours) for 3 to 4 days and subsequently underwent daily leukapheresis. The target collection was > or =4 x 10(6)CD34+ cells per kg of recipient's body weight. RESULTS: A total of 350 donors were found to be evaluable. Their median age was 41 years (range, 4-79). Their median preapheresis white cell count was 42.8 x 10(9) per L (range, 18.3-91.6). Of these donors, 17 (5%) had inadequate peripheral venous access. Leukapheresis could not be completed because of apheresis-related adverse events in 2 donors (0.5%). Of the 324 donors evaluable for apheresis yield data, 221 (68%) reached the collection target with one leukapheresis. The median CD34+ cell dose collected (first leukapheresis) was 462 x 10(6) (range, 29-1463). The main adverse events related to filgrastim administration in donors evaluable for toxicity (n = 341) were bone pain (84%), headache (54%), fatigue (31%), and nausea (13%). These events were rated as moderate to severe (grade 2-3) by 171 (50%) of the donors. In 2 donors (0.5%), they prompted the discontinuation of filgrastim administration. CONCLUSION: PBPC apheresis for allogeneic transplantation is safe and well tolerated. It allows the collection of an "acceptable" PBPC dose in most normal donors with one leukapheresis, with minimal need for invasive procedures. PMID- 10378835 TI - A new apheresis procedure for the preparation of high-quality red cells and plasma. AB - BACKGROUND: Multicomponent apheresis is an alternative way of preparing blood components that avoids the delay between collection and separation seen with standard whole-blood techniques. STUDY DESIGN AND METHODS: An apheresis device has been modified to facilitate the combined collection of a unit (250 mL) of red cells (RBCs) and a high-volume unit (475 mL) of plasma. The procedure, using 8 percent ACD-A, has been tested in two European blood centers. Each center performed 20 procedures for in vitro evaluation of collected RBCs and plasma and 10 procedures for evaluation of in vivo RBC recovery. All RBCs were white cell reduced by filtration. One-half of the RBC units were stored in the additive solution Adsol and one-half in another such solution (Erythro-Sol). RESULTS: The target volumes of RBCs and plasma were obtained in 27 minutes (range, 20-44 min) by using three to six cycles in a single-needle procedure. Saline (275 mL) was used to replace fluid volume withdrawn in excess of standard whole-blood donation. No side effects occurred, with the exception of minor signs of hypocalcemia. RBC ATP was well maintained (>65% at Day 42) during storage; 2,3 DPG was less well maintained, with virtually none remaining at Day 21 in either Adsol or Erythro-Sol. The RBC in vivo recoveries, after 42 days of storage at 4+/ 2 degrees C determined by the single-label method, were 86.7+/-7.2 percent (Erythro-Sol) and 84.4+/-8.1 percent (Adsol). Mean plasma factor VIII levels were >100 percent in all test groups. CONCLUSION: A novel automated technique for the simultaneous collection and preparation of RBCs and plasma has been evaluated. The apheresis procedure was acceptable and well tolerated by donors, and it resulted in high-quality blood components. Further optimization of the system should yield a practicable component suitable for routine use in blood banks. PMID- 10378834 TI - The timing of granulocyte-colony-stimulating factor administration after chemotherapy does not affect stem and progenitor cell apheresis yield: a retrospective study of 65 cases. AB - BACKGROUND: The optimal time for postchemotherapy granulocyte-colony stimulating factor (G-CSF) administration before peripheral blood stem and progenitor cell (PBPC) collection is not well defined. The impact of G-CSF scheduling on the number of CD34+ cells collected by leukapheresis from 65 patients with malignant disease was studied retrospectively. STUDY DESIGN AND METHODS: Chemotherapy was performed on Days 1 and 2 and was followed by G-CSF to mobilize PBPCs. In Group 1, 30 patients received the first dose of G-CSF immediately after the end of chemotherapy, as commonly recommended. In Group 2, 35 patients received the first G-CSF dose after the end of chemotherapy (Days 7 or 8). RESULTS: No difference was observed between the two groups in white cell recovery and the median number of CD34+ cells harvested. The number of leukapheresis procedures necessary to obtain the minimal number of 3 x 10(6) CD34+ cells per kg was the same. The proportion of patients with a failure of PBPC collection was similar, and G-CSF consumption was reduced in Group 2 without increasing infectious risks. CONCLUSION: Early administration of G-CSF after chemotherapy appears not to be a prerequisite for satisfactory PBPC collection. This approach could allow significant savings in terms of medical cost. A randomized and prospective study would be necessary, however, to assess the validity of these conclusions. PMID- 10378836 TI - White cell reduction during plateletpheresis: a comparison of three blood cell separators. AB - BACKGROUND: White cell (WBC)-reduced single-donor platelet concentrates (SDPs) can be collected by the newest generation of blood cell separators. Three WBC reduction techniques during plateletpheresis were investigated in the present study with respect to WBC content and platelet yield. STUDY DESIGN AND METHODS: The Amicus device used the elutriation principle for WBC reduction, and separations with periodically alternating interface position (PAIP) were employed in the AS.TEC 204. WBC reduction by in-line filtration was performed in the MCS+. Platelets were measured electronically and WBCs were determined manually (Nageotte chamber). RESULTS: In-line filtered SDPs showed significantly lower WBC content (0.088+/-0.178 x 10(6)) than SDPs that were WBC reduced by elutriation (0.31+/-0.48 x 106) or PAIP technique (0.89+/-1.57 x 10(6), p = 0.0001). Platelet yield (5.0+/-0.46 x 10(11)) was significantly higher in components obtained with the Amicus device (p = 0.0001). The AS.TEC 204 and MCS+ gave similar results for platelet yields: 3.15+/-0.63 and 3.28+/-0.71 x 10(11), respectively. CONCLUSIONS: The plateletpheresis systems studied allow the collection of WBC-reduced SDPs. In line filtration resulted in the best WBC reduction. Some SDPs collected with the devices studied had a WBC content >1 x 10(6) per unit. Platelet yield was significantly higher in SDPs from the Amicus device. PMID- 10378837 TI - Hemochromatosis probands as blood donors. AB - BACKGROUND: There has been no estimate of the potential eligibility of hemochromatosis probands or patients as blood donors or the suitability for transfusion of their blood that was removed by therapeutic phlebotomy. STUDY DESIGN AND METHODS: According to guidelines of the American Association of Blood Banks, a retrospective estimate of these factors in 211 adult white hemochromatosis probands diagnosed during routine medical care was performed. The findings were compared to those in volunteer white whole-blood donors. RESULTS: Before diagnosis of hemochromatosis, 49 probands had voluntarily donated 597 units of blood; 88 percent were donated by men. After diagnosis, 142 (67%) of 211 probands were potentially eligible. Data on each unit removed during iron depletion therapy and during the first year of maintenance therapy (therapeutic phlebotomy) were available in 86 eligible probands. Of 1592 units, 1029 (65%) obtained during iron-depletion therapy in eligible probands were potentially suitable; 86 percent were from men. During maintenance therapy, 106 (88%) of 121 units from eligible probands were potentially suitable. In volunteer donors, 255,567 (94%) of 273,302 presenting donors were accepted. After testing and laboratory losses, 239,300 (94%) units were acceptable for transfusion. CONCLUSIONS: In comparison with normal volunteers, hemochromatosis probands at diagnosis are less likely to be eligible as blood donors. The percentage of units obtained from patients during iron-depletion therapy that are suitable for transfusion is also lower, although the percentage increases during maintenance therapy. PMID- 10378838 TI - Corrected count increment and percent platelet recovery as measures of posttransfusion platelet response: problems and a solution. AB - BACKGROUND: Corrected count increment (CCI) and percent platelet recovery (PPR) are measures of response to platelet transfusion that "correct" the count increment for blood volume and number of platelets transfused. Their potential for data distortion is described, and a regression analysis is suggested that is more informative and avoids the inherent problems associated with using ratios as outcome measures. STUDY DESIGN AND METHODS: Data from the first platelet transfusion for 585 patients from the Trial to Reduce Alloimmunization to Platelets (TRAP) were used to model methods of analyzing posttransfusion platelet response. RESULTS: By linear regression analysis, unfiltered platelet components gave a greater posttransfusion increment on average (p = 0.001), but filtered platelets gave a greater increment per platelet transfused (p = 0.003). In contrast, CCI and PPR showed no difference between filtered and unfiltered platelets (p = 0.36 and p = 0.29, respectively) because they combined the effects of dose, filtration, and patient size. Slightly fewer patients are required for a study analyzed by regression analysis. CONCLUSION: Regression analysis of posttransfusion platelet increments should be used instead of CCI or PPR to compare the efficacy of platelet components. CCI and PPR should not be used to define platelet refractoriness as a study outcome, because these measures are biased in favor of platelet preparation techniques that provide fewer platelets. PMID- 10378839 TI - FcgammaRIIIB polymorphism: evidence that NA1/NA2 and SH are located in two closely linked loci and that the SH allele is linked to the NA1 allele in the Danish population. AB - BACKGROUND: The neutrophil-specific antigens NA1, NA2, and SH are well-recognized allotypic forms of FcgammaRIIIB. Individuals carrying all three FcgammaRIIIB genes were recently described. STUDY DESIGN AND METHODS: A Danish population (n = 200) was typed for NA1, NA2, and SH by polymerase chain reaction with sequence specific primers (PCR-SSP). Twelve individuals with three FcgammaRIIIB genes were further genotyped by PCR-based restriction fragment length polymorphism and by DNA sequencing. Family studies were performed on three individuals who carry three FcgammaRIIIB genes. RESULTS: The gene frequencies for NA1, NA2, and SH were 0.365, 0.635, and 0.030, respectively. In eight individuals (4%), all three FcgammaRIIIB genes were identified. All 12 SH+ individuals were NA1+. CONCLUSION: The NA1, NA2, and SH gene frequencies observed in Danes are similar to those in other white populations. The distribution of FcgammaFIIIB genotypes in the Danish population strongly indicates that the NA and the SH systems are located in two closely linked loci and that SH is closely linked to NA1. Finally, a new PCR-SSP was developed to distinguish NA2 from SH. PMID- 10378840 TI - Photodynamic sterilization of red cells and its effect on contaminating white cells: viability and mechanism of cell death. AB - BACKGROUND: Phthalocyanines are useful sensitizers for photodynamic sterilization of red cell concentrates. Various lipid-enveloped viruses can be inactivated with only limited red cell damage. Because white cells are involved in the immunomodulatory effects of blood transfusions, the study of the effect of photodynamic treatment on these cells is imperative. STUDY DESIGN AND METHODS: White cell-enriched red cell suspensions were photodynamically treated with either the hydrophobic Pc4 (HOSiPcOSi-(CH3)2(CH2)3N(CH3)2) or water-soluble aluminum phthalocyanine tetrasulfonate (AIPCS4) under virucidal conditions. Viability of white cell subpopulations on Days 0, 1, and 4 after treatment was determined by fluorescence-activated cell sorting by flow cytometric analysis of propidium iodide uptake. Apoptosis induction was studied by DNA ladder formation and staining for an early marker of apoptosis (annexin V). RESULTS: Treatment with Pc4 causes a significant decrease in cell viability of all white cells, as shown by prodidium iodide uptake. Monocytes and granulocytes are the most sensitive, and lymphocytes are relatively more resistant. Some of the cells die by apoptosis, which is induced within 30 minutes after treatment. Treatment with AIPCS4 damages only monocytes; other cell populations are not affected. CONCLUSIONS: Physicochemical properties of the photosensitizers partly determine their effect on white cells. Differences in intracellular localization are likely to be responsible for the effects observed. PMID- 10378841 TI - Blood irradiation for intraoperative autotransfusion in cancer surgery: demonstration of efficient elimination of contaminating tumor cells. AB - BACKGROUND: Intraoperative blood salvage is contraindicated in cancer surgery because of contaminating tumor cells and the risk of systemic dissemination. On the basis of the radiosensitivity of cancer cells, irradiation of salvaged blood with 50 Gy is proposed as a way to allow return of salvaged blood. STUDY DESIGN AND METHODS: Elimination of tumor cells by blood irradiation was studied in vitro with cells from 10 cell lines and from 14 tumor preparations after their addition to red cells in high numbers, or with blood shed during cancer surgery. Before and after gamma radiation, tumor cells were isolated by density gradient centrifugation and tested for their proliferative capacity in a cell colony assay. DNA metabolism was analyzed by incorporation of 5' bromodesoxyuridine. RESULTS: Survival curves of cells from various tumors confirmed D0 (the dose required to reduce the fraction of surviving cells to 37 percent of the original value) values in the range of 1.2 to 2.2 Gy. After irradiation of tumor cell contaminated blood with 50 Gy, no cell colony formation was observed, which indicates a reduction rate exceeding 10 log. Irradiated cancer cells showed viability, but no residual DNA metabolism. CONCLUSION: The level of inactivation by a 50-Gy dose far exceeds that needed to inactivate the number of proliferating tumor cells observed or expected in wound blood. These results provide the experimental basis for the clinical application of blood irradiation for intraoperative blood salvage in cancer surgery. PMID- 10378842 TI - Platelet membrane integrity during storage and activation. AB - BACKGROUND: The platelet cell membrane appears to undergo a lipid-phase transition on cooling from 23 degrees C to 4 degrees C. Consequences of this phase transition are leakage of cellular material and irreversible cellular damage. Whether agents, of known benefit in protecting membranes and proteins from cooling and drying injury, could also protect platelets was investigated. Leakage of cytosolic components was assessed by measuring the release of fluorescein into the surrounding medium. STUDY DESIGN AND METHODS: Fresh platelets were suspended in 5 percent dimethyl sulfoxide (DMSO) or in 5 mM of one the following agents: glucose, trehalose, sucrose, glycerol, ethylene glycol, 1,2 propanediol, or L-proline. Platelets were loaded with 10 nMfluorescein diacetate (FD), chilled at 4 degrees C for 24 hours or frozen at -1 degree C per minute to 70 degrees C, warmed rapidly at 37 degrees C, and centrifuged, and the supernatant was measured for the presence of fluorescein. The effect of FD on platelets was assessed by agglutination with ristocetin, aggregation with thrombin and ADP, platelet-induced clot retraction, and expression of p-selectin. Platelet function and activation before and after freezing or cooling were measured by the same methods. RESULTS: By flow cytometry, 98 percent of the platelets incorporated FD. The trapped fluorescein resulted in neither platelet activation (p = 0.9) nor reduction of platelet function (p = 0.12-0.94) from that in control platelets. Freezing of platelets in DMSO caused far less release of fluorescein than did freezing with other agents (p<0.001) or chilling of platelets at 4 degrees C for 24 hours (p<0.0001). Supernatant levels of fluorescein correlated inversely with platelet function. Fluorescein was also shown to be released during aggregation with thrombin or ADP but not during agglutination with ristocetin. CONCLUSIONS: Release of fluorescein into the surrounding medium indicated a loss of platelet membrane integrity and function. Cellular loading with FD is a simple method of studying membrane integrity of platelets and other cells. PMID- 10378843 TI - Transfusion of buffy coat-depleted blood components and risk of postoperative infection in orthopedic patients. AB - BACKGROUND: Allogeneic blood transfusions have been reported to increase susceptibility to postoperative infection, but the findings were inconclusive. This study was designed to investigate the effect of buffy coat-depleted allogeneic and autologous transfusion on postoperative infection in patients undergoing orthopedic surgery. STUDY DESIGN AND METHODS: Patients (n = 385) undergoing elective orthopedic surgery (primary and revision joint replacement, spinal, or pelvic surgery) were included in a prospective observational study of the incidence of postoperative infection between April and December 1996. Infection rates in patients who received allogeneic buffy coat-depleted blood transfusions were compared with those in patients who received no transfusion or only autologous (buffy coat-depleted) blood. RESULTS: Patients without exposure to allogeneic blood (no blood or only autologous blood) had an infection rate of 3.9 percent, as compared to a rate of 12.2 percent for those with exposure to allogeneic blood (allogeneic blood, autologous plus allogeneic blood) (odds ratio 3.442; 95% CI, 1.349-10.40; p = 0.006). Of the 385 study patients, 309 underwent primary hip or knee replacement surgery. In this homogeneous subgroup, the postoperative infection rate was 4.6 percent after no transfusion or autologous transfusion and 11.9 percent after allogeneic transfusion (odds ratio 2.827; 95% CI 1.059-8.799; p = 0.036). Multivariate regression analysis confirmed buffy coat depleted allogeneic blood transfusion as an independent variable associated with high risk for postoperative infection. CONCLUSION: Buffy coat-depleted allogeneic blood transfusion increases the incidence of postoperative infection in patients undergoing uncontaminated orthopedic surgery. PMID- 10378844 TI - Evaluation of RNA and E2 antibodies in prospectively followed recipients of hepatitis G virus-infected blood. AB - BACKGROUND: Hepatitis G virus (HGV) has recently been cloned and tests for HGV RNA and envelope antibodies (anti-E2) have been developed. HGV infection is widespread among blood donors worldwide, but the clinical and serologic outcome of transfusion-associated HGV infection has not been fully characterized. STUDY DESIGN AND METHODS: Consecutive blood donors (n = 2210) were investigated for HGV markers (RNA and anti-E2). The recipients of HGV RNA-positive blood were followed for 1 year after transfusion. RESULTS: Forty-two blood donors (1.9%) were positive for HGV RNA. Eight recipients of HGV RNA-positive blood were retrospectively identified within 2 weeks of transfusion and prospectively followed. In four patients, the presence of anti-E2 before transfusion or an early antibody response protected them from reinfection or prevented HGV persistence, while, in the remaining four patients, transient or persistent viremia was detected shortly after exposure. None of the infected recipients had any evidence of liver disease. CONCLUSION: These results do not support the screening of donors to prevent transfusion-associated HGV infection. PMID- 10378845 TI - Human T-lymphotropic virus type I among blood donors from Guadeloupe: donation, demographic, and biologic characteristics. AB - BACKGROUND: Epidemiologic data on human T-lymphotropic virus type I (HTLV-I) in Guadeloupe (French West Indies) are scant. STUDY DESIGN AND METHODS: From January 1989 to December 1996, 59,426 blood donors were screened by enzyme immunoassay for antibodies to HTLV-I. All repeatedly reactive samples were confirmed by Western blot. Temporal trends in HTLV-I seropositivity rates were examined during the study period. A multivariate analysis of donation, demographic, and biologic characteristics was performed. RESULTS: Of the screened blood donors, 195 were confirmed as seropositive, for an overall prevalence of 0.33 percent (95% CI 0.28 0.38). A marked decrease in overall HTLV-I prevalence with time (from 0.47% in 1989 to 0.13% in 1996) was observed, which can be explained mainly by the decreasing percentage of recruited new donors during the study period. Four independent risk factors for HTLV-I were identified: new donor status (odds ratio [OR] 12.5), female sex (OR 1.7), increasing age (30-39 years: OR, 2.4; 40-49 years: OR, 3.7; >50 years: OR 6.6), and positive antibodies to hepatitis B virus core antigen (OR, 1.7). Selection of specific locations for blood collection was inversely associated with HTLV-I (OR 0.5). CONCLUSION: New donor status, advancing age, female sex, and positivity for hepatitis B virus core antibodies were the major factors associated with HTLV-I infection in Guadeloupe. PMID- 10378846 TI - Cost of umbilical cord blood units released for transplantation. AB - BACKGROUND: A large number of institutions have started programs banking umbilical cord blood (UCB) for allogeneic unrelated-donor and related-donor transplantation. However, limited information is available on the financial issues surrounding these activities. STUDY DESIGN AND METHODS: The aim of this study was to determine the fee per UCB unit released for transplantation that would allow cost recovery after 10 years. Three organizational models were considered suitable to provide units for five UCB transplants per 1 million population per year, a figure that would translate into an annual need for 280 units in Italy. Models A, B, and C included, respectively, seven networked banks, each with an inventory of 1,500 units; two networked banks, each with an inventory of 5,000 units; and one bank with an inventory of 10,000 units. It was estimated that it would take 3 years to develop the cryopreserved inventory and that approximately 3 percent of the inventory could be released and replaced each year during the 7-year interval between the fourth and tenth years of activity. The data on the costs of labor, reagents and diagnostics, disposables, depreciation and maintenance, laboratory tests, and overhead, as well as the operational data used in the analysis were collected at the Milano Cord Blood Bank in 1996. RESULTS: Fees of US $15,061, $12,666, and $11,602 per unit released during the fourth through the tenth years of activity allow full cost recovery (principle and interest) under Models A, B, and C, respectively. CONCLUSION: Although UCB procurement costs compare favorably with those of other hematopoietic cell sources, these results and the current fee of US $15,300 used in some institutions show that UCB is an expensive resource. Therefore, judicious planning of banking programs with high quality standards is necessary to prevent economic losses. The advantages of lower fees associated with the centralized banking approach of Model C should be balanced with the more flexible collection offered by Model A. PMID- 10378847 TI - A survey of phlebotomy among persons with hemochromatosis. AB - BACKGROUND: One in 10 whites in the United States is a carrier for hemochromatosis and an estimated 1 in 200 is clinically affected. Early treatment with therapeutic phlebotomy to remove excess iron can prevent associated chronic diseases. However, little information is available on the amount of blood withdrawn or the rates of withdrawal from hemochromatosis patients. The patterns of therapeutic phlebotomy and the magnitude of charges in persons with hemochromatosis were surveyed. STUDY DESIGN AND METHODS: Surveys were mailed to persons with hemochromatosis identified by health care providers, blood centers, patient advocacy groups, and the Internet. There were 2362 respondents to the survey from the United States. RESULTS: Thirty-seven percent of respondents reported being voluntary blood donors prior to diagnosis. The mean rate of therapeutic phlebotomy for iron depletion was 2.6 units per month (mean duration, 13 months). The mean rate of maintenance phlebotomy was 0.5 units per month. Therapeutic phlebotomy rates varied by sex, age, reason for diagnosis, and severity of symptoms. Seventy-six percent of respondents reported full or partial insurance coverage of therapeutic phlebotomy charges. Seventy-six percent received therapeutic phlebotomy services in a hospital or physician's office and 30 percent in a blood center. Charges for therapeutic phlebotomy varied by site, with a mean cost of $90 in hospitals and $52 in blood centers. Fifty-four percent of respondents attempted to donate blood after their diagnosis but were excluded. CONCLUSION: The amount of blood withdrawn from persons with hemochromatosis is substantial. The location where patients received phlebotomy services appears to be influenced by charges and time since diagnosis. PMID- 10378848 TI - Red cell transfusion outcome studies require a prospective, randomized design. PMID- 10378849 TI - Effectiveness of a highly sensitive chemiluminescent immunoassay in screening for hepatitis B surface antigen in Japanese blood donors. PMID- 10378850 TI - Collection of mononuclear cells in the Spectra for the generation of dendritic cells. PMID- 10378851 TI - Nomenclature of granulocyte alloantigens. ISBT Working Party on Platelet and Granulocyte Serology, Granulocyte Antigen Working Party. International Society of Blood Transfusion. PMID- 10378853 TI - Optimal management of bleeding and transfusion in patients undergoing cardiac surgery. AB - Patients undergoing cardiac surgery with cardiopulmonary bypass (CPB) are at increased risk for excessive perioperative blood loss requiring transfusion of blood products. Point-of-care evaluation of platelets, coagulation factors, and fibrinogen can enable physicians to rapidly assess bleeding abnormalities, facilitate the optimal administration of pharmacological and transfusion-based therapy, and also identify patients with surgical bleeding. The ability to reduce the unnecessary use of blood products in this setting has important implications for emerging issues in blood inventory and blood costs. The ability to decrease surgical time, along with exploration rates, has important consequences for health care costs in an increasingly managed health care environment. PMID- 10378852 TI - Effects of nicardipine and nitroglycerin on perioperative myocardial ischemia in patients undergoing coronary artery bypass surgery. AB - Perioperative myocardial ischemic episodes are predictive of adverse cardiac outcomes after coronary artery bypass surgery. We compared the efficacy of continuous infusions of nicardipine (group NIC) and nitroglycerin (group NTG) in reducing the frequency and severity of myocardial ischemic episodes. Patients received either a nicardipine infusion, 0.7 to 1.4 microg/kg/min (n = 30), nitroglycerin infusion, 0.5 to 1 microg/kg/min (n = 30), or neither medication (group C; n = 17) after aortic occlusion clamp release and for 24 hours postoperatively. Myocardial ischemic episodes were considered as ST segment depressions or elevations of 1 mm or greater from baseline, each at J + 60 milliseconds and lasting 1 minute or greater, using a two-channel Holter monitor. Only nicardipine significantly decreased the duration (3.2 +/- 1.2 min/h) and the area under the ST time curve (AUC; 5.7 +/- 15.7 AUC/h) of 1-mm or greater myocardial ischemic episodes compared with group C (17.2 +/- 5.6 min/h and 30.1 +/- 49 AUC/h, respectively) during the intraoperative postbypass period. A trend toward lower frequency, duration, and area under the ST time curve of myocardial ischemic episodes was observed in group NIC compared with group NTG. Cardiac indices and mixed venous oxygen saturations were significantly greater, whereas systemic pressures were less in group NIC compared with group NTG for the same period. These results suggest that nicardipine, but not nitroglycerin, decreased the duration and area under the ST time curve of myocardial ischemic episodes shortly after coronary revascularization. Larger studies are required to verify the efficacy of nicardipine in reducing the severity of myocardial ischemia during cardiac surgery. PMID- 10378854 TI - Neurological complications of cardiac surgery: the need for new paradigms in prevention and treatment. AB - Neurological injury is a devastating complication of cardiac surgery that results in a longer duration of hospitalization, increased costs, and increased likelihood of death. Such injury can affect any level of the central nervous system, and its manifestations are broad, ranging from neurocognitive dysfunction to frank stroke. Many variables have been found to be indicative or risk for perioperative neurological injury, but the predictive models are more useful for stroke risk than for neurocognitive dysfunction. Strategies aimed at reducing neurological injury during cardiac surgery have focused, for the most part, on the technical aspects of cardiopulmonary bypass. The concomitant performance of carotid endarterectomy and cardiac surgery continues to be controversial, although the management of patients with symptomatic carotid stenosis is better defined. Cerebral embolism, including atheroembolism from the ascending aorta, has an important role in the pathogenesis of neurological injury of all types. Epiaortic ultrasound imaging of the aorta is a sensitive technique for the identification of atherosclerosis of the ascending aorta at the time of surgery, which can allow it to be avoided and therefore reduce the risk for atheroembolism. Results of laboratory investigations have provided insight into the mechanisms of ischemic neuronal injury and a basis for the development of neuroprotective drugs. Neuroprotection may best be accomplished during cardiac surgery because, in contrast to nonsurgical situations, potential agents can be administered before the neurological insult occurs. Reducing the incidence of perioperative stroke will require a multidisciplinary approach that includes novel diagnostic and therapeutic strategies. PMID- 10378855 TI - Intraoperative considerations during minimal-access cardiac surgery. AB - The range of minimal-access cardiac surgery approaches has many implications in intraoperative management. A modified anesthetic regimen is required to deal with the type of surgical exposure, hemodynamic instability, whether cardiopulmonary bypass is used, and early extubation. Intraoperative considerations include hemodynamic monitoring, one-lung ventilation, pharmacological stabilization of the myocardium, pacing, hypothermia, bleeding, and rapid emergence with a minimum of postoperative mechanical ventilation. As a result, anesthetic methods and intraoperative management were modified to meet these specific needs of minimally invasive cardiac procedures. PMID- 10378856 TI - Prevalence of left ventricular diastolic filling abnormalities in adult cardiac surgical patients: an intraoperative echocardiographic study. AB - The incidence of left ventricular (LV) diastolic abnormalities in adult cardiac surgical patients has not previously been adequately investigated. The present study was performed to characterize LV diastolic filling patterns by performing transesophageal Doppler echocardiographic (TEE) studies in patients undergoing cardiac surgical procedures and thus indirectly assess diastolic function in these patients. Doppler TEE studies were performed and transmitral flow (TMF) and pulmonary venous flow (PVF) velocities were recorded in 104 patients intraoperatively. Peak early (E) and late (A) TMF velocities and systolic (S) and diastolic (D) forward PVF velocities were assessed and deceleration time (DT) was measured in all patients. For analysis, the study patients were classified into three groups according to the ratio of the TMF E to A velocity curves: group I with E/A ratio less than 1.0, group II with E/A ratio of 1.0 to less than 2.0, and group III with E/A ratio of 2.0 or greater. A filling pattern of abnormal LV relaxation was found in 73 patients (E/A < 1.0), a normal or pseudonormal pattern was present in 27 patients (1.0 < or = E/A < or = 2.0), and restrictive filling in 4 patients (E/A > 2.0). Patients with impaired relaxation had a greater incidence of recent myocardial infarction and congestive heart failure (CHF) than those with normal or pseudonormal filling patterns. Within group II, patients with CHF had higher TMF E deceleration rates and lower PVF S/D ratios compared with those without CHF (P < .05). Doppler echocardiographic examination of TMF and PVF velocities suggests that abnormalities in diastolic function are prevalent in adult cardiac surgical patients. PMID- 10378858 TI - Historical perspective: surgery for chronic thromboembolic disease. AB - Obstruction of major pulmonary vessels with organized thromboemboli is a rare sequelae of acute pulmonary embolic disease. Depending on the extent and duration of vascular occlusion, patients experiencing this unusual disorder may develop significant pulmonary hypertension and cor pulmonale. If left untreated, the ultimate clinical outcome is right heart failure and death. Over the past several decades, the description of this clinical entity has evolved from an autopsy curiosity to a recognized cause of chronic pulmonary hypertension. Also, during this same time period, surgical capabilities have greatly advanced, providing these patients a potentially life-saving remedy for this debilitating form of pulmonary vascular disease. This article provides a historical perspective for our current understanding of major vessel chronic thromboembolic pulmonary hypertension as a distinct clinical disorder. It also chronicles the developments in surgical techniques that have made thromboendarterectomy of the pulmonary arterial bed a reality. PMID- 10378857 TI - Echocardiographic characterization of left ventricular diastolic properties in patients presenting for the maze procedure. AB - The aim of this study is to characterize and compare the left ventricular (LV) diastolic filling patterns in patients with paroxysmal (PAF) versus chronic atrial fibrillation (CAF) undergoing the maze procedure and to examine their relation with the hemodynamic status. Fifty patients with PAF and 22 with CAF were studied. Hemodynamic measurements and transesophageal echocardiography (TEE) were performed after the induction of anesthesia but before surgical incision, at stable conditions. Transmitral (TMF) and pulmonary venous flow (PVF) velocities were recorded with the pulsed Doppler method. Statistical analysis between the two groups (PAF and CAF) was performed using Student's t-test and chi-squared test, with P less than .05 statistically significant. Compared with patients in the PAF group, those in the CAF group had: (1) higher pulmonary capillary wedge pressure (14 +/- 5 v 12 +/- 4 mm Hg; P < .05), (2) lower left ventricular fraction of area change (43% +/- 6% v 52% +/- 9%; P < .01), (3) slower PVF systolic wave velocity (23 +/- 10 v 35 +/- 15 cm/s; P < .05), and (4) lower ratio of PVF systolic to diastolic wave velocity (0.75 +/- 0.3 v 1.2 +/- 0.4; P < .05). In the present study, LV filling patterns of abnormal relaxation were found in all our patients who underwent the maze procedure for CAF or PAF. Although the cause of LV filling abnormalities is not apparent, the data suggest LV diastolic dysfunction is prevalent in these patients. PMID- 10378859 TI - The evolution and the current state of the art of pulmonary thromboendarterectomy. AB - Optimal reduction in pulmonary vascular resistance caused by chronic pulmonary embolism is obtained by bilateral pulmonary thromboendarterectomy with removal of occlusive material in all bronchopulmonary segmental arteries that are partially or completely obstructed. The most effective way to obtain this goal is the use of median sternotomy with cardiopulmonary bypass, deep hypothermia, and intermittent periods of circulatory arrest. During circulatory arrest, thromboendarterectomy is performed by specially designed dissectors that allow simultaneous dissection and removal of blood from the surgical field. The operative mortality rate for pulmonary thromboendarterectomy at the University of California, San Diego, between 1990 and 1998 was 9.2% in 1,049 patients. PMID- 10378860 TI - Outcome after pulmonary thromboendarterectomy. AB - Pulmonary thromboendarterectomy (PTE) surgery is an effective treatment for the majority of patients with chronic thromboembolic pulmonary hypertension (CTEPH), with an acceptable mortality rate. The immediate outcome after PTE has repeatedly shown a dramatic decline in pulmonary vascular resistance, echocardiographic evidence of reduced right heart pressure, and increased perfusion on lung scan. Prospective long-term follow-up studies (1 to 4 years post-PTE) have found most patients to be in New York Heart Association class I or II, with significantly improved quality of life and ability to walk, climb stairs, return to work, and perform household tasks. Disease-related symptoms and hospitalizations and/or emergency room visits have also been significantly reduced. These findings indicate an improved functional status and longer life expectancy can be achieved after PTE. PMID- 10378861 TI - Postoperative management of the patient undergoing pulmonary thromboendarterectomy. AB - The postoperative course of the patient undergoing pulmonary thromboendarterectomy poses a unique series of challenges in terms of ventilatory care and hemodynamic management. Experience, cooperation, and interaction are necessary among the various disciplines providing care for these patients during the preoperative, operative, and postoperative phases of care. The purpose of this article is to share the approach necessary for the optimal postoperative care of the patient undergoing thromboendarterectomy, to present the theoretical justification for this care, and to delineate the areas of uncertainty that still exist. PMID- 10378862 TI - Evaluation of patients with suspected chronic thromboembolic pulmonary hypertension. AB - Chronic thromboembolic pulmonary hypertension (CTEPH) can be a difficult diagnosis to establish, typically requiring a high index of suspicion on the part of the clinician when challenged by a patient reporting exertional dyspnea. The importance of this diagnosis is heightened by the appreciation that it is a potentially curable form of pulmonary hypertension, by a surgical procedure called pulmonary thromboendarterectomy. This article highlights the clinical presentation, evaluation, and criteria for surgical candidacy of those patients suspected of having CTEPH. PMID- 10378863 TI - Cell surface expression of MHC molecules in glioma cells infected with herpes simplex virus type-1. AB - 9L glioma cells consistently expressed major histocompatibility complex (MHC) class I but not class II molecules. Herpes simplex type-1 virus (HSV-1) infection significantly reduced the expression of MHC I on the cell surface. Recombinant interferons could enhance the cell-surface expression of MHC I but had no effect on MHC II. This enhancement was partially inhibited by HSV-1 infection. HSV-1 mutants with deletions in ICP4, ICP6, ICP27, ICP47 and UL41 genes do not affect the infection induced inhibition, suggest that a different mechanism may be employed in the inhibition of cell-surface expression of MHC molecules. PMID- 10378865 TI - Neurokinin type-1 receptor antagonist inhibits enhancement of T cell functions by substance P in normal and neuromanipulated capsaicin-treated rats. AB - Substance P (SP) plays a major role in the regulation of the interaction between immune and nervous systems. SP administration stimulates Con A-induced proliferation of spleen and peripheral blood lymphocytes from normal and neonatally capsaicin treated rats, which correlated with enhanced IL-2 production and expression of activation antigens such as IL-2 receptor alpha chain (CD25) and RT1B MHC class II molecule. Moreover, SP markedly increased the percentage of CD5+ and CD4+ T lymphocytes in the peripheral blood of capsaicin-treated rats. Concomitant administration of SP with the non-peptide Neurokinin-1 receptor (NK1R) antagonist SR140333 completely inhibited the SP-mediated augmentation of Con A-induced PBL proliferation and IL-2 production as well as of CD4+ CD25+ and CD4+ RT1B+ T cell numbers in normal and capsaicin-treated rats. SR 140333 also blocked the increased percentage of peripheral blood CD4+ T cells induced by SP in capsaicin-treated rats. PMID- 10378864 TI - Cerebrospinal fluid abnormalities in a phase III trial of Avonex (IFNbeta-1a) for relapsing multiple sclerosis. The Multiple Sclerosis Collaborative Research Group. AB - BACKGROUND AND OBJECTIVE: This report provides results of CSF analyses done in a subset of relapsing remitting MS patients participating in a placebo-controlled, double-blind, phase III clinical trial of IFNbeta-Studies supported by the National Multiple Sclerosis Society (grants RG2019, RG2827),a (Avonex , Biogen). The clinical trial demonstrated that IFNbeta-1a treatment resulted in significantly reduced disability progression, annual relapse rate, and new brain lesions visualized by cranial magnetic resonance imaging. The objectives of the current study were to determine: (a) whether CSF abnormalities in MS patients correlated with disease or MRI characteristics, and (b) effects of IFNbeta-1a therapy on these CSF abnormalities. METHODS: CSF was analyzed from 262 (87%) of the 301 study subjects at entry into the clinical trial, and a second CSF sample was analyzed from 137 of these 262 subjects after 2 years of therapy. CSF cell counts, oligoclonal bands (OCB), IgG index, and free kappa light chains were measured using standard assays. Baseline CSF results were compared with demographic, disease, and MRI parameters. Differences in on-study relapse rate, gadolinium enhancement, and EDSS change according to baseline CSF status was used to determine the predictive value of CSF for subsequent clinical and MRI disease activity. Change in CSF parameters after 104 weeks were used to determine the effects of treatment. RESULTS: (1) At study baseline, 37% of the subjects had abnormal CSF WBC counts, 61% had abnormal levels of CSF free kappa light chains, 84% had abnormal IgG index values, and 90% were positive for OCB. (2) Baseline IgG index, kappa light chains, and OCB showed weakly positive, statistically significant correlations with Gd-enhanced lesion volume and T2 lesion volume. WBC showed a statistically significant correlation with Gd-enhancing lesion volume but was uncorrelated with T2 lesion volume. (3) There was an associated between baseline CSF WBC counts and on-study clinical and MRI disease activity in placebo recipients. (4) IFNbeta-1a treatment resulted in significantly reduced CSF WBC counts, but there was no treatment-related change in CSF IgG index, kappa light chains, or OCB, which remained relatively stable over time in both patient groups. CONCLUSIONS: The current study documents significant reductions in CSF WBC counts in patients treated with IFNbeta-1a for 104 weeks. This finding is considered relevant to the therapeutic response, since CSF WBC counts were found to be positively correlated with subsequent clinical and MRI disease activity in placebo-treated relapsing MS patients. PMID- 10378866 TI - Pathogenesis of acute passive murine encephalomyelitis II. Th1 phenotype of the inducing population is not sufficient to cause disease. AB - The present study was designed to assess the pattern of cytokine expression over the course of disease in the central nervous system (CNS) of recipients of an encephalitogenic T-cell clone specific for proteolipid protein (PLP) peptide 139 151. Reverse transcriptase-polymerase chain reaction (RT-PCR) analyses of CNS mRNA from samples taken during the onset of acute disease demonstrated upregulation of message for cytokines involved in the recruitment and activation of macrophages (GM-CSF, interleukin (IL)-3, IL-9) and the inflammatory cytokines tumor necrosis factor (TNF)-alpha and iNOS as well as message for IL-10 and transforming growth factor (TGF)beta. During the recovery stage message for most cytokines was absent, but during relapse inflammatory cytokine messages were again detectable. Message for the accessory molecules B7-2 and CTLA-4 was observed only on the day of onset of acute experimental allergic encephalomyelitis (EAE) and at relapse. The messages for these molecules were downregulated at the onset of recovery. These results illustrate the dynamic nature of the immune response during the course of EAE, and support a model of disease in which T-cells are involved in the regulation of disease while a nonspecific inflammatory reaction is responsible for the CNS damage observed during EAE. PMID- 10378867 TI - Antibody recognition and RNA binding of a neuronal nuclear autoantigen associated with paraneoplastic neurological syndromes and small cell lung carcinoma. AB - The PLE21/HuC neural protein is an autoantigen for anti-neuronal nuclear autoantibodies (ANNA-1/anti-Hu/Type IIa antibodies) from a patient with paraneoplastic limbic encephalomyelitis and small cell lung carcinoma. This antigen belongs to the Hu/ELAV-like protein family, contains three RNA recognition motifs (RRMs) and has RNA binding capacity. In many autoimmune diseases mediated by autoantibodies, antibodies often interfere with the biological functions of their target antigens. To investigate the influences of the autoantibodies on the biological function of the antigen, we mapped the regions which were required for the antibody recognition and for the RNA binding. Deletion analysis of the antigen revealed that the epitopes for the antibodies were localized in the regions of 12 residues, amino acids 161-172, and eight residues, amino acids 29-38, of the first and second RRMs. It was also shown that the eight residues, amino acids 29-38, and the 10 residues, amino acids 187-194, were required for the RNA binding. Although amino acids 29-38 were necessary for both the antibody recognition and the RNA bindings, pre-incubation of the PLE21 antigen with the antibodies did not inhibit the formation of the complex of PLE21, the antibodies and RNA. Thus, the regions required for the antibody recognition are not identical with those for the RNA binding, and it seems unlikely that the autoantibodies interfere with RNA binding of the antigen. PMID- 10378868 TI - Proinflammatory profile of cytokine production by human monocytes and murine microglia stimulated with beta-amyloid[25-35]. AB - Growing evidence indicates that amyloid (A beta) deposition and phagocyte activation participate in inflammatory reactions in the brain during the course of Alzheimer's disease. To further investigate the effects of A beta-phagocyte interaction, we examined the production of proinflammatory (IL-1beta, IL-6), chemotactic (MIP-1alpha, IP-10) and inhibitory (IL-1Ra, IL-10 and TGFbeta1) cytokines by cultured human monocytes and mouse microglial cells upon stimulation with A beta[25-35]. Northern blot analysis and specific immunoassays demonstrated that A beta[25-35] triggers mRNA expression and release of IL-1beta, IL-1Ra and MIP-1alpha but not of IL-6, IL-10, TGFbeta1 and IP-10 from human monocytes. Similar results were obtained by examining the production of IL-1beta, IL-6 and IL-10 from mouse microglial cells in the same experimental conditions. Taken together, these data indicate that A beta-phagocyte interaction can drive a different response towards cytokine production by monocytes and microglia, with a particular proinflammatory trend, and further support a role for A beta deposition as a triggering factor of inflammatory events in Alzheimer's disease. PMID- 10378870 TI - Induction of pro-inflammatory cytokine mRNAs in the brain after peripheral injection of subseptic doses of lipopolysaccharide in the rat. AB - Although it is generally accepted that pro-inflammatory cytokines produced by cells of the central nervous system play important roles in the communication between the central nervous system and the immune system during sepsis, it is not clear whether these cytokines are produced in the brain under subseptic conditions. In this study, we used in situ hybridization to examine the mRNA expression of the pro-inflammatory cytokines IL-1beta and TNFalpha in the brains of rats 2 and 12 h after they were challenged by peripheral injections of lipopolysaccharide (LPS) ranging from 0.01 to 1000 microg/kg. Unlike septic doses of LPS (> 500 microg/kg), which induce global expression of pro-inflammatory cytokines in the brain, subseptic doses of LPS (0.01-10 microg/kg) induced IL 1beta and TNFalpha mRNA expression only in the choroid plexus, the circumventricular organs, and meninges. The expression of the cytokine-responsive immediate early gene I kappaB alpha was induced in the brain after doses of LPS as low as 0.1 microg/kg. I kappaB alpha mRNA expression was confined to sites where IL-1beta and TNFalpha were expressed. These results indicate that the induction and action of pro-inflammatory cytokines during subseptic infection occur at the blood-brain barrier and at circumventricular organs, which may be sites for elaboration of signal molecules that communicate peripheral immune status to the brain. PMID- 10378869 TI - Anti-death properties of TNF against metabolic poisoning: mitochondrial stabilization by MnSOD. AB - The cytokine tumor necrosis factor (TNF) is toxic to some mitotic cells, but protects cultured neurons from a variety of insults by mechanisms that are unclear. Pretreatment of neurons or astrocytes with TNF caused significant increases in MnSOD activity, and also significantly attenuated 3-nitropropionic acid (3-NP) induced superoxide accumulation and loss of mitochondrial transmembrane potential. In oligodendrocytes, however, MnSOD activity was not increased, and 3-NP toxicity was unaffected by TNF. Genetically engineered PC6 cells that overexpress MnSOD also were resistant to 3-NP-induced damage. TNF pretreatment and MnSOD overexpression prevented 3-NP induced apoptosis, and shifted the mode of death from necrosis to apoptosis in response to high levels of 3-NP. Mitochondria isolated from either MnSOD overexpressing PC6 cells or TNF treated neurons maintained resistance to 3-NP-induced loss of transmembrane potential and calcium homeostasis, and showed attenuated release of caspase activators. Overall, these results indicate that MnSOD activity directly stabilizes mitochondrial transmembrane potential and calcium buffering ability, thereby increasing the threshold for lethal injury. Additional studies showed that levels of oxidative stress and striatal lesion size following 3-NP administration in vivo are increased in mice lacking TNF receptors. PMID- 10378871 TI - Antigen presenting capacity of brain microvasculature in altered peptide ligand modulation of experimental allergic encephalomyelitis. AB - Co-immunization with an altered peptide ligand (LR) partially protects SJL mice from proteolipid protein peptide 139-151-induced experimental allergic encephalomyelitis [Kuchroo, V.K., Greer, J.M., Kaul, D., Ishioka, G.Y., Franco, A., Sette, A., Sobel, R.A., Lees, M.B., 1994. A single TCR antagonist peptide inhibits experimental allergic encephalomyelitis mediated by a diverse T cell repertoire. J. Immunol. 153, 3326-3336; Santambrogio, L., Lees, M.B., Sobel, R.A., 1998. Altered peptide ligand modulation of experimental allergic encephalomyelitis: immune responses within the CNS. J. Neuroimmunol. 81, 1-13]. Clinical protection was noted despite extensive central nervous system inflammation observed after co-immunization with native and altered peptides. To extend our previous reports on this model, we now compare MHC class II expression and antigen presenting cell activity of cells associated with the blood-brain barrier in diseased and protected mice. Immunohistochemical studies identified MHC class II products on both the endothelial and microglial/macrophage populations. Ex vivo experiments suggested a correlation between the reduced clinical disease observed in the co-immunized mice and the antigen presenting activity of cells at the blood-brain barrier. The results suggest that antigen presenting activity is primarily mediated by macrophage-lineage cells of the central nervous system. PMID- 10378872 TI - Myelin basic protein-specific T lymphocytes induce chronic relapsing experimental autoimmune encephalomyelitis in lymphocyte-deficient (SCID) mice. AB - Myelin basic protein (BP)-specific T lymphocyte cell lines were selected from the lymph nodes (LN) of BP-immunized, H-2d, CXJ-1 mice prior to the onset of clinical disease. These CD4+ T cells induced severe acute experimental autoimmune encephalomyelitis (EAE) in MHC-compatible (H-2d), lymphocyte-deficient (SCID) mice (C.B-17scid/scid). The incidence of disease was much higher in immunodeficient SCID mice (71%) than in syngeneic immunocompetent CXJ-1 mice (5%). SCID mice with EAE had an acute progressive paralytic disease with inflammation and myelin loss detected in the spinal cord. Eighty-six percent (12/14) of mice followed for more than 2 weeks had 1 or more relapses of EAE. These results demonstrate that clinical remission and relapse of EAE can be induced by the single adoptive transfer of a LN-derived BP-specific T cell line in the absence of host-derived effector and regulatory lymphocytes. Furthermore, the data demonstrate that the pathogenic potential of BP-specific T cells is greater in lymphocyte-deficient SCID mice compared with immunocompetent mice, suggesting that autoreactive T cells are controlled by potent inhibitory mechanisms associated with regulatory lymphocytes. These results are relevant to mechanisms of disease remission and relapse mediated by lymphocytes involved in paralytic inflammatory diseases such as multiple sclerosis (MS). PMID- 10378873 TI - Differential effects of iron load on basal and interferon-gamma plus lipopolysaccharide enhance anticryptococcal activity by the murine microglial cell line BV-2. AB - Here we evaluated the influence of intracellular iron levels on the constitutive and interferon (IFN)-gamma plus lipopolysaccharide (LPS) enhanced anticryptococcal activity by the murine microglial cell line BV-2. We demonstrated that iron loading via ferric nitrilotriacetate (FeNTA) resulted in a significant increase in the constitutive levels of anticryptococcal activity, while the enhancing effects by IFN-gamma plus LPS were prevented. Accordingly, a major increase was observed in the levels of thiobarbituric reactive substance (TBARS) produced upon iron loading under basal conditions, whereas IFN-gamma plus LPS treatment, that per se did not affect TBARS production, prevented by about 50% the enhancement otherwise occurring in response to iron loading. The potential involvement of multiple effector system and their relation to intracellular iron will be discussed. PMID- 10378874 TI - Subcutaneous administration of T-epitope sequences of the acetylcholine receptor prevents experimental myasthenia gravis. AB - Immunization with acetylcholine receptor (AChR) causes experimental myasthenia gravis (EMG). The s.c. administration to C57B1/6 mice of synthetic AChR CD4+ epitopes, before and during AChR immunization, reduced the epitope-specific CD4+ responses and the anti-AChR Ab synthesis, and prevented EMG. The s.c. administration of solubilized AChR had effects similar to those of peptide treatment. Sham-tolerized mice had only Th1 anti-AChR cells, whereas peptide treated mice had also Th2 cells, and Th2-induced anti-peptide Ab. Established EMG was not affected by s.c. peptide treatment, whereas it worsened after s.c. administration of solubilized AChR. PMID- 10378875 TI - Fas (APO-1/CD95) in inflammatory CNS diseases: intrathecal release in bacterial meningitis. AB - Release of Fas (APO-1, CD95), a type L-membrane protein which plays a crucial role in cytokine-mediated apoptosis was investigated in bacterial meningitis, viral meningoencephalitis and multiple sclerosis in vivo. After correction for bloodbrain-CSF-disruption, significantly increased intrathecal release of Fas was demonstrated exclusively in bacterial meningitis arguing for an apoptotic cell death of granulocytes in the subarachnoidal space aimed to self-limit inflammatory host response. PMID- 10378876 TI - Regulation of VIP production and secretion by murine lymphocytes. AB - Vasoactive intestinal peptide (VIP) is a neuropeptide present in the lymphoid microenvironment with a multiplicity of actions. Two sources for VIP have been described in the immune system, the terminals present in central and peripheral lymphoid organs and the immune cells. Although VIP is synthesized by lymphocytes, there is no evidence demonstrating that VIP is released, and which stimuli are able to induce VIP production and secretion. In this study, we demonstrated for the first time, that agents that mediate important immune functions, such as proliferation and antigenic stimulation (Con A, LPS, and anti-TCR antibody), inflammation (LPS, TNFalpha, IL-6 and IL-1beta) or apoptosis (dexamethasone) induce the production and release of VIP to the lymphoid microenvironment. We conclude that VIP is produced and secreted by lymphocytes and propose that during an immune response, the timely release of VIP within the lymphoid organs and peritoneum should influence the differentiation and/or downregulation of the ongoing response. PMID- 10378877 TI - Increased interleukin-6 expression by microglia from brain of aged mice. AB - Over expression of inflammatory cytokines in the brain may establish a state that is permissive to the onset of neurodegenerative disease. Because the occurrence of certain neurodegenerative diseases increases with age, in the present study we examined the expression of the inflammatory cytokine, interleukin-6 (IL-6), in the brain of aged mice. In an initial experiment, IL-6 was measured in crude protein extracts from brains of juvenile (1-month-old), adult (3-month-old), and aged (24-month-old) male BALB/c mice. The concentration of IL-6 in crude protein extracts from the cerebellum, cerebral cortex, and hippocampus increased with age. The increase in IL-6 was discrete, as levels in the hypothalamus were not age-dependent. To begin evaluating spontaneous IL-6 production in aging, glial cells were cultured from brains of neonate, adult, and aged mice. An age associated increase in IL-6 mRNA and supernatant IL-6 concentration was evident, indicating glia from aged mice spontaneously express high levels of IL-6 relative to glia from adult and neonate mice. Flow cytometric analysis revealed that cultures established from aged brain compared to either adult or neonate brain comprised more microglia (i.e., MAC-1-positive cells). Furthermore, the proportion of microglia that was positive for IL-6 increased with age, whereas the proportion of astrocytes that were positive for IL-6 was not age-dependent. The present results suggest that IL-6 increases in the mouse brain with age, and that microglia cultured from aged mice spontaneously produce more IL-6 than those from neonate or adult mice. Therefore, microglia may contribute to the increased level of IL-6 present in aged brain. PMID- 10378878 TI - Synergistic interaction of catecholamine hormones and Mycobacterium avium results in the induction of interleukin-10 mRNA expression by murine peritoneal macrophages. AB - The results of this investigation provides evidence that catecholamine hormones interact with macrophages that are infected with Mycobacterium avium resulting in the induction of IL-10 mRNA and protein. The effect of catecholamine hormones was prevented by treating the cells with the beta-adrenergic receptor antagonist propranolol but not by alpha-adrenergic antagonist phentolamine. The effect of catecholamine stimulation was mimicked by the addition of beta-2 adrenergic agonists and by the addition of cAMP to the infected macrophage cultures. These observations suggest that sympathetic nervous system activation together with microbial infection results in a synergistic interaction that could result in the control of inflammatory processes. PMID- 10378880 TI - Differential immune responses in mice with left- and right-turning preference. AB - Humoral and cell-mediated immune responses of inbred BALB/c male mice were assayed for differential reactivities associated with behavioral sidedness, which was evaluated by spontaneous rotational behavior in a circular cage model system. Mice with left-turning preference had lower in vivo primary IgM and IgG anti Keyhole Limpet Hemocyanin (KLH) antibody responses, delayed-type hypersensitivity (DTH) responses, and host-resistance against the intracellular bacteria, Listeria monocytogenes, than mice with right-turning preference. The only immune parameter not shown to be associated with turning preference was the secondary humoral immune response to KLH. The weak innate immune response of left-turners for clearance of Listeria showed close intercorrelation with elevated serum IL-6 levels. Serum corticosterone and splenic norepinephrine levels were differentially increased and decreased by infection, respectively. We suggest that the observed differential immune reactivities of individual animals with same age, gender, and genetic background are associated with functional asymmetries within the brain, that the hypothalamic-pituitary-adrenal (HPA) axis and sympathetic innervation are involved in the regulatory brain: immune interconnection after infection, and that the HPA axis and sympathetic nervous system are involved in the brain laterality effects on immune responses. PMID- 10378879 TI - CP-10, a chemotactic peptide, is expressed in lesions of experimental autoimmune encephalomyelitis, neuritis, uveitis and in C6 gliomas. AB - CP-10 (chemotactic protein of m.w. 10,000) is a member of the S100 superfamily of Ca2+ binding peptides, which has potent chemotactic activity for murine and human myeloid cells. Here we report on the generation of monoclonal antibodies against CP-10 and accumulation of CP-10+ cells during experimental autoimmune encephalomyelitis (EAE), neuritis (EAN), uveitis (EAU) and in experimentally transplanted C6 gliomas. During acute inflammation, CP-10 is mainly expressed by large ED1+ monocytic perivascular cells that accumulate at days 11-14. CP-10+ cells are predominantly located in areas of cellular infiltration but are as well found in the meninges and infiltrating the brain parenchyma. In transplanted gliomas, CP-10+ cells are located exclusively within the tumor parenchyma. Using double labeling experiments, other cells participating in the inflammatory reaction were found to express CP-10, like few lymphoblastic W3/13+ cells in the vicinity of the inflammatory infiltrate. PMID- 10378881 TI - Identification of a T cell chemotactic factor in the cerebrospinal fluid of HIV-1 infected individuals as interferon-gamma inducible protein 10. AB - Central nervous system (CNS) involvement is a prominent feature of human immunodeficiency virus (HIV-1) infection. Monocytes and CD4+ T cells traverse the blood brain barrier (BBB), and serve as vehicles for the virus and perpetrators for brain pathology by their production of neurotoxins. In the present study cerebrospinal fluid (CSF) samples from HIV-1-infected patients were analyzed for the presence of chemotactic factors. All 36 CSF samples from the patients were positive for the CXC chemokine interferon-gamma inducible protein (IP-10), which was not detected in CSF samples of 14 controls. The IP-10 concentrations were higher in HIV-1-infected patients with HIV-1 associated neurologic disorders than in those without neurological deficits. In contrast to IP-10, other chemotactic factors including the CC chemokines MCP-1, MIP-1alpha, MIP-1beta and RANTES and the cytokines IL-15 and IL-16 were either not detected or increased in only less than 30% of the patients. Unlike the CSF samples of controls, all CSF samples from HIV-1-infected patients induced chemotaxis of T cells activated with IL-2. The significance of IP-10 as a T cell chemotactic cytokine in HIV-1-infected CSF is shown by (1) the correlation of the IP-10 levels with the extent of T cell chemotaxis, (2) the neutralization of T cell chemotaxis by anti-IP-10 antibodies and (3) the correlation of the chemotactic response of CSF samples on activated T cells and the CSF white cell count in the patients. Our data provide evidence that IP-10 contributes to the accumulation of activated T cells in the CSF compartment in HIV-1-infected individuals. PMID- 10378882 TI - A central role for astrocytes in the inflammatory response to beta-amyloid; chemokines, cytokines and reactive oxygen species are produced. AB - Alzheimer's disease (AD) is the commonest form of adult onset dementia and is characterised neuropathologically by the accumulation of plaques containing beta amyloid (A beta) fibrils, reactive astrocytes, activated microglia, and leukocytes. A beta plays a role in the pathology of AD by directly causing neuronal cytotoxicity and stimulating microglia to secrete cytokines and reactive oxygen species (ROS) which also damage neurons. Here, we demonstrate that A beta activates astrocytes and oligodendrocytes (the most common cell types in the brain) to produce chemokines, in particular MCP-1 and RANTES, which serve as potent in vitro microglial and macrophage chemoattractants. Furthermore, we have shown that A beta activates astrocytes to upregulate pro-inflammatory cytokine expression and enhances the production of ROS. We propose therefore that A beta mediated astrocyte activation initiates an inflammatory cascade which could be targeted for therapeutic intervention in AD. PMID- 10378883 TI - Expression and localization of p80 and p68 interleukin-1 receptor proteins in the brain of adult mice. AB - The biological effects of interleukin-1 (IL-1) are mediated by two distinct receptors, the p80 type I IL-1 and p68 type II IL-1 receptor proteins (IL-1RI and IL-1RII, respectively), both of which have been recently co-localized to the growth hormone synthesizing cells of the adenohypophysis. Previous studies have shown that IL-1 can bind to specific structures in the central nervous system, but the distribution of IL-1RI and IL-1RII proteins in the adult mouse brain has not been reported. Here we have used immunohistochemistry to study the expression, distribution and cellular localization of both isoforms of the IL-1 receptor proteins in the adult mouse brain. Using a combination of processing techniques (AMeX fixation and cryosectioning), we have immunolabeled brain sections for each isoform of the IL-1R. Both isoforms are expressed in the CNS, particularly in neuronal soma of the granular layer of the dentate gyrus and pyramidal cells of fields CA1-CA4 of Ammon's horn of the hippocampus, in epithelial cells of the choroid plexus and ependymal layer, and in neuronal soma of Purkinje cells of the cerebellum. The IL-1RII isoform, but not IL-1RI, is expressed in specific neuronal soma and proximal cell processes of neurons of the paraventricular gray matter of the hypothalamus. These immunohistochemical data directly demonstrate the neuronal expression of both IL-1R proteins in situ. The distribution and cellular localization of IL-1R proteins in the CNS provide a molecular basis for understanding reciprocal interactions between the immune system and the brain. PMID- 10378884 TI - T-cell tumor necrosis factor-alpha receptor binding in myasthenic patients. AB - Myasthenia gravis (MG) is a T-cell-dependent and antibody-mediated autoimmune disease of the neuromuscular junction, in which the cytokine network may be deranged. Specific receptors for tumor necrosis factor (TNF)-alpha, a cytokine with several effects on the neuroimmune system, were found on human lymphocytes. In the present study, we assayed TNF-alpha binding on peripheral blood T-cells from MG patients, finding that T-cells from patients have significantly more TNF alpha receptors than those from controls (Bmax: 654 +/- 12 vs. 133 +/- 4 (mean +/ SEM) receptors/cell). Such TNF-alpha binding sites are of the same type in patients and healthy subjects (Kd: 68.7 +/- 4.3 vs. 70.1 +/- 4.8 (mean +/- SEM) pM). The enhanced T-cell TNF-alpha binding is due to an increased number of TNF alpha receptors on T-helper lymphocytes. These results are discussed in terms of MG immunopathogenesis, since it has been reported that activated T-cells have increased amounts of TNF-alpha receptors. PMID- 10378885 TI - In the absence of T cells, natural killer cells protect from mortality due to HSV 1 encephalitis. AB - The importance of natural killer (NK) cells in the resistance to herpes simplex virus type 1 (HSV-1), a common infection of immunocompromised patients, is unclear. Previous data on the role of NK cells in murine HSV-1 infection has been contradictory. Adoptive transfer studies suggested that NK cells mediated resistance to HSV-1, but in vivo depletion approaches demonstrated that NK cells were not important. We studied the course of HSV-1 infection after intranasal (i.n.) inoculation of E26 mice (lacking NK and T cells), T cell knockout (T cell ko) mice (lacking T cells only), or normal control mice. The E26 mice showed greater mortality and an impaired ability to clear virus from lung and brain compared to T cell ko mice and control mice, and had severe necrotizing HSV-1 encephalitis. Therefore, the data support the hypothesis that NK cells play an important role in the natural defense of murine HSV-1 infection. PMID- 10378886 TI - The real CD34+ events: simplicity versus accuracy and flexibility. PMID- 10378887 TI - CD44 isoforms in normal and leukemic hematopoiesis. PMID- 10378888 TI - CD34+ cells from mobilized peripheral blood retain fetal bone marrow repopulating capacity within the Thy-1+ subset following cell division ex vivo. AB - Ex vivo cell cycling of hematopoietic stem cells (HSC), a subset of primitive hematopoietic progenitors (PHP) with engrafting capacity, is required for transduction with retroviral vectors and to increase transplantable HSC numbers. However, induction of division of HSC ex vivo also may lead to differentiation and loss of in vivo marrow repopulating potential. We evaluated mobilized peripheral blood (MPB) PHP for maintenance of stem cell function after ex vivo culture under conditions that we show can induce cycling of a majority of PHP with minimal differentiation. The following methods were combined: cell labeling with the division tracking dye carboxyfluorescein-diacetate succinimidylester (CFSE), analysis of primitive cell surface marker expression, an ex vivo PHP assay, and an in vivo marrow repopulating assay. MPB-purified CD34+ Thy-1+ cells were labeled with CFSE dye and cultured for 112 hours in serum-deprived medium in the presence of the cytokine combinations of thrombopoietin (TPO), flt3 ligand (FL), and c-kit ligand (KL), or TPO, FL, and interleukin 6 (IL-6). Both cytokine combinations supported division of greater than 95% of cells within 112 hours with an average 2.1-fold (TPO, FL, KL) or 1.3-fold (TPO, FL, IL-6) increase in total cell numbers. An average of 21.6% (TPO, FL, KL) and 27.4% (TPO, FL, IL-6) of the divided cells still expressed the Thy-1 marker after 112 hours. Functional assays were performed to compare cultured and uncultured cells. CD34+ Thy-1+ CFSElo (post division) cells showed maintenance of cobblestone area-forming cell (CAFC) frequency (a mean of 1/9.0) relative to the starting population of uncultured CD34+ Thy-1+ cells (a mean of 1/8.4). In contrast, CD34+ cells that had lost Thy-1 expression during culture (CD34+ Thy-1 CFSElo) showed a mean 5.8 fold reduction in CAFC frequency (a mean of 1/52.5). Only the Thy-1-expressing fraction of cells post culture could engraft in vivo in the SCID-hu bone assay. Because the majority of HSC functional activity post culture was found in the CD34+ Thy-1+ fraction, we focused on this fraction for subsequent analysis. CFSE labeling allows segregation and purification by flow cytometry of cells having undergone discrete numbers of divisions during culture. Very few cells that divided more than four times in culture still expressed Thy-1. Cells that retained expression of Thy-1 during culture retained CAFC activity relative to fresh CD34+ Thy-1+ cells, after undergoing at least two divisions. CAFC frequency decreased after four divisions in culture with TPO, FL, and KL or after three divisions in TPO, FL, and IL-6. We then compared populations of Thy-1+ cells that had undergone sequential numbers of divisions in culture for their ability to engraft in the SCID-hu bone assay. Engrafting ability was retained throughout four divisions in both cytokine combinations. These data demonstrate that primitive MPB CD34+ cells maintain HSC function coincident with Thy-1 expression while undergoing two to four divisions under these culture conditions. Essentially all CD34+ Thy-1+ cells divided under the conditions tested, promoting susceptibility to retroviral transduction. PMID- 10378889 TI - Alteration of p16 (CDKN2) gene is associated with interleukin-2-induced tumor cell growth in adult T-cell leukemia. AB - Because tumorigenesis frequently involves the dysfunction of cell cycle-related proteins, we examined the effect of mutations in CDK inhibitor p16 and its linked genomic loci p15, cl.B, and 1063.7 on the growth of primary adult T-cell leukemia (ATL) cells. Southern blot analysis of primary ATL cells showed a significantly higher incidence of p16 gene alteration in acute ATL than in chronic ATL [67.7% (23/34) vs. 26.1% (6/23), respectively; p<0.003]. Similarly, polymerase chain reaction (PCR) analysis of p16 exon 2 revealed a higher incidence of alteration in acute ATL than in chronic ATL [52.9% (18/34) vs. 26.1% (6/23), respectively; p<0.05]. PCR-single strand conformation polymorphism analysis of exons 1 and 2 of p16 showed no mutations in the patients, with normal pattern by Southern blotting or PCR analysis. Notably five of six chronic ATL patients with abnormal p16 genes progressed to acute crisis within 4 months. PCR analysis of the p16 linked loci 1063.7, p15 exon 2, and cl.B found homozygous deletion in 55.9%, 20.6%, and 2.9% of acute ATL cells and 39.1%, 13.0%, and 0% of chronic ATL cells, respectively, showing no relationship of homozygous deletion in either loci with disease subtypes. In most cases, deletions were seen in multiple genes, including p16. Acute ATL cells had a higher frequency of multigene deletions than chronic ATL cells [44.1% vs. 17.4%; p<0.05]. When leukemic cells were analyzed for interleukin 2 (IL-2) responsive growth, only p16 gene alteration was directly associated with leukemic cell growth activity. Among leukemic cells showing high IL-2 responsiveness, 73.1% (19/26) had p16 gene alteration vs. 27.8% (5/18) of leukemic cells that showed low IL-2 responsiveness (p<0.005). p16 gene alteration was found in 73.3% (14/19) of leukemic cells showing high autonomous growth rates but in only 40.0% (10/25) of those leukemic cells showing low autonomous growth (p<0.03). These results suggest the following: alteration of p16-related genomic regions in ATL is usually a wide rearrangement including the p16 gene; within this region, only p16 gene alteration is associated with disease aggressiveness; and p16 gene deletion may be a proximate event in leukemogenesis. PMID- 10378890 TI - IL-3-induced coexpression of histidine decarboxylase, IL-4 and IL-6 mRNA by murine basophil precursors. AB - Murine low-density bone marrow cells sorted from the blast cell window on the basis of high rhodamine-123 retention (Rh-bright), are highly enriched in histamine-, IL-4-, and IL-6-producing cells. We established by in situ hybridization that up to 50% of this population (around 0.25% of the whole bone marrow) coexpressed the transcripts for these molecules upon stimulation with 1L 3. Rh-bright cells were also positive for mRNA encoding the alpha, beta, and gamma chains of the Fc(epsilon)RI which was functional since aggregated IgE induced the same percentage of cells hybridizing with the HDC probe as IL-3. Clonogenic progenitors and histamine- and cytokine-producing cells copurified in the Rh-bright population, but could be distinguished by their c-kit expression, CFU-C being more frequent in the c-kit(high) fraction, while histamine and IL-6 producers were enriched in the kit(low) counterpart. Ultrastructural analysis of Rh-bright cells revealed essentially two subsets, namely undifferentiated blast cells and basophil precursors. No other lineage-committed population was enriched by this sorting procedure, and it can therefore be concluded that coexpression of HDC, IL-6, and IL-4 transcripts in response to IL-3 or aggregated IgE takes place mainly in hematopoietic precursors belonging to the basophil lineage. PMID- 10378891 TI - Thrombopoietin, flt3, and kit ligands together suppress apoptosis of human mobilized CD34+ cells and recruit primitive CD34+ Thy-1+ cells into rapid division. AB - Various combinations of cytokines have profoundly different effects on inhibition of apoptosis and stimulation of self-renewal division of hematopoietic stem cells (HSC) in short-term, ex vivo culture. Our goal was to quantitate expansion of cells with a primitive CD34+ Thy-1+ phenotype, as well as cell cycling, division history, differentiation, and apoptosis of CD34+ cells enriched from normal donor mobilized peripheral blood (MPB) cells. The balance of these parameters determines the net number of transplantable HSC produced in ex vivo cultures. Comparing several different combinations of cytokines added to 90-hour cultures of MPB CD34 cells, thrombopoietin (TPO), flt3 ligand (FL), and c-kit ligand (KL) gave the best result, with the lowest percentage of apoptotic cells and a mean 1.2-fold increase in the number of CD34+ Thy-1+ cells. A combination of interleukin 3 (IL-3), interleukin 6 (IL-6), and leukemia inhibitory factor (LIF) gave the worst outcome, including a decrease of CD34+ Thy-1+ cell number to a mean of 30% of the starting cell number. Cell division history was tracked using the dye 5-(and 6-) carboxyfluorescein diacetate succinimidyl ester (CFSE). Division of CD34+ Thy-1+ cells was faster and more synchronous in TPO, FL, and KL than in IL-3, IL-6, and LIF, which left a significant proportion of CD34+ cells undivided. Such detailed analyses of short-term, ex vivo cultures generated "replication scores," which allowed prediction of a sixfold improvement of the efficiency of gene transduction of primitive hematopoietic progenitors from MPB, using TPO, FL, and KL to replace IL-3, IL-6, and LIF. Analysis of retroviral transduction efficiency confirmed the increase of transgene expression from MPB primitive hematopoietic progenitors assayed after stromal culture was fivefold, validating the usefulness of multiparameter analysis of short-term cultures for survival and replication of CD34+ Thy-1+ cells. PMID- 10378892 TI - Administration of Flk2/Flt3 ligand induces expansion of human high-proliferative potential colony-forming cells in the SCID-hu mouse. AB - The effects of Flk2/Flt3 ligand (FL) administration on human hematopoiesis were investigated using SCID-hu mice transplanted with human fetal bone fragments. Treatment with recombinant human FL induced significant increases in the frequencies of the high-proliferative potential colony-forming cells and low proliferative potential colony-forming cells in steady-state human bone marrow. FL also promoted the expansion of high-proliferative potential colony-forming cells and low-proliferative potential colony-forming cells in the human bone marrow during the recovery phase after irradiation, which was evident in increases in the frequencies as well as in the absolute numbers of colony-forming cells. Furthermore, higher percentages of CD33+ CD15- cells were found in the marrows treated with FL as compared to that of controls, indicating that FL hastened the recovery of at least some aspect of myelopoiesis after irradiation. These results indicate that FL induces the expansion of primitive hematopoietic progenitor cells in vivo and, therefore, may be useful in treating patients to promote an early hematopoietic recovery after cytoablative therapies. PMID- 10378893 TI - SH2-containing protein tyrosine phosphatases SHP-1 and SHP-2 are dramatically increased at the protein level in neutrophils from patients with severe congenital neutropenia (Kostmann's syndrome). AB - Severe congenital neutropenia (SCN) or Kostmann's syndrome is characterized by a stop in differentiation of myeloid progenitor cells at the myelocytic or promyelocytic stage. The pathophysiology of SCN is still unclear. We previously showed that the tyrosine kinase JAK2 is phosphorylated and activated in neutrophils from patients with severe congential neutropenia. We investigated the role of tyrosine phosphatases in this disease. Expression of the SH2 domain containing tyrosine phosphatases SHP-1 and SHP-2 was analyzed in myeloid cells from patients with SCN in comparison to healthy donors. We investigated tyrosine phosphatase expression in myeloid cells at the protein level by Western blot analysis using polyclonal antisera against SHP-1 and SHP-2. Whereas SHP-1 and SHP 2 were hardly detectable in neutrophils from healthy donors, neutrophils from patients with SCN revealed high amounts of these two proteins in Western blot analyses. Reverse transcriptase-polymerase chain reaction and Northern blot analyses demonstrated no dramatic differences of SHP-1 mRNA in neutrophils from congenital neutropenia patients as compared to healthy donors. SHP-2 mRNA was hardly detectable in the neutrophils from patients and in normal neutrophils. Increased expression of SHP protein correlated with elevated activity of both SHP 1 and SHP-2 in neutrophils of patients with SCN. Taken together, these data indicate differential regulation for SHP-1 and SHP-2 at the protein level in neutrophils from SCN patients in comparison to healthy donors. We suggest that overexpression of SHP-1 and SHP-2 protein in neutrophils and not in mononuclear cells from patients with SCN might be related to the disease, e.g., by defective dephosphorylation of proteins involved in intracellular signaling pathways. PMID- 10378894 TI - Characterization of the stage in natural killer cell development in 14.5-day mouse fetal liver using adult bone marrow stroma. AB - Nonstimulated fetal liver (FL) from 14.5-day gestation mice had no natural killer (NK) cell activity and <3% expressed NK1.1. Even after short-term (3-4 day) culture of FL with the late-acting cytokines, interleukin (IL)-15 or IL-2, little or no NK activity was detected. However, longer-term (13 day) culture with IL-2 plus stroma derived from bone marrow (BM) of adult mice, resulted in extensive proliferation and differentiation to mature NK cells. Cell numbers began to increase after 4 days, and by day 13, they had increased 40-fold and 69% of the cells were NK1.1+ with high NK activity and 5%-10% were NK1.1- B220+. With stroma, but no IL-2, equivalent proliferation occurred, but differentiated cells were predominantly NK1.1- B220+, not NK cells. Culture for 13 days without stroma, but with either IL-2, IL-15, FLTK3-ligand (L) or stroma-conditioned medium, resulted in less than fivefold expansion, and minimal NK activity. Culture with combinations of FLTK3-L or ckit-L plus IL-15 or IL-2 increased both cell number and NK activity, but the increase in cell number was less than that seen with stroma plus IL-2. By limiting dilution assay on stroma plus IL-2, the precursor frequency was 1/(2660+/-292) whole FL cells and the absolute number, but not the frequency, increased during culture on stroma without IL-2. The NK cell progenitors were found in sorted NK1.1- and Sca-1+ c-kit+ lineage- subpopulations at a frequency of 1/(156+/-52.5). Together, these data suggest that the NK lineage cells in FL are primarily in early stages of development. They are highly proliferative, respond to early acting cytokines and express stem cell markers. PMID- 10378895 TI - Downregulation of Wilms' tumor gene (WT1) is not a prerequisite for erythroid or megakaryocytic differentiation of the leukemic cell line K562. AB - The Wilms' tumor gene (WT1) encodes a transcription factor of the zinc finger type. A high expression of WT1 has been detected in a range of acute leukemias, and WT1 is downregulated during induced differentiation of some leukemic cell lines. Overexpression of WT1 in some myeloid cell lines confers resistance to differentiation induction. These observations suggest that a high WT1 expression in hematopoietic cells is incompatible with differentiation. In this study, each of the four different isoforms of WT1 was constitutively overexpressed in the leukemic cell line K562. K562 cells express endogenous WT1, which is downregulated as a response to induced differentiation along the erythroid and megakaryocytic pathways. We now demonstrate that a forced exogenous expression of the four different isoforms of WT1 in K562 does not affect the differentiation response, as judged by accumulation of hemoglobin in response to hemin or the expression of megakaryocytic cell surface markers in response to 12-O tetradecanoylphorbol-13-acetate (TPA). We conclude that downregulation of WT1 during induced differentiation of K562 cells is not a prerequisite for erythroid or megakaryocytic differentiation of these cells. PMID- 10378896 TI - Signal transduction pathways in normal human monocytes stimulated by cytokines and mediators: comparative study with normal human neutrophils or transformed cells and the putative roles in functionality and cell biology. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL) -3 induced tyrosine phosphorylation of 92-kDa protein in normal human monocytes. We identified this 92-kDa protein as STAT5, but not as STATs1, 3, and 6 nor c-fes and vav protooncogene products, and demonstrated its translocation to the nucleus, enhancement of specific DNA binding capacity, and potentiation of trancriptional activity by GM-CSF. N-formyl-methionyl-leucyl-phenylalanine (FMLP) and phorbol myristate acetate (PMA) induced tyrosine phosphorylation of 42- and 44-kDa proteins, which were identified as extracellular signal-regulated kinase (ERK), in human monocytes. In marked contrast to neutrophils and MO7e cells, GM CSF did not induce tyrosine phosphorylation and activation of ERK in monocytes. Among upstream signaling molecules of ERK, Shc was constitutively associated with Grb2 and was not tyrosine-phosphorylated by GM-CSF and FMLP, and Sos1 and c-Raf-1 were not phosphorylated by GM-CSF, IL-3, TNF, and FMLP in monocytes, whereas all these signaling molecules were affected and/or utilized by GM-CSF in MO7e cells. In contrast to neutrophils, p38 was constitutively phosphorylated and agonist dependent phosphorylation and activation was not detected in human monocytes. Superoxide release stimulated by FMLP was inhibited partially by PD98059 or SB203580, a specific inhibitor of ERK or p38 pathway, and was almost completely inhibited by the combination of both inhibitors, whereas PMA-induced superoxide release was resistant to these two inhibitors in monocytes. PD98059 inhibited GM CSF-dependent proliferation of MO7e cells. Present results indicate trancriptional roles of STAT5 and functional roles of ERK and/or p38 in normal human monocytes stimulated by physiological receptor-mediated agonists GM-CSF and FMLP. Possible roles of ERK in proliferation of transformed cells were also suggested. PMID- 10378897 TI - Changes in signal transduction downstream from the granulocyte-macrophage colony stimulating factor receptor during differentiation of primary hemopoietic cells. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a multifunctional cytokine, having different effects on primitive hemopoietic cells and terminally differentiated end-cells of the myeloid lineage. Human primitive hemopoietic cells (CD34+) were obtained from the peripheral blood after mobilization and induced to proliferate and then differentiate with a combination of cytokines in vitro. Cells at different time points were then used to analyze the expression of the GM-CSF receptor and GM-CSF mediated activation of the JAK 2-STAT 5 and MAP kinase pathways. Scatchard analysis as measured by radioligand binding revealed that freshly purified CD34+ cells expressed 36+/-1 high affinity receptors per cell (mean +/- SE, n = 3) and the level of expression was not significantly different after 3 days in culture, but rose five- to tenfold by day 8. The day 0 CD34+ cells were hyporesponsive to GM-CSF, but by 3 days in culture the cells were still morphologically immature but were actively proliferating and exhibited maximal GM-CSF induced JAK 2-STAT 5 and MAP kinase activation at the optimal time point. Further culture of the CD34+ cells resulted in myeloid differentiation associated with prolongation of MAP kinase activation but not JAK 2-STAT 5 activation. These data indicate that the JAK 2-STAT 5 and MAP kinase pathways are independently regulated and that changes in these signaling pathways occur with differentiation. PMID- 10378898 TI - A novel stromal cell-dependent hematopoietic cell line established from temperature-sensitive SV40 T-antigen transgenic mice. AB - A novel primitive hematopoietic cell line, THS119, was established from lineage marker negative (Lin-)/Sca-1+ cells from bone marrow of temperature-sensitive (ts) SV40 T-antigen transgenic mice after lengthy passaging by coculture with TBR59 bone marrow stromal cells. THS119 cells exhibited immature primitive hematopoietic cells such as forming cobblestones underneath the stromal cell layers. They retained properties of hematopoietic stem cells as shown by expression of c-Kit, Sca-1 and CD34low, but lacked hematopoietic lineage surface markers of differentiated hematopoietic cells (Gr-1, TER119, Mac-1, CD3, B220). RT-PCR analysis showed that THS119 cells exhibited multiple expression of both earlier developmental markers of myeloid, lymphoid and the hematopoietic cell specific transcription factors. THS119 cells showed temperature-dependent growth reflecting ts T-antigen, and their maintenance was TBR59 stromal cell-dependent. The requirement of stromal cells could not be replaced by cytokines, however, an IL-3 or IL-7 dependent cell line was generated after prolonged culture of THS119 cells on the stromal cells in the presence of these cytokines, and these cytokine dependent cell lines exhibited phenotypes similar to the parental cells in their gene expression. SCF/c-Kit interaction is one factor required for their maintenance, but involvement of other factor(s) in the conditioned medium of TBR59 stromal cells was suggested. A novel immature hematopoietic cell line, THS119, may provide an appropriate experimental system to resolve how hematopoietic cells are kept in a primitive phase within a hematopoietic microenvironment. PMID- 10378899 TI - Characterization and retroviral transduction of an early human lymphomyeloid precursor assayed in nonswitched long-term culture on murine stroma. AB - In the hierarchy of human hematopoietic progenitors, long-term culture-initiating cells (LTC-IC) and extended LTC-IC belong to the earliest cell populations that can be assayed in vitro. We report the identification of a multipotential lymphomyeloid progenitor detected in a nonswitch culture system. We observed the emergence of CD33+ myeloid and CD19+ B-lymphoid cells following plating of lineage-depleted (Lin-) CD34 -enriched or purified CD34+ CD38- cord blood cells on MS-5 stroma in the absence of exogenous cytokines. Both CD19+ CD20- pro-B and CD19+ CD20+ pre-B lymphocytes coexist with myeloid cells in long-term culture. A limiting dilution approach was used to show that a single CD34+ CD38- cell can generate lymphomyeloid progeny in conventional (5-week) and extended (10-week) cultures. Most of the clones in long-term culture or extended long-term culture contained not only lymphoid and myeloid cells, but also myeloid clonogenic progenitors. A high proportion of CD34+ CD38- cells gave rise to lymphomyeloid clones after 5 and 10 weeks of culturing (up to 48% and 16%, respectively), which distinguishes the assay reported here from those using switch culture conditions. We performed retroviral gene transfer experiments involving 1-3 days of exposure of Lin CD34+ -enriched cells to virus encoding enhanced green fluorescent protein. Monitoring of gene transfer efficiency into LTC-IC by enhanced green fluorescent protein fluorescence showed that it is possible to achieve marking of lymphomyeloid LTC-IC, albeit to a lesser extent than myeloid-restricted LTC-IC. PMID- 10378900 TI - Exercise performance in those having Parkinson's disease and healthy normals. AB - OBJECTIVE: This study assessed and compared the cardiopulmonary function of individuals with Parkinson's disease (PD) with that of healthy normals (HN) in order to provide health professionals with more thorough information about the problems associated with PD. METHODS: 20 men (PD = 13, HN = 7; mean age 64 and 64, respectively) and 23 women (PD = 7, HN = 16; mean age 65 and 66, respectively) were recruited from the Houston metropolitan area. Maximal oxygen consumption (VO2max mL x kg(-1) x min(-1)) and time to maximal exercise in minutes (time(max)) were measured. Exercise was performed on a stationary bicycle using an incremental exercise protocol. Because the assumption of homogeneity of variance was not met for the dependent variable VO2max in women, the nonparametric Wilcoxon-Mann-Whitney-U analysis was used (alpha < or = 0.025). All other group comparisons were analyzed using an independent t-test (alpha < or = 0.025). RESULTS: For men and women, there were no significant differences in VO2max between those having PD and the HN (men: PD = 23.52 vs HN = 25.46 mL x kg( 1) x min(-1), P = 0.50; women: PD = 20.10 vs HN = 16.20 mL x kg(-1) x min(-1), P = 0.35). Likewise, there was no significant differences in time(max) between women (PD = 5.2 vs HN = 5.4 min, P = 0.20). Comparison of time(max) between men did show a significant difference (PD = 9.5 vs HN = 13.10 min, P = 0.02). CONCLUSIONS: Although there were no significant differences in VO2max between the men, the comparison of time(max) indicates those with PD were unable to exercise as long before reaching VO2max, indicating that individuals with PD may be less efficient during exercise and therefore unable to exercise as long before reaching VO2max. Although women with PD had a higher VO2max, comparisons of VO2max and time(max) between those with PD and HN resulted in no significant differences. PMID- 10378901 TI - Gastrointestinal symptoms during long-distance walking. AB - PURPOSE: Gastrointestinal (GI) symptoms are common during prolonged intense exercise. To examine whether GI symptoms are also common during prolonged exercise of lower intensity, we obtained data on incidence, duration, and severity of GI symptoms during four consecutive days walking with a total distance of 203 km for men and 164 km for women. METHODS: The research population consisted of 79 men and 76 women, aged 30-49 yr, who responded to a questionnaire and a diary concerning anthropometric data, activity pattern, dietary intake, and GI symptoms. RESULTS: The results show that 24% of the subjects experienced one or more symptoms. Nausea, headache, and flatulence were the most frequent symptoms. Nine subjects dropped out during the race, two of whom indicated that they stopped as a result of one or more GI symptoms. Logistic regression analysis revealed that the occurrence of GI symptoms was a significant exercise-limiting factor. Univariate analysis showed that incidence and duration of GI symptoms were significantly related to the subjects' experience (number of prior participations to the event), body weight loss during walking, and several components of the diet before and during the event. A significant relationship between GI symptoms and age, gender, training status, and walking speed could not be found. CONCLUSIONS: We conclude that GI symptoms during long-distance walking can impair exercise performance, although these symptoms occur less frequently and are less severe in comparison with prolonged intense exercise. PMID- 10378902 TI - Preventing dehydration in children with cystic fibrosis who exercise in the heat. AB - PURPOSE: In healthy children who exercise in the heat, the addition of flavor, carbohydrate, and 18 mmol x L(-1) NaCl to water induced a major increase in voluntary drink intake compared with the intake of unflavored water. This increase was sufficient to prevent voluntary dehydration. We hypothesized that, to achieve a similar effect in children with cystic fibrosis (CF), whose NaCl losses in sweat are markedly excessive, the drink should include an NaCl concentration higher than 18 mmol x L(-1). METHODS: Eleven subjects with CF (6 girls, 5 boys, ages 10.9-19.5 yr) attended three 3-h sessions of intermittent exercise of moderate intensity (four 20-min bouts), at 35 degrees C, 50% relative humidity. Either water (W), flavored water (FW), or a 30 mmol x L(-1) NaCl plus 6% carbohydrate solution (Na30) was offered ad libitum, in a counterbalanced sequence. Six subjects performed an additional session in which they drank a 50 mmol x L(-1) NaCl-6% CHO solution (Na50). RESULTS: There was no significant drink effect on body fluid balance, core temperature, heart rate, or serum electrolytes with W, FW, or Na30. Serum osmolality decreased throughout the sessions from 290.6 +/- 1.1 (mean +/- SEM) to 281.3 +/- 1.2 mmol x kg(-1) (P < 0.0005), serum sodium from 143.1 +/- 0.5 to 141.1 +/- 0.7 mmol x L(-1) (P = 0.01) and serum chloride from 109.1 +/- 0.5 to 107.5 +/- 0.5 mmol X L(-1) (P < 0.001). In contrast, the 50 mmol x L(-1) NaCl drink induced a near significant (P = 0.08) higher fluid intake, and it significantly ameliorated the rate of progressive dehydration. CONCLUSIONS: The marked loss of NaCl in the sweat of CF patients may induce an hypo-osmolar state in the serum, even when the drink contains 30 mmol x L(-1) NaCl. This may diminish the thirst drive triggered by hypothalamic osmoreceptors and may lead to voluntary dehydration. A flavored drink with an even higher salt content (50 mmol X L(-1)), however, enhances drinking and attenuates the voluntary dehydration. PMID- 10378904 TI - Development and clinical application of kinematic MRI of the patellofemoral joint using an extremity MR system. AB - PURPOSE: Kinematic magnetic resonance imaging (MRI) of the patellofemoral joint provides diagnostic information pertaining to patellar alignment and tracking during the earliest increments of joint flexion, when abnormalities that affect this joint are the most apparent. Recently, a low-field strength (0.2 Tesla) dedicated extremity MR system has been designed, such that only the body part that is being imaged is placed inside of the magnet bore. The purpose of this investigation was to develop a kinematic MRI technique for the patellofemoral joint using the extremity MR system and to apply this procedure in the clinical setting. METHODS: An incremental, passive positioning kinematic MRI technique was developed for the patellofemoral joint that involved obtaining three different axial section locations with the patellofemoral joint extended and then imaging these same section locations repeatedly as the patellofemoral joint was flexed in four increments up to 36 degrees of flexion. MR images were obtained using a T1 weighted spin echo sequence. Five (10 PFJ) asymptomatic volunteers and nine patients (9 PFJ) with patellofemoral joint symptoms were studied. RESULTS: Volunteers had normal kinematic MRI examinations. Seven patients had lateral subluxation, and two patients had excessive lateral pressure syndrome. Two patients with lateral subluxation seen on their kinematic MRI studies had Merchant views (x-rays obtained at 45 degrees) that showed "normal" patellar alignment, illustrating the importance of imaging the patellofemoral joint at 30 degrees or less. CONCLUSIONS: A kinematic MRI technique was successfully developed for the low-field extremity MR system and utilized for clinical applications. This procedure may be used to determine the presence and severity of patellar malalignment and abnormal tracking patterns. PMID- 10378903 TI - Running and ovulation positively change cancellous bone in premenopausal women. AB - PURPOSE: Exercise is understood to exert positive effects on bone. However cancellous bone has not been shown to increase with exercise. Previous results of our 1-yr observational prospective study in ovulatory women related 20% of the change in cancellous spinal bone mineral density (BMD), measured by quantitative computed tomography (QCT), to luteal phase length (the time from ovulation to menstruation, LL). METHODS: The 66 women who documented exercise daily included normally active women (N = 23) and those who ran consistently or were increasing running in preparation for a marathon (N = 43). Exercise did not affect BMD change in the women as a whole. We re-evaluated those data to determine whether exercise-related effects on spinal cancellous BMD change in regularly cycling premenopausal women were related to ovulatory characteristics. The potential relationship of exercise to BMD change was reanalyzed by stratifying women into tertiles according to average LL documented by quantitative basal temperature analysis. RESULTS: Repeated-measures ANOVA indicated independent positive effects of both luteal length (P = 0.001) and activity (P = 0.041). The 11 runners with LL > 10.9 d had a nonsignificant 0.5% increase in lumbar BMD while the 15 who averaged short LL (<9.9 d) experienced a significant 3.6% loss. In the runners as a group, however, kilometers run per week was negatively related to BMD change throughout (r = -0.347, P = 0.024). CONCLUSIONS: These data are the first to indicate that, in women with regular cycles, luteal length and exercise independently and positively affect change in spinal cancellous BMD. PMID- 10378905 TI - Physical conditioning effects on fetal heart rate responses to graded maternal exercise. AB - PURPOSE: This study examined the effects of advancing gestational age and maternal aerobic conditioning (stationary cycling) on fetal heart rate (FHR) responses to strenuous non-steady-state maternal exercise. METHODS: Subjects chose to participate in either an exercise group (EG) or control group (CG). Fourteen healthy, previously sedentary pregnant women participated in the exercise group, and six pregnant controls remained sedentary. Stationary cycling (heart rate target: 145 beats x min(-1)) was performed 3 d x wk(-1) by the exercised group. Exercise duration was increased from 14 to 25 min x session(-1) during the second trimester and was maintained at 25 min x session(-1) throughout the third trimester. FHR was monitored before, during, and after a progressive submaximal cycle ergometer test (peak heart rate = 170 beats x min(-1)) performed at approximately 27 and 37 wk gestation. RESULTS: Mean FHR increased significantly (P < 0.05) during exercise, followed by a modest suppression and then a delayed rise during the recovery period at both observation times. Fetal bradycardia was not observed in any of the exercise tests. Effects of advancing gestational age included a lower FHR baseline both at rest and in response to maternal exercise and a lower incidence of exercise-induced tachycardia. Maternal physical conditioning did not significantly alter FHR response to maternal exercise. CONCLUSION: Our results support the hypothesis that FHR responses to strenuous exercise are altered by advancing gestational age and a brief progressive exercise test terminated at a maternal heart rate of 170 beats x min( 1) does not induce fetal distress during a healthy pregnancy. PMID- 10378906 TI - Induced hypervolemia, cardiac function, VO2max, and performance of elite cyclists. AB - OBJECTIVE: To determine whether plasma volume expansion (PVexp) in elite endurance-trained (ET) cyclists, who already possess both a high blood volume (BV) and a high VO2max, leads to further enhancements in their cardiac function, VO2max, and endurance performance (time to exhaustion at 95% VO2max). METHODS: Nine male ET cyclists (V02max = 68.9 +/- 0.6 (SEM) mL x kg(-1) x min(-1)) were studied employing a double blind, cross-over design; i) before PVexp, ii) after sham PVexp (Sham), iii) after restoration of normocythemia, iv) after PVexp (6% dextran), and v) upon reestablishment of normocythemia. RESULTS: PVexp resulted in a 547 +/- 61 mL increase in BV (P < 0.05). Maximal cardiac output and maximal stroke volume were higher (P < 0.05) after PVexp, but the magnitude of these increases was only sufficient to counter the hemodilution effect (lowered O2 content) of PVexp, such that O2 transport, VO2max, and endurance performance remained unchanged. CONCLUSIONS: Expansion of BV in elite ET cyclists, who already possess a high BV, does not improve their VO2max and endurance performance. Elite ET athletes may already be at an optimal BV, which is at or near the limits of their diastolic reserve capacity. PMID- 10378907 TI - Hyponatremia in ultradistance triathletes. AB - PURPOSE: Hyponatremia ([plasma sodium] <135 mmol x L(-1)) is a potentially serious complication of ultraendurance sports. However, the etiology of this condition is still uncertain. This observational cohort study aimed to determine prospectively the incidence and etiology of hyponatremia in an ultradistance triathlon. METHODS: The subjects consisted of 605 of the 660 athletes entered in the New Zealand Ironman triathlon (3.8-km swim, 180-km cycle, and 42.2-km run). Subjects were weighed before and after the race. A blood sample was drawn for measurement of plasma sodium concentration after the race. RESULTS: Complete data on pre- and postrace weights and plasma sodium concentrations were available in 330 race finishers. Postrace plasma sodium concentrations were inversely related to changes in body weight (P = 0.0001). Women (N = 38) had significantly lower plasma sodium concentrations (133.7 vs 137.4 mmol x L(-1); P = 0.0001) than men (N = 292) and lost significantly less relative weight (-2.7 vs -4.3%; P = 0.0002). Fifty-eight of 330 race finishers (18%) were hyponatremic; of these only 18 (31%) sought medical care for the symptoms of hyponatremia (symptomatic). Eleven of the 58 hyponatremic athletes had severe hyponatremia ([plasma sodium] < 130 mmol x L(-1)); seven of these 11 severely hyponatremic athletes were symptomatic. The relative body weight change of the 11 severely hyponatremic athletes ranged from 2.4% to +5%; eight (73%) of these athletes either maintained or gained weight during the race. In contrast, relative body weight changes in the 47 athletes with mild hyponatremia ([plasma sodium] 130-134 mmol x L(1)) were more variable, ranging from -9.25% to +2.2%. CONCLUSIONS: Hyponatremia is a common biochemical finding in ultradistance triathletes but is usually asymptomatic. Although mild hyponatremia was associated with variable body weight changes, fluid overload was the cause of most (73%) cases of severe, symptomatic hyponatremia. PMID- 10378909 TI - Cardiorespiratory responses to arm cranking and electrical stimulation leg cycling in people with paraplegia. AB - PURPOSE: The purpose of this study was to assess the cardiorespiratory responses during arm exercise with and without concurrent electrical stimulation-induced leg cycling in people with paraplegia. METHODS: On separate days, 10 subjects with spinal cord injuries (T5-T12) performed either arm cranking (ACE), or simultaneous arm cranking + electrical stimulation-induced leg cycling (ACE+ES LCE) graded exercise tests. RESULTS: During submaximal, steady-state exercise, ACE+ES-LCE elicited significantly higher VO2, (by 0.25-0.28 L x min(-1)) stroke volume (by 13 mL), and VE(BTPS) (by 9.4 L x min(-1)) compared with ACE alone. In contrast, there were no significant differences of submaximal HR, cardiac output, or power output between the exercise modes. At maximal exercise, ACE+ES-LCE elicited significantly higher VO2 (by 0.23 L x min(-1)) compared with ACE alone, but there were no differences in power output, HR, or VE(BTPS). CONCLUSIONS: These results demonstrate that during submaximal or maximal exercise there was a greater metabolic stress elicited during ACE+ES-LCE compared with during ACE alone. The higher stroke volume observed during submaximal ACE+ES-LCE, in the absence of any difference in HR, implied a reduced venous pooling and higher cardiac volume loading during ACE+ES-LCE. These results suggest that training incorporating ACE+ES-LCE may be more effective in improving aerobic fitness in people with paraplegia than ACE alone. PMID- 10378908 TI - Exercise training and heart rate variability in older people. AB - PURPOSE: Heart rate variability (HRV), a characteristic that is potentially increased by physical activity, has been associated with incidence of cardiac events and total mortality. Since the incidence of cardiac events among older people is high and their physical activity levels and HRV are generally low, it is important to investigate whether regular physical activity can modify HRV in this age group. The purpose of the study was to investigate the effect of regular physical activity on HRV in older men and women. METHODS: In a randomized controlled trial, the effect of six months' training on HRV was investigated in a group of 51 older men and women (67.0 +/- 5.1 yr). The training group gathered three times per week for 45 min supervised training. RESULTS: At the end of the intervention period, HRV was higher primarily during the day. During daytime, the SD of all normal intervals (+6%) as well as the low frequency component (+ 15%) and the very low frequency component (+ 10%) of HRV were significantly increased (P < 0.05) as compared with the control group. Effects of training were most pronounced in subjects inactive in sports at baseline. CONCLUSION: This study demonstrates that regular physical activity increases HRV (specifically in the very low and low frequency components) in older subjects. Hence, in older subjects, physical training may be an effective means to modify positively a factor that is associated with increased incidence of cardiac events. PMID- 10378910 TI - Differential leukocytosis and lymphocyte mitogenic response to acute maximal exercise in the young and old. AB - Despite the increasing use of exercise in the elderly as a means of improving muscle function, little is known regarding the effects of exercise on the senescent immune system. PURPOSE: The purpose of this study was to determine the effects of acute maximal exercise on blood leukocyte numbers, leukocyte subsets, and the T cell mitogenic response in the elderly. METHODS: Previously sedentary elderly (N = 33, 65.3 +/- 0.8 yr) and young (N = 14, 22.4 +/- 0.7 yr) subjects participated in a modified Balke maximal exercise treadmill test. Venous blood samples were collected pre-, immediately post-, and 20 min postexercise. Blood was analyzed for leukocyte counts, leukocyte subsets via immunofluorescence, and whole blood mitogenesis in response to various doses of mitogens. RESULTS: Whereas VO2max was lower in the elderly, maximal RQ, age-predicted heart rates, and times to fatigue were not different, indicating that both groups achieved relative maximal exercise intensity. There were significant exercise-induced leukocytoses in both the elderly and young made up largely of a lymphocytosis and neutrophilia. The magnitude of the leukocytosis was lower in the elderly and failed to return to pre-exercise levels by 20 min postexercise. Acute maximal exercise increased CD8+ (153% vs 112% in young and old, respectively) and CD4+ (57% vs 22% in young and old, respectively) T cells when measured immediately postexercise. By 20 min postexercise, concentrations in the young were not significantly different from baseline, whereas CD8 cell number was still elevated in the old. The elderly had significantly higher percentages of memory (i.e., CD45RO+) and significantly lower percentages of naive (i.e., CD45RA+) CD4 and CD8 T cells pre-exercise, and the young and old recruited approximately equal numbers of CD8+ naive and memory cells to the blood in response to exercise. In contrast, the aged recruited significantly fewer numbers of CD4+ naive and transitional (CD45RA+RO+) cells. At most doses of Con A and PHA, the lymphoproliferative response was lower in the elderly subjects even though they had significantly higher numbers and percentages of CD3+ cells. Interestingly, immediately postexercise, young (but not old) subjects demonstrated reduced proliferative ability on a per CD3+ cell basis. CONCLUSIONS: These data indicate that several blood leukocyte responses to maximal exercise stress are similar in the young and the old. However, the elderly demonstrate a less resilient leukocytosis and a different lympho-proliferative response following acute maximal exercise. PMID- 10378911 TI - Effect of incremental test protocol on the lactate minimum speed. AB - PURPOSE: The purpose of this study was to investigate the effect of altering the initial running speed (RS) in the incremental portion of the lactate minimum test on the lactate minimum speed (LMS). METHODS: Eight well-trained endurance runners (mean +/- SD age 29.0 +/- 5.4 yr, body mass 72.0 +/- 5.6 kg, VO2max 63.1 +/- 3.8 mL x kg(-1) min(-1)) completed a standard incremental treadmill test for the assessment of the lactate threshold (LT) and VO2max, and eight lactate minimum tests. Following a period of supramaximal exercise, subjects were allowed 8 min of recovery to allow blood [lactate] to peak. Subjects then undertook eight randomly-assigned incremental treadmill tests from different initial running speeds (3.0, 2.5, 2.0, 1.5, 1.0, and 0.5 km x h(-1) below the predetermined RS LT, at the RS-LT, and at 1.0 km x h(-1) above the RS-LT) with RS increased by 1.0 km x h(-1) every 5 min until volitional fatigue. Blood samples for the determination of blood [lactate] were taken at the end of each stage and the LMS was determined by fitting a spline function to the data. RESULTS: No LMS could be determined for the two highest initial RS conditions. For the other conditions, the LMS was significantly affected by the initial RS used in the incremental test and varied from 13.8 +/- 0.7 km x h(-1) with an initial RS of 3.0 km x h(-1) below the RS-LT, to 15.8 +/- 0.8 km x h(-1) with an initial RS of 0.5 km x h(-1) below the RS-LT. The LMS was significantly different from the RS-LT (15.4 +/- 0.8 km x h(-1)) (P < 0.05), except when the incremental test started at 1.0 or 1.5 km x h(-1) below the RS-LT. CONCLUSIONS: These results suggest that the LMS test is not a valid method for estimation of the LT since it is profoundly influenced by the starting speed selected for the incremental portion of the test. PMID- 10378912 TI - Muscle activation during the tennis volley. AB - PURPOSE: To broaden our understanding of muscle function during the tennis volley under different ball placement and speed conditions by examining the activity of selected superficial muscles of the stroking arm and shoulder (flexor carpi radialis, extensor carpi radialis, triceps brachii, deltoids, and pectoralis major) and muscles related to postural support (left and right external oblique, lumbar erector spinae, and gastrocnemius) during the volley. METHODS: Seven skilled tennis players were asked to perform volley strokes under 18 experimental conditions, including variations in lateral contact location (forehand and backhand), ball contact height (high, middle, and low), and ball speed (fast, medium, and slow). A ball machine was modified so that the subjects could not predict the ball trajectory before it was released from the machine. Muscle activity was determined using surface electromyographic (EMG) techniques, and the critical instants of a volley were determined using two force platforms and two high-speed (120 Hz) video cameras. Average EMG values for different phases of the volley, defined by the critical instants, were computed. RESULTS AND CONCLUSIONS: In general, muscle activity increased with increasing ball speed. The extensor carpi radialis was more active than the flexor carpi radialis during both forehand and backhand volleys, suggesting the importance of wrist extension/abduction and grip strength. The increase in EMG levels in the forearm muscles shortly before the ball impact indicated that the subjects did not tighten their grip and wrist until moments before ball impact. Both antero-middle and postero-middle deltoids were active in most stroke phases. However, the roles of the deltoid muscles during a volley cannot be determined without knowing the actions of the other shoulder joint muscles. PMID- 10378913 TI - Movement characteristics of the tennis volley. AB - PURPOSE: The purpose of this study was to examine selected movement characteristics of the tennis volley by evaluating temporal and ground reaction force (GRF) parameters. METHODS: Seven skilled tennis players performed volley strokes under 18 experimental conditions including variations in lateral contact location (forehand (FH) and backhand (BH)), ball contact height (high, middle, low), and ball speed (fast, medium, slow). A ball machine was modified so that the subjects could not predict the ball trajectory before it was released from the machine. The GRF and temporal parameters were determined using two force platforms and two high-speed (120 Hz) video cameras, respectively. Average and maximum values of each measured parameter were computed for different phases of the volley. RESULTS: The average reaction times (from ball release to initial racquet movement (IRM)) for FH and BH trials were 226 and 205 ms, respectively, and the difference was statistically significant. The average stroke time (from IRM to ball impact) ranged from 381 ms in fast speed trials to 803 ms in slow speed trials. A distinct racquet forward motion immediately before ball impact occurred in 75% of the trials and they were evenly distributed between FH and BH trials. An ipsilateral side step (a side step of the foot on the same side of the oncoming ball before the crossover step of the other foot) occurred more often in FH (45%) than in BH (34%) trials. CONCLUSIONS: The GRF during the stroke phase suggest that the subjects initiated lateral movement by leaning sideward when ball velocity was low and by a vigorous pushoff of the contralateral foot when ball velocity was high. PMID- 10378914 TI - Is sleep disturbed by vigorous late-night exercise? AB - PURPOSE: This experiment examined the influence of prolonged, vigorous late-night exercise on sleep. METHODS: Sixteen highly fit male cyclists completed each of two 60-h laboratory treatments involving a baseline night, an experimental treatment night, and a recovery night. In counterbalanced order, subjects 1) cycled for 3 h at 65-75% of heart rate reserve combined with bright light exposure (3000 lux) light, and 2) were exposed to a 3 h pulse of bright light (3000 lux) alone. RESULTS: On the baseline and recovery nights, subjects maintained their usual sleep-wake schedules. On the treatment night, exercise + bright light or bright light alone were centered at 6 h before their usual wake times, followed by bedtimes 30 min after the treatments. Illumination was 3000 lux during the experimental treatments, 0 lux during the sleep periods, and 50 lux at other times. Sleep was assessed with an Actillume (Ambulatory Monitoring, Inc., Ardsley, NY) wrist monitor to define sleep onset latency (SOL), wakefulness after sleep onset (WASO), and total sleep time. Subjective assessments of SOL, WASO, and insomnia were also gathered each morning. No significant differences in objective or subjective sleep variables were found between treatments. CONCLUSIONS: These data are inconsistent with the general opinion that vigorous exercise shortly before bedtime disturbs sleep. PMID- 10378915 TI - Maximal strength training improves work economy in trained female cross-country skiers. AB - PURPOSE: The present study examines the hypothesis that maximal strength training improves work economy and anaerobic threshold in trained female cross-country skiers while working on a ski ergometer. METHODS: Fifteen female cross-country skiers (17.9 +/- 0.3 yr, 166.7 +/- 1.3 cm, 60.1 +/- 1.9 kg, and 55.3 +/- 1.3 mL x kg(-1) x min(-1)) participated in the study. Eight skiers made up the high intensity strength-trained group, and seven served as the control group. Endurance performance was tested on a specially instrumented ski ergometer. Strength training and testing simulated double poling in cross-country skiing. RESULTS: A significant (P < 0.001) improvement in double-poling economy on the ski ergometer was observed among the strength-trained group. Anaerobic threshold did not change during the experimental period for either group. After a 9-wk training period, time to exhaustion increased from 5.2 (+/-0.9) to 12.3 (+/-1.6) min (P < 0.001) and from 4.0 (+/-0.9) to 6.3 (+/-0.9) min (P < 0.01) for the strength and control group, respectively. Time to exhaustion was significantly higher (P < 0.001) for the strength group compared with the control group after training. One repetition maximum increased 14.5% (1.8) (P < 0.001) in the strength group but was unchanged in the control group. Expressed in relation to peak force at one repetition maximum, strength training resulted in a significant reduction in the relative available force employed working on the ski ergometer (P < 0.01). Time to peak force at maximal aerobic velocity on the ski ergometer was significantly reduced in the strength-training group (P < 0.01). CONCLUSIONS: It is concluded that maximal strength training in the upper-body improved the double-poling performance by improved work economy. Work economy was improved by a reduction in relative workload and time to peak force while double poling. PMID- 10378916 TI - Level ground and uphill cycling ability in professional road cycling. AB - PURPOSE: To evaluate the physiological capacities and performance of professional road cyclists in relation to their morphotype-dependent speciality. METHODS: 24 world-class cyclists, classified as flat terrain (FT, N = 5), time trial (TT, N = 4), all terrain (AT, N = 6). and uphill (UH, N = 9) specialists, completed an incremental laboratory cycling test to assess maximal power output (Wmax), maximal oxygen uptake (VO2max), lactate threshold (LT), and onset of blood lactate accumulation (OBLA). RESULTS: UH had a higher frontal area (FA):body mass (BM) ratio (5.23 +/- 0.09 m2 x kg(-1) x 10(-3)) than FT and TT (P < 0.05). FT showed the highest absolute Wmax (481 +/- 18 W), and UH the highest Wmax relative to BM (6.47 +/- 0.33 W x kg(-1)). WLT and W(OBLA) values were significantly higher in FT (356 +/- 41 and 417 +/- 45 W) and TT (357 +/- 41 and 409 +/- 46 W) than in UH (308 +/- 46 and 356 +/- 41). Scaling of these values relative to FA and BM exponents 0.32 and 0.79 minimized group differences, but considerable differences among mean group values remained. FT and TT had the highest Wmax per FA unit (1300 +/- 62 and 1293 +/- 57 W x m2), whereas TT had the highest absolute W x kg(-0.32) and W x kg(-0.79), as well as W x kg(-0.32), W x kg(-0.79), and W x m2 at the LT and OBLA. CONCLUSIONS: i) Scaling of maximal and submaximal physiological values showed a performance advantage of TT over FT, AT, and UH in all cycling terrains and conditions; and ii) mass exponents of 0.32 and 1 were the most appropriate to evaluate level and uphill cycling ability, respectively, whereas absolute Wmax values are recommended for performance-prediction in short events on level terrain, and W(LT) and W(OBLA) in longer time trials and uphill cycling. PMID- 10378918 TI - Effects of 4-wk training using Vmax/Tmax on VO2max and performance in athletes. AB - PURPOSE: The aim of this study was to determine the effects of a 4-wk individualized training program using Vmax as the exercise intensity and utilizing between 60 and 75% of a subject's Tmax as the exercise duration. METHODS: Five male, middle-distance, trained subjects with the following characteristics (mean +/- SD): age, 22.8 +/- 4.5 yr; height, 181 +/- 4.7 cm; weight, 74.1 +/- 3.2 kg; skinfolds based on five areas, 35.9 +/- 3.9; and VO2max, 61.5 +/- 6.1 mL O2 x kg min(-1) volunteered to participate in this study. Before the training program, the subjects completed a 3000-m time trial, and three each of VO2max/Vmax and Tmax tests. Subjects then completed a 4-wk training program on the treadmill and were then retested on the VO2max/Vmax and Tmax tests. RESULTS: Pretraining versus posttraining results showed significant (P < 0.05) increases in average Vmax (20.5 km x h(-1) vs 21.3 km x h(-1) posttraining), Tmax (225.5 s vs 300.9 s posttraining), and VO2max (61.5 mL O2 x kg x min(-1) vs 64.5 mL O2 x kg x min(-1)). The 3000-m time trial decreased significantly from a pretraining value of 616.6 s to a posttraining value of 599.6 s (P < 0.05). CONCLUSIONS: The results of this study indicate that by utilizing between 60 and 75% of Tmax as an exercise duration and using Vmax as an exercise intensity that these two parameters can be extremely valuable in the prescription of exercise programs for athletes. PMID- 10378917 TI - The effects of strength training on endurance performance and muscle characteristics. AB - PURPOSE: The purpose of this study was to determine the effects of resistance training on endurance performance and selected muscle characteristics of female cyclists. METHODS: Twenty-one endurance-trained, female cyclists, aged 18-42 yr, were randomly assigned to either a resistance training (RT; N = 14) or a control group (CON; N = 7). Resistance training (2X x wk(-1)) consisted of five sets to failure (2-8 RM) of parallel squats for 12 wk. Before and immediately after the resistance-training period, all subjects completed an incremental cycle test to allow determination of both their lactate threshold (LT) and peak oxygen consumption VO2). In addition, endurance performance was assessed by average power output during a 1-h cycle test (OHT), and leg strength was measured by recording the subject's one repetition maximum (1 RM) concentric squat. Before and after the 12-wk training program, resting muscle was sampled by needle biopsy from m. vastus lateralis and analyzed for fiber type diameter, fiber type percentage, and the activities of 2-oxoglutarate dehydrogenase and phosphofructokinase. RESULTS: After the resistance training program, there was a significant increase in 1 RM concentric squat strength for RT (35.9%) but not for CON (3.7%) (P < 0.05). However, there were no significant changes in OHT performance, LT, VO2, muscle fiber characteristics, or enzyme activities in either group (P > 0.05). CONCLUSION: The present data suggest that increased leg strength does not improve cycle endurance performance in endurance-trained, female cyclists. PMID- 10378919 TI - Reliability and comparability of the accelerometer as a measure of muscular power. AB - PURPOSE: The purpose of this study was to determine the effectiveness of using accelerometry as a reliable measure of upper body muscular power and its comparability with other conventional measurement tools. METHODS: Thirty men, aged 19 to 25 yr, gave informed written consent before performing a one repetition maximum (1RM) bench press on a linear bench press apparatus. Three trials of 60% 1RM were then performed at 1-min intervals and the entire procedure was repeated the following day. Each trial was analyzed for average power (AP), average velocity (AV), and total displacement (TD) by three instruments: a uni axial piezoresistive accelerometer (ICS Sensors Model 3145, Milpitas, CA) mounted on the Cybex Smith Press (Owatonna, MN) apparatus, a 17-mm video camera that recorded the lift, and an infrared photocell and timer system arranged to analyze a 20-cm segment of the lift. Acceleration data collected at 60 Hz obtained a direct measurement of force and an integrated measure of velocity to calculate muscular power. RESULTS: Repeated measures ANOVA and intraclass correlation coefficients indicated high trial to trial reliability (r = 0.99) for all measurement variables. Film AP was significantly greater (P < or = 0.05) than the accelerometer AP and the photocell AP in the 20-cm segment (356.6 +/- 94.8 W vs 335.5 +/- 97.7 W, and 342.0 +/- 97.2 W, respectively). Also, significant mean differences (P < or = 0.05) between the accelerometer and film measurements existed for AP (246.0 +/- 70.2 W vs 286.1 +/- 83.6 W), AV (44.4 +/- 9.2 cm x s( 1) vs 51.3 +/- 12.3 cm x s(-1)) and TD (43.2 +/- 7.9 cm vs 47.4 +/- 7.4 cm) when examined over the entire lift, but there were significant correlations between the two methods (AP, r = 0.95; AV, r = 0.98; TD, r = 0.93). CONCLUSIONS: These results suggest that although minor data acquisition errors were present, accelerometers can provide a reliable and versatile means to assess muscle power. PMID- 10378920 TI - The heart rate turn point reliability and methodological aspects. AB - PURPOSE: The aim of the study was to test protocol variations on the heart rate performance curve (HRPC) and the heart rate turn point (HRTP) according to Conconi et al. (1996). Respiratory gas exchange variables were used to define three phases of energy supply (I, II, III). METHODS: Eighteen healthy young male subjects performed 4 tests (T1-T4). T1: initial speed of 6 km x h(-1) followed by increments of 0.6 km x h(-1) every 60 s. Subjects were than randomized for the next three tests. T2: initial speed 5.6 km x h(-1) followed by increments of 0.2 km x h(-1) every 20 s; T3: similar to T2, in the second half of phase III acceleration (S) was increased. T4: like T2, at the beginning of phase III, S was increased. No differences were found in the degree of the deflection of the HRPC expressed as factor kHR between T1 (0.228 +/- 0.225) and T2 (0.248 +/- 0.231) but a significant increase was found in T3 (0.533 +/- 0.248) and T4 (0.770 +/- 0.258). RESULTS: The modifications of the protocol (T3 and T4) systematically influenced the deflection of the HRPC, but kHR was highly reproducible in all tests. Eleven subjects showed degrees of deflection in the HRPC in all tests. There were no significant differences for S, HR, and VO2 at the HRTP. An HRTP was not found in seven subjects in neither T1 or T2; however, in T3 and T4, these seven subjects showed a deflection of HRPC resulting from the protocol. The HRTP was found to be dependent on the start of the acceleration in phase III. In cases with a linear time course in the HRPC in T1 and T2, in T3 an HRTP was found at 15.6 km x h(-1) and in T4 at 13.6 km x h(-1) , respectively. CONCLUSION: The Conconi test protocol with an accelerated increase in S in the final phase of the test has a major influence on the occurrence of the HRTP in cases of near linear HRPC. PMID- 10378921 TI - Validity, reliability, and calibration of the Tritrac accelerometer as a measure of physical activity. AB - PURPOSE: The purposes of this study were to assess the validity and reliability of the Tritrac R3D accelerometer during treadmill walking and running and then to calibrate the instrument. METHODS: The Tritrac was assessed on 60 young adults (23.4 +/- 2.9 yr) during treadmill walking and running at 3.2, 6.4, and 9.7 km x h(-1). The calibration was carried out by identifying ranges of Tritrac raw data (vector magnitude) values corresponding to light (2-3.9 MET), moderate (4-7 MET), and vigorous (>7 MET) physical activity. Energy expenditure (EE), measured by indirect calorimetry, served as the criterion measure. RESULTS: Interinstrument intraclass reliability coefficients for Tritracs worn on the right and left hip ranged from 0.73-0.87, while intersession coefficients demonstrated high reliability for all speeds (R = 0.87-0.92). Paired t-tests comparing mean accelerometer counts at 6.4 km x h(-1), 0% grade (2647 +/- 456), and 6.4 km x h( 1), 5% grade (2635 +/- 435) demonstrated no significant difference (P > 0.05). Mean differences between EE measured by indirect calorimetry and that estimated by the Tritrac ranged from 0.0082 kcal x kg(-1) x min(-1) at 3.2 km x h(-1) to 0.0320 kcal x kg(-1) x min(-1) at 9.7 km x h(-1), with the Tritrac consistently overestimating EE during horizontal treadmill walking. The relationship between vector magnitude and EE across all speeds was highly linear (R2 = 0.90, SEE = 0.014 kcal x kg(-1) x min(-1)), with little overlap between light, moderate, and vigorous categories. The mean vector magnitudes at 2, 4, and 7 MET were 650, 1772, and 3455, respectively. CONCLUSIONS: These data indicate that the Tritrac is highly reliable from day to day and is sensitive to changes in speed but not grade. Furthermore, the Tritrac accurately distinguishes various intensities of walking and jogging on level ground. With limitations, these cut-points can be used to categorize light, moderate, and vigorous physical activity and to estimate EE. PMID- 10378922 TI - Reliability of power output during intermittent high-intensity cycling. AB - PURPOSE: We determined the number of trials on consecutive days required to establish high reliability of an intermittent high-intensity cycling test in subjects unfamiliar with multiple-sprint exercise. We also examined the extent to which this reliability could be maintained for 6 d. METHODS: Five untrained men performed a multiple-sprint test (10 x 7 s, with each sprint separated by 30 s) on each of four consecutive days (days 1-4), then rested for 6 d, and finally performed two additional tests on consecutive days (days 11 and 12). For statistical comparisons (analyses of variance), mean power outputs during sprints 8, 9, and 10 (MP8-10) on each test day were calculated for each of the 4th, 5th, and 6th seconds of the sprints, i.e., MP8-10(4th), MP8-10(5th), and MP8-10(6th). Peak power during each sprint was also examined. RESULTS: For days 3 and 4, values for MP8-10(4th), MP8-10(5th), and MP8-10(6th) were greater than on day 1 (P < 0.05). MP8-10(6th) on day 2 was also greater than on day 1 (P < 0.05). There were no differences in MP8-10 among days 2, 3, 4, 11, and 12. Also, peak power on day 1 was lower (P < 0.05) than peak power for all other days, which were not different from one another. The coefficients of variation (CV) for MP8-10 on day 3 versus day 4 were 3.3%, 2.5%, and 2.9% for MP8-10(4th), MP8-10(5th), and MP8 10(6th), respectively. The CV for MP8-10(4th), MP8-10(5th), and MP8-10(6th) on days 4, 11, and 12 ranged from 2.1 to 3.9%, with an overall mean of 3.1%. The greatest CV for MP8-10 was 5.2% for MP8-10(6th) on days 2 versus 3 and 2 versus 4. The mean CV for peak power for all pairwise combinations of days 4, 11, and 12 was 2.8%. CONCLUSIONS: In conclusion, satisfactory reliability of intermittent cycling tests is achieved after two familiarization sessions identical to the tests, and that reliability can be maintained for 6 d. PMID- 10378923 TI - The quantity and quality of exercise for healthy adults. PMID- 10378924 TI - Permanent cardiac assistance from skeletal muscle: a prospect for the new millennium. AB - This paper looks at the prospects for new surgical solutions to the problem of end-stage heart failure based on cardiac assistance from skeletal muscle. The current status of the main biological approaches, cardiomyoplasty, aortomyoplasty, and the skeletal muscle ventricle, are discussed, followed by a consideration of some of the important basic issues that need to be addressed if these techniques are to achieve their full potential. Although there is a review element to the paper, the main emphasis is on the work of our own research group and collaborating workers. PMID- 10378925 TI - Three parameters optimizing closed-loop control in sequential segmental neuromuscular stimulation. AB - In conventional dynamic myoplasties, the force generation is poorly controlled. This causes unnecessary fatigue of the transposed/transplanted electrically stimulated muscles and causes damage to the involved tissues. We introduced sequential segmental neuromuscular stimulation (SSNS) to reduce muscle fatigue by allowing part of the muscle to rest periodically while the other parts work. Despite this improvement, we hypothesize that fatigue could be further reduced in some applications of dynamic myoplasty if the muscles were made to contract according to need. The first necessary step is to gain appropriate control over the contractile activity of the dynamic myoplasty. Therefore, closed-loop control was tested on a sequentially stimulated neosphincter to strive for the best possible control over the amount of generated pressure. A selection of parameters was validated for optimizing control. We concluded that the frequency of corrections, the threshold for corrections, and the transition time are meaningful parameters in the controlling algorithm of the closed-loop control in a sequentially stimulated myoplasty. PMID- 10378926 TI - New easy to install nerve cuff electrode using shape memory alloy armature. AB - This paper presents an easy to install nerve cuff electrode dedicated to functional electrical stimulation (FES). In this new device, a shape memory alloy (SMA) armature is used to perform the closing of the electrode. This technique makes the electrode installation around the nerve much easier, quicker, and safer. Both remarkable mechanical properties of SMA materials, namely, shape memory effect and superelasticity, can be used to obtain the desired mode of electrode closing. The fabrication procedure of the new electrode is described. It does not require any expensive or complex techniques. Bipolar and tripolar electrodes have been manufactured with an inner diameter of 1.6 mm and a cuff wall thickness of 0.8 mm. These electrodes are to be used for FES of the bladder in spinal cord injured patients. Acute studies in dogs are being carried out to validate the device and the implantation procedure. PMID- 10378927 TI - A custom designed chip to control an implantable stimulator and telemetry system for control of paralyzed muscles. AB - A custom designed chip has been developed for the control of paralyzed muscles. The system is capable of fulfilling the stimulus and telemetry needs of advanced functional neuromuscular stimulation (FNS) applications requiring multiple channels of stimulation and multiple channels for sensor or biopotential sensing. An inductive radiofrequency link provides power to the implant device as well as 2 way transcutaneous communication. An application specific integrated circuit (ASIC) decodes the commands and provides functional control within the implant, and modular circuitry provides specific implant functions. The ASIC chip provides up to 32 independent channels of stimulation with independent control of stimulus pulse duration, pulse amplitude, interphase delay, recharge phase duration, and pulse interval. It can also control up to 8 independent back telemetry analog channels with independent control of sampling rate and pulse powering parameters (amplitude and duration). The mixed analog digital chip has been fabricated in 1.2 microm n-well CMOS technology. PMID- 10378928 TI - Battery-powered implantable nerve stimulator for chronic activation of two skeletal muscles using multichannel techniques. AB - Chronic activation of skeletal muscle is used clinically in representative numbers for diaphragm pacing to restore breathing and for dynamic graciloplasty to achieve fecal continence. The 3 different stimulation techniques currently used for electrophrenic respiration (EPR) all apply high frequency powered implants. It was our goal to make these stimulation methods applicable for EPR by a battery-powered nerve stimulator that would maximize the patient's freedom of movement. Additionally, the system should allow the implementation of multichannel techniques and alternating stimulation of 2 skeletal muscles as a further improvement in graciloplasty. Generally, the developed implantable nerve stimulator can be used for simultaneous and alternating activation of 2 skeletal muscles. Stimulation of the motor nerve is achieved by either single channel or multichannel methods. Carousel stimulation and sequential stimulation can be used for graciloplasty as well as for EPR. For EPR we calculated an operating time of the implant battery of 4.1 years based on the clinically used stimulation parameters with carousel stimulation. The multichannel pulse generator is hermetically sealed in a titanium case sized 65 x 17 mm (diameter x height) and weighs 88 g. PMID- 10378929 TI - Use of functional electrical stimulation in the lower extremities of incomplete spinal cord injured patients. AB - After a program of therapeutic electrical stimulation, 3 groups of incomplete spinal cord injured (SCI) patients were identified, those in whom an improvement of both voluntary and stimulated muscle force was observed, those with an increase in stimulation response only, and patients in whom no effect of electrical stimulation training could be recorded. As it is difficult to predict the outcome of the electrical stimulation rehabilitation process, a diagnostic procedure was developed to predict soon after accidents which incomplete SCI patients are candidates for permanent use of a functional electrical stimulation (FES) orthotic aid. The candidates for chronic use of FES are patients with weak ankle dorsiflexors and sufficiently strong knee extensors. These patients are equipped with a single channel peroneal stimulator augmenting dorsiflexion and knee and hip flexion in a total lower limb flexion response. By applying FES to the ankle plantar flexors, the swing phase of walking can be significantly shortened and faster walking obtained. PMID- 10378930 TI - Demand for and use of functional electrical stimulation systems and conventional orthoses in the spinal lesioned community of the UK. AB - The use of and demand for functional electrical stimulation (FES) systems and conventional orthoses in the spinal cord lesioned population was assessed. The assessment was conducted by a postal survey of the members of the spinal injury associations in the U.K. Out of all the respondents, only 2% had used an FES system for walking. In comparison, 13% had used some kind of orthosis. Of the small numbers who had used an FES system for walking, more than half had no functional walking abilities. The majority of orthosis users had some independent walking ability. The demand for walking improvements was high among the respondents although this was not matched by the demand for improved orthotic solutions. In conclusion, it would appear that there is a need for simple FES systems offering walking improvement to the incomplete spinal cord lesioned (SCI) subject. PMID- 10378931 TI - Functional electrical stimulation and arm supported sit-to-stand transfer after paraplegia: a study of kinetic parameters. AB - The sit-to-stand transfer of paraplegic patients using functional electrical stimulation (FES) of the knee extensors and arm support was analyzed in the study. In a group of 8 completely paralyzed subjects who were trained FES users, kinematic and dynamic parameters were recorded during standing up trials. A contactless optical system was used to assess the human body motion. The forces acting on the human body were measured by multi-axis force transducers. On the basis of recursive Newton-Euler inverse dynamic analysis, the forces and torques acting on the body joints were calculated. The joint moments in the lower and upper extremities during the sit-to stand task are presented in this paper. The influences of the patient's strength, FES training duration, and rising strategy on the joint loading are discussed. PMID- 10378932 TI - Paraplegia: prolonged standing using closed-loop functional electrical stimulation and Andrews ankle-foot orthosis. AB - One T10 paraplegic male (CS) implanted in 1991 with a Nucleus FES-22 stimulator has been able to achieve closed-loop standing for 1 h. The knee angles are monitored by electrogoniometers, resulting in the quadriceps stimulation time being less than 10%. Stance stability is achieved by the Andrews anterior ankle foot orthosis (AFO). The use of accelerometers for trunk inclination and vertical acceleration during controlled stand-to-sit, diminishes slamming onto the seat. CS does one-handed tasks with objects of 2.2 kg. In another T10 paraplegic male (FR), surface stimulation was applied over 1.5 years to both femoral nerves at the groin for conditioning and prolonged standing. With quadricep conditioning, 55 Nm at 45 degrees of knee flexion is produced. With the AFO and knee monitoring, FR can stand uninterrupted for up to 70 min and perform one-handed tasks. In August 1998, he was implanted with the multifunctional Praxis FES 24-A stimulator for restoration of limb movements, bladder and bowel function, and pressure sore prevention. PMID- 10378933 TI - Stability and velocity in incomplete spinal cord injured subject gaits. AB - We have defined 2 indices describing gait kinematic and dynamic stability. We assessed their values in the gaits of 5 different paraparetic subjects. The indices are correlated to the gait velocity to prove the close relationship between overall gate velocity and stability. Based on stability analysis and certain kinematic parameters, some possible ways of increasing the average gait velocity are explained. PMID- 10378934 TI - Personal computer supported eight channel surface stimulator for paraplegic walking: first results. AB - Today functional electrical stimulation (FES) is used among other treatments to restore hand and arm function, to restore mobility of the lower extremities, for phrenic pacing, and in cardiomyoplasty. Common to all FES applications is that they require careful setup of stimulation parameters. To improve these tasks, personal computer (PC) based software for stimulation parameter evaluation and data acquisition was written. First, the described software was used to mobilize paraplegic patients in conjunction with an 12C bus controlled 8 channel surface stimulator. Electrodes were placed on each leg on the m. quadriceps and m. gluteus for hip and knee extension and the peroneal nerve to elicit flexion reflex. The fourth channel was used to correspond to subjects' individual needs. The stimulation patterns for standing up, walking, and sitting down easily could be set up and optimized by adjusting up to 128 stimulation parameters in a task specific way. PMID- 10378935 TI - MYOSTIM-FES to prevent muscle atrophy in microgravity and bed rest: preliminary report. AB - Long-term flights in microgravity cause atrophy and morphological changes of skeletal muscles. Training with mechanical devices is insufficient regarding the required time to exercise and space for devices. The objective of this project is to develop a passive training method based on functional electrostimulation (FES) to preserve muscle mass and fiber composition with minimal impairment to the cosmonaut. For a pilot experiment on the MIR space station, a suitable 8 channel FES device was developed. It consists of electrode trousers that carry surface electrodes and cables, 2 interconnected 4 channel stimulators, and a laptop personal computer (PC) for stimulator programming and processing compliance data. An automatic extensive training of 4 muscle groups of the lower extremities is performed for 6 h/day, with 1 s on and 2 s off tetanic contractions at 20-30% of maximum tetanic muscle force. The synchronous activation of antagonists of the thigh and lower leg prevents uncoordinated movements. PMID- 10378936 TI - Strength improvement of knee extensor muscles in patients with chronic heart failure by neuromuscular electrical stimulation. AB - Patients with severe chronic heart failure (CHF) suffer from marked weakness of skeletal muscles. Neuromuscular electrical stimulation (NMES) proved to be an alternative to active strength training. The objective of this study was to test the feasibility and effectiveness of NMES in patients with chronic heart failure. Seven patients (56.0 +/- 5.0 years, CHF for 20 +/- 4 months, left ventricular ejection fraction 20.1 +/- 10.0%) finished an 8 week course of NMES of the knee extensor muscles. The stimulator delivered biphasic, symmetric, constant voltage impulses of 0.7 ms pulse width with a frequency of 50 Hz, 2 s on and 6 s off. No adverse effects occurred. After the stimulation period, the isokinetic peak torque of the knee extensor muscles increased by 13% from 101.0 +/- 8.7 Nm to 113.5 +/- 7.2 Nm (p = 0.004). The maximal isometric strength increased by 20% from 294.3 +/- 19.6 N to 354.14 +/- 15.7 N (p = 0.04). This increased muscle strength could be maintained in a 20 min fatigue test indicating decreased muscle fatigue. These results demonstrate that NMES of skeletal muscles in patients with severe chronic heart failure is a promising method for strength training in this group of patients. PMID- 10378937 TI - Dynamic force responses in electrically stimulated triceps surae muscles: effects of fatigue and temperature. AB - To elicit dynamic force responses (unfused tetani) in isometric triceps surae muscles, low frequency electrical stimulation ranging from 12.5 to 30.0 Hz was applied. The fusing frequency (FF) and the relative dynamic force amplitude (DF) at the 20% and 40% maximum voluntary contraction (MVC) levels were calculated as parameters to determine effects of muscle fatigue (n = 6) and local muscle cooling. In the fatigued muscle (15 min plantar flexion at a 20% MVC level), the FF and DF increased when the fatigue was induced by voluntary contraction (FF increased from 19.6 to 22.5 Hz at 20% MVC) and also when induced by electrical stimulation (FF increased from 19.2 to 23.3 Hz). Cooling of the muscles showed an inverse effect on both parameters, indicating contractile slowing. The responsible physiological mechanisms as well as practical applications, using low frequency stimulation to monitor degenerative changes in muscles, are discussed. PMID- 10378938 TI - Clinical audit of 5 years provision of the Odstock dropped foot stimulator. AB - The Odstock dropped foot stimulator (ODFS) is a foot switch controlled single channel neuromuscular stimulator for correction of dropped foot. Following a randomized controlled trial, the ODFS was recommended for use in the United Kingdom's National Health Service and a clinical service established. The patient performance was assessed by measurement of walking speed over 10 m, physiological cost index (PCI), and by questionnaire. After 4.5 months stroke patients (n = 111) showed a mean increase in walking speed of 27% and reduction in PCI of 31% with stimulation and changes of 14% and 19%, respectively, unassisted. Multiple sclerosis patients (n = 21) gained similar orthotic benefit but no carry over. The principal reason cited for using the equipment was that it reduced the effort of walking. The principal reasons identified for discontinuing were an improvement in mobility, electrode positioning difficulties, and deteriorating mobility. A comprehensive clinical follow-up service is essential to achieve the maximum continuing benefit from FES based orthosis. PMID- 10378939 TI - Adaptive restriction rules provide functional and safe stimulation pattern for foot drop correction. AB - We report on our advances in sensory feedback data processing and control system design for functional electrical stimulation (FES) assisted correction of foot drop. We have applied 2 methods of signal purification on the bin integrated electroneurogram (i.e., optimized low pass filtering and wavelet denoising) before training adaptive logic networks (ALN). ALN generated stimulation control pulses, which correspond to the swing phase of the impaired leg when dorsal flexion of the foot is necessary to provide safe ground clearance. However, the obtained control signal contained sporadic stimulation spikes in the stance phase, which can collapse the subject, and infrequent broken stimulation pulses in the swing phase, which can result in unpredictable consequences. In this study, we have introduced adaptive restriction rules (ARR), which are initially used as previously reported and then dynamically adapted during the use of the system. Our results suggest that ARR provide a safer and more reliable stimulation pattern than fixed restriction rules. PMID- 10378940 TI - Standing up with denervated muscles in humans using functional electrical stimulation. AB - The use of electrical stimulation for denervated muscles is still considered to be a controversial issue by many rehabilitation facilities and medical professionals because prior clinical experience has shown that treating denervated muscle tissue using exponential current over a long time period constitutes an impossible task. Despite this fact, we managed to evoke tetanic contractions in denervated muscle using a long duration stimulation with anatomically shaped electrodes and sufficiently high amplitudes. The pulse amplitudes, which were being used for this purpose, exceeded by far the MED-GV and EC regulations (300 mJ/impulse). For this reason, an application has recently been submitted to have the EC regulations changed accordingly. It takes a tetanic contraction to achieve the desired muscle fiber tension, constituting a hypertrophic stimulus. It is also an appropriate means of exercise, which is capable of creating the metabolic and structural conditions needed (e.g, increased mitochondrial volume and capillary density) to obtain satisfactory muscle performance. With patients suffering from a complete spinal cord injury at level D12/L1, having motor and sensory loss in both lower extremities, we were able to train denervated muscle using long-duration stimulation, evoking single muscle contractions at first, soon followed by tetanic contractions against gravity. To increase the efficacy of this functional electrical stimulation (FES) strengthening program, we used ankle weights. With daily FES training over a period of 1-2 years, denervated muscle was exercised until it produced torques between 16 and 38 Nm in the m. quadriceps. With that muscle force, it is possible to stand up from a sitting position in parallel bars. Our results show that denervated muscle in humans is indeed trainable and can perform functional activities with FES. Furthermore, this method of stimulation can assist in decubitus prevention and significantly improve the mobility of paraplegics. PMID- 10378941 TI - Computer simulation of field distribution and excitation of denervated muscle fibers caused by surface electrodes. AB - In the course of this study, 2 submodels have been developed and combined, the 2 D finite element modeling of the electrical potential distribution in the human thigh and a Hodgkin and Huxley (HH) type model to calculate fiber excitation and action potential propagation. To determine the excitation of the target muscle fiber with the help of the activating function, the fiber's orientation within the muscle has to be known. The electric field along the fiber has to be calculated as a function of the applied electric current and the potential at the electrodes, respectively. The excitement of the muscle fibers varies across a wide range depending on the active and passive membrane parameters and the intracellular and extracellular mediums. Persisting denervation leads to a decay of muscle cells, and a partial substitution by fibroblasts occurs. The electrical activation of these tissues is more difficult, and biphasic stimulation pulses up to 200 ms in duration and 60-100 V in amplitude are needed to cause a contraction of the denervated muscle. An example shows the field distribution and the simulated activity in one representative muscle fiber of a well trained m. rectus femoris. PMID- 10378942 TI - Long pulse biphasic electrical stimulation of denervated muscle. AB - In recent years a number of studies have employed long pulse biphasic stimulation as a treatment for denervated muscle to improve tissue quality and in some cases to improve contractile capability sufficient to restore function. However, in the U.K., this treatment is yet to be widely adopted clinically. A 5 subject, case based pilot study of long pulse biphasic direct stimulation of peripheral limb denervated muscle is being conducted and its effect on the tissue evaluated by measurement of muscle bulk, limb blood flow, and skin temperature. In cases of partial denervation. trapezoidal shaped pulses are used to minimize sensory and motor nerve fiber recruitment. PMID- 10378943 TI - Experimental wound healing with electrical stimulation. AB - The effect of alternating current (AC) and direct current (DC) stimulation was studied on experimental pressure ulcer healing in a new monoplegic pig model. The study was conducted in 30 healthy young Hanford minipigs. The rate of wound healing, histology, vascularization, collagen formation, microbiology, perfusion, and the mechanical strength of the healed wounds were studied. Normal pigskin was compared to denervated control and denervated AC and DC stimulated healed skin. Hind limb denervation was by right unilateral extradural rhizotomies from the L2 to S1 nerve roots. Reproducible uniformly controlled Stage III or higher tissue ulcers were created. When compared to the control wounds, both the AC and DC stimulated wounds showed reduced healing time and increased perfusion in the early phases of healing. DC stimulation reduced the wound area more rapidly than AC, but AC stimulation reduced the wound volume more rapidly than DC. The electrical stimulation did not reduce the strength of the healing wounds below those of the nonstimulated controls. The applied current appears to orient new collagen formation even in the absence of neural influences. PMID- 10378944 TI - Three-dimensional changes in the upper airway during neuromuscular stimulation of laryngeal muscles. AB - During swallowing, airway protection depends upon adequate glottal closure and laryngeal elevation to prevent the entry of substances into the airway. Three dimensional changes in the upper airway during laryngeal muscle stimulation in a canine model were quantified in animals implanted with Peterson type stimulating electrodes in the inferior and superior portions of the thyroarytenoid muscle, together with a reference electrode. Computer tomography scanning was performed on an IMATRON scanner with a 3 mm slice thickness advanced at overlapping 1 mm increments. Stimulation of the thyroarytenoid muscle produced adductions of the vocal fold towards the midline and changes in the supraglottic region as well as the glottis; the glottic wall was compressed medially above and below the glottis. These results suggest that chronic neuromuscular stimulation can effect glottic protection by reducing the glottal opening and may be beneficial for patients with central control disorders affecting airway protection during swallowing. PMID- 10378945 TI - Electromyogram-controlled functional electrical stimulation for treatment of the paralyzed upper extremity. AB - Spinal cord lesions at level C5 to C6 lead to loss of hand functions and lesions at C4 to additional deficits of arm functionality. The presented dual channel surface stimulator with dual channel electromyogram (EMG) measurement was developed to investigate control strategies for an EMG-controlled implantable stimulation system and serves in addition as a therapy device for patients with partial innervation but weak muscle force. Four different control strategies for stimulation amplitude are available. The amplitude can be preset manually or can follow the preprocessed EMG signals proportionally. The shoulder control program allows proportional control of both stimulation channels with one EMG channel while the second EMG channel serves as the channel selector. Finally, a special feedback training program triggers a stimulation burst when EMG activity is detected. During a 2 year patient study, 18 patients from 2 hospitals and 1 rehabilitation center performed the feedback training. Almost all patients obtained an improvement of functionality. Apart from muscle strengthening, the feedback effect led to an improvement of proprioception and supported relearning of motions. For the documentation of the training status, functional muscle test (British Medical Research Council) and measurements of power, angle, torque, muscle fatigue, and EMG were performed. Obviously, EMG triggered stimulation provides several advantages compared to conventional passive electrical stimulation. PMID- 10378946 TI - Functional control of the hand in tetraplegics based on residual synergistic EMG activity. AB - A microprocessor controlled device (MeCFES) was used for the investigation of the possibility of restoring hand function in C5 tetraplegics with paralysis of the hand. To date, 3 tetraplegics have been testing the system. The myoelectric signals from wrist extension were recorded and used as control signals for functional electrical stimulation (FES) of thumb adduction/flexion. The results have shown that the device can improve the hand function of tetraplegics. In this part of the work, a hand function test was designed and used to assess the results. PMID- 10378948 TI - Introduction PMID- 10378949 TI - Anxiety in the medically ill: An overview. PMID- 10378947 TI - New peptides prevent brain damage. PMID- 10378950 TI - Anxiety in the medically ill: nosology and principles of differential diagnosis. AB - Anxiety and anxiety disorder have been estimated to occur in 5% to 20% of medical inpatients and 4% to 14% of medical outpatients. These estimates do not distinguish between anxiety symptoms and anxiety disorders, nor between the various causes of anxiety in the medically ill. This article reviews the epidemiology, nosology, disorders in the differential diagnosis, and principles of differential diagnosis of anxiety in the medically ill. A widely endorsed nosology for anxiety disorders caused by medical illness and its treatments is a relatively recent development. General guidelines for differentiating between specific diagnoses in the differential diagnosis of anxiety in the medically ill are now provided in the Diagnostic and Statistical Manual of Mental Disorders, 4th Edition, but, as is often the case, it is easier to describe the distinctions in theory than it is to make them in actual practice. PMID- 10378951 TI - Anxiety and endocrine disease. AB - It is our goal in this article to review the literature and define those endocrinological diseases that often include anxiety states as part of their initial presentation or as a characteristic symptom seen during their course. Understanding the mechanism by which anxiety develops as a routine part of these neuroendocrinological disorders may help us understand the organic basis of anxiety disorders. Research using new neurochemical, neuroanatomical, and brain imaging techniques may further define the structural and physiological underpinnings of the anxiety disorders. PMID- 10378952 TI - Anxiety in patients with pulmonary disease: comorbidity and treatment. AB - Anxiety is a common and sometimes disabling symptom among patients with respiratory disease. Anxiety disorders appear to be the most prevalent psychiatric disorders in clinical samples of patients with pulmonary disease. Recognition that the differential diagnosis of dyspnea and anxiety includes both pulmonary and psychiatric conditions can be crucial to appropriate medical management and minimizing iatrogenic harm. This article reviews the epidemiology, comorbidity, diagnosis, and treatment of anxiety syndromes in patients with pulmonary disease. Successful treatment of anxiety disorders can substantially improve quality of life and a variety of treatment options are available. Safe and effective pharmacotherapy requires attention to potential adverse drug effects on pulmonary function and drug-to-drug interactions. Nonpharmacological treatments such as cognitive/behavioral therapies offer effective treatment without the risk of medication side effects. PMID- 10378953 TI - Anxiety and neurological disorders. AB - Anxiety disorders occur quite frequently in patients who have neurologic disorders. In fact, several studies show that anxiety disorders occur more frequently than mood disorders in patients with neurologic conditions. Despite high prevalence rates, investigators have neglected the study of anxiety disorders in these patients. Two major reasons for the neglect are the diagnostic hierarchy and exclusion criterion found in Diagnostic and Statistical Manual (DSM)-III-R and DSM-IV. This article discusses problems in the diagnosis of anxiety disorders in patients with neurologic conditions and presents a brief literature review of the relationship between anxiety and neurologic disorders. In addition, the authors describe the differential diagnosis of anxiety in neurologic conditions and treatment options. PMID- 10378954 TI - Anxiety in patients with multiple sclerosis. AB - Anxiety disorders are quite common, and frequently overlooked, in patients with Multiple Sclerosis (MS). This is often due to the difficulty differentiating anxiety from personality correlates or reactive tendencies in patients with neurologic disease. This chapter offers the consulting psychiatrist guidelines for providing psychological support to patients with MS at various stages of their disease. DSM-IV-based differential diagnosis, psychotherapeutic techniques, behavioral interventions, and pharmacological support (including the newer alternative therapies) are reviewed. The physical, functional, and symbolic losses caused by this chronic and progressive disease are considered in the broader context of individual patients' lives. Particular attention has been given to specific pharmacological treatment of steroid-induced anxiety. This is essential knowledge for the consulting psychiatrist. The overlap between depressive symptoms, manic symptoms and cognitive changes in MS patients is reviewed with special emphasis on the structural correlates. Current neuro imaging techniques, including emerging technologies such as gadolinium enhancement, single photon emission computed tomography and brain electrical mapping (BEAM), now provide a far more accurate view of brain damage in MS. This permits diagnosis of the disease much earlier, and is also beginning to show correlation between neuropsychiatric clinical findings, and the nature and location of demyelinating plaques in the brains of MS patients. This chapter seeks to clearly define the associations between anxiety disorders and cerebral involvement in MS patients, suggesting that common neurological and biochemical mechanisms are more extensive than generally suspected. It is hoped that this information will aid clinicians in more accurately diagnosing and effectively treating anxiety in MS patients. PMID- 10378955 TI - Anxiety in patients with cancer and human immunodeficiency virus. AB - Physicians who treat patients with cancer or human immunodeficiency virus (HIV) infections frequently encounter the psychological and physical manifestations of anxiety in these populations. This chapter provides a review of the prevalence of anxiety disorders in cancer and HIV patients. The problems in assessment and diagnosis of anxiety in these patients are discussed, and a literature review of the types of anxiety disorders commonly identified in the context of cancer and HIV/AIDS is presented. Finally, the treatment of anxiety in cancer and HIV patients utilizing both pharmacologic and nonpharmacologic modalities is reviewed. PMID- 10378956 TI - Pharmacological treatment of anxiety in the medically ill patient. AB - Treatment guidelines which are available are intended for treatment of medically healthy patients. There is little information in the literature regarding the treatment of patients who are medically ill. Anxiety and anxiety disorders are frequently encountered in patients with serious medical disorders. These medically ill patients may have substantial changes in pharmacokinetic factors as a result of their current illness or treatment. Also, treatment considerations related to their accompanying medical treatment(s) are sometimes required. These various factors may result in a different initial choice of agents, alterations in the dosage of a given agent, or even preclude the use of some standard agents. In this chapter, we first present a review of the current psychopharmacologic treatment of anxiety disorders. In the second section, we review the potential pharmacokinetic consequences of serious hepatic renal, pulmonary and cardiac disease relevant to the pharmacological treatment of anxiety. PMID- 10378957 TI - Cognitive/behavioral therapy of anxiety in the medically ill: cardiac settings. AB - Anxiety appears to be a strong risk factor for ischemic heart disease (IHD) and specifically fatal IHD. However, no randomly assigned, controlled, clinical trial targeting anxiety yet exists demonstrating an impact on objective cardiac outcomes. Situational anxiety is frequent in cardiac populations and can diminish quality-of-life by increasing symptoms/disability and result in unnecessary medical system utilization. "Noncardiac" chest pain is common both in patients with objective coronary disease and in patients whose cardiac workup is negative. Both presentations of chest pain respond to cognitive/behavioral therapy, and imipramine has been found to be effective for chest pain unaccompanied by coronary disease. Because anxiety-like symptoms overlap with symptoms of IHD (eg, chest pain, dyspnea, dizziness, palpitations) and can be caused by organic factors, the diagnosis and treatment of anxiety in these populations require special considerations. PMID- 10378958 TI - Adjuvant therapy of breast cancer. Introduction. PMID- 10378959 TI - Radiation therapy as an adjunct to primary surgery in early stage breast cancer: biological and clinical rationale. AB - In spite of the widespread use of adjuvant endocrine and cytotoxic chemotherapy, a substantial proportion of patients with early-stage breast cancer eventually develop a distant disease recurrence. Local control also remains a clinically significant problem in subsets of patients. Whether improved local control through the use of postoperative radiation therapy would prevent distant dissemination has been much debated for several decades. Studies on the long-term outcome of systemically untreated breast cancer patients indicate that breast cancer in many patients is a local disease that can be cured by surgery or radiotherapy. Randomized trials of breast screening have also shown that a delay in effective local treatment is associated with an increased incidence of distant dissemination and death from to the disease. Data from individual randomized trials and overviews of postoperative radiation therapy have indicated that radiation therapy as an adjunct to primary surgery is associated with a decrease in distant dissemination and breast cancer death. This benefit may be translated into a substantial overall survival benefit, provided that the treatment technique avoids long-term cardiac side effects. In many of the older radiation therapy trials, such effects appear to have balanced the benefit in terms of a reduced incidence of distant disease among the patients allocated to radiotherapy. PMID- 10378960 TI - Locoregional failure rates in patients with involved axillary nodes after mastectomy and systemic therapy. AB - Published series vary substantially in describing the incidence of locoregional failure after mastectomy among patients with involved axillary lymph nodes who receive systemic therapy. There are few data on such risks with regards to particular patient subsets (such as those defined by combinations of tumor size and nodal status). This article reviews the available data on these subjects as well as problems in their interpretation and clinical use. PMID- 10378961 TI - Postmastectomy radiation in patients with one to three positive axillary nodes receiving adjuvant chemotherapy: An unresolved issue. AB - The rationale for postmastectomy radiation is based on the prevention of locoregional recurrence in the chest wall, regional lymphatics, or both. The randomized trials of postmastectomy radiation in patients with one to three positive nodes receiving adjuvant chemotherapy have shown a proportional reduction in locoregional recurrence rates of two thirds. The absolute benefit, however, varies with the magnitude of the risk in patients who do not receive radiation. The survival benefit from radiation is best explained by the prevention of an isolated locoregional recurrence, which could serve as a source of fatal distant metastases and parallels the difference in the total incidence of distant metastases. The current dilemma is to identify patients with one to three positive nodes who have had an adequate axillary dissection and remain at substantial risk for a locoregional recurrence despite adjuvant chemotherapy. The routine use of postmastectomy radiation in all axillary node-positive patients requires further evaluation. PMID- 10378962 TI - Role of postmastectomy radiotherapy: A medical oncology perspective. AB - Current practice in the management of patients who undergo a mastectomy does not usually include radiotherapy. Data from both meta-analyses and two randomized studies challenge this approach. Part of the skepticism about postmastectomy radiotherapy is what biological rationale would justify this intervention. Hellman has proposed the spectrum hypothesis, which explains the potential indication for additional local therapy for such patients. Another concern is the risk for increased toxicity, especially cardiotoxicity in patients who receive anthracycline-based adjuvant regimens. The addition of new agents (eg, Herceptin) also requires careful monitoring with regard to toxicity. Furthermore, the timing of radiation with chemotherapy is problematic. Overall, there is now evidence to support a role for postmastectomy radiotherapy, but further studies are needed on how best to incorporate this modality in multimodality treatment of early-stage breast cancer. PMID- 10378963 TI - Postmastectomy irradiation: rationale for treatment field selection. AB - The goal of postmastectomy irradiation is to eliminate residual viable tumor in tissue remaining after standard mastectomy. Because this subclinical disease is, by definition, not detectable by current technology, the choice of patients and treatment volumes for postmastectomy irradiation must be inferred from a variety of data sources. The absolute risk of locoregional recurrence is related to the stage of disease, the extent of lymphatic involvement, and other treatment received. Patterns of failure analyses consistently identify the chest wall as the most important target for treatment with radiation therapy in high-risk patients. When patients with multiple locoregional sites of recurrence are included, the chest wall may be involved in as many as 60% to 80% of patients. The second most common place for locoregional failure is the undissected lymphatics of the paraclavicular region. The cumulative probability of failure in this region ranges from 10% to 35% of the patients treated for locoregional recurrence. Microscopic tumor metastases in the internal mammary chain are theorized to represent a potential source for distant metastases. Each of the prospective trials of postmastectomy irradiation that have shown survival benefit included the internal mammary chain within their target volume. Nonetheless, local failure in the internal mammary nodes is an uncommon finding. Similarly, after a level I and II axillary dissection, axillary failure is a minor component of local recurrence risk, and it is probable that only a subset of patients may benefit from axillary irradiation. PMID- 10378964 TI - Adjuvant irradiation after mastectomy in women with one to three positive axillary nodes: then no; now yes. AB - The decision whether to offer women with one to three positive nodes postmastectomy radiation after adjuvant chemotherapy has been the subject of intense discussion since the publication of two major randomized prospective trials. Although radiotherapy after mastectomy was an established treatment for women with four or more positive axillary nodes, before these studies, existing data did not justify its use in patients with less extensive nodal involvement. Now with results from these studies showing improved survival after radiotherapy in all node-positive premenopausal and perimenopausal women, with perhaps its greatest benefit in women with one to three positive nodes, practice patterns are again shifting toward strong consideration of treatment in women with less tumor involvement. The arguments supporting this new treatment philosophy are presented. PMID- 10378965 TI - Postmastectomy radiotherapy: toxicities and techniques to reduce them. AB - The role of locoregional radiation therapy after mastectomy is controversial. It reduces the risk of tumor relapse, improves breast cancer-specific survival and possibly overall survival, but has potential morbidity. This article reviews the technical aspects of postmastectomy radiation therapy and its associations with treatment-related morbidity. We consider common problems that arise in the technical setup of radiation fields. Adverse effects of postmastectomy radiation therapy may be reduced or prevented by careful radiation treatment planning. PMID- 10378966 TI - Postmastectomy radiation therapy: A surgical perspective. AB - Controversy exists about whether postmastectomy radiotherapy prolongs survival or is merely of benefit to maintain local control. Surgical series of patients treated by radical or modified radical mastectomy alone show locoregional failure rates ranging from 4% to 26%. The likelihood of involvement of supraclavicular and internal mammary nodal metastases increases as the extent of axillary nodal involvement increases. Adjuvant systemic therapy appears to have limited impact on the incidence of locoregional failure. Untreated nodal disease is a potential source of tumor dissemination and provides a biological rationale for a survival benefit for irradiation. PMID- 10378967 TI - Postmastectomy radiotherapy: randomized trials. AB - Postmastectomy radiotherapy decreases threefold the risk of locoregional recurrences according to the results of many randomized trials and overviews. This risk is mainly related to the number of involved axillary nodes (ie, about 25%, 35%, and 55% at 10 years when 1 to 3, 4 to 9, and 10 or more nodes are involved). In contrast, at 10 years, fewer than 15% of patients with negative axillary nodes relapse locally. The effect of postmastectomy radiotherapy on distant metastases and overall survival is a controversial issue. On the one hand, results are compatible with the existence of a mechanism of secondary dissemination generated from locoregional tumor nests. The beneficial effect of radiotherapy may be observed in the absence or presence of adjuvant systemic treatment. On the other hand, a deleterious late toxic, mainly cardiac, effect of radiation has also been shown. This point emphasizes the importance of radiation technique and quality to obtain a positive balance in terms of overall survival. PMID- 10378968 TI - Is the use of radiation therapy after mastectomy cost-effective? AB - With the publication of two randomized trials showing an improvement in overall survival after the use of postmastectomy radiation therapy, interest in the use of radiation therapy in this setting has been rekindled. These results are in contrast to those reported in the most recent meta-analysis of the Early Breast Cancer Trialists' Collaborative Group, in which a statistically significant survival benefit was not detected. Although evidence of a survival benefit was sufficient in the past for an intervention to gain acceptance, payers are increasingly interested in knowing whether its use is also cost-effective. This article briefly reviews the methods used in performing cost-effectiveness analyses, summarizes the results of one published and a second preliminary cost effectiveness analysis of postmastectomy radiation therapy, and highlights several areas for future research. PMID- 10378969 TI - Overview of randomized trials in high risk breast cancer patients treated with adjuvant systemic therapy with or without postmastectomy irradiation. AB - Postmastectomy irradiation has been analyzed in several randomized trials, usually without the presence of systemic therapy. Overview analysis of these studies has shown that, although postmastectomy irradiation generally reduces the locoregional recurrence rate by a factor of 3, this is not transferred into a general long-term survival benefit. Recently, a clinically relevant survival gain from adjuvant radiotherapy has been shown in high-risk premenopausal patients also treated with adjuvant chemotherapy. There are now 11 published trials, including more than 6000 patients, in which the effect of postmastectomy radiotherapy has been evaluated in the presence of adjuvant systemic therapy. This article reviews the available data from these trials, as well as problems in their design, sample size, inclusion criteria, quality, and extent of treatments, and length of follow-up time. PMID- 10378970 TI - Postmastectomy radiotherapy: future directions. AB - With careful interpretation of existing studies of postmastectomy radiotherapy, much has been learned about the ability of radiotherapy to significantly reduce local failure and potentially impact on survival. With this knowledge, however, has come additional questions about the mechanisms by which radiotherapy could affect systemic control and the extent of that benefit. Therefore, these questions need to be investigated in well-designed, randomized trials that incorporate aggressive surgical techniques and contemporary chemotherapy regimens into the clinical plan. A trial that is currently in progress should give additional insight into whether regional irradiation in the modern era, which incorporates the internal mammary nodes in the radiotherapy field, impacts systemic control. An upcoming trial will investigate whether women at moderate risk for locoregional failure will benefit from comprehensive radiotherapy after aggressive surgery and chemotherapy. And, although no national studies are currently planned to test the optimal sequencing of radiotherapy and chemotherapy, consideration should be given to studying this issue in large, randomized trials. PMID- 10378971 TI - Special populations in occupational health. AB - This introductory chapter lays the groundwork for the in-depth examination of special populations that follows. Drs. Frumkin and Pransky discuss what makes a population special, health disparities over the generations, and the evolving recognition of special populations in occupational health. PMID- 10378972 TI - The economic and social context of special populations. AB - Changes in both technology and international trade are altering the world economy and hence are affecting the demand and supply of labor and the nature of work and working conditions. New materials, faster and more powerful computers, electronic and mobile communications, alternative energy systems, miniaturization, robotics, and biotechnology pose new opportunities, problems, and challenges. The tremendous expansion in information-based technologies in both manufacturing and services has resulted in impressive increases in productivity and demand for new skills, but they also have brought about the displacement and de-skilling of some labor by capital, the lowering of wages, and the increase of contingent, part time, and temporary work. Special populations, in particular, may be differentially impacted. PMID- 10378973 TI - Minority workers and communities. AB - Environmental and occupational hazards do not affect all communities equally. Members of ethnic and racial minorities, whether as working people or as community residents, sustain disproportionate risks from chemical, physical, and biological hazards. This paper reviews the nature of these disproportionate risks, focusing primarily on the workplace, but considering general environmental exposures as well. It discusses three principal mechanisms of increased risk: excessive hazardous exposures in both the workplace and the general environment, increased susceptibility, and inferior healthcare. It presents evidence that, as the result of these factors, members of minority groups display elevated rates of work-related illness, injury, fatality, and disability. Finally, it offers recommendations with regard to research, primary prevention, minority recruitment into the occupational health professions, and treatment and compensation for injured and ill minority workers. PMID- 10378974 TI - Young workers. AB - Until recently, today's occupational safety and health experts have paid little attention to safety and health concerns of working youth. Yet with millions of children and adolescents employed each year, young workers are indeed a special population at risk deserving special attention. Occupational safety and health professionals have critical knowledge and skills to contribute to researching special issues for young workers and promoting safe and healthful work for youth. Unique opportunities for intervention hold the potential for new and rewarding partnerships with, for example, pediatricians and adolescent health specialists, child labor regulators, child injury prevention professionals, maternal and child health professionals, educators, and community leaders. Lessons learned in targeting young workers can have important implications for reaching other special populations that have not been well addressed through conventional approaches to occupational safety and health. PMID- 10378975 TI - Older workers. AB - As the population ages, there is increasing attention to the occupational health of older workers and the relationship between work and aging. There are both positive and negative factors that characterize differences between older and younger workers. Some of these are well documented, but many are based on stereotypes about competence, knowledge, and work capacity. Workers meet the demands of work through the use of a combination of resources, including physical, mental, and social capacities as well as motivation and experience. While older workers do have decreased physical capacities and somewhat slower mental processing, motivation and expertise can provide important balance. Factors that make advancing age into a handicap are mostly related to working conditions that impose constraints that outstrip actual human capabilities and work organization that denies employees growth in their jobs. PMID- 10378976 TI - Women workers: the social construction of a special population. AB - This paper presents data on the employment characteristics of women workers in the United States, together with a discussion of the biases that exist in current employment recording systems. These biases lead to an undercounting of women workers and an underestimation of risks related to both domestic and paid employment. The paper delves into the inappropriateness of considering women workers as a "special" category of workers. Also covered are the occupational health and safety hazards that women face on the job, with associated morbidity and mortality, and the relationships between women's work and women's health. This analysis presents ideas about research and policy needs in the area of women's occupational health. PMID- 10378977 TI - Workers with disabilities. AB - Individuals with disabilities constitute a sizable portion of the workforce and represent the majority of working-age persons who are unable to work. Historically, barriers to employment have included attitudinal discrimination by employers, lack of workplace accommodations, and inadequate job training. The disability rights movement has achieved considerable success in promoting legislation to remove these barriers and uphold equal employment. Research suggests that many employers actively attempt to incorporate persons with disabilities into the workforce and gain substantial economic benefit from their participation, without incurring burdensome expenses. Occupational health providers are asked by employers and others to provide input on feasibility and safety, a difficult task given the lack of scientific study on the occupational abilities and risks associated with specific disabilities. Providers have an important role in promoting the equal employment of disabled persons, by providing objective opinions on their ability and risks on the job and suggesting workplace accommodations and treatments that enhance the ability to work. PMID- 10378978 TI - Genetically and medically susceptible workers. AB - The likelihood of an individual becoming ill from a hazardous material or condition is strongly influenced by both their genetic makeup and their underlying state of health. Although the past decade has seen great advances in understanding human variation in health and genetic polymorphisms and in the diagnosis and treatment of disease, much less progress has been made in effectively using this information to protect worker health. Scientific evidence for increased susceptibility often is weak and rarely satisfies legal thresholds for sufficient risk to warrant exclusion from a particular job. When public safety is a major concern, many legally mandated exclusions are not well justified. Medical opinions about fitness to work should be based upon a systematic and credible analysis of the condition, its relationship to ability and risk for a particular job, and knowledge of possible accommodations. Conclusions should reflect the limitations of scientific knowledge and guidance from antidiscrimination legislation. PMID- 10378979 TI - The occupational health status of hired farm workers. AB - The U.S. hired farm work force presently is two-thirds foreign-born: mostly young Mexican men with low educational attainment who neither read nor write English. Sixty percent earn so little that they and their families live in poverty. Four of ten migrate to find work, 33% are not authorized to work in the U.S., and 25% work for a labor market intermediary, usually a labor contractor. Few hired farm workers have health insurance of any kind and, despite low incomes, relatively few seek or receive government benefits. Government regulation of the workplace exempts agricultural employers from numerous provisions that apply to other industries; for example, agriculture is exempt from portions of the Fair Labor Standards Act, allowing children as young as age 12 to work in the fields, and employers with 10 or fewer employees are exempt from OSHA regulation. Only 12 states require farm employers to carry workers' compensation insurance. While hired farm workers face significant safety and health risks, there are major gaps in existing research covering this occupational group. An ad hoc task force convened by NIOSH developed a prioritized agenda for occupational safety research in this population: musculoskeletal disorders, pesticide-related conditions, traumatic injuries, respiratory conditions, dermatitis, infectious diseases, cancer, eye conditions, and mental health. PMID- 10378980 TI - Across the water and down the ladder: occupational health in the global economy. AB - As the world economy becomes more integrated, and as industrial production expands in poor nations, workers in these nations face a range of occupational health and safety hazards. This article discusses the political economy of occupational health in developing nations by reference to multinational companies, free trade zones, free trade agreements, and the export of hazards. It reviews the special circumstances of occupational safety and health in developing nations and presents data on morbidity and mortality related to workplace exposures in these nations. Finally, it discusses approaches to improving workplace safety in developing nations, including policy initiatives, both mandated and voluntary, and public health initiatives, including training, technical assistance, collaborative research, and advocacy. PMID- 10378981 TI - Legal solutions for the problems of special populations at risk. AB - Some populations of workers face competitive disadvantages in the labor market and increased risks at work. This paper discusses three areas of law that are relevant to the occupational health concerns of these subgroups of workers. First, laws against discrimination in employment on the basis of race, sex, age, and disability are described. Second, laws that provide universal protection for all workers, including regulatory laws (e.g., OSHA) and social insurance programs (e.g., workers' compensation) are evaluated in terms of the extent to which they provide effective protection for vulnerable populations. Third, the nature of legal protections for people at specific increased risk for disease is assessed. Appendices provide specific summaries of relevant laws. PMID- 10378982 TI - Organizing marginalized workers. AB - Figures from the U.S. Department of Labor show that low-wage or marginalized workers are more likely to be injured on the job and suffer more work-related medical conditions than better-paid workers. Despite an increasingly hostile organizing climate, market globalization, and corporate downsizing, significant progress has been made in organizing marginalized workers. A multifaceted, comprehensive organizing strategy, incorporating union-building strategies that include (but are not limited to) safety and health, must be used by unions to successfully organize marginalized workers and obtain the first contract. PMID- 10378984 TI - EDITORIAL. PMID- 10378985 TI - Leukotrienes, leukotriene receptor antagonists and leukotriene synthesis inhibitors in asthma: an update. Part I: synthesis, receptors and role of leukotrienes in asthma. AB - Asthma is a chronic inflammatory disease associated with airflow obstruction. Airflow obstruction results from contraction of airway smooth muscle, mucosal oedema, increased secretion of mucus and infiltration of the airway wall by inflammatory cells, particularly eosinophils. Leukotrienes are thought to contribute to the pathophysiology of asthma. Leukotrienes are synthesised from arachidonic acid by a specific synthesis pathway whose key enzyme is 5 lipoxygenase. Cysteinyl leukotrienes (leukotrienes C4, D4 and E4) have been shown to mimic all the pathologic changes that are characteristic of asthma, whereas leukotriene B4 does not appear to exert biological properties relevant to asthma. Cysteinyl leukotrienes bind to two receptor subtypes: CysLT1 and CysLT2. Most of the biological properties of cysteinyl leukotrienes relevant to asthma are mediated through CysLT1 receptor stimulation. PMID- 10378986 TI - Leukotrienes, leukotriene receptor antagonists and leukotriene synthesis inhibitors in asthma: an update. Part II: clinical studies with leukotriene receptor antagonists and leukotriene synthesis inhibitors in asthma. AB - The demonstration that leukotrienes, mainly cysteinyl leukotrienes, have biological properties relevant to the pathogenesis of asthma has stimulated the development of many therapeutic compounds to block these deleterious effects. Two main classes of leukotriene modulators have been developed: CysLT1 receptor antagonists and leukotriene synthesis inhibitors. This article reviews the pharmacodynamics, the effects on baseline airway function, the protective effects in airway challenges as well as the results in chronic asthma of the different leukotriene modulators. In addition, the complementary anti-inflammatory effect of leukotriene modulators to that of corticosteroids and H1-histamine receptor antagonists is reviewed. Finally, a concise overview of the clinical responsiveness to this new class of drug, the safety and the drug interactions as well as the place in the strategies of treatment for asthmatic patients of the leukotriene modulators is also provided. PMID- 10378987 TI - The evaluation of the antianaphylactic effect of Oryza sativa L. subsp. hsien Ting in rats. AB - We studied the effect of the methanol extract of Oryza sativa L. subsp. hsien Ting (OSHT) on anaphylaxis. OSHT (0.001-1.0 mg g-1body weight (BW)) dose dependently inhibited systemic anaphylaxis induced by compound 48/80 in rats. When OSHT was pretreated at concentrations ranging from 0.001 to 1.0 mg g-1BW, the serum histamine levels were reduced in a dose-dependent manner. OSHT (0. 001 1.0 mg g-1BW) also inhibited local anaphylaxis activated by anti-dinitrophenyl (DNP) IgE. Moreover, OSHT dose-dependently inhibited the histamine release from rat peritoneal mast cells (RPMC) activated by compound 48/80 or anti-DNP IgE. The level of cAMP in RPMC, when OSHT was added, significantly increased approx. 20 fold compared with that of basal cells. These results indicate that OSHT possesses strong antianaphylactic activity by inhibition of histamine release from mast cells in vivo and in vitro. PMID- 10378988 TI - Effect of verapamil on responses to endothelin-1 in aortic rings from streptozotocin-induced diabetic rats. AB - In this study, we investigated the constrictor responsiveness to endothelin-1 (ET 1, 10-30 n m) of aortic rings (under 1 g resting tension in Krebs-Bicarbonate solution) from 8-weeks streptozotocin (STZ, 65 mg kg-1, i.p)-induced diabetic rats and vehicle-treated control rats. The maximum ET-1-induced contraction of the aorta in diabetic rats was increased by 150%, but the EC50 of ET-1 remained unchanged. Although in both groups, verapamil reduced the constrictor responses to ET-1 (diabetic group P<0.001, control group P<0.05), there were not any significant differences between PD2 values. These results suggest that verapamil inhibits ET-1-induced Ca2+ entry through the L-type channel and this effect did not change in diabetes mellitus. PMID- 10378989 TI - Spontaneous reporting of adverse drug reactions in elderly patients in Sicily (Italy). AB - The aim of this study was to investigate the spontaneous reports of suspected adverse drug reactions, observed in elderly patients (over 65 years of age) in Sicily (Italy) during the period from 1 January 1995 to 31 December 1997. Of 1307 reports, the geriatric ADRs were 284 (21.7%); 92 (32.4%) of these were serious. There was a correlation between the reporting rates of ADRs and the increase of age. Similar trends are seen in the number of serious ADR reports. Old-older patients result most frequently affected by serious ADRs. The antimicrobial and musculo-skeletal drugs were responsible for 48. 3% of the whole suspected geriatric ADRs. The commonest ADRs reported for the elderly, affected skin and the gastrointestinal system. There was probably a correlation between multi-drug consumption, comorbidity and seriousness of ADRs. A higher percentage of serious ADRs originated from university hospitals (57. 1%). PMID- 10378990 TI - Prescribing habits of general practitioners in choosing an empirical antibiotic regimen for lower respiratory tract infections in adults in Sicily. AB - The survey was carried out, between September 1995 and May 1996, in order to describe the prescriptive behaviour among Sicilian general practitioners (GPs) in choosing an empirical antibiotic regimen for LRTIs in adult patients and begin an educational process which involves the same GPs in decisions regarding their prescriptions and in performing local guidelines. Each practitioner filled out a questionnaire for each therapeutic intervention which ended with an antibiotic prescription. The questionnaire also enquired into the patient's characteristics, diseases to be treated and drug prescription. Doctors were asked to give an opinion about the severity assessment of the infectious disease before choosing the antibiotic treatment, in order to evaluate the prescriptive behaviour of physicians related to the patient's symptoms. Of all Sicilian GPs approached, 76 physicians from 25 Sicilian towns, with a patient population of 96,630, agreed to participate. The GPs used 49 different molecules and six different associations of two antibiotics. The most frequently used antibacterial agents were cephalosporins (55.0%). Penicillins (11.7%), fluoroquinolones (11. 4%), macrolides (10.1%) and combinations of penicillins with beta-lactamase inhibitors (7.9%), together, represented 41.1% of the remaining antibiotic prescriptions. The choice of the route of administration was significantly influenced by age of the patients, by symptoms and signs of the disease and by the presence of concurrent diseases rather than by bacteria suspected of causing the disease. The rather marked variation in antibiotic prescribing pattern for LRTIs among Sicilian GPs reflects lack of availability or knowledge of any local or national guidelines about the management of these diseases. PMID- 10378991 TI - Disruption of temporo-entorhinal connections abolishes the facilitatory effect of angiotensins on memory in rats. AB - It has been found in our laboratory that the positive influence of angiotensin II (AII) and its 3-7 fragment [AII(3-7)] on learning and memory processes is mediated by the excitatory amino acids, since it was abolished by NMDA receptor antagonists. The purpose of the present study was to investigate whether bilateral disruption of glutamatergic temporo- entorhinal connections may have an influence on the facilitatory effect of both angiotensin peptides on memory motivated affectively. The bilateral transections of temporo-entorhinal connections were made in 27 male rats 10 days before testing the influence of intracerebroventricular AII and AII(3-7) injection on retrieval of a passive avoidance response. Twenty-seven additional rats served as sham-operated controls. Twenty-five lesioned and 25 sham-operated animals were accepted to the final analysis. AII and its 3-7 fragment significantly improved the retrieval process in sham-operated groups of rats. Bilateral disruption of temporo entorhinal connections totally abolished the facilitatory effect of both angiotensins on recall of information in a passive avoidance situation. Moreover, bilateral disruption of temporo-entorhinal connections markedly but not significantly attenuated crossings of squares, evaluated in an open field test, without an influence on rearings and bar approaches. These results may suggest that in the facilitatory effect of AII and AII(3-7) on memory motivated affectively involves reciprocal glutamatergic connection between lateral entorhinal cortex and temporal cortex. PMID- 10378992 TI - Inhibitory effect of mast cell-mediated immediate-type allergic reactions by Cichorium intybus. AB - We investigated the effect of an aqueous extract of Cichorium intybus (CIAE) on mast cell-mediated immediate type allergic reactions. CIAE (0.1-1000 mg kg-1) dose-dependently inhibited systemic anaphylactic reaction induced by compound 48/80 in mice. Especially, CIAE inhibited compound 48/80-induced anaphylactic reaction 100% with the dose of 1000 mg kg-1. CIAE 1000 mg kg-1also significantly inhibited local anaphylactic reaction activated by anti-dinitrophenyl (DNP) IgE. When mice were pretreated with CIAE at a concentration ranging from 0.1 to 1000 mg kg-1, the plasma histamine levels were reduced in a dose-dependent manner. CIAE (1-1000 microg ml-1) dose-dependently inhibited histamine release from the rat peritoneal mast cells (RPMC) activated by compound 48/80 or anti-DNP IgE. The level of cAMP in RPMC, when CIAE (1000 microg ml-1) was added, increased significantly compared with that of control cells. These results indicate that CIAE inhibits mast cell-mediated immediate-type allergic reactions in vivo and in vitro. PMID- 10378993 TI - Pharmacokinetics of paclitaxel administered as a 3-hour or 96-hour infusion. AB - AIM: To investigate the pharmacokinetics of paclitaxel (Paxene) administered to patients with advanced breast or ovarian cancer and to document safety and anti tumour activity in this study population. PATIENTS AND METHODS: Patients with advanced breast or ovarian cancer were accrued to two clinical studies. Paclitaxel (Paxene) was administered as a 3-h 175 mg m-2 or as a 96-h 140 mg m 2(105 mg m-2 in the presence of liver metastases) infusion. Patients not responding to the 3-h schedule were permitted to cross-over to the 96-h schedule. The data were compared to those of five patients who were previously treated with paclitaxel administered as Taxol (140 mg m-296-h infusion) at our Institute. RESULTS: Fourteen patients with breast cancer and five ovarian cancer patients were entered into this study. Seven patients received the 3-h regimen, and 12 were assigned to the 96-h schedule. Five patients originally treated with the 3-h schedule, crossed over to the 96-h arm. For the 3-h 175 mg m-2 dose, the area under the plasma concentration vs time curve (AUC) was (mean+/-SD) 16.9+/-4.8 h x micromol x l-1, whereas the AUCs were 5.5+/-1.2 and 4.3+/-0.9 h x micromol x l-1 for the 96-h 140 mg m-2 and 105 mg m-2 doses, respectively. The clearance of paclitaxel was independent of the dose in the 96-h group, indicating linear pharmacokinetics. Pharmacokinetics of Paxene (96-h 140 mg m-2) were not significantly different from the kinetics after Taxol (96-h 140 mg m-2) administration. PMID- 10378994 TI - Effect of ulceration on rat gastric tissue polyamine contents in response to different procedures; inhibition of these effects by cimetidine. AB - The effects of cimetidine an H2 receptor histamine antagonist on aspirin- and cold-restraint-stress-induced gastric lesions have been studied in rats. Cimetidine had a pronounced inhibitory effect on gastric lesions induced by either oral administration of aspirin (400 mg kg-1) or by cold-restraint stress in rats. These inhibitory effects were dose-related in the aspirin treatment group being 47 and 85% (P<0.05 and P<0.001) at 37.5 and 75 mg kg-1 doses, respectively, when compared to the control. Cimetidine was found effective in cold stress but inhibition with the low dose was not significant. However, high dose (75 mg kg-1) showed a significant reduction (P<0.01) in lesion index. In another series of experiments with the same regimen, the effects of different ulcerogenic procedures on the rat gastric tissue polyamine contents (putrescine, spermine and spermidine) and monoacetyl derivatives (N1- and N8-acetylspermidine) have been investigated by using HPLC method. The procedure permits use of n octane sulphonate as an ion pairing agent on the reversed-phase column. The treatment of rats with aspirin caused a substantial decrease in the concentration of different polyamine contents in the glandular part of stomach tissue. Pretreatment with cimetidine showed a marked protection against this decline in polyamine contents at both the doses tested (37.5 mg kg-1 and 75 mg kg-1) and increased the contents of spermidine and spermine above the control values. In the other part, cold-restraint stress also declined the polyamine contents. Low dose of cimetidine was found ineffective in this model. However, a high dose of cimetidine caused a significant rise in the levels of spermidine and spermine (P<0.001 and P<0.01, respectively) above the control levels. These findings suggest that cimetidine, besides being a H2-receptor antagonist, prevents ulcer formation due to its growth promotional properties, possibly through an increase in tissue polyamine contents that offer a defense barrier against the oxygen derived free radicals involved in the etiology of ulceration. It is also suggested that the rise in polyamine contents of gastric tissue is a crucial event in cytoprotection against destructive stimuli. PMID- 10378995 TI - The cost-effectiveness of low-molecular-weight heparin vs unfractionated heparin in general and orthopaedic surgery: an analysis for the German healthcare system. AB - BACKGROUND: Studies over the last years provided empirical evidence that low molecular weight heparins (LMWH) have advantages over unfractionated heparin (UFH) as post-operative thromboprophylactic agents. A detailed perspective economic evaluation for Germany is not available. METHODS: A deterministic decision-tree analysis was developed to estimate the main cost-relevant pathophysiological events: deep vein thrombosis (DTV), pulmonary embolism (PE) and major bleeding complications (BC) in general and orthopaedic surgery. The specific costs were analysed, summed up and put into relation with saved lifeyears from the perspective of society, third party payer and hospital management. The robustness of these results were substantiated by a detailed sensitivity analysis. RESULT: LMWH avoids pathophysiological events, saves costs and lifeyears in general and orthopaedic surgery from all perspectives. CONCLUSION: The use of LMWH as post-operative thromboprophylactic agents is more cost-effective than UFH in Germany. PMID- 10378996 TI - Environmental exposure to volatile organic compounds among workers in Mexico City as assessed by personal monitors and blood concentrations. AB - Benzene, an important component in gasoline, is a widely distributed environmental contaminant that has been linked to known health effects in animals and humans, including leukemia. In Mexico City, environmental benzene levels, which may be elevated because of the heavy traffic and the poor emission control devices of older vehicles, may pose a health risk to the population. To assess the potential risk, portable passive monitors and blood concentrations were used to survey three different occupational groups in Mexico City. Passive monitors measured the personal exposure of 45 workers to benzene, ethylbenzene, toluene, o xylene and m-/p-xylene during a work shift. Blood concentrations of the above volatile organic compounds (VOCs), methyl tert-butyl ether, and styrene were measured at the beginning and the end of a work shift. Passive monitors showed significantly higher (p > 0.0001) benzene exposure levels among service station attendants (median = 330 microg/m3; range 130-770) as compared to street vendors (median = 62 microg/m3; range 49-180) and office workers (median = 44 microg/m3, range 32-67). Baseline blood benzene levels (BBLs) for these groups were higher than those reported for similar populations from Western countries (median = 0.63 microg/L, n = 24 for service station attendants; median = 0.30 microg/L, n = 6 for street vendors; and median = 0.17 microgr;g/L, n = 7 for office workers). Nonsmoking office workers who were nonoccupationally exposed to VOCs had BBLs that were more than five times higher than those observed in a nonsmoking U.S. population. BBLs of participants did not increase during the work shift, suggesting that because the participants were chronically exposed to benzene, complex pharmacokinetic mechanisms were involved. Our results highlight the need for more complete studies to assess the potential benefits of setting environmental standards for benzene and other VOCs in Mexico. PMID- 10378997 TI - Collection of genomic DNA by buccal swabs for polymerase chain reaction-based biomarker assays. AB - Studies in molecular and genetic epidemiology require a high-throughput, low cost, and reliable means of genomic DNA collection. Buccal (cheek) swabs have been proposed as a means of achieving these goals, but there is little information about the practical application of this approach. From January 1995 to December 1997, we processed 995 buccal swabs for use in polymerase chain reaction (PCR)-based genotype assays in the context of ongoing molecular epidemiologic studies. Six hundred forty-seven of these swabs were processed immediately after collection and 348 were received by mail. We were able to obtain at least one genotype from 99.7% (645 of 647) of fresh-processed and 97.4% (330 of 339) of mailed biosamples. A PCR success rate of 90.3% (2,546 genotypes from 2,819 assays) was achieved. Genotypes were obtained from 96.1% (1, 865 genotypes from 1,941 assays) of fresh-processed biosamples and 77.6% (681 genotypes from 878 assays) of mailed biosamples. PCR success rates at any single locus ranged from 92.6 to 98.8% (fresh-processed) and 75.5 to 79.6% (mailed). The PCR success rate among fresh-processed biosamples was significantly higher than among mailed biosamples (Fisher's exact test p < 0.0001), and more attempts were required to obtain a successful PCR result for mailed biosamples as compared to fresh-processed biosamples. For one locus (CYP3A4), a subset of mailed biosamples was purified if two or more PCR failures occurred. Additional genotypes were obtained in 58.3% of these previously failed biosamples. Time from biosample receipt to DNA extraction had no effect on PCR success. After storage of processed biosamples for as long as 3 years, there was no appreciable decrease in the rate of PCR success. These results suggest that adequate DNA for PCR-based applications can be obtained from buccal swabs, but sampling or processing considerations may be important in obtaining optimal results. PMID- 10378998 TI - Daily variation of particulate air pollution and poor cardiac autonomic control in the elderly. AB - examined the cardiac autonomic response to daily variations in PM in 26 elderly (mean age 81) individuals for 3 consecutive weeks. Several standardized methods were used to measure 24-hr average PM concentrations prior to the clinical test inside (indoor PM2.5) and immediately outside (outdoor PM2.5 and PM2.5-10) of participants' residences. Resting, supine, 6-min R wave to R wave (R-R) interval data were collected to estimate high frequency (0.15-0.40 Hz) and low frequency (0.04-0.15 Hz) powers and standard deviation of normal R-R intervals (SDNN) as cardiac autonomic control indices. Participant-specific lower heart rate variability days were defined as days for which the high-frequency indices fell below the first tertile of the individual's high-frequency distribution over the study period. Indoor PM2.5 > 15 microg/m3 was used to define high pollution days. Results show that the odds ratio (95% confidence interval) of low heart rate variability high frequency for high (vs. not high) pollution days was 3.08 (1.43, 6.59). The ss-coefficients (standard error) from mixed models to assess the quantitative relationship between variations in indoor PM2.5 and the log transformed high frequency, low frequency, and SDNN were: -0.029 (0.010), -0.027 (0.009), and -0.004 (0.003), respectively. This first study of cardiac autonomic control response to daily variations of PM2.5 indicates that increased levels of PM2.5 are associated with lower cardiac autonomic control, suggesting a possible mechanistic link between PM and cardiovascular disease mortality. PMID- 10378999 TI - Radionuclides in the lichen-caribou-human food chain near uranium mining operations in northern Saskatchewan, Canada. AB - The richest uranium ore bodies ever discovered (Cigar Lake and McArthur River) are presently under development in northeastern Saskatchewan. This subarctic region is also home to several operating uranium mines and aboriginal communities, partly dependent upon caribou for subsistence. Because of concerns over mining impacts and the efficient transfer of airborne radionuclides through the lichen-caribou-human food chain, radionuclides were analyzed in tissues from 18 barren-ground caribou (Rangifer tarandus groenlandicus). Radionuclides included uranium (U), radium (226Ra), lead (210Pb), and polonium (210Po) from the uranium decay series; the fission product (137Cs) from fallout; and naturally occurring potassium (40K). Natural background radiation doses average 2-4 mSv/year from cosmic rays, external gamma rays, radon inhalation, and ingestion of food items. The ingestion of 210Po and 137Cs when caribou are consumed adds to these background doses. The dose increment was 0.85 mSv/year for adults who consumed 100 g of caribou meat per day and up to 1.7 mSv/year if one liver and 10 kidneys per year were also consumed. We discuss the cancer risk from these doses. Concentration ratios (CRs), relating caribou tissues to lichens or rumen (stomach) contents, were calculated to estimate food chain transfer. The CRs for caribou muscle ranged from 1 to 16% for U, 6 to 25% for 226Ra, 1 to 2% for 210Pb, 6 to 26% for 210Po, 260 to 370% for 137Cs, and 76 to 130% for 40K, with 137Cs biomagnifying by a factor of 3-4. These CRs are useful in predicting caribou meat concentrations from the lichens, measured in monitoring programs, for the future evaluation of uranium mining impacts on this critical food chain. PMID- 10379000 TI - Inhalation of diesel engine exhaust affects spermatogenesis in growing male rats. AB - We conducted experiments to determine whether diesel engine exhaust affects reproductive endocrine function in growing rats. The rats were assigned to three groups: a group exposed to total diesel engine exhaust containing 5.63 mg/m3 particulate matter, 4.10 ppm nitrogen dioxide, and 8.10 ppm nitrogen oxide; a group exposed to filtered exhaust without particulate matter; and a group exposed to clean air. Dosing experiments were performed for 3 months beginning at birth (6 hr/day for 5 days/week). Serum levels of testosterone and estradiol were significantly higher in animals exposed to total diesel exhaust and filtered exhaust (p < 0.05 for each group) as compared to the controls. Follicle stimulating hormone was significantly decreased in the two groups exposed to diesel exhaust as compared to the control group (p < 0.05). Luteinizing hormone was significantly decreased in the total exhaust-exposed group as compared to the control and filtered groups (p < 0.05). Although testis weight did not show any significant difference among the groups, sperm production and activity of testicular hyaluronidase were significantly reduced in both exhaust-exposed groups as compared to the control group. Histological examination showed decreased numbers of step 18 and 19 spermatids in stage VI, VII, and VIII tubules in the testes of both diesel exhaust-exposed groups. This study suggests that diesel exhaust stimulates hormonal secretion of the adrenal cortex, depresses gonadotropin-releasing-hormone, and inhibits spermatogenesis in rats. Because these effects were not inhibited by filtration, the gaseous phase of the exhaust appears to be more responsible than particulate matter for disrupting the endocrine system. PMID- 10379001 TI - Heat shock protein 27 expression in human proximal tubule cells exposed to lethal and sublethal concentrations of CdCl2. AB - The expression of hsp 27 mRNA and protein was determined in cultured human proximal tubule (HPT) cells exposed to lethal and sublethal concentrations of Cd2+ under both acute and extended conditions. Initial procedures demonstrated that HPT cells display the classic stress response following physical and chemical stress. Heat stress (42.5 degrees C for 1 hr) caused an increase in both hsp 27 mRNA and protein as well as a shift in the protein to a more phosphorylated state. Results were similar when the cells were subjected to chemical stress (exposure to 100 microM sodium arsenite for 4 hr). Acute exposure to 53 microM CdCl2 for 4 hr also resulted in an increase in hsp 27 mRNA and protein and a shift to the more phosphorylated protein isoform. Extended Cd2+ exposure involved continuous treatment with Cd2+ at both lethal and sublethal levels over a 16-day time course. The results of this treatment showed that chronic exposure to Cd2+ failed to increase either hsp 27 mRNA or protein expression in HPT cells, even at lethal Cd2+ concentrations. In fact, hsp 27 protein levels decreased as compared to controls at both lethal and sub-lethal exposure to Cd2+. These findings imply that hsp 27 expression in human proximal tubule cells may have two distinct modes depending on the nature (acute vs. chronic) of the stress. PMID- 10379002 TI - An assay for the detection of xenoestrogens based on a promoter containing overlapping EREs. AB - Xenoestrogens could be implicated in the decrease of male fertility and in the increased incidence of testicular and breast cancers in humans. To predict their deleterious effects, various in vivo or in vitro tests have been proposed to assay the xenoestrogenic activity. We have designed an assay for the detection of xenoestrogens based on a novel estrogen responsive unit formed by two overlapping estrogen response elements (overEREs). This construct is able to mediate a synergistic activation of transcription by 17ss-estradiol. We have used the overERE unit to assay the estrogenic activity of synthetic compounds, mostly organochlorine compounds. By using the overERE construct, we were able to detect the estrogenic activity of compounds at concentrations 10- to 100-fold lower than a single ERE (i.e., we detected the estrogenic effect of endosulfan at a concentration of 10(-5) M with ERE, whereas the overERE unit allowed us to detect a significant estrogenic activity of endosulfan at a lower concentration (10(-6) M). Some compounds did not exhibit any estrogenic activity when tested with a classical ERE, whereas they were potent xenoestrogens when the overERE was used (i.e., Betanal). The assays we have developed are very sensitive and can be performed quickly. Moreover, because the promoter that we used contains only an overlapping ERE as a regulatory unit, the interference of the tested molecules with other regulatory pathways can be avoided. PMID- 10379003 TI - Particulate air pollution and daily mortality on Utah's Wasatch Front. AB - Reviews of daily time-series mortality studies from many cities throughout the world suggest that daily mortality counts are associated with short-term changes in particulate matter (PM) air pollution. One U.S. city, however, with conspicuously weak PM-mortality associations was Salt Lake City, Utah; however, relatively robust PM-mortality associations have been observed in a neighboring metropolitan area (Provo/Orem, Utah). The present study explored this apparent discrepancy by collecting, comparing, and analyzing mortality, pollution, and weather data for all three metropolitan areas on Utah's Wasatch Front region of the Wasatch Mountain Range (Ogden, Salt Lake City, and Provo/Orem) for approximately 10 years (1985-1995). Generalized additive Poisson regression models were used to estimate PM-mortality associations while controlling for seasonality, temperature, humidity, and barometric pressure. Salt Lake City experienced substantially more episodes of high PM that were dominated by windblown dust. When the data were screened to exclude obvious windblown dust episodes and when PM data from multiple monitors were used to construct an estimate of mean exposure for the area, comparable PM-mortality effects were estimated. After screening and by using constructed mean PM [less than/equal to] 10 microm in aerodynamic diameter (PM10) data, the estimated percent change in mortality associated with a 10-mg/m3 increase in PM10 (and 95% confidence intervals) for the three Wasatch Front metropolitan areas equaled approximately 1. 6% (0.3-2.9), 0.8% (0.3-1.3), and 1.0% (0.2-1.8) for the Ogden, Salt Lake City, and Provo/Orem areas, respectively. We conclude that stagnant air pollution episodes with higher concentrations of primary and secondary combustion-source particles were more associated with elevated mortality than windblown dust episodes with relatively higher concentrations of coarse crustal-derived particles. PMID- 10379004 TI - Immune responses in farm workers after exposure to Bacillus thuringiensis pesticides. AB - Although health risks to pesticides containing Bacillus thuringiensis (Bt) have been minimal, the potential allergenicity of these organisms has not been evaluated. Therefore, a health survey was conducted in farm workers before and after exposure to Bt pesticides. Farm workers who picked vegetables that required Bt pesticide spraying were evaluated before the initial spraying operation (n = 48) and 1 and 4 months after (n = 32 and 20, respectively). Two groups of low- (n = 44) and medium- (n = 34) exposure workers not directly exposed to Bt spraying were also assessed. The investigation included questionnaires, nasal/mouth lavages, ventilatory function assessment, and skin tests to indigenous aeroallergens and to a variety of Bt spore and vegetative preparations. To authenticate exposure to the organism present in the commercial preparation, isolates from lavage specimens were tested for Bt genes by DNA-DNA hybridization. Humoral immunoglobulin G (IgG) and immunoglobulin E (IgE) antibody responses to spore and vegetative Bt extracts were assayed. There was no evidence of occupationally related respiratory symptoms. Positive skin-prick tests to several spore extracts were seen chiefly in exposed workers. In particular, there was a significant (p < 0.05) increase in the number of positive skin tests to spore extracts 1 and 4 months after exposure to Bt spray. The number of positive skin test responses was also significantly higher in high (p < 0.05) than in low- or medium-exposure workers. The majority of nasal lavage cultures from exposed workers was positive for the commercial Bt organism, as demonstrated by specific molecular genetic probes. Specific IgE antibodies were present in more high exposure workers (p < 0.05) than in the low and medium groups. Specific IgG antibodies occurred more in the high (p < 0.05) than in the low-exposure group. Specific IgG and IgE antibodies to vegetative organisms were present in all groups of workers. Exposure to Bt sprays may lead to allergic skin sensitization and induction of IgE and IgG antibodies, or both. PMID- 10379005 TI - Infantile methemoglobinemia: reexamining the role of drinking water nitrates. AB - Ingestion of nitrates in drinking water has long been thought to be a primary cause of acquired infantile methemoglobinemia, often called blue baby syndrome. However, recent research and a review of historical cases offer a more complex picture of the causes of infantile methemoglobinemia. Gastrointestinal infection and inflammation and the ensuing overproduction of nitric oxide may be the cause of many cases of infantile methemoglobinemia previously attributed to drinking water nitrates. If so, current limits on allowable levels of nitrates in drinking water, which are based solely on the health threat of infantile methemoglobinemia, may be unnecessarily strict. PMID- 10379006 TI - Methylmercury neurotoxicity in Amazonian children downstream from gold mining. AB - In widespread informal gold mining in the Amazon Basin, mercury is used to capture the gold particles as amalgam. Releases of mercury to the environment have resulted in the contamination of freshwater fish with methylmercury. In four comparable Amazonian communities, we examined 351 of 420 eligible children between 7 and 12 years of age. In three Tapajos villages with the highest exposures, more than 80% of 246 children had hair-mercury concentrations above 10 microg/g, a limit above which adverse effects on brain development are likely to occur. Neuropsychological tests of motor function, attention, and visuospatial performance showed decrements associated with the hair-mercury concentrations. Especially on the Santa Ana form board and the Stanford-Binet copying tests, similar associations were also apparent in the 105 children from the village with the lowest exposures, where all but two children had hair-mercury concentrations below 10 microg/g. Although average exposure levels may not have changed during recent years, prenatal exposure levels are unknown, and exact dose relationships cannot be generated from this cross-sectional study. However, the current mercury pollution seems sufficiently severe to cause adverse effects on brain development. PMID- 10379007 TI - Arsenic: health effects, mechanisms of actions, and research issues. AB - A meeting on the health effects of arsenic (As), its modes of action, and areas in need of future research was held in Hunt Valley, Maryland, on 22-24 September 1997. Exposure to As in drinking water has been associated with the development of skin and internal cancers and noncarcinogenic effects such as diabetes, peripheral neuropathy, and cardiovascular diseases. There is little data on specific mechanism(s) of action for As, but a great deal of information on possible modes of action. Although arsenite [As(III)] can inhibit more than 200 enzymes, events underlying the induction of the noncarcinogenic effects of As are not understood. With respect to carcinogenicity, As can affect DNA repair, methylation of DNA, and increase radical formation and activation of the protooncogene c-myc, but none of these potential pathways have widespread acceptance as the principal etiologic event. In addition, there are no accepted models for the study of As-induced carcinogenesis. At the final meeting session we considered research needs. Among the most important areas cited were a) As metabolism and its interaction with cellular constituents; b) possible bioaccumulation of As; c) interactions with other metals; d) effects of As on genetic material; e) development of animal models and cell systems to study effects of As; and f) a better characterization of human exposures as related to health risks. Some of the barriers to the advancement of As research included an apparent lack of interest in the United States on As research; lack of relevant animal models; difficulty with adoption of uniform methodologies; lack of accepted biomarkers; and the need for a central storage repository for stored specimens. PMID- 10379008 TI - Occupational asthma and contact dermatitis in a spray painter after introduction of an aziridine cross-linker. AB - A 23-year-old spray painter developed contact dermatitis and respiratory difficulty characterized by small airways obstruction shortly after the polyfunctional aziridine cross-linker CX-100 began to be used in his workplace as a paint activator. The symptoms resolved after he was removed from the workplace and was treated with inhaled and topical steroids. Painters may have an increased risk of asthma due to exposure to a variety of agents, such as isocyanates, alkyd resins, and chromates. This case illustrates the importance of using appropriate work practices and personal protective equipment to minimize exposure. Occupational asthma is diagnosed by a history of work-related symptoms and exposure to known causative agents. The diagnosis is confirmed by serial pulmonary function testing or inhalational challenge testing. The risk of asthma attributable to occupational exposures is probably underappreciated due to underreporting and to inappropriate use of narrow definitions of exposure in epidemiologic studies of attributable risk. PMID- 10379010 TI - Xenobiotics and cell Death/Injury in neurodegenerative diseases AB - With the "graying of America," the prevalence of neurodegenerative diseases such as Alzheimer disease (AD) and Parkinson disease (PD) will make increasingly greater demands on Medicare/Medicaid resources, which will greatly strain the national economy. It is imperative that prevention and intervention strategies for these diseases be developed now in order to prevent a bleak future impact. PMID- 10379009 TI - An unusual case of carbon monoxide poisoning. AB - Carbon monoxide, a gas originating from incomplete combustion of carbon-based fuels, is an important cause of human deaths. In this paper, we describe an unusual carbon monoxide poisoning in a dwelling without obvious sources of combustion gases, for which two adults had to be treated in a hyperbaric chamber. Carbon monoxide readings were taken in the house and in the neighboring homes. Methane gas and nitrogen oxide levels were also monitored in the house air. Soil samples were collected around the house and tested for hydrocarbon residues. The investigation revealed the presence of a pocket of carbon monoxide under the foundation of the house. The first readings revealed carbon monoxide levels of 500 ppm in the basement. The contamination lasted for a week. The investigation indicated that the probable source of contamination was the use of explosives at a nearby rain sewer construction site. The use of explosives in a residential area can constitute a major source of carbon monoxide for the neighboring populations. This must be investigated, and public health authorities, primary care physicians, governmental authorities, and users and manufacturers of explosives must be made aware of this problem. PMID- 10379011 TI - A holistic approach to environmental health research. AB - One of the most formidable questions facing the environmental health research community today is how to translate basic fundamental research into a product (e.g., disease outcome) that meets the needs of its stakeholders--the medical community, regulatory agencies, and ultimately, the citizens of our nation. Historically, a successful research program could be defined as one that received continuous funding, produced high-quality publications, and was respected by scientists in related fields. However, it is now apparent that this is not sufficient for attaining improved public health--the ultimate goal of these research efforts. Research results must be transferred in a more active way to the communities and professionals who have need of this information. The link must be recognized, and the roles of the stakeholders in the entire research process must be acknowledged to ensure full impact of the research endeavors. PMID- 10379012 TI - A world-class program. AB - It is said that if you give a man a fish you feed him for a day, but if you teach him to fish you feed him for a lifetime. In the spirit of this adage, the International Training and Research in Environmental and Occupational Health (ITREOH) program trains health scientists, clinicians, epidemiologists, toxicologists, engineers, industrial hygienists, chemists, and allied health workers from developing countries in the skills they need to maintain environmental and occupational health in their countries both now and in the future. PMID- 10379013 TI - A healthy home environment? AB - Over the past seven years, the U.S. Environmental Protection Agency has consistently ranked indoor air pollution among the top five risks to public health. One of the most dangerous indoor air pollutants is carbon monoxide (CO). CO can be lethal, but perhaps more important, many people suffer ill health from chronic, often undetected exposure to low levels of this gas, resulting in fatigue, headache, dizziness, nausea, and vomiting. Another dangerous pollutant is volatile organic compounds (VOCs), which come from sources including building products, cleaning agents, and paints. One VOC, formaldehyde, can act as an irritant to the conjunctiva and upper and lower respiratory tract. Formaldehyde is also known to cause nasal cancer in test animals. PMID- 10379014 TI - Chloroform: An EPA test case. AB - In March 1998, the U.S. Environmental Protection Agency (EPA) published a proposal to raise the drinking water maximum contaminant level goal (MCLG) for chloroform, a suspected human carcinogen, from zero to 300 parts per billion. The proposal marked a departure from the agency's traditional reliance on linear dose response models in performing risk assessment, and reflected the new thinking contained in the 1996 draft update to the agency's cancer risk assessment guidelines. The updated guidelines emphasize mechanisms of action and descriptions of the conditions under which carcinogenic hazards are likely to be expressed. PMID- 10379015 TI - Floorward thinking. AB - From its creation to its disposal, there are environmental and health problems associated with polyvinyl chloride (vinyl), the major component of vinyl flooring. The production of vinyl creates toxic waste that must be dumped or incinerated. Because very little vinyl is recycled, the waste material must also be landfilled or burned. Furthermore, the heavy chlorine content of these materials result in the release of dioxins into the environment. A new flooring alternative to vinyl recently entered the market. Stratica, manufactured by Amtico Company Limited based in Coventry, United Kingdom, is made from polymer resins and offers the durability of vinyl without the environmental impact. PMID- 10379016 TI - Effects of different forms of dietary hydrogenated fats on serum lipoprotein cholesterol levels. AB - BACKGROUND: Metabolic studies suggest that fatty acids containing at least one double bond in the trans configuration, which are found in hydrogenated fat, have a detrimental effect on serum lipoprotein cholesterol levels as compared with unsaturated fatty acids containing double bonds only in the cis configuration. We compared the effects of diets with a broad range of trans fatty acids on serum lipoprotein cholesterol levels. METHODS: Eighteen women and 18 men consumed each of six diets in random order for 35-day periods. The foods were identical in each diet, and each diet provided 30 percent of calories as fat, with two thirds of the fat contributed as soybean oil (<0.5 g of trans fatty acid per 100 g of fat), semiliquid margarine (<0.5 g per 100 g), soft margarine (7.4 g per 100 g), shortening (9.9 g per 100 g), or stick margarine (20.1 g per 100 g). The effects of those diets on serum lipoprotein cholesterol, triglyceride, and apolipoprotein levels were compared with those of a diet enriched with butter, which has a high content of saturated fat. RESULTS: The mean (+/-SD) serum low-density lipoprotein (LDL) cholesterol level was 177+/-32 mg per deciliter (4.58+/-0.85 mmol per liter) and the mean high-density lipoprotein (HDL) cholesterol level was 45+/-10 mg per deciliter (1.2+/-0.26 mmol per liter) after subjects consumed the butter enriched diet. The LDL cholesterol level was reduced on average by 12 percent, 11 percent, 9 percent, 7 percent, and 5 percent, respectively, after subjects consumed the diets enriched with soybean oil, semiliquid margarine, soft margarine, shortening, and stick margarine; the HDL cholesterol level was reduced by 3 percent, 4 percent, 4 percent, 4 percent, and 6 percent, respectively. Ratios of total cholesterol to HDL cholesterol were lowest after the consumption of the soybean-oil diet and semiliquid-margarine diet and highest after the stick margarine diet. CONCLUSIONS: Our findings indicate that the consumption of products that are low in trans fatty acids and saturated fat has beneficial effects on serum lipoprotein cholesterol levels. PMID- 10379017 TI - Effect of systemic glucocorticoids on exacerbations of chronic obstructive pulmonary disease. Department of Veterans Affairs Cooperative Study Group. AB - BACKGROUND AND METHODS: Although their clinical efficacy is unclear and they may cause serious adverse effects, systemic glucocorticoids are a standard treatment for patients hospitalized with exacerbations of chronic obstructive pulmonary disease (COPD). We conducted a double-blind, randomized trial of systemic glucocorticoids (given for two or eight weeks) or placebo in addition to other therapies, for exacerbations of COPD. Most other care was standardized over the six-month period of follow-up. The primary end point was treatment failure, defined as death from any cause or the need for intubation and mechanical ventilation, readmission to the hospital for COPD, or intensification of drug therapy. RESULTS: Of 1840 potential study participants at 25 Veterans Affairs medical centers, 271 were eligible for participation and were enrolled; 80 received an eight-week course of glucocorticoid therapy, 80 received a two-week course, and 111 received placebo. About half the potential participants were ineligible because they had received systemic glucocorticoids in the previous 30 days. Rates of treatment failure were significantly higher in the placebo group than in the two glucocorticoid groups combined at 30 days (33 percent vs. 23 percent, P=0.04) and at 90 days (48 percent vs. 37 percent, P=0.04). Systemic glucocorticoids (in both groups combined) were associated with a shorter initial hospital stay (8.5 days, vs. 9.7 days for placebo, P=0.03) and with a forced expiratory volume in one second that was about 0.10 liter higher than that in the placebo group by the first day after enrollment. Significant treatment benefits were no longer evident at six months. The eight-week regimen of therapy was not superior to the two-week regimen. The patients who received glucocorticoid therapy were more likely to have hyperglycemia requiring therapy than those who received placebo (15 percent vs. 4 percent, P=0.002). CONCLUSIONS: Treatment with systemic glucocorticoids results in moderate improvement in clinical outcomes among patients hospitalized for exacerbations of COPD. The maximal benefit is obtained during the first two weeks of therapy. Hyperglycemia of sufficient severity to warrant treatment is the most frequent complication. PMID- 10379018 TI - Long-term treatment with inhaled budesonide in persons with mild chronic obstructive pulmonary disease who continue smoking. European Respiratory Society Study on Chronic Obstructive Pulmonary Disease. AB - BACKGROUND: Although patients with chronic obstructive pulmonary disease (COPD) should stop smoking, some do not. In a double-blind, placebo-controlled study, we evaluated the effect of the inhaled glucocorticoid budesonide in patients with mild COPD who continued smoking. After a six-month run-in period, we randomly assigned 1277 subjects (mean age, 52 years; mean forced expiratory volume in one second [FEV1], 77 percent of the predicted value; 73 percent men) to twice-daily treatment with 400 microg of budesonide or placebo, inhaled from a dry-powder inhaler, for three years. RESULTS: Of the 1277 subjects, 912 (71 percent) completed the study. Among these subjects, the median decline in the FEV1 after the use of a bronchodilator over the three-year period was 140 ml in the budesonide group and 180 ml in the placebo group (P=0.05), or 4.3 percent and 5.3 percent of the predicted value, respectively. During the first six months of the study, the FEV1 improved at the rate of 17 ml per year in the budesonide group, as compared with a decline of 81 ml per year in the placebo group (P<0.001). From nine months to the end of treatment, the FEV1 declined at similar rates in the two groups (P=0.39). Ten percent of the subjects in the budesonide group and 4 percent of those in the placebo group had skin bruising (P<0.001). Newly diagnosed hypertension, bone fractures, postcapsular cataracts, myopathy, and diabetes occurred in less than 5 percent of the subjects, and the diagnoses were equally distributed between the groups. CONCLUSIONS: In patients with mild COPD who continue smoking, the use of inhaled budesonide is associated with a small one-time improvement in lung function but does not appreciably affect the long term progressive decline. PMID- 10379019 TI - Gain of chromosome arm 17q and adverse outcome in patients with neuroblastoma. AB - BACKGROUND: Gain of genetic material from chromosome arm 17q (gain of segment 17q21-qter) is the most frequent cytogenetic abnormality of neuroblastoma cells. This gain has been associated with advanced disease, patients who are > or =1 year old, deletion of chromosome arm 1p, and amplification of the N-myc oncogene, all of which predict an adverse outcome. We investigated these associations and evaluated the prognostic importance of the status of chromosome 17. METHODS: We compiled molecular cytogenetic analyses of chromosome 17 in primary neuroblastomas in 313 patients at six European centers. Clinical and survival information were collected, along with data on 1p, N-myc, and ploidy. RESULTS: Unbalanced gain of segment 17q21-qter was found in 53.7 percent of the tumors, whereas the chromosome was normal in 46.3 percent. The gain of 17q was characteristic of advanced tumors and of tumors in children > or =1 year of age and was strongly associated with the deletion of 1p and amplification of N-myc. No tumor showed amplification of N-myc in the absence of either deletion of 1p or gain of 17q. Gain of 17q was a significant predictive factor for adverse outcome in univariate analysis. Among the patients with this abnormality, overall survival at five years was 30.6 percent (95 percent confidence interval, 21 to 40 percent), as compared with 86.0 percent (95 percent confidence interval, 78 to 91 percent) among those with normal 17q status. in multivariate analysis, gain of 17q was the most powerful prognostic factor, followed by the presence of stage 4 disease and deletion of 1p (hazard ratios, 3.4, 2.3, and 1.9, respectively). CONCLUSIONS: Gain of chromosome segment 17q21-qter is an important prognostic factor in children with neuroblastoma. PMID- 10379020 TI - Vitamin A supplementation for extremely-low-birth-weight infants. National Institute of Child Health and Human Development Neonatal Research Network. AB - BACKGROUND: Vitamin A supplementation may reduce the risk of chronic lung disease and sepsis in extremely-low-birth-weight infants. The results of our pilot study suggested that a dose of 5000 IU administered intramuscularly three times per week for four weeks was more effective than the lower doses given in past trials. METHODS: We performed a multicenter, blinded, randomized trial to assess the effectiveness and safety of this regimen as compared with sham treatment in 807 infants in need of respiratory support 24 hours after birth. The mean birth weight was 770 g in the vitamin A group and 769 g in the control group, and the respective gestational ages were 26.8 and 26.7 weeks. RESULTS: By 36 weeks' postmenstrual age, 59 of the 405 infants (15 percent) in the vitamin A group and 55 of the 402 infants (14 percent) in the control group had died. The primary outcome - death or chronic lung disease at 36 weeks' postmenstrual age - occurred in significantly fewer infants in the vitamin A group than in the control group (55 percent vs. 62 percent; relative risk, 0.89; 95 percent confidence interval, 0.80 to 0.99). Overall, 1 additional infant survived without chronic lung disease for every 14 to 15 infants who received vitamin A supplements. The proportions of infants in the vitamin A group and the control group who had signs of potential vitamin A toxicity were similar. The proportion of infants with serum retinol values below 20 microg per deciliter (0.70 micromol per liter) was lower in the vitamin A group than in the control group (25 percent vs. 54 percent, P<0.001). CONCLUSIONS: Intramuscular administration of 5000 IU of vitamin A three times per week for four weeks reduced biochemical evidence of vitamin A deficiency and slightly decreased the risk of chronic lung disease in extremely-low-birth-weight infants. PMID- 10379021 TI - Images in clinical medicine. Lipemia retinalis. PMID- 10379022 TI - Molecular basis of the neurodegenerative disorders. PMID- 10379023 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 19-1999. A 55-year-old man with a destructive bone lesion 17 months after liver transplantation. PMID- 10379024 TI - Glucocorticoid therapy for chronic obstructive pulmonary disease. PMID- 10379025 TI - Predicting outcome in neuroblastoma. PMID- 10379026 TI - Trans fatty acids and coronary heart disease. PMID- 10379028 TI - Vertical column hydroclassification of metal-contaminated soils. AB - The purpose of this work was to reduce soil volumes requiring aggressive treatment. A second purpose was to determine differences in separation due to distinct forms of the metal contamination and soil texture. The objectives were to apply hydroclassification and find mass and metal-contaminant distribution of four soils contaminated with heavy metals from firing ranges, a small arms incinerator, and an electroplating operation. The soils were slurried in water, sieved, and exposed to upward flowing water to separate the soil particles into four nominal size ranges. The popping furnace soil exhibited substantial lead among all particle size fractions. The firing range soils exhibited bimodal distributions. The electroplating soil exhibited a strong concentration of metals toward the <63 microm fraction. Attrition scrubbing moderately improved the enrichment of metals in several fractions. Extraction revealed the lead and chromium in the electroplating soil to be relatively immobile. These results suggest metal distributions are influenced by the different mechanisms of introduction into the soil. They also help to predict performance of processing options such as sieving hydroclassification and attrition scrubbing. PMID- 10379027 TI - Extraction of TNT from aggregate soil fractions. AB - Past explosives manufacture, disposal, and training activities have contaminated soil at many military facilities, posing health and environmental risks through contact, potential detonation, and leaching into ground water. While methods have been confirmed for extraction and measuring explosives concentration in soil, no work has addressed aggregate size material (the >2 mm gravel and cobbles) that often occurs with the smaller soil fractions. This paper describes methods and results for extraction and measurement of TNT (2,4,6-trinitrotoluene) in aggregate material from 1/2 to 2-1/1 from a WWII era ammunition plant. TNT was extracted into acetonitrile by both Soxhlet and ultrasonic extraction methods. High pressure liquid chromatography analyses of extracts showed expected variation among samples. Also effective extraction and determination of TNT concentration for each aggregate size fraction was achieved. PMID- 10379029 TI - Simplified soil washing processes for a variety of soils. AB - A soil washing process is described in which high shear mixing, sprays, hydrosizing, flotation, and screening are integrated to countercurrently clean sand, silt and clay. Chemical techniques are summarized that can be used to remove selected contaminants from the soil and then clean the wash water for reuse. Descriptions of simple and complex processes, and a recent project are used to illustrate the points: (1) Soil washing is not a single process, but a collection of unit operations assembled for each project; (2) all process must be coordinated for the project to be successful; and (3) combining techniques in a few pieces of equipment debottlenecks soil washing. PMID- 10379030 TI - Application of remedy studies to the development of a soil washing pilot plant that uses mineral processing technology: a practical experience. AB - Soil washing employing mineral processing technology to treat radionuclide contaminated soils has been examined as a remedy alternative to the exclusive excavation, transportation, and disposal of the soil. Successful application depends on a thorough remedy study, employing a systematic tiered approach that is efficient, self-limiting, and cost effective. The study includes: (1) site and soil characterization to determine the basic mineral and physical properties of both the soil and contaminants and to identify their relative associations; (2) treatment studies to evaluate the performance of process units for contaminant separation; (3) conceptual process design to develop a treatment pilot plant; and (4) engineering design to construct, test, and optimize the actual full-scale plant. A pilot plant using soil washing technology for the treatment of radium contaminated soil was developed, tested, and demonstrated. The plant used particle-size separation to produced a remediated product that represented approximately 50% of the contaminated soil. Subsequently, it was modified for more effective performance and application to soil with alternate characteristics; it awaits further testing. The economic analysis of soil washing using the pilot plant as a model indicates that a remedy plan based on mineral processing technology is very competitive with the traditional alternative employing excavation, transportation, and disposal exclusively, even when disposal costs are modest or when recovery of remediated soil during treatment is low. This paper reviews the tiered approach as it applies to mineral processing technology to treat radionuclide-contaminated soils and a pilot plant developed to test the soil washing process. PMID- 10379031 TI - Bench- and pilot-scale studies relating to the removal of uranium from uranium contaminated soils using carbonate and citrate lixiviants. AB - Development of the nuclear industry has resulted in soil becoming contaminated with uranium from a variety of sources. To avoid the disposal of these soils in conventional low-level radwaste burial sites, a technology is needed to extract/leach and concentrate uranium in soil into small volumes of an acceptable waste form and returning the soil to its original place. Two lixiviants, carbonate and citrate, were evaluated as to their ability to extract uranium from soil in a soil washing engineering process. The objective was to use a washing/extracting process to selectively remove the uranium from soil without seriously degrading the soil's physicochemical characteristics or generating a secondary waste form that is difficult to manage and/or dispose. Both carbonate and citric acid lixiviants were observed to be effective extractants to remove uranium from the soils tested. Carbonate, because of the its ability to be recycled and its tendency to be more selective for uranium, is preferred for most soils. A major obstacle for using citric acid as well as mineral-based acids is their generation of waste streams from which it is difficult to remove uranium and manage (and dispose of any residual waste water sludges) in an environmentally acceptable manner. The removal of uranium was examined for three soils sampled from two US Department of Energy sites. Two soils were from the facility formerly called the Feed Materials Production Center at Fernald, Ohio and the other soil was from the Oak Ridge Tennessee Y-12 Plant. In the bench scale studies, general relationships, such as the effect of carbonate and citrate concentrations, pH, the presence of oxidants, such as KMnO4, temperature, and extraction time were investigated. The best pilot-scale treatment consisted of three successive extractions with 0.25 M carbonate-bicarbonate (in presence of KMnO4 as an oxidant) at 40 degrees C followed with two water rinses. PMID- 10379032 TI - Particle size separation via soil washing to obtain volume reduction. AB - A pilot-plant study was performed using a soil washing pilot plant originally designed by the Environmental Protection Agency (EPA) to demonstrate scale-up and potential full-scale remediation. This pilot plant named VORCE (Volume Reduction/Chemical Extraction) was modified to meet the specific requirements for treatment of the Formerly Utilized Sites Remedial Action Program (FUSRAP) and a Department of Energy site soils. After a series of tests on clean soils to develop operating parameters and system performance, the machine was used to treat soils, one contaminated with Thorium-232 and the other with Cesium-137. All indicate that soil washing is very promising for volume reduction treatment. In addition, cost data was generated and is given herein. PMID- 10379033 TI - Selective removal of plutonium 238 from a canal sediment using a carbonate chelant soil washing technology (ACT*DE*CON). AB - The Mound laboratory site in Miamisburg, OH, a former plutonium processing facility, contains approximately 40000 yd(3) (30,580 m3) of plutonium- and thorium-contaminated soils and sediments at levels that require remediation. Existing applicable remediation technologies are unsatisfactory, because they are expensive and do not provide volume reduction. ACT*DE*CON is a chemical soil leaching technology for the treatment of soils that utilizes contaminant dissolution via dilute selective solutions to remove radionuclides. In bench scale tests, process parameters were developed for the optimal treatment of the Miami Erie Canal soil at the Mound site, combining the maximum plutonium removal with an acceptable amount of soil dissolution and minimizing the costs of reagents. Parameters evaluated included soil to extractant mass ratio, temperature, rinse solution composition, kinetics, and the application of several dewatering aids. Plutonium removal rates of >95% were achieved, and the residual plutonium in the treated soil proved to be very immobile-confirming that the process had removed the most accessible species of the radionuclide. Currently being tested at Mound is an engineering scale-up that includes an attrition scrubber, a counter-current extractor, and a reverse osmosis system. Economic evaluations based on bench-scale results put the treatment cost at US$278/yd(3) (US$364/m3), compared to US$350/yd(3) (US$458/m3) for the 'box-and-bury' baseline alternative treatment system. PMID- 10379034 TI - Full-scale and pilot-scale soil washing. AB - The purpose of this paper is to describe soil washing and to present results obtained from pilot-scale and full-scale projects. The soil washing system to be described is a water-based physical separation process which relies on traditional physical and chemical extraction and separation processes for removing a broad range of organic, inorganic, and radioactive contaminants from soil. Although soil washing is becoming more accepted as a treatment technology in the United States, limited experience in field application still appears to be a barrier to more widespread implementation. This paper will attempt to overcome some of those barriers by describing the system and its applications, and providing case histories of successful experiences in full-scale and pilot-scale field operations. Both levels of operations have been very successful, and confirm the viability of soil washing for treating contaminated soils. PMID- 10379035 TI - Laboratory treatability testing of soils contaminated with lead and PCBs using particle-size separation and soil washing. AB - A soil treatability study was conducted using particle-size separation and soil washing to reduce the volume of material contaminated with polychlorinated biphenyls (PCBs) and lead at a Superfund site. Soil washing using surfactant was effective at removing 95% of PCBs into fine material and residual wash water. Results indicate that almost 80% of the material contaminated with up to 140 mg/kg PCBs could be treated to concentrations below 10 mg/kg using soil washing with surfactant. There did not appear to be a difference in lead removal using either particle size separation or soil washing, although the lead data have high uncertainty because of soil heterogeneity. Lead concentrations in soil were reduced from as high as 1700 to < or =150 mg/kg and from 560 to < or =220 mg/kg in about half of the material using particle size separation. PMID- 10379036 TI - Chelant extraction of heavy metals from contaminated soils. AB - The current state of the art regarding the use of chelating agents to extract heavy metal contaminants has been addressed. Results are presented for treatability studies conducted as worst-case and representative soils from Aberdeen Proving Ground's J-Field for extraction of copper (Cu), lead (Pb), and zinc (Zn). The particle size distribution characteristics of the soils determined from hydrometer tests are approximately 60% sand, 30% silt, and 10% clay. Sequential extractions were performed on the 'as-received' soils (worst case and representative) to determine the speciation of the metal forms. The technique speciates the heavy metal distribution into an easily extractable (exchangeable) form, carbonates, reducible oxides, organically-bound, and residual forms. The results indicated that most of the metals are in forms that are amenable to soil washing (i.e. exchangeable+carbonate+reducible oxides). The metals Cu, Pb, Zn, and Cr have greater than 70% of their distribution in forms amenable to soil washing techniques, while Cd, Mn, and Fe are somewhat less amenable to soil washing using chelant extraction. However, the concentrations of Cd and Mn are low in the contaminated soil. From the batch chelant extraction studies, ethylenediaminetetraacetic acid (EDTA), citric acid, and nitrilotriacetic acid (NTA) were all effective in removing copper, lead, and zinc from the J-Field soils. Due to NTA being a Class II carcinogen, it is not recommended for use in remediating contaminated soils. EDTA and citric acid appear to offer the greatest potential as chelating agents to use in soil washing the Aberdeen Proving Ground soils. The other chelating agents studied (gluconate, oxalate, Citranox, ammonium acetate, and phosphoric acid, along with pH-adjusted water) were generally ineffective in mobilizing the heavy metals from the soils. The chelant solution removes the heavy metals (Cd, Cu, Pb, Zn, Fe, Cr, As, and Hg) simultaneously. Using a multiple-stage batch extraction, the soil was successfully treated passing both the Toxicity Characteristics Leaching Procedure (TCLP) and EPA Total Extractable Metal Limit. The final residual Pb concentration was about 300 mg/kg, with a corresponding TCLP of 1.5 mg/l. Removal of the exchangeable and carbonate fractions for Cu and Zn was achieved during the first extraction stage, whereas it required two extraction stages for the same fractions for Pb. Removal of Pb, Cu, and Zn present as exchangeable, carbonates, and reducible oxides occurred between the fourth- and fifth-stage extractions. The overall removal of copper, lead, and zinc from the multiple-stage washing were 98.9%, 98.9%, and 97.2%, respectively. The concentration and operating conditions for the soil washing extractions were not necessarily optimized. If the conditions had been optimized and using a more representative Pb concentration (approximately 12000 mg/kg), it is likely that the TCLP and residual heavy metal soil concentrations could be achieved within two to three extractions. The results indicate that the J-Field contaminated soils can be successfully treated using a soil washing technique. PMID- 10379037 TI - Evolution of the biopharmaceutics classification system (BCS) to oral modified release (MR) formulations; what do we need to consider? AB - The Biopharmaceutics Classification System (BCS) has provided a mechanistic framework for understanding the concept of drug absorption in terms of permeability and solubility. Regulatory guidance for Immediate Release (IR) products incorporating the BCS has appeared over recent months. Extrapolation of the approach to oral Modified Release (MR) formulations is therefore inevitable, however, further evolution of our thinking is first required. MR products can deliver drug throughout the gastrointestinal tract and it is important that we recognise there is significant heterogeneity in permeability, lumenal pH/contents and gut wall metabolism from stomach to colon. PMID- 10379038 TI - Controlling drug delivery across the placenta. AB - A challenge in modern drug therapy is to develop strategies for safer and more selective targeting of drug delivery in pregnancy. Specifically, approaches are needed that would restrict unnecessary drug exposure to either mother or fetus. There is evidence emerging that indicates the placenta does express natural transport and metabolism processes that function to control drug and nutrient distribution between the mother and fetus. Further, in vitro techniques developed in the past 10 years now provide some of the tools necessary to elucidate transport and metabolism processes typical of the human placenta. As a consequence, pharmaceutical scientists are in a position to contribute significantly to the design and development of drugs for pregnancy. PMID- 10379039 TI - Mucus as a barrier to the permeability of hydrophilic and lipophilic compounds in the absence and presence of sodium taurocholate micellar systems using cell culture models. AB - A mucus secreting CaCo-2-Ht29GlucH cell co-culture model was characterised and used to examine the influence of mucus as a barrier to the transport of hydrophilic and lipophilic compounds in the absence and presence of sodium taurocholate micellar (NaTC) systems. TEER measurements and permeability studies using the hydrophilic markers (mannitol, polyethylene glycols (PEGS) 900 and 4000) indicated that the paracellular permeability of the co-culture model was greater than that of the CaCo-2 model. At pH 7.4, no difference in the transport of a model lipophilic drug, dextropropoxyphene, was observed between the two models. However, at pH 4.5, when the drug was highly ionised the transport was significantly lower across the co-culture monolayers. NaTC micellar systems appeared to affect the different cell culture models in the order CaCo-2>CaCo-2 Ht29GlucH>Ht29GlucH. Following removal of the mucus layer by incubation with the mucolytic agent, N-acetyl-l-cysteine, the absorption enhancing potential of NaTC micellar systems was increased in the co-culture model. PMID- 10379041 TI - Gynostemma pentaphyllum: identification of major sapogenins and differentiation from Panax species. AB - Four main dammarane-type aglycones of gypenosides, extracted from the aerial parts of Gynostemma pentaphyllum were identified by gas chromatography-mass spectrometry. By detecting these aglycones as well as the aglycones of ginsenosides, a difference in sapogenin composition between Gynostemma pentaphyllum and Panax species was observed, which can be used in the differentiation of these plant drugs. PMID- 10379040 TI - Sorbitan monostearate/polysorbate 20 organogels containing niosomes: a delivery vehicle for antigens? AB - Multi-component organogels formed using the non-ionic surfactant sorbitan monostearate as gelator have been formulated to contain niosomes. The purpose of this study was to evaluate the potential of these vesicle-in-water-in-oil (v/w/o) gels as delivery vehicles for vaccines. Bovine serum albumin (BSA) and haemagglutinin (HA) were used as model antigens in depot and immunogenicity studies respectively. The complex gels were prepared by the addition of a hot (60 degrees C) aqueous niosome suspension (v/w) to the sol phase (o, an organic solution of the gelator); a vesicle-in-water-in-oil (v/w/o) emulsion was produced which cools to an opaque, semi-solid, thermoreversible v/w/o gel. Light microscopy of the organogel revealed that the microstructure consists of a tubular network of surfactant aggregates in the organic medium, the niosome suspension being dispersed in these surfactant tubules. Therefore, in such gels, the vaccine is thought to be entrapped in the niosomes, themselves located within the sorbitan monostearate tubular network in the organic medium. In vivo, a depot effect was observed following intramuscular administration of the gel containing the entrapped bovine serum albumin, cleared from the injection site over a period of days. The relatively short-lived nature of the depot was thought to arise due to interactions between the gel and the local interstitial fluid which results in gel disintegration in situ. Thus, the niosomes containing antigens are believed to be released from the organic gel. Immunogenicity studies showed that the v/w/o gel as well as one of the controls, the water-in-oil (w/o) gel, possess immunoadjuvant properties and enhance the primary and secondary antibody titres (of total IgG, IgG1, IgG2a and IgG2b) to haemagglutinin antigen. As far as humoral immunity is concerned, the w/o gel showed stronger immunoadjuvant properties compared to the v/w/o gel, being effective at a lower antigen dose i.e 0.1 microg HA. PMID- 10379042 TI - Influence of neutron activation factors on the physico-chemical properties of suppositories and their excipients. AB - The effect of the neutron activation factors, i.e., admixture of samarium oxide (Sm2O3) and irradiation time, on the physico-chemical properties of the raw materials and the in vitro dissolution and disintegration of hydrophilic and lipophilic suppositories was investigated. It was possible to expose the pure bases and the model drugs (5-aminosalicylic acid [5-ASA] and ropivacaine hydrochloride) to 1 min of neutron irradiation in a flux of 1.1.1013 n cm-2s-1. The dissolution and disintegration of the corresponding suppositories showed that the physico-chemical properties and the fraction of incorporated drug together with the lipophilic/hydrophilic nature of the base were important factors. Sm2O3 increased the disintegration time of hydrophilic suppositories containing 5-ASA, while the dissolution of both drugs from these formulations remained unchanged. Sm2O3 did not alter the disintegration time of the lipophilic formulations, but it reduced the dissolution of both drugs from these suppositories. Irradiation induced different behaviour in the different bases. PMID- 10379043 TI - Distribution of ciprofloxacin in the isolated rat lung in the presence and absence of tissue oedema. AB - A series of experiments with the isolated perfused rat lung was carried out to study the distribution of ciprofloxacin in this tissue under different experimental conditions; the influence of drug administration rate and the presence of oedema were evaluated by comparison of the statistical moment and distribution coefficient values as well as the unit disposition function (UDF) profiles in the tissue. The polyexponential character of the outflow perfusate curves indicates a distribution behaviour which does not correspond to the 'well stirred model' but rather to the existence of some type of diffusion barriers and/or tissue binding. The presence of oedema in the tissue, induced by insufficient oxygenation of perfusate, significantly increases the distribution coefficient of ciprofloxacin and leads to relevant modifications in the unit disposition function (UDF) of the isolated lung, while administration rate does not significantly affect the kinetic behaviour of the drug in this tissue. PMID- 10379044 TI - Longitudinal versus radial effects of hydroxypropylmethylcellulose on gastrointestinal glucose absorption in dogs. AB - Many water soluble fibers have been shown to favorably affect the postprandial glucose profile in humans. Hydroxypropylmethylcellulose (HPMC), a fiber which has been shown to increase glucose tolerance in dogs and noninsulin dependent diabetics, was chosen to study the luminal interactions which mediate this effect. The ability of HPMC to influence upper gastrointestinal (GI) viscosity, transit, and water flux of 5% and 20% glucose solutions was studied in five female dogs fistulated at the proximal duodenum and/or midjejunum. HPMC elevated intraluminal viscosity, with a linear relationship existing between input and luminal viscosity. The ability to modify intraluminal viscosity was greater for isoosmotic (5%) glucose solutions than for hyperosmotic (20%) glucose solutions. HPMC also modified the transit profile of isoosmotic (5%) glucose solutions at midgut by both increasing the lag times before the onset of chyme recovery from 5.5+/-3.1 min to 9-55 min (depending on the viscosity of the administered solution) and decreasing the first-order transit rate constants from 0.115+/-0.07 min-1 to 0.014-0.035 min-1. By contrast, the transit profile of hyperosmotic (20%) glucose solutions was not significantly affected. Net cumulative water flux across the gut wall was not significantly affected in either case by the presence of HPMC. These results, in combination with the amount of glucose recovered from midgut fistula, suggest that following the administration of glucose solutions, HPMC effects on blood glucose levels are mediated by mechanisms which relate to the increased intraluminal viscosity but vary according to the input glucose load. For isoosmotic glucose loads, both the decreased upper GI transit rate and hindered radial movement play a role. Although HPMC modifies glucose absorption from hyperosmotic solutions, this study shows that luminal effects occurring before midgut are modest. PMID- 10379045 TI - Effect of calcium concentration, hardening agent and drying condition on release characteristics of oral proteins from calcium pectinate gel beads. AB - Pectin has been investigated for its ability to produce solid calcium pectinate gel (CPG) beads containing bovine serum albumin (BSA). Several factors can influence the properties and release characteristics of the CPG beads. In this study, the effect of calcium concentration, hardening agent and drying condition on the encapsulation and release characteristics of BSA from the matrix gel beads made of calcium pectinate were studied. BSA release studies under conditions mimicking mouth to colon transit have shown that calcium pectinate protects the drug from being released completely in the physiological environment of the upper gastrointestinal tract, and is susceptible to the enzymatic action with consequent drug release. In addition, the release of BSA from CPG beads was strongly affected by calcium concentration and drying condition. However, the release was not particularly affected by the presence of hardening agent at the concentration of 1% or lower. Since the release of BSA as a model protein drug could be controlled by the regulation of the preparation conditions of CPG beads, the CPG beads may be used for a potential oral controlled release system for protein drugs. PMID- 10379046 TI - The neuroendocrine immune basis of rheumatic diseases. PMID- 10379047 TI - Distribution of activated T cells migrating through the body: a matter of life and death. AB - The preferential distribution of lymphocyte subsets in tissues is attributed to a selective lymphocyte-endothelium interaction during entry. However, proliferation and death within the tissue, and exit from the tissue, might also play a role. Here, Jurgen Westermann and Ulrike Bode provide evidence that preferential survival in the tissue of initial stimulation is the major factor in the preferential distribution of activated T cells. PMID- 10379048 TI - Cytokine/chemokine cascades in immunity to tuberculosis. AB - The relationship between acquired specific resistance and delayed-type hypersensitivity (DTH) in immunity to tuberculosis has long been a topic of debate. Here, Ian Orme and Andrea Cooper propose that the events are separate mechanisms; protection is cytokine driven and initially controls the infection, whereas DTH is primarily chemokine driven and functions to wall off the infection and prevent further dissemination. PMID- 10379049 TI - CD5 expression in human B-cell populations. AB - The origin of CD5+ B cells remains controversial. The differential response to ligation of CD5 resulting in apoptosis or proliferation provides insight into its roles in distinct human B cells. Here, Pierre Youinou, Christophe Jamin and Peter Lydyard review current knowledge of B-1 and B-2 cells, and propose that CD5 has different functions when expressed by different B-cell subpopulations. PMID- 10379050 TI - Atopic disorders: a default pathway in the absence of infection? AB - Atopic disorders, such as asthma, are increasingly prevalent in the developed world. Recent epidemiological and experimental data suggest that some infectious diseases prevalent in the developing nations can inhibit the development of allergic disorders. Here, Klaus Erb reviews the data and presents a model showing how a steady decline of infectious diseases could account for an increase in atopic disorders. PMID- 10379051 TI - Efficacy of adhesive interactions in pig-to-human xenotransplantation. AB - Successful xenotransplantation depends on many factors, one being the interactions of cross-species adhesion molecule-ligand pairs. Depending on the approach used to facilitate xenotransplantation, these interactions can play differing roles. Here, Andre Simon, Anthony Warrens and Megan Sykes review the existing information on pig-to-human adhesive interactions and its implication for different approaches to pig-to-human xenotransplantation. PMID- 10379052 TI - T-cell function in newborn mice and humans. AB - Neonates mount poor immune responses, and it has been assumed that neonatal T cells differ qualitatively from adult T cells. Here, Becky Adkins discusses this issue in the light of recent data indicating that T cells in neonates are developmentally mature in their capacity to mount protective Th1-type and cytotoxic T lymphocyte responses. PMID- 10379053 TI - Granuloma formation and tuberculosis transmission in HIV-infected patients. PMID- 10379054 TI - Granulomatous inflammation and transmission of infectious disease. PMID- 10379055 TI - Granulomatous inflammation and transmission of infection - reply PMID- 10379056 TI - [Progressive multifocal leukoencephalopathy: morphologic possibilities of diagnostic classification methods and in situ hybridization]. AB - Progressive multifocal leucoencephalopathy is caused by infection with JC virus. The disease affects patients with immunodeficiencies, hematologic diseases, and patients treated with radiotherapy. The disease is characterised by foci of demyelinisation with atypical astrocytes and oligodendrocytes. Oligodendrocytes contain typical intranuclear inclusions. Progressive multifocal leucoencephalopathy and its verification is presented in three cases. Two patients died of progression of a malignant neoplasm and the leucoencephalopathy was a complication of the malignancy. The third case was a biopsy specimen taken from the brain of a patient who received a renal transplant. The material of all patients was analysed by light and electron microscopy, and in situ hybridisation with a probe specific for JC virus. In situ hybridisation proved to be the most specific and a simple method to demonstrate the infection in all cases. It is useful in instances in which the histologically detectable lesion is not characteristic, and in cells in which the conventional histologic methods fail to reveal the intranuclear inclusions of JC virus. PMID- 10379057 TI - [Immunohistochemical analysis of renal angiomyolipoma]. AB - Immunohistochemical analysis of 6 cases of renal angiomyolipoma not related to tuberous sclerosis proved the same rate of expression of HMB 45 and NKI/C3 as well in epithelioid smooth muscle cells of the tumours. Beyond this, an unspecific expression of alpha 1-antitrypsin and alpha 1-antichymotrypsin was found which has not been mentioned in the literature up to now. PMID- 10379058 TI - [Sarcomatous chromophobe cell renal carcinoma. 2 case reports]. AB - Chromophobe cell renal carcinoma is a relatively rare primary tumour of the kidney. We present two cases of sarcomatoid transformation in this type of renal cell carcinoma. Described transformation in chromophobe renal cell carcinoma has been reported previously only in four publications. We present a very unusual aggressive recurrence of one of our cases and introduce the first case of sarcomatoid carcinoma arising from chromophobe cell renal carcinoma of the kidney with rhabdomyoblastic differentiation. PMID- 10379059 TI - [A testicular tumor with features of a sex cord tumor with annular tubules and calcified large-cell Sertoli tumor]. AB - An unusual testicular tumour with features of both sex cord tumour with annular tubules and large-cell calcifying Sertoli cell tumour is described. The tumour occurred in a 20-year-old man in the left testis as a 1 cm-measuring well circumscribed solitary lesion. Histologically, it showed sertoliform annular tubules with included eosinophilic nodules as well as isolated large cells and cords to sheets of the cells lying in a sclerosing stroma. A few psammomatous microcalcifications were seen. In contrast to similar cases reported before, the patient did not have any signs of Peutz-Jeghers syndrome, feminization or other complex dysplastic syndromes. The case indicates that tumours of this type may occur in phenotypically normal patients like pure large cell calcifying Sertoli cell tumours or ovarian sex-cord tumours with annular tubules. It further supports a close histogenetic relationship between these two entities. PMID- 10379060 TI - [Systemic embolization of mucous material]. AB - Systemic embolisation of mucous substance occurs as a rare complication of mucinous carcinomas. A case of primary carcinoma of stomach is described with embolisation of mucus into cerebral, renal, pulmonary and myocardial vessels. Vascular blockade by mucus caused multiple partly haemorrhagic microinfarctions of the brain tissue. Beyond this, investigated tumour samples did not show any signs of angioinvasion and the production of mucus was far from massive. PMID- 10379061 TI - [Heterotopic chondro-osteoplastic metaplasia of the subclavian vein wall after long-term catheterization. Case report]. AB - A 59-year-old man admitted for i.m. had a 3-way cannula inserted into the right subclavian vein for 31 days. Autopsy revealed a circular firm thickening of the venous wall. Histology identified it as chondroid and osteoid tissue replacing adventitial connective tissue. Unusual chondroosteoplastic metaplasia not described up to now is discussed as about its causes. PMID- 10379062 TI - [Immunohistochemical detection of cytokine receptors on cryostat tissue sections]. AB - We have studied tissue expression of the cytokine receptors using a high sensitivity biotin-streptavidin system on cryostat sections. We used a panel of monoclonal antibodies from the 6th International Workshop on Human Leukocyte Differentiation Antigens, namely CD25 (IL-2R alpha), CD95 (FAS antigen), CD116 (GM CSFR), CD117 (SCFR), CD120 alpha (TNFR I), CD120b (TNFR II), CD121a (IL-1R I), CDw123 (IL-3R), CD124 (IL-4R), CD126 (IL-6R), CD127 (IL-7R), CDw128 (IL-8R), CD130 (gpl130), CD131 (IL-3R), CD132 (IL-2R gamma), CD134 (OC-40), CD135 (FLT3/FLK2). Examined tissues (lymph nodes and spleens) were obtained from 12 patients with folicular non-Hodgkin's lymphoma, periferal T non-Hodgkin's lymphoma, B lymphoma, myeloma, Hodgkin's disease, two cases of T cell rich B lymphoma, autoimmune haemolytic anemia and two cases of rudimentary trombocytopenic purpura. Our results indicate that immunohistological technology using native tissues on cryostat sections, monoclonal antibodies and the visualisation with biotin-streptavidin is a particularly suitable supplementary staining procedure for detection of the cytokine receptors in tissues. PMID- 10379063 TI - [Fatal injuries in bicyclists]. AB - An analysis of 58 cases of fatal bicycle accidents during the years 1991-1996. Substantial prevalence of males was a common feature among deceased persons especially in the 21-40-year and 61-70-year age groups. More fatal bicycle accidents occurred in evening hours between 6 and 12. The proportion of alcohol influence among dead cyclists was standardly high and the usage of protective helmet was not found. Further analysis concerned the types of injuries after fall on a firm base, after collision with a solid subject or with means of transformation. Craniocerebral injury was the cause of death in 65.5% of cases. Because of increase in popularity of bicycle as simple means of transportation a further increase of fatal cyclist accidents can be presumed requiring precise follow-up by forensic medicine. PMID- 10379064 TI - [Determination of the ABH blood group system in the placenta]. AB - Blood group substances A, B, H were detect in placental tissue paraffin cuts (after 10% formaline fixation) by immunohistochemistry using indirect immunoperoxidase-two-layers technology or biotin-streptavidin complex signed by alkaline phosphates. Both fetal and maternal part of placenta are to be investigated which enables their group markers to be identified. Immunohistochemical detection of the A, B, H blood group substances in placenta can give conclusion about the blood group of fetus and of fetus and of mother as well. PMID- 10379065 TI - [Evaluation of the Sarstedt S-Monovette collection kit in the toxicologic evaluation of ethanol and other volatile substances in the blood]. PMID- 10379066 TI - [Current data on bipolar disorders, epilepsy and spasm. 4th Neuro-Forum, Dresden]. PMID- 10379067 TI - [Dielectric spectroscopy for noninvasive examination of corneal tissue]. AB - Dielectric spectroscopy is a non-invasive contact technique that permits the in vivo measurement of the specific electrical properties of biological tissue induced by an external electrical field. Permittivity, relaxation time and specific conductivity as a function of corneal hydration (wet weight/dry weight) and temperature were measured in 10 porcine corneas. Variation of tissue hydration has a minor influence on the signal, with a significant variation of the signal being detectable only for relatively dry tissue. A much greater influence was found for temperature, in particular on relaxation times. Dielectric spectroscopy provides us with an opportunity to detect structural, in particular temperature-induced, changes in living tissue. In the frequency range investigated, hydration has only a small influence on the dielectric properties of the tissue. PMID- 10379068 TI - [Critical evaluation of indications for the holmium:YAG laser and the neodymium:YAG laser in orthopedic surgery based on an in vitro study]. AB - This is an in vitro study of the biophysical effects of holmium:YAG and neodymium YAG lasers that was prompted by the poor clinical results obtained with lumbar percutaneous laser discus decompression (PLDD). In the absence of adequate cooling, ablation of tissue with the holmium:YAG laser causes thermal damage to the surrounding tissues. Utilizing the immediate colour-independent laser coupling effect, the holmium:YAG laser removes soft and hard tissue immediately. The low tissue penetrating power (max. 0.32 mm), together with the use of irrigation, avoids thermal problems, and this laser type with its high pulse energy and frequency is to be recommended for arthroscopic surgery. In contrast, the effects of the neodymium:YAG laser are highly dependent on tissue colour. Using this laser on light-coloured tissue only diffuse warming but no ablation of soft tissue was often seen. The depth of tissue penetration seen in our study was 0.58 mm, but is greatly dependent on the duration of application, and is much larger with long application times. In conclusion, we believe that the neodymium:YAG laser is more suitable for percutaneous intradiscal procedures than the holmium:YAG laser. For arthroscopic surgery, the holmium:YAG laser will be the better choice. The effect of each type of laser depends not only on its physical properties, but also on tissue properties (light or dark-coloured, thermal conductivity) and duration of application. PMID- 10379069 TI - [Detection of cerebral embolism using transcranial Doppler sonography: can artifacts be reliably recognized?]. AB - A requirement for the use of TCD for the detection of emboli in the field of cardiac and vascular surgery is the reliable differentiation between true emboli and artifacts. In ten healthy volunteers we carried out a study to establish the method with which artefacts can most reliably be identified. Automatic detection of increasing signal intensity misinterpreted 14% of all artifacts as emboli; 1.7% of all artifacts sounded suspicious for embolism, and 0.6% met the classical criteria of an embolus. Using simultaneous recording of the flow signal in two sections of the middle cerebral artery, all artifacts were identified on the basis of their simultaneous manifestation. Reliable intra-operative differentiation of emboli from artifacts requires attentive, continuous acoustic and visual analysis of signals by an experienced investigator familiar with the surgical procedure. The introduction of a multiple-depth algorithm might significantly improve the automatic detection program. PMID- 10379070 TI - [New instruments for preparation of the prosthesis socket and primary stability of the acetabular Press-fit cups]. AB - The aim of the present study was to develop surgical instruments necessary to achieve a precisely reamed surface and stable initial fixation. The instruments used to prepare the socket were a gauge-drill guide, a liner for the spigot hole and two spigotted reamers of different design and indentation intended to achieve a precisely reamed surface while preserving subchondral bone. For each reamer we implanted in a synthetic hip model 10 uncemented cups with 2 mm press-fit and loaded at 2.4 kN in the physiological axis (Mod. 8501, Instron, Canton, MA, USA). The micromotion between implant and bone socket was measured using an inductive micrometric measuring system (MultiNCDT-500, Micro-Epsilon, Ortenburg, Germany) and compared with that seen after using conventional instruments. The use of the new reamer of elliptical design significantly reduced the standard deviations of the measured values (p < 0.01 at the ischium and pubis) and also reduced maximum movement (p < 0.01 at the ischium); at the same time, all the components showed overall limited movement (< 150 microns at the ischium, pubis and ilium) under maximum loading (2.4 kN). Manufacturing tolerances, the quality and wear of the instruments, acetabular bone stock and surgical technique all impact on the degree of press-fit obtainable at surgery. The results of our study show that press-fit and initial stability can be optimized by using adequate instruments to prepare the socket. PMID- 10379071 TI - [Forces acting on foot soles during stair climbing in healthy probands and in patients with coxarthrosis]. AB - We investigated the contact forces acting on the sole of the foot of healthy persons and coxarthrosis patients climbing and descending stairs. The sole contact forces were determined using an experimental set-up comprising a stair construction provided with an integrated measuring step. RESULTS: In healthy subjects, the forces acting on the soles of the feet while climbing stairs were found to be 1.2 times their body weight. With regard to descending stairs, a distinction must be made between "hard" and "soft" walkers. In the case of "hard" walkers, the forces acting on the soles may be as much as 2.6 times body weight. These forces can be reduced by the wearing of shock-absorbing shoes. CONCLUSION: In coxarthrosis and prostheses-bearing patients, all movements are executed more slowly when climbing or descending stairs, so that only small dynamic forces arise. The greatest loads are about 1.2 times the patient's own weight. In these patients, an effective reduction by shockabsorbing footwear is not possible. PMID- 10379072 TI - [3-dimensional registration of micro-movements of vertebral implants by a specially adapted measuring system using magnetic transducers]. AB - The aim of this study was to develop a biomechanical experimental set-up to quantify motion of ventrally inserted spinal implants at the implant/bone interface. The model we used was the vertebral column of the calf. Lumbar vertebrae L2 to L4 were "instrumented" with a screw-rod system. The adjacent vertebrae L1 and L5 were connected to a servohydraulic testing machine and axial compression applied. Shortening of the specimen and three-dimensional movement of the most cranial implant relative to the bone was recorded using 3 electromagnetic transducers. 100,000 cycles of axial loading varying between -0.5 kN and -1 kN were applied. Static shortening of the specimen of 8.5 mm and an elastic movement of 180 to 280 microns were measured. The greatest amplitude of single-plane motion was recorded in the sagittal plane in both static and elastic modes. Implant motion within each cycle was recorded accurately as load displacement curves within a range of 1.35 to 30 microns. With this test set-up, primary stability of different spinal implant systems can be compared. The use of electromagnetic transducers permits three-dimensional implant motion analysis even when only a mono-axial servohydraulic testing machine is available. PMID- 10379073 TI - Concepts and terminology in ethnicity, race and health: be aware of the ongoing debate. AB - Although the expansion of the scientific literature in the field of ethnicity, race and health is welcome, it has been weakened by the diverse and inconsistent terms used to describe a group or individual's ethnicity. Further, a clear definition of what is meant by the terms ethnicity and race in publications is often lacking, making it difficult to compare studies. This problem is leading to much debate in the USA and the UK. Journal editors and researchers need to be aware of these debates, and actively involved in resolving problems and raising standards. PMID- 10379074 TI - Child care and oral health. PMID- 10379075 TI - The dental care of children. PMID- 10379076 TI - Radiograph selection. PMID- 10379077 TI - Sedation drug use. PMID- 10379078 TI - Sedation is not the only answer. PMID- 10379079 TI - Minority ethnic groups in health care professions. PMID- 10379080 TI - Eagle's syndrome: an unusual cause of a clicking jaw. AB - Calcification of the stylohyoid ligament is a well recognised radiographic finding in dental practice. Fortunately, affected individuals seldom develop symptoms. We report a case of a patient whose main complaint was a loud click following jaw movement. This unusual presentation has not been described before and should be considered in the differential diagnosis of 'clicking jaw'. PMID- 10379081 TI - Tooth wear in the child and the youth. AB - Tooth surface loss is an increasing problem in younger individuals. Preventive strategies are essential while adhesive dentistry should be used whenever possible if restoration is necessary. PMID- 10379082 TI - Osseointegrated implants in post-surgical prosthodontic rehabilitation. AB - The advent of osseointegrated implants and their use in dentistry has been well documented and continues to become more and more a part of conventional dental treatment. Apart from their use in simple situations they can prove invaluable when there is little hope of success with the other options. Two cases are presented here that involve multiple tooth loss as a result of surgical intervention and treatment of pathological lesions. PMID- 10379083 TI - A psychodynamic understanding of the dentist-patient interaction. AB - This paper describes various elements of the dentist-patient relationship, how they interact and illustrates them with case studies. PMID- 10379084 TI - Factors influencing nerve damage during lower third molar surgery. AB - OBJECTIVE: To investigate relationships between pathology, eruption status, age, anaesthetic modality and nerve damage during lower third molar surgery. DESIGN: Single centre prospective study. SETTING: Oral surgery out-patient clinics. SUBJECTS: 367 patients unselected for age, gender or social class, scheduled for lower third molar removal. At 1 week, any evidence of iatrogenic nerve damage was recorded. Patients with altered lingual and/or labial sensation were followed up for 6 months. RESULTS: 718 lower third molars were removed from 250 males and 117 females. 96 removals (13.4%) were associated with altered lingual, labial or buccal sensation. There were no significant associations between nerve damage and eruption status, age and pre-operative pathology. There was a highly significant difference in the incidence of nerve damage between LA removal (3%) and GA removal (18%) (chi-squared = 17.18; f = 2; P < 0.01) but no significant associations between surgical difficulty and nerve damage within each of the two groups. CONCLUSIONS: Lingual and inferior alveolar nerve damage was five times more frequent when lower third molars were removed under general anaesthesia rather than local anaesthesia. This could not be explained in terms of surgical difficulty, pre-operative pathology, age or anatomical position. PMID- 10379085 TI - Dental service use and the implications for oral cancer screening in a sample of Bangladeshi adult medical care users living in Tower Hamlets, UK. AB - AIM: To assess the use of dental services, barriers to uptake of dental care and attitudes to regular dental examinations and the prevalence of tobacco and paan chewing habits in a group of Bangladeshi medical care users. DESIGN: Multi-centre cross-sectional study. SETTING: Four general medical practices' waiting areas in Tower Hamlets. SUBJECTS: Bangladeshi adults aged 40 years and over. INTERVENTION: An interview schedule. MAIN OUTCOME MEASURES: The prevalence of tobacco smoking and paan chewing with or without the addition of tobacco. The use of dental services, barriers to the use of dental services and attitudes to regular dental examinations. RESULTS: Results were obtained from 158 subjects (response rate 85%). 25% of the whole sample had never visited a dentist. These were significantly (P < 0.05) more likely to be women, who also thought regular check ups were of little value. In their use of health services 73% experienced language difficulties. 33% of the sample were tobacco smokers. Paan was chewed by 78% of the sample with significantly (P < 0.05) more females than males adding tobacco to their quid and chewing more frequently than males. CONCLUSION: There are considerable barriers to be overcome if dental practices are to be the site for oral cancer screening and oral health promotion in this population. There are sex differences in reported behaviour and attitudes about use of dental services and in tobacco and paan use in this Bangladeshi sample. Further research is needed to establish why this ethnic minority attend general medical practices but not general dental practices. PMID- 10379086 TI - Regional odontodysplasia: an unusual case with a conservative approach. AB - A case of a 14-year-old male with regional odontodysplasia is reported. In this presentation many atypical clinical and radiographical features of this condition are present. The chief complaint of the patient was the enlargement of the gingiva and, according to the literature, inflammatory processes are the main reason why patients look for care. Moreover, there was no radiographic evidence of unerupted teeth in this report. The functional and psychological benefits of the conservative approach are emphasised. PMID- 10379087 TI - Dental pathology in ancient Mesopotamia. PMID- 10379088 TI - Ut Dicunt Medici: medical knowledge and theological debates in the second half of the thirteenth century. PMID- 10379089 TI - Public health versus private practice: the contested development of compulsory infectious disease notification in late-nineteenth-century Britain. PMID- 10379090 TI - Revisiting and rethinking the rewriting of nursing history. PMID- 10379091 TI - An early report of familial bronchiectasis. PMID- 10379092 TI - Acute hepatitis B in injecting drug users is increasing as initiative to improve vaccine coverage begins. PMID- 10379093 TI - Chance finding of hepatitis B e antigen carriage in pregnant woman highlights need for antenatal screening, and vaccination of health care workers. PMID- 10379094 TI - Depolarising the 'broadened' and 'back-to-basics' relief models. AB - Prompted by the calls in recent years to link relief practice to peace-building, development, or both, several conferences and papers have recently mounted a strong critique, seeing these 'broadenings' of relief as 'eroding' or 'corrupting' core humanitarian principles and playing into neo-isolationist agendas to slash humanitarianism. This paper argues that whereas these critiques have important goals in mind when they encourage a concentration on the basics of relief, there has been a loss of subtlety in the ensuing debate. Considering each element of the debate in turn, the paper argues that there is more common ground between 'new' and 'old', 'broadened' and 'basics' relief than at first appears. In concluding, it is argued that further research on key questions, and an openness to hear all perspectives will get us further than entrenched positions and rallying cries. PMID- 10379095 TI - An ombudsman for humanitarian assistance? AB - The early phases of a project to design and make operational an ombudsman for humanitarian assistance (HAO) are described in this paper. Beginning with a brief historical overview of the ombudsman concept, it then outlines seven key features of a potential HAO that were identified in the initial feasibility study. The main conclusion from the feasibility study was that, in principle, it is possible to design an HAO by adapting the operational frameworks of existing ombudsman schemes so as to match the needs of the humanitarian sector. Although this seems possible in theory, there still remain some major challenges requiring practical testing in a pilot phase. The most fundamental of these is how to enable the beneficiaries of aid to make their voices heard and register their views on the management of the emergency that is affecting them. Important also, are the issues of ensuring an international jurisdiction and finding sustainable ways of financing the scheme. Finally, the paper alludes to the framework of the pilot itself and how to address the main challenges ahead. PMID- 10379096 TI - Humanitarianism, pluralism and ombudsmen: do the pieces fit? AB - A variety of codes and standards for humanitarian assistance have been put forth in recent years. Many NGOs have agreed to abide by these codes. There is uncertainty, however, about if and how these codes are actually being put into practice. Have we moved from words to action? One response to this concern has been a proposal to establish a humanitarian ombudsman. This paper analyses two choices facing an eventual ombudsman: whether to attempt to take punitive actions to enforce the codes and standards, or whether instead to facilitate agencies' own internal efforts to improve accountability to their beneficiaries. It proposes a pluralistic approach, wherein a variety of methods, structures and local perceptions are accepted as potentially appropriate, but where a clear moral stance is still maintained. Some suggestions are outlined for how flexible forms of policy analysis may be used to combine an acceptance of the validity of a vast range of humanitarian actions while still retaining a strong stance against practices that may harm beneficiaries or feed the causes of conflict. Realism about each agency's room for manoeuvre is essential, especially local institutions. A modest but principled stance will involve helping actors to consider the impact of their work on conflict and to find ways to improve the quality of their interventions as perceived by beneficiaries. PMID- 10379097 TI - US government natural disaster assistance: historical analysis and a proposal for the future. AB - Governments often provide grants or low-interest loans to disaster victims. Yet these programmes have proven to be quite costly. In addition, questions have been raised about associated behavioural incentives. Conceptually, government disaster insurance programmes should be more efficient, consistent and equitable than ex post facto disaster relief in the form of grants and loans. Yet the performance of government disaster insurance programmes has been mixed, at best. This article reviews the history of US federal natural disaster assistance to individuals and concludes with a recommendation for a new government role in the provision of disaster insurance. PMID- 10379098 TI - Race, ethnicity and disasters in the United States: a review of the literature. AB - In this paper we synthesise past disaster research that addresses issues of race and ethnicity in the United States. Using an eight-stage typology to organise the findings, this literature review presents the results from a wide range of studies. The synthesis shows how various racial and ethnic groups perceive natural hazard risks and respond to warnings, how groups may be differentially affected, both physically and psychologically, and how disaster effects vary by race and ethnicity during the periods of emergency response, recovery and reconstruction. We show that studies have important findings, many illustrating that racial and ethnic communities in the US are more vulnerable to natural disasters, due to factors such as language, housing patterns, building construction, community isolation and cultural insensitivities. By presenting these studies together, we are able to witness patterns of racial and ethnic inequalities that may be more difficult to see or interpret in individual studies that take place in one specific time and place. We conclude the review with policy and research recommendations. PMID- 10379099 TI - Community-based disaster management during the 1997 Red River Flood in Canada. AB - This paper examines the relationship between community preparedness and response to natural disaster and their level and pattern of community development. This is done by investigating preparation and response to the 1997 Red River Flood by three rural communities in Manitoba, Canada. The communities were selected because of their different ethnic mix and associated level and pattern of community development. The hypothesis was supported that the level and pattern of community development affect community capacity to respond to flooding. Communities characterised by higher levels of physical, human and social capital were better prepared and more effective responders to the flood. However, where the pattern of community development was characterised by high levels of social capital, decision-making processes were complicated. PMID- 10379100 TI - Life cycle, host restriction and longevity of Cyrilia nili (Haemogregarina nili Wenyon, 1909) n. comb. AB - Morphometric and morphological observations on C. nili in an experimentally infected fish (using the leech vector Batracobdeloides tricarinata for the first time) revealed two successive types of merogonic cycles occurring within the fish erythrocytes. In the first cycle large meronts produced eight small merozoites each while in the second cycle smaller meronts produced four merozoites each. Merozoites of the second cycle were destined to become gamonts. The gamonts were somewhat larger than merogonic stages and comprised the majority of the parasitic stages during parasitemia for up to 8 months. In the leech crop, gamonts released from the fish erythrocytes, became associated in syzygy and fused. The formed zygote underwent sporogony within the intestinal tissues and up to 60 sporozoites were produced from an ovoid or round oocyst. The produced sporozoites migrated toward the salivary and the proboscis tissues. The parasite survived in the leech over a period of 8 months involving 5 meals after the initial meal provided the fasting period did not exceed 75-105 days. Survival was attributed to residual stages in the proboscis and salivary tissues. Cross transmission experiments between fishes via B. tricarinata revealed that the fish haemogregarines were not host specific. Previously described African fresh water fish haemogregarines were indistinguishable and on the basis of the current results, they are regarded as a single species Cyrilia nili. PMID- 10379101 TI - [The diagnosis of swine dysentery and spirochaete diarrhea. 1. Cultural biochemical differentiation of intestinal Serpulina in routine diagnosis]. AB - Frequent incidence of Serpulina strains showing all cultural and biochemical characteristics of Serpulina (S.) hyodysenteriae except of being indole negative, and alpha-galactosidase positive isolates showing strong haemolysis on Columbia agar with 5% sheep blood and trypticase soy agar with 5% ox blood, respectively, was the cause to evaluate common biochemical and cultural methods in Serpulina routine diagnostics. To this purpose ten type and reference strains as well as 47 field strains were examined for their ability to produce indole, haemolysis, hippurate cleavage, alpha-galactosidase, alpha- and beta-glucosidase activity. Two four-hour identification-systems were used, RapID ANA II and Rosco diagnostic tablets. The ability to produce indole was determined by different methods. All investigations were carried out at least two times. For the investigation of haemolytic patterns trypticase soy agar with 10% ox blood proved to be most effective. Results received using this agar could always be confirmed by the ring phenomenon. Determining the ability to produce indole by adding p dimethylaminocinnamaldehyde to bacterial growth collected on a cotton swab was confirmed to be more sensitive than other methods. Both four-hour-systems were shown to be useful in Serpulina diagnostics, though in the RapID ANA II only four of 18 available reactions could be used and the hippurate cleavage reaction has to be carried out additionally. Using cultural and biochemical methods, it was possible to assign the type and reference strains to the correct species, as well as 46 of 47 field isolates could be identified including all five known intestinal Serpulina species from swine. 27 strains were determined as S. hyodysenteriae, nine of these isolates atypically being indole negative. In contrast one canine S. pilosicoli strain was atypical showing indole production. Therefore incidence of indole negative variants of S. hyodysenteriae as well as indole positive S. pilosicoli isolates must be taken into consideration. PMID- 10379102 TI - Serotypes and electrophoretic protein profiles of Pasteurella haemolytica isolated from pneumonic ovine lungs. AB - Serotypes and SDS-PAGE protein profiles of P. haemolytica isolated from pneumonic ovine lungs were investigated. Of 268 P. haemolytica isolates, 232 (86.6%) were serotypable. A total of 12 serotypes were recognized in 20 different geographic origins of central Turkey. The most common serotype was A2, followed by A7, A1 and T4. Serotypes A13, A14, A16 and T15 could not be detected. In SDS-PAGE, marked differences between major bands of biotype A and T strains were found. In numerical analysis of protein profiles, biotype A and T strains were separated at 58% similarity level. Biotype A isolates produced a cluster at 80% similarity level, and biotype T isolates at 92% similarity level. No single cut off level was able to discriminate between each serotype studied and isolates could not be clustered on the basis of their geographic origins. PMID- 10379103 TI - [Possibilities and limits for therapy of amniotic hydrops in cattle. A review]. AB - The review describes the occurrence, causes and clinical signs of bovine placental hydrops. Furthermore prognosis, operative and hormonal treatment possibilities for interruption of pathological gestation as well as subsequent treatment are discussed. PMID- 10379104 TI - [The human being from the point of view of animals or, "Who knows whether the breath of men goes upwards?" (Ecclesiastes 3:21)]. AB - The question of whether animals have souls has been asked since as early as the Old Testament. Where this is believed to be true, fiction has provided interesting models in literature: The human being as seen by animals has been a popular subject since Apuleius' 'Asinus aureus' and how man appears from the perspective of a donkey or a beetle, that is to say the perspective from below, becomes controversial. Examples may be found in all languages and centuries in Jonathan Swift, Miguel Cervantes, E. T. A. Hoffmann, Ludwig Tieck, Heinrich Heine, Viktor von Scheffel, Franz Kafka and others. Resume at the end: How does man answer this question or his own self-questioning? PMID- 10379105 TI - Maxillary sinus augmentation prior to placement of endosseous implants: A histomorphometric analysis. AB - The aim of this prospective study was to histomorphometrically evaluate at various time intervals the mineralization stage and process of an allogeneic xenogeneic bone graft used in sinus augmentation procedures. One biopsy was taken from 20 patients at either 6, 8, 10, or 12 months after sinus augmentation. Immediately following the biopsy, an endosseous implant was placed into the biopsy site. This protocol provided 4 groups of 5 patients each, based on healing time following sinus augmentation. Using backscattered electron image analysis, the specimens were histomorphometrically analyzed to determine the volume fractions of residual cancellous bone, newly formed bone, soft tissue, bovine hydroxyapatite, and "remineralized" freeze-dried demineralized bone allograft (rDFDBA). "Remineralization" of DFDBA particles was observed in a few areas in all specimens. Polarized light microscopy showed that only the 12-month biopsies had a predominance of lamellar bone formation. The area within the biopsies that represented the residual alveolar ridge consisted of 32.6% +/- 8.6% (mean +/- SD) of bone. In the grafted area of the biopsies the volume fraction of newly formed bone at 12 months (20.7% +/- 8.3%) was significantly higher (P < .05, analysis of variance) than at 6 months (8.1% +/- 3.0%). There was no statistically significant difference between newly formed bone in the inferior, central, and superior grafted areas in all 4 time intervals. This prospective study indicates that the mineralization process of an allogeneic-xenogeneic sinus graft is incomplete 6 months after the sinus augmentation procedure. New bone formation increased up to 12 months postaugmentation; however, it remained lower than the volume of residual bone. PMID- 10379106 TI - Numerical analysis of a dental implant system preloaded with a washer. AB - Gold screw loosening is a problem that frequently affects dental implants. The application of a preload has been the main means of preventing loosening. However, this measure has not been able to eliminate its occurrence. In this study the effect of a washer in a Branemark-type implant on the loosening conditions of the retaining screw was investigated using a finite element simulation. The simulation indicated that a washer may significantly increase the tolerance of a screw against loosening. This is accomplished by increasing the tolerance of the implant against deformation. The addition of a customized washer to a dental implant system may offer a very simple and inexpensive solution for the persistent problem of screw loosening. PMID- 10379107 TI - Histologic evaluation of the periodontium of abutment teeth in combination implant/tooth fixed partial denture. AB - The histologic response of the periodontal tissues of teeth rigidly joined to implants with a fixed partial denture was evaluated using light microscopy. The fourth premolar of a dog was connected to implants placed in the first and second premolar position with a fixed partial denture. The restored teeth were under function for periods of 6, 12, 18, and 24 months, with unrestored fourth premolars as controls. The histology of the periodontal ligament on the fourth premolar was found to be similar in the control and the restored teeth. The periodontal tissues contained a minimal amount of inflammatory cell infiltrate. The crestal bone was cortical in nature, showing no periodontal breakdown. The orientation of the periodontal fibers was easily determined, indicating that minimal remodeling had taken place. The number and morphology of the blood vessels were also similar in the control and the treated teeth. The lack of inflammation and stability of the periodontal tissue suggested that the use of combination implant-to-natural-teeth restorations with rigid joints in this animal model does not result in deleterious effects on the periodontal tissues and that the forces placed on the tissues are within the remodeling capabilities of the teeth. PMID- 10379108 TI - Evaluation of titanium implants placed into simulated extraction sockets: a study in dogs. AB - The purpose of this study was to evaluate the effect of gap width on bone healing around implants placed into simulated extraction socket defects of varying widths in 10 mongrel dogs. All premolars were removed and the alveolar ridges were reduced to a width of 7 mm. Nine weeks later, a total of 80 implants, 10 mm long by 3.3 mm wide, were placed into osteotomy sites prepared to 3 different diameters in the coronal half, simulating extraction sockets. Three experimental sites, with gap sizes of 0.5 mm, 1.0 mm, and 1.4 mm, were created; the control sites had no gap. The depth of each defect was measured at the time of implant placement. All implants were stable at the time of placement. The dogs were euthanized 12 weeks after implant placement, and blocks containing the implants and adjacent bone were submitted for histologic evaluation. Clinically, all control and test sites healed, with complete bone fill in the defect. Percentages of bone-to-implant contact were measured histologically. As the gap widened, the amount of bone-to-implant contact decreased, and the point of the highest bone-to implant contact shifted apically. These changes were statistically significant (P < .001). No statistically significant differences in bone-to-implant contact were found between the sites when the apical 4 mm of implants were compared. Within the limits of this study, the simulated extraction socket defects healed clinically, with complete bone fill, regardless of the initial gap size. However, the width of the gap at the time of implant placement had a significant impact on the histologic percentage and the height of bone-to-implant contact. PMID- 10379109 TI - Eighteen-month radiographic and histologic evaluation of sinus grafting with anorganic bovine bone in the chimpanzee. AB - Maxillary sinus grafting procedures are currently the treatment of choice when the alveolar crest of the posterior maxilla is in close approximation to the maxillary sinus. The short-term histologic and radiographic healing following sinus grafting with natural bone mineral (Bio-Oss) in the chimpanzee has been evaluated. We have previously shown by histomorphometric and radiographic analysis that the percentage of vital bone area, the vertical height, and the density of new bone in the maxillary sinus was significantly greater with anorganic bovine bone compared to bovine Type I collagen matrix. The purpose of this in vivo study was to determine the bone mineral density (BMD) of the sinus grafts, the vertical height stability, the vital bone area, and the extent of anorganic bovine bone replacement 18 months postoperatively in 4 maxillary sinuses from 4 different animals. Radiographic analysis of computed tomographic scans taken at 1.5 years revealed an average BMD of 658 mg/mL, which was not significantly different from the values found at 6.5 months. The radiographic vertical height was maintained between the 6.5- and 18-month time points. On average, the grafts were found to have a height of 14 mm. Lateral wall biopsy specimens at 7.5 months were compared to those at 18 months. With the anorganic bovine bone treatment, the percentage of vital bone area increased from 62 +/- 3% to 70 +/- 7% and the percentage of natural bone mineral area decreased from 19 +/ 14% to 6 +/- 3%. The bovine Type I collagen matrix vital bone percentage at 7.5 months was 34 +/- 21%. These results demonstrate that sinus grafting with anorganic bovine bone maintains radiographic evidence of density and height stability of 1.5 years. In addition, histologic evidence supports the hypothesis that anorganic bovine bone is replaced by vital bone. PMID- 10379110 TI - Marginal discrepancy of screw-retained and cemented metal-ceramic crowns on implants abutments. AB - This in vitro study quantified the marginal discrepancy of the implant-to prosthetic-crown interface on nonsubmerged dental implants restored with either a cemented or a screw-retained approach. Metal-ceramic crowns were fabricated for 20 ITI 4.1 x 10 mm solid-screw titanium implants. Ten implants received octa abutments and screw-retained crowns fabricated on premachined gold cylinders. The remaining 10 implants were restored with 5.5-mm solid abutments and metal-ceramic crowns cemented alternately with a glass-ionomer or a zinc phosphate luting agent. Inspection of the implant-crown interface was conducted using light microscopy under x 50 magnification at selected stages in the process of crown fabrication. Statistical analysis revealed a significant difference (P < .001) in the mean marginal fit between screw-retained (8.5 +/- 5.7 microns) and luted implant-supported crowns. This difference was observed both before (54.4 +/- 18.1 microns) and after cementation with glass-ionomer (57.4 +/- 20.2 microns) or zinc phosphate (67.4 +/- 15.9 microns). PMID- 10379111 TI - Assessment of the blood supply to the mental region for reduction of bleeding complications during implant surgery in the interforaminal region. AB - Several life-threatening complications caused by hemorrhage that can occur during the placement of dental implants in the mandibular interforaminal region have been described in the literature. The aim of this study was to assess the vascular supply to this region and delineate the relative contribution and importance of the sublingual artery versus the submental artery. Thirty-four human cadavers were dissected. Special attention was directed to the presence of a branch of the submental artery that perforates the mylohyoid muscle and thus participates in the blood supply to the floor of the mouth together with the sublingual artery. A sublingual artery was found in 71% of the specimens. A large branch of the submental artery perforating the mylohyoid muscle was found in 41% of the specimens. The point of perforation was located an average of 31 mm posterior to the menton. The high risk of injuring the vessels of the floor of the mouth can be explained by the close vicinity of these vessels to the mandibular lingual cortical plate. To prevent complications in cases of unclear anatomic identification of the fossa sublingualis, preoperative lingual probing or elevation of the periosteum of the lingual aspect of the mandible is necessary. An alternative diagnostic procedure is precise preoperative noninvasive imaging (eg, computed tomography). PMID- 10379112 TI - Osseointegration of rough acid-etched titanium implants: 5-year follow-up of 100 minimatic implants. AB - During 1992, 100 Minimatic screw implants made of titanium alloy (titanium aluminum-vanadium) with a machined rough acid-etched surface were placed in 63 consecutive partially edentulous patients. At second-stage surgery, which was performed after a 4- to 6-month healing period, none of the implants showed signs of mobility, peri-implant infection, or bone loss from the crest of the ridge. Each patient was restored with a fixed prosthesis and reexamined every 3 months during the first year. Periapical radiographs were taken annually up to 5 years. These revealed no signs of peri-implant radiolucencies involving any of the implants, and mean alveolar bone loss was less than 1 mm at the 5-year examination. One implant was considered a late failure because of a peri-implant infection that developed during the first year, although the implant was still functional at year 5. Another patient with 2 implants dropped out during the fifth year of the study, although both implants had been considered successful up to that point. Based on annual measurements of Plaque Index, Sulcular Bleeding index, pocket probing depth, attachment level, width of keratinized mucosa, and hand-tested mobility, 97 of the remaining 98 implants were considered successful, resulting in a 98% success rate. This 5-year study confirms that Minimatic machined acid-etched implants provide predictable osseointegration results and supports the conclusion of other reports that titanium implants with a rough surface can fulfill the requirements of Albrektsson et al (1986) for implant success. PMID- 10379113 TI - Maintenance of regenerated bone beneath pontics: preliminary clinical report of 43 sites. AB - Ridge augmentation was achieved through the use of guided bone regeneration procedures in pontic areas of 43 planned fixed prostheses. Measurements taken through templates, which fit over the final fixed prostheses, at the time of prosthetic placement and a mean of 123 weeks after prosthesis placement demonstrated a change of less than 0.1 mm in buccopalatal dimensions of the regenerated hard tissues. PMID- 10379114 TI - Analysis of 356 pterygomaxillary implants in edentulous arches for fixed prosthesis anchorage. AB - One thousand eight hundred seventeen implants were placed in the completely edentulous maxillae of 189 patients (122 female; 67 male). The patients' mean age was 60 years (range 28 to 91 years). Three hundred fifty-six of the 1,817 implants were placed in the pterygomaxillary area, and all patients were restored with complete-arch fixed detachable prostheses. The mean number of implants per maxillary prosthesis was 9.0 (range 6 to 15). During stage II surgery and before loading, 41 pterygomaxillary implants (11.5%) were not osseointegrated and were removed. After a mean loading period of 4.68 years (range 0.06 to 9.20 years), 1 additional pterygomaxillary implant was lost. Altogether, 42 of 356 pterygomaxillary implants (11.8%) were removed. Survival rates according to implant size, bone quality, and tooth position were also recorded. This study illustrates a cumulative survival rate of 88.2% for pterygomaxillary site implant placement in edentulous maxillary arches. PMID- 10379115 TI - Histomorphometric evaluation of extraction sockets and deficient alveolar ridges treated with allograft and barrier membrane: a pilot study. AB - The aim of the study was to determine the fate of demineralized freeze-dried bone allograft (DFDBA) used in conjunction with a barrier membrane in the management of extraction sockets and deficient alveolar ridges, and to compare the amount of bone formed with that found in untreated sites. Ten biopsies were obtained from 8 grafted patients. Five biopsies were harvested from untreated sites during routine implant placement and analyzed for comparison. In the socket management procedure, DFDBA was packed tightly into the socket and covered with an expanded polytetrafluoroethylene (e-PTFE) membrane. Primary closure was achieved in all cases. In the ridge regeneration procedure, cortical columns were placed in the ridge projecting outward approximately 3 mm to create and maintain space for DFDBA particles packed between them; the columns were then covered by an e-PTFE membrane. Healing time ranged from 8 to 23 months. At the time of implant placement, bone cores (7 mm x 2 mm) were harvested, fixed in 10% formalin solution, and prepared for histologic examination. At the light microscopic level, no inflammation or fibrous encapsulation was observed. New bone formation on and around DFDBA particles was widespread. Histomorphometric analysis of the grafted specimens and untreated sites was carried out using the trabecular bone volume (TBV) index. The TBV in the maxillary test specimens was 55.03%, as compared to 57.33% of control cores. Unaltered DFDBA made up 8.7% of the test specimens. In the mandibular biopsies, the TBV was 56.6%, while for the controls it was 40.9%. The volume of DFDBA still present was 2.45%. The results tended to indicate that treatment with DFDBA in conjunction with cell occlusive membranes will result in new bone formation, predominantly by the process of conduction, which appears to be similar in amount and nature to that found in cores harvested from healed nonfunctional edentulous areas. PMID- 10379116 TI - Implant-surgical and prosthetic rehabilitation of patients with multiple dental aplasia: a clinical report. AB - The expanded experience with oral implants and supplementary augmentation techniques has opened new possibilities for treating patients with oligodontia or anodontia with fixed prostheses. A problem in treating such patients is the need to place implants in growing maxillae or mandibles, as many of these patients are children or adolescents. When implant treatment is postponed until the patient is full grown, dysfunctions become manifest, which necessitates extensive surgical measures to achieve a fixed prosthetic restoration. This report illustrates the problems associated with different concepts for the treatment of multiple aplasia with implants. The results are based on the findings of 22 patients with oligodontia who underwent surgical treatment and were followed over a period of 5 years. Two controversially treated cases are presented. PMID- 10379117 TI - Use of crestal bone for augmentation of extremely knife-edged alveolar ridges prior to implant placement: report of 3 cases. AB - A technique is presented for interforaminal lateral augmentation of mandibles with adequate bone height, but extremely knife-edged mandibular alveolar ridges (Class IV of Cawood and Howell's classification of residual ridges), in which the crestal portion of the knife-edged ridge is used as grafting material. Following an osteotomy and rotation of the grafts by 180 degrees, the grafts were fixed to the residual ridge below the osteotomy line by means of miniscrews. All grafts showed only mild resorption after a healing period of 3 months, and it was possible to place 4 implants in the now sufficiently wide host region. PMID- 10379119 TI - Soft tissue exposure of endosseous implants between stage I and stage II surgery as a potential indicator of early crestal bone loss. AB - Implants are well accepted as a means of dental rehabilitation. While integration success rates are high, crestal bone loss can occur, and it may not become apparent until stage II surgery and implant uncovering. The purpose of this study was to quantify the relationship between exposure of implants through the oral mucosa between stage I and stage II implant surgery and early changes in crestal bone height. Bone levels were measured during placement of 275 implants in the maxillae of 50 subjects. Repeated bone height measurements were obtained at implant uncovering. Fourteen implants in 7 patients were exposed to the oral cavity through the mucosa at stage II surgery. Patients with 1 or more exposed sites demonstrated a likelihood of bone loss 3.9 times greater than patients with nonexposed sites (Fisher exact test, P = .0003). These results suggest that exposure of an implant between stage I and stage II implant surgery might serve as a potential indicator of the occurrence of early bone loss. PMID- 10379118 TI - Longitudinal evaluation of aspartate aminotransferase in the crevicular fluid of implants with bone loss and signs of progressive disease. AB - Aspartate aminotransferase (AST) has been shown to be a promising host marker for periodontal disease progression. The aim of the present study was to analyze AST in the crevicular fluid (CF) of implants exhibiting peri-implantitis and to evaluate the association between AST levels and progressive attachment loss. Twenty patients who had received a total of 42 endosseous cylindric titanium implants were examined. Radiographic assessment of preexisting bone loss and clinical measurements, including electronic attachment of probing, presence or absence of plaque, bleeding on probing, and AST analysis in CF, were performed on 2 occasions 6 months apart. During this study period 13 of 168 sites in 7 patients experienced further loss of attachment greater than or equal to 1.0 mm (median 1.7 mm; interquartile range 0.4 mm). Evaluation of a positive AST test (> or = 300 microIU) in site-specific diagnosis revealed low positive (8%) and high negative predictive values (92%), with a sensitivity of 15% and a specificity of 83%. These results indicate that, in contrast to periodontal disease, the assessment of AST in peri-implant crevicular fluid may be of limited value as a diagnostic and prognostic marker for peri-implant disease. PMID- 10379120 TI - Histologic evaluation of clinically successful osseointegrated implants retrieved from irradiated bone: a report of 2 patients. AB - The present study described the histologic findings of 2 implants and surrounding tissues retrieved from human irradiated bone. For the treatment of a malignant tumor, 50 Gy of irradiation after implant placement and 60 Gy of irradiation before implant placement were provided for patients 1 and 2, respectively. In patient 1, the implant and surrounding tissues were removed from the frontal bone 24 months after implant placement because of the patient's death from a tumor recurrence. In patient 2, the implant and surrounding tissue were removed from a maxillectomy site 26 months after implant placement because of tumor recurrence. In each patient, new bone formation surrounding the implants was observed. The ratio of direct bone-implant contact along the threaded implant surface was 61.3% in patient 1 and 69.0% in patient 2. The ratio of the area occupied by mineralized bone in each thread was 75.8% in patient 1 and 81.2% in patient 2. These results indicate the potential of irradiated bone to achieve osseointegration of titanium implants. PMID- 10379121 TI - Obstetrics and gynaecology in the Asia and Oceania region: requirements for postgraduate training and certification. AB - This paper reviews the requirements of postgraduate training and certification in the countries of the Asia and Oceania regions. It explores the feasibility of establishing exchange training programmes, and presents recommendations which might facilitate national societies in standardising their programmes. PMID- 10379122 TI - The effect of epidermal growth factor on the preimplantation development, implantation and its receptor expression in mouse embryos. AB - OBJECTIVE: To investigate the influence of epidermal growth factor (EGF) on preimplantation development, implantation, and expression of epidermal growth factor receptor (EGFR) itself in mouse embryos. MATERIALS AND METHOD: Eight-cell stage mouse embryos were cultured for 48 hours with EGF at concentrations of 0.1, 1.0, 10 and 100 ng/ml. Embryos not treated with EGF were served as control. The percentages of embryos which developed to the expanded, hatched blastocyst stage and in vitro implantation at 48 hours were determined. Reverse transcription polymerase chain reaction (RT-PCR) has been used to examine the expression of EGFR in developed hatched blastocysts. Following reverse transcription, strategically designed nested primers, optimized for specificity, were used for amplification from the cDNA equivalent of a single embryo. The products were then verified by restriction enzyme digestion and sequence analysis. Results were analyzed with chi 2 test and Student's t-test as appropriate, and statistical significance was defined as p < 0.05. RESULTS: The percentages of fully expanded blastocysts at 48 hours in all the EGF treated group were not significantly different from the control. The percentages of hatched blastocysts were significantly higher in the EGF treatment group at 0.1 ng/ml (90.5 +/- 9.8%) compared to the control (82.1 +/- 7.2%), 1.0 ng/ml (82.2 +/- 12.7%), and 100 mg/ml (81.9 +/- 11.8%) (p < 0.05, p < 0.05, p < 0.05, respectively). The percentages of hatched blastocysts were significantly higher in the EGF treatment group at 10 ng/ml (89.4 +/- 7.5%) compared to the control, and 100 ng/ml (p < 0.05, p < 0.05, respectively). The percentages of attached blastocysts in vitro were significantly higher following incubation with EGF at concentrations of 0.1 ng/ml (37.0 +/- 17.0%), 1.0 ng/ml (32.0 +/- 14.3%), 10 ng/ml (21.3 +/- 7.2%) compared to the control (9.5 +/- 7.7%) (p < 0.05, p < 0.05, p < 0.05, respectively). The attachment rates in 0.1 ng/ml and 1.0 ng/ml EGF treatment groups were also significantly higher than those in other EGF treatment groups. Embryo development and attachment were not significantly inhibited or enhanced in cultures supplemented with 100 ng/ml EGF compared to the control. The mRNA concentration of EGFR in embryos treated with 0.1 ng/ml of EGF was significantly higher than those of the control and other EGF treatment groups. CONCLUSION: EGF may have a stimulatory role in later stage embryonic development, implantation and expression of EGFR in hatched blastocyst itself at the specific concentration. PMID- 10379123 TI - Umbilical Doppler velocimetry prediction of discordant twins. AB - OBJECTIVE: To evaluate the role of umbilical Doppler velocimetry as a comprehensive test for the prediction of discordant twins. METHODS: The sets of twins were studied with duplex Doppler velocimetry for umbilical artery in third trimester. A systolic/diastolic ratio was measured for each twin. The average difference in the ratios between each twin > or = 0.4 was used to indicate abnormal test. Discordancy was identified when the birth weight difference of > 25%. RESULTS: Among the 52 sets of twin pregnancies studied, 40 sets of twins fulfilled the study criteria. Eight sets of twins were discordant (20%). The mean gestational age at delivery was 37.15 +/- 2.24 weeks (range 28 to 41 weeks). The test correctly identified 6 of the 8 growth discordant twins which had a sensitivity of 75%, specificity of 68.75%, and accuracy of 70%. CONCLUSION: Umbilical Doppler velocimetry is useful in prediction of discordant twins. PMID- 10379124 TI - Assessment of myometrial invasion at the invasion site of an endometrial carcinoma by ultrasonography along with an intrauterine catheter. AB - OBJECTIVES: To evaluate by transvaginal ultrasonography (TVU) the thickness of the intact myometrium at the presumed tumor-origin site and to establish criteria for a half myometrial invasion. METHODS: A total of 19 successive patients with endometrial cancer who were treated between January 1, 1997, and January 31, 1998, participated in this study. TVU mode B with and without the use of an intrauterine silicon catheter was performed. RESULTS: Using a catheter, the origin site was correctly detected in 15 cases (79%). The best criterion for half myometrial invasion was a 6-mm thickness of the intact myometrium at the origin site. The sensitivity/specificity/accuracy of TVU with the use of a catheter in cases with the correct estimated origin site, were 1.00/0.67/0.86 for myometrial invasion < 1/2, and 0.67/1.00/0.86 for myometrial invasion > or = 1/2. CONCLUSION: It is of value to use 6-mm as the criterion for the thickness of the intact myometrium at the estimated tumor-origin site in connection with TVU with the use of a catheter for preoperatively assessing half myometrial invasion. PMID- 10379125 TI - A randomized controlled trial comparing midwife-managed care and obstetrician managed care for women assessed to be at low risk in the initial intrapartum period. AB - OBJECTIVE: To compare the efficacy of midwife-managed care and obstetrician managed care for women assessed to be at low risk in the initial intrapartum period. METHODS: 1,050 women assessed to be at low risk on admission to labour ward in the Prince of Wales Hospital participated in this study. By computer generated random allocation, 563 (54%) women were assigned to Group A (experimental) under midwifery care, and 487 (46%) women to Group B (control) under obstetrician care. The outcomes and complications between the 2 groups were compared. Data were analyzed by 2 x 2 contingency tables and Chi-square. RESULTS: 150 (26.6%) women in the experimental group were taken over by the obstetricians. 46 (30.7%) women were transferred to obstetrician-management for the preference of epidural analgesia. The other reasons for taken over the remaining 104 (69.3%) women were fetal distress, poor progress of labour, complications in first or second stage of labour. The experimental group had less oxytocic augmentation (Chi-square = 7.49, p = 0.006) and the insertion of intravenous infusion (Chi square = 5.34, p = 0.02). Both groups had similar outcomes on normal delivery, operative vaginal delivery, caesarean section and complications. CONCLUSIONS: Midwife-managed care is as safe as obstetrician-managed care for women who were assessed to be at low risk in the intrapartum period. Routine visit by obstetrician is not necessary and the midwives are able to detect complications in the course of labour and alert the obstetrician for taking the necessary action. PMID- 10379126 TI - A case of simultaneous presence of primary endometrial carcinoma and metastasis of a breast carcinoma to the ovary after 5 years of tamoxifen therapy. AB - We report a case of a 43-year-old woman with the simultaneous presence of a primary uterine endometrial cancer and metastasis of breast cancer to the ovary after 5 years of tamoxifen therapy. Tamoxifen therapy lengthens recurrence-free survival of the patient. However, the risk of endometrial cancer and the possibility of recurrence of breast cancer also must be considered. PMID- 10379128 TI - Primary malignant melanoma of the uterine cervix: a case report. AB - We report on a case of a malignant melanoma of the uterine cervix. Histological and immunohistological examinations of a postsurgical specimen revealed malignant melanoma. The junctional activities did not occur due to extensive superficial ulceration. Radical surgery was performed. The patient is doing well and free of symptoms at this time, 2 1/2 year later. PMID- 10379127 TI - A prospective randomized study comparing hysteroscopy and curettage (H & C) under local anaesthesia (LA) and general anaesthesia (GA) in Chinese population. AB - OBJECTIVE: To compare the difference in patient's acceptance of local anaesthesia (LA) and general anaesthesia (GA) hysteroscopy and curettage in Chinese population. DESIGN: A prospective randomized study. SUBJECTS AND METHODS: In the period September 1994 to August 1995, all Chinese women with abnormal uterine bleeding or suspected uterine anomaly who warranted hysteroscopy and uterine curettage were invited to participate in this study with informed consent. They were randomly allocated to the control (i.e. GA) and study (i.e. LA) group. RESULTS: Overall 90% of the controls and 91% of the study group were satisfied with the procedure. The hysteroscopic diagnostic accuracy was 83%. Significantly higher percentage of patients in the study group opted for the same form of admission arrangement if given the choice. CONCLUSION: Hysteroscopy and curettage under LA and GA are equally acceptable in the Chinese population in Hong Kong. The patient satisfaction rate is high in both groups. Hysteroscopic diagnosis is highly accurate in malignant condition (100% sensitivity and 83% specificity). PMID- 10379129 TI - Prenatal diagnosis of intrapericardial teratoma: a case report. AB - A hydropic fetus that exhibited intrapericardial teratoma with marked pericardial effusion was prenatally diagnosed. Intrauterine pericardiocentesis was performed to diagnose and to treat hydrops. Pericardiocentesis for tamponade secondary to a fetal intrapericardial teratoma might prevent fetal death and premature delivery. PMID- 10379130 TI - Previous cesarean section and abortion as risk factors for developing placenta previa. AB - OBJECTIVE: To determine the risk of subsequent occurrence of placenta previa in women with a history of previous cesarean sections and/or spontaneous and induced abortions. METHODS: A retrospective analysis of all single gestation deliveries at National University Hospital of Singapore from 1993-1997 was done. Women with placenta previa were identified by clinical or ultrasonographic diagnosis. RESULTS: Of the 16,169 singleton deliveries, 164 women (1.0%) had placenta previa. Women with placenta previa had a significantly higher incidence of previous cesarean sections (p < 0.001). Among the 164 women with placenta previa, women with 1, 2, and 3 previous cesarean sections had 2.2 (95% CI 1.4, 3.4), 4.1 (95% CI 1.9, 8.8) and 22.4 (95% CI 6.4, 78.3) times increased risk of developing placenta previa respectively. Similarly, women with 2 or more previous abortions had a 2.1 (95% CI 1.2, 3.5) times increased risk of subsequently developing placenta previa. CONCLUSION: There is a strong association between previous cesarean section and risk of subsequent development of placenta previa. This risk increased with the number of previous cesarean sections. Increasing frequency of abortions was also found to predispose a woman to placenta previa. PMID- 10379131 TI - Thermal balloon endometrial ablator: a preclinical safety and effectiveness study. AB - OBJECTIVE: To evaluate the preclinical safety and efficacy of the thermal balloon endometrial ablator (TBEA). METHODS: Informed consent was taken from all patients for TBEA and hysterectomy at same sitting. TBEA was performed in vivo just before hysterectomy on 6 patients' uteri to determine uterine rupture, perforation and thermal damage to surrounding viscera and peritoneum. Temperatures in the pouch of Douglas, uterovesical peritoneum and serosa of uterus were taken by a sterile laboratory thermometer to note for any rise in temperature before, during and after the procedure. The extirpated uteri were then examined grossly for the nature and extent of thermal damage which was visible as a zone of erythema in the endomyometrium. Five extirpated uteri, 3 from the above group and 2 on whom TBEA was performed 3 months ago were histopathologically examined to study the nature and extent of damage to endomyometrial cells. A 5 mm longitudinal strip of uterus along with both the cornual ends were sectioned in such a way that it was representative of the entire uterus. RESULTS: There was no evidence of uterine perforation, rupture or any damage to the surrounding viscera and peritoneum. There was no rise in temperature in most sites while TBEA was performed. The only area that felt warm was the surface of the uterus and there too the measured rise was only 1 degree C. On gross examination the zone of erythema measured 5.4 mm (mean) (range 3-9 mm). In those uteri subjected to histopathology immediately after TBEA early evidence of thermal damage was visible as hemorrhage, congestion, edema, eosinophilic infiltration and necrosis in 2 out of 3 cases. Late changes of thermal damage after TBEA were seen as basal endometrium and areas of hyalinization. The changes in general were patchy and not uniform. CONCLUSIONS: TBEA is a safe device with no thermal damage to uterus and surrounding viscera. The mean zone of thermal damage in the endomyometrium is 5.4 mm. On histopathology, the early changes of thermal damage are hemorrhage, congestion, edema, eosinophilic infiltration and necrosis and the late changes are visible as areas of hyalinization and presence of basal endometrium. PMID- 10379132 TI - Foley's catheter for cervical ripening. PMID- 10379133 TI - Empirically supported treatments in pediatric psychology: severe feeding problems. AB - OBJECTIVE: To identify treatment studies for severe pediatric feeding problems that meet the modified methodological criteria of the Task Force on Promotion and Dissemination of Psychological Procedures (1995). METHODS: Articles in peer reviewed medical and psychological journals (1970-1997) reporting psychosocial or behavioral intervention studies targeting an identified oral feeding problem in children were selected. Methodologically rigorous studies were identified and treatments were classified as well established, probably efficacious, or promising interventions according to specified criteria. RESULTS: Effective interventions for children with severe feeding problems are contingency management treatments that include positive reinforcement of appropriate feeding responses and ignoring or guiding inappropriate responses. Promising interventions include positive reinforcement for acceptance and not removing the spoon for refusal and swallow induction training. CONCLUSIONS: Because only studies of behavioral interventions met methodological criteria, well-controlled intervention studies are needed across a variety of theoretical perspectives. Empirically supported treatments for feeding problems exist; it is now time to turn to questions about for whom they are appropriate, and when, and why. PMID- 10379135 TI - Commentary: feeding problems: an ecological perspective. PMID- 10379136 TI - Commentary: beyond feeding problems: the challenge of meeting dietary recommendations in the treatment of chronic diseases in pediatrics. PMID- 10379137 TI - Empirically supported treatments in pediatric psychology: pediatric obesity. AB - OBJECTIVE: To review the efficacy of existing interventions for pediatric obesity with reference to the Chambless criteria. METHODS: Chambless criteria for determining treatment efficacy were applied to 42 randomized studies involving nonschool-based programs targeting childhood and adolescent weight loss. RESULTS: We summarize the following dimensions of the pediatric obesity treatment literature: description of participants, diagnostic criteria for study participation, experimental design, treatment protocol, treatment outcome, and follow-up. CONCLUSIONS: There is strong evidence for the short- and long-term efficacy of multicomponential behavioral treatment for decreasing weight among children relative to both placebo and education-only treatments. Conclusions about adolescent obesity treatment programs are more tentative as they have been less frequently examined, less rigorously controlled, and usually have not conducted long-term follow-up. Current research appears to be working to identify more efficacious treatments for pediatric obesity by exploring the specific behavioral strategies that will be most effective in modifying children's eating and physical activity habits. PMID- 10379138 TI - Commentary: empirically supported treatments for pediatric obesity: goals, outcome criteria, and the societal context. PMID- 10379139 TI - Commentary: future research directions in pediatric obesity research. PMID- 10379140 TI - Who participates in research on adherence to treatment in insulin-dependent diabetes mellitus? Implications and recommendations for research. AB - OBJECTIVE: Examine the implications of nonparticipation in studies of treatment adherence among adolescents with chronic health conditions. METHODS: Empirical data from an adherence study with adolescents with diabetes were used to demonstrate the influence of family participation on demographic and health outcome variables. Ninety-four families were categorized into one of three groups: (1) families that declined to participate in the study at recruitment (nonconsenters), (2) families that agreed to participate, but failed to return the study questionnaires (nonreturners), and (3) families that had at least one family member return the questionnaires (participants). RESULTS: Despite being similar demographically, nonreturners had significantly lower treatment adherence scores and the adolescents tested their blood sugar less frequently than participants. Participants and non-consenters did not differ on any available data. CONCLUSIONS: We discuss the implications of these group differences on the generalizability of research findings, offer suggestions about how to maximize and maintain participation in research studies, and suggest directions for future research. PMID- 10379141 TI - An experimental examination of learned helplessness in older adolescents and young adults with long-standing asthma. AB - OBJECTIVE: To examine the effects of experimentally induced learned helplessness in older adolescents and young adults with long-standing asthma. METHODS: Thirty nine participants (18-24 years of age) with histories of long-standing asthma (AS) and an age-matched healthy cohort (HC) (N = 94) received either contingent or noncontingent feedback on an experimental task. Participants' anagram-solving performance was assessed following the experimental procedure. Participants also completed a measure of depression and pretest-posttest measures of mood, expectancy, and attributions related to experimental task performance. RESULTS: The AS participants demonstrated significantly greater problem-solving deficits following response-noncontingent feedback, compared to the HC group. Further, whereas both AS and HC participants made more internal performance attributions when given response-contingent feedback, only AS participants demonstrated a pattern of increased internal attributions (i.e., self-focus) following response noncontingent failure. In addition, 21% of AS participants met DSM-IV criteria for major depression, compared to only 5% of the HC group. CONCLUSIONS: Individuals with long-standing asthma may be at increased risk for depression and for learned helplessness deficits, specifically impaired problem solving, in response to environmental noncontingency. Results are discussed in terms of both learned helplessness theory and perseverative self-focus conceptualizations of depression. The implications for both short- and long-term management of pediatric asthma are also discussed. PMID- 10379142 TI - Functional disability in adolescents and young adults with symptoms of irritable bowel syndrome: the role of academic, social, and athletic competence. AB - OBJECTIVE: To examine perceived academic, social, and athletic competence as potential moderators of the relation between symptoms of irritable bowel syndrome (IBS) and functional disability in adolescents and young adults with a history of recurrent abdominal pain (RAP). METHODS: We assessed IBS symptoms, competence, and disability by telephone interview in RAP patients five years following their medical evaluation. RESULTS: For both male and female subjects, the relation between symptoms and disability was stronger at lower levels of perceived academic competence. Furthermore, among females, the relation between symptoms and disability was stronger at lower levels of perceived social competence; among males, the relation was stronger at lower levels of perceived athletic competence. CONCLUSIONS: Perceived competence moderated the relation between IBS symptoms and functional disability. Interventions designed to enhance patient competence in various roles may be useful in reducing disability among adolescents and young adults with symptoms of IBS. PMID- 10379143 TI - Review: emotional and behavioral functioning in phenylketonuria. AB - OBJECTIVE: To examine 17 studies of the psychological sequelae of early-treated phenylketonuria (PKU) with emphasis on the impact of dietary control on functioning. Two questions are addressed: (1) What is the typical psychological profile associated with PKU? (2) Is emotional and behavioral disturbance more prevalent in PKU-affected individuals compared to appropriate controls? METHOD: Computerized searches of PsycINFO identified studies using behavioral, personality, and diagnostic measures. RESULTS: Findings converge upon a profile including attentional difficulties, depression, anxiety, and low self-esteem. Methodological constraints limit conclusions regarding the nature and severity of observed difficulties. A single study has used comparison groups appropriate for the simultaneous examination of the questions posed (Waisbren and Levy, 1991). CONCLUSIONS: We discuss results using a biopsychosocial framework, addressing the factors and processes that may influence emotional and behavioral functioning in this neurodevelopmental disorder. We outline potential lines of new investigation that address critical methodological factors. PMID- 10379144 TI - [Transvaginal ultrasonic evaluation of the thickness of the section of the uterine wall in previous cesarean sections]. AB - BACKGROUND: The aim of this study is to evaluate accuracy of transvaginal sonographic examination of the lower uterine segment in pregnant women with previous cesarean section. METHODS: Sixty-one pregnant women between 37 and 40 weeks of gestation, with previous cesarean section underwent transvaginal ultrasonography. Wall thickness of the lower uterine segment, the length of cervix, dilation of the isthmus uteri were measured. On the basis of the surgical findings (in 53 patients) and outcome of the trial of labor (in 8 patients) a Score was assigned to the pregnant women: Score 1 to the women who had good healing or a trial of labor without complications; Score 2 to the women with a thin or discontinued scar and in case of threatened rupture of the uterus in the trial of labor. RESULTS: The mean thickness of the lower uterine segment is 3.82 mm +/- 0.99 mm. The Score 1 group shows a mean thickness of 4.2 mm +/- 2.5 mm, and the Score 2 group a mean thickness of 2.8 mm +/- 1.06 mm. The transvaginal sonographic examination provides a sensitivity and a specificity respectively of 100 and 75%, for a thickness cut-off of 3.5 mm, and a positive and negative predictive values of 60.7% and 100% respectively. CONCLUSIONS: The transvaginal sonographic evaluation of the lower uterine segment improves therefore the obstetrical decision-making regarding the trial of labor in women with previous cesarean section. PMID- 10379145 TI - [Importance of endocervical cells in the diagnosis of CIN]. AB - BACKGROUND AND AIM: The authors performed a comparative study to define the role played by the presence of endocervical cells on the smear in the correct diagnosis of CIN. METHODS: The study was performed from January to December 1996 at the Clinic of Obstetrics and Gynecology of the Second University of Naples and involved 67 women with a histological diagnosis of CIN made in the two previous months of the study. The smears taken earlier were re-examined to assess the endocervical component regarding columnar and metaplastic cells and a comparison was made between smears which were CIN-positive and negative. RESULTS: The difference between positive and negative CIN smears was statistically non significant for columnar cells (66% vs 56%), unlike the findings for metaplastic cells (82% vs 61%). This demonstrated that CIN smears are more likely to include metaplastic cells compared to negative smears and the two types of smear do not differ significantly with regard to columnar cells. CONCLUSIONS: In order to make a cytological diagnosis of CIN, attention must predominantly be focused on the metaplastic component of endocervical cells. PMID- 10379146 TI - [Maternal-fetal transmission of HCV. Role of HIV as a risk factor]. AB - BACKGROUND: The aim of this study is to determine the rate of vertical transmission of hepatitis C and to analyse the concomitant infection by HIV as a risk factor. METHODS: We have studied the perinatal transmission of HCV in 22 pregnancies: 14 in women HCV+/HIV-, 8 in women HCV+/HIV+. We have performed the following tests on sera: test RIBA II to search for Ab anti-HCV, alanine transaminase (ALT) evaluation and HCV-RNA research by PCR. These tests were performed on sera from infants at birth and, then, during one year every three months. RESULTS: Within one year Ab anti-HCV disappeared in 20 of 22 pregnancies: two infants positive by Ab anti-HCV were born to HIV+ mothers and they were the only two who showed abnormal ALT values and detectable levels of HCV-RNA. Finally 10 of 14 infants born to HCV+/HIV- mothers were breast-fed and none was infected. CONCLUSIONS: We conclude that HCV mother-to-child transmission is an uncommon event, breast-milking is safety, and the concomitant infection by HIV could represent a risk factor for vertical transmission of hepatitis C. PMID- 10379147 TI - [Clinical characteristics of HCV infection in pregnancy: the role of sexual transmission]. AB - BACKGROUND: There is no uniformity of opinions about the possibility of sexual transmission of hepatitis-C-virus infection. Moreover the infection during pregnancy is often underestimated. METHODS: One hundred and seventy-eight anti HCV-positive pregnant women were investigated to evaluate the incidence of HCV infection and the possibility of sexual transmission of the disease to the partners. RESULTS: 126 patients out of 178 (70.8%) were positive for viral infection at PCR. In 96 patients (53.9%) HCV-positivity was detected for the first time in the actual pregnancy. 147 male partners out of 178 were checked for HCV-positivity and in 31 of them (21.1%) HCV antibodies were found. CONCLUSIONS: The results underline the importance of a screening for HCV-positivity in every pregnant, searching for anti-HCV antibodies also in patients not reporting risk factors. ALT values seem to be of little importance in the monitoring of the pathology. Sexual transmission of HCV virus from woman to man seems to occur rarely. PMID- 10379148 TI - [Abnormal uterine bleeding during climacteric. Correlation between transvaginal ultrasonography, hysteroscopy and histology]. AB - BACKGROUND AND AIMS: The authors evaluated the accuracy of ultrasonographic findings compared to hysteroscopic and histological results in the diagnosis of anomalous uterine bleeding in menopause. METHODS: Forty-eight women suffering from the above pathology and attending the Preventive Gynecology outpatient clinic of Department C of the Gynecology and Obstetrics faculty at Turin University during the period between September 1996 and July 1997, underwent first ultasonography using a transvaginal probe and then outpatient hysteroscopy with endometrial biopsy. RESULTS: A total overlap between the ultrasonographic image, hysteroscopic results and histological diagnosis was only obtained in the group of menopausal patients. CONCLUSIONS: In line with the data reported in the literature, the authors imposed a cut-off endometrial thickness of 4 mm above which further diagnostic tests were performed using hysteroscopy with targeted biopsy. PMID- 10379149 TI - [Intra-spermatic L-carnitine and survival of sperm motility]. AB - BACKGROUND: The aim of this work was to evaluate the intrasperm carnitine (L-C) content related to sperm motility survival in bovine cervical mucus and in culture medium (Tyrode solution). METHODS: The following subjects were selected: 15 fertile normospermic subjects (according to WHO guidelines) and 31 male partners of infertile couples (semen profile: volume > 2.0 ml; concentration > 20 x 10(6)/ml, progressive motility > or = 25% (WHO categories "a" and "b") after fluidification, abnormal forms < 70%; round cells < 1.0 x 10(6)/ml). After standard semen analysis, the samples were subdivided into three aliquots in order to carry out: a) intrasperm L-C (free, total and acetylated) assay; b) sperm motility survival in bovine cervical mucus; c) sperm motility survival in culture medium. RESULTS: A strict correlation was found between L-C (total and acetylated) content and motility survival in cervical mucus. This is probably due to the fact that in cervical mucus lipids are an important energy source for sperm and to metabolize these lipids intrasperm L-C is essential. Therefore, L-C content can be considered as an indicator of sperm motility life-span in cervical mucus. A significant correlation, even if reduced compared with cervical mucus, was also observed between L-C (total and acetylated) content and sperm motility survival in the biological medium. This probably is because the L-C system modulates the reserves of free CoA, essential to the tricarboxylic acid cycle function. CONCLUSIONS: The intrasperm L-C deficit could be due to; a) alterations in the L-C uptake mechanisms in the epididimys due to inflammatory processes; b) lack of testosterone (L-C uptake is androgen-dependent). Therefore, the therapeutic implication of this finding is that where hypomotility is due to intrasperm L-C deficit, exogenous L-C administration or improvement of L-C epididymal testosterone-dependent uptake could promote the acquisition of sperm motility. PMID- 10379150 TI - [The role of antenatal courses in preventing the onset of altered pelvic statics and urinary incontinence]. AB - BACKGROUND AND AIMS: This study was prompted by the frequent finding of urinary incontinence in peri- and postmenopausal women. In an attempt to reduce this incidence, the authors evaluated the role of antenatal courses as an important preventive and operative means of providing information aimed at preventing the onset of alterations to pelvic statics and urinary incontinence. The clinical data of 40 puerperae, of whom 45% had followed our antenatal course, were examined for this purpose. METHODS: A functional examination of the perineum was made in all subjects on discharge after birth and 6 months later. RESULTS: These findings appear to indicate that antenatal courses play an important role in preventing obstetric perineal complications. CONCLUSIONS: During the courses all women were given useful advice on hygiene and diet, including guidelines for suitable regular exercise, the need to drink about 1 litre of unsweetened liquid daily, a balanced diet and advice regardingf perineal relaxation. Subsequently, all subjects were advised to continue exercising after birth to ensure the correct rehabilitation of the perineal floor and they were also given a sheet containing the different learnt during the antenatal courses. In this way these subjects increased their awareness of the need not to underestimate the early symptoms of urinary incontinence should they occur after birth. PMID- 10379151 TI - [Preterm premature rupture of membranes in the 19th week of pregnancy. A clinical case]. AB - A case of premature rupture of membranes occurred at 19 weeks of pregnancy is reported. The patient underwent a genetic amniocentesis at 16 weeks of gestational age. During the hospitalization period, she presented amniotic fluid leakage responsible of olygohidramnios. She was treated with antibiotic, spasmolytic, tocolytic and cortisone therapy. All subsequent parameters were evaluated: blood cell count, heart rate frequency, body temperature. Ultrasound examinations were performed every two weeks. Cervical smears to detect infections were normal. Labour started at 28 weeks and the patient delivered spontaneously a 1010 g male baby, Apgar 4/8. The heaviest complication was a cerebral hemorrhage and the subsequent frontoparietal hematoma which progressively reduced. To date the neurological prognosis is good. PMID- 10379152 TI - [Cloacal exstrophy, ultrasonic diagnosis]. AB - This study aims to underline the importance of transvaginal ultrasonography in the first three months of pregnancy. The authors report a case which was referred to their attention for preliminary tests performed prior to amniocentesis, recommended by the general physician because of the mother's age. The primapara woman and father of the fetus were healthy and the family history excluded hereditary diseases or congenital pathologies. Transvaginal ultrasonography permitted the diagnosis at the start of the fourth month of pregnancy of a polymalformed fetus with cloacal exstrophy, sacral myelomeningocele, clubfoot, single umbilical artery. In this case, the mother decided to undergo eugenic abortion in view of the severity of the pathology. PMID- 10379153 TI - [A case of peritoneal tuberculosis. Contribution of mini-endoscopy]. AB - BACKGROUND: Tuberculosis in the genital and peritoneal region is increasing in last years. For this reason, the high value and efficacy of laparoscopy using a small caliber endoscope for the diagnosis of this disease, which often presents no specific and heterogeneous clinical features, is underlined. METHODS: A case of peritoneal tuberculosis in a young female presenting fever, asthenia and peritoneal effusion, is reported. Blood tests, X-ray and cytological examination of the peritoneal fluid all failed to point out the right diagnosis. Then multiple biopsies of the peritoneum and the external surface of uterus and ovaries were made using laparoscopy. RESULTS: Laparoscopy clearly showed the miliary nodules. The histology showed multiple granulomas composed by inner caseous necrosis and outer layer of epithelioid histiocytes and Langhans cells, leading thus to the diagnosis of peritoneal tuberculosis. The patient, treated with streptomycin and rifampicin, five months after diagnosis, did not show any feature of tubercular disease. CONCLUSIONS: Because of its safety, laparoscopy is a very useful and powerful diagnostic technique especially in those young women presenting with painful abdominal symptoms without any clear evident cause. PMID- 10379155 TI - [Mechanisms of progression of cerebral infarction]. AB - The progression of neurologic symptomatology in the first 48 hours after cerebral infarction appears in more than one third of the patients causing greater morbidity and mortality. Early detection of neurologic deterioration, identification of the predictive factors of worsening and knowledge of the pathogenic mechanisms responsible and their correction make up the basis of the most effective therapy in cerebral infarction. Different hemodynamic alterations (progressive occlusion and growth of the thrombus, insufficiency of colateral circulation, arterial hypotension, etc.) influence, although to different degrees in each patient, in all progressive infarctions. These mechanisms sequentially and progressively condition the necrosis of the areas of ischemic shadow and how new peripheral areas of healthy cerebral tissue are included in the ischemic region, progressively increasing the intensity or the extension of the neurologic manifestations. PMID- 10379154 TI - [Therapeutic posterior colpo-celiotomy. Authors' experience]. AB - BACKGROUND: To evaluate the advantages of posterior colpoceliotomy. METHODS: A retrospective analysis of the results of posterior colpoceliotomy performed between 1995 and 1998 for tubal sterilization, ectopic pregnancy or benign ovarian cyst. RESULTS: Surgery was possible in all 32 cases. The mean surgical time was 15 minutes. No intra- or postoperative complications were observed. The mean postoperative hospitalization lasted 32 hours. In 14 of 16 cases of ectopic pregnancy salpingectomy has been performed. In 2 cases tubal curettage has been performed with enucleation of contralateral dermoid cyst in 1 case. CONCLUSIONS: Colpoceliotomy is, in selected cases, a valid alternative to laparoscopy. PMID- 10379156 TI - [Neuroimaging in cerebral ischemia]. AB - A critical review about different neuroimaging techniques and its contribution to the field of cerebrovascular diseases is presented. The advances of neuroradiology have revolutionized stroke diagnosis. The inminence use of specific treatment for acute brain ischemia (neuroprotective and thrombolytic drugs) make neurologists become familiar with several imaging techniques that will also change stroke management. Nowadays, computed tomography is the mostly available technique for spanish neurologists in the acute phase, because of that, a suitable knowledge of early CT signs of acute infarction make possible use it in peak condition. Perfusion-and-diffusion-weighted magnetic resonance give such a lot of information for acute stroke, that at the near future, therapeutic decisions will be based on these findings. The role of other functional an angiography techniques is discussed. PMID- 10379157 TI - [Hypertensive vascular disease and cerebral microcirculation]. AB - Cerebral microcirculation has a series of complex relationships with arterial hypertension determined, on one hand, by the size and the location of the vessels involved, and on the other hand, by the chronic or acute nature of the hypertension. The small arterial vessels of the cerebral parenchyma react to the effects of chronic hypertension with irreversible structural changes, whose pathologic and radiological correlation is chronic ischemia of the white substance, shown by paleness of the white substance, together with small lacunar infarctions, with a clinical association of dementia, motor disorders and pseudo bulbar syndrome. With the appearance of an acute rise in arterial pressure, these vessels react with generally reversible changes which lead to an increase in the permeability of the hematoencephalic barrier with formation of cerebral edema and a clinical association generally demonstrating the focal nature of the vascular abnormality (such as in hypertensive encephalopathy with changes in posterior hemispheric predominance) or its unilateral location in cases in which the process occurs in one of the carotid territories (post-endarterectomy). PMID- 10379158 TI - [Carotid disease]. AB - Carotid atheromatosis is the most frequent disease of the internal extracranial carotid artery. Although important advances have been made in the knowledge of its pathophysiology, relevant points remain unknown. The need for early diagnosis is conditioned by the high sociosanitary load of secondary cerebral ictus. Modification and treatment of the vascular risk factors are necessary for both primary and secondary prevention. The current diagnosis is fundamentally based on Doppler ultrasonography and imaging studies in B-mode in real time, angio MR, helicoidal CT and overall on digital angiography. The need to treat the neurologically asymptomatic stenoses, greater than 70% is still questioned by the slight benefit provided. To the contrary, the need for surgical treatment by endarterectomy in stenoses of greater than 70% is well established. Careful analysis of the ECST and NASCET series, however, leads to the suspicion that at the time of making surgical decisions, a series of predictive variables of the long term results should be considered to individualize the cases which may most benefit from this treatment. On the other hand, given the surgical morbimortality it seems reasonable to search for therapeutic alternatives to endarterectomy. One of these is percutaneous transluminal angioplasty, the results of which appear to provide a valid alternative, specially in the cases which can not undergo surgery. PMID- 10379159 TI - [Heparin in acute cerebral ischemia]. AB - Non fractionated heparin is used in neurovascular disease because of its antithrombolytic action. Therefore, the design of the clinical studies performed to date has emphasized these actions. However, the results obtained in the most recent clinical studies have been discouraging, not only because they have demonstrated its lack of efficacy, but also because the use of this heparin has been followed by an excessive number of complications. This article provides critical comments on the possible causes of these results and underlines the most recent advances obtained in the knowledge of the mechanism of action of this drug. Particularly, its potential antiinflammatory role is emphasized as are the experimental requisites required for this antiinflammatory action to be fully expressed. Based on the results shown it is concluded that it is necessary to prospectively evaluate the role of heparin in acute ischemic ictus. PMID- 10379160 TI - [Thrombolysis in cerebral ischemia]. AB - Thrombolytic treatment with rt-PA is the first effective step in the therapeutic management of acute ischemic infarction. The benefit of rt-PA was clearly demonstrated in the NINDS study which showed a reduction in mortality or dependence in 13% of the cases. However, a reduction of only 3-5% as observed in the ECASS-II study is clinically relevant. In both the individual analysis and metaanalysis, the rt-PA studies indicate that despite a high frequency of symptomatic hemorrhages, mortality or morbidity do not increase and the clinical evolution improves. We believe that, at present, the question is not whether thrombolysis is effective in acute infarction, but whether we have well trained physicians in well equipped centers to provide this treatment adequately and safely. The patients admitted to stroke units within the first six hours should be considered for thrombolysis with rt-PA after careful evaluation of the inclusion and exclusion criteria. In Spain, however, rt-PA should be exclusively administered in accredited centers which have obtained authorization to perform its compassive use. In these cases, treatment with rt-PA should be carefully adjusted to the recommendations published by the AHA, it should be indicated by neurologists who are experts in the management of ictus, and administered in patients who may be closely monitored in stroke units. Hospitals without these possibilities should promote a radical change in stroke management. In these centers, neurologists, in collaboration with radiologists, internists and emergency physicians, should develop protocols of clinical performance, reorganize the priorities of the emergency departments, and familiarize themselves with CT findings and with the treatment criteria. The results of the metaanalysis of the finalized studies will allow better definition of the groups of patients at high risk of developing cerebral hemorrhage, and those in whom rt PA may be more effective. The potential risks and benefits of rt-PA should be individually evaluated and discussed with the patient and relatives before initiation of treatment. However, the existence of severe potential complications should not invalidate the benefit of thrombolytic treatment, similar to the fact that the severe surgical complications of carotid endarterectomy (5%) have not obscured the clear benefits of this intervention on the long term prognosis of the patients. PMID- 10379161 TI - [EEG monitoring of epilepsy]. AB - Electroencephalography (EEG) constitutes an integral part of the diagnostic process in epilepsy. It is the most important method of investigation in the management of epilepsy and has been widely developed in the last few decades. At present, technological development has provided us with the digital EEG and miniaturization of the equipment with which we are able to register as many channels as necessary in any circumstance and for an indefinite period of time, and together with the development of information technology we are now able to obtain rapid and effective analysis of large amounts of data and a reduction in the number of apparatus. These changes have revolutionized EEG. The EEG which has been used as a diagnostic procedure a posteriori, has increasing application in the direct monitoring of cerebral function. Prolonged EEG monitoring or that performed in the ambulatory, in intensive care units and the video EEG are increasingly accessible and necessary tools for the management of epilepsy. PMID- 10379162 TI - [Chandelier cells and their possible implication in epilepsy]. AB - Epilepsy is one of the most frequent human neurological diseases. Clinical symptoms of epilepsy are largely confined to the cerebral cortex but, given the scarce data available about the synaptic circuitry of the normal and epileptic human brain, the basic mechanism (or mechanisms) of human seizure activity is still a mystery. This disease can appear after multiple brain pathologies (e.g., tumors, vascular alterations, developmental abnormalities, etc.). However, these alterations are not intrinsically epileptogenic, since some patients displaying them are epileptic, whereas others are not. Thus, it is unknown how and why normal cortical circuits become epileptogenic. In this article, we explain the hypothesis that chandelier cells (which are considered to be the most powerful cortical inhibitory interneuron) may represent a key component in the etiology of epilepsy. PMID- 10379163 TI - [New epileptic syndromes]. AB - The recent and increasing interest for previously undescribed forms of epilepsy has been due to the description of families in which several affected members presented specific forms of epilepsy. Most epilepsies representing the description of new epilepsy phenotypes are partial epilepsies, although a new form of generalized epilepsy (generalized epilepsy with febrile seizures plus or GEFS+) has also been described. Our understanding of the clinical and genetic characteristics of the new familial epilepsy syndromes and the identification of informative families should accelerate the discovery of the basic mechanisms implicated in the production of partial seizures. The recent description of the syndrome of autosomal dominant nocturnal frontal lobe epilepsy, its localization to chromosome 20, the identification of the responsible gene (the alpha 4 subunit of the nicotinic cholinergic receptor) and the characterization of a mutation in two families are a good example. The recognition of the new epilepsy syndromes is of great interest for clinical neurologists and should lead to the establishment of more precise prognoses and therapies. In those families with several affected members, genetic knowledge may be important for genetic counseling purposes. PMID- 10379164 TI - [Rational choice of antiepileptic treatment]. AB - The choice of the adequate antiepileptic treatment is based on the clinical experience more than rationality. During some decades, the combination of two antiepileptic drugs was considered the initial treatment but monotherapy showed more advantages (effectiveness, fewer adverse events, fewer teratogenic effects and better compliance). New antiepileptic drugs have increased our interest and knowledge of the epilepsies. They have changed some of our therapeutical schemes. Sodium valproate continues to be considered the choice treatment for all the idiopathic, cryptogenic and symptomatic generalized epilepsies. Lamotrigine and topiramate are two valid alternatives in these epileptic syndromes. In West's syndrome vigabatrin is considered the initial treatment. Carbamacepine, vigabatrine and tiagabine are not indicated in the treatment of generalized idiopathic epilepsies especially in patients with absence seizures. In focal epilepsies, both cryptogenic and symptomatic all the antiepileptic drugs have shown efficacy and the choice treatment is based on the adverse events and the teratogenic power. Prospective studies in patients with the same type of seizures and epileptic syndromes will allow us to determine the more adequate antiepileptic treatment. PMID- 10379165 TI - Epilepsy and malformations of the cerebral cortex. AB - Abnormal cortical development is increasingly recognized as a cause of human epilepsy. Development of the cerebral cortex involves three distinct but overlapping processes consisting of neuronal and later glial proliferation, neuronal migration and cortical organization. Cortical malformations can originate from abnormalities of any or all of these processes. Certain malformations are known to be genetically determined, while for others a genetic origin has been hypothesized. In addition, there are some that may be linked to prenatal insult. Some malformations are notably more epileptogenic than others. In specific forms, epileptogenesis appears to originate from intrinsic properties of the dysplastic tissue, and is stereotyped in expression. Cortical malformations are also known that are affected by age-related influence and may be accompanied by different epileptic syndromes, sometimes with good long-term outcome. In patients with intractable seizures which are symptomatic of a cortical malformations it is necessary to diagnose the type of malformation and the associated epileptic syndromes and to assess the characteristics of epileptogenesis. Planning of surgical treatment of the associated epilepsy must rely on careful evaluation of all these informations. PMID- 10379166 TI - [Training program for rehabilitation of patients with internal diseases]. AB - Physical training for patients with internal diseases differs in many points to the physical activity recommended for health maintenance. Cardiac patients are usually limited by symptoms (angina, ECG abnormalities, anaerobic threshold) therefore the intensity of the training (monitored by heart rate or ECG) must be setted by an ergometer exercise testing. Patients with obliterative peripheral artery disease may surpass the local anaerobic threshold during interval-type loads. Blood pressure limits the training intensity of the hypertensive patients if not an organ lesion. COPD patients use the rest periods of an interval training for expectoration and for restitution of their blood gas values. In insulin dependent diabetes the vascular complications can be avoided by a proper insulin regime, training and diet. Day-to-day training by an even energy need acts like the insulin therefore it must be carefully dosed. In NIDDM also the carbohydrate metabolism can improve significantly. In anxiety and depression the training and the social milieu offers a physiological trigger for the improvement. Other rehabilitative interactions (psychology, dietetics, behavioral modalities etc.) are built up in the basis of exercise training. PMID- 10379167 TI - [Ganglioneuroma of the adrenal gland]. AB - 149 patients with adrenal incidentalomas were examined. Sixty-eight cases were histologically confirmed, five of them had ganglioneuromas. On the basis of these patients history current knowledge of this benign tumour was summarized. Histological and pathological characteristics of one tumour suggest that ganglioneuromas may develop by maturing of malignant neuroblastic tumours. The clinical symptoms (abdominal pain, meteorism) were local. In 2 of 5 cases mildly elevated levels of urinary vanillylmandelic acid and catecholamine could be measured. One patient had persisting hypertension after surgery. In an other patient previous diarrhoea stopped after the removal of tumour. On the basis of ultrasound and computertomographic features, the size and origin of a tumour and its relation to the surrounding organs can be well characterized. One patient was inoperable because of an infiltratively spreading tumour, but during five years of follow-up no tumour progression could be observed with computertomography. After surgery we could follow only 2 of 4 patients. Until now no recurrence of tumour were detected. PMID- 10379168 TI - [Quality control of medical documentation of in-patients at the St. Stephen Municipal Hospital in Budapest]. AB - Within the frame of hospital quality control programme, the authors have investigated since 1997 the medical documentation concerning in-patients of St. Stephen Hospital Budapest. An assessment form of 26 items related to main elements of medical documentation has been created for the study. Choosing of both order of departments' succession and cases to be investigated took place at random. Evaluation was performed at a scala of 0-3 points, by a five-membered work-group; all members first carried out the qualifications independently of each other, and then they formed the final opinion together at common sessions. Investigation of 204 "first round" cases proved the feasibility of the applied method. As it turned out from the results, the quality of documentation showed statistically significant differences between departments of the hospital and the average standard of documentation could not be declared as satisfying at the period of basic survey. Deficiencies of documentation especially in respect of case history, disease course, personal data and informed consent have been found. On the other hand, registration of status on admission and final report got a relatively high average score-number. The authors call attention to the importance of ordinary control of medical documentation. PMID- 10379169 TI - [Determination of serum S-100 protein and 5-S-cysteinyl-DOPA levels in melanoma patients]. AB - This was the first time that authors detected se-S-100 and 5-SCD values with patients with malignant melanoma in Hungary. They examined the change of serum S 100 and 5-SCD value parallel. Sera were obtained with 184 melanoma patients 326 times. Patients were ranked into groups on the basis of clinical symptoms: free of symptoms and suffering from it (primary tumour, regional lymph node metastasis, soliter or multiplex distant metastasis). On the basis of the initial results the following have been found: S-100 protein and 5-SCD serum levels had no prognostic value in patients with primary melanoma. Patients without symptoms showed values around the normal level. There was significant difference in both markers between patients with or without symptoms. Significant differences were found between clinical stage I and II, as well as in clinical stage II and III. In the case of S-100 protein there was significant difference between the values of patients with soliter and multiplex distant metastasis. PMID- 10379171 TI - [Drug-addicted students: instead of jail, delay of driver's licence]. PMID- 10379170 TI - [Benign tumor of the small intestine causing recurrent hemorrhage]. AB - The authors reported a 56-year-old man, who suffered from repeated massive gastrointestinal bleeding. The source of the bleeding was leiomyoma of the ileum. Abdominal computertomography and selective enterography confirmed the tumour's localisation. After partial resection of the ileum the patient had no more complaints. PMID- 10379172 TI - [Questions about the costs of therapy of reflux esophagitis]. PMID- 10379173 TI - [DNA-endonucleolysis in cell nuclei of brain and liver in patients dying from hemorrhage and after resuscitation: general patterns and differences]. AB - Specific features of Ca2+, Mg2+ dependent DNA endonucleolysis in the nuclei of the cerebral cortex, hypothalamus and liver were investigated in mongrel anesthetized male and female dogs. The endonucleolysis was studied in different periods of long-term arterial hypotension and in postresuscitation period, with strain pUC 19 plasmids as substrate for determination of nuclear endonuclease activity. It was established that nuclear DNA-endonucleases coupled with chromatin activated earlier in brain cortical and hepatic neurons than in the hypothalamus. Changes in activity of the enzymes directly correlated with duration of CNS ischemia. Active endonucleolysis occurred in cerebral and hepatic nuclei even 3 months after the blood loss and resuscitation. Postresuscitation changes in Ca2+ and Mg2+ dependent endonucleases in cortical nuclei are phasic while in the liver their activity for three months did not differ much from that in the end of hypotension. The activity of nuclear endonucleases in the hypothalamus returned to normal after beginning of resuscitation and did not change later. The data obtained evidence for active involvement of apoptosis mechanisms in brain and liver cell degeneration in massive blood loss and in postresuscitation period including a late one. PMID- 10379174 TI - [Action of neuropeptides on the status of neuronal populations in the post resuscitation period: structural-functional correlation]. AB - Effects of some neuropeptides and hormones (oxytocin, melanostatin, oxytocin in combination with estradiol, somatostatin) on neurons of the V cortical layer, Purkinje cells of medial and lateral regions of rat cerebellum were studied in rats after 15 minutes of cardiac arrest. A single administration of the peptides after successful cardiopulmonary resuscitation improved the condition of neuronal populations (prevented dystrophy and cell death), accelerated neurological recovery. One of the mechanisms of action of oxytocin, melanostatin and oxytocin in combination with estradiol is elevated number of satellite glial elements of the nervous tissue. Responses of varying neuronal populations to neuropeptide action are different. These data are essential for design of pathogenetic methods of prevention and treatment of posthypoxic encephalopathies. PMID- 10379175 TI - [Post-resuscitation neurosomatic deviations and the feasibility of their correction with a drug of metabolic action]. AB - Circulation in white rat males was stopped for 10 minutes by ligation of the intrathoracic vascular bundle of the heart. Proxipin in a dose 10 or 20 mg/kg was injected i.p. after restoration of effective cardiac activity and the next 3 days. Proxipin improved resuscitation outcomes, relieved symptoms of CNS excitability 4-6 months after resuscitation, changed adaptation to stress agents, protected against risk-factors of atherogenesis. However, proxipin-induced inhibition of the motor activity suppressed the free-choice ability to find food. PMID- 10379176 TI - [Role of disorders of metabolism in rat myocardium and endotoxemia in the pathogenesis of post-resuscitation cardiodepression]. AB - The model of 4-min clinical death due to acute blood loss was used to study cardiac contractility, myocardial metabolism and causes of endotoxemia in early postresuscitation period. The investigations were made on the whole body, isolated, isovolemically contracting heart and isolated papillary muscle. A marked reduction in functional myocardial reserves, maximal within the first 24 hours of postresuscitation, with dominant defects in relaxation was seen. Pathogenetic factors responsible for cardiodepression are the following: hypoxia, impairment of bioenergetics, hyperactivation of lipid peroxidation, acidosis, membrane destruction, endotoxemia. PMID- 10379177 TI - [Application of the principles of the systemic approach to interpretation of the pathogenesis of traumatic shock and traumatic disease]. AB - Experimental and clinical studies for many years led the authors to the conclusion that systemic approach seems most adequate for current interpretation of pathogenesis of traumatic disease (TD) and its component--traumatic shock (TS). It seems valid to assess TD and TS severity and dynamics by the proportion of pathological and adaptive reactions. Drugs should be used for maintenance and stimulation of adaptive reactions. Strong correlations of dynamics and severity of TD and TS enable prediction of TD course and outcome in acute period by time criterion (+)-T and distinguish three degrees of TD severity as well as evaluation of probability of favorable or unfavorable (lethal) outcome. PMID- 10379178 TI - [Effects of presensitization and clinical death in the history on the dynamics of morphofunctional parallels of retinal and cerebral circulation]. AB - A negative effect of preliminary sensibilization with a normal serum and clinical death (restoring to life according to V. A. Negovsky et al.) on cerebral blood supply and eye retina within 5 weeks of postresuscitation period was studied on dogs. Microcirculation disorders in the groups of sensibilized dogs were more prominent as compared to intact animals. The diameter changes of pial and retinal microvessels did not correlate, but qualitative alterations in retinal and cerebral microvessels were of the same type. PMID- 10379180 TI - [Hemocoagulation while dying of blood loss and restoration to life after clinical death]. AB - Hemostatic system function was studied in dogs dying of acute blood loss and restoring to life after 4-min clinical death. Phasic changes in hemostatic system of two and three types occurred in the blood loss and reanimation, respectively. Dogs with favorable postresuscitation period exhibited hypercoagulation when dying, hypocoagulation 1 hour after reanimation and normal coagulation 3-6 hours after clinical death. PMID- 10379179 TI - [Transthoracic defibrillation of cardiac ventricles: effects of novocainamide on effectiveness of an electric impulse of bipolar sinusoidal shape]. AB - Experiments on nembutal-narcotized dogs were made to study dose effects of novocainamide (10-65 mg/kg) on threshold values of transthoracic defibrillation current of bipolar sinusoidal shape. Reversible dose-dependent increase of the threshold of ventricular defibrillation by 21%, on the average, was achieved only after novocainamide lowering of arterial pressure by 15-35 mm Hg. Coefficient of correlation between the drug dose and intensity of the efficient current made up 0.72. PMID- 10379181 TI - [Segmental contractile activity of the left ventricle in life threatening arrhythmia]. AB - Intramural pressure (IMP) in the middle segment of the left ventricle and in the apex under ventricular extrasystoles was measured in experiments on the hearts of narcotized dogs. Early extrasystoles caused greater lowering of IMP and blood ejection. Bigeminia was associated with segmental and transmural nonuniformities of IMP which may be due to additional dragging effects in the myocardium. Magnitude of cardiac ejection under extrasystoles depends on nonuniform contractions of the segments of the left ventricle. PMID- 10379182 TI - [Functional-morphologic evaluation of the effect of the regulatory peptide kyotorphin on the status of the CNS in the post-resuscitation period]. AB - Functional and morphological analysis of changes in white rat CNS was made throughout 4 months after 12 minute interruption of circulation. Within 3 months after clinical death and resuscitation the rats' neurological status was normal except some abnormalities in emotions and motives especially noticeable under stress. Moreover, neurons in the hippocampus and cerebellum were destroyed. A single intranasal 0.05 mg/kg dose of kyotorphin--a peptide isolated from the brain of winter-sleeping animals--given 30 min after beginning of resuscitation increased survival, accelerated restoration of neurological status, normalized emotional reactivity, orientation and search behavior, prevented death of neurons in the hippocamp and cerebellum. PMID- 10379183 TI - [Organization and education for cardiopulmonary resuscitation]. PMID- 10379184 TI - [Learning cardiopulmonary resuscitation using conventional external cardiac massage or active compression-decompression in simulated cardiopulmonary resuscitation]. AB - OBJECTIVE: To compare medical students' simulated learning of two different techniques of cardiopulmonary resuscitation (CPR). One was conventional external cardiac massage (ECM) and the other was active compression-decompression (ACD CPR). MATERIAL AND METHODS: The study group (group S) comprised 111 students enrolled in their fourth year of medical studies who had no prior experience of CPR. Group R, the control group, was made up of 32 medical residents in anesthesiology and post-anesthetic intensive care. Before the study, group S received 5 hours of theoretical classes on CPR and both groups saw a video explaining each technique just before performing the test. All subjects applied each method to an adult dummy for one minute. The variables evaluated were frequency of complete and effective thoracic compressions and the body weight of the resuscitator. RESULTS: Each group performed similarly using the two techniques in terms of frequency of total compressions achieved. For each technique, the number of effective compressions achieved by group S (49.4 +/- 22.9 with ECM and 42.5 +/- 20.7 with ACD-CPR) was significantly lower (p < 0.05) than the number attained by group R (71.2 +/- 18.6 with ECM and 58.8 +/- 12 with ACD-CPR). Group R's frequency of effective compressions was significantly higher (p < 0.05) with CPR than with ACD-CPR. Body weight had no influence on the number of total compressions or efficacy in group R, whereas lower body weight in group S was significantly related to lower frequency of effective compressions with ECM p < 0.05). Neither group achieved a frequency of 80 total compressions in one minute. CONCLUSIONS: With the present teaching method, the medical students' performance was poor for both types of CPR and was affected by body weight. The residents' performance was less effective with ACD-CPR, a technique that was new to them, than with conventional ECM, with which they were expert and on which body weight had no impact. PMID- 10379185 TI - [Psychological distress and preoperative fear in surgical patients]. AB - OBJECTIVE: To study the prevalence of psychological disorder, cognitive deterioration and anxiety in patients undergoing surgical procedures with general anesthesia. PATIENTS AND METHODS: A representative sample (n = 450) of surgical patients at a tertiary hospital was selected, excluding patients with a history of mental illness or drug use, and those with cancer. After admission, the day before surgery, we collected demographic, medical and surgical data and administered the Spanish versions of Folstein's Mini Cognitive Examination (MCE) and Goldberg's General Health Questionnaire (GHQ). The patients were also asked if they felt anxiety about the surgical procedure and what they feared. RESULTS: The prevalence of cognitive deterioration (MCE) was 8.7% and the prevalence of psychological disorder (GHQ 28) was 29.8% (higher for women). Combining the two instruments, 38.5% showed relevant psychological disorder. Some type of anxiety was expressed by 60.9%, with the fear of "not waking up" being the most common (26%). CONCLUSIONS: The prevalence of psychological disorder is somewhat lower than that reported by other authors for presurgical patients, probably because our study enrolled patients with no history of mental illness related to other causes. The prevalence of anxiety found is similar to that reported in the literature. PMID- 10379186 TI - [Evaluation of amplified spontaneous pattern ventilation in postoperative patients. Comparison with pressure support]. AB - HYPOTHESIS: Amplified spontaneous pattern (ASP) ventilation is a new method for giving partial support by reproducing, in an amplified manner, the patients' own spontaneous flow wave form, thereby optimizing patient adaptation to support. OBJECTIVES: To study clinical use of ASP ventilation for the first time in terms of flow wave form, patient synchronization, ventilation pattern, work of breathing (WOB), and inspiratory effort by transpulmonary pressure (TPP) and to compare ASP and pressure support ventilation applied in a similar clinical setting. PATIENTS AND METHOD: We studied 20 patients after heart surgery during weaning from controlled ventilation. Each patient was ventilated during 4 phases of 15 min each with two similar levels of support using ASP and PS applied successively and randomly. Maximum support (ASPmax and PSmax) was that which was set to give the same respiratory frequency (F) and tidal volume (VT) as that recorded during the earlier period of controlled ventilation. Half support (PEA1/2 and PS1/2) was set for half the aforementioned levels. At the end of each phase we obtained gas measurements and flow (V) curves and VT and pressure in airways and esophagus (Pes) to measure F, VT, the ratio of inspiratory to total time (TI/TTOT and TPP, as well as the VT/Pes loop with a mechanical ventilation monitor. The WOB was determined by measuring area under the curve (Campbell's method). RESULTS: We observed no significant differences between the two modes, with similar levels of support, with regard to ventilation (PaCO2) or ventilatory pattern (F, VT, TI/TTOT). De-adaptation occurred, however, eight times with PS (25%) but never with ASP. WOB and TPP decreased with PS when level of support increased, whereas with ASP these variables were constant regardless of level of amplification within the normal range. CONCLUSIONS: Adaptation to support is better with ASP than with PS during postoperative weaning and causes no significant respiratory work overload. PMID- 10379187 TI - [Hospital organization of cardiopulmonary resuscitation]. AB - That hospital cardiopulmonary resuscitation (CPR) should be supported by an organized plan rather than on the skills of individual health care personnel is a universally agreed-upon principle. Such a plan should guarantee that needed materials are available and in working order in all departments and that the team assigned to carry out CPR arrives promptly. Personnel other than the specialized team should also receive CPR training appropriate to their posts. The main features of a CPR plan are related to the five steps in the chain of survival: a) identification of a patient to be resuscitated, a matter that has important ethical ramifications; b) early recognition of cardiac arrest; c) early defibrillation; d) basic CPR, and e) advanced CPR. The CPR plan should incorporate the automatic recording of system, population, event and outcome variables. Task forces responsible for establishing and maintaining the plan and its quality control will periodically review the data with the aim of detecting errors, correcting them or introducing improvements. Various international societies and CPR committees have recently suggested a uniform way (the Utstein style) of recording and presenting data to allow comparisons either from hospital to hospital or over time within a single center. PMID- 10379188 TI - [Anesthesia and intraoperative treatment in 2 cases of simultaneous liver, pancreas, and kidney transplantation]. AB - We report two cases of patients who underwent simultaneous triple transplants (liver-pancreas-kidney) using organs taken from a single donor in each case. The anesthetic technique and perioperative treatment of each patient is described. The favorable evolution in both cases seems to indicate that although this type of transplant may be more complex, it is nevertheless a good therapeutic option for patients suffering terminal liver failure, kidney failure or diabetes type I. PMID- 10379189 TI - [Octreotide and a serotonin-secreting glomus tumor]. AB - Glomus tumors arise in the chemical receptors of vessels in the tympanic and jugular regions. Clinical signs depend on location, the structures invaded and a tumor's ability to secrete active amines and peptides. A 44-year-old woman was scheduled for excision of a serotonin-secreting tympanic glomus tumor. Urinary excretion of 5-hydroxyindolacetic acid (5-HIA) in urine over the previous 24 hours was 80 mg (normal < 10 mg). The patient received oral diazepam, ranitidine, intravenous diphenhydramine and subcutaneous octreotide (150 micrograms). Anesthesia was induced with propofol, alfentanil and vecuronium. The tumor produced an episode of bronchospasm and cutaneous rubor during surgical manipulation of the tumor. Airway pressure increased to 42 cmH2O and SpO2 decreased to 89%. Hypotension and bradycardia appeared. Once it was suspected that the symptoms stemmed from tumoral secretion of active substances, 20 micrograms of intravenous octreotide was administered. The bronchospasm decreased and hemodynamic changes were resolved in three minutes, with no recurrence of symptoms. The patient received 100 micrograms of octreotide subcutaneously every 8 hours throughout the 72 postoperative hours. Urinary excretion of 5-HIA was 12 mg on the fifth day and the patient was released without having experienced complications. Appropriate preoperative preparation is important in patients with such tumors, as are early detection of respiratory and hemodynamic changes that may occur during surgery and correct perioperative treatment. Octreotide, a longer-lasting somatostatin analogue, has facilitated the handling of such cases. PMID- 10379190 TI - [Effectiveness of adenosine as a pulmonary vasodilator during weaning from extracorporeal circulation after mitral valve replacement in a patient with severe pulmonary hypertension]. AB - A 75-year-old woman with mitral stenosis and tricuspid insufficiency underwent mitral valve replacement surgery. After insertion of a catheter into the pulmonary artery, upper respiratory system pressure was seen to rise above systemic arterial pressure. To relax the pulmonary vessel, we administered adenosine, a drug that is metabolized rapidly during its first pass through the lungs before hypotension or other adverse side effects occur systemically. Adenosine decreased pulmonary artery pressure and vascular resistance, optimizing right cardiac function with successful output to extracorporeal circulation. PMID- 10379191 TI - [Evaluation of transfusion in pulmonary surgery]. PMID- 10379192 TI - [Intubation problems--ventilation problems]. PMID- 10379193 TI - [Queen Victoria's anesthesias]. PMID- 10379194 TI - [Contributions of Carlos Roe Leon to obstetric anesthesia and analgesia in Spain at the beginning of the 20th century]. PMID- 10379195 TI - [The placebo effect: classes of explanation]. AB - The placebo effect is a frequent phenomenon in medicine, but very little is known about its mechanisms. An overview is given of the different classes of explanation of the placebo effect in analgesia and in particular the role of endogenous opioids, classical conditioning and expectations. Then the question is raised which are the properties of placebo for which a theory has to provide answers in order to be coherent. These properties are, between others, the efficacy of placebo in a variety of conditions, in individuals with different personality characteristics, etc. Finally, the difficulty of observing individual placebo is emphasized and problems concerning the diagnostic and therapeutic use of placebo are mentioned. PMID- 10379196 TI - Oxidized low-density lipoprotein enhances intimal thickening and alters vascular reactivity. AB - Oxidized low density lipoprotein (oxLDL) is present in atherosclerotic lesions and has been implicated in the etiopathogenesis of atherosclerosis mainly based on in vitro studies. In view of the lack of data on the activity of oxLDL in vivo, we decided to study its effects in the rabbit by local application at the level of the vascular wall. Intimal thickening was evoked by the placement of a silicone collar around the carotid arteries during 2 weeks. The collar was connected to an osmotic minipump containing human oxLDL (7 micrograms h-1), LDL (7 micrograms h-1) or phosphate-buffered saline. Collar placement resulted in a thickening of the intima thereby increasing the thickness from 5 +/- 1 to 26 +/- 5 microns with the appearance of alpha-actine positive smooth muscle cells. Perivascular infusion of LDL or oxLDL significantly enhanced the intima, containing large amounts of T-lymphocytes, collagen and smooth muscle cells. The placement of the collar and the infusion of oxLDL during 14 days resulted in an increased sensitivity to serotonin and a decreased sensitivity to acetylcholine. The maximal relaxation to acetylcholine was reduced by 50% whereas the endothelium-independent relaxation to nitroglycerin were not affected. These results show for the first time that the local application of oxLDL in vivo promotes intimal thickening and impairs the endothelium-dependent relaxations thereby supporting the suggestion that oxLDL plays an important role in the morphological and functional changes present in atherosclerotic blood vessels. PMID- 10379197 TI - [Growing divergence inside primate T cell lymphotropic viruses]. AB - During our studies on the evolution of the PTLVs, we have isolated and genomically characterized two divergent simian T-lymphotropic viruses, not belonging to the previously well-established PTLV lineages. STLV-PH969 has been isolated from an Eritrean sacred baboon (Papio hamadryas), with an HTLV-like indeterminate serological profile. The entire 8916 nt long genomic sequence, was obtained from a cDNA library of PH969 mRNA, in combination with a PCR based strategy. All open reading frames (ORF) common to all HTLV related viruses could be identified without important deletions or insertions. Sequence comparison of the STLV-PH969 genomic sequence with the HTLV-I and -II prototype sequences revealed that this virus is equidistantly related to HTLV-I and -II. A phylogenetic analysis on the envelope and regulatory Tax proteins conclusively proved the ancient separation of HTLV-I, -II and STLV-PH969. Together these data provided enough evidence to justify the classification of this virus as a new type of primate T-lymphotropic virus, designated PTLV-L. We isolated a second highly divergent virus from pygmy chimpanzees (bonobo's, Pan paniscus) housed in the Antwerp Zoo. The animals infected with this virus showed an aberrant HTLV-I like serological profile. The entire 8855 nt long genomic sequence of the STLV PP1664 provirus was obtained by sequencing of overlapping PCR fragments. On this sequence, all ORFs common to all HTLV related viruses could be identified without any important deletions or insertions. Sequence comparison showed that the STLV PP1664 proviral sequence was related to HTLV-II and the virus is called STLV-II. However, in all genomic regions, STLV-IIPP1664 was much more divergent from any of the HTLV-II subtypes than these are from each other. In order to further investigate the genomic relationship of STLV-LPH969 and STLV-IIPP1664 with HTLV-I and -II, we also analyzed the genomic organization of the pX region, which differs between HTLV-I and HTLV-II. In the STLV-LPH969 and STLV-IIPP1664 producing cell lines, 2 and 5 viral messengers could be detected respectively, potentially expressing 3 and 5 different accessory proteins respectively. The splicing pattern and the resulting proteins differ from those of HTLV-I and HTLV II. In the light of the growing interest for xenotransplantation, the prevalence of distinct PTLVs in the wild and their capacity to cross the species barrier deserves further attention. PMID- 10379198 TI - [Lipid degradation by way of beta and alpha oxidation in peroxisomes of mammals]. AB - Recently, it has become clear that the peroxisomal beta-oxidation system in rat and man consists of multiple pathways. In rat and man straight chain fatty acids, dicarboxylic fatty acids and prostaglandins are oxidized via the L-specific pathway catalyzed by palmitoyl-CoA oxidase, multifunctional protein-1 and thiolase. 2-Methyl-branched fatty acids and the bile acid intermediates are oxidized via the D-specific pathway. In the rat this pathway is catalyzed by prostanoyl-CoA oxidase, multifunctional protein-2 and sterol carrier protein-X (branched fatty acids) and by trihydroxycoprostanoyl-CoA oxidase, multifunctional protein-2 and sterol carrier protein-X (bile acid intermediates). In the human, branched fatty acids and bile acid intermediates are oxidized via branched chain acyl-CoA oxidase, multifunctional protein-2 and sterol carrier protein-X. All enzymes of these pathways have been purified, cloned and characterized. Also the reactions that constitute the alpha-oxidation pathway for 3-methyl-branched fatty acids, have recently been identified in rat and man. The revised pathway consists of the following reactions: 1) an activation reaction catalyzed by an acyl-CoA synthetase, that forms a 3-methylacyl-CoA; 2) a hydroxylation (dioxygenase) reaction catalyzed by a 3-methylacyl-CoA 2-hydroxylase, that converts the CoA ester to a 2-hydroxy-3-methylacyl-CoA; 3) a cleavage reaction catalyzed by a 2 hydroxy-3-methylacyl-CoA lyase, that releases a 2-methyl fatty aldehyde and formyl-CoA. The branched aldehyde is dehydrogenated by an aldehyde dehydrogenase to a 2-methyl branched fatty acid that can be degraded by peroxisomal beta oxidation. Formyl-CoA is enzymatically hydrolyzed to formate which is then converted to CO2. PMID- 10379199 TI - [Yersinia pestis. Bacteriology]. AB - The author discusses the evolution in the classification of the bacterium, responsible for plague: first a classification based on phenotypic characteristics, later based on genotypic characteristics, to finally arrive at an evolutionist classification. He treats the seven species of the genus Yersinia that can be distinguished by DNA hybridization. He examines the issue of sequencing and decoding the chromosome and mentions research regarding the phenomenon that the metabolism of the organisms modifies as a reaction to signals of their changing environment. Furthermore the author discusses the efforts to characterize the strains of Y. pestis (antiqua, medievalis and orientalis). Finally he comments on the discovery of a multiresistant strain, isolated in 1995 in Madagascar. PMID- 10379200 TI - [Plague in southern Netherlands during the Middle Ages and modern times. Complaint about the status of the disease in its socioeconomic context]. AB - In this contribution the latest insights with regard to the demographic impact of plague in the Netherlands are discussed, although is remains difficult to clearly distinguish this factor from other causes of mortality. When, how and why did the plague reappear in Europe after several centuries of absence to become endemic for the next three centuries? When and why did it disappear in Western Europe in the seventeenth century? The first epidemics of plague probably were not as catastrophic in the Netherlands as they were in many other parts of Europe, which is remarkable since the Netherlands was, together with Northern Italy, the most populous region of Europe. The explanation is to be found in the general socioeconomic context, that was in many regards better than in the neighbouring regions. The 'crisis of the late Middle Ages' was not as deep in the Netherlands as elsewhere: a relationship demography--economy is therefore probable. Nevertheless, mortality was high, partly because of plague, but also because of other diseases--for which the common term pestilentia was in use. Some unique statistical data for Flanders illustrate this mortality from the late Middle Ages onward. The succession of mortality leading to a high average mortality rate was more important than accidental mortality, that could have a spectacular but often not long-lasting impact. PMID- 10379201 TI - [The persecution of Jews during the time of the plague (1349-50) in southern Netherlands]. AB - Research on the persecutions of the Jews at the time of the 'Black Death' in the Southern Low Countries has overemphasized the responsibility of the flagellant movement. In fact, the specific role of the flagellants was confined to spreading the rumor that the Jews were conspiring to kill all Christians by well-poisoning. A closer look at three examples--the persecutions in Hainaut, Brussels and Leuven -reveals that the driving forces behind the pogroms and executions were local and municipal authorities, while the countess of Hainaut and the duke of Brabant were too weak politically to defend the lives of their Jewish subjects. PMID- 10379202 TI - [Eye witnesses and the flagellants in the year 1349]. AB - Deeply affected and often desperately afraid, many contemporaries recorded their observations and emotions. These reports--no matter how obviously subjective they sometimes were--provide valuable information about what happened during the plague pandemic of 1348-1350. Thus many of our fellow countrymen left behind a direct testimony: Bartholomew of Bruges, a canon in Andenne; Gilles li Muisis, the abbot of Saint Martin in Tournai; Ludovicus Sanctus of Beringen; Simon de Couvin, a canon in Liege; Jan van Boendale, an alderman's clerk in Antwerp; John of Burgundy (also known as John of Mandeville), professor of medicine in Liege; but also texts in Middle Dutch that were not known up to now, and therefore not published, such as the important thesis by Arent Schryver, licentiate in medicine (see next article); an account in verse in the Brabant Chronicle, as well as contemporary testimonies in a different language that have been translated into our language, such as that by John of Eschinden, Johannes de Rupescissa or Guy de Chauliac (who had had the plague himself). They describe the precautions, the causes (God, a comet, an eclipse of the sun, the polluted water, the planets, the air), the symptoms, the social groups most likely to be affected (the youth, the lower classes, the clergy), the high mortality, the problems of hygiene,the social and administrative chaos, the general panic, the flight of countless people. One of the most virulent reactions led to the emergence of the flagellant sect. They originated from Hungary and advanced in an unstoppable advance with a growing number of followers as far as our country, singing, praying, dancing and flaying themselves until they drew blood. We only recently discovered what they sang in Dutch: very recently, a unique roll of parchment was discovered that they carried in their processions, and that contains the text of their songs and a flagellant sermon. The existence of this valuable document and its contents are presented here for the first time. PMID- 10379203 TI - [The plague regimen of Arent Schryver]. AB - This article is concerned with the edition of a 15th century plague treatise in Middle Dutch, by Arent Schryver, licentiate in medicine, from Dalen (Drenthe, the Netherlands), that has been discovered recently. The text has been found in manuscript number 33 of the Furstliche Salm-Salmsche Bibliothek in Anholt (FRG). It is the most detailed plague treatise in Middle Dutch known to date. Its structure and contents are clearly based upon the Compendium de epidimia of the Medical Faculty of Paris, but on numerous occasions it diverges from its model. Therefore it deserves its own place among the numerous plague treatises. PMID- 10379204 TI - [All around the deathbed of Lubbert ten Busch. The Modern Devotion and the plague in Deventer in 1398]. AB - The seriousness of plague epidemics can be expressed in numbers and medical terms, but we get closer to the past, and it becomes our own history more than in any other way, when we can empathise with the story of a single individual. In the summer months of 1398, the plague raged in the city of Deventer, which lies on the river IJssel, in the east of the present-day Netherlands. This plague epidemic also threatened the small community of priests and minor clerics which gave rise to the new spiritual movement of the Modern Devotion. This new community was concerned about its survival. Their vocation required that the brethren should help the citizens who remained behind. However, this could prove to be their undoing, and therefore it would be better to leave the city. They resolved the dilemma by dividing into two groups. Half of the brethren left to ensure the continuity of the community; the other half stayed in Deventer to help the people. The two groups stayed in contact by means of letters, the text of which has survived in several sources. The fears and forebodings became reality. The plague also affected the new community of brethren. The deputy rector, Lubbert ten Busch, also died of the plague. There is a letter from him which he wrote just before he died. This farewell letter was sent together with a letter from one of the brethren, saying that Lubbert had meanwhile died, and describing the scene of his death. Because of the personal tone and the tragic of content, the letters about the death of Lubbert ten Busch are unique medieval documents. They give a good insight into the way in which the plague could personally affect someone and his immediate companions in the late Middle Ages. This paper focuses attention on these letters, and reconstructs the events around this death, so that the letters speak to us once again. PMID- 10379205 TI - [Some bibliographies of pestilences. Johannes Jacobi in Antwerp (circa 1484-circa 1491)]. AB - One of the many works on the plague is by the Frenchman Johannes Jacobi. It dates from the 14th century and has been passed down in numerous manuscripts, and from the last quarter of the 15th century, also in many printed documents. Curiously, the latter are in the name of Canutus, Kami(n)tus, and the like. Virtually all the prints are undated, and were usually published without an indication of the place of publication or the name of the printer. The editions from the Southern Netherlands are no exception to this general rule. After a very brief description of the author and the contents, some attention is paid to the distribution of the publication in Europe, followed by a more detailed discussion of the different editions in Antwerp, published by Mathias van der Goes and Gheraert Leeu. The bibliographical analysis of these prints made wide use of the results of bibliographical research, and of the incomparable aid of the IISTC, the bibliographical data bank of incunabula. Thus the presumed dates can be compared with the latest results, and in this way be confirmed, altered or stated more precisely. Finally, following this bibliographical exercise, the author examines whether these dates could conflict with what we know about the plague epidemic in our country from other sources. PMID- 10379206 TI - [Measures taken against the plague in Diest in the fifteenth and sixteenth centuries]. AB - Older literature about the city of Diest reveals that in 1348 the plague wiped out half of the population; in 1439, 1523 and 1578-1579, the plague struck again. In the course of time the municipal authorities issued ordinances which were aimed at combating the disease and stopping it spreading. These ordinances were first published and then renewed or modified several times. This was done in the years 1469, 1483 and 1519. These texts have virtually the same contents and all contain about twenty articles of a prophylactic nature. A house where people had died of the plague had to remain shut for a number of months, and the relatives were not allowed in public for a while. Dirty water could not be emptied into the gutters, food had to be placed by the door, the clothes of those who were sick or who had died could not be washed in wells or in the river Demer and could not be sold, people caring for the sick had to wear a white sign, no one was allowed to take in any sick person from outside Diest, etc. Other articles are about refuse in the streets and pigs roaming around freely. Infringements were made punishable by fines, or if the perpetrators were insolvent, they had to make a pilgrimage to Strasbourg, Cologne or Rome. The ordinance of 1469 was issued again in 1472. It seems to be based on an ordinance of the city of Louvain, as this indication of the origin in the text was replaced by the word "Diest", and references to places in Diest were added. The ordinance of 1519 was confirmed in 1523, 1532, 1543 1544, 1558, 1574 and 1579. Separate regulations were issued in 1530, 1532-1533, 1573, 1578-1579, 1599. They relate to infected clothes and household articles, dung heaps, dead animals in the Demer, people who came into contact with anyone suffering from the plague, epidemics in the area, such as Beringen (1556) and Turnhout (1571). Diest also had municipal plague masters; these were surgeons. A number of names and data are known to us from municipal accounts from 1516 onward. Diest also had a plague house. From 1470, the Alexians and Black Sisters worked there, joined later by the Grey Sisters. The last sort of prophylactic measures against the plague were of a religious nature. These were the general processions in which the guilds had to participate, as well as one person from every household. Finally, severe fines were imposed for cursing and swearing. PMID- 10379207 TI - [Joannes de Vesalia, De Epidemia]. AB - The plague treatise by Joannes de Vesalia (ca. 1401-ca. 1476) is the subject of this article. The author plants to publish an edition and translation. In this contribution he describes the existing manuscripts (Stresa, Centro Internazionale di Studi Rosminiani, ASIC A2 21, and Rome, Biblioteca Apostolica Vaticana, Reg. lat. 1450), reconstructs their history and discusses their importance for reestablishing the original text. On the other hand, the author makes some general remarks on the content of the treatise. PMID- 10379208 TI - [Thomas Montanus, author of the plague treatise, Qualitas loimodea sive pestis Brugana: some biographical data]. AB - Thomas Montanus (1617-1685), physician of the City of Bruges, is the author of a voluminous treatise on plague, published in 1669, and written after an epidemic of plague in Bruges in 1666. A translation in Dutch of the treatise will be published shortly. In this contribution some biographical data on Montanus are presented: his descendence, his education, his career and his professional experience with plague. On the other hand the structure of the plague treatise Qualitas loimodea sive pestis Brugana is briefly mentioned. PMID- 10379209 TI - [Comparative study of the ideas about causes, disease mechanisms and therapies of the plague on the basis of the plague treatises of the Medical Faculty of Paris (1348-1349), of Joannes de Vesalia (after 1454) and of Thomas Montanus (1669)]. AB - The comparison between the consilium of the Faculty of Paris (14th century), and the treatises by Joannes de Vesalia (15th century) and by Thomas Montanus (17th century) shows that the concepts with regard to the causes and the mechanisms of and the proposed preventive and curative measures against the plague did in essence not change for over three centuries. In fact this is not very surprising, since the development of modern physiology and physiopathology only started gradually in the second half of the 19th century. Furthermore, the plague bacillus has only been identified in 1894. However, the preventive and curative measures against plague that are proposed in the three treatises, do not result from an uncontrolled imagination. Most of these measures have a rational basis and are the result of--although erroneous--concepts with regard to the causes and mechanisms of the disease. PMID- 10379210 TI - [Comparative study of the medicines used in the fight against plague on the basis of the plague treatises of the Medical Faculty of Paris (1348-1349), of Joannes de Vesalia (after 1454) and of Thomas Montanus (1669)]. AB - The remedies proposed for the prevention and treatment of the plague by the Medical Faculty of Paris and by Joannes de Vesalia are mainly derived from plants and animals apart from some minerals used in medieval medicine. Alchemical preparations, absent in the Compendium, are rarely mentioned by Joannes de Vesalia. About 90% of the simples preconized by the Faculty of Paris are still used as remedies in the tractates of Joannes de Vesalia and Montanus. The development of chemistry in the 16th and 17th centuries is responsible for the introduction of 'chemical' medicines in therapy. Montanus accepts these remedies with some reserve but favours also amulets and magic drugs. The plethora of medicines proposed demonstrates the inefficacy of therapeutics. PMID- 10379211 TI - [Attempts to inoculate against plague in the eighteenth and nineteenth centuries]. AB - In the middle of the 18th century, inoculation against smallpox became more and more common, and attempts were also made to test the same principle, viz. inoculation with the agents causing the disease for other human and animal diseases. It was tried for rinderpest, measles and sheep pox. In addition, there were some suggestions for using the principle against the plague. The disease had disappeared from Western Europe by this time, but still raged in eastern countries, such as Russia. However, the government rejected the proposal for trial inoculations in Moscow. During the first half of the 19th century, the plague was still widespread in the Middle East, where different European doctors worked on combatting it. The first documented inoculation trial was carried out by a certain Mr. Whyte, an English physician who inoculated himself and four assistants in 1801. All five died a few days later. In the following years, more tests were carried out, inter alia: in 1802, by Desgenettes, the chief physician of the French army in the Middle East; in 1803, by Eusebio Valli, an Italian physician in Constantinople; in 1818 and 1819 by Sola, a Spanish physician in Tangier. However, none of these tests produced clear results. During the epidemic in Egypt in the 1830s, further inoculation tests were carried out by a group of French plague specialists with the main aim of establishing whether the plague could be transmitted between humans. These tests did not result in any clear conclusions either. Following the discovery of the plague bacillus at the end of the 19th century, a number of different live and dead vaccines were developed, and were also used in endemic areas, but the level of efficiency has never become very clear. This is not really surprising, as even the disease itself often does not provide strong immunity, and reinfections are by no means uncommon. PMID- 10379212 TI - [The plague and the art of painting: some examples]. AB - The author gives four examples of pictures representing the plague: "The Plague of Asdod" by Nicolas Poussin, a picture of Saint Roch by the Master of Frankfort, "Saint Roch Interceding for the Plague-stricken" by Peter Paul Rubens and "The Sisters of Saint Elisabeth Hospital in Antwerp and the Works of Mercy" by Jacob Jordaens. PMID- 10379213 TI - Rethinking case reports. PMID- 10379215 TI - The power of herbs. PMID- 10379214 TI - Medical futility. PMID- 10379216 TI - Is herpes zoster unilateral? PMID- 10379217 TI - Heart failure: Part I. First hospitalization and post-hospital care. PMID- 10379218 TI - Recurrent cystitis in nonpregnant women. AB - Consistent evidence from RCTs shows that antibiotic prophylaxis (either continuous or postcoital), using trimethoprim TMP-SMZ, nitrofurantoin, or a quinolone, reduces infection rates in women with high rates of recurrent cystitis (at least two per year). Limited evidence suggests that intermittent patient administered treatment (taken at the onset of symptoms) is an effective alternative management strategy to continuous antibiotic prophylaxis in women with high rates of infection (at least two per year). Limited evidence suggests that long-term prophylaxis is likely to benefit women with a baseline rate of more than two infections per year over many years. However, long-term treatment has not yet been evaluated in RCTs. In women who experience recurrent, uncomplicated cystitis, there is no evidence to support routine investigation of the urinary tract with excretory urography, ultrasonography, cystoscopy, or voiding cystourethrography. No specific subgroups of women who would clearly benefit from investigation have yet been adequately defined. PMID- 10379219 TI - Treatment advances in rheumatoid arthritis. PMID- 10379220 TI - Is bungee jumping safe? PMID- 10379221 TI - What advice can I give my patients with lymphedema? PMID- 10379222 TI - Taking a sexual history. PMID- 10379223 TI - Nonbeneficial or futile medical treatment: conflict resolution guidelines for the San Francisco Bay area. Bay Area Network of Ethics Committees (BANEC) Nonbeneficial Treatment Working Group. PMID- 10379224 TI - Therapeutic effectiveness and social context: the case of lobotomy in a California state hospital, 1947-1954. PMID- 10379225 TI - Recent developments in medical care of Jehovah's Witnesses. PMID- 10379226 TI - Media watch. AB - In late 1997, Sharon Bernstein, a 35-year-old Los Angeles Times journalist and a new mother, was assigned the county hospital beat. Recently pregnant, the reporter was drawn towards stories of maternal and fetal health. So, she decided to look into obstetric malpractice claims against county hospitals. What she uncovered would change county hospital policy, lead to an assembly bill, and rekindle the medical debate about the safety of lowering Caesarean section (C section) rates. PMID- 10379227 TI - New malaria vaccine shows promise. PMID- 10379228 TI - Epidural analgesia must be paid for in cash in advance. PMID- 10379229 TI - FDA proposes testing animal feed antibiotics. PMID- 10379230 TI - American trade union aims to recruit more doctors. PMID- 10379231 TI - WHO Expert Committee On Drug Dependence. AB - Since its first meeting in 1949, the WHO Expert Committee on Drug Dependence, which consists of a group of internationally recognized authorities on the medical and pharmaceutical aspects of psychoactive substances, has played a central role in the global drug control system. The Committee has regularly recommended the classification of psychoactive substances according to the drug control conventions in force at different times, conventions which the Committee has typically helped to formulate. In this report, the Committee reviews three previously identified substances for inclusion in the drug control schedules and makes a preliminary analysis of other substances that may be reviewed in the future. The report also makes recommendations concerning a proposal of the Government of Spain to extend control collectively to isomers, esters, ethers and pharmacological analogues of psychotropic substances under control. The Committee further examines the feasibility of controlling tobacco under the existing drug control conventions. PMID- 10379232 TI - Epidemiologic, pathogenic and molecular analysis of recent encephalomyocarditis outbreaks in Belgium. AB - In 1991 EMCV was isolated for the first time in Belgium from the offspring of a sow with reproductive failure. From August 1995 until December 1996, EMCV was diagnosed in 154 Belgian pig holdings in association with myocardial failure and sudden death in fatteners and suckling piglets or with reproductive failure in sows. To clarify some epidemiological aspects 3 EMCV isolates characteristic for the different clinical pictures and outbreaks were studied. Field observations and animal experiments indicated that the pathogenicity induced by each isolate is specific for one age category and that the spread of the virus is limited. The presented data also suggest that rodents may play a role in the transmission of EMCV but that pig-to-pig transmission is at least as important. Molecular analysis of two separate regions on the genomes of the respective EMCV isolates showed that the 1995-96 EMCV epizootic in Belgium was due to a new virus introduction. Furthermore, the VP1 coding gene is proposed as a marker of virulence. PMID- 10379233 TI - The effect of buparvaquone treatment on the levels of some antioxidant vitamins, lipid peroxidation and glutathione peroxidase in cattle with theileriosis. AB - Plasma levels of vitamins A, E, beta carotene, both plasma and erythrocyte glutathione peroxidase (GSHPx), lipid peroxidation (LPO) and reduced glutathione (GSH) were investigated in cattle naturally infected with Theileria annulata and treated with buparvaquone. There were two groups each containing 30 cattle. Naturally infected cattle were used in the second group. Buparvaquone (2.5 mg/kg body weight) was administered to animals in the second group. Blood samples were taken from control animals, and immediately before treatment, and from animals 10 days after the injection of buparvaquone. Detection of the infected animals was carried out by blood smears. Plasma vitamins A, E, beta carotene, both plasma and erythrocyte GSHPx, LPO and GSH levels were determined. The levels of LPO in plasma and erythrocyte samples were significantly (P < 0.05, P < 0.01) higher after treatment than in either control animals or before treatment. Plasma levels of antioxidant vitamins, vitamin E and beta carotene were significantly (P < 0.05, P < 0.01) lower after treatment than in either control animals or before treatment, while the vitamin E level was found to be higher before treatment than in either the control group or animals after treatment (P < 0.05, P < 0.01). The levels of vitamin A in plasma and the activity of GSHPx and GSH in both plasma and erythrocytes in control animals after and before treatment did not differ significantly. In conclusion, we observed that there was a decreased plasma level of vitamin E and beta carotene and an increased level of LPO in cattle treated with buparvaquone. Buparvaquone might function in the treatment of Theileria annulata by forming free radicals. PMID- 10379234 TI - Serotypes of Pasteurella haemolytica and Pasteurella trehalosi isolated from farm animals in Hungary. AB - The biochemical and serological characteristics of 486 P. haemolytica and 31 P. trehalosi strains (517 in total) isolated from different lesions of cattle, sheep, goats, pigs and poultry were examined. A total of 476 P. haemolytica strains (97.9%) showed the characteristics typical of the former biotype A of P. haemolytica, while 10 isolates (2.1%), all from poultry, could not be biotyped. A total of 481 strains (93.0%) could be assigned to one of the 17 serotypes of P. haemolytica-P. trehalosi and 36 strains (7.0%) could not. The majority (83.6%) of the cattle isolates were serotypes A1 and A2. Among strains isolated from sheep all serotypes of P. haemolytica could be identified with the exception of A14, but serotypes A1, A2, A6, A8 and A5 were the most frequent. The overwhelming majority (94%) of the caprine isolates were A2, other serotypes occurred only sporadically. The pig isolates, which could be isolated only very rarely, represented different serotypes, while none of the 10 strains isolated from poultry could be biotyped or serotyped. PMID- 10379235 TI - Experimental infections with Actinobacillus pleuropneumoniae in pigs--I. Comparison of five different parenteral antibiotic treatments. AB - SPF pigs aged 10 weeks were infected intranasally with Actinobacillus pleuropneumoniae serotype 2. After the onset of clinical symptoms of respiratory disease, which occurred 20 h post-infection, parenteral treatment with ceftiofur, danofloxacin, enrofloxacin, penicillin or tiamulin was initiated (n = 8 per group). Untreated groups, of which one was infected, served as controls. The uninfected control group did not show any signs of disease, while the infected control group was severely affected by the infection and also expressed a decreased weight gain following the challenge. Based on clinical signs, the magnitude of pathological lesions in the respiratory tract found at necropsy performed 17 days post-infection and the number of reisolates of A. pleuropneumoniae made at necropsy, treatments with the quinolones (danofloxacin and enrofloxacin) and the cephalosporine (ceftiofur) were superior to those with penicillin and tiamulin. The latter groups also developed antibodies to A. pleuropneumoniae to a larger extent. Some of the pigs treated with ceftiofur and danofloxacin developed antibodies to A. pleuropneumoniae, and the microbe was reisolated from approximately 50% of these animals. In contrast, pigs treated with enrofloxacin did not develop antibodies to A. pleuropneumoniae, and the challenge strain was not found at necropsy. The performance with respect to daily weight gain and feed conversion corresponded well with the clinical signs developed and the findings made at necropsy. The decreased growth recorded during the acute phase of the disease was, to a large extent, caused by a reduced feed intake. PMID- 10379236 TI - Experimental infections with Actinobacillus pleuropneumoniae in pigs--II. Comparison of antibiotics for oral strategic treatment. AB - The present study was aimed at scrutinizing the efficacy of oral antimicrobial treatments at experimental challenge using a strain of Actinobacillus pleuropneumoniae serotype 2 known to cause severe disease. SPF pigs aged 10 weeks were infected intranasally and the antimicrobial treatments were initiated 5 h prior to that exposure. Several antimicrobial drugs, as well as the length of the treatment period, were elucidated. The outcome of the challenge was monitored by registration of clinical symptoms, weight gains and the development of serum antibodies to A. pleuropneumoniae. At necropsy, the magnitude of pathological lesions in the respiratory tract and the rate of reisolation of the infective strain were recorded. Animals that became diseased displayed a decreased growth rate caused, to a large extent, by a reduced feed intake. The performance with respect to daily weight gain and feed conversion corresponded well with the clinical signs developed and serologic reactions, as well as with the findings made at necropsy. The results obtained among pigs treated with enrofloxacin, but also with florfenicol or chlortetracycline, were superior to those of pigs treated with penicillin, tiamulin or tilmicosin. A positive effect was obtained using a strategic in-feed medication against infection with A. pleuropneumoniae. Provided that the drug used is effective against the target microbe, initiating treatment prior to infection appeared to be more important than the length of the treatment. It should, however, be remembered that A. pleuropneumoniae was reisolated from all but one medicated group following an experimental challenge given after initiating the medication. Consequently medical treatment as described did not eradicate the microbe. PMID- 10379237 TI - Experimental infection by Vibrio anguillarum in mice and guinea pigs. AB - The fish pathogen Vibrio anguillarum causes a lethal infection in farmed fish characterized by hemorrhagic septicemia. There are no reports of experimental laboratory infections in warm-blooded animals. We investigated the effects of an intraperitoneal infection with different doses of a V. anguillarum suspension in mice and guinea pigs. The infection caused a 95-100% of mortality in 24-48 h. Hemorrhagic septicemia was observed at necropsy and confirmed by histological and hematological examination. Immunohistochemically positive bacterial clumps were detected exclusively in vessel lumen in all examined organs, including brain, and V. anguillarum was reisolated in pure culture from all organs, particularly from the kidney. Blood analysis showed erythropenia and leukopenia with granulocytosis in mice, platelet reduction and leukopenia with lymphocytosis in guinea pigs. PMID- 10379238 TI - Comparison of the dot-immunobinding assay with the serum agglutination test, the rose bengal plate test and the milk ring test for the detection of Brucella antibodies in bovine sera and milk. AB - In this study, Brucella antibodies in bovine sera and milk were detected using the dot-immunobinding assay (DIA), the serum agglutination test (SAT), the Rose Bengal plate test (RBPT) and the milk ring test (MRT). For this purpose, a total of 116 paired blood and milk samples collected at the same time from 56 aborted and from 60 healthy dairy cows was examined. In DIA, a nitrocellulose membrane (NCM) was used as the solid phase. Antigen adsorbed on the NCM was extracted from Brucella abortus S99 by heat treatment. The results obtained by DIA were compared with those of SAT, RBPT and MRT. Of the 116 paired blood and milk samples, 24 were positive and 72 were negative by all tests used. Serum samples of six aborted cows were positive by DIA, SAT and RBPT but the milk samples were negative by DIA and MRT. Serum and milk samples of four aborted cows gave positive reaction only by DIA tests. The remaining six aborted cows were negative only by MRT and two of them were negative by both RBPT and MRT. Four sera of healthy cows were found to be positive only by SAT. PMID- 10379239 TI - The purpose and legacy of private care. PMID- 10379240 TI - Cosmetic dentistry 99. PMID- 10379241 TI - Analyzing the 1998 fee survey. PMID- 10379243 TI - Why is esthetic dentistry grouped with the outlaws? PMID- 10379242 TI - The art of setting fees. Interview by Dr. Joseph A. Blaes. PMID- 10379244 TI - Keeping honest people honest. Embezzlement in the dental office. PMID- 10379245 TI - Murder by proxy: death of a disability claim. PMID- 10379246 TI - The value of value. PMID- 10379247 TI - Raises. How to do the right thing. PMID- 10379249 TI - Dental web sites that work. PMID- 10379248 TI - San Diego dentists win battle for intraoral camera tax credit. PMID- 10379251 TI - Viscoelastic stress analysis of thermally compatible and incompatible metal ceramic systems. AB - OBJECTIVE: The purpose of this study was to analyze transient and residual midpoint deflections and stresses in metal-opaque porcelain-body porcelain systems with matched and mismatched thermal contraction coefficients. METHODS: Calculations and measurements were made for seven trimaterial strips that covered a wide range of thermal contraction mismatches among constituent materials. Midpoint deflections were measured in a beam-bending viscometer during slow cooling from an initial temperature of 700 degrees C. Linear regression analysis with a correlation coefficient of 0.950 was used to compare measured and calculated residual midpoint deflections. Stress relaxation data were fit to a three-term exponential series by nonlinear regression analyses with correlation ratios ranging from 0.9972 to 0.9999. RESULTS: While finite element analyses correctly predicted the general shape of the deflection behavior as a function of temperature for all combinations, the best agreement between measured mean residual midpoint deflections and calculated values (+250 microns vs. +268 microns) was obtained for strips composed of a Au-Pd alloy (alpha m = 13.5 ppm/ degree C) with a medium expansion opaque porcelain (alpha o = 13.3 ppm/degree C) and a high expansion body porcelain (alpha B = 14.4 ppm/degree C). The highest calculated residual tensile stress of +26 MPa at the surface of body porcelain was associated with the 0.5-mm-thick Ni-Cr-Be alloy strip (alpha m = 15.1 ppm/degree C) with medium expansion porcelains (alpha o = 13.5 ppm/degree C and alpha B = 13.9 ppm/degree C). The smallest measured residual deflection (+10 microns) was also associated with this combination. The results of this study indicated that metal-ceramic strips are sensitive indicators of stress development caused by a thermal contraction mismatch; however, the magnitudes of the residual deflections do not necessarily correlate with the stress magnitudes in the ceramic. SIGNIFICANCE: Currently there are no U.S. or international standards that define the maximum difference in thermal contraction coefficients that can exist between a metal and its ceramic veneer without causing transient failures of ceramic during cooling or delayed failures in ceramic because of high residual tensile stresses. The present research represents a major step in understanding the various factors that influence the development of transient and residual stresses. A knowledge of the effects of process variables on stress development is necessary for selection of potentially successful metal-ceramic systems and for optimizing the design of dental prostheses. PMID- 10379250 TI - Eugenol diffusion through dentin related to dentin hydraulic conductance. AB - OBJECTIVES: The purpose of this study was (1) to find an easy way of evaluating the concentration of eugenol in cell culture fluids; (2) to confirm the relationship between the concentration and the cytotoxicity of eugenol in vitro; (3) to evaluate the cytotoxicity of four temporary eugenol-based filling materials: IRM, super EBA, Kalsogen and zinc oxide-eugenol cement; and (4) to establish a relationship between dentin permeability, eugenol diffusion and cytotoxicity. METHODS: (1) The concentration of eugenol was measured with a spectrofluorimeter; (2) the cell viability of L 929 cells cultivated for 24 h with eugenol-containing medium was evaluated by the MTT assay; (3) after measurement of hydraulic conductance, occlusal cavities in human teeth in vitro were filled with the restorative materials. The cytotoxicity was measured with undiluted test medium and with various dilutions in culture medium; (4) after Lp measurement, the eugenol concentration in the media in the pulp chamber that diffused from IRM and 10(3) mol/l eugenol solution was measured. RESULTS: (1) A proportional relationship (p = 0.001 and r = 1) was found between the concentration of eugenol; (2) eugenol started to be cytotoxic at 10(-5) mol/l and killed 95% of the cells at 10(-3) mol/l; (3) zinc oxide-eugenol cement was the most cytotoxic filling material when tested with the 1:100 dilution; (4) a significant relationship was found between Lp and cytotoxicity (p = 0.04) depending on the dilution of the test medium. A significant relationship was found between Lp and eugenol diffusion from a 10(-3) mol/l solution (p = 0.03) but not between Lp and eugenol diffusing from solid IRM (non significant). SIGNIFICANCE: Eugenol diffusion from zinc oxide-eugenol cement appears to depend more on the role of hydrolysis of eugenol from zinc oxide-eugenol cement than on dentin permeability. PMID- 10379252 TI - Comparison of three fracture toughness testing techniques using a dental glass and a dental ceramic. AB - OBJECTIVES: Various methods aimed at determining the fracture toughness of ceramics in mode I (KIc) have been described in the literature. The accuracy, scatter and the interexaminer reproducibility of KIc depend strongly on the procedural approach, the test parameters used and the conditioning of the specimens. The purpose of the present study was to compare fracture toughness values obtained using two indentation methods as well as a newly established fracture mechanics test. METHODS: The following methods for KIc determination were applied: (1) indentation fracture (IF), (2) indentation strength (IS) and (3) the single-edge-V-notched-beam test (SEVNB). The materials tested were a low fusing dental glass (Duceram LFC) and a feldspar-based porcelain (IPS classic). Data were compared by ANOVA and Tukey's multiple comparison test (p < or = 0.05). RESULTS: For both materials, KIc coefficients of variation ranged between 10 and 14% for IF and 7 and 10% for IS. The IS technique demonstrated a load dependency for the IPS porcelain which was not observed when using the IF method. The SEVNB test provided consistent results with coefficients of variation between 1 and 3%. SEVNB toughness values for the IPS porcelain were in agreement with the IS technique. However, halfpenny shaped cracks were observed at the tip of the notch of all LFC specimens thus leading to underestimated KIc values. SIGNIFICANCE: The overall aim of this type of study is to select testing procedures that are as expedient and reliable as possible. This study has shown that all three methods agreed within 10%. However none of the procedures proved absolutely straightforward. Decision on which method to use should be based on a sound understanding of the conceptual limitations and technical difficulties inherent to each technique. PMID- 10379253 TI - Reduction of composite contraction stress through non-bonded microfiller particles. AB - OBJECTIVES: To determine the reduction in composite polymerization stress through the addition of non-bonded microfiller particles. METHODS: Microfillers that were unsilanated, silanated, and treated with a nonfunctional silane were added to dental resin and to a small-particle composite. The contraction stress generated by these materials was measured by polymerizing them between glass plates mounted in a mechanical testing machine. The maximum force was recorded 15 min after photo-initiation. Results were analysed by ANOVA (analysis of variance)/Turkey's test (p < or = 0.05). RESULTS: The addition of non-functional silanated microfillers to dental resin resulted in a significant 50% decrease in polymerization stress. The addition of unsilanated microfillers did not reduce the contraction stress. When added to small-particle composite, the unsilanated microfillers produced a significant 30% reduction in contraction stress compared to the composite containing silanated microfillers. The non-functional silanated microfillers did not reduce the contraction stress in the small-particle composite. SIGNIFICANCE: The polymerization shrinkage of dental composite can impose high levels of stress on the tooth surfaces to which it is bonding. This contraction stress can lead to failure of bond formation with the surrounding tooth structure. Microfiller particles that are not bound to the resin matrix might provide sites for relief of internal stresses, significantly reducing contraction stress in dental composite. PMID- 10379254 TI - Effect of acetic acid on the fluoride release profiles of restorative glass ionomer cements. AB - OBJECTIVES: This study investigates the fluoride release of glass ionomers in an acetic acid solution in order to substantiate a model according to which the short-term release results from an elution of loosely bound fluoride and the long term release from an erosive leaching of the glass particles in the bulk of the cement. METHODS: Individual fluoride release profiles of five specimens of 10 acid-base setting restorative glass ionomers were obtained by determining the amounts of fluoride released by each sample at 37 degrees C in consecutive elutions for up to 140 days with 25 ml of a 0.01 mol/l acetic acid solution with pH = 4. Differences in the fluoride release profiles were determined with a Multivariate Data Analysis on the basis of a Principal Component Analysis. RESULTS: The fluoride release profiles of the 10 glass ionomers can be classified into five distinct groups which are characterized by a cumulative fluoride release described by the equation [F]c = [F]l t/(t + t1/2) + beta square root of t. The parameters ([F]l, t1/2) and beta are characteristic for the materials in the groups, and refer to the short-term and long-term fluoride release, respectively. The acidic solution enhances both processes compared to an elution in water, the effect being more pronounced for the long-term release. SIGNIFICANCE: The fluoride release mechanism is intrinsically the same as determined for elutions in water. The increased amount of fluoride released under acidic conditions, especially in the long term, corroborates that an erosive leaching of the glass particles in the bulk of the cement accounts for the long term fluoride release. PMID- 10379255 TI - The effect of stiffness on the localization of tensile stress at the surface of bonded posterior restorations. AB - OBJECTIVES: The aim of this study is to assess the distribution of tensile stress under the conditions representative of tooth-food contact in a restored posterior tooth. The ideal stiffness for a bonded restoration will be predicted. METHODS: A two-dimensional plane-strain finite element representation of a restored first maxillary molar is created. The restoration with Class I/II bucco-lingual geometry is assigned a range of Young's moduli (E = 10-80 GPa), representative of the range of materials available. In addition, a hypothetical multi-phase model, in which stiffness is gradually increased from the intercuspal concavity to the adjacent enamel, is also created. A food particle is modeled and pushed close to the interface onto the site of (1) intact enamel and (2) restored surface. RESULTS: For all single-phase models tested, the magnitude of tensile stress at the surface is far greater than that present elsewhere in the tooth. However, the exact position and magnitude of the maximum tensile stress varied according to the modulus assigned to the restoration and the position of load. The results of the model imply that interfacial problems are likely in low-modulus restorations (E = 10-20 GPa), whereas stress-related problems could occur in the intercuspal concavity of high-modulus restorations (E = 40-80 GPa). Of all the single-phase models, tensile stresses are lowest when the Young's modulus assigned to the restoration is 30 GPa. These tensile stresses are reduced in the multi-phase model. SIGNIFICANCE: Given the limitations of the model, the results indicate that the most suitable modulus for a single-phase posterior bonded restoration is around 30 GPa. Such a modulus is approached in compact-filled composites. In addition, the highly desirable dissipation of load in the multi-phase model should warrant further investigation. It is suggested that the use of an incremental filling technique with region-specific proportions of hard-filler could be one way forward. PMID- 10379256 TI - Thermal stresses in metal-ceramic specimens for the ISO crack initiation test (three-point flexure bond test). AB - OBJECTIVES: At the October 1996 meeting of the ISO/TC106/SC 2/WG 1 working group, a special three-point flexure test for the characterization of the metal-ceramic bond was incorporated in the standard (ISO CD 9693). Due to the fabrication process, like real porcelain-fused-to-metal (PFM) restorations, the specimens contain thermal (eigen-) stresses upon which the load stresses are superimposed in the actual test. This study is devoted to the determination of these residual thermal stresses. METHODS: The residual thermal stresses in the specimen were calculated with the aid of the finite element method (FEM) using an especially fine mesh in the vicinity of the edge of initial debonding. Young's modulus, EM, of the alloy was varied within the interval 80 GPa < or = EM < or = 220 GPa which covers the spectrum of dental alloys. The analysis also allows the calculation of thermal stresses as a function of the difference delta alpha = alpha M - alpha C of the coefficients of thermal expansion of alloy and ceramic and the glass transition temperature theta G of porcelain. RESULTS: The thermal shear and normal stresses at the bond interface concentrate at the end of the ceramic veneer and practically vanish over about three quarters of the central part of the layer. The larger the Young's modulus, EM, of the alloy, the higher both stresses. SIGNIFICANCE: The results permit a deeper comprehension of the debonding process in the test: shear stress induced by loading increases the overall shear stress at the end of the bond interface, whereas load tensile stress is buffered by thermal compressive stress. PMID- 10379257 TI - Effect of the application of dentin primers and a dentin bonding agent on the adhesion between the resin-modified glass-ionomer cement and dentin. AB - OBJECTIVES: This study was designed to evaluate the influence of the application of dentin primer and/or dentin bonding agent on the adhesion of a resin-modified glass-ionomer cement to dentin. METHODS: Bovine dentin was pretreated with Dentin Conditioner or EDTA 3-2 solution, primed by an experimental dentin primer, and applied with a dentin bonding agent. A resin-modified glass-ionomer cement, Fuji II LC, was then adhered to the dentin. The tensile bond strength between the light-cured glass-ionomer cement and the pretreated dentin was measured. The components of the experimental dentin primers were 2-hydroxyethyl methacrylate (HEMA), glyceryl methacrylate (GM) and a water-soluble photo-polymerization initiator, 2-hydroxy-3-(3,4-dimethyl-9-oxo-9H-thioxanthen-2-yloxy)-N,N, N trimethyl-1- propanaminium chloride (OTX). Significant differences in the data were examined by an analysis of variance and Scheffe's test for multiple comparisons between the means at p = 0.05. RESULTS: A significantly higher mean bond strength between the Fuji II LC and dentin was obtained by EDTA 3-2 pretreatment, QTX/GM priming, and LB Bond application. This value was comparable with that obtained with the resin composite system. Scanning electron microscopy observation showed the formation of a hybrid layer with a thickness of 1-1.5 microns. SIGNIFICANCE: The data obtained in this investigation suggest that the adhesion of Fuji II LC to dentin is closer to that provided by a resin composite system than to that of conventional glass-ionomer cements. PMID- 10379258 TI - Fracture toughness determination of composite resin and dentin/composite resin adhesive interfaces by laboratory testing and finite element models. AB - OBJECTIVES: The reliability and validity of the adhesive bond toughness of dentin/composite resin interfaces were studied from the standpoint of fracture mechanics. METHODS: The fracture toughness (KIC) and fracture energy (JIC) values of two different composite resins (Brilliant Dentin and P50) were determined by using single edge notch (SEN) specimens loaded in three point bending and the results were analyzed by the t-test method (p < 0.1). The fracture loads of dentin/composite resin interface with different initial crack lengths were obtained experimentally. The adhesive fracture energy (J(adh)), residual fracture energy (J(res)) and effective (total) fracture energy (J(eff)) for the symmetrical bimaterial (SBM) joint specimen for dentin/composite resin interfaces were calculated and the applied fracture energy (J(appl)) values under the mastication force were obtained for the axisymmetric tooth models. All numerical calculations were carried out by the finite element method and software programs were prepared according to fortran 77. RESULTS: The fracture toughness and energy values obtained experimentally for Brilliant Dentin were found to be higher than those for P50. It was seen that, calculated J values (J(adh) and J(res)++) changed with the crack length; but the effective fracture energy (J(eff)++) was independent of the crack length, as expected. The applied fracture energy (J(appl)) and effective fracture energy (J(eff)) are considerably smaller than the experimentally determined JIC values of composite resins. SIGNIFICANCE: The bonded interface tends to produce microscopic flaws which could act as critical stress risers promoting interfacial failures. The initiation and propagation of such flaws under the mastication forces can be followed by fracture toughness (KIC) or fracture energy (JIC) in linear elastic fracture mechanics (LEFM). PMID- 10379259 TI - Marginal adaptation and retention of a glass-ionomer, resin-modified glass ionomers and a polyacid-modified resin composite in cervical Class-V lesions. AB - OBJECTIVES: An 18-month follow-up clinical trial of one conventional glass ionomer (HIFI Master Palette), three resin-modified glass-ionomers (Fuji II LC, Vitremer, 3M Exp. 155) and one polyacid-modified resin composite (Dyract) was conducted to evaluate their clinical effectiveness in Class-V cervical lesions. In addition, the interface between dentin and two resin-modified glass-ionomers and one polyacid-modified resin composite was examined by scanning electron microscopy (SEM). METHODS: After evaluation of the restorations immediately following placement (baseline), all patients were subjected to a strict recall schedule with controls at 6, 12 and 18 months. The clinical effectiveness was recorded in terms of retention and marginal integrity, clinical microleakage, caries recurrence, and tooth vitality. A chi 2-test (p < 0.05) was used to test for significant differences between materials. In case of restoration loss or special defects, a replica was made to examine the surface texture and restoration margins by SEM. In vitro, the interface was examined by SEM after an argon-ion-beam etching technique was used to enhance surface relief and disclose interfacial substructures. RESULTS: Retention appeared to be good for all the materials tested. Marginal discrepancies were localized at the incisal enamel and/or the cervical dentin margin, except for the polyacid-modified resin composite that showed most of the defects at the incisal enamel margin. None of the systems could guarantee margins free of microleakage for a long time. In vitro, the type of dentin pre-treatment defines to a great extent the morphology of the resultant interface between dentin and the restorative material tested. SIGNIFICANCE: In this clinical study, the retention rate of the tested materials was good and even excellent for some products. Perfect marginal adaptation deteriorated too fast. The marginal adaptation of the polyacid-modified resin composite at the enamel site would probably have been better by the use of selective enamel or total acid etching. Marginal sealing remains a problem. Future research should concentrate on improving the marginal adaptation and sealing capacities before a broader clinical use can be advocated. PMID- 10379261 TI - Bond strength between cements and metals used for endodontic posts. AB - OBJECTIVES: Adhesive cements used with metal endodontic posts may decrease fracture in non-vital teeth. Results from studies that evaluate cements for post retention by pulling posts out of extracted teeth are difficult to interpret owing to the number of interfaces where fracture might occur. The objective of this study was to isolate the metal/cement interface for tensile bond strength testing and microscopic observation. METHODS: Three metals and seven cement treatments were examined for bond strength by using a truncated cone tensile test. The bond strength data were analyzed by a two-way ANOVA and Scheffe's multiple comparison test at p = 0.05. Specimens were examined at 50x magnification to determine the failure mode and with scanning electron microscopy (500x) to observe the surfaces after debonding. RESULTS: Significant differences in tensile bond strengths were found among cements compared within two of the metal groups. One of the metal groups had no significant differences among cement bonds. When comparing within cement treatment groups, two groups had significant differences in bond strength among the metals. Microscopic observations revealed adhesive, cohesive and mixed failure modes that varied with cement treatment and metal combination. The interaction between metal and cement was a critical determinant of the strength and characteristic fracture mode of the bond achieved. SIGNIFICANCE: Some of the cement treatments performed better (i.e., higher bond strength) with some metals than with others. Other cements had similar bond strengths with all three metals. Because of this interaction, careful consideration of the materials combination should help to maximize the bond at the metal/cement interface. PMID- 10379262 TI - Computer aided prediction and control of shrinkage porosity in titanium dental castings. AB - OBJECTIVES: The main objectives were to investigate the possibility and reliability of quantitative prediction and control of the concentrated shrinkage porosity (macroporosity) in titanium dental castings by means of a numerical simulation technique; and finally to optimize the filling and feeding system design for dental castings. METHODS: A commercial software, MAGMASOFT (Giessereitechnologie GmbH, Germany), was employed to simulate the mold filling and solidification process, and predict the shrinkage tendency in a sample dental casting, two simplified tooth crowns with a connector bar between them. The numerically predicted shrinkages were compared with the experimental results. The experiments were carried out on a centrifugal casting machine. The same geometric and processing parameters of the casting as in the simulations were strictly controlled. RESULTS: The computer predicted shrinkage porosity coincided with the performed experiments, demonstrating the reliability of the numerical model and the thermal physical data chosen for the calculations. Based on the above numerical model, several filling and feeding systems for the same casting were numerically simulated and compared. Finally an optimized design for this sample casting was proposed, and porosity-free castings were obtained. SIGNIFICANCE: It was expected that the numerical simulation technique could be further developed for dental laboratories to aid the real dental casting design. PMID- 10379260 TI - Adhesive bonding to dentin with iron (II) perchlorate primers and a tri-n butylborane-initiated luting agent. AB - OBJECTIVES: This study was conducted to measure the tensile bond strength of a resin to dentin when the dentin was primed with iron (II) perchlorate modified aqueous 2-hydroxyethyl methacrylate (HEMA) or an iron (II) perchlorate modified commercial self-etching primer (ED primer, Kuraray Co.). METHODS: Bovine dentin surfaces were ground flat and each specimen underwent one of the following two treatments: (1) priming with 2.0 x 10(-6) to 5.0 x 10(-4) mol/g iron (II) perchlorate in aqueous HEMA solutions after etching with 10 wt% phosphoric acid; (2) priming with self-etching primers containing 4.0 x 10(-7) to 2.0 x 10(-4) mol/g iron (II) perchlorate. Each specimen was then bonded to a stainless-steel rod with a luting agent (MMA-TBB resin) consisting of methyl methacrylate (MMA), poly(methyl methacrylate) (PMMA), and tri-n-butylborane (TBB) initiator. Tensile strengths of the bonded tooth specimens were then determined after 1 day immersion in water. Results were analyzed using ANOVA and Duncan's new multiple range test (p < 0.05). RESULTS: Tensile testing revealed the maximum mean bond strengths (22.5 MPa) when the dentin was primed with 2.0 x 10(-4) mol/g iron (II) perchlorate after etching with 10 wt% phosphoric acid aqueous solution. The highest level of bond strength with self-etching primer (19.6 MPa) was achieved using 1.0 x 10(-4) mol/g iron (II) perchlorate. SIGNIFICANCE: These bonding techniques, combining the use of iron (II) perchlorate modified HEMA primers with MMA-TBB resin, are potentially applicable for seating resin-bonded restorations. PMID- 10379263 TI - Micromorphological spectrum of acid-conditioned dentin following the application of a water-based adhesive. AB - OBJECTIVES: The goal of this study was to illustrate the micromorphological spectrum along the resin-dentin interface when a water-based, dentin adhesive (Scotchbond Multi-purpose) was applied to acid-conditioned dentin under different dry and wet bonding conditions. METHODS: Twenty-eight 1 mm dentin discs were each conditioned with 10% maleic acid for 15 s. Twenty-four of these discs were randomly divided into four groups, based upon the status of the remaining surface moisture: Group I (30 s dry); Group II (3 s dry); Group III (visibly moist) and Group IV (overwet). They were bonded using Scotchbond Multi-purpose. The remaining four discs were bonded using an experimental water-based primer containing 35 vol.% HEMA (Group V). Laminated dentin disc pairs were prepared for transmission electron microscopic examination. RESULTS: In all groups, diffusion of the polyalkenoic acid copolymer component of the primer into acid-conditioned dentin was localized to the surface region of the hybrid layer. The remaining part of the hybrid layer (the subsurface region) appeared variable. In Group I (30 s dry), collagen fibrils were collapsed. In Group II (3 s dry) and Group III (visibly moist), stained collagen fibrils were surrounded by wide electron-lucent interfibrillar spaces. In Group IV (overwet), a marked diffusion gradient was observed representing dilution of part of the primer components. In addition, an electron-dense primer phase, containing electron-lucent globular domains was invariably observed, irrespective of the hydration status of the demineralized collagen matrix. This electron dense phase was absent when HEMA alone was used as the primer (Group V). SIGNIFICANCE: With the use of a water-based adhesive, one could briefly air-dry the acid-conditioned dentin and allow the water in the primer to rehydrate the collapsed collagen matrix, without the risk of incomplete hybridization or tubular seal along the resin-dentin interface. However, care should be exercised to minimize dilution of the water-soluble primer component. PMID- 10379264 TI - Evaluation of metal ion release and corrosion resistance of ZrO2 thin coatings on the dental Co-Cr alloys. AB - OBJECTIVES: The aim of this study was to determine if electrolytic ZrO2 thin coatings increased the corrosion resistance and decreased the metal ion release of dental cobalt-chromium alloys. METHODS: Dental Co-Cr alloys were electrolytically deposited with ZrO2 ceramic coatings using a 0.0625 M ZrO(NO3)2 solution, at various potentials, for 500 s. The electrolytic ZrO2 gel-coated specimens were annealed at 723 K for 1 h in air. Scanning electron microscopy (SEM) was used to observe the morphology of the ZrO2 ceramic coatings on Co-Cr alloys. A dynamic polarization test was used to compare the corrosion resistance of the ZrO2 coated and uncoated Co-Cr alloys in artificial saliva. Metal ion concentrations were determined with graphite furnace atomic absorption spectroscopy (AAS). RESULTS: The SEM micrographs showed that the Co-Cr alloy can be coated with zirconia oxide at -0.7 V more homogeneously and more completely than at -1.5 V. The polarization curves indicated that the ZrO2 coating on Co-Cr alloys annealed at 723 K for 1 h in air exhibited better corrosion resistance in artificial saliva. The results of the AASs showed that the ZrO2-coated Co-Cr alloys decreased chromium ion release levels, as compared with the uncoated Co-Cr alloys. The scratch test indicated a good bond strength between the ZrO2 and Co Cr alloy. SIGNIFICANCE: The electrolytically deposited ZrO2 coatings on Co-Cr alloys may improve the corrosion resistance and decrease the release of metal ions. It is suggested that the electrolytic ZrO2 coating method could have a widespread application in dentistry in the future. PMID- 10379265 TI - The durability of adhesion to phosphoric acid etched, wet dentin substrates. AB - OBJECTIVE: The purpose of this study was to investigate the influence of remaining non-resin-impregnated, phosphoric acid demineralized dentin upon the long-term durability of specimens that were wet-bonded to bovine dentin substrates. METHODS: Prepared bovine dentin samples were etched with 65% phosphoric acid then rinsed with water and kept wet during application of 5 wt% 4 methacryloyloxyethyl trimellitate anhydride (4-META) in acetone primer. This was followed by application of a photocured dentin-bonding agent consisting of 4 methacryloyloxyethyl trimellitate anhydride/triethyleneglycol dimethacrylate camphorquinone/N-phenylglycine (4-META/TEGDMA-CQ/NPG). The tensile bond strength (TBS) of bonded specimens was determined after immersion in 37 degrees C water for various time intervals. Generated data were analyzed for statistical significance by one-way ANOVA and Duncan's New Multiple Range Test (p < 0.05). The dentin side of the tensile-load-fractured specimens was examined under optical and scanning electron microscopes (SEM). RESULTS: TBS decreased from 6.6 +/- 1.0 MPa after 1-day water immersion to 3.4 +/- 1.7 MPa after 1 month of water immersion. After 6 months of water immersion, TBS was found to be 3.9 +/- 0.9 MPa and this decreased to 2.0 +/- 1.0 MPa for specimens immersed in water for 1 year, a statistically significant difference (p < 0.05). Optical microscopic and SEM observations disclosed failure patterns within demineralized, non-resin impregnated dentin that increased with the period of water immersion. SIGNIFICANCE: The bond durability to wet dentin was poor when demineralized dentin was not resin-impregnated, resulting in exposure of collagen fibrils which hydrolyzed during long periods of water immersion. PMID- 10379266 TI - Ambient light working times of visible light-cured restorative materials. Does the ISO standard reflect clinical reality? AB - OBJECTIVES: The aim of this study was to determine whether a custom built light box could reliably reproduce clinical conditions and whether the current ISO standard test for the working time of visible light cured (VLC) materials (ISO. Dental Resin-based Restorative Materials. ISO:4049, 1991) reflected clinical reality. METHOD: The ISO test requires a VLC material to remain homogenous after exposure to an 8 kLux light source for 60 s. The relationship between the intensity of ambient lighting and the working times of visible light-cured (VLC) restorative materials was investigated by exposing 23 different VLC composite resin materials to a range of light intensities and measuring the working time in seconds: (a) under clinical operating conditions; (b) in a custom light box at 24 kLux; and (c) at 8 kLux in the light box. Working times were determined by placing 0.06 g samples between two glass plates and gently rotating until clefts or voids appeared. RESULTS: Clinical lighting conditions (24 kLux) gave a mean working time of 54 s (s.d. 14 s). In the light box at 24 kLux the mean working time was 54 s (s.d. 11 s). At 8 kLux (under otherwise identical conditions) the mean working time was 162 s (s.d. 49 s). SIGNIFICANCE: A 24 kLux illuminance in the laboratory gives an optimal correspondence to operative lighting conditions. This is three times the lighting level required for ISO purposes, and gives more realistic working times. PMID- 10379267 TI - Failure and fracture characteristics of glass poly(vinylphosphonate) cements. AB - OBJECTIVES: A glass poly(vinylphosphonate) cement, Diamond Carve, consisting of an ion leachable glass and a co-polymer of poly(vinylphosphonic-co-acrylic acid) was characterised. Samples were mixed for mechanical analysis using a linear elastic fracture mechanics (LEFM) approach. METHODS: Fracture toughness, flexural strength, Young's modulus, toughness and compressive strength were measured. Cement samples were tested at ageing times of 1, 7, 28, 84 and 168 days. RESULTS: Fracture toughness values in the range 0.6-0.82 MPa square root m were obtained. Young's modulus increased with ageing time from 8.6 GPa at one day to 14.3 GPa at 168 days. An increase in compressive strength from 149 to 242 MPa was also observed over the same time period. SIGNIFICANCE: The influence of ageing time had a significant effect on the mechanical properties of the cement. The expected rise in the mechanical properties was observed as a result of the ongoing crosslinking in the polysalt matrix. PMID- 10379268 TI - Cerium oxide as a silver decolorizer in dental porcelains. AB - OBJECTIVES: Silver-base alloys are known to produce an esthetically unpleasant yellow-green tint in the dental porcelain when making PFM restorations. Cerium oxide is used in dental porcelains to simulate the natural fluorescence found in human dental enamel, and has also been used in the glass industry as a decolorizer. The purpose of this study was to determine the effect of CeO2 additions on the resistance of dental porcelain to staining from silver contamination. METHODS: Five batches of porcelain were prepared according to Weinstein et al. (1962) with 0.00, 0.05, 0.10, 0.15 and 0.20 wt% additions of CeO2. To determine the resistance of these porcelains to silver staining, 0.10 wt% additions of the silver oxides were triturated into the prepared CeO2 porcelains prior to sample fabrication. This procedure provided a more quantitative method of staining than firing directly on silver alloys. Silver oxide was added in two valence states as Ag2O and AgO to test for any possible effects on staining. Samples were pressed into a 17 mm diameter mold, and fired to 960 degrees C under vacuum. Three additional samples were prepared from the non-cerium porcelain frit to produce a non-stained control group. Color measurements were made with a spectrophotometer on the ten experimental groups and the control group. The CIE L*a*b* color difference, delta E*, was calculated between the control and the experimental groups. RESULTS: There was a significant decrease in the silver staining of dental porcelains when CeO2 additions of 0.10 wt% or greater were used. SIGNIFICANCE: Cerium oxide additions in the range of 0.10 to 0.20 wt% caused a three-fold reduction in the staining of dental porcelain samples which had been doped with 0.10 wt% of AgO or Ag2O. PMID- 10379269 TI - Margin quality of titanium and high-gold inlays and onlays--a clinical study. AB - OBJECTIVES: Unalloyed titanium may be an economical substitute for gold alloys. The purpose of this study was to evaluate the suitability of unalloyed titanium as an alternative to gold alloys for posterior inlay and onlay restorations. METHODS: Fifty-four patients received 99 titanium restorations, 56 patients 96 gold alloy restorations. All titanium restorations were cast in a vacuum-pressure casting machine. Two weeks after insertion the clinical margin integrity was evaluated by replica technique. The maximum marginal gaps were assessed under the optical measuring microscope. The two tailed t-test for independent samples was used for statistical analysis. RESULTS: Accuracy of fit was significantly better in gold alloy than in titanium restorations. Mean maximum marginal gap between restoration margin and tooth structure were 72 +/- 18 microns (mean +/- S.D.) for titanium and 64 +/- 18 microns (mean +/- S.D.) for gold restorations. SIGNIFICANCE: Though marginal gap widths in titanium restorations did not yet match the gold standard, the data justify the use of titanium as an alternative to gold alloy for inlay and onlay restorations. PMID- 10379270 TI - Depth-resolved Raman microprobe examination of a commercial dental porcelain exposed to a simulated oral environment. AB - OBJECTIVES: The specific aims were investigation of the chemical transformations into the interior of a commercial dental porcelain upon exposure to a simulated oral environment and identification of structural alterations that contribute to the strengthening of the porcelain. METHODS: Samples of a commercial dental porcelain, Duceram LFC, were exposed to refluxing dilute acetic acid for 0, 1, 5, 10, and 72 h to simulate exposure to the oral environment. The control and refluxed samples were examined using a depth-resolved Raman microprobe system covering the low (0-2550 cm-1) and high (2000-4000 cm-1) frequency ranges. Spectra were processed using exploratory factor analysis. Significant factors were retained, accounting for 99.999% of the variance. RESULTS: The porcelain was found to be significantly depolymerized prior to acid-refluxing. Exposure to water and to acid resulted in the formation of a better-ordered region near the surface. In that region, three intermingled zones of chainlike metaborate and pyroborate, of boroxol rings, and of a mixture of borate and silicate tetrahedra were identified. The borate/silicate factor was dependent on refluxing time. Hydroxyl groups were detected in all of the samples, but no changes to the hydroxyl environment were noted until after 10 or more hours of refluxing. Then, molecular water and isolated hydroxyls/SiOH were detected. After 72 h, an additional unassigned factor around 2800-2950 cm-1 was noted. SIGNIFICANCE: Acid refluxing causes generation of ordered boron-containing species near the surface of the porcelain. PMID- 10379271 TI - Infraposition of ankylosed permanent maxillary incisors after replantation related to age and sex. AB - The extent of infraposition of replanted and subsequently ankylosed permanent incisors was examined in a longitudinal study of 52 patients. Study cast models were made during the follow-up period. Infraposition was evaluated on frontal photos of the study cast models taken parallel with the occlusal plane. The extent of infraposition was defined as the difference between the position of the incisal edge of the replanted incisor and the adjacent non-injured incisor in apico-coronal direction, measured with a digital caliper at 50x magnification of the negatives. The precision of this measuring procedure was 2.9% and the accuracy 2.0%. Marked infraposition was identified if the tooth was traumatized before the age of 16 in boys and before the age of 14 in girls. In addition, infraposition was observed when ankylosis developed in patients aged 20-30 years, with a yearly mean infraposition rate of 0.07 mm/year (range: 0.02-0.21 mm/year) in males and 0.07 mm/year (range: 0.00-0.12 mm/year) in females. The latter findings supported the concept of slow continuous eruption of the teeth. This phenomenon may have implications not only for the treatment of traumatized teeth but also for the treatment of tooth loss by osseointegrated implants, which represent an analogue to the ankylosed replanted tooth. PMID- 10379272 TI - Clinical experience of root canal filling by ultrasonic condensation of gutta percha. AB - Ninety human teeth with a total of 181 prepared root canals were filled with the use of ultrasonically energized spreaders as an aid for lateral condensation of gutta-percha cones. The results were evaluated on the basis of clinical and radiographic criteria. Adequate fillings were found in 93% of the canals and the overall success rate was 93%. All of the recalled patients were clinically comfortable. PMID- 10379273 TI - Dye leakage and SEM study of roots obturated with Thermafill and dentin bonding agent. AB - The apical seal of roots obturated with a dentin bonding agent and Thermafill with and without the use of sodium hypochlorite as an irrigating solution was compared by a dye leakage test. Roots obturated with Thermafill and a zinc oxide eugenol sealer were used as controls. Thirty-eight roots were prepared chemomechanically and divided into three experimental groups. The teeth of group 1 were filled with Thermafill and the dentin bonding agent using sodium hypochlorite as the irrigant. The teeth of group 2 were filled in the same way, but saline solution was used as the irrigant. Before the root canal was filled the smear layer was removed from the root canal walls of both groups by rinsing the root canal with a 17% EDTA solution. The teeth of group 3 were filled with Thermafill and a zinc oxide eugenol sealer. The teeth were immersed in 2% methylene blue solution. The root fillings of groups 1 and 2 leaked significantly more than those of group 3. The resin-dentin-guttapercha interface of group 1 was observed by scanning electron microscopy and showed a typical hybrid layer. An intimate contact between resin and dentin was present in group 2, but a resin dentin interdiffusion zone was only occasionally observed. The use of dental adhesives and the hybrid layer formation did not improve the seal of Thermafill root canal fillings. PMID- 10379274 TI - Number of roots and canals in maxillary first premolars: study of an Andalusian population. AB - A study of 150 extracted maxillary first premolars from citizens of Seville, Andalusia, southern Spain, revealed 60 teeth with one root (40.0%), 85 teeth with two roots (56.7%) and five teeth with three roots (3.3%). The distribution of root canal shapes in the sample showed that all teeth with two or three roots had type I root canals (each canal had one apical foramen). Conversely, most of the single-rooted maxillary first premolars had root canal shape type II (two canals converging in the same apical foramen). Only 1.3% of the teeth had a unique orifice in the pulp chamber and only one root canal. These results emphasized the importance of good knowledge of the root canal morphology and the need for a careful radiographic examination as part of competent root canal therapy of maxillary first premolars. PMID- 10379275 TI - Post-operative symptoms and healing after endodontic treatment of infected teeth using pulsed Nd:YAG laser. AB - Forty-four teeth in 38 patients, diagnosed with chronic apical periodontitis, were endodontically treated. Root canals were shaped using a step-back technique with 5% NaOCl and 3% H2O2 as irrigants. In half of the teeth the canal terminus was irradiated with pulsed Nd:YAG laser (1 W, 15 pps, 1 s). All canals were then obturated with laterally condensed gutta-percha points and sealer, and final radiographs were obtained. Occurrence of spontaneous pain was recorded 1 day after treatment. Percussion pain was recorded after 1 week, and then at 3 and 6 months after treatment. Radiographic follow-up was performed at 3 and 6 months. Percussion pain was significantly less (P < 0.05) in the laser-treated group than in the control group, both 1 week and 3 months after treatment. Other differences between the groups were not significant. These results suggested that the clinical application of pulsed Nd:YAG laser might be advantageous for the treatment of infected root canals. PMID- 10379277 TI - Accuracy of four electronic apex locators: an in vitro evaluation. AB - In the present study, the accuracy and operator dependency of four electronic canal length measuring devices (Apex Finder AFA Model 7005, Apex-Finder, Neosono Ultima EZ and Apit 2) were compared under a set of specified conditions. The electronic apex locators were tested in unflared dry, flared wet and flared dry canals, and in a gelatin as well as in a sodium hypochlorite sponge model. Fifteen extracted single-canaled teeth were selected. The differences between canal lengths obtained by the electronic apex locators and actual canal lengths were scored. Only the Apex-Finder was found to be unreliable (measurements higher than +/- 0.5 mm from the apical foramen). This device was also found to be particularly dependent on operator. A ranking based on a precision of +/- 0.1 mm from the apical foramen showed the Apex Finder AFA Model 7005 to be the most accurate. Early coronal flaring did not ensure better or more precise readings. The gelatin model was evaluated to be more suitable for testing electronic apex locators in vitro than the sodium hypochlorite model. PMID- 10379276 TI - Trauma to primary teeth of South African pre-school children. AB - A clinical survey of 1466 children of typical South African communities showed a prevalence rate of primary tooth trauma of 15%. Trauma was least common at age 1 2 years (10.7%) and most common at 4-5 years (20.6%). The commonest trauma seen was fracture of enamel only (71.8%) followed by fracture of dentine (11.2%), tooth loss (8.2%) and discolouration without other sign of injury (5.6%). PMID- 10379278 TI - Biomechanics of endodontic endosseous implants--a comparative photoelastic evaluation. AB - Dental biomechanics is an interdisciplinary study wherein engineering principles are used for the better understanding of clinical dentistry. The present biomechanical study was done to understand the mechanism by which an endodontic implant transmits occlusal forces to the surrounding bone. In this experimental study, photoelastic techniques were utilized to compare stress distribution patterns in the supporting bone of an intact tooth, a tooth with supporting bone loss, and a tooth stabilized using an endodontic endosseous implant. It was concluded that there were distinct variations in the biomechanics underlying various dental clinical conditions. Further, the implant did not appear to improve the stress distribution. PMID- 10379279 TI - Complex treatment of dens invaginatus type III in maxillary lateral incisor. AB - The complex anatomy and diagnosis of dens invaginatus make endodontic treatment of such teeth difficult. This case describes combined nonsurgical and surgical treatment of a maxillary lateral incisor with a normally shaped canal and a dens invaginatus type III with a lateroradicular lesion. The root canal was treated conventionally with gutta-percha and a zinc oxide-eugenol sealer. The root was surgically exposed and the canal of the dens invaginatus was cleaned, instrumented and obturated with gutta-percha and a zinc oxide-eugenol sealer. At follow-up 3 years 6 months later, the tooth was asymptomatic and radiographically showed repair of the lesion in the region of the dens invaginatus. PMID- 10379280 TI - Severe lateral luxation and root fracture: report of a case with 5-year follow up. AB - A case of severe lateral luxation and root fracture in upper incisors is reported. Treatment involved the repositioning and fixation of the injured teeth and endodontic treatment with calcium hydroxide. The importance of long-term follow-up is emphasized. PMID- 10379281 TI - Pluralism--a collision of ideas. PMID- 10379282 TI - Enamel matrix derivative for periodontal reconstructive surgery: technique and clinical and histologic case report. AB - This paper describes a step-by-step technique for the application of Emdogain, a new enamel matrix derivative (EMD) graft material, for periodontal reconstructive surgery. A case report is presented with a 1-year follow-up. The rationale for use and advantages and disadvantages of EMD are discussed. An additional human histologic case report demonstrates that the formation of new bone, cementum, and periodontal ligament is possible following the use of EMD. PMID- 10379284 TI - A new concept for etching in restorative dentistry? AB - The purpose of the present study was to investigate the scanning electron microscopic texture of enamel and dentin cavity surfaces in extracted human teeth following different etching modalities, specifically combinations of etchants adapted to the tissue composition of the cavity walls. It was concluded that an etching technique that combined the action of 2 different etchants- ethylenediaminetetraacetic acid (EDTA) on dentin and phosphoric acid on enamel- optimized retention structures on each tissue surface of a dental cavity better than either 1 of the 2 etchants that were applied to both types of tissue in the cavity walls. PMID- 10379283 TI - Restoring the gingival contour by means of provisional resin crowns after single implant treatment. AB - A consecutive group of 55 patients was treated with 63 single-implant restorations. The soft tissue was allowed to heal to either provisional resin crowns (n = 25) that were placed at the time of second-stage surgery, or to healing abutments (n = 38) before final crown insertion. An index that assessed the size of the interproximal mucosa adjacent to the single-implant restorations was used to evaluate the volume of the papillae 2 years after crown insertion. The results indicated that the use of provisional crowns may restore soft tissue contour faster than healing abutments alone, but the papillae adjacent to single implant restorations presented similar volume in both groups after 2 years in function. Furthermore, the mean marginal bone loss at the implants was 0.9 mm after 1 year, and no differences were observed between the 2 groups. The present data focus on the need for more scientific data to evaluate different clinical procedures for optimizing esthetic results in implant dentistry. PMID- 10379285 TI - Functional and esthetic outcome enhancement of periodontal surgery by application of plastic surgery principles. AB - The closure of surgical wounds in a layer-by-layer fashion, a common principle of plastic surgery, is applied in this article to the field of periodontal surgery with the introduction of a new flap design. The suggested technique is indicated with all periodontal procedures that aim for hard and soft tissue augmentation (guided bone regeneration, mucogingival surgery, or plastic periodontal surgery) where passive, tension-free wound closure is fundamental for wound healing and a successful functional and esthetic outcome. By means of a series of incisions, buccal and lingual flaps are split several times; this results in a double partial thickness flap and a coronally positioned palatal sliding flap, respectively. Thus, several tissue layers are obtained and the passive advancement of flaps becomes possible for the coverage of augmented areas. Wound closure with microsurgical suture material is accomplished in a multilayer approach, which ensures adaptation and closure of the outer tissue layers without any tension. Two case reports demonstrate the new plastic periodontal approach. PMID- 10379286 TI - Postloading behavior of regenerated tissues in GBR-treated implant sites. AB - The objective of this study was to assess, using reentry procedures, the capacity of regenerated tissues in implant-associated defects to respond to occlusal loading. Two groups of patients treated with membrane-augmented osseointegrated implants were included in the study. In group A (7 patients), a total of 9 implant-associated defects, including 6 dehiscences and 3 immediate extraction sites, were prospectively followed up 6 months following prosthesis connection. In group B (3 patients), 4 dehiscence defects were retrospectively evaluated 5 years after prosthetic loading. All defects in both groups had an uneventful healing period beyond the 6 months following implant insertion and showed complete fill with bone-like hard tissues at abutment connection surgery. A second surgical reentry was carried out to evaluate the quantitative changes in the regenerated tissues at the membrane-treated sites; it was carried out 6 months following prosthesis connection in group A, and 5 years postloading in group B. At the second reentry procedure, the mean percentage of defect fill at the dehiscence sites was 82% +/- 12.8% in group A and 83% +/- 7.3% in group B. In the 3 immediate extraction sites in group A, the most apical bone-implant contact around the implant was consistently located at about 1 mm, relative to the coronal aspect of the implant shoulder, as evidenced both radiographically and during the second reentry. The trends noted in this investigation suggest that tissues regenerated in successfully treated implant-associated defects can be maintained in the short-term and long-term periods following prosthetic loading. PMID- 10379287 TI - The use of biodegradable polylactic acid barrier materials in the treatment of grade II periodontal furcation defects in humans--Part II: A multicenter investigative surgical study. AB - This study evaluated whether differences in design of 3-dimensional polylactic acid barriers (EPi-Guide and Guidor) would influence hard tissue results in the treatment of Grade II furcations in humans. A multicenter study was conducted, using 40 patients with moderate to advanced bilateral chronic adult periodontitis of the mandibular first or second molars. After flap access, debridement, and root preparation, surgical bone level measurements were taken and membranes were placed on a random basis. Surgical reentry occurred at 1 year. Data collected from all 3 investigative centers were pooled and analyzed using an analysis of variance appropriate for a counterbalancing design. Both barrier materials resulted in significant gains of attachment level and defect reduction. The composite reduction in the vertical component of the osseous defects was greater in the sites treated with Epi-Guide as compared to those treated with Guidor; the difference between barriers reached statistical significance (P = 0.02). PMID- 10379288 TI - Case reports offer a challenge to treatment strategies for immediate implants. AB - The placement of osseointegrated implants in extraction sockets is a commonly used and reliable procedure. Many operative protocols have been suggested for use with both submerged and nonsubmerged implants, and some prerequisites have been defined for their successful placement. Dealing exclusively with implants placed in intact extraction sockets, this paper reviews these commonly suggested prerequisites, discusses their clinical relevance, and presents case reports in which clinical success was obtained despite the violation of more than 1 of these factors. Techniques to obtain primary implant stability, procedures to regenerate residual bone defects, the need to submerge implants in the healing phase, and treatment strategy in infected sites are reviewed. Because the simultaneous violation of some prerequisite factors allows postextractive implants to be performed with a single surgical approach, a new classification is proposed based on the number of surgical stages required to replace a failing tooth with an implant-supported restoration. PMID- 10379289 TI - Root trunk dimensions of 5 different tooth types. AB - The primary purpose of this study was to document mean, standard deviation, and range of root trunk dimensions of multirooted tooth types. A total of 412 extracted teeth were examined and classified as: maxillary first molars, maxillary second molars, maxillary first premolars, mandibular first molars, and mandibular second molars. The distance from the cementoenamel junction (CEJ) to the root groove and from the CEJ to the root division was measured. Mean CEJ to root groove distances ranged from 1.35 to 1.65 mm for maxillary first molars, from 1.49 to 1.89 mm for maxillary second molars, from 1.71 to 1.73 mm for maxillary first premolars, from 1.16 to 1.22 mm for mandibular first molars, and from 1.53 to 1.76 mm for mandibular second molars. PMID- 10379290 TI - Esthetic restoration of a single-tooth dental implant using a subepithelial connective tissue graft: a case report with 3-year follow-up. AB - This case report describes the use of a subepithelial connective tissue graft to restore the gingival papillae and augment ridge soft tissues adjacent to a dental implant. The patient was referred for periodontic and prosthodontic evaluation after the placement of an implant--with the implant head 6.5 mm below the adjacent cementoenamel junction--in an area of inadequate bone volume with deficient interproximal papillae and ridge soft tissues. The resulting esthetic defect was restored by means of a combined technique that used a subepithelial connective tissue graft and an emergence profile-contoured crown. A 3-year clinical follow-up with complete regeneration of the gingival papillae is described. PMID- 10379291 TI - Aesthetic soft tissue integration and optimized emergence profile: provisionalization and customized impression coping. AB - In the anterior region, the successful replacement of a single tooth with an implant-supported restoration is one of the most difficult treatment options due to numerous functional and biological requisites. Such prostheses should also satisfy the increasing aesthetic demands of patients who expect the definitive restoration to mimic the natural dentition and their supporting gingival tissues. This article highlights the importance of the provisional restoration as the critical element for aesthetic success and gingival integration. Its perfect replication in the definitive restoration requires the use of optimized transference techniques. PMID- 10379292 TI - Single-tooth implant in the anterior region for optimal aesthetics. PMID- 10379294 TI - Aerosol and vapor control in the dental treatment room. AB - As dental clinicians, common sense is still the best weapon for infection control -the use of antimicrobial cleaning products remains an integral component of any infection control procedure. PMID- 10379293 TI - Restoration of the anterior maxilla with ultraconservative veneers: clinical and laboratory considerations. AB - One significant challenge in aesthetic dentistry is to integrate individual restorations with the adjacent natural dentition. In order to achieve a seamless result postoperatively, the clinical preparation and the laboratory fabrication phases of treatment must be performed in concert. The use of ultraconservative laminate veneers is a restorative modality that permits the development of a functional, aesthetic outcome while preserving the greatest degree of natural tooth structure. This article highlights the preparation design and material considerations that are involved with ultrathin porcelain laminate veneer restorations. PMID- 10379295 TI - Dental implant placement based on restorative components. PMID- 10379296 TI - Direct reconstruction of the maxillary anterior dentition with composite resin: a case report. AB - Direct composite restorations continue to be a viable treatment alternative for numerous patients who desire anterior reconstructive procedures. In order to achieve optimal aesthetic and functional results, the clinician must possess a thorough understanding of composite resins and color and be able to simulate the optical properties of a natural tooth. This paper describes a sophisticated polychromatic color layering technique through the correct implementation of composite materials and freehand bonding techniques for the fabrication of ten direct resin veneer restorations in the maxilla. PMID- 10379297 TI - Posterior Class II composite restorations utilizing a custom occlusal matrix. PMID- 10379298 TI - In vitro study of the retention and mechanical fatigue behavior of four implant overdenture stud-type attachments. AB - This study sought to determine the influence of mechanical fatigue on four varieties of implant overdenture studtype attachments (Supra-Snap, O'Ring, TSIB, ZAAG). Measurements of the initial vertical retentive force and the weight of the implant abutment were recorded. The same procedure was performed after the equivalent of 2 months, 6 months, and 12 months of clinical wear. For the four attachments, weight variation of the abutment between 0 and 1,080 cycles demonstrated no significant difference. Results indicated the TSIB to be significantly most retentive; next most retentive was the O'Ring, followed respectively by Supra-Snap and ZAAG. PMID- 10379299 TI - The intertrochanteric osteotomy and pseudarthrosis of the femoral neck. 1957. PMID- 10379300 TI - Bernese periacetabular osteotomy. AB - Seventy-five symptomatic dysplastic hip joints (63 patients) were treated with the Bernese periacetabular osteotomy during a period of 44 months. The mean patients' age was 29 years (range, 13-56 years) and the female:male ratio was 3.4:1. Group III dysplasia according to Severin was seen in 50% and Group IV dysplasia was seen in 44% of the patients. Osteoarthritis was present in 58% of the patients. Followup was obtained at a mean of 11.3 years (range, 10-13.8 years) in 71 hip joints (95%). Radiographic measurements of the lateral center edge angle, anterior center edge angle, acetabular index, lateralization of the femoral head, and intactness of Shenton's line showed a high correction potential of this type of osteotomy. In 58 patients (82%) the hip joint was preserved at last followup with a good to excellent result in 73%. Unfavorable outcome was significantly associated with higher age of the patient, moderate to severe osteoarthritis at surgery, a labral lesion, less anterior coverage correction, and a suboptimal acetabular index. Major complications were encountered in the first 18 patients including an intraarticular cut in two, excessive lateralization in one, secondary loss of correction in two and femoral head subluxation in three patients. PMID- 10379301 TI - Periacetabular osteotomy through the Smith-Petersen approach. AB - Sixty-six hips in 58 patients that had undergone periacetabular osteotomy for residual acetabular dysplasia were available for clinical and radiographic followup at a minimum of 2 years (average, 4 years) after surgery. The final clinical results were graded as 17% excellent, 59% good, 12% fair, and 12% poor. No patient who met the ideal indications for surgery had a poor result during the study period. After reviewing the results, the authors remain positive regarding the periacetabular osteotomy and recommend it for individuals with hip pain and radiographic evidence of acetabular dysplasia. PMID- 10379302 TI - Complications of periacetabular osteotomy. AB - There was a statistically significant decrease in major complications from 17% to 2.9% when comparing the first 35 cases with the second 35 cases of periacetabular osteotomy performed by one surgeon. There were no cases of intraarticular fracture, conversion to total hip replacement, or deaths in this series. Of considerable significance was that almost all major complications, as defined for disclosure in this report, left the patients with no permanent sequelae after either successful treatment, as in intraoperative bleeding, or with observation with time, as for recovery of sciatic nerve function. The complication rate of periacetabular osteotomy decreases significantly in proportion to increasing experience, as documented in this study. Patients in ongoing studies completed the Western Ontario and McMaster Universities Osteoarthritis Index and the Short Form- 36 preoperatively, which will add to the authors' ability to comment on functional outcomes in future reports. PMID- 10379303 TI - Surgical correction of acetabular dysplasia in the adult. A Boston experience. AB - Acetabular redirection surgery is the mainstay of treatment for the symptomatic, dysplastic hip. The authors' experience with the Salter innominate osteotomy, Wagner spherical acetabular osteotomy and the modified Bernese periacetabular osteotomy shows that major acetabular redirection surgery can reliably improve the structure of the dysplastic hip and delay or prevent secondary osteoarthrosis. The limited correction achieved by the Salter innominate osteotomy suggests this procedure generally should be reserved for younger patients with mild dysplasia. The modified Bernese periacetabular osteotomy is the authors' current preferred method of treating acetabular dysplasia, even in the presence of mild to moderate secondary osteoarthrosis. PMID- 10379304 TI - Early experience and results with the periacetabular osteotomy. The Mayo Clinic experience. AB - The purpose of the present study was to review the early results of periacetabular osteotomy in the initial group of patients undergoing this procedure at the authors' institution. The first 21 hips in 19 patients with greater than 2 years followup, which represents the learning curve with this operation, were reviewed retrospectively. There were 14 females and five males with an average age of 21 years (range, 17-43 years). Intertrochanteric osteotomy was performed simultaneously on four patients with coxa valga and inadequate correction with periacetabular osteotomy alone. At an average of 38 months of followup (range, 24-52 months), the Mayo hip scores improved from an average of 46 points (range, 34-58 points) to an average of 68 points (range, 42-80 points). Hip range of motion declined slightly in all three arcs of motion. The lateral center edge angle of Wiberg improved from an average of 2 degrees to an average of 24 degrees. The loading zone angle (Tonnis) improved from an average of 24 degrees to an average of 11 degrees. The anterior center edge angle of Lequesne improved from an average of -6 degrees to an average of 38 degrees. Complications included two peroneal palsies, both of which resolved completely; three ischial fractures that healed uneventfully; three asymptomatic pubic nonunions; and asymptomatic heterotopic ossification in five patients. One patient underwent subsequent total hip arthroplasty for progressive arthritis and pain. Another patient required intertrochanteric osteotomy at a later date. The early results in this initial group of patients treated with periacetabular osteotomy show reliable radiographic correction of deformity and improved function with an acceptable complication rate. Patients should be counseled carefully about possible loss of motion postoperatively. Additional study is necessary to assess the long term results of this procedure. PMID- 10379305 TI - The periacetabular osteotomy. Minimum 2 year followup in more than 100 hips. AB - The results of 123 periacetabular osteotomies in 115 patients were reviewed at an average clinical followup of 4.3 years. The average age of the patients at the time of the operation was 32.9 years. The preoperative diagnosis was congenital dysplasia in 101 hips, Legg-Calve-Perthes disease in 10 hips, Charcot Marie Tooth disease in four hips, epiphyseal dysplasia in three hips, congenital coxa vara in two hips, slipped capital femoral epiphysis in one hip, and posttraumatic and postinfectious dysplasia in one hip each. The ilioinguinal approach was used in 67 hips and the modified Smith-Petersen approach was used in 56 hips. A periacetabular osteotomy was combined with an intertrochanteric osteotomy and/or trochanteric transfer in 32 hips. Ten hips underwent open reduction and internal fixation of an acetabular rim fracture and 18 arthrotomies were performed at the time of periacetabular osteotomy. The average Harris hip score increased from 65 points preoperatively to 89 points at latest followup. The average Merle d'Aubigne score increased from 13.6 points preoperatively to 16.3 points at latest followup. Overall, 83% of the hips were rated clinically as good to excellent. Seven hips have undergone total hip arthroplasty and six subsequent intertrochanteric osteotomies were performed. The majority of the major complications occurred when the osteotomy was performed through the ilioinguinal approach. The latest followup radiographic severity of osteoarthrosis, according to the criteria of Tonnis, improved or was unchanged in 117 hips (95%), and progressed in only six hips (5%). The majority of the hips with preoperative changes in the periarticular bone showed some evidence of regeneration, which was shown by a decrease in the subchondral sclerosis, disappearance of cysts, or healing of an acetabular rim fracture. The short term results of the periacetabular osteotomy are encouraging from the standpoint of improvements in clinical scores and in the appearance of the joint. PMID- 10379306 TI - A comparison of different surgical approaches for the periacetabular osteotomy. AB - The periacetabular osteotomy is a well established surgical procedure for the preventative treatment of degenerative joint disease caused by symptomatic acetabular dysplasia. Surgeons on several continents use varying surgical approaches to achieve the same effective osteotomy. Individual surgical approaches must provide accurate and adequate exposure for the osteotomy and the reorientation of the acetabular fragment. The aim of the surgical approach for such complex and expansive surgery is to minimize morbidity related to the approach. This article compares experiences among three common approaches including the modified Smith-Petersen, ilioinguinal, and direct anterior approaches and describes the double approach. PMID- 10379307 TI - Results of periacetabular osteotomy in patients with previous surgery for hip dysplasia. AB - The results of 19 periacetabular osteotomies in 18 patients who had undergone prior bony surgical procedures for hip dysplasia were evaluated. There were 10 females and eight males with an average age of 30.9 years. Previous surgical interventions included 18 intertrochanteric osteotomies, nine pelvic osteotomies, and two shelf acetabuloplasties. The average clinical followup for this group was 45 months. Harris hip score averages improved from 60 to 90 points. Merle d'Aubigne scores showed similar elevations from 13.1 to 16.4 points. Radiographic assessment documented increased coverage and lower Tonnis secondary arthrosis grades in a significant number of hips. No significant differences in outcome were found between this group and a reference group of patients undergoing periacetabular osteotomy who had no previous hip surgery. These intermediate term results are encouraging and seem to discount anticipated problems of prior scarring and distorted pelvic and proximal femoral anatomy. PMID- 10379308 TI - Technical complications of the Bernese periacetabular osteotomy. AB - Periacetabular osteotomy provides technical challenges in surgical exposure and reorientation of the acetabular fragment. There is evidence to indicate that the procedure is effective, but there is a recognized learning curve with respect to its performance. This analysis includes a retrospective review of the technical complications encountered in more than 500 cases. These problems were analyzed and then related to the stages of the procedure in which they occurred (surgical approach, osteotomy, fragment positioning, fragment fixation, and the postoperative period). Efforts are made to analyze the cause of the complications and to suggest preventative or remedial treatments. It is hoped that highlighting these complications will shorten the learning curve for surgeons trying this procedure for the first time. PMID- 10379309 TI - Anterior femoroacetabular impingement after periacetabular osteotomy. AB - As experience with the Bernese periacetabular osteotomy has grown, an unexpected observation in a group of patients has alerted the authors to the risk of a secondary impingement syndrome that may occur some time after the periacetabular osteotomy. This possibly may explain residual pain and limited range of motion in a larger group of patients. The impingement is produced by abutment of the femoral head or head to neck junction on the anterior rim of the properly aligned acetabulum. The symptoms are those of restricted flexion, and limited or absent internal rotation in flexion, with variable groin pain. Magnetic resonance imaging studies may reveal acetabular labral disease and adjacent cartilage damage associated with the impingement. Lack of anterior or anterolateral offset between the femoral neck and head results in neck to rim contact when the hip is flexed and/or internally rotated. Before the periacetabular osteotomy this is compensated by the lack of anterior acetabular coverage, but after proper correction the mismatch becomes apparent. The authors recently have devised a routine during the periacetabular osteotomy procedure whereby after the acetabular fragment is corrected into the desired position, the joint is opened, visually inspected, and palpated for impingement with the hip flexed and internally rotated. When necessary, a resection osteoplasty of the femoral neck to head junction is performed to improve the head and neck offset and reduce the anterior contact. This, in the short term, has provided satisfactory prevention of postoperative impingement. PMID- 10379310 TI - Distal biceps ruptures. A followup of Boyd and Anderson repair. AB - Twenty-one distal biceps ruptures in 20 patients were treated during a 10-year period. All patients were men with an average age of 47 years (range, 37-70 years). Long term results (average, 44 months) were assessed using the American Academy of Orthopaedic Surgeons Disabilities of the Arm, Shoulder, and Hand outcomes questionnaire, isokinetic testing of elbow flexion and supination (strength and endurance), and patient satisfaction. Measurements of range of motion revealed diminished forearm rotation in four of 21 (19%) elbows and diminished flexion in one of 21 (5%). Isokinetic testing revealed 10 of 21 (48%) elbows had weakness of supination versus three of 21 (14%) for flexion. Deficits in endurance included eight of 21 (38%) elbows for supination and seven of 21 (33%) for flexion. Seven complications occurred in seven patients for a 35% complication rate. Heterotopic ossification was the most common complication, occurring in three patients. One of these patients also had a synostosis. All patients completed the Disabilities of the Arm, Shoulder, and Hand outcomes questionnaire. Twelve patients had an excellent outcome, eight had a good outcome, and no patient had a fair or poor outcome. These results correlated closely with the patients' own subjective assessments of their satisfaction with the procedure. Despite diminished strength and endurance and a high rate of complications, patient satisfaction was excellent and functional outcome was good. PMID- 10379311 TI - Long term outcome of osteochondritis dissecans of the humeral capitellum. AB - Long term outcome of osteochondritis dissecans of the humeral capitellum was determined for 53 patients. The average age of the patients at the time of treatment was 16.6 years (range, 10-34 years). The average followup was 12.6 years (range, 3-25 years). Seven of 14 (50%) patients who were treated conservatively and 18 of 39 (46%) who were treated by surgical removal of the fragment were found to have residual elbow symptoms associated with daily living activities (poor outcome). The initial radiographs of the elbow were available for 45 patients; a poor outcome was seen in six of 19 (32%) early lesions and 13 of 26 (50%) advanced lesions. Fourteen elbows had evidence of osteoarthritis on the initial radiographs, and a poor outcome was seen for nine (64%) of these elbows. After removal or detachment of the fragment, seven osteochondral defects were assessed as large, and all seven had a poor outcome. These long term results suggest that the residual elbow symptoms associated with daily living activities in approximately 50% of patients may be associated with advanced lesions, osteoarthritis of the elbow, and a large osteochondral defect. PMID- 10379312 TI - Volar dislocation of the metacarpophalangeal joint of the ring finger. Report of two cases. AB - Volar dislocation of the metacarpophalangeal joint of the finger is a rare injury. Two cases of an isolated volar dislocation of the ring finger metacarpophalangeal joint are reported. The dislocation was treated successfully by closed reduction shortly after injury in both cases. Closed reduction should be attempted in all cases of this injury because successful reduction may be possible. PMID- 10379313 TI - Repeat chemonucleolysis is safe and effective. AB - A review was made of 85 patients who received a second injection of chymopapain because of a recurrent disc herniation between 1980 and 1996. All patients were pretreated for 3 days with H1 and H2 receptor blockers. Immediate sensitivity reactions were not seen. Four Type 1 and one Type 2 reactions were seen after the the second injection. No other complications were seen. In this group of 85 patients, 66 patients received a second injection at the same level as primary treatment after a period of 57.1 months (range, 2-143 months). Two patients were lost to followup. Four patients had surgery, three with good results. Good results and no complications were seen in three patients who had a third chemonucleolysis because of another recurrence after 15, 40, and 56 months, respectively. The remaining 57 patients were interviewed after 64 months (range, 3-143 months). Using the Prolo scale, 51 patients were rated as having excellent or good results, and six patients were rated as having fair or poor results. PMID- 10379314 TI - Severe hyperflexion sprains of the lower cervical spine in adults. AB - Severe sprains of the cervical spine result from a traumatic rupture of the intervertebral disc and ligaments. Although rare, these lesions may lead to a significant kyphotic deformity if they are not surgically treated. The treatment of such a kyphotic deformity may consist of surgical fixation of the lesion through either an anterior or posterior approach. A retrospective study has been done examining 44 severe cervical sprains in 41 patients surgically treated through a posterior approach, using Roy-Camille plates. With an average followup of 29 months (range, 6-60 months), 73% of the patients recovered a normal range of spinal motion, with either moderate or no pain. No neurologic or vascular complications directly attributable to posterior plating and no secondary kyphosis were observed. A moderate sagittal displacement with kyphotic angulation occurred above the fusion in five patients. Posterior screw plate fixation appears to be a safe and effective treatment for severe hyperflexion sprain of the lower cervical spine in the adult. PMID- 10379315 TI - Relationship of acetabular wear to osteolysis and loosening in total hip arthroplasty. AB - Polyethylene wear and the subsequent development of periprosthetic osteolysis are the major factors limiting the longevity of total hip arthroplasties. A minority eventually loosen, but no clinically applicable system exists for accurate early prediction of failure. The relationship between acetabular wear and the development of loosening, osteolysis, and revision was investigated in 235 Charnley low friction arthroplasties. The average age of the patient at surgery was 31.7 years (range, 17-39 years), and the duration of followup averaged 234 months (19.5 years; range, 74-364 months). Total wear averaged 2.1 mm (range, 0 7.2 mm), and the average wear rate was 0.11 mm per year (range, 0-0.55 mm/year), with the wear rate of revised components being twice that of surviving ones (0.19 mm/year versus 0.09 mm/year). The prevalence of osteolysis (33 hips, 14%) and of acetabular and femoral component loosening and revision rose significantly with increasing wear. Osteolysis also was associated significantly with femoral component loosening and revision, but the presence of calcar changes was not (90 hips, 38%). Twenty-five year survivorship exceeded 90% for arthroplasties with a wear rate less than 0.1 mm per year, but 20-year survivorship of acetabular components with a rate greater than 0.2 mm per year was below 30%, and none survived 25 years. For every additional millimeter of wear, the risk of acetabular revision in any one year increased by 45% and for the femur increased by 32%. PMID- 10379316 TI - Greater trochanteric blood flow during total hip arthroplasty using a posterior approach. AB - The authors investigated the effect of a posterior surgical approach on the local femoral blood supply during primary total hip arthroplasty. Greater trochanteric blood flow measurements were made with a laser Doppler flowmeter at intervals during the performance of eight uncemented and nine cemented total hip arthroplasties. Complete detachment of the quadratus femoris was associated with a significant decrease in trochanteric blood flow in the uncemented and cemented groups. The lowest perfusion levels during the procedure were seen transiently with posterior dislocation of the femoral head, after which trochanteric perfusion was decreased by 66% in the uncemented group, and 61% in the cemented group compared with baseline values. Blood flow remained approximately half of baseline values after insertion of the femoral prosthesis in the uncemented and cemented groups. These changes in greater trochanteric blood flow may serve as a marker for reduction in proximal femoral blood flow during total hip arthroplasty, and subsequently relate to the extent of bony ingrowth, periprosthetic bone loss, and ultimately the incidence of implant failure caused by aseptic loosening. PMID- 10379317 TI - Localization of hyaluronan in pseudocapsule from total hip arthroplasty. AB - Hyaluronan is a large glycosaminoglycan present in normal synovial fluid imparting important viscoelastic properties for joint lubrication. In normal and noninflammatory conditions, hyaluronan has been localized to the lining layer of the synovial membrane, specifically synovial fibroblasts. Prosthetic joint fluid contains large amounts of hyaluronan, the source of which has not been determined. Pseudocapsules from patients who had total hip arthroplasty and were having revision surgery for nonloosened and nonseptic conditions were fixed in 10% acid formalin with 70% alcohol and stained for hyaluronic acid with biotinylated hyaluronic acid binding protein as a probe. Hyaluronan was localized strongly to the lining layer of the pseudocapsule, as in normal and osteoarthritic synovial membranes. The fibroblasts in the pseudocapsule of total hip arthroplasty may be responsible for the persistent production of hyaluronan in prosthetic joint fluid. PMID- 10379318 TI - Omnifit-HA stem in total hip arthroplasty. A 2- to 5-year followup. AB - Outcomes of the first 60 noncemented Omnifit-HA total hip arthroplasties in 56 patients were studied prospectively for 2 to 5 years. The femoral prosthesis had a proximal third circumferential hydroxyapatite coated surface treatment. The acetabular component was a hemispheric modular, porous, nonhydroxyapatite press fit cup, supplemented with screw fixation. One cup was revised for recurrent dislocation, with no femoral revisions. The mean Harris hip score was 54 (range, 20-76) before surgery and 96 (range, 83-100) at final followup, with all patients having an excellent or good outcome. Mild thigh pain occurred in 6% of hips. Subsidence occurred in 9% of hips (range, 1-2.8 mm); in all cases, subsidence was nonprogressive after 1 year. Stable bone ingrowth fixation was evident at the hydroxyapatite coated portion in 100%. A sclerotic reactive line adjacent to the nonhydroxyapatite portion of the stem occurred in 81% but was not adjacent to the hydroxyapatite coated portion of any stem. Endosteal condensation occurred in 90% and correlated with a higher Harris hip score (mean score, 96 with, 91 without). Endosteal lysis adjacent to or distal to the hydroxyapatite coating did not occur. Lytic lesions at the calcar occurred in 19% and correlated with a greater linear acetabular polyethylene wear rate (mean, 0.30 mm/year with lytic lesions, 0.17 mm/year without). This noncemented stem with proximal third hydroxyapatite coating showed excellent short term clinical and radiographic outcome. Absence of distal endosteal lysis, along with correlation of calcar erosion to polyethylene wear, suggests that early circumferential bony ingrowth afforded by hydroxyapatite coating prevents distal endosteal access to polyethylene debris at short term followup. PMID- 10379319 TI - Evaluating marrow margins for resection of osteosarcoma. A modern approach. AB - Intraoperative evaluation of bone marrow margins by frozen section analysis is a common practice in the surgical treatment of osteogenic sarcoma. The purpose of this study was to assess the clinical use of intraoperative marrow margin evaluation to rule out occult intramedullary tumor extension in osteosarcoma surgery. One hundred twenty-eight consecutive patients with high grade osteosarcoma diagnosed between 1988 and 1996 (Group 1) were reviewed retrospectively and compared with 92 consecutive patients treated from 1979 to 1984 (Group 2). Eighty-five patients in Group 1 met the inclusion criteria of having high grade intramedullary lesions of the long bones observed on preoperative magnetic resonance imaging evaluation of the lesion and intraoperative frozen section analysis of the bone marrow margin. Thirty-three patients in Group 2 met the same inclusion criteria with the exception of having preoperative magnetic resonance imaging. Ninety-two marrow margins in Group 1 and 33 marrow margins in Group 2 were evaluated by frozen section. All 92 marrow margins in patients in Group 1 were negative by frozen section analysis and permanent histologic analysis. Of the 33 marrow margins in patients in Group 2, three (9.1%) were reported positive for tumor. Of these, one was found to be a false positive result on permanent pathologic examination. In addition, one false negative frozen section result was found, which was positive for tumor on permanent pathologic examination. The difference in true positive results of marrow margins between Group 1 and Group 2 was statistically significant. Intraoperative marrow margin evaluation by frozen section is not mandatory with modern imaging techniques. Preoperative evaluation of tumor extent using magnetic resonance imaging and intraoperative evaluation of the specimen by the pathologist (done by bivalving the specimen) are reliable methods to ensure adequate surgical margins in most cases of conventional osteosarcoma of the long bones. PMID- 10379320 TI - Aneurysmal bone cyst. A population based epidemiologic study and literature review. AB - Aneurysmal bone cyst is a rare nonneoplastic expansile osteolytic bone lesion of unknown etiology. To the best of the authors' knowledge, no epidemiologic study concerning its incidence has been reported. The authors performed a retrospective, population based analysis of 94 patients with primary aneurysmal bone cyst and a literature review of 1002 patients regarding gender and age predilection. The annual incidence of primary aneurysmal bone cyst was 0.14 per 10(5) individuals. The male to female ratio was 1:1.04, and the median age was 13 years (range, 1-59 years). The results of this study and data compiled from the literature show that aneurysmal bone cysts occur significantly more often in female patients. PMID- 10379321 TI - Extracortical bridging callus after limb salvage surgery about the knee. AB - The increasing survival of children treated for osteosarcoma has led to an increase in limb sparing surgery. Little published information is available about the postoperative imaging appearance of this technique. Thus, information gleaned from medical records and imaging reviews was correlated with clinical outcome of 19 consecutive children (median age at diagnosis, 12.3 years) treated for distal femoral osteosarcoma with a cemented rotating hinged knee endoprosthesis with porous coated collar. An extracortical bridging bony callus was identified in 16 patients who were followed up after surgery for a median of 2.8 years (range, 1.4 6 years). Extracortical bridging bony callus formation was circumferential in eight and preferentially posterior in nine, and ranged from 0.5 to 0.7 cm in thickness; 14 patients had lucent lines subjacent to the bridging bone. Thirteen patients had good functional use of the surgically treated leg. Five patients had progressive metastatic disease develop. One patient had local disease recurrence 8 months after surgery. Extracortical bridging bone was not identified in three patients, two of whom had infection develop 7 months and 17 months after surgery; both infections required amputation. Extracortical bridging bone preferentially develops posteriorly, along the compression side of the femur, often is associated with subjacent lucency, and seems to reflect increased prosthetic stability. PMID- 10379322 TI - Fracture reduction and deformity correction with the hexapod Ilizarov fixator. AB - A configuration for the Ilizarov external fixator with six distractors and 12 ball joints in the form of a hexapod was developed. The system allows for six degrees of freedom bone fragment displacement by controlling the distractors. Using this assembly, universal three-dimensional corrections or reductions are possible without the need for complicated joint mechanisms. The device was used in 16 patients: five had displaced tibial fractures with severe soft tissue damage, 10 had deformities or pseudarthroses subsequent to treatment of tibial fractures, and one had an axis deviation in the course of tibial lengthening. Translational (to 40 mm) and rotational deformities (to 33 degrees) were corrected. Final radiographic examinations after the correction procedure was complete showed median residual deformities of 3.5 mm (range, 0-5 mm) and 1 degree (range, 0 degree-4 degrees) in the anteroposterior projection and of 1.5 mm (range, 0-6 mm) and less than 1 degree (range, 0 degree-9 degrees) in the lateral projection. The construction is a useful and important addition to the Ilizarov fixator system. As a bone fixation device it is unique in that its optimal use depends on the availability of computer software. PMID- 10379323 TI - Haemophilus aphrophilus osteomyelitis after dental prophylaxis. A case report. AB - A 36-year-old patient who was otherwise healthy had acute osteomyelitis of the humeral shaft develop after routine prophylactic dental cleaning and ultrasonic scaling. Haemophilus aphrophilus grew on cultures of material obtained during biopsy of the humerus, and pathologic examination confirmed the diagnosis of acute osteomyelitis. Haemophilus aphrophilus, a fastidious gram negative bacillus, is part of the normal oral flora and is a rare pathogen. Osteomyelitis caused by Haemophilus aphrophilus has not been reported to occur after routine dental prophylaxis. The patient was treated successfully with surgical debridement and appropriate antibiotics. PMID- 10379324 TI - Obturator neuropathy. An anatomic perspective. AB - Entrapment of the anterior division of the obturator nerve is a recently described cause of medial groin pain. This anatomic study examines the extrapelvic course of the nerve and related fascia in the adductor region to provide an anatomic basis for the syndrome and to aid in surgical treatment. Twelve anatomic specimen limbs were dissected to document the extrapelvic course of the obturator nerve, the myofascial arrangement, and the vasculature. A thirteenth limb was prepared with intraarterial glycerin to examine the vessels in more detail. A distinct fascial plane was found deep to the adductor longus and pectineus overlying the anterior division of the obturator nerve. The arterial supply to the adductor muscles is related intimately to the nerve and its branches, with associated local thickening of the fascial connective tissue. The relationship between the nerve, vessels, and fascia appears sufficient to result in an entrapment syndrome. The anatomic findings from this series will help plan the surgical treatment of this condition. PMID- 10379325 TI - The vascularization of the os calcaneum and the clinical consequences. AB - This study was conducted to analyze extraosseous and intraosseous vascularization of the os calcaneum and to elucidate possible clinical manifestations. The arteries of 13 lower leg and foot specimens of human cadavers were injected with a polymer and subjected to maceration or were embedded in plastic. The examination revealed that 45% of the bone is vascularized via medial arteries and 45% via lateral arteries, whereas the remaining 10% is supplied by the sinus tarsi artery. From the medial side, two or three vessels branch off the posterior tibial artery, penetrate the calcaneus below the sustentaculum, and supply the medial part of the posterior joint. The lateral calcaneal artery normally is a branch from the posterior tibial artery. In two of 13 specimens, this lateral supply comes from the peroneal artery. The medial and lateral intraosseous arterial supply for the calcaneus is equal. Inside the bone there is a water-shed zone where the medial and lateral arterial supply meet in the midline. Only 10% of the blood flow is supplied by vessels in the sinus tarsi. Clinically, interruption of the lateral calcaneal artery during the conventional lateral surgical approach for a calcaneus fracture may result in ischemic bone necrosis. The lateral calcaneal artery could supply a local microvascular flap to cover soft tissue defects of the heel. A compartment syndrome after a calcaneus fracture may be caused by bleeding from the medial calcaneal arteries into the quadratus plantae compartment. PMID- 10379326 TI - Incidence and mechanism of the pivot shift. An in vitro study. AB - The aim of this study was to determine the incidence and mechanism of the pivot shift phenomenon in the normal and anterior cruciate ligament transected knee in vitro. Fifteen knees were tested under a range of valgus moments and iliotibial tract tensions when intact and after anterior cruciate ligament transection. Knee kinematics were measured and described in terms of tibial rotation as the knee flexed. Eight knees pivoted after anterior cruciate ligament transection. The mean pivot shift motion was an external tibial rotation of 17 degrees (+/- 11 degrees standard deviation) over a range of 27 degrees (+/- 24 degrees) knee flexion, at a mean flexion angle of 56 degrees (+/- 27 degrees). Clinically, this corresponds to a reduction of an anteriorly subluxed lateral tibial plateau as the knee flexes. When intact, pivoting and nonpivoting knees had similar anteroposterior laxity, but after anterior cruciate ligament transection, the pivoting group had significantly greater laxity. The loading required to elicit the pivot shift was critical and variable between knees, which raises questions about comparing clinicians' techniques and results in assessing the buckling instability attributable to anterior cruciate ligament injury. PMID- 10379327 TI - Prolonged leaching time of peptide antibiotics from acrylic bone cement. AB - The leaching time of antibiotics in growth inhibitory concentrations from polyacrylic bone cements was determined using Escherichia coli, methicillin resistant Staphylococcus aureus, Staphylococcus epidermidis, Klebsiella pneumoniae, and Pseudomonas aeruginosa as target bacteria. The leaching time of the peptide antibiotics vancomycin and polymyxin B nonapeptide was considerably longer than that of gentamicin, novobiocin, and erythromycin. Among the nonpeptide antibiotics, the leaching time of novobiocin lasted longer than did that of gentamicin. The acylated parent polymyxin B antibiotic leached for a considerably shorter period than did the deacylated polymyxin B nonapeptide derivative, suggesting that the lipids impede the leaching process from the cement. Cement beads impregnated with polymyxin B nonapeptide and introduced into the tibia of three rabbits receiving oral novobiocin protected bone infection against a challenge of Pseudomonas aeruginosa that otherwise caused infection in tibias containing gentamicin impregnated cement beads. The peptide antibiotics alone or in combination with other antibiotics (polymyxin B nonapeptide and novobiocin) impregnated in cements for orthopaedic procedures may provide longer periods of protection against a wide range of bacterial pathogens. PMID- 10379328 TI - Biomechanical analysis of prophylactic fixation for middle third humeral impending pathologic fractures. AB - For determination of the most biomechanically desirable construction for prophylactic fixation of impending central 1/3 humeral fractures, 24 matched pairs of fresh frozen skeletonized human cadaveric humeri were divided randomly into four groups. Group 1 compared intact humeri with matched humeri that had a 50% hemicylindrical cortical central 1/3 defect to show reproducible failure at the defect with significant reduction in strength. Groups 2 through 4 compared prophylactic fixation of the defect combined with cementation and dynamic compression plating, Rush rodding, or locked intramedullary nailing. Each specimen was tested in external rotation torsion to failure by fracture. In Group 1, test specimens with defects failed with significantly lower rotation to failure, peak torque, stiffness, and total energy absorbed to failure. In Groups 2 through 4, intramedullary nailing provided statistically significantly better total energy absorbed to failure and stiffness than did dynamic compression plating. The proximally and distally locked intramedullary nail seems to have biomechanical advantages in the prophylactic stabilization of an impending pathologic fracture of the central 1/3 of the humerus. These biomechanical findings must be considered in light of the clinical context when a means of fixation is selected. PMID- 10379329 TI - Evaluation of tibial component fixation in specimens retrieved at autopsy. AB - Fixation of the tibial component was evaluated in eight knees retrieved at autopsy by comparing their radiographs with measurements of micromovement between the component and the tibial plateau. Micromovement in the one cemented Miller Galante and seven cementless Ortholoc components, which had been implanted for a range of 3 days to 57 months, was measured with linearly variable differential transducers under anteroposterior shear and axial compressive loads. Micromovement between the tibial component and the screws and between the tibial component and the bone after the screws were removed also was measured in the cementless specimens. Minimal micromovement in most of the implants suggests that the tibial components were well fixed. Micromovement between the tibial tray and the screws also was minimal in the cementless components. Removing the screws did not significantly affect micromovement, except in the specimen retrieved 3 days after surgery and in the specimen with a complete radiolucent line under the component. The cementless tibial components fixed with screws, pegs, and stem were as stable as the component secured with cement. Partial radiolucencies were not associated with greater micromotion than that of bone ingrown areas, but the component with complete radiolucency did have greater micromotion than that of all of the other specimens. PMID- 10379330 TI - Orthopaedic surgeons. Inheritors of tradition. AB - Bonesetting, a most ancient healing art, was absorbed into orthopaedic surgery approximately 120 years ago when Hugh Owen Thomas, a medical doctor specializing in pediatric deformities, assumed his father's nonmedical bonesetting practice. In many nonWestern nations, however, traditional bonesetters continue to treat large numbers of patients. Current market forces in the United States threaten to reduce the role of orthopaedic surgeons in the management of fractures not requiring surgery, leading to the possible reemergence of bonesetting as a separate and independent discipline. PMID- 10379331 TI - Leg pain in an 11-year-old boy. PMID- 10379332 TI - Children with Legg-Perthes diseases. PMID- 10379334 TI - How to manage metastatic brain tumors. PMID- 10379335 TI - Brain metastases in musculoskeletal sarcomas. AB - BACKGROUND: In musculoskeletal sarcomas, brain metastases are rare, but severely affect quality of life. METHODS: All patients with musculoskeletal sarcomas who were treated at our institutions from 1975 to 1997 were reviewed for examples of brain metastasis. RESULTS: Of 480 sarcoma patients, 179 had distant metastases, including 20 patients with brain metastases (4.2%). Alveolar soft part sarcoma (3/4), extraskeletal Ewing's sarcoma (2/8), rhabdomyosarcoma (2/13) and bone Ewing's sarcoma (2/18) tended to metastasize to the brain. All 20 patients had distant or local relapses and 16 of the 20 patients had pulmonary metastases. Three patients underwent surgical treatment and two of them survived over 1 year. Mean survival after diagnosis of brain metastasis was 5.1 months. CONCLUSIONS: Patients with alveolar soft part sarcoma, Ewing's sarcoma, rhabdomyosarcoma and pulmonary metastases have a high risk of brain metastasis. PMID- 10379336 TI - How the lymph node metastases toward cervico-upper mediastinal region affect the outcome of patients with carcinoma of the thoracic esophagus. AB - BACKGROUND: The aim of this study was to establish whether the site of lymph node metastasis influences the survival of patients with carcinoma of the thoracic esophagus. METHODS: A series of 159 patients with lymph node metastasis who underwent right transthoracic R0 esophagectomy was analyzed retrospectively. Sites of the nodal metastasis were divided into two regions; the neck and/or upper mediastinum above (upward metastasis) and the abdomen and/or lower mediastinum below (downward metastasis) the tracheal carina. RESULTS: Univariate analysis of prognostic factors revealed the tumor location, distant lymphatic metastasis, number of metastatic nodes and upward metastasis influenced survival, but downward metastasis did not. Multivariate analysis showed that the number of metastatic nodes and upward metastasis were also significant prognostic factors. Thirty-one (33.3%) of the 93 patients with, but only 6 (9.1%) without, upward metastasis had recurrences in the neck and/or upper mediastinum (P = 0.0002). Eighteen (60.0%) of the 30 patients with extranodal invasion in the neck and/or upper mediastinum had recurrence in these regions. CONCLUSIONS: Nodal metastasis in the neck and/or upper mediastinum was a significant risk factor for prognosis, the same as the number of metastatic nodes. PMID- 10379337 TI - Brain metastases from adenoendocrine carcinoma of the common bile duct: a case report. AB - A 68-year-old man with metastatic brain tumors from adenoendocrine carcinoma of the common bile duct is reported. A common bile duct tumor and a metastatic liver tumor had been resected 6 years and 3 years prior to admission, respectively. Microscopically they showed two components; moderately differentiated tubular adenocarcinoma and neuroendocrine carcinoma. He presented with headache and vomiting and MRI revealed two metastatic brain tumors. They were successfully resected and radiotherapy was carried out. Histological diagnosis of the metastatic brain tumors was neuroendocrine carcinoma, but carbohydrate antigen (CA)-19-9 and carcinoembryonic antigen (CEA)-immunoreactive cells were observed without glandular pattern. Immunohistochemically serotonin and pancreatic polypeptide were detected, but somatostatin was not. As the endocrine cells demonstrated in the normal extrahepatic bile ducts are only somatostatin containing D cells, these cells are considered to originate as part of a metaplastic process. To our knowledge, this represents the second case of adenoendocrine carcinoma of the common bile duct. PMID- 10379338 TI - A large maxillofacial prosthesis for total mandibular defect: a case report. AB - We successfully fabricated a large maxillofacial prosthesis for replacement of a total mandibular defect resulting from surgical failure to reconstruct the mandible. Although a number of reports have described procedures for fabricating midfacial prostheses, there is little information on prostheses to compensate for total loss of the mandible. A 54-year-old woman was referred to the Dentistry and Oral Surgery Division of the National Cancer Center Hospital with total loss of the mandible and the surrounding facial soft tissue. The facial prosthesis we used to treat this patient is unique in that it is adequately retained without the use of extraoral implants and conventional adhesives. This prosthesis is retained by the bilateral auricles and the remaining upper front teeth. We present details of the design of this large silicone maxillofacial prosthesis, with which we successfully rehabilitated the patient. PMID- 10379340 TI - Report of the 1st US-Japan workshop on clinical trials for further development of cancer therapeutics. PMID- 10379339 TI - Retroperitoneal germ cell tumor treated by PVeBV chemotherapy: a case report. AB - The extragonadal germ cell tumor are uncommon neoplasms which account for only 1 5% of germ cell tumors, and its prognosis is poor. We report here the use of combination chemotherapy with cisplatin, etoposide, bleomycin, and vinblastine (PVeBV) for the treatment of retroperitoneal germ cell tumor. A 28-year-old male with complaints of abdominal pain and lumbago, without any abnormality in both testes by physical and ultrasonographic examination, showed retroperitoneal tumor by abdominal computed tomography. The serum alpha-fetoprotein and lactate dehydrogenase were elevated. The retroperitoneal tumor was treated surgically. The pathological diagnosis was mixed germ cell tumor. The lung and supraclavicular lymph node metastases disappeared completely after 3 courses of PVeBV chemotherapy with cisplatin (40 mg/m2 per day) and etoposide (100 mg/m2 per day) for 5 consecutive days, with vinblastine (0.2 mg/kg) on day 1, and bleomycin (30 mg/body) given on days 1, 8, and 15. Granulocyte colony-stimulating factor and serotonin receptor antagonist application were available on acute phase toxic effects. The patient is now alive and well, without recurrence, more than 26 months after the operation. PMID- 10379341 TI - Three-dimensional images by helical CT scan with intra-arterial injection of diluted contrast medium. PMID- 10379342 TI - Practical information on the conduct of randomized trials. An example from The Netherlands. PMID- 10379343 TI - Knowledge grows as it is shared. PMID- 10379345 TI - Japanese recording rule and classification for cancer of various organs. PMID- 10379344 TI - The first transplant from a brain-dead patient in Japan. PMID- 10379346 TI - [Eating habits and cardiovascular risk factors. Epidemiologic study of the Tunisian Sahel]. AB - OBJECTIVES: We assessed carbohydrate, fat and protein calorie intake in the urban population of Sousse, Tunisia in 1996 to evaluate the relationship with cardiovascular risk factors. PATIENTS AND METHODS: A descriptive epidemiology study of a representative sample (n = 957) of the adult population living in the urban areas of Sousse was based on a standardized questionnaire and physical examination (blood pressure, weight, height). Dietitians recorded food intake (previous 24 hours) and eating habits in all participants. In this population, estimated prevalences were: hypertension 18.8%, diabetes 10.2%, obesity 27.7%, android obesity 36%, smoking 21.4%. RESULTS: Total calorie intake was 2483 kcal (67% carbohydrates, 18% protein, 15% fat). There were no significant differences by sex or by presence or absence of cardiovascular risk factors. CONCLUSION: This descriptive study confirmed the epidemiologic transition of the urban population of Sousse, Tunisia. Increased cardiovascular risk results from high, predominantly carbohydrate, calorie intake. It would be advisable to encourage nutritional education on the individual level and the Mediterranean diet on the population level. PMID- 10379347 TI - [Acute adrenal insufficiency due to bilateral adrenal hematoma following severe thrombopenia induced by low-molecular-weight heparin]. AB - BACKGROUND: Bilateral adrenal hematoma is an uncommon cause of acute adrenal insufficiency. An association with thrombopenia induced by low-molecular-weight heparin even more so. Diagnosis is difficult as the clinical manifestations mimic septic shock. CASE REPORT: A 63-year-old woman developed acute adrenal insufficiency due to bilateral adrenal hematoma following severe thrombopenia induced by low-molecular-weight heparin prescribed after an orthopedic operation. Outcome was favorable. CONCLUSION: Acute adrenal insufficiency must be entertained as a possible diagnosis in patients with heparin-induced thrombopenia. PMID- 10379348 TI - [Pasteurella multocida pneumonia in a lupus patient: microbial identification with alveolar lavage fluid on blood culture medium]. AB - BACKGROUND: Pasteurella multocida pneumonia mainly occurs in immunodepressed patients. Microbiological proof is difficult to obtain. CASE REPORT: A 36-year old woman with systemic lupus erythematosus treated with cyclophosphamide and corticosteroids developed pneumonia. She was given amoxicillin-clavulanate. Bronchioalveolar lavage fluid cultures on gelose were negative but Pasteurella multocida grew on blood culture medium. DISCUSSION: Although the direct examination of bronchioalveolar lavage fluid demonstrated Gram negative coccobacilli, gelose cultures were negative, probably because of prior antibiotic therapy. The causal pathogen was only identified when BAL fluid was seeded on blood culture medium, allowing susceptibility tests and subsequent early adaptation of antibiotic therapy. This technique can be helpful in identifying the casual pathogen in microbial pneumonia. PMID- 10379349 TI - [Acute glaucoma in the course of treatment with aerosols of ipratropium bromide and salbutamol]. PMID- 10379350 TI - [Hypoglycemia due to cibenzoline: role of hyperinsulinism]. PMID- 10379351 TI - [Subacute paraneoplastic cerebellar degeneration preceding cerebellar metastasis of ovarian origin. Two rare complications of ovarian cancer]. PMID- 10379352 TI - [Are there criteria which make it possible to predict survival after deep venous thrombosis and pulmonary embolism?]. PMID- 10379353 TI - [Is there a clinical future for echographic measurement of intima media thickness?]. PMID- 10379354 TI - [Follow-up of a patient with type 2 diabetes with the exclusion of follow-up of complications. National Agency of Accreditation and Evaluation of Health]. PMID- 10379355 TI - [Follow-up of the type 2 diabetic patient. An interview with B. Charbonne. UK Prospective Diabetes Study Group]. PMID- 10379356 TI - [Evaluation of the quality of life in children and adolescents]. AB - HEALTH-RELATED QUALITY OF LIFE: Personal perception of health is proposed as an innovating assessment alternative for making medical and therapeutic choices on an individual and population basis. Standardized questionnaires have been developed to assess health-related quality of life (HRQL). ASSESSMENT OF HRQL IN ADOLESCENTS AND CHILDREN: Several approaches have been used. Some questionnaires assess health status in specific diseases or populations. More general instruments developed for the general population are useful for comparing between programs involving different diseases. PRACTICAL APPLICATIONS: We present here the criteria of quality required for HRQL instruments and examine the difficulties encountered when applying these instruments to children and adolescents. We reviewed the main general instruments in the literature as well as those specifically designed for asthmatic children. None of the self administered questionnaires available in France are fully satisfactory for children or adolescents in terms of robustness, sensitivity and discriminative power. The French versions of the Childhood Asthma Questionnaire developed in Great Britain and the Paediatric Asthma Quality of Life Questionnaire developed in Canada were tested in a transcultural validation scheme. PMID- 10379357 TI - [Satisfaction with anesthesia: a review of the existing instruments]. AB - EVALUATING PATIENT SATISFACTION: In France, patient satisfaction is a criteria for health care facility accreditation. In this context, the anesthesia community has studied tools available for assessing satisfaction with anesthesia. LIMITATIONS OF AVAILABLE INSTRUMENTS: The construction of a satisfaction assessment instrument can be divided into three phases: design, sorting items, validation. Instruments available in the currently literature (1987-1997) focus mainly on pre, per- or postoperative management but little on overall patient satisfaction. As the concept of satisfaction concerns a variety of elements, many of the available instruments use a multidimensional approach. The areas explored however vary greatly depending on the author or the study. In addition, patient experience is rarely taken into consideration when designing instruments. FOUR INSTRUMENTS: Among the available instruments we retained 4 questionnaires: Patient Satisfaction with General Anaesthesia, Peri-operative Anesthesia Experience Scale, Iowa Satisfaction with Anesthesia Scale, and Amerstadam Preoperative Anxiety and Information Scale. These last 2 scales only assess patient information and anxiety. The Peri-operative Anesthesia Experience Scale alone is available in French (Echelle de Vecu perioperatoire de l'Anesthesie). Patient satisfaction is also approached with specialized instruments designed to assess specific items. The most widely used scales assess anxiety and pain. Based on these findings, it is dear that the fundamental concept of patient satisfaction must be revisited. We propose a few points for thought. PMID- 10379358 TI - Genetics and molecular pathogenesis of mitochondrial respiratory chain diseases. AB - Dysfunction of the mitochondrial respiratory chain has been recognised as a cause of human disease for over 30 years. Advances in the past 10 years have led to a better understanding of the genetics and molecular pathogenesis of many of these disorders. Over 100 primary defects in mitochondrial DNA (mtDNA) are now implicated in the pathogenesis of a group of disorders which are collectively known as the mitochondrial encephalomyopathies, and which most frequently involve skeletal muscle and/or the central nervous system. Although impaired oxidative phosphorylation is likely to be the final common pathway leading to the cellular dysfunction associated with such mtDNA mutations, the complex relationship between genotype and phenotype remains largely unexplained. Most of the genes which encode the respiratory chain reside in the nucleus, yet only five nuclear genes have been implicated in human respiratory chain diseases. There is evidence that respiratory chain dysfunction is present in common neurological diseases such as Parkinson's disease and Huntington's disease. The precise cause of this respiratory chain dysfunction and its relationship to the disease process are unclear. This review focuses upon respiratory chain disorders associated with primary defects in mtDNA. PMID- 10379359 TI - SNAREs and SNARE regulators in membrane fusion and exocytosis. AB - Eukaryotes have a remarkably well-conserved apparatus for the trafficking of proteins between intracellular compartments and delivery to their target organelles. This apparatus comprises the secretory (or 'protein export') pathway, which is responsible for the proper processing and delivery of proteins and lipids, and is essential for the derivation and maintenance of those organelles. Protein transport between intracellular compartments is mediated by carrier vesicles that bud from one organelle and fuse selectively with another. Therefore, organelle-specific trafficking of vesicles requires specialized proteins that regulate vesicle transport, docking and fusion. These proteins are generically termed SNAREs and comprise evolutionarily conserved families of membrane-associated proteins (i.e. the synaptobrevin/VAMP, syntaxin and SNAP-25 families) which mediate membrane fusion. SNAREs act at all levels of the secretory pathway, but individual family members tend to be compartment-specific and, thus, are thought to contribute to the specificity of docking and fusion events. In this review, we describe the different SNARE families which function in exocytosis, as well as discuss the role of possible negative regulators (e.g. 'SNARE-masters') in mediating events leading to membrane fusion. A model to illustrate the dynamic cycling of SNAREs between fusion-incompetent and fusion competent states, called the SNARE cycle, is presented. PMID- 10379360 TI - The molecular basis and clinical aspects of Peutz-Jeghers syndrome. AB - Peutz-Jeghers syndrome (PJS) is a classic, but not widely known hereditary trait [1, 2]. Its clinical hallmarks are intestinal hamartomatous polyposis and melanin pigmentation of the skin and mucous membranes. In addition, PJS predisposes to cancer [3, 4]. The most common malignancies are small intestinal, colorectal, stomach and pancreatic adenocarcinomas. Other cancer types that probably occur in excess in PJS families include breast and uterine cervical cancer, as well as testicular and ovarian sex cord tumors. The relative risk of cancer may be as high as 18 times that of the general population, and the cancer patients' prognosis is reduced. Recently, the predisposing locus was mapped to 19p13.3 using a novel method [5]. Subsequently, the causative gene was shown to be LKB1 (a.k.a. STK11), a serine/threonine kinase of unknown function [6]. Although preliminary reports seem to suggest a minor role for LKB1 in sporadic tumorigenesis [7-12], further investigations are needed. PMID- 10379361 TI - Immune responses to DNA vaccines. AB - DNA vaccines, based on plasmid vectors expressing an antigen under the control of a strong promoter, have been shown to induce protective immune responses to a number of pathogens, including viruses, bacteria and parasites. They have also displayed efficacy in treatment or prevention of cancer, allergic diseases and autoimmunity. Immunologically, DNA vaccines induce a full spectrum of immune responses that include cytolytic T cells, T helper cells and antibodies. The immune response to DNA vaccines can be enhanced by genetic engineering of the antigen to facilitate its presentation to B and T cells. Furthermore, the immune response can be modulated by genetic adjuvants in the form of vectors expressing biologically active determinants or by more traditional adjuvants that facilitate uptake of DNA into cells. The ease of genetic manipulation of DNA vaccines invites their use not only as vaccines but also as research tools for immunologists and microbiologists. PMID- 10379362 TI - Uteroglobin: a novel cytokine? AB - Blastokinin or uteroglobin (UG) is a steroid-inducible, evolutionarily conserved, multifunctional protein secreted by the mucosal epithelial of virtually all mammals. It is present in the blood and in other body fluids including urine. An antigen immunoreactive to UG antibody is also detectable in the mucosal epithelia of all vertebrates. UG-binding proteins (putative receptor), expressed on several normal and cancer cell types, have been characterized. The human UG gene is mapped to chromosome 11q12.2 13.1, a region that is frequently rearranged or deleted in many cancers. The generation of UG knockout mice revealed that disruption of this gene causes: (i) severe renal disease due to an abnormal deposition of fibronectin and collagen in the glomeruli; (ii) predisposition to a high incidence of malignancies; and (iii) a lack of polychlorinated biphenyl binding and increased oxygen toxicity in the lungs. The mechanism(s) of UG action is likely to be even more complex as it also functions via a putative receptor mediated pathway that has not yet been clearly defined. Molecular characterization of the UG receptor and signal transduction via this receptor pathway may show that this protein belongs to a novel cytokine/chemokine family. PMID- 10379363 TI - Minoxidil-induced cardiac hypertrophy in guinea pigs. AB - To investigate whether during cardiac hypertrophy changes occur in contractile protein composition and in mechanical and energetic properties of the myocardium, contractile protein composition, isometric force and adenosine triphosphate (ATP) consumption were studied in control and hypertrophied guinea-pig hearts. Cardiac hypertrophy was induced by adding minoxidil (120 or 200 mg/l) to the drinking water. Protein analysis was performed by one-dimensional gel electrophoresis. The myosin heavy-chain (MHC) composition was determined in an enzyme-linked immunosorbent assay (ELISA). ATP consumption and force development were simultaneously measured during isometric contraction in chemically skinned trabeculae. Histochemical analysis of cross-sectional area of cardiomyocytes and interstitial space was performed on the left ventricular tissue of 200 mg/l minoxidil-treated and control guinea pigs. Minoxidil treatment (120 and 200 mg/l) significantly increased left ventricular dry weight normalized for body weight by 19 +/- 4 and 24 +/- 4%, respectively. No significant differences were found in the cellular cross-sectional area, while interstitial space was slightly decreased in minoxidil-treated hearts. In left ventricular trabeculae of 200 mg/l minoxidil-treated guinea pigs, ATPase activity was slightly less than in those of control guinea pigs, whereas force did not differ significantly. Calcium sensitivity of force and ATPase activity were not affected by minoxidil treatment. Gel electrophoresis revealed no difference in contractile protein composition, but a tendency towards a lower amount of alpha-MHC in the minoxidil treated hearts was found in ELISA. PMID- 10379364 TI - The role of hsp70 in protection and repair of luciferase activity in vivo; experimental data and mathematical modelling. AB - The stably transfected rat cell line HR24 expressing high levels of the inducible human hsp70 and its parental cell line Rat-1 were used for in vivo studies to analyse the role of hsp70 during thermal protein denaturation and the subsequent renaturation. In order to monitor denaturation and renaturation of a cellular protein in vivo, both cell lines were transiently transfected with firefly luciferase (Luc). The continuous monitoring of Luc activity during and after heat stress allowed a detailed analysis of the inactivation and reactivation kinetics in cells grown in monolayers. The aim of these studies was to distinguish a protective effect of increased hsp70 levels during heat shock-induced protein inactivation from a stimulation of reactivation. In this paper we show that in cells that are stably transfected with hsp70, thermal Luc inactivation decreased, and subsequent reactivation yielded higher activity levels, compared with the parental cells. The difference in early inactivation kinetics observed in the two cell lines suggests an immediate effect of the presence of an extra amount of hsp70 on enzyme inactivation. Using different mathematical models, the heat induced inactivation and reactivation kinetics was compared with simulations of denaturation and renaturation. It is concluded that the model in which it is assumed that hsp70 is able to interact with partially denatured proteins, which did not yet lose their enzymatic activity, most optimally explains the experimental observations. PMID- 10379366 TI - Neural network correction of ultrasonic C-scan images. AB - A neural network-based approach to the correction of C-scan images is presented. This allows the effects of a finite ultrasonic beam diameter in an immersion experiment to be considered, by training the network on a series of defects of known characteristics. The result is an image which is a better representation of the actual defect. PMID- 10379365 TI - Two new aminopeptidases from Ochrobactrum anthropi active on D-alanyl-p nitroanilide. AB - Two new enzymes which hydrolyse D-alanyl-p-nitroanilide have been detected in Ochrobactrum anthropi LMG7991 extracts. The first enzyme, DmpB, was purified to homogeneity and found to be homologous to the Dap protein produced by O. anthropi SCRC C1-38 (ATCC49237). The second enzyme, DmpA, exhibits a similar substrate profile when tested on p-nitroanilide derivatives of glycine and L/D-alanine, but the amounts produced by the Ochrobactrum strain were not sufficient to allow complete purification. Interestingly, the DmpA preparation also exhibited an L aminopeptidase activity on the tripeptide L-Ala-Gly-Gly but it was not possible to be certain that the same protein was responsible for both p-nitroanilide and peptide hydrolysing activities. The gene encoding the DmpA protein was cloned and sequenced. The deduced protein sequence exhibits varying degrees of similarity with those corresponding to several open reading frames found in the genomes of other prokaryotic organisms, including Mycobacteria. None of these gene products has been isolated or characterised, but a tentative relationship can be proposed with the NylC amidase from Flavobacterium sp. K172. PMID- 10379367 TI - Fusional anomalies of the testis and epididymis. AB - Earlier, the cause of infertility in undescended testis (UT) had been widely accepted as a consequence of the higher temperature of the inguinal/abdominal region. Observations made in the past two decades, however, gave new evidences. The most important of these is that UT is often associated with the fusional anomalies (FA) of the testis and epididymis. FA is the consequence of pathological intrauterine hormonal processes and many authors believe FA to be the primary cause for infertility in UT. Since 80% of UT cases are of endocrine origin, it would be suspected that the very same factors are responsible for both UT and FA. FA and other anomalies of the epididymis often occur in testicular torsion (TT) as well. It is remarkable that infertility could follow the unilateral forms of UT and TT despite the presence of a "healthy" contralateral gonad. In both entities contralateral FA (probably associated with testicular dysgenesis) is suspected. These observations could influence the primary surgical treatment of patients with UT and TT, as well as the mode of further management of these cases. PMID- 10379368 TI - Studies on tumourous and other urogenital patients with respect to antigens of oncogenic adenovirus. AB - The possible connection of viruses with tumours was investigated by serologic examinations. Concerning the presence of antibodies against adenoviruses, especially those against the early non-virion antigen of oncogenic adenovirus type 12, approximately 4000 tests were made with sera of 446 urogenital patients with and without tumours and 70 ones with internal diseases. It was found by complement fixation tests that antibodies against non-virion antigens of adenoviruses were present in 53% of urogenital patients suffering from malignant tumours and prostatic hypertrophy, in 18% of non-tumourous urological patients and in 4% of patients with internal diseases, respectively. The results suggest that adenoviruses may play a role in tumourous diseases of the urogenital organs. PMID- 10379369 TI - Comparison of propofol-fentanyl or midazolam-fentanyl intravenous anaesthesia for carotid endarterectomy. AB - Carotid endarterectomy has become a standard surgical operation in the therapy of cerebrovascular insufficiency. The cardiovascular status of the patients needs special attention, since the long-term prognosis is predominantly influenced by concomitant coronary artery disease. General anesthesia techniques are raising the challenge of maintaining cardiovascular stability and establishing adequate cerebral monitoring. Randomly selected 30 patients gave informed consent to this approved study. Fifteen patients were anaesthetised with propofol-fentanyl or midazolam-fentanyl combined with N2O-O2. Haemodynamic parameters (mean arterial pressure, heart rate) showed not significant changes during anaesthesia. Recovery profil proved to be significantly better after propofol-fentanyl compared to midazolam-fentanyl anaesthesia. PMID- 10379370 TI - Non-surgical treatment of erectile dysfunction. AB - Authors survey the conservative treatment possibilities of erectile dysfunction. With the increase in interest, they estimate the new oral- and intracavernosal drugs, giving a review of their experiences with the treatment. A perspective prognosis is given of the important role of NO (nitric oxide) donors and PHD (phosphodiesterase) inhibitor drugs in the field of conservative therapy, while intracavernosal and intraurethral prostaglandin therapies are still important ingredients in the therapeutic arsenal, as is hormone substitution in indicated cases. PMID- 10379371 TI - Testicular sperm aspiration (first Hungarian results). AB - Authors summarize the first results with the use of spermatozoas retrieved with direct surgical method in intracytoplasmic injection in Hungary (1995-1997). Eighty-nine procedures were performed in 65 patients and 84 cases were successful. Out of 84 cases 23 clinical pregnancies could be achieved (27.3%). Thirteen children were born, including one case of twins and one triplets. PMID- 10379372 TI - Laparoscopic varicocele operation: a chance to prevent the recurrence. AB - There are recurrences and postoperative hydrocele of varicocele after any kind of surgical treatment. Laparoscopic clipping and dissection of internal spermatic vessels was performed without any complication in 73 children to treat varicocele in our department between 1995 and 1998. We have used a new method to detect etiological factors at laparoscopic surgery. The well-known Linton and Trendelenburg test was adapted to detect incidental collateral veins in 73 patients. Using these test, collateral veins were detected in 16 boys. The testicular artery identified in most of the cases as a pulsatile vessel. The operating time was 10-25 minutes. Laparoscopic varicocelectomy is a safe, effective treatment causing minimal discomfort and allowing patients an early to return to activity. These results suggest this technique a viable alternative to open ligation in paediatric urological practice. PMID- 10379373 TI - Our experience with the therapy of bleeding peptic ulcers. AB - Authors present their clinical experience gained in the therapy of bleeding peptic ulcers. The modified Baylor Bleeding Score has proved to be a reliable tool for assessing the risk of recurrent hemorrhage. Early introduction of H2 receptor blocker therapy is justified both in conservative as well as in surgical management of bleeding peptic ulcers. PMID- 10379374 TI - Referral patterns and motivation for anti-reflux surgery of patients suffering from gastroesophageal reflux disease. AB - Due to a better understanding of the pathophysiology of gastroesophageal reflux disease (GERD), as well as the improvements in surgical technique, the number of anti-reflux procedures has increased world-wide during the last decade. This trend has been facilitated by the advent of minimally invasive surgery. Although a great number of publications deal with the indications or selection of patients for surgery, only a few discuss the motivation of patients for choosing surgery rather than long-term medical treatment. In order to evaluate the different elements of motivation of patients suffering from primary gastroesophageal reflux disease, the data of 115 patients who had undergone anti-reflux surgery between January 1990 and June 1997 at the Department of Surgery, Technical University, Munich, were evaluated. As laparoscopic anti-reflux surgery has only been regularly performed since 1994 in our Department, the study period was divided and the two periods (1990-1993 and 1994-1997) were analyzed separately. The data were evaluated according to the referral pattern and the motivation of patients with GERD who chose surgery. In the period from 1990 to 1993, 38.5% of the patients were referred to surgery by general practitioners, 38.5% by internists, 10% by practicing surgeons and 8% by gastroenterologists. In 5% of the cases the patients themselves initiated surgery. The corresponding results for the period from 1994 to 1997 were 29%, 38%, 12%, 8% and 13%, respectively. The most common reason for a patient to choose surgery was moderate or only short-term response to appropriate conservative treatment, which accounted for 98% and 92% of the patients, respectively, during the study periods. This was followed by avoidance of life-long medical therapy and its potential risks in 77% and 85% of the patients. Fear of cancer was reported in 10% and 25%, respectively. In the second period, the option of a minimally invasive procedure was reported as an important factor in 45% of the patients. Although the number of anti-reflux procedures performed per year is increasing and there is also an increasing tendency regarding the application of minimally invasive procedures, the factors leading to referral failed to show significant differences in the two periods. The motivation of patients, however, clearly changed in favour of surgical therapy, mainly because of the availability of a minimally invasive approach. PMID- 10379375 TI - Development of a surface modified silicone-keratoprosthesis with scleral fixation. AB - BACKGROUND: Many attempts have been made to create artificial corneas. The keratoprostheses currently available do not allow measurements of the intraocular pressure (IOP) and restrict the visual field. The main problem is extrusion due to an insufficient connection between implant and surrounding tissue. It is our aim to create a flexible keratoprosthesis with a wide field optic allowing measurements of the IOP. Surface modification will improve cell adhesion and therefore stability between implant and tissue. METHODS: The keratoprosthesis is made of silicone rubber. The optical zone is 11 mm in diameter with a thickness of 0.3 mm. The surface modified haptic consists of a scleral rim and 8 branches for scleral fixation. Optical and mechanical qualities were tested by tensile tests, spectrophotometry and topography. RESULTS: A method to produce one-piece silicone keratoprostheses was established. Submicron lathing of the mould led to an excellent optical quality. Spectrophotometry showed high degree of visible and ultraviolet light transmission of the silicone. Mechanical tests revealed high tensile strength and elongation at break which were not impaired by surface modification. CONCLUSION: The production of a flexible silicone keratoprosthesis with high optical and mechanical properties was accomplished, with possible use as both permanent and temporary keratoprosthesis. PMID- 10379376 TI - Asymptotic parathyroid cyst--diagnostic difficulties (a case report). AB - The case of a 70-year-old female with asymptotic parathyroid cyst is presented. It was not possible to make a proper diagnosis preoperatively. The correct diagnosis was based on the result of the postoperative microscopic examination of the resected tissues. The diagnostic difficulties are discussed. PMID- 10379377 TI - Experiences of 25 orthotopic ileal neobladders. AB - Authors report on their results obtained from orthotopic ileal neobladders following 25 cases of radical cystectomy. Analysis is given of the possible complications, their prevention, as well as of surgical techniques. It is determined that orthotopic ileal neobladder is one of the best bladder substitution methods, giving the patient a chance for a high quality of life. PMID- 10379378 TI - Experiences on 25 cases of radical cystectomy. AB - Authors review their early experiences and the oncopathological relations in respect to 25 cases of radical cystectomy involving orthotopic bladder substitution. The difficulties of diagnostics and indication are discussed. Attention is drawn to the fact that pathological "staging", "grading" are not entirely exact and reliable, though surgical indication is positioned on these. It is the opinion of authors that in case of TIG3 radical surgery is indicated. Due to the shortness of the follow-up periods, no studies on survival were performed. It remains an open question whether radical cystectomy is indicated as opposed to the possible choice of organ-preserving surgery. PMID- 10379379 TI - Factors affecting prognosis of renal cell carcinomas. AB - Author analyzed the survival of 126 patients operated on because of kidney carcinomas. The data on anamnesis, laboratory results and TNM histological classification were compared with the results on "relative survival" and correlation analysis was performed. Based on these, the 12 most important criteria (with the highest correlation coefficient) were emphasized and regression equation was formed based on the TSP--Time Series Processing--Version 4 (Hall). Using this method, a numerical prognostic index can be established for the judgement of prognosis regarding certain patients. PMID- 10379380 TI - Schistosomiasis of the urinary bladder. AB - Authors review a case of urinary schistosomiasis, where the process caused left sided urinary obstruction. Because of the suspicion of a tumor transurethral resection was performed, whereafter the ureter passage became unhindered. Diagnosis became clear upon the histological examination of the resected tissue. Based on the cited literature, reference is made to the mortality rate of the disease in the expanded endemic areas, as well as to the high number of patients at risk. A brief summary is given of the pathology, symptoms, diagnostics of the disease, with mention of differentiation and therapeutic possibilities as well. With regard to importation of the disease, attention is called to the importance of careful anamnesis. PMID- 10379381 TI - Retroperitoneal malignant fibrous histiocytoma. AB - Authors review the case history and follow-up a rare malignant fibrous histiocytoma patient, based on the relevant literary data. The tumor filled the retroperitoneum on the right side, in front of the right kidney. Intravenous urography and computer tomography revealed a 10 x 15 cm sized mass, suspect of being a kidney tumor. Upon surgery, the tumor was found to be a retroperitoneal malignant fibrous histiocytoma. In connection to the case, a brief review is given of the storiform type of malignant fibrous histiocytoma, regarding its etiological, clinical and pathological aspects, the difficulties in diagnosis, as well as the therapeutic possibilities. Authors regard their case worthy of publication because of the retroperitoneal location and significant size of the tumor, and because of the unproven diagnosis prior to surgery. Even after 4 years the patient is symptom- and complaint-free, and CT has revealed no metastases. PMID- 10379382 TI - Recent results of fungal taxonomy. PMID- 10379383 TI - Extracellular proteins in fungi: a cytological and molecular perspective. AB - Protein secretion is a vital process in fungi. For many, the secretion of hydrolytic enzymes provides a crucial step in their nutrition in nature. However, in recent years the list of different types of secreted proteins that have been discovered has extended significantly. These have been shown to have a diversity of functions including toxic molecule transport and control of desiccation. The majority of secreted proteins are glycosylated and our understanding of this aspect of fungal biochemistry has also extended in recent years. This review addresses the process of protein secretion from the cytological, biochemical and genetical standpoints. Advances in technology in many areas of scientific approach have enabled a better and understanding of this important process in fungi. PMID- 10379384 TI - Origins, phylogenies and relationships in the fungal Kingdom. PMID- 10379385 TI - Molecular taxonomy of yeasts. AB - In the last two decades the application of molecular techniques has had a major impact on the classification of yeasts. The nuclear DNA relatedness has become the basis of species delineation. Molecular fingerprinting methods such as analysis of restriction fragment length polymorphisms, random amplified polymorphic DNA, PCR-amplified sequences and fragments, pulsed field gel electrophoresis of chromosome DNA and others allow intraspecies differentiation and typing. The most far reaching method has been the sequencing of various parts of ribosomal DNA that has made for the first time possible to assess the phylogenetic relationships among yeasts at different taxonomic levels. Based on the molecular data obtained so far several changes have been introduced in the classification of yeasts, however, substantial restructuring of current taxonomic schemes with the consequence of numerous nomenclatural changes must await further studies. PMID- 10379386 TI - Questions of classification of basidial fungi. AB - Surveying mycological works dealing with great number of species about basidial fungi published in the last hundred years (Kalchbrenner [1], Istvanffi [2], Hollos [3], Moesz [4, 5], Bohus et al. [6], Banhegyi et al. [7], Ubrizsy [8], Banhegyi et al. [9], Babos [10, 11], Rimoczi-Vetter [12], Rimoczi [13]) the great variety and changes of applied systems is conspicuous. In all works the doubt is expressed whether the currently applied systematic lists and nomenclatural solutions are the best, and whether they have chosen the most appropriate systematic theories. Hungarian authors have never created an own system though it is conspicuous that their knowledge about foreign mycologist taxonomist's works of the given period is thorough. Critical usage of their elements and critical review in the own works is rare (Ubrizsy-Voros [14], Ubrizsy [15]). PMID- 10379387 TI - Fungal symbioses. PMID- 10379388 TI - The role of fungi in the carbon- and nitrogen cycles. PMID- 10379390 TI - From edible to useful mushrooms--an attempt for the new economical assessment of large fungi. AB - According to the stand of the modern applied mycological research the most commonly used term "edible mushroom" does not express all significant aspects large fungi can be used for. Additionally to bioconversion for food and animal feed production there are at least three other fields where large fungi may also get economical relevance: for establishing of ectomycorrhiza, for medical application and for soil decontamination including environmental engineering. This new situation justifies the introduction of a new, all-embracing designation for large fungi. The term "useful mushroom" will be suggested. The various options of the use of mushrooms will be introduced and briefly discussed in this paper. PMID- 10379389 TI - The research of lichenized fungi in Hungary. AB - A brief historical account on the Hungarian lichenology is given in three stages. The early stage involves the oldest collections and the lichenological activity of F. Hazslinszky. In the mid-stage a separate, independent lichen collection was established in the Natural History Museum, and a rapid development of floristical and taxonomic research took place. Recent lichenological research in Hungary follows several fields: taxonomy, biogeography, bioindication and ecophysiology. PMID- 10379391 TI - Production of bioactive compounds by different fungal species. PMID- 10379393 TI - Current trends in medical mycology. PMID- 10379392 TI - Hygienic mycology in Hungary. AB - In Hungary the medical mycological research concerning the systemic (so-called deep) mycoses started in the National Institute of Hygiene. The Mycology Department has been founded in 1955 in this Institute with nation-wide authority as a routine diagnostic laboratory for clinical specimens from systemic mycotic infections. Three years later its activity was broadened--on ground of the experiences--to cover the total field of hygiene, thus we had to establish the "mycologia hygienica" (hygienic mycology), however, this has not been accepted as an independent discipline. Here a short, selected overview is given of our experiences. In the following the trends of the activity is demonstrated along the subdivisions of hygiene (epidemiology, environmental-hygiene, professional and occupational-, nutritional and food hygiene). In respect of hygiene, however, the fungi play a Janus-faced (more exactly Dr. Jekyll-Mr. Hyde) role, as they can behave not only harmful but also beneficial. PMID- 10379394 TI - Changing epidemiology of systemic fungal infections and the possibilities of laboratory diagnostics. PMID- 10379395 TI - Recent progress in mycotoxin research. PMID- 10379396 TI - Fungi in forestry--significance and research in Hungary. AB - The importance of the fungi in forest ecosystems and in forestry as well as a short review of the research of fungi with forestry significance in Hungary are discussed in the paper. PMID- 10379397 TI - Current research on phytopathogenic fungi: an overview. PMID- 10379398 TI - Biological control of agricultural pests by filamentous fungi. PMID- 10379399 TI - Signal transduction in fungi--the role of protein phosphorylation. PMID- 10379400 TI - Chromosomes, karyotype analysis, chromosome rearrangements in fungi. AB - In this review the organization of fungal chromosomes and the methods used for karyotype analysis are briefly summarized. The role of chromosome rearrangement, supernumerary chromosomes and repeated DNA sequences in the genetic change of fungi is evaluated. PMID- 10379401 TI - Extrachromosomal genetic elements in fungi. PMID- 10379402 TI - Impact of yeast genetics and molecular biology on traditional and new biotechnology. AB - Developments in yeast genetics, biochemistry, physiology and process engineering provided bases of rapid development in modern biotechnology. Elaboration of the recombinant DNA technique is far the most important milestone in this field. Other molecular genetic techniques, as molecular genotyping of yeast strains proved also very beneficial in yeast fermentation technologies. Saccharomyces cerevisiae is the most exploited eukaryotic microorganism in biotechnology but non-Saccharomyces species are becoming more and more important in the production of perfectly translated heterologous proteins. PMID- 10379403 TI - Genetics, physiology and cytology of yeast-mycelial dimorphism in fission yeasts. AB - The order Schizosaccharomycetales contains a dimorphic and two yeast species. Sch. japonicus can form both yeast cells and mycelium, depending on the substrate and the culturing conditions. Sch. pombe is a strictly unicellular organism, but it can be forced to form mycelial cell chains by inactivating members of the sep gene family. The mutations in most of the sep genes confer pleitropic phenotypes indicating functional involvement in MAP-kinase-mediated signalling pathways. Two of them were found to encode transcription factor homologues of other eukaryotes. PMID- 10379405 TI - [In Process Citation] PMID- 10379404 TI - Industrial microbiologists in the area of microfungi. PMID- 10379406 TI - [Allergens]. AB - The clinical signs linked with immediate hypersensitivity, correspond with an abnormal reactivity of the immune system, which appears following contact with external substances, the allergens. Over the last few years there has been considerable progress in research on these substances in the domain of structural characterisation, biochemistry and immunological properties. Besides the definition of the word allergen linked to its immunological characteristics, our usual language maintains a certain ambiguity in its use which may characterise different states, including successive steps of the manufacture of an allergen extract. In effect the word allergen may designate the agent that is responsible for the allergic disease, for example cat, but also the raw material used for the manufacture of the corresponding extract, whether it be hair or squames; it may also apply to the final allergenic extract, the extract of cat hair or squames, as well as a precise molecule such as the major allergen of cat Fel d1 in the same example. After having reviewed several definitions as well as the nomenclature we will study the general characteristics of pneumoallergens and trophallergens, those of recombinant allergens, then the parameters of manufacture of allergen extracts. PMID- 10379407 TI - [Biological assays in allergology: constraints, concepts, criteria, and methods of evaluation]. AB - The in vitro diagnosis of allergies is submitted to several constraints some shared by all the in vitro methods and some which are specific of immuno allergology. The in vitro diagnosis of allergies involves various parameters among which the allergen by itself is far the most complex. On the basis of a studied population having an optimal size, the most powerful statistical parameters are the positive predictive value, the negative predictive value and the efficiency. The bio-clinical correlations lead to the involvement of another parameter which is the prevalence of the clinical reaction which may be corrected by a clear clinical history. Finally, the absence of absolute gold standard in allergology lead to the conclusion that allergy diagnosis is still based on the synthesis of various clinical and biological arguments. PMID- 10379408 TI - [Apropos of the evaluation and validation of in vitro tests for measurement of specific IgE]. AB - The performances of the reagents for IN VITRO TESTS in Allergy, are very variable, in spite of the existence of registration procedures as well as those of national quality control. Most of the discrepancies have their origin in the choice of technologies used by the manufacturers (reaction support, raw materials, calibration, vigour of the process of execution). It follows from this that the characteristics must be understood and the limitations appreciated of the tests that are used, as well as their interpretation. Of such information that is accessible in the records of the National Control of Quality of the Medicines Agency, as well as in the studies that are done with the aim of evaluating analytical and/or clinical performance of available tests (this work is a review of the performances of most of the tests sold in France today). It stands out that the best borders on very mediocre and that it is indispensable, like the planned nomenclature of the medical biological consultations, to take into consideration the technique used to obtain a result for interpretation of the latter. PMID- 10379409 TI - ["Allergic Progression": importance of Phadiatop and RAST fx5 as screening measures for childhood atopy]. AB - Since discovery of IgE in 1967, which was a real revolution in allergy field, new biological methods as Phadiatop for inhalant allergens and RAST fx5 for food allergens, have been used routinely. Both methods are very useful for reliable and cost efficient atopy screening. PMID- 10379410 TI - [Immunobiology of respiratory allergy]. AB - Respiratory allergies are frequent and the etiological diagnosis is not always easy. Most often, the allergy assessment is vital. For the allergist, this assessment begins with skin tests which must be complemented by measurement of the level of specific IgE in case of doubt. For the generalist or physician who does not have special competence in allergy, an alternative to a specialist consultation at once is a multi-allergen test of detection. This assessment is necessary to set out the etiological diagnosis and facilitate the responsibility of the therapy. PMID- 10379411 TI - Verb naming in normal aging. AB - Few studies have examined verb naming in normal aging, although decline in the ability to name nouns has been well documented. In this study, we examined longitudinal performance on the Action Naming Test (ANT), a confrontation naming test for verbs. The purpose of this study was to confirm the verb naming deficit associated with aging, which was previously seen only in cross-sectional studies, and to provide additional normative data on verb naming ability that may prove useful to studies on verb naming in populations with brain dysfunction. Sixty-six healthy men and women aged 30 to 79 were each tested with the ANT 3 times over a 7-year span. ANT performance showed a significant decline over time for all participants except the youngest group. Longitudinal methodology supports the conclusion that this finding of a decline in verb naming ability arises from true age-related changes and not from cohort differences. PMID- 10379412 TI - Adult reading assessment: are we doing the best with what we have? AB - This article describes a survey completed by 728 neuropsychologists for the purpose of gathering information about the assessment of reading in adults as part of neuropsychological examinations. The survey information gathered addressed (a) the general frequency of assessing adult reading, (b) the assessment tools used, (c) the general purposes for the assessment of reading, (d) the need for a review describing available adult reading norm-referenced tests, and (e) the need for the development of criterion-referenced reading tests appropriate for determining functional reading abilities. Survey findings are reported and discussed. A list and description of reading tests appropriate for assessing reading in adults also is provided in the Appendix. PMID- 10379413 TI - Raw, demographically altered, and composite Halstead-Reitan Battery data in the evaluation of adult victims of nonimpact acceleration forces in motor vehicle accidents. AB - Data from the Halstead-Reitan Neuropsychological Test Battery were interpreted for 33 adults who had been subjected to nonimpact acceleration forces in motor vehicle accidents. Comparisons with normative data provided by Reitan and Wolfson (1993) identified impaired performance on the Localization component of the Tactual Performance Test and the Category Test and atypical right-left differences on the Tactual Performance Test and Grip Strength. In contrast, comparisons with a normative data set developed by Heaton, Grant, Matthews, and PAR Staff (1991) through T-score conversions produced results that suggested normal cognitive skills. The Neuropsychological Deficit Scale score was one of the clearest indicators of neuropsychological impairment, falling generally well within the mildly impaired range. PMID- 10379414 TI - Performance of psychotic and substance abuse patients with or without head injury on the Halstead-Reitan Battery. AB - Performances of patients diagnosed with psychosis or substance abuse disorder were evaluated using the Halstead-Reitan Neuropsychological Test Battery (Reitan & Wolfson, 1993) and the General Neuropsychological Deficit Scale (GNDS). Half of the participants in each group had a documented head injury that required medical attention. Differences between patient groups with head injury was not significantly above those without head injury on the Halstead-Reitan subtests or the GNDS. Average performance in all 4 groups was in the mild to moderate impairment range according to established norms and, depending on the group, with 80% to 93% of patients scoring above the accepted cutoff for impairment on the GNDS. This study has implications for referrals attempting to distinguish head injury effects from those of psychosis or substance abuse. The GNDS did not statistically differentiate head injury effects but in these populations did detect neurological impairment, which supports the utility of this index as an indicator of overall neurological impairment. PMID- 10379415 TI - Memory and metamemory functioning among depressed patients. AB - Memory and metamemory functioning were studied among 30 adult patients suffering from major depression. The results indicate that, besides showing signs of cognitive slowing, the patients were especially vulnerable to visual memory impairment, whereas verbal, short-term memory, and recall by recognition were more often unaffected. The patients whose depression was characterized by physiological symptoms, such as loss of appetite and sleep disturbances, showed impairment in traditional short-term memory measures, whereas there was no firm connection between cognitive or behavioral depressive symptoms and memory functioning. The depressive patients' generalized view of their memory capability was strongly underestimated, whereas online metamemory accuracy by which one perceives and makes inferences about one's performance was adequate. PMID- 10379416 TI - Longitudinal neuropsychological evaluation of a case of pineal tumor occurring in an adolescent girl. AB - There has been a relative absence of studies that have longitudinally examined the neuropsychological profiles of patients suffering from pineal tumors. A case is reported of an adolescent girl with a pineoblastoma and spinal metastases who received extensive chemotherapy and cranio-spinal irradiation. Neuropsychological assessments conducted approximately 5 months and 2.5 years posttumor diagnosis revealed a diversity of impairments indicative of mild to moderate neuropsychological dysfunction. By the 2nd evaluation (2 years postbaseline) there was evidence of increased neurocognitive impairment suggestive of greater dysfunction of the patient's right, versus left, cerebral hemisphere. Overall, the patient's neuropsychological profile coincided with the Syndrome of Nonverbal Learning Disabilities as proposed by Rourke and his colleagues (Rourke, 1987, 1988, 1995; Rourke & Tsatsanis, 1996). These findings are discussed in light of the Syndrome of Nonverbal Learning Disabilities (and the related white matter model) and the possible negative impact of the patient's pineal tumor and subsequent chemotherapy and cranio-spinal irradiation on her neuropsychological functioning. PMID- 10379417 TI - Affective verbal memory in patients with temporal lobe epilepsy. AB - Eighteen epileptic patients with unilateral temporal lobe epilepsy (9 left, 9 right) were evaluated with a verbal memory task involving recall of 2 stories, 1 with affective content and 1 that was neutral. A trend for better performance by the group with intact left hemispheres was found for a quantitative score of number of story units recalled. For a qualitative score of number of symbolic distortions, a main effect of affective load was found, such that more distortions were made for the story with affective than neutral content. This effect remained significant when the left temporal lobe epilepsy patients were analyzed separately and was not found for the right temporal lobe epilepsy patients alone. Additional analyses for the subset of 5 patients with left and 6 patients with right temporal lobectomies involving removal of the hippocampus and amygdala were in the same direction as the analyses for all 18 participants. These findings are consistent with other reports of material-specific memory deficits, such that verbal memory deficits are associated with left temporal lobe epilepsy. The differences between performance on the affective and neutral stories for the left and right temporal lobe epilepsy patients are discussed and related to the role of the amygdala in affective processing. PMID- 10379418 TI - Paper or plastic: another ecological consideration in neuropsychological assessment. AB - The high volume of paper involved in the generation of neuropsychological evaluations has implications for the environment, for the cost of providing services, and for the space required to store records. One solution is to insert single-use test forms into plastic sheet protectors and provide the test taker with a fine-tipped washable marker. This study investigated whether this modification in test administration practice affects performance. Comparisons were made for 6 different neuropsychological tests. No significant differences in performance were found when comparing standard administration practice to this more ecologically minded alternative. PMID- 10379419 TI - Metabolism of some "second"- and "fourth"-generation antidepressants: iprindole, viloxazine, bupropion, mianserin, maprotiline, trazodone, nefazodone, and venlafaxine. AB - 1. This review summarizes the major known aspects of the metabolism of second generation (iprindole, viloxazine, bupropion, mianserin, maprotiline, and trazodone) and fourth-generation (nefazodone and venlafaxine) antidepressants. 2. Discussions about specific enzymes involved and about possible pharmacokinetic drug-drug interactions, particularly as they relate to cytochrome P450 enzymes, are provided. PMID- 10379420 TI - Metabolism and pharmacokinetics of selective serotonin reuptake inhibitors. AB - 1. Five drugs with the predominant pharmacologic effect of inhibiting the neuronal reuptake of serotonin are available worldwide for clinical use. This class of psychoactive drugs, known as selective serotonin reuptake inhibitors (SSRIs), is comprised of fluoxetine, sertraline, paroxetine, fluvoxamine, and citalopram. 2. The SSRIs appear to share similar pharmacodynamic properties which translate to efficacy in the treatment of depression and anxiety syndromes. The drugs are differentiated by their pharmacokinetic properties with regard to stereochemistry, metabolism, inhibition of cytochrome enzymes, and participation in drug-drug interactions. Studies focusing on the relationship of plasma drug concentration to therapeutic and adverse effects have not confirmed the value of plasma concentration monitoring. 3. This review summarizes the metabolism and relevant pharmacokinetic properties of the SSRIs. PMID- 10379421 TI - Comparison of the effects of antidepressants and their metabolites on reuptake of biogenic amines and on receptor binding. AB - 1. The present survey compares the effects of antidepressants and their principal metabolites on reuptake of biogenic amines and on receptor binding. The following antide-pressants were included in the study: the tricyclic antidepressants amitriptyline, dothiepin, and lofepramine and the atypical antidepressant bupropion, which all have considerable market shares in the UK and/or US markets; the selective serotonin reuptake inhibitors (SSRIs) citalopram, fluoxetine, fluvoxamine, paroxetine, and sertraline; and the recently approved antidepressants venlafaxine and nefazodone. 2. Amitriptyline has similar in vitro reuptake inhibitory potencies for 5-HT and NA, whereas the metabolite nortriptyline is preferentially a NA reuptake inhibitor. Both amitriptyline and nortriptyline are also 5-HT2 receptor antagonists. 3. Dothiepin has equipotent 5 HT and NA reuptake inhibitory activity, whereas northiaden shows a slight selectivity for NA reuptake inhibition. Dothiepin and northiaden are also 5-HT2 receptor antagonists. The slow elimination rate of northiaden (36-46 hr) compared to dothiepin (14-24 hr) suggests that northiaden contributes significantly to the therapeutic effect of dothiepin. 4. Lofepramine is extensively metabolized to desipramine. Desipramine plays an important role in the antidepressant activity of lofepramine, as the plasma elimination half-life of lofepramine (4-6 hr) is much shorter than that of desipramine (24 hr). Both compounds are potent and selective inhibitors of NA reuptake. 5. The five approved SSRIs, citalopram, fluoxetine, fluvoxamine, paroxetine, and sertraline, are potent 5-HT reuptake inhibitors, and the demethyl metabolites, norfluoxetine, demethylsertraline, and demethylcitalopram, also show selectivity. Paroxetine and sertraline are the most potent inhibitors of 5-HT reuptake, whereas citalopram is the most selective. Fluoxetine is the least selective and the metabolite of fluoxetine, norfluoxetine, is a more selective and more potent 5-HT reuptake inhibitor than the parent compound and has an extremely long half-life (7-15 compared to 1-3 days). Thus the metabolite plays an important role for the therapeutic effect of fluoxetine. Fluoxetine is also a 5-HT2C receptor antagonist. Demethylsertraline is a weaker and less selective 5-HT reuptake inhibitor in vitro than sertraline, but demethylsertraline has a very long half-life (62-104 hr) compared to the parent compound (24 hr) and it might play a role in the therapeutic effects of sertraline. Demethylcitalopram has about a 10 times lower 5-HT reuptake inhibitory potency in vitro than citalopram, and the elimination half-lives are approximately 1.5 and 2 days, respectively. 6. Bupropion and hydroxybupropion are weak inhibitors of biogenic amine reuptake. The mechanisms of action responsible for the clinical effects of bupropion are not fully understood, but it has been suggested that both dopaminergic and noradrenergic components play a role and that the hydroxybupropion metabolite contributes significantly to the antidepressant activity. 7. Venlafaxine and O-demethylvenlafaxine are weak inhibitors of 5-HT and NA reuptake, and the selectivity ratios are close to one. O-Demethylvenlafaxine is eliminated more slowly than venlafaxine (plasma half lives of 5 and 11 hr, respectively), and it is likely that it contributes to the overall therapeutic effect of venlafaxin. 8. Nefazodone and alpha hydroxynefazodone are equipotent 5-HT and NA reuptake inhibitors. Both compounds are also 5-HT2 receptor antagonists. Both parent compound and metabolite have short elimination half-lives. PMID- 10379425 TI - [Quality assurance of infection-prevention in surgical procedures]. PMID- 10379424 TI - Metabolism of anxiolytics and hypnotics: benzodiazepines, buspirone, zoplicone, and zolpidem. AB - 1. The benzodiazepines are among the most frequently prescribed of all drugs and have been used for their anxiolytic, anticonvulsant, and sedative/hypnotic properties. Since absorption rates, volumes of distribution, and elimination rates differ greatly among the benzodiazepine derivatives, each benzodiazepine has a unique plasma concentration curve. Although the time to peak plasma levels provides a rough guide, it is not equivalent to the time to clinical onset of effect. The importance of alpha and beta half-lives in the actions of benzodiazepines is discussed. 2. The role of cytochrome P450 isozymes in the metabolism of benzodiazepines and in potential pharmacokinetic interactions between the benzodiazepines and other coadministered drugs is discussed. 3. Buspirone, an anxiolytic with minimal sedative effects, undergoes extensive metabolism, with hydroxylation and dealkylation being the major pathways. Pharmacokinetic interactions of buspirone with other coadministered drugs seem to be minimal. 4. Zopiclone and zolpidem are used primarily as hypnotics. Both are extensively metabolized; N-demethylation, N-oxidation, and decarboxylation of zopiclone occur, and zolpidem undergoes oxidation of methyl groups and hydroxylation of a position on the imidazolepyridine ring system. Zopiclone has a chiral centre, and demonstrates stereoselective pharmacokinetics. Metabolic drug drug interactions have been reported with zopiclone and erythromycin, trimipramine, and carbamazepine. Reports to date indicate minimal interactions of zolpidem with coadministered drugs; however, it has been reported to affect the Cmax and clearance of chlorpromazepine and to decrease metabolism of the antiviral agent ritonavin. Since CYP3A4 has been reported to play an important role in metabolism of zolpidem, possible interactions with drugs which are substrates and/or inhibitors of that CYP isozyme should be considered. PMID- 10379422 TI - Metabolism, pharmacogenetics, and metabolic drug-drug interactions of antipsychotic drugs. AB - 1. Antipsychotic drugs are extensively metabolised by cytochrome P450 (CYP) enzymes. 2. Dispositions of a number of antipsychotic drugs have been shown to cosegregate with polymorphism of CYP2D6. 3. Metabolic drug-drug interactions have frequently been observed when antipsychotics are coadministered with other drugs. 4. Many antipsychotic drugs are converted to active metabolites which can contribute to the therapeutic or side effects of the parent drug. 5. Information concerning the individual CYP isoenzymes involved in the metabolism of antipsychotic drugs is important for the safe clinical use of this group of drugs. PMID- 10379426 TI - Medicaid eligibility expansion in Florida: effects on maternity care financing and the delivery system. AB - CONTEXT: In July 1989, the income limit on Medicaid eligibility for pregnant women in Florida was increased from 100% to 150% of the poverty level. This change may have led to substantial shifts in the financing of pregnancy-related care, and also may have had distinct effects on different providers in the health care delivery system. METHODS: Matched birth and death certificates, hospital discharge abstracts, Medicaid eligibility records and encounter records from county public health departments were used to estimate changes in the flows of funds and services by major payer groups during the period preceding the expansion (July 1988-June 1989) and for calendar year 1991. A total of 188,793 births in the first period and 193,292 in the second were examined. RESULTS: The number of births financed annually by Medicaid in Florida increased by 47% following the eligibility expansion, from 47,400 in 1988-1989 to 69,600 in 1991. This increase stemmed largely from covered births to women who otherwise would have been uninsured. Seventy-three percent of the additional 22,200 deliveries funded through Medicaid in 1991 are attributed to women who were eligible as a result of the expansions. The additional prenatal care financed by Medicaid was delivered almost entirely by county public health departments, which increased their capacity by more than 100%, from 177,000 visits in 1988-1989 to 433,000 in 1991. Medicaid payments for maternity care increased 39%, from $135 million to $187 million, while payments made by the uninsured dropped by 29%. These changes resulted in a 5% rise in hospital revenues, despite little change in the number of admissions. CONCLUSIONS: The Medicaid expansion benefited low-income pregnant women and hospitals in Florida. It is unknown whether the private delivery system would have accommodated the increased demand in the absence of the public health system response. PMID- 10379423 TI - Metabolism and excretion of mood stabilizers and new anticonvulsants. AB - 1. The mood stabilizers lithium, carbamazepine (CBZ), and valproate (VPA), have differing pharmacokinetics, structures, mechanisms of action, efficacy spectra, and adverse effects. Lithium has a low therapeutic index and is renally excreted and hence has renally-mediated but not hepatically-mediated drug-drug interactions. 2. CBZ has multiple problematic drug-drug interactions due to its low therapeutic index, metabolism primarily by a single isoform (CYP3A3/4), active epoxide metabolite, susceptibility to CYP3A3/4 or epoxide hydrolase inhibitors, and ability to induce drug metabolism (via both cytochrome P450 oxidation and conjugation). In contrast, VPA has less prominent neurotoxicity and three principal metabolic pathways, rendering it less susceptible to toxicity due to inhibition of its metabolism. However, VPA can increase plasma concentrations of some drugs by inhibiting metabolism and increase free fractions of certain medications by displacing them from plasma proteins. 3. Older anticonvulsants such as phenobarbital and phenytoin induce hepatic metabolism, may produce toxicity due to inhibition of their metabolism, and have not gained general acceptance in the treatment of primary psychiatric disorders. 4. The newer anticonvulsants felbamate, lamotrigine, topiramate, and tiagabine have different hepatically-mediated drug-drug interactions, while the renally excreted gabapentin lacks hepatic drug-drug interactions but may have reduced bioavailability at higher doses. 5. Investigational anticonvulsants such as oxcarbazepine, vigabatrin, and zonisamide appear to have improved pharmacokinetic profiles compared to older agents. 6. Thus, several of the newer anticonvulsants lack the problematic drug-drug interactions seen with older agents, and some may even (based on their mechanisms of action and preliminary preclinical and clinical data) ultimately prove to have novel psychotropic effects. PMID- 10379427 TI - Teenage partners' communication about sexual risk and condom use: the importance of parent-teenager discussions. AB - CONTEXT: Teenagers' communication with their partners about sex and their use of condoms may be influenced by the discussions teenagers have with their parents about sex. However, little is known about the process of parent-teenager communication on this topic. Understanding both what parents discuss with their children and how they discuss it may lead to a greater understanding of teenagers' sexual behavior. METHODS: Interviews were conducted with 372 sexually active black and Hispanic youth aged 14-17 from Alabama, New York and Puerto Rico. Regression analyses were used to examine parent-teenager discussions about sexuality and about sexual risk, and parental communication skills as predictors of teenagers' discussions about sexual risk with a partner and teenagers' condom use. RESULTS: Parent-teenager discussions about sexuality and sexual risk were associated with an increased likelihood of teenager-partner discussions about sexual risk and of teenagers' condom use, but only if parents were open, skilled and comfortable in having those discussions. Teenagers' communication with their partner about sexual risk also was associated with greater condom use, but the relationship between parent-teenager communication and teenagers' condom use was independent of this association. CONCLUSIONS: The influence on teenagers of parent-teenager discussions about sexuality and sexual risk depends on both what parents say and how they say it. Programs that foster parent-teenager communication about sexuality and sexual risk must emphasize both of these aspects. PMID- 10379428 TI - Pregnancy rates among U.S. women and their partners in 1994. AB - CONTEXT: When rates of pregnancy, birth and abortion are calculated only for the women involved, men's role in reproduction is ignored, resulting in limited understanding of their influence on these outcomes. METHODS: Data from the 1995 National Survey of Family Growth and from the 1994-1995 Alan Guttmacher Institute Abortion Patient Survey were combined with national natality statistics to estimate pregnancy rates in 1994 for women and their male partners, by age and marital status at the time of conception. RESULTS: Nine percent of both men and women aged 15-44 were involved in conceiving a pregnancy in 1994 (excluding those resulting in miscarriages). Pregnancy levels were highest among women aged 20-24 and among male partners aged 25-29. Men younger than 20 were involved in about half as many pregnancies as were women this age (9% compared with 18%). In contrast, men aged 35 and older were involved in roughly twice as many pregnancies as were similarly aged women (19% compared with 9%). Three out of every four pregnancies in 1994 resulted in a birth. However, 47% of pregnancies involving men younger than 18 ended in abortion, compared with about 34% of those involving men aged 40 and older. In comparison, 31% of pregnancies among women younger than 18 resulted in abortion, while 39% of those among women aged 40 and older were terminated. CONCLUSION: The overall rate at which men were involved in causing a pregnancy is similar to the pregnancy rate among women. Men are typically older than women when they are involved in a pregnancy, however. This implies that men may bring more experience and resources to the pregnancy experience. PMID- 10379429 TI - Sexual intercourse, abuse and pregnancy among adolescent women: does sexual orientation make a difference? AB - CONTEXT: Although a limited amount of research has retrospectively explored the childhood and adolescent heterosexual experiences of lesbians, little is known about the prevalence of heterosexual behavior and related risk factors or about pregnancy histories among lesbian and bisexual teenagers. METHODS: A secondary analysis was conducted using responses from a subsample of 3,816 students who completed the 1987 Minnesota Adolescent Health Survey. Behaviors, risk factors and pregnancy histories were compared among adolescents who identified themselves as lesbian or bisexual, as unsure of their sexual orientation and as heterosexual. RESULTS: Overall, bisexual or lesbian respondents were about as likely as heterosexual women ever to have had intercourse (33% and 29%, respectively), but they had a significantly higher prevalence of pregnancy (12%) and physical or sexual abuse (19-22%) than heterosexual or unsure adolescents. Among sexually experienced respondents, bisexual or lesbian and heterosexual women reported greater use of ineffective contraceptives (12-15% of those who used a method) than unsure adolescents (9%); bisexual or lesbian respondents were the most likely to have frequent intercourse (22%, compared with 15-17% of the other groups). In the sample overall, among those who were sexually experienced and among those who had ever been pregnant, bisexual or lesbian women were the most likely to have engaged in prostitution during the previous year. CONCLUSIONS: Providers of reproductive health care and family planning services should not assume that pregnant teenagers are heterosexual or that adolescents who say they are bisexual, lesbian or unsure of their sexual orientation are not in need of family planning counseling. Further research should explore the interactions between adolescent sexual identity development and sexual risk behaviors. PMID- 10379430 TI - Live births resulting from unintended pregnancies: is there variation among states? The PRAMS Working Group. AB - CONTEXT: States need data on live births resulting from unintended pregnancies in order to assess the need for family planning services; however, many states do not collect such data. Some states may use extrapolated rates from other states. METHODS: Pregnancy Risk Assessment Monitoring System (PRAMS) data were assessed to explore the feasibility of extrapolating data on the percentage of live births resulting from unintended pregnancies from states that collect these data to states that do not. Data on women who had live births between 1993 and 1995 were examined for eight states: Alabama, Florida, Georgia, Michigan, New York (excluding New York City), Oklahoma, South Carolina and West Virginia. Logistic regression was used to determine state variation in the odds of delivering a live birth resulting from an unintended pregnancy after adjustment for maternal race, marital status, age, education, previous live birth and participation in the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC). RESULTS: The percentage of live births resulting from unintended pregnancy ranged from 33% in New York to 49% in Alabama, Georgia and South Carolina. Compared with women in Alabama, women in Oklahoma were more likely to deliver a live birth resulting from an unintended pregnancy (odds ratio of 1.2, confidence interval of 1.1-1.3) and women in New York State were less likely (odds ratio of 0.7, confidence interval of 0.6-0.8) to have such a birth. However, unmarried white women in New York had lower odds of having a live birth resulting from an unintended pregnancy and married black women in Michigan had higher odds of having a live birth resulting from unintended pregnancy than their counterparts in Alabama. Although the percentages varied, in all eight states women who were black, were unmarried, were younger than 20 years of age, had less than 12 years of education or had more than one child had higher percentages of live births resulting from unintended pregnancy than women with other demographic characteristics. CONCLUSIONS: Data on which women have the greatest risk of delivering a live birth resulting from an unintended pregnancy may be extrapolated from one state to another, but the rate of such births may overestimate or underestimate the problem from one state to another. PMID- 10379431 TI - Psychosocial and educational services for female college students with genital human papillomavirus infection. AB - CONTEXT: College-age women have a high risk of acquiring human papillomavirus (HPV) infection, which may have substantial psychosocial and physical effects. Young women who become infected need information and support from health care professionals, but little is known about providers' attitudes toward or provision of interventions for helping women cope with HPV. METHODS: A survey of 73 nurse practitioners and 70 physicians in college-based health clinics explored their perceptions of the need for psychosocial and educational interventions and their practices regarding such services for HPV patients. Analysis of variance and chi square testing were used to examine differences by providers' type and gender. RESULTS: At least 86% of providers agree that HPV infection has a variety of psychosocial effects on young women, but only 54% spend at least 10 minutes providing education and counseling to all of their HPV patients. Roughly 80-90% routinely take a sexual history, explain the potential of HPV recurrence and discuss the risk of cancer with HPV patients; however, fewer than half always offer a variety of other interventions that could help patients cope with the diagnosis and promote preventive behaviors. Female providers are more aware of the psychosocial impact of HPV and the need for support than are male providers. However, nurse practitioners provide counseling and educational interventions more frequently than do physicians, even when gender is controlled for. CONCLUSIONS: College-based health providers need to improve the content of the counseling and education they offer to women with HPV, as well as the consistency with which they deliver those interventions. When they are unable to provide services, they should be able to refer patients elsewhere. PMID- 10379432 TI - The state bonus to reward a decrease in 'illegitimacy': flawed methods and questionable effects. PMID- 10379433 TI - The 'new Medicaid': an incremental path to national health care reform. PMID- 10379434 TI - Reasons for infecundity. PMID- 10379435 TI - Catheter-associated primary bloodstream infections: epidemiology and preventive methods. AB - The incidence of catheter-associated primary bloodstream infections (BSIs) in Germany as defined by the CDC (Centers for Disease Control) has been determined on the basis of (a) a national prevalence study in a representative sample of 72 hospitals (NIDEP), and (b) an incidence study in which data about the use and duration of insertion of central venous catheters (CVC) and of catheter associated BSI were collected from 25 intensive-care units (ICUs) participating in the hospital infection surveillance system (KISS+) and analyzed. The first study showed primary bloodstream infection to be the fourth most frequent nosocomial infection at 8.3% of all nosocomial infections. With an ICU prevalence of 2.1%, primary BSI comprises 12.8% of all nosocomial infections observed in ICU patients. The second study showed a 60.4% prevalence of CVC use in German ICUs. An analysis of 55,400 CVC days in 14,988 ICU patients in the KISS hospitals yielded 2.2 CVC-associated BSIs per 1,000 CVC days (CI95 1.8-2.6). The rates of CVC-associated BSI on individual hospital wards were very variable and indicates a reduction potential. A reduction in the number of infections of about one-third would prevent 1,000-1,400 deaths due to CVC-associated BSI annually as well as about 40,000 to 60,000 extra days of hospital stay and the associated costs. PMID- 10379436 TI - Basic aspects of the pathogenesis of staphylococcal polymer-associated infections. PMID- 10379437 TI - Prevention of infections caused by central venous catheters--established and novel measures. AB - Intravascular catheters play an important role in infections in intensive care and hemodialysis patients. This becomes evident only if full microbiological diagnoses are made. Difficulties in the diagnosis and treatment of microbially colonized catheters make the prevention of infection particularly important. The most important preventive measures are a strict evaluation of the indications for the use of the catheter and strict hygienic precautions during insertion and maintenance of the central venous catheter. Other measures, some of which are controversial, may be considered, such as the specific decontamination of Staphylococcus carriers using mupirocin. A new approach in the prevention of catheter-related infections is the use of catheter materials impregnated with antibiotics, antiseptics or metals. Slow-delivery systems release the antimicrobially active substance from the catheter material and thus reduce the proliferation of adherent bacteria. Some of these slow-delivery systems have been used in clinical trials, with varied results. Current research is directed towards the prevention of the first stage in the pathogenesis of catheter associated infections, namely the adherence of bacteria to the catheter polymer, e.g. by impregnation of the polymer with silver. Laboratory studies, animal experiments and initial clinical trials suggest that it will soon be possible to reduce the frequency of catheter-associated infections to below the levels attainable with current general and specific preventive measures, through the use of coated catheters. PMID- 10379438 TI - A new technology of microdispersed silver in polyurethane induces antimicrobial activity in central venous catheters. AB - Metal ions or metal ions in complexes or compounds have been used for centuries to disinfect fluids, solids and tissues. The biocidal effect of silver, with its broad spectrum of activity including bacterial, fungal and viral agents, is particularly well known and the term "oligodynamic activity" was coined for this phenomenon. Silver ions have an affinity to sulfhydryl groups in enzyme systems of the cell wall, through which they interfere with the transmembranous energy transfer and electron transport of bacterial microorganisms. Silver ions also block the respiratory chain of microorganisms reversibly in low concentrations and irreversibly in higher concentrations. Binding to the DNA of bacteria and fungi increases the stability of the bacterial double helix and thus inhibits proliferation. There is no cross resistance with antibiotics and also no induction of antimicrobial resistance by silver ions. The concentrations required for bactericidal activity are in the range 10(-9) mol/l. These concentrations can be achieved in solution by the interaction of metallic silver with electrolytes only if there is a large enough surface of silver. By a novel technology, metallic silver is distributed in submicron particles in polyurethane and results in a concentration of 0.8% in an active surface of 450 cm2/g polyurethane. Polyurethane is hygroscopic and rapidly attracts water; the interaction of electrolyte solutions with the extremely finely distributed silver throughout the polyurethane releases bactericidal concentrations of silver ions over a period of years to the surface of the material. The electronegatively charged surface of bacteria attracts the positively charged silver ions. The concentrations released from the polyurethane are far below the toxic concentrations for humans. PMID- 10379439 TI - The Erlanger silver catheter: in vitro results for antimicrobial activity. AB - The antimicrobial activity of a silver-impregnated polymer catheter (the Erlanger silver catheter) was demonstrated by determining the microbial adhesion to the surface of the catheter and by measuring the rate of proliferation (viability) of microorganisms at this site. On the surface of a catheter impregnated with silver, according to previously described methods, the bacterial adhesion of Staphylococcus epidermidis is reduced by 28-40%. Bacterial proliferation on the surface of the catheter and biofilm production are also substantially reduced by the elution of free silver ions from the catheter matrix. Bacteriostatic and bactericidal activities can be determined. The antimicrobial efficacy of the silver catheter is not reduced by blood components. There is no loss in antimicrobial activity for weeks after preincubation in water or phosphate buffered saline. The antimicrobial activity depends on the extent of the active silver surface. PMID- 10379440 TI - Thrombogenicity testing of central venous catheters in vitro. AB - To date there have been no standard methods for assessing the thrombogenicity of central venous catheters. A procedure for testing the thrombogenicity of intravenous lines such as the silver-impregnated catheter by continuous blood flow in vitro was therefore developed. For this test, fresh blood was drawn from healthy human donors and anti-coagulated with sodium citrate (1:9). All material tested (catheter tubes with and without silver manufactured in the same way, polyethylene tubes and tubes with potentially thrombogenic material) were perfused through their lumen with anticoagulated blood for up to 31 hours. Blood samples were collected at different times from the test system at sites before and after the perfusion of the test catheters. The hemoglobin concentration, erythrocyte, leukocyte and thrombocyte counts and markers for thrombin activation (thrombin-antithrombin III-complex, F1 + 2)-prothrombin fragments) and for hyperfibrinolysis (d-dimers) were determined. No thrombin activation or signs of hyperfibrinolysis were detected in any material tested. Polyethylene tubes were found to cause hemolysis, as shown by a decrease in hemoglobin content from 15 g% to 4.5 g%. Tecothane tubes with and without silver did not induce hemolysis. PMID- 10379441 TI - Analysis of the acute cytotoxicity of the Erlanger silver catheter. AB - The Erlanger silver catheter consists of a new form of polyurethane, which contains finely dispersed metallic silver. The aim of this study was to establish the biocompatibility of this intravenous catheter by investigating the acute cytotoxicity of extracts from the Erlanger silver catheter on human fibroblasts and lymphocytes. Extracts of the Erlanger silver catheter were not cytotoxic for MRC-5 human fibroblasts nor for sensitized phytohemagglutinin (PHA)-stimulated human lymphocytes. The addition of silver powder of up to 2% by weight to the basic catheter polyurethane Tecothane led to no increase in acute cytotoxicity in comparison with untreated Tecothane. The Erlanger silver catheter is a new intravenous catheter with good biocompatibility. PMID- 10379442 TI - Biocompatibility testing of a new silver-impregnated catheter in vivo. AB - The purpose of this investigation was to compare the local effects of polyurethane (Tecothane) and silicone tubes with or without silver impregnation in rats. Bacterial colonization or infection of the exit site and/or tunnel were documented and interpreted. All tubes were placed subcutaneously or percutaneously in the neck of 41 Sprague-Dawley rats and guided beneath the dorsal muscles into the peritoneal cavity. The incidence of bacterial abscesses along the implanted tubes was evaluated daily. After 90 days, or earlier if sepsis developed, the animals were killed painlessly and various organs and tissues from the entry site and the catheter tunnel examined histologically. In the group where polyurethane tubes were placed percutaneously, there was no difference in the frequency of abscesses between silver-impregnated and non impregnated tubes (5/6 with and 5/7 without silver). The only difference noted was in the group with percutaneously placed silicone tubes between those with and without silver. Abscesses only occurred in 2/4 animals in the silver group and in 5/5 animals in the control group. Histological examination showed no difference in either group between infectious and foreign body reactions. Silver particles in subcutaneous, muscle and peritoneal tissue could not be demonstrated. PMID- 10379443 TI - Thermoplastic polyurethane--the material used for the Erlanger silver catheter. AB - The Erlanger silver catheter consists of a thermoplastic polyurethane elastomer (TPU). To achieve an antimicrobial effect, silver particles at a concentration of 0.5-1.0% by weight are incorporated into the basic material. The good mechanical properties were shown to remain present after the incorporation of the silver. The characteristic mechanical data of blends made of a soft and a hard TPU type approximated closely to a linear relationship with the ratio of the mixture. The buckling stability of catheters as a function of their cross section could be described by Euler's formula. This enables the buckling stability to be calculated when material constants and catheter geometry are known. PMID- 10379444 TI - Determination of the silver ion release from polyurethanes enriched with silver. AB - The Erlanger silver catheter was developed in order to reduce the risk of infection from long-term catheters by means of silver ions, which are known to have antibacterial properties. This is achieved by incorporating silver into polyurethane catheters by means of a special procedure. The aim of this materials science study was to verify the release of silver ions from the polyurethanes. Static experiments were carried out following the usual norms. Clinically relevant dynamic experiments, which were designed and constructed at this institute, were also performed. The eluates from both experiments were analyzed by anodic stripping voltammetry. Polyurethanes filled with silver, as used in the Erlanger silver catheter, release silver in static as well as in dynamic experiments. If the experimentally determined releases are converted to the usual catheter length of 30 cm, the release is about 0.1 microgram/l. This lies in the order of concentrations that have been reported in the literature to be antibacterial. PMID- 10379445 TI - A new scoring system for the clinical diagnosis of catheter-related infections. AB - It is difficult to make the clinical diagnosis of catheter-related infections using the available and established definitions of the HICPAC (Hospital Infection Control Practices Advisory Committee) of the CDC (Centers for Disease Control, definitions of nosocomial infections). The scoring system shown here is a modification of these definitions and has enabled the causal relationship between the catheter and clinical episodes of systemic infections to be quantitatively graded. The scoring system included the following criteria: height and rate of rise of body temperature, attendant shivering, identification of pathogens in blood and/or catheter tip cultures, improvement in the clinical course after catheter removal, signs of catheter exit site inflammation and results of diagnostic tests for other possible sources of infection. These criteria were graded using points and weighted according to their specificity. The comparative evaluation of 65 episodes of systemic infections using the scoring system and the diagnostic criteria of HICPAC showed agreement in 85%. No case was graded "false negative." In nine of ten false-positive cases additional findings supported the presence of a catheter-associated infection. This scoring system appears, therefore, to be more sensitive than existing diagnostic criteria, without loss of specificity. PMID- 10379446 TI - Silver catheter study: methods and results of microbiological investigations. AB - Within the framework of the clinical study of the Erlangen silver catheter 104 silver catheters and 105 control catheters were tested by microbiological culture. This was done by rolling the catheter on a blood agar plate, washing the lumen through with tryptic soy broth (TSB) and, after ultrasound treatment, incubating the catheter tip in TSB as an enrichment culture for detecting very low bacterial counts. There was good agreement in the numbers of colony-forming units (CFU) detected by the roll plate and luminal washout cultures in 92% of the silver and 89% of the control catheters tested. Seventy-six (73%) of the 104 silver catheters showed no bacterial growth and 16 (15%) showed very low bacteria counts (< 15 CFU), or growth only after enrichment, which were attributed in both instances to catheter contamination. Twelve catheter tips (12%) showed significant bacterial counts greater than 15 CFU which were indicative of colonization or catheter-related infection. Corresponding results in the control catheters were 59 (56%), 28 (27%) and 18 (17%), respectively, a higher rate of infection or contamination which was statistically significant (chi-square test: P = 0.04). PMID- 10379448 TI - Clinical study of the Erlanger silver catheter--data management and biometry. AB - The clinical evaluation of venous catheters for catheter-induced infections must conform to a strict biometric methodology. The statistical planning of the study (target population, design, degree of blinding), data management (database design, definition of variables, coding), quality assurance (data inspection at several levels) and the biometric evaluation of the Erlanger silver catheter project are described. The three-step data flow included: 1) primary data from the hospital, 2) relational database, 3) files accessible for statistical evaluation. Two different statistical models were compared: analyzing the first catheter only of a patient in the analysis (independent data) and analyzing several catheters from the same patient (dependent data) by means of the generalized estimating equations (GEE) method. The main result of the study was based on the comparison of both statistical models. PMID- 10379447 TI - Reduced rates of catheter-associated infection by use of a new silver-impregnated central venous catheter. AB - A central venous catheter with a new form of silver impregnation of the internal and external surfaces was investigated for antimicrobial activity and tolerance in patients in a controlled comparative, prospective and randomized clinical study. Commercially available catheters with no antimicrobial activity were used as controls. One hundred sixty-five catheters were included in the final evaluation. All catheters were percutaneously inserted for the first time with a duration of > or = 5 days and a microbiological examination of the catheter tip. Catheter location (> 90% internal jugular vein), mean duration of catheterization (8-9 days), patients' age and diagnosis were comparable in both groups. Silver impregnated catheter tips showed an incidence of colonization in 14.2/1000 catheter days and control catheters in 22.8/1000 catheter days. This represents a reduction of 37.7%. Catheter-associated infections were diagnosed in the silver group in 5.26/1000 catheter days and 18.34/1000 catheter days in the control group, indicating a reduction rate of 71.3% (P < 0.05, chi 2-test). No complications or side effects were documented in either group. PMID- 10379449 TI - Reducing catheter-associated infections with silver-impregnated catheters in long term therapy of children. AB - Central venous long-term catheters offer reliable, large-lumen vascular access with high flow rates for delivery of nutrition or for cell-containing infusions and perfusions. Catheter-associated infections (CAI) pose the greatest threat to such vascular access, despite existing preventive measures. In this article one prospective and one retrospective study of CAI in pediatric therapy are presented. Study I: A retrospective investigation from 1990 through 1995 of 60 conventional long-term catheters in 50 patients. The total number of days in which the catheters were in place was 11,818. The calculated CAI incidence was 1 per 1,000 days of catheter insertion. Bacteriologically demonstrated CAI (identical isolate on the catheter tip and in a blood culture) occurred in three instances (5%). Five cases (8.3%) were diagnosed with a therapy-resistant, septic clinical picture. Study II: A prospective, randomized comparison of long-term silver-impregnated (Erlanger silver catheters) and control catheters (Quinton Instrument Co.) was made with 41 patients (20 with a silver catheter, 21 with a Quinton catheter). To date, the silver catheters have been distinguished by sterile bacteriological findings, whereas three cases of CAI have been demonstrated with the comparative catheters. One patient recently underwent intensive care after becoming unstable with signs of septic shock and demonstrable Pseudomonas aeruginosa, and two other patients manifested coagulase negative staphylococci on the catheter tips. In three of nine control catheters an incidence of 1.18 per 1,000 days of indwelling catheters was found, whereas no CAI has occurred with the eight microbiologically tested silver catheters. PMID- 10379450 TI - Catheter-associated infections in urology: possible use of silver-impregnated catheters and the Erlanger silver catheter. AB - Indwelling urinary catheters play a very important part in urology. However, their use is accompanied by a considerable increase in the risk of nosocomial urinary tract infections. The pathophysiological cause is ascribed to pathogens that adhere to the catheter surface, proliferate and produce a biofilm. In addition to aseptic techniques, modification of the catheter material to confer antimicrobial activity plays an essential part in the prevention of catheter related urinary tract infections. The antimicrobial efficacy of silver against gram-positive and gram-negative bacteria is well known and amply shown in vitro. The efficacy of silver-impregnated catheters is critically dependent on both the chemical structure of the incorporated silver and the way the silver has been combined with the basic catheter material. Hence, clinical studies on silver modified catheters have so far given inconsistent results. The new technology of the Erlanger silver catheter offers the opportunity of an effective reduction in catheter-related infections. PMID- 10379451 TI - Economic aspects of innovations in medical technology. PMID- 10379452 TI - Health and equity in the world in the era of "globalization". AB - The author critically discusses some of the major arguments given for the growth of inequalities in health in the world today. He also questions the "technocratic," "humanistic," or "apolitical" discourse used by most international agencies in their analysis of the growing inequalities, a discourse that obscures the actual causes of this growth: the power relations among and within countries. PMID- 10379453 TI - A fundamental shift in the approach to international health by WHO, UNICEF, and the World Bank: instances of the practice of "intellectual fascism" and totalitarianism in some Asian countries. AB - Navarro has used the term "intellectual fascism" to depict the intellectual situation in the McCarthy era. Intellectual fascism is now more malignant in the poor countries of the world. The Indian Subcontinent, China, and some other Asian countries provide the context. The struggles of the working class culminated in the Alma-Ata Declaration of self-reliance in health by the peoples of the world. To protect their commercial and political interests, retribution from the rich countries was sharp and swift, they "invented" Selective Primary Health Care and used WHO, UNICEF, the World Bank, and other agencies to let loose on poor countries a barrage of "international initiatives" as global programs on immunization, AIDS, and tuberculosis. These programs were astonishingly defective in concept, design, and implementation. The agencies refused to take note of such criticisms when they were published by others. They have been fascistic, ahistorical, grossly unscientific, and Goebbelsian propagandists. The conscience keepers of public health have mostly kept quiet. PMID- 10379454 TI - Toward an ecosocial view of health. AB - The changing patterns of health in the United States justify both celebration and dismay. We can celebrate declining mortality rates, increased life expectancy, and improvements in diagnostic and therapeutic technologies. But public health was caught by surprise by the return of infectious disease; the gap in health outcomes between rich and poor and between whites and blacks increases; there is a growing discrepancy between what is technically possible and the actual health status; and despite its greater expenditures on health, the United States lags behind the other developed countries in health outcomes. The authors examine four reasons for this: we do not buy more health care, only pay more for it; we receive more health care, but much of it inappropriate, ineffective, or harmful; only some of us get more health care; and we have created a way of life that makes us sick, then spend more to repair the damage. Major failures arise when problems are understood too narrowly. An ecosocial perspective attempts to look at the whole. It rejects as false the dichotomies social/biological, physical/psychological, genetic/environmental, lifestyle/environment, examining their interrelations rather than assigning them relative weights. In addition to looking at average differences among populations, the authors examine patterns of variability in health outcomes. PMID- 10379455 TI - Embodying inequality: a review of concepts, measures, and methods for studying health consequences of discrimination. AB - Investigating effects of discrimination upon health requires clear concepts, methods, and measures. At issue are both economic consequences of discrimination and accumulated insults arising from everyday and at times violent experiences of being treated as a second-class citizen, at each and every economic level. Guidelines for epidemiologic investigations and other public health research on ways people embody racism, sexism, and other forms of social inequality, however, are not well defined, as research in this area is in its infancy. Employing an ecosocial framework, this article accordingly reviews definitions and patterns of discrimination within the United States; evaluates analytic strategies and instruments researchers have developed to study health effects of different kinds of discrimination; and delineates diverse pathways by which discrimination can harm health, both outright and by distorting production of epidemiologic knowledge about determinants of population health. Three methods of studying health consequences of discrimination are examined (indirect; direct, at the individual level, in relation to personal experiences of discrimination; at the population level, such as via segregation), and recommendations are provided for developing research instruments to measure acute and cumulative exposure to different aspects of discrimination. PMID- 10379456 TI - New labour and the third way in the British National Health Service. AB - The British Labour Party claims that its policies are based on a "third way," new and distinct from both the old left and the new right. This article critically examines this claim with respect to health policy. After examining the Conservative legacy in the National Health Service and the evolution of Labour's health policy, the author introduces the concept of the "third way" and discusses the extent to which Labour's health policy can be seen in these terms, using the themes of spending, competition, accountability, and public health. There are many differences between the health policies of New and Old Labour, and some differences between those of New Labour and the Conservatives. Indeed, to a large extent Labour's health policy is built on the legacy of the Conservatives and is characterized by evolution. It is difficult to find any "big idea" or coherent philosophy behind the third way. Rather than being a new and distinctive approach rejecting both the old left and the new right, it seems to be a pragmatic pick and mix, attempting to combine the best from the market approach of the Conservatives and the hierarchical approach of Old Labour. PMID- 10379457 TI - The Mexican Social Security counterreform: pensions for profit. AB - The social security counterreform, initiated in 1997, forms part of the neoliberal reorganization of Mexican society. The reform implies a profound change in the guiding principles of social security, as the public model based on integrality, solidarity, and redistribution is replaced by a model based on private administration of funds and services, individualization of entitlement, and reduction of rights. Its economic purpose is to move social services and benefits into the direct sphere of private capital accumulation. Although these changes will involve the whole social security system--old-age and disability pensions, health care, child care, and workers' compensation--they are most immediately evident in the pension scheme. The pay-as-you-go scheme is being replaced by privately managed individual retirement accounts which especially favor the big financial groups. These groups are gaining control over huge amounts of capital, are authorized to charge a high commission, and run no financial risks. The privatization of the system requires decisive state intervention with a legal change and a sizable state subsidy (1 to 1.5 percent of GNP) over five decades. The supposed positive impact on economic growth and employment is uncertain. A review of the new law and of the estimates of future annuities reveals shrinking pension coverage and inadequate incomes from pensions. PMID- 10379459 TI - The health and safety concerns of immigrant women workers in the Toronto sportswear industry. AB - Immigrant women's conditions of work have worsened with new government and managerial strategies to restructure the Canadian apparel industry. Changes in occupational health and safety legislation have both given and taken away tools that immigrant women workers could use to improve the quality of their working lives. The author outlines a methodology for eliciting the health and safety concerns of immigrant women workers. PMID- 10379458 TI - A rise in the incidence of childhood cancer in the United States. AB - From the early 1980s to the early 1990s, the incidence of cancer in American children under 10 years of age rose 37 percent, or 3 percent annually. There is an inverse correlation between increases in cancer rates and age at diagnosis; the largest rise (54 percent) occurred in children diagnosed before their first birthday. Rates rose for all 11 states and cities included in the analysis. A jump in cancer rates for children born in 1982-83 was followed by a drop; but another abrupt rise for the 1986-87 birth cohort has been sustained thereafter. Results indicate that the rising childhood cancer rate represents a far more serious problem in the United States than previous reports have suggested. The methodology used here adds three additional states and cities, analyzes children under 10 rather than under 15, begins the analysis in 1980 rather than in 1973, and extends the study to 1993, which may partially account for the new findings. There are no apparent explanations for these trends, suggesting that researchers should analyze the data more fully and propose hypotheses on potential causes. One possible factor, fetal and infant exposure to low-dose radioactivity, is explored here. PMID- 10379460 TI - The rise, heyday, and incipient decline of specialization: hospitals in Denmark, 1930-1990. AB - Between 1930 and 1990 Denmark's hospital sector and hospital policy underwent radical changes. In 1930 the sector was dominated by many small hospitals, with care as the central task. By 1990 the number of hospitals had almost halved, specialization had developed, and diagnostic and therapeutic procedures were hospitals' most important functions. There have been many claims that the shape of the health care sector is determined by the development of medicine. This article demonstrates that changes in other areas of society have greatly influenced the development of the Danish hospital sector. In the 1930s and 1940s, the focus was on equity and specialization; in the 1950s, on growth, rationalization, and division of labor; in the 1960s, on growth and planning; and during the last decades, on management, productivity, and cost containment. Since 1980 the specialization, growth, and political acceptability of the specialized hospital sector have decreased, a change that can be characterized as the incipient decline of the specialized hospital sector. PMID- 10379461 TI - The politics of "natural" disaster: who made Mitch so bad? AB - The devastation in Central America following the 1998 hurricane (Hurricane Mitch) resulted more from economic and political policies than from "natural" disaster. Over the last 30 or 40 years, huge numbers of poor people in these countries have been forced off good, stable agricultural land onto degraded hillsides and into shanty towns constructed on floodplains--areas known to pose serious hazards of flooding and mudslides. This, together with the failure of impoverished countries to anticipate disaster through mass evacuations or to respond effectively to the hurricane's widespread damage--ensured the loss of thousands of lives. PMID- 10379462 TI - Epidemiology of non-steroidal anti-inflammatory drug damage to stomach and duodenum. AB - Conventional non steroidal anti-inflammatory drugs (NSAIDs) are probably responsible for a fifth to a quarter of cases of peptic ulcer bleeding, and for the same proportion of deaths. Predisposing factors include age, due probably to underlying susceptibility to ulcer, drug dosage, and the nature of NSAID. It is also likely that prior ulcer predisposes, but whether Helicobacter pylori infection without ulcer does so is unclear. Factors underlying the proportion of ulcer complications not associated causally with NSAID or aspirin treatment, two third of cases, are largely unknown. PMID- 10379464 TI - Defining patients at risk of non-steroidal anti-inflammatory drug gastropathy. AB - Non-steroidal anti-inflammatory drugs have long been known to cause gastro duodenal damage. However, all parts of the gastrointestinal tract may be affected, including the small intestine, colon and oesophagus. Non-steroidal anti inflammatory drugs can cause dyspeptic symptoms, erosions, ulceration, which may lead to haemorrhage or perforation, and a requirement for surgery. The purpose of this report is to assess risk factors which may lead to gastrointestinal damage and, thus, to identify those patients at greatest risk of non-steroidal anti inflammatory drug damage. Possible risk factors include age, sex, previous ulcer history, the presence of Helicobacter pylori, the type and severity of arthritis, individual non-steroidal anti-inflammatory drugs (dose, duration of treatment, route of administration), other debilitating diseases, smoking, alcohol, and the use of concomitant drugs. Risk of non-steroidal anti-inflammatory drug damage is higher in older patients (RR > 60 5.52; < 60 1.65), but there is no convincing sex difference. There is increased risk in patients with a previous history of peptic ulceration (RR first gastrointestinal event 2.39; subsequent gastrointestinal event 4.76), and in the first three months of treatment. Debate continues about the relevance of Helicobacter pylori, and this will be discussed in a later report. There is no strong evidence that patients with rheumatoid arthritis are more likely to have more trouble than those with osteoarthritis, but the former are more likely to require higher doses of non-steroidal anti inflammatory drugs. Highest risk non-steroidal anti-inflammatory drugs include azapropazone, ketoprofen and piroxicam, and those with least risk include ibuprofen, diclofenac and etodolac. There is an increased risk of gastrointestinal complications with relatively small-dose prophylactic aspirin. Other factors increasing the risk are smoking and the presence of chronic underlying respiratory and cardiovascular disease. Risk of gastrointestinal problems is increased with concomitant drugs, especially corticosteroids (RR 14.6 if given with non-steroidal anti-inflammatory drugs), but also with anticoagulants and some other drugs. The clinical importance of identifying possible risk factors lies in being aware of likely problem patients and in the use of safer non-steroidal anti-inflammatory drugs or combination therapy with protective drugs in these patients. PMID- 10379463 TI - Pathogenesis of non-steroidal anti-inflammatory drug gastropathy. AB - Although non-steroidal anti-inflammatory drugs (NSAIDs) may impair the defensive ability of the gastric mucosal barrier through topical actions, recent evidence suggests that microcirculation disturbance plays a pivotal role in the genesis of gastric mucosal damage. In particular, attention has been drawn to the role exerted by those cytokine (TNF alpha and IL-1 beta) and adhesion molecules (LFA 1, Mac-1, ICAM-1) that regulate interactions between leukocyte and endothelial cells leading to gastric microvessels occlusion and ischaemic/hypoxic endothelial epithelial cell damage. In recent years the role of prostaglandin synthesis inhibition in the pathogenesis of NSAID-gastropathy has been reconsidered, highlighting the immunomodulatory and pro-inflammatory consequences of prostanoids suppression. The awareness that mucosal damage is due to the non discriminatory effect of NSAIDs on cyclo-oxygenase (COX) isoenzymes, has lead to the development of more selective, safer, COX-2 inhibitors. On the other hand, the observation that NO exerts a predominant physiological role in the maintenance of mucosal integrity has raised the opportunity to develop a new generation of NO-releasing NSAID derivatives with a reduced gastrointestinal toxicity. PMID- 10379465 TI - Clinical symptoms, endoscopic findings and histologic features of gastroduodenal non-steroidal anti-inflammatory drugs lesions. AB - Gastrointestinal side effects of non-steroidal anti-inflammatory drugs are among the most prevalent iatrogenic gastrointestinal problems, due to the prevalence of problems, but even more because of the large and increasing population that is exposed. Dyspeptic complaints are prevalent, and cause discontinuation of the drug in 10% of patients. These symptoms fail, however, to correlate with endoscopic findings, and cannot be used to monitor patients for serious adverse events. The interventional measures include dose reduction, administration with food, concomitant use of antiacids or antisecretory drugs. Sometimes, changing the drug can help. Non-steroidal anti-inflammatory gastropathy comprises petechiae, haemorrhagic erosions and mucosal erythema. Such lesions are prevalent even in short term studies, but the clinical significance is uncertain, since an extrapolation to serious adverse events has not been documented. Histologically, a "chemical gastritis" has been described, but others fail to distinguish convincingly between Helicobacter pylori- and non-steroidal anti-inflammatory drug-induced inflammatory changes. Frank ulcerations are present in 20% of all long term users, with an even distribution between stomach and duodenum. Multiple ulceration in the antrum and prepyloric area are typical, but non-steroidal anti inflammatory drug ulcer may be indistinguishable from Helicobacter pylori ulcers and gastric cancers. Thus, even in the context of non-steroidal anti-inflammatory drug exposure, the possibility of malignancy must be ruled out by serial biopsies until complete healing. PMID- 10379466 TI - Non-steroidal anti-inflammatory drug gastropathy and Helicobacter pylori infection. AB - The interest in the interaction between NSAIDs and Helicobacter pylori derives its importance from its potential to provide a different strategy to combat the common problem of NSAID-related peptic ulcers and their life threatening complications. Studies assessing this subject have differed in almost every aspect of their methodology, including the definition of a NSAID user as well as the types, doses, duration, and the indications for NSAID use. They also differed in their end points, the tests for the assessment of Helicobacter pylori, and the regimes used for its eradication. However, NSAIDs and Helicobacter pylori are known to share a number of important pathogenic mechanisms, and the prevalence of the infection is high in cohorts of ulcer patients taking NSAIDs. Eradication of Helicobacter pylori using bismuth-based regimes has also been more successful and beneficial in preventing the occurrence or relapse of NSAID-related ulcers than regimes using proton pump inhibitors. PMID- 10379467 TI - Forthcoming non-steroidal anti-inflammatory drugs: are they really devoid of side effects? AB - Non-steroidal anti-inflammatory drugs are the most prescribed of the anti rheumatic drugs. The frequency and severity of their side effects on the gastrointestinal tract is a major health issue. A part of the toxicity of conventional non-steroidal anti-inflammatory drugs is due to their "topical" effect as well as their inhibition of cyclo-oxygenase-1. It has been suggested that the emergence of highly specific and selective cyclo-oxygenase-2 inhibitors will lead to significant decrease in gastrointestinal damage while maintaining or even improving therapeutic efficacy. Here I review the strength and weaknesses of conventional methods for assessing gastric and small intestinal safety of non steroidal anti-inflammatory drugs. The available safety data for a range of cyclo oxygenase-2 selective agents (meloxicam, nimesulide, celecoxib and vioxx) is reviewed. Short term endoscopy studies show minimal damage with these drugs and there is some data to suggest, at least for celecoxib and vioxx, that long term ingestion is not associated with significant gastric damage. Serious outcome studies are not available, but there is a suspicion that meloxicam may not be devoid of toxicity. Short term studies assessing intestinal permeability, which appear to give predictive information on the longer term small intestinal tolerability, show that meloxicam increases intestinal permeability while neither nimesulide or vioxx do so. Furthermore nimesulide does not cause non-steroidal anti-inflammatory drug-enteropathy when taken short term. So far the more selective cyclo-oxygenase-2 inhibitors are living up to their promise. PMID- 10379468 TI - Non-steroidal anti-inflammatory drugs and gastrointestinal bleeding. AB - Non-steroidal anti-inflammatory drug use carries the risk of gastrointestinal complications (1% over 6 months) which is increased by a factor of 4 to 5, although strong differences are observed between different non-steroidal anti inflammatory drugs. This risk is present in both the upper and lower gastrointestinal tract which indicates that non-steroidal anti-inflammatory drugs induces bleeding from both peptic ulcer and non-peptic ulcer sources. Symptoms are poor predictors of serious lesions and complications, which may occur without previous symptoms. At present, risk factors for non-steroidal anti-inflammatory drug-associated upper gastrointestinal bleeding are well defined and include ulcer or complication history, age, high non-steroidal anti-inflammatory drug dose, combination with corticosteroid and warfarin. Helicobacter pylori infection is not considered a risk factor for complications in non-steroidal anti inflammatory drug users. There is a high prevalence of over-the-counter non steroidal anti-inflammatory drug (especially aspirin) use among those presenting with gastrointestinal complications. Prophylactic aspirin regimens increase the risk of gastrointestinal bleeding. The potential beneficial effect of nitrate treatments (nitric oxide donors) in low dose aspirin users deserves further study. The mechanisms involved in the induction of gastrointestinal bleeding by non-steroidal anti-inflammatory drugs are poorly understood. Platelet activity inhibition associated with an abnormal, but reversible, prolongation of the bleeding time in susceptible individuals using aspirin might be a mechanism affecting no more than a third of patients with gastrointestinal bleeding. PMID- 10379469 TI - Pharmacological approach to the prevention of non-steroidal anti-inflammatory drug-induced gastropathy. AB - Non-steroidal anti-inflammatory drugs can provoke gastric damage by multiple interactive mechanisms. These processes include topical irritant actions that disrupt the epithelial barrier, which allows the back-diffusion of acid into the mucosa. The inhibition of the cyclo-oxygenase isoform, cyclo-oxygenase-1 also promotes gastric injury, such effects involving the microcirculation. These mechanisms of mucosal damage synergistically interact to cause more extensive injury, which can be attenuated by antisecretory agents or by mucosal protective agents such as the synthetic prostanoid, misoprostil. In addition, agents that release nitric oxide may prevent such mucosal damage. However, the development of the novel anti-inflammatory drugs, the cyclo-oxygenase-2 selective agents, that inhibit the formation of prostanoids at inflammatory sites, but not the endogenous protective prostanoids in the stomach formed by cyclo-oxygenase-1, has provided a highly effective therapeutic approach to minimising of gastric damage. PMID- 10379470 TI - Non-steroidal anti-inflammatory drug gastropathy: clinical results with antacids and sucralfate. AB - The efficacy of antacids in the short- and long-term treatment of peptic ulcers, has suggested a possible use in the prevention and in the treatment of non steroidal anti-inflammatory drug related gastroduodenal lesions. In short-term prevention studies, significant protection against ASA-related lesions was observed when antacids at high-dose were given before the administration of the offending drug. To the contrary, antacids at low dose did not prevent ASA-induced lesions of gastric and duodenal mucosa. As for long-term prophylaxis, no clinical effect was observed. In the treatment of non-steroidal anti-inflammatory drug related mucosal lesions in patients who were able to discontinue the offending drugs, antacids proved of some use, when compared with placebo, but were significantly less effective than H2 blockers, as cimetidine. Sucralfate is an effective antiulcer drug thought to provide cytoprotective action. Although initial studies utilizing sucralfate for protection against short-term aspirin administration were encouraging, longer term studies (more than 7 days) were generally disappointing. A comparative study with misoprostol demonstrated that the PGE1 analogue was far superior for the prevention of non-steroidal anti inflammatory drugs ulcers, and that ulceration rates in the sucralfate group were equivalent to rates in the placebo group. As far as the treatment of non steroidal anti-inflammatory drug-related mucosal lesions is concerned, sucralfate proved superior to placebo, similar to ranitidine, but significantly less effective than omeprazole. PMID- 10379471 TI - Non-steroidal anti-inflammatory drug gastropathy: clinical results with misoprostol. AB - The aim of the review was to examine the effectiveness of misoprostol as co therapy in the prevention of non-steroidal anti-inflammatory drug-induced gastrointestinal mucosal injury. The outcomes assessed were the number of patients in whom a gastric ulcer developed, the number of patients in whom gastric lesions developed. The number of patients in whom a duodenal ulcer developed and the number of patients in whom duodenal lesions developed. The weighted average baseline risks for gastric ulcers were found to be 3.8% and 6.8% with short- and long-term non-steroidal anti-inflammatory drug treatment, respectively. Misoprostol treatment resulted in a significant (p < 0.05) risk reduction of gastric ulcer in the short-term (pooled RD = 13.3%; 95% confidence interval -25.7%, -0.9%) and in the long-term (RD = -8.4%; 95% confidence interval -17.7%, -1.0%) non-steroidal anti-inflammatory drug treatment. The weighted average baseline risks for duodenal ulcers were found to be 3% and 4% with short- and long-term non-steroidal anti-inflammatory drug treatment, respectively. Misoprostol did not significantly reduce the risk of duodenal ulcers with short treatment (RD = -2.0%; 95% confidence interval -5.7%, 1.6%). Treatment was effective in the long-term non-steroidal anti-inflammatory drugs therapy (RD = 3.4%; 95% confidence interval -5.8%, -0.1%, p < 0.001). The number of patients needed to be treated to prevent 1 person from developing a gastric ulcer within 2 weeks of non-steroidal anti-inflammatory drug therapy was found to range from 35, when baseline risk was 3%, to 3 when the baseline risk was higher, say 40%. The corresponding number of patients needed to be treated for gastric ulcer prevention in long-term studies ranged from 47 to 5. The number of patients needed to be treated for prevention of duodenal ulcer with misoprostol in short term studies ranged from 36 to 4 and in the long-term trials form 47 to 8. In conclusion, a rational approach to treat only high risk patients is recommended. PMID- 10379472 TI - Prevention and treatment of non-steroidal anti-inflammatory drug-induced gastro duodenal damage: rationale for the use of antisecretory compounds. AB - Gastro-duodenal mucosa possesses an array of defensive mechanisms and non steroidal anti-inflammatory drugs have a deleterious effect on most of them. This results in a mucosa less able to cope with even a reduced acid load. The presence of acid appears to be a conditio sine qua non for non-steroidal anti-inflammatory drug-injury, which is in fact pH-dependent. The acute damage induced by acid non steroidal anti-inflammatory drugs, like aspirin, can be markedly reduced or even prevented by raising intragastric pH with antacids or antisecretory compounds. Animal studies have clearly shown that not only the degree, but also the duration, of acid inhibition is an important factor for prevention of non steroidal anti-inflammatory drug-induced mucosal damage. As a consequence, proton pump inhibitors (PPIs) appear to be more effective that H2-receptor antagonists both in preventing and treating gastro-duodenal lesions. While acid suppression seems to be the only effective mechanism for ulcer healing, prevention of non steroidal anti-inflammatory drug-injury might also rely on the mucosal protective activity of these compounds. Clinical pharmacological studies, performed in healthy volunteers, have shown that--as in laboratory animals--elevation of intragastric pH by means of antacids or antisecretory compounds protects against acute NSAID-induced damage. Unlike H2-blockers, PPIs protect from non-steroidal anti-inflammatory drug-injury not only the duodenum, but also the stomach, where the majority of mucosal lesions are usually located. Although elevation of intragastric pH affects non-steroidal anti-inflammatory drug pharmacokinetics and pharmacodynamics in laboratory animals, a lack of drug-to-drug interaction between PPIs and some of these compounds has been reported in humans. To summarize, clinical and experimental pharmacology support the use of PPIs for the prevention and treatment of non-steroidal anti-inflammatory drug-induced gastro duodenal damage. Acid suppression could, however, represent only one of the many mechanisms by which these compounds protect gastro-duodenal mucosa. Further studies are, therefore, needed to better elucidate the respective role of the various pharmacological actions in their mucosal protective activity as well as to assess the clinical relevance of each of them. PMID- 10379473 TI - Non-steroidal anti-inflammatory drug gastropathy: clinical results with H2 antagonists and proton pump inhibitors. AB - While the most effective strategy to prevent non-steroidal anti-inflammatory drug related gastrointestinal toxicity is not to prescribe the medication, this option is often impractical. The use of specific agents to heal mucosal lesions or to prevent non-steroidal anti-inflammatory drug toxicity, has focused upon two approaches: replacement of prostaglandin deficiency and inhibition of acid secretion. Acid suppression with traditional ulcer healing doses of H2-blockers is effective in the cure of gastric and duodenal ulcers upon discontinuation of the offending drug. In the event the non-steroidal anti-inflammatory drug must be continued, the use of H2-RAs is associated with a slight decrease in the healing rate. In long-term prevention studies, H2-blockers significantly reduce duodenal ulcer rates, but are ineffective in reducing gastric ulceration. More potent acid inhibition with double-doses of H2-blockers may also reduce the risk of gastric (famotidine 80 mg) and duodenal ulcers (famotidine 80 mg or ranitidine 600 mg daily). Proton pump inhibitors (omeprazole 20-40 mg, lansoprazole 30 mg daily) appear more effective in healing gastric and duodenal ulcers in patients continuing the offending drug. Comparative studies of omeprazole vs ranitidine, misoprostol and sucralfate show a therapeutic gain in favour of the proton pump inhibition, ranging from 10 to 40%. In long-term prevention studies, omeprazole (20 mg daily) and pantoprazole (40 mg daily) have also been shown to reduce the risk of gastric and duodenal ulcers. Comparative studies of omeprazole (20 mg daily) vs ranitidine (150 mg daily) and misoprostol (200 micrograms daily) showed that after 6 months' follow-up the proton pump inhibition was significantly superior to control drugs in reducing the risk of both gastric and duodenal ulcer. PMID- 10379474 TI - Pharmacoeconomic aspects of non-steroidal anti-inflammatory drug gastropathy. AB - Non-steroidal anti-inflammatory drugs are commonly used to reduce inflammation and pain associated with arthritis. However, non-steroidal anti-inflammatory drugs induce gastrointestinal side-effects such as dyspeptic symptoms, duodenal or gastric ulcers and, in some cases, serious complications. The aim has been to compare the benefits with the drawbacks of non-steroidal anti-inflammatory drug treatment using a hypothetical population representing patients with arthritis. A problem description was made on the basis of a literature review, and a simple and hypothetical health economic model was constructed. Including direct and indirect costs, the annual total costs in Sweden for gastrointestinal side effects per non-steroidal anti-inflammatory drug user were estimated to be 3,420 SEK (438 US$), and the approximated costs of arthritis were 60,000 SEK (7,692 US$). The benefits of non-steroidal anti-inflammatory drug treatment were found to outweigh the drawbacks if the patient's arthritis symptoms, expressed as a difference in utility value between having and not having symptoms of arthritis, are improved by 6% or more. Costs for non-steroidal anti-inflammatory drug induced gastrointestinal side-effects should be evaluated in relation to the benefits of non-steroidal anti-inflammatory drugs in the treatment of inflammation and pain. A simple modelling approach indicated that treatment with non-steroidal anti-inflammatory drugs may be highly cost-effective as both the clinical and economic benefits for patients responding to such treatment out weighed possible drawbacks. PMID- 10379476 TI - Cystic neoplasms of the pancreas. PMID- 10379475 TI - A clinical approach to management of patients with non-steroidal anti inflammatory gastropathy. AB - The treatment of a peptic ulcer occurring in a patient who is taking non steroidal anti-inflammatory drugs depends on whether or not the patient can readily stop taking the non-steroidal anti-inflammatory drug. If they can, healing is generally rapid, and can be achieved with any effective ulcer-healing agent. When the non-steroidal anti-inflammatory drug cannot be easily stopped, ulcer healing is slower and the treatment of choice is to heal the ulcer with a proton pump inhibitor. The risk of ulceration in patients taking non-steroidal anti-inflammatory drugs can be reduced by two main strategies: the choice of non steroidal anti-inflammatory drug and its dosage on the one hand, and the use of prophylactic co-therapy on the other. The two are not of course mutually exclusive. It is now clear that not all non-steroidal anti-inflammatory drugs are equally damaging. Several studies have shown that the shorter half life drugs (at least in their recommended dosages) are generally less ulcerogenic. There is clear dose-dependence, so the drugs should be used at the lowest effective dose, and non-steroidal anti-inflammatory drugs should not be given in combination without careful weighing of risks and benefits. Giving either omeprazole or misoprostol concurrently with a non-steroidal anti-inflammatory drug substantially reduces the risk of ulceration. Full dosage histamine H2-receptor antagonists give good protection against non-steroidal anti-inflammatory drug associated duodenal ulcers, but two large trials with ranitidine showed no protection against gastric ulcer. One study of double dosage famotidine did show a reduction in gastric ulcer incidence as well. Recently, two large randomized trials have compared omeprazole 20 mg daily head-to-head with ranitidine and misoprostol. Overall, the proton pump inhibitor was more effective than the other two for ulcer prevention, although it is interesting that erosions seemed to be prevented better by the prostaglandin. The biggest challenge for clinical judgement is when to use prophylactic co-therapy. Patients for whom this should be especially considered are those who have had a prior ulcer, the elderly, those needing higher non-steroidal anti-inflammatory drug dosage or co-therapy with vascular-protective aspirin, and those whose other medical conditions make them less likely to survive a gastrointestinal haemorrhage or perforation. PMID- 10379477 TI - Food allergy and Helicobacter pylori infection. AB - BACKGROUND: Most antigens reach the immune system through mucosae. Gastrointestinal mucosa is a barrier for alimentary antigens. Inflammatory processes, such as Helicobacter pylori-associated gastritis, could alter the integrity of the gastric barrier, increase the mucosal permeability, and enhance crossing of food antigens which may stimulate allergic reactions. PURPOSE: The aim of this study was to establish whether patients with symptomatic food allergy and detectable immunoglobulin E (IgE) to alimentary antigens were infected by Helicobacter pylori more often than controls, and to determine the phenotype of the infecting Helicobacter pylori. PATIENTS AND METHODS: Thirty-eight consecutive patients with symptomatic food allergy and serum IgE to alimentary antigens, and 53 consecutive age-matched controls (subjects without food allergy and detectable levels of IgE anti-alimentary antigens) living in the same area and attending the same institution were investigated serologically to determine the prevalence of Helicobacter pylori infection, and an immune response to CagA, a marker of the most pathogenic strains. IgE to alimentary allergens were measured by a commercial kit. RESULTS: The prevalence of Helicobacter pylori infection in patients with food allergy and controls was similar (42.1% and 47.1%, respectively). Anti-CagA antibodies in Helicobacter pylori-infected persons were detected in 62.5% of patients with food allergy, and 28.0% of controls (p = 0.030, odds ratio = 4.29, RR = 2.23). The mean IgE level to the most common alimentary antigens was increased in CagA-positive, with respect to the CagA negative, patients. CONCLUSIONS: The enhanced mucosal and inflammatory lesions commonly found in individuals infected by CagA-positive Helicobacter pylori strains could increase the epithelial permeability and render non-selective the passage of allergens which, in atopic persons, could directly stimulate an IgE response. Infection by CagA-positive Helicobacter pylori may increase the risk of food allergy. PMID- 10379478 TI - Dysplastic changes in gastric fundic gland polyps of patients with familial adenomatous polyposis. AB - BACKGROUND: Fundic gland polyps are the most common gastric lesion in patients with familial adenomatous polyposis and are traditionally considered a condition with no malignancy potential. However, some reports have recently questioned this view. AIMS: To prospectively evaluate their prevalence and the associated dysplastic/malignant changes in a series of affected patients. PATIENTS AND METHODS: Thirty-seven affected patients were carefully investigated by upper endoscopy over a three-year period. Multiple (at least 10) complete excisions of any representative polyp of the body-fundus were performed and a thorough pathological search for microscopic adenomatous/dysplastic changes carried out. RESULTS: Of 37 patients, 19 (51.3%) showed gastric fundic gland polyposis and 18 of them gave consent for polypectomies. Overall, 425 endoscopic polypectomies were performed, with a mean of 23.6 +/- 14.6 per patient. At pathology, all excised polyps of the body-fundus were found to be fundic glandular. Microscopic adenomatous changes within such polyps were identified in 8 (44.4%) patients. All the adenomatous foci revealed mild dysplasia with no case of severe atypia or carcinoma. Patients with microadenomas showed a significantly higher total number of gastric polyps compared with those without microadenomas (p < 0.03). No other differences between the two groups were observed. Two further patients presented microadenomas in apparently normal antral mucosa and one also showed a 6 mm antral adenoma with mild dysplasia. Finally, the search for Helicobacter pylori was always negative. CONCLUSIONS: Patients with familial adenomatous polyposis and gastric fundic gland polyps have a high prevalence of microscopic adenomatous foci within such lesions; nevertheless, these foci seem not to be associated with signs of severe atypia or carcinoma. Moreover, microadenomas are ubiquitous throughout the stomach, as well as in the rest of the gut, and their natural history is still undefined. Thus, their malignancy potential remains uncertain. More extensive follow-up is warranted to better investigate the long-term biological behaviour of these lesions but, at present, our data do not support the need for a change in the usual intervals of upper endoscopy surveillance in familial polyposis patients with or without gastric fundic glands polyps. PMID- 10379479 TI - Family linkage study of the T-cell receptor genes in coeliac disease. AB - BACKGROUND: The susceptibility to coeliac disease is genetically determined by possession of certain HLA DQ alleles, together with a one or more non-HLA genes. The central role of the T-cell receptor in disease pathogenesis makes the T-cell receptor genes strong candidates as disease susceptibility genes, and previous studies had provided equivocal ambiguous results. METHODS: A pedigree based genetic linkage study was used to determine if any of the T-cell receptor genes have a role in the genetic aetiology of coeliac disease. Intragenic microsatellite markers were used to study T-cell receptor alpha, beta, and delta, while gamma was studied using two flanking microsatellites D7S484 and D7S629. RESULTS: Conventional linkage analysis was performed using the MLINK computer package. Model-free linkage analysis was performed using MFLINK. No evidence of linkage between coeliac disease and the T-cell receptor genes was found in these pedigrees. CONCLUSIONS: Mutations in the T-cell receptor genes are not implicated in the genetic aetiology of coeliac disease. PMID- 10379480 TI - Contribution of molecular genetics to gastroenterology: the case of coeliac disease. PMID- 10379481 TI - Effect of moderate exercise on Crohn's disease patients in remission. AB - BACKGROUND: Physical exercise may exacerbate the disturbed homeostasis of Crohn's disease patients. AIM: To examine the effect of moderate physical exercise on gastrointestinal function in a group of Crohn's disease patients in remission. PATIENTS AND METHODS: The effect of one-hour's exercise at a maximum of 60% oxygen consumption was evaluated in six males with ileal Crohn's disease in remission on orocaecal transit time (breath test to lactulose), intestinal permeability (6-hours' urinary excretion of a sugar mixture of lactulose/mannitol), polymorphonuclear leucocytes function (peripheral blood chemiluminescence), lipoperoxidation (plasma malondialdehyde) and antioxidant trace elements (urinary and plasma zinc and copper concentrations). Six healthy age-matched subjects served as controls. RESULTS: Exercise did not elicit subjective symptoms or changes in intestinal permeability and lipoperoxidation. Orocaecal transit time increased after exercise in Crohn's disease patients (72 min +/- 30 vs 100 min +/- 34) with no significant difference from controls (77 min +/- 20 vs 83 min +/- 23). Neutrophils, primed pre-exercise in Crohn's disease patients showed an increased post-exercise chemiluminescence similar to controls. Zinc urinary output significantly increased after exercise in Crohn's disease patients and remained unchanged in control subjects. CONCLUSIONS: Moderate aerobic exercise has no significant effect on the gastrointestinal parameters examined. However, basal neutrophil activation and exercise in Crohn's disease patients may trigger an excessive production of oxygen metabolites. Moreover, exercise may contribute to an increased risk of zinc deficiency. PMID- 10379482 TI - Retreatment of non-responder or relapser chronic hepatitis C patients with interferon plus ribavirin vs interferon alone. AB - BACKGROUND AND AIM: Interferon-alpha treatment of chronic hepatitis C is beneficial in only 20-30% of patients. This study evaluates if combination therapy with Interferon-alfa plus ribavirin is effective in inducing a response in patients who did not respond to, or relapsed after, a standard Interferon-alfa treatment. PATIENTS AND METHODS: A total of 88 patients, 49 non-responders and 39 relapsers to previous Interferon-alfa therapy, were randomized to receive either natural Interferon-alfa (6 MU t.i.w.) plus ribavirin (1000 mg/daily) or natural Interferon-alfa alone (6 MU t.i.w.) for 6 months. All were followed for 12 months after stopping therapy. Serum aminotransferase levels were assessed monthly and HCV RNA was evaluated by RT-PCR (Amplicor, Roche) at end of therapy and the end of follow-up. RESULTS: After treatment, a higher response rate defined as return to normal of aminotransferases and absence of serum HCV RNA was observed among patients treated with Interferon-alfa-ribavirin: 4/28 (14%) vs 1/21 (5%) non responder patients and 9/19 (47%) vs 5/20 (25%) in the relapsers group. At the end of follow-up, a sustained response was found only in the combination treatment group: 4% and 32% in non-responder and relapser patients, respectively. CONCLUSIONS: Our results suggest that retreatment with natural Interferon-alfa plus ribavirin is more effective than Interferon-alfa alone in increasing the response rate in patients with chronic hepatitis C who relapse after a previous standard IFN treatment whereas it is less effective in non-responder patients. PMID- 10379483 TI - Interferon + ribavirin combination therapy. Open new approach to treatment of patients with hepatitis C virus. PMID- 10379484 TI - Epidemiologic and genetic factor in colorectal cancer: development of cancer in dizygotic twins in a family with Lynch syndrome. AB - Human tumours usually develop due to a close interaction between environmental and genetic factors. This concept applies also to well defined genetic diseases such as Hereditary Nonpolyposis Colorectal Cancer (HNPCC or Lynch syndrome), which is featured by early onset tumours of the large bowel (and other target organs), striking aggregation of neoplasms in families, and vertical transmission consistent with an autosomal dominant pattern of inheritance. As a further example of gene/environment interaction, we report on a Hereditary Nonpolyposis Colorectal Cancer family in which two dizygotic twins were affected by cancer of the large bowel. One of the twins was slightly overweight and showed many common risk factors for colorectal carcinoma; he developed a Dukes' C lesion at the age of 52 years. The other twin was not overweight and was much less exposed to exogenous risk factors; a Dukes' B carcinoma was diagnosed at age 60, during a control endoscopy. This anedoctal report suggests that diet and lifestyle are of relevance also in patients with genetically determined tumours of the large bowel. It follows that the control of these environmental factors might be associated with a delay of tumour occurrence and possibly with a less aggressive tumour behaviour. PMID- 10379485 TI - Environment and genes in the development of colorectal cancer. PMID- 10379486 TI - AISF practice guidelines for portal hypertension. Committee for Portal Hypertension of the Italian Association for the Study of the Liver (AISF). PMID- 10379487 TI - Tumour necrosis factor alpha--mediator of apoptosis and cell proliferation of hepatocytes. AB - In recent years, the intracellular pathways activated by the tumour necrosis factor have been described in great detail. Adaptor molecules which bind to the intracellular domain of the tumour necrosis factor receptor 1 are able to either induce apoptosis or to activate signals which trigger cell proliferation, anti apoptotic mechanisms or the inflammatory response of hepatocytes. In different animal model the tumour necrosis factor-dependent mechanisms have been defined. Therefore, the present review deals with the molecular mechanisms of tumour necrosis factor-dependent signalling. Additionally, the role of tumour necrosis factor during liver regeneration after partial hepatectomy and models of tumour necrosis factor-dependent liver cell damage will be discussed. PMID- 10379488 TI - Crohn's disease: the case for bacteria. AB - At the present time, there is no convincing indication that Crohn's disease is a bacterial disease, although an association with mycobacteria has been hypothesised for many years. The hypothesis that bacteria could be the cause, or at least an important concause of Crohn's disease is supported by several experimental and clinical observations: animals kept in a germ-free environment fail to develop intestinal inflammation; bacteria are the cause of human and animal intestinal diseases similar to Crohn's disease; luminal content is necessary for causing gut lesions; and, moreover, antibiotics are successfully used in the treatment of Crohn's disease. Bradford Hill criteria recently used to assess a causal relationship for Helicobacter pylori and peptic ulcer can be applied for establishing or excluding a causality between mycobacteria and Crohn's disease. Of these criteria, only biological plausibility, coherence and analogy are satisfied. However, failure to identify a specific pathogen does not exclude a possible role for bacteria in causing Crohn's disease lesions and symptoms. Pathogenic or commensal enteric bacteria could overinfect the primary lesions, leading to chronic intestinal inflammation in genetically susceptible hosts. Another possibility is that components of the normal intestinal flora could acquire pathogenic characteristics. PMID- 10379489 TI - Crohn's disease: the case for measles virus. AB - Crohn's disease has the epidemiological and pathological hallmarks of an infection with a long natural history. Its emergence in developed countries in the middle of the 20th Century represents an instant in the continuum of human evolution, indicating either a new infection or, as with poliomyelitis, a changing pattern of exposure to a common childhood pathogen. Both short- and long term outcomes from viral infection are largely dependent upon age and dose of exposure. We and others have suggested that measles virus may be causally related to Crohn's disease, and that the associated risk is an atypical pattern of exposure. Early, intensive, and concurrent infections have been identified as risks for subacute sclerosing panencephalitis, a delayed sequelae to measles virus infection, possibly through a process of high zone immunological tolerance and persistent infection. The data for Crohn's disease suggest that persistent infection may follow early low dose exposure and low zone immunological tolerance. The changing pattern of measles virus exposure this century would be consistent with a shift towards lower dose of infection. Such an exposure would also be consistent with persistence of the virus at very low copy number within discrete foci of granulomatous inflammation. The ability of measles virus to profoundly disrupt mucosal immune responses may provide the human counterpart of the cytokine-gene knockout. PMID- 10379490 TI - Measles, mycobacterium paratuberculosis and Crohn's disease. PMID- 10379491 TI - No evidence of an impaired nutritional status in the early stage of schistosomiasis. PMID- 10379492 TI - Seven days of ranitidine bismuth citrate plus two antimicrobials for Helicobacter pylori eradication. PMID- 10379493 TI - Helicobacter pylori eradication using one-week low-dose lansoprazole plus amoxycillin and azithromycin: failure of a "promising" association. PMID- 10379494 TI - Anorectal reconstruction after abdominoperineal excision: a state-of-the-art alternative to a conventional colostomy. PMID- 10379495 TI - Venous Doppler velocimetry in relationship to central venous pressure and heart rate during hypoxia in the ovine fetus. AB - OBJECTIVE: Characteristic changes in ductus venosus (DV) blood velocity and pulsations in the umbilical vein (UV) have been described during imminent fetal asphyxia. The aim of this study was to examine fetal venous blood velocity in relationship to pressure gradient across the DV during hypoxia in a fetal lamb preparation. METHODS: In general anesthesia, a cesarean section was performed on seven pregnant ewes, the fetus was exteriorized and put into a heated waterbath with uninterrupted umbilical circulation. Pressure measurements in the UV and inferior vena cava (IVC) were performed with the catheter tips on both sides of the DV. Fetal hypoxemia was induced by giving the ewe 12% oxygen in inhaling air. Pressure across the DV and Doppler velocimetry were repeatedly measured during hypoxemia. Blood velocity was recorded in the DV and UV by Doppler ultrasound. RESULTS: Before hypoxia the median pressure gradient across the DV was in systole 1 mmHg and 0.31 mmHg in end-diastole and during hypoxemia 1.5 mmHg and zero, respectively. The pressure difference across the DV was constant during hypoxemia irrespective of the presence of umbilical venous pulsations or heart rate. IVC pressure was greatly influenced by fetal heart rate (FHR). A small but linear fall in systolic IVC pressure was seen with increasing FHR. In end-diastole the IVC pressure changed in a parabolic fashion, with increasing pressure during brady- and tachycardia. Pulsations in the UV also showed a parabolic relationship to FHR and central venous pressure. DV end-systolic and end-diastolic blood velocity changed during hypoxemia in direct relationship to FHR and central venous pressure, but without direct relationship to fetal blood gases. CONCLUSION: The pressure gradient across the DV is constant during hypoxemia. Changes in central and umbilical venous pressure are directly related to FHR. Umbilical venous and DV blood velocity changed in direct relationship to FHR and central venous pressure. PMID- 10379496 TI - Volume and vascularity of the yolk sac studied by three-dimensional ultrasound and color Doppler. AB - The aim of our study was to assess the volume of the gestational sac and yolk sac throughout the first trimester of pregnancy, and to establish the relationship between the yolk sac volume measurements and vascularity visualization rates. Eighty women with uncomplicated singleton pregnancies between 5 and 12 weeks were evaluated by three-dimensional and color Doppler ultrasound (Combison 530, Kretztechnik). Regression analysis revealed exponential rise of the gestational sac volume with gestational age throughout the first trimester. An exponential rise of the yolk sac volume was noticed between gestational weeks 5 and 8, followed by gradual increase of the yolk sac volume between the gestational weeks 8 and 10. After reaching the plateau from 10 to 11 weeks, yolk sac volume started to decrease. The highest visualization rates for the yolk sac vessels were obtained between gestational weeks 7 and 8. When yolk sac reached the maximum size between 10 and 11 weeks, reduced vascularity was demonstrated. Three dimensional ultrasound allowed estimation of the gestational sac and yolk sac volumes throughout the first trimester of pregnancy. Both of these measurements seem to be useful prognostic parameters for the pregnancy outcome. The combination of functional and volumetric data provides much useful information on early human development. PMID- 10379497 TI - Three-dimensional transvaginal ultrasound improves measurement of nuchal translucency. AB - The aim of the study was to correlate intra-observer reproducibility of the nuchal translucency measurements by two-dimensional and three-dimensional transvaginal ultrasound. Examinations were performed on 120 women undergoing ultrasound screening at 10 to 14 weeks' gestation. They were examined by two experienced ultrasonographers using both methods two times consecutively. Statistical analysis for the assessment of intra-observer reproducibility was paired t-test. Nuchal translucency measurements were obtained in 100% of cases with three-dimensional sonography compared to only 85% with two-dimensional sonography. Better intra-observer reproducibility was obtained for three dimensional than for two-dimensional ultrasound. Three-dimensional transvaginal ultrasound improves accuracy of nuchal translucency measurement allowing appropriate mid-sagittal section of the fetus and clear distinction of the nuchal region from the amniotic membrane. PMID- 10379498 TI - Is the liver of the fetus the 4th preferential organ for arterial blood supply besides brain, heart, and adrenal glands? AB - In this study we compared the distribution of blood flow to the liver in growth retarded fetuses whose estimated weight was < 5th centile with normal-weight fetuses. As expected, the relative venous blood flow to the liver was reduced, with blood flowing preferentially through the ductus venosus. However, the total blood supply seemed to be maintained by a concomitant, significant increase in arterial blood flow through the hepatic artery. Absolute flow velocities such as the peak, minimum diastolic and temporal average velocities were changed, as was the flow waveform. Effectively, the deficiency in venous supply was made up for by an increase in arterial blood flow. This compensatory effect may be crucial for maintaining liver function in times of low portal venous blood supply. It thus makes sense to regard the liver as the fourth preferential organ for arterial blood supply in the compromised fetus, besides heart, brain, and adrenals. PMID- 10379499 TI - Postnatal change of renal artery blood flow velocity and its relationship with urine volume in very low birth weight infants during the first month of life. AB - Although the renal artery blood flow velocity has been investigated recently using the ultrasound Doppler method, little is known about the longitudinal change of renal artery blood flow velocity and its relationship with urine volume in very low birth weight infant. Thus, we measured renal artery blood flow velocities by means of the pulse Doppler method in 28 very low birth weight infants. Maximum, minimum, and mean blood flow velocities were determined at postnatal days 0, 1, 2, 3, 4, 5, 6, 13, 20, and 27. The resistance index was also calculated. The maximum and mean blood flow velocities increased gradually after birth, and were significantly higher at 13, 20, and 27 days after birth. The minimum blood flow velocity and the resistance index were relatively constant during the study period. The mean blood flow velocities were also analyzed for any correlation with urine volume. There was a poor correlation between urine volume (ml/kg/day) and mean blood flow velocity (cm/s) (Y = 2.38X + 57.4, Y: urine volume, X: mean blood flow velocity, n = 161, r = 0.338, P < 0.01). However, if the mean renal artery blood flow velocity was less than 10 cm/s, oliguria was observed in most cases. The measurement of the renal artery blood flow velocities appears to be useful in understanding the background condition of renal function in very low birth weight infants. PMID- 10379500 TI - Late hyporegenerative anemia in neonates with rhesus hemolytic disease. AB - This study was conducted to determine the risk factors of the late hyporegenerative anemia in Rh-isoimmunized infants. Data on 36 infants with rhesus hemolytic disease were analyzed. The mean gestational age and birth weight were 36 +/- 1.3 weeks and 2837 +/- 403 grams respectively. Twenty-seven infants (75%) received between 2 and 8 intravascular intrauterine blood transfusions. Fourteen infants (39%) required simple packed red blood cell transfusions and 11 infants (31%) required exchange blood transfusion in the immediate postnatal period. Thirty infants (83%) developed late anemia and required blood transfusions at a mean postnatal age of 43.3 +/- 15.7 days. Sixty-four percent of infants who had exchange blood transfusions did not develop late anemia, while 92% of infants who did not require exchange blood transfusion developed late anemia, and the difference was statistically significant (P = 0.035). Serum erythropoietin levels were determined in 8 infants immediately before simple transfusion for late anemia. The media serum erythropoietin level was 21.2 mU/ml, ranging between less than 10 to 114.2 mU/ml. We conclude that late hyporegenerative anemia is common among Rh isoimmunized infants, regardless of the intravascular intrauterine transfusion. Exchange blood transfusion was associated with less occurrence of late anemia. PMID- 10379501 TI - Neonatal screening for congenital cytomegalovirus infections. AB - We evaluated a screening program for the detection of congenital cytomegalovirus in 3075 unselected pregnant women. From each live-born child urine for CMV culture was collected within 7 days after birth. Each fetus expelled after a spontaneous second trimester abortion and each stillborn infant were also evaluated for a possible congenital CMV infection. For each congenital infection stored maternal sera were analysed to determine whether maternal infection was primary or recurrent. Fifteen out of the 3075 pregnancies studied resulted in a congenitally infected infant (0.49%). Nine maternal CMV infections were primary infections; five were recurrent infections, and in one case the type of infection could not be determined. Three congenital infections resulted in severe sequelae, leading to the termination of pregnancy in two instances and to neonatal death in one case. One of these severe fetal infections was due to a recurrent maternal infection. Follow-up of the other 12 neonates demonstrated hearing disorders in two children. One was born after a primary maternal infection and one after a recurrent maternal infection. We conclude that congenital CMV infections occurs in 0.49% of all pregnancies in the population studied. Twenty percent of the congenitally infected infants present severe sequelae at birth or during pregnancy, and an additional 17% have audiological deficits at 1 year of age. Severe sequelae may occur after both primary and recurrent maternal CMV infection. PMID- 10379502 TI - Familial perinatal hemochromatosis: a disease that causes recurrent non-immune hydrops. AB - Perinatal hemochromatosis is a rare disorder with an enormous iron overload in the parenchymal organs, especially the liver, pancreas, heart and endocrine glands. Elements of the reticuloendothelial system are relatively spared. The clinical course is rapidly progressive and the disease is invariably fatal. Several siblings are described in the literature. Herein, we describe one pair of full siblings affected by the disease, wherein the clinical presentation was hydrops. We suggest that hemochromatosis should be considered in the differential diagnosis of hydrops fetalis. PMID- 10379503 TI - A case of large placental chorioangioma with non-immunological hydrops fetalis. AB - A 34-year-old Japanese woman (gravida 2, para 2) with polyhydramnios and non immunological hydrops fetalis was referred to our department at 32 weeks of gestation. On admission, the blood pressure was 120/60 mmHg and there was no pitting edema of the lower extremities. An ultrasound examination disclosed a large placental tumor 5.8 cm x 4.4 cm x 4.8 cm. Fetal lung compression was suspected because the lung-thorax transverse area ratio was 0.13. The preload index of the inferior vena cava was 0.74, suggesting fetal cardiac failure. After fetal pleural effusion was aspirated, lung compression developed. Cordocentesis was performed at 33 weeks of gestation, and the fetal karyotype was confirmed to be 46, XY from an umbilical blood cultivation. The patient underwent a cesarean section at 33 weeks of gestation due to severe uterine contraction after preterm PROM. The baby was a 3,840 g male with a distended abdomen. Apgar score at 1 minute was 1. A chest X-ray demonstrated respiratory distress syndrome. The baby was discharged on the 69th day after birth and he is now 2 years and 9 months old and healthy. PMID- 10379504 TI - Massive fetomaternal hemorrhage: how long should children with good evolution be controlled? A case report. AB - We report on a term infant with a severe fetomaternal hemorrhage that caused a serious anemia that was surmounted after several transfusions. After the initial complications, such as persistent pulmonary circulation, severe anemia and thrombocytopenia, the outcome was good. We discuss the importance of a long-term follow-up of affected children, as well as their mothers. No clear parameters for a real prognosis are available. A follow-up is needed in order to detect possible complications in neurological development. PMID- 10379505 TI - Congenital neonatal myotonic dystrophy with persistent pulmonary hypertension and coma: a difficult diagnosis. AB - The fulminant forms of congenital myotonic dystrophy, which rapidly lead to death, are difficult to diagnose. The case described illustrates the roles of persistent pulmonary hypertension in such a fatal form. PMID- 10379506 TI - The midline dose distribution for a three-field radiotherapy technique. AB - At the University of Florida, head and neck cancer often is irradiated using parallel opposed lateral fields (with inferior borders slanted superiorly) and an anterior low neck field. A common criticism is that overlap may occur at the match-line junction of the three fields, resulting in an increased risk of radiation myelitis. One setup for treatment of the oropharynx and two for the larynx were irradiated in an anthropomorphic head and neck phantom made of tissue equivalent polyacrylamide gel with a two-dimensional thermoluminescent dosimeter array in its sagittal midplane. The results showed that no excess radiation dose was measured at the junction of the three fields. The "spinal cord dose," as percentage of dose to the central axis of the primary field, was as follows: oropharynx setup, 15% to 100%; larynx setup with midline tracheal block, 10% to 90%; larynx setup without tracheal block, 10% to 90%. In conclusion, the University of Florida three-field technique for head and neck cancer produces no measured increase in dose at field junctions. PMID- 10379507 TI - The effect of beam divergence on target coverage. AB - The well-known fact that radiation beams diverge is frequently not considered during the treatment planning process. Complacency with respect to beam divergence can, in some situations, lead to inappropriate field design. In this review, the potential problems arising from failure to adequately account for beam divergence in treatment planning are outlined, and commonly encountered clinical examples are illustrated. PMID- 10379508 TI - Clinical efficacy of respiratory gated conformal radiation therapy. AB - One major limitation of three-dimensional conformal radiation therapy that has not been adequately addressed is respiration-induced organ motion. During respiration, tumors in the abdomen can typically move from 1 to 3 centimeters. Because the size and shape of external radiation treatment fields do not change during treatment, the field size of the x-ray beam must be enlarged to encompass the tumor through the entire respiration cycle. Several manufacturers are developing respiratory gating systems. These systems allow the selective delivery of absorbed doses to moving target volumes in the abdomen during time intervals when the target volume is within the intended location. Before respiratory gated radiotherapy can be implemented clinically, the efficacy of the procedure must be justified. The magnitude of dosimetric and geometric uncertainties associated with respiratory motion must be identified to determine if gating can provide an advantage over conventional treatment techniques. In addition, clinical situations and specific types of cancer that could benefit from respiratory gating must also be identified. PMID- 10379509 TI - Technical management of a pregnant patient undergoing radiation therapy to the head and neck. AB - The fetal dose in a pregnant patient undergoing radiation therapy to the head and neck region was investigated. Implicit in this study was the design and evaluation of a shield used to minimize the fetal dose. To evaluate the fetal dose, a phantom was irradiated with the fields designed for this patient's therapy. The peripheral dose was measured for each field individually, both without and with a custom shield designed to be placed about the patient's abdominal and pelvic regions. The total dose at the location of the fetus over the course of this patient's radiation therapy was then estimated from peripheral dose rate measurements made at several points within the simulated uterus. With no shielding, the total dose within the uterus of the patient would have ranged from 13.3 cGy at the cervix to 28 cGy at the fundus. With the shield applied, the uterine dose was significantly less: 3.3 cGy at the cervix to 8.6 cGy at the fundus. In fact, at every measurement point, the peripheral dose with the shield in place was 30% to 50% of the dose without the shield. Some data suggest that the rate of significant abnormalities induced by irradiation in utero increases with increasing dose within the range of total peripheral doses incurred during most radiation treatment courses. It is therefore prudent to make reasonable attempts at minimizing the dose to the lower abdominal and pelvic regions of any pregnant patient. The shield designed in this work accomplished this goal for this patient and is flexible enough to be used in the treatment of almost all tumor volumes. PMID- 10379510 TI - Trimmed radiosurgical fields. AB - Radiosurgery aims to deliver a high radiation dose to a small target volume while sparing surrounding healthy tissues. However, since the target volume is often large and irregularly-shaped, a significant amount of healthy tissue is irradiated. To improve conformity of the dose volume to the target volume, we propose to optimize the field shape by trimming the field described by the radiosurgery cone with the accelerator jaws for a given arc. We have measured output factors (OF), tissue-maximum ratios (TMR), off-axis ratios (OAR) and penumbrae for 40, 32.5 and 24 mm cone fields trimmed by the lower (i.e., X jaws) and /or upper (i.e., Y jaws) collimator jaws. The smallest field was 8 mm large, and length was limited by the cone size. The average penumbra due to the cone field is 2.8 mm, and 4.1 and 6.1 mm for those due to the X and Y jaws, respectively. Moreover, the penumbrae due to the X and Y jaws are independent of jaw position within the radiosurgical field. Because of the large penumbra involved with the Y jaws, radiosurgical fields should be trimmed by the X1 and/or X2 jaws only. The measured OF's have been fitted with a hyperbolic function. All of the fitted OF's fall within +/- 0.5% of the measured OF's. The TMR values obtained with trimmed fields do not change much, except for the smallest fields (up to 10% at a depth of 20 cm). Therefore, using trimmed radiosurgical fields requires straightforward dosimetric changes and provides a level of beam shaping for large cone fields (> 20 mm in diameter) without introducing additional hardware. PMID- 10379511 TI - Spinal cord dose is higher than expected in head and neck radiation. AB - Myelopathy is a feared consequence of radiation therapy. Risk factors are multifocal; therefore, total dose calculation is crucial. We evaluated the contribution of scatter radiation to obtain an accurate cumulative spinal cord dose. Twenty patients undergoing three field head and neck radiation by Cobalt or 6 MV Linac had a total cord dose calculated from direct and scatter radiation. The cord was removed from the radiation field at tumor doses no higher than 4,400 cGy. Total tumor dose ranged from 5,400-7,400 cGy (mean 6060). All patients achieved the prescribed dose and none were lost to follow up (mean 36 months). It was found that scatter radiation can contribute as much as 20% extra dose to the spinal cord. Mean extra dose was 9% (range 1%-20%). This additional dose ranged from 52-810 cGy (mean 339 cGy). No apparent difference was seen with Cobalt or Linac source. Our conclusion was that significant additional dose is delivered to the spinal cord by scatter radiation and that scatter may contribute more to the development of myelopathy than previously believed. PMID- 10379512 TI - Comparison of kilovoltage x-ray and electron beam dose distributions for radiotherapy of the sternum. AB - The dose distributions for a patient with cancer involving the sternum were calculated for both a kilovoltage x-ray beam and a megavoltage electron beam. The minimum target dose and dose uniformity over the target volume were significantly better using electrons (90%-101%) than kilovoltage x-rays (68%-119%). The calculated lung dose and integral patient dose were also less for electrons than kilovoltage x-rays. For treating cancers of the sternum with radical intent, megavoltage electrons are recommended as the treatment mode of choice rather than kilovoltage x-rays. PMID- 10379513 TI - Commissioning, clinical implementation and quality assurance of Siemen's Virtual Wedge. AB - This report presents the results of commissioning, clinical implementation and quality assurance of Siemens Virtual Wedge. Our measurements show that: (1) wedge factors are within 2% of unity, (2) percentage depth doses are within 1% of open beam data, and (3) wedged beam profiles can be modeled similarly to a physical wedge and follow a well defined equation to facilitate modeling of an arbitrary wedge angle. The gantry angle dependence of wedge profiles is similar to open beam profiles. The output of wedged fields is linear with delivered monitor units within 1%. Quality assurance results indicate the wedge profiles are very stable over time. Day to day variations of two points measured along the wedge gradient direction are within 1.5%. PMID- 10379514 TI - On the future of genetic research in bipolar and schizophrenic syndromes. PMID- 10379515 TI - Dopamine D4 receptor gene: novelty or nonsense? AB - Although the role of genetics in personality has been studied extensively at a phenomenological level, only lately has the investigation of specific genes been performed. Recent reports suggest that DNA variants of the dopamine D4 receptor gene (DRD4) are associated with the personality trait of novelty seeking; however, others fail to replicate this finding. Such conflicting results suggest either a weak effect, an association only in certain populations, or a false positive resulting from population stratification. We provide a critical analysis of genetic studies of DRD4 variants with novelty seeking, alcoholism, drug abuse, and attention deficit hyperactivity disorder. Evidence for the role of DRD4 in novelty seeking is inconclusive, with a number of methodological concerns. Use of more conservative statistical criteria for significance, employing gene haplotypes, as well as linkage disequilibrium studies, are recommended. The molecular biology of the D4 gene is also reviewed. PMID- 10379516 TI - Association of the MscI polymorphism of the dopamine D3 receptor gene with tardive dyskinesia in schizophrenia. AB - In 112 schizophrenic patients previously treated with typical neuroleptics, we investigated the putative role of the dopamine D3 receptor gene (DRD3) in tardive dyskinesia (TD). Patients were assessed for TD severity using the Abnormal Involuntary Movement Scale (AIMS) and were subsequently genotyped for the MscI polymorphism that identifies a serine to glycine substitution in DRD3. A modified analysis of covariance model, which incorporated several clinical risk factors for TD, was utilized to detect differences in TD severity among the various genotypic groups. The glycine allele of DRD3 was found to be associated with typical neuroleptic-induced TD (F[2,95] = 8.25, p < .0005). Higher mean AIMS scores were found in patients homozygous for the glycine variant of the DRD3 gene, as compared to both heterozygous and serine homozygous patients. Although replication is necessary, this finding supports a role for the dopamine D3 receptor in the pathogenesis of TD. PMID- 10379517 TI - Effects of sustained phencyclidine exposure on sensorimotor gating of startle in rats. AB - Phencyclidine (PCP), a non-competitive NMDA antagonist with actions at multiple other central nervous system receptors, can cause both acute and lasting psychoses in humans, and has also been used in cross-species models of psychosis. Acute exposure to PCP in rats produces behavioral changes, including a loss of prepulse inhibition (PPI) of the startle reflex, which parallels the loss of PPI observed in schizophrenia patients. Sustained exposure to PCP in rats produces neuropathological changes in several limbic regions and prolonged behavioral abnormalities that may parallel neuropsychological deficits in schizophrenia. It is unclear whether sustained PCP exposure will also produce a loss of prepulse inhibition which parallels the decrease observed in schizophrenia patients. In the present study, we examined changes in PPI during and after sustained PCP administration, using 5-day PCP exposure via subcutaneous osmotic minipumps, or 14-day PCP exposure via repeated intraperitoneal injections. In both forms of drug delivery, PPI was disrupted during, but not after, sustained drug exposure. PPI does not appear to be sensitive to neuropathological effects of sustained PCP exposure. PMID- 10379518 TI - Neuroendocrine and psychophysiologic responses in PTSD: a symptom provocation study. AB - Biological research on post-traumatic stress disorder (PTSD) has focused on autonomic, sympatho-adrenal, and hypothalamo-pituitary-adrenal (HPA) axis systems. Interactions among these response modalities have not been well studied and may be illuminating. We examined subjective, autonomic, adrenergic, and HPA axis responses in a trauma-cue paradigm and explored the hypothesis that the ability of linked stress-response systems to mount integrated responses to environmental threat would produce strong correlations across systems. Seventeen veterans with PTSD, 11 veteran controls without PTSD, and 14 nonveteran controls were exposed to white noise and combat sounds on separate days. Subjective distress, heart rate, skin conductance, plasma catecholamines, ACTH, and cortisol, at baseline and in response to the auditory stimuli, were analyzed for group differences and for patterns of interrelationships. PTSD patients exhibited higher skin conductance, heart rate, plasma cortisol, and catecholamines at baseline, and exaggerated responses to combat sounds in skin conductance, heart rate, plasma epinephrine, and norepinephrine, but not ACTH. The control groups did not differ on any measure. In canonical correlation analyses, no significant correlations were found between response systems. Thus, PTSD patients showed heightened responsivity to trauma-related cues in some, but not all, response modalities. The data did not support the integrated, multisystem stress response in PTSD that had been hypothesized. Individual response differences or differing pathophysiological processes may determine which neurobiological system is affected in any given patient. PMID- 10379519 TI - Subtypes of family history and conduct disorder: effects on P300 during the stroop test. AB - The goal of the present study was to identify neurophysiological differences associated with a family history of substance dependence, and its subtypes (paternal alcohol, cocaine, or opiate dependence), and with conduct disorder, and its subtypes (aggression, deceitfulness/theft, and rules violations). P300 event related brain potentials were recorded from 210 males and females, aged 15-20 years while they performed the Stroop color-word compatibility test. Analyses revealed no significant effects of familial substance dependence on P300. However, an elevated number of conduct disorder problems was associated with a statistically significant reduction in P300 amplitude. The P300 amplitude reduction was related to the severity of the "rules violation" subtype, but was unrelated to aggression or deceitfulness and theft. It is concluded that conduct disorder can explain many of the P300 findings previously attributed to a family history of alcohol dependence. Furthermore, it appears that conduct disorder may be a heterogenous classification comprised of neurophysiologically different subtypes. PMID- 10379520 TI - MRI analysis of the cerebellum in bipolar disorder: a pilot study. AB - Since qualitative CT studies have suggested decreased cerebellar size in patients with bipolar disorder, we performed a quantitative analysis of the cerebellum in patients with bipolar disorder to determine whether high-resolution, thin slice magnetic resonance imaging (MRI) morphometry would reveal similar results. Bipolar patients hospitalized for a first manic episode (n = 16), bipolar patients with prior manic episodes hospitalized for a manic episode (n = 14), and normal volunteers (n = 15) matched for age, sex, race, and education were recruited and anatomic brain scans were acquired using a Picker 1.5 Tesla MRI scanner. Right and left cerebellar hemisphere volumes and vermal areas V1 (lobules I-V), V2 (lobules VI-VII), and V3 (lobules VIII-X) were measured. ANCOVA comparing each ROI, adjusting for race, sex, age, total cerebral volume, and substance abuse duration, revealed a significant group effect for vermal V3 area. Specifically, V3 area was significantly smaller in multiple-episode patients than in first-episode patients or healthy volunteers. Number of previous episodes of depression may contribute to this finding. These results suggest that cerebellar vermal atrophy may be a later neurodegenerative event in patients with bipolar disorder who have had multiple affective episodes. The confounding effects of medications are considered. PMID- 10379522 TI - Atrophic and static (neurodevelopmental) schizophrenic psychoses: premorbid functioning, symptoms and neuroleptic response. AB - The question of whether schizophrenic-like disorders are neurodevelopmental or degenerative in origin has been argued since the time of Kraepelin. The authors provide evidence for the existence of two etiologically distinct endophenotypes of the psychoses contained within the rubric of familial non-affective psychosis (schizophrenia), one atrophic and the other neurodevelopmental. The atrophic psychosis, identified by progressive ventricular enlargement throughout adult illness, evidences progressive impairment of interests, relationships, and withdrawal from latency through adolescence, with emergence of trait-like negative symptoms which are only marginally responsive to conventional neuroleptics. This psychosis also exhibits delayed response of positive symptoms during neuroleptic treatment, and may also proceed to a praecox dementia in later life. In contrast, a putative neurodevelopmental psychosis, associated with static ventricles during the course of adult illness, also demonstrates preadolescent impairments, but impairments which do not progress to marked negative symptoms. Conventional neuroleptics appear to have little effect (except sedation) on positive symptoms, but appear to induce negative symptomatology and partial disengagement from the burden of persistent psychotic thought processes in such static ventricle psychoses. Thus, separate patterns of illnesses with different prodromal features, different treatment response patterns, and different patterns of residual (negative) symptoms appear to characterize patients with psychosis who have expanding as opposed to stable cerebral ventricles at doses of neuroleptic at 10 mg haloperidol equivalents/day. PMID- 10379521 TI - Brain noradrenergic receptors in major depression and schizophrenia. AB - The binding of [125I]p-iodoclonidine to alpha-2, and/or [125I]iodopindolol to beta-1 and beta-2 adrenoceptors was measured in right prefrontal cortex (Brodmann's area 10) and right hippocampus from subjects with DSM-III-R diagnoses of major depression (n = 15) or schizophrenia (n = 8) as well as from control subjects (n = 20). No significant differences between study groups were observed in binding to alpha-2 adrenoceptors in any of the six layers of prefrontal cortex or in any of the hippocampal fields. Likewise, there were no significant differences in beta-1 or beta-2 adrenoceptor binding in any of the hippocampal fields between control and major depressive subjects. In contrast, binding to beta-1 adrenoceptors, but not beta-2 adrenoceptors, was significantly lower (-13 to -27%) in most hippocampal fields of schizophrenic subjects as compared to control subjects or to major depressives. Alterations in beta-1 adrenoceptor binding in the hippocampus of schizophrenics provide further evidence for a role of central noradrenergic neurons in the neurochemical pathology of schizophrenia. PMID- 10379523 TI - Differential induction of Fos-like-immunoreactivity in the extended amygdala after haloperidol and clozapine. AB - The extended amygdala is composed of the central and medial amygdaloid nucleus which through the sublenticular extended amygdala (SLEA) and the interstitial nucleus of the posterior limb of the anterior commissure (IPAC) merge into the bed nucleus of stria terminals (BST). Based on anatomical connections with limbic areas, the extended amygdala has been proposed to play an important role in cognitive and affective processes. This study examines the effect of the atypical antipsychotic clozapine and the classical antipsychotic haloperidol on Fos-like immunoreactivity (FLI) induction in areas belonging to the extended amygdala. Acute administration of clozapine (10-20 mg/kg) induced FLI in the central amygdaloid nucleus, IPAC, SLEA, and BST lateral division and, as previously described, in areas connected to the extended amygdala, such as the prefrontal cortex and nucleus accumbens shell. In contrast, acute administration of haloperidol (0.1-1 mg/kg) failed to induce FLI in the BST lateral division and SLEA but increased FLI in the IPAC. A small increase in FLI was observed in the central amygdaloid nucleus after 0.1 but not after 1 mg/kg of haloperidol. The present results, showing a preferential influence of clozapine, as compared to haloperidol, in the extended amygdala propose a new brain structure involved in the pharmacological effects of atypical antipsychotics. PMID- 10379524 TI - Raclopride and chlorpromazine, but not clozapine, increase muscle rigidity in the rat: relationship with D2 dopamine receptor occupancy. AB - The aim of the present study was to investigate the relationship between effects on muscle tone and D2 receptor occupancy of two typical antipsychotic drugs, raclopride and chlorpromazine, and the atypical drug, clozapine. Increased muscle tone (i.e., muscle rigidity), was measured as increases in tonic electromyographic (EMG) activity of the antagonistic muscles of the rat hind limb. D2 dopamine receptor occupancy was assessed in the striatum and substantia nigra, areas involved in the regulation of muscle tone. Raclopride and chlorpromazine produced dose-dependent increases in EMG activity associated with D2 occupancy of 68%-80% in the striatum and 67%-76% in the nigra. No significant increases in EMG were observed with clozapine which showed low D2 occupancy. The results are consistent with those from human studies showing extrapyramidal side effects were associated with striatal D2 occupancy of > 70%. PMID- 10379525 TI - Neurovascular deficits in cocaine abusers. AB - The nature of the neurological and cerebrovascular deficits in cocaine abusers and whether they persist in abstinence is unclear. Blood flow velocity of the anterior and middle cerebral arteries was measured by transcranial Doppler sonography in cocaine abusers (n = 50) and control subjects (n = 25). Blood flow velocity was measured within 3 days and again after about 28 days after being admitted to an inpatient research ward to determine whether blood flow velocity improved during monitored abstinence conditions. The mean, systolic, and diastolic velocities as well as the pulsatility index in middle and anterior cerebral arteries significantly differed between controls and cocaine abusers (p < .05). Cerebrovascular resistance is increased in cocaine abusers and the increase persists for over a month of abstinence. Further research is needed to determine whether cerebrovascular resistance can be improved by pharmacological manipulations and whether improved blood flow relates to improved treatment outcome. PMID- 10379526 TI - Noribogaine generalization to the ibogaine stimulus: correlation with noribogaine concentration in rat brain. AB - The discriminative stimulus effects of ibogaine and noribogaine in rats have been examined in relation to their concentrations in blood plasma and brain regions and to receptor systems through which they have been proposed to act. Rats were trained to discriminate ibogaine (10 mg/kg i.p.), the NMDA antagonist dizocilpine (0.08 mg/kg i.p.) or the kappa-opioid agonist U50,488 (5 mg/kg i.p.) from vehicle in a standard two-lever operant conditioning procedure with a tandem VI-FR schedule of food reinforcement. Only rats trained on ibogaine generalized to noribogaine, which was approximately twice as potent as the parent compound. Noribogaine was detected in plasma and brain after administration of ibogaine and noribogaine. At the ED50 doses for the discriminative effect, the estimated concentrations of noribogaine in plasma, cerebral cortex, and striatum were similar regardless of whether ibogaine or noribogaine was administered. The findings suggest that the metabolite noribogaine may be devoid of NMDA antagonist and kappa-opioid agonist discriminative effects and that it may play a major role in mediating the discriminative stimulus effect of ibogaine. PMID- 10379527 TI - Altered activity of midbrain dopamine neurons following 7-day withdrawal from chronic cocaine abuse is normalized by D2 receptor stimulation during the early withdrawal phase. AB - Using in vivo single-unit recording in rats, we compared the effects of continuous cocaine infusion via minipump or single daily injections (both 40 mg/kg/d x 14 days, S.C.) on the activity of putative dopamine (DA) neurons in the substantia nigra pars compacta (SNC) and ventral tegmental area (VTA). On days 1 5 after cocaine withdrawal, animals were further treated with single daily injections of DA agonists. On withdrawal day 7 continuous cocaine caused a reduction in spontaneously active neurons in the SNC and reduced bursting in the VTA. In contrast, intermittent cocaine resulted in an increase in the number of active neurons in the VTA. These changes were all reversed by apomorphine or quinpirole given during the first 5 withdrawal days. The D1 antagonist SCH 39166 did not antagonize the effects of apomorphine in either region. The role of D2 receptors in modulating baseline DA activity during intermediate cocaine withdrawal is discussed. PMID- 10379528 TI - Excitatory actions of NMDA receptor antagonists in rat entorhinal cortex and cultured entorhinal cortical neurons. AB - We have characterized excitatory effects of non-competitive NMDA receptor antagonists MK-801, PCP, and ketamine in the rat entorhinal cortex and in cultured primary entorhinal cortical neurons using expression of immediate early gene c-fos as an indicator. NMDA receptor antagonists produced a strong and dose dependent increase in c-fos mRNA and protein expression confined to neurons in the layer III of the caudal entorhinal cortex. Induction of c-fos mRNA is delayed and it is inhibited by antipsychotic drugs. Cultured entorhinal neurons are killed by high doses of MK-801 and PCP but c-fos expression is not induced in these neurons indicating that this in vitro model does not fully replicate the in vivo effects of PCP-like drugs in the entorhinal cortex. Excitatory effects of the NMDA receptor antagonists may be connected with the psychotropic side effects of these drugs and might become a useful model system to investigate neurobiology of psychosis. PMID- 10379530 TI - The validity of the PET/alpha-[11C]methyl-L-tryptophan method for measuring rates of serotonin synthesis in the human brain. PMID- 10379529 TI - Psychological states and lymphocyte beta-adrenergic receptor responsiveness. AB - There is a complex interplay between psychological states and biochemical factors. beta-Adrenergic receptor responsiveness is altered in some patients with depression and anxiety disorders, but the relation between various psychological states and receptor function in a normal population is unknown. We measured lymphocyte beta-adrenergic receptor density (Bmax), sensitivity (cAMP ratio), the Profile of Mood States (POMS), and Spielberger State-Trait Anxiety Inventory (STAI) in 39 hypertensives and 81 normotensives. We examined correlations between log normalized receptor variables and psychological states. Log Bmax showed negative correlations with age and with POMS tension-anxiety, depression dejection, and anger-hostility. Log cAMP ratio did not show significant correlations with POMS and STAI ratings. In step-wise multiple regression analyses, 36% of the variance in Bmax was accounted for by POMS tension-anxiety, and age. Our study suggests that increased POMS tension-anxiety was highly associated with down-regulation of beta-adrenergic receptors, even in subjects who do not have psychiatric illness. Numerous psychological states could be associated with changes of beta-adrenergic receptor responsiveness in a normal population. PMID- 10379531 TI - [Barrett's esophageal cancer]. AB - Barrett's esophageal cancer is generally considered to be generated through the metaplasia-dysplasia-carcinoma sequence from Barrett's epithelium. We have examined 9 cases of Barrett's esophageal cancer resected in our department, and determined that: 1) The cancer is mainly located in the lower esophagus. 2) Metastases are commonly found in the abdomen and mediastinum in advanced-stage disease. 3) Long-term survival can be expected in patients who undergo radical surgery. PMID- 10379532 TI - [Molecular alterations in Barrett's esophagus and adenocarcinoma]. AB - Patients with Barrett's columnar-lined esophagus are at increased risk of developing esophageal adenocarcinoma, the incidence of which has increased rapidly especially in the USA. Although the number of patients with Barrett's adenocarcinoma is fewer in Japan than in the USA, all gastroenterologist should know its multistep carcinogenic process. Tumor suppressor genes (p53, p16), oncogenes (c-erbB-2, H-ras, K-ras, cyclin D1, src), and growth factor/receptor (TGF-alpha, EGFR) seem to cause the malignant transformation of Barrett's esophagus. Because detection of these molecular alterations is feasible, more accurate diagnosis of Barrett's esophageal biopsy specimens should be made by adding the molecular examination to the conventional pathologic examination. PMID- 10379533 TI - [Histologic occurrence of Barrett's carcinoma]. AB - It is generally agreed that the diagnosis of Barrett's esophagus is justified when the columnar epithelium 3 cm or more from the E-C junction is involved. Microscopically, the epithelium has three distinct appearances: (1) gastric fundic type; (2) junctional type: and (3) specialized columnar type. In mucin histochemistry in 6 cases of Barrett's carcinoma, 5 cases of Barrett's esophagus were positive for HID-AB. Four cases of Barrett's carcinoma were also positive. These results suggest that Barrett's carcinoma developed from the specialized columnar type of Barrett's esophagus. PMID- 10379534 TI - [Endoscopic diagnosis of Barrett's adenocarcinoma]. AB - Biopsy specimens can reveal that esophageal cancer is an adenocarcinoma but they cannot show that its origin is Barrett's mucosa. Therefore we must show during endoscopy that the tumor exists in Barrett's mucosa. We reported that Barrett's esophagus could be clearly diagnosed at endoscopy as the columnar mucosa lying on the longitudinal vessels in the lower esophagus. We define Barrett's esophagus as "the columnar mucosa in the esophagus which exists continuously more than 2 cm in circumference from the stomach." Short-segment Barrett's esophagus (SSBE) is "the columnar mucosa which exists in the esophagus continuously from the stomach but its length has a part under 2 cm in length." Endoscopically Barrett's adenocarcinoma is visualized as a lesion with a reddish and uneven mucosal surface. Barrett's adenocarcinomas occur in the SSBE as well. Endoscopic observation at periodic intervals is necessary not only for cases with Barrett's esophagus but also with SSBE. A further examination is necessary to determine the application of EMR for superficial Barrett's adenocarcinoma. PMID- 10379535 TI - [Treatment of Barrett's esophageal cancer with special reference to endoscopic treatment]. AB - Most cases of Barrett's esophageal cancer are discovered in the advanced stage, and therefore the principle treatment is surgical. Endoscopic treatment is employed for patients with early cancer and severe dysplasia discovered by endoscopic examination, accidentally or during follow-up of Barrett's esophagus. Among the endoscopic forms of treatment, photodynamic therapy (PDT) is the main procedure for early cancer and severe dysplasia in Barrett's esophagus. Recently argon plasma coagulation (APC) has been introduced for the treatment of severe dysplasia and early cancer, in addition to Barrett's epithelium itself. In Japan, endoscopic mucosal resection (EMR) has also been employed in 5 cases of mucosal cancer in Barrett's esophagus. It will be used more widely in Japan, because the number of reflux esophagitis and Barrett's esophagus cases will increase in the near future. PMID- 10379536 TI - [Surgical treatment of adenocarcinoma in Barrett's esophagus and prognosis]. AB - There is no consensus regarding the surgical approach to adenocarcinoma in Barrett's esophagus. From 1980 to 1988, 8 patients with adenocarcinoma in Barrett's esophagus were treated at the National Cancer Center Hospital. Seven patients underwent subtotal esophagectomy with extended lymph node dissection, and one transhiatal esophagogastrectomy with regional lymph node dissection. In 4 patients tumor invasion was limited within the submucosa and in 4 within the muscularis propria. Four of 8 patients had stage I disease. The 5-year survival rate for the 8 patients was 64.3%. Some reports have indicated that endoscopic survey for Barrett's esophagus is important for early diagnosis. We conclude that survival after esophagectomy for adenocarcinoma in Barrett's esophagus is dependent on the method of operation, and that patients with early lesions may expect significantly better survival after extended lymph node dissection. PMID- 10379537 TI - [Therapeutic strategy for adenocarcinoma in Barrett's esophagus: a study based on a comparison with squamous cell carcinoma]. AB - The therapeutic strategy for adenocarcinoma in Barrett's esophagus is discussed based on a comparison with squamous cell carcinoma. The pattern and range of lymph node metastasis of adenocarcinoma in Barrett's esophagus is similar to that of squamous cell carcinoma, as is the pattern of recurrence. Chemotherapy is less effective, except for some reports on paclitaxel, but chemoradiation therapy is comparable in effect. The general tendency of a better prognosis for patients with carcinoma in Barrett's esophagus is the result of several factors, such as earlier detection of the disease, lower probability of lymph node metastasis, and distal location which permits less radical curative surgical procedures without cervical and superior mediastinal lymph node dissection. In principal the therapeutic strategy for adenocarcinoma in Barrett's esophagus should be determined like that for squamous cell carcinoma, taking into account the location of the lesion and the depth of invasion. PMID- 10379539 TI - Conditions for coherent vibrations in the cytoskeleton. AB - Mechanism of organization and order in biological systems is not satisfactorily explained yet. Biophysical theories predicted that coherent endogenous electric field of high frequency can play a significant role in organization. The polarity of vibration structures, spectral energy transfer caused by nonlinearities, and energy supply can lead to energy condensation and excitation of coherent states. These conditions are satisfied in the polymer structures of the cytoskeleton, in particular, in the microtubules as follows from experimental findings. Nonetheless, experimental verification of energy condensation and coherent vibrations in the cytoskeleton is still missing. PMID- 10379538 TI - Microelectronic sensors for measurement of electromagnetic fields of living cells and experimental results. AB - Microelectronic sensors are used for measurements of electromagnetic fields generated by synchronized cultures of yeast cells. Cold sensitive mutant tub2-401 of Saccharomyces cerevisiae is used. The measured electromagnetic signals in the frequency range from 8 to 9 MHz are compared with evolution of the reassembled microtubules. The detected signals peak in the time interval 25-30 min and 45-60 min after the release of the cells from the restrictive to the permissive temperature. The first maximum corresponds to the stage when the mitotic spindle is formed and binds chromatids. The second maximum is measured when the processes of anaphase A and of anaphase B take place. PMID- 10379540 TI - Quantum-mechanical coherence in cell microtubules: a realistic possibility? AB - We discuss the possibility of quantum-mechanical coherence in Cell MicroTubules (MT), based on recent developments in quantum physics. We focus on potential mechanisms for 'energy-loss-free' transport along the microtubules, which could be considered as realizations of Frohlich's ideas on the role of solitons for superconductivity and/or biological matter. In particular, by representing the MT arrangements as cavities, we review a novel scenario, suggested in collaboration with D.V. Nanopoulos, concerning the formation of macroscopic (or mesoscopic) quantum-coherent states, as a result of the (quantum-electromagnetic) interactions of the MT dimers with the surrounding molecules of the ordered water in the interior of the MT cylinders. We suggest specific experiments to test the above-conjectured quantum nature of the microtubular arrangements inside the cell. These experiments are similar in nature to those in atomic physics, used in the detection of the Rabi-Vacuum coupling between coherent cavity modes and atoms. Our conjecture is that a similar Rabi-Vacuum-splitting phenomenon occurs in the absorption (or emission) spectra of the MT dimers, which would constitute a manifestation of the dimer coupling with the coherent modes in the ordered water environment (dipole quanta), which emerge due to the phenomenon of 'super radiance'. PMID- 10379541 TI - Microtubules: strange polymers inside the cell. AB - This paper provides a consistent approach (within a one-dimensional approximation) to the description of the evolution of the microtubule length at both low- and high-density concentrations. We derive general master-type equations which are based on the key chemical reactions involved in the assembly and disassembly of microtubules. The processes included are: polymerization and depolymerization of a single protein dimer, catastrophic disassembly affecting an a piori arbitrary number of dimers, and a rescue event. Solutions of the derived equations are compared with the existing experimental data. Important conclusions linking the emergence of bell-shaped histograms with the nature of catastrophe and rescue phenomena are drawn. Finally, we briefly discuss the emergence of coherent phenomena in microtubule polymerization, i.e., a transition to collective oscillations in the assembly and disassembly effects. PMID- 10379542 TI - Damping and modification of the multiquanta Davydov-like solitons in molecular chains. AB - Relaxation of the multivibron soliton in molecular chain on lattice vibrations is investigated within the simple microscopic model. It was shown that its dynamics is governed by the nonlinear Schrodinger equation containing damping term. Its appearance is the consequence of the emission and absorption of real phonons arising when soliton velocity approaches the phase speed of sound. Explicit time dependence of the soliton parameters results as a consequence of these perturbations. In particular, soliton amplitude decreases while its width increases. PMID- 10379543 TI - Elementary arguments that the Wu-Austin Hamiltonian has no finite ground state (the search for a microscopic foundation of Frolichs theory). AB - The Wu-Austin Hamiltonian as the basis for deriving Frohlichs rate equations from a microscopical point of view has been investigated. In addition to an earlier paper we show in a very easy manner that this or similar Hamiltonians have no lower bound and are therefore unphysical. The perturbation expansion which is the tool to derive Frohlichs rate equations with this Hamiltonian is not converging. Therefore, the usual derivation of this rate equation is not valid. PMID- 10379544 TI - Technical aspects of microwave thermotherapy. AB - We describe our new technical results dealing with microwave thermotherapy (hyperthermia) in cancer treatment, see Refs. [S.B. Field, C. Franconi (Eds.), Physics and technology of hyperthermia, NATO Seminar Proceedings, Urbino, Italy, 1986; J. Hand, J.R. James (Eds.), Physical Techniques in Clinical Hyperthermia, Wiley, New York, 1986; J. Vrba, M. Lapes, Microwave Applicators for Medical Purposes, CTU Press, 1996, in Czech; J. Vrba, C. Franconi, M. Lapes, Theoretical limits for the penetration depth of the intracavitary applicators, International Journal of Hyperthermia, 12:6 (1996) 737-742; C. Franconi, J. Vrba, F. Montecchia, 27 MHz hybrid evanescent-mode applicators with flexible heating field for deep and safe subcutaneous hyperthermia, International Journal of Hyperthermia, 9:5 (1993) 655-674.]. Our research interest is to develop applicators for deep local heating and for intracavitary cancer and/or prostate treatment as well. Further, a system for 3D SAR distribution measurements in water phantom is explained. Basic evaluation of clinical results is given. PMID- 10379545 TI - Electromagnetic-field-induced oscillations of the lipid domain structures in the mixed membranes. AB - The effect of external electromagnetic field (EMF) on a percolation structure formed during phase separation in the mixed phospholipid membranes was studied by computer simulation. Decay of the percolation structure under electromagnetic radiation was detected. It was shown that oscillation regime can be realized in this system: periodic alternation of formation and decay of the percolation cluster was observed under 10 kHz EMF. The decay of the lipid domain structure in the EMF results from anomalous increase of the permittivity of the continuous fluid lipid phase in the percolation threshold region. It is proposed that detected EMF effect can influence the signal and transport processes associated with percolation properties of biomembranes. PMID- 10379546 TI - Effects of sinusoidal magnetic field on adherence inhibition of leucocytes: preliminary results. AB - The leucocyte surface properties manifest the cell-mediated immunity. The response of the cell-mediated immunity to external magnetic field was examined by observing leucocyte adherence to solid state surfaces. In the presence of antigen, leucocytes taken from cancer patients exhibit decreased adherence in contrast with adherence of leucocytes from healthy humans. After 1 h exposure to a sinusoidal magnetic field of 50 Hz and of 1 mT or 10 mT, adherence of leucocytes taken from cancer patients is strongly increased. The 1 mT magnetic field has stronger effect than the 10 mT field. PMID- 10379547 TI - On the possible role of the nitrogen atom in living matter. AB - Molecules of NH3 are capable of emitting stimulated radiation (MASER). Their organic derivates could have a similar effect, as is suggested by the new results published in Electron Correlations in Molecules and Solids [P. Fulde, Electron Correlations in Molecules and Solids, Springer-Verlag, Berlin, Heidelberg, 1993]. PMID- 10379548 TI - Nonlinear dynamics of a DNA chain affected by endogenous AC fields. AB - In this paper we investigate the nonlinear dynamics of a DNA chain in the presence of endogenous AC fields (EACF) generated by the living cell itself. The dynamics of the DNA chain is described in the framework of the nonlinear breather mode. The transition of breather localized modes into open states, affected by AC fields is calculated by using Kubo's formalism for the linear response of the system. PMID- 10379549 TI - Peculiarities of long-range interaction between the nucleotides after DNA damage. AB - In this work, the long-range interaction between the pairs of nucleotides situated on the opposite ends of a double broken DNA helix have been studied theoretically. The long-range energy was considered as a sum of electrodynamics Van der Waals and electrostatic Coulomb interactions. The most important region for the problem under consideration is about 5-15 A between the nucleotides. The calculations of the energy of long-range interaction have shown that during the interaction between the pairs CG-CG, there is a potential repulsive barrier with the amplitude of about 4 kT at the distance of 7-9 A, and during the interaction between the pairs TA-TA, there is a potential repulsive barrier with the amplitude of about 1.5 kT at the same distance, which can prevent DNA from enzyme selfrepairing after a double DNA break. This barrier vanishes as the pH of intracellular medium increases. The remainder pairs of nucleotides do not have such barrier, and there is always an attraction like interaction. PMID- 10379550 TI - Influence of a periodic field on the distant electron transfer in biological systems. AB - Generalization of the Marcus transfer rate is derived for the case of a dissipative long-range donor-acceptor electron transfer (ET) mediated by specific bridging electron pathways in biological systems and driven by ac-electric field. High-frequency electric field is shown to block and even to invert the transfer if a specific relation between amplitude and frequency of the ac-field is fulfilled. PMID- 10379551 TI - Quantum dissipation and neural net dynamics. AB - Inspired by the dissipative quantum model of brain, we model the states of neural nets in terms of collective modes by the help of the formalism of Quantum Field Theory. We exhibit an explicit neural net model which allows to memorize a sequence of several informations without reciprocal destructive interference, namely we solve the overprinting problem in such a way last registered information does not destroy the ones previously registered. Moreover, the net is able to recall not only the last registered information in the sequence, but also anyone of those previously registered. PMID- 10379552 TI - Common model of evolution for living cell and central nervous system. AB - Both living cell and central nervous system can be treated as objects similar to artificial neural networks, actively studied now. One can see deep analogies between evolutionary processes in these systems, and correspondences can be established between some phenomena and objects. These are: genome vs. memory, gene vs. symbol, cell type vs. image, mitosis vs. sleep, organism vs. perception state, species vs. language, fertilization vs. attention, meiosis vs. paradoxal sleep. There is reason to study these correspondences on more technical level being based upon network nature of the living cell and nervous system. PMID- 10379553 TI - Reasons of poor replicability of nonthermal bioeffects by millimeter waves. AB - It is shown that the reasons for the poor replicability of the nonthermal bioeffects of radiofrequency have the fundamental bases. The efficiency of the living system process is determined by the conversion of electromagnetic energy into Helmholtz free energy. The efficiency dependence on the absorbed power of electromagnetic radiation is quite different in the Wien region (visible light) and in the Rayleigh-Jeans region (radiofrequency). In the Wien region the endergonic and exergonic processes could never be confused. In the Rayleigh-Jeans region the endergonic and exergonic processes could be confused. It gives the poor replicability. PMID- 10379554 TI - Problems of weak electromagnetic field effects in cell biology. AB - Electrostimulations of cells by weak electric or electromagnetic LF and HF-fields are applied widely today; capacitively or inductively coupled, however, they are seldom applied for cell-free and membrane-free solutions of enzymes. First, the detection of a response of the cells ('electrical window') is a prerequisite for testing at least three parameters: frequency, amplitude and treatment time, besides reproducible biological conditions. The 'state-of-the-art' of this fast developing direction of bioelectrochemistry can be characterized in the following way: the results from several laboratories of (a) cell proliferation, (b) ion transport, (c) activation of several enzymes (Na,K-ATPase), (d) increase of certain protein concentrations (heat-shock protein hsp70) are more or less in agreement. Unfortunately, there are discrepancies between no less than 7 labs in the gene expression of c-myc, c-fos histone 2B, -actin, URA-3 and others, especially for low fields (< 0.05 mT), e.g., in HL60 cells! The reason why seems to be: (1) differences in the most suitable isolation procedure, (2) interferences in the case of too low magnetic flux and (3) too small ranges of parameters have been measured. Today, three open problems must be pointed out: (A) What is the physiological causality for specific 'electrical windows' and their positive or negative efficacy? (B) What are the biochemical targets for either magnetic or electric fields or both? (C) What is the influence of electrical and (or) thermal noise on field efficiency? PMID- 10379555 TI - Hydration of phosphatidyl serine multilayers and its modulation by conformational change induced by correlated electrostatic interaction. AB - Hydration of the various residues of phospholipids was inferred from the shift in the wave number of their vibration bands, obtained from the amplitudes of their positive and negative peaks in the difference spectra between those of the hydrated and the dry phospholipid multibilayers. The effect of aligned phospholipid layers on the orientation of their hydrating water molecules was inferred from the dichroic ratio of the OH stretching band, measured by polarized attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) with a germanium prism, as a function of the water-to-lipid ratio in the surface film. The results indicate that about seven water molecules are oriented by one phosphatidyl serine molecule in the surface film. About 8 to 11 additional water molecules contribute to the hydration of the polar residues as revealed by the effect on the difference spectra. The hydration appears to be cooperative. A water molecule that initiates hydration of a site facilitates access of additional water molecules, until the hydration of the whole site composed of many different interacting polar residues is completed. PMID- 10379556 TI - Effects of ELF capacitively coupled weak electric fields on metabolism of 6B1 cells. AB - In this study, we adopted several methods of MTT colorimetry, DAPI fluorimetry and ELISA to study the effects of extremely low frequency (ELF) capacitively coupled electric fields (EFs) on the metabolism of 6B1 cells. The result shows that 50 mV cm(-1) ELF EF (10-100 Hz) has no significant effect on proliferation, DNA synthesis and activity of succinate dehydrogenase of 6B1 cells, indicating that the effect of ELF (10-100 Hz) EF on the metabolism of 6B1 cells is not obvious. However, 50 mV cm(-1), 50 Hz EF significantly promotes the HBs-Ab (Hepatitis B surface antibody) secretion of 6B1 cells, implying that under this situation, EF has some distinctive effect on the outerface of 6B1 cell membrane. PMID- 10379557 TI - A model for the generation of low level chemiluminescence from microbiological growth media and its depletion by bacterial cells. AB - We present a model for the development of chemiluminescence (CL) in autoclaved liquid growth media, as they are used in microbiology for the culturing of microorganisms, or in other related Maillard systems. The model distinguishes between four different stages consisting of sugar fragmentation during heating, autooxidation of highly reducing fragmentation products, radical chain reactions leading to a peroxidation of the media, and finally the formation of excited states, energy transfer reactions and CL emission. The proposed model is also discussed in regard of a recently reported elimination of this CL in growing cultures of microorganisms and possible pathways for this interference are suggested. PMID- 10379558 TI - High magnetic field enhances stationary phase-specific transcription activity of Escherichia coli. AB - When Escherichia coli B was aerobically grown at 37 degrees C under inhomogeneous 5.2-6.1 Tesla (T) magnetic fields in the superconducting magnet biosystem (SBS), the cell number in the stationary phase after the growth had leveled off, was about 3 times higher than that under a geomagnetic field. When the E. coli defective in the rpoS gene which encodes a sigma factor, sigmaS of RNA polymerase and is specifically expressed in the stationary phase was cultivated at 37 degrees C in SBS, such enhancement of cell survival was significantly reduced. The E. coli cells carrying rpoS-lacZ fusion gene or other rpoS dependent genes fused with lacZ were grown, significant increase in the activity of beta galactosidase was observed in the stationary phase under high magnetic field. These data suggest that enhancement of the transcription activity in stationary phase is involved in the higher survival of the cells under magnetic field. PMID- 10379559 TI - Genetic programming as an analytical tool for non-linear dielectric spectroscopy. AB - By modelling the non-linear effects of membranous enzymes on an applied oscillating electromagnetic field using supervised multivariate analysis methods, Non-Linear Dielectric Spectroscopy (NLDS) has previously been shown to produce quantitative information that is indicative of the metabolic state of various organisms. The use of Genetic Programming (GP) for the multivariate analysis of NLDS data recorded from yeast fermentations is discussed, and GPs are compared with previous results using Partial Least Squares (PLS) and Artificial Neural Nets (NN). GP considerably outperforms these methods, both in terms of the precision of the predictions and their interpretability. PMID- 10379560 TI - Free energy of charge transfer and intraprotein electric field: method of calculation depends on the charge state of protein at a given structure. AB - Free energy of charge transfer presents a basic characteristic of reactions such as protonation, oxido-reduction and similar. Evaluation of this quantity requires calculation of charging energy. Proteins are structured dielectrics, and a consistent incorporation of their structure into calculation of intraprotein electric field results in expression for charging energy of an active group in protein, which is essentially different from that for a simple dielectric. An algorithm for semi-continuum calculation of relevant free energies is described. First of the two components of charging energy in protein, energy of the medium response to charge redistribution in reactants, should be always calculated as the charging energy by the charge redistribution using the static dielectric constant of protein. The second term is interaction energy of the charge redistribution with the 'frozen' electric field of the system before reaction. Charges of protein groups, at which the protein structure has been determined, are often different from those before reaction of charge transfer, so is the corresponding intraprotein field. The field is expressed through either both the optical and static dielectric constants of protein or only optical one depending on whether the charges of protein groups before reaction and upon structural analysis are the same or not. Proper allowance for difference in charges of reacting groups before reaction and upon structural analysis of protein is thermodynamically necessary and quantitatively important. The expression for activation free energy for charge transfer in proteins is derived in the form presenting explicitly an invariant contribution of protein structure. PMID- 10379562 TI - Detection of the DeltaF508 mutation in the CFTR gene by means of time-resolved fluorescence methods. AB - A rapid recognition in the base sequence of nucleic acids is an important prerequisite toward the diagnosis of genetic diseases and their carrier states. We have developed a hybridisation method in which a fluorescently labeled oligonucleotide is used to detect point mutations in a target by a simple fluorescence lifetime analysis of the emission of the fluorescent label. We applied this method to detect the deltaF508 mutation in the cystic fibrosis transmembrane conductance regulator (CFTR) gene in a model system and with biologically derived PCR product and discuss the potential generality of this method. PMID- 10379561 TI - Calcium channels, potassium channels and membrane potential of smooth muscle cells of human allantochorial placental vessels. AB - The membrane potential (Um), the main factor of the excitation-contraction coupling, of human allantochorial placental vascular smooth muscle cells (VSMCs) has been previously shown to depend on voltage-sensitive K+ channels. These channels were blocked by high external K+. To characterize other channels which regulated Um, various constrictor or/and vasodilators and channel blockers were used. Serotonin depolarized VSMCs, in normal medium, but induced a more marked depolarization in VSMCs predepolarized by high external K+. This depolarization was inhibited by nifedipine, a blocker of voltage-gated Ca2+ channels. Acetylcholine, sodium nitroprusside (without effect on Um in normal medium), hyperpolarized the predepolarized-high K+ medium VSMCs. This hyperpolarization was inhibited after addition of charybotoxin (a blocker of Ca2+-activated K+ channels) or/and glibenclamide (a blocker of ATP-sensitive K+ channels). A similar effect was obtained with isoproterenol. These results indicated that membrane potential of human placental allantochorial VSMCs was regulated by voltage-gated, Ca2+- and ATP-sensitive K+ channels and by voltage-dependent Ca2+ channels. PMID- 10379563 TI - Theoretical aspects of single pulse induced modulation of cell-cell interactions favoring iso-electrofusion. AB - A single pulse-mediated electrofusion of freely suspending widely separated cells could reduce steps in the protocol. Prediction of such electrofusion is still unsolved. In pre-pulse condition, quantitative estimation with three-layered model cell surface reveals electrostatic repulsion (due to negative surface charge) is the major hindrance to membrane-to-membrane contact. On theoretical grounds we predict that designed single high voltage pulsing on widely separated model cell surface can counteract the non-specific repulsive interaction and favor approach of two apposed membranes. But hydrodynamic modulation of pulse exposed cell surface interaction can hamper approach of membranes contact when cells are held at a gap of more than approximately 450 A. PMID- 10379564 TI - Multiple-pulse-mediated electrofusion of intact erythrocyte onto human term placental amnion. AB - The creation of surface modified human term placental amnion by electrofusing human cells onto its surface has been thought of. A multiple-pulse electrofusion protocol with 10 square pulses of 10-micros pulse length, and electric field of 0.2 kV cm(-1), can make erythrocyte-amnion tissue electrofusion possible. The protocol devised merge the cell-tissue-adherence steps with fusogenic pulse. The finding opens up a new avenue of cell electrofusion onto human tissue with minimal procedural complexities. PMID- 10379565 TI - Membrane microextension: a possible mechanism for establishing molecular contact in electrofusion. AB - True cell membrane contact is an essential condition for electro-pulsed cell fusion, but initial morphological perturbation leading to true contact is still not clear. Dielectrophoresis mediated compression and fusogenic pulse induced compaction of cells led to rapid merger of tight membranes, and deprived direct microscopic view of surface membrane perturbation. Freely suspending cells with large and different cell-cell gaps may proceed to electrofusion with perturbed membrane and initiates fusion events at different time. These pulsed exposed cells can be used for capturing changes in the membrane surface and early electrofusion events. Early stage of fusion of freely suspended intact human erythrocytes exposed to single exponential decay pulse was studied by scanning electron microscopy (SEM). Field pulse induces small membrane bumps. Interaction of bumps on adjacent membranes lead to true membrane contact and form bridges between the membranes as microextension, combining both membranes into a topologically single structure. Some fusion products showed expanded fusion zones, which suggest indication of open lumen at contact area. PMID- 10379566 TI - Pore resealing inactivation in electroporated erythrocyte membrane irradiated with electrons. AB - The changes in the electroporation process of human erythrocytes membrane due to the direct action of high energy electron radiation were investigated. To avoid the indirect effects caused by radiolytic products of water, the irradiation was performed at liquid nitrogen temperature. The irradiated cells have been exposed to square-wave electric pulses at 4 degrees C in isotonic suspensions to induce membrane electropores. The pores resealing were quantified by monitoring the cell hemolysis. A significant decrease of the resealing process was found for irradiation doses higher than 100 Gy. The mass of molecular structures affected by the direct action of radiation was estimated using the target analysis method. We found a molecular weight Mm approximately 930 kDa roughly corresponding to spectrin tetramer of the cytoskeleton. This suggests that spectrin network plays an important role in the pores resealing of the electropermeabilized erythrocyte membrane. PMID- 10379567 TI - Influence of additives on the protoplasts electrofusion. AB - Various neutral or charged surface active substances were used for testing the influence of additives on the electrofusion of barley protoplasts. It was found that neutral surface active agents DX, TAGB, Span-80 and AEO-9 could promote the electrofusion. The positively charged surface active agents Bardac 2080, Bardac 2280 and amphoteric surface active agents dodecyl-propyl betaine and CAB betaine also promote the electrofusion, but at high concentration the electrofusion efficiency will reduce. The negatively charged polymer agents Cibacron blue DX, Fluoresceinylthiocarbamoyl DX, and active surface substances K12 and Carsonol TLS presented negative effect. These phenomena were discussed from the view of adsorption of additives on the membrane and the interactions between protoplasts. PMID- 10379568 TI - Electroporation-mediated topical delivery of vitamin C for cosmetic applications. AB - It is now medically recognized that sagging skin and other signs of degenerative skin conditions, such as wrinkles and age spots, are caused primarily by oxy radical damage. Vitamin C (Vit. C), in the form of L-ascorbic acid (Asc), is the one vitamin that can accelerate wound healing, protect fatty tissues from oxidation damage, and play an integral role in collagen synthesis. It is known that the lipid-rich stratum corneum (SC) is a highly resistant barrier to chemical agents penetrating into the skin. This report describes the first feasibility study of electroporation-mediated topical delivery (EMTD) of Asc for potential cosmetic applications. Both a cream formulation (20% Asc) and a crystal suspension (33% Asc) were applied respectively to human cadaver skin and fresh surgical skin. Six exponential pulses at 60 or 100 V and pulse lengths of 2.7-30 ms were selected. EMTD was more effective on fresh human skin than on human cadaver skin. For both skin models, EMTD with cream resulted in a greater enhancement of Vit. C penetration than with suspension. The distribution of electrical fields through the SC, epidermis, and dermis is demonstrated in computer simulation. Assuming that this fresh skin model and certain experimental conditions simulate projected in vivo applications, EMTD of Vit. C may represent an alternative method to ameliorate skin aging. PMID- 10379569 TI - Interactions between carbonic anhydrase and some decarboxylating enzymes as studied by a new bioelectrochemical approach. AB - This work presents the results of a study, carried out by recently developed amperometric bioelectrodes, on the interactions between carbonic anhydrase (CA) and the decarboxylating enzymes arginine decarboxylase (ADC), L-lysine decarboxylase (LDC), and L-ornithine decarboxylase (ODC). These are all pyridoxal phosphate dependent enzymes and catalyze the decarboxylation reaction of the respective amino acids, to give carbon dioxide and the corresponding diamine (agmatine, cadaverine, and putrescine, respectively). The rate of each decarboxylase catalyzed reaction was measured by monitoring the production of the respective diamine by a plant tissue diamino oxidase (DAO) based bioelectrode. DAO is the enzyme which catalyzes the oxidation of agmatine, cadaverine, and putrescine with the production of NH and H2O2. DAO-based bioelectrodes consist of an amperometric H2O2 electrode, coupled to the biocatalytic membrane formed by a whole plant tissue (lentil cotyledon) containing the enzyme DAO, immobilized on a dialysis membrane by polyazetidine prepolymer (PAP). The bioelectrodes were calibrated and characterized in standard solutions of agmatine, cadaverine, and putrescine. Kinetic studies to measure decarboxylase activity were performed in the presence of different concentrations of ADC, LDC, and ODC, resulting in a lowest detection limit of 10, 25, and 10 U l(-1), respectively. The effect of bovine CA II (bCAII) was evaluated in the presence of 500 U l(-1) of each decarboxylase, showing a marked increase of the rate of the decarboxylation reaction. These results suggest that (i) CA can be used to enhance the performance of decarboxylase-based biosensors, and (ii) it possibly plays further physiological roles, acting synergistically, at specific cellular and subcellular sites, with low-activity decarboxylating enzymes. PMID- 10379570 TI - Unmodified supported thiol/lipid bilayers: studies of structural disorder and conducting mechanism by cyclic voltammetry and AC impedance. AB - Supported thiol/lipid bilayer assembly, one of the most spectacular bilayer systems in recent years, has provided a good model to study biomembranes because of its high mechanical stability. In this work, the structural and conducting property of unmodified Au supported octadecanethiol/phosphatidylcholine bilayers were investigated using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The forming process of bilayer was monitored by capacitance plane plot. The normalized membrane capacitance of supported bilayer is 0.52 microF cm(-2). Kinetically controlled voltammograms determined by Butler-Volmer equation were obtained for both thiol monolayer and thiol/lipid bilayer in linear sweep voltammetry. Results of EIS experiment indicate that collapsed sites and pinhole defects exist in thiol monolayer and lipid monolayer, respectively. The difference between the values of experimental and theoretical standard electron transfer rate constant indicates that the conducting mechanism of Au supported thiol monolayer is electron tunneling at collapsed sites. The conducting mechanism of Au supported thiol/lipid bilayer is attributed as the following: the electroactive species could diffuse through pinholes in the lipid monolayer and reach collapsed sites in thiol monolayer, where electron transfer occurs via a tunneling process. The fractional coverage of the lipid monolayer measure by EIS experiments is about 0.98 or higher. PMID- 10379571 TI - Real-time monitoring of enzymatic cleavage of nucleic acids using a quartz crystal microbalance. AB - The use of quartz crystal microbalance (QCM) for monitoring in situ the enzymatic cleavage of surface-confined nucleic acids by nucleases is described. Such real time monitoring of mass changes associated with the enzymatic digestion indicates that the activity and specificity of nucleases is preserved at the gold surface, and can be used for manipulating surface-confined DNAs and RNAs. These observations indicate great promise for using QCM for elucidating the interactions of nucleic acids with enzymes, and for enhancing the power of hybridization biosensors. PMID- 10379572 TI - Low frequency alternating electric fields inhibit lactose uptake in Kluyveromyces marxianus. AB - Frequency-dependent lactose uptake via the H+/lactose symporter in an externally applied low-intensity alternating electric field was demonstrated, using tracer flux experiments. The uptake of radiolabeled lactose was significantly inhibited with the electric field-strength of 30 V/cm and at frequencies below 10 Hz. PMID- 10379573 TI - Examination of the relationship between parameters to determine electropermeability of Saccharomyces cerevisiae. AB - A rectangular electric pulse was applied to Saccharomyces cerevisiae suspensions in NaCl solutions. The relationship between field strength, pulse width and conductivity of extracellular media of key--factors to determine the yield of electropermeability--was examined at the time when the same permeability occurred. The results showed that the dependence of the yield of permeability upon the width of applied pulse was mutually related with the conductivity of extracellular media. Namely at one field strength, the value of pulse width is inversely proportional to that of conductivity of media and its relationship holds true for any field strength. Further, the relationship between parameters considered bears a close resemblance to that recognized between stress amplitude and the number of cycles to failure in the fatigue fracture of materials. PMID- 10379574 TI - In vivo recording from identifiable neurons of the locomotor network in the developing zebrafish. AB - The zebrafish is a popular model for developmental studies due to its accessibility by cellular, molecular and genetic approaches. As a complement to these other methods, we have devised an exposed hindbrain/spinal cord preparation in the curarized zebrafish embryo and larva that permits intracellular labeling and patch clamp recording from individually identified sensory neurons, motoneurons and interneurons in vivo. Regular bursts of synaptic potentials and action potentials were observed under whole-cell current clamp in embryonic motoneurons and in some identified interneurons. Larval neurons showed prolonged depolarizations with synaptically driven bursts of action potentials. Frequent spontaneous synaptic potentials were observed and synaptic currents were effectively space clamped. It is thus feasible to study in vivo the properties of identifiable neurons of the developing locomotor network in the zebrafish, including their synaptic activity, firing patterns and interconnections. PMID- 10379575 TI - Induction of hyperphosphorylated tau in living slices of rat hippocampal formation and subsequent detection using an ELISA. AB - Although hyperphosphorylated tau is an established feature of Alzheimer's Disease, its role in the disease process is poorly understood, partly because of lack of suitable animal models. We describe the use of living slices of rat hippocampal formation to study tau phosphorylation. Using the AT8 antibody in an ELISA, phosphorylated tau was detected in freshly frozen slices and it increased significantly in slices that were incubated in an electrophysiological recording chamber; the amount detected was greatest when the homogenisation buffer contained phosphatase and kinase inhibitors. The phosphorylated tau content of the slices increased significantly after exposure to the phosphatase 1 and 2A inhibitor okadaic acid (OA) - 1.5 microM. Electrophysiological recordings confirmed that slices were alive and that OA had no acute toxic effect. In control slices phosphorylated tau, detected immunohistochemically, was mainly in the somatodendritic compartment of neurones; in OA treated slices, there was an apparent decrease in somatodendritic AT8 staining and an increase in neuropil staining. Our system enables the induction of hyperphosphorylated tau within living slices, in an experimental environment that can be used to study the biological consequences of such a change, and may therefore help further our understanding of the significance of hyperphosphorylated tau in Alzheimer's Disease. PMID- 10379576 TI - Effect of temperature and calcium on transneuronal diffusion of DiI in fixed brain preparations. AB - The lipophilic tracer 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) was used to label neuronal pathways in fixed goldfish brains. The normal procedure involving 4% paraformaldehyde as a fixative, applying DiI and storing the brain in the fixative at 40 degrees C resulted in many cases in a rather diffuse labeling of fiber pathways and the occurrence of transneuronally labeled cells and fibers. We found that calcium and heat both facilitate the diffusion of DiI out of membranes in vibratome sections. We modified the protocol by adding the calcium binding substance ethylenediamine tetraacetate to all solutions and incubated at room temperature. This improved the sharpness of labeled structures and eliminated the transneuronal labeling in our material. Although transneuronal transport of the tracer may still occur under certain circumstances, the present modification of the DiI staining procedure substantially increased the staining quality and reproducibility and decreased the occurrence of transneuronal labeling. PMID- 10379577 TI - A multichannel system for recording and analysis of cortical field potentials in freely moving rats. AB - A system has been developed to record and analyze the cortical electrical activity from 16 different sites in freely moving rats. The hardware includes a 16-channel amplifier system whose high input impedance, low noise, small size, light weight and shielded multistrand connecting cable allow high quality multichannel recording of field potentials. The software developed for this system consists of data acquisition, data analysis and topographic mapping of cortical-evoked potentials as well as electroencephalograms. Cortical field potentials evoked by CO2-laser stimulation were compared between wakeful and pentobarbital-treated conditions. To investigate the background interference produced by sleep spindle, three kinds of reference-free methods (the Wilson, local average and weighted average methods) were utilized to compare the coherence between field potentials obtained from two cerebral hemispheres using monopolar vs. reference-free recordings. PMID- 10379578 TI - Recombinant adenovirus is an appropriate vector for endocytotic protein trafficking studies in cultured neurons. AB - Endocytosis of full-length beta-amyloid precursor protein (APP) from the plasma membrane contributes to beta-amyloid peptide (Abeta) secretion, and, hence, potentially contributes to the molecular pathogenesis of Alzheimer's disease. We recently have demonstrated that central neuronal APP is endocytosed in a common vesicular compartment with recycling synaptic vesicle integral membrane proteins, but is then sorted away from synaptic vesicles for retrograde transport to the neuronal soma. For this report, we explore whether recombinant adenovirus can be used to modulate APP expression in cultured central neurons to study APP processing by the endocytotic pathway in these cells. Using a replication deficient recombinant adenovirus that expresses a lacZ reporter (Ad5/CMV-lacZ), we demonstrate high efficiency of transfection (30-35%) at low viral titer (10-20 MOI), with no significant neuronal toxicity or cytoarchitectural change. In addition, we demonstrate that infection with the control virus does not result in re-direction of endogenous neuronal APP from usual endocytotic pathways. We have prepared, using the same genomic background as the control virus, an adenoviral vector that expresses the neuronal isoform of human APP (Ad5/CMV-APP). Infection with Ad5/CMV-APP at 10-20 MOI results in significantly increased immunoreactivity for endocytosed APP with preservation of usual endocytotic trafficking. These results demonstrate that recombinant adenovirus at low titer is an appropriate and effective vector for protein trafficking/processing studies in cultured central neurons. PMID- 10379579 TI - Tyramide signal amplification in brain immunocytochemistry: adaptation to electron microscopy. AB - The tyramide signal amplification (TSA) technique is well-established in light microscopic immunohistochemistry and in situ hybridization to improve the signal to-noise ratio. The present study deals with its adaptation to the electron microscopic level using the pre-embedding technique and a modified protocol. The outcome of immunolabeling of most of the antigens tested in brain tissue, including endothelial and neuronal nitric oxide synthase, glial fibrillary acidic protein, and isolectin B4, was greatly improved. If signal amplification is required, the TSA-technique proved to be reliable with high specificity and good ultrastructural resolution. PMID- 10379580 TI - Quantitation of multiple gene expression by in situ hybridization autoradiography: accurate normalization using Bayes classifier. AB - In the method of in situ hybridization autoradiography, quantitative comparisons among multiple mRNA signals have proven difficult for many reasons, attributable both to technical factors (e.g. different probe specific activities) as well as to large differences in the patterns and levels of expression of different genes in pathologic states. Here we report a standardized normalization procedure for in situ hybridization autoradiography, employing a Bayes classifier, which permits the comparison of multiple mRNA probes. Autoradiograms of different probes in individual animals are first digitized and converted to units of radioactivity. Next, pixel-distribution histograms are generated for each mRNA signal. The Bayes classifier is then used to establish an optimal threshold to distinguish activated and non-activated pixels. This threshold also defines the minimal level of mRNA expression. The maximal mRNA signal is defined as the mean + 3 SD of the activated pixel distribution. We then use a linear transformation to convert each pixel from absolute activity to percentage of maximal mRNA signal for that particular probe. The normalized autoradiographic images can then be averaged to represent group trends and can be compared by standard statistical methods. We illustrate this normalization procedure using in situ hybridization autoradiography for three genes (GADD45, HSP70 and MAP2) expressed in the brains of rats studied at various recirculation times following transient (2 h) middle cerebral artery occlusion. The Bayes classifier is reviewed and its analytical application is presented. Step-by-step examples of intermediate steps are presented, construction of averaged data sets, and pixel-based statistical comparisons among expressed genes. PMID- 10379581 TI - No difference between estimated mean nuclear volumes of various types of neurons in the mouse brain obtained on either isotropic uniform random sections or conventional frontal or sagittal sections. AB - Whenever using modern stereological methods for estimating number-weighted or volume-weighted mean volumes of biological particles such as cell nuclei, either 'isotropic uniform random' (IUR) tissue sections or 'vertical' ones had to be used. However, with the currently available procedures and tools it was virtually impossible to prepare such sections from small specimens such as the mouse brain. Here, a modification of the 'isector' is presented, which allows the embedding of mouse brain halves into paraffin spheres as a useful basis for preparing IUR sections. By using this modified isector it could be shown for various types of neurons in the hippocampus and cerebellum of young adult mice, that there are no differences between estimated mean nuclear volumes obtained on IUR sections and those obtained on conventional frontal or sagittal ones. This result may be used to expand the interpretation of estimated mean nuclear volumes of the types of neurons investigated here. PMID- 10379582 TI - Rapid fabrication of plastic-insulated carbon-fiber electrodes for micro amperometry. AB - Carbon-fiber amperometry and voltammetry are useful techniques to measure secretion of oxidizable neurotransmitters from neurosecretory cells. Recent applications with probes of small geometry permit detection of the exocytosis of single secretory vesicles in individual cells. We have developed a semi-automatic puller and cutter to prepare such plastic-insulated electrodes efficiently with various sizes of carbon fibers. The electrodes are smooth, reproducible, and easy to make. PMID- 10379583 TI - An improved heterodyne laser interferometer for use in studies of cochlear mechanics. AB - A displacement-sensitive heterodyne laser interferometer is described which is suitable to measure the sound-evoked motion of various structures in the intact, living cochlea. Data are included to demonstrate the interferometer's low noise floor ( < 1 pm Hz (-0.5)), wide bandwidth (from d.c. to 63 kHz) and linear dynamic range ( > 300 microm). The interferometer's optical sensitivity is sufficient to detect the motion of the basilar membrane and various components of the organ of Corti without the use of artificial reflectors: the reflectivity needed to achieve a 10 pm Hz(-0.5) noise-floor is approximately 0.0001%, and the optical sectioning depth is approximately 23 microm. These features make the interferometer ideally suited to measure acoustic vibrations in the living inner ear, but there may also be applications in other fields of neuroscience. PMID- 10379584 TI - Characterization of a recovery global cerebral ischemia model in the mouse. AB - Transgenic/knockout murine variants allow roles of specific proteins to be studied in cerebral ischemia. Because of the size of mice, however, study of prolonged recovery from global ischemia has been limited. This project characterized an adaptation of the rat two-vessel occlusion model of global ischemia for use in the mouse. C57B1/6J mice (8 weeks old; 21 +/- 1 g) were overnight fasted, anesthetized with halothane, intubated and mechanically ventilated. The right internal jugular vein and femoral artery were cannulated. Pericranial temperature was held at 37.0 degrees C. The carotid arteries were occluded and mean arterial pressure was reduced to 35 mmHg with 0.3 mg intra arterial trimethaphan and venous exsanguination. Electroencephalographic isoelectricity was confirmed in cohort mice. Ten minutes later ischemia was reversed. Mice were allowed 1, 3 or 5 days survival followed by histologic analysis. Regional cerebral blood flow (CBF) was determined autoradiographically. Outcome effects of intra-ischemic hyperglycemia (approximately 350 mg/dl) or hypothermia (34 degrees C) were also examined. The mortality rate was less than 10% in all recovery groups. Ischemia caused reduction of CBF to < 2% of sham values in cortex, hippocampus, and caudoputamen. CBF was unchanged in thalamus, brainstem and cerebellum. CA1 damage, greater after 3 days vs. 1 day reperfusion, was not further increased at 5 days. Histologic injury was increased by hyperglycemia although seizures did not occur. Hypothermia reduced CA1 damage. This study demonstrates feasibility of using the two-vessel occlusion + hypotension recovery model in the mouse. Recovery intervals of > or = 3 days are required to account for delayed CA1 neuronal necrosis. Histologic outcome can be modulated by known physiologic determinants of ischemic brain damage. PMID- 10379585 TI - Postoperative aneurysm remnants: endovascular treatment as an alternative to further surgery. AB - Because further surgery on postoperative aneurysm remnants can be difficult and lead to significant morbidity and mortality, endovascular treatment, using controlled detachable coils, was performed in three patients with such remnants. The endovascular approach was technically more difficult in these cases than in previously untreated patients. In one case, the "remodelling" technique was necessary. Given the successful outcome in these patients, endovascular treatment can be proposed as an alternative to another operation, when further surgery appears too risky or is refused by the patient. PMID- 10379586 TI - Phenytoin as a liquid material for embolisation of tumours. AB - We carried out embolisation with phenytoin in seven patients with meningiomas; in three by cannulation of the middle meningeal artery during surgery and in four by microcatheter introduced into the middle meningeal artery. Phenytoin (125-500 mg) diluted with saline (25 mg/ml) was injected slowly as a bolus. There was no evidence of devascularisation in two meningiomas treated with 125 mg and 150 mg of phenytoin. Large areas of ischaemic and haemorrhagic necrosis were found in five meningiomas treated with 250-500 mg. After embolisation, the tumour blush disappeared, with preservation of the feeding arteries. Total resection of the tumour was performed with minimum blood loss. Caution and proper use are emphasised to avoid increase in tumour volume and reflux. Phenytoin could be a suitable material for superselective embolisation. PMID- 10379587 TI - Post-procedure migration of Guglielmi detachable coils and Mechanical detachable spirals. AB - We describe a previously unreported complication of the use of Guglielmi detachable coils and Mechanical detachable spirals in endovascular treatment of intracranial aneurysms. We document four cases in which migration of part of a coil into the parent artery occurred after completion of the procedure. Possible mechanisms are discussed. PMID- 10379588 TI - Transvenous Doppler guidewire sonographic monitoring during treatment of a complex vertebral arteriovenous fistula associated with neurofibromatosis type 1. AB - A Doppler sonographic guidewire was used to monitor incremental changes in draining vein (DV) flow during endovascular occlusion of a complex vertebral arteriovenous fistula (AVF) in a patient with neurofibromatosis type 1. Transvenous monitoring of average peak velocity (APV) and the maximum-minus minimum peak velocity (MxPV-MnPV) demonstrated a progression from a highly pulsatile, fast flow before embolization to a nonpulsatile, slow flow indicating a successful occlusion of the AVF (hemodynamic endpoint of treatment). Prior to this, apparent angiographic occlusion of the AVF was thought to signify a successful endpoint; however, persistently elevated values for APV and MxPV-MnPV in the DV signalled the presence of an additional contralateral arterial contribution. Transvenous monitoring of flow velocity appears to be ideally suited to establishing a hemodynamic endpoint of embolotherapy in the presence of complex arteriovenous shunting, as may occur with the vasculopathy of neurofibromatosis. PMID- 10379589 TI - Presumed intraventricular meningioma treated by embolisation and the gamma knife. AB - A 58-year-old woman with a presumed incidentally discovered meningioma in the left lateral ventricle was treated by superselective embolisation and gamma knife therapy. The diameter of the tumour was 40 mm, and its main feeding artery was the left lateral posterior choroidal artery. This vessel was embolised with microcoils. At 8 months following embolisation, the diameter of the tumour had decreased and was stable. The gamma knife was chosen as an adjuvant therapy for the further control 13 months after embolisation. Embolisation and gamma knife therapy may be an alternative treatment for meningiomas where surgical resection appears difficult. PMID- 10379590 TI - Experimental side-wall aneurysms: a natural history study. AB - We studied the natural history of canine side-wall experimental aneurysms to determine the incidence of spontaneous aneurysm thrombosis, to serve as control data for future studies focusing on development of aneurysm occlusion devices. Bilateral common carotid artery vein patch aneurysms were surgically created in eight mongrel dogs (20-25 kg). Duplex Doppler sonography was performed at 14 days and angiography between 30 and 210 days following aneurysm creation. Sonography demonstrated patency of 13 (81%) of 16 aneurysms. Patent aneurysms ranged in size from 8 x 10 mm to 14 x 16 mm. Conventional angiography was performed in four dogs approximately 30 days following aneurysm creation; in these four, all of 7 initially patent on sonography remained fully patent. One dog underwent conventional angiography at approximately 60 days following aneurysm creation; both aneurysms in this case remained widely patent. Three dogs underwent conventional angiography at approximately 200 days following aneurysm creation; all 4 aneurysms initially patent on sonography remained fully patent. None of the three aneurysms found to be occluded on sonographs demonstrated spontaneous recanalization. The canine side-wall aneurysm model is a valid tool for testing some aneurysm-occlusion devices, because control aneurysms remain patent indefinitely. PMID- 10379591 TI - Fast FLAIR sequence for detecting major vascular abnormalities during the hyperacute phase of stroke: a comparison with MR angiography. AB - In the hyperacute phase of stroke, occluded vessels can be seen as high signal on fast-FLAIR images or as absence of flow-related enhancement in maximum-intensity projection (MIP) MR angiography (MRA). To compare these techniques, we examined 53 patients within 6 h of a stroke, using a standardised MRI protocol including fast-FLAIR and 3D time-of-flight TOF MR to detect vessel occlusion or reduced flow corresponding to the suspected ischaemic territory. Brain infarcts were confirmed on MRI after 1-5 days in 41 cases (77%). The overall accuracy of 3D-TOF MRA was 68% and sensitivity, specificity, positive and negative predictive values were 67%, 71%, 87%, and 43% respectively. Values for the fast-FLAIR sequence were: 65%, 85%, 93% and 44%, with an overall accuracy of 70%. The fast-FLAIR sequence was thus able to show occluded vessels or reduced flow with about the same accuracy as 3D-TOF MRA and enabled better prediction of the ischaemic area. PMID- 10379592 TI - Interobserver agreement for diagnostic MRI criteria in suspected multiple sclerosis. AB - MRI is the paraclinical test most widely used to support the diagnosis of multiple sclerosis (MS). We evaluated interobserver agreement in applying diagnostic criteria to MRI obtained at first presentation. Five experienced observers scored 25 sets of images consisting of unenhanced T2- and gadolinium enhanced T1-weighted images (approximately half the sets were normal). We scored frontal, parietal, temporal, occipital, infratentorial and basal ganglia lesions and the total number of lesions on T2-weighted images; periventricular, callosal, juxtacortical and ovoid lesions and those > 5 mm in maximum diameter; contrast enhancing and hypointense lesions. Based on a combination of imaging findings patients were classified as compatible or not compatible with MS according to composite criteria. Observer concordance was characterised by weighted kappa values (kappa) and mean average difference to the median (MADM) scores. Using the raw scores, there was poor agreement for the total number of lesions on T2 weighted images, and for occipital, oval, juxtacortical and hypointense lesions. Moderate agreement was found for frontal, callosal, basal ganglia and large lesions on T2 weighting. Good agreement was attained for parietal, temporal, infratentorial and periventricular lesions. After dichotomisation according to accepted cut-off values, most criteria performed better, especially the number of lesions on T2-weighted images (P < 0.05). Good agreement was found for the criteria of Paty and Fazekas and moderate agreement for those of Barkhof. While experienced observers may not agree on the total number of lesions, they show quite good agreement for commonly used cut-off points and elements in the composite criteria. This validates the use of MRI in the diagnosis of MS, and the use of dichotomised and composite criteria. PMID- 10379594 TI - Apraxia of speech associated with an infarct in the precentral gyrus of the insula. AB - It has been postulated that the precentral gyrus in the left insula is responsible for co-ordination of speech. We report a patient with this disturbance who showed an acute infarct limited to this region. PMID- 10379593 TI - Familial tumoral calcinosis: association with cerebral and peripheral aneurysm formation. AB - Two siblings with histologically and radiologically proven tumoral calcinosis presented with cerebral and peripheral aneurysms. The brother died of a ruptured subclavian artery aneurysm after surgical repair of brachial, iliofemoral and coeliac axis aneurysms. Magnetic resonance and catheter angiography in the sister demonstrated marked carotid dysplasia and a left ophthalmic segment aneurysm, not amenable to treatment. We believe this is the first reported case of familial aneurysms in association with tumoral calcinosis. PMID- 10379595 TI - MRI of pituitary adenoma with extensive amyloid formation. AB - We report a patient with a pituitary adenoma with extensive amyloid formation. T2 weighted MRI was most characteristic for amyloid deposition. PMID- 10379596 TI - Spinal disease in neurologically symptomatic HIV-positive patients. AB - We review the MRI findings of human immunodeficiency virus (HIV)-positive patients with "spinal" symptoms and review the literature. In 23 consecutive HIV positive patients presenting with acute neurologic complaints thought to be referable to the spine, we reviewed spinal MRI, medical charts, and laboratory, pathologic, and autopsy data. In the early stages of HIV infection, the common causes of spinal complaints (i. e., degenerative spine and disc disease) predominated. However, pathology may be missed without contrast-enhanced MRI of the spine. In more advanced cases, the differential diagnosis includes one or more neoplastic and/or infectious causes which require contrast-enhanced MRI for detection. In these cases, normal cerebrospinal fluid findings should not preclude contrast-enhanced MRI of the spine. Imaging of the brain may also be indicated in cases when the spinal study is negative. PMID- 10379597 TI - Intracranial contrast-enhancing masses in infants with capillary haemangioma of the head and neck: intracranial capillary haemangioma? AB - Contrast-enhancing intracranial masses are rarely found in infants with extracranial capillary haemangiomas (CH). We aimed to assess their nature and progression in three patients undergoing CT and/or MRI. The changes in size of both extra- and intracranial lesions were recorded. In a fourth case, a single examination was obtained. All patients harboured one or two enhancing intracranial nodular, meningeal-based lesions. Diffuse leptomeningeal enhancement of the cerebellar surface was also seen in one, which disappeared at follow-up. In all but one of the cases, the intracranial lesions were on the same side as the extracranial CH. These lesions and the extracranial CH demonstrated parallel changes in size (suggesting that both represent CH) during follow-up of 1-2 years: the size of intracranial lesions and the extracranial CH decreased in two cases, whereas it was unchanged in the third. One patient had a persistent trigeminal artery, while another had cerebellar atrophy with high signal in the cortex on T2-weighted images. In some cases, extracranial CH are part of PHACE syndrome; the association with intracranial CH might represent a peculiar phenotype of this rare vascular phakomatosis. As extracranial CH are known to regress spontaneously in the majority of cases, a conservative approach is recommended also for presumed intracranial CH; surgery should be avoided unless follow-up studies demonstrate growth. PMID- 10379598 TI - Infantile neuroaxonal dystrophy: neuroradiological studies in 11 patients. AB - We report the imaging findings in 11 patients with infantile neuroaxonal dystrophy. Ten patients underwent 15 MRI examinations; one patient had only CT. Of the ten patients who underwent MRI, eight had cerebellar atrophy and mildly increased signal from the cerebellar cortex on T2-weighted images. With T2 weighting there was slightly increased signal from the dentate nuclei in two patients and from the posterior periventricular white matter in three. We saw four patients with a thin optic chiasm. The only two brothers in the series had markedly low signal from the globus pallidus and substantia nigra on 1.5 T T2 weighted images, as seen in Hallervorden-Spatz disease (HSD). Abnormalities of the globus pallidus may be related to a protracted course of the disease. However, an overlap with HSD should be considered. PMID- 10379599 TI - "Growing fontanelle": a serious complication of difficult vacuum extraction. AB - Growing skull fractures in combination with leptomeningeal cysts are well known in childhood. A rare case of a growing fontanelle due to a leptomeningeal cyst is presented. The cyst occurred due to a traumatic delivery with vacuum extraction. Operative repair of the cyst revealed a dural tear at the border of the fontanelle. The imaging findings are discussed. PMID- 10379600 TI - Imaging of the normal pontine cisternal segment of the abducens nerve, using three-dimensional constructive interference in the steady state MRI. AB - Our objective was to determine the visibility of the cisternal segment of the normal abducens nerve using a three-dimensional Fourier-Transform constructive interference in the steady state (3DFT-CISS) sequence. Its visibility was rated in 150 patients without clinical evidence of abducens nerve disturbance. Axial 1 mm 3DFT-CISS images were obtained (TR/TE 17/7 ms, flip angle 50 degrees, field of view 160 mm, matrix 256 x 256). The cisternal segment was seen in 79% of cases, bilaterally in 73% and unilaterally in 11% of cases; neither cisternal segment was seen in 16% of cases. Identification of Dorello's canal was often of help in detecting the point lateral to the dorsum sellae at which the nerve pierces the dura mater. Flow artifacts and vascular loops in the pontine cistern sometimes caused problems in interpretation. 3DFT-CISS MRI with 1-mm-thick sections can however be considered a reasonably reliable technique for showing the cisternal segment of the abducens nerve. PMID- 10379601 TI - Subacute combined degeneration of the spinal cord: demonstration of contrast enhancement. PMID- 10379602 TI - 1998 Jules Gonin lecture of the Retina Research Foundation. Drug treatment of ocular neovascularization and proliferation. PMID- 10379604 TI - Optic disc changes in normotensive persons with unilateral exfoliation syndrome: a 3-year follow-up study. AB - BACKGROUND: If, at the time of glaucoma diagnosis, the intraocular pressure (IOP) is higher and the initial field loss more advanced in glaucomatous eyes with than without exfoliation, the cause of the optic disc damage has been suggested to be the high IOP associated with the exfoliation syndrome (EXS). We decided to investigate whether EXS alone, without the contributory effect of measured raised IOP, is a risk factor for optic nerve damage. METHODS: Twenty-two non glaucomatous, normotensive persons with clinically unilateral EXS were examined for IOP, visual fields (Octopus G1) and disc topography (Imagenet, Topcon) and followed up for 3 years. RESULTS: At the start, the paired exfoliative (E) and non-exfoliative (NE) eyes did not differ in IOP, disc, rim, or cup areas, or cup volumes. They differed in R/D (rim/disc) radius ratio in the inferior section of the optic disc. During the follow-up period, the IOP increased in the E and the NE eyes, and changes indicative of nerve fiber loss were measured in both eyes. In those (n=14) in whom the IOP in the two eyes was equal throughout the follow up period, disc changes took place only in the E eye. CONCLUSION: The exfoliative process in itself may be a risk factor for optic disc changes. PMID- 10379603 TI - Evaluation of microvascularization pattern visibility in human choroidal melanomas: comparison of confocal fluorescein with indocyanine green angiography. AB - BACKGROUND: The presence of specific microvascularization patterns (networks, parallel with and without crosslinking, silent) in histological sections of human choroidal melanomas has prognostic significance for survival. We showed previously in selected patients that the identification of these microvascularization patterns is possible in vivo by using confocal scanning laser indocyanine green angiography and that this technique is superior to fluorescein angiography using a conventional acquisition technique with a fundus camera. We now routinely use simultaneous confocal fluorescein/indocyanine green angiography to study microvascularization patterns in choroidal melanomas. The purpose of this study was to compare the visibility of tumor vessels and microvascularization patterns in fluorescein and indocyanine green angiography in simultaneous confocal series taken with the same instrument in a large prospective series of patients. PATIENTS AND METHODS: The simultaneously procured confocal fluorescein and indocyanine green angiograms of 50 patients with untreated choroidal melanomas (maximal apical height according to standardized A scan between 2 and 8 mm) were studied for the visibility of tumor vessels and microvascularization patterns. At least one simultaneous confocal optical series (32 images in sequential depth order) during the early arterial venous phase was obtained per patient. RESULTS: Confocal forescein angiography disclosed signs of tumor vascularization in 12 (24%) of the 50 patients examined. However, in only 3 patients (6%) could microvascularization patterns be identified using confocal fluorescein angiography, and only in the very early arterial phase, which is often difficult to capture. In contrast, simultaneously obtained confocal indocyanine green angiograms disclosed tumor vessels in 47 (94%) of the examined 50 patients and microvascularization patterns could be identified in all of these cases. In 3 patients (6%) no tumor vessels could be detected within the tumor borders. CONCLUSION: This study demonstrates that confocal indocyanine green angiography images microvascularization patterns in choroidal melanomas better than fluorescein angiography, even when the images are acquired with the same technique. This can be explained with the different absorption, fluorescence and exudation characteristics of these dyes. In vivo imaging of these microvascularization patterns using confocal indocyanine green angiography offers the possibility of assessing the prognosis of choroidal melanomas without the removal of tissue. PMID- 10379605 TI - Risk factors for the progression of treated primary open-angle glaucoma: a multivariate life-table analysis. AB - BACKGROUND: Several factors have been reported as risk factors for the progression of primary open-angle glaucoma (POAG) but previous reports were not necessarily in agreement. We applied a multivariate life-table analysis to a large number of longitudinal data to determine the extent of the influence of various factors simultaneously. METHODS: Two hundred fifteen eyes of 215 POAG patients were included. The follow-up period ranged from 24 to 134 months (average 82.7 months). The visual field stage was determined separately in upper and lower hemifields according to the classification of Aulhorn (modified by Greve). The progression was defined as an irreversible increase of the stage in at least one hemifield. The follow-up data were analyzed with the Cox proportional hazard model. RESULTS: Mean intraocular pressure (IOP) in the follow up period and the initial visual field stage significantly affected POAG progression (P<0.05). The risk of POAG progression was calculated to double as the mean IOP increased by 4 mmHg. Eyes with the initial visual field of stage 0-1 and moderately advanced stages had a greater risk of progression than other stages. CONCLUSION: To prevent the progression of POAG, the IOP should be kept as low as possible, particularly at the early and moderately advanced stages. PMID- 10379607 TI - Retinal nerve fiber layer thickness in human eyes. AB - BACKGROUND: A study was carried out to measure the thickness of the retinal nerve fiber layer (RNFL) at the optic disc border. METHODS: RNFL thickness at the optic disc border was histomorphometrically measured on histological sections of 22 human eyes with normal optic nerves and 21 human eyes with absolute secondary angle-closure glaucoma. For three eyes with normal optic nerves, serial sections through the whole optic disc area were available. RESULTS: In the eyes with normal optic nerves, the RNFL at the optic disc border showed a double hump configuration with the highest mean thickness in the inferior quadrant (mean +/- S.D: 266+/-64 microm), followed by the superior quadrant (240+/-57 microm), the nasal quadrant (220+/-70 microm), and finally the temporal quadrant (170+/-58 microm). In the three globes with serial sections, RNFL was thickest at the inferior disc pole (397+/-58 microm), followed by the superior disc pole (313+/ 38 microm), the nasal disc pole (165+/-19 microm), and finally the temporal disc pole (131+/-15 microm). In the eyes with absolute glaucoma, mean thickness of the remainder of the RNFL was 40+/-18 microm with no marked differences between the disc regions. CONCLUSIONS: In normal eyes, the RNFL shows a double hump configuration with its thinnest part at the temporal disc pole, followed by the nasal disc pole and the superior disc pole. RNFL is thickest at the inferior disc pole. In glaucomatous optic neuropathy, the inner limiting membrane moves backward about 60-100 microm at the temporal disc border, and more than 200 microm at the inferior and superior disc poles. PMID- 10379606 TI - Pterygium with bulbar conjunctival hemorrhages. AB - BACKGROUND: This study was carried out to describe the clinical features and electron-microscopic characteristics of the capillaries in pterygium with conjunctival hemorrhage. METHODS: We compared the clinical findings in Japanese and Tunisian primary pterygia and in pterygium with and without bulbar conjunctival hemorrhages. The capillary fine structures of pterygium with bulbar conjunctival hemorrhages were studied by electron microscopy and compared with those without bulbar conjunctival hemorrhages. RESULTS: Conjunctival hemorrhages were noted in 16.4% of Tunisian patients but not in any Japanese patients. The former pterygia were significantly larger, more vascular and more frequently hyperemic than the latter. Pterygia with bulbar conjunctival hemorrhages were more hyperemic than without hemorrhages. Electron microscopy revealed interruptions of the endothelial cells and basement membrane of the capillaries in the pterygia with bulbar conjunctival hemorrhages, with blood cells escaping through the endothelial interruptions. CONCLUSION: Conjunctival hemorrhage in pterygia may be caused by fragility of the endothelial cells and basement membrane in the capillaries, which easily induces hemorrhage when the eyes are irritated by rubbing or by conjunctival foreign bodies. Pterygium with conjunctival hemorrhage is more frequently found in Tunisians than in Japanese. PMID- 10379608 TI - Blood coagulation parameters in retinal arterial occlusion. AB - BACKGROUND: Thromboembolism is considered a crucial event in the pathogenesis of retinal occlusion, resulting in a severe damage of central or peripheral visual function. METHODS: We evaluated hemostatic system parameters in the plasma of 14 patients (11 males and 3 females aged 59-73 years) affected by acute retinal ischemia (central retinal arterial occlusion or arterial branch occlusion). The diagnosis of retinal arterial occlusion was established according to clinical symptoms, ophthalmoscopic findings and fluorescein angiography. In addition to routine coagulation tests, antithrombin III, prothrombin fragment 1+2 (F1+2), thrombin-antithrombin III complex (TAT), and D-dimer were measured in the plasma both at the moment of diagnosis (before therapy initiation) and 3-6 months later (at least 1 months after antithrombotic therapy discontinuation). RESULTS: We found a marked increase in the plasma levels of F1+2, TAT, and D-dimer during the acute event, compared with healthy control values. F1+2 and TAT significantly decreased during follow-up, whereas D-dimer was unchanged. CONCLUSION: Our data indicate accelerated conversion of prothrombin to thrombin (high F1+2) and an increase in circulating thrombin (high TAT) during retinal arterial occlusion, providing evidence that increased thrombin generation may play a role in acute retinal ischemia. PMID- 10379609 TI - Regional distribution of optic nerve head blood flow. AB - BACKGROUND: Advanced glaucoma typically results in damage of the temporal neuroretinal rim. As vascular factors are of pathogenic importance in the development of glaucomatous damage, the present study investigated whether regional differences in perfusion might be the reason for the preferential damage of the temporal neuroretinal rim. MATERIAL AND METHODS: Blood flow of the neuroretinal rim was measured with the laser Doppler flowmeter (LDF) Oculix 4000 (continuous measurement of an area of 160 microm diameter) and the Heidelberg retina flowmeter (HRF). Both instruments measure the capillary blood flow (flow), the relative velocity of erythrocytes (velocity) and the relative volume of moving erythrocytes (volume). We examined one randomly chosen eye of 55 healthy subjects without history of glaucoma aged 22-57 years (mean 30 years). Each subject was measured with the LDF and HRF, each time nasally and temporally, away from visible vessels. The intraocular pressure (IOP) was measured with the Goldmann tonometer. Heart rate and systolic and diastolic blood pressure were measured. RESULTS: The LDF measurements of the optic nerve head showed nasal flow of 12.4+/-5.6 AU and temporal flow of 9.8+/-3.6 AU. The HRF showed a nasal flow of 477+/-161 AU and a temporal flow of 368+/-166 AU. The volume measurements done by LDF showed nasally a value of 0.68+/-0.40 AU and temporally a value of 0.46+/ 0.21 AU. The HRF volume measurements showed nasal values of 16.1+/-4.3 AU and temporal values of 13.0+/-4.0 AU. The LDF velocity values were nasally 0.22+/ 0.05 kHz and temporally 0.26+/-0.05 kHz. HRF measurements showed velocity values of 1.7+/-0.5 kHz nasally and 1.3+/-0.6 kHz temporally. The differences were highly statistically significant for flow (LDF P=0.00007, HRF P=0.0005), volume (LDF P=0.00002, HRF P=0.00004) and velocity (LDF P=0.0002, HRF P=0.00004). The IOP was 12.6 mmHg. Blood pressure was 118/75 mmHg and the heart rate was 73 beats per minute. There was no correlation between age, IOP, BP and HR and the HRF/LDF measurements. CONCLUSION: The measurements with two different methodologies showed a decreased blood flow of the temporal neuroretinal rim compared to the nasal side. These local differences might be one reason for the preferential damage of the temporal neuroretinal rim in advanced glaucoma. PMID- 10379610 TI - Independent diagnostic value of fluorescein angiography in the evaluation of intraocular tumors. AB - BACKGROUND: Fluorescein angiography has been used in the clinical evaluation of suspected neoplastic lesions of the ocular fundus for over 30 years. Yet, the independent diagnostic value of this photographic technique in patients with suspected intraocular neoplasms has never been determined. METHODS: The authors evaluated color fundus photographs and fluorescein angiograms of 50 mass lesions of the ocular fundus. The cases were chosen to reflect a broad spectrum of lesions, including choroidal malignant melanoma, choroidal nevus, circumscribed choroidal hemangioma, metastatic carcinoma to the choroid, miscellaneous other neoplasms, hamartomas, or choristomas, and non-neoplastic lesions simulating neoplasms. Ten experienced retinal specialists independently reviewed the angiograms (presented in random order without the corresponding color fundus photographs) and rendered a diagnosis. Approximately 1 month later, each retinal specialist independently reviewed the color fundus slides (presented in a different random order without the corresponding fluorescein angiograms) and again rendered a diagnosis for each lesion. RESULTS: The accuracy of angiographic diagnosis by the different reviewers ranged from 16% to 56% (average 45.4%), while that based on review of the color fundus slides ranged from 32% to 78% (average 59.0%). This difference is statistically significant. CONCLUSION: Fluorescein angiography alone did not appear to be a reliable method for establishing the clinical diagnosis of neoplasms and simulating lesions of the ocular fundus. PMID- 10379611 TI - Effect of topical dorzolamide on optic nerve head blood flow. AB - PURPOSE: The topical carbonic anhydrase inhibitor dorzolamide has proven effective in lowering intraocular pressure in glaucoma patients. Because an impaired blood supply of the optic nerve has to be regarded as a major pathogenic risk factor it seems important to examine the effect of this new antiglaucomatous drug on capillary optic nerve head blood flow. METHODS: In a double-masked, randomized clinical trial, dorzolamide eye drops were applied to both eyes of 15 healthy subjects (8 female, 7 male, mean age 30.6 years) three times daily for 3 days. The control group (15 healthy volunteers, 9 female, 6 male, mean age 30.8 years) received a placebo preparation according to the same protocol. Intraocular pressure (IOP), blood pressure, heart rate, capillary optic nerve head blood flow and retinal blood flow were measured at baseline (1D0), 90 min after single instillation (1D90), and after 3 days of therapy (3D). Scanning laser Doppler flowmetry (Heidelberg Retina Flowmeter) and laser Doppler flowmetry according to Riva (Oculix 4000) were used to measure optic nerve head blood flow. RESULTS: IOP dropped in dorzolamide-treated subjects from 12.5 mmHg to 11.0/10.5 mmHg (1D0, 1D90, 3DO) and in the control group from 13.0 mmHg to 12.5/12.5 mmHg. Optic nerve blood flow as measured by scanning laser Doppler flowmetry showed no significant changes in dorzolamide-treated volunteers (temporal 310/329/315 AU, nasal 387/402/399 AU) or in the placebo group (temporal 238/306/276 AU, nasal 356/382/379 AU). Also as measured by laser Doppler flowmetry optic nerve head blood flow did not show significant changes in dorzolamide-treated volunteers (temporal 12.98/12.6/11.7 AU, nasal 16.6/16.9/15.7 AU) or in the placebo group (temporal 11.9/12.4/12.4 AU, nasal 16.1/15.8/17.7 AU). The systemic parameters blood pressure and heart rate remained unchanged during the treatment period. CONCLUSION: The results showed the expected drop in IOP. However, capillary optic nerve head blood flow, measured by two different techniques, did not change during therapy. This may be due to the effective autoregulation in human optic nerve head circulation, which seems not to be affected by dorzolamide. PMID- 10379612 TI - Anterior segment changes in rabbits after experimental aqueous replacement with various amounts of different perfluorocarbon liquids. AB - BACKGROUND: We evaluated biomicroscopic and histological effects on the anterior segment in the rabbit eye after temporary aqueous substitution with various amounts (0.2 cc and 0.025 cc) of perfluorodecaline (PFD) and perfluorophenanthrene (PFP). METHODS: A quantity of 0.2 cc of the two perfluorocarbon (PFC) liquids was exchanged simultaneously with about 50% of the aqueous in 15 rabbit eyes each for periods of 1, 2, or 4 weeks. At these points some eyes were enucleated for histological examination. After 2 and 4 weeks the substances were removed from the remaining eyes, which were then followed up for 8-10 weeks. In an additional 8 eyes, 0.025 cc PFD or PFP was injected and left for 8 weeks. Four eyes received balanced salt solution and served as controls. Beside biomicroscopic evaluation and measurement of the intraocular pressure, endothelial cell counts and corneal pachymetry were performed regularly during follow-up. RESULTS: The postoperative results were well comparable for PFD and PFP eyes. Within the first 2 weeks postoperatively corneal edema with endothelial cell loss was observed in both groups. Thereafter regression of the edema started independently of whether the substances were removed or not. IOP was not elevated at any time. At the end of follow-up central corneal thickness was the same as initially. In the inferior corneal endothelium cell density decreased to 45-50% of that in normals. Histologically, vacuoles in the iris and chamber angle were found inferiorly after 4 weeks. Chamber angle closures were present between 5 and 7 o'clock in those eyes where the PFC liquids had been removed after 2 and 4 weeks. Eyes with 0.025 cc PFD or PFP droplets showed vacuolization of the inferior trabecular meshwork 8 weeks postoperatively that was comparable with eyes which had a 50% aqueous replacement for 4 weeks. Control eyes remained unchanged in all aspects. CONCLUSION: Anterior segment damage caused by PFC liquids is a contact-dependent effect seen in the early observation period. Experimentally there was no difference between the products used or between 2 and 4 weeks' duration of the tamponade. PMID- 10379613 TI - Effects of systemically applied allopurinol and prednisolone on experimental autoimmune uveitis. AB - PURPOSE: To compare the effects of allopurinol to those of prednisolone on the oxidative tissue damage and inflammatory response in experimental autoimmune uveitis (EAU). METHODS: Experiments were performed using 27 male Lewis rats. EAU was induced by means of crude retina extract, Freund's adjuvant and pertussis toxin. One group of animals served as controls and two groups were treated systemically, one with allopurinol and one with prednisolone. At the end of the experiments lipid peroxides (LPO), myeloperoxidase activity (MPO), and histological changes were determined in the retinal tissue. LPO were measured by two different methods [thiobarbituric acid reactive substances (TBARS) and malondialdehyde-like substances]. RESULTS: Allopurinol led to a significant reduction in LPO and MPO levels. The steroid treatment also resulted in a significant reduction in MPO activity but LPO were significantly reduced only when measured as TBARS. Histological changes were significantly reduced by allopurinol only. DISCUSSION: Allopurinol is more effective than prednisolone in treating EAU. Its efficacy can be explained by the antioxidative/antiinflammatory and probably immunological action. The antiinflammatory effects of prednisolone are not sufficient to reduce the tissue damage. Allopurinol promises to be a useful alternative to steroids in the treatment of uveitis. PMID- 10379614 TI - Morphologic and genetic analysis of retinal angioma associated with massive gliosis in a patient with von Hippel-Lindau disease. AB - We report morphologic and genetic analysis of bilateral retinal angiomas in a 35 year-old patient with von Hippel-Lindau (VHL) disease. Enucleation of both eyes revealed extensive intraocular tumor. Whereas the right eye demonstrated large amounts of retinal angioma tissue, the left eye showed small areas of retinal angioma associated with massive diffuse retinal gliosis. Genetic analysis of the angioma showed allelic deletion of the VHL gene locus, suggesting that the origin of the angiomas was directly related to the patient's underlying VHL disease. Genetic analysis of the pleomorphic glial proliferation showed no allelic VHL gene deletion, which is consistent with the assessment that the glial component represents a reactive process. Apoptosis detected by TUNEL revealed lack of DNA fragmentation in the angioma; in contrast, many positive signals were found in the massive gliosis. We confirmed that the abnormal VHL genes were located in the "stromal" cells of the retinal angioma. Massive gliosis in VHL disease is a true reactive retinal gliosis. PMID- 10379616 TI - Loosening of single versus double running sutures in penetrating keratoplasty for keratoconus. AB - PURPOSE: Purpose of the study was to evaluate single versus double running sutures in penetrating keratoplasty for keratoconus with respect to suture loosening. METHODS: Eighty-eight patients were consecutively operated for keratoconus by the same surgeon with the same surgical technique. For the first 45 patients, a single running 10-0 nylon suture was used. For the remaining 43 patients, double running 10-0 nylon sutures were taken. RESULTS: Suture loosening was observed significantly (P<0.001; Chi-Square test) more often in the patients with a single running suture (12/45=27%) than in the patients with double running sutures (0/43=0%). CONCLUSIONS: The results indicate that double running sutures in comparison to a single running suture may be helpful in preventing suture loosening in penetrating keratoplasty for keratoconus. PMID- 10379615 TI - Eosinophilic granuloma (Kimura's disease) of the orbit: a case report. AB - BACKGROUND: Eosinophilic granuloma of the soft tissue, Kimura's disease, is a benign slow-growing tumor that is manifested clinically by one or more inflammatory nodules involving mainly the face and scalp, but rarely the eye. CASE REPORT: The patient was a 32-year-old male with swelling of the left lower eyelid, marked peripheral blood eosinophilia and increased serum immunogloblin E. MRI revealed swelling of all rectus muscles of the left eye, but no tumor mass. Corticosteroid treatment reduced the swelling of the eyelid, but it recurred after corticosteroid was discontinued. Eight years later the patient returned with a complaint of increased swelling of the left lower eyelid. An elastic, nontender, soft tumor mass was palpable subcutaneously in the left lower eyelid extending into the orbit. MRI revealed a tumor mass in the left orbital space. The parotid gland was also swollen and palpable. Both tumors were resected surgically, and histopathological study revealed prominent proliferation of lymphoid follicles with germinal centers showing interfollicular infiltration by eosinophils. The pathological findings in the parotid gland were similar. The diagnosis was Kimura's disease. CONCLUSION: This patient is unique in that he had no tumor at the first examination, only swelling of the rectus muscles, and a tumor mass appeared many years later. Unilateral swelling of the rectus muscles may be one of the first signs of Kimura's disease. Not only tumor but also swelling of the rectus muscles limited ocular movement. PMID- 10379617 TI - Dark spots in late-phase indocyanine green angiographic studies in a patient with presumed ocular histoplasmosis syndrome. AB - PURPOSE: We analyzed indocyanine green (ICG) angiograms in a patient with presumed ocular histoplasmosis syndrome (POHS) complaining about "seeing spots" and decreased visual acuity in order to identify the pathologic process. PATIENTS AND METHODS: A 30-year-old caucasian man with clinical signs of POHS who had previously undergone laser photocoagulation for secondary choroidal neovascularization developed visual disturbances primarily in his temporal visual field. We performed fundus photography, fluorescein angiography and ICG angiography before, during and after the episode of visual disturbance. ICG angiographic findings were correlated to fundus photographs and fluorescein angiograms. RESULTS: Fundus examination, fluorescein angiograms and early-phase ICG angiograms were unremarkable at all time points. However, during the phase of visual disturbance, late-phase ICG angiographic study revealed hypofluorescent lesions in the area representing the visual disturbances. At 1 week follow-up, these hypofluorescent lesions were reduced in size and number; at 6 months follow up they had completely resolved. CONCLUSIONS: Late-phase ICG angiographic study can provide additional information in inflammatory retinal disease by virtue of identifying areas of choroidal alterations while standard diagnostic examination remain unremarkable. PMID- 10379618 TI - Lasting effect of repeated cocaine administration on acoustic and fear potentiated startle in rats. AB - RATIONALE: Following cocaine withdrawal, humans may experience an abstinence syndrome with high levels of anxiety. Studying anxious behavior in animals following repeated cocaine administration may help elucidate important variables that contribute to a withdrawal syndrome. OBJECTIVES: This study investigated whether repeated cocaine pre-exposure produced lasting increases in conditioned fear as measured by fear-potentiated startle responses in rats. METHODS: Startle was measured in response to 50 ms acoustic stimuli of 95, 105 and 115 dB. Cocaine (20 mg/kg, IP) or saline was administered for 7 days and after each injection rats were either placed in startle chambers for 30 min or returned to the home cage. After a 1-week cocaine-free period, most rats were given ten light footshock pairings in the startle chamber. Fear-potentiated startle was tested by presenting acoustic startle-eliciting stimuli of 95, 105 and 115 dB in the presence or absence of the light. Rats that were not fear conditioned received acoustic stimuli one week after 7 days of 20 mg/kg cocaine. RESULTS: Startle responses, both in the presence and absence of the light CS, were greater in fear conditioned rats that received cocaine pre-exposure in the startle chamber than in saline pre-exposed rats. Startle responses in the presence of the light CS were further augmented at 115 dB. In contrast, home-cage exposure to cocaine did not enhance startle responses. In rats that were not fear conditioned, cocaine pre-exposure reduced acoustic startle. CONCLUSIONS: Repeated cocaine pre-exposure can increase, decrease or not change acoustic startle depending on whether fear conditioning occurred and whether cocaine was given in the testing chamber. The data suggest that cocaine pre-exposure may act as a contextually conditioned occasion setting stimulus that can facilitate anxious behavior similar to the postulated human cocaine abstinence syndrome. PMID- 10379619 TI - Triazolam and zolpidem: effects on human memory and attentional processes. AB - RATIONALE: The imidazopyridine hypnotic zolpidem may produce less memory and cognitive impairment than classic benzodiazepines, due to its relatively low binding affinity for the benzodiazepine receptor subtypes found in areas of the brain which are involved in learning and memory. OBJECTIVES: The study was designed to compare the acute effects of single oral doses of zolpidem (5, 10, 20 mg/70 kg) and the benzodiazepine hypnotic triazolam (0.125, 0.25, and 0.5 mg/70 kg) on specific memory and attentional processes. METHODS: Drug effects on memory for target (i.e., focal) information and contextual information (i.e., peripheral details surrounding a target stimulus presentation) were evaluated using a source monitoring paradigm, and drug effects on selective attention mechanisms were evaluated using a negative priming paradigm, in 18 healthy volunteers in a double blind, placebo-controlled, crossover design. RESULTS: Triazolam and zolpidem produced strikingly similar dose-related effects on memory for target information. Both triazolam and zolpidem impaired subjects' ability to remember whether a word stimulus had been presented to them on the computer screen or whether they had been asked to generate the stimulus based on an antonym cue (memory for the origin of a stimulus, which is one type of contextual information). The results suggested that triazolam, but not zolpidem, impaired memory for the screen location of picture stimuli (spatial contextual information). Although both triazolam and zolpidem increased overall reaction time in the negative priming task, only triazolam increased the magnitude of negative priming relative to placebo. CONCLUSIONS: The observed differences between triazolam and zolpidem have implications for the cognitive and pharmacological mechanisms underlying drug-induced deficits in specific memory and attentional processes, as well for the cognitive and brain mechanisms underlying these processes. PMID- 10379620 TI - Interactions between neuroleptics and 5-HT(1A) ligands in preclinical behavioral models for antipsychotic and extrapyramidal effects. AB - RATIONALE: Combining neuroleptics with 5-HT1A ligands is thought to improve the preclinical profile of neuroleptics and may be of interest in the development of new compounds that have greater therapeutic potential and/or are better tolerated. OBJECTIVE: To examine 1) the ability of 5-HT1A ligands to alter the effects of neuroleptics in preclinical models for antipsychotic potential (hindlimb retraction time in the paw test) and extrapyramidal side-effects (forelimb retraction time in the paw test; catalepsy tests), 2) the role of intrinsic activity at 5-HT1A receptors in the modulatory effects of 5-HT1A ligands, and 3) the generality of the interactions across neuroleptics. METHODS: The effects of different doses of 5-HT1A ligands with intrinsic activity ranging from high (e.g., 8-OH-DPAT) to low (e.g., WAY 100135) administered together with a fixed, high dose of the neuroleptics haloperidol, risperidone, and tropapride were examined in the paw test and on catalepsy. RESULTS: Firstly, the 5-HT1A agonists 8-OH-DPAT and ipsapirone attenuated the extrapyramidal-like effects of haloperidol and risperidone more than their therapeutic-like effects; this was not observed for tropapride, where all of its effects were markedly attenuated. Secondly, neither the weak 5-HT1A agonist WAY 100135 nor the silent antagonist WAY 100635 attenuated the effects of neuroleptics. Thirdly, neuroleptics apparently differed in their sensitivity to interactions with 5-HT1A agonists inasmuch as 8-OH-DPAT and ipsapirone attenuated the effects of tropapride on hindlimb retraction times more than those of haloperidol or risperidone. CONCLUSIONS: The present data suggest that 5-HT1A agonists with intermediate or high, but not low, intrinsic activity may abolish the extrapyramidal effects of neuroleptics. Together with results of previous studies, it appears that 5-HT1A agonists alter the antipsychotic-like effects of neuroleptics, although this may depend on the neuroleptic studied. PMID- 10379621 TI - Neuroleptic influences on a lateralized behavioral bias in unoperated rats. AB - RATIONALE: Abnormalities of the ascending dopaminergic system have been associated with hemineglect, and evidence suggests that neuroleptic treatments normalize the right hemispatial attentional impairment that is sometimes observed in acutely psychotic schizophrenic patients. OBJECTIVE: A research program was initiated to develop an animal model of hemineglect-drug interactions. METHODS: Female rats were tested repeatedly on removing a "nuisance stimulus" - strips of surgical tape applied loosely to the forelimbs. Subjects were assigned to groups dependent upon showing a behavioral orientation preference (BOP) during pre-drug baseline tests. Animals representing left-BOP, right-BOP and no preference continued to be tested during chronic exposure either to a dopamine antagonist (0.05 mg haloperidol/kg) or agonist (1.0 mg d-amphetamine/kg) or vehicle only. RESULTS: Three weeks of haloperidol treatments to rats only with an initial right BOP gradually eliminated the response bias, as revealed by declines in choosing the right forepaw and the latencies between attending to the two forepaws. CONCLUSION: The results recommend right BOP female rats as a simple, non-invasive behavioral animal model for evaluating and comparing neuroleptic medications. PMID- 10379622 TI - A double blind study of the effects of smoking on heart rate: is there tachyphylaxis? AB - RATIONALE: Smoking following overnight abstention reliably increases heart rate (HR), an effect due to nicotine absorption. The effect of subsequent cigarettes on HR is less than that associated with the first cigarette of the day, an indication of tachyphylaxis (acute tolerance). To date, smoking/HR studies have not been conducted double-blind. Instead, control conditions have included non smoking or some type of "sham" smoking (puffing on an unlit cigarette or a straw). OBJECTIVE: We investigated the HR response to smoking and its time course using double-blind methodology. METHODS: HR was recorded in overnight-abstaining participants before and after smoking the first, second and third cigarette of the day (40 min between each cigarette) in two sessions. The experimental manipulation involved replacing the second cigarette of one session with a very low nicotine-yield cigarette (0.05 mg; FTC method) compared with the other five cigarettes (1.1-mg nicotine yield). RESULTS: Smoking increased HR by 15, 8 and 7 beats/min (bpm) in the session where all three cigarettes had the higher yield. The comparable values for the session in which the second cigarette had the lower yield were 15, -1 and 11 bpm. CONCLUSIONS: In the session where all three cigarettes had the higher yield, larger increase in HR after smoking the first than the second or third cigarettes indicates tachyphylaxis. The HR response in the other session was smaller for the third cigarette than the first cigarette, indicating that a period greater than 80 min would be needed before the HR response was fully restored. PMID- 10379623 TI - Naltrexone and beta-funaltrexamine antagonism of the antinociceptive and response rate-decreasing effects of morphine, dezocine, and d-propoxyphene. AB - RATIONALE: Patterns of competitive and insurmountable antagonism provide important data to guide the classification and characterization of different types of opioid agonists as well as infer the mechanism of action for agonists. OBJECTIVE: Experiments with the competitive antagonist, naltrexone, and the insurmountable antagonist, beta-funaltrexamine (beta-FNA), were conducted to determine whether the antinociceptive and rate-decreasing effects of the opioid agonists dezocine and d-propoxyphene are 1) mediated through muu opioid receptors in rats, and 2) differ from morphine with respect to relative efficacy. METHODS: The rat tail-withdrawal assay was used to measure antinociception and a fixed ratio 20 (FR20) schedule of food delivery was used to measure rate suppression. RESULTS: Naltrexone (0.01-1.0 mg/kg) was approximately equipotent as an antagonist of the antinociceptive and rate-decreasing effects of both morphine and dezocine and as an antagonist of the antinociceptive effects of d propoxyphene. Naltrexone failed to block the rate-decreasing effects of d propoxyphene. beta-FNA (5 and 10 mg/kg) also antagonized the antinociceptive and rate-decreasing effects of morphine and dezocine as well as the antinociceptive effects of d-propoxyphene. beta-FNA failed to produce a dose-dependent antagonism of the rate-decreasing effects of d-propoxyphene. CONCLUSIONS: These data suggest that the antinociceptive effects of morphine, dezocine, and d-propoxyphene and the rate-decreasing effects of morphine and dezocine are mediated through mu opioid receptors. Overall, high doses of beta-FNA produced a greater degree of antagonism of the behavioral effects of dezocine than morphine or d-propoxyphene, confirming other reports that dezocine is a lower efficacy agonist than morphine. Additionally, the degree of antagonism produced by beta-FNA was greater for the antinociceptive effects of all three compounds than for the rate-decreasing effects. PMID- 10379624 TI - Modulation of behaviour on trials 1 and 2 in the elevated plus-maze test of anxiety after systemic and hippocampal administration of nicotine. AB - RATIONALE: The elevated plus-maze provides a test situation in which distinctive states of anxiety are elicited on trials 1 and 2 and the dorsal hippocampus has previously been shown to mediate the anxiogenic effects of (-)-nicotine in the social interaction test. OBJECTIVE: To determine the effects of a wide dose range of (-)-nicotine on trial 1 and 2 in the plus-maze after systemic administration and whether the dorsal hippocampus is a site mediating the anxiogenic effect of nicotine. METHODS: (-)-Nicotine (0.001, 0.005, 0.01, 0.05, 0.1, 0.5 and 1 mg/kg) was injected IP 30 min before testing for 5 min in the plus-maze. Rats receiving dorsal hippocampal infusions received bilateral infusions of 0.5 microl of artificial CSF or (-)-nicotine (0.1, 1, 4 or 8 microg). The needle was left in place for 50 s after injection and testing took place 3 min later. Rats tested on trial 1 were naive to the plus-maze, those tested on trial 2 had received a previous 5-min undrugged exposure to the maze 48 h earlier. RESULTS: Low doses of (-)-nicotine (0.001, 0.005, 0.01, 0.05 and 0.1 mg/kg, IP) were without effect on either trial, but higher doses (0.5 and 1 mg/kg, IP) had anxiogenic effects on both trials, as shown by decreases in percentage time spent and percentage entries onto the open arms. Infusion of (-)-nicotine (0.1, 1, 4 and 8 microg) bilaterally into the dorsal hippocampus was without effect on trial 1, but 1 microg had an anxiolytic effect on trial 2, shown by an increased percentage time spent on the open arms. CONCLUSIONS: The results on both trials in the plus-maze after systemic administration of nicotine add to previous reports from the social interaction test that high doses of nicotine have anxiogenic effects. However, the effects of nicotine in the dorsal hippocampus are different in all three anxiety tests (anxiogenic in social interaction, ineffective on trial 1, anxiolytic on trial 2) showing that nicotinic cholinergic control in this brain region may vary depending on the state and/or type of anxiety generated by the test. The brain region(s) underlying the anxiogenic effects of IP nicotine on both trials in the plus-maze remain to be identified. PMID- 10379625 TI - Effects of acute and repeated dose administration of caffeine and pentoxifylline on diazepam-induced mouse behavior in the hole-board test. AB - RATIONALE: The behavioral effects of methyl xanthines and their interactions with benzodiazepines have not been clearly established in animal models of anxiety. OBJECTIVE: The present study extended the previous studies to determine the effects of acute and repeated administration of caffeine, a non-specific phosphodiesterase (PDE) inhibitor and pentoxyfylline, a specific type-4 phosphodiesterase (PDE4) inhibitor on (1) baseline anxiety-like behavior and (2) the response to an acute challenge with diazepam on anxiety-like behavior in the hole-board test. METHODS: Mice were observed for the number of head-dips they made into the holes of the hole-board apparatus during a 5-min period, starting 30 min after acute (20 mg/kg) and repeated oral dose (20 mg/kg, twice a day for 4 days) administration of caffeine and pentoxifylline. In separate experiments, the response to an acute challenge with graded doses of diazepam (0.375 3 mg/kg, SC) was observed in naive mice or mice on acute and repeated dose regimen with methyl xanthines. RESULTS: Mice on acute but not after repeated dose regimen demonstrated a significantly increased number of hole-dips, indicating an anxiolytic-like effect of methylxanthines. Diazepam at the lower doses (0.375 and 0.75 mg/kg) but not at the highest doses (1.5 and 3 mg/kg) examined produced a significant anxiolytic-like effect. After an acute dose exposure of mice to caffeine and pentoxifylline, a rightward shift in the dose-response curve of diazepam was observed and particularly at 1.5 mg/kg dose, the net effect of diazepam was significantly enhanced which was, however, impaired upon repeated administration, more so with caffeine than with pentoxifylline. CONCLUSIONS: It is concluded that the xanthine drugs exert anxiolytic-like activity similar to diazepam in the hole-board test. In addition, they seem to modulate the anxiolytic effects of diazepam after both acute and repeated administration, probably as a result of an endogenous adenosinergic mechanism which may have therapeutic significance. PMID- 10379626 TI - Behavioural analysis of the acute and chronic effects of MDMA treatment in the rat. AB - RATIONALE: A variety of animal models have shown MDMA (3,4 methylenedioxymethamphetamine) to be a selective 5-HT neurotoxin, though little is known of the long-term behavioural effects of the pathophysiology. The widespread recreational use of MDMA thus raises concerns over the long-term functional sequelae in humans. OBJECTIVE: This study was designed to explore both the acute- and post-treatment consequences of a 3-day neurotoxic exposure to MDMA in the rat, using a variety of behavioural paradigms. METHODS: Following training to pretreatment performance criteria, animals were treated twice daily with ascending doses of MDMA (10, 15, 20 mg/kg) over 3 days. Body temperature, locomotor activity, skilled paw-reaching ability and performance of the delayed non-match to place (DNMTP) procedure was assessed daily during this period and on an intermittent schedule over the following 16 days. Finally, post mortem biochemical analyses of [3H] citalopram binding and monoamine levels were performed. RESULTS: During the MDMA treatment period, an acute 5-HT-like syndrome was observed which showed evidence of tolerance. Once drug treatment ceased the syndrome abated completely. During the post-treatment phase, a selective, delay dependent, deficit in DNMTP performance developed. Post-mortem analysis confirmed reductions in markers of 5-HT function, in cortex, hippocampus and striatum. CONCLUSIONS: These results confirm that acutely MDMA exposure elicits a classical 5-HT syndrome. In the long-term, exposure results in 5-HT neurotoxicity and a lasting cognitive impairment. These results have significant implications for the prediction that use of MDMA in humans could have deleterious long-term neuropsychological/psychiatric consequences. PMID- 10379627 TI - Sex differences in the acquisition of intravenously self-administered cocaine and heroin in rats. AB - RATIONALE: Despite numerous reports that male and female animals differ in behavioral responses to drugs, few studies have investigated sex differences in drug-reinforced behavior. OBJECTIVES: Acquisition of IV cocaine and heroin self administration was compared in 20 female and 22 male Wistar rats. METHODS: An autoshaping procedure was used to train rats to press a lever that resulted in either a 0.2 mg/kg infusion of cocaine or a 0.015 mg/kg infusion of heroin under a fixed-ratio 1 (FR 1) schedule. Daily sessions consisted of six 1-h autoshaping components followed by a 6-h self-administration component. During each autoshaping component, a retractable lever briefly (15 s) extended into the test chamber on a random interval schedule with a mean of either 90 s (cocaine groups) or 480 s (heroin groups) and either ten (cocaine groups) or five (heroin groups) computer-automated infusions were delivered each hour. During each 6-h self administration component, the lever remained extended and each response on the lever resulted in an infusion of either cocaine (0.2 mg/kg) or heroin (0.015 mg/kg). The criterion for acquisition of cocaine self-administration was a mean of at least 100 infusions and the criterion for heroin self-administration was a mean of at least 20 infusions during the self-administration component over five consecutive sessions. RESULTS: Female rats acquired both cocaine and heroin self administration more rapidly than males. Acquisition of cocaine self administration occurred in a greater percentage of female rats compared to males. Female rats self-administered more cocaine than males after acquisition criteria had been met. CONCLUSIONS: These findings indicate that female rats were more vulnerable than males to the acquisition of cocaine and heroin self administration under the conditions of the present experiment. PMID- 10379628 TI - Cyanamide reduces brain catalase and ethanol-induced locomotor activity: is there a functional link? AB - The present study was designed in an attempt to assess a previously suggested role of brain catalase activity in ethanol-induced behaviour by examining ethanol induced locomotor activity in cyanamide-treated mice. Mice were pretreated with IP injections of the catalase inhibitor cyanamide (3.75, 7.5, 15, 30 or 45 mg/kg) or saline. Following this treatment, animals in each group received IP injections of ethanol (0.0, 1.6, 2.4 or 3.2 g/kg) and locomotion was recorded. Several time intervals (0, 5, 10, 15, 20 or 25 h) between the two treatments were also evaluated. Results indicated that cyanamide administration produced a dose dependent decrease in ethanol-induced locomotor activity that depends on the time between treatments. However, cyanamide did not change spontaneous or d amphetamine-induced locomotor activity. Moreover, an additive effect of cyanamide and another brain catalase inhibitor, 3-amino-1,2,4-triazole (AT), on the reduction of ethanol-induced locomotor activity was observed. Perfused brain homogenates of mice treated with cyanamide, AT or cyanamide+AT showed a significant reduction of brain catalase activity. The dose and time patterns of both effects were closely related and a significant correlation between them was obtained. These results suggest that cyanamide could reduce locomotor activity through its inhibition of brain catalase, giving further support to the notion that brain catalase may be an important regulator of some ethanol-induced behavioural effects. PMID- 10379629 TI - The dopamine D3 antagonist U-99194A maleate increases social behaviors of isolation-induced aggressive male mice. AB - RATIONALE: Blockade of D1/D2 dopamine receptors produce an antiaggressive action commonly associated with an impairment of other motor behaviors. The D3 receptor seems to present opposite actions to the D1 and D2, since the blockade of this receptor produces stimulation of motor activity which has been associated with an increase in dopamine neurotransmission. OBJECTIVE: In this work, the action of the dopamine D3 antagonist U-99194a maleate on locomotor activity and in a social interaction test in male mice was evaluated. METHODS: Animals isolated during 30 days were treated with U-99194a maleate (20-40 mg/kg) or saline and locomotor activity was measured 20 min after drug administration. The behavioral interaction test was performed afterwards, between the experimental isolated animal and a standard opponent. RESULTS: The higher dose used produces a significant decrease in spontaneous motor activity and presents an antiaggressive action without impairment of other behaviors, such as nonsocial exploration or immobility. At all doses tested, U-99194a maleate significantly increases social investigation. CONCLUSIONS: Our results give support to the hypothesis that the D3 receptor could play a role in emotional behaviors. PMID- 10379630 TI - A comparison of repaglinide and glibenclamide in the treatment of type 2 diabetic patients previously treated with sulphonylureas. AB - OBJECTIVE: To compare the efficacy and safety of repaglinide, a novel oral prandial glucose regulator, with that of glibenclamide, an oral hypoglycaemic agent, in the treatment of patients with type 2 diabetes. METHODS: This was a 14 week, double-blind, parallel-group trail in which a total of 195 type 2 diabetic patients treated with oral hypoglycaemic agents were randomized to receive either repaglinide, administered preprandially three times daily, or glibenclamide, given preprandially once or twice daily, as per manufacturer's recommendations. RESULTS: By the end of the study, the 2-h postprandial blood glucose values were lower in the repaglinide group than in the glibenclamide group, with the difference approaching statistical significance (repaglinide, 8.1 (0.6) mol x l( 1) vs glibenclamide, 9.1 (0.6)mmol x l(-1); P = 0.07). There was no statistically significant difference in the mean blood glucose level at the end of the study between the two groups (repaglinide, 7.1 (0.5) mmol x l(-1) vs glibenclamide, 7.4 (0.5) mmol x l(-1); P = 0.42), and baseline HbA1c values had decreased to the same degree in both the repaglinide [7.8% (0.1%) to 7.5% 0.1%)] and the glibenclamide groups [8.0 (0.1%) to 7.6 (0.1%)]. There are no significant differences between the repaglinide and glibenclamide treatment groups in the levels of fasting blood glucose, fructosamine, fasting C-peptide, insulin and proinsulin. Neither treatment group showed any clinically significant changes in blood lipid profiles. Repaglinide and glibenclamide were both well tolerated. No significant differences were observed between the two treatment groups with respect to adverse events, including hypoglycaemic episodes and weight change. No accumulation of repaglinide was apparent during the maintenance period. CONCLUSION: Repaglinide is as well tolerated as glibenclamide and is equally effective in the management of type 2 diabetes. Repaglinide may, however, offer an improvement in postprandial blood glucose control compared with glibenclamide, thereby helping to reduce the relative long-term risk of diabetic complications. PMID- 10379631 TI - Neuromuscular blocking characteristics of vecuronium after tubocurarine-induced "fade". An experimental double-blind clinical study. AB - OBJECTIVE: The fade in train-of-four (TOF) monitoring is considered to be due to blocking of the prejunctional nicotinic acetylcholine receptors (AchRs). During onset of the neuromuscular block (NMB) tubocurarine (TC) causes more fade in the TOF responses than vecuronium (VEC). Therefore we wanted to investigate whether onset or duration of action of VEC or TC would be improved with a priming dose of an agent with different prejunctional activity. METHODS: The rates of NMB were measured following priming doses of 0.15 mg x kg(-1) of TC and 0.015 mg x kg(-1) of VEC with 6 min priming time. The individual time course of action of 0.6 mg x kg(-1) of TC (1.13 x ED 95) and 0.1-0.2 mg x kg(-1) of VEC (1.75-3.5 x ED95) were examined with a priming dose of the same agent or the other agent, by measurement of changes in the evoked compound EMG from the hypothenar muscle. RESULTS: Priming doses of TC decreased mean TOF ratio to 67% [95% confidence interval (CI) = 56-78] during priming time, which was significantly lower than after priming with VEC 87% (76-97; P < 0.001). Despite the higher TOF ratio, the priming dose of VEC accelerated the onset time of intubation dose of TC more than the priming dose of TC (P = 0.0018). Priming with TC prolonged the duration of VEC-induced NMB by 35-70 min compared with priming with VEC, which means that a small priming dose of TC changes VEC from a muscle relaxant with intermediate action to a long acting agent. CONCLUSION: Priming with TC caused a lower TOF ratio; however, priming with TC did not accelerate the onset time of either agent as much as priming with VEC. It appears that potentiation of NMB after combination of VEC and TC is not dependent on "fade" receptors. PMID- 10379632 TI - CYP2D6 and CYP2C19 activity in a large population of Dutch healthy volunteers: indications for oral contraceptive-related gender differences. AB - OBJECTIVE: We examined a large database containing results on CYP2D6 and CYP2C19 activity of 4301 Dutch volunteers phenotyped in the context of various clinical pharmacology studies. METHODS: The subjects were given 22 mg dextromethorphan, 100 mg mephenytoin and 200 mg caffeine. For CYP2D6, the dextromethorphan/dextrorphan metabolic ratios in urine samples taken for a subsequent 8 h were used. Dextromethorphan and dextrorphan were quantified by reversed-phase high performance liquid chromatography. For CYP2C19 similarly obtained (R)-mephenytoin and (S)-mephenytoin ratios were used. (S)-mephenytoin and (R)-mephenytoin were analysed and quantified by enantioselective capillary gas chromatography. In addition, CYP2C19 poor metabolizer (PM) subjects were reanalysed after acidic pre-treatment of urine samples to confirm the PM status. RESULTS: The investigated population mainly comprised Caucasian (98.9%) males (68%). The age ranged from 18 to 82 years. For CYP2D6, it was found that 8.0% of the subjects were PMs. The average metabolic ratio was 0.014 (0.033) for subjects who showed extensive metabolizing activity (EM) and 5.4 (7.6) for PM subjects. For CYP2C19, it was found that 1.8% of the subjects were PMs. The metabolic ratio was 0.162 (0.124) for EM subjects and 1.076 (0.040) for PM subjects. Within the EM group the metabolic ratio in females was significantly lower for CYP2D6 (-20%) and significantly higher for CYP2C19 (+40%) compared with males. For PMs there was no such difference for CYP2D6 (P = 0.79) or CYP2C19 (P = 0.20). Oral contraceptive (OC) use significantly decreased the CYP2C19 activity by 68% for mephenytoin as compared to non-OC using females. CONCLUSIONS: For CYP2D6, the PM incidence (8.0%) is in accordance with literature data. The CYP2C19, PM incidence (1.8%) is low compared to reports from other European countries. For mephenytoin, the acidification procedure has been shown to be very important for the confirmation of CYP2C19 PMs. In EM females compared to EM males, CYP2D6 activity is increased and CYP2C19 activity is reduced. For CYP2C19 in particular this reduction is substantial and most pronounced in the age range from 18 to 40 years. For CYP2C19, the reduced activity is associated with the use of oral contraceptives. PMID- 10379634 TI - The effect of four different antihypertensive medications on cardiovascular regulation in hypertensive sleep apneic patients--assessment by spectral analysis of heart rate and blood pressure variability. AB - OBJECTIVE: To study the effect of antihypertensive medications on autonomic nervous system in patients with hypertension and sleep apnea syndrome using frequency domain measures of heart rate and blood pressure variabilities. METHODS: The beta-receptor blocking agent atenolol (50 mg), the calcium antagonist isradipine SRO (2.5 mg), the diuretic hydrochlorothiazide (25 mg) and the ACE inhibitor spirapril (6 mg) once daily were given in a double-blind crossover schedule for 8 weeks. Cardiovascular autonomic control was assessed using frequency domain measures of heart rate variability during the spontaneous and controlled breathing tests. During orthostatic maneuver and cold pressor test the blood pressure variability analysis also was performed. RESULTS: In general, the responses of heart rate and blood pressure variabilities were abnormal in the patients with arterial hypertension and sleep apnea syndrome compared to reference data. Of the four drugs, only atenolol effected heart rate and blood pressure variabilities as it shifted the autonomic regulation to the vagal direction. Other antihypertensive drugs did not change any parameter of heart rate or blood pressure variabilities. CONCLUSION: The short-term treatment with atenolol in patients with arterial hypertension and sleep apnea syndrome is associated with normalization of autonomic nervous control judged by heart rate and blood pressure variability. Thus, beta-receptor blockade may have adjunctive beneficial effects beyond blood pressure reduction in these patients. However, the long-term effects of blood pressure reduction on autonomic nervous control remain to be studied. PMID- 10379633 TI - Efficacy and tolerability of moexipril and nitrendipine in postmenopausal women with hypertension. MADAM study group. Moexipril as Antihypertensive Drug After Menopause. AB - OBJECTIVE: The aim of this study was to compare the efficacy and tolerability of the new angiotensin-converting enzyme (ACE) inhibitor moexipril and the calcium antagonist nitrendipine in postmenopausal women with mild to moderate hypertension. METHODS: After a 4-week placebo run-in period, 93 postmenopausal women (age range 44-70 years) with primary hypertension were randomized to receive moexipril 15 mg once daily or nitrendipine 20 mg once daily for 8 weeks. The mean sitting systolic (SSBP) and sitting diastolic blood pressures (SDBP) at baseline were 161.3/103.0 mmHg in the moexipril group, and 162.2/102.3 mmHg in the nitrendipine group. RESULTS: After the 8 weeks of treatment, the SSBP/SDBP reductions were -21.2/-15.2 mmHg in the moexipril group and -18.2/-13.6 mmHg in the nitrendipine group. Blood pressure responses were adequate in 82.2% of the moexipril-treated patients and in 80.9% in the nitrendipine-treated group. Adverse events were more frequent with nitrendipine than with moexipril. The most common adverse events in the nitrendipine group were headache (23.4%), flushing (21.3%) and ankle oedema (14.9%). In the moexipril group the most common adverse event was cough (8.9%). CONCLUSION: The results of the study suggest that moexipril and nitrendipine are equieffective in the given dosages. In the patient population of postmenopausal women, the ACE inhibitor moexipril appears to have an advantage over the calcium antagonist nitrendipine with regard to tolerability. PMID- 10379635 TI - Comparative pharmacokinetics and pharmacodynamics of the novel rapid-acting insulin analogue, insulin aspart, in healthy volunteers. AB - OBJECTIVE: The pharmacokinetics of a new insulin analogue, insulin aspart, were compared with unmodified human insulin in a double-blind crossover study of 25 fasting healthy men following a single subcutaneous dose. METHODS: Either insulin aspart or human insulin, 0.1 U x kg-body-weight(-1), was injected subcutaneously and followed by determination of 8-h profiles of serum insulin and plasma glucose concentrations. RESULTS: The absorption of insulin aspart was, on average, more than twice as fast and reached levels more than twice as high compared with human insulin [tmax(ins) of 52 (23) vs 145 (93) min, P < 0.0001; and Cmax(ins) of 41 (11) vs 18 (4) mU x l(-1), P < 0.0001; mean with (SD)]. However, total bioavailability did not differ between the insulins, and thus the mean residence time was significantly shorter for insulin aspart [MRT(ins) of 149 (26) vs 217 (30) min, P < 0.0001]. Plasma glucose (PG) fell more than twice as rapidly [tmin(PG) of 94 (45) vs 226 (120) min, P < 0.0001], to a greater extent [Cmin(PG) 2.1 (0.6) vs 1.4 (0.4) mmol x l(-1), P < 0.0001], and for a shorter duration with insulin aspart than with human insulin. CONCLUSION: With improved subcutaneous absorption characteristics, the insulin aspart concentration-time profile resembles physiological meal-stimulated insulin release more closely than that of unmodified human insulin. This significantly alters the pharmacodynamic response in an advantageous manner in the meal-related treatment of diabetes mellitus. PMID- 10379636 TI - Renal handling of drugs in the healthy elderly. Creatinine clearance underestimates renal function and pharmacokinetics remain virtually unchanged. AB - OBJECTIVE: It is commonly assumed that renal function, and in parallel the excretion of drugs, is considerably reduced in the elderly. Endogenous creatinine clearance or indirect estimates of this parameter are generally recommended for adapting drug dosage. The present study evaluates the validity of both assumptions. METHODS: We compared pharmacokinetics (and pharmacodynamics) of 50 mg atenolol, 800 mg piracetam and 25 mg hydrochlorothiazide plus 50 mg triamterene in ten healthy young [25 (2) years] and 11 healthy elderly subjects [68 (5) years]. Inulin (Cin) and para-aminohippurate [PAH (CPAH)] clearance (infusion clearance technique), endogenous (C(Cr)) and calculated (Cockroft Gault) creatinine clearance, analysis of drugs and their metabolites (HPLC), were performed. Renal haemodynamics and the pharmacokinetics of beta-adrenergic blocking agent, diuretics and the nootropic agent piracetam, respectively, were measured on separate days. RESULTS: Cin was significantly (P < 0.01) lower in the healthy elderly subjects [104 (12) vs 120 (14) ml x min(-2) x 1.73 m(-2) in the young], but remained within the normal range (> 90 ml x min(-2) x 1.73 m(-2)). In contrast, C(Cr) was even lower in healthy elderly subjects [95 (24) vs 121 (20) ml x min(-1) in the young], and the Cockroft-Gault clearance underestimated true glomerular filtration rate (GFR) even more seriously [74 (17) vs 122 (16) ml min( 1)]. For atenolol the mean area under the curve (AUC) was similar in both groups [3.16 (0.48) microg x h(-1) x ml(-1) in the elderly vs 3.01 (0.30) in the young], as was the mean maximal plasma concentration [0.42 (0.07) vs 0.44 (0.06) microg x ml(-1)], but the proportion of the drug excreted in urine was marginally (P < 0.025) lower in the elderly. Similar results were obtained for hydrochlorothiazide, whereas no marked differences between the groups were found for triamterene and its metabolite. Furthermore, the pharmacodynamic action of diuretics was not significantly altered in the elderly. CONCLUSIONS: The true GFR of the healthy elderly remains within the normal range and is underestimated by creatinine clearance and more so by its surrogate (Cockroft-Gault clearance). In parallel, pharmacokinetics of renally excreted drugs are not affected in the healthy elderly to a clinically significant extent. For drugs with a narrow therapeutic window, indirect estimates of GFR appear to be an unreliable means for calculating correct dosage in the elderly. PMID- 10379637 TI - Development of a general method of limited sampling for the determination of AUC for a drug that displays two-compartment pharmacokinetics. AB - OBJECTIVES: To develop a method of limited sampling that would enable accurate estimation of the area under the concentration time curve (AUC) when using the log trapezoidal method. METHODS: A series of datasets were simulated. Each dataset comprised 1000 subjects. Each subject was "administered" an intravenous bolus dose of a drug that displays two compartment pharmacokinetics. In the first series of simulations, a variety of combinations of the number of sampling times (K) and number of replicate measurements (R) at each of these times were tested, where K x R = 12 (i.e. N = 12). The times that each of the K samples were taken were chosen to be those that divided the AUC into K - 1 trapezoids of equal area. The concentration-time curves were estimated based on a priori estimates of the population parameters. The best combination of K and R was tested under various conditions of parameter variability and assay variability. The combinations were compared with a conventional sampling strategy, where N = 12, K = 12 (R = 1). RESULTS: The combination K = 4 and R = 3 proved to be the "best". It had similar accuracy to the conventional method. The best limited sampling combination was superior to the conventional method when assay variability was high (CV= 30%), was similar when assay variability was 15%, but the conventional method became statistically superior when assay variability was 7.5% or less. The accuracy of the best limited sampling combination was inversely related to the parameter variability. If K was set to 4 and R allowed to increase to 6 (i.e. N is not equal to 12), there was no further gain in accuracy. CONCLUSION: The proposed method of limited sampling is at least as accurate as the conventional intensive sampling technique, but more efficient in terms of sampling. PMID- 10379638 TI - Absence of interaction between erythromycin and a single dose of clozapine. AB - OBJECTIVE: To study the suggested pharmacokinetic interaction between erythromycin, a strong inhibitor of CYP3A4, and clozapine. METHODS: Twelve healthy male volunteers received a single dose of 12.5 mg of clozapine alone or in combination with a daily dose of 1500 mg erythromycin in a randomised crossover study. Clozapine and its metabolites clozapine-N-oxide and desmethyl clozapine were measured in serum samples which were collected during a 48 h period and in a sample of the urine secreted over the interval 0-12 h. RESULTS: There were no significant differences in mean area under the serum concentration time curves (1348 (633) nmol h x l(-1) in the control phase and 1180 (659) nmol h x l(-1) in the erythromycin phase), terminal halflives (19 (13) h and 15 (6) h, respectively), peak serum concentrations (92 (53) nmol x l(-1) and 77 (40) nmol x l(-1), respectively), time to peak serum concentrations (1.4 (0.7) h and 1.5 (1.0) h, respectively) or apparent oral clearances of clozapine (34 (15) l x h( 1) and 46 (37) l x h(-1), respectively). There were no significant differences in partial metabolic clearances to clozapine-N-oxide (5.1 (3.6) l x h(-1) and 7.8 (9.4) l x h(-1), respectively) or to desmethyl-clozapine (1.5 (1.3) l x h(-1) and 1.8 (1.7) l x h(-1), respectively) or in renal clearances of clozapine (0.8 (0.5) l x h(-1) and 1.0 (0.7) l x h(-1), respectively) between the two phases. CONCLUSION: These results demonstrate that erythromycin at a clinically relevant dosage does not inhibit the metabolism of clozapine. Hence, CYP3A4 seems to be of minor importance in the disposition of clozapine in humans at least when clozapine is taken at a low single dose. PMID- 10379639 TI - Effect of sulfasalazine on cyclosporin blood concentration. PMID- 10379640 TI - Energy level of P+B- with respect to P* found from recombination fluorescence measurements in pheophytin-modified reaction centres. AB - Recent studies of reaction centres from Rhodobacter sphaeroides (R-26), in which bacteriopheophytins a were replaced by plant pheophytins a, have shown that at low temperature the excited state of primary electron donor P* is converted to the state P+B-(A) (where B(A) is a bacteriochlorophyll a monomer in branch A) which has a long lifetime (about 600 ps [8]). This allows the direct measurement of the free energy difference between P* and P+B-(A) using the temperature dependence of the recombination fluorescence from P+B-(A). The data show that P+B (A) is located below P* by 550+/-30 mV. Thus, the primary conversion of P* leads to the formation of P+B-(A) which is below P* in energy and is a real intermediate in electron transfer. PMID- 10379641 TI - Organization and role of the long-wave chlorophylls in the photosystem I of the Cyanobacterium spirulina. AB - The data on the organization and function of the photosystem I pigment-protein complexes of the cyanobacterium Spirulina and the characteristics of pigment antenna of the photosystem I monomeric and trimeric core complexes are presented and discussed. We proved that the photosystem I complexes in the cyanobacterial membrane pre-exist mainly as trimers, though both types of complexes contribute to the photosynthetic electron transport. In contrast to monomers, the antenna of the photosystem I trimeric complexes of Spirulina contains the extreme long-wave chlorophyll form absorbing at 735 nm and emitting at 760 nm (77 K). The intensity of fluorescence at 760 nm depends strongly on the P700 redox state: it is maximum with the reduced P700 and strongly decreased with the oxidized P700 which is the most efficient quencher of fluorescence at 760 nm. The energy absorbed by the extreme long-wave chlorophyll form is active in the photooxidation of P700 in the trimeric complex. The data obtained indicate that the long-wave form of chlorophyll originates from interaction of the chlorophyll molecules localized on monomeric subunits forming the photosystem I trimer. Kinetic analysis of the P700 photooxidation and light-induced quenching of fluorescence at 760 nm (77 K) allows the suggestion that the excess energy absorbed by the antenna monomeric subunits within the trimer migrates via the extreme long-wave chlorophyll to the P700 cation radical and is quenched, which prevents the photodestruction of the pigment-protein complex. PMID- 10379642 TI - Possible relationship of mechanisms of Mn2+ oxidation and formation of plant photosystem II manganese cluster. AB - The mechanisms of Mn2+ cation oxidation in alkaline, neutral and slightly acidic media were studied. In all cases, the Mn2+ oxidation resulted in the formation of the structure[see text]. The formal resemblance and differences in the Mn2O3 structure and Klein's model of the Mn cluster of PS II were noted. The necessity of the primary ligation of Mn2+ cations was discussed for both the decrease in the Mn2+ oxidation potential and the stability of the Mn2O3 structure. It was supposed that Mn2O3 is an initial block for the assembly of the inorganic core of the photosynthetic water-oxidizing complex. PMID- 10379644 TI - The final stage of chlorophyll biosynthesis and formation of reaction centres of photosystems. AB - The survey reviews Russian studies of the final stage of chlorophyll biosynthesis and the role of A. A. Krasnovsky (Sr.) in the development of this direction. The current state of the problem is considered. The results of studies carried out by Krasnovsky's followers at Moscow State University are summarized. Schemes of the pathways of chlorophyll and pheophytin biosynthesis, biogenesis of pigment complexes of the two photosystems of photosynthesis proposed by those scientists are presented. PMID- 10379643 TI - Effect of deuteration and cryosolvents on the energy transduction in primary processes of photosynthesis. AB - Effects of cryosolvents and D2O/H2O substitution on the reaction centres (RCs) isolated from photosynthetic bacteria were studied with respect to the role of intra-protein hydrogen bonds in the primary photosynthetic electron transfer. As a result of such treatment of RCs, the charge separation rate between the photoactive bacteriochlorophyll (P2 dimer) and bacteriopheophytin and the rate of electron transfer to the primary quinone slowed down. The energy migration rate from bacteriopheophytin (BPheM), inactive in electron transport, to P2 decreased as well. Although cryosolvents can shift the redox potential of the photoactive pigment, there is no direct correlation between the P2 potential and the effects of these modifying agents on the photosynthetic process in RCs occurring with participation of P2. The removal of H subunit from the pigment-protein complex results in the pronounced weakening of the dimethyl sulfoxide modifying effects on the RC hydrogen bonds. The role of structural and dynamic state in the functioning of the photosynthetic bacterial RCs is analyzed. Relaxation processes in purple bacteria RCs accompanying the primary picosecond steps of energy transformation proceed with the participation of small proton-containing molecular groups in the immediate surroundings of electron transfer carriers. In this paper, we present results concerning mechanisms of primary photosynthetic steps, which were initiated by A. A. Krasnovsky and have been studied for several years at the Department of Biophysics. This paper is dedicated to the memory of our teacher Prof. A. A. Krasnovsky. PMID- 10379645 TI - Mechanisms of regulation and interplastid localization of chlorophyll biosynthesis. AB - The current concepts of chlorophyll biosynthesis, its interplastid localization, biosynthetic and biochemical heterogeneity, mechanisms of regulation of the key reactions, formation of 5-aminolevulinic acid and incorporation of magnesium into protoporphyrin IX, are reviewed. The literature and author's data demonstrate the existence of in vivo multienzyme systems synthesizing chlorophyll and its precursors as monovinyl and divinyl chemical species. Both types of the multienzyme systems synthesize 5-aminolevulinic acid and regulate this process independently. A hypothesis is considered that the function of the magnesium branch of chlorophyll biosynthesis in vivo is controlled by a mechanism through inhibition of the enzymes by their products because of the limitation of the binding sites for them in the membrane. An additional influence of light on the Mg-chelatase activity not only via the photosynthetic supply with ATP but also through the light-induced synthesis of the enzyme molecules de novo is described. Efficient energy migration from protoporphyrin IX and Mg-protoporphyrin IX (monomethyl ester) molecules to the protochlorophyllide active form detected by the author is discussed considering a close location of these pigments in plastid membranes and the enzymes participating in their formation. PMID- 10379646 TI - Free radicals in primary photobiological processes. AB - Formation of a semiquinone free radical derived from chlorophyll in the reaction of photoreduction has been discovered by A. A. Krasnovsky, Sr. in 1953. This review consider the results obtained in the author's laboratory, concerning the participation of free radicals in photochemical reactions under UV-irradiation of aromatic amino acids, proteins, and lipids, as well as in the reactions of chemiluminescence (CL) in the protein and chlorophyll-containing systems. Free radicals are the very first products of photochemical reactions in all systems studied. The back reactions of radicals are accompanied with photon emission. From the point of view of the molecular energetics, the radiativeless electronic transition in molecules is the most probable event, the transition triplet level is less probable, and the transition to the singlet excited level is virtually impossible. This may explain the low quantum yield of CL, similarity of CL and phosphorescence (rather than fluorescence) spectrum of the reaction products, low quantum yield of CL, and its high temperature coefficient. PMID- 10379647 TI - Singlet molecular oxygen in photobiochemical systems: IR phosphorescence studies. AB - Singlet molecular oxygen (1O2) is one of the most active intermediates involved in photosensitized oxygenation reactions in chemical and biological systems. Deactivation of singlet oxygen is accompanied by infrared phosphorescence (1270 nm) which is widely employed for 1O2 detection and study. This review considers techniques for phosphorescence detection, phosphorescence spectra, quantum yields and kinetics under laser excitation, the radiative and real 1O2 lifetimes in organic solvents and water, 1O2 quenching by biomolecules, and estimation of singlet oxygen lifetimes, diffusion lengths and phosphorescence quantum yields in blood plasma, cell cytoplasm, erythrocyte ghosts, retinal rod outer segments and chloroplast thylakoids. The experiments devoted to 1O2 phosphorescence detection in photosensitizer-containing living cells are discussed in detail. Information reviewed is important for understanding the mechanisms of photodestruction in biological systems and various applied problems of photobiology and photomedicine. PMID- 10379648 TI - The system of phytochromes: photobiophysics and photobiochemistry in vivo. AB - Phytochrome is a key photoregulation pigment in plants which determines the strategy of their development throughout their life cycle. The major achievement in the recent investigations of the pigment is the discovery of its structural and functional heterogeneity: existence of a family of phytochromes (phyA-phyE) differing by the apoprotein was demonstrated. We approach this problem by investigating the chromophore component of the pigment with the use of the developed method of in vivo low-temperature fluorescence spectroscopy of phytochrome. In etiolated plants, phytochrome fluorescence was detected and attributed to its red-light absorbing form (Pr) and the first photoproduct (lumi R), and a scheme of the photoreaction in phytochrome, a distinction of which is the activation barrier in the excited state, was put forward. It was found that the spectroscopic and photochemical characteristics of Pr depend on the plant species and phytochrome mutants and overexpressors used, on localization of the pigment in organs and tissues, plant age, effect of preillumination and other physiological factors. This variability of the parameters was interpreted as the existence of at least two phenomenological Pr populations, which differ by their spectroscopic characteristics and activation parameters of the Pr --> lumi-R photoreaction (in particular, by the extent of the Pr --> lumi-R photoconversion at low temperatures, gamma1): the longer-wavelength major and variable by its content in plant tissues Pr' with gamma1 = 0.5 and the shorter-wavelength minor relatively constant Pr" with gamma1 < or = 0.05. The analysis of the phytochrome mutants and overexpressors allows a conclusion that phytochrome A (phyA), which dominates in etiolated seedlings, is presented by two isoforms attributed to Pr' and Pr" (phyA' and phyA", respectively). Phytochrome B (phyB) accounts for less than 10% of the total phytochrome fluorescence and belongs to the Pr" type. It is also characterized by the relatively low extent of the Pr photoconversion into the far-red-light absorbing physiologically active phytochrome form, Pfr. Fluorescence of the minor phytochromes (phyC-phyE) is negligible. The recently discovered phytochrome of the cyanobacterium Synechocystis also belongs to the phenomenological Pr" type. PhyA' is a light-labile and soluble fraction, while phyA" is a relatively light-stable and, possibly, membrane (protein)-associated. Experiments with transgenic tobacco plants overexpressing full-length and C- and N-terminally truncated oat phytochrome A suggest that phyA' and phyA" might differ by the post-translational modification of the small N-terminal segment (amino acid residues 7-69) of the pigment. PhyA' is likely to be active in the de etiolation processes while phyA" together with phyB, in green plants as revealed by the experiments on transgenic potato plants and phytochrome mutants of Arabidopsis and pea with altered levels of phytochromes A and B and modified phenotypes. And finally, within phyA', there are three subpopulations which are, possibly, different conformers of the chromophore. Thus, there is a hierarchical system of phytochromes which include: (i) different phytochromes; (ii) their post translationally modified states and (iii) conformers within one molecular type. Its existence might be the rationale for the multiplicity of the photoregulation reactions in plants mediated by phytochrome. PMID- 10379649 TI - Ca2+ and intracellular signalling in plant cells: a role in phytochrome transduction. AB - The role of Ca2+ as a mediator and its participation in intracellular signaling in relation to phytochrome regulatory action are considered. Using the literature data, the non-random distribution of Ca2+ in the plant cell cytoplasm, the mechanisms of Ca2+ homeostasis and the function of putative messengers of Ca2+ are described. On the basis of the data obtained at the author's laboratory, a dual effect of phytochrome on the cytosolic Ca2+ concentration is demonstrated, i.e. phytochrome controls Ca2+ permeability of plasma membrane and induces transient changes in [Ca2+]cyt. Evidence is presented that the endoplasmic reticulum and vacuole are intracellular Ca2+ stores in the plant cell sensitive to phytochrome regulatory action. It is also demonstrated that phytochrome controls the metabolism of a key phosphoinositide in plant cell membranes. It stimulates the hydrolysis of phosphatidyl 4,5-bisphosphate, a precursor of inositol 1,4,5-trisphosphate, the potent mobilizer of Ca2+ in cytoplasm. Finally, the data on the close interactions between Ca2+ and other secondary messengers in the plant cell cytoplasm are presented. Using the transgenic tobacco expressing an apoprotein of Ca(2+)-sensitive photoprotein aequorin, evidence was obtained that cyclic mononucleotides (cGMP, cAMP) control the Ca2+ concentration in the cytoplasm testifying a crosstalk among Ca2+ and cyclic mononucleotides as messengers. PMID- 10379650 TI - Photoreactivation of the cytochrome oxidase complex with cyanide: the reaction of heme a3 photoreduction. AB - Electron transfer activity of isolated cytochrome oxidase inhibited by low concentrations of cyanide by 93-95% was shown to rise no less than three times under exposure to visible light. Irradiation with visible light was found to increase the rate of reduction of cytochrome oxidase heme groups in the presence of sodium dithionite. Based on these results, it is suggested that the modification of the catalytic and spectral characteristic of the cytochrome oxidase-cyanide complex is due to the photostimulation of the intramolecular electron transport at the interheme (heme a heme a3) transfer stage, i.e., is caused by photoreduction of the enzyme's heme a3-CN complex. PMID- 10379651 TI - Inorganic semiconductors as photosensitizers in biochemical redox reactions. AB - The results of studies of biochemical redox reactions photosensitized by inorganic semiconductor particles are reviewed. The mechanisms of hydrogen photoproduction, NAD+ or NADP+ photoreduction, CO2 photofixation and photosynthesis of organic and amino acids under the coupled action of TiO2, ZnO, CdS, ZnS and enzymes or bacterial cells are considered. Studies on the photocatalytic activity of ferritin, a protein containing microcrystals of hydrous ferric oxide, are described. The data on biosynthesis of cadmium sulfide by microorganisms and plants are analyzed. The possibility of the participation of inorganic semiconductors in photoprocesses in vivo is discussed. PMID- 10379652 TI - Separation and sensing based on molecular recognition using molecularly imprinted polymers. AB - Molecular recognition-based separation and sensing systems have received much attention in various fields because of their high selectivity for target molecules. Molecular imprinting has been recognized as a promising technique for the development of such systems, where the molecule to be recognized is added to a reaction mixture of a cross-linker(s), a solvent(s), and a functional monomer(s) that possesses a functional groups(s) capable of interacting with the target molecule. Binding sites in the resultant polymers involve functional groups originating from the added functional monomer(s), which can be constructed according to the shape and chemical properties of the target molecules. After removal of the target molecules, these molecularly imprinted complementary binding sites exhibit high selectivity and affinity for the template molecule. In this article, recent developments in molecularly imprinted polymers are described with their applications as separation media in liquid chromatography, capillary electrophoresis, solid-phase extraction, and membranes. Examples of binding assays and sensing systems using molecularly imprinted polymers are also presented. PMID- 10379653 TI - Purification and quantitation of tumor necrosis factor receptor immunoadhesin using a combination of immunoaffinity and reversed-phase chromatography. AB - The development of an automated, dual column assay to quantitate and recover the glycoprotein, tumor necrosis factor receptor immunoadhesin (TNFr-IgG) from monkey plasma, human serum, cell culture fluid and buffer samples is described. A combination of immunoaffinity and reversed-phase chromatographies are used. The targeted protein was captured using an anti-TNFr-1 monoclonal antibody immobilized on POROS resin. After non-specific adsorption had been reduced, the affinity column was placed in-line with a reversed-phase column and eluted with dilute acid. The reversed-phase column was subsequently eluted with an acetonitrile gradient and the TNFr-IgG collected and quantitated by comparison with peak areas of similarly treated standards. Detection was performed by measurement of absorbance at 214 nm. The dynamic range is from 0.5-15 microg total sample. Samples were quantitated and recovered from monkey and human pharmacokinetics samples, as well as from cell culture fluid and buffers. The lowest concentrations assayed were 100 ng ml(-1). Quantitation is reproducible, with a coefficient of variation of 2%. The procedure was used to develop a pharmacokinetic profile for the clearance of TNFr-IgG in humans and cynomolgus monkeys. Sufficient material was recovered such that the glycoforms could be identified. Additionally it has been used for process monitoring. The results compared favorably with data generated by ELISA. Optimization of the method and results are presented. PMID- 10379654 TI - Determination of 5-hydroperoxyeicosatetraenoic acid produced in rat basophilic leukemia cell line RBL-2H3 by high-performance liquid chromatography with chemiluminescence detection. AB - A simple and sensitive method, applicable to quantification of 5 hydroperoxyeicosatetraenoic acid (5-HPETE) produced in cells has been developed using high-performance liquid chromatography on a silica gel column with chemiluminescence detection. 5-HPETE was clearly separated from other positional isomers of HPETEs and hydroxyeicosatetraenoic acids with hexane-isopropanol acetic acid (97:3:0.01, v/v) as the mobile phase. The lower limit of detection was about 100 pg. 5-HPETE produced in 10(7) cells of RBL-2H3 cells stimulated with A23187 was determined as 480+/-30 pg. In the present study, 5-HPETE, which occurs naturally, was detected and quantitated for the first time in intact cells. PMID- 10379655 TI - Determination of N-methyl-D-aspartate in tissues of bivalves by high-performance liquid chromatography. AB - The natural occurrence of N-methyl-D-aspartate (NMDA) is limited to the foot muscle of Scapharca broughtonii; it is a well known compound for its neuroexitatory activity. This paper describes a high-performance liquid chromatographic (HPLC) method for the determination of NMDA in biological extracts. The method involves removal of neutral and basic substances by anion exchange chromatography and removal of acidic primary amino acids by treatment with o-phthalaldehyde before derivatization with (+)-1-(9-fluorenyl)ethyl chloroformate, followed by HPLC with isocratic elution with a selected mobile phase that separates the two diastereomers formed. The identity of the detected NMDA has been confirmed by a procedure using (-)-1-(9-fluorenyl)ethyl chloroformate as a derivatizing agent. The identification has been further supported by the disappearance of the peak of the NMDA derivative by pretreatment of the sample with D-aspartate oxidase. Application of the method has shown the presence of NMDA in several tissues of S. broughtonii and Scapharca subcrenata. PMID- 10379656 TI - Fractionation of polyclonal antibodies to fragments of a neuroreceptor using three increasingly chaotropic solvents. AB - We have developed specific antibodies against fragments of anaplastic lymphoma kinase (ALK) in order to develop tools for characterizing the expression and biological function of this orphan receptor. The first fragment consisted of residues 280 to 480 of the murine extracellular domain, was expressed in Escherichia coli (E. coli), purified in the presence of urea from the pellet of mechanically lysed cells and injected into rabbits as an unfolded protein in urea. The second fragment consisted of residues 1519 to 1619 of the murine sequence, corresponding to the C-terminal side of the kinase domain. It was expressed in E. coli as a soluble glutathione-S-transferase fusion protein, purified from the supernatant of broken cells and injected into rabbits as a folded protein. Both antisera were purified using antigen affinity chromatography, with the polyclonal antibodies eluted stepwise using three different buffers, 0.1 M glycine, pH 2.9, followed by 7 M urea, pH 4, followed by 6 M guanidine-HCl (GdnHCl), pH 4. Antisera prepared against either antigen contained antibodies that eluted in each of the three pools, indicating that solvents more chaotropic than acid were required to elute antibody populations that were tightly bound to the antigen column. All three antibody pools were reactive towards their respective antigens upon Western blot analysis. Purified polyclonal antibodies (pAbs) to both fragments also recognized the full-length protein expressed in Chinese hamster ovary cells. In every case, the pAbs eluting in GdnHCl were the most sensitive for detecting full-length ALK. PMID- 10379657 TI - High-performance liquid chromatographic-fluorimetric assay for cathepsin A (lysosomal protective protein) activity. AB - A rapid and sensitive assay for the determination of cathepsin A activity is reported. This method is based on fluorimetric detection of a dansylated peptide, 5-dimethylaminonaphthalene-1-sulfonyl-L-Phe, enzymatically formed from the substrate 5-dimethylaminonaphthalene-1-sulfonyl-L-Phe-L-Leu, after separation by high-performance liquid chromatography using a C18 reversed-phase column and isocratic elution. This method is sensitive enough to measure 5 dimethylaminonaphthalene-1-sulfonyl-L-Phe at concentrations as low as 300 fmol, yields highly reproducible results and requires less than 7.0 min per sample for separation and quantitation. The optimum pH for cathepsin A activity was 4.5-5.0. The Km and Vmax values were respectively 14.9 microM and 27.91 pmol/microg/h with the use of enzyme extract obtained from mouse kidney. Cathepsin A activity was strongly inhibited by Ag+, Hg2+, diisopropylfluorophosphate and p chloromercuriphenylsulphonic acid. Among the organs examined in a mouse, the highest specific activity of the enzyme was found in kidney. The sensitivity and selectivity of this method will aid in efforts to examine the physiological role of this peptidase. PMID- 10379658 TI - Determination of clenbuterol in bovine urine using gas chromatography-mass spectrometry following clean-up on an ion-exchange resin. AB - A gas chromatographic-mass spectrometric method was developed for the determination of residues of clenbuterol in bovine urine. The method involves a simple cation-exchange clean-up and concentration of clenbuterol in the acidified urine, followed by ethyl acetate extraction. The analyte is determined as the di trimethylsilyl derivative and quantitated against an internal standard of penbutolol. Using a 5-ml sample of urine, a detection limit of 0.07 ng/ml can be achieved with recoveries close to 100% for fortification levels of 0.2 and 1 ng/ml. By increasing the sample volume to 50 ml, a detection limit below 0.01 ng/ml was achievable with recovery averaging 70%. The coefficient of variation of the assay ranged from 15% at 0.01 ng/ml (50-ml sample) to 6% at 1 ng/ml (5-ml sample). It was demonstrated that the method can detect the presence of clenbuterol in bovine urine at sub-ppb (ng/ml) levels using low resolution GC-MS with electron impact (EI) ionization. PMID- 10379659 TI - Quantification of residual dimethyl sulfoxide in supernatants of haematopoietic stem cells by capillary zone electrophoresis. AB - Dimethyl sulfoxide (DMSO) is a chemical compound that is used to preserve haematopoietic stem cells during freezing at -180 degrees C. As DMSO is largely removed by washing before reinjection of cells into a patient, accidents (notably cardiovascular) are infrequent. The lack of a method for evaluating the residual quantities of this product led us to develop a technique for assaying DMSO by capillary zone electrophoresis without extraction. This simple, rapid and precise technique was applied to the supernatant of cell pellets of thirteen patients before and after washing. PMID- 10379660 TI - Gas chromatographic-mass spectrometric analysis of perillyl alcohol and metabolites in plasma. AB - Perillyl alcohol (POH), a metabolite of d-limonene and a component of the lavender oil, is currently in Phase I clinical trials both as a chemopreventative and chemotherapeutic agent. In vivo, POH is metabolized to less active perillic acid (PA) and cis- and trans-dihydroperillic acids [DHPA, 4-(1'-methylethenyl) cyclohexane-1-carboxylic acid]. Previous pharmacokinetic studies using a GC-MS method detected POH metabolites but not POH itself; thus these studies lacked information on the parent drug. The present report describes a sensitive GC-MS method for the quantitation of POH and metabolites using stable-isotopically labeled internal standards. The residue obtained from CH2Cl2 extraction of a plasma sample was silylated. The products were separated on a capillary column and analyzed by an ion-trap GC-MS using NH3 chemical ionization. POH-d3 was used as the internal standard for POH while 13C-PA-d2 was used as the internal standards for the metabolites. The quantitation limits for POH, PA, cis- and trans-DPA were <10 ng/ml using 1-2 ml plasma. The assay was validated in rat and human plasma. The assay was linear from 2 to 2000 ng/ml for POH, 10 to 1000 ng/ml for PA and trans-DHPA, and 20 to 1000 ng/ml for cis-DHPA monitored. The within run and between-run coefficients of variation were all <8%. Preliminary pharmacokinetic data from a rat following i.v. administration of POH at 23 mg/kg and from a patient receiving POH at 500 mg/m2 p.o. was also provided. Intact POH, PA, cis- and trans-DHPA were all detected in plasma in both cases. Two new major metabolites were found in human and one in the rat plasma. PMID- 10379661 TI - Simple liquid chromatographic method for the analysis of the blood brain barrier permeability characteristics of ceftriaxone in an experimental rabbit meningitis model. AB - A simple LC method was developed and validated for the analysis of ceftriaxone in aqueous and biological samples. Chromatographic separation was achieved on a reversed-phase C18 microbore column (Hypersil 5 microm, 200x2.1 mm) with UV detection at 270 nm. This isocratic system was operated at ambient temperature and required less than 10 min of chromatographic time. The flow-rate was maintained at 0.5 ml min(-1). Cetyltrimethylammonium bromide (0.01 M) was utilized as the ion-pairing agent. For the analysis of the drug in the aqueous system, the mobile phase consisted of methanol-acetonitrile-phosphate buffer, pH 7.4 (20:20:60, v/v/v). The plasma and CSF systems used the same mobile phase constituents in a slightly different ratio (30:40:30, v/v/v). Lidocaine was used as an internal standard and the peak height ratios of the drug to that of the internal standard were linear over the concentration range of 0.0 to 16 microg ml(-1) only in the case of aqueous systems. Within-day and day-to-day relative standard deviations ranged from 0.3 to 2.2% and 1.1 to 5.9%, respectively. This method was used to: (1) quantify ceftriaxone in an aqueous system, in rabbit plasma using a simple protein precipitation procedure, and in the CSF; (2) evaluate the permeability characteristics of ceftriaxone across the blood-brain barrier through quantification of ceftriaxone in the CSF using a microdialysis sampling technique; and (3) analyze the effects of dexamethasone (a synthetic fluorinated corticosteroid used for the relief of cerebral edema) on the permeability of ceftriaxone across the blood brain barrier through quantification of ceftriaxone in the dexamethasone-treated animals with meningitis. PMID- 10379662 TI - Simultaneous analysis of ketamine and bupivacaine in plasma by high-performance liquid chromatography. AB - A reversed-phase HPLC technique for the simultaneous measurement of both bupivacaine and ketamine in plasma is described. Plasma samples (0.5 ml) were prepared using a rapid and simple back-extraction technique. Resolution of both analytes and the internal standard, desipramine, from medicines coadministered to surgical paediatric patients was obtained using a 5 microm cyano (CN) (250x4.6 mm) column and a mobile phase comprising methanol-acetonitrile-orthophosphoric acid-0.01 M sodium dihydrogenphosphate (200:80:2:718). Good sensitivity for both analytes was observed using UV detection at a wavelength of 215 nm. The method has been validated according to the criteria established by the Journal of Chromatography B. PMID- 10379663 TI - Determination of the geometrical diarylpropenamine isomers in feces by high performance liquid chromatography. AB - Diarylpropenamine derivatives are a class of compounds which have been evaluated as potential drug candidates. Here a specific and reproducible HPLC method for the determination of cis- and trans-isomers of the unsubstituted derivative, 3 (4'-bromo-[1,1'-biphenyl]-4-yl)-3-(4-X-phenyl-N,N-dimethyl-2-propen -1-amine (I, where X=H) in feces is described. The analyte I and internal standard, nitro derivative (II, where X=NO2), were isolated from the basified biological matrix using a liquid-liquid extraction with ethyl acetate followed by a solid-phase procedure performed on a silica cartridge. The organic phase was evaporated to dryness, the residue was reconstituted in mobile phase and injected into the HPLC system. The analytes were eluted with ethyl acetate-hexane-triethylamine (59:40:1) in HPLC column (silica) and detected by UV spectrophotometry at 272 nm. Linearity, precision and accuracy data for feces standards after extraction were acceptable. The method has been applied to analyses of feces samples from rats dosed with I, in which it could be anticipated that fecal excretion is quantitatively the major route for I elimination. PMID- 10379664 TI - Determination of a new oral cephalosporin, S-1090, in human plasma and urine by direct injection high-performance liquid chromatography with ultraviolet detection and column switching. AB - Direct injection high-performance liquid chromatographic (HPLC) methods with column switching and UV detection were developed for the rapid and accurate determination of S-1090 in human plasma and urine. An internal-surface reversed phase pre-column and a C18 analytical column were used for the plasma assay. Two pre-columns packed with cyano and phenyl materials and a C18 analytical column were used for the urine assay. The calibration curves for plasma and urine assays were linear in the ranges 0.09-9 microg/ml and 0.5-100 microg/ml of S-1090, respectively. The relative standard deviations for plasma and urine assays were less than 6% with low relative errors. The established HPLC methods were demonstrated to be useful for clinical pharmacokinetic studies after oral administration of S-1090. PMID- 10379665 TI - High-performance liquid chromatographic analysis of the anti-tumor agent SCH 66336 in cynomolgus monkey plasma and evaluation of its chiral inversion in animals. AB - SCH 66336 is a novel non-cytotoxic anti-tumor agent that is in phase I/II clinical trials for the treatment of solid tumors. This compound is a single enantiomer with one chiral center. Prior to evaluation of this drug candidate in man, it was necessary to evaluate its pharmacokinetics and possible chiral inversion in animals. Thus, high-performance liquid chromatographic (HPLC) methods have been developed for its determination in cynomolgus monkey plasma and for the evaluation of its chiral inversion in rats and cynomolgus monkeys. The achiral HPLC analysis involved extraction with 30% methylene chloride in hexane followed by separation on a CN column and quantitation by UV absorbance at 280 nm. The method was linear over a concentration range of 0.1 to 20 microg/ml in monkey plasma. The chiral HPLC analysis involved the use of a Chiralpak AD column set at 39 degrees C with a mobile phase of hexane-ethanol-diethylamine mixture and a UV detector set at 280 nm. Plasma samples were subjected to solid-phase extraction on a C2 cartridge prior to HPLC analysis. The method was linear over a concentration range of 0.25 to 10 microg/ml in rat and cynomolgus monkey plasma for both enantiomers. Both methods showed good linearity (r2>0.99), accuracy (bias< 13%) and precision (CV<12%). Chiral HPLC analysis indicated that SCH 66336 was not subjected to chiral inversion in rats and cynomolgus monkeys. PMID- 10379666 TI - Determination of iothalamate in rat urine, plasma, and tubular fluid by capillary electrophoresis. AB - A method for the quantitative determination of iothalamate (IOT) in rat urine, plasma and tubular fluid by capillary zone electrophoresis (CE) has been developed and validated. Samples of urine and tubular fluids were diluted with water and samples of plasma were deproteinized with two volumes of acetonitrile containing the internal standard, p-aminobenzoic acid (PABA). A BioFocus 2000 system (Bio-Rad, Hercules, CA, USA) was used. The UV detector was set at 254 nm. The samples were loaded into uncoated fused-silica capillary (40 cmx50 microm) by pressure injection. A borate buffer [20 mM, pH 12 (pH adjusted with 1.0 M NaOH)] was used as the electrophoretic buffer. The typical analytical conditions were: voltage, 22 kV; injection, 9 psixs; capillary and carousel temperatures were 20 degrees C and 18 degrees C respectively. The linear relationship was observed between time-corrected peak area of IOT in water and urine or the corrected peak area ratio of IOT to PABA in plasma and the nominal concentration of IOT with correlation coefficient greater than 0.999. The intra- and inter-day coefficients of variation (CV) were less than 8%. The concentration of IOT in plasma, urine and tubular fluid determined by CE can be used for estimation of whole kidney and single nephron clearances. PMID- 10379667 TI - High-performance liquid chromatography separation and quantitation of ofloxacin enantiomers in rat microsomes. AB - A sensitive, simple and accurate method for determination of enantiomers of ofloxacin in microsomal incubates was developed by chiral ligand-exchange RP-HPLC with fluorescence detection to examine stereoselective metabolism of ofloxacin in the glucuronidation process. The C18 stationary phase was used as analytical column. The solution of chiral mobile phase additive was made up of 6 mM L phenylalamine mixed with 3 mM CuSO4 in water. Mobile phase consisted of the solution of chiral mobile phase additive-methanol (86:14). The fluorescence detector was operated at lambda(ex) 330 nm and lambda(em) 505 nm. The flow-rate of mobile phase was set at 1.0 ml/min. The achiral ODS column offers good separation of the two enantiomers in less than 25 min. The recovery of the assay was 97.9+/-6.1% (n=10) for S-ofloxacin and 99.6+/-6.0% (n = 10) for R-ofloxacin. The method provides a high sensitivity and good precision (RSD<10%). The LOD was 0.6 microM for both enantiomers and the LOQ was 5.70+/-0.45 microM (n=8) for S ofloxacin and 5.66+/-0.47 microM (n=8) for R-ofloxacin. The standard curves showed excellent linearity over the concentration range 5.5-2078 microM for S-(-) ofloxacin and R-(+)-ofloxacin. The enantioselective method developed has been applied to determine the stereoselectivity of glucuronidation metabolism of ofloxacin optical isomers in rat liver microsomes. PMID- 10379668 TI - Secretion of interferon-tau by bovine embryos in long-term culture: comparison of in vivo derived, in vitro produced, nuclear transfer and demi-embryos. AB - Interferon-tau (IFNtau) is the pregnancy recognition signal of bovine embryos, inhibiting luteolysis. We studied trophoblastic growth and IFNtau secretion of embryos with different developmental potential, i.e., in vivo derived and in vitro produced embryos, cloned embryos and demi-embryos, to evaluate if the ability of secreting IFNtau might be responsible for differences in pregnancy rates after transfer of these categories of embryos to recipients. Day 8 embryos of excellent quality were individually placed in microdrops of buffalo rat liver cell-conditioned medium and maintained for up to 23 days. Embryos were observed on Days 11, 15, 19 and 23, the mean diameter (2r) of attached and spherical embryos was measured, and their trophoblastic area was calculated as r2pi or 4r2pi, respectively. Simultaneously, medium was changed and the IFNtau levels of conditioned media were determined using a bioassay of antiviral activity. Trophoblastic area was smaller (P < 0.05) in demi-embryos than in all other groups, which exhibited similar trophoblastic growth until Day 19. However, on Day 23 trophoblastic area of in vivo derived embryos was more than twice (P < 0.05) as large as those of in vitro produced and nuclear transfer (NT) embryos. IFNtau levels increased only slowly with time in culture of demi-embryos. By contrast, the level of IFNtau doubled from Day 11 to Day 15 in conditioned media from all other groups of embryos. The linear increase in IFNtau production of vivo and in vitro derived embryos continued until the end of the culture period, whereas conditioned media from NT embryos contained significantly (P < 0.05) less IFNtau activity on Days 19 and 23 than those of the former two groups. Our results demonstrate different capabilities of secreting IFNtau for in vivo derived and in vitro produced embryos vs. NT and demi-embryos, which may--at least part--be responsible for the differences in pregnancy rates after transfer to recipients. PMID- 10379669 TI - Measurements of bovine sperm velocities under true anaerobic and aerobic conditions. AB - Velocities of bovine spermatozoa in a medium containing glucose were similar under true anaerobic and aerobic conditions. Spermatozoa were not able to sustain motility under anaerobic conditions when glycolysis was inhibited, but regained motility when re-aerated. This demonstrates that immobilisation was due to lack of oxygen and that conditions under which motility was analysed were truly anaerobic. Sperm motility parameters were not significantly different in the presence and absence of 4 microM antimycin A and 4 microM rotenone when glucose was present in the medium. After each incubation, functionality of sperm mitochondria was assayed by washing sperm into the medium which supported respiration but not glycolysis, and motility was visually assessed. All sperm samples were highly motile in this medium indicating that their mitochondria were functional. When glycolysis was inhibited, antimycin and rotenone abolished sperm motility immediately after addition. Bovine sperm can maintain similar levels of motility aerobically and anaerobically if a glycolysable substrate is available. Available data on bovine sperm energetics support this view. PMID- 10379670 TI - Superovulation induction in Japanese Black cattle by a single intramuscular injection of hMG or FSH dissolved in polyvinylpyrrolidone. AB - The efficacy of a single intramuscular dose of 450 or 600 international units (IU) of human menopausal gonadotropin (hMG) or 30 mg of follicle stimulating hormone (FSH), each dissolved in 30% polyvinylpyrrolidone K-30 (PVP), for superovulation treatment was compared to that of superovulation induction by administration of a total dose of 600 IU hMG given in declining doses twice daily over a 3-day period. A total of 48 Japanese Black cows were used for the investigation. Oestrus was observed within 60 h after PGF2alpha administration in all cows in the hMG groups. In the hMG group that received a single dose of 600 IU hMG (n = 12), oestrus was observed less than 36 h after treatment in one cow. In contrast, oestrus was not observed in 3 of the 12 cows (25%) in the FSH group. Neither the average number of recovered ova/embryos nor the number of transferable embryos per collection differed significantly among the hMG groups. However, the average number of transferable embryos was not significantly higher in cows treated with a single dose of 600 IU of hMG than in cows treated with a single 30 mg dose of FSH (7.5+/-4.5 vs. 2.1+/-2.8). The number of cows from which more than three excellent grade embryos were collected was highest in the group that received a single dose of 600 IU hMG (9/12, 75%) and lowest in the group that received a single 30 mg dose of FSH (2/9, 22%). The differences between groups in the percentages of cows with three or more excellent embryos between treatments were not statistically significant. The proportion of recovered ova/embryos classified as excellent was highest in the group that received 600 IU hMG in declining doses and lowest in the group that received a single 30 mg dose of FSH (55.2% vs. 30.2%; P < 0.05). The recovery rate of unfertilized ova was lowest in the group that received a single dose of 600 IU hMG and highest in the group received a single 30 mg dose of FSH (18.3% vs. 48.8%; P < 0.05). Although the differences in recovery results between the groups were not statistically significant, the recovery rates in hMG groups were higher than that the FSH group. These findings suggest that superovulation can be induced adequately in Japanese Black cows using one injection of 450 to 600 IU hMG dissolved in PVP. PMID- 10379672 TI - Effect of adrenocorticotrophic hormone (ACTH1-24) on ovine pituitary gland responsiveness to exogenous pulsatile GnRH and oestradiol-induced LH release in vivo. AB - The present experiment was designed to determine if and how exogenous ACTH replicates the effects of stressors to delay the preovulatory LH surge in sheep. Twenty-four hours after oestrous synchronisation with prostaglandin in the breeding season, groups of 8-9 intact ewes were injected with 50 microg oestradiol benzoate (0 h) followed 8 h later by 3 injections of saline or GnRH (500 ng each, i.v.) at 2 h intervals (controls). Two further groups received an additional 'late' injection of ACTH (0.8 mg i.m.) 7.5 h after oestradiol, i.e., 0.5 h before the first saline or GnRH challenge. To examine if the duration of prior exposure to ACTH was important, another group of ewes was given ACTH 'early', i.e. 2.5 h before the first GnRH injection. The first GnRH injection produced a maximum LH response of 1.9+/-0.4 ng/ml which was significantly (p < 0.01) enhanced after the second and third GnRH challenge (7.1+/-1.5 ng/ml and 7.0+/-1.7 ng/ml, respectively; 'self-priming'). Late ACTH did not affect the LH response after the first GnRH challenge (1.9+/-0.4 vs. 1.8+/-0.3 ng/ml; p > 0.05) but decreased maximum LH concentrations after the second GnRH to 35% (7.1+/-1.5 vs. 4.6+/-1.1 ng/ml; p = 0.07) and to 40% after the third GnRH (7.0+/-1.7 vs. 4.0+/-0.8 ng/ml; p = 0.05). When ACTH was given early, 4.5 h before the second GnRH, there was no effect on this LH response suggesting that the effect decreases with time after ACTH administration. Concerning the oestradiol-induced LH surge, exogenous GnRH alone delayed the onset time (20.5+/-2.0 vs. 27.8+/-2.1 h; p > 0.05) and reduced the duration of the surge (8.5+/-0.9 vs. 6.7+/-0.6 h; p > 0.05). The onset of the LH surge was observed within 40 h after oestradiol on 29 out of 34 occasions in the saline +/- GnRH treated ewes compared to 11 out of 34 occasions (p < 0.05) when ACTH was also given, either late or early. In those ewes that did not have an LH surge by the end of sampling, plasma progesterone concentrations during the following oestrous cycle increased 2 days later suggesting a delay, not a complete blockade of the LH surge. In conclusion, we have revealed for the first time that ACTH reduces the GnRH self-priming effect in vivo and delays the LH surge, at least partially by direct effects at the pituitary gland. PMID- 10379673 TI - Foetal loss in dairy goats: function of the adrenal glands, corpus luteum and the foetal-placental unit. AB - To investigate the causes and mechanisms of foetal loss in Norwegian dairy goats, blood parameters in 40 goats that lost foetuses were compared with those in 40 goats that experienced a normal pregnancy. High mean levels of 15-ketodihydro PGF2alpha, and low mean levels of oestrone sulphate throughout pregnancy were associated with foetal loss. The mean oestrone sulphate level was low before abortion, and the distinct peak that occurred at parturition in the control goats was not observed in connection with abortion. Association of other blood parameters with foetal loss was not detected. Infectious agents and toxins did not appear to be major causes of foetal loss in this study. The normal level of progesterone and cortisol in goats with foetal loss indicated that the function of the corpus luteum and adrenal glands, respectively, were not disturbed. The rapid decline in progesterone level associated with foetal loss may therefore be a result, rather than the cause, of foetal death. The lowered level of oestrone sulphate and elevated level of 15-ketodihydro-PGF2alpha in goats with foetal loss clearly indicated that the endocrine foetal-placental function was disturbed. PMID- 10379671 TI - Development of a sensitive enzymeimmunoassay (EIA) for FSH determination in bovine plasma. AB - A highly sensitive enzymeimmunoassay (EIA) procedure for FSH determination in bovine plasma on microtiterplates using the biotin-streptavidin amplification system and the second antibody coating was developed. Biotin was coupled to FSH and used to bridge between streptavidin-peroxidase and the immobilized antiserum in the competitive assay. The EIA was carried out directly in 50 microl of bovine plasma and compared with an established radioimmunoassay (RIA) employing 100 microl plasma. Same FSH standards and FSH specific antiserum were used in both procedures. FSH standards prepared in hormone free plasma were used. The sensitivity of the EIA procedure was 6.25 pg/well FSH which corresponded to 125 pg/ml plasma; the 50% relative binding sensitivity was seen at 200 pg/well. In comparison to RIA, the EIA was at least four times more sensitive besides requiring 6 times less FSH specific antiserum. Plasma volumes for the EIA ranging from 12.5 to 50 microl did not influence the shape of the standard curve even though a slight drop in the OD450 was seen with higher plasma volumes. When both EIA and RIA methods were used to measure FSH in cows, the levels were detectable only by the EIA procedure. The assay detects high and low plasma FSH levels within the physiological variation as well as changes in plasma FSH after stimulation with a GnRH analog. In conclusion, in addition to being non radioactive and low cost in nature, the method offers several advantages over the conventional FSH RIA procedure; these are (a) higher sensitivity, (b) less labour and time saving, (c) more economical use of precious FSH antiserum and (d) long shelf-life of the biotinyl-FSH label (in contrast to the short half life of iodinated FSH in RIA). PMID- 10379674 TI - Seasonal effects on fertility and ovarian follicular growth and maturation in camels (Camelus dromedarius). AB - Camels are said to be seasonal breeders, but the extent to which season interferes with food supply to affect ovarian function is not fully documented. Hence, the three aims of this study were: (1) to define the breeding season of camels maintained in semi-arid conditions in southern Morocco; (2) to relate the proportion of females with active ovaries (i.e., with follicles > 5 mm), with ovulatory (11-17 mm) or cystic (> 18 mm) follicles to age and body conditions score; (3) to study the consequences of the interactions between age and body conditions score on the proportion of females ovulating and conceiving; and (4) to compare follicular maturation, using in vitro steroidogenesis by intact follicles as a marker during the transition into the breeding season (October) and peak breeding season (March). There was a clear breeding season in the two flocks studied, since over 80-90% of the matings occurred during the period from mid-November to mid-April. Collection of ovaries at slaughter (n = 238) demonstrated a significant seasonal effect on the proportion of females with active ovaries (increasing from 73.5% in October-December to 89% in January-May), but no changes in the proportion of females with ovulatory follicles. Lean females (BCS < 2.5) had a delayed initiation of ovarian function in October December. In addition, the proportion of females with cystic follicles was also affected by season (peaking during April-May). Neither age nor body condition modulated the frequency of cysts. Finally, the proportion of females conceiving increased steadily as season progressed (peaking at 57% in April-May). Body condition score did not affect this proportion, but young females (< or = 5 years old) had a low ability to conceive. Morphological features of large follicles were unaffected by season. Ovulatory follicles contained around 10(7) granulosa and theca cells. In vitro testosterone output by intact follicles was unrelated to follicle size and season. In vitro oestradiol output increased with increasing follicle size and was larger in follicles obtained during peak breeding season than at its initiation. This may indicate that early breeding season follicles display a low aromatase activity in their granulosa cells. Whether the low oestradiol output of early breeding season follicles is resulting in the low fertility observed at this period remains to be determined. PMID- 10379675 TI - Reproductive performance of farmed red deer (Cervus elaphus) in New Zealand. IV. Biological markers as risk factors for yearling and adult hind conception. AB - A 2-year observational study of 15 red deer (Cervus elephus) farms was carried out in New Zealand from March 1992. In each year of study, approximately 1650 hinds were individually monitored for reproductive success. During farm visits in March 1992 and 1993, five yearling and five adult hinds per farm were randomly selected and blood sampled to define their haematological, biochemical and blood mineral profile. Faecal samples were taken for parasite egg and larvae count. Biological markers potentially affecting the probability of conception before May 1 or of conception that year were investigated separately using multivariable logistic regression analysis. Adult hinds with low serum phosphorus concentrations were more likely to conceive before May 1. Lower conception rates were observed in yearling hinds when blood glutathione peroxidase, serum vitamin B12, and serum albumin concentrations were low, and when faecal lungworm larval counts were high. While these associations have yet to be proven as causal, data suggests that monitoring and maintaining adequate blood elements, and controlling internal parasites in yearling hinds, may assist farmers to achieve optimum reproductive performance in farmed red deer herds. PMID- 10379676 TI - Effects of oestrogen supplementation and space restriction on PGF2alpha-induced nest-building in pseudopregnant gilts. AB - This study examined the role of oestrogen supplementation on PGF2alpha-induced nest-building in pseudopregnant gilts. Oestradiol valerate (5 mg/day) injections were given on Days 11-15 of the oestrous cycle to induce pseudopregnancy. A further series of injections of either oestradiol valerate (5 mg/day) or vehicle were given on days 44-46 of pseudopregnancy to reflect more closely the hormone profile seen in pregnancy. Nest-building was induced by a single intramuscular injection of 15 mg of PGF2alpha (Lutalyse) on Day 47 of pseudopregnancy. The gilts were housed in pens (2.8 x 1.7 m) containing straw in experiment 1 or chronically confined in crates (0.6 x 1.7 m) that did not contain straw on days 44-48 of pseudopregnancy for experiment 2. Oestrogen supplemented gilts had significantly higher concentrations of circulating 17beta-oestradiol on day 47 of pseudopregnancy but there were no significant differences between treatments for circulating levels of prolactin, progesterone, cortisol or oxytocin, or for any behavioural measure in either experiment. These results indicate that there is no direct effect of supplementing already pseudopregnant gilts with oestradiol valerate on PGF2alpha-induced nest-building. The results also show that the pre partum environment has a pronounced effect on nest-building behaviours and that non-pregnant pigs might be a useful model for pre-partum nest-building in this species. PMID- 10379677 TI - Characterization of the semen quality of postpuberal boars with spontaneous unilateral abdominal cryptorchidism on the right side. AB - In recent studies, we found that the ectopic testis from postpuberal boars with unilateral abdominal cryptorchidism does not produce sperm. Therefore, in these males, the seminal characteristics can be used as indicators of the activity of the scrotal testis and its epididymis and also the accessory glands. The semen quality (ejaculate volume, cell-rich fraction volume, sperm concentration, sperm vitality, sperm motility, sperm morphology and cephalic stability of spermatozoa) was evaluated in healthy postpuberal boars and in postpuberal boars with unilateral abdominal cryptorchidism on the right side. In comparison with the healthy boars, the unilateral abdominal cryptorchid boars showed a significant decrease of the ejaculate volume, sperm concentration and sperm motility. The low sperm concentration indicated that unilateral abdominal cryptorchidism severely impairs the sperm production of the scrotal testis. The decrease of ejaculate volume was attributed to an abnormal activity of the accessory glands. The alterations in sperm motility develop as a result of dysfunctions in the epididymal epithelium and/or the accessory glands. The sperm vitality, sperm morphology and cephalic stability of spermatozoa maintained normal values; therefore, at testicular level, despite the low sperm production, the germ cell differentiation is not disturbed. At epididymal level, the morphological maturation of spermatozoa is not altered. PMID- 10379678 TI - The hypoosmotic swelling test performed with coulter counter: a method to assay functional integrity of sperm membrane in rainbow trout. AB - The hypoosmotic swelling test (HOS) is one of the methods used to evaluate sperm quality in mammals. This test is based on the swelling ability that functional spermatozoa have when submitted to hypoosmotic solutions. Only a slight increase in size is caused in rainbow trout spermatozoa in such conditions and it is not possible to distinguish between reactive cells (cells who were capable to increase in volume) and non-reactive cells (did not increase in volume) under light microscopy. In our approach we have used the coulter counter to verify the effectiveness of the HOS test in this species. Semen was diluted in different hypoosmotic solutions (50, 100, 150, 200, 250 and 320 mosM/kg) and cell volume measured at different times after dilution (30 s, 2, 5, 10, 20, and 30 min). The higher percentage of reactive cells was achieved with the 100 mosM/kg solution and swelling occurred before 30 s. Even with this solution, the small increase in cell size caused the overlapping of volumes from swollen and non-swollen spermatozoa. In order to analyse the data and to choose a parameter suitable for assessing cell reactivity, the test was performed in samples containing known rates of live/dead cells. Two parameters were analysed after swelling: the increase in volume and the percentage of cells over a standard volume (reactive cells). Results showed a high correlation between the percentages of reactive cells and the known rate of live cells (r2 = 0.65). This fact suggests that HOS test could be used to analyse the integrity and functionality of rainbow trout fresh sperm. To study the reliability of this test in cryopreserved sperm, simple linear regressions were made between cell viability determined by Hoechst 33285 dye and the two parameters obtained from coulter counter data. No significant correlation was observed in either case, showing that structural and functional integrity do not correlate after freeze/thaw. Consistently, the HOS test is not a reliable method to evaluate cryopreserved sperm quality in rainbow trout. PMID- 10379679 TI - Effects of lactation, energetic deficit and remating interval on reproductive performance of primiparous rabbit does. AB - Female rabbits (n = 151) were assigned at their first parturition (day 0) to one of three treatment groups (A: non-lactating does and ad libitum feeding, R: non lactating does and restricted feeding, L: lactating does and ad libitum feeding). Additional females (n = 18) were slaughtered at parturition. Experimental females were presented to the male on days 3 (A3, L3 and R3 groups, n = 25, 26 and 26, respectively) or 10 (A10, L10 and R10 groups, n = 25, 25 and 24, respectively) and were slaughtered on day 28 after parturition. Compared to does slaughtered on day 0, adipose tissues were lighter in L (-66%) and R (-32%) females while they were heavier in A females (+40%; P < 0.01). These results suggest that both L and R females were in energy deficit. Receptivity (80% vs. 98%) and conception rate (51% vs. 83%) were lower in L compared to A females (P < 0.01) regardless the day of male presentation. Ovulation rate (-14%) and conception rate (-26%) were lower in R3 than in A3 does (P < 0.05). Ovulation rate was 24% lower in the L10 than in the A10 group (P < 0.01). The uterine contents were lighter (-25%) in L and R than in A does (P < 0.001) regardless the day of male presentation. These results suggest that the energy deficit associated with milk production can partly explain the negative influence of lactation on reproductive performance. PMID- 10379680 TI - Origins and morphogenesis of colorectal neoplasms. AB - Gastrointestinal stem cells are considered pivotal in colonic carcinogenesis. There is evidence to suggest that early microadenomas in the colon are polyclonal in origin. Adenomas, once initiated, enlarge by the process of crypt fission. It is also the main mechanism by which neoplastic clones spread through the colorectal epithelium. Both concepts are important for our understanding of the early events in colonic carcinogenesis. PMID- 10379681 TI - Carrier rate of resistant enterococci in a tertiary care hospital in Norway. AB - The prevalence of resistant enterococci varies geographically. In the present study we looked at the carrier rate of resistant enterococci in the hematology and gastrointestinal surgery units of a tertiary care hospital in Norway. Anal swabs were taken from all 82 hospitalized patients on 4 different dates, at least 4 weeks apart, in 1995. 51% had positive cultures for enterococci. 6% of all patients carried enterococci resistant to ampicillin. 7% carried enterococci with high-level gentamicin resistance. Two strains resistant to vancomycin were found, including the first vanA Enterococcus faecium isolated in a Norwegian hospital. There was a correlation between use of antibiotics and being a carrier of enterococci per se, but the correlation with resistant enterococci did not reach statistical significance owing to the small number of isolates. The carrier rates both for presence of enterococci and for resistant enterococci were generally lower than those found in other studies. PMID- 10379682 TI - A case of endometrioid carcinoma of the fallopian tube mimicking an adnexal tumor of probable Wolffian origin. AB - We report a very uncommon case of endometrioid adenocarcinoma of the fallopian tube that mimicked, based on histology, a female adnexal tumor of probable Wolffian origin (FATPWO). We present our microscopic and immunohistochemical findings, and a review of the literature concerning these two entities. The differential diagnosis can be of great consequence, owing to the very different prognoses of the two tumors, and is based mainly on macroscopic appearance and immunohistochemical profile: epithelial membrane antigen (EMA) and CA125, generally lacking in FATPWO, are expressed in endometrioid adenocarcinoma, thus indicating the mullerian origin of this tumor. PMID- 10379684 TI - An in vitro study of the susceptibility of mobile and cystic forms of Borrelia burgdorferi to metronidazole. AB - The aim of this study was to examine the susceptibility of mobile and cystic forms of Borrelia burgdorferi to metronidazole. Because B. burgdorferi is a microaerobic bacterium like Helicobacter pylori, metronidazole (MZ) was chosen in the susceptibility test. For both microaerobic and aerobic incubation the normal mobile spirochetes were resistant to this antibiotic with an MBC > or = 512 microg/ml. Conversion of mobile spirochetes to cystic forms was not observed when they were incubated with MZ. When they were incubated under microaerobic conditions, the biologically active cystic forms had an MBC > or = 4 microg/ml, but the MBC was > or = 32 microg/ml with aerobic incubation at 37 degrees C. Staining with acridine orange (AO), dark field microscopy (DFM), and transmission electron microscopy (TEM) revealed that the contents of the cysts were degraded when the concentration of MZ was > or = MBC. Some cysts were also ruptured. When incubated with a sufficient concentration of MZ, core structures did not develop inside the cysts, and AO revealed less RNA in the cysts. Our observations may help efforts to treat resistant infections caused by B. burgdorferi with a combination of MZ and other antibiotics in order to eradicate both cystic and mobile forms of B. burgdorferi. PMID- 10379683 TI - Reactions of N-N and N-O compounds with horseradish peroxidase and peroxidases from Mycobacterium tuberculosis. AB - Several N-N-and N-O-containing compounds were analysed for their ability to act as substrates for horseradish peroxidase and peroxidases in Mycobacterium tuberculosis extracts. Aminoguanidine, diaminoguanidine, isoniazid, hydroxylamine and hydrazine were found to be weak substrates for horseradish peroxidase in reaction I and to inhibit the reaction of horseradish peroxidase with hydrogen peroxide. The same compounds inhibited the reaction of Mycobacterium tuberculosis peroxidase-catalase with hydrogen peroxide, and hydroxylamine was found to be a weak substrate for this enzyme. In growth inhibition experiments, diaminoguanidine inhibited the growth of M. tuberculosis H37Rv at 50 microg/mL, but not the growth of two isoniazid-resistant strains. Isonicotinic acid hydroxamate inhibited the reaction of the peroxidases with hydrogen peroxide, but was not itself a substrate and had no growth-inhibitory effects. On the basis of these results we suggest that the effect of isoniazid on growth of M. tuberculosis results from increased oxidative stress due to inhibition of catalase-peroxidase as well as from generation of toxic radicals with the structure [structure in text]. PMID- 10379686 TI - Development of resistance and cross-resistance in Pseudomonas aeruginosa exposed to subinhibitory antibiotic concentrations. AB - The purpose of this study was to compare resistance and cross-resistance development in Pseudomonas aeruginosa isolates from cystic fibrosis (CF) patients to commonly used antipseudomonal antibiotics. Isolates were repeatedly exposed to subinhibitory concentrations of either azlocillin, tobramycin, ceftazidime or ciprofloxacin. On 10 consecutive occasions, samples were removed from the half MIC well of a microtitre plate and regrown in drug-free medium to provide the next inoculum for MIC determination. The increase in MIC at the end of the treatment period was significant (p<0.05) for all selecting antibiotics. Cross resistance to unrelated antibiotics was not observed, but was significant (p<0.05) in all beta-lactams (ticarcillin, piperacillin, ceftazidime and cefsulodin) studied where azlocillin was the selecting antibiotic. The addition of clavulanic acid to ticarcillin and of tazobactam to piperacillin had no effect on cross-resistance. The development of resistance to azlocillin was associated with increased beta-lactamase activity and a change in isoelectric point of the beta-lactamases. The result of this study supports a rotational policy for antipseudomonal antibiotics in CF patients. PMID- 10379685 TI - Molecular detection of EWS-FLI1 chimeric transcripts in Ewing family tumors by nested reverse transcription-polymerase chain reaction: application to archival paraffin-embedded tumor tissues. AB - Chromosomal translocations generating unique chimeric genes are highly characteristic of specific sarcomas, and their use as diagnostic markers has been suggested. From a diagnostic pathologic point of view, detection of such cytogenetic or molecular aberrations applicable to routinely processed archival tissue specimens is considered a powerful tool for tumor diagnosis. To assess the feasibility and reliability of the molecular detection of the transcript originating from the chimeric gene in paraffin-embedded tumor specimens, we performed a nested reverse transcription-polymerase chain reaction (RT-PCR)-based assay to detect the EWS-FLI1 chimeric message in a series of Ewing family tumors. Of 24 paraffin-embedded tumor specimens from 23 cases analyzed, the chimeric message was detectable in 20 (83%) specimens from 20 cases (87%) by this nested RT-PCR assay, whereas none of 7 small round cell tumors not from this family (3 alveolar rhabdomyosarcomas, 2 neuroblastomas, 2 malignant lymphomas) showed detectable chimeric messages. In the sequence analysis of the PCR products, the amplified chimeric messages contained the junctions between exon 7 of the EWS gene and any one of exons 5, 6 and 8 of the FLI1 gene. The detection process was usually completed within 3 days, except for the subseqent sequence analysis. Our results endorse the use of this molecular assay as an ancillary technique in the diagnosis of Ewing family tumors using paraffin-embedded material. PMID- 10379687 TI - C-reactive protein (CRP) response patterns in neonatal septicaemia. AB - C-reactive protein (CRP) is an unreliable diagnostic tool in the early diagnosis of neonatal septicaemia. However, serial measurements have been shown to be useful in monitoring the effectiveness of treatment. The aim of the present study was to investigate whether a specific CRP response pattern to different groups of pathogens could be identified during treatment of neonatal septicaemia. Serial CRP measurements from day 1 to 4 in monomicrobial blood culture-proven episodes of septicaemia were reviewed. In 4416 admissions, 180 out of 206 positive blood cultures were monomicrobial; 121 monomicrobial septic episodes were eligible for final analysis of the CRP response during treatment. A low median (M) value (day 1 to 4) was identified in coagulase-negative staphylococci (CONS) (M=23 mg/l), contrasting with high median values in Staphylococcus aureus (M=58 mg/l), group B streptococci (M=51 mg/l), Escherichia coli (M=51 mg/l) and Candida species (M=76 mg/l) (p<0.001). Median CRP values in the two groups were different for each of the treatment days 1 to 4 (p<0.001). An increase (p<0.001) in CRP during the 24 h before initiation of treatment was a sign of late-onset CONS septicaemia. In episodes where antimicrobial treatment failed, CRP levels were moderately elevated the day prior to treatment start and increased continuously thereafter, whereas successful treatment was generally accompanied by a decline in CRP in less than 4 days. The CRP response to CONS is significantly less pronounced than to other commonly encountered pathogens in neonatal septicaemia. A rise in CRP beyond the third day of empirical treatment should give rise to a suspicion of fungal infection or ineffective antibacterial treatment. PMID- 10379688 TI - Detection of Pneumococcus from whole blood, buffy coat and serum samples by PCR during bacteremia in mice. AB - Whole blood, purified leukocyte fraction and serum were investigated as specimens for the detection of pneumococcal bacteremia by polymerase chain reaction (PCR) in mice. The PCR findings were compared to the blood culture results. Samples were taken from animals 3 and 12 h after intraperitoneal bacterial challenge. The pneumococcal culture was positive in 27% and 77% of blood samples at 3 and 12 h after challenge, respectively. All whole blood samples were positive by PCR at both time points. Of the buffy coat samples, two of the three pools were PCR positive at 3 h and all pools at 12 h after bacterial challenge. In the serum sample group, only 40% of the sera were PCR-positive at 3 h, while at 12 h 90% of the samples were PCR-positive. According to these results, whole blood seems to be the best specimen for the detection of pneumococcal DNA by PCR in bacteremic mice. PMID- 10379689 TI - Loss of bactericidal capacity of long-chain quaternary ammonium compounds with protein at lowered temperature. AB - Amphiphilic betaine esters are quaternary ammonium compounds (QACs) with rapid microbicidal effect, which spontaneously hydrolyze into nontoxic products. thus being referred to as soft antimicrobial agents. The bactericidal effect of 1 decyl (B10), 1-dodecyl (B12), and 1-tetradecyl (B14) betaine esters on Salmonella typhimurium was strongly influenced by temperature, pH and length of hydrocarbon chain. At pH 6.0, presence of 1.5 mM (10% w/v) BSA raised the concentration of B14 for 99% killing (BC2) from 0.006 mM to 1.8 mM. There was a stoichiometric relationship between concentration of BSA and BC2 of B14, indicating that one molecule of B14 was bound per BSA molecule when 99% killing was achieved. When the temperature was lowered to 0 degrees C only minor killing was seen in 1.5 mM BSA at the highest concentration of B14 tested, 57 mM. With B10 at 30 degrees C and pH 6.0, the presence of 1.5 mM BSA raised the bactericidal concentration (BC2) from 0.69 mM to 4.1 mM, and at 0 degrees C and 1.5 mM BSA the BC2 was 11 mM. Thus, the impairment caused of the bactericidal effect of B10 by BSA and lower temperature was less than for B14, since B14 is much more active than B10 at 30 degrees C in the absence of BSA, somewhat more active than B10 at 30 degrees C in the presence of 1.5 mM BSA, and much less active than B10 at 0 degrees C in the presence of BSA. B12 showed properties intermediate between B10 and B14. Lowered pH reduced the bactericidal effect particularly when reduced from pH 5.0 to 4.0 with B10. In the presence of 1.5 mM BSA, the bactericidal effect of 1-dodecyl (DTAB) and 1-hexadecyl (CTAB) trimethylammonium bromide decreased in the same manner as for B10 and B14, respectively. Increasing the time of incubation at 0 degrees C to 50 min, a 99% killing effect was seen with 17 mM CTAB, whereas the same killing effect was reached in 8 min with 17 mM DTAB. Binding of [3H]CTAB to S. typhimurium was also reduced at 0 degrees C in the presence of BSA. Thus, in the presence of 1.5 mM BSA, QACs with the longer hydrocarbon chain were most efficient at 30 degrees C, whereas at 0 degrees C those with the shorter hydrocarbon chain were most active. Consequently, QACs with shorter tails should be used for disinfection in the presence of proteins at lower temperatures. PMID- 10379690 TI - Nonparametric step-down test procedures for finding minimum effective dose. AB - Nonparametric versions of normal theory step-down multiple-test procedures for inferring minimum effective dose (see Tamhane et al. (1)) were developed and studied by Monte Carlo simulation. Two types of step-down testing procedures were examined. For both procedures, pairwise, linear, or Helmert contrasts of mean ranks were studied. All nonparametric step-down procedures controlled familywise error rate under normal, double exponential, and exponential distributions. Specific recommendations based on power and bias comparisons of nonparametric methods among themselves, with Shirley's (2) test, and with parametric step-down multiple-test procedures are given. PMID- 10379691 TI - Estimating sample sizes for continuous, binary, and ordinal outcomes in paired comparisons: practical hints. AB - Paired data occur in crossover trials and matched case-control studies, and it is rare to find studies reporting sample size calculations associated with these types of studies, despite recommendations from editors that sample size calculations should be justified. In this article we describe some simple formulas and strategies for calculating the number of patients that should be entered into a matched or paired study when the outcome measures are continuous, binary, or ordinal. PMID- 10379692 TI - Pharmacodynamic analysis of analgesic clinical trials: nonlinear mixed-effects logistic models. AB - In the development of an analgesic product, placebo-controlled clinical trials in patients with defined pain are used to study the dose-time-response relationship of the drug. In such trials, the response is usually an ordered categorical variable with longitudinal and subject-specific repeated measurements. The primary causal variables are time and analgesic concentration. The response may be informative right-hand censored because remedication with a known analgesic may be given if a patient has inadequate pain relief. Mixed-effects logistic models are used to estimate the probabilities of having certain pain relief or pain severity scores. A jackknife method is proposed to estimate the standard errors of parameter estimates. Posterior estimates of these probabilities, or of the scores themselves, allow the evaluation of efficacy for an analgesic. In this evaluation, therapeutic as well as statistical significance is assessed. Two case studies, one focusing on pain relief and the other on pain severity, are used to demonstrate the approach. The level of baseline pain appears to be a determinant of the pattern of response. PMID- 10379693 TI - Nonlinear models for in vitro kill kinetics of antibiotics. AB - In this study, we developed nonlinear regression models to analyze the data generated from an in vitro continuous culture system to assess the kinetics of metronidazole and trovafloxacin in inhibiting the growth of Bacteroides fragilis. The model includes parameters describing the initial shock effect of an antibiotic pulse, the overall antibiotic wash-out rate from the system, and the long-term toxicity of the antibiotic in the environment after one pulse and before the next pulse. PMID- 10379694 TI - Effect of investigator bias on the power and level of the two-sample Z-test. AB - The use of subjective measures in the evaluation of treatment efficacy in clinical trials may introduce bias into the response variables of interest when investigators attempt to guess the treatment that patients are receiving. The bias may be introduced even in the setting of double-blind clinical trials due to the presence of characteristic side effects. The introduction of bias leads to an increase in the power of the statistical test. However, this increase in power is achieved at a considerable increase in the actual level of the statistical test. PMID- 10379696 TI - Nonparametric analysis of covariance for hypothesis testing with logrank and Wilcoxon scores and survival-rate estimation in a randomized clinical trial. AB - Many clinical trials have time-to-event variables as principal response criteria. When adjustment for covariates is of some importance, the relative role of methods for such analysis may be of some concern. For the Wilcoxon and logrank tests, there is an issue of how covariance adjustment can be nonparametric in the sense of not involving any further assumptions beyond those of the logrank and Wilcoxon test. Also of particular interest in a clinical trial is the estimation of the difference between survival probabilities for the treatment groups at several points in time. As with the Wilcoxon and logrank tests, there is no well known nonparametric way to incorporate covariate adjustment into such estimation of treatment effects for survival rates. We propose a method that enables covariate adjustment for hypothesis testing with logrank or Wilcoxon scores. Related extensions for applying covariate adjustment to estimation of treatment effects are provided for differences in survival-rate counterparts to Kaplan Meier survival rates. The results represent differences in population average survival rates with adjustment for random imbalance of covariates between treatment groups. The methods are illustrated with a clinical trial example. PMID- 10379695 TI - On the utility of the Dirichlet distribution for meta-analysis of clinical studies. AB - Recently, there has been an increasing interest in combining efficacy (or safety) results from several clinical trials to draw an overall conclusion about the efficacy (or safety) of new investigational drugs. In a two-armed clinical trial, these efficacy outcomes are often expressed in terms of the two treatment effect means or the proportions of treatment success. For several clinical trials, efficacy assessment could be based on different types of clinical outcomes. These outcomes might be both categorical and noncategorical. Therefore, it is desirable to have a statistical model that is flexible enough for combining clinical trial data of both outcomes across several trials or studies. We discuss a simple statistical model with a potential for such flexibility. We illustrate the use of the model by providing a couple of numerical examples on the meta-analyses of efficacy data from prospective clinical trials/studies. PMID- 10379697 TI - Sample size for comparing linear growth curves. AB - Assuming a linear growth curve model under a suitable link function, we compute the sample size for comparing two treatment groups when the repeated measurements marginally follow exponential family distributions. From the treatment profiles of the chosen link function, we compute the common intercept beta0 and the regression slopes beta1 and beta2 to define delta = beta1 - beta2, the difference to be detected, under a specified alternative hypothesis. The dispersion matrices of the generalized estimating equations estimators are obtained under the null and alternative hypotheses using a suitable working correlation matrix. We compute the sample size assuming that delta is asymptotically normal. Details are worked out for repeated measures designs with binary and count data along with numerical examples. PMID- 10379698 TI - Multinomial phase II cancer trials incorporating response and early progression. AB - The objective of a phase II clinical trial in oncology is to assess the antitumor activity of a specific treatment regimen. A multiple-testing procedure is commonly used to decide whether the experimental treatment warrants further investigation based on patients' tumor response. There are ethical concerns about exposing patients to a new drug when the response rate is low and a relatively large number of patients have early progressive disease. Ensign et al. (1) proposed a stopping rule that rejects a drug early when there is a long run of early treatment failures. However, this approach may not be sensitive to pick up early progressors mixed with other nonresponders. In this paper, we present a multiple-stage stopping rule for a single-arm trial of an experimental treatment in which both tumor response and early progression are considered simultaneously. We use a multinomial model to accommodate an outcome of discrete multivariate responses in order to improve the efficiency of the stopping rule. The proposed multiple-testing procedure requires that both the numbers of responses and early progressions fall within the boundaries satisfying the stopping criteria in order to stop the study. Simulation is performed to validate these results and to compare them with other commonly used designs. Other applications of this method are also discussed. PMID- 10379699 TI - Noninferiority testing in crossover trials with correlated binary outcomes and small event proportions with applications to the analysis of condom failure data. AB - Regulatory agencies may require demonstrating a new therapy or device as noninferior to an existing method. For example, a new condom type must demonstrate noninferiority to existing latex condoms before consideration as an equivalent method of pregnancy prevention. Studies designed to assess condom effectiveness typically measure experimental and standard condom failure in a crossover trial, resulting in unbalanced correlated binary outcomes with low event proportions. We used simulations to evaluate the test size of a simple population average approach to noninferiority testing with the small event proportions, intracluster correlations, and sample sizes frequently found in condom studies. Results emphasize the importance of considering test accuracy when designing any study. PMID- 10379700 TI - Skin diseases related to abnormality in desmosomes and hemidesmosomes--editorial review. PMID- 10379701 TI - Hereditary diseases of desmosomes. AB - Desmosomes are key adhesion complexes in most epithelia, including epidermis. Although structural components of desmosomes have been identified as target antigens in several of the autoimmune blistering skin diseases, there are relatively few data on inherited disorders arising from mutations in genes encoding these proteins and glycoproteins. For example, an association between an inherited abnormality of desmosomes and Darier disease and Hailey Hailey disease has been proposed on histopathological grounds, but genetic linkage studies have not invoked known desmosomal gene loci. However, linkage analyses have implicated one or more of the desmosomal cadherins (desmogleins 1-3, desmocollins 1-3), the genes for which are tightly clustered within a 650-kb region on 18q12.1, in the pathogenesis of a different autosomal dominant genodermatosis, striate palmoplantar keratoderma. In addition, a rare autosomal recessive skin fragility ectodermal dysplasia syndrome has recently been recognised which results from total ablation of plakophilin 1, an intracellular desmosomal plaque protein that reinforces adhesion between the cytoskeleton and the cell membrane in terminally differentiating keratinocytes. In the future, it is likely that a number of other desmosome genodermatoses will be identified, each resulting from dominant or recessively inherited mutations in component structural proteins. PMID- 10379702 TI - Autoimmunity against desmosomal cadherins in pemphigus. AB - Pemphigus is a unique and interesting autoimmune disease, in which autoantibodies play a major pathogenic role and cause blister formation. Several questions raised from clinical observation in pemphigus have been answered with logic at the molecular level owing to recent remarkable progress in research in the field of pemphigus. The clinical phenotype of classic pemphigus, pemphigus vulgaris (PV) and pemphigus foliaceus (PF), is defined by anti-desmoglein autoantibody profile. Sera containing anti-Dsg3 IgG alone cause mucosal dominant PV with limited skin involvement. Sera containing both anti-Dsg3 and anti-Dsg1 cause mucocutaneous PV, which affects both the skin and mucous membrane. Sera containing only anti-Dsg1 cause PF, which shows cutaneous but no mucosal involvement. In herpetiform pemphigus (HP) most sera recognize Dsg1 and the rest of them recognize Dsg3, indicating that HP is a clinical variant of PF or PV. Patients with paraneoplastic pemphigus (PNP) have autoantibodies against multiple molecules. Now we know that they have autoantibodies against all members of the plakin family, which are cytoplasmic proteins and include desmoplakin, BPAG1, envoplakin, periplakin, and plectin. Cell surface target antigens of PNP, which blister-inducing pathogenic autoantibodies attack, were finally discovered to be Dsg3 and Dsg1. Therefore, PNP is characterized as an autoimmune disease against plakin molecules and desmogleins. Autoimmune targets of IgA pemphigus are likely more heterogeneous than originally thought. So far, desmocollin 1, Dsg3, and Dsg1 are known as their target antigens. Thus, pemphigus has become one of well characterized tissue-specific autoimmune diseases. Pemphigus will be a good model disease in the next century to address the central issue of autoimmune disease and basic immunology; why and how do patients with autoimmune diseases start to recognize self as non-self? PMID- 10379703 TI - Hereditary skin diseases of hemidesmosomes. AB - Studies of hereditary blistering skin diseases (epidermolysis bullosa) and targeted gene mutation experiments in knockout mice have greatly improved our understanding of hemidesmosomes and their associated structures in the cytoskeleton and basement membrane of the skin and mucous membranes. At least 10 molecules are recruited in hemidesmosome complexes, where they interact in a complex way. Hemidesmosomes are not simple adhesion devices, but also transduce signals for cell spreading, cell proliferation and basement membrane organisation. The dynamics of a hemidesmosome raises the metaphor of a self assembling suspension bridge which evokes activities on both sides of the river. This review summarises our current knowledge of the molecular pathology of hemidesmosomes caused by hereditary skin disease or gene targeting experiment. PMID- 10379704 TI - Hereditary skin diseases of anchoring fibrils. AB - Remarkable progress has been made in the last few years in understanding the functions of the anchoring fibrils, polymers of collagen VII, that connect the epidermal basement membrane with the dermal connective tissue. Novel insights into the biology of these fibrils have been gained from studies on dystrophic epidermolysis bullosa (DEB), a group of inherited blistering disorders caused by abnormalities of the anchoring fibrils. Mutations in the COL7A1 gene encoding collagen VII have been disclosed in a number of DEB families, and the mutation analyses and studies on genotype-phenotype correlations in DEB have revealed an unusual complexity of the gene defects and their biological consequences. In analogy to heritable disorders of other collagen genes, predictable phenotypes of COL7A1 mutations causing premature termination codons (PTC) or dominant negative interference have been observed. However, collagen VII seems to be unique among collagens in that many mutations lead to minimal phenotypes, or to no phenotype at all. Furthermore, the mild DEB phenotypes can be severely modulated by a second mutation in individuals compound heterozygous for two different COL7A1 defects. Therefore, not only definition of mutations with diagnostic analyses, but also cell biological, protein chemical and suprastructural studies of the mutated molecules are required for understanding the pathomechanisms underlying DEB. PMID- 10379705 TI - Acquired skin disease of hemidesmosomes. AB - The hemidesmosome is a membrane-associated supramolecular dermal epidermal complex linking the cytoskeleton of the basal keratinocyte to structures within the papillary dermis. Different components of this complex have been identified as autoantigens in autoimmune bullous skin diseases. Some of the autoantigens have been characterized at the molecular level. Little is known, however, about the factors that initiate the production of autoantibodies. By histopathology, acquired skin diseases of hemidesmosomes show subepidermal blisters and by direct immunofluorescence, linear deposits of IgG, C3 or IgA at the dermal epidermal junction. Bullous pemphigoid (BP) is the most common acquired disease of hemidesmosomes. Two proteins, BP180 and BP230, have been identified as primary targets of autoantibodies in BP. In addition, pemphigoid/herpes gestationis, lichen planus pemphigoides, cicatricial pemphigoid and linear IgA disease are characterized by an immune response to BP180. Laminin 5 is another well characterized anchoring filament-lamina densa component of hemidesmosomes. Patients with autoantibodies to laminin 5 show the clinical phenotype of cicatricial pemphigoid. Other acquired skin diseases of the hemidesmosomes reveal autoantibodies to a plectin-like protein, the beta4 subunit of alpha6beta4 integrin, uncein and a not yet characterized 168 kDa protein. Recently, diseases with autoantibodies to 105 and 200 kDa proteins of the lower lamina lucida have been reported. The association of these autoantigens with hemidesmosomes still needs to be demonstrated. Finally, anchoring fibrils associate with the dermal epidermal anchoring complex. The major structural component of anchoring fibrils is type VII collagen, the autoantigen of epidermolysis bullosa acquisita. PMID- 10379706 TI - Attacking cataract blindness. PMID- 10379707 TI - There is no use crying over spilt tears: the surgical management of primary acquired nasolacrimal duct obstruction. PMID- 10379708 TI - Ophthalmic interventions in the Developing World: insights for successful outcomes. PMID- 10379709 TI - Localized scotomata detected with temporal modulation perimetry in central serous chorioretinopathy. AB - AIM: Flicker deficits have been reported in various maculopathies, including age related macular degeneration. We test whether flicker losses exist in patients with central serous chorioretinopathy (CSC) and whether the size and flicker frequency of the target is important in detecting such losses. METHODS: We examined four CSC patients with temporal modulation (flicker perception) perimetry using the Medmont auto-flicker module (Medmont Pty Ltd, Melbourne, Vic. Australia), as well as static perimetry and colour vision. One case was examined using sophisticated laboratory equipment to precisely measure their temporal contrast sensitivity function (temporal CSF or de Lange curve) using larger targets to consider the effect of target frequency and size. Two patients were followed longitudinally and tested after resolution of the maculopathy. We compared our patients with an age-matched control group of 11 people. RESULTS: Temporal modulation perimetry detected larger and more localized defects in all cases of active CSC compared with static perimetry. There appeared to be size and frequency tuning to the deficit, with greatest loss being found at 16 Hz with small (0.5 degree) targets. The losses resolved in one case where the retina recovered in 4 weeks, but remained to a lesser degree in another case who suffered a 2 year long fluctuating course before the CSC subsided. CONCLUSIONS: Temporal modulation perimetry detects a loss of flicker sensitivity in patients with CSC. Deeper and more clearly defined scotomata are found with a flickering stimulus compared with a steady state one. The greatest losses of flicker sensitivity are found with 16 Hz modulation and with small targets located directly over the lesion. The duration of the disease may be important for recovery of flicker sensitivity. Temporal modulation perimetry appears to be a valuable tool for the confirmation of functional loss due to CSC. PMID- 10379710 TI - Localization of IgG in the normal and dystrophic rat retina after laser lesions. AB - PURPOSE: To test the hypothesis that access to extravasated plasma protein IgG may influence photoreceptor survival following laser photocoagulation and to determine whether this correlates with the retinal glial reaction. METHODS: A total of 45 rats (18 Royal College of Surgeons (RCS) dystrophic and 18 RCS-rdy+ congenic control) were used for this experiment. Nine non-lasered littermates of same age were used as controls. The superior retinas of postnatal day 23 rats were irradiated with a grid pattern of 40 argon green laser lesions of 50 microm in diameter and two powers (150 and 300 mW) for 0.2 s. At various times after laser lesions (up to 14 days), animals were perfused, the retinas snap frozen and sectioned on a cryostat. A one-step immunohistochemical technique was used by incubating with rabbit anti-rat IgG conjugated directly to horseradish peroxidase. Adjacent sections were processed using an antibody to glial fibrillary acidic protein (GFAP) by the standard avidin-biotin complex method. RESULTS: The labelling pattern for extravasated IgG after laser lesion was very similar in both RCS and RCS-rdy+ rat retinas. At 6, 12 and 24 h after lesions, IgG immunoreactivity (IR) was very intense in the lesion core and flanks. The outer plexiform layer (OPL) and photoreceptor inner segments provided a ready pathway for lateral spread of IgG. However, in the outer nuclear layer (ONL), IgG localization was much more restricted. Despite very intense IgG IR in the ONL of the coagulated lesion core, there was always a very sharply delineated boundary where the label abruptly halted. The GFAP labelling in both RCS dystrophic and RCS-rdy+ congenic control rat retinas showed that this boundary was between normal and necrotic cells because there was a core where GFAP was not produced by Muller cells. By 2 days after lesions, the coagulated cells in the lesion core were being removed by phagocytic cells that were IgG IR. Labelled phagocytic cells were also found among the inner and outer segment region on the lesion flanks. There was still IgG IR in the lesion, but the label was faint. No IgG IR was found in the retina at 3, 4, 7 and 14 days after lesions. Absorption control with pure rat IgG showed the label to be specific. CONCLUSIONS: The extravasated IgG was derived from the choroidal circulation because at no stage was IgG localized around the retinal vasculature. The IgG labelling was surprisingly widespread and, therefore, did not correlate with photoreceptor sparing, although it preceded the widespread Muller cell expression of GFAP and may, therefore, trigger glial reaction. PMID- 10379711 TI - A new model of orthotopic penetrating corneal transplantation in the sheep: graft survival, phenotypes of graft-infiltrating cells and local cytokine production. AB - BACKGROUND: Orthotopic penetrating keratoplasty in the sheep was developed as an outbred preclinical model to allow correlation of the cellular infiltrate during graft rejection with local production of cytokine mRNA. METHODS: Penetrating corneal autografts and allografts were performed in Merino sheep. Graft outcome was followed at the slit-lamp. Corneal infiltrates were examined by immunoperoxidase staining on postmortem specimens. Cytokine mRNA was detected by polymerase chain reaction. RESULTS: Corneal autografts survived indefinitely. Allografts became vascularized and underwent rejection at a median of 20 days postgraft. Both endothelial and epithelial rejection lines were observed. Immunohistochemical staining of rejecting grafts showed up-regulation of major histocompatibility complex class I molecules on corneal graft epithelium, damaged or absent graft endothelium and a marked, predominantly mononuclear cell infiltrate. CD4-positive T cells were observed in the graft within 2 days of the onset of rejection, followed several days later by CD8-positive T cells. Messenger RNA transcripts for interleukin (IL)-2, tumour necrosis factor (TNF) alpha and IL-10 (but not for interferon (IFN)-gamma or IL-4) were found in autografted corneas. Proportionately, more allografts than autografts contained transcripts for IL-2 and TNF-alpha, and IFN-gamma was detected in three of four allografts. CONCLUSIONS: Corneal graft rejection in the sheep is macroscopically and histologically similar to human corneal graft rejection. Allografts become infiltrated by both CD4- and CD8-positive T cells and local production of pro inflammatory cytokines occurs during graft rejection. PMID- 10379712 TI - Number needed to treat: a useful new method of assessing the magnitude of treatment effect and its application to the management of diabetic retinopathy. AB - The magnitude of treatment effect from randomized controlled trials can be measured in various ways. The number needed to treat is a recently described measure that has been shown to offer several advantages in the clinical interpretation and application of reported treatment effects. It quantifies the number of patents that must be treated in order to prevent one patient from developing the specified outcome. The methods necessary to calculate traditional measures of treatment effect, as well as the number needed to treat, are outlined using the management of diabetic retinopathy as an example. The uses and limitations of the number needed to treat are also discussed. PMID- 10379713 TI - Corneal laceration and forehead erythema: a syndrome in infants. AB - PURPOSE: To illustrate a syndrome of transient erythema in the first division of the fifth cranial nerve caused by minor corneal laceration, occurring in infants. METHODS: Case reports are presented, together with photographs. These must, of necessity, usually be obtained by the general practitioner because of the short duration of the signs. RESULTS/CONCLUSIONS: Opinion is sought as to whether other practitioners have recognized this association and whether it has been described elsewhere. PMID- 10379714 TI - Unusual presentations of pleomorphic adenoma and adenoid cystic carcinoma of the lacrimal gland. AB - PURPOSE: To report two atypical cases of pleomorphic adenoma and adenoid cystic tumours of the lacrimal gland. METHODS: Two case reports are presented. The first is of a 65-year-old female with a long history of right hypoglobus with sudden recent worsening. Computed tomography (CT) showed a round, well-defined lesion in the fossa for the lacrimal gland with an anterior hypodense extension suggestive of possible malignancy in a pleomorphic adenoma. The tumour in the second case, a 35-year-old male, was diagnosed after presentation following a relatively minor periorbital injury. The smooth rounded mass on CT scan was suggestive of a benign lacrimal gland tumour. RESULTS: The lesion in case 1 was excised with a diagnosis of haemorrhage within a pleomorphic adenoma. The lesion in case 2 was excised with a diagnosis of adenoid cystic carcinoma of the lacrimal gland with pseudocapsule. CONCLUSIONS: Haemorrhagic cyst developing beneath the pseudocapsule of a pleomorphic adenoma should be considered in the differential diagnosis of secondary development of malignancy in a pleomorphic adenoma. Adenoid cystic tumours of the lacrimal gland can present with a pseudocapsule. PMID- 10379715 TI - Squamous cell carcinoma with necrotizing scleritis. AB - PURPOSE: To report on a case of limbal squamous cell carcinoma (SCC) with necrotizing scleritis in a young, previously healthy, white Australian male. METHODS: A 31-year-old man presented with a left limbal lesion intermittently causing a red eye and foreign body sensation. He had enjoyed surfing for many years. Repeat HIV tests were negative and the lesion was biopsied. RESULTS: Biopsy showed a well-differentiated SCC apparently arising in an intra-epithelial (in situ) carcinoma of the conjunctiva. The lesion was excised and a corneoscleral graft repair was performed. CONCLUSION: The present case highlights the potential for a significant increase in the prevalence of ocular surface neoplasia in healthy young people who have had excessive UV-B exposure. PMID- 10379716 TI - Conservative management of documented neuroretinitis in cat scratch disease associated with Bartonella henselae infection. AB - BACKGROUND: Bartonella henselae has been identified as the causative agent of the neuroretinitis associated with cat scratch disease (CSD). Immunofluorescent antibody tests with good sensitivity and specificity are available to aid in diagnosis. Despite diagnostic advances, optimal management remains controversial. We present a case of documented B. henselae macular neuroretinitis managed without antibiotics and discuss antibiotic use in this condition. METHODS: We examined a young woman with macular neuroretinitis and established a diagnosis of CSD. Management consisted of a review of the literature, followed by educating her about the condition and close observation. We documented the course of her disease. RESULTS: We diagnosed neuroretinitis associated with B. henselae infection based on immunofluorescent antibody titres and clinical presentation. Our patient's neuroretinitis resolved promptly without antibiotic therapy. CONCLUSIONS: Macular neuroretinitis in CSD can be satisfactorily diagnosed with the use of fluorescent antibodies in the appropriate clinical setting. Optimal treatment for the disease has not been established and observation combined with patient education remains an appropriate option. The self-limited nature of the disease implies that treatment studies not using controls must be interpreted with great caution. Adverse drug reactions and other iatrogenic complications can be reduced by limiting antibiotic use in settings where a meaningful treatment benefit has not been established. PMID- 10379717 TI - Anterior ischaemic optic neuropathy in a patient with optic disc drusen. AB - BACKGROUND: Although visual field defects are well-known complications of optic disc drusen, reduction in visual acuity with this condition is rare. METHOD/RESULTS: We report on a 68-year-old male with bilateral optic disc drusen who presented with monocular loss of vision in the right eye associated with an inferior altitudinal visual field defect and signs consistent with acute anterior ischaemic optic neuropathy, confirmed on fluorescein angiography. He also had a left inferior nasal step, but no evidence of glaucomatous cupping. The disc drusen were documented clinically and on B scan ultrasound and computed tomography. CONCLUSIONS: The diagnosis of acute anterior ischaemic optic neuropathy should be considered in patients with optic disc drusen who present with reduced visual acuity, particularly when the visual loss has been acute and non-progressive and is associated with altitudinal field loss and characteristic fluorescein angiography signs. PMID- 10379718 TI - Cataract, cost: curious questions. PMID- 10379719 TI - Irritability following traumatic brain injury. AB - This study was undertaken to identify the clinical and pathoanatomical correlates of irritability in patients with closed head injuries. A consecutive series of 66 patients was assessed in hospital and at 3, 6, 9, and 12-month follow-ups. Patients fulfilling criteria for irritability were divided into 2 groups based on the immediate or delayed onset of their irritability and compared with patients without irritability for background characteristics, impairment variables, and lesion characteristics. There were 12 patients (18.2%) with acute onset irritability and 10 (15.1%) with delayed onset irritability. Acute onset irritability patients had a higher frequency of left cortical lesions. Delayed onset irritability patients showed a strong association with poor social functioning and greater impairment in activities of daily living. The findings suggest that post-brain injury irritability may have different causes and treatment in the acute and chronic stages. PMID- 10379720 TI - Compulsive physical activity in adolescents with anorexia nervosa: a psychobehavioral spiral of pathology. AB - The excessive exercising that is frequently observed in anorexia nervosa (AN) has been viewed both as an addictive behavior and as a type of obsessive compulsive disorder. The present study tested a nonrecursive structural equation model that specified associations among personality factors, cognitions, and behavior in the development and progression of excessive exercise in adolescent patients with AN. As proposed, findings indicated that both addictive personality and obsessive compulsive personality contributed to excessive exercising by means of their influence on obligatory/pathological cognitions about exercising. Childhood physical activity also predicted excessive exercising. The implications of these results are discussed from a psychobiological perspective. PMID- 10379721 TI - Posttraumatic stress disorder symptoms related to psychosis and acute involuntary hospitalization in schizophrenic and delusional patients. AB - The aims of this study were: a) to assess the prevalence of posttraumatic stress disorder (PTSD) after an acute psychotic episode in schizophrenic and delusional patients, b) to explore which psychotic symptoms and aspects of treatment were associated with traumatization, and c) to compare the extent of the traumatic impact of psychosis and involuntary hospitalization. Forty-six schizophrenic and delusional patients were assessed with the Positive and Negative Syndrome Scale (PANSS), the Impact of Event Scale-Revised (IES-R), and the Clinician Administered PTSD Scale (CAPS) at weeks 1 and 8 after acute psychiatric admission. Traumatic symptoms related to psychosis and coercive measures were scored separately. The prevalence of PTSD was found to be 11%. Sixty-nine percent of traumatic symptoms were related to psychosis and 24% to hospitalization. High PANSS score at week 8 was the strongest risk factor for the development of PTSD. Particularly positive and depressive/anxious symptomatology were associated with psychosis-related traumatic symptoms at both weeks 1 and 8. These data suggest that, in general, schizophrenic and delusional symptoms are more traumatic than the coercive measures used to control them. PMID- 10379722 TI - Combat exposure, posttraumatic stress disorder symptoms, and health behaviors as predictors of self-reported physical health in older veterans. AB - We used path analysis to model the effects of combat exposure, posttraumatic stress disorder (PTSD) symptoms, and health behaviors on physical health. Participants were 921 male military veterans from the Normative Aging Study. Their mean age at time of study was 65. Measures of combat exposure, PTSD symptoms, smoking, and alcohol problems were used to predict subsequent self reported physical health status. Both combat exposure and PTSD were correlated with poorer health. In path analysis, combat exposure had only an indirect effect on health status, through PTSD, whereas PTSD had a direct effect. Smoking had a small effect on health status but did not mediate the effects of PTSD, and alcohol was unrelated to health status. We conclude that PTSD is an important predictor of physical health and encourage further investigation of health behaviors and other possible mediators of this relationship. PMID- 10379723 TI - Symptom-based predictors of a 10-year chronic course of treated depression. AB - The high likelihood of a chronic course of depression underscores the need to identify at intake patients most at risk for long-term nonremission. In a naturalistic study of 313 unipolar depressed patients, potential symptom-based risk factors were assessed at treatment intake and were used to predict a chronic course of treated depression over a 10-year interval. The prototypic chronically depressed patient was an individual who at baseline experienced more severe symptoms of fatigue, loss of interest in usual activities, trouble sleeping, and thoughts about death or suicide; was not calm, successful, or self-confident; and did not socialize with friends outside the home, and frequently coped with stressors by avoiding other people. A larger number of risk factors was associated with a higher likelihood of experiencing a chronic course. High-risk patients who received more psychological treatment during the index episode were more likely to experience a long-term course of remission or partial remission. PMID- 10379725 TI - The relationship between insight and suicidality among patients with schizophrenia. PMID- 10379724 TI - Application of the high risk model of threat perception to a primary care patient population. AB - This study was designed to test the hypothesis that patients with a tendency to somatize psychological distress into physical symptoms could be differentiated from patients who do not somatize on the basis of specific predisposing factors defined by the High Risk Model of Threat Perception. Patients in a family practice were assessed for the tendency to somatize by the Diagnostic Interview Schedule (DIS) and by physician rating. Twenty-seven percent of the patients were positive for tendency to somatize by physician rating. These patients had relatively high negative affect, absorption, catastrophizing, self-reported pain and stress, and greater utilization of services. None of the patients assessed by the DIS met criteria for somatization disorder, but 28% were positive for somatoform pain disorder. These patients also scored higher on the negative affect questionnaire, tended to have higher absorption scores, reported greater pain and stress, and utilized more services. Results of this study are partially supportive of the High Risk Model of Threat Perception, because two of the predisposer factors were associated both with tendency to somatize by physician rating and with somatoform pain disorder by interview. The higher utilization of services in the somatizing patients has cost and service ramifications. Treatment of patients with tendencies to somatize within a family practice setting are discussed. PMID- 10379726 TI - Predictors of participation in campaigns against mental illness stigma. PMID- 10379727 TI - Theory of mind and the delusional disorders. PMID- 10379728 TI - Intravesical capsaicin and resiniferatoxin therapy: spicing up the ways to treat the overactive bladder. AB - PURPOSE: Pharmacological treatment of the overactive bladder relies on partially blocking the efferent parasympathetic innervation to the detrusor with anticholinergic drugs. However, often these drugs have troublesome side effects and doses are insufficient to restore continence in patients with detrusor instability. We present the background, basic and clinical research with intravesical instillation of capsaicin and resiniferatoxin as treatments for the overactive bladder. MATERIALS AND METHODS: Capsaicin, the main pungent ingredient in hot peppers of the genus Capsicum, is a specific neurotoxin that desensitizes C fiber afferent neurons which may be responsible for signals that trigger detrusor overactivity. RESULTS: In the last 6 years studies have demonstrated encouraging improvement in lower urinary tract symptoms with minimal long-term complications. Most of these studies have also demonstrated that the acute pain and irritation associated with capsaicin are major deterrents to widespread use. Therefore, resiniferatoxin, an ultra-potent analogue of capsaicin which appears to have similar efficacy but less acute side effects, may be more useful. CONCLUSIONS: Intravesical capsaicin and resiniferatoxin are novel and promising treatments for the overactive bladder, with profound basic and clinical implications. PMID- 10379729 TI - BCL-2 and p53 expression in clinically localized prostate cancer predicts response to external beam radiotherapy. AB - PURPOSE: Clinicians have long been hampered by the inability to distinguish patients with localized prostate cancer who will and will not respond to radiotherapy. In a significant proportion of patients therapy fails as determined by increasing posttreatment serum prostate specific antigen (PSA). We evaluated the expression of 2 key regulators of apoptosis, bcl-2 and p53, relative to treatment outcomes in patients who received external beam radiotherapy for clinically organ confined carcinoma of the prostate. MATERIALS AND METHODS: Immunohistochemical staining for bcl-2 and p53 on pretreatment needle biopsies was performed in 54 patients who were treated with radiotherapy for localized prostate cancer. Expression was scored using strict criteria. Nadir PSA less than 1 ng./ml. after therapy was considered a successful treatment response. RESULTS: There was a predominance of stage T1c cancer (74%) with a mean Gleason score of 6.9 and an average pretreatment PSA of 25.3 ng./ml. Overall 54% of the patients did not have a nadir PSA of less than 1 ng./ml. Of the bcl-2 positive cases therapy ultimately failed in 85%. Similarly 88% of the patients with p53 positive biopsies had treatment failure and in all with bcl-2 as well as p53 expression radiotherapy failed. Expression of bcl-2 and p53 was an independent prognostic variable for treatment failure with odds ratios (95% confidence interval) of 7.3 and 10.8, respectively. CONCLUSIONS: Expression of bcl-2 and p53 was associated with treatment failure after external beam radiation therapy. These findings suggest that bcl-2 and p53 expression in pretreatment biopsies may be helpful for predicting response to definitive radiotherapy. PMID- 10379730 TI - Petroleum jelly is an ideal contact medium for pain reduction and successful treatment with extracorporeal shock wave lithotripsy. AB - PURPOSE: Various minimally invasive approaches to reduce pain during extracorporeal shock wave lithotripsy (ESWL) have been described. We compared petroleum jelly (Vaselinet) and ultrasound gel in vitro as a contact medium based on the stone fragmentation rate. The analgesic effect of cutaneous petroleum jelly was tested against eutectic mixture of local anesthesia. We also evaluated the outcome of ESWL in a large group of patients treated with petroleum jelly. MATERIALS AND METHODS: In vitro 3 artificial stones were completely fragmented with a MFL 5000* lithotriptor using petroleum jelly or ultrasound gel as a contact medium. A total of 110 patients (group 1) received petroleum jelly before treatment with the same lithotriptor. After retrospective analysis of group 1 we matched 32 patients (group 2) receiving cutaneous eutectic mixture of local anesthesia. Because of the favorable results with petroleum jelly, we used it in another 148 patients, for a total of 258 patients (group V). Treatment dependent pain was scored using a questionnaire as 1--no, 2--minor, 3--tolerable and 4- intolerable. ESWL without additional analgesics had a pain score of 1 to 3. RESULTS: In vitro petroleum jelly had a superior fragmentation rate compared to ultrasound gel. Our long-term experience with the lithotriptor indicated that only 30% of patients required no additional analgesics with cutaneous ultrasound gel. In contrast, no additional analgesics were needed in only 38% of group 2 compared to 81.8% of group V. The stone fragmentation rate did not differ statistically between groups. CONCLUSIONS: Cutaneous petroleum jelly offers a noninvasive, highly effective, inexpensive treatment modality with no side effects and significant reduction in pain. This ointment is our contact medium of choice. PMID- 10379731 TI - Sensitivity of noncontrast helical computerized tomography and plain film radiography compared to flexible nephroscopy for detecting residual fragments after percutaneous nephrostolithotomy. AB - PURPOSE: We prospectively compared plain film radiography and noncontrast, thin cut helical computerized tomography (CT) to flexible nephroscopy for detecting residual stones after percutaneous nephrostolithotomy. MATERIALS AND METHODS: We prospectively evaluated 36 patients (41 renal units) undergoing percutaneous nephrostolithotomy for large (greater than 3 cm., 23 renal units) or staghorn (18 renal units) calculi. All patients underwent postoperative imaging with plain film of the kidneys, ureters and bladder and noncontrast helical CT, and flexible nephroscopy on postoperative day 2 or 3. The size and location of residual fragments determined radiographically and identified by flexible nephroscopy were compared. RESULTS: Plain film radiography and CT detected an average of 0.7 and 3.4 stones per renal unit, respectively. With a mean operating time plus or minus standard deviation of 77.3+/-35 minutes and a mean fluoroscopy time of 7.6+/-6.7 minutes, an average of 2.3 stones per renal unit were retrieved at flexible nephroscopy. In 90.2% of renal units all calices could be directly inspected. The sensitivity and specificity were 46% and 82% for plain film radiography, and 100% and 62% for CT, respectively, using flexible nephroscopy as the gold standard for detecting residual stones. The overall stone-free rate after flexible nephroscopy was 92.6%. The cost of this procedure is $5,625.13 compared to $220 for CT, including the interpretation fee, at our institution. CONCLUSIONS: Selective use of flexible nephroscopy after percutaneous nephrostolithotomy based on positive CT findings will avoid an unnecessary operation in 20% of patients. The rate of unnecessary procedures is 32% if all patients undergo flexible nephroscopy, regardless of radiographic findings. At our institution this strategy will result in a cost savings of $109,687 per 100 patients. PMID- 10379732 TI - A prospective study of recurrence rate and risk factors for recurrence after a first renal stone. AB - PURPOSE: We investigate further the recurrence rate and risk factors for recurrence in 300 consecutive patients who presented to our stone clinic after a first stone episode 7 to 17 years ago. MATERIALS AND METHODS: The medical records of the patients who presented consecutively with a first stone episode from 1980 to 1990 were studied and supplemented by a followup mail questionnaire and telephone interviews. At first visit serum samples were taken from all patients and 24-hour urine samples were collected for metabolic testing. RESULTS: A total of 195 patients were followed successfully, of whom 52 (27%) experienced symptomatic stone recurrence after a mean plus or minus standard deviation of 7.5+/-5.9 years. However, ultrasound examination of 36 symptom-free patients showed recurrent stones in 28%. Comparison of patients with or without recurrence confirmed that recurrence was not influenced by sex, family history of stones and urinary risk factors. However, age at onset of the disease was lower for patients who had 2 or more stones during followup than those who had only 1 stone or no recurrence. CONCLUSIONS: Stones can recur as long as 10 years after the first episode, although the rate is lower than previously reported. The metabolic evaluation after a first stone episode needs to be reappraised in terms of its cost-effectiveness, since recurrences do not seem to be predictable from standard laboratory tests. PMID- 10379733 TI - Helical computerized tomography arteriography for evaluation of live renal donors undergoing laparoscopic nephrectomy. AB - PURPOSE: Traditionally, live renal donors are evaluated with excretory urography and renal arteriography. Helical computerized tomography (CT) arteriography offers a less invasive alternative for demonstrating necessary anatomical information before laparoscopic allograft harvest. We evaluate the accuracy of helical CT arteriography in depicting renal vascular anatomy with an emphasis on the detection of arterial and venous anomalies. MATERIALS AND METHODS: Imaging studies were done on 175 patients according to a standard CT arteriography protocol with early arterial phase scanning (14 to 20-second delay), and 1 mm. axial and 3-dimensional maximum intensity projection reconstructions. Renal vascular anatomy was mapped with attention to aberrant arterial and venous anatomy. Intraoperative findings were correlated at laparoscopic donor nephrectomy. RESULTS: There was overall agreement between CT arteriography and laparoscopic findings in 163 cases (93%). Supernumerary renal arteries were identified in 40 cases (23%). Sensitivity, specificity and accuracy of CT arteriography for arterial anatomy were 91, 98 and 96%, respectively. Cases with less than 2 mm. accessory arteries or early branching single vessels simulating dual arteries were misdiagnosed. Venous anomalies occurred in 11 patients (6.3%). Sensitivity, specificity and accuracy of CT arteriography for venous anatomy were 65, 100, and 97%, respectively. Misdiagnoses included early venous bifurcations and supernumerary tributary veins, which were poorly opacified. CONCLUSIONS: Helical CT is highly accurate and specific for the demonstration of renal arterial anatomy. Poor opacification resulted in a lower sensitivity for venous anatomy. Overall, helical CT provides essential anatomical information, and is an alternative to standard urography and arteriography. PMID- 10379734 TI - Transperitoneal and retroperitoneal laparoscopic nephrectomy for giant hydronephrosis. AB - PURPOSE: We evaluate laparoscopic nephrectomy for giant hydronephrosis with an emphasis on the operative technique of retroperitoneoscopic surgery. MATERIALS AND METHODS: During the last 2 years 13 men and 5 women underwent laparoscopic nephrectomy for giant hydronephrosis via a transperitoneal (6) or retroperitoneal (12) approach. The etiology was congenital ureteropelvic junction obstruction in 17 patients and hydronephrosis caused by stone disease in 1. Three patients had a contralateral obstructed kidney. Renal parameters were normal in all patients. RESULTS: All procedures were successfully completed without the need for conversion to open surgery. Mean operating time was 113.8 minutes (range 70 to 165) and average blood loss was 260 ml. (range 40 to 600). No patient required a blood transfusion. Postoperative recovery was uneventful with an average postoperative hospital stay of only 3.2 days (range 2 to 5). CONCLUSIONS: Laparoscopic nephrectomy is a good alternative to open surgery for giant hydronephrosis and significantly reduced the morbidity of surgery. A retroperitoneal approach is feasible, despite the large amount of retroperitoneal space occupied by these hugely dilated kidneys. Modifications of our technique have been invaluable to the successful outcome in this series. PMID- 10379735 TI - Renal oncocytoma: multifocality, bilateralism, metachronous tumor development and coexistent renal cell carcinoma. AB - PURPOSE: We analyzed a large series of cases of renal oncocytoma to define the incidence of coexistent renal cell carcinoma, multifocality, bilateralism and metachronous tumor development. MATERIALS AND METHODS: Between 1980 and 1997, 100 men and 38 women with a mean age of 68 years with oncocytoma, were treated surgically at our institution. We analyzed tumor characteristics and reviewed specimens for coexistent renal cell carcinoma. RESULTS: Tumors were discovered incidentally in 58% of the cases. Specimens were obtained from 84 radical and 70 partial nephrectomies. Tumor size ranged from 0.3 to 14.5 cm. (median 3.2). Oncocytoma was unilateral in 131 cases (95%) and bilateral in 7 (5%), while there were multiple oncocytomas in 8 (6%). Mean followup was 41 months (range 0 to 200). The disease specific survival rate was 100% and no patient had metastasis. In 6 patients (4%) metachronous oncocytoma developed during followup. No patient had locally recurrent oncocytoma after partial nephrectomy for a solitary renal oncocytoma. Renal cell carcinoma and oncocytoma were found in 14 patients (10%), including unilateral synchronous disease in 9 and bilateral synchronous disease in 5. CONCLUSIONS: Our data support the benign nature of renal oncocytoma. Multifocality, bilateralism and metachronous tumor develop in approximately 4 to 6% of all cases. Renal cell carcinoma coexisted in 10% of oncocytoma cases. PMID- 10379736 TI - Interleukin-2 based immunotherapy for metastatic renal cell carcinoma with the kidney in place. AB - PURPOSE: We assessed morbidity, response and survival in patients with metastatic renal carcinoma treated with high dose intravenous interleukin-2 (IL-2) based immunotherapy with the primary renal tumor in place. MATERIALS AND METHODS: We retrospectively analyzed the records of patients with metastatic renal carcinoma and the primary kidney tumor in situ who were treated at the surgery branch of the National Cancer Institute. Of the patients 607 were treated with IL-2 based therapy. Patient age, sex, sites of extrarenal disease, morbidity, and response and survival rates were examined. RESULTS: From 1986 to 1996, 51 patients with the majority of disease at extrarenal sites were treated with the primary tumor in place. Treatment involved IL-2 based regimens, reflecting the evolution of immunotherapy at the National Institutes of Health. When evaluating only extrarenal sites, response was complete in 1 and partial in 2 of the 51 cases (6%). No responses were noted in the primary renal tumor. Three patients with responses at extrarenal sites underwent nephrectomy. The duration of response in these 3 cases was greater than 88, 11 and 4 months, respectively. Median survival in all 51 patients was 13 months (range 1 to 86). CONCLUSIONS: Select patients may be treated with IL-2 based immunotherapy with the primary renal tumors in place with morbidity. A randomized study is needed to assess the role of cytoreductive nephrectomy for treating metastatic renal cell carcinoma. PMID- 10379737 TI - Metastatic renal cell carcinoma with concurrent inferior vena caval invasion: long-term survival after combination therapy with radical nephrectomy, vena caval thrombectomy and postoperative immunotherapy. AB - PURPOSE: We report our experience using aggressive multimodal therapy in a high risk group of patients with metastatic renal cell carcinoma and concurrent inferior vena caval extension. MATERIALS AND METHODS: We retrospectively reviewed the records of all patients in our kidney cancer database who had metastatic renal cell carcinoma and tumor thrombus extension into the inferior vena cava at the initial diagnosis. Patients were included in the study if they underwent radical nephrectomy and inferior venal caval thombectomy, and immunotherapy was planned for the postoperative period. Tumor size and grade, metastatic sites, level of vena caval extension, surgical complications and overall survival were obtained from the medical records. The primary end point analyzed was overall survival. RESULTS: We identified 31 cases of metastatic renal cell cancer with extensive disease and vena caval extension. Of the patients 23% had an isolated lung metastasis, and 53% had metastasis in the lung and at other sites. The remaining patients had involvement primarily at nonpulmonary metastatic sites, including lymph node in 38%, soft tissue in 13%, liver in 29% and bone in 10%. Average blood loss during nephrectomy was 3,200 cc (median 2,100) and the rate of major complications was 12%. Of the patients 80% underwent the full course of surgery and postoperative immunotherapy. At a mean followup of 18 months (34 for survivors) 26% of the patients are alive. Actuarial overall 5-year survival of the group was 17%. Tumor thrombus level did not correlate with overall survival, while immunotherapy, tumor grade and metastatic site provided significant prognostic information. In patients with an isolated pulmonary metastasis the 5 year survival rate was 43%, while in those with low grade tumors it was 52%. CONCLUSIONS: In contrast to the poor results of surgery only in patients with renal cell carcinoma and concurrent inferior venal caval invasion, reasonable 5 year survival may be achieved after combined aggressive surgery and immunotherapy. Patients in whom metastasis was limited to the lungs and those with grade 1 to 2 tumors had a better prognosis. With careful planning and experienced immunotherapists therapy may be completed in the majority of this high risk group of patients. PMID- 10379738 TI - Limitations of routine spiral computerized tomography in the evaluation of bladder trauma. AB - PURPOSE: We evaluate the accuracy of spiral computerized tomography (CT) in diagnosing traumatic bladder rupture. MATERIALS AND METHODS: Medical records of 24 consecutive patients diagnosed with traumatic bladder rupture at our level 1 trauma center from 1993 to 1998 were retrospectively reviewed. Of the patients 15 underwent retrograde cystography and spiral CT of the abdomen and pelvis. The results of these imaging studies were compared. RESULTS: Retrograde cystography successfully diagnosed all cases of bladder rupture and correctly classified injuries confirmed surgically. Spiral CT successfully diagnosed 9 of 15 bladder ruptures (60%), and correctly classified 4 of 5 intraperitoneal (80%) and 6 of 11 extraperitoneal (55%) ruptures. CONCLUSIONS: Spiral CT is less accurate than retrograde cystography in diagnosing traumatic bladder rupture. PMID- 10379739 TI - Exclusion criteria enhance the specificity and positive predictive value of NMP22 and BTA stat. AB - PURPOSE: The limitation of current urinary tumor markers is the low specificity and positive predictive value, which clinically manifests as a high false positive rate. We analyzed the false-positive data of 2 urinary tumor markers, NMP22 and the BTA stat tests. We examined the clinical categories of the false positive results, established relative exclusion criteria, and recalculated the specificity and positive predictive value after using the exclusion criteria. MATERIALS AND METHODS: A total of 278 symptomatic patients who presented to a urology clinic were asked to submit a single voided urine sample. Each sample was divided into 3 aliquots of which 1 was stabilized with the NMP22 test kit stabilizer and assayed for NMP22, 1 was tested for BTA stat and 1 was sent for cytological examination. All patients subsequently underwent office cystoscopy and bladder biopsy if indicated. RESULTS: Of the 278 symptomatic patients 112 presented with microscopic hematuria, 77 gross hematuria and 89 chronic symptoms of urinary frequency or dysuria. Of 34 cases (12%) of histologically confirmed bladder cancer NMP22 detected 28 (82.4%), BTA stat 23 (67.7%) and cytology only 10 (29.4%). When atypical cytologies were considered positive, cytology then detected 19 cases (55.9%). Elevated NMP22 values were positive in 28 cases and false-positive in 44 for a specificity of 82% and a positive predictive value of 38.9%. Similarly, BTA stat test was positive in 23 cases and false-positive in 43 for a specificity of 82.4% and a positive predictive value of 34.9%. When atypical cytologies were considered positive, the specificity and positive predictive value were 93% and 55.9%. Greater than 80% of the false-positive results were clinically categorized as benign inflammatory or infectious conditions, renal or bladder calculi, recent history of a foreign body in the urinary tract, bowel interposition segment, another genitourinary cancer or an instrumented urinary sample. A category of "no known pathology" was included in analysis as a control. History of ureteral stents or any bowel interposition segment had a 100% false-positive rate. Exclusion of all 6 clinical categories improved the specificity and positive predictive value of NMP22 (95.6%, 87.5%) and BTA stat (91.5%, 69.7%), and was similar to urinary cytology. CONCLUSIONS: Awareness and exclusion of the categories of false-positive results can increase the specificity and enhance the clinical usefulness of NMP22 and BTA stat tests. Similarly, treating an atypical cytology as positive can enhance the sensitivity and usefulness of urinary cytology. PMID- 10379740 TI - Tumor localization and systemic absorption of intravesical instillation of radio iodinated iododeoxyuridine in patients with bladder cancer. AB - PURPOSE: We evaluated tumor uptake and systemic distribution of intravesically instilled iododeoxyuridine (IUdR) in patients with superficial bladder cancer. MATERIALS AND METHODS: We performed 24 intravesical instillation studies in 11 patients with a mean age of 71 years. Radio-iodinated IUdR was administered through a Foley catheter. Gamma camera imaging was done after instillation and after 5 to 7 bladder irrigations. Tumor uptake was estimated by region of interest analysis. Bladder biopsy samples and surgical tumor specimens were tested for acid insoluble (deoxyribonucleic acid incorporated) radioactivity. Blood samples were obtained and analyzed for systemic absorption. RESULTS: Imaging was positive in all patients with bladder cancer. Average tumor uptake plus or minus standard deviation was 0.185+/-0.120% of the instilled dose. Preferential uptake of IUdR in the tumor was observed in all 6 patients undergoing tissue analysis. The tumor-to-normal bladder ratio ranged from 3.2 to 74,000 (median 202). Systemic absorption of IUdR was minimal. Blood sample analysis performed after intravesical instillation in all 11 cases revealed an average uptake of 3.2x10(-5)% instilled dose per ml. (range 0.69x10(-5) to 6.7x10(-5)) in the systemic circulation. Instillation within 24 hours after transurethral bladder tumor resection in 5 cases resulted in a higher but not dangerous average systemic uptake of 7.3x10(-4)% instilled dose per ml. (range 1.3x10(-5) to 2.6x10(-3)). Instillation 1 to 4 weeks after transurethral surgery in 8 cases resulted in no increased systemic absorption with an average blood level of 3.4+/-1.8x10(-5)% instilled dose per ml. There was no detectable distribution of radioactivity into other organs, including the thyroid. We noted no evidence of systemic toxicity in the study. CONCLUSIONS: Intravesical instillation of radio-iodinated IUdR achieves selective localization in the bladder tumor with minimal uptake by the normal bladder and minimal systemic absorption. The use of intravesical IUdR therapy for bladder cancer appears to be promising and requires further study. PMID- 10379741 TI - Argyrophilic nucleolar organizer region in proliferating cell has a predictive value for local recurrence in superficial bladder tumor. AB - PURPOSE: It has been shown in many carcinomas that the proliferation rate and number of argyrophilic nucleolar organizer regions (AgNOR) are associated with tumor aggressiveness. However, in bladder tumor the significance of the correlation between the number of AgNOR and tumor behavior remains controversial. Therefore, it would be helpful if a new technique could be developed that would allow for more accurate AgNOR counting in association with tumor behavior. We established the simultaneous staining technique of AgNOR with Ki-67 labeling to reveal the significance of AgNOR count in superficial bladder tumor. MATERIALS AND METHODS: A total of 50 paraffin sections of superficial bladder tumor were stained with AgNOR and Ki-67 (MIB-1). The numbers of AgNORs in proliferating (MIB 1 positive) or resting (MIB-1 negative) cells were counted from a total of 100 nuclei. Correlations between MIB-1 associated AgNOR count and clinicopathological parameters were statistically analyzed. RESULTS: The AgNOR count in proliferating cells was significantly higher than that in resting cells (p<0.01), and the count significantly increased with tumor grade (p<0.01). Based on recurrence-free survival analyses the local recurrence rate was significantly higher in patients with high proliferating cell NOR but not for those with resting or whole cells. However, no AgNOR score helped to select patients at high risk for disease progression. CONCLUSIONS: Proliferating cell NOR had a predictive value for local recurrence in patients with superficial bladder tumor. PMID- 10379742 TI - p53 mutations in bladder tumors inactivate the transactivation of the p21 and Bax genes, and have a predictive value for the clinical outcome after bacillus Calmette-Guerin therapy. AB - PURPOSE: We analyze the relationship among p53 mutations, p21 and Bax activation as well as their clinical implication in clinical response to intravesical bacillus Calmette-Guerin (BCG) therapy in high grade bladder tumors. MATERIALS AND METHODS: We analyzed a prospective series of 60 superficial bladder tumors using functional assays in yeast which test the transcriptional competence of p53 and can be used to identify p21 and Bax status. BCG instillations were given after initial tumor resection to 26 patients with a high risk of bladder invasive disease (pT1G3 tumors in 24 and carcinoma in situ in 2). RESULTS: No p53 alteration was detected in cases of pTa tumors. In contrast, p53 mutations were detected in 16 of 24 patients (66%) with pT1 G3 tumors and in 2 with primary carcinoma in situ. These 18 mutant samples scored also mutant for transactivation of p21 and Bax reporter strain. In 26 bladder tumors treated with BCG instillations there was a statistical difference (p = 0.0075) in the response to BCG therapy between 18 tumors with and 8 without alterations using functional assays in yeast. CONCLUSIONS: The p53 mutations, using functional assay in yeast, inactivate the transcription of p21 and Bax genes, and based on these preliminary results could have a useful predictive value for BCG therapy response in bladder cancer. PMID- 10379743 TI - The value of a second transurethral resection in evaluating patients with bladder tumors. AB - PURPOSE: The role of a routine second transurethral resection in evaluating and managing bladder tumors is defined. MATERIALS AND METHODS: From January to October 1998, 150 patients with new or recurrent bladder tumors underwent repeat transurethral resection within 2 to 6 weeks after the initial resection, and the results, including the presence of residual tumor and tumor stage, were compared. RESULTS: Of the 150 cases 36 (24%) had no and 114 (76%) had residual tumor on repeat transurethral resection. Of 96 cases with superficial (Ta, Tis, T1) bladder tumors 72 (75%) had residual noninvasive tumor and 28 (29%) were up staged to invasive tumor. Among 54 patients with a muscle invasive tumor 12 (22%) had no residual tumor on repeat transurethral resection. Results of the second resection changed tumor treatment in 50 patients (33%). CONCLUSIONS: Many patients with bladder tumors have tumor present after an initial trans-urethral resection. Routine repeat resection is advised to control noninvasive tumors and to detect residual tumor invasion. PMID- 10379744 TI - Quality of life after radical cystectomy for bladder cancer in patients with an ileal conduit, cutaneous or urethral kock pouch. AB - PURPOSE: Radical cystectomy for bladder cancer is associated with many changes in bodily function with sexual and urinary dysfunction most prevalent. However, little research has been done on how efforts to improve erectile function relate to quality of life. Also, the psychological benefits associated with continent urinary diversion have not been fully explored. We compared long-term quality of life outcomes among 3 urinary diversion groups, and between patients who had and had not received an inflatable penile prosthesis. MATERIALS AND METHODS: The 224 participating patients completed 4 self-reporting questionnaires, including the profile of mood states, and adapted versions of the sexual history form, body image dissatisfaction scale and quality of life questionnaire. We compared self reports of emotional distress, global quality of life, sexuality, body image dissatisfaction, urinary diversion problems, and problems with social, physical and functional activities in patients with advanced bladder cancer who underwent urinary diversion, including an ileal conduit in 25, cutaneous Kock pouch in 93 and urethral Kock pouch in 103. Patients who had or had not received an inflatable penile prosthesis after cystectomy were also compared in regard to quality of life variables. RESULTS: Regardless of type of urinary diversion the majority of patients reported good overall quality of life, little emotional distress and few problems with social, physical or functional activities. Problems with urinary diversion and sexual functioning were identified as most common. After controlling for age analysis of variance showed no significant differences among urinary diversion subgroups in any quality of life area. However, t tests controlling for age indicated that penile prosthesis placement was significantly associated with better sexual function and satisfaction. CONCLUSIONS: Quality of life appears good in these long-term survivors of advanced bladder cancer. The type of urinary diversion does not appear to be associated with differential quality of life. Findings suggest that physicians may wish to discuss urinary diversion problems and sexual dysfunction as long term correlates of radical cystectomy for bladder cancer. Furthermore, they may also wish to discuss the option of erectile aids in men with erectile dysfunction after cystectomy. PMID- 10379745 TI - Urinary incontinence and depression. AB - PURPOSE: Serotonergic neuronal systems have been implicated in anxiety and depression. Because descending serotonin pathways from the brain stem inhibit bladder contractions, we postulated that depression associated with altered serotonin function may predispose to urge incontinence. We demonstrate an association between depression and idiopathic urge incontinence. MATERIALS AND METHODS: A total of 115 consecutive incontinent patients presenting to an incontinence clinic were compared to 80 continent controls. Patients were queried for a history of depression and completed a Beck Depression Inventory (BDI). Cases were classified by history and video urodynamics as genuine stress (36), urge (44) or mixed (35) incontinence. RESULTS: A BDI of greater than 12 and/or a history of depression was noted in 30% of incontinent patients and 17% of controls (odds ratio 2.3, 95% confidence interval 1.0 to 5.0, p = 0.044). An abnormal BDI or history of depression was revealed in 60% of patients with idiopathic urge incontinence (p<0.001). Patients with stress or urge incontinence due to neuropathology or obstruction had no greater odds of having depression than continent controls. CONCLUSIONS: These data suggest a strong association between depression and idiopathic urinary incontinence. This link may be due to altered serotonin function and may help explain the efficacy of serotonergic based antidepressants in the treatment of urge incontinence. PMID- 10379747 TI - Serum and semen prostate specific antigen concentrations are different in young spinal cord injured men compared to normal controls. AB - PURPOSE: Recent investigations have indicated that factors within the seminal plasma may contribute to the condition of low sperm motility in men with spinal cord injury. To determine whether the prostate gland functions normally in these men we chose prostate specific antigen (PSA) as a marker of prostatic function, and compared serum and semen concentrations in spinal cord injured and healthy noninjured men. MATERIALS AND METHODS: The study included 21 spinal cord injured men (mean age 33.3+/-1.2 years) and 22 noninjured normal men (mean age 30.3+/-1.5 years). Blood was obtained from subjects following at least 24 hours of abstinence from ejaculation and serum PSA was determined by modified enzyme immunoassay. Antegrade ejaculates from all subjects were frozen to -80 C, exactly 15 minutes after collection. Seminal plasma PSA was determined using Hybritech Tandem MP assay. RESULTS: Mean serum PSA concentration was 1.20+/-0.19 ng./ml. in spinal cord injured and 0.69+/-0.07 ng./ml. in noninjured men (p<0.02). Mean seminal plasma PSA concentration was 0.59+/-0.11 mg./ml. in spinal cord injured and 1.29+/-0.15 mg./ml. in noninjured men (p<0.001). CONCLUSIONS: Our findings of elevated serum and decreased seminal plasma PSA concentrations indicate that prostatic secretory dysfunction is present in men with spinal cord injury. PMID- 10379746 TI - Comparison of bilateral versus unilateral varicocelectomy in men with palpable bilateral varicoceles. AB - PURPOSE: The left varicocele is usually larger in men with bilateral varicoceles. We hypothesized that most of the benefit of varicocelectomy would derive from repair of the larger varicocele. To test this hypothesis we prospectively compared the effect of unilateral versus bilateral microsurgical varicocelectomy in men with large (grade III) or moderate (II) left varicocele associated with small but palpable (I) right varicocele. MATERIALS AND METHODS: A total of 91 patients were prospectively followed and included in the study. Of the patients 65 underwent bilateral and 26 underwent unilateral left repair. All patients underwent preoperative and postoperative semen analysis. RESULTS: Motile sperm concentration increased from 12.1+/-1.7 to 23.7+/-31.8 (95.8% change) in the bilateral group compared with an increase from 19.5+/-21.4 to 27.8+/-34.8 (42.6% change) in the unilateral group (p<0.05). Similarly, sperm concentration increased from 23.8+/-29.5 to 48.6+/-61.3 (157.6% change) in the bilateral group compared with an increase from 41.1+/-40.9 to 59.5+/-66.7 (44.8% change) in the unilateral group (p<0.05). CONCLUSIONS: Bilateral varicocelectomy resulted in significantly greater improvement in post-operative seminal parameters than unilateral repair in patients with grades II to III left varicocele associated with grade I right varicocele. Even a small, unrepaired palpable right varicocele continues to have a detrimental effect on bilateral testis function. Men with bilateral palpable varicoceles require bilateral repair. PMID- 10379748 TI - The effects of transurethral needle ablation and resection of the prostate on pressure flow urodynamic parameters: analysis of the United States randomized study. AB - PURPOSE: We evaluated the effects of transurethral needle ablation and prostate resection on pressure flow urodynamic parameters in men with benign prostatic hyperplasia (BPH), compared symptomatic and objective parameters of efficacy 6 months after initial treatment, and determined whether urodynamic assessment may predict symptomatic improvement. MATERIALS AND METHODS: We enrolled 121 patients with clinical BPH, American Urological Association symptom index of 13 or greater and maximum urinary flow of 12 ml. per second or less in a randomized study comparing transurethral needle ablation to prostate resection at 7 institutions in the United States. Patients underwent baseline and followup assessments at 6 months, including pressure flow studies. RESULTS: Patients who underwent each procedure had statistically and clinically significant improvement in symptom index, BPH impact index and quality of life score. After needle ablation and prostate resection maximum flow improved from 8.8 to 13.5 (p<0.0001) and 8.8 to 20.8 ml. per second (p<0.0001), detrusor pressure at maximum flow decreased from 78.7 to 64.5 (p = 0.036) and 75.8 to 54.9 cm. water (p<0.001), and the Abrams Griffiths number decreased from 61.2 to 37.2 (p<0.001) and 58.3 to 10.9 (p<0.001), respectively. At 6 months the differences in transurethral needle ablation and prostate resection were significant in terms of maximum flow (p<0.001) and the Abrams-Griffiths number (p<0.001) but not detrusor pressure at maximum flow or symptom assessment tools. The presence or absence of urinary obstruction at baseline did not predict the degree of symptomatic improvement in either treatment group. CONCLUSIONS: Transurethral needle ablation and prostate resection induce statistically and clinically significant improvement in various quantitative symptom assessment questionnaires at 6 months. The parameters of free flow rates and invasive pressure flow studies also significantly improve after each treatment. However, transurethral prostate resection induces a significantly greater decrease in the parameters of obstruction. Baseline urodynamic parameters do not predict the degree of symptomatic improvement and they may not be helpful in patient selection for transurethral needle ablation. PMID- 10379749 TI - Pilot evaluation of venlafaxine for the treatment of hot flashes in men undergoing androgen ablation therapy for prostate cancer. AB - PURPOSE: Hot flashes may be a significant clinical problem in men undergoing androgen deprivation therapy with gonadotropin releasing hormone analogues, oral antiandrogens and/or surgical bilateral orchiectomy. Anecdotal information suggests that a low dose of the relatively new antidepressant venlafaxine may abrogate this clinical problem. We developed the current pilot trial to investigate further whether venlafaxine alleviates hot flashes in such men. MATERIALS AND METHODS: The study included men in whom substantial hot flashes were associated with androgen deprivation therapy. Hot flash data were collected by daily diary questionnaires during a 1-week baseline period when no therapy was given for hot flashes, as well as for the next 4 weeks when study participants received 12.5 mg. venlafaxine orally twice daily. Questionnaires completed during the 4 weeks ofvenlafaxine therapy also documented data on potential drug toxicity. RESULTS: Of the 16 evaluable patients who completed the study 10 (63%) had a greater than 50% decrease in hot flash score, as determined using the formula, frequency x severity, by week 4 of treatment versus the baseline week. Median weekly hot flash scores decreased 54% from baseline during week 4 of venlafaxine therapy. Average incidence of severe and very severe hot flashes was reduced from 2.3 daily at baseline to 0.6 daily at study end (p = 0.003). Therapy was generally well tolerated. CONCLUSIONS: Venlafaxine hydrochloride appears to represent an efficacious new method for alleviating hot flashes in men undergoing androgen ablation therapy. Further evaluation of this compound for alleviating hot flashes is indicated. PMID- 10379750 TI - Can perineural invasion on prostate needle biopsy predict prostate specific antigen recurrence after radical prostatectomy? AB - PURPOSE: We evaluated the role of perineural invasion identified on prostate needle biopsy as a predictor of prostate specific antigen (PSA) recurrence after radical prostatectomy. MATERIALS AND METHODS: Between 1993 and 1998 radical prostatectomy was performed in 319 consecutive patients. Prostate needle biopsies were reviewed in all cases. We compared perineural invasion with other preoperative parameters, including digital rectal examination, PSA and biopsy Gleason score, for the ability to predict PSA recurrence with recurrence defined as any serum PSA level greater than 0.2 ng./ml. RESULTS: Perineural invasion was identified on 77 of 319 preoperative prostate biopsies (24%). There was PSA recurrence in 46 patients (14.4%) at a mean followup of 25.4 months (range 0.2 to 62.1). Perineural invasion statistically correlated with PSA recurrence. Kaplan Meier analysis revealed disease-free survival rates of 24 versus 64% when perineural invasion was and was not present in the prostate biopsy (p = 0.0003, log rank 12.92). Multivariate analysis demonstrated that perineural invasion (p = 0.012) and PSA (p = 0.005) were independent preoperative predictive factors of PSA recurrence. When perineural invasion was compared with postoperative parameters, including disease stage, surgical margins and seminal vesicle invasion, it was not an independent predictor because it closely correlated with tumor stage. CONCLUSIONS: Perineural invasion on preoperative prostate needle biopsy is a strong independent predictor of PSA recurrence in patients in whom prostate cancer was treated with radical prostatectomy. PMID- 10379751 TI - Have complication rates decreased after treatment for localized prostate cancer? AB - PURPOSE: The American Urological Association Prostate Cancer Clinical Guidelines Panel reviewed 12,501 publications on prostate cancer from 1955 to 1992 to determine whether the complication rates of external beam radiation therapy, interstitial radiotherapy and radical prostatectomy have decreased. MATERIALS AND METHODS: Complications reported in at least 6 series, study duration and sample sizes were extracted. Year specific study weighted mean patient ages and complication rates were computed. Regression analysis was performed of the study year on weighted mean patient age and complication rate. RESULTS: Study year had a significant effect on mean patient age and rate of the majority of complications examined. Data indicated a gradual increase in study patient age and a simultaneous decrease in complications from 1960 to 1990. CONCLUSIONS: Complication rates in the treatment of localized prostate cancer have decreased during the last 20 to 40 years. This decrease occurred despite evidence that the average age of treated patients had increased during the same period. PMID- 10379752 TI - Correlates of dissatisfaction with treatment in patients with prostate cancer diagnosed through screening. AB - PURPOSE: We evaluated correlates of patient reported dissatisfaction with treatment of prostate cancer detected by screening. MATERIALS AND METHODS: We performed a cross-sectional retrospective study to evaluate the correlates of dissatisfaction with treatment in 1,651 patients in whom prostate cancer was detected through serial screening. We included demographic and clinical characteristics in the independent and control variables, and we validated measurements of quality of life outcomes. RESULTS: Overall 11% of patients were dissatisfied with the treatment received. Differences in the rates of dissatisfaction with treatment were not statistically significant across treatment groups (11% for retropubic radical prostatectomy, 21% for perineal radical prostatectomy, 14% for radiotherapy, 8% for observation, 8% for hormonal treatment and 4% for cryoablation, p = 0.1). Patient age, race, followup interval, marital status, education and co-morbid conditions were not significant correlates of dissatisfaction with treatment (for all characteristics p> or =0.05). Urinary function and bothersomeness were associated with dissatisfaction with treatment (p<0.0001), whereas sexual function and bothersomeness were not (p>0.05). Multivariate analysis revealed that urinary function and bothersomeness were also the only significant correlates of dissatisfaction with treatment. CONCLUSIONS: Of patients in whom prostate cancer was detected by screening 11% were dissatisfied with treatment. Urinary function and bothersomeness were the only important correlates of dissatisfaction. PMID- 10379753 TI - The value of comparative volumetric analysis of urinary and blood erythrocytes to localize the source of hematuria. AB - PURPOSE: We evaluate comparative volumetric analysis of blood and urinary red blood cells (RBCs) to identify the source of hematuria. Comparative volumetric analysis is defined as the difference between mean corpuscular erythrocyte volume in peripheral blood (MCVB) diluted in urine supernatant after centrifugation and mean corpuscular volume of urinary erythrocytes (MCVU). The potential of MCVB MCVU to distinguish the origin of hematuria is compared to MCVU alone. The fundamental hypothesis is that RBCs that can go through the glomerulus will be smaller than those from the collecting system or lower urinary tract, thus having a smaller MCVU and larger difference between MCVB and MCVU. MATERIALS AND METHODS: A prospective detailed urological evaluation was performed on 210 patients with glomerular or nonglomerular hematuria detected by urinary sediment, clinical radiological evaluation, endoscopy, cytology and sometimes bladder or renal biopsy. After evaluation 24 cases with an uncertain source of hematuria were excluded from study. Specialized urinalysis, volumetric analysis and clinical investigation were performed in a blind fashion. MCVU and MCVB-MCVU were registered for every patient. The Technicon H-3 system with angle laser scattering dual system allowed measurement of mean corpuscular volume in a minimal number of RBCs, and resuspension of RBC pellets in the same urinary supinate avoided effects of osmolarity and pH on RBC size and shape. Reproducibility in assessing the index was tested in 50 cases in which comparative volumetric analysis was repeated on 2 consecutive days. Unpaired t test was performed, and a threshold value of MCVB-MCVU with maximum sensitivity and specificity to detect glomerular hematuria was identified. The potential of urinary and comparative volumetric analysis to distinguish the source of hematuria was evaluated and compared by receiver operating characteristics curve analysis. RESULTS: Hematuria was nonglomerular in 53 (28.4%) and glomerular in 133 (71.6%) patients. Mean MCVB-MCVU was significantly different for nonglomerular (0.6 fl.) and glomerular (30.5 fl.) sources (p<0.0001). There was a correlation between repeat independent measures of MCVU and MCVB-MCVU. The highest positive predictive value to detect a glomerular origin is desirable so that unnecessary investigation can be obviated without the risk of missing a nonglomerular source. With a limit of 16 fl. specificity and positive predictive value were 98 and 99%, respectively. Receiver operating characteristics curve analysis to localize the source of hematuria revealed significant differences in favor of comparative volumetric analysis versus urinary volumetric analysis alone. CONCLUSIONS: MCVB-MCVU using the Technicon H-3 system is a useful noninvasive and accurate method to locate the source of hematuria. A value of 16 fl. or greater practically rules out a nonglomerular origin and obviates further urological investigation. We have incorporated this investigation in our diagnostic algorithm for hematuria. PMID- 10379754 TI - Prepubic urethrectomy with urethral stripping. AB - PURPOSE: Prepubic urethrectomy is a simple, safe alternative to perineal urethrectomy. The lithotomy position can be avoided and, thus, operative time and risk of deep venous thrombosis are decreased. We developed a simple modification because of difficulty in dissecting the bulbous urethra. MATERIALS AND METHODS: From 1996 through 1998 prepubic urethrectomy was performed using a modified procedure in 21 patients with invasive bladder carcinoma undergoing radical cystectomy and supravesical diversion. After periurethral mobilization the urethra was cannulated with an 18F catheter, sutured distal and stripped free. RESULTS: Operative time decreased to 20 to 30 minutes with no significant postoperative complications. CONCLUSIONS: Our modification of prepubic urethrectomy is safe, fast and easy. PMID- 10379755 TI - Activation of the kallikrein kinin system in interstitial cystitis. AB - PURPOSE: We investigated whether the kallikrein kinin system is activated in interstitial cystitis by measuring urinary excretion rates of kinin peptides, active and total kallikrein, and the kininase neutral endopeptidase in women with interstitial cystitis. We compared these excretion rates to a control group of women with stress incontinence and normal bladder function. MATERIALS AND METHODS: Catheter urine was collected from subjects during a water diuresis (approximately 10 ml. per minute) before and after distention of the bladder with 100 ml. water. The contribution of the bladder wall to urinary kinins was assessed by measuring the change in kinin levels after 2 minutes of bladder stasis before and after distention. RESULTS: Absolute bradykinin and kallidin excretion rates were similar in women with interstitial cystitis and control subjects. Two minutes of bladder stasis after bladder distention increased urinary bradykinin (p = 0.02) but not kallidin excretion rates. Active and total kallikrein excretion rates were similar in patients with interstitial cystitis and control subjects. Neutral endopeptidase excretion rates were reduced in the initial urine collection from subjects with interstitial cystitis but were similar in both groups during later collection periods. CONCLUSIONS: These data provide evidence for increased bradykinin levels in the bladder wall of subjects with interstitial cystitis, which may be due in part to reduced neutral endopeptidase levels. These increased bradykinin levels may participate in the pathogenesis and symptomatology of interstitial cystitis. PMID- 10379756 TI - An assessment of the surgical outcome and urodynamic effects of the pubovaginal sling for stress incontinence and the associated urge syndrome. AB - PURPOSE: We assessed the urodynamic changes after pubovaginal sling procedure for stress incontinence, particularly in regard to the associated symptoms of urgency, frequency, nocturia and urge incontinence, known as the urge syndrome. MATERIALS AND METHODS: A total of 85 women with proved stress incontinence underwent a pubovaginal sling procedure using rectus fascia between 1992 and August 1996. Of the women 41 (48%) had undergone previous anti-incontinence surgery and 59 (69%) had the associated urge syndrome. There was at least some degree of hypermobility in 51 cases and type III stress incontinence was diagnosed in 34. Patients were assessed with a questionnaire and video urodynamics preoperatively and 3 months postoperatively. Preoperative and postoperative ambulatory studies were performed in 25 cases. RESULTS: Of the 85 patients 83 (97%) were symptomatically cured of stress incontinence. The urge syndrome resolved in 32 patients (69%), almost all of whom had a closed bladder neck at rest. Overall bladder neck incompetence at rest decreased from 57 to 18% (p<0.001). Of 27 patients with the persistent urge syndrome postoperatively 9 (41%) had an open bladder neck at rest compared to 4 of 50 (8%) without urge incontinence (p<0.01). Despite symptomatic control of stress incontinence in 83 patients (97%), only 66 were satisfied with the surgical result, mainly because of the persistent urge syndrome in 27. Despite care to avoid obstruction overall, there were statistically significant obstructive changes in detrusor pressure at maximum flow rate, maximum flow rate and residual urine volumes. CONCLUSIONS: The pubovaginal sling is effective in curing genuine stress incontinence and, when correctly placed at the right tension, the associated urge syndrome also can be managed, usually by achieving bladder neck closure at rest. However, despite careful maneuvers, obstruction occasionally persists. PMID- 10379757 TI - Long-term results of colpocystourethropexy for persistent or recurrent stress urinary incontinence. AB - PURPOSE: We review the long-term outcome of colpocystourethropexy for persistent or recurrent stress urinary incontinence after suspension procedure failure. MATERIALS AND METHODS: Medical records and preoperative studies were reviewed of 60 patients (mean age 60.8 years) who had undergone colpocystourethropexy after at least 1 suspension procedure (range 1 to 8, mean 2.7). Patient responses to a standardized questionnaire regarding overall health, degree of satisfaction with colpocystourethropexy, presence or absence of leakage, and pattern and degree of leakage were elicited by telephone or mail and compared with preoperative status. Results were graded according to the degree of satisfaction and number of pads used daily. Patients with persistent incontinence were reevaluated with video urodynamic studies. RESULTS: Mean interval since colpocystourethropexy was 6.9 years. Successful results (greater than 80% satisfaction and the use of 1 or no pad daily) were reported by 41 patients (69%), who were significantly younger at the time of surgery than those with unsatisfactory results. In the latter group significant urge incontinence was present in 61% before the repair and in 63% postoperatively, suggesting an additional nonanatomical cause, which was confirmed by postoperative video urodynamic studies. CONCLUSIONS: When colpocystourethropexy was used for persistent urinary incontinence after previous surgical repair two-thirds of the patients had excellent long-term results. In patients with less satisfactory results a nonanatomical cause of urinary incontinence was a major factor. PMID- 10379758 TI - Urodynamic characterization of nonobstructive voiding dysfunction in symptomatic elderly men. AB - PURPOSE: The pathogenesis of lower urinary tract symptoms in men without bladder outlet obstruction has not been well characterized. Therefore, we defined the urodynamic abnormalities associated with symptomatic nonobstructive voiding dysfunction, and determined the relationship between age and type of dysfunction. MATERIALS AND METHODS: Video urodynamic studies of symptomatic men without outlet obstruction were examined. The criterion for a normal bladder outlet was a pressure gradient across the prostatic urethra of 5 cm. water or less in the absence of distal stricture. A maximum isometric contraction pressure less than 60 cm. water was regarded as impaired detrusor contractility. Detrusor instability was defined as involuntary detrusor contractions during filling or the inability to suppress a detrusor contraction after initiation of flow. Patients were categorized into 4 groups based on the urodynamic findings. RESULTS: Of 193 men (mean age 69.6+/-10.5 years) 40.9% had detrusor instability (group 1), 31.1% had impaired contractility (group 2), 10.8% had detrusor instability and impaired contractility (group 3), and 17.1% were urodynamically normal (group 4). Average patient age was significantly lower in group 4 than all other groups. Bladder capacity was lowest in group 1, and group 3 had the lowest voiding efficiency. Maximum flow rate, bladder compliance and symptom scores were not different among the 4 groups. The prevalence of detrusor instability with and without impaired contractility increased, while the proportion of patients without urodynamic abnormalities decreased with age. Bladder contractility did not correlate with age. CONCLUSIONS: The nonobstructed patient population comprises several groups that are functionally distinct while symptomatically similar. Thus, treatment of nonobstructed cases based on symptoms may lead to inappropriate pharmacological therapy and unsuccessful clinical outcomes. PMID- 10379759 TI - Evaluation and therapeutic approaches of voiding and erectile dysfunction in neurological Behcet's syndrome. AB - PURPOSE: Behcet's syndrome is a progressive inflammatory disease which involves multiple systems. It is characterized by 3 main symptoms of iridocyclitis, and oral and genital ulcerations. Nervous system involvement is seen rarely in this clinical entity and is known as neurological Behcet's syndrome. Inflammation usually occurs in the brain stem, cerebellum and medulla spinalis. Voiding and erectile dysfunction can be due to progressive inflammatory reactions in the nervous and vascular systems. We prospectively evaluated the dysfunctional bladder and penis, and therapeutic options were evaluated prospectively. MATERIALS AND METHODS: A total of 24 consecutive patients diagnosed with neurological Behcet's syndrome after neurological evaluation were enrolled in this study. Neurological involvement and localization of the nervous system were proved on evaluation. Voiding and erectile dysfunction was evaluated regardless of the presence of related symptoms, and the results were compared with those of controls. Patients with voiding dysfunction on urodynamic study were treated and reevaluated symptomatically after 3 and urodynamically after 6 months. RESULTS: The rate of erectile dysfunction in neurological Behcet's syndrome was 63%. Mixed type vasculogenic impotence, arterial insufficiency, veno-occlusive dysfunction and neurogenic impotence were identified in 7, 2, 2 and 1 patient, respectively. Detrusor instability was demonstrated in 12 patients with urgency incontinence, including 3 with detrusor-sphincter dyssynergia. Brain stem localization was determined in these patients on neurogenic evaluation. Significant improvement was observed with anticholinergic treatment and clean intermittent catheterization in 3 patients with detrusor-sphincter dyssynergia. Hypersensitive and hypocompliant detrusor was noted in patients with neurological Behcet's syndrome who had normal voiding habits. CONCLUSIONS: Incontinence or irritable bladder symptoms should not be considered innocuous clinical findings in neurological Behcet's syndrome. Lower urinary tract function should be evaluated in all patients with this neurological syndrome. The incidence of erectile dysfunction is approximately 65% and the therapeutic approach should be determined according to lower urinary tract function. PMID- 10379760 TI - Myasthenia gravis as a paraneoplastic syndrome associated with renal cell carcinoma. PMID- 10379761 TI - Diagnosis and treatment of leptomeningeal metastases in a patient with renal carcinoma responding to 5-fluorouracil and gemcitabine. PMID- 10379762 TI - Stomal recurrence of bladder carcinoma after cystectomy. PMID- 10379763 TI - Extensive bladder and urethral calculi detected with computerized tomography: diagnosis and management. PMID- 10379764 TI - Primary extraskeletal Ewing's sarcoma of the external genitalia. PMID- 10379765 TI - Mixed germ cell testis tumor in an 86-year-old man. PMID- 10379766 TI - Mixed germ cell tumor of the testicle presenting with a sacrococcygeal mass and no evidence of retroperitoneal adenopathy. PMID- 10379767 TI - Urethral obstruction due to protruding prostatic calculi. PMID- 10379768 TI - Malignant peripheral nerve sheath tumor of the prostate: a rare manifestion of neurofibromatosis type 1. PMID- 10379769 TI - Re: Conservative surgical management of bilateral Wilms tumor: results of the United Kingdom Children's Cancer Study Group. PMID- 10379770 TI - Re: An effective technique to facilitate radiographic stone visualization with an internal stent during shock wave lithotripsy. PMID- 10379771 TI - Re: Letter to the editor re: editorial: alternatives to appendix in construction of a Mitrofanoff stoma. PMID- 10379772 TI - Re: Development of noninvasive velocity flow video urodynamics using Doppler sonography. Part I: Experimental urethra and development of noninvasive velocity flow video urodynamics using Doppler sonography. Part II: Clinical application in bladder outlet obstruction. PMID- 10379773 TI - Re: Effect of cardiopulmonary bypass on urethral blood flow as measured by laser Doppler flowmetry. PMID- 10379774 TI - Re: Disappointing initial results with transurethral alprostadil for erectile dysfunction in a urology practice setting. PMID- 10379775 TI - Re: Benefits of laparoscopy and the Jones technique for the nonpalpable testis. PMID- 10379776 TI - Re: Clinical stage I testis cancer: long-term outcome of patients on surveillance. PMID- 10379777 TI - Re: Time to normalization of serum testosterone after 3-month luteinizing hormone releasing hormone agonist administered in the neoadjuvant setting: implications for dosing schedule and neoadjuvant study consideration. PMID- 10379778 TI - Re: Time to normalization of serum testosterone after 3-month luteinizing hormone releasing hormone agonist administered in the neoadjuvant setting: implications for dosing schedule and neoadjuvant study consideration. PMID- 10379779 TI - Re: Prostatic infarction/infection in acute urinary retention secondary to benign prostatic hyperplasia. PMID- 10379780 TI - Re: Transient lower extremity neurapraxia associated with radical perineal prostatectomy: a complication of the exaggerated lithotomy position. PMID- 10379781 TI - Re: editorial: Urological diseases of women, and women in urology. PMID- 10379782 TI - Antegrade pyelography before pyeloplasty via dorsal lumbar incision. AB - PURPOSE: The need for contrast imaging of the ureter before routine pediatric pyeloplasty is controversial. We evaluated the use of antegrade pyelography for upper tract imaging before pyeloplasty via dorsal lumbar incision. MATERIALS AND METHODS: The records of all patients who underwent pyeloplasty from April 1994 through April 1998 at our institution were reviewed. The findings and outcome of patients with presumed ureteropelvic junction obstruction in whom antegrade pyelography was performed under the same anesthetic were assessed, and those in whom this procedure changed the planned operative approach were identified. RESULTS: Antegrade pyelography was performed without complication in 72 patients before planned pyeloplasty and 2 attempts were unsuccessful. In 10 cases (14%) plans for dorsal lumbar incision were abandoned based on findings of renal malrotation in 3, ureteral stricture in 2, ureterovesical junction obstruction in 2, unusually low or high position of the ureteropelvic junction in 1 each, and concurrent ureteropelvic and ureterovesical junction obstruction in 1. The study was misinterpreted in 1 case of renal malrotation and 1 case of horseshoe kidney, and the dorsal approach was used. In 1 of these cases conversion to an anterior approach was required. A nonobstructing ureterovesical junction was seen in 2 other patients who had ureteropelvic junction obstruction with mild ureteral dilatation on ultrasound. CONCLUSIONS: The dorsal lumbar incision may provide inadequate exposure in certain patients with upper tract obstruction. Antegrade pyelography is a simple, safe and useful technique to visualize the collecting system before planned pyeloplasty via dorsal lumbar incision, allowing the surgeon to choose a more suitable operative approach or procedure when warranted. PMID- 10379783 TI - Absence of antisperm surface antibodies in prepubertal boys with cryptorchidism and other anomalies of the inguinoscrotal region before and after surgery. AB - PURPOSE: Although the prepubertal immune system cannot recognize postmeiotic germ cell antigens, an overall 21 to 28% incidence of antisperm antibodies directed at these antigens has been reported preoperatively in prepubertal children with cryptorchidism and other inguinoscrotal anomalies. We investigated the prevalence of antisperm antibodies in these prepubertal patients before and after surgery. MATERIALS AND METHODS: We examined 82 prepubertal boys 0.6 to 13.2 years old, including 33 with unilateral cryptorchidism, 21 with inguinoscrotal anomalies and 28 who were normal. IgG, IgM and IgA antisperm antibodies were determined by the indirect Immunobead test. Serum testing was repeated 1 and 2 years postoperatively and annually for 2 more years in the normal children. Also sera from 183 infertile men 21 to 47 years old with a history of cryptorchidism and/or inguinal hernia operated on in childhood were similarly studied. RESULTS: Of the adults 70 (39%) tested IgG positive, including 12 (7%) who were also IgA positive, and all tested IgM negative. Repeat measurements were negative for all IgG, IgA and IgM isotypes in all children, patients and controls. CONCLUSIONS: We conclude that there are no antibodies to sperm surface antigens in prepubertal children with cryptorchidism and inguinoscrotal anomalies before and within 2 years after surgery. Autoimmunity against postmeiotic sperm membrane antigens is apparent in adults only. PMID- 10379784 TI - Functional, social and psychosexual adjustment after vaginal reconstruction. AB - PURPOSE: We assessed the long-term functional, social and psychosexual outcome in children who underwent vaginal reconstruction. MATERIALS AND METHODS: We interviewed and assessed the psychological development of 16 women 17 to 28 years old (mean age 22) who underwent vaginal reconstruction only or in combination with other urogenital reconstructive procedures at ages 11 months to 18 years (mean 8.8). Psychological measures included the Beck Depression Inventory, Draw-a Person test and Linkowski acceptance of disability scale as well as a standard questionnaire evaluating the sexual adjustment, social adjustment and ability for self-support of these women. RESULTS: Mean Beck Depression Inventory was 8.5 with less than 9 defined as minimal depression. Mean acceptance of disability score was 83.9 (range 54 to 94), indicating that patients were well adjusted with respect to the disability. Functionally 10 women were satisfied with the appearance of the vagina, 4 were neutral and 2 were dissatisfied. A total of 12 patients had no doubts about their female identity but 2 had occasional, 1 had significant and 1 had chronic doubts. Of the 16 patients 12 have completed high school, 3 are still in high school and 1 has withdrawn from high school. Of the 12 women who have completed high school 9 are currently in college and 3 have completed college. Socially all 16 participants rated family relationships as good and 13 were at least satisfied with their social life. Of the 16 women 12 have had a sexual encounter, including 1 who did not achieve orgasm. Six women are involved in long-term relationships, of whom 1 is married. In regard to the future all patients believe that they will be independent and financially stable with a fulfilling career. CONCLUSIONS: While women who have undergone vaginal reconstruction may be at risk for avoiding interpersonal relationships and sexual intimacy, we did not note this finding in our series. The majority of these patients were well adjusted to their physical condition, and had a high level of education and a stable family life. PMID- 10379785 TI - Functional, social and psychosexual adjustment after vaginal reconstruction. PMID- 10379786 TI - Hair tourniquet syndrome of the clitoris. PMID- 10379787 TI - Review article: the role of tumor necrosis factor in renal ischemia-reperfusion injury. AB - Renal ischemia-reperfusion injury induces a cascade of events leading to cellular damage and organ dysfunction. Tumor necrosis factor-alpha (TNF), a potent proinflammatory cytokine, is released from the kidney in response to, and has been implicated in the pathogenesis of, renal ischemia-reperfusion injury. TNF induces glomerular fibrin deposition, cellular infiltration and vasoconstriction, leading to a reduction in glomerular filtration rate (GFR). The signaling cascade through which renal ischemia-reperfusion induces TNF production is beginning to be elucidated. Oxidants released following reperfusion activate p38 mitogen activated protein kinase (p38 MAP kinase) and the TNF transcription factor, NFkappaB, leading to subsequent TNF synthesis. In a positive feedback, proinflammatory fashion, binding of TNF to specific TNF membrane receptors can reactivate NFkappaB. This provides a mechanism by which TNF can upregulate its own expression as well as facilitate the expression of other genes pivotal to the inflammatory response. TNF receptor binding can also induce renal cell apoptosis, the major form of cell death associated with renal ischemia-reperfusion injury. Anti-TNF strategies targeting p38 MAP kinase, NFkappaB, and TNF itself are being investigated as methods of attenuating renal ischemic injury. The control of TNF production and activity represents a realistic goal for clinical medicine. PMID- 10379789 TI - Cocaine induced apoptosis in rat testes. AB - PURPOSE: Exposure of rats to chronic cocaine results in disruption of spermatogenesis including reduction of germ cells. However, the cellular mechanism responsible for the testicular damage in testes is still unknown. We have studied the role of apoptosis in cocaine induced testicular damage. MATERIALS AND METHODS: Thirty-day-old male Sprague-Dawley rats were given cocaine hydrochloride (15 mg./kg. body weight) subcutaneously daily for 90 days. Control animals received equal volumes of normal saline daily for 90 days. Testes were removed at 15, 30, 60, and 90 days of cocaine administration. In situ detection of germ cells with DNA strand breaks in paraffin-embedded testicular section (5 microm.) was achieved by the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP in situ nick end-labeling (TUNEL) method. DNA fragmentation was also determined by gel electrophoresis. RESULTS: Apoptotic cells were found in the spermatocytes and spermatogonia of germinal epithelium. Less than 7% of seminiferous tubule cross sections showed a high level of apoptosis (> or =3 apoptotic cells per tubule) in control animals compared with experimental group where 25% of the tubules showed a high level of apoptosis (p<0.05). The number of apoptotic cells was significantly increased by 15 days, peaked at 30 days and persisted up to 90 days of cocaine exposure when compared with controls (p<0.05). DNA isolated from the cocaine treated testes displayed a clear ladder pattern whereas the DNA from controls did not. CONCLUSIONS: The experimental results presented here suggest that cocaine exposure leads to significant apoptosis in rat testes and the mechanism of cocaine induced testicular injury may be related to the induction of apoptosis. PMID- 10379788 TI - Urethral afferent nerve activity affects the micturition reflex; implication for the relationship between stress incontinence and detrusor instability. AB - PURPOSE: A causative relationship between stress urinary incontinence (SUI) and detrusor instability has been suspected but never proven. Many women with mixed incontinence have resolution of detrusor instability after surgical correction of SUI. We sought experimental support that stimulation of urethral afferent nerves can induce or change reflex detrusor contractions. MATERIALS AND METHODS: Urethral perfusion pressure and isovolumetric bladder pressure were measured with catheters inserted through the bladder dome in urethane anesthetized female S.D. rats (250 to 300 grams; n = 12). The catheter assembly was seated securely in the bladder neck to block passage of fluid between the bladder and urethra without affecting the nerve supply to the organs. The external urethra was not catheterized. Responses were examined in the control state at a urethral saline perfusion speed of 0.075 ml. per minute. Intraurethral drugs were administered following blockade of striated sphincter activity with intravenous alpha bungarotoxin (0.1 mg./kg.). RESULTS: Stopping the urethral saline infusion caused a significant decrease in micturition frequency in approximately 50% of the animals studied (n = 12). Intraurethral lidocaine (1%) infused at 0.075 ml. per minute caused a slight decrease in urethral perfusion pressure but no change in detrusor contraction amplitude. However, intraurethral lidocaine caused a significant (45%) decrease in the bladder contraction frequency (n = 5). The micturition frequency returned to baseline 30 minutes after stopping lidocaine infusion. Intraurethral infusion of nitric oxide (NO) donors (S-nitroso-N acetylpenicillamine [SNAP] (2 mM) or nitroprusside (1 mM) immediately decreased urethral perfusion pressure by 30 to 37% (n = 5). A 45 to 75% decrease (n = 5) in bladder contraction frequency was also seen, which was similar to that observed following lidocaine. Neither NO donor changed the amplitude of bladder contractions. CONCLUSIONS: These results indicate that in the anesthetized rat activation of urethral afferents by urethral perfusion can modulate the micturition reflex. Thus in patients with stress urinary incontinence, leakage of urine into the proximal urethra may stimulate urethral afferents and facilitate voiding reflexes. This implies that stress incontinence can induce and/or increase detrusor instability. These findings have significant implications for the treatment of patients with mixed urge and stress incontinence. Correction of stress incontinence by surgery or pelvic floor exercise in patients with mixed incontinence may resolve the detrusor instability. PMID- 10379790 TI - Intrinsic drug resistance in primary and metastatic renal cell carcinoma. AB - Much remains to be learned about drug resistance in the biology of RCC and its metastases. We measured MDR-1/P-glycoprotein expression in 19 tumor samples from patients with metastatic RCC by RNase protection and quantitative PCR assays. The median level of the 16 tumor metastases was 4.9 (range: 0.10 to 156.2) relative to the level of 10 assigned to a reference cell line, SW620, which has been characterized as expressing a minimum level of MDR-1. Since these levels were lower than expected for RCC, we asked whether the metastases possessed a phenotype different from primary RCC and examined MDR-1 expression in 5 paired cell lines derived from primary and metastatic RCC. In 8/10 lines, MDR-1 expression was >10. Relative to the level in the primary line, MDR-1 expression was decreased (3 to 50-fold) in 3 metastatic lines, was increased in 1, and unchanged in 1. MRP mRNA expression was lower in the metastatic lines while EGFR expression was variable. IC50 values for 6 compounds (including 4 standard agents and one new Phase 1 agent) were determined for the paired lines. Rhodamine and calcein efflux assays were performed as measures of P-glycoprotein and MRP function. Rhodamine efflux correlated with MDR-1 mRNA expression (r = 0.87) and with the IC50s (r = 0.60) for paclitaxel in the paired cell lines. In contrast, calcein efflux did not correlate with MRP expression. Lastly, MDR-1 expression correlated with cytokeratin 8 (CK8) protein levels, a measure of cellular differentiation. In sum, these data suggest renal cell carcinoma (RCC) metastases have altered MDR-1 expression potentially due to altered differentiation relative to the primary tumor. Thus, the drug resistance phenotype of primary RCC tumors may not reflect that of their metastases. PMID- 10379791 TI - Intravesical bacillus calmette-guerin (BCG) as inducer of tumor-suppressing proteins p53 and p21 Waf1-Cip1 during treatment of superficial bladder cancer. AB - PURPOSE: Previous in vitro investigations recorded an inhibition of cell proliferation by BCG when added to different cell cultures. The induction of apoptosis by BCG is controversial. Our study aimed to evaluate the influence of BCG on the expression of tumor suppressing proteins p53 and p21Waf1-Cip1 and apoptosis of the urothelial cells in vivo. MATERIALS AND METHODS: Twenty-one cases of superficial bladder cancer, treated with TUR and subsequent intravesical BCG, were studied retrospectively. The assays evaluated the expression of p53 and p21Waf1-Cip1 by immunochemistry (IHC), and the presence of apoptosis by TUNEL assay. The estimates were performed, in each case, on the following specimens: one tumor sample and one non-neoplastic sample collected during the TUR which preceded the administration of BCG; one non-neoplastic sample collected 3 months after the diagnosis; and one non-neoplastic sample collected in the first 2 weeks after the completion of the treatment. Samples of 6 cancer recurrences detected during BCG were examined too. RESULTS: As usual for non-neoplastic urothelium, the pre-BCG samples displayed poor p53 and p21Waf1-Cip1 immunoreactivity. By contrast, the samples collected during and in the aftermath of BCG showed an overall increase of the expression of both proteins. The rare occurrence of apoptosis proved to be chronologically unrelated to the BCG treatment. DISCUSSION: The relationship between changes of the IHC features and BCG suggests that BCG, at least under some circumstances, can induce the activation of wild type p53 and p21Waf1-Cip1 in the urothelium. The mechanism of the BCG-p53 status interaction and its role in the antitumor activity of BCG remain to be clarified. PMID- 10379792 TI - Mast cells mediate the severity of experimental cystitis in mice. AB - PURPOSE: We hypothesized that experimental cystitis induced by substance P (SP) or E. coli lipopolysaccharide (LPS) would be less severe in mice rendered mast cell deficient by genetic manipulation. MATERIALS AND METHODS: Two strains of mast-cell deficient mice (WBB6F1- kitW/kitW-v or kitW/kitW-v and WCB6F1-Sl/Sld or Sl/Sld) and their congenic, normal (+/+) counterparts were used. Cystitis was induced in female mice by intravenous injection of SP (0.1 ml.; 10(-6) M) or E. coli LPS (0.1 ml.; 2 mg./ml.), and inflammation was assessed by Evans blue dye extravasation. In a separate group of kitW/kitW-v and congenic normal mice, cystitis was induced by intravesical infusion of SP (0.05 ml.; 10(-5) M) or E. coli LPS (0.05 ml.; 100 microg./ml.) and compared with intravesical pyrogen-free saline (0.05 ml.; 0.9%). Severity of cystitis was determined by histological evaluation of the bladder wall 24 hours after intravesical infusions. RESULTS: Intravenous SP or LPS stimulated increased plasma extravasation in congenic normal mice but not in mast cell-deficient mice. Intravesical SP or LPS resulted in increased edema, leukocytic infiltration, and hemorrhage within the bladder wall in congenic normal mice, but the only histological evidence of inflammation in the bladders of kitW/kitW-v mice was increased hemorrhage in response to LPS. CONCLUSIONS: This study indicates that mast cells modulate the inflammatory response of the bladder to SP and LPS in mice. Although clinical trials of the use of antihistamines to treat or prevent cystitis have not been successful, these results suggest that therapies directed toward preventing mast cell activation may yet prove effective in treating cystitis. PMID- 10379793 TI - Dopaminergic neurotransmission at the paraventricular nucleus of hypothalamus in central regulation of penile erection in the rat. AB - PURPOSE: To investigate whether the paraventricular nucleus of hypothalamus (PVN) is involved in the central regulation of apomorphine-induced penile erection in the rat, and to decipher dopamine receptor subtypes in the PVN that are involved in apomorphine-induced penile erection. MATERIALS AND METHODS: Male adult Sprague Dawley rats (200 to 300 gm.) anesthetized with pentobarbital sodium were used. The intracavernous pressure (ICP), recorded along with systemic and mean arterial pressure (SAP, MAP) as well as heart rate (HR), was measured via a 26-gauge needle inserted into one corpus cavernosum. The PVN was activated by stereotaxically delivered apomorphine hydrochloride (0.1 nmol./100 nl.). Injection of saline into PVN served as a vehicle control. To investigate the participation of dopamine receptor subtypes in the PVN on apomorphine-induced penile erection, D1 or D2 receptor antagonist, SCH-23390 (100 pmol./100 nl.) or sulpiride (100 pmol./100 nl.) respectively, was administered into the PVN prior to subcutaneous application of apomorphine (80 microg./kg.). The effects on ICP of microinjection of D1, D2 or D3 receptor agonist, SKF-38393 (200 pmol./100 nl.), lisuride (200 pmol./100 nl.) or 7-hydroxy-DPAT (200 pmol./100 nl.) respectively, into the PVN were also evaluated. RESULTS: The mean resting ICP was 5.2+/-0.4 mm. Hg. Upon administration of apomorphine into the PVN, there was a significant increase in ICP that peaked at 50.7+/-5.3 mm. Hg and persisted for 45.2+/-18.0 minutes after an onset latency of 677.7+/-311.6 seconds. Yawning and teeth gnashing were also observed in most of animals during the period of ICP increase. There was no significant change in SAP, MAP or HR. In addition, there was no elevation in ICP after administration of saline to the PVN or direct injection of apomorphine into the cavernous tissue. Microinjection of D1 or D2 receptor antagonist into the PVN blocked the increase in ICP after subcutaneous administration ofapomorphine. Direct application of D2, but not D1 or D3 receptor agonist into the PVN, on the other hand, increased the ICP. CONCLUSIONS: Our results demonstrate that application of apomorphine to the paraventricular nucleus of hypothalamus elicited penile erection in the rat. Such an increase in ICP to apomorphine was due mainly to activation of the D2 receptor subtype in the PVN. These observations indicate that PVN may be involved in the central regulation of apomorphine-induced penile erection in the rat. PMID- 10379794 TI - The significance of the difference in bacterial adherence between bladder and ileum using rat ileal augmented bladder. AB - OBJECTIVES: Intestinal segments are frequently used in the reconstruction of the urinary tract. Chronic bacteriuria is frequently observed in these patients, but the reason is not clearly understood. Therefore, we studied the difference in bacterial adherence between bladder and ileum using the rat ileal augmented bladder model to investigate the cause of chronic bacteriuria. MATERIALS AND METHODS: Augmentation of the bladder using ileum and a sham operation were performed under sodium pentobarbital in 102 and 10 Sprague-Dawley rats, respectively. At three months after the operation, urinary pH and plasma concentration of sodium, chloride and potassium were measured and urinary culture was done. Urovirulence factors of Escherichia coli aspirated from augmented bladder were detected by polymerase chain reaction (PCR). Five to six rats with negative urinary cultures after the augmentation were used for each experimental cystitis. E. coli with type I pili aspirated from augmented rats and three clinically isolated strains of E. coli, C5 (type I pili, aerobactin), C92 (type I pili, aerobactin, P fimbriae), and C189 (type I pili, aerobactin, P fimbriae, CNF), were transurethrally inoculated into the augmented bladder of rats. Fourteen days after inoculation, rats were sacrificed and colony-forming units (CFU) per mg. of tissue of bladder and ileum were measured. RESULTS: After operation, urinary pH and the serum level of chloride in all augmented groups were higher than those of the controls. Bacterial colonization was observed in 56 of 89 rats. Most of them were E. coli having only type I pili as a virulence factor. In contrast, the sham operated group revealed no bacterial colonization. In experimental cystitis, E. coli with only type I pili aspirated from augmented rats and E. coli C5 were clearly adhered to ileum rather than to bladder, but E. coli C92 and C189 showed no significant difference with respect to adherence to the two tissues. In experimental cystitis II, E. coli C5 with D-mannose were washed out in 3 of 5 rats by 14 days, while E. coli C5 without D-mannose were not washed out in all rats by 14 days. CONCLUSIONS: These results suggested that the difference in bacterial adherence due to urovirulence factors, especially type I pili, is one of the main causes of asymptomatic bacteriuria after urinary reconstruction. PMID- 10379795 TI - Effect of inflammation on prostatic protein synthesis and luminal secretion in vivo. AB - PURPOSE: Inflammation of the prostate, or prostatitis, can be caused by an infectious process or can occur in a reportedly non-bacterial form, the etiology of which is largely unknown. The present study was undertaken to establish a method of studying prostatic protein synthesis and secretion in vivo and determine the effects of lipopolysaccharide (LPS)-induced prostatic inflammation on these processes. MATERIALS AND METHODS: Sprague-Dawley rats were divided into three groups: control, 24 hours LPS-inflammation, and 24 hours LPS + antibody against tumor necrosis factor (anti-TNF). 35S-methionine was perifused in vivo around ventral prostate ducts for 3 hours. Ductal fluid (DF) was collected by micropuncture and ductal extract (DE) was collected by tissue homogenization. DE and DF were then subjected to SDS-PAGE and autoradiography. Densitometric analysis of gels and autoradiograms was used to compare protein synthesis (total DE 35S-proteins) and protein secretion (DF 35S-proteins) among the three groups. RESULTS AND CONCLUSIONS: The method proved to be effective for studying prostatic protein synthesis and secretion in vivo. LPS-induced inflammation caused an increase in total 35S-proteins in both the DE and the DF when compared with controls. There were significant increases in both the total number of proteins produced as well as the densitometric quantity of protein in the inflamed group. Some specific prostatic proteins were also upregulated by inflammation. The addition of anti-TNF did not significantly alter inflammation-induced protein synthesis or secretion at the time/dose studied. PMID- 10379796 TI - Bladder infection in the menopausal monkey. AB - PURPOSE: The highest incidence of urinary tract infection in females occurs in elderly women. This study was done to determine whether this is due to the declining immune response that occurs during advancing age, or the menopausal state in the aged. MATERIALS AND METHODS: Adult female monkeys (average age 19 years) were studied, half being subjected to bilateral oophorectomy to produce the menopause. In addition, old females (average age 29 years) already at menopause were studied before and after hormonal replacement with estradiol and progesterone. Bacterial adherence to vaginal cells was studied prior to and after urethral infection with E. coli. Plasma estradiol and progesterone levels were done, as well as white blood counts, plasma cytokine assays and serum antibody titers. RESULTS: Bacteriuria was not prolonged, nor was there a significant difference in bacterial adherence to vaginal cells due to menopause. Interleukin 1 levels were depressed after surgical menopause but not as much as found in the old menopausal females and this low level was not corrected by hormonal replacement. The initial interleukin-2 levels were higher after spontaneous menopause, but the increasing plasma levels seen in cycling animals after infection did not occur in the aged menopausal females following infection even after hormone replacement. The antibody titers to the E. coli infection showed a trend to a lessened response to infection after menopause but were not significantly decreased. CONCLUSIONS: The deficient Il-1, Il-2 and antibody response following infection was not corrected by hormone replacement and thus appears to be due to aging rather than lack of female hormones. These facts may be explained by the T cell senescence known to occur in aged individuals. PMID- 10379797 TI - Are nuclear texture features a suitable tool for predicting non-organ-confined prostate cancer? AB - PURPOSE: We investigated the possibility of determining organ confinement of prostate cancer using multiple nuclear texture features determined by fully automated high resolution image analysis combined with preoperative serum PSA levels. MATERIALS AND METHODS: The study population consisted of 145 patients (61 organ confined and 84 non-organ-confined cases). Nuclear texture features were determined using single cell preparations of radical prostatectomy specimens. Nuclear texture features were extracted and analyzed by multivariate logistic regression analysis in order to build a classifier for distinguishing between organ confined and non-organ-confined tumors. The classifier was designed in a cell by cell model and tested on a case by case analysis. RESULTS: The predictive probability of the trained classifier in the cell by cell analysis had a sensitivity of 63%, a specificity of 53%, a positive predictive value of 75% and a negative predictive value of 38% and an area under the ROC curve of 0.58. In the case by case analysis the sensitivity was 70%, the specificity was 54%, positive predictive value 78%, negative predictive value 74%, area under the ROC curve 0.62. When preoperative PSA was included in the algorithm, sensitivity raised to 80%, specificity to 60%, the positive predictive value raised to 79%, the negative predictive value to 52% and the area under the ROC curve to 0.70. CONCLUSIONS: In contrast to former studies using tissue sections, our results suggest that nuclear texture features extracted from single cell preparations cannot be used as a reliable parameter for the determination of organ confinement in prostatic adenocarcinomas. PMID- 10379798 TI - The CAG repeat within the androgen receptor gene and benign prostatic hyperplasia. PMID- 10379800 TI - Activation for CNS circuits producing a neurogenic cystitis: evidence for centrally induced peripheral inflammation. PMID- 10379799 TI - Rectovestibular fistula with absent vagina: a unique anorectal malformation. PMID- 10379801 TI - Identification of enterococci by ribotyping with horseradish-peroxidase-labelled 16S rDNA probes. AB - Enterococci are frequently associated with hospital-acquired infection. Identification of enterococci using conventional biochemical tests are often tedious to perform in a routine diagnostic laboratory and may give equivocal results. This study evaluates the usefulness of ribotyping by DNA hybridisation to identify 68 members of the bacterial genus Enterococcus characterised by a conventional test scheme. DNA probes (830 bp in size) were derived from the 16S rRNA gene of E. coli or E. faecalis by PCR, labelled with horseradish peroxidase and used in Southern blot hybridisations of enterococcal DNA digested with EcoRI. Unique ribotypes were obtained for 11 different species using 12 Enterococcus type strains. Ribotyping identified 44 E. faecalis isolates, 19 E. faecium isolates, two E. durans isolates and one E. avium isolate in concordance with results of the biochemistry tests. Two isolates that had ribotype patterns identical to the E. faecium type strain were unable to be definitively identified by biochemical tests. The results show that ribotyping is able to differentiate between E. faecium and E. faecalis and may be useful for identifying other enterococci in the hospital setting. In addition, ribotyping using DNA probes and enhanced chemiluminescence is a safe and more reproducible alternative to radiolabelling RNA in a clinical microbiology laboratory. PMID- 10379802 TI - Filtration capture immunoassay for bacteria: optimization and potential for urinalysis. AB - A novel assay utilizing immuno-labeling, filtration, and electrochemistry for the rapid detection of bacteria has been optimized for the detection of Escherichia coli O157:H7. Bacteria were specifically labeled with alkaline phosphatase conjugated polyclonal antibodies and captured on a polycarbonate track-etched membrane filter (0.2 microm pore size). The filter was then placed directly against a glassy carbon electrode, incubated with enzyme substrate, and the product detected by square wave voltammetry. The high speed and capture efficiency of membrane filtration and inherent sensitivity of electrochemical detection produced a 25-min assay with a detection limit of 5 x 10(3) E. coli O157:H7 per ml using a filtration volume of 100 microl (i.e. 500 cells filtered). The labeling, filtration, and electrochemical steps were optimized, and the assay performance using electrochemical and colorimetric detection methods was compared. The assay was used to detect E. coli O157:H7 that was spiked into filter-sterilized urine at clinically relevant concentrations. PMID- 10379803 TI - Application of denaturant gradient gel electrophoresis for the analysis of the porcine gastrointestinal microbiota. AB - The porcine gastrointestinal tract (GIT) microbiota has been studied to increase production efficiency, improve product quality, and help attempt to reduce disease. During the developmental period from birth through weaning, the intestinal microbiota undergoes a rapid ecological succession. There is interest in developing a monitoring technique that allows for analysis of bacterial population levels and shifts within the pig intestine. The objective of this study was to determine if denaturant gradient gel electrophoresis (DGGE) could be effectively applied to measure changes in bacterial populations of the pig GIT, as influenced by age, diet or compartment. Bacterial genetic diversity was determined using DGGE analysis of the V3 region of 16S rDNA PCR products (approximately 200 bp) obtained from primers specific for the domain Bacteria. Protocol development included optimization of: DNA extraction procedures, PCR amplification, removal of PCR artifacts, and optimization of gel preparation and image capture. DGGE analysis revealed diverse bacterial populations between pigs of different ages and among individual gut compartments. Comparison of fecal DNA from different aged pigs revealed several unique PCR product bands indicating the presence of unique bacterial populations. Comparison of different gut compartments demonstrated that bacterial populations were most similar (C, value > 50%) within a single compartment and between adjacent ones. Thus, DGGE can be used to examine bacterial diversity and population shifts in the pig GIT. PMID- 10379804 TI - Comparison of media for the detection of bifidobacteria, lactobacilli and total anaerobes from faecal samples. AB - The interest in functional foods, probiotics and prebiotics requires a proper method to determine specific bacterial groups in the intestinal flora, especially bifidobacteria and lactobacilli. Three media for lactobacilli (MRS, Rogosa, LAMVAB), three media for bifidobacteria (RB, NPNL, Beerens medium) and nine media for total anaerobes have been tested for selectivity and recovery. For total anaerobes Faecal Reinforced Clostridial Agar (FRCA) showed the highest cfu/g, followed by Columbia Blood Agar and BHI Blood agar. There were no significant differences between the media tested. Reduced physiological salt solution was found to be the best dilution medium. For bifidobacteria and lactobacilli samples of human faeces, cat faeces and pig ileal contents were used. Bifidobacteria could reliably be determined on all three media tested in human faeces, but not on pig ileal contents or cat faeces. Absolute counts were highest in human samples. No lactobacilli could be isolated on MRS in either sample, none of the colonies in the countable plates were lactobacilli. For Rogosa over 90% of the colony types observed in human samples were not lactobacilli. For cat faeces this was 58%, but no false positives were observed in the pig ileal samples. For LAMVAB the percentages of false positive colony types were 9, 14 and 0% for human, cat and pig samples. It can be concluded that for bifidobacteria RB and Beerens medium show comparable results, and can be used to quantify bifidobacteria in human faeces, but none of the media tested is suitable for reliably counting bifidobacteria from pig and cat samples. For lactobacilli LAMVAB shows the highest sensitivity. PMID- 10379805 TI - Comparison of API 20NE and Biolog GN identification systems assessed by techniques of multivariate analyses. AB - The increasing use of commercial multitest systems for identification of environmental bacteria creates the problem of how to compare the identification results obtained from different systems. The limited use of species designations in such comparisons is caused by low usage of environmental bacteria in the development of commercial identification schemes. Two multivariate statistical methods, the Mantel's test and the co-inertia analysis, were applied to analyze data derived from the Biolog GN and the API 20NE systems of identification for 50 environmental bacterial strains. We found these two methods to be useful for revealing the relationship between the two sets of numerical taxonomic traits. Both of these methods showed that the distances according to the Biolog GN results between the studied strains were related to those derived from the API 20NE results, despite the differences in the test sets of the two systems. In addition, the co-inertia analysis allowed us to visualise the relationships between classifications of strains derived from the two identification systems and, simultaneously, to estimate the contribution of particular tests to the differentiation of bacterial strains. PMID- 10379806 TI - Comparison of two kinds of Biolog microplates (GN and ECO) in their ability to distinguish among aquatic microbial communities. AB - We compared the abilities of Biolog's GN and ECO plates to distinguish among aerobic and heterotrophic bacterial communities in samples from six aquatic environments. The Biolog system is based on interpreting patterns of sole-carbon substrate utilization indicated by color development in a 96-well microtiter plate. Whether of fresh or saltwater origin, bacterial communities utilized > 95% of substrates in both types of plates. Samples from any one environment exhibited similar time courses of average well color development (AWCD) in both GN and ECO plates. Principal component analysis was performed on data sets resulting from combinations of algorithms (AWCD and curve-integration methods) and levels of color development (end-point and set-point approaches). In all cases, the two types of plates demonstrated an equal capacity to discriminate among the heterotrophic expressions of the six microbial communities. Substantial deviation from an anticipated 1:1 correspondence occurred when color development of 25 substrates common to both types of plates was compared. The discrepancies likely are related to the different formulations of low-nutrient media in GN and ECO plates. PMID- 10379807 TI - Detection of total and hemolysin-producing Vibrio parahaemolyticus in shellfish using multiplex PCR amplification of tl, tdh and trh. AB - Vibrio parahaemolyticus is an important human pathogen which can cause gastroenteritis when consumed in raw or partially-cooked seafood. A multiplex PCR amplification-based detection of total and virulent strains of V. parahaemolyticus was developed by targeting thermolabile hemolysin encoded by tl, thermostable direct hemolysin encoded by tdh, and thermostable direct hemolysin related trh genes. Following optimization using oligonucleotide primers targeting tl, tdh and trh genes, the multiplex PCR was applied to V. parahaemolyticus from 27 clinical, 43 seafood, 15 environmental, 7 strains obtained from various laboratories and 19 from oyster plants. All 111 V. parahaemolyticus isolates showed PCR amplification of the tl gene; however, only 60 isolates showed amplification of tdh, and 43 isolates showed amplification of the trh gene. Also, 18 strains showed amplification of the tdh gene, but these strains did not show amplification of the trh gene. However, one strain exhibited amplification for the trh but not the tdh gene, suggesting both genes need to be targeted in a PCR amplification reaction to detect all hemolysin-producing strains of this pathogen. The multiplex PCR approach was successfully used to detect various strains of V parahaemolyticus in seeded oyster tissue homogenate. Sensitivity of detection for all three target gene segments was at least between 10(1)-10(2) cfu per 10 g of alkaline peptone water enriched seeded oyster tissue homogenate. This high level of sensitivity of detection of this pathogen within 8 h of pre enrichment is well within the action level (10(4) cfu per 1 g of shell stock) suggested by the National Seafood Sanitation Program guideline. Compared to conventional microbiological culture methods, this multiplex PCR approach is rapid and reliable for accomplishing a comprehensive detection of V. parahaemolyticus in shellfish. PMID- 10379808 TI - Green fluorescent protein-based direct viable count to verify a viable but non culturable state of Salmonella typhi in environmental samples. AB - The gfp-tagging method and lux-tagging method were compared to select a better method for verifying a viable but nonculturable (VBNC) state of bacteria in the environment. An environmental isolate of Salmonella typhi was chromosomally marked with a gfp gene encoding green fluorescent protein (GFP). The hybrid transposon mini-Tn5 gfp was transconjugated from E. coli to S. typhi. Using the same method, S. typhi was chromosomally marked with luxAB genes encoding luciferase. The survival of gfp-tagged S. typhi introduced into groundwater microcosms was examined by GFP-based plate count, total cell count, and a direct viable count method. In microcosms containing lux-tagged S. typhi, luminescence based plate count and the measurement of bioluminescence of each microcosm sample were performed. In microcosms containing lux-tagged S. typhi, viable but nonculturable cells could not be detected by using luminometry. As no distinguishable luminescence signals from the background signals were found in samples containing no culturable cells, a VBNC state of S. typhi could not be verified in lux-based systems. However, comparison between GFP-based direct viable counts and plate counts was a good method for verifying the VBNC state of S. typhi. Because GFP-based direct viable count method provided a direct and precise estimation of viable cells of introduced bacteria into natural environments, it can be used for verifying the VBNC state of bacteria in environmental samples. PMID- 10379809 TI - Cytogenetic effects of ethylene oxide, with an emphasis on population monitoring. AB - Cytogenetic assays are an integral component of the battery of short-term assays that are used for the hazard identification component of a cancer risk assessment. The protocol for the conduct of such assays for maximal sensitivity for detecting clastogenicity has to be attendant to the mechanism of induction of the endpoint being assessed and the fact that several aberration types are cell lethal necessitates that analysis be for cells at their first posttreatment metaphase. Cytogenetic assays for human populating monitoring have been used for predicting potential for carcinogenicity in humans. However, the assays as typically conducted are not appropriate for chronic exposures because nontransmissible alterations are assessed. The use of fluorescent in situ hybridization (FISH) techniques for the assessment of transmissible changes such as reciprocal translocations are required to make population monitoring studies interpretable, and for removing some of the concern over the influence of confounders on outcome. The database for the cytogenetic effects of ethylene oxide in vitro and in vivo, with an emphasis on human population monitoring, has been critically reviewed. Based on the endpoints studied, the size of the study groups, the information on exposure, the nature of any exposure response data, and the possible influence of confounders (i.e., control matching), it is concluded that acute, high exposures to ethylene oxide with sampling shortly (a few days) after exposure can be detected by increases in chromosome aberrations or SCE in peripheral lymphocytes. Such increases are indicators of exposure to a genotoxic chemical and not predictors of subsequent adverse health effects to individuals. The effect of chronic and/or low level (less than about 25 ppm) exposures cannot be reliably evaluated using current methods. The use of FISH, for example, for assessing reciprocal translocation frequencies (as a measure of transmissible events) will greatly improve the ability to detect chronic exposures to clastogenic chemicals. PMID- 10379810 TI - The toxicology of hydroquinone--relevance to occupational and environmental exposure. AB - Hydroquinone (HQ) is a high-volume commodity chemical used as a reducing agent, antioxidant, polymerization inhibitor, and chemical intermediate. It is also used in over-the-counter (OTC) drugs as an ingredient in skin lighteners and is a natural ingredient in many plant-derived products, including vegetables, fruits, grains, coffee, tea, beer, and wine. While there are few reports of adverse health effects associated with the production and use of HQ, a great deal of research has been conducted with HQ because it is a metabolite of benzene. Physicochemical differences between HQ and benzene play a significant role in altering the pharmacokinetics of directly administered when compared with benzene derived HQ. HQ is only weakly positive in in vivo chromosomal assays when expected human exposure routes are used. Chromosomal effects are increased significantly when parenteral or in vitro assays are used. In cancer bioassays, HQ has reproducibly produced renal adenomas in male F344 rats. The mechanism of tumorigenesis is unclear but probably involves a species-, strain-, and sex specific interaction between renal tubule toxicity and an interaction with the chronic progressive nephropathy that is characteristic of aged male rats. Mouse liver tumors (adenomas) and mononuclear cell leukemia (female F344 rat) have also been reported following HQ exposure, but their significance is uncertain. Various tumor initiation/promotion assays with HQ have shown generally negative results. Epidemiological studies with HQ have demonstrated lower death rates and reduced cancer rates in production workers when compared with both general and employed referent populations. Parenteral administration of HQ is associated with changes in several hematopoietic and immunologic endpoints. This toxicity is more severe when combined with parenteral administration of phenol. It is likely that oxidation of HQ within the bone marrow compartment to the semiquinone or p benzoquinone (BQ), followed by covalent macromolecular binding, is critical to these effects. Bone marrow and hematologic effects are generally not characteristic of HQ exposures in animal studies employing routes of exposure other than parenteral. Myelotoxicity is also not associated with human exposure to HQ. These differences are likely due to significant route-dependent toxicokinetic factors. Fetotoxicity (growth retardation) accompanies repeated administration of HQ at maternally toxic dose levels in animal studies. HQ exposure has not been associated with other reproductive and developmental effects using current USEPA test guidelines. The skin pigment lightening properties of HQ appear to be due to inhibition of melanocyte tyrosinase. Adverse effects associated with OTC use of HQ in FDA-regulated products have been limited to a small number of cases of exogenous ochronosis, although higher incidences of this syndrome have been reported with inappropriate use of unregulated OTC products containing higher HQ concentrations. The most serious human health effect related to HQ is pigmentation of the eye and, in a small number of cases, permanent corneal damage. This effect has been observed in HQ production workers, but the relative contributions of HQ and BQ to this process have not been delineated. Corneal pigmentation and damage has not been reported at current exposure levels of <2 mg/m3. Current work with HQ is being focused on tissue specific HQ-glutathione metabolites. These metabolites appear to play a critical role in the renal effects observed in F344 rats following HQ exposure and may also be responsible for bone marrow toxicity seen after parenteral exposure to HQ or benzene-derived HQ. PMID- 10379811 TI - Cellular and antibody responses to the Plasmodium falciparum heat shock protein Pf72/HSP70 during and after acute malaria in individuals from an endemic area of Brazil. AB - Proliferative and antibody responses to three synthetic peptides corresponding to Pf72/ HSP70 were followed-up in acute malaria patients from an endemic area of Brazil. In vitro lymphocyte responsiveness to all peptides was relatively low and short-lived and there was a considerable variation in the frequency and magnitude of the individual lymphoproliferative response to the peptides at different periods after the onset of infection. Although 96% of the patients had IgG antibodies to crude Plasmodium falciparum asexual blood stage antigens, specific IgG antibody responses to the peptides varied from 12.5 to 40% according to the tested peptides. No significant difference was observed in the proliferative or antibody responses to the peptides between individuals that remained parasitemic after treatment and those that recovered from malaria infection. The different frequencies of proliferative responses in peripheral blood T cells on different occasions after the onset of their infection show that, in order to be informative, evaluation of the in vitro cellular immune response to peptides requires longitudinal studies in which each individual is tested repeatedly. PMID- 10379812 TI - Improved latex agglutination test for detection of antibodies in serum and cerebrospinal fluid of Trypanosoma brucei gambiense infected patients. AB - A rapid latex agglutination test (LATEX/T. b. gambiense) for detection of antibodies in patients infected with Trypanosoma brucei gambiense is presented. The reagent is coated with a mixture of three variable surface antigens of bloodstream form trypanosomes. Two hundred and forty sera and 79 CSF samples from patients with parasitologically confirmed trypanosome infection along with 173 sera and 38 CSF samples from non-trypanosomiasis patients have been tested. At 1:16 serum dilution, test specificity was 99%, while sensitivity ranged from 83.8 to 100% depending on the geographical origin of the samples. Undiluted CSF samples from non-trypanosomiasis and from first stage patients scored negative while 42 out of 66 CSF samples from second stage patients were positive. Stability and reproducibility of the lyophilized reagent were excellent. PMID- 10379813 TI - PCR-ELISA for diagnosis of mucocutaneous leishmaniasis. AB - In this work we demonstrate that the PCR-ELISA technique is sufficiently sensitive and specific for use as a diagnostic test in cases of mucocutaneous leishmaniasis. DNA was extracted from cultures of Leishmania braziliensis, Leishmania infantum, Leishmania tropica, Leishmania mexicana, Trypanosoma cruzi, and blood samples from individuals who presented a clinical diagnosis of leishmaniasis as well as from healthy individuals. The DNA was PCR amplified and the product obtained was hybridised with a biotin-labelled probe, the sequence of which was designed in our laboratory. The result of the hybridisation was visualised by means of an ELISA technique using antifluorescein antibody labelled with alkaline phosphatase and p-nitrophenylphosphate (pNFF) as chromogen. The optical density of the products of the pNFF hydrolysis was quantified in a spectrophotometer at a wavelength of 405 nm. Using this technique the percentage of detection was 83.3% in blood samples from patients clinically diagnosed as having mucocutaneous leishmaniasis. No false positive results were obtained. PMID- 10379814 TI - Human and porcine Taenia solium infection in a village in the highlands of Cusco, Peru. The Cysticercosis Working Group in Peru. AB - A serological survey was performed using the enzyme-linked immunoelectrotransfer blot assay (EITB) in a village in the highlands of Peru where there are three distinct but close neighborhoods, to determine if there is a direct relationship between human and porcine Taenia solium infection. One hundred and eight out of 365 individuals were sampled, and 14 were seropositive (human seroprevalence 13%). Most seropositive individuals were neurologically asymptomatic. Thirty eight out of 89 sampled pigs (43%) were seropositive. There was a clear geographical clustering of cases, and positive correlation between human and porcine seroprevalence found when comparing the three neighborhoods. Cysticercosis is an important cause of neurological morbidity in most developing countries, and control/eradication trials are now being increasingly applied. Porcine serology provides an appropriate indicator of T. solium environmental contamination and should be used to estimate the risk of infection when evaluating control measures. PMID- 10379815 TI - Leishmania donovani: cellular and humoral immune responses in Indian langur monkeys, Presbytis entellus. AB - We have previously reported that disease mimicking human visceral leishmaniasis can be established in Presbytis entellus, the Indian langur monkey, following a single intravenous challenge of 10(8) Leishmania donovani amastigotes. In the present report, infection was assessed in monkeys infected intravenously with a single dose of 10(8) amastigotes (HDA group), three weekly doses of 10(7) amastigotes (LDA group) and three weekly doses of 5 x 10(7) promastigotes (HDP group). Typical clinical infection was established in all three groups with significant parasite load. There was a gradual and sustained rise in anti leishmania specific immunoglobulin G response, and a severe fall in the lymphoproliferative response to the T cell mitogens PHA and Con A by day 80 post infection (p.i.). The antibody level remained elevated until death in monkeys of the HDA and HDP groups; the T-cell responses showed a recovery prior to death. T cell responses to leishmania antigen, however, could not be demonstrated in any of these monkeys prior to death. One monkey of the LDA group survived for 155 days and two monkeys spontaneously eradicated the infection. Surprisingly, one monkey of the HDA group also achieved spontaneous cure. In the three monkeys which eradicated infection spontaneously, the antibody level declined to baseline levels on day 180 p.i. with a well demonstrable antigen specific lymphoproliferative response; no parasites could be demonstrated in splenic aspirates by direct examination of culture. These data demonstrate that disease severity may be the function of the total inoculum dose rather than the stage of the parasite and that the immunological responses in the Indian langur model parallel the reported changes in human visceral leishmaniasis. This makes the langur a potentially useful model for the evaluation of candidate anti leishmanial drugs and vaccines. PMID- 10379816 TI - Extension of the prophylactic effect of isometamidium against trypanosome infections in cattle using a biodegradable copolymer. AB - Two trials were carried out in order to compare the prophylactic effect of a subcutaneously implanted sustained release device (SRD) containing a mixture of a biodegradable copolymer, poly(caprolactone-co-L-lactide), and isometamidium (ISMM) with that obtained after intramuscular injection of the drug. In a first experiment under controlled conditions, two groups of cattle were treated with 0.5 mg/kg isometamidium either as a SRD or intramuscularly (i.m.), and exposed at monthly intervals to Glossina morsitans morsitans infected with Trypanosoma congolense. The average protection period was at least 24 months in the SRD treated against 5.7 months in the i.m. treated group. Using an ISMM enzyme-linked immunosorbent assay, the drug could be detected until 140 days post-treatment in the latter group, whereas in the former group, traces of the drug were detectable until 330 days after treatment. Furthermore, a field trial was carried out at the Madina Diassa ranch in Mali involving three groups of N'Dama cattle, each containing 23 or 24 animals. Two groups were treated with 1 mg/kg ISMM either as a SRD or i.m. and a third group served as untreated control. Twelve months after treatment, the cumulative infection rates were 56.5, 87.8 and 91.6% in the SRD implanted, the i.m. treated and the control groups, respectively. The ISMM concentrations were slightly lower than in the laboratory trial, but the overall pattern of drug disappearance from the sera of the SRD treated cattle was very similar in both trials. Statistical analysis showed that the incidence of trypanosomiasis was significantly lower in the SRD treated than in the i.m. treated group. PMID- 10379817 TI - Characterization of porcine and ovine Oesophagostomum spp. by isoenzymatic patterns and restriction-fragment-length polymorphisms (RFLPs). AB - Four different morphological and biometrical populations of Oesophagostomum have been identified, using classical taxonomy methods, from Sus scrofa domestica (pigs): O. dentatum (Od), O. quadrispinulatum (Oq), O. granatensis (Og), and a fourth population, including individuals with morphological and biometric parameters overlapping these three species that were clasified as Oesophagostomum sp. The G6PD and MDH isoenzymatic patterns did not discriminate between the three species, while GPI showed a diagnostic isoenzymatic pattern for Oq. Og showed identical G6PD, GPI and MDH isoenzymatic pattern as Od. Furthermore, after rDNA amplification by polymerase chain reaction (PCR), the uncut PCR product showed that the ITS2 of these three species had a similar size of 320 base pairs (bp). Restriction-fragment-length polymorphisms (RFLP) were analyzed after digestion of the ITS2 with 13 different restriction enzymes. After electrophoretic separation of the digested PCR products, only one unique differentiating pattern of bands was observed for Od and Oq. This was when Sau3AI was used, while Og showed an identical band pattern to Od. Thus, our studies provided no evidence for the existence of Og and Od as differentiated populations. O. venulosum was isolated from sheep and goat; G6PD and MDH isoenzymatic patterns discriminated this species from porcine species of Oesophagostomum. The ITS2 region appeared as a different band of 380 bp from those observed for porcine Oesophagostomum species. PMID- 10379818 TI - Blood coagulation factor XIII: structure and function. PMID- 10379819 TI - L-arginine modulates aggregation and intracellular cyclic 3,5-guanosine monophosphate levels in human platelets: direct effect and interplay with antioxidative thiol agent. AB - Platelet nitric oxide is involved in the control of aggregability via cyclic 3',5'-guanosine monophosphate synthesis. Since L-arginine provides a guanidino nitrogen group for nitric oxide synthesis through nitric oxide synthase activity, we tried to clarify whether an increased availability of this amino acid can directly modulate the response of human platelets. In our conditions, L-arginine (at 100-6000 micromol/L) was able to influence the response of human platelets stimulated with adenosine 5-diphosphate and collagen both in PRP and in whole blood. The anti-aggregating effect was not present when D-arginine was used. Permeabilized platelets exhibited an increased sensitivity to L-arginine. Also, an increased availability of Ca2+ enhanced L-arginine effect. L-arginine (at 120 500 micromol/L) increased cyclic 3',5'-guanosine monophosphate levels in resting platelets; the amino acid also determined an increase of cyclic 3',5'-guanosine monophosphate in platelets at the end of adenosine 5-diphosphate-induced aggregation. Nitric oxide synthase inhibitor N(G)-monomethyl-L-arginine prevented L-arginine effects on aggregation and cyclic 3',5'-guanosine monophosphate synthesis. Phosphodiesterase III inhibitor milrinone and antioxidative thiol N acetyl-L-cysteine enhanced the effect of L-arginine on cyclic 3',5'-guanosine monophosphate. In conclusion, L-arginine exerts inhibitory effects on human platelet response through a nitric oxide-dependent synthesis of cyclic 3',5' guanosine monophosphate. A positive interplay on platelet response between L arginine and milrinone or antioxidative thiol N-acetyl-L-cysteine was evidenced. PMID- 10379820 TI - Effects of lysophosphatidic acid on proliferation and cytosolic Ca++ of human adult vascular smooth muscle cells in culture. AB - Lysophosphatidic acid (LPA) is a lipid mediator generated by activated platelets and having various effects on numerous cell types. We investigated some effects of 1-oleyl LPA on vascular smooth muscle cells cultured from adult human normal arteries. At micromolar concentrations, LPA induced a mitogenic effect ([3H] thymidine incorporation and cell proliferation) on quiescent cells, without an additional growth factor being required. This effect was equipotent to that of 10% fetal calf serum, and it was accompanied by early (5 minutes) and late (1-3 hours) phosphorylation of mitogenactivated protein kinase. LPA inhibited cell migration through collagen coated membranes, with or without platelet-derived growth factor BB as chemoattractant. LPA induced a typical biphasic Ca2+ signal response made up of a rapid first phase due to Ca2+ release from intracellular stores followed by a second wave due to external Ca2+ influx. These findings support the proposal that LPA released from activated platelets is a mediator for smooth muscle cell response at the site of vessel injury in humans. PMID- 10379821 TI - Chronic oral defibrotide counteracts hypercholesterolemia noxious effects on cardiovascular function in the rabbit. AB - The aim of the present work was to assess if the cardioprotective drug defibrotide could counteract the hypercholesterolemia noxious effects on cardiovascular function. Aortas and hearts from normal- or cholesterol-fed rabbits, treated or not with chronic oral defibrotide (100 mg/kg/day) for 45 days, were used in in vitro tests throughout the experiment. Hypercholesterolemia worsened: aorta stickiness toward polymorphonuclear leukocytes, aorta relaxation to acetylcholine, heart left ventricular end-diastolic pressure and coronary perfusion pressure, heart left ventricular diastolic pressure, acetylcholine and endothelin-1 activity on coronary perfusion pressure, and heart generation of 6 Keto-prostaglandin F1alpha. Oral defibrotide counteracted and/or obliterated the above hypercholesterolemia noxious effects. Particularly, oral defibrotide counteracted the parameters associated with early endothelial cell disfunction: that is, increased adherence of leukocytes to endothelium and endothelial vasorelaxation induced by acetylcholine, which acts through the release of endothelium-derived relaxing factor. These activities of defibrotide are probably exerted through the increased generation of prostacyclin. The fact that acetylcholine induced vasorelaxation is partially protected by oral defibrotide points to a partial rescue of endothelial ability to generate endothelium-derived relaxing factor, as acethylcoline acts through the release of endothelium-derived relaxing factor, by defibrotide itself. Defibrotide's endothelial protection could, in turn, explains why defibrotide protected cardiovascular function. This is not surprising as, in a few cases, endothelial dysfunction, observed in hypercholesterolemia, was found to be prevented or reversed, pharmacologically, by PN-2001-10, a calcium channel blocker, dipyridamole, and lovastatin. PMID- 10379822 TI - Postnatal distribution of cpp32/caspase 3 mRNA in the mouse central nervous system: an in situ hybridization study. AB - Apoptotic cell death is a major feature of the developing nervous system and of certain neurodegenerative diseases. Various gene effectors and repressors of this type of cell death have been identified. Among them, bcl-xl and bax, which encode for antiapoptotic and proapoptotic proteins, respectively, play major roles during development. The gene cpp32 encodes for the caspase 3 cysteine protease and is a critical mediator of cell death during embryonic development in the mammalian brain. To gain insight into the possible implications of these cell death genes during the postnatal development, we investigated the expression of bax, bcl-xl, and cpp32 mRNAs by in situ hybridization in the mouse brain from birth to adulthood. Whereas bax and bcl-xl mRNAs were expressed widely in neonates and adult mice, our results showed that cpp32 mRNA levels were decreased strongly from 12 postnatal days. From 1 postnatal day to 12 postnatal days, cpp32 mRNA was expressed ubiquitously in all brain nuclei, including areas where neurogenesis occurred. A positive correlation between areas displaying high levels of mRNA and apoptotic nuclei also was shown. In the adult, cpp32 mRNA was restricted to the piriform and entorhinal cortices, the neocortex, and to areas where neurogenesis is observed (e.g., olfactory bulb and dentate gyrus). The same pattern of expression was observed in adult mice over-expressing the antiapoptotic protein Bcl-2. These results demonstrate that the expression of cpp32 mRNA is highly regulated during the mouse postnatal period, leading to a specific distribution in the adult central nervous system. Moreover, the prevention of cell death by Bcl-2 likely is not linked to the regulation of caspase mRNA levels. PMID- 10379823 TI - Spinal organization and steroid sensitivity of motoneurons innervating the pubococcygeus muscle in the male rat. AB - Male rat motoneurons innervating the pubococcygeus muscle were located in the ventral nucleus of lamina IX at the sixth lumbar (L6) and first sacral (S1) spinal cord segments. Retrograde labeling with horseradish peroxidase-wheat germ agglutinin was transported up to second-order dendrites and revealed that these motoneurons have a "U-shaped arborization" of dendrites toward the intermediolateral and intermediomedial nuclei area of lamina VII. This dendritic organization makes a wide "final common path" that probably integrates afferent information from several sources, accounting for the participation of the pubococcygeus muscle in autonomic and somatic processes, such as those related to micturition and reproduction. Castration produced a decrement in the morphometry of these motoneurons. A main effect was a decrement in dendritic length. Steroid replacement indicated that testosterone and estradiol, but not dihydrotestosterone, are able to induce a recovery of morphometric alterations. However, estrogen induced recovery after 2 weeks of treatment, whereas testosterone took 4 weeks. Thus, it is proposed that supraspinal aromatization of testosterone in the male central nervous system might be an important process for the appropriate organization of the pubococcygeus muscle motoneurons and that estradiol seems to need a shorter time of action than testosterone because of differential up-regulation and down-regulation of steroid receptors. PMID- 10379824 TI - Organization of projections from the anterior pole of the nucleus reticularis thalami (NRT) to subdivisions of the motor thalamus: light and electron microscopic studies in the rhesus monkey. AB - Projections to the motor-related thalamic nuclei from the anterior pole of the reticular thalamic nucleus (NRT) were studied after injections of biotinylated dextran amine and wheat germ agglutinin conjugated horseradish peroxidase at light and electron microscopic levels, respectively. Each injection resulted in anterograde labeling in the three subdivisions of the ventral anterior nucleus (pars parvicellularis, VApc; pars densicellularis, VAdc; and pars magnocellularis, VAmc) and in the ventral lateral nucleus (VL). NRT fibers had beaded shapes and coursed in a posterior direction giving rise to relatively diffuse terminal plexuses. The average size of the beads (0.7 microm2) and their density per 100 microm of fiber length (23.7-25.7) were similar between the nuclei studied. At the electron microscopic level, anterogradely labeled boutons displayed positive immunoreactivity for gamma-aminobutyric acid (GABA), contained pleomorphic synaptic vesicles, and formed relatively long (approximately 0.4 microm) symmetric synaptic contacts. Usually, a single terminal formed synapses on more than one postsynaptic structure. Synaptic contacts were on projection and local circuit neurons and targeted mainly their distal dendrites. In the VAmc, synapses on local circuit neurons composed 48% of the total sample, in the VAdc/VApc and in the VL the proportion was higher, 65% and 62%, respectively. The results suggest that the input from the anterior pole of the monkey reticular nucleus to the motor-related thalamic nuclei is organized differently from what is known on the organization of connections of NRT with sensory thalamic nuclei in other species in that the terminal fields of individual fibers are diffuse rather than focal and that at least 50% of synapses are established on GABAergic local circuit neurons. PMID- 10379825 TI - Dendritic bias of neurons in rat somatosensory cortex associated with a functional boundary. AB - Sensory information is encoded throughout the central nervous system by activation of specific groups of neurons. Neurons encoding information from a particular modality are grouped together and constitute an ordered neural representation or "map" of the stimulus. The organization of these representations is not static, but is capable of significant alteration in response to changes in the patterns of inputs delivered to the cortex in the appropriate behavioral context. Therefore, understanding the basic mechanisms that account for discontinuities in cortical representations are important for understanding both information processing in the cortex and plasticity of cortical organization. It is clear that both anatomic and physiologic mechanisms underlie both the genesis and the plasticity of these representations; however, their exact contributions are not fully understood. To examine neuronal anatomy around a representational border in rat primary somatosensory cortex (S1), a novel in vivo/in vitro preparation was used in which the location of the border between the forepaw and lower jaw representations in rat S1 was determined electrophysiologically and marked by dye iontophoresis in vivo. By using in vitro slices from the region in which this border was marked, the morphologies of single cortical layer 2/3 neurons close to and far from the border were determined by intracellular injection of biocytin. Neurons close to the border had dendritic arbors that were significantly biased away from the border; neurons far from the border did not. This bias was due to a decrease in the number of neurites that specifically crossed the border with a concomitant increase in other near-border parts of the neuron, consistent with the ideas that patterns of activity are important for neurite outgrowth and that neurons maintain a relatively constant total neurite extent. These findings confirm the close association of cortical anatomy and physiology and illustrate their relationships with cortical representational discontinuities. PMID- 10379826 TI - Tyrosine hydroxylase-immunoreactive elements in the human globus pallidus and subthalamic nucleus. AB - In contrast to the well-established dopaminergic innervation of the neostriatum, the existence of dopaminergic innervation of the subthalamic nucleus and globus pallidus is controversial. In the present study, tyrosine hydroxylase (TH) immunoreactive elements were observed by light microscopy after antigen retrieval in the subthalamic nucleus and in the internal and external segments of the globus pallidus in postmortem human brain. Small islands of apparent neostriatal tissue with abundant arborization of fine, TH-immunoreactive axons in the vicinity of calbindin-positive small neurons resembling neostriatal medium spiny neurons were present in the external segment of the globus pallidus. Large numbers of medium-large, TH-immunoreactive axons were observed passing above and through the subthalamic nucleus and through both pallidal segments; these are presumed to be axons of passage on their way to the neostriatum. In addition, fine, TH-immunoreactive axons with meandering courses, occasional branches, and irregular outlines, morphologically suggestive of terminal axon arborizations with varicosities, were seen in both pallidal segments, including the ventral pallidum, and the subthalamic nucleus, consistent with a catecholaminergic (probably dopaminergic) innervation of these nuclei. This finding suggests that, in Parkinson's disease and in animal models of this disorder, loss of dopaminergic innervation might contribute to abnormal neuronal activation in these three nuclei. PMID- 10379827 TI - Laryngeal afferent stimulation enhances Fos immunoreactivity in periaqueductal gray in the cat. AB - The main functions of the larynx are protection of the airways, respiration, and vocalization. Previous studies have suggested a link between the mechanisms controlling vocalization and afferent feedback from the larynx. We inquired whether stimulation of the laryngeal afferents that run in the internal branch of the superior laryngeal nerve (ISLN) activates neurons of the periaqueductal gray (PAG), a midbrain region implicated in vocalization. We counted the number of neurons expressing Fos, the protein product of the immediate early gene c-fos, in the PAG. The counts were done both in experimental cats after electrical stimulation of the ISLN and nonstimulated controls. We also investigated the possible presence of nitric oxide synthase, an enzyme that synthesizes nitric oxide, in PAG neurons that respond to laryngeal afferent stimulation by double labeling for reduced nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase and Fos. Fos expression was significantly greater (P < or = 0.00714) in the lateral and dorsolateral regions of the PAG in the experimental group than in the controls. The Fos-immunoreactive neurons did not contain NADPH-diaphorase, a marker for nitric oxide synthase. Our study suggests that laryngeal afferent stimulation activates neurons in discrete longitudinal columns of the PAG including the regions that have previously been shown to be involved in vocalization, and that these neurons do not contain nitric oxide synthase. PMID- 10379828 TI - Novel membranous structures in apical and basal compartments of inner hair cells. AB - Postfixation with a ferrocyanide-osmium tetroxide solution preserved a dense network of canaliculi extending from the apical to the upper lateral plasma membrane in cochlear inner hair cells (IHCs). Numerous Golgi bodies intermingled with this apical canalicular reticulum (CR). Osmium-ferrocyanide treatment also disclosed several previously unreported structures below the IHC nucleus. The first consisted of stacks of six or eight and sets of three parallel cisternae of rough endoplasmic reticulum spanning between clustered mitochondria. Some parallel cisternae ended with segmentation where they contacted mitochondria, and others terminated by transforming into blebs or continuing into canaliculi. A second feature was comprised of a complex of segmented cisternae and branching canaliculi with clustered mitochondria. Branching minicanaliculi with associated vesicles neighbored the complexes. A fourth entity consisted of synaptic-like vesicles that largely filled the subnuclear cytosol and congregated at synapses. An additional infranuclear structure was composed of slender canaliculi that collected near or streamed to plasmalemma, often next to a synapse. A paradoxical absence of rough endoplasmic reticulum above and Golgi zones below the nucleus provided evidence of atypical mechanisms for generating the membrane in CR and forming synaptic vesicles. The observations offer the view that IHCs are compartmentalized into an apical mechanoreceptor half and a basal half that affects neurotransmission. The apical CR provided a possible structural basis for sequestering the K+ known to influx apically and for directing its diffusion to the site of known efflux across the lateral plasmalemma. The codistribution of parallel cisternae, canalicular-mitochondrial complexes, and synaptic-like vesicles, all of which are unique to IHCs, implicated the cisternae and complexes in the genesis of the vesicles. PMID- 10379829 TI - Ovarian hormones differentially influence immunoreactivity for dopamine beta- hydroxylase, choline acetyltransferase, and serotonin in the dorsolateral prefrontal cortex of adult rhesus monkeys. AB - Recent studies have shown that ovariectomy reduces, and subsequent hormone replacement restores the density of axons immunoreactive for tyrosine hydroxylase in the dorsolateral prefrontal cortex of adult female rhesus monkeys. The present study indicates that three additional extrathalamic frontal lobe afferents are also sensitive to changes in the ovarian hormone environment. Specifically, the combination of hormone manipulation with qualitative and quantitative analysis of immunocytochemistry for dopamine beta-hydroxylase, choline acetyltransferase, and serotonin in the primate prefrontal cortex revealed quantitative responses in both cholinergic and monoaminergic axons to changing ovarian hormone levels. However, whereas ovariectomy produced a modest net decrease in the density of fibers immunoreactive for choline acetyltransferase, this same treatment markedly increased the density of axons immunoreactive for dopamine beta-hydroxylase and for serotonin. Further, the effects of ovariectomy on these afferent systems were differentially attenuated by estrogen verses estrogen plus progesterone hormone replacement. Estrogen was as effective as estrogen plus progesterone in stimulating normal prefrontal immunoreactivity for choline acetyltransferase and dopamine beta-hydroxylase. The dual replacement of estrogen plus progesterone, however, was a much more potent influence than estrogen alone for serotonin immunoreactivity. Thus, ovarian hormones appear to provide stimulation that differentially affects each of four chemically identified extrathalamic prefrontal afferent systems examined to date, and may have roles in maintaining the normal balance and functional interactions between these neurotransmitter systems. PMID- 10379830 TI - Neurochemical organization of inferior pulvinar complex in squirrel monkeys and macaques revealed by acetylcholinesterase histochemistry, calbindin and Cat-301 immunostaining, and Wisteria floribunda agglutinin binding. AB - To investigate whether the inferior pulvinar complex has a common organization in different primates, the chemoarchitecture of the visual thalamus was re-examined in squirrel monkeys (Saimiri sciureus) and macaques (Macaca mulatta). The inferior pulvinar (PI) complex consisted of multiple subdivisions and encompassed the classic PI, and adjacent ventral parts of the lateral and medial pulvinar (PL and PM, respectively). In keeping with nomenclature suggested previously for macaques, the PI subdivisions were termed the posterior, medial, central, lateral, and lateral-shell (PI(P), PI(M), PI(C), PI(L), and PI(L-S)). In both species, PI(P) was intense for calbindin, light for acetylcholinesterase (AChE), and very light for Wisteria floribunda agglutinin (WFA) histochemistry. The PI(M) was calbindin poor, AChE rich, and moderate for WFA. The PI(C) was calbindin intense, lighter for AChE, and exhibited little WFA binding. PI(L) and PI(L-S) contained populations of large calbindin or WFA cells that were more numerous in PI(L-S). Although staining with the monoclonal antibody Cat-301 differed between macaques and squirrel monkeys, the same subdivisions were displayed. Moderately dense, patchy Cat-301 stain was found in PI(M) of macaques, whereas in squirrel monkeys PI(M) was light. Connections of the rostral dorsolateral (DLr) and middle temporal (MT) areas of visual cortex in squirrel monkeys were compared with PI subdivisions revealed by the newer histochemical methods in the same cases. The major connections of DLr were with PI(C) and of MT were with PI(M). PMID- 10379831 TI - Expression of the novel galanin receptor subtype GALR2 in the adult rat CNS: distinct distribution from GALR1. AB - Recent molecular cloning studies by our laboratory and others have identified the existence of a novel rat galanin receptor subtype, GALR2. In the present study, we examined the regional and cellular distribution of GALR2 mRNA in the rat central nervous system (CNS) by in situ hybridization. For comparative purposes, adjacent sections were probed for GALR1 mRNA expression. Our findings indicate that dorsal root ganglia express by far the highest levels of GALR2 mRNA in the rat CNS. Hybridization signal is mainly concentrated over small and intermediate primary sensory neurons. In spinal cord, the large alpha motoneurons of the ventral horn are moderately labeled and several small, but less intensely labeled, cells are scattered throughout the gray matter. In brain sections, the highest levels of GALR2 mRNA are detected in granule cells of the dentate gyrus, in the mammillary nuclei, and in the cerebellar cortex. Moderate levels of GALR2 mRNA are observed in the olfactory bulb, olfactory tubercle, piriform and retrospinal cortices, hypothalamus (namely the preoptic area, arcuate nucleus, and dorsal hypothalamic area), substantia nigra pars compacta, and sensory trigeminal nucleus. Moderate to weak hybridization signal is also present in several other hypothalamic nuclei, specific layers of the neocortex, periaqueductal gray, and several nuclei within the pons and medulla, including locus coeruleus, lateral parabrachial, motor trigeminal, pontine reticular, hypoglossal, vestibular complex, ambiguus, and facial and lateral reticular nuclei. This novel pattern of GALR2 distribution within the rat CNS differs considerably from that of GALR1, suggesting that specific physiologic effects of galanin may be ascribed to the GALR2 galanin receptor subtype. PMID- 10379832 TI - Neuronal, glial, and epithelial localization of gamma-aminobutyric acid transporter 2, a high-affinity gamma-aminobutyric acid plasma membrane transporter, in the cerebral cortex and neighboring structures. AB - Neuronal and glial high-affinity Na+/Cl(-)-dependent plasma membrane gamma aminobutyric acid (GABA) transporters (GATs) contribute to regulating neuronal function. We investigated in the cerebral cortex and neighboring regions of adult rats the distribution and cellular localization of the GABA transporter GAT-2 by immunocytochemistry with affinity-purified polyclonal antibodies that react monospecifically with a protein of 82 kDa. Conventional and confocal laser scanning light microscopic studies revealed intense GAT-2 immunoreactivity (ir) in the leptomeninges, choroid plexus, and ependyma. Weak GAT-2 immunoreactivity also was observed in the cortical parenchyma, where it was localized to puncta of different sizes scattered throughout the radial extension of the neocortex and to few cell bodies. In sections double-labeled with GAT-2 and glial fibrillary acidic protein (GFAP) antibodies, some GAT-2-positive profiles also were GFAP positive. Ultrastructural studies showed GAT-2 immunoreactivity mostly in patches of varying sizes scattered in the cytoplasm of neuronal and nonneuronal elements: GAT-2-positive neuronal elements included perikarya, dendrites, and axon terminals forming both symmetric and asymmetric synapses; nonneuronal elements expressing GAT-2 were cells forming the pia and arachnoid mater; astrocytic processes, including glia limitans and perivascular end feet; ependymal cells; and epithelial cells of the choroid plexuses. The widespread cellular expression of GAT-2 suggests that it may have several functional roles in the overall regulation of GABA levels in the brain. PMID- 10379833 TI - Immunocytochemistry of GABA in the central complex of the locust Schistocerca gregaria: identification of immunoreactive neurons and colocalization with neuropeptides. AB - The central complex is a highly organized neuropil structure in the insect brain and plays a role in motor control and visual orientation. We describe the distribution of gamma-aminobutyric acid (GABA) immunostaining in the central complex of the locust Schistocerca gregaria in an effort to analyze inhibitory neural circuits within this brain area. Antisera against GABA and the GABA synthesizing enzyme glutamic acid decarboxylase resulted in identical patterns of immunostaining. Cell counts revealed about 100 bilateral pairs of GABA immunoreactive neurons with arborizations in the central complex. Five types of immunostained neurons could be identified through reconstruction of the staining pattern, comparison with individually stained neurons, and double labeling experiments with Neurobiotin-injected neurons. All of these GABA-immunostained neurons are tangential neurons that connect the lateral accessory lobes to distinct layers of the central body. Three types of immunostained neurons (TL2, TL3, TL4) invade the lower division of the central body, and two additional types of neurons (TU1, TU2) have ramifications in layers I and II of the upper division of the central body. Double-labeling experiments with peptide antisera suggest that peptides related to Phe-Met-Arg-Phe-NH2/bovine pancreatic polypeptide and Dip-allatostatin might act as cotransmitters with GABA in TL4 neurons of the lower division and (Dip-allatostatin only) in TU2 neurons of the upper division of the central body. The high conservation in the pattern of GABA immunostaining in all insect species investigated so far suggests that GABA plays an essential role in the basic neural circuitry of the central complex in insects. PMID- 10379834 TI - Microtubule-dependent movement of symbiotic algae and granules in Paramecium bursaria. AB - Paramecia demonstrate rotational cytoplasmic streaming, in which some cytoplasmic granules and organelles, including symbiotic algae, flow in a constant direction. To elucidate the mechanism of this streaming, we examined the effects of cytochalasins (cytochalasin B and D, and dihydrocytochalasin B) and nocodazole, which are reagents affecting microfilament and microtubule networks, respectively, in the cell. In previous reports, paramecia have been compressed with a coverslip to facilitate observation of cytoplasmic streaming. Here we found that the cytoplasmic streaming of paramecia was suppressed by such compression and then observed the process without compression in this work. In the presence of cytochalasins, cytoplasmic streaming was not affected. In contrast, treatment with nocodazole (10 microg/ml) resulted in discontinuation of cytoplasmic streaming in paramecia. Immunofluorescent microscopic observations by confocal microscopy revealed that the number of intracellular microtubules in nocodazole-treated cells was markedly decreased compared to that of controls. Electron microscopic observations confirmed the decrease. These results suggest that cytoplasmic microtubules play an important role in the cytoplasmic streaming of paramecia. PMID- 10379835 TI - Neuronal cytoskeletal alterations evoked by a platelet-activating factor (PAF) analogue. AB - Platelet-activating factor (PAF), a phospholipid signaling molecule found in brain, modulates several neural functions and is implicated in the human developmental brain disorder Miller-Dieker Lissencephaly (MDL). Exposure to PAF, and a non-hydrolyzable analogue, methyl carbamyl PAF (mc-PAF), produces the following rapid, reversible effects upon cultured hippocampal neurites: growth cone collapse, neurite retraction, and neurite varicosity formation. In this study, the cytoskeletal alterations that mediate these shape changes were investigated by comparing the effects of mc-PAF with other cytoskeletal-altering drugs, through the fluorescent labeling of cytoskeletal proteins and mitochondria, and by electron microscopy. Results indicate that rearrangements of microtubules (MTs), F-actin, and mitochondria underlie the neurite shape changes produced by mc-PAF. Evidence for MT alteration was obtained by comparing the effects of mc-PAF with nocodozole and taxol. Exposure to nocodazole, a MT depolymerizing agent, produced growth cone collapse and neurite varicosity formation similar to mc-PAF, whereas pre-incubation of neurites in taxol, a MT stabilizing drug, was effective in blocking mc-PAF-induced neurite effects. Immunofluorescent labeling and EM revealed MT splaying and unbundling within neurite varicosities following mc-PAF treatment. Immunofluorescent labeling also revealed that F-actin shifted from concentration in the growth cone to a diffuse distribution along the neurite shaft following mc-PAF exposure. Fluorescent labeling and EM also revealed retrograde movement and morphological alterations of mitochondria following mc-PAF exposure, resulting in mitochondrial aggregates within neurite varicosities. These cytoskeletal rearrangements may provide insights into the mechanisms by which PAF influences neuronal activity, and could have important implications for the impairment of neuronal motility observed in MDL. PMID- 10379836 TI - Linkage of a nucleolin-related protein and casein kinase II with the detergent stable photoreceptor cytoskeleton. AB - Vertebrate photoreceptors are highly polarized sensory neurons with a complex microtubule and actin-based cytoskeletal organization. In the present study, we have used a detergent-extracted cytokeleton preparation from bovine photoreceptors to test the hypothesis that protein kinases and their substrates co-purify with the photoreceptor cytoskeleton. We incubated the cytoskeletal preparation in the presence of [gamma-32P]ATP. Following SDS-PAGE and autoradiography, we found two principal phosphoproteins with apparent molecular weights of 55 kDa (pp55) and 112 kDa (pp112). We have additionally identified the kinase responsible for phosphorylation of pp112 (and possibly pp55) as a casein kinase II-like enzyme. pp55 was identified as beta-tubulin based on Western blotting and its position on two-dimensional gels. Microsequencing revealed that 16 of the first 17 amino acids of pp112 were identical to human nucleolin, a nuclear protein. Western blotting, mobility in SDS PAGE and in two-dimensional gels, predominant localization within the nucleus, and phosphorylation by a casein kinase II all support the conclusion that pp112 is a nucleolin-related protein. Immunocytochemistry revealed a significant extranuclear pool of nucleolin-immunoreactivity within the cell bodies of photoreceptors. These findings suggest an important extranuclear role for nucleolin or a related protein in photoreceptors. PMID- 10379837 TI - Purification and biochemical characterization of actin from Caenorhabditis elegans: its difference from rabbit muscle actin in the interaction with nematode ADF/cofilin. AB - Biochemical analysis of cytoskeletal proteins of the nematode Caenorhabditis elegans can be combined with a vast resource of genetic information in order to understand the regulation and function of the cytoskeleton in vivo. Here, I report an improved and efficient method to purify actin from wild-type C. elegans and characterization of its biochemical properties. The purified actin was highly pure and free of several known actin-binding proteins. G-actin was polymerized into F-actin in a similar kinetic process to rabbit muscle actin. G-actin interacted with bovine DNase I and inhibited its activity. However, UNC-60B, an isoform of ADF/cofilin in C. elegans, showed a marked depolymerizing activity on C. elegans actin but not on rabbit muscle actin. The results indicate that C. elegans actin shares common biochemical properties with rabbit muscle actin, while actin-binding proteins can interact with C. elegans actin in a distinct manner from rabbit muscle actin. PMID- 10379839 TI - Locomotory waves of Koruga and Deltotrichonympha: flagella wag the cell. AB - We investigated the nature of the locomotory waves of Koruga and Deltotrichonympha, flagellates living symbiotically in the hindgut of the Australian termite Mastotermes darwiniensis. The locomotory waves consist of two components: metachronal waves of flagellar beating and undulations of the cell surface, which propagate synchronously with the same wavelength, frequency, and velocity. We asked, do body waves cause flagellar waves, or vice versa? Using video microscopy and selective inhibitors and drugs, we found that (1) the amplitude of flagellar waves remains constant independent of variations in the amplitude of body waves, (2) flagellar waves can occur in the complete absence of body waves, (3) flagellar waves can induce body waves on swollen regions, (4) inhibition of flagellar beating by dynein inhibitors causes disappearance of body waves, and (5) cytochalasin D induces changes in cell shape but does not inhibit locomotory waves. Therefore, flagellar waves are not produced passively by an active contractile system in the cell cortex; instead, metachronally beating flagella exert waves of pressure that induce passive undulations of a pliant cell surface. These results support Machemer's [1974] theoretical analysis of the data of Cleveland and Cleveland [1966: Arch. Protistenk. 109:39-63], who believed the opposite. PMID- 10379838 TI - Centriole and centrin degeneration during mouse spermiogenesis. AB - Centrosome reduction during mouse spermiogenesis has been studied by immunofluorescent microscopy using anticentrin antibody (20H5) and TEM. Centrin is detected as two spots in round spermatids, corresponding to a pair of centrioles. In elongating spermatids, centrin spots colocalize with the centrioles in the neck region, while the perinuclear ring from which manchette microtubules arise, does not label with the antibody 20H5. The proximal centriole of the elongating spermatids develops a prominent adjunct, which assembles an aster of microtubules. TEM studies after immunogold labeling revealed that centrin is associated with the distal and the proximal centrioles, but not with the adjunct. Centrin labeling in the neck region diminishes after spermiation stage, although it is not completely lost from all testicular sperm. Mature epididymal sperm do not display centrin labeling. Mouse sperm lose both distal and proximal centrioles at maturity. Loss of centrin staining appears to correlate with the degeneration of centrioles during mouse spermiogenesis. PMID- 10379840 TI - Poly(A) mRNA is attached to insect ovarian microtubules in vivo in a nucleotide sensitive manner. AB - In ovarioles of hemipteran insects, RNA passes from anteriorly positioned nurse cells to the chain of developing oocytes via extended nutritive tubes. These intercellular connections may reach several millimeters in length. Each nutritive tube is comprised of many thousands of parallel microtubules. We have extracted microtubule bundles from isolated nutritive tubes of Notonecta glauca and, using hybridization techniques, provide evidence of poly(A) mRNA attachment to microtubules in vivo. We also show this attachment to be nucleotide-sensitive, which is typical of a motor protein-mediated interaction. The pattern of nucleotide sensistivity is indicative of a kinesin motor mechanism. We provide evidence that a kinesin is present in the nutritive tube translocation channels and is a component of the mRNA/microtubule bundles isolated and extracted from them. Our findings are consistent with kinesin-driven transport of mRNA along the nutritive tube microtubules. PMID- 10379841 TI - Assessment of inner dynein arm structure and possible function in ciliary and flagellar axonemes. AB - The construction and assessment of a three-dimensional computer-generated model of inner dynein arms on a 96-nm repeat unit of an axonemal doublet is described. The model is based on published electron micrographs of axonemes from Tetrahymena cilia and eel sperm, which were prepared using several different techniques: negative stain, freeze etch, and thin section. The inner arm structure is represented as three inner dynein arm complexes containing four inner dynein arms (IDAs), three dyads, and one single-headed arm, each capable of bridging the interdoublet gap. The IDA structures in the model have been correlated with the domains containing dynein heavy-chain isoforms mapped by several authors using genetic analyses of Chlamydomonas mutants. The model is consistent with micrographic evidence from axonemes of cilia and flagella from other organisms that led previously to conflicting structural interpretations. In this reconciling interpretation, the different alignments of the IDAs relative to the corresponding outer dynein arms observed in micrographs of differently prepared samples, result from the IDAs being arrested at different stages of their cycles of activity in each preparation. By interpolating between these positions of arrest, cycles of activity are proposed for each of the IDAs during which the arms attach to the neighbouring doublet microtubule and drive it tipwards. PMID- 10379842 TI - A possible role for granulocyte macrophage-colony stimulating factor in modulating teratogen-induced effects. AB - It was already shown that stimulation of the maternal immune system by allogeneic or xenogeneic leukocytes is capable of affecting embryonic responses to teratogenic insults and various cytokines, including granulocyte macrophage colony stimulating factor (GM-CSF), were implicated as mediators of this effect. Therefore, in the present study we tried to assess the ability of GM-CSF to modulate teratogenic activity, along with possible changes in systemic as well as local maternal immune responses, that might be involved in the process. Thus, the percentage of cyclophosphamide (CP)-treated embryos exhibiting limb malformations was shown to decrease significantly following GM-CSF administration. This effect was found to be comparable to that demonstrated by intrauterine leukocytes administration. GM-CSF treatment resulted in a significant enhancement in maternal splenocytes Con-A-induced proliferation, as well as Interleukin-2 (IL-2) and IL-3 production. Examination of leukocyte cell surface antigens expressed by splenocytes revealed no statistically-significant changes in the level of the T lymphoid antigens Thy-1, CD5, CD4, CD8 and CD3, the macrophage antigen Mac-1, and the adhesion molecules LFA-1alpha, LFA-1beta and L-Selectin, following GM-CSF immunostimulation. In parallel, immunohistochemical analysis of the uteroplacental unit revealed Mac-1 and to a lesser extent LFA-1beta-positive cells localized to the myometrium and the placenta in both the control and the GM CSF-treated groups, while no cells expressing Thy-1, CD3, CD4, CD8, or LFA-1alpha could be demonstrated. Our results suggest a possible role for GM-CSF in modulating teratogen-induced effects, a process in which maternal immune responses such as splenocytes proliferation and cytokine production might be involved. PMID- 10379843 TI - Concurrent teratogenic and mutagenic action of 2-methoxyethanol in Drosophila melanogaster larvae resulted in similar phenotypes: close resemblance to directed mutations. AB - Early third instar larvae of wild-type Drosophila melanogaster were transferred to medium supplemented with 2-methoxyethanol (2-ME) or ethylene glycol monomethyl ether. 2-ME produced terata in adult flies as wing notches and duplications of macrochaetae similar to feeding with methoxyacetic acid (MAA), the oxidation product of 2-ME. Larval feeding with 2-ME also affected the fertility of both females and males. 2-ME or more likely its intermediate oxidation product, methoxyacetaldehyde (MAALD), concurrently generated mutations in the premeiotic stages of the oocytes in the early third instar larvae. The mutagenicity of 2-ME has been confirmed in subsequent small scale experiments. The mutation frequency ranged from 4 x 10(-4) to 1 x 10(-2). Although terata were not supposed to be heritable, 1.1 to 8.7% of the affected females produced offspring with phenotypic similarity to the female parent. This phenomenon looked like a classical example of inheritance of an acquired character. The question is addressed why a Notch like phenocopy, generated by larval 2-ME treatment, could bring forth Notch and rudimentary mutants in particular. Administration of 2-ME to larvae, containing the highly active alcohol dehydrogenase variant ADH-71k, exposed the mitotic germ cells and the mitotic somatic cells of the imaginal discs simultaneously to the mutagen MAALD and the teratogen MAA, respectively. The chances for specific gene mutations, though non-adaptive, were likely increased by a feedback mechanism: gene-products that were inhibited or disturbed by the teratogen demanded increased transcription of their encoding genes. Transcribed genes are more susceptible to mutagens. PMID- 10379844 TI - Developmental toxicity of indium in cultured rat embryos. AB - Developmental toxicity of indium was examined using rat embryo culture with reference to toxicokinetics. Rat embryos at day 9.5 of pregnancy were cultured for 48 h under various exposure conditions to indium trichloride. Indium was embryotoxic to cultured rat embryos at concentrations ranging from 25 to 50 microM for 24 h exposure according to the embryonic age, and the exposure concentration was more critical than the exposure time. The embryotoxic concentrations were comparable to the serum concentration at a developmentally toxic dose by intravenous administration in an in vivo experiment. It was considered from these results that the developmental toxicity of indium is a direct effect on the embryo or yolk sac and that weak developmental toxicity of indium by oral administration was due to low exposure concentrations in the embryo. PMID- 10379845 TI - N-demethylation of phenothiazines by lipoxygenase from soybean and human term placenta in the presence of hydrogen peroxide. AB - Several phenothiazine derivatives have been shown to cause reproductive toxicity. The biochemical mechanisms responsible for these effects are not fully understood at present. In this study, we investigated hydrogen peroxide-dependent oxidation of six phenothiazines by purified lipoxygenase from soybean (SLO) and human term placenta (HTPLO). Chlorpromazine was employed as the prototype phenothiazine drug. Chlorpromazine was easily demethylated releasing formaldehyde when incubated at pH 7.0 and 6.5 with SLO or HTPLO, respectively, in the presence of hydrogen peroxide. The reaction was linear with respect to time, exhibited dependence on the amount of enzyme, and the concentration of chlorpromazine and hydrogen peroxide. Under the optimal assay conditions, the estimated Vmax values for chlorpromazine N-demethylation were 139 and 7.2 nmoles/min/mg of SLO and HTPLO, respectively. Collectively, the results suggest an enzymatic nature of the reaction. In the presence of gossypol and NDGA, the classical inhibitors of different lipoxygenases, the formaldehyde production was significantly decreased, as expected. Similar to SLO, the generation of chlorpromazine cation radical, an initial oxidation product with an absorption maximum at 525 nm, was also observed with HTPLO. The radical generation was detectable only under acidic conditions (pH 3.5-4.5). The formaldehyde production was also decreased by BHT and BHA, suggesting a radical nature of the SLO-mediated chlorpromazine N-demethylation. Reduced glutathione, ascorbate, and dithiothreitol suppressed the rate of SLO dependent formaldehyde generation, presumably due to the reduction of the cation radical back to chlorpromazine in a concentration-dependent manner. Besides chlorpromazine, SLO also oxidized promazine, triflupromazine, trifluperazine, trimeprazine, and perphenazine, albeit at different rates, in the presence of hydrogen peroxide. The evidence gathered in this in vitro study suggests that phenothiazines can undergo peroxidative N-demethylation via lipoxygenase pathway. The role of this biochemical mechanism in the in vivo developmental toxicity of phenothiazines remains to be established. PMID- 10379846 TI - Combined prenatal toxicity of 6-mercaptopurine riboside and hydroxyurea in mice. AB - Hydroxyurea (HU) and 6-mercaptopurine riboside (6-MPr) are used as cytostatic chemotherapeutics. Their teratogenic potential in experimental animals has been well known for several decades. Generally, it is assumed that the toxicity of both agents is due to an interference with enzymes of DNA synthesis. In the case of 6-MPr, it was speculated that the teratogenicity in rodents might be paralleled by or even correlated to an incorporation of 6-thioguanine into the DNA of the embryos. In this study, the interaction between these two compounds with regard to teratogenicity in NMRI mice was investigated. Dose-response data of 6-MPr (s.c.-treatment) were published earlier. First, a dose-response study with HU alone (i.p.-treatment) was performed. From these data, the doses for the combination study were derived: HU 250 mg/kg (NOAEL dose) and 6-MPr 16 mg/kg (strongly teratogenic dose). In all experimental groups the substances were administered to the dams once on day 11 of gestation. Combination effects were investigated applying various dosing regimens. In group I treatment was simultaneous, in group II HU was administered 2 h before 6-MPr, in group III 2 h after 6-MPr. The differences in the overall frequency of gross structural abnormalities were moderate. However, when analysing the effects in more detail (single abnormalities), group III exhibited great differences: 1) 6-MPr co treatment increased the frequency of HU effects (skull defects) and 2) HU co treatment decreased the frequency of 6-MPr effects (limb defects) when compared to the findings of the dose-response studies. In addition, the influence of HU on the 6-MPr-induced DNA modification was determined by measuring the incorporation of 6-thioguanine into the DNA of day-11 embryos. As expected, the HU co-treatment corresponding to the group III dosing regimen of the teratogenicity experiment decreased the incorporation rate by ca. 40%. This was in parallel to the decrease in the frequency of 6-MPr effects in the teratogenicity III group. This finding may be considered as a further indication that in the case of 6-MPr, DNA modification is accompanying teratogenicity. PMID- 10379847 TI - Developmental toxicity of the topoisomerase inhibitor, etoposide, in rabbits after intravenous administration. AB - The developmental effect of the topoisomerase inhibitor, etoposide, was investigated in pregnant rabbits given intravenous doses during early organogenesis. Does received 0, 0.25, 0.5, 1, or 2 mg/kg/day on days 7 through 9 of gestation. Fetal parameters were evaluated on day 28 of gestation. Live fetuses were examined for gross, visceral, and skeletal malformations and variations. In addition, telencephalon in embryos 8 h following the final treatment was examined histologically. No change in general condition was observed in any does, but a significant decrease in body weight gain during the pregnancy and enlargement of the liver resulting from marked fatty change were observed in does treated with etoposide at 2 mg/kg/day. Etoposide had neither lethal nor growth retarded effects on embryos/fetuses. However, axial skeletal malformation and extra ribs had a low incidence but were significant in the group treated with etoposide at 2 mg/kg/day, whereas no significant increases in external malformations in term fetuses nor in pyknotic cells in the ventricular zone of telencephalon in embryos were noticed in any etoposide-treated groups. It was concluded that anatomical defects (skeletal malformation or variation) in rabbits were induced by intravenous etoposide treatment during early organogenesis and that they occurred in the presence of maternal toxicity. PMID- 10379848 TI - The breast biopsy paradigm shifts once again. PMID- 10379849 TI - Surgery for melanoma metastatic to the gastrointestinal tract. PMID- 10379850 TI - Cutaneous lymphatic drainage patterns in patients with grossly involved nodal basins. PMID- 10379851 TI - Coil embolism as rescue--the solution to misperfusion. PMID- 10379852 TI - Extent of lumpectomy for breast cancer after diagnosis by stereotactic core versus wire localization biopsy. AB - BACKGROUND: Stereotactic core biopsy of mammographically defined breast abnormalities is an alternative to wire localization biopsy. The purpose of this study was to evaluate the extent of lumpectomy in patients diagnosed by stereotactic core versus wire localization biopsy. METHODS: A total of 67 consecutive patients diagnosed with invasive cancers or ductal carcinoma in situ (DCIS) were retrospectively reviewed. Thirty-four were diagnosed by core biopsy and the remaining 33 by wire localization biopsy. RESULTS: Approximately 65% of patients subsequently had breast-conserving surgical therapy. Seventy-nine percent of patients undergoing wire localization biopsies had positive surgical margins. Achievement of negative surgical margins for lumpectomies performed after wire localization or stereotactic core biopsies was 100% and 89%, respectively, which was not significantly different. However, the total volume of breast tissue removed for breast conservation in patients undergoing lumpectomy after wire localization versus core biopsies was 183 cm3 and 104 cm3, respectively, which was significantly different (P = .003). CONCLUSIONS: Diagnosis by stereotactic core biopsies resulted in less tissue removal to achieve margin-negative lumpectomies for breast conservation. Stereotactic core biopsy is the method of choice for biopsying nonpalpable, suspicious breast lesions. PMID- 10379853 TI - Surgery for melanoma metastatic to the gastrointestinal tract. AB - BACKGROUND: Gastrointestinal (GI) metastasis from melanoma has a dismal prognosis with few long-term survivors. We evaluated the role of operative intervention for melanoma metastases to the GI tract and attempted to identify prognostic factors to improve selection of patients for surgery. METHODS: Between 1977 and 1997, 68 of the 7965 patients with melanoma admitted to Memorial Sloan-Kettering Cancer Center underwent surgical exploration for melanoma metastatic to the GI tract. Characteristics of the primary tumor, regional lymph nodes, and metastatic pattern were reviewed. Data concerning the presenting signs and symptoms, laboratory values, operative findings, extent of surgical resection, recurrence pattern, and survival were analyzed. RESULTS: The most common presenting clinical features included anemia (n = 41; 60%) or abdominal pain (n = 40; 59%). The most frequently involved portion of the GI tract was the small bowel (n = 62; 91%), and the most common operative procedure was small bowel resection (n = 54; 79%). Postoperative mortality and morbidity were 2.9% (n = 2) and 8.8% (n = 6), respectively. Presenting symptoms were relieved in 90% of patients (n = 61). Median survival for all 68 patients following operative intervention was 8.2 months, with 18% survival at 5 years. By multivariate analysis, complete resection rendering the patient free of all identifiable disease (n = 19, median survival 14.9 months, 38% survival at 5 years) and a low preoperative serum lactate dehydrogenase (LDH) (n = 28, median survival 13.6 months, 35% survival at 5 years) were identified as independent favorable prognostic factors for survival. CONCLUSIONS: Operative intervention for melanoma metastatic to the GI tract is recommended for palliative reasons and can be performed with low morbidity and mortality. It is associated with prolonged survival in patients rendered free of all identifiable disease following surgical resection and in those with a low preoperative serum LDH. PMID- 10379854 TI - Cutaneous lymphatic drainage in patients with grossly involved nodal basins. AB - BACKGROUND: The development of lymphatic mapping techniques has facilitated the identification of the sentinel lymph node (SLN), the first node in the regional basin into which cutaneous lymphatics flow from a particular skin area. Previous studies have shown that SLN histology reflects the histology of the entire basin, because melanoma metastases progress in an orderly fashion, involving the SLN before higher nodes in the basin become involved with metastatic disease. It is uncertain whether these orderly cutaneous lymphatic flow patterns are maintained in grossly involved basins. Lymphatic mapping was performed in a population of melanoma patients with clinically palpable lymphadenopathy to address this question. We aimed to determine whether the presence of gross nodal disease in the basin alters lymphatic flow into that basin so that lymphatic mapping techniques are not applicable, and, in patients referred with a grossly involved basin, whether preoperative lymphoscintigraphy should be performed to identify other regional basins at risk for metastases. METHODS: Eight patients presented with grossly palpable disease in the regional basin and underwent preoperative lymphoscintigraphy. All patients with palpable disease and all basins indicated by lymphoscintigraphy to be at risk were dissected. Three patients presented with clinically palpable nodes at the time of diagnosis, and five developed nodal disease on clinical follow-up after undergoing initial wide local excision only. A total of 10 basins in the eight patients were dissected. Of these, eight of the basins had grossly palpable regional nodal disease, and the other two basins were identified by preoperative lymphoscintigraphy as being at risk for metastases. The SLN was identified with intraoperative mapping, harvested, and submitted to pathology. Complete therapeutic lymph node dissections were performed following the SLN harvest in the basins with grossly palpable disease. SLN biopsy alone was performed in the two basins that did not have clinically palpable adenopathy but showed cutaneous lymphatic flow from the scintigram. RESULTS: Sixteen SLNs were harvested from these eight basins with grossly palpable disease, and 14 (87.5%) contained tumor. In each case, one of the SLNs was the grossly palpable node, and in six of the basins (75%) it was the only site of melanoma metastases. An additional 190 higher level, non-SLNs were removed, 32 (16.8%) of which contained microscopic foci of metastatic melanoma (P = .015). The null hypothesis that melanoma nodal metastasis is a random event is rejected. Two patients with trunk melanoma primary sites were identified to have other basins at risk for metastatic disease on lymphoscintigraphy. SLN biopsies were performed in these two patients, and one had microscopic nodal disease in the SLN. CONCLUSIONS: These data support the fact that cutaneous lymphatic drainage patterns are maintained in patients with grossly involved basins, thus buttressing the idea that the SLN is the node most likely to develop metastatic disease. Gross disease in the basin does not significantly alter cutaneous lymphatic flow into the regional basin, as the sentinel lymph node identified under these circumstances is the same as with the grossly involved node. Preoperative lymphoscintigraphy in patients who present with grossly involved nodes in one basin may identify other regional basins with micrometastatic disease and deserves further study in this setting. PMID- 10379856 TI - Subtle differences in quality of life after breast cancer surgery. AB - BACKGROUND: Lumpectomy with axillary dissection (LAD) has taken its place alongside mastectomy (M) as the treatment of choice for stage I and II breast cancer. Its appeal is based on lessening disfigurement and thus improving quality of life. METHODS: We used the SF-36 Health Survey modified with ten questions relevant to breast cancer surgery to evaluate whether quality of life with LAD was better than with mastectomy in women with stage I and II disease. The additional questions addressed satisfaction with intimate relationships and sexuality, and explored impact on the way women dress, use bathing suits, hug people, are comfortable with nudity, and rate their sexual drive and sexual responsiveness. RESULTS: LAD was not associated with statistically significant better quality-of-life scores on any SF-36 questions, except vitality (P = .02). No differences were noted in the areas of intimacy and sexual satisfaction. LAD patients reported significant differences in matters of dress, use of bathing suits, hugging, comfort with nudity, and sexual drive compared to patients undergoing mastectomy. CONCLUSIONS: The SF-36 health survey detected few differences in quality of life measures between patients with LAD and those with mastectomy. However, LAD impacts favorably on the way women dress, on comfort with nudity, and on sexual drive. PMID- 10379855 TI - Transcatheter embolization for the treatment of misperfusion after hepatic artery chemoinfusion pump implantation. AB - BACKGROUND: The use of surgically implanted chemoinfusion pumps for the treatment of hepatic metastases from colorectal carcinoma can be complicated by intra- or extrahepatic misperfusion. This may result in suboptimal tumor exposure to the chemotherapeutic agent and injury to other gastrointestinal organs. Misperfusion can be managed by selective arterial transcatheter embolization. METHODS: Between 1989 and 1996, 16 patients with liver metastases from colorectal carcinoma and with hepatic artery chemoinfusion pump misperfusion were treated using transcatheter coil embolization. Six female and 10 male patients (age range, 34 84 years; median, 51.5 years) were identified by retrospective review of the records of the Department of Interventional Radiology. After pump placement, abnormal liver perfusion scan or methylene blue endoscopy study results prompted angiography with coil embolization. After embolization, the imaging studies were repeated and patients were monitored in the Oncology Clinic. RESULTS: Eight patients exhibited intrahepatic misperfusion (group 1) and eight extrahepatic misperfusion (group 2). Coil embolization was immediately successful in 100% of patients in group 1, with restoration of normal hepatic perfusion, and in 75% in group 2. There were no immediate procedure-related complications. Follow-up periods ranged from 1 to 23 months (median, 13.5 months). Embolization was unsuccessful for two patients (in group 2), who tolerated a modified chemotherapeutic regimen, with follow-up periods of 18.5 and 22 months. CONCLUSIONS: Transcatheter coil embolization is the therapy of choice for the management of hepatic artery chemoinfusion pump misperfusion. It is rapid, effective, and well tolerated by patients and obviates the need for additional surgical intervention. PMID- 10379857 TI - Intraoperative gamma detection of 125I-lanreotide in women with primary breast cancer. AB - BACKGROUND: Somatostatin receptors are present in most human breast cancers. We performed a pilot trial of intraoperative tumor-gamma detection using the radiolabeled somatostatin analog 125I-lanreotide in 13 women with 14 primary breast carcinomas. METHODS: All patients were given 125I-lanreotide intravenously before surgery. Patients underwent lumpectomy, and postresection margins were evaluated with the gamma probe. Axillary dissection specimens were evaluated ex vivo. RESULTS: Seven of 13 women had gamma probe-positive or clinically suspicious margins re-excised at the time of lumpectomy. Four of six probe positive margins were histologically positive, and two of six probe-positive margins were histologically negative; a single clinically suspicious margin was histologically positive. A total of 270 axillary lymph nodes were evaluated ex vivo by gamma probe and histology. McNemar's contingency tests demonstrated a highly statistical correlation between histology and gamma probe counts (P < .0001). CONCLUSIONS: The overall accuracy of nodal evaluation with 125I lanreotide/intraoperative gamma detection was 77%; the negative predictive value of this technique was 97%, however. This technique predicted the presence of tumor in 20% of axillary lymph nodes that were negative by routine histology. This technique appears safe and is able to detect positive tumor resection margins and accurately predict axillary lymph node negativity. Further trials of this technique are required to validate its utility. PMID- 10379858 TI - Markedly elevated levels of vascular endothelial growth factor in malignant ascites. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) is a potent angiogenic factor that also has the ability to increase vascular permeability. Malignant ascites has significant morbidity, but the mechanism of its development is unknown. Because of the permeability-inducing properties of VEGF, we hypothesized that malignant ascites formation is associated with high levels of VEGF. The purpose of our study was to determine the role of VEGF in malignant ascites formation. METHODS: Ascites from 25 patients with gastric (n = 6), colon (n = 7), or ovarian (n = 12) cancers was collected by paracentesis or surgery. VEGF protein levels were determined by enzyme-linked immunosorbent assay. The effect of ascites on endothelial cell permeability was assessed by evaluating propidium iodide uptake by human umbilical vein endothelial cells (HUVECs) exposed to ascites. Neutralizing antibodies to VEGF added to ascites were used to determine the causal effect of VEGF in permeability induction. RESULTS: VEGF protein levels were markedly increased in malignant ascites compared with levels in nonmalignant cirrhotic ascites (controls). VEGF protein levels in ovarian, gastric, and colon cancer ascites were found to be increased 45, 23, and 12 times, respectively, compared with levels in cirrhotic ascites. Malignant ascites from patients with colon and gastric cancer caused an increase in permeability in HUVECs in all cases. Neutralizing VEGF activity in colon cancer ascites decreased in-vitro HUVEC permeability in three of four cases. CONCLUSIONS: VEGF protein levels are markedly elevated in malignant ascites. VEGF may play a role in malignant ascites formation by increasing endothelial cell permeability. PMID- 10379859 TI - Dynamic CT in the preoperative evaluation of patients with gastric cancer: correlation with surgical findings and pathology. AB - BACKGROUND: The use of diagnostic techniques in the preoperative staging of patients with gastric cancer must be better defined. To further clarify which technique is indicated, we applied a new modality of computed tomography (CT) scanning for patients with gastric cancer. METHODS: Dynamic CT of the abdomen using water as oral contrast agent was performed in 30 patients with gastric adenocarcinoma. Patients without evidence of metastatic disease underwent exploratory laparotomy and intraoperative staging. Resectable patients had surgical excision and definitive pathologic staging. RESULTS: Two patients (7%) had metastatic disease by CT and were considered inoperable. The remaining 28 underwent laparotomy. Of these, six (21%) were unresectable and 22 (79%) had surgical resection. Dynamic CT adequately suggested advanced stage disease in four (67%) of the 6 unresectable patients. Wall thickness in dynamic CT correlated with the risk of serosal involvement (P < .001). Both CT and surgery had an accuracy of 64% (P > .05) in predicting pathologic staging. CT overstaged only 4% of cases. CONCLUSIONS: Dynamic CT is a useful modality that can indicate inoperable disease, obviating the need for laparotomy in patients with gastric adenocarcinoma. CT can modify the surgical approach by suggesting unresectable or advanced disease. The low percentage of patients that are overstaged by CT, combined with its similar staging accuracy when compared with laparotomy, support its preoperative use in these patients. PMID- 10379860 TI - Histopathologic criteria for local excision of colorectal cancer: multivariate analysis. AB - BACKGROUND: Local treatment of colorectal cancer, including endoscopic removal of colonic polyps and transanal resection of rectal tumors, has become widely accepted. However, risk factors predicting the presence of lymph node metastasis have not been fully investigated. To determine the criteria for local excision of colorectal cancer, histopathologic factors independently predicting the lymph node metastasis were investigated. METHODS: We performed a retrospective histopathologic study on 335 patients who underwent resection of colorectal cancer and dissection of regional lymph nodes between 1982 and 1996. Features of node-positive tumors (n = 150) were compared with those of node-negative tumors (n = 185), with special reference to the histopathologic findings of the resected tumor. Multivariate analysis was done using the stepwise logistic regression test. RESULTS: Node-positive tumors, when compared with node-negative tumors, were characterized by tumor larger than 6 cm (42% vs. 22%), serosal invasion (88% vs. 56%), lymphatic invasion (32% vs. 5%), venous invasion (9% vs. 2%), and histology other than well-differentiated (66% vs. 29%). Multivariate analysis showed that factors independently associated with lymph node metastasis were serosal invasion, lymphatic invasion, and histologic type. When these three risk factors were negative, lymph node metastasis was rare (5%). When one, two, or three factors were positive, the frequency of lymph node metastasis was 38%, 66%, and 85%, respectively. CONCLUSIONS: In colorectal cancer, factors independently associated with lymph node metastasis are serosal invasion, lymphatic invasion, and histologic type. When these three parameters are favorable, local treatment of colorectal cancer does not require additional lymph node dissection. PMID- 10379861 TI - Myxoid liposarcoma--the frequency and the natural history of nonpulmonary soft tissue metastases. AB - BACKGROUND: Myxoid liposarcomas (ML) make up the major subset of liposarcomas, which in most series represent the second or third most common type of soft tissue sarcoma. The tendency for ML to metastasize to other soft tissues (STM) in preference to lung parenchyma has been previously described; however, the natural history of this tumor's behavior is poorly documented. Our intent was to analyze the natural history of ML and further quantify the incidence of STM, concentrating on their significance in terms of survival. METHODS: We reviewed the experience at the Royal Marsden Hospital over a 10-year period, documenting the clinicopathological behavior of ML, including the frequency of STM. RESULTS: There were 50 patients, with a median follow-up of 43 months. The actuarial 5 year soft tissue metastasis rate was 31%, and the most common sites of STM were the retroperitoneum, abdominal wall, and abdominal cavity. In those 12 patients who had STM there was a median interval of 23 months after original diagnosis to the time the first metastasis became apparent (range, 0-142 months). Median survival following first metastasis was 35 months; 6 of the 12 patients died between 6 and 50 months. Four patients who had STM remain disease free at 15 to 59 months after their first STM. Any round cell component of the ML was associated with a significantly greater chance of metastatic disease (P = .02). In this series, the overall 5-year and 7-year survival rates were 85% and 68%. Patients with STM had an 11 times greater chance of dying than those who did not. CONCLUSIONS: ML usually is an indolent disease, but there is a subset of patients who develop STM and have a significantly worse prognosis. STM can occur years after the initial diagnosis and can be associated with medium-long-term survival after they occur. STM should be managed aggressively because of this. PMID- 10379862 TI - Head and neck cancers associated with Madelung's disease. AB - BACKGROUND: Madelung's disease is a rare lipodystrophy that presents with multiple fatty masses in the neck, trunk, and upper extremities. The fatty accumulation is considered a benign disease, but compression of the aerodigestive tract may occur in long-standing disease. METHODS: Eight Chinese patients with Madelung's disease were reviewed. All were male, aged 48 to 67 years, with a history of disease ranging from 4 to 20 years. Two of the eight patients developed aerodigestive symptoms and were subsequently found to have head and neck cancers. These two patients are described. RESULTS: The possible mechanism that may account for an increase in malignant tumors of the airway in this group of patients is the synergistic effect of smoking and alcohol abuse as risk factors for both Madelung's disease and malignant tumors of the airway. Currently it is recommended that these patients should have their fatty lesions removed surgically. The removal of fat facilitates examination of the neck for signs of cervical lymphadenopathy in malignant disease. CONCLUSIONS: Patients with Madelung's disease should be followed regularly. The development of aerodigestive symptoms should be fully investigated with endoscopy and imaging. The cause of symptoms should not be attributed to fatty compression until a carcinoma of the upper airway has been excluded. PMID- 10379863 TI - Lymph node tumor volumes in patients undergoing sentinel lymph node biopsy for cutaneous melanoma. AB - BACKGROUND: Regional lymph node tumor volumes in patients undergoing sentinel lymph node (SN) biopsy (SNB) for treatment of cutaneous melanoma have not been described. The objectives of this study were to describe the lymph node tumor volumes typically seen in this population and to correlate tumor volumes with tumor thickness and positive SN characteristics. METHODS: Review of a consecutive series of patients with clinically localized cutaneous melanoma who underwent SNB of nonpalpable regional lymph node basins followed by complete lymphadenectomy (LND) was performed. Multiple lymph node sections from positive SNs and nonsentinel nodes (NSNs) in LND specimens were examined microscopically. Individual tumor deposit diameters were measured using an ocular micrometer. Aggregate tumor volumes were calculated for SN and LND specimens. Tumor volumes and SN and LND positivity rates were correlated with tumor thickness, the number of positive SNs, and the presence of multiple SN tumor deposits. RESULTS: SNB procedures were performed for 149 melanomas in 189 regional nodal basins. The mean tumor depth was 2.48 mm. The mean number of SNs/basin was 2.1. Thirty-two of 149 SNB procedures (21.5%) revealed a total of 34 nodal basins with at least one positive SN. The median tumor volume in positive SNs was 4.7 mm3 (range, 0.1-3618 mm3; mean, 209 mm3). The median aggregate tumor volume in positive LND specimens was 4.9 mm3 (range, 0.1-3618 mm3; mean, 224 mm3). Six basins (17.6%) contained at least one positive NSN. The regional node aggregate tumor volume correlated weakly with tumor thickness (Pearson's correlation coefficient = .302, P = .0934). NSN positivity was not predicted by tumor thickness, American Joint Committee on Cancer tumor stage, number of positive SNs, or number of metastatic deposits within SNs. CONCLUSIONS: Most melanoma-positive SNs contain minute tumor volumes. Tumor thickness and patterns of SN metastases may not be predictive of tumor burden or the presence of positive NSNs. PMID- 10379864 TI - Absence of severe systemic toxicity after leakage-controlled isolated limb perfusion with tumor necrosis factor-alpha and melphalan. AB - BACKGROUND: Severe systemic toxicity and hemodynamic changes after isolated limb perfusion (ILP) with tumor necrosis factor-alpha (TNF-alpha) and melphalan, with or without interferon-gamma, have been reported in several series. We studied whether these side effects could be precluded by preventing leakage from the isolated circuit into the systemic circulation. METHODS: Clinical and pharmacokinetic data for 20 consecutive patients with recurrent melanoma of the limbs who were treated by ILP with TNF-alpha (3-4 mg) and melphalan, with or without interferon-gamma, were studied. Leakage rates and TNF-alpha levels were determined during and after ILP and were correlated with systemic toxicity and hemodynamic changes. RESULTS: Only two patients experienced leaks (2% and 13%) during ILP. For 18 patients without leakage, the mean peak systemic TNF-alpha level was 2.8 ng/ml at 10 minutes after ILP. After leakage, the peak systemic TNF alpha levels were 31.9 and 88.3 ng/ml at 5 minutes. Toxicity was mild and consisted mainly of fever (n = 17) and nausea/vomiting (n = 19) during the first day after ILP. Some patients developed tachycardia (n = 6), hypotension (n = 3; responding immediately to fluid challenge), a decrease in the WBC count (n = 3; grade I) or thrombocyte count (n = 11; grade I/II, no hemorrhage or therapeutic intervention), or hepatotoxicity [cytolysis (n = 15; 14 grade I/II and 1 grade IV) or hyperbilirubinemia (n = 7; grade I/II, all resolving spontaneously)]. Patients with tachycardia or hepatotoxicity exhibited significantly higher TNF alpha levels after ILP, compared with other patients. CONCLUSIONS: Systemic toxicity after ILP with TNF-alpha is minimal and does not differ from that after ILP with melphalan alone when leakage is adequately controlled. PMID- 10379865 TI - Genomic changes defining the genesis, progression, and malignancy potential in solid human tumors: a phenotype/genotype correlation. AB - The transition of normal epithelium to invasive carcinoma occurs sequentially. In colorectal and cervical carcinogenesis, this transition is reflected by histomorphologically defined grades of increasing dysplasia that untreated may progress to invasive disease. In an attempt to understand the role of chromosomal aberrations during tumorigenesis we have applied comparative genomic hybridization using DNA extracted from defined stages of colorectal and cervical tumors, from low- and high-grade astrocytic tumors and from diploid and aneuploid breast carcinomas. Genetic instability, as measured by the number of chromosomal copy alterations per case, increases significantly at the transition from precursor lesions to invasive carcinomas and continues to increase with tumor stage. Aggressive tumors have a higher number of copy alterations per case. High level copy number changes (amplifications) become more prevalent in advanced stage disease. Subtractive karyograms of chromosomal gains and losses were used to map tumor stage-specific chromosomal aberrations and clearly showed that nonrandom chromosomal aberrations occur during disease progression. In colorectal and cervical tumors, chromosomal copy number changes were correlated with nuclear DNA content, proliferative activity, expression levels of the tumor suppressor gene TP53, and the cyclin-dependent kinase inhibitor p21/WAF1, as well as the presence of viral genomes. Here we summarize and review the results of this comprehensive phenotype/genotype correlation and discuss the relevance of stage specific chromosomal aberrations with respect to diagnostic applications. PMID- 10379866 TI - Genomic imbalances of 7p and 17q in malignant peripheral nerve sheath tumors are clinically relevant. AB - We investigated 31 malignant peripheral nerve sheath tumors (MPNSTs) from 23 patients by means of comparative genomic hybridization (CGH) in order to study quantitative genomic aberrations of these tumors. Twenty-one of the 23 patients revealed changes, with a mean value of 11 aberrations per sample (range 2-29). The minimal common regions of the most frequent gains were 8q23-q24.1 (12 cases), 5p14 (11 cases), and 6p22-pter, 7p15-p21, 7q32-q35, 8q21.1-q22, 8q24.2-qter, and 17q22-qter (10 cases each). Seventeen high-level amplifications were detected in eight of the 21 samples. In three cases, the high-level amplifications involved 8q24.1-qter, and in two cases each the high-level amplifications involved regions 5p14, 7p14-pter, 8q21.1-q23, and 13q32-q33. The minimal common region of frequent losses was 14q24.3-qter (five cases). The gain of 8q as a single common change in the primary tumor, the recurrence, and the metastasis from the same patient suggests that this aberration is an early change in the tumorigenesis of MPNSTs. Comparable aberrations were observed in separate tumors of the same patients affected by Recklinghausen's disease, indicating a limited number of accidental secondary changes. In sporadic MPNSTs, the most frequent gains were narrowed down predominantly to 5p, 6, 8q, and 20q, whereas in MPNSTs from patients with Recklinghausen's disease, there was most often a gain in 7q, 8q, 15q, and 17q. The occurrence of gain of both 7p15-p21 and 17q22-qter was associated with a statistically significant poor overall survival rate (P = 0.0096). PMID- 10379867 TI - Loss of heterozygosity at 11q23.1 and survival in breast cancer: results of a large European study. Breast Cancer Somatic Genetics Consortium. AB - Among the chromosomal regions commonly undergoing deletions in breast tumors is 11q23.1. The genes that are targets for loss of heterozygosity (LOH) in this region is not yet established. One of the candidate genes located in this region is ATM, responsible for the rare autosomal recessive disorder ataxia telangiectasia (A-T). Interestingly, A-T heterozygotes may have an increased risk of cancer, in particular breast cancer, although this is still controversial. A common assumption has been that the target for the LOH at 11q23.1 in breast carcinoma is the ATM gene, but the area studied has been too large, the density of markers too low, and the number of tumors studied has been too small to draw any firm conclusions. The present study is a multicenter study including 918 breast cancer patients with clinical information and survival data available for most of them. Primary breast tumors were investigated for LOH using a high density of microsatellite markers spanning approximately 6 Mb around the ATM gene. Survival analyses showed that there are most likely one or more candidate genes in a 3-4 Mb region between the markers D11S1819 and D11S927 including the ATM gene. Cancer-specific survival was significantly reduced in patients whose tumors exhibited LOH of markers D11S2179 (within the ATM gene), D11S1778, D11S1294, and D11S1818. The highest survival hazard ratios were 1.8(C11.2-2.8, P = 0.010) and 2.1 (C11.4-3.0, P = 0.0004) for markers D11S2179 and D11S1818, respectively. One or more of these markers are therefore most likely to be located close to or within genes associated with breast cancer survival. PMID- 10379868 TI - Myeloid- and lymphoid-specific breakpoint cluster regions in chromosome band 13q14 in acute leukemia. AB - Abnormalities of chromosome band 13q14 occur in hematologic malignancies of all lineages and at all stages of differentiation. Unlike other chromosomal translocations, which are usually specific for a given lineage, the chromosomal translocation t(12;13)(p12;q14) has been observed in both B-cell and T-cell precursor acute lymphoblastic leukemia (BCP-, TCP-ALL), in differentiated and undifferentiated acute myeloblastic leukemia (AML), and in chronic myeloid leukemia (CML) at progression to blast crisis. The nature of these translocations and their pathologic consequences remain unknown. To begin to define the gene(s) involved on chromosome 13, we have performed fluorescence in situ hybridization (FISH) using a panel of YACs from the region, on a series of 10 cases of acute leukemia with t(12;13)(p12;q14) and 1 case each with "variant" translocations including t(12;13)(q21;q14), t(10;13)(q24;q14) and t(9;13)(p21;q14). In 8/13 cases/cell lines, the 13q14 break fell within a single 1.4 Mb CEPH MegaYAC. This YAC fell immediately telomeric of the forkhead (FKHR) gene, which is disrupted in the t(2;13)(q35;q14) seen in pediatric alveolar rhabdomyosarcoma. Seven of the 8 cases with breaks in this YAC were AML. In 4/13 cases, the 13q14 break fell within a 1.7-Mb YAC located about 3 Mb telomeric of the retinoblastoma (RB1) gene: all 4 cases were ALL. One case of myelodysplastic syndrome exhibited a break within 13q12, adjacent to the BRCA2 gene. These data indicate the presence of myeloid- and lymphoid-specific breakpoint cluster regions within chromosome band 13q14 in acute leukemia. PMID- 10379869 TI - Mapping of the breakpoints on the short arm of chromosome 17 in neoplasms with an i(17q). AB - Isochromosomes are monocentric or dicentric chromosomes with homologous arms that are attached in a reverse configuration as mirror images. With an incidence of 3 4%, the i(17q) represents the most frequent isochromosome in human cancer. It is found in a variety of tumors, particularly in blast crisis of chronic myeloid leukemia (CML-BC), acute myeloid leukemia (AML), non-Hodgkin's lymphoma (NHL), and medulloblastoma (MB), and indicates a poor prognosis. To determine the breakpoints on the molecular genetic level, we analyzed 18 neoplasms (six CML, four AML, one NHL, and seven MB) with an i(17q) and two MB with a pure del(17p) applying fluorescence in situ hybridization (FISH) with yeast artificial chromosome (YAC) clones, P1-artificial chromosome (PAC) clones, and cosmids from a well-characterized contig covering more than 6 Mb of genomic DNA. We identified four different breakpoint cluster regions. One is located close to or within the centromere of chromosome 17 and a second in the Charcot-Marie-Tooth (CMT1A) region at 17(p11.2). A third breakpoint was found telomeric to the CMT1A region. The fourth, most common breakpoint was detected in MB, AML, and in CML-BC specimens and was bordered by two adjacent cosmid clones (clones D14149 and M0140) within the Smith-Magenis syndrome (SMS) region. These results indicate that the low copy number repeat gene clusters which are present in the CMT and SMS regions may be one of the factors for the increased instability that may trigger the formation of an i(17q). PMID- 10379870 TI - Novel method for the production of multiple colour chromosome paints for use in karyotyping by fluorescence in situ hybridisation. AB - The development of 24-colour fluorescence in situ hybridisation (FISH) has led to significant advances in cytogenetic research and offers the potential for automated karyotypic analysis. However, these techniques are not in routine research or clinical use because of limitations in methods of probe preparation. This article presents new probe construction protocols and strategies for multiple-colour karyotyping by chromosome painting, which makes the technique more efficient and may lead to more widespread implementation. We used paints generated by our protocols to demonstrate the presence of a cryptic translocation t(13;11;22) in the paediatric sarcoma cell line RMS 1598. PMID- 10379871 TI - A recurring pattern of chromosomal aberrations in mammary gland tumors of MMTV cmyc transgenic mice. AB - Mice carrying the MMTV-cmyc transgene develop mammary tumors at 9 to 12 months of age. Little is known about karyotypic changes in this model of human breast cancer. We have developed and applied molecular cytogenetic techniques to study chromosomal aberrations that occur in these tumors, namely, comparative genomic hybridization and spectral karyotyping. Cell lines from eight tumors were established and analyzed, four of which carried a heterozygous p53 mutation. All of the tumor cell lines revealed increases in ploidy and/or multiple numerical and structural chromosomal aberrations. No consistent differences were observed between cmyc/p53+/+ and cmyc/p53+/- tumors, suggesting that cmyc induces karyotype instability independent of p53 status. Loss of whole chromosome (Chr) 4 was detected in five of the eight tumors. Parts of Chr 4 are syntenic to human 1p31-p36, a region that is also deleted in human breast carcinomas. Four tumors carried translocations involving the distal portion of Chr 11 (syntenic to human chromosome arm 17q), including two translocations T(X;11), with cytogenetically identical breakpoints. We compare the pattern of chromosomal aberrations with human breast cancers, find similarities in several syntenic regions, and discuss the potential of an interspecies cytogenetic map of chromosomal gains and losses. PMID- 10379872 TI - 1q23 gain is associated with progressive neuroblastoma resistant to aggressive treatment. AB - Neuroblastoma is one of the most common malignant tumors of childhood and is characterized by regressive and progressive disease. Genetic factors that define progression of neuroblastomas are still unknown. We performed comparative genomic hybridization (CGH) on 27 neuroblastomas and dual-color fluorescence in situ hybridization (FISH) to identify genetic aberrations associated with progressive neuroblastoma showing resistance to aggressive treatment. 17q21-q25 gains and MYCN amplification were associated with stage 4 neuroblastomas; however, these genetic aberrations had no significant relation to the progression of stage 4 neuroblastomas. A novel chromosomal gain at 1q21-q25 was found in 8 of 16 cases (50%) of stage 4 neuroblastoma. Gain of 1q21-q25 was observed in all of the progressive cases (8/8), which showed resistance to chemotherapy, including 5 fatal neuroblastomas in stage 4, whereas 1q21-q25 gain was not found in any of the 8 remission cases in stage 4. Survival analysis also showed that 1q21-q25 gain was associated with a poor outcome. High xenotransplantability in nude mice was observed for the tumors with 1q21-q25 gain (4/5; 80%). These data show that 1q21-q25 gain is strongly associated with progression of stage 4 neuroblastoma. Furthermore, by dual-color FISH analysis using cosmid clones, the 1q21-q25 gain was narrowed to increase in DNA copy number on 1q23 in the fatal type of stage 4 neuroblastoma showing this gain. These results suggest that DNA amplification at 1q23 may play a role in the development of progressive neuroblastoma in an advanced stage. PMID- 10379873 TI - Deletion mapping at 12p12-13 in metastatic prostate cancer. AB - The identification of homozygous deletions in malignant tissue is a powerful tool for the localization of tumor suppressor genes. Representational difference analysis (RDA) uses selective hybridization and the polymerase chain reaction (PCR) to isolate regions of chromosomal loss and has facilitated the identification of tumor suppressor genes, such as BRCA2 and PTEN. We have recently identified a 1-5-cM homozygous deletion on 12p12-13 in a prostate cancer xenograft and found that 47% of patients who died of prostate carcinoma demonstrate focal loss of heterozygosity (LOH) in this region in metastatic deposits. We have now characterized the region of interest by assembling a yeast artificial chromosome (YAC) contig spanning the homozygous deletion and identifying which known genes and expressed sequence tags (EST) lie within the homozygous deletion. A rib metastasis was harvested at autopsy and placed subcutaneously in a male SCID mouse. Genomic DNA from this xenograft and from the patient's normal renal tissue was extracted. Multiplex PCR, with the xenograft and normal DNA used as template, was performed using primers for loci on the Whitehead contig 12.1 believed to be near our region of interest. We found that our deletion lay in a 1-2-Mb interval between WI-664 and D12S358. We then used the same primers to construct a YAC contig across the homozygous deletion. PCR amplification of YAC DNA, using primers for the genomic sequences of known genes and ESTs reported to lie on 12p12-13, was used to identify candidate genes that lay within the deletion. Duplex PCR, with control primers known not to be deleted in the xenograft, was used to confirm that both the CDKN1B and ETV6 genes were homozygously deleted in the xenograft. Mutations in either or both of these genes may play an important role in metastatic prostate carcinoma. PMID- 10379874 TI - Identification and mapping of novel tumor suppressor loci on 6p in diffuse large B-cell non-Hodgkin's lymphoma. AB - Allelic deletions have been thought to be indicators of the presence of tumor suppressor genes (TSGs). As indicated by this allelotype study using 39 highly informative microsatellite markers distributed among all autosomal chromosomes, frequent loss of heterozygosity (LOH) has been found at 6p in B-cell non Hodgkin's lymphoma. To identify the commonly deleted regions (CDRs), we performed fine deletion mapping using 26 highly polymorphic microsatellite markers on 6p. The most frequent LOH occurred at D651721, where 9 of 18 of the informative cases (50%) had allelic losses. Seventeen of 32 cases (53%) exhibited LOH at least at one locus on 6p. Ten of these 17 cases showed interstitial deletions, and their LOH patterns indicated two CDRs on 6p; one between D6S1721 and D6S260 (at 6p23 24), and the other between D6S265 and D6S291 (at 6p21). The genetic distance of both CDRs was 6 cM. The CDKN1A (p21) gene is reported to be located within the interval of the CDR at 6p21, but no mutation of the gene was found in these 32 patients. These data suggested that these two loci might harbor novel putative TSGs responsible for the pathogenesis of malignant lymphoma. We have constructed a contig of yeast artificial chromosome (YAC) clones spanning the most frequent CDR at 6p23-24. This YAC contig can be used for fine physical mapping of the region and cloning of candidate TSGs. PMID- 10379875 TI - Identification of a 1-cM region of common deletion on 4q35 associated with progression of hepatocellular carcinoma. AB - To identify the location of one or more of the putative tumor suppressor genes (TSG) on chromosome arm 4q that may be involved in hepatocellular carcinoma (HCC), we examined 96 primary HCCs for their patterns of allelic loss at 39 microsatellite marker loci distributed along this chromosome arm. Allelic loss at one or more loci was observed in 71 (74%) HCCs. Detailed deletion mapping identified two distinct commonly deleted regions; one was located within a 1-cM interval flanked by D4S1534 and D4S2929 at 4q21-22, the other in the 1-cM interval flanked by D4S2921 and D4S2930 at 4q35. Of the tumors for which clinical data were available, allelic loss at 4q35 was more frequent in poorly or moderately differentiated tumors than in well-differentiated tumors (3/15, 20%, vs. 14/21, 67%, P = 0.008); in tumors larger than 2 cm in size (2/11, 18%, vs. 34/62, 55%, P = 0.046); and in tumors that arose from liver cirrhosis as opposed to HCCs arising from chronic hepatitis (25/42, 60%, vs. 9/27, 33%, P = 0.048). The association of allelic losses on 4q35 with larger tumor size and aggressive histological type implies that loss or inactivation of TSG located within the 1 cM interval of 4q35 identified here contribute to progression of HCCs. PMID- 10379876 TI - Chromosome abnormalities in ovarian adenocarcinoma: I. Nonrandom chromosome abnormalities from 244 cases. AB - Cytogenetics provides important insights into the molecular pathogenesis of human cancers. Although extensive data exist on recurring cytogenetic abnormalities in hematologic cancers, data on individual solid tumor types remain limited. Previous studies of ovarian carcinoma indicated the presence of multiple, complex clonal chromosome abnormalities. Cytogenetics remains one of a few techniques capable of detecting these multiple, simultaneously occurring genetic abnormalities. We describe cytogenetic abnormalities from a series of 244 primary ovarian cancer specimens referred to a single institution. A total of 201/244 cases had fully characterized clonal chromosome abnormalities, of which 134 showed clonal chromosome breakpoints. We used a novel statistical technique to detect nonrandom chromosome breakpoints at the level of chromosome regions. Nonrandom occurrence of chromosome breakpoints was detected at regions 1p1*, 1q1*, 1p2*, 1q2*, 1p3*, 1q3, 3p1*, 1q4*, 6q1*, 6p2, 6q2, 7p1*, 7q1, 7p2*, 11p1*, 11q1, 11q2*, 12p1, 12q2*, 13p1, and 19q1. Simultaneous occurrence of multiple abnormalities was common. However, 120/134 cases had breakpoints at one or more of 13 commonly involved regions (*), suggesting a hierarchy of genetic abnormalities. Among clinical and tumor variables that predict patient survival, tumor grade was significantly associated with the presence of chromosome breakpoints. In additional studies, we show that nonrandom chromosome abnormalities are associated with impaired survival in ovarian cancer and that specific, nonrandomly involved chromosome regions retain significant effects on survival when analyses are controlled for important clinical variables. Additional specific chromosome abnormalities in this series are described, including chromosome gains and losses in near-diploid cases and homogeneously staining regions. These results suggest that recurring, nonrandom chromosome abnormalities are important in the pathogenesis and/or progression of ovarian cancers, and target areas of the genome for molecular genetic studies. PMID- 10379877 TI - Universal linkage system: an improved method for labeling archival DNA for comparative genomic hybridization. AB - Comparative genomic hybridization (CGH) has become a powerful technique for studying gains and losses of DNA sequences in solid tumors. Importantly, DNA derived from archival tumor tissue is also applicable in CGH analysis. However, DNA isolated from routinely processed, formalin-fixed, paraffin-embedded tissue is often degraded, with the bulk of DNA showing fragment sizes of only 400-750 bp. Enzymatic labeling of archival DNA by standard nick translation (NT) decreases DNA size even further, until it becomes too small for CGH (<300 bp). This study presents application in CGH of a commercially available, non-enzymatic labeling method, called Universal Linkage System (ULS), that leaves the DNA fragment size intact. To compare the effect of chemical labeling of archival DNA by ULS vs. enzymatic by NT on the quality of CGH, DNA derived from 16 tumors was labeled by both ULS and NT. In those cases (n = 8), in which the bulk of DNA had a fragment size of 400-1,000 bp, CGH was successful with ULS-labeled probes, but not with NT-labeled probes. In the DNA samples (n = 6) with a fragment size > 1 kb, the intensity of CGH signals was comparable for both ULS- and NT-labeled probes, but CGH with ULS-labeled samples showed a high, speckled, background, which seriously hampered image analysis. In the remaining two cases, which had evenly distributed DNA fragment sizes (range 250-5,000 bp), CGH was successful with both labeling methods. Using DNA fragment size < 1 kb as a selection criterion for ULS labeling, we were able to obtain good quality CGH of a large panel (n = 77) of a variety of archival solid tumors. We conclude that ULS is an excellent labeling method for performing CGH on small-fragment-sized DNA. PMID- 10379878 TI - Molecular mysteries of polycystic ovary syndrome. PMID- 10379879 TI - Regulation of angiogenic growth factors in the female reproductive tract by estrogens and progestins. AB - Proper regulation of angiogenesis and vascular permeability is essential for the physiological functioning of the female reproductive tract, and major health problems in women, such as dysfunctional uterine bleeding, endometriosis, and uterine cancer, involve a vascular component. There is a large body of literature that describes the effects of sex steroids on the vasculature of the reproductive tract, but far less is known about the molecular mechanisms that regulate these important actions. We hope that this minireview will help emphasize the need for mechanistic studies in this area to improve treatment and prevention of these major health problems in women. Specifically, we believe it will be important to 1) define the exact roles of FGF, VEGF, and other factors in physiological and pathological events in the reproductive tract and the cell types and receptors involved; 2) identify estrogen and progesterone receptor subtypes, the DNA elements, nuclear protein factors, and signaling pathways that mediate regulation of these genes by sex steroids; 3) elucidate any mechanisms of cross-talk between sex steroids and other regulatory factors in the overall regulation of FGF, VEGF, and other angiogenic/permeability factors; and 4) eventually understand how genetic polymorphisms of key regulatory elements affect angiogenesis and the regulation of vascular function in the female reproductive tract. PMID- 10379880 TI - Fertility and infertility: genetic contributions from the hypothalamic-pituitary gonadal axis. PMID- 10379881 TI - The actions and interactions of sex steroids and growth factors/cytokines on the skeleton. PMID- 10379883 TI - The placenta and the prolactin family of hormones: regulation of the physiology of pregnancy. PMID- 10379882 TI - Hypothesis: Progesterone primes breast cancer cells for cross-talk with proliferative or antiproliferative signals. AB - In the breast, data from numerous laboratories suggest that cross-talk exists between PR and growth factor and cytokine signaling pathways at multiple levels (Fig. 4). At the cell surface (level 1), progestins up-regulate growth factor and cytokine receptors. We have expanded this observation by examining the effects of progestins in the cytoplasm (level 2) where progestins regulate several intracellular effectors by increasing the levels and altering the subcellular compartmentalization of Stat5, increasing the association of Stat5 with phosphotyrosine-containing proteins and tyrosine phosphorylation of JAK2, Cbl, and Shc, and potentiating EGF-stimulated p42/p44 MAPKs, p38 MAP kinase, and JNK activities. Together, these events lead to sensitization of downstream signaling pathways to the actions of locally acting secondary factors. Finally, inside the nucleus (level 3), agonist-occupied PR synergize with nuclear transcription factors that are growth-factor regulated, to control the activity of key genes involved in breast cell fate (Figs. 1 and 4). We speculate that after progesterone treatment, orchestrated combinations of steroid hormones and growth factors or cytokines can fine tune the timing and degree of expression of a subset of genes that determine whether progestin-primed cells undergo proliferation, differentiation, or programmed cell death. The paradoxical effects of progesterone have presented a longstanding conundrum to the scientist and clinician. Why are physiological levels of progesterone proliferative in the breast but antiproliferative and protective in the uterus? If progesterone is proliferative in the breast, why is high-dose progestin therapy successful in treating breast cancer? Our intent here has been to open a dialogue addressing these questions. Our data and that of others are beginning to show that one cannot approach the question of progestin actions in isolation. Other important regulatory proteins, whose expression may vary in tissue-specific ways, work in concert with progesterone to decide cell fate. The timing and dose of progesterone may also influence the biological response. Since progestins are widely used in oral contraception, in hormone replacement therapy, and in cancer treatments, it is becoming critically important that the subtleties of their mechanisms of action be clearly understood. PMID- 10379884 TI - Targeted expression of Bcl-2 in mouse oocytes inhibits ovarian follicle atresia and prevents spontaneous and chemotherapy-induced oocyte apoptosis in vitro. AB - Members of the Bcl-2 family serve as central checkpoints for cell death regulation, and overexpression of Bcl-2 is known to inhibit apoptosis in many cell types. To determine whether targeted expression of Bcl-2 could be used to protect female germ cells from apoptosis, we generated transgenic mice expressing fully functional human Bcl-2 protein only in oocytes. Transgenic mice were produced using a previously characterized 480-bp fragment of the mouse zona pellucida protein-3 (ZP3) gene 5'-flanking region to direct oocyte-specific expression of a human bcl-2 complementary DNA. Immunohistochemical analyses using a human Bcl-2-specific antibody showed that transgene expression was restricted to growing oocytes and was not observed in the surrounding ovarian somatic cells or in any other nonovarian tissues. Histomorphometric analyses revealed that ovaries collected from transgenic female mice possessed significantly fewer atretic small preantral follicles compared with wild-type sisters, resulting in a larger population of healthy maturing follicles per ovary. However, the number of oocytes ovulated in response to exogenous gonadotropin priming and the number of pups per litter were not significantly different among wild-type vs. transgenic female mice. Nonetheless, oocytes obtained from transgenic mice and cultured in vitro were found to be resistant to spontaneous and anticancer drug-induced apoptosis. We conclude that targeted expression of Bcl-2 only in oocytes can be achieved as a means to convey resistance of the female germ line to naturally occurring and chemotherapy-induced apoptosis. PMID- 10379885 TI - Transgenic models to study gonadotropin function: the role of follicle stimulating hormone in gonadal growth and tumorigenesis. AB - The role of FSH in gonadal tumorigenesis and, in particular, in human ovarian cancer has been debated. It is also unclear what role the elevated FSH levels in the inhibin-deficient mouse play in the gonadal tumorigenesis. To directly assess the role of FSH in gonadal growth, differentiation, and gonadal tumorigenesis, we have generated both gain-of-function and loss-of-function transgenic mutant mice. In the gain-of-function model, we have generated transgenic mice that ectopically overexpress human FSH from multiple tissues using a mouse metallothionein-1 promoter, achieving levels far exceeding those seen in postmenopausal women. Male transgenic mice are infertile despite normal testicular development and demonstrate enlarged seminal vesicles secondary to elevated serum testosterone levels. Female transgenic mice develop highly hemorrhagic and cystic ovaries, have elevated serum estradiol and progesterone levels, and are infertile, mimicking the features of human ovarian hyperstimulation and polycystic ovarian syndromes. Furthermore, the female transgenic mice develop enlarged and cystic kidneys and die between 6-13 weeks as a result of urinary bladder obstruction. In a complementary loss-of-function approach, we have generated double-homozygous mutant mice that lack both inhibin and FSH by a genetic intercross. In contrast to male mice lacking inhibin alone, 95% of which die of a cancer cachexia-like syndrome by 12 weeks of age, only 30% of the double-mutant male mice lacking both FSH and inhibin die by 1 yr of age. The remaining double-mutant male mice develop slow-growing and less hemorrhagic testicular tumors, which are noted after 12 weeks of age, and have minimal cachexia. Similarly, the double-mutant female mice develop slow-growing, less hemorrhagic ovarian tumors, and 70% of these mice live beyond 17 weeks. The double-mutant mice demonstrate minimal cachexia in contrast to female mice lacking only inhibin, which develop highly hemorrhagic ovarian tumors, leading to cachexia and death by 17 weeks of age in 95% of the cases. The milder cachexia-like symptoms of the inhibin and FSH double-mutant mice are correlated with low levels of serum estradiol and activin A and reduced levels of aromatase mRNA in the gonadal tumors. Based on these and our previous genetic analyses, we conclude that elevated FSH levels do not directly cause gonadal tumors. However, these results suggest FSH is an important trophic modifier factor for gonadal tumorigenesis in inhibin-deficient mice. PMID- 10379886 TI - Role of G protein-coupled receptor kinases on the agonist-induced phosphorylation and internalization of the follitropin receptor. AB - The experiments presented herein were designed to identify members of the G protein-coupled receptor kinase (GRK) family that participate in the agonist induced phosphorylation and internalization of the rat FSH receptor (rFSHR). Western blots of human kidney 293 cells (the cell line used in transfection experiments) and MSC-1 cells (a cell line derived from Sertoli cells that displays many of the differentiated functions of their normal counterparts) reveal the presence of GRK2 and GRK6 in both cell lines as well as GRK4 in MSC-1 cells. Cotransfection of 293 cells with the rFSHR and GRK2, GRK4alpha, or GRK6 resulted in an increase in the agonist-induced phosphorylation of the rFSHR. Cotransfections of the rFSHR with GRKs or arrestin-3 enhanced the agonist-induced internalization of the rFHSR, and combinations of GRKs and arrestin-3 were more effective than the individual components. To characterize the involvement of endogenous GRKs on phosphorylation and internalization, we inhibited endogenous GRK2 by overexpression of a kinase-deficient mutant of GRK2 or G alpha t, a scavenger of G betagamma. We also inhibited endogenous GRK6 by overexpression of a kinase-deficient mutant of GKR6. All three constructs were effective inhibitors of phosphorylation, but only the kinase-deficient mutant of GRK2 and G alpha t inhibited internalization. The inhibition of internalization induced by these two constructs was less pronounced than that induced by a dominant-negative mutant of the nonvisual arrrestins, however. The finding that inhibitors of GRK2 and GRK6 impair phosphorylation, but only the inhibitors of GRK2 impair internalization, suggests that different GRKs have differential effects on receptor internalization. PMID- 10379887 TI - Human placental TEF-5 transactivates the human chorionic somatomammotropin gene enhancer. AB - Human chorionic somatomammotropin (hCS) gene expression in the placenta is controlled by an enhancer (CSEn) containing SV40-related GT-IIC and SphI/SphII enhansons. These enhancers are controlled by members of the transcription enhancer factor-1 (TEF-1) family. Recently TEF-5, whose mRNA is abundant in placenta, was shown to bind cooperatively to a unique, tandemly repeated element in CSEn2, suggesting that TEF-5 regulates CSEn activity. However, expression of TEF-5 using a cDNA lacking the 5'-untranslated region and containing a modified translation initiation site was not accompanied by CSEn activation. Using nested, degenerate PCR primers corresponding to conserved TEF domains, several novel TEF 1-related cDNAs have been cloned from a human placental cDNA library. The open reading frame of one 3033-bp clone was identical to TEF-5 and contained 300- and 1423-bp 5'- and 3'-untranslated regions, respectively. The in vitro generated approximately 53-kDa TEF-5 polypeptide binds specifically to GT-IIC and SphI/SphII oligonucleotides. Overexpression of TEF-5 in BeWo cells using the intact 3033-bp cDNA transactivates the hCS and SV40 enhancers and artificial enhancers comprised of tandemly repeated GT-IIC enhansons, but not OCT enhansons. The data demonstrate that TEF-5 is a transactivator that is likely involved in the transactivation of CSEn enhancer function. Further, the data suggest that elements within the untranslated regions, initiation site, or both control TEF-5 expression in ways that influence its transactivation function. PMID- 10379888 TI - Influence of a species-specific extracellular amino acid on expression and function of the human gonadotropin-releasing hormone receptor. AB - The mammalian GnRH receptor is an atypical G protein-coupled receptor which lacks the C-terminal cytoplasmic tail that is present in all other seven-transmembrane domain receptors. The mouse and rat GnRH receptors contain 327 amino acids, whereas human, sheep, and bovine receptors have an additional residue in the second extracellular loop at position 191. Another notable species difference is that human receptors undergo agonist-induced internalization much more rapidly than the mouse receptor. In this report, the role of the additional amino acid (Lys191) in GnRH receptor function was studied in transiently expressed mutant and wild-type human and mouse GnRH receptors. Deletion of Lys191 from the human GnRH receptor caused a 4-fold increase in receptor expression in COS-1 and HEK 293 cells and a modest increase in binding affinity. The magnitude of the agonist induced inositol phosphate response mediated by the deltaK191 human receptor was similar to that of the wild-type receptor, but the EC50 was decreased by about 5 fold. In addition, the rate of internalization of the deltaK191 human receptor was significantly reduced and was similar to that of the mouse receptor. In contrast to these effects of deletion of Lys191, its replacement by Arg, Glu, Gln, or Ala caused no significant change in receptor expression or function. These findings demonstrate that a specific residue in the extracellular region of the human GnRH receptor is a significant determinant of receptor expression, agonist-induced activation, and internalization. PMID- 10379890 TI - Hormone-dependent interaction between the amino- and carboxyl-terminal domains of progesterone receptor in vitro and in vivo. AB - Full transcriptional activation by steroid hormone receptors requires functional synergy between two transcriptional activation domains (AF) located in the amino (AF-1) and carboxyl (AF-2) terminal regions. One possible mechanism for achieving this functional synergy is a physical intramolecular association between amino (N ) and carboxyl (C-) domains of the receptor. Human progesterone receptor (PR) is expressed in two forms that have distinct functional activities: full-length PR-B and the amino-terminally truncated PR-A. PR-B is generally a stronger activator than PR-A, whereas under certain conditions PR-A can act as a repressor in trans of other steroid receptors. We have analyzed whether separately expressed N- (PR A and PR-B) and C-domains [hinge plus ligand-binding domain (hLBD)] of PR can functionally interact within cells by mammalian two-hybrid assay and whether this involves direct protein contact as determined in vitro with purified expressed domains of PR. A hormone agonist-dependent interaction between N-domains and the hLBD was observed functionally by mammalian two-hybrid assay and by direct protein-protein interaction assay in vitro. With both experimental approaches, N C domain interactions were not induced by the progestin antagonist RU486. However, in the presence of the progestin agonist R5020, the N-domain of PR-B interacted more efficiently with the hLBD than the N-domain of PR-A. Coexpression of steroid receptor coactivator-1 (SRC-1) and the CREB binding protein (CBP), enhanced functional interaction between N- and C-domains by mammalian two-hybrid assay. However, addition of SRC-1 and CBP in vitro had no influence on direct interaction between purified N- and C-domains. These results suggest that the interaction between N- and C-domains of PR is direct and requires a hormone agonist-induced conformational change in the LBD that is not allowed by antagonists. Additionally, coactivators are not required for physical association between the N- and C-domains but are capable of enhancing a functionally productive interaction. In addition, the more efficient interaction of the hLBD with the N-domain of PR-B, compared with that of PR-A, suggests that distinct interactions between N- and C-terminal regions contribute to functional differences between PR-A and PR-B. PMID- 10379889 TI - Titration by estrogen receptor activation function-2 of targets that are downstream from coactivators. AB - Cross-interference (squelching) among nuclear receptors has been proposed to reflect the titration of coactivators that bind the receptors in a hormone dependent manner. We have tested whether the coactivators are the only target titrated during squelching of one receptor by another, or whether proteins needed for coactivator function are titrated as well. That the coactivators are indeed one target of squelching is apparent. The isolated ligand-binding domain of the estrogen receptor (ER-LBD) squelches transcriptional activation by the thyroid hormone receptor (TR) only when the LBD is bound to ligands that promote coactivator interactions and only when regions of the LBD that promote coactivator interactions are undisturbed. Furthermore, the ER-LBD and the TR compete in vitro for the related p160 coactivators, SRC1a and GRIP1 (glucocorticoid receptor interacting protein 1), or the putative corepressor, RIP140. Finally TR action becomes more potent when coactivator levels are raised. Nonetheless, supplying excess SRC1a or GRIP1 does not abolish squelching by the ER. In fact, squelching becomes even more severe when coactivators are abundant. Supplying combinations of coactivators from the p160 class and the CREB-binding protein (CBP)/p300 class makes squelching most severe. Elevated RIP140 inhibits TR action, but also protects the residual TR action from squelching by the ER LBD. We conclude that ER-LBD squelches TR both by titrating p160-CBP coactivators and additionally by cooperating with the coactivators to titrate a second factor. The second factor would be needed by the TR for coactivator-mediated transcriptional stimulation. PMID- 10379891 TI - Ribozyme-mediated cleavage of the estrogen receptor messenger RNA and inhibition of receptor function in target cells. AB - Estrogen receptor (ER) functions as a ligand-activated transcription factor for estrogen-regulated genes. Because of the critical role of the ER in the proliferation of certain estrogen-dependent cancer cell types such as the mammary tumor, inhibitors of estrogen action at the level of receptor function are of major clinical interest. Here we describe developments of two ribozymes that can selectively degrade the human ER mRNA and inhibit trans-activation of an artificial promoter containing the estrogen response element. Two ribozymes, designated RZ-1 and RZ-2, cleave the human ER alpha mRNA at nucleotide positions +956 and +889, respectively. These cleavage sites lie within the coding sequence for the DNA-binding domain of the receptor protein. Both RZ-1 and RZ-2 were also effective in inhibiting the progression of quiescent MCF-7 breast cancer cells to the S phase of the cell cycle after their exposure to 17beta-estradiol (10(-9) M). These results provide a new avenue for inhibition of estrogen action by selective mRNA degradation with its potential therapeutic application through targeted gene delivery vectors. PMID- 10379892 TI - Regulation of estrogen receptor activation of the prolactin enhancer/promoter by antagonistic activation function-2-interacting proteins. AB - Transcriptional responses to estrogens are controlled by the cell- and gene specific interactions of the nuclear estrogen receptor (ER) with cofactors and other transcription factors. The pituitary-specific PRL enhancer/promoter is regulated by estrogens only when it is bound by both ER and the pituitary specific transcription factor, Pit-1. Cooperative ER/Pit-1 activation of the dormant PRL enhancer/promoter in pituitary progenitor cells requires the estrogen dependent activation function-2 (AF-2) of ER, but is inhibited by one AF-2 interacting cofactor, RIP140. Here, the complex actions of RIP140 and other AF-2 interacting proteins at the PRL enhancer/promoter were shown to operate via ER itself. RIP140 inhibition of ER/Pit-1 activation in the absence of AF-1 and RIP140 inhibition of both ER alpha and ER beta cooperative activation with Pit-1 suggested a conserved ER site for RIP140 action, possibly AF-2. Coexpression of other AF-2-interacting proteins, including the p160 factors, steroid receptor coactivator-1a (SRC-1a) and glucocorticoid receptor interacting protein-1 (GRIP1), had negligible effects on ER alpha/Pit-1 cooperative activation, but partially relieved RIP140 inhibition. Relief of RIP140 inhibition required the AF 2-binding, LXXLL motifs in SRC-1a and GRIP1. An ER AF-2 mutant that selectively blocked ER interaction with p160s, but not RIP140, still cooperated with Pit-1 and was inhibited by RIP140, but was not relieved by SRC-1a or GRIP1 expression. Thus, SRC-1a and GRIP1 binding to AF-2 counteracted the inhibition of ER/Pit-1 activation by another AF-2-interacting protein, RIP140. Complex, sometimes antagonistic, actions of different classes of AF-2-interacting proteins may play an important role in the cell- and gene-specific estrogen regulation of PRL and other genes. PMID- 10379893 TI - Augmented androgen production is a stable steroidogenic phenotype of propagated theca cells from polycystic ovaries. AB - To test the hypothesis that the hyperandrogenemia associated with polycystic ovary syndrome (PCOS) results from an intrinsic abnormality in ovarian theca cell steroidogenesis, we examined steroid hormone production, steroidogenic enzyme activity, and mRNA expression in normal and PCOS theca cells propagated in long term culture. Progesterone (P4), 17alpha-hydroxyprogesterone (17OHP4), and testosterone (T) production per cell were markedly increased in PCOS theca cell cultures. Moreover, basal and forskolin-stimulated pregnenolone, P4, and dehydroepiandrosterone metabolism were increased dramatically in PCOS theca cells. PCOS theca cells were capable of substantial metabolism of precursors into T, reflecting expression of an androgenic 17beta-hydroxysteroid dehydrogenase. Forskolin-stimulated cholesterol side chain cleavage enzyme (CYP11A) and 17alpha hydroxylase/17,20-desmolase (CYP17) expression were augmented in PCOS theca cells compared with normal cells, whereas no differences were found in steroidogenic acute regulatory protein mRNA expression. Collectively, these observations establish that increased CYP11A and CYP17 mRNA expression, as well as increased CYP17, 3beta-hydroxysteroid dehydrogenase, and 17beta-hydroxysteroid dehydrogenase enzyme activity per theca cell, and consequently increased production of P4, 17OHP4, and T, are stable properties of PCOS theca cells. These findings are consistent with the notion that there is an intrinsic alteration in the steroidogenic activity of PCOS thecal cells that encompasses multiple steps in the biosynthetic pathway. PMID- 10379894 TI - An explanation for observed estrogen receptor binding to single-stranded estrogen responsive element DNA. AB - Estrogen-inducible genes contain an enhancer called the estrogen response element (ERE), a double-stranded inverted repeat. The estrogen receptor (ER) is generally thought to bind to the double-stranded ERE. However, some reports provide evidence that an ER homodimer can bind a single strand of the ERE and suggest that single-stranded ERE binding is the preferred binding mode for ER. Since these two models describe quite different mechanisms of receptor action, we have attempted to reconcile the observations. Analyzing DNA structure by nuclease sensitivity, we found that two identical molecules of a single strand of DNA containing the ERE sequence can partially anneal in an antiparallel manner. Bimolecular annealing produces double-stranded inverted repeats, with adjacent unannealed tails. The amount of annealing correlates exactly with the ability of ER to bind bimolecular EREs. Either strand of an ERE could anneal to itself in a way that would bind ER. We conclude that ER binds only the annealed double stranded ERE both in vitro and in vivo. PMID- 10379896 TI - The parturition defect in steroid 5alpha-reductase type 1 knockout mice is due to impaired cervical ripening. AB - Successful delivery of the fetus (parturition) depends on coordinate interactions between the uterus and cervix. A majority (70%) of mice deficient in the type 1 isozyme of steroid 5alpha-reductase fail to deliver their young at term and thus manifest a parturition defect. Using in vitro and in vivo measurements we show here that rhythmic contractions of the uterus occur normally in these mutant mice at the end of gestation. In contrast, the cervix of the mutant animal fails to ripen at term as judged by biomechanical, histological, and endocrinological assays. Impaired metabolism of progesterone in the cervix of the mutant mice in late gestation leads to an accumulation of this steroid in the tissue. We conclude that a failure of cervical ripening underlies the parturition defect in mice lacking steroid 5alpha-reductase type 1 and that this enzyme normally plays an essential role in cervical progesterone catabolism at the end of pregnancy. PMID- 10379895 TI - Adenovirus-mediated delivery of a dominant negative estrogen receptor gene abrogates estrogen-stimulated gene expression and breast cancer cell proliferation. AB - Dominant negative estrogen receptors are transcriptionally inactive, altered forms of the estrogen receptor (ER) that can dimerize with the ER and have the potential to inactivate the biological functions of this receptor. Here, we provide the first report that adenoviral delivery of a dominant negative ER to ER positive breast cancer cells is able to effectively suppress estrogen-stimulated cell proliferation and the hormonal induction of endogenous genes. We constructed recombinant adenoviral vectors expressing a dominant negative ER (S554 fs, Ad-fs) or, for comparison, antisense ER (Ad-AS), or the sense wild-type ER (Ad-WT). Expression of the dominant negative ER or antisense ER, but not wild-type ER, blocked estradiol stimulation of the estrogen-responsive genes pS2 and c-myc. The dominant negative ER also fully abolished the estradiol-induced increase in proliferation of MCF-7 breast cancer cells, as did the antisense ER. The antiproliferative effects of the dominant negative and antisense ERs are explained by an increase in the number of cells in the G0/G1 stage of the cell cycle and decrease in the number of cells in G2/M as determined by flow cytometry, and also by a significant increase in the percentage of cells undergoing apoptosis. Our data strongly support the idea that targeting ER action using recombinant viral delivery of dominant negative ERs is an effective way to suppress ER-positive breast cancer cell proliferation and suggests the potential attractiveness of dominant negative gene therapy approaches targeted to the ER for the treatment of hormone-responsive breast cancer. PMID- 10379897 TI - Potential regulation of membrane trafficking by estrogen receptor alpha via induction of rab11 in uterine glands during implantation. AB - The steroid hormone estrogen profoundly influences the early events in the uterus leading to embryo implantation. It is thought that estrogen triggers the expression of a unique set of genes in the preimplantation endometrium that in turn control implantation. To identify these estrogen-induced genes, we used a delayed implantation model system in which embryo attachment to endometrium is dependent on estrogen administration. Using a mRNA differential display (DD) method, we isolated a number of cDNAs representing mRNAs whose expression is either turned on or turned off in response to an implantation-inducing dose of estrogen. We identified one of these cDNAs as that encoding rab11, a p21ras-like GTP-binding protein (G protein), which functions in the targeting of transport vesicles to the plasma membrane. In normal pregnant rats, rab11 mRNA was expressed at low levels on days 1-2 of pregnancy, but its expression was markedly enhanced (approximately 6- to 8-fold) between days 3-5 immediately before implantation. In situ hybridization and immunocytochemistry revealed that rab11 expression in the uterus was predominantly in the glandular epithelium. In ovariectomized rats, the expression of rab11 mRNA was induced in the endometrium in response to estrogen. To determine whether this effect of estrogen was mediated through its nuclear receptors, we examined rab11 expression in a transformed endometrial cell line, Ishikawa. In transient transfection experiments, we observed that overexpression of estrogen receptor (ER) alpha or beta induced endogenous rab11 mRNA in a hormone-dependent manner. ER bound to an antagonist, ICI 182,780, failed to activate this gene expression. These findings, together with the observation that ER alpha but not ER beta is detected in the glands of the preimplantation uterus, indicate that rab11 is one of the proteins that are specifically induced by estrogen-complexed ER alpha in rat endometrium at the onset of implantation. Our results imply that estrogen, which induces the synthesis of many growth factors and their receptors and other secretory proteins that are thought to be critical for implantation, may also facilitate their transport to the membrane and/or secretion by stimulating the expression of rab11, a component of the membrane-trafficking pathway. This study therefore provides novel insights into the diverse cellular mechanisms by which estrogen, acting via its nuclear receptors, may influence blastocyst implantation. PMID- 10379898 TI - Hoxa-10 regulates uterine stromal cell responsiveness to progesterone during implantation and decidualization in the mouse. AB - Hoxa-10 is an AbdominalB-like homeobox gene that is expressed in the developing genitourinary tract during embryogenesis and in the adult uterus during early pregnancy. Null mutation of Hoxa-10 in the mouse causes both male and female infertility. Defective implantation and decidualization resulting from the loss of maternal Hoxa-10 function in uterine stromal cells is the cause of female infertility. However, the mechanisms by which Hoxa-10 regulates these uterine events are unknown. We have identified two potential mechanisms for these uterine defects in Hoxa-10(-/-) mice. First, two PGE2 receptor subtypes, EP3 and EP4, are aberrantly expressed in the uterine stroma in Hoxa-10(-/-) mice, while expression of several other genes in the stroma (TIMP-2, MMP-2, ER, and PR) and epithelium (LIF, HB-EGF, Ar, and COX-1) are unaffected before implantation. Further, EP3 and EP4 are inappropriately regulated by progesterone (P4) in the absence of Hoxa-10, while PR, Hoxa-11 and c-myc, three other P4-responsive genes respond normally. These results suggest that Hoxa-10 specifically mediates P4 regulation of EP3 and EP4 in the uterine stroma. Second, since Hox genes are implicated in local cell proliferation, we also examined steroid-responsive uterine cell proliferation in Hoxa-10(-/-) mice. Stromal cell proliferation in mutant mice in response to P4 and 17beta-estradiol (E2 was significantly reduced, while epithelial cell proliferation was normal in response to E2. These results suggest that stromal cell responsiveness to P4 with respect to cell proliferation is impaired in Hoxa 10(-/-) mice, and that Hoxa-10 is involved in mediating stromal cell proliferation. Collectively, these results suggest that Hoxa-10 mutation causes specific stromal cell defects that can lead to implantation and decidualization defects apparently without perturbing epithelial cell functions. PMID- 10379899 TI - Molecular characterization of the follicle defects in the growth differentiation factor 9-deficient ovary. AB - Growth differentiation factor-9 (GDF-9), a secreted member of the transforming growth factor-beta superfamily, is expressed at high levels in the mammalian oocyte beginning at the type 3a primary follicle stage. We have previously demonstrated that GDF-9-deficient female mice are infertile because of an early block in folliculogenesis at the type 3b primary follicle stage. To address the molecular defects that result from the absence of GDF-9, we have analyzed the expression of several important ovarian marker genes. The major findings of our studies are as follows: 1) There are no detectable signals around GDF-9-deficient follicles for several theca cell layer markers [i.e. 17alpha-hydroxylase, LH receptor (LHR), and c-kit, the receptor for kit ligand]. This demonstrates that in the absence of GDF-9, the follicles are incompetent to emit a signal that recruits theca cell precursors to surround the follicle; 2) The primary follicles of GDF-9-deficient mice demonstrate an up-regulation of kit ligand and inhibin alpha. This suggests that these two important secreted growth factors, expressed in the granulosa cells, may be directly regulated in a paracrine fashion by GDF 9. Up-regulation of kit ligand, via signaling through c-kit on the oocyte, may be directly involved in the increased size of GDF-9-deficient oocytes and the eventual demise of the oocyte; 3) After loss of the oocyte, the cells of the GDF 9-deficient follicles remain in a steroidogenic cluster that histologically resembles small corpora lutea. However, at the molecular level, these cells are positive for both luteal markers (e.g. LHR and P-450 side chain cleavage) and nonluteal markers (e.g. inhibin alpha and P-450 aromatase). This demonstrates that initially the presence of the oocyte prevents the expression of luteinized markers, but that the absence of GDF-9 at an early timepoint alters the differentiation program of the granulosa cells; and 4) As demonstrated by staining with either proliferating cell nuclear antigen (PCNA) or Ki-67 and TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling) labeling, the granulosa cells of GDF-9-deficient type 3b primary follicles fail to proliferate but also fail to undergo cell death. This suggests that granulosa cells of type 3b follicles require GDF-9 for continued growth and also to become competent to undergo apoptosis, possibly through a differentiation event Thus, these studies have enlightened us as to the paracrine roles of GDF-9 as well as the normal steps of granulosa cell and theca cell growth and differentiation within ovarian follicles. PMID- 10379901 TI - Iron-induced oxidative stress in erythrocyte membranes of non-insulin-dependent diabetic Nigerians. AB - The presence of higher level of endogenous free radical reaction products in the erythrocyte ghost membrane (EGM) of Non-insulin-dependent diabetes mellitus (NIDDM) subjects compared with that of normal healthy controls has been demonstrated. The EGMs of NIDDM subjects were also shown to be more susceptible to exogenously generated oxidative stress than those of normal healthy individuals. The decreased level of reactive thiol groups in the EGM of NIDDM individuals supported this observation. We propose that the presence of significant levels of non-heme iron in the EGM of NIDDM subjects is an indication of the potential for iron-catalysed production of hydroxy and other toxic radicals which could cause continuous oxidative stress and tissue damage. Oxygen free radicals could therefore be responsible for most of the erythrocyte abnormalities associated with non-insulin-dependent diabetes and could indeed be intimately involved in the mechanism of tissue damage in diabetic complications. PMID- 10379902 TI - Reversal of sodium arsenite inhibition of rat liver microsomal Ca2+ pumping ATPase by vitamin C. AB - Sodium arsenite (NaAsO2), at 10% of its median lethal dose, was administered to rats with and without vitamin C pretreatment. Liver microsomal fraction was isolated and the activity of Ca2+-ATPase was assayed. Sodium arsenite was found to inhibit the activity of the liver microsomal Ca2+-ATPase to 50% to that of control rats. The specific activity of the enzyme in rats administered sodium arsenite with vitamin C pretreatment was not significantly different from that of control rats. PMID- 10379900 TI - Paracrine actions of growth differentiation factor-9 in the mammalian ovary. AB - Although the transforming growth factor-beta (TGF-beta) superfamily is the largest family of secreted growth factors, surprisingly few downstream target genes in their signaling pathways have been identified. Likewise, the identities of oocyte-derived secreted factors, which regulate important oocyte-somatic cell interactions, remain largely unknown. For example, oocytes are known to secrete paracrine growth factor(s) which are necessary for cumulus expansion, induction of hyaluronic acid synthesis, and suppression of LH receptor (LHR) mRNA synthesis. Our previous studies demonstrated that absence of the TGF-beta family member, growth differentiation factor-9 (GDF-9), blocks ovarian folliculogenesis at the primary follicle stage leading to infertility. In the present study, we demonstrate that mouse GDF-9 protein is expressed in all oocytes beginning at the type 3a follicle stage including antral follicles. To explore the biological functions of GDF-9 in the later stages of folliculogenesis and cumulus expansion, we produced mature, glycosylated, recombinant mouse GDF-9 using a Chinese hamster ovary cell expression system. A granulosa cell culture system was established to determine the role of GDF-9 in the regulation of several key ovarian gene products using semiquantitative RT-PCR. We find that recombinant GDF-9 induces hyaluronan synthase 2 (HAS2), cyclooxygenase 2 (COX-2), and steroidogenic acute regulator protein (StAR) mRNA synthesis but suppresses urokinase plasminogen activator (uPA) and LHR mRNA synthesis. Consistent with the induction of StAR mRNA by GDF-9, recombinant GDF-9 increases granulosa cell progesterone synthesis in the absence of FSH. Since induction of HAS2 and suppression of the protease uPA in cumulus cells are key events in the production of the hyaluronic acid-rich extracellular matrix which is produced during cumulus expansion, we determined whether GDF-9 could mimic this process. Using oocytectomized cumulus cell-oocyte complexes, we show that recombinant GDF-9 induces cumulus expansion in vitro. These studies demonstrate that GDF-9 can bind to receptors on granulosa cells to regulate the expression of a number of gene products. Thus, in addition to playing a critical function as a growth and differentiation factor during early folliculogenesis, GDF-9 functions as an oocyte-secreted paracrine factor to regulate several key granulosa cell enzymes involved in cumulus expansion and maintenance of an optimal oocyte microenvironment, processes which are essential for normal ovulation, fertilization, and female reproduction. PMID- 10379904 TI - Tetramethyl rhodamine methyl ester (TMRM) is suitable for cytofluorometric measurements of mitochondrial membrane potential in cells treated with digitonin. AB - A new method for cytofluorometric analysis of mitochondrial membrane potential deltapsi has been developed by using TMRM as a cationic, mitochondrial selective probe. The method is based on limited treatment of cultured cells with digitonin which permeabilises the plasma membrane and leaves mitochondria intact. The resulting signal of TMRM-stained cells thus represents only the probe accumulated in mitochondria. Fibroblasts and cybrids were used as a model cell systems and optimal conditions for digitonin treatment and staining by TMRM were described. The TMRM signal collapsed by valinomycin, KCN and antimycin A and FCCP titration was used to gradually lower deltapsi and characterise the stability of deltapsi. The method is suitable for sensitive measurement of deltapsi in different types of cultured cells. PMID- 10379903 TI - Effect of isoproterenol on the L-type Ca2+ current in cardiac cells from rats and hybernating ground squirrels. AB - The perforated patch clamp method was used to study the effect of the agonist of beta-adrenoreceptors isoproterenol on L-type Ca2+ current in cardiocytes of rats and ground squirrels in two states: active and hibernating. It is shown that isoproterenol exerts a dual effect on Ca2+ currents of rats and ground squirrels in the active state: at Vh = -50 mV, the current increases, whereas at Vh = -30 mV, it decreases. In hibernating ground squirrels, the dual effect of isoproterenol is not observed: isoproterenol increases Ca2+ current at any Vh values. The hypothesis is put forward that, during the entrance of ground squirrels into hibernation, the phosphorylation of one of the sites (not cAMP dependent) of L-type Ca2+ channels is blocked. PMID- 10379905 TI - Interleukin-6 induced suppression of bovine parathyroid hormone secretion. AB - Calcium disturbances in the critically ill coincide with elevations of proinflammatory cytokines. The effects of tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) on parathyroid hormone (PTH) secretion were investigated. IL-6 and TNF-alpha had no acute effect on PTH secretion in extracellular Ca2+ concentrations of 0.5, 1.25 and 3.0 mM. In contrast to TNF alpha, cultures for 24 h in the presence of 10 ng/mL of IL-6 showed decreased PTH secretion by 51% and 29% in 0.5 mM and 1.25 mM Ca2+ respectively. Neither IL-6 nor TNF-alpha, affected cytoplasmic Ca2+ of the cells. We conclude that PTH secretion in vitro can be suppressed by IL-6 at clinically relevant concentrations. This suppression may aggravate hypocalcemia of the critically ill and attenuate the conventionally strong stimulation of the PTH release by reduction in serum calcium. PMID- 10379906 TI - Uptake of the herbicidal glyphosate by Escherichia coli K-12. AB - The uptake of the aminoacid biosynthesis inhibitor, used as the broad-spectrum herbicide ingredient, glyphosate (N-[phosphonomethyl]-glycine) was investigated in E. coli as a model to study mechanisms of cell resistance to antimetabolites as drugs and pesticides. Unlike the glyphosate-degrading Arthrobacter sp. strain for which the first successful measurement of glyphosate uptake and its inhibition by orthophosphate was reported, E. coli K-12 cannot take up this inhibitor either in the presence of orthophosphate, or after a prolonged starvation for it. However, cells made "competent" after an overnight cold CaCl2 exposure followed by dimethyl sulfoxide (DMSO) treatment could take up this compound (Km for glyphosate uptake, 274 microM). Neither amino acids, belonging to a single transport system, nor orthophosphate gave essential inhibition of glyphosate uptake by these cells. PMID- 10379907 TI - Virus-encoded modulators of cytokines and growth factors. PMID- 10379908 TI - Insulin-like growth factor-1 (IGF-1) and growth hormone (GH) in immunity and inflammation. AB - In recent years many efforts have been undertaken to elucidate the complex interactions between mediators of the endocrine system and the immune system. The main effector of growth hormone (GH) is insulin-like growth factor-1 (IGF-1), an endocrine mediator of growth and development under physiological conditions. Besides this important function, IGF-1 also plays a prominent role in the regulation of immunity and inflammation. This article will address the involvement of IGF-1 in innate as well as acquired immunity and host-defense. We also discuss the role of IGF-1 in the course of inflammatory disorders, including sepsis and sepsis-induced catabolism as well as degenerative arthritis. Based on recent insights, we finally examine the pathophysiological background, potential pitfalls and perspectives of IGF-1 suppletion therapy in these conditions. PMID- 10379909 TI - TNF receptor associated factors in cytokine signaling. AB - Just four years ago the first two members of a new family of molecules involved in signal transduction by members of the TNF receptor superfamily were described and designated TNF Receptor Associated Factors (TRAFs). In the meantime six human and murine TRAFs as well as a TRAF protein from C. elegans have been molecularly cloned. From our current point of view, TRAF proteins appear to represent multifunctional signal adaptors, tightly embedded in a network of signals culminating in the activation of kinase cascades that finally lead to the activation of c-Jun N-terminal kinase. p38 mitogen activated protein kinase, and the transcription factor NF-kappaB, thereby also affecting the balance between survival and cell death. Some of the activities of the individual TRAF family members may be redundant although transgenic knockout animal models have already shown that crucial signaling pathways for single TRAF molecules in vivo can be defined. PMID- 10379910 TI - Cytokine regulation of cellular adhesion molecule expression in inflammation. AB - Cellular adhesion molecules (CAMs) play an essential role in tethering circulating leukocytes to the vascular endothelium at sites of inflammation. They are also instrumental in enabling leukocytes to transmigrate from blood vessels into adjacent inflamed tissues. In the absence of signals to stimulate expression of CAMs, the adhesive forces between leukocytes and the vascular endothelium are below the threshold level required to tether leukocytes. Research in the last decade has shown that several cytokines, including tumour necrosis factor alpha (TNF alpha) and interleukin-1 beta (IL-1beta), potently increase the expression of many CAMs and thus increase the adhesiveness between leukocytes and the endothelium. The CAM-inducing activity of these cytokines is therefore crucial to the regulation of inflammatory processes. Overactivation of CAM expression is linked to a number of acute and chronic inflammatory conditions, and has led to the rationale of antagonising cytokine activity and or CAM expression in order to treat these conditions. The potential application of 'adhesion' antagonists for the therapy of acute chronic inflammatory conditions is briefly discussed. PMID- 10379911 TI - Interleukin-7: physiological roles and mechanisms of action. AB - Interleukin-7 (IL-7), a product of stromal cells, provides critical signals to lymphoid cells at early stages in their development. Two types of cellular responses to IL-7 have been identified in lymphoid progenitors: (1) a trophic effect and (2) an effect supporting V(D)J recombination. The IL-7 receptor is comprised of two chains, IL-7R alpha and gamma(c). Following receptor crosslinking, rapid activation of several classes of kinases occurs, including members of the Janus and Src families and PI3-kinase. A number of transcription factors are subsequently activated including STATs, c-myc, NFAT and AP-1. However, it remains to be determined which, if any, previously identified pathway leads to the trophic or V(D)J endpoints. The trophic response to IL-7 involves protecting lymphoid progenitors from a death process that resembles apoptosis. This protection is partly mediated by IL-7 induction of Bcl-2, however other IL-7 induced events are probably also involved in the trophic response. The V(D)J response to IL-7 is partly mediated through increased production of Rag proteins (which cleave the target locus) and partly by increasing the accessibility of a target locus to cleavage through chromatin remodeling. PMID- 10379912 TI - The MCP/eotaxin subfamily of CC chemokines. AB - Migration of leukocytes from the bone marrow to the circulation, the primary lymphoid organs and inflammatory sites is directed by chemokines and specific receptor interactions. Besides the role of this group of low molecular weight cytokines in leukocyte attraction and activation, anti-HIV and hematopoietic activities were also attributed to chemokines. On the basis of the number and arrangement of the conserved cysteines, chemokines are subdivided in two multi member families, namely the CXC and CC chemokines, whereas fractalkine (CX3C) and lymphotactin (C) are unique relatives. The CC chemokines possess four cysteines of which the first two are adjacent. Functionally, they form a rather heterogeneous family. Here, the focus is on the monocyte chemotactic proteins and eotaxin which, on a structural basis, can be considered as a CC chemokine subfamily. Not only the protein sequences, but also the gene structures, chromosomal location, biological activities and receptor usage exhibit considerable similarities. The review is complemented with a comparison of the biological functions of the MCP/eotaxin-subfamily in physiology and pathology. PMID- 10379913 TI - Analysis of cell death in the trochlear nucleus of the chick embryo: calibration of the optical disector counting method reveals systematic bias. AB - Detection of changes in numbers of neurons is essential for an understanding of neuronal development, function, and death. Optical disector counting is claimed to be the most efficient technique to estimate accurate numbers of neurons in microscopic sections. We calibrated the optical disector by comparison with three dimensional reconstructions from serial sections and determined how accurate this technique is relative to conventional profile counting methods. The calibration was performed on the trochlear nucleus in developing chicks. Optical disector estimates, when obtained as generally recommended, were about 25% lower than the actual number of neurons. This underestimate was caused by a nonuniform (bimodal) distribution of neuronal nuclei in paraffin and plastic (glycolmethacrylate) sections, but not in cryosections. The density of neurons in the core of the paraffin and plastic sections was substantially lower than in the upper and lower margins of these sections. Accurate estimates of neuronal numbers were obtained with a modified optical disector method that sampled the entire extent of tissue sections. Previous estimates of numbers of trochlear neurons in the developing chick have been controversial. The modified (calibrated) optical disector method revealed that the number of trochlear neurons decreased from about 1,600 at day 8.5 of incubation (embryonic day, [E] 8.5) to about 900 at the time of hatching. Numbers of pyknotic nuclei peaked at E6 and at E9, revealing an additional early, but postproliferative, period of cell death. Taken together, these data emphasize the need for calibration of stereological counting techniques and the need to examine sampling strategies for potential bias. PMID- 10379914 TI - Cellular and subcellular distribution of the serotonin 5-HT2A receptor in the central nervous system of adult rat. AB - Light and electron microscope immunocytochemistry with a monoclonal antibody against the N-terminal domain of the human protein was used to determine the cellular and subcellular localization of serotonin 5-HT2A receptors in the central nervous system of adult rat. Following immunoperoxidase or silver intensified immunogold labeling, neuronal, somatodendritic, and/or axonal immunoreactivity was detected in numerous brain regions, including all those in which ligand binding sites and 5-HT2A mRNA had previously been reported. The distribution of 5-HT2A-immunolabeled soma/dendrites was characterized in cerebral cortex, olfactory system, septum, hippocampal formation, basal ganglia, amygdala, diencephalon, cerebellum, brainstem, and spinal cord. Labeled axons were visible in every myelinated tract known to arise from immunoreactive cell body groups. In immunopositive soma/dendrites as well as axons, the 5-HT2A receptor appeared mainly cytoplasmic rather than membrane bound. Even though the dendritic labeling was generally stronger than the somatic, it did not extend to dendritic spines in such regions as the cerebral and piriform cortex, the neostriatum, or the molecular layer of the cerebellum. Similarly, there were no labeled axon terminals in numerous regions known to be strongly innervated by the immunoreactive somata and their axons (e.g., molecular layer of piriform cortex). It was concluded that the 5-HT2A receptor is mostly intracellular and transported in dendrites and axons, but does not reach into dendritic spines or axon terminals. Because it has previously been shown that this serotonin receptor is transported retrogradely as well as anterogradely, activates intracellular transduction pathways and intervenes in the regulation of the expression of many genes, it is suggested that one of its main functions is to participate in retrograde signaling systems activated by serotonin. PMID- 10379915 TI - Descending auditory pathways: projections from the inferior colliculus contact superior olivary cells that project bilaterally to the cochlear nuclei. AB - Multiple retrograde and anterograde tracers were used to characterize a pathway that extends from the inferior colliculus to both the left and right cochlear nuclei via a synaptic relay in the superior olivary complex. Different fluorescent tracers were injected into the left and right cochlear nuclei to identify cells in the superior olivary complex that project bilaterally. Double labeled cells were present in almost all periolivary nuclei; the majority were located in the ventral nucleus of the trapezoid body and the anteroventral periolivary nucleus. Because these two nuclei are targets of descending projections from the inferior colliculus, triple-labeling experiments were performed to determine whether collicular axons contact the periolivary cells that project to the cochlear nuclei. The results demonstrate that descending axons from the inferior colliculus contact periolivary cells that project to the cochlear nuclei, including periolivary cells that project bilaterally. This pathway could provide an opportunity for higher levels of the auditory system to influence activity bilaterally in the cochlear nuclei and thus to modulate the initial processing of acoustic information by the brain. PMID- 10379916 TI - Seasonal changes in androgen receptor immunoreactivity in the song nucleus HVc of a wild bird. AB - In seasonally breeding songbirds, song behavior and neural morphology change seasonally. Song control nuclei are larger during the breeding season, as determined by multiple cytological labels. Seasonal changes in song nuclei are regulated by testosterone (T), and several song nuclei contain intracellular androgen receptors (AR). Changes in AR levels may interact with changes in plasma T levels to regulate song nuclei morphology. We measured seasonal changes in AR immunoreactive cells in the telencephalic song nucleus HVc using the affinity purified PG21 antibody to rat AR. We caught wild adult male Gambel's white crowned sparrows (Zonotrichia leucophrys gambelii) during spring breeding in Alaska and during autumn migration in Washington State. To enhance PG21 labeling, animals were treated with T for 90 minutes (as in Smith et al. [1996] J. Histochem. Cytochem. 44:1075-1080). AR+ cells were found in HVc and other song nuclei, hippocampus, nucleus taeniae (homologue to mammalian amygdala), and the hypothalamus. HVc volume was larger in spring (S) than autumn (A), in both the PG21- and Nissl-stained sections (S:A = 1.9 and 1.7, respectively). In spring, but not autumn, PG21 and Nissl measurements were slightly different (PG21:Nissl = 1.07), perhaps because PG21 labeled the most caudal extent of HVc more clearly. In HVc, AR+ cell density and number were greater in spring. The percentage of AR+ cells was also increased in spring. Qualitatively, the staining intensity of individual cells was higher in spring. In time course studies, the T injection enhanced PG21 staining within 15 minutes, suggesting that it increases labeling via AR translocation to and concentration in the cell nucleus. PMID- 10379917 TI - NMDA receptor NR1 and NR2A/B subunit expression in trigeminal neurons during early postnatal development. AB - Trigeminal motoneurons (Mo5), mesencephalic trigeminal neurons (Me5), and supratrigeminal (Su5) and intertrigeminal (15) neurons are important constituents of the neural circuitry responsible for jaw movements observed during ingestive behaviors. In addition, in adult animals, N-methyl-D-aspartate (NMDA) receptors are a critical component of the brainstem circuitry responsible for reflex- and centrally activated jaw movements. However, little is known about the expression of this receptor in circuitry used to produce neonatal jaw movements. Receptor immunohistochemistry was used to describe changes in the expression of NMDA NR1 and NR2A/B receptor subunits in Mo5, Me5, Su5, and I5 neurons during postnatal development. Rats at postnatal days (P) 1, 3, 8, 15-16, 21-24, and 28-35 were used. An affinity-purified polyclonal antibody against the NR1 subunit and an affinity-purified polyclonal antibody that recognizes both NR2A and 2B subunits were used to depict the expression of these subunits. In Mo5, immunoreactivity was noted for both antibodies throughout the time frame sampled. NR1 expression in Me5 neurons emerged at P1. NR2A/B expression emerged at P3 in caudal and middle regions of Me5 and at P8 for rostral regions of the nucleus. NR1 immunoreactivity was present at P1 for neurons in I5 and at P3 for neurons in the Su5 region. NR2A/B subunit expression in Su5 and 15 neurons emerged at P8. These results provide evidence for NMDA receptor subunits in neonatal trigeminal neurons used in oral-motor circuitry and suggest a role for the NMDA receptor in synaptogenesis associated with these neurons during postnatal development. PMID- 10379918 TI - Dendritic localization of Ca(2+)-permeable AMPA/kainate channels in hippocampal pyramidal neurons. AB - Although it is well established that cortical and hippocampal gamma-aminobutyric acid (GABA)-ergic neurons generally have large numbers of Ca(2+)-permeable alpha amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)/kainate channels (Ca A/K channels), their presence on pyramidal neurons is controversial. Ca2+ permeability of AMPA channels is regulated by expression of a particular glutamate receptor subunit (GluR2), which confers Ca2+ impermeability to heteromeric channels. Most electrophysiology studies, as well as in situ hybridization and immunolabeling studies demonstrating expression of GluR2 mRNA or peptide in pyramidal neurons, have provided evidence against the presence of Ca-A/K channels on pyramidal neurons. However, observations that pyramidal neurons often appear to be labeled by kainate-stimulated Co2+ influx (Co2+(+) cells), a histochemical stain that identifies cells possessing Ca-A/K channels, suggests that they may have these channels. The present study futher examines cellular and subcellular distribution of Ca-A/K channels on hippocampal pyramidal neurons in slice as well as in culture. To this end, techniques of kainate stimulated Co2+ influx labeling, supplemented by AMPA receptor subunit immunocytochemistry and fluorescent imaging of kainate-stimulated intracellular Ca2+ ([Ca2+]i) rises are employed. Co2+ labeling is often seen in pyramidal neuronal dendrites in both slice and in culture. In addition, although GluR1 and 4 staining in these neurons is often seen in the soma and dendrites, GluR2 label, when evident, is generally more restricted to the soma. Finally, measurement of kainate-stimulated [Ca2+]i rises in cultured neurons, assessed by using low affinity Ca2+ indicators in the presence of N-methyl-D-aspartate (NMDA) receptor and voltage-sensitive Ca2+ channel blockade, often shows dendritic rises to precede those in the somata. Thus, these data support the hypothesis that Ca-A/K channels are present in dendritic domains of many pyramidal neurons, and may help to provide resolution of the apparently conflicting data regarding their distribution. PMID- 10379919 TI - Development of inhibitory circuitry in visual and auditory cortex of postnatal ferrets: immunocytochemical localization of GABAergic neurons. AB - The goal of this study was to describe the development of gamma-aminobutyric acid (GABA)-containing neurons in visual and auditory cortex of ferrets. The laminar and tangential distribution of neurons containing excitatory, inhibitory, and neuromodulatory substances constrain the potential circuits which can form during development. Ferrets are born at an early stage of brain development, allowing examination of inhibitory circuit formation in cerebral cortex prior to thalamocortical ingrowth and cortical plate differentiation. Immunocytochemically labelled nonpyramidal GABA neurons were present from postnatal day 1 throughout development, in all cortical layers, and generally followed the inside-out pattern of neuronal migration into the cortical plate. Prior to postnatal day 14, pyramidal neurons with transient GABA immunoreactivity were also observed. The density of Nissl-stained and GABA-immunoreactive neurons was high early in development, declined markedly by postnatal day 20, then remained relatively constant until adulthood. However, examination of the proportion of GABA neurons revealed an unexpected late peak at postnatal day 60, then a decrease in adulthood. Visual and auditory cortex were similar in most respects, but the peak at postnatal day 60 and the final proportion of GABA neurons was higher in auditory cortex. The late peak suggests that inhibitory circuitry is stabilized relatively late in sensory cortical development, and thus that GABA neurons could provide an important substrate for experience-dependent plasticity at late stages of development. PMID- 10379920 TI - Nitric oxide-containing neurons in the nervous ganglia of Helix aspersa during rest and activity: immunocytochemical and enzyme histochemical detection. AB - Nitric oxide synthase (NOS) immunoreactivity and staining for nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-diaphorase) activity are two cytochemical markers for nitric oxide (NO)-containing neurons. The authors examined the changes in the distribution of NOS immunolabeling and NADPH diaphorase reactivity in the cerebral and buccal ganglia of the terrestrial snail Helix aspersa during resting and active phases. During inactivity and after 1 day of activity, in the mesocerebrum and metacerebrum of the snails, there were several reactive neurons for both markers; after 7 days of activity, the number of reactive neurons was lower. Opposite results were obtained in the buccal ganglia, in which increased staining and numbers of reactive neurons were present in the active snails (after 1 day and 7 days of activity). Although the staining patterns for the two reactions were similar, colocalization was not always observed. The comparison between inactive and active animals provided a more precise survey of NOS-containing neurons in the snail cerebral ganglia than previously described. Moreover, it suggested that not only is NO involved in distinct nervous circuits, but, as a ubiquitous molecule, it also plays a role in neuroprotection and neuropeptide release. PMID- 10379921 TI - Ventrally located commissural neurons express the GABAergic phenotype in developing rat spinal cord. AB - Early-forming commissural neurons are studied intensively as a model of axonal outgrowth and pathfinding, yet the neurotransmitter phenotype of the majority of these neurons is not known. The present study has determined that a substantial number of commissural neurons express the 65-kDa isoform of glutamic acid decarboxylase (GAD65) as early as embryonic day 12 (E 12). Patterns of GAD65 localization were compared with those of TAG-1, the Transiently expressed Axonal Glycoprotein that is the best known marker of commissural axons. On E13, both GAD65- and TAG-1-labeled commissural axons emanate from similar lateral and ventromedial regions. However, dorsally located TAG-1-positive commissural axons were GAD65-negative. These results suggest that commissural neurons have both gamma-aminobutyric acid (GABA)ergic and non-GABAergic phenotypes. The intensity of GAD65 staining within commissural somata and axons decreased between E14-15 and continued to decline during embryonic development, whereas terminal-like structures in surrounding neuropil increased dramatically. This sudden loss of somatic and axonal GAD65 staining was unexpected and could be interpreted as commissural neurons only transiently expressing the GABAergic phenotype. Further experiments were undertaken to identify commissural neurons with other established GABAergic markers, GAD67 and GABA. When antibody labeling of the two GAD isoforms was compared, GAD67 was detected 1 day later than GAD65, and in a different subcellular distribution. In contrast to GAD65, GAD67 intensely stained somata but labeled few commissural axons. GABA immunoreactivity also was detected in commissural axons 1 day after GAD65, and the labeling pattern between E13 and E16 resembled that of GAD67 rather than GAD65. When GAD and GABA results were compared, it was clear that a number of ventrally located commissural neurons expressed and maintained the GABAergic phenotype during embryonic development. However, the early expression and subcellular redistribution of GAD65 suggests that the GAD isoforms are differentially regulated. The function of the transient GAD65 expression in commissural somata and axons is unknown, but its temporal expression pattern parallels the transient expression of TAG-1, as both are expressed during the early stages of commissural axon outgrowth and pathfinding. PMID- 10379922 TI - Large-scale synaptic errors during map formation by regeneration optic axons in the goldfish. AB - During the formation of visual maps, growing axons initially form a map by using topographically distributed cues that direct their growth and branching to the appropriate target region. This initial map is typically roughly retinotopic and is subsequently refined through activity-dependent rearrangement or cell death. Although synaptic connections are thought to be rearranged during the later refinement phase, there is no clear evidence that synapses are being formed during the initial targeting phase of development. Also, because optic fiber growth can be accurately directed during normal development, it is unclear whether regenerative fibers that have more pathway disorder would behave similarly. This issue was addressed by using optic fibers of goldfish that have the capacity to regenerate a retinotopic projection and can reestablish a rough retinotopic order without impulse activity. The optic nerve of goldfish was crushed, and at various times later, a small number of optic fibers in ventronasal retina was labeled with wheatgerm agglutinin-horseradish peroxidase. The tectum was then processed for electron microscopy to look at the distribution of labeled synapses during regeneration. At 3 weeks, synapses were observed at the far anterior end of the tectum and none were yet seen at the correct posterior retinotopic position. At 4-5 weeks, synapses were seen in nearly equal numbers at the incorrect anterior end and at both correct (retinotopic) and incorrect posterior positions. At late stages of regeneration, synapses were restricted to their correct posterior retinotopic position in the tectum, as they were in normal fish. These findings show that the formation of global retinotopic order entails the formation and subsequent elimination of a large number of highly ectopic synapses. Synaptic rearrangement is a major feature of targeting in this system and may be required for the regeneration of a retinotopic projection. PMID- 10379924 TI - Thoracic and prothoracic leg neuromuscular system of the praying mantid, Sphodromantis lineola (Burmeister). AB - Historically, praying mantids have attracted attention because of their dramatic prey capture behavior, loosely termed the strike. However, little is known about the neuromuscular organization that underpins the behavior. Although once thought to be quite stereotyped, recent data indicate that the strike is quite plastic and can be aimed accurately within a relatively large three-dimensional space. Hence, successful prey capture requires the integration of (1) visual information, indicating prey has been recognized; and (2) proprioceptive information, indicating head and prothorax (i.e., visual field) position and initial leg positions. This study was undertaken as part of a larger program examining how such sensory information is integrated with the appropriate motor systems. Our goals were (1) to describe the gross thoracic and foreleg neuromuscular system of Sphodromantis lineola and (2) to identify the soma locations of the motor neurons associated with the largest leg nerve, N4, which travels the length of each leg. We found that the thoracic and foreleg neuromusculature of S. lineola are similar but not identical to what is known about just three other species of mantid, and that motor neuron somata associated with N4 are arranged in stereotypical, bilaterally symmetrical groups as they are in other orthopteroids, suggesting that this is a general organizational feature of the insect CNS. PMID- 10379923 TI - Central melatonin receptors in the rainbow trout: comparative distribution of ligand binding and gene expression. AB - To better define the role of melatonin in fish, we have compared in detail the distribution of 2-[125I]iodomelatonin binding sites with gene expression for melatonin receptor subtypes in a widely studied seasonal species, the rainbow trout. Three distinct partial sequences of the melatonin receptor gene were cloned from trout genomic DNA. Two of the sequences corresponded to the Mella receptor subtype, and one corresponded to the Mellb receptor subtype. Analysis of numerous clones failed to find a sequence equivalent to the Mel1c receptor subtype. Comparison of receptor gene expression with 2-[125I]iodomelatonin binding distribution indicated dendritic transport of the receptor. Melatonin receptors were associated predominantly with visually related areas of the trout brain, such as the thalamic region, the pretectal area, and the optic tectum. The pituitary was devoid of 2-[125I]iodomelatonin binding, and melatonin receptor gene expression was not detectable. It would appear from the results of the present study that melatonin in this species is involved primarily in the processing of visual signals. How melatonin interacts with circannual rhythms of growth and reproduction is unclear, although a direct interaction between melatonin and the hypothalamo-pituitary axis is not clearly indicated. PMID- 10379925 TI - Different contractile properties between intralobar and extralobar pulmonary arteries of the rabbit. AB - In pulmonary circulation, small muscular resistance arteries are known to have different receptor properties and sensitivity to neurotransmitters from those of large elastic conduit arteries. It is, however, not yet certain whether the different properties are primarily due to the diameter or the location of arteries. In the present study, we compared the contractile responses to various agonists among large extralobar (ELPA, diameter: 2-3 mm), large intralobar (ILPA, diameter: 2-3 mm), and small intralobar pulmonary arteries (SPA, diameter: 300 500 microm) of the rabbit. There were no differences in normalized dose-response curves to KCl among three groups. Half maximum doses (EC50 in mM) were 38.0+/-2.0 (n=8, mean+/-SEM) in ELPA, 36.9+/-2.4 (n=10) in ILPA, and 39.0+/-0.9 (n=12) in SPA. Responses to phenylephrine, epinephrine, histamine, serotonin, and PGF2alpha were normalized and expressed as a relative contraction against maximum tension to KCl. In ELPA, the contractile responses to various agents showed the following sequence: KCl>epinephrine>phenylephrine>serotonin>PGF2alpha>histamine. In ILPA and SPA, the sequence was: KCl>histamine>PGF2alpha>serotonin. There was little response to phenylephrine and epinephrine in ILPA and SPA. These results demonstrate that the difference of contractile responses between ELPA and ILPA was more prominent than that between ILPA and SPA, suggesting that the location is more important than the diameter itself in determining the characteristic contractile responses of pulmonary arteries. PMID- 10379926 TI - Comparative studies on the relaxing action of several adenosine 5'-triphosphate sensitive K+ channel openers in pig urethra. AB - The relaxing effects of the adenosine 5'-triphosphate (ATP)-sensitive K+ channel openers (K(ATP) openers; diazoxide, minoxidil, pinacidil, (+/-)-cromakalim, (+) cromakalim and (-)-cromakalim) were investigated on the resting tone of pig proximal urethra. In addition, patch clamp techniques were utilized for recording cromakalim-induced ionic currents in cells dispersed from the same urethral region. The (-)-cromakalim-induced relaxation of urethral muscle strips was stable, reversible and reproducible. The rank order of potency regarding of K(ATP) openers in lowering the resting urethral tone was (-) cromakalim>pinacidil>diazoxide>minoxidil. K(ATP) opener-induced urethral relaxation was suppressed by subsequent application of glibenclamide (1 microM). (+)-Cromakalim (< or =10 microM) did not relax the urethra nor antagonize the (-) cromakalim-induced urethral relaxation. However, at higher concentrations, (+) cromakalim (> or =30 microM) caused a small but significant urethral relaxation. In accordance with these observations, the relaxation induced by 5 microM (-) cromakalim was identical to that induced by 10 microM (+/-)-cromakalim, as expected from a theoretical half potency for (+/-)-cromakalim. In whole-cell recording, (-)-cromakalim and (+) cromakalim (100 microM) activated a glibenclamide-sensitive outward current which was due to the activation of the glibenclamide-sensitive 43 pS K+ channel (K(GS)-43 pS). The potency of (+) cromakalim to activate K(GS)-43 pS was much weaker than that of (-)-cromakalim. These results indicate that the ability of K(ATP) openers to relax pig urethral smooth muscle can be accounted for by activation of glibenclamide-sensitive K+ channels. PMID- 10379927 TI - Absence of ganglionated plexus in bullfrog gallbladder. AB - We first carried out microscopic observation of the intramural nerves of a bullfrog gallbladder which were fixed and stained with a solution of OsO4 and ZnI2. We then investigated if the responses of isolated frog gallbladder evoked by electrical stimulation are mediated through the intramural nerves. The following results were obtained: 1. The nerve plexus and the perivascular nerves were observed in the subserosal layer of the wall of the gallbladder. These nerves do not have a ganglia. That is to say, no ganglionated plexus or ganglia were observed in the subserosal layer of the wall of the gallbladder. 2. Electrical stimulation caused the gallbladders to contraction with rectangular pulses (50 volt, 40 Hz) of durations of 0.5, 1, 2, 3, 4 and 5 msec for a period of 10 sec. Three blockers of nerve mediated responses, atropine (1 x 10(-6) M), guanethidine (1 x 10(-6) M) and tetrodotoxin (3 x 10(-7) M), had no effect on the gallbladder contractions induced by stimulation with pulses as short as 0.5 msec or as long as 5 msec. These results suggest that the bullfrog gallbladder may not contain nerves related to movement. Thus, the contraction of the bullfrog gallbladder induced by electrical stimulation does not seem to be modulated by extrinsic nerve terminals distributed in the gallbladder wall. PMID- 10379928 TI - Contractile protein isoforms of single and cultured smooth muscle cells from guinea pig ileum. AB - Single smooth muscle cells isolated from guinea pig ileum using collagenase and papain produce contractile response to muscarinic agents, while the cultured cells do not. Using fluo-3/AM and a confocal laser scanning fluorescence microscope, it was observed that carbachol, a muscarinic agent, caused an increase in the intracellular Ca2+ of both single and cultured cells. SDS-PAGE and Western Blot analyses revealed the expression of myosin heavy chain isoforms of SM1 (204 kDa) and SM2 (200 kDa) in single smooth muscle cells, and non muscle isoform (196 kDa) of myosin heavy chain only in the cultured cells. With respect to actin isoforms, alpha-actin was predominant in single cells and beta-actin was major in the cultured cells. Two types of tropomyosin monomer, 39 kDa and 41 kDa, were detected in single cells, while the 41 kDa monomer was lost in cultured cells. These differences in contractile protein profiles between single and cultured cells were collaborated with the observation of cells using immunofluorescence microscope with responsible antibodies to isoforms of myosin heavy chain, actin and tropomyosin. These results suggest that the loss of contractility in cultured smooth muscle cells is profoundly related to changes in contractile protein profiles from smooth muscle type to non muscle type. PMID- 10379929 TI - Influences of endothelium on the time course of noradrenaline-, 5-HT-, prostaglandin F2alpha- and high-K+-induced contractions in aortae of WKY and SHRSP. AB - Influences of endothelium on contraction of aortic smooth muscle by various agents were studied and those in the preparations from Wistar Kyoto rat (WKY) and stroke-prone spontaneously hypertensive rats (SHRSP) were compared. Endothelium depressed the contractions induced by noradrenaline and 5-hydroxytryptamine (5 HT). The time course of the contraction was bi- or tri-phasic in the former and slow rising monophasic in the latter. On the other hand, the depression was weaker in the contraction by prostaglandin F2alpha- and high K+. The depression was blocked by the removal of endothelium or in the presence of Nomega-nitro-L arginine (L-NNA), indicating that nitric oxide (NO) released from endothelium was responsible for the inhibition. The inhibition was weaker in the preparation from SHRSP when compared to that in the preparation from WKY. Relaxation by acetylcholine (ACh) of the preparation precontracted in the presence of respective contractile agent was impaired in the preparation from SHRSP. It is concluded that mode of inhibition of the contraction varies depending on the agents used to initiate the contraction, i.e. depending on the mode of the release of NO. In the preparation from SHRSP, the influence of endothelium is impaired due to the reduced release of NO. PMID- 10379930 TI - Involvement of endothelium-derived factors in controlling the active tone of smooth muscle in aorta from hypertensive rats. AB - Control of the active tone by endothelium in aortae from various strains of spontaneously hypertensive rats was studied. The active tone was negligibly observed in endothelium-intact preparation. The application of N(G)-nitro-L arginine (L-NNA, 10(-4) M) induced slowly developed active tone in the preparations from hypertensive rats but no active tone was induced in the preparation from normotensive Wistar Kyoto rats (WKY). The developed tension was stronger in preparations from rats with higher blood pressure as observed in endothelium denuded preparations. The developed active tone in the presence of L NNA was greater than that observed in endothelium denuded preparations. The active tone was abolished by the removal of extracellular Ca2+ or by the application of Ca-antagonists. L-arginine counteracted the effects of L-NNA and depressed the developed active tone in the presence of the latter drug. The application of indomethacin (10(-5) M) depressed the active tone of the preparations from SHRSP by 25.5+/-5.2%. Increasing extracellular K+ concentration or application of tetraethylammonium (TEA) could not be used to observe the effect of endothelium-derived factors on the active tone, because of their strong contractile effect. Simultaneous application of apamin and charybdotoxin induced an elevation of tension which was often associated with spontaneous tension oscillation. It is concluded that the active tone, which is smooth muscle origin, is depressed by endothelium-derived nitric oxide (NO) strongly and potentiated by a product of arachidonic acid cascade through cyclooxygenase pathway. The involvement of endothelium-derived hyperpolarizing factor (EDHF) in the depressing effect of endothelium is thought to be small. PMID- 10379931 TI - Beta-adrenoceptors in the detrusor of guinea pig bladder. AB - In the present study, we tried to determine which beta-adrenoceptor subtypes are involved in the inhibitory regulation of bladder motility in the guinea pig by using in vitro functional analysis. Isoprenaline, norepinephrine and epinephrine decreased spontaneous contractile activity of the bladder concentration dependently, the rank order of their potency being isoprenaline (pD2 = 7.38)>norepinephrine (6.71)>epinephrine (6.31). Dobutamine was also effective in inhibiting the spontaneous contraction (5.81). However, CGP-12177, BRL37344, salbutamol, and clenbuterol were not effective at the concentration of 30 microM. The concentration response curves of catecholamines (isoprenaline, norepinephrine and epinephrine) were rightward shifted by the presence of atenolol. Schild regression analyses carried out for atenolol against isoprenaline, norepinephrine and epinephrine gave pA2 values of 6.93, 6.78 and 6.69, respectively. The concentration-response curve for isoprenaline was unaffected by 10 microM butoxamine. In the presence of 1 microM propranolol, bupranolol produced shifts of the concentration-response curve for isoprenaline, and the apparent pA2 value for bupranolol against isoprenaline was 5.50. These results suggest that the inhibition of spontaneous contraction of the guinea pig bladder by beta adrenoceptor agonists is mediated mainly via beta1-adrenoceptor and the effect of isoprenaline is mediated partly via atypical beta-adrenoceptor subtype. PMID- 10379933 TI - Characterization of promoters integrated in the genome of bovine herpesvirus-1 (BHV-1). AB - Bovine herpesvirus-1 (BHV-1) has been used as a vector of live recombinant vaccines for cattle which express the genes of other pathogens. Because of the importance of the choice of the promoter which allows the efficient expression of the foreign genes in the BHV-1 vector, we compared the relative efficacy of various promoters integrated in the BHV-1 genome. The promoter sequences of the BHV-1 thymidine kinase (tk), gB, gC, SV40 early, and pseudorabies virus (PRV) immediate early (IE) genes were placed at the upstream of the open reading frame of the chloramphenycol acetyl transferase (CAT) gene and the promoter-CAT sequences were integrated into the tk gene of BHV-1 by homologous recombination. The promoter activity was assayed by measuring the CAT activity in the extracts of Madin Darby bovine kidney (MDBK) cells infected with the recombinant BHV-1. The PRV IE promoter was activated earlier and maintained at a higher level activity than the BHV-1 gB or gC promoters throughout the most of the growth phase of BHV-1. At the late phase, however, the activities of the BHV-1 gB and gC promoters reached the higher level. The BHV-1 tk promoter activity was low and the SV40 early promoter was hardly activated when integrated into the BHV-1 genome. promoter, recombinant BHV-1. PMID- 10379932 TI - Relations between plasma acetate, 3-hydroxybutyrate, FFA, glucose levels and energy nutrition in lactating dairy cows. AB - To clarify the implication of an energy nutrition on a metabolic alteration with advancing lactation, total 270 blood samples were taken from 16 lactating dairy cows. Amounts of dietary allowance and the refusals were measured daily, and the energy (TDN) intakes and a satisfaction (energy balance) of each cow were estimated. Plasma acetate, 3-hydroxybutyrate (3-HB), free fatty acid (FFA) and glucose levels were estimated. The data were divided into 3 groups depending on the days in milk; early (up to 70 days postpartum), mid (71 to 140 days), and late (after 141 days) lactation. There were many cases of higher FFA level in early lactation, especially with declining acetate and glucose levels. There were proportional elevations of 3-HB in connection with FFA levels in many samples of early lactation, though the 3-HB increased independently of FFA levels in the most cases of the mid and late lactations. Plasma 3-HB levels increased in many cases of decreased glucose level, especially in the early lactation. Plasma acetate level correlated positively with 3-HB level, but not correlated with glucose level. Higher FFA level and elevation of FFA/3-HB ratio were observed in the conditions of negative energy balance. This implies the metabolic importance of FFA in a ketogenesis of the early lactation. PMID- 10379934 TI - Pharmacokinetic and depletion studies of sarafloxacin after oral administration to eel (Anguilla anguilla). AB - The pharmacokinetics of sarafloxacin applied by oral gavage at a dose of 15 mg/kg b.w. was studied in eel (Anguilla anguilla) at water temperature of 24 degrees C. Sarafloxacin levels were determined using high performance liquid chromatography with a quantitation limit of 0.07 microg/ml or gram. The time to peak plasma concentration, Tmax, was 12 hr and peak concentration, Cmax, was 2.64 microg/ml. The absorption rate constant (k(a)) was 0.23 hr(-1) (r=0.996). The drug disposition curve after Tmax was fitted to a two-compartment open model. The distribution rate constant (alpha) was 0.085 hr(-1) (r=0.972), and the half-life (t(1,2alpha)) was 8.15 hr. The elimination rate constant (beta) was 0.023 hr(-1) (r=0.909), and the half-life (t(1/2beta)) was 30.13 hr. The estimated area under the curve, AUC, was 56.7 microg.hr/ml. The peak concentrations of drug in liver, kidney, muscle, and skin were 13.39 (12 hr), 5.53 (12 hr), 1.82 (24 hr), and 0.78 microg/g (40 hr), respectively. The time for sarafloxacin mean levels to fall below detectable limits in the plasma, muscle, and skin were 7 days but for the liver and kidney were 14 days. PMID- 10379935 TI - Changes in lectin binding patterns of mouse male germ cells (gonocytes) during prespermatogenesis. AB - The distribution of sugar residues in gonocytes of the differentiating mouse testis was examined by light microscopy using 22 different kinds of lectins. Characteristic binding patterns of sWGA, VVA, and LEA in gonocytes were observed during prespermatogenesis. sWGA preferentially bound to the cytoplasm and plasma membrane of gonocytes on 16.5 days post coitus (dpc). Its reaction decreased thereafter and almost disappeared on 1.5 days post partum (dpp), but reaction reappeared on 4.5 dpp and continued until 6.5 dpp. The VVA reaction was recognized in a few gonocytes on 0.5 dpp, and remained strong until 6.5 dpp. LEA reacted strongly in the plasma membrane and cytoplasm of gonocytes from 0.5 dpp to 6.5 dpp. The present study indicates that sWGA, VVA, and LEA are useful markers for gonocytes, and the appearance or disappearance of sWGA and VVA may be related to the differentiation of gonocytes during prespermatogenesis. PMID- 10379936 TI - Infectivity to hosts of the endogenous stages of chicken and murine Cryptosporidium. AB - Five groups of 4 mice each were inoculated with 10(6) Cryptosporidium muris oocysts. They were necropsied on days 2, 4, 6, 8 and 10. The stomach mucosa from each group were made into 10% suspension in physiological saline and were orally inoculated to 2 mice each. Recipients given suspension from infected mice on day 6, 8 and 10 shed oocysts from 6, 9 and 6, respectively. Similarly, White Leghorn received 10(6) Cryptosporidium sp. oocysts were killed daily between 1 and 6 days. Recipients given bursa of Fabricius or caecum of donor birds on days 4, 5 and 6 shed oocysts. The endogenous stages of murine and chicken Cryptosporidium were able to infect the appropriate host. PMID- 10379938 TI - Method of lymphocytotoxic crossmatch test for feline renal transplantation. AB - The optimal condition for methods of lymphocytotoxic crossmatch test for feline renal transplantation was investigated. On separation of viable lymphocytes from whole blood, the best results were obtained when Ficoll-diatrizoate with 1.078 of a specific gravity at 20 degrees C was centrifuged with 800 x g for 30 min at 4 degrees C. A nylon wool column was used to separate T and B cells from lymphocyte fraction. The ratio of T cells in nylon wool effluent cells was 95%, while the ratio of B cells in adherent cells was 41%. Lymphocytotoxic crossmatch tests were performed by using the effluent cells as T cells and the adherent cells as B cells, at 37 degrees C (warm) and 4 degrees C (cold). The ratio of B cells in adherent cells was low, however, the result was utilized as a matching test before transplantation by combining with the T cell result. The trypan blue stain method made it easier than the eosin stain method to distinguish living and dead cells. The lymphocytotoxic crossmatch tests were performed on 15 pairs of healthy cats, and only one pair showed doubtful positive against anti-B cell cold antibodies. During acute rejection after renal transplantation in two pairs which were negative on any anti-lymphocyte antibodies before the transplantation, the anti-T cell warm antibodies became positive in both pairs, and the anti-T cell cold antibodies became positive on one of the two pairs. PMID- 10379937 TI - Characterization of immune responses caused by bovine leukemia virus envelope peptides in sheep. AB - To study the immunomodulative activity caused by bovine leukemia virus envelope (BLV Env) peptide, sheep were immunized with two kinds of Th-epitope peptides, peptide 98 (BLV Env 98-117), and 61 (BLV Env 61-78). Four of eight immunized sheep showed specific proliferative responses against both of the peptide stimulations. To characterize the cells responding to the peptides, peptide specific cells were established from the responding sheep by the continuous stimulation of peripheral blood mononuclear cells (PBMCs) with either peptide 98 or 61 in vitro. The peptide 98-specific cells consisted of CD4-positive cells, whereas the peptide 61-specific cells consisted of CD8-positive cells and MHC class II-positive cells. In addition, cytokine profile analysis indicated that the peptide 98-stimulated cells expressed IFN-gamma but not IL-10, although the peptide 61-stimulated cells expressed IL-10 but not IFN-gamma. These results show that BLV envelope peptides 98 and 61 can modulate immune responses of sheep lymphocytes in different ways and may contribute to the pathogenesis of BLV infection. PMID- 10379939 TI - Effects of peripheral blood polymorphonuclear leukocyte function and blood components in Japanese black steers administered ACTH in a cold environment. AB - An examination of the effects of artificial stress induced by adrenocorticotropin (ACTH) on total and differential leukocyte counts, plasma cortisol levels, metabolic profiles and peripheral blood polymorphonuclear leukocyte (PMN) function was performed on Japanese Black steers kept in a cold environment, with the following regimes; 1) -5 degrees C x ACTH (100 IU/day for 3 days), 2) 0 degrees C x ACTH, 3) 15 degrees C x ACTH and 4) 15 degrees C x PBS. Blood samples were collected before and at 1, 2, 24, 48, 72 and 96 hr prior to the application of the stressor. The plasma cortisol level was found to greatly increase at one hr after the first treatment of ACTH, particularly so in animals exposed to -5 degrees C. Total leukocytes (-5 degrees C and 0 degrees C experiments, respectively), the monocytes (-5 degrees C), neutrophils and eosinophils (-5 degrees C, 0 degrees C and 15 degrees C, respectively) obviously increased just after the first administration, although lymphocyte counts at -5 degrees C were inversely related to those described above. All of these tendencies were augmented by the cold environment except for eosinophils. The chemiluminescent (CL) response of PMN decreased in the ACTH-administered steers at an early stage of post-administration, however, it tended to recover from the lower-than-base value in the cold-affected steers. ACTH administration resulted in higher plasma glucose (Glu) compared to a control, although only steers housed at -5 degrees C evidently showed lower plasma inorganic phosphorus (IP). No abnormal serum acute phase protein, or immunosuppressor, was noted. ACTH thus appears not only to promote physiological reactions but also to temporarily suppress PMN cellular immune function in Japanese Black steers. Although, a cold environment rapidly restored the CL activity to over the pre-administrational value, suggesting that a vital response was activated by crymo-stimuli. PMID- 10379940 TI - Intercalated duct cells in the chicken pancreatic islet with special reference to the alloxan administration. AB - The intercalated duct cells were observed in the A and B islets of the chicken pancreas. These cells adhered with each other by intercellular junctional complexes at the apical side. They had many microvilli projecting into the lumen. Abluminally, they displayed extended slender cytoplasmic processes between islet endocrine cells. Administration of alloxan resulted to denser cytoplasm and a more prominent thickening of cytoplasmic processes of the intercalated duct cells, although the blood glucose levels did not show appreciable changes by the treatment. The intercalated duct epithelial cells appeared clearly as stellate cells. The lysosomes increased in size and number with passage of time after alloxan administration. The present findings may suggest that intercalated ducts are not only anatomically important as a structure passing through the islet but also play physiologically by protecting the islet endocrine cells. PMID- 10379941 TI - Molecular cloning of two Haemaphysalis longicornis cathepsin L-like cysteine proteinase genes. AB - Immunological protection of mammalian hosts against tick infestation has been proposed as the most sustainable alternative tick control method to the current use of acaricides which has several limitations. The success of this method is dependent on the identification of key molecules for use as tick vaccine antigens. Proteolytic enzymes are involved in a wide range of cellular processes in eukaryotes such as development regulation and nutrition, thus they can be considered as good target antigens for a tick vaccine. In the present study we used primers designed based on the consensus amino acid motifs flanking the conserved active sites C25 and N175 present in all papain-like cysteine proteinases to amplify by polymerase chain reaction, sequence and characterize two Haemaphysalis longicornis tick cysteine proteinase genes. Based on the nucleotide and deduced amino acid sequences, both genes were identified as members of the cysteine proteinase gene family by presence in their sequences of consensus motifs flanking the conserved active sites C25, H150 and N175 that are present in all papain-like cysteine proteinases. Both genes are about 1.2 kb in size and show high sequence homology predominantly to invertebrate cathepsin L like cysteine proteinases. PMID- 10379942 TI - Identification and structure of the Marek's disease virus serotype 2 glycoprotein M gene: comparison with glycoprotein M genes of Herpesviridae family. AB - We determined the nucleotide sequence of a portion of BamHI-C fragment of Marek's disease virus serotype 2 (MDV2) strain HPRS24 which was suspected to contain the homologue of the herpes simplex virus type 1 (HSV-1) gene UL10, encoding glycoprotein M (gM). An open reading frame whose translation product exhibited significant similarities to HSV-1 gM protein and respective proteins of other herpesviruses of 37.5% and 45.5% to 31.8%, respectively, was identified. A number of distinct transcriptional consensus sequences were found upstream of the first putative start codon of MDV2 UL10 protein. In transcriptional analysis, the gene was transcribed into an 1.5 kb RNA. The primary translation product comprises 424 amino acids with a predicted molecular weight of 46.9 kDa. The predicted MDV2 UL10 protein contains eight hydrophobic domains with sufficient length and hydrophobicity to span the lipid bilayer conserved in the genomes of all herpesviruses which have been sequenced so far. In the region located between the first and second hydrophobic domains, two potential N-linked glycosylation sites were presented. Interestingly, highly charged residues were abundantly possessed in the carboxy-terminal part of the MDV2 UL10 protein. By comparison of the amino acid sequence of the MDV2 UL10 gene with the homologues from other herpesviruses, the data might contribute for further evidence of the evolution of herpesviruses from a common progenitor and an ancient example of MDV2 belonging to the Alphaherpesvirinae subfamily. In addition, the existence of corresponding genes in human, mammalian, and avian herpesvirus genomes, suggests indirectly an important role for gM in the natural life cycle of the virus. PMID- 10379943 TI - Influence of motor activities on the release of transmitter quanta from motor nerve terminals in mice. AB - We investigated the effects of motor activities on transmitter release in mouse nerve-muscle preparations of the diaphragm muscle (DPH), extensor digitorum longus muscle (EDL), and soleus muscle (SOL). Mice were divided into a control group, a motor-restricted (RST) group, and a motor-compelled (CMP) group. The quantal content (m) of endplate potentials was measured intracellularly. In DPH the motor activity was unaffected. In the CMP group the m value of the EDL group increased with increases in the cooperativity of Ca2+ in transmitter release. Compared with the CMP group, the SOL of the RST group had a smaller m value with increases in the cooperativity of Ca2+ in transmitter release. These results suggest that motor activities can influence neuromuscular activity specific to different systems, however, the motor compulsion specifically activated the function of EDL and the motor restriction activated the function of SOL, and these effects might lead to altered activity of the release of transmitter quanta in motor nerve terminals of mice. PMID- 10379944 TI - The complete amino acid sequence of dog beta2-microglobulin. AB - Dog beta2-microglobulin was purified from the urine of dogs with potassium dichromate induced tubular damage. It was purified by sequential use of anion exchange chromatography, gel filtration chromatography, and reversed-phase high performance liquid chromatography. Comparisons of the amino acid sequence of the dog protein with human, mouse, and rabbit beta2-microglobulin, indicated a high degree of similarity. The dog protein was very similar to human beta2 microglobulin in that it had a molecular weight of 11.8 kDa and contained two half-cystinyl residues. Dog and human beta2-microglobulin were demonstrably different at 24 of the 99 positions compared. The data supported the conclusion that the purified protein was dog beta2-microglobulin and that all four proteins from dog, human, mouse, and rabbit were closely related. PMID- 10379945 TI - Plasma atrial and brain natriuretic peptide levels in dogs with congestive heart failure. AB - Plasma concentrations of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) were measured in 6 dogs with experimental mitral regurgitation (MR) and 19 canine patients with asymptomatic and symptomatic congestive heart failure (CHF). In dogs with experimental MR, ANP and BNP concentrations were significantly correlated with pulmonary capillary wedge pressure (PCWP) (ANP; r=0.852, P=0.0004, BNP; r=0.832, P=0.0008). ANP level was shown to have a predominant effect on PCWP in comparison with BNP using multiple regression analysis. In canine patients with asymptomatic and symptomatic CHF, ANP and BNP concentrations were significantly different among the heart failure classes according to the New York Heart Association functional classification (ANP; P=0.0165, BNP; P=0.0005). In addition, ANP and BNP levels in dogs with decompensated heart failure (n=10) significantly increased in comparison with those in dogs with compensated heart failure (n=9). There was however no correlation between ANP and BNP levels in each heart failure class. In conclusion, plasma ANP and BNP levels may become predictors of PCWP and the severity of heart failure in dogs with MR, although further investigations on ANP and BNP levels in more clinical cases are required. PMID- 10379946 TI - In vitro fertilization and cortical granule distribution of bovine oocytes having heterogeneous ooplasm with dark clusters. AB - In vitro maturation, fertilization and subsequent development of oocytes with homogeneous (category 1), or heterogeneous ooplasm (category 2) were investigated. No significant differences were observed in the nuclear maturation and total fertilization rates between the two categories. However, category 2 oocytes showed a higher normal fertilization rate due to their lower incidence of polyspermy as compared to category 1 oocytes. Electron microscopic study revealed that all category 2 oocytes had cortical granules lined up next to the plasma membrane, and that some category 1 oocytes still had small clusters of cortical granules after maturation. Although the proportion of cleaved zygotes was higher in category 2, the percentages of cleaved zygotes that developed to the blastocyst stage did not differ between the two categories. These results demonstrate that oocytes with heterogeneous ooplasm have a higher capacity for normal fertilization due to the reduction in polyspermy. This can be attributed to the normal distribution of cortical granules in category 2 oocytes after maturation. PMID- 10379947 TI - Morphological classification of ganglion cells in the central retina of chicks. AB - Classification of retinal ganglion cells (RGCs) in the chick central retina was studied by retrograde labeling of carbocyanine dye (DiI) and intracellular filling with Lucifer Yellow. Ganglion cells were divided into 4 groups, Group Ic/Is, Group IIc/IIs, Group IIIs, Group IVc, according to sizes of somal area and dendritic field and dendritic branching pattern. Group I cells had small somal area and small dendritic field. They were further divided into 2 subgroups by complexity (subgroup Ic) and simplicity (subgroup Is) of the dendritic arborization. Group II cells had medium-sized soma and dendritic field. They were also divided into subgroup IIc and IIs by the same definitions as those of subgroup Ic and Is. Group IIIs had medium-sized soma, large and simple dendritic arborization. Group IVc in which all cells had large soma, showed large and complex dendritic arborization. Cell populations of each group were 51.8% (subgroup Ic), 21.1% (subgroup Is), 6.2% (subgroup IIc), 14.6% (subgroup IIs), 4.2% (Group IIIs), and 2.1% (Group IVc). Subgroup Ic cells, which were very similar to beta-cells in the mammalian central area, represented about a half of the ganglion cell population. Cells in subgroup Is and IIs, which were not reported in the mammalian retina, were found in the chick central retina in relatively high population (35.7%). Morphological features of chick RGCs in the central retina were considered in comparison with those of other vertebrates. PMID- 10379948 TI - Age-related changes in rat hippocampal theta rhythms: a difference between type 1 and type 2 theta. AB - The age-related changes in two types of theta rhythms recorded from the hippocampus in young (4 months-old), mature (12-13 months-old) and aged (22-25 months-old) rats were investigated. The type 1 theta rhythm was measured from hippocampal EEG recorded from walking rats and the type 2 theta was measured from the EEG induced by reticular pontin oralis nucleus (PON) stimulation in urethane anesthetized rats. The peak frequency and the peak power were detected from power spectra calculated on each theta sample by fast Fourier transformation (FFT). No age-related alteration was observed on the peak frequency of type 1 theta rhythm. However, on type 2 theta rhythm, the peak frequency was decreased in the aged rats compared with the young and the mature rats. The type 2 theta rhythm is cholinergic, and therefore this result suggests that age-related deterioration can be clearly observed in the cholinergic system including the hippocampus in rats. PMID- 10379949 TI - Seroprevalence of bovine immunodeficiency virus in dairy and beef cattle herds in Korea. AB - Infection of bovine immunodeficiency virus (BIV), a lentivirus, is thought to sporadically occur throughout the world, but seroepidemiological surveys concerning the incidence of BIV are limited and have not been undertaken in Korea. A total of 266 sera from different twenty dairy (Holstein) and twenty-six Korean native beef (Hanwoo) farms of the south-western part of Korea was analyzed for the presence of anti-BIV antibodies by Western blotting. Thirty five percent and 33% of dairy and beef cattle, respectively, were BIV-seropositive. By nested polymerase chain reaction, it was confirmed that these seropositive cows had provirus in the peripheral blood mononuclear cells. To demonstrate the correlation with BIV and bovine leukemia virus (BLV) infection, these sera were also analyzed for anti-BLV antibodies by immunodiffusion test, resulting in high prevalence of BLV infection but relatively a few dual infections. We report herein the first serological detection of antibodies to BIV in Korea. PMID- 10379950 TI - Effects of isoprothiolane on cell growth of cultured bovine mammary epithelial cells. AB - This study was performed to investigate the effects of isoprothiolane on cell growth and the production of interleukin (IL)-1 and IL-6 by bovine mammary epithelial cells in vitro. Isoprothiolane increased proliferation of mammary epithelial cells in a dose-dependent manner at the concentration of 0.05 to 5 microM when cultured either with or without serum-supplemented medium. In contrast, isoprothiolane (0.0005-5 microM) significantly inhibited the production of IL-1 and IL-6 by mammary epithelial cells. Moreover, the cytokines, IL-1alpha, IL-1beta, IL-6, and tumor necrosis factor (TNF)-alpha tended to inhibit the proliferation of mammary epithelial cells in a dose-dependent manner. These results indicated that isoprothiolane regulated mammary epithelial cell growth in vitro possibly by modulating the production of cytokines. PMID- 10379951 TI - Peripheral nerve lesions in a case of equine motor neuron disease. AB - A male 14-year-old Arab horse was pathologically diagnosed as equine motor neuron disease (EMND), which was kept as a breeding horse on a farm in Tokachi district of Hokkaido in Japan. On examination of the peripheral nerves, the most characteristic feature was Wallerian-type degeneration revealed by myelinoclasis associated with myelin ovoids which were sometimes infiltrated by macrophages. The other abnormalities were axonal swellings which were surrounded by thin myelin sheaths. Ultrastructurally, the axonal swelling was due to an accumulation of neurofilaments, and was accompanied by a thin and degenerating myelin sheaths. In teased nerve fiber preparations, the most conspicuous change was myelinoclasis represented by segmentation into myelin ovoids or balls. Occasionally, segmental demyelination and axonal degeneration characterized by multifocal axonal swelling were observed. PMID- 10379952 TI - Thymic carcinoma of the thymic hormone secretory type in a cow. AB - An 8-year-old Holstein cow had tumor nodules and enlarged lymph nodes in the mediastinum, and metastatic tumor masses in the pelvic cavity. The neoplastic cells were characterized by squamous features and intracytoplasmic vacuoles carrying microvilli, some of which contained periodic acid Schiff-positive globular cores, but tubular structures or goblet cells were absent. Many neoplastic cells stained positively for keratin, and occasional cells were positive for thymosin. The presence of secretory granules in the cytoplasm was confirmed by electron microscopy. This neoplasm was considered to be of thymic hormone-secreting epithelial cell origin. PMID- 10379953 TI - Effects of the lipopolysaccharide-protein complex and crude capsular antigens of Pasteurella multocida serotype A on antibody responses and delayed type hypersensitivity responses in the chicken. AB - The effects of the lipopolysaccharide-protein complex (LPS) and crude capsular antigen (CCA) prepared from Pasteurella multocida serotype A isolated from a duck in the Philippines, on antibody responses to sheep red blood cells (SRBC) and Brucella abortus (BA) and delayed type hypersensitivity (DTH) responses to bovine serum albumin (BSA) in the chickens were studied. Chickens injected subcutaneously with LPS and CCA at 1 and 2 weeks of age and immunized intravenously with the mixed antigens of SRBC and BA, at 3 and 4 weeks of age showed significantly increased antibody responses against both SRBC and BA, when evaluated at 7 days after each immunization. In addition, these chickens sensitized intramuscularly with the emulsion of BSA in complete Freund's adjuvant at 5 weeks of age, and then injected into the wattle with BSA at 7 weeks of age also showed significantly increased DTH responses against BSA, when evaluated at 24 and 48 hr after challenge. These results indicate that LPS and CCA of P. multocida serotype A have a property enhancing humoral and cell-mediated immune responses. PMID- 10379954 TI - Effect of vitamin B2 on somatic cell counts in milk of clinical Staphylococcus aureus mastitis. AB - Effects of intravenous injection of Vitamin B2 (VB2) on the nitroblue tetrazolium (NBT) reductivity of peripheral blood neutrophils and the somatic cell counts (SCC) in quarter milk of Staphylococcus aureus mastitis were investigated. The NBT reductivities of neutrophils were enhanced at 2 days after single injection of VB2 (5.0 and 2.5 mg/kg), and were also enhanced at 4 days after initial injection of continuous 3 days of VB2 (2.5 mg/kg). The SCC in quarter milk were significantly decreased at 3, 7 and 14 days after initial injection of continuous 3 days of VB2 (2.5 mg/kg), however, S. aureus in the infected quarter was not cured bacteriologically by VB2 injection. PMID- 10379955 TI - The fungicide carbendazim induces meiotic micronuclei in the spermatids of the rat testis. AB - Whether the fungicide carbendazim affects the meiotic spermatocytes and consequently induces chromosome aberrations in the spermatids was determined in the adult rat testis using the micronucleus test. Round spermatids containing micronuclei (MN) were significantly increased in number at stages I and V on days 1 and 4.5 after treatment with carbendazim (100 mg/kg), respectively (p<0.05). Immunocytochemistry indicated that approximately 68% of the carbendazim-induced MN contained kinetochores. These results suggest that carbendazim induces chromosome aberrations in spermatids with a high incidence of aneuploidy. PMID- 10379956 TI - Differentiated embryonal rhabdomyosarcoma in a cow. AB - An embryonal rhabdomyosarcoma was found in the pleura of a 2-year-old Holstein cow after first delivery. The most predominant cells in the tumor were relatively small in size, but considerable numbers of more differentiated cells of larger sizes mingled with the small cells. The most differentiated cells were characterized by multinucleation, abundant cytoplasm containing cross-striated fibrils, intense immunoreactivity for desmin, and weak or negative reactivity for vimentin. Such cells, lacking mitotic activity and displaying weak or no reactivity for proliferating cell nuclear antigen, were considered to be malignant counterparts of myotubes or muscle fibers. This neoplasm seems to follow normal skeletal muscle embryogenesis, and to be capable of differentiation into the final stage of muscle development. PMID- 10379957 TI - Aberrant expression of retinoid receptors and lung carcinogenesis. PMID- 10379958 TI - High-dose chemo and breast cancer: early results spark debate. PMID- 10379959 TI - Breast cancer increases on Papua New Guinea. PMID- 10379960 TI - Managed care "de-skills" the oncology nurse, ONS says. Oncology Nursing Society. PMID- 10379962 TI - Traditional Chinese medicine taps "qi" for health. PMID- 10379961 TI - Second line therapies move to the forefront in colorectal cancer. PMID- 10379963 TI - Analyzing health surveys for cancer-related objectives. AB - Large-scale health surveys conducted by government agencies record information on a large number of health-related variables. We review the use of these data for performing analyses that address cancer-related objectives. After describing the conduct of a large-scale health survey (the third National Health and Nutrition Examination Survey [NHANES III]), we discuss some of the issues involved in analyzing data collected in such a survey. In particular, the use of sample weights in the analysis and the importance of accounting for the complex survey design when estimating standard errors are discussed. Six applications are then presented that involve the following: 1) estimating demographic factors associated with snuff use, 2) estimating the association of type of health insurance with the probability of receiving a digital rectal examination, 3) estimating the association of body iron stores with the probability of later developing cancer, 4) estimating the changing rates of mammography screening in the United States between 1987 and 1992, 5) evaluating smoking and alcohol consumption as risk factors for digestive cancer by use of a population-based, case-control study, and 6) evaluating a randomized community-intervention experiment to encourage smoking cessation. These applications use data from the National Health Interview Survey, the NHANES I Epidemiologic Followup Study, the 1986 National Mortality Followback Survey, and the Community Intervention Trial for Smoking Cessation. The availability of public-use data files is discussed for surveys sponsored by the U.S. government that collect health-related information. We demonstrate that statistical methods and computer software are available for analyzing public-use data files of surveys to address different types of cancer related objectives. PMID- 10379964 TI - Cancer surveillance series: interpreting trends in prostate cancer--part I: Evidence of the effects of screening in recent prostate cancer incidence, mortality, and survival rates. AB - BACKGROUND: The prostate-specific antigen test was approved by the U.S. Food and Drug Administration in 1986 to monitor the disease status in patients with prostate cancer and, in 1994, to aid in prostate cancer detection. However, after 1986, the test was performed on many men who had not been previously diagnosed with prostate cancer, apparently resulting in the diagnosis of a substantial number of early tumors. Our purpose is to provide insight into the effect of screening on prostate cancer rates. Detailed data are presented for whites because the size of the population allows for calculating statistically reliable rates; however, similar overall trends are seen for African-Americans and other races. METHODS: Prostate cancer incidence data from the National Cancer Institute's Surveillance, Epidemiology, and End Results Program and mortality data from the National Center for Health Statistics were analyzed. RESULTS/CONCLUSIONS: The following findings are consistent with a screening effect: 1) the recent decrease since 1991 in the incidence of distant stage disease, after not having been perturbed by screening; 2) the decline in the incidence of earlier stage disease beginning the following year (i.e., 1992); 3) the recent increases and decreases in prostate cancer incidence and mortality by age that appear to indicate a calendar period effect; and 4) trends in the incidence of distant stage disease by tumor grade and trends in the survival of patients with distant stage disease by calendar year that provide suggestive evidence of the tendency of screening to detect slower growing tumors. IMPLICATIONS: The decline in the incidence of distant stage disease holds the promise that testing for prostate-specific antigen may lead to a sustained decline in prostate cancer mortality. However, population data are complex, and it is difficult to confidently attribute relatively small changes in mortality to any one cause. PMID- 10379965 TI - Cancer surveillance series: interpreting trends in prostate cancer--part II: Cause of death misclassification and the recent rise and fall in prostate cancer mortality. AB - BACKGROUND: The rise and fall of prostate cancer mortality correspond closely to the rise and fall of newly diagnosed cases. To understand this phenomenon, we explored the role that screening, treatment, iatrogenic (i.e., treatment-induced) deaths, and attribution bias (incorrect labeling of death from other causes as death from prostate cancer) have played in recent mortality trends. METHODS: Join point regression is utilized to assess the recent rise and fall in mortality and the relationship of total U.S. trends to those areas served by the National Cancer Institute's Surveillance, Epidemiology, and End Results (SEER) Cancer Registry Program. Incidence-based mortality (IBM) is estimated with the use of prostate cancer data from the SEER Program to partition (from overall prostate cancer mortality trends) the contribution of cases diagnosed since the widespread use of prostate-specific antigen (PSA) testing starting in 1987. IBM is also used to examine the contribution of stage at diagnosis to the recent prostate cancer mortality trends. RESULTS: IBM for cases diagnosed since 1987 rose above the pre 1987 secular (i.e., background) trend, peaked in the early 1990s, and almost returned to the secular trend by 1994. This rise and fall of IBM track with the pool of prevalent cases diagnosed within the prior 2 years. IBM for cases diagnosed with metastatic disease fell starting in 1991, while IBM for those diagnosed with localized/regional disease was relatively flat. CONCLUSIONS: The rise and fall in prostate cancer mortality observed since the introduction of PSA testing in the general population are consistent with a hypothesis that a fixed percent of the rising and falling pool of recently diagnosed patients who die of other causes may be mislabeled as dying of prostate cancer. The decline in IBM for distant stage disease and flat IBM trends for localized/regional disease provide some evidence of improved prognosis for screen-detected cases, although alternative interpretations are possible. PMID- 10379966 TI - Cancer surveillance series: interpreting trends in prostate cancer--part III: Quantifying the link between population prostate-specific antigen testing and recent declines in prostate cancer mortality. AB - BACKGROUND: The objective of this study was to investigate the circumstances under which dissemination of prostate-specific antigen (PSA) testing, beginning in 1988, could plausibly explain the declines in prostate cancer mortality observed from 1992 through 1994. METHODS: We developed a computer simulation model by use of information on population-based PSA testing patterns, cancer detection rates, average lead time (the time by which diagnosis is advanced by screening), and projected decreased risk of death associated with early diagnosis of prostate cancer through PSA testing. The model provides estimates of the number of deaths prevented by PSA testing for the 7-year period from 1988 through 1994 and projects what prostate cancer mortality for these years would have been in the absence of PSA testing. RESULTS: Results were generated by assuming a level of screening efficacy similar to that hypothesized for the Prostate, Lung, Colorectal, and Ovarian (PLCO) Cancer Screening Trial. Under this assumption, the projected mortality in the absence of PSA testing continued the increasing trend observed before 1991 only when it was assumed that the mean lead time was 3 years or less. Projected mortality trends in the absence of PSA screening were not consistent with pre-1991 increasing trends for lead times of 5 years and 7 years. CONCLUSIONS: When screening is assumed to be at least as efficacious as hypothesized in the PLCO trial, it is unlikely that the entire decline in prostate cancer mortality can be explained by PSA testing based on current beliefs concerning lead time. Only very short lead times would produce a decline in mortality of the magnitude that has been observed. PMID- 10379967 TI - Cancer surveillance series: changing geographic patterns of lung cancer mortality in the United States, 1950 through 1994. AB - BACKGROUND: Geographic surveys revealing variations in lung cancer mortality rates across the United States have prompted epidemiologic studies in high-risk communities. We have updated these maps to track the changing patterns and to provide further clues to the determinants of lung cancer. METHODS: Age-adjusted race- and sex-specific lung cancer mortality rates from 1950 through 1994 were calculated for nine Census Divisions and 508 State Economic Areas of the United States. RESULTS: Pronounced geographic variation in lung cancer rates was evident, with the patterns changing substantially over time. Among white males in the 1950s and 1960s, high rates were observed in urban areas of the northeast and north central states and in areas along the southeast and Gulf coasts. By the 1970s, the northern excess began to fade, with high rates starting to cover wider areas of the south. By the 1980s to the mid-1990s, clustering of elevated rates was prominent across the southeast and south central areas, with relatively low rates throughout much of the northeast. Among white females, little geographic variation was evident in the 1950s, but thereafter relatively high rates began to appear in clusters along the Atlantic and Pacific coasts. For both sexes, consistently low rates were seen in the mountain and the plains states. Rates among blacks were consistently elevated in northern areas and low across the south. CONCLUSIONS: The changing mortality patterns for lung cancer generally coincide with regional trends in cigarette smoking, indicating that public health measures aimed at smoking prevention and cessation should have a dramatic effect in reducing lung cancer rates. PMID- 10379968 TI - Cancer surveillance series: recent trends in childhood cancer incidence and mortality in the United States. AB - BACKGROUND: Public concern about possible increases in childhood cancer incidence in the United States led us to examine recent incidence and mortality patterns. METHODS: Cancers diagnosed in 14540 children under age 15 years from 1975 through 1995 and reported to nine population-based registries in the National Cancer Institute's Surveillance, Epidemiology, and End Results Program were investigated. Age-adjusted incidence was analyzed according to anatomic site and histologic categories of the International Classification of Childhood Cancer. Age-adjusted U.S. mortality rates were calculated. Trends in rates were evaluated by use of standard regression methods. RESULTS: A modest rise in the incidence of leukemia, the most common childhood cancer, was largely due to an abrupt increase from 1983 to 1984; rates have decreased slightly since 1989. For brain and other central nervous system (CNS) cancers, incidence rose modestly, although statistically significantly (two-sided P = .020), largely from 1983 through 1986. A few rare childhood cancers demonstrated upward trends (e.g., the 40% of skin cancers designated as dermatofibrosarcomas, adrenal neuroblastomas, and retinoblastomas, the latter two in infants only). In contrast, incidence decreased modestly but statistically significantly for Hodgkin's disease (two sided P = .037). Mortality rates declined steadily for all major childhood cancer categories, although less rapidly for brain/CNS cancers. CONCLUSIONS: There was no substantial change in incidence for the major pediatric cancers, and rates have remained relatively stable since the mid-1980s. The modest increases that were observed for brain/CNS cancers, leukemia, and infant neuroblastoma were confined to the mid-1980s. The patterns suggest that the increases likely reflected diagnostic improvements or reporting changes. Dramatic declines in childhood cancer mortality represent treatment-related improvements in survival. PMID- 10379969 TI - Expression of retinoid receptor genes and proteins in non-small-cell lung cancer. AB - BACKGROUND: Retinoids can suppress carcinogenesis in high-risk non-neoplastic bronchial lesions and can reduce the risk of second primary non-small-cell lung cancer (NSCLC). The effects of retinoids are mediated by nuclear receptors, i.e., the retinoic acid receptors (RARalpha, RARbeta, and RARgamma) and the retinoid X receptors (RXRalpha, RXRbeta, and RXRgamma). We investigated whether abnormalities in the in vivo expression of retinoid receptors are observed in NSCLC. METHODS: Expression of retinoid receptors in paired specimens of normal and cancerous tissues from the lungs of 76 patients with NSCLC was studied by use of antiretinoid receptor antibodies (except those against RXRgamma) and immunohistochemistry. RAR messenger RNAs were analyzed by use of in situ hybridization and by reverse transcription-polymerase chain reaction (RT-PCR). Samples were also studied for loss of heterozygosity (LOH) at chromosome 3p24. All P values are two-sided. RESULTS: All studied receptors were expressed in normal lung cells and in high- risk non-neoplastic lesions. In tumor cells, overexpression of RXRalpha and RARalpha was frequently observed. In contrast, RXRbeta expression decreased in 18% of the tumor specimens. Furthermore, there was a marked decrease in the expression of RARbeta in 63% of the tumors (P<.0001). Decreased expression of RARgamma was observed by RT-PCR in 41% of the tumors (P<.0001). LOH at 3p24 was observed in 41% of the tumor specimens from informative patients and in 20% of the non-neoplastic lesions. CONCLUSIONS: Expression of RARalpha and RXRalpha is either normal or elevated in NSCLC. In contrast, a large percentage of tumors show a marked decrease in the expression of RARbeta, RARgamma, and RXRbeta as well as a high frequency of LOH at 3p24, which was also observed in non-neoplastic lesions. These data suggest that altered retinoid receptor expression may play a role in lung carcinogenesis. PMID- 10379970 TI - Urinary 2-hydroxyestrone/16alpha-hydroxyestrone ratio and risk of breast cancer in postmenopausal women. AB - BACKGROUND: It has been suggested that women who metabolize a larger proportion of their endogenous estrogen via the 16alpha-hydroxylation pathway may be at elevated risk of breast cancer compared with women who metabolize proportionally more estrogen via the 2-hydroxylation pathway. However, the supporting epidemiologic data are scant. Consequently, we compared the ratio of urinary 2 hydroxyestrone (2-OHE1) to 16alphahydroxyestrone (16alpha-OHE1) in postmenopausal women with breast cancer and in healthy control subjects. METHODS: Estrogen metabolites were measured in urine samples obtained from white women who had participated in a previous population-based, breast cancer case-control study at our institution. All P values are from two-sided tests. RESULTS: All of the urinary estrogens measured, with the exception of estriol, were higher in the 66 case patients than in the 76 control subjects. The mean value of urinary 2-OHE1 in case patients was 13.8% (P = .20) higher than that in control subjects, 16alpha-OHE1 was 12.1% (P = .23) higher, estrone was 20.9% higher (P = .14), and 17beta-estradiol was 12.0% higher (P = .36). The ratio of 2-OHE1 to 16alpha-OHE1 was 1.1% higher in the patients (P = .84), contrary to the hypothesis. Compared with women in the lowest third of the values for the ratio of urinary 2-OHE1 to 16alpha-OHE1, women in the highest third were at a nonstatistically significantly increased risk of breast cancer (odds ratio = 1.13; 95% confidence interval = 0.46-2.78), again contrary to the hypothesis. CONCLUSION: This study does not support the hypothesis that the ratio of the two hydroxylated metabolites (2 OHE1/16alpha-OHE1) is an important risk factor for breast cancer. PMID- 10379971 TI - Re: Statewide study of diagnostic agreement in breast pathology. PMID- 10379972 TI - Continuing: Biology of cachexia. PMID- 10379973 TI - Continuing: Biology of cachexia. PMID- 10379974 TI - Re: Sunscreen use, wearing clothes, and number of nevi in 6- to 7-year-old European children. PMID- 10379975 TI - Re: Chronically depressed mood and cancer risk in older persons. PMID- 10379976 TI - Revisiting: Involuntary smoking and lung cancer: a case-control study. PMID- 10379977 TI - FRACAS: a system for computer-aided image-guided long bone fracture surgery. AB - This article describes FRACAS, a computer-integrated orthopedic system for assisting surgeons in performing closed medullary nailing of long bone fractures. FRACAS's goal is to reduce the surgeon's cumulative exposure to radiation and surgical complications associated with alignment and positioning errors of bone fragments, nail insertion, and distal screw locking. It replaces uncorrelated, static fluoroscopic images with a virtual reality display of three-dimensional bone models created from preoperative computed tomography and tracked intraoperatively in real time. Fluoroscopic images are used to register the bone models to the intraoperative situation and to verify that the registration is maintained. This article describes the system concept, software prototypes of preoperative modules (modeling, nail selection, and visualization), intraoperative modules (fluoroscopic image processing and tracking), and preliminary in vitro experimental results to date. Our experiments suggest that the modeling, nail selection, and visualization modules yield adequate results and that fluoroscopic image processing with submillimetric accuracy is practically feasible on clinical images. PMID- 10379978 TI - Iliosacral screw insertion using computer-assisted CT image guidance: a laboratory study. AB - Procedures were performed to evaluate the efficacy of using a computer-assisted image-guided system to improve the accuracy of inserting iliosacral screws for posterior pelvic ring injuries. The fluoroscopic method currently in use has shortcomings that do not eliminate the risk to the L5 and sacral nerve roots and iliac vessels. The procedure was performed on embalmed cadaver pelvis specimens with intact soft tissues. Iliosacral screws were inserted across each sacroiliac joint after registering the opposite ilium to simulate a clinical situation. Two trials were performed. The first was necessary to become familiar with the system and test the basic methods of registration. In the second trial, deficiencies were corrected by using spherical-headed fiducial screws for fiducials and a better reference frame clamp, and by better spacing of more fiducials. Results were evaluated by inlet and outlet X-ray views and dissection. All eight S-1 iliosacral 7.0-mm cannulated screws were entirely inside bone. Of the eight S-2 screws, five were inside bone, two infringed upon an S-1 foramen, and one had threads out of the sacral body anteriorly and intruding into an S-1 foramen. This system was felt to be effective and sufficiently safe to warrant clinical trials. PMID- 10379979 TI - Computer-assisted spine surgery. AB - The aim of this study was to improve the reliability of pedicle screw insertion. Transpedicle screw insertion may cause neurological, vascular, and mechanical complications. Previous studies of surgical procedures have shown a significant rate of incorrect placement of the screw ranging from 10 to 40%. A new technique that combines preoperative computed tomography (CT) imaging with intraoperative passive navigation was used to perform 64 pedicle screw insertions in the thoracolumbar region. At the same time, 64 pedicle screw insertions were performed manually in the same region and on the same vertebral levels. Surgery was followed in all cases by postoperative radiographs and computed tomography examination, which allowed measurements of screw position relative to pedicle position to be performed. A comparison between the two groups showed that six screws in 64 vertebra (9%) had incorrect placement with the computer-assisted technique whereas 28 screws in 64 vertebra (44%) had incorrect placement with manual insertion. The intraoperative accuracy provided by the computer after registration was better than 1 mm. The good results obtained are similar to those reported in the literature. The cortex penetration observed with the computer assisted technique was not imputed to computer failure. Errors by the surgeon in acquiring data in the pre- and perioperative steps may explain the six incorrect screw placements. This clinical experience confirms that the accuracy and the reliability of this computer-assisted technique are good. PMID- 10379980 TI - Experiences in intraoperative computer-aided navigation in ENT sinus surgery with the Aesculap navigation system. AB - Five patients with chronic sinus pathology and an indication for sinus surgery were selected. For intraoperative navigation, we used Surgical Planning and Orientation Computer Systems (SPOCS) Aesculap navigation software (ISG Technologies, Mississauga, Ontario, Canada) and surgical instruments fitted with light-emitting diodes. Navigation procedures are described in detail in the article. The system's precision was measured by pointing at anatomical landmarks. The accuracy was measured as the distance in millimeters between the bony structures of the computed tomographic (CT) scan on screen and the cross-hair of the pointer tip displayed on the screen. Another parameter of the system's accuracy was calculated by the system itself as the root mean square error in millimeters between the markers' position as registered and their position in the CT data set. Axial 3/3/1-mm spiral CT provided sufficient resolution, and data transfer via optical disk was practicable. Positioning of the navigation equipment required some experience, and the registration of the patient's head position also needed attention, as the markers have to be pointed at precisely. During the operation, the position of the head-tracking system on the patient's head must remain unchanged to ensure a correct navigation display. The main advantage of the computed navigation system was the constant orientation provided during the sinus surgical procedure. Borders and critical anatomical structures could be identified in the corresponding CT data set, thus enabling the surgeon to decide on subsequent procedures. Use of the navigation system was found to increase the operation time by about 1 h, resulting in additional time under anesthesia. We found the SPOCS Aesculap computed navigation system to be an established technical aid, ready for use in ENT sinus surgery. In the cases reported here, a precision between 1 and 3 mm was obtained. PMID- 10379981 TI - Surgical management of intraosseous skull base tumors with aid of Operating Arm System. AB - Invasion of bone and critical neurovascular structures often impedes complete resection of intraosseous skull base neoplasms, and these lesions tend to recur unless all infiltrated bone is removed. Evolving experience with image guidance over the past few years indicates the potential value of neuronavigation in skull base lesions diffusely infiltrating or fixed to bone structures. We report our early experience with the Radionics Operating Arm System (OAS), specifically emphasizing its utility as an adjunct in the treatment of intraosseous skull base tumors, mainly meningiomas. In April 1995 the OAS was introduced into clinical use at the neurosurgical university clinic in Munster, Germany. Since then, the system's utility has been explored in 10 patients out of the total neuronavigation series presenting with intraosseous skull base tumors (nine females and one male, mean age 47 years; nine meningiomas, one chordoma). For navigational planning, both 3-mm computed tomography scans and a set of 3-mm fat suppression magnetic resonance images were chosen. At least four adhesive skin markers were used for system calibration. The system was technically usable in all cases in this small series. Because of the relative immobility of the bone structures and/or the tumor, no significant deviation from the preoperative registration accuracy was noted at the end of the procedures. The main advantages were easier localization and resection of infiltrated bone, which is often not grossly identifiable, even under the microscope. Our preliminary experience with the OAS suggests that image guidance is helpful in this type of lesion, providing better anatomical orientation during surgery and delineating tumor margins and their relation to critical neurovascular structures. The problem of a possible intracranial tumor and brain shift can be neglected in these lesions. The system facilitates resection by volumetric contour information, allowing more aggressive and complete resection. PMID- 10379982 TI - Computed-tomography-based computer preoperative planning for total hip arthroplasty. AB - For precise preoperative planning in total hip arthroplasty (THA), we developed a technique of computed tomography (CT)-based computer preoperative planning and compared this technique with the single X-ray and template method generally used. The subjects of this study were 42 hips in 38 patients who underwent THA using a cementless total hip system. Preoperatively, a standard anteroposterior X-ray of the hip was taken, and conventional preoperative planning was done with a template of the total hip system. Transverse images were obtained using a helical CT scanner, and a CT-based computer preoperative plan was performed on true coronal slice images of the proximal femur reconstructed from CT data. Postoperatively, 29 hips (69%) showed good proximal fit of the femoral component to the medial endosteal line. Of the 20 hips with good proximal fit on preoperative X-ray planning, 12 hips had good proximal fit on postoperative X rays. Sensitivity and specificity of the proximal fit on X-ray templating were 41 and 23%, respectively. In 27 of 28 hips with good proximal fit on reconstructed CT images preoperatively, the postoperative X ray revealed good proximal fit. Sensitivity and specificity of the proximal fit on computer planning were 93 and 86%, respectively. Twelve hips with good proximal fit on preoperative templating, the reconstructed images, and the postoperative X ray had 20 degrees or less of combined femoral neck anteversion and external rotational contracture of the hip on the X-ray table. Eight hips with good proximal fit on preoperative templating and proximal poor fit on the reconstructed images had 17-65 degrees of combined version and rotational contracture. In 16 hips with poor proximal fit on preoperative templating and good proximal fit on the reconstructed images, the combined version and rotational contracture ranged from 17 to 69 degrees. When combined femoral neck anteversion and external rotational contracture of the hip is less than 15 degrees, the simple X-ray and template method might be sufficient for THA planning. Otherwise, the CT-based method of preoperative planning is recommended. PMID- 10379983 TI - Hormonal regulation of microsomal cytochrome P4502E1 and P450 reductase in rat liver and kidney. AB - 1. The relative roles of pituitary hormones (especially growth hormone) and testicular hormones (especially testosterone) in the regulation of renal and hepatic CYP2E1 and cytochrome P450 reductase have been studied in the male rat. 2. Depletion of pituitary hormones by hypophysectomy (Hx) resulted in 12-14-fold increases in renal CYP2E1 (p < or = 0.05) and a 40% drop in NADPH-dependent cytochrome c reductase activity (p < or = 0.05) compared with 6-fold increases in CYP2E1 (p < or = 0.05) and a 60% drop in P450 reductase apoprotein (p < or = 0.05) in the liver. 3. The increase in hepatic CYP2E1 was associated with increased gene transcription in nuclear run-on experiments. 4. Restoration of renal CYP2E1 to control levels by hormone treatment required both growth hormone and an intact testis, whereas partial restoration of CYP2E1 apoprotein levels in liver was accomplished by growth hormone, but not testosterone. 5. Renal NADPH dependent cytochrome c reductase activity was restored by growth hormone and testosterone treatment, whereas the hepatic reductase appeared to be regulated by other pituitary hormones. 6. CYP2E1 and P450 reductase appear to be under complex endocrine regulation by pituitary and testicular hormones in a tissue specific manner. PMID- 10379984 TI - Biotransformation of the antipsychotic agent, mazapertine, in dog--mass spectral characterization and identification of metabolites. AB - 1. Biotransformation of the antipsychotic agent, mazapertine, was studied after a single oral administration of 14C-mazapertine succinate (10 mg/kg, free base) to six beagle dogs (three male, three female). 2. Following oral administration of 14C-mazapertine, plasma (0-48 h), urine (0-7 days), and faeces (0-7 days) were collected. Recoveries of total radioactivity in urine and faeces were 26.9 and 62.0% of the dose, respectively. 3. Unchanged mazapertine plus 14 metabolites were isolated and identified, which accounted for > 60% of the sample radioactivity in the plasma, 17% of the dose in urine and 28% of the dose in faecal extract. 4. Unchanged mazapertine accounted for < 4% of the radioactive dose in excreta samples and < 21% of the sample radioactivity present in plasma samples. 5. Seven metabolic pathways for the formation of metabolites were identified including: (1) phenyl hydroxylation, (2) piperidyl oxidation, (3) O dealkylation, (4) N-dephenylation, (5) oxidative N-debenzylation, (6) depiperidylation and (7) conjugation. 6. Pathways 1, 2, 5 and 6 produced 4-OH piperidyl, OH-phenyl-OH-piperidyl, carboxybenzoyl piperidine and depiperidyl analogues of mazapertine as major metabolites. PMID- 10379985 TI - A comparison of basal and induced hepatic microsomal cytochrome P450 monooxygenase activities in the cynomolgus monkey (Macaca fascicularis) and man. AB - 1. The specific activities of hepatic microsomal cortisol 6beta-hydroxylase, coumarin 7-hydroxylase, S-mephenytoin 4'-hydroxylase and phenoxazone hydroxylase and the O-dealkylations of seven homologous alkoxyresorufins were < 3-fold different between the untreated (UT) cynomolgus monkey and man. 2. Heptoxy- and octoxyresorufin O-dealkylase, S-mephenytoin N-demethylase and dextromethorphan O demethylase specific activities were > 6-fold higher, whereas tolbutamide hydroxylase was almost 5-fold lower in the UT monkey than in man. 3. Phenobarbitone induced (2-6-fold) coumarin 7-hydroxylase, cortisol 6beta hydroxylase, S-mephenytoin N-demethylase, phenoxazone hydroxylase and benzyloxyresorufin O-dealkylase activities, but not the O-dealkylations of pentoxyresorufin or other alkoxyresorufins, in monkey. 4. Rifampicin induced (2-3 fold) cortisol 6beta-hydroxylase, S-mephenytoin 4'-hydroxylase, S-mephenytoin N demethylase and tolbutamide hydroxylase activities, the O-dealkylations of methoxy-, ethoxy- and propoxyresorufin and CYP2C- and CYP3A-immunorelated proteins in monkey. 5. Dextromethorphan O-demethylase was significantly reduced by both phenobarbitone and rifampicin treatment in monkey. 6. Beta-naphthoflavone induced (8-39-fold) the O-dealkylations of several alkoxyresorufins, the greatest effect being on propoxyresorufin, but had no effect on the other activities measured in monkey. 7. Constitutive hepatic microsomal CYP2D6-immunorelated proteins were expressed at apparently much higher levels in monkey than in man. PMID- 10379987 TI - Biotransformation of pentachlorophenol, aniline and biphenyl in isolated rainbow trout (Oncorhynchus mykiss) hepatocytes: comparison with in vivo metabolism. AB - 1. The biotransformation of pentachlorophenol (PCP), aniline and biphenyl in rainbow trout (Oncorhynchus mykiss) isolated liver cells was investigated to examine if fish hepatocytes represent a suitable alternative to the in vivo approach for studying the biotransformation of chemicals. Each compound was incubated at two concentrations (10 and 60 microM) for 2 h. For comparison, the metabolic profile of these xenobiotics was also studied in urine and bile of trout orally exposed to 1.8-4.0 mg/kg wet wt of each compound. 2. In vitro as in vivo, PCP glucuronide and to a lesser extent PCP sulphate were the metabolites formed by trout from PCP. 3. Aniline was mainly metabolized to acetanilide and to a lesser extent to 2-aminophenol by isolated hepatocytes, but neither hydroxylated acetanilide nor conjugates were found in vitro whereas they were present in bile and urine of trout treated with this chemical. 4. Trout hepatocytes metabolized biphenyl to hydroxylated and dihydroxylated products and the corresponding glucuronides. These results correlated well with the metabolic profile obtained from the bile of trout exposed to this pesticide. 5. It is concluded that although hepatocytes are well suited for several types of biotransformation studies, the fact that this system may in some cases produce a different metabolic pattern than in vivo should be considered when attempting to extrapolate in vitro to in vivo data. PMID- 10379986 TI - Beta2-agonist abuse in food producing animals: use of in vitro liver preparations to assess biotransformation and potential target residues for surveillance. AB - 1. The biotransformation of [3H]clenbuterol, [3H]salbutamol, [14C]salmeterol and 7-ethoxycoumarin by bovine liver was investigated by incubation with freshly prepared microsomes, suspension and monolayer cultures of isolated hepatocytes, precision-cut (250 microm) and chopped (600 microm) tissue slices. 2. Radio-HPLC analysis indicated that the saligenin beta2-agonists salmeterol and salbutamol were extensively metabolized by all intact cell preparations. A single major product (SmM1) was evident for salmeterol and two unresolved products for salbutamol (SbM1 and SbM2). Differential enzyme hydrolysis studies with Helix pomatia beta-glucuronidase/aryl sulphatase indicated that the main metabolites were glucuronide conjugates. Consistent with this, analysis of metabolites by liquid chromatography-mass spectrometry showed molecular ions ([M+H]+) at m/z 592 for Sm1 and 416 for both Sb1 and Sb2. 3. Comparable studies with clenbuterol revealed three minor metabolites. Prolonged incubations generated products representing, at maximum, 27% biotransformation. Two of the products have been identified as a glucuronide ([M+H]+, m/z 453) and hydroxyclenbuterol ([M+H]+, m/z 293). 4. These findings indicate that in vitro studies provide simple and cost effective means of evaluating xenobiotic metabolism, and thus of identifying potential target residues to enable surveillance of use of unlicensed veterinary drugs, or prohibited substances in farm animals. PMID- 10379989 TI - Haloalcohols deplete glutathione when incubated with fortified liver fractions. AB - 1. This study has examined the ability of dichloropropanols, haloalcohols and their putative metabolites to deplete glutathione when incubated with liver fractions obtained from untreated and differentially induced rats. 2. 1,3 Dichloropropan-2-ol and 2,3-dichloropropan-1-ol (0-1000 microM) both depleted glutathione in a dose-dependent manner when incubated with cofactors (NADPH generating system) and liver microsomes from the untreated rat. 3. The extent of GSH depletion was significantly enhanced when liver microsomes from the isoniazid or isosafrole-treated rat were used. 4. Epichlorohydrin produced a moderate, dose-dependent depletion of GSH. By contrast, 1,3-dichloroacetone (identified by TLC as a metabolite of 1,3-dichloropropanol) was a potent depletor of glutathione. 5. N-acetylcysteine was less efficient than glutathione as a nucleophile trap for epichlorohydrin, 1,3-dichloroacetone or reactive metabolites derived from 1,3-dichloropropan-2-ol. 6. 1,3-Dibromopropan-2-ol and 1,4 dibromobutan-2-ol were potent depletors of GSH but 1-bromopropan-2-ol produced less GSH depletion. Both dibromoalcohols depleted GSH when incubated with dialysed cytosol derived from the livers of untreated rats. 7. The GSH depletion mediated by 1,3-dichloropropan-2-ol, 1,3-dibromopropan-2-ol, 1,4-dibromobutan-2 ol and 1-bromopropan-2-ol was inhibited by inclusion of pyridine (1 mM) or cofactor omission. 1,3-Difluoropropanol did not deplete GSH under any of the conditions examined. PMID- 10379988 TI - Co-oxidation of acrylonitrile by soybean lipoxygenase and partially purified human lung lipoxygenase. AB - 1. Human lung lipoxygenase (HLLO) was partially purified by concanavalin-A (Con A) affinity chromatography that provided an easy and rapid one-step procedure for the removal (> or = 96%) of haemoglobin from cytosol. 2. HLLO exhibited dioxygenase activity towards arachidonic acid (AA) and linoleic acid (LA). The dioxygenase activity towards LA varied approximately 12-fold (48-591 nmol/min/mg protein) among different human lung samples examined. 3. Reverse-phase HPLC analysis of AA metabolites indicated the predominance of 15-lipoxygenase in human lung cytosol. 4. HLLO exhibited co-oxidase activity towards benzidine (BZD) and several other model compounds. The co-oxidase activity towards BZD was significantly inhibited by several lipoxygenase inhibitors. 5. HLLO and soybean lipoxygenase (SLO), used as a model enzyme, metabolized acrylonitrile (ACN) to 2 cyanoethylene oxide (CEO) and ultimately to cyanide. 6. HLLO was a approximately 6-fold better catalyst than SLO in converting ACN to cyanide. The generation of cyanide by HLLO was dependent on the concentration of enzyme and the reaction was inhibited by the lipoxygenase inhibitor nordihydroguaiaretic acid (NDGA) and the anti-oxidant butylated hydroxytoluene (BHT). 7. Under optimal assay conditions, the covalent binding of HLLO-generated reactive intermediate(s) from [14C]ACN to protein and DNA (nmol equivalent bound/15 min/mg HLLO/mg bovine serum albumin or calf thymus DNA) was observed at approximately 1.20+/-0.13 and 2.20+/-0.50 respectively. Both protein and DNA binding were inhibited by NDGA, butylated hydroxyanisole (BHA) and BHT. PMID- 10379990 TI - Preparation and properties of inhomogeneous hydroxyapatite ceramics. AB - Inhomogeneous ceramics of hydroxyapatite (HA) were prepared by sintering briquettes in which an inhomogeneous distribution of density was made by pressing HA powder into a die with rough walls. The resulting sample of such a ceramic is a hard thin shell with a loose core, and it is characterized by an inhomogeneous macro- and microstructure. It is a sintered conglomerate from HA grains containing grain boundary macropores in contact with the surface and micropores located inside the grains, part of which are also associated with the free surface. The highest value of microhardness is fixed on the surface of the sample. The radial distribution of microhardness in the (cylindrical) sample has an axisymmetric, nonmonotonic character and, on the whole, shows the decrease of microhardness (the increase of porosity) from the surface to the center. The highest values of microhardness, crushing strength, and fracture strength are close to those known for ceramics of moderate strength. PMID- 10379991 TI - Interaction of the anticancer agent Taxol (paclitaxel) with phospholipid bilayers. AB - Taxol (paclitaxel), a promising agent for use in ovarian and breast cancer, was incorporated into lipid vesicles (liposomes) composed of different saturated and unsaturated phosphatidylcholines, as well as saturated phosphatidylcholines mixed with the anionic phospholipid 1,2-dimyristoyl-sn-glycero-3-phosphatidylserine (DMPS) at different molar ratios, to yield information about Taxol-liposome interactions. For the physicochemical characterization of the thermodynamic, structural, and dynamic properties of these mixtures, differential scanning calorimetry (DSC), steady-state fluorescence depolarization, and Fourier transform IR spectroscopy was used. Time-dependent DSC measurements on 1,2 dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC)/Taxol mixtures of different concentrations were performed to yield information on the long-term stability of Taxol-liposome complexes. Partitioning of Taxol into saturated lipid bilayers results in changes of membrane physical properties, such as phase transition temperatures and lipid order parameter, that are different from those observed for unsaturated and charged phospholipid bilayers. Taxol incorporated into saturated phospholipids changes their thermotropic phase behavior: it reduces the lipid order parameter (i.e., has a "fluidizing" effect) in the gel phase of the lipid bilayers. On the contrary, partitioning of Taxol into unsaturated fluid phospholipid bilayers has a slight "rigidization" effect. The saturated lipid bilayer systems DPPC and 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine/DMPS have been identified with the highest incorporation efficiency for Taxol and are thus candidates for drug vehicles that can improve the therapeutic efficacy of Taxol. PMID- 10379992 TI - Characterization of cortical astrocytes on materials of differing surface chemistry. AB - The behavior of cortical astrocytes was evaluated on a number of medically relevant materials of differing physicochemical properties. This study describes cell attachment, DNA synthesis, production of extracellular matrix (ECM) proteins, and neuronal interactions of perinatal rat astrocytes in vitro. The number of attached astrocytes initially differed among the materials, decreasing with increasing material hydrophobicity. In contrast, the rate of DNA synthesis increased with increasing material hydrophobicity. With the exception of only one material, astrocytes reached confluence by 12 days in culture on all the materials tested. Furthermore, the expression of characteristic ECM proteins and the fundamental ability of astrocytes to support neuronal attachment and growth was qualitatively identical between populations of astrocytes on different materials. The ability of astrocytes to colonize different surfaces initially was mediated via adsorbed serum proteins, as reducing the capacity of a model surface to adsorb proteins inhibited astrocyte colonization for up to 2 weeks in culture. We propose that astrocytes are relatively insensitive to differences in surface chemistries so long as the proteins necessary for cellular attachment are capable of adsorbing to the material to some extent. It seems likely that the ability of astrocytes to produce and remodel a matrix creates a surface environment that eventually becomes similar regardless of the surface chemistry of the underlying material. PMID- 10379993 TI - Synthesis and characterization of dextran-maleic acid based hydrogel. AB - A new class of hydrogel precursor, dextran-maleic acid (Dex-MA), was synthesized by the reaction of dextran with maleic anhydride in the presence of the catalyst triethylamine. The effects of temperature, time, catalyst amount, and reactant concentration on the degree of substitution (DS) by MA was studied to establish an optimum reaction condition. The new hydrogel precursor had excellent solubility in various common organic solvents. The hydrogels based on Dex-MA precursor were made by the irradiation of Dex-MA with a long wave UV lamp. The Dex-MA hydrogels showed a very high swelling ratio in water, and the magnitude of swelling depended on the pH of the medium and the DS by MA. The Dex-MA hydrogels exhibited the highest swelling ratio in neutral pH, followed by acidic (pH 3) and alkaline pH (10). The most distinctive characteristic of Dex-MA hydrogels was that a carboxylic acid group was generated by the reaction of dextran with maleic anhydride. As a result, the swelling ratio increased with an increase of the DS of the MA segment (ionizable moiety that affects swelling ratio) in the Dex-MA hydrogel. PMID- 10379994 TI - Forced adhesive growth of K562 leukemic cells that normally grow in suspension induces variations in membrane lipids and energy metabolism: a proton NMR study. AB - The mechanisms responsible for the adhesion of cells onto a material's surface and the effects that that adhesion may have on cell structure and function are fundamental questions in biomaterials research. We recently demonstrated that the erythroleukemic cell line K562, which normally grows in suspension, can be induced to grow attached to a polylysine-coated solid surface in an anchorage dependent manner. In this study, the effects of the growth of K562 cells onto polylysine were further investigated utilizing 500 MHz 1H-NMR spectroscopy. The NMR results showed that when K562 cells are grown attached to a positively charged polylysine surface, there are alterations in lipids and energy metabolism. In particular, there was a 31% increase in phosphatidylcholine and a 15% decrease in each of its two precursors, glycerophosphatidylcholine and choline, as well as a 20% increase in CH2 lipids and a 7% decrease in CH3 lipids in treated cells compared to the controls. These results suggest that adhesive growth can induce strong variations in membrane structure, including the membrane fluidity of K562 cells. In addition, in cells attached to polylysine there was about a 10% decrease in creatine (together with phosphocreatine), a 20% increase in gamma-glutamate, a 15% increase in beta-glutamate, and a 24% decrease in lactate. This second set of results, which is closely related to energy metabolism, indicates that not only does adhesive growth induce changes in K562 cell membrane structure, but also in the utilization of energy in these cells. The data are discussed in view of the possible role played by surface charge in affecting cell structure and function in cells that come into direct contact with charged biopolymers. PMID- 10379995 TI - Optimal conditions for alumina coating formation on the MA956 superalloy for prosthetic bearing applications. AB - An experimental study of the oxidation treatment at high temperature of the ODS MA956 superalloy was conducted in an attempt to achieve a protective alumina scale for biomedical applications. A quadratic response-surface model was developed in order to study the effects of treatment time and temperature (in the range of 1000 degrees C to 1250 degrees C) on scale thickness. The obtained model adequately represents the experimental response and shows that the thickness gradients of the layer increase with the temperature for each exposure time and decrease steadily to zero as the treatment time increases. The microstructural characterization reveals that the alumina scale formed at or above 1000 degrees C consists of an alpha-alumina phase. Treatments at temperatures above 1150 degrees C give rise to an alumina scale with some defect probability. An increase in the temperature up to 1200 degrees C gives rise to the appearance of some blistering of the superficial scale. An oxidation treatment of 100 h at 1100 degrees C was found to be the best for guaranteeing the formation of a defect-free, compact, adherent, and continuous alpha-alumina scale thick enough to support satisfactory wear and biological conditions. PMID- 10379996 TI - The role of plasma proteins and stress in the assessment of hemocompatibility. AB - The physiological and psychological conditions of subjects supplying blood for hemocompatibility tests significantly affect the behavior of platelets in terms of both adhesion and activation. The responses of platelets to a standard biomaterial, polyethylene (PE), were examined with blood collected from male rabbits both in basal conditions and after stress. Different media were utilized. First, platelet-rich plasma (PRP) was used to obtain a PE response to contact with platelets. Then platelets drawn from PRP were isolated and washed with Krebs Ringer solution. One aliquot was suspended in serum (Pw-S) where fibrinogen was absent, another aliquot in Krebs-Ringer solution (Pw-KR) (in order to avoid the influence of the plasma proteins on platelets), and a third aliquot in the original plasma from which the platelets were drawn (Pw-PPP) (in order to restore the initial condition of the plasma but with washed platelets). The analysis of platelet adhesion and morphology was performed by Scanning Electron Microscopy (SEM). Differences in platelet adhesion and morphology were observed with four different media in nonstressed animals, with Pw-PPP showing a higher number and Pw-S and PW-KR lower numbers. Platelet morphology indicated low levels of activation. The platelets drawn from stressed subjects could not be counted in either PRP or PPP medium because they were fully aggregated and adhered; in contrast, in Pw-KR and Pw-S, no significant differences were found with respect to nonstressed conditions, and there was little difference in platelet morphology. All of these factors underline the role of plasma proteins, in particular fibrinogen, in the stress-induced activation of platelets. PMID- 10379997 TI - Characterization of rhBMP-2 pharmacokinetics implanted with biomaterial carriers in the rat ectopic model. AB - Recombinant human bone morphogenetic protein-2 (rhBMP-2) is a member of the bone morphogenetic protein family involved in de novo bone induction. Successful use of rhBMP-2 requires implantation with a biomaterial which can act as a scaffold for cell invasion for osteoinduction and retains rhBMP-2 at a site of implantation. This study was carried out to characterize rhBMP-2 pharmacokinetics from a variety of biomaterial carriers in a rat ectopic model. Retention of rhBMP 2 within carriers after 3 h was variable among the carriers (range, 75-10%), with collagenous sponges retaining the highest fraction of implanted dose. A gradual loss of rhBMP-2 was subsequently observed, the kinetics of which was strongly dependent on the implanted carrier. Collagenous carriers were observed to lose rhBMP-2 gradually from the implant site, whereas some of the mineral-based carriers retained a fraction of implanted rhBMP-2 within the implants. These differences in protein pharmacokinetics among carriers, in addition to their physicochemical nature, are expected to affect the biological activity of implanted rhBMP-2. PMID- 10379998 TI - Use of small intestinal submucosal implants for regeneration of large fascial defects: an experimental study in dogs. AB - A single layer of porcine small intestinal submucosa (SIS) was sutured into a 5 x 5 cm window created in the fascia lata of ten adult mongrel dogs in order to determine the efficacy of this material in promoting tissue regeneration of large fascial defects. A similar defect in the contralateral limb was left empty and served as a negative control. Tissue regeneration was examined grossly and histologically at 6 and 12 weeks. By 6 weeks, marked fibroplasia and angiogenesis had occurred throughout the SIS scaffold. The regenerated tissue was well organized and showed good integration with the adjacent fascia while the control specimens were filled with loose areolar connective tissue. At 12 weeks the experimental defects were filled with a regenerated tissue that grossly and histologically resembled normal fascia. There was no evidence of adhesions to the underlying musculature. Conversely, the tissue that filled the control defects remained disorganized and was markedly thinner than the adjacent fascia. The results of this study suggest that SIS is capable of supporting tissue regeneration in large fascial defects. The ability of this material to induce regeneration of a substantial area of tissue grossly and histologically similar to normal fascia and without adhesions to the underlying musculature makes its application in reconstructive surgery appear promising. PMID- 10379999 TI - Attachment of fibroblasts on smooth and microgrooved polystyrene. AB - In this study rat dermal fibroblasts (RDFs) were cultured on smooth or microgrooved (1-20 microm wide, 0.5-5.4 microm deep) substrates. Polystyrene microgrooved substrates were produced by solvent casting on molds that had been produced by photolithographic techniques. We investigated the attachment of RDFs with various analytical techniques. Light microscopy and image analysis showed that RDFs were oriented on most microgrooves. The rate of orientation effectively was increased by an increase of groove depth. An analysis of confluent layers of RDF showed that at confluency microgrooves were able to support greater numbers of cells. However, the largest numbers of cells were not found on the narrowest and deepest microgrooves even though such microgrooves have the largest total surface and induce the strongest alignment. Interference reflection microscopy (IRM) showed that the RDFs form focal adhesions where the cell membrane is only 10 nm from the substrate. IRM also showed that RDFs follow the contours of shallow and wide microgrooves but bridge the grooves on deeper and narrower ones. This could explain why such grooves are not able to increase the numerical cell adhesion to a greater degree. The absence of contact between cells and the bottom of the grooves is a very important factor in establishing contact guidance. PMID- 10380000 TI - Silica coatings formed on noble dental casting alloy by the sol-gel dipping process. AB - The sol-gel dipping process, in which liquid silicon alkoxide is transformed into the solid silicon-oxygen network, can produce a thin film coating of silica (SiO2). The features of this method are high homogeneity and purity of the thin SiO2 film and a low sinter temperature, which are important in preparation of coating films that can protect from metallic ion release from the metal substrate and prevent attachment of dental plaque. We evaluated the surface characteristics of the dental casting silver-palladium-copper-gold (Ag-Pd-Cu-Au) alloy coated with a thin SiO2 film by the sol-gel dipping process. The SiO2 film bonded strongly (over 40 MPa) to Ti-implanted Ag-Pd-Cu-Au alloy substrate as demonstrated by a pull test. Hydrophobilization of Ti-implanted/SiO2-coated surfaces resulted in a significant increase of the contact angle of water (80.5 degrees) compared with that of the noncoated alloy specimens (59.3 degrees). Ti implanted/SiO2-coated specimens showed the release of many fewer metallic ions (192 ppb/cm2) from the substrate than did noncoated specimens (2,089 ppb/cm2). The formation of a thin SiO2 film by the sol-gel dipping process on the surface of Ti-implanted Ag-Pd-Cu-Au alloy after casting clinically may be useful for minimizing the possibilities of the accumulation of dental plaque and metal allergies caused by intraoral metal restorations. PMID- 10380001 TI - Composition and structure of the apatite formed on PET substrates in SBF modified with various ionic activity products. AB - An apatite layer was formed on polyethyleneterephthalate (PET) substrates by the following biomimetic process. PET substrates were placed on granular particles of a CaO-SiO2-based glass in simulated body fluid (SBF) with ion concentrations nearly equal to those of human blood plasma to form apatite nuclei on their surfaces (first treatment). They then were soaked in modified SBFs, the ion concentrations of which were changed to give a variation in ionic activity product of apatite (IP), in order to make the apatite nuclei grow (second treatment). The Ca/P atomic ratio and the lattice constant c of the formed apatite decreased from 1.54 to 1.40 and from 6.880 to 6.838 A, respectively, with increasing ion concentrations from 0.75 to 2.00 times those of SBF, that is, with increasing IP from 10(-96.6) to 10(-91.9). This was attributed to an increase in the concentration of HPO4(2-) ion substituting for the PO4(3-) ion sites, which gave an increase in the Ca2+ in the apatite. Even the apatite formed in 1.00 SBF showed a Ca/P ratio of 1.51 and lattice constants a of 9.432 A and c of 6.870 A. The Ca/P ratio and lattice constant c were smaller and the lattice constant a was larger than those of the bone apatite; its Ca/P ratio and its lattice constants a and c, were 1.65, 9.419 A, and 6.88 A, respectively. This was attributed to the lower content (2.64 wt%) of the CO3(2-) ion substituting for the PO4(3-) ion sites of the apatite compared to that of the bone apatite (5.80 wt%). The lower content of the CO3(2-) ion in the apatite might be caused by the lower concentration of HCO3- ion in 1.00 SBF compared to that in human blood plasma. PMID- 10380002 TI - In vitro degradation of thin poly(DL-lactic-co-glycolic acid) films. AB - This study was designed to investigate the in vitro degradation of thin poly(DL lactic-co-glycolic acid) (PLGA) films for applications in retinal pigment epithelium transplantation and guided tissue regeneration. PLGA films of copolymer ratios of 75:25 and 50:50 were manufactured with thickness levels of 10 microm (thin) and 100 microm (thick). Degradation of the films occurred during sample processing, and thin films with a higher surface area to volume ratio degraded faster. Sample weight loss, molecular weight loss, dimensional, and morphological changes were analyzed over a 10-week period of degradation in 0.2 M of phosphate-buffered saline (PBS), pH 7.4, at 37 degrees C. All PLGA films degraded by heterogeneous bulk degradation. Sample weights remained relatively constant for the first several weeks and then decreased dramatically. The molecular weights of PLGA films decreased immediately upon placement in PBS and continued to decrease throughout the time course. PLGA 50:50 films degraded faster than 75:25 films due to their higher content of hydrophilic glycolic units. The results also demonstrated that thick films degrade faster than corresponding thin films with the same composition. This was attributed to the greater extent of the autocatalytic effect, which further was confirmed by heterogeneous gel permeation chromatograms. These studies suggest that the degradation rate of thin films can be engineered by varying film thicknesses. PMID- 10380003 TI - Incorporation of bovine serum albumin in calcium phosphate coating on titanium. AB - Calcium phosphate (Ca-P) and bovine serum albumin (BSA) were coprecipitated as a coating on commercially pure titanium (cpTi) with a high protein loading (15 wt %) by employing a recently developed wet-chemistry technique. It was observed that the incorporation of BSA significantly modified the morphology, composition, and crystallinity of the Ca-P coating. The Ca-P coating without BSA is a mixture of hydroxyapatite (HA) and octacalcium phosphate (OCP) with sharp-edged thin OCP crystal plates on the top layer, whereas only an HA phase was detected in the Ca P/BSA coating. The crystal plates in the latter had a more rounded appearance. The Ca-P/BSA coatings were immersed respectively in neutral (pH 7.4) and acidic (starting pH 4.0) phosphate-buffered saline (PBS) at 37 degrees C over a 14-day period. No protein release was detected in the neutral PBS during the immersion; however, a continuous release of BSA was measured in the acidic PBS, subsequently leading to the formation of a very dense and well-adherent composite coating of BSA and Ca-P on cpTi. The present study provides the possibility to achieve a long-term effective release of biologically active proteins from a Ca-P-coated metallic implant. PMID- 10380004 TI - Bone defect healing enhanced by ultrasound stimulation: an in vitro tissue culture model. AB - Ultrasound has many medical applications. Previous animal and clinical studies have clearly shown a positive effect of ultrasound on the rate of osseous repair. The present in vitro study was designed to elucidate the specific response of bony tissue to ultrasound treatment. Bilateral femora were obtained from 36 mature male Wistar rats. A bone defect was created at the center of each distal metaphysis. The femora were maintained for either 7 or 14 days in in vitro tissue culture and received 15 min of ultrasound stimulation or a sham exposure. The ultrasound intensity used was either 320 or 770 mW/cm2. Healing of the bone defect was evaluated by histomorphological examination and by analysis for the synthesis and secretion of prostaglandin E2. The results showed that ultrasound stimulation can accelerate both defect healing and trabecular bone regeneration. All experimental femoral defects treated with ultrasound healed faster than the untreated cortical defects, but only the defects receiving 770 mW/cm2 reached a level that was significantly different. The healing rate for the 320-mW/cm2 stimulated defects was intermediate between that of the 770-mW/cm2 and sham exposed defects. With ultrasound stimulation, prostaglandin E2 secretion by the experimental femora decreased significantly. Changes in the prostaglandin synthesis and concentration were found to correspond to changes in the amount of trabecular regeneration and to acceleration of bone healing. This highly controlled and well-studied model of ultrasound stimulation of bone healing in vitro can be used to further examine the biological mechanisms involved. PMID- 10380005 TI - Silicon nitride coating on titanium to enable titanium-ceramic bonding. AB - Failures that occur in titanium-ceramic restorations are of concern to clinicians. The formation of poorly adhering oxide on titanium at dental porcelain sintering temperatures causes adherence problems between titanium and porcelain, which is the main limiting factor in the fabrication of titanium ceramic restorations. To overcome this problem a 1-microm thick Si3N4 coating was applied to a titanium surface using a plasma-immersion implantation and deposition method. Such a coating serves as an oxygen diffusion barrier on titanium during the porcelain firings. The protective coating was characterized in the as-deposited condition and after thermal cycling. Cross sections of Ti/Si3N4-porcelain interface regions were examined by various electron microscopy methods and by energy dispersive analysis of X-rays to study the Si3N4 film's effectiveness in preventing titanium oxidation and in forming a bond with porcelain. The experiments have shown that this Si3N4 coating enables significant improvement in Ti-ceramic bonding. PMID- 10380006 TI - Crosslinked polyanhydrides for use in orthopedic applications: degradation behavior and mechanics. AB - High-strength, surface-eroding polymers were synthesized from methacrylated anhydride monomers of sebacic acid (MSA) and 1,6-bis(carboxyphenoxy) hexane (MCPH). These multifunctional monomers were photopolymerized using ultraviolet light to produce highly crosslinked polyanhydride networks. Through this approach, the crosslinking density of the resulting polymer network was used to control the final mechanical properties, while the degradation time scale was controlled by the chemical composition of the network. The combined hydrophobicity of the polymer backbone with the hydrolytically labile anhydride linkages led to surface-eroding networks, as confirmed by linear cumulative mass loss profiles as a function of degradation time for crosslinked polymer disks. By copolymerizing varying amounts of MSA and MCPH, the degradation rate of the final network was controlled from 2 days to 1 year. The tensile modulus of crosslinked poly(MSA) (1.4 GPa) was nearly an order of magnitude larger than that of linear poly(sebacic acid). In general, the mechanical properties of the crosslinked polyanhydrides networks were within ranges of those reported for cortical and trabecular bone. However, unlike bulk degrading polyesters such as poly(lactic acid), these surface eroding networks maintained >70% of their tensile modulus with 50% mass degradation. PMID- 10380007 TI - In vivo biocompatibility and mechanical study of novel bone-bioactive materials for prosthetic implantation. AB - Two epoxy materials with or without adhesively bonded hydroxyapatite (HA) coatings were studied for their biocompatibility and mechanical pushout strength using in vivo implantation in the rabbit lower femur for a duration of 10 days to 6 months. Both were two-part epoxies cured at room temperature for 24 h, with material 1 (Ampreg 26; SP Systems Limited, Cowes, UK) postcured at 110 degrees C (Tg approximately 80 degrees C) and Material 2 (CG5052; Ciba Geigy Limited, Cambridge, UK) at 125 degrees C (Tg approximately 120 degrees C). Implantation in dead rabbit bone was performed to provide mechanical baseline levels. Polymethylmethacrylate (PMMA) and conventionally HA-coated titanium alloy (Ti-6Al 4V) were used as control materials. In the biological study, different fluorescent dyes were used to label newly formed bone. After 6 weeks of implantation, results from mechanical pushout tests showed that the interfacial shear strength (ISS) values were significantly higher than for dead bones with each of the different implants (p < .01-.001). HA-coated material 2 showed a significantly higher ISS value than the uncoated material (p < .05) after 6 weeks' implantation. However, the ISS value for the uncoated material 2 was significantly higher than for PMMA controls (p < .05). No significant differences in the ISS values were shown between HA-coated materials 1 and 2 and Ti-6Al-4V on in vivo implantation for 6 weeks. Failure points of the pushout test from the three HA-coated materials were defined by scanning electron microscopy. Specimens implanted with both HA-coated epoxies were fractured within the HA-coatings or the bone, while with HA-coated Ti-6Al-4V cracked between the coating and metal implant. The percentage of bone in contact with the implant surface was obtained by image analysis which showed that there were no significant differences between different materials after short time implantation (up to 6 week). Long-term implantation of the HA-coated material 2 showed that the percentage of bone contact had increased from 52.8+/-1.1% (6 week) to 80.0+/-0.3% (3 months) (p < .01) and remained at 81.0+/-0.8% (6 months). Measurements of bone mineralization rate (BMR) showed that after 3 weeks of implantation, there were no significant differences between PMMA and uncoated materials 1 and 2. After 6 weeks, the BMRs in animals implanted with either HA-coated material 1 or 2 were significantly higher than with HA-coated Ti-6Al-4V (p < .05-.0001 in both cases), but with HA coated material 2 was lower than with this material uncoated (p < .05-.001). No significant differences were found between the two HA-coated epoxy materials. In addition, there were always lower BMRs during the third week of implantation than other periods regardless of biomaterial implanted. The study indicated that the adhesively bonded HA-coated novel epoxy materials were superior to conventional plasma-sprayed Ti-6Al-4V implants with respect to both BMR and bone integration with the implant surfaces. Adhesively bonded HA-coated epoxy materials had similar ISS values to HA-coated Ti-6Al-4V, but the former failed within the bone and coating, while the latter showed splitting between coating and metal. PMID- 10380008 TI - Bone-bonding behavior of alumina bead composite. AB - Previously we developed an alumina bead composite (ABC) consisting of alumina bead powder (AL-P) and bisphenol-alpha-glycidyl methacrylate (Bis-GMA)-based resin and reported its excellent osteoconductivity in rat tibiae. In the present study, are evaluated histologically and mechanically the effect of alumina crystallinity on the osteoconductivity and bone-bonding strength of the composite. AL-P was manufactured by fusing crushed alpha-alumina powder and quenching it. The AL-P was composed mainly of amorphous and delta-crystal phases of alumina. Its average particle size was 3.5 microm, and it took a spherical form. Another composite (alpha ALC), filled with pure alpha-alumina powder (alpha AL-P), was used as a referential material. The proportion of powder added to each composite was 70% w/w. Mechanical testing of ABC and alpha ALC indicated that they would be strong enough for use under weight-bearing conditions. The affinity indices for ABC, determined using male Wistar rat tibiae, were significantly higher than those for alpha ALC (p < 0.0001) up to 8 weeks. Composite plates (15 x 10 x 2 mm) that had an uncured surface layer on one side were made in situ in a rectangular mold. One of the plates was implanted into the proximal metaphysis of the tibia of a male Japanese white rabbit, and the failure load was measured by a detaching test 10 weeks after implantation. The failure loads for ABC on its uncured surface [1.91+/-1.23 kgf (n = 8)] were significantly higher than those for alpha ALC on its uncured surface [0.35+/-0.33 kgf (n = 8); (p < 0.0001)], and they also were significantly higher than those for ABC on the other (cured surface) side (p < 0.0001). Histological examinations using rabbit tibiae revealed bone ingrowth into the composite only on the uncured surface of ABC. This study revealed that the amorphous phase of alumina and formation of an uncured surface layer are needed for the osteoconductive and bone-bonding ability of ABC. ABC shows promise as a basis for the development of a highly osteoconductive and mechanically strong biomaterial. PMID- 10380009 TI - Effect of bioactive glass particle size on osseous regeneration. PMID- 10380010 TI - Independently ligating CD38 and Fc gammaRIIB relays a dominant negative signal to B cells. AB - CD38 is expressed on a variety of hematopoietic cells and has a unique enzymatic activity that converts nicotinamide adenine dinucleotide (NAD) into cyclic ADP ribose (cADPR) and then into ADPR. CD38 is expressed at increasingly higher levels on B cells at each stage of B cell differentiation, and is then down regulated on germinal center B cells and mature plasma cells. Crosslinking of CD38 on the surface of mature, resting B cells induces B-cell proliferation, which is enhanced by co-signals such as IL-4 and LPS. CD38-induced proliferation is abrogated by Fc gammaRIIB ligation and this inhibition can be effected by the addition of anti-Fc gammaRII Ab midway through a 48 h in vitro culture indicating that it delivers a potent negative signal to CD38 activated B cells. The suppressive signal was shown to occur through the Fc gammaRIIB because CD38 induced B-cell activation was not inhibited by the ligation of Fc gammaRIIB in Fc gammaRII-deficient B cells. These results indicate that Fc gammaRIIB can act as a regulatory molecule that modulates CD38 signals in vivo. PMID- 10380011 TI - Human immune response to a peptide mimic of Neisseria meningitidis serogroup C in hu-PBMC-SCID mice. AB - An anti-idiotype-based peptide mimic vaccine for Neisseria meningitidis serogroup C polysaccharide (MCPS) has been developed and shown to induce a response in mice that is specific, functional, and T-dependent. In this study, the immunogenicity of the MCPS peptide mimic vaccine preparation, as a potential vaccine for use in humans, is shown using the hu-PBMC-SCID mouse model. The human antibody response to the MCPS peptide mimic vaccine is specific and functional as shown by inhibition enzyme-linked immunoadsorbent assay (ELISA) and bactericidal assay. These data support the usefulness of the peptide mimic vaccine strategy for humans. PMID- 10380013 TI - Anti-P30-52 monoclonal antibody cross-reacted to Env V3 and inhibited the viral multiplication of HIV-1-infected MT-4 cells. AB - It is well known that the anti-p17 antibody titer decreases with the disease progression among human immunodeficiency virus type 1 (HIV-1) carriers. We previously established several murine anti-p17 monoclonal antibodies (MAbs) to investigate the immunological role of p17, and to further characterize these MAbs, we examined the anti-p17 antibody titer in serum of a patient who was a long-term nonprogressor with hemophilia, and found that the antibody for the p17 derivative peptide from amino acid residues 30 to 52 (P30-52) cross-reacted to the third variable region of the envelope glycoprotein of HIV-1, Env V3. In the present study, we primed mice with P30-52 to establish anti-P30-52 MAbs (P30-52 MAbs), and examined their affinity and whether they suppressed the viral multiplication of HIV-1-infected MT-4 (HTLV-1-transformed CD4+ T-cell line) cells, in a TCID50 assay. At the same time, an anti-Env V3 MAb (Env V3 MAb) was also established and examined as above. The IgM-type P30-52 MAb and Env V3 MAb showed heteroclitic binding, and the IgM-type P30-52 MAb inhibited the viral multiplication. We also found that an increase of fragmented DNA of HIV-1 infected MT-4 cells co-cultured with P30-52 MAbs. Because DNA fragmentation is one of the features of programmed cell death, the viral multiplication may be suppressed by the apoptosis of HIV-1-infected MT-4 cells co-cultured with P30-52 MAbs. Though the relationship between cross-reactivity and the inhibition mechanism of multiplication of HIV-1 is unclear, P30-52 of p17 may well be a useful region of viral proteins for the development of therapeutic and vaccination strategies. PMID- 10380014 TI - Cross-reactivity of anti-HIV-1-p17-derivative peptide (P30-52) antibody to Env V3 peptide. AB - Strong antibody responses are often seen in human immunodeficiency virus type 1 (HIV-1) carriers, but it is not known whether these antibodies are effective in the inhibition of disease progression. In this study, we examined antigenic epitopes for anti-HIV-1 p17 antibody (p17 Ab) in an HIV-1 carrier's serum, and found that the residues of amino acid numbers 1 to 12 (P1-12), 12 to 29 (P12-29) and 30 to 52 (P30-52) of p17 were highly recognized in the serum. Our examination of purified antibodies from the patient using the p17-derivative-peptide immunoaffinity columns showed that the reactivity of anti-p30-52 Ab (p30-52Ab) was high for p30-52 and the naive protein, p17. In addition, this P30-52Ab cross reacted with the third variable region of the envelope glycoprotein (Env V3). To confirm this cross-reactivity, we immunized mice with P30-52, and established a monoclonal antibody (MAb), 8H10. We found that 8H10 was also reactive to Env V3. It is unclear whether this cross-reactivity of P30-52 Ab can function as the inhibitor of HIV-1, but these results will be of help in clarifying the interaction of Env protein with HIV-1 gag polyprotein and the relationship of the decline of the p17 antibody titer with the disease progression in HIV-1 carriers. PMID- 10380012 TI - cDNA sequence analysis of monoclonal antibody FU-MK-1 specific for a transmembrane carcinoma-associated antigen, and construction of a mouse/human chimeric antibody. AB - Mouse monoclonal antibody (MAb) FU-MK-1, raised against a human gastric adenocarcinoma, recognizes a transmembrane antigen, GA733-2, present on most adenocarcinomas and seems to be of potential utility for immunodiagnosis and immunotherapy of those cancers. However, an inherent problem in their in vivo application is the human anti-mouse antibody response. In this study, we cloned and sequenced the variable region genes of the heavy and light chains (V(H) and Vkappa) of FU-MK-1 using the reverse transcription-polymerase chain reaction method. Then, we constructed a mouse/human chimeric antibody, designated as Ch FU MK-1, by fusing the FU-MK-1 V(H) and Vkappa genes to the human Cgamma1 and Ckappa genes, respectively, and by ligating the chimeric H and L chain genes to each other in a mammalian cell expression vector. The final gene construct was transfected into mouse non-Ig-producing hybridoma cells by electroporation. The Ch FU-MK-1 antibody thus prepared bound to human adenocarcinoma cells and competitively inhibited the binding of the parental FU-MK-1 to the adenocarcinoma cells. Ch FU-MK-1 also showed a potent antibody-dependent cell-mediated cytotoxicity (ADCC) with human peripheral blood mononuclear cells as effectors against the adenocarcinoma cells, indicating that this chimeric antibody seems to be suitable for in vivo therapeutic approaches. PMID- 10380015 TI - Characterization of two monoclonal antibodies against porcine VCAM-1. AB - Immunization of mice with TNF alpha-activated porcine endothelial cells led to the characterization of two monoclonal antibodies (MAbs), 5F3 and 8A7, specific for porcine VCAM-1. Upon flow cytometry, both antibodies increasingly labeled endothelial cells according to their degree of activation. They bound a band of MW 80 kDa on Western blots of endothelial cells, which is the apparent molecular weight of porcine VCAM-1. It was determined by surface plasmon resonance that the antibodies are directed to different antigenic sites. It was also found that 5F3 competes for binding the antigen with a MAb previously characterized as binding domain 1 of porcine VCAM-1. Subsequently, 5F3, but not 8A7, was found to inhibit the adhesion of human B lymphocyte Ramos cells to porcine endothelial cells in vitro. These antibodies, which do not cross-react with human VCAM-1, might be useful for diagnostic or therapeutic purposes in xenotransplantation. PMID- 10380016 TI - Monoclonal antibodies directed against AFAP-110 recognize species-specific and conserved epitopes. AB - The actin filament-associated protein, AFAP-110, is a Src SH2/SH3 binding partner that can modulate changes in actin filament structure. AFAP-110 contains a carboxy terminal motif that facilitates actin filament interactions, as well as amino terminal protein binding motifs, including an SH3 binding motif, two SH2 binding motifs, and two Pleckstrin homology domains. Two monoclonal antibodies (MAbs) were developed that recognized epitopes in either the amino terminus (MAb 4C3) or the carboxy terminus (anti-AFAP-110) of AFAP-110. Site-directed mutations that change key proline residues to alanine in the SH3 binding motif and an adjacent proline-rich motif abrogated MAb 4C3 binding. These same mutations have been shown to prevent SH3 interactions between AFAP-110 and Src527F. These data indicate that MAb 4C3 recognizes an epitope that is part of the SH3 binding motif. Interestingly, MAb 4C3 is not efficiently reactive with mammalian homologs of AFAP-110. Sequence analysis of a putative cDNA clone that encodes the amino terminus of the human AFAP-110 isoform predicted a one amino acid difference within this epitope, indicating a mechanism for species-specific binding by MAb 4C3. A second, MAb anti-AFAP-110, recognizes AFAP-110 across species and binds to an epitope within the carboxy terminus. This epitope includes the 5th heptad repeat of the carboxy terminal, leucine zipper motif (amino acids 592-598)--a motif that facilitates self-associations and may regulate the function of AFAP 110. These MAbs will be useful for analyzing the effects of AFAP-110 upon cell morphology and actin filament integrity. In addition, the avian-specific MAb 4C3 may be useful for studying the effects of avian AFAP-110 constructs expressed in mammalian cells, by providing an internal epitope tag. PMID- 10380017 TI - Detection of phenotypic heterogeneity within the murine splenic vasculature using rat monoclonal antibodies IBL-7/1 and IBL-7/22. AB - The homing of lymphocytes into various peripheral lymphoid organs involves a complex set of interactions between the circulating lymphoid cells and the local endothelium. While the initial binding and the adhesion processes of lymphocytes leading to their homing to the lymph nodes have thoroughly been studied, relatively little is known about the lymphoid-endothelial interactions taking place in the spleen. Our aim was to isolate rat monoclonal antibodies (MAbs) against the endothelial cells of the mouse spleen. Using splenic stroma derived from irradiated mice as antigen, two new rat MAbs were isolated. The MAb designated as IBL-7/1 bound to the sinus-lining (littoral) cells in the red pulp, marginal zone, and to the T- and B-cell compartments of the white pulp, respectively. However, it did not react with the central arteriole in the periarteriolar lymphoid sheath (PALS). In contrast to this pattern, the IBL-7/22 MAb recognized a shared antigen expressed by the sinusoidal and arterial endothelium. In addition to the endothelial reactivity, the IBL-7/22 MAb also stained the reticular components of the PALS and red pulp, but not that of the follicles. In vivo labelling with fluorescein (FITC)-conjugated IBL-7/1 MAb followed by confocal microscopic analysis revealed that the antigen recognized was expressed on the luminal surface of the sinusoids. The treatment of mice with IBL-7/1 MAb did not result in the altered distribution of T and B cells. These two new MAbs may be valuable tools for the phenotypic analysis of splenic endothelium, and can be used for the identification of various endothelial cell subpopulations of the mouse spleen. PMID- 10380018 TI - Novel super-high affinity sheep monoclonal antibodies against CEA bind colon and lung adenocarcinoma. AB - We have developed technology to create monoclonal antibodies (MAbs) using lymphocytes from immunized sheep. The affinities of these sheep monoclonal antibodies (SMA) can be several orders of magnitude higher than mouse MAbs. This paper reports the development and validation of a modified enzyme-linked immunoadsorbent assay (ELISA) method to select high-affinity antibodies of the desired specificity at the first screen. Using this method, we have isolated high affinity SMA to carcinoembryonic antigen (CEA), a marker of colon cancer. Comparisons of our novel SMA with mouse MAbs using the new ELISA and BIAcore technology (BIAcore AB, Stevenage, Herts, UK) have confirmed that we have made super-high affinity antibodies to CEA. One of these has a t(1/2) for dissociation of 8 days, which could provide a longer therapeutic window than is available with murine monoclonals currently being used in the clinic. This antibody has a specific tissue staining profile; it thus appears to be an excellent candidate for use in the clinic. PMID- 10380020 TI - Production and characterization of a specific rubisco monoclonal antibody, and its use in rubisco quantification during Zantedeschia aethiopica spathe development. AB - Ribulose 1,5-bisphosphate carboxylase/oxygenase was purified from leaves of Zantedeschia aethiopica and used to immunize female Balb/c mice. Monoclonal antibodies (MAbs) were raised by hybridoma technology using Sp2/0 myeloma cells as fusion partner. A random selected IgG2a subclass MAb was purified from ascitic fluid by affinity chromatography on Protein A-Sepharose CL-4B, with a recovery of 84.3% and it was apparently homogeneous on native PAGE. The monoclonality of the purified MAb was determined by IEF. The MAb was highly specific for Rubisco from leaves of Z. aethiopica as determined by Western blotting and was used to determine the concentration of Rubisco protein by enzyme-linked immunoadsorbent assay (ELISA), at three distinct stages of Z. aethiopica spathe development and in the leaf. The results suggest de novo synthesis of Rubisco during the spathe regreening, which could explain, at least in part, the increase of photosynthetic activity observed during regreening. PMID- 10380019 TI - Construction and characterization of a chimeric fusion protein consisting of an anti-idiotype antibody mimicking a breast cancer-associated antigen and the cytokine GM-CSF. AB - Anti-idiotype antibody, 11D10 mimics biologically and antigenically a distinct and specific epitope of the high molecular weight human milk fat globule (HMFG), a cancer-associated antigen present in over 90% of breast tumor samples. To augment the immunogenicity of 11D10 without the aid of a carrier protein or adjuvant, we made a chimeric 11D10-GM-CSF fusion protein for use as a vaccine. An expression plasmid for 11D10 was made by ligation of the DNA sequences of the 11D10 light-chain variable region upstream of the human kappa constant region. The heavy-chain plasmid carrying GM-CSF was made by ligation of the heavy-chain variable region sequences upstream of the human gamma1 constant region CH1 fused to the DNA fragment encoding the mature GM-CSF peptide 3' to the CH3 exon. NS1 plasmacytoma cells were transfected with the light and heavy-chain vectors by electroporation. Fusion protein secreted in the culture medium was purified and was characterized by gel electrophoresis as well as by determination of the biological activity of the fused GM-CSF. In nonreducing SDS-polyacrylamide gels, a single band approximately 200 Kd reacted with anti-human kappa, anti-human lambda1 and anti-GM-CSF antibodies. In reducing polyacrylamide gels, a approximately 74 kd protein reacted with anti-human lambda1 and anti-GM-CSF antibodies. The fusion protein induced proliferation of GM-CSF dependent NFS-60 cells. These results suggest that the protein is a chimeric anti-idiotype antibody consisting of 11D10 variable domains, human kappa and lambda1 constant domains and that the GM-CSF moiety fused to the constant region lambda1 is biologically active. PMID- 10380021 TI - Intraoperative acetazolamide in the prevention of intraocular pressure rise after pars plana vitrectomy with fluid-gas exchange. AB - PURPOSE: To assess the effect of intraoperative acetazolamide (Diamox) on postoperative intraocular pressure (IOP) in gas-filled, vitrectomized eyes. METHODS: We conducted a prospective randomized clinical trial of 63 consecutive patients undergoing pars plana vitrectomy with total fluid-gas exchange and long acting intraocular gas tamponade. Patients were randomized by a blind draw to receive either intravenous 500 mg acetazolamide (Diamox) (Group 1) or no treatment (Group 2) at the conclusion of the operative procedure. Intraocular pressures at the conclusion of surgery (IOP-1), 4-8 hours following surgery (IOP 2), and on the first postoperative day (IOP-3) were measured using an Oculab Tono Pen. RESULTS: Patients in Groups 1 and 2 showed similar mean IOP on postoperative day 1 (20.48+/-7.84 mmHg versus 19.89+/-7.89 mmHg). A similar incidence of IOP-2 greater than 30 mmHg (1 versus 3 patients with high IOP) and IOP-3 greater than 30 mmHg (4 versus 3 patients with high IOP) was seen. Patients in Group 1 had a lower mean IOP at 4-8 hours postoperatively (16.25+/-6.47 mmHg) than those in Group 2 (20.13+/-6.33 mmHg). No correlation could be demonstrated between IOP-1 and subsequent IOP. However, IOP on the first postoperative day (IOP-3) was strongly correlated with IOP 4-8 hours after surgery (IOP-2) (P = 0.0001). No protective effect of Diamox could be demonstrated on either IOP-2 or IOP-3. CONCLUSIONS: No protective effect against pressure rise could be demonstrated for intraoperative acetazolamide (Diamox) in the prophylaxis of IOP rise following pars plana vitrectomy and total fluid-gas exchange with long-acting intraocular gas. PMID- 10380022 TI - Vitrectomy with silicone oil or long-acting gas in eyes with giant retinal tears: long-term follow-up of a randomized clinical trial. AB - PURPOSE: To determine if either silicone oil or perfluoropropane (C3F8) gas is superior to the other in the treatment of giant retinal tears complicated by proliferative vitreoretinopathy. METHODS: Forty-seven eyes with giant retinal tears were treated in a prospective randomized fashion. Twenty-two eyes were randomized to C3F8 gas and 25 eyes were randomized to silicone oil. Follow-up of 60 months is reported. Of the silicone oil-treated eyes, 9 had oil retained in the eye; oil was removed from the remaining 16 eyes. Successful anatomic attachment of the retina, final visual acuity, and surgical complications were the main outcome measures. RESULTS: There were no differences between the groups in any of the main outcome measures. CONCLUSIONS: This study showed that silicone oil and C3F8 gas are equal in most respects for the management of giant retinal tears with proliferative vitreoretinopathy. PMID- 10380023 TI - Early versus late removal of retained intraocular foreign bodies. AB - PURPOSE: To compare early versus late removal of retained intraocular foreign bodies (IOFBs). METHODS: Sixty-two patients presenting with open-globe injuries due to lacerations by retained IOFBs were consecutively operated by the same surgeon between 1989 and 1997. Minimum follow-up was more than 3 months. In 43 patients, the IOFB was removed during the first 24 hours after the accident. In 19 patients, in whom the wound had been closed during a first operation, the IOFB was removed later than 24 hours after the accident. The study groups did not vary significantly in age, refractive error, type and size of IOFB, prevalence of traumatic cataract and peripheral or central corneal lacerations, or visual acuity on presentation. RESULTS: Endophthalmitis developed significantly more often in the late intervention group than in the early intervention group (3/19 [15.7%] versus 1/43 [2.3%]; P = 0.0467; chi-square test). Considering patients with retinal lesions due to the IOFB (n = 47), proliferative vitreoretinopathy occurred significantly more often in the late intervention group than in the early intervention group (6/13 [46.2%] versus 6/34 [17.6%]; P = 0.0449). CONCLUSIONS: Confirming previous reports, the results of the current study suggest that removing retained IOFBs within the first 24 hours after the injury may in some clinical situations reduce the risks of infectious endophthalmitis and proliferative vitreoretinopathy. PMID- 10380024 TI - Visual field change in eyes with retinal pigment epithelial tear. AB - PURPOSE: To study the effects of retinal pigment epithelial (RPE) deprivation on retinal sensitivity with serial automated static perimetry in cases of RPE tear involving the foveal area. METHODS: Two eyes with a tear of the RPE were diagnosed as such on biomicroscopic and fluorescein angiographic examination. Static perimetry was performed in the follow-up study with the Humphrey field analyzer central 10-2 program. RESULTS: The first patient showed a dense scotoma corresponding to a defect in the RPE, which showed mild deterioration throughout the follow-up period from 2-11 weeks after the development of RPE tear. In contrast, the second patient showed preserved visual acuity and an absence of central visual field defects, despite an apparently denuded Bruch membrane involving the fovea during 8-month follow-up. CONCLUSION: Apparent RPE defect in eyes with RPE tears may or may not be associated with severe visual field defects. The pathophysiology of the disease should be studied, considering these perimetric findings. PMID- 10380025 TI - Ultrasound biomicroscopic features of anterior proliferative vitreoretinopathy. AB - PURPOSE: To evaluate the utility of ultrasound biomicroscopy (UBM) in the diagnosis of anterior proliferative vitreoretinopathy (APVR). METHODS: Ultrasound biomicroscopy (frequency, 50 MHz; depth of penetration, 5 mm; resolution, 40 microm) was used to examine 53 eyes of 52 subjects with rhegmatogenous or traumatic retinal detachment (RD). The results were compared with findings during vitrectomy. RESULTS: In 43 eyes (42 patients), RD was detected using UBM, attaining 81% consistency with observations during operation. Among 36 eyes with surgically proved APVR, 33 (92%) had APVR detected by UBM. The UBM images showed shallow RD, circumferential contraction, or anterior displacement around the ora serrata. The anterior displacement could be classified into C-shaped anterior displacement, ciliary body adhesion, and pupil adhesion. Ultrasound biomicroscopy showed that a retinal step was formed on the surface of the ora serrata and the ciliary body. Failure to detect APVR by UBM may be due to severe choroidal detachment or a narrow palpebral fissure that hampers exposure of the ora serrata and the part of the eyeball posterior to it. CONCLUSION: Ultrasound biomicroscopy is a useful tool for the evaluation of APVR, which is difficult by ordinary methods. Ultrasound biomicroscopic images of APVR have typical features that may be used to help guide surgery and estimate prognosis. PMID- 10380026 TI - Hypodermic needles: a new source of penetrating ocular trauma in Indian children. AB - PURPOSE: To study the clinico-microbiologic profile and visual prognosis of ocular injuries caused by disposable hypodermic needles used by children to squirt water. METHODS: We analyzed 19 consecutive cases of hypodermic needle injury seen at our institute. RESULTS: The average age of the patients was 10.3 years (range, 4-20 years). A small self-sealed corneal or scleral laceration was seen in 11 eyes; in 8 eyes, the site of injury was occult. Initial visual acuity was no light perception (3 eyes) or hand motion or light perception (16 eyes). Surgery in 18/19 eyes included vitrectomy with intraocular antibiotic injections for endophthalmitis (14 eyes), evisceration for panophthalmitis (2 eyes), and cataract extraction for traumatic cataract (2 eyes). Final visual acuity was no light perception or light perception only in 10 eyes, 20/400-20/60 in three eyes, and 20/40 or better in six eyes. CONCLUSIONS: Severe ocular morbidity may result from improper disposal of hypodermic needles. PMID- 10380027 TI - Penetration of topical and oral ciprofloxacin into the aqueous and vitreous humor in inflamed eyes. AB - PURPOSE: To assess the aqueous and vitreous penetration of ciprofloxacin after topical and combined topical and oral administration and investigate the effects of inflammation on drug penetration. METHODS: A standardized penetrating injury was made in the right eyes of 16 rabbits. Intraocular inflammation was induced by intravitreal injection of a suspension of Staphylococcus aureus in these eyes. The animals were divided into two groups according to treatment methodology: topical and topical-oral. The intact left eyes of the animals were maintained as controls. In the topical treatment group, two drops of ciprofloxacin 0.3% were instilled to both eyes every 30 minutes for 4 hours. In the topical-oral treatment group, animals were given two oral 40 mg/kg doses of ciprofloxacin at 12-hour intervals. After the last oral dose, the protocol of the topical group was applied to these eyes. Half an hour after the last drop, 100-microL samples were taken from aqueous and vitreous humor of all eyes. Drug concentrations were measured using high-pressure liquid chromatography. RESULTS: Mean aqueous levels of ciprofloxacin in control eyes were 2.31 microg/mL (range, 1.02-6.27 microg/mL) in the topical group and 5.88 microg/mL (1.52-17.81) in the topical-oral group. Mean aqueous levels in inflamed eyes were 7.36 microg/mL (2.34-17.15) in the topical group and 14.43 microg/mL (2.18-18.66) in the topical-oral group. Mean vitreous levels in control eyes were 0.77 microg/mL (0.09-1.93) in the topical group and 1.01 microg/mL (0.49-1.57) in the topical-oral group. Mean vitreous levels in inflamed eyes were 0.95 microg/mL (0.18-1.27) in the topical group and 1.98 microg/mL (0.51-3.34) in the topical-oral group. There was no significant difference among the groups (P > 0.05). Mean aqueous levels in all eyes and mean vitreous levels in the combined topical and oral group of inflamed eyes were above the 90% minimum inhibitory concentration for most of the common microorganisms causing endophthalmitis. CONCLUSION: There is an increase in both aqueous and vitreous humor concentrations with inflammation and with oral and topical administrations, as opposed to topical only, of ciprofloxacin. Using oral as well as topical treatment may be a beneficial method of antibiotic prophylaxis in ocular trauma once a patient has received intravenous or intravitreal therapy. PMID- 10380028 TI - Experimental prophylaxis of Staphylococcus aureus endophthalmitis after vitrectomy: the use of antibiotics in irrigating solution. AB - PURPOSE: To test the efficacy of clindamycin and gentamicin in irrigating solution during vitrectomy to prevent experimental Staphylococcus aureus endophthalmitis. MATERIALS AND METHODS: Thirty-six New Zealand white rabbits were divided into six groups. Vitrectomy using two different irrigating solutions was followed by intravitreal injection of S. aureus: Group 1, balanced salt solution (BSS) followed by 1,000 colony-forming units (CFU) S. aureus; Group 2, BSS fortified with clindamycin, 10 microg/mL, and gentamicin, 8 microg/mL (CGBSS), followed by intravitreal injection of 1,000 CFU S. aureus; Group 3, BSS followed by 2,000 CFU S. aureus; Group 4, CGBSS followed by 2,000 CFU S. aureus; Group 5, BSS followed by 4,000 CFU S. aureus; and Group 6, CGBSS followed by 4,000 CFU S. aureus. The eyes were examined clinically after surgery. Vitreous samples were cultured and histologic studies were performed. RESULTS: Severe endophthalmitis developed in all eyes in Groups 1, 3, and 5 (not given antibiotics). No endophthalmitis developed in Group 2. In Group 4, five of the six eyes were normal and one eye had endophthalmitis. In Group 6, one eye had clear vitreous and fundus, three eyes had moderate vitreous haze, and the other four eyes demonstrated severe endophthalmitis. Bacterial growth in Groups 1, 2, 3, 4, 5, and 6 were 4/4, 0/4, 6/6, 1/6, 4/6, and 2/8 eyes, respectively. CONCLUSION: When 1,000 to 2,000 CFU S. aureus were injected after vitrectomy, clindamycin and gentamicin in the irrigating solution significantly diminished the intraocular inflammation and the rate of positive bacterial culture. Clindamycin and gentamicin in the irrigating solution were not significantly effective when 4,000 CFU bacteria was injected; however, the degree of inflammation was less severe than in the control group. PMID- 10380029 TI - Tolerance of extended-term vitreous replacement with perfluoro-n-octane and perfluoroperhydrophenanthrene mixture (phenoctane). AB - PURPOSE: To improve the surface visibility of perfluoroperhydrophenanthrene, we added various percentages of perfluoro-n-octane. METHODS: Twenty New Zealand white rabbit eyes underwent gas vitrectomy. One milliliter of balanced salt solution was injected into each Group 1 eye as control. Perfluoro-n-octane was added to perfluoroperhydrophenanthrene in ratios of 15:85, 25:75, and 50:50; 1 mL of each mixture was injected into the vitreous cavities of Groups 2, 3, and 4, respectively. Eyes were examined clinically and electroretinograms were performed before and 4 and 8 weeks after injection, when the rabbits were killed. Eyes were processed for light and transmission electron microscopy (TEM). RESULTS: The indices of refraction of the 15:85, 25:75, and 50:50 mixtures were 1.3275, 1.3191, and 1.3026, respectively, and the mixtures were visible in the vitreous cavity. Emulsification and mild-to-moderate dust-like opacities were observed in eyes from Groups 2 through 4; vitreous strands formed in four of the Group 4 eyes. The retinae were attached. Electroretinographic responses were normal, except in one eye with cataract. Light microscopy showed normal retinal architecture, with some macrophages with intracytoplasmic vacuoles on the surface of the inferior retina or in the vitreous cavity. Fingerprint disfigurement of photoreceptor outer segments was seen in some Group 3 and 4 eyes under TEM. CONCLUSIONS: Minimal changes were induced by the 15:85 mixture in the rabbit eye. The mixtures of 25:75 and 50:50 produced some ultrastructural changes of the retina. The mixtures were visible under water. PMID- 10380030 TI - Current popularity of pneumatic retinopexy. AB - PURPOSE: To compare the popularity of pneumatic retinopexy (PR) in 1997 with its popularity in 1990 among retinal specialists. METHODS: In 1997, a survey was mailed to the 1994-1995 members of the Retina or Vitreous Societies who lived in the United States or Canada, asking how they would manage a hypothetical retinal detachment. The choices were limited to PR, segmental scleral buckling, scleral bucking with encircling, primary vitrectomy, and Lincoff balloon. The results of the survey were compared with those previously reported by a similar survey in 1990. RESULTS: The majority (55%) of respondents selected PR, which is a twofold increase over those who preferred it in 1990 (odds ratio 2.08; 95% confidence interval 1.53, 2.85). The popularity of PR was inversely proportional to the length of time the respondents had been in practice. If the eye with the hypothetical detachment had pseudophakia, only 30% of respondents selected PR. If the eye had additional tears, vitreous hemorrhage, or lattice degeneration, only about one-sixth preferred PR. CONCLUSION: Pneumatic retinopexy was much more popular in 1997 than it was in 1990. Its popularity continues to be influenced by the age of the surgeon and by the complexity of the detachment. PMID- 10380031 TI - Diagnostic and therapeutic challenges. Bilateral reduction in central visual function. PMID- 10380032 TI - Bilateral juxtafoveal telangiectasis in a family. PMID- 10380033 TI - Ocular and dermatologic findings in two siblings with mal de Meleda. PMID- 10380034 TI - Regression of retinal arterial aneurysms in a case of idiopathic retinal vasculitis, aneurysms, and neuroretinitis (IRVAN). PMID- 10380035 TI - In vivo formation of heavy oil. PMID- 10380036 TI - Free radical generation following pressure-induced retinal ischemia in the rat. PMID- 10380037 TI - Familial exudative vitreoretinopathy associated with monocular axial myopia. PMID- 10380038 TI - Intracranial hypertension associated with acquired hyperopia and choroidal folds. PMID- 10380039 TI - Retinal pigment epithelium lesions associated with choroidal ischemia in preeclampsia. PMID- 10380040 TI - Solitary fibrous tumor of the skin. AB - Solitary fibrous tumor (SFT) is an uncommon mesenchymal tumor that typically arises in the pleural cavity. Comprised of spindled cells characteristically arranged in diverse architectural patterns, SFT histologically simulates a variety of benign and malignant mesenchymal tumors. The diagnosis of SFT has been refined by the availability of newer immunohistochemical markers such as CD-34 and factor XIIIa, facilitating the identification of SFTs arising in multiple extrapleural sites, including the skin. We describe three cases of primary cutaneous SFT, review the literature, and discuss the histologic and immunohistochemical differential of other cutaneous tumors that SFT can mimic. PMID- 10380041 TI - Postoperative/posttraumatic spindle cell nodule of the skin: the dermal analogue of nodular fasciitis. AB - Spindle cell proliferations of the skin are diverse, both morphologically and mechanistically. The authors have encountered four examples of a distinctive reactive/reparative cutaneous spindle cell lesion that shows homology with ones seen in the genitourinary tract and oral cavity and that is known as "postoperative/posttraumatic spindle cell nodule" (PSCN). These occurred in the skin of the face and scalp (2 cases), arm (1 case), and vulvar skin (one case), and were clearly related historically to prior episodes of trauma. The proliferations were characterized by variably-apposed and cytologically-bland spindle cells with numerous mitotic figures, set in a highly vascular stroma containing extravasated erythrocytes and inflammatory cells. All lesions were immunoreactive for vimentin, actin, and desmin, with no labeling for keratin. Postoperative/posttraumatic spindle cell nodule of the skin is a significant pseudoneoplastic lesion that is related (and perhaps identical pathogenetically) to nodular fasciitis; as such, it may be mistaken for a sarcoma or a spindle cell carcinoma. Careful attention to clinicopathologic and histologic details should result in its accurate recognition. PMID- 10380043 TI - Epidermal changes in cutaneous lesions of sarcoidosis. AB - The essential histologic finding of cutaneous sarcoidosis is a noncaseating epithelioid cell granuloma in the dermis or, infrequently, in the subcutaneous tissues. However, there have been cases of cutaneous sarcoidosis with clinical appearances altered by epidermal changes such as ulcers, the formation of psoriasiform or verrucous plaques, and by hypopigmentation. In this study, histologic examinations of the epidermis were performed in cutaneous lesions of 62 cases of sarcoidosis. Seventy-nine percent (49/62) showed epidermal changes including hyperkeratosis (8/49), parakeratosis (10/49), acanthosis (6/49), and epidermal atrophy (35/49). Lymphoid cells extended into the epidermis in 50 of 62 cases. The infiltration patterns were in the forms of a spongiotic reaction (21/50), lichenoid tissue reaction (9/50), and simple exocytosis without epidermal vesiculation (20/50). Immunohistochemical studies showed that the lymphoid cells in the epidermis expressed CD3, 8, 45RO, and 11a. The epidermal changes overlying the granulomatous lesions contribute to the variety of clinical manifestations and are in part associated with the pathogenesis of cutaneous sarcoidosis. PMID- 10380042 TI - Amelanotic blue nevus: a variant of blue nevus. AB - Blue nevi are typically heavily melanized. We report a variant of blue nevus that is minimally pigmented. Of the 1,358 blue nevi seen in our laboratory during the last 6 years, 38 (2.7%) were selected that had scant or absent melanin. We refer to these blue nevi as the amelanotic type. Approximately half of the cases in clinical diagnosis were nevus of some type, whereas other differential diagnoses were basal cell carcinoma, dermatofibroma, and lesion. Histologically all specimens were characterized by the spindle-shaped cells seen in blue nevi, but with very little or no obvious melanin. Some lesions were markedly cellular, resembling the features of cellular blue nevus. No hemosiderin was identified on Perls' stain, whereas Fontana-Masson stain was variably positive. Usually there was fibrous stroma. In most cases, the histologic differential diagnosis was dermatofibroma. Other histologic differential diagnoses included amelanotic and/or spindle cell melanoma, dermal Spitz nevus, neurofibroma, and scar. There was no pleomorphism or increased mitotic activity. Evidence of epidermal melanocytic hyperplasia was seen in two cases. Furthermore, the lesions had been present for many years without evidence of recent change. Immunohistochemistry showed all cases to be strongly positive with anti Mel-5 antibody, but only weakly positive or negative with anti S-100 and HMB-45 antibodies. We would like dermatologists and pathologists to be aware of this unusual and uncommon entity. PMID- 10380044 TI - A clinical and histologic study of 37 cases of immunoglobulin A-associated vasculitis. AB - Immunoglobulin (Ig) A-associated vasculitis is commonly equated with the multiorgan systemic vasculitic syndrome Henoch-Schonlein purpura (HSP), which occurs predominantly in the pediatric age group. By natural language search of the databases of two outpatient dermatopathology practices, the authors selected for review 37 cases of IgA-associated vasculitis, 23 of which were associated with antecedent infection, most commonly of the upper respiratory tract. Criteria for a diagnosis of HSP were met in 15 cases, 13 of which were in the setting of prior infection. Lower extremity skin involvement was ubiquitous. A more widespread form of vasculitis was also seen, particularly in the setting of previous infection. Several of the patients with previous infection had underlying medical illnesses including rheumatoid arthritis, atopy, renal failure, lupus erythematosus, insulin dependent diabetes mellitus, autoimmune thyroid disease, and Wegener's granulomatosis. In those patients lacking an apparent microbial trigger, Sjogren's disease with anti-Ro antibodies and hypergammaglobulinemia, lupus erythematosus, inflammatory bowel disease, IgA paraproteinemia, bronchogenic and prostatic carcinoma, cryoglobulinemia, and lymphoma were uncovered. Regardless of whether an infectious stimulus was implicated, certain cofactors with the potential to enhance vascular injury were uncovered; these included anti-Ro antibodies, antineutrophil cytoplasmic antibody, diabetic microangiopathy, and a hyperviscosity state. In the infective group, a pustular vasculitis, defined as a neutrophilic vascular reaction in concert with epithelial pustulation, was seen in 81% of cases versus 33% in the noninfectious group (p = 0.02). The prototypic histomorphology in the noninfective group was one of a mild cell poor leukocytoclastic vasculitis; Vasculitis was of greater severity in patients with antecedent infection (p = 0.026). An infectious trigger, typically of mucosal origin, can frequently be identified in patients with cutaneous IgA-associated vasculitis, especially those with the symptom complex of HSP. The light microscopy appears to distinguish patients who have an infectious trigger from those who do not. IgA-associated vasculitis may be a clue to the presence of certain underlying disorders where there is immune dysregulation or enhanced susceptibility to immune complex entrapment. PMID- 10380045 TI - HIV-associated eosinophilic folliculitis in an infant. AB - HIV-associated eosinophilic folliculitis (HIV-EF), which is a well-known entity in adults, has not been described in children. Although Ofuji's disease (OD) or eosinophilic pustular folliculitis (EPF) has been described in children and shares histopathologic features with HIV-EF, it is a distinct entity with characteristic clinical features. We report the occurrence of eosinophilic folliculitis in an 8-month-old HIV-positive patient and discuss the clinical, pathologic and possible pathogenetic aspects thereof. In addition, differences in the clinical manifestations of the present case and that of I-EPF are addressed. Because of clinicopathologic similarities between the present case and HIV associated eosinophilic folliculitis (HIV-EF) in adults, we believe that eosinophilic folliculitis in this patient represents a cutaneous manifestation of HIV infection, rather than co-incidental occurrence of the infantile form of Ofuji's disease in an HIV-positive patient. PMID- 10380046 TI - Neutrophilic lobular (pustular) panniculitis associated with rheumatoid arthritis: a case report and review of the literature. AB - Rheumatoid nodules, which affect the subcutis around joints, are the most frequent specific cutaneous lesions of rheumatoid arthritis (RA). Panniculitis is a rarely reported and nonspecific complication of RA. We report a 42-year-old woman with seropositive RA who presented with a 2-month history of lower leg panniculitis. Biopsy of a leg nodule showed a lobular neutrophilic infiltrate with lipophages and central basophilic necrosis. In addition, focal changes of lipomembranous fat necrosis indicative of ischemic damage were identified at the margins of the lobular infiltrate. Neutrophilic lobular panniculitis is commonly detected in panniculitis secondary to bacterial infections, pancreatitis, and factitial causes. However, this pattern of panniculitis has also been reported in some cases of erythema nodosum-like lesions found in Behcet disease or bowel bypass syndrome and in rare cases of seropositive RA. These reported histologic findings fall into the spectrum of neutrophilic vascular reactions described by Jorizzo and Daniels for RA-associated dermatoses. In view of these findings. RA and related neutrophilic dermatoses (e.g., Behcet disease) should be included in the differential diagnosis of neutrophilic lobular panniculitis. PMID- 10380047 TI - Neurofollicular hamartoma with strong diffuse S-100 positivity: a case report. AB - Neurofollicular hamartoma is a recently described lesion with a distinct pilosebaceous and spindle cell proliferation. Neurofollicular hamartoma is composed of spindle cells haphazardly arranged in a fibromyxoid stroma closely associated with an abnormal hyperplasia of folliculosebaceous units. Although this histologic pattern has been classified as "neurofollicular," all cases reported thus far have had only scattered spindle cells with S-100 positivity. We present a case of neurofollicular hamartoma with strong and diffusely positive staining of spindled cells for S-100 protein. This lesion also shows scattered positivity of spindle cells for monoclonal neuron specific enolase and synaptophysin. We interpret the results of immunostains of this lesion as evidence for neural differentiation. This case validates the concept of "neurofollicular" hamartoma. PMID- 10380048 TI - Desmoplastic fibroblastoma (collagenous fibroma). AB - Desmoplastic fibroblastoma is a vary rare subcutaneous proliferation. We describe a case of desmoplastic fibroblastoma in a 24-year-old Korean woman who presented with a 2.5 cm solitary and firm nodule on her back which had been present for 3 months. Histologic studies showed a well demarcated subcutaneous tumor composed of stellate or spindle shaped cells embedded in hypovascular fibrous or fibromyxoid stroma. No mitotic figures, calcification, or necrosis were observed. The stellate or spindle shaped cells were positive for vimentin. The stroma stained positively with Alcian blue and Masson trichrome. S-100 protein, actin, desmin, and elastic fiber stains were all negative in the stellate or spindle shaped cells. There has been no recurrence or metastasis of the tumor over an 18 month follow-up. PMID- 10380050 TI - EORTC classification for primary cutaneous lymphomas: the best guide to good clinical management. European Organization for Research and Treatment of Cancer. AB - In 1997 the Cutaneous Lymphoma Study Group of the European Organization for Research and Treatment of Cancer (EORTC) published a proposal for a classification for the group of primary cutaneous lymphomas (1). The EORTC classification is the first and only classification that is designed exclusively for the group of primary cutaneous lymphomas. It is also the only classification that has been clinically validated for this group of diseases. As illustrated by this special issue, this classification has resulted not only in the discussion of the definition and terminology of some types of cutaneous T-cell lymphoma (CTCL) and cutaneous B-cell lymphoma (CBCL), but also in a discussion of whether organ-based classification schemes (separate from existing hematopathologic classification schemes for non-Hodgkin lymphomas) should be used. This article explains why it was necessary to create a separate classification for the group of primary cutaneous lymphomas. A short introduction on the history of the classification of cutaneous lymphomas is provided. Next, the basic principles of the EORTC classification are presented. Finally, controversies between the EORTC classification versus the REAL classification (2) and the proposed WHO classification (3), which still impede the usage of one common classification system, are discussed. PMID- 10380049 TI - Sweat gland proliferations in scleromyxedema. AB - Eccrine sweat duct proliferations may be found in various inflammatory and neoplastic skin lesions. We report a patient with scleromyxedema with extensive proliferations of intradermal sweat ducts. Three-dimensional reconstruction demonstrated extensive coiling and branching of the sweat ducts leading into cystic lacunae. In contrast to the basal cell carcinoma that had grown within the scleromyxedematous skin, the ducts close to the lumen stained positive for carcinoembryonic antigen and could therefore be differentiated from basal cell carcinoma. In micrographically controlled surgery of cutaneous epithelial tumors that are located in chronically inflamed skin, such sweat gland proliferations have to be considered as differential diagnosis. PMID- 10380051 TI - The revised European-American Classification of Lymphoid Neoplasms (REAL): a preferred approach for the classification of cutaneous lymphomas. AB - The Revised European-American Classification of Lymphoid Neoplasms (REAL) classification is based on the principle that each type of lymphoma is a distinct disease defined by morphology, immunophenotypic and genetic features, clinical presentation, and course. If either primary or secondary involvement of the skin is a constant factor, this aspect is considered integral to disease definition. Organ-specific classification schemes, such as that proposed by the European Organization for Research and Treatment of Cancer (EORTC) for cutaneous lymphomas, are not required, and indeed may impede the recognition of common features of diseases involving multiple anatomic sites. The use of multiple classification systems is a step backward, and may lead to confusion among hematologists/oncologists and dermatologists. Nevertheless, cutaneous lymphomas in many instances are distinct. Their natural history is often more indolent than nodal lymphomas, and for that reason they often require different therapeutic approaches. We agree with the efforts of the EORTC classification to emphasize the unique clinical aspects of many cutaneous lymphomas, as this recognition is essential for appropriate clinical management. As has been learned for nodal lymphomas, clinical features play an important role in prognosis and should be used in guiding therapy. For cutaneous lymphomas, the presence or absence of systemic spread is particularly important. PMID- 10380052 TI - Classification of cutaneous lymphoma: a critical appraisal of recent proposals. AB - The classification of cutaneous lymphoma is a contentious issue. In this short review the merits of REAL classification and EORTC classification for cutaneous lymphoproliferative disease are examined. Points of terminological confusion between the two schemes are considered and a brief account of less common or ambiguous lymphoma types is provided. PMID- 10380053 TI - A critique of classifications of lymphoma in historical perspective (and a proposal for how a classification that actually works can be formulated). PMID- 10380054 TI - Undying echoes: onomatopeia in general and medical language. PMID- 10380055 TI - About benign neoplasms with sebaceous differentiation. PMID- 10380056 TI - About the histopathology of erythema induratum-nodular vasculitis. PMID- 10380057 TI - Not all granular cell tumors show schwann cell differentiation: a granular cell leiomyosarcoma of the thumb, a case report. PMID- 10380058 TI - What's in a name? PMID- 10380059 TI - Prevention of corticosteroid-induced osteoporosis and fractures. PMID- 10380060 TI - Clinically significant pharmacokinetic drug interactions between antiepileptic drugs. AB - Pharmacokinetic interactions between antiepileptics represent a major potential complication of epilepsy treatment because drug combinations are common. This review discusses pharmacokinetic drug interactions of clinical significance involving antiepileptics and cytochrome P450 (CYP). Most commonly used antiepileptics are eliminated through hepatic metabolism, catalysed by the enzymes CYP2C9, CYP2C19 and CYP3A4 and uridine diphosphate glucuronosyltransferase (UDGPT). Antiepileptics are associated with a wide range of drug interactions, including hepatic enzyme induction and inhibition. Phenytoin, phenobarbiral, primidone and carbamazepine induce CYP and UDPGT enzymes while valproic acid inhibits them. Avoidance of unnecessary polypharmacy, selection of alternative agents with lower interaction potential and careful dosage adjustments based on serum drug concentration monitoring and clinical observation are the main methods for reducing the risks associated with these interactions. PMID- 10380061 TI - Treatment of prostate cancer. AB - Prostate cancer is the leading cause of cancer death among older men in western countries. However, controversy surrounds many issues related to this disease, particularly its most appropriate treatment, with a wide spectrum of opinions ranging from watchful waiting to aggressive therapy. Patients with newly diagnosed prostate cancer, as well as their doctors, will have to make difficult decisions regarding treatment of this disease. In this article we discuss the current available treatment options and some novel therapeutic approaches to tackling the patient with prostate cancer. PMID- 10380062 TI - Population-based investigations of drug relative clearance using nonlinear mixed effect modelling from information generated during the routine clinical care of patients. AB - Interpatient variability in drug disposition and response is a therapeutic premise, and thus evaluation and management of such variability are the basis for individualized pharmacotherapy. If the mathematical approach to determining drug doses were accurate and practical, the use of calculated doses could reduce the potential for toxicity and decrease the need for repetitious drug assays. The major strength of the population pharmacokinetics approach is that useful information can be extracted from sparse data collected during routine clinical care. Population pharmacokinetics can be defined as the study of the variability in serum drug concentrations between individuals when standard dosage regimens are administered. An approach to population pharmacokinetic data analysis has been implemented in the Nonlinear Mixed Effects Model (NONMEM) computer program. This report shows the feasibility of using a simple pharmacokinetic screen approach to estimate the population mean relative drug clearance and detecting drug-drug interaction by use of NONMEM. In clinical application of multiple trough screen or multiple peak screen, the variability of drug relative clearance within the population is assessed and a mathematical relationship between drug relative clearance and individual patient characteristics, such as age, body weight, gender, disease state or drug interaction with concomitant drug is derived. In this report I describe this approach and its application using several examples previously reported by us and others. PMID- 10380063 TI - Slowing the titration rate of tramadol HCl reduces the incidence of discontinuation due to nausea and/or vomiting: a double-blind randomized trial. AB - BACKGROUND: Ultram [tramadol hydrochloride (HCl)] is a centrally acting analgesic that is widely prescribed for the treatment of moderate to moderately severe chronic pain. Although tramadol is generally well tolerated, some patients discontinue use early in the course of treatment because of nausea and vomiting. OBJECTIVE: To investigate the effect of three initial titration rates of tramadol HCl on the incidence of discontinuation due to nausea and/or vomiting in patients who previously did not tolerate tramadol HCl. METHOD: A multicentre, outpatient, randomized double-blind study was conducted, comprised of two phases: a 14-day open-label run-in phase and a 28-day double-blind phase. In the run-in phase the dose of tramadol was titrated over 4 days to the target of 200 mg/day. Patients who discontinued tramadol HCl due to nausea and/or vomiting in the open-label phase were eligible to enter the 28-day double-blind phase after a 10-day wash out. Patients were randomized to one of three groups using a 10-, 16- or a 13-day titration schedule in order to achieve a target dosage of either 200 mg/day (10- and 16-day titration groups) or 150 mg/day (13-day titration group). The number of discontinuations due to nausea and/or vomiting in each group were compared. RESULTS: Significantly fewer patients (22%) discontinued because of nausea and/or vomiting in the 13- and 16-day titration groups compared to the 10-day group (P=0.008 and P=0.006, respectively). The time to discontinuation was also significantly delayed in the 13- and 16-day groups compared to the 10-day group (P=0.006 and P=0.007, respectively). The outcome of the 13-day titration to 150 mg/day was essentially the same as that of the 16-day titration to 200 mg/day, suggesting that this is a true rate effect rather than being dose related. CONCLUSION: This study demonstrated that a slower titration rate of tramadol HCl improves tolerability in patients who previously discontinued therapy due to nausea and/or vomiting. This study also demonstrates that the rate of titration of tramadol HCl rather than the target dose is the major determinant of tolerability. PMID- 10380064 TI - Compatibility of tirofiban HCl with dopamine HCl, famotidine, sodium heparin, lidocaine HCl and potassium chloride during simulated Y-site administration. AB - OBJECTIVE: To study the compatibility of tirofiban HCl injection 0-05 mg/ml with dopamine HCl, famotidine, sodium heparin, lidocaine HCl and potassium chloride infusion solutions during simulated Y-site administration. METHOD: Tirofiban HCl, dopamine HCl, famotidine, lidocaine HCl and potassium chloride infusions were each prepared from their respective concentrates as per current clinical preparation instructions in both 0.9% sodium chloride and 5% dextrose solutions at both the minimum and maximum concentrations normally administered. Sodium heparin premixed infusion solutions in 5% dextrose and 0-45% sodium chloride were used as-is. Tirofiban HCl solutions were combined 1:1 (simulated Y-site administration) with the dopamine HCl, famotidine, sodium heparin, lidocaine HCl and potassium chloride solutions in separate glass containers and polyvinylchloride Y-site infusion lines. Samples were held for 4 h at room temperature under ambient fluorescent light and were assayed for changes in drug content, degradation, pH, appearance and turbidity. Activity of sodium heparin solutions was measured using an aPTT coagulation assay. RESULTS: All mixtures remained clear and colourless with no visual indication of instability, i.e. precipitation. Clarity of solutions was confirmed by turbidometric analysis. There was no significant loss of drug, increase in known degradates, or appearance of unknown drug-related peaks as determined by HPLC. The activity of heparin in heparin-containing solutions remained unchanged. The pH of all test solutions remained constant. CONCLUSION: Tirofiban HCl injection 0.05 mg/ml can be co-infused by Y-site administration with dopamine HCl, famotidine, sodium heparin, lidocaine HCl and potassium chloride injection solutions. PMID- 10380065 TI - Communication regarding adverse drug reactions between secondary and primary care: a postal questionnaire survey of general practitioners. AB - GPs are not always informed that their patient suffered an adverse drug reaction (ADR) while in hospital. We have conducted a postal questionnaire survey of 270 GPs in order to elicit their views regarding provision of information from secondary care regarding ADRs. Of the 141 (52.2%) GPs that replied, 127 (90.1%) saw patients that had experienced an ADR in hospital. Of these GPs, 113 (89%) stated that they encountered instances where no record of the ADR existed in patients' discharge documentation. Where written information was absent, GPs are reliant on information given to them by patients. Of those responding, none were 'very confident' of this information, while 92 (78.6%) were 'uncertain' or 'very uncertain' of this information. A sample notification form was developed. GPs were generally satisfied with its content and 110 (82.7%) thought that patients should receive a copy. Almost all GPs (135 (97.8%)) felt that it would be appropriate to provide patients with ADR warning cards. Ensuring that patients and their carers are aware of drugs to which they may be allergic or intolerant through verbal and written methods should minimize the unnecessary risks of inadvertent re-exposure. PMID- 10380066 TI - Only relatively higher and lower Li+ ratio might predict clinical responses. AB - OBJECTIVE: To revisit the controversial issue regarding any possible relationship between Li+ ratio and clinical response with a view to explaining the contradictory reports on the subject. METHOD: Analysis of Li+ ratio and clinical response in 98 bipolar patients. The patients selected in this study were based on the following criteria: (i) fulfilment of DSM-III-R criteria for bipolar disorder; (ii) indication for long-term lithium therapy; (iii) continued lithium therapy for at least 2years; (iv) had data on lithium ratio. Clinical responses were classified into good responders and poor responders (partial responder +nonresponder). RESULTS: High lithium ratio (> or = 0.50) was associated with better clinical response (chi2 =5.80, d.f.=1, P< 0.05). In a further analysis, subjects with lithium ratios in the range (mean +1SD) or (0.50-0.67) and mean - 1SD or (0.32-0.49) showed the highest and lowest response rates. However, lithium ratios lower than 0.32 appear to be associated with a high response rate. CONCLUSION: The inconsistent results concerning the association between lithium ratio and clinical response reported since the 1970s may have been at least partially due or related to the inadequate inclusion of extreme lithium ratio data. However, this hypothesis needs to be tested in a larger, prospective study. PMID- 10380067 TI - Effects of sucrose, citric buffer and glucose oxidase on the stability of captopril in liquid formulations. AB - OBJECTIVE: To study the effects of sucrose, citric buffer and glucose oxidase on the stability of captopril in liquid formulations. METHOD: Captopril liquid formulations 1 mg/ml were prepared in various concentrations of syrup with and without citric buffer. The liquid formulations were stored in amber glass bottles at 5 degrees C. Samples were removed from these formulations at 0, 1, 2, 3, 4, 7, 10, 15, 20 and 30 days for assay of captopril by a stability-indicating high performance liquid chromatographic method. RESULTS: Our findings indicate that low concentration of citric buffer (0.03 M) had a small stabilising effect on captopril in liquid formulations containing no or low concentration of sucrose (10% w/v). However, in formulations with higher concentrations of sucrose (30 and 85% w/v, respectively), the stabilising effect was not apparent. Captopril in 1 mg/ml formulations containing 0.03 M citric buffer and 10% w/v glucose degraded rapidly in the presence of glucose oxidase. The degradation was particularly evident at higher temperature. Glucose oxidase is an antioxidant, which can extensively reduce oxygen concentration in liquid formulations. CONCLUSION: The stability of captopril is very sensitive to the presence of excipients. Many excipients can function as catalysts in its degradation. Even in the deficiency of oxygen, its degradation can also be substantial with suitable catalysts present in the liquid formulation. PMID- 10380068 TI - The use of the ASTRO-PU and ASTRO(97)-PU in the setting of budgets in English general practice. PMID- 10380069 TI - Dendritic pattern development of the honeybee antennal lobe neurons: a laser scanning confocal microscopic study. AB - The processing of odorant signals is performed, in the olfactory bulb of vertebrates or in the antennal lobe of insects, by different types of neurons which display specific morphological and functional features. The present work characterizes the morphogenesis of the main neuronal types which participate in olfactory discrimination in the adult honeybee (Apis mellifera). Neurons were stained intracellularly with Lucifer yellow at different stages of pupal development and in the adult, and imaged by laser scanning confocal microscopy. Attending to branching patterns, all pupal neurons could be attributed to morphological types previously established in the adult. Given the functional importance of intraglomerular dendritic arbors in the processing of olfactory information, the study focused on their development. The two main classes, dense and sparse intraglomerular arbors, display adultlike features as early as the second day of pupal development. However, morphometric measurements and confocal observations show that their general pattern undergoes continuous maturation processes until late pupal stages and after emergence of the adult. Among these, the results point out a pruning of dendritic branches in sparse arbors, but not in dense arbors. PMID- 10380070 TI - A diffusible factor from normal retinal cells promotes rod photoreceptor survival in an in vitro model of retinitis pigmentosa. AB - Transgenic mice expressing a dominant mutation in the gene for the phototransduction molecule rhodopsin undergo retinal degeneration similar to that experienced by patients with the retinal degenerative disease, retinitis pigmentosa (RP). Although the mutation is thought to cause photoreceptor degeneration in a cell-autonomous manner, the fact that rod photoreceptor degeneration is slowed in chimeric wild-type/mutant mice suggests that cellular interactions are also important for maintaining photoreceptor survival. To more fully characterize the nature of the cellular interactions important for rod degeneration in the RP mutant mice, we have used an in vitro approach. We found that when the retinas of the transgenic mice were isolated from the pigmented epithelium and cultured as explants, the rod photoreceptors underwent selective degeneration with a similar time course to that observed in vivo. This selective rod degeneration also occurred when the cells were dissociated and cultured as monolayers. These data indicate that the mutant rod photoreceptors degenerate when removed from their normal cellular relationships and without contact with the pigmented epithelium, thus confirming the relative cell autonomy of the mutant phenotype. We next tested whether normal retinal cells could rescue the mutant photoreceptors in a coculture paradigm. Coculture of transgenic mouse with wild-type mouse or rat retinal cells significantly enhanced transgenic rod photoreceptor survival; this survival-promoting activity was diffusible through a filter, was heat labile, and not present in transgenic retinal cells. Several peptide growth factors known to be present in the retina were tested as the potential survival-promoting molecule responsible for the effects of the conditioned medium; however, none of them promoted survival of the photoreceptors expressing the Pro23His mutant rhodopsin. Nevertheless, we were able to demonstrate that the mutant photoreceptors could be rescued by an antagonist to a retinoic acid receptor, suggesting that the endogeneous survival-promoting activity may function through this pathway. These data thus confirm and extend the findings of previous work that local trophic interactions are important in regulating rod photoreceptor degeneration in retinitis pigmentosa. A diffusible factor found in normal but not transgenic retinal cells has a protective effect on the survival of rod photoreceptors from Pro23His mutant rhodopsin mice. PMID- 10380071 TI - Modulation of dihydropyridine-sensitive calcium channels in Drosophila by a cAMP mediated pathway. AB - Drosophila has proved to be a valuable system for studying the structure and function of ion channels. However, relatively little is known about the regulation of ion channels, particularly that of Ca2+ channels, in Drosophila. Physiological and pharmacological differences between invertebrate and mammalian L-type Ca2+ channels raise questions on the extent of conservation of Ca2+ channel modulatory pathways. We have examined the role of cyclic adenosine monophosphate (cAMP) cascade in modulating the dihydropyridine (DHP)-sensitive Ca2+ channels in the larval muscles of Drosophila, using mutations and drugs that disrupt specific steps in this pathway. The L-type (DHP-sensitive) Ca2+ channel current was increased in the dunce mutants, which have high cAMP concentration owing to cAMP-specific phosphodiesterase (PDE) disruption. The current was decreased in the rutabaga mutants, where adenylyl cyclase (AC) activity is altered thereby decreasing the cAMP concentration. The dunce effect was mimicked by 8-Br-cAMP, a cAMP analog, and IBMX, a PDE inhibitor. The rutabaga effect was rescued by forskolin, an AC activator. H-89, an inhibitor of protein kinase-A (PKA), reduced the current and inhibited the effect of 8-Br-cAMP. The data suggest modulation of L-type Ca2+ channels of Drosophila via a cAMP-PKA mediated pathway. While there are differences in L-type channels, as well as in components of cAMP cascade, between Drosophila and vertebrates, main features of the modulatory pathway have been conserved. The data also raise questions on the likely role of DHP-sensitive Ca2+ channel modulation in synaptic plasticity, and learning and memory, processes disrupted by the dnc and the rut mutations. PMID- 10380073 TI - GLT1, glial glutamate transporter, is transiently expressed in neurons and develops astrocyte specificity only after midgestation in the ovine fetal brain. AB - Glutamate transport is a primary mechanism for regulating extracellular levels of glutamate in the central nervous system. GLT1, the most abundant of the known high-affinity glutamate transporters, is found exclusively in astrocytes in adult brain of several species, but we and others have recently identified neurons that transiently express GLT1 protein in the developing brain. We now demonstrate the development of cell type specificity for GLT1 expression at 60, 71, and 136 days' gestation in the developing sheep brain (term = 145 days). At 60 and 71 days of gestation, GLT1 colocalizes with calbindin in Purkinje cells in the cerebellum, and this expression pattern has a novel distribution that is reminiscent of the parasagittal zebrin-like bands. GLT1 immunoreactivity simultaneously occurs in periventricular white matter, anterior commissure, and striatal white matter, dissipating by 136 days. GLT1 protein expression within astrocytes is developmentally regulated, appearing first in vimentin positive radial glia at 60 and 71 days and then switching to GFAP positive parenchymal and perivascular astrocytes at 136 days. Expression of GLT1 in subsets of vimentin-positive astrocytes persists in white matter but not in cortex. These results identify a novel compartmentation within cerebellar cortex and neuronal and axonal pathway localization of GLT1, suggesting the participation of this glutamate transporter in the development of the topographic organization of cerebellar cortex and a transient neuronal function for GLT1 in developing brain. In addition, GLT1 expression is highly plastic, being neither exclusively astroglial nor uniformly expressed in different populations of astrocytes during brain development. PMID- 10380072 TI - Neurons differentiating from murine neural crest in culture exhibit sensory or sympathetic-like calcium currents. AB - The trunk neural crest gives rise to peripheral sensory and sympathetic neurons. In culture, neural crest cells can be induced to differentiate into either neuronal phenotype. Few studies have examined the differentiation of physiological properties in cultures of neural crest cells. Using whole-cell recordings, our study examined the effects of growth factors on high-voltage activated calcium current profiles exhibited by neurons differentiating in culture. We compared these profiles with those exhibited by sensory and sympathetic neurons. Neural crest cells in culture gave rise to neurons with calcium current profiles identical to either sensory or sympathetic neurons, depending on the growth conditions. On average, the calcium current profile for sensory neurons was 23% (L), 51% (N), and 12% (P), while sympathetic neurons had a similar L-type current (20%), higher N-type (76%), and lower P-type (4%). Neural crest cells cultured with human leukemia inhibitory factor plus somite cells produced neurons with a sympathetic-like calcium current profile (L: 17%, N: 75%, and P: 4%). However, murine leukemia inhibitory factor (L: 25%, N: 52%, and P: 13%) and ciliary neurotrophic factor (L: 18%, N: 49%, and P: 9%) plus somite cells produced neurons with sensory-like calcium current profiles. These growth conditions did not modify the calcium current profiles of neurons cultured from embryonic and neonatal ganglia. Similarly, murine leukemia inhibitory factor produced a greater percentage of neurons (57%) with sensitivity to capsaicin (sensory phenotype) than human leukemia inhibitory factor (3%). Physiological traits can be a useful tool for the determination of neuronal phenotype in culture where other traits may be less stable. PMID- 10380074 TI - Proliferation of vertebrate inner ear supporting cells. AB - Using two S phase markers, we determined the cell-cycle behavior of inner ear supporting cells from two species, the chicken and the oscar. The results indicate that chicken utricular supporting cells divide once and do not return to the cell cycle for at least 7 days. In contrast, supporting cell progeny in the oscar saccule return to S phase after 5 days. While both the chicken utricle and oscar saccule show ongoing supporting cell proliferation, these data indicate that there may be a dedicated recycling population of supporting cells in the oscar saccule but not in the chicken utricle that is responsible for hair cell production. An expulsion of proliferative cell progeny in the chicken utricle after 7 days may be a driving force for proliferation, as well as an explanation for why hair cell numbers do not increase in the chicken utricle with age. This was not seen in the oscar saccule, possibly explaining how this end organ increases in size throughout the adult life of the animal. The absence of S phase cell expulsion, however, does not rule out the role of cell death in the oscar saccule. PMID- 10380075 TI - Phosphatidylinositol 3-kinase-mediated regulation of neuronal apoptosis and necrosis by insulin and IGF-I. AB - We examined effects of two insulin-like growth factors, insulin and insulin-like growth factor-I (IGF-I), against apoptosis, excitotoxicity, and free radical neurotoxicity in cortical cell cultures. Like IGF-I, insulin attenuated serum deprivation-induced neuronal apoptosis in a dose-dependent manner at 10-100 ng/mL. The anti-apoptosis effect of insulin against serum deprivation disappeared by addition of a broad protein kinase inhibitor, staurosporine, but not by calphostin C, a selective protein kinase C inhibitor. Addition of PD98059, a mitogen-activated protein kinase kinase (MAPKK) inhibitor, blocked insulin induced activation of extracellular signal-regulated protein kinases (ERK1/2) without altering the neuroprotective effect of insulin. Cortical neurons underwent activation of phosphatidylinositol (PI) 3-kinase as early as 1 min after exposure to insulin. Inclusion of wortmannin or LY294002, selective inhibitors of PI 3-K, reversed the insulin effect against apoptosis. In contrast to the anti-apoptosis effect, neither insulin nor IGF-I protected excitotoxic neuronal necrosis following continuous exposure to 15 microM N-methyl-D-aspartate or 40 microM kainate for 24 h. Surprisingly, concurrent inclusion of 50 ng/mL insulin or IGF-I aggravated free radical-induced neuronal necrosis over 24 h following continuous exposure to 10 microM Fe2+ or 100 microM buthionine sulfoximine. Wortmannin or LY294002 also reversed this potentiation effect of insulin. These results suggest that insulin-like growth factors act as anti apoptosis factor and pro-oxidant depending upon the activation of PI 3-kinase. PMID- 10380076 TI - Formation of functional synaptic connections between cultured cortical neurons from agrin-deficient mice. AB - Numerous studies suggest that the extracellular matrix protein agrin directs the formation of the postsynaptic apparatus at the neuromuscular junction (NMJ). Strong support for this hypothesis comes from the observation that the high density of acetylcholine receptors (AChR) normally present at the neuromuscular junction fails to form in muscle of embryonic agrin mutant mice. Agrin is expressed by many populations of neurons in the central nervous system (CNS), suggesting that this molecule may also play a role in neuron-neuron synapse formation. To test this hypothesis, we examined synapse formation between cultured cortical neurons isolated from agrin-deficient mouse embryos. Our data show that glutamate receptors accumulate at synaptic sites on agrin-deficient neurons. Moreover, electrophysiological analysis demonstrates that functional glutamatergic and gamma-aminobutyric acid (GABA)ergic synapses form between mutant neurons. The frequency and amplitude of miniature postsynaptic glutamatergic and GABAergic currents are similar in mutant and age-matched wild type neurons during the first 3 weeks in culture. These results demonstrate that neuron-specific agrin is not required for formation and early development of functional synaptic contacts between CNS neurons, and suggest that mechanisms of interneuronal synaptogenesis are distinct from those regulating synapse formation at the neuromuscular junction. PMID- 10380077 TI - Developmental regulation of the neuronal-specific isoform of K-Cl cotransporter KCC2 in postnatal rat brains. AB - We examined the expression of the KCC2 isoform of the K-Cl cotransporter in the developing and adult brain, using an affinity-purified antibody directed against a unique region of the KCC2 protein. Expression was shown to be limited to neurons at the cell bodies and cell processes in the hippocampus and cerebellum. Expression seemed to be the highest at the end of processes that originated from the CA1 pyramidal cells. Developmental up-regulation of KCC2 expression was demonstrated in the entire rat brain by Northern and Western blot analyses, and in the hippocampus by immunofluorescence. Level of KCC2 expression was minimal at birth and increased significantly during postnatal development. This pattern of expression was opposite to the one of the Na-K-2Cl cotransporter that is highly expressed in immature brain and decreases during development. The up-regulation of the K-Cl cotransporter expression is consistent with the developmental down regulation of the intracellular Cl- concentration in neurons. The level of intracellular Cl-, in turn, determines the excitatory versus inhibitory response of the neurotransmitter gamma-aminobutyric acid in the immature versus mature brain. Finally, KCC2 expression was shown in dorsal root ganglion neurons, demonstrating that expression of the cotransporter is not strictly confined to central nervous system neurons. PMID- 10380078 TI - Enriched environment increases neurogenesis in the adult rat dentate gyrus and improves spatial memory. AB - The fetal and even the young brain possesses a considerable degree of plasticity. The plasticity and rate of neurogenesis in the adult brain is much less pronounced. The present study was conducted to investigate whether housing conditions affect neurogenesis, learning, and memory in adult rats. Three-month old rats housed either in isolation or in an enriched environment were injected intraperitoneally with bromodeoxyuridine (BrdU) to detect proliferation among progenitor cells and to follow their fate in the dentate gyrus. The rats were sacrificed either 1 day or 4 weeks after BrdU injections. This experimental paradigm allows for discrimination between proliferative effects and survival effects on the newborn progenitors elicited by different housing conditions. The number of newborn cells in the dentate gyrus was not altered 1 day after BrdU injections. In contrast, the number of surviving progenitors 1 month after BrdU injections was markedly increased in animals housed in an enriched environment. The relative ratio of neurogenesis and gliogenesis was not affected by environmental conditions, as estimated by double-labeling immunofluorescence staining with antibodies against BrdU and either the neuronal marker calbindin D28k or the glial marker GFAp, resulting in a net increase in neurogenesis in animals housed in an enriched environment. Furthermore, we show that adult rats housed in an enriched environment show improved performance in a spatial learning test. The results suggest that environmental cues can enhance neurogenesis in the adult hippocampal region, which is associated with improved spatial memory. PMID- 10380079 TI - Growth cone neuropilin-1 mediates collapsin-1/Sema III facilitation of antero- and retrograde axoplasmic transport. AB - Collapsin-1/Sema III, a member of the semaphorin family, has been implicated in axonal pathfinding as a repulsive guidance cue. Cellular and molecular mechanisms by which collapsin-1 exerts its action are not fully understood. Collapsin-1 induces growth cone collapse via a pathway which may include neuropilin-1, a cellsurface collapsin-1 binding protein, as well as intracellular CRMP-62 and heterotrimeric G proteins. We previously identified a second action of collapsin 1, the facilitation of antero- and retrograde axoplasmic transport. This response occurs via a mechanism distinct from that causing growth cone collapse. To investigate the possible involvement of neuropilin-1 in the action of collapsin-1 on axoplasmic transport, we produced a soluble neuropilin-1 (sNP-1) lacking the transmembrane and intracellular region. sNP-1 progressively displaced the dose response curve for collapsin-1 to induce growth cone collapse to higher concentrations. sNP-1 also inhibited collapsin-1-induced augmentation of both antero- and retrograde axoplasmic transport. Furthermore, an anti-neuropilin-1 antibody blocked the collapsin-induced axoplasmic transport. These results together indicate that neuropilin-1 mediates collapsin-1 action on axoplasmic transport. To visualize collapsin-1 binding to endogenous neuropilin-1, we used a truncated collapsin-1-alkaline phosphatase fusion protein (CAP-4). CAP-4 stains the growth cone, neurite, and cell body. However, local application of collapsin 1 to growth cone but to neither neurite nor cell body promotes axoplasmic transport. Thus, growth cone NP-1 mediates the facilitatory action of collapsin-1 on antero- and retrograde axoplasmic transport. PMID- 10380080 TI - A dual-probe fluorescence method to examine selective perturbations of membrane permeability by melittin. AB - A new fluorescence method has been developed to measure simultaneously and independently the release of fluorophores from two vesicle populations. Calcein and sulforhodamine B were used as a probe couple: the leakage of these probes from vesicles can be recorded independently since they can be excited simultaneously at 510 nm, and their individual fluorescence can be isolated by measuring the fluorescence signal at 525 and 590 nm, using a T-shape fluorometer. Controls show that both probes are suitable for the leakage assay based on fluorescence self-quenching, that they do not interact physically or chemically at the concentrations used in the method, and that they leak in a similar fashion from a given vesicle type. This dual-probe technique is applied to examine the specificity of the release relative to the cholesterol content of the vesicles for melittin, a toxin. This new approach shows in a straightforward manner that melittin-induced release for a given population can be modulated by the presence of vesicles with another lipid composition and this competitive release is associated with a preferential distribution of the peptide on the targeted vesicles. PMID- 10380081 TI - Protonation of porphyrin in iron-free cytochrome c: spectral properties of monocation free base porphyrin, a charge analogue of ferric heme. AB - Charged groups reside mainly on protein surfaces, but for proteins that incorporate redox centers, a charge typically exists at the prosthetic group within the interior. How a protein accommodates a buried charge and the effect of redox changes on protein stability are thermodynamically related problems. To examine these problems in cytochrome c, the metal-free protein was used as a model. When pH is lowered, the neutral, monocation, and dication forms of the porphyrin are progressively formed as indicated by their characteristic absorption spectra. Infrared studies of the protein over this pH range show that the protein remains in a predominately alpha-helical structure, although the carboxyl groups of the dicarboxylic amino acids become protonated at lower pH. The monocation porphyrin form (which has not been previously reported in a protein and is a charge analogue of ferric heme) has a fluorescence maximum at 609 nm. The pKs for the respective one and two protonation of the porphyrin pyrrole Ns are 3.2 and 1.6 for the folded protein, and 4.4 and 3.1 for the unfolded protein. These values indicate that the protection of the polypeptide chain for protonation is approximately 3 kcal. PMID- 10380082 TI - Lipid composition, membrane structure relationships in lens and muscle sarcoplasmic reticulum membranes. AB - Membrane lipid composition varies in different tissues and species. Since a defined lipid composition is essential to the function of many membranes, the relationship between membrane lipid composition and structure was determined using infrared and Raman spectroscopy in four membranes containing a calcium pump: rabbit fast and slow twitch muscle sarcoplasmic reticulum and human and bovine lens fiber cell membranes. We found that membrane sphingolipid and phosphatidylcholine content were correlated to a decrease and increase, respectively, in the infrared lipid CH2 symmetric stretching band frequency. We interpret the change in frequency as a change in lipid hydrocarbon chain structural order. This was confirmed by Raman order parameters. The high degree of hydrocarbon chain saturation found in the variable amide chains of sphingolipids is likely to account for this correlation. Lipid phase transition temperature and cooperativity also correlated to sphingolipid and phosphatidylcholine content, and are the forces defining the order in at physiological temperature in the samples studied. Ca(2+)-ATPase caused an increase in the CH2 symmetric stretching frequency in fast twitch muscle sarcoplasmic reticulum (interpreted as an increase in hydrocarbon chain disorder), but had no effect on slow twitch muscle sarcoplasmic reticulum lipid hydrocarbon chain structure. In the natural systems studied, we find that it is the lipid hydrocarbon chain saturation that defines lipid hydrocarbon chain order. PMID- 10380083 TI - Separable contributions of ordered and disordered lipid fatty acyl chain segments to nuCH2 bands in model and biological membranes: a Fourier transform infrared spectroscopic study. AB - In this article, the assignment of the nu(C-H) stretching region of lipid molecules is revisited. This region is extensively used to follow lipid phase transitions, and especially the frequency shifts and bandwidth alterations in the nu(sym)CH2 band have been utilized in this respect. Here, we propose and prove that behind these phenomena there are pairs of component bands in the cases of both the nu(sym)CH2 and the nu(as)CH2 bands. The lower-frequency components of the pairs are assigned to the vibrations of CH2 groups on trans segments of the fatty acyl chains, while the higher-frequency components of the pairs are assigned to CH2 groups on gauche segments. To prove these assignments, we have shown that the nuCH2 frequencies are characteristic of the conformation of the lipid fatty acyl chain itself, and not the state of the whole lipid matrix. Curve fitting in fact revealed the conformer-specific components. With the use of singular value decomposition analysis we have demonstrated that the relative intensity changes in the components, and not the shifts in the whole bands, cause the observed shifts in the nuCH2 bands upon lipid phase transition. The results of this approach are presented for deuterium-saturated dioleoyl phosphatidylcholine mixtures, for the gel --> liquid-crystalline phase transition of dipalmitoyl-phosphatidylcholine multilayers, and for a biological membrane, barley thylakoid. This refined assignment offers physically plausible reasoning for the observed phenomena and is able to explain frequency shifts and bandwidth changes observed previously upon lipid phase transitions, including their nonconcerted temperature dependences. In biological membranes, this interpretation allows the separation of protein- and membrane-dynamics-induced lipid conformational changes. PMID- 10380084 TI - 5,10,15,20-Tetrakis(4-N-methylpyridyl)porphyrinato-palladium(II ) as a differentiation probe for sensing binding modes with B-DNA duplexes: electronic MCD and CD spectra. AB - We report our detailed electronic MCD, CD, and optical spectroscopic measurements and analysis of the porphyrin Soret (B(o)) region of four-coordinate 5,10,15,20 tetrakis(4-N-methylpyridyl)porphyrinatopalladium( II), PdP(4), and its bound states with B-DNA duplexes poly(A-T)2, poly(G-C)2, and calf thymus DNA (CT DNA). For system PdP(4)/poly(A-T)2 it was possible to conclude that the porphyrin is bound edge-on in the major groove, specifically at the 5'AT3' site. For this orientation the porphyrin's electric dipole transition moments (edtm), mu(x) (most perturbed direction) and mu(y) (least perturbed direction), have tilt angles alpha approximately 90 degrees and approximately 45 degrees , respectively, relative to the helix axis. It was further concluded from the small shifts of B(o) optical and MCD band intensities and wavelengths and from the net MCD (+) A-term sign retention upon binding that the porphyrin's frontier ppi MOs (1a(1u) 3a(2u) 4eg) are only weakly perturbed by the heterocyclic bases of poly(A T)2, and therefore that the LUMO (4eg) splitting is less than the |1a(1u)-3a(2u| energy separation, deltaHOMO, that is, deltaLUMO < deltaHOMO for the bound state in PdP(4)/poly(A-T)2. For intercalation systems PdP(4)/ poly(G-C)2 and /CT DNA, with PdP(4) centered in the intercalation "pocket" and having two of its 4-N methylpyridyls extending into each of the major and minor grooves, the edtms mu(x) and mu(y) were determined to be oriented perpendicular (gamma approximately 0 degrees) and parallel (gamma approximately 90 degrees) to the hydrogen bonds of the base pairs, respectively. Intercalation is characterized by a much stronger binding interaction, viz., the B(o) optical band and net MCD extrema wavelength shifts are relatively large, and the net MCD (+) A-term of PdP(4) is substantially quenched as it becomes the (-) pseudo-A-term of intercalated PdP(4)/poly(G-C)2. This A-term sign reversal informs that the porphyrin MOs are so strongly perturbed by the GC base pairs that deltaLUMO > deltaHOMO, which gives rise to a (-) pseudo-A-term. Also, the findings demonstrate (1) the potential of PdP(4) as a sensitive, discriminating analytical probe of DNA sequences and (2) the diagnostic capability of the composite of five spectra [net MCD, CD, and optical of free and bound PdP(4)] in differentiating the site and sequence selectivity and preferred binding mode of this porphyrin. PMID- 10380085 TI - Circular dichroism and molecular modeling of the E. coli TolA periplasmic domains. AB - Colicins are killer proteins that use envelope proteins from the outer and the inner membranes to reach their cellular target in susceptible cells of Escherichia coli. Each group A colicin uses a combination of Tol proteins to cross the outer membrane of gram-negative bacteria and to exert their killing activity. The TolA protein, necessary for the import of all the group A colicins, is a 421-amino acid residue protein composed of three domains (TolAI, TolAII, and TolAIII). TolAIII interacts with the N-terminal domain of colicin A (AT1). Analytical ultracentrifugation reveals that TolAII and TolAIII are monomer structures, TolAII has an elongated structure, and TolAIII is rather globular. Circular dichroism (CD) spectra were done with TolAII-III, TolAII, TolAIII, AT1, and the AT1-TolAII-III complex. TolA CD spectra reveal the presence of alpha helix structure in aqueous solution and the intensity of the a-helix signal is the highest with TolAII. Few structural changes are observed with the complex AT1 TolAII-III. Molecular modeling was done for TolAII-III, taking into account CD and ultracentrifugation data and show that domain II can adopt a barrel structure made of three twisted alpha-helices similar to coiled coil helices while domain III can adopt a globular structure. PMID- 10380086 TI - Inhaled corticosteroids in childhood asthma. PMID- 10380087 TI - Does treatment of asthmatic children with inhaled corticosteroids affect their adult height? AB - In this retrospective study, adult height was assessed in young adult asthmatics who were treated with inhaled corticosteroids (ICs) during childhood (n = 42; 26 boys) and compared to those obtained in asthmatic patients who were never treated with ICs during childhood (n = 43; 23 boys). Standing height of all subjects and their parents was measured. Height data were analyzed using actual length and target height in centimeters, standard deviation scores (SDS), and difference between adult height of the patients and their target height (adult height minus target height). Mean adult height was the same in subjects who took ICs during childhood as compared to those who had never received ICs (boys: 179.3cm+/-6.8 vs. 180.4 cm+/-5.6; girls: 165.8 cm+/-7.5 vs. 167.7 cm+/-7.2). SDS of adult height was also not different between the two groups: in subjects who did not take ICs it was 0.89+/-1.00, while in those who took ICs it was 0.66+/-1.10 (P = 0.31). SDS of target height was also not different between the two groups: in subjects not taking ICs it was 0.95+/-0.86, while in those who took ICs it was 0.28+/-0.76 (P = 0.30). However, subjects who took ICs during childhood showed a statistically significant lower value of adult height minus target height than those who never took ICs (whole group: -0.003+/-5.9 vs. 2.54 +/-4.8, P = 0.03 ; boys: 0.004+/-5.8 vs. 3.09+/-4.5, P = 0.04 ; girls: -0.075+/-6.3 vs. 1.91+/-5.2, P = 0.31). Patients on ICs during childhood who had ever been hospitalized for asthma showed a lower value for adult height minus target height than those who took ICs but were never hospitalized (-3.08+/-7.8 vs. 1.06+/-4.8, P = 0.046). A logistic regression analysis predicting growth impairment showed that the best fitting model was one that used only ICs as a dependent variable (crude odds ratio, 3.3; 95% CI, 1.3-8.4). Patients who were treated with ICs in combination with intranasal corticosteroids (treatment for rhinitis) tended to have a lower value of adult height minus target height than the other children, but the difference was not statistically significant (P = 0.07). We conclude that although adult height was the same in young adults who were treated with ICs during childhood compared to those who were not treated with ICs during childhood, there was a statistically significant difference between the two groups for adult height minus target height, suggesting mild growth retardation in patients who took ICs during childhood. These findings may be explained by the use of ICs, but it seems more likely that a difference in asthma severity between both groups was responsible for it. PMID- 10380088 TI - Early effects of inhaled steroids on airway hyperreactivity and pulmonary function in asthma. AB - While inhaled steroids (IS) are increasingly recognized as having a more rapid onset of action than was once thought, little is known about the early changes in objective measures of respiratory function that follow the inhalation of repeated doses. These early effects were examined in a randomized, double-blind, placebo controlled, crossover study of 20 children aged 10-16 years with stable mild asthma. Beclomethasone dipropionate (BDP) 2,000 mcg, fluticasone propionate (FP) 400 mcg, and placebo were given twice daily for three doses. Airway hyperreactivity (AHR) to methacholine (PC20), pulmonary function tests (PFT: FVC, FEV1, FEF25-75%), and the rate of recovery from methacholine-induced bronchospasm following administration of salbutamol were determined at 8 h (after 1 dose) and at 32 h (after three doses). At 8 h, minor improvements in AHR were demonstrated, averaging 0.32 doubling doses in PC20. At 32 h, significant improvements in AHR and PFTs were present, averaging 0.92 doubling doses in PC20, 3.96% of predicted values in FEV1, and 7.74% of predicted values in FEF25-75%. No significant changes occurred in FVC. There were no significant differences between the effects of BDP and FP. Inhaled steroids were associated with a slower response to salbutamol following methacholine challenge testing at 32 h. We conclude that IS, given in repeated high doses, result in significant improvements within 32 h in both AHR and PFTs, along with changes in response to beta2 agonists. These effects are likely to be the result of the topical activity of IS. PMID- 10380089 TI - Effects of inhaled beclomethasone compared to systemic dexamethasone on lung inflammation in preterm infants at risk of chronic lung disease. AB - The purpose of this study was to compare the effects of daily inhaled beclomethasone (3 x 500 microg) started on day 3 of life, with that of systemic dexamethasone (0.5 mg/kg/day) started between days 11-13 on clinical variables, lung inflammation, and pulmonary microvascular permeability in preterm infants at risk for chronic lung disease (CLD). Following administration of surfactant, preterm neonates with RDS and a birth weight of less than 1,200 g were included in this comparative observational pilot study when still mechanically ventilated and with an oxygen requirement on the third day of life. The patients (gestational age 26.1+/-0.9 weeks, birth weight 826+/-140 g, mean+/-SD) were alternately allocated to prophylactic treatment with inhaled beclomethasone (n = 7), or to early systemic dexamethasone therapy after day 10 of life, if clinically indicated (n = 9). Pulmonary inflammation and lung permeability were assessed by analyzing the levels of interleukin-8, elastase alpha1 proteinase inhibitor, free elastase activity, and albumin in tracheal aspirates on days 10 and 14 of life. The secretory component of IgA served as reference protein. We observed no significant differences in the concentrations of interleukin-8, elastase alpha1 proteinase inhibitor, and albumin between the two groups on day 10 of life. On day 14, 3 (median; range, 1-3) days following initiation of dexamethasone treatment, concentrations of the inflammatory mediators and of albumin were significantly lower in the group on systemic steroid therapy than in the group treated with inhaled steroids (P < 0.01). Additionally, there was a significant difference in oxygen requirements between both groups on day 14. In the group treated with inhaled steroids, concentrations of the inflammatory mediators, albumin, and oxygen requirements did not show a difference between day 10 and 14. We conclude that, in contrast to systemic dexamethasone treatment, a 12-day course of inhaled beclomethasone does not affect lung inflammation and pulmonary microvascular permeability in preterm infants at risk for CLD within the first 2 weeks of life. PMID- 10380090 TI - Early prediction of neonatal chronic lung disease: a comparison of three scoring methods. AB - A variety of postnatal therapies have been and will be evaluated for prevention or treatment of neonatal chronic lung disease (CLD). A simple method for early selection of the highest risk infants would optimize intervention trials. Our study compared a clinical scoring system for predicting neonatal CLD (defined at 36 weeks postconceptional age) with previous regression models developed by Sinkin et al. (Sinkin model) [Pediatrics 1990;86:728-736] and Ryan et al. (Ryan model) [Eur J Pediatr 1996;668-671] in two distinct populations. A respiratory failure score (RFS) was prospectively developed for infants at <32 weeks of gestation admitted to the Wilford Hall Medical Center from January 1990-December 1992. Logistic regression modeling identified three independent predictors of CLD: gestation, birth weight, and RFS. Applying a modified RFS score (to include gestation and birth weight), the RFS, Sinkin, and Ryan models were compared among high-risk infants admitted to Wilford Hall from January 1993-December 1995, and to Crawford Long Hospital (Atlanta, GA) from January 1993-December 1994. Predictive values, sensitivity, specificity, and receiver operating characteristic (ROC) curves were determined for the primary outcome variable: CLD at 36 weeks of corrected gestation. Of 248 infants at <32 weeks admitted to Wilford Hall, 220 survived >7 days. Thirty of 31 (97%) infants diagnosed with CLD were <29 weeks or < or =1,000 g at birth. Despite important demographic and treatment differences between the study populations, similar ROC curves were found for each scoring method when individually evaluated among the three study groups. The RFS method at 72 h demonstrated the greatest area under the ROC curve for prediction of neonatal CLD in the groups as a whole. Application of the RFS method for early prediction of neonatal CLD at age 72 h should improve patient selection for early prevention trials. PMID- 10380092 TI - Clinical features of obstructive sleep apnea hypoventilation syndrome in otherwise healthy children. AB - Obstructive sleep apnea hypoventilation syndrome (OSAHS) is an important public health problem. However, major gaps exist in our knowledge about the clinical features of this disorder in the pediatric age group. The purpose of this study was to examine clinical features of OSAHS diagnosed by polysomnography in otherwise healthy children. In this cross-sectional study, 326 children without underlying medical conditions (5.8+/-3.0 years, range 1-12 years; 56% male) were recruited from patients referred by primary care and otorhinolaryngology physicians for evaluation of snoring and difficulty breathing. Ethnic group distribution was African-American (38%), Caucasian (30%), and Hispanic (31%). Complaints of daytime tiredness or sleepiness were reported in 29% of the children. All children underwent overnight polysomnography (N = 330 studies). OSAHS was diagnosed in 59% of the children, based on polysomnographic criteria. The remaining children had either primary snoring (25%); no snoring (10%), or upper airway resistance syndrome (6%). Neither male gender nor obesity increased the likelihood for the diagnosis of OSAHS. However, the incidence of obesity in the study population (28%) was more than twice that of the general pediatric population. African-American children had a greater likelihood for OSAHS diagnosis compared to Hispanic or Caucasian children. Daytime complaints of sleepiness or tiredness were not more common in children diagnosed with OSAHS than in the children without OSAHS. As expected, tonsillar hypertrophy increased the likelihood of OSAHS diagnosis. In summary, many of the clinical features of childhood OSAHS are in marked contrast to those in adults. PMID- 10380091 TI - Bronchoalveolar lavage in children with chronic diffuse parenchymal lung disease. AB - The aim of the present study was to compare cellular and noncellular components of bronchoalveolar lavage fluid (BAL) in a group of children with a diagnosis of chronic diffuse parenchymal lung disease (cDPLD) and a group of children without parenchymal lung disease undergoing BAL for various clinical indications (control group). We evaluated cellular and non-cellular components (total proteins, albumin, hyaluronic acid, and fibronectin) in BAL fluid from 14 children (7 boys and 7 girls; mean age 9.2 years, range 5 months to 18.4 years) fulfilling the clinical and radiological diagnosis of chronic cDPLD, and in 19 controls without evidence of lung disease. The 14 patients were assigned to two study groups: early-stage cDPLD (6 patients; age range 5 months to 5.2 years; duration of illness, 5-7 months) and long-standing cDPLD (8 patients; age range 9.6-18.4 years; duration of illness, 1.2-17.6 years). Ninety-three percent of the patients with cDPLD had at least two BAL constituents outside normal limits, with high numbers of cells, including all types of alveolar cells, but especially lymphocytes and foamy macrophages. These findings indicate a mixed, predominantly lymphocytic alveolitis. Our patients also had a significant increase in two noncellular BAL components, namely fibronectin and hyaluronic acid. BAL samples from children with long-standing cDPLD contained increased numbers of lymphocytes, whereas samples from children with early-stage cDPLD contained increased percentages and numbers of foamy macrophages and increased concentrations of fibronectin, hyaluronic acid, and albumin. In conclusion, we clearly identified an abnormal BAL profile in our group of cDPLD patients. Moreover, BAL findings differentiated younger cDPLD patients in the early stages of their illness from old patients with long-standing disease. PMID- 10380093 TI - Effect of single versus multiple dosing on perfluorochemical distribution and elimination during partial liquid ventilation. AB - The objective of this study was to quantitate perfluorochemical (PFC) elimination kinetics during partial liquid ventilation (PLV) following an initial fill with or without hourly dosing. Young New Zealand rabbits were studied in two groups: Gr I (n = 6), PLV with a single dose of PFC liquid (perflubron: LiquiVent, Alliance Pharmaceutical Corp.); and Gr II (n = 5), PLV with PFC liquid and multiple hourly dosing . All rabbits were studied for 4 h, following initial instillation of a volume of PFC liquid equal to the measured gas functional residual capacity. Animals were ventilated at a constant breathing frequency (30 br/min), tidal volume (9.3+/-0.3 SE mL/kg), positive end expiratory pressure (4 cm H2O), and inspiratory time (0.30 s). PFC saturation of mixed expired gas (PFC Sat) was assessed with a thermal conductivity analyzer, and PFC elimination was calculated from PFC-Sat, minute ventilation, and temperature of the expired gas. In GR II, PFC was supplemented hourly at a volume determined by PFC elimination calculations. The results demonstrated a decrease in PFC-sat and PFC loss with time, independent of group (P< 0.05). In addition, with hourly supplementation (GR II), PFC-Sat and PFC elimination over time was significantly (P < 0.05) greater than in animals (GR I) which did not receive additional doses. These data demonstrate that the PFC elimination rate is not constant and is related to the amount of PFC in the respiratory system. This may have occurred due to distributional differences of ventilation and PFC liquid between the single and multiple dosing groups. These findings also suggest that evaluation of PFC concentrations in expired gas may be a clinically useful index of intrapulmonary PFC distribution during PLV, and that maintained elevation of expired gas PFC saturation may guide optimal PFC dosing intervals and distribution to maximize protection against barotrauma. PMID- 10380094 TI - Lipid-laden macrophages in bronchoalveolar lavage fluid as a marker for pulmonary aspiration. AB - An increased lipid content in alveolar macrophages of bronchoalveolar lavage (BAL) fluid is thought to be a useful indicator for recurrent pulmonary aspiration. To assess whether pulmonary diseases unrelated to aspiration can raise the lipid content in alveolar macrophages, we evaluated Oil-Red-O-stained smears of BAL fluid in 18 children aged 3-15 years undergoing elective surgery for nonpulmonary illnesses under general anesthesia and in 18 children aged 1-16 years who had pulmonary diseases without clinical evidence of aspiration (pneumonia, exogenous allergic alveolitis, or cystic fibrosis). A semiquantitative lipid-laden macrophage (LLM) index was determined for each patient. LLM indices in children without pulmonary disease were higher than those published for healthy adults. In children with pulmonary diseases but without evidence of aspiration, a significantly higher LLM index was observed compared to controls. The LLM indices of children with pulmonary diseases were similar to those published by other authors for children with pulmonary aspiration. We conclude that an elevated LLM index in alveolar macrophages of BAL can be found in a variety of pulmonary diseases in which there is no clinical evidence of aspiration and is therefore unlikely to be a specific parameter for silent pulmonary aspiration. PMID- 10380095 TI - Relation between pulse oximetry and clinical score in children with acute wheezing less than 24 months of age. AB - The aim of this study was to determine the relation between transcutaneous hemoglobin oxygen saturation, measured by pulse oximetry (SpO2), and clinical score values in 138 infants (mean+/-SD, 6.6+/-5.5 months of age) with acute wheezing episodes presenting in a primary care outpatient setting. A single investigator evaluated the severity of the acute wheezing episodes by assigning a clinical score and was unaware of the SpO2 values. Another investigator measured SpO2 values on all subjects. The mean (+/-SD) SpO2 value was 98.2+/-1.1% for children with clinical scores of 2-5 (n = 32); 95.4+/-1.5% for those with scores of 6-7 (n = 82), and 92.9+/-2% for children with scores of 8-10 (n = 24), (P < 0.001 by Bonferroni's multiple comparison, when all two-way comparisons were done for each pair of results). The clinical score showed a good correlation with SpO2 (r = -0.76; 95% CI, -0.83 to -0.68). We conclude that if pulse oximetry is not available, it is advisable to include oxygen in the therapy of wheezy infants with clinical scores values >8. PMID- 10380096 TI - Assessment of a peak flow whistle in nonasthmatic children. AB - We tested the agreement of peak expiratory flow (PEF) measurements between an electronic spirometer and a peak flow whistle (Whistle Watch, HarMed, Capetown, South Africa). One hundred and three healthy children between ages 6-13 years and with no previous experience in lung function tests participated in the study. Sequential PEF-readings were obtained from the spirometer and the peak flow whistle; all children had an equal number of attempts using both devices. In the case of the spirometer, the highest PEF reading of three acceptable and reproducible efforts was noted as the best PEF (PEF(SPIRO)). Whistle Watch readings were taken as the highest value when the child could activate the whistle. Despite a strong correlation (r = 0.91; R2 = 83%) between the readings of the spirometer and Whistle Watch, there was a lack of agreement between the two devices. For any individual subject, the 95% probability interval ranged between +30.4 to -47 L.min(-1); 64% of the children obtained higher PEF-values on Whistle Watch, compared to the spirometer. These findings suggest that the whistle sound of the peak flow whistle was a significant incentive, which resulted in greater maximal expiratory efforts. PMID- 10380097 TI - Nosocomial transmission of penicillin-resistant Streptococcus pneumoniae. AB - Two patients were found to harbor intermediate-level penicillin-resistant Streptococcus pneumoniae in a pediatric hospital setting. For the first patient, the bacterium was isolated from a tracheal aspirate, and for the second patient, a positive blood culture was found a short time after the index case. Two molecular typing techniques (enterobacterial repetitive intergenic consensus sequence polymerase chain reaction, and repetitive sequence-based polymerase chain reaction) demonstrated homology among these isolates, which suggests person to-person spread. We propose the need for institution-based infection control precautions that will limit the spread of penicillin-resistant pneumococci. PMID- 10380098 TI - Diversity of expressed cytochrome P450 genes in the adult Mediterranean Fruit Fly, Ceratitis capitata. AB - The cytochrome P450s comprise a superfamily of mostly microsomal haemoproteins which play a dominant role in the metabolism of a variety of endogenous and foreign compounds. The use of a degenerate PCR primer targeted to the haem binding decapeptide unique to the cytochrome P450 superfamily resulted in the identification of 14 novel cytochrome P450s in the Mediterranean Fruit Fly, Ceratitis capitata. Analysis of the relative frequency of individual isoforms within the pool of isolated sequences suggests that the CYP4 and CYP6 P450 families contain the most highly expressed isoforms in adult C. capitata. Phylogenetic analyses of the conceptual amino acid translations of PCR-amplified cDNAs provides evidence that one of isolated sequences may represent a new P450 family. PMID- 10380099 TI - Voltage-gated sodium channel genes hscp and hDSC1 of Heliothis virescens F. genomic organization. AB - We report the genomic sequence of hscp, a sodium channel alpha subunit gene for Heliothis virescens. A 32-kb genomic clone and six independent RT-PCR products covering almost the entire coding region of the gene, contained thirty-one deduced exons with a translation of 1695 residues. Overall amino acid similarity to the para locus of Drosophila melanogaster was 86%. The transcription of the gene was complex. Alternate splicing was evident for five optional exons and a pair of mutually exclusive exons. A number of alternatively spliced mRNA revealed a deduced translation product that included only the first homology domain. We also report the first partial sequence for hDSC1, a presumed orthologous of the DSC1 sodium channel alpha subunit gene of D. melanogaster. PMID- 10380100 TI - A new aminopeptidase from diamondback moth provides evidence for a gene duplication event in Lepidoptera. AB - We screened a midgut cDNA library from diamondback moth, Plutella xylostella, with a probe generated using sequence information from an aminopeptidase N gene from Manduca sexta (MsAPN-1). The sequence recovered (PxAPN-A) encodes a protein of 988 resides with a 60% sequence identity to MsAPN-1. The two proteins share a signal peptide which directs processing by the endoplasmic reticulum, a C terminal hydrophobic region satisfying the criterion for a GPI anchor and cleavage, and the possibility of an O-glycosylated rigid stalk attached to the GPI anchor. PxAPN-A is more closely related to MsAPN-1 than it is to another aminopeptidase recently reported from P. xylostella. Sequence comparisons with other species suggests that at least one aminopeptidase gene duplication occurred in an ancestral lepidopteran. PMID- 10380101 TI - Typing of sandflies from Greece and Cyprus by DNA polymorphism of 18S rRNA gene. AB - A simple and reliable technique was developed to distinguish Phlebotomine sandflies by restriction fragment length polymorphism of PCR-amplified (PCR-RFLP) 18S rDNAs. Seven morphologically identified sandflies species from several localities of Greece and Cyprus were studied, and specific patterns were developed by double digesting amplified 18S rDNAs with HpaII and RsaI. Three additional species of the subgenus Larroussius were distinguished by a second double digestion with AccI and BanI. We have successfully applied the method on samples in which morphological characters were badly distinguished due to poor storage conditions and in larval stages. PMID- 10380102 TI - Distribution and phylogeny of Wolbachia inducing thelytoky in Rhoditini and 'Aylacini' (Hymenoptera: Cynipidae). AB - Wolbachia are endosymbiotic bacteria responsible for thelytoky in several parasitoid hymenopteran genera. After finding these micro-organisms in some populations of Diplolepis spinosissimae (Hymenoptera: Cynipidae) where they are responsible for thelytoky through gamete duplication, we searched for Wolbachia spp. using specific PCR primers in nineteen other species of the Rhoditini tribe (rose gallwasps) and eight species of the 'Aylacini' tribe (gallwasps associated with herbaceous plants). Wolbachia were found in twelve Rhoditini species and four 'Aylacini' species. The most infected species have very few males (spanandry) and the thelytoky of infected species/arrhenotoky of uninfected species is confirmed by previous research based on the sex of the offspring of virgin females. Phylogenetic analyses based on the partial Wolbachia ftsZ gene sequences indicate that some strains associated with closely related gallwasps are phylogenetically distant, suggesting that cynipids have been affected by several infection events. In contrast, the five infected European species of Diplolepis harbour the same strain of Wolbachia. PMID- 10380103 TI - Structural polymorphism of the luciferase gene in the firefly, Luciola lateralis. AB - To study the structural features of genes for luciferase in Luciola lateralis, two different, functional luciferase genes, Luc1 and Luc2, were cloned and their nucleotide sequences were determined. The two genes were nearly identical, but a deletion of 128 bp was found in the 5' upstream region of Luc2, and minor deletions and additions were also found in the 3' downstream region. Seven base substitutions in the coding region produced two amino acid residue differences between the two genes. Southern blot analysis of genomic DNA isolated from 33 individual fireflies revealed that 26 were homozygous for the Luc1 gene and one was homozygous for Luc2. The others were heterozygotes with two, or even three, different alleles for the luciferase gene. PMID- 10380104 TI - Moose, a new family of LTR-retrotransposons in the mosquito Anopheles gambiae. AB - A novel LTR retrotransposable element called moose has been cloned and characterized from the malaria transmitting mosquito, Anopheles gambiae. This element has all the characteristic features of LTR retroelements and is related to retroelements from other insects and nematodes, belonging to a subgroup of retroelements distinct from the copia/Ty1 and gypsy/Ty3 groups. The moose element appears to be active in A. gambiae, and strong RNA expression is detected in the male and female gonads. The use of this retroelement as a potential vector for germ line transformation is discussed. PMID- 10380105 TI - Genetic similarity among pheromone and voltinism races of Ostrinia nubilalis (Hubner) (Lepidoptera: Crambidae). AB - The genetic variability of seven European corn borer populations, Ostrinia nubilalis, from North America and Europe was assessed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis and DNA sequencing. The nuclear ribosomal internal transcribed spacer 1 (ITS-1) region (approximately 500 base pair [bp]) and four mitochondrial (mtDNA) regions (1550 bp total) were examined. The smartweed borer, Ostrinia obumbratalis, and south Western corn borer, Diatraea grandiosella, were used for comparisons. Of 106 restriction sites identified (80 in mtDNA and 26 in ITS-1), none differentiated geographical populations, pheromone races, or voltine ecotypes of the European corn borer. The lack of variation in the ITS-1 of European corn borer was confirmed by DNA sequence analysis. The genetic similarity of European corn borer populations, despite their wide geographical range and physiological differences, may be explained by a relatively recent origin for the voltinism and pheromone races, gene flow among races, and/or expansion from genetic bottlenecks. PMID- 10380106 TI - Proteins from nuclear extracts of two lepidopteran cell lines recognize the ends of TTAA-specific transposons piggyBac and tagalong. AB - The transposons piggyBac and tagalong are Lepidopteran transposons that exhibit extreme site-specificity for the tetranucleotide TTAA upon insertion and excise in a characteristic precise fashion, regenerating a single TTAA target site. The precise excision of both piggyBac and tagalong can occur in the absence of factors encoded by either transposon, possibly through the recruitment of host proteins and/or cross-mobilizing transposons. In this report, we utilize mobility shift assays and exonuclease III protection analyses to identify DNA binding activities from IPLB-SF21AE and TN-368 cells that are specific for piggyBac and tagalong-terminal repeats. PMID- 10380107 TI - Identification of a non-LTR retrotransposon from the gypsy moth. AB - A family of highly repetitive elements, named LDT1, has been identified in the gypsy moth, Lymantria dispar. The complete element is 5.4 kb in length and lacks long-terminal repeats. The element contains two open reading frames with a significant amino acid sequence similarity to several non-LTR retrotransposons. The first open reading frame contains a region that potentially encodes a polypeptide similar to DNA-binding GAG-like proteins. The second encodes a polypeptide resembling both endonuclease and reverse transcriptase sequences. All members of the LDT1 element family sequenced thus far have poly-A tails or A-rich tails of 12-18 nucleotides in length, but lack a poly-A addition signal in the expected location. The amplification of retrotransposon insertion junction regions in different gypsy moth individuals indicates that polymorphisms exist at some of the insertion sites, suggesting that this element is or was, until recently, capable of transposition. PMID- 10380108 TI - Microbe-induced cytoplasmic incompatibility as a mechanism for introducing transgenes into arthropod populations. AB - Many arthropods are infected with maternally transmitted, intracellular bacteria of the genus Wolbachia. These infections often produce 'cytoplasmic incompatibility' (CI)--reduced egg-hatch frequencies when uninfected females mate with infected males or when males and females carrying different Wolbachia strains mate. Because infected females often enjoy a fitness advantage--they are effectively immune to any effects from males carrying the same Wolbachia strain- Wolbachia and associated cytoplasmic elements can spread rapidly through natural populations. Wolbachia might therefore drive transgenes associated with disease control or pest abatement into populations. In this paper, simple mathematical analyses are presented of three alternative strategies for 'CI drive'. The analyses reveal which parameters must be estimated in order to predict population dynamics, and they demonstrate stringent requirements for initially driving and/or maintaining transgenes in target populations. PMID- 10380109 TI - Cloning and expression of ecto 5-nucleotidase from the cattle tick Boophilus microplus. AB - Although 5'-nucleotidases are ubiquitous in higher vertebrates, the arthropod enzymes have been little studied. The cDNA sequence of the mature 5'-nucleotidase from the tick Boophilus microplus was therefore determined (GENBANK accession number: U80634). The enzyme has 39-41% sequence identity with the vertebrate 5' nucleotidases and contains binuclear metal ion binding sites. There are no significant introns within the coding region of the genomic sequence. Southern blot analysis indicates the presence of multiple related genes encoding 5' nucleotidases. Recombinant tick 5'-nucleotidase was expressed in both Escherichia coli and in baculovirus-infected insect cells. The E. coli recombinant protein was truncated, inactive and produced in abundance. The enzyme was expressed in baculovirus-infected insect cells as a secreted, soluble, glycosylated and enzymatically active protein. This represents the first successful expression and characterization of enzymatically active recombinant 5'-nucleotidase from any organism. Supplementation of the culture medium with 25 microM zinc resulted in a twofold increase in the activity of the expressed protein. The enzyme was purified to homogeneity. It exists under non-denaturing conditions as a homodimer, with an apparent molecular mass of 135 kDa. The Km for the hydrolysis of AMP was 0.37 microM and the k(cat) = 11.5/s, in agreement with data for the native enzyme. PMID- 10380110 TI - Variation in the salivary peptide, maxadilan, from species in the Lutzomyia longipalpis complex. AB - Maxadilan is an approximately 7kDa peptide that occurs in the saliva of the sand fly Lutzomyia longipalpis. This peptide is a potent vasodilator and may also have immunomodulatory effects related to the pathogenesis of leishmanial infections. Variation in the primary DNA and inferred amino acid sequence of maxadilan is reported. Differences were found within and among natural field populations as well as among sibling species. Extensive amino acid sequence differentiation, up to 23%, was observed among maxadilan from different populations. This is a remarkable degree of polymorphism considering the small size of this peptide. The vasodilatory activity of maxadilan was equivalent among recombinant maxadilan variants. All maxadilan variants induce interleukin-6. Predicted secondary structure and hydrophobicity plots suggest that these characteristics are conserved among variant peptides. However, profiles based on the antigenic index do differ among peptides. PMID- 10380111 TI - Identification and characterization of a putative sevenless homologue in the malaria vector Anopheles gambiae. AB - Tyrosine kinase sequences were identified and characterized in Anopheles gambiae, the major vector of malaria in subsaharan Africa. One of these sequences has the characteristics expected for a homologue of the Drosophila sevenless gene, which is necessary for R7 photoreceptor cell fate determination in the developing compound eye. The putative Anopheles sevenless gene homologue is located in a telomeric region of the X chromosome and is expressed in the head of late larval and pupal stage mosquitoes. Identification of the Anopheles homologue of the sevenless gene is a first step towards the development of a dominant phenotypic marker that could be used for detecting transformed Anopheles mosquitoes in a wide variety of genetic backgrounds and, as such, could be used in the development of transgenic mosquitoes for the control of parasite transmission. Preliminary evidence for sevenless sequences were also found in DNA from blackfly, Mediterranean fruit fly and the honeybee. PMID- 10380112 TI - Analysis of genetic variability in Anopheles arabiensis and Anopheles gambiae using microsatellite loci. AB - We analysed genetic variability in Anopheles arabiensis and Anopheles gambiae populations using microsatellite loci to determine whether the Rift Valley restricts the flow of genes. Deviations from Hardy-Weinberg expectations were significant, and were most likely to be due to the high frequency of null alleles observed. An. arabiensis populations occurring between 40 and 700 km apart across the Eastern arm of the Rift Valley were not differentiated (pair-wise F(ST) range: 0.0033-0.0265, P > 0.05). Neither were An. gambiae populations from Asembo Bay and Ghana (F(ST): 0.0063, P > 0.05) despite a geographical separation of about 5000 km. In contrast, significant differentiation was observed between An. gambiae populations from Asembo Bay and Kilifi (about 700 km apart; F(ST) = 0.1249, P < 0.01), suggesting the presence of a barrier to gene flow. PMID- 10380113 TI - Cloning and sequence of a cDNA for a highly basic protease from the digestive juice of the silkworm, Bombyx mori. AB - A serine protease of the silkworm, Bombyx mori, with an isoelectric point of pH 10-11 and a pH optimum for succinyl-Leu-Leu-Val-Tyr-MCA degrading activity of about 10, was found in a 0.33 M NaCl-eluted fraction obtained from cation exchange chromatography of digestive juice. The activity of the enzyme was strongly inhibited by chymostatin and PMSF, indicating that the protease is a chymotrypsin-like serine protease. The N-terminal amino acid sequence of the protease was determined, and a full-length cDNA clone (0.92 kbp) which was isolated from a midgut cDNA library was sequenced. The cDNA encodes a pre proenzyme of 284 amino acids with a pro-segment of 50 amino acids and mature protein of 234 amino acids. From its primary structure, the predicted molecular mass of the mature protein is 24.5 kDa. A sequence comparison of the Bombyx highly basic protease with other serine proteases revealed that this enzyme is a mammalian-type serine protease with a catalytic triad consisting of His45, Asp92 and Ser186. A large number of Arg residues are encoded by the cDNA which may be responsible for its stability and/or function in the alkaline condition, by remaining charged at high pH. PMID- 10380115 TI - Screening of lipase inhibitors from marine algae. AB - The possible presence of an inhibitor of pancreatic lipase (triacylglycerol acylhydrolase, EC 3.1.1.3) was screened in 54 marine algae. An active inhibitor, caulerpenyne, was purified from an extract of Caulerpa taxifolia, using ethyl acetate extraction, followed by successive chromatographies on ODS and silica gel columns. The purified inhibitor was identified by thin-layer chromatography, infrared and nuclear magnetic resonance spectroscopy. Caulerpenyne competitively inhibited lipase activities using emulsified triolein and dispersed 4 methylumbelliferyl oleate (4-MU oleate) as substrates. The concentrations producing 50% inhibition against triolein and 4-MU oleate hydrolysis were 2 mM and 13 microM, respectively. In vivo, oral administration of corn oil with or without caulerpenyne to rats demonstrated a reduced and delayed peak plasma triacylglycerol concentration with caulerpenyne. PMID- 10380114 TI - Fatty acid metabolism in marine fish: low activity of fatty acyl delta5 desaturation in gilthead sea bream (Sparus aurata) cells. AB - Marine fish have an absolute dietary requirement for C20 and C22 highly unsaturated fatty acids. Previous studies using cultured cell lines indicated that underlying this requirement in marine fish was either a deficiency in fatty acyl delta5 desaturase or C18-20 elongase activity. Recent research in turbot cells found low C18-20 elongase but high delta5 desaturase activity. In the present study, the fatty acid desaturase/elongase pathway was investigated in a cell line (SAF-1) from another carnivorous marine fish, sea bream. The metabolic conversions of a range of radiolabeled polyunsaturated fatty acids that comprised the direct substrates for delta6 desaturase ([1-14C]18:2n-6 and [1-14C]18:3n-3), C18-20 elongase ([U-14C]18:4n-3), delta5 desaturase ([1-14C]20:3n-6 and [U 14C]20:4n-3), and C20-22 elongase ([1-14C]20:4n-6 and [1-14C]20:5n-3) were utilized. The results showed that fatty acyl delta6 desaturase in SAF-1 cells was highly active and that C18-20 elongase and C20-22 elongase activities were substantial. A deficiency in the desaturation/elongation pathway was clearly identified at the level of the fatty acyl delta5 desaturase, which was very low, particularly with 20:4n-3 as substrate. In comparison, the apparent activities of delta6 desaturase, C18-20 elongase, and C20-22 elongase were approximately 94-, 27-, and 16-fold greater than that for delta5 desaturase toward their respective n-3 polyunsaturated fatty acid substrates. The evidence obtained in the SAF-1 cell line is consistent with the dietary requirement for C20 and C22 highly unsaturated fatty acids in the marine fish the sea bream, being primarily due to a deficiency in fatty acid delta5 desaturase activity. PMID- 10380116 TI - 3-Thia fatty acid treatment, in contrast to eicosapentaenoic acid and starvation, induces gene expression of carnitine palmitoyltransferase-II in rat liver. AB - The aim of the present study was to investigate the hepatic regulation and beta oxidation of long-chain fatty acids in peroxisomes and mitochondria, after 3-thia tetradecylthioacetic acid (C14-S-acetic acid) treatment. When palmitoyl-CoA and palmitoyl-L-carnitine were used as substrates, hepatic formation of acid-soluble products was significantly increased in C14-S-acetic acid treated rats. Administration of C14-S-acetic acid resulted in increased enzyme activity and mRNA levels of hepatic mitochondrial carnitine palmitoyltransferase (CPT)-II. CPT II activity correlated with both palmitoyl-CoA and palmitoyl-L-carnitine oxidation in rats treated with different chain-length 3-thia fatty acids. CPT-I activity and mRNA levels were, however, marginally affected. The hepatic CPT-II activity was mainly localized in the mitochondrial fraction, whereas the CPT-I activity was enriched in the mitochondrial, peroxisomal, and microsomal fractions. In C14-S-acetic acid-treated rats, the specific activity of peroxisomal and microsomal CPT-I increased, whereas the mitochondrial activity tended to decrease. C14-S-Acetyl-CoA inhibited CPT-I activity in vitro. The sensitivity of CPT-I to malonyl-CoA was unchanged, and the hepatic malonyl-CoA concentration increased after C14-S-acetic acid treatment. The mRNA levels of acetyl-CoA carboxylase increased. In hepatocytes cultured from palmitic acid- and C14-S-acetic acid-treated rats, the CPT-I inhibitor etomoxir inhibited the formation of acid-soluble products 91 and 21%, respectively. In contrast to 3 thia fatty acid treatment, eicosapentaenoic acid treatment and starvation increased the mitochondrial CPT-I activity and reduced its malonyl-CoA sensitivity. Palmitoyl-L-carnitine oxidation and CPT-II activity were, however, unchanged after either EPA treatment or starvation. The results from this study open the possibility that the rate control of mitochondrial beta-oxidation under mitochondrion and peroxisome proliferation is distributed between an enzyme or enzymes of the pathway beyond the CPT-I site after 3-thia fatty acid treatment. It is suggested that fatty acids are partly oxidized in the peroxisomes before entering the mitochondria as acylcarnitines for further oxidation. PMID- 10380118 TI - Effects of dietary supplementation of saturated fatty acids and of n-6 or n-3 polyunsaturated fatty acids on plasma and red blood cell membrane phospholipids and deformability in weanling guinea pigs. AB - The fatty acid composition of plasma cholesteryl esters, plasma phospholipids, red blood cell (RBC) membrane phosphatidylcholine (corresponding to the outer membrane leaflet), and phosphatidylethanolamine (corresponding to the inner membrane leaflet) was investigated in weanling guinea pigs fed with diets of cacao (saturated fatty acids), sunflower oil [n-6 polyunsaturated fatty acids (PUFA)] or fish oil (n-3 PUFA) for 20 wk. RBC deformation was measured by means of a cell-transit analyzer (filtration) and a cone-plate rheoscope. The contents of saturated fatty acids in plasma phospholipids and RBC membrane leaflets were similar in all three groups. Diets with sunflower oil resulted in a high content of linoleic acid in plasma cholesteryl esters and in the outer leaflet of RBC membranes. Fatty acids of fish oil were mainly incorporated in plasma phospholipids and in the inner leaflet of RBC membranes. The arachidonic acid content was high in all groups in the plasma phospholipids and in the inner leaflet. The n-6 and n-3 PUFA were mainly incorporated in the inner leaflet. In all groups the polyunsaturated/saturated fatty acid ratio and the total PUFA content were similar in the inner RBC membrane. The RBC filtration times and the RBC deformation indices were not affected by the dietary treatment. PMID- 10380117 TI - Cross-influence of membrane polyunsaturated fatty acids and hypoxia-reoxygenation on alpha- and beta-adrenergic function of rat cardiomyocytes. AB - The purpose of the present investigation was to determine whether the beneficial effects of polyunsaturated fatty acids (PUFA) may influence ischemia-reperfusion induced alterations of myocardial alpha- and beta-adrenoceptor (alpha-AR, beta AR) responsiveness. This study was carried out using monolayer cultures of neonatal rat ventricular myocytes in a substrate-free, hypoxia-reoxygenation model of ischemia. The cardiomyocytes (CM) were incubated during 4 days in media enriched either with n-6 PUFA (arachidonic acid, AA) or with n-3 PUFA (eicosapentaenoic acid, EPA, and docosahexaenoic acid, DHA). The n-6/n-3 ratio in n-3 CM was close to 1.2, compared to 20.1 in n-6 CM. The contractile parameters of n-6 CM and n-3 CM were similar in basal conditions as well as during hypoxia and reoxygenation. In basal conditions, the phospholipid (PL) enrichment with long chain n-3 PUFA resulted in an increased chronotropic response to isoproterenol (ISO) and to phenylephrine (PHE). After posthypoxic reoxygenation, the chronotropic response to beta-AR activation in n-6 CM was significantly enhanced as compared with the control response in normoxia. In opposition, the ISO-induced rise in frequency in n-3 CM in control normoxia and after reoxygenation was similar. In these n-3 CM, the changes in contractile parameters, which accompanied the chronotropic response, were also similar in reoxygenation and in normoxic periods, although the rise in shortening velocity was slightly increased after reoxygenation. In response to PHE addition, only the chronotropic effect of n-6 CM appeared significantly enhanced after hypoxic treatment. These results suggested that increasing n-3 PUFA in PL reduced the increase in alpha- and beta-AR functional responses observed after hypoxia reoxygenation. This effect may partly account for the assumed cardiac protective effect of n-3 PUFA, through the attenuation of the functional response to catecholamines in the ischemic myocardium. PMID- 10380119 TI - The effects of dietary alpha-linolenic acid compared with docosahexaenoic acid on brain, retina, liver, and heart in the guinea pig. AB - The aim of this study was to compare two different strategies to elevate brain, retina, liver, and heart docosahexaenoic acid (DHA) levels in guinea pigs. First, we used an increasing dose of alpha-linolenic acid (ALA) relative to a constant linoleic acid (LA) intake, and second, we used two levels of dietary DHA provided in conjunction with dietary arachidonic acid (AA). The percentage DHA and AA of total phospholipids in retina, liver, and heart, and in the brain phosphatidylethanolamine and phosphatidylcholine was studied in female pigmented guinea pigs (3 wk old) fed one of five semisynthetic diets containing 10% (w/w) lipid for 12 wk. The LA content in the diets was constant (17% of total fatty acids), with the ALA content varying from 0.05% (diet SFO), to 1% (diet Mix), and to 7% (diet CNO). Two other diets (LCP1 and LCP3) had a constant LA/ALA ratio (17.5:1) but varied in the levels of dietary AA and DHA supplementation. Diet LCP1 was structured to closely replicate the principal long chain polyunsaturated fatty acids (PUFA) found in human breast milk and contained 0.9% AA and 0.6% DHA (% of total fatty acids) whereas diet LCP3 contained 2.7% AA and 1.8% DHA. At the end of the study, animals were sacrificed and tissues taken for fatty acid analyses. We found no significant effects of diets on the growth of guinea pigs. Diets containing ALA had profoundly different effects on tissue fatty acid compositions compared with diets which contained the long chain PUFA (DHA and AA). In the retina and brain phospholipids, high-ALA diets or dietary DHA supplementation produced moderate relative increases in DHA levels. There was no change in retinal or brain AA proportions following dietary AA supplementation, even at the highest level. This was in contrast to liver and heart where tissue DHA proportions were low and AA predominated. In these latter tissues, dietary ALA had little effect on tissue DHA proportions although the proportion of AA was slightly depressed at the highest dietary ALA intake, but dietary DHA and AA supplements led to large increases (up to 10-fold) in the proportions of these PUFA. Tissue uptake of dietary AA and DHA appeared maximal for the LCP1 diet (replicate of breast milk) in the heart. There were no significant changes in the plasma levels of 11-dehydrothromboxane B2 (a thromboxane A2 metabolite) for any diet. The data confirm that dietary ALA is less effective than dietary DHA supplementation (on a gram/gram basis) in increasing tissue DHA levels and that tissues vary greatly in their response to exogenous AA and DHA, with the levels of these long chain metabolites being most resistant to change in the retina and brain compared with liver and heart. Dietary DHA markedly increased tissue DHA proportions in both liver and heart, whereas the major effect of dietary AA was in the liver. Future studies of the effects of dietary DHA and AA supplementation should examine a variety of tissues rather than focusing only on neural tissue. PMID- 10380121 TI - Lipophilic aldehydes and related carbonyl compounds in rat and human urine. AB - Rat and human urine samples were analyzed for lipophilic aldehydes and other carbonyl products of lipid peroxidation. The following compounds were identified as their 2,4-dinitrophenyl hydrazones by cochromatography with pure standards using three solvent systems: butanal, butan-2-one, pentan-2-one, hex-2-enal, hexanal, hepta-2,4-dienal, hept-2-enal, octanal, non-2-enal, deca-2,4-dienal, 4 hydroxyhex-2-enal, and 4-hydroxynon-2-enal. In general, fasted rats excreted less of these compounds than fed rats, indicating they were partially of dietary origin or that the endogenous compounds were excreted in a form not susceptible to hydrazone formation. The compounds excreted in human urine were similar to those excreted in rat urine but were present in lower concentrations. Identification of the conjugated forms of the lipophilic aldehydes and related carbonyl compounds excreted in urine may be a source of information about their reactions in vivo. PMID- 10380120 TI - Dietary supplementation with arachidonic and docosahexaenoic acids has no effect on pulmonary surfactant in artificially reared infant rats. AB - Despite the potential use of long chain polyunsaturated fatty acid (LCPUFA) supplementation to promote growth and neural development of the infant, little is known about potential harmful effects of the supplementation. The present study determined whether supplementation with arachidonic acid (AA) and/or docosahexaenoic acid (DHA) in rat milk formula (RMF) affects saturation of pulmonary surfactant phospholipids (PL). Beginning at 7 d of age, infant rats were artificially fed for 10 d with RMF supplemented with AA at 0, 0.5, and 1.0% of total fatty acid, or supplemented with DHA at 0, 0.5, and 1.0%, or cosupplemented with AA and DHA at levels of 0:0, 0.5:0.3, and 1.0:0.6% of the fat blend. Lung tissue PL contained 43 weight percent palmitate (16:0) of total fatty acids in infant rats fed the unsupplemented RMF. The supplementation with AA at both 0.5 and 1.0% decreased the weight percentage of 16:0 and stearate (18:0), indicating a decrease in saturation of PL. The observed decreases were accompanied by increases in AA and linoleic acid (18:2n-6). Surfactant phosphatidylcholine (PC) consisted of 71 weight percent 16:0 in the unsupplemented group, and this highly saturated PC was not altered by the cosupplementation with AA and DHA although there was a slight increase in DHA. Similarly, the cosupplementation did not change fatty acid composition of surfactant PL when compared with the unsupplemented group. The cosupplementation slightly decreased the weight percentage of 16:0 with a proportional increase in 18:0 leading to an unchanged weight percentage of total saturated fatty acids. These results suggest that, unlike lung tissue PL, the composition of saturated fatty acids in surfactant PL, particularly PC, is resistant to change by dietary AA and DHA supplementation. This, together with the unchanged concentration of total fatty acids in surfactant PC, indicates that LCPUFA cosupplementation causes no effect on pulmonary surfactant. PMID- 10380122 TI - Fenofibrate protects lipoproteins from lipid peroxidation: synergistic interaction with alpha-tocopherol. AB - One of the earliest steps of atherosclerotic plaque formation is an increase of circulating apolipoprotein B-containing lipoproteins which, after infiltrating the subendothelial space, undergo oxidative modification. Fenofibrate is an effective cholesterol- and triglyceride-lowering agent which has been shown to be beneficial in the treatment of atherosclerosis. Vitamin E, or alpha-tocopherol, is a powerful antioxidant which has been shown in a variety of studies to prevent lipoprotein peroxidation. The purpose of the present study was to investigate the effect of fenofibrate treatment, either alone or in combination with alpha tocopherol, in reducing the susceptibility of lipoproteins to oxidative modification. Rats fed a normal diet were treated for up to 27 d with fenofibrate, either alone or in combination with equimolar doses of alpha tocopherol. Combined VLDL (very low density lipoproteins) and LDL (low density lipoproteins) isolated after fenofibrate treatment were more resistant to copper mediated oxidation, as assessed by conjugated diene formation. Lag time was prolonged up to 3.2-fold, while the maximal rate of diene production was significantly decreased by up to 2.2-fold. Treatment of rats with alpha tocopherol alone at the selected dose had no significant effect on lag time, while the propagation rate was slightly decreased. Coadministration of fenofibrate with alpha-tocopherol prolonged the lag phase to a greater extent than fenofibrate alone, showing a synergistic interaction between the two compounds. Finally, the combination of fenofibrate and alpha-tocopherol was significantly more effective in modifying lipoprotein oxidation parameters than what was observed with alpha-tocopherol and bezafibrate or gemfibrozil. Thus, in addition to its well-established effects on lipoprotein concentrations and atherogenic parameters, fenofibrate reduces the susceptibility of VLDL and LDL to oxidative modification and exerts its action synergistically with alpha tocopherol. PMID- 10380123 TI - Microemulsion of seal oil markedly enhances the transfer of a hydrophobic radiopharmaceutical into acetylated low density lipoprotein. AB - Four different microemulsions differing in their core lipid component (triolein, canola oil, squalene, or seal oil) and containing 1,3-dihydroxypropan-2-one 1,3 diiopanoate (DPIP), a potential radioimaging probe, were prepared by means of ultrasonication. The DPIP microemulsions were incubated with acetylated human low density lipoprotein (AcLDL) and the amount of DPIP transferred into AcLDL was examined. The amount of DPIP in the microemulsions expressed as DPIP/oil (w/w) was dependent on the core lipid component of the microemulsion in the order of seal oil (0.19+/-0.04, mean +/- standard deviation) > squalene (0.15+/-0.02) > canola oil (0.12+/-0.02) > triolein (0.07+/-0.004). With the exception of canola oil, all microemulsions were effective in enhancing the transfer of DPIP into AcLDL in comparison with commonly used methods, i.e., direct diffusion and detergent solubilization. DPIP in seal oil resulted in the highest amount of DPIP transferred into AcLDL [309.16+/-34.82 vs. 203.19+/-64.51 using squalene and 151.31+/-28.54 using triolein (DPIP molecules per AcLDL particle)]. For the first time, oil from harp seals, was studied as a major core lipid component of formulating pharmaceutical microemulsions. DPIP in seal oil resulted in the highest transfer of DPIP into AcLDL which is likely due to the highest DPIP concentration found in this microemulsion as well as the high fluidity of seal oil. PMID- 10380124 TI - Cellular uptake and retention measurements of alkylphosphocholines in the SK-BR-3 breast cancer and Molt-4 leukemia cell line using capillary gas chromatography. AB - The determination of cellular content of octadecylphosphocholine (D-19391) and hexadecylphosphocholine (HePC, D-18506), two anticancer agents of the alkylphosphocholine group, using capillary gas chromatography is described. The compounds' cytotoxicity was first determined by the MTT [3-(4,5-dimethyl-2 thiazolyl)-2,5-diphenyltetrazolium] assay, being indicative for the concentration used in the uptake and retention measurements. D-19391 was added to the SK-BR-3 breast cancer cell line and HePC to the Molt-4 leukemia cell line in concentrations of, respectively, 18.6 and 15.0 microM, during a 36-h incubation period at 37 degrees C, 5% CO2. HePC uptake in the leukemia cells was followed by a 24-h reversibility test in drug-free medium. Subsequently, sample clean-up was performed on a weak cation-exchange column. For the quantitative analysis, HePC was used as internal standard for the D-19391 measurements and vice versa. Derivatization of the samples with trimethylsilylbromide was followed by capillary gas chromatographic analysis. From these data we conclude that our uptake results are quite similar with those of a previous study of HePC cellular uptake in the more resistant Caco-2T colon cancer cell line. Without having investigated the mechanism that underlies the cellular uptake results obtained, our study points to no direct correlation between the compounds' cellular uptake and their cytotoxic effects. PMID- 10380125 TI - Steryl esters in the elaioplasts of the tapetum in developing Brassica anthers and their recovery on the pollen surface. AB - The tapetum cells in the developing anthers of Brassica napus contained abundant elaioplasts, which had few thylakoid membranes but were packed with globuli of neutral esters. Of the neutral esters, the major ester group possessed mainly 24 methylenecholesterol, 31-norcycloartenol, 24-dehydropollinastanol, and pollinastanol esterified to 18:3 and other unsaturated and saturated fatty-acyl moieties. The minor ester group had a dominant component tentatively identified as 12-dehydrolupeol esterified to mostly 18:0, 16:0, and 20:0 fatty-acyl moieties. The elaioplasts also contained a high proportion (16% w/w of total lipids) of monogalactosyldiacylglycerols (MGDG). This is the first report of plastids having steryl esters as the predominant lipids. We propose that the globuli contain steryl esters and are stabilized by surface MGDG and structural proteins. The tapetosomes, the other abundant lipid-containing organelles in the tapetum, possessed triacylglycerols (TAG) as the predominant lipids. At a late stage of anther development, the minor group of neutral esters and MGDG of the elaioplasts, as well as the TAG of the tapetosomes, were degraded. Steryl esters similar to those of the elaioplasts were recovered from the pollen surface and were the major lipids of the pollen coat. The pollen coat steryl esters and proteins could be extracted with moderately polar or nonpolar solvents. These proteins, which were mostly fragments of oleosins derived from the tapetosomes, had a high proportion of lysine (13 mol %). The possible functions of the steryl esters and the proteins on the pollen surface are discussed. PMID- 10380126 TI - Occurrence of gamma-linolenic acid in compositae: a study of Youngia tenuicaulis seed oil. AB - Seeds of Youngia tenuicaulis and other species from the plant family Compositae (Asteraceae) were studied for their oil content and fatty acid composition. The seed oil of Y. tenuicaulis growing in Mongolia was found to contain 5.6% gamma linolenic acid (18:3delta6cis,9cis,12cis) in addition to common fatty acids. The oil was analyzed using chromatographic [capillary gas-liquid chromatography (GLC), thin-layer chromatography] and spectroscopic (infrared, gas chromatography mass spectrometry) techniques. Seed oil fatty acids of Saussurea amara (containing gamma-linolenic acid) and of Arctium minus (containing 18:3delta3trans,9cis,12cis), as well as delta5cis- and delta5trans-18:3 were used as GLC reference substances. The evolution in this plant family of a large number of different 18:3 acids as well as the corresponding evolution of unusual desaturases should be investigated. On the other hand, the delta6cis-desaturase required for the biosynthesis of gamma-linolenic acid may have evolved independently several times in unrelated families of the plant kingdom. PMID- 10380127 TI - The 22-kDa antigen in optic nerve and retinal diseases. AB - OBJECTIVE: Patients with unexplained visual loss were evaluated for the possibility of immunologic involvement. Antibody reactions were sought that might identify a common indication of retinal hypersensitivity. METHODS: The enzyme linked immunosorbent assay (ELISA) and Western blot analysis were used to identify autoantibody reactions with retina and optic nerve components. Comparisons were made with the autoantibody reaction of normal subjects and patients with recognized forms of retinal decay: macular degeneration, retinitis pigmentosa, diabetic retinopathy, and paraneoplastic retinopathy. RESULTS: Eight patients, one man and seven women, were found to produce an autoantibody reaction with retina and optic nerve, including a novel 22-kDa neuronal antigen present within the retina and optic nerve. One of the eight had retinopathy associated with melanoma (MAR Syndrome). Seven of the eight patients had electroretinogram abnormalities, varying from mild to severe. Six displayed features of optic atrophy. One patient with progressive visual loss had visual function stabilized after immunosuppressive therapy. CONCLUSIONS: In the eight cases described, unexplained visual loss was associated with autoantibody reactions with retina and optic nerve, including a common antibody reaction with a 22-kDa neuronal antigen found in the retina and optic nerve. All the patients had either an abnormal electroretinogram or optic atrophy. Six patients had both. The 22-kDa immunologic marker may not be directly involved in the patient's vision loss, but rather may be related to a nonspecific destruction of retina and optic nerve. However, the marker may be useful in identifying a specific subgroup of patients for further analysis. PMID- 10380128 TI - From cortical plasticity to unawareness of visual field defects. AB - It was long held that, following alterations in sensory input, structural changes in the primary visual cortex take place only in early life, during so-called "critical periods." Recently, however, it has been established that, in adults, cortical maps in the brain are not fixed, and the cortex does not perform stereotyped operations. Instead, neuronal receptive fields in the cortex can reorganize following deactivation or an altered pattern of activation. Plasticity is essential for the normal adjustment of the brain to modifications in the sensory environment, and for improving perceptual skills and sensorimotor performances. It also plays a crucial role in recovery from damage to the visual system. Cortical remapping generates a filling-in of visual field defects. Consequently, it alters the image perceived. Cortical rearrangement following lesions in the visual pathways does not restore function to the destroyed tissue, but it helps to compensate for gaps in perception. In this review article, we focus on effects of plasticity in the adult visual cortex which are of major importance in the daily practice of neuroophthalmology. Cortical reorganization, together with resulting filling-in, affects the early recognition and evaluation of visual field defects. The importance of brain remapping in these matters is still largely underestimated by clinicians. PMID- 10380130 TI - Comparison of 24-2 and 30-2 perimetry in glaucomatous and nonglaucomatous optic neuropathies. AB - OBJECTIVE: To determine whether the 24-2 Humphrey visual field (HVF) (Humphrey, San Leandro, CA) strategy provides information comparable to that provided by the 30-2 strategy in patients with optic nerve disease. METHODS: In part A of the study, an occluder device was designed to cover the additional outer 22 points tested in the 30-2 strategy of 187 HVFs from neuro-ophthalmology patients with nonglaucomatous optic neuropathy and 206 HVFs from patients with glaucoma. This device converted the gray scale and probability plots of the 30-2 HVF to a 24-2 field. Fields were initially read using the occluder and then were read in a masked manner without the occluder and compared. In part B, 15 healthy volunteers performed both 30-2 and 24-2 HVFs. Testing time and global indices were compared. Ninety-five percent of the fields in the neuro-ophthalmology patients, 96% of the fields in patients under observation for suspected glaucoma, 98% of the fields in patients with ocular hypertension, and 100% of the fields in patients with glaucoma were read similarly with the 24-2 and 30-2 strategies. In the few cases in which a discrepancy was noted between the 24-2 and the 30-2 fields, appropriate clinical management would not have been compromised by using the 24-2 strategy. Most of these cases were in patients with idiopathic intracranial hypertension and very subtle nerve fiber bundle defects. The 24-2 strategy had a significantly lower pattern standard deviation (P < 0.01) and corrected pattern standard deviation (P = 0.05) than did the 30-2 strategy. In addition, the 24-2 strategy shortened the standard threshold testing time by 28% in normal volunteers (P < 0.0001 ). CONCLUSIONS: In most cases, the 24-2 testing strategy provides information comparable to that provided by the 30-2 strategy in a shorter time and with less variability. A 30-2 HVF may be warranted in patients under observation for evolving idiopathic intracranial hypertension. PMID- 10380129 TI - Comparison of threshold visual perimetry and objective pupil perimetry in clinical patients. AB - OBJECTIVES: In an attempt to measure the visual field objectively, we have performed pupil perimetry, by which the pupil light reflex is monitored in response to perimetric light stimuli. The purpose of this study was to ascertain whether pupil perimetry reveals defects similar to those revealed by standard threshold perimetry in patients with various diseases. MATERIAL AND METHODS: An infrared pupillometer was linked to an automated perimeter to record, at each perimetric location, 76 pupil contractions, which were comparable to the test locations of the Humphrey Field Analyzer (HFA 30-2 program; Humphrey, San Leandro, CA). One hundred eighteen patients with various diseases were investigated. RESULTS: Ninety-one patients (77.1%) maintained a pupil area large enough (more than 10 mm2 in area) to respond adequately to focal light stimuli throughout the test. The correlation between the pupil field and the threshold visual field was subjectively judged to be good in most cases. However, pupil perimetry showed less damage than that seen in threshold perimetry in six of nine patients who had Leber's hereditary optic neuropathy (LHON). CONCLUSIONS: Pupil perimetry is a good method for measuring the visual field objectively and has potential for clinical use in most of the cases. PMID- 10380131 TI - Bilateral trochlear nerve palsy associated with cryptococcal meningitis in human immunodeficiency virus infection. AB - This is the report of a case of bilateral trochlear nerve palsy secondary to cryptococcal meningitis in a 34-year-old woman with acquired immune deficiency syndrome. Based on clinical and neuroradiologic findings, it is concluded that in the present case, a postinflammatory shrinking of the arachnoid has stretched the fourth cranial nerves at their point of emergence from the dorsal surface of the brainstem. PMID- 10380132 TI - Three patients with presumed rhino-orbital-cerebral mucormycosis. PMID- 10380133 TI - Spontaneous resolution of upper eyelid retraction in thyroid orbitopathy. AB - This study was conducted to document in the literature case reports of spontaneous resolution of eyelid retraction in patients with thyroid orbitopathy. Two cases of thyroid orbitopathy associated with eyelid retraction were observed without surgical treatment. Spontaneous resolution of upper eyelid retraction occurred during an 8- to 12-month period. PMID- 10380134 TI - A case of ocular neuromyotonia with tonic pupil. AB - A 48-year old woman with hypertension experienced painful oculomotor nerve palsy. After surgery for a giant aneurysm of the internal carotid artery in the cavernous sinus, phasic constrictions of the pupil developed. Two years later, this phenomenon disappeared and was replaced by intermittent involuntary cyclic spasms elevating the ptosed lid. These cyclic lid movements were not elicited with any eye movement or by increased accommodation. The pupil now manifested the pharmacologic features of a tonic pupil. The explanation for this unique case of ocular neuromyotonia is based on a misdirection phenomenon, possibly caused by ephaptic transmission. PMID- 10380135 TI - The perils of a sneeze. AB - A 51-year-old woman had a 3-day history of severe left supraorbital pain associated with blurred vision of the left eye. Examination revealed visual acuity of 20/20 OD and 20/100 OS. A left relative afferent pupillary defect was present. Neuroimaging revealed a large intra-, supra-, and parasellar mass that had eroded through the sphenoid sinus into the maxillary sinus. Secondary pneumocephalus was present. Pathologic examination of the tissue revealed a pituitary adenoma of the null cell type. To the best of our knowledge, there is only one other case in the literature in which a spontaneous pneumatocele represents the initial manifestation of a pituitary adenoma. PMID- 10380136 TI - Wall-eyed bilateral internuclear ophthalmoplegia in central nervous system cryptococcosis. AB - Only one case of wall-eyed bilateral internuclear ophthalmoplegia (WEBINO) has been described in central nervous system cryptococcosis. The disorder was initially unilateral, then became bilateral with skew deviation and vertical upgaze deficit. We report a case of WEBINO in central nervous system cryptococcosis in a patient with acquired immune deficiency syndrome. Magnetic resonance imaging revealed high signal on T2 images in the right midbrain, left frontal vertex, left splenium, and cerebellum. With treatment, the internuclear ophthalmoplegia improved; however, the convergence insufficiency remained. Disruption of input from cortical supranuclear locations or the region of the rostral interstitial nucleus of the medial longitudinal fasciculus has been proposed as a mechanism in the absence of convergence. This correlates in our patient with the lesions seen on magnetic resonance images. PMID- 10380137 TI - Hemianopsia related to dissection of the internal carotid artery. AB - Spontaneous dissection of the internal carotid artery is typically associated with cerebral vascular infarction along the anterior and middle cerebral distribution, whereas occipital infarction is usually related to posterior circulation abnormalities. Hemianopsia with occipital infarction related to carotid artery dissection has therefore rarely been reported. A 40-year-old woman in whom acute-onset hemianopsia developed, related to occipital infarction secondary to internal artery dissection, is described. This atypical association is explained by anatomic variations of the posterior part of the circle of Willis. Neuroimages showed occipital infarction related to internal carotid artery dissection associated with hypoplasia of the proximal portion of the cerebral posterior artery (P1). The anatomic correlation of this atypical association and a review of the literature are presented. PMID- 10380138 TI - Disseminated histoplasmosis causing reversible gaze palsy and optic neuropathy. AB - Subacute disseminated histoplasmosis is an uncommon entity. Typical neuro ophthalmologic manifestations are usually secondary to histoplasmomas or encephalitis. A 45-year-old man noted blurred vision while receiving empiric antituberculosis therapy for fever and diffuse granulomatous disease of unknown origin. Vertical-gaze palsy, right horizontal-gaze paresis, and mild right optic neuropathy were found on neuro-ophthalmologic examination. Further questioning revealed a history of frequent contact with fighting cocks from South America. Magnetic resonance images were consistent with multiple hemorrhagic infarcts, areas of inflammation, or both, and cerebral angiography showed changes consistent with vasculitis. A previously obtained biopsy specimen from the duodenum was restained and found to be positive for fungal elements. Serum antigen titers for Histoplasma capsulatum demonstrated evidence of active infection. This case is a rare example of a supranuclear ocular motility disturbance and optic neuropathy secondary to an occlusive vascular process in a patient with subacute disseminated histoplasmosis. PMID- 10380139 TI - Occurrence of familial nonarteritic anterior ischemic optic neuropathy in a case series. AB - OBJECTIVE: To report on the occurrence of familial nonarteritic anterior ischemic optic neuropathy (NAION) in our NAION series. METHODS: One hundred forty-eight consecutive retrievable cases of NAION were surveyed regarding the occurrence of NAION in other family members. Medical records of affected family members were reviewed, and clinical characteristics of documented familial NAION cases were described. RESULTS: Of 79 patients who returned the survey, four reported one or more relatives with previously diagnosed NAION. There were nine cases of documented NAION in these four families. All cases occurred in siblings, with a mean age at onset of 55 years. Six patients had second eye involvement and in five, involvement became bilateral within 4 years after initial onset. None of the patients had diabetes; two had hypertension. CONCLUSION: A small number of patients with NAION may belong to a familial subclass. Three previous reports of familial NAION further support this hypothesis. PMID- 10380140 TI - Traumatic Horner syndrome without anhidrosis. AB - In a patient with a traumatic avulsion of the arm, magnetic resonance imaging showed the exact site of the lesion that produced Horner syndrome with preservation of sweating on the face. PMID- 10380142 TI - [Seville Quality of Life Questionnaire: historic outlook of its establishment]. AB - We describe how we developed a new instrument for measuring quality of life, the Seville Quality of Life Questionnaire (SQCQ), and the results we obtained when we applied it to a group of schizophrenic patients. The questionnaire was designed by a group of researchers who set out from the premise that the quality of life of schizophrenic patients is perceived differently from that of healthy people, and tried first of all to demarcate the kind of area which is abnormal in these patients. We then devised a set of items which were assessed for clarity and pertinence by a series of experts, and the questionnaire was assembled in the form of a set of statements, response to which is in the form of a Lickert-type 5 stage scale. The finished questionnaire was administered to 279 schizophrenic patients. At the same time, these patients were also evaluated using the AMDP Psychopathology Scale, the positive and negative symptoms evaluation scale (PANS), Lehman's structured quality of life interview (QOLY), Ruiz and Baca's quality of life questionnaire (QLQ), Camberwell's scale of needs analysis (CANr) and the WHO's handicap assessment scale (DDS). These questionnaires were used to judge the different types of validity of the Seville Questionnaire. The reliability of the questionnaire was measured using Cronbach's alpha coefficient (internal consistency: 0,85 scale of favourable aspects, 0,96 scale of unfavourable aspects). As far as validity was concerned, both scales of the questionnaire were found to have a high level of validity. We also examined the extent to which the psychopathological disorders affected the quality of the schizophrenic person's life, the extent to which his/her needs were being met by the health services, and the patient's degree of disability. From the results obtained, we can say that the SQCQ is a reliable and valid instrument, which is sufficiently sensitive to the clinical changes produced in the course of the natural history of the disease. The SQCQ stands out from the other quality of life questionnaires in that it takes into account aspects of the disease which the patient him/herself is aware of as affecting his/her quality of life. PMID- 10380141 TI - [Long term evolution of the incapacity in schizophrenic patients in maintenance treatment with risperidone]. AB - OBJECTIVE: To determine the evolution of the degree of long term disability (8 months) in a group of schizophrenic outpatients undergoing monotherapy with risperidone. DESIGN, An observational multicentre study of 8 months follow up. PATIENTS: 354 patients with schizophrenic disorder (ICD-10). EVALUATION: Baseline, 2, 4 and 8 months. INSTRUMENTS: BPRS, CGI, UKU, WHO/DAS-S. RESULTS: a significant decrease in both the global scores and in each of the 4 areas of disability. Improvement in disability depends to a large degree on the improvement of the disorder as shown on the BPRS and CGI. After 8 months, those patients with paranoid subtype and the less severe ones (BPRS and CGI) showed a considerably lesser degree of disability. The final level of disability (square root = 0,61) is narrowly related to the baseline level of disability, the 3 clusters of the final BPRS, the final CGI, gender and the subtype of schizophrenia. PMID- 10380143 TI - [The chronic fatigue and neurasthenia in the student population]. AB - INTRODUCTION: Fatigue is one of the most common symptoms in community studies, primary care and other medical setting. In spite of a high frequency of fatigue, the incidence of chronic fatigue syndrome is very low. In this paper, we want to know the frequency of chronic fatigue syndrome and neurasthenia; we want to know the association between fatigue and depressive symptoms in students. METHODS: We studied 277 medical student, administering: 1. a center for disease control questionnaire to assess major criteria and minor criteria of chronic fatigue syndrome, 2. ICD 10 criteria for the diagnoses of neurasthenia and 3. Beck depression inventory. RESULTS AND CONCLUSIONS: We found that the 37,55% of the subjects suffer fatigue. 9 subjects (3,25% of the total) meet the criteria of neurasthenia. 2 subjects (0,72% of the total) meet the chronic fatigue syndrome criteria. The depressive symptoms are most frequent in the subjects with fatigue, but we don't know if they are the cause or the consequence of the fatigue. With the factorial analyses, we find that symptoms of physical fatigue, mental fatigue and cognitive difficulties are factor independent of each other. PMID- 10380144 TI - [Sexual dysfunction with antidepressive agents. Effect of the change to amineptine in patients with sexual dysfunction secondary to SSRI]. AB - Sexual dysfunction secondary to the use of antidepressants, especially clomipramine or SSRI's is an adverse effect that is often underestimated and according to earlier studies, this can affect approximately 60% of the patients. This presents as a decrease in libido, alterations in the ability to reach orgasm/ejaculation, and an erectile dysfunction or a decreased vaginal lubrication. This dysfunction appears to be related with the resulting increase in serotonin and with the stimulation of serotonin 5HT2 receptors. OBJECTIVES: 1) Evaluate the effect of amineptine, a drug with an increased dopamine transmission and scant serotonin transmission, on the sexual function of depressed patients who begin treatment, and 2) evaluate whether the change to amineptine improves the sexual function in patients who presented sexual dysfunction after beginning treatment with a SSRI. MATERIAL AND METHODS: Prospective, observational, open and multicentric design. 111 patients with an average age of 41.3 years (36 men, 75 women) were distributed into three groups: Group 1 (n= 26): patients with depression (DSM IV) who begin de novo treatment with amineptine 200 mg/day. Group 2 (n= 47): depressed patients undergoing treatment with a SSRI who show a favorable response and who present sexual dysfunction secondary to a poorly tolerated treatment, so the treatment is changed to 200 mg/day of amineptine. Group 3 (n= 38): patients with the same characteristics as those of group 2, but whose treatment was changed to 20 mg/day of paroxetine. The <> (Montejo et al, 1996) was used together with the Hamilton Depression Scale, the IGC Scale, and an adverse events scale, over a 6 months follow up period during which visits took place at: baseline, month 1, month 2, month 3, and month 6. RESULTS: In group 1, treated with amineptine from the beginning, of the 5 patients who showed a decrease in the libido at the beginning of the treatment, only one still presented this in the 6th month. The Hamilton Scale decreased from 23.12 (baseline) to 5.25 after 6 months. After substituting amineptine for SSRI's in patients with sexual dysfunction, the incidence of any type of sexual dysfunction decreased significantly from 100% (baseline) to 55.3% after 6 months. (P< 0.001). The incidence of delayed orgasm dropped to 15.8%, anorgasmia to 17.4%, and impotence dropped to 15.8% in this group, with the antidepressant effect that had already been achieved with the SSRI being maintained. However, in group 3 there was barely any improvement on the sexual function after changing to paroxetine (20 mg/day), with the baseline incidence being 100% and the incidence after 6 months being 89.7%. In this last group the antidepressant effect present at the baseline level, was maintained. CONCLUSIONS: Amineptine was shown to be an effective antidepressant in the patients studied, and did not cause secondary sexual dysfunction, and even improved the dysfunction that was present in some patients. In those patients previously treated with SSRI's, amineptine is able to significantly improve the sexual dysfunction and yet maintain the efficacy of the antidepressive treatment used before these 6 months. On the other hand, Paroxetine did not improve the sexual dysfunction of the people in whom this drug substituted another SSRI, as this is an adverse effect common to the entire group of selective serotonin re-uptake inhibiting drugs. Amineptine showed a good safety and tolerance profile. Its most common side effect (anxiety/restlessness) disappeared 2 months after the beginning of the treatment. PMID- 10380145 TI - [Analytical thyroid disturbances in psychiatric inpatients]. AB - BACKGROUND: There is not an agreement about the screening for thyroid disease in psychiatric inpatients. METHODS, The thyroid status of 172 psychiatric inpatients was assessed at the beginning of their hospitalization. A logistic regression was performed lo find the factors related with abnormal levels of thyroid hormones. RESULTS: As many as 30.8% of the inpatients presented levels of thyroid hormones and 5.2% have thyroid disease. The model chi2 was excellent (chi2= 20.89; gl= 4; p< 0.0001), the sensitivity was de 0.38 and the specificity was 0.83. Five variables fulfilled the criteria to be entered and not removed from the model: sex, schizophrenia, previous mental disorder, treatment with lithium, current infectious illness. CONCLUSIONS: A screening for thyroid disorder in psychiatric inpatients, at the beginning of their hospitalization, is recommended in female patients, schizophrenics and patients in treatment with lithium. PMID- 10380146 TI - [The role of the personality in the feeding behavior disorders]. AB - Personality disorders are common in eating disorders and preliminary reports indicate that this type of disorders implicate a poor prognosis in anorexia and bulimia nervosa. Cluster C personality disorders, particularly avoidant and dependent personality disorder are the most frequent in anorexia nervosa. In bulimia nervosa, however, cluster B personality disorders, including borderline and histrionic personality disorders, are more frequent. Furthermore, temperament could differentiate anorectic patients from bulimic patients. Eating disorders seem to be associated with high scores in neuroticism and harm avoidance. However, while anorexia nervosa patients might have higher persistence, bulimia nervosa patients seem to have an impulsive temperament. According to the character model of Cloninger, both anorexia and bulimia present lower scores in the dimension self-directedness. PMID- 10380147 TI - [Transcranial magnetic stimulation in Psychiatry]. AB - Transcranial Magnetic Stimulation (TMS) is an exciting new technology that along with repetitive TMS (rTMS) offers the potential to explore and understand brain behavior relationship in a way that builds on recent advances in functional neuroimaging (ie, PET, SPECT, fMRI imaging). rTMS as a relatively noninvasive probe of cortical function provides an opportunity to explore the relationships between regional brain activity and symptomatology across psychiatry illnesses. In this article we briefly review the current thinking regarding the neurobiology of mood and the effects of rTMS on mood in healthy and depressed subjects. PMID- 10380148 TI - [Anxiety disorders and substance-related disorders]. AB - Psychiatric comorbidity between anxiety disorders and substance-related disorders is studied. This relationship is complex due to the overlapping of symptoms (during the abstinence of alcohol and opiates as well as in the intoxication of cocaine) and also for the inaccurate definition of several disorders. Epidemiologic studies show an increase of prevalence of anxiety disorders, mainly agoraphobia and social phobia, in patients who are alcohol dependents and in heroine and cocaine users. Such studies also point out an augmentation of consume of substances (drugs and alcohol), with a prevalence of 24% in patients with several anxiety disorders. The relationship between these disorders is analysed. It is also studied the influence of such a comorbidity in the evolution and outcome of treatment. PMID- 10380149 TI - [Mixed anxiety and depression disorder: a naturalistic study]. AB - INTRODUCTION: We sought to examine the prevalence, clinical characteristics and one-year outcome of Mixed Anxiety-Depressive Disorder (MADD) in a clinical sample. METHODS: We interviewed 400 psychiatric outpatients, diagnosing MADD with DSM-IV research criteria. RESULTS: Forty-two patients (10.5%) were diagnosed of MADD. Two-thirds were females, 50% had chronic medical conditions, 47.6% reported environmental problems, and the mean GAF was 54.4. At follow-up, MADD proved to be a stable diagnosis. Two subgroups emerged according to illness duration, with a chronic subtype (more than six months' duration) significantly different in the variable 12-month full remission. CONCLUSIONS: MADD, an often disabling disorder, appears to be a frequent and stable diagnosis. We further suggest the existence of a distinct chronic subtype. PMID- 10380150 TI - [The recognition of mental disease in primary health care and its determining factors]. AB - AIMS: It is known that recognition of mental illness by the General Practitioner (GP) is low. The GP usually identify less than a half of these cases. Our aim has been to study the prevalence of mental disorders in Primary Care, and to analyse the influence of several variables over the identification of mental illness by the GP. METHODS: Transversal study in four Primary Care centres in the north of Spain. We studied 823 patients attended with a <> in primary care practices. Patients were evaluated with the GHQ-28, Belloc questionnaire, and data regarding diagnosis and treatment provided by the GP, in addition of additional information from medical records. RESULTS: The prevalence of mental illness according the GHQ-28 was 33%, being higher in women (38%) than in men (24%). Mental illness diagnosed by the GP was the 14,1% of cases. The rate of patients with mental disorders (diagnosed by the GHQ) recognised by the GP was the 29%. In a logistic regression, relevant factors affecting recognition were i) presentation with physical symptoms, ii) clinical severity (measured by GHQ), and, iii) employment (only for older patients). CONCLUSIONS: <> can explain the low rate of identification of mental illness by the GP. PMID- 10380151 TI - [Predictors of behavioural disorders and consultation-liaison psychiatry in patients with HIV disorders]. AB - OBJECTIVE: The aim was to identify the features of patients admitted into an Infectious Disease Unit in a Hospital due to pathology related to HIV and/or addiction that originate the demand for consultation liaison psychiatry. METHOD: During the first six months of 1997, 232 admissions into the Infectious Disease Unit at Ramon y Cajal Hospital (191 patients) were systematically evaluated by the nursery staff. A specific questionnaire was designed for this interview. After having performed univariants analysis, a logistic regression was used to identify the most relevant variables in the claim for consultation liaison. RESULTS: The claim for consultation liaison was associated to: consume in the unit OR (yes/no)= 7.9, confusional syndrome OR (yes/no)= 5.6, social worker consultation liaison OR (yes/no)= 2.1. use of benzodiazepines OR (yes/no)= 2.4. No medical treatment respect to bad accomplishment OR= 3.6. Only taking into account the known features after the first examination: use of benzodiazepines OR (yes/no)= 2.1, use of cocaine OR (yes/no)= 1.8, recognized income (yes/no)= 2.2, no medical treatment due to bad accomplishment OR= 2.1. CONCLUSIONS: In our environment the demand for consultation liaison psychiatry is related to behavioural problems due to the use of substances and marginality. The variables which predict the demand and may be identified at the time of admission are: the use of benzodiazepines, cocaine, the lack of a recognized income and the absence of medical treatment for the HIV infection. Recognizing these features will allow us to identify patients who are going to have behavioural problems and demand psychiatric assistance. PMID- 10380152 TI - [Drug interactions of methadone with CNS-active agents]. AB - Psychoactive medication is frequently used in methadone maintenance treatment programs (MMP) to treat comorbid mental disorders (depression, anxiety, schizophrenia) in opiate-addicts. Thus, several pharmacological interactions are possible. This problem becomes more relevant with the introduction of new CNS drugs like SSRI, atypical antipsychotics or new anticonvulsants. The most common interactions seen in practice are pharmacodynamic in nature, most often due to the cumulative effects of different drugs on the central nervous system (e.g. neuroleptics or benzodiazepine interactions). However important pharmacokinetic interactions may occur particularly between methadone and antidepressant drugs: Desipramine plasma levels are increased by methadone; further fluvoxamine (and fluoxetine to a less extent) may cause an important increase in serum methadone concentrations. The inhibition of different clusters of the cytochrome P450 system are involved in these interactions. Several lines of evidence suggest that benzodiazepines and methadone may have synergistic interactions and that opiate sedation or respiratory depression could be increased. This is a serious problem, given the widespread use of benzodiazepines among MMP patients. Experimental but not clinical data support methadone and lithium interactions. Finally, classic anticonvulsant drugs, such as phenytoin, carbamazepine and phenobarbital, produce dramatic decreases in methadone levels, which may precipitate a withdrawal syndrome; valproic acid and the new anticonvulsant drugs do not have these effects. Accordingly, caution is advised in the clinical use of methadone when other CNS-drugs are administered. PMID- 10380153 TI - [Frontotemporal dementia]. AB - Defined barely one decade ago, Frontotemporal Dementia emerges as a new clinical entity that reestablishes the classical Pick's disease as part of a extensive syndrome with important and unsuspected prevalence. Frontotemporal Dementia includes all primary degenerative processes starting in the anterior portions of the brain. This type of dementia is clinically characterized by behavior and personality disorders, more than cognitive alterations. In the present research we performed a bibliographic review of the subject including clinical characteristics, laboratory tests, and neuropathology. Furthermore we assessed areas that require further development. PMID- 10380154 TI - [Archetypal aspects of aging: from Eros to Thanatos]. AB - From a Junguian point of view, the different archetypal roles of the aging process, along the life cycle, are studied: eros, puer, logos, hero, persona, mascara, anima, animus, king, warrior, magician, lover, mother, virgin, afrodite, witch, self, senex, shadow, tanatos. PMID- 10380155 TI - [Treatment with risperidone of a case of spontaneous orgasm]. AB - A case report is presented of a patient who had spontaneous orgasms following treatment with clomipramine and were successfully treated with risperidone. The relationship with previous reports on drug-induced spontaneous orgasm is analyzed. PMID- 10380156 TI - Lung cancer mortality and diesel exhaust: reanalysis of a retrospective cohort study of U.S. railroad workers. AB - A retrospective cohort study of 55,407 U.S. railroad workers has been called the most definitive study linking exposure to diesel exhaust (DE) with lung cancer in humans. However, reanalysis of data from this study suggests caution in interpreting this study as demonstrating such a link. Although workers who rode trains had a significantly elevated lung cancer mortality relative to clerks and signalmen (who were assumed to be unexposed), shop workers did not, despite convincing evidence that these workers had the highest exposures to DE. Mortality from heart disease and cirrhosis of the liver were also significantly elevated among train riders, which suggests that these workers had a substantially different lifestyle from other workers, and raises the possibility that their elevated lung cancer mortality may be related to lifestyle rather than to DE exposure. Smoking information was not available for this cohort. A positive, monotone dose-response trend in lung cancer mortality with increasing duration of exposure found by the original investigators was not present when age was controlled more carefully and years of exposure quantified more accurately. Instead, a negative dose-response trend for lung cancer was seen among exposed workers based on either duration of exposure or quantitative measures of cumulative exposure. Similar negative trends were seen with several broad categories of mortality, including all causes. These negative trends are possibly a result of incomplete follow-up that was most severe among workers with the longest tenures. A sizable fraction of deaths occurring during the last 4 years of follow-up evidently were not identified, and there is evidence that follow-up in earlier years was also incomplete. At the very least, problems with the follow up should be rectified before any conclusions are drawn about the carcinogenicity of DE in this cohort. PMID- 10380157 TI - Xenobiotic-metabolizing enzymes in the canine respiratory tract. AB - Airway epithelial surface is the primary target of airborne pollutants. To estimate the distribution of xenobiotic-metabolizing enzymes in the respiratory tract of dogs, epithelia from different airway sites of four animals were analyzed for metabolism of sulfite (sulfite oxidase) and formaldehyde (formaldehyde dehydrogenase and aldehyde dehydrogenase). In addition, glutathione S-transferases were assayed using several model substrates. Enzyme activities were compared with those found in liver parenchyma. The activity of sulfite oxidase was found to be comparable in nose, trachea, and proximal and medium bronchi, but appeared to be lower in lung parenchyma of most animals. In contrast, hepatic sulfite oxidase activity of these animals was substantially higher compared to that in airway epithelia. The activity of glutathione dependent formaldehyde dehydrogenase (FDH) appeared to be highest in nose and lowest in distal bronchi, lung, and liver parenchyma. The distribution pattern of the glutathione-independent aldehyde dehydrogenase (AldDH) in the respiratory tract was different from that of FDH. Levels of AldDH were about 5- to 10-fold lower than those of FDH, suggesting that AldDH is of minor importance for pulmonary formaldehyde detoxification. With regard to ethanol detoxification by a class I alcohol dehydrogenase (ADH), no measurable enzyme activity could be detected at most respiratory sites contrary to the high activity found in liver parenchyma. Regarding glutathione S-transferases (GSTs), different distributions of enzyme activities were found in the large and small airways when using three substrates. The 1-chloro-2,4-dinitrobenzene (CDNB)-related activities in the cytosolic fraction of the upper (nose, trachea) and lower airways (proximal, medium and distal bronchi) were higher than those in the microsomal fraction. Interestingly, there was no difference between CDNB-related activities in the cytosolic and microsomal fraction of the liver. Highest cytosolic activities were found in the nose, and were comparable to those detected in the liver parenchyma. The cytosolic 1,2-dichloro-4-nitrobenzene (DCNB)-related activities in the nose, proximal bronchi, and lung parenchyma were appeared to be markedly higher than those in trachea and medium and distal bronchi, while the microsomal activities were not detectable at most respiratory sites. In contrast, distinctly higher activities were measured in both fractions of liver tissue. Cytosolic 1, 2-epoxy 3-(p-nitrophenoxy)-propane (EPNP)-related activities were present in upper and lower airways including lung parenchyma at comparable levels, while in liver tissue the mean activities were distinctly lower. No EPNP-related activities were found in the microsomal fractions. In conclusion, most xenobiotic-metabolizing enzymes investigated in this study could be detected in epithelia of various respiratory sites. The most outstanding result revealed higher levels of FDH activity in the nose and downstream to the medium bronchi in comparison to those found in the small airways, lung, and liver tissue. Similarly, the EPNP-related GST exhibited a distinctly higher activity at all respiratory sites compared to the activity in liver tissue, suggesting a different regulation of this enzyme in lung and liver. PMID- 10380158 TI - Metals associated with both the water-soluble and insoluble fractions of an ambient air pollution particle catalyze an oxidative stress. AB - One potential mechanism of injury mediated by air pollution particles is through metal-catalyzed oxidant generation. In one emission source particle, soluble metals have been associated with biological effect and toxicity. However, a majority of metals in ambient air pollution particles can be associated with insoluble components. We tested the hypothesis that concentrations of catalytically active metal in ambient air pollution particles are not equivalent to the concentrations of water-soluble metal. Twelve filters collected from the North Provo, UT, monitoring station were agitated in deionized water. Both the aqueous extract and pellet were isolated, lyophilized, and defined as the water soluble and insoluble fractions, respectively. The fractions were chemically characterized and ionizable concentrations of metals were measured using inductively coupled plasma emission spectroscopy. While the water-soluble fraction had significantly greater concentrations of ionizable metals per unit mass, the insoluble fraction also had measurable quantities. In vitro oxidant generation by the two fractions, measured as thiobarbituric acid-reactive products of deoxyribose, corresponded to the concentrations of ionizable rather than total metals. The release of interleukin-8 by cultured respiratory epithelial cells after incubation with the two fractions also coincided with the ionizable metal concentrations. Finally, neutrophil influx and lavage protein levels 24 h after instillation of the two fractions in rats reflected the ionizable metal concentrations, in vitro oxidative stress, and mediator release. We conclude that catalytically active metals can be measured in both the soluble and insoluble fractions of an ambient air pollution particle. These metals corresponded to the biological activity of the two fractions. While in greater concentration in the water-soluble fraction, larger total quantities of catalytically and biologically active metals are likely to be associated with the insoluble fraction as a result of the abundance of the latter. PMID- 10380159 TI - Pathological and immunological effects of respirable coal fly ash in male Wistar rats. AB - In this study the effects of inhalatory exposure to coal fly ash on lung pathology and the immune system in rats were examined. Rats were exposed to 0, 10, 30, or 100 mg/m(3) coal fly ash (6 h/day, 5 days/wk) for 4 wk, or to 0 and 100 mg/m(3) for 1 wk, and for 1 wk followed by a recovery in clean air of 3 wk. A concentration-related increase in lung weight was found starting from 30 mg/m(3) coal fly ash. After exposure to 100 mg/m(3), a time-related deposition of free particles in the lungs was observed as well as a time-related number of coal fly ash particles phagocytized in alveolar macrophages. Histological examination revealed increased cellularity in alveolar septa, consisting mainly of mononuclear cell infiltrate, proliferated type II cells, and a slight fibrotic reaction. After a recovery period of 3 wk the histological picture was identical to that after 1 wk of exposure, indicating no significant recovery. No toxicological significant changes were found in the hematological, clinical chemistry, or urine parameters. Effects both on nonspecific defense mechanisms and on specific immune responses were noted. With regard to the immune function in the draining lymph nodes of the lung, a significantly increased number of both T and B lymphocytes was observed. The ratio of both cell types was not changed in either of the groups. In serum of exposed rats a significant increase of up to 150% of the immunoglobulin A (IgA) content was found. The number and phagocytic capacity of macrophages were significantly increased, while the killing of Listeria bacteria per cell ex vivo/in vitro remained unchanged. Natural killer (NK) activity in pulmonary cell suspensions was slightly stimulated in rats exposed for 4 wk to 10 and 30 mg/m(3), whereas an exposure to 100 mg/m(3) resulted in a slight decrease; however, both changes were not significant. In conclusion, the alterations in lung histopathology and immunity, observed in a dose and exposure time relation at concentrations up to and including 100 mg/m(3) coal fly ash, may be considered an adverse response of the host to inhalation of particulate matter. Whether these observed alterations may effect the host resistance must be learned from infection studies. PMID- 10380160 TI - A centrifugal particle concentrator for use in inhalation toxicology. AB - Epidemiologic studies have provided strong evidence that episodic exposure to ambient particulate matter is associated with increases in morbidity and mortality. These adverse effects have been demonstrated at concentrations far below the National Ambient Air Quality Standard (NAAQS), and thus, the biological plausibility of these effects has been questioned. For the purpose of exposing test animals to relevant and reproducible exposure concentrations of ambient particulate matter (PM), we have developed a simple and inexpensive concentrator system that can concentrate ambient particles 10-fold. A high-volume blower is used to deliver ambient air to the inlet manifold of a centrifugal concentrator and the entrained particles travel along a concentric annulus formed by a stationary solid outer cylinder and a porous inner cylinder rotating at high speed (up to 12,500 rpm). Suction applied at one end of the porous shaft causes the dispersion medium (air) to pass through the porous cylinder and into the shaft. Since the rotational velocity of airborne particles is comparable to that of the rotating cylinder near its surface, the particles move radially outward due to the centrifugal force, in addition to their motion laterally along the cylinder and inward due to the suction of air into the rotating porous cylinder. The particles reach their highest concentration near the outlet manifold, where they enter the exposure chamber under positive pressure ( approximately 0.4 cm H2O). Except for coarse particle loss due to impaction and diffusional loss of ultrafine particles in the concentrator, the increase in particle concentration is the ratio of the flow rates for the inlet air and the air delivered to the exposure chamber. We have used the centrifugal concentrator to deliver concentrated ambient urban PM to a nose-only exposure chamber and examined the concentrating effect across ambient particle sizes. PMID- 10380161 TI - Inflammatory response in humans exposed to 2.0 ppm nitrogen dioxide. AB - Nitrogen dioxide (NO2) is a common indoor air pollutant, especially in homes with unvented combustion appliances. Epidemiological studies suggest that children living in homes with unvented heating sources are more prone to respiratory infections than children living in homes with lower levels of NO2. However, experimental studies in which human volunteers were exposed acutely to moderate levels of NO2 (0.5-2.0 ppm) have shown little evidence of lung inflammation or decreased host resistance capacity. In the study reported here, 8 healthy volunteers were exposed to 2.0 ppm NO2 and to filtered air for 4 h while undergoing intermittent moderate exercise. Bronchoalveolar lavage was performed the following morning. The lavage was divided into a predominantly bronchial washing (first 20 ml of lavage; BL) and a predominantly alveolar washing (BAL). In the BL, NO2 exposure caused increases in polymorphonuclear neutrophils (PMNs), interleukin 6 (IL-6), IL-8, alpha1-antitrypsin, and tissue plasminogen activator, and decreases in epithelial cells. In the BAL, there were no NO2-induced changes in either cell numbers or soluble mediators. On the other hand, alveolar macrophages from BAL showed a decrease in the ability to phagocytose unopsonized Candida albicans and a decrease in superoxide production. No difference in susceptibility to virus infection was found between the NO2- and air-exposed macrophages. No changes in lung function were observed, but the aerosol bolus recovery technique revealed a statistically significant (p <.05) decrease in the fraction of aerosol recovered following nitrogen dioxide exposure, which is suggestive of small obstructive changes induced by NO2. PMID- 10380162 TI - Models for mesothelioma incidence following exposure to fibers in terms of timing and duration of exposure and the biopersistence of the fibers. AB - The health effects of inhaled fibers are related to the intensity and duration of exposure and occur many years after the exposure. In particular, the incidence of mesothelioma after exposure to asbestos is proportional to the intensity of exposure (fibers per milliliter of air) and the duration of exposure, and to the time that has elapsed since the exposure. The incidence increases with time since exposure to a power of between 3 and 4. The disease process resulting from exposure to fibers in the air is presumably related to the dose of fibers in the lungs, which depends on the exposure level and duration, and also on the size characteristics of the fibers influencing their inhalation and retention in the lungs. Models incorporating these characteristics have been found to be satisfactory in explaining the incidence of mesothelioma over time after exposure to asbestos. Most of the epidemiological modeling has been for occupational exposure to one of the amphibole asbestos types (crocidolite or amosite), for which heavy exposure produces a high incidence of mesothelioma. Occupational exposure to chrysotile asbestos has resulted in a much lower incidence of mesothelioma. Crocidolite asbestos is much more biopersistent than chrysotile asbestos in the sense that after retention in the lungs it is eliminated only slowly (half-time of several years). If fibers are eliminated then the dose in the lungs declines following exposure, and this may influence the disease process. This concept is more important for synthetic mineral fibers, such as glass wool, which are used as a substitute for asbestos. These fibers are much less biopersistent than asbestos, with half-times of weeks or even days. Biopersistence is related to the dissolution of fibers. This is a physical chemical process that may be expected to proceed at about the same rate in rats and humans. The predicted effect of biopersistence of fibers has been explored using the basic mesothelioma incidence model generalized to include a term representing exponential elimination over time. The influence of solubility of fibers on the mesothelioma rate is 17 times higher in humans than in rats. This is because rats are aging and developing cancer at a much quicker rate than humans, and hence the influence of dissolution is less. Thus, the predicted mesothelioma incidence in humans is highly dependent on the rate of elimination across the range covering asbestos and the more durable synthetic fibers, but in rats a similar dependence occurs at a 17 times higher rate of elimination corresponding to the less durable synthetic fibers. The possible carcinogenic effects of fibers are often determined from animal experiments, but these results suggest that the extrapolation from rats to humans is highly dependent on the biopersistence of fibers, in the situation where the elimination is through dissolution of fibers at a rate independent of species and the speed of the cancer process is species dependent. This implies that relatively soluble fibers that do not produce disease in rat experiments are even less likely to produce disease in humans. PMID- 10380163 TI - Acute biological effects of intratracheally instilled titanium dioxide whiskers compared with nonfibrous titanium dioxide and amosite in rats. AB - The dimensions of man-made mineral fiber whiskers are similar to those of some kinds of asbestos. Thus these mineral fibers raise the concern for potential health hazard for workers exposed in the occupational environments. This study was designed to define acute biological effects of intratracheally administered titanium dioxide whiskers (TO1) compared with nonfibrous titanium dioxide (TOP) and UICC amosite (Ams), and their relations to acute lung inflammation in rats. The observed geometric mean length (microm) and width (microm) and geometric standard deviation are: TO1(2.1[2.0], 0.14[1. 53]); Ams (4.3[3.3], 0.31[1.9]); and TOP (50 nm, 1-2 microm aggregates). Ten-week-old Wistar-Jcl male rats received a single tracheal injection of test materials at doses between 0.05 and 1.0 mg/rat. Control animals were injected with the same volume of saline. Lung tissue and bronchoalveolar lavage (BAL) fluid were collected from rats on days 1, 3, and 7 after administration. In the group injected with TO1, total protein, cytokine-induced neutrophil chemoattractant (CINC)/growth-regulated gene product (GRO), interleukin (IL) 1beta, and tumor necrosis factor (TNF) alpha increased on day 1. Subsequently, total elastolytic activity and fucose levels in BAL increased by day 3. All parameters, except for fucose in BAL, recovered to the normal levels. Animals in the Ams group showed increased total protein and CINC/GRO and decreased total elastolytic activity in a dose-dependent manner on day 1. The fucose level increased on day 3 in the Ams group. All parameters returned to their control levels on day 7. Animals in the TOP group did not show significant changes any of parameters during the experimental period. Gene expression of TNF-alpha and monocyte chemoattractant protein (MCP) 3 in the lung increased dose-dependently in the animals treated with the three materials. The mRNAs for eotaxin and MIP-1alpha were overexpressed in the lung of animals treated with Ams and TO1, while RANTES mRNA was overexpressed dose-dependently in the lung of animals treated with Ams on day 1. Onset of inflammatory response was more rapid in the Ams group than the TO1 group. Recovery of the fucose level in BAL was slower in the TO1 group than in the Ams group, though we observed similar histopathological changes in the lung of animals with TO1 or Ams. We conclude that whisker-induced acute biological effects in the lung may be related to the shape of the whiskers and not to their chemical composition or surface crystal structure, showing biological effects similar to those of UICC amosite. PMID- 10380164 TI - Comparison of particle lung doses from the fine and coarse fractions of urban PM 10 aerosols. AB - The U.S. Environmental Protection Agency (EPA) recently revised the national ambient air quality standards to include a new PM-2.5 particulate standard. We examine the contributions of fine (PM-2.5) and coarse (PM-2.5 to -10) fraction of typical urban aerosols to particle doses in different lung airways resulting from 24-h exposure to the standard concentration of 150 microg m-3. The aerosol is assumed to have a bimodal lognormal mass distribution with mass median diameters of 0.2 and 5 microm, and geometric standard deviation of 1.7 and 57% of the mass in the fine (PM-2.5) mode. The daily mass dose from exposure to 150 microg m-3 of PM-10 in the nasopharyngeal (NPL) region is 20-51 microg day-1 (1.5% of inhaled fines) and 377-687 microg day-1 (30% of inhaled coarse), respectively, of fine and coarse mass filtered in the nose. Similar daily mass doses from fine and coarse fractions, respectively, to the tracheobronchial (TBL) region are 28-38 (1.5%) and 40-52 (4%) microg day-1 and to the pulmonary (PUL) region are 18-194 (6%) and 32-55 microg day-1 (2%). The daily number dose in the NPL region is 5-15 x 10(8) (0.06% of inhaled fines) and 5-10 x 10(6) day-1 (13% of inhaled coarse) respectively, of fine and coarse particles. Similar number doses to the TBL region are 2.2-3.1 x 10(10) (2%) and 7.1-11. 1 x 10(5) (2%) day-1 and to the PUL region are 1.6-16.7 x 10(10) (9%) and 2.9-17.0 x 10(5) (3%) day-1. The daily surface mass dose (microg cm-2 day-1) from coarse fraction particles is large in generations 3-5. The daily number dose (particles day-1) and surface number dose (particles cm-2 day-1) are higher from the fine than the coarse fraction, by about 10(3) to 10(5) times in all lung airways. Fine fraction particles result in 10,000 times greater particle number dose per macrophage than coarse fraction particles. Particle number doses do not follow trends in mass doses, are much larger from fine than coarse fraction, and must be considered in assessing PM health effects. For the assumed fine fraction ratio of 0.57, the estimated increase in protection from the new PM-2.5 standards is a 25% and 47% lower dose, respectively, at the 24-h and annual standard in comparison with the respective PM-10 standards. The mass fraction in the fine mode depends upon the local sources, will vary with different extents of control of various source types, and will influence the choice of control strategy to meet the revised standard. PMID- 10380165 TI - The importance of the diluent for airway transport of toluene diisocyanate following intranasal dosing of mice. AB - Uncertainty of the transport of reactive chemicals to the lung is a major concern when using intranasal dosing of animals. In a preliminary study using mice, intranasal instillation of the dyes methylene blue (in water) and Sudan black B (in 1:4 ethyl acetate:olive oil), indicated that the following conditions were necessary to achieve transport to the lung: (1) aqueous diluent, (2) light anesthesia prior to dosing, (3) holding the animal in a supine position during chemical application, and (4) maintaining the animal in the same position postdosing. Using these conditions, we investigated the distribution of toluene diisocyanate (TDI), a major industrial asthmogen, to the lung following intranasal administration. Female C57BL/6 mice received 20 microl of 1% TDI in ethyl acetate:olive oil (1:4). Group 1 received a single application on day 1; group 2, single applications on 2 consecutive days; group 3, single applications on 4 consecutive days; and group 4, a single application of the vehicle on 2 consecutive days. All mice were necropsied 24 h after the final application. The nasal passages, upper pharynx, trachea, lungs, and olfactory bulbs of each animal were examined with hematoxylin-eosin and immunohistochemical staining, the latter using a rabbit anti-TDI antiserum. Histopathology revealed desquamation of ciliated epithelial cells as well as inflammatory cell debris in the nasal cavity and upper pharynx of animals in groups 1-3. The intensity of these changes was dependent on the number of applications. No inflammation was observed in the trachea, lungs, or olfactory bulbs in any of the groups. Immunohistochemical examination revealed positive staining for the TDI moiety in epithelial cells of the nasal cavity and upper pharynx in animals of groups 1-3. No staining was observed in the trachea, lungs, or olfactory bulbs of any animal. These results suggest that TDI, when dissolved in olive oil:ethyl acetate and applied intranasally, does not reach the trachea and/or lower airways. PMID- 10380166 TI - Respiratory hypersensitivity to diphenylmethane-4,4'-diisocyanate in guinea pigs: comparison with trimellitic anhydride. AB - Published evidence demonstrates successful induction and elicitation of respiratory hypersensitivity in guinea pigs by the known human respiratory allergens trimellitic anhydride (TMA) and diphenylmethane-4,4'-diisocyanate (MDI). From these data it is apparent that TMA-related respiratory hyperresponsiveness can be elicited readily in guinea pigs upon inhalation challenge with the free chemical. Despite the interlaboratory variability in methodological procedures used for the sensitization as well as elicitation of response and the wide range of concentrations of TMA employed for challenge exposures (6-57 mg/m(3) air), TMA had been unequivocally identified as a benchmark respiratory sensitizer by measurements of the respiratory rate during challenge. The protocols were duplicated to examine the respiratory sensitizer MDI. In intradermally sensitized guinea pigs, changes in immediate-onset-like respiratory response were observed when MDI challenge concentrations exceeded approximately 30 mg MDI/m(3) air. Collective experimental evidence suggests that the respiratory responses observed upon challenge with TMA were markedly more pronounced and easier to identify than those recorded following challenge with MDI or MDI conjugate. In contrast to TMA, irritant concentrations of MDI had to be used to elicit any respiratory response and the differentiation of irritant and allergic responsiveness became increasingly difficult. Despite the absence of unequivocal changes in breathing patterns upon MDI challenge, MDI-sensitized animals displayed elevated anti-MDI immunoglobulin G1 (IgG1) antibodies, and a significant influx of eosinophilic granulocytes in the bronchial wall and lung associated lymph nodes. Therefore, it is believed that the robustness of this animal model to identify low-molecular-weight agents as respiratory sensitizer is increased when several endpoints are considered. These are (1) positive respiratory response upon challenge with the hapten, and if negative, also challenge with the conjugate of the hapten; (2) an influx of eosinophilic granulocytes; and (3) increased specific IgG1 response. Furthermore, it appears that particles in the range of approximately 2-6 microm evoke more consistent respiratory response upon challenge exposure than particles in the 1-2 microm range. PMID- 10380168 TI - A dosimetry model of nickel compounds in the rat lung. AB - Experimental data from inhalation studies in rats were used to develop mathematical models of deposition, clearance, and retention kinetics for inhaled Ni compounds (high-temperature [green] NiO, Ni3S2, and NiSO4*6H2O) in the rat lung. For deposition, an updated version of an earlier model (Yu & Xu, 1986) was used in this study. Three major mechanisms of airway deposition-impaction, sedimentation, and diffusion-were considered in the deposition model. In the development of a clearance model, a single compartment model in the lung was used and a general assumption was made that the clearance of the insoluble and moderately soluble nickel compounds (high-temperature [green] NiO and Ni3S2, respectively) depends highly on the volume of retained particles in the lungs. For the highly soluble nickel compound (NiSO4 *6H2O), the clearance rate coefficient was assumed to depend on the retained particle mass and total alveolar surface. The retention half-time, however, was found to increase with the lung burden for high-temperature (green) NiO and NiSO4*6H2O particles but decrease with the lung burden for Ni3S2 particles. PMID- 10380169 TI - Adaptive and non-adaptive responses in rats exposed to ozone, alone and in mixtures, with acidic aerosols. AB - Healthy young adult (300 g) Sprague-Dawley rats were exposed for 1-day or 5-day periods, nose only, to purified air (CA) or four different pollutant atmospheres. Pollutant atmospheres included (a) 0.2 ppm ozone; (b) 0.4 ppm O3; (c) a low concentration mixture of ozone and sulfuric acid-coated carbon particles (0.2 ppm, 100 microg/m(3) and 50 microg/m(3), respectively); and (d) a high concentration O3 and sulfuric acid-coated carbon particle mixture (0.4 ppm, 500 microg/m(3) and 250 microg/m(3), respectively). Following 1-day exposures to the high O3 concentration, significant (p< or =.05) decreases were observed in respiratory tidal volumes and significant increases were observed in lung inflammatory response. Following 5-day exposures to 0.4 ppm ozone, tidal volumes and lung inflammation were not significantly different from those seen in CA controls. In contrast, following 5-day exposures to the high-concentration O3 particle mixture, lung inflammation was increased significantly relative to that seen after 1-day high concentration mixture exposure or after CA exposure. Macrophage Fc-receptor binding, an important immunological function of macrophages, was significantly depressed after 5-day exposures to either the high or low-concentration O3-particle mixtures compared to 1-day exposures or to CA. Thus, at the concentrations tested, repeated exposures to O3 produced diminished responses in breathing pattern changes and lung parenchymal injuries compared to acute, single exposures. This diminution was not observed after exposures to mixtures of acidic particles plus ozone. We conclude that mixtures of ozone and acidic particles may alter adaptive mechanisms that have been reported by us and others after repeated exposures to ozone alone. PMID- 10380167 TI - Comparative mutagenic dose of ambient diesel engine exhaust. AB - Diesel engine exhaust contains carbon-based particles that can be inhaled and deposited on lung surfaces. Concern about the carcinogenic potential of diesel engine exhaust derives in part from the mutagenic activity of organics that can be extracted from exhaust particles. However, the lung cancer risk is controversial, and diesel exhaust is a candidate for further evaluation. A comparative potency approach can be used to rank the mutagenic risk of diesel exhaust with other combustion products. We compared the specific mutagenic activities of cigarette smoke condensate (CSC) and diesel exhaust particle extract (DEPE) and estimated "mutagenic dose" to the lungs. Although the specific mutagenic activities of CSC and DEPE are similar in magnitude, it is the dose reaching the lungs that is more relevant for comparing mutagenic potential. We calculated that, depending on the source of CSC and DEPE, a person would have to inhale approximately 63 to 181 mg of particulate from diesel engine exhaust to match the mutagenic dose of 1 cigarette. We also calculated that a person would have to breathe diesel exhaust (1.5 microg/m(3), estimated total personal exposure) for 6 to 16 yr to equal the mutagenic dose of 1 cigarette. Although instructive, comparative potency results should be used cautiously due to the need for simplifying assumptions. For example, the type of mutagenic assay and the source of cigarettes and diesel engine exhaust could affect dose estimates to some degree; however, the extent to which diesel particle mutagens are bioavailable would have an even greater effect on estimates of relative risk. For both cigarette smoke and diesel exhaust particles, we assumed that the organic mutagens are 100% bioavailable. In summary, our analysis showed a larger mutagenic dose-to-target-tissues in the smoke of one cigarette as compared to a year of exposure to diesel exhaust particulate at ambient levels. PMID- 10380170 TI - POCK model simulations of pulmonary quartz dust retention data in extended inhalation exposures of rats. AB - In recent years, a physiology-oriented multicompartmental kinetics (POCK) model was developed to simulate pulmonary retention data of biopersistent, noncytotoxic aerosols in long-term inhalation exposures of rats. Experimental data were successfully simulated for submicrometer-sized aerosols like carbon black, diesel soot, and titanium dioxide and for a micrometer-sized xerographic toner aerosol (Stober et al., 1994, 1995). This article describes for various rat strains successful POCK model simulations of experimental pulmonary retention data of micrometer-sized aerosols of biopersistent cytotoxic SiO2 modifications like quartz and quartzite. In the past, the POCK model was not applied to cytotoxic aerosols and dusts. Cytotoxicity was considered incompatible with the model assumption of a constant macrophage lifetime independent of the macrophage aerosol load. The few relevant experimental retention studies with biopersistent silica found in the open literature showed particulate lung burdens up to some 15 mg per rat lung. Apparently, at these loads, pulmonary burdens could be simulated because the fraction of alveolar macrophages killed by the cytotoxic particles was possibly still small compared to the total number of viable macrophages. Of necessity, however, the classical alveolar clearance in these studies was exclusively performed by alveolar macrophages that were burdened with cytotoxic particles, and the cells appeared to suffer from a substantial initial decrease of their inherent mobility. Thus a sizeable reduction of the alveolar clearance rate coefficient in comparison to nontoxic aerosol was found. The results for the model parameters of several different exposure studies are shown and interpreted in comparison to nontoxic titanium dioxide retention parameters. PMID- 10380171 TI - Sulfate content correlates with iron concentrations in ambient air pollution particles. AB - Current levels of air pollution particles in American cities can increase human mortality. Both the mechanism of injury and the responsible components are not known. We have postulated that injury following air pollution particle exposure is produced through a generation of oxygen-based free radicals catalyzed by metals present in the particles. As a result of its abundance in the atmosphere, sulfate appears to potentially be the most successful ligand to complex metal cations. We tested the hypothesis that (1) some portion of iron in ambient air pollution particles is present as sulfate and (2) this relationship between iron and sulfate results from the capacity of the latter to function as a ligand to mobilize the metal from the oxide. Concentrations of sulfate and iron in acid extracts of 20 filters (total suspended particles) from Utah were measured using inductively coupled plasma emission spectroscopy. In vitro oxidant generation was also measured using thiobarbituric acid-reactive products of deoxyribose. There were significant correlations between sulfate content, iron concentrations, and oxidant generation. Agitation of calcium sulfate with iron(III) oxide produced concentrations of water-soluble, catalytically active iron. We conclude that some portion of iron in the atmosphere is present as a sulfate. This relationship between sulfate and iron concentrations is likely the product of SO42- functioning as a ligand for the meal after its mobilization from an oxide by photoreduction. There were also associations between sulfate content, iron concentrations, and oxidant generation. However, sulfates had no capacity to support electron transport unless they were present with iron. PMID- 10380172 TI - Interspecies differences in time course of pulmonary toxicity following repeated exposure to ozone. AB - To compare the extent and time course of pulmonary injury and repair in 3 rodent species, rats, mice and guinea pigs were continuously exposed for 3, 7, 28, and 56 days to 400 and 800 microg O3/m(3) (0.2 and 0.4 ppm). Recovery from 28 days of exposure was studied at 3, 7, and 28 days after exposure. Pulmonary injury and repair was studied at various time points by histology, electron microscopy, morphometry, and biochemistry. In all 3 species a concentration-related centriacinar inflammation occurred, with a maximum after 3 days of exposure. The number of alveolar macrophages and the pulmonary cell density in the centriacinar region increased progressively until 56 days of exposure, with the guinea pig the most sensitive species. Only the mouse displayed a concentration and exposure time dependent hypertrophy of bronchiolar epithelium. After 56 days of exposure to 800 microg O3/m(3) in the rat and the guinea pig, giant lamellar bodies in type II cells were present. Exposures for 3 and 7 days at near ambient ozone concentrations (400 microg O3/m(3)) resulted in significantly elevated lung enzyme activities in the mouse, and in significant histological and morphometric changes in all 3 species. In rat and guinea pigs exposures for 56 days resulted in alveolar duct fibrosis. The highest biochemical response and the slowest recovery from ozone exposure were seen in the mouse. Histology, morphometry, and biochemistry revealed a total recovery from a 28-day exposure period in rats after 28 days, while in guinea pigs the ductular septa were still thickened and in mice all enzyme activities were still elevated in comparison with control values. In conclusion, the response of mice to ozone was evaluated as most severe, followed by those of guinea pigs and least in rats. PMID- 10380173 TI - Detection of ozone-induced DNA single strand breaks in murine bronchoalveolar lavage cells acutely exposed in vivo. AB - Single-strand breaks (SSBs) in DNA have been used a biomarker of oxidative damage. The comet assay, also known as single-cell gel electrophoresis, was used to investigate the ability of ozone (O(3)) to induce DNA SSBs in murine bronchoalveolar lavage (BAL) cells. The comet assay is more sensitive than other techniques currently utilized for detecting SSBs and requires fewer cells. In the present study, 3 mice were exposed for 3 h to 0.25 ppm of O(3), and 3 to 0.5 ppm of O(3) for 3 h. Two air-exposed mice served as negative controls. All mice were euthanized 3 h after exposure, at which time BAL cells were recovered from the lungs and stained with ethidium bromide. BAL cells recovered from an air-exposed mouse were exposed to various concentrations of H(2)O(2) in vitro for 1 h at 4 degrees C. Excluding cells from the H(2)O(2) group (n = 25), 50 randomly selected BAL cells were graded by comet tail length into 1 of 4 categories: no damage (0 mm), low damage (1-10 mm), medium damage (11-30 mm), and high damage (31 + mm). The nonparametric Wilcoxon rank-sum test was used for statistical analysis, and p values lower than .05 were considered significant. The H(2)O(2) and the 0.25 and 0.5 ppm O3 groups showed statistically significant increases in DNA SSBs as compared to air-exposed controls. The results of this study indicate that (1) O(3) induces DNA strand breaks in murine BAL cells at 0.25 and 0.5 ppm, as evidenced by statistically significant increases in the length of comet tails for O(3)-exposed groups, and (2) the comet assay can be used to assess O(3)-induced SSBs for in vivo exposures. Therefore, it has the potential as a biomarker for in vivo oxidant exposures. PMID- 10380174 TI - Health effects of sulfur-related environmental air pollution. I. Executive summary. AB - The motivation of simulating real-world environmental exposure in a number of long-term studies with dogs was to address the question of whether or not perpetual inhalation of air pollutants can initiate diseases in healthy lungs and can thus contribute to the increasing prevalence of respiratory diseases in industrialized countries. The major conclusion of this article is that this question has to be answered in the negative for the simultaneous inhalation of the major constituents of combustion-related air pollution, particle-associated sulfur(IV), and particle-associated hydrogen ions. Over 13 mo, 8 healthy beagle dogs were exposed in 2 whole-body chambers daily for 16.5 h to 1 microm neutral sulfite [sulfur(IV)] particles at a mass concentration of 1.5 mg m-3 and for 6 h to 1.1 microm acidic sulfate particles carrying 15 micromol m-3 hydrogen ions into the canine lungs. This longitudinal study was characterized by repeated observations of individual respiratory response patterns. To establish baseline data the dogs were repeatedly examined preexposure while the chambers were ventilated over 16 mo with clean air. Each individual served thus as its own control. Another eight dogs served as additional controls. They were housed in 2 chambers ventilated with clean air over the entire study period of 29 mo. To assess response patterns, respiratory lung function tests were performed pre- and postexposure, segmental lung lavages were repeatedly performed to obtain epithelial lining fluid from the lungs for analysis of cell content, cell function, and biochemical indicators of lung injury, and radiolabeled test particles were used to study pathways of intrapulmonary particle elimination. At the end of the study, the lungs of all animals were morphologically and morphometrically examined. Functional and structural responses were finally compared to those observed previously as a result of a sole exposure of canine lungs to neutral sulfite particles over 10 mo (Heyder et al., 1992). Interactions between responses induced by neutral sulfite and acidic sulfate particles occurred, but antagonism rather than synergism was observed. The responses induced by sulfur(IV) were less pronounced, not detectable, or even reversed when hydrogen ions were also delivered to the lungs. On the other hand, responses not induced by the sole exposure to sulfur(IV) were observed: The activity of alkaline phosphatase was elevated and type II pneumocytes proliferated. It can, however, be concluded that long-term exposure of healthy lungs to particle associated neutral sulfur(IV) and hydrogen ions at concentration close to ambient levels causes subtle respiratory responses but does not initiate pathological processes in the lungs. In other words, the perpetual inhalation of sulfur(IV) and hydrogen ions from the atmospheric environment presents no health risk to the healthy lungs. It is thus also very unlikely that respiratory diseases can be initiated by the inhalation of these pollutants. PMID- 10380175 TI - Health effects of sulfur-related environmental air pollution. II. Cellular and molecular parameters of injury. AB - Recently, concern has been raised about effects related to environmental sulfur and/or acidic aerosols. To assess long-term effects on nonrespiratory lung function, 8 beagle dogs were exposed over a period of 13 mo for 16.5 h/day to a neutral sulfite aerosol at a sulfur(IV) concentration of 0.32 mg m(-3) and for 6 h/day to an acidic sulfate aerosol providing a hydrogen concentration of 15.2 micromol m(-3) for inhalation. Prior to exposure the dogs were kept under clean air conditions for 16 mo to establish physiological baseline values for each animal. A second group of eight dogs (control) was kept for the entire study under clean air conditions. No clinical symptoms were identified that could be related to the combined exposure. Biochemical and cellular parameters were analyzed in sequential bronchoalveolar lavage (BAL) fluids. The permeability of the alveolo-capillary membrane and diethylenetriaminepentaacetic acid (DTPA) clearance was not affected. Similarly, oxidant burden of the epithelial lining fluid evaluated by levels of oxidation products in the BAL fluid protein fraction remained unchanged. Both the lysosomal enzyme beta-N-acetylglucosaminidase and the alpha-1-AT were increased (p <.05). In contrast, the cytoplasmic marker lactate dehydrogenase remained unchanged, indicating the absence of severe damages to epithelial cells or phagocytes. Various surfactant functions were not altered during exposure. Three animals showed elevated levels of the type II cell associated alkaline phosphatase (AP), indicating a nonuniform response of type II cells. Significant correlations were found between AP and total BAL protein, but not between AP and lactate dehydrogenase, suggesting proliferation of alveolar type II cells. Absolute and relative cell counts in the BAL fluid were not influenced by exposure. Alveolar macrophages showed no alterations with regard to their respiratory burst upon stimulation with opsonized zymosan. The percentage of alveolar macrophages capable of phagocytozing latex particles was significantly decreased (p<.05), while the phagocytosis index was not altered. In view of the results of this and previous studies, we conclude that there is no synergism of effects of these two air pollutants on nonrespiratory lung functions. It is hypothesized that antagonistic effects of these air pollutants on phospholipase A2-dependent pathways account for compensatory physiological mechanisms. The results emphasize the complexity of health effects on lung functions in response to the complex mixture of air pollutants and disclose the precariousness in the risk assessment of air pollutants for humans. PMID- 10380177 TI - Health effects of sulfur-related environmental air pollution. IV. Respiratory lung function. AB - Recently concern has been raised about health effects related to environmental sulfur and/or acidic aerosols. To assess long-term effects on respiratory lung function, 8 beagle dogs were exposed over a period of 13 mo for 16.5 h/day to 1 microm neutral sulfite aerosol with a particle-associated sulfur(IV) concentration of 0.32 mg m(-3) and for 6 h/day to 1.1-microm acidic sulfate aerosol providing an hydrogen ion concentration of 15.2 micromol m(-3) for inhalation. Prior to exposure the dogs were kept under clean air conditions for 16 mo to establish physiological baseline values for each dog. A second group of eight dogs (control) was kept for the entire study under clean air conditions. Before and at the end of exposure, respiratory lung function was evaluated in both groups in anesthetized and mechanically ventilated animals. Lung volumes as well as static and dynamic lung compliances were measured. Series dead-space volumes and slopes of the alveolar plateau for respiratory (O2, CO2) and inert test gases (He, SF6) were determined from single-breath washout tracings. Monodisperse 0.9-microm DEHS droplets were used to assess convective mixing in the lungs and to evaluate airway dimensions in vivo. Gas exchange across the alveolar-capillary layer was characterized by membrane diffusing capacity for carbon monoxide and alveolar-arterial pressure differences for respiratory gases. A bronchial challenge with carbachol was used to assess airway responsiveness. In comparison to the control group, dogs exposed to sulfur(IV) and acidic aerosol exhibited no significant changes in any respiratory lung function parameter. Also the responsiveness of the bronchial airways to carbachol was not affected. In view of the results obtained in this and previous studies, we conclude that anticipated synergistic effects of the two air pollutants on pulmonary lung function were not observed. It is hypothesized that antagonistic effects of the air pollutants on the activity of phospholipase A2 play an important role and account for counteracting physiological compensatory mechanisms. The results emphasize the complexity of health effects on lung function in response to the complex mixtures of ambient air pollutants and witness the precariousness in the risk assessment of air pollutants for humans. PMID- 10380176 TI - Health effects of sulfur-related environmental air pollution. III. Nonspecific respiratory defense capacities. AB - Recently concern has been raised about health effects related to environmental sulfur and/or acidic aerosols. To assess long-term effects on respiratory lung function, 8 beagle dogs were exposed over a period of 13 mo for 16.5 h/day to 1.0 microm neutral sulfite aerosol with a particle associated sulfur(IV) concentration of 0.32 mg m(-3) and for 6 h/day to 1.1 microm acidic sulfate aerosol providing an hydrogen ion concentration of 15.2 micromol m(-3) for inhalation. Prior to exposure the dogs were kept under clean air conditions for 16 mo to establish physiological baseline values for each dog. A second group of eight dogs (control) was kept for the entire study under clean air conditions. Nonspecific defense mechanisms in the airways and in the peripheral lung were studied during chronic exposure of the combination of neutral sulfur(IV) and acidic sulfur(VI) aerosols. No functional changes of tracheal mucus velocity were found, in agreement with unchanged morphometry of the airways. However, the exposure resulted in changes of several alveolar macrophage (AM) mediated particle clearance mechanisms: (1) Based on in vivo clearance analysis and cultured AM studies using moderately soluble cobalt oxide particles, intracellular particle dissolution was significantly reduced since phagolysosomal proton concentration was decreased. We deduce exposure-related malfunction of proton pumps bound to the phagolysosomal membrane as a result of an increase of cytosolic proton concentration. (2) Based on in vivo clearance analysis using insoluble polystyrene particles, AM-mediated particle transport from the lung periphery toward ciliated terminal bronchioli and further to the larynx was significantly reduced. Activation of epithelial type II cells at the entrance of alveoli was inferred from observed type II cell proliferation at those alveolar ridges and enhanced secretion of alkaline phosphatase in the fluid of bronchoalveolar lavages. As a result, hypersecretion of chemotactic mediators by activated type II cells at these loci led to the observed decrease of particle transport toward ciliated bronchioli. (3) Based on in vivo clearance analysis using insoluble polystyrene particles, particle transport from the alveolar epithelium into interstitial tissues was increased and (4) particle transport to the tracheobronchial lymph nodes was significantly enhanced. Particle transport into interstitial tissues is the most prominent clearance pathway from the canine alveolar epithelium. We conclude that the deteriorated particle transport toward ciliated terminal bronchioli resulted in an enhanced particle transport across the epithelial membrane into interstitial tissues and the lymphatic drainage. The observed alterations in alveolar macrophage-mediated clearance mechanisms during chronic exposure of these air pollutants indicate an increased risk of health. PMID- 10380179 TI - Human health risk assessment: a historical overview and alternative paths forward. AB - Risk assessment has become a more structured activity during the past 50 years and increasingly is being used to inform major policy decisions. Much use has been made of the hazard identification phase of risk assessment to identify potentially hazardous materials or situations and guide actions to minimize potential risks. Much less frequently the process has been carried further, with estimates developed of the potency of the hazardous agent for causing adverse effects. And even less frequently, robust estimates of exposure have been developed. Thus, in only a few instances have risks been fully characterized in quantitative terms for either individuals or populations. To develop scientifically valid risk characterizations for many chemicals, much more scientific information must be acquired in a targeted manner to establish the potency of chemicals for causing cancer or other adverse health effects. Similar substantial effort must also be applied to characterizing the exposure populations receive from specific chemicals released from various source categories. In the absence of these scientifically rigorous approaches it is likely that societal actions will be guided primarily by identification of potential hazardous agents, with attempts made to minimize the hazard by banning or restricting use of the agent. This precautionary approach may not yield the maximum reduction in health risks to society for the investments made and, in addition, may deny society access to materials or processes that under appropriate conditions of use would not result in significant health risks and may indeed, have substantial net benefits to society. PMID- 10380178 TI - Health effects of sulfur-related environmental air pollution. V. Lung structure. AB - The lungs of 8 male beagle dogs were examined morphologically and morphometrically after exposure for 13 mo to a respirable sulfur(IV) aerosol at a mass concentration of 1.53 mg m(-3) (16.5 h/day), and to an acidic sulfate aerosol carrying 15.2 micromol m(-3) hydrogen ions into the lungs (6 h/day). An additional eight dogs served as unexposed controls. Standard morphometric analyses of both the surface epithelia of the conducting airways and the alveolar region were performed. These analyses showed no difference between the exposure group and control group. However, there was a tendency to an increase in the volume density of bronchial glands in the exposure group. Five of eight exposed animals showed thickened ridges (knob-like structures) at the entrance to alveoli in the alveolar duct and alveolar sac. Transmission electron microscopy revealed that the thickening was mainly due to type II cell proliferation. As the previous experiment using sulfite aerosol only showed no alterations in the proximal alveolar regions, the changes observed may be considered as effects of acidic sulfate aerosol alone or in combination with sulfite. These findings suggest that sulfur aerosols have the potential to induce epithelial alterations in the proximal alveolar region, which is a primary target for air pollutants. PMID- 10380180 TI - [Validation of the Spanish version of the social adaptation scale in depressive patients]. AB - INTRODUCTION: The Social Adaptation Self-evaluation Scale (SASS) allows a very easy evaluation of the individual's perspective about himself and his environment, together with his behaviour and social motivation. Taking into account that social maladjustment is high prevalent and well documented for depressed subjects and it is one of the most limiting aspects of their global performance, the aim of the present work is to validate the Spanish version of SASS for its application in subjects diagnosed as suffering major depression (DSM IV). METHODOLOGY: A cross sectional multicentre study was carried out by collecting information on 464 patients (34.7% men and 65.3% women range 18 to 65 years), which included their answers to the scale and relevant sociodemographic and clinical variables. RESULTS: The principal component analysis corroborated the identification of 4 main factors: the first one being extrafamily relationships (31.4% of inertia); the second, work and leisure (7.6% of inertia); the third, social and cultural interests (5.6% of inertia); and the fourth factor, family relationships and behavioral strategies (5.5% of intertia). The scale has provided adequate reliability and validity indexes as well as sensitivity to the severity of the depressive episode. CONCLUSIONS: The Spanish version of the SASS has proven to be an adequate instrument for evaluating social adjustment in depressive subjects. However, future studies must corroborate its sensitivity to the effects of antidepressant treatment and potential differences between the antidepressant agents employed. PMID- 10380181 TI - Genetic network analysis in light of massively parallel biological data acquisition. AB - Complementary DNA microarray and high density oligonucleotide arrays opened the opportunity for massively parallel biological data acquisition. Application of these technologies will shift the emphasis in biological research from primary data generation to complex quantitative data analysis. Reverse engineering of time-dependent gene-expression matrices is amongst the first complex tools to be developed. The success of reverse engineering will depend on the quantitative features of the genetic networks and the quality of information we can obtain from biological systems. This paper reviews how the (1) stochastic nature, (2) the effective size, and (3) the compartmentalization of genetic networks as well as (4) the information content of gene expression matrices will influence our ability to perform successful reverse engineering. PMID- 10380182 TI - Identification of genetic networks from a small number of gene expression patterns under the Boolean network model. AB - Liang, Fuhrman and Somogyi (PSB98, 18-29, 1998) have described an algorithm for inferring genetic network architectures from state transition tables which correspond to time series of gene expression patterns, using the Boolean network model. Their results of computational experiments suggested that a small number of state transition (INPUT/OUTPUT) pairs are sufficient in order to infer the original Boolean network correctly. This paper gives a mathematical proof for their observation. Precisely, this paper devises a much simpler algorithm for the same problem and proves that, if the indegree of each node (i.e., the number of input nodes to each node) is bounded by a constant, only O(log n) state transition pairs (from 2n pairs) are necessary and sufficient to identify the original Boolean network of n nodes correctly with high probability. We made computational experiments in order to expose the constant factor involved in O(log n) notation. The computational results show that the Boolean network of size 100,000 can be identified by our algorithm from about 100 INPUT/OUTPUT pairs if the maximum indegree is bounded by 2. It is also a merit of our algorithm that the algorithm is conceptually so simple that it is extensible for more realistic network models. PMID- 10380183 TI - Modeling gene expression with differential equations. AB - We propose a differential equation model for gene expression and provide two methods to construct the model from a set of temporal data. We model both transcription and translation by kinetic equations with feedback loops from translation products to transcription. Degradation of proteins and mRNAs is also incorporated. We study two methods to construct the model from experimental data: Minimum Weight Solutions to Linear Equations (MWSLE), which determines the regulation by solving under-determined linear equations, and Fourier Transform for Stable Systems (FTSS), which refines the model with cell cycle constraints. The results suggest that a minor set of temporal data may be sufficient to construct the model at the genome level. We also give a comprehensive discussion of other extended models: the RNA Model, the Protein Model, and the Time Delay Model. PMID- 10380184 TI - Linear modeling of mRNA expression levels during CNS development and injury. AB - Large-scale gene expression data sets are revolutionizing the field of functional genomics. However, few data analysis techniques fully exploit this entirely new class of data. We present a linear modeling approach that allows one to infer interactions between all the genes included in the data set. The resulting model can be used to generate interesting hypotheses to direct further experiments. PMID- 10380185 TI - Sensitivity of biological models to errors in parameter estimates. AB - Since A. M. Turing's paper proposing a mathematical basis for pattern formation in developing organisms many mathematical approaches have been proposed to model biological phenomenon. Continued laboratory study and recent improvements in measurement capabilities have provided an immense quantity of raw gene expression data. The level of data now available demands the development of well characterized and tested computational tools. Thus, we have examined one mathematical model's sensitivity to errors in estimating its' parameters. Errors in parameter estimation can arise from noise in the laboratory measurements and recasting of laboratory data. We elected to examine the rule-based mathematical model of Mjolsness et al for its' sensitivity to errors in estimated parameters. We have used the technique of sensitivity equations as generally applied in nonlinear systems analysis. PMID- 10380186 TI - Analysis of the stabilizing effect of Rom on the genetic network controlling ColE1 plasmid replication. AB - A stochastic model of ColE1 plasmid replication is presented. It is implemented by using UltraSAN, a simulation tool based on an extension of stochastic Petri nets (SPNs). It allows an exploration of the variation in plasmid number per bacterium, which is not possible using a deterministic model. In particular, the rate at which plasmid-free bacteria arise during bacterial division is explored in some detail since spontaneous plasmid loss is a widely observed empirical phenomenon. The rate of spontaneous plasmid loss provides an evolutionary explanation for the maintainance of Rom protein. The presence of Rom acts to reduce variance in plasmid copy number, thereby reducing the rate of plasmid loss at bacterial division. The ability of stochastic models to link biochemical function with evolutionary considerations is discussed. PMID- 10380187 TI - Simulation of genetic interaction for Drosophila leg formation. AB - The formation of Drosophila wings and legs are major research topics in Drosophila development, and several hypotheses, such as the polar-coordinate model and the boundary model, has been proposed to explain mechanisms behind these phenomena. A series of recent studies have revealed complex interaction among genes involved in establishing three principal axes (A-P, D-V, and P-D) of leg formation. In this paper, we present a simulation system for leg formation, simulating the genes interactions involved. We use this simulator to investigate a mathematical framework of leg formation which is otherwise well-founded from a molecular perspective. Particularly, we focus on the formation of the expression patterns of dpp, wg, dll, dac, al, en, hh and ci genes, which are involved in the development of the third instar Drosophila leg disc. The most interesting part of this research is showing how the coaxial gene expression patterns behind the P-D axis can be formed, and how positional information, as postulated in the polar coordinate model, can be conveyed to each cell. Our results suggest that P-D axis can be formed by a set of genes with different activation thresholds; the process involves different chemical gradients of dpp and wg products, forming a bi-polar contour. Interestingly, this combination of chemical gradients can specify unique positions of cells for the hemisphere, leaving the A-P axis determiner to decide only whether the cells are anterior or posterior. All in all, our so-called Bi Polar Model describes axial formation of the leg disc well. PMID- 10380188 TI - Evolutionary constraint networks in ligand-binding domains: an information theoretic approach. AB - Ligand-binding sites in homologous protein domains can diverge greatly during evolution. This poses a particularly interesting problem in those cases where the ligand-binding site is situated in, or close to, the domain core, or where ligand docking induces dramatic conformational changes. These features are present in many receptors and enzymes; the hormone-binding domain of the nuclear receptors for steroids and retinoids, for example, exhibits both characteristics. It is therefore of great interest to determine how binding sites for diverse ligands evolve in core regions of structurally dynamic domains. Are evolutionary changes locally restricted to the ligand-binding site, or are they distributed throughout the domain? We describe here an information-theoretic approach for the study of covariation between ligand-contacting residues and compensatory mutations that preserve the structural integrity and the conformational dynamics of ligand binding domains. We apply this method to the analysis of the nuclear receptor ligand-binding domain and show that the ligand-contacting residues in the hormone binding pocket are evolutionarily linked to an extensive network of covarying positions. PMID- 10380189 TI - NetWork: an interactive interface to the tools for analysis of genetic network structure and dynamics. AB - We designed a Java applet called NetWork which enables a user to interactively construct and visualize a genetic network of interest, and to and to evaluate and explore its dynamics in the framework of a Boolean network model. NetWork displays the mechanism of gene interactions at the level of gene expression and enables the visualization of large genetic networks. NetWork can serve as an interactive interface to tools for the analysis of genetic network structure and behavior. PMID- 10380190 TI - Modeling regulatory networks with weight matrices. AB - Systematic gene expression analyses provide comprehensive information about the transcriptional response to different environmental and developmental conditions. With enough gene expression data points, computational biologists may eventually generate predictive computer models of transcription regulation. Such models will require computational methodologies consistent with the behavior of known biological systems that remain tractable. We represent regulatory relationships between genes as linear coefficients or weights, with the "net" regulation influence on a gene's expression being the mathematical summation of the independent regulatory inputs. Test regulatory networks generated with this approach display stable and cyclically stable gene expression levels, consistent with known biological systems. We include variables to model the effect of environmental conditions on transcription regulation and observed various alterations in gene expression patterns in response to environmental input. Finally, we use a derivation of this model system to predict the regulatory network from simulated input/output data sets and find that it accurately predicts all components of the model, even with noisy expression data. PMID- 10380191 TI - Prediction and visualization of structural switches in RNA. AB - There are various cases where the biological function of an RNA molecule involves a reversible change of conformation. paRNAss is a software approach to the prediction of such structural switching in RNA. It is based on three hypotheses about the secondary structure space of a switching RNA molecule, which can be evaluated by RNA folding and structure comparison. In the positive case, the predicted structures must be verified experimentally. Additionally, we give an animated visualization of an energetically favourable transition between the predicted structures. paRNAss is available via the Bielefeld Bioinformatics Server. This paper explains the underlying model and shows that the approach performs well in a variety of applications. PMID- 10380192 TI - A comparative analysis of computational motif-detection methods. AB - The detection of motifs within and among families of protein sequences can provide useful information regarding the function, structure and evolution of a protein. With the increasing number of computer programs available for motif detection, a comparative evaluation of the programs from a biological perspective is warranted. This study uses a set of 20 reverse transcriptase (RT) protein sequences to test and compare the ability of 7 different computational methods to locate the ordered-series-of-motifs that are well characterized in the RT sequences. The results provide insight to biologists as to the usage, value, and reliability of the numerous methods available. PMID- 10380193 TI - A probabilistic approach to consensus multiple alignment. AB - We consider the problem of obtaining the maximum a posteriori probability (MAP) estimate of a consensus ancestral sequence for a set of DNA sequences. Our maximization method, called ASA (dnA Sequence Alignment), can be applied to the refinement of noisy regions of a DNA assembly, to the alignment of genomic functional sites, or to the alignment of any set of DNA sequences related by a star-like phylogeny. Along with the optimal consensus, ASA finds suboptimal solutions together with their relative probabilities. The probabilistic approach makes it possible to establish the limits to which an ancestor can in principle be recovered from diverged sequences. In simulations on rather short synthetic sequences (of length up to 80) with different coverage and error rates ranging from 5% to 30%, ASA restored the consensus from noisy observations essentially as best as is theoretically possible for the given error rates. We also illustrate the performance of ASA on the alignment of E.Coli promoters and the Alu-Sb subfamily of human repeat sequences. Since our model is a special case of a profile HMM, we give a comparison between these two approaches, as well as with other DNA alignment methods. PMID- 10380194 TI - The effects of ordered-series-of-motifs anchoring and sub-class modeling on the generation of HMMs representing highly divergent protein sequences. AB - Hidden Markov Models (HMMs) provide a flexible method for representing protein sequence data. Highly divergent data require a more complex approach to HMM generation than previously demonstrated. We describe a strategy of motif anchoring and sub-class modeling that aids in the construction of more informative HMMs as determined by a new algorithm called a stability measure. PMID- 10380195 TI - Clustering and detection of 5' splice sites of mRNA by k weight-matrices model. AB - We propose a novel method to detect 5' splice sites of eukaryotic mRNA. We have grouped the 5' splice splice sites into various classes. The clustered sites are represented by a set of PWMs. The clustering algorithm is similar to k-means clustering algorithm but the distance definition and the training score function were arranged. The clustered PWMs were applied to 5' splice site detection. The results showed an improvement in comparison with traditional single PWM. The result of the clusters suggests there are new motifs of 5' splice sites. PMID- 10380196 TI - Use of BONSAI decision trees for the identification of potential MHC class I peptide epitope motifs. AB - Recognition of short peptides of 8 to 10 mer bound to MHC class I molecules by cytotoxic T lymphocytes forms the basis of cellular immunity. While the sequence motifs necessary for binding of intracellular peptides to MHC have been well studied, little is known about sequence motifs that may cause preferential affinity to the T cell receptor and/or preferential recognition and response by T cells. Here we demonstrate that computational learning systems can be useful to elucidate sequence motifs that affect T cell activation. Knowledge of T cell activation motifs could be useful for targeted vaccine design or immunotherapy. With the BONSAI computational learning algorithm, using a database of previously reported MHC bound peptides that had positive or negative T cell responses, we were able to identify sequence motif rules that explain 70% of positive T cell responses and 84% of negative T cell responses. PMID- 10380197 TI - Applications of knowledge discovery to molecular biology: identifying structural regularities in proteins. AB - In recent years, there has been an explosive amount of molecular biology information obtained and deposited in various databases. Identifying and interpreting interesting patterns from this massive amount of information has become an essential component in directing further molecular biology research. The goal of this research is to discover structural regularities in protein sequences by applying the Subdue discovery system to databases found in the Brookhaven Protein Data Bank. In this paper we report the results of applying Subdue to several classes of protein structures and discuss the potential significance of these results to the study of proteins. PMID- 10380198 TI - Time and memory efficient algorithm for extracting palindromic and repetitive subsequences in nucleic acid sequences. AB - Genomic science and structural biology meet in the relationship between the sequence and the structure of nucleic acids. The structure that supports each function is preserved in the process of evolution as specific sequences. Particularly, the same sequence which appears in a different place such as a palindromic or repetitive sequence has biophysical meaning: recognition site of dimers, forming stem-loops, and contributions to global structure of nucleic acids. Also, the genetic network, transduction pathway, and tissue specificity largely depend on these. Although the relationship between them can be found experimentally, there is increasing demand for automated analysis. Especially, it is desirable to extract the same character sequences of arbitrary length (especially, very long ones) which co-occur at an arbitrary separation. We propose an algorithm to identify the maximum match sequence at each position with a calculation cost of O(N log N) and memory space of O(N). Applying it to some sequences, we found unexpectedly large palindromes and repeats in DNA. PMID- 10380199 TI - Effective query filtering for fast homology searching. AB - To improve the accuracy of rapid homology searching it is common practice to filter all queries to mask low complexity regions prior to searching. We show in this paper, through a large-scale study of querying the PIR database, that applying popular filtering techniques unselectively to all queries may reduce retrieval effectiveness. We also show that masking queries with our new technique, cafefilter, which uses the overall distribution of motifs in a database, is at least as effective as current popular query filtering tools in large-scale tests. PMID- 10380200 TI - The virtual cell. AB - This paper describes a computational framework for cell biological modeling and simulation that is based on the mapping of experimental biochemical and electrophysiological data onto experimental images. The framework is designed to enable the construction of complex general models that encompass the general class of problems coupling reaction and diffusion. PMID- 10380201 TI - Extension of refractoriness in a model of cardiac defibrillation. AB - This simulation study presents an inquiry into the mechanisms by which a strong electric shock halts life-threatening cardiac arrhythmias. It examines the "extension of refractoriness" hypothesis for defibrillation which postulates that the shock induces an extension of the refractory period of cardiac cells thus blocking propagating waves of arrhythmia and fibrillation. The present study uses a model of the defibrillation process that represents a sheet of myocardium as a biodomain with unequal anisotropy ratios. The tissue consists of curved fibers in which spiral wave reentry is initiated. The defibrillation shock is delivered via two line electrodes that occupy opposite tissue boundaries. Simulation results demonstrate that a large-scale region of depolarization is induced throughout most of the tissue. This depolarization extends the refractoriness of the cells in the region. In addition, new wavefronts are generated from the regions of induced hyperpolarization that further restrict the spiral wave pathway and cause its termination. PMID- 10380202 TI - Location of repetitive regions in sequences by optimizing a compression method. AB - Suppose that a biologist wishes to study some local property P of genetic sequences. If he can design (with a computer scientist) an algorithm C which efficiently compresses parts of the sequence which satisfy P, then our algorithm TurboOptLift locates very quickly where property P occurs by chance on a sequence, and where it occurs as a result of a significant process. Under some conditions, the time complexity of TurboOptLift is O(n log n). We illustrate its use on the practical problem of locating approximate tandem repeats in DNA sequences. PMID- 10380203 TI - MDL and the statistical mechanics of protein potentials. AB - The combination of a wealth of structural data and impressive computational power provides detailed information pertaining to the structure and dynamics of biomacromolecules. A natural inclination is to incorporate this information into models to gain added predictive power on protein folding and stability. There has been considerable recent interest in developing "knowledge-based" potentials to describe internal interactions in proteins. In these approaches, probability distribution functions are inferred from existing knowledge. A common assumption has been the "quasi-chemical approximation" or "Boltzmann device". This method relates statistical mechanical probabilities to observed frequencies. The validity of this approach is discussed in detail from a statistical mechanics perspective. Because statistical mechanics is a form of statistical inference based on a lack of knowledge of the system, the "Boltzmann device" does not have a rigorous theoretical justification. In the present work, a statistical mechanics based on partial knowledge of the system is employed. This statistical mechanical scheme uses the minimum description length (MDL) of phase space as its main tool. With this approach, "knowledge-based" potentials can be derived in a rigorous fashion. In practical calculations, these potentials are best obtained using Bayesian inference methods similar to those used in image reconstruction. PMID- 10380204 TI - Using information theory to discover side chain rotamer classes: analysis of the effects of local backbone structure. AB - An understanding of the regularities in the side chain conformations of proteins and how these are related to local backbone structures is important for protein modeling and design. Previous work using regular secondary structures and regular divisions of the backbone dihedral angle data has shown that these rotamers are sensitive to the protein's local backbone conformation. In this preliminary study, we demonstrate a method for combining a more general backbone structure model with an objective clustering algorithm to investigate the effects of backbone structures on side chain rotamer classes and distributions. For the local structure classification, we use the Structural Building Blocks (SBB) categories, which represent all types of secondary structure, including regular structures, capping structures, and loops. For classification of side chain data, we use Minimum Message Length (MML) clustering from information theory. We show an example of how MML clustering on data classified by backbone SBBs can reveal different distributions of rotamer classes among the SBBs. Using these preliminary results, some of the characteristics of a rotamer library created using MML clustering on SBB dependent rotamer data are demonstrated. PMID- 10380205 TI - Improving the efficiency of a user-driven learning system with reconfigurable hardware. Application to DNA splicing. AB - This paper describes a new approach to problem solving by splitting up problem component parts between software and hardware. Our main idea arises from the combination of two previously published works. The first one proposed a conceptual environment of concept modelling in which the machine and the human expert interact. The second one reported an algorithm based on reconfigurable hardware system which outperforms any kind of previously published genetic data base scanning hardware or algorithms. Here we show how efficient the interaction between the machine and the expert is when the concept modelling is based on reconfigurable hardware system. Their cooperation is thus achieved with an real time interaction speed. The designed system has been partially applied to the recognition of primate splice junctions sites in genetic sequences. PMID- 10380206 TI - Optimizing Smith-Waterman alignments. AB - Mutual correlation between segments of DNA or protein sequences can be detected by Smith-Waterman local alignments. We present a statistical analysis of alignment of such sequences, based on a recent scaling theory. A new fidelity measure is introduced and shown to capture the significance of the local alignment, i.e., the extent to which the correlated subsequences are correctly identified. It is demonstrated how the fidelity may be optimized in the space of penalty parameters using only the alignment score data of a single sequence pair. PMID- 10380207 TI - Computing minimum description length for robust linear regression model selection. AB - A minimum description length (MDL) and stochastic complexity approach for model selection in robust linear regression is studied in this paper. Computational aspects and implementation of this approach to practical problems are the focuses of the study. Particularly, we provide both algorithms and a package of S language programs for computing the stochastic complexity and proceeding with the associated model selection. A simulation study is then presented for illustration and comparing the MDL approach with the commonly used AIC and BIC methods. Finally, an application is given to a physiological study of triathlon athletes. PMID- 10380208 TI - Topology selection in unrooted molecular phylogenetic tree by minimum model-based complexity method. AB - In reconstruction of phylogenetic trees from molecular data, it has been pointed out that multifurcate phylogenetic trees are difficult to be correctly reconstructed by the conventional methods like maximum likelihood method(ML). In order to resolve this problem, we have been engaged in developing a new phylogenetic tree reconstruction method, based on the minimum complexity principle widely used in the inductive inference. Our method, which we call "minimum model-based complexity (MBC) method", has been proved so far to be efficient in estimating multifurcate branching when the tree is described in the form of rooted one. In this study, we make further investigations about the efficiency of MBC method in estimating the multifurcation in unrooted phylogenetic trees. To do so, we conduct computer simulation in which the estimations by MBC method are compared with those by ML, AIC and statistical test approach. The results show that MBC method also provides good estimations even in the case of multifurcate unrooted trees and suggest that it could be generally used for reconstruction of phylogenetic tree having arbitrary multifurcations. PMID- 10380209 TI - Proteinmorphosis: a mechanical model for protein conformational changes. AB - Proteinmorphosis is a physically-based interactive modeling system for simulating large or small conformational changes of proteins and protein complexes. It takes advantage of the cross-linked one-dimensional nature of protein chains. The user can, based on her chemical knowledge, pull pairs of points (lying either on a single protein or on different molecules) together by specifying geometric distance constraints. The resulting conformation(s) of the molecule(s) of interest is computed by an efficient finite element formalism taking into account elasticity of the protein backbone, van der Waals repulsions, hydrogen bonds, salt bridges and the imposed distance constraints. The conformational change is computed incrementally and the result can be visualized as an animation; complete interactivity is provided to position and view the proteins as desired by the user. Physical properties of regions on the protein can also be chosen interactively. The conformational change of calmodulin upon peptide binding is examined as a first experiment. It is found that the result is satisfactory in reproducing the conformational change that follows on peptide binding. We use Proteinmorphosis to study the cooperative hemoglobin oxygen binding mechanism in a second, more sophisticated, experiment. Different modeling strategies are designed to understand the allosteric (cooperative) binding process in this system and the results are found to be consistent with existing hypotheses. PMID- 10380210 TI - Protalign: a 3-dimensional protein alignment assessment tool. AB - Protein fold recognition (sometimes called threading) is the prediction of a protein's 3-dimensional shape based on its similarity to a protein of known structure. Fold predictions are low resolution; that is, no effort is made to rotate the protein's component amino acid side chains into their correct spatial orientations. The goal is simply to recognize the protein family member that most closely resembles the target sequence of unknown structure and to create a sensible alignment of the target to the known structure (i.e., a structure sequence alignment). To facilitate this type of structure prediction, we have designed a low resolution molecular graphics tool. ProtAlign introduces the ability to interact with and edit alignments directly in the 3-dimensional structure as well as in the usual 2-dimensional layout. It also contains several functions and features to help the user assess areas within the alignment. ProtAlign implements an open pipe architecture to allow other programs to access its molecular graphics capabilities. In addition, it is capable of "driving" other programs. Because amino acid side chain orientation is not relevant in fold recognition, we represent amino acid residues as abstract shapes or glyphs much like Lego (tm) blocks and we borrow techniques from comparative flow visualization using streamlines to provide clean depictions of the entire protein model. By creating a low resolution representation of protein structure, we are able to at least double the amount of information on the screen. At the same time, we create a view that is not as busy as the corresponding representations using traditional high resolution visualization methods which show detailed atomic structure. This eliminates distracting and possibly misleading visual clutter resulting from the mapping of protein alignment information onto a high resolution display of the known structure. This molecular graphics program is implemented in Open GL to facilitate porting to other platforms. PMID- 10380211 TI - PROMUSE: a system for multi-media data presentation of protein structural alignments. AB - We present and evaluate PROMUSE: an integrated visualization/sonification system for analyzing pairwise protein structural alignments (superpositions of two protein structures in three-dimensional space). We also explore how the use of sound can enhance the perception and recognition of specific aspects of the local environment at given positions in the represented molecular structure. Sonification presents several opportunities to researchers. For those with visual impairment, data sonification can be a useful alternative to visualization. Sonification can further serve to improve understanding of information in several ways. One use for data sonification is in tasks such as background monitoring, in which case sounds can be used to indicate thresholding events. With PROMUSE, data represented visually may be enhanced or disambiguated by adding sound to the presentation. This aspect of data representation is particularly important for showing features that are difficult to represent visually, due to occlusion or other factors. Another feature of our system is that by representing some variables through sound and others visually, the amount of information that may be represented simultaneously is extended. Our tool aims to augment the power of data visualization rather than replace it. To maximize the utility of our sonifications to represent data, we employed musical voices and melodic components with unique characteristics. We also used sound effects such as panning a voice to the left or right speaker and changing its volume to maximize the individuality of the sonification elements. By making the sonification parameters distinct, we allow the user to focus on those portions of the sonification necessary to resolve possible ambiguities in the visual display. Sonifications of low level data such as raw protein or DNA sequences tend to sound random, and not very musical. We chose instead to sonify an analysis of data features, and thereby present a higher level view of the data. We also used brief melodic phrases rather than single notes in order to generate sounds that were more pleasing and musically idiomatic. To validate the utility of our system, we present the results of an experiment in which PROMUSE was used to test the use of sound as an aid for clarifying visual information. We also compare the overall effectiveness of visual versus aural information delivery. PMID- 10380212 TI - Developing protein documentaries and other multimedia presentations for molecular biology. AB - Computer-based multimedia technology for distance learning and research has come of age--the price point is acceptable, domain experts using off-the-shelf software can prepare compelling materials, and the material can be efficiently delivered via the Internet to a large audience. While not presenting any new scientific results, this paper outlines experiences with a variety of commercial and free software tools and the associated protocols we have used to prepare protein documentaries and other multimedia presentations relevant to molecular biology. A protein documentary is defined here as a description of the relationship between structure and function in a single protein or in a related family of proteins. A description using text and images which is further enhanced by the use of sound and interactive graphics. Examples of documentaries prepared to describe cAMP dependent protein kinase, the founding structural member of the protein kinase family for which there is now over 40 structures can be found at http://franklin.burnham-inst.org/rcsb. A variety of other prototype multimedia presentations for molecular biology described in this paper can be found at http://fraklin.burnham-inst.org. PMID- 10380213 TI - BioJAKE: a tool for the creation, visualization and manipulation of metabolic pathways. AB - The BioJAKE program has been created for the visualization, creation and manipulation of metabolic pathways. It has been designed to provide a familiar and easy-to-use interface while still allowing for the input and manipulation of complex and detailed metabolic data. In recognition of the detailed and diverse sources of data available across the Internet, it also provides a mechanism by which remote database queries can be stored and performed with respect to individual molecules within a pathway. This remote database access functionality is offered in addition to comprehensive local database creation, management and querying capability. The program has been developed in Java so as to provide for platform independence and maximum extendibility. PMID- 10380214 TI - Integrating computation and visualization for biomolecular analysis: an example using python and AVS. AB - One of the challenges in biocomputing is to enable the efficient use of a wide variety of fast-evolving computational methods to simulate, analyze, and understand the complex properties and interactions of molecular systems. Our laboratory investigates several areas including molecular visualization, protein ligand docking, protein-protein docking, molecular surfaces, and the derivation of phenomenological potentials. In this paper we present an approach based on the Python programming language to achieve a high level of integration between these different computational methods and our primary visualization system AVS. This approach removes many limitations of AVS while increasing dramatically the inter operability of our computational tools. Several examples are shown to illustrate how this approach enables a high level of integration and inter-operability between different tools, while retaining modularity and avoiding the creation of a large monolithic package that is difficult to extend and maintain. PMID- 10380215 TI - Field-based similarity forcing in energy minimization and molecular matching. AB - A new field-based similarity forcing procedure for matching conformationally flexible molecules is presented. The method extends earlier work on similarity matching of molecules based upon the program MIMIC, by directly coupling a similarity function to a molecular mechanics force field. In this way conformational energetics are fully accounted for in the similarity matching process. Simultaneous similarity/conformational searches can then be undertaken within a Monte Carlo or molecular dynamics framework. Here, a Monte Carlo approach is used to provide a simple example of two HIV-1 reverse transcriptase inhibitors, nevirapine and alpha APA, that illustrates the basic characteristics of the method and suggests areas for further investigation. PMID- 10380216 TI - Thermodynamics and kinetics of ligand-protein binding studied with the weighted histogram analysis method and simulated annealing. AB - The thermodynamics of ligand-protein molecular recognition is investigated by the energy landscape approach for two systems: methotrexate(MTX)--dihydrofolate reductase(DHFR) and biotin-streptavidin. The temperature-dependent binding free energy profile is determined using the weighted histogram analysis method. Two different force fields are employed in this study: a simplified model of ligand protein interactions and the AMBER force field with a soft core smoothing component, used to soften the repulsive part of the potential. The results of multiple docking simulations are rationalized from the shape of the binding free energy profile that characterizes the thermodynamics of the binding process. PMID- 10380217 TI - Empirical vs. "rational" methods of discovering new drugs. AB - Empirical and theoretical approaches to drug discovery have often been perceived as mutually exclusive. Our experience has rather demonstrated that they can be complementary. The structure-based approach to design of compound libraries is clearly helpful; however, testing large libraries continues to reveal unanticipated actives in many of our programs. A rationale for these observations is offered. PMID- 10380218 TI - Binary QSAR: a new method for the determination of quantitative structure activity relationships. AB - A new method (particularly suited to the analysis of High Throughput Screening data) is presented for the determination of quantitative structure activity relationships. The method, termed "Binary QSAR," accepts binary activity measurements (e.g., pass/fail or active/inactive) and molecular descriptor vectors as input. A Bayesian inference technique is used to predict whether or not a new compound will be active or inactive. Experiments were conducted on a data set of 1947 molecules. The results show that the method exhibits high accuracy and is robust to measurement errors. PMID- 10380219 TI - Ligand-receptor 3-D similarity studies using multiple 4-point pharmacophores. AB - A new method for 3-D similarity is presented based on the multiple potential 4 point 3-D pharmacophores expressed by ligands and complementary to receptors. These are calculated for ligands taking conformational flexibility into account, and for receptors through the use of complementary site-points. Through this common frame of reference both ligand-ligand and ligand-receptor similarity studies are possible. The application of the method to selectivity between different serine proteases (thrombin, factor Xa and trypsin) is discussed, and the need to use 4-point pharmacophores rather than 3-point pharmacophores is illustrated. A novel refinement to the potential pharmacophore method that uses a "special" feature to give a relative measure of similarity/diversity is also discussed. PMID- 10380220 TI - Two-dimensional reaction of biological molecules studied by Weighted-Ensemble Brownian dynamics. AB - Computer simulations offer critical insights into the reaction of biological macromolecules, especially when the molecular shapes are too complex to be amenable to analytical solution. In this work, the Weighted-Ensemble Brownian (WEB) Dynamics simulation algorithm is adapted to a reaction of two unlike biological molecules, with the interaction modeled by a two-parameter system: a spherical molecular depositing on a target region of an infinite cylinder with a periodic boundary conditions. The original algorithm of Huber and Kim is streamlined for this class of reactive models. The reaction rate constant is calculated as a function of relative sizes of the reactive to non-reactive regions of the cylindrical molecule. An analytical expression for the rate constant is also obtained from the solution of the diffusion equation for the special case of a constant-flux boundary condition. Good agreement between analytical and simulation results validates the applicability of WEB Dynamics to a reaction of molecules of complicated shape. On the other hand, the simple form of our analytical expression is useful as a testing case for other simulation and numerical techniques. PMID- 10380221 TI - Application of parameter optimization to molecular comparison problems. AB - Various bioinformatics comparison problems require optimizing several different properties simultaneously. Often linear objective functions combine the values for different properties of solution candidates into a single score to allow for multivariate optimization. In this context, an essential question is how each property should be weighted. Frequently, no apparent measure is available to serve as a model for the score. However, if preferences of certain solution candidates over others in a training set are available, the implied partial ordering may be used to best possibly adjust the weights. We apply different strategies to optimize the parameterization of empirical scoring functions used for two molecular comparison problems, protein threading and small molecule superposition. Using well established evaluation methods, it can be shown that the results of both comparison methods are significantly improved by systematically choosing appropriate weights for the scoring function contributions. PMID- 10380222 TI - Protein evolution and protein folding: non-functional conserved residues and their probable role. AB - It is shown that there are two types of conserved residues in evolutionary and functionally related proteins whose sequences have been well diverged in evolution. The first group consists of residues forming the active center, while the second (first established in this work) has nothing to do with function and therefore should be related to protein structure and/or protein folding. The lattter group consists of 4 residues in c-type cytochromes and 6 residues in globins. All these residues belong to alpha-helices and occupy positions (i, i + 4) or (i, i + 3), stabilizing one helical turn in some helices. These residues form an interface between the N- and C-terminal helices in c-type cytochromes and helices A, G, H in globins. These helical complexes form early in protein folding and are relatively stable in both equilibrium and kinetic folding intermediates. The attractive hypothesis is that these helices form folding nuclei in protein in the frame of the nucleation-growth mechanism of protein folding. PMID- 10380223 TI - A combined approach for ab initio construction of low resolution protein tertiary structures from sequence. AB - An approach to construct low resolution models of protein structure from sequence information using a combination of different methodologies is described. All possible compact self-avoiding C alpha conformations (approximately 10 million) of a small protein chain were exhaustively enumerated on a tetrahedral lattice. The best scoring 10,000 conformations were selected using a lattice-based scoring function. All-atom structures were then generated by fitting an off-lattice four state phi/psi model to the lattice conformations, using idealised helix and sheet values based on predicted secondary structure. The all-atom conformations were minimised using ENCAD and scored using a second hybrid scoring function. The best scoring 50, 100, and 500 conformations were input to a consensus-based distance geometry routine that used constraints from each the conformation sets and produced a single structure for each set (total of three). Secondary structures were again fitted to the three structures, and the resulting structures were minimised and scored. The lowest scoring conformation was taken to be the "correct" answer. The results of application of this method to twelve proteins are presented. PMID- 10380224 TI - Chaperonin function depends on structure and disorder in co-chaperonin mobile loops. AB - Co-chaperonins from diverse organisms exhibit mobile loops which fold into a beta hairpin conformation upon binding to the chaperonin. GroES, Gp31, and human Hsp10 mobile loops exhibit a preference for the beta hairpin conformation in the free co-chaperonins, and the conformational dynamics of the human Hsp10 mobile loop appear to be restricted by nascent hairpin formation. Backbone conformational entropy must weigh against binding of co-chaperonins to chaperonins, and thus the conformational preferences of the loops may strongly influence chaperonin-binding affinity. Indeed, subtle mutations in the loops change GroEL-binding affinity and cause defects in chaperonin function, and these defects can be suppressed by mutations in GroEL which compensate for the changes in affinity. The fact that high-affinity co-chaperonin binding impairs chaperonin function has implications for the mechanism of chaperonin-assisted protein folding. PMID- 10380225 TI - Similarity between the sequences of taxol-selected peptides and the disordered loop of the anti-apoptotic protein, Bcl-2. AB - The anti-cancer drug taxol is known to bind to and induce the polymerization of tubulin and has recently been shown to bind to the anti-apoptotic protein Bcl-2, but not to its homolog, Bcl-XL. Libraries of random peptides displayed on the surface of a bacteriophage were screened to select those exhibiting affinity for taxol. The sequences of these peptides were compared to sequences of proteins involved in mitosis and apoptosis. No significant similarities were detected between the sequences of tubulins and the taxol-selected peptides. However, a high level of similarity exists between the selected peptides and the disordered loop of Bcl-2. Conversely, there was little similarity between the sequences of the selected peptides and Bcl-XL. These results indicate that peptides displayed on the surface of a bacteriophage can mimic the ligand-binding behavior of a disordered protein loop and that comparison of the sequences of affinity-selected peptides with protein sequences can be predictive for ligand binding. PMID- 10380226 TI - Characterisation of side-chain conformational preferences in a biologically active but unfolded protein. AB - A combination of experimental NMR 3J alpha beta coupling constant measurements and theoretical predictions from a statistical model for a random coil have been used to characterise the conformations of amino acid side-chains in an unfolded fibronectin binding protein. The statistical model uses the distribution of torsion angles in a data base of native folded protein structures to provide a description of the torsion angle populations of each residue in a random coil. For all but three of the residues studied a close agreement is observed between the experimental 3J alpha beta data and the model predictions (correlation coefficient 0.90; RMSD 0.70 Hz). In these cases the populations about the chi 1 torsion angles in the conformational ensemble defining the fibronectin binding protein are well described by those present in the protein data base. For Phe 69, Asp 92 and Asp 105 however significant deviations are observed between the predictions and experimental data. Each of these side-chains is found to be involved in persistent non-random structural features arising from clustering of hydrophobic groups or interactions between charged side-chains. The analysis demonstrates the detailed insight that can be provided into conformationally disordered states by combining experimental and theoretical approaches. PMID- 10380227 TI - The locally denatured state of glutathione S-transferase A1-1: transition state analysis of ligand-dependent formation of the C-terminal helix. AB - On the basis of available x-ray structures, A-class glutathione S-transferases (GSTs) contain at their C-termini a short alpha-helix that provides a 'lid' over the active site in the presence of the reaction products, glutathione-conjugates. However, in the ligand-free enzyme this helix is disordered and crystallographically invisible. An aromatic cluster including Phe-10, Phe-220, and the catalytic Tyr-9 within the C-terminal strand control the order of this helix. Here, preliminary x-ray crystallographic analyses of the wild type and F220Y rGSTA1-1 in the presence of GSH are described. Also, a transition state analysis is presented for ligand-dependent formation of the helix, based on variable temperature stopped-flow fluorescence. Together, the results suggest that the ligand-dependent ordering of the C-terminal strand occurs with a transition state that is highly desolvated, but with few intramolecular hydrogen bonds or electrostatic interactions. However, substitutions at Phe-220 modulate the activation parameters through interactions with the side chain of Tyr-9. PMID- 10380228 TI - One sequence, four folds: transitions between an ensemble of metastable folds for the N-terminal domain of CD2. AB - Recombinant forms of the N-terminal domain of the cell adhesion receptor CD2 adopt a variety of olds by exchange of beta-sheets between adjacent polypeptide chains. Although these interdigitated forms are normally metastable, we have used site-directed mutagenesis to alter the kinetics of formation and relative stabilities of these states, leading to spontaneous formation of monomeric, dimeric, trimeric and tetrameric intertwined folded states. A characteristic feature of these fold-disorder-alternative fold transitions is the independence of each domain folding event, as deduced from kinetic analysis of folding data. Structures for fully interdigitated trimeric and tetrameric forms have been modelled, consistent with both the crystallographic and kinetic data. Although the biological role of these alternative folded states remains unclear, these structures form a remarkable demonstration of the fluidity of structure generated from a single polypeptide chain. PMID- 10380229 TI - The hydrophilic, protease-sensitive terminal domains of eucaryotic DNA topoisomerases have essential intracellular functions. AB - The amino-terminus of eucaryotic DNA topoisomerase I and the carboxy-terminus of eucaryotic DNA topoisomerase II contain sequences that are enriched in charged amino acid residues, hyper-sensitive to protease digestion, not required for the in vitro topoisomerase activities, able to tolerate insertion and deletion mutations, and thus may have a disordered structure. In an interesting contrast to the catalytically essential core domain, the sequences in these terminal hydrophilic domains are not conserved among the topoisomerases from different species. However, many lines of evidence, including those presented here, demonstrate that the topoisomerase tail domains have critical intracellular functions. The biological functions of the amino-terminus of topoisomerase I include the nuclear import and targeting to the transcriptionally active loci. The carboxy-terminus of topoisomerase II also contains the sequences necessary for nuclear localization and possibly sequences necessary for other critical functions. PMID- 10380230 TI - Recognition between disordered polypeptide chains from cleavage of an alpha/beta domain: self-versus non-self-association. AB - Advances in structural biology have provoked a re-evaluation of the biological significance of the disordered state of proteins. We believe that the rules that govern structure, stability and kinetics in the molecular recognition between disordered polypeptide chains can be elucidated by studying processes that couple association with folding. The reassembly of single domain proteins by fragment complementation provides an excellent opportunity to study them. Since almost the complete sequence is available, although not on a single chain, most of the complementary fragments are expected to reassemble. However, that happens not to be the case. We have chosen E. coli thioredoxin (Trx), a small, single alpha/beta domain protein, as a model system to study the effect of the site and number of cleavages on the reassembly of complementary fragments. We have shown at atomic detail the reassembly after cleavage of a loop (1-73, 74-108) and after cleavage of an alpha-helix (1-37, 38-108). Although both sets of fragments produce native like complexes, there are clear differences in the interface geometry, apparent stability of the folded state and mechanism of association/folding: (i) the apparent equilibrium dissociation constant for 1-37/38-108 complex (4 microM) is higher than the one for 1-73/74-108 complex (49 nM), (ii) the apparent rate constants of non-self-association are similar (about 10(3) M-1s-1), and (iii) only the 1-37 fragment self-associates under these experimental conditions. Here the competition between self- and non-self-association leads to an apparently less stable 1-37/38-108 complex. PMID- 10380231 TI - New therapeutic targets for rheumatoid arthritis. AB - New insights into the pathogenesis of rheumatoid arthritis (RA) and consequently new targets of therapy are covered in a broad overview fashion. Short-term significant beneficial effect on RA disease activity has been established in a small but rapidly growing number of double-blind placebo-controlled trials now including recombinant human IL-1 receptor antagonist, chimeric (mouse/human) monoclonal antibodies (mAb) against TNF alpha (cA2), humanised (human/mouse) anti TNF alpha mAb (CDP571) and recombinant human TNF-receptor-Fc fusion protein (TNFR:Fc). Placebo-controlled trials of anti-T cells agents such as chimeric anti CD4 mAb (cM-T412) and anti-CD5 immunoconjugate, did not demonstrate clinical benefit. A placebo-controlled study of the anti-T cell derived cytokine IL-2 (DAB486IL-2) showed only modes clinical improvement. Other anti-T cell approaches such as autologous T cell vaccination and induction of tolerance by oral type II collagen have been unsuccessful. The one controlled trial with an anti inflammatory cytokine, recombinant human IFN-gamma, showed modest clinical benefits. Controlled trials with IL-4 and IL-10 and with anti-adhesion molecules are awaited. PMID- 10380232 TI - Cost-effectiveness analysis of tropisetron vs. chlorpromazine-dexamethasone in the control of acute emesis induced by highly emetogenic chemotherapy in children. AB - OBJECTIVE: To perform a cost-effectiveness analysis (CEA) between a standard antiemetic regimen-chlorpromazine + dexamethasone (CPM-DEX)- and a 5-HT3 receptor antagonist-tropisetron (TROP)--in the control of acute emesis induced by highly emetogenic chemotherapy in children, considering two analytic perspectives: hospital and patients. METHODS: The CEA was performed by constructing a decision tree, for both analytic perspectives, of the possible outcomes of treatment with TROP (single 0.2 mg/kg i.v.) or CPM (5-15 mg i.v. infusion for 3 doses) plus DEX (2 mg/m2 i.v. bolus i.v. x2). The patients were stratified by age in two groups (2-12 and 13-17). To estimate the probability of each endpoint at the decision tree we have taken as a base a trial developed in the Department of Pediatrics. Direct medical cost of primary therapy, failure, complications and side effects were included in the cost calculations. RESULTS: From patients' analytic perspective, TROP was more cost-effective than CPM-DEX for both groups of patients. Discrepancy between both analytic perspectives in 13-17 year-old patient's group was resolved in favour of the option chosen from the patients' analytic perspective (TROP). Sensitivity analysis showed the reliability of the results. CONCLUSIONS: 1. TROP was more cost-effective than CPM-DEX. 2. Taking into account the patients' analytic perspective is essential when we compare antiemetics pharmacoeconomically. 3. It seems necessary to increase the effectiveness of TROP in pediatric patients receiving highly emetogenic chemotherapy with strategies such as the addition of a steroid. PMID- 10380233 TI - The importance that community pharmacists in Malta place on the introduction of pharmaceutical care. AB - The results from a study to assess the importance Maltese pharmacists placed on various aspects of pharmaceutical care and their willingness to provide such care are reported. A modified version of the Behavioural Pharmaceutical Care Scale (BPCS) questionnaire (consisting of three dimensions and 14 domains) was mailed to the 198 privately owned community pharmacies in Malta. A total of 99 questionnaires were returned following two reminder telephone calls. Pharmacists were asked to score the importance of each pharmaceutical care activity contained in the modified BPCS on a 6 point Likert scale ranging from 0 to 5. The overall score for the questionnaire, which illustrated the importance pharmacists attributed to various aspects of pharmaceutical care, ranged from 90 to 170 with a mean score of 134.8. There was little difference recorded between the scores for the three dimensions. The Referral and Consultation domain recorded a slightly higher score than the other two domains. Younger pharmacists obtained significantly higher scores (p < 0.05; Kruskal-Wallis test) in the Verification of Patient Understanding domain. Approximately 72% of respondents indicated that they were willing to provide pharmaceutical care, but remarked that a number of issues e.g. reimbursement, qualified support staff, GP-pharmacist co-operation, had to be addressed. A series of strategic steps are needed to help pharmacists resolve these issues before pharmaceutical care programmes could be offered by Maltese community pharmacists. PMID- 10380234 TI - Rational pharmacotherapy in The Netherlands: formulary management in Dutch hospitals. AB - A survey regarding the management of rational pharmacotherapy was conducted among all Dutch general hospitals in 1998. The response was 99% (n = 120). The presence of a drugs and therapeutics committee and antibiotic policies in Dutch general hospitals appears independent of hospital characteristics. However, formulary agreements and treatment guidelines are less likely to be present in hospitals that employ only 1 pharmacist or those served by community pharmacies. More than half of the hospitals claim to have restrictive formulary agreements. Large hospitals, hospitals in the eastern and southern provinces and those served by hospital pharmacies more often tend to have restrictive agreements compared to small hospitals, hospitals in the northern, central, and western provinces, and those served by community pharmacies. Various methods to impose restriction and ensure formulary compliance are mentioned. It must be noted that hospitals tend to operate rather solely regarding the large number of different formularies. Surprisingly just a small majority of pharmacists evaluates formulary agreements positively as a management tool. Many drawbacks appear to be present. The results of this survey indicate that in the future Dutch hospitals will favour disease management (treatment guidelines) over drug management (formulary agreements) in the management of rational pharmacotherapy and that information technology will be used to influence clinicians' prescribing behaviour. PMID- 10380235 TI - Prescription data as a tool in pharmacotherapy audit (I). General considerations. AB - This article discusses how prescription data that are available from pharmacies can be used as a tool to support the improvement of prescribing behaviour. For the optimal use of prescription data in pharmacotherapeutic audit, the specific aim of the audit meeting must be clear. Next, appropriate measures (quality of drug choice, volume, cost) have to be selected for the presentation and discussion of prescribing. For several aims of audit meetings, suggestions for adequate measures are given. For each stage of behavioural change the article describes why and how prescription data can provide a useful contribution. Finally, recommendations are given regarding prerequisites for the optimal use of prescription data. Important factors are that the prescription data should sufficiently cover the physicians' patient population, information regarding the extent of repeat prescribing is needed, and the prescribing of drugs with multiple indications can cause difficulties with the interpretation of the graphs. With this information in mind, feedback on prescribing can be tailored to the audit groups' needs and targeted to the intervention that is pursued. PMID- 10380236 TI - Prescription data as a tool in pharmacotherapy audit (II). The development of an instrument. AB - In many countries, prescription data are used as an instrument to provide feedback on prescribing. This article describes the development and implementation of a computer program as a tool for pharmacotherapy audit meetings and it illustrates the possibilities of such a program. The program was developed to support the optimisation of prescribing in the various stages of behavioural change. It enables pharmacists to make sophisticated overviews of prescribing by physicians in their audit group and furthermore, pharmacists can conduct drug utilisation studies or pharmaco-epidemiological studies in their own patient population. Forty percent of Dutch pharmacies have purchased the computer program since its introduction; it was distributed together with a technical handbook and a course book, which elaborated on the types of graphs available and their interpretation. An educational course was developed regarding the optimal selection and interpretation of graphs. Examples of prescription data as a tool to support various aims of audit meetings are given. The program is acknowledged as a welcome contribution to pharmacotherapy audit and to the pharmacists' advisory role in pharmacotherapy. PMID- 10380237 TI - Flurbiprofen, S(+), eyedrops: formulation, enantiomeric assay, shelf-life and pharmacology. AB - Aphakic cystoid macula edema, occurring after cataract extraction is ascribed to trauma-induced production of intra-ocular prostaglandins. Sufficient experimental and clinical evidence supports the use of prostaglandin synthesis inhibitors to countervail this clinical condition. The active S(+)-enantiomer of flurbiprofen, a prostaglandin synthesis inhibitor, has been formulated into a stereoselective, ballast free eyedrop solution in a concentration of 0.015%. Analysis by capillary zone electrophoresis shows shelf-life stability up to four years at room temperature of this enantiomer. The inhibitory effect on the synthesis of prostaglandins as measured on a homogenate bovine iris/ciliary body, remained unaffected during a shelf-life period of three years. PMID- 10380238 TI - Drugs up in smoke: a study of caseated drugs in Sweden. AB - DESIGN: Cross-sectional survey. SETTING: The county of Malmohus, with 817,000 inhabitants, in the far south of Sweden. All drugs handed in for destruction to the 65 pharmacies during one week in March 1996 were analysed. RESULTS: 92% of the packages were prescription drugs for human use, 7% were over-the-counter drugs, and 1% were for veterinary use. Slightly less than half (48%) had expired when they were handed in for destruction. 36% were unbroken when returned and another 18% of the packages were nearly full. A comparison between the drugs sent in for destruction and the drugs sold in the county gave a ratio of 0.030. Antineoplastic and immunosuppressive drugs, drugs for the respiratory system, antiparasitic products, and cardiovascular drugs were returned to a greater extent than other types of drugs. Drugs for the genito-urinary tract, sex hormones, and drugs for the alimentary tract were returned to a lesser extent. Extrapolated to a whole year the value of caseated drugs was estimated to 60 SEK (5.83 Br,) per person. The value of unbroken packages was 20 SEK (1.94 Br,) per person per year. CONCLUSION: Although not all of the drugs handed in for destruction could have been unnecessarily prescribed or obtained by the patient, a more cautious approach to prescribing of drugs would likely yield significant savings. PMID- 10380239 TI - Treatment of epilepsy in the elderly. AB - Management of epilepsy in the elderly involves many challenges, including the presence of concomitant diseases, polypharmacy and changes in body physiology. Age-related changes in pharmacokinetics and pharmacodynamics have to be taken into account in order to avoid potentially severe adverse drug reactions in elderly people. The present study reviews the most commonly used antiepileptic drugs (AEDs) in the elderly. Because some AEDs may induce the metabolism of other agents and reduce the effectiveness of several drugs, the physicians have to carefully evaluate concomitant drugs being administered. Moreover, the main problems appear to be when beginning therapy, the first choice drug, the appropriate dosage and pharmacologic compliance. Elderly patients must be screened for hepatic and renal functions before beginning a treatment with an AED, carefully interviewed to reduce complaints for drug side-effects which may negatively influence compliance and monitored for total and free blood levels. Besides the 'classic' AEDs, such as phenytoin, phenobarbital, carbamazepine, valproic acid, primidone and benzodiazepines, the review shows the possible advantages of new AEDs, such as felbamate, gabapentin, lamotrigine, oxcarbazepine and gamma-vinyl-GABA, which may be used in the elderly too for their good tolerability. A careful control of drug assumption is requested in the elderly, especially when it is difficult to achieve seizure control. PMID- 10380240 TI - Muscarinic acetylcholine receptors in the hippocampus, neocortex and amygdala: a review of immunocytochemical localization in relation to learning and memory. AB - Immunocytochemical mapping studies employing the extensively used monoclonal anti muscarinic acetylcholine receptor (mAChR) antibody M35 are reviewed. We focus on three neuronal muscarinic cholinoceptive substrates, which are target regions of the cholinergic basal forebrain system intimately involved in cognitive functions: the hippocampus; neocortex; and amygdala. The distribution and neurochemistry of mAChR-immunoreactive cells as well as behaviorally induced alterations in mAChR-immunoreactivity (ir) are described in detail. M35+ neurons are viewed as cells actively engaged in neuronal functions in which the cholinergic system is typically involved. Phosphorylation and subsequent internalization of muscarinic receptors determine the immunocytochemical outcome, and hence M35 as a tool to visualize muscarinic receptors is less suitable for detection of the entire pool of mAChRs in the central nervous system (CNS). Instead, M35 is sensitive to and capable of detecting alterations in the physiological condition of muscarinic receptors. Therefore, M35 is an excellent tool to localize alterations in cellular cholinoceptivity in the CNS. M35-ir is not only determined by acetylcholine (ACh), but by any substance that changes the phosphorylation/internalization state of the mAChR. An important consequence of this proposition is that other neurotransmitters than ACh (especially glutamate) can regulate M35-ir and the cholinoceptive state of a neuron, and hence the functional properties of a neuron. One of the primary objectives of this review is to provide a synthesis of our data and literature data on mAChR-ir. We propose a hypothesis for the role of muscarinic receptors in learning and memory in terms of modulation between learning and recall states of brain areas at the postsynaptic level as studied by way of immunocytochemistry employing the monoclonal antibody M35. PMID- 10380241 TI - Edwin Bancroft Henderson: physical educator, civil rights activist, and chronicler of African American athletes. PMID- 10380242 TI - A 30-s chair-stand test as a measure of lower body strength in community-residing older adults. AB - Measuring lower body strength is critical in evaluating the functional performance of older adults. The purpose of this study was to assess the test retest reliability and the criterion-related and construct validity of a 30-s chair stand as a measure of lower body strength in adults over the age of 60 years. Seventy-six community-dwelling older adults (M age = 70.5 years) volunteered to participate in the study, which involved performing two 30-s chair stand tests and two maximum leg-press tests, each conducted on separate days 2-5 days apart. Test-retest intraclass correlations of .84 for men and .92 for women, utilizing one-way analysis of variance procedures appropriate for a single trial, together with a nonsignificant change in scores from Day 1 testing to Day 2, indicate that the 30-s chair stand has good stability reliability. A moderately high correlation between chair-stand performance and maximum weight-adjusted leg press performance for both men and women (r = .78 and .71, respectively) supports the criterion-related validity of the chair stand as a measure of lower body strength. Construct (or discriminant) validity of the chair stand was demonstrated by the test's ability to detect differences between various age and physical activity level groups. As expected, chair-stand performance decreased significantly across age groups in decades--from the 60s to the 70s to the 80s (p < .01) and was significantly lower for low-active participants than for high active participants (p < .0001). It was concluded that the 30-s chair stand provides a reasonably reliable and valid indicator of lower body strength in generally active, community-dwelling older adults. PMID- 10380243 TI - The learning advantages of an external focus of attention in golf. AB - This study examined whether the learning advantages of an external focus of attention relative to an internal focus, as demonstrated by Wulf, Hoss, and Prinz (1998), would also be found for a sport skill under field-like conditions. Participants (9 women, 13 men; age range: 21-29 years) without experience in golf were required to practice pitch shots. The practice phase consisted of 80 practice trials. One group was instructed to focus on the arm swing (internal focus), whereas another group was instructed to focus on the club swing (external focus). One day after practice, a retention test of 30 trials without instructions was performed. The external-focus condition was more effective for performance during both practice and retention. PMID- 10380245 TI - Techniques of body composition assessment: a review of laboratory and field methods. AB - Body composition is one of the major health-related components of fitness. Thus, it is important for health and fitness professionals to have a general understanding of the most commonly used techniques for assessing body composition. This review presents the developmental background and underlying principles and theory of four laboratory (hydrodensitometry, air displacement plethysmography, isotope dilution, and dual-energy x-ray absorptiometry) and four field (bioelectrical impedance analysis, near-infrared interactance, skinfolds, and anthropometry) methods of body composition assessment. In addition to a description of the methods, the validity, and reliability, strengths, and limitations of each measurement tool are examined. Highlights of the laboratory methods include the relatively new Bod Pod air displacement device, which is a promising assessment tool more convenient than hydrodensitometry but still lacking substantial validity testing and the ability of dual-energy x-ray absorptiometry to measure regional composition making it an attractive method for clinicians. Advancements in segmental and multifrequency bioelectrical impedance for compartmental analysis have enhanced the value of this field method, but research continues to show that commercially available near-infrared interactance units are invalid. With this knowledge, the clinician and researcher should be able to make an informed decision regarding the most appropriate measurement device for their body composition assessments. PMID- 10380244 TI - Effects of health-related physical education on academic achievement: project SPARK. AB - The effects of a 2-year health-related school physical education program on standardized academic achievement scores was assessed in 759 children who completed Metropolitan Achievement Tests before and after the program. Schools were randomly assigned to condition: (a) Specialists taught the Sports, Play, and Active Recreation for Kids curriculum; (b) classroom teachers were trained to implement the curriculum; and (c) controls continued their usual programs. The Trained Teacher condition was superior to Control on Language, Reading, and Basic Battery. The Specialist condition was superior to Control on Reading, but inferior on Language. Despite devoting twice as many minutes per week to physical education as Controls, the health-related physical education program did not interfere with academic achievement. Health-related physical education may have favorable effects on students' academic achievement. PMID- 10380246 TI - Effect of body composition on oxygen uptake during treadmill exercise: body builders versus weight-matched men. AB - Oxygen uptake (VO2) during treadmill exercise is directly related to the speed and grade, as well as the participant's body weight. To determine whether body composition also affects VO2 (ml.kg-1.min-1) during exercise, we studied 14 male body builders (M weight = 99 kg, SD = 7; M height = 180 cm, SD = 8; M body fat = 8%, SD = 3; M fat free mass = 91 kg, SD = 7) and 14 weight-matched men (M weight = 99 kg, SD = 9; M height = 179 cm, SD = 5; M body fat = 24%, SD = 5; M fat free mass = 73 kg, SD = 9). Percentage of body fat, t(13) = 8.185, p < .0001, and fat free mass, t(13) = 5.723, p < .0001, were significantly different between groups. VO2 was measured by respiratory gas analysis at rest and during three different submaximal workrates while walking on the treadmill without using the handrails for support. VO2 was significantly greater for the lean, highly muscular men at rest: 5.6 +/- 1 vs. 4.0 +/- 1 ml.kg-1.min-1, F(1, 26) = 21.185, p < .001; Stage 1: 1.7 mph/10%, 18.5 +/- 2 vs. 16.1 +/- 2 ml.kg-1.min-1, F(1, 26) = 6.002, p < .05; Stage 2: 2.5 mph/12%, 26.6 +/- 3 vs. 23.1 +/- 2 ml.kg-1.min-1, F(1, 26) = 7.991, p < .01; and Stage 3:3.4 mph/14%, 39.3 +/- 5 vs. 33.5 +/- 5 ml.kg-1.min-1, F(1, 26) = 7.682, p < .01, body builders versus weight-matched men, respectively. However, net VO2 (i.e., exercise VO2 - rest VO2) was not significantly different between the two groups at any of the matched exercise stages. The findings from this study indicate that VO2 during weight-bearing exercise performed at the same submaximal workrate is higher for male body builders compared to that measured in weight-matched men and that which is predicted by standard equations. These observed differences in exercise VO2 appear to be due to the higher resting VO2 in highly muscular participants. PMID- 10380247 TI - Sport injury and illness: elite skiers describe their experiences. AB - Injury and illness pose significant physical and psychological challenges for competitive athletes. The present study examined the psychological processes associated with injury and illness among elite skiers. Twelve members of the Canadian Alpine Ski Team who had recovered from a serious sport injury or debilitating illness were interviewed for the study. Inductive analyses of interview transcripts revealed that the experience of sport injury and athlete illness spanned three distinct phases, namely, the injury-illness phase, the rehabilitation-recovery phase, and the return to full activity phase. In addition, it was found that each phase was marked by a series of events that caused the skiers varying degrees of distress. This article traces the skiers' psychological journey from injury and illness through recovery, with an emphasis on factors contributing to stress and the strategies used to manage stress. PMID- 10380248 TI - Four-year changes in college athletes' ethical value choices in sports situations. AB - Positive values for fairness in competition are supposed to undergird the behavior of athletes engaged in sport. Whether athletes' values actually develop over 4 years in a college that emphasizes character development is the focus of this study. Athletes' (N = 631) use of deontological ethics (Hahm, Beller & Stoll, 1989) in 21 sports value dilemmas were evaluated. At entrance, as well as near graduation, intercollegiate athletes' value scores were lower than intramural athletes' scores. Both groups' scores declined while they were in college. Individual-sport athletes had higher scores than team-sport athletes but manifested a greater decline over 4 years. The findings are consistent with other studies that show decreases in "sportsmanship orientation" and an increase in "professional" attitudes associated with participation in sport. PMID- 10380249 TI - Individual differences, perceived task characteristics, and preferences for teaching and coaching. AB - One hundred ninety-two undergraduate physical education students participated in this study of the influence of individual differences, group, and task factors on respondents' preferences for teaching or coaching. The individual difference factors were gender, managerial potential (Gough, 1984), and sex role attributes (Spence & Helmreich, 1986). The group and task differences were measured by a specially developed scale that was factor analyzed to yield six factors: job status, job significance, job variety and control, job identity, ease of discipline, ease of motivation. Subgroup analyses showed that men preferred more to coach than women did, and women preferred more to teach than men did. Men and women perceived greater job variety in coaching and greater control in teaching. The genders did not differ in managerial motivation or in any of the other perceived characteristics of the teaching and coaching roles. Finally, preference for teaching or coaching was influenced by gender, perceived ease of motivating students and athletes, and perceived job variety. These results were discussed in relation to existing literature, and future research directions are identified. PMID- 10380250 TI - Planning and executing fast movement sequences: a comparison of speech and handwriting. PMID- 10380251 TI - Aerobic dance and physical self-perceptions in female adolescents: some implications for physical education. PMID- 10380252 TI - Does participation in a structured high-intensity exercise program influence daily physical activity patterns in older adults? PMID- 10380253 TI - The effect of temporal and informational constraints on one-handed catching performance. PMID- 10380254 TI - Is the WHO analgesic ladder for cancer pain management appropriate for rheumatology patients? PMID- 10380255 TI - Long-term cyclosporin continuation rates in rheumatoid arthritis patients. AB - OBJECTIVE: To evaluate the continuation rate of cyclosporin therapy in rheumatoid arthritis patients followed for at least three years. METHODS: Retrospective medical chart review of rheumatoid arthritis patients on cyclosporin. Treatment efficacy was assessed based on a visual analog scale pain score, Ritchie's articular index, and Lee's functional index. Nonparametric Kaplan-Meier survival curves were used to evaluate continuation rates. RESULTS: 24 cyclosporin-treated patients with a mean age of 58 years and a mean disease duration of ten years were included in the study; 87% had received three second-line drugs prior to cyclosporin. Mean cyclosporin treatment duration was 28 months (range, 1-103 months). Overall cyclosporin continuation rates were 75% after four months and 50% after 36 months. Toxicity and inefficacy caused 33% and 13% of cyclosporin discontinuations, respectively. CONCLUSION: The continuation rate of cyclosporin was satisfactory and similar to that reported for other second-line drugs. PMID- 10380256 TI - Relationship of rheumatoid factor isotype levels with joint lesions detected by magnetic resonance imaging in early rheumatoid arthritis. AB - OBJECTIVE: To evaluate the relationship between rheumatoid factor isotypes and articular damage detected by magnetic resonance imaging and plain radiography in early rheumatoid arthritis. METHODS: 20 consecutive patients with early active rheumatoid arthritis underwent determinations of serum IgM, IgA, and IgG rheumatoid factors by enzyme-linked immunosorbent assay (ELISA). Plain radiographs of the hands and wrists were obtained, and the wrist, metacarpophalangeal joints, and proximal interphalangeal (PIP) joints on the more severely affected side were investigated by magnetic resonance imaging before and after gadolinium-DTPA injection. RESULTS: IgM, IgA, and IgG rheumatoid factors were found in 13 (65%), 13 (65%), and 15 (75%) of patients, respectively. Sera from five patients (25%) contained no detectable rheumatoid factor isotypes. Correlations were found among the levels of the three rheumatoid factor isotypes. Levels of IgA, IgG, and IgM rheumatoid factor were significantly higher in patients with than without erosions on magnetic resonance imaging scans. No such difference was found when patients with and without erosions on plain radiographs were compared. Magnetic resonance imaging detected soft tissue lesions more frequently than plain radiography. Magnetic resonance imaging was also more likely than plain radiography to show bone erosions and bone cysts, but this difference was not statistically significant. CONCLUSIONS: Quantitative rheumatoid factor isotype assays and magnetic resonance imaging evaluation of erosions of the hand and wrist may be useful for investigating patients with early rheumatoid arthritis. PMID- 10380257 TI - Primary synovial osteochondromatosis of the hand and wrist. Report of a series of 21 cases and literature review. AB - OBJECTIVES: To define the characteristics of synovial osteochondromatosis of the hand and wrist. PATIENTS AND METHODS: Retrospective study of 21 patients, including 11 with intraarticular and 10 with tenosynovial disease. Cases secondary to degenerative joint disease were excluded. Surgery consisted in removal of the osteocartilaginous bodies and of the adjacent synovial membrane. Mean follow-up was seven years (range, three to 18 years). The relevant literature was reviewed in part. RESULTS: Recurrence was seen in four patients and was multiple in two of these four. Most recurrences occurred within five to ten years after surgery. All four patients with recurrences had intraarticular disease. No cases of malignant transformation were seen. The characteristics of synovial osteochondromatosis at the hand and wrist are reviewed. CONCLUSION: Detailed preoperative investigations including computed arthrotomography and magnetic resonance imaging should be performed to increase the likelihood of complete excision. PMID- 10380258 TI - Reasons for rheumatology department admission in 125 patients with disk-related sciatica. AB - OBJECTIVES: To identify medical and nonmedical reasons for admission of disk related sciatica patients. PATIENTS AND METHODS: 125 patients were evaluated prospectively using a 25-items questionnaire, including seven items on medical reasons, four on psychological reasons, four on work-related reasons, six on social and family reasons, and four on miscellaneous reasons. RESULTS: Severe nerve root pain (34%), motor loss (17%), atypical clinical manifestations (13%), severe low back pain (8%), and/or sphincter dysfunction (4%) were recorded in only 55% of patients, and only 16% had at least two of these reasons. A minority of patients were admitted to avoid premature surgery (13%) or to try one more conservative approach prior to surgery (15%). Seventy-five per cent of patients reported at least one of the psychological reasons listed in the questionnaire (irritability/fatigue, 66%; anxiety, 42%; depression, 26%; panic disorder, 21%), 50% reported at least one work-related reason (workaholism, 21%; job offer, 16%; self-employed, 14%; fear of losing their job, 11%), 66% reported at least one social or family reason (living alone, 34%; one or more dependents younger than seven years of age, 32%; too many demands from household members, 22%; one or more dependents older than seven years of age, 8%; need to care for another person, 9%; important upcoming family or personal event, 6%), and 26% reported at least one miscellaneous reason (firm belief that sciatica can be cured only by inhospital treatment, 10%; desire to put pressure on the employer or on an expert, 7% and 6%, respectively; admission via the emergency room without prior medical advice, 6%). CONCLUSION: In France, the reason for admission of patients with disk-related sciatica is frequently a mixture of physical, psychological, and social problems, with only 55% of patients having a symptom requiring inhospital management. PMID- 10380259 TI - Morphine in nonmalignant joint pain. Physiological rationale, value, and limitations. PMID- 10380260 TI - Rheumatology in Italy at the threshold of the year 2000. PMID- 10380261 TI - A case of Schnitzler's syndrome with nodular regenerative hyperplasia of the liver. AB - Schnitzler's syndrome is a rare condition of urticaria, macroglobulinemia, and sclerotic bone lesions. We report a case in a 70-year-old man in whom inflammatory polyarthralgia was followed by a nonpruritic urticarial eruption with a moderate decline in general health. Laboratory tests showed inflammation and a modest isolated peak of monoclonal IgM kappa. There was no evidence of Waldenstrom macroglobulinemia. Schnitzler's syndrome was considered. However, an ultrasound scan of the abdomen done because of mild gamma-glutamyl-transferase elevation disclosed multiple hepatic lesions. The liver histology showed incipient nodular regenerative hyperplasia. Only about 30 cases of Schnitzler's syndrome have been reported since the seminal description in 1972. Hepatic involvement was a common but nonspecific finding, and we found no cases with nodular regenerative hyperplasia. However, this abnormality is often found in patients with autoimmune or hematological disorders. The pathogenesis of Schnitzler's syndrome remains unknown, but the possibility of progression to a hematological malignancy requires prolonged follow-up. PMID- 10380262 TI - Neuropathic arthropathy: a forgotten diagnosis? Two recent cases involving the hip. AB - Two cases of neuropathic arthropathy of the hip are reported. One was the first manifestation of tabes dorsalis in a 74-year-old man, whereas the second occurred in a 47-year-old woman with a history of spina bifida and L2-L5 epidural lipoma. Radiographic joint destruction occurred within five and three months, respectively. The main clinical and radiological features of neuropathic arthropathy are reviewed, and diagnostic pitfalls are discussed. There are no specific laboratory tests or histologic findings. This now rare condition should be routinely considered in patients with severe joint destruction contrasting with minimal pain. The reasons for the decision to use trochanteric-iliac coaptation in one of our patients and a wait-and-see approach in the other are explained. The literature is reviewed. Ten cases treated by joint replacement have been reported. However, neither joint replacement nor arthrodesis seem capable of restoring satisfactory hip function. PMID- 10380264 TI - Cystic rheumatoid arthritis with Felty's syndrome and ankylosing spondylitis. AB - A 63-year-old man with strictly axial ankylosing spondylitis since the age of 28 years had a seven-year history of cystic seronegative rheumatoid arthritis with Felty's syndrome. Cysts were present in the hands, feet, wrists, shoulders, hips, one elbow, and one knee. There was no evidence of juxtaarticular demineralization, joint space loss, erosions, or joint destruction. Rheumatoid pannus was demonstrated within the cysts, particularly at the hip, ruling out cystic hip disease due to ankylosing spondylitis. HLA typing demonstrated the B27 and DR4 haplotypes. HLA B27 may be associated with a worse prognosis of rheumatoid hip involvement. PMID- 10380263 TI - Hydatid cyst of the sacrum. Report of a case. AB - An unusual case of hydatid cyst of the sacrum revealed by low back pain and sciatica in a 16-year-old is reported. Computed tomography and a surgical biopsy provided the diagnosis. The outcome was favorable one year after mebendazole therapy initiation. PMID- 10380265 TI - Sarcoid tenosynovitis. Report of a case. PMID- 10380266 TI - Hemarthrosis of the ankle revealing an aneurysm of the anterior tibial artery. PMID- 10380267 TI - Meniscus replacement--is it really necessary? PMID- 10380268 TI - Meniscus transplantation--consensus declaration. PMID- 10380269 TI - Function of the normal meniscus and consequences of meniscal resection. AB - The principal functions of the meniscus are load transmission and shock absorption, based on the meniscal collagen architecture, the biochemical fluid composition, and the proteoglucan-collagen meshwork. The mobile menisci transmit 50-90% of load over the knee joint, depending on knee flexion angle, femoral translation and rotation. The meniscus contributes to knee joint proprioception and probably also to joint stability. Late consequences of total and partial meniscectomy are radiographic osteoarthritis, with a varying percentage of these patients having symptoms. Malalignment, concomitant articular cartilage lesions, and ligament instability are absolute risk factors, while age, lateral compartment, and continued sport activity are relative risk factors. Acute reinsertion of meniscal tears in the red-red or red-white zones can be performed successfully by arthroscopic technique. Also in chronic tears stable healing can be expected in most cases, if the scar tissue is resected. PMID- 10380270 TI - Meniscal substitutes--animal experience. AB - Animal studies have shown that meniscus allografts and tendon autografts generally heal to the capsule, are revascularized and repopulated with host cells. In animals, neither meniscal allografts nor tendon or fat autografts gain the properties of a normal meniscus. Meniscus allografts and tendon autografts are promising as both seem to offer some protection to the cartilage of the tibial plateau. There is no evidence that meniscal transplantation can prevent cartilage degenerative changes, and the long-term effect of meniscal transplantation on articular cartilage remains unknown. Whether cellular repopulation of the meniscal allograft is sufficient to restore its biomechanical properties is unknown. Collagen scaffolds and tissue engineered grafts are still under investigation, showing promising results especially for the former. Viable meniscal allografts should be implanted within 1 to 2 weeks after harvesting, as the production of proteoglycans decreases after 2 weeks. PMID- 10380271 TI - Meniscal substitutes--human experience. AB - A number of clinical series have described the effect of meniscus allograft replacement in humans. The general indication has been disabling pain following loss of a meniscus in a skeletally mature individual. Overall, healing of the graft to the capsule occurs in up to 80% of all transplants. Revascularization and cell repopulation is found in all grafts but is highly variable. The risk for graft failure seems to be greater with irradiated grafts and in patients with grade III or IV osteoarthritic changes. In most series, patients experienced a decrease in pain and an increase in activity level postoperatively. In many series, concominant surgery (cruciate ligament reconstruction or osteotomy) had been performed. Meniscus replacement with frozen or cryopreserved allografts seems to give the most promising short-term results in patients with post meniscectomy pain. Controlled, randomized prospective studies are needed to confirm a long-term benefit and better define transplantation indications. Viable meniscus allografts seem to survive transplantation, as donor cells were found in the graft after 2 years. Clinically, pain was reduced and activity increased following transplantation, but after 4 years some of these gains were lost. There was no correlation between postoperative findings on MRI and clinical outcome. Meniscal replacement with a quadriceps tendon autograft in humans resulted in pain reduction, but at second-look arthroscopy, only 2 of 9 tendon autografts looked like a meniscus. Six were in position but still looked like tendons. Total medial meniscus replacement by quadriceps tendon autrograft is still an experimental procedure. There is no proof at present that meniscal substitutes (meniscus allografts or tendon autografts) in humans can protect the hyaline cartilage of the knee from the degeneration, following loss of a meniscus. There is some evidence in animal experiments that under circumstances not yet exactly known, a meniscus substitute can have a protective effect on articular cartilage. Three factors have been identified that prevent proper meniscal function: poor fixation of the meniscal horns, no contact of the graft with the articulating surfaces under load and incorrect positioning of the horns. Meniscal allograft transplantation sensitizes humoral and cell mediated immune systems. Bone plugs attached to meniscal allograft tissue may increase cell surface antigenicity. Deep freezing and especially freeze drying of meniscal tissue decreases host immunogenicity. Cryopreservation maintains the content of donor HLA encoded antigens and is likely more sensitizing to the host. The clinical importance of immune responses to meniscal allografts is not known, but it has not been shown to result in graft failure or rejection. Prospective studies are needed. PMID- 10380272 TI - Harvest and conservation of meniscal allografts. AB - Meniscal allografts can be harvested and preserved in different ways. A technique for harvesting and conserving viable allografts is described. The meniscus should be harvested within 12 h after the start of ischaemia. The ideal time for implantation of the viable graft is 10 to 14 days after harvest, in which time a recipient should be identified. Frozen grafts are avital. Cryopreserved grafts are not avital, as 10-30% of the fibrochondrocytes are preserved. Frozen and cryopreserved grafts can be stored for a long time and if a graft bank is established, grafts are always readily available. There is no proven advantage of one graft type over others. PMID- 10380273 TI - Meniscus transplantation: preoperative planning. AB - Precise preoperative planning is mandatory to obtain good results of meniscus transplantation. Any pathology in addition to the missing meniscus should be detected, including malalignment, and ligament instability. A full status of the changes in the cartilage should be made. It is unknown how precisely the donor meniscus needs to fit the size of the original meniscus to allow healing and regeneration. The effect of incongruency and wrong insertion of the meniscal horns has been investigated in an animal experiment. The incongruous grafts did not entirely avoid chondromalacia after meniscectomy, but provided some chondroprotective effect. A wrong bony insertion of the meniscal horns leads to even more serious chondral damage than meniscectomy. PMID- 10380274 TI - Meniscal regeneration or meniscal transplantation? AB - Owing to the initial necrosis to which any freely transplanted biological graft is subjected, meniscus transplantation is based on similar principles to meniscal regeneration. Both methods rely on repopulation of extrinsic cells of the graft. In the former procedure a biological matrix (meniscus, tendon, fatpad) is used as graft (scaffold), whereas in meniscal regeneration commercially available resorbable or non-resorbable scaffolds are implanted. However, the cellular (re)population and (re)vitalization process is deleterious rather than beneficial for the function of the graft as the remodelling of the tissue leads to shrinkage and narrowing of the implant. In addition, improper fixation and subsequent elongation of the anterior and posterior bony attachments leads to peripheral graft dislocation, loss of the load distribution function, and subsequently to cartilage degeneration. Hence, meniscus transplantation or regeneration faces two major problems: 1) remodelling of graft to inferior tissue properties after allograft transplantation, or lacking establishment of normal tissue properties after use of biological matrixes other than the meniscus (fatpad, tendon), or commercially available matrixes; 2) improper fixation with elongation of the anterior and posterior attachments. Furthermore, use of allografts incorporates the risk for disease transmission. Today we are unable to control these problems, and therefore the concept of meniscal replacement does not work yet. Further research is necessary to control remodelling and improve fixation to make this procedure a successful one in the future. PMID- 10380275 TI - Arthroscopic and open surgical techniques for meniscus replacement--meniscal allograft transplantation and tendon autograft transplantation. AB - Open, arthroscopically assisted and arthroscopic methods for lateral and medial meniscus allograft transplantation with bone plug fixation are described. An open technique for medial and lateral meniscus transplantation without bone plug fixation, as well as an open technique for autograft quadriceps tendon replacement of the medial meniscus are described. PMID- 10380276 TI - Postoperative follow-up and rehabilitation after meniscus replacement. AB - Even though basic scientific knowledge about the meniscal loading pattern may advocate restrictive rehabilitation after meniscus repair, experience and one controlled study favour accelerated rehabilitation. The current protocols for rehabilitation after meniscal substitution follow personal experience with parameters such as pain, effusion, locking, and gait pattern used as clinical guidance. Controlled clinical studies on rehabilitation should be encouraged. An example of a rehabilitation protocol is given. As meniscus replacement is a new treatment option, it is essential to document details about the graft, the knee status at operation and the surgical procedure. The goal of a postoperative follow-up is to control quality, to measure patient satisfaction and to show whether meniscus replacement is beneficial in relation to the natural history of meniscectomy cases. There is a need for a standard follow-up evaluation of patients after meniscus replacement, and the development of a meniscus transplantation score including subjective and objective data is suggested. PMID- 10380277 TI - Future research in meniscal replacement. AB - To study the remodelling process after meniscus replacement and to learn how to control it will be a key topic for future research. Not enough is known about the importance of precision in meniscal fixation and how to make the insertions as strong as in the normal meniscus. Methods to measure load-distribution, mechanical properties of the graft and status of the cartilage should be developed. A number of different auto- and allografts have been shown to heal and revascularize, but whether the grafts are functioning is not known. Meniscal scaffolds, which can be used for replacement of partial meniscal loss, need to be tested. Scaffolds and tissue engineering will be the subject of intensive research in the future. PMID- 10380278 TI - Indications for meniscal transplantation. Who and how many need a meniscus substitute? A personal view. PMID- 10380279 TI - Meniscus transplantation should not be combined with correction of instability/deformity/cartilage defects. A personal view. PMID- 10380280 TI - It is necessary to anchor the meniscal transplants with bone plugs? A mini battle. PMID- 10380281 TI - Temporomandibular disorders and mandibular function in relation to Class II malocclusion and orthodontic treatment. A controlled, prospective and longitudinal study. AB - The relationship between orthodontic treatment and symptoms and signs of temporomandibular disorders (TMD) was studied prospectively and longitudinally in 65 adolescent girls with Class II malocclusion. The subjects received orthodontic fixed appliance treatment with the straight-wire technique combined with or without extractions and were examined for symptoms and signs of TMD before, during, after, and finally one year post-treatment. Both symptoms and signs of TMD showed considerable fluctuations over the three-year period within the individuals. The general tendency was a decreased prevalence of symptoms of TMD over the three years. The prevalence of pain on mandibular movement and tenderness to palpation of the masticatory muscles was significantly less common during and after orthodontic treatment than before. Clinically registered TMJ clicking increased slightly over the three year period. One orthodontic treatment effect when normalizing Class II malocclusions with fixed appliances was a decreased prevalence of functional occlusal interferences. We concluded that the orthodontic treatment either with or without tooth extractions did not increase the risk for TMD or worsen pre-existing signs of TMD. Subjects with Class II malocclusion and pre-treatment signs of TMD of muscular origin seemed rather to benefit functionally from orthodontic treatment in a three-year perspective. PMID- 10380282 TI - On oral disease, illness and impairment among 50-year-olds in two Swedish counties. AB - There were three general aims of the thesis. To investigate 1) the fulfillment of the judiciary Swedish goal of good dental care on equal conditions in an adult population born 1942 in two Swedish counties in 1992. Both selfrated oral health and clinical observations indicated good oral health in the study population. Full social equality of dental care was not attained, but the social gradient was modest. 2) To find models for risk prediction on population level for oral health, expressed as number of remaining teeth, as caries and as periodontitis. Relatively efficient such models were constructed, with the lowest efficiency for periodontitis indicators. Oral health and disease were found to be qualitatively different. Still, use of tobacco was a consistent risk factor for both poor health and presence of disease. 3) To investigate if questionnaires can be used to monitor oral health, disease and illness. It was found that this is the case on population level, but also that questionnaire surveys cannot wholly replace clinical studies, especially regarding periodontal disease. For number of remaining teeth and frequency of removable dentures, the questionnaire methodology is appropriate for monitoring in this type of population, which has been utilized by setting the present study as a baseline for ongoing longitudinal studies relying on survey methodology. PMID- 10380283 TI - [The paradox of genome size and the problem of redundant DNA]. AB - Data on the relationship between genome size and phenotypic traits of animals and plants are reviewed. Different concepts explaining accumulation of redundant (noncoding) DNA in genomes of eukaryotes are discussed: regulatory (repressory), neutralistic (permissive), skeletal, bodyguard, and buffering, respectively. The relationship between genome size and metabolic rate is presumably the primary one. Possible molecular mechanisms responsible for this relationship are considered. It is concluded that the problem of redundant DNA (genome size paradox) is hardly to solve by studies on molecular level only, since the genome size is a parameter related to molecular, cellular and organismal levels. Redundant DNA seems to be of ecophysiological significance, and therefore the interdisciplinary field dealing with genome size paradox can be called "genome cytoecology". PMID- 10380284 TI - [Electron-microscopic and immunohistochemical study of the reorganization of the microtubule system in granular cells of frog urinary bladder after water transport induction]. AB - Structural changes in organization of the microtubule system in granular cells of frog urinary bladder after water transport induction by vasopressin were studied by methods of electron microscopy and immunocytochemistry. It is shown that in steady-state conditions microtubules form a wide network equally distributed in the whole cytoplasm of granular cells. After vasopressin action, the amount of microtubules increases in the apical region of the cytoplasm. A predominant orientation of microtubules, perpendicular to the apical membrane direction, appears. A structural association of microtubules with specific granules and large vacuoles was observed. A supposition is advanced about association of the described microtubule system reorganization with the activation of vectorial intracellular transport occurring after transepithelial water transport induction. PMID- 10380285 TI - [The stimulating effect of destabilase, a component of Hirudo medicinalis salivary gland secretion, on sensory neuron neurite growth in organotypic culture]. AB - The effect of destabilase, a component of Hirudo medicinalis salivary gland secret, was investigated in organotypic tissue culture of dorsal root ganglia (DRG) of 10-11-day old chick embryos. Native destabilase in concentrations 0.01 and 0.05 ng/ml was active, inducing a more intensive neurite growth in DRG that in the control. The stabilizing activity of destabilase was lost following reverse-phase chromatography. Neurite-stimulating effects of the drug "pyjavit" is due presumably to neurite-stimulating activity of destabilase. PMID- 10380286 TI - [Role of topoisomerase II in the response of mammalian cells to the action of ionizing radiation. I. Change in the mitotic cell cycle of Chinese hamster CHO K1, irradiated with x-rays, after the action of novobiocin at a high concentration]. AB - Flow cytometry was used to study cell cycle recovery in X-irradiated Chinese hamster cells after action of novobiocin, an inhibitor of topoisomerase II. A prolonged treatment with 1 mM novobiocin (20-30 h) of intact cells results in the G2 + M delay. Novobiocin treatment of 5 Gy-irradiated cells results in a slight delay in cell exit from G1 into S phase and in a much longer G2-delay if compared with X-irradiated cells. These data allow to suggest an involvement of topoisomerase II in cell response to ionizing radiation. PMID- 10380287 TI - [Structure of chromatin and chromosomes in preparations of interphase nucleus derivatives, prepared by removal of the nucleuar envelopes. II. Structure of chromatin and associations of chromosomes in stretched amembranous nuclei and mitotic figures]. AB - Preparations of surface stretched amembranous nuclei and mitotic figures were used for revealing the high order nuclear and chromosomal structures. The preparations were obtained by dropping amembraneous nuclei and mitotic figures suspension in methanol-glacial acetic acid mixture (3:1) on wetted superclean slides. Amembraneous nuclei and mitotic figures were isolated from intact murine and human cells (lines L1210, SK-UT-1B, PHA-stimulated lymphocytes) by means of their 1-5 min prefixational capillary pipetting with freshly prepared 0.018-0.06% Triton X-100 solution in the conditional cultural medium. Stretched amembraneous nuclei and mitotic figures had no features of induced chromatin dispersion and compaction. Stretched interphase amembraneous nuclei showed spatially separated individual structures (thin chromatin fibres, nucleoli, intranuclear bodies), polymorphous pattern of perinucleolar chromatin aggregation and episodically expressed beaded thick chromatin fibres and a chromocenter. The chromomeric pattern of the spread chromosomes of mitotic figures was quite similar but hardly identical with that of G-banding. The stretched prometaphase mitotic figures in all tested cell types always contained loose "residual" nucleoli looking like typical prophase nucleoli as concerns their shape and number per cell (mitotic figure). The majority of chromosomes of stretched mitotic figures and of prophase amembraneous nuclei were attached to the nucleolar material. All tested cell lines showed almost the same variation in number of nucleolus-attached chromosomes, per both prophase amembraneous nucleus and prometaphase mitotic figure. Some chromosomes of stretched mitotic figures were colocated with "residual" nucleoli and looked shortened and strongly condensed. Other chromosomes, locally associated with "residual" nucleoli, were straight and oriented radially to these. Mutual chromosomal arrangements in mitotic cells on smears and in stretched mitotic figures were analogous. Equatorial plates from PBS-washed SK-UT-1B cells displayed a better stretching capacity than those from untreated cells. In the former case metaphase chromosomes were seen more uniformly stretched and well identified after GTG-banding procedure. The number of interchromosomal (mainly telomere-telomeric and telomere-centromeric) connections per stretched mitotic figure (or per stretched prophase amembraneous nucleus) was minimum in late prometaphase, maximum in prophase and early prometaphase, and intermediate in metaphase. The obtained data are discussed in terms of topology and longitudinal heterogeneity of mitotic chromosomes. PMID- 10380288 TI - [Laser spatial scanning in flow cytometry]. AB - The main flow laser cytometry principles, based on the elastic light scattering, spheres of its applications, problems of its realization and utilization in the immunological investigations and diagnostics are analysed. The experimental model of a flow cytometer with laser probing beam space scanning, originally proposed by the authors, is described. The apparatus was tested by polystyrene latex spheres and biological objects. The experiments showed that the achieved sensitivity was enough to register red blood cells, their complexes and bacterial cells. PMID- 10380289 TI - Fetal assessment: time to reassess. PMID- 10380290 TI - Early fetal gender determination. PMID- 10380291 TI - The sonographic identification of fetal gender from 11 to 14 weeks of gestation. AB - OBJECTIVE: To determine the feasibility of correctly identifying fetal gender from 11 to 14 weeks' gestation. METHODS: A prospective cross-sectional study in a university Department of Obstetrics and Gynaecology, London. A total of 524 women from an unselected population underwent a detailed assessment of fetal anatomy at 11-14 weeks of gestation (confirmed by crown-rump length) by means of transabdominal sonography, and transvaginal sonography (26%) when necessary. Fetal gender was identified in the transverse and sagittal planes, and was confirmed at birth. RESULTS: The overall success of correctly assigning fetal gender increased with gestational age from 46% to 75%, 79% and 90% at 11, 12, 13 and 14 weeks, respectively. The ability of the operator to assign fetal gender significantly improved with increasing gestational age (p < 0.0001), being 59%, 87%, 92% and 98% at 11, 12, 13 and 14 weeks, respectively. The accuracy of correctly identifying fetal gender when attempted did not change with gestational age. Fetal gender or the performance of the scan by different operators did not affect the results. CONCLUSION: Whilst the accuracy of sonographic determination of fetal gender at 11-14 weeks is good, it still falls significantly short of invasive karyotyping tests. PMID- 10380292 TI - First-trimester determination of fetal gender by ultrasound. AB - OBJECTIVE: To assess the accuracy of fetal sex determination at 11-14 weeks of gestation. METHODS: Fetal gender assessment by ultrasound was prospectively carried out in 172 singleton pregnancies at 11-14 weeks of gestation immediately before chorionic villus sampling for karyotyping. The genital region was examined in a midsagittal plane and the fetal gender was assigned as male if the angle of the genital tubercle to a horizontal line through the lumbosacral skin surface was greater than 30 degrees and female when the genital tubercle was parallel or convergent (less than 30 degrees) to the horizontal line. RESULTS: The accuracy of sex determination increased with gestation from 70.3% at 11 weeks, to 98.7% at 12 weeks and 100% at 13 weeks. In the male fetuses, there was a significant increase in the angle of the genital tubercle from the horizontal with crown-rump length. Male fetuses were wrongly assigned as female in 56% of cases at 11 weeks, 3% at 12 weeks and 0% at 13 weeks. In contrast, only 5% of the female fetuses at 11 weeks were incorrectly assigned as male and this false-positive rate was 0% at 12 and 13 weeks. CONCLUSION: The clinical value of determination of fetal sex by ultrasound is in deciding whether to carry out prenatal invasive testing in pregnancies at risk of sex-linked genetic abnormalities, because invasive testing would be necessary only in pregnancies with male fetuses. Our results suggest that a final decision on invasive testing for sex-linked conditions should be undertaken only after 12 weeks of gestation. PMID- 10380293 TI - Biometrical threshold of biparietal diameter for certain fetal sex assignment by ultrasound. AB - OBJECTIVES: The aim of this study was to establish the biometric threshold of biparietal diameter (BPD), assumed to be an independent variable of gestational age, at which 100% accuracy in the assessment of fetal sex by ultrasonography is achievable. METHODS: Transvaginal and/or transabdominal sonography was used for detecting the 'sagittal sign' as a marker of fetal sex in 385 fetuses with BPD between 18 and 29 mm. The results of ultrasound examination were compared with sex at birth or with karyotype obtained from amniotic fluid cells or chorionic villus sampling. RESULTS: Fetal sex assignment was feasible in 337 of 385 cases (87.5%). Of the 312 fetuses with known fetal sex outcome, 164 were males and 148 were females. An accuracy rate of 100% was achieved when a BPD of > or = 23 mm was obtained. CONCLUSION: This study provides important information about the earliest stage of fetal development, expressed in terms of BPD, at which a diagnosis of fetal sex can be made with 100% accuracy. PMID- 10380294 TI - Fetal heart rate and umbilical artery velocity variability in pregnancies complicated by insulin-dependent diabetes mellitus. AB - OBJECTIVES: To examine the variability in fetal heart rate and absolute flow velocity, which are possible hemodynamic markers of cardiovascular homeostasis in pregnancies complicated by diabetes mellitus. METHODS: Doppler studies of umbilical artery velocity waveforms were performed at 12-21 weeks of gestation in 16 women with well-controlled type I (insulin-dependent) diabetes mellitus. From umbilical artery velocity waveforms of at least 13 s in duration, we determined absolute values and beat-to-beat variability for fetal heart rate and umbilical artery flow velocities and compared these findings with normal controls matched for gestational age. RESULTS: Fetuses of diabetic women displayed increased fetal heart rate variability and umbilical artery peak systolic velocity. Fetal heart rate, umbilical artery time-averaged velocity and variability in umbilical artery flow velocity were not essentially different between the two groups. CONCLUSION: Fetal heart rate variability and umbilical artery peak systolic velocity may be markers for fetal cardiovascular homeostasis in pregnancies complicated by insulin-dependent diabetes mellitus. PMID- 10380295 TI - Maternal cardiolipin, beta 2-glycoprotein-I and prothrombin antibody expression in high-risk pregnancies with bilateral abnormal uterine artery Doppler waveforms. AB - OBJECTIVE: To compare the frequency of maternal serum antiphospholipid antibodies (to cardiolipin, beta 2-glycoprotein I and prothrombin) in pregnancies presenting with bilateral abnormal uterine artery Doppler waveforms. DESIGN: Retrospective analysis of stored serum. SUBJECTS: Cases comprised 47 singleton pregnancies with bilateral abnormal uterine artery Doppler waveforms at 24 weeks of gestation, followed from 20 weeks, and controls were 100 healthy pregnancies with normal uterine artery Doppler waveforms. METHODS: Ultrasound examination utilized a 5 MHz curvilinear transabdominal transducer with pulsed and color Doppler facilities. Antiphospholipid antibodies were analyzed by ELISA methodology, and reference ranges were established using the geometric mean +/- 2 SD of healthy non-pregnant adults. Human chorionic gonadotropin (hCG) levels were obtained from patient notes. RESULTS: Anticardiolipin antibodies were detected in 11 (23%) of the cases (IgG, n = 7; IgM, n = 6) compared with ten (10%) of the controls (p < 0.05). Low titer anticardiolipin IgG (range, 5.5-35.3; median, 6.3 GPL units) and anticardiolipin IgM (range, 3.4-14.7; median, 5.3 MPL units) were detected in cases. Amongst the cases, adverse perinatal outcomes were more common in the presence of raised levels of anticardiolipin antibodies. Anti-beta 2-glycoprotein I IgG was not detected in any of the cases. Antiprothrombin IgG was not detected, but antiprothrombin IgM occurred in 10.6% of cases compared with 2% of controls. CONCLUSIONS: Women with persistent bilateral abnormal uterine artery. Doppler waveforms in mid-gestation were more likely to express raised levels of anticardiolipin antibodies than healthy controls with normal uteroplacental perfusion. Anticardiolipin antibodies without anti-beta 2-glycoprotein I binding may be involved in the pathogenesis of uteroplacental ischemia in a proportion of high-risk pregnancies. PMID- 10380296 TI - Sonographic evaluation of antepartum development of fetal gastric motility. AB - OBJECTIVE: Little is known about the development of fetal gastric motility and emptying. The aim of this study was to evaluate the development of gastric motility sonographically in the human fetus. METHODS: The motility and peristalsis of the fetal stomach were sonographically studied in 76 normal fetuses at 12-39 weeks of gestation. Fetal gastric motility was assessed by videotaping real-time ultrasonic images of the stomach for periods of 60 or more minutes. RESULTS: Gastric peristalsis appeared as early as 14 weeks of gestation, and was detected in all fetuses by 23 weeks. The frequency of peristaltic waves was constant, and was 2.2-3 times per minute at 14-39 weeks of gestation. The onset of fetal gastric peristalsis was sporadic and the period with no peristaltic waves was dominant before 24 weeks of gestation. Fetal gastric peristalsis increased and consolidated into long-term clusters from 24 weeks of gestation. The mean duration of peristalsis increased from 4.1 +/- 1.2 min (n = 6) at 20-23 weeks to 14.1 +/- 3.2 min (n = 14) at 32-35 weeks of gestation, and remained constant thereafter. CONCLUSIONS: Fetal gastric motility was quantified and its development during pregnancy was assessed in this study. There was a critical point of development at around 24-25 weeks of gestation when grouped peristalsis was observed in all fetuses. PMID- 10380297 TI - Magnetic resonance imaging supplements ultrasonographic imaging of the posterior fossa, pharynx and neck in malformed fetuses. AB - OBJECTIVE: The objective of this study was to compare antepartum ultrasonography and magnetic resonance imaging (MRI) in the diagnosis and exclusion of malformations of the fetal neck, pharynx, skull base and posterior fossa in late pregnancy. MATERIALS AND METHODS: The study involved 26 women and 27 fetuses with ultrasonographically or clinically suspected abnormalities of the fetal neck, pharynx or central nervous system (CNS). Findings obtained by ultrasound were compared with those obtained by MRI (1.5 T) in the last trimester. RESULTS: In cases with CNS malformation (n = 19), MRI provided additional information on the anatomy of the foramen magnum and posterior fossa in nine cases (47%). When antepartum ultrasonography indicated malformation of the soft tissues of the neck or pharynx (n = 8), MRI provided additional information on diagnosis or exclusion of the abnormality in six cases (75%). The imaging capacity of the anatomy of the naso-, oro- and hypopharynx, trachea, esophagus and cervical skin outlines was better with MRI. CONCLUSIONS: MRI proved to be a valuable supplementary method to ultrasound in obtaining accurate information from the fetal neck, pharynx and posterior fossa, particularly when acoustic shadowing by bony structures or adjacent malformation impaired the quality of the ultrasonographic examination. PMID- 10380298 TI - Fetal lumbar spine volumetry by three-dimensional ultrasound. AB - OBJECTIVES: To evaluate three-dimensional sonographic volume measurements of the thoracolumbar spine from 16 to 25 weeks of gestation in the normally developing fetus. DESIGN: Prospective cross-sectional study. SUBJECTS: The study included 103 women between 16 and 25 weeks of gestation. They were enrolled at the time of their anomaly scan. None of the pregnancies was associated with structural anomalies and in each case fetal size was deemed appropriate for gestational age. METHODS: Three-dimensional volume calculation of the 1st and 5th lumbar vertebral body, the 12th thoracic vertebral body and the whole lumbar spine was performed, using the perpendicular frontal, sagittal and transverse planes. The lumbar spine length was determined. RESULTS: A statistically significant increase in all measurements was found with advancing gestational age. CONCLUSIONS: Three dimensional ultrasonography allows volume calculation of the fetal spine. Our study provides preliminary data about volume changes in the developing fetal spine between 16 and 25 weeks' gestation. PMID- 10380299 TI - Screening efficacy of the subcutaneous tissue width/femur length ratio for fetal macrosomia in the non-diabetic pregnancy. AB - BACKGROUND: Antenatal weight estimations have limited sensitivity and specificity for the detection of macrosomia. The objective of our study was to examine the screening efficacy of the subcutaneous tissue width/femur length ratio for the intrapartum detection of fetal macrosomia in a non-diabetic population at term. STUDY DESIGN: Intrapartum sonographic measurements were performed in 178 well dated gravidas at 37-41 weeks' gestation with negative glucose tolerance screens. The biparietal diameter, femur length (FL), abdominal circumference and subcutaneous tissue width of the thigh (SCT) were determined. Subsequently, predictions for macrosomia (actual birth weights above the 90th centile) were made using varying cut-off points of the examined parameters or estimated fetal weights. RESULTS: Macrosomia occurred in 27 newborns (15.1%). The SCT/FL ratio was independent of gestational age (r = -0.017). Maternal age, gravidity, parity, gestational age and the ratio of male-to-female infants were similar in pregnancies resulting in appropriate-for-gestational-age and macrosomic infants (NS). There was no difference in the SCT/FL ratio between these groups (p = 0.067; 99% power to detect 2 standard deviation differences). Comparison of screening efficacy by the univariate z score for the area under receiver operating characteristic (ROC) curves (theta) revealed that the abdominal circumference had the best sensitivity-specificity trade-off (theta = 0.8843; p < 0.0001 for comparison with SCT/FL ROC curve), followed by weight estimations based on the Hadlock formula (theta = 0.8773; p < 0.0005), the Shepard formula (theta = 0.8606; p < 0.0001), subcutaneous tissue thickness alone (theta = 0.6872; p < 0.01) and the SCT/FL ratio (theta = 0.6303). CONCLUSIONS: We conclude that the SCT/FL ratio is a poor sonographic predictor of fetal macrosomia in the non-diabetic pregnancy and does not improve fetal weight estimations by conventional sonographic parameters. PMID- 10380300 TI - Transvaginal sonographic ovarian findings in a random sample of women 25-40 years old. AB - OBJECTIVE: To investigate the occurrence rate of adnexal lesions in premenopausal women. METHODS: A random sample of women 25-40 years old was invited to undergo a transvaginal ultrasound examination, and 335 women were examined. The criteria used to define an adnexal lesion were either a cystic lesion with its largest diameter of at least 25 mm within the pelvic region, or the appearance of solid parts in any lesion regardless of size. RESULTS: Adnexal lesions were found in 26/335 cases, (7.8%) (95% confidence interval (CI), +/- 2.9%). The occurrence rate of ovarian cysts was 22/335 (6.6%) (95% CI, +/- 2.7%). There were no differences between the women with or without ovarian cysts related to age, smoking habits, parity or body mass index. At follow-up 3 months later, 18 of the 22 (82%) cysts had disappeared (95% CI, +/- 16%). Women using progesterone contraception (either oral contraception or an intrauterine device with levonorgestrel) had a significantly higher relative risk of 2.7 (95% CI, 1.1-6.9) of functional cysts as compared to women with natural cycles. Polycystic ovaries were found in 10.2% (95% CI, +/- 4.2%) of the women not using any hormonal contraception. The mean volumes of the polycystic ovaries were significantly larger compared to those in natural cycles. CONCLUSION: Adnexal lesions are common in asymptomatic women in the age group 25-40 years, but four out of five ovarian cysts disappeared spontaneously after 3 months. The ultrasound appearance of the cyst, the woman's family history and her own feelings must be considered if a persisting cyst is to be surgically removed or followed by repeated transvaginal ultrasound. PMID- 10380301 TI - Prenatal diagnosis of fetal adrenal masses: differentiation between hemorrhage and solid tumor by color Doppler sonography. AB - We present four cases of fetal adrenal masses detected by routine prenatal ultrasound, of which three were adrenal hemorrhages and one was a fetal neuroblastoma. The differential diagnoses of fetal adrenal mass include adrenal hemorrhage, neuroblastoma, adrenal and cortical renal cysts, pulmonary sequestrations, duplication of the renoureteral system and Beckwith-Wiedemann syndrome. These can pose a diagnostic and therapeutic dilemma. A systematic comparison of the pre- and postnatal sonographic features including color Doppler imaging may help to differentiate adrenal hemorrhage from neuroblastoma prenatally. PMID- 10380302 TI - First-trimester ultrasound diagnosis of holoprosencephaly: three case reports. AB - We present three cases of fetal holoprosencephaly diagnosed by transabdominal and transvaginal ultrasound examinations at 10 and 13 weeks' gestation. The diagnosis was based on two sonographic criteria: first, the intracranial finding of a single ventricle with a cerebral mantle and no visible midline structures but fusion of the thalami and corpus striatum; and, second, facial abnormalities, including hypotelorism. The ultrasound findings were confirmed by embryoscopy before abortion in one case and by pathological examination after abortion in two cases. Chromosome study of the three fetuses showed trisomy 18, triploidy and mosaic 18p deletion and duplication. PMID- 10380303 TI - Prenatal diagnosis of alobar holoprosencephaly at 10 weeks of gestation. AB - Alobar holoprosencephaly is an intracranial abnormality characterized by failure of proper cleavage of the prosencephalon, accompanied by incomplete midfacial development. The prenatal sonographic diagnosis of alobar holoprosencephaly was first described in 1984; however, there have been only two reports of alobar holoprosencephaly diagnosed in the first trimester. We report a case of alobar holoprosencephaly diagnosed at 10 weeks of gestation. PMID- 10380304 TI - In utero diagnosis of cardiac hemangioma. AB - Fetal cardiac hemangioma is rarely diagnosed prenatally. We present here a fetus with such a tumor diagnosed at 28 weeks' gestation. With the use of fetal echocardiography, a mixed echogenic mass protruding outward from the right atrial wall was observed. Moderate amounts of pericardial effusion were also found. Although no apparent blood flow signal was detected in the mass, fetal echocardiography showed signs suggestive of a hemangioma. Differential diagnosis, management and prognosis are discussed. PMID- 10380305 TI - Sonographic features of fetal trisomy 18 at 13 and 14 weeks: four case reports. AB - Fetal trisomy 18 is the second most common multiple malformation syndrome. We present four cases of trisomy 18 with multiple sonographic abnormalities at 13 and 14 weeks of gestation. These cases demonstrated that fetal hand deformities can be a tell-tale sign of trisomy 18 with or without increased nuchal translucency at this gestation. PMID- 10380307 TI - A case of anaphylactic reaction caused by exposure to a latex probe cover in transvaginal ultrasound scanning. PMID- 10380306 TI - Gastroschisis associated with bladder evisceration complicated by hydronephrosis presenting antenatally. AB - We report here a case of gastroschisis associated with bladder evisceration and complicated by rapidly developing hydronephrosis diagnosed antenatally. The timing of delivery was determined by the hydronephrosis, associated bowel dilatation and polyhydramnios. The case highlights the need for continuing ultrasonographic surveillance of fetuses with gastroschisis to identify further associated complications which were hitherto absent but whose presence may influence the timing of delivery and neonatal care. PMID- 10380308 TI - Primary uterine hydatid cyst and the wall-sign criteria. PMID- 10380309 TI - Menstrual history: authors' reply. PMID- 10380310 TI - Ultrasound, tamoxifen and endometrial carcinoma. PMID- 10380311 TI - Sonographic images of a lost intrauterine device. PMID- 10380312 TI - Importance of nuchal translucency measurement in multifetal pregnancy reduction. PMID- 10380314 TI - Three-dimensional ultrasonography of congenital ichthyosis. PMID- 10380313 TI - Ultrasound imaging of pseudomyxoma peritonei with numerous vesicles in ascitic fluid. PMID- 10380315 TI - A mixed approach and a distribution-free multiple imputation technique for the estimation of a multivariate probit model with missing values. AB - In the present paper a mixed generalized estimating/pseudo-score equations (GEPSE) approach together with a distribution-free multiple imputation technique is proposed for the estimation of regression and correlation structure parameters of multivariate probit models with missing values for an ordered categorical time invariant variable. Furthermore, a generalization of the squared trace correlation (RT2) for multivariate probit models, denoted by pseudo-RT2, is proposed. A simulation study was conducted, simulating a probit model with an equicorrelation structure in the errors of an underlying regression model and using two different missing mechanisms. For a low 'true' correlation the difference between the GEPSE, a generalized estimating equations (GEE) and a maximum likelihood (ML) estimator were negligible. For a high 'true' correlation the GEPSE estimator turned out to be more efficient than the GEE and very efficient relative to the ML estimator. Furthermore, the pseudo-RT2 was close to RT2 of the underlying linear model. The mixed approach is illustrated using a psychiatric data set of depressive in-patients. The results of this analysis suggest that the depression score at discharge from a psychiatric hospital and the occurrence of stressful life events seem to increase the probability of having an episode of major depression within a one-year interval after discharge. Furthermore, the correlation structure points to short-time effects on having or not having a depressive episode, not accounted for in the systematic part of the regression model. PMID- 10380316 TI - Measuring effect size: a non-parametric analogue of omega 2. AB - When comparing two groups of subjects, one of the many measures of effect size is omega 2. This paper suggests a non-parametric analogue of omega 2 based on the following point of view. Given an observation from one of two groups, but not knowing whether it came from the first or second group, how certain can we be that the observation came from the first group? This is in contrast to omega 2 where, given that an observation came from a specific group, say the first group, how much does this reduce our uncertainty about the dependent variable? One problem with omega 2 is that it is not robust--it is a function of the variances- so it can be misleading for reasons reviewed in the paper. Four estimators of the proposed measure of effect size are described and compared in a simulation study. Contrary to what was expected, the .632 bootstrap estimator performed best in terms of bias and mean squared error. PMID- 10380317 TI - Bayesian analysis of square ordinal-ordinal tables. AB - We analyse square contingency tables with ordered categories. Assuming that the observed ordinal categorical variables are manifestations of underlying continuous variables, we formulate a model which allows the comparisons of locations and dispersions between variables. We identify the model by imposing stochastic constraints on the thresholds that define the relationship between the observed and the underlying variables. As a result, the underlying continuous variables' location and dispersion parameters which were not estimable before can be estimated by the Bayesian approach. Illustrative examples are given based on several reported data sets. PMID- 10380318 TI - Measuring change: mixed Markov models for ordinal panel data. AB - In an analysis of longitudinal data it is important to distinguish between dependencies caused by within- and between-subject variability. This paper presents mixed Markov models for ordinal data that take into account both sources of variation. In addition, covariates that may capture differences among panel members and time-specific changes are also incorporated in the model. The model is derived by specifying an observation-driven process at the individual level and allowing for parametric or semi-parametric representations of random parameter variation across the units of analysis. As a result, the approach is well suited for modelling ordinal panel data with a large number of time points. In an application a three-week diary study is analysed to test hypotheses about the relationships between emotions and personality factors over time. PMID- 10380319 TI - [Suffering from lithiasis in the accounts of Michel de Montaigne and Cecilia Ferrazzi(based on investigations by Anne Jacobson Schutte)]. PMID- 10380320 TI - [Molecular forms of PSA and differentiating between benign hypertrophy,carcinoma, and disseminated prostatic carcinoma. Correlation between two determination technics]. AB - OBJECTIVE: To determine the correlation between two methods of measurement of total PSA (PSA-T) and free PSA (PSA-F) and the utility of the PSA-F/PSA-T ratio in patients with PSA-T between 4 and 10 ng/ml suspected as having prostate cancer. METHODS: Determinations of both PSA-T and PSA-F were performed using two different techniques in 91 patients suspected as having prostate cancer. Diagnosis was made on the findings of biopsy and the complementary tests. RESULTS: The following correlation was found for the two techniques: R = 0.99 and p < 0.05 for PSA-T, and R = 0.85 and p < 0.05 for PSA-F. For PSA-T values of 4-10 ng/ml and PSA-F greater than 25%, we found two patients with prostate cancer. For a PSA-F/PSA-T ratio less than 9%, all cases had prostate cancer. Three cases with bone metastasis had PSA-T values less than 10 ng/ml. CONCLUSIONS: A PSA-F/PSA-T ratio greater than 25% does not exclude malignancy in patients with a total PSA of 4-10 ng/ml; values less than 9% correspond to prostate cancer. Bone metastasis was found with both methods in patients with total PSA values less than 10 ng/ml. PMID- 10380321 TI - [Analysis of transfusion in kidney surgery]. AB - OBJECTIVE: To analyze the likelihood of perioperative transfusion in patients undergoing renal surgery. METHODS: The records of all patients who had undergone renal surgery in 1996 were reviewed. The abstracted patient discharge records, encoded according to the ICD-9-CM, were analyzed for gender, age, diagnosis, surgical and non-surgical procedures including transfusions of blood products. RESULTS: 21 (23.3%) of 90 patients who had undergone renal surgery required transfusion. By univariate analysis, the incidence of transfusion was statistically significantly higher in patients with chronic nephropathy, patients undergoing total nephrectomy and renal transplantation, and those having a greater number of ICD-9-CM diagnosis codes (indirect indicator of the clinical severity and comorbidity). By multivariate logistic regression analysis, the surgical procedure was found to be the only predictive factor for blood transfusion (p = 0.013). The main diagnosis, number of diagnosis codes, gender, number of surgical techniques and age were not found to be predictors of transfusion. CONCLUSIONS: Our data show that of the variables analyzed, the surgical procedure is the only predictive factor of the likelihood of requiring blood transfusion. PMID- 10380322 TI - [Intrascrotal lithiasis: an infrequent finding?]. AB - OBJECTIVE: To report on 10 additional cases of intrascrotal calculi and briefly review the literature and pathogenesis of this benign lesion. METHODS: 10 patients that had consulted for diverse testicular conditions were evaluated by ultrasound using the 7.5 MHz probe. RESULTS: All patients were found to have a hydrocele of a larger or smaller volume with a mobile hyperechoic focus that produced acoustic shadows. CONCLUSIONS: The ultrasound finding of intrascrotal calculi is becoming increasingly more frequent. In our view, this is due to the fact that more sonographic studies are currently performed. The possibility to diagnose this condition obviates the need for subsequent explorations or surgery. PMID- 10380323 TI - [Upper urinary tract tumors: results of treatment and follow-up]. AB - OBJECTIVE: To present the results of treatment and follow-up of 105 patients with tumor of the upper urinary tract. METHODS: A retrospective study was conducted on 105 patients (88 male and 17 female; mean age 68.3 years) with tumor of the upper urinary tract that had been treated from 1975 to 1997. In total 114 functional units were treated, including recurrences and bilateral tumors. The sites of involvement were: ureter (49.9%), pelvis (41.2%) and the entire upper urinary tract (8.7%). Ninety-six percent were transitional cell carcinomas: 4.8% were well differentiated (GI), 68% moderately differentiated (G2) and 26.8% poorly differentiated (G3); 58.6% were superficial, while 41.3% showed tumor invasion into or beyond the muscle layer. Ninety-two of the 105 patients were followed. The SPSS program was employed for the statistical analysis. The survival was calculated by the Kaplan-Meier method and the differences by the log rank test. Multivariance analysis was performed using the Cox regression method. RESULTS: TREATMENT: 58% underwent radical nephroureterectomy, 30% were treated conservatively and 11.6% underwent partial resection of the upper urinary tract. Recurrence: 8.7% of the patients showed tumor recurrence. The recurrence rate after conservative surgery was 13.6% and was as high as 80% in the remaining ureter. Metastasis: 22.8% of the patients presented metastasis to the retroperitoneal lymph nodes, bone, liver and lungs. Survival: In the univariate analysis tumor stage, age, radical and conservative surgery were found to influence survival, while stage and surgery (radical or conservative) were found to be statistically important by multivariate analysis. CONCLUSIONS: The treatment of choice for high grade and stage transitional cell carcinoma is by radical surgery, whereas for the superficial and well differentiated tumors, conservative management can achieve similar survival rates while preserving the renal unit and upper urinary tract. PMID- 10380324 TI - [Effect of prophylactic treatment with intravesical epirubicin in recurrence of superficial bladder tumor]. AB - OBJECTIVES: To determine the efficacy in the prevention of tumor recurrence, disease free interval and side effects of intravesical epirrubicin post-TUR of superficial bladder tumor. METHODS: From February 1993 to November 1997, 57 patients with stage Ta, T1, grade I, II and III tumor of the bladder were studied. These patients were treated with 50 mg epirrubicin in 50 ml saline solution administered intravesically on 15 occasions for one year after surgery. The mean follow-up was 18 months. The follow-up protocol included clinical, analytical, cytological, US and cystoscopic control evaluations every 3 months for the first year, every 6 months for the second and third year, and yearly thereafter. RESULTS: At a mean follow-up of 30.2 months, 31.5% (18/55) of the patients showed tumor recurrence; the disease free interval was 17 months for the remaining 18 patients and 43 months for the overall group. Cystitis was observed as an adverse effect in 5 of 57 patients (8.8%). CONCLUSIONS: The study shows that prophylactic treatment with intravesical epirrubicin for the prevention of recurrence in superficial bladder tumor has few side effects and its efficacy is similar to that reported in other studies. PMID- 10380325 TI - [Evaluation of new intraurethral occlusive depot (ORIS FEMININO) in the management of female stress urinary incontinence]. AB - OBJECTIVE: To determine the utility and safety of an intraurethral device (Oris Femenino) in the management of female stress urinary incontinence. METHODS: A prospective clinical study was conducted on 54 female patients, mean age 46.4 years, with stress urinary incontinence. The degree and severity of the incontinence was determined before and one month after treatment with the intraurethral device. The reasons for the dropouts were analyzed and the rehabilitative effects two months after treatment had been completed were evaluated in 22 patients. RESULTS: 83% of the patients who completed the study referred positive results. A significant improvement was demonstrated for the degree of incontinence as well as the number of absorbent pads used. Age and severity of incontinence inversely correlated with positive results. A correlation between the absence of a previous urethropexy and positive results was also found. The dropout rate was 44% (24 patients) and was chiefly due to symptomatic bacteriuria in 14 cases and the difficulty in fixation and degree of incontinence. A statistically significant reduction was found in the number of pads used in the group of patients evaluated two months after treatment had been completed. CONCLUSIONS: The intraurethral device analyzed in this study significantly reduced urinary incontinence. This method appears to be more effective in younger women who are not severely incontinent and have not previously undergone urethropexy. The intraurethral device appears to have some rehabilitative effect on the perineal muscle. Symptomatic bacteriuria was found to be the main disadvantage. PMID- 10380326 TI - [Treatment of ureteral obstruction with auto-expandable metallic endoprosthesis]. AB - OBJECTIVE: To evaluate the efficacy of Nitinol (Memotherm) self-expandible metal stents in the treatment of malignant and non-malignant ureteral obstruction in patients who are not eligible for surgery. METHODS: A total of 14 ureteral strictures (5 malignant, 9 non-malignant) in 13 patients were treated by the implantation of a Nitinol endoprosthesis. Eight were implanted by the antegrade and 6 by the retrograde route. A mean of 1.3 stents were implanted per patient. RESULTS: 85% of the stents were patent at a mean follow-up of 10.2 months (range 2-28). Four prostheses developed transient obstruction that was resolved by insertion of a double-J catheter for periods that ranged from 2 to 6 months. One stent required a permanent double-J catheter and another stent showed functional obstruction, although it was morphologically patent. CONCLUSION: The Nitinol self expandible metal stent is effective in the treatment of malignant and non malignant ureteral obstructions in patients who are not eligible for surgery due to the tumor stage or high surgical risk. PMID- 10380327 TI - [Voice-directed robotic cystoscopy]. AB - OBJECTIVE: In line with our previous studies in the field of robotics, a new application of our robotic arm is presented: voice-directed cystoscopy. METHODS: Cystoscopy was performed in a sow using a voice-directed robotic arm to which a cystoscope had been attached. The computer executes all the processes that enable interaction with the surgeon and communication with the robotic system. The surgeon directs the movements of the cystoscope by voice, using instructions easily recognizable by the voice identifier. RESULTS/CONCLUSIONS: This system is easy to operate and carries out the commands given by the surgeon with great precision and safety. PMID- 10380328 TI - [Hemorrhagic prostatic cyst following ultrasound guided biopsy. A case report]. AB - OBJECTIVE: To describe a case of a hemorrhagic prostatic cyst following ultrasound-guided biopsy of the prostate gland. METHODS/RESULTS: We reviewed our series of 77 patients submitted to re-biopsy of the prostate; only one case (1.3%) of hemorrhagic post-biopsy prostatic cyst was found. The ultrasound features, differential diagnosis and management of these cystic lesions are discussed. CONCLUSIONS: Hemorrhagic post-biopsy prostatic cyst is rare in our series (1.3%). A history of a previous biopsy in the area of the cystic lesion, the results of punction-aspiration of the cystic content and biopsy of the prostate gland provide data that are necessary to make the diagnosis. PMID- 10380329 TI - [Giant renal angiomyolipoma]. AB - OBJECTIVE: To present a cae of giant renal angiomyolipoma that required surgical treatment owing to its size and concomitant intratumoral hemorrhage. METHODS: The characteristics of the case are presented and discussed. RESULTS/CONCLUSIONS: Giant renal angiomyolipoma is one of the most frequent causes of Wunderlich syndrome and is diagnosed by ultrasound and CT. The decision to operate is based on the size of the lesion and/or symptomatology, as in the case described herein, which required a simple nephrectomy. PMID- 10380330 TI - [Inflammatory bladder pseudotumor]. AB - OBJECTIVE: To present a case of inflammatory pseudotumor of the urinary bladder, with special reference to the differential diagnosis. METHODS/RESULTS: A 26-year old patient with a history of gross hematuria and voiding syndrome underwent partial cystectomy for a bladder tumor. Pathological analysis disclosed an inflammatory pseudotumor of the bladder. The patient's postoperative course has been satisfactory. PMID- 10380331 TI - [Castleman's disease in the retroperitoneal region]. AB - OBJECTIVE: To describe a case of Castleman's disease presenting as a retroperitoneal mass, with special reference to the differential diagnosis from other retroperitoneal lesions. The histological features, variants, clinical manifestations, etiopathogenesis and treatment of Castleman's disease are reviewed. METHODS: A 64-year-old male presented with voiding symptoms and hypogastric pain. An ultrasound scan showed a 5 x 6 cm mass located behind the bladder and above the prostate, which was confirmed by an abdomino-pelvic CT scan. RESULTS: Retroperitoneal sarcoma was suspected and the mass was resected. The histopathological analysis showed giant lymphoid hyperplasia (vascular hyaline variant of Castleman's disease). CONCLUSIONS: Retroperitoneal Castleman's disease is a lymphoproliferative disorder with two well-defined histological types and a mixed variant. Although this lesion is frequently localized to the mediastinum (71%), extrathoracic lesions have been described. Definitive diagnosis is based on the postoperative pathological findings. PMID- 10380332 TI - [Intrascrotal malignant fibrous histiocytoma]. AB - OBJECTIVE: To describe a rare case of intrascrotal malignant fibrous histiocytoma. METHODS: A 90-year-old man presented with a scrotal mass which he had noted during the past few months. A scrotal tumor, 8 cm in diameter, was detected on physical evaluation. Tumor resection was decided due to the patient's good general condition despite his age. RESULTS: The histological analysis confirmed a sarcomatoid neoplasm with pleomorphism. The immunohistochemical analysis demonstrated strong overexpression of vimentin, alpha-1 antichymotrypsin, CD68 (Kpl), p53 oncoprotein and elevated Ki67. The ultrastructural study showed undifferentiated mesenchymal cells. The diagnosis of intrascrotal malignant fibrous histiocytoma was made based on the foregoing findings. CONCLUSIONS: Malignant fibrous histicytoma should be included in the differential diagnosis of intrascrotal tumors. Histological, immunohistochemical and ultrastructural studies are very important to distinguish the different paratesticular sarcomas. The strong overexpression of the p53 oncoprotein may be involved in the pathogenesis of this tumor type. PMID- 10380333 TI - [Bilateral hydronephrosis with papillary polypoid cystitis]. AB - OBJECTIVE: A case of papillary polypoid cystitis with bilateral hydronephrosis and an intravesical mass is presented. METHODS/RESULTS: A routine ultrasound evaluation performed in a 64-year-old diabetic female revealed an intravesical mass and bilateral hydronephrosis. There was extensive bladder involvement and the ureteral orifices could not be visualized during endoscopy. Transurethral resection of the lesion was performed. The histological examination showed papillary polypoid cystitis. The urinary cultures were positive, antibiotic therapy was instituted and the hydronephrosis, disappeared. CONCLUSIONS: Papillary polypoid cystitis, although a benign and superficial lesion, can be diffuse, involve both ureteral orifices and cause hydronephrosis. PMID- 10380334 TI - [Primary testicular tumor]. AB - OBJECTIVE: To report a case of primary germ cell testicular tumor that spontaneously remitted after metastasizing to the retroperitoneum, leaving histologically characteristic lesions of a fused tumor. The characteristic testicular ultrasound findings that permitted the diagnosis are described. METHODS/RESULTS: A 21-year-old male consulted for a retroperitoneal mass. The testes were normal on palpation. The testicular ultrasound showed a hyperechoic area with an acoustic shadow suspected as being a tumoral cicatrix. The histological analysis after orchidectomy revealed a mixed non-seminomatous germ cell tumor. CONCLUSIONS: Testicular ultrasound evaluation should be performed on all patients with retrosperitoneal germ cell tumor and normal testes on palpation. In the case described, there was good correlation between the ultrasound and the histological findings. PMID- 10380335 TI - AMS 800 artificial sphincter: an unusual case of circumscribed peritonitis due to prosthetic reservoir infection. AB - OBJECTIVES: Male urinary incontinence is nowadays a rare event in patients submitted to radical prostatectomy. In these cases, insertion on a hydraulic prosthesis is often the only therapeutical solution. Complications following this type of surgery are rare and when they occur, generally depend on the bacterial contamination of the device which will be "rejected". The cuff is generally the first cause of infection and its prompt removal should solve the problem in the majority of the cases. METHODS: Herein we report the case of a patient submitted to cuff removal three years earlier who consulted for a symptomless circumscribed peritonitis due to bacterial contamination of the intraperitoneal reservoir. RESULTS AND CONCLUSION: This report emphasizes the need to re-examine the indications for intraperitoneal implantation of the reservoir. Moreover, in case of a three-component prosthetic device, the external components (pump, cylinders and/or cuff) should be removed in a single step, at the same time, in order to avoid future contamination of the internal components. PMID- 10380336 TI - Improvement of side-chain modeling in proteins with the self-consistent mean field theory method based on an analysis of the factors influencing prediction. AB - With the objective of improving side-chain conformation prediction, we have analyzed the influence of various factors on prediction by the Self-Consistent Mean Field Theory method, applied to a set of high resolution x-ray protein structure models. These factors may be classed as variations in the mean field optimization protocol, variations in the potential energy function, and variations in rotamer library completeness. We have developed an optimization protocol that consistently reached lower mean field conformational free energies than two other protocols. This protocol led to an important improvement in prediction. We observed a major improvement in prediction with two more detailed van der Waals parameter sets, which we found to be due mainly to the introduction of scaling of 1-4 interactions. In a comparison of two knowledge-based rotamer libraries of considerably different size, we observed an unexpected decrease in prediction with an increase in library completeness. However, when we introduced a torsion potential term in the potential energy function, we found an important increase in average prediction and in the prediction of almost all residue types with a more complete rotamer set. The two knowledge-based rotamer libraries now became equivalent in terms of average prediction. The results we obtained in an analysis of the effect of the introduction of an additional electrostatic term in the potential energy function were largely inconclusive. However, we found a small increase in average prediction for an electrostatic potential term with a fixed dielectric constant of 15. The combined effect of all the factors we analyzed in this study resulted in average prediction accuracies of 79.9% for X1, 68.1% for X1 + 2, and 1.590 A for global rms deviation (RMSD); the corresponding values for core residues were 88.2%, 78.6%, and 1.171 A. These values represent improvements in average prediction of 6.5% for X1, 9.1% for X1 + 2, and 0.163 A for global RMSD over the original conditions; the corresponding improvements in the core were 5.9%, 9.0%, and 0.180 A, respectively. PMID- 10380337 TI - Hydration effects on the electrostatic potential around tuftsin. AB - The electrostatic potential and component dielectric constants from molecular dynamics (MD) trajectories of tuftsin, a tetrapeptide with the amino acid sequence Thr-Lys-Pro-Arg in water and in saline solution are presented. The results obtained from the analysis of the MD trajectories for the total electrostatic potential at points on a grid using the Ewald technique are compared with the solution to the Poisson-Boltzmann (PB) equation. The latter was solved using several sets of dielectric constant parameters. The effects of structural averaging on the PB results were also considered. Solute conformational mobility in simulations gives rise to an electrostatic potential map around the solute dominated by the solute monopole (or lowest order multipole). The detailed spatial variation of the electrostatic potential on the molecular surface brought about by the compounded effects of the distribution of water and ions close to the peptide, solvent mobility, and solute conformational mobility are not qualitatively reproducible from a reparametrization of the input solute and solvent dielectric constants to the PB equation for a single structure or for structurally averaged PB calculations. Nevertheless, by fitting the PB to the MD electrostatic potential surfaces with the dielectric constants as fitting parameters, we found that the values that give the best fit are the values calculated from the MD trajectories. Implications of using such field calculations on the design of tuftsin peptide analogues are discussed. PMID- 10380338 TI - Monte Carlo simulation of 4-alpha-glucanotransferase reaction. AB - 4-alpha-Glucanotransferase (GTase, D-enzyme) catalyzes disproportionation between two short polymers of maltooligosaccharides linked by alpha-1,4-glucoside bonds. Using action modes of the potato GTase for the donor and acceptor substrates, the Monte Carlo method was applied to simulate the GTase reaction. The simulation starts from a single enzyme molecule and a finite number (10(5)) of substrate molecules. All selection processes were performed using random numbers produced by computer. The initial substrates were from trimer to 10-mer. In every case, the final stage was the steady-state distribution of polymers. The steady-state distribution by the potato GTase reaction was different from those by the hypothetical random disproportionation reaction. The simulated data from the reaction of potato GTase and trimer almost quantitatively agreed with experimental data. The mechanism of the GTase reaction was accumulation of probabilistic processes and was well simulated by the Monte Carlo method. GTase randomizes the overall distribution of chain length of the substrate. Therefore the GTase reaction is an entropy-driven process. PMID- 10380339 TI - Light scattering, CD, and ligand binding studies of ferrihemoglobin polyelectrolyte complexes. AB - Quasi-elastic light scattering (QELS), electrophoretic light scattering (ELS), CD spectroscopy, and azide binding titrations were used to study the complexation at pH 6.8 between ferrihemoglobin and three polyelectrolytes that varied in charge density and sign. Both QELS and ELS show that the structure of the soluble complex formed between ferrihemoglobin and poly(diallyldimethylammonium chloride) [PDADMAC] varies with protein concentration. At fixed 1.0 mg/mL polyelectrolyte concentration, protein addition increases complex size and decreases complex mobility in a tightly correlated manner. At 1.0 mg/mL of greater protein concentration, a stable complex is formed between one polyelectrolyte chain and many protein molecules (i.e., an intrapolymer complex) with apparent diameter approximately 2.5 times that of the protein-free polyelectrolyte. Under conditions of excess polyelectrolyte, each of the three ferrihemoglobin polyelectrolyte solutions exhibits a single diffusion mode in QELS, which indicates that all protein molecules are complexed. CD spectra suggest little or no structural disruption of ferrihemoglobin upon complexation. Azide binding to the ferrihemoglobin-poly(2-acrylamide-2-methylpropanesulfonate) [PAMPS] complex is substantially altered relative to the polyelectrolyte-free protein, but minimal change in induced by complexation with an AMPS-based copolymer of reduced linear charge density. The change in azide binding induced by PDADMAC is intermediate between that of PAMPS and its copolymer. PMID- 10380340 TI - New results concerning the behavior of cellulose acetate in solutions and films by means of CD measurements. AB - Cellulose acetate (DS = 2.45) was extensively investigated by Circular Dichroism (CD) in acetonitrile and dioxane. We found great differences between the CD spectra of a 1 wt % acetonitrile solution and the corresponding dioxane solution of cellulose acetate (CA) indicating that the macromolecules exist in those solutions in different molecular arrangements (e.g., persistence length, solvatation shell). The resulting morphologies could be transformed reversibly into each other, as we found by measuring the CA in mixtures of both solvents. Solid CA films show discernible CD spectra depending on the solvent they were evaporated from. In this way, we prepared solid films of the same polysaccharide owning different chiral properties. Furthermore, changes in the spectra occurred with increasing CA concentration. Basing upon our findings, some general statements concerning the polymer behavior of CA are possible. PMID- 10380341 TI - Basic conformers in beta-peptides. AB - The conformation of oligomers of beta-amino acids of the general type Ac-[beta Xaa]n-NHMe (beta-Xaa = beta-Ala, beta-Aib, and beta-Abu; n = 1-4) was systematically examined at different levels of ab initio molecular orbital theory (HF/6-31G*, HF/3-21G). The solvent influence was considered employing two quantum mechanical self-consistent reaction field models. The results show a wide variety of possibilities for the formation of characteristic elements of secondary structure in beta-peptides. Most of them can be derived from the monomer units of blocked beta-peptides with n = 1. The stability and geometries of the beta peptide structures are considerably influenced by the side-chain positions, by the configurations at the C alpha- and C beta-atoms of the beta-amino acid constituents, and especially by environmental effects. Structure peculiarities of beta-peptides, in particular those of various helix alternatives, are discussed in relation to typical elements of secondary structure in alpha-peptides. PMID- 10380342 TI - An NMR and conformational investigation of the trans-syn cyclobutane photodimers of dUpdT. AB - Both trans-syn cyclobutane-type photodimers of 2'-deoxyuridylyl (3'-5') thymidine (dUpdT) were formed by deamination of the corresponding trans-syn cyclobutane photodimers of 2'-deoxycytidylyl (3'-5') thymidine (dCpdT) and were examined by 1H-, 13C-, and 31P-nmr spectroscopy. One- and two-dimensional nmr experiments provided a nearly complete assignment of the 1H, 13C, and 31P resonances. Scalar and nuclear Overhauser effect contacts were used to determine the conformation of the deoxyribose rings, exocyclic bonds, cyclobutane rings, and glycosidic linkages. Isomer I (S-type class; CB-; SYN-ANTI) and isomer II (N-type class; CB+; ANTI-SYN) exhibit markedly different conformational features. 31P chemical shifts show that the relative flexibility is dUpdT > isomer II > isomer I. The conformations of these species are very similar to those of other previously examined trans-syn photodimers. Among bipyrimidine photodimers of a given diastereomeric form (i.e., trans-syn I or II), the nmr-derived conformational parameters are nearly invariant, regardless of base substitution pattern. This contrasts with the substituent-dependent variation of cyclobutane ring conformation observed by Kim et al. (Biopolymers, 1993, Vol. 33, pp. 713-721) for an analogous series of cis-syn photodimers. Steric crowding of cyclobutane ring substituents is offered as an explanation for the difference in substituent effects between the families of cis-syn and trans-syn photodimers. PMID- 10380343 TI - Enzymatic ligation for synthesis of single-chain analogue of monellin by transglutaminase. AB - Monellin, a sweet protein, consists of two noncovalently associated polypeptide chains: an A chain of 44 amino acid residues and a B chain of 50 residues. Microbial transglutaminase (MTGase) was used for ligation of the monellin subunits without any protecting groups, and without activation of the C alpha carboxyl group at the C-terminus. Since a peptide fragment LLQG is a good substrate for MTGase to form an amide bond between the gamma-amide group of the Gln residue and the epsilon-amino group of Lys, a monellin B chain analogue in which LLQG was elongated at the C-terminus (B-LLQG) was synthesized by solid phase synthesis. The monellin A chain analogue in which KGK was elongated at the N-terminus (KGK-A) was synthesized by the same method as that of the B chain analogue. The KGK-A chain and the B-LLQG chain were coupled by MTGase to give single-chain analogue of monellin. The single-chain analogue of monellin was characterized by analytical reverse phase high performance liquid chromatography, electrospray ionization, and amino acid analyses. All analyses gave satisfactory results. The single-chain analogue of monellin was more heat stable than natural monellin. PMID- 10380344 TI - Study of the interactions of D- and L-polylysine enantiomers withpectate in aqueous solutions. AB - The interaction between D- and L-enantiomers of polylysine and potassium pectate was studied by means of CD, microcalorimetry, and osmometry. Upon binding with pectate, only poly(L-lysine) undergoes a coil to alpha-helix transition, while poly(D-lysine) remains in the disordered state. This suggest that the energetics of the interaction is influenced by stereochemical constraints besides electrostatic forces. Experimental findings from microcalorimetry suggest that a contribution to the overall enthalpy of binding comes from the polysaccharidic moiety. Stoichiometry of the macromolecular complexes studied by osmometry gives a polylysine:pectate ratio of 3:1, in agreement with the respective degree of polymerization of the two polyelectrolytes. PMID- 10380345 TI - Cyclic pentapeptides of chiral sequence DLDDL as scaffold for antagonism of G protein coupled receptors: synthesis, activity and conformational analysis by NMR and molecular dynamics of ITF 1565 a substance P inhibitor. AB - Under the hypotheses of a structurally related binding site for antagonists of G protein coupled receptors and the ability of cyclic pentapeptides of chiral sequence D1L2D3D4L5 to form rigid structures with which probe the pharmacophoric specificity of these receptors, inhibitors of substance P were designed based on available structure-activity relationships. ITF 1565, cyclo[D-Trp1-Pro2-D-Lys3-D Trp4-Phe5], antagonized substance P activity mediated by type 1 neurokinin receptor (NK1) whereas it acted weakly against NK2 and did not inhibit endothelin at all. The preferential conformation of the peptide was obtained from nmr spectroscopy and computer calculations, and shown to contain the same beta II turn and gamma'-turn found in other cyclic pentapeptides with the same chiral sequence. The structure of the peptide was compared with that of the beta-D glucose molecule that has been proposed as a semirigid scaffold for antagonists of G-protein coupled receptors. The gamma'-turn of the cyclic peptide superimposed well with beta-D-glucose in the chair conformation. Furthermore, when the side chains were considered, the aromatic groups of the two molecules were found to generally overlap. These results support the view of G-protein coupled receptors as possessing structurally similar binding sites for antagonists and suggest that cyclic pentapeptides of chiral sequence D1L2D3D4L5 may be useful as semirigid scaffolds for the design of antagonists of this family of receptors. PMID- 10380346 TI - Enzymatic generation of binary block-copolymeric structures: mathematical analysis based on triad frequencies evaluated by NMR. AB - A mathematical model is derived for describing a multiple-attack pathway for enzymatic generation of block structure in binary linear copolymers having initially a randomized sequential structure. The model is based on sequential information in terms of copolymer monads, diads, and triads estimated by nmr spectroscopy, and is applicable to enzymes attacking next to a reacted unit in the polymer chains. Then the block distribution of unreacted units remains constant and explicit relationships are provided. The probability of triad frequencies as a function of monads, i.e., progress curve of enzyme copolymer sequential structure, allows us to characterize the enzymatic mode of attack independently of enzyme kinetics. The produced fractions of heterogeneous triads centered by reacted units are shown to be affected, to a large extent, by the degree of multiple attack (d) entering into the formula as a variable parameter. The single-chain, d = infinity, and multiple-chain mechanisms, d = 1, representing the two extremes of the treated mechanism, are very clearly discriminated. PMID- 10380347 TI - Protease inhibitors. PMID- 10380348 TI - Design of protease inhibitors on the basis of substrate stereospecificity. AB - The substrate stereospecificity in enzymic reactions, which is one of characteristics of enzymes along with the substrate and regiospecificity can provide a basis for the rational design of inhibitors. This has been demonstrated using alpha-chymotrypsin, a prototypic serine protease as a model enzyme. On the basis of the structure-activity relationships for substrates as well as inhibitors and mechanism of the enzymic reaction, a schematic three-dimensional model of the S1 subsite of alpha-chymotrypsin is constructed. It was envisioned from the three-dimensional active site model that 2-benzyl-3,4-epoxybutanoic acid methyl ester (1) having a (2S)-configuration would bind the enzyme with its oxirane ring being rested at the catalytic site, in which the oxirane ring is subject to a nucleophilic attack by the Ser-195 hydroxyl to form a ether linkage. Kinetic analysis of the enzymic reaction in the presence of the potential inhibitors showed that (2S,3R)-1 inactivates alpha-chymotrypsin, while (2S,3S)-1 inhibits the enzyme competitively. The lack of inactivating activity in the case of (2S,3S)-1 may be due to the unfavorable alignment of the C3-O bond with respect to the hydroxyl of Ser-195 for the SN2-type ring cleave reaction of the oxirane moiety. When the design protocol was applied to papain, a representative cysteine protease, (2S,3S)-1 inhibited the enzyme irreversibly, while (2S,3R)-1 inhibited reversibly. On the basis of the stereospecificity shown in the inactivation of the enzymes, it was inferred that in the case of alpha chymotrypsin, the nucleophilic attack of the Ser-195 hydroxyl at the scissile carbonyl carbon of substrates occurs in a si fashion, while the thiolate of Cys 25 in papain attacks the substrate amide bond in a re fashion. The inhibitor design protocol may be applied to other proteases. PMID- 10380349 TI - Design of serine protease inhibitors with conformation restricted by amino acid side-chain-side-chain CH/pie interaction. AB - A novel type of conformationally restricted peptides with the structure of H-D Xaa-Phe-NH-CH2-C6H5 has been developed as inhibitors of serine proteinase chymotrypsin. The D-Xaa-alkyl and Phe-phenyl groups resulted in a formation of the hydrophobic core due to the side-chain-side-chain CH/pie interaction. Their spatial proximity was evidenced by 400 MHz 1H-nmr measurements, observing large upfield shifts of proton signals of D-Xaa-alkyl and nuclear Over-hauser effect (NOE) enhancements between the D-Xaa-alkyl and Phe-phenyl groups. This conformational restriction brought by CH/pie interaction produced an inhibitory structure, in which the C-terminal amide-benzyl group fits the chymotrypsin S1 site and the hydrophobic core binds to the S2 site. The inhibitory conformation was demonstrated crystallographically for the complex between the dipeptide H-D Leu-Phe-NH-CH2-C6H4(p-F) and gamma-chymotrypsin. Detailed structure-activity studies have substantiated the structure of dipeptides in the active center of the enzyme. PMID- 10380350 TI - From natural to synthetic multisite thrombin inhibitors. AB - A large number of potent and selective therapeutic agents, useful for the treatment of several diseases, have been isolated from natural sources. For example, the most active thrombin inhibitors are those secreted by the salivary glands of leeches. One peculiar feature of these agents is the lack of any significant inhibitory cross-reaction with other serine proteinases. Hence, the knowledge of the exact mechanism of action of these molecules provides the basis for the development of new and efficient synthetic drugs. For this reason, many studies have been undertaken on the structure-activity relationships of natural thrombin inhibitors, and a large amount of detailed information has been obtained by the crystal structures of these inhibitors when complexed with thrombin. In this paper, we review natural and synthetic multisite thrombin inhibitors, whose structural aspects have been determined in detail. We also report here the approach used by us to develop a new class of synthetic, multisite directed thrombin inhibitors, named hirunorms, designed to mimic the distinctive binding mode of hirudin. PMID- 10380351 TI - Development of plasma kallikrein selective inhibitors. AB - During the course of the development of active center-directed plasmin inhibitors, it was found that N-(trans-4-aminomethylcyclohexanecarbonyl)-lysine-4 methoxycarb onylanilide inhibited plasma kallikrein more potently than other enzymes such as plasmin, urokinase, and thrombin, although the inhibitory activity was not as potent and enzyme selectivity not as high. Based on studies of structure-activity relationship, we designed and synthesized the plasma kallikrein selective inhibitor, N-(trans-4-aminomethylcyclohexanecarbonyl) phenylalanine-4-carboxy methyl- anilide (Tra-Phe-APAA). Tra-Phe-APAA inhibited plasma kallikrein with a Ki value of 0.81 microM, while it inhibited glandular kallikrein, plasmin, urokinase, tissue plasminogen activator, factor Xa, factor XIIa, and thrombin with Ki values of > 500, 390, 200, > 500, > 500 > 500, and > 500 microM, respectively. We designated Tra-Phe-APAA as PKSI-527. Using PKSI-527 as an affinity ligand, we synthesized a new affinity gel (PKSI-Toyopearl) and employed it for the rapid purification of plasma kallikrein from human plasma. Human plasma activated with kaolin after acid treatment was applied to a PKSI-527 Toyopearl column. Adsorbed protein was eluted with 50 mM glycinehydrochloric acid buffer (pH 3.0). Plasma kallikrein was purified 181-fold with a yield of 85% from the kaolin-activated plasma. PMID- 10380352 TI - Structure-based discovery of Tipranavir disodium (PNU-140690E): a potent, orally bioavailable, nonpeptidic HIV protease inhibitor. AB - Efforts to develop therapeutically relevant HIV protease inhibitors as medicinal agents in confronting the AIDS crisis have been aided by the wealth of fundamental information acquired during related drug discovery campaigns against other aspartyl proteases. This knowledge base was brought to full force with the broad screening identification of small, nonpeptidic, inhibitory molecules as templates for chemical elaboration. Significantly, the ability to collect crystallographic data on the inhibitor-enzyme complexes in a rapid fashion afforded the opportunity for a structure-based approach to drug discovery. Iterative cycles of synthesis, biological testing, and structural information gathering followed by prudent design modifications afforded compounds suitable for clinical evaluation. Displaying high enzymatic inhibition (Ki = 8 pM), potent in vitro antiviral cell culture activity (IC90 = 100 nM), and a useful pharmacokinetic profile, PNU-140690E (Tipranavir disodium) has entered into clinical studies. Promising results from these early trials supported further evaluation of this compound in HIV-infected individuals. PNU-140690E is currently under extensive clinical study. PMID- 10380353 TI - Small dipeptide-based HIV protease inhibitors containing the hydroxymethylcarbonyl isostere as an ideal transition-state mimic. AB - The human immunodeficiency virus (HIV) codes for an aspartic protease known to be essential for retroviral maturation and replication. HIV protease is formed from two identical 99 amino acid peptides. We synthesized [(NHCH2CH2-S-CH2CO)51-52, Ala67,95]HIV-1 protease using the thioether chemical ligation method, and then prepared the [(NHCH2CH2-S-CH2CO)51-52, Ala67,95, Cys98]HIV-1 protease dimer analogue covalently linked by a disulfide bridge. These HIV-1 protease analogues effectively cleaved the Tyr-Phe-type substrate, but had weak affinity to the Tyr Pro-type substrate. Consequently, the molecular recognition of the protease analogues differs from that of the wild-type enzyme. Based on the substrate transition state, we designed and synthesized a novel class of HIV protease inhibitors containing an unnatural amino acid, (2S, 3S)-3-amino-2-hydroxy-4 phenylbutyric acid, named allophenylnorstatine, with a hydroxymethylcarbonyl (HMC) isostere. The stereochemistry of the hydroxyl group was significant for the enzyme inhibition and the HMC group interacted excellently with the aspartic acid carboxyl groups of HIV protease active site in the essentially same hydrogen bonding mode as the transition state. Small dipeptide-based HIV protease inhibitors containing the HMC isostere were studied as advantageous compounds. Among them, a dipeptide-based HIV protease inhibitor, KNI-577, exhibited potent antiviral activities, low cytotoxicity, and good pharmacokinetic properties. PMID- 10380354 TI - Comparison of inhibitor binding to feline and human immunodeficiency virus proteases: structure-based drug design and the resistance problem. AB - The design and synthesis of compounds targeted against human immunodeficiency virus 1 (HIV-1) protease have resulted in effective antiviral therapies. However, the rapid replication of the virus and the inherent mutability of the viral genome result in the outgrowth of resistant strains in the majority of patients. Thus, there is a continuing need to develop new antiprotease compounds that may bind more effectively to the resistant forms of protease. This contribution examines the binding of a single inhibitor to two different retroviral proteases, HIV-1 protease and feline immunodeficiency virus protease. Despite the overall similarity of the related retroviral enzymes, specific substitutions within the binding site cavity provide a distinctly different binding landscape that dramatically alters the affinity of compounds. Through this comparison, insights have been obtained into new strategies for drug design. New compounds based on these concepts have been tested against the two enzymes. PMID- 10380355 TI - The human mucus protease inhibitor and its mutants are novel defensive compounds against infection with influenza A and Sendai viruses. AB - Tryptase Clara, a trypsin-like protease localized exclusively in and secreted by Clara cells of the bronchial epithelium, is a prime host factor that processes viral envelope glycoproteins and determines the infectivity of influenza A and Sendai viruses (H. Kido, Y. Yokogoshi, K. Sakai, M. Tashiro, Y. Kishino, A. Fukutomi, and N. Katunuma, The Journal of Biological Chemistry, 1992, Vol. 267, pp. 13573-13579). We report here that human mucus protease inhibitor (MPI), a major inhibitor of granulocyte elastase in the lining fluid of the human respiratory tract, significantly inhibited induction of the infectivity of influenza A and Sendai viruses by tryptase Clara in vitro and multicycles of mouse-adapted influenza A virus replication in rat lungs in vivo. Recombinant MPI and the C- but not the N-terminal domain of MPI inhibited both the activity of tryptase Clara and the induction of virus infection by tryptase Clara. The 50% inhibitory concentrations of MPI and the C-terminal domain peptide (Pro50-Ala107) of MPI for tryptase Clara were 7.4 and 61.6 nM, respectively, with Sendai virus envelope glycoproteins as the substrate. Studies on deletion mutants of the C terminal domain of MPI revealed that the minimal size of MPI required for the inhibition of tryptase Clara is the peptide Lys60-Ala107. Studies involving site directed mutagenesis of the C-terminal domain of MPI indicated that the Leu72 Met73 site of MPI is the inhibitory site for tryptase Clara. Substitution of residue Leu72 with a basic amino acid significantly increased in the inhibitory activity of the C-terminal domain of MPI, but further substitution of residue Met73 with various amino acids in these mutants reduced the inhibitory activity. Since there is evidence suggesting that the concentration of MPI in respiratory fluid is insufficient for prevention of virus infection, the administration of MPI, the recombinant C-terminal domain of MPI, and their mutants, with residue Leu72 substituted with residues Arg72 and Lys72, may be useful for treatment of such pneumotropic virus infections. PMID- 10380356 TI - Inhibition of cysteine proteases by peptides containing aziridine-2,3 dicarboxylic acid building blocks. AB - Mammalian cysteine proteases of the papain superfamily are interesting targets for the development of new drugs against diseases connected to abnormal degradation of muscle or bone proteins. The high nucleophilicity of the active site of these proteases as well as the characteristics of the well-known epoxysuccinic acid derived cysteine protease inhibitors provided a basis for the design of new types of selective and irreversible inhibitors for these enzymes. We designed and synthesized a novel class of peptidic cysteine protease inhibitors containing aziridine-2,3-dicarboxylic acid as electrophilic amino acid. Three types of aziridinyl peptides that differ in the position of the aziridine building block within the peptide chain have been synthesized and tested as inhibitors of several cysteine proteases. Remarkable differences could be observed between the three types of inhibitors concerning their activity, stereospecificity, pH dependency of inhibition, and selectivity between different cysteine proteases, respectively, indicating that different binding modes of the three types of inhibitors in respect to their orientation in the S and S' binding sites of the enzymes may be present. PMID- 10380357 TI - Structural basis of inhibition of cysteine proteases by E-64 and its derivatives. AB - This paper focuses on the inhibitory mechanism of E-64 and its derivatives (epoxysuccinyl-based inhibitors) with some cysteine proteases, based on the binding modes observed in the x-ray crystal structures of their enzyme-inhibitor complexes. E-64 is a potent irreversible inhibitor against general cysteine proteases, and its binding modes with papain, actinidin, cathepsin L, and cathepsin K have been reviewed at the atomic level. E-64 interacts with the Sn subsites of cysteine proteases. Although the Sn-Pn (n = 1-3) interactions of the inhibitor with the main chains of the active site residues are similar in respective complexes, the significant difference is observed in the side-chain interactions of S2-P2 and S3-P3 pairs because of different residues constituting the respective subsites. E-64-c and CA074 are representative derivatives developed from E-64 as a clinical usable and a cathepsin B-specific inhibitors, respectively. In contrast with similar binding/inhibitory modes of E-64-c and E 64 for cysteine proteases, the inhibitory mechanism of cathepsin B-specific CA074 results from the binding to the Sn' subsite. PMID- 10380359 TI - [The care of acute pancreatitis: a still current challenge between medicine and surgery]. PMID- 10380358 TI - Peptidyl beta-homo-aspartals (3-amino-4-carboxybutyraldehydes): new specific inhibitors of caspases. AB - Interleukin-1 beta (IL-1 beta)-converting enzyme (ICE, caspase-1) processes the IL-1 beta precursor to mature inflammatory cytokine IL-1 beta. ICE has been identified as a unique cysteine protease, which cleaves Asp-X bonds, shows resistance to E-64 (an inhibitor of most cysteine proteases) and has a primary structure that is homologous to CED-3, a protein required for apoptosis (programmed cell death) in the nematode Caenorhabditis elegans, and to mammalian cysteine proteases that initiate and execute apoptosis, e.g., apopain/CPP32/caspase-3. The inhibitors of the ICE/CED-3 family or caspases, as they are called recently, may constitute therapeutic agents for amelioration of inflammatory and apoptosis-associated diseases. The most efficient ICE inhibitors are peptide aldehydes and peptidyl chloro or (acyloxy)methanes. A recent study revealed that both D- and L-Asp are accepted by ICE at the P1 of such inhibitors, and the peptidyl (acyloxy)methane analogues having the beta-homo-aspartyl residue [-NH-CH(CH2COOH)-CH2CO-] are inactive. These findings we reexamined in terms of two issues. (a) ICE's resistance to E-64. Since it was thought to be caused by the enzyme's unique substrate specificity, we prepared substrate-based analogues, which were not inhibitory suggesting significant structural difference between the active centers of ICE and papain-like enzymes. (b) Tolerance for D stereochemistry at the P1 of these inhibitors. In view of the mechanism of cysteine protease inhibition by peptidyl X-methanes, we thought that this phenomenon should be a general characteristic of cysteine proteases and the hAsp containing analogues should behave as reversible inhibitors. Here, we analyzed the inhibition of ICE and apopain in comparison with that of papain, thrombin, and trypsin by peptide L/D-alpha-aldehydes and their L-beta-homo-aldehyde [-NH CH(R)-CH2-CHO] analogues. The following results were found. (1) The peptidyl L beta-homo-aspartals are potent inhibitors for caspases. (2) The L-beta-homo analogues of peptide aldehyde inhibitors designed for other proteases are not inhibitory. (3) Unlike trypsin and thrombin (serine proteases), papain (cysteine protease) shows tolerance for D-stereochemistry at the P1 site of peptide aldehydes in proportion to the lability of the alpha-hydrogen of the P1-D residue. The complete tolerance of ICE for P1-D-Asp may arise from this residue's high tendency to epimerization. (4) Reaction of cysteine proteases with peptide aldehyde or peptidyl X-methane inhibitors containing P1-D-residues may include alpha-proton abstraction followed by asymmetric induction leading to P1-L-residue containing products. PMID- 10380360 TI - The Chievitz juxtaparotid organ. AB - The Chievitz juxtaparotid organ represents a macroscopic longitudinal formation, which is developed from oral cavity ectoderm in its lateral wall. As to its function, the organ probably represents a mechanosensor with different qualities of perception. The information coming from its sensors takes part in different activities of the lateral wall of oral cavity during sucking, swallowing, mastication, speech, protecting reflexes and wall tonus. The Chievitz juxtaparotid organ is not only a morphologically interesting structure, but is of great importance also for clinic and surgical pathology of the oral cavity. PMID- 10380361 TI - [The spontaneous rupture of a pyogenic liver abscess. A clinical case]. AB - The authors report a case of a large monolocular liver abscess in a patient with gallbladder and choledocic stones and biliary stent, complicated by rupture into the peritoneal cavity, that required surgical treatment. Surgical exploration versus other approach results the choice treatment, as shown by resolution of the pathology. PMID- 10380362 TI - [Isolated mesenteric fibromatosis. A clinical case]. AB - The authors report a case of isolated mesenteric fibromatosis un associated with familial adenomatous polyposis or Gardner's syndrome or prior abdominal surgery. These neoplasms are usually asymptomatic until when the compression of the small or large bowel or the ureter causes symptoms; although they are benign lesions without metastases, local recurrences are very frequent. Surgical removal is the primary treatment; until now no satisfactory results have been obtained with other therapeutic modalities. PMID- 10380363 TI - [Renal oncocytoma: its differential diagnosis and the indications for surgical treatment]. AB - The authors report an uncommon renal oncocytoma rate stressing difficulties that modern diagnostic modalities meet in a correct preoperative differential diagnosis with nephrocarcinoma. Oncocytoma is a low multifocal involvement. Surgical treatment is the primary choice, nephrectomy about localized monofocal lesions in advisable, with short and long term positive outcomes. In ambilateral involvement case without malignancy evident signs (scarce likelihood to infiltrate nephritic capsule, without lymphoadenopathy) conservative operations with partial resections are suggested with a five years survival from 84 to 96%. PMID- 10380364 TI - [Carotid body tumors. Apropos a case and a review of the literature]. AB - Chemodectomas are rare tumors arising from paraganglionic cells located at the level of carotid bifurcation. They are usually benign and non functioning, presenting as a slow growing cervical mass. A preoperative diagnosis is mandatory, based on doppler color flow imaging and angiography. Surgery is the only therapy providing total eradication of this tumor. Subadventitial resection is the most established technique, although resection of a large mass may require carotid replacement by interposition graft. Cranial nerve palsy and stroke are the perioperative complications most frequently encountered. The Authors report here a case of carotid body tumor and a review of the literature in order to define clinical characteristics of the tumor and proper diagnostic and therapeutic approaches to this rare neoplasm. PMID- 10380365 TI - [Tumefaction of the parotid region. The clinico-surgical experience of 3 years]. AB - The pathologic processes involving the parotid gland area include a vast, heterogeneous group of lesions, consisting of dysembryopathies, traumas, acute and chronic inflammation, degenerative manifestations, benign tumours and both primary and secondary malignancies. This gland, or rather the whole parotid gland area, can be a site of secondary invasion, due to the presence of intra- and peri parotid lymph nodes; the metastases usually deriving from small, sometimes unrevealed tumours. Treatment of parotid gland tumours is mainly surgical; in most cases the choice of therapy depends on the clinical features and the results of preoperative diagnostic tests. This paper presents our experience during the last three years of clinico-surgical activity in this field and discusses the treatment of primary and secondary tumours of the parotid gland area. PMID- 10380366 TI - [The complications of implantable cardioverter-defibrillators and their treatment]. AB - Infection of implantable cardioverter defibrillator (ICD) is a devastating event. In an effort to more fully understand ICD infection, the authors reviewed patients records recommending a strategy for management based on their satisfactory experience. From March 1993 through May 1998, 85 ICD were implanted in 64 male and 21 female patients. Transmediastinal approach was performed in 8 (9.5%) cases and transvenous in 77 (91.5%). All device-related infections were examined. Seven (8.25%) device-related infections occurred with a mean time interval of 3 months. In all cases bacterial infection was demonstrated. All infections involved the generator with or without other components involvement. First approach was conservative in all cases but it wasn't successful. Then the authors always used a surgical therapy, in 3 cases removing electrodes by traction and in 4 resorting to cardiopulmonary bypass (CPB). Two deaths were registered. Explantation of ICD resolved in all cases infective complications with no early or additional reinfections. In the last cases with devices implanted by transvenous approach and subpectoral generator implant, no infective complications were observed. In authors experience a complete removal of the ICD generator as well as of all its components is to be preferred as soon as the infections develops. PMID- 10380367 TI - [Surgery associated with intraoperative hyperthermia-chemotherapy in the treatment of malignant tumors of the digestive system with peritoneal carcinosis]. AB - In the last two years the authors have treated 4 patients affected with malignant tumors of large bowel with very poor prognosis. In every cases they found large spreading to peritoneal cavity. These patients underwent to intraperitoneal hyperthermia-chemotherapy using a special device conceived by authors. This treatment produced promising results as related to life quality in these cases. PMID- 10380368 TI - [The intracystic concentration of MCA in the fluid from large breast cysts]. AB - 75 patients with breast gross cystic disease and no cancer have been included in the study. For each patients serous and intracystic concentrations of MCA have been measured. The aim of the study is to assess whether if a relation between intracystic concentration of the marker and resistance and capability of cellular reproduction exists (confirmed by the release of the cyst). The analysis of intracystic values shows that synthesis of MCA is an intrinsic peculiarity of cytologic kind. It is apparently independent from inflammatory or hemorrhagic contemporary processes. PMID- 10380369 TI - Testing a biochemical model of human genetic resistance to falciparum malaria by the analysis of variation at protein and microsatellite loci. AB - We recently proposed a biochemical model of genetic resistance to falciparum malaria based on the role of oxidant stress (of parasitic origin) in inducing the irreversible oxidation of hemoglobin and its binding to the erythrocyte membrane (Destro-Bisol et al. 1996). To test the model, we analyzed the relationships between the polymorphisms at the hemoglobin beta chain (HBB) and red cell glutathione peroxidase (GPX1) loci in 18 populations that had been subjected to endemic malaria (Cameroon and Central African Republic). The erythrocytes of GPX1*2 heterozygotes should be more efficient in sheltering the cell membrane from irreversible oxidation and binding of hemoglobin caused by the oxidant stress exerted by Plasmodium falciparum. According to our model, the GPX1*2 allele has an epistatic effect on the HBB*A/*S genotype by lowering its protection against falciparum malaria. In turn, this should decrease the fitness of the HBB*A/*S-GPX1*2/*1 genotype. Our predictions were confirmed. In fact, we observed a clear trend toward a dissociation between the HBB*A/*S and GPX1*2/*1 genotypes in the overall data. To test alternative hypotheses, we also analyzed the genetic variation at 9 protein and 10 autosomal microsatellite loci at both the single- and the 2-locus level. We also discuss the possible relevance of an alternative biochemical pathway. The results further support the conclusions of our study because the dissociation between the GPX1*2/*1 and HBB*A/*S genotypes does not appear to be related either to a general decrease in heterozygosity or to an increased risk of sudden death in HBB*A/*S individuals. PMID- 10380370 TI - Some atypical and rare sickle cell gene haplotypes in populations of Andhra Pradesh, India. AB - We have investigated the clinical, hematological, and molecular genetic characteristics of sickle cell anemia patients from 6 populations of Andhra Pradesh, South India. Of 72 sickle cell chromosomes (HBB*S) 60 belong to characteristic Arab-Indian haplotypes, 6 to variant Arab-Indian haplotypes, 1 to a Bantu haplotype, 2 to a Cameroon haplotype, and 3 to rare haplotypes. This is the first report of a Bantu haplotype in an Indian population. Some information on haplotype characteristics of normal chromosomes (HBB*A) is also presented. The average hemoglobin level was 7.3 g% and mean fetal hemoglobin (HbF) level was 12.6%. The higher HbF levels corroborate earlier observations in sickle cell homozygotes from India. Clinical investigations have revealed splenomegaly and painful crises as the most common features in these patients. PMID- 10380371 TI - STR polymorphisms in the population of the island of Hvar. AB - The aim of this study is to analyze short tandem repeat (STR) variation using data on 9 loci (D3S1358, VWA, FGA, THO1, TPOX, CSF1PO, D5S818, D13S317, D7S820) from the subpopulations of 6 villages on the island of Hvar, Croatia. The STR data help us to analyze the genetic structure of Hvar. The analysis of STR data in this study indicated genetic homogeneity among the village subpopulations on Hvar and the lack of the so-called east-west dichotomy, which had been indicated by some previous multidisciplinary anthropological studies. The observed value of GST (0.030) is most probably a consequence of high STR mutation rates, which produce a high level of within-group (village) diversity relative to total diversity of the population. The validity of STR markers in assessing genetic structure of small populations and especially in determining the relationships among closely related and reproductively isolated groups remains to be further evaluated. PMID- 10380372 TI - HLA genes and haplotypes in Ryukyuans suggest recent gene flow to the Okinawa Islands. AB - Polymorphism of HLA genes was investigated in a population sample of Ryukyuans living on the main island of Okinawa (n = 197), in the southwestern islands of Japan. Serological typing was applied to class I loci (HLA-A, -B, and -C) and to HLA-DRB1; nucleotide sequence-level typing was performed using PCR microtiter plate hybridization and PCR single-strand conformation polymorphism methods. Ryukyuans showed a higher frequency of DRB1*0405 and lower frequencies of DRB1*1502 and DRB1*1302 compared with Hondo Japanese living on main islands. Principal components and phylogenetic analyses of 12 East Asian populations, including Ryukyuans, were performed based on the allele frequencies of HLA-A, -B, and -DRB1. In the principal components analysis 3 Japanese populations (Ryukyuans, Hondo Japanese, and Ainu) formed a cluster and showed the highest affinity to 2 Korean populations. In the phylogenetic tree Ryukyuans and Ainu were neighbors, but the genetic distance between them was larger than the distances between Ryukyuans and Hondo Japanese and between Ryukyuans and Korean populations. The geographic cline of the predominant haplotype in Ryukyuans, A*24 B*54-DRB1*0405, suggests that an ancestral population possessing A*24-B*54 DRB1*0405 moved into the Okinawa Islands after the divergence of Ryukyuans from the Ainu. Such a recent gene flow, probably from South China to the Okinawa Islands, is considered the major cause of difference in genetic characteristics between Ryukyuans and the Ainu. PMID- 10380373 TI - HLA DOA1 and DOB1 loci in Honduran women with cervical dysplasia and invasive cervical carcinoma and their relationship to human papillomavirus infection. AB - Molecular and epidemiological studies have demonstrated that certain types of human papillomavirus (HPV), mainly HPV-16 and HPV-18, are the primary causes of cervical cancer and its precursor lesions; there is now evidence for a clear association with specific HLA class I and class II loci contributing independently to the expression of cervical cancer. Among Honduran women carcinoma of the cervix is the most common type of cancer, and infections with high-risk HPV types are highly prevalent. To study the interactive role of viral host genetics, we performed PCR amplification of DNA and sequence-specific oligonucleotide probe typing on cervical scrapes from 49 women [24 with cervical intraepithelial neoplasia stage III or cervical cancer (severe cases) and 25 with stage I or II cervical intraepithelial neoplasia (mild cases)] and 75 control subjects to look for possible associations between HPV and HLA class II DQA1 and DQB1 alleles in the development of dysplasias and invasive cancer. This analysis revealed a predominance of HLA-DQA1*0301 among severe-case patients [relative risk (RR) = 3.45, p = 0.008), whereas DQA1*0501 was negatively associated (RR = 0.30, p = 0.03), suggesting a protective effect of this allele. HPV typing showed a decreased relative risk among the HPV-16 or HPV-18 carrying patients and other HPV-related positive patients in the presence of DQB1*0602 compared with positive control subjects (p = 0.04). No statistically significant allele frequency difference was observed between mild dysplasia cases and control subjects. The results suggest that DQA1*03011, which is in linkage desequilibrium with all HLA DR4 alleles, confers an increased risk for severe cervical dysplasia and invasive cancer, whereas DQA1*0501, which is in several DR52 haplotypes, has a protective effect. Furthermore, specific HLA-DQB1 sequences may be important in determining the immune response to HPV peptides and may affect the risk for cervical cancer after HPV infection in mestizo Honduran women. PMID- 10380375 TI - Patterns of gene flow inferred from genetic distances in the Mediterranean region. AB - The analysis of population structure may lead to inferences about demographic phenomena. In particular, regions of sharp genetic differentiation suggest the existence of factors that impaired gene flow and increased the evolutionary role of genetic drift. Here, we present an analysis of a data set of 10 allele frequencies in 39 populations of the Mediterranean region. As a preliminary step, we describe spatial patterns of allele frequencies using spatial autocorrelation analysis. We then construct a network connecting localities and estimate genetic distances along the edges of the network. By applying specific algorithms, we locate on the map the areas of sharpest genetic differentiation, or genetic boundaries. The main boundaries separate the northern and the southern coasts, especially in their western portions; in addition, several localities appear genetically isolated. The comparatively high genetic differentiation across the western Mediterranean, where the sea distances between localities are shorter, strongly suggests that the sea distance by itself can hardly be regarded as a major isolating factor among these populations. On the contrary, the decrease in genetic resemblance between populations of the 2 coasts as one proceeds westward may reflect an increased genetic exchange in the eastern Mediterranean basin or independent human dispersal along the 2 coasts or both. PMID- 10380374 TI - Effects of intragenic variability at 3 polymorphic sites of the apolipoprotein B gene on serum lipids and lipoproteins in a multiethnic Asian population. AB - We determined the allelic (X+/X-, M+/M-, and E+/E-) distribution frequencies of the XbaI, MspI, and EcoRI restriction fragment length polymorphisms (RFLPs) in the apolipoprotein B gene in a control group of 374 healthy Chinese, Malays, and Indians and in a hyperlipidemic cohort of 131 Chinese patients. Covariability between the RFLPs and serum lipid, lipoprotein, and apolipoprotein concentrations was also studied. We found a lower frequency (average 0.0829) of the X+ allele and higher frequencies of the E+ (average 0.9452) and M+ (average 0.9772) alleles in our study population compared with frequencies reported in other populations. The 3 polymorphic sites did not contribute to significant variations in lipid levels (p > 0.1 in all cases). Also, there was no significant variation in genotype frequencies between the control subjects and the hyperlipidemic subjects. Despite their relative close proximity within the APOB gene sequence, the 3 polymorphic sites did not show any significant linkage disequilibrium. However, the presence of the X+ cutting site was in linkage disequilibrium with the Del allele of the 5' insertion-deletion polymorphism and the E-allele was in linkage disequilibrium with the 3' VNTR located near the 3' end of the coding region of the APOB gene. PMID- 10380376 TI - Jamaican Symmetry Project: long-term study of fluctuating asymmetry in rural Jamaican children. AB - Fluctuating asymmetry, small deviations from perfect bilateral symmetry, is negatively correlated with health and positively correlated with sexual selection in human adults, but the accumulation, persistence, and fitness implications of asymmetries during childhood are largely unknown. Here, we introduce the Jamaican Symmetry Project, a long-term study of fluctuating asymmetry and its physical and behavioral correlates in rural Jamaican children. The project is based on an initial sample of 285 children (156 boys and 129 girls), aged 5 to 11 years. We describe the design of the project and the methodology of measuring 10 paired morphometric traits. All traits except hand width showed fluctuating asymmetry. Fluctuating asymmetries of the legs tended to be related and were less than half as great as fluctuating asymmetries of the arms and ears. Therefore the legs may show high developmental stability resulting from selection for mechanical efficiency. A fluctuating asymmetry composite score revealed that boys have significantly lower fluctuating asymmetry than girls and that this effect resides mainly in the elbows. There were significant positive relationships between composite fluctuating asymmetry and age, height, and weight, but multiple regression analyses showed that age was negatively related to fluctuating asymmetry, whereas body size was positively correlated. These findings are compared with results from recent English studies. PMID- 10380377 TI - Y-chromosome DNA haplotype 15 in Europe. AB - Haplotype 15 at 1 Y-chromosome-specific DNA polymorphism (p49/TaqI) was reported in a meta-analysis concerning 2418 males originating from 28 different geographic locations in Western Europe. The highest frequency of haplotype 15 (72.2%) was observed in French Basques, and it was previously deduced that this haplotype is the ancestral haplotype in Europe (Lucotte and Hazout 1996). Percentages of haplotype 15 geographic distribution show another high frequency in northwestern Europeans and a gradient of decreasing frequencies toward southeastern and peripheral countries. These results suggest that frequencies of haplotype 15 of the Y chromosome are useful to study the contribution of pre-Neolithic males to the present-day populations of Europe. PMID- 10380378 TI - Variation at 4 short tandem repeat loci in 8 population groups of India. AB - We have determined the nature and extent of variation at 4 STR loci (CSF1P0, TPOX, TH01, VWA) in 8 caste and tribal population groups of eastern and northern India. Large differences in allele frequencies among the groups were found. Average heterozygosities in all populations were high (approximately 80%). The overall extent of gene differentiation among the 8 groups was high (GST = 0.04). The nature of genomic affinities based on these 4 STR loci does not completely agree with our earlier finding based on classical genetic markers that geographic proximity of habitat has a greater influence on genetic similarity between populations than sociocultural proximity does. PMID- 10380380 TI - Need for NPs in developing countries. PMID- 10380379 TI - Problems with synthetic maps remain: reply to Rendine et al. PMID- 10380381 TI - Emphasis needed for primary prevention. PMID- 10380382 TI - Nursing in Greece in the 21st century. PMID- 10380383 TI - Supply and demand for nurses. PMID- 10380384 TI - Adults seeking presymptomatic gene testing for Huntington disease. AB - PURPOSE: To describe the expectations of those seeking presymptomatic gene testing for Huntington disease (HD). Identification of the gene for HD makes it possible to conduct testing to determine if a healthy person with a family history of HD has a mutation in this gene. Presymptomatic gene testing reveals the likelihood that a person will develop an inherited disease in the future. Understanding expectations allows for more complete assessment and counseling before presymptomatic gene testing for genetic diseases. DESIGN: Descriptive qualitative. The population was people with a family history of HD. The sample was 17 asymptomatic adults with a positive family history of HD who requested presymptomatic gene identification at one tertiary genetic counseling program, 1995 to 1996. METHODS: Semi-structured interviews concerning expectations of adults seeking presymptomatic genetic testing were conducted by telephone. Interviews occurred after the individuals had requested presymptomatic gene identification but before results were reported. Content analysis was used to identify the expectations and questions of those who had decided to seek presymptomatic testing. FINDINGS: Common expectations included anticipating relief from uncertainty, hoping to plan for their future health care and life decisions, wanting to know if their children were at risk of developing HD, anticipating loss of family support from relatives, expecting relief from self monitoring, venturing into the unknown, and planning for disclosure. Participants attempted to avoid their loss of genetic privacy by withholding the decision to seek testing from their primary care providers. CONCLUSIONS: Participants seeking presymptomatic HD gene testing consider the effect of gene identification on themselves and their families. A desire to limit insurance or employment discrimination contributes to subjects not seeking input from health care providers in their decision making. PMID- 10380385 TI - Effects of home care on caregivers' psychosocial status. AB - PURPOSE: To examine changes in the psychosocial status of caregivers of post surgical patients with cancer, and how their status was affected by (a) whether caregivers had physical problems of their own, and (b) whether the patient received a home care intervention. Many studies in this area to date have not included sufficient measurement points to identify fluctuations in psychosocial status over time. In addition, many have used caregiver health as an outcome rather than a predictor. DESIGN: Longitudinal, randomized trial using a sample of 161 caregivers of cancer patients being treated at one large university hospital in the northeastern United States, 1993-1996. Half the patients were randomly assigned to receive a standardized home-care nursing intervention. The population of interest was caregivers of patients who were (a) diagnosed with a solid-tumor cancer within the past 2 months, (b) age 60 or older, (c) hospitalized for surgical treatment of the cancer and expected to live at least 6 months, and (d) had a complex problem at hospital discharge. All caregivers were living with the patient at time of discharge. METHODS: Data were collected in structured interviews administered at the time of the patients' discharge and approximately 3 and 6 months later. Psychosocial status was measured using the Caregiver Reaction Assessment and the CES-Depression scale. A repeated-measures analysis of variance was performed for each psychosocial measure, using as factors Time (i.e., interview 1, 2, or 3), Group (treatment and control), and Caregiver Physical Problem. FINDINGS: Overall, psychosocial status improved from baseline to 3 months, and was about the same at 6 months. Among caregivers with physical problems, the psychosocial status of those in the treatment group declined compared to those in the control groups in the 3 months after discharge; an opposite pattern was observed in the following 3 months. CONCLUSIONS: People who are caregivers for cancer patients and have physical problems of their own are at risk for psychologic morbidity, which may have a delayed onset. This delay may reflect the replacement of an initial optimism with discouragement as the reality of long-term illness sets in. Home care may create a situation in which caregivers are required to confront the realities of long-term caregiving quickly, cutting short their initial optimism, but also preparing them for what is to come. PMID- 10380386 TI - Persistence of self in advanced Alzheimer's disease. AB - PURPOSE: To determine if evidence of the persistence of a sense of self or personal identity could be found in people in the middle and late stages of Alzheimer's disease. The theme of diminishing self pervades both the popular and professional literature on Alzheimer's disease. DESIGN: Qualitative using conversational analysis. The purposive sample was 23 residents of two urban nursing homes in the southeastern United States who were in the middle and late stages of Alzheimer's disease. Their mean Mini-Mental State examination score was 10.65. Nineteen subjects were women, four were men in this 1993-1997 study. METHODS: Analysis of 45 conversations lasting 30 minutes with nursing home residents with a diagnosis of probable Alzheimer's disease. Use of the first person indexical and other evidence, such as awareness and reactions to the changes that had taken place, in support of and counter to the notion of persistence of self, were sought in conversational analysis. FINDINGS: Respondents used the first person indexical frequently, freely, and coherently. Evidence was also present that participants were aware of their cognitive changes. Many struggled to provide an explanation, but none mentioned Alzheimer's disease. CONCLUSIONS: Evidence suggests the persistence of awareness of self into the middle and late stages of Alzheimer's disease. Failure to recognize the continuing awareness of self and the human experience of the person in the middle and late stages can lead to task-oriented care and low expectations for therapeutic interventions. The bafflement noted in respondents suggests that people should be told their diagnosis and offered an explanation of what this diagnosis means. PMID- 10380387 TI - Ethnomethodologic analysis of accounts of feeding demented residents in long-term care. AB - PURPOSE: To describe the self-organizing activities and the unspoken knowledge that nursing assistants employ in performing the work of feeding as a routine part of their activities. In U.S. long-term care facilities, activities related to medical and nursing care are frequently based on the ability of nursing assistants to successfully accomplish the work of feeding. Nurses are not always aware of the difficulties involved in feeding nor the means by which food is offered and intake calculated. DESIGN: Qualitative phenomenologic. METHODS: Observations were conducted during 12 weekday mealtimes, October and November 1994. Data were collected using participant observation of mealtimes in the congregate dining room of one licensed long-term care facility. Data were analyzed using the framework of ethnomethodology to indicate the methods nursing assistants used to make sense of their interactions with demented residents. FINDINGS: Nursing assistants used several common behaviors: grouping the "difficult feeders" together; "marking the borders" by maintaining a tidy tray; and "loading the spoon" to maximize the amount of food given at one time. Estimations of the quantity of intake varied according to ideas about the nutritional content of various foods. CONCLUSIONS: Unsuccessful feeding interactions have implications for the quality of life for the demented. When inadequate intake leads to significant weight loss, medical and nursing interventions should be implemented. PMID- 10380388 TI - Scenario--long-term care for the elderly. PMID- 10380389 TI - Sleep patterns of sheltered battered women. AB - PURPOSE: To describe sheltered battered women's sleep patterns and resulting daytime fatigue. The model for symptom management framed this study to describe one component of the model--symptom experience. Beginning evidence suggests that sheltered battered women experience disturbed sleep and fatigue that can interfere with vital activities. DESIGN: Descriptive using a convenience sample of 50 ethnically diverse women residing at least 21 days in battered women's shelters located in one western U.S. city. The study was done in 1997. METHODS: The Pittsburgh Sleep Quality Index (PSQI), the Mini Motionlogger Actigraph, a sleep diary, the Visual Analogue Scale for Fatigue, and an open-ended interview were used to collect data. Descriptive and inferential statistics were used to analyze the quantitative data. Qualitative data were analyzed to determine patterns and themes. FINDINGS: Seventy percent of the women had global PSQI scores of greater than five indicating poor sleep and 17 (34%) had a sleep efficiency index of 80% or less. Fourteen (28%) of the women went to bed very fatigued (> 66 mm) and 20 (40%) woke up very fatigued (> 33 mm). CONCLUSIONS: The majority of sheltered battered women experienced disturbed sleep and daytime fatigue. Both personal and environmental variables were found to significantly affect sleep patterns. Sheltered battered women can benefit from information about sleep disturbances and sleep enhancing self-care strategies. PMID- 10380390 TI - Linking concepts of enduring, uncertainty, suffering, and hope. PMID- 10380391 TI - A model for cognitive-behavioral interventions in cancer pain management. AB - PURPOSE: To propose a model for predicting success with cognitive-behavioral interventions in cancer pain management. Practice guidelines are useful, however nurses currently have little theoretic or empiric basis for choosing one particular strategy over another. Moreover, nurses have no way of knowing if a particular intervention is likely to work. ORGANIZING CONSTRUCT: The model indicates characteristics of a person in relation to interventions including skill and ability, outcome expectancies, perceived credibility, history of use, preferred coping style, and pain outcomes. SOURCES: The model was developed using sources identified through a literature search of relevant topics in MEDLINE, CINAHL, and Psychlit (1996-1997), as well as through clinical experience. CONCLUSIONS: Continued empiric testing of the model is necessary to confirm proposed relationships and to assess accuracy of the model's predictions with various cognitive-behavioral interventions. With this testing, the model can help nurses select appropriate interventions for individual patients. PMID- 10380392 TI - Changes in the nurse workforce. PMID- 10380393 TI - Influence of managed care on professional nursing practice. AB - PURPOSE: To explore how nurses in one U.S. state perceived that managed care influenced professional nursing in that state. The nursing community is challenged to move with haste in demonstrating, through research, the clinical and economic value that nurses add to cost-effective outcomes. DESIGN: A Delphi survey in 1996 of a convenience sample of 84 clinical nurse specialists (CNSs) and nurse practitioners (NPs) in California. METHODS: CNSs and NPs contributed to the list of managed care influences on nursing practice. fifty-seven (68%) completed the third and final round. FINDINGS: Panelist agreement was the highest for (a) exploring new approaches to providing quality care more cost-effectively, (b) expanding nurse practitioners' role in primary care, and (c) more effectively partnering with clients in helping them assume greater self-responsibility for their health. Greatest threats were perceived to be hassles involved in seeking authorization for care and responding to payment denials; the tenuous job market for nurses; and encroachment on nursing practice by others. CONCLUSIONS: The findings can assist nurses in states with low managed-care concentration to create their preferred future within health care delivery. A more highly educated nurse workforce will be needed for 21st century health systems in which more care is likely to be delivered outside hospitals. PMID- 10380394 TI - Reimbursement to advanced practice nurses (APNs) through Medicare. AB - PURPOSE: To analyze why U.S. legislation for direct Medicare reimbursement to advanced practice nurses (APNs) was approved during the 105th congressional session. Help in understanding the complexities of policy making and in strategizing future policy activities is needed. Given their vast numbers, nurses are a formidable group with potential to influence policy. ORGANIZING CONSTRUCT: Kingdon's (1995) framework indicates that policy formation involves constant interaction among participants and the problem, its politics, and policy. When a policy is enacted into legislation, the three come together and an opportunity for policy formation exists, if even for a short period. Methods for data extraction included telephone interviews, information provided by the American Nurses Association (ANA), and information from the internet to access information. FINDINGS: Lack of access to health care by Medicare beneficiaries, tagging a policy proposal onto the Balanced Budget Act of 1997, the national mood to provide cost-effective quality care, and lobbying by interest groups helped to make direct Medicare reimbursement for advanced practice nurses a reality. CONCLUSIONS: When lobbying for the enactment of legislation, nurses should decrease their tendency to fragment. Nurses should be more effectively organized and concentrate on specific agendas. Working together can influence policy and change clinical situations to improve patient care. PMID- 10380395 TI - Proliferation of non-physician providers as reported in the Journal of the American Medical Association (JAMA), 1998. PMID- 10380396 TI - Slow growth in the United States of the number of minorities in the RN workforce. AB - PURPOSE: To assess the extent to which the number of minority RNs has grown during the past 20 years, and to identify and compare key trends in personal and professional characteristics among minority groups and between minority and majority populations of RNs. Nursing education programs, employers, philanthropic organizations, and governments have expended considerable effort and resources to increase the number of minorities in nursing. DESIGN: Longitudinal analysis of trends in the number, education, employment, and earnings of minority RNs from 1977 to 1997. METHODS: Descriptive analysis of data from the U.S. National Sample Surveys of the Population of Registered Nurses, 1977-1996; and data from the U.S. Bureau of the Census Current Population Survey (CPS) Outgoing Rotation Group Annual Merged Files, 1977-1997. FINDINGS: In the past 20 years, the number of minority RNs has grown from 87,386 (or 6.3% of the total supply of RNs) in 1977 to 246,364 RNs (9.7%) in 1996. The number of Black (nonHispanic), Asian Pacific/Islanders, and American Indian/Alaskan Native nurses nearly tripled in this period while the number of Hispancis doubled. Although these rates of growth are impressive, the percentage of minorities in nursing lags considerably behind the percentage (18.3%) who are teachers, and the percentage (28.2%) in the U.S. population. CONCLUSIONS: Studies are needed to determine the barriers that exist in nursing education programs, health care organizations, and society in general that deter minorities from a nursing career. Without this understanding, efforts to design and implement ideas to attract, educate, and retain minorities in nursing education and later in the workplace are hampered. PMID- 10380397 TI - Interview of visiting health administrators from the Ministry of Health, Beijing, China. Interview by Wen-Yin Chang. PMID- 10380398 TI - Scenario--nursing and nursing education. PMID- 10380399 TI - The classification of smile patterns. AB - Although "smile therapy" is still in its infancy, society has already placed a great demand on dentists to evaluate and treat smiles. The smile classification scheme and vocabulary presented in this article will aid in discussions between patient and dentist regarding esthetic treatment. PMID- 10380400 TI - A prescription for the successful use of heavy filled composites in the posterior dentition. PMID- 10380401 TI - Continuing education and dentistry. PMID- 10380402 TI - Taking control of your practice. PMID- 10380403 TI - Diagnostic provisional restorations in restorative dentistry: the blueprint for success. AB - There is no question that patients today demand a sophisticated level of restorative dentistry, in terms of both esthetics and function. No elective restorative dentistry should be undertaken without a clear understanding of the patient's expectations and the limitations of restorative therapy. The dentist should have a clear picture in mind of the final results before initiating irreversible therapy. The use of mounted diagnostic casts, diagnostic wax-ups and provisional restorations permits patient acceptance to be obtained before the definitive phase is initiated. Too often the dentist does not take advantage of this important restorative option, with disastrous results when definitive restorations are viewed by the patient for the first time. By following the plan of treatment outlined in this article, such disasters can be avoided. PMID- 10380404 TI - Primary cleft lip and palate. PMID- 10380405 TI - Preparing for medical emergencies in the dental office. AB - If you discover an unconscious patient in your office, attend to the ABCs while you evaluate the patient's medical history and piece together the events leading up to the emergency. These actions will help you arrive at a diagnosis. Then as the emergency cart and team arrive, you will be able to provide good, safe care to stabilize the patient and get him or her to a medical facility. PMID- 10380406 TI - Practical issues in delivering geriatric dental care. PMID- 10380407 TI - When the unthinkable happens: post-exposure prophylaxis of HIV-contaminated percutaneous injuries. PMID- 10380408 TI - [The family, spare-time activities and environmental health risks]. PMID- 10380409 TI - [Preservatives in nebulizer solutions: risks without benefit]. PMID- 10380410 TI - [Reactions and interactions of drugs. Safety measures from Drug Safety Committee of the United Kingdom]. PMID- 10380411 TI - BM 144: an original thromboxane A2 receptor antagonist derived from torasemide. AB - Torasemide, a new sulfonylurea high ceiling loop diuretic, has been demonstrated to induce a concentration dependent relaxation of canine coronary precontracted with thromboxane A2 (TXA2). With the aim to develop more potent TXA2 receptor antagonists, we investigated a series of torasemide derivatives. This pharmacomodulation led to the discovery of a sulfonyl-cyanoguanidine (BM 144) which presents the same pharmacological profile as sulotroban, a TXA2 receptor antagonist used as reference. PMID- 10380412 TI - [The first vaccine for control of cervical cancer]. PMID- 10380413 TI - Alternatives to the sequential lineup: the importance of controlling the pictures. AB - Because sequential lineups reduce false-positive choices, their use has been recommended (R. C. L. Lindsay, 1999; R. C. L. Lindsay & G. L. Wells, 1985). Blind testing is included in the recommended procedures. Police, concerned about blind testing, devised alternative procedures, including self-administered sequential lineups, to reduce use of relative judgments (G. L. Wells, 1984) while permitting the investigating officer to conduct the procedure. Identification data from undergraduates exposed to a staged crime (N = 165) demonstrated that 4 alternative identification procedures tested were less effective than the original sequential lineup. Allowing witnesses to control the photographs resulted in higher rates of false-positive identification. Self-reports of using relative judgments were shown to be postdictive of decision accuracy. PMID- 10380414 TI - Applicant reactions to test score banding in entry-level and promotional contexts. AB - This series of field studies used a fairness framework to investigate applicant reactions to test score banding in 3 police selection contexts. Studies 1 (N = 85) and 2 (N = 369) involved applicants for entry-level positions, and Study 3 (N = 39) involved applicants for promotion. Across all 3 studies, race interacted with applicants' belief that banding is associated with affirmative action to affect measures of fairness and organizational outcomes such as attractiveness and perceived employee relations. Reactions were also related to applicants' perceived outcomes as a result of banding. Results are explained in terms of self interest and suggest that reactions to banding are largely a function of the association of banding with affirmative action. PMID- 10380415 TI - The effect of the cognitive interview on face identification accuracy: release from verbal overshadowing. AB - Three experiments tested the effect of verbal description on face identification accuracy. Based on verbal overshadowing research, it was predicted that enhancing verbal description of a face would reduce subsequent face identification accuracy. Experiment 1 tested and confirmed this hypothesis using the cognitive interview to enhance verbal description; face identification accuracy was reduced significantly following the cognitive interview, compared with a standard police interview. Experiments 2 and 3 tested and confirmed the hypothesis that verbal overshadowing would be reduced when a delay is inserted between verbal description and face identification, hence resulting in "release from verbal overshadowing." These results suggest that in the verbal overshadowing task, the verbal description does not overwrite the visually based representation of the face in memory but rather makes it less accessible at the time of face identification. The cognitive interview reduces face identification accuracy only when the identification follows description immediately--a rare situation in real criminal cases. PMID- 10380416 TI - Efficacy beliefs as a moderator of the impact of work-related stressors: a multilevel study. AB - This study built on previous exploratory research (S. M. Jex & D. M. Gudanowski, 1992) that examined both self-efficacy and collective efficacy as moderators of stressor-strain relations. Based on survey data collected from 2,273 U.S. Army soldiers representing 36 companies, it was found that both self- and collective efficacy moderated the relationship between stressors and strains. Multilevel random coefficient model results revealed that respondents with strong self efficacy reacted less negatively in terms of psychological and physical strain to long work hours and work overload than did those reporting low levels of efficacy. In addition, respondents with high levels of self-efficacy responded more positively in terms of job satisfaction to tasks with high significance than did those with low efficacy. The results also revealed that group-level collective efficacy moderated the relationship between work overload and job satisfaction and between task significance and organizational commitment. Limitations of the study and implications of these findings are discussed. PMID- 10380417 TI - Reasoning about scientific evidence: effects of juror gender and evidence quality on juror decisions in a hostile work environment case. AB - This study examined whether participants were sensitive to variations in the quality of an experiment discussed by an expert witness and whether they used heuristic cues when evaluating the expert evidence. In the context of a hostile work environment case, different versions of the expert testimony varied the presence of heuristic cues (i.e., whether the expert's research was generally accepted or ecologically valid) and evidence quality (i.e., the construct validity of the expert's research). Men who heard expert testimony were more likely to find that the plaintiff's workplace was hostile than were men who did not hear the expert testimony; expert testimony did not influence women's liability judgments. Heuristic cues influenced participant evaluations of the expert testimony validity, but evidence quality did not. Cross-examination did not increase juror sensitivity to evidence quality. Implications for science in the legal system are discussed. PMID- 10380418 TI - Team effectiveness: beyond skills and cognitive ability. AB - On the basis of job analysis results, the validity of using measures of general cognitive ability, job-specific skills, and personality traits jointly at both the individual level and the group level to predict the performance of 79 four person, human resource work teams was evaluated. Team member trait and job skill scores were aggregated with a conjunctive model of task performance. At the individual level of analysis, measures of personality (i.e., Agreeableness and Conscientiousness) predicted peer ratings of team member performance beyond measures of job-specific skills and general cognitive ability. Similarly, at the group level of analysis, both Agreeableness and Conscientiousness predicted supervisor ratings of work team performance, objective measures of work team accuracy, and work completed. At both the individual and group levels, the trait of Agreeableness predicted Interpersonal Skills. PMID- 10380419 TI - Outcomes of self-labeling sexual harassment. AB - Research has consistently documented a discrepancy between experiencing offensive sex-related behaviors and labeling these situations as sexual harassment, leading to several attempts to understand this phenomenon. The authors propose that the issue of why it is that women who report such experiences generally do not indicate that they have been sexually harassed is an important psychological question, and may provide a path through the nested meanings of workplace harassment. The authors argue for the value of moving beyond a descriptive approach to this issue by examining the effects of self-labeling on the psychological, health, and work-related outcomes of unwelcome, sex-related experiences. They present data from female employees working in 3 separate organizations, demonstrating that women exposed to such behaviors report very similar consequences, whether they label their experiences as harassment or not. PMID- 10380420 TI - Converting the unconverted: the effect of inclination and opportunity to discount health-related fear appeals. AB - Two experiments indicated that the conventional wisdom for designing fear appeals, higher fear arousal, and a consequences-recommendations ordering, was more persuasive for adherents, or those who were already following the advocated recommendations. Instead, lowering the level of fear arousal and reversing the order of the consequences and recommendations were more effective for persuading the unconverted. The unconverted were more persuaded by the latter message format because it reduced the level of message discounting. Specifically, unconverted participants who received either a low fear appeal or recommendations preceding consequences perceived themselves to be more susceptible, perceived the consequences as more severe, regarded the recommendations as more efficacious, believed they were more able to follow the recommendations, and were less likely to refute the message claims. PMID- 10380421 TI - Proactive personality and career success. AB - This study examined the relationship between proactive personality and career success by surveying a sample of 496 employees (320 men and 176 women) from a diverse set of occupations and organizations. Proactive personality was positively associated with both self-reported objective (salary and promotions) and subjective (career satisfaction) indicators of career success. Hierarchical regression analyses showed that proactive personality explained additional variance in both objective and subjective career success even after controlling for several relevant variables (demographic, human capital, motivational, organizational, and industry) that have previously been found to be predictive of career outcomes. These findings were consistent using both self-report and significant--other ratings of proactive personality. PMID- 10380422 TI - The effect of leader outcomes on influence, attributions, and perceptions of charisma. AB - This study addressed how various outcomes to a leader might influence not only how that leader is perceived but also the degree of influence that leader might subsequently obtain. On the basis of recent charismatic leadership theories, it was expected that leaders who appeared willing to endure hardship for the expression of their beliefs would be perceived differently than leaders who appeared to benefit in some way. The relationship between outcomes and subsequent leader influence was confirmed. Sacrificing resulted in greater influence, whereas benefiting reduced it. This relationship was mediated by attributions made about leader motives. The relationship between outcomes and influence was also mediated by perceptions of charisma. PMID- 10380423 TI - Parents' job insecurity affects children's academic performance through cognitive difficulties. AB - The authors developed and tested a model in which children who perceive their parents to be insecure about their jobs are distracted cognitively, which in turn affects their academic performance negatively. Participants were 102 female and 18 male undergraduates (mean age = 18 years), their fathers (mean age = 49 years), and their mothers (mean age = 47 years). Students completed questionnaires measuring perceived parental job insecurity, identification with parents, and cognitive difficulties; 3 months later, they also reported their midyear grades. Fathers and mothers each completed questionnaires assessing their job insecurity. Support for the model was obtained using LISREL 8, and as predicted, children's identification with their mothers and fathers moderated the relationship between their perceptions of their mothers' and fathers' job insecurity and their own cognitive difficulties. PMID- 10380424 TI - The effects of interpersonal trust on work group performance. AB - This study explored 2 questions: Does the level of trust within a group affect group performance? If so, how does this relationship operate? An experimental method was used to examine 2 roles through which interpersonal trust could affect group performance: a main effect and a moderating effect. The data do not support the main effect that has dominated the literature on interpersonal trust. The data do support the moderating role: Trust seems to influence how motivation is converted into work group processes and performance. On the basis of these findings, it is suggested that trust may be best understood as a construct that influences group performance indirectly by channeling group members' energy toward reaching alternative goals. PMID- 10380425 TI - Production uncertainty as a contextual moderator of employee reactions to job design. AB - A number of authors in the job design field have proposed production uncertainty, the degree to which a qualified incumbent faces unexpected problems in the course of job performance, as a possible moderator of the effectiveness of job design. However, empirical support for this view is limited and has not been explicitly recognized within mainstream job design theory. This study of production operators in a waste water treatment setting provides further empirical support for production uncertainty as a contextual variable influencing job design outcomes, demonstrating that the relationship between job control and affective outcomes (job satisfaction and intrinsic motivation) varies with the level of production uncertainty. PMID- 10380426 TI - The correlation of the metastatic ability with keratin expression in cultured murine melanoma cell lines, B16-F1 and-F10. AB - Keratin is an intermediate filament that is a major structural protein of epithelial cells. Until now, the expression of keratin in melanoma cells has not been well understood. Recently, it has been reported that keratin expression is correlated with invasive and metastatic behavior in a variety of cell types. We report keratin expression in cultured murine melanoma cell lines B16-F1 (low incidence of lung colonization) and F10 (high incidence of lung colonization) using an aqueous solution (10 mM Tris-HCl (pH 7.4)/10 mM EDTA/phenylmethyl sulphonyl fluoride (PMSF, 10 micrograms/ml). By comparing these two cell lines, we investigated whether differences in keratin expression can influence the metastatic ability of tumor cell lines in vitro. However, no remarkable differences in keratin expression were found in these cell lines. PMID- 10380427 TI - Identification of several clinical isolates of dermatophytes based on the nucleotide sequence of internal transcribed spacer 1 (ITS 1) in nuclear ribosomal DNA. AB - Nucleotide sequences of internal transcribed spacer 1 (ITS 1) in nuclear ribosomal DNA from seven morphologically unidentified dermatophyte isolates were determined. The sequences were compared with those of typical isolates of Trichophyton (T.) mentagrophytes var. interdigitale, T. rubrum, and Epidermophyton floccosum. Two of the isolates were classified as T. rubrum and the other five as T. mentagrophytes var. interdigitale. The results did not conflict with identifications using other molecular techniques, including random amplification of polymorphic DNA (RAPD) analysis and restriction enzyme analysis of mitochondrial DNAs. Thus, the nuclotide sequence of ITS 1 is possibly a good molecular marker for identification of these major anthropophilic dermatophyte species. PMID- 10380428 TI - An assessment of the role of Candida albicans antigen in atopic dermatitis. AB - An immediate hypersensitivity reaction to Candida albicans (C. albicans) antigen has been observed in patients with atopic dermatitis. Recent data from a comparative study of the immune response to C. albicans antigen in patients with atopic dermatitis and non-atopics suggest a shift form type 1 helper T cells to type 2 helper T cells in the immune response to C. albicans antigen in atopic dermatitis. To delineate the role of C. albicans in the pathogenesis in atopic dermatitis, we evaluated skin reaction of C. albicans antigen, as well as the serum IgE antibody level against C. albicans in patients with atopic dermatitis, patients with nasal allergy, and non-atopics. In addition, the clinical effect of antifungal drugs was evaluated in the patients with atopic dermatitis. As a result, we found that immediate hypersensitivity to C. albicans antigen is strongly correlated with the patients with atopic dermatitis. On the other hand, the delayed-type hypersensitivity to this antigen, which is highly prevalent in atopics without dermatitis as well as non-atopics, was reduced in most of these patients. Antifungal drugs markedly improved the skin manifestations in patients with atopic dermatitis that have IgE antibodies against C. albicans, and the serum IgE levels also decreased. These results suggest that C. albicans antigen is a potent intrinsic factor in inducing skin lesions in atopic dermatitis because of IgE-mediated hypersensitivity of C. albicans antigen. PMID- 10380429 TI - Clinico-bacteriological study of pyodermas in children. AB - One hundred cases of pyodermas in children were investigated clinically and bacteriologically. Nasal and throat swabs from all cases were subjected to bacteriological examination. Most of the children (42%) were in the 1-4 year age group. The majority (58%) belonged to lower socio-economic groups with poor standards of hygiene. A history of over-crowding was obtained from 87% of cases, 82% were undernourished. Most of the children (68%) reported during the hot and humid months of June, July, August, and September. Primary pyodermas were observed in 72% of the children, and secondary pyodermas in 28%. Impetigo was the commonest primary pyoderma (48.61%); among secondary pyodermas, infected scabies was noted predominantly (42.86%). The face and legs were more commonly involved. Bacteriological cultures from pyoderma lesions revealed a single microorganism in the majority of the patients (84%). Staphylococcus aureus was isolated in pure culture from 48% and pure beta-hemolytic streptococci from 36%. A combination of both was obtained from 16%. No other organism was isolated from any case. A similar pattern was also observed in cultures from the nose and throat. Only 46 out of the 64 strains (84.3%) of Staphylococcus aureus isolated from pyoderma were typable. The majority (39.1%) showed a mixed pattern of phages; the second commonest was the non-allocated phage type (30.4%). Nasal flora had more of the non-allocated phage type (50%); two out of the three strains (66.6%) isolated from the throat showed a mixed pattern. All the strains of beta hemolytic streptococci, isolated either from lesions of pyoderma, nose, or throat belonged to group A. Staphylococcus aureus and showed a high sensitivity to netilmycin (100%), ofloxacin (98.4%), amoxycillin/clavulanic acid (96.9%), ciprofloxacin (89.1%) and gentamycin (84.4%) but a high resistance to penicillin (85.9%). A greater correlation was noted between nasal flora and organisms causing pyodermas. A change in the pattern of organisms causing pyodermas in children and their antibiotic sensitivities in this part of the globe has been observed in this study. The role of endogenous nasal and throat flora in the causation of pyodermas has also been highlighted. PMID- 10380430 TI - The spectrum of mucocutaneous manifestations during the evolutionary phases of HIV disease: an emerging Indian scenario. AB - Third world countries, including India, lack sophisticated investigations to assess the progression of HIV disease. Hence, this study was undertaken to determine the clinical mucocutaneous markers of HIV disease and to establish its relationship with the stage of the disease. This was an observational institutional study of 75 patients with mucocutaneous disorders and HIV infection recruited over a period extending from September of 1996 to June of 1998. The patients with mucocutaneous lesions were staged according to the Centers for Disease Control classification system for HIV infection (1986). The most frequent mode of acquisition of HIV infection was heterosexual contact (96%). The patients were broadly categorized into two groups. The AIDS group was comprised of patients who were in group IV and the early HIV infection group included patients in group II and III; none were detected in group I. Forty-eight cases belonged to the AIDS group, and 27 belonged to the early HIV infection group. A total of 207 dermatoses were diagnosed and grouped as fungal, viral, bacterial, or miscellaneous. The common mucocutaneous disorders in order of frequency observed in this study were: candidiasis, dermatophytosis, herpes simplex, oral aphthae, xerosis/ichthyosis, scabies, HPV infection, molluscum contagiosum, and psoriasis. Xerosis/acquired ichthyosis and giant molluscum contagiosum were characteristically seen in group IV of HIV disease, whereas oral candidiasis, oral aphthae, papular dermatitis of HIV, and psoriasis were early warning signs. The mean number of dermatoses per patient in group IV was 3.15; in group III, it was 2.41; and in group II, it was 1.5. There was a statistically significant difference between the early HIV infection group and AIDS group with regard to number of dermatoses. Apart from syphilis and human papilloma virus infection, the treatment outcomes were satisfactory. PMID- 10380431 TI - Macular amyloidosis: etiological factors. AB - Macular amyloidosis is the commonest form of primary localized cutaneous amyloidosis; its etiology remains unclear. Various incriminating factors include genetics, sunlight, and friction. Seventy-five patients with a clinical diagnosis of macular amyloidosis (confirmed by at least two observers) were enrolled in the study. A detailed history was elicited in relation to the etiological factors and followed by cutaneous examination. Skin biopsy was performed in 44 patients. Amyloid deposits were demonstrated in 21 (48%) of the biopsies. The female to male ratio was over 3:1. Patients with skin type V had a more delayed onset than those with skin types III and IV. Two patients gave histories of regularly subjecting their skin to friction. There was no correlation between the sites affected and the sites subjected to friction, hair style of the patient, or the use of cosmetics. Family history was present in 7 (9.3%) patients. A majority, 44 (58.7%) of the patients, had involvement of both sun-exposed and sun-protected sites. To conclude, there was no direct correlation between macular amyloidosis and atopy, sunlight, or friction to the skin in our patients. Multiple factors may play a collective role in the genesis of macular amyloidosis to variable degrees. These include racial, familial, and environmental factors, atopy, sunlight, friction, and female gender. PMID- 10380432 TI - A case of metastatic extramammary Paget's disease that responded to combination chemotherapy. AB - Extramammary Paget's disease is considered to be a malignant tumor originating from the sweat glands. Some cases of extramammary Paget's disease infiltrate the dermis and metastasize to the regional lymph nodes. No standard treatment has been established for advanced cases. Few previous studies of the treatment for metastatic Paget's disease have revealed an effective regimen. The prognosis of metastasized cases is very poor. We encountered a patient in whom extramammary Paget's disease had metastasized to the lymph nodes beyond the regional lymph nodes and systemic chemotherapy was partially effective. Combination chemotherapy consisted of mitomycin C 3.5 mg/m2 and epirubicin 50 mg/m2 on day 1, vincristine 0.6 mg/m2 on days 1 and 7, cisplatin 30 mg/m2 from days 1 to 3, and 5 fluorouracil 350 mg/m2 from days 3 to 7. After two courses of chemotherapy, the metastatic lymph nodes decreased in size by more than 90% compared to that before chemotherapy. We defined the treatment effect of this regimen as a partial response (PR). Microscopic examination of the resected lymph nodes revealed replacement of metastatic lesions by fibrous tissue, suggesting a therapeutic effect. Anorexia, alopecia, and leukopenia (neutropenia) have been reported as toxicities, but all were tolerated. Our results may provide useful indications for the management of this tumor. This particular combination chemotherapy is recommended for extramammary Paget's disease patients with systemic nodal metastases. PMID- 10380433 TI - Sweat gland adenoma with predominant myoepithelial differentiated features: case report and immunohistochemical study. AB - A 56-year-old man presented with a subcutaneous mass adjacent to the sternoclavicular joint region, which had predominant spindle-shaped myoepithelial cells and a small number of tubular or trabecular epithelial cells that resembled eccrine differentiation under a light microscope. Immunohistochemical studies showed positive staining for muscle-specific actin and vimentin in the myoepithelial cells, but staining was negative for S-100 protein and keratin. The tubular and trabecular epithelial cells stained positively for keratin, EMA and CEA. We believe that this rare tumor had the potential for biphasic differentiation and should be differentiated from other biphasic differentiated or mixed tumors in this site. Immunohistochemical staining was useful for diagnosis and differential diagnosis. PMID- 10380434 TI - Progeria (Hutchinson-Gilford): a case report. AB - A new case with the typical features of progeria (Hutchinson-Gilford) occurred in India. Histopathology of the skin showed atrophic epidermis and diffuse fibrosis of dermis with loss of appendages. Roentgenographic findings were characteristic of progeria. The child also had a gangrenous ulcer over the left foot, a finding not highlighted in the literature. PMID- 10380435 TI - Pseudolymphoma syndrome due to carbamazepine. AB - We described a 29-year-old Japanese woman with a pseudolymphoma syndrome due to carbamazepine. Physical examination revealed a diffuse erythematous papular eruption over almost all of her body with generalized lymphadenopathy. Histopathologic examination disclosed histology compatible with the nodular pattern of pseudo-T-cell lymphoma. The Southern blot analysis did not disclose the monoclonality of the gamma T cell recepter gene. We consider that it is very important to recognize this pseudolymphoma syndrome due to carbamazepine to avoid possibly subjecting the patient to unnecessary chemotherapy. PMID- 10380436 TI - Aggregated dilated pores. AB - We report a 74-year-old Japanese man who had a 10-year history of approximately 20 open comedones crowded onto his lateral neck. An excisional biopsy revealed that each comedo was histologically a dilated pore. Dilated pores are usually solitary. The multiple and aggregated dilated pores seen in our case have never been reported. PMID- 10380438 TI - Complementary and alternative therapies in women's health. A home study program sponsored by the Journal of Nurse-Midwifery. PMID- 10380437 TI - Annular vasculitis in association with sarcoidosis. AB - We report a case of cutaneous vasculitis with annular features in association with sarcoidosis. A 36-year-old woman presented with fever, polyarthralgias, erythema nodosum, bilateral hilar lymphadenopathy, and induration of a long standing scar on the face. In addition, she developed annular, erythematous, and purpuric patches over her thighs and buttocks that were histologically characterized by a small vessel leukocytoclastic vasculitis. The presence of circulating immune complexes in the early stages of sarcoidosis might be related to the occurrence of the vascular damage. PMID- 10380439 TI - Complementary and alternative healing in midwifery care. PMID- 10380440 TI - Complementary and alternative medicine in women's health. Developing a research agenda. AB - Complementary and alternative medicine is becoming an established intervention modality within the contemporary health care system. Various forms of complementary and alternative medicine are used by patients and practitioners alike, including chiropractic, massage, botanical medicine, homeopathy, and energy therapies. The National Center for Complementary and Alternative Medicine was established within the National Institutes of Health to facilitate evaluation of these alternative therapies, establish an information clearinghouse, and promote research in the field. This article discusses several aspects of complementary and alternative medicine, relates them to women's health, and describes the need for a research agenda to evaluate the impact of the complementary and alternative medicine modalities used for important conditions affecting women. PMID- 10380441 TI - A national survey of herbal preparation use by nurse-midwives for labor stimulation. Review of the literature and recommendations for practice. AB - To document the use of herbal preparations for cervical ripening, induction, and augmentation of labor by certified nurse-midwives (CNMs) and nurse-midwifery education programs, a national survey of 500 members of the American College of Nurse-Midwives was conducted. Forty eight nurse-midwifery education programs were also surveyed to determine whether they were formally or informally educating students in the use of herbal preparations for cervical ripening, induction, or augmentation of labor. The results of this study, a review of the literature, professional issues, and recommendations for clinical practice are presented in this article. Of 500 questionnaires mailed to ACNM members, 90 were returned from CNMs who used herbal preparations to stimulate labor and 82 were returned from CNMs who did not use herbal preparations to stimulate labor. Three questionnaires were excluded due to incomplete data or blank questionnaires. No significant differences were noted in relations to geographical region, midwifery education, or highest level of education between the CNM respondents who did and those who did not use alternative methods to stimulate labor. Of the CNMs who used herbal preparations to stimulate labor, 64% used blue cohosh, 45% used black cohosh, 63% used red raspberry leaf, 93% used castor oil, and 60% used evening primrose oil. CNMs who used herbal preparations to stimulate labor were younger (43 versus 45 years, P < .01) and more likely to deliver at home or in an in-hospital or out-of hospital birthing center (P < .0006), than CNMs who never used herbal preparations to stimulate labor. The most cited reason for using herbal preparations to stimulate labor was that they are "natural," whereas the most common reason for not using herbal preparations was the lack of research or experience with the safety of these substances. Sixty-nine percent of CNMs who used herbal preparations to stimulate labor learned about using them from other CNMs, 4% from formal research publications, and none from their formal education programs. Although 78% of the CNMs who used herbal preparations to stimulate labor directly prescribed them and 70% indirectly suggested them to clients, only 22% had included them within their written practice protocols. Seventy-five percent of the CNMs who used herbal preparations to stimulate labor used them first or instead of pitocin. Twenty-one percent reported complications including precipitous labor, tetanic uterine contractions, nausea, and vomiting. Sixty-four percent of the nurse-midwifery education programs included instruction in the use of herbal preparations to stimulate labor in their formal curricula, and 92% included informal discussions on the use of herbal preparations. Evening primrose oil was the most common herbal preparation discussed in nurse-midwifery education programs. Castor oil was the most commonly used herbal preparation used by nurse midwives in clinical practice. PMID- 10380443 TI - Herbs and the childbearing woman. Guidelines for midwives. AB - The use of herbs to promote health or treat disease has become popular, and midwives increasingly encounter questions from childbearing clients regarding herbs. This article provides an overview of key concepts regarding the incorporation of herbs into clinical practice and discusses the preparation and administration of herbal treatments for common concerns of pregnancy. Safety issues are emphasized throughout. PMID- 10380442 TI - Acupuncture and acupressure. Applications to women's reproductive health care. AB - An introduction to the therapeutic applications, history, and theory of acupuncture and acupressure is presented. The traditional concepts that underlie treatment of imbalances of ch'i, or vital energy, are presented, along with the theories of yin and yang, meridians, vital substances, pathogenic factors, five phases, and the eight principle patterns. Contemporary Western research findings on the biochemical mediaries and effects of acupuncture are reviewed. Clinical applications to women's reproductive care that are presented include treatment for dysmenorrhea, infertility, and childbearing. Data on clinical trials are reviewed, and licensure and educational preparation for practice of these modalities are discussed. PMID- 10380444 TI - Introducing herbal medicine into conventional health care settings. AB - Herbal therapy is one of several holistic therapies gaining recognition within the health care community in the United States. As a discipline, herbal medicine is in its infancy regarding educational standards for credentialling, standardization, and regulation of products and clinical applications within this health care system. This article discusses professional considerations for midwives who are interested in integrating herbal healing into their clinical practices, and offers examples of how to incorporate herbal medicine into midwifery care. Resources for practitioners including books, newsletters, journals, courses, computer sites, and databases are presented. The author offers guidance for creating an herbal practice manual for the midwifery office as well as the hospital setting and for documenting herbal healing in the medical record. Collegial support, barriers to practice, liability, and insurance issues are discussed. A clinical applications section includes specific herbal formulas for preconception health, pregnancy-induced hypertension, gestational diabetes, and postdates pregnancy. PMID- 10380445 TI - Menopause and beyond. The wise woman way. AB - The Wise Woman tradition seeks to heal the whole individual. The primary techniques involve nourishing the woman through storytelling, simple ceremony, and dietary herbs. The "mysteries" of a woman's body--puberty, menstruation, pregnancy, lactation, and menopause--are seen as times of power and growth. Menopause is an opportunity for conscious change, not a disease to be treated. PMID- 10380446 TI - Homeopathy. A theoretical framework and clinical application. AB - The use of homeopathic remedies for the treatment of mastitis is described. The basic principles of homeopathy are discussed, including the simillimum, the minimum dose, the single remedy, the whole person, the vital force, susceptibility, and constitutional treatment. Homeopathic research trials and papers are examined and discussed. The author explains how homeopathy can be incorporated into midwifery practice and applies this to the treatment of mastitis. Specific indications in the application of 19 homeopathic remedies for mastitis, breast abscess, and lactation difficulties (including problems with supply and painful nipples) are cited. Keynote symptoms are presented in an easy access repertory. General guidelines for potency and dosage protocol are given. PMID- 10380447 TI - Homeopathic remedies in prenatal care. AB - The basic concepts of homeopathy are presented, including the vital force, the Law of Similars, the Law of Proving, and the Law of Potentization. The method by which the practitioner applies these laws in a clinical setting in order to choose a homeopathic remedy is described. Careful history taking and observation of the client to ascertain the etiology and location of a complaint, associated sensations, factors that aggravate or ameliorate symptoms, the emotional and mental state, general observations, and strange, rare, and peculiar symptoms are stressed. Specific remedy recommendations for the treatment of leg cramps and other pregnancy-induced discomforts, such as anemia, herpes, nausea and hyperemesis, ptyalism, and pica are included. The use of remedies to turn breech and other malpositioned babies prior to term is presented, as well as discussions on the induction of labor and homeopathic intervention for premature labors. A description of how remedies are administered, handled, and stored is included. Finally, qualifications to practice homeopathy and legal issues for midwives are discussed. PMID- 10380448 TI - Therapeutic touch. A viable link to midwifery practice. AB - Through the use of their hands, their intelligence, and their compassion, midwives have traditionally taken a noninterventionist approach to labor and childbirth. Therapeutic Tough, known by its practitioners as simply TT, is a contemporary healing modality through which a practitioner's hands are used to facilitate healing. Midwives and practitioners of TT share an appreciation of the value of a holistic and noninterventionist approach to maintaining patient health. This article explores the potential use of TT as a tool to complement the practice of midwifery. It provides an explanation of the theories underlying TT, the history of its development, and the process involved in its practice. Additionally, the results of studies both supportive and critical of the practice of TT are analyzed. Finally, possible applications of TT to the practice of midwifery are suggested. PMID- 10380449 TI - Alternative healing in nurse-midwifery practice. AB - This Clinical Practice Exchange focuses on alternative healing in nurse-midwifery practice. It features interviews with six certified nurse-midwives (CNMs) who practice complementary therapies (CTs). The healing modalities they use include homeopathy, Healing Touch, hypnosis, herbal healing, mindfulness meditation, and water healing. The CNMs discuss their training to practice CTs, how they use alternative healing with clients, and how they integrate this with midwifery practice. The interviews are followed by an Alternative Healing Directory composed of 37 CNMs who responded to a Call, which appeared several times in Quickening and JNM. Each midwife's listing includes contact information, CTs practices, and special interests in networking with other CNMs about alternative healing. The JNM hopes that this Directory will be a catalyst for networking and communication that will move forward the discussion, practice, and research of alternative healing within the midwifery community. PMID- 10380451 TI - The use of complementary therapies in midwifery in the UK. AB - Midwives in the United Kingdom (UK) are autonomous, independent practitioners and the lead professionals in normal pregnancy and childbirth. Changing Childbirth, a government report, gave a recommendation that women should have continuity of care. Midwives have recognized the ability to implement complementary therapies in health care and have succeeded in forming the Complementary Therapies in Maternity Care National Forum (May 1988). The National Health Service Confederation identified midwives as the highest users of complementary therapies in the health care services. Midwives are in a position to incorporate complementary therapies into their practice in conjunction with the rules and guidelines promulgated by the UK Central Council for Nursing, Midwifery, and Health Visiting. Highlighting the Complementary Therapies in Maternity Care National Forum underscores the increased use of therapies by midwives in the UK. Documentation of complementary therapies used in midwifery practice has resulted in some evidence-based practice for reference. Caseload midwifery (the progressive approach of smaller teams of midwives, who are community-based) and education can play key roles in integrating complementary therapies into midwifery, providing women with more choice, and achieving greater client satisfaction from the childbirth experience. Success is also dependent on government commitment and involvement. PMID- 10380450 TI - Oral evening primrose oil: its effect on length of pregnancy and selected intrapartum outcomes in low-risk nulliparous women. AB - Evening primrose oil is widely used by many midwives to hasten cervical ripening in an effort to shorten labor and decrease the incidence of postdates pregnancies. Although its efficacy has been studied in the relief of symptoms of a number of medical conditions, its use has not been well studied, if at all, for the purpose of cervical ripening. The purpose of this study was to investigate the effect of oral evening primrose oil on the length of pregnancy and selected intrapartum outcomes in low-risk nulliparous women. A two group retrospective quasi-experimental design conducted on a sample of women who received care in a birth center, compared selected outcomes of 54 women taking evening primrose oil in their pregnancy with a control group of 54 women who did not. Findings suggest that the oral administration of evening primrose oil from the 37th gestational week until birth does not shorten gestation or decrease the overall length of labor. Further, the use of orally administered evening primrose oil may be associated with an increase in the incidence of prolonged rupture of membranes, oxytocin augmentation, arrest of descent, and vacuum extraction. PMID- 10380452 TI - [Eosinophilic myalgia syndrome]. PMID- 10380453 TI - [AIDS and tuberculosis in the light of spreading drug abuse]. PMID- 10380454 TI - [Non-narcotic analgesics and non-steroid anti-inflammatory drugs and gastric erosions and ulcer]. PMID- 10380455 TI - [Clinical aspects of somatic disease and internal course of the disease (heart ischemia and bronchial asthma model)]. AB - Criteria of subjective severity (SS) and subjective control of the condition (SC) associated with establishment of the disease internal picture (DIP) were determined in 67 patients with verified coronary heart disease (CHD) and 68 with verified bronchial asthma (BA). Essential for SS were: severity of the subjective symptoms, features of the somatic condition debut, the speed of the disease progression. Essential for SC were the following components: effectiveness of self-care, severity of noticeable or ugly symptoms, features of the triggers (attack provokers). SS and SC criteria can be used for differentiation of the interventions to change the patient's attitude to their disease and therapy in the direction of more compliance with the doctor. PMID- 10380456 TI - [Cardiological aspects of adaptation to continuous low-energy nutrition]. AB - Monitoring of some functional, somatometric and ergometric parameters has been performed in 16 healthy volunteers kept on low-calorie diet (energetic deficit 2000 kcal/day) for 49 days. The obtained specific staging of adaptation, morbid and recovery reactions of the circulation can serve the basis for design of valid approaches to reducing and rehabilitation diet therapy. PMID- 10380457 TI - [On the pathogenesis and treatment of erosive gastritis and duodenitis]. AB - The examination of 52 patients with erosive lesion of the gastroduodenal zone (39 patients with erosive gastritis and 13 with erosive duodenitis) has found association of gastroduodenal erosions with changes in intragastric and intraduodenal pressure, gastric concentrations of biliary acids, hormone secretion (gastrin, insulin, hydrocortisone, thyroxine, thyrotropine), with low functional capacity of the mucus in gastric contents, the presence of Helicobacter pylori. Combined treatment with polyphepan and cerucal produced good response in erosive gastritis and duodenitis. Along with relief of the clinical symptoms and erosion epithelization in 96% of the patients, there was a reduction in the cavity pressure and bile acids concentration in gastric contents, normalization of pyloric function, improvement of the function of gastric mucosa protective barrier. PMID- 10380458 TI - [The interconnection between the severity of meningococcal infection and the endotoxicity levels and patient's blood complement]. AB - Seventy-eight patients with severe systemic meningococcal disease admitted to the Intensive Care Unit of the Second Moscow Hospital for Infectious Diseases were divided into four groups by complications of their disease: patients with refractory septic shock (RSS)--group 1; patients with early septic shock (ESS)- group 2; patients without shock but with severe mental disorders--group 3; patients without any of these complications--group 4. The LPS concentration in plasma was assessed by chromogenic method. Initial LPS levels in plasma of group 3 or group 4 patients (170 ng/l, median value and 360 ng/l, respectively) were greater than those of healthy donors (LPS < 15 ng/l). LPS concentration was significantly greater in group 2 (920 ng/l) or group 1 (12,400 ng/l). LPS levels declined exponentially in all the patients. The half-life was calculated to be 1.4 (+) -0.3 h. In group 2 and 1, respectively, the classical pathway complement activity in patients' serum was 50 and 10% of normal control values. To estimate significant prognostic factors for fatality in our patients, specificity and factor fatality difference of various clinical and laboratory factors were calculated. The cut-off LPS value for development of ESS was 600 ng/l and that for development of RSS and death was 8000 ng/l. For the prediction of fatality using the former cut-off value of LPS, sensitivity was 84% and specificity 100%. Using plasma complement activity (cut-off--15% of normal value) for prediction, sensitivity was 75% and specificity was 100%. Other factors (platelet and WBC count, blood pH, BP, etc.) had lower predictive power. Thus to date plasma endotoxin level and complement activity are the best prognostic factors in meningococcal disease. PMID- 10380459 TI - [The mechanism of normolipidemia among the Northern area ethnic groups]. AB - Biochemical composition of the bile and blood serum was investigated in Evenkia and Yakutia residents and immigrants. Blood lipids in the immigrants were much higher while bile lipids lower than in the natives. It is suggested that normolipidemia in the residents may be determined by intensive synthesis of biliary acids and other lipids in the liver and their secretion into the bile. Later, high content of primary cholates (17%) was found in the bile of residents which was not confirmed in the immigrants. PMID- 10380461 TI - [Clinical and prognostic significance of the degree of delayed ventricular function and vegetative regulation of cardiac rhythm in myocardial infarction patients]. AB - Ventricular tachyarrhythmias (VT) are responsible for many sudden deaths in postmyocardial infarction (PMI) patients. The aim of the study was to investigate clinical and prognostic significance for such patients of delayed ventricular activity and vegetative regulation of cardiac rhythm. Signal-averaged ECG, mathematical analysis of cardiac rhythm with measurement of variation range, mode, index of regulatory systems tension, 24-h ECG monitoring were made in 90 patients with acute large-focal myocardial infarction (MI) on the disease day 1, 15, 40 and 365. Signal-averaged ECG registered on MI day 15 was found most informative for identifying the risk to develop VT. Combined application of diagnostic techniques brings more information than each of them alone. PMID- 10380460 TI - [The efficacy of flonivin BS in the treatment of intestinal dysbacteriosis]. AB - The paper presents results of intestinal dysbacteriosis (ID) treatment with flonivin in 25 patients. The diagnosis was made clinically and bacteriologically (fecal examinations). Flonivin BS proved to be an effective biological preparation for management of ID stage I and II. It is composed of bacteria which do not conflict with macroorganism, contribute to normalisation of intestinal biocenosis, is especially beneficial in ID induced by long-term administration of antibiotics. PMID- 10380462 TI - [Addison's disease of tuberculous etiology erroneously diagnosed as secondary porphyria cutanea]. PMID- 10380464 TI - [Research and practical conference on "The Modern Technologies in Rehabilitation Medicine]. PMID- 10380463 TI - [The concept of the relationship between the human body and Helicobacter pylori]. PMID- 10380465 TI - [Old slavonic medicine]. PMID- 10380466 TI - [Memorable dates in the history of medicine in 1999]. PMID- 10380467 TI - [Stump curettage cytologic examination of excised tumor is more sensitive than histologic examination]. AB - Some malignant lung tumors relapse on the cutting line in spite of a negative histologic stump. Histologic examination is limited to only few sites of excised sample. Conversely, stump curettage cytologic examination is useful to examine the whole area. We conducted a simultaneous histological and cytological study to assess the value of stump curettage cytologic examination. Forty-two staple excised peripheral small lesions (< 3 cm in diameter) from 35 patients, aged 50 to 82 years, were assessed. Ten were subject to VATS and 32 to thoracotomy. Preoperative diagnoses were: 15 undiagnosed lesions (lung cancer: 8, benign lesion: 7), 18 lung cancers and 9 metastatic cancer. The whole area of the excised stump was curetted before observing the cross section to prevent malignant cell contamination. After that, the sample was cut vertically to staple line and pathologically examined. Thirty-eight percent (11/29) of histologically negative stumps were cytologically positive. Twenty-seven malignant lesions underwent only excision and 6 stumps were finally cytologically positive. Two of 6 cytological positive lesions relapsed at the staple site were excised at the second operation. Stump curettage cytological examination was more sensitive than histological examination to detect malignant cell contamination. PMID- 10380468 TI - [Open heart surgery without the use of homologous blood in infants and young children: effectiveness of the intraoperative autologous blood donation from arterial monitoring line]. AB - Open heart surgery without the use of homologous blood transfusion was attempted in 81 pediatric patients weighing 5.5-14.9 kg. Autologous blood was donated from arterial monitoring line after induction of anesthesia. This donated blood, actual volume of 9.2 +/- 1.7 ml/kg, was reinfused following the cessation of extracorporeal circulation. Hemodilution resulting from autologous blood donation was well tolerated by all patients. Sixty-eight patients (84%) were operated on successfully without the need for homologous blood. These data suggest that autologous blood donation from arterial monitoring line is a safe and effective method to avoid homologous blood transfusion especially in infants and young children undergoing open heart surgery. PMID- 10380469 TI - [Two cases of thoracic aneurysm with aberrant origin of the aortic branches: diagnosis and strategy]. AB - We present two cases of thoracic aortic aneurysms with anomalous origin of the aortic arch branches. One was a 72-year-old female with a ruptured descending thoracic aneurysm and aberrant origin of the right subclavian artery. The other was a 64-year-old male with a saccular distal arch aneurysm and aberrant origin of the left vertebral artery. Preoperative examinations included angiography, computed tomography (CT), three dimensional enhanced CT (3DCT), digital subtraction angiography (DSA), and magnetic resonance imaging (MRI). Understanding the structure of neck vessels is important in deciding where to clamp or to reconstruct in surgical repair of the aortic arch. 3DCT was the most useful examination for this understanding. PMID- 10380470 TI - [Application of modified ultrafiltration to cardiac surgery in adults]. AB - Modified Ultrafiltration (MUF) was developed for blood concentration and reduction of postoperative edema in cardiac surgery in children. Its beneficial effects on postoperative hemodynamics have been reported. We applied MUF to cardiac surgery in adults and evaluated its usefulness. Between August, 1995 and April, 1997, MUF was performed in 41 adult patients. MUF was carried out immediately after the cessation of cardiopulmonary bypass. The mean fluid volume removed was 1,135.9 +/- 274.1 ml. The patient's haematocrit significantly increased from 23.2 +/- 2.6% to 26.9 +/- 3.2% (p < 0.0001). The dose of inotropes administered was maintained constant during MUF, and no changes were observed in CVP and the heart rate. However, the systolic blood pressure increased from 99.5 +/- 14.7 to 113.2 +/- 16.2 mmHg (p < 0.0001) and cardiac index from 4.2 +/- 0.9 to 4.9 +/- 1.3 l/min/m2 (p = 0.0006). It was suggested that MUF was an useful technique of haemoconcentration and appeared to have beneficial effects on postoperative hemodynamics in adult cardiac surgery. PMID- 10380471 TI - [A case of coronary artery bypass grafting in a patient who has a rosary-like coronary lesion with Buerger's disease]. AB - We report a case of coronary artery bypass grafting using left intra thoracic artery (LITA) in a 63-year-old male with Buerger's disease. His coronary angiography showed a long lesion seemed rosary in the left anterior descending (LAD). At operation, the surface of the coronary artery was dark red with white speckles. The LAD was opened distal end of disease, the intima of the coronary artery made much soft thin fold. The disease was never atheromatous. His coronary angiography showed a very rarely finding that seemed rosary. PMID- 10380472 TI - [A case of surgical resection of metastatic multiple lung pheochromocytoma with long-term survival]. AB - A case of resection of metastatic multiple lung malignant pheochromocytoma is presented. Seven years before, total resection of the right adrenal gland and the right kidney was performed to remove a pheochromocytoma. During the post-surgical observation in the urology outpatient clinic, the blood noradrenalin concentration increased and multiple lung metastatic nidi were observed by chest X-ray. An excision was made to remove 3 wedge-shape metastatic nidi from both lungs. In the third year after the removal of the pulmonary metastatic nidi, a resection was again performed due to recurrence in the left retroperitoneal lymph node. Another recurrence was found in the S6 liver 1 year after the removal of the lymph node, and TAE and PEIT were conducted. When the recurrent metastatic nidi were found, the blood noradrenalin concentration was elevated, and it normalized after treatment. In this case, careful and long-term observation including monitoring the blood noradrenalin concentration in the outpatient clinic and positive surgical treatment proved effective. The patient is currently in good health without signs of recurrence 12 years after the removal of the primary adrenal nidus and 6 years after the removal of the pulmonary metastatic nidus. PMID- 10380473 TI - [Axillary artery perfusion for the extensive arterial vascular disease]. AB - Usefulness of axillar artery perfusion for the cases with severe systematic atherosclerosis was reported. It is generally accepted that the femoral artery is a common arterial cannulation site when performing the surgery of ascending aorta and total aortic arch. However, atheroembolism is one of the most severe complication for the patients with extensive arterial vascular disease by using femoral arterial perfusion. Axillar artery perfusion can prevent these complications, and the perfusion through the artificial graft anastomosed to the axillar artery can also avoid the malperfusion of the vertebral artery and axillar artery. We concluded that the axillar artery perfusion via artificial graft is useful alternative for aortic surgery. PMID- 10380474 TI - [Histidine buffered cardioplegic solution (HBS) provides effective myocardial preservation with wider safety margin in patients with prolonged ischemia]. AB - Increasingly complex techniques of cardiac surgery often require prolonged myocardial ischemia. We previously reported a better myocardial preservation with histidine containing cardioplegia (HBS) in human open heart surgery. To see a safety margin of this cardioplegia for prolonged myocardial preservation, this study was retrospectively done. One hundred twenty-six patients received either intermittent multidose (20-30 minute) cold blood cardioplegia (CBC) plus topical cooling (aotric cross clamp time (ACC) < 120 minutes, n = 63) or HBS (n = 63). HBS group was divided into two groups with either short ACC (< or = 120 minutes, HBS-S, n = 46) or long ACC (> 120 minutes, HBS-L, n = 17). Cardiac index (C.I.) and dopamin/dobutamine requirement were measured 3, 6, and 12 hours post-bypass. Incidence of homologous blood transfusion was also studied. There was two deaths due to LOS in HBS-S group; four patients in HBS group required 5 DC cardioversion, whereas six patients required a total of 12 DC cardioversion in CBC group. Functional recovery were significantly better with significantly lower inotropic requirements in HBS-S group than CBC group and HBS-L group. Although preoperative ejection fraction and C.I. were significantly lower in HBS-L group, post-operative cardiac function and inotropic requirements in HBS-L was comparable to that seen in CBC group. We conclude that the highly buffered histidine crystalloid cardioplegia solution provides effective myocardial preservation with a wider safety margin for prolonged myocardial preservation in open heart surgery. PMID- 10380475 TI - [Experience with repair of muscular trabecular ventricular septal defects]. AB - We reviewed the records of 9 pediatric patients with muscular trabecular ventricular septal defects undergoing repair between April 1994 and June 1998 (mean age 2.6 +/- 3.1 years, mean weight 9.0 +/- 5.2 kg). The prevalence rate for muscular trabecular defects in the patients undergoing open-heart surgery for congenital heart disease was 2.0%. Although only 6 of the 9 patients were diagnosed as having muscular trabecular defects preoperatively, 60 degrees left anterior oblique and 30 cranially tilted projections of left ventricular cineangiocardiogram were useful to detect these defects. The technique of filling the left heart with blood by stopping to vent the left heart and inflating the lungs during the last one or two ligatures in closure of the defects was also useful to detect these defects intraoperatively. In closure of muscular trabecular defects, division of some trabeculations including a moderator band enabled complete repair through a right atriotomy. The nearer the inferior border of the ventricular septal defects were to the heart apex, the more postoperative residual shunts were left. We consider that apical left ventriculotomy may be needed in apical defects, although the right atrial approach is satisfactory for most muscular trabecular defects. PMID- 10380476 TI - [Intrapulmonary hematoma surrounding the stapled line after video-assisted thoracoscopic bullectomy for spontaneous pneumothorax]. AB - A 52-year-old man presented at our hospital with hemoptysis three months after undergoing a video-assisted thoracoscopic bullectomy for spontaneous pneumothorax. A chest X-ray showed a localized infiltrative shadow in the right upper lobe of the lung. The chest CT findings revealed a mass-like lesion surrounding the staple which had been used during the bullectomy. He therefore underwent a pulmonary resection including the lesion due to the continued hemoptysis that did not improve even after treatment by hemostatic agents and bronchial arterial embolization. The resected specimen revealed an intrapulmonary hematoma with severe inflammation. The late onset hematoma which was induced by the stapler may thus be a rare complication in video-assisted thoracoscopic surgery. PMID- 10380477 TI - [Coronary artery bypass grafting for a patient with angina pectoris and ulcerative colitis]. AB - The implementation of coronary artery bypass grafting for angina pectoris with ulcerative colitis has been rarely reported. A 63-year-old man has a past history of acute myocardial infarction in 1984 and ulcerative colitis since 1988. Coronary angiography and cardiac catheterization showed total obstruction of segment 2, 95% stenosis of segment 6, 75% stenosis of segment 7 and total obstruction of segment 12 with LVEF 23%. Coronary artery bypass grafting was performed under IABP support and cardiopulmonary bypass with aprotinin infusion after an inflammatory reaction of ulcerative colitis was adequately suppressed. Ulcerative colitis was controlled by administering 40 mg of predonisolone during perioperative period. PMID- 10380478 TI - [A case report of a coronary fistula from left coronary artery to left ventricle]. AB - The patient is a 44-year-old man. He has had chest pain on effort since 1994, diagnosed as unstable angina in 1996. He was found to have three vessel disease on coronary angiography and was sent to TWMC for operation. CABG was performed for the right and left anterior descending arteries, and TMLR for the posteroinferior region because the left circumflex artery was fine and ungraftable. Both the grafts were patent and there was improved LV function. Interestingly, we found a coronary fistula running from the left coronary artery to the left ventricle. As far as we can tell from our literature searches, this is the first time that a coronary fistula has been found after TMLR. PMID- 10380479 TI - [A case of cor triatriatum with atrial septal defect in an adult]. AB - A surgical treatment of a 54 year-old female with cor triatriatum was reported. Two dimensional echocardiography had demonstrated an abnormal septum in the left atrium. The abnormal septum had a 2 x 1 cm fenestration and existed between the left and right pulmonary veins. Preoperatively, we had misjudged the septum as ASD and misdiagnosed this case as PAPVC. During surgery we found that there was the accessory atrial chamber posterior to the true ASD. The accessory chamber received right pulmonary veins and connected to the left atrium. The left pulmonary veins connected to the left atrium normally behind the abnormal septum. We diagnosed this case as Lucas-Shmidt IIIA1 type cor triatriatum with ASD. The abnormal septum was resected, and the ASD was closed with a bovine pericardium. The postoperative course was uneventful and she was discharged 19 days after the operation. PMID- 10380480 TI - [Breast cancer at the site of an implanted pacemaker]. AB - We report here an 81-old-female patient who had a permanent pacemaker implanted in the right chest and who developed breast cancer near the site of the implanted generator. The cancer was diagnosed as stage I adenocarcinoma and radical mastectomy preserving pectoral muscles was indicated. During temporary pacing via the femoral approach, the pacemaker lead was transferred to the left subclavicular area crossing before the sternum and the generator was reimplanted in the left chest without use of lead extension kit. After reimplantation of the generator, radical mastectomy was performed. Clinical course was uneventful after the operation without infection or pacing failure. For the patient who needs surgical procedure in the site of implanted pacemaker generator, this technique of reimplantation is one of the useful choices. PMID- 10380481 TI - [Reoperative off-pump subclavian-coronary artery bypass grafting in an elderly patient with left internal thoracic artery stenosis]. AB - A 80-year-old Japanese female was diagnosed to have angina pectoris and admitted to our hospital. She had been operated on with mitral valve replacement and coronary artery bypass grafting to right and circumflex coronary artery 4 years before. The coronary angiogram showed significant stenosis with severe calcification in the left anterior descending coronary artery, and it was unsuitable for catheter intervention. The patient also had stenotic left internal thoracic artery and multiple cerebral infarction, but successful off-pump subclavian-coronary artery bypass grafting using saphenous vein graft through small thoracotomy was performed without new neurological deficit. This procedure is useful for patients with left internal thoracic artery unsuitable for MIDCABG, due to quality, size, or injury during preparation. PMID- 10380482 TI - [A case of pure red cell aplasia with hypogammaglobulinemia appearing after thymo thymectomy]. AB - We present a case of 83-year-old woman with pure red cell aplasia appearing eight months after thymo-thymectomy for an invasive thymoma. She underwent thymo thymectomy for an invasive thymoma in July 1996. Preoperative examination revealed neither anemia nor hypogammaglobulinemia. About eight months after the operation, she was readmitted because of anemia and hypogammaglobulinemia. Bone marrow aspiration revealed absence of erythroblasts and chest CT revealed norecurrence of thymoma. Her anemia had responded to ciclosporin. PMID- 10380483 TI - [A case of arteriovenous fistula of the lung]. AB - A 29-year-old male was admitted to our hospital for further evaluation of left sided paresis, cyanosis, clubbing finger. The laboratory data revealed polycythemia and hypoxemia. Cerebralarteriogram showed right middle cerebral artery occulusion. Cardiofunctional test showed atrial fibrillation, lower left ventricular function. Cardiac catheterization, pulmonary arteriography and three dimensions CT were performed. Right to left shunt rate was 20.4%. A single large pulmonary arteriovenous fistura with a feeding artery (A10) and a draining vein (V10) was found clearly. In this cases, arteriovenous fistula was large, blood flow was thought to be rapid. We thought transcatheter embolization was not useful. And we performed right lower lobectomy. Postoperative course was not eventful. Cyanosis disappeared, clubbing finger was cured. PMID- 10380484 TI - [A case of pulmonary arteriovenous fistula associated with cerebellar abscess]. AB - We report a rare case of pulmonary arteriovenous malformation (PAVM) with cerebellar abscess. The patient was 38-year-old woman who admitted to the local hospital for headache and fever. Subsequently, her condition became critical with consciousness disturbance, and hypoxemia. Brain computed tomography (CT) and chest CT revealed cerebellar abscess and PAVM. She was referred to our hospital for the surgery. Pulmonary angiography demonstrated multiple pulmonary arteriovenous fistulas in the right middle lobe and a single nodular pulmonary arteriovenous fistula in the right S8 (10 x 10 mm). After the drainage for the brain abscess, lobectomy of the right middle lobe and the excision of the nodal fistula in the right S8 were successfully performed in the two-staged operation. The patient has done well with no complication and her hypoxemia was improved. PMID- 10380485 TI - [Quality registries are a gold-mine of information that should be better used. Knowledge of improvement is acquired by means of special projects]. PMID- 10380486 TI - [The concept of compulsion in psychiatry: going back to the terminology of LSPV would make the meaning clearer]. PMID- 10380487 TI - [Swedes living in costal regions are healthier and live longer. The Roseto effect in Finland?]. PMID- 10380488 TI - [In otitis: the current principal rule is still valid]. PMID- 10380489 TI - [The "fundamentalists" of breast feeding]. PMID- 10380490 TI - [Are patients with claudication really obese?]. PMID- 10380491 TI - [New materials improve joint prostheses. Metals, polymers, ceramics and composite materials extend the durability]. AB - Advances in our knowledge of multfactorial host-biomaterial interactions have improved the outcome of joint replacement surgery. Fixation and aseptic loosening are the principal foci of research. Wear debris from articulating parts is a major cause of prosthetic loosening. Tribological analysis, and the development of new materials and surface finishes have reduced wear particle production. The use of modular implants increases flexibility and facilitates surgical fit, but also introduces new problems such as dissociation of components, corrosion and wear. In the future, as design, biomaterials and surgical technique become further optimised, arthroplasty may also become an alternative for use in younger patients. PMID- 10380492 TI - [Many years of registration have improved the quality of hip arthroplasty]. AB - The Swedish Hip Replacement Registry has defined the epidemiology of total hip replacement in this country. Most hip replacements are fully cemented. Serious complication and revision rates associated with total hip replacement have declined significantly despite increase in the number of patients at risk. During the past five-year period, only 8(9 per cent of hip replacement procedures have been revisions. Although aseptic loosening with or without osteolysis is the major problem, accounting for 73 per cent of revisions, its incidence has decreased four-fold over the past 15 years to less than three per cent at 10-year follow-up. Quality of the surgical technique is the crucial determinant of the risk of revision due to aseptic loosening, but the choice of implant is also important. Total hip replacement practice in Sweden has improved due to the information available from the registry concerning individual risk factors, implant safety, and the efficacy of improving surgical and cementing techniques. PMID- 10380493 TI - [New system for testing mechanical heart valves. Self-monitoring at home of anti vitamin K therapy means greater freedom for the patient]. AB - Self-monitoring of anti-vitamin K treatment by patients with heart valve prostheses is a good alternative to hospital control. Self-monitoring at home allows patients more freedom and opportunity to take greater responsibility for their treatment. Experience from over a years' complication-free treatment of 12 patients is reported in the article. PMID- 10380494 TI - [Better AVK treatment with self monitoring. Dosage can be regulated in time]. AB - As long-term anticoagulant treatment, with warfarin for instance, is associated with a risk of both thrombotic and thrombolytic complications, blood testing for dose regulation is necessary at 3-8-week intervals, which is expensive and inconvenient for patients who must take time off work and travel to and fro. A new technique, using small portable monitors designed for home use by patients, makes self-management of anticoagulant treatment possible. In Germany, over 25,000 patients had their own monitor by the end of 1998. After appropriate instruction, the German patients are able to monitor their prothrombin time and adjust their anticoagulant treatment accordingly. In case of problems they contact their GP. In a two-year pilot study conducted at the Anticoagulation Clinic of Sahlgrenska University Hospital, Gothenburg, in 1996-98, where 51 patients on long-term anticoagulant treatment were trained in self-management, the results of over 1,000 patient-hours of treatment showed self-management to be at least as safe as management by the clinic. The level of patient satisfaction is high, in terms of safety and freedom from regular hospital attendance during working hours, and the convenience of self-monitoring on holiday or business trips. As the patients do their testing once a week, the risk of complications is also reduced. PMID- 10380495 TI - [Coordinated Swedish transfer is recommended during 1999. Prothrombin complex measurement should be indicated as a quota, not percent]. PMID- 10380496 TI - [A pilot project to learn quality development: care of patients with recently diagnosed rheumatic arthritis]. PMID- 10380497 TI - [Quality projects of the Swedish registry on heart surgery. To study other clinics is to learn, not to judge]. PMID- 10380498 TI - [With the body fluids balanced, vision would come back]. PMID- 10380499 TI - [Nocturnal leg cramp is a common and painful symptom in the elderly. Underlying causes and treatment]. PMID- 10380500 TI - [Quinine/quinidine treatment of cramps of the calf is not supported by the Pharmaceutical Product Agency]. PMID- 10380501 TI - [Midazolam presynaptically inhibits excitatory synaptic transmission in the superficial dorsal horn of the rat spinal cord]. AB - Whole-cell voltage-clamp recordings were made from superficial dorsal horn neurons in thin slices of neonatal rat spinal cord. Spontaneously occurring glutaminergic miniature excitatory postsynaptic currents (mEPSCs) were recorded in the presence of tetrodotoxin, strychnine and bicuculline. Bath-application of midazolam (5-60 microM) reduced the frequency of mEPSCs dose-dependently and this depression was antagonized by flumazenil 15 microM. In 8 neurons studied, midazolam 15 microM affected neither the amplitude profiles nor the mean amplitudes of the mEPSCs, suggesting a presynaptic site of action. In a nominally Ca(2+)-free solution, midazolam still reduced the frequency of mEPSCs, but to a lesser extent than in a standard solution. We conclude that midazolam may presynaptically inhibit excitatory synaptic transmission in the superficial dorsal horn by affecting both Ca2+ entry into the nerve terminals and transmitter release mechanisms downstream to Ca2+ entry. PMID- 10380502 TI - [Intraoperative assessment by laser-Doppler skin blood flowmetry of the efficacy of endoscopic thoracic sympathectomy]. AB - We have investigated whether laser-Doppler (L-D) skin blood flowmetry on the finger could be useful for an intraoperative assessment of the efficacy of endoscopic thoracic sympathectomy (ETS) under general anesthesia. Subjects were 5 young adults receiving ETS for palmar hyperhidrosis. ETS was performed with the patients in the semi-sitting position under one lung ventilation. A pair of LDF probes were placed on the palmar side of the both second fingers. Palmar hyperhidrosis disappeared after ETS in all cases, but compensatory hyperhidrosis developed in the back of the body and the thigh. After completion of ETS on one side, the L-D skin blood flow increased to 267.6 +/- 211.1% on the side of ETS, and it increased in 2 other cases and decreased on the contrary in 3 cases on the other side. After ETS on both sides the L-D skin blood flow increased to 265.0 +/ 185.9% on the side of initial ETS and to 211.4 +/- 172.8% on the side of subsequent ETS. The initial EST induced reflex vasoconstriction on the finger of both sides and also on the toe. Spontaneous fluctuation and reflex vasoconstriction of the skin blood flow were still observed, although the periodicity of spontaneous fluctuation between the right and the left finger was lost in some of the cases. An increase in L-D skin blood flow on the side of ongoing ETS is useful for intraoperative assessment of ETS. PMID- 10380503 TI - [Judicial judgements on anesthesia malpractice in Japan]. AB - We reviewed 75 judicial precedents on anesthetic malpractice during surgical procedure which had appeared in legal journals in the period between 1963 and 1997. Anesthetic techniques employed were: general anesthesia (35 cases), spinal anesthesia (19 cases), local anesthesia (12 cases), and others (9 cases). Anesthesiologists were involved in 16 lawsuits, of which anesthesiologists lost 6 suits between 1986 and 1995. There were 8 cases classified as to be caused by respiratory problems including 2 cases of wrong gas supply. The defendants lost all the 8 cases. On the other hand, the plaintiff lost all the cases of malignant hyperthermia (n = 7). There is a tendency of increase in law suit with general anesthesia. Recent judgments suggested the importance of anesthetic managements, correct recording and appropriate monitoring by anesthesiologist during and immediately after surgery. Spinal anesthesia should be performed by anesthesiologist, and the frequency of anesthetic accident should be decreased. Japan is still in short of anesthesiologists and efforts should be paid to increase the number of anesthesia specialists. PMID- 10380504 TI - [Low flow anesthesia at a fresh gas flow of 10 ml.kg-1.min-1 for hours using time cycled ventilator]. AB - Low flow anesthesia (LFA) at a fresh gas flow (FGF) level of 10 ml.kg-1.min-1 with oxygen flow set at 0.5 ml.kg-1.min-1: 0.5 ml.kg-1.min-1 nitrous oxide and 3% isoflurane was performed using time-cycled ventilator on 10 patients of ASA class I or II, with age of 55 +/- 13 (mean +/- SD) years and body weight of 55 +/- 10 kg for 5 h. Excessive anesthetic gases from the anesthesia gas monitor were led to an expiratory breathing tube. After rapid induction and tracheal intubation, denitrogenation was performed for about 5 min using a 100% oxygen flow of 6 l.min 1 before LFA. The inspired/expired oxygen concentration decreased gradually from 96 +/- 2%/90 +/- 2% at beginning of LFA to 42 +/- 3%/37 +/- 4% at 5 h. The operation was started after 29 +/- 10 min of beginning of LFA. The nitrous oxide concentration reached 37 +/- 4%/35 +/- 4% at the beginning of operation and further increased to 55 +/- 3%/53 +/- 3% at 5 h. The isoflurane concentration reached 1.0 +/- 0.1%/0.8 +/- 0.1% at the beginning of operation and further increased to 1.2 +/- 0.1%/1.0 +/- 0.1% at 5 h. The anesthetic potency was 1.2 +/- 0.1 MAC/1.0 +/- 0.2 MAC at the beginning of operation. The isoflurane vaporizer setting was changed only once in two cases from 3% to 2% exceeding 1.5% in inspired concentration. There was no need to change the flow of oxygen and nitrous oxide for 5 hrs. No SpO2 lower than 95% was observed during this study. This method is a clinically safe, easily applicable anesthesia method and used the smallest FGF reported in LFA without occurrence of low FIO2. PMID- 10380505 TI - [Epidural abscess associated with epidural block in a patient with immunosuppressive disease]. AB - A patient with myeloproliferative disorders and diabetes mellitus received epidural block twice for treatment of the low back and leg pain. The drugs used were 1% mepivacaine 4 ml for the first and 1% mepivacaine 6 ml and dexamethazone 4 mg for the second on the next day. Epidural abscess was noticed 2 days later when pus was aspirated through a block needle. MRI revealed the abscess localized at L5/S1. Intensive treatment including epidural drainage and antibiotics succeeded in healing the abscess. Use of epidural block for immunocompromized patients should be decided carefully. PMID- 10380506 TI - [Anesthetic management of a patient with Saber-sheath trachea]. AB - A 65-year-old man was scheduled for total gastrectomy. Preoperative chest radiograph showed significant narrowing of the trachea. On chest CT scan the trachea was U-shaped (tracheal index = 36%) and was diagnosed as saber-sheath trachea. During general anesthesia we took care to reduce the irritation by the endotracheal tube, particularly during intubation, and to avoid excessively high airway pressure. The trachea was watched carefully by bronchoscopy after intubation and during extubation not to neglect any complication. There was no complication after the operation. PMID- 10380507 TI - [Anesthetic management of patients with malignant pleural effusion undergoing hyperthermic perfusion under thoracoscopy]. AB - Nine patients with malignant pleural effusion due to lung cancer had been scheduled for hyperthermic treatment with warmed distilled water (40 degrees C) under thoracoscopy. This treatment aims to produce adhesion of the lungs to reduce pleural effusion. To evaluate the risk of general anesthesia for patients with lung cancer at the end stage, we examined the problems of perioperative management. Seven out of nine patients were classified into ASA physical status > or = III and seven patients into Hugh Jones > or = III Shapiro's score was > or = 5 in four patients. The average %VC was 60 +/- 16 and % FEV1.0 was 41 +/- 18% (means +/- SE). A double lumen endotracheal tube was inserted and anesthesia was maintained with inhalational anesthetics. In two cases, one-lung ventilation could not be maintained because of severe hypoxemia during hyperthermic perfusion. Hypertension occurred in three cases and hypotension in one by direct heat stimulation of the cardiopulmonary system. Although their preoperative risk was poor, there were no major complications and the quality of life was improved. We stress that careful anesthetic management is important for avoiding hypoxemia and hemodynamic instability during this treatment. PMID- 10380508 TI - [Two cases of hearing disorder following general anesthesia]. AB - Hearing impairment is not often considered as a potential complication of general anesthesia, despite several reports of post-operative sensorineural hearing loss. These disorders have occurred after otological as well as cardiobypass surgery. We experienced two patients both of whom had undergone orthopedic surgery. In both cases the patients experienced bilateral reversible hearing impairment after general anesthesia with nitrous oxide. It is likely that a change in the middle ear pressure as a result of Eustachian tube dysfunction may have caused transient conductive hearing loss added to sensorineural hearing disorder. After these cases we interviewed a series of 115 patients who had undergone general anesthesia to assess the extent of this problem. Contrary to our expectation, 7 patients complained of ear fullness or autophony after inhalation of nitrous oxide, although these symptons diminished within 24 hours. It is important to be aware of the possibility of hearing impairment when nitrous oxide is used especially if the patient has a history of a previous middle ear disease. PMID- 10380509 TI - [A case of total intravenous anesthesia with propofol, fentanyl and ketamine for lateral segmentectomy of the liver under pringle maneuver]. AB - We successfully anesthetized an 80-year-old female for Pringle maneuver which was applied at the time of liver transection and consisted of cross-clamping the hepatoduodenal ligament for 25 minutes and releasing the clamp for 2 minutes until the completion of the liver transection. Anesthesia chosen was total intravenous anesthesia with propofol, fentanyl and ketamine (PFK) in combination with epidural anesthesia. The plasma concentrations of propofol at pre-, intra-, and post-Pringle maneuver under the intravenous infusion of propofol at 3 mg.kg 1.h-1 were 0.84, 1.49, and 1.29 micrograms.ml-1, respectively. All operative procedures were done uneventfully and recovery from anesthesia was prompt after the end of propofol infusion. Transient increases in liver enzymes were seen during early postoperative period, but no signs of hepatic failure were observed. In this patient, PFK anesthesia was useful and safe for the liver transection with Pringle maneuver. PMID- 10380510 TI - [Pre- and postoperative complications of elderly patients with femoral neck fractures--a report of 525 cases]. AB - Pre- and postoperative complications of 525 cases of femoral neck fractures in elderly patients were studied. The mean age was 81.5 years. Preoperative complications were found in 94.5% of the patients. Circulatory and respiratory complications were 68.4% and 29.7%, respectively. Dementia was present in 55.6% of the patients. Operations for fractures were performed under spinal anesthesia only or under both spinal and epidural anesthesia. We tried to avoid hypotension and hypoxia during operations and postoperative periods under pulse oximeteric monitoring. In the postoperative period, circulatory and respiratory diseases exacerbated or newly developed in 4.4% and 5.7% of all cases, respectively. Postoperative mortality within a month was 1.0%, and it was 3.6% within a year. Many elderly patients with femoral neck fractures had preoperative complications, but postoperative complications could be avoided by careful management during pre and postoperative periods. PMID- 10380511 TI - [Evaluation of the ketamine maintenance dose in anesthesia using propofol, ketamine and nitrous oxide]. AB - We studied the influence of the ketamine maintenance dose on propofol infusion speed, blood pressure change and recovery time in anesthesia using propofol, ketamine and nitrous oxide. Anesthesia was maintained with ketamine 0.6 or 0.2 mg.kg-1.hr-1. The ketamine maintenance dose exerted no influence on propofol infusion speed, the occurrence of hypertension or hypotension. The recovery time correlated with the total amount of ketamine. From these results we conclude that 0.2 mg.kg-1.hr-1 is an appropriate maintenance dose of ketamine in anesthesia using propofol, ketamine and nitrous oxide. PMID- 10380512 TI - [Ventricular fibrillation during mild hypothermic therapy in a patient undergoing neurosurgery]. AB - A 35 year old male for mild hypothermic therapy for neurosurgery, developed ventricular fibrillation. Total resection of cerebral arteriovenous malformation was performed under mild hypothermic therapy, with the target core temperature of 33 degrees C. Intraoperatively cooling rate was low and a reduction of peripheral temperature associated with metabolic acidosis was noted. After the conclusion of the operation and during transferring the patient to an intensive care unit, multiple ventricular premature beats were noted. Thereafter, ventricular tachycardia developed proceeding ventricular fibrillation. We suggest that careful management of thermal and cardiovascular systems is required to prevent adverse events during mild hypothermic therapy. PMID- 10380513 TI - [Leak test for the internal circuit of anesthesia machines]. AB - In spite of detailed periodic inspection performed by specialized engineers, a great number of anesthesia machines fail to meet the standard of low flow leak test because of leak in the internal circuit. To find out the background and the solution to the problem, we sent questionnaire to 11 major manufacturers and/or dealers each responsible for periodic inspection of anesthetic machines. According to the responses to the questionnaire, the manufacturers and/or the dealers had various methods of internal circuit leak test without a unified standard in detail. The mismatch of the test methods with those anesthesia machines equipped with check valve mechanism has also led to poor evaluation of internal circuit leak. Leak test must be standardized for its appropriate application to work effectively for the problem of leak in the internal circuit of anesthesia machines. PMID- 10380514 TI - [Economical benefit of continuous total intravenous anesthesia]. AB - Total intravenous anesthesia (TIVA) has been recommended in view of avoiding air pollution. However, intermittent administration of anesthetic agents has a large disadvantage of delayed emergence. We reported that continuous TIVA with propofol, ketamine, vecuronium and buprenorphine (PKBp) could bring rapid emergence. In this study, we calculated and compared the cost of anesthesia in the subjects who had undergone general anesthesia either with continuous PKBp or nitrous oxide-oxygen-sevoflurane. In group PKBp subjects, after induction with propofol, ketamine, vecuronium and buprenorphine, anesthesia was maintained with continuous intravenous administration of propofol corresponding to the patient's age using twice step down method; ketamine (240 micrograms.kg-1.h-1), vecuronium (80 micrograms.kg-1.h-1) and buprenorphine (0.4 microgram.kg-1.h-1). Group GOS subjects, after the same induction method, received nitrous oxide, sevoflurane and vecuronium. Moreover, the group GOS subjects were divided to two groups; the high flow GOS (N2O:O2:sevoflurane = 4 l:2 l:30 ml) and the low flow GOS (N2O:O2:sevoflurane = 2 l:1 l:15 ml). Continuous PKBp group showed lower cost than the high flow GOS group. The PKBp group showed lower cost than the low flow GOS group except in patients weighing more than 100 kg. Furthermore, we calculated the cost of continuous PKBp anesthesia in Japan, U.S.A. and U.K. The U.S.A. cost of PKBp was higher than the Japanese and the U.K., because the cost of ketamine in U.S.A. is higher than in the other countries. Continuous PKBp is more economical than the high flow GOS, and continuous PKBp in Japan is more economical than in U.S.A. PMID- 10380515 TI - [The current state of leak in anesthetic machines detected by low flow leak tests]. AB - To assess the current state of leak in anesthetic machines, we selected 66 units of anesthetic machines for inspection and repair from various medical institutions. Based on a newly designed inspection flow chart a low flow leak test for internal circuits of the anesthetic machines was performed. The conventional low flow leak test was also performed for smooth detection of leak for rational evaluation. Only 39% of the anesthetic machines met the standard of the low flow leak tests, and leak was detected in the remaining 61%. The average residual leak mounted to 0.97 l.min-1, with the maximum of 5.3 l.min-1. Canisters, corrugated tubes, and vaporizers were considered the primary causes of leak. After the inspection and repair, leak in 77.5% of the anesthetic machines either disappeared or decreased and the average residual leak dropped to 0.34 l.min-1. However, 47% of the anesthetic machines still failed to meet the standard of the low flow leak tests. To further improve the situation, more detailed inspection and repair are necessary especially for precise detection of the cause of leak in the internal circuit of anesthetic machines which often remains undetected. PMID- 10380516 TI - [Video laparoscopy in abdominal emergencies]. AB - BACKGROUND: Personal experience about the use of video laparoscopy (VL) in abdominal emergencies is reported. METHODS: DESIGN: retrospective evaluation of patients observed in the last years. SETTING: General Surgery I. Policlinico, University of Palermo. SUBJECTS: 61 VL have been performed: 30 acute appendicitis, 21 acute cholecystitis, 4 perforated peptic ulcer, 1 haemoperitoneum by haemorrhaged luteal corpus, 2 pelvic inflammatory disease (PID), 1 terminal ileitis, 1 choledochal perforation after ERCP and 1 bleeding after CVL. INTERVENTIONS: the following interventions have been performed: 22 VL appendectomy, 8 VL-assisted appendectomy, 21 VL cholecystectomy, 1 VL duodenal suture, 3 minilaparotomic duodenal suture, 2 prophylactic VL-assisted appendectomy, in 1 patient with terminal ileitis and in 1 PID, 1 VL partial ovarian resection. In the case with choledochal perforation during ERCP a traditional cholecystectomy was performed with an outer biliary drainage. In the patient with bleeding after CVL the spontaneous haemostasis seen during VL was confirmed by laparotomy performed to exclude baro-haemostasis and to prevent from legal motivation. The procedure was only diagnostic in 1 patient with PID. MAIN OUTCOME MEASURES: the diagnostic and therapeutic value, versus traditional surgery have been valued. RESULTS: VL is useful both for a correct diagnosis and to lead a laparotomy if necessary, allowing an adequate peritoneal exploration and toilet without large incisions; the operation is therefore, in any case, less invasive. CONCLUSIONS: In our experience the usefulness of VL is clear in the suspect of acute appendicitis, acute cholecystitis, perforated peptic ulcer, haemoperitoneum and when diagnosis is not sure and in all other situations in which correct preoperative diagnosis is impossible. So this procedure is useful to make easy a correct diagnosis and to surgical treatment. PMID- 10380517 TI - [The role of echo-endoscopy in the staging of squamous-cell carcinoma of the esophagus. The correlation between the surgical and anatomicopathological findings]. AB - BACKGROUND: Endoscopic ultrasonography (EUS) is a relatively new diagnostic method to assess the extent and the depth of infiltration of esophageal carcinoma. METHODS: From October 1990, 100 patients affected by esophageal squamous cell carcinoma underwent preoperative evaluation with endoscopic ultrasonography, 85 of whom were operated on. The first 23 patients underwent endosonography with an Olympus GF-EUM2 with a 7.5 MHz echo-probe; the remaining 77 patients underwent EUS with an Olympus GF-EUM3 with a 7.5-12 MHz echo-probe. RESULTS: In 33 cases (33%), the procedure was not completed because of the impossibility of passing through the neoplastic stenosis. The depth of infiltration was correctly defined by EUS in 73 of 85 patients (86%) compared with 47% of Computed Tomography (CT) (p < 0.05). Overestimation occurred in 6 patients (7%), whereas underestimation occurred in 6 cases (7%). Lymph-node involvement was correctly classified by EUS in 50 of 57 patients (88%) compared with 39% of CT. CONCLUSIONS: EUS provides a high degree of accuracy in assessing both T and N parameters in staging esophageal cancer. The major problem of the method is still the frequent impossibility of passing through a neoplastic stenosis. PMID- 10380518 TI - [The palliation of dysphagia secondary to esophageal-cardial carcinoma with self expandable metal prostheses. The authors' personal experience with 92 patients]. AB - BACKGROUND: Endoscopic insertion of a stent is an important option in the palliative management of esophageal obstruction and esophagorespiratory fistula. Plastic stents have been available for over 20 years. A new class of self expanding metal stents for palliation of esophageal and cardial cancer is now available. METHODS: Between September 1992 and October 1997, 92 patients underwent implantation of self-expanding metal stents for palliation of dysphagia due to inoperable esophageal or cardial cancer (65 patients) or for locally recurrent carcinoma after surgery (12 patients), laser-therapy (11 patients) or radiotherapy (4 patients). RESULTS: Successful stent implantation was achieved in 89/92 patients (96.7%). After stent implantation the dysphagia score improved from 3.0, on average, to 0.5, on average. Early complications were observed in 4.5% and peroperative mortality was 2.1%. Late complications were observed in 25.6%, with a mortality rate of 1.1%. The mean survival time was 6.9 months. CONCLUSIONS: Self-expanding metal stents are a new effective alternative for palliation of dysphagia due to esophageal and cardial cancers. PMID- 10380519 TI - [The surgery of non-small-cell bronchogenic carcinoma in stage IIIA. An analysis of 150 treated cases]. AB - BACKGROUND AND AIM: The authors report the findings of a retrospective study made of 150 cases of bronchogenic non-small-cell carcinoma at stage IIIA. METHODS: Of the 150 patients treated 130 were male and 20 female. The mean age of the population examined was 55, with a minimum of 28 and maximum of 76. The techniques of exeresis used were pneumonectomy in 70 cases (33.3%) (simple in 50 cases--33.3% and intrapericardial ligation of pulmonary vessels in 20--13.3%), lobectomy in 61 cases (40.6%), lobectomy with associated atypical resection in 9 cases (6%), atypical resection in 6 patients (4%) and bilobectomy in 4 (2.6%). RESULTS: The 5-year survival rate was 16.9%. It was also found that the 5-year survival rate was 20.7% higher for epidermoid carcinoma compared to other histiotypes. The technique used also influenced survival and subjects undergoing pneumonectomy presented a 5-year survival of 29.7% compared to 26.8% for lobectomies associated with atypical resection. CONCLUSION: Surgery of bronchogenic carcinoma at stage IIIA has not obtained promising results in terms of survival. However, no other alternative treatment permits an average 5-year survival rate of 15% to be achieved. PMID- 10380520 TI - [Cervico-mediastinal goiter. Our experience]. AB - BACKGROUND: Personal experience about substernal goiter is reported. Stressing laid on the importance of definition: a goiter that is totally or in the most part below the superior thoracic outlet, with normal vascularization. METHODS: DESIGN: retrospective evaluation of patients observed in the last six years. SETTING: General Surgery I, Policlinico, University of Palermo. SUBJECTS: four hundred ninety-six thyroidectomies have been performed, 32 patients (6.5%) were found to have substernal goiters. The age was between 42 and 86 years (middle age 59). Male/female = 1/1.9. Asymptomatics were 8 (25%). More frequent symptoms were airway compression (34%), hoarseness (9%), pain (9%), thyrotoxicosis (9%) and dysphagia (3%). INTERVENTIONS: total thyroidectomies have been always performed. MAIN OUTCOME MEASURES: the incidence, symptoms, short and long term complication have been valued. RESULTS: There were no postoperative bleeding or lesion of recurrent nerves or definitive hypoparathyroidism. Postoperative hypocalcemia was observed in 9 patients (28%). Only one temporary hypoparathyroidism (two months) was observed. In 2 patients the histologic examination revealed a papillar carcinoma. There were no intraoperative deaths. CONCLUSIONS: In personal experience the presence of substernal goiter is an indication for total thyroidectomy. The reasons for treating substernal goiter surgically are the following: no effective medical treatment is available; respiratory compromise, thyrotoxicosis, dysphagia, or malignancy can develop in long-standing goiters; surgery, in skilled hands, presents minimal morbidity. PMID- 10380521 TI - [A computerized multivariate analysis in the assessment of the prognosis of breast cancer after surgical excision]. AB - BACKGROUND: The prognostic parameters most universally accepted as indicative for the assessment of 5 and 10 year survival in breast cancer are considered. METHODS: The aims of this study were to elucidate the relationships among these parameters and to determine the specific interrelations of these prognostic factors. These parameters (age, grading, tumour size, lymphatic and vascular invasion, necrosis, lymph node status and number of nodes positive) are analysed by univariate and multivariate analysis on a series of 75 patients. RESULTS: The analysis performed confirms prognostic accuracy of age, tumor size, number of nodes positive (p < 0.05, p < 0.001, p < 0.003, respectively) when considered singularly but the impossibility of deriving any benefit from the study of their reciprocal interrelations. CONCLUSIONS: This is due to the importance of each parameter assumes independently from the other and to the reciprocal compensation that occurs when all possible interactions are considered. PMID- 10380522 TI - [The use of stereotaxic cytology in the diagnosis of nonpalpable breast lesions. Our experience]. AB - BACKGROUND AND AIMS: The increasingly frequent use of mammography for the early diagnosis of breast cancer and the consequent identification of mammary lesions at a preclinical stage raises the fundamental problem of the differential diagnosis between non-suspected non-palpable lesions (NPL) which can therefore be monitored over time and suspected NPL or definite carcinoma requiring histological confirmation and surgical biopsy. The diagnostic accuracy of mammography alone is not sufficiently high to differentiate benign lesions from malignant or strongly suspected ones. The use of surgical biopsy in the event of suspected NPL could be significantly reduced by the use of stereotaxic cytology which would improve the diagnostic accuracy of mammography. METHODS: The study refers to 72 suspected NPL undergoing surgical biopsy after having performed stereotaxic cytology on a sample taken with a dedicated mammographic device (Mammotest-TRC). RESULTS: The rate of inadequate samples for correct cytological evaluation was 16.1%. Of the 72 NPL undergoing surgical biopsy, 40 (55.5%) were found to be carcinomas and 32 (44.5%) were benign lesions. The sensitivities of mammography alone and cytology alone in identifying infraclinical breast carcinoma were respectively 0.85 and 0.95. If the results of the two methods were evaluated together, the level of sensitivity was 0.98. CONCLUSIONS: The use of stereotaxic cytology enables a marked improvement to be achieved in the diagnostic accuracy of mammography for the identification of suspected NPL to undergo surgical biopsy, notably reducing the cost of biopsy (number of benign lesions for each carcinoma diagnosed) and consequent discomfort for patients. PMID- 10380523 TI - [An update in the treatment of intra-abdominal abscesses]. AB - BACKGROUND AND AIMS: Intra-abdominal abscesses represent a relatively severe complication in gastroenterological surgery owing to their association with high levels of morbidity and mortality. METHODS: The authors report their experience between January 1990 and January 1996 in 11 patients with intra-abdominal abscesses secondary to emergency surgery for gastroenterology in 10 cases and gynecology in 1 case. After the lesion had been identified using ultrasonography and CT, it was emptied, washed with antibiotic and drained using Seldinger's ultrasonographic and CT-guided technique. Small abscesses (less than 5 mm) were completed removed. RESULTS: The following results were obtained: the immediate disappearance of pain and fever, accompanied by improved general conditions, restoration of canalisation and closure of the abscess cavity (on average between 10 and 15 days). CONCLUSIONS: In conclusion, ultrasonographic-CT guided drainage of postoperative intra-abdominal abscesses, which were previously managed using surgical methods, appears to be the best treatment, relying on the use of imaging techniques and thereby allowing both morbidity and mortality to be reduced. PMID- 10380524 TI - [The usefulness and limits of echo-endoscopy in the clinical staging of esophageal cancer]. AB - Endoscopic ultrasonography (EUS) is a diagnostic method of considerable value for the local staging of esophageal cancer, in particular for T and N evaluation. After an extensive review of the literature, the authors underline the possibility of using EUS to improve the treatment and prognosis of esophageal neoplasms based on the use of various stage-dependent therapeutic strategies. EUS is regarded as a gold-standard technique for esophageal cancer staging in order to select appropriate treatment options, but is currently hampered by the intrinsic difficulty and subjectivity of interpreting ultrasonographic images. In order to ensure safe and reliable data, EUS must be carried out by operators who have undergone suitable training at a specialised centre. PMID- 10380525 TI - [Early gastric cancer. The therapeutic options]. AB - The best therapeutic treatments for early gastric cancer are evaluated. Due to the difficulty of preoperative and intraoperative staging, it's reasonable to consider early gastric cancer as a composite disease; while some mucous localizations can benefit of minor therapeutic treatments, the submucous cancer must be treated as the parietal gastric cancer. PMID- 10380526 TI - [Duplication of the vena cava inferior with a continuation into the vena azygos. A report of a rare case]. AB - The embryogenesis of the inferior vena cava is a complicated process involving development, regression, anastomosis and replacement of three pairs of venous channels (posterior cardinal, subcardinal and supracardinal). A rare case of concurrent duplication and azygos continuation of the inferior vena cava is presented; it is caused by an altered development of subcardinal and supracardinal venous channels. This anomaly, without other congenital malformations (splenic or cardiac), has been previously described only in six cases in the literature. In this case contrast-enhanced CT enabled the correct diagnosis to be made. The subsequent cavography confirmed the CT report. PMID- 10380527 TI - [Recurrent thrombosis of the venous supply of the upper extremity]. AB - The incidence of DVT in axillo-subclavian district is rather low. Up-to-date reports show however it is progressively increasing, owing to the widespread diffusion of intravenous prosthetic devices. While the opportunities of a correct diagnosis are becoming various, particularly because of the development of echo color-Doppler and imaging techniques, there is indecision for the treatment: the question is about a medical and a surgical therapy. Neither the former nor the latter are well established. This case report is about a 61-year-old woman affected by a recurrent DVT of the subclavian and axillary veins, who never underwent operation and/or handling of the venous district. Clinical tests didn't show any well determinate cause of thrombosis. It was decided not to treat the patient surgically, so a medical therapy was undertaken. Now she is completely recovered. PMID- 10380528 TI - [Splenic metastases]. AB - Splenic metastases are found with a frequency varying from 2.4 to 7.1%. The primary tumours most often followed by metastases are breast, lung, pancreas and melanoma. They may also be the direct extension of retroperitoneal tumours and carcinoma of the pancreas. The authors report a case which came to their attention; by examining the literature, they discover the rarity of this pathology which confirms the possibility of this localisation for both intra abdominal and extra-abdominal tumours. PMID- 10380529 TI - [Splenic pseudoaneurysms following acute pancreatitis]. AB - Splenic artery pseudoaneurysms are the most common of visceral artery pseudoaneurysms. Splenic pseudoaneurysms appear to have developed as a consequence of inflammatory processes adjacent to the splenic artery, particularly acute pancreatitis and chronic pancreatitis with associated pseudocysts. They are often asymptomatic and picked up on abdominal examination for ultrasound or CT scanning for other conditions. Complications include rupture with retroperitoneal hemorrhage or intraperitoneal hemorrhage. Two cases of splenic pseudoaneurysms, following acute pancreatitis, are reported between the years 1987 and 1996. PMID- 10380530 TI - [Pheochromocytoma. A case report]. AB - The pheochromocytoma is a catecholamine-secreting tumor, localized in the adrenal gland in 90% of the cases and in extra-adrenal site in the remaining 10%. It can be single or associated with other endocrine neoplasms. On the basis of the case presented, the several clinical manifestations, the treatment of the disease and especially the recent development in imaging as MIBG, TAC, RNM are discussed. PMID- 10380531 TI - The vicissitudes of emergency psychiatry: a service systems perspective. AB - This article, adapted from a speech given at the occasion of the Tenth Anniversary of the American Association for Emergency Psychiatry, highlights the challenges facing this young field. PMID- 10380532 TI - A perspective on voluntary and involuntary outreach services for the homeless mentally ill. AB - Outreach teams use a range of strategies to engage people who are homeless and mentally ill and living on the streets. This chapter describes and evaluates the effectiveness of various voluntary and involuntary approaches and presents a new model program for serving this population. PMID- 10380533 TI - Medical assessment in the psychiatric emergency service. AB - This chapter examines the frequency and consequences of medical morbidity in psychiatric presentations and reviews the factors that should influence policy decisions surrounding "medical clearance" in PES settings. PMID- 10380534 TI - Domestic violence in the psychiatric emergency service. AB - Domestic violence is a frequent precipitant to PES presentation. This chapter discusses case recognition, screening tools, and the debate surrounding mandatory reporting requirements. PMID- 10380535 TI - Psychological trauma and the psychiatric emergency service. AB - Trauma is a frequent precipitant to PES presentation. This chapter reviews the epidemiology, mental health findings, and approach to treatment for a variety of trauma-related syndromes. PMID- 10380536 TI - Disaster mental health: current status and future directions. AB - In times of disaster, emergency mental health services must be delivered in the field. This chapter reviews the current state of the literature and future trends in disaster mental health. PMID- 10380537 TI - An examination of California's civil commitment laws and national future trends. AB - California's civil commitment statute serves as a model for many other states. A move to update the California law may set national precedent. PMID- 10380538 TI - Psychiatric emergency services in the Veterans Health Administration: a review. AB - The VA health care system is the largest source of public mental health care in the country, providing specialty mental health care services to more than 550,000 veterans annually. This chapter reviews the nature and scope of VA psychiatric emergency services. PMID- 10380539 TI - Psychiatric emergency services: policy considerations. AB - Funding for PES occurs through a variety of public and private sources. Potential difficulties associated with providing emergency psychiatric care in a free market economy are discussed. PMID- 10380540 TI - Mobile crisis: moving emergency psychiatry out of the hospital setting. AB - Mobile crisis teams constitute a growing force in emergency psychiatric service provision in the community. The implications of the for-profit, private teams that are joining the long-standing public teams are discussed. PMID- 10380541 TI - [Risk factors in various groups of stroke patients (Analysis of 500 cases of the Budapest Stroke Database)]. AB - Risk factor profile of 500 consecutive acute++ stroke cases in the protocol of the Budapest Stroke Data Bank has been analysed. High frequency of risk factors and additive occurrence have been documented when compared with other stroke registries; hypertension 75%, hypercholesterolemia 68%, ischemic heart disease 61%, hypertriglycerolemia 39%, smoking 38%, serious hypercholesterolemia 36%, diabetes mellitus 30%, peripheral artery disease 10%, elevated hematocrit 7% and elevated number of platelets 7%. More than one risk factors have been registered in 85% of the patients. Three risk factors at the same patient have been found in 28%. The highest number of risk factors is seven at the same patient. The stroke subtypes have been characterized with "cluster-like" association of risk factors. In the hemorrhagic group (9.4% of all cases) hypertension and alcoholism are the main factors; in the atherosclerotic group (49.4%) more male, smoking peripheral artery disease and hypercholesterolemia have been registered; in the lacunar stroke group (27%) high frequency of hypertension, smoking, diabetes mellitus, hypercholesterolemia and hypertriglycerolemia have been found and in the cardiogenic embolia group (10.6%) more female, higher age, ischemic heart disease and atrial fibrillation are the recurrent risk factors. There is no difference between the risk factor profile++ registered in the left versus right hemispheral strokes, in the anterior versus posterior strokes and in the first ever or recurrent strokes. PMID- 10380542 TI - [The pattern of p53 oncoprotein expression in gastric cancer patients]. AB - The p53 gene mutation or p53 oncoprotein overexpression are the most common genetic alterations in human tumors. Its expression is known to be present in a number of tumors however, its prognostic value in gastric cancer is uncertain. The authors studied the pattern of p53 oncoprotein expression in 45 advanced gastric cancer cases. Statistically, the correlation between p53 expression and histologic type of the tumors was significant (p < 0.001): 84% of the intestinal type tumors showed positivity, in half of those (15/31, 48.3%) the positivity involved almost all of the nuclei. This is much higher than any other data in the literature. Moreover, significant correlation was found between tumor stages pT2 3 and p53 oncoprotein expression. There was no significant correlation between lymph node involvement, as one of the most important prognostic factors, and p53 expression. Their results show that the pattern of p53 gene mutation in these gastric cancer patients differs from international data, which might be caused by certain carcinogenic effects: e.g. nutritional customs characteristic of Hungary. PMID- 10380543 TI - [Using the radial artery in myocardial revascularization]. AB - Between March 1996 and April 1998, 10 patients underwent myocardial revascularization using radial artery grafts. The age ranged between 59-81 (mean 67.7) years. The mean left ventricular ejection fraction was found to be 0.53 (range 0.45-0.62). A mean of 3.2 distal anastomoses per patient were performed. There was no operative mortality and no perioperative myocardial infraction was observed. No ischaemia of the hand was noticed. At a mean follow-up of 16.6 months (range 3-27) all patients are alive and free of symptoms. Only one patient had angina at bicycle ergometer test. The precise and atraumatic harvesting of radial artery and use of calcium antagonists prevent vasospasm and early graft occlusion on the short term. To assess the late results longer follow-up is necessary. PMID- 10380544 TI - [Evidence-based medicine in everyday medical practice]. AB - Evidence-based medicine indicates continuous and systematic use of the results of clinical research in everyday clinical practice. It is an important aid to avoid biases of medical decisions caused by following subjective opinion, tradition without criticism or extrapolation from observations at molecular or cellular level. The conscientious use of current best evidence in making decisions about the care of individual patient is an important guarantee of quality. The authors present the details of evidence-based medicine, the main steps of exploring and synthesising evidences. PMID- 10380545 TI - [Critical illness polyneuropathy--report of two cases]. AB - The authors describe the cases of two mechanically ventilated septic patients, in whom developed during their illness with the signs of severe tetraparesis critical illness neuropathy. In both cases the first sign of the neuropathy was the respirator dependency despite the improving neurological status. The severe general condition of the patients and the administered sedatives obscured the neurological signs for a long time, and only the development of a severe tetraparesis raised the suspicion of the critical illness neuropathy. In the first case the diagnosis was made by exclusion, but in the second case it was proved with the help of EMG and the histological examination of the m. biceps brachii. The authors discuss the clinical relevance, neurological and electrophysiological signs and the problems of the differential diagnosis of this disease, which is quite common among the critically ill, septic patients. PMID- 10380546 TI - [Polysomnography--infant mortality]. PMID- 10380547 TI - [Quality assurance in gastroenterology]. PMID- 10380548 TI - [History of mammography in Hungary]. PMID- 10380549 TI - [An update on the antibiotic therapy of tuberculosis]. PMID- 10380550 TI - [New developments and prospects in diabetes mellitus]. PMID- 10380551 TI - [Unruptured cerebral aneurysms. What is the risk of rupture? What is the risk connected with a surgical intervention? A contribution to the international ISUIA study: International Study on Unruptured Intracranial Aneurysms]. PMID- 10380552 TI - [The tolerability and therapeutic efficacy of rifabutin in the treatment of pulmonary tuberculosis]. AB - We treated in our unit 25 patients (15M--10F) affected by pulmonary tuberculosis (TB) with rifabutin (RBT). Chronic liver disease, multidrug-resistant TB and HIV infection were featuring the clinical history of our selected patients. The treatment was carried out using a 150 mg/day dose of RBT, or 300 mg/day in case of MDR (multiple-drug-resistance)-TB or chronic TB. Rifabutin, isoniazid, ethambutol, and a fourth anti-mycobacterium drug were used when treating MDR-TB. Chest X-ray, haematological and bacteriological tests were performed on a monthly basis during the patients' follow up. No side effects were observed; only in two cases, both females, leukopenia occurred, but was not such a reason to modify our treatment plan. As a matter of fact, RBT is well tolerated by patients and it is particularly effective in bacterial eradication. In our experience, RBT did not provide the expected results only in one patient, affected by chronic TB. PMID- 10380553 TI - [Echography and cytologic exam by needle aspirate in the diagnosis of chronic thyroiditis]. AB - PURPOSE: In this study the diagnostic accuracy and clinical, usefulness of sonography and fine needle aspiration (FNA) cytology in the diagnosis of chronic autoimmune thyroiditis (CAT) was evaluated. MATERIAL AND METHODS: An analysis on 220 patients showing elevated serum titers of anti-thyroperoxidase and/or anti thyroglobulin antibodies and laboratory features of hypothyroidism. RESULTS: According to thyroid sonographic character CAT patients were classified into four groups. Group I included patients with an enlarged and diffusely hypoechogenic thyroid gland. Group II comprised patients with a sonographic pattern of multiple focal hypoechogenicities. In group III patients showed a hypoechogenic thyroid gland with single or multiple nodules and scattered hyperechogenic areas. Group IV included patients with a isoechogenic and small thyroid gland characterized by the presence of numerous hyperechogenic spots and strings. DISCUSSION: The most frequent sonographic pattern (72%) associated with CAT was observed in group III patients with a plurinodular and hypoechogenic thyroid. Sonographic patterns observed in group I and II patients are instead highly specific for CAT, since all cases were confirmed at cytology. The high prevalence of cytologic inadequate specimens in group III and IV patients may be related to the higher fibrotic content of their thyroids (multiple hyperechogenicities). CONCLUSIONS: Sonography may represent a useful tool in the diagnostic evaluation of CAT. Information provided by this technique is always to be considered as complementary to that offered by other conventional methods, such as FNA cytology and serum antithyroid antibody measurement. PMID- 10380554 TI - [Tuberculous otitis]. AB - Recent works show an increase of the incidence of extrapulmonary tuberculosis with a peculiar localization to the middle ear, until now considered a rare clinical manifestation. At the Ear, Noise, Throat Clinic of the University of Ferrara a total of 5 cases of tuberculous otitis media were observed during the last 25 years. Aim of the present study is to describe their clinical manifestations, their symptomatic aspects and the way to reach a careful diagnosis and more rapid therapeutic choice. PMID- 10380555 TI - [Alzheimer's disease]. PMID- 10380556 TI - [The therapy of bronchial asthma]. AB - Asthma (Greek word that means "breathlessness" or "open-mouth breath") is a chronic inflammatory disorder of the airways, with extensive infiltration of the airway lumen and wall with eosinophils, mast cells, activated T-lymphocytes. Airway inflammation is associated with airway hyperresponsiveness, recurrent episodes of reversible airflow limitation and respiratory symptoms such as wheezing, chest tightness, breathlessness and cough with mucus production. Curiously, asthma worsens particularly at night and in the early hours of the morning. The current consensus on asthma therapy suggests that pharmacological control of asthma can be achieved with antiinflammatory "controller" medications such as inhaled glucocorticoids and cromones. Short-acting bronchodilators act as "reliever" medications and rapidly reverse acute manifestations of asthma. Asthmatic exacerbations require the repetitive administration of inhaled short acting beta-2-agonist and the early introduction of oral glucocorticoids. Rarely the severity of exacerbation requires the administration of oxygen (that, if available, is not contraindicated), intravenous bronchodilators, glucocorticoids and epinephryne and mechanical ventilation. PMID- 10380557 TI - [Acute purulent meningitis: a clinical and therapeutic update]. AB - A brief overview of the epidemiological and microbiological profile of acute bacterial meningitis during late nineties is outlined, as a basis for an update of the most relevant diagnostic and therapeutic features of this disease, in the different life ages. PMID- 10380559 TI - Removal of Trypanosoma cruzi by white cell-reduction filters: an electronmicroscopic study. AB - White cell (WBC)-reduction filters have been shown to be effective in removing infectious agents from infected blood products. In this study, the mechanisms of Trypanosoma cruzi (T. cruzi) retention by WBC-reduction filters were assessed. Human packed red blood cell (PRBC) and platelet concentrate (PC) samples were contaminated with T. cruzi organisms (Y strain; 3.4 x 10(6)/ml), and then filtered using WBC-reduction experimental filters that provided about 3 log10 WBC removal. Transmission electron microscopy sections showed that T. cruzi parasites were removed from contaminated PRBC and PC samples primarily by mechanical mechanism without interacting with filter fibers or blood cells. In addition, we found that T. cruzi parasites were also removed by a direct fiber adhesion. These data indicate that T. cruzi parasites are removed from infected blood not only by mechanical mechanism but also by biological mechanism probably mediated by parasite surface proteins. PMID- 10380560 TI - Experimental visceral leishmaniasis in high and low antibody-producer mice (selection IV-A). AB - Leishmaniasis is a typical parasite infection whose protective immunity depends on macrophage activation. Susceptibility to Leishmania donovani infection was compared in H (high antibody responder) and L (low antibody responder) mice from selection IV-A. H mice infected intravenously with 10(7) amastigotes of L. donovani were more susceptible to infection than their L counterparts. This higher susceptibility was characterized by a higher splenic and hepatic parasite burden. An increased splenic index was observed in both lines after sixty days of infection. This splenomegaly was caused, at least partially, by an increase in the number of splenic cells as determined by direct counts of cells from spleen. The results show that selection IV-A is susceptible to visceral leishmaniasis, with the H line being more susceptible than the L line. PMID- 10380561 TI - [Human papillomavirus associated with uterine cervix lesions]. AB - It was studied the prevalence of human papillomavirus (HPV) among 228 women with lesions of uterine cervix attending the Ofir Loiola Institute, in Belem, Para, from March 1992 to May 1996. Histopathological examination was performed with all cervical biopsy samples obtained from these patients. In addition, specimens were analysed by both polimerase chain reaction and dot-blot hybridization to detect HPV DNA. The patients were assigned to three groups, according to the diagnosis made by histopathology, as follows: A, including 155 women suffering from invasive epidermoid carcinoma or adenocarcinoma; B, 54 patients having either cervical intraepithelial neoplasia grade II or III; and C, involving 19 women with chronic cervicitis. The prevalence rates of HPV in groups A, B and C were 70.3%, 63% and 36.8% respectively. HPV 16 accounted for 60.4% and 54.5% of types identified in groups A and B, respectively. Altogether HPV types 16, 18 and 33 were detected in 71.4% of positive patients belonging to group C. PMID- 10380562 TI - [Plasmodium falciparum malaria. Quadrennial analysis, during 12 years, of the efficacy of quinine treatment]. AB - The efficacy of quinine for the treatment of falciparum malaria was studied by quadriennal analysis of the medical records of 454 patients admitted to the HDT GO from 1983 to 1994 and treated with identical doses of quinine alone for 7 days. In the quadriennium from 1983 to 1986, 98.4% of the patients became negative by the 5th day of treatment and 8% presented recurrence (R1); from 1987 to 1990, only 72.9% became negative by the 5th day of treatment, 1.4% remained positive until the 7th day (R2) and 9.7% presented recurrence (R1); from 1991 to 1994, 70.1% became negative by the 5th day of treatment, 3.5% remained positive until the 7th day (R2) and 20% presented recurrence (R1). The increase in parasite clearance time with failure up to the 7th day of treatment (R2) and the increase in recurrence (R1) show that P. falciparum is developing resistance to quinine in the region under study. PMID- 10380563 TI - [Phlebotomus (Diptera, Phlebotominae) from Saint Luis Island, Maranhao Gulf region, Brazil]. AB - This study lists 32 species of sand flies, 1 of them belonging to the genus Brumptomyia and 31 to the genus Lutzomyia, distributed among the following subgenera: Psychodopygus (6), Nyssomyia (5), Pressatia (3), Evandromyia (2), Psathyromyia (2), Sciopemyia (2), Lutzomyia (1), Micropygomyia (1), Viannamyia (1), and the groups Oswaldoi (5) and Migonei (3). The sand flies were captured in the wild (forest) and in peridomicile (pigpen, hen house and stable) and intradomicile (bedroom) areas from 06:00 PM to 06:00 AM, once a month, for 4 years on the Island of Sao Luis, Maranhao. All species sampled were present in the forest. Among them, 16 were found in the peridomicile, while 11 were found inside the houses. A total of 22,581 specimens were captured, 65.1% of them in the peridomicile, 17.5% in the forest and 17.4% in the intradomicile. The most common species was Lutzomyia longipalpis (66.4% of the captured specimens), followed by Lutzomyia whitmani (24%) and Lutzomyia evandroi (5.9%). The remaining 29 species represented 3.7% of the total sample. PMID- 10380564 TI - [Usefulness of routine investigation of fungal infections through bronchoscopy in HIV-infected and non-HIV-infected patients in a general hospital, reference to AIDS]. AB - The diagnostic yields and the spectrum of pulmonary fungal-infection obtained in samples collected by fiberoptic bronchoscopy from HIV-positive and HIV-negative patients were evaluated from 1990 to 1995. A total of 1943 bronchoscopies were performed during this period, 47% in the HIV-positive group and 53% in the HIV negative group. Of 908 HIV-positive patients, 38 (4%) had a fungus isolated from the pulmonary sample whereas of 1035 HIV-negative patients, only 4 (0.2%) had a fungus isolated. Histoplasmosis and Cryptococcosis were more frequently found in HIV-positive than in HIV-negative patients (p < 0.001). Paracoccidioides brasiliensis was found in only 3 patients, all of them immunocompetent. The study demonstrated that, despite the low yields, the HIV-positive group may benefit from routine screening for fungal elements in specimens obtained by fiberoptic bronchoscopy. PMID- 10380565 TI - [Prevalence of intestinal helminthiasis in 100 municipalities in Venezuela (1989 1992)]. AB - A total of 113,254 individuals from 100 venezuelan municipalities were studied by mean of Kato-Katz coprological examination and the geohelminth prevalences were established. The national prevalences of T. trichiura, A. lumbricoides and Anquilostomideos were 32.6%, 26.9% and 5.6% respectively+. For T. trichiura the highest values were obtained for the following municipalities: Arevalo Gonzalez (54.6%) in Miranda state, Urama (76.9%) in Carabobo state and Punta de Piedra (78.4%) in Sucre state. For A. lumbricoides the highest values were determined in Punta de Piedra (63%), Tunapuy (61.9%) in Sucre state and Arevalo Gonzalez (62.7%) in Miranda state. For hookworm the highest values were detected in El Amparo (39.5%) and San Camilo (35.9%) in the state of Apure. In relation to the age, the highest prevalence of Ascariosis and Trichuriosis were determined in pre school and school children and for hookworm in adolescents and adults. A high variability between the municipalities belonging to the same state for the three geohelminths was observed. PMID- 10380566 TI - [Contribution to knowledge of reservoirs of Trypanosoma cruzi (Chagas, 1909) in Corrientes Province, Argentina]. AB - In order to identify Trypanosoma cruzi reservoirs in transmission areas, 60 mammals in Capital and San Luis del Palmar Departments, Corrientes, Argentina were studied. Primates, rodents, carnivores, marsupials and edentates were investigated, 40 of them living in captivity and 20 caught with traps in a rural area. The mammals were examined by xenodiagnosis and third or fourth instars nymphs of Triatoma infestans starved for 2 weeks were used. The feces were microscopically observed (400x) for Trypanosoma cruzi infection at 30, 60 and 90 days after feeding. Trypanosoma cruzi-like parasites were identified in 2 Saimiri sciureus and 1 Cebus apella analyzed by xenodiagnosis. It was concluded that parasitemia was low. However, the presence in a forest area of Didelphis albiventris, potential reservoir of the parasite, indicates a risk factor and deserves further epidemiological study for a true diagnosis of this endemic infection. PMID- 10380568 TI - Serum cytokines in chronic Chagas' disease. AB - We studied the serum levels of IL-2, IFN-gamma and TNF in different clinical forms of Chagas' disease and in patients clinically compensated and decompensated. Cytokines measured in 91 patients with the chronic form of the disease did not differ from those of 13 normal individuals, suggesting the absence of activation of the TH1 pattern of lymphocyte response. There were no statistical differences among the 17 patients in the indeterminate form of the disease, the patients presenting either early (n = 4) or well-developed signs of cardiomyopathy (n = 62), the digestive (n = 4) or the mixed (n = 4) forms of the disease. Serum TNF was undetectable and IFN-gamma levels did not differ between clinical forms and severities of Chagas' disease. However, we found IL-2 higher levels in the 25 non-controlled patients than in the 66 controlled individuals (p < 0.001). We suggest that IL-2 dosage may be useful as an indicator of the need for more aggressive procedures. PMID- 10380567 TI - [Trichome staining method applied to stools from HIV-infected patients with diarrhea, for microsporidia investigation]. AB - The microsporidia have been involved in several clinical manifestations in patients with AIDS, of whom diarrhoea is the commonest. The diagnosis of microsporidiasis depended on invasive procedures and the identification of the organisms is made by electron microscopy. The modified trichrome staining method allows that the diagnosis be made without such procedures by using light microscopy. In the present work, the modified trchrome method was applied in stools from 62 patients with diarrhoea, who had asymptomatic HIV infection or AIDS. Of the 62 samples analyzed, there was detection of microsporidial spores in one. This work confirms the existence of such protozoans in our patients, associated with manifestations of chronic diarrhoea in patients with AIDS who have severe immunodeficiency and ascertains that this staining method allows satisfactory identification of microsporidia from faeces, as well points out some directions to further studies. PMID- 10380569 TI - [Reactivation of Trypanosoma cruzi infection in patients with acquired immunodeficiency syndrome]. AB - A patient with AIDS and asymptomatic Chagas's disease and positive xenodiagnosis was taking ketoconazole in order to suppress parasitemia and prevent reactivation of Chagas's disease. Ketoconazole was unplanned suspended after 6 months, and the patient was admitted with fever, headache, vomiting, tachycardia, postural hypotension, hepatosplenomegaly, and positive xenodiagnosis one month later. Treatment with benzonidazole was begun leading to suppression of parasitemia. The patient had probability a neurotoxoplasmosis associated and progressed to coma and death with sepsis. No parasite was found in autopsy. PMID- 10380570 TI - Hepatitis B virus surface antigen (HBsAg) and antibody (anti-HBs) forming immune complexes in fulminant hepatitis. AB - This paper reports an unusual pattern of serological HBV markers and the presence of HBsAg/anti-HBs immune complexes in serum samples from two patients with fulminant hepatitis from the Brazilian Western Amazon Basin. The diagnosis was made by both serologic tests and demonstration of antigen/antibody complexes by transmission electron microscopy. Concurrent Delta virus superinfection is also discussed. PMID- 10380572 TI - Description of a possible clonal expansion of Plasmodium vivax in Manaus-Amazonas Brazil. AB - Atypical P. vivax cases reported in Manaus municipality led us to detect a genetic isolate of P. vivax. Variable regions of SSUrRNA were examined from the initial time of infection and in the two recrudescences/relapses from a patient exhibiting chloroquine and primaquine resistance. A unique isolate, found at all stages of infection, suggests the presence of a clonal expansion. PMID- 10380571 TI - The seroprevalence of hepatitis B and C in an Amerindian population in the southwestern Brazilian Amazon. AB - We have investigated the seroprevalence of hepatitis B and C among Karitiana Indians (n = 119) living in the State of Rondonia, southwestern Brazilian Amazon. The prevalences of anti-HBs and anti-HBc were 16.1% and 35.3%, respectively, with HBsAg being found in only four (3.4%) subjects. Anti-HCV antibodies were detected in two subjects (1.7%). Age-stratified prevalence data suggest that both vertical and horizontal (the last among adults) routes of HBV transmission are important in this community. PMID- 10380573 TI - [Culture development and morphological diagnosis of Emmonsia crescens in armadillos]. PMID- 10380574 TI - Controlling morbidity and interrupting transmission: twin pillars of lymphatic filariasis elimination. PMID- 10380576 TI - [Novel function of DNA polymerase in negative regulation of initiation of chromosomal replication]. PMID- 10380575 TI - [Ribosome recycling factor (RRF): a factor which disassemble the post-termination complex and reduces translational error]. PMID- 10380577 TI - [Commitment and differentiation of osteoclast on stromal cells]. PMID- 10380578 TI - [Genetic and molecular study on autophagy]. PMID- 10380579 TI - [Tight junctional permeability of intestinal epithelium and its regulation by food factors]. PMID- 10380580 TI - [Discovery of prolactin-releasing peptide in the brain]. PMID- 10380581 TI - [Producing clone animals: its historical background, present status and future scope]. PMID- 10380582 TI - [Nanometer/piconewton manipulation of single membrane protein molecules by laser tweezers]. PMID- 10380583 TI - [The fight for guidelines]. PMID- 10380584 TI - [Bioterrorism--threat and preparedness]. PMID- 10380585 TI - [Eclampsia]. PMID- 10380586 TI - [Different gender--equal treatment?]. PMID- 10380587 TI - [Chronic myeloid leukemia in health regions 1, 3, 4 and 5 during the period 1990 96]. AB - The aim of the present investigation was to obtain information about treatment, clinical course and outcome for all patients with chronic myeloid leukaemia through a six-year period in a defined part of Norway. A total number of 141 patients fulfilled the diagnostic criteria. This is equivalent to 0.9 patients per 100,000 per year. The median age was 62 years. More than 70% of the patients were primarily treated with hydroxyurea, either alone or combined with interferon. 40 out of 57 patients younger than 55 years underwent allogeneic stem cell transplantation. Median survival for all patients was 36 months with an estimated five-year survival rate of 33%. Patients older than 55 years had a median survival of 30 months with 16% alive after five years. The five-year survival rate for patients younger than 55 years was 56%, for transplanted patients 72%. 60 of 84 patients older than 55 years have died after 4 1/2 years median observation time. Two thirds of those died of leukaemia; one third of other causes. 23 of 57 patients younger than 55 years have died. 11 of them had had transplantations and most of them died from transplantation-related causes, while leukaemia was the dominating cause of death in the others. PMID- 10380588 TI - [Surgical treatment of endocrine ophthalmopathy]. AB - Thyroid ophthalmopathy is an inflammatory disorder of the extraocular muscles, orbital fat and orbital connective tissue that is most commonly seen in patients with Graves' hyperthyroidism. Inflammation is accompanied by deposition of extracellular matrix components, in particular glycosaminoglycans. The increase in the volume of the orbital contents may lead to periorbital swelling, extraocular muscle dysfunction, disfiguring proptosis, exposure keratitis, increased intraocular pressure and optic nerve compression. In many cases, surgical treatment is necessary for the rehabilitation of patients. In this report, we present a series of patients to illustrate relevant procedures and the results of surgical treatment in patients with thyroid ophthalmopathy. The records of all patients (66) with thyroid ophthalmopathy hospitalized in the Department of Ophthalmology, Haukeland University Hospital 1 April 1994-31 March 1998 were retrospectively evaluated. Orbital decompressions were performed in 43 patients (in 17 for compressive optic neuropathy), squint surgery in 13 patients, correction of eyelid retraction in 20 patients, and removal of excessive skin and fat from the eyelids in 11 patients. Average reduction of proptosis was 4 mm after lateral wall resection, and 6 mm after combined medial and lateral wall resection. Visual acuity improved in patients with compressive optic neuropathy to 6/6 or better in 18/20 eyes (postoperative data were not available for all patients), while that of the remaining two eyes was 6/9 and 6/24, respectively. Squint surgery was successful (no diplopia in primary or reading position) in eight patients after one procedure, and in four after two procedures. One patient has been scheduled for a third procedure due to a severe esotropia. In patients with thyroid ophthalmopathy, suboptimal treatment of the thyroid disorder may worsen the ophthalmopathy. 16 patients had their medication adjusted, ten were referred for thyroid surgery, and one for treatment with radioiodine. Treatment of patients with thyroid ophthalmopathy is a therapeutic challenge requiring close collaboration between different specialists. In severe cases, several surgical procedures may be needed. The complication rate is low, however, and for most patients the functional as well as the aesthetic situation is greatly improved. PMID- 10380589 TI - [Normal pressure hydrocephalus--evaluation of investigation procedures]. AB - 58 patients were investigated with a lumbar infusion test for normal pressure hydrocephalus in the department of neurology of the National Hospital between January 1991 and December 1996. We present a retrospective evaluation of the investigation protocol for these patients with suspected normal pressure hydrocephalus. The aim of the study was to find out whether specific prognostic factors could be identified by routine investigations. 20 patients had been referred to the department of neurosurgery and were subsequently shunted. A critical review of the criteria leading to surgical treatment (clinical symptoms, cerebral CT scan, infusion test, cisternography) is presented. There was no single variable which alone could discriminate normal pressure hydrocephalus from other types of hydrocephalus, or predict the outcome of shunting. A more systematic referral and clinical assessment of the patients together with a more precise evaluation of the findings and improved follow-up procedures are necessary for optimal selection of patients for surgery. The study also shows the importance of diagnosing these patients early, as delay in treatment appeared to worsen prognosis after shunting. PMID- 10380590 TI - [Choledochal cysts in adults]. AB - Based on a case report, we describe choledochal cysts in adults. Choledochal cysts are rare, and are often overlooked. The aetiology is unknown. The symptoms are pain or discomfort, episodes of jaundice, cholangitis and pancreatitis. Increased liver function tests and gallstone disease are common. There is an increased risk of malignant tumours in the cysts and adjacent organs. Ultrasonography, computer tomography, endoscopic retrograde cholangiopancreaticography, percutaneous cholangiography and magnetic resonance imaging with magnetic resonance cholangiopancreaticography are relevant diagnostic tools. Extrahepatic cysts should be operated with radical excision, with Roux-Y hepaticojejunostomy or other reconstructions allowing later diagnostic and therapeutic access to the bile ducts. Long term results are good. PMID- 10380591 TI - [Pneumatosis cystoides intestinalis]. AB - Patients with free intraperitoneal air usually undergo emergency surgery. Some of these patients will have no identifiable perforation, for instance those with pneumatosis cystoides intestinalis. This is a rare condition characterized by multiple intramural gas cysts in the gastrointestinal tract. The most common symptoms are meteorism, excessive flatulence, diarrhoea, abdominal pain, passage of mucus per rectum, or rectal bleeding. A case of pneumatosis cystoides intestinalsis is described. Plain abdominal radiographs showed distended bowel with free intraperitoneal air and intramural gas collections. At laparotomy, multiple intramural cysts were found, but no perforation or obstruction. The symptoms resolved after laparotomy, and the patient was discharged after a few days. The aetiology and pathogenesis of pneumatosis cystoides intestinalis are unknown, although deficient hydrogen metabolism and gasforming bacteria that penetrate the mucosal barrier may be involved. If needed, hyperbaric oxygen therapy is the treatment of choice. Surgery is indicated only in fulminant cases. PMID- 10380592 TI - [Serious bacterial and fungal infections in intravenous drug addicts]. AB - Invasive infections caused by bacteria and fungi are common complications of intravenous drug abuse. Various vital organs and structures may be affected, e.g. the cardiac valves, the larger arteries, the bones, the joints and the central nervous system. However, due to the high frequency of low-virulent microbes of skin and oral origin, the clinical picture may be atypical with subacute course and few focal signs and symptoms. The complexity of this problem is illustrated by eight cases of serious bacterial and fungal infections recently diagnosed at our hospitals. All patients were HIV negative intravenous heroin addicts. The clinical spectrum was wide and included skin abscesses, pyomyositis, spondylodiscitis, septic arthritis, costal osteomyelitis, infective endocarditis, recurrent bacteraemia, and multiple brain abscesses. PMID- 10380593 TI - [An outdoor person with high fever and flank pain]. PMID- 10380594 TI - [Surgical treatment of low back pain]. AB - Sciatica in low back pain is usually caused by lumbar disc herniation, degenerative spinal disease or lumbar spinal stenosis. Important pathogenetic factors are mechanical compression of and inflammatory changes in nerve roots caused by nucleus pulposus tissue. Decompression of nerve roots and removal of nucleus pulposus tissue are the main surgical goals. Emphasis should be placed on clinical identification of the symptomatic nerve roots. Computerized tomography (CT) and magnetic resonance (MR) imaging are the most important diagnostic tools today. Plain X-ray may be required for correct identification of the lowest mobile segment. Functional myelography combined with CT may be required to diagnose lumbar spinal stenosis. A proper selection of patients for surgery seems to be more important for a successful outcome than whether macro- or microsurgery is used, or wether the operation comprises one or several nerve roots. The combination of clear nerve root affection, mechanically provoced pain, and corresponding pathological findings at imaging are the best predictors of successful outcome. Psychosocial stability is an additional positive predictive factor. Impaired fibrinolysis occurring in smokers and in sedentary and obese patients, may be a negative predictive factor. PMID- 10380595 TI - [Radiological imaging in lumbago and sciatica]. AB - Low back pain and sciatica are among the most common medical problems in Western countries, affecting up to 80% of the population at some time during their lives. Plain radiography is still a sensitive method in degenerative spinal disease and for the identification of spondylolysis and destructions as well as transitional vertebra and other anomalies in the lumbosacral region. In lumbar disk herniation, CT and MR have higher sensitivity than lumbar myelography, and should be used as the primary imaging methods. Myelography is still the method of choice in lumbar spinal stenosis. Myelography should also be considered in patients with poor consistency between CT or MR findings and the clinical presentation. Postoperatively, MR is superior to CT and myelography for distinguishing between scar tissue and recurrent disk herniation. PMID- 10380597 TI - ["Uncertain" health complaints of women--a challenge for medicine and welfare state policy]. PMID- 10380596 TI - [Patients with chronic degenerative spinal disease--can conservative treatment reduce the waiting list for surgery?]. AB - Patients with a degenerative lumbar disorder selected for spinal fusion surgery by an experienced orthopaedic surgeon, were invited to participate in an exercise and behavioural modification programme. The goal was to let the patients experience that it is safe to move. 50 patients; mean age 49 years (19-76 years), mean duration of complaints ten years (2-39 years), agreed to participate. 26 patients had undergone back surgery. 80% were on sick leave or disability pension; 85% had either previously diagnosed psychiatric disorder or high scores for somatization or abnormal illness behaviour. At follow-up, 21 (42%) patients wanted surgery, 8 (16%) were uncertain, and 18 (36%) did not want surgery, 17 patients reported improved main symptoms, 18 were unchanged, and 12 had deteriorated. Previous surgery, illness behaviour and weak trunk muscles were associated with poor improvement and desire for surgery. Age, gender, emotional distress, intensity of pain, neurological or radiological signs or the number of exercise sessions taken did not influence results. PMID- 10380598 TI - [Clinical guidelines for primary health care]. PMID- 10380599 TI - [Zulu people and Isangoma people--HIV and tuberculosis]. PMID- 10380600 TI - [Influenza outbreaks in nursing homes]. PMID- 10380601 TI - [Negative report--how does it affect us?]. PMID- 10380602 TI - [For the 40th anniversary of the union of The 1st and The 2nd Moscow City Hospitals. Research schools of L.I. Sverzhevski'i, B.S. Preobrazhenski'i, V.T. Pal'chun]. PMID- 10380603 TI - [Paratonsillitis: aspects of therapeutic policy]. AB - A questionnaire has been distributed among ENT physicians in Russia and CIS on their approaches to management of paratonsillitis with reference to the disease stage. Many respondents approve active surgical policy and on demand perform abscesstonsillectomy in acute period. Those who perform the operations in "hot", "warm" and "cold" period fall into approximately equal groups. PMID- 10380604 TI - [Otogenic intracranial complications: current assessment of some aspects of the problem]. AB - Clinical cases and 245 case reports have been analyzed in terms of otogenic intracranial complications. Of late, there is a trend to lowering occurrence of otogenic complications and lethal outcomes compared to 1950-60. More favourable outcomes are attributed to introduction of new antibiotics and higher quality of ENT service. PMID- 10380605 TI - [Clinical picture and treatment of initial symptoms of otogenic intracranial complications]. AB - The author presents results of clinical and device investigation and analysis of intraoperative findings in surgical treatment of 150 patients with chronic purulent otitis media. Computed tomography of the temporal bones allows verification of the initial stage of otogenic intracranial complications in 90% of the cases. In early stages of development of otogenic intracranial complications destruction extention demans urgent cleansing intervention of the open type. PMID- 10380606 TI - [Complete and persistent cleansing: a major requirement for epitympanic surgeries]. AB - Because in chronic purulent epitympanitis pyodestruction affects bone tissues with resultant chronic osteomyelitis of the temporal bone it is thought necessary to remove radically all the foci of chronic inflammation and to open all the compartments of the middle ear in any operation for epitympanitis. This is also relevant to operations performed to create a small trepanation cavity or new sound conduction system. PMID- 10380608 TI - [The role of cervical proprioceptive afferentation in the mechanisms of vestibular dysfunction compensation. 2. The passive vestibulo-ocular reflex in bilateral labyrinth lesion]. AB - The study was made of passive cervicoocular reflex in patients with bilateral labyrinthine involvement. Passive cervicoocular reflex was assessed by three parameters: direction of the nystagmus, reactivity rate and phasic shift. It is proved that patients with bilateral labyrinthine lesions desplay intensive cervicoocular reflex serving the basic mechanism of vestibular dysfunction compensation in these patients. PMID- 10380609 TI - [Sub-atrophic and atrophic pharyngitis as a manifestation of dystrophic pharyngeal mucosa in presenile, senile and long living persons]. AB - The authors consider diagnostic, clinical and symptom characteristics of atrophic pharyngitis (AP) in presenile, senile and long-living persons and conclude that AP is age-specific dystrophic lesion of the pharynx. Diagnosis formulation by three degrees are suggested: pharyngeal dystrophy (PD) of degree I (only local symptoms cured after timely treatment), PD of degree II (local symptoms + neurotic condition which manifests with cancerophobia, persistent sensation of a foreign body), PD of degree III (persistent cancerophobia demanding psychotherapeutic treatment or psychiatric care). PMID- 10380610 TI - [On the differential diagnosis of median cysts of the neck]. AB - A case is reported of a 8-year-old girl with a median cyst of the neck treated surgically. Histologically, it was tuberculous lymphadenitis. It is recommended to account for peripheral lymph node tuberculosis in differential diagnosis of median cysts of the neck. PMID- 10380607 TI - [Correlation between clinical course and otitis media purulenta chronica and tonicity of autonomic nervous system]. AB - The study of the autonomic status of the body in 38 patients with different forms of chronic purulent otitis media (CPOM) has demonstrated that patients with exacerbation or complication of CPOM were for the most part sympathotonics and normotonics. The former had more persistent disease with more frequent recurrences. PMID- 10380611 TI - [Surgical treatment of congenital laryngeal cysts]. AB - Seven cases of congenital laryngeal cysts in newborns are analysed. The condition is proposed to be treated using an original endoscopic microsurgical technique designed by professor E.A. Tsvetkov. It enables early treatment with mobile laryngoscope. PMID- 10380612 TI - [Modern methods of nasal tip correction]. AB - The author reviews basic methods of nasal tip correction employed by current rhinosurgery. From 1994 to 1997 the methods were used in 112 plastic operations. Good and satisfactory results were achieved in 94.6% of the patients. PMID- 10380613 TI - [Combined polydex with phenylephrine preparations in the treatment of patients with nasal and paranasal inflammation]. PMID- 10380614 TI - [On rhinophyma and its cytological characteristics]. AB - Two cases of rhinophyma are described. Clinical and cytological features are specified. Rhinophyma can be diagnosed cytologically by hyperplastic cells of sebaceous glands. However, cytological diagnosis should also take into consideration clinical picture of the disease as the above cells can be encounted also in hyperplasia and/or adenoma of the sebaceous gland as well as basalioma with sebaceous differentiation. PMID- 10380615 TI - [Our experience with application of electrosurgical generator Force-30 in epistaxis management]. PMID- 10380616 TI - [A rare case of a large cholesteatoma of the paranasal sinuses]. PMID- 10380617 TI - [Congenital laryngeal cyst in a 3 month-old infant]. PMID- 10380619 TI - [Complications at anesthesia in tonsillectomy]. PMID- 10380618 TI - [A gunshot wound of the pharynx and spine]. PMID- 10380620 TI - Purification and characterization of D-glucosaminitol dehydrogenase from Agrobacterium radiobacter. AB - D-Glucosaminitol dehydrogenase, which catalyzes the conversion of D glucosaminitol to 3-keto-D-glucosaminitol, was purified to apparent homogeneity from extracts of Agrobacterium radiobacter. This organism has constitutively depressed levels of the enzyme but expression of the enzyme is induced by addition of D-glucosamine to the medium. Purification included ammonium sulfate fractionation and chromatography on columns of DEAE-Sephacel, Octyl-Sepharose CL 4B, and Cellulofine. The purified enzyme migrated as a single band, coinciding with dehydrogenase activities specific for D-glucosaminitol and ethanol, when electrophoresed on a 7.5% polyacrylamide gel at pH 8.0. Electrophoresis on a 12.5% PAGE in the presence of 1% SDS also yielded a single band. The enzyme had an apparent molecular mass of 79 kDa, as measured by the pattern of elution from a column of Cellulofine. The results indicated that the enzyme was a dimer of identical (or nearly identical) subunits of 39.5 kDa. D-Glucosaminitol dehydrogenase required NAD+ as a cofactor and used ethanol as the preferred substrate, as well as aliphatic alcohols with 2 to 4 carbon atoms, D glucosaminitol, D-glucosaminate, DL-allothreonine, glycerol, and erythritol as additional substrates. In 50 mM Tris-HCl buffer (pH 9.0) at 25 degrees C, the K(m) for D-glucosaminitol, ethanol, and NAD+ were 2.2, 2.0, and 0.08 mM, respectively. The enzyme had a pH optimum of 10 for D-glucosaminitol and 8.5 for ethanol. The enzyme lost substantial activity when treated with pyrazole, with certain reagents that react with sulfhydryl groups and with Zn2+ ion. The various results together suggest that the enzyme exploits different amino acid residues for the dehydrogenation of ethanol and of D-glucosaminitol. PMID- 10380621 TI - Characterization of yrpC gene product of Bacillus subtilis IFO 3336 as glutamate racemase isozyme. AB - Glr, the glutamate racemase of Bacillus subtilis (formerly Bacillus natto) IFO 3336 encoded by the glr gene, and YrpC, a protein encoded by the yrpC gene, which is located at a different locus from that of the glr gene in the B. subtilis genome, share a high sequence similarity. The yrpC gene complemented the D glutamate auxotrophy of Escherichia coli WM335 cells defective in the glutamate racemase gene. Glutamate racemase activity was found in the extracts of E. coli WM335 clone cells harboring a plasmid, pYRPC1, carrying its gene. Thus, the yrpC gene encodes an isozyme of glutamate racemase of B. subtilis IFO 3336. YrpC is mostly found in an inactive inclusion body in E. coli JM109/pYRPC1 cells. YrpC was solubilized readily, but glutamate racemase activity was only slightly restored. We purified YrpC from the extracts of E. coli JM109/pYRPC2 cells using a Glutathione S-transferase Gene Fusion System to characterize it. YrpC is a monomeric protein and contains no cofactors, like Glr. Enzymological properties of YrpC, such as the substrate specificity and optimum pH, are also similar to those of Glr. The thermostability of YrpC, however, is considerably lower than that of Glr. In addition, YrpC showed higher affinity and lower catalytic efficiency for L-glutamate than Glr. This is the first example showing the occurrence and properties of a glutamate racemase isozyme. PMID- 10380622 TI - Protective effects of dietary nasunin on paraquat-induced oxidative stress in rats. AB - The preventive effects of nasunin (delphinidin-3-[4-p-coumaroyl-rhamnosyl(1- >6)glucosid e]-5-glucoside) on paraquat-induced oxidative stress were determined in rats. Decreased food intake and body weight gain and increased lung weight by feeding the rats a diet containing paraquat were clearly suppressed by supplementing nasunin to the paraquat diet. Paraquat feeding increased the concentration of thiobarbituric acid-reactive substances (TBARS) in liver lipids and the atherogenic index, and decreased the liver triacylglycerol level. These effects were also suppressed by supplementing nasunin to the paraquat diet. In addition, catalase activity in the liver mitochondrial fraction was markedly decreased by feeding the paraquat diet, this decrease being partially suppressed by supplementing the paraquat diet with nasunin. These results suggest that nasunin acted preventively against the oxidative stress in vivo that may have been due to active oxygen species formed through the action of paraquat. PMID- 10380623 TI - Production and properties of the linamarase and amygdalase activities of Penicillium aurantiogriseum P35. AB - The effects of medium composition on the production of beta-glucosidase (amygdalase and linamarase) by Penicillium aurantiogriseum P35 were studied and the medium optimized as follows (g/l of deionized water): pectin, 10.0; (NH4)2SO4, 8.0; KH2PO4, 8.0; Na2HPO4, 2.8; MgSO4.7H2O, 0.5; yeast extract, 4.0; initial pH 6.0. When grown in a bench fermenter on this medium, the fungus produced 50.5 mU of amygdalase and 9.4 mU of linamarase per ml of culture broth. Two beta-glucosidases (PGI and PGII), each having amygdalase and linamarase activities, were recovered from the culture broth and purified; their relative molecular weights, as native enzymes, were estimated to be about 247,000 and 147,000, respectively. Both enzymes showed the same optimum pH (6.0) but different optimum temperatures (55 and 60 degrees C for PGI and PGII, respectively). Thermostability (10 min at 60 degrees C) and half-life of enzyme activity (7 hours at 60 degrees C) of PGII were higher than those of PGI (10 min at 50 degrees C and 2 hours at 55 degrees C, respectively). A wide range of cyanogenic glycosides (such as tetraphyllin B, epivolkenin, gynocardin, passibiflorin, prunasin, taxiphyllin, amygdalin, lucumin, sambunigrin, dhurrin, linamarin and cardiospermin sulfate) were hydrolyzed by both enzymes. PMID- 10380624 TI - Molecular cloning and expression of the gene encoding a phospholipase A1 from Aspergillus oryzae. AB - Phospholipase A1 (PLA1) is a hydrolytic enzyme that catalyzes removal of the acyl group from position 1 of lecithin to form lysolecithin. The genomic DNA and cDNA encoding PLA1 from Aspergillus oryzae were cloned with the mixed deoxyribonucleotide-primed polymerase chain reaction. The PLA1 gene is composed of 1,056 bp and has four exons and three short introns (63, 54, and 51 bp). The deduced amino acid sequence of PLA1 contained the N-terminal sequence of the mature PLA1 analyzed by Edman degradation. PLA1 cDNA has an open reading frame of 885 bp encoding the PLA1 precursor of 295 amino acid residues. The mature PLA1 is composed of 269 amino acid residues, and a prepro-sequence of 26 amino acid residues is at the N-terminal region of the PLA1 precursor. PLA1 has two possible N-glycosylation sites (Asn27 and Asn55). PLA1 has a consensus pentapeptide (-Gly His-Ser-Xaa-Gly-), which is conserved in lipases. The amino acid sequence of PLA1 showed 47% identity with that of mono- and diacylglycerol lipase from Penicillium camembertii. The PLA1 cDNA was expressed in Saccharomyces cerevisiae KS58-2D, indicating the cloned gene to be functional. PMID- 10380625 TI - The phylogeny of the cactophilic yeasts based on the 18S ribosomal RNA gene sequences: the proposals of Phaffomyces antillensis and Starmera caribaea, new combinations. AB - The complete sequences of the 18S rRNA gene fragments of the type strains of the cactophilic yeast species, Pichia antillensis, Pichia caribaea, Phaffomyces opuntiae, Phaffomyces thermotolerans, Starmera amethionina var. amethionina, and Starmera amethionina var. pachycereana were determined and compared. The type strain of Phaffomyces opuntiae had two kinds of the 18S rRNA gene sequences of which base differences were counted to be 15 and of which the percent similarity was calculated to be 99.1. The type strains of P. antillensis, P. caribaea, and Starmera amethionina var. pachycereana had the Q-7 system. The phylogenetic analyses showed that the genera Phaffomyces and Starmera were monophyletic and distant from each other and from the other species examined of the ascogenous teleomorphic genera, and that P. antillensis and P. caribaea were included within the clusters of the genera Phaffomyces and Starmera, respectively. The two Pichia species were transferred to the genera Phaffomyces and Starmera as the new combinations, Phaffomyces antillensis and Starmera caribaea. The new family Phaffomycetaceae was proposed as the type genus Phaffomyces. PMID- 10380626 TI - Modification of chitosan to improve its hypocholesterolemic capacity. AB - Cholestyramine is the most widely used bile acid sequestrant in the treatment of hypercholesterolemia. However, cholestyramine has unpleasant side effects as a consequence of its hydrophobic backbone. Therefore, high-capacity bile acid sequestering biopolymers with cationic chitosan derivatives were developed, because electrostatic interactions are important for binding with bile acid anions. Dialkylaminoalkylation and reductive amination of chitosan were done to add dialkylaminoalkyl and an additional free amino group at a hydroxyl site in the chitosan backbone respectively and the amino-derivatized chitosan derivatives were quaternized with methyl iodide to produce a cationic polyelectrolyte. The in vitro bile acid binding capacity of the chitosan derivatives in aqueous NaCl was measured by reversed-phase HPLC. The binding capacities of sodium glycocholate (a major bile acid) to chitosan, DEAE-chitosan, quaternized DEAE-chitosan, and cholestyramine were 1.42, 3.12, 4.06, and 2.78 mmol/g resin, respectively. With quaternized DEAE-chitosan, the bile acid binding capacity increased approximately 50% over that of cholestyramine. The bile acid binding capacity of dialkylaminoalkyl chitosan derivatives increased with the number of carbons in the alkyl groups, indicating that hydrophobic interaction is a secondary factor for the sequestration of bile acids. PMID- 10380627 TI - Construction of an effective host-vector system for the yeast Saccharomyces exiguus Yp74L-3. AB - An effective host-vector system specific to the yeast Saccharomyces exiguus Yp74L 3 was constructed to promote the molecular genetic analyses for the yeast. To obtain a stable reversionless host strain, we constructed an S. exiguus strain carrying leu2::ScURA3 by disrupting the S. exiguus LEU2 gene with the S. cerevisiae URA3 gene. A vector plasmid unique to S. exiguus was subsequently developed by inserting both the LEU2 gene and an ARS cloned from S. exiguus into an Escherichia coli phagemid, pUC119. The vector constructed, pTH119 was able to transform the S. exiguus leu2::ScURA3 strain to Leu+ efficiently. The stability of the vector in the S. exiguus host cells resembled that of a YRp-type vector in S. cerevisiae. PMID- 10380628 TI - Cloning of a gene encoding hydroxyquinol 1,2-dioxygenase that catalyzes both intradiol and extradiol ring cleavage of catechol. AB - Two Escherichia coli transformants with catechol 1,2-dioxygenase activity were selected from a gene library of the benzamide-assimilating bacterium Arthrobacter species strain BA-5-17, which produces four catechol 1,2-dioxygenase isozymes. A DNA fragment isolated from one transformant contained a complete open reading frame (ORF). The deduced amino acid sequence of the ORF shared high identity with hydroxyquinol 1,2-dioxygenase. An enzyme expressed by the ORF was purified to homogeneity and characterized. When hydroxyquinol was used as a substrate, the purified enzyme showed 6.8-fold activity of that for catechol. On the basis of the sequence identity and substrate specificity of the enzyme, we concluded that the ORF encoded hydroxyquinol 1,2-dioxygenase. When catechol was used as a substrate, cis,cis-muconic acid and 2-hydroxymuconic 6-semialdehyde, which were products by the intradiol and extradiol ring cleavage activities, respectively, were produced. These results showed that the hydroxyquinol 1,2-dioxygenase reported here was a novel dioxygenase that catalyzed both the intradiol and extradiol cleavage of catechol. PMID- 10380629 TI - Potassium/proton antiport system is dispensable for growth of Enterococcus hirae at low pH. AB - An energy-dependent K+/H+ antiport system is found in Enterococcus hirae ATCC 9790 cultured in a standard complex medium (Y. Kakinuma, and K. Igarashi, J. Biol. Chem. 263:14166-14170, 1988). We have now found that the activity of this antiport system was totally missing in cells cultured in a defined medium. In this defined medium, E. hirae did not grow well at pH near 9, but grew normally at pH below 7.5. This antiport system is important at high pH but dispensable at lower pH for ion homeostasis of this bacterium. PMID- 10380630 TI - Assembly of Staphylococcus aureus leukocidin into a pore-forming ring-shaped oligomer on human polymorphonuclear leukocytes and rabbit erythrocytes. AB - Staphylococcal leukocidin consists of two separate proteins, LukS and LukF, which cooperatively lyse human and rabbit polymorphonuclear leukocytes and rabbit erythrocytes. Here we studied the pore-forming properties of leukocidin and the molecular architecture of the leukocidin pore. (1) Leukocidin caused an efflux of potassium ions from rabbit erythrocytes and swelling of the cells before hemolysis. However, ultimate lysis of the toxin-treated swollen erythrocytes did not occur when polyethylene glycols with hydrodynamic diameters of > or = 2.1 nm were present in the extracellular space. (2) Electron microscopy showed the presence of a ring-shaped structure with outer and inner diameters of 9 and 3 nm, respectively, on leukocidin-treated human polymorphonuclear leukocytes and rabbit erythrocytes. (3) Ring-shaped structures of the same dimensions were isolated from the target cells, and they contained LukS and LukF in a molar ratio of 1:1. (4) A single ring-shaped toxin complex had a molecular size of 205 kDa. These results indicated that LukS and LukF assemble into a ring-shaped oligomer of approximately 200 kDa on the target cells, forming a membrane pore with a functional diameter of approximately 2 nm. PMID- 10380631 TI - A competitive enzyme-linked immunosorbent assay-like method for the measurement of urinary hyaluronan. AB - A highly sensitive method for measurement of urinary hyaluronan with a minimum molecular mass of 2,000 Da was developed without using HPLC or radioisotopes. This competitive enzyme-linked immunosorbent assay-like method, used competitive binding of free hyaluronan in the sample and biotin-labeled standard hyaluronan to hyaluronan binding protein in solid phase. A total of 150 healthy individuals from both sexes at ages from 0 to 100 years was examined by the established method. Hyaluronan of 384 +/- 80 ng/mg creatinine (mean +/- SD) was constantly excreted into urine of 24-40-year-old healthy adults. The urinary hyaluronan levels were significantly higher before age 1 (p < 0.001) and rather high after 90 years compared to the other groups. The average molecular weight of urinary hyaluronan (5,500 Da) was constant through all generations. Sex difference of urinary hyaluronan was not observed both quantitatively or qualitatively. PMID- 10380632 TI - Antiproliferative activity of flavonoids on several cancer cell lines. AB - Twenty-seven Citrus flavonoids were examined for their antiproliferative activities against several tumor and normal human cell lines. As a result, 7 flavonoids were judged to be active against the tumor cell lines, while they had weak antiproliferative activity against the normal human cell lines. The rank order of potency was luteolin, natsudaidain, quercetin, tangeretin, eriodictyol, nobiletin, and 3,3',4',5,6,7,8-heptamethoxyflavone. The structure-activity relationship established from comparison among these flavones and flavanones showed that the ortho-catechol moiety in ring B and a C2-C3 double bond were important for the antiproliferative activity. As to polymethoxylated flavones, C 3 hydroxyl and C-8 methoxyl groups were essential for high activity. PMID- 10380633 TI - Enhancing effect of dietary vinegar on the intestinal absorption of calcium in ovariectomized rats. AB - We studied the effect of dietary vinegar on calcium absorption by using ovariectomized rats fed on a low-calcium diet. The apparent absorption of calcium was higher when the rats were fed on a diet containing 1.6% vinegar for 32 days than when fed on a diet without vinegar (P < 0.05). The calcium content in the femur of the rats given diets containing 0.4% and 1.6% vinegar were also higher (P < 0.05). The serum parathyroid hormone level was lower and the crypt depth of the duodenum thicker in the rats fed on a diet containing 1.6% vinegar (P < 0.05). These results suggest that dietary vinegar enhanced intestinal calcium absorption by improving calcium solubility and by the trophic effect of the acetic acid contained in vinegar, which would reduce the bone turnover caused by ovariectomy and be helpful in preventing osteoporosis. PMID- 10380634 TI - Localization of sequential antigenic determinants on bovine beta-casein with synthetic peptides and antisera from mouse, rabbit, and goat. AB - The antigenic determinants of bovine beta-casein (beta-CN) were localized by using twenty overlapping peptides encompassing the entire sequence of beta-CN and anti-beta-CN antisera from outbred mouse, rabbit and goat. The profile of the reactions was characteristic to the species, the dominant antigenic regions being 80-95, 143-158 and 195-209 in mouse, 1-16 in rabbit and 100-115 in goat. Regions 1-16, 100-115, 121-136 and 143-158 were antigenic in all three species. The number of antigenic regions recognized by goat was much fewer than that by mouse and rabbit, possibly because of the homology between bovine and goat beta-CN. A mixture of the twenty peptides could absorb about 50-60% of beta-CN specific antibodies from each species. Furthermore, the mouse and rabbit anti-beta-CN antibodies were also specific to the phosphorylated regions. We therefore conclude that the major antigenic determinants on beta-CN would be largely sequential and include the phosphorylated sites. PMID- 10380635 TI - Two isoforms of a human actin-related protein show nuclear localization and mutually selective expression between brain and other tissues. AB - Actin-related proteins (Arps), which are divergent, but apparently homologues to actin, are categorized into 10 classes. While Arps belonging to classes 1-3 were found to be localized in the cytoplasm across eukaryotic phyla, other classes of Arps were found mostly in invertebrates and suggested to contribute to structural modulation of chromatin. Here we report the identification and the characterization of two human isoforms of an Arp not belonging to classes 1-3, which we designated hArpN alpha and hArpN beta. Both proteins were expressed in HeLa cells and they were found localized within the nucleus. Most interestingly, in different human tissues, hArpN alpha and beta were found to be expressed mutually exclusively, and the expression of hArpN alpha was absolutely restricted to the brain. These findings suggest that, in vertebrates, members of distantly related Arps might have tissue-specific functions in the nucleus, possibly through structural modulation of chromatin. PMID- 10380636 TI - Multi-enzymatic glucosylation using Eucalyptus UDP-glucosyltransferase coupled UDPglucose-fermentation by bakers' yeast. AB - The enzymatic synthesis of glucoside compounds using a membrane-associated UDP glucosyltransferase fraction from Eucalyptus perriniana cultured cells as a water insoluble catalyst (N. Nakajima, et. al., J. Ferment. Bioeng., 84 (5), pp. 455 460, 1997) has been effectively done by coupling UDPglucose-fermentation by bakers' yeast. For example, beta-thujaplicin (hinokitiol) and p-aminobenzoic acid were converted respectively to their corresponding beta-D-monoglucosides with the conversion rate of around 24-26% by the multi-enzymatic system with UDPglucose as a glucose donor, which is produced by yeast cells from glucose and 5'-UMP. Addition of either cellobiose, a substrate of beta-glucosidase, or DL-1,2-anhydro myo-inositol, an inhibitor for the enzyme in the reaction mixture, could increased the yield of these beta-D-monoglucosides. This new enzymatic system could also be used for the synthesis of flavonoid glucosides such as isoquercitrin (quercetin 3-O-beta-D-glucoside). PMID- 10380637 TI - cDNA sequence analysis of a novel member of the three loop protein family from the Chinese continental banded krait. AB - The cDNA encoding a novel three loop protein was cloned from cellular RNA isolated from the venom gland of Bungarus multicinctus multicinctus by RT-PCR. The mature protein has 82 amino acid residues. It shared only 25-38% similarity with some cardiotoxins and did not have sequence similarity with neurotoxins, while its cDNA was about 70% similar to both the cDNAs encoding neurotoxins and the cDNAs encoding cardiotoxins. PMID- 10380638 TI - Production of rhamnolipid biosurfactant by fed-batch culture of Pseudomonas aeruginosa using glucose as a sole carbon source. AB - The pH-stat fed-batch culture of Pseudomonas aeruginosa YPJ-80 was done to produce a rhamnolipid biosurfactant. With glucose as the sole carbon source, the final concentrations of cells and rhamnolipid biosurfactant obtained in 25 h were 25 g cell dry weight/l and 4.4 g/l, respectively. PMID- 10380640 TI - Inhibition of iron/ascorbate-induced lipid peroxidation by an N-terminal peptide of bovine lactoferrin and its acylated derivatives. AB - Bovine lactoferrin (LF) and lactoferricin B (LFcin B), an antimicrobial peptide derived from bovine LF, inhibited thiobarbituric acid-reactive substance (TBARS) formation in a iron/ascorbate-induced liposomal phospholipid peroxidation system. The inhibition of TBARS formation occurred with N-acylated 9-mer peptides with a core sequence of LFcin B and, compared to LFcin B, their antioxidant effect was clearly observed at a concentration almost 100 times lower. PMID- 10380642 TI - Isolation and structural assignment of 5-deoxytetrodotoxin from the puffer fish Fugu poecilonotus. AB - A novel 10,7-lactone type of tetrodotoxin analog, 5-deoxytetrodotoxin, was isolated from the puffer fish, Fugu poecilonotus, and its structure was assigned by spectroscopic methods. PMID- 10380643 TI - Laboratory evaluation of crude oil biodegradation with commercial or natural microbial inocula. AB - Experiments have been performed to screen eight microbial commercial products that, according to the manufacturers, are able to degrade crude oil. This study compared the crude oil biodegradation activity of commercial inocula with that of natural inocula (activated sludge and tropical aquarium water). Some of the latter were previously adapted to the crude oil as the only carbon source. Nutrients and sorbents in the commercial formulations were eliminated, and each inoculum was precultured on marine yeast extract medium. Crude oil biodegradability tests were conducted with close initial substrate concentration to initial bacterial concentration ratios (S0/X0) of 0.94 g of crude oil/10(9) CFU, which allowed a comparison of biodegradation activity. The inocula oxidized the crude oil after a short lag time of less than 3-18 days. After that time, the rate of oxidation varied between 45 and 244 mg O2/(L.day). Crude oil biodegradation after a 28-day test was effective only for 10 out of 12 inocula (from 0.1 to 25% in weight). Biodegradation mainly corresponded to the saturated fraction of the crude oil; the asphaltene fraction was never significantly biodegraded. Our results led to the conclusion that natural inocula, either adapted or not adapted to crude oil, were the most active (from 16 to 25% of loss in crude oil weight) and only one commercial inoculum was able to degrade 18% of the crude oil. Other inocula had a biodegradation activity ranging from 0.1 to 14%. PMID- 10380644 TI - Characterization of metal-resistant soil eubacteria by polymerase chain reaction- denaturing gradient gel electrophoresis with isolation of resistant strains. AB - Contamination of soils with heavy metal ions is a major problem on industrial and defense-related sites worldwide. The bioavailability and mobility of these contaminants is partially determined by the microbial biomass present at these sites. In this study, we have assessed the effect of the addition of a mixture of toxic metal salts on the prokaryotic community of microcosms consisting of sandy loam soil using direct molecular analysis of the recoverable eubacterial 16S rDNA molecules by polymerase chain reaction--denaturing gradient gel electrophoresis (PCR-DGGE) and limited phospholipid fatty acid analysis (PLFA). Addition of toxic metals (nonradioactive surrogates of Sr, Co, Cs, Cd) resulted in rapid (ca. 1 week) changes in the DGGE profile of the indigenous eubacterial community when compared with pristine controls. These changes were stable over the course of the experiment (8 weeks). No changes in the eubacterial population of control microcosms were detected. The major changes in community structure in metal contaminated microcosms consisted of the appearance of four novel bands not detected in controls. Sequence analysis of these bands suggested that two organisms related to the genus Acinetobacter and two related to the genus Burkholderia carried a selective advantage over other indigenous eubacteria under heavy metal induced stress. The Burkholderia spp. were then cultured and further characterized using lipid analysis. PMID- 10380645 TI - The role of catalase in hydrogen peroxide resistance in fission yeast Schizosaccharomyces pombe. AB - The role of catalase in hydrogen peroxide resistance in Schizosaccharomyces pombe was investigated. A catalase gene disruptant completely lacking catalase activity is more sensitive to hydrogen peroxide than the parent strain. The mutant does not acquire hydrogen peroxide resistance by osmotic stress, a treatment that induces catalase activity in the wild-type cells. The growth rate of the disruptant is not different from that of the parent strain. Additionally, transformed cells that overexpress the catalase activity are more resistant to hydrogen peroxide than wildtype cells with normal catalase activity. These results indicate that the catalase of S. pombe plays an important role in resistance to high concentrations of hydrogen peroxide but offers little in the way of protection from the hydrogen peroxide generated in small amounts under normal growth conditions. PMID- 10380646 TI - Influence of chemical surfactants on the biodegradation of crude oil by a mixed bacterial culture. AB - The effects of surfactant physicochemical properties, such as the hydrophile lipophile balance (HLB) and molecular structure, on the biodegradation of 2% w/v Bow River crude oil by a mixed-bacterial culture were examined. Viable counts increased 4.6-fold and total petroleum hydrocarbon (TPH) biodegradation increased 57% in the presence of Igepal CO-630, a nonylphenol ethoxylate (HLB 13, 0.625 g/L). Only the nonylphenol ethoxylate with an HLB value of 13 substantially enhanced biodegradation. The surfactants from other chemical classes with HLB values of 13 (0.625 g/L) had no effect or were inhibitory. TPH biodegradation enhancement by Igepal CO-630 occurred at concentrations above the critical micelle concentration. When the effect of surfactant on individual oil fractions was examined, the biodegradation enhancement for the saturate and aromatic fractions was the same. In all cases, biodegradation resulted in increased resin and asphaltene concentrations. Optimal surfactant concentrations for TPH biodegradation reduced resin and asphaltene formation. Chemical surfactants have the potential to improve crude oil biodegradation in complex microbial systems, and surfactant selection should consider factors such as molecular structure, HLB, and surfactant concentration. PMID- 10380647 TI - Isolation of Desulfovibrio intestinalis sp. nov. from the hindgut' of the lower termite Mastotermes darwiniensis. AB - A Gram-negative, anaerobic sulfate-reducing bacterium was isolated from hindgut contents of the lower termite Mastotermes darwiniensis Froggatt (strain KMS2). Strain KMS2 is motile by a single polar flagellum. The isolate possesses desulfoviridin and catalase activity. The G+C content of its DNA is in the range of 54.5-55.5 mol% (strain KMS2). It respires hydrogen and different low molecular weight organic compounds in the presence of sulfate, thiosulfate, and sulfite, and also oxygen. The isolated strain ferments pyruvate. Fastest growth with a doubling time of 12.5 h was obtained at 37 degrees C and not at 28 degrees C, the temperature at which the termites were grown. The isolate showed a 16S rDNA sequence homology of 95.9% to Desulfovibrio desulfuricans ATCC 27774 and a DNA DNA homology of 44.6% to D. desulfuricans Essex 6 (type strain). Based on its biochemical properties and 16S rDNA sequence, the isolate was assigned to a new species named Desulfovibrio intestinalis. PMID- 10380648 TI - Mutant analysis of Prevotella sp. plaA-lacZ fusion protein expression in Escherichia coli: support for an essential role of the stem-loop. AB - This study investigated the involvement of RNA folding in the synthesis of a fusion protein with beta-galactosidase activity. The coding gap region of the Prevotella loescheii adhesin gene plaA was fused in-frame with the Escherichia coli lacZ gene on plasmid pSK105. N-Terminal sequencing of the expressed plaA lacZ protein indicated that it resulted from translational initiation at a fortuitous ribosomal-binding site within the plaA sequence at nt 570. Specific mutations were introduced in the stem-loop region that precedes the gap sequence. Analysis of stem-loop mutants, together with the introduction of compensatory mutations that restored activity, supports a requirement for stem-loop formation within the plaA sequence preceding the translational initiation site. A mutation reducing the predicted size of the loop, but preserving the stem structure, inactivated fusion protein synthesis. A suppressor mutation predicted to restore the size of the loop restored efficient fusion protein synthesis. In addition, the sequence preceding the translational start site of the plaA-lacZ fusion has several similarities to sequences that function as translational enhancers in prokaryotes. These include a stem-loop structure, an A-U rich region preceding the initiation codon, and a region of homology to 16S rRNA. PMID- 10380649 TI - Phenol degradation by an enterobacterium: a Klebsiella strain carries a TOL-like plasmid and a gene encoding a novel phenol hydroxylase. AB - Although phenol catabolism is described for many different microorganisms, there is no example for such a pathway in an enterobacterial strain. Here we characterize a Klebsiella oxytoca strain that grows on phenol as the only source of carbon and energy. As the key enzyme of phenol degradation, phenol hydroxylase was purified to apparent homogeneity. Compared with other phenol hydroxylases, the Klebsiella enzyme differs with respect to several properties: (i) SDS-PAGE and gel-filtration analysis of the purified protein revealed that the enzyme is a monomer with a molecular mass of 156 kDa; (ii) steady-state kinetic measurements resulted in a K(m) value of 0.22 mM for phenol; and (iii) the enzyme is both dependent on NADPH/FAD and sensitive to EDTA. Further degradation of catechol, the reaction product of phenol hydroxylase, may occur via the effective meta fission pathway often located on TOL or TOL-like plasmids. Such a plasmid was prepared from the Klebsiella strain and further characterized. The given data demonstrate that the isolated strain exhibits all characteristics of an efficient phenol-degrading microorganism. PMID- 10380650 TI - Kodamaea kakaduensis and Candida tolerans, two new ascomycetous yeast species from Australian Hibiscus flowers. AB - Two new yeast species were isolated from flowers of Hibiscus species in Eastern and Northern Australia. Kodamaea kakaduensis is heterothallic, haploid, and similar to other Kodamaea species and to Candida restingae. Buds are often produced on short protuberances, and a true mycelium is formed. The new species differs from others by the assimilation of trehalose, melezitose, and xylitol, and is reproductively isolated. The cells of Candida tolerans are small and a pseudomycelium is formed. The carbon and nitrogen assimilation pattern is reminiscent of that of Zygosaccharomyces rouxii but the two are not closely related. Sequences of the D1/D2 domain of large subunit ribosomal DNA confirm the membership of K. kakaduensis in the genus Kodamaea and indicate that C. tolerans belongs to the Clavispora-Metschnikowia clade, with a moderate relatedness to Candida mogii. The type strains are: K. kakaduensis, UWO(PS)98-119.2 (h+, holotype, CBS 8611) and UWO(PS)98-117.1 (h-, isotype, CBS 8612); and C. tolerans, UWO(PS)98-115.5 (CBS 8613). PMID- 10380651 TI - Pyruvate metabolism by Anaplasma marginale in cell-free culture. AB - Partially purified Anaplasma marginale initial bodies were cultivated in a cell free system in the presence of [3-14C]pyruvate for 24 or 48 h. Experiments showed that a significant portion of the pyruvate supplied to the cultures was incorporated into initial body components. Label incorporation was reduced by 72% in the presence of oxytetracycline. Fractionation and chromatography of the organisms revealed radioactive incorporation as alanine. This is the first report of de novo amino acid synthesis by A. marginale demonstrating that the rickettsia is capable of using pyruvate, an erythrocyte glycolytic product, in its metabolism. PMID- 10380652 TI - Purification and characterization of the western spruce budworm larval midgut proteinases and comparison of gut activities of laboratory-reared and field collected insects. AB - Three proteolytic enzymes, trypsin, chymotrypsin, and aminopeptidase-N (APN), were purified from laboratory-reared western spruce budworm, Choristoneura occidentalis [Freeman], larvae. Budworm trypsin exhibited a high degree of substrate specificity, was inactivated by DFP and TLCK, and was inhibited by trypsin inhibitors. The western spruce budworm chymotrypsin hydrolyzed SAAPFpNA and SAAPLpNA, but not SFpNA, SGGFpNA, SGGLpNA or BTpNA. The chymotrypsin was inactivated by DFP, and was inhibited by chymostatin and the chymotrypsin inhibitor, POT-1. Purified budworm chymotrypsin exhibited little BTEE esterolytic activity and was insensitive to inhibition with TPCK. The N-terminal sequence of budworm trypsin, chymotrypsin, and APN were obtained. Similar levels of trypsin and APN gut activities were found in laboratory-reared and field-collected larvae. However, in comparison to laboratory-reared insects, considerably less chymotrypsin activity, and a much higher level of gut carboxypeptidase activity were found in field-collected western spruce budworm larvae. PMID- 10380653 TI - Differential mRNA expression levels and gene sequences of a putative carboxylesterase-like enzyme from two strains of the parasitoid Anisopteromalus calandrae (Hymenoptera: Pteromalidae). AB - Carboxylesterase-like enzyme cDNAs have been cloned and sequenced from malathion resistant and susceptible strains of the parasitoid Anisopteromalus calandrae (Howard) (Hymenoptera: Pteromalidae). The cDNAs consist of 1963 nucleotides including a 35 bp untranslated 5'-end, a 1596 bp open reading frame, and a 332 bp untranslated 3'-end. The open reading frame encodes 532 amino acid residues. The predicted protein sequence from these cDNAs includes 2 potential N-glycosylation sites, a carboxylesterase type-B serine active site FGGDSENVTIFGESAG, and conserved residues Ser187, Glu317, and His432 to function as the catalytic triad. The predicted carboxylesterase-like enzyme sequence is most similar to that of the carboxylesterase from the peach-potato aphid, Myzus persicae with 45% sequence identity. Alignment of the parasitoid carboxylesterase-like enzyme cDNAs revealed that there are two nucleotide differences in the open reading frame between the parasitoid strains, including a silent mutation and a point mutation that presumably causes a gene product difference. A nucleotide thymine at position 658 in the susceptible strain cDNA is replaced by a guanine in the resistant strain cDNA. This substitution leads to an amino acid change from tryptophan (Trp220) in the susceptible strain to glycine (Gly220) in the resistant strain. This substitution is genetically linked to resistance but it is not known how or if this amino acid substitution affects detoxification of malathion. Northern blot analyses demonstrated that expression level of the carboxylesterase-like enzyme mRNA in adult A. calandrae is approximately 30-fold higher in the resistant strain relative to that in the susceptible strain. Southern analysis indicated that Pst I or Eco RI restriction sites are different in the two strains. Both a modified gene structure and an increase in expression of carboxylesterase may be responsible for the high level of resistance found in this beneficial wasp. PMID- 10380654 TI - Molecular characterization of MRJP3, highly polymorphic protein of honeybee (Apis mellifera) royal jelly. AB - Major proteins of honey bee (Apis mellifera) royal jelly are members of the MRJP protein family. One MRJP protein termed MRJP3 exhibits a size polymorphism as detected by SDS-PAGE. In this report we show that polymorphism of the MRJP3 protein is a consequence of the polymorphism of a region with a variable number of tandem repeats (VNTR) located at the C-terminal part of the MRJP3 coding region. We present the characterization of five polymorphic alleles of MRJP3 by DNA sequencing. By PCR analyses, at least 10 alleles of distinct sizes were found in randomly sampled bees. Studies with nurse bees from a single honeybee colony revealed both Mendelian inheritance and very high variability of the MRJP3 genomic locus. The high variability and simple detection of the MRJP3 polymorphism may be useful for genotyping of individuals in studies of the honeybee. PMID- 10380655 TI - Cloning and functional expression of a cDNA encoding a metabolic acyl-CoA delta 9 desaturase of the cabbage looper moth, Trichoplusia ni. AB - Acyl-CoA delta 9-desaturases play essential roles in fatty acid metabolism and the regulation of cell membrane fluidity. In this research, a cDNA sequence was obtained from Trichoplusia ni adult fat body mRNA by using RT-PCR with degenerate primers based on other characterized delta 9-desaturase sequences. The remainder of the sequence was amplified using 3'- and 5'-RACE. A 1439 bp cDNA reconstructed from three overlapping PCR products contains an ORF encoding a 353-amino acids (aa) protein that shows clear homology (greater than 50% aa identity and greater than 65% aa similarity to characterized insect and vertebrate desaturases). The ORF of this cDNA was subcloned into an expression vector, which relieved the unsaturated fatty acid (UFA) auxotrophy of a desaturase-deficient yeast strain following genetic transformation. The newly characterized desaturase from T. ni produced fatty acids delta 9-16 and delta 9-18 in a 1:6 ratio, compared to a 5:1 ratio, respectively, with the yeast delta 9 desaturase. A Northern blot hybridization and a RT-PCR experiment showed that temporal and tissue-specific patterns of expression of the corresponding mRNA are distinct from those of the delta 11-desaturase mRNA present in the pheromone glands of adult females. Based on its homology to other desaturases, the widespread distribution of its corresponding mRNA in various tissues, and its functional assay, we conclude that this cDNA encodes the apoprotein corresponding to the desaturase component of the metabolic delta 9-desaturase complex of T. ni. PMID- 10380656 TI - Prolonged negative selection of Drosophila melanogaster for a character of adaptive significance disturbs stress reactivity. AB - The metabolism of juvenile hormone by JH-esterase and JH-epoxide hydrolase, and octopamine by tyrosine decarboxylase were studied under normal and stress conditions in flies of two related lines of D. melanogaster. One was selected for high (HA line) and another for low (LA line) male sexual activity for more than 700 generations. It was demonstrated that prolonged selection for low male sexual activity results in considerable changes in both systems. Tyrosine decarboxylase activity in males and females of the LA line was sharply reduced as compared with those of the HA and control Canton-S lines; JH-esterase and JH-epoxide hydrolase activities were decreased in females, and not in males, of the LA line. It was demonstrated that the response of both metabolic systems to heat stress is impaired in individuals of the LA line: the system of juvenile hormone metabolism does not respond to stress, and that of octopamine metabolism is decelerated. The role of juvenile hormone metabolism in male reproductive function is discussed. PMID- 10380657 TI - A maternal gene mutation correlates with an ovary phenotype in a parthenogenetic wasp population. AB - Endoparasitoid wasps rely on maternal protein secretions, including viruses and virus-like particles (VLPs), to overcome host defense reactions. In the ichneumonid Venturia canescens, VLPs are assembled in the nuclei of ovarian calyx gland cells, secreted into the lumen of the gland, and eventually transmitted into the host caterpillar together with the parasitoid egg. One of the genes coding for VLP proteins, termed VLP1, exists in two alleles producing two structurally different proteins. Here we describe the establishment and initial phenotypic characterisation of two parthenogenetic laboratory strains, which differ in VLP1 as well as in other genetic markers. A comparison of calyx tissues from the two strains revealed morphological differences that seem to affect egg movement from the ovarioles into the oviduct. The observed histological changes are correlated with differences in egg maturation and embryonic development causing a delay in larval hatching in one of the strains. Under conditions that favour superparasitism, the two strains differ in the number of offspring produced. PMID- 10380658 TI - Isolation of a cDNA for an octopamine-like, G-protein coupled receptor from the cattle tick, Boophilus microplus. AB - Octopamine is a biogenic amine neurotransmitter of invertebrates that binds to a G-protein coupled receptor that has seven transmembrane domains. Formamidine pesticides like amitraz are highly specific agonists of the octopamine receptor. Amitraz is used extensively to control the cattle tick, Boophilus microplus, and many other ticks but now there are strains of ticks that are resistant to amitraz. We have isolated a cDNA from the cattle tick, B. microplus, that belongs to the biogenic amine family of receptors. The predicted amino acid sequence from this cDNA is most similar to octopamine receptors from insects. The nucleotide sequence of this gene from amitraz-resistant and amitraz-susceptible cattle ticks was identical. Thus, a point mutation/s did not confer resistance to amitraz in the strains we studied. Alternative explanations for resistance to amitraz in B. microplus are discussed. PMID- 10380659 TI - Metalinguistic awareness in children: a developmental progression. AB - The purpose of this study was to determine if a developmental order exists in the metalinguistic ability of children to make judgments about the form of language while simultaneously attending to a meaningful linguistic context. The stimulus material consisted of a short story into which 20 nonsense lexical items had been substituted. The 20 stimuli were comprised of phonotactically illegal and legal sequences of phonemes. In addition, the lexical items had been positioned to replace either structure or content words within the story. The participants were 90 Caucasian children who were divided into nine age groupings from 4;0 to 12;11. Baseline data were obtained from 10 adults. All subjects were required to respond to the audio-recorded stimuli by pressing a button whenever a nonsense item was perceived. The data were analyzed for both number of correct responses and reaction times. Results revealed a major shift in metalanguage ability occurring between 7 and 8 years of age. The 8- to 12-year-olds responded correctly to more items and at significantly faster rates than the 4- to 7-year-olds. The adults outperformed the children on all tasks, showing that metalanguage development continues beyond childhood. PMID- 10380660 TI - Emotion in speech: the acoustic attributes of fear, anger, sadness, and joy. AB - Decoders can detect emotion in voice with much greater accuracy than can be achieved by objective acoustic analysis. Studies that have established this advantage, however, used methods that may have favored decoders and disadvantaged acoustic analysis. In this study, we applied several methodologic modifications for the analysis of the acoustic differentiation of fear, anger, sadness, and joy. Thirty-one female subjects between the ages of 18 and 35 (encoders) were audio-recorded during an emotion-induction procedure and produced a total of 620 emotion-laden sentences. Twelve female judges (decoders), three for each of the four emotions, were assigned to rate the intensity of one emotion each. Their combined ratings were used to select 38 prototype samples per emotion. Past acoustic findings were replicated, and increased acoustic differentiation among the emotions was achieved. Multiple regression analysis suggested that some, although not all, of the acoustic variables were associated with decoders' ratings. Signal detection analysis gave some insight into this disparity. However, the analysis of the classic constellation of acoustic variables may not completely capture the acoustic features that influence decoders' ratings. Future analyses would likely benefit from the parallel assessment of respiration, phonation, and articulation. PMID- 10380662 TI - Real-time processing implications of enriched composition at the syntax-semantics interface. AB - This study reports results on the real-time consequences of aspectual coercion. We define aspectual coercion as a combinatorial semantic operation requiring computation over and above that provided by combining lexical items through expected syntactic processes. An experiment is described assessing whether or not parsing of a string requiring coercion--in addition to syntactic composition--is more computationally costly than parsing a syntactically transparent counterpart, a string that provides for an interpretable representation via syntactic composition alone. The prediction of a higher computational cost for this process is borne out by the results. PMID- 10380661 TI - Categorization of ambiguous sentences as a function of a changing prosodic parameter: a dynamical approach. AB - The present study investigates the dynamics of changes in interpretation of ambiguous sentences. The sentences used had alternative interpretations with different surface structures (bracketing). Continua were created by systematic manipulation of prosodic cues (relative foot duration), resulting in stimulus sentences that spanned the range between the two interpretations. Continua were presented to subjects who were requested to indicate as quickly as possible which meaning they perceived. The pattern of responses and response times revealed the presence of hysteresis. That is, when the values of prosodic parameters are congruent with both interpretations, the individual's recent history decides which meaning will be perceived. Thus, we can treat this categorization process as a transition from an initially stable meaning that loses stability with variations in prosody (our control parameter). The same underlying dynamics have been observed in studies of perception of syllables and aspects of visual perception. Apart from demonstrating the same characteristics of pattern formation at various levels of cognition, the study points to the usefulness of the dynamical approach in the investigation of language understanding. PMID- 10380663 TI - Adaptation of 2f1-2f2 distortion product otoacoustic emission in young-adult and old CBA and C57 mice. AB - The phenomenon of efferent-mediated adaptation of 2f1-f2 distortion product otoacoustic emission (DPOAE) was investigated in two strains (CBA/JNia and C57BL/6JNia) of mice of various ages using stimuli presented monaurally or binaurally. The present study demonstrated the existence of the DPOAE adaptation phenomenon in mice analogous to that previously reported in cats. The present data were fitted with one- or two-exponential functions. With a one-exponential fit in 2-month old mice, the adaptation magnitude ranged from 0 to 4 dB with the average value of 0.5 to 1.6 dB and the average time constant was 0.5 to 2.3 s. With a two-exponential fit, the shorter time constant was 0.3 to 1.7 s. The adaptation magnitude and time constant were similar between the monaural and binaural stimulations. We observed that there was a statistically significant decrease of adaptation magnitude in older CBA mice with age-related hearing loss when compared with young adult mice. The results from the young adult mice should be useful in future studies, e.g., a study of developmental changes in post-natal mice, or changes accompanying an alteration in the central auditory system arising from any etiology. We suggest that this phenomenon can be used as a tool for advancing basic knowledge of the auditory system and for assessing an impairment of the olivocochlear system, e.g., in aging. PMID- 10380664 TI - The "inverse problem" solved for a three-dimensional model of the cochlea. III. Brushing-up the solution method. AB - In two earlier papers [de Boer, J. Acoust. Soc. Am. 98, 896-903 and 904-910 (1995)] the inherent problems of the inverse-solution method in cochlear mechanics were described. The present paper shows results obtained with a more universal solution method. With the new method it is possible to construct a three-dimensional model of the cochlea producing a response that accurately simulates a measured mechanical basilar-membrane response. With earlier methods this could not be done. The inverse solution invariably yields that, with low stimulus levels, the model simulating a viable cochlea must be locally active. For the response of a dead animal a passive model is sufficient. Once more the inherent intricacies and problems of the inverse-solution method are discussed. Conservation of fluid volume leads to the concept of the "virtual stapes velocity." For best results, the input signal to the inverse-solution procedure should be acquired in the form of a "composite cross-correlation spectrum." Inverse analysis can, under certain circumstances, produce aberrant results. In this paper it is shown why the resulting impedance function is the most accurate in the region of the response peak. Therefore, it is unlikely that a passive model would exist of which the response simulates the data obtained from a healthy animal. PMID- 10380665 TI - Dolphin hearing: relative sensitivity as a function of point of application of a contact sound source in the jaw and head region. AB - The auditory input area of the dolphin head was investigated in an unrestrained animal trained to beach itself and to accept noninvasive electroencephalograph (EEG) electrodes for the recording of the auditory brain-stem response (ABR). The stimulus was a synthetic dolphin click, transmitted from a piezo-electric transducer and coupled to the skin via a small volume of water. The results conform with earlier experiments on acute preparations that show best auditory sensitivity at the middle of the lower jaw. Minimum latency was found at the rear of the lower jaw. A shaded receiver configuration for the dolphin ear is proposed. PMID- 10380666 TI - Intensity discrimination of Gaussian-windowed tones: indications for the shape of the auditory frequency-time window. AB - The just-noticeable difference in intensity jnd(I) was measured for 1-kHz tones with a Gaussian-shaped envelope as a function of their spectro-temporal shape. The stimuli, with constant energy and a constant product of bandwidth and duration, ranged from a long-duration narrow-band "tone" to a short-duration broadband "click." The jnd(I) was measured in three normal-hearing listeners at sensation levels of 0, 10, 20, and 30 dB in 35 dB(A) SPL pink noise. At intermediate sensation levels, jnd(I) depends on the spectro-temporal shape: at the extreme shapes (tones and clicks), intensity discrimination performance is best, whereas at intermediate shapes the jnd(I) is larger. Similar results are observed at a higher overall sound level, and at a higher carrier frequency. The maximum jnd(I) is observed for stimuli with an effective bandwidth of about 1/3 octave and an effective duration of 4 ms at 1 kHz (1 ms at 4 kHz). A generalized multiple-window model is proposed that assumes that the spectro-temporal domain is partitioned into "internal" auditory frequency-time windows. The model predicts that intensity discrimination thresholds depend upon the number of windows excited by a signal: jnd(I) is largest for stimuli covering one window. PMID- 10380667 TI - Speech intelligibility and localization in a multi-source environment. AB - Natural environments typically contain sound sources other than the source of interest that may interfere with the ability of listeners to extract information about the primary source. Studies of speech intelligibility and localization by normal-hearing listeners in the presence of competing speech are reported on in this work. One, two or three competing sentences [IEEE Trans. Audio Electroacoust. 17(3), 225-246 (1969)] were presented from various locations in the horizontal plane in several spatial configurations relative to a target sentence. Target and competing sentences were spoken by the same male talker and at the same level. All experiments were conducted both in an actual sound field and in a virtual sound field. In the virtual sound field, both binaural and monaural conditions were tested. In the speech intelligibility experiment, there were significant improvements in performance when the target and competing sentences were spatially separated. Performance was similar in the actual sound field and virtual sound-field binaural listening conditions for speech intelligibility. Although most of these improvements are evident monaurally when using the better ear, binaural listening was necessary for large improvements in some situations. In the localization experiment, target source identification was measured in a seven-alternative absolute identification paradigm with the same competing sentence configurations as for the speech study. Performance in the localization experiment was significantly better in the actual sound-field than in the virtual sound-field binaural listening conditions. Under binaural conditions, localization performance was very good, even in the presence of three competing sentences. Under monaural conditions, performance was much worse. For the localization experiment, there was no significant effect of the number or configuration of the competing sentences tested. For these experiments, the performance in the speech intelligibility experiment was not limited by localization ability. PMID- 10380668 TI - Acoustic pursuit of invisible moving targets by cats. AB - Head movements evoked by an invisible acoustic target were used as a metric to analyze localization of moving sources of sound in naive cats. The target was presented in the lateral sound field and moved along an arc at constant angular speeds. Head-movement trajectories were characterized by a large-magnitude orienting component that undershot the target, and a tracking component elicited by the target during acoustic pursuit. The tracking component was characterized by a succession of stepwise head movements that maintained a relatively close alignment of the median plane of the head with the moving acoustic target. PMID- 10380669 TI - Sound localization in noise in hearing-impaired listeners. AB - The present study assesses the ability of four listeners with high-frequency, bilateral symmetrical sensorineural hearing loss to localize and detect a broadband click train in the frontal-horizontal plane, in quiet and in the presence of a white noise. The speaker array and stimuli are identical to those described by Lorenzi et al. (in press). The results show that: (1) localization performance is only slightly poorer in hearing-impaired listeners than in normal hearing listeners when noise is at 0 deg azimuth, (2) localization performance begins to decrease at higher signal-to-noise ratios for hearing-impaired listeners than for normal-hearing listeners when noise is at +/- 90 deg azimuth, and (3) the performance of hearing-impaired listeners is less consistent when noise is at +/- 90 deg azimuth than at 0 deg azimuth. The effects of a high frequency hearing loss were also studied by measuring the ability of normal hearing listeners to localize the low-pass filtered version of the clicks. The data reproduce the effects of noise on three out of the four hearing-impaired listeners when noise is at 0 deg azimuth. They reproduce the effects of noise on only two out of the four hearing-impaired listeners when noise is at +/- 90 deg azimuth. The additional effects of a low-frequency hearing loss were investigated by attenuating the low-pass filtered clicks and the noise by 20 dB. The results show that attenuation does not strongly affect localization accuracy for normal hearing listeners. Measurements of the clicks' detectability indicate that the hearing-impaired listeners who show the poorest localization accuracy also show the poorest ability to detect the clicks. The inaudibility of high frequencies, "distortions," and reduced detectability of the signal are assumed to have caused the poorer-than-normal localization accuracy for hearing-impaired listeners. PMID- 10380670 TI - Temporal integration of loudness in listeners with hearing losses of primarily cochlear origin. AB - To investigate how hearing loss of primarily cochlear origin affects the loudness of brief tones, loudness matches between 5- and 200-ms tones were obtained as a function of level for 15 listeners with cochlear impairments and for seven age matched controls. Three frequencies, usually 0.5, 1, and 4 kHz, were tested in each listener using a two-interval, two--alternative forced--choice (2I, 2AFC) paradigm with a roving-level, up-down adaptive procedure. Results for the normal listeners generally were consistent with published data [e.g., Florentine et al., J. Acoust Soc. Am. 99, 1633-1644 (1996)]. The amount of temporal integration- defined as the level difference between equally loud short and long tones--varied nonmonotonically with level and was largest at moderate levels. No consistent effect of frequency was apparent. The impaired listeners varied widely, but most showed a clear effect of level on the amount of temporal integration. Overall, their results appear consistent with expectations based on knowledge of the general properties of their loudness-growth functions and the equal-loudness ratio hypothesis, which states that the loudness ratio between equal-SPL long and brief tones is the same at all SPLs. The impaired listeners' amounts of temporal integration at high SPLs often were larger than normal, although it was reduced near threshold. When evaluated at equal SLs, the amount of temporal integration well above threshold usually was in the low end of the normal range. Two listeners with abrupt high-frequency hearing losses (slopes > 50 dB/octave) showed larger-than-normal maximal amounts of temporal integration (40 to 50 dB). This finding is consistent with the shallow loudness functions predicted by our excitation-pattern model for impaired listeners [Florentine et al., in Modeling Sensorineural Hearing Loss, edited by W. Jesteadt (Erlbaum, Mahwah, NJ, 1997), pp. 187-198]. Loudness functions derived from impaired listeners' temporal integration functions indicate that restoration of loudness in listeners with cochlear hearing loss usually will require the same gain whether the sound is short or long. PMID- 10380671 TI - System identification of feedback in hearing aids. AB - The feedback problems of behind the ear (BTE), in the ear (ITE), and in the ear canal (ITEC) hearing aid categories have been investigated. All possible feedback paths (acoustical via vent, via tubing wall, mechanical, etc.) were converted to a single transfer function from the ear canal to the hearing aid microphone, here called the acoustic feedback equivalent (AFE). The attenuation of the AFE represents the maximum gain that can be used without the hearing aid starting to howl. Magnitude and phase responses of the AFE were identified on ten human subjects and on a Knowles ear manikin (KEMAR). The acoustic feedback via vent and leak between earmould and ear canal dominated the AFE. The transfer function from a reference point under the ear to the position of microphone of the different hearing aid categories was identified and used together with the AFE to calculate the maximum real ear aided gain (REAG) for the hearing aid categories. A model of the AFE, consisting of a fourth-order filter together with a delay, showed good agreement with the measured data. PMID- 10380672 TI - Missing-data model of vowel identification. AB - Vowel identity correlates well with the shape of the transfer function of the vocal tract, in particular the position of the first two or three formant peaks. However, in voiced speech the transfer function is sampled at multiples of the fundamental frequency (F0), and the short-term spectrum contains peaks at those frequencies, rather than at formants. It is not clear how the auditory system estimates the original spectral envelope from the vowel waveform. Cochlear excitation patterns, for example, resolve harmonics in the low-frequency region and their shape varies strongly with F0. The problem cannot be cured by smoothing: lag-domain components of the spectral envelope are aliased and cause F0-dependent distortion. The problem is severe at high F0's where the spectral envelope is severely undersampled. This paper treats vowel identification as a process of pattern recognition with missing data. Matching is restricted to available data, and missing data are ignored using an F0-dependent weighting function that emphasizes regions near harmonics. The model is presented in two versions: a frequency-domain version based on short-term spectra, or tonotopic excitation patterns, and a time-domain version based on autocorrelation functions. It accounts for the relative F0-independency observed in vowel identification. PMID- 10380673 TI - Identification of resynthesized /hVd/ utterances: effects of formant contour. AB - The purpose of this study was to examine the role of formant frequency movements in vowel recognition. Measurements of vowel duration, fundamental frequency, and formant contours were taken from a database of acoustic measurements of 1668 /hVd/ utterances spoken by 45 men, 48 women, and 46 children [Hillenbrand et al., J. Acoust. Soc. Am. 97, 3099-3111 (1995)]. A 300-utterance subset was selected from this database, representing equal numbers of 12 vowels and approximately equal numbers of tokens produced by men, women, and children. Listeners were asked to identify the original, naturally produced signals and two formant synthesized versions. One set of "original formant" (OF) synthetic signals was generated using the measured formant contours, and a second set of "flat formant" (FF) signals was synthesized with formant frequencies fixed at the values measured at the steadiest portion of the vowel. Results included: (a) the OF synthetic signals were identified with substantially greater accuracy than the FF signals; and (b) the naturally produced signals were identified with greater accuracy than the OF synthetic signals. Pattern recognition results showed that a simple approach to vowel specification based on duration, steady-state F0, and formant frequency measurements at 20% and 80% of vowel duration accounts for much but by no means all of the variation in listeners' labeling of the three types of stimuli. PMID- 10380674 TI - Preferred self-to-other ratios in choir singing. AB - Choir singers need to hear their own voice in an adequate self-to-other ratio (SOR) over the rest of the choir. Knowing singers' preferences for SOR could facilitate the design of stages and of choral formations. In an experiment to study the preferred SOR, subjects sang sustained vowels together with synthesized choir sounds, whose loudness tracked that of their own voice. They could control the SOR simply by changing their distance to the microphone. At the most comfortable location, the SOR was measured. Experimental factors included unison and four-part tasks, three vowels and two levels of phonation frequency. The same experiment was run four times, using sopranos, altos, tenors, and basses, with stimulus tones adapted for each category. The preferred self-to-other ratios were found to be similar to SORs measured previously in actual performance, if a little higher. Preferences were quite narrow, typically +/- 2 dB for each singer, but very different from singer to singer, with intrasubject means ranging from -1 to +15 dB. There was no significant difference between the unison and the four part tasks, although this might have been caused by systematic differences in the stimulus sounds. Some effects of phonation frequency and vowel were significant, but interdependent and difficult to interpret. The results and their relevance to live choir singing are discussed. PMID- 10380675 TI - The underwater audiogram of the West Indian manatee (Trichechus manatus). AB - The hearing thresholds of two adult manatees were measured using a forced-choice two alternative paradigm and an up/down staircase psychometric method. This is the first behavioral audiogram measured for any Sirenian, as well as the first underwater infrasonic psychometric test with a marine mammal. Auditory thresholds were obtained from 0.4 to 46 kHz, and detection thresholds of possible vibrotactile origin were measured at 0.015-0.2 kHz. The U-shaped audiogram demonstrates an upper limit of functional hearing at 46 kHz with peak frequency sensitivity at 16 and 18 kHz (50 dB re: 1 microPa). The range of best hearing is 6-20 kHz (approximately 9 dB down from maximum sensitivity). Sensitivity falls 20 dB per octave below 0.8 kHz and approximately 40 dB per octave above 26 kHz. The audiogram demonstrates a wider range of hearing and greater sensitivity than was suggested from evoked potential and anatomical studies. High frequency sensitivity may be an adaptation to shallow water, where the propagation of low frequency sound is limited by physical boundary effects. Hearing abilities of manatees and other marine mammals may have also been shaped by ambient and thermal noise curves in the sea. Inadequate hearing sensitivity at low frequencies may be a contributing factor to the manatees' inability to effectively detect boat noise and avoid collisions with boats. PMID- 10380676 TI - Human body odour, symmetry and attractiveness. AB - Several studies have found body and facial symmetry as well as attractiveness to be human mate choice criteria. These characteristics are presumed to signal developmental stability. Human body odour has been shown to influence female mate choice depending on the immune system, but the question of whether smell could signal general mate quality, as do other cues, was not addressed in previous studies. We compared ratings of body odour, attractiveness, and measurements of facial and body asymmetry of 16 male and 19 female subjects. Subjects wore a T shirt for three consecutive nights under controlled conditions. Opposite-sex raters judged the odour of the T-shirts and another group evaluated portraits of the subjects for attractiveness. We measured seven bilateral traits of the subject's body to assess body asymmetry. Facial asymmetry was examined by distance measurements of portrait photographs. The results showed a significant positive correlation between facial attractiveness and sexiness of body odour for female subjects. We found positive relationships between body odour and attractiveness and negative ones between smell and body asymmetry for males only if female odour raters were in the most fertile phase of their menstrual cycle. The outcomes are discussed in the light of different male and female reproductive strategies. PMID- 10380678 TI - Competition for neurotrophic factor in the development of nerve connections. AB - The development of nerve connections is thought to involve competition among axons for survival promoting factors, or neurotrophins, which are released by the cells that are innervated by the axons. Although the notion of competition is widely used within neurobiology, there is little understanding of the nature of the competitive process and the underlying mechanisms. We present a new theoretical model to analyse competition in the development of nerve connections. According to the model, the precise manner in which neurotrophins regulate the growth of axons, in particular the growth of the amount of neurotrophin receptor, determines what patterns of target innervation can develop. The regulation of neurotrophin receptors is also involved in the degeneration and regeneration of connections. Competition in our model can be influenced by factors dependent on and independent of neuronal electrical activity. Our results point to the need to measure directly the specific form of the regulation by neurotrophins of their receptors. PMID- 10380677 TI - Neuronal population activity and functional imaging. AB - Human functional brain imaging detects blood flow changes that are thought to reflect the activity of neuronal populations and, thus, the responses of neurons that carry behaviourally relevant information. Since this relationship is poorly understood, we explored the link between the activity of single neurons and their neuronal population. The functional imaging results were in good agreement with levels of population activation predicted from the known effects of sensory stimulation, learning and attention on single cortical neurons. However, the nature of the relationship between population activation and single neuron firing was very surprising. Population activation was strongly influenced by those neurons firing at low rates and so was very sensitive to the baseline or 'spontaneous' firing rate. When neural representations were sparse and neurons were tuned to several stimulus dimensions, population activation was hardly influenced by the few neurons whose firing was most strongly modulated by the task or stimulus. Measures of population activation could miss changes in information processing given simultaneous changes in neurons' baseline firing, response modulation or tuning width. Factors that can modulate baseline firing, such as attention, may have a particularly large influence on population activation. The results have implications for the interpretation of functional imaging signals and for cross-calibration between different methods for measuring neuronal activity. PMID- 10380679 TI - Evolutionary affinities of the enigmatic saola (Pseudoryx nghetinhensis) in the context of the molecular phylogeny of Bovidae. AB - To elucidate the systematic status of the enigmatic saola (Pseudoryx nghetinhensis), a new bovid genus recently discovered in Vietnam, and to investigate phylogenetic relationships within the family Bovidae, four distinct DNA markers were sequenced. Complete mitochondrial cytochrome b (1143 bp) and 12S rRNA (956 bp) genes and non-coding regions from the nuclear genes for aromatase cytochrome P-450 (199 bp) and lactoferrin (338 bp) have been compared for 25 bovid species and three Cervidae and Antilocapridae outgroups. Independent and/or combined analyses of the four nucleotide matrices through maximum parsimony and maximum-likelihood methods indicated that Bovidae consists of two major lineages, i.e. Bovinac which contains the tribes Bovini, Boselaphini and Tragelaphini, and Antilopinae which encompasses all other bovids. Within Bovinae, the tribe Bovini is divided into buffalo Bovini (Bubalus and Syncerus) and cattle Bovini (Bos and Bison) and Tragelaphini are possibly related to Boselaphini. Pseudoryx is shown to be (i) robustly nested within Bovinae; (ii) strongly associated with Bovini; and (iii) tentatively sharing a sister-group relationship with cattle Bovini. Within Antilopinae, three robust clades are in evidence: (i) Hippotragus and Damaliscus are linked to Ovis; (ii) Aepyceros joins Neotragus; and (iii) Cephalophus clusters with Oreotragus. PMID- 10380680 TI - DNA analyses support the hypothesis that infanticide is adaptive in langur monkeys. AB - Although the killing of dependent infants by adult males is a widespread phenomenon among primates, its causes and consequences still remain hotly debated. According to the sexual selection hypothesis, infanticidal males will gain a reproductive advantage provided that only unrelated infants are killed and that the males increase their chances of siring the next infants. Alternatively, the social pathology hypothesis interprets infanticide as a result of crowded living conditions and, thus, as not providing any advantage. Based on DNA analyses of wild Hanuman langurs (Presbytis entellus) we present the first evidence that male attackers were not related to their infant victims. Furthermore, in all cases the presumed killers were the likely fathers of the subsequent infants. Our data, therefore, strongly support the sexual selection hypothesis interpreting infanticide as an evolved, adaptive male reproductive tactic. PMID- 10380681 TI - Human parieto-occipital visual cortex: lack of retinotopy and foveal magnification. AB - We studied visual representation in the parietal cortex by recording whole-scalp neuromagnetic responses to luminance stimuli of varying eccentricities. The stimuli were semicircles (5.5 degrees in radius) presented at horizontal eccentricities from 0 degree to 16 degrees, separately in the right and left hemifields. All stimuli evoked responses in the contralateral occipital and medial parietal areas. The waveforms and distributions of the occipital responses varied with stimulus side (left, right) and eccentricity, whereas the parietal responses were remarkably similar to all stimuli. The equivalent sources of the parietal signals clustered within 1 cm3 in the medial parieto-occipital sulcus and did not differ significantly between the stimuli. The strength of the parietal activation remained practically constant with increasing stimulus eccentricity, suggesting that the visual areas in the parieto-occipital sulcus lack the enhanced foveal representation typical of most other visual areas. This result strengthens our previous suggestion that the medial parieto-occipital sulcus is the human homologue of the monkey V6 complex, characterized by, for example, lack of retinotopy and the absence of relative foveal magnification. PMID- 10380682 TI - Spatial heterogeneity and function of voltage- and ligand-gated ion channels in retinal amacrine neurons. AB - The spatial distribution of ion channels within amacrine cells of the tiger salamander retina was studied using patch recording in the retinal slice preparation. By focally puffing kainate, GABA and glycine at amacrine cell processes in the inner plexiform layer, it was determined that the cell's glutamate receptors were located in a confined region of the processes near the soma, while glycine and GABA receptors were located throughout the processes. Likewise, similar techniques in conjunction with voltage steps demonstrated that voltage-gated sodium channels were located throughout the cell and were shown to generate sodium-dependent spikes, while only the processes contained voltage gated calcium channels. These results suggest that this form of transient amacrine cell collects its excitatory synaptic inputs in a region confined to a central annular region near the soma, that the signal is actively propagated throughout its processes by voltage-gated sodium channels and that calcium dependent neurotransmitter release of glycine from this neuron can occur throughout its processes. Thus, excitatory signals are collected in the processes near the soma, inhibitory signals throughout the processes and excitation is probably propagated throughout the processes of the amacrine cell. PMID- 10380683 TI - T-cell induced pathogenesis in HIV: bystander effects and latent infection. AB - The progress of HIV is accompanied by the infection and decline of the population of CD4+ cells. This reduction in cells results from both cytolytic influences of the virus and virus-specific cytotoxic T-cell (CTL) responses. We seek to characterize the extent of CD4+ reduction caused by HIV-specific CTLs at equilibrium. Here we show that intermediate levels of cytotoxic killing of infected cells can be inferior to both strong and weak or absent immune responses. We further show that the deleterious effects of the CTL response are made worse by a slow immune response. Bystander effects in which uninfected cells are thought to be eliminated by non-specific CTL activation lead to small or negligible reductions in uninfected CD4+ cells. Latently infected cells containing pro-viral DNA and which become activated at a constant rate ensure that the immune response is more effective for a larger range of CTL activities and reduces T-cell associated pathology. PMID- 10380684 TI - Epidermal diseases in bottlenose dolphins: impacts of natural and anthropogenic factors. AB - Experimental studies have highlighted the potential influence of contaminants on marine mammal immune function and anthropogenic contaminants are commonly believed to influence the development of diseases observed in the wild. However, estimates of the impact of contaminants on wild populations are constrained by uncertainty over natural variation in disease patterns under different environmental conditions. We used photographic techniques to compare levels of epidermal disease in ten coastal populations of bottlenose dolphins (Tursiops truncatus) exposed to a wide range of natural and anthropogenic conditions. Epidermal lesions were common in all populations (affecting > 60% of individuals), but both the prevalence and severity of 15 lesion categories varied between populations. No relationships were found between epidermal disease and contaminant levels across the four populations for which toxicological data were available. In contrast, there were highly significant linear relationships with oceanographic variables. In particular, populations from areas of low water temperature and low salinity exhibited higher lesion prevalence and severity. Such conditions may impact on epidermal integrity or produce more general physiological stress, potentially making animals more vulnerable to natural infections or anthropogenic factors. These results show that variations in natural environmental factors must be accounted for when investigating the importance of anthropogenic impacts on disease in wild marine mammals. PMID- 10380685 TI - The basic reproduction number for scrapie. AB - The basic reproduction number R0 provides a quantitative assessment of the ability of an infectious agent to invade a susceptible host population. A mathematical expression for R0 is derived based on a recently developed model for the spread of scrapie through a flock of sheep. The model incorporates sheep demography, a long and variable incubation period, genetic variation in susceptibility to scrapie, and horizontal and vertical routes of transmission. The sensitivity of R0 to a range of epidemiologically important parameters is assessed and the effects of genetic variation in susceptibility are examined. A reduction in the frequency of the susceptibility allele reduces R0 most effectively when the allele is recessive, whereas inbreeding may increase R0 when the allele is recessive, increasing the chance of an outbreak. Using this formulation, R0 is calculated for an outbreak of scrapie in a flock of Cheviot sheep. PMID- 10380686 TI - Ambulatory blood pressure monitoring: a proposal for new Canadian guidelines. Division of Cardiology, Toronto, Ontario. AB - OBJECTIVE: To provide background information on the scientific basis for new Canadian guidelines on ambulatory blood pressure monitoring and the process for developing recommendations for clinical practice. DATA SOURCES: A comprehensive review of the literature on ambulatory blood pressure monitoring was conducted using a computerized literature search and a bibliography of key scientific articles. STUDY SELECTION AND DATA EXTRACTION: Studies of the highest scientific quality were used to support recommendations based on established rules for grading evidence. DATA SYNTHESIS: A series of recommendations for the use of ambulatory blood pressure monitoring in routine clinical practice directed towards both untreated patients with a diagnosis of possible hypertension and treated patients with a suspected 'white coat' component to their office readings have been proposed for consideration by the Canadian Hypertension Society. CONCLUSIONS: There are now sufficient data available to support the use of ambulatory blood pressure monitoring in accordance with specific clinical practice guidelines. PMID- 10380687 TI - Intermittent blood pressure control: potential consequences for outcome. AB - Although both blood pressure (BP) and left ventricular (LV) mass at initial evaluation predict future cardiovascular risk, the actual BP and LV mass achieved over years of treatment more clearly relate to cardiovascular event rates. Intermittent compliance or noncompliance is the major reason for uncontrolled hypertension and presumably persistent LV hypertrophy. In general, drugs with rapid onset and short duration of action are not desirable because this profile may lead to large variations in BP lowering effect during actual drug intake and rapid disappearance of the antihypertensive effect with missed doses. In addition, intermittent compliance per se introduces the potential for adverse events. For drugs requiring several dose-titrations (e.g., alpha1-blockers), restarting at full doses may lead to excessive drug action and symptomatic hypotension. For other drugs (e.g., short acting beta-blockers or clonidine-like drugs), sudden discontinuation with intermittent compliance may lead to rebound enhanced sympathetic responsiveness after one to two days, resulting not only in side effects, but also in adverse events, particularly in patients with (silent) coronary artery disease. The rapid onset, short acting dihydropyridines cause intermittent BP control at each dosing, particularly at higher doses. This intermittent control of BP is even more apparent at dosing intervals that are long relative to the duration of action. Thus, sympathetic activation and potential for adverse events can be anticipated at each dosing unless these drugs are being taken frequently at relatively low doses. For diuretics, angiotensin converting enzyme inhibitors and angiotensin I receptor blockers, no adverse effects have been identified with intermittent compliance. Intermittent BP control is, in general, not an appropriate approach to the management of hypertension and introduces additional risks depending on the type of antihypertensive drug. In contrast, drugs with slow onset and long duration of action provide a more consistent effect during actual drug intake and a more persistent effect during short periods of noncompliance. PMID- 10380688 TI - Angiotensin-converting enzyme inhibitors and end-organ damage in heart failure. AB - Angiotensin-converting enzyme (ACE) inhibitors play an important role in protecting various organs in patients with congestive heart failure. The mechanisms of action of ACE inhibitors in congestive heart failure are multiple and may involve important effects on endothelial function in addition to the well known hemodynamic and neurohormonal effects. Skeletal muscle fatigue can play an important role in the pathophysiology of congestive heart failure, and only aggressive treatment with ACE inhibitors and exercise training can improve exercise tolerance and reduce the risk of further deterioration of cardiac function. ACE inhibitors should be introduced with caution in patients with severe heart failure and hypotension to prevent renal dysfunction. Unlike some other ACE inhibitors, perindopril seems to cause no first dose hypotension. It also appears to have a neutral effect on renal function in patients with severe congestive heart failure. PMID- 10380689 TI - Surgery of the aortic valve. PMID- 10380690 TI - Characterization of the MICA polymorphism by sequence-specific oligonucleotide probing. AB - A large number of diseases occur in association with specific HLA-B or -C alleles. Recently a new gene, termed major histocompatibility complex class I chain-related gene A (MICA), has been identified in close proximity to HLA-B. The function of this gene is still unknown, but, it is structurally related to HLA class I genes, is polymorphic, and is potentially associated with several diseases. Some DNA-based techniques have previously been described to type for MICA including sequencing and single-strand conformational polymorphism. In this paper we describe the application of sequence-specific oligonucleotide probe based typing for the analysis of the MICA gene. We used a set of 30 oligonucleotide probes to screen for the polymorphisms in exons 2, 3, and 4, which account for the 16 known alleles. We report here the typing results of MICA for 103 B-cell lines that have been well characterized for HLA and describe the linkage disequilibrium between MICA and HLA-B. Unequivocal MICA typing was achieved for 85 of the 103 cells tested, 6 cells gave ambiguous MICA types, and a further 12 cells showed patterns consistent with them expressing at least one new MICA allele. PMID- 10380691 TI - A divergent non-classical class I gene conserved in salmonids. AB - Complementary DNA for two class I genes of the rainbow trout, Oncorhynchus mykiss, were characterized. MhcOnmy-UBA*01 is similar to Onmy-UAC32 and the classical major histocompatibility complex class I genes of other fish species, whereas Onmy-UAA*01 is divergent from all class I genes so far characterized. Onmy-UAA*01 is expressed at lower levels than Onmy-UBA*01. Although Onmy-UAA*01 exhibits restriction fragment length polymorphism on Southern blotting, the encoded protein is highly conserved. Two allotypes, which differ only by substitution at amino acid position 223 of the alpha 3 domain, have been defined. Onmy-UAA*01 has an exon-intron organization like other class I genes and contains a Tc1-like transposon element in intron III. Orthologues of Onmy-UAA*01 have been characterized in four other species of salmonid. Between four species of Oncorhynchus, UAA*01 proteins differ by only 2-6 amino acids, whereas comparison of Oncorhynchus with Salmo trutta (brown trout) reveals 14-16 amino acid differences. The Onmy-UAA*01 gene has properties indicative of a particularly divergent non-classical class I gene. PMID- 10380692 TI - Ancestral and recombinant 16-locus HLA haplotypes in the Hutterites. AB - Prior studies in the Schmiedeleut Hutterites of South Dakota have demonstrated associations between human leukocyte antigen (HLA) haplotype matching and fetal loss (Ober et al. 1992) and mate preferences (Ober et al. 1997), as well as deficiencies of homozygotes for HLA haplotypes (Kostyu et al. 1993). These studies were based on the serologically-defined five-locus HLA-A, -C, -B, -DR, DQ haplotype. To further elucidate the effects of specific major histocompatibility (MHC) loci or regions on fetal loss and mate choice, we genotyped a sample of Hutterites for 14 MHC loci by DNA or biochemical methods. Typing for additional loci in the HLA-A to HLA-DPB1 region increased the number of recognized Hutterite MHC haplotypes to 67, and further localized the site of crossover in 9 of 15 recombinant haplotypes. Hutterite MHC haplotype sequences are similar to those observed in outbred Caucasians, suggesting that the influence of HLA haplotypes on fetal loss and mating structure may be general. PMID- 10380693 TI - The mouse p52 subunit of the transcription/DNA repair factor TFIIH is located in the class III region of the H2 complex: cloning and sequence polymorphism. AB - Loci controlling susceptibility to a number of diseases, including cortisone induced cleft palate, experimental allergic orchitis, and chemically-induced transplacental lung tumors have been mapped to a 27 kilobase (kb) region within the class III region of the mouse major histocompatibility complex (H2). This region, contains three genes G7e, which resembles a viral envelope gene, Bat6 (G7a), which encodes a valyl-tRNA synthetase, and G7c, which has no known function. We cloned a set of overlapping cosmid clones containing 115 kb of DNA surrounding Bat6. Exon trapping has identified a new gene located telomeric of Bat6. Northern blot analysis detected a transcript of 1.7 kb with highest expression in the testis. DNA sequence analysis identified this gene as the mouse homologue of the human gene encoding the p52 subunit of the TFIIH transcription/DNA repair factor. Nucleotide sequence identity was 91% between mouse and human, and the protein sequence was 98% identical. Sequence analysis of p52 cDNA from congenic mouse strains detected an amino acid polymorphism at position 209, which results in the substitution of a threonine in the H2b haplotype to a methionine in the H2a,d haplotypes. PMID- 10380694 TI - A TaqI polymorphism in the chicken interferon regulatory factor 1 (IRF1) gene provides evidence that chicken linkage group E48C28W13 represents one of the microchromosomes. PMID- 10380695 TI - Mapping of a candidate region for susceptibility to inclusion body myositis in the human major histocompatibility complex. AB - Inclusion body myositis (IBM) is a form of idiopathic inflammatory myopathy of unknown aetiology. A strong association with HLA class II (HLA-DR3) suggested a role for genes in the human major histocompatibility complex (MHC) in the predisposition to this disease. In this study, we have taken advantage of the ancestral haplotype (AH) concept and historical recombinations to map for a possible susceptibility gene(s) in the MHC. We performed detailed typing of three MHC-related HSP70 genes and defined allelic combinations in the context of MHC AH. We also modified existing methods to give a simple and accurate method for typing two TNF microsatellites. Using the HSP70 and TNF markers and HLA-DR, -B, and C4 typing of our patients with IBM, we defined a potential site for the MHC associated susceptibility gene(s) in the region between HLA-DR and C4. PMID- 10380696 TI - Comparison of allele O sequences of the human and non-human primate ABO system. AB - Like humans, non-human primates express the antigens A and B of the ABO histoblood group system. In chimpanzees, only A and O types are found, while the types A, B, AB, and O are found in macaques. The sequences of exons 6 and 7 of two chimpanzee O alleles (Odel and O(x), two macaque species O alleles (rhesus monkey and crab-eating macaque), and sequences of exon 7 of two major chimpanzee A alleles (A1ch and A2ch) were established. The sequences of cDNAs corresponding to the chimpanzee and rhesus monkey O alleles were characterized from exon 1 to 7 and from exon 4 to 7, respectively. A comparison of our results with ABO gene sequences already published by others demonstrates that human and non-human primate O alleles are species-specific and result from independent silencing mutations. These observations reinforce the hypothesis that the maintenance of the ABO gene polymorphism in primates reflects convergent evolution more than transpecies inheritance of ancestor alleles. PMID- 10380697 TI - IL1B gene polymorphisms influence the course and severity of inflammatory bowel disease. AB - There is evidence of a disbalance in the inflammatory regulation of patients with inflammatory bowel diseases (IBD). Interleukin-1 beta plays an important role in the pro-inflammatory response. Our aim was to study the influence which IL1B gene polymorphisms may have on the severity and course of these diseases. Ninety-six patients with ulcerative colitis (UC), 98 patients with Crohn's disease (CD), and 132 ethnically matched healty individuals (HC) were typed for the polymorphic sites in the promoter region (position -511) and in exon 5 (position +3953) of the IL1B gene, using polymerase chain reaction (PCR)-based methods. In the CD group a significant association (P = 0.009) was found in this pair of genes. Homozygotes for allele 1 at position +3953 were more often present (69% vs 31%) in the subgroup of patients carrying at least one copy of allele 2 at position 511. This association was significant in patients with non-perforating disease (P = 0.002), but was not present in patients with perforating-fistulizing disease. The distribution of both allelic pairs in the non-fistulizing group proved to be significantly different from HC (P < 0.05), UC (P < 0.03), and the fistulizing group (P < 0.05). There was a similar association in non-operated patients (P = 0.024), whereas no such association was found in surgically treated patients. Among carriers of allele 2 at position -511, UC patients with more severe bleeding symptoms (P = 0.006) were less frequently found. These results suggest that IL1B gene polymorphisms participate in determining the course and severity of inflammatory bowel disease and contribute to explain the heterogeneity of these diseases. PMID- 10380698 TI - Biased T-cell receptor usage is associated with allelic variation in the MHC class II peptide binding groove. AB - A comprehensive analysis was carried out of the tri-molecular complex of peptide, major histocompatibility class II molecule, and T-cell receptor (TcR) involved in the recognition of the promiscuous HA (306-318) peptide, restricted by one of two closely related HLA-DR alleles, HLA-DRB1*0101 and HLA-DRB1*0103. These two DR molecules differ by only three amino acids at positions 67, 70, and 71, in the third variable region of the DRB1 chain. None of the HA (306-318)-specific T-cell clones restricted by these two DR molecules tolerated amino acid substitution at the peptide-binding position 71, despite the fact that the substitution did not interfere with peptide binding. The majority of the DRB1*0103-restricted clones tolerated substitution of the amino acid at the TcR-contacting position 70, while the DRB1*0101-restricted T cells did not. Based usage of TRVA and TRVB segments was observed for the DRB1*0103-restricted clones; in contrast, apparently random usage was seen in the DRB1*0101-restricted T cells. Finally, limiting dilution analysis revealed a lower frequency of T cells reactive with the HA peptide in a DRB1*0103 compared with a DRB1*0101 individual. Taken together these data suggest that biased TcR gene usage may reflect a relatively low precursor frequency of T cells, and the need for clonal expansion of a limited set of high avidity T cells. PMID- 10380699 TI - Cloning, sequencing, and characterization of dog interleukin-18. AB - Interleukin-18, originally termed interferon-gamma inducing factor, is a recently described cytokine which is intimately involved in the generation of the immune response. The human and mouse sequences have been reported and studies have demonstrated the potent biological functions of this protein including the induction of interferon-gamma and the enhancement of NK cytotoxicity. This paper describes the cloning, sequencing, and characterization of the dog homologue of this gene. The coding sequence for dog IL-18 was obtained using reverse transcription-polymerase chain reaction from mRNA harvested from PMA-stimulated alveolar macrophages. Sequence analysis of the dog gene has demonstrated an open reading frame of 582 base pairs coding for a 193 amino acid precursor protein. The dog coding sequence shares 84% and 74% similarity to the human and mouse equivalents, respectively, at the nucleotide level. Based upon sequence analysis, we propose that this new gene is the dog equivalent of human IL-18. PMID- 10380700 TI - New family of Mhc class II A genes identified from cDNA sequences in the cichlid fish Aulonocara hansbaenschi. PMID- 10380701 TI - A radiation hybrid map of complement factor H and factor H-related genes. PMID- 10380702 TI - Genomic organization and chromosomal localization of the mouse lipopolysaccharide binding protein gene. PMID- 10380703 TI - MICA haplotypic diversity. PMID- 10380704 TI - Sequencing-based typing of MICA reveals 33 alleles: a study on linkage with classical HLA genes. PMID- 10380705 TI - The peptide-binding motif of HLA-A*0217. PMID- 10380706 TI - The HLA-A*6601 peptide motif: prediction by pocket structure and verification by peptide analysis. PMID- 10380707 TI - New variations of human SHP-1. PMID- 10380708 TI - Molecular cloning of cDNAs encoding dog high-affinity IgE receptor alpha, beta, and gamma chains. PMID- 10380709 TI - Cloning and sequence comparison of sheep CD28 and CTLA-4. PMID- 10380710 TI - Additional upstream coding sequences of MAGE-11. PMID- 10380711 TI - Characterization of the human myeloid IgA Fc receptor I (CD89) gene in a cosmid clone. PMID- 10380712 TI - A new mouse lymphocyte antigen encoded by a gene in the Nk complex. PMID- 10380713 TI - Treatment of schizophrenia 1999. The expert consensus guideline series. PMID- 10380714 TI - 5th International Symposium on Myelodysplastic Syndromes. Prague, Czech Republic, 21-24 April 1999. Abstracts. PMID- 10380715 TI - 20th Annual meeting of the Society for Clinical Trials. Anaheim, California, USA. May 2-5, 1999. Abstracts. PMID- 10380716 TI - Breast cancer: a guide for fellows. PMID- 10380717 TI - 14th International Conference on Oral and Maxillofacial Surgery. Washington DC, USA. 24-29 April 1999. Abstracts. PMID- 10380718 TI - 20th Annual scientific sessions of the North American Society of Pacing and Electrophysiology (NASPE). Toronto, Ontario, Canada. May 12-15, 1999. Abstracts. PMID- 10380719 TI - German Physiological Society 78th annual meeting. Bonn, Germany, 14-17 March 1999. Abstracts. PMID- 10380720 TI - 38th Annual meeting of the Society of Toxicology. New Orleans, Louisiana, USA. March 14-18, 1999. Abstracts. PMID- 10380721 TI - Movement and muscle activity pattern in wheelchair ambulation by persons with para-and tetraplegia. AB - The patterns of movement and muscle activation in wheelchair ambulation have been studied in two groups: subjects with paraplegia (n = 4) and tetraplegia (n = 3). All subjects were physically active and experienced wheelchair users. The tests were done in the subjects' own wheelchairs and under free-wheeling conditions. The tasks studied were: self-chosen normal velocity, maximal velocity and maximally accelerated start. Muscle activation was registered by surface electromyography performed on several arm and shoulder muscles. The movement pattern was studied by goniometry of the shoulder and elbow joints, as well as by observing video recordings. Speed and arm cycle frequency were also recorded. The movement pattern was divided into three phases: pull, push and recovery. Relatively concordant muscle activation patterns were noted within the groups, whereas differences were noted between the groups with regard to muscle activation, length of the pull and push phases and the velocity-dependent adaptation. The subjects with tetraplegia were more dependent on the pull phase. The self-chosen normal and maximal speeds of the subjects with tetraplegia were approximately half those of the subjects with paraplegia. Three different types of recovery movements were noted as well as a velocity-dependent adaptation. Major trunk movements during the rim phase were only noted at the maximally accelerated start. In conclusion, the results point to both similarities and differences in the movement pattern and muscle activation in individuals with para- and tetraplegia under different ambulation conditions. The differences are of such a magnitude that they are important enough to consider when teaching wheelchair techniques and developing rehabilitation programmes for different groups of patients with spinal cord injuries. PMID- 10380722 TI - Restoring normal gait after limb salvage procedures in malignant bone tumours of the knee. AB - Eleven patients exhibiting decreased strength of knee extension following wide resection and prosthetic reconstruction for malignant bone tumors of the knee performed gait exercises with compensatory muscle training. Two patients whose knee extension strength was assessed as manual muscle test (MMT) grade 4 were able to develop a gait with double knee action and to maneuver stairs, step-by step, due to compensation by the gluteus maximus, biceps femoris, and gastrocnemius muscles. Four patients whose knee extension strength was less than MMT grade 4, and whose ankle dorsal and plantar flexion was MMT grade 4 or higher, acquired the ability to go up and down stairs step-by-step, although their gait pattern was a knee-extended gait. Electromyographic studies demonstrated continuous discharges of the gluteus maximus, biceps femoris, and gastrocnemius muscles during the stance phase as compensation for decreased strength in knee extension. PMID- 10380723 TI - A cross-validation of the comprehensive assessment of activities of daily living after stroke. AB - This study aimed to determine whether the Frenchay Activities Index and the Barthel Index assess different factors in stroke patients who survive for more than one year. The Frenchay Activities Index and the Barthel Index were administered via telephone interview. One hundred and twenty-four patients from the community participated in the study. All items of the Barthel Index and the Frenchay Activities Index, except reading books, were included in a factor analysis to determine the underlying constructs of the items. Four factors were found. One factor comprised all items from the Barthel Index and one item from the Frenchay Activities Index. The rest of the Frenchay Activities Index items loaded on three other factors. The combined scores, using simple transformation, had satisfactory distributions. The results support the hypothesis that the Frenchay Activities Index and the Barthel Index assess different factors in stroke patients who survive for more than one year. The Barthel Index score and the Frenchay Activities Index score could be combined to assess the entire range of activities of daily living functions in stroke. PMID- 10380724 TI - Pressure pain thresholds in different tissues in one body region. The influence of skin sensitivity in pressure algometry. AB - This study aimed at determining whether there are differences in pressure pain sensitivity in different tissues in the same body region when systematically assessed, before and after skin hypoesthesia. Pressure pain thresholds (PPTs) were assessed bilaterally in 15 healthy females at the bony part of the epicondylus lateralis humeri, at the belly of m. extensor carpi ulnaris and at m. brachioradialis where the superficial radial nerve branches pass underneath ("muscle/nerve" site). Following a double blind design, a local anaesthetic cream (EMLA) or a control cream was applied to the skin and PPTs were reassessed. The PPT was significantly (p < 0.001) lower at the "muscle/nerve" site than at the bony and "pure" muscle sites. The PPTs over the bony and "pure" muscle sites did not differ. There was no significant difference when PPTs were compared before and after application of EMLA cream. However, PPTs after control cream were lower (p < 0.001) over all examined areas than those obtained prior to cream application. Thus, EMLA cream increased PPTs compared to control sites in all examined areas (p < 0.001). Under the given circumstances, skin pressure pain sensitivity was demonstrated to influence the PPT. PMID- 10380725 TI - The American Paediatric Evaluation of Disability Inventory (PEDI). Applicability of PEDI in Sweden for children aged 2.0-6.9 years. AB - The American Paediatric Evaluation of Disability Inventory (PEDI) is a new instrument for evaluating functional performance in disabled children aged 6 months to 7.5 years. It was developed to determine a child's functional capacity and performance in three domains, self-care, mobility and social function, as reflected in scores on three scales: (i) functional skills (current capability in specific tasks), (ii) caregiver assistance (i.e. provided to facilitate the child's performance), and (iii) modifications (i.e. environmental or technical modifications needed to facilitate the child's function). The present study was designed to compare results obtained using the PEDI in a Swedish sample with the American normative data, and to analyse the content and relevance of PEDI items for use in Sweden. The PEDI was administered as a questionnaire in structured interview form to the parents of 52 non-disabled Swedish children aged 2.0-6.9 years, divided into ten age groups. Correlation analysis (Pearson's r) showed scores for the Swedish sample to manifest strong correlation with the respective American normative data, both for the functional skills (r = 0.90-0.98) and caregiver assistance (r = 0.93-0.99) scales, respectively. Scores for the modification scale were not compared. Thus, the results suggest the American normative data to be appropriate for reference purposes in Sweden. PMID- 10380726 TI - Effects of organized aerobic group training in elderly patients discharged after an acute coronary syndrome. A randomized controlled study. AB - The aim of this study was to compare the physiological effects of an individually adjusted outpatient group training programme to the standardized recommendations of walking in elderly patients (>65 years) discharged after an acute coronary episode. In all, 101 patients, 20 women and 81 men, aged 65-84 (mean 71) years, were randomized either to a supervised outpatient group training programme during three months (n = 50) or to a control group (n = 51). Exercise tolerance increased from 104 watts to 122 watts (p < 0.001) in the training group and from 102 watts to 105 watts (n.s.) in the control group. Self-estimated level of physical activity was higher in the patients in the training group than in the control group (p < 0.001), as was graded well-being (p < 0.05). Organized aerobic group training can easily be performed in elderly patients after acute coronary syndrome, with results of improved exercise tolerance and a higher self graded well-being. PMID- 10380727 TI - Effects of an exercise programme on organizational/psychosocial and physical work conditions, and psychosomatic symptoms. AB - The aim of this study was to evaluate the effect of a weekly exercise programme among nursing staff on organizational/psychosocial and physical work conditions, and psychosomatic symptoms. Out of 106 nurses and nursing aides from four geriatric wards who were invited to participate in a cross-over study, 86 accepted. For the exercise periods the staff were invited to participate in an exercise programme twice a week for 8 weeks during work time. Fifty subjects participated > or = 8 times regularly during the exercise periods (participants). During the control periods, 78 subjects attended without intervention. The effect was followed-up with questionnaires before and after the intervention periods. The exercise programme did not affect perceived organizational/psychosocial or physical work conditions, with one exception. A higher change for the worse was seen in the factor "work planning" during the exercise periods compared with during the control periods. The result suggests that the organization of the training is important in order not to add extra stress. PMID- 10380728 TI - Reduction of visuo-spatial neglect with vestibular galvanic stimulation. AB - The purpose of the present investigation was to determine the effect of galvanic vestibular stimulation on visuo-spatial neglect without inducing nystagmus and associated discomfort. Fourteen patients with right-hemisphere stroke with neglect were assessed with two visuo-motor tasks ("Line crossing" and "Star cancellation") on three occasions. Seven of the subjects received galvanic vestibular stimulation during the second condition (Experiment 1), whereas the other seven received stimulation during the third assessment (Experiment 2). Between-group comparisons of stimulation effects were performed by analyzing change on visuo-spatial neglect from the first to the second condition in the two experimental groups. A significantly larger effect was demonstrated on the "Line crossing" task in Experiment 1. This finding suggests a stimulation effect beyond practice/spontaneous recovery, and may provide new possibilities in rehabilitation research because the stimulation can be given without discomfort. PMID- 10380729 TI - Comments on "Measures of individual and group changes in ordered categorical data". PMID- 10380730 TI - Source and role of endolymph macromolecules. AB - Evaluation of some 200 endolymph proteins indicates that they are predominantly derived from plasma. However, the profile of endolymph proteins is remarkably similar to that of perilymph and entirely different from that of plasma. This supports the current consensus that perilymph rather than plasma is the (direct) source of endolymph. Although the levels of total protein of endolymph is extremely low, a few plasma-derived proteins, such as apolipoproteins J and D, are selectively enriched, conceivably for protection of cell membranes bounding the endolymphatic space. A small number of endolymph proteins, mostly glycosylated ones, are continually secreted into the endolymph by specialized epithelial cells, primarily for the maintenance of the structural and functional integrity of the extracellular superstructures comprising tectorial membrane, otoconial complex (membrane) and cupula. These complex macromolecules cannot be eliminated in the periphery of the compartment, but are transported to the endolymphatic sac for elimination. Impaired clearance of these negatively charged macromolecules by a dysfunctional endolymphatic sac will contribute to the chemical imbalance of endolymph which accompanies long-standing endolymphatic hydrops, and may be one of the reasons for the observed loss of function. PMID- 10380731 TI - Meniere's disease and endolymphatic hydrops without Meniere's symptoms: temporal bone histopathology. AB - We studied temporal bone histopathology in 21 ears with Meniere's disease and 24 ears with endolymphatic hydrops without Meniere's symptoms and compared the findings to those in 10 ears with presbycusis and 11 ears with normal hearing. Normal hearing ears showed less degeneration of cochlear structures than the other ears. In ears with endolymphatic hydrops without Meniere's symptoms, the degeneration of spiral ligament, hair cells, dendrites (peripheral processes) and apical spiral ganglion cells was more severe than in the other three groups. In ears with Meniere's disease and endolymphatic hydrops without Meniere's symptoms, the hair cells and dendrites were more affected than ganglion cells and there was no correlation between hair cell and ganglion cell degeneration. These findings suggest that a permanent threshold shift in late stage endolymphatic hydrops is not related to ganglion cell loss but rather to degeneration of sensory elements. PMID- 10380732 TI - Smoking as a risk factor in sensory neural hearing loss among workers exposed to occupational noise. AB - The effect of smoking on hearing was investigated among 199 professional forest workers and 171 shipyard workers. The effect of age on hearing was corrected with Robinson's model for an audiologically screened population. The exposure of the subjects to noise and their history of tobacco smoking were examined, with special reference to blood pressure and occupational Raynaud's phenomenon. Smoking without the presence of any other risk factors did not increase the risk for sensory neural hearing loss, but smoking in combination with elevated blood pressure and occupational Raynaud's phenomenon put workers at higher risk for hearing loss than any of these factors alone. PMID- 10380733 TI - Audioscan testing in patients with King-Kopetzky syndrome. AB - In this study, the Audioscan test has been used to detect early signs of hearing abnormalities in 80 patients with King-Kopetzky syndrome. A significantly higher prevalence of Audioscan notches between 500 and 3,000 Hz was found for each age band and gender in patients with King-Kopetzky syndrome than in control subjects. This indicates that Audioscan notches between 500 and 3,000 Hz may represent a fine hearing deficit as an indicator of mild auditory dysfunction in patients with King-Kopetzky syndrome. However, there was no significant difference in the percentage of notches in the 3,001-8,000 Hz frequency band between the King Kopetzky syndrome and control groups. PMID- 10380735 TI - Changes in serum osmolarity influence the function of outer hair cells. AB - Fast motility of outer hair cells (OHC) is thought to be based on a hydromechanic principle. In vitro, the function of OHCs can be disturbed by a change in the osmolarity of the culture medium. Whether changes in the serum osmolarity in vivo can also interfere with OHC motility has not been investigated as yet. Serum osmolarity of New Zealand White rabbits (n = 18) was elevated by a continuous infusion of glucose 40%, decreased by an infusion of aqua dest, or kept constant by an infusion of saline. OHC function was monitored using distortion products of otoacoustic emissions (DPOAE). Input output curves were established between 2 and 5 kHz (geometric mean of f2) with primaries of levels between 35 and 55 dB SPL. Cochlear perfusion was measured using a fluorescence microsphere method. Elevation of the serum osmolarity from 306 +/- 17 mosm/l to 365 +/- 23 induced a decrease of DPOAE between 3 and 12 dB SPL. Cochlear blood flow increased from 0.11 +/- 0.09 to 0.15 +/- 0.10 ml/min/g. When decreasing the serum osmolarity from 303 +/- 9 to 281 +/- 8 mosm/l, only slight changes of the DPOAE could be verified. As in the control group, cochlear perfusion was almost unchanged. In the control group, neither serum osmolarity nor DPOAE changed. Comparable to findings in vitro, increasing the serum osmolarity can lead to a disturbance of OHC function. In patients suffering from sudden hearing loss. dehydration due to physical or mental stress is often observed. This new and promising pathophysiological concept needs further clinical evaluation. PMID- 10380734 TI - Effect of white noise "masking" on vestibular evoked potentials recorded using different stimulus modalities. AB - Short latency vestibular evoked potentials (VsEPs) to linear acceleration impulses (L-VsEPs) are initiated in the otolith organs (saccule and utricle). Some of the saccule afferents have been reported to respond not only to linear acceleration, but also to high intensity acoustic stimuli. If so, the L-VsEP recorded from the saccule (elicited with the stimulus orientated relative to the head so as to optimally activate the saccule, i.e. stimulus in the vertical plane, Z-VsEP) should be reduced during high intensity broad band noise (BBN) "masking". Conversely, the utricular afferents have been reported to be less auditory-sensitive. Therefore, an L-VsEP which is mainly utricular in origin (stimulus in the horizontal plane, X-VsEP) should be less affected by this noise "masking". This was investigated in rats by recording X-VsEPs and Z-VsEPs and angular VsEPs (A-VsEPs), originating in the lateral semi-circular canals, before, during and after exposure to short duration, high intensity (113 dB SPL) BBN. This intensity completely masked auditory nerve evoked responses. The Z-VsEP did appear to be slightly more affected by the noise "masking" than the X-VsEP, implying the presence of more auditory-sensitive elements in the saccule. The A VsEP was also affected by the BBN. The overall effect was relatively small (on average, 10-25% depression of the first wave of the different VsEPs). The responses showed recovery 5 min later. PMID- 10380736 TI - Cell proliferation in spiral ligament of mouse cochlea damaged by dihydrostreptomycin sulfate. AB - Cell proliferation of the spiral ligament in the normal and drug-induced damaged mice cochleae was investigated using the mitotic tracer bromodcoxyuridine (BrdU). Only a few nuclei labelled by BrdU were seen in the spiral ligament of the control mouse. However, many BrdU labelled nuclei in the spiral ligament in the cochlea damaged by dihydrostreptomycin sulfate were found. The expression of fibroblast growth factor receptor and connexin 43 was detected in the spiral ligament where BrdU labelled cells were found. These results suggest that cell proliferation in the spiral ligament may occur after the drug-induced damage, and this process is probably related to the recovery of cochlear function. PMID- 10380737 TI - Spatial sound detection and the role of the inferior colliculus in the Long-Evans rat. AB - The ability of Long-Evans hooded rats (n = 10) to detect sounds presented from sources in the horizontal plane at 0 degrees elevation and the effects of bilateral lesions of the inferior colliculus on these abilities were examined. Rats were trained on a directional detection task which required animals to suppress licking responses in a conditioned avoidance paradigm when 100-ms noise bursts were presented at random from speakers at 45 degrees intervals beginning at azimuth (0 degrees). A task performance rate was determined by reducing the correct lick suppression rate for signal trials by the proportion of incorrect suppression responses on non-signal trials. Higher performance rates were observed for stimuli presented from 0-90 degrees than for stimuli presented in the caudal hemifield prior to surgical procedures. Bilateral lesions restricted to the inferior colliculus reduced detection performance (p < 0.05) and shifted the best performance rates from sounds presented at 0-45 degrees to stimuli emitted from a 90 degrees source (p < 0.05). These results demonstrate that pigmented rats show differential detection levels for noise bursts presented from different locations throughout the horizontal interaural plane, and suggest that the inferior colliculus is involved in this aspect of directional hearing. PMID- 10380738 TI - On inner ear function and the origin of oto-acoustic emissions. AB - The extremely low hearing threshold of the mammalian ear suggests the presence of a special amplifying mechanism, because the stereocilia of the outer hair cells (OHCs) are not likely to be sensitive enough themselves, although their mechanical embedding may provide some amplification. In the past decades, biophysicists have increasingly turned to the chaos theory for explanation. a theory the implications of which are considerable. One of its major tenets, self organization, is not easily understood at first glance, but is easily reproducible mathematically. With self-organization, the processes involving the OHCs can readily be simulated: Self-organization can help to explain why OHCs vibrate at amplitudes much higher than those of the exciting stimulus. To further our understanding of the process of hearing, vibratory processes, which presumably occur in normal and damaged OHC clusters, are described and compared with a mathematical analysis of data sets obtained from normal subjects using an extremely sensitive microphone. PMID- 10380739 TI - Ultrastructural localization of glucose transporter 1 (GLUT1) in guinea pig stria vascularis and vestibular dark cell areas: an immunogold study. AB - Glucose transporter 1 (GLUT1) is one of the facilitated-diffusion glucose transporters, which plays a major role in the glucose transport across blood tissue barriers. We demonstrated localization of GLUT1 in guinea pig stria vascularis (SV) and vestibular dark cell areas, using electron microscopy. GLUT1 immunoreactivity was seen mainly in the SV. Electron microscopically, immunogold particles were localized along the plasma membranes of the basal cells and the basolateral infoldings of the marginal cells. Erythrocyte membranes and the capillary walls in both the SV and the dark cell areas were also positive for GLUT1. These results indicate that GLUT1 is involved in glucose regulation for energy metabolism in the marginal cells and vestibular dark cells, and in glucose transport between blood and perilymph. PMID- 10380740 TI - The rebound phenomenon of glycerol-induced changes in the endolymphatic space. AB - Volumetric changes of the scala media were histologically investigated in normal guinea pigs to see whether glycerol-induced volumetric change of the scala media followed a biphasic course similar to the auditory threshold in the glycerol test. Glycerol was administered orally in a 12 ml/kg dose. The volume of the scala media was assessed by examining the cross-sectional area of the scala media in the mid-modiolar sections of the cochlea. Histological study revealed that the time-course of the change in the volume of the scala media after glycerol intake showed biphasic changes. Specifically, the early phase of the glycerol effect is a decrease in endolymph volume. The volume of the scala media significantly decreased by 11.4+/-2.9% 2 h after glycerol intake. Thereafter, the volume began to increase, and reached its peak 6-12 h after intake. In addition, the volume of the scala media significantly increased by 17.6+/-1.1% after 6 h. The present study indicated that the secondary increase in the volume of the scala media following glycerol intake played an important role in the rebound phenomenon in the glycerol test, although the mechanism underlying the hearing loss with the endolymphatic hydrops remains to be elucidated. PMID- 10380741 TI - Ultrastructure of the endolymphatic sac in two-phase endolymphatic hydrops in the guinea pig. AB - Two-phase endolymphatic hydrops is a subtle experimental model for Meniere's disease. Chronic dysfunction of the endolymphatic sac, induced by dissection of the most distal part without causing damage to the intermediate part, is combined with increased endolymph production induced by administration of aldosterone which stimulates the N/K-ATPase in the stria vascularis. A transmission electron microscopic study was performed on the endolymphatic sacs of four groups of guinea pig cochleas: controls: non-operated aldosterone-treated cochleas; operated (dissection of the endolymphatic sac) cochleas; operated and aldosterone treated cochleas. Light and electron microscopy showed a normal morphology in the controls. Aldosterone treatment had no visible effect. Dissected ears revealed severe deviations. The epithelium of the intermediate sac was low, showed dilated lateral intercellular spaces indicating elevated fluid transport and displayed serious degenerative processes. Distally, the endolymphatic sac was completely blocked by newly formed bone. Additional aldosterone treatment had no cumulative effect on the dissected ears. PMID- 10380742 TI - Effects of nasal obstruction on Eustachian tube function and middle ear pressure. AB - To evaluate the relationship between nasal obstruction and otitis media, 10 ferrets were studied before and after either unilateral (E = 5) or bilateral (n = 5) nasal obstruction. Observations included otomicroscopic assessments of middle ear status, tympanometric recordings of middle ear pressure and forced-response, inflation-deflation and continuous monitoring tests of Eustachian tube function. During the 6 8 week post-obstruction follow-up period no animal developed evidence of otitis media. Abnormal positive middle ear pressures lasting for the period of follow-up occurred only in the animals with bilateral nasal obstruction. Eustachian tube function test results showed these pressures to be generated during swallowing. No changes in the passive function of the tube were documented in either group, but changes in active function consistent with alterations in the pressure gradient between the middle ear and the nasopharynx were observed in both groups. PMID- 10380743 TI - Analysis of the best site on the stapes footplate for ossicular chain reconstruction. AB - Experiments were performed in 22 fresh human temporal bones to compare the relative acoustic function of three stapes footplate sites for an incus stapes superstructure replacement prosthesis (I-SRP). The three sites evaluated were the anterior, centre and posterior footplates. A new round window (RW) measurement method was used to make the comparisons. A small glass microsphere was placed in the centre of the RW as a target. A Polytec laser Doppler vibrometer was used to measure round window displacement in response to 50 pure tones between 200 and 10,000 Hz presented at 80 dB SPL at the tympanic membrane (TM). After a baseline measurement of RW displacement in the intact temporal bone, the incus was removed and a cement I-SRP (CIRP) formed between the mid-malleus handle and each of the three test footplate sites, in random order. RW displacement was again measured after placement of the CIRP at each of three sites. We found the centre site to be 3.0-7.0 dB better than the anterior site above 2,000 Hz. There were no differences between the anterior and centre sites below 2,000 Hz. The posterior site was the worst at all frequencies. PMID- 10380744 TI - Topical budesonide treatment reduces endothelial expression of intercellular adhesion molecules (vascular cell adhesion molecule-1 and P-selectin) and eosinophil infiltration in nasal polyps. AB - Infiltration of eosinophil granulocytes and endothelial expression of vascular cell adhesion molecule-1 (VCAM-1) and P-selectin was investigated in biopsies of polyps and inferior turbinates from 16 patients with nasal polyps, by the use of immunohistochemical staining and stereological quantification before, during and after topical treatment with budesonide (Rhinocort Turbuhaler). Before glucocorticoid treatment a higher density of eosinophil profiles (p < 0.005), making up a larger proportion of the total cellular infiltrate (p < 0.0005), was found in the polyps compared with the inferior turbinates. Endothelial VCAM-1 expression was higher in polyps than in inferior turbinates (p < 0.005), in contrast with the expression of P-selectin, which was more frequently expressed in the inferior turbinates (p < 0.05). Topical glucocorticoid treatment reduced the density of eosinophil profiles (p < 0.05) and the endothelial expression of VCAM-1 (p < 0.007) and P-selectin (p < 0.02) in polyps. Eosinophil counts and VCAM-1 expression returned to pre-treatment levels 8 weeks after discontinuation of budesonide treatment. The observed reduction in endothelial expression of cellular adhesion molecules may interfere with cellular recruitment in nasal polyps and thus contribute to the known anti-inflammatory effect of glucocorticoid in nasal polyposis. PMID- 10380745 TI - Effect of bupivacaine on pain after tonsillectomy: a randomized clinical trial. AB - Several authors have found that pre-incisional injection of local anaesthetics reduces postoperative pain. In the present double-blind study, comprising 126 inpatients aged 6-42 (mean 19) years, we investigated whether pre-incisional injection of bupivacaine during general anaesthesia reduces the pain experienced after tonsillectomy. The patients were randomized into three treatment groups: 43 patients were injected with 5 ml of bupivacaine (2.5 mg/ml)+ epinephrine (5 microg/ml) solution in both tonsillar fossa, 41 had epinephrine (5 microg/ml) + saline (9 mg/ml) and 42 patients received saline (9 mg/ml) only. Self-assessment of pain during the first postoperative week (repeated measures) was recorded. Use of analgetics, experience of the surgeons, peroperative bleeding and several other clinical parameters were assessed. Analyses of covariance with repeated measures was carried out for each pain score. In general there was no statistical significant difference in pain scores, represented by a visual analogue scale (VAS) between the three treatment groups. However, injection of bupivacaine into the tonsillar fossa seemed to reduce pain shortly after the operation in the age group 19-24 years. Further, females and older patients reported more pain and used more analgetics than males and younger patients. Increasing experience of the surgeon was related to a lower score for baseline pain shortly after the operation. Epinephrine in bupivacaine or saline reduced peroperative bleeding. We conclude that bupivacaine does not provide significant postoperative analgesia after tonsillectomy in an unselected group of patients. PMID- 10380746 TI - Cellular immune response of adenoidal and tonsillar lymphocytes to the P6 outer membrane protein of non-typeable Haemophilus influenzae and its relation to otitis media. AB - Cellular immune responses to the P6 outer membrane protein of non-typeable Haemophilus influenzae (NTHi) were determined in vitro by measuring immunoglobulin (Ig) secreting cells and lymphocyte proliferation in adenoidal and tonsillar lymphocytes from 19 children. Preliminary tests showed that P6 did not stimulate naive cells such as cord blood lymphocytes, but did stimulate sensitized cells in adenoids and tonsils. Cellular proliferation was significantly higher in adenoidal lymphocytes than in tonsillar lymphocytes (median: quadratile of stimulation index = 3.7:2.3-5.5 vs. 1.2:1.0-2.1, p < 0.02). A comparison between children with or without otitis media revealed that proliferative responses to P6 of adenoidal lymphocytes from children with otitis media were significantly decreased (2.0:1.8-3.6 vs. 3.7:2.3-5.5, p < 0.04). P6 specific antibody secreting cells were identified in a total of 14 adenoids and the number of cells secreting IgA was decreased in the otitis media group compared to controls (median: quadratile/10(6) cells = 435:359-499 vs. 755:593 1870, p < 0.05). Cultivation with P6 stimulated IgA secretion in children without otitis media, while no response was seen in children with otitis media (median: quadratile/10(6) cells = 1323:915-2410 vs. 2240:1900-2830, p < 0.02). These preliminary data demonstrate that lymphocytes from adenoids and tonsils recognize P6 as a specific antigen and that the adenoid is the more reactive of the two organs. Impaired P6-specific cellular immune responses of adenoids in children with otitis media may explain the recurrent nature of otitis media due to NTHi in the otitis prone population. PMID- 10380747 TI - Increased interleukin-6, interferon-gamma and tumour necrosis factor-alpha production by tonsillar mononuclear cells stimulated with alpha-streptococci in patients with pustulosis palmaris et plantaris. AB - In vitro cytokine production by tonsillar mononuclear cells under culture conditions with or without lyophilized streptococcal antigens was measured by enzyme-linked immunosorbent assay (ELISA). Under culture conditions without any stimulus, a certain amount of interferon (IFN)-gamma, tumour necrosis factor (TNF)-alpha, and interleukin (IL)-6, was detected in the supernatant culture fluids from all 29 donors tested and just detectable levels of IL-1alpha and IL-2 were identified in 5 of 12 donors tested; but IL-4 and IL-5 were not detected in any supernatants from 21 donors tested. In 16 patients with pustulosis palmaris et plantaris (PPP), all three alpha-streptococcal antigens from Streptococcus, (S.) sanguis, S. salivarius and S. mitis induced production of IL-6. IFN-gamma and TNF-alpha by tonsillar mononuclear cells at a significant level. However, no significant induction of such cytokines by alpha-streptococcal stimulus was seen in 13 donors without PPP. No significant difference in cytokine production by tonsillar mononuclear cells was seen between the PPP and non-PPP subjects, when stimulated with S. pyogenes, Staphylococcus aureus Cowan I and pokeweed mitogen. In the experiments using tonsillar cell subsets, the cytokine inductions by alpha streptococcal stimulus were found in monocyte-depleted cells as well as in purified T-cells. but not in B-cells. These data suggest that IL-6, IFN-gamma and TNF-alpha may play an important role in immunological reactions in the tonsils and that a hyper-immune response to some of alpha-streptococcal antigens through increased production of such cytokines by tonsillar mononuclear cells, to which T cells mainly contribute, may play a key role in the pathogenesis of PPP. PMID- 10380748 TI - C-erbB-2 immunostaining in laryngeal cancer. AB - Tumour progression is strongly associated with a series of specific genetic changes in protooncogenes and tumour suppressor genes. One of the potential factors involved in tumorogenesis of squamous cell carcinomas is protooncogene c erbB-2 (also known as neu or HER2). The authors analysed the expression of c-erbB 2 oncoprotein in 154 cases of laryngeal squamous cell carcinomas and its relationship to the clinical outcome of the patients. The difference in c-erbB-2 oncoprotein expression between the control group and cancer patients was on the statistical borderline (p = 0.0470). There was no significant correlation between c-erbB-2 expression and sex and age of the patients. T stage, lymph node status, site and histopathological grading of the tumour and clinical outcome of the patients. Univariate analysis revealed no correlation between c-erbB-2 expression and survival rates. We conclude that immunohistological examination of c-erbB-2 on paraffin section is not a valuable prognostic factor in laryngeal carcinoma. PMID- 10380749 TI - Expression of myosin heavy chain mRNA in rat laryngeal muscles. AB - The composition of myosin heavy chain mRNA was analysed quantitatively in 5 intrinsic laryngeal muscles of rats, using a competitive polymerase chain reaction. Intrinsic laryngeal muscles with the fastest contraction times, e.g. ventricular thyroarytenoid muscle. lateral cricoarytenoid muscle. and vocalis muscle, contained 2 fast isoforms, comprising mainly type 2B myosin heavy chains (52.1, 44.6 and 8.2%, respectively) and type 2X myosin heavy chains (21.9, 37.6 and 80.8%, respectively). Conversely, muscles with slower contraction times, such as posterior cricoarytenoid muscle and cricothyroid muscle, contained more than 85% of 2 fast isoforms; mainly type 2X myosin heavy chains (52.4-72.1%, respectively) and type 2A myosin heavy chains (34.6-25.2%, respectively). The results show a strong correlation between the composition of fast myosin heavy chain isoforms and muscle contraction times. Type 2L myosin heavy chain transcripts specific for laryngeal muscles and extra-ocular muscles were expressed in the order of ventricular thyroarytenoid (9.5%) > lateral cricoarytenoid (4.8%) > vocalis (2.5%) > posterior cricoarytenoid muscle (0.9%), but were not expressed in cricothyroid muscle. Neonatal myosin heavy chain was also expressed in all laryngeal muscles, ranging from 0.04 to 3%, but embryonic myosin heavy chain was expressed in ventricular thyroarytenoid, posterior cricoarytenoid and cricothyroid muscle at very low levels. These results suggest that intrinsic laryngeal muscles have different expression patterns for myosin heavy chain isoforms and may have different regulatory roles related to their functional requirement. PMID- 10380751 TI - Enzymatic and chemical cleavage methods. AB - Cleavage-based methods of mutation detection offer a simple and intuitive means to detect and in most cases locate mutations within DNA fragment sizes ranging from 500 to 1500 bases. Their main advantages as a presequencing screening technology when scanning for unknown mutations is the potential to increase throughput by multiplexing. Combined with lower reagent costs per sample, mutation scanning methods offer significant advantages over currently available sequencing techniques and are likely to be of increasing importance as genomic sequence data becomes more readily available. Although enzymatic methods offer the advantages of simpler and less hazardous protocols, at present the most robust cleavage methods are based around chemical methods. PMID- 10380752 TI - Comparison of detection platforms and post-polymerase chain reaction DNA purification methods for use in conjunction with Cleavase fragment length polymorphism analysis. AB - The removal of impurities and contaminants from PCR-amplified fragments is important for mutation detection methods which identify mutations based on shifts in electrophoretic mobility. This is particularly critical for assays and detection methods which use target DNA that is labeled prior to analysis and electrophoretic detection. We examined several procedures for purifying DNA amplified by the polymerase chain reaction (PCR) and their use in conjunction with a novel DNA scanning method, the Cleavase fragment length polymorphism (CFLP)* assay. In this study, a 480 bp DNA fragment, fluorescently labeled on the 5'-end of one strand, was amplified and subjected to various widely used purification procedures, including several commercially available clean-up kits. We demonstrate that visualization of the fluorescent label, as opposed to simple ethidium bromide staining, reveals the presence of considerable levels of labeled, truncated, amplification products. The various procedures were evaluated on the basis of their ability to remove these unwanted DNA fragments as well as on the degree to which they inhibited or promoted the CFLP reaction. Several procedures are recommended for use with CFLP analysis, including isopropanol precipitation, gel excision, and several commercially available spin columns. Concurrently, we evaluated (compared) a number of commonly used visualization platforms, including fluorescence imaging, chemiluminescence, and post electrophoretic staining, for the ability to detect CFLP pattern changes. The advantages and disadvantages of different methods are discussed and amounts of DNA to be used for CFLP analysis on different detection platforms are recommended. PMID- 10380753 TI - Mutation identification DNA analysis system (MIDAS) for detection of known mutations. AB - We introduce a novel experimental strategy for DNA mutation detection named the Mismatch Identification DNA Analysis System (MIDAS) [1, 2], which has an associated isothermal probe amplification step to increase target DNA detection sensitivity to attomole levels. MIDAS exploits DNA glycosylases to remove the sugar moiety on one strand (the probe strand) at a DNA base pair mismatch. The resulting apyrimidinic/ apurinic (AP) site is cleaved by AP endonucleases/lyases either associated with the DNA glycosylase or externally added to the reaction mixture. MIDAS utilizes 32p- or FITC-labeled oligonucleotides as mutation probes. Generally between 20-50 nucleotides in length, the probe hybridizes to the target sequence at the reaction temperature. Mismatch repair enzymes (MREs) then cut the probe at the point of mismatch. Once the probe is cleaved, the fragments become thermally unstable and fall off the target, thereby allowing another full-length probe to hybridize. This oscillating process amplifies the signal (cleaved probe). Cleavage products can be detected by electrophoretic separation followed by autoradiography, or by laser-induced fluorescence-capillary electrophoresis (LIF-CE) of fluorophore-labeled probes in two minutes using a novel CE matrix. In the present experiments, we employed the mesophilic Escherichia coli enzyme deoxyinosine 3'-endonuclease (Endo V), and a novel thermostable T/G DNA glycosylase, TDG mismatch repair enzyme (TDG-MRE). MIDAS differentiated between a clinical sample BRCA 1 wild-type sequence and a BRCA1 185delAG mutation without the need for polymerase chain reaction (PCR). The combination of MIDAS with LIF CE should make detection of known point mutations, deletions, and insertions a rapid and cost-effective technique well suited for automation. PMID- 10380754 TI - Cleavage of double-stranded copy RNA by RNase 1 and RNase T1 provides a robust means to detect p53 gene mutations in clinical specimens. AB - Detecting somatic mutations in patient specimens is challenging because of the wide variation in quality and quantity of genomic DNA in clinically derived material. In cancer specimens, the challenge of detecting mutations is usually compounded by the presence of large numbers of nonmutated normal cells that dampen the relative signal that can be obtained from employing any mutation detection strategy. In the case of somatic mutations in the gene encoding the tumor suppressor, p53, a clinically useful mutation detection assay must be able to detect a wide variety of types of mutations scattered over five coding exons and their flanking intron sequences. This study examined the ability of a mutation detection strategy, termed NIRCA, to identify single-base mutations in the clinically relevant domain of the p53 gene. This strategy relies on RNase digestion-mediated cleavage of double-stranded copy RNA transcribed in vitro from polymerase chain reaction (PCR)-amplified genomic templates to detect mismatched base pairs resulting from hybridization of complimenting mutant and wild-type copy RNA strands. This assay system was found to robustly detect all twelve possible mismatches and the plus one and minus one frame shifts. Furthermore, the assay could detect mutations in clinical specimens when the mutant alleles composed as few as 4% of the total population of alleles isolated in bulk specimen genomic DNA. This mutation detection strategy worked efficiently in bladder, breast, colon and lung tumors as well as sediments from bladder cytology specimens. PMID- 10380755 TI - Mutation detection using fluorescent enzyme mismatch cleavage with T4 endonuclease VII. AB - Mutation detection techniques are often limited by sensitivity, ease of use and short fragment lengths. Enzyme mismatch cleavage (EMC) is a technique capable of rapidly scanning 1 kbp fragments of DNA for mutations. It relies on the ability of a bacteriophage resolvase enzyme, T4 endonuclease VII, to cleave DNA at single base pair mismatches and small heteroduplex loops. Originally the process was performed using radioactively labeled DNA and the results analysed after denaturing polyacrylamide gel electrophoresis and autoradiography. However, access to systems capable of detecting fluorescent species migrating through a gel and the widespread availability of fluorescently tagged primers have greatly improved upon the original technique. A number of mutations were detected using fluorescent EMC and the results compared to performing the technique using radiolabeled DNA. Fluorescent EMC detected the presence, position and number of mutations in DNA fragments as large as 1 kbp. The fluorescent method was found to have advantages over the original method in its ease of use, increase in signal to-noise ratio and the ability to multiplex samples by labeling DNA fragments with different fluorophores. This improvement on an already established method provides a sensitive, robust technique for mutation detection. PMID- 10380756 TI - Rapid DNA mutation identification and fingerprinting using base excision sequence scanning. AB - Base excision sequence scanning (BESS) is a new polymerase chain reaction (PCR) based mutation scanning method that locates and identifies all DNA mutations. The BESS method consists of two procedures that generate "T" (BESS T-Scan) and "G" ladders (BESS G-Tracker) analogous to T and G ladders of dideoxy sequencing. The BESS procedures are simple to perform and require no special equipment or gels, no reaction optimization beyond PCR, and no heteroduplex formation. The samples are analyzed on standard sequencing gels or on automated DNA sequencers, and the data produced are easy to interpret, requiring a simple comparison of the sequence of normal and mutant DNA. The BESS method is versatile, having applications not only for mutation detection, but also single nucleotide polymorphism (SNP) discovery and analysis, DNA fingerprinting (including viral and bacterial typing), and clone identification. In this study, we utilize BESS in two of these applications: detection of a point mutation in BRCA1, and DNA typing of human papilloma virus (HPV). PMID- 10380757 TI - Single-strand conformation polymorphism and heteroduplex analysis for gel-based mutation detection. AB - Single-strand conformation polymorphism (SSCP) and heteroduplex analysis (HA) are popular electrophoretic methods for the identification of sequences. The principle reasons for the popularity of these two methods are their technical simplicity and their relatively high sensitivity for the detection of mutations. Here we review the theory and practice of SSCP and HA, including the factors contributing to the sensitivity of mutation detection. For SSCP analysis, these factors include: choice of gel matrix, electrophoretic conditions, presence of neutral additives, fragment size, and G+C content For HA, the principle factors influencing sensitivity are the gel matrix and the identity of the base mismatch. PMID- 10380758 TI - Use of a DNA toolbox for the characterization of mutation scanning methods. I: construction of the toolbox and evaluation of heteroduplex analysis. AB - A systematic characterization of the effects of important physical parameters on the sensitivity and specificity of methods in searching for unknown base changes (mutations or single nucleotide polymorphisms) over a relatively long DNA segment has not been previously reported. To this end, we have constructed a set of molecules of varying G+C content (40, 50, and 60% GC) having all possible base changes at a particular location - the "DNA toolbox". Exhaustive confirmatory sequencing demonstrated that there were no other base changes in any of the clones. Using this set of clones as polymerase chain reaction (PCR) templates, amplicons of various lengths with the same base mutated to all other bases were generated. The behavior of these constructs in manual and automated heteroduplex analysis was analyzed as a function of the size and overall base content of the fragment, the nature and location of the base change. Our results show that in heteroduplex analysis, the nature of the mismatched base pair is the overriding determinant for the ability to detect the mutation, regardless of fragment length, GC content, or the location of the mutation. PMID- 10380759 TI - Use of DNA toolbox for the characterization of mutation scanning methods. II: evaluation of single-strand conformation polymorphism analysis. AB - Single-strand conformation polymorphism (SSCP) is one of the most commonly used methods for searching for unknown base changes (mutations). In order to characterize systematically the effects of important physical parameters on the sensitivity and specificity of SSCP, we used the DNA toolbox constructed as described in the companion paper [2]. Using this set of DNA molecules as polymerase chain reaction (PCR) templates, amplicons of various lengths with the same base, mutated to all other bases, were generated. The behavior of these constructs in manual and automated SSCP was analyzed as a function of the size, overall base content of the fragment, nature and location of the base change, and the temperature and pH of electrophoresis. Our results demonstrate that all of these variables interact to determine the rate of detection of single-base changes, with the GC content being the predominant determinant of detection sensitivity. PMID- 10380760 TI - Detection of mutations and polymorphisms in the p53 tumor suppressor gene by single-strand conformation polymorphism analysis. AB - Deciphering the genetic mechanisms in cancer development requires analysis of a large number of tumors for consistent genetic alterations. Single-strand conformational polymorphism (SSCP) analysis is a fast and efficient method for detecting mutations, deletions, insertions and loss of alleles. The primary advantage of this method is speed and ability to screen a large number of samples at one time. Here we report the use of the SSCP technique for rapidly screening tumor and normal tissues for mutations and polymorphisms in the p53 tumor suppressor gene. Because the DNA extracted from specific aberrant bands from different samples always give rise to the same nucleotide sequence upon sequencing analysis, the SSCP technique can be used as a diagnostic tool to identify the presence of such genetic alterations without having to spend time on further sequencing analysis. PMID- 10380761 TI - Detection of p53 gene mutations: analysis by single-strand conformation polymorphism and Cleavase fragment length polymorphism. AB - We have generated a collection of clones containing single point mutations within the exon 5-9 hot spot regions of the p53 gene by using polymerase chain reaction (PCR) to amplify select regions of the gene from characterized cell lines. These clones were then used to address the sensitivity of mutation detection using slab gel single-strand conformation polymorphism (SSCP) and Cleavase fragment length polymorphism (CFLP) assay systems. Both methods exhibited high sensitivities for the detection of mutations in cloned p53 mutations in this study: 97% for CFLP and 94% for SSCP. In addition to resulting in higher sensitivity of mutation detection, CFLP has the capability to analyze longer fragments. In this study, CFLP identified five intronic mutations which were not investigated in the exon specific SSCP assay. These results agree with those found elsewhere and demonstrate that CFLP scanning can have practical advantages when used for the identification of sequence alterations within the p53 gene. PMID- 10380762 TI - Applications of constant denaturant capillary electrophoresis/high-fidelity polymerase chain reaction to human genetic analysis. AB - Constant denaturant capillary electrophoresis (CDCE) permits high-resolution separation of single-base variations occurring in an approximately 100 bp isomelting DNA sequence based on their differential melting temperatures. By coupling CDCE for highly efficient enrichment of mutants with high-fidelity polymerase chain reaction (hifi PCR), we have developed an analytical approach to detecting point mutations at frequencies equal to or greater than 10(-6) in human genomic DNA. In this article, we present several applications of this approach in human genetic studies. We have measured the point mutational spectra of a 100 bp mitochondrial DNA sequence in human tissues and cultured cells. The observations have led to the conclusion that the primary causes of mutation in human mitochondrial DNA are spontaneous in origin. In the course of studying the mitochondrial somatic mutations, we have also identified several nuclear pseudogenes homologous to the analyzed mitochondrial DNA fragment. Recently, through developments of the means to isolate the desired target sequences from bulk genomic DNA and to increase the loading capacity of CDCE, we have extended the CDCE/hifi PCR approach to study a chemically induced mutational spectrum in a single-copy nuclear sequence. Future applications of the CDCE/hifi PCR approach to human genetic analysis include studies of somatic mitochondrial mutations with respect to aging, measurement of mutational spectra of nuclear genes in healthy human tissues and population screening for disease-associated single nucleotide polymorphisms (SNPs) in large pooled samples. PMID- 10380763 TI - Horizontal two-dimensional electrophoresis of complex DNA samples using precast gel systems. AB - We have developed a simplified procedure for the separation of enzyme-digested genomic DNA, or of complex mixtures of cloned DNA, into two dimensions. The procedure relies on the use of precast gels for horizontal electrophoretic separations. Precast agarose-type gels are used for the first-dimensional separation of fragments based on size. Precast polyacrylamide gels are used for the second-dimensional separation of fragments. The separated fragments are subjected to enzymatic digestion in situ prior to their transfer to the second dimensional gel. Applications of this procedure include the analysis of DNA libraries, analysis of yeast artificial chromosomes (YACs) as well as other similar preparations, and the screening of genomic DNA for the occurrence of multi-copy DNA fragments as in the case of genomic amplifications in cancer. PMID- 10380764 TI - Two-dimensional gene scanning: exploring human genetic variability. AB - Current methods for mutation detection are not optimized for the generation of highly accurate data on multiple genes of hundreds of individuals in population based studies. Two-dimensional gene scanning (TDGS) is a high-resolution system for detecting mutational variants in multiple genes in parallel. TDGS is based on a combination of extensive multiplex polymerase chain reaction (PCR) and two dimensional (2-D) DNA electrophoresis. The latter involves a size separation step followed by denaturing gradient gel electrophoresis (DGGE). TDGS tests for a number of large human disease genes have been designed, using a computer program to optimally position PCR primers around the relevant target sequences (e.g., exons) and evaluated using panels of samples with previously detected mutations. The results indicate a high sensitivity and specificity, equal to nucleotide sequencing, which is generally considered as the gold standard. Here, we describe the different components of the TDGS process and its potential application as a high-throughput system for the systematic identification of human gene variants. PMID- 10380765 TI - Microplate array diagonal gel electrophoresis for mutation research in DNA banks. AB - Molecular genetic epidemiology, association and linkage studies in populations, human or other species, is now yielding powerful new insights into disease and susceptibility genes. Inter alia, the subject requires laboratory analytical methodologies focused on achieving high throughput. Here we review one suite of methodology suitable for such laboratories. Microplate array diagonal gel electrophoresis (MADGE) was invented for molecular genetic epidemiological studies. It combines direct compatibility with microplates, convenient polyacrylamide gel electrophoresis and economy of time and reagents, at minimal capital cost, and enables one user to run up to several thousand gel lanes per day for direct assay of single-base variations. Melt-MADGE combines temporal thermal ramp apparatus to achieve similar throughput for de novo mutation scanning. PMID- 10380766 TI - Single nucleotide polymorphism determination using primer extension and time-of flight mass spectrometry. AB - The high frequency of single nucleotide polymorphisms (SNPs) in the human genome makes them a valuable source of genetic markers for identity testing, genome mapping, and medical diagnostics. Conventional technologies for detecting SNPs are laborious and time-consuming, often prohibiting large-scale analysis. A rapid, accurate, and cost-effective method is needed to meet the demands of a high-throughput DNA assay. We demonstrate here that analysis of these genetic markers can now be performed routinely in a rapid, automated, and high-throughput fashion using time-of-flight mass spectrometry and a primer extension assay with a novel cleavable primer. SNP genotyping by mass spectrometry involves detection of single-base extension products of a primer immediately adjacent to the SNP site. Measurement of the mass difference between the SNP primer and the extension peak reveals which nucleotide is present at the polymorphic site. The primer is designed such that its extension products can be purified and chemically released from the primer in an automated format. The reduction in size of the products as a result of this chemical cleavage allows more accurate identification of the polymorphic base, especially in samples from a heterozygotic population. All six possible heterozygotes are resolved unambiguously, including an A/T heterozygote with extension products differing by only 9 Da. Multiplex SNP determination is demonstrated by simultaneously probing multiple SNP sites from a single polymerase chain reaction (PCR) product as well as from multiplexed PCR amplicons. Samples are processed in parallel on a robotic workstation, and analyzed serially in an automated mass spectrometer with analysis times of only a few seconds per sample, making it possible to process thousands of samples per day. PMID- 10380767 TI - Novel application of PhastSystem polyacrylamide gel electrophoresis using restriction fragment length polymorphism--internal transcribed spacer patterns of individuals for molecular identification of entomopathogenic nematodes. AB - differences! [editorial] [editorial]onomic way of identifying and assigning nematodes to taxons, which had already been determined either by comparative sequence analysis of nuclear rDNA internal transcribed spacer (ITS) region or by other methods of molecular or conventional taxonomy, is provided. Molecular identification of entomopathogenic nematodes (EPN) can be upgraded by basing it on PhastSystem polyacrylamide gel electrophoresis (PAGE) analysis of restriction fragment length polymorphism (RFLP) patterns of polymerase chain reaction (PCR) amplified DNA derived from single nematodes of Steinernema or Heterorhabditis spp. Although analysis from single worms has previously been made on agarose gel, the resolution on PhastSystem PAGE gel is much higher. The DNA sequences selected for analysis were those constituting the internal transcribed spacer region between the 18S and 26S rDNA genes within the rRNA operon. RFLP analysis was carried out by gel electrophoresis on the PhastSystem (Pharmacia) as detailed elsewhere (Triga et al., Electrophoresis 1999, 20, 1272-1277. The downscaling from conventional agarose to PhastSystem gels resulted in pattern of DNA fragments differing from those obtained with agarose gel electrophoresis under conventional conditions by increasing the number of detected fragments. The approach supported previous species identifications and was able to identify several unclassified isolates, such as those from Hungary and Ireland, and provides a method for identification of previously unclassified strains. We confirmed that Heterorhabditis "Irish Type", represented by two strains of different geographical origin, comprise a species different from H. megidis. We also confirmed that strain IS5 belongs to the species H. indicus rather than to H. bacteriophora, as had been suggested previously. PMID- 10380768 TI - Gel electrophoretic restriction fragment length polymorphism analysis of DNA derived from individual nematodes, using the PhastSystem. AB - The DNA sequences constituting the internal transcribed spacer region, located between 18S and 26S rDNA genes within the rRNA operon, derived from single nematodes of two genera (Steinernema and Heterorhabditis) were amplified by polymerase chain reaction (PCR) and subjected to digestion by four restriction enzymes. The digests were analyzed by restriction fragment length polymorphism (RFLP) gel electrophoresis on the PhastSystem, using 7.5%T, 5%C(Bis) polyacrylamide. The downscaling from conventional agarose to PhastSystem gels permitted the analysis to be done on individual nematodes, rather than on mixed samples with average properties. The analysis time was reduced so as to allow for the electrophoretic separation on 200 samples/workday. The resulting patterns of DNA fragments differed from those obtained by agarose gel electrophoresis under conventional conditions by an increased number of detected fragments. The PhastSystem gel analysis provides the basis for taxonomical revisions. PMID- 10380769 TI - High performance DNA sequencing, and the detection of mutations and polymorphisms, on the Clipper sequencer. AB - The Visible Genetics Clipper sequencer is a new platform for automated DNA sequencing which employs disposable MicroCel cassettes and 50 microm thick polyacrylamide gels. Two DNA ladders can be analyzed simultaneously in each of 16 lanes on a gel, after labeling with far-red absorbing dyes such as Cy5 and Cy5.5. This allows a simultaneous bidirectional sequencing of four templates. We have evaluated the Clipper sequencer, by cycle-sequencing of an M13 single-stranded DNA standard, and by coupled amplification and sequencing (CLIP) of reverse transcribed human immunodeficiency virus (HIV-1) RNA standards and clinical patient samples. (i) Limitations of instrument. We have examined basic instrument parameters such as detector stability, background, digital sampling rate, and gain. With proper usage, the optical and electronic subsystems of the Clipper sequencer do not limit the data collection or sequence-determination processes. (ii) Limitations of gel performance. We have also examined the physics of DNA band separation on 50 microm thick MicroCel gels. We routinely obtain well resolved sequence which can be base-called with 98.5% accuracy to position approximately 450 on an 11 cm gel, and to position approximately 900 on a 25 cm gel. Resolution on 5 and 11 cm gels ultimately is limited by a sharp decrease in spacing between adjacent bands, in the biased reptation separation regime. Fick's (thermal) diffusion appears to be of minor importance on 6 cm or 11 cm gels, but becomes an additional resolution-limiting factor on 25 cm gels. (iii) Limitations of enzymology. Template quality, primer nesting, choice of DNA polymerase, and choice between dye primers and dye terminators are key determinants of the ability to detect mutations and polymorphisms on the Clipper sequencer, as on other DNA sequencers. When CLIP is used with dye-labeled primers and a DNA polymerase of the F667Y, delta(5'--> 3' exo) class, we can routinely detect single-nucleotide mutations and polymorphisms over the 0.35-0.65 heterozygosity range. We present an example of detecting therapeutically relevant mutations in a clinical HIV-1 RNA isolate. PMID- 10380770 TI - Inheritable defects in interleukin-12- and interferon-gamma-mediated immunity and the TH1/TH2 paradigm in man. PMID- 10380771 TI - Pityrosporum species as a cause of allergy and infection. PMID- 10380772 TI - Immune responses to Aspergillus antigen in IL-4-/-mice and the effect of eosinophil ablation. AB - BACKGROUND: Exposure to Aspergillus fumigatus allergens results in enhanced total serum IgE and peripheral blood eosinophils in mice. The associated pulmonary inflammation and immunologic responses are comparable to those detected in human allergic bronchopulmonary aspergillosis. Allergen-induced cytokines are thought to regulate the inflammatory and immune responses in these animals. METHODS: In the present study, we exposed C57BL/6 and BALB/c mice to A. fumigatus antigen. Both wild-type and IL-4 knockout phenotypes of animals of both strains were used. Some animals were also treated with anti-IL-5 or anti-IFN-gamma. Total serum IgE, Aspergillus species IgG subclass, peripheral blood eosinophils, and lung histology were studied. RESULTS: The results demonstrate similar lung inflammation in all wild-type and IL-4-/- animals exposed to A. fumigatus antigen. Similarly, in spite of the diverse immune response produced by the anticytokine treatment, no major differences were detected among any of the animal groups studied. CONCLUSIONS: It can be concluded that A. fumigatus exposure in an immunologically unaltered host is predominantly of a Th2 type, and that depletion of the Th2 cytokine leads to a similar lung inflammation but with a characteristic Th1 response, suggesting that the pathogenesis of allergic aspergillosis is the result of multiple induction pathways. PMID- 10380773 TI - Occurrence of allergic conditions in asthmatics with analgesic intolerance. AB - BACKGROUND: The study aimed to determine whether allergic conditions accompany analgesic intolerance. METHODS: A total of 132 analgesic-intolerant patients with bronchial asthma admitted to the adult allergy unit from January 1991 to October 1997 and 103 patients with bronchial asthma randomly selected from among the asthmatics referred to our department between January and October 1997 were enrolled in the study. Those having analgesic intolerance and bronchial asthma were accepted as group I; patients having only asthma were accepted as group II. A standard questionnaire was completed for all the patients. Physical examination, routine skin prick tests, determination of total IgE levels and blood type, and oral analgesic provocation tests were also performed. RESULTS: The results showed that some allergic conditions were significantly more common in group I (22.7% and 7.8% for food allergy/intolerance [P<0.05], 16.7% and 7.8% for antibiotic allergy, 16.7% and 2.9% for dermographism, 9.8% and 1.0% for metal allergy, and 9.1% and 1.0% for chronic urticaria for groups I and II, respectively [P<0.001]). In addition, the mean of the total IgE level in the serum was higher in group I than group II (77.6 and 53.7 IU/ml; P<0.05), and the cumulative analgesic consumption was more in group I (14.2+/-17.1 and 9.1+/-12.5 boxes; P<0.05). CONCLUSIONS: Dermographism; chronic urticaria; antibiotic, metal, and food allergy; high levels of total IgE; and a high amount of cumulative analgesic consumption may be the conditions accompanying analgesic intolerance in asthmatics. PMID- 10380774 TI - Basophil histamine release, IgE, eosinophil counts, ECP, and EPX are related to the severity of symptoms in seasonal allergic rhinitis. AB - BACKGROUND: Serum specific IgE, basophil histamine release, and blood eosinophil parameters are associated with allergic rhinitis, but investigations of the relationship to the severity of allergic symptoms are few and conflicting. Our study aimed to investigate the seasonal changes in the following laboratory tests: specific IgE, basophil histamine release, eosinophil counts, and serum and plasma eosinophil cationic protein (ECP) and eosinophil protein X (EPX), and to analyze, in detail, the relationship of each individual test to the severity of symptoms in rhinitis patients allergic to both birch and grass pollen. METHODS: The above tests were performed on blood samples obtained from 49 allergic rhinitis patients during the birch-pollen season, during the grass-pollen season, and after the seasons. Symptom-medication diaries were filled in during both pollen seasons. We used partial least square (PLS) analysis to establish an optimal statistical link between the symptom score and medication and the laboratory tests, in an investigator-independent way. RESULTS: Increases in specific IgE, basophil histamine release, eosinophil counts, serum ECP and EPX, and plasma EPX were observed from the birch-pollen season to the grass-pollen season, followed by a decrease from the grass-pollen season to after the pollen seasons, except for the specific IgE. No seasonal changes in plasma ECP and total IgE were seen. The PLS analysis found a relationship between symptom score and medication and the aggregate laboratory tests (F-test value 40.2, correlation 0.34 for the cumulative relation). However, the variation in laboratory tests could explain only half of the total variation in symptoms and less than a quarter of the total variation in medication. The symptom score and, to a minor degree, medication were especially correlated with the basophil histamine-release results, with a decreasing relevance of specific IgE, eosinophil counts, total IgE, serum and plasma EPX, and serum ECP. Plasma ECP was not related to the symptom score and medication. CONCLUSIONS: A significant relationship between the severity of allergic rhinitis and various allergic inflammatory markers was found but could account for only a minor part of the variation in the patients' evaluation of their disease. PMID- 10380775 TI - Determination of the allergenic activity of birch pollen and apple prick test solutions by measurement of beta-hexosaminidase release from RBL-2H3 cells. Comparison with classical methods in allergen standardization. AB - BACKGROUND: A murine in vitro model of the allergic type I reaction was set up to determine the biologic activity of extracts without involvement of human beings. It is based on beta-hexosaminidase release from passively sensitized RBL cells after allergen challenge. The intended application of this RBL cell assay in the field of quality control of allergenic extracts requires its comparison with established methods. METHODS: The activity of five standardized birch-pollen prick test solutions was determined in parallel by RBL assay, direct IgE binding, IgE-binding inhibition, major allergen content, histamine-release assay, and skin testing. RESULTS: The RBL cell-release assay corresponded well to other methods if a reagin raised against natural birch-pollen extract was used for passive sensitization. However, in the case of a reagin against recombinant Bet v 1, only a decreased activity was observed, presumably because a reduced number of epitopes were recognized by the monospecific reagin. In contrast to standardized birch-pollen extracts, nonstandardized apple extracts showed poor activity in all assays. CONCLUSIONS: This murine model might be a useful tool in the quality control of allergenic extracts. It combines properties of assays based on standardized antisera and of assays that consider IgE cross-linking properties. PMID- 10380776 TI - Allergen-induced accumulation of eosinophils and lymphocytes in skin chambers is associated with increased levels of interleukin-4 and sVCAM-1. AB - BACKGROUND: The aim of the study was to characterize the kinetic accumulation of various inflammatory mediators in allergen-challenged skin chambers applied on patients with pollen-related allergic rhinitis/mild asthma. METHODS: Skin blisters were induced on the forearms and challenged with allergen or phosphate buffered saline (PBS). Peripheral blood was drawn before and 8 h after challenge for analysis of differential cell counts, sVCAM-1, and alpha2-macroglobulin. Chamber fluids, collected at 1, 4, and 8 h after allergen application, were analyzed for differential cell counts, histamine, interleukin (IL)-4, sVCAM-1, and alpha2-macroglobulin. RESULTS: The number of recruited leukocytes was equal in allergen and PBS chambers; however, the numbers of eosinophils and lymphocytes were significantly (P< or =0.05) elevated in allergen-challenged chambers at 8 h. Compared to PBS chambers, allergen chambers contained significantly (P<0.01-0.05) higher levels of histamine (at 1 and 4 h), IL-4 (at 4 and 8 h), alpha2 macroglobulin (at 1 and 8 h), and sVCAM-1 (at 1 and 8 h). In contrast to alpha2 macroglobulin, levels of sVCAM-1 in peripheral blood were significantly (P<0.05) increased at 8 h. CONCLUSIONS: Increased levels of sVCAM-1 and IL-4 in allergen challenged chambers, in parallel with increased recruitment of eosinophils and lymphocytes, points to the participation of IL-4 and VCAM-1 in the development of the late-phase reaction. Increased levels of sVCAM-1 in allergen-challenged chambers probably reflects a combination of leakage and local production. PMID- 10380777 TI - Immunochemical detection of egg-white antigens and allergens in meat products. AB - BACKGROUND: The purpose of this study was to detect antigens and allergens in egg white byproduct ingredients and after their incorporation in processed pork meat pastes. Commercially prepared foods may have potentially allergenic ingredients (egg, milk, soybean, wheat, and peanut) added in processing. Since allergic patients may react to unidentified ingredients, it is important to assess the allergenic potency of these food proteins added during processing. Egg white was chosen as an experimental model, since egg is one of the most prevalent allergens in food hypersensitivity. METHODS: Experimental pork meat pastes containing egg white underwent pasteurization and sterilization. Ingredients derived from egg white or paste extracts were isoelectrofocused and then blotted onto cyanogen bromide-activated nitrocellulose membranes. Egg-white antigens were identified in ingredients and in meat products with rabbit anti-egg-white antiserum by isoelectric focusing immunoblotting. Allergens were identified with sera from sensitized patients. A sensitive ELISA test was developed to detect egg-white proteins in raw, pasteurized, and sterilized meat products. RESULTS: Antigens and allergens in four egg-white byproducts were detected. Egg-white antigens were detectable in all ingredients and meat pastes by ELISA. Allergens were detected in ingredients and in raw and pasteurized products by immunoprint techniques and ELISA. CONCLUSIONS: Masked egg-white allergens are recognized by human serum IgE after pasteurization. Egg-white antigens are detectable in sterilized meat by ELISA techniques. Ingestion of processed foods could entail a risk of allergic reactions for sensitized consumers. PMID- 10380778 TI - Association of heparin-induced skin lesions, intracutaneous tests, and heparin induced IgG. AB - BACKGROUND: Cutaneous heparin-induced allergic reactions to subcutaneous heparin may begin 2-5 days after administration. The relation of the delayed-type hypersensitivity and a systemic immunologic response is controversial. The present investigation aimed to analyze the occurrence of thromboembolic complication, pathologic heparin-induced platelet activation (HIPA), and the presence of circulating heparin-induced IgG in patients with heparin-induced skin reactions. METHODS: Intracutaneous tests, HIPA assay, and heparin-heparin IgG antibodies were performed in nine patients with heparin-induced skin lesions. RESULTS: Six of eight patients showed positive intracutaneous tests to heparin and to four low-molecular-weight heparins. Three of six heparin-positive patients presented hypersensitivity to a heparinoid, too. Two of three patients had a positive HIPA test and elevated heparin-induced IgG antibodies. Both patients developed complications presenting as heparin-induced skin necrosis or arterial thrombosis. Two of nine patients were treated with danaparoid, 4/9 patients received r-hirudin, and 1/9 received oral coumarin. In 2/9 patients, anticoagulant therapy was stopped, but these patients will receive r-hirudin if indicated. CONCLUSIONS: On the basis of the coincidence of local and systemic hyperreactivity to heparin and danaparoid, patients with heparin-induced skin lesions should receive r-hirudin, a nonheparin compound, for anticoagulant treatment. PMID- 10380780 TI - The effect of air filtration on airborne dog allergen. AB - BACKGROUND: Effective methods of reducing dog allergen are required to help alleviate symptoms in asthmatic patients sensitized to dog who refuse to part with their pet. The aim of this study was to investigate the use of the high efficiency particulate air (HEPA) filter air cleaner to reduce airborne Can f 1 in homes with a dog. METHODS: The effect of a HEPA air cleaner was investigated in nine homes with a dog. Samples were collected from two rooms of each house concurrently, one of which contained the dog, on two separate days (active day - HEPA air cleaner on - and control day). Eight consecutive 1-h samples were collected from each room with a high-volume air sampler (airflow rate 60 l/ min). Can f 1 was determined by monoclonal-polyclonal antibody-based ELISA. RESULTS: Baseline airborne Can f 1 levels were 3.8-fold greater when sampling was performed with a dog in the room (GM 27.1 ng Can f 1/m3, range 2.63-329) than when the dog was elsewhere in the house (GM 7.1 ng Can f 1/m3, range 0.69-27.2). When the dog was elsewhere in the house, airborne Can f 1 levels fell on both active and control days, but the magnitude of the reduction was significantly greater on the active days (P<0.05), and was approximately 90% from baseline. With the dog in the room, a significant fall in airborne Can f 1 was observed only on active days (75% from baseline), but not on control days (active vs control P<0.001). CONCLUSIONS: HEPA air cleaners reduce airborne Can f 1 in homes with dogs. Furthermore, preventing the access of the dog to the bedroom and possibly the living room may reduce the total allergen load inhaled. PMID- 10380779 TI - Immunologic significance of respirable atmospheric starch granules containing major birch allergen Bet v 1. AB - BACKGROUND: Birch-pollen allergens are an important cause of early spring hay fever and allergic asthma. Recently, we reported a mechanism for the release of respirable allergenic particles from birch pollen containing the major allergen Bet v 1. In this study, we aimed to assess the immunologic significance of the released Bet v 1-containing starch granules in the environment. METHODS: A two site monoclonal antibody-based assay (ELISA) was employed to quantitate Bet v 1 in high-volume air sampler filter extracts, and immunogold-labelling was used on sections of these extracts to localize Bet v 1. Immunoblot analyses were performed with pooled sera from patients sensitive to birch pollen. RESULTS: Atmospheric starch granules contained Bet v 1, and the concentration increased upon light rainfall. Sera from patients allergic to birch allergens recognized extracts from isolated starch granules. CONCLUSIONS: The clinical implications of these findings are that starch granules released from birch pollen are potentially able to trigger allergic asthmatic reactions to Bet v 1, since the allergen occurs in respirable particles. Thus, clinicians can advise asthma patients to remain indoors on days of light rainfall during the birch-pollen season to avoid high levels of allergen exposure. PMID- 10380781 TI - Leukotriene (LT)-receptor antagonist is more effective in asthmatic patients with a low baseline ratio of urinary LTE4 to 2,3-dinor-6-keto-prostaglandin (PG)F1alpha. AB - BACKGROUND: To test the hypothesis that urinary levels of arachidonic acid metabolites may be a predicting factor of the effects of pranlukast, a selective leukotriene (LT) antagonist, on chronic adult asthma, we investigated the relationship between its clinical efficacy and urinary eicosanoid levels. METHODS: An open, multicenter trial was conducted involving 38 stable moderate and severe asthmatic patients (mean percent predicted FEV1 was 71%). All patients received pranlukast (225 mg twice daily) for 4 weeks after a 2-week run-in period. Urinary levels of LTE4, 11-dehydro-thromboxane (TX) B2, 2,3-dinor-6-keto prostaglandin (PG) F1alpha, and creatinine were measured in 3-h urine collected on day 1 of the treatment. The responder was defined by an improvement of asthma symptom scores and peak expiratory flow rate (PEFR). RESULTS: One patient was excluded because of an adverse effect, nausea. Thirteen out of 37 subjects were responders and 24 were nonresponders. There were no significant differences in patients' backgrounds and urinary arachidonate levels between the two groups. The urinary LTE4 to 2,3-dinor-6-keto-PGF1alpha ratio in the responder was significantly lower (P=0.01) than that in the nonresponder. In all patients, a significant inverse correlation was revealed between the baseline urinary LTE4/2,3-dinor-6-keto-PGF1alpha ratio and the improvement of PEFR in the morning (r=-0.43, P=0.007). CONCLUSIONS: These data suggested that the urinary ratio of LTE4 to 2,3-dinor-6-keto-PGF1alpha might be one of the predictive markers of the clinical efficacy of this LT-receptor antagonist in asthmatic subjects. PMID- 10380782 TI - Provocations with perfume in the eyes induce airway symptoms in patients with sensory hyperreactivity. AB - BACKGROUND: In earlier studies, we have shown that patients with a history of sensory hyperreactivity develop asthma-like symptoms when exposed to strong scents, even if they cannot smell any scent. METHODS: For study of possible pathophysiologic mechanisms behind sensory hyperreactivity, the patients' airways and eyes were separately exposed to a common inducing factor, perfume. Eleven patients with a history of hyperreactivity to chemical trigger factors, such as perfume, were provoked single-blindly in a placebo-controlled, randomized study. During airway exposure, the eyes were covered and, during the eye exposure, the patients inhaled fresh air. A special face mask or a nose clip was used to avoid any smell. RESULTS: During the 30-min exposure to perfume, there was a gradual increase in three main symptoms; i.e., eye irritation, cough, and dyspnea, after both the airway and eye exposures. The increases were significant compared with placebo. CONCLUSIONS: Asthma-like and other symptoms, such as irritation of the eyes, may be induced by exposure of both the airways and the eyes in patients with sensory hyperreactivity. This points to the importance of studying the sensory nervous system, not only in the airways, but also in other organs. PMID- 10380783 TI - Predictive capacity of histamine release for the diagnosis of drug allergy. AB - BACKGROUND: The diagnosis of immediate allergic reactions to drugs is difficult, requiring in vitro test development. Basophils are likely to be involved in these reactions, and to evaluate the sensitivity, the specificity, and the predictive values of the histamine-release test, we performed a prospective study in 68 patients tested for suspected drug allergy. METHODS: Positive diagnosis was established by history, skin tests, and, if needed, oral provocation tests. Histamine release in the presence of the drug was assessed on heparinized whole blood by enzyme immunoassay (Immunotech, France), and the cutoff value was set at 5% of total histamine content. Spontaneous and anti-IgE-induced histamine release was also studied in all subjects. RESULTS: All patients presented to our clinic with reactions ranging from maculopapular exanthema to anaphylactic shock. Thirty five patients had proven drug allergy; 33 were not allergic to drugs and served as a control group together with 40 other subjects with no history of drug allergy. Net histamine release was positive in 18/35 allergics and 27/73 nonallergics, giving poor sensitivity (51.4%), specificity (63.0%), and positive predictive value (29.3%), but valuable negative predictive value (81.1%). CONCLUSIONS: The usefulness of the in vitro histamine-release test for the diagnosis of drug allergy appears to be insufficient. PMID- 10380784 TI - Cyclosporin A (CyA) reduces sCD30 serum levels in atopic dermatitis: a possible new immune intervention. AB - Atopic dermatitis (AD) is a chronic inflammatory skin disease frequently associated with asthma, rhinitis, and food allergy. Lymphocytes producing Th2 type cytokines (such as interleukin [IL]-3, IL-4, and IL-5) have been thought to have a key role in the pathogenesis of the disease. We have recently demonstrated that elevated serum levels of the soluble form of CD30 (sCD30), an activation marker of Th2-cell clones, correlates with disease activity in pediatric patients suffering from AD. Clinical trials have demonstrated that cyclosporin A (CyA) treatment resulted in significant improvement of clinical symptoms in patients affected with AD. In this study, we evaluated the role of CyA in modulating sCD30 release in a group of adult patients affected by severe AD treated with CyA at the dosage of 3.5 mg/kg body weight for 12 weeks. Our results demonstrated, in parallel with an improvement of clinical symptoms, a significant reduction of serum levels of both IL-4 and sCD30, thus suggesting that CyA can prevent the activation of Th2 cells observed in AD. PMID- 10380785 TI - Immediate allergic and nonallergic reactions to Christmas and Easter cacti. AB - BACKGROUND: Occupational exposure to Christmas cacti has been reported as a cause of type I allergy. Therefore, the prevalence of immediate-type mucosal and skin reactions related to cactus exposure was studied in 103 employees in a cactus nursery. METHODS: The study was based on a questionnaire followed by clinical examination, skin prick tests (SPT) with standard inhalant allergens and cacti, and a histamine-release test (HRT/Refix) using fresh cactus extracts as elicitor. RESULTS: The questionnaire was answered by 84 (82%) of the nursery employees, and 63 (61%) were interviewed and skin prick tested; 58 of these were tested with HRT/Refix. Furthermore, 22 healthy controls were included and tested in vivo and in vitro. Cactus-related contact urticaria and/or rhinoconjunctivitis were reported by 37% of the cactus workers. Based on a combination of positive history, positive SPT, and positive HRT/ Refix to cactus, 8% of the cactus workers were allergic to cacti. No noncactus workers or controls were allergic to cacti by these criteria. Testing with fresh cactus material elicited positive SPT and negative HRT/Refix in 27 nursery workers and controls, of whom 12 had immediate-type skin and mucosal symptoms. CONCLUSIONS: Christmas and Easter cacti seemed to be able to induce contact urticaria and rhinoconjunctivitis on both an immunologic and a nonimmunologic basis. Personal atopy was associated with positive reactions to cacti. PMID- 10380786 TI - Dietary assessment in five cases of allergic reactions due to gastroallergic anisakiasis. AB - BACKGROUND: Anisakis simplex can cause allergic reactions in sensitized patients. Some of these reactions are related to acute parasitism, as is shown in gastroallergic anisakiasis (anisakiasis with digestive and predominantly allergic symptoms). At present, a nonseafood diet is recommended for all patients with any kind of A. simplex allergy. We wished to confirm the clinical suspicion that patients with allergic symptoms after ingestion of raw or undercooked seafood who are sensitized to A. simplex, and diagnosed with gastroallergic anisakiasis, can tolerate the ingestion of seafood when the parasites are dead and noninfective. METHODS: We included patients diagnosed with gastroallergic anisakiasis (positive skin prick test or/and serum specific IgE to A. simplex, with one or more parasites found by gastroscopy in the stomach). Patients included in the study gave written, informed consent. Specimens of A. simplex about 2 cm long were selected, placed in capsules, and frozen at -20 degrees C for more than 48 h to make them noninfective. We administered 11 specimens to every patient at the hospital. If they tolerated the larvae, they were told to eat well-frozen seafood (-20 degrees C at least 48 h). After 6 months, the patients were re-evaluated. RESULTS: Five patients accepted the challenge with noninfective A. simplex larvae. All tolerated the noninfective larvae. After eating deep-frozen seafood for 6 months, no patient suffered a reaction. CONCLUSIONS: In gastroallergic anisakiasis, the antigens of the live parasite probably cause the allergic symptoms. Patients with this disease can tolerate deep-frozen seafood, in which the parasites are dead. PMID- 10380787 TI - Occupational asthma caused by champignon flies. AB - BACKGROUND: Occupational bronchial asthma in mushroom (champignon) workers is unusual, although reports on it appeared in 1938 and 1951; we have not found any others since those dates. Here we report the case of a 52-year-old man who works as a champignon cultivator. He suffered rhinoconjunctivitis and asthma attacks whenever he entered the champignon culture caves. We studied flies as a possible antigen source. We collected these insects from the growing sites in order to identify them, and then prepare an extract; the samples turned out to be of two families of insects of the order Diptera, 98% from the Phoridae family (Brachycera suborder) and 2% from the Sciaridae (Nematocera suborder). METHODS: Skin prick tests, conjunctival provocation tests, serum specific IgE, specific IgE-binding fractions in immunoblotting, and monitoring of PEFR (at work and off work) were performed. RESULTS: IgE-mediated hypersensitivity to these flies was demonstrated by skin prick test, conjunctival provocation test, serum specific IgE, and IgE-binding fractions in immunoblotting. Monitoring of PEFR both at work and off work showed a clear relationship between symptoms, or fall in PEFR, and the workplace. CONCLUSIONS: We report the case of a patient suffering from asthma and rhinoconjunctivitis caused by hypersensitivity to fly proteins. PMID- 10380788 TI - A case of sesame seed-induced anaphylaxis. PMID- 10380789 TI - Anaphylactic reactions to topical rifamycin. PMID- 10380790 TI - Histamine content of peanuts. PMID- 10380791 TI - Failure of montelukast to prevent anaphylaxis to diclofenac. PMID- 10380792 TI - Allergy to human seminal fluid: a case of self-diagnosis. PMID- 10380793 TI - Allergy to fresh dill. PMID- 10380794 TI - Allergic alveolitis from pine sawdust. PMID- 10380795 TI - Zinc-finger proteins: the classical zinc finger emerges in contemporary plant science. AB - TFIIIA-type zinc fingers have been found in a number of eucaryotic transcription factors as DNA-binding motifs. In plants, as many as 30 proteins have been reported that have either one, two, three or four zinc fingers. Plant zinc-finger proteins are characterized by long spacers of diverse lengths between adjacent fingers and a highly conserved sequence, QALGGH, located within a putative DNA contacting surface of each finger. In vitro DNA-binding experiments with two fingered proteins of petunia have revealed that these proteins bind to target DNA sequences in a manner that is distinctive from that of their animal counterparts: (1) they specifically recognize the spacing between two core sites in target DNA, (2) they have a unique base-determinant position. Regulatory functions have been assigned to some of the TFIIIA-type zinc finger proteins in Arabidopsis, petunia and chinese cabbage. SUPERMAN, AtZFP1, PetSPL3 and BcZFP1 have been implicated in the developmental regulation of various floral and vegetative organs, presumably through the control of cell division and/or expansion in particular cell types. Several anther-specific zinc-finger proteins in petunia are presumed to be involved in the regulation of gametogenesis in both reproductive and non reproductive tissues of anther. STZ and ZPT2-2 are implicated in the response of plants to or tolerance for various stresses. PMID- 10380796 TI - Novel characteristics and regulation of a divergent cinnamate 4-hydroxylase (CYP73A15) from French bean: engineering expression in yeast. AB - cDNAs showing high sequence similarity (>70%) over large stretches to plant CYP73A orthologues from other species were isolated from a cDNA library derived from mRNAs expressed in elicitor-treated suspension-cultured cells. These clones appear to code for a full-length 1554 bp open reading frame with a 78 bp 5' untranslated region and a 140 bp 3'-untranslated region. The open reading frame, determined by sequence similarity, codes for a protein with a predicted Mr of 59229 and a pI of 8.8. It contains the conserved cysteine haem-binding site found in all cytochrome P450s. The protein encoded by this cDNA diverges however from other CYP73As in its N- and C-terminus and in four domains internally, so that overall sequence similarity is in the range 58-66%. Many clones contained an identical intron, which may be associated with a novel regulatory mechanism. Sequence similarity is sufficient for it to be classified as CYP73A15, although it is the least similar member of this family classified so far. The cDNA was expressed in yeast. Successful expression of cinnamate 4-hydroxylase activity required removal of the intron. High-level expression also required modification of the N-terminus to that of CYP73A1. Yeast did not process the intron at all and the leader sequence for A15 was not as compatible as that of A1. The mRNA for CYP73A15 was shown to be rapidly induced by elicitor treatment of suspension cultured cells of French bean but induction was more transient than that of phenylalanine ammonia-lyase (PAL). In contrast, induction in cells undergoing xylogenesis was much more coordinate with PAL. The cloned cDNA may represent a cinnamate 4-hydroxylase isoform, whose expression is more related to differentiation than the responses to stress in which the majority of CYP73As cloned so far are involved. PMID- 10380797 TI - Splicing-independent processing of plant box C/D and box H/ACA small nucleolar RNAs. AB - Small nucleolar RNAs (snoRNAs) are involved in various aspects of ribosome biogenesis and rRNA maturation. Plants have a unique organisation of snoRNA genes where multiple, different genes are tightly clustered at a number of different loci. The maize gene clusters studied here include genes from both of the two major classes of snoRNAs (box C/D and box H/ACA) and are transcribed as a polycistronic pre-snoRNA transcript from an upstream promoter. In contrast to vertebrate and yeast intron-encoded snoRNAs, which are processed from debranched introns by exonuclease activity, the particular organisation of plant snoRNA genes suggests a different mode of expression and processing. Here we show that single and multiple plant snoRNAs can be processed from both non-intronic and intronic transcripts such that processing is splicing-independent and requires endonucleolytic activity. Processing of these different snoRNAs from the same polycistronic transcript suggests that the processing machineries needed by each class are not spatially separated in the nucleolus/nucleus. PMID- 10380798 TI - Cloning of a wheat puroindoline gene promoter by IPCR and analysis of promoter regions required for tissue-specific expression in transgenic rice seeds. AB - A genomic DNA fragment containing the 5'-upstream sequence and part of the open reading frame corresponding to Triticum aestivum puroindoline-b cDNA, was isolated by inverse PCR. Promoter fragments extending to -1068, -388, -210 or 124 upstream of the translation initiation ATG codon and the sequence coding for the first 13 amino acids of the puroindoline-b, were translationally fused to the uidA reporter gene encoding beta-glucuronidase and transferred to rice calli via particle bombardment-mediated transformation. The 1068 bp and 124 bp promoters were also transcriptionally fused to the uidA reporter gene. Out of the 196 plants regenerated from transformed rice calli, 118 plants set seeds. No GUS activity was detectable in the stems, roots, leaves or pollen of the transgenic rice which had integrated the puroindoline-b promoter or its deletions; GUS activity was detected only in seeds, except in those having integrated the 124 bp promoter. Within seeds, histological localisation showed GUS activity as being restricted to the endosperm, aleurone cells and pericarp cell layers; no GUS activity was detected in the embryonic axis. Analysis of 5' promoter deletions identified the region between -388 and -210 as essential for endosperm expression, and the region between -210 and -124 as essential for expression in the epithelium of the scutellum. No difference of expression was observed between the translational and transcriptional fusion genes. PMID- 10380799 TI - Characterization of ATDRG1, a member of a new class of GTP-binding proteins in plants. AB - We report the initial characterization of an Arabidopsis thaliana cDNA (atdrg1), a member of a new class of GTP-binding proteins (G-proteins) in plants. The predicted ATDRG1 protein contains all five structural motifs characteristic of the G-protein superfamily. Apart from these motifs, the amino acid sequence differs substantially from all known G-proteins except for a recently discovered new family named developmentally regulated G-proteins (DRGs). Sequences closely related to atdrg1 are found in species as distant as human (80% amino acid conservation), Drosophila (74%), yeast (77%) and Caenorhabditis elegans (77%). The remarkable evolutionary conservation of these proteins suggests an important, but as yet unclear role. Phylogenetic analysis of the available homologous sequences strongly suggests a diphyletic origin of the eukaryotic DRG proteins. Northern analysis shows high levels of atdrg1 mRNA in all Arabidopsis tissues studied, and homologues of atdrg1 are present throughout the plant kingdom. In situ hybridization reveals that atdrg1 is highly expressed in actively growing tissues and reproductive organs. Southern analysis indicates the presence of either one or two copies of atdrg1 in the Arabidopsis genome. Immunolocalization studies show that the protein is present in cytoplasmic vesicles found mainly in actively growing tissues suggesting a putative role for ATDRG1 in either the regulation of vesicle transport or the regulation of enzymes involved in storage protein processing. PMID- 10380800 TI - Unusual inheritance of evolutionarily-related double-stranded RNAs in interspecific hybrid between rice plants Oryza sativa and Oryza rufipogon. AB - Endogenous, 14 kb double-stranded RNAs (dsRNAs) have been found in two ecospecies of cultivated rice (temperate japonica rice and tropical japonica rice, Oryza sativa L.) and in wild rice (O. rufipogon, an ancestor of O. sativa). A comparison of the nucleotide and deduced amino acid sequences of the core regions of the RNA-dependent RNA polymerase domains found in these three dsRNAs suggested that these dsRNAs probably evolved independently within each host plant from a common ancestor. These dsRNAs were introduced into F1 hybrids by crossing cultivated rice and wild rice. Unusual cytoplasmic inheritance of these dsRNAs was observed in some F1 hybrids; the evolutionarily related dsRNAs were incompatible for each other, and the resident dsRNA of an egg cell from cultivated rice was excluded by the incoming dsRNA of a pollen cell from wild rice. Coexisting dsRNAs in the F1 hybrids segregated away from each other in the F2 plants. However, the total amount of these dsRNAs in the host cells remained constant (ca. 100 copies/cell). The stringent regulation of the dsRNA copy number may be responsible for their unusual inheritance. PMID- 10380801 TI - A distinct member of the basic (class I) chitinase gene family in potato is specifically expressed in epidermal cells. AB - We have isolated cDNA clones encoding class I chitinase (ChtC) from potato leaves which share a high degree of nucleotide and amino acid sequence similarity to other, previously described basic (class I) chitinases (ChtB) from potato. Despite this similarity, characteristic features distinguish ChtC from ChtB, including an extended proline-rich linker region between the hevein and catalytic domains and presence of a potential glycosylation site (NDT) in the deduced protein. These differences are in accordance with the properties of purified chitinase C which is glycosylated and hence has a higher molecular mass in comparison to chitinase B. In contrast to the coding sequences, the 3' untranslated regions of ChtC and ChtB exhibited a low degree of similarity, which allowed us to generate gene-specific probes to study the genomic organization and expression of both types of gene. Genomic DNA blots suggest that ChtC and ChtB are each encoded by one or two genes per haploid genome. RNA blot analysis showed that in healthy potato plants ChtC mRNA is most abundant in young leaves, the organs which also contain high levels of chitinase C. By contrast, ChtB mRNA abundance is highest in old leaves, which accumulate chitinase B. By in situ RNA hybridization with gene-specific probes we could demonstrate that ChtC mRNA in leaves is restricted to epidermal cells, whereas ChtB mRNA showed no distinct pattern of cell-type-specific localization. Infection of potato leaves with Phytophthora infestans, or treatment with fungal elicitor, ethylene, or wounding resulted in accumulation of both ChtC and ChtB mRNAs; however, for ChtC, in contrast to ChtB, no corresponding accumulation of the encoded protein could be detected, suggesting a post-transcriptional mechanism of regulation. Salicylic acid treatment did not induce accumulation of either mRNA. The possible functional implications of these findings for pathogen defence and developmental processes are discussed. PMID- 10380802 TI - ATS1 and ATS3: two novel embryo-specific genes in Arabidopsis thaliana. AB - A modified protocol for differential display of mRNA was used to identify and clone genes expressed in developing Arabidopsis thaliana seeds. Two novel embryo specific genes designated ATS1 and ATS3 (Arabidopsis thaliana seed gene) were identified. In situ hybridization showed that, spatially, ATS1 is expressed in a pattern similar to the Arabidopsis GEA1 gene and that ATS3 is expressed in a pattern similar to the Arabidopsis seed storage protein genes. Southern analysis of Arabidopsis genomic DNA indicated that ATS1 is a member of a small gene family and that ATS3 is present as a single copy in the diploid genome. Sequence analysis of both genes showed that ATS1 is similar to the rice EFA27 gene and that ATS3 is unique. Western analysis and light level immunocytochemistry using antisera raised against the putative ATS1 and ATS3 translation products verified that ATS1 and ATS3 proteins are seed-specific and accumulate in a spatial pattern similar to their respective transcripts. Taken together, these data show that ATS1 and ATS3 are novel embryo-specific genes in Arabidopsis. PMID- 10380803 TI - Quantitative chromosome map of the polyploid Saccharum spontaneum by multicolor fluorescence in situ hybridization and imaging methods. AB - Somatic chromosomes of a wild relative of sugarcane (Saccharum spontaneum L.) anther culture-derived clone (AP 85-361, 2n = 32) were identified and characterized by computer-aided imaging technology and molecular cytological methods. The presence of four satellite chromosomes and four nearly identical chromosome sets suggests that the clone is a tetrahaploid with the basic number x = 8. A quantitative chromosome map, or idiogram, was developed using image analysis of the condensation pattern (CP) at the prometaphase stage of somatic chromosomes. The 45S and 5S ribosomal RNA gene (rDNA) loci were simultaneously visualized by multi-color fluorescence in situ hybridization (McFISH) and precisely localized to the regions of 3p3.1 and 6q1.3 on the idiogram. The simultaneous visualization of two sets of four ribosomal RNA genes confirms tetraploidy of this clone. This conclusion is consistent with results of molecular marker mapping. The quantitative chromosome map produced will become the foundation for genome analyses based on chromosome identity and structure. Previously impossible identification of small chromosomes and untestable hypotheses about the polyploid nature of plants can now be settled with these two approaches of quantitative karyotyping and FISH. PMID- 10380804 TI - Genetic deletion of proteins resembling Type IV pilins in Synechocystis sp. PCC 6803: their role in binding or transfer of newly synthesized chlorophyll. AB - Upon non-denaturing gel electrophoresis of Synechocystis sp. PCC 6803 thylakoid extracts, a Type IV pilin-like protein encoded by open reading frame sll1694 was found in chlorophyll-containing bands. The Synechocystis sp. PCC 6803 genome also encodes two similar open reading frames, sll1695 and slr1456. Even though transcripts of sll1694 and slr1456 could be detected, deletion of the three open reading frames in systems with normal chlorophyll content had no effect. However, Sll1694 was found to affect the rate of chlorophyll synthesis and of the assembly of chlorophyll-binding proteins. In the sll1694/sll1695 deletion mutant in a PS I less/chlL- background, which is unable to synthesize chlorophyll in darkness, chlorophyll synthesis during the first hours of illumination after dark incubation was 30% slower than in the PS I-less/chlL- strain. Moreover, the biogenesis of chlorophyll-protein complexes with a 77K chlorophyll fluorescence emission maximum at 685 mm was delayed by several hours in this mutant whereas the rate of biogenesis of photosystem II was not significantly affected. Furthermore, results of non-denaturing gel electrophoresis indicated that a chlorophyll-binding complex formed during the early hours of chlorophyll synthesis was altered in stability and mobility upon deletion of the three open reading frames. We propose that the protein encoded by sll1694 is involved in, but is not absolutely required for, delivering chlorophyll to nascent photosystems and antennae. PMID- 10380805 TI - A cluster of five cell wall-associated receptor kinase genes, Wak1-5, are expressed in specific organs of Arabidopsis. AB - WAK1 (wall-associated kinase 1) is a cytoplasmic serine/threonine kinase that spans the plasma membrane and extends into the extracellular region to bind tightly to the cell wall. The Wak1 gene was mapped and found to lie in a tight cluster of five highly similar genes (Wak1-5) within a 30 kb region. All of the Wak genes encode a cytoplasmic serine/threonine protein kinase, a transmembrane domain, and an extracytoplasmic region with several epidermal growth factor (EGF) repeats. The extracellular regions also contain limited amino acid identities to the tenascin superfamily, collagen, or the neurexins. RNA blot analysis with gene specific probes revealed that Wak1, Wak3 and Wak5 are expressed primarily in leaves and stems of Arabidopsis. Wak4 mRNA is only detected in siliques, while Wak2 mRNA is found in high levels in leaves and stems, and in lower levels in flowers and siliques. A trace amount of Wak2 can also be detected in roots. Wak1 is induced by pathogen infection and salicylic acid or its analogue INA and is involved in the plant's response, and Wak2, Wak3 and Wak5 also can be greatly induced by salicylic acid or INA. The WAK proteins have the potential to serve as both linkers of the cell wall to the plasma membrane and as signaling molecules, and since Wak expression is organ-specific and the isoforms vary significantly in the cell wall associated domain this family of proteins may be involved in cell wall-plasma membrane interactions that direct fundamental processes in angiosperms. PMID- 10380806 TI - Sequences surrounding the transcription initiation site of the Arabidopsis enoyl acyl carrier protein reductase gene control seed expression in transgenic tobacco. AB - The NADH-specific enoyl-acyl carrier protein (ACP) reductase, which catalyses the last reducing step during the fatty acid biosynthesis cycle, is encoded in Arabidopsis thaliana encoded by a single housekeeping gene (ENR-A) which is differentially expressed during plant development. To identify elements involved in its tissue-specific transcriptional control, a fragment comprising the 1470 bp region directly upstream of the ATG start codon of the ENR-A gene was fused to the uidA (GUS) reporter gene and analysed in transgenic Nicotiana tabacum plants. GUS activity found during development of the transgenic plants was similar to endogenous ENR protein levels found in both tobacco and Arabidopsis plants, except for developing flowers. In floral tissue the promoter fragment showed very little activity in contrast to the relatively high level of endogenous ENR expression. Successive deletions from the 5' and 3' regions of the promoter fragment revealed the presence of at least three elements which control GUS expression in different stages of development in the transgenic tobacco plants. First, expression in young developing leaves required both the presence of sequences between -329 to -201 relative to the transcription start and part of the untranslated leader comprising the first intron. Second, root-specific GUS expression was still observed after deletion of the 5'-upstream sequences up to 19 bp of the transcription initiation site. Further, the additional removal of the intron from the untranslated leader increased root-specific expression by ca. 4- to 5-fold. Third, high expression in seeds was still observed with the minimal upstream promoter segment of 19 bp. This seed expression level was found to be independent of the presence or absence of the intron in the untranslated leader. Finally, 3' deletion of the leader sequence up to 17 bp of the transcription start greatly impaired GUS activity during all stages of plant development, suggesting that the deleted sequence of the leader either functions as an enhancer for transcription initiation or stabilizes the mRNA. PMID- 10380807 TI - Molecular analysis of a null mutant for pea (Pisum sativum L.) seed lipoxygenase 2. AB - A mutant line of Pisum fulvum was identified that lacked seed lipoxygenase-2 (LOX 2). The mutant phenotype was introgressed into a standard Pisum sativum cv. Birte to provide near-isogenic lines with or without seed LOX-2. Genetic analyses showed the mutation to behave as a single, recessive Mendelian gene. Northern and dot-blot analyses showed a large reduction in LOX-2 mRNA from developing seeds of the LOX-2-null mutant. A restriction fragment length polymorphism associated with the 5' end of the LOX-2 gene(s) co-segregated with the null phenotype, indicating that the reduction of LOX-2 mRNA was neither a consequence of deletion of the LOX genes nor a consequence of the action of a genetically distant regulatory gene. Analysis of the 5'-flanking sequences of LOX-2 genes from Birte and the near isogenic LOX-2-null mutant revealed a number of insertions, deletions and substitutions within the promoter from the LOX-2-null mutant that could be responsible for the null phenotype. Incubation of crude seed LOX preparations from Birte and the LOX-2-null mutant showed that the latter generated relatively less 13-hydroperoxides and also produced relatively more hydroxy- and ketoacid compounds that have implications for the fresh-frozen pea industry. PMID- 10380808 TI - A promoter from sugarcane bacilliform badnavirus drives transgene expression in banana and other monocot and dicot plants. AB - A 1369 bp DNA fragment (Sc) was isolated from a full-length clone of sugarcane bacilliform badnavirus (ScBV) and was shown to have promoter activity in transient expression assays using monocot (banana, maize, millet and sorghum) and dicot plant species (tobacco, sunflower, canola and Nicotiana benthamiana). This promoter was also tested for stable expression in transgenic banana and tobacco plants. These experiments showed that this promoter could drive high-level expression of the beta-glucuronidase (GUS) reporter gene in most plant cells. The expression level was comparable to the maize ubiquitin promoter in standardised transient assays in maize. In transgenic banana plants the expression levels were variable for different transgenic lines but was generally comparable with the activities of both the maize ubiquitin promoter and the enhanced cauliflower mosaic virus (CaMV) 35S promoter. The Sc promoter appears to express in a near constitutive manner in transgenic banana and tobacco plants. The promoter from sugarcane bacilliform virus represents a useful tool for the high-level expression of foreign genes in both monocot and dicot transgenic plants that could be used similarly to the CaMV 35S or maize polyubiquitin promoter. PMID- 10380809 TI - Gene encoding polygalacturonase inhibitor in apple fruit is developmentally regulated and activated by wounding and fungal infection. AB - A cDNA encoding polygalacturonase-inhibiting protein (PGIP) from mature apple fruit has been cloned and characterized. The open reading frame encodes a polypeptide of 330 amino acids, in which 24 amino acids at the N-terminus comprise the signal peptide. Apple PGIP contains 10 imperfect leucine-rich repeat sequence motifs averaging 24 amino acids in length. In addition to the 1.3 kb PGIP transcript, the cloned cDNA also hybridized to RNA molecules with sizes of 3.2 and 5.0 kb. Genomic DNA analysis revealed that the apple PGIP probably belongs to a small family of genes. PGIP transcript levels varied in fruit collected at different maturities, suggesting the gene is developmentally regulated. Very high PGIP transcript levels were detected in decayed areas and the tissue adjacent to the inoculation sites of Penicillium expansum and Botrytis cinerea. However, no increase in the amount of PGIP transcript in tissue distant from the decayed region was observed. Wounding on fruit also induced PGIP gene expression but to a much lessser extent when compared with decayed areas. After storage at 0 degrees C for 1 month, the abundance of PGIP transcript in ripe fruit was substantially increased. The PGIP gene in immature and ripe fruit was rapidly up-regulated by fungal infections, while in stored fruit the induction was very limited and concurred with an increase of fruit susceptibility to fungal colonization. Since PGIP gene expression is regulated by fruit development and responds to wounding, fungal infection and cold storage, these observations suggest that apple PGIP may have multiple roles during fruit development and stress response. PMID- 10380810 TI - Markers for hypersensitive response and senescence show distinct patterns of expression. AB - Controlled cellular suicide is an important process that can be observed in various organs during plant development. From the generation of proper sexual organs in monoecious plants to the hypersensitive response (HR) that occurs during incompatible pathogen interactions, programmed cell death (PCD) can be readily observed. Although several biochemical and morphological parameters have been described for various types of cell death in plants, the relationships existing between those different types of PCD events remain unclear. In this work, we set out to examine if two early molecular markers of HR cell death (HIN1 and HSR203J) as well as a senescence marker (SAG12) are coordinately induced during these processes. Our result indicates that although there is evidence of some cross-talk between both cell death pathways, spatial and temporal characteristics of activation for these markers during hypersensitive response and senescence are distinct. These observations indicate that these markers are relatively specific for different cell death programs. Interestingly, they also revealed that a senescence-like process seems to be triggered at the periphery of the HR necrotic lesion. This suggests that cells committed to die during the HR might release a signal able to induce senescence in the neighboring cells. This phenomenon could correspond to the establishment of a second barrier against pathogens. Lastly, we used those cell death markers to better characterize cell death induced by copper and we showed that this abiotic induced cell death presents similarities with HR cell death. PMID- 10380811 TI - Enzymatic activity and gene expression under water stress of phospholipase D in two cultivars of Vigna unguiculata L. Walp. differing in drought tolerance. AB - Phospholipase D, a major lipid-degrading enzyme in plants, was studied in two cultivars of Vigna unguiculata L.Walp, differing in their tolerance to drought (cv. EPACE-1, drought-tolerant, and cv. 1183, drought-susceptible). Enzymatic activities, measured with 14C-PC as substrate, increased when plants were submitted to water stress, the increase being much higher in the drought sensitive cultivar. A 2911 bp cDNA encoding a putative phospholipase D (VuPLD1) was isolated from a cDNA library prepared from V. unguiculata leaves. The deduced amino acid sequence (809 residues) shows 85.5% identity and 91.3% similarity to that of PLD from Ricinus communis. The expression of the VuPLD1 gene in the leaves is differently modulated by water deficit, depending on the intensity of stress and the tolerance or sensitivity of the plants. In the drought-susceptible V. unguiculata cv. 1183, it readily increased under water stress, reaching maximum values at mild water deficit (-1.5 MPa). In the drought-tolerant cv. EPACE-1, VuPLD1 mRNA remained low throughout the whole drought treatment. Dehydration of leaves led to a dramatic increase in transcript level in both cultivars. Changes in protein amounts semi-quantified by immunoblotting correlated well with variations in transcript steady-state level. Taken together, these results showed that phospholipase D in cowpea plants is essentially regulated at the transcriptional level, and that gene expression is strongly stimulated even by moderate water deficit in the drought-sensitive plant. On the contrary, the drought-tolerant plant presents a remarkable stability of PLD gene expression in conditions of water stress. PMID- 10380812 TI - Engineering seed dormancy by the modification of zeaxanthin epoxidase gene expression. AB - Abscisic acid (ABA) is a plant hormone synthesized during seed development that is involved in the induction of seed dormancy. Delayed germination due to seed dormancy allows long-term seed survival in soil but is generally undesirable in crop species. Freshly harvested seeds of wild-type Nicotiana plumbaginifolia plants exhibit a clear primary dormancy that results in delayed germination, the degree of primary dormancy being influenced by environmental culture conditions of the mother plant. In contrast, seeds, obtained either from ABA-deficient mutant aba2-s1 plants directly or aba2-s1 plants grafted onto wild-type plant stocks, exhibited rapid germination under all conditions irrespective of the mother plant culture conditions. The ABA biosynthesis gene ABA2 of N. plumbaginifolia, encoding zeaxanthin epoxidase, was placed under the control of the constitutive 35S promoter. Transgenic plants overexpressing ABA2 mRNA exhibited delayed germination and increased ABA levels in mature seeds. Expression of an antisense ABA2 mRNA, however, resulted in rapid seed germination and in a reduction of ABA abundance in transgenic seeds. It appears possible, therefore, that seed dormancy can be controlled in this Nicotiana model species by the manipulation of ABA levels. PMID- 10380813 TI - In plants a putative isovaleryl-CoA-dehydrogenase is located in mitochondria. AB - In plants the degradation pathways of branched-chain amino acids have remained somewhat unclear with respect to both their biochemistry and their intracellular location. While biochemical evidence has localized some of the catabolic enzymes in peroxisomes/glyoxysomes, others cofractionate with mitochondria. We have now identified a candidate protein and corresponding cDNA for an enzyme of the leucine catabolic pathway, the isovaleryl-CoA-dehydrogenase (IVD). This polypeptide is a member of the acyl-CoA-dehydrogenase (ACDH) family and is encoded in the nuclear genome of Arabidopsis thaliana. Expression of the putative IVD gene in pea seedlings is documented by western blot analyses with an antibody against the mammalian IVD. Subcellular fractionation identifies the putative IVD enzyme in the mitochondrion. This localization suggests that in plants mitochondria contain at least part of the branched-chain amino acid degradation pathway(s). PMID- 10380814 TI - Molecular characterization of two lipoxygenases from barley. AB - Two full-length lipoxygenase cDNA sequences (LoxB and LoxC) from barley (Hordeum distichum cv. L. Triumph) are described. The cDNAs share high homology with the barley LoxA cDNA. Southern blotting experiments indicate single copy numbers of the three lipoxygenase genes. RFLP mapping revealed the presence of single lipoxygenase loci. LoxA and LoxB map on chromosome 4 and LoxC on chromosome 7. Two isoenzymes, LOX1 and LOX2, have been purified previously from germinating barley and characterized. LOX1 is encoded by LoxA, while LOX2 is encoded by LoxC. The product related to the third cDNA (loxB) has not been identified so far, suggesting a low protein abundance for the corresponding isoform in barley. Transcripts corresponding with these LOX genes are predominantly observed in grain and in seedling, whereas transcripts corresponding to LoxB and LoxC are also observed in mature vegetative tissue. No lipoxygenase mRNA could be detected in aleurone layer of germinating grain. No significant differences in lipoxygenase mRNA levels were observed in developing grains grown under dormant or non-dormant conditions, suggesting that LOX is not directly involved in induction of grain dormancy. PMID- 10380816 TI - Survival of cancer patients in Finland, 1955-1994. PMID- 10380817 TI - The 'Vth Scandinavian Meeting on Radioimmuno-targeting', Umea, Sweden, February 25-27, 1998. PMID- 10380815 TI - The promoter of the strictosidine synthase gene from periwinkle confers elicitor inducible expression in transgenic tobacco and binds nuclear factors GT-1 and GBF. AB - Strictosidine synthase (STR) is a key enzyme in the biosynthesis of terpenoid indole alkaloids. This class of secondary metabolites harbours several pharmaceutically important compounds used, among other applications, in cancer treatment. Terpenoid indole alkaloid biosynthesis and expression of biosynthetic genes including Str1 is induced by fungal elicitors. To identify elicitor responsive regulatory promoter elements and trans-acting factors, the single-copy Str1 gene was isolated from the subtropical plant species Catharanthus roseus (Madagascar periwinkle). Str1 upstream sequences conferred elicitor-responsive expression to the beta-glucuronidase (gusA) reporter gene in transgenic tobacco plants. Main enhancer sequences within the Str1 promoter region studied were shown to be located between -339 and -145. This region and two other regions of the promoter bound the tobacco nuclear protein factor GT-1. A G-box located around position -105 bound nuclear and cloned G-box-binding factors (GBFs). A mutation that knocked out GBF binding had no measurable effect on expression, which indicates that the G-box is not essential for the elicitor responsiveness of the Str1 promoter. No obvious homologies with promoter elements identified in other elicitor-responsive genes were observed, suggesting that the Str1 gene may depend on novel regulatory mechanisms. PMID- 10380818 TI - Cancer patient survival--patterns, comparisons, trends--a population-based Cancer Registry study in Finland. AB - The effects of primary site, sex, age, stage and histological type on cancer patient survival were analysed on the basis of the population-based material of the Finnish Cancer Registry from 1985 to 1994. In addition, trends in survival were constructed for the period 1955-1994. Detailed site-specific data are published as Supplement 12 to Vol. 38 of Acta Oncologica. Within a given site, the survival differences by gender were not large. However, because of different site distributions, the average prognosis for female patients, all sites taken together, was superior to that of males: the 5-year relative survival rates (RSR) were 58% and 43%, respectively. In general, older patients had a poorer outcome compared with younger patients (partly because of different stage and histology distributions). Stage was a strong determinant of patient survival. In some cancers with a poor average prognosis the 5-year RSR for localized tumours was reasonable, e.g. 61% for stomach cancer, males, 34% for gallbladder cancer, females, and 29% for lung cancer, males. Most of the survival rates clearly increased over time. In addition to improvements in cancer treatment, changes over time in several other factors affect the trends, such as changes in the stage distribution (early diagnosis as a result of health education, improved diagnostic methods, screening, etc.) and in the composition of the patient material because of changing definitions of cancer (e.g. papilloma versus papillary carcinoma of the bladder, occult carcinoma of the thyroid, and early prostate cancer). The large Cancer Registry material (466,000 patients) enabled accurate estimates of the survival rates of cancer patients in Finland. These rates reflect the effectiveness of the healthcare system as a whole and are useful for planning and evaluation purposes. However, the estimated survival rates are based on grouped data, and cannot be directly applied for predicting the prognoses of individual patients, although they can be used as guidelines. PMID- 10380819 TI - The rationale for early diagnosis of cancer--the example of breast cancer. AB - The main advantage of early diagnosis of cancer is the reduction of tumor size at initial treatment and thereby an increase in the proportion of patients without distant dissemination. This benefit is illustrated by the example of breast cancer. A model of its natural history was built using data extracted from the files of over 4000 patients followed in the same institution for 20 to 35 years. The model was used to quantify the impact of tumor size, histologic grade, and lymph node involvement on the probability of distant spread. The relationships were found to be highly significant. The model also and unexpectedly revealed that the tumors progress while they grow; avoiding histologic progression is therefore another advantage of early diagnosis. The model showed that residual tumor can be a nidus for distant dissemination, and that consistency between the prediction of the model and the results of post-operative radiotherapy is satisfactory. Conversely to what is often stated, the benefits of post-operative radiotherapy appear to be greater for small tumors, even in the absence of lymph node involvement, than for large ones. The model could be used to help improve screening strategies, but more data are required, in particular for the young age range. PMID- 10380820 TI - Ethical management of hereditary cancer information. AB - Genetic diagnosis yields information that is highly relevant for both the patient and the genetic relatives of the patient. In this article two ethical problems are discussed. Under what conditions should hereditary cancer information be given to a relative? It is suggested that in order to answer this question, three factors have to be considered and a balance struck: the seriousness of the condition, the existence of treatment or prevention and the reliability of the diagnosis. The second issue discussed in the article relates to the psychosocial effects of giving hereditary cancer information. It is argued that ethical management of clinical practice requires that further attention must be given to the psychosocial effects on both the individual and the family. PMID- 10380821 TI - Immunogenetic properties of genes. AB - A most surprising finding is that when injected into animal tissue naked DNA is taken up and expressed by cells with great efficiency. The DNA is given as a plasmid that includes a promoter and an enhancer. When inoculated, the DNA is not stably integrated within the chromosomal DNA, but persists as extrachromosomal nuclear episomes. Both purified DNA and RNA have been shown to be expressed in somatic cells. It is now well established that injection of DNA by many routes may result in in vivo expression and that the proteins become immunogenic. DNA vaccines seem to induce efficient and complete immune responses. The immunizations produce long-term humoral and cellular responses, qualitatively similar to live attenuated vaccines but without the safety hazards of infectious agents. Recently, the first human DNA vaccinations were reported. PMID- 10380822 TI - Conjugate chemistry and cellular processing of EGF-dextran. AB - Conjugates with specific binding to the epidermal growth factor receptor, EGFR, of interest for radionuclide based imaging and therapy were prepared using mouse epidermal growth factor, mEGF, and dextran. In one type of conjugate, mEGF was coupled to dextran by reductive amination in which the free amino group on the mEGF N-terminal reacted with the aldehyde group on the reductive end of dextran. The end-end coupled conjugate could be further activated by the cyanopyridinium agent CDAP, thereby introducing tyrosines to the dextran part. In the other type of conjugate, the cyanylating procedure using CDAP was applied, first to activate dextran and then allowing for the amino terminus of mEGF to randomly attach to the dextran. In the latter case, radionuclide-labelled tyrosines or glycines could be added in the same conjugation step. All types of mEGF-dextran conjugates had EGFR-specific binding since the binding could be displaced by an excess of non-radioactive mEGF. The conjugates were to a large extent internalized in the test cells and the associated radioactivity was retained intracellularly for different times depending on both the type of cells and conjugate applied. Different intracellular 'traffic routes' for the radionuclides are discussed as well as applications for both imaging and therapy. PMID- 10380823 TI - Biokinetics of the monoclonal antibodies MOv 18, OV 185 and OV 197 labelled with 125I according to the m-MeATE method or the Iodogen method in nude mice with ovarian cancer xenografts. AB - The biodistribution of the radiolabelled monoclonal antibodies MOv18, OV185 and OV197 in nude mice with subcutaneous tumours of the human ovarian cancer cell line OVCAR3 was investigated. The early uptake of MOv18 (1-24 h) and the uptake in relation to tumour size were also studied. The antibodies were labelled with 125I according to the Iodogen method or the m-MeATE method, the latter also being suitable for labelling with 211Astatine. The tumour/blood ratio and the localization index for Mov 18 72 h after antibody injection were 2.21 +/- 0.25 and 4.62 +/- 1.27, respectively. This is significantly higher than for the other two specific antibodies. The early tumour uptake of MOv18 was low with a tumour/blood ratio of 0.23 +/- 0.04 after 6 h, and the uptake was higher in small tumours. The two labelling methods were found to be equivalent. We conclude that MOv18 labelled according the m-MeATE method should be suitable for further therapeutic studies with 211Astatine in nude mice. PMID- 10380824 TI - Stability and immunoreactivity of the monoclonal anticytokeratin antibody TS1 after different degrees of iodination. AB - The immunoreactivity, stability and in vivo kinetics of an anticytokeratin 8 monoclonal antibody, TS1, were investigated following different degrees of labeling with 125I (0.2, 1 and 2-3 125I/TS1 MAb). By testing with ELISA, it was demonstrated that a high degree of iodination, i.e. > 2 125I/TS1, caused a rapid decrease in immunoreactivity to almost zero within 10 days. Furthermore, a complete degradation to low molecular weight fragments and free iodine was seen, as shown by SDS PAGE and autoradiography. The differently labeled radionuclide conjugates were injected into nude mice inoculated with HeLa Hep2 cells and tumor doses (estimated by MIRD formalism), tumor:non-tumor dose ratios, % I.D./gram tissue, Gy/MBq and in vivo kinetics of the differently labeled MAbs were determined. Despite the in vitro instability of the highest iodinated radionuclide conjugate, it was possible to deliver high doses to the tumors if the conjugate was injected into the animal immediately after completion of the iodination procedure. Increases from 1.4 Gy to 15.2 Gy delivered tumor dose were obtained with a tenfold increase in the specific activity, without alterations in the tumor:non-tumor tissue dose ratios. There is room for significant improvements in efficacy at radioimmunotherapy, which can be gained by optimizing the degree of iodination. For therapeutical applications a high degree of iodination may be an advantage. PMID- 10380825 TI - Positron emission tomography and radioimmunotargeting--general aspects. AB - To optimize radioimmunotherapy, in vivo information on individual patients, such as radionuclide uptake, kinetics, metabolic patterns and optimal administration methods, is important. An overriding problem is to determine accurately the absorbed dose in the target organ as well as critical organs. Positron Emission Tomography (PET) is a superior technique to quantify regional kinetics in vivo with a spatial resolution better than 1 cm3 and a temporal resolution better than 10 s. However, target molecules often have distribution times of several hours to days. Conventional PET nuclides are not applicable and alternative positron emitting nuclides with matching half-lives and with suitable labelling properties are thus necessary. Over many years we have systematically developed convenient production methods and labelling techniques of suitable positron nuclides, such as 110In(T(1/2) = 1.15 h), 86Y(T(1/2) = 14 h), 76Br(T(1/2) = 16 h) and 124I(T(1/2) = 4 days). 'Dose planning' can be done, for example, with 86Y- or 124I-labelled ligands before therapy, and 90Y- and 131I-labelled analogues and double-labelling, e.g. with a 86Y/90Y-labelled ligand, can be used to determine the true radioactivity integral from a pure beta-emitting nuclide. The usefulness of these techniques was demonstrated in animal and patient studies by halogen labelled MAbs and EGF-dextran conjugates and peptides chelated with metal ions. PMID- 10380826 TI - Positron emission tomography and radioimmunotargeting--aspects of quantification and dosimetry. AB - Positron emission tomography (PET) is a medical imaging tool with high resolution and good quantitative properties, which makes it suitable for in vivo quantification of radioimmunotargeting agents. Most radionuclides used in radioimmunotherapy have positron-emitting analogues, which can be used for PET imaging, and this opens the possibility of performing dosimetry with PET. These isotopes, however, often emit gamma radiation and high-energy positrons in their decay, influencing the imaging properties of PET. Spatial resolution, reconstructed background and line source recovery for a number of non-pure positron emitters were investigated and compared with the imaging properties of 18F. PET imaging properties did not degrade severely for these non-pure positron emitters, but caution has to be applied when doing quantitative measurements. To assess the possibility of conducting PET studies during therapy, by combining, for example, a small amount of 124I with 131I, the influence of the presence of large amounts of gamma radiation on PET count rate characteristics was studied. The results of these studies were related to the necessary amounts of radioactivity needed for treatment of post-operative remains of glioma. The results indicate that the count rate capabilities of 2D PET permit PET studies for dose evaluation during radioimmunotherapy. PMID- 10380828 TI - Methods for estimating uptake and absorbed dose in tumours from 125I labelled monoclonal antibodies, based on scintigraphic imaging of mice. AB - Monoclonal antibodies for radioimmunotargeting are often tested in tumour bearing nude mice. In vivo determination of the uptake of the monoclonal antibody in the tumour requires quantitative scintigraphy, and this in turn requires an adequate method for subtraction of radiation from the normal tissue. For this reason, two different methods for background subtraction were evaluated, a contralateral background region of interest or an irregular one, surrounding the tumour. A pinhole collimator was used for the scintigraphy and the monoclonal antibodies were labelled with 125I. Furthermore, a method was developed for estimation of the mean absorbed dose in the tumour from these repeated quantitative scintigraphic measurements. This requires that the tumour mass can be accurately estimated in vivo. Finally, the results were compared with in vitro measurements of the uptake. PMID- 10380827 TI - 131I radioconjugated antibodies for the locoregional radioimmunotherapy of high grade malignant glioma--phase I and II study. AB - Locoregional radioimmunotherapy (LR-RIT) was administered to 111 patients (20 were recruited in a phase I and 91 in a phase II study) with malignant gliomas: 1 patient with oligodendroglioma, 7 patients with anaplastic oligodendroglioma, 2 with grade II astrocytoma, 10 with anaplastic astrocytoma and 91 with glioblastoma, amounting to 58 newly diagnosed and 53 recurrent tumours. The 131I labelled monoclonal antibodies BC-2 and BC-4 were used in order to recognize stromal and intracellular glycoprotein tenascin, an antigen present particularly in glioblastoma. The patients were enrolled between February 1990 and December 1997 after conventional therapy. The radiopharmaceutical was injected directly into the tumour site. Sequential scintigraphies demonstrated a high and enduring uptake in the tumour. The mean irradiation dose in the tumour was 300 Gy per cycle. In the group of 74 phase II glioblastoma patients the clinical responses were as follows: 10 patients with stable disease (SD), 9 with partial responses (PR), 23 with no evidence of disease (NED) and 1 patient with complete response (CR). The median survival was 19 months. The response rate (CR + PR + NED) was 17.8% for those patients with bulky lesions, with a median survival of 17 months, but 66.6% for patients with small lesions, with a median survival of 25 months. Better outcomes were recorded in cases with less aggressive diseases: oligodendroglioma, anaplastic oligodendroglioma and anaplastic astrocytoma. We conclude that fractionated LR-RIT can be safely performed, with promising results especially in patients with minimal disease. PMID- 10380829 TI - Cluster models in cellular level electron dose calculations. AB - A program for calculating absorbed dose was developed for radioimmunotherapy (RIT) purposes. It was used to determine the difference in the therapeutic effect of (111)In electrons when using a close-packed cubic geometry and a cell cluster model developed in this project. Our cluster model piles the cells individually. The cells were modelled as spheres of diameters of 12 (tumour) and 30 (healthy) microm. Both models were used to generate clusters with spherical tumours inside healthy tissue. The program uses Monte Carlo-based dose kernels. The radiation spectra were calculated from the Auger and x-ray transition strengths and fluorescence yields of (111)In. The results show the importance of the cluster model in cellular level dose calculations. Near the tumour/healthy tissue interface in particular, the doses differ because of geometrical differences. In the case of a small cluster with tumour and total diameters of 30 and 150 microm, the ratio of the therapeutic effects is 20. PMID- 10380830 TI - Treatment with high dose [(111)In-DTPA-D-PHE1]-octreotide in patients with neuroendocrine tumors--evaluation of therapeutic and toxic effects. AB - Carcinoid tumors and endocrine pancreatic tumors often express somatostatin receptors (sst). Tumor spread may be visualized by sst scintigraphy using [(111)In-DTPA-D-Phe1]-octreotide. In this study, tumor targeting therapy with [(111)In-DTPA-D-Phe1]-octreotide at high doses (6 GBq every third week) was used to treat patients with sst-expressing tumors. Five patients entered the protocol and three were evaluable for response, while all could be evaluated for toxicity. Two patient responded with a significant reduction in tumor markers (> 50%). The third patient showed increasing levels of tumor markers. Side effects were expressed as depression of bone-marrow function. In one patient a grade 4 reduction in platelet count was observed requiring several thrombocyte transfusions. In another two patients platelet counts decreased significantly. We conclude that treatment with [(111)In-DTPA-D-Phe1]-octreotide can be used in patients with neuroendocrine tumors but blood parameters have to be carefully monitored to avoid severe side effects. PMID- 10380831 TI - Intravesical administration of radiolabelled tumour-associated monoclonal antibody in bladder cancer. AB - Considerable progress has been made over the past decade in the use of tumour associated monoclonal antibodies (mAbs) as carriers of cytotoxic agents in the management of several malignancies. In the present study we investigated the tumour localization and biodistribution of the mAb HMFG1 to bladder cancer following intravesical administration. HMFG1, which has been raised against the polymorphic epithelial mucin (PEM), was labelled with 125I and administered intravesically 2 h and 24 h before cystoscopy. During cystoscopy, biopsies were taken from the normal bladder and the malignant lesion. Tissue samples were tested for HMFG1 immunostaining and for radioactivity with the use of a gamma counter. The mean ratio of tumour to normal uptake for 125I-HMFG1 was 5.81 +/- 1.23, at 2 h and 2.17 +/- 0.42 at 24 h. No radioactivity was detected in the blood. We conclude that HMFG1 could be used intravesically for the successful delivery of a cytotoxic agent. PMID- 10380832 TI - In vivo cellular distribution and endocytosis of the somatostatin receptor-ligand complex. AB - Radioactive tumor targeting agents are highly interesting and for treatment of neuroendocrine tumors expressing somatostatin receptors, radiolabeled somatostatin analogues (including [(111)In-DTPA-D-Phe1]-octreotide) has been tried in a small number of patients with encouraging results. To increase our knowledge about the in vivo processing of administered [(111)In-DTPA-D-Phe1] octreotide we have examined tumor and normal tissue material from a patient with a midgut carcinoid tumor. By ultrastructural autoradiography, silver grains indicating the presence of [(111)In-DTPA-D-Phe1]-octreotide could be identified within tumor cells, both in the primary tumor and in the mesenteric metastases. Silver grains were also found in leukocytes and in blood vessels. However, normal enterocytes did not show any specific radioligand uptake. This study indicates that the binding and endocytosis of [(111)In-DTPA-D-Phe1]-octreotide is a specific process that takes place in cells expressing somatostatin receptors. However, the importance of the number of somatostatin receptors and subtypes expressed will have to be further studied. PMID- 10380833 TI - Psychological distress and rheumatic disease. AB - Psychological factors influence the results of self-reports of pain, function and global severity in questionnaires such as the HAQ, SF-36 and the WOMAC. Persons with psychological distress use more resources, including medications, and have greater rates of work disability and joint surgery. Psychological status is influenced only very slightly by disease severity and tends to remain relatively constant over the course of the rheumatic disease. The psychological status of patients with differing rheumatic diseases is similar, and patients with rheumatoid arthritis do not have special psychological problems or psychological characteristics. PMID- 10380834 TI - Psychological factors in chronic rheumatic diseases--a review. The case of rheumatoid arthritis, current research and some problems. AB - An overview of studies relating psychological factors to perceived well-being among Rheumatoid Arthritis (RA) patients is presented. Most attention has been devoted to the perception of control, coping and the effects of cognitive distortions. The introduction of these constructs have advanced the understanding of psychological distress among RA patients, although they explain only a smaller part. One reason could be that they give an oversimplified picture of adjustment processes in chronic and disabling diseases. This may partly be because their development are based either on studies of depression in a psychiatric sense or on how healthy subjects manage stressful events in daily life. The results therefore have limited relevance for adjustment to a life-long, chronic illness like RA. Recent research has also suggested that personality dispositions, especially neuroticism, play a substantial role in all types of subjective experiences. Finally, some issues for future research are discussed. PMID- 10380835 TI - Muscle strength characteristics and central bone mineral density in women with recent onset rheumatoid arthritis compared with healthy controls. AB - Muscle strength and bone mineral density (BMD) at the lumbar spine (BMDspine) and femoral neck (BMDfem) were determined in 20 healthy women and in 20 women with recent onset rheumatoid arthritis (RA). The mean duration of articular symptoms of the patients was eleven months and none of them had used glucocorticoids or disease modifying antirheumatic drugs. BMDs were measured by dual x-ray absorptiometry (DXA). Knee extension, trunk extension, and flexion as well as grip strength were measured with David 200 and Digitest dynamometers. BMDspine (1.17 g/cm2 and 1.20 g/cm2) and BMDfem (0.98 g/cm2 and 0.96 g/cm2) between the women with early RA and healthy women did not differ. However, knee extension strength was 46%, grip strength 31%, trunk extension strength 14% and overall muscle strength index 29% lower in RA women (p < 0.020-0.001) than in healthy subjects. Femoral neck BMD correlated statistically significantly with knee extension strength and muscle strength index in both groups and with trunk extension and flexion strength as well as rapid force development in RA women. The data indicates that the loss of muscle strength is clearly visible during the first months of disease but the significant bone loss at central bone regions develops later. PMID- 10380836 TI - The prevention of corticosteroid-induced bone loss with intermittent cyclical etidronate. AB - A prospective, randomised, double-blind, placebo controlled primary prevention trial was undertaken in 28 patients commencing low to moderate doses of corticosteroids for the first time. Patients were randomised to intermittent cyclical etidronate (400 mg daily for 2 weeks) and calcium (500 mg daily for 11 weeks) or intermittent cyclical placebo with calcium. After 52 weeks of treatment, lumbar spine BMD increased by 1.8% in the etidronate group, while it decreased by 3.7% in the placebo group. The differences in bone loss rate were statistically significant (p<0.01) at both 6 and 12 months. Similar trends were observed at the proximal femur, but differences were not statistically significant. These results suggest that intermittent cyclical etidronate therapy is effective in the primary prevention of corticosteroid-induced bone loss at the lumbar spine. PMID- 10380837 TI - Adverse drug reactions in Sjogren's syndrome. Frequent allergic reactions and a specific trimethoprim-associated systemic reaction. AB - Trimethoprim-associated systemic reactions, including aseptic meningitis, have been reported to be very rare adverse drug reactions. Patients with Sjogren's syndrome have been overrepresented, but no epidemiological surveys of the reaction have been conducted. To study the overall frequency of adverse drug reactions, and especially trimethoprim-associated reactions, we interviewed 85 primary Sjogren's syndrome patients and compared the results with those of 45 similarly interviewed osteoarthritis patients. Antimicrobial allergy was more common among Sjogren's syndrome patients than in osteoarthritis patients (46% vs. 27%). Eleven Sjogren's syndrome patients (13%), but no osteoarthritis patient, had experienced at least a partial, non-allergic systemic reaction with trimethoprim. Of them five (6%) had had a full-blown systemic reaction including both chills/fever and headache/backache and at least one of the following: malaise, vomiting, dizziness, confusion or meningeal irritation. Our findings confirm that allergic reactions to antimicrobials are frequent in Sjogren's syndrome. In addition to allergic reactions Sjogren's syndrome patients are prone to a specific trimethoprim-associated systemic reaction. This should be remembered when prescribing antimicrobials. PMID- 10380838 TI - Lower disease activity and disability in Swedish patients with rheumatoid arthritis in 1995 compared with 1978. AB - The aim of this study was to evaluate differences in disease activity, disability, and medical treatment in consecutive patients with rheumatoid arthritis seen at the outpatient clinics in Malmo, in 1978 (n = 148) and 1995 (n = 164). The groups were similar with regard to age, gender, disease duration, and the proportion having had hip or knee replacement surgery. The patients in 1995 had lower values for CRP (p<0.001), Ritchie Articular Index (mean values: 5.5 vs. 9.9, p<0.001), and Steinbrocker functional class index (mean values: 1.96 vs. 2.16, p<0.001) than the 1978 group. The 1995 patient group was also more extensively treated with DMARD:s (68 vs. 51%, p<0.01) and glucocorticosteroids (23 vs. 12%, p<0.02) and had historically been treated with almost twice as many DMARD:s (2.7 vs. 1.5, p<0.001). Similar findings regarding disease activity and disability were made when restricting the analysis to subgroups of patients that were seropositive or had a shorter disease duration (< 5 yrs). The lower disease severity in the 1995 group may be secondary to a more active medical treatment, although other possibilities such as differences in selection and secular changes in disease severity unrelated to medication cannot be excluded. PMID- 10380839 TI - Relationship between CD4+/CD8+ T cell ratio and T cell activation in systemic lupus erythematosus. AB - We investigated the relationship between the ratio of CD4+ to CD8+ T cells (CD4/CD8 ratio) and T cell activation, indicated by human leukocyte antigen (HLA) DR expression, in patients with systemic lupus erythematosus (SLE). We found that the ratio was decreased in SLE patients and that this was significantly related to expression of HLA-DR by CD8+ (but not CD4+) T cells. These findings may assist in understanding the pathogenesis of SLE. In some SLE patients, the CD4/CD8 ratio and HLA-DR expression may be good indicators of therapeutic efficacy. PMID- 10380840 TI - The distribution of YKL-40 in osteoarthritic and normal human articular cartilage. AB - YKL-40, also called human cartilage glycoprotein-39, is a major secretory protein of human chondrocytes in cell culture. YKL-40 mRNA is expressed by cartilage from patients with rheumatoid arthritis, but is not detectable in normal human cartilage. The aim was to investigate the distribution of YKL-40 in osteoarthritic (n=9) and macroscopically normal (n=5) human articular cartilage, collected from 12 pre-selected areas of the femoral head, to discover a potential role for YKL-40 in cartilage remodelling in osteoarthritis. Immunohistochemical analysis showed that YKL-40 staining was found in chondrocytes of osteoarthritic cartilage mainly in the superficial and middle zone of the cartilage rather than the deep zone. There was a tendency for high number of YKL-40 positive chondrocytes in areas of the femoral head with a considerable biomechanical load. The number of chondrocytes with a positive staining for YKL-40 was in general low in normal cartilage. The present findings, together with previous observations, suggests that YKL-40 may be of importance in cartilage remodelling/degradation of osteoarthritic joints. PMID- 10380842 TI - Neural compressive symptoms appearing during steroid treatment in a patient with intracranial lipoma. AB - Intracranial lipoma is a rare condition, and it is usually asymptomatic. We describe a 67 year old woman who developed blurred vision, diplopia, left sided oculomotor palsy, and ipsilateral ptosis during steroid treatment for giant cell arteritis. These symptoms were considered to be associated with aggressive giant cell arteritis, and the steroid dose was raised. Surprisingly, the symptoms increased, and further examination revealed an intracranial lipoma situated in the Meckel's cave. During tapering of the steroids her symptoms gradually improved. This is the first report demonstrating that steroids may induce hypertrophy of the fat tissue in the intracranial lipoma, causing compression of the cranial nerves passing through the cavernous sinus thereby mimicking the ocular symptoms sometimes associated with aggressive giant cell arteritis. PMID- 10380841 TI - Reduced 25-hydroxyvitamin D levels in primary Sjogren's syndrome. Correlations to disease manifestations. AB - The purpose of this study was to explore the clinical and pathogenic significance of vitamin D metabolites in primary Sjogren's syndrome (primary SS). We measured blood concentrations of 25-hydroxyvitamin D (25 OH D) and calcitriol (1,25(OH)2D)vc in 41 patients and correlated the results with blood levels of various immune activation products, as well as with patients' clinical status. Levels of 25 OH D were slightly decreased as compared to normal controls and the reduced levels of 25 OH D were stable over the observed period of 2 years. Levels of 25 OH D correlated inversely with levels of soluble interleukin-2 receptor, status indices for global disease, total exocrine disease, surface exocrine disease, internal organ exocrine disease, and mediator-induced disease. Levels of 1,25(OH)2D varied considerably and compared to normal control values. Levels of 1,25(OH)2D did not correlate with clinical/immunopathological status. In conclusion the inverse correlations found between levels of 25 OH D and measures of clinical and immunoinflammatory status support the notion that vitamin D metabolism may be involved in the pathogenesis of primary SS. PMID- 10380843 TI - Takayasu's aortitis with dissection in systemic lupus erythematosus. AB - A forty-seven-year-old Japanese woman under treatment for systemic lupus erythematosus (SLE), complained of severe back pain. Chest X-ray and MRI showed an aneurysmal dilatation of the ascending aorta. Subsequently an aortic replacement was performed. Microscopically, the resected aorta showed Takayasu's aortitis with chronic dissection. Both aortitis and dissection are rare events in SLE patients. To our knowledge, this is the first report of Takayasu's aortitis with dissection in a patient with SLE. PMID- 10380844 TI - Pure red cell aplasia as presentation of systemic lupus erythematosus: antibodies to erythropoietin. AB - In this case report we describe two patients with pure red cell aplasia (PRCA) as an initial manifestation of systemic lupus erythematosus (SLE). Antibodies to erythropoietin were determined, by an ELISA method developed in our laboratory, in frozen serum obtained from one of the patients. A high titer of antibodies to erythropoietin was detected in serum obtained before treatment with high dose intravenous immunoglobulin (IVIG). The antibody titer declined after successful treatment. This observation suggests that antibodies to erythropoietin may contribute to the pathogenesis of SLE associated PRCA. PMID- 10380845 TI - Acute onset of dermatomyositis presenting in pregnancy with rhabdomyolysis and fetal loss. AB - We report a case of acute onset of dermatomyositis with rhabdomyolysis and myoglobinuria, which presented in the 14th week of pregnancy and resulted in spontaneous abortion of the fetus. The diagnostic work up for an underlying disease was negative and the histologic examination confirmed the diagnosis of dermatomyositis, which subsequently improved with corticosteroids. PMID- 10380846 TI - Alterations in electrophysiological activity and dye coupling of striatal spiny and aspiny neurons in dopamine-denervated rat striatum recorded in vivo. AB - We recently reported that pharmacological manipulations of the dopamine system can produce more than a 4-fold increase in dye coupling between dopaminoceptive neurons in the adult rat striatal complex. During in vivo intracellular recordings, striatal neurons in control rats and in rats that had been treated with 6-hydroxydopamine were injected with either Lucifer yellow or Neurobiotin. Only rats that exhibited severe loss (i.e., larger than approximately 95%) of striatal dopamine terminals displayed a significant increase in the incidence of dye coupling between neurons in adult striatum. Moreover, this increased coupling was present only between neurons of the same morphological cell class, i.e., among clusters of spiny neurons or between aspiny neurons. Combining intracellular labeling of spiny neurons with parvalbumin immunocytochemistry demonstrated that coupling did not occur between anatomically adjacent neurons that comprised immunocytochemically and morphologically distinct cell classes. Therefore, gap junction conductance as reflected by dye coupling appears to undergo upregulation as a consequence of compromises in nigrostriatal and mesolimbic dopamine transmission. PMID- 10380847 TI - Effects of 5,7-dihydroxytryptamine depletion of tissue serotonin levels on extracellular serotonin in the striatum assessed with in vivo microdialysis: relationship to behavior. AB - Effects of i.c.v. administration of 5,7-dihydroxytryptamine (5,7-DHT) on biochemistry and behavior were studied in awake Sprague-Dawley rats. It was found that 5,7-DHT depletion of striatal tissue levels of serotonin (5-HT) does not diminish extracellular levels until substantial depletions occur. This finding is similar to those observed after 6-hydroxydopamine lesions of the brain dopamine systems. Although varying amounts of 5,7-DHT produced serotonin depletions in striatal tissue, decreases in extracellular levels were only observed at tissue depletions greater than 60% compared to saline-injected control subjects. Thus, the effects of serotonin lesions which produce only moderate depletions may not be the result of decreased extracellular serotonin, but instead may be the result of compensatory changes in remaining neurons which maintain normal extracellular serotonin concentrations. Different degrees of striatal serotonin depletion were associated with opposite behavioral effects. Moderate levels of serotonin depletion (50-75%) produced evidence of increased anxiety, while these effects were no longer seen in rats with more severe 5-HT depletions (>75%). PMID- 10380848 TI - Action of substance P (neurokinin-1) receptor activation on rat neostriatal projection neurons. AB - Substance P (SP) acts as a neurotransmitter in the neostriatum through the axon collaterals of spiny projection neurons. However, possible direct or indirect actions of SP on the neostriatal output neurons have not been described. Targets of SP terminals within the neostriatum include interneurons, spiny neurons, afferent fibers and boutons. SP induces the release of both dopamine (DA) and acetylcholine (ACh). Since some postsynaptic actions of both DA and ACh on spiny neurons are known, we asked if activation of neostriatal NK1-class receptors is able to reproduce them. The SP NK1-receptor agonist, GR73632 (1 microM), had both excitatory and inhibitory actions on virtually all spiny neurons tested at resting potential. The excitatory action was blocked by atropine and coursed with an increase in firing rate and input resistance (R(N)). The inhibitory action was blocked by haloperidol and coursed with a reduction in firing rate and R(N). Therefore, the release of both DA and ACh induced by NK1-receptor activation modulates indirectly the excitability of the projection neurons. SP facilitates the actions of these transmitters on the spiny neuron. A residual excitatory response to the NK1-receptor agonist was observed in 30% of a sample of neurons tested in the presence of both haloperidol and atropine. The increase in R(N) that accompanied this response could be observed in the presence of 1 microM TTX or 100 microM Cd2+, suggesting a direct effect. Double labeling showed that only SP-immunoreactive neurons were facilitated by NK1-receptor activation in these conditions. PMID- 10380849 TI - Gene expression of the GAD67 and GAD65 isoforms of glutamate decarboxylase is differentially altered in subpopulations of striatal neurons in adult rats lesioned with 6-OHDA as neonates. AB - The levels of mRNAs encoding for the two isoforms of glutamate decarboxylase, GAD65 and GAD67, were measured in subpopulations of striatal neurons in adult rats depleted of dopamine as neonates with 6-OHDA and chronically injected with vehicle or with the dopamine receptor agonists apomorphine or SKF-38393. In adult rats depleted of dopamine as neonates, an increase of GAD65 and GAD67 mRNA levels was measured in the striatum. These changes were paralleled by an increase in preproenkephalin (PPE) and a decrease in preprodynorphin (PPD) mRNA levels. Quantitative analysis at the cellular level indicated that GAD67 mRNA levels were increased in PPE-labeled neurons, whereas GAD65 mRNA levels were increased in PPE unlabeled neurons. Chronic and systemic injections of apomorphine or SKF-38393 induced further increases in striatal GAD65 and GAD67 mRNA levels. These increases were only detected in the subpopulation of PPE-unlabeled neurons and were paralleled by an increase in PPD mRNA levels. The increases in GAD67, GAD65, and PPD mRNA levels induced by SKF-38393 were abolished by the administration of the D1 receptor antagonist SCH-23390. The present results provide further evidence that GAD67 and GAD65 gene expression is differentially regulated in the two subpopulations of efferent striatal neurons. They also suggest that neonatal depletions in dopamine levels induce alterations of GABA-mediated signaling in the two subpopulations of striatal efferent neurons. We speculate that these alterations are involved in the behavioral particularities exhibited by rats depleted of dopamine as neonates. PMID- 10380850 TI - Cellular responses of nucleus accumbens neurons to opiate-seeking behavior: I. Sustained responding during heroin self-administration. AB - The nucleus accumbens (NAcc) has been hypothesized to be a critical component of the circuit mediating opiate-seeking behaviors. To further explore the electrophysiological correlates of opiate-seeking behavior, we recorded neurons in the NAcc and in the medial prefrontal cortex (mPFC) of rats trained to self administer heroin for at least 2 weeks. Rats were trained to lever press (FR-1 schedule) for an intravenous (i.v.) infusion of heroin (0.06 mg/kg/injection) in an operant chamber. Spontaneous single unit activity in the NAcc and the mPFC was then recorded while animals were allowed to self-administer heroin. Our data suggest that about 20% (8/42) of the NAcc neurons studied exhibited an inhibitory response immediately after heroin self-administration. However, most of the NAcc neurons studied (76%; 32/42) were not affected during heroin self-administration. In contrast, noncontingent injection of a similar dose of heroin (0.06 mg/kg/injection) had no effect on NAcc spontaneous activity (0/6). On the other hand, passive administration of higher doses of heroin (0.2-0.6/mg/kg/injection) markedly suppressed the firing rate in 46% (6/13) of the neurons studied. These effects of heroin on NAcc activity were antagonized by systemic administration ofnaloxone (4-6 mg/kg, i.v.). Studies characterizing the responses of mPFC neurons during heroin self-administration showed that 40% (2/5) of the neurons tested exhibited an inhibitory effect immediately after heroin self administration. These data suggest that in animals well-trained to self administer heroin, only a small number (20%) of the NAcc neurons studied responded to heroin self-administration. Further research is necessary to determine whether these responses are a function of the opiate-seeking state of the animal and the mechanism(s) responsible for these effects of heroin. PMID- 10380851 TI - Facilitated NMDA receptor-mediated synaptic plasticity in the hippocampal CA1 area of dystrophin-deficient mice. AB - The contribution of the cytoskeletal membrane-associated protein dystrophin in glutamatergic transmission and related plasticity was investigated in the hippocampal CA1 area of wild-type and dystrophin-deficient (mdx) mice, using extracellular recordings in the ex vivo slice preparation. Presynaptic fiber volleys and field excitatory postsynaptic potentials (fEPSPs) mediated through N methyl-D-Aspartate receptors (NMDAr) or non-NMDAr were compared in both strains. Comparable synaptic responses were observed in wild-type and mdx mice, suggesting that basal glutamatergic transmission is not altered in the mutants. By contrast, the synaptic strengthening induced by a conditioning stimulation of either 10, 30, or 100 Hz was significantly greater in mdx mice during the first minutes posttetanus. Because the posttetanic potentiation induced in the presence of the NMDAr antagonist D-APV was not affected in the mutants, a critical role of NMDAr in this increase was suggested. The magnitude of the potentiation induced by a 30 Hz stimulation in mdx mice was normalized as compared to wild-type mice by increasing the extracellular magnesium concentration from 1.5 to 3 mM. Moreover, the transitory depression of fEPSPs induced by bath-applied NMDA (50 microM for 30s) was more sensitive to an increased extracellular magnesium concentration in wild-type than in mdx mice. Our results suggest that the absence of dystrophin may facilitate NMDAr activation in the CA1 hippocampal subfield of mdx mice, which may be partly due to a reduction of the voltage-dependent block of this receptor by magnesium. PMID- 10380852 TI - Blockade of neurokinin3 receptors antagonizes drug-induced population response and depolarization block of midbrain dopamine neurons in guinea pigs. AB - In vivo extracellular recording techniques were used to investigate the effects of neurokinin3 (NK3) receptor blockade on the pharmacological activation of midbrain dopamine (DA) neurons in the guinea pig substantia nigra (A9) and ventral tegmental area (A10). The number of spontaneously active DA cells (population response) was largely increased in A10 and A9 by acute administration of haloperidol (1 and 0.5 mg/kg i.p., respectively) and this effect was dose dependently prevented in both areas by the selective NK3 receptor antagonist SR142801 (0.3, 1, 3, and 1, 3, 10 mg/kg i.p., respectively). This compound, which was totally inactive by itself, also antagonized the increase of population response induced in A10 cells by the neurotensin receptor antagonist SR142948 (1 mg/kg i.p.) and in A9 cells by the NK2 receptor antagonist SR144190 (1 mg/kg i.p.). None of the effects of SR142801 were reproduced by SR142806, its (R) enantiomer with 240-fold lower affinity for NK3 receptors. In addition, neither SR144190 (0.3 mg/kg i.p.) nor the NK1 receptor antagonist GR205171 (1 mg/kg i.p.) affected the haloperidol-induced response. The antagonistic effects of SR142801 (3 mg/kg i.p.) were also observed on the depolarization block-related decrease of A10 cell population response evoked by repeated administration (22 days) of haloperidol. Finally, SR142801 (3 mg/kg i.p.) prevented depolarization block induced in A10 cells by acute co-administration of SR142948 and haloperidol, both on population response and on single cell firing. These results on pharmacologically induced activation and depolarization block of dopamine neurons suggest that NK3 receptors play a key role in the midbrain DA function, presumably through activation by neurokinin B. PMID- 10380853 TI - Mothers' representations of their relationships with their toddlers: links to adult attachment and observed mothering. AB - Mothers (N = 125) and their firstborn sons were studied over an 11-month period to examine relations between mothers' representations of their relationships with their children (measured at 15 months by using the Parent Development Interview [PDI]), adult representations of attachment (measured at 12 months by using the Adult Attachment Interview [AAI]), and observed mothering (measured at 15 and 21 months). Results indicate (a) that mothers classified as autonomous on the AAI scored highest on the joy-pleasure/coherence dimension of the PDI and mothers classified as dismissing on the AAI scored highest on the anger dimension of the PDI and (b) that mothers scoring higher on the joy-pleasure/coherence dimension of the PDI engaged in less negative and more positive mothering. PMID- 10380854 TI - Visual control of reaching and grasping in infants. AB - The role of visual input during reaching and grasping was evaluated. Groups of infants (5, 7, and 9 months old) and adults reached for an illuminated object that sometimes darkened during the reach. Behavioral and kinematic measures were assessed during transport and grasp. Both infants and adults could complete a reach and grasp to a darkened object. However, vision was used during the reach when the object remained visible. Infants contacted the object more often when it remained visible, though they had longer durations and more movement units. In contrast, adults reached faster and more precisely during transport and grasp when the object remained visible. Thus, continuous sight of the object was not necessary, but when it was available, infants used it for contacting the object whereas adults used it to reach and grasp more efficiently. PMID- 10380855 TI - Predicting boys' social acceptance and aggression: the role of mother-child interactions and boys' beliefs about peers. AB - Seven- to 9-year-old boys (N = 177) and their mothers participated in this study in which the associations between boys' experiences with their mothers, their beliefs about familiar and unfamiliar peers, and their peer adjustment were examined across a 2-year period. Boys' negative behavior with mothers was associated with their having more negative beliefs about familiar and unfamiliar peers and with their being more aggressive and less well-liked. Beliefs about familiar peers predicted changes in boys' social acceptance, whereas negative beliefs about unfamiliar peers predicted changes in aggression. In addition, boys' beliefs about peers changed in response to their social experience. The implications of these findings for children's social development are discussed. PMID- 10380856 TI - Feeding-based arousal effects on visual recognition memory in early infancy. AB - Arousal effects on a 1-trial visual recognition paired-comparison task were studied at newborn, 1-month, and 4-month test ages. Infants were tested before and after feeding, with arousal assumed to be lower after feeding. Newborns and 1 month-olds shifted from a familiarity preference before feeding to a novelty preference after feeding. A control group tested only after feeding confirmed that this shift was not due to increased stimulus exposure from the prefeeding test. By 4 months, infants showed novelty preferences independent of feeding. This age by arousal interaction for recognition memory extends previous knowledge by including endogenous arousal with age, stimulus, and length of exposure as contributors to familiarity-novelty preferences. It also extends and provides converging evidence for arousal effects on visual attention in early infancy found previously with preferential looking. A shift from subcortical to cortical dominance is supported. PMID- 10380857 TI - Development of means-end behavior in young infants: pulling a support to retrieve a distant object. AB - Three longitudinal studies are reported in which 6-8-month-old infants were tested on means-end problems involving pulling a cloth to retrieve a toy. Production of intentional means-end behavior increased between 6 and 7 months, but although 6-month-olds' behavior was unaffected by the presence or absence of a toy on the cloth, 7-month-olds more often produced means-end sequences when a toy could be retrieved. Infants' performance remained the same when the cloth was either attached to the toy or separate, suggesting that goal-subgoal conflict does not interfere with performance of means-end sequences. By 8 months, infants could appropriately adjust their means-end behavior to the distance of the toy. These results confirm J. Piaget's (1953) original description of a shift from transitional to intentional means-end behavior and suggest that development of means-end behavior involves acquisition of knowledge of appropriate means-end relations. PMID- 10380858 TI - Learning color words involves learning a system of mappings. AB - This research examines the difficulty children encounter when acquiring 2 specific sets of adjectives, color and size words, and suggests that children must acquire a system of mapping in learning these words. Children were assessed on 4 types of mappings (word-word maps, property-property maps, word-property maps, and word-word-property maps) by completing 3 color tasks. Children also participated in comparable tasks for size words. In Study 1, 13 two-year-olds were followed longitudinally at 3-week intervals. In Study 2, 56 two-year-olds participated in a cross-sectional replication. The results indicate that children acquire color maps in a characteristic order. Children demonstrated a different pattern of acquisition for size words. The results suggest that learning word associations may promote color word acquisition and that learning color words may promote selective attention to color. PMID- 10380859 TI - Relationships between parenting and adolescent adjustment over time: genetic and environmental contributions. AB - The predictive association between parenting and adolescent adjustment has been assumed to be environmental; however, genetic and environmental contributions have not been examined. This article represents one effort to examine these associations in which a genetically informative design was used. Participants were 395 families with adolescent siblings who participated in the Nonshared Environment in Adolescent Development (D. Reiss et al., 1994) project at 2 times of assessment, 3 years apart. There were 5 sibling types in 2 types of families: 63 identical twins, 75 fraternal twins, and 58 full siblings in nondivorced families and 95 full, 60 half, and 44 genetically unrelated siblings in stepfamilies. Results indicate that the cross-lagged associations between parental conflict-negativity and adolescent antisocial behavior and depressive symptoms can be explained primarily by genetic factors. These findings emphasize the need to recognize and examine the impact that adolescents have on parenting and the contribution of genetic factors to developmental change. PMID- 10380860 TI - Maternal sensitivity during infancy and subsequent life events relate to attachment representation at early adulthood. AB - A prospective longitudinal research study of 86 prematurely born children from birth to age 18 years provided empirical evidence for continuity from infancy experience to representations of attachment at age 18 years. Young adults whose representation of attachment was dismissing had been objectively observed during infancy, 16-17 years earlier, to receive less sensitive maternal care than those infants who were later judged at early adulthood to have secure or preoccupied representations. Infancy experience alone did not differentiate young adults with secure representations from those with preoccupied representations. Rather, adverse life events through age 12, particularly parental divorce, reduced the likelihood of secure representations and increased the likelihood of preoccupied representations. The absence of adverse life events did not increase the likelihood of security for those who had not experienced early sensitive caregiving. PMID- 10380861 TI - What's in a smile? AB - In positive social contexts, both adults and older infants show more Duchenne smiling (which involves high cheek raising) than non-Duchenne smiling (which does not). This study compared Duchenne and non-Duchenne smiles in early infancy for clues to their emotional significance. Infants (N = 13) from 1 to 6 months of age were videotaped weekly for 5 min in 208 face-to-face interactions with their mothers. Levels of Duchenne and non-Duchenne smiling were correlated within interactive sessions, and the 2 smiles had similar developmental trajectories. Duchenne smiles were typically preceded by non-Duchenne smiles. The results suggest these frequently contrasted types of smiles occur in similar situations and are often different temporal phases of a continuous emotional process. In contrast to adults, infant Duchenne smiles had longer durations than non-Duchenne smiles, suggesting infant smiling does not fit adult models of emotional functioning. PMID- 10380862 TI - Children's performance on pseudoword repetition depends on auditory trace quality: evidence from event-related potentials. AB - This study explored the relation between phonological short-term memory and auditory-sensory processing in 7-9-year-old children. Twenty-four participants performed a pseudoword repetition test. The mismatch-negativity (MMN) component of auditory event-related brain potentials was obtained from 9 participants with the highest and 9 participants with the lowest scores on the test. The MMN indexes short-term auditory-sensory memory, including auditory-sensory representations for speech. It was recorded to just perceptible /baga/-/baka/ bisyllabic and easily discriminable 1000/1100-Hz tone contrasts with interstimulus intervals of 350 and 2,000 ms. The high and low repeaters differed significantly in MMN amplitude to speech stimuli at the shorter interstimulus interval. Thus, the accuracy of auditory-sensory processing seems to affect phonological short-term representations in school-age children and therefore may play a role in vocabulary development. PMID- 10380863 TI - Effects of body fat on weight concerns, dating, and sexual activity: a longitudinal analysis of black and white adolescent girls. AB - Using data from a 2-year longitudinal study of 200 Black and White adolescent girls (mean age was 13.8 years at study entry), the authors investigated the implications of differences in body fat for dating and sexual activity and the implications of heterosexual activity for dieting and weight concerns. Among White girls, and Black girls with college-educated mothers, more body fat was associated with a lower probability of dating, even among nonobese girls. However, dating and sexual experience were unrelated to subsequent dieting and weight concerns. For both Blacks and Whites, body fat was the key determinant of dieting, weight dissatisfaction, and eating concerns. These findings indicate that adolescent girls' concerns about weight have a basis in real experiential differences, and efforts to promote healthy attitudes and eating habits may be more effective if the experiential implications of weight differences are taken into account. PMID- 10380864 TI - Skewed autonomy-relatedness in preadolescents' conceptions of their relationships with mother, father, and best friend. AB - Healthy adaptation within all close relationships--whether with parents, friends, or romantic partners--involves striking a balance between connectedness to and independence from the relationship partner. For some individuals, adaptation within one or more relationships is skewed, or characterized by either an excessive concern for closeness that impedes autonomy (preoccupied stance) or an excessive concern for autonomy that inhibits closeness (avoidant stance). In this study with boys and girls aged 9-14 years, children who reported a preoccupied or avoidant stance toward their mother displayed increased social impairment in the peer group over time. There were predictable associations among children's stances toward mother, father, and best friend. Children resembled their best friend in relationship stance. The study illustrates the advantages of applying common relationship constructs (e.g., autonomy-relatedness) to the study of diverse close relationships. PMID- 10380865 TI - Verbal imprecision as an index of knowledge in transition. AB - Children can be verbally imprecise when they are learning, but this phenomenon is not well documented. Verbal imprecision, anecdotally referred to as "hemming and hawing," may be indexed by restatements, comments on one's lack of knowledge, deletions of sentence constituents, and pauses. The authors examined whether they could quantify indexes of verbal imprecision and use them to predict changes in problem-solving performance. Four types of verbal imprecision were found to predict improved performance. Results were used to make inferences about processes of knowledge change. In particular, evidence suggests that adopting a new approach and rejecting an old one may be independent, and ordered, processes. Although others have drawn similar conclusions, using verbal imprecision as the data source is a relatively unique and readily accessible method for lending support to this model of knowledge change. PMID- 10380866 TI - Women and objectified body consciousness: mothers' and daughters' body experience in cultural, developmental, and familial context. AB - Hypotheses about age-related differences in objectified body consciousness (OBC; McKinley & Hyde, 1996) based on the cultural, developmental, and familial contexts of women's body experience were tested on 151 undergraduate women and their middle-aged mothers. Mothers had lower levels of surveillance (watching the body as an outside observer) and body shame (feeling one is a bad person when appearance does not meet cultural standards) than daughters. No differences were found in appearance control beliefs, body esteem, or restricted eating, even though mothers weighed more and were less satisfied with their weight than daughters. OBC was related to measures of psychological well-being in both age groups; body esteem was more strongly related to some measures of daughters' psychological well-being than mothers'. Relationships of partner and family approval and OBC and body esteem were also examined. PMID- 10380867 TI - Rule selection versus rule execution in preschoolers: an error-detection approach. AB - In 2 experiments, an error-detection approach was used to determine whether 3 year-olds' perseverative errors on the postswitch phase of the Dimensional Change Card Sort (DCCS) are due to lack of response control or representational inflexibility. In Experiment 1, 3-, 4-, and 5-year-olds watched a puppet sort perseveratively on the postswitch phase and evaluated its responses. Most 4- and 5-year-olds detected the puppet's perseverative errors, whereas most 3-year-olds failed to do so despite detecting errors on a simpler card sort. Experiment 2 revealed that 3-year-olds who failed to correctly evaluate the puppet's behavior tended to fail their own DCCS. Results imply that perseveration on the DCCS cannot be attributed to difficulty inhibiting prepotent motor responses. Instead, changes in rule use between 3 and 5 years of age are interpreted in terms of the development of representational flexibility. PMID- 10380868 TI - Children's recall 1 or 2 years after an event. AB - In Study 1, children were reinterviewed about an event they had taken part in 2 years earlier when they were 6 years old (M.-E. Pipe & J. C. Wilson, 1994). In Study 2, children were reinterviewed about an event in which they had participated 1 year earlier when they were 6 or 9 years of age (S. Gee & M.-E. Pipe, 1995). Interviews were conducted with or without cue items and distractors, as in the original studies. The amount of information reported in free recall decreased over the 1- or 2-year delays, and for 6-year-olds, there was also a small decrease in accuracy of free recall. Reinstating specific cue items in Study 2 maintained recall when attention was drawn to them, but prompting children led to a decrease in accuracy. Whereas information repeated across interviews was highly accurate, information reported for the first time at the long delays was not. PMID- 10380869 TI - Remembering children's emotions: sources of concordant and discordant accounts between parents and children. AB - Parents were asked to recall recent events that had evoked happiness, sadness, anger, and fear in their children. Children (N = 77, 2.3-6.6 years) indicated whether they remembered each event, and if so, they described the event and how it had made them feel. Agreement between parent and child concerning how the child felt varied as a function of emotion. Children agreed with their parents' emotion attributions most often for events that parents recalled as having evoked happiness and sadness, less often for fear, and least often for anger. Children disagreed with parents' attributions of happiness and sadness most often when parents and children differed concerning the attribution of children's goals. Discordant reports about children's anger were most frequent when parents and children reported conflicting goals. Discordant reports about fear were most frequent when parents and children focused on different parts of the temporal sequence surrounding the event. PMID- 10380870 TI - Internal representational models of peers: implications for the development of problematic behavior. AB - The authors investigated the relation between children's knowledge structures for peers and externalizing behavior problems. Initial levels of aggression were evaluated in 135 boys and 124 girls (Grades 1-3; 40% African American, 60% Caucasian) in Year 1 and again in Years 6 and 9. In Year 6, 3 aspects of their social knowledge structures were assessed: quality, density, and appropriateness. Results indicate that knowledge structures are related to children's concurrent levels of externalizing behaviors and that knowledge structures are related to children's concurrent levels of externalizing behaviors and predict externalizing behaviors 3 years later even after controlling for current levels of behavior. In addition, knowledge structures in Year 6 mediate the relation between aggression in Year 1 and externalizing behaviors in Year 9. The role of knowledge structures in the maintenance and growth of children's antisocial behavior is discussed. PMID- 10380871 TI - Emotional availability and attachment representations in kibbutz infants and their mothers. AB - Three components of the attachment transmission model were examined in 48 kibbutz dyads from 2 kibbutz sleeping arrangements: communal and home-based. Concurrent assessments used the Strange Situation procedure (M. D. Ainsworth, M. C. Blehar, E. Waters, & S. Wall, 1978) for infants' attachment relations, the Adult Attachment Interview (C. George, N. Kaplan, & M. Main, 1985) for mothers' attachment representations, and the Emotional Availability Scales (Z. Biringen, J. L. Robinson, & R. N. Emde, 1993) for emotional availability in the dyads. Security of infants' attachment relations as well as autonomy of mothers' attachment representations were associated with higher emotional availability scores. In addition, significantly poorer emotional availability was found in dyads in which infants were insecurely attached and mothers were nonautonomous. Results also indicate that in the ecology of collective sleeping, the associations between the experience of emotional availability in the dyads and infants' and mothers' attachment may have been disrupted. PMID- 10380873 TI - Person versus process praise and criticism: implications for contingent self worth and coping. AB - Conventional wisdom suggests that praising a child as a whole or praising his or her traits is beneficial. Two studies tested the hypothesis that both criticism and praise that conveyed person or trait judgments could send a message of contingent worth and undermine subsequent coping. In Study 1, 67 children (ages 5 6 years) role-played tasks involving a setback and received 1 of 3 forms of criticism after each task: person, outcome, or process criticism. In Study 2, 64 children role-played successful tasks and received either person, outcome, or process praise. In both studies, self-assessments, affect, and persistence were measured on a subsequent task involving a setback. Results indicated that children displayed significantly more "helpless" responses (including self-blame) on all dependent measures after person criticism or praise than after process criticism or praise. Thus person feedback, even when positive, can create vulnerability and a sense of contingent self-worth. PMID- 10380872 TI - Early metalinguistic competence: speech monitoring and repair behavior. AB - This article examines 2-3-year-olds' responses to specific (e.g., "Where did he go?") and neutral (e.g., "What?") requests for clarification. The focus is on children's ability to locate the linguistic errors that provoked neutral questions of clarification and their success in providing appropriate repair. It is argued that this behavior provides evidence for a speech monitor that detects errors and enables repair. Contrary to A. Karmiloff-Smith's (1992) claim, control over the production and comprehension of specific structures does not necessarily precede monitoring. Rather, metalinguistic abilities that are implicated in speech monitoring develop simultaneously with the acquisition of primary linguistic behavior and do not require awareness. It is claimed that such metaprocesses are fundamental to the use of language as a vehicle for the expression of intentional content. PMID- 10380874 TI - Infant preferences for attractive faces: a cognitive explanation. AB - Research on infant face perception has shown that infants' preferences for attractive faces exist well before socialization from parents, peers, and the media can affect these preferences. Four studies assessed a cognitive explanation for the development of attractiveness preferences: cognitive averaging and infant preferences for mathematically averaged faces, or prototypes. Studies 1 and 2 demonstrated that both adults and 6-month-old infants prefer prototypical, mathematically averaged faces. Studies 3 and 4 demonstrated that 6-month-olds can abstract the central tendency from a group of naturalistic faces. Taken together, the studies suggest that infants' preferences for attractive faces can be explained by general information-processing mechanisms. PMID- 10380875 TI - Infant attention and the development of smooth pursuit tracking. AB - The effect of attention on smooth pursuit and saccadic tracking was studied in infants at 8, 14, 20, and 26 weeks of age. A small rectangle was presented moving in a sinusoidal pattern in either the horizontal or vertical direction. Attention level was distinguished with a recording of heart rate. There was an increase across age in overall tracking, the gain of the smooth pursuit eye movements, and an increase in the amplitude of compensatory saccades at faster tracking speeds. One age change was an increase in the preservation of smooth pursuit tracking ability as stimulus speed increased. A second change was the increasing tendency during attentive tracking to shift from smooth pursuit to saccadic tracking when the stimulus speed increased to the highest velocities. This study shows that the development of smooth pursuit and targeted saccadic eye movements is closely related to the development of sustained attention in this age range. PMID- 10380876 TI - After-school activities and the development of low-income urban children: a longitudinal study. AB - After-school activities of 194 African American and White children from low income households were studied from 3rd to 5th grade to determine relations with (a) child, family, and contextual variables and (b) children's adjustment over time. Girls were more likely to engage in academic activities and socializing, whereas boys were more likely to play coached sports. Children who attended after school programs spent more time on academic and extracurricular activities, whereas children in informal care settings spent more time watching TV and hanging out. Evidence of transactional relations between after-school activities and child adjustment was found. Time spent in activities between 3rd and 5th grades was related to children's adjustment in 5th grade. In addition, child adjustment measured in 3rd grade was associated with time in different activities in 5th grade. PMID- 10380877 TI - Mutant p53 proteins stimulate spontaneous and radiation-induced intrachromosomal homologous recombination independently of the alteration of the transactivation activity and of the G1 checkpoint. AB - We report here a systematic analysis of the effects of different p53 mutations on both spontaneous and radiation-stimulated homologous recombination in mouse L cells. In order to monitor different recombination pathways, we used both direct and inverted repeat recombination substrates. In each line bearing one of these substrates, we expressed p53 proteins mutated at positions: 175, 248 or 273. p53 mutations leading to an increased spontaneous recombination rate also stimulate radiation-induced recombination. The effect on recombination may be partially related to the conformation of the p53 protein. Moreover, p53 mutations act on recombination between direct repeats as well as between inverted repeats indicating that strand invasion mechanisms are stimulated. Although all of the p53 mutations affect the p53 transactivation activity measured on the WAF1 and MDM2 gene promoters, no correlation between the transactivation activity and the extent of homologous recombination can be drawn. Finally, some p53 mutations do not affect the G1 arrest after radiation but stimulate radiation-induced recombination. These results show that the role of p53 on transactivation and G1 cell cycle checkpoint is separable from its involvement in homologous recombination. A direct participation of p53 in the recombination mechanism itself is discussed. PMID- 10380878 TI - Bin1 functionally interacts with Myc and inhibits cell proliferation via multiple mechanisms. AB - The tumor suppressor Bin1 was identified through its interaction with the N terminal region of Myc which harbors its transcriptional activation domain. Here we show that Bin1 and Myc physically and functionally associate in cells and that Bin1 inhibits cell proliferation through both Myc-dependent and Myc-independent mechanisms. Bin1 specifically inhibited transactivation by Myc as assayed from artificial promoters or from the Myc target genes ornithine decarboxylase (ODC) and alpha prothymosin (pT). Inhibition of ODC but not pT required the presence of the Myc binding domain (MBD) of Bin1 suggesting two mechanisms of action. Consistent with this possibility, a non-MBD region of Bin1 was sufficient to recruit a repression function to DNA that was unrelated to histone deacetylase. Regions outside the MBD required for growth inhibition were mapped in Ras cotransformation or HepG2 hepatoma cell growth assays. Bin1 required the N terminal BAR domain to suppress focus formation by Myc whereas the C-terminal U1 and SH3 domains were required to inhibit adenovirus E1A or mutant p53, respectively. All three domains contributed to Bin1 suppression of tumor cell growth but BAR-C was most crucial. These findings supported functional interaction between Myc and Bin1 in cells and indicated that Bin1 could inhibit malignant cell growth through multiple mechanisms. PMID- 10380879 TI - Stage-specific changes in SR splicing factors and alternative splicing in mammary tumorigenesis. AB - Using a mouse model of mammary gland development and tumorigenesis we examined changes in both alternative splicing and splicing factors in multiple stages of mammary cancer. The emphasis was on the SR family of splicing factors known to influence alternative splicing in a wide variety of genes, and on alternative splicing of the pre-mRNA encoding CD44, for which alternative splicing has been implicated as important in a number of human cancers, including breast cancer. We observed step-wise increases in expression of individual SR proteins and alternative splicing of CD44 mRNA during mammary gland tumorigenesis. Individual preneoplasias differed as to their expression patterns for SR proteins, often expressing only a sub-set of the family. In contrast, tumors demonstrated a complex pattern of SR expression. Little difference was observed between neoplasias and their metastases. Alternative splicing of CD44 also changed through the disease paradigm such that tumors produced RNA containing a mixture of variable exons, whereas preneoplasias exhibited a more restricted exon inclusion pattern. In contrast, other standard splicing factors changed little in either concentration or splicing pattern in the same cells. These data suggest alterations in relative concentrations of specific splicing factors during early preneoplasia that become more pronounced during tumor formation. Given the ability of SR proteins to affect alternative processing decisions, our results suggest that a number of pre-mRNAs may undergo changes in alternative splicing during the early and intermediate stages of mammary cancer. PMID- 10380880 TI - STAT activation by the PDGF receptor requires juxtamembrane phosphorylation sites but not Src tyrosine kinase activation. AB - Activation of the platelet-derived growth factor (PDGF) receptor tyrosine kinase induces tyrosine phosphorylation of Signal Transducer and Activator of Transcription (STAT) proteins. Since the PDGF receptor also activates the Src tyrosine kinase, it is possible that Src mediates tyrosine phosphorylation of STATs in PDGF-treated cells. Consistent with a role for Src in STAT activation, we found that a PDGF receptor juxtamembrane tyrosine residue required for Src activation is necessary and sufficient for activation of STATs 1 and 3. To test the Src requirement further, we made other mutations in the PDGF receptor juxtamembrane region that increased or decreased Src binding. In epithelial and fibroblast cells, PDGF activated STAT1, 3 and 6 in the absence of detectable binding and activation of Src. In addition, PDGF induced c-myc RNA expression and DNA synthesis even though Src was not detectably activated. The activation of MAP kinase and the induction of c-fos gene expression both correlated with STAT but not Src activation by the receptor. We conclude that juxtamembrane tyrosine phosphorylation is necessary for both Src tyrosine kinase and STAT activation by the betaPDGF receptor, but that both processes are regulated independently by this region. PMID- 10380881 TI - Conditional expression of the ErbB2 oncogene elicits reversible hyperplasia in stratified epithelia and up-regulation of TGFalpha expression in transgenic mice. AB - The ErbB2 receptor tyrosine kinase (RTK) is expressed in basal cells of squamous epithelia and the outer root sheath of hair follicles. We previously showed that constitutive expression of activated ErbB2 directed to these sites in the skin by the keratin 14 (K14) promoter produces prominent hair follicle abnormalities and striking skin hyperplasia in transgenic mice. However, perinatal lethality precluded the establishment of a transgenic line for analysis of ErbB2 function in adult animals. To investigate the significance of ErbB2 signaling in epithelial tissues during and post development, we developed a K14-rtTA/TetRE ErbB2 'Tet-On' bitransgenic mouse system. These mice were normal until the ErbB2 transgene was induced by exposure to doxycycline (Dox). Prenatal induction resulted in perinatal death. Postnatally, ErbB2 transgene expression was observed at 4 h after the initiation of Dox, and reached a plateau at 24 h. Skin hyperplasia followed after 2 days and these changes reverted to normal upon Dox withdrawal. In adults, as in the neonates, prolonged ErbB2 induction caused prominent skin and hair follicle hyperplasias. Severe hyperplasias in the cornea, eye lids, tongue and esophagus were also observed. ErbB2 transgene induction was accompanied by increased expression of TGFalpha, a ligand of epidermal growth factor receptor (EGFR), and to a lesser extent, EGFR, further enhancing RTK signal transduction. We conclude that ErbB2 plays important roles in both development and maintenance of hair follicles and diverse squamous epithelia and that this ligand-inducible and tissue-specific 'Tet-On' transgenic mouse system provides a means to study transgenes with perinatal toxicity. PMID- 10380882 TI - MBP1: a novel mutant p53-specific protein partner with oncogenic properties. AB - Using a yeast two-hybrid screening strategy with a common tumour-derived p53 mutant as bait, we identified several mutant p53-interacting partners including the known proteins wild-type (wt) p53, hUBC9 and GBP/PIAS1. In addition, a novel protein partner was identified which we have termed MBP1, for Mutant p53-Binding Protein 1. MBP1 is a new member of the emerging fibulin gene family, which currently comprises fibulin-1, fibulin-2 and S1-5. Expression of MBP1 mRNA is differentially regulated both temporally during development of the mouse embryo and in a tissue-specific manner within the adult. Specific interaction between MBP1 and mutant p53 was illustrated by both two-hybrid analysis in yeast and co immunoprecipitation in mammalian cells. MBP1 displayed the following order of binding specificity towards different p53 forms: H175 > G281 > H273 > or = W248>wt p53. Thus, MBP1 appears to bind preferentially to p53 mutants of the 'structural' rather than 'contact' class, reflecting a potential bias towards those mutants having a significant alteration in conformation from that assumed by wt p53. We propose that MBP1 is the product of a candidate oncogene as rates of both neoplastic transformation and tumour cell growth were shown to be significantly enhanced when the protein is ectopically overexpressed. Furthermore, MBP1 may play a role in determining if a 'gain of function' effect is seen with certain p53 mutants. PMID- 10380883 TI - Effect of transition metals on binding of p53 protein to supercoiled DNA and to consensus sequence in DNA fragments. AB - Recently we have shown that wild-type human p53 protein binds preferentially to supercoiled (sc) DNA in vitro in both the presence and absence of the p53 consensus sequence (p53CON). This binding produces a ladder of retarded bands on an agarose gel. Using immunoblotting with the antibody DO-1, we show that the bands obtained correspond to ethidium-stained DNA, suggesting that each band of the ladder contains a DNA-p53 complex. The intensity and the number of these hands are decreased by physiological concentrations of zinc ions. At higher zinc concentrations, binding of p53 to scDNA is completely inhibited. The binding of additional zinc ions to p53 appears much weaker than the binding of the intrinsic zinc ion in the DNA binding site of the core domain. In contrast to previously published data suggesting that 100 microM zinc ions do not influence p53 binding to p53CON in a DNA oligonucleotide, we show that 5-20 microM zinc efficiently inhibits binding of p53 to p53CON in DNA fragments. We also show that relatively low concentrations of dithiothreitol but not of 2-mercaptoethanol decrease the concentration of free zinc ions, thereby preventing their inhibitory effect on binding of p53 to DNA. Nickel and cobalt ions inhibit binding of p53 to scDNA and to its consensus sequence in linear DNA fragments less efficiently than zinc; cobalt ions are least efficient, requiring >100 microM Co2+ for full inhibition of p53 binding. Modulation of binding of p53 to DNA by physiological concentrations of zinc might represent a novel pathway that regulates p53 activity in vivo. PMID- 10380884 TI - Inhibition of p38 MAP kinase increases okadaic acid mediated AP-1 expression and DNA binding but has no effect on TRE dependent transcription. AB - By performing in vitro kinase assays we found in papilloma producing 308 mouse keratinocytes that okadaic acid elevated activities of extracellular signal regulated kinase (ERK) 1/2, c-Jun N-terminal kinases (JNKs) and p38 mitogen activated protein kinases (MAPKs). This okadaic acid mediated activation of MAP kinases correlated with increased AP-1 binding to a consensus TPA responsive element (TRE) and elevated TRE dependent transcription. To determine the role of p38 MAP kinases in these processes we employed the specific p38 MAP kinase inhibitor SB 203580. Using orthophosphate labeling we showed a decrease in phosphorylation of MAPK activated protein kinase-2 (MAPKAP-K2) indicating reduced activity of p38 MAPKs utilizing this kinase as substrate. In contrast, we found that SB 203580 raised activities of ERK-1/2 and JNKs. Electrophoretic mobility shift assays revealed an increase in TRE binding activity in response to SB 203580 most likely resulting from increased expression of the major TRE binding components JunD and FosB as indicated by Western blot analyses. Increased TRE DNA binding failed to lead to increased transactivation correlating with the inability of SB 203580 to increase phosphorylation of these AP-1 proteins. These data indicate that SB 203580 sensitive p38 MAP kinases are not involved in okadaic acid mediated increases in TRE DNA binding and transactivation. PMID- 10380885 TI - p53 and Egr-1 additively suppress transformed growth in HT1080 cells but Egr-1 counteracts p53-dependent apoptosis. AB - The human fibrosarcoma cell line, HT1080, clone H4, was used to determine if the transformation suppressive functions of p53 and Egr-1 have the same underlying mechanism. This cell line expresses only mutant p53 and no detectable Egr-1. H4 clones stably expressing Egr-1 are less transformed in proportion to the level of Egr-1 expressed, acting through the induction of the TGFbeta1 gene. Here, H4 cells and the highest Egr-1 expressing clone were transfected with a vector expressing normal human p53 to derive stable clones expressing p53. The expression of p53 in H4 cells inhibited transformed growth and reduced tumorigenicity. The effect of coexpression of both p53 and Egr-1 was additive, producing cell lines with 30% of normal growth rate and sevenfold reduced tumorigenicity compared with control lines. These results indicated that each factor may act independently by different pathways, although each additively increased the level of p21WAF1 cell cycle inhibitor. However, exposure of the H4 derived cells to UV-C irradiation produced contrasting effects. Cell cycle analyses showed that the presence of p53 was associated with loss of the G1 and S cells to apoptosis after irradiation. In contrast, the expression of Egr-1 increased entry into S/G2 phase of the cell cycle with little apoptosis via a mechanism involving elevated FAK and low caspase activities. Apoptosis was observed only in the cell lines that expressed no Egr-1, especially those expressing wt-p53, and was preceded by high caspase activity. In summary, Egr-1 suppressed transformation and counteracted apoptosis by the coordinated activation of TGFbeta1, FN, p21 and FAK, leading to enhanced cell attachment and reduced caspase activity. In the doubly expressing cell line, the survival effect of Egr-1 was dominant over the apoptotic effect of p53. PMID- 10380886 TI - Cooperation of Myb and Myc proteins in T cell lymphomagenesis. AB - The v-Myb oncogene causes late onset T cell lymphomas when expressed in the T cell lineage of transgenic mice. In order to define the cellular mutations cooperating with s-Myb to cause lymphomas, we have infected v-Myb transgenic mice with Moloney murine leukemia virus (M-MuLV). Tumor formation is significantly accelerated from a mean age of onset of 60 weeks in uninfected vMyb transgenics to 13 weeks in infected vMyb transgenics. We studied the loci into which the M MuLV had inserted, and found that in 73% of animals, either the c-myc or the N myc genes had been disrupted and deregulated. Therefore, v-myb and c-myb can cooperate to induce T cell lymphomas. PMID- 10380887 TI - Oncogenic potential of Hsp72. AB - Hsp72 is the major heat shock-inducible protein capable of protecting cells from a variety of stresses. In non-transformed cells at normal conditions Hsp72 is expressed at very low levels. It is, however, present at elevated levels in the major fraction of tumors and in many transformed cell lines. It is commonly assumed that in tumor cells the expression of Hsp72 at elevated levels is the consequence of oncogenic transformation. In the present study we addressed an alternative possibility that Hsp72 plays an active role in the process of oncogenic transformation. We report here that when Hsp72 was expressed in the Rat 1 fibroblasts either constitutively or from an adenovirus-based construct, cells become oncogenically transformed by the following criteria: loss of contact inhibition and formation of foci characteristic for oncogenically transformed cells; acquisition of the ability to grow in an anchorage-independent manner and to form colonies in soft agar; generation of tumors upon injection into mice. Furthermore, we also report that turning off the Hsp72 expression led to the reversal of the transformed phenotype. We also show that oncogenic potential of Hsp72 is confined in its peptide binding domain since the expression of this domain alone was sufficient for oncogenic transformation of Rat-1 cells. PMID- 10380888 TI - Effect of p21waf1/cip1 transgene on radiation induced apoptosis in T cells. AB - The cyclin kinase inhibitor p21WAF1/Cip1 is upregulated by the tumor suppressor p53. While p21 is central for the G-1 arrest mediated by p53, it is still unclear if p21 also functions as a downstream effector of p53 dependent apoptosis. Apoptosis induced by DNA damage but not dexamethasone is p53 dependent in thymocytes. To investigate the physiological role of p21 in apoptosis, we have generated transgenic mice in which the p21 transgene is targeted for restricted expression in the T cell lineage. Thymocytes from p21 transgenic mice were hypersensitive to cell death induced by DNA damaging agents such as ionizing radiation and UV, but not be dexamethasone. Irradiated p21 transgenic thymocytes had approximately twofold more apoptotic cells as compared to irradiated age matched littermate control mice. Radiation induced death is comparable in thymocytes from p21 + Bcl2 + double transgenic mice and age matched littermate controls, indicating that the Bcl2 transgene rescues the radiation hypersensitivity imposed by p21. However, thymocytes from p53-/- mice even when they expressed the p21 transgene, were resistant to death induced by radiation. Together these results show that thymocytes from p21 transgenic mice are hypersensitive to radiation induced programmed cell death and suggest that the radiation hypersensitivity of p21 transgenic thymocytes involves p53 dependent pathway and signals in addition to p21. PMID- 10380889 TI - Early cellular abnormalities induced by RET/PTC1 oncogene in thyroid-targeted transgenic mice. AB - The RET/PTC1 oncogene, a rearranged form of the RET proto-oncogene, has been reported to be associated with human papillary thyroid carcinomas. We have shown that targeted expression of RET/PTC1 in the thyroid gland leads to the development of thyroid carcinomas in transgenic mice with histologic and cytologic similarities to human papillary thyroid carcinoma. To further investigate how RET/PTC1 expression contributes to the pathogenesis of papillary thyroid tumor, the time of tumor onset and the early phenotypic consequences of RET/PTC1 expression in thyrocytes were determined. All high copy transgenic mice developed bilateral thyroid tumors as early as 4 days of age. At embryological days 16-18, increased proliferation rate, distorted thyroid follicle formation and reduced radioiodide concentrating activity were identified in transgenic embryos. The reduced radioiodide concentrating activity was attributed to decreased expression of the sodium-iodide symporter. Our study showed that RET/PTC1 not only increased proliferation of thyrocytes, it also altered morphogenesis and differentiation. These findings provide a model for the role of RET/PTC1 in the formation of abnormal follicles with reduced iodide uptake ability observed in human papillary thyroid carcinoma. PMID- 10380890 TI - Fluorescent cDNA microarray hybridization reveals complexity and heterogeneity of cellular genotoxic stress responses. AB - The fate of cells exposed to ionizing radiation (IR) may depend greatly on changes in gene expression, so that an improved view of gene induction profiles is important for understanding mechanisms of checkpoint control, repair and cell death following such exposures. We have used a quantitative fluorescent cDNA microarray hybridization approach to identify genes regulated in response to 7 irradiation in the p53 wild-type ML-1 human myeloid cell line. Hybridization of the array to fluorescently-labeled RNA from treated and untreated cells was followed by computer analysis to derive relative changes in expression levels of the genes present in the array, which agreed well with actual quantitative changes in expression. Forty-eight sequences, 30 not previously identified as IR responsive, were significantly regulated by IR. Induction by IR and other stresses of a subset of these genes, including the previously characterized CIP1/ WAF1, MDM2 and BAX genes, as well as nine genes not previously reported to be IR responsive, was examined in a panel of 12 human cell lines. Responses varied widely in cell lines with different tissues of origin and different genetic backgrounds, highlighting the importance of cellular context to genotoxic stress responses. Two of the newly identified IR-responsive genes, FRA-1 and ATF3, showed a p53-associated component to their IR-induction, and this was confirmed both in isogenic human cell lines and in mouse thymus. The majority of the IR responsive genes, however, showed no indication of p53-dependent regulation, representing a potentially important class of stress-responsive genes in leukemic cells. PMID- 10380891 TI - TRANCE is a TNF family member that regulates dendritic cell and osteoclast function. AB - Tumor necrosis factor (TNF)-related activation-induced cytokine (TRANCE) is a new member of the TNF family emerging as a key regulator of the immune system and of bone development and homeostasis. TRANCE is expressed on activated T cells and activates mature dendritic cells (DC), suggesting that it plays a role in the T cell-DC interaction during an immune response. Furthermore, TRANCE is expressed on osteoblasts stimulated with vitamin D3, dexamethasone, and parathyroid hormone. TRANCE, when expressed on osteoblasts, induces osteoclastogenesis and osteoclast activation, suggesting that it links known calciotropic hormones to bone resorption. TRANCE mediates its effects via the TRANCE-receptor (TRANCE R/RANK), whereas its activity can be inhibited by the soluble decoy receptor osteoprotegerin/osteoclast inhibitory factor (OPG/OCIF). OPG can be neutralized by another TNF-family member, the TNF-related apoptosis-inducing ligand (TRAIL), suggesting that TRANCE is part of a complex cytokine network that regulates a diverse set of functions. We will discuss the current literature describing TRANCE and its receptors and its role in controlling DC and osteoclast function. PMID- 10380892 TI - Integrin-mediated signaling in human neutrophil functioning. AB - Integrins are important signal transducers for virtually all neutrophil functions. Although a variety of signals ultimately result in integrin activation, the intracellular targets of integrin-initiated signals are poorly delineated to date. Polymorphonuclear (PMN) leukocyte responses to inflammation are dependent on both the stimulants and the extracellular environment encountered. Integrin ligation, by cell-cell or cell-matrix interactions, activates a variety of signaling cascades. These events dictate the nature of PMN responses to the encountered stimulus. The complex system of effector molecule recruitment and permissive signaling by integrins serves to strictly regulate PMN functions such as cell adhesion, motility, oxidant production, and protein synthesis. Moreover, there is evidence that cross-talk between integrins exists to prime integrin populations for subsequent functioning. This review summarizes the current understanding of signaling mechanisms for integrin priming and activation. In this connection, the role of specific signaling molecules in key PMN functions are examined. PMID- 10380893 TI - Monocyte CD14: a multifunctional receptor engaged in apoptosis from both sides. AB - Like all other immune system cells, monocytes and macrophages may undergo apoptotic cell death in response to specific triggers and mediators or as a consequence of aging. However, factors inducing apoptosis and the involved cellular and molecular mechanisms are much better investigated and understood for lymphocytes. Th2 cell-derived cytokines such as interleukin-4 (IL-4) are able to induce monocyte apoptosis most effectively. This process is preceded by down regulation of the CD14 surface receptor. Mediators such as lipopolysaccharide (LPS) suppress and postpone apoptosis in parallel with up-regulation of CD14. Macrophages are rather resistant against apoptotic damage, and factors able to evoke apoptosis in monocytes are often ineffective in macrophages. Resistance of macrophages against apoptotic triggers may be beneficial for inflammatory processes where macrophages are engaged and needed as phagocytes for ingestion and removal of moribund cells. The multifunctional CD14 receptor of monocytes/macrophages is supposed to be involved in the apoptotic network on both sides: as a surface molecule of monocytes that can promote survival and antagonize apoptosis and as a recognition receptor of macrophages that enables or supports interaction with apoptotic cells. PMID- 10380894 TI - Differential production of MCP-1 and cytokine-induced neutrophil chemoattractant in the ischemic brain after transient focal ischemia in rats. AB - Chemokines have been shown to play an important role in leukocyte infiltration into ischemic lesions. Recently, the increased expression of monocyte chemoattractant protein-1 (MCP-1) and cytokine-induced neutrophil chemoattractant (CINC) was observed in experimental stroke models where infiltrated leukocytes were supposed to induce tissue injury, however, the protein level and time course of these chemokines have not been fully elucidated. Therefore, we analyzed the time-dependent production of MCP-1 and CINC in the rat brain after transient middle cerebral artery occlusion (MCAO) by means of specific enzyme-linked immunosorbent assay systems. The MCP-1 levels in the ipsilateral hemispheres increased from 6 h, peaked at 2 days, and thereafter gradually decreased. The peak MCP-1 concentration was 89.2+/-28.2 ng/g tissue wet weight (mean +/- SEM, n = 5, 49.3-fold greater than the contralateral value at the same time, P < 0.05), which is supposed to be high enough to exert its biological effects. In contrast, the maximum CINC concentration that corresponded to 2.9+/-0.7 ng/g tissue wet weight (mean +/- SEM, n = 5, 55.0-fold greater than the contralateral value at the same time, P < 0.05), was observed at 6 h. In addition, we confirmed the temporal profile of leukocyte subtypes that infiltrated into the ischemic brain, thus, neutrophil infiltration occurred at early stages (1-3 days), followed by massive infiltration of macrophages at later stages (2-7 days). These studies suggest that MCP-1 in cerebral ischemia actually plays a significant role in the migration of macrophages into the lesion and that the differential temporal production of these chemokines contributes to the regulation of infiltrated leukocyte subtypes. PMID- 10380895 TI - Enhancement of leukocyte response to lipopolysaccharide by secretory group IIA phospholipase A2. AB - Secretory nonpancreatic group IIA phospholipase A2 (sPLA2), a lipolytic enzyme found in plasma, is thought to play an important role in inflammation. In patients with sepsis, a strong positive correlation is observed between the plasma level of sPLA2 and poor clinical outcome in sepsis. We have thus asked whether sPLA2 could play a role in enabling responses of cells to bacterial lipopolysaccharide (LPS), a key contributor to sepsis. In the presence of sPLA2, cellular responses to LPS were significantly increased. This was demonstrated in assays of LPS-stimulated interleukin-6 (IL-6) production in whole blood and binding of freshly isolated human polymorphonuclear neutrophils (PMN) to fibrinogen-coated surfaces. We further found that sPLA2 enhanced binding of labeled LPS to PMN, and that the sPLA2-mediated cell responses to LPS were all blocked by monoclonal antibodies directed against membrane CD14. Two properties ofsPLA2 may contribute to its activity to mediate responses to LPS. sPLA2 appears to bind LPS because pre-exposure of sPLA2 to LPS led to a dose-dependent increase in its ability to hydrolyze phospholid substrate, and incubation of sPLA2 with BODIPY-LPS micelles resulted in enhanced fluorescence, presumably from the disaggregation of the LPS aggregates. Additional studies demonstrated that the esterolytic function of sPLA2 is also needed both for the disaggregation of LPS and CD14-dependent cell stimulation. The precise mechanisms by which LPS-binding and esterolytic activity contribute to sPLA2 activity are not clear but our data strongly suggest that these activities result in interaction of LPS with CD14 and subsequent cell activation. PMID- 10380896 TI - Vitamin E protects against polymorphonuclear leukocyte-dependent adhesion to endothelial cells. AB - We evaluated the effects of alpha-Toc on surface expression of CD11b/CD18 on polymorphonuclear leukocytes (PMN) stimulated with N-formyl-methionyl-leucyl phenylalanine (fMLP) and oxidized low-density lipoprotein (oxLDL). Incubation of PMN with fMLP (1 microM) or oxLDL (100 microg/mL) increased CD11b/CD18 expression; pretreatment with alpha-Toc reduced in a dose-dependent manner. PMN obtained from healthy adults ingesting 600 mg alpha-Toc per day for 10 days were similarly incubated with fMLP or oxLDL; the surface level of CD11b/CD18 was inversely correlated with serum alpha-Toc concentrations. Adherence of PMN to human umbilical vein endothelial cells was increased by fMLP or oxLDL stimulation but reduced by alpha-Toc pretreatment or anti-CD18 monoclonal antibodies. cAMP dependent protein kinase (PKA) and protein kinase C (PKC) activity in PMN was also assayed. A PKC inhibitor, but not a PKA inhibitor, suppressed CD11b/CD18 up regulation, and alpha-Toc slightly decreased fMLP- and oxLDL-induced activation of PKC. These results suggest that alpha-Toc may prevent inflammation by both reducing CD11b/ CD18 up-regulation and decreasing PMN-dependent adherence to EC. PMID- 10380897 TI - Cytolysis of human dendritic cells by autologous lymphokine-activated killer cells: participation of both T cells and NK cells in the killing. AB - Dendritic cells (DC) play a key role in the initiation of immune response by stimulating the naive T cells. The fate of DC after the initiation of immune response is not clearly understood. Although there are few reports implicating natural killer (NK) cells in the elimination of DC, killing of DC by LAK cells, and specifically by T cells, has not been studied. In this study, we observed that DC, generated from monocytes, in vitro in the presence of granulocyte macrophage colony-stimulating factor, interleukin-4 (IL-4), and tumor necrosis factor alpha were susceptible to cytolysis by lymphokine-activated killer (LAK) cells induced in the presence of IL-2 and IL-15 but not IL-12 alone. However, LAK cells induced by a combination of IL-12 and suboptimal dose of IL-2 were cytotoxic to DC. When purified lymphocytes were activated with IL-2, the CD8+/CD57- fraction (T-LAK), but not the CD8-/CD57+ fraction (NK-LAK) was cytotoxic to autologous DC. However, when unseparated peripheral blood mononuclear cells were used to generate LAK cells, both T-LAK and NK-LAK fractions showed equal cytotoxicity against autologous DC. Monoclonal antibodies against CD54, CD11a, and CD18 significantly inhibited the cytolysis, indicating that the killing involves the engagement of CD54 with its ligands. PMID- 10380898 TI - The synthetic non-toxic drug 2,3-dimethyl-6(2-dimethylaminoethyl)-6H-indolo-(2,3 b)quinoxaline inhibits neutrophil production of reactive oxygen species. AB - The effects of the non-toxic drug 2,3-dimethyl-6(2-dimethylaminoethyl)-6H-indolo (2,3-b)quinoxaline (B220) on the generation of reactive oxygen species froin human neutrophils were investigated. The data show that B220 inhibits neutrophil release of reactive oxygen species, as well as intracellular generation of reactive oxygen species. The inhibition is not achieved through direct oxygen radical scavenger activity of B220, and the drug has no immediate effects on the activity of the assembled oxidase. Radical production and release were inhibited by all agonists tested [i.e. the protein kinase C-activating phorbol ester phobol myristate acetate; the receptor-specific agonist N-formyl-methionyl-leucyl phenylalanine (fMLP); and serum-opsonized yeast particles] in the presence of B220. The drug also inhibits phagocytosis and fMLP-induced mobilization of granules. However, based on the fact that the effects of B220 on phagocytosis and granule mobilization are much less significant than its effect on radical production, we suggest that the signal(s) affected by B220 is (are) located mainly downstream of the point at which the signals are generated to promote oxidase activation and phagocytosis or granule secretion, respectively. PMID- 10380899 TI - Ubiquitin-proteasome system is involved in induction of LFA-1/ICAM-1-dependent adhesion of HL-60 cells. AB - Membrane-permeable proteasome inhibitors, lactacystin (LC) and N-acetyl-Leu-Leu norleucinal (ALLN), but not calpain inhibitor Z-Leu-leucinal (ZLL), prevented LFA 1/ICAM-1-dependent cellular adhesion of TPA-stimulated HL-60 cells. These proteasome inhibitors affected neither the induction of monocytic differentiation nor the accompanying protein-tyrosine phosphorylation. They suppressed the increase in the avidity of LFA-1 to ICAM-1 without changing the expression of these molecules. Immunoblotting using monoclonal antibody FK-1, which reacts specifically with polyubiquitinated proteins, demonstrated that the proteasome inhibitors caused the drastic accumulation of the polyubiquitinated proteins in the membrane fraction of TPA-treated HL-60 cells. This indicates that accompanying activation of LFA-1, TPA induces the polyubiquitination of the membrane proteins, which are rapidly degraded by proteasomes. These data taken together show that proteolysis mediated by the ubiquitin-proteasome system is a prerequisite for the induction of LFA-1-dependent adhesion of HL-60 cells. PMID- 10380900 TI - Exposure of cultured murine peritoneal macrophages to low concentrations of beryllium induces increases in intracellular calcium concentrations and stimulates DNA synthesis. AB - Exposure of humans to beryllium dusts can induce a specific form of chronic pneumonitis that consists mainly of noncaseating granulomas in the lungs. Multiple studies have documented both genetic and immune components of chronic berylliosis. Much work has focused on T cells and their reactivity in berylliosis, but less work has focused on the end effector cells in granulomatous inflammation, macrophages. Because macrophages must become activated to form granulomas, and they become activated by responding to numerous immunomodulatory signals, we investigated the effects of beryllium (BeCl2) on a central signal transduction pathway in macrophages, increases in intracellular calcium ([Ca2+]i). Exposure of cultured murine peritoneal macrophages to low, nontoxic concentrations induced successive spikes or oscillations in [Ca2+]i. Concentrations as low as 5 nM induced significant increases in [Ca2+]i. The source of the increased [Ca2+]i was exclusively extracellular in that increases in [Ca2+]i could be completely blocked by chelating extracellular Ca2+, were inhibited by the Ca2+ channel blocker verapamil, and exposure of macrophages to BeCl2 had no effect on IP3 concentrations. DNA synthesis, a Ca2+-sensitive function, was enhanced in dividing 1LN cells and induced de novo in quiescent macrophages. Furthermore, BeCl2 enhanced DNA synthesis in the absence of coexposure to the protein kinase C activator phorbol myristate acetate. These data support the hypothesis that beryllium toxicity is in part the result of altered Ca2+ metabolism in mononuclear phagocytes consequent to reversible opening of plasma membrane channels. PMID- 10380901 TI - Blockade of nitric oxide formation down-regulates cyclooxygenase-2 and decreases PGE2 biosynthesis in macrophages. AB - Elevated levels of nitric oxide (NO*) produced by expression of inducible nitric oxide synthase (iNOS/NOS type 2) and high levels of prostaglandins (PGs) generated by expression of inducible cyclooxygenase (COX-2/PGH2 synthase-2) are important mediators of immune and inflammatory responses. Previous studies have shown that endogenous levels of NO* can influence the formation of PGs. We examined the mechanism by which NO* regulates PG biosynthesis in macrophages. Treatment of a murine macrophage cell line (ANA-1) with lipopolysaccharide (LPS, 10 ng/mL) and interferon-gamma (IFN-gamma, 10 U/mL) for 20 h elicited high levels of nitrite (NO2-) and prostaglandin E2 (PGE2) that were inhibited in a dose dependent fashion by the NOS inhibitor, aminoguanidine (AG), with IC50 values of 15.06 and 0.38 microM for NO2- and PGE2, respectively. Stimulation of cultures with LPS and IFN-gamma for 20 h induced de novo iNOS protein expression that was not altered by the addition of AG (0.1, 10, or 1000 microM). In contrast, treatment of cultures with LPS and IFN-gamma for 20 h promoted COX-2 mRNA and protein expression that were decreased in a dose-dependent fashion by AG (P < 0.05 with 10 and 1000 microM). LPS and IFN-gamma-induced COX-2 protein expression was not decreased in cultures treated with AG for 2 h, illustrating that AG does not inhibit the formation of COX-2 protein. Analysis of partially purified enzyme extracts demonstrated that AG did not directly inhibit the enzymatic activity of COX. Additional experiments revealed that NO* donors (S-nitroso-N-aceytl-D-L pencillamine, SNAP, at 0.1, 10, and 1000 microM) did not induce de novo COX-2 protein expression or potentiate COX-2 expression in cells treated with LPS and/or IFN-gamma. Our results suggest that, while endogenous NO* is not required for de novo COX-2 mRNA and protein expression, NO* is necessary for maintaining prolonged COX-2 gene expression. PMID- 10380902 TI - Dysregulation of PMN antigen expression in Type 2 diabetes may reflect a generalized defect of exocytosis: influence of hypertension and microalbuminuria. AB - Defective exocytosis could underlie clinical and metabolic abnormalities in Type 2 diabetes. Because many SNARE proteins appear to be common mediators of exocytosis, we examined phorbol myristate acetate-stimulated expression of CD11b and CD69 on polymorphonuclear leukocytes (PMN) from Type 2 diabetic subjects with hypertension and microalbuminuria (D-htma), hypertension only (D-ht) or uncomplicated (D-uc), and normal controls (NC) by flow cytometry. CD11b expression was rapid (half maximal by 7 min), initially on all PMN. CD69 expression took place subsequently but on PMN that did not express CD11b. The proportion of CD11b-positive PMN at 30 min was higher in all diabetic groups than in NC. Expression of CD11b was higher and CD69 lower in D-uc and D-htma but were similar in NC and D-ht. In Type 2 diabetes the transition from the CD11b-positive to CD69-positive state is impaired. The defect in the process of CD69 expression appeared most marked in diabetic subjects with hypertension and microalbuminuria. PMID- 10380903 TI - Mycoplasma arginini enhances cytotoxicity of thioglycollate-elicited murine macrophages toward YAC-1 tumor cells through production of NO. AB - Bacterial products stimulate macrophage tumoricidal activity through release of tumor necrosis factor (TNF) and nitric oxide (NO). We show here that thioglycollate-elicited macrophages acquire cytotoxic activity when cocultured with Mycoplasma arginini-infected YAC-1 tumor cells and release TNF and NO. Fixed mycoplasma-infected cells, supernatants from infected-cell cultures, or purified heat-killed mycoplasma obtained from cell-free cultures were all able to induce TNF and NO production. Thus, the mycoplasma per se and not a product of infected cells induce the release of these molecules. Addition of prostaglandin E2 (PGE2) to the cocultures, which reduced TNF release, or antibodies to TNF, did not affect macrophage cytotoxicity nor NO release. Inhibition of NO production by L NAME or aminoguanidine reduced the cytotoxicity, and treatment with a NO donor was toxic to YAC-1 cells. These results indicate that M. arginini activates thioglycollate-elicited murine macrophages for NO and TNF release increasing their cytotoxic activity toward YAC-1 cells and that this activity is dependent on NO but not TNF release. PMID- 10380904 TI - Inhibition of microglial cell RANTES production by IL-10 and TGF-beta. AB - Using human fetal microglial cell cultures, we found that the gram-negative bacterial cell wall component lipopolysaccharide (LPS) stimulated RANTES (regulated upon activation of normal T cell expressed and secreted) production through the protein kinase C signaling pathway and that activation of transcription nuclear factor (NF)-kappaB was required for this effect. Similarly, the proinflammatory cytokines interleukin (IL)-1beta and tumor necrosis factor alpha dose-dependently stimulated microglial cell RANTES production via NF-kappaB activation. Anti-inflammatory cytokines, IL-10, and transforming growth factor (TGF)-beta sequentially inhibited LPS- and cytokine-induced microglial cell NF kappaB activation, RANTES mRNA expression, and protein release. Proinflammatory cytokines but not LPS also stimulated RANTES production by human astrocytes. These findings demonstrate that human microglia synthesize RANTES in response to proinflammatory stimuli, and that the anti-inflammatory cytokines IL-10 and TGF beta down-regulate the production of this beta-chemokine. These results may have important therapeutic implications for inflammatory diseases of the brain. PMID- 10380905 TI - Dendritic cells and MPIF-1: chemotactic activity and inhibition of endogenous chemokine production by IFN-gamma and CD40 ligation. AB - We have examined the biological activity of the CC chemokine myeloid progenitor inhibitory factor 1 (MPIF-1) on human dendritic cells. MPIF-1 has chemotactic activity on dendritic cells derived from either peripheral blood monocytes or cord blood CD34+ progenitors. However, chemokine treatment did not induce further cell activation or maturation. In addition, MPIF-1 is constitutively released by monocyte-derived dendritic cells but not macrophages or monocytes (resting or stimulated). The proinflammatory stimuli lipopolysaccharide and tumor necrosis factor alpha, which induced the release of monocyte chemotactic protein-1, macrophage inflammatory protein-1alpha (MIP-1alpha), MIP-1beta, and interleukin 8, did not affect MPIF-1 release. In contrast, CD40 ligation and interferon-gamma treatment, while stimulating the production of the other chemokines, caused a pronounced reduction of MPIF-1 transcript and protein release. Thus, in dendritic cells the regulation of the production and release of MPIF-1 is distinct in comparison to other CC and CXC chemokines. PMID- 10380906 TI - Identification of CD137 as a potent monocyte survival factor. AB - CD137 (ILA/4-1BB), a member of the tumor necrosis factor (TNF) receptor family, promotes adherence and prolongs survival of human peripheral monocytes. It induces a strong expression of macrophage colony-stimulating factor (M-CSF), an essential monocyte survival factor. Monocyte survival induced by CD137 is primarily mediated by M-CSF and to a lesser extent by granulocyte-macrophage colony-stimulating factor and IL-3. Survival and induction of M-CSF are mediated via reverse signaling through a CD137 ligand expressed constitutively by peripheral monocytes. PMID- 10380908 TI - Identification of the CD85 antigen as ILT2, an inhibitory MHC class I receptor of the immunoglobulin superfamily. AB - The CD85 molecule was originally defined at the Fifth Workshop on Leucocyte Antigens in 1993 by two monoclonal antibodies, VMP55 and GHI/75. This cell surface glycoprotein is expressed on B cells, monocytes, and subpopulations of T and natural killer (NK) cells, and particularly high levels are expressed by normal and neoplastic plasma cells and by hairy cell leukemia B cells. We affinity purified the CD85 antigen and obtained tryptic peptide sequence which indicated that this molecule might be ILT2, a recently described inhibitory major histocompatibility complex class I receptor of the immunoglobulin superfamily. This was confirmed by showing that both of the original anti-CD85 mAbs stained ILT2 transfectants. The cell signaling role demonstrated for ILT2 is consistent with the previously reported involvement of CD85 in T cell activation. PMID- 10380907 TI - Substance P primes the formation of hydrogen peroxide and nitric oxide in human neutrophils. AB - Substance P (SP), a neurotransmitter of the central and peripheral nervous system, has been implicated as a mediator of the pulmonary inflammatory response through its stimulatory effects on neutrophils. We investigated the role of SP in priming the production of reactive oxygen species by human neutrophils with the cytochrome c reduction assay and by flow cytometry using the intracellular oxidizable probe dichlorofluorescein. We also investigated SP-induced formation of nitrite and nitrate as an index of nitric oxide (NO) production. Our results indicate that SP primes two distinct pathways with respect to the induction of reactive oxygen species in the human neutrophil: the production of superoxide anion and hydrogen peroxide by the calmodulin-dependent NADPH oxidase, and the generation of NO by a constitutive NO synthase. Preincubation of neutrophils with inhibitors of calmodulin and NO synthase diminished the oxidative response in an additive fashion. These results give insight into distinct signal transduction pathways in the SP-primed neutrophil with respect to the formation of superoxide anion, hydrogen peroxide, and NO. PMID- 10380909 TI - Depletion of eosinophils in mice through the use of antibodies specific for C-C chemokine receptor 3 (CCR3). AB - We have generated rat monoclonal antibodies specific for the mouse eotaxin receptor, C-C chemokine receptor 3 (CCR3). Several anti-CCR3 mAbs proved to be useful for in vivo depletion of CCR3-expressing cells and immunofluorescent staining. In vivo CCR3 mAbs of the IgG2b isotype substantially depleted blood eosinophil levels in Nippostrongyus brasiliensis-infected mice. Repeated anti CCR3 mAb treatment in these mice significantly reduced tissue eosinophilia in the lung tissue and bronchoalveolar lavage fluid. Flow cytometry revealed that mCCR3 was expressed on eosinophils but not on stem cells, dendritic cells, or cells from the thymus, lymph node, or spleen of normal mice. Unlike human Th2 cells, mouse Th2 cells did not express detectable levels of CCR3 nor did they give a measurable response to eotaxin. None of the mAbs were antagonists or agonists of CCR3 calcium mobilization. To our knowledge, the antibodies described here are the first mAbs reported to be specific for mouse eosinophils and to be readily applicable for the detection, isolation, and in vivo depletion of eosinophils. PMID- 10380910 TI - Protein kinase C and a calcium-independent phospholipase are required for IgG mediated phagocytosis by Mono-Mac-6 cells. AB - Mono-Mac-6 (MM6) human monocytes ingest IgG-opsonized particles better than other human cell lines. We compared the phagocytic signaling pathway in MM6 with human monocytes. MM6 expressed FcgammaRI at levels similar to monocytes, whereas FcRgammaII expression was approximately double. MM6 ingested IgG-opsonized erythrocytes (EIgG) in a calcium-independent manner. Incubation of MM6 with bromoenol lactone, an inhibitor of the phagocytic phospholipase (pPL), coordinately decreased phagocytosis and pPL activity. This inhibition was overcome by exogenous arachidonic acid, suggesting that phagocytosis requires pPL activation and arachidonic acid release. MM6 phagocytosis was inhibited with staurosporine and activated with diacylglycerol, supporting a role for protein kinase C (PKC) in this process. The pPL activators mastoparan and melittin restored phagocytosis to PKC-inhibited cells, suggesting that pPL lies downstream from PKC. These results suggest that the MM6 signal transduction pathway for IgG mediated phagocytosis is similar to that of monocytes (PKC-->pPL-->arachidonic acid-->phagocytosis). The results are discussed in the context of the finding that MM6 exhibit low phagocytosis relative to monocytes and thus may represent an attractive cell line for molecular manipulation in "recovery of function" studies. PMID- 10380911 TI - Aspirin-induced increases in soluble IL-1 receptor type II concentrations in vitro and in vivo. AB - This study examined the influence of low-dose aspirin on interleukin (IL)-1alpha , IL-1 receptor antagonist (IL-1ra), and soluble receptor type II (sIL-1RII) secretion in vivo and in vitro. Blood mononuclear cells were isolated from healthy young men who ingested 81 mg of aspirin on alternate days for 2 weeks and from unmedicated controls. Aspirin had minor effects on ex vivo secretion of IL 1beta and no influence on IL-1ra. In contrast, unstimulated ex vivo secretion of sIL-1RII was over twice as high by cells from aspirin-treated subjects (1115+/ 123 vs. 460+/-77 pg/mL, P = 0.02). Lipopolysaccharide-stimulated sIL-1RII secretion was influenced similarly. Plasma sIL-1RII concentrations were 23% higher in aspirin-treated subjects (10.2+/-0.6 vs. 8.4+/-0.3 ng/mL, P = 0.03). In addition, cells from unmedicated subjects cultured in vitro with aspirin (10 microg/mL) secreted significantly greater amounts of sIL-1RII. Thus, low-dose aspirin therapy may prevent inflammation by increasing soluble receptor secretion, thereby preventing IL-1 from binding target cells. PMID- 10380912 TI - ICAM-3 (CD50) cross-linking augments signaling in CD3-activated peripheral human T lymphocytes. AB - ICAM-3 is a pan-hematopoietic, constitutive adhesion molecule. ICAM-3 binds to LFA-1 on antigen-presenting cells (APC) stabilizing the T cell-APC interaction, facilitating signaling through the CD3/TCR complex. However, recent evidence using cultured and transformed T cells suggests ICAM-3 may also function in signaling. Because ICAM-3 is constitutively expressed on resting T cells, we postulated that signaling through ICAM-3 in resting T cells represents an important costimulatory mechanism in these cells. In purified resting human T cells, cross-linking both ICAM-3 and CD3 with plate-bound antibodies resulted in a marked increase in cell size (consistent with blastogenesis), synergistically increased surface expression of CD25 and CD69, and increased T cell metabolism. Similarly, concomitant ICAM-3 and CD3 stimulation significantly (P < 0.001) increased resting human T cell phosphatidylinositol hydrolysis and phospholipase C-gamma1 phosphorylation. These results indicate that ICAM-3 augments signaling through CD3, functioning as a costimulatory molecule for resting T cells in the initial activation step. PMID- 10380913 TI - Regulation of neutrophil FcgammaRIIIb (CD16) surface expression following delayed apoptosis in response to GM-CSF and sodium butyrate. AB - When neutrophils undergo apoptosis, they lose expression of the surface receptor CD16 (FcgammaRIIIb). Thus levels of surface CD16 are good indicators of apoptotic or non-apoptotic neutrophils. Shedding of CD16 occurs via the activity of a metalloproteinase that cleaves the receptor from the plasma membrane. Granulocyte macrophage colony-stimulating factor (GM-CSF) and sodium butyrate both stimulate neutrophil gene expression, protect these cells from apoptosis, and maintain expression of surface CD16. In this report we have investigated whether these agents maintain surface expression of CD16 via (1) decreased shedding (2) increased mobilization of the internal pool of pre-formed CD16, or (3) via de novo biosynthesis of new receptor molecules. Although GM-CSF and sodium butyrate both preserved surface expression of CD16, GM-CSF actually accelerated the rate of shedding of this receptor. Maintenance of surface levels was achieved by substantial mobilization of the internal pool of CD16. Sodium butyrate, on the other hand, maintained surface expression without extensive store depletion via a mechanism that appeared to involve a decreased rate of shedding. In these experiments we could find no evidence for de novo biosynthesis of CD16 stimulated by either GM-CSF or sodium butyrate. These experiments indicate that multiple mechanisms exist for the maintenance of surface CD16 during rescue of neutrophils from apoptosis by different agents. PMID- 10380914 TI - Inhibitors of PI 3-kinase and MEK kinase differentially affect mediator secretion from immunologically activated human basophils. AB - Effects of inhibitors of PI 3-kinase and MEK kinases were investigated on histamine, leukotriene C4(LTC4), and cytokine release from human basophils stimulated with anti-IgE. The PI 3-kinase antagonists wortmannin (> 10 nM) and LY 294002 (>1 microM) strongly inhibited anti-IgE-induced release of all mediators by 40-100%. This was contrasted by the effects of the MEK kinase inhibitor PD 098059, which weakly inhibited histamine, interleukin (IL)-4, and IL-13 release but was substantially more efficacious at blocking LTC4 production (>70% at 10 microM). Previous studies have shown that arachidonic acid synthesis is controlled by MEK kinases. We observed that wortmannin, LY 294002, and PD 098059 reduce basophil ERK-1,2 activation, thus implying that, with regard to arachidonic acid metabolism, MEK kinases are a downstream target for PI-3-kinase. Our results demonstrate a universal regulatory role played by PI 3-kinases in basophil mediator production and release, whereas MEK kinase signaling is largely limited to controlling arachidonic acid metabolism. PMID- 10380916 TI - A new role for glia: generation of neurons! PMID- 10380915 TI - Tyrphostin AG-490 inhibits cytokine-mediated JAK3/STAT5a/b signal transduction and cellular proliferation of antigen-activated human T cells. AB - Janus kinase 3 (JAK3) is a cytoplasmic tyrosine kinase required for T cell development and activated by cytokines that utilize the interleukin-2 (IL-2) receptor common gamma chain (gamma(c)). Genetic inactivation of JAK3 is manifested as severe combined immunodeficiency disease (SCID) in humans and mice. These findings have suggested that JAK3 represents a pharmacological target to control certain lymphoid-derived diseases. Here we provide novel evidence that AG 490 potently inhibits the autokinase activity of JAK3 and tyrosine phosphorylation and DNA binding of signal transducer and activator of transcription 5a and 5b (STAT5a/b). Similar inhibitory effects were observed with other cytokines that use gamma(c). AG-490 also inhibited IL-2-mediated proliferative growth in human T cells with an IC50) = 25 microM that was partially recoverable. Moreover, we demonstrate that this inhibitor prevented tetanus toxoid antigen-specific T cell proliferation and expansion but failed to block activation of Zap70 or p56Lck after anti-CD3 stimulation of human T cells. Taken together, these findings suggest that AG-490 inhibits the JAK3-mediated Type II signaling pathway but not the T cell receptor-derived Type I pathway and possesses therapeutic potential for T cell-derived pathologies such as graft versus-host disease, allergy, and autoimmune disorders. PMID- 10380917 TI - Lipoprotein receptors: signaling functions in the brain? PMID- 10380918 TI - Signaling networks--do all roads lead to the same genes? PMID- 10380919 TI - Protein translocation: how Hsp70 pulls it off. PMID- 10380920 TI - Lack of BCL10 mutations in germ cell tumors and B cell lymphomas. PMID- 10380921 TI - Absence of BCL10 mutations in human malignant mesothelioma. PMID- 10380922 TI - Reeler/Disabled-like disruption of neuronal migration in knockout mice lacking the VLDL receptor and ApoE receptor 2. AB - Layering of neurons in the cerebral cortex and cerebellum requires Reelin, an extracellular matrix protein, and mammalian Disabled (mDab1), a cytosolic protein that activates tyrosine kinases. Here, we report the requirement for two other proteins, cell surface receptors termed very low density lipoprotein receptor (VLDLR) and apolipoprotein E receptor 2 (ApoER2). Both receptors can bind mDab1 on their cytoplasmic tails and are expressed in cortical and cerebellar layers adjacent to layers that express Reelin. mDab1 expression is upregulated in knockout mice that lack both VLDLR and ApoER2. Inversion of cortical layers and absence of cerebellar foliation in these animals precisely mimic the phenotype of mice lacking Reelin or mDab1. These findings suggest that VLDLR and ApoER2 participate in transmitting the extracellular Reelin signal to intracellular signaling processes initiated by mDab1. PMID- 10380923 TI - Subventricular zone astrocytes are neural stem cells in the adult mammalian brain. AB - Neural stem cells reside in the subventricular zone (SVZ) of the adult mammalian brain. This germinal region, which continually generates new neurons destined for the olfactory bulb, is composed of four cell types: migrating neuroblasts, immature precursors, astrocytes, and ependymal cells. Here we show that SVZ astrocytes, and not ependymal cells, remain labeled with proliferation markers after long survivals in adult mice. After elimination of immature precursors and neuroblasts by an antimitotic treatment, SVZ astrocytes divide to generate immature precursors and neuroblasts. Furthermore, in untreated mice, SVZ astrocytes specifically infected with a retrovirus give rise to new neurons in the olfactory bulb. Finally, we show that SVZ astrocytes give rise to cells that grow into multipotent neurospheres in vitro. We conclude that SVZ astrocytes act as neural stem cells in both the normal and regenerating brain. PMID- 10380924 TI - WRM-1 activates the LIT-1 protein kinase to transduce anterior/posterior polarity signals in C. elegans. AB - During C. elegans development, Wnt/WG signaling is required for differences in cell fate between sister cells born from anterior/posterior divisions. A beta catenin-related gene, wrm-1, and the lit-1 gene are effectors of this signaling pathway and appear to downregulate the activity of POP-1, a TCF/LEF-related protein, in posterior daughter cells. We show here that lit-1 encodes a serine/threonine protein kinase homolog related to the Drosophila tissue polarity protein Nemo. We demonstrate that the WRM-1 protein binds to LIT-1 in vivo and that WRM-1 can activate the LIT-1 protein kinase when coexpressed in vertebrate tissue culture cells. This activation leads to phosphorylation of POP-1 and to apparent changes in its subcellular localization. Our findings provide evidence for novel regulatory avenues for an evolutionarily conserved Wnt/WG signaling pathway. PMID- 10380925 TI - Diverse signaling pathways activated by growth factor receptors induce broadly overlapping, rather than independent, sets of genes. AB - We sought to explore the relationship between receptor tyrosine kinase (RTK) activated signaling pathways and the transcriptional induction of immediate early genes (IEGs). Using global expression monitoring, we identified 66 fibroblast IEGs induced by platelet-derived growth factor beta receptor (PDGFRbeta) signaling. Mutant receptors lacking binding sites for activation of the PLCgamma, PI3K, SHP2, and RasGAP pathways still retain partial ability to induce 64 of these IEGs. Removal of the Grb2-binding site further broadly reduces induction. These results suggest that the diverse pathways exert broadly overlapping effects on IEG induction. Interestingly, a mutant receptor that restores the RasGAP binding site promotes induction of an independent group of genes, normally induced by interferons. Finally, we compare the PDGFRbeta and fibroblast growth factor receptor 1; each induces essentially identical IEGs in fibroblasts. PMID- 10380926 TI - Simple, but not branched, plasmodesmata allow the nonspecific trafficking of proteins in developing tobacco leaves. AB - Leaves undergo a sink-source transition during which a physiological change occurs from carbon import to export. In sink leaves, biolistic bombardment of plasmids encoding GFP-fusion proteins demonstrated that proteins with an Mr up to 50 kDa could move freely through plasmodesmata. During the sink-source transition, the capacity to traffic proteins decreased substantially and was accompanied by a developmental switch from simple to branched forms of plasmodesmata. Inoculation of sink leaves with a movement protein-defective virus showed that virally expressed GFP, but not viral RNA, was capable of trafficking between sink cells during infection. Contrary to dogma that plasmodesmata have a size exclusion limit below 1 kDa, the data demonstrate that nonspecific "macromolecular trafficking" is a general feature of simple plasmodesmata in sink leaves. PMID- 10380927 TI - Polypeptide flux through bacterial Hsp70: DnaK cooperates with trigger factor in chaperoning nascent chains. AB - A role for DnaK, the major E. coli Hsp70, in chaperoning de novo protein folding has remained elusive. Here we show that under nonstress conditions DnaK transiently associates with a wide variety of nascent and newly synthesized polypeptides, with a preference for chains larger than 30 kDa. Deletion of the nonessential gene encoding trigger factor, a ribosome-associated chaperone, results in a doubling of the fraction of nascent polypeptides interacting with DnaK. Combined deletion of the trigger factor and DnaK genes is lethal under normal growth conditions. These findings indicate important, partially overlapping functions of DnaK and trigger factor in de novo protein folding and explain why the loss of either chaperone can be tolerated by E. coli. PMID- 10380928 TI - Rescue of cyclin D1 deficiency by knockin cyclin E. AB - D-type cyclins and cyclin E represent two very distinct classes of mammalian G1 cyclins. We have generated a mouse strain in which the coding sequences of the cyclin D1 gene (Ccnd1) have been deleted and replaced by those of human cyclin E (CCNE). In the tissues and cells of these mice, the expression pattern of human cyclin E faithfully reproduces that normally associated with mouse cyclin D1. The replacement of cyclin D1 with cyclin E rescues all phenotypic manifestations of cyclin D1 deficiency and restores normal development in cyclin D1-dependent tissues. Thus, cyclin E can functionally replace cyclin D1. Our analyses suggest that cyclin E is the major downstream target of cyclin D1. PMID- 10380929 TI - A striking organization of a large family of human neural cadherin-like cell adhesion genes. AB - We have identified 52 novel human cadherin-like genes organized into three closely linked clusters. Comparison of the genomic DNA sequences with those of representative cDNAs reveals a striking genomic organization similar to that of immunoglobulin and T cell receptor gene clusters. The N-terminal extracellular and transmembrane domains of each cadherin protein are encoded by a distinct and unusually large exon. These exons are organized in a tandem array. By contrast, the C-terminal cytoplasmic domain of each protein is identical and is encoded by three small exons located downstream from the cluster of N-terminal exons. This unusual organization has interesting implications regarding the molecular code required to establish complex networks of neuronal connections in the brain and the mechanisms of cell-specific cadherin-like gene expression. PMID- 10380930 TI - Structure of a heterophilic adhesion complex between the human CD2 and CD58 (LFA 3) counterreceptors. AB - Interaction between CD2 and its counterreceptor, CD58 (LFA-3), on opposing cells optimizes immune recognition, facilitating contacts between helper T lymphocytes and antigen-presenting cells as well as between cytolytic effectors and target cells. Here, we report the crystal structure of the heterophilic adhesion complex between the amino-terminal domains of human CD2 and CD58. A strikingly asymmetric, orthogonal, face-to-face interaction involving the major beta sheets of the respective immunoglobulin-like domains with poor shape complementarity is revealed. In the virtual absence of hydrophobic forces, interdigitating charged amino acid side chains form hydrogen bonds and salt links at the interface (approximately 1200 A2), imparting a high degree of specificity albeit with low affinity (K(D) of approximately microM). These features explain CD2-CD58 dynamic binding, offering insights into interactions of related immunoglobulin superfamily receptors. PMID- 10380931 TI - A structural explanation for the binding of multiple ligands by the alpha-adaptin appendage domain. AB - The alpha subunit of the endocytotic AP2 adaptor complex contains a 30 kDa "appendage" domain, which is joined to the rest of the protein via a flexible linker. The 1.9 A resolution crystal structure of this domain reveals a single binding site for its ligands, which include amphiphysin, Eps15, and epsin. This domain when overexpressed in COS7 fibroblasts is shown to inhibit transferrin uptake, whereas mutants in which interactions with its binding partners are abolished do not. DPF/W motifs present in appendage domain-binding partners are shown to play a crucial role in their interactions with the domain. A single site for binding multiple ligands would allow for temporal and spatial regulation in the recruitment of components of the endocytic machinery. PMID- 10380932 TI - A synthetic peptide inhibitor for alpha-chemokines inhibits the tumour growth and pulmonary metastasis of human melanoma cells in nude mice. AB - Growth-related oncogene-alpha (GROalpha) was first described as an autocrine mitogen and growth factor for melanoma cells. More recent studies show that GROalpha, interleukin-8 (IL-8) and other members of the alpha-chemokine superfamily are also angiogenic. Therefore, we sought to determine if inhibitors of the alpha-chemokine receptor would be effective in inhibiting the tumour growth and pulmonary metastasis of human melanoma cells. We determined that melanocytes and 12 human melanoma cell lines produce both GROalpha and IL-8. The proliferation of A375SM, a highly metastatic cell line, and C8161-C were significantly increased by human recombinant GROalpha and inhibited by anti-human GROalpha monoclonal antibody. Antileukinate, a potent inhibitor of alpha chemokine receptor binding, inhibited the binding of GROalpha to its receptors in melanocytes and all 12 melanoma cell lines tested. Antileukinate also suppressed proliferation of A375SM and C8161-C cells in a dose-dependent manner, and the suppression was not due to cytotoxic effects. Furthermore, continuous administration of antileukinate inhibited the tumour growth and pulmonary metastasis of A375SM cells in athymic BALB/c nude mice. These findings suggest that antileukinate inhibits the growth of melanoma cells by preventing GROalpha from binding to its receptors. This suggests a possible use of alpha-chemokine receptor inhibitors such as antileukinate in the treatment of malignant melanoma. PMID- 10380933 TI - Gamma radiation and MC540 photosensitization of melanoma in the hamster's eye. AB - The purpose of this study was to evaluate the effects of cobalt-60 gamma radiation and argon laser irradiation using injected merocyanine (MC540) as a photosensitizer on pigmented and non-pigmented Bomirski hamster melanomas growing in the eye. The animals were treated with one of four regimens, receiving gamma irradiation only, photosensitization only, a combination of gamma-irradiation and photosensitization, or a combined time-fractionated treatment. Tumours were exposed to laser light 24 h after injection, when the photosensitizing dye concentration was highest. The degree of tissue damage was evaluated by observation of the area for necrosis, interruption of blood circulation, and the shape and dissemination of the tumour cells. Additionally, tumour growth was monitored through the measurement of tumour volume and also calculated from histological cross sections on the assumption that the tumour morphology is hemi ellipsoidal. A single treatment of tumours by a combination of photodynamic therapy and ionizing radiation resulted in an additive effect, inhibiting tumour growth for 2-4 days. A time-fractionated treatment, given four times every 24 h, markedly delayed tumour growth for up to 6 weeks. The results indicate that MC540 mediated photodynamic treatment in combination with gamma-radiation exerts a significant therapeutic effect on a rapidly growing melanoma. PMID- 10380934 TI - Treosulfan is an effective alkylating cytostatic for malignant melanoma in vitro and in vivo. AB - The therapy of metastatic malignant melanoma is limited by poor responses and short overall survival. Thus it remains an important issue to identify and test potential new drugs in this disease. This study was performed to examine the effects of the bifunctional alkylating cytostatic treosulfan in vitro. Using an in vitro microplate ATP bioluminescence tumour chemosensitivity assay (ATP-TCA) five highly chemoresistant melanoma cell lines and melanoma cells freshly isolated from metastases surgically resected from stage IV melanoma patients (n = 10) were incubated with treosulfan. Three cell lines and eight of the 10 tested tumour cells isolated from melanoma metasteses showed tumour growth inhibition >50% after incubation with treosulfan. Therefore, 14 patients with rapidly progressing stage IV malignant melanoma who had been pretreated with at least one standard chemotherapy regimen received treosulfan. In this population of patients with highly refractory advanced melanoma, one complete remission (7.1%), two partial remissions (14.3%) and three cases of stable disease (21.4%) were observed. The median survival time for all the patients measured from the beginning of treosulfan treatment was 9 months, and the median overall survival was 17 months. Except for two patients who developed grade 3 leucopenia, only moderate side effects were observed. Therefore, we conclude that treosulfan was well tolerated in this small series of patients and seems to be a promising alkylating cytostatic for the treatment of metastatic melanoma. Further studies are warranted to test these findings. PMID- 10380935 TI - Normal repair of ultraviolet radiation-induced DNA damage in familial melanoma without CDKN2A or CDK4 gene mutation. AB - Excessive sun exposure and family history are strong risk factors for the development of cutaneous melanoma. Inherited susceptibility to this type of skin cancer could therefore result from constitutively impaired capacity to repair ultraviolet (UV)-induced DNA lesions. While a proportion of familial melanoma kindreds exhibit germline mutations in the cell cycle regulatory gene CDKN2A (p16INK4a) or its protein target, cyclin-dependent kinase 4 (CDK4), the biochemical basis of most familial melanoma is unknown. We have examined lymphoblastoid cell lines from melanoma-affected and unaffected individuals from large hereditary melanoma kindreds which are not attributable to CDKN2A or CDK4 gene mutation. These lines were tested for sensitivity of clonogenic growth to UV radiation and for their ability to repair transfected UV-damaged plasmid templates (host cell reactivation). Two of seven affected-unaffected pairs differed in colony survival after exposure to UVB radiation; however, no significant differences were observed in the host-cell reactivation assays. These results indicate that melanoma susceptibility genes other than CDKN2A and CDK4 do not impair net capacity to repair UV-induced DNA damage. PMID- 10380936 TI - Loss of heterozygosity at chromosome 9p21 (INK4-p14ARF locus): homozygous deletions and mutations in the p16 and p14ARF genes in sporadic primary melanomas. AB - Loss of heterozygosity (LOH) was determined in 45 sporadic primary melanomas at six polymorphic microsatellite markers that flank the INK4a (p16-p14ARF) locus on chromosome 9p21. We also determined allelic loss at two markers on chromosome 9q and two markers at the Rb locus on chromosome 13. Homozygous deletion of the p16 and p14ARF genes was determined by a fluorescent-based quantitative multiplex polymerase chain reaction method. LOH at one or more polymorphic microsatellite markers on locus 9p21 was found in 32 of the melanomas (71%). The highest proportion of LOH was found at markers D9S736 and D9S104, which are telomeric and centromeric to the INK4 locus, respectively. Five melanomas showed LOH at all the analysed markers located on chromosome 9p21. LOH at markers D9S942 and D9S974, which are located close to the p16 and p14ARF genes, was found in 39% and 46% of melanomas, respectively. Analysis of the marker D9S257 on 9q22.1 showed LOH in 13 melanomas (44% of the informative cases). A subset of melanomas with LOH at the INK4 locus also carried inactivating mutations within the p16 coding sequence. Four melanomas carried homozygous deletions at the p16-p14ARF locus. Our results suggest, besides the involvement of the INK4 locus in sporadic melanomas, the possibility of the existence of additional tumour suppressor loci on chromosome 9. PMID- 10380937 TI - Expression of cell cycle regulators in human cutaneous malignant melanoma. AB - We postulate that genes involved in the control of cell proliferation are important determinants of melanoma growth and/or transformation. Using Western blot analysis, we compared the expression of nine key cell cycle regulators in metastatic melanomas with that in benign acquired naevi. Among the cyclin dependent kinases (CDKs) examined, CDK2 was consistently and significantly overexpressed (three- to eight-fold) in metastatic melanomas compared with naevi. CDK1 and CDK4 exhibited no significant difference in expression between benign naevi and metastatic melanomas. CDK6 expression was variable, with four out of 10 metastatic melanomas showing higher expression than naevi. All the cyclins examined, especially cyclins A and D, were expressed more in metastatic melanomas than in naevi. Cyclin E was not detected in benign naevi, but was easily detectable in most of the metastatic melanomas. In addition, there was significantly greater expression of CDC25A, a tyrosine phosphatase that activates CDK kinases, in the metastatic melanomas. Over-expression of CDK2, CDK6, CDC25A and cyclin A was confirmed in melanoma cell lines. These cell cycle regulators may play an important role in melanoma growth and/or transformation. PMID- 10380938 TI - Prognostic significance of serum S100B detection compared with routine blood parameters in advanced metastatic melanoma patients. AB - Recent reports on the use of a quantitative measurement of S100B protein for the detection of metastatic melanoma have yielded promising results. In this study we evaluated 489 serum samples from 64 patients suffering from advanced melanoma (UICC/AJCC stage IV) to compare the sensitivity of a S100B immunoradiometric assay (IRMA) with that of conventional blood parameters as well as other known clinical prognostic factors. In a univariate statistical analysis, gender, bone metastasis, and lactate dehydrogenase and S100B levels in serum samples were found to be significant prognostic markers (P<0.05). The S100B level represented the only relevant independent prognostic marker that was sustained in a multivariate analysis (P = 0.016). Furthermore, we were able to demonstrate that S100B is of relevance irrespective of the specific sites of metastatic involvement. The other laboratory parameters could not match the sensitivity rate of S100B. Overall survival rate was strongly associated with serum S100B values. The results of our study suggest that S100B might be a useful tool as a melanoma marker and an independent prognostic factor in advanced metastatic melanoma. S100B serum detection is likely to be of great interest for the pretreatment stratification and/or monitoring of patients enrolled in clinical studies. PMID- 10380939 TI - Digital videomicroscopy and image analysis with automatic classification for detection of thin melanomas. AB - The aim of our investigation was to evaluate the usefulness of a system composed of a digital videomicroscope equipped with a dedicated program for the quantitative characterization of various parameters of the clinically significant features of pigmented skin lesion (PSL) images, forming the basis for automatic differentiation of naevi and thin melanomas. In total 424 naevi and 37 melanomas (including 23 thinner than 0.75 mm) were considered. All the digital images were acquired, framed and analysed using the DBDermo-MIPS program (Biomedical Engineering Dell'Eva-Burroni), which calculates different parameters related to the geometry, the colour distribution and the internal pattern of the lesion. We also assessed the efficacy of an automatic classifier, trained for 100% sensitivity using a subset of PSL images (59 naevi and 19 melanomas), on a test set including 365 naevi and 18 melanomas thinner than 0.75 mm. Significant differences between values from benign and malignant PSLs were observed for most of the numerical parameters. Values from the training set underwent elaboration by means of multivariate discriminant analysis, enabling the identification of variables that are important for distinguishing between the groups in order to develop a procedure for predicting group membership for new cases (test set) in which group membership is undetermined. Going on the training set data, a threshold score was established, enabling each melanoma to be attributed to the right group. When the same threshold value was employed for discriminating between benign and malignant lesions in the test set, all the melanomas were correctly classified, whereas 30 out of the 365 benign lesions were attributed to the wrong group. Thus the specificity of the system reached 92%, whereas the sensitivity was 100%. Our data suggest that elaboration of videomicroscopic images by means of dedicated software improves diagnostic accuracy for thin melanoma. Since elaboration of an image requires only 60s using our system, all the parameter data are available in real time and can be immediately examined by the classifier, providing an instant aid to clinical diagnosis. PMID- 10380940 TI - Cytokine production by CD4+ T-cells responding to antigen presentation by melanoma cells. AB - Melanoma cells are unusual because, unlike most epithelial tumours, constitutive expression of HLA class II antigens is common. We have previously demonstrated that a peptide-specific CD4+ T-cell clone proliferates briskly in response to peptide and HLA class II expressing melanoma cell lines derived from metastases. Here we demonstrate that these CD4+ T-cells secrete large amounts of interferon gamma (IFNgamma) and interleukin-10 (IL10), and insignificant quantities of IL2 or IL4, in response to peptide presentation by both melanoma and autologous B cells. T-cells produced more IL10 when responding to peptide presentation by melanoma cells compared with B-cells, and less IFNgamma (P<0.01). Addition of IL12 did not alter the cytokines produced but increased the T-cell production of both, especially the production of IL10 in response to peptide presentation by melanoma cells. Our data suggest that differential cytokine production by CD4+ T cells in response to peptide presentation by HLA class II expressing tumour cells may contribute to tolerance to tumour antigens. PMID- 10380941 TI - Is there any relationship between interleukin-6/interleukin-6 receptor modulation and endogenous interleukin-6 release in metastatic malignant melanoma patients treated by biochemotherapy? AB - During recent years it has become clear that the production of most cytokines could play an important role in malignancies. We previously demonstrated that a high endogenous interleukin-6 (IL-6) level is significantly correlated with a high tumour burden and resistance to biochemotherapy in metastatic malignant melanoma patients. However, little is known about the origin of IL-6 and the pattern of IL-6 receptor (IL-6R) expression. In this report, we studied the expression of IL-6R and intracellular IL-6 using flow cytometry in tumour cells provided by fine-needle aspiration of lymph nodes and palpable metastatic lesions from 14 patients refractory to biochemotherapy and six responder patients. Moreover, we established the relationship between these parameters and the serum IL-6 level. Our results demonstrated that, following treatment, the percentage of HMB45-positive (HMB45+) cells expressing functional IL-6R, intracellular IL-6 or both IL-6R and IL-6 significantly decreased in patients refractory to biochemotherapy. In contrast, in responder patients the percentage of HMB45+ cells expressing IL-6R increased and those expressing IL-6 remained stable. Regarding the serum IL-6 level, an 11-fold increase was observed in the patients refractory to biochemotherapy, but only a 1.8-fold increase in the responder patients. In conclusion, in metastatic malignant melanoma patients with a poor prognosis, the endogenous production of IL-6 is concomitant with a decrease in functional IL-6R and intracellular IL-6 expression, suggesting the involvement of an IL-6/IL-6R complex. PMID- 10380942 TI - Clinical and histopathological characteristics in relation to aetiological risk factors in cutaneous melanoma: a population-based study. AB - In this population-based, case-control study from Sweden using data collected from 1988 to 1990, an increased risk of melanoma was associated with the number of sunburns, propensity to freckle, the number of raised naevi and a family history of melanoma. Furthermore, a decreased risk was associated with occupational sun exposure. The purpose of this study was to investigate whether different histopathological features of the melanoma and clinical factors were related to the different aetiological risk factor patterns. All the confirmed primary cutaneous melanomas (n = 366) were included in the study. Both univariate analyses with tests for interaction and multivariate analyses were performed. Patients with melanoma on the trunk and patients with thin melanomas had an excess of close relatives with a history of melanoma (odds ratios [ORs] = 2.7 and 2.3, respectively). A relationship was also seen between melanomas in younger persons and a family history of melanoma (OR = 2.6). The presence of raised naevi on the arm had a tendency to be closer related to melanoma of the nodular type (OR = 4.3) than melanoma of the superficial spreading type (OR = 1.6). Patients with outdoor occupations during summer had a decreased risk of developing melanoma on the extremities. Melanoma diagnosed in patients born before 1939 had an association with sunburns (OR = 1.9) and freckling (OR = 2.0), while melanomas in patients born in 1939 or later were related to a family history of melanoma (OR = 2.2). These results suggest that different histopathological and clinical features of melanoma are associated with different risk factor patterns, which may imply diverging tumour genesis. PMID- 10380943 TI - Cutaneous malignant melanoma and sun exposure in Spain. AB - Cutaneous malignant melanoma has an increasing importance all over the world. However very few epidemiological studies have been published from Spain, and Spanish people have not become aware of the problem. This study was designed to examine sun exposure patterns and other related items among 116 consecutive patients with melanoma and 235 controls. Each subject answered a questionnaire covering the place of residence, sun exposure details and other risk factors, and underwent a skin examination. Continuous sun exposure due to residence or occupation was associated with an odds ratio (OR) of 2.0 (95% confidence interval [CI] = 1.2-3.3). People who lived in the city but spent 50% of their time in rural areas for holidays had an OR of 2.2 (95% CI = 1.3-3.8) when compared with those living in urban and rural areas. The OR for people who sunbathed more than 30 times a year was 1.8 (95% CI = 1.2-2.8), and outdoor leisure time was also associated with melanoma appearance when exposure was greater than 60 units in the last 2 years, with an OR of 3.0 (95% CI = 1.6-5.5); 1 unit is equivalent to total body sun exposure for at least 2 h. These OR estimates were adjusted for age, skin type and the number of naevi. Construction workers (OR = 1.6; 95% CI = 0.5-5.6) had increased risk after adjustment for skin type, age and freckle count (OR = 4.3; 95% CI = 1.8 9.9) or mole count (OR = 2.8; 95% CI = 1.4-5.8). Working as a farmer was a protective factor after adjustment (OR = 0.5; 95% CI = 0.3 0.8). The use of sunscreens was a protective factor against melanoma (OR = 2.6; 95% CI = 1.6-3.6 for non-users). Campaigns should focus on advising people to avoid sun exposure in sunny places and to use sunscreens every time they are exposed to the sun. PMID- 10380944 TI - Neuroelectric assessment of nutrient intake. AB - Electroencephalographic (EEG) activity and auditory event-related brain potentials (ERPs) were assessed in two groups (n = 12 each) of subjects. The 'food-nutrient' group had fasted from the night before and consumed a 500 cal nutrient drink; the 'control' group consumed breakfast but did not consume any nutrients during the recordings. All subjects were assessed every 15 min for six trial blocks at the same time of day, with the fast/nutrient group measured initially before and after consuming the nutrient drink. No effects of the nutrient drink were obtained on the post-stimulus EEG spectral power or mean frequency measures. However, the fast/nutrient group demonstrated less delta, theta, and alpha-1 power than the no-fast/control group. Increases in spectral power were generally observed across trial blocks especially for the alpha and beta bands, and are consistent with increases in arousal level. P300 amplitude was unaffected by the nutrient consumption, but target stimulus N100 amplitude was smaller for the food-nutrient compared to the control group. Taken together, the results suggest that nutrient consumption does not directly affect EEG or cognitive ERP measures. PMID- 10380945 TI - EEG power changes during a multiple level memory retention task. AB - EEG changes related to the amount of information held in memory during a 4-s retention period were studied. The predictability of the amount of information held in memory was varied. In the weighted condition, 60% of the trials contained only one item and the remaining 40% of the trials were evenly distributed between trials containing 3, 5, 7, or 8 items. In the random condition, the levels were equally represented and randomly presented. In the blocked condition the levels were equally represented but presented in five blocks containing only items from one of the levels. Initial widespread decreases in alpha band power were followed by increased activity in all three conditions. The more difficult of the five levels produced decreased alpha activity in more localized posterior left hemisphere sites. This suggests two alpha mechanisms, one associated with task engagement and the other related to the cognitive demands regardless of the presentation context. Theta band power increased over frontal scalp, and to a lesser extent over left parietal and temporal areas and bilateral occipital sites, during only the weighted condition. These changes were uniform over the entire retention period. Beta 2 activity was also influenced by the task difficulty and the time course of the retention period in the two conditions. Beta 2 activity resembled both alpha and theta in that in levels 1, 2 and 3 it acted like alpha with increasing power over time at numerous widespread sites while the higher difficulty levels showed higher power at the beginning of the retention period and then decreased. PMID- 10380946 TI - Validation of a right hemisphere vigilance system as measured by principal component and factor analyzed quantitative electroencephalogram. AB - Arruda and colleagues [Arruda, J.E., Weiler, M.D., Valentino, D.A. et al., 1996. A guide for applying principal-component analysis and confirmatory factor analysis to qEEG data. Int. J. Psychophysiol. 23, 63-81.] recently described seven neurophysiological measures that were previously derived and confirmed using factor analytic procedures and the quantitative electroencephalogram (EEG) sampled from 208 normal controls during an auditory continuous performance test (CPT). The purpose of the present investigation was to further test the validity of these empirically derived measures by examining each measure's relationship with CPT-related declines in performance. Participants were 48 right-handed men (n = 13) and women (n = 35) who reported being free of any neurological condition, birthing complications, or loss of consciousness greater than 2 min. After completing an eyes-closed resting condition, participants performed a 23 min CPT while both quantitative EEG and behavioral performance were measured at 45, 405, 765 and 1125 s into the CPT. Bipolar recordings were gathered using the International 10-20 system, from eight sites: frontal, fronto-temporal, temporal and temporal-occipital. Multivariate and follow-up univariate tests suggest the existence of a neurophysiological system located within the right temporal region that appears essential for the maintenance of a sustained attentional state. If confirmed, the further quantification of this neurocognitive system may prove useful as part of a clinical diagnostic workup. PMID- 10380947 TI - Identifying and reducing noise in psychophysiological recordings. AB - Psychophysiology continues to be a widely used methodology in the study of human behaviour, emotion and cognition. The new researcher is faced with a number of problems in the recording process since the desired physiological signal must be isolated from a variety of noise sources. Precautions and strategies that can be implemented in setting up the recording equipment and isolating the subject from interference are described. There are also a number of software techniques that can be applied to improve signal quality after the data have been acquired. An overview is provided of hardware and software methods used to maximise the signal quality. PMID- 10380948 TI - Theta synchronization during sustained anticipatory attention in infants over the second half of the first year of life. AB - The neurophysiological basis of attention control has been investigated in infants during the second half of the first year of life. The marked improvement of voluntary control of attention and action is known to occur during this age period. EEG was registered in 60 infants aged 8-11 months under three experimental conditions: (1) attention to an object in the visual field (externally controlled attention or the 'baseline'); (2) anticipation of the person in the peek-a-boo game (internally controlled attention); and (3) attention to the 'reappeared' person in the peek-a-boo game ('control' condition). Spectral analysis of the data revealed sharp increase of the EEG theta activity (3.6-6.0 Hz) during internally controlled attention as compared to the 'baseline' and the 'control' conditions. The theta1 (3.6-4.8 Hz) increase was maximal at frontal electrode sites. The reactivity of the frontal theta1 during internally controlled attention differentiated subjects with different ability to maintain this type of attention. The theta2 (5.2-6.0 Hz) reactivity was maximal at right temporal electrode site and did not depend on the ability to maintain anticipatory attention. The data point to different functional significance of theta1 and theta2 rhythms in infants. It was suggested that the frontal theta1 synchronization in infants reflects activity of the anterior attention system subserving executive control of attention. The ability to maintain anticipatory attention increased, whereas the frontal theta1 synchronization decreased during the studied age period. There was the direct relationship between frontal theta1 synchronization and persistence of internally controlled attention in 8-month olds. On the contrary, at 9 and 10 months, these variables were inversely related. There was no link between theta1 reactivity and persistence of anticipatory attention in 11-month-olds. It was suggested that the age-dependant dynamic of the relationship between frontal theta1 reactivity and attention behaviour reflects the maturational shift in the functioning of anterior attention system. The shift leads to more economic and more efficient functioning of this system. PMID- 10380950 TI - The so-called 'mind/brain dualism', the 'bi-cameral mind' and 'the double brain'. PMID- 10380949 TI - The effects of a 20-min nap at noon on sleepiness, performance and EEG activity. AB - The prophylactic effects of a 20-min nap at noon on afternoon sleepiness were studied. Ten young adults who had normal sleep-wake habits without habitual daytime napping were subjected to nap and no-nap conditions at an interval of 1 week. After a nocturnal sleep recording (00.00-08.00 h), their EEG recordings during relaxed wakefulness, mood, performance, and self-ratings of performance level were measured every 20 min from 10.00 h to 18.00 h. For the nap condition, they went to bed at 12.20 h and were awakened when 20 min had elapsed from the onset of sleep stage 1. For the no-nap condition, they rested without sleeping by sitting on a semi-reclining chair. The nap did not improve task performance, however, it improved volition and the self-rating of task performance. It also suppressed subjective sleepiness and attenuated eyes-opened EEG alpha activities. The results suggest that a 20-min nap at noon had partial positive effects on the maintenance of the daytime arousal level. PMID- 10380952 TI - fMRI evidence for an inverted face representation in human somatosensory cortex. AB - We provide evidence that the face component of the somatosensory homunculus is actually upside down rather than right-side up along the central sulcus of the human brain. We pneumatically stimulated the forehead or chin of neurologically intact humans while acquiring fMRI images of somatosensory cortex. During forehead stimulation cortical regions along relatively inferior portions of the postcentral gyrus were most active whereas during chin stimulation relatively superior regions were most active. These data are consistent with an inverted face representation along the central sulcus of the human brain. PMID- 10380951 TI - The use of diffusion tensor imaging in quantifying the effect of dexamethasone on brain tumours. AB - The role of dexamethasone in the treatment of patients with brain tumours remains poorly understood. In this study the self-diffusion parameters of extracellular water within primary intracranial tumours and peritumoural tissue, and their response to dexamethasone, have been measured using MR diffusion tensor imaging. Maps of the mean diffusivity and two measures of diffusion anisotropy were obtained from six patients before, and typically 48-72h after, commencing dexamethasone treatment. A significant decrease in of either tumour (p < 0.02) or oedematous brain (p < 0.04) was observed in three patients. No significant changes were noted in either of the two calculated diffusion anisotropy indices before and after steroid treatment in any of the six patients. PMID- 10380953 TI - Non-linear EEG dynamic changes and their probable relation to voluntary movement organization. AB - This study was undertaken to analyze systematically the non-linear dynamic changes of EEG activity accompanying slow goal-directed voluntary movements, using three non-linear characteristics (NC): point-wise correlation dimension, Kolmogorov entropy and largest Lyapunov exponents as functions of time. NC indicated transitions with non-linear properties (NT). A significant difference between times of appearance of the NT with respect to the electrode position was established: before the movement onset, NT appeared first in contralateral and midline areas including frontal, sensorimotor and parietal cortices. Before target reaching, NT appeared first in the contralateral sensorimotor area, and evolved ipsilaterally. The results suggest that the NT could be regarded as precursors of higher functional coupling between cortical areas involved in voluntary movement organization. PMID- 10380954 TI - Periaqueductal gray matter glutamate and GABA decrease following subcutaneous formalin injection in rat. AB - Glutamate and GABA are important nociception modulating transmitters in specific brain regions, i.e. the spinal cord, the thalamic nuclei and the periaqueductal gray (PAG). However, quantitative and topographical changes in glutamate and GABA release in these brain regions during peripheral inflammation episodes have not been characterized in awake animals. To address this issue, an in vivo microdialysis study was carried out in freely moving rats in order to analyze PAG extracellular glutamate and GABA concentrations following unilateral formalin injection into the dorsal skin of the right hind-paw. Both glutamate and GABA release decreased after the injection of formalin during phase I and phase II of hyperalgesia. Because naloxone prevented the decrease of GABA and glutamate release induced by formalin, this study shows that, in vivo, a nociceptive stimulation may activate opioidergic fibres into the PAG. The increased release of endogenous opioids may, in turn, inhibit the activity of the GABAergic neurons (i.e. opioid disinhibition). Formalin injection also decreased extracellular glutamate concentration. However, we found that intra-PAG perfusion with tetrodotoxin only decreased GABA, but not glutamate dialysate values. Although it should be reasonable to speculate that opioids also inhibit glutamate fibres, further investigation is needed to clarify whether or not the dialysate glutamate we measured reflects change in the metabolism or neurotransmitter pool of this amino acid. PMID- 10380955 TI - A novel candidate presenilin-1 interacting protein containing tetratricopeptide repeats. AB - The yeast two-hybrid system, immunofluorescence and co-immunoprecipitation techniques were used to identify a novel candidate protein with which presenilin 1 (PS-1) interacts. This interacting protein, the gene of which is encoded on chromosome 16, contains two tetratricopeptide repeats (TPR) that are known to mediate interactions between proteins, appears to be primarily localized to the cytoplasm of transfected HEK293 cells, and is expressed in brain. Preliminary yeast two-hybrid data suggests this candidate may interact with both heat shock protein-90 and heat shock protein-70 and thus may be a novel member of TPR containing proteins which interact with this complex. PMID- 10380956 TI - Effect of induced tolerance on biochemical disturbances in hippocampal slices from the gerbil during and after oxygen/glucose deprivation. AB - To evaluate whether the state of tolerance is stable enough to be studied under in vitro conditions after induction by ischemic preconditioning in vivo, metabolic disturbances of hippocampal slices prepared from control and preconditioned gerbils were evaluated during and after oxygen/glucose deprivation (OGD). Slices were subjected to 5, 10 or 15 min OGD with or without 2h recovery. During the state of metabolic stress, changes in energy metabolism were identical in slices taken from control and preconditioned gerbils. Following OGD, however, recovery of protein synthesis was significantly improved in hippocampal slices of preconditioned animals, indicating that the effect of preconditioning on metabolic disturbances induced by transient OGD in vitro or transient ischemia in vivo is similar. It is suggested that the hippocampal slice preparation is an in vitro model suitable for the study of basic mechanisms underlying the induction of tolerance in vivo. PMID- 10380957 TI - Axonal neurofilaments are resistant to calpain-mediated degradation in the WLD(S) mouse. AB - The biological basis for the phenotype of delayed Wallerian degeneration in the WLDs mouse has yet to be elucidated, although it is known that the characteristic is intrinsic to the axon. Previous data suggested that nerves from the WLD(S) are relatively resistant to proteolytic degradation. We investigated the time-course of neurofilament degradation in response to addition of the calcium-activated protease m-calpain, comparing nerves from WLD(S) and wild-type mice. During 10 min of in vitro proteolysis, neurofilaments from the WLD(S) were consistently slower to degrade than were neurofilaments from wild-type mice. Direct comparisons were performed on Western blots, with statistically significant differences in neurofilament immunoreactivity at 2, 4, and 6 min of reaction time (p < 0.01). These findings suggest that the mutation leading to the WLD(S) phenotype may affect the proteolytic interaction between calpain and neurofilaments. PMID- 10380958 TI - Cation selective channels formed by a C-terminal fragment of beta-amyloid precursor protein. AB - The C-terminal 105 amino acid fragment of beta-amyloid precursor protein (CT105) is highly neurotoxic. To obtain insights into its cytotoxic effect, we examined the ionophoric effects of CT105 (10-1000 nM) on artificial lipid membranes. Macroscopic membrane conductance increased with CT105 concentration and its ionophoric effect was comparable to that of amyloid beta protein. The mean unitary conductance of CT105-induced channels was 120 pS and open-state probability was close to 1 at voltages from -80 to +80 mV. CT105induced channels were selective to cations (PK/ P(Cl) = 10.2), being most selective to Ca2+. These findings suggest that CT105 can cause direct neurotoxic effects by forming Ca2+ permeable cation channels on neuronal membranes. PMID- 10380959 TI - GDNF, RET and GFRalpha-1-3 mRNA expression in the developing human spinal cord and ganglia. AB - The identification of endogenous neurotrophic factors and their receptors in human spinal cord is important not only to understand development, but also in the consideration of possible future therapies for neurodegenerative disorders and trauma. Using in situ hybridization, the expression of glial cell line derived neurotrophic factor (GDNF), neurturin (NTN), persephin (PSP), GFRalpha-1, GFRalpha-2, GFRalpha-3 and RET mRNA in human fetal spinal cord was studied. Strong GDNF mRNA hybridization signal, presumably restricted to Clarke's nucleus, was detected in the thoracic spinal cord. mRNA encoding GFRalpha-1 was expressed in the entire spinal cord gray matter with particularly high expression in the ventral horn. GFRbeta-1 was also expressed more weakly in dorsal root ganglia. NTN and persephin mRNA were not detected in either the fetal spinal cord or the dorsal root ganglia. mRNA coding for GFRalpha-2, however, was found in most cells of the spinal cord gray matter. A strong expression of GFRalpha-3 mRNA was detected in dorsal root ganglia cells and Schwann cells. The transducing receptor RET was expressed strongly in motorneurons and dorsal root ganglion neurons. We conclude that basic features concerning the role of the GDNF family of ligands and their receptors revealed in rodents applies to humans. PMID- 10380960 TI - Postsynaptic 5-HT1A receptors control 5-HT release in the rat medial prefrontal cortex. AB - 5-HTt1A receptor agonists reduce the neuronal release of 5-hydroxytryptamine (5 HT) by activation of raphe 5-HT1A autoreceptors. Using in vivo microdialysis in unanesthetized rats, we show that the local application of the selective 5-HT1A receptor agonist 8-OH-DPAT decreased the 5-HT output to approximately 50% of controls in medial prefrontal cortex (mPFC) but not in dorsal hippocampus. The decrease in 5-HT output was counteracted by the concurrent application of the selective 5-HT1A receptor antagonist WAY-100635. This agent also reversed the decrease in 5-HT output elicited by the novel 5-HT1A receptor agonist BAY x 3702 (30 microM) in mPFC and dorsal raphe nucleus. These results indicate that postsynaptic 5-HT1A receptors in mPFC also participate in the control of serotonergic activity. PMID- 10380961 TI - Transneuronal degeneration of retinal ganglion cells in early hemispherectomized monkeys. AB - Transneuronal retrograde cell changes in the retina of the primate have been well documented after lesions to striate cortex, but little is known about the effects of hemispherectomy, a surgical procedure used in humans for the treatment of intractable epilepsy. In order to follow the time course of this degenerative process, we examined the retinae of six monkeys who underwent a total hemispherectomy at various postnatal ages with a survival period of 4 years. We demonstrate that transneuronal retrograde degeneration in the retina following hemispherectomy is inversely correlated with age at the time of the lesion. This degeneration is maximal when the lesion is induced within the first 4-6 months of life and less pronounced from 8 months to adulthood. PMID- 10380962 TI - Differential expansion of neural projection systems in primate brain evolution. AB - Whole brain MRI scans from 11 primate species (43 individuals) spanning more than a 50-fold range in brain volume were used to determine whether the corpus callosum keeps pace with the growth of the forebrain among living anthropoid primates. Interhemispheric connectivity via the corpus callosum and anterior commissure was reduced in larger primate brains, whereas intrahemispheric connectivity was augmented. We also show that the splenium constitutes an increasing proportion of callosal area with increasing brain size. This may function to maintain rapid integration of the left and right visual space as brain size increases. These results indicate that the evolution of larger brain size in primates results in increasingly independent hemispheres. PMID- 10380964 TI - Monkey globus pallidus external segment neurons projecting to the neostriatum. AB - Experiments were performed to assess the number and parvalbumin (PV) immunoreactivity of neurons participating in the pallidostriatal projection in macaque monkeys. Injection of WGA-HRP into the right caudate nucleus and the left putamen of a Macaca mulatta and a M. fuscata labeled a large number of the globus pallidus external segment (GPe) neurons. Counting neurons labeled with WGA-HRP and those stained with neuronal markers indicated that approximately 30% of GPe neurons project to neostriatum. Approximately 2/3 of the pallidostriatal neurons are PV-immunoreactive. This study revealed that a significant number of primate GPe PV immunoreactive neurons project to the neostriatum, and suggest that the pallidostriatal projection should be taken into account in the analysis of functional roles of the basal ganglia circuitry. PMID- 10380963 TI - A cyclooxygenase-2 inhibitor attenuates white matter damage in chronic cerebral ischemia. AB - The effects of nimesulide, a cyclooxygenase-2 inhibitor, were examined during chronic cerebral hypoperfusion. After bilateral ligation of the common carotid arteries in 30 rats, 21 received dosages of 2 or 5 mg/kg nimesulide daily and nine received vehicle daily for 14 days. The serum was then analyzed biochemically, and pathological changes were estimated in the white matter by the emergence of major histocompatibility complex (MHC) antigen-immunoreactive activated microglia and white matter lesions. In the vehicle-treated animals, activated microglia and white matter lesions were observed. Following treatment with either 2 or 5mg/kg nimesulide, the magnitude of these changes was reduced (p < 0.001) without significant side effects. These results indicate a potential use for cyclooxygenase-2 inhibitors in cerebrovascular disease. PMID- 10380965 TI - Event-related fMRI analysis of the cerebral circuit for number comparison. AB - Cerebral activity during number comparison was studied with functional magnetic resonance imaging using an event-related design. We identified an extended network of task-related areas that showed a phasic activation following each trial, including anterior cingulate, bilateral sensorimotor areas, inferior occipito-temporal cortices, posterior parietal cortices, inferior and dorsolateral prefrontal cortices, and thalami. We then tested which of these areas were affected by number notation, numerical distance and response side, three variables that specifically target processes of visual identification, quantity manipulation and motor response in a serial-stage model of the number comparison task. Our results confirm the role of the right fusiform gyrus in digit identification processes, and of the inferior parietal lobule in the internal manipulation of numerical quantities. PMID- 10380966 TI - Aberrant plasticity in Alzheimer's disease. AB - Alzheimer's disease (AD) is a region-specific degenerative disease that mainly impairs the temporal and parietal lobes, with preservation of the frontal lobes until advanced stages of disease. Since [11C]diacylglycerol PET facilitates examination of loci exhibiting plasticity, it was performed in eight patients with AD and six age-matched normal control subjects to evaluate frontal lobe function. [11C]Diacylglycerol tomograms obtained from patients with AD demonstrated strong spotty incorporation of [11C]diacylglycerol mainly in the frontal association areas. However, [18F]fluorodeoxyglucose tomograms exhibited region-specific findings such as decreased CMRGIc in the parietotemporal association areas, which are involved in impairment of cognitive function. The strong spotty incorporation of [11C]diacylglycerol suggested a compensatory plastic process in the frontal lobes in response to involvement by Alzheimer's disease of the posterior association areas. PMID- 10380968 TI - Alzheimer's disease may not be a spirochetosis. AB - It has been reported previously that spirochetes could be one of the causes of Alzheimer's disease (AD). In this study, we have attempted to reproduce these findings by examining fresh blood samples from 22 patients diagnosed with early stage (n = 16) and late stage (n = 6) AD. The patients were participants in a clinical drug trial. Fresh necropsy brain cortical specimens from AD patients (n = 7) were also examined. Spirochetes were observed microscopically in the blood of only one late-stage AD patient. None of the brain tissues showed the presence of spirochetes. Our results suggest that spirochetes are probably not associated with AD. PMID- 10380967 TI - An antisense oligonucleotide to brain-derived neurotrophic factor delays postural compensation following unilateral labyrinthectomy in guinea pig. AB - An antisense oligonucleotide to brain-derived neurotrophic factor (BDNF) was delivered by osmotic mini-pump at a 1 mM concentration via a cannula into the ipsilateral vestibular nucleus complex from 15 to 56h following unilateral labyrinthectomy in guinea pigs. Compared with the control groups, vestibular compensation of roll head tilt was significantly delayed (p < 0.05), while compensation of spontaneous nystagmus and yaw head tilt was unaffected. These results suggest that neurotrophins such as BDNF may be involved in specific aspects of the vestibular compensation process. PMID- 10380969 TI - Some membrane property changes following axotomy in A delta-type DRG cells are related to cold allodynia in rat. AB - Numerous studies have suggested that changes in electrophysiological properties of primary sensory neurons after axonal injury contribute to the generation of neuropathic pain. Presently, however, it is unclear which of the changes is important. To address this issue, we performed behavioral and electrophysiological experiments in a double-blind fashion; we made intracellular recordings in the S1 dorsal root ganglia excised from rats exhibiting cold allodynia behavior after chronic S1 spinal nerve transaction (allodynia-positive group) and from rats lacking such behavior after the same nerve injury (allodynia negative group) or sham injury (sham group). In this study, we sought which of the membrane property changes produced by the spinal nerve injury in each of C-, Adelta- and Aalpha/beta-cell populations was unique to the allodynia-positive group. Analyses of our data revealed that only some changes in Adelta-cells (e.g. the decrease in resting membrane potential and in the threshold of central process) were more pronounced in or unique to the allodynia-positive group. We concluded that certain membrane property changes in the somata and dorsal root axons of Adelta-cells might be important in the generation of cold allodynia. PMID- 10380970 TI - Increased [125I]sulpiride binding in the subthalamic nucleus of rats with nigrostriatal lesions. AB - Subthalamic nucleus (STN) hyperactivity follows lesions of mesencephalic dopaminergic neurons in animal models of Parkinson's disease. The mechanism leading to sustained STN hyperactivity in parkinsonism is not well understood, but it seems not to depend on the integrity of striato-pallido-subthalamic connections (the so called indirect pathway). Sustained STN hyperactivity could result from the loss of the direct dopaminergic innervation of the STN. Here we report increased [125I]sulpiride binding in the STN of rats with 6 hydroxydopamine (6-OHDA) lesions of mesencephalic dopaminergic neurons. Furthermore, we found that chronic oral treatment with levodopa reverted the lesion-induced increase in [125I]sulpiride binding. Our results demonstrate that most STN D2-class dopamine receptors are postsynaptic to afferent dopaminergic fibers. Furthermore, they suggest that alterations of local STN dopaminergic mechanisms could play a role in the pathophysiology of parkinsonism and mediate the therapeutic/adverse effects of chronic levodopa administration. PMID- 10380971 TI - Alpha2-macroglobulin polymorphisms in Alzheimer's disease and dementia with Lewy bodies. AB - Dementia with Lewy bodies (DLB) is the second most common cause of dementia in the elderly after Alzheimer's disease (AD). The apolipoprotein E gene (APOE) is a major risk factor, but can only account for approximately 50% of AD cases. Whole genome scanning in late-onset AD families has suggested that a locus on chromosome 12 may contribute significantly to disease development. Recently the alpha2-macroglobulin gene (A2M) on chromosome 12 has been suggested as a candidate locus for AD. We therefore determined the influence of two polymorphisms in A2M, a pentanucleotide deletion 5' to the bait domain exon, and a valine to isoleucine polymorphism in the thiolester site of the protein, in AD and DLB cohorts. No evidence was observed for an association between the thiolester or deletion polymorphisms and AD or DLB alone or when accounting for the APOE epsilon4 allele. We did, however, identify a non-significant excess of deletion homozygotes in the AD and DLB groups. This genotype accounted for 4% of disease cases but was absent in the control population. Given that the A2M deletion polymorphism is non-functional, the chromosome 12 AD/DLB locus may be situated elsewhere and not with these A2M polymorphisms. PMID- 10380972 TI - A new human experimental pain model: the heat/capsaicin sensitization model. AB - The heat/capsaicin sensitization model is a new human experimental pain model that synergistically combines non-invasive physical and chemical methods of nociceptor stimulation to produce stable and long-lasting hyperalgesia with a low potential for skin injury. In 10 healthy volunteers the forearm was stimulated with a 45 degrees C thermode for 5 min to produce an area of secondary hyperalgesia. Applying capsaicin cream for 30 min further expanded the area of secondary hyperalgesia. Periodically heating the treated skin with a previously non-painful temperature of 40 degrees C re-kindled the sensitization enough to maintain stable areas of secondary hyperalgesia for 4h. The evoked pain was moderate and well tolerated. The heat/capsaicin sensitization model should be well suited for studying pain mechanisms and testing new analgesics. PMID- 10380973 TI - Ketogenic diet reduces spontaneous seizures and mossy fiber sprouting in the kainic acid model. AB - The high fat, low carbohydrate, low protein ketogenic diet (KD) has been used to control refractory epilepsy in children since 1920, although its mechanism of action is unknown. Previous animal studies have shown that the KD can increase acute seizure threshold, but the effect of the KD on the process of epileptogenesis has not been studied. We tested the effect of an experimental KD on epileptogenesis in adult rats using the kainic acid (KA) model. P54 rats underwent KA-induced status epilepticus, followed by assignment to a control diet or a KD consisting of (by weight), 14% protein, 70% fat and no carbohydrate. KD fed animals tolerated the diet and maintained ketosis. KD-fed rats had significantly fewer and briefer spontaneous recurrent seizures and less supragranular mossy fiber sprouting, although the degree of hippocampal pyramidal cell damage was similar in both groups. These results provide the first evidence that the KD retards epileptogenesis in an experimental model. PMID- 10380974 TI - ERP correlates of phoneme perception in speech and sound contexts. AB - To address the question of the existence of a phonetic module for speech perception, event-related potentials were recorded using a 32 channel system in subjects performing a detection task where the target was the ambiguous, noise like phoneme /f/ presented either among syllables (speech context) or among environmental sounds (non-speech context). Significant context effects were observed on the N2/P3 complex elicited by the target. In particular, a well localized N2b (250-280 ms) appeared at the left temporoparietal sites on the difference wave between contexts as the result of an enhanced negativity when the target was presented among non-speech stimuli. These findings suggest the involvement of the left temporoparietal region in autonomous, modular processes of speech perception. PMID- 10380975 TI - Evidence for increased GDNF signaling in aged sensory and motor neurons. AB - Several lines of evidence suggest that attenuated neurotrophin signaling may account for some of the aging-related phenotypic changes observed in motor and sensory neurons. Glial-derived neurotrophic factor (GDNF) signals through the GFRalpha-1-RET receptor complex and has trophic effects on both primary sensory neurons and, in particular, motoneurons. In this study we provide evidence using RT-PCR that GDNF, but not neurturin, is strongly up-regulated in target muscles (800%) and to a lesser extent also in peripheral supportive tissues. Results here, and in an earlier study, show that the up-regulation of GDNF in target and supportive tissues parallels an increased neuronal expression of the cognate receptors. Increased GDNF signaling may explain some of the phenotypic characteristics of aging sensory and motoneurons. PMID- 10380976 TI - Identification of human, rat and mouse nocistatin in brain and human nocistatin in brain and human cerebrospinal fluid. AB - Nocistatin was recently isolated from bovine brain and shown to block hyperalgesia and allodynia induced by nociceptin and prostaglandin (PG) E2. The counterparts of human, rat and mouse are deduced from their precursor prepronociceptin to be 30, 35, and 41 residue peptide respectively. To identify these mature forms of nocistatin, three peptides were synthesized and a detection program for nocistatin was developed, using high pressure liquid chromatography (HPLC) along with specific radioimmunoassay (RIA). Nocistatin extracted from human, rat and mouse brain were subjected to HPLC and nocistatin-like immunoreactivity (NST-IR) was determined. All three species showed two NST-IR peaks, one of which coincided with that of the corresponding putative nocistatin. The same NST-IR was also detected in human cerebrospinal fluid (CSF). PMID- 10380977 TI - Increase of cellular hypoxic tolerance by erythromycin and other antibiotics. AB - Antibiotics are used extensively, but in addition to their anti-infectious effects some inhibit cellular energy metabolism. We investigated hypoxic tolerance following in vivo pretreatment with erythromycin and kanamycin, or in vitro pretreatment with ampicillin. Recovery of the CA1 population spike amplitude in hippocampal slices upon 15 min hypoxia improved time-dependently following single i.p. in vivo pretreatment with erythromycin (maximum at 6 h: recovery 90+/-7% (mean s.d.) vs 30% in untreated controls; p<0.01). The hypoxia induced increase in NADH was smaller in slices that recovered from hypoxia. We conclude that antibiotics increase cellular hypoxic tolerance to a varying extent. Use of antibiotics in experimental studies may, therefore, distort conclusions about hypoxic sensitivity and confounding mechanisms. In contrast, antibiotics may provide an effective strategy to induce chemical preconditioning in humans. PMID- 10380978 TI - Temporal discrimination of somesthetic stimuli is impaired in dystonic patients. AB - Clinical and experimental evidence documents abnormal somatosensory functions in dystonia. Despite the fact that somatosensory processing is inherently temporal, mainly spatial aspects of somatosensory functions have so far been assessed in dystonic patients. Seven patients with idiopathic dystonia and nine healthy controls were given pairs of non-noxious electrical stimuli separated by different time intervals and asked to report if they perceived single or double stimuli. Somesthetic temporal discrimination thresholds (STDT) were obtained by computing the shortest time interval at which stimuli, applied to the left or the right hand, were perceived as separate. STDT were significantly higher in dystonic than in controls thus showing for the first time that temporal and not only spatial somatosensory processing is altered in dystonia. PMID- 10380979 TI - Grafts of meningeal fibroblasts in adult rat spinal cord lesion promote axonal regrowth. AB - We have studied the morphological consequences of implantation into the injured adult rat spinal cord of fibroblasts derived from the meninges overlying the cerebral cortex. Our initial objective was to reproduce the well known post traumatic fibroadhesive scar observed in the clinical situation. One month after implantation, instead of having formed a fibroadhesive scar, fibroblasts had promoted the regeneration of peptidergic axons originating from dorsal root afferents and, to a lesser extent, of supraspinal serotonergic fibers at the periphery of the grafts. Using RT-PCR we were able to identify in cultures of meningeal-derived fibroblasts mRNAs for beta-NGF, NT3, aFGF and bFGF, which suggests that the promoting effect on axonal regeneration of these cells is at least in part due to their capacity to synthesize neurotrophic factors. PMID- 10380980 TI - Rotation-induced conditioned rejection in the taste reactivity test. AB - The taste reactivity test was used to evaluate the ability of motion sickness to produce conditioned rejection reactions, a putative measure of nausea in rats. Following three conditioning trials, rats displayed conditioned rejection reactions during an intraoral infusion of a rotation-paired saccharin solution. This is the first demonstration of conditioned rejection produced with a non pharmacological emetic agent and provides support that the conditioned rejection reaction may serve as a rat model of nausea. PMID- 10380981 TI - Alanine-23 of transducin alpha subunit is involved in defining the affinity for betagammma complex. AB - Transducin alpha subunit (alpha(t)1) shows extraordinarily high affinity to G protein betagamma complex. One of the betagamma-binding regions on alphat1 is the amino-terminal helix. Alanine-23 is uniquely found on alphat1 but not other members of the Gi-subfamily. Mutation of alanine-23 into serine reduced the ability of alpha(t)1 to sequester betagamma-mediated stimulation of type II adenylyl cyclase. The functional impairment is independent to the protein expression levels. Molecular modeling indicated that the hydrophobic interaction between the side chains of alanine-23 of alpha(t)1 and leucine-55 of the beta1 subunit could be disrupted by the introduction of a hydroxyl group. This study showed that alanine-23 of alpha(t)1 is probably involved in defining its affinity for the betagamma complex. PMID- 10380982 TI - Binding visual features during high-rate serial presentation. AB - The neural mechanism supporting performance during single and feature conjunction detection was investigated using event-related brain potentials. In different blocks of trials, participants responded to visual targets defined by one of two colors, one of two orientations, or both color and orientation. Participants were faster and more accurate in detecting targets defined by a single feature than for targets defined by a conjunction of features. Compared with the single feature conditions, conjunction targets were associated with enhanced negativity between 230 and 270 ms post-stimulus and showed a delayed P3 latency. The relative timing of feature specific attention effects isolated in difference potential shows that feature conjunction occurs concurrently with the analysis of single features. PMID- 10380983 TI - Down-regulation of ACTH and glucocorticoid receptor immunoreactivity in hypothalamic arcuate neurons after adrenalectomy in the rat. AB - The expression of glucocorticoid receptor (GR) in rat adrenocorticotropin (ACTH) containing neurons in rat brain was immunohistochemically investigated. ACTH containing cell bodies were found mainly in the arcuate nucleus. Most of these neurons exhibited GR immunoreactivities in their nuclei. ACTH-containing nerve fibers were distributed in the bed nucleus of the stria terminalis, periventricular nucleus, retrochiasmatic nucleus, parvocellular part of paraventricular nucleus and dorsomedial hypothalamic nucleus. After adrenalectomy there was a marked decrease of ACTH immunoreactivity, as well as GR immunoreactivity, in neurons of the arcuate nucleus, but ACTH immunoreactivity in the fibers was not affected. These results indicate that glucocorticoids up regulate ACTH and GR production in hypothalamic arcuate neurons, but that glucocorticoid-induced changes could be delayed in the fibers derived from these neurons. PMID- 10380985 TI - Noradrenaline release in the locus coeruleus of conscious rats is triggered by drugs, stress and blood pressure changes. AB - The in vivo release of noradrenaline (NA) in the locus coeruleus (LC) of conscious rats was enhanced by local superfusion of pargyline, idazoxan, bicuculline, AMPA as well as by experimentally induced hypotension. Noise stress considerably enhanced NA release in the LC and this response was promoted after local alpha2-adrenoceptor blockade by idazoxan. Air jet stress and noise stress elicited comparable increases in NA release in the LC and the simultaneously superfused amygdala. The NA responses in both areas did not change during a second exposure to each of the stressors. It is concluded that NA release at the somatodendritic level of LC neurons is triggered by high LC activity and most likely serves to limit LC activation to excitatory stimuli by feedback inhibition via alpha2-adrenoceptors. PMID- 10380984 TI - Uridine activates fast transmembrane Ca2+ ion fluxes in rat brain homogenates. AB - The excitatory actions of the pyrimidine nucleoside uridine, and the nucleotides UDP and UTP, as well as the purine nucleotide ATP, were studied by fluorescent labeling of Ca2+ and K+ ion fluxes on the time scale of 0.04 ms to 10s in resealed plasmalemma fragments and nerve endings from the rat cerebral cortex. Two phases of Ca2+ ion influx with onsets of a few milliseconds and a few hundred milliseconds, showing different concentration dependencies, agonist sequences and subcellular localizations were distinguishable. [3H]Uridine identified high (K(D) approximately 15 nM) and low affinity (K(D)approximately 1 microM) specific binding sites in purified synaptosomal membranes. Labeled uridine taken up by synaptosomes in a dipyridamole-sensitive process was released by depolarization (1 mM 4-aminopyridine). Taken together, these results may qualify uridine as a neurotransmitter. PMID- 10380986 TI - Association of the androgen receptor gene (AR) with ADHD and conduct disorder. AB - The male predominance of externalizing behaviors suggests that the X-linked androgen gene might be involved. Since the shorter alleles of the CAG and GGC polymorphisms of the AR gene are associated with increased gene expression we sought to determine whether they were also associated with externalizing behaviors. We examined 302 subjects consisting of Tourette syndrome probands and controls. ANOVA showed a significant association between the AR haplotypes and ADHD (p < 0.0001), conduct disorder (CD; p < 0.017), and oppositional defiant disorder (ODD; p < 0.004) with the lowest scores in those with the longer alleles at both polymorphisms. These results suggest that genetic variation at the human AR gene plays a role in human externalizing disorders. PMID- 10380987 TI - Ischemia induces a selective biphasic response in brain mitochondrial mRNA levels. AB - We determined the independent effects of hypoxia, glucose deprivation and ischemia (hypoxia plus glucose deprivation) on steady-state levels of mRNA coding for specific nuclear and mitochondrially encoded enzymes of oxidative metabolism in cultured rat neurons and glia. Neither hypoxia nor low glucose alone changed steady-state message levels for any transcript. However, ischemia induced a biphasic effect on mitochondrially encoded transcripts for cytochrome oxidase subunit two (CO2) and the subunits 8 and 6 of ATPase (A 8/6), initially decreasing and then increasing mRNA levels to or above the levels recorded prior to ischemia. In contrast, three nuclear encoded transcripts for mitochondrial proteins were decreased by ischemia. These data demonstrate a lack of coordination between the expression of nuclear and mitochondrial genes in the initial response to ischemia and suggest that a selective, primary reaction to brain cell insults exists within the mitochondrion. PMID- 10380988 TI - Functional expression of metabotropic glutamate receptor type 5 in rat pinealocytes. AB - Mammalian pinealocytes, endocrine cells for melatonin, express class II metabotropic glutamate receptors (mGluRs), which are involved in negative regulation of melatonin synthesis through an inhibitory cAMP cascade. We investigated whether mGluRs other than class II receptors are expressed in rat pinealocytes. RT-PCR analysis and Northern blotting indicated that the mRNA of mGluR5, a class I receptor, was present in pineal glands. Quisqualate and 1 aminocyclopentane-1,3-dicarboxylate (1S,3R-ACPD), class I receptor agonists, increased the intracellular [Ca2+] of fura-2 loaded cultured pinealocytes in the absence of extracellular Ca2+ which is blocked by methylcarboxyphenylglycine, a class I receptor antagonist. These results suggest that mGluR5 is functionally expressed in pinealocytes and triggers Ca2+ efflux from intracellular stores. PMID- 10380989 TI - Human brain specialization for phonetic attention. AB - The effects of auditory selective attention on event related potentials (ERPs) to speech sounds were examined in subjects attending to vowel-consonant-vowels (VCVs) in one ear while ignoring VCVs in the opposite ear. In one condition, subjects discriminated phonetic changes in the VC, CV, or both formant-transition regions. In another condition, they discriminated equally difficult intensity changes in the same VCV regions. Attention-related negative difference waves showed enhanced early and late components (Nde and Ndl) during phoneme discrimination conditions. Hemispheric asymmetries developed only during the Ndl and were more pronounced during phoneme discrimination. The results suggest that auditory areas of both hemispheres are specialized for phonetic analysis, with hemispherically specialized mechanisms engaged primarily during the final stages of phoneme processing. PMID- 10380990 TI - Decreased calmodulin-NR1 co-assembly as a mechanism for focal epilepsy in cortical dysplasia. AB - The NMDA receptor is one of the ionotropic glutamate receptors essential for excitatory neurotransmission. The NMDAR1 subunit is inactivated by direct interaction with calmodulin. The protein levels of calmodulin, NMDAR1 and their complex were quantified in tissue resected from epileptogenic and non epileptogenic cortical areas as determined by chronic subdural electrode recordings from three patients (aged 6, 14 and 18 years) with focal epilepsy associated with cortical dysplasia. In all patients, the co-assembly of calmodulin and NMDAR1 was decreased in epileptogenic dysplastic cortex compared with normal appearing non-epileptogenic cortex, while there was no significant difference in the total protein levels of calmodulin or NMDAR1 between the two EEG groups. These results suggest that decreased calmodulin-NMDAR1 co-assembly is a cellular mechanism that contributes to hyperexcitability in dysplastic cortical neurons and in focal seizure onsets. PMID- 10380991 TI - GAT-1 developmental expression in the rat cerebellar cortex: basket and pinceau formation. AB - In view of the key role exerted by neurotransmitter transporters in the synaptic transmission, the expression of GABA transporter GAT-1 was analysed during cerebellar development, when relevant processes of synapse maturation take place. GAT-1-immunoreactive (IR) structures started to be detected on PD 8-9, at the low molecular and Purkinje cell layer, coincident with the onset of functional inhibitory synapses on Purkinje neurons. By PD 18, GAT-1-IR structures completely ensheathed the Purkinje cell somata thus outlining the characteristic perisomatic formation, whereas GAT-1 wrapping on the axon initial segment started to be detected only at PD 15, and the mature form of the pinceau was fully developed from PD 23 on. These results, when compared with the functional maturation of the GABAergic input to Purkinje cells, indicate that GAT-1 may play a significant role in the differentiation of basket interneuron-Purkinje cell circuit. PMID- 10380992 TI - Zinc alleviates thermal hyperalgesia due to partial nerve injury. AB - Zinc has recently been shown to alleviate inflammatory hyperalgesia. In the present study, we showed that intrathecal, intraplantar or systemic injection of zinc chloride significantly relieved thermal hyperalgesia in rats with sciatic nerve injury. Alleviation of thermal hyperalgesia was dose dependent in each case, although higher doses were required for i.p. injections (ED50 = 13.6 nmole) than for intrathecal (ED50 = 0.05 nmole) or intraplantar injections (ED50 = 0.3 nmole). Neither intrathecal nor intraplantar zinc chloride influenced thermal nociception in normal rats without nerve injury. The results provide the first evidence that zinc alleviates neuropathic hyperalgesia. PMID- 10380993 TI - Feeding and general activity patterns of a howler monkey (Alouatta palliata) troop living in a forest fragment at Los Tuxtlas, Mexico. AB - The feeding behavior and general activity patterns of a howler monkey troop living in a 3.6 ha forest fragment were studied at Los Tuxtlas, Mexico, for an annual cycle. Monthly samples of their feeding behavior indicated that they used 52 species of 24 plant families as sources of food. Of these, 67% were trees, which accounted for 96% of total feeding time recorded. Ten species of Moraceae, Cecropiaceae, Anacrdiaceae. and Sapotaceae contributed to 70% of the trees used and to almost 90% of feeding time. The number of plant species used per monthly record varied from 7 to 31 with an average of 19.9 species. Young leaves and ripe fruit were the principal items in the monthly diet of howlers and average percent of time spent consuming these plant parts was 46.7% and 34.8%, respectively. The use of tree species was found to be associated to their importance value and to their pattern of spatial dispersal in the study site. Availability of young leaves was fairly constant from month to month, but it presented a seasonal pattern, and there was a significantly lower number of tree species bearing ripe fruit through the year with brief pulses of production. The monthly activity pattern was found to be related to variations in the availability of young leaves and ripe fruit as well as to the values of the intermonthly overlap in plant species used. Resting and feeding presented a bimodal pattern of occurrence throughout the day that seemed to be related to variations in maximum ambient temperatures. Results are discussed in light of the small size and shape of the forest fragment inhabited by the howler troop. PMID- 10380994 TI - Cranial and mandibular morphometry in Leontopithecus Lesson, 1840 (Callitrichidae, primates). AB - In this paper, we report on a craniometric analysis comparing the species of lion tamarins, Leontopithecus Lesson, 1840. Seventeen cranial and mandibular measures were taken on skulls of 59 adult crania: 20 L. rosalia (14 females and 6 males); 13 L. chrysomelas (6 females and 7 males); 23 L. chrysopygus (8 females and 15 males), and 3 L. caissara (1 female and 2 males). All specimens were from the Rio de Janeiro Primate Center (CPRJ-FEEMA, Brazil), except the specimens of L. caissara. Statistical treatment involved a one-way analysis of variance (the Bonferroni test) and discriminant analysis, comparing cranium and mandibles separately to determine variables which best distinguished groups and to group the specimens, using size corrected methods. The Mahalanobis distance was computed from the centroids of each group. Seven measures distinguished females of L. chrysopygus with L. rosalia, six to L. rosalia with L. chrysomelas, and L. chrysopygus with L. chrysomelas. In males, the numbers of measures statistically different were 5, 4, and 3 of the pairwise comparisons above mentioned. Cranial base length and orbital breadth were the only measures that were significantly different in all three dyads, considering both sexes. For the cranium, function 1 of the Discriminant Analysis accounted for 52.4% of the variance and function 2 accounted for 40.3%. Both functions exhibited a significant value for Wilks' lambda (P<0.0001) and 96.6% of specimens were correctly classified. For the mandible, the first two functions provided a significant discrimination 51.1% and 44.9%, respectively, and 69.5% of the correct classification. Orbital breadth and cranial base length contributed most in the cranial analysis, while mandibular length and mandibular body height to mandibular ones. The analyses performed in this study (univariate and multivariate) demonstrated that cranial and mandibular morphology is significantly different among species of Leontopithecus. Despite of sample size, L. caissara shows morphological distances to L. chrysopygus in cranial analysis. However, other investigations are necessary to confirm this. PMID- 10380995 TI - Trait-like immunological and hematological measures in female rhesus across varied environmental conditions. AB - In this 2-year longitudinal study, 45 2-year-old female rhesus were observed as they were captured and removed from a free-ranging setting (Phase I), single caged for 1 year (Phase II), and housed in small, stable social groups for an additional year (Phase III). During the study, eight blood samples were taken, and hematological, immunological, and hormonal variables were assayed to determine whether 1) any of the measures would exhibit trait-like, inter individual longitudinal stability, despite fluctuations in population means induced by Phases I, II, and III; 2) plasma concentrations of cortisol, prolactin, and norepinephrine would be lowest in Phase III, and elevated during the periods of acute and chronic stress associated with Phases I and II; and 3) there would be any evidence of immunosuppression associated with Phases I and II. The results suggest that the majority of hematological/immunological variables were trait-like throughout the study in contrast to plasma cortisol, prolactin, and norepinephrine concentrations. Thus, red blood cells, hemoglobin, hematocrit, platelets, mean corpuscular volume and hemoglobin, as well as white blood cells, the absolute number of CD4+ (T-helper/inducer) cells, the absolute number of CD8+ (T-suppressor/cytotoxic) cells, total T cells (CD2+%), total B cells (CD20+%), and the ratio of CD4+/CD8+ cells were trait-like. The hematological measures were changed dramatically by capture and the subsequent single caging, with most not recovering to presumed baselines until after 12-28 weeks. The immune measures were depressed at capture (excepting B cells), and during 7 months of single caging failed to return to normal levels associated with social housing. We thus conclude that single housing can produce significant, long-term features of immunosuppression. Capture produced significant increases in plasma cortisol, prolactin, and norepinephrine concentrations. Long periods of single caging produced significant increases in plasma prolactin concentrations, indicative of stress-induced anxiety, and may also have been associated with down-regulation of plasma norepinephrine and cortisol concentrations. PMID- 10380996 TI - Phylogenetic relationships of the Callitrichinae (Platyrrhini, primates) based on beta2-microglobulin DNA sequences. AB - The phylogenetic relationships of callitrichine primates have been determined by DNA sequence analyses of exons 1, 2, and 3 of the beta2-microglobulin gene. Parsimony, distance, and maximum likelihood analyses of ca. 900 base pairs of 21 taxa, representing all callitrichine genera, indicated that Saguinus was the most basal offshoot. Within Saguinus, S. fuscicollis appeared as the first divergent lineage followed by an unresolved trichotomy formed by S. mystax/S. imperator, S. midas/S. bicolor, and S. oedipus. A second callitrichine lineage was formed by Leontopithecus; each of the three species studied showed identical nucleotide sequences. Callimico appeared as the sister taxon of Callithrix/Cebuella. Genetic distances within this latter group were very small, although a stronger association between Cebuella and species of the Callithrix argentata group was observed. The inclusion of Cebuella in the genus Callithrix is suggested. These studies indicated that tamarins are more plesiomorphic than marmosets in agreement with the phyletic dwarfism hypothesis. PMID- 10380997 TI - Intake, digestibility, and passage of a commercially designed diet by two Propithecus species. AB - The digestibility and passage of an experimental diet was used to compare the digestive physiology of two Propithecus species: P. verreauxi and P. tattersalli. Though both animals have a similar feeding ecology, the captive status of P. verreauxi is considered more stable than that of P. tattersalli. The test diet included a local tree species, Rhus copallina, at 15% of dry matter intake (DMI) and Mazuri Leafeater Primate Diet at 85% of DMI. The chemical composition of the diet (dry matter basis) was 25% crude protein, 34% neutral detergent fiber (NDF), and 22% acid detergent fiber (ADF) with a gross energy of 4.52 kcal/g. After a 6 week acclimation to the experimental diet, animals were placed in research caging. After a 7 day adjustment period, animals were dosed with chromium mordant and Co-EDTA as markers for digesta passage and all feed refusals and feces were collected at timed intervals for 7 days. Digestibility values, similar for both species, were approximately 65% for dry matter, crude protein, and energy, and 40% and 35% respectively, for NDF and ADF. Transit times (17-18.5 hr) and mean retention times (31-34 hr) were not significantly different between species, and there was no difference between the chromium mordant and Co-EDTA. Serum values for glucose, urea, and non-esterified fatty acids (NEFA) were obtained during four different time periods to monitor nutritional status. While there was no change in serum glucose, serum urea increased over time. The NEFAs increased across all four time periods for P. verreauxi and increased for the first three periods then decreased in the last period for P. tattersalli. Results obtained indicate no difference in digestibility nor digesta passage between species, and that both Propithecus species were similar to other post-gastric folivores. PMID- 10380998 TI - The full expression of locomotor and motor hyperactivities induced by pressure requires both striatal dopaminergic and N-methyl-D-aspartate receptor activities in the rat. AB - High pressure induced locomotor and motor hyperactivities (LMA), tremor and myoclonia in rat. The LMA has been reported to be reduced by intracerebroventricular (i.c.v.) administration of dopaminergic receptor antagonists. Moreover, the LMA but not myoclonia correlate with pressure induced striatal dopamine increase. Nevertheless the role of dopaminergic and NMDA receptor activities at striatal level in the development of LMA remained unclear. In this study, the microdialysis technique associated to a behavioural device was used to test the effects of intra-striatal administration of D1 antagonist SCH23390 (1 microM), D2 antagonist sulpiride (1 microM) and NMDA antagonist AP-5 (10 microM) on LMA, tremor and myoclonia expression. Data clearly showed that LMA was drastically reduced by each treatment. In contrast, tremor and myoclonia were poorly affected. These data suggest that both dopaminergic and NMDA receptor activities at striatal level are needed for the full expression of the pressure induced LMA and confirm that striatal neurotransmission changes are principally involved in this behavioural disorders. At the light of recent studies on dopaminergic neurotransmission and glutamate evoked-NMDA activity, we suggest that blockage of D1 or D2 receptors should reduced the LMA by reducing glutamate evoked activity. PMID- 10380999 TI - Apoptosis induced by an endogenous neurotoxin, N-methyl(R)salsolinol, is mediated by activation of caspase 3. AB - An endogenous neurotoxin, N-methyl(R)salsolinol, has been proved to be involved in the pathogenesis of Parkinson's disease. Increased level of N methyl(R)salsolinol in the cerebrospinal fluid and high activity of its synthesizing (R)salsolinol N-methyltransferase in lymphocytes were confirmed in the majority of parkinsonian patients. Recently this neurotoxin was found to induce apoptosis in human dopaminergic neuroblastoma SH-SY5Y cells. In this study, we tried to elucidate the intracellular mechanism of apoptosis induced by N-methyl(R)salsolinol, and proved activation of caspase 3 after incubation with this toxin by Western blot analysis. Further, a caspase 3 inhibitor, acetyl-L aspartyl-L-glutamyl-L-valyl-L-aspartic aldehyde, prevented the nucleosomal DNA fragmentation completely. These results demonstrate that caspase 3 mediates apoptosis induced by an endogenous neurotoxin, N-methyl(R)salsolinol, which may cause apoptotic cell death of dopamine neurons in Parkinson's disease. PMID- 10381000 TI - Tryptophan hydroxylase mRNA levels are elevated by repeated immobilization stress in rat raphe nuclei but not in pineal gland. AB - Repeated stress triggers a wide range of adaptive changes in the central nervous system including the elevation of serotonin (5-HT) metabolism and an increased susceptibility to affective disorders. To begin to examine whether these changes are mediated by alterations in gene expression for tryptophan hydroxylase (TPH), the rate-limiting enzyme in 5-HT biosynthesis, we quantitated its mRNA levels by competitive reverse transcription-polymerase chain reaction (RT-PCR). Repeated immobilization stress (2 h, 7 days) elicited a six- or ten-fold rise in TPH mRNA in median raphe nucleus (MRN) and dorsal raphe nucleus (DRN), respectively, without significantly altering TPH mRNA levels in the pineal gland. In contrast, there was little change in mRNA levels for GTP cyclohydrolase I (GTPCH), the rate limiting enzyme in synthesis of the tetrahydrobiopterin (BH4), the obligate cofactor for TPH. This is the first study to reveal stress-elicited activation of TPH gene expression. PMID- 10381001 TI - Decrement of the response of a serotonergic modulatory neuron (the metacerebral cell) in Aplysia, during repeated presentation of appetitive (food) stimuli. AB - Application of food (seaweed, SW) stimuli to the lips evokes a burst of metacerebral cell (MCC) spikes, and it was found in free-moving animals that repeated presentation of the stimulus was associated with a rapid decrement of the evoked responses, even in the absence of ingestion of the food. To aid in discriminating between mechanisms that may be responsible for this decrement, SW was applied repeatedly to the lip ipsilateral or contralateral to one of the paired MCCs, and then generalization of the response decrement was tested by applying a SW stimulus to the opposite (non-stimulated) receptive field. There was statistically significant generalization of response decrement and the amount of generalization appeared to be a function of whether the decrementing stimuli were presented on the side ipsilateral vs. contralateral to the recorded MCC. The overall data suggest that MCC response decrement to repeated food stimuli results in a process analogous to behavioral habituation, and the data are consistent with a simple neural model. PMID- 10381002 TI - Increased NR1-NR2A/B coassembly as a mechanism for rat chronic hippocampal epilepsy. AB - The N-methyl-D-aspartate receptors (NMDAR) produce physiologically functional channels for enhanced excitatory neurotransmission when they exist as heteromeric complexes containing the NMDAR1 subunit combined with NMDAR2. We examined the expressions of NMDAR1 and 2A/B protein in the kainic acid induced rat chronic epileptic hippocampus. Immunoreactivities of both NMDAR1 and NDMAR2A/B were increased in the inner molecular layer of the dentate gyrus, while they were decreased in the hilar and CA3/4 pyramidal zones. Immunoblot analysis demonstrated that the overall level of NMDAR1-2A/B coassembly was increased in the whole hippocampus. These results indicate that the increase of the NMDAR1 2A/B complex in the inner molecular layer is a significant cellular mechanism that contributes to focal hyperexcitability in rat chronic hippocampal epilepsy. PMID- 10381003 TI - Cyclosporine-A reduces spontaneous place preference in adult rats. AB - Cyclosporine-A (CsA) and its analogues have been shown to directly alter locomotor activity. The present study examined CsA effects on spontaneous preferential behavior in adult rats, using a two-shuttle compartment box. The initial preference (>450 s staying time) of the animal for one compartment was measured at pre-conditioning session (900 s). During conditioning session (one 60 min session per day for 6 consecutive days), the animal was alternately injected with CsA (5, 10, 20 and 30 mg/kg per day, i.p.) and vehicle, and its movement restricted to the preferred compartment and the other compartment, respectively. At post-conditioning session (900 s), animals were allowed to freely explore the box. Animals that were treated with 10 mg/kg CsA significantly spent less staying time in the preferred compartment, while those that received other CsA doses displayed a trend of decreased staying time in the preferred compartment. The present data demonstrate that CsA antagonized the spontaneous preferential behavior of animals, and warrant investigations on the drug's utility in altering other preferential behaviors (e.g. drug addiction, alcohol abuse). PMID- 10381004 TI - Roles of nitric oxide in the spinal cord in cardiovascular regulation in rats. AB - The roles of nitric oxide (NO) in the spinal cord in regulation of blood pressure were examined in anesthetized rats. Intrathecal (i.t.) injection of an inhibitor of NO synthase (NOS), N(omega)-nitro-L-arginine methylester (L-NAME), caused marked dose-dependent increase in the blood pressure. The pressor response to L NAME was attenuated by pretreatment with L-arginine (10 micromol, i.t.). Pretreatment with the ganglionic blocker pentolinium (10 mg/kg, i.v.) and alpha1- and beta-adrenoceptor antagonists significantly inhibited the pressor response induced by L-NAME. The pressor responses to L-NAME was blocked by pretreatment with the selective N-methyl-D-aspartate (NMDA) receptor antagonist DL-2-amino-5 phosphonovaleric acid (AP-5) (50 nmol, i.t.), but not by pretreatment with a non NMDA receptor antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). In addition, the pressor response to L-NAME was reduced by spinal cord transection. These results suggest that L-NAME causes activation of the sympathetic nervous system by reduction of NO in the spinal cord, and that this reduction of NO by L NAME may activate glutamatergic neurons in the medulla oblongata. PMID- 10381005 TI - Staurosporine enhances cAMP-induced expression of neural-specific gene VGF and tyrosine hydroxylase. AB - Cyclic AMP transiently stimulates transcription of specific genes in the nervous system, including neural-specific gene VGF. The simultaneous presence of staurosporine (SSP) caused a progressive, rather than transient, increase in VGF mRNA levels during the cAMP-induced differentiation of human neuroblastoma cells. This steady increase does not appear to be due to increased stability of VGF mRNA. This synergy between cAMP and SSP was also observed at the protein level for tyrosine hydroxylase. These findings suggest the presence of cellular machinery that counteracts the decline of cAMP-induced gene expression. PMID- 10381006 TI - Muscle sympathetic nerve response to vestibular stimulation by sinusoidal linear acceleration in humans. AB - To clarify the effects of natural otolith stimulation on muscle sympathetic nerve activity (MSNA) in humans, eight male volunteers were seated in a linear accelerator (sled) during the recording of MSNA from the tibial nerve with microneurography, and also the recording of electrocardiogram, blood pressure measured with a Finapres device and thoracic impedance during movement. Sinusoidal linear acceleration with peak values of +/-0.10, 0.15 and 0.20 Gx were applied to the sitting subjects in the anteroposterior direction. Both the total activity and the burst rate of MSNA decreased during the sinusoidal linear acceleration, whereas the average heart rate, thoracic impedance and mean arterial pressure did not change significantly. These results suggest that moderate sinusoidal linear acceleration in the anteroposterior direction may suppress MSNA in humans. PMID- 10381007 TI - Effect of 0.2 T static magnetic field on human neurons: remodeling and inhibition of signal transduction without genome instability. AB - We describe the effect of the static magnetic field generated by a 0.2 T magnetic resonance tomograph on a normal human neuronal cell culture (FNC-B4). After 15 min exposure cells showed dramatic changes of morphology: they formed vortexes of cells and exposed branched neurites featuring synaptic buttons. At the same time, thymidine incorporation and inositol lipid signaling were significantly reduced. Control (sham exposed) or non-neuronal cells (mouse leukemia, and human breast carcinoma cells) did not show any alteration following exposure. Endothelin-1 release from FNC-B4 cells was also dramatically reduced after 5 min exposure. However, PCR analysis of 12 DNA microsatellites selected as gauges of genome instability, did not reveal any alteration following exposure, thus ruling out a direct effect of the magnetic field on DNA stability. These data can be interpreted as a specific effect of the static magnetic field on human neuronal cells and are consistent with the induction of remodeling and differentiation; they demonstrate that fields below 0.5 T have significant biological effects on human neurons. PMID- 10381008 TI - Visual contribution to self-induced body sway frequencies and visual perception of male professional dancers. AB - We studied the degree of dependence on vision, for postural control and for perception, among male adult dancers and untrained subjects. First, body sways were analyzed on a free seesaw platform. Fast Fourier transform processing allowed spectral frequency analysis of the platform sways recorded by an accelerometer. Secondly, a visual dependence test, the rod and frame test (RFT) was used. Professional dancers were significantly more stable and less dependent on vision for postural control and for perception than untrained subjects. Presumably, professional dance training strengthens the accuracy of proprioceptive inputs and shifts sensorimotor dominance from vision to proprioception. For the dancers, there was interaction between the RFT visual dependence and the visual control of posture: the less visual-dependent they were for the RFT, the more stable they were in dynamic balance conditions. PMID- 10381009 TI - Immunocytochemical localization of nitric oxide synthase in the mammalian retina. AB - The localization of nitric oxide synthase (NOS) was investigated by immunocytochemistry and immunoblotting using an antiserum against neuronal NOS in the rat, mouse, guinea pig, rabbit and cat retinae. Western blot analysis of retinal tissue extracts showed that the NOS-immunoreactive band of 155 kDa was present in all species. In the rat, mouse, guinea pig and rabbit retinae, two types of amacrine cells and a class of displaced amacrine cells were consistently NOS-labeled. In the cat retina, unlike other mammals, one type of amacrine cells and two types of displaced amacrine cells showed NOS immunoreactivity. NOS immunoreactivity was further found in some bipolar cells of the rat and guinea pig, some interplexiform cells of the mouse, some photoreceptor cells of the rabbit and some Muller cells of the cat. PMID- 10381010 TI - Preparatory balance adjustments precede withdrawal response to noxious stimulation in standing humans. AB - Self-initiated leg movement in standing humans is preceded by a medio-lateral preparatory balance adjustment (PBA); however, such preparatory balance control is often absent in reflex-like stepping responses evoked by whole-body instability. The presence or absence of the PBA may reflect a task-dependent modulation of the response serving to preserve lateral stability (PBA present) or avoid delay in the lifting of the foot (PBA absent). To examine whether such task dependent modulation can occur during more stereotypical limb movements, we examined spinally-mediated withdrawal responses evoked by noxious stimulation of the foot. Results showed that rapid limb withdrawal was preceded by a large PBA when subjects were standing but not when they were supine. The PBA caused limb withdrawal to the noxious stimulation to be delayed. However, the onset of the PBA in the standing trials was equivalent in timing to the onset latency of the classic withdrawal responses recorded during the supine trials. Evidence of a preparatory balance adjustment evoked, in advance of a delayed withdrawal response, at very rapid latencies (underlying muscle activation at 70-120 ms) may raise new questions about the neural mechanisms underlying the co-ordination of balance and movement. PMID- 10381011 TI - Release of mitochondrial cytochrome c and DNA fragmentation after cold injury induced brain trauma in mice: possible role in neuronal apoptosis. AB - Recent studies have shown that release of mitochondrial cytochrome c is a critical step in the apoptosis process. In this study, we examined the subcellular distribution of the cytochrome c protein after cold injury (CI), in which apoptosis is assumed to participate. Western blotting and immunohistochemistry showed cytosolic cytochrome c as early as 1 h after CI, and correspondingly, there was a reduction in mitochondrial cytochrome c after injury. Neuronal distribution of cytosolic cytochrome c was shown by double staining with a neuronal nuclear marker by immunohistochemistry. A significant amount of DNA laddering was detected 4 h after CI, and increased in a time dependent manner. These data suggest that early cytochrome c release from mitochondria may contribute to apoptosis induction after traumatic brain injury. PMID- 10381012 TI - Maternal nicotine depresses eupneic ventilation of neonatal rats. AB - Maternal smoking is a risk factor for the sudden infant death syndrome (SIDS). We hypothesized that pre-natal exposure to nicotine would result in abnormalities of ventilatory activity in newborns. Neonatal rats which had been exposed to nicotine had significantly lower minute ventilation breathing air and hypoxic gas mixtures than did control animals. In both groups, anoxia elicited gasping which was equally effective in restoring eupnea. Maternal exposure to nicotine may result in a reduced metabolic rate and/or chronic hypoventilation in the newborn. PMID- 10381013 TI - North of England evidence-based guideline development project: summary version of guidelines for the choice of antidepressants for depression in primary care. North of England Anti-depressant Guideline Development Group. PMID- 10381014 TI - How often is the diagnosis bronchial asthma correct? AB - BACKGROUND: There are studies indicating that bronchial asthma is often underdiagnosed, while only a little research has been conducted as concerns overdiagnosing asthma. OBJECTIVE: We aimed to estimate the number of patients who have been given the wrong diagnosis of asthma. METHODS: All patients aged above 18 years who had visited two GPs during 1994 or 1995, with the diagnosis of bronchial asthma confirmed in the medical register, were examined by a specialist in allergies. RESULTS: One hundred and twenty-three patients fulfilled the criteria for being included in the study. Eighty-six patients (70%) attended the examination. Of these, 51 (59%) had bronchial asthma, six (7%) asthma in combination with chronic obstructive pulmonary disease (COPD) and 29 (34%) no asthmatic disease. CONCLUSION: The study indicates that more accuracy is needed when diagnosing bronchial asthma. PMID- 10381015 TI - Self-treatment of asthma: possibilities and perspectives from the practitioner's point of view. AB - OBJECTIVES: Self-management of asthma is becoming more and more widespread. The implementation of this treatment strategy requires changes in the role and attitude of the GP. These changes may be hindered by obstacles both expected and experienced. As self-treatment of asthma is more common in the UK, comparison between UK and Dutch GPs provides a good opportunity to identify possible obstacles in general practice to the implementation of self-treatment of asthma with inhaled corticosteroids. METHODS: We carried out a qualitative descriptive study with self-administered questionnaires and interviews. Questionnaires were sent to 500 randomly selected Dutch GPs. Interviews were held with 20 Dutch and 25 British GPs in order to acquire more in-depth information. The outcome measures were attitude towards, knowledge regarding and experiences with self treatment of asthma; organizational requirements; and expectations of consequences of self-treatment in general practice. RESULTS: The Dutch and British GPs investigated have a positive attitude towards self-treatment of asthma. Though knowledge about self-treatment is present among a majority of the GPs, self-treatment by patients is not yet as common in The Netherlands as it is in the UK. Nineteen per cent of the Dutch GPs had experience with a written peak flow-based self-treatment plan related to the usage of inhaled steroids. According to our findings, present expected obstacles are probably mainly of the organizational kind, such as the availability of time, money and materials. CONCLUSIONS: There is a positive attitude towards the implementation of self treatment plans in general practice, but problems relating to certain identified obstacles need to be addressed. There is a need to define which patients might profit from self-treatment, and further proof of both the clinical effectiveness and the cost-effectiveness of self-treatment needs to be acquired. PMID- 10381016 TI - Better by half: hypertension in the elderly and the 'rule of halves': a primary care audit of the clinical computer record as a springboard to improving care. AB - BACKGROUND: Despite recent studies highlighting the benefits of treating elderly hypertensives, researchers have shown that the taking on board of these findings has been disappointing in primary care, where the 'rule of halves' still applies. Clinical computers could help performance in this area, yet national and local research suggests that they are under-used. OBJECTIVE: Our aim is to develop a pragmatic intervention which aims to: improve patient care by translating research findings into practice, increase meaningful computer use, establish 'paperless' annual audits and improve 'networking' between practices. METHOD: Following a baseline audit to ascertain accuracy, the computer records of participating practices were tested against the 'rule of halves' for hypertension. Results were presented to each practice (individual practice and aggregate data for all practices). Management guidelines, standardization of computer recording, achievable targets and review dates were agreed. The study was conducted in West London practices using the EMIS computer system in 1996/1997. RESULTS: An 81% (22/27) practice response rate was achieved. Baseline audit was completed for 22 practices. Fifteen practices appear to be using their computer regularly (two-thirds). Using strict definitions, 'the rule of halves' still applies. Using looser definitions, three-quarters of hypertensives are known, two-thirds are treated and just under two-thirds are controlled. This project identified wide inter- and intra-practice variation in: use of the computer, patient follow-up, attainment of target BP, rounding BP readings to target levels and prescribing patterns. CONCLUSION: This focused training intervention has introduced practices to evidence-based proactive care and highlighted an important application for clinical computers. A local network of practices has been established for future projects. For elderly patients registered with a GP, the rule of halves has been improved upon, provided that a figure of 160/90 is taken as an adequate control. Attainment of target BP in treated hypertensives was similar to that reported from large trials. There is enormous scope for improving identification and follow-up of hypertensives using clinical computers and systematic models of care. The wide inter-practice variation in hypertension management requires further study. PMID- 10381017 TI - The prognostic value of gastrointestinal morbidity for gastric cancer. AB - OBJECTIVE: A history of gastrointestinal disease is a risk factor for organic gastrointestinal disorders overall. We conducted a retrospective case-control study to explore whether a history of gastrointestinal disease is of prognostic value for gastric cancer. METHODS: Forty-six patients with gastric cancer were identified from a dynamic population of approximately 12,000 patients followed since 1971 in four general practices. The control subjects without gastric cancer were matched one-on-one according to age, general practice, sex and observation period. Data on gastrointestinal morbidity in the period before gastric cancer was diagnosed were obtained from the Continuous Morbidity Registration. RESULTS: The mean observation period between the date of enrolment in the registration and the first diagnosis of gastric carcinoma was 12 years. Although every patient with gastric cancer ultimately will develop gastrointestinal complaints, 28 of these patients had no previous gastrointestinal morbidity, in comparison with 26 control subjects. Furthermore, patients who developed gastric cancer did not have more frequent gastrointestinal morbidity in their past than the control subjects (odds ratio 0.80, 95% CI 0.32-2.03). CONCLUSIONS: Our results suggest that a history of gastrointestinal morbidity is not of prognostic value for gastric cancer. Focusing attention on patients with a past history of gastrointestinal symptoms to detect gastric cancer may be of little value. PMID- 10381018 TI - The impact of type 2 diabetes mellitus on daily functioning. AB - BACKGROUND: Traditionally, health and the outcomes of medical treatment have been measured in terms of morbidity, incidence or prevalence of disease, or even mortality. This disease model provides an adequate framework for acute illnesses, but for chronic diseases, severity and their effect on everyday functioning are paramount. For chronic diseases, functional health status, as a vital part of quality of life, is now recognized as an important outcome measure of the GP's care. OBJECTIVE: We aimed to assess the impact of type 2 diabetes mellitus on functional health status in Dutch general practice. METHOD: We conducted a cross sectional study of the functional health status of all patients with type 2 diabetes mellitus under 85 in two general practices, using the Sickness Impact Profile (SIP) and the COOP/WONCA charts. A control group of non-diabetic patients was selected, matched for practice, sex and age. RESULTS: In total, 127 type 2 diabetes mellitus patients and 127 controls participated in the study, the responses being 78 and 70%, respectively. Between these groups the following were significantly different: the SIP subscore Physical, the SIP sum score and the COOP/WONCA scores for physical fitness and overall health. Type 2 diabetes mellitus patients were 2.46 (95% CI 1.5-4.1) times more likely to experience functional impairment. Cardiovascular morbidity (odds ratio 2.5, 95% CI 1.3-4.7), locomotory morbidity (odds ratio 2.6, 95% CI 1.4-5.1) and diabetes itself (odds ratio 1.4, 95% CI 1.1-1.9) were significantly associated with the presence of functional impairment. CONCLUSION: This study demonstrates the impact of type 2 diabetes mellitus on functional status, particularly in relation to cardiovascular morbidity. PMID- 10381019 TI - Referral for 'prostatism': developing a 'performance indicator' for the threshold between primary and secondary care? AB - OBJECTIVE: We aimed to define a performance indicator at the gateway between primary and secondary care. METHOD: We carried out an analysis of referral letters sent to an urological department within the catchment area of a teaching hospital in Cardiff, Wales. The subjects were 221 sequential referral letters from 221 GPs. The main outcome measures were the information content of referral letters analysed. Letters were stratified into referral threshold groups by the presence of history, examination, routine investigations and specialized investigations. RESULTS: Three distinct categories of referral practice were identified: referrals which contained history alone; those providing history examination and a selection of routine investigations; and those providing history, examination data and the results of routine and specialized investigations. The study demonstrated that more than a third of GPs do not report the results of digital rectal examination in their referrals and only 4% record urinary flow rates and post-micturition residual urine volume. CONCLUSIONS: The majority (60%) of generalist referrals to an urology department for prostatism provide enough information for specialists to be able to prioritize appointments, but more than a third (36%) of the referrals contain inadequate information. The method has the potential of being developed into a gateway performance indicator in clinical practice. PMID- 10381020 TI - Pathways from symptoms to medical care: a descriptive study of symptom development and obstacles to early diagnosis in brain tumour patients. AB - BACKGROUND: The time between experiencing symptoms and treatment in cancer diseases is a time of insecurity and despair. Brain tumour disease is a severe disease with dramatic manifestations and it is important that this time be kept as short as possible. METHODS: A consecutive sample of 28 patients with malignant gliomas and their spouses were interviewed about symptom development, help seeking and experiences of medical care. The cumulative development of their symptoms was described and factors acting as obstacles to medical care were identified. RESULTS: Most spouses witnessed months of global dysfunction preceding the symptom leading to physician consultation. The patient factors 'less alien symptoms', 'personality change' and 'avoidance'; the spouse factors 'spouse's passivity' and 'spouse's successive adaptation'; and the physician factors 'reasonable alternative diagnosis', 'physician's inflexibility' and 'physician's personal values' were identified as obstacles on the pathway to appropriate medical care. The importance of acknowledging the power of the spouse as a provider of substantial information from everyday life facilitating differential diagnosis is stressed. PMID- 10381021 TI - Outreach clinics in Israel: a common but unregulated phenomenon. AB - BACKGROUND: Specialist outreach consultations in the primary care setting have long been controversial with regard to both their effectiveness in treating patients and their potential in improving the interaction between family physicians and specialists. OBJECTIVE: The aim of this study was to establish the prevalence and nature of outreach consultations in primary care clinics in Israel. METHODS: Questionnaires were sent to the heads of all public family practices of the General Sick Fund in our district (38 practices with about 180,000 patients). All 38 practice managers returned completed questionnaires. RESULTS: Twenty-eight of the 38 practices (74%) have some type of specialist consultation available within their clinics. The most common specialties providing outreach clinics are cardiology (47%), nephrology (45%) and internal medicine (39%), where the consultation was performed with the family physician and the patient present. Psychiatry consultations (42%), however, were generally performed without the patient being present. Most of the practice heads felt that in essence outreach clinics could be a positive way of treating their patients. CONCLUSIONS: Head physicians of primary care clinics tend to see outreach clinics as being a very positive tool with which to treat patients. Although many family physicians have some form of specialist consultation available, it is provided and performed mainly on an ad hoc basis. At present no data are available on how best to structure these consultations, or on which specialties outreach clinics are most suitable. PMID- 10381022 TI - The vagaries of self-reports of physical activity: a problem revisited and addressed in a study of exercise promotion in the over 65s in general practice. AB - BACKGROUND: The assessment of levels of physical activity relies upon suitable measurement tools. OBJECTIVE: We aimed to investigate whether a practice nurse, using a motivational interview technique, could encourage older patients to increase their physical activity. METHODS: Health and well-being were monitored at baseline and 8 weeks following intervention. Physical activity levels were ascertained using both a self-report measure and ambulatory heart-rate monitoring. RESULTS: Whilst patients reported higher levels of physical activity at follow-up, this finding was not confirmed by the heart-rate data. CONCLUSION: The study concludes that patients tend to overestimate the amount of physical activity undertaken and that ambulatory heart-rate monitoring may be more useful for verifying actual behaviour. PMID- 10381023 TI - Attitudes to cardiovascular health promotion among GPs and practice nurses. AB - BACKGROUND: Cardiovascular health promotion is an important element of national health strategy, but doubts have been raised about current methods, and attitudes among general practice staff are ambivalent. OBJECTIVES: We aimed to assess attitudes to cardiovascular health promotion, opinions about efficacy and perceptions of skills in lifestyle counselling in GPs and nurses from the same practices. METHOD: A questionnaire survey of 107 GPs and 58 practice nurses from 19 group practices (100% response rate). RESULTS: Practice nurses were seen to have the main responsibility for cardiovascular health promotion. Although attitudes to health promotion were generally positive, lack of training in lifestyle counselling was perceived to be a problem. Few responders believed that they were very influential in helping people change their lifestyles. Beliefs about the effectiveness of lifestyle counselling were mixed, with cigarette smoking, physical inactivity and obesity being seen as difficult to change. Beliefs in the effectiveness of lifestyle counselling were associated with positive attitudes towards health promotion and greater confidence in training. No association between personal health behaviour and attitudes towards health promotion were observed. CONCLUSIONS: It is recognized that health promotion involves more than the provision of simple information and advice, but GPs and practice nurses lack confidence in lifestyle counselling skills. The attitudes of health professionals are crucial to the implementation of prevention strategies and require regular review. PMID- 10381024 TI - The role of general practice in promoting teenage health: a review of the literature. AB - BACKGROUND AND METHODS: Teenagers are acknowledged to be at high risk of health damaging behaviours including smoking, teenage pregnancy, and drug and alcohol use. Additionally, the recognition of high levels of psychological distress is cause for serious concern about teenage health. This paper reviews health promotion interventions for teenagers in general practice. Medline, BIDS, Psyclit and SIGLE databases for January 1990-February 1997 were systematically searched for English language studies on adolescent/teenage health and health promotion interventions in primary health care/general practice; reference sections of articles were checked for earlier work. CONCLUSIONS: The literature indicates that teenagers rarely receive health promotion advice from their physicians. The impact on behaviour change, of screening and health promotion for teenagers in general practice requires further evaluation to asssess the potential effectiveness in preventing the onset or continuation of health-damaging behaviours. PMID- 10381025 TI - Combining qualitative interviews with video-recorded consultations: gaining insight into GPs' decision-making. AB - BACKGROUND: Studies of GPs' decision-making are important for facilitating our understanding of GPs' consulting behaviours. We have used a novel combination of semi-structured interviews and video-recorded consultations to research the influences on decisions made by GPs during their consultations. OBJECTIVE: We describe the use of GPs' video-recorded consultations as a stimulus for focused, semi-structured interviews and to discuss how this research method compares with other approaches for studying GPs' decision-making during consultations. METHODS: GPs' surgery sessions were video-recorded and later they were shown video recordings of themselves consulting with smokers before participating in semi structured interviews about these consultations. Interviews aimed to describe the factors which GPs perceived to influence their decisions to discuss or not discuss smoking with patients. DISCUSSION: This technique can be used to research decisions, which are made frequently by GPs. It is probably most appropriate for gaining insight into decision-making during mundane consultations, to which GPs would otherwise give little thought. PMID- 10381026 TI - Computer-assisted telephone interview (CATI) in primary care. AB - OBJECTIVES: We aimed to study the prevalence of cardiovascular disease (CVD) risk factors among 11,000 inhabitants in Northern Helsinki, and to identify high-risk individuals in the area and direct them to the local primary-health-care-centred CVD-risk-factor prevention programme. METHOD: We conducted a computer-assisted telephone interview (CATI), a descriptive survey and primary care unit searching for CVD risk factors within the population under its responsibility. Six hundred and sixty-seven individuals aged 18-65 years out of 1000 randomly chosen inhabitants were interviewed using CATI. We measured the prevalence of self reported CVD risk factors: smoking, blood pressure, last measured total serum cholesterol, body mass index (BMI), alcohol consumption, diabetes, physical exercise habits, positive family history of CVD/diabetes and personal history of CVD. RESULTS: Sixty-seven per cent of the sample was interviewed. Nineteen per cent did not have a telephone and 3% refused to be interviewed. Eleven per cent did not respond. Persons with high cardiovascular risk scores were observed mainly in the oldest age group. In the total sample, 23% of women and 28% of men were estimated to be at high risk of coronary artery disease. Gender differences were seen only in one age-group: 45-54-year-old men reporting higher risk-factor scores. The results were analysed using the Statistical Analysis System (SAS). CONCLUSIONS: The CATI-method is a useful tool in screening of high-CVD-risk patients and in guiding them to local CVD primary prevention programmes. PMID- 10381027 TI - How does a change in the administration method affect the reliability of the COOP/WONCA Charts? World Organization of National Colleges, Academies and Academic Associations of General Practitioners/Family Physicians. AB - BACKGROUND: An interviewer is often needed to administer the COOP/WONCA Charts to Chinese patients, and this may affect the reliability of results. OBJECTIVES: We aimed to find out the reliability of the COOP/WONCA Charts administered by an interviewer, and whether a change in the interviewer or administration method would affect the results. METHODS: We carried out a cross-sectional test-retest study on 487 Chinese adult patients attending a family medicine clinic in Hong Kong. The COOP/WONCA Charts were administered by the same interviewer, two different interviewers or self-completion and interviewer administration, on test and retest. The random, inter-observer and inter-method variances were compared with the inter-subject variance. The reliability coefficient of each COOP/WONCA Chart was calculated for each method of administration. RESULTS: Random errors could change the scores by 0.57-1.04, inter-observer variations could change the scores of four charts by 0.72-0.80, and a change in the method could change the physical fitness score by 1.79 and the daily activities score by 1.31, on a five point scale. The reliability coefficients of the six COOP/WONCA Charts were 0.68 0.92 for one interviewer, 0.59-0.82 for two interviewers and 0.46-0.81 for two methods. CONCLUSION: The Chinese COOP/WONCA Charts were reliable in detecting real differences when administered by an interviewer. A change in the method of administration significantly decreased the reliability of the results. The use of more than one method of data collection in the same survey should be discouraged. PMID- 10381028 TI - Questions you didn't ask? COOP/WONCA Charts in clinical work and research. World Organization of Colleges, Academies and Academic Associations of General Practitioners/Family Physicists. AB - OBJECTIVE: COOP/WONCA Functional Assessment Charts are widely in use in research and clinical work. They originally evolved from Dartmouth COOP Functional Assessment Charts. Our objective is to describe our experiences with COOP/WONCA Charts and to provide data on reliability and reference data from a normal population. METHOD: A test-retest study comprising 40 randomly selected patients attending GPs' surgeries was conducted. Eight GPs took part in this study. The response rate for the questionnaire was 100%. In a second study, 245 asthma patents who consulted 53 different GPs were consecutively included; 171 of these repeated completion of the charts after 6 months. Mean scores from a random sample of 2864 persons in the total population of the municipality of Ullensaker in Norway are given as reference values. There was a postal return rate of 64%. RESULTS AND CONCLUSIONS: The reliability of the charts is good, and nine out of ten of the patients gave clinically meaningful answers after 2-3 days. The scores of the 171 asthma patients were nearly identical after 6 months. The reference values are presented. A standardized method for reliability testing is discussed, and the use of COOP/WONCA Functional Assessment Charts is commented on. PMID- 10381029 TI - Selections from current literature: effects of dieting and exercise on resting metabolic rate and implications for weight management. PMID- 10381030 TI - Selections from current literature: smoking and depression. PMID- 10381031 TI - Bone marrow transplant for rheumatoid arthritis: the costs of a cure. PMID- 10381032 TI - Classification criteria for reactive arthritis. PMID- 10381033 TI - Problems and advances in the diagnosis of reactive arthritis. PMID- 10381034 TI - Effects of interleukin 1 receptor antagonist alone and in combination with methotrexate in adjuvant arthritic rats. AB - OBJECTIVE: To determine the benefit of combination treatment with the interleukin 1 receptor antagonist (IL-1ra) and methotrexate (MTX) in adjuvant arthritic rats. METHODS: Rats with adjuvant arthritis were treated by continuous sc infusion with IL-1ra (5 mg/kg/h). Effects of IL-1ra treatment alone were compared to treatment with daily oral MTX (0.048, 0.06, or 0.075 mg/kg) or MTX in combination with IL 1ra. Efficacy was monitored by sequential caliper measurement of ankle joints, final paw weights, and histologic evaluation with particular emphasis on bone lesions. RESULTS: Treatment with IL-1ra alone resulted in 6% inhibition of paw swelling assessed by final paw weight. Treatment with MTX (0.075 mg/kg PO) gave 47% inhibition and the combination resulted in an 84% decrease in swelling. Histologic evaluation of ankle joints revealed 53% inhibition of bone resorption with IL-1ra alone, 58% inhibition with MTX alone, and 97% inhibition in combination treated rats. Lower doses of MTX in combination with IL-1ra also provided additive benefit on arthritis variables. CONCLUSION: Combination therapy with IL-1ra and MTX results in additive or synergistic benefit, with excellent inhibition of all arthritis variables. These data support clinical investigation of the use of combination therapy of IL-1ra and MTX in patients with rheumatoid arthritis. PMID- 10381035 TI - The SHAP-HA complex in sera from patients with rheumatoid arthritis and osteoarthritis. AB - OBJECTIVE: To investigate serum derived hyaluronan (HA) associated protein hyaluronan (SHAP-HA) complex in sera from patients with rheumatoid arthritis (RA) and osteoarthritis (OA) and determine levels of the complex in comparison with those of hyaluronan (HA), in order to assess the role of the complex as an indicator of joint inflammation. METHODS: ELISA and HA binding assays were used for quantitation of the SHAP-HA complex and HA levels in serum samples from 142 patients (114 with RA, 28 with OA) and 31 healthy controls. Clinical evaluations were also performed. RESULTS: In some RA sera, SHAP-HA complex levels were extremely high compared to control levels, but in OA sera no marked increase was observed compared to controls. This was also the case with the HA levels compared between RA and OA sera. However, in RA the levels of the SHAP-HA complex appear to be more related to clinical variables than are levels of HA, and the most significant correlations between levels of SHAP-HA complex and HA were found in the RA group. CONCLUSION: Quantitation of the SHAP-HA complex in sera may be useful as a joint marker that directly correlates to the degree of joint inflammation in RA, and offers new insight into the pathogenesis of arthritis. It may also serve as an independent criterion in the subsequent classification of RA and OA. PMID- 10381036 TI - Incidence of acromioclavicular joint involvement in rheumatoid arthritis: a 15 year endpoint study. AB - OBJECTIVE: To evaluate the incidence of involvement and nature of destruction of acromioclavicular joints (AC) in a prospectively followed cohort of 74 patients with rheumatoid factor positive and erosive rheumatoid arthritis (RA). METHODS: At the 15 year followup, radiographs of 148 AC joints were evaluated, and the grade of destruction was assessed by the Larsen method. RESULTS: No surgical procedures had been performed on the AC joints. Rheumatoid involvement (Larsen Grade > or = 2) was observed in 87/148 (59%) of the AC joints in 50/74 (68%) patients: 37 bilaterally and 13 unilaterally. Incidence of mild erosions (Larsen Grade 2) was 39%, and of severe (Larsen 3-5) 20%. Erosions were most often observed on the inferior edge of the clavicular joint margin. Degenerative features without rheumatoid changes were present in 11 joints. Larsen score (0 100) of peripheral joints correlated well with the AC joint Larsen Grade in both sides: right, r = 0.56 (95% CI 0.38 to 0.70), left, r = 0.49 (95% CI 0.30 to 0.65). CONCLUSION: After 15 years two-thirds of the patients with RA showed involvement of the AC joints. Erosions were located most often on the inferior margin of the joint. PMID- 10381037 TI - Correlation between radiation dose, synovial thickness, and efficacy of radiosynoviorthesis. AB - OBJECTIVE: To correlate the therapeutic efficacy of radiosynoviorthesis (RSO) to radiation doses achieved. METHODS: In 20 patients with known rheumatoid arthritis, radiosynoviorthesis was performed in 36 joints. Arthritis disease activity was assessed by "blood pool scintigraphy" (n = 29) score after injection of 370 MBq 99mTc-MDP, before and at 1, 2, and 5 months after the RSO in 12 patients. For semiquantitative measurements, a region-of-interest technique was applied. Synovial thickness and response to the RSO were evaluated by joint ultrasonography. Pain levels were evaluated semiquantitatively. Dosimetry (n = 20) was calculated using planar quantification according to the MIRD scheme. RESULTS: The mean radiation absorbed dose of 186Re-sulfate to the whole body was 5.3+/-2.7 cGy, liver 10.0+/-8.1 cGy, spleen 20.3+/-22.9 cGy, kidneys 9.4+/-11.4 cGy, and at the injected joints of the shoulder 120.5+/-32.2 Gy, hand 130.0+/ 12.6 Gy, elbow 83.6+/-38.7 Gy, and talar/subtalar joint 84.1+/-30.7 Gy. In 7 cases, where mandatory immobilization of the joint was not possible, the dose to the lymph nodes (n = 25) was 25.9+/-53.8 Gy (maximum 189 Gy) and to single lymph nodes 14.6+/-11.2 Gy (maximum 63 Gy). The reduced doses to the synovia (at 40% leakage) were: 169Er-citrate 73.10+/-25.25 Gy; 90Y-citrate 59.25+/-46.45 Gy; 186Re-sulfate 65.40+/-32.55 Gy. In cases of complete immobilization, the dose to the lymph nodes was negligible: 169Er-citrate (n = 7), whole body dose 0.4 cGy, lymph nodes 2.3 Gy, finger joints 132.3+/-34.3 Gy; 90Y-citrate (n = 6), whole body dose 15.5 cGy, liver dose 26.5 cGy, splenic dose 11.9 cGy, kidney dose 67 cGy, joint knee joint dose 130.1 Gy. Regarding therapeutic effect, mean reduction of the 99mTc-MDP blood pool activity was 41% at first month, 48% at second month, 48% at the fifth month, 48% in larger joints, and 63% in finger joints. Three and 6 months after RSO, sonography showed a mean reduction in synovial swelling: in the knee joint 1.67 and 4.38 mm, respectively; in the larger joints (shoulder, elbow, hand, talar/subtalar) 0.88/1.93 mm; and in finger joints 0.53/1.76 mm. Clinically, best results were observed in the finger joints. CONCLUSION: 1. We observed a significantly higher radiation absorbed dose to the lymph nodes and lower dose to the synovia in the absence of joint immobilization. Immobilization of the joint is essential. 2. At 2 months after treatment, a significant reduction of blood pool activity and synovial swelling was observed, with further improvement in the following months, especially in the finger joints. 3. There is a strong correlation between the reduction of blood pool activity, synovial swelling, and improvement of pain. PMID- 10381038 TI - Serum concentrations of interleukin 6, osteocalcin, intact parathyroid hormone, and markers of bone resorption in patients with rheumatoid arthritis. AB - OBJECTIVE: To analyze serum concentrations of interleukin 6 (IL-6), osteocalcin, intact parathyroid hormone (PTH), and type 1 collagen carboxyterminal telopeptide (ICTP) as well as the urinary concentrations of crosslinked N-telopeptides of type 1 collagen (NTx) and deoxypyridinoline (Dpd) in patients with rheumatoid arthritis (RA) to investigate their role in the etiology of the osteopenia in this disease. METHODS: Using ELISA and radioimmunoassay methods, we estimated serum concentrations of IL-6, osteocalcin, ICTP, intact PTH, and spot urine concentrations of NTx and Dpd in 25 female patients with active RA, 25 female patients with suppressed disease, and 25 age matched healthy female controls. RESULTS: Patients with active RA had significantly higher (p < 0.001) concentrations of IL-6 (94.0+/-12.1 pg/ml) compared to patients with suppressed disease (13.2+/-0.8 pg/ml) and healthy controls (12.3+/-0.8 pg/ml). Serum osteocalcin was significantly lower (p < 0.001) in patients with active RA (1.9+/ 0.2 ng/ml) compared to patients with suppressed disease (2.7+/-0.2 ng/ml) and the controls (2.9+/-0.2 ng/ml). Similarly, serum intact PTH was significantly lower (p < 0.001) in patients with active disease (29.9+/-1.5 ng/ml) compared to patients with suppressed RA (38.0+/-1.6 ng/ml) and controls (49.8+/-2.4 ng/ml). Serum ICTP was also significantly higher (p < 0.01) in patients with active RA (9.5+/-0.3 microg/l) versus patients with suppressed disease (4.1+/-0.2 microg/l) and controls (3.4+/-0.2 microg/l). In patients with active disease, spot urine concentrations of NTx (123.1+/-5.1 nmol bone collagen equivalent/mmol creatinine) and Dpd (15.1+/-0.7 nmol/mmol creatinine) were significantly higher (p < 0.001) than in patients with suppressed disease (58.4+/-2.5 nmol bone collagen equivalent/mmol creatinine and 10.1+/-0.5 nmol/mmol creatinine, respectively) and healthy controls (53.4+/-2.1 nmol bone collagen equivalent/mmol creatinine and 9.7+/-0.5 nmol/mmol creatinine, respectively). There were no significant correlations between serum IL-6 and serum ICTP (r = 0.2357, p = 0.257) or urinary NTx (r = 0.1436, p = 0.494) or between serum intact PTH and ICTP (r = 0.0206, p = 0.922) in patients with active RA. CONCLUSION: There are no significant correlations between bone resorption markers and serum IL-6 and intact PTH in patients with RA. PMID- 10381040 TI - Identification and prevalence of rheumatoid nodules in the finger tendons using high frequency ultrasonography. AB - OBJECTIVE: Rheumatoid nodules are classical diagnostic feature of rheumatoid arthritis (RA). The prevalence of rheumatoid nodules in the finger tendons is not known. We determined the prevalence of rheumatoid nodules of finger tendons in patients with RA, using high frequency linear transducers with high resolution ultrasonography. METHODS: The study comprised 54 consecutive patients with RA and 20 controls. Dynamic ultrasound examination of various tendons was performed using real-time equipment, the Apogee 800 ATL, with an 11 MHz linear array transducer. RESULTS: RA nodules were identified in 9 patients (16.66%); their sizes ranged from 0.21 to 0.95 cm (mean 0.35+/-0.12 cm), in 4 cases (7.4%) the nodules were intratendinous. The flexor tendons were affected in the 9 patients. The finger tendon ultrasonography identified RA nodules that had escaped routine clinical detection. CONCLUSION: The ultrasonography technique was valuable in the confirmation of rheumatoid nodule involvement of the finger tendons. The prevalence of rheumatoid nodule in finger tendons was 16.66%. We believe that ultrasonography should be the screening procedure of choice for diagnosis of RA nodules of the fingers. PMID- 10381039 TI - Dynamic strength training in patients with early rheumatoid arthritis increases muscle strength but not bone mineral density. AB - OBJECTIVE: To assess the effects of 12 months' dynamic strength training on muscle strength and bone mineral density (BMD) at the lumbar spine and femoral neck in patients with early rheumatoid arthritis (RA). METHODS: Thirty-two subjects in the training group (EG) and 33 in the control group (CG) completed the study. EG carried out strength training 2 times a week with moderate loads of 50-70% of repetition maximum. They were also encouraged to do recreational physical activities. CG performed recreational physical activities and range of motion exercises. Maximal strength of the knee extensors, trunk extensors and flexors, and grip strength were recorded with dynamometers. BMD was measured using dual x-ray absorptiometry. Modified Disease Activity Score, erythrocyte sedimentation rate, and pain were used for the estimation of disease activity, and Stanford Health Assessment Questionnaire to measure functional disability. RESULTS: The 12 month resistance training in EG led to statistically significant mean increases of 22-35% in all muscle groups examined. CG patients were also able to increase their strength to some degree (3-24%), but at the end of the study strengths in CG were significantly lower than in EG. By the end of the study lumbar spine BMD had changed by +0.19% (4.24) in EG and by -1.14% (4.36) in CG. The corresponding changes of femoral BMD were +1.10% (3.71) and -0.03% (3.58). The changes in BMD were minor and statistically not significant in both groups. However, femoral BMD was found to be decreased among those patients treated periodically with oral glucocorticoids (n = 15, 3 subjects from EG and 12 from CG) compared with changes in BMD among those not treated with systemic glucocorticoids (n = 50). CONCLUSION: Minimally supervised strength training resulted in significant improvements in muscle strength without detrimental effects on disease activity. The detected annual changes in central BMD were minor and statistically insignificant in both groups. Special attention should be focused on those patients with RA with high disease activity and concomitant glucocorticoid treatment. PMID- 10381041 TI - Modeling the lifetime costs of rheumatoid arthritis. AB - OBJECTIVE: To develop an analytical approach for estimating the lifetime costs of rheumatoid arthritis (RA) using existing population based cross sectional data, and to use this estimate to describe the potential cost-effectiveness of bone marrow transplantation for RA. METHODS: Estimates of arthritis related costs (direct, indirect, and nonmedical) and mortality were obtained from previously assembled population based cohorts. A mathematical model was designed defining 25 hypothetical ratios (RA/NA) representing the proportionate excess cost of RA each year for the 25 years following a diagnosis of RA. Using age and sex-specific cost estimates, we then simulated a vector of 25 ratios 1000 times. Each age and sex-specific randomly generated variable was converted to an estimated dollar amount (in 1995 dollars US) of excess cost attributable to RA. All dollar amounts were discounted by 3% per year. Finally, each vector of 25 discounted dollar amounts was summed to yield an estimate of the total excess medical costs in 1995 dollars for the first 25 years of a person's lifetime following a diagnosis of RA. Because not every one of these hypothetical individuals would be expected to live all 25 years, we used the standardized mortality ratio for an individual with RA (from our inception cohort) and multiplied it by the age-specific 1990 mortality rates for Minnesota whites to estimate how many of the 1000 randomly generated hypothetical individuals could be expected to die during each of the 25 years. For these, the summation of estimated cost was truncated at the death year. This process yielded, for each age and sex, a sample of 1000 sums of 25 (or fewer) excess costs all in terms of 1995 dollars that correspond to the excess cost during the first 25 years after an RA diagnosis, adjusted for differential survival among patients with RA compared to nonarthritic controls. The distribution of these sums thus represented a distribution of the 1995 dollars that could be saved if RA could be "cured" soon after incidence. RESULTS: Our simulation analyses indicated that the median lifetime incremental costs of RA range roughly from ,$61,000 to $ 122,000. Incremental costs were higher for younger individuals compared to older individuals and were consistent over all percentiles and age groups. No systematic relationship between the incremental costs of females with RA compared to males was identified. CONCLUSION: These data suggest that interventions such as autologous bone marrow transplantation, which has recently been estimated to cost roughly $60,000, may be cost saving if they eliminate the downstream incremental costs of RA. PMID- 10381042 TI - Double blind, randomized, placebo controlled clinical trial of methotrexate in systemic lupus erythematosus. AB - OBJECTIVE: To evaluate the capacity of methotrexate (MTX) to control mild activity of systemic lupus erythematosus (SLE), and to evaluate the capacity of MTX to reduce steroid requirement, as well as to evaluate the side effects of MTX in patients with SLE. METHODS: A prospective, randomized, controlled, double blind trial. Forty-one patients with SLE began and 37 completed the 6 months of study. The mean disease duration was 82.5 months. Twenty patients received MTX 15 20 mg/week (MTX group) and 21 received placebo (PL group). The dose of prednisone was maintained, increased, or reduced after the first month, according to monthly clinical and laboratory evaluation. Dose of prednisone, SLE Disease Activity Index (SLEDAI) scores, the score by visual analog scale (VAS) for articular pain, and laboratory results were recorded monthly. Both groups were homogeneous and comparable for clinical manifestations and laboratory results. RESULTS: Two placebo patients dropped out due to severe flare of disease requiring hospitalization, and 2 patients taking MTX dropped out due to side effects (one with pulmonary tuberculosis, one with urticaria and severe dyspepsia). Thirty seven patients (18 MTX and 19 PL) completed the study. At the end of the study 16 PL patients and one MTX patient presented articular complaints (p < 0.001). VAS scores for pain were significantly higher in the PL group than in the MTX group after the first month of study. Sixteen PL patients and 3 MTX patients presented cutaneous lesions after 6 months of treatment (p < 0.001). At the end of the study 4 MTX patients and 11 PL patients presented hypocomplementemia (p < 0.001). Mean SLEDAI scores in PL patients were significantly higher than in MTX patients at Months 3, 4, 5, and 6. It was possible to decrease the prednisone dose for 13 MTX patients during the study but for only one PL patient (p < 0.001). Fourteen MTX patients (70%) presented side effects, mainly dyspepsia and increase in hepatic enzyme serum levels, and 3 PL patients (14%) presented dyspepsia. CONCLUSION: MTX 15 to 20 mg/week for 6 months was effective in controlling cutaneous and articular activity of SLE and permitted prednisone dose reduction. At these doses MTX presented frequent but mild side effects that did not result in drug discontinuation in the majority of patients. PMID- 10381043 TI - Determinants of bone mass in systemic lupus erythematosus: a cross sectional study on premenopausal women. AB - OBJECTIVE: To evaluate bone mineral density (BMD) in young ambulatory female patients with systemic lupus erythematosus (SLE) and to assess the influence of disease related variables and use of corticosteroids. METHODS: Lumbar and femoral BMD were measured by dual x-ray absorptiometry (DXA) in 84 premenopausal patients with SLE (age 30.5+/-7.5 years). All patients were receiving corticosteroids at the time of the study. Variables evaluated were: disease duration, clinical pattern, disease activity (SLEDAI), cumulative damage index (SLICC/ACR), current and cumulative prednisone dose, duration of steroid treatment, and use of immunosuppressive agents. Osteoporosis was defined as a t score below 2.5 SD compared to a reference population of healthy women in at least one region of measurement. RESULTS: Vertebral and femoral BMD were significantly lower in patients with SLE than in age matched controls. Osteoporosis was detected in 22.6% of patients. No significant differences in BMD were detected between patients according to clinical pattern or activity index, whereas patients with damage index > 0 (n = 46) had a significantly lower BMD at both the lumbar (p = 0.008) and the femoral (p = 0.05) level. Compared with non-osteoporotic patients with SLE, women with osteoporosis had similar age, lower body mass index, significantly longer disease duration (p < 0.0001), higher cumulative steroid intake (p < 0.006), and higher SLICC/ACR score (p < 0.01). Stepwise logistic regression analysis showed that disease duration is independently associated with osteoporosis (OR 1.2 for each year of disease, 95% CI 1.07-1.33). Since disease duration and duration of steroid treatment were highly correlated, a new stepwise logistic model was run without disease duration, which revealed that prednisone was associated with an increased risk for osteoporosis (OR 1.16 for each year of treatment, 95% CI 1.05-1.29). CONCLUSION: Osteoporosis is a frequent feature in young patients with SLE. Disease duration is associated with an increased risk for osteoporosis, but the role of glucocorticoid treatment seems to be crucial. Steroid exposure was the only treatment related variable exerting an influence on the development of osteoporosis. PMID- 10381044 TI - Can health utility measures be used in lupus research? A comparative validation and reliability study of 4 utility indices. AB - OBJECTIVE: To assess validity and reliability of 4 utility indices in patients with systemic lupus erythematosus (SLE). METHODS: Twenty-five patients with stable SLE underwent assessment of disease activity [Systemic Lupus Disease Activity Measure (SLAM-R) and SLE Disease Activity Index (SLEDAI)] and damage [Systemic Lupus Collaborating Clinics/American College of Rheumatology Damage Index (SLICC/ACR DI)] and completed a health survey [Medical Outcome Survey Short Form-36 (SF-36)] and 4 utility measures: the visual analog scale (VAS), the time trade-off (TTO), the standard gamble (SG), and the McMaster Health Utilities Index Mark 2 (HUI2). To assess validity, Pearson's correlations were calculated between the SF-36 subscales and the utility measures. To assess reliability, intraclass correlations or kappa coefficients were calculated between first and second assessments, performed from 2 to 4 weeks apart, in patients without important clinical change in disease activity. RESULTS: Disease activity from a SLAM-R varied from 0 to 14 (median = 4) and SLEDAI from 0 to 18 (median = 0). All subscales of the SF-36 correlated well with the VAS [lowest r = 0.56, 95% confidence interval (CI) (0.17, 0.80)] and poorly with the SG [maximum r = 0.41, CI (-0.01, 0.70); minimum r = 0.10, CI (-0.32, 0.50)]. The subscales of bodily pain (r = 0.56), mental health (r = 0.45), physical functioning (r = 0.62), role emotional (r = 0.47), social functioning (r = 0.49) and vitality (r = 0.44) correlated significantly with TTO. All subscales correlated significantly [lowest r = 0.48, CI (0.09, 0.75)] with the HUI2 index of pain. Intraclass correlations for the VAS (ICC = 0.67) and TTO (ICC = 0.60) were good. They were fair for the SG (ICC = 0.45). The kappa coefficient was poor (0.32) for the HUI attribute of pain, but varied from fair (0.46) to excellent (0.88) for the remaining attributes. Regression analysis showed that a model incorporating the SLAM-R value and SF-36 subset of mental health was a good predictor of VAS and TTO utility measures. CONCLUSION: The VAS, TTO, and to some extent, the HUI2, when compared with the SF-36 health survey, are valid and reliable measures to assess health related quality of life in a group of patients with SLE and may be useful for future research in this population. On the basis of these results the usefulness of the SG is questionable in these patients. PMID- 10381045 TI - The prevalence of hyperprolactinemia in patients with primary Sjogren's syndrome. AB - OBJECTIVE: To assess the prevalence of hyperprolactinemia in 55 patients with primary Sjogren's syndrome (SS), and its clinical significance. METHODS: Concentrations of serum prolactin (PRL) were determined in 55 consecutive patients with primary SS and 110 controls by a fluoroimmunometric assay in a prospective case-control design. RESULTS: The 55 patients with primary SS had higher serum PRL than 110 matching controls (271.5 vs 205.9 mIU/l; p < 0.02), and this difference was most evident in patients diagnosed before the age of 45 years (374.8 vs 245.5 mIU/l; p < 0.05), a patient population characterized by active immunological disease. Serum PRL did not correlate to disease duration, serum immunoglobulin, autoantibodies, or focus score in biopsies from minor salivary glands, but did correlate to score for internal organ disease (r = 0.33, p < 0.05). Two patients were diagnosed as having primary SS 12 years after hyperprolactinemia was first detected, and both patients had aggressive primary SS as indicated by extraglandular manifestations. One of the patients developed primary SS after being treated with bromocriptine, an inhibitor of PRL synthesis, for 12 years. CONCLUSION: Patients with primary SS have moderately increased levels of serum PRL, especially evident in patients diagnosed at a young age with active immunological disease. Serum PRL is correlated to index for internal organ disease, and primary SS may be preceded by hyperprolactinemia for many years. PMID- 10381046 TI - Survivorship in a population based cohort of patients with Sjogren's syndrome, 1976-1992. AB - OBJECTIVE: Sjogren's syndrome (SS) has been associated with development of lymphoid malignancies and other significant medical complications, but the effect of SS on survival in a population based sample has not been reported. We analyzed survival in an incidence cohort of patients diagnosed with SS in residents of Olmsted County, Minnesota, USA, between 1976 and 1992. METHODS: All records of physician diagnosed SS were reviewed, as well as all records from patients diagnosed with xerostomia and keratoconjunctivitis sicca, and records of patients with rheumatoid arthritis (RA) and systemic lupus erythematosus. The case definition for SS required 2 of 3 criteria: keratoconjunctivitis sicca, xerostomia, or serologic abnormality. Confounding illnesses were excluded. All patients were white. RESULTS: Of the 74 cases of SS identified, 50 (67%) had primary SS and 24 (33%) secondary SS. An average of 7.2 years of followup was available for patients with primary SS and 9.9 years for patients with secondary SS. Compared with the general population, patients with SS had increased mortality (p = 0.04). When patients with primary and secondary SS were studied separately, increased mortality was found in patients with secondary SS (p < 0.005) but not primary SS (p = 0.86). CONCLUSION: In this population based cohort, patients with primary SS did not have increased mortality. However, mortality may have been increased in patients with secondary SS, the majority of whom had RA. PMID- 10381047 TI - Cerebral white matter lesions in primary Sjogren's syndrome: a controlled study. AB - OBJECTIVE: To determine the prevalence of neurological and magnetic resonance imaging (MRI) abnormalities in a well defined population of unselected patients with primary Sjogren's syndrome (SS) and age and sex matched healthy patients. METHODS: Thirty patients with SS and 29 age and sex matched controls were examined by a neurologist and subsequently underwent MRI scanning with a 1.0 Tesla Siemens Impact MR scanner. Scans were graded by a neuroradiologist blinded to the clinical status of each subject. The number and location of white matter lesions > 3 mm in long axis (to exclude non-specific perivascular changes) were recorded for each subject. RESULTS: There was a significant increase in lesions detected by MRI in SS patients versus controls (p = 0.02) including deep white matter lesions (p = 0.03) and subcortical white matter lesions (p = 0.02). The presence of white matter lesions did not correlate with serum IgG or rheumatoid factor levels, or with presence of anticardiolipin antibodies. No subjects had symptoms or signs of serious neurological disease including multiple sclerosis, and corpus callosal lesions commonly seen in multiple sclerosis were notably absent in this study. CONCLUSION: Cerebral white matter lesions detected by MRI are more frequent in patients with primary SS than control subjects, yet do not appear to be associated with significant clinical manifestations. Although the pathological nature of these lesions is yet to be defined, their presence should not be over-interpreted. PMID- 10381048 TI - Presence of antibodies against Helicobacter pylori and its heat-shock protein 60 in the serum of patients with Sjogren's syndrome. AB - OBJECTIVE: Helicobacter pylori infection elicits a local and systemic immune response against bacterial antigens, including a heat-shock protein of 60 kDa (HSP60). The homology between microbial and human HSP suggests that the immune response to bacterial HSP may play a role in the pathogenesis of autoimmune disorders. Since gastric involvement and H. pylori have been reported in Sjogren's syndrome (SS), we investigated the prevalence of antibodies against H. pylori and its specific HSP60 in sera from patients with SS. METHODS: Four groups of patients were studied. Group 1, 34 patients with primary SS (pSS); Group 2.19 patients with secondary SS; Group 3, 22 patients with various autoimmune diseases and Group 4, 43 healthy controls. Serum IgG levels against HSP60 were determined by an ELISA using recombinant full length HSP60 expressed in Escherichia coli, as the antigen. To confirm the H. pylori infection, a commercial ELISA was used. RESULTS: Out of 34 patients in Group 1, 27 (79.4%) and 30 (88.2%) had antibodies against H. pylori and its HSP60, respectively. The prevalence was significantly higher than that found in Group 3 (18.2%, p < 0.0001 and 27.3%, p < 0.0001) and in Group 4 (48.8%, p < 0.005 and 37.2%, p < 0.0001) but not than that of Group 2 (48.8% and 37.2%). If the prevalence of patients either positive or negative for both antibodies was considered, a statistically significant difference was found between Group I and respectively Groups 3 and 4. CONCLUSION: The hypothetical role of HSP60 in the development of the immune response both in pSS and secondary SS seems strictly linked to the prevalence rate of H. pylori infection. PMID- 10381049 TI - Sera from patients with scleroderma inhibit fibroblast micromotions monitored electrically. AB - OBJECTIVE: To investigate scleroderma fibroblast behavior, and the effect of different human sera on fibroblast behavior, we established a model using the newly developed electrical biosensor, electrical cell-substrate impedance sensing (ECIS). METHODS: Cellular locomotion, defined as cellular micromotion, measured by ECIS indicates the dynamic vertical motion of a given group of cells. The junctional resistance (Rb) between adjacent cells and the average height (h) between basal cell surface and substratum derived from ECIS were quantified for dermal fibroblasts obtained from patients with scleroderma and normal controls. The cellular micromotions of both scleroderma and normal control fibroblasts were compared in the presence of different human sera, including those from patients with scleroderma, systemic lupus erythematosus (SLE), and Sjogren's syndrome (SS) and from normal individuals. RESULTS: The Rb and h levels for scleroderma and normal fibroblasts were 2.7 omega.cm2, 150 nm and 0.8 omega.cm2, 303 nm respectively. The micromotions of scleroderma fibroblasts were more active than those of normal fibroblasts. Sera from patients with scleroderma can inhibit the micromotions of normal fibroblasts but not those of scleroderma fibroblasts, while sera from patients with SLE and SS have no inhibitory effect on either normal or scleroderma fibroblast micromotions. CONCLUSION: We have demonstrated the previously unrecognized characteristics of dermal fibroblasts and sera derived from patients with scleroderma. It is possible that in vivo activities cause scleroderma fibroblasts to display active cellular micromotions, while sera from patients with scleroderma inhibit the micromotions of normal fibroblasts. The use of ECIS technology has also provided a new approach to the study of scleroderma. PMID- 10381050 TI - Favorable role of interleukin 10 in patients with polymyalgia rheumatica. AB - OBJECTIVE: To determine the relationship between the antiinflammatory molecule interleukin 10 (IL-10) and disease symptoms, IL-1beta, tumor necrosis factor (TNF), and IL-1 receptor antagonist (IL-1ra) in patients with polymyalgia rheumatica (PMR). METHODS: In 102 patients with PMR, we determined the severity of the disease by the presence of typical clinical symptoms (symptom score with a maximum of 10 points). IL-10, IL-1beta, TNF, and IL-1ra were measured in all patients and 31 age matched healthy controls by enzyme immunometric assays. RESULTS: Compared to patients with elevated serum levels, patients with normal serum levels of IL-10 (below the mean + 3 SD of controls, 7.79 pg/ml) more often had adynamia (p = 0.045), bilateral muscular pain in shoulders, upper arms or neck (p = 0.045), bilateral muscular pain in the pelvic girdle (p < 0.001), headache (p = 0.014), morning stiffness (p < 0.001), symptoms of depression (p = 0.013), and initial weight loss (p = 0.011), and had a higher symptom score (5.5+/-0.4 vs 3.7+/-0.3; p < 0.001). The overall symptom score correlated negatively with IL-10 serum levels (Rrank = -0.421, p < 0.001). IL-10 correlated negatively with IL-1beta (p = 0.013) and TNF-alpha (p = 0.039). The association between elevated serum levels of IL-10 and low serum levels of IL-1beta and TNF was observed only in patients with corticosteroid treatment. In these patients, elevated serum levels of IL-10 were positively associated with an increased ratio of IL-1ra to IL-1beta. CONCLUSION: Elevated serum levels of IL-10 were associated with a more mild form of PMR. This study indicates a favorable role of IL-10 in patients with PMR. PMID- 10381051 TI - The spectrum of polymyalgia rheumatica in northwestern Spain: incidence and analysis of variables associated with relapse in a 10 year study. AB - OBJECTIVE: To examine patients presenting with polymyalgia rheumatica (PMR) during a 10 year period in Northwestern Spain and to assess disease incidence and the frequency of relapses in patients diagnosed as having either isolated ("pure") PMR or PMR associated with giant cell arteritis (GCA). METHODS: Clinical records of patients with PMR diagnosed at the Hospital Xeral Lugo from January 1987 through December 1996 were reviewed. Patients with PMR were categorized into 2 subgroups depending on the presence or absence of associated GCA. Other conditions presenting with polymyalgia symptoms or mimicking isolated PMR were excluded. These patients were followed from the time of diagnosis until either patient's death or January 1, 1998. RESULTS: One hundred eighty-five patients were studied. The average annual incidence rates of the total group of PMR and isolated PMR were 18.67 x 10(-5) and 13.52 x 10(-5), respectively, in a population aged 50 years or older. Relapses were frequent in both isolated PMR and PMR associated with GCA. In general, they occurred when the dose of prednisone was < 7.5 mg/day or it had been discontinued. Rate of steroid tapering was significantly higher in patients with isolated PMR who had relapses. Patients with HLA-DRB1*0401 had a higher frequency of relapses. As reported in patients with PMR associated with GCA, the adjusted mortality rate in patients with isolated PMR showed no difference compared to the Spanish population aged 50 years or older. CONCLUSION: In Northwestern Spain, PMR is a nonfatal disease with a low incidence and frequent relapses. A possible influence of HLA-DRB1*04 alleles in the development of more severe disease, with greater tendency to relapses, is suggested. PMID- 10381052 TI - Polymyalgia rheumatica with low erythrocyte sedimentation rate at diagnosis. AB - OBJECTIVE: To determine the frequency and clinical characteristics of polymyalgia rheumatica (PMR) with low erythrocyte sedimentation rate (ESR) at diagnosis in a community based cohort of 232 patients. METHODS: A retrospective review of medical records of all the patients with a diagnosis of PMR in Olmsted County, Minnesota, seen and followed over a 22 year period, from 1970 through 1991. RESULTS: Seventeen (7.3%) patients had ESR < 40 mm/h at diagnosis. The findings and outcome in these patients were compared with the others in the group. There was no difference in sex or age between the 2 groups. Both groups had the same delay to diagnosis, typical gradual onset of the disease, the same frequency of both proximal and distal stiffness/pain, and the same frequency of synovitis. Systemic features were less frequent in the low ESR group than in the high ESR group (59 vs 81%, p = 0.05). Mean ESR in the low ESR group was 26+/-9 mm/h versus 74+/-24 mm/h in the high ESR group. The mean hemoglobin concentration was significantly lower (p = 0.0015) in the high compared to the low ESR group (12.2+/-1.4 g/dl versus 13.3+/-1.3 g/dl). The frequency of positive temporal artery biopsy and of diagnosed giant cell arteritis was the same in the 2 groups. Initial response to therapy, frequency of relapses, number of patients going into remission, time to remission, and daily dose of steroids were the same in both groups. CONCLUSION: Other than more frequent systemic symptoms, our population of patients with PMR and low ESR at diagnosis had similar clinical characteristics and course of disease as patients with high ESR at diagnosis. PMID- 10381053 TI - A proposal for the classification of patients for clinical and experimental studies on reactive arthritis. AB - OBJECTIVE: To propose classification criteria for patients entering clinical and basic studies on reactive arthritis (ReA). METHODS: From a MEDLINE search of articles published between 1980 and 1996, we identified reports on HLA-B27 related ReA and Reiter's syndrome as study groups and analyzed the items that constituted the diagnostic, classification, and inclusion (or entry) criteria of patients. We developed disease categories that constituted our classification proposal. RESULTS: We reviewed 175 articles containing 110 study groups of patients with ReA and 94 with Reiter's syndrome. Most articles (89.7%) relied on arthritis for diagnosis, but only 48.0% relied on infection. Only 22.5% of articles used published criteria for diagnosis. Articles including a bacterial name to further describe a group of patients with ReA relied on cultures at the site of infection, serum antibodies, or both to confirm the diagnosis. There were inter/intra-group variations and overlapping of diagnostic criteria, at least 32 different terms referring to ReA or Reiter's syndrome, and 6 patterns of disease. According to these data, we propose 3 categories of disease for patients entering clinical and basic studies on ReA: probable ReA (2 subgroups); definite ReA triggered by bacteria (2 subgroups); and bacterial-associated undifferentiated oligoarthritis or spondyloarthropathy. CONCLUSION: This proposal provides a rationale for reducing the heterogeneity found in criteria for including patients with ReA in research and to facilitate scientific communication. In contrast to diagnostic criteria, this proposal does not restrict the study population to a minority of patients, but allows the investigator to include several forms of disease and to analyze results according to different categories. PMID- 10381054 TI - Antiarthritic activity of soluble tumor necrosis factor receptor type I forms in adjuvant arthritis: correlation of plasma levels with efficacy. AB - OBJECTIVE: To determine the importance of tumor necrosis factor (TNF) in the pathogenesis of adjuvant disease in rats and to determine plasma levels of recombinant soluble TNF receptor type I (sTNF-RI) necessary for efficacy, to project dosing for human clinical trials. METHODS: Rats with adjuvant arthritis were treated by continuous infusion with sTNF-RI forms to maintain blood levels of this TNF-alpha inhibitory protein. In addition, rats were given bolus injections of polyethylene glycol linked sTNF-RI and efficacy and plasma levels were determined. Effects of treatment in the rats were monitored by sequential volume or diameter measurement of ankle joints, final paw weights, and histologic evaluation of ankle joints, with particular emphasis on bone erosive lesions. RESULTS: In all studies and regardless of dosing methodology (bolus vs continuous infusion), minimal plasma levels for efficacy were in the 0.3-0.5 microg/ml range. Higher plasma levels resulted in greater efficacy, with maximal effects achieved when plasma levels were in the 5 microg/ml range. Beneficial effects of treatment were seen on body weight, paw swelling, splenomegaly, hepatomegaly, and bone resorption. CONCLUSION: TNF-alpha is an important mediator of all aspects of rat adjuvant disease including both the destructive processes in the joints as well as the systemic manifestations of adjuvant disease. Studies using various forms of sTNF-RI consistently show that plasma levels of 0.3-0.5 microg/ml are required for minimal efficacy and that higher plasma levels show dose-responsive enhanced efficacy. PMID- 10381055 TI - Amelioration of type II collagen induced arthritis in rats by treatment with sodium diethyldithiocarbamate. AB - OBJECTIVE: Sodium diethyldithiocarbamate (Ditiocarb, DDTC), which is used in the treatment of heavy metal poisoning, effectively inhibits NF-kappaB activation and cytokine secretion in vitro. To investigate the antiinflammatory and immunosuppressive potency of DDTC, we examined its influence on the course of collagen induced arthritis in rats. METHODS: Arthritis was induced in female DA rats by injection of rat collagen type II emulsified in incomplete Freund's adjuvant into the tail base. After onset of arthritis, the animals received DDTC or vehicle by intraperitoneal injections or subcutaneous infusion using osmotic pumps. Disulfiram, which is cleaved into DDTC within the gastrointestinal tract, was administered orally via gastric gavage. The course of arthritis was followed by clinical scoring and measurement of joint swelling. RESULTS: Collagen induced arthritis was significantly ameliorated by intraperitoneal injection (2 x 300 mg/kg/day) and subcutaneous infusion (120 mg/kg/day) of DDTC and by enteral administration of disulfiram (200 and 300 mg/kg/day). CONCLUSION: Dithiocarbamates may provide an effective new approach for the treatment of arthritis and other inflammatory diseases. PMID- 10381056 TI - Field test of the reproducibility of automated measurements of medial tibiofemoral joint space width derived from standardized knee radiographs. AB - OBJECTIVE: To estimate the reproducibility of computerized measurements of minimum joint space width (JSW) in the medial tibiofemoral compartment in knee radiographs (semiflexed AP view) obtained from clinical radiology units. METHODS: Technologists from 5 clinical radiology units were trained in the performance of the fluoroscopically assisted semiflexed AP knee examination. Each of 44 subjects (34 with knee osteoarthritis, OA, 10 with bilaterally normal knees) were examined within 7 days in 2 of the 5 units. The examination in each unit was repeated 1 week later. Minimum JSW was measured on digitized radiographic images with computer software that corrected for radiographic magnification. RESULTS: Despite ongoing quality control by technologists, 11% of radiographs were flawed with respect to the protocol standard for knee rotation and 36% with respect to the standard for knee flexion. The standard error of measurement (SEm) of JSW in 174 knees that were examined twice in the same unit was 0.32 mm (SEm = 0.25 mm for the subset of 76 paired radiographs with uniformly high quality). The overall between-unit SEm was 0.45 mm. Within-unit, but not between-unit, precision was related to the technical quality of the radiographs. Precision was unrelated to subject age, sex, race, weight, and radiographic severity of knee OA. CONCLUSION: The within-unit precision of JSW measurements from all pairs of semiflexed views (irrespective of technical quality) represented a notable improvement over that observed in radiographs with flawed knee rotation or flexion (as would be the case in conventional extended knee views). In future applications of this technique, assurance of technical quality by an independent observer should result in a level of measurement precision that will permit the design of clinical trials of disease modifying OA drugs with fewer subjects and/or shorter duration of treatment than is possible with conventional knee radiography. PMID- 10381057 TI - Effect of antiinflammatory drugs on COX-1 and COX-2 activity in human articular chondrocytes. AB - OBJECTIVE: To study the effect of steroidal and nonsteroidal antiinflammatory drugs (NSAID) on cyclooxygenase (COX-1 and COX-2) activity in human articular chondrocytes. METHODS: Chondrocytes were isolated from articular cartilage of donors with no articular disease. Unstimulated and interleukin 1 (IL-1) stimulated chondrocytes were used as models to study the effects of drugs on COX 1 and COX-2. Cells were incubated with vehicle or drugs; supernatants were removed and the level of prostaglandin E2 (PGE2) in each sample was determined by enzyme immunoassay. IC50 were calculated from the reduction in PGE2 content by different concentrations of the test substance by linear regression analysis. RESULTS: COX- mRNA was detected in unstimulated cells, but stimulation with IL-1 for up 12 h did not modify the levels of COX-1 mRNA. In contrast, COX-2 mRNA was not detectable in unstimulated cells, but it was induced by IL-1. Dexamethasone inhibited COX-2 mRNA expression induced by IL-1. COX-2 protein levels correlated with mRNA expression. Dexamethasone was the strongest drug inhibitor of COX-2 (IC50 = 0.0073 microM). However, it did not inhibit COX-1 activity. Among all NSAID tested, meloxicam and aspirin were the least potent inhibitors of COX-1 (IC50 = 36.6 microM and 3.57 microM, respectively). Indomethacin and diclofenac were the most potent inhibitors of COX-1 (IC50 = 0.063 microM and 0.611 microM, respectively) and COX-2 isoforms (IC50 = 0.48 microM and IC50 = 0.63 microM, respectively). Meloxicam was a more potent inhibitor of COX-2 (IC50 = 4.7 microM) than aspirin (IC50 = 29.3 microM) and similar to piroxicam (IC50 = 4.4 microM). Among all drugs tested dexamethasone showed the greatest selectivity for COX-2 and meloxicam was the NSAID with the best COX-2/COX-1 ratio (r = 0.12). Aspirin and piroxicam were about 8 times more active against COX-1 than COX-2, indomethacin was 7 times more active, and diclofenac was an equipotent inhibitor of COX-1 and COX-2. CONCLUSION: We found that COX-1 and COX-2 isoforms are expressed in human chondrocytes at rest and in IL-1 stimulated cells, respectively. Antiinflammatory drugs have different capacities to inhibit COX enzyme in human articular chondrocytes. PMID- 10381058 TI - When did "osteo-arthritis" become osteoarthritis? PMID- 10381059 TI - Decreased capillary density in juvenile dermatomyositis and in mixed connective tissue disease. AB - OBJECTIVE: To assess whether quantitative capillary microscopy is a useful tool to evaluate capillary abnormalities in children with connective tissue diseases. METHODS: Eight children with juvenile dermatomyositis (JDM), 6 with mixed connective tissue disease (MCTD) and 23 healthy children were investigated with computer based quantitative capillary microscopy. Median disease duration was 1 year among JDM (1-4) and 3 years (1-7) among MCTD. RESULTS: Capillary density was decreased in JDM [median 2.5 (1.4-4.3) loops/mm (p < 0.001)] and in MCTD [median 5.0 (4.1-7.0) loops/mm (p < 0.05)] compared to healthy controls [median 6.8 (5.3 8.0) loops/mm]. Median capillary loop area was increased in JDM [median 8.5 (3.0 15.8) 10(-3) mm2 (p < 0.001)] and in MCTD [median 4.5 (3.0-6.0) 10(-3) mm2 (p < 0.02)] compared to controls [median 2.5 (1.0-4.0) 10(-3) mm2]. CONCLUSION: Quantitative nailfold capillary microscopy is a sensitive indicator of JDM. In MCTD this technique is less discriminative. PMID- 10381060 TI - Pulmonary function in children affected by juvenile spondyloarthropathy. AB - OBJECTIVE: Pleuropulmonary involvement in adult spondyloarthropathy (SpA) has been thoroughly investigated. SpA is usually detected by conventional radiology as fibrosis of the upper lobes in about 30% of asymptomatic patients. Pulmonary function tests (PFT) reveal decreased vital capacity and total lung capacity, as well as increased residual volume. Juvenile SpA (JSpA) is a rare clinical condition, and no extensive investigations have been carried out on pulmonary involvement in JSpA. We studied prevalence and features of PFT alterations in patients with JSpA over a 2 year followup and analyzed the relationship between PFT and disease duration, disease activity, and chest and spine mobility. METHODS: Eighteen patients with JSpA, with no clinical signs of lung disease and normal chest radiographs, underwent PFT--static and dynamic volumes, diffusing capacity for carbon monoxide (DLCO), at enrollment (T0), at 12 months (T1), and at 24 months later (T2). Disease activity was defined at each investigation by clinical and hematological data. RESULTS: Significant functional lung impairment was detected in 33% of patients (reduced forced vital capacity in 22% and DLCO in 11%). No significant change in the prevalence and features of PFT alterations was detected at T1 and T2; no relationship was found between PFT and duration, activity, and clinical scores of the disease. CONCLUSION: Thirty-three percent of JSpA patients without clinical symptoms and no radiological findings of lung involvement show PFT alterations, mainly characterized by a restrictive pattern. No progression or modification in PFT developed over 2 years. No correlation was found between PFT and disease duration, activity, and clinical scores. PMID- 10381061 TI - Childhood Churg-Strauss syndrome. AB - Churg-Strauss syndrome or allergic granulomatosis and angiitis is a vasculitis that is found in adults, but is extremely rare in children. We describe a 14-year old boy who presented with prolonged fever, weight loss, sinusitis, myalgia and arthralgia, testicular pain, pulmonary infiltrations, pericardial effusion, peripheral neuropathy, and eosinophilia. Muscle biopsy showed necrotizing arteritis with eosinophil infiltration. His clinical course was complicated by several seizures secondary to cerebral vasculitis and severe asthma, resulting in death. The clinical features and outcomes of childhood Churg-Strauss syndrome are reviewed. PMID- 10381062 TI - Complete heart block as a rare complication of treatment with chloroquine. AB - Antimalarials are well established disease modifying antirheumatic drugs. A rare and underappreciated treatment difficulty is cardiac complication, in particular conduction disturbances. We describe 2 more patients that developed complete heart block after high dose, longterm treatment. Patient 1, a 73-year-old woman with longstanding rheumatoid arthritis, had taken chloroquine (250 mg/day) for 12 years when she developed complete heart block requiring a permanent pacemaker. Patient 2, a 40-year-old woman with discoid lupus erythematosus, was taking chloroquine from 1979 until 1996. Depending on the clinical disease activity, she intermittently increased the dose from 250 to 750 mg/day. In 1994, she developed complete heart block and a permanent pacemaker had to be implanted. Intensive investigations in both cases did not reveal another underlying cause for conduction disturbances; the atrioventricular block was probably due in both cases to chloroquine related cardiac toxicity. This toxicity seems to be restricted to longterm, high dose treatment; however, it should be kept in mind in patients with preexisting conduction disturbances during longterm treatment. PMID- 10381063 TI - Successful treatment of methotrexate induced nodulosis with D-penicillamine. AB - Accelerated nodulosis is a recognized complication of methotrexate (MTX) therapy in rheumatoid arthritis (RA). We describe 3 patients with accelerated nodulosis treated with D-penicillamine (D-Pen) while continuing MTX. The combination of D Pen with MTX therapy resulted in regression of subcutaneous nodules in all patients, disappearance of pulmonary nodules in one patient, and resolution of vasculitic lesions in 2 patients. Clinical response was observed within the first few weeks of therapy and usually required moderate doses (500 mg/day). Our observations suggest that addition of D-Pen to MTX therapy can be an alternative therapeutic option for accelerated nodulosis in patients with RA. PMID- 10381064 TI - Systemic lupus erythematosus and Castleman's disease. AB - Lymphadenopathy is a common finding in systemic lupus erythematosus (SLE), yet lymphoid malignancy is rare. Typically, adenopathy associated with SLE responds to glucocorticoid therapy. We evaluated a patient with a diagnosis of SLE who had progressive lymphadenopathy despite receiving aggressive corticosteroid therapy for SLE associated thrombocytopenia. Histopathology initially revealed an aggressive plasmacytosis characteristic of Castleman's disease (CD). CD, or angiofollicular hyperplasia, is a rare lymphoproliferative neoplasm that has features overlapping many autoimmune diseases. This disorder should be considered in autoimmune diseases with unremitting or progressive adenopathy. PMID- 10381065 TI - Cogan's syndrome with refractory abdominal aortitis and mesenteric vasculitis. AB - Cogan's syndrome is a rare multisystem disease characterized by ocular inflammation, vestibuloauditory dysfunction, and vasculitis. We report a 26-year old Caucasian woman who died from Cogan's syndrome. Her case illustrates that patients with Cogan's syndrome can have abdominal aortitis and mesenteric vasculitis, and that the vasculitis can be refractory to methotrexate, cyclophosphamide, cyclosporine, and chlorambucil. PMID- 10381066 TI - Spinal cord granulomatous vasculitis: an unusual clinical presentation of sarcoidosis. AB - A 52-year-old Caucasian man presented with isolated manifestations of myelopathy. Cervical magnetic resonance imaging showed a focus of increased signal intensity at the posterolateral left half of C5 and C6. Biopsy of spinal cord revealed the presence of active vasculitis associated with noncaseating granulomas. The patient responded to the combination of methotrexate (MTX) and corticosteroid treatment. Low-dose MTX was an effective, steroid-sparing regimen. This is a rare condition of isolated spinal cord involvement associated with sarcoid vasculitis. PMID- 10381067 TI - Successful treatment of severe muscle necrosis with intravenous immunoglobulin. AB - Intravenous immunoglobulin (IVIG) is an effective treatment for a range of conditions believed to have an autoimmune basis, including inflammatory muscle disease. We describe a patient with muscle disease characterized by severe necrosis without clinical or biopsy evidence of inflammation or vasculitis, who responded to treatment with IVIG. PMID- 10381068 TI - Work disability and soft tissue rheumatism. PMID- 10381069 TI - Migrating monopredominant arthritis in children of Assyrian ancestry. PMID- 10381070 TI - Genetic epidemiology: caveat lector. PMID- 10381071 TI - Alternative therapies and alternative medicine. PMID- 10381072 TI - Anti-M protein antibody in post-streptococcal reactive arthritis. PMID- 10381073 TI - Juvenile dermatomyositis and celiac disease. PMID- 10381075 TI - Antineutrophil cytoplasmic antibodies in patients with spondyloarthropathies. A predictor of chronic and progressive inflammatory joint disease. PMID- 10381074 TI - Minocycline induced lupus and autoimmune hepatitis. PMID- 10381076 TI - Epstein-Barr virus associated arthritis. PMID- 10381077 TI - Magnetic resonance imaging in the early diagnosis of brucellar sacroiliitis. PMID- 10381078 TI - Severe pancytopenia caused by a single administration of low dose methotrexate in a patient undergoing hemodialysis. PMID- 10381079 TI - Glucose-6-phosphate dehydrogenase deficiency and osteoarthritis in men of Northern Sardinia. PMID- 10381080 TI - Numb chin syndrome as first sign of temporal arteritis. PMID- 10381081 TI - Gout and pseudogout chronobiology. PMID- 10381082 TI - Synthesis of a eukaryotic virus protein in a prokaryotic viral-cell system: production of the adenovirus type 2 fiber shaft fragment by a tightly regulated T7POL-M13 expression system. AB - The use of recombinant technology for the production of proteins of interest in biotechnology and medicine has grown immensely during the last decade. A major problem often encountered is the degradation of the recombinant product by host cell proteases. We developed a novel system based on the cloning and expression of an inducible phage T7 RNA polymerase into the main intergenic region of the phage M13-KO7. After infection of permissive bacterial strains with the engineered phage, the polymerase gene is transcribed, subsequently translated and gene fragments cloned under T7 promoter sequences are then transcribed. For the evaluation of this system, the gene encoding the shaft fragment of the adenovirus type 2 fiber was cloned into a pET 3a-based expression vector. Expression was demonstrated in a BL21(DE3) strain (containing one copy of the T7 RNA polymerase gene) and also in several F pili-containing bacterial strains only after infection with the proper bacteriophage. Several important parameters for heterologous gene expression in Escherichia coli were investigated. Different bacterial strains were evaluated for the production of the recombinant protein, following: the expression levels, the growth rates and the stability of the plasmid vector at different time intervals after induction. It was observed that the expression levels as well as division rates and plasmid stability differed between the different bacterial strains. The best expression levels were obtained when using the E. coli Top IOF' strain. Degradation was only observed in BL21(DE3) cells after 6 h of induction, whereas none of the F'-containing cells were shown to degrade the recombinant protein during the time of expression. This system, based on the T7 pol-M13 bacteriophage, was shown to be very tightly regulated for most of the bacterial strains evaluated with no expression before induction of the T7 RNA polymerase. PMID- 10381083 TI - A sensitive fluorescence in situ hybridization technique for detection of porcine reproductive and respiratory syndrome virus. AB - A sensitive fluorescence in situ hybridization (ISH) for detecting porcine reproductive and respiratory syndrome virus (PRRSV) RNA in viral infected tissue was developed using digoxigenin-labeled RNA probes targeted on the nucleocapsid gene of PRRSV. In situ RNA/RNA hybrids were detected with an anti-digoxigenin antibody alkaline phosphatase conjugate and further revealed with Fast Red TR salt/naphthol AS-MX phosphate using a fluorescent microscope. Viral nucleic acid was readily demonstrated within macrophages, known to be the major target of PRRSV. In addition, positively stained cells were found in the salivary gland and skin tissues which have not been reported to contain PRRSV infected cells before. In conclusion, the fluorescence ISH used in this study provides a fast and sensitive means for screening virus-infected tissues in which relatively few cells are affected. This advantage will be especially beneficial for studying viral persistence and for routine diagnosis of PRRSV infection. PMID- 10381084 TI - Detection and quantification of cell-free Epstein-Barr virus by polymerase chain reaction and subsequent DNA enzyme immunoassay. AB - Amplification by polymerase chain reaction and subsequent DNA enzyme immunoassay (DEIA) were employed to determine the number of genome equivalents of cell-free Epstein Barr virus (EBV) DNA in peripheral blood. The assay detected cell-free EBV DNA in the serum of 14 out of 18 patients with primary, productive EBV infection (sensitivity 77.7%) but not in healthy EBV carriers with latent infection (specificity 100%). Our assay has the potential for a clinical diagnostic tool to monitor patients at risk for EBV reactivation and productive infection with subsequent EBV-induced lymphoproliferative diseases. PMID- 10381085 TI - A competitive reverse transcription-polymerase chain reaction assay for quantitation of GB virus C/hepatitis G virus RNA that circumvents heteroduplex artifact. AB - The role of GB virus C (GBV-C)/hepatitis G virus (HGV) in hepatitis has been controversial. To investigate its possible pathogenicity and site(s) of replication, it is important to develop an accurate quantitative assay for both positive and negative strand GBV-C/HGV RNA. In this study, a competitive reverse transcription-polymerase chain reaction (RT-PCR) assay for both positive and negative strand GBV-C/HGV RNA quantitation was developed. In developing the quantitative assay, heteroduplex formation was repeatedly observed. A heterologous competitor RNA with GBV-C/HGV primer-binding sequences was introduced, and heteroduplex artifact was circumvented successfully. Two-hundred thirty-seven serum specimens were screened by RT-PCR for GBV-C/HGV RNA. Two of the 62 patients infected with chronic hepatitis C virus (HCV) were found to be positive for GBV-C/HGV RNA. None of the 50 other patients with no evidence of HCV infection and none of the 125 normal individuals were positive for GBV-C/HGV RNA. The sensitivity of RT-PCR was 3000 gE/ml (30 gE in RT-PCR). Alternate methods for residual DNA removal and its detection in synthetic RNA were introduced. A RT control containing no primer before PCR is necessary to evaluate the trace amounts of template DNA remaining in synthesized RNA. The method will differentiate reliably between positive and negative strand RNAs up to a 10(4) fold difference in titer. The positive and negative strand GBV-C/HGV RNAs were detected in one patient by RT-PCR and hybridization analysis, and the strand titer was determined by RT-PCR. PMID- 10381086 TI - Original reverse transcription polymerase chain reaction method to obtain the full-length cDNA of rice tungro spherical virus. AB - A two-step reverse transcription reaction combined with long PCR was developed in order to obtain the full-length cDNA from the 12.2 kbp genomic RNA of rice tungro spherical virus. A first step reverse transcription, performed at 45 degrees C using a reverse transcriptase deprived of RNase H activity, allowed the synthesis of a nearly full-length cDNA of 11.7 kbp. A second step reaction, carried out at 65 degrees C using a thermostable polymerase, was necessary to destabilise secondary structures present at the 5' extremity of the RNA template which hampered the reverse transcription reaction in this region. The full-length cDNA obtained by the two-step reverse transcription was amplified successfully by long PCR and subsequently cloned into a plasmid vector. The cloned cDNA showed toxicity and proved to be unstable when amplified in E. coli. PMID- 10381088 TI - Improved detection of bovine herpesvirus 1 in artificially infected bovine semen by protein amplification. AB - Infection with bovine herpesvirus 1 (BHV 1) occurs worldwide and causes serious economic losses due to loss of animals, abortions, decreased milk production, and loss of body weight. There is a real need for sensitive diagnostic procedures for detection of the presence of virus in order to achieve effective control of BHV 1 induced diseases. BHV 1 is frequently found in bovine semen and can be widely transmitted through artificial insemination. Thus the detection of BHV 1 in artificial insemination centers and semen banks is of crucial importance in the control of its dissemination to the cattle industry, worldwide. In the present study, a protein amplification assay following polymerase chain reaction (PCR) of the highly conserved BHV 1 glycoprotein D gene was used in order to improve the sensitivity of direct virus detection in bovine semen. This method of BHV 1 detection is at least 200 orders of magnitude more sensitive than traditional PCR and would have direct clinical applications in antigen-based detection tests. In this method, amplification of the BHV 1 gD gene by PCR is followed by a coupled in vitro transcription translation of a small aliquot from the reaction. When the transcription translation was carried out in the presence of [35S]methionine and the products analyzed by SDS PAGE and autoradiography, 0.0014 TCID50 of virus could be detected in raw bovine semen in contrast to 0.28 TCID50 of virus detected using traditional PCR. Given the limitations in the method used for protein detection, this 'in vitro protein amplification' has the potential of attaining superior sensitivity for direct virus detection in clinical samples. PMID- 10381087 TI - Selection of phage-display peptides that bind to cucumber mosaic virus coat protein. AB - Several discrete peptides that bind specifically to the coat protein of cucumber mosaic virus (CMV) were isolated from a diverse phage library displaying random nonapeptides on the major coat protein VIII. Enrichment was shown by polyclonal phage enzyme linked immunosorbent assay (ELISA) after three rounds of selection. Sequencing of the genes encoding 10 of these peptides revealed an absence of any conserved motifs, although nine of them contained a high proportion of proline residues. Some of the selected peptides were displayed at the N-terminus of thioredoxin and expressed in the cytoplasm of Escherichia coli. Both the phage displayed and thioredoxin-fusion versions of the peptides could detect purified CMV and CMV present in crude leaf extracts from infected plants. By dot blot analysis, a thioredoxin-peptide fusion could readily detect as little as 5 ng of CMV. The peptides did not bind to other plant viruses. These peptides have been shown to be specific and highly sensitive tools in the detection of CMV and, as well as their diagnostic potential, they could form the basis for a novel disease resistance strategy. PMID- 10381089 TI - High level expression of hepatitis B virus surface antigen in stably transfected Drosophila Schneider-2 cells. AB - Two transfer vector systems have been constructed for the generation of Drosophila melanogaster Schneider-2 (DS-2) cells transfected stably and used to express the small surface antigen of hepatitis B virus (HBsAg). One system is based on the cotransfection of an expression vector for the S gene under the control of an inducible Drosophila metallothionein (Mtn) promotor and a resistance plasmid which carries a selectable marker dihydrofolate reductase (dhfr) gene under the control of a Drosophila actin 5C distal promoter. The second system is based on the transfection of a single plasmid, which includes both expression units. Both vector systems were suitable for the generation of stably transfected DS-2 cell-lines secreting high levels of HBsAg. The quantities of HBsAg expression from polyclonal DS-2 cells correlated strictly with the concentration of the transfected S gene expression vector. Clonal cell-lines selected from the most efficient HBsAg producing polyclonal cell-populations were examined in more detail. All of the transfected S genes were found to be integrated and the copy numbers per genome varied extremely between 10 and 240. Furthermore, the levels of secreted HBsAg varied greatly between different clones and in best they reached up to 7 microg/ml under serum-free cell culture conditions. Thus, DS-2 cells transfected stably provide an alternative source for the production of HBsAg particles for diagnostic purposes and vaccine development. PMID- 10381091 TI - Assay of humoral immunity to mumps virus. AB - One hundred randomly chosen sera from blood donors from North London were assayed for antibodies to mumps virus by plaque reduction and microtitre neutralisation assay, haemagglutination inhibition and in-house ELISA. The assay reproducibility was determined, and there was reasonable agreement between antibody levels measured by the two neutralising methods. Neutralising antibody levels measured by either method were low but the strain used in the assay had a large effect on the antibody titres observed. Titres measured by neutralisation assay, HI assay and ELISA did not correlate well. Assessment of immunity to mumps virus remains problematical. PMID- 10381090 TI - Purification and characterization of hepatitis B virus surface antigen particles produced in Drosophila Schneider-2 cells. AB - The small surface antigen of hepatitis B virus (HBV) was produced in Drosophila melanogaster Schneider-2 (DS-2) cells transfected stably using an inducible Drosophila metallothionein promoter. Selected clonal DS-2 cell-lines expressed and secreted large quantities of HBsAg particles consisting exclusively of non glycosylated 25 kDa proteins. HBsAg produced by DS-2 cells had physical and biochemical properties very similar to 22 nm particles derived from the human hepatoma cell-line PLC/PRF/5. DS-2 cell-derived HBsAg particles were purified near homogeneity by a strategy based on protein concentration, precipitation and ultracentrifugation. The resulting HBsAg product was < 98% pure. A single immunisation of BALB/c mice with both DS-2 and yeast-cell derived purified HBsAg particles without adjuvants elicited a substantial humoral antibody and class-I restricted cytotoxic T lymphocyte (CTL) response. Adsorption of HBsAg particles to aluminium hydroxide resulted in increased levels of HBsAg-specific antibodies. However, CTLs were not elicited by HBsAg/Alum combinations. Thus, stably transfected DS-2 cells provide a useful source for the production of HBV subviral particles for diagnostic and research purposes as well as for novel vaccine development. PMID- 10381092 TI - Quantification of hepatitis C virus RNA in serum by branched DNA-based signal amplification assays. AB - The objective of the study was to compare the clinical sensitivity and specificity of versions 1.0 and 2.0 of the branched DNA (bDNA)-based hepatitis C virus (HCV) RNA quantification assay, and also to compare the values yielded by the two versions according to the HCV genotype. Serum samples from 268 patients tested routinely by a non-quantitative HCV RNA PCR assay (group A) were tested with version 2.0 of the bDNA assay. Samples from 342 HCV PCR-positive patients with chronic hepatitis C eligible for interferon treatment (group B) were tested with both version 1.0 and version 2.0 of the bDNA assay. Version 2.0 had a clinical sensitivity of 92% (95% confidence interval (CI): 87-97%) in group A and 89% (86-92%) in group B. In group B, the gain in sensitivity with bDNA 2.0 was 16% relative to bDNA 1.0 (P < 0.001). The log values of the two assays correlated with samples positive by both assays (r = 0.83, P < 0.0001), but the distribution of values was larger in samples containing HCV genotypes 2 and 3. The mean ratio of assay 2.0/assay 1.0 values was 1.69 +/- 1.44 (range: 0.33-13.43). The mean ratio was close to 1 with samples containing genotype 1 or 4, but ranged from 0.33 to more than 5. The mean ratio was close to 3 with samples containing genotype 2 or 3, and ranged from 0.5 to more than 13. HCV RNA levels were significantly lower in samples containing genotype 4 than in those containing other genotypes. Sera from 200 anti-HCV-negative, HCV RNA PCR-negative blood donors (group C), and from 164 anti-HCV-negative patients with symptoms of chronic liver disease (group D) were used to assess the clinical specificity of bDNA 2.0. In addition, samples with an HCV RNA titer between 0.2 (assay cutoff) and 0.5 MEq/ml from a group of 546 patients tested routinely for HCV RNA load by bDNA 2.0 (group E) were retested by bDNA 2.0 and by qualitative PCR. The specificity of bDNA 2.0 was 100% (98-100%) in group C and 99% (97-100%) in group D. Among the 41 samples from group E, 38 were positive by bDNA 2.0 retesting (36 were PCR-positive) and three were negative by bDNA 2.0 retesting (all were PCR positive). It is concluded that version 2.0 of the bDNA assay is markedly more sensitive than version 1.0 and has a good specificity. In contrast with version 1.0, version 2.0 is not influenced by the HCV genotype. The relationship between values obtained with assays 1.0 and 2.0 on clinical specimens is not linear, indicating that HCV RNA titers cannot reliably be calculated from the results of version 1.0. PMID- 10381093 TI - Molecular characterization of Brazilian avian pneumovirus isolates: comparison between immunochemiluminescent Southern blot and nested PCR. AB - Avian pneumovirus (APV) causes acute respiratory tract infection both in turkeys (turkey rhinotracheitis) and chickens (swollen head syndrome (SHS)) with sudden onset and rapid spread through the flocks. In this study, an immunochemiluminescent Southern blot RT-PCR assay was employed to detect a F gene transcript of the APV in two European turkey isolates and two Brazilian chicken isolates. Limiting dilution PCR was carried out to compare the sensitivity of immunochemiluminescent Southern blot assay and nested PCR assay (nPCR). The sensitivity and specificity of immunochemiluminescent Southern blot RT-PCR assay were comparable to that of nPCR, and at least 100 fold more sensitive than a single PCR amplification. Sequence analysis of the 175 bp product of the F gene revealed 100% identity with APV sequences described earlier. PMID- 10381094 TI - Integrons and gene cassettes: a genetic construction kit for bacteria. PMID- 10381096 TI - The distribution of mecA, mecR1 and mecI and sequence analysis of mecI and the mec promoter region in staphylococci expressing resistance to methicillin. AB - The presence and sequences of genes that regulate the expression of methicillin resistance was investigated in 42 isolates of Staphylococcus aureus and 102 isolates of coagulase-negative staphylococci (CNS). PCR was used to detect mecA and the regulatory genes mecR1 and mecI. In a selected group of isolates, the sequences of mecI and the mec promoter region were also determined and compared with the sequences obtained from pre-MRSA strain N315. The genetic diversity of the collection was assessed by pulsed-field gel electrophoresis (PFGE). mecA was present in 21 S. aureus and 44 CNS. mecR1 was associated with mecA in all S. aureus and in all CNS, except two isolates of Staphylococcus haemolyticus. mecI was present in 48% of mecA-positive S. aureus and 50% of mecA-positive CNS. In six S. aureus isolates, mecI contained a termination codon at nucleotide 202 which would truncate the MecI protein. No mutation was found in the mecI gene of the four other S. aureus and 15 CNS sequenced. Seven isolates of Staphylococcus simulans had a single nucleotide substitution in the mec promoter region. Expression of methicillin resistance could be explained for all mecA-positive staphylococci with mutations within mecI or in the mec promoter region or in which mecI was deleted. However, the 'wild type' sequences observed in four S. aureus and eight CNS suggest that there is another mechanism for overcoming the repression of resistance caused by mecI. PMID- 10381095 TI - Host range of the ermF rRNA methylase gene in bacteria of human and animal origin. AB - We screened 183 different clinical anaerobic and aerobic bacteria isolated from humans and other animals for the presence of the ermF gene using a polymerase chain reaction (PCR) assay. The ermF gene was detected in 107 (58%) clinical isolates, including 42 (61%) of 69 gram-positive bacteria and 65 (57%) of 114 gram-negative bacteria. Twenty-five ATCC isolates were also tested; 20 (80%) carried the ermF gene. The gene products from the ermF PCR from four isolates were sequenced and showed 95-99% nucleotide homology with the ermF gene and 98 99% amino acid homology with the gene product. Eleven (58%) of the 19 gram negative donors tested were able to transfer the ermF gene. All nine (100%) of the gram-positive donors tested transferred the ermF gene, using either Enterococcus faecalis or Haemophilus influenzae as the recipients. PMID- 10381097 TI - Aspartic acid for asparagine substitution at position 276 reduces susceptibility to mechanism-based inhibitors in SHV-1 and SHV-5 beta-lactamases. AB - In SHV-type beta-actamases, position 276 (in Ambler's numbering scheme) is occupied by an asparagine (Asn) residue. The effect on SHV-1 beta-lactamase and its extended-spectrum derivative SHV-5 of substituting an aspartic acid (Asp) residue for Asn276 was studied. Mutations were introduced by a PCR-based site directed mutagenesis procedure. Wild-type SHV-1 and -5 beta-lactamases and their respective Asn276-->Asp mutants were expressed under isogenic conditions by cloning the respective bla genes into the pBCSK(+) plasmid and transforming Escherichia coli DH5alpha. Determination of IC50 showed that SHV-1(Asn276-->Asp), compared with SHV-1, was inhibited by 8- and 8.8-fold higher concentrations of clavulanate and tazobactam respectively. Replacement of Asn276 by Asp in SHV-5 beta-lactamase caused a ten-fold increase in the IC50 of clavulanate; the increases in the IC50s of tazobactam and sulbactam were 10- and 5.5-fold, respectively. Beta-lactam susceptibility testing showed that both Asn276-->Asp mutant enzymes, compared with the parental beta-lactamases, conferred slightly lower levels of resistance to penicillins (amoxycillin, ticarcillin and piperacillin), cephalosporins (cephalothin and cefprozil) and some of the expanded-spectrum oxyimino beta-lactams tested (cefotaxime, ceftriaxone and aztreonam). The MICs of ceftazidime remained unaltered, while those of cefepime and cefpirome were slightly elevated in the clones producing the mutant beta lactamases. The latter clones were also less susceptible to penicillin-inhibitor combinations. Asn276-->Asp mutation was associated with changes in the substrate profiles of SHV-1 and SHV-5 enzymes. Based on the structure of TEM-1 beta lactamase, the potential effects of the introduced mutation on SHV-1 and SHV-5 are discussed. PMID- 10381098 TI - In-vitro activity of 29 antimicrobial agents against penicillin-resistant and intermediate isolates of Streptococcus pneumoniae. AB - Antibiotic resistance among isolates of Streptococcus pneumoniae is increasing worldwide. Optimal therapy, though unknown, should be guided by in-vitro susceptibility testing. Currently, vancomycin is the only approved antibiotic that is universally active against multiresistant S. pneumoniae. In-vitro activities were determined for 29 antimicrobial agents against 22 penicillin intermediate S. pneumoniae (PISP) and 16 penicillin-resistant S. pneumoniae (PRSP) isolates. MICs were determined in cation-adjusted Mueller-Hinton broth with 3% lysed horse blood in microtitre trays. Antimicrobial classes tested included cephalosporins, penicillin, aminopenicillins, macrolides, quinolones, carbapenems and other antimicrobial agents. Among the classes of antimicrobial agents tested, wide differences in susceptibility were demonstrated for both PISP and PRSP. Of the cephalosporins, ceftriaxone and cefotaxime demonstrated the best in-vitro activity for both PISP and PRSP. Of the quinolones, clinafloxacin and trovafloxacin showed the greatest in-vitro activity. Rifampicin and teicoplanin demonstrated excellent in-vitro activity. Promising in-vitro results of newer agents, such as quinupristin/dalfopristin, ramoplanin, teicoplanin and linezolid may justify further evaluation of these agents in clinical trials. PMID- 10381099 TI - An in-vitro study of the antimicrobial susceptibilities of Yersinia enterocolitica and the definition of a database. AB - The antibiotic susceptibilities of 151 Yersinia enterocolitica strains isolated from humans, animals and the environment, and belonging to biovars 1A (n = 22), 1B (n = 13), 2 (n = 12), 3 (n = 31), 4 (n = 63) and 5 (n = 10) to 71 antibiotics were examined. Biovars could be characterized phenotypically by their similar antibiograms. For the majority of biovars only minor differences in susceptibilities were detected. Ninety-nine percent of all Yersinia strains were resistant to amoxycillin, but biovar-related differences in MICs were demonstrated. Significant differences, characteristic of specific biovars, were detected for fosfomycin, some beta-lactams (in particular amoxycillin/clavulanate, and ticarcillin). Strains of biovar 1A were resistant or of intermediate susceptibility to ticarcillin and had fosfomycin MICs of 16-64 mg/L and amoxycillin/clavulanate MICs of 4-32 mg/L. Most strains of biovar 1B (n = 11) were sensitive to amoxycillin/clavulanate (MIC = 0.5-2 mg/L). Most strains of biovar 2 (n = 10) showed an antibiotic phenotype resembling that of strains of biovar 1A, but they were more susceptible to fosfomycin (MIC = 4-16 mg/L). Twenty seven strains of biovar 3 had amoxycillin/clavulanate MICs of 4-32 mg/L; 19 of them were sensitive to ticarcillin and fosfomycin. Biovar 4 strains were uniformly sensitive or of intermediate susceptibility to amoxycillin/clavulanate (MIC = 0.5-4 mg/L), sensitive to fosfomycin (MIC = 1-4 mg/L) but resistant to ticarcillin. The prevailing antibiotic phenotype of biovar 5 (n = 7) was similiar to the phenotype of biovar 4, although these biovar 5 strains were slightly more susceptible to amoxycillin/clavulanate and showed a heterogeneous pattern of susceptibility to fosfomycin (MIC = 1-32 mg/L). The results of this study show that unambiguous statements cannot be made about the natural antibiotic susceptibility to certain beta-lactams and fosfomycin of Y. enterocolitica. The data indicate a complex regulation of beta-lactamases in Y. enterocolitica. Some beta-lactamases are found more frequently or will be expressed predominantly in specific biovars. PMID- 10381100 TI - Amoxycillin tolerance in Helicobacter pylori. AB - Resistance to amoxycillin in Helicobacter pylori has only recently been reported. To demonstrate the existence of resistance, and to test for the presence of tolerance, 17 amoxycillin-resistant strains of H. pylori, first isolated in Sardinia (Italy) and the USA, were studied. Four amoxycillin-sensitive strains were used as controls. Primary isolates of all test strains exhibited amoxycillin resistance; beta-lactamase activity was not detected. Amoxycillin resistance was lost after storage of strains at -80 degrees C but could be rescued by plating these strains on to amoxycillin gradient plates. MICs and MBCs from rescued isolates ranged from 0.5 to 32 mg/L and from 32 to > 1024 mg/L, respectively. MBC/MIC ratios > or = 32 are characteristic of antibiotic tolerance. The ratios of MBC/MIC of amoxycillin ranged from 32 to > 1024 for the test strains, indicating that these strains were tolerant to the antibiotic. Amoxycillin resistance does occur in H. pylori. Amoxycillin susceptibility testing of H. pylori isolates in patients who fail therapy should include determination of the MBC to detect tolerance. PMID- 10381101 TI - Markedly different rates and resistance profiles exhibited by seven commonly used and newer beta-lactams on the selection of resistant variants of Enterobacter cloacae. AB - Seven beta-lactam antibiotics (cefepime, cefoperazone, ceftazidime, ceftriaxone, cefamandole, imipenem and meropenem) were tested for their potential to select resistance in standard and clinical strains of Enterobacter cloacae (n = 9). The strains were subcultured daily with the test antibiotics at doubling concentrations starting at 0.125 x MIC. Development of resistance throughout the passages was detected by a disc diffusion test. Ceftazidime, ceftriaxone and cefamandole selected resistance at a faster rate than cefoperazone, cefepime and meropenem. Imipenem did not select resistance in the nine strains tested and was the only antibiotic that eradicated all the strains during selection. The resistance patterns of strains selected by meropenem, cefepime and the other cephalosporins were markedly different, although cross-resistance to the early generation cephalosporins was common. The resistance phenotypes of most strains remained stable upon serial passages in antibiotic-free medium. The findings of this study highlight the importance of the choice of antibiotic for therapy not only on the basis of its antibacterial activity, but also on its potential to select resistance to itself and other antibiotics. PMID- 10381102 TI - Quinolone accumulation by Pseudomonas aeruginosa, Staphylococcus aureus and Escherichia coli. AB - The accumulation of nalidixic acid and 14 fluoroquinolones over a range of external drug concentrations (10-100 mg/L; c. 25-231 microM) into intact cells of Escherichia coli KL-16, Staphylococcus aureus NCTC 8532, Pseudomonas aeruginosa NCTC 10662 and spheroplasts of E. coli was investigated. The effect of 100 microM carbonyl cyanide m-chlorophenyl hydrazone (CCCP) upon the concentration of quinolone accumulated by intact cells and spheroplasts of E. coli was also determined. Except for pefloxacin, there was an increase in the concentration of the six quinolones examined accumulated by E. coli, despite a reduction in fluorescence at alkaline pH. For ciprofloxacin the partition coefficient (P(app)) was constant despite an increase in the pH; however, the P(app) for nalidixic acid decreased significantly with an increase in pH. The concentration of nalidixic acid, ciprofloxacin and enrofloxacin accumulated by E. coli and S. aureus increased with an increase in temperature up to 40 degrees C and 50 degrees C, respectively. Above these temperatures the cell viability decreased. With an increase in drug concentration there was, for intact E. coli and 12/15 agents, and for S. aureus and 10/15 agents, a linear increase in the concentration of drug accumulated. However, for P. aeruginosa and 13/15 agents there was apparent saturation of an accumulation pathway. Assuming 100% accumulation into intact cells of E. coli, for 10/14 fluoroquinolones < or = 40% was accumulated by spheroplasts. CCCP increased the concentration of quinolone accumulated but the increase varied with the agent and the bacterial species. The variation in the effect of CCCP upon accumulation of the different quinolones into E. coli could result from chemical interactions or from different affinities of the proposed efflux transporter for each quinolone. Overall, these data suggest that accumulation of most quinolones into E. coli and S. aureus proceeds by simple diffusion, but that P. aeruginosa behaves differently. PMID- 10381103 TI - The post-exposure response of Enterobacteriaceae to ceftibuten. AB - The responses of ten isolates of Enterobacteriaceae to ceftibuten exposure were monitored by measuring several parameters. Post-antibiotic effect (PAE), control related effective regrowth time (CERT) and post-antibiotic sub-MIC effect (PA SME) were determined by bacterial enumeration carried out either by impedance in combination with viable counting (IMP/VC) or by impedance in combination with bioluminescence (IMP/BIOL). Kill curves were carried out by bioluminescence, viable counting and direct microscopy and post-exposure morphology was established. Ceftibuten primarily provoked filamentation. Over 24 h, kill of up to 3.6 log10 was evident by viable counting and direct microscopy at and above the MIC. Minimal kill, of up to 0.26 log10, was shown by bioluminescence. PAE was found to be method dependent, with statistical differences established by Student's t-test. PAE values of up to 0.48 h and 1.47 h (by IMP/BIOL and IMP/VC respectively) were not concentration dependent above 1 x MIC. CERT values were not method dependent, with values of up to 1.71 h also showing a lack of concentration dependence above 1 x MIC. PA-SME may reflect the situation in vivo more accurately than either PAE or CERT. In PAE and CERT studies the antibiotic is eliminated almost immediately, whereas in vivo there is gradual decrease in antibiotic levels. These persisting levels are reflected more accurately by PA SME. Compared with PAE and CERT, significantly longer values, of up to 7.27 h, were obtained by PA-SME, although this parameter was also found to be method dependent. The results of the PA-SME studies, which may be the most clinically relevant pharmacodynamic parameter, confirm the appropriateness of the current once- or twice-daily dosing schedules despite the lack of PAE. PMID- 10381104 TI - Pharmacodynamics of levofloxacin and ciprofloxacin against Streptococcus pneumoniae. AB - An in-vitro pharmacokinetic model was used to compare the pharmacodynamics of levofloxacin and ciprofloxacin against four penicillin-susceptible and four penicillin-resistant Streptococcus pneumoniae. Logarithmic-phase cultures were exposed to the peak concentrations of levofloxacin or ciprofloxacin observed in human serum after 500 mg and 750 mg oral doses, human elimination pharmacokinetics were simulated, and viable bacterial counts were measured at 0, 1, 2, 4, 6, 8, 12, 24 and 36 h. Levofloxacin was rapidly and significantly bactericidal against all eight strains evaluated, with eradication of six strains occurring despite area under the inhibitory curve over 24 h (AUIC24) values of only 32-64 SIT(-1) x h (serum inhibitory titre over time). The pharmacodynamics of ciprofloxacin were more variable and the rate of bacterial killing was consistently slower than observed with levofloxacin. Ciprofloxacin eradicated five strains despite having an AUIC24 of only 44 SIT(-1) x h. These data suggest that the increased potency of levofloxacin and more favourable pharmacokinetics compared with ciprofloxacin provide enhanced pharmacodynamic activity against S. pneumoniae. Furthermore, these data suggest that the minimum AUIC required for clinical efficacy against and eradication of S. pneumoniae with levofloxacin and ciprofloxacin may be well below the 125 SIT(-1) x h identified by other studies. PMID- 10381105 TI - Quinupristin/dalfopristin attenuates the inflammatory response and reduces the concentration of neuron-specific enolase in the cerebrospinal fluid of rabbits with experimental Streptococcus pneumoniae meningitis. AB - The inflammatory response following initiation of antibiotic therapy and parameters of neuronal damage were compared during intravenous treatment with quinupristin/dalfopristin (100 mg/kg as either a short or a continuous infusion) and ceftriaxone (10 mg/kg/h) in a rabbit model of Streptococcus pneumoniae meningitis. With both modes of administration, quinupristin/dalfopristin was less bactericidal than ceftriaxone. However, the concentration of proinflammatory cell wall components (lipoteichoic acid (LTA) and teichoic acid (TA)) and the activity of tumour necrosis factor (TNF) in cerebrospinal fluid (CSF) were significantly lower in the two quinupristin/dalfopristin groups than in ceftriaxone-treated rabbits. The median LTA/TA concentrations (25th/75th percentiles) were as follows: (i) 14 h after infection: 133 (72/155) ng/mL for continuous infusion of quinupristin/dalfopristin and 193 (91/308) ng/mL for short duration infusion, compared with 455 (274/2042) ng/mL for ceftriaxone (P = 0.002 and 0.02 respectively); (ii) 17 h after infection: 116 (60/368) ng/mL for continuous infusion of quinupristin/dalfopristin and 117 (41/247) ng/mL for short duration infusion, compared with 694 (156/2173) ng/mL for ceftriaxone (P = 0.04 and 0.03 respectively). Fourteen hours after infection the median TNF activity (25th/75th percentiles) was 0.2 (0.1/1.9) U/mL for continuous infusion of quinupristin/dalfopristin and 0.1 (0.01/3.5) U/mL for short duration infusion, compared with 30 (4.6/180) U/mL for ceftriaxone (P = 0.02 for each comparison); 17 h after infection the TNF activity was 2.8 (0.2/11) U/mL (continuous infusion of quinupristin/dalfopristin) and 0.1 (0.04/6.1) U/mL (short duration infusion), compared with 48.6 (18/169) U/mL for ceftriaxone (P = 0.002 and 0.001). The concentration of neuron-specific enolase (NSE) 24 h after infection was significantly lower in animals treated with quinupristin/dalfopristin: 4.6 (3.3/5.7) microg/L (continuous infusion) and 3.6 (2.9/4.7) microg/L (short duration infusion) than in those treated with ceftriaxone (17.7 (8.8/78.2) microg/L) (P = 0.03 and 0.009 respectively). In conclusion, antibiotic treatment with quinupristin/dalfopristin attenuated the inflammatory response within the subarachnoid space after initiation of antibiotic therapy. The concentration of NSE in the CSF, taken as a measure of neuronal damage, was lower in quinupristin/dalfopristin-treated rabbits than in ceftriaxone-treated rabbits. PMID- 10381106 TI - Liposomal nystatin against experimental pulmonary aspergillosis in persistently neutropenic rabbits: efficacy, safety and non-compartmental pharmacokinetics. AB - The activity of liposomal nystatin against invasive pulmonary aspergillosis was investigated in persistently neutropenic rabbits. Treatment groups included liposomal nystatin at dosages of 1, 2 and 4 mg/kg/day intravenously, or amphotericin B deoxycholate 1 mg/kg/day administered intravenously after normal saline loading. As compared with untreated controls, liposomal nystatin administered at 2 and 4 mg/kg/day prolonged survival and reduced fungus-mediated tissue injury and excess lung weight at post-mortem in a similar manner to amphotericin B. Although amphotericin B was superior in clearing infected lung tissue, treatment with all regimens of liposomal nystatin led to a significant reduction in pulmonary fungal tissue burden. During treatment, ultrafast CT-scan demonstrated ongoing resolution of pulmonary lesions at 2 and 4 mg/kg/day, but not at 1 mg/kg/day. With the exception of mild increases in blood urea nitrogen (BUN) and serum creatinine values during treatment at 2 and 4 mg/kg/day, which were similar to those found in amphotericin B-treated rabbits, liposomal nystatin was well tolerated. Preliminary pharmacokinetic studies in non-infected animals established linear drug disposition of liposomal nystatin in plasma over the investigated dosage range and peak plasma levels above the MIC for the test strain after multiple daily dosing for 7 days. Liposomal nystatin increased survival and provided reduced tissue injury, effective microbiological clearance and tolerable side effects in experimental pulmonary aspergillosis in persistently neutropenic rabbits, thus providing a rational basis for further investigations in clinical trials. PMID- 10381108 TI - The clinical efficacy of continuous-infusion flucloxacillin in serious staphylococcal sepsis. AB - Since the efficacy of beta-lactams against pathogens such as methicillin susceptible Staphylococcus aureus (MSSA) is related to the time for which serum drug concentrations exceed the MIC for the pathogen, administration of anti staphylococcal beta-lactams by continuous infusion may provide a more suitable means of drug delivery than intermittent dosing. To assess the clinical efficacy of continuous-infusion therapy, we reviewed the outcomes for 20 consecutive patients with proven serious MSSA sepsis (three with endocarditis, ten osteomyelitis, one endocarditis plus osteomyelitis and six deep abscess) treated with continuous-infusion flucloxacillin (8-12 g/day). Patients initially receiving routine intermittent-dose flucloxacillin therapy were changed to continuous-infusion flucloxacillin (mean duration 29 days; range 4-60 days) for completion of their treatment course. In the majority of cases this was given at home. Serum flucloxacillin concentrations during continuous-infusion flucloxacillin 12 g/day were 11.5->40 mg/L (ten patients) and those during continuous-infusion flucloxacillin 8 g/day were 8->40 mg/L (five patients), these concentrations being well above the expected MIC of flucloxacillin for MSSA. Continuous-infusion flucloxacillin was well tolerated by most patients, and 14/17 patients (82%) who completed their course of continuous-infusion flucloxacillin were judged clinically and microbiologically cured at long-term follow-up (mean 67 weeks; range 4-152 weeks). These preliminary data suggest that, following initial intermittent-dose flucloxacillin therapy, continuous-infusion flucloxacillin is an effective treatment option for serious MSSA sepsis, and forms a feasible and possibly preferable alternative to glycopeptides when considering home-based parenteral therapy for these infections. Further studies are needed to identify whether continuous-infusion flucloxacillin can entirely replace intermittent-dose therapy for such infections. PMID- 10381107 TI - Cost-effectiveness of prophylactic nasal mupirocin in patients undergoing peritoneal dialysis based on a randomized, placebo-controlled trial. AB - The study objective was to measure the benefits of elimination of nasal carriage of Staphylococcus aureus by calcium mupirocin ointment in patients undergoing continuous ambulatory peritoneal dialysis. The design was a prospective, placebo controlled, randomized clinical trial. The subjects were 267 patients recruited from nine renal units in Belgium, France and the UK. The main outcome measures were the rate of catheter exit site infection (ESI), rates of other infections and healthcare costs from the perspective of a hospital budget-holder. The rate of ESI caused by S. aureus was significantly reduced from one in 28.1 patient months to one in 99.3 patient months (P = 0.006) and there were also non significant trends towards lower rates of ESI caused by any organism and peritonitis caused by S. aureus. In comparison with the placebo group, patients in the mupirocin group with ESI had lower antibiotic (P = 0.02) and hospitalization costs (P = 0.065). However, overall costs of antibiotic treatment, for all infections combined, were not significantly different (P = 0.2) and total antibiotic costs (including mupirocin) were significantly higher in the mupirocin group (P = 0.001). Mupirocin prophylaxis would have been cost neutral if the rate of ESI increased to >75% in the placebo group, or if all healthcare costs increased by 40%, or if the cost of screening was reduced from Pound Sterling 15 to Pound Sterling 3 per patient, or if the cost of mupirocin treatment was reduced from Pound Sterling 93 to Pound Sterling 40 per patient year. In conclusion, savings in healthcare costs are unlikely to be sufficiently great to offset the cost of mupirocin and screening for nasal carriage of S. aureus. The decision about whether or not to implement mupirocin should depend on a local analysis of the value of preventing ESIs caused by S. aureus. PMID- 10381109 TI - Risk of resistance related to antibiotic use before admission in patients with community-acquired bacteraemia. AB - We analysed the association of antibiotic therapy before admission and antibiotic resistance of blood isolates in a total of 1717 community-acquired bacteraemias in the County of Northern Jutland during 1992-96. Antibiotics had been prescribed to 14% of the patients during the 30 days before admission and to 37% during the 6 months. The most frequently prescribed antibiotics within 30 days were ampicillin (28%), penicillin G (27%), sulphonamides and/or trimethoprim (16%) and macrolides (14%). The most frequent blood isolates were Escherichia coli (33%), other Enterobacteriaceae (8%), Streptococcus pneumoniae (23%) and Staphylococcus aureus (10%). Of the 575 isolates of E. coli, 425 (74%), 432 (75%) and 518 (90%) were susceptible to ampicillin, sulphonamides and trimethoprim, respectively. Previous antibiotic prescriptions were strongly associated with resistance to ampicillin, sulphonamides and trimethoprim in E. coli. The association was less pronounced for S. aureus and enteric rods other than E. coli. Antibiotic prescriptions within the last 3 months predicted antibiotic resistance, and this should be taken into account when selecting empirical antibiotic therapy of severe community-acquired infections. PMID- 10381110 TI - Biochemical characteristics of a carbapenemase from an Acinetobacter baumannii isolate collected in Buenos Aires, Argentina. AB - Three carbapenem-resistant Acinetobacter baumannii isolates were collected at a hospital in Buenos Aires, Argentina. Isoelectric focusing revealed multiple beta lactamases, with two of the isolates showing identical profiles. A pI 6.9 carbapenemase with a molecular weight of 30 kDa was purified from one of these two isolates. The enzyme was predominantly a penicillinase, with its highest Vmax for oxacillin but highest Vmax/Km for benzylpenicillin. First-generation cephalosporins and imipenem were weaker substrates than penicillins, and oxyimino aminothiazolyl cephalosporins were essentially stable. Meropenem-hydrolysing activity was not detected, despite resistance. The carbapenemase was inhibited by clavulanic acid and tazobactam, but not by EDTA. These kinetics place the enzyme into functional group 2; as an oxacillinase it could be placed in sub-group 2d or, as a zinc-independent carbapenemase, in sub-group 2f. PMID- 10381112 TI - Fosfomycin tromethamine susceptibility of outpatient urine isolates of Escherichia coli and Enterococcus faecalis from ten North American medical centres by three methods. AB - One hundred percent of 1097 Escherichia coli and 97.5% of 157 Enterococcus faecalis isolates from outpatient urine specimens at ten North American medical centres were susceptible to fosfomycin tromethamine. The Etest MICs correlated well with those of agar dilution. Disc diffusion zone diameters correlated well with MICs and supported the previously proposed zone diameter breakpoints for fosfomycin. PMID- 10381111 TI - In-vitro susceptibilities of species of the Bacteroides fragilis group to newer beta-lactam agents. AB - The in-vitro activities of imipenem and four beta-lactam-beta-lactamase inhibitor combinations were tested against 816 strains of the Bacteroides fragilis group, and compared with other anti-anaerobic agents. None of the strains was resistant to metronidazole, and only one was resistant to chloramphenicol. Mezlocillin and piperacillin were moderately active, while clindamycin was the least active. Rates of resistance varied between various species. The new beta-lactam agents tested showed excellent activity; piperacillin-tazobactam and imipenem were the most active. The emergence of strains that are resistant to these agents, observed in this study, suggests there is a need to perform periodic antimicrobial susceptibility tests. PMID- 10381113 TI - Study to assess the reliability of a disc diffusion method for determining the sensitivity of gram-positive pathogens to dalfopristin/quinupristin. AB - A standardized method of disc testing the sensitivity of gram-positive pathogens to dalfopristin/quinupristin was developed, and then 'field tested' in ten centres in the UK. For a 15 microg disc, zone diameter breakpoints of 20 mm and 15 mm are suggested when organisms are tested on Iso-Sensitest agar and Iso Sensitest agar supplemented with 5% whole horse blood, respectively. PMID- 10381114 TI - Increasing fluoroquinolone resistance in salmonella serotypes in Finland during 1995-1997. AB - Antimicrobial resistance trends were examined for 811 salmonella isolates from humans collected in Finland during 1995-1997. The material was divided into domestic and foreign isolates according to the origin of infection. A total of 2.3% of the 387 domestic and 7.8% of the 424 foreign isolates were quinolone resistant (P < 0.001). Among the domestic isolates we detected an emergence of ciprofloxacin resistance (MIC > or = 0.25 mg/L) with the proportion of resistant isolates increasing from 0 to 2.2% (P = 0.2). Among the foreign isolates this increase was even more dramatic, from 2.0% to 8.4% (P = 0.037) during the study period. PMID- 10381115 TI - Comparative in-vitro activity of voriconazole (UK-109,496) and six other antifungal agents against clinical isolates of Scedosporium prolificans and Scedosporium apiospermum. AB - We report the in-vitro susceptibility of 27 clinical isolates of Scedosporium apiospermum and 43 of Scedosporium prolificans. S. apiospermum was resistant to fluconazole and flucytosine, with variable susceptibility to amphotericin B, itraconazole, ketoconazole and susceptible to miconazole. Voriconazole was much more active than fluconazole and flucytosine, more active than amphotericin B, itraconazole and ketoconazole and was as active as miconazole against S. apiospermum isolates. Voriconazole and the other six antifungal agents showed low activity against S. prolificans isolates. PMID- 10381116 TI - The concentrations of clinafloxacin in alveolar macrophages, epithelial lining fluid, bronchial mucosa and serum after administration of single 200 mg oral doses to patients undergoing fibre-optic bronchoscopy. AB - The concentrations of clinafloxacin were measured in serum, bronchial mucosa, alveolar macrophages and epithelial lining fluid after single 200 mg oral doses of clinafloxacin had been administered to 15 subjects who were undergoing bronchoscopy. Concentrations were measured using a microbiological assay method. Mean concentrations in serum, bronchial mucosa, alveolar macrophages and epithelial lining fluid at a mean of 1.27 h post-dose were 1.54, 2.65, 15.60 and 2.71 mg/L respectively. These site concentrations exceeded the MIC90 for common respiratory pathogens and indicate that clinafloxacin is likely to be effective in the treatment of a wide range of respiratory tract infections. PMID- 10381117 TI - Extended-spectrum beta-lactamase-producing strain of Acinetobacter baumannii isolated from a patient in France. PMID- 10381118 TI - Adherent bacteria and activity of antibiotics. PMID- 10381119 TI - Increasing prevalence of methicillin resistance amongst Staphylococcus aureus blood culture isolates. PMID- 10381120 TI - Correlation between serotype and in-vitro antibiotic susceptibility of pneumococci isolated in Saudi Arabia. PMID- 10381121 TI - Classification of Streptococcus pneumoniae based on in-vitro susceptibility to oxyiminocephalosporins. PMID- 10381122 TI - General stress response master regulator rpoS is expressed in human infection: a possible role in chronicity. PMID- 10381123 TI - Dextran sulfate enhances the level of an oxidative DNA damage biomarker, 8-oxo 7,8-dihydro-2'-deoxyguanosine, in rat colonic mucosa. AB - Dextran sodium sulfate (DSS) given in drinking water can induce colonic inflammation and produce colorectal tumors in rodents, although it is not directly genotoxic. The hypothesis that DSS can produce free radicals and induce oxidative DNA damage in colonic mucosa has been tested. In rats fed for 2 days with water containing 3% and 6% DSS, colonic inflammation manifestations were recorded and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo), a major biomarker of oxidative DNA damage, was assayed in colonic mucosa. As compared with control rats given pure water, inflammatory manifestations were seen in rats given DSS. At the same time, 8-oxodGuo levels in colonic mucosa were doubled (P < 0.001). These results suggest that formation of oxidative DNA damage in colonic mucosa depends on inflammation and maybe on the production of reactive oxygen species. This study shows that DSS can induce oxidative DNA damage within only 2 days, which could explain in part its carcinogenic properties. PMID- 10381124 TI - The prognostic significance of tumor angiogenesis in Taiwanese patients with invasive ductal breast carcinomas. AB - Angiogenesis plays an important role in tumor growth and metastasis. We performed an immunohistochemical study by anti-CD31 antibody to investigate the prognostic significance of microvessel count (MVC) in 163 Taiwanese patients with primary invasive ductal breast carcinomas. Univariate analysis revealed that MVC per 200x field correlated significantly with disease-free (P < 0.001) and overall survival (P < 0.0008). In 74 patients with node-negative tumors, MVC also significantly correlated with disease-free (P < 0.03) and overall survival (P < 0.007). Multivariate Cox regression analysis demonstrated that MVC is an independent factor in predicting disease-free (P < 0.02) and overall survival (P < 0.02). Our study confirms that tumor angiogenesis as measured by MVC is a reliable independent prognostic factor of breast cancer. PMID- 10381125 TI - Expressions of vascular endothelial growth factor (VEGF) and its mRNA in uterine endometrial cancers. AB - To know the potential of growth, invasion and metastasis of uterine endometrial cancer associated with neovascularization, the expressions of VEGF and its mRNA, especially their subtypes, in uterine endometrial cancers and normal uterine endometria as controls were determined by Western blot analyses with a sandwich enzyme immunoassay and RT-PCR-Southern blot analysis, respectively, and the relation between their expressions and histological grades, grades of myometrial invasion and clinical stages of uterine endometrial cancers was analyzed. The levels of VEGF (VEGF165 and VEGF121) protein and mRNA were in a wide range and higher in normal uterine endometria than in the malignant counterparts. The levels of VEGF protein were higher in order of histopathological differentiation (normal uterine endometrium > well-differentiated (G1) > moderately differentiated (G2) and poorly differentiated (G3)) and those of VEGF protein and VEGF121 mRNA were lower in order of the advance of clinical stages (normal uterine endometrium > stage I > stage II > stages III and IV). There was, however, no significant difference in their levels among uterine endometrial cancers classified according to grades of myometrial invasion. This suggests that VEGF is downregulated during uterine endometrial cancer progression with dedifferentiation. Namely, VEGF in some endometrial cancers might contribute to the early process of advancing of malignancy via angiogenic activity. PMID- 10381126 TI - Absence of APC gene mutation in the mutation cluster region in hepatocellular carcinoma. AB - APC gene mutations have been demonstrated not only in colorectal carcinoma but also in a variety of human cancers. To define the possible role of mutations of the APC gene in hepatocarcinogenesis, we examined 46 pairs of hepatocellular carcinomas and corresponding non-tumorous liver tissue by polymerase chain reaction and single strand conformation polymorphism. All 46 hepatocellular carcinomas had no altered electrophoretic mobility to suggest the presence of APC gene mutation in the mutation cluster region. We also examined the possible loss of heterozygosity of APC and MCC gene loci by fragment length polymorphism analysis and by polymerase chain reaction. None of the cases showed a loss of heterozygosity at the APC and MCC gene loci. The results suggested that the possibility of APC and MCC as the gene defect in the genesis of human hepatocellular carcinoma may be very rare. PMID- 10381127 TI - Promotion of NCI-Black-Reiter male rat bladder carcinogenesis by dimethylarsinic acid an organic arsenic compound. AB - Dimethylarsinic acid (DMAA) is a major metabolite of inorganic arsenicals in mammals. In the present study, we investigated its promoting effects on urinary bladder carcinogenesis in NCI-Black-Reiter (NBR) rats, which lack alpha2u globulin synthesizing ability. Male 9-14-week-old NBR rats were treated sequentially with 0.05% N-butyl-N-(4-hydroxybutyl)-nitrosamine (BBN) for 4 weeks and then given 100 ppm DMAA in their drinking water (group 1) for 32 weeks. Induction of preneoplastic lesions (papillary or nodular hyperplasia) in this DMAA-treated group was significantly increased as compared to the carcinogen alone control group (P < 0.01). The development of carcinomas was also enhanced and a significant increase in the 5-bromo-2'-deoxyuridine (BrdU) labeling index of the urinary bladder epithelial cells was observed for the DMAA treatment group. These results indicate that DMAA has promoting effects on urinary bladder carcinogenesis even in NBR rats, so its effects are not dependent on the presence of alpha2u-globulin. PMID- 10381128 TI - Inhibitory effects of dehydrozingerone and related compounds on 12-O tetradecanoylphorbol-13-acetate induced Epstein-Barr virus early antigen activation. AB - Dehydrozingerone, 4-(4-hydroxy-3-methoxyphenyl)-3-buten-2-one, is half an analog of curcumin which is known to have anti-tumor activity. The anti-tumor promoting activity of dehydrozingerone was evaluated by determining the inhibitory effect on Epstein-Barr virus early antigen (EBV-EA) activation induced by 12-O tetradecanoylphorbol-13-acetate (TPA). The concentration needed for 50% inhibition of the tumor promotion (IC50) of dehydrozingerone was similar to that of curcumin. To elucidate the structure-activity relationship on the anti-tumor promoting activity, dehydrozingerone, curcumin, isoeugenol, which has no carbonyl group in the side chain, benzalacetone, which is the basic structure of dehydrozingerone, o-dehydrozingerone, which is the ortho-hydroxyl substituted compound of dehydrozingerone, and their related compounds were investigated using the in vitro short-term assay on TPA-induced EBV-EA activation. o Dehydrozingerone showed the most potent inhibitory effect in a series of tested dehydrozingerone derivatives and their related monosubstituted benzalacetones. This suggests that the occupation at both ortho positions of the hydroxyl group enhances the anti-tumor promoting activity. Isoeugenol inhibited the tumor promoting activity at a concentration of about one-third of the IC50 of dehydrozingerone. This indicates that the carbonyl group in the side chain has a negative impact on the anti-tumor promoting activity. The inhibitory effects of the carbon-carbon bond in the side chain were studied using benzylacetone with a single bond, benzalacetone with a double bond and 4-phenyl-3-butyn-2-one with a triple bond. 4-Phenyl-3-butyn-2-one inhibited the most potent activity followed by benzalacetone and benzylacetone. PMID- 10381129 TI - Inhibitory effects of uterine endometrial carcinogenesis in Donryu rats by tamoxifen. AB - The effects of tamoxifen (TAM) on uterine carcinogenesis were investigated in female Donryu rats. The effects were initiated by a single intrauterine treatment with N-ethyl-N'-nitro-N-nitrosoguanidine (ENNG) at a dose of 20 mg/kg body weight via the vagina at 10 weeks of age. TAM tubes (cholesterol tubes containing 50% TAM) were implanted into the backs of the rats for 13 months (full TAM group) or for the second-half of this period (half TAM group). In the control group treated with ENNG alone, various proliferative lesions were induced in the uterine endometrium and the incidence of endometrial adenocarcinomas was about 30%. In contrast, the uteri in both TAM-treated groups showed severe atrophy and the incidences of uterine proliferative lesions were limited to a few endometrial hyperplasias in the half TAM group. Most of the vaginas in both TAM-treated groups showed mucification, while cornification was common in the vaginal epithelium of controls. The ovaries demonstrated similar atrophy with cystic follicles and no corpora lutea in all groups. Other estrogen responsive endocrine organs, such as the pituitaries and adrenals, were small in the TAM-treated groups. Serum estrogen levels in the TAM-treated groups were lower than in the control group but progesterone levels did not differ. These results indicated that TAM acts as an anti-estrogen on the adult rat uterus, inhibiting the development of endometrial adenocarcinomas initiated by ENNG. PMID- 10381130 TI - Differentiation-inducing effect of retinoic acid, difluoromethylornithine, sodium butyrate and sodium suramin in human colon cancer cells. AB - We investigated the relative effectiveness of four differentiation-inducing chemicals to induce a more normal or benign phenotype in the human colon cancer cell lines Moser and HT29. The differentiation-inducing capability of all-trans retinoic acid (ATRA), difluoromethylornithine (DFMO), sodium butyrate (NaB) and sodium suramin (NaS) was evaluated in terms of the efficacy of these chemicals in inhibiting cellular proliferation, growth in soft agarose, invasion of matrigel and induction of morphological alteration. The relative ability of these chemicals to induce production of the differentiation-related molecules fibronectin and carcinoembryonic antigen was also determined. Overall, ATRA was found to be the most effective chemical in inducing differentiation as measured by these parameters. The Moser cells were more susceptible to differentiation induction by comparison with the HT29 cells. Both similarities and differences in the cellular responses to DFMO, NaB and NaS were also observed for the Moser and HT29 cells. The differences in cellular responses to these chemicals may be due to different phenotypic properties of these two cell lines and different mechanisms of action of these chemicals. PMID- 10381131 TI - Cloning genes responsive to a hepatocarcinogenic peroxisome proliferator chemical reveals novel targets of regulation. AB - To better understand the molecular basis of the hepatocyte proliferation and induction of hepatocellular adenomas by exposure to peroxisome proliferator chemicals (PPC), a systematic search for genes modulated by a PPC (WY-14643) in rat liver was carried out using the differential display technique. The fragments fell into two classes based on the time of initial and maximal induction by WY 14643. The class I genes (clones 5 and 30) were induced 3 h after a gavage exposure to WY-14643 with maximal expression at 24 h. The class II genes (clones 13 and 16) were induced after 24 h with maximal expression at 78 weeks. Expression of the class II genes was also increased after other treatments that cause cell proliferation. Clone 30 was identified as CYP4A2, previously shown to be regulated by PPC. Clone 13 was homologous to the mouse protein H gene, a component of the heterogeneous nuclear ribonucleoprotein particle important in mRNA splicing. Clone 16 was identified as cyclophilin-A, the receptor for the immunosuppressant drug cyclosporin A. The sequence of clone 5 was unique. These data demonstrate that WY-14643 increases the levels of a number of novel genes that are coordinately regulated with increases in chronic cell proliferation and fatty acid metabolism. PMID- 10381132 TI - Protective effects of diallyl sulfide on N-nitrosodimethylamine-induced immunosuppression in mice. AB - Diallyl sulfide (DAS), a flavor component of garlic that has been used as a food additive, exerts chemopreventive effects at several organ sites in rodents after administration of chemical carcinogens possibly by inhibiting carcinogen activation via cytochrome P450-mediated oxidative metabolism. In this study, we investigated the protective effect of DAS on the N-nitrosodimethylamine (NDMA) induced immunosuppression of humoral and cellular responses in BALB/c mice and the possible mechanisms involved in this protection. We observed that oral administration of DAS prior to NDMA treatment for 14 consecutive days blocked the NDMA-induced suppression of the antibody response to a T-cell-dependent antigen, sheep erythrocytes, and the lymphoproliferative response to the T-cell and the B cell mitogens in dose-dependent manners. Treatment of mice with DAS resulted in a significant decrease of cytochrome P450 2E1-dependent p-nitrophenol hydroxylase and NDMA demethylase activities. The results show that the protective effects of DAS against the NDMA-induced immunotoxicity may, at least in part, be due to its ability to block bioactivation of NDMA mainly by the inhibition of cytochrome P450 2E1. PMID- 10381133 TI - Effects of resveratrol on 12-O-tetradecanoylphorbol-13-acetate-induced oxidative events and gene expression in mouse skin. AB - Resveratrol (3,5,4'-trihydroxy-trans-stilbene) is a natural product shown to inhibit carcinogen-induced pre-neoplastic lesions in mouse mammary organ culture and 12-O-tetradecanoylphorbol-13-acetate (TPA)-promoted mouse skin tumors. Application of TPA to mouse skin induces oxidative stress, as evidenced by numerous biochemical responses, including significant generation of H2O2 and enhanced levels of myeloperoxidase and oxidized glutathione reductase activities and decreases in glutathione levels and superoxide dismutase activity. TPA treatment also elevates the expression of cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), c-myc, c-fos, c-jun, transforming growth factor-beta1 (TGF-beta1) and tumor necrosis factor-alpha (TNF-alpha). As currently reported, pre-treatment of mouse skin with resveratrol negated several of these TPA-induced effects in a dose-dependent manner. H2O2 and glutathione levels were restored to control levels, as were myeloperoxidase, oxidized glutathione reductase and superoxide dismutase activities. As judged by reverse transcriptase-polymerase chain reaction (RT-PCR), TPA-induced increases in the expression of c-fos and TGF beta1 were selectively inhibited. These data suggest that resveratrol inhibits tumorigenesis in mouse skin through interference with pathways of reactive oxidants and possibly by modulating the expression of c-fos and TGF-beta1. PMID- 10381134 TI - Antitumour promoting activity of indole-3-carbinol in mouse skin carcinogenesis. AB - There has been growing interest in recent years in the potential of brassica vegetables (cabbage, cauliflower, brussels sprouts, etc.) as vectors for the introduction of anticarcinogenic compounds in the diet. Indole-3-carbinol, a major indole metabolite present in the cruciferous vegetables, has been found to inhibit various rodent tumours when administered prior to or during carcinogen exposure. In this study, the antitumour promoting potential of indole-3-carbinol was studied in a two-stage mouse skin model of carcinogenesis. The animals were initiated with a single subcarcinogenic dose of DMBA. After one week, 250 microg of indole-3-carbinol was applied topically to each animal prior to promotion with 5 microg TPA twice per week. Tumour development was significantly inhibited in indole-3-carbinol-supplemented animals in terms of cumulative numbers of tumours and average tumours per mouse. About 44% of male and 29% of female mice remained tumour-free in this group at the end of the experiment. A significant delay in the tumour induction time was also observed in indole-3-carbinol-supplemented animals. This evidence suggests that indole-3-carbinol, in the manner and dose given, inhibits the development of tumours in the two-stage mouse skin model of carcinogenesis. PMID- 10381135 TI - In vitro changes of the nuclear AgNORs pattern induced by RNA inhibitors and 5 fluorouracil in human breast cancer cells, MCF-7 and HBC-4. AB - The morphological changes of argyrophilic nucleolar organizer regions (AgNORs) were studied in two human breast cancer cell lines, MCF-7 and HBC-4. Treatment with an RNA polymerase inhibitor (actinomycin D) reduced the size of AgNORs and increased the number of AgNORs. Messenger RNA polymerase inhibitor (alpha amanitin) also increased the number of AgNORs. However, translational blocking agents closely related to ribosomal RNA (cycloheximide and anisomycin) caused a decrease in the number of AgNORs, which seemed to fuse to an aggregate around the nucleolus and formed a single large spherical AgNOR in the final stage. These changes were observed typically when cells were treated with 5-fluorouracil or 5 fluorouridine. These morphological changes in the AgNORs pattern, AgNORs aggregation, might reflect certain damage in ribosomal RNA. PMID- 10381136 TI - Phenobarbital-induced cytoplasmic accumulation of beta1-integrin in rat liver enzyme altered foci; an immunohistological study. AB - In this immunohistological study we investigated integrin expression in EAF in female rats treated with diethylnitrosamine (DEN) as initiator and phenobarbital (PB) as promotor (DEN-PB treatment) for up to 32 weeks. Using a beta1-integrin antibody, there was an increased cytoplasmic staining and a decreased sinusoidal staining in EAF, as compared to non-EAF areas. The majority of small EAF and all larger EAF exhibited this altered distribution of beta1-integrin. The increased cytoplasmic staining was not found in EAF after a 10 week treatment-free period. In periportal areas in partial hepatectomized control rats a similar increase in cytoplasmic staining was seen. EAF in DEN-initiated and DEN-promoted rats (DEN DEN treatment) were also studied. This protocol induced rapidly growing EAF. Most lesions did not show the increased cytoplasmic staining. However, after partial hepatectomy of DEN-DEN-treated rats, a cytoplasmic staining was seen in EAF. It is concluded that PB induced a reversible cytoplasmic beta1-integrin expression in many EAF and in all larger EAF. It is suggested that the alteration constitutes part of hepatocyte resistance to toxicological stress and apoptosis in EAF. PMID- 10381137 TI - In vitro effects of gonadotropin-releasing hormone (GnRH) analogue on cancer cell sensitivity to cis-platinum. AB - Six endometrial cancer cell lines (Ishikawa, EIIL, HEC1A, 6, 50 and 59), one breast cancer cell line (MCF-7) and two ovarian cancer cell lines (OVHS-1, HRA) were treated for 24 or 168 h with a gonadotropin-releasing hormone (GnRH) analogue, Buserelin acetate, and the cellular growth profile was studied. All these cell lines except for the HRA line had positive GnRH receptor mRNA expression detected by reverse transcriptase polymerase chain reaction. GnRHa suppressed cell growth after 168 h of exposure, but not after 24 h. Suppression of cell growth by the exposure to cis-platinum (CDDP, 10 nM for 24 h) was significantly increased in the presence of GnRHa for 168 h. The mechanism of this growth inhibition was tested by examining both RNA components of human telomerase (hTR) expression and telomerase activity. The results showed that GnRHa inhibits telomerase activity without altering the RNA component of telomerase expression. The present data suggest that GnRH analogue may modulate endometrial, breast and ovarian cancer cell growth through modifying the telomerase activity. Since GnRHa increased the cytotoxic effects of CDDP and GnRHa is a compound of high patient compliance, the value of GnRHa as a tumor sensitizer to CDDP should be further tested in clinical trials. PMID- 10381138 TI - Serum leptin is associated with serum uric acid concentrations in humans. AB - This cross-sectional study aimed to evaluate the relationship between leptin and the cluster of abnormalities often referred to as the metabolic syndrome. The serum leptin concentration, body mass index (BMI), percent body fat, total fat mass (FM), waist and hip circumference, waist to hip ratio (WHR), prevalence of hypertension, and triglyceride (TG), lipoprotein, and uric acid concentration were determined in 86 type 2 diabetic (n = 59) and healthy (n = 27) subjects. Multiple regression analyses showed that the estimates of total body obesity (BMI, percent body fat, and total FM), sex, and serum uric acid concentration are independently associated with the serum leptin concentration. The finding of a positive correlation between serum leptin and uric acid levels suggests that leptin could be a pathogenic factor responsible for hyperuricemia in obesity. PMID- 10381139 TI - Functional and molecular expression of intestinal oligopeptide transporter (Pept 1) after a brief fast. AB - The intestinal oligopeptide transporter, cloned as Pept-1, has major roles in the assimilation of dietary proteins and absorption of peptidomimetic medications. The initial aim of the present experiment was to investigate whether the functional expression of this transporter is affected by dietary intake. Functional expression was determined as the rate of uptake of glycylglutamine (Gly-Gln) by brush-border membrane vesicles (BBMVs) prepared from the jejunum of fed and fasted rats. Surprisingly, the rate of dipeptide uptake was greatly increased after 1 day of fasting. The subsequent aim of the experiment became the investigation of the mechanism of this alteration in transport, which showed that 1 day of fasting increased (1) the maximal Gly-Gln uptake (Vmax) by twofold without changing the Km of Gly-Gln uptake by BBMVs, (2) the amount of intestinal oligopeptide transporter (Pept-1) protein by threefold in the brush-border membrane, and (3) the abundance of Pept-1 mRNA by threefold in the intestinal mucosa. We conclude that 1 day of fasting increases dipeptide transport in rat intestine by increasing the population of Pept-1 in the brush-border membrane. The mechanism appears to be an increase in Pept-1 gene expression. PMID- 10381140 TI - Pitfalls in the use of 2-octynoic acid as an in vivo model of medium-chain acyl coenzyme A dehydrogenase deficiency: ketone turnover and metabolite studies in the rat. AB - 2-Octynoic acid was administered by intraperitoneal injection to fasted Sprague Dawley rats in an attempt to simulate medium-chain acyl-coenzyme A dehydrogenase (MCAD) deficiency. The resultant urine organic acid profile showed a mild dicarboxylic aciduria but lacked the glycine conjugates characteristic of MCAD deficiency. Further studies with infused 13C(4)-3-hydroxybutyrate and 13C(2) acetoacetate demonstrated reduced ketone production in treated rats compared with control animals. Although plasma ketone body concentrations were low in treated rats, plasma free fatty acids were also low, thereby providing diminished substrate for ketone production. This is the reverse of the finding in children with MCAD deficiency, who have low levels of plasma ketones despite elevated free fatty acids. These animal studies were therefore not helpful in improving our understanding of ketone body kinetics in children with MCAD deficiency. PMID- 10381141 TI - Inhibition by retinoids of benzo(A)pyrene metabolism catalyzed by 3 methylcholanthrene-induced rat cytochrome P-450 1A1. AB - Benzo(a)pyrene, a well-known procarcinogen, is believed to be activated by microsomal cytochrome P-450 1A1 (CYP 1A1). We recently reported that rat CYP 1A1 induced by 3-methylcholanthrene (3-MC) catalyzed the conversion of retinal to retinoic acid. In this study, we investigated retinoid inhibition of the metabolism of benzo(a)pyrene and 7-ethoxyresorufin, another specific substrate of CYP 1A1, using liver microsomes prepared from control and 3-MC-pretreated rats as the enzyme source. In 3-MC-treated rats, retinal and retinol, but not retinoic acid, inhibited benzo(a)pyrene metabolism. The 50% inhibitory concentration (IC50) of retinal was about 11.5 micromol/L and the inhibition was competitive, with a Ki value of 5.8 micromol/L. Retinol is a more potent inhibitor than retinal. The IC50 was about 5 micromol/L and the inhibition was mixed, with a Ki value of 19.2 micromol/L and a Ki' value of 4.2 micromol/L. Almost the same results were obtained for the reaction of 7-ethoxyresorufin deethylation. In contrast, the metabolic activity of both benzo(a)pyrene and 7-ethoxyresorufin was much lower in untreated versus 3-MC-treated rats, and only weak inhibition by the retinoids was observed. The results suggest that retinoids inhibit the activation of benzo(a)pyrene and that the substrate specificity of cytochrome P-450 isozymes associated with retinoid metabolism should be taken into account when studying the anticarcinogenic action of retinoids. PMID- 10381142 TI - Role of chloride and inhibitory action of inorganic nitrate on gonadotropin stimulated steroidogenesis in mouse Leydig tumor cells. AB - The involvement of adenylate cyclase-cyclic adenosine monophosphate (AC-cAMP) in gonadotropin-stimulated testicular steroidogenesis is well known. Little is known about the role of guanylate cyclase-cyclic guanosine monophosphate (GC-cGMP) or early chloride conductance stimulated by gonadotropins in steroidogenesis. Human chorionic gonadotropin (hCG) 1 IU/L caused significant androgen secretion without a discernible effect on cAMP production. Despite negligible intracellular cAMP, the protein kinase A inhibitor H89 blocked basal and hCG-stimulated steroidogenesis. The GC inhibitors methylene blue (MB) and LY83583 decreased androgen secretion, but hCG did not stimulate cGMP production and there was not a steroidogenic response to exogenous cGMP. A chloride-channel inhibitor, diphenylamine-2-carboxylate (DPC), at concentrations up to 0.6 mmol/L stimulated basal steroid secretion and hCG 10 IU/L stimulated cAMP production, but higher concentrations had an inhibitory effect. Substitution of chloride by gluconate enhanced basal steroid secretion, but nitrate completely abolished the effect of 1 IU/L hCG on androgen secretion, which could be partially overcome by increasing the gonadotropin concentration. In conclusion, chloride, perhaps by activating AC cAMP, mediates the steroidogenic action of gonadotropins in mouse Leydig tumor cells (MLTC-1). Inorganic nitrate probably inhibited steroidogenesis via conversion to nitric oxide (NO) without involving the GC-cGMP pathway. Nevertheless, the results obtained with GC inhibitors suggest a role for the GC cGMP pathway in Leydig cell steroidogenesis. PMID- 10381143 TI - Glucose tolerance and insulin resistance in the JCR:LA-corpulent rat: effect of miglitol (Bay m1099). AB - A standardized meal tolerance test (MTT) using 5 g rat chow provides a sensitive index of insulin and glucose metabolism in the insulin-resistant, hyperinsulinemic, hypertriglyceridemic, and atherosclerosis-prone JCR:LA corpulent (cp) strain of rats. The MTT revealed differences in insulin/glucose metabolism that were not evident in either an intravenous (IVGTT) or intraperitoneal (IPGTT) glucose tolerance test. The glycemic response of control rats to a 5-g carbohydrate test meal containing miglitol (Bay m1099) was sharply reduced, with a 50% effective dose (ED50) of 36.4 +/- 7.5 mg/100 g food. At a dose of 60 mg/100 g food, the plasma glucose curve was flat and indistinguishable from that found in the nonfed state. The plasma insulin response was similarly reduced, with an ED50 of 42.8 +/- 14.8 mg/100 g food. Obese male rats were treated with miglitol at 60 mg/100 g food from 6 to 12 weeks of age. Treated rats had a significantly reduced food consumption and lower body weight at 12 weeks of age (P < .05). The treatment resulted in no significant changes in serum lipid concentrations. When subjected to the MTT using control (non-miglitol-containing) food, treated rats demonstrated markedly improved insulin sensitivity, with a greatly reduced insulin response, which may reflect an improved hepatic glucose metabolism. The results confirm that miglitol is highly effective in this obese insulin-resistant animal model. It reduced postprandial glycemic and insulin responses, and on long-term treatment, it improved glucose and insulin metabolism. These beneficial metabolic changes suggest that miglitol may have vascular protective effects in insulin-resistant states. PMID- 10381144 TI - Substrate interactions in the short- and long-term regulation of renal glucose oxidation. AB - The present study evaluated the substrate competition between fatty acids (FA) and glucose in the kidney in vivo in relation to the operation of the "glucose FA" and "reverse glucose-FA" cycles. In fed rats, neither inhibition of adipocyte lipolysis by 5-methylpyrazole-3-carboxylic acid (MPCA) nor inhibition of mitochondrial long-chain FA oxidation by 2-tetradecylglycidate (TDG) influenced the renal ratio of free/acylated carnitine or the percentage of total renal pyruvate dehydrogenase complex (PDHC) in the active (dephosphorylated) form (PDHa). The additional provision of glucose, a precursor for the synthesis of malonyl-coenzyme A (coA), did not influence renal PDHa activity or the renal ratio of free to acylated carnitine, implying that FA oxidation is maximally suppressed in the fed state. A reverse glucose-FA cycle may therefore be important in suppressing renal FA oxidation in the fed state. After 48 hours of starvation, MPCA and TDG decreased short- and long-chain acylcarnitine concentrations (40% to 50%, P < .01) and elevated the renal ratio of free/acylated carnitine (2.5-fold, P < .001, and 3.3-fold, P < .001, respectively), indicating that FA oxidation is increased after starvation. Despite suppression of renal FA oxidation, renal PDHa activity in 48-hour starved rats was only partially restored by treatment with MPCA or TDG. The additional administration of glucose did not remedy this. The failure to reverse completely the effects of prolonged starvation in suppressing PDHC activity by acute inhibition of FA oxidation suggests additional regulatory mechanisms that dampen the PDHC response to acute changes in substrate supply. Estimations of PDH kinase (PDK) activity in renal mitochondria showed a significant 1.7-fold stable increase (P < .01) after 48 hours of starvation. Analysis of PDK pyruvate sensitivity in renal mitochondria incubated with respiratory substrate (5 mmol/L 2-oxoglutarate/0.5 mmol/L L-malate) showed that the pyruvate concentration required for 50% activation was substantially decreased by starvation. Enzyme linked immunosorbent assay (ELISA) analysis over a range of PDHC activities demonstrated that increased PDK activity was concomitant with a significant (at least P < .01) 1.8-fold increase in the protein expression of the ubiquitously expressed PDK isoform, PDK2. We hypothesize that changes in protein expression and activity of individual PDK isoforms may dictate the renal response to incoming FA lesterification v oxidation) through modulation of the relationship between glycolytic flux and PDHC activity, and thus the provision of precursor for malonyl-coA production. PMID- 10381145 TI - Properties of native and in vitro glycosylated forms of the glucagon-like peptide 1 receptor antagonist exendin(9-39). AB - The intestinal hormone glucagon-like peptide-1-(7-36)-amide (GLP-1) has recently been implicated as a possible therapeutic agent for the management of type 2 non insulin-dependent diabetes mellitus (NIDDM). However, a major difficulty with the delivery of peptide-based agents is their short plasma half-life, mainly due to rapid serum clearance and proteolytic degradation. Using a peptide analog of GLP 1, the GLP-1 receptor antagonist exendin(9-39), we investigated whether the conjugation of a carbohydrate structure to exendin(9-39) would generate a peptide with intact biological activity and improved survival in circulation. The C terminal portion of exendin(9-39) was reengineered to generate an efficient site for enzymatic O-glycosylation. The modified exendin(9-39) peptide (exe-M) was glycosylated by recombinant UDP-GalNAc:polypeptide N acetylgalactosaminyltransferase 1 (GalNAc-T1) alone or in conjunction with a recombinant GalNAc alpha2,6-sialyltransferase (Sialyl-T), resulting in exe-M peptides containing either the monosaccharide GalNAc or the disaccharide NeuAc alpha2,6GalNAc. The nonglycosylated and glycosylated forms of exe-M competed with nearly equal potency (> 90% of control) with the binding of [125I]GLP-1 to human GLP-1 receptors expressed on stably transfected COS-7 cells. In addition, each peptide was equally effective for inhibiting GLP-1-induced cyclic adenosine monophosphate (cAMP) production in vitro. Studies with rats demonstrated that the modified and glycosylated forms of exendin(9-39) could antagonize exogenously administered GLP-1 in vivo. Interestingly, glycosylated exendin(9-39) homologs were more than twice as effective as the nonglycosylated peptide for inhibiting GLP-1-stimulated insulin production in vivo, suggesting a longer functional half life in the circulation for glycosylated peptides. Results from in vivo studies with 3H-labeled peptides suggest that the glycosylated peptides may be less susceptible to modification in the circulation. PMID- 10381146 TI - Synergistic interaction of magnesium and vanadate on glucose metabolism in diabetic rats. AB - The effect of vanadate (V) alone, magnesium (Mg) alone, and the combination of Mg plus V (MgV) on insulin-mediated glucose disposal and glucose tolerance was investigated in normal and streptozotocin-induced diabetic rats. MgV, magnesium sulfate (MgSO4) and sodium metavanadate (NaV) were added to the drinking water of normal or diabetic rats (approximately 300 g) for 3 weeks. After 3 weeks of V treatment (both MgV and NaV), diabetic rats demonstrated a normal meal tolerance test without any increase in the plasma insulin response. Rats also received a euglycemic insulin clamp (12 mU/kg x min for 120 minutes) with 3-3H-glucose infusion to quantify total body glucose disposal, glycolysis (3H2O production), and glycogen synthesis (total body glucose disposal minus glycolysis). Total glucose disposal was decreased in diabetic versus control rats (29 +/- 2 v 35 +/- 2 mg/kg x min, P < .01) and returned to levels greater than the nondiabetic control values after MgV (41 +/- 2, P < .01). Supersensitivity to insulin was not observed in diabetic rats treated with NaV (34 +/- 1). Glycogen synthesis was increased by both MgV and NaV treatment (23 +/- 21, P < .01 and 18 +/- 1, P < .05 v 14 +/- 2 mg/kg x min) in diabetic rats. A small increase in glycolysis was observed in MgSO4 and MgV rats (18 +/- 1 and 18 +/- 1 v 16 +/- 1, P < .05). NaV alone had no effect on glycolysis. Thus, Mg has a synergistic effect with V to increase muscle glycogen synthesis in diabetic rats. In normal rats, neither MgSO4 nor NaV had any effect on glucose utilization. However, MgV increased glucose disposal to rates that were significantly higher than the rate in untreated control rats (P < .05). Based on these results, MgV is superior to either V alone or Mg alone in improving insulin sensitivity and glycogen synthesis in diabetic rats. PMID- 10381147 TI - Isotopic enrichment of amino acids in urine following oral infusions of L-[1 (13)C]phenylalanine and L-[1-(13)C]lysine in humans: confounding effect of D [13C]amino acids. AB - Urine sampling of the free amino acid pool serves to reflect plasma enrichment and is used as a noninvasive means to determine isotope enrichment in studies of amino acid metabolism. We determined the effect of D-[13C]phenylalanine and D [13C]lysine content of tracers on urinary amino acid enrichment following oral infusion of L-[13C]phenylalanine in 18 preterm infants and L-[1-(13)C]lysine in seven healthy adult females. Urinary [13C]phenylalanine enrichment was higher (P < .0001) for L-[13C]phenylalanine containing 0.4% D-[13C]phenylalanine (28.6 +/- 7.1) versus L-[1-(13)C]phenylalanine that contained undetectable D [13C]phenylalanine (10.2 +/- 1.5). D-[13C]phenylalanine, measured by chiral column gas chromatography-mass spectrometry (GC-MS), accounted for 10% to 30% (20.5% +/- 7%) of total phenylalanine in the urine of infants who received 0.4% D [13C]phenylalanine, and was absent from the urine of infants receiving tracer with undetectable [13C]phenylalanine. Urinary L-[13C]phenylalanine enrichment did not differ between tracer groups (9.8 +/- 1.5 and 9.8 +/- 2.5). In adult females, the use of L-[1-(13)C]lysine (1.6% D-lysine) resulted in a higher (P < .02) urine total L,D-[13C]lysine enrichment compared with plasma enrichment (40.8 +/- 4.1 v 11.1 +/- 0.7). This study demonstrates the significant presence of D-[13C]amino acids in urine that originate as contaminants from commercially manufactured tracers, as a result of renal tubular discrimination of D-amino acids. A tracer containing detectable amounts of D-[13C]isomer cannot be recommended for any study in which urine will be used to reflect enrichment in the arterial plasma pool. PMID- 10381149 TI - Leptin serum levels are not correlated with disease activity in patients with rheumatoid arthritis. AB - Leptin, the ob gene product, has been proposed as a mediator of inflammatory cytokine-dependent decreased food intake and cachexia in rodents. In humans, leptin serum levels increase after administration of tumor necrosis factor-alpha (TNF-alpha) or interleukin-2 or during septicemia. However, the effect of human chronic inflammatory disease on serum leptin is unknown. We therefore determined the serum leptin level (radioimmunoassay), body mass index (BMI), percent body fat ([%BF] bioelectrical impedance analysis), and disease activity (Disease Activity Score [DAS]) in 58 patients with rheumatoid arthritis (RA) and 16 controls. The BMI, %BF, serum leptin, and ratio of leptin to %BF (leptin/%BF) did not differ significantly in 25 patients with moderate RA activity (DAS, 3.6 +/- 0.5), 33 patients with low RA activity (DAS, 1.8 +/- 0.5), and controls. A positive correlation for serum leptin and %BF was detected in all groups. Our data indicate that in RA, a human chronic cytokine-mediated inflammatory disease, the serum leptin level is directly related to %BF but not to disease activity. PMID- 10381148 TI - Effect of reduced inspired oxygen on fetal growth and maternal glucose metabolism in rat pregnancy. AB - The effect of prolonged exposure to a reduced fraction of inspired oxygen ([FiO2] 0.17 for 3 days) on maternal glucose kinetics, placental glucose transporters GLUT1 and GLUT3, and fetal growth was examined in rat pregnancy. Arterial and venous catheters were placed 3 days before the study. [3-(3)H]glucose tracer and deuterium labeling of water were used to measure the rates of glucose turnover and gluconeogenesis (GNG), respectively. Glucose uptake by maternal tissues was measured using [14C]2-deoxyglucose. Exposure to a reduced FiO2 resulted in a significant decrease (mean +/- SE) in fetal weight (room air, 4.02 +/- 0.04 g; 0.17 FiO2, 3.27 +/- 0.6 g, P < .02). There was a significant increase in the maternal-fetal glucose gradient (maternal-fetal glucose ratio: room air, 1.48 +/- 0.11; 0.17 FiO2, 2.26 +/- 0.24, P < .05), but there was no change in the maternal or fetal blood lactate concentration. No significant change in maternal blood pH was observed; however, a significant decrease in the blood partial pressure of O2 (PO2) occurred (room air, 97 +/- 0.5 torr; 0.17 FiO2, 81 +/- 1.8) on day 3. There was no change in the rate of turnover of glucose or GNG in the maternal compartment, nor was there any effect on glucose uptake by the maternal tissues. Placental GLUT1 and GLUT3 mRNA were not different in the control or experimental animals. We conclude that a mild reduction in the FiO2 for 3 days in rat pregnancy results in a significant fetal growth restriction that is not related to any observed alteration in maternal glucose metabolism. The lower glucose concentration in the fetal blood may be the consequence of an increase in fetal glucose metabolism, thereby resulting in an increased maternal-fetal gradient of glucose. PMID- 10381150 TI - Black-white differences in postprandial triglyceride response and postheparin lipoprotein lipase and hepatic triglyceride lipase among young men. AB - Black-white differences in serum triglycerides and high-density lipoprotein (HDL) cholesterol concentrations are known. However, the metabolic basis for these differences is not clear. This study determined the magnitude of postprandial triglyceride concentrations, lipoprotein lipase and hepatic triglyceride lipase activities in postheparin plasma, and serum lipid and lipoprotein cholesterol concentrations in healthy young adult black men (n = 22) and white men (n = 28). Postprandial triglyceride concentrations were measured at 2, 3, 4, 5, 6, and 8 hours after a standardized test meal. Serum lipid and lipoprotein cholesterol concentrations were similar between the races in this study sample. However, incremental (above basal) increases in triglycerides were significantly greater in white men versus black men at 2 hours (P = .01) and tended to be greater at 3 hours (P = .12) and 4 hours (P = .06) after the fat load. In a multivariate analysis that included age, race, apolipoprotein E (apoE) genotype, fasting triglycerides, obesity measures, alcohol intake, and cigarette use, fasting triglycerides (P = .04) and, to a lesser extent, race (P = .07) were associated independently with the 2-hour incremental increase in triglycerides. The incremental triglyceride response correlated inversely with HDL cholesterol in both whites (r = -.38, P = .04) and blacks (r = -.59, P = .004). Lipoprotein lipase activity was higher (P = .049) and hepatic triglyceride lipase activity lower (P = .0001) in black men compared with white men; racial differences persisted after adjusting for the covariates. While lipoprotein lipase activity tended to associate inversely with the postprandial triglyceride concentration in both races, hepatic triglyceride lipase activity tended to correlate positively in whites and inversely in blacks. These results suggest that compared with whites, blacks may have an efficient lipid-clearing mechanism that could explain the black-white differences in lipoproteins found in the population at large. PMID- 10381151 TI - Effects of different insulin infusion rates on heart rate variability in lean and obese subjects. AB - The low-frequency to high-frequency ratio (LF/HF ratio) is an index of cardiac sympathovagal balance. We hypothesized that insulin might also stimulate the LF/HF ratio. Thus, 15 lean and 15 obese subjects were studied. Each subject underwent sequential hyperinsulinemic clamps (insulin infusion rate 0.50, 1, and 2 mU/kg x min) while the heart rate was recorded by the Holter technique continuously. Indirect calorimetry allowed determination of the respiratory quotient (Rq) and substrate oxidation. The leg blood flow (LBF), leg vascular resistance (LVR), and plasma norepinephrine concentration were also measured. In seven lean subjects, hyperinsulinemic clamps were repeated along with propranolol infusion (0.1 mg x kg(-1) as an intravenous bolus dose followed by continuous intravenous infusion of 0.5 mg x kg(-1) x min(-1) throughout the study). Lean subjects had better insulin action than obese subjects. Insulin infusion was associated with an increase of the deltaLF/HF ratio in both lean (P < .001 for time-dependent changes) and obese (P < .02 for time-dependent changes) subjects; however, the extent of insulin-mediated stimulation of the LF/HF ratio was greater in lean versus obese subjects. Insulin infusion did not significantly affect HF values in both groups. Independently of gender, body fat, changes in the plasma norepinephrine concentration, LBF, and LVR, the deltaLF/HF ratio at the end of the fastest insulin infusion (0.8 +/- 0.2 v 0.3 +/- 0.2, P < .04) was still greater in lean versus obese subjects. The deltaLF/HF ratio was also more stimulated during insulin versus insulin + propranolol infusion in lean subjects. In conclusion, insulin stimulates the LF/HF ratio in both lean and obese subjects and thus produces a shift in the cardiac autonomic nervous system activity toward sympathetic predominance. PMID- 10381152 TI - Growth hormone corrects acidosis-induced renal nitrogen wasting and renal phosphate depletion and attenuates renal magnesium wasting in humans. AB - We have shown previously that chronic hyperchloremic metabolic acidosis (CMA) induces severe negative nitrogen balance and renal phosphate depletion and decreases serum insulin-like growth factor-1 (IGF-1) in association with growth hormone (GH) insensitivity in humans. The present study investigated whether acidosis-induced renal nitrogen wasting and renal phosphate depletion are mediated by GH insensitivity/low IGF-1 and thereby responsive to GH treatment. The effects of GH on acidosis-induced changes in divalent cation metabolism and acidosis-induced hypothyroidism were also investigated. CMA (delta[HCO3], -10.5 mmol/L) was induced in six healthy male subjects ingesting 4.2 mmol NH4Cl/kg body weight [BW]/d for 7 days. Recombinant human GH (0.1 U/kg BW/12 h subcutaneously) was administered for 7 days while acid feeding was continued. GH increased serum IGF-1 from 22.1 +/- 1.4 to 87 +/- 8.4 nmol/L (control level, 36.4 +/- 2.2). GH decreased urinary nitrogen excretion, resulting in a cumulative nitrogen retention of 2,404 mmol, thereby correcting the acidosis-induced cumulative increase in nitrogen excretion (2,506 mmol) despite continued acid feeding. GH attenuated the acidosis-induced hyperphosphaturia (cumulative phosphate retention, 91 mmol) and corrected the hypophosphatemia. GH did not affect acidosis-induced ionized hypercalcemia, but further exacerbated acidosis-induced hypercalciuria (cumulative loss, 27.3 mmol). GH significantly further increased serum 1,25-dihydroxyvitamin D (1,25(OH)2D) and further decreased intact PTH (from 10 +/- 1 to 6 +/- 1 pg/mL). Acidosis also induced hypomagnesemia and hypermagnesuria (cumulative loss, 9.4 mmol, ie, renal magnesium wasting), a novel finding, which was significantly attenuated by GH (cumulative retention, 5.0 mmol). In conclusion, GH corrected acidosis-induced renal nitrogen wasting, which may be caused, at least in part, by decreased IGF-1 levels. GH further increased serum 1,25(OH)2D and the systemic calcium load, which account for the suppression of parathyroid hormone (PTH) despite renal PO4 retention and correction of hypophosphatemia. GH attenuated acidosis-induced renal magnesium wasting. PMID- 10381153 TI - Elevated atrial natriuretic peptides and early renal failure in type 2 diabetic Goto-Kakizaki rats. AB - The present investigation was designed to determine if atrial natriuretic peptides (ANPs) are increased in a spontaneous model of non-obese type 2 diabetes, the Goto-Kakizaki (GK) rat. Four peptide hormones originating from the ANP prohormone were increased twofold (P < .05) to sixfold (P < .01) in the circulation of GK rats compared with nondiabetic Wistar rats from which the GK colony was originally derived. Thus, ANP, long-acting natriuretic peptide (LANP), vessel dilator, and kaliuretic peptide were (mean +/- SE) 497 +/- 78, 1,285 +/- 105, 457 +/- 45, and 385 +/- 87 pg/mL in GK rats, versus 78 +/- 23, 542 +/- 77, 137 +/- 26, and 134 +/- 33 pg/mL, respectively, in Wistar rats. In evaluating the cause of the increased ANPs, the blood volume of GK rats (16.2 +/- 0.4 mL) was significantly (P < .01) increased compared with Wistar rats (9.5 +/- 0.3 mL). The ventricles of GK rats were not dilated when examined by transthoracic echocardiography, but the venous system was markedly distended. GK rats had a 48% to 79% decrease in renal function (ie, increased serum creatinine and blood urea nitrogen [BUN]) compared with Wistar rats. These results indicate that circulating ANPs are increased in the GK spontaneously diabetic rat secondary to (1) increased blood volume, which leads to increased synthesis and release of ANPs, and (2) renal failure, which results in a delayed metabolic processing of these peptides. The early combined increases of the four atrial peptides collectively may contribute to the hyperfiltration that occurs in early diabetes mellitus. PMID- 10381154 TI - Leukocyte glycolysis and lactate output in animal sepsis and ex vivo human blood. AB - Lactate is released in large quantity from sites of sepsis and inflammation. We asked whether the increased lactate production found in sepsis can be explained by the augmented glycolysis of inflammatory cells. The glycolytic metabolism of rat peritoneal leukocytes was measured following cecal ligation and perforation (CLP) or sham laparotomy. CLP augmented glucose uptake, the pentose phosphate pathway, and glucose oxidation. Lactate output increased from 1.03 +/- 0.05 to 1.20 +/- 0.05 fmol x cell(-1) x min(-1) (P < .001). Total lactate output of peritoneal lavage fluid increased from 7.94 +/- 2.59 to 28.12 +/- 5.60 nmol L x min(-1) (P < .005). The effect of lipopolysaccharide (LPS) on the lactate output of whole blood from 31 critically ill patients was measured. Leukocyte lactate production was calculated by multiple linear regression analysis. Following exposure to LPS, human leukocyte lactate output increased from 0.20 +/- 0.09 to 1.22 +/- 0.14 fmol x cell(-1) x min(-1) (P < .001). This rate of production is so high that it suggests that the lactate output of different tissue beds in sepsis may be affected by their different cell populations and state of activation. This study supports the hypothesis that lactate may be more a product of inflammation than a marker of tissue hypoxia in sepsis. PMID- 10381156 TI - Short-term glucocorticoid administration decreases both hypothalamic and pituitary galanin synthesis in the adult male rat. AB - Galanin (GAL) is a peptide that has been implicated in the regulation of the growth axis. It is generally accepted that GAL can increase serum growth hormone (GH) levels, although the underlying mechanism for this increase is unknown. It is well known that long-term glucocorticoid treatment alters in vivo GH secretion, since there is a decrease in serum GH in response to stimuli. It has previously been shown in our laboratory that administration of GAL can overcome the effects of glucocorticoid administration on GH secretion. The aim of the present study was to determine the effects of long-term glucocorticoid administration on the regulation of hypothalamic and pituitary GAL mRNA levels. Adult male rats were treated for 72 hours with the synthetic glucocorticoid dexamethasone ([DEX] 40 microg/kg/d intraperitoneal injections). RNase protection assays were performed on both the hypothalamus and pituitary for the presence of GAL mRNA. As expected, DEX significantly decreased somatic growth, as evidenced by a decrease (50%) in the weight gain of glucocorticoid-treated versus control animals. It was also demonstrated that in both the hypothalamus and pituitary, glucocorticoid treatment reduced the level of GAL mRNA (to 11% and 6.5%, respectively) compared with the control condition. We conclude that the decrease in GAL mRNA may lead to a decrease in GAL secretion, which in turn may be involved in the glucocorticoid-induced inhibition of GH secretion. PMID- 10381155 TI - 15-Deoxy-delta(12,14) prostaglandin J2: a putative endogenous promoter of adipogenesis suppresses the ob gene. AB - Leptin is considered a key factor in the regulation of appetite and energy expenditure, but little is known about the control of its synthesis and release. Thiazolidinediones (TZDs) have recently been shown to downregulate leptin expression, and it has been speculated that downregulation of the ob gene occurs through activation of the transcription factor, peroxisome proliferator-activated receptor gamma (PPARgamma). However, there are no studies using an endogenous PPARgamma ligand. We examined the effect of 15-deoxy-delta(12,14) prostaglandin J2 (15d-PGJ2), a putative natural ligand of PPARgamma, on ob gene expression in fully differentiated 3T3-L1 adipocytes and compared its effect with that of two other PPARgamma activators, the TZD troglitazone (Trog) and indomethacin (Indo). 15d-PGJ2, Trog, and Indo all inhibited leptin expression at concentrations at which they activate PPARgamma. The inhibition of leptin expression of PPARgamma activators was surprising, since PPARgamma is known to induce adipogenesis during which the ob gene is expressed. To address the possibility that PPARgamma plays different roles before and after the induction of adipogenesis, we examined the effects of the three PPARgamma ligands on the expression of leptin and the glucose transporter protein GLUT4, both of which are expressed during differentiation of 3T3-L1 preadipocytes to adipocytes. In the absence of PPARgamma ligands, leptin and GLUT4 synthesis increased from day 3 to day 9 or 10 during differentiation. However, in the presence of any of the three PPARgamma ligands, GLUT4 expression was unaffected, while ob gene expression was inhibited. We hypothesize that PPARgamma may be essential for induction of adipocyte differentiation but then needs to be inactivated to allow expression of the ob gene. PMID- 10381157 TI - Chronic ethanol consumption leads to destabilization of rat liver beta galactoside alpha2,6-sialyltransferase mRNA. AB - Chronic ethanol consumption in rats is accompanied by decreased levels of Gal beta1,4GlcNAc alpha2,6-sialyltransferase (2,6-ST) activity in the liver. Our previous studies have shown that there is a concomitant decrease in the levels of 2,6-ST mRNA. In this study, the alteration in the regulation of 2,6-ST expression by chronic ethanol consumption was assessed by Northern hybridization, nuclear run-on experiments, and 2,6-ST mRNA stability studies. 2,6-ST downregulation was found at 4 weeks of feeding an ethanol diet (36% of calories from ethanol) and remained up to 8 weeks. The decrease in 2,6-ST mRNA levels was found to be dose dependent, with lower dose of ethanol (12% and 24% of total dietary calories from ethanol) being ineffective and the effects being manifested only when 36% of the dietary calories were from ethanol. The effects of chronic ethanol feeding could be completely reversed within 1 week after ethanol consumption was stopped, when 2,6-ST mRNA levels were restored to normal. The downregulation was not sensitive to actinomycin D, indicating that the regulation was not affected at the transcriptional level but at the posttranscriptional level. This was confirmed by nuclear run-on experiments showing that the rate of 2,6-ST mRNA transcription was unaffected by ethanol. Finally, mRNA stability experiments showed that the half life of 2,6-ST mRNA was reduced 50% in ethanol-fed rat livers compared with control rat livers. Taken together, the results show that 2,6-ST mRNA is regulated at the posttranscriptional level and chronic ethanol intake downregulates 2,6-ST expression by destabilizing its mRNA. PMID- 10381158 TI - Effects of cigarette smoking and its cessation on body weight and plasma leptin levels. AB - Smokers weigh less than age-matched nonsmokers, and most smokers gain weight after smoking cessation due to an increase in food intake and a decrease in energy expenditure. Leptin is an endocrine signal thought to regulate body fat stores through hypothalamic control of energy intake and expenditure. To determine whether the "weight-reducing" effects of smoking may be mediated by leptin, we measured plasma leptin concentrations in 22 middle-aged and older male smokers (body mass index [BMI], 28 +/- 1 kg/m2, mean +/- SEM) and 22 nonsmokers matched to the smokers for age (64 +/- 1 years) and BMI (28 +/- 1 kg/m2). The body weight and leptin concentration were remeasured at 3 and 6 months in 13 of the smokers who successfully stopped smoking. The leptin concentration correlated positively with the BMI in both smokers (r = .74, P < .001) and nonsmokers (r = .76, P < .001). However, the intercept of the regression line was higher for smokers versus nonsmokers (P < .05), with no difference in the slope. Thus, male smokers have a higher leptin level for a given BMI than nonsmokers. Following 6 months of smoking cessation, body weight increased by 7% (6.0 +/- 0.1 kg, n = 13, P < .01). Despite this weight gain, the mean leptin concentration did not increase with smoking cessation. On average, leptin levels were 25% lower than would be expected for the amount of weight gained after smoking cessation. These findings suggest that cigarette smoking directly elevates circulating plasma leptin concentrations, and this increase may be one mechanism for the lower body weight of smokers compared with nonsmokers. PMID- 10381160 TI - Brain mechanisms in fatal cardiac arrhythmia. AB - Intravenous injection of histamine to rabbits was used as a prototype in an investigation of the mechanism of sudden death due to anaphylaxis and other causes. The "dive" reflex, bradycardia due to activation of the ophthalmic branch of the fifth cranial nerve, was induced in thirty-seven of the animals while they inhaled a very small amount of cigarette smoke. Associated with the resulting bradycardia were lowered blood pH and increased serum content of lactic acid and potassium and increased peripheral arterial constriction with elevation of diastolic blood pressure. Intravenous injection of 1 ml of histamine in the presence of the dive reflex induced potentially fatal ventricular arrhythmia, but no cardiac disturbance when administered while the dive reflex was inactive, thereby strongly suggesting that sudden death in anaphylaxis may involve an overzealous response to a normally protective neural reflex. PMID- 10381159 TI - Combined effect of vegetable protein (soy) and soluble fiber added to a standard cholesterol-lowering diet. AB - Dietary treatment of hyperlipidemia focuses on reducing saturated fat and dietary cholesterol. Other aspects of diet are not emphasized at present, despite growing evidence that a number of plant components decrease serum cholesterol. We therefore determined whether a combination of two plant components, vegetable protein and soluble fiber, further reduce serum lipids when incorporated into the currently advocated low-saturated-fat diet. Thirty-one hyperlipidemic men and women ate two 1-month low-fat (<7% of total energy from saturated fat), low cholesterol (<80 mg cholesterol/d) metabolic diets in a randomized crossover study. The major differences between test and control diets were an increased amount of vegetable protein (93% v 23% of total protein), of which 33 g/d was soy, and a doubling of soluble fiber. Fasting blood samples were obtained at the start and end of each phase. On the last 3 days of each phase, fecal collections were obtained. Compared with the low-fat control diet, the test diet decreased total cholesterol (6.2% +/- 1.2%, P < .001), low-density lipoprotein (LDL) cholesterol (6.7% +/- 1.7%, P < .001), apolipoprotein B (8.2% +/- 1.2%, P < .001), and the ratios of LDL to high-density lipoprotein (HDL) cholesterol (6.3% +/- 2.0%, P = .004) and apolipoprotein B to A-I (5.4% +/- 1.5%, P = .001). A combination of vegetable protein and soluble fiber significantly improved the lipid-lowering effect of a low-saturated-fat diet. The results support expanding the current dietary advice to include increased vegetable protein and soluble fiber intake so that the gap in effectiveness between a good diet and drug therapy is reduced. PMID- 10381161 TI - Idioventricular low frequency oscillation in QT interval responds univocally to RR confusing kinds of mental stress. AB - Seventeen male subjects, aged nineteen to twenty, went through a protocol including, while supine, relaxation at rest (10 min) and mental stress (MS) by a Kraepelin (arithmetic) test (5 min), as part of a larger study. With a polygraphic analog recording set-up we also collected a 1 ms - digital facsimile of a lead II-like thoracic ECG with maximum T-wave (Codas, Dataq Instr). Twenty three stress responses were assigned to three classes according to known cardiotacho-, plethysmo-, and pneumo-graphic marks of "concentrated attention mainly," "emotion," or still "high emotion." During each setting the most stationary 3 min RR epoch in cardiotachogram was selected for joint RR & QT beat by-beat variability study. RR and QT intervals were detected using a published algorithm. Conventional RR and QT Fourier autospectra were computed, while using RR*QT mean square coherence spectrum we detached the RR-independent, idioventricular (IV) fraction of QT low frequency (LF: 0.04-0.15 Hz) power of variability (IV QT-LF). IV QT-LF responded consistently to varieties of mental stress that confuse RR-LF or let QT-LF unchanged, best witnessing the cortically issued ventricular adrenergic strain. Indeed, while emotion propels the same way all spectral variables above, concentrated attention increased (Wilcoxon) significantly IV QT-LF only (0.54-0.80 ms2) and decreased RR-LF (715-463 ms2). Findings hold promise of a non-invasive, high resolution Holter based monitoring of sympathetic status of myocardium, robust vis-a-vis of confusion caused by the autonomic interplay at sino-atrial node. PMID- 10381162 TI - Nonaware classical conditioning to pictorial facial stimuli in a between-groups paradigm. AB - The present experiment investigated the effects of aware and nonaware modes of extinction in classical conditioning to facial emotional stimuli. The subjects participated in three different experimental phases. In the first (habituation) phase they were presented with a 500 ms angry face. In the second (acquisition) phase, for half of the subjects the 500 ms face was paired with an aversive noise (experimental group) while for the other half of the subjects the face and the noise presentations were separated by 6-10 s intervals (sensitization control group). In the third (extinction) phase, these two groups were further divided into two subgroups. One subgroup of both the experimental and control group had the face stimulus presented for 30 ms, and immediately masked with a neutral picture. The other two subgroups had the face presented for 500 ms with no mask. The results showed that conditioning only occurred in the experimental subgroups which was indicated by a significant difference between skin conductance responses during habituation and corresponding responses during extinction. Secondly, comparing the experimental and control groups during the extinction phase, a significant conditioning effect was observed for both the aware and nonaware masked modes of extinction for the experimental group. The results suggest that conditioned autonomic responses may be elicited in a nonaware mode. PMID- 10381163 TI - Psychophysiological correlates of organizational change and threat of unemployment among police inspectors. AB - The study examined psychosocial work-conditions and physiological reactions among thirty-six police inspectors (median age 45 years, 81% males) who participated in a reorganization. At this time, subjects were threatened by unemployment and had to re-apply for their positions in a new police district. Data were collected during the reorganization and at three years follow-up, by means of questionnaires (Stress Profile) and blood samples. The blood samples were used to determine serum levels of gamma-glutamyltransferase (GGT), glucose, lipids, prolactin, testosterone and cortisol. The results show a positive association between worry about employment and symptoms of burnout during the reorganization. Mean scores for the Stress Profile sub-scales worry about employment (p<.01) and work-load (p<.05) decreased between measurements, but an impairment in relationships with management was noticed (p<.05). No significant changes were observed in terms of self-rated health complaints. Significant decreases in total cholesterol (p<.0001), LDL-cholesterol (p<.0001), LDL/HDL-ratio (p<.01), prolactin (p<.0001), as well as increases in testosterone (p<.01) and cortisol (p<.001) were observed for the whole sample. Glucose decreased with marginal significance (<.07). Controlling for age and gender, multivariate regression analyses showed that subjects who reported deteriorations in satisfaction with work manifested the most modest decreases in prolactin (p<.05). Also, the decrease in glucose was larger for subjects who experienced impairments in satisfaction with work (p<.05), information (p<.05), task-oriented leadership, (p<.05), and respect and dignity (p<.05). Subjects who perceived deteriorations in the ethical and moral standards of the organization increased their cortisol level to a lower degree than their counterparts (p<.05). Favorable changes in employment status and psychosocial work environment seem to be related to improved physiological functioning. PMID- 10381164 TI - A study of the effects of cranial electrical stimulation on attention and concentration. AB - There have been several anecdotal accounts that cranial electrical stimulation (CES) enhances attention and the ability to learn new tasks in a normal population, but only one published investigation confirms that CES improves attention using the Alpha Stim CES (Madden and Kirsch, 1987). The purpose of this study was to corroborate the findings of Madden and Kirsch, using more precise measures of attention, such as a Continuous Performance Test (CPT). A pretest and posttest CPT was given to two groups using the LISS CES device. The control group consisted of twenty-one subjects who received the placebo treatment. The experimental group of thirty-one subjects received twenty minutes of CES. Four measures of the CPT show significant gains in attention: Number of Hits, p =.010 Hit RT ISI Change, p =.016, Risk Taking, p =.055; and Attentiveness, p =.054. Based on subjects who demonstrated improvement by one standard deviation on two different measures of the CPT, thirty-one percent of the experimental group improved versus four percent of the control group. The use of CES as a method of increasing attention is a promising area that requires further investigation. PMID- 10381165 TI - Colonic sensitivity in irritable bowel syndrome and normal subjects according to their hemispheric preference and cognitive style. AB - According to our earlier results, non-painful, weak afferent visceral signals may exert a steady influence on brain processes, including cognitive functions. In the present series colonic impulses of irritable bowel syndrome (IBS) subjects served as a model of chronic impact from the gut. Hemispheric preference, as well as cognitive style of information processing served as indicators of covert changes in brain functions. In twenty-one IBS patients and in ten control subjects of both sexes, the thresholds of minimal colonic distension sensitivity has been measured following the determination of hemispheric preference and of advantage in verbal or spatial information processing of the subjects. In IBS patients distension thresholds proved to be higher in verbals than in spatials, whereas in healthy controls the relationship of colonic thresholds and verbal versus spatial advantage was reversed. Among the normal controls with left hemisphere preference a significantly higher distension threshold has been observed than in those with right hemisphere preference, whereas in the IBS group such threshold-differences were not observable. PMID- 10381166 TI - Serologic and genetic characterization of HIV type 1 subtypes on Reunion Island. AB - The aim of this study was to determine the HIV subtypes present on Reunion Island, a French island located in the Indian Ocean, where the first case of AIDS was diagnosed in 1987. Paired sera and blood samples were collected between September 1996 and September 1997 from 53 HIV-1-positive patients. Subtyping was performed by serotyping with a previously described subtype-specific enzyme immunoassay (SSEIA) and by genotyping with the heteroduplex mobility assay (HMA). When samples gave uninterpretable results with either of the methods, or discordant results, the V3 env region was sequenced and genetic subtypes were determined by phylogenetic analysis. Genetic subtyping showed that 48 of 53 patients were infected with HIV-1 subtype B (90.5%). This high prevalence of subtype B on Reunion Island is probably due to the regular exchanges with metropolitan France. The other five patients were infected with subtype A (9.5%); they had been directly linked to African populations. Of the 48 subtype B samples, 44 (91.7%) were correctly subtyped by SSEIA and 43 (89.6%) by HMA. However, the SSEIA did not allow the subtyping of A strains in three of five patients. Thus, the SSEIA could be an alternative routine technique for screening subtype B versus nonsubtype B HIV-1 strains. PMID- 10381167 TI - Intracellular activation of 2',3'-dideoxyinosine and drug interactions in vitro. AB - Didanosine (2',3'-dideoxyinosine; ddI) requires intracellular metabolism to its active triphosphate, 2',3'-dideoxyadenosine 5'-triphosphate (ddATP), to inhibit the replication of human immunodeficiency virus (HIV). We have investigated the metabolism of ddI to ddATP in the presence and absence of a range of compounds. In addition, we determined the levels of the endogenous competitor of ddATP, 2' deoxyadenosine 5'-triphosphate (dATP), and calculated ddATP/dATP ratios. None of the nucleoside analogs studied had any effect on ddI phosphorylation at 1 and 10 microM concentrations. At 100 microM concentrations, ddC reduced total ddA phosphates (82% of control total ddA phosphates; p < 0.001). ZDV significantly decreased the levels of dATP, whereas ddC significantly increased dATP pools (e.g., at 100 microM ZDV, 82% of control dATP levels; p < 0.001). Hence, the ddATP/dATP ratio was increased in the presence of ZDV, but was decreased in the presence of ddC. Neither d4T nor 3TC affected the ddATP/dATP ratio. Deoxyinosine (dI) significantly reduced ddA phosphate production at 100 microM concentrations, with ddATP reduced to undetectable levels (p < 0.001). Hydroxyurea (HU) did not affect the activation of ddI, but significantly reduced dATP pools at 100 microM concentrations (67% of control dATP levels; p < 0.001), enhancing the ddATP/dATP ratio. ddA phosphate production was significantly reduced by pentoxyfylline (PXF) at 10 and 100 microM concentrations. dATP levels were unaffected, but the ddATP/dATP ratio was reduced. Finally, 8-aminoguanosine (8-AMG) had no effect on either ddI activation or dATP pools. These studies demonstrate the importance of determining both the active TP and the competing endogenous TP, as changes to the resulting ratio could alter the efficacy of the nucleoside analog in question. PMID- 10381168 TI - Downregulation of Bcl-2, but not of Bax or Bcl-x, is associated with T lymphocyte apoptosis in HIV infection and restored by antiretroviral therapy or by interleukin 2. AB - The role of Bcl-2, Bax, and Bcl-x in the apoptosis of T lymphocytes in HIV infected individuals was investigated. A strong correlation between Bcl-2 downregulation and spontaneous apoptosis has been reported by various groups in short-term cultures of CD8+ but not of CD4+ T lymphocytes. We describe a similar correlation in CD4+ T cells and provide an explanation why Bcl-2 downregulation in these cells has not been detected so far. In apoptotic cells not only Bcl-2, but also the CD4 surface receptors, are downregulated, preventing the detection of these cells in flow cytometric analysis. In contrast to Bcl-2, no correlation is detectable between Bax or Bcl-x expression and apoptosis. T lymphocytes of HIV infected, but not of control, individuals display ex vivo a heterogeneous Bcl-2 expression pattern with a low and a high Bcl-2-expressing lymphocyte fraction. The proportion of low Bcl-2-expressing T cells correlates with a higher viral load in these individuals. Antiretroviral therapy significantly reduces the proportion of low Bcl-2-expressing lymphocytes, which is associated with a decrease in apoptosis. Bcl-2 downregulation and spontaneous apoptosis of T lymphocytes from HIV-infected individuals can be partially prevented by the exogeneous addition of IL-2, but not of IL-12, IL-4, or antibodies that prevent the CD95/CD95 ligand pathway of apoptosis. PMID- 10381170 TI - HTLV type I Tax activation of the CXCR4 promoter by association with nuclear respiratory factor 1. AB - Human T lymphotropic virus type I trans-activator Tax protein regulates expression of several cellular genes that are involved in cellular activation, proliferation, and transformation. Tax mediates its regulatory activity through interaction with cellular transcription factors such as members of the cAMP responsive element-binding factors/ATF family or the NF-kappaB/Rel family. In this study we have demonstrated that Tax trans-activates the promoter for CXCR4, a coreceptor for T cell-tropic HIV-1 through its association with nuclear respiratory factor 1 (NRF1). The promoter region for CXCR4 contains an NRF1 binding site, which is crucial for basal and Tax-induced activity. Glutathione S transferase (GST) pull-down experiments showed association of GST-Tax fusion protein with NRF1 in vitro. Expression of Tax, in addition to stimulation with phorbol myristate acetate and ionomycin, increased formation of the NRF1 complex in a gel-mobility shift assay, indicating that Tax association with NRF1 in vivo facilitates its DNA binding. HTLV-I Tax activation of CXCR4 may contribute to the rapid progression of HIV disease observed in certain coinfected individuals. PMID- 10381169 TI - Independent evolution of HIV type 1 in different brain regions. AB - HIV-1-associated brain pathology exhibits regional variability and we therefore studied the genetic differences in the V1-V5 domains of the HIV env gene in up to four regions of brain (frontal lobe, basal ganglia, medial temporal lobe, and nonmedial temporal lobe) from three patients. We found that in each separate brain region HIV-1 forms different quasispecies and that there is little gene flow among these regions. In further support of brain region-specific evolution of HIV-1, we analyzed amino acid signatures in these clones. In addition to known amino acid signatures associated with macrophage tropism and the lack of syncytium formation, we found 15 majority amino acid signature patterns from the V1-V5 env sequences associated with the neuroanatomical regions analyzed from the three individuals. Furthermore, on average, intrabrain genetic distances for the HIV-1 env were estimated to be much smaller than genetic distances between brain regions. Specific strains of HIV-1 may be neurotropic or neuroinvasive (replication preference in brain tissue) and may contribute to pathology, cognitive loss, and neuropsychiatric disease. PMID- 10381171 TI - Effect of insulin-like growth factor I on HIV type 1 long terminal repeat-driven chloramphenicol acetyltransferase expression. AB - In this study, we have investigated the ability of insulin-like growth factor I (IGF-I) to inhibit HIV long terminal repeat (LTR)-driven gene expression. Using COS 7 cells cotransfected with tat and an HIV LTR linked to a chloramphenicol acetyltransferase (CAT) reporter, we observed that physiological levels of IGF-I (10(-9) M) significantly inhibited CAT expression in a concentration- and time dependent manner. IGF-I did not inhibit CAT expression in COS 7 cells transfected with pSVCAT, and did not affect CAT expression in the absence of cotransfection with tat. Transfection of HIV-1 proviral DNA into COS 7 cells +/- IGF-I resulted in a significant decrease (p < 0.05) in infectious virion production. Both IGF-I and Ro24-7429 inhibited LTR-driven CAT expression, while TNF-alpha-enhanced CAT expression was not affected by IGF-I. On the other hand, a plasmid encoding parathyroid hormone-related peptide exhibited dramatic additivity of inhibition of CAT expression in COS 7 cells. Finally, we show that in Jurkat or U937 cells cotransfected with HIVLTRCAT/tat, IGF-I significantly inhibited CAT expression. Further, interleukin 4 showed in U937 cells inhibition of CAT expression that was not additive to IGF-I induced inhibition. Our data demonstrate that IGF-I can specifically inhibit HIVLTRCAT expression. This inhibition may occur at the level of the tat/TAR interaction. Finally, this IGF-I effect is seen in target cell lines and similar paths of inhibition may be involved in the various cell types employed. PMID- 10381172 TI - Immune activation and virologic response to immunization in recent HIV type 1 seroconverters. AB - Antigenic stimulation from invasive bacterial infections, and the vaccines designed to prevent them, may promote T cell activation and enhancement of HIV-1 replication. Changes in viral load have been correlated with antigen-specific responses. We prospectively determined the impact of immunization with 23-valent pneumococcal vaccine (PVAX) and Haemophilus influenzae type b (Hib)-modified diphtheria toxoid CRM197 (DT) vaccine on HIV-1 replication in recent HIV-1 seroconverters (n = 14; median, 5.5 months from infection; median CD4+ T cells, 535 microl), and correlated results with vaccine-related immune activation. Specific antibody responses, markers of CD4+ T cell activation (transferrin and interleukin 2 receptors), and viral burden were measured at weeks -2 (pre), 0, 1, 2, 6, and 12 after immunization. By week 2, levels of IgG had increased significantly over baseline in both HIV-1-infected patients and HIV-1 seronegative control subjects (n = 9) for each antigen (geometric mean fold rise: PVAX, 10.1 versus 5.3; Hib, 16.0 versus 11.7; and DT, 26.2 versus 24.5, respectively). Despite these vigorous responses to both polysaccharide and protein antigens, HIV-1-infected patients showed limited evidence of CD4+ T cell activation at 1 week, no consistent rise in HIV-1 burden at any point, and no decline in CD4+ T cell number over time. We conclude that recent HIV-1 seroconverters show vigorous humoral responses to vaccine antigens and limited early evidence of T cell activation, but no substantial or sustained increase in viral replication or decline in CD4+ T cell number. Thus, respiratory bacterial vaccines appear immunogenic and safe early in HIV-1 infection. PMID- 10381173 TI - Antibody-dependent cellular cytotoxicity in HIV type 1-infected patients receiving VaxSyn, a recombinant gp160 envelope vaccine. AB - Antibody-dependent cellular cytotoxicity (ADCC) activity was measured in 60 human immunodeficiency virus (HIV-1)-infected patients receiving a recombinant gp160 (rgp160) envelope protein of HIV-1(NL4-3) in alum and 64 receiving placebo over a 5-year study period. There was no difference in the percentage of ADCC responders when comparing rgp160-immunized patients (mean, 78.4%) with those receiving placebo alone (mean, 81.5%) at any time point examined. Patients were further divided into progression groups regardless of their vaccine status. ADCC activity was somewhat higher in rapid than in slow-progressing groups, although the number that had detectable ADCC activity was equivalent in each group. ADCC activity of sera from rapid- and slow-progressing groups against primary or laboratory isolate envelopes was similar. This study showed that transcription with rgp160 did not appear to enhance HIV-specific ADCC activity. ADCC activity did not appear to correlate with protection against AIDS in this cohort of HIV-1-infected people. PMID- 10381174 TI - Inhibition of the anti-V3 loop response to a recombinant gp120SF2 vaccine by preexisting monoclonal antibody. AB - Our previous studies have shown that maternally transferred MAb 83.1 (Repligen), which recognizes the V3 loop of HIV-1SF2, inhibited the anti-V3 IgG response of offspring BALB/c mice immunized with rgp120SF2 (Chiron). To determine the mechanism of this epitope-specific inhibition, MAb 83.1 was directly administered intraperitoneally to 19-day-old BALB/c mice, followed by immunization with rgp120SF2 in Freund's complete adjuvant at 21 days of age. The total serum anti rgp120SF2 IgG response was not inhibited by preexisting MAb 83.1, but the anti-V3 IgG response was significantly inhibited (p < 0.03) 12 weeks after immunization. These results confirm epitope-specific inhibition of the immunodominant V3 loop and are consistent with epitope masking by preexisting antibody. Idiotypic dysregulation is unlikely since MAb 83.1 exposure was not required during the period of repertoire development in neonatal mice. Treatment with MAb prior to immunization may redirect the epitope specificity of an HIV vaccine response. PMID- 10381175 TI - Cytotoxicity and superoxide anion generation by some naturally occurring quinones. AB - Four naturally occurring quinones, mansonone-D (MD), mansonone-H (MH), thespone (TP) and thespesone (TPE), extracted from the heartwood of Thespesia populnea have been tested for their cytotoxic action by aerobic incubation with human breast adenocarcinoma (MCF-7) cells. Toxicity of the quinones follows the order MD > TP > MH approximately TPE. EPR spectrometric and Clark electrode oximetric studies indicate that redox cycling of these quinones produce superoxide anion radical (O2*-) and H2O2 on aerobic incubation with NADH:cytochrome c reductase. Generation of superoxide radical during enzymatic reduction of quinones, was confirmed by EPR spin trapping experiment using 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) as a spin trap. Cyclic voltammetric studies show reversible redox couples for MD and TP whereas TPE and MH show irreversible redox couple. The electrochemical results indicate that MH and TPE are more difficult to reduce than TP and MD. PMID- 10381176 TI - Induced mono-(ADP)-ribosylation of rat liver cytosolic proteins by lipid peroxidant agents. AB - We have studied the effect of free radical generating agents on the mono-(ADP) ribosylation of rat liver cytosolic proteins. Our results show that this post translational modification, whose physiological significance is still unclear, is activated by lipid peroxidant agents via activation of cytoplasmatic mono-(ADP) ribosyltransferases. The implication of free radicals in this process is demonstrated by the fact that mono-(ADP)-ribosylation can be prevented by melatonin, N-tert-butyl-alpha-phenylnitrone and dithiothreitol. On the basis of our results, we discuss the modification of proteins caused by free radicals as a possible mechanism by which they damage the cell. PMID- 10381177 TI - Free radical activity following contraction-induced injury to the extensor digitorum longus muscles of rats. AB - The purpose of the study was to investigate the role of free radicals in the injury induced by a protocol of repeated pliometric (lengthening) contractions to the extensor digitorum longus (EDL) muscle in situ in rats. Previous data have indicated that prior treatment with the antioxidant polyethylene glycol superoxide dismutase reduced the damage that was apparent at 3 days following this type of exercise. Three hours and 3 days following the protocol, the magnitude of the semiquinone-derived free radical signal observed by electron spin resonance spectroscopy (ESR) was not different for exercised and non exercised skeletal muscles. A reduction in the protein thiol content of muscle was evident at 3 h, and was still apparent at 3 days. Three hours after the protocol, the total muscle glutathione content and the percentage in the oxidized form were unchanged, but by 3 days the percentage of muscle glutathione present in the oxidized form was elevated. The susceptibility of muscle to lipid peroxidation in vitro was reduced 3 days after the pliometric contractions. These data indicate that oxidation of protein thiols and glutathione may be involved in the secondary damage following pliometric contractions, but provide no evidence that the species involved were derived from mitochondrial semiquinone radicals. PMID- 10381178 TI - Role of different Ca2+ sources in the superoxide production of human neutrophil granulocytes. AB - The role of different Ca2+ sources in the activation of the NADPH oxidase was investigated in human neutrophil granulocytes. Selective depletion of the stimulus-responsive intracellular Ca2+ -pool and the consequent opening of the store-dependent Ca2+ channel of the plasma membrane was achieved with thapsigargin, an inhibitor of microsomal Ca2+ -ATPase. Low concentration (10-100 nM) of thapsigargin did not induce any O2*- -production, indicating that elevation of [Ca2+]ic to similar level and probably via similar route as following stimulation of chemotactic receptors, by itself is not sufficient to activate the NADPH oxidase. In significantly higher concentration (1-10 microM) thapsigargin did induce O2*- -generation but this effect was not the result of elevation of [Ca2+]ic. In the absence of external Ca2+ a gradual decrease of the responsive Ca2+ pool was accompanied by a gradual decrease of the rate and duration of the respiratory response stimulated by formyl-methionyl-leucyl phenylalanin. Maximal extent of receptor-initiated O2*- -production could only be obtained when the intracellular [Ca2+] was higher than the resting level. Under this condition Ca2+ originating from intracellular or external source was equally effective in supporting the biological response. PMID- 10381179 TI - Scavenging effect of benzophenones on the oxidative stress of skeletal muscle cells. AB - Benzophenone is an ultraviolet (UV)-absorbing agent that has been used in industry and medicine for more than 30 years. Consumers of cosmetics and sunscreens containing UV-absorbers are exposed to benzophenones on a daily basis, owing to the widespread use of these compounds. However, the efficacy of these compounds as scavengers of oxidative stress is still not well established. In the present study, we investigate the antioxidative capacity of six sunscreen benzophenone compounds. A primary myoblast culture was mixed in vitro with 100 microM menadione. The cytotoxic effect by menadione-induced oxidative stress was monitored by the lucigenin- or luminol-amplified chemiluminescence, methylthiotetrazole (MTT) assay, and the antioxidative effects of various benzophenone compounds were evaluated. The results showed that the addition of menadione can induce oxidative stress on myoblasts by superoxide and hydrogen peroxide production, which can be eradicated by superoxide dismutase (SOD) and catalase, respectively, in a dose-dependent mode. The catalase has a protective effect on the cytotoxicity induced by menadione as measured by the MTT assay, while the SOD does not. The selected benzophenones also have a significant scavenging effect on the menadione-induced cell death on the myoblasts. The ortho dihydroxyl structure and other hydroxy groups in the same ring have a stronger scavenging effect on the superoxide anion on myoblasts; thus, a stable penoxy radical may be formed. The mechanism of this effect remains to be clarified. PMID- 10381180 TI - Liver AP-1 activation due to carbon tetrachloride is potentiated by 1,2 dibromoethane but is inhibited by alpha-tocopherol or gadolinium chloride. AB - Experimental acute intoxication by prooxidant haloalkanes produces marked stimulation of hepatic lipid peroxidation and cytolysis, which is followed by tissue regeneration. Our aim was to clarify the role of oxidative imbalance in the activation of the redox-sensitive transcription factor, activator protein-1 (AP-1), which is involved in tissue repair. Rats were poisoned with a very low concentration of carbon tetrachloride, given alone or in combination with another hepatotoxin, 1,2-dibromoethane, to provide varying extents of oxidative damage. The level of AP-1-DNA binding was analyzed by electrophoretic mobility shift assay on liver extracts, obtained from rats killed 6 h after poisoning. Stimulation of lipid peroxidation and AP-1 upregulation were already established when the hepatic damage due to carbon tetrachloride +/-1,2-dibromoethane was beginning to appear. Rat supplementation with the antioxidant vitamin E completely inhibited AP-1 upregulation, thus supporting a causative role of membrane lipid oxidation in the observed modulation of the transcription factor. Moreover, activation of Kupffer cells appears to be a crucial step in the increased AP-1 binding to DNA, the latter being largely prevented by gadolinium chloride, a macrophage-specific inhibitor. PMID- 10381181 TI - Effect of lecithinized superoxide dismutase (PC-SOD) on experimental pulmonary metastasis in mice. AB - The inhibitory effect of lecithinized superoxide dismutase (PC-SOD) on pulmonary metastasis in mice was investigated. In an experimental pulmonary metastasis model employing Meth A-T cells, significant and dose-dependent inhibition was observed after i.v. pre-administration of PC-SOD. Unmodified SOD (U-SOD) was also effective, but a 10-times higher dose was necessary to be significant. The pulmonary accumulation of Meth A-T cells labeled with 5-[125I]iodo-2' deoxyuridine was not reduced by either PC-SOD or U-SOD, and neither of the compounds decreased pulmonary MPO activity. However, PC-SOD increased pulmonary SOD activity for longer, compared with U-SOD. In vitro addition of PC-SOD dose dependently suppressed the growth of Meth A-T cells, while U-SOD had little effect. The combination of PC-SOD and S-nitroso-N-acetyl-D,L-penicillamine (SNAP), a nitric oxide (NO)-generating agent, had an additive effect. It was also found that PC-SOD prevented a decrease of pulmonary NOx level following tumor cell inoculation. It was concluded that PC-SOD possessed antimetastatic activity, and its potency was superior to that of U-SOD. These results suggest that PC-SOD may prevent the excessive formation of oxygen radicals and peroxynitrite (ONOO-) which cause cell damage and facilitate tumor metastasis. PMID- 10381182 TI - Increases in cholesterol 7-hydroperoxides in lipids of human skin by sunlight exposure. AB - Free and ester forms of cholesterol 7alpha- and 7beta-hydroperoxides (Ch 7-OOHs) in skin lipids of humans were separated and determined by high performance liquid chromatography with a chemiluminescence detector. We first demonstrated the presence of Ch 7-OOHs in lipids of human skin. The levels of Ch 7-OOHs found in skin lipids of healthy Japanese volunteers (n = 5) ranged from 2.78 to 25.2 pmol/cm2 skin, indicating large inter-individual differences. However, the intra individual differences of Ch 7-OOHs levels in skin lipids between right and left arms were less than 25% (-16.4% to 24.0%). Inter-day differences of Ch 7-OOHs in 5 subjects at 1 week interval were also small (-36.7% to 47.7%). Additionally, we investigated effects of sunlight exposure on the levels of Ch 7-OOHs in skin lipids of healthy Japanese volunteers (n = 24). The levels of Ch 7-OOHs in skin lipids significantly increased from 10.0+/-6.7 to 38.9+/-38.0 pmol/cm2 skin by sunlight exposure (10-40 mJ/cm2/min) for 3 h. Therefore, natural sunlight exposure causes lipid peroxidation in skin lipids of humans. These results suggest that the level of Ch 7-OOHs is a good marker for lipid peroxidation in human skin. PMID- 10381183 TI - Formation of the paramagnetic complex [Cr(OH)5O2]5- in the reaction between chromium(VI) oxide, and hydrogen peroxide or superoxide anion radicals studied by EPR spectroscopy. AB - Hydrogen peroxide or superoxide anion radicals form a paramagnetic complex in the reaction with chromium(VI) oxide in an alkaline water solution at room temperature. The complex [Cr(OH)5O2]5- with the g-value equal to 1.9734 is believed to contain hydroxyl groups derived from the alkaline solution and dioxygen derived from hydrogen peroxide or superoxide anion radicals. PMID- 10381184 TI - Effects of pyrrolidine dithiocarbamate on endothelial cells: protection against oxidative stress. AB - The dithiocarbamates are well known for their antioxidant properties and effects on cellular transcriptional events. For example, pyrrolidine dithiocarbamate (PDTC) is widely used as an inhibitor of nuclear factor kappa B (NFkappaB) and this, or related compounds may have therapeutic potential in inhibiting atherosclerosis. However, the precise molecular mechanisms through which PDTC could elicit antioxidant or cell signaling effects in a cellular setting remain unclear. Furthermore, the mechanisms for the effects of PDTC on NFkappaB are likely to involve inhibition of binding of the transcription factor to DNA rather than an effect on the activation process as first proposed. In relation to pharmacological applications of such compounds, little is known of their interaction with endothelial cells, the anticipated site of action for inhibition of vascular related diseases. Until recently, PDTC was generally classified as an antioxidant but evidence for pro-oxidant effects have been reported. In this study, we have addressed this issue in bovine aortic endothelial cells and identified two mechanisms through which PDTC can exert antioxidant effects. At low concentrations (0-25 microM), PDTC induces a concentration dependent increase in cellular GSH levels through the increased activity of gamma-glutamylcysteine synthetase. At higher concentrations, GSH oxidation and apoptotic cell death occur. Using 2,3 dimethoxy-1,4-napthoquinone (DMNQ) as an intracellular generator of superoxide radicals, we find PDTC (10 microM) protects against the cytotoxicity of this agent through a GSH-independent mechanism. PMID- 10381185 TI - Effect of structural modifications on photosensitizing activities of hypocrellin dyes: EPR and spectrophotometric studies. AB - Mono-substituted hypocrellin B (MHB) and di-substituted hypocrellin B (DHB) by mercaptoacetic acid are new photosensitizers synthesized to improve the red absorption and water solubility of the parent hypocrellin B (HB). The photochemistries (Type I and/or Type II) of MHB and DHB have been studied in homogeneous solutions using electron paramagnetic resonance (EPR) and spectrophotometric methods. In anaerobic homogeneous DMSO solution, DHB*- (or MHB*-) was predominantly photoproduced via self-electron transfer between the excited- and ground-state species. The presence of an electron donor significantly promotes the formation of the reduced form of DHB (or MHB). As compared with hypocrellin B, the efficiencies of DHB*- and MHB*- generation was enhanced obviously. When oxygen-saturated solutions of DHB (or MHB) were illuminated with 532 nm light, singlet oxygen (1O2), superoxide anion radical (O2*-), hydroxyl radical (*OH) and hydrogen peroxide (H2O2) were formed. DHB and MHB generate 1O2 with quantum yields of 0.18 and 0.22, respectively, which are much lower than that of HB (0.76) in chloroform. The superoxide anion radical was significantly enhanced by the presence of electron donors. The rate of O2*- production was also dependent on the concentration of DHB or MHB. Moreover, O2*- upon disproportionation can generate H2O2 and ultimately the highly reactive *OH via the Fenton reaction and other pathway with the involvement of DHB*- (or MHB* ). As in the case of DHB*- (or MHB*-), the efficiencies of O2*- and *OH generation by DHB and MHB were also enhanced obviously, consistent with the fact that DHB*- (or MHB*-) acts as the precursor of O2* and thus *OH. These findings suggest that the photodynamic actions of DHB and MHB may proceed via enhanced Type I mechanism and reduced Type II mechanism as compared with that of HB. PMID- 10381186 TI - L-carnitine attenuates doxorubicin-induced lipid peroxidation in rats. AB - Doxorubicin (DOX) was administered intraperitoneally to rats in six equal, 2.5 mg/kg doses over a 2-week period with or without L-carnitine. Injury was monitored by echocardiography, release of myosin light chain-1 (MLC-1), and by measurement of aldehydic lipid peroxidation products. General observation revealed that DOX alone caused more ascites than DOX plus L-carnitine. Animals sacrificed 2 h after the sixth dose had significantly higher aldehyde concentrations than 2 h after a single dose of DOX. Aldehydes in plasma and heart remained elevated for 3 weeks after the final dose of DOX, whereas L-carnitine prevented or attenuated the DOX-induced increases in lipid peroxidation. The increase in MLC-1 2 h after the sixth dose of DOX was greater than after a single dose, suggesting cumulative damage. Echocardiography did not detect either early injury or the protective effects of L-carnitine. These data indicate that lipid peroxidation following DOX occurs early, and parallels the cumulative characteristics of DOX-induced cardiotoxicity. The protective effects of L carnitine may be due to improved cardiac energy metabolism and reduced lipid peroxidation. PMID- 10381187 TI - Aluminum-induced oxidative events in cell lines: glioma are more responsive than neuroblastoma. AB - Aluminum, a trivalent cation unable to undergo redox reactions, has been linked to many diseases such as dialysis dementia and microcytic anemia without iron deficiency. It has also been implicated in Alzheimer's disease although this is controversial. Because cell death due to oxidative injury is suspected to be a contributory factor in many neurological diseases and aluminum neurotoxicity, glioma (C-6) and neuroblastoma (NBP2) cells were utilized to assess early changes in oxidative parameters consequent to a 48-h exposure to aluminum sulfate. A 500 microM concentration of this salt produced a significant increase in reactive oxygen species (ROS) production and a significant decrease in glutathione (GSH) content in glioma cells. However, the same concentration of the aluminum salt did not lead to any significant changes in the neuroblastoma cells. Mitochondrial respiratory activity in glioma cells was also found to be significantly higher in the aluminum treated cells. As judged by morin-metal complex formation, aluminum can enter glioma cells much more readily than neuroblastoma cells. Thus, it is possible that the cerebral target following an acute exposure to aluminum may be glial rather than neuronal. PMID- 10381188 TI - Ascorbic acid maintenance in HaCaT cells prevents radical formation and apoptosis by UV-B. AB - We have investigated the enzymatic reduction and accumulation of vitamin C in HaCaT epithelial cells. The subcellular localization and the activities of ascorbyl free radical reductase and dehydroascorbate reductase showed that mitochondrial, microsomal and plasma membranes fractions express high levels of ascorbyl free radical reductase activity, whereas dehydroascorbate reductase activity was found at low levels only in the post microsomal supernatant. We have also investigated cell proliferation and vitamin C accumulation induced by ascorbic acid 2-phosphate. This derivative caused no inhibition of cell growth, was uptaken from the extracellular medium and accumulated as ascorbic acid in mM concentrations. These results show that HaCaT cells possess very efficient systems to maintain high levels of both intracellular and extracellular ascorbic acid. The regeneration and uptake of ascorbic acid from extracellular medium contributes to the intracellular antioxidant capacity, as evaluated by 2',7' dihydrodichlorofluorescein staining. Consequently, cells became more resistant to free radical generation and cell death induced by UV-B irradiation. PMID- 10381189 TI - Localization of 4-hydroxy-2-nonenal-modified proteins in kidney following iron overload. AB - Intraperitoneal (IP) injection of ferric nitrilotriacetate (Fe-NTA) to rats and mice results in iron-induced free radical injury and cancer in kidneys. We sought to clarify the exact localization of acute oxidative damage in Fe-NTA-induced nephrotoxicity by performing immunogold light and electron microscopic (EM) techniques using an antibody against 4-hydroxy-2-nonenal (HNE)-modified proteins. Biochemical assays were done to provide complementary quantitative data. Renal accumulation of lipid peroxidation-derived aldehydes, such as malondialdehyde (MDA) and 4-hydroxy-2-alkenals (4-HDA), increased in parallel with protein carbonyl content, an indicator of protein oxidation, 30 min after administration of Fe-NTA. Immunogold light microscopy showed that HNE-modified proteins increased at 30 min with positivity localized to proximal tubular cells. Immunogold EM demonstrated that HNE-modified proteins were mainly in the mitochondria and nuclei of the proximal tubular epithelium. The intensity of labeling at both the light and EM levels increased together with levels of biochemically measured lipid peroxidation products and protein carbonyl content. Our data suggest that the mechanism of acute nephrotoxicity of Fe-NTA involves mitochondrial and nuclear oxidative damage, findings that may help to define the mechanisms of iron-induced cell injury. PMID- 10381190 TI - Intestinal damage induced by zinc deficiency is associated with enhanced CuZn superoxide dismutase activity in rats: effect of dexamethasone or thyroxine treatment. AB - Zinc has a wide spectrum of biological activities and its deficiency has been related to various tissue dysfunctions and alterations of normal cell metabolism. Zinc also plays an important role in the antioxidant cellular defenses being a structural element of the non-mitochondrial form of the enzyme superoxide dismutase (CuZnSOD). We have already reported that Zn deficiency induces severe alterations in the rat intestine, that are reverted by treatment with dexamethasone (Dex) or thyroxine (T4). Here we report a paradoxical increase of CuZnSOD activity in rat intestine after 20 and 40 days of zinc deficiency. The increase of CuZnSOD activity is not due to an upregulation of gene expression because both Northern and Western blot analysis indicate that CuZnSOD mRNA and protein levels are not affected by zinc deficiency. A significant increase of lipid peroxidation was also observed in duodenum and jejunum associated with zinc deficiency. Treatment with either Dex or T4 to zinc-deficient rats protects against intestinal oxidative damage and results in SOD activity similar to control rats. Because glutathione peroxidase and catalase activities decreased in zinc deficiency, we speculate that the increase in SOD activity may be associated with an accumulation of hydrogen peroxide that may activate inflammatory molecules, further worsening tissue damage. PMID- 10381191 TI - Singlet oxygen quenching and the redox properties of hydroxycinnamic acids. AB - The singlet oxygen quenching rate constants (kq) for a range of hydroxycinnamic acids in acetonitrile and D2O solutions were measured using time resolved near infrared phosphorescence in order to establish their antioxidant activity. The magnitude of kq observed depends on both the nature of the substituent groups and solvent polarity. The variations in kq depend on the energy of the hydroxycinnamic acid/molecular oxygen charge transfer states, (O2delta- ...HCAdelta+). In D2O the values of kq range from 4x10(7) M(-1) s(-1) to 4x10(6) M(-1) s(-1) for caffeic acid and o-coumaric acid respectively. In acetonitrile, the charge transfer energy levels are raised and this is reflected in lower singlet oxygen quenching rate constants with a kq value of 5x10(6) M(-1) s(-1) for caffeic acid. The phenoxyl radical spectra derived from the hydroxycinnamic acids were determined using pulse radiolysis of aqueous solutions and the reduction potentials were found to range from 534 to 596 mV. A linear correlation is observed between reduction potential, and hence free energy for electron transfer, and log kq. These correlations suggest a charge transfer mechanism for the quenching of singlet oxygen by the hydroxycinnamic acids. PMID- 10381192 TI - Noninvasive evaluation of in vivo free radical reactions catalyzed by iron using in vivo ESR spectroscopy. AB - The noninvasive, real time technique of in vivo electron spin resonance (ESR) spectroscopy was used to evaluate free radical reactions catalyzed by iron in living mice. The spectra and signal decay of a nitroxyl probe, carbamoyl-PROXYL, were observed in the upper abdomen of mice. The signal decay was significantly enhanced in mice subcutaneously loaded with ferric citrate (0.2 micromol/g body wt) and the enhancement was suppressed by pre-treatment with either desferrioxamine (DF) or the chain breaking antioxidant Trolox, but only slightly suppressed by the hydroxyl radical scavenger DMSO. To determine the catalytic form of iron, DF was administered at different times with respect to iron loading: before, simultaneously, and after 20 and 50 min. The effect of DF on signal decay, liver iron content, iron excretion, and lipid peroxidation (TBARs) depended on the time of the treatment. There was a good correlation between the signal decay, iron content, and lipid peroxidation, indicating that "chelatable iron" contributed to the enhanced signal decay. The nitroxyl probe also exhibited in vivo antioxidant activity, implying that the process responsible for the signal decay of the nitroxyl probe is involved in free radical oxidative stress reactions catalyzed by iron. PMID- 10381193 TI - Oxidative inactivation of human and sheep platelet membrane-associated phosphotyrosine phosphatase activity. AB - Incubation of human or sheep platelet crude membranes with xanthine oxidase/hypoxanthine in the presence of Fe2+/ADP inactivated phosphotyrosine phosphatase (PTPase, protein-tyrosine-phosphate-phosphohydrolase, EC 3.1.3.48) activity in a time-dependent manner, this inhibition being significant within 5 min of treatment. The dynamics of protein thiols differed depending on the platelet species, but in any case decreases in protein thiols were only visible 20-45 min after the start of the treatment. The inhibition of PTPase activity in general showed good a correlation with the production of thiobarbituric acid reactive substances (TBARS). The results with several antioxidants suggest that the inhibition of PTPase activity is related to the generation of alkoxyl and/or peroxyl radicals. Furthermore, the formation of fluorescent products and changes in amino groups were observed only after long incubation times with the oxidizing agents, these fluorescent products and the residual enzyme activity remaining in the membrane fraction. Treatment of platelet membranes with trans-2-nonenal and n heptaldehyde, but not with acetaldehyde, also inhibited membrane-associated PTPase activity. However, the amount of protein thiols was reduced only by treatment with trans-2-nonenal. Fluorescence product formation was always higher with trans-2-nonenal, these products being mainly located in the protein fraction. The results with aldehydes suggest that secondary degraded products of lipid hydroperoxides affect PTPase activity. Kinetic studies of PTPase activity indicated that with all treatments enzyme inhibition is mainly due to a decrease in the Vmax value. The results of fluorescence anisotropy measurements of labeled platelet membranes did not support the notion of a contribution of the lipid organization to peroxidation-mediated PTPase inhibition. All the above results indicate that platelet membrane-associated PTPase inhibition due to treatment with xanthine oxidase/ hypoxanthine in the presence of Fe2+/ADP is a very complex, time-dependent process, and that it is probably related, at least after long periods of peroxidation, to changes in protein thiols and amino groups. We predict that the sensitivity of PTPase to lipid peroxidation must be physiologically relevant because of the increasing importance of tyrosine phosphorylation in signal transduction, in general, and in platelet activation and aggregation in particular. PMID- 10381194 TI - Antioxidant activity applying an improved ABTS radical cation decolorization assay. AB - A method for the screening of antioxidant activity is reported as a decolorization assay applicable to both lipophilic and hydrophilic antioxidants, including flavonoids, hydroxycinnamates, carotenoids, and plasma antioxidants. The pre-formed radical monocation of 2,2'-azinobis-(3-ethylbenzothiazoline-6 sulfonic acid) (ABTS*+) is generated by oxidation of ABTS with potassium persulfate and is reduced in the presence of such hydrogen-donating antioxidants. The influences of both the concentration of antioxidant and duration of reaction on the inhibition of the radical cation absorption are taken into account when determining the antioxidant activity. This assay clearly improves the original TEAC assay (the ferryl myoglobin/ABTS assay) for the determination of antioxidant activity in a number of ways. First, the chemistry involves the direct generation of the ABTS radical monocation with no involvement of an intermediary radical. Second, it is a decolorization assay; thus the radical cation is pre-formed prior to addition of antioxidant test systems, rather than the generation of the radical taking place continually in the presence of the antioxidant. Hence the results obtained with the improved system may not always be directly comparable with those obtained using the original TEAC assay. Third, it is applicable to both aqueous and lipophilic systems. PMID- 10381195 TI - Dietary supplementation with beta-carotene, but not with lycopene, inhibits endothelial cell-mediated oxidation of low-density lipoprotein. AB - Carotenoids may protect low-density lipoprotein from oxidation, a process implicated in the development of atherosclerosis. Our previous studies showed that in vitro enrichment of low-density lipoprotein (LDL) with beta-carotene protected it from cell-mediated oxidation. However, in vitro enrichment with either lutein or lycopene actually enhanced oxidation of the LDL. In the present studies we have examined the impact of LDL carotenoid content on its oxidation by human aortic endothelial cells (EaHy-1) in culture, comparing the effects of in vivo supplementation with in vitro enrichments. The beta-carotene content in human LDL was increased three- to sixfold by daily supplementation with 15 mg beta-carotene for 4 weeks, and the lycopene content of LDL in other individuals was increased two- to threefold by ingestion of one glass (12 ounce) of tomato juice daily for 3 weeks. LDL isolated from these healthy, normolipidemic donors not taking supplemental carotenoid was incubated at 0.25 mg protein/ml with EaHy 1 cells in Ham's F-10 medium for up to 48 h. Following dietary beta-carotene supplementation, LDL oxidation (as assessed by formation of lipid hydroperoxides) was markedly inhibited, to an even greater extent than was observed for LDL enriched in vitro with beta-carotene (that resulted in an 11- to 12-fold increase in LDL beta-carotene). No effect on cell-mediated oxidation was observed, however, for LDL enriched in vivo with lycopene. Thus, beta-carotene appears to function as an antioxidant in protecting LDL from cell-mediated oxidation although lycopene does not. The fact that the three- to sixfold enrichments of LDL with beta-carotene achieved by dietary supplementation were more effective in inhibiting oxidation than the 11- to 12-fold enrichments achieved by an in vitro method suggests that dietary supplementation is a more appropriate procedure for studies involving the enrichment of lipoprotein with carotenoids. PMID- 10381196 TI - Kinetics of superoxide-induced exchange among nitroxide antioxidants and their oxidized and reduced forms. AB - Nitroxide stable radicals generally serve for probing molecular motion in membranes and whole cells, transmembrane potential, intracellular oxygen and pH, and are tested as contrast agents for magnetic resonance imaging. Recently nitroxides were found to protect against oxidative stress. Unlike most low molecular weight antioxidants (LMWA) which are depleted while attenuating oxidative damage, nitroxides can be recycled. In many cases the antioxidative activity of nitroxides is associated with switching between their oxidized and reduced forms. In the present work, superoxide radicals were generated either radiolytically or enzymatically using hypoxanthine/xanthine oxidase. Electron paramagnetic resonance (EPR) spectrometry was used to follow the exchange between the nitroxide radical and its reduced form; whereas, pulse radiolysis was employed to study the kinetics of hydroxylamine oxidation. The results indicate that: a) The rate constant of superoxide reaction with cyclic hydroxylamines is pH-independent and is lower by several orders of magnitude than the rate constant of superoxide reaction with nitroxides; b) The oxidation of hydroxylamine by superoxide is primarily responsible for the non-enzymatic recycling of nitroxides; c) The rate of nitroxides restoration decreases as the pH decreases because nitroxides remove superoxide more efficiently than is hydroxylamine oxidation; d) The hydroxylamine reaction with oxidized nitroxide (comproportionation) might participate in the exchange among the three oxidation states of nitroxide. However, simulation of the time-dependence and pH-dependence of the exchange suggests that such a comproportionation is too slow to affect the rate of non-enzymatic nitroxide restoration. We conclude that the protective activity of nitroxides in vitro can be distinguished from that of common LMWA due to hydroxylamine oxidation by superoxide, which allows nitroxide recycling and enables its catalytic activity. PMID- 10381197 TI - Salicylate promotes myeloperoxidase-initiated LDL oxidation: antagonization by its metabolite gentisic acid. AB - The oxidative modification of low density lipoprotein (LDL) may play a significant role in atherogenesis. Tyrosyl radicals generated by myeloperoxidase (MPO) can act as prooxidants of LDL oxidation. Taking into consideration, that monophenolic compounds are able to form phenoxyl radicals in presence of peroxidases, we have tested salicylate, in its ability to act as a prooxidant in the MPO system. Measurement of conjugated dienes and lipid hydroperoxides were taken as indicators of lipid oxidation. Exposure of LDL preparations to MPO in presence of salicylate revealed that the drug could act as a catalyst of lipid oxidation in LDL. The radical scavenger ascorbic acid as well as heme poisons (cyanide, azide) and catalase were inhibitory. The main metabolite of salicylic acid, gentisic acid, showed inhibitory action in the MPO system. Even when lipid oxidation was maximally stimulated by salicylate the LDL oxidation was efficaciously counteracted in presence of gentisic acid at salicylate/gentisic acid ratios that could be reached in plasma of patients receiving aspirin medication. Gentisic acid was also able to impair the tyrosyl radical catalyzed LDL peroxidation. The results suggest that salicylate could act like tyrosine via a phenoxyl radical as a catalyst of LDL oxidative modification by MPO. But the prooxidant activity of this radical species is effectively counteracted by the salicylate metabolite gentisic acid. PMID- 10381198 TI - Molecular orbital theory applied to the study of nonsteroidal anti-inflammatory drug efficiency. AB - Using a simple quantum mechanical method, we calculated the energy of the highest occupied molecular orbital (E(HOMO)) of three groups of anti-inflammatory compounds, and we have found correlations between E(HOMO) of these molecules and experimental data previously reported on (1) inhibition of sheep-vesicular-gland prostaglandin cyclooxygenase by phenolic compounds, (2) inhibition of prostaglandin cyclooxygenase in mouse macrophages by salicylates, benzoates and phenols, and (3) peroxyl-radical scavenging and radioprotection of a bacterial virus by NSAID drugs, including metiazinic acid, sulindac, D-penicillamine, piroxicam, indomethacin, benoxaprofen, and aspirin. Our correlations using a systematic evaluation of the HOMO energies can be of predictive value in the search for new anti-inflammatory drugs as well as for new radioprotectors. PMID- 10381199 TI - Immunochemical detection of glyoxal DNA damage. AB - The relevance of reactive oxygen species (ROS) in the pathogenesis of inflammatory diseases is widely documented. Immunochemical detection of ROS DNA adducts has been developed, however, recognition of glyoxal-DNA adducts has not previously been described. We have generated a polyclonal antibody that has shown increased antibody binding to ROS-modified DNA in comparison to native DNA. In addition, dose-dependent antibody binding to DNA modified with ascorbate alone was shown, with significant inhibition by desferrioxamine, catalase, and ethanol. Minimal inhibition was observed with uric acid, 1,10-phenanthroline and DMSO. However, antibody binding in the presence of EDTA increased 3500-fold. The involvement of hydrogen peroxide and hydroxyl radical in ascorbate-mediated DNA damage is consistent with ascorbate acting as a reducing agent for DNA-bound metal ions. Glyoxal is known to be formed during oxidation of ascorbate. Glyoxylated DNA, that previously had been proposed as a marker of oxidative damage, was recognised in a dose dependent manner using the antibody. We describe the potential use of our anti-ROS DNA antibody, that detects predominantly Fenton type mediated damage to DNA and report on its specificity for the recognition of glyoxal-DNA adducts. PMID- 10381201 TI - Damage to hemoglobin by radiation-generated serum albumin radicals. AB - We have studied the effects of the interaction of radiation generated human serum albumin radicals (HSA*) with human hemoglobin molecules (Hb). Diluted Hb aqueous solutions were irradiated under N2O or argon without HSA and in the presence of HSA. Analysis of Hb absorbance spectra in the visible range, cross-linking of HSA* radicals with Hb molecules and functional properties of Hb were investigated. The degree of Hb destruction estimated on the basis of changes in the absorption spectra indicated that the effectiveness of HSA* radicals generated under N2O for Hb destruction was approximately equal to that of *OH radicals. In this case mainly *OH radicals formed the secondary HSA* radicals. However, during the irradiation Hb + HSA under argon the presence of equivalent amounts of oxidizing and reducing products of water radiolysis lowers the degree of Hb destruction. Some reactions of HSA* radicals with Hb molecules lead to the formation of covalent bonds between the molecules of both proteins. The following types of hybrids could be distinguished: Hb monomer-HSA, Hb dimer-HSA and higher aggregates. Structural changes of Hb by HSA* radicals were reflected by alterations in the oxygen affinity (increase) and cooperativity (decrease) of Hb. The results obtained indicate that in the experimental systems studied, the HSA* radical reactions with Hb molecules are favoured over recombination reactions of HSA* radicals. On this basis one can suggest that in the studied systems Hb plays the role of an acceptor of radical energy located on HSA. PMID- 10381200 TI - Differential cytostatic effects of NO donors and NO producing cells. AB - A 3-h exposure to NO donors (spermine-NO, DETA-NO, or SNAP), or to NOS II expressing cells (activated macrophages or EMT6 cells) reversibly inhibited DNA synthesis in K562 tumor cells. In GSH-depleted K562 cells, cytostasis remained reversible when induced by DETA-NO or NOS II activity, but became irreversible after exposure to spermine-NO or SNAP. Only SNAP and spermine-NO efficiently inhibited GAPDH, an enzyme with a critical thiol, in GSH-depleted cells. Thus, the irreversible cytostasis induced in GSH-depleted cells by spermine-NO or SNAP can be tentatively attributed to S-nitrosating or oxidizing species derived from NO. However, these species did not contribute significantly to the early antiproliferative effects of macrophages. Ribonucleotide reductase, a key enzyme in DNA synthesis. has been shown to be inhibited by NO. Supplementation of the medium with deoxyribonucleosides to bypass RNR inhibition restored DNA synthesis in target cells exposed to DETA-NO and NO-producing cells, but was inefficient for GSH-depleted cells previously submitted to spermine-NO or SNAP. These cells also exhibited a persistent depletion of the dATP pool. In conclusion, GSH depletion reveals striking qualitative differences in the nature of the toxic effectors released by various NO sources, questioning the significance of S nitrosating or oxidizing nitrogen oxides in NOS II-dependent cytostasis. PMID- 10381202 TI - Antioxidants and mitochondrial respiration in lung, diaphragm, and locomotor muscles: effect of exercise. AB - Previous studies have shown that exhaustive exercise may increase reactive oxygen species (ROS) generation in oxidative muscles that may in turn impair mitochondrial respiration. Locomotor muscles have been extensively examined, but there is few report about diaphragm or lung. The later is a privileged site for oxygen transit. To compare the antioxidant defense system and mitochondrial function in lung, diaphragm and locomotor muscles after exercise, 24 young adult male rats were randomly assigned to a control (C) or exercise (E) group. E group rats performed an exhaustive running test on a motorized treadmill at 80-85% VO2max Mean exercise duration was 66+/-2.7 min. Lung, costal diaphragm, mixed gastrocnemius, and oxidative muscles (red gastrocnemius and soleus: RG/SOL homogenate) were sampled. Mitochondrial respiration was assessed in tissue homogenates by respiratory control index (RCI: rate of uncoupled respiration/rate of basal respiration) measurement. Lipid peroxidation was evaluated by malondialdehyde concentration (MDA) and we determined the activity of two antioxidant enzymes: superoxide dismutase (SOD) and glutathione peroxidase (GPX). We found elevated basal (C group data) SOD and GPX activities in both lung and diaphragm compared to locomotor muscles (p<.001). Exercise led to a rise in GPX activity in red locomotor muscles homogenate (GR/SOL; C = 10.3+/-0.29 and E = 14.4+/-1.51 micromol x min(-1) x gww(-1); p<.05), whereas there was no significant change in lung and diaphragm. MDA concentration and mitochondrial RCI values were not significantly changed after exercise. We conclude that lung and diaphragm had higher antioxidant protection than locomotor muscles. The exercise test did not lead to significant oxidative stress or alteration in mitochondrial respiration, suggesting that antioxidant function was adequate in both lung and diaphragm in the experimental condition. PMID- 10381203 TI - Effect of superoxide dismutase, catalase, chelating agents, and free radical scavengers on the toxicity of alloxan to isolated pancreatic islets in vitro. AB - The effect of superoxide dismutase, catalase, metal-chelating agents and hydroxyl radical scavengers on the toxicity of alloxan to isolated ob/ob mouse pancreatic islets in vitro has been compared with the reported ability of such substances to protect against alloxan diabetes in vivo. Superoxide dismutase and catalase protected beta-cells of isolated pancreatic islets against alloxan cytotoxicity, as did the hydroxyl radical scavengers dimethyl sulfoxide (DMSO) and butanol. However, 1,3-dimethylurea and thiourea, that are recognised as effective hydroxyl radical scavengers and that protect animals against the diabetogenic effects of alloxan, were without effect. Similarly, desferrioxamine, that inhibits hydroxyl radical formation from alloxan in chemically defined systems, did not protect against alloxan toxicity. Diethylenetriamine pentaacetic acid, which does not inhibit hydroxyl radical formation from alloxan, also gave no significant protection. The results indicate a role for superoxide radical and hydrogen peroxide in the mechanism of toxicity of alloxan but do not support the involvement of the hydroxyl radical in this process. Alternative explanations must be sought for the ability of hydroxyl radical scavengers and metal-chelating agents to protect against alloxan toxicity in vivo. PMID- 10381204 TI - Induction of DNA strand breakage and base oxidation by nitroxyl anion through hydroxyl radical production. AB - Nitroxyl anion (NO-), the one-electron reduction product of nitric oxide (NO*), has been reported to be formed under various physiological conditions and to be cytotoxic, although the mechanism responsible for the toxic effects has not been identified. We have studied the effects of NO- generated from Angeli's salt (sodium trioxodinitrate) or Piloty's acid (N-hydoxybenzenesulfonamide) on DNA strand breakage and DNA base oxidation in vitro. Induction of strand breakage was dose- and time-dependent upon incubation of plasmid pBR322 with Angeli's salt or Piloty's acid. Similarly, 8-oxo-2'-deoxyguanosine and malondialdehyde were formed when calf-thymus DNA or 2'-deoxyribose, respectively, were incubated with Angeli's salt. Electron acceptors (ferricyanide, 4-hydroxy-TEMPO), that convert NO to NO*, inhibited the reactions, indicating that NO , but not NO*, is responsible for the reactions. Furthermore, the reactions were also inhibited by the presence of hydroxyl radical (HO*) scavengers, antioxidants, metal chelators and superoxide dismutase and catalase, implying involvement of free HO*. These results suggest that NO- is a possible endogenous source of HO*, that may be formed either directly from the reaction product of NO- with NO* (N2O2*-) or indirectly through H2O2 formation. Thus NO may play an important role as a cause of diverse pathophysiological conditions such as inflammation and neurodegenerative diseases. PMID- 10381205 TI - Liver cancer is induced by a subnecrogenic dose of DENA when associated with fasting/refeeding: role of glutathione-transferase and lipid peroxidation. AB - In previous studies, we reported that fasting/refeeding has a role in sustaining the initiation of liver cancer by a subnecrogenic (noninitiating) dose of diethylnitrosamine (DENA). This research investigated whether the metabolic alterations imposed by fasting/refeeding provide an imbalance between the generation of carcinogenic molecules and the scavenger defense mechanisms in rat liver. Metabolism of DENA, levels of reduced glutathione (GSH) and GSH transferase (GST) activity, as well as basal and stimulated malondialdehyde (MDA) production, were examined. Rats fasted for 4 days showed a decrease in the liver levels of GSH, GST activity, monounsaturated fatty acids and % of labeled nuclei. After 1 day of refeeding, at which point DENA was administered, the levels of GSH recovered, GST activity remained below control values, basal and stimulated MDA production and content of total polyunsaturated fatty acids in liver phospholipids decreased. One day after DENA treatment, MDA production further decreased, although the % of labeled nuclei increased. No significant changes in the content of arachidonic acid, the main target of peroxidation, were observed at any time. The results indicated that the induction of the hepatocellular carcinoma was associated with a depression of GST activity and lipid peroxidation when rats were given 20 mg/kg of DENA after 1 day of refeeding after 4-day fasting. PMID- 10381206 TI - Inhibition of GTP binding to Rac2 by peroxynitrite: potential role for tyrosine modification. AB - Peroxynitrite is a potent oxidant generated by the reaction of nitric oxide (*NO) and superoxide anion (O2*-), and both can be produced in inflammatory tissues. In the present studies, we analyzed the effects of peroxynitrite treatment on the GTP-binding activity of Rac2, a low molecular weight GTP-binding protein important in regulating a number of cellular functions. Using a fluorescent analog of GTP (methylanthraniloyl guanosine triphosphate or mant-GTP) as a reporter group, we found that treatment of Rac2 with peroxynitrite inhibited the binding of mant-GTP to Rac2 in a dose-dependent manner. Peroxynitrite was also able to react directly with free mant-GTP, resulting in a significant decrease in mant-GTP fluorescence; however, the mechanism of peroxynitrite-mediated damage to mant-GTP was different than with Rac2. In the case of mant-GTP, protection from peroxynitrite-mediated oxidation was observed in the presence of the free radical scavengers, mannitol and DMTU. In contrast, DMTU was unable to prevent peroxynitrite-mediated inhibition of mant-GTP binding to Rac2. Instead, our data demonstrates a role for peroxynitrite-mediated tyrosine modification in the inhibition of mant-GTP binding to Rac2, and we were able to demonstrate the formation of a significant level of nitrotyrosine formation in Rac2 exposed to peroxynitrite. Thus, our studies support the premise that oxidative modification of key cellular proteins, such as Rac2, plays an important role in the cytotoxic effects observed for peroxynitrite and other reactive oxidants. PMID- 10381207 TI - Expression of human FALS SOD in motorneurons of Drosophila. AB - Mutations in human CuZn superoxide dismutase (SOD) have been associated with familial amyotrophic lateral sclerosis (FALS). Although leading to many experimental advances, this finding has not yet led to a clear understanding of the biochemical mechanism by which mutations in SOD promote the degeneration of motorneurons that causes this incurable paralytic disease. To explore the biochemical mechanism of FALS SOD-mediated neuropathogenesis, we used transgenic methodology to target the expression of a human FALS SOD to motorneurons of Drosophila, an organism known for its phenotypic sensitivity to genetic manipulation of SOD. Earlier, we showed that targeted expression of human SOD in motorneurons of Drosophila causes a dramatic extension of adult lifespan (>40%) and rescues most of the phenotypes of SOD-null mutants. Using the same genetic system, we now ask if targeted expression of a mutant allele of human SOD that is associated with FALS causes paralysis and premature death, or is otherwise injurious in Drosophila as it is in humans and transgenic mice. Here we report that high-level expression of a human FALS SOD in motorneurons is not detrimental and does not promote paralysis and premature death when expressed in motorneurons of Drosophila. In sharp contrast, the expression of FALS SOD in Drosophila actually extends lifespan, augments resistance to oxidative stress and partially rescues SOD-null mutants in a manner predicted by our earlier studies on the expression of wildtype human SOD in Drosophila motorneurons. PMID- 10381208 TI - New sensitive agents for detecting singlet oxygen by electron spin resonance spectroscopy. AB - Free radicals are well-established transient intermediates in chemical and biological processes. Singlet oxygen, though not a free radical, is also a fairly common reactive chemical species. It is rare that singlet oxygen is studied with the electron spin resonance (ESR) technique in biological systems, because there are few suitable detecting agents. We have recently researched some semiquinone radicals. Specifically, our focus has been on bipyrazole derivatives, which slowly convert to semiquinone radicals in DMSO solution in the presence of potassium tert-butoxide and oxygen. These bipyrazole derivatives are dimers of 3 methyl-1-phenyl-2-pyrazolin-5-one and have anti-ischemic activities and free radical scavenging properties. In this work, we synthesized a new bipyrazole derivative, 4,4'-bis(1p-carboxyphenyl-3-methyl-5-hydroxyl)-pyrazole, DRD156. The resulting semiquinone radical, formed by reaction with singlet oxygen, was characterized by ESR spectroscopy. DRD156 gave no ESR signals from hydroxyl radical, superoxide, and hydrogen peroxide. DRD156, though, gives an ESR response with hypochlorite. This agent, nevertheless, has a much higher ability to detect singlet oxygen than traditional agents with the ESR technique. PMID- 10381209 TI - Neuroinflammatory processes are important in neurodegenerative diseases: an hypothesis to explain the increased formation of reactive oxygen and nitrogen species as major factors involved in neurodegenerative disease development. AB - The hypothesis, as stated in the title, has arisen from the failure of simpler notions to explain a series of otherwise difficult to understand observations and the mounting evidence, in a broader sense, that inflammatory processes in the CNS are important etiologically in neurodegenerative diseases. Novel aspects include the primacy of inflammatory processes, within the CNS, which leads to increased formation of "proinflammatory" cytokines that lead to increased formation of reactive oxygen species (ROS) and mediation of the upregulation of genes that produce toxic products such as reactive nitrogen species (RNS). Here I utilize important background reports and synthesize ideas to help account for the noted increases in ROS and RNS and their biological reaction products in neurodegenerative diseases. The uniqueness of the CNS inflammatory processes include minimal damping of amplification processes, such as proinflammatory cytokine-mediated cascades, combined with unique genetic defects, that act in combination with other risk factors to repeatedly "spark" the inflammatory cascades to account for some of the major differences in neurodegenerative diseases. This hypothesis can be experimentally examined by development of definitive methods to quantitate unique products that are formed by processes predicted to occur under neurodegenerative conditions. PMID- 10381210 TI - Safety profile of meropenem: a review of nearly 5,000 patients treated with meropenem. AB - Meropenem is a parenteral carbapenem that has been used clinically since 1994. Since the first review of its safety profile in 1995, the patient database has increased substantially. This new safety analysis includes data from 46 clinical trials in hospitalized patients with serious bacterial infections. The additional data comprise patients with lower respiratory tract and intra-abdominal infections, septicaemia and meningitis, and cancer patients with febrile neutropenia, and represents a group of more severely ill patients compared with the earlier review. In total, 4872 patients with 5026 meropenem treatment exposures were compared with 4642 patients treated with comparator agents (4752 exposures). Meropenem was administered most often by intravenous injection at 1g or 500 mg every 8 h. Meropenem-related adverse events most frequently reported were diarrhoea (2.3%), rash (1.4%), nausea/vomiting (1.4%) and injection site inflammation (1.1%). The most commonly reported meropenem-related laboratory adverse events were thrombocytosis (1.6%) and increased hepatic enzymes (1.5 4.3%). In meropenem-treated patients with meningitis, the incidence of seizures was low and none were drug related. In patients with infections other than meningitis, the incidence of seizures considered by the investigators to be related to meropenem was 0.08%. In general, the safety profile of meropenem was similar to that of the comparator agents. Withdrawals and deaths were similarly infrequent in the meropenem, cephalosporin and imipenem-cilastatin groups. Increased doses of meropenem were not associated with an increased incidence of adverse events. Meropenem was well tolerated in all patients, including children and patients with neutropenia. This new analysis supports the previous findings that meropenem has a favourable and acceptable safety profile. PMID- 10381212 TI - Reduced passive measles immunity in infants of mothers who have not been exposed to measles outbreaks. AB - Natural measles infection usually confers life-long immunity which is transferred from mothers to their offspring, protecting them from natural measles until the age of about 12 months. Recently, however, natural measles has been observed with increased frequency in infants under the age of 12 months. Natural measles outbreaks in the city of Sapporo have been suppressed by widely applied measles vaccination. Passive measles immunity in 160 neonates (cord blood), born during the last 17 y in Sapporo, Japan was determined by a neutralization (NT) antibody test. The mothers of these infants had had natural measles infection during childhood. Geometric mean titres (GMTs) of cord blood NT antibodies gradually decreased after 1989 and the GMTs of the most recently born infants were significantly lower than those of infants born in the first few years of the study. These observations suggest that even in mothers who experienced natural measles in childhood, recurrent exposure to natural measles is necessary in order to maintain adequate antibody levels for effective passive immunity of their infants. PMID- 10381211 TI - Clinical and laboratory findings in immunocompetent patients with persistent parvovirus B19 DNA in bone marrow. AB - The clinical relevance of parvovirus B19 DNA persistence in bone marrow was examined in 10 immunocompetent individuals undergoing examinations for unexplained fever, arthralgia or chronic leukopenia. Common causes of these symptoms had been ruled out and bone marrow aspiration was indicated at this stage of investigation. In addition to morphological analysis of the bone marrow, a test for B19 DNA was performed with 2 nested PCRs. Five of these 10 selected patients had detectable B19 DNA in their bone marrow, whereas no viraemia was observed. Additional bone marrow samples were collected at least 6 months after the first sample from the B19 DNA-positive patients, of whom 3 were found to be still positive. Indeed, 2 of the patients have been positive for more than 5 y of follow-up. Sera from all patients with persistent B19 DNA in bone marrow could neutralize the virus. One patient responded to treatment with immunoglobulin but later relapsed. No other cause of the symptoms was found, despite extensive investigations, and at least some of the prolonged disease manifestations may be due to parvovirus B19. PMID- 10381213 TI - TT virus infection among IVDUs in south western China. AB - To elucidate the prevalence and clinical implications of infection with the newly described TT virus (TTV) among intravenous drug users (IVDUs) in Yunnan, southwest China, serum samples from 158 IVDUs (129 M, 29 F; mean age 26.1 +/- 5.5 years) were examined for TTV DNA by semi-nested polymerase chain reaction (PCR) using primers derived from the open reading frame (ORFI) of TTV DNA. The seroprevalence of viral markers of HIV, HBV, HCV and GB virus C/hepatitis G virus (GBV-C) infection was also examined. A molecular evolutionary analysis was performed. Thirty one (20%) of the IVDUs were positive for TTV DNA, and 34 (22%), 6 (4%), 98 (62%), 76 (48%), 136 (86%) and 65 (41%) were positive for anti-HIV, HBsAg, anti-HBs, anti-HBc, anti-HCV and GBV-C RNA, respectively. When all the subjects were classified according to TTV DNA positivity, no significant differences were observed in demographic, biochemical or virological characteristics between the 2 groups. TTV infection was in all cases associated with co-infection with 1 or more of the other aforementioned viruses. There were no significant differences in the various combinations of co-infection between TTV positive and negative groups. Phylogenetic analysis revealed that the TTV isolates obtained in the study could be grouped mainly into TTV genotype 1, and that some of the isolates belonged to subgroups other than those previously described. These results indicate that: 1) TTV infection is prevalent among IVDUs in China; 2) TTV probably has minor liver pathogenicity; and 3) new subgroups of genotype 1 and 2 exist in China. PMID- 10381214 TI - Mutations in the NS5A gene predict response to interferon therapy in Japanese patients with chronic hepatitis C and cirrhosis. AB - The virus genotype, serum HCV-RNA level and liver histology are reported to be important factors in the response to interferon therapy. Recent studies have revealed that HCV NS5A 2209-2248 amino acid changes affect the response to interferon therapy of genotype 1b chronic hepatitis C. In contrast, some studies done in western countries have reported no such correlation. In the present study, interferon therapy was given to 58 Japanese patients, including 15 liver cirrhosis patients. NS5A 2209-2248 changes, the serum HCV level, ALT level, age and histology were examined in relation to the interferon effect. Twenty-four of the 58 patients (41%) showed a sustained virological response to the therapy. The responses to interferon therapy were significantly correlated with NS5A 2209-2248 changes (p < 0.0001), the HCV-RNA level (p < 0.0001) and histology (p < 0.0060). Among 15 liver cirrhosis patients, 3 of 6 mutant type patients showed a sustained virological response; 5 intermediate and 4 with wild type virus infected patients showed no responses. In conclusion, NS5A 2209-2248 changes may be a useful predictive marker of response to interferon therapy in addition to the serum HCV RNA level even in histologically advanced patients. PMID- 10381215 TI - Prevalence of hepatitis C and G virus infections among intravenous drug users in Slovenia and Croatia. AB - The prevalence of hepatitis C virus (HCV) and hepatitis G virus (HGV) infections was assessed in 115 Slovenian and 102 Croatian intravenous drug users (IVDUs). HCV and HGV infections were detected in 60 (52.2%) and 61 (53.0%) Slovenian IVDUs and in 70 (68.6%) and 39 (38.2%) Croatian IVDUs, respectively. The established prevalence of both HCV and HGV infection in Croatian IVDUs are the lowest found to date among IVDUs. HCV positive Slovenian and Croatian IVDUs were significantly older and reported longer duration of the intravenous drug use in comparison with HCV negative IVDUs. In contrast, no significant differences in both parameters were found among HGV-positive and -negative IVDUs. PMID- 10381216 TI - Short-term risk for AIDS-indicator diseases predicted by plasma HIV-1 RNA and CD4+ lymphocytes. AB - The objective of this study was to assess the value of quantitative HIV-1 RNA as a predictor for the short-term risk of developing AIDS-defining events in comparison with CD4 cell counts. A total of 1,028 samples from 324 patients were analysed. Median initial CD4 cell counts and HIV-1 RNA were 249 x 10(6)/l (range 0-1400 x 10(6)/l) and 4.5 log copies/ml (range: 2.3-6.4 log copies/ml). CD4 cell counts and viral load (VL) values obtained the year before a single AIDS indicator disease were selected to define the risk of developing that event. Cox regression models with CD4 cell counts and VL values treated as time-dependent covariates were performed to analyse the risk for developing certain events. Receiver operating characteristic (ROC) curves were used to compare CD4 cell counts and VL values as predictive markers for progression. During a median follow-up of 870 d (range 30-1381 d), 132 patients developed AIDS. Median log VL values during the year before the event were 3.6 for non-progressors and 5.2 for those who developed AIDS (p < 0.0001). Minimum log VL threshold values for developing diseases were 2.3 for tuberculosis, 3.8 for Candida esophagitis, 4.4 for wasting syndrome, 4.5 for CMV disease and 4.7 for PCP. VL values were not, however, a better predictive marker for developing specific events than were CD4 cell counts. Although we have identified VL thresholds for the risk of developing certain AIDS-indicator diseases, the indication for starting prophylactic regimens may still be based on CD4 cell counts. PMID- 10381217 TI - Subsets of T-cells and in vitro cytokine production after measles and varicellae zoster virus antigen stimulation in allogeneic BMT patients. AB - This study was performed to analyse differences in T-cell proliferation induced by a latent virus, varicellae-zoster virus (VZV) and a non-latent virus, measles virus, in patients after allogeneic bone marrow transplantation (BMT). The lymphoproliferative response to measles antigen, VZV-antigen (VZV-ag), and phytohemagglutinin (PHA) was measured by 3H-thymidine incorporation, and interferon-gamma (IFN-gamma) and interleukin-10 (IL-10) analyses in supernatants after in vitro stimulation of peripheral blood mononuclear cells (PBMC) from 22 patients and 18 healthy controls. The cytokine levels were correlated with T-cell subsets by FACS analyses. At the antigen concentrations used, VZV-ag induced higher levels of IFN-gamma (p < 0.05) than did the measles antigen, whereas the levels of IL-10 were similar. Patients without a cell mediated immune (CMI) response to VZV-ag or measles antigen had lower CD4+ T-cell counts than did controls (p < 0.01 in both cases) and lower IFN-gamma production after non specific PHA stimulation (p <0.01). The IFN-gamma and IL-10 levels after measles antigen stimulation correlated with the number of CD4+ T-cells (p < 0.01 and p < 0.05, respectively), and after VZV-ag mainly to the number of CD8+ T-cells (p < 0.01 and p < 0.05, respectively). These results suggest that there is a difference in the types of T-cells that respond to VZV-ag and measles antigen stimulation, respectively. The impaired CMI response to viral antigens seen in many patients may be explained both by a low number of CD4+ T-cells and by a cell dysfunction. PMID- 10381218 TI - Serological evidence of Ehrlichia infection in Swedish Lyme borreliosis patients. AB - We studied sera from patients who had participated in a prospective study of borreliosis in Sweden and had acquired tick bites in areas of the country with a high prevalence of granulocytic ehrlichial infections in animals. The sera were examined for IgG anti Ehrlichia antibodies by an indirect immunofluorescence assay using a locally isolated bovine Ehrlichia antigen. Confirmation of the serological results was done at the Unite des Rickettsies, Marseille, France. Three out of 37 of the investigated patients and 1 out of 100 investigated healthy blood donors had significant antibody titres to granulocytotropic Ehrlichiae. No patient or blood donor had specific antibody titres to Ehrlichia chaffeensis. These data suggest that Scandinavian Ehrlichia species can infect and evoke immunological response in tick-exposed humans. PMID- 10381219 TI - Epidemiology of tuberculosis in HIV-infected patients in Denmark. AB - Denmark is an area of low incidence of HIV and tuberculosis (TB). The number of newly reported cases of HIV has been stable during the 1990s, whereas the number of TB cases has doubled in Denmark in the past decade, mainly due to immigration. However, among native Danes the incidence of TB has increased in the younger age groups, indicating more newly infected persons. This study was performed in order to assess the impact of the HIV epidemic and immigration on TB incidence among native Danes. The study was also designed to reveal transmission patterns of TB among HIV-positive patients. Data from HIV-TB co-infected patients identified in the national registers of TB and AIDS from 1992-95 were collected retrospectively from medical records. Restriction fragment length polymorphism (RFLP) analyses of TB isolates from co-infected patients were compared with all patterns registered in the nationwide Danish RFLP database (approximately 1,700 patients). Sixty seven co-infected patients were identified, 26 Danes and 41 immigrants, representing only 4% of all TB cases during the study period. Danish co-infected patients were part of a cluster, i.e. they had a RFLP-pattern identical to a pattern in the national RFLP database, more often than immigrants (83% vs. 45%, p < 0.005). In only 2 cases were co-infected Danes and immigrants part of the same cluster. Danish HIV-TB co-infected patients were more often intravenous drug users than were co-infected immigrants (p < 0.0005). In conclusion, we found no evidence to suggest that the increase in TB incidence among young Danes was caused by the HIV-epidemic or transmission from immigrants. TB among HIV-positive Danes is most often due to recent infection. The patients often belong to a subpopulation living in Copenhagen characterized by intravenous drug use. PMID- 10381220 TI - Generalized mycobacterium genavense infection in HIV-infected patients: detection of the mycobacterium in hospital tap water. AB - We describe 3 HIV-infected patients with disseminated M. genavense infection. The use of corticosteroids possibly favoured colonization and dissemination of atypical mycobacteria in these patients with low CD4 cell counts and may have masked symptoms of infection. The fact that these patients were treated with highly active antiretroviral therapy (HAART) together with antimycobacterial therapy may explain that 1 patient was free from mycobacteria 16 months after the end of specific treatment. Hospital tap water contained M. genavense at a concentration of >10 bacteria/l as examined by PCR. This species caused 12% of cases of non-tuberculous disseminated mycobacteriosis in HIV-infected patients at our hospital. PMID- 10381221 TI - Retrospective evaluation of exposure to P. falciparum using antibodies to circumsporozoite protein and to cultured P. falciparum antigens. AB - A Danish school class travelled under primitive circumstances in East Africa for 28 d. Four out of 18 persons had febrile illnesses interpreted as malaria during the journey. Retrospective serological testing for antibodies against the P. falciparum circumsporozoite protein indicated a transmission rate of more than 80% despite extensive protection against mosquito bites. The study demonstrates the use of serological evaluation of malaria transmission risk as well as retrospective immunodiagnosis of clinical malaria. Three of the travellers with febrile illnesses used self-medication with mefloquine, and in 2 of the cases a diagnosis of malaria was supported by a positive immunofluorescence assay for malaria antibodies. PMID- 10381223 TI - High frequency of gastrointestinal parasites in refugees and asylum seekers upon arrival in Sweden. AB - The results of routine screening for intestinal parasites in 1377 refugees and asylum seekers within 2 weeks of arrival in Sweden showed that protozoa, mainly Giardia intestinalis, were found in 235/1377 (17%) and helminths, mainly hookworms, in 264/1377 (19%). Intestinal parasites were more frequently recovered in refugees coming from South East Asia, Africa and Latin America (infection rates 48%, 43% and 42%, respectively) than in those from Eastern Europe (22%) and the Middle East (32%). Refugees who reported gastrointestinal symptoms were less often infected than those without symptoms (p < 0.001). Of the European refugees, 127 came from Bosnia. A high rate of hookworms was found in this group (15%), suggesting that hookworms may also be transmitted in temperate areas under special conditions. We thus identified relatively high rates of pathogens in all groups of refugees. Screening may therefore be recommended, though more for the benefit of refugees than for the prevention of further spread of the infections. PMID- 10381222 TI - Low frequency of complications in imported falciparum malaria: a review of 222 cases in south-eastern Norway. AB - We performed a retrospective study of 222 cases of falciparum malaria diagnosed in Oslo and Akerhus counties, Norway, from January 1988 to December 1997. Except for 12 cases, all had acquired the disease in sub-Saharan Africa. Sixty-four (28.8%) cases occurred in assumed non-immune individuals; of these, 41 (64.1%) were compliant to recommended antimalarial chemoprophylaxis. The mean time lag from first symptom to diagnosis (total diagnosis delay) was 4.6 d (median 3 d, range 0-30 d) and the mean time from presentation to diagnosis (doctor's delay) was 1.3 d (median 0 d, range 0-25 d). There were no fatal cases, and only 8 (3.6%) had a complicated course. The following factors were significantly associated with development of complicated disease: higher age, non-immunity combined with chemoprophylaxis non-compliance, prolonged doctor's delay and prolonged total diagnosis delay (p < or = 0.05). Our data suggest that complicated disease in imported falciparum malaria may largely be prevented by high chemoprophylaxis compliance rates in non-immune travellers and a high index of suspicion in physicians evaluating febrile travellers. PMID- 10381225 TI - A 10-year retrospective study of infective endocarditis at a university hospital with special regard to the timing of surgical evaluation in S. viridans endocarditis. AB - A total of 154 episodes of infective endocarditis (IE) in 149 patients were studied retrospectively with special regard to the major aetiological groups and the surgical evaluation. There were 136 episodes of native valve endocarditis (NVE) (88%) and 18 episodes of prosthetic valve endocarditis (PVE) (12%). Three major groups of NVE crystallized: Streptococcus viridans in 37 (27%), Staphylococcus aureus in 39 (29%) and culture negative IE in 28 (21%) episodes. In these groups surgery during the active phase was required in 41, 28 and 18%, respectively. At the operation myocardial abscess was found in as many as 7/15 cases with S. viridans, but in only in 3/11 cases with S. aureus and 1/5 cases with culture negative IE. The mean duration of preoperative antibiotic treatment was 34 d. This long period of unsuccessful pharmacotherapy, preceded by a mean of 47 d from start of symptoms to admission to hospital, has probably resulted in the high frequency of myocardial abscess in S. viridans NVE. Surgical evaluation should be considered when fever persists beyond 10 d of adequate treatment, even in the absence of clinically apparent complications. Among the PVE episodes, 11/18 were managed with pharmacological treatment alone. Uncomplicated PVE may thus often be successfully treated with antibiotics alone. PMID- 10381226 TI - The anti-HBs response after 2 different accelerated intradermal and intramuscular schemes for hepatitis B vaccination. AB - To study early seroconversion rates after hepatitis B vaccination intramuscular (i.m.) and low-dose intradermal (i.d.) vaccination was compared when given either according to the registered 0, 4, 8 weeks scheme (scheme A), or to an accelerated 0, 2, 6 weeks scheme (scheme B). Medical staff received either 2 microg i.d. or 20 microg i.m. of a recombinant hepatitis B vaccine, in a non-randomized open trial. Two weeks after the third dose i.m. vaccinees overall had significantly higher rates of protective anti-HBs levels (anti-HBs > or = 10 IU/I), (23/30, 77%) compared with i.d. vaccinees (75/166, 45%) (p < 0.001). We conclude that when rapid protection against hepatitis B virus (HBV) infection is desirable, such as for post-exposure prophylaxis, an accelerated low-dose i.d. vaccination schedule cannot be used. PMID- 10381224 TI - Evidence of 2 waves of Chlamydia pneumoniae infection in Gavle, Sweden, 1990-96. AB - Chlamydia pneumoniae is a common respiratory tract pathogen. The majority of adults have serological evidence of previous exposure. Most infections are probably asymptomatic or subclinical. Recent studies have implicated C. pneumoniae as a risk factor for the development of cardiovascular disease. It was therefore of interest to study new blood donors collected between the years of 1990 and 1996 for the purpose of delineating the epidemiological situation in the Gavle area of Sweden. Sera from all first time blood donors over a 7 y period were tested for IgG, IgA and IgM antibodies to C. pneumoniae with a microimmunofluorescence test (MIF). Donors were subjectively healthy individuals between 18 and 65 y of age, (913 M, 752 F). Exposure to C. pneumoniae, expressed in terms of specific IgG antibodies in titres of > 1/32, increased for men in 1990-92 and for women in 1990-93. There was a decrease the following year for both sexes, followed by another increase in 1994-95. IgG antibodies in titres of > or = 512, and IgA antibodies in titres of > or = 1/64, were increased in 1990 91 for men and in 1994-96 for both sexes. The prevalence of specific antibodies increased throughout the 7 y period except for women 1995-96. Men had higher antibody titres than women throughout the entire study period. The results indicate that two waves of largely subclinical infection occurred in our area over the years 1990-96. PMID- 10381227 TI - Perinatal Epstein Barr virus infection in a premature infant. AB - This is a case of an infant boy born at 28 weeks gestational age who presented on the 42nd day of life with hepatosplenomegaly, haemolytic anaemia, thrombocytopenia and atypical lymphocytes on the peripheral blood smear. He had an Epstein Barr virus (EBV) viral capsid antigen (VCA) IgM antibody titre of 1:160 and a positive test for heterophil antibodies. The cytomegalovirus (CMV) IgM titre was 0.600 and CMV IgG 70 Au/ml. The infant died 10 d later and the autopsy showed CMV inclusion bodies in the lungs, liver and kidneys. EBV infection acquired perinatally, probably co-existing with CMV, may have led to a fatal disease. PMID- 10381228 TI - Primary sternal osteomyelitis and septicaemia due to Staphylococcus aureus. AB - Primary sternal osteomyelitis is rare in these recent decades. Only scattered cases have been reported, most of them in intravenous drug users. We report the case of an 88-y-old woman who presented a primary sternal infection due to Staphylococcus aureus associated with secondary septicaemia. The only predisposing factor was radiotherapy for a malignant tumour of the right mammary gland 20 y ago. Diagnostic evaluation and therapeutic management are briefly discussed. PMID- 10381229 TI - Group G streptococcus sacroilitis with sepsis in a 15-y-old adolescent. AB - Group G streptococci cause invasive infections of different tissues. Most infected patients have underlying diseases and are of adult age. Invasive group G streptococcal infections rarely occur in childhood and adolescence. A 15-y-old boy with a beta-haemolytic group G streptococcus sacroiliitis, sepsis and secondary pulmonary manifestations resembling an acute respiratory distress syndrome is described. PMID- 10381230 TI - Seroprevalence of Chlamydia pneumoniae in Chile. AB - We used microimmunofluorescence to survey the prevalence of antibodies to Chlamydia pneumoniae in 403 serum samples from asymptomatic subjects aged 6 months to 89 y in Santiago, Chile. The results suggest that Chlamydia pneumoniae infection is endemic in Chile, with a seroprevalence of 60% which does not differ by gender. PMID- 10381231 TI - A comparative study of infrared tympanic thermometry and rectal mercury thermometry. AB - In this study we compared infrared tympanic thermometry with rectal mercury thermometry and digital rectal thermometry in patients admitted to a medical department. We found that infrared tympanic thermometry has a low sensitivity for detecting fever. Digital rectal thermometry is a good alternative to rectal mercury thermometry. PMID- 10381232 TI - Activity limitation and chronic conditions in Canada's elderly, 1986-2011. AB - Together with all other developed countries, Canada's population is experiencing a significant increase in the proportion that is elderly. This paper examines basic linkages between individual ageing, the prevalence of various chronic health conditions, functional limitation and the receipt of help in activities of daily living (ADL) and instrumental activities of daily living (IADL) for the Canadian population using recent data from the National Population Health Survey (NPHS) as well as the Health and Activity Limitation Surveys (HALS) and the two General Social Surveys (GSS) with health data. Presented are age- and sex specific prevalence of chronic conditions and logistic regression is used to assess the impacts of different chronic conditions on the receipt of help for IADL and ADL. The importance of gender and living alone in influencing the receipt of help and also of use of formal agencies is presented using additional data from HALS. Findings from these analyses are also used to project changes in the distribution of health status defined by disability and receipt of help with IADL/ADL and, secondarily, by chronic condition. These analyses imply increases in demand for a range of health related services which will be 50 to 100% greater than the growth in the total elderly population. PMID- 10381233 TI - Gains in health expectancy from the elimination of diseases among older people. AB - PURPOSE: This paper examines a health expectancy based approach to obtaining disease-specific measures of the contribution of health problems to loss of healthy life among older people. Health expectancies combine mortality and morbidity into a single population health measure. The objectives of this study are to evaluate the usefulness of potential gains in health expectancies as a measure of health impact of various chronic diseases and injury among older people and to examine whether elimination of specific diseases and injuries leads to a compression or expansion of morbidity. Results are presented for Australians aged 65 years and over in 1993. RESULTS: The results highlight the importance of the chronic non-fatal diseases such as osteoarthritis and eyesight and hearing problems as causes of disability and handicap in older people. Elimination of such diseases results in an increase in healthy years of life while total life expectancy remains unchanged, leading to an absolute compression of morbidity. At the other extreme, elimination of highly fatal diseases such as cancer can result not only in an increase in healthy years but an even larger increase in years with disability, resulting in a relative expansion of morbidity. PMID- 10381234 TI - Receptivity to new technology among older adults. AB - PURPOSE: Both absolute and relative increases in the older adult population are occurring concurrently with the growth of high technology. Technological devices offer sophisticated solutions to some of the problems associated with ageing. This study borrows from the health utilization literature in order to develop and test a model for understanding receptivity to specific technological products by older adults. RESULTS: Receptivity is directly influenced by predispositional, need and social support factors, as well as by one's level of concern for problems that could be alleviated through the use of technology. Hierarchical regression equations reveal that this latter variable, concern, has the strongest influence on receptivity, while need factors display strong indirect effects. Those with unsatisfactory contact with others are also more receptive, suggesting that the lack of social support acts as a need factor. Contrary to past research, women are more receptive to technology than men. CONCLUSIONS: The results indicate that new technology geared toward enhancing the quality of life of seniors in their homes would be welcomed by many. PMID- 10381235 TI - Expectations of independence and life satisfaction among ageing spinal cord injured adults. AB - PURPOSE: The present study offers information about independence and life satisfaction over the lifespan for individuals with traumatic spinal cord injuries. METHODS: The study uses the health expectancy methodology to estimate expectations of the remaining years of life that may be spent in states of independence and satisfaction with life. SUBJECTS: The cohort studied had all incurred a spinal cord injury between the ages of 25 and 34, between the years 1945 and 1990 in central and south-eastern Ontario. RESULTS AND CONCLUSIONS: The study found that levels of independence and quality of life in the sample conformed closely to those found in other similar studies with the spinal cord injured population: 22% reported their own functional status as dependent, and 22% reported fair to poor life satisfaction. Expectations of independence appeared to decline steadily over the five decades studied, while expectations of modified independence increased proportionally. Estimates varied significantly for those with paraplegia vs. quadriplegia, and those with complete vs. incomplete lesions. Expectations of life satisfaction appeared to change after the 30 year mark; at that point, the balance changed so that expectations of dissatisfaction outweighed expectations of satisfaction. Multiple regression showed that independence was related to lesion level, completeness and recency of injury, and both independence and satisfaction were related to marriage and employment. PMID- 10381236 TI - Depression and life satisfaction among people ageing with post-polio and spinal cord injury. AB - PURPOSE AND BACKGROUND: Attention has recently begun to focus on the ageing of individuals with disability, not only as a long-term follow-up issue but as a unique developmental issue itself. The majority of individuals with an onset of disability before age 30 can now expect to live into their 60s, 70s and beyond. Most of the secondary medical conditions that foreshortened life expectancy have been controlled and improved rehabilitation techniques have evolved over the last 50 years. The average age of persons with post-polio in the United States is over 50 and the average age of persons with spinal cord injury is in the late 40s. New medical, functional and psychosocial problems have been discovered among persons ageing with these and other disabilities. Most of these problems lack sufficient scientific explanation, and therefore, clinical interventions. Quality of life (QOL) issues become involved as these changes occur. From a psychological perspective, QOL can be either positive, as reflected in high life satisfaction, or negative, as reflected in distress and depression. METHODS: This study reports on life satisfaction and depression in 360 persons, 121 with post-polio, 177 with SCI and 62 non-disabled age-matched comparisons. The Geriatric Depression Scale and the Older Adult Health and Mood Questionnaire assess depressive symptomatology and a 10-item life satisfaction scale with four-point ratings on each item used. RESULTS: Life satisfaction varied by the group, with the non disabled group higher than one or both of the other two groups on all scales and the post-polio group higher than the SCI group on six scales. Satisfaction with health, finances, work and overall life were most different. 22% of the post polio group, 41% of the SCI group and 15% of the non-disabled group had at least significant repressive symptomatology. CONCLUSION: The results for each group are discussed in terms of their relation to other coping variables that were assessed, particularly social support and coping methods. PMID- 10381237 TI - Ageing with spinal cord injury: the impact of spousal support. AB - PURPOSE: Research has offered ample evidence that spousal support can be seen as an important contributing factor to the ongoing health and well-being of ageing individuals, whether or not they have a spinal cord injury (SCI) or disability. In fact, spouses may be the most important element in successful rehabilitation and long-term home care for people with spinal cord injuries. This longitudinal study, which describes 225 British SCI long-term survivors, offers insight into marital status and its impact on general quality of life, depression, stress and community integration for individuals with SCI. RESULTS: The results demonstrate married individuals having less depression, greater life satisfaction and psychological well-being, and having better perceived quality of life. When controlling for age, duration of injury, and gender, marital status was a significant predictor of better perceived life satisfaction and quality of life. PMID- 10381238 TI - Disablement following stroke. AB - PURPOSE: Stroke is the most disabling chronic condition, newly affecting 35000 persons in Canada each year. Because of declining fatality, a growing number of persons will have to cope with stroke-related disability. The purpose of this paper is to describe the disabilities experienced by persons with stroke during the first year and explore the evolution of impairment, disability, handicap and health-related quality of life. SUBJECTS: The data for this paper come from a series of longitudinal and cross-sectional studies, collectively known as the McGill Stroke Rehabilitation Research Program. RESULTS: Within the first week post-stroke, getting out of bed and walking over a short distance, even with assistance, was a strong predictor of discharge home. Most of the improvement in measures of impairment and disability occurred during the first month and, by 3 months, there was still considerable room for improvement in all measures: 85% of persons were still impaired on gait speed, 78% had not reached age-specific norms for upper extremity function, 68% still demonstrated slow physical mobility, 37% needed some assistance with basic activities of daily living and 29% were still impaired on balance. By 1 year, 73% of persons scored the maximum for basic activities of daily living but 51 and 67% of persons reported their physical health and mental health to be lower than expected. Among a hardy group of stroke survivors, still living in the community 1 year post-stroke, the most striking area of difficulty was endurance, as measured by the 6 minute walk test. Those subjects well enough to complete this task (50% of sample) were able to walk, on average, only 250 metres, equivalent to 40% of their predicted ability. This series of snapshots taken at different points in time suggests that much of the improvement in impairment and disability occurs during the first month and then reaches a plateau. Handicap and quality of life continue to be issues later. Rehabilitation strategies need to consider the multifaceted nature of disablement, which in itself changes with time post-stroke. PMID- 10381239 TI - Energy medicine for long-term disabilities. AB - Energy medicine techniques derive from traditional Chinese medicine and are based upon the concept that health and healing are dependent upon a balance of vital energy, a still mind, and controlled emotions. Physical dysfunctions result from disordered patterns of energy of long standing and reversal of the physical problem requires a return to balanced and ordered energy. Qi Gong (Chi Kung) is a system which teaches an individual to live in a state of energy balance. Shen Qi is a sophisticated form of Qi Gong which relies on no external physical interventions but rather relies on mind control to prevent illness, heal existing physical and emotional problems, and promote health and happiness. This paper will describe the use of these techniques with people who have long-term physical disabilities. PMID- 10381240 TI - Residential transitions from nursing homes for adults with cerebral palsy. AB - PURPOSE: The impact of moving out of nursing homes into community-based settings for adults with cerebral palsy was assessed by comparing the health and community functioning of movers and non-movers at time 1 and 3 years later at time 2. SUBJECTS: The sample included 83 non-movers and 28 movers age 30 years and older who initially were living in nursing homes. The majority of residents had severe to profound mental retardation. The movers transferred to 15 community-based settings between 1 and 3 years (mean of 2 years) prior to the time 2 assessment. Assessments of residents at baseline and at follow-up included health measures (health status, health limits, mobility limits, days hospitalized and depression) and community functioning measures (adaptive functioning, maladaptive behaviour, community inclusion, day programme hours and monthly wages). RESULTS: Findings indicated that movers showed benefits in terms of improved health and community functioning. For movers, health status, mobility limitations, and community inclusion improved, while there were no significant changes for non-movers. This research corroborates previous research on the effects of deinstitutionalization and expands its implications to a group with severe disabilities. PMID- 10381241 TI - Mortality and morbidity among older adults with intellectual disability: health services considerations. AB - PURPOSE: Described is a study of the mortality and morbidity characteristics of 2752 adults with intellectual disability, age 40 and older, who died over a 10 year period in one American state. RESULTS: The main finding was that although individuals in the current generation of older adults with intellectual disability still generally die at an earlier age than do adults in the general population (average age at death: 66.1 years), many adults with intellectual disability live as long as their age peers in the general population. The results suggest that the longevity of adults with intellectual disability, whose aetiology is not attributable to organic causes, is progressively increasing. The results also confirm an increased longevity for adults with Down syndrome (average age at death: 55.8 years). Findings also showed that the causes of death for the study cohort were similar to those of the general older population, with cardiovascular, respiratory and neoplastic diseases among the most prominent causes of death. CONCLUSIONS: It was proposed that clinical and prophylactic health practices could have significant social and health care consequences for delaying the onset or minimizing the occurrence of life threatening diseases (and thus prolonging life) in adults with intellectual disability. It was suggested that clinical practices could be implemented that deter the onset and lessen the impact and burden of older age-related diseases and secondary conditions and that greater attention needs to be given to training of health care professionals in the area of geriatric medicine and intellectual disability. PMID- 10381242 TI - Measuring disability with parsimony. AB - PRIMARY OBJECTIVE: Health surveys, especially those for older persons, include numerous detailed items about disability. There has been little effort to develop a global disability item, that is, one question that covers the concept of disability briefly but well. This article discusses how parsimony can be achieved through a single item, or less desirably by reductions of detailed items. MAIN OUTCOME AND RESULTS: Results of three analyses on the issue of compact disability indicators, using public-use data sets (AHEAD, HRS, BRFSS), are presented. The analyses study relationships of global disability to both detailed disability items and global health. Overall, the results show that a global disability item has good coverage of specific disabilities and is distinct from self-rated health. CONCLUSIONS: Routine inclusion of a global disability item in surveys is recommended, and specific suggestions are made to aid its design. PMID- 10381243 TI - Alternative medicine, education and standards of publication and research. PMID- 10381244 TI - How safe is acupuncture? Developing the evidence on risk. PMID- 10381245 TI - The role of traditional healers in the fight against AIDS in Africa. PMID- 10381246 TI - Adverse events in acupuncture and moxibustion treatment: a six-year survey at a national clinic in Japan. AB - OBJECTIVES: Many of the frequently reported adverse effects of acupuncture are serious or severe such as pneumothorax, infection, spinal cord injury, or cardiac injury. However, reviewing published case reports does not provide enough information to assess the safety of acupuncture and moxibustion. In order to investigate adverse events of acupuncture, we reviewed all the relevant cases reported by the therapists at our clinic. SETTING: Over a 6 year period, in the national Tsukuba College of Technology Clinic in Japan, all the acupuncture and moxibustion therapists were required to report the cases of adverse events immediately upon recognition. RESULTS: A total of 84 therapists (13 preceptors and 71 interns) participated in the treatments. The total number of treatments was 65,482. Ninety four (0.14%) adverse events were reported. There were fourteen categories: failure to remove needles (27 cases), ecchymosis or hematoma without pain (9 cases), ecchymosis or hematoma accompanied by pain (8 cases), burn injury (7 cases), discomfort (7 cases), dizziness (6 cases), nausea or vomiting (6 cases), pain in the punctured region (6 cases), minor hemorrhage (4 cases), aggravation of complaint (4 cases), malaise (3 cases), suspected contact dermatitis (3 cases), fever (3 cases) and numbness in the upper extremities (1 case). CONCLUSION: No serious or severe cases such as pneumothorax, infection, or spinal cord injury were reported by the college preceptors and interns. The results indicate that serious or severe adverse events are rare in standard practice. We suggest that most severe or serious cases of adverse events caused by acupuncture reported in journals are actually cases of negligence. In the future, negligence should be discussed from the point of view of medical education and technical instruction for the therapists, and adverse reactions should be discussed from the point of view of incidence and prevention based on the result of further investigation. PMID- 10381247 TI - A controlled investigation of bodywork in multiple sclerosis. AB - OBJECTIVE: To determine whether a course of Feldenkrais bodywork would result in significant improvement in physical, mood symptoms and functioning in multiple sclerosis (MS) patients beyond the effects observed using a sham condition (nontherapeutic bodywork). DESIGN: The bodywork method used was the Feldenkrais method. Subjects were randomly assigned to 1 of 2 groups in a crossover design to control for order effects of treatment. Half of the subjects received 8 weeks of sham sessions followed by 8 weeks of Feldenkrais sessions. The other half of the subjects received Feldenkrais sessions first and then sham. All subjects completed the outcome measures prior to the first course of treatment, in between Feldenkrais and sham, and at study completion. SETTING: Participants were recruited from a regional MS clinic and were administered bodywork treatment and outcome measures in a bodywork practitioner's office. SUBJECTS: Twenty individuals with clinically definite MS and disability status scores between 2.0 and 6.0 participated. OUTCOME MEASURES: Nine-hole pegboard test of hand dexterity, Hospital Anxiety and Depression Scale, MS self-efficacy scale, MS Symptom Inventory, MS Performance Scales, and the Perceived Stress Scale. RESULTS: The only significant differences were observed for perceived stress and lowered anxiety after Feldenkrais sessions. There were nonsignificant trends toward higher self-efficacy after both Feldenkrais and sham sessions. MS symptoms, levels of functional ability, and upper extremity performance were not affected by Feldenkrais or sham sessions. PMID- 10381248 TI - Enhanced thermogenesis in rats by a composite Indian herbal preparation-I and its mechanism of action. AB - OBJECTIVE: A composite Indian herbal preparation-I (CIHP-I) containing ingredients derived from 7 different plants and asphalt was tested for its adaptogenic activity and its mechanism of action was investigated. DESIGN: CIHP-I was tested using the cold-hypoxia-restraint (C-H-R) animal model in which the restrained rats were exposed to 5 degrees C at 428 mm Hg atmospheric pressure. Rectal temperature (Trec) of the rats was continuously monitored during the exposure and the recovery periods. The time for fall of Trec to 23 degrees C and its recovery to 37 degrees C were used as indices of endurance and the adaptogenic activity. Carbohydrate and lipid parameters were investigated to find out the nature of fuel being used during thermogenesis. RESULTS: After 12 weeks of administration of an oral dose of 7.5 mg/kg(-1)/day(-1), CIHP-I was found to possess significant adaptogenic activity. CIHP-I helped improve resistance to C-H R induced hypothermia (Trec 23 degrees C) in animals by increased mobilization of free fatty acids (FFA) from adipose tissue. Blood glucose and muscle glycogen levels were maintained. CIHP-I treatment restricted the release of creatine phosphokinase (CPK) into the circulation during C-H-R exposure. CONCLUSIONS: The results suggested that CIHP-I is a strong adaptogen. It improved cold resistance during C-H-R exposure and enhanced recovery from hypothermia. The energy dependent cell membrane permeability was maintained. Stored lipids were mobilised and possibly used for thermogenesis in preference to carbohydrates. PMID- 10381249 TI - Short-term outcomes of treatment for musculoskeletal disorders in a hospital based alternative and complementary medicine clinic. AB - OBJECTIVE: Musculoskeletal (MS) disorders are among the more common conditions for which patients seek relief. The treatment of choice for MS disorders in Western medicine is the administration of nonsteroidal anti-inflammatory (NSAID) agents. However, patients in increasing numbers are seeking out complementary/alternative medicine (CAM) for MS disorders. This report presents the results of a systematically conducted, short-term evaluation of MS patients seen in a hospital-based outpatient clinic that provides acupuncture and other alternative modalities. DESIGN: During a 1.5-year period, 797 patients in our CAM clinic completed a structured questionnaire on their initial visit. The questionnaire elicited demographic and health history information and information regarding the nature and severity of the patients' health complaints. One-month after this baseline measurement and the beginning of CAM treatment, patients rated their complaint severity, improvement in the condition being treated and helpfulness of the CAM treatments. Of the 797 patients, 398 were treated for MS complaints and provide the basis for this report. RESULTS: At the 1-month follow up, severity of primary complaints diminished an average of 2.0 points (SD = 2.4) on a 10-point numerical scale (P<0.001). Thirty percent of the patients reported severity reduction to 50% or more of baseline. Although self-reported improvement and change in severity were moderately correlated, baseline severity was inversely related to therapeutic helpfulness (P = 0.013). Improvement increased with number of treatments (P<0.001); however, due to time constraints, treatment frequency was limited. CONCLUSIONS: Nearly half of the patients visiting our clinic sought relief for MS complaints. Our findings suggest that acupuncture treatment provides significant improvement of presenting symptoms within a short time period. Although discerning the long-term treatment effects was not the intent of this study, these findings contribute toward a consensus opinion that CAM therapies can help alleviate the pain and discomfort of MS disorders. PMID- 10381250 TI - Teaching an integrated approach to complementary, alternative, and mainstream therapies for children: a curriculum evaluation. AB - BACKGROUND: Increasing numbers of patients seek information about complementary and alternative medicine (CAM) from their primary physicians. We sought to evaluate our 4-year old curriculum integrating mainstream and CAM care for common outpatient pediatric problems within a family medicine residency. DESIGN: Cross sectional survey. METHODS: Subjects included current (1998) third-year residents and recent graduates from our program and nearby University of Washington affiliated family medicine residency programs. The survey included items on training experiences, knowledge, attitudes and behavior regarding CAM. RESULTS: Among the 18 respondents from our program and 21 from comparison programs, the average age was 32 years and one-third were male. Over 80% of respondents felt that residencies should provide training in CAM. Substantial numbers of respondents from all programs recommended CAM therapies to patients in the past year. All respondents had recommended special diets and nutritional supplements; more than 50% recommended herbal remedies, acupuncture, meditation or progressive relaxation, massage or home remedies. Respondents from all groups had similar attitudes and knowledge about integrative medicine; those from the intervention program were more likely than comparison respondents to agree that their residency training had prepared them to answer patients' questions about CAM (50% vs. 19%, p = 0.04). CONCLUSIONS: Primary care residents increasingly seek training to answer patients' questions and are already recommending a variety of CAM therapies. Primary care residencies need to develop and evaluate responsible, evidence-based curricula integrating mainstream and CAM therapies. PMID- 10381251 TI - Emerging educational needs of an emerging discipline. PMID- 10381253 TI - Attitudes of medical and nonmedical students toward orthodox and complementary therapies: is scientific evidence taken into account? AB - Medical and nonmedical students completed a questionnaire indicating how willing they would be to try various therapies for treatment. Both groups assessed more traditional complementary practices such as homeopathy and acupuncture as similar to more orthodox treatments such as physiotherapy or prescribed diets. Both groups appeared not to differentiate between established techniques (physiotherapy) and less tested techniques (yoga). Furthermore, neither group seemed particularly concerned about the scientific evaluations of treatments. PMID- 10381252 TI - Vanadium: a review of its potential role in the fight against diabetes. AB - The potential role of vanadium in human health is described as a building material of bones and teeth. However, another very interesting and promising application for vanadium in human health emerges from recent studies that evaluated the role of vanadium in the management of diabetes. Vanadium is present in a variety of foods that we commonly eat. Skim milk, lobster, vegetable oils, many vegetables, grains and cereals are rich source of vanadium (>1 ppm). Fruits, meats, fish, butter, cheese, and beverages are relatively poor sources of vanadium. The daily dietary intake in humans has been estimated to vary from 10 microg to 2 mg of elemental vanadium, depending on the environmental sources of this mineral in the air, water, and food of the particular region tested. In animals, vanadium has been shown essential (1-10 microg vanadium per gram of diet). There is only circumstantial evidence that vanadium is essential for humans. However, in doses ranging from 0.083 mmol/d to 0.42 mmol/d, vanadium has shown therapeutic potential in clinical studies with patients of both insulin dependent diabetes mellitus (IDDM) and noninsulin-dependent diabetes mellitus (NIDDM) type. Although vanadium has a significant biological potential, it has a poor therapeutic index, and attempts have been made to reduce the dose of vanadium required for therapeutic effectiveness. Organic forms of vanadium, as opposed to the inorganic sulfate salt of vanadium, are recognized as safer, more absorbable, and able to deliver a therapeutic effect up to 50% greater than the inorganic forms. The goal is to provide vanadium with better gastrointestinal absorption, and in a form that is best able to produce the desired biological effects. As a result, numerous organic complexes of vanadium have been developed including bis(maltolato)oxovanadium (BMOV), bis(cysteinamide N-octyl)oxovanadium known as Naglivan, bis(pyrrolidine-N-carbodithioato)oxovanadium, vanadyl-cysteine methyl ester, and bis-glycinato oxovanadium (BGOV). The health benefits of vanadium and the safety and efficacy of the available vanadium supplements are discussed in this review. PMID- 10381254 TI - The practice of complementary medicine outside the National Health Service. PMID- 10381255 TI - Cardiovascular responses to a hot tub bath. AB - This study was conducted to determine the cardiovascular effects of 15 minutes of hot tub immersion at 39 degrees C. Five college-age subjects (4 males and 1 female) volunteered to participate in this study. Assessments were made while sitting first in a chair for 5 minutes and then in the hot tub for 15 minutes. Oxygen consumption (VO2) and cardiac output (Q) measurements were made using a Medical Graphics CPX/D metabolic analyzer. Cardiac output was determined at minute 15 using the indirect CO2 rebreathing procedure. The data were analyzed using the analysis of variance with repeated measures, which indicated that at minute 15, heart rate (HR) and Q were increased, which increased VO2. The increase in Q was due to the heart rate (HR) response and the decrease in systemic vascular resistance (SVR). Mean arterial pressure (MAP) and systolic blood pressure (SBP) were decreased while double product (DP) was increased. There were no changes in stroke volume (SV) or arteriovenous oxygen difference (a vO2 diff). These findings indicate that the HR and Q responses are necessary to the increase in metabolism (VO2). Hot tube use within these time and temperature constraints should reduce concern over hot tub safety in college-age subjects. PMID- 10381256 TI - Cutaneous rat wounds express c49a, a novel gene with homology to the human melanoma differentiation associated gene, mda-7. AB - We have used DD-PCR (differential display-polymerase chain reaction) to identify new genes that are over- or underexpressed during wound repair. DD-PCR performed on excisional wounds identified the expression of rat c49a. Cloning and sequence analysis of the rat c49a gene revealed high homology to a novel human melanoma differentiation associated gene, mda-7. The human mda-7gene isolated from melanoma cell lines, has been linked with human melanoma differentiation, and growth suppression. Moreover, transfection of human mda-7 constructs into human tumor cells suppresses the growth and colony formation of tumor cells from diverse origins. To confirm and relatively quantitate expression of rat c49a gene during repair, specific primer, reduced cycle RT-PCR (reverse transcription-PCR) was performed. RT-PCR showed an approximately 9 to 12-fold elevation of rat c49a mRNA at 12 h to 5 days above nonwounded controls that gradually decreased to approximately 1.5 to 3-fold by day 14. Cloning and sequence analysis of the entire 1200 base pair c49a gene product showed 78% nucleotide homology to human mda-7. Immunohistochemistry studies localized rat C49A expression primarily to fibroblast-like cells at the wound edge and base. The marked up-regulation of rat c49a transcripts during the inflammatory and early granulation tissue phases of wound repair where cellular processes such as re-epithelialization, angiogenesis, and fibroplasia predominate--suggest that c49a is associated with proliferation of fibroblasts in wound healing. PMID- 10381257 TI - Metaxin 1 interacts with metaxin 2, a novel related protein associated with the mammalian mitochondrial outer membrane. AB - A recently described protein, metaxin 1, serves as a component of a preprotein import complex in the outer membrane of the mammalian mitochondrion. A yeast two hybrid screen with metaxin 1 as bait has now identified a novel protein, which we have termed metaxin 2, as a metaxin 1-binding protein. Metaxin 2 shares 29% identity with metaxin 1 at the amino acid level, but metaxin 2, unlike metaxin 1, lacks a C-terminal mitochondrial outer membrane signal-anchor domain. Two C. elegans hypothetical proteins, CelZC97.1 and CelF39B2.i, share high sequence similarity with metaxin 2 and metaxin 1, respectively, and likely represent the C. elegans orthologs. Affinity-purified antibodies against metaxin 2 were prepared against the recombinant protein produced in E. coli and were used to analyze the subcellular distribution of metaxin 2. In subcellular fractions of mouse liver, a 29 kD immunoreactive protein, consistent in size with the predicted translation product of metaxin 2 cDNA, was found solely in mitochondria. Alkali extraction of mitochondria indicated that metaxin 2 is peripherally associated with mitochondrial membranes. Metaxin 2 in intact mitochondria was susceptible to digestion with proteinase K, indicating that metaxin 2 is located on the cytosolic face of the mitochondrial outer membrane. Finally, baculoviruses encoding a His6-tagged metaxin 2 and an untagged metaxin 1 lacking its C-terminal transmembrane domain were produced and used separately or in combination to infect Sf21 insect cells. Metaxin 1 bound to a Ni2+-chelate affinity column only in the presence of metaxin 2, indicating that metaxin 1 and metaxin 2 interact when overexpressed in insect cells. These results suggest that metaxin 2 is bound to the cytosolic face of the mitochondrial outer membrane by means of its interaction with membrane-bound metaxin 1, and that this complex may play a role in protein import into mammalian mitochondria. PMID- 10381258 TI - Overexpression of ErbB2 impairs ligand-dependent downregulation of epidermal growth factor receptors via a post-transcriptional mechanism. AB - The mechanism by which ErbB2 exerts its oncogenic effect is poorly defined. In this article we show that ErbB2 co-expression slows ligand-dependent growth factor receptor downregulation in NIH 3T3 transfectants. Ligand dependence of cell growth and MAP kinase signaling are retained in epidermal growth factor receptor (EGFR) transfectants but are abolished in ErbB2-expressing cells, which grow and signal constitutively. In association with this phenomenon, we have noticed that ErbB2-expressing cells contain increased amounts of EGFR, which is hyperphosphorylated. EGFR overexpressors do not contain increased levels of ErbB2, however, tending to exclude a transfection artifact caused by saturation of receptor processing. EGF treatment of EGFR transfectants results in more rapid EGFR downregulation than occurs in ErbB2 transfectants, but Northern blot analysis demonstrates reduced basal EGFR gene expression in ErbB2 transfectants. We conclude that ErbB2 expression impairs EGFR downregulation via a posttranscriptional mechanism and propose that ErbB2 overexpression may sensitize tumor cells to the mitogenic effects of heterologous growth factors by retarding degradation of liganded heterodimers. PMID- 10381260 TI - Functional analysis of DNA sequences located within a cluster of DNase I hypersensitive sites colocalizing with a MAR element at the upstream border of the chicken alpha-globin gene domain. AB - We have cloned and sequenced a genomic DNA fragment of chicken containing a cluster of DNase I hypersensitive sites (DHS) located 11-15 kb upstream from the first gene of the alpha-globin gene domain and including a constitutive DHS flanked by two erythroid-specific ones. A 1.2-kb subfragment of the DNA fragment under study located upstream to the constitutive DHS and colocalizing roughly with one of the erythroid-specific DHS was shown to possess the properties of a matrix association region (MAR). The cloned DNA sequences were tested for their ability to serve as promoters and/or influence transcription from the promoter of the alphaD globin gene. In the region studied, we did not find any promoters or enhancers that were active in erythroid cells. The whole DNase I hypersensitive region and some of its subfragments showed a silencing effect when placed downstream from the reporter gene. The expression of the reporter gene was completely abolished, however, when these DNA fragments were placed between the alphaD promoter and the reporter gene. Thus, they seem to act as transcription "terminators." Numerous polyadenylation signals (AATAAA) and an AT-rich palindrome were found within the sequenced DNA fragment. These observations are discussed within the frame of the hypothesis postulating that continuous transcription is essential for maintaining the active status of genomic domains. Furthermore, it is suggested that the DNA fragment studied contains a negative control element that keeps globin genes silent within the chromatin domain permanently open in nonerythroid cells. PMID- 10381259 TI - Antisense inhibitory effect: a comparison between 3'-partial and full phosphorothioate antisense oligonucleotides. AB - Phosphorothioate (PS) antisense oligonucleotides are currently used to inhibit many cell functions both in vivo and in vitro. However, these modified oligos provide reasonable sequence specificity only within a narrow concentration range. To overcome such a limitation we synthesized antisense oligomers, partially phosphorothioated, targeted against the human N-myc mRNA. We utilized such modified oligomers in a human neuroblastoma cell line where the N-myc gene expression was very high, and compared them to full phosphorothioate oligonucleotides. Both full PS and partial PS antisense oligos produced a maximum reduction in target mRNA after 6 h of treatment. They were able to maintain a good level of inhibition for 20 h only at high concentration. While partial PS oligos produced a dose dependent and sequence specific inhibition of N-myc mRNA, full PS molecules suffer from some disadvantages at the highest concentration used. Our results showed that partial PS molecules were capable of reducing gene expression showing a greater sequence specificity over a far broader concentration range. For this reason we conclude that partial PS antisense oligos, with respect to full PS antisense oligos, might be particularly useful for studying gene function. PMID- 10381261 TI - Phospholipase C-beta and ovarian sex steroids in pig granulosa cells. AB - We compared the membrane effects of estradiol, progesterone, and androstenedione in a single experimental model, the ovarian granulosa cells collected from immature Large White sows. We measured changes in cytosolic free calcium concentration ([Ca2+]i) in confluent Fura-2 loaded cells. We used pharmacological tools and polyclonal phospholipase C-beta (PLC-beta) antibodies. Each steroid (0.1 pM to 1 nM) transiently increased intracellular calcium concentration ([Ca2+]i) within 5 sec. They mobilized Ca2+ from the endoplasmic reticulum as shown by using two phospholipase C inhibitors, neomycin and U-73122. Ca2+ mobilization involved PLC-beta1 for progesterone, PLC-beta2 for estradiol and PLC beta4 for androstenedione. A pertussis toxin-insensitive G protein was involved in the effects of progesterone on Ca2+ mobilization whereas estradiol and androstenedione effects were mediated via a pertussis toxin-sensitive G-protein. Ca2+ influx from the extracellular milieu was involved in the increase in [Ca2+]i induced by progesterone and estradiol, but not by androstenedione. Influx of Ca2+ was independent of Ca2+ mobilization from calcium stores, and it was suggested that L-type Ca2+ channels for estradiol and T-type Ca2+ channels for progesterone were involved. The three steroids had no effect on cAMP. Rapid effects of progesterone, estradiol, and androstenedione involved a direct action on cell membrane elements such as PLC-beta, G-proteins, and calcium channels, and these mechanisms were hormone-specific. PMID- 10381262 TI - Urokinase expression and binding activity associated with the transforming growth factor beta1-induced migratory and invasive phenotype of mouse epidermal keratinocytes. AB - Transforming growth factor beta1(TGF-beta1) is a stimulator of malignant progression in mouse skin carcinogenesis. TGF-beta1 exerts a differential effect on cultured nontumorigenic (MCA3D cell line) and transformed (PDV cell line) keratinocytes. Whereas MCA3D cells are growth arrested and committed to die in the presence of the factor, it induces a reversible epithelial-fibroblastic conversion in PDV cells. This conversion is associated in vivo with a squamous spindle cell carcinoma transition. Here we have investigated the role of urokinase (uPA) during malignant progression of transformed epidermal keratinocytes. We show that the levels of uPA expression/secretion, and the uPA binding activity to the cell surface, correlate with the invasive and malignant potentials of mouse epidermal cell lines. TGF-beta1 enhanced uPA production, the number of uPA cell surface binding sites, and the expression of the plasminogen activator inhibitor PAI-1, in transformed PDV cells, but had no major effect on nontumorigenic MCA3D keratinocytes. Increased uPA production depended on the presence of the factor in the culture medium and occurred concomitantly to the stimulation of the migratory and invasive abilities of PDV cells. Synthetic peptides containing the amino terminal sequence of the mature mouse uPA inhibited the binding of uPA to the cell surface and decreased TGF-beta1-induced cell motility and invasiveness. These results demonstrate that the uPA system mediates at least part of the migratory and invasive phenotype induced by TGF-beta1 in transformed keratinocytes, and suggest a role for uPA on the changes that lead to the appearance of spindle carcinomas. PMID- 10381263 TI - Inhibition of testosterone secretion by digitoxin in rat testicular interstitial cells. AB - Both in vivo and in vitro experiments were conducted to determined the effects of digitoxin on the secretion of testosterone, and its underlying mechanisms including testicular adenosine 3':5'-cyclic monophosphate (cAMP), and the activities of steroidogenic enzymes. Male rats were injected with digitoxin, human chorionic gonadotropin (hCG), or hCG plus digitoxin via a jugular catheter. Blood samples were collected immediately before and at 30 and 60 min after the challenge, and analyzed for testosterone by radioimmunoassay. In an in vitro study, rat testicular interstitial cells were isolated and incubated with digitoxin, hCG, 8-bromo-cAMP (8-Br-cAMP), digitoxin plus hCG, or digitoxin plus 8 Br-cAMP at 34 degrees C for 1 h. The media were collected and analyzed for testosterone. For studying cAMP accumulation, testicular interstitial cells were incubated for 1 h in the medium containing isobutyl-1-methylxanthine (IBMX) and different doses of digitoxin with the absence or presence of hCG. After incubation, cells were processed for determining cAMP content. Intravenous injection of digitoxin decreased hCG-stimulated, but not basal, plasma testosterone levels. Administration of digitoxin in vitro resulted in an inhibition of both basal and hCG- as well as 8-Br-cAMP-stimulated release of testosterone. In addition, digitoxin diminished hCG-stimulated cAMP accumulation in rat testicular interstitial cells. Furthermore, digitoxin inhibited the activity of cytochrome P450 side chain cleavage enzyme (P450scc) but failed to affect the activities of other steroidogenic enzymes. Taken together, these results suggest that the acute inhibitory effect of digitoxin on the testosterone production in testicular interstitial cells involves, at least partly, an inefficiency of post-cAMP events, and a decrease of P450scc activity. PMID- 10381264 TI - Involvement of intracellular signaling factors in the serum-enhanced Ca2+-binding protein regucalcin mRNA expression in the cloned rat hepatoma cells (H4-II-E). AB - The involvement of signaling factors, which are related to serum component, on the regucalcin mRNA expression in the cloned rat hepatoma cells (H4-II-E) was investigated. The change in regucalcin mRNA levels was analyzed by Northern blotting using rat liver regucalcin complementary DNA (0.9 kb of open reading frame). H4-II-E cells were cultured for 2 or 6 h in a medium containing various reagents in the presence of serum (10% fetal bovine serum) after the subconfluent with 3-day-culture. The regucalcin mRNA expression was significantly increased by serum addition. This increase was clearly inhibited by the presence of EGTA (10( 3) M), A23187 (10(-6) M), trifluoperazine (10(-5) M), staurosporine (10(-7) M), or genistein (10(-5) M) with 6-h-culture, although the beta-actin mRNA expression was not altered by the reagents. Meanwhile, the regucalcin mRNA expression was significantly stimulated by the addition of Bay K 8644 (2.5 x 10(-6) M) in the presence of serum. This effect was also seen in the presence of genistein (10(-5) M). The present study suggests that the regucalcin mRNA expression is mediated through signaling pathways which are partly involved in Ca2+-dependent protein kinases and tyrosine kinase in H4-II-E hepatoma cells. PMID- 10381265 TI - Dystonia musculorum mutation and myosin heavy chain expression in skeletal and cardiac muscles. AB - This study evaluated the influence of dystonia musculorum (dt) mutation, characterized by spinocerebellar fibers degeneration, on cardiac and skeletal muscles: one respiratory (diaphragm, Dia), three masticatory (anterior temporalis, AT; masseter superficialis, MS; and anterior digastric, AD), one hindlimb (soleus, S), tongue (T), and one cardiac (ventricle, V). Body and muscle weight, muscle protein content, and myosin heavy chain (MHC) isoforms relative expression were then compared in dt mutant mice and in normal mice, according to sex. Male body and muscle weight was always greater than that of females, but there was no specific muscle difference in females. dt mutant mice showed a reduced whole body growth but no specific muscle atrophy, as well as a global decrease in muscle protein content that made muscles more fragile. dt mutation induced a global reduction of muscle protein concentration, whereas a general influence of sex could not be disclosed. Concerning MHC relative composition, all the muscles were fast-twitch: Dia, AT, MS, AD, S, and T expressed predominantly the fast type 2 MHC isoforms, whereas V contained only MHC alpha, also a fast MHC. Female muscles were slower than male muscles, except for S, which was faster. However, classification of muscles in terms of shortening velocity was very different in normal males and females. In other respects, dt mutant muscles were slower and consequently more fatigue resistant than normal, except for S, which became faster and less fatigue resistant. dt mutation exhibits then a specific effect on this continually active postural muscle. In the other muscles, global increased fatigue resistance could constitute an adaptive response to work requirements modifications linked to the muscle damage. It should be noted that a developmental MHC (neonatal) was present in female dt AD. Innervation, which influences muscle structure, is altered in dt mutant and could be another causal factor of the fast-to-slow MHC switches. It appears that dystonin, the dt gene product, is very important in maintaining the structural integrity of both cardiac and skeletal muscle and in its absence, the muscle becomes more fragile and is damaged by modified activity. PMID- 10381266 TI - Biochemical and morphological changes in the nuclear matrix prepared from apoptotic HL-60 cells: effect of different stabilizing procedures. AB - Apoptotic cell death is characterized by deep morphological changes that take place in the nucleus. It is unclear whether modifications also occur in the nuclear matrix, a mainly proteinaceous structure that conceivably acts as a nuclear framework. We have investigated whether biochemical and morphological alterations of the nuclear matrix prepared from apoptotic HL-60 cells were dependent on the manipulations to which isolated nuclei were subjected before DNase I digestion and 2 M NaCl extraction. Our results showed that the stabilizing procedures employed to preserve the inner fibrogranular network and nucleolar remnants of the matrix (i.e., a 37 degrees C incubation; exposure to sodium tetrathionate at 4 degrees C; exposure to sodium tetrathionate at 37 degrees C) had no effect on the protein recovery of apoptotic nuclear matrices, which was always approximately two- to fivefold less than in control matrices. Moreover, one- and two-dimensional gel analysis of nuclear matrix proteins showed that, in apoptotic samples, striking quantitative changes were present, as compared with controls. Once again, these changes were seen irrespective of the stabilizing procedures employed. Also, transmission electron microscope analysis showed similar morphological alterations in all types of apoptotic nuclear matrices. By contrast, the immunofluorescent distribution of the 240-kDa NuMA protein seen in apoptotic samples was more sensitive to the stabilizing treatments. Our results indicate that the biochemical and morphological changes of the apoptotic nuclear matrix are largely independent of the isolation protocols and strengthen the contention that destruction of the nuclear matrix network is one of the key events leading to apoptotic nuclear destruction. PMID- 10381267 TI - Regulation of testosterone secretion by prolactin in male rats. AB - The goal of this study was to characterize the mechanism by which hyperprolactinemia alters testosterone production in rat testicular interstitial cells (TICs). Hyperprolactinemia was induced by grafting 2 anterior pituitary (AP) glands under the subcapsular space of the kidney in experimental rats. Control rats were grafted with brain cortex (CX). Six weeks post-grafting, rats were challenged with human chorionic gonadotropin (hCG) then, the changes in either plasma testosterone or luteinizing hormone was measured. Additionally, TICs were isolated and challenged in vitro with hCG or prolactin, and the testosterone release measured by radioimmunoassay. Further investigation in signal transduction as intracellular 3':5' cyclic adenosine monophosphate (cAMP) production was observed under a regulation of forskolin or SQ22536. After the challenge of hCG or GnRH, the AP-grafted rats showed a suppressed response in testosterone release as compared to those in the CX-grafted group. The in vitro data from the AP-grafted rats compared to the CX-grafted animals showed a diminished response in testosterone release upon hCG stimulation. Administration of forskolin or SQ22536 disclosed dysfunction of adenylate cyclase in TICs from the AP-grafted rats. When 8-Br-cAMP was incubated with TICs, the testosterone production was lower in the AP-grafted compared to the CX-grafted group. These results suggest that in addition to adenylate cyclase dysfunction, inefficiency of post-cAMP pathways are also involved in the hypogonadism elicited by hyperprolactinemia in rats. PMID- 10381268 TI - Expression of SET is modulated as a function of cell proliferation. AB - We explored a biological role of SET as it relates to cell proliferation and differentiation. Immunohistochemical staining demonstrated that the expression of SET was ubiquitous and diffuse over the whole embryo on gestational day 15. At a later stage of development, SET was expressed at relatively lower levels and localized to specific tissues and cells. On embryonic day 19, specific SET immunoreactivity was found in the epithelium of skin, respiratory tract, intestine, and retina as well as in muscle and cartilage. In these cells SET was stained mostly in the nucleus, which was supported indirectly by nuclear transport of enhanced green fluorescence protein-SET fusion proteins in ECV304 endothelial cells. Set mRNA expression was further confirmed in various cultured cells, including NIH 3T3 cells, L6 myoblast cells, human umbilical vein endothelial cells, and ECV304 cells. Using F9 teratocarcinoma cell lines, which were stimulated to differentiate into the two different cell lineages of parietal and visceral endoderm, we have further examined the role of SET. The expression of set mRNA and SET protein was diminished about three-fold in both differentiated endoderm cells compared to the undifferentiated F9 cells. However, when F9 cells were subjected to serum starvation, reduction of set mRNA abundance also took place at a similar level to that observed in response to differentiation. Consistent with this, quiescent L6 myoblast showed a marked downregulation of set mRNA compared to proliferating cells. These results suggest that SET is involved mainly in the regulation of cell proliferation rather than differentiation during embryonic development. PMID- 10381269 TI - Differential targeting of protein kinase CK2 to the nuclear matrix upon transient overexpression of its subunits. AB - Modest dysregulation of CK2 has been shown to enhance the oncogenic potential in transgenic models of cancer. Since nuclear matrix serves as an anchor for CK2 and plays a key role in growth-related activities, we examined the effects of CK2 overexpression on its signaling to the nuclear matrix. Expression plasmids pCI CK2alpha, pCI-CK2beta, and the bicistronic pCI-CK2alphabeta containing full length cDNAs encoding the various subunits were employed to transiently transfect two cell lines, BPH-1 and COS-1. Cytosol from transfected BPH-1 cells containing alpha or beta or alpha + beta or alphabeta showed a modest increase in CK2 activity by 26%, 1%, 20%, and 17%, respectively, over that in the controls transfected with pCI vector. However, the corresponding increase in CK2 activity in the NM fraction was 156%, 8%, 147%, and 152%, respectively. Immunoblot analysis of the CK2 in the NM accorded with these data. Similar results were obtained with COS-1 cells or other expression vectors. The results suggest that moderate overexpression of CK2 in the cells evokes a differential several-fold enhancement in NM associated CK2 relative to that in the cytosol. This process may have a bearing on the functional signaling of this kinase in relation to its possible role in oncogenesis. PMID- 10381270 TI - Role of hypoxia and extracellular matrix-integrin binding in the modulation of angiogenic growth factors secretion by retinal pigmented epithelial cells. AB - The retinal pigmented epithelium (RPE) is a monolayer of polarized cells located between retinal photoreceptors and blood vessels of the choroid. The basal surface of RPE cells rests on Bruch's membrane, a complex extracellular matrix structure which becomes abnormal in several disease processes, including age related macular degeneration (AMD). Ruptures or abnormalities in Bruch's membrane are frequently accompanied by choroidal neovascularization. Disturbed interaction of RPE cells with their extracellular matrix (ECM) could play a role in this process. The present study was undertaken to examine the complex interactions between hypoxia, integrin, and ECM in the regulation of RPE functions. Antibody blocking experiments demonstrated that RPE cell adhesion to vitronectin is mediated primarily through alphavbeta5 and adhesion to fibronectin occurs through alpha5beta1. RPE adhesion to immobilized laminin demonstrated highest level of non-RGD-mediated adhesion as compared to that with collagen IV or the RGD matrices such as vitronectin (alphavalpha5) , fibronectin (alpha5beta1), or thrombospondin (alpha5beta1 + alphavbeta5). Addition of soluble vitronectin, or fibrinogen to RPE cell cultures resulted in a small to moderate increase in VEGF and FGF2 in the media, while each of these growth factors was dramatically increased after addition of thrombospondin 1 (TSP1). In contrast, soluble fibronectin resulted in differential upregulation of VEGF but not FGF2. Similarly, immobilized TSP1 resulted in differential greater upregulation in VEGF but not FGF2 release from RPE as compared to other ECMs under either normoxic or hypoxic conditions. Additionally, hypoxia resulted in a time-dependent increase in VEGF, but not FGF2 release in the media. RPE cells grown on TSP1-coated plates showed increased VEGF and FGF2 in their media compared to cells grown on plates coated with type IV collagen, laminin, vitronectin, or fibronectin. The TSP1 induced increase in secretion of growth factors was partially blocked by anti alpha5beta1, anti-alphavbeta3, and anti-alphavbeta5 antibodies indicating that it may be mediated in part by TSP1 binding to those integrins. These data suggest that alterations in oxygen levels (hypoxia/ischemia) and ECM of RPE cells, a prominent feature of AMD, can cause increased secretion of angiogenic growth factors that might contribute to the development of choroidal neovascularization. These data also suggest the potential modulatory role of VEGF release from RPE by ECM and alphavbeta5 and alpha5beta1 integrins. PMID- 10381271 TI - An historical perspective on atherosclerosis research in Germany. PMID- 10381272 TI - Concordant upregulation of type II-TGF-beta-receptor, the cyclin-dependent kinases inhibitor P27Kip1 and cyclin E in human atherosclerotic tissue: implications for lesion cellularity. AB - Atherosclerosis is a 'response-to-injury' process associated with chronic inflammation, tissue repair and a considerable cell turnover. These growth related processes are controlled by the 'cell cycle clock' which is composed of cyclin-dependent kinases (Cdks), their activating subunits, the cyclins, and by inhibitors of Cdks (Ckis). P27 is a Cki which associates with cyclin A-Cdk2, cyclin D-Cdk4 and with cyclin E (CE)-Cdk2 complexes thereby abrogating their catalytic activity leading to potent inhibition of late G1 to S-phase transition. Furthermore, TGF-beta1 mRNA and immunoreactivity are locally increased in atherosclerotic lesions. Since TGF-beta1 growth suppressive function in the late G1 phase may be mediated by p27, blocking the catalytic activity of CE-Cdk2 complexes, via the stimulation of TGF-beta-RI and TGF-beta-RII, we investigated the topographical association between TGF-beta-RI, TGF-beta-RII, P27Kip1 and CE by immunohistochemistry in coronary artery segments without atherosclerosis and carotid atheromatous plaques of 11 patients undergoing carotid endarterectomy. P27-immunoreactivity was present in 11/11 atherosclerotic (92.7 +/- 3.3% of the cells) and 5/5 control (80.9 +/- 3.7% of the cells; P < 0.002 versus control) specimens and localized to nuclei of macrophages (CD68-positive), vascular smooth muscle cells (alpha-actin positive), T-lymphocytes (CD3-positive) as well as to the nuclei of endothelial cells. In the atherosclerotic tissue, TGF-beta-RI and TGF-beta-RII-immunoreactivity was present in 11/11 specimens and localized to inflammatory cells and to cells with VSMC-like-morphology. TGF-beta-RI immunoreactivity was present in 87.4 +/- 5.3% (controls 75.3 +/- 7.48%; n.s.) and TGF-beta-RII-immunoreactivity was present in 83.7 +/- 6.8% (controls 39.5 +/- 7.3%; P < 0.002) of the cells. Double immunolabeling, and investigation of serial sections revealed co-expression of TGF-beta-RI and TGF-beta-RII in virtually all cells positive for P27. In the atherosclerotic specimens, CE-immunoreactivity was present in all specimens in macrophages (CD68-positive), vascular smooth muscle cells (alpha-actin positive) and in endothelial cells in 12.58 +/- 13.58% of the nuclei whereas in the controls CE staining was restricted to 0.19 +/- 0.43% of the cells (P < 0.001). Importantly, as shown by immunofluorescent double labeling, we found cells expressing P27 that were simultaneously positive for CE. In summary, the present study provides evidence that TGF-beta1 present in human atherosclerotic tissue may mediate its growth suppressive activity also by p27, blocking the activity of CE-Cdk2 complexes. Quantitative analysis revealed that TGF-beta-RII, p27 and CE are concordantly upregulated in the atherosclerotic tissue with chronic inflammation, supporting the view that TGF-beta1, p27 and CE may play an important role in the processes associated with chronic inflammation and cell turnover in advanced human atherosclerotic plaques. Taken together, these results provide a possible link between the chronic inflammation associated with advanced atherosclerosis, the effects of extracellular growth factors and cell cycle control. PMID- 10381273 TI - Expression of monocyte chemoattractant protein-1 cDNA in vascular smooth muscle cells: induction of the synthetic phenotype: a possible clue to SMC differentiation in the process of atherogenesis. AB - In the arterial wall, smooth muscle cells (SMC) normally exist in a quiescent, differentiated state, representing the contractile phenotype. During the development of atherosclerosis SMC change towards the synthetic phenotype going along with proliferation, chemotactic response and increased monocyte binding. Expression of monocyte chemoattractant protein-1 (MCP-1), a potent chemoattractant for monocytes, has been shown to be among the earliest events in atherogenesis. We investigated the effect of MCP-1 on differentiated and dedifferentiated SMC. Differentiation of SMC was induced using Matrigel as a matrix for cultivation. MCP-1 was expressed in SMC by means of a recombinant adenovirus. Expression of MCP-1 led to dedifferentiation of SMC as demonstrated by induction of cytokeratin 18, a marker for the synthetic phenotype. Concurrently, migration was only detectable in MCP-1 expressing cells, whereas SMC infected with a control virus, coding for the nuclear-targeted lacZ gene showed no migration. The expression of intercellular adhesion molecule-1 (ICAM-1) could be demonstrated in synthetic SMC and was induced after infection of differentiated cells with recombinant adenovirus, coding for MCP-1 (AdMCP-1). Expression of ICAM-1 was associated with a tenfold higher monocyte binding compared to lacZ infected cells. Our data suggest that MCP-1 plays an important role for SMC in the functional switch from the contractile to the synthetic phenotype in the course of atherogenesis. PMID- 10381274 TI - Colony stimulating factors modulate the transcription of type VIII collagen in vascular smooth muscle cells. AB - Colony stimulating factors belong to a family of cytokines that regulate proliferation in macrophages and other vascular cell types. They have been implicated in the inflammatory-fibroproliferative response of atherosclerosis. The present study was undertaken to assess the effect of granulocyte-macrophage and macrophage colony stimulating factors on the transcription of type VIII collagen by vascular smooth muscle cells and their potential relevance for the expression of collagen in atherosclerotic lesions. The influence of colony stimulating factors was studied in relation to transforming growth factor beta1, the factor exhibiting the most potent effect on collagen metabolism. Northern blot experiments showed that treatment with both colony stimulating factors and transforming growth factor beta1 transiently stimulated the transcription of type VIII collagen mRNA. Maximal levels were reached after 2 h and 100 pg/ml granulocyte macrophage colony stimulating factor (4-fold), 1 U/ml macrophage colony stimulating factor (4.6-fold) and 1 ng/ml transforming growth factor beta1 (1.6-fold). While overnight treatment with colony stimulating factors stimulated the expression of transforming growth factor beta1 mRNA, short incubations did not influence or downregulate the transcription. In turn, treatment with transforming growth factor beta1 reduced the expression of granulocyte-macrophage and macrophage colony stimulating factor mRNA. The in vitro mRNA expression patterns were directly reflected in the distribution patterns found in intimal thickenings and advanced atherosclerotic lesions. This study demonstrates that colony stimulating factors and transforming growth factor beta1 modulate the transcription of type VIII collagen in vitro. Our data indicate a direct mechanism and exclude a pathway, which is mediated via the stimulation of transforming growth factor beta1 transcription. Our studies further support the hypothesis that colony stimulating factors in concert with transforming growth factor beta1 affect the collagenous composition of the extracellular vascular matrix. PMID- 10381275 TI - Characterization of apoptotic macrophages in atheromatous tissue of humans and heritable hyperlipidemic rabbits. AB - Apoptotic macrophages are regularly found in atherosclerotic plaques indicating programmed cell death as one of their regulatory controls. The objective of this study was to characterize in more detail apoptotic macrophages in atherosclerotic lesions of humans and heritable hyperlipidemic (HHL) rabbits. Macrophages were immunohistochemically analyzed using antibodies directed against alphaMbeta2 integrins (CD11b/CD18), CD44, major histocompatibility complex (MHC) class I and II, inducible nitric oxide synthase (iNOS), manganese superoxide dismutase (MnSOD), tumor necrosis factor alpha (TNFalpha), p53, c-jun/AP-1 and rabbit macrophages (RAM-11) and the TUNEL (TdT-mediated dUTP nick end labeling) technique. Colocalization studies of human atherosclerotic carotid and aortic tissue showed apoptotic plaque macrophages also being MnSOD-, alphaMbeta2 integrin-, CD44-, MHC class I- and II-, iNOS-, TNFalpha- and p53-immunoreactive. Similar results occurred in atherosclerotic aortas of HHL rabbits. Computer assisted morphometric analyses revealed a positive correlation of the area density of MnSOD-immunoreactive macrophages with those of alphaMbeta2-integrin- and CD44-immunoreactive ones, but not with those of MHC class I- and II- as well as of RAM-11-immunoreactive macrophages. We conclude that apoptotic macrophages located in atherosclerotic vessel wall are activated, antigen-presenting, integrin-expressing and oxidatively stressed cells. Since all these processes have been demonstrated to cause apoptosis of macrophages in vitro, we propose their potency accelerates the susceptibility of the macrophages to programmed cell death in atherosclerotic lesions. PMID- 10381276 TI - N(epsilon)-(carboxymethyl)lysine in atherosclerotic vascular lesions as a marker for local oxidative stress. AB - Previous studies suggested that N(epsilon)-(carboxymethyl)lysine (CML), as the major product of oxidative degradation of glycated proteins and unsaturated fatty acids, represents an integrative biomarker for oxidative stress. In the present study, the level of CML in morphologically normal as well as atherosclerotic vessel walls are immunohistochemically analyzed and the in vitro formation of CML determined from glycoxidation and lipid peroxidation processes. The analysis revealed negative staining results in normal arterial walls of fetal, juvenile and young adult origin. A minor positive staining was seen in normal arteries from adults between 40 and 60 years of age with a rise in the CML-staining further increasing with rising individual age. This staining was mainly restricted to the intimal extracellular matrix and there was no intracellular staining. In arteriosclerotic vessels, in contrast, the extracellular CML staining was significantly increased by approximately 3-fold also affecting the vascular media and adventitia. A strong intracellular staining was seen in macrophages. The degree of CML-staining correlated with the extent of the atherosclerotic changes. The in vitro studies showed a slow formation of CML of glycated proteins under aerobic conditions. No CML was formed under anaerobic conditions. Unsaturated fatty acids revealed a much faster formation of CML which reached high levels. The addition of vitamin E did not substantially suppress the CML-formation. The data suggest that the endogenous biomarker CML for oxidative stress accumulates slowly in normal arterial walls. This process is significantly increased in atherosclerosis. While the accumulation of CML in the extracellular matrix seemed to be the result of ongoing glycoxidation, the significant intracellular CML-formation in macrophages may have resulted from lipid peroxidation. PMID- 10381277 TI - Nitric oxide inhibits proliferation of human endothelial cells via a mechanism independent of cGMP. AB - Endothelial cell-derived nitric oxide (NO) has been suggested to inhibit smooth muscle cell proliferation, resulting in the reduction of intimal hyperplasia during atherogenesis. The present study investigates the role of NO from exogenous and endogenous sources on the proliferation of human umbilical vein endothelial cells (HUVEC) and human coronary artery endothelial cells (CAEC). Three different NO-generating compounds [sodium nitroprusside (SNP), S-nitroso glutathione (GSNO) and S-nitroso-acetylpenicillamine (SNAP)] were found to inhibit endothelial cell proliferation measured with three independent methods (cell counting, [3H]thymidine incorporation, DNA histograms) with significant inhibition occurring at concentrations > or = 100 microM. Growth-inhibiting effects were observed after long-term treatment (18-96 h) as well as after short stimulation with NO donors (10 min with a subsequent NO donor-free culture period of 18 h) and were comparable in culture medium (20% serum, growth factor supplementation) and serum-deficient medium (1% serum). The NO donor effects were mediated by the release of NO as they were prevented by NO scavenging. Superoxide dismutase (SOD) was found not to interfere with these effects suggesting that peroxynitrite formation was unlikely to be involved. 1H-[l,2,4]Oxadiazolo[4,3, alpha]quinoxalin-1-one (ODQ), a specific inhibitor of the soluble guanylate cyclase, was observed not to alter the antiproliferative effects of NO donors although it completely prevented NO-mediated increase of cyclic guanosine 3',5' monophosphate (cGMP), suggesting that the NO-induced growth inhibition was not mediated by cGMP. Furthermore, inhibition of endogenous NO production by N-nitro L-arginine methylester (L-NAME) did not affect endothelial cell growth regardless of using serum plus growth factor supplement, growth factor supplement alone, or thrombin to stimulate proliferation. We suggest that constitutively synthesized NO may not regulate endothelial cell proliferation whereas the growth-inhibiting NO effects may occur when an inducible NO synthase associated with a persistently high NO production is expressed in the atherosclerotic vessel wall. PMID- 10381278 TI - Physicochemical binding properties of the proteoglycan receptor for serum lipoproteins. AB - Proteoheparan sulfate can be adsorbed to a methylated silica surface in a monomolecular layer via its transmembrane hydrophobic protein core domain. Due to electrostatic repulsion, its anionic polysugar side chains are stretched out into the blood substitute solution representing a co-receptor for specific lipoprotein binding through basic amino acid-rich residues within their apolipoproteins. The binding process was studied by ellipsometric techniques showing that oxLDL had a deleterious effect on heparan sulfate proteoglycan binding and conformation. Ca2+ binding to and storage on the proteoheparan sulfate/LDL compound formed a 'heterotrimeric' HS-PG/LDL/Ca2+ complex of high stability, aggregability and deposit coating. On the other hand, HDL bound to heparan sulfate proteoglycan protected against LDL docking and completely suppressed calcification of the proteoglycan/lipoprotein complex. PMID- 10381279 TI - Oxidizability of low density lipoprotein and total antioxidative capacity of plasma are differently altered during induction and regression of hypercholesterolemia in rabbits. AB - Oxidizability of isolated low density lipoprotein (LDL) and total antioxidative capacity of plasma were measured in rabbits fed for 6 weeks a cholesterol-rich diet and for further 34 weeks a normal diet. Whereas the time to induce copper ion-mediated lipid peroxidation in LDL was prolonged during hypercholesterolemia, total antioxidative capacity as determined by a radical-trapping assay was increased at 6 weeks, but decreased during the time when the plasma cholesterol levels declined slowly to normal. Since aortic plaque progression was continued also during the first 15 weeks of normal diet, increased atherogenicity of hypercholesterolemia might be better reflected by the antioxidant capacity of plasma rather than by oxidation of isolated LDL. PMID- 10381280 TI - Analysis of secretory group II phospholipase A2 expression in human aortic tissue in dependence on the degree of atherosclerosis. AB - Secretory non-pancreatic (group II) phospholipase A2 (sPLA2) releases precursors of important mediators of inflammation from phospholipids. Based on the inflammatory character of atherosclerosis we previously described the identification of sPLA2 in human atherosclerotic plaques. In vitro studies on lipoproteins have shown that sPLA2 is able to favour the formation of foam-like cells representing a typical feature of early atherosclerotic lesions. In the present study the expression of sPLA2 in relation to the degree of atherosclerosis was investigated. Aortic tissue samples of 25 autopsy cases ranging in age from 1 to 77 years were taken from 2 cm above the heart and 3 cm below the renal arteries. The material was classified regarding the degree of atherosclerotic changes based on staining with haemalaun and eosine as well as on staining according to Goldner. Furthermore, immunohistochemical procedures detecting sPLA2, macrophages and smooth muscle cells were performed. The study has shown that in the abdominal aorta the enzyme was present in all advanced atherosclerotic lesions, but only in some preatheromas and precursors of atherosclerosis. However, this correlation did not occur in the thoracic aorta, where sPLA2-positive results showed a similar frequency in all degrees of atherosclerotic lesion. The enzyme was found in all three layers of the vessel wall without significant differences. Round cells, scarcely smooth muscle cells and endothelial cells were identified as sPLA2-positive. However, these data do not allow a conclusion as to which type of cell is responsible for the secretion of sPLA2. In summary, the correlation between the expression of this enzyme and the degree of atherosclerosis underlines the possible importance of sPLA2 in atherogenesis. PMID- 10381281 TI - Novel reversible, irreversible and fluorescent inhibitors of platelet-activating factor acetylhydrolase as mechanistic probes. AB - Phosphatidylcholines (1-O-alcoxy-2-amino-2-desoxy-phosphocholines and 1-pyrene labeled analogs) were synthesized and used to examine interactions with recombinant human PAF-acetylhydrolase (PAF-AH), an enzyme purified from plasma, and with macrophage-like U937 cells. Novel phosphatidylcholines containing a sn-2 carbamoylester group such as 1-O-hexadecyl-2-desoxy-2-amino-methylcarbamoyl-2 methyl-rac-glycer o-3-phosphocholine 11 were found to act as site-specific irreversible enzyme inhibitors with Ki-values up to 83 (K(irev)) and 177 (Ki(inact)) microm. The compounds exhibit only marginal inhibition of Ca2+ dependent phospholipases. Kinetic data show that phosphocholines carrying a terminal sn-1-pyrene moiety inhibit PAF-AH activity with an effectivity similar to analogs with an aliphatic chain. 1-O-Decyloxy-[10-(4-pyrenyl)-butoxy]-2-desoxy 2-amino-carbamoyl-me thyl-rac(-glycero-3-phosphocholine 13 could be used for enzyme labeling and to demonstrate an inhibitor-enzyme stoichiometry of 0.7:1. At 8 degrees C, the compound accumulated in the membranes of U937 cells, at 37 degrees C it was internalized into intracellular compartments. Structure activity studies in a mixed micelle assay indicated that the inhibition power of reversible and irreversible inhibitors increases along with the (sn)-1-chain length similar to the structure-dependent binding of ether phospholipids to the PAF-receptor. Unlike the situation at the (sn)-1-position, increasing chain length at the sn-2-position, or an alkyl branching of the glycerol backbone significantly reduced the inhibitory potency. PMID- 10381283 TI - Diminished susceptibility to in vitro oxidation of low-density lipoproteins in hypercholesterolemia: key role of alpha-tocopherol content. AB - Oxidizability of low-density lipoproteins (LDL) in hypercholesterolemia is of clinical relevance, but previous studies revealed diverging results. Therefore, we studied ex vivo oxidation of LDL in plasma samples of 57 hypercholesterolemic and 20 normocholesterolemic volunteers. LDL were isolated by ultracentrifugation and analyzed for lipids and alpha-tocopherol. The formation of conjugated dienes, lipoperoxides and malondialdehyde was measured at 0.1 micromol/l LDL, 3.2 micromol/l CuSO4. We found prolongation of the lagtime (53.6/65.8/71.4 min) with tertiles (< or = 3.17/3.89/14.2 mmol/l) of LDL-cholesterol (LDL-C). Regression analysis revealed that the lagtime increased with the apparent concentrations of cholesterol (P = 0.003) and alpha-tocopherol (P = 0.001) in the oxidation assay. A multiple regression model with the apparent concentrations of alpha-tocopherol, triglycerides and cholesterol explained 40% of the variation in lagtime. The close relationship between plasma concentrations of LDL-C and LDL-alpha tocopherol (P = 0.002) indicated that LDL contained more of this antioxidant in hypercholesterolemia. This might provide an explanation for the positive relationship between lagtime and LDL-C. The latter was independent of whether LDL C or LDL-protein was chosen for standardization of the oxidation assay. The formation of conjugated dienes (P = 0.000), lipoperoxides (P = 0.038) and malondialdehyde (P = 0.001) increased with the cholesterol level in the assay. This may be due to the increased load of LDL with cholesterol esters as a substrate for oxidation in hypercholesterolemia. Our data do not support the opinion that hypercholesterolemia is characterized by increased susceptibility of LDL to oxidation. PMID- 10381282 TI - Hyperhomocysteinemia in high-aged subjects: relation of B-vitamins, folic acid, renal function and the methylenetetrahydrofolate reductase mutation. AB - Moderate hyperhomocysteinemia is an atherogenic risk factor and plays an important role in geriatrics. Here, we have investigated the role of hyperhomocysteinemia in two elderly groups: 104 longeval subjects of 85-102 years, 100 seniors aged 65-75 years, and 75 controls of 19-60 years. Elevated homocysteine levels were found in 58% of longeval subjects in comparison with 32% in seniors. The homocysteine level in serum correlated positively with age as well as serum creatinine, and inversely with serum folate, but there was no correlation with serum B-vitamins. The frequency of vitamin B deficiency in serum of longeval subjects compared to seniors was as follows: vitamin B6 43% vs. 22%, vitamin B12 20% vs. 8%, and folic acid 1% in both groups. Increased serum creatinine levels (> 1.1 mg/dl) were found in 63% of the longeval subjects and in 32% of seniors. The 677-missense mutation in the methylenetetrahydrofolate reductase (MTHFR) gene, responsible for moderate homocysteine elevation, was found in 35, 37 and 27% of alleles in longeval persons, senior subjects and younger controls, respectively, showing no significant difference in frequency distributions of the MTHFR gene mutation. It can be concluded that hyperhomocysteinemia is very common with increased age. Its importance as an atherogenic risk factor with advanced age has to be clarified. PMID- 10381284 TI - Alpha-tocopherol down-regulates scavenger receptor activity in macrophages. AB - The aim of this study was to investigate the effect of alpha-tocopherol on scavenger receptor (SR) activity, SR class A (SR-A) mRNA expression and transcriptional regulation in macrophages. Scavenger receptor activity was determined quantitatively by uptake of DiI-acLDL (1,1'-dioctadecyl-3,3,3',3' tetramethylindocarbocyanine perchlorate-labeled acetylated low-density lipoprotein) in rabbit peritoneal macrophages and human monocytes/macrophages in the presence and absence of different tocopherol homologues. SR-A mRNA expression was determined by Northern blotting, the activity of the transcription factor activator protein-1 (AP-1) by electrophoretic mobility shift assay. We could demonstrate that alpha-tocopherol down-regulates scavenger receptor activity in macrophages in a dose-dependent manner. Scavenger receptor activity was reduced by 13, 16, 18 and 24% in the presence of 1, 5, 10 and 50 microM alpha-tocopherol, respectively. This effect was associated with a reduced SR-A mRNA expression and activity of AP-1 binding transcription factors in the presence of alpha tocopherol. The activity of scavenger receptors in human monocyte derived macrophages incubated with 100 microM alpha-tocopherol for 15 days was reduced up to 60%. Interestingly, gamma-tocopherol, which is a homologue of alpha-tocopherol with a comparable antioxidative capacity, showed only a weak suppression of SR activity, SR-A expression and AP-1 activity. Our observations point to the conclusion that the reduction of SR-A expression and activity in presence of alpha-tocopherol is possibly related to its direct action on cell signaling. PMID- 10381285 TI - Lipophilic antioxidants in blood plasma as markers of atherosclerosis: the role of alpha-carotene and gamma-tocopherol. AB - Oxidative theory of atherosclerosis implies that plasma levels of lipophilic antioxidants might serve as indicators of lipoprotein oxidation in the arterial wall and as markers of the development of atherosclerosis. However, it is unknown whether the measurement of plasma antioxidants is able to reflect atherogenesis or its risk. In order to assess whether the levels of lipophilic antioxidants in human plasma can discriminate between subjects with and without atherosclerosis, we measured the lipophilic antioxidants alpha-tocopherol, gamma-tocopherol, alpha carotene, beta-carotene and ubiquinol-10 in plasma of 34 patients with coronary heart disease (CHD) and in 40 control subjects. We found that alpha-carotene and gamma-tocopherol were significantly lower in plasma of CHD patients compared to controls. This decrease was significantly independent of whether the antioxidants were expressed as its absolute amounts in plasma (P < 0.001 for alpha-carotene, and P = 0.001 for gamma-tocopherol) or normalized to plasma lipids (P < 0.001 for both). In contrast, beta-carotene was only lower in plasma of CHD patients in comparison to controls, when normalized to the lipids (P = 0.02). Independent contributions of different parameters to the variation in these plasma antioxidants were estimated using multiple regression approach. The analysis showed that both the decrease in alpha-carotene and the decrease in gamma tocopherol were significantly associated only with the presence of CHD (P < 0.001 for each regression), while the decrease in beta-carotene was significantly related to the presence of hyperlipidaemia (P < 0.001). In striking contrast, no decrease in plasma alpha-tocopherol and ubiquinol-10 was detected in the patient group independently of how these antioxidants were expressed. These data suggest that plasma levels of alpha-carotene and gamma-tocopherol may represent markers of atherosclerosis in humans. Measuring these antioxidants may be of clinical importance as a practical approach to assess atherogenesis and/or its risk. PMID- 10381286 TI - A human arterial organ culture model of postangioplasty restenosis: results up to 56 days after ballooning. AB - BACKGROUND: Restenosis is a reparative process that is activated in response to injury induced by angioplasty. Despite numerous experimental models of restenosis the number of human arterial organ culture systems is very limited and long-term experiences do not exist. METHODS AND RESULTS: During routine nephrectomies parts of the renal arteries of 88 patients were extracted, 47 were suitable for organ culture preparations. Sections were made at 3 mm intervals perpendicular to the vessel wall axis. The arterial segments were treated with 3 mm standard balloon catheters (Medtronic 14K2030E) for 60 s with 3, 6, 9, and 12 bar. After angioplasty, the organ segments were cultured in a mixture of Waymouth's MB 752/1 and Ham F-12, supplemented with 15% fetal calf serum. After 0, 4, 14, 21, 28, and 56 days the organ cultures were fixed in 4% para-formaldehyde and embedded in paraffin. After staining with a modified elastica-van Gieson technique the intimal wall thickening was analyzed with a computerized morphometric system. For the identification of smooth muscle cells (SMC) a monoclonal antibody against smooth muscle alpha-actin was used. Endothelial cells were identified using an anti-human von Willebrand factor. To determine the number of cells undergoing DNA synthesis, bromodeoxyuridine (BrdU), a thymidine analogue, was added to the culture media 18 h prior to fixation. BrdU was detected with a monoclonal antibody, as secondary antibody a biotinylated horse-anti-mouse antibody was used. After 14, 21, and 28 days in culture BrdU-positive cells were detected in the neointima of the organ cultures, indicating mitotic activity in this area. After 28 and 56 days in culture a clear increase of neointimal thickening was found in the morphometric analysis. By positive reaction with antibodies against smooth muscle alpha-actin these cells were partly identified as SMC. CONCLUSIONS: The organ culture model offers opportunities for in vitro investigations of postangioplasty restenosis. The data emphasize the importance of a relatively late proliferative response of SMC in the human arterial organ culture model. PMID- 10381287 TI - Neointimal growth can be influenced by local adventitial gene manipulation via a needle injection catheter. AB - Revascularization by percutaneous transluminal coronary angioplasty is limited in the long-term by restenosis, which is luminal renarrowing in the first 6 months after the procedure. Smooth muscle cell proliferation is thought to be an important factor in restenosis; this leads to neointima formation and arterial lumen narrowing. Local therapy delivered perivascularly may have an effect on events in the neointima and reduce restenosis. The effect of delivering expression vector plasmids for senescent cell-derived inhibitor SDI-1, which regulates cell proliferation, and its antisense, into the perivascular tissue of injured arteries was investigated in a porcine arterial injury model using a needle injection catheter. Transfection efficiency, biological effect and plasmid dissemination were evaluated in arterial and organ tissue sections between 2 days and 4 months. A limited number of adventitial, medial and neointimal cells were transfected up to 4 months. sdi gene transfer did not result in a change in neointima. Transfer of antisense sdi resulted in an increase in neointima after 3 weeks. No DNA plasmid was detected in control tissues. Liposomally-mediated adventitial local gene delivery is feasible and safe using the needle injection catheter in a porcine model. A limited number of cells was transfected, with expression of transfected genes up to 4 months after delivery. A transient biological effect with increased neointima was observed after delivery of the antisense sdi gene. PMID- 10381288 TI - Correlation between coronary morphology and molecular markers of fibrinolysis in unstable angina pectoris. AB - BACKGROUND: In acute coronary syndromes, marked alterations of coagulation and fibrinolysis have been observed, but no data are available concerning a possible relation to coronary stenosis morphology. METHODS: Thirty one patients with unstable angina pectoris were included. Culprit stenosis morphology judged from coronary angiography was graded using the modified ACC/AHA classification. Molecular and functional markers of hemostasis and fibrinolysis were determined from venous plasma samples obtained at admission. RESULTS: Patients with unstable angina pectoris had a moderate procoagulant state, especially a contact phase activation compared with age-matched controls (factor XII 93.9 +/- 5.6 vs 112.8 +/- 5.4%; P < 0.05; high molecular weight kininogen 55.3 +/- 5.4 vs 86.1 +/- 6.5%; P < 0.01). Thrombin-antithrombin (TAT) was not significantly elevated (7.6 +/- 1.9 vs 4.0 +/- 0.5 microg/l). Elevated plasminogen activator mass concentration (16.6 +/- 2.1 vs 5.4 +/- 0.6 ng/ml; P < 0.01) and plasminogen activator inhibitor (PAI) activity (9.9 +/- 3.0 vs 5.6 +/- 3.0 AU/ml; P < 0.05) indicated an alteration of the fibrinolysis. Complexity of coronary stenosis was positively correlated with tissue-type plasminogen activator (TPA) mass concentration (P < 0.01) and PAI activity (P < 0.05). No association was found to markers of a hypercoagulative state. CONCLUSION: These findings indicate a relation between alterations of the fibrinolytic system and coronary morphology, whereas the acute changes of coagulation occur independently of culprit stenosis complexity. PMID- 10381289 TI - Low density lipoprotein apolipoprotein B metabolism: comparison of two methods to establish kinetic parameters. AB - Elevated plasma cholesterol concentrations represent a major risk factor for coronary artery disease (CAD). Low density lipoprotein (LDL)-apolipoprotein (apo) B plays a key role in this process. Metabolic parameters of LDL-apoB such as fractional catabolic rate (FCR) and production rate help to understand the underlying pathomechanisms of elevated LDL-apoB and are usually determined with tracer studies (gold-standard). However, these parameters can also be calculated from the rebound of plasma LDL-apoB concentration following a perturbation such as apheresis, if it is assumed that the perturbation itself does not affect metabolic parameters. LDL-apoB metabolism was determined using two methods in eight hyperlipoproteinemic patients (47 +/- 15 years, body mass index (BMI) 27.5 +/- 4.1 kg/m): (a) by endogenous labeling using D3-leucine (bolus 5 mg/kg) as tracer and multicompartmental modeling; and (b) by fitting a monoexponential equation to LDL-apoB rebound concentration data following apheresis. LDL-apoB metabolic parameters determined using the two methods (mean +/- S.D.; FCR-tracer: 0.18 +/- 0.07 per day, FCR-rebound: 0.27 +/- 0.25 per day; production-tracer: 12.0 +/- 3.9 mg/kg per day; production-rebound: 15.2 +/- 8.0 mg/kg per day) were not correlated, were not concordant (intraclass correlation coefficient), and were not significantly different. Furthermore, only in five of the eight patients the rebound analysis predicted LDL-apoB steady-state concentrations that were within 20% of observed steady-state concentrations. These results indicate that parameters derived from LDL-apoB mass rebound following apheresis cannot be used as a surrogate for parameters established with tracer methodology probably because the assumption that apheresis does not affect metabolic parameters of LDL apoB may not be true in all patients. PMID- 10381290 TI - Lifibrol enhances the low density lipoprotein apolipoprotein B-100 turnover in patients with hypercholesterolemia and mixed hyperlipidemia. AB - Lifibrol (4-(4'-tert-butylphenyl)-1-(4'carboxyphenoxy)-2-butanol), a new hypocholesterolemic drug, effectively reduces total cholesterol (CH), low density lipoprotein (LDL)-CH, and apolipoprotein (apo) B in experimental animals and in humans. The impact of Lifibrol on the metabolism of apoB-100 containing lipoproteins in patients with hyperlipoproteinemia using endogenous labeling with stable isotopes is examined. Kinetic studies were performed in four male hypercholesterolemic individuals (type IIa) before and on treatment with 450 mg of Lifibrol daily for 4 weeks, and in five male individuals suffering from mixed hyperlipidemia (type IIb) before and on therapy for 12 weeks. Kinetic parameters were estimated by multicompartmental modeling. Lifibrol therapy reduced total CH by 16% (P = 0.012) in all patients, increased triglycerides (TG) by 11% (not significant) in type IIa patients and decreased TG by 34% (P = 0.059) in type IIb patients. During Lifibrol therapy, LDL apoB-100 concentrations decreased by 19% (P = 0.011) in all patients. The decrease in LDL apoB concentrations with Lifibrol therapy was due to an overall increase (75%, P = 0.006) of the fractional catabolic rates (FCR) of LDL apoB. This increase was partially attenuated by a 33% increase in LDL apoB production rate (PR) (P = 0.041). The overall production of apoB increased only slightly. Our data suggest that the major mechanism by which Lifibrol lowers LDL-CH is an increase in receptor mediated catabolism of LDL rather than a decrease in hepatic apoB production. PMID- 10381291 TI - Metabolic basis of high density lipoproteins and apolipoprotein A-I increase by HMG-CoA reductase inhibition in healthy subjects and a patient with coronary artery disease. AB - HMG-CoA reductase inhibitors, such as pravastatin, are widely used as lipid lowering drugs in hypercholesterolemia. Pravastatin does not only reduce the atherogenic low density lipoprotein (LDL)-cholesterol, but is also increasing high density lipoprotein (HDL)-cholesterol. However, the mechanism leading to an increase of HDL are unclear. Therefore, the effects of pravastatin on the in vivo kinetics of apolipoprotein (apo) A-I were studied in six normolipidemic subjects and in a patient with coronary artery disease (CAD) utilizing stable isotope tracer techniques. Two turnover studies were performed. The first turnover study was carried out before any drug treatment, the second study after 6 weeks of 40 mg pravastatin/day. Three times deuterium labeled L-leucine (3D-leucine) was given as a primed bolus constant infusion (bolus: 1340 microg/kg; infusion: 22 microg/kg per h), and tracer uptake into HDL apoA-I was determined by gas chromatography (GC)-mass-spectrometry (MS). In the healthy subjects HDL cholesterol increased by 13% and apoA-I increased by 12% under pravastatin treatment. The HDL in the CAD patient decreased by 3% and apoA-I increased by 2%. Prior to drug treatment the mean apoA-I fractional synthetic rate (FSR) was 0.194 per day (S.D. +/- 0.02) and apoA-I production rate (PR) was 10.8 mg/kg per day (S.D. +/- 2.1). The CAD patient had a FSR of 0.219 per day and a PR of 10.6 mg/kg per day. After treatment with pravastatin the mean apoA-I FSR was 0.204 per day (S.D. +/- 0.02) and apoA-I PR was 12.5 mg/kg per day (S.D. +/- 1.5) in the healthy subjects. Despite only minor changes of HDL and apoA-I in the CAD patient, there were significant changes of FSR (0.267 per day) and PR (13.1 mg/kg per day) with pravastatin treatment. The in vivo kinetic data demonstrate an increased FSR of apoA-I. The increase in apoA-I is due to an increased PR of apoA I. This study demonstrates increased production of HDL apoA-I as the metabolic cause of the increase in HDL and apoA-I levels under inhibition of HMG-CoA reductase in man. PMID- 10381292 TI - Prevalence of risk factors of coronary heart disease in Turks living in Germany: The Giessen Study. AB - Turkish people represent the majority of immigrants in Germany. Even though a high proportion of Turks has been living in Germany since about 20 years, little is known about risk factors of coronary heart disease (CHD) in this population. In this study a sample of 325 male and 155 female Turks are investigated, who voluntarily underwent a health check-up in Germany. Data about the presence of CHD, risk factors and blood parameters were collected. Mean residence time was 21 and 17 years (males/females). A low percentage of female participants was observed compared to the general Turkish population in Germany. Age adjusted prevalence of CHD reached 9.5% in males and 6.7% in females, respectively. Dyslipoproteinemia (DLP) showed the highest prevalence of all risk factors investigated in both genders. Total cholesterol (TC) levels were comparable to those of other western countries and remarkably higher than reported for the population in Turkey. Besides this, low high density lipoprotein-cholesterol (HDL C) and apolipoprotein A-I (ApoA-I) levels could be found in the majority of the sample. The highest odds ratios for CHD were estimated for stress and hypertension in males and obesity in females. It is concluded that Turkish immigrants in Germany showed an assimilation of lipid pattern to western populations. However, reasons for low HDL-C levels remain unclear. Changes in the lipid metabolism chiefly seem to contribute to the risk factor pattern of Turkish immigrants in Germany. PMID- 10381293 TI - Obesity, mortality and cardiovascular disease in the Munster Heart Study (PROCAM). AB - In the Munster Heart Study (PROCAM), 16,288 men aged 40.6 +/- 11.3 years (mean +/ S.D.) and 7328 women aged 36.0 +/- 12.3 years were enrolled between 1979 and 1991. Mean body mass indices (BMIs) were 25.6 +/- 3.3 and 23.8 +/- 4.1 kg/m2 in men and women, respectively. There was a graded and continuous positive interaction in both men and women between BMI, age and serum total cholesterol, low density lipoprotein (LDL) cholesterol, and blood pressure (both systolic and diastolic). High density lipoprotein (HDL) cholesterol tended to increase with age, but decreased in graded fashion with increases in BMI in both sexes. Triglyceride increased with BMI in both sexes and with age in women, but decreased in the older age groups of overweight and obese men. Though fasting blood glucose increased with age and BMI in both sexes, the increase was more marked in women. Among the 10,856 men aged 36-65 years at study entry, 313 deaths occurred within a follow-up period of 7.1 +/- 2.4 years. Among these men, increased mortality was seen at high BMIs in both smokers and non-smokers and was caused by coronary heart disease (CHD). Increased mortality at low BMI was seen in smokers but not in non-smokers and was due to an increase in cancer deaths. The BMI-associated increase in CHD death was completely accounted for by the factors contained in the Munster Heart Study (PROCAM) risk algorithm, indicating that the effect of overweight and obesity on CHD is mediated via other risk factors. PMID- 10381294 TI - Spontaneous platelet aggregation as a predictive risk factor for vascular occlusions in healthy volunteers? Results of the HAPARG Study. Haemostatic parameters as risk factors in healthy volunteers. AB - The HAPARG Study (haemostatic parameters as risk factors in healthy volunteers) was performed in a subset of volunteers taking part in the MARISK Study (Mainzer Risikoindikatoren Studie fur die koronare Herzkrankheit) sponsored by the German Ministry of Research and started in 1984. A previous study (Yamanishi et al., Thromb Haemostas 1985;54:539-543) had shown that spontaneously enhanced platelet aggregation as measured with the PAT-III-test and higher fibrinogen concentrations are significant risk factors for new vascular occlusions in diabetic patients. It was the aim of the HAPARG Study to establish whether spontaneous platelet aggregation and other hemostatic variables are independent risk factors for vascular occlusions in healthy volunteers. Employees of a chemical/pharmaceutical company aged 40-65 years and personnel of the University of Mainz, aged 30-60 years were included in this prospective study. Besides anamnestic data such as on smoking, hypertension and diabetes, blood pressure, the ankle/arm Doppler-index and an ECG were recorded and serum cholesterol, HDL, LDL, triglycerides, uric acid and glucose were measured. Men (1884) and women (989) entered the study and were followed for 4-6 years. In the age group of 30 50 years, more women than men were included. During the observation period 53 vascular occlusions occurred (36 coronary and nine cerebral events and eight peripheral vascular occlusions). Only three of these endpoints occurred in women. Besides age (odds ratio = 1.7, P = 0.02) and gender as expected risk factors, the multivariate logistic stepwise regression analysis revealed smoking (odds ratio = 2.2, P = 0.008), lower HDL-levels (odds ratio = 2.2, P = 0.013), elevated diastolic blood pressure (odds ratio = 1.4. P = 0.004) followed by spontaneous platelet aggregation (odds ratio = 1.1, P = 0.037), and slightly elevated blood glucose (P = 0.0047) as significant risk factors for men. Higher fibrinogen levels missed significance in this analysis (P = 0.059). None of the other hemostatic parameters showed a significant correlation with the vascular events. To our knowledge, this has been the first prospective trial in a large population of healthy individuals in which a platelet function parameter has been studied together with other possible risk factors. Spontaneously enhanced platelet aggregation is probably an independent risk factor and, like elevated fibrinogen and other haemostatic variables, an indicator of an ongoing active atherosclerotic process. PMID- 10381295 TI - The significance of high levels of lipoprotein (a) compared with established risk factors in premature coronary artery disease: differences between men and women. AB - It was shown in a series of studies that increased lipoprotein (a) concentration is a strong and independent risk factor for coronary artery disease. The goal of this study was to determine the significance of elevated lipoprotein (a) levels for the existence and the early manifestation of coronary artery disease by systematically recording cardiovascular risk factors in diagnostic coronary angiographies in a larger group of patients, whereby particular attention was paid to sex-specific differences. In 1011 consecutive patients who underwent coronary angiography (731 men, 280 women, mean age 59 +/- 10 years), fasting blood samples were taken immediately before the angiographies to determine the levels of cholesterol, low density lipoprotein-, high density lipoprotein cholesterol, triglycerides and lipoprotein (a). In addition, further risk factors were qualitatively recorded. The data evaluation was carried out using the SPSSx software package univariately and multivariately with stepwise discriminant analysis. In 231 patients (144 men, 87 women) either no or only discrete coronary findings appeared, while in 780 cases (587 men, 193 women) coronary artery disease with stenoses > 50% were found. Women with coronary artery disease were significantly older than men and demonstrated higher lipoprotein levels. Women as well as men with coronary artery disease differed from healthy controls by having higher levels of lipoprotein (a) and other lipoproteins, lipoprotein (a) having the smallest error probability (P < 0.0005). The early manifestation of coronary artery disease (below the 18th age percentile) in men (< 50 years) was connected with significantly higher levels of cholesterol, triglycerides and lipoprotein (a), which emphasized their atherogenic significance in the general view. The most striking finding was that in young women (< 53 years), compared to older women with coronary artery disease--corresponding to the age-determined prevalence--significantly lower concentrations of cholesterol, triglycerides and lipoprotein (a) were found. Possible explanations include later manifestation of coronary artery disease, a steeper increase of the lipids with age, particularly of lipoprotein (a), but also a different valence of the risk factors in women. PMID- 10381296 TI - Postprandial plasma glucose is an independent risk factor for increased carotid intima-media thickness in non-diabetic individuals. AB - Postprandial (pp) hyperglycemia--frequently associated with an increase in cardiovascular risk factors--may be damaging for the endothelium. So far, little information exists how glucose, insulin and lipids may affect atherosclerosis in the pp state. Therefore, we evaluated the relationship of pp hyperglycemia, insulin secretion and coronary risk factors to intima-media thickness (IMT) in a non-diabetic risk population. In 403 subjects (147 males, 256 females), aged 40 70 years, in the majority relatives of index cases with type 2 diabetes--a 75 g oral glucose tolerance test was performed together with measurement of insulin fractions, various risk factors and IMT of the common carotid artery. We found a continuous rise of 2h pp insulin fractions along the quintiles of 2h pp plasma glucose. A significant increase of body mass index, waist to hip ratio, triglycerides and decrease of HDL-cholesterol was observed in the top quintile of 2h pp plasma glucose (8.24 > or = pp plasma glucose < 11.1 mmol/l). Albuminuria was significantly enhanced in the 5th quintile. In parallel, IMT was significantly increased in the 5th quintile versus the bottom quintile of 2 h and maximal glucose (range 11.7-15.3 mmol/l) postprandially. After age and sex adjustment pp glucose and C-peptide, total cholesterol, triglycerides and HDL cholesterol but not fasting plasma glucose were significantly correlated to IMT. In multivariate analysis age, male sex, pp plasma glucose, total and HDL cholesterol were found to be independent risk factors for increased IMT. In conclusion, our data in a non-diabetic European risk population show that the two top quintiles of pp plasma glucose are associated with a clustering of standard risk factors. Corresponding to this clustering of risk factors IMT was significantly increased in the top quintile of 2 h and maximal pp plasma glucose. These data show that pp hyperglycemia may exert a noxious impact on the arterial wall together with a cluster of anomalies typical for the metabolic syndrome. PMID- 10381297 TI - The antiatherosclerotic effect of Allium sativum. AB - In a randomized, double-blind, placebo-controlled clinical trial, the plaque volumes in both carotid and femoral arteries of 152 probationers were determined by B-mode ultrasound. Continuous intake of high-dose garlic powder dragees reduced significantly the increase in arteriosclerotic plaque volume by 5-18% or even effected a slight regression within the observational period of 48 months. Also the age-dependent representation of the plaque volume shows an increase between 50 and 80 years that is diminished under garlic treatment by 6-13% related to 4 years. It seems even more important that with garlic application the plaque volume in the whole collective remained practically constant within the age-span of 50-80 years. These results substantiated that not only a preventive but possibly also a curative role in arteriosclerosis therapy (plaque regression) may be ascribed to garlic remedies. PMID- 10381298 TI - A more mature phenotype of blood mononuclear phagocytes is induced by fluvastatin treatment in hypercholesterolemic patients with coronary heart disease. AB - Monocytes are recruited as the principal inflammatory cells into the atherosclerotic lesion. In a previous study we demonstrated that a low HDL cholesterol and the apo E4 allele are associated with an increased proportion of blood monocytes that are characterized by a high expression of Fcgamma-RIIIa (CD16), a dim expression of the lipopolysaccharide (LPS) receptor (CD14) and a high expression of beta1- and beta2-integrins (Rothe et al. Arterioscler Thromb Vasc Cell Biol 1996;16:1437-1447). In this study, 79 hypercholesterolemic patients were treated either with the HMG CoA reductase inhibitor fluvastatin in combination with diet or with placebo and diet in a double-blind and randomized multicenter study, and monitored for the potential effects on the phenotype of peripheral blood monocytes. At baseline, in the whole group of hypercholesterolemic patients the population size of these more mature monocytes (CD14dimCD16+) was positively correlated to triglyceride (P = 0.003) and total serum cholesterol levels (P = 0.012) confirming our previous study. Fluvastatin treatment for 52 weeks was associated with a 24.2% reduction in LDL-cholesterol (P < 0.001) as well as a 40.7% decrease in the expression density of CD14 on all monocytes (P = 0.027). A 24.5% decrease (P < 0.001) of the population of less differentiated CD14brightCD16- monocytes and an 83.1% increase (P = 0.029) of the population of more differentiated CD14dimCD16+ monocytes further confirmed this modification of the phenotype of peripheral blood monocytes. The positive pre study correlation of the CD14dimCD16+ monocyte subset to the serum cholesterol concentration, but inverse changes of both parameters under fluvastatin therapy, in conclusion indicate that fluvastatin exerts an as yet uncharacterized immunomodulatory effect on either monocyte maturation and differentiation, or extravasation which may also depend on the endothelial phenotype that is independent of the change in serum lipids. PMID- 10381299 TI - The effect of fluvastatin on cardiac events in patients with symptomatic coronary artery disease during one year of treatment. AB - The primary objective of the present study was to investigate the cholesterol lowering effect of fluvastatin on the incidence of cardiac events in hyperlipidaemic patients with symptomatic, clinically-diagnosed (exercise-ECG) coronary heart disease (CHD) during 1 year of treatment. Exercise tolerance, incidence of angina pectoris episodes, use of anti-anginal medication and intimal medial-thickness (IMT subgroup) of the A. carotis were secondary endpoints. In the double-blind trial a total of 365 male and female patients with stable symptomatic CHD and a low-density lipoprotein cholesterol (LDL-C) above 160 mg/dl on a lipid-lowering diet were randomised to fluvastatin 40 mg (o.a.d. or b.i.d.) or placebo for 1 year. Fluvastatin lowered total cholesterol by 17% and LDL-C by 27%. There was a significantly lower incidence of cardiac events (cardiac death, nonfatal myocardial infarction, unstable angina pectoris) in the fluvastatin group (3 events) as compared to the placebo group (10 events) (P < 0.05). Exercise tolerance improved and the incidence of angina pectoris episodes decreased in both groups, but more pronounced on fluvastatin (n.s.). Exercise-ECG discontinuation due to angina pectoris and ST-segment depression decreased in the fluvastatin group by 55.6 and 70.9%, respectively, and in the placebo group by 39.6 and 46.5% (n.s.). At baseline, a subgroup of 76 patients showed a mean IMT value of 0.73 mm which remained uninfluenced in the fluvastatin and the placebo groups. Fluvastatin was safe and well tolerated. In conclusion, patients with symptomatic CHD get cardiovascular benefit from lipid-lowering therapy with fluvastatin even during the first year of treatment. PMID- 10381300 TI - Bioaccumulation of mercury in various fish species from Orlik and Kamyk water reservoirs in the Czech Republic. AB - The study evaluated contamination of fish by mercury in two important Czech water reservoirs, Orlik and Kamyk, which receive water from the whole watershed of Otava and Vltava rivers. Contamination of these reservoirs reflects the situation in southern and western Bohemia. Six environmentally representative sites were sampled in the area of interest and the evaluation was based on regression models relating Hg concentrations in fish tissues to the age of fish. A total number of 170 analyzed individuals from 3 indicative species (Rutilus rutilus, Abramis brama, and Perca fluviatilis) allowed site-specific analyses of model residuals. The results of biological monitoring corresponded to the sediment analyses and suggested that Orlik reservoir receives a substantial part of the pollution from the Vltava river. Temporary oxygen deficiency and decreased value of pH were probably the key factors responsible for the increased exposure of fish to Hg in the Kamyk reservoir. An additional comparison of 286 analyzed individuals belonging to 11 fish species provided the following decreasing order for muscle contamination: typical predators/=9 months) to the replacement protein. MPS VI cats on shorter term ERT (3 months) showed high titers to 4-sulfatase and similar patterns of epitope reactivity, but lower affinity antibody reactivity, when compared to the latter cat. This study reports the linear amino acid sequence reactivity and nature of the immune response produced to 4-sulfatase before and after ERT. The monitoring of antibody production during replacement therapy is an important consideration for patient management, as high titer antibodies can affect the efficacy of therapy. PMID- 10381328 TI - Metachromatic leukodystrophy: subtype genotype/phenotype correlations and identification of novel missense mutations (P148L and P191T) causing the juvenile onset disease. AB - Metachromatic leukodystrophy (MLD) is a lysosomal storage disease resulting from the deficient activity of arylsulfatase A (ASA) and the accumulation of sulfatides. The disease is characterized by several subtypes, designated by age at onset: the late-infantile-, juvenile-, and adult-onset variants. Mutation analysis of genomic DNA from a proband with each variant was performed to identify and characterize their causative ASA mutations. Two sisters with the infantile-onset disease were homoallelic for the missense mutation D335V, a juvenile-onset proband was heteroallelic for two novel missense mutations, P148L and P191T, and an adult-onset patient was heteroallelic for the H397Y and P426L mutations. The novel mutations were not identified in 108 normal alleles indicating that these base substitutions were not common polymorphisms. To further characterize the mutant gene products, the mutant enzymes were partially purified from cultured fibroblasts and their molecular weights and charges were compared by immunoblotting following SDS-PAGE or isoelectric focusing (IEF). Normal fibroblast ASA had a single, broad band at 54 kDa. The enzyme from the late-infantile-onset patient had distinct bands of 36 and 78 kDa, but lacked the normal 54-kDa species. The juvenile- and adult-onset patients each had a faint band of 54 kDa and several other bands ranging from 29 to 64 kDa. IEF revealed several bands for the partially purified normal enzyme with a relatively narrow pH range around 4.0, whereas numerous bands with a wider range of isoelectric points were observed with the enzymes from the juvenile- and adult-onset fibroblasts. In contrast, the enzyme from the late-infantile-onset proband had four bands with more acidic isoelectric points, none corresponding to those of the normal enzyme. These results document changes in both size and charge of the mutant enzymes from patients with different mutations and MLD subtypes. PMID- 10381329 TI - Muscle-specific transcription factors in fibroblasts expressing the alpha striated tropomyosin 3' untranslated region. AB - The alpha-striated tropomyosin 3' untranslated region (TM UTR) promotes differentiation of fibroblasts into cells resembling skeletal muscle. To investigate the mechanism of this observation, RNA harvested from transfected primary fibroblasts was used for semiquantitative RT-PCR with primers specific for muscle transcription factors, showing that myoD and myogenin transcripts are detected in these cells, but that differentiation after TM UTR expression is independent of a detectable increase in these transcripts. Double immunofluorescent staining with antibodies to myoD family members and to titin confirms that muscle differentiation in TM UTR-transfected fibroblasts is independent of production of any transcription factor in this family. In contrast, the muscle transcription factor myocyte enhancer factor 2 (mef-2) is strongly expressed after transfection of fibroblasts with the TM UTR. The increase in mef-2 protein is due to an increase in the steady-state level of its mRNA, as shown by Northern analysis. The expression of p21 ordinarily observed in skeletal myogenesis before the expression of muscle-specific proteins is not seen in fibroblasts induced to differentiate by the TM UTR. These results demonstrate that post-transcriptional regulation of myoD family members is seen in fibroblasts, and that the TM UTR induces muscle differentiation independent of the myoD transcription factors and without expressing proteins characteristic of terminal withdrawal from the cell cycle. Finally, an increase in the steady-state level of mef-2 transcripts appears in the proximal pathway of myogenic activation in response to expression of the TM UTR. These results imply that fibroblasts can utilize an additional differentiation route upon TM UTR expression resulting in mature muscle other than that requiring myoD family members. PMID- 10381330 TI - Isolation and characterization of a novel transcript embedded within HIRA, a gene deleted in DiGeorge syndrome. AB - We have isolated a few cDNAs from different human tissues, transcribed from the first intron of HIRA, a gene deleted in the DiGeorge syndrome. These cDNAs are produced by an intronic gene (22k48) which is transcribed by the HIRA opposite strand and is itself arranged in exons and subjected to alternative splicing. The longest continuum cDNA sequence we obtained is 3.6 kb long and contains 3 different exons and 2 introns. 22k48 cDNA is composed of several tandemly arranged repeated elements (Alu, LINEs, CAn) surrounding a unique sequence. In situ hybridization showed the presence of 22k48 RNA in the cytoplasm of CNS and PNS neurons. 22k48 RNA is able to bind cytoplasmic proteins in the range of 45 to 60 kDa. 22k48 is a new member of the small group of genes that are transcribed but not translated, and its haploinsufficiency could contribute to the pathogenesis of the DiGeorge syndrome. PMID- 10381331 TI - Cl- and membrane potential dependence of amino acid transport across the rat renal brush border membrane. AB - The relative roles of the anion present and the membrane potential in the operation of each of the seven amino acid transport systems in the renal tubular brush border membrane were explored by manipulating transmembrane potential and chemical gradients across the membrane. The effect of various external anions with different permeabilities of the membrane and of valinomycin-generated K+ diffusion potential on Na+-coupled amino acid accumulation by rat renal brush border membrane vesicles was examined. Accumulation of all amino acids examined, except for cystine, was membrane potential dependent. The highest voltage dependence was observed for taurine (equivalent to glucose) and l-methionine. Addition of taurine uptake values obtained under each electrical gradient (inside negative) and a chemical gradient (100 mM NaCl out) condition yielded markedly lower values than under conditions where there was a combined electrochemical gradient. Cl- gradient rather than merely imposing a voltage gradient was a specific mediator of Na+-coupled transport of l-proline, taurine, l-glutamic acid, and glycine across the brush border membrane. Cl- gradient alone under Na+ equilibrated conditions could energize an overshoot of taurine accumulation by vesicles providing evidence that taurine is energetically activated by and coupled to Cl- transport. These data suggest that Na+-linked transport of most amino acids across the tubular luminal membrane is an electrogenic positive process and for proline, taurine, glutamic acid, and glycine, a Cl--requiring process. A negative intracellular potential combined with luminal chloride is required for optimal Na+-coupled transport of these amino acids across the luminal membrane of the proximal tubule. The coupling of Cl- to the transport of these osmoprotective amino acids may enhance their volume regulatory effect in kidney cells and other mammalian cells. PMID- 10381332 TI - Association of the serotonin transporter gene with serum cholesterol levels and heart disease. AB - In a study of a group of elderly athletes we observed an unexpected association between serum cholesterol levels and the HTTLPR insertion/deletion polymorphism of the promoter region of the serotonin transporter gene (HTT, SLC6A4). As a follow-up we examined the potential association of this polymorphism with cholesterol and triglyceride levels, or heart disease, in two other groups of subjects. We examined the possible association between cholesterol levels and heart disease and genotypes of the HTTLPR insertion/deletion polymorphism of the promoter region of the HTT gene, in three independent study populations ranging from 42 to 90 years of age. For subjects 55 to 70 years of age in Group 1, cholesterol levels were significantly greater in the LS heterozygotes than either LL or SS homozygotes, indicating a heterosis effect (P 70 years of age. While these studies are preliminary and exploratory, they are consistent with a relationship of the HTT gene in cholesterol levels and a risk for heart disease. Replication of these findings in independent, epidemiologically based studies is required. PMID- 10381333 TI - Assessment of the nutritional value of glycerol-1,2, 3-tris(methylsuccinate) in fed and starved rats. AB - The nutritional value of glycerol-1,2,3-tris(methylsuccinate), a novel ester of succinic acid with high insulinotropic efficiency both in vitro and in vivo, was assessed in both fed and starved rats. The infusion of the ester, given in a daily amount (1.2 micromol. g body wt-1) well in excess of what could result from its repeated intravenous administration as an insulinotropic agent in non-insulin dependent diabetes (0.07 micromol. g body wt-1 for each administration), failed to prevent the fall in body weight, liver and muscle glycogen contents, and plasma d-glucose or insulin concentration, as well as the increase in plasma free fatty acid and beta-hydroxybutyrate concentrations caused by starvation. The sole indications that the ester may serve, to a limited extent, as an alternative nutrient in starved rats consisted in a somewhat higher weight of both liver and paraovarian adipose tissue and somewhat higher activity of liver glucokinase in rats receiving the ester than in animals infused with saline. The low nutritional value of this ester thus answers the objection of its possible role as an extrapancreatic nutrient or gluconeogenic precursor in the perspective of its use as an insulinotropic tool in type 2 diabetes. PMID- 10381334 TI - Heterologous overexpression of human NEFA and studies on the two EF-hand calcium binding sites. AB - Human NEFA is an EF-hand, leucine zipper protein containing a signal sequence. To confirm the calcium binding capacity of NEFA, recombinant NEFA analogous to the mature protein and mutants with deletions in the EF-hand domain were expressed in Pichia pastoris and secreted into the culture medium at high yield. The calcium binding activity of each purified protein was measured by a modified equilibrium dialysis using the fluorescent Ca2+ indicator FURA-2 and atomic absorption spectroscopy. A stoichiometry of 2 mol Ca2+/mol NEFA was determined. The Ca2+ binding constants were resolved by intrinsic fluorescence spectroscopy. Fluorescence titration exhibited two classes of Ca2+ binding sites with Kd values of 0.08 microM and 0.2 microM. Circular dichroism (CD) spectroscopy showed an increase from 30 to 43% in the amount of alpha-helix in NEFA after addition of calcium ions. Limited proteolytic digestion indicated a Ca2+ dependent conformational change accompanied by an altered accessibility to the enzyme. PMID- 10381336 TI - Tetracyclines reduce Na+/K+ pump capacity in Calu-3 human airway cells. AB - We studied the effect of tetracyclines on the Na+/K+ pump activity in Calu-3, a human airway cell line. To estimate Na+/K+ pump capacity on the basolateral membrane, an ouabain-sensitive component of the short-circuit current (Isc) was measured in the presence of nystatin, an ionophore of Na+. The application of ouabain (1 mM) to the basolateral solution completely inhibited the Isc generated by adding nystatin (50 microM) to the apical solution. Tetracycline (TC), minocycline (MC), or demethylchlortetracycline (DC) at 0.5 mM applied to the apical but not to the basolateral solution also decreased the nystatin-induced Isc. Neither phlorizin- nor diphenylamine-2-carboxylic acid-sensitive Isc was affected by TC, MC, or DC. These results indicate that tetracyclines may permeate only through the apical membrane with the result that the Na+/K+ pump's capacity for Na+ extrusion should be suppressed without a decrease in Cl- transport. PMID- 10381335 TI - A G-CSF receptor-gyrase B fusion gene: A new type of molecular switch for expansion of genetically modified hematopoietic cells. AB - We have developed a novel system for expansion of transduced hematopoietic cells. This system involves "selective amplifier genes" encoding fusion proteins between the granulocyte colony-stimulating factor receptor (Gcr) and the estrogen receptor (Er). The GcrEr chimeric gene conferred estrogen-dependent growth ability on murine hematopoietic cells. Here, we constructed a modified "selective amplifier gene" to circumvent possible concerns with the Er/estrogen switching system. The bacterial gyrase B (Gyr) gene was fused to the Gcr gene, and the GcrGyr fusion construct was introduced into interleukin-3 (IL-3)-dependent Ba/F3 cells. The dimeric antibiotic coumermycin induced IL-3-independent growth in Ba/F3 cells expressing GcrGyr. This stimulatory effect was antagonized by an excess amount of novobiocin, a monomeric form of coumermycin. These results suggest the feasibility of using Gyr as a molecular switch to regulate a growth signal in hematopoietic cells. PMID- 10381337 TI - Heterogeneous nuclear ribonucleoprotein A2 interacts with protein kinase CK2. AB - The catalytic subunit of protein kinase CK2 (CK2alpha) was found associated with heterogeneous nuclear ribonucleoprotein particles (hnRNPs) that contain the core proteins A2 and C1-C2. High levels of CK2 activity were also detected in these complexes. Phosphopeptide patterns of hnRNP A2 phosphorylated in vivo and in vitro by protein kinase CK2 were similar, suggesting that this kinase can phosphorylate hnRNPA2 in vivo. Binding experiments using human recombinant hnRNP A2, free human recombinant CK2alpha or CK2beta subunits, reconstituted CK2 holoenzyme and purified native rat liver CK2 indicated that hnRNP A2 associated with both catalytic and regulatory CK2 subunits, and that the interaction was independent of the presence of RNA. However, the capability of hnRNP A2 to bind to CK2 holoenzyme was lower than its binding to the isolated subunits. These data indicate that the association of CK2alpha with CK2beta interferes with the subsequent binding of hnRNP A2. HnRNP A2 inhibited the autophosphorylation of CK2beta. This effect was stronger with reconstituted human recombinant CK2 than with purified native rat liver CK2. PMID- 10381338 TI - Quantitative analysis of expression of mouse sialyltransferase genes by competitive PCR. AB - The present paper describes a rapid and systematic method for semi-quantitative analysis of the expression of sialyltransferase genes. So far, fifteen sialyltransferase cDNAs have been cloned from mice. Most of these genes are expressed in developmental stage-dependent and/or tissue-specific manners, and the expression levels of some of them are too low to detect on Northern blot analysis. To resolve how each sialyltransferase contribute to synthesize sialylglycoconjugates, it is necessary to establish the method for quantification of gene expression levels of these fifteen sialyltransferases. Therefore, we developed a competitive PCR-based method for analyzing the quantitative relationship of the gene expression of fifteen sialyltransferases. Using this method, we can investigate the levels of gene expression of sialyltransferases in various cell lines and various tissues of mice, and can accurately determine their expression levels. PMID- 10381339 TI - Preliminary characterization of Yor180Cp: identification of a novel peroxisomal protein of saccharomyces cerevisiae involved in fatty acid metabolism. AB - Here we report the preliminary characterization of Yor180Cp, a novel peroxisomal protein involved in fatty acid metabolism in the yeast Saccharomyces cerevisiae. A computer-based screen identified Yor180Cp as a putative peroxisomal protein, and Yor180Cp targeted GFP to peroxisomes in a PEX8-dependent manner. Yor180Cp was also detected by mass spectrometric analysis of an HPLC-separated extract of yeast peroxisomal matrix proteins. YOR180C is upregulated during growth on oleic acid, and deletion of YOR180C from the yeast genome resulted in a mild but significant growth defect on oleic acid, indicating a role for Yor180Cp in fatty acid metabolism. In addition, we observed that yor180cDelta cells fail to efficiently import the enzyme Delta3,Delta2-enoyl-CoA isomerase (Eci1p) to peroxisomes. This result suggested that Yor180Cp might associate with Eci1p in vivo, and a Yor180Cp-Eci1p interaction was detected using the yeast two-hybrid system. Potential roles for Yor180Cp in peroxisomal fatty acid metabolism are discussed. PMID- 10381340 TI - The incipient stage in thrombin-induced fibrin polymerization detected by FCS at the single molecule level. AB - We used fluorescence correlation spectroscopy (FCS) to study the activation of fibrinogen by thrombin and the subsequent aggregation of fibrin monomers into fibrin polymers at a very low and at physiological fibrinogen concentrations. In the labeling procedure used the fibrinogen was randomly labeled and the label was bound to the fibrinopeptide A and/or to the part of fibrinogen which after activation takes part in fibrin formation. We measured a diffusion coefficient for fibrinogen of 2.48 x 10(-7) +/- 0.10 x 10(-7) cm2/s. After activation with thrombin both fibrinopeptide A and fibrin polymerization products could be demonstrated. From our findings we suggest a model for the formation of a three dimensional network as two parallel processes, elongation and branching and that fibrin oligomers are not only intermediates in the polymerization process but also are substrates for branching. PMID- 10381341 TI - Antibodies against the PH domain of phospholipase C-delta1 inhibit Ins(1,4,5)P3 mediated Ca2+ release from the endoplasmic reticulum. AB - The pleckstrin homology domain (PH domain) is now well known as a structural module for the binding of inositol compounds. In the present study, polyclonal antibodies against the peptide KVKSSSWRRERFYK, derived from the N-terminal of the PH domain of phospholipase C-delta1 (PLC-delta1), were raised in rabbits. These were then tested for their ability to inhibit the binding of inositol 1,4,5 trisphosphate [Ins(1,4,5)P3] to the binding proteins including the receptor molecule. The Fab fragment of the antibodies but not the whole molecule inhibited the binding of Ins(1,4,5)P3 not only to PLC-delta1 but also to the Ins(1,4,5)P3 receptor, indicating that the antibodies raised recognized the binding site for Ins(1,4, 5)P3 in the receptor. Rat basophilic leukemic cells were permeabilized with saponin and assayed for Ins(1,4,5)P3-mediated Ca2+ release. Pretreatment of permeabilized RBL cells with the Fab fragment of the antibodies diminished the release of Ca2+ caused by Ins(1,4,5)P3, and further absorption experiments using a variety of synthetic peptides suggested that the tripeptide KVK is the epitope of the antibodies. Structural information about KVK will help in screening for Ins(1,4,5)P3 antagonists. PMID- 10381342 TI - Functional dissociation of anoikis-like cell death and activity of stress activated protein kinase. AB - Adhesion to the extracellular matrix is a crucial survival signal for epithelial and endothelial cells. Both cell types activate an endogenous death program termed "anoikis" when detached from the solid substratum. The signaling events culminating in anoikis are still unclear; recent studies have implicated Stress Activated Protein Kinase (SAPK), also known as Jun-N-Terminal kinase, as a potentially crucial signal transducer and mediator of anoikis. However, the generality and the causal role of SAPK in anoikis remain unclear and controversial. For these reasons we decided to examine the relationship between induction of anoikis and SAPK activation in three independent cell systems. We report here that in immortalized rat intestinal epithelial cells (IEC-18) and human umbilical vein endothelial cells (HUVEC), SAPK is activated weakly and transiently upon cell detachment while in canine kidney epithelial cells (MDCK) such induction is strong and protracted. However, cell types fail to commit to anoikis after remaining in three-dimensional culture for the time required for complete activation of SAPK. This suggests that there is no temporal correlation between SAPK activation and the onset of anoikis in any of the cell lines studied. We further examined the potential involvement of SAPK in the IEC-18 system by investigating a ras oncogene-transformed variant of IEC-18 cells (IEC 18 Ras 3) which are highly resistant to anoikis. Ras expression did not abrogate activation of SAPK, although these cells do exhibit altered kinetics of SAPK induction upon cell detachment. These results suggest that SAPK is not involved in anoikis regulation and that SAPK activation is likely a cell-type-specific epi phenomenon. PMID- 10381343 TI - Isolation and functional characterization of the 5'-upstream region of mouse P/Q type Ca2+ channel alpha1A subunit gene. AB - The omega-agatoxin-IVA-sensitive P/Q-type Ca2+ channel is predominantly expressed in the nervous system. To dissect the molecular mechanisms underlying the neuron specific expression of the P/Q-type channel, we have isolated and characterized the 5'-upstream region of the mouse alpha1A subunit gene. A transcription start site appeared to exist at -269 bp upstream from the start codon as found by 5' RACE analysis. The proximal promoter of the alpha1A subunit gene lacks a typical TATA box, but contains several transcription factor binding sequences, including two Sp1 sites. When linked to a placental alkaline phosphatase (PLAP) reporter gene to examine the promoter activity, the 6.3-kb (-6,273 to +269) 5'-upstream region, but not a smaller 3.0-kb construct (-3, 021 to +269), was able to drive the reporter gene in neuron-like PC12 cells. In contrast, neither of these constructs enhanced the PLAP expression in fibroblast NIH3T3 cells. The sequence between 6.3 and 3.0 kb of the 5'-upstream region did not show promoter activity in either of the cell lines, but enhanced TK promoter activity in PC12 cells, though not in NIH3T3 cells. These results suggest that neuron-specific elements of the alpha1A subunit gene are likely to be located in the distal upstream regions (-6,273 to -3,021) of the 5'-upstream sequence. PMID- 10381344 TI - The formation of oxidatively induced high-molecular-weight aggregate of alpha /gamma-crystallins. AB - alpha-/gamma-Crystallin interactions under oxidation with ascorbate-FeCl3-EDTA H2O2 followed by dialysis have been studied. A high-molecular-weight aggregate (HMWA) composed of alpha- and gamma-crystallin was observed for the mixture of the dialyzed alpha-crystallin and the oxidized gamma-crystallin through gel filtration chromatography. This illustrates an interaction between alpha crystallin and partially denatured gamma-crystallin induced by oxidation. No HMWA formation was observed under the condition without dialysis and/or with the addition of catalase to the oxidized gamma-crystallin prior to the addition of alpha-crystallin. More HMWA was formed by oxidized gamma-crystallin followed by the addition of alpha-crystallin than by simultaneous oxidation of both alpha- and gamma-crystallins. Conformational changes of alpha-crystallin during oxidation analyzed by circular dichroism spectra showed that oxidized alpha crystallin can gradually be restored to an ordered structure through dialysis. The overall results imply that structural changes of both alpha- and gamma crystallins and dialysis are required to form HMWA. The observation of this oxidatively induced chaperone/substrate complex suggests that an efficient chaperone-like protective action against oxidative insults may exist in vivo. PMID- 10381345 TI - Hox gene complexity in medaka fish may Be similar to that in pufferfish rather than zebrafish. AB - Changes in number and the genomic organization of Hox genes have played an important role in metazoan body-plan evolution. They make cluster(s), and in vertebrates, each cluster contains different number of Hox genes that have been classified into 13 groups. There are 39 Hox genes in four clusters on different chromosomes in the mammalian genome. In the fish, while 31 Hox genes in four clusters have been identified in pufferfish Fugu rubripes, 47 Hox genes in seven clusters exist in the zebrafish Danio rerio. To estimate the evolutionary origin of Hox organization in ray-finned fishes, we searched for Hox genes in the medaka fish Oryzias latipes, with a taxon thought to be widely separated from those of pufferfish and zebrafish. We synthesized various mixed oligonucleotides that can work as group-specific primers for PCR, then cloned and sequenced amplified fragments. Numbers of Hox genes identified in the present study were 2 for group 1, 2 for group 2, 1 for group 3, 3 for group 4, 6 for groups 5-7, 2 for group 8, 4 for group 9, 3 for group 10, 1 for group 12, and 3 for group 13. The primers specific for group 11 did not function in this study. Thus, at least 27 Hox genes are present in medaka genome, suggesting that the Hox gene complexity of the medaka genome is similar to that of the pufferfish rather than the zebrafish. PMID- 10381346 TI - Modulatory effect of HCO3- on rat mast cell exocytosis: cross-talks between bicarbonate and calcium. AB - HCO-3 modulation of histamine release and its relationship with the Ca2+ signal were studied in serosal rat mast cells. Histamine release was induced by Ca2+ mobilizing stimuli, namely compound 48/80, thapsigargin, Ca2+ chelators, ionophore A23187, and PMA and ionophore A23187 in a HCO-3-buffered medium or a HCO-3-free medium. The presence of HCO-3 reduced histamine release by 48/80, Ca2+ chelators, A23187, and PMA/A23187, but increased histamine release induced by thapsigargin. Histamine release by PMA was significantly higher in a HCO-3-free medium than in a HCO-3-free medium, as it was the PMA potentiation of histamine release by A23187. [Ca2+]i changes induced by these drugs were measured in fura-2 loaded mast cells. In thapsigargin and EGTA or BAPTA preincubated mast cells [Ca2+]i increase was higher in a HCO-3-buffered medium than in a HCO-3-free medium in the presence of Ca2+. On the contrary, in compound 48/80 and PMA/A23187 activated mast cells the [Ca2+]i increase is the same both in the presence and in the absence of HCO-3. The effect of HCO-3 on histamine release in serosal rat mast cells depends on the stimulus, but it is not related to the presence of Cl-. In thapsigargin-stimulated mast cells the effect of HCO-3 on histamine release may be related to the Ca2+ signal, but in compound 48/80, EGTA, and PMA/A23187 activated mast cells there is no relationship between intracellular Ca2+ and the inhibitory effect of HCO-3 on histamine release. Additionally, the PKC pathway is implicated in the inhibitory effect of HCO-3 on histamine release, the higher the chelation of calcium rendering the higher the enhancement of the response after adding calcium in the absence of HCO-3. PMID- 10381348 TI - The Glu residue in the conserved Asn-Glu-Pro sequence of endoglycoceramidase is essential for enzymatic activity. AB - Endoglycoceramidase (EGCase) is an enzyme capable of cleaving the glycosidic linkage between oligosaccharides and ceramides of various glycosphingolipids. We previously cloned the gene encoding EGCase II of Rhodococcus sp. M-777 and reported that the deduced amino acid sequence contained the Asn-Glu-Pro (NEP) sequence, conserved as part of the active site of family A cellulases (endo-1,4 beta-glucanases) (J. Biol. Chem. 272, 19846, 1997). The NEP sequence was also found in the deduced amino acid sequence of the newly cloned EGCase gene of Rhodococcus sp. C9. Replacement of the Glu residue in the NEP sequence with Gln or Asp by site-directed mutagenesis caused marked loss of enzymatic activity in both the M-777 and C9 EGCases but did not affect the expression of EGCase protein. This result clearly indicated that the NEP sequence is part of the active site of EGCase, in which the Glu residue plays an important role in the catalytic reaction, possibly in the same manner as in endo-1,4-beta-glucanase. PMID- 10381347 TI - Bistratene A induces a microtubule-dependent block in cytokinesis and altered stathmin expression in HL60 cells. AB - Bistratene A is a cyclic polyether which affects cell cycle progression and can induce phosphorylation of cellular proteins. Treatment of HL60 cells with 100 ng/ml bistratene A was found to inhibit cytokinesis but had no effect on DNA synthesis and nuclear division. Consequently, bistratene A-treated cells became polyploid and multinucleate. In association with the development of this phenotype, the cytoplasmic protein stathmin was biphasically phosphorylated and levels of expression were doubled. Immunostaining of binucleate cells (bistratene A for 24 h) revealed increased alpha-tubulin localization where the cleavage furrow might be expected to form, i.e., along the equatorial plane. Treatment of these binucleate cells with the microtubule depolymerizing agent nocadazole promoted cleavage furrow formation and partially ameliorated the bistratene A induced block in cell division. These findings implicate the polymerization status of microtubules and stathmin function in the regulation of cytokinesis. PMID- 10381350 TI - Secretory phospholipase A2 induces phospholipase Cgamma-1 activation and Ca2+ mobilization in the human astrocytoma cell line 1321N1 by a mechanism independent of its catalytic activity. AB - The effect of secretory phospholipase A2 (sPLA2) on intracellular Ca2+ signaling in human astrocytoma cells was studied. sPLA2 increased cytosolic [Ca2+] ([Ca2+]c) in both Ca2+-containing and Ca2+-free medium, thus suggesting Ca2+ release from intracellular stores. The activation by sPLA2 of arachidonate release via cytosolic PLA2 (cPLA2) was also independent of extracellular Ca2+. As sPLA2 requires Ca2+ for activity, these results indicate that both Ca2+ mobilization and cPLA2 activation induced by sPLA2 are unrelated to phospholipase activity but dependent on signaling mechanisms. The sPLA2-induced [Ca2+]c peak was sensitive to Bordetella pertussis toxin and inhibited by caffeine, suggesting its mediation by inositol 1,4,5-trisphosphate (IP3). sPLA2 induced tyrosine phosphorylation and membrane targeting of phospholipase Cgamma-1 (PLCgamma-1). Moreover, the Ca2+ peak was sensitive to protein tyrosine kinase inhibitors. sPLA2 activates two signaling pathways: one leading to the activation of the MAP kinase/cPLA2 cascade and another leading to PLCgamma activation and Ca2+ release. PMID- 10381349 TI - Impaired galactosylation of core 2 O-glycans in erythrocytes of beta1,4 galactosyltransferase knockout mice. AB - O- and N-glycans included in erythrocyte membrane glycoproteins from beta1,4 galactosyltransferase I (GalT-I) knockout mice were analyzed to examine how this enzyme deficiency affects glycosylation of proteins in erythroid cells. The results indicated that greater than 80% of core 2 O-glycans from GalT-I-/- mice are not galactosylated by beta1,4 linkage, resulting in the expression of Neu5Acalpha2 --> 3Galbeta1 --> 3(GlcNAcbeta1 --> 6)GalNAc, while core 2 O-glycans from GalT-I+/+ mice are fully galactosylated and occur as Neu5Acalpha2 --> 3Galbeta1 --> 3(Neu5Acalpha2 --> 3Galbeta1 --> 4GlcNAcbeta1 --> 6)GalNAc. On the other hand, beta1, 4-galactosylation of N-glycans of the mutant was approximately 60% that of the wild type. Thus, it is suggested that GalT-I is predominantly responsible for beta1,4-galactosylation of the core 2 O-glycan branch in erythroid cells. PMID- 10381351 TI - Induction of cell proliferation and apoptosis: dependence on the dose of the inducer. AB - Protein A (PA) of Staphylococcus aureus is known as an immunomodulator. In a search of the molecular mechanism(s) of PA-induced immunocyte potentiation, we found dose-dependent binding of PA (0.01 to 100 microg/ml PA) to the mice splenic lymphocytes. Interestingly, treatment of 1 microg PA/20 g mice increased the splenic lymphocyte number approximately 5-fold over control but at a 10-microg dose the cell number was decreased compared with a 1-microg dose. Flow cytometric analysis of cell-cycle phase distribution of nuclear DNA in splenic lymphocytes showed that at a 1-microg dose, PA shifted the cell-cycle phases from G0/G1 to S and G2/M supporting the pro-proliferative role of PA. In contrast, the same inducer increased the sub-G1 cell population at a 10-microg dose indicating the breakdown of cellular DNA. These findings were supported by DNA ladder formation and nuclear breakdown at this higher dose. Further studies revealed that at a 1 microg dose, the level of the pro-proliferative/anti-apoptotic protein bcl-2 was increased in splenic lymphocytes whereas at a 10-microg dose it showed a decreasing trend. In contrast, concentrations of proapoptotic proteins, p53 and bax, were increased at a 10-microg dose. A search of the mechanism(s) of such differential action of PA at these two doses revealed that the lower dose of PA upregulated the production of interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) to the extent which has already been reported by our laboratory to be beneficial to the host. However, at a larger dose, much higher release of TNF-alpha and interleukin-2 (IL-2) may account for the apoptosis of splenic cells. All these findings indicated that the cross-talk between all these pro- and anti-apoptotic factors may contribute to maintain a balance between growth and death of cells and may be one of the important factors deciding whether a cell would follow a proliferative pathway or an apoptotic pathway. PMID- 10381352 TI - Functional mimicry of protein A of Staphylococcus aureus by a proteolytically cleaved fragment. AB - Protein A (PA) of Staphylococcus aureus has an array of biological functions, such as antitumor, antitoxic, anticarcinogenic, immunomodulatory, antifungal, and antiparasitic properties. We have already established that a theoretical trypsin digested peptide fragment of protein A (20-mer) mimics immunomodulatory and IgG binding property of PA. In the present report we have concentrated on a 16-mer chymotryptic fragment of protein A, which has a sequence of 13 amino acids in common with the previously studied 20-mer peptide. Molecular modeling study qualitatively predicted that both 20-mer and 16-mer peptides retain Fc binding ability from an interaction energy point of view. In the present study our aim was to understand whether this theoretically predicted 16-mer chymotryptic fragment could be formed in a real experiment and also to understand its biological activities. Chymotrypsin cleavage of PA at 37 degrees C for 24 h produced four major fragments on reverse-phase HPLC. The amino acid analyses of each fragment show the absence of cysteine residue from all fragments, which justifies the absence of cysteine in PA. We also observed high content of aspartic acid and glutamic acid residues in all fragments. On gel-filtration chromatography the chymotrypsin cleavage of PA shows five peaks, one of which overlaps with our theoretically selected 16-mer peptide on superimposition. We verified the IgG binding capacity of 16-mer peptide by capillary electrophoresis. The 16-mer peptide also induces the production of TNFalpha and IL-1alpha in serum of mice. The above observations suggest that the 16-mer peptide may be produced by chymotrypsin cleavage and also that this peptide possesses some of the major biological properties of PA, such as IgG binding, TNFalpha and IL-1alpha elicitation, etc. PMID- 10381353 TI - Effects of antisense oligonucleotides to the cardiac Na+/Ca2+ exchanger on calcium dynamics in cultured cardiac myocytes. AB - The present study was designed to explore the role of the Na+/Ca2+ exchanger on spontaneous beating of cultured cardiac myocytes. Antisense oligonucleotides (AS) based on the sequence of the cardiac Na+/Ca2+ exchanger were used to decrease expression of this Ca2+ transporting protein in cardiac myocytes. An application of AS (10 microM) caused an increase in beating rate of myocytes within 6-24 h. After 24 h of exposure, AS increased the beating rate from an average rate of 77 beats/min in control and sense-treated myocytes to 103 beats/min. Moreover, myocytes treated for 24 h with 10 microM AS exhibited an increase in diastolic [Ca2+]i levels. The antisense treatment also led to a approximately 20% decrease in expression of Na+/Ca2+ exchanger proteins within 6-24 h. Changes in mRNA levels following AS treatment could not be detected within 3- to 24-h periods. The results of these studies suggest that the Na+/Ca2+ exchanger plays a potentiating role in spontaneous the beating process by regulating [Ca2+]i dynamics and that even a small reduction in the levels of the exchanger protein has marked effects on the handling of [Ca2+]i during the cardiac cycle. PMID- 10381355 TI - Alpha-SNAP functions in insulin exocytosis from mature, but not immature secretory granules in pancreatic beta cells. AB - To explore alpha-SNAP function in insulin exocytosis from either immature or mature secretory granules in pancreatic beta cells, we studied the effects of overexpression of adenovirus-mediated wild-type alpha-SNAP and C-terminally deleted alpha-SNAP mutant (1-285) on newly synthesized proinsulin and insulin release by rat islets and MIN6 cells. Rat islets overexpressing alpha-SNAP and mutant alpha-SNAP were pulse-chased. Exocytosis from immature and mature insulin secretory granules was measured as fractional (%) labeled-proinsulin release immediately after the pulse-labeling and percentage labeled-insulin release after a 3-h chase period, respectively. There was no difference in percentage labeled proinsulin release between the control and alpha-SNAP or mutant alpha-SNAP overexpressed islets. Although percentage labeled-insulin release after a 3-h chase period was significantly increased in alpha-SNAP-overexpressed islets, it was decreased in mutant alpha-SNAP-overexpressed islets. Thus, the results demonstrated that alpha-SNAP overexpression in rat islets primarily increased exocytosis from mature, but not immature insulin secretory granules. On the other hand, in MIN6 cells, alpha-SNAP overexpression scarcely affected glucose stimulated insulin release; therefore, we examined the effect of mutant alpha SNAP overexpression as the dominant-negative inhibitor on the newly synthesized proinsulin/insulin release using the same protocol as in the rat islet experiments. alpha-SNAP mutant (1-285) overexpression in MIN6 cells decreased the percentage labeled insulin release from mature secretory granules, but not percentage labeled proinsulin release from immature secretory granules. Thus, our data demonstrate that alpha-SNAP functions mainly in the mature insulin secretory granules in pancreatic beta cells. PMID- 10381354 TI - Association of bone mineral density with a polymorphism of the peroxisome proliferator-activated receptor gamma gene: PPARgamma expression in osteoblasts. AB - The peroxisome proliferator-activated receptor gamma (PPARgamma) protein as well as its transcript was detected in primary osteoblasts derived from rat calvariae. To analyze the possible involvement of PPARgamma in the human bone metabolism, association between bone mineral density (BMD) and a polymorphism of PPARgamma gene was investigated in Japanese postmenopausal women. We examined a polymorphism corresponding to a silent C --> T transition located in exon 6 of the PPARgamma gene, that was previously reported to be associated with plasma leptin levels in the obese. The frequencies of the C and T alleles in the population studied here were 0.851 and 0.149, respectively. When we separated the subjects into two groups, one bearing at least one T allele (CT + TT) and the other which did not (CC), the former subjects had lower BMD (Z score of total body; 0.056 +/- 1.00. L2-4; -0.25 +/- 1.26, mean +/- standard deviation). These data suggest that there is an association between the restriction fragment length polymorphism (RFLP) of PPARgamma gene and BMD and the possible involvement of this single nucleotide polymorphism (SNP) in the cause of postmenopausal osteoporosis in Japanese women. PMID- 10381356 TI - Abnormal dentatorubral-pallidoluysian atrophy (DRPLA) protein complex is pathologically ubiquitinated in DRPLA brains. AB - Dentatorubral-pallidoluysian atrophy (DRPLA) is caused by expansion of a glutamine repeat in DRPLA protein. DRPLA protein undergoes greater complex formation in DRPLA brain tissue, and expanded glutamine repeat enhances complex formation of DRPLA protein. Immunoblots with and without reduction show that the DRPLA protein complex is ubiquitinated only in DRPLA brain tissue. Moreover, immunoblots of regional DRPLA brain tissues reveal that pathological ubiquitination of DRPLA protein complex is found selectively in affected lesions. Double-labeling immunohistochemical studies with antibodies against DRPLA protein and ubiquitin demonstrate that the DRPLA protein is co-localized with ubiquitin in DRPLA neurons and show characteristic neuronal cytoplasmic inclusions with ubiquitinated DRPLA protein complex in the center. Our findings suggest that DRPLA protein undergoes abnormal complex formation with expanded glutamine repeat, and then the complex is pathologically ubiquitinated in DRPLA brain tissue. Pathological ubiquitination of abnormal DRPLA protein complex plays a role in DRPLA pathology. PMID- 10381357 TI - Activation of caspase-3 apoptotic pathways in skeletal muscle fibers in laminin alpha2-deficient mice. AB - dy/dy mice, which carry an unidentified mutation in the Lama2 gene, show dystrophic pathologies similar to those of human congenital muscular dystrophy. Laminin alpha2 deficiency induces apoptosis with DNA fragmentation. Caspases, which are involved in various types of cell death, are sequentially activated through a processing by other members of caspases. By using a cleavage site directed antibody against caspase-3 that specifically reacts with the active form of caspase-3, we immunochemically demonstrated that caspase-3 is activated in the skeletal muscle fiber of dy/dy mice and that some of the activated caspase-3 muscle fibers are TUNEL-positive. Thus the lack of laminin alpha2 signals activates caspase-3, resulting in the apoptosis of muscle fibers. PMID- 10381358 TI - Constitutive hsp70 is essential to mitosis during early cleavage of Paracentrotus lividus embryos: the blockage of constitutive hsp70 impairs mitosis. AB - Localization of constitutive hsp70 in eggs and early embryos of sea urchin Paracentrotus lividus is shown by means of in situ immunostaining. An accumulation of this protein is shown in the mitotic structures (asters, spindles and centrosomes). Microinjection of anti-hsp70 antibodies into eggs causes impairment of formation of mitotic structures and of cell division. This impairment goes from a complete mitotic block, to irregular mitotic apparatus formation with irregular cleavage, depending upon the antibody concentration. The localization of hsp70 after antibody microinjection is also described. Blockage of mitotic apparatus formation by nocodazole also blocks the concentration of hsp70 molecules observed in nontreated eggs. That the constitutive hsp70 plays a role in sea urchin mitosis is indicated. PMID- 10381359 TI - The length of polyglutamine tract, its level of expression, the rate of degradation, and the transglutaminase activity influence the formation of intracellular aggregates. AB - A common feature of CAG-expansion neurodegenerative diseases is the presence of intranuclear aggregates in neuronal cells. We have used a synthetic fusion protein containing at the NH2 terminus the influenza hemoagglutinin epitope (HA), a polyglutamine stretch (polyQ) of various size (17, 36, 43 CAG) and a COOH tail encoding the green fluorescent protein (GFP). The fusion proteins were expressed in COS-7 and neuroblastoma SK-N-BE cells. We found that the formation of aggregates largely depends on the length of polyglutamine tracts and on the levels of expression of the fusion protein. Moreover, transglutaminase overexpression caused an increase of insoluble aggregates only in cells expressing the mutant expanded protein. Conversely, treatment of cells with cystamine, a transglutaminase inhibitor, reduced the percentage of aggregates. We found also that the inhibition of the proteasome ubiquitin-dependent degradation increased the formation of intranuclear aggregates. These data suggest that length of polyglutamine tract, its expression, unbalance between cellular transglutaminase activity, and the ubiquitin-degradation pathway are key factors in the formation of intranuclear aggregates. PMID- 10381360 TI - TNFalpha-induced IEC-6 cell apoptosis requires activation of ICE caspases whereas complete inhibition of the caspase cascade leads to necrotic cell death. AB - Tumor necrosis factor (TNF)alpha is considered to play a key pathogenetic role in inflammatory bowel diseases. In this study we analyzed the mechanisms by which TNFalpha induces intestinal epithelial cell apoptosis. TNFalpha alone, and more potently in combination with IFNgamma, induced a high degree of IEC-6 cell apoptosis. This effect was more than 100-fold stronger if both of the TNF-R were stimulated, compared to stimulation of the p55-TNF-R alone, indicating an important apoptosis enhancing effect of the p75-TNF-R. TNFalpha-induced apoptosis required activation of ICE caspases and was completely abolished by its inhibitor, zVAD-fmk. Specific inhibition of caspase-3 with zDEVD-fmk did not alter the effect of TNFalpha. Western blot analyses confirmed that caspase-3 was not activated in response to TNFalpha. In the presence of complete inhibition of the caspase cascade with zVAD-fmk (>/=50 microM), TNFalpha induced cell necrosis rather than apoptosis. Our data reveal that TNFalpha can trigger enterocyte cell death via apoptosis or necrosis, depending upon the activation or blockade of specific caspases. PMID- 10381361 TI - Fas/Fas ligand system in prolactin-induced apoptosis in rat corpus luteum: possible role of luteal immune cells. AB - The prolactin (PRL) surge in cycling rats during the proestrous afternoon is an inducer of apoptotic cell death in luteal cells. This luteolytic action of PRL is peculiar, because PRL may be categorized as a survival factor, if other known physiological functions of PRL are taken into account. Here we analyzed the underlying molecular/cellular mechanisms of this PRL-induced apoptosis. Corpora lutea (CL) were prepared from the ovary on the proestrous day and cultured with or without PRL (2 microg/ml). An addition of PRL to the culture medium induced DNA breakdown in the nuclei of cells mostly identified as steroidogenic by 3beta HSD activity staining, and the number of 3beta-HSD-positive cells were significantly decreased, indicating the induction of apoptotic cell death by PRL among luteal cells in culture. Next, the expression of membrane form-Fas ligand (mFasL) in the luteal cell lysate was quantified, because Fas receptor is known to have an exact physiological role in luteolysis. An addition of PRL increased the expression of mFasL. Immunostaining and TUNEL assay on regressing CL revealed that both CD3-positive cells and FasL-positive cells were co-localized in the regions where apoptosis convergently occurred. Moreover, an addition of concanavalin A (ConA), a T-cell specific activator, to the culture mimicked the PRL action by inducing apoptosis in luteal cells and enhancing the expression of mFasL. These data suggest that the CD3-positive T lymphocyte in the CL is at least one of the PRL-effector cell species during the process of luteolysis in rats, and that FasL expression of these cells is upregulated by PRL. PMID- 10381362 TI - Cloning and characterization of a novel orphan G-protein-coupled receptor localized to human chromosome 2p16. AB - We report the identification and characterisation of a novel human orphan G protein-coupled receptor (GPR) which maps to chromosome 2p16. We have determined the full-length coding sequence and genomic structure of a gene corresponding to the anonymous expressed sequenced tag, WI-31133. This gene encodes a novel protein that is 540 amino acids in length. Protein sequence analysis predicts the presence of seven transmembrane domains, a characteristic feature of GPRs. In situ hybridisation to human retina and Northern blot analysis of human retinal pigment epithelium (RPE) showed localisation of this transcript to the RPE and cells surrounding retinal arterioles. In contrast, the transcript was localised to the photoreceptor inner segments and the outer plexiform layer in mouse sections. Northern blot analysis demonstrated a 7 kb transcript highly expressed in the brain. No mutations were identified during a screen of patients suffering from Doyne's honeycomb retinal dystrophy (DHRD), an inherited retinal degeneration which maps to chromosome 2p16. PMID- 10381363 TI - cis-2,3-dihydro-2,3-dihydroxybiphenyl dehydrogenase and cis-1, 2-dihydro-1,2 dihydroxynaphathalene dehydrogenase catalyze dehydrogenation of the same range of substrates. AB - Pseudomonas putida strain G7 cis-1,2-dihydro-1, 2-dihydroxynaphthalene dehydrogenase (NahB) and Comamonas testosteroni strain B-356 cis-2,3-dihydro-2,3 dihydroxybiphenyl dehydrogenase (BphB) were found to be catalytically active towards cis-2,3-dihydro-2,3-dihydroxybiphenyl (specificity factors of 501 and 5850 s-1 mM-1 respectively), cis-1,2-dihydro-1, 2-dihydroxynaphthalene (specificity factors of 204 and 193 s-1 mM-1 respectively) and 3,4-dihydro-3,4 dihydroxy-2,2',5, 5'-tetrachlorobiphenyl (specificity factors of 1.6 and 4.9 s-1 mM-1 respectively). A key finding in this work is the capacity of strain B-356 BphB as well as Burkholderia cepacia strain LB400 BphB to catalyze dehydrogenation of 3,4-dihydro-3,4-dihydroxy-2,2',5, 5'-tetrachlorobiphenyl which is the metabolite resulting from the catalytic meta-para hydroxylation of 2,2',5,5'-tetrachlorobiphenyl by LB400 biphenyl dioxygenase. PMID- 10381364 TI - Substitution of the two carboxyl-terminal serines by alanine causes retention of MAL, a component of the apical sorting machinery, in the endoplasmic reticulum. AB - MAL, a selective resident of glycolipid-enriched membranes (GEMs), is an integral membrane protein necessary for apical transport and accurate sorting of the influenza virus hemagglutinin in MDCK cells. The carboxyl-terminal end of MAL has the sequence Phe-Ser-Leu-Ile-Arg-Trp-Lys-Ser-Ser (FSLIRWKSS), which includes the LIRW motif necessary for sorting MAL to GEMs, and whose last five amino acids resemble dilysine-based signals involved in endoplasmic reticulum (ER) retention. We have addressed the influence of the carboxyl-terminal serines in both MAL distribution and incorporation into GEMs. Substitution of the serines by alanine impeded the access of MAL to GEMs and changed its distribution from a perinuclear distribution to an ER pattern. The RWKSS sequence appended to the carboxyl terminus of CD4 caused retention of the chimera in the ER. Thus, although this pentapeptide can function producing ER retention in other protein context, the presence of the carboxyl-terminal serines in the intact MAL molecule prevents its use as an ER-retention signal. PMID- 10381365 TI - Phosphorylation of the cyclosome is required for its stimulation by Fizzy/cdc20. AB - Exit from mitosis in eukaryotic cells is regulated by the cyclosome (also called anaphase promoting complex or APC), a multisubunit ubiquitin ligase that acts on mitotic cyclins. Previous studies in a cell-free system from clam oocytes have shown that the activation of the cyclosome at the end of mitosis involves its phosphorylation by protein kinase Cdk1/cyclin B. Genetic and biochemical studies have furthermore indicated that cyclosome activity also requires a WD-40 repeat containing protein called Fizzy (FZY) or Cdc20. It has been suggested [Fang et al. (1998) Mol. Cell 2, 163-171] that in the presence of FZY, the phosphorylation of the cyclosome is not critical for its activation. By contrast, we find that the activity of the interphase, non-phosphorylated form of the cyclosome from clam embryos is not stimulated by FZY to a significant extent. However, when interphase cyclosome is first incubated with protein kinase Cdk1/cyclin B, the subsequent supplementation of FZY greatly stimulates its cyclin-ubiquitin ligase activity. Furthermore, phosphatase treatment of purified mitotic cyclosome prevents its stimulation by FZY, a process that can be reversed by the action of protein kinase Cdk1/cyclin B. We conclude that in the early embryonic cell cycles, the primary event in the activation of the cyclosome at the end of mitosis is its Cdk1-dependent phosphorylation and activation by FZY takes place in a subsequent process. PMID- 10381366 TI - Glucuronidation of 7-ethyl-10-hydroxycamptothecin (SN-38) by the human UDP glucuronosyltransferases encoded at the UGT1 locus. AB - 7-Ethyl-10-hydroxycamptothecin (SN-38) is a very promising anticancer drug used for the treatment of metastatic colonrectal cancer. SN-38 is the active metabolite of irinotecan, a semisynthetic anticancer drug derived from 20(S)camptothecin. In this study, we examined the potential for each of the UGT1 encoded isoforms (UGT1A1 and UGT1A3 through UGT1A10) to glucuronidate SN-38. The amount of specific protein for each isoform was determined by Western blot analysis. Although UGT1A1 was previously shown to metabolize this drug, the results of this study show that UGT1A7 glucuronidates this chemical at a 9- to 21 fold higher level at pH 6. 4 and pH 7.6, respectively, than that by UGT1A1. The activity of UGT1A7 is from 8.4- to 19-fold higher at pH 6.4 and 12- to 40-fold higher at pH 7.6 than that by the other 7 UGT1 encoded isoforms. UGT1A7 glucuronidates SN-38 with an apparent Km of 5 microM. Hence, the distribution of this isoform in the gastrointestinal tract has the potential to impact the effectiveness of this chemotherapeutic agent. PMID- 10381368 TI - Biochemical and spectroscopic characterization of overexpressed fuscoredoxin from Escherichia coli. AB - Fuscoredoxin is a unique iron containing protein of yet unknown function originally discovered in the sulfate reducers of the genus Desulfovibrio. It contains two iron-sulfur clusters: a cubane [4Fe-4S] and a mixed oxo- and sulfido bridged 4Fe cluster of unprecedented structure. The recent determination of the genomic sequence of Escherichia coli (E. coli) has revealed a homologue of fuscoredoxin in this facultative microbe. The presence of this gene in E. coli raises interesting questions regarding the function of fuscoredoxin and whether this gene represents a structural homologue of the better-characterized Desulfovibrio proteins. In order to explore the latter, an overexpression system for the E. coli fuscoredoxin gene was devised. The gene was cloned from genomic DNA by use of the polymerase chain reaction into the expression vector pT7-7 and overexpressed in E. coli BL21(DE3) cells. After two chromatographic steps a good yield of recombinant protein was obtained (approximately 4 mg of pure protein per liter of culture). The purified protein exhibits an optical spectrum characteristic of the homologue from D. desulfuricans, indicating that cofactor assembly was accomplished. Iron analysis indicated that the protein contains circa 8 iron atoms/molecule which were shown by EPR and Mossbauer spectroscopies to be present as two multinuclear clusters, albeit with slightly altered spectroscopic features. A comparison of the primary sequences of fuscoredoxins is presented and differences on cluster coordination modes are discussed on the light of the spectroscopic data. PMID- 10381367 TI - EDG3 is a functional receptor specific for sphingosine 1-phosphate and sphingosylphosphorylcholine with signaling characteristics distinct from EDG1 and AGR16. AB - AGR16/H218/EDG5 and EDG1 are functional receptors for lysosphingolipids, whereas EDG2 and EGD4 are receptors for lysophosphatidic acid (LPA). The present study demonstrates that EDG3, the yet poorly defined member of the EDG family G protein coupled receptors, shows identical agonist specificity, but distinct signaling characteristics, compared to AGR16 and EDG1. Overexpression of EDG3 conferred a specific [32P]S1P binding, which was displaced by S1P and sphingosylphosphorylcholine (SPC), but not by LPA or other related lipids. In cells overexpressing EDG3, S1P induced inositol phosphate production and [Ca2+]i increase in a manner only partially sensitive to pertussis toxin (PTX), which was similar to the case of AGR16, but quite different from the case of EDG1, in which the S1P-induced responses were totally abolished by PTX. EDG3 also mediated activation of mitogen-activated protein kinase (MAPK) in PTX-sensitive and Ras dependent manners, as in the cases of EDG1 and AGR16, although EDG3 and EDG1 were more effectively coupled to activation of MAPK, compared to AGR16. Additionally, EDG3 mediated a decrease in cellular cyclic AMP content, like EDG1, but contrasting with AGR16 which mediated an increase in cyclic AMP. These and previous results establish that EDG1, AGR16 and EDG3 comprise the lysosphingolipid receptor subfamily, each showing distinct signaling characteristics. PMID- 10381369 TI - Identification of cDNAs for Sox-4, an HMG-Box protein, and a novel human homolog of yeast splicing factor SSF-1 differentially regulated during apoptosis induced by prostaglandin A2/delta12-PGJ2 in Hep3B cells. AB - We have examined specific genes whose expression is altered during apoptosis induced by prostaglandin (PG)A2 and Delta12-PGJ2 in human hepatocellular carcinoma Hep3B cells. Using mRNA differential display, we have identified two genes: one is specifically up-regulated and encodes for human Sox-4 (Sry-HMG box gene) and the other is significantly down-regulated and is the human homolog of yeast Ssf-1, a novel splicing factor. Northern blot analysis confirmed their differential expressions. Interestingly, Sox-4 was highly expressed in subcutaneous tumors grown in nude mice as a xenograft from Hep3B cells. These results suggest that the expression of Sox-4 may be related to the apoptosis pathway leading to cell death as well as to tumorigenesis, and that Ssf-1 gene may serve as a negative regulator of PGA2/Delta12-PGJ2-mediated Hep3B cell apoptosis. PMID- 10381370 TI - Protease-sensitive urinary pheromones induce region-specific Fos-expression in rat accessory olfactory bulb. AB - Vomeronasal organs of female Wistar rats were exposed with sprayed urine preparations of male Wistar rats prior to sacrifice. Exposure to crude urine and ultrafiltrated urine preparation (<5000 Da) induced significant Fos expression, which is correlated with cellular activity, in the mitral/tufted cell layer of the accessory olfactory bulb (AOB), while exposure to the remaining substances after ultrafiltration (>5000 Da) and control salt solution did not. Exposure to urine preparation treated with papain induced expression of Fos-immunoreactive cells in the rostral region of the AOB, but did not induce such expression in the caudal region. Exposure to urine preparation treated with pronase induced urine specific Fos immunoreactivity neither in the rostral nor in the caudal region. These results suggest that at least two different peptides carrying pheromonal activities are contained in male Wistar rat urine. PMID- 10381371 TI - Hepatocyte growth Factor/Scatter factor (HGF/SF) is a regulator of fibronectin splicing in MDCK cells: comparison between the effects of HGF/SF and TGF-beta1 on fibronectin splicing at the EDA region. AB - EDA-containing fibronectin (EDA + FN) is selectively produced under several physiological and pathological conditions requiring tissue remodeling, where cells actively proliferate and migrate. Only a few growth factors, such as transforming growth factor (TGF)-beta1, have been reported to regulate FN splicing at the EDA region. In the present study, we showed for the first time that hepatocyte growth factor/scatter factor (HGF/SF), which is mainly produced by mesenchymal cells and functions as a motogenic and mitogenic factor for epithelial cells, modulates FN splicing at the EDA region in MDCK epithelial cells. HGF/SF treatment increased the ratio of EDA + FN mRNA to mRNA of FN that lacks EDA (EDA - FN) (EDA+/EDA- ratio) more than TGF-beta1 treatment did: at a range from 0.02 to 20 ng/ml, HGF/SF increased the ratio in a dose-dependent manner by up to 2. 1-fold compared with nontreated control, while TGF-beta1 stimulated the EDA+/EDA- ratio by 1.5-fold at the optimum dose of 10 ng/ml. However, TGF-beta1 increased total FN mRNA levels by 3-fold at 10 ng/ml, but HGF/SF did not. We previously demonstrated that fibroblasts cultured at low cell density expressed more EDA + FN than those at high cell density. The same effect of cell density was also observed in MDCK cells. Furthermore, at low cell density, HGF/SF stimulated EDA inclusion into FN mRNA more effectively than did TGF-beta1, whereas at high cell density, TGF-beta1 was more potent than HGF/SF. Simultaneous treatment of cells with HGF/SF and TGF-beta1 synergistically stimulated EDA inclusion into FN mRNA. This stimulation of EDA inclusion into FN mRNA by HGF/SF led to increased EDA + FN protein production and secretion by cells, which was demonstrated by immunoblotting. Thus, our studies have shown that HGF/SF is an enhancer of EDA inclusion into FN mRNA as is TGF-beta1. However, these two factors were different in their effects at low and high cell densities and also in their effects on total FN mRNA levels. PMID- 10381373 TI - Noggin and bone morphogenetic protein-4 coordinately regulate the progression of chondrogenic differentiation in mouse clonal EC cells, ATDC5. AB - Here we report the gene expression and regulation and the function of noggin in clonal mouse chondrogenic EC cells, ATDC5. In ATDC5 cells, the expression of Noggin mRNA increased in parallel with the progression of chondrogenic differentiation. The treatment with conditioned medium of noggin-transfected COS 7 cells decreased the levels of type II and type X collagen gene transcripts of differentiated ATDC5 cells in a dose-dependent manner, and this inhibitory action was reversed by exogenously administered BMP-4 in a dose-dependent manner. The steady-state level of noggin gene transcripts was markedly upregulated by exogenously administered BMP-4 in time- and dose-dependent manners. Furthermore, this stimulatory effect of BMP-4 was attenuated by treatment with actinomycin D, but not with cycloheximide. These results indicate that noggin and BMP-4 coordinately regulate the progression of chondrogenic differentiation in ATDC5 cells. PMID- 10381372 TI - Differentiation of U937 cells enables a phospholipase D-dependent pathway of cytosolic phospholipase A2 activation. AB - Treatment with dibutyryl cyclic AMP (dBcAMP) of the human, premonocytic U937 cell line results in differentiation toward a monocyte/granulocyte-like cell. This differentiation enables the cell to activate cytosolic phospholipase A2 (cPLA2) to release arachidonate upon stimulation. In contrast, undifferentiated cells are unable to release arachidonate even when stimulated with calcium ionophores. In the present research, a role for phospholipase D (PLD) in the regulation of cPLA2 was shown based on a number of observations. First, the ionomycin- and fMLP stimulated production of arachidonate in differentiated cells was sensitive to ethanol (2% (v/v)). Ethanol acts as an alternate substrate in place of water for PLD producing phosphatidylethanol (PEt) instead of phosphatidic acid. Indeed, ionomycin stimulation of differentiated cells produced a 14-fold increase in PEt levels. Further evidence for the involvement of PLD in the regulation of cPLA2 came from the observation that the stimulated production of diacylglycerol (for which phosphatidic acid is a major source) was greatly diminished in undifferentiated cells as compared to differentiated cells. Moreover, the normally deficient activation of cPLA2 in undifferentiated cells could be stimulated to release arachidonate if the cells were electroporated in the presence of GTP[gamma]S and MgATP. This treatment stimulates phosphatidylinositol 4,5-bisphosphate (PIP2) production which appears to activate PLD and cPLA2 in subsequent steps. The phosphatidic acid (and diacylglycerol derived from phosphatidic acid) appears to greatly regulate the action of cPLA2 by an unknown mechanism, and undifferentiated cells lack the ability to stimulate PLD activity due to a dysfunction of PIP2 production. PMID- 10381374 TI - Cloning, expression of the psbU gene, and functional studies of the recombinant 12-kDa protein of photosystem II from a red alga Cyanidium caldarium. AB - The encoding extrinsic 12-kDa protein of oxygen-evolving PS II complex from a red alga, Cyanidium caldarium, was cloned and sequenced by means of PCR and a rapid amplification of cDNA ends (RACE) procedure. The gene encodes a putative polypeptide of 154 amino acids with a calculated molecular mass of 16,714 Da. The full sequence of the protein includes two characteristic transit peptides, one for transfer across the chloroplast envelope and another for targeting into the thylakoid lumen. This indicates that the protein is encoded in the nuclear genome. The mature protein consists of 93 amino acids with a calculated molecular mass of 10,513 Da. The cloned gene was successfully expressed in Escherichia coli and the resulting protein was purified, reconstituted to CaCl2-washed PS II complex together with the other extrinsic proteins of 33 and 20 kDa and cyt c 550. The recombinant 12-kDa protein bound completely with the PSII complex, which resulted in a restoration of oxygen evolution equal to the level achieved by binding of the native 12-kDa protein. PMID- 10381375 TI - Denervation of chicken skeletal muscle causes an increase in acetylcholinesterase mRNA synthesis. AB - We have examined the changes in enzymatic activity and the levels of transcripts for AChE following denervation of chicken skeletal muscle. Quantitation of RNA blots indicates that AChE transcripts are increased following denervation. AChE transcripts increased approximately 17-fold in the fast-twitch posterior latissimus dorsi muscle and approximately 4-fold in the tonic anterior latissimus dorsi muscle 10 days after denervation of adult chickens. Both AChE transcript levels and enzyme activity increased in parallel for the two muscles. AChE transcripts also increased approximately 4-fold in the shank muscles of 2-day-old chicks following denervation. Transcript synthesis, measured by run-on transcription, increased approximately 3-fold in these denervated muscles. These results suggest that the increase in AChE transcripts following denervation in the chicken is due, at least in part, to an increase in the rate of its synthesis. PMID- 10381376 TI - Apoptosis and mitotic arrest are two independent effects of the protein phosphatases inhibitor okadaic acid in K562 leukemia cells. AB - Treatment of human myeloid leukemia K562 cells with the serine/threonine protein phosphatases inhibitor okadaic acid induced mitotic arrest followed by apoptosis in a synchronized manner. The effect was observed at drug concentrations that inhibited the protein phosphatase type 2A but not type 1. We investigated whether apoptosis was a consequence of the preceding mitosis arrest or was induced independently by okadaic acid. We found that (1) apoptosis, but not mitotic arrest, was inhibited in cells with constitutive expression of Bcl-2; (2) pretreatment of cells with the DNA synthesis inhibitor hydroxyurea blocked the mitotic arrest but not the apoptosis mediated by okadaic acid; (3) down regulation of c-myc gene was associated with apoptosis, but not with mitotic arrest; and (4) inhibition of protein synthesis abrogated mitotic arrest, but not apoptosis. The results suggest that inhibition of protein phosphatase 2A by okadaic acid provokes mitotic arrest and apoptosis of leukemia cells by independent mechanisms. PMID- 10381377 TI - Molecular cloning of rat SH2-containing inositol phosphatase 2 (SHIP2) and its role in the regulation of insulin signaling. AB - SH2-containing inositol 5'-phosphatase (SHIP) plays a negative regulatory role in hematopoietic cells. We have now cloned the rat SHIP isozyme (SHIP2) cDNA from skeletal muscle, which is one of the most important target tissue of insulin action. Rat SHIP2 cDNA encodes a 1183-amino-acid protein that is 45% identical with rat SHIP. Rat SHIP2 contains an amino-terminal SH2 domain, a central 5' phosphoinositol phosphatase activity domain, and a phosphotyrosine binding (PTB) consensus sequence and a proline-rich region at the carboxyl tail. Specific antibodies to SHIP2 were raised and the function of SHIP2 was studied by stably overexpressing rat SHIP2 in Rat1 fibroblasts expressing human insulin receptors (HIRc). Endogenous SHIP2 underwent insulin-mediated tyrosine phosphorylation and phosphorylation was markedly increased when SHIP2 was overexpressed. Although overexpression of SHIP2 did not affect insulin-induced tyrosine phosphorylation of the insulin receptor beta-subunit and Shc, subsequent association of Shc with Grb2 was inhibited, possibly by competition between the SH2 domains of SHIP2 and Grb2 for the Shc phosphotyrosine. As a result, insulin-stimulated MAP kinase activation was reduced in SHIP2-overexpressing cells. Insulin-induced tyrosine phosphorylation of IRS-1, IRS-1 association with the p85 subunit of PI3-kinase, and PI3-kinase activation were not affected by overexpression of SHIP2. Interestingly, although both PtdIns-(3,4,5)P3 and PtdIns(3,4)P2 have been implicated in the regulation of Akt activity in vitro, overexpression of SHIP2 inhibited insulin-induced Akt activation, presumably by its 5'-inositol phosphatase activity. Furthermore, insulin-induced thymidine incorporation was decreased by overexpression of SHIP2. These results indicate that SHIP2 plays a negative regulatory role in insulin-induced mitogenesis, and regulation of the Shc. Grb2 complex and of the downstream products of PI3-kinase provides possible mechanisms of SHIP2 action in insulin signaling. PMID- 10381378 TI - Characterization of five human cDNAs with homology to the yeast SIR2 gene: Sir2 like proteins (sirtuins) metabolize NAD and may have protein ADP ribosyltransferase activity. AB - The yeast Sir2 protein regulates epigenetic gene silencing and as a possible antiaging effect it suppresses recombination of rDNA. Studies involving cobB, a bacterial SIR2-like gene, have suggested it could encode a pyridine nucleotide transferase. Here five human sirtuin cDNAs are characterized. The SIRT1 sequence has the closest homology to the S. cerevisiae Sir2p. The SIRT4 and SIRT5 sirtuins more closely resemble prokaryotic sirtuin sequences. The five human sirtuins are widely expressed in fetal and adult tissues. Recombinant E. coli cobT and cobB proteins each showed a weak NAD-dependent mono-ADP-ribosyltransferase activity using 5, 6-dimethylbenzimidazole as a substrate. Recombinant E. coli cobB and human SIRT2 sirtuin proteins were able to cause radioactivity to be transferred from [32P]NAD to bovine serum albumin (BSA). When a conserved histidine within the human SIRT2 sirtuin was converted to a tyrosine, the mutant recombinant protein was unable to transfer radioactivity from [32P]NAD to BSA. These results suggest that the sirtuins may function via mono-ADP-ribosylation of proteins. PMID- 10381379 TI - A new allele, DNASE1*6, of human deoxyribonuclease I polymorphism encodes an Arg to Cys substitution responsible for its instability. AB - A new allele, DNASE1*6, of human deoxyribonuclease I (DNase I) has been discovered by isoelectric focusing: its gene product has the most cathodic pI of the six electrophoretic variants. Results of DNA sequencing, mismatched PCR restriction fragment length polymorphism, and transient transfection of the variant construct showed that the mutant was caused by a C-T transition at nucleotide position 1826, resulting in an Arg to Cys substitution at amino acid position 185 of the mature enzyme. The variant isoenzyme, expressed in COS-7 cells, was more labile than the other types. Instability and an increase in the pI value of the variant suggest that a structural alteration, perhaps due to aberrant formation of a disulfide bond, could occur in the enzyme. PMID- 10381380 TI - Secondary structure formation is the earliest structural event in the refolding of an all beta-sheet protein. AB - The refolding kinetics of cobrotoxin (CBTX), a small-molecular-weight ( approximately 7 kDa) all beta-sheet protein, has been monitored using a variety of biophysical techniques. The secondary structure formation and hydrophobic collapse occur as distinct events during the refolding of the protein. Complete secondary structure formation occurs prior to the clustering of the hydrophobic residues. The late stage(s) of the refolding pathway of CBTX is characterized by change(s) in the local environment and optical asymmetry of the indole ring of the sole tryptophan residue. The results obtained in the present study, to our knowledge, represent the first unambiguous experimental support for the framework model of protein folding. PMID- 10381381 TI - MAP kinase-independent induction of proto-oncogene c-fos mRNA by hemin in human cells. AB - Treatment of HeLa cells or human skin fibroblast cells with hemin led to a time- and dose-dependent rapid induction of c-fos mRNA. This induction was absent in the cells treated with actinomycin D, indicating that the c-fos induction by hemin occurs at the level of transcription. Metalloporphyrins, including zinc-, cobalt-, and tin-protoporphyrin, ferric ion, and protoporphyrin also induced c fos mRNA. Transient reporter assay with the reporter constructs of the human c fos gene promoter up to -404 bp connected to the luciferase gene showed high activity but no induction by hemin, suggesting that cis-acting elements, including the serum response element located about -310 bp upstream of the human c-fos gene promoter, may not contribute to the heme-dependent induction. With in gel assay of protein kinases, the activity of the mitogen-activated protein (MAP) kinases such as extracellular signal-regulated kinase 12 or p38 MAP kinase in hemin-treated HeLa cells was not stimulated. Stimulation of c-Jun N-terminal kinase by hemin was nil. Furthermore, PD58059 and SB203580, inhibitors for MAP kinases, did not affect the hemin-dependent c-fos induction. Of the inhibitors for protein kinases so far tested, KN-62, a specific inhibitor for calmodulin dependent protein kinase II (CaMK II), inhibited the induction of c-fos mRNA by hemin. Phosphorylation of CaMK II in hemin-treated cells increased. With gel mobility assay, the DNA AP-1 binding activity transiently increased when treating HeLa cells with hemin. Therefore, induction of c-fos led to an activation of AP-1 in the presence of hemin. We suggest that calmodulin-dependent protein kinase II rather than the MAP kinase family regulates the induction of the human c-fos gene expression by hemin. PMID- 10381382 TI - Identification of two human WAVE/SCAR homologues as general actin regulatory molecules which associate with the Arp2/3 complex. AB - WAVE/SCAR protein was identified as a protein which has similarity to WASP and N WASP, especially in its C terminal. Recently, WAVE/SCAR protein has been shown to cooperate with the Arp2/3 complex, a nucleation core for actin polymerization in vitro. However, in spite of its general function, WAVE/SCAR expression is mainly restricted to the brain, suggesting the existence of related molecule(s). We here identified two human WAVE/SCAR homologues, which cover other organs. We named the original WAVE1 and newly identified ones WAVE2 and WAVE3. WAVE2 had a very wide distribution with strong expression in peripheral blood leukocytes and mapped on chromosome Xp11.21, next to the WASP locus. WAVE3 and WAVE1 had similar distributions. WAVE3 was strongly expressed in brain and mapped on chromosome 13q12. WAVE1 was mapped on chromosome 6q21-22. Ectopically expressed WAVE2 and WAVE3 induced actin filament clusters in a similar manner with WAVE1. These actin cluster formations were suppressed by deletion of their C-terminal VPH (verproline homology)/WH2 (WASP homology 2) domain. Further, WAVE2 and WAVE3 associate with the Arp2/3 complex as does WAVE1. Our identification of WAVE homologues suggests that WAVE family proteins have general function for regulating the actin cytoskeleton in many tissues. PMID- 10381383 TI - Plakophilin-3, a novel armadillo-like protein present in nuclei and desmosomes of epithelial cells. AB - We report on a novel Armadillo-like protein, termed plakophilin-3. The human protein, which is encoded by a 2.8 kb messenger RNA, has a predicted molecular mass of 87 kDa. The protein comprises 10 Armadillo-like repeats, preceded by an amino-terminal region of 293 amino acid residues and followed by a short carboxy terminal region of 27 amino acid residues. Plakophilin-3 is classified as a member of the p120(ctn)/plakophilin subfamily of Armadillo proteins based on the number and organization of the Armadillo repeats and its high sequence similarity to other members of this family. CLUSTAL W alignment of p120(ctn)/plakophilin subfamily members showed the plakophilin-3 protein to be most similar to plakophilin-1 and -2. Western blot analysis of plakophilin-3 revealed expression in all epithelial cell lines tested but not in foreskin fibroblasts and various sarcoma-derived cell lines. This is unlike most other members of the p120(ctn)/plakophilin subfamily, which are widely expressed. By immunofluorescence, the plakophilin-3 protein was colocalized with desmoglein in desmosomes of epithelial cells. In addition, an intriguing speckle-like nuclear staining was observed. Hence, like plakophilin-1 and -2, plakophilin-3 displays a dual intracellular location, i.e. in the desmosomal plaque and in the nucleus. These results suggest the involvement of plakophilin-3 in both desmosome dependent adhesion and signaling pathways. Furthermore, the human plakophilin-3 gene was mapped on the chromosomal locus 11p15 by fluorescent in situ hybridization. PMID- 10381384 TI - Contribution of plus and minus end pathways to microtubule turnover. AB - Turnover is important for the maintenance and remodeling of the cytoskeleton during the processes of cell morphogenesis, mitosis and motility. Microtubule (MT) turnover is thought to occur by dynamic instability, growth and shortening at distal (plus) ends. Recent observation of MT release from the centrosome and depolymerization from proximal (minus) ends indicates the existence of a minus end pathway. To evaluate the relative contributions of plus and minus end pathways to turnover, we analyzed MT dynamics in a model system, the fish melanophore, a large non-motile cell with a regular radial array of long MTs. MT ends were tracked in digital fluorescence time-lapse sequences and life histories of individual MTs were analyzed using random walk theory generalized to the case of diffusion with drift. Analysis of plus end dynamics gave an apparent diffusion coefficient of D=7.5 microm2/minute. The random walk model predicts that the half time for turnover driven solely by plus end dynamics will depend strongly on position in the cell. Based on the experimentally determined value of D, turnover of MTs near the center of a typical melanophore of radius 70 microm was calculated to require over 5 hours, a paradoxically long time. To examine MT behavior deep in the cytoplasm, we developed a novel, sequential subtraction mode of image analysis. This analysis revealed a subpopulation of MTs which shortened from their minus ends, presumably after constitutive release from the centrosome. Given the relative slowness of plus end dynamics to turn over the root of a long MT, the turnover of MTs near the cell center is determined primarily by the minus end pathway. MTs released from the centrosome become replaced by newly nucleated ones. The relative contributions of plus and minus end pathways was estimated from the diffusion coefficient, D, for the plus end, the length distribution of MTs, t he frequency of free minus ends, and the rate of minus-end shortening. We conclude that, in large animal cells with a centrosomally focussed array of MTs, turnover occurs by a combination of plus and minus end pathways, the plus end dominating at the cell periphery and the minus end dominating near the cell center. PMID- 10381385 TI - Intracellular traffic of the MHC class I-like IgG Fc receptor, FcRn, expressed in epithelial MDCK cells. AB - Transfer of passive immunity from mother to the fetus or newborn involves the transport of IgG across several epithelia. Depending on the species, IgG is transported prenatally across the placenta and yolk sac or is absorbed from colostrum and milk by the small intestine of the suckling newborn. In both cases apical to basolateral transepithelial transport of IgG is thought to be mediated by FcRn, an IgG Fc receptor with homology to MHC class I antigens. We have now expressed the human FcRn in polarized MDCK cells and analyzed the intracellular routing of the receptor. FcRn showed a predominant intracellular localization at steady state. Newly synthesized FcRn was delivered in a non-vectorial fashion to both the apical and basolateral surfaces of MDCK cell monolayers. Following internalization from the apical or basolateral domain, the receptor transcytosed to the opposite surface. These findings provide direct evidence for the transepithelial transport function of FcRn and indicate that the receptor undergoes multiple rounds of transcytosis. PMID- 10381386 TI - Vinblastine induces an interaction between FtsZ and tubulin in mammalian cells. AB - The Escherichia coli cell division protein FtsZ was expressed in Chinese hamster ovary cells, where it formed a striking array of dots that were independent of the mammalian cytoskeleton. Although FtsZ appears to be a bacterial homolog of tubulin, its expression had no detectable effects on the microtubule network or cell growth. However, treatment of the cells with vinblastine at concentrations that caused microtubule disassembly rapidly induced a network of FtsZ filaments that grew from and connected the dots, suggesting that the dots are an active storage form of FtsZ. Cells producing FtsZ also exhibited vinblastine- and calcium-resistant tubulin polymers that colocalized with the FtsZ network. The FtsZ polymers could be selectively disassembled, indicating that the two proteins were not copolymerized. The vinblastine effects were readily reversible by washing out the drug or by treating the cells with the vinblastine competitor, maytansine. These results demonstrate that FtsZ assembly can occur in the absence of bacterial chaperones or cofactors, that FtsZ and tubulin do not copolymerize, and that tubulin-vinblastine complexes have an enhanced ability to interact with FtsZ. PMID- 10381387 TI - Asymmetry of the spindle pole bodies and spg1p GAP segregation during mitosis in fission yeast. AB - In the fission yeast Schizosaccharomyces pombe, the onset of septum formation is induced by a signal transduction network involving several protein kinases and a GTPase switch. One of the roles of the spg1p GTPase is to localise the cdc7p protein kinase to the poles of the mitotic spindle, from where the onset of septation is thought to be signalled at the end of mitosis. Immunofluorescence studies have shown that cdc7p is located on both spindle pole bodies early in mitosis, but only on one during the later stages of anaphase. This is mediated by inactivation of spg1p on one pole before the other. The GAP for spg1p is a complex of two proteins, cdc16p and byr4p. Localisation of cdc16p and byr4p by indirect immunofluorescence during the mitotic cell cycle showed that both proteins are present on the spindle pole body in interphase cells. During mitosis, byr4p is seen first on both poles of the spindle, then on only one. This occurs prior to cdc7p becoming asymmetric. In contrast, the signal due to cdc16p decreases to a low level during early mitosis, before being seen strongly on the same pole as byr4p. Double staining indicates that this is the opposite pole to that which retains cdc7p in late anaphase. Examination of the effect of inactivating cdc16p at various stages of the cell cycle suggests that cdc16p, together with cdc2p plays a role in restraining septum formation during interphase. The asymmetric inactivation of spg1p is mediated by recruitment of the cdc16p-byr4p GAP to one of the poles of the spindle before the other, and the asymmetry of the spindle pole bodies may be established early during mitosis. Moreover, the spindle pole bodies appear to be non-equivalent even after division has been completed. PMID- 10381388 TI - The role of anillin in meiotic cytokinesis of Drosophila males. AB - Anillin is a 190 kDa actin-binding protein that concentrates in the leading edges of furrow canals during Drosophila cellularization and in the cleavage furrow of both somatic and meiotic cells. We analyzed anillin behavior during D. melanogaster spermatogenesis, and focused on the relationships between this protein and the F-actin enriched structures. In meiotic anaphases anillin concentrates in a narrow band around the cell equator. Cytological analysis of wild-type meiosis and examination of mutants defective in contractile ring assembly (chickadee and KLP3A), revealed that the formation of the anillin cortical band occurs before, and does not require the assembly of the F-actin based contractile ring. However, once the acto-myosin ring is assembled, the anillin band precisely colocalizes with this cytokinetic structure, accompanying its contraction throughout anaphase and telophase. In chickadee and KLP3A mutant ana-telophases the cortical anillin band fails to constrict, indicating that its contraction is normally driven by the cytokinetic ring. These findings, coupled with the analysis of anillin behavior in twinstar mutants, suggested a model on the role of anillin during cytokinesis. During anaphase anillin would concentrate in the cleavage furrow before the assembly of the contractile ring, binding the equatorial cortex, perhaps through its carboxy-terminal pleckstrin homology (PH) domain. Anillin would then interact with the actin filaments of the acto-myosin ring through its actin-binding domain, anchoring the contractile ring to the plasma membrane throughout cytokinesis. PMID- 10381389 TI - Integrin (alpha) and beta subunit contribution to the kinetic properties of (alpha)2beta1 collagen receptors on human keratinocytes analyzed under hydrodynamic conditions. AB - The adhesion of keratinocytes to type I collagen or laminin 5 was studied in a laminar flow chamber. These experiments provided an insight into the binding kinetics of integrins in their natural environment and the effects of monoclonal antibodies specific for (alpha) and beta chains. Cells driven by a force too low to alter the natural lifetime of a single bond displayed multiple arrests. Studying the frequency and duration of these arrests yielded fairly direct information on the rate of bond formation (on-rate) and dissociation (off-rate). Off-rate values obtained on collagen or laminin 5 (0.06 seconds-1) were tenfold lower than values determined on selectins. Bond stability was strongly regulated by anti-beta1 chain antibodies since the off-rate was decreased sixfold by activating antibody TS2/16 and increased fivefold by inhibitory antibodies Lia1/2 or P4C10, whereas neutral antibody K20 had no effect on this parameter. Binding frequencies were not significantly changed by all these antibodies. In contrast, both binding frequency and off-rate were altered by antibodies specific for the (alpha)2 chain, suggesting that these antibodies interfered with ligand recognition and also with the ligand-beta1 chain interactions responsible for bond stabilization. The latter hypothesis was supported by the finding that the partial alteration of (alpha)2 chain function by inhibiting antibodies was corrected by anti-beta1 chain antibody TS2/16. These results could not be ascribed to allosteric changes of the functional region of beta1 integrin subunits regulated by TS2/16 since there was no competition between the binding of TS2/16 and anti-(alpha)2 chain antibodies. Interpreted within the framework of current concepts of integrin-ligand binding topology, these data suggest that ligand-alpha chain interactions may be qualitatively important in ligand recognition and also influence the formation of the ligand-beta1 subunit bonding involved in stabilization of the ligand-integrin complex by regulating its dissociation rate. PMID- 10381390 TI - A single external enzyme confers alternative NADH:ubiquinone oxidoreductase activity in Yarrowia lipolytica. AB - NADH:ubiquinone oxidoreductases catalyse the first step within the diverse pathways of mitochondrial NADH oxidation. In addition to the energy-conserving form commonly called complex I, fungi and plants contain much simpler alternative NADH:ubiquinone oxido-reductases that catalyze the same reaction but do not translocate protons across the inner mitochondrial membrane. Little is known about the distribution and function of these enzymes. We have identified YLNDH2 as the only gene encoding an alternative NADH:ubiquinone oxidoreductase (NDH2) in the obligate aerobic yeast Yarrowia lipolytica. Cells carrying a deletion of YLNDH2 were fully viable; full inhibition by piericidin A indicated that complex I activity was the sole NADH:ubiquinone oxidoreductase activity left in the deletion strains. Studies with intact mitochondria revealed that NDH2 in Y. lipolytica is oriented towards the external face of the mitochondrial inner membrane. This is in contrast to the situation seen in Saccharomyces cerevisiae, Neurospora crassa and in green plants, where internal alternative NADH:ubiquinone oxidoreductases have been reported. Phylogenetic analysis of known NADH:ubiquinone oxidoreductases suggests that during evolution conversion of an ancestral external alternative NADH:ubiquinone oxidoreductase to an internal enzyme may have paved the way for the loss of complex I in fermenting yeasts like S. cerevisiae. PMID- 10381391 TI - Tau regulates the attachment/detachment but not the speed of motors in microtubule-dependent transport of single vesicles and organelles. AB - We have performed a real time analysis of fluorescence-tagged vesicle and mitochondria movement in living CHO cells transfected with microtubule-associated protein tau or its microtubule-binding domain. tau does not alter the speed of moving vesicles, but it affects the frequencies of attachment and detachment to the microtubule tracks. Thus, tau decreases the run lengths both for plus-end and minus-end directed motion to an equal extent. Reversals from minus-end to plus end directed movement of single vesicles are strongly reduced by tau, but reversals in the opposite direction (plus to minus) are not. Analogous effects are observed with the transport of mitochondria and even with that of vimentin intermediate filaments. The net effect is a directional bias in the minus-end direction of microtubules which leads to the retraction of mitochondria or vimentin IFs towards the cell center. The data suggest that tau can control intracellular trafficking by affecting the attachment and detachment cycle of the motors, in particular by reducing the attachment of kinesin to microtubules, whereas the movement itself is unaffected. PMID- 10381392 TI - Nuclear pore localization and nucleocytoplasmic transport of eIF-5A: evidence for direct interaction with the export receptor CRM1. AB - Eukaryotic initiation factor 5A (eIF-5A) is the only cellular protein known to contain the unusual amino acid hypusine. The exact in vivo function of eIF-5A, however, is to date unknown. The finding that eIF-5A is an essential cofactor of the human immunodeficiency virus type 1 (HIV-1) Rev RNA transport factor suggested that eIF-5A is part of a specific nuclear export pathway. In this study we used indirect immunofluorescence and immunogold electron microscopy to demonstrate that eIF-5A accumulates at nuclear pore-associated intranuclear filaments in mammalian cells and Xenopus oocytes. We are able to show that eIF-5A interacts with the general nuclear export receptor, CRM1. Furthermore, microinjection studies in somatic cells revealed that eIF-5A is transported from the nucleus to the cytoplasm, and that this nuclear export is blocked by leptomycin B. Our data demonstrate that eIF-5A is a nucleocytoplasmic shuttle protein. PMID- 10381393 TI - Functionally homologous DNA replication genes in fission and budding yeast. AB - The cdc18(+) gene of the fission yeast Schizosaccharomyces pombe is involved in the initiation of DNA replication as well as in coupling the S phase to mitosis. In this work, we show that the Saccharomyces cerevisiae CDC6 gene complements cdc18-K46 ts and cdc18 deletion mutant S. pombe strains. The budding yeast gene suppresses both the initiation and the checkpoint defects associated with the lack of cdc18(+). The Cdc6 protein interacts in vivo with Cdc2 kinase complexes. Interestingly, Cdc6 is an in vitro substrate for Cdc13/Cdc2 and Cig1/Cdc2, but not for Cig2/Cdc2-associated kinases. Overexpression of Cdc6 in fission yeast induces multiple rounds of S-phase in the absence of mitosis and cell division. This CDC6-dependent continuous DNA synthesis phenotype is independent of the presence of a functional cdc18(+) gene product and, significantly, requires only Cig2/Cdc2-associated kinase activity. Finally, these S. pombe over-replicating cells do not require any protein synthesis other than that of Cdc6. Our data strongly suggest that CDC6 and cdc18(+) are functional homologues and also support the idea that controls restricting genome duplication diverge in fission and budding yeast. PMID- 10381394 TI - Innexin-3 forms connexin-like intercellular channels. AB - Innexins comprise a large family of genes that are believed to encode invertebrate gap junction channel-forming proteins. However, only two Drosophila innexins have been directly tested for the ability to form intercellular channels and only one of those was active. Here we tested the ability of Caenorhabditis elegans family members INX-3 and EAT-5 to form intercellular channels between paired Xenopus oocytes. We show that expression of INX-3 but not EAT-5, induces electrical coupling between the oocyte pairs. In addition, analysis of INX-3 voltage and pH gating reveals a striking degree of conservation in the functional properties of connexin and innnexin channels. These data strongly support the idea that innexin genes encode intercellular channels. PMID- 10381395 TI - p53-independent apoptosis induced by muscle differentiation stimuli in polyomavirus large T-expressing myoblasts. AB - Abnormal proliferation signals, driven by cellular or viral oncogenes, can result in the induction of apoptosis under sub-optimal cell growth conditions. The tumor suppressor p53 plays a central role in mediating oncogene-induced apoptosis, therefore transformed cells lacking p53 are generally resistant to apoptosis promoting treatments. In a previous work we have reported that the expression of polyomavirus large T antigen causes apoptosis in differentiating myoblasts and that this phenomenon is dependent on the onset of muscle differentiation in the absence of a correct cell cycle arrest. Here we report that polyomavirus large T increases the levels and activity of p53, but these alterations are not involved in the apoptotic mechanism. Apoptosis in polyomavirus large T-expressing myoblasts is not prevented by the expression of a p53 dominant-negative mutant nor it is increased by p53 over-expression. Moreover, forced differentiation induced through the over-expression of the muscle regulatory factor MyoD, leads to apoptosis without altering p53 function and, more significantly, even in a p53 null background. Our results indicate that apoptosis induced by the activation of muscle differentiation pathways in oncogene-expressing cells can occur in a p53 independent manner. PMID- 10381396 TI - 4-hydroxynonenal triggers an epidermal growth factor receptor-linked signal pathway for growth inhibition. AB - Lipid peroxidation has been implicated in the pathogenesis of various diseases. As a major product of membrane lipid peroxidation, 4-hydroxynonenal (HNE) appears after various kinds of oxidative stress, and is known to induce cell growth inhibition. We here analysed the HNE-mediated signal transduction cascade for the growth inhibition of human epidermoid carcinoma A431 cells. HNE dose-dependently induced phosphorylation of multiple cellular proteins including epidermal growth factor receptor (EGFR) in A431 cells, and rapidly upregulated the catalytic actions of EGFR for autophosphorylation and for phosphorylation of casein as an exogenous substrate. Immunoblot analysis by use of HNE-specific antibody demonstrated the binding of HNE to EGFR along with its activation. This binding, which did not induce cross-linking of EGFR, caused a capping of the receptor on the cell surface which mimicked the capping induced by EGF. Phosphorylation and activation of EGFR were followed by phosphorylation of adaptor protein Shc and activation of MAP kinase. Both genistein as a wide spectrum protein tyrosine kinase inhibitor and AG1478 as a specific EGFR tyrosine phosphorylation blocker inhibited activation of EGFR and MAP kinase by HNE. The same inhibitors prevented HNE-mediated growth inhibition, suggesting a close linkage between EGFR/MAP kinase activation and growth inhibition after exposure to HNE. Our results suggest that EGFR may be one of the primary targets of HNE for an oxidative stress-linked cell growth inhibition. PMID- 10381397 TI - Chloromethyl-X-rosamine (MitoTracker Red) photosensitises mitochondria and induces apoptosis in intact human cells. AB - We report that chloromethyl-X-rosamine (MitoTracker Red), a mitochondrion selective fluorescent probe, has a strong photosensitising action. Photoirradiation of intact cells loaded with chloromethyl-X-rosamine induces depolarisation of the inner mitochondrial membrane and swelling of mitochondria, subsequently resulting in apoptosis. We have studied human osteosarcoma 143B TK (rho+) cells and the derived (rho)0 206 cell line devoid of mitochondrial DNA. Colony formation tests revealed that chloromethyl-X-rosamine itself has no toxicity to either cell line in the concentration range 100-250 nM (unless photoirradiated). Chloromethyl-X-rosamine has potent phototoxicity such that almost quantitative cell killing was achieved at light doses of >2 J/cm2. These photodamaged cells initially showed swollen degenerative mitochondria and, later, uptake of propidium iodide in their apoptotic nuclei was observed. When cells were loaded with chloromethyl-X-rosamine (100 nM) and imaged by laser scanning confocal microscopy, photoirradiation by the laser beam under routine scanning conditions was sufficient to induce mitochondrial damage in both cell lines. This was evidenced by a rapid decrease of fluorescence intensity of co-loaded rhodamine 123 (indicative of mitochondrial depolarisation). Globular swelling of mitochondria took place within 15 minutes, imaged by the residual fluorescence of chloromethyl-X-rosamine itself, which also markedly decreased in intensity after imaging. Mitochondrial membrane depolarisation of cells loaded with chloromethyl X-rosamine after photoirradiation using a measured dose of visible light was independently confirmed in 143B TK- and (rho)0 206 cells, by the significant decrease in uptake into cells of [3H]methyltriphenylphosphonium ions. Photoactivation of chloromethyl-X-rosamine in 143B TK-(rho+) cells, whose mitochondria had previously been loaded with calcein, caused rapid release of the mitochondrially entrapped calcein into the cytosol and nucleus. This major change in permeability of the mitochondrial inner membrane could not be prevented by cyclosporin A. Immunohistochemical study of cytochrome c revealed its diffuse redistribution into the cytoplasm in chloromethyl-X-rosamine-loaded cells after irradiation, as opposed to its specific mitochondrial localisation in non irradiated cells. As a photosensitiser specifically targeted to mitochondria, and also a reporter of membrane potential and morphology, chloromethyl-X-rosamine may provide versatile new applications in studies of mitochondrial roles in cell death. PMID- 10381398 TI - Cell surface membrane homeostasis and intracellular membrane traffic balance in mouse L929 cells. AB - We have developed a simple method for synchronizing L929 mouse fibroblasts. Cultured as monolayers, these cells stop growing at confluency and arrest at the end of the G1 phase. Upon seeding at low density, they enter the S phase simultaneously. Using these cells we then looked at the evolution of the surface membrane area during the cell cycle using the fluorescence membrane probe TMA DPH. In contact with cells, this probe partitions between the membrane (probe fluorescent) and the external medium (non-fluorescent), delivering a signal proportional to the membrane area. This area was constant until just before mitosis, when it increased at once. With the same probe as an endocytic marker, we examined how this membrane homeostasis could be consistent with intracellular membrane trafficking. The study was limited to one selected period of the cell cycle (6-9 hours). We observed that 14% of the membrane endocytosed was not recycled, but was replaced at the cell surface by newly formed membrane from biosynthetic pathways. Brefeldin A modified the membrane traffic, but not the overall membrane homeostasis. The results are discussed in the framework of a maturation model. PMID- 10381399 TI - Maternal histone deacetylase is accumulated in the nuclei of Xenopus oocytes as protein complexes with potential enzyme activity. AB - Reversible acetylation of core histones plays an important regulatory role in transcription and replication of chromatin. The acetylation status of chromatin is determined by the equilibrium between activities of histone acetyltransferases (HATs) and histone deacetylases (HDACs). The Xenopus protein HDACm shows sequence homology to other putative histone deacetylases, but its mRNA is expressed only during early development. Both HDACm protein and acetylated non-chromosomal histones are accumulated in developing oocytes, indicating that the key components for histone deposition into new chromatin during blastula formation are in place by the end of oogenesis. Here we show that the 57 kDa HDACm protein undergoes steady accumulation in the nucleus, where it is organized in a multiprotein complex of approx. 300 kDa. A second, major component of the nuclear complex is the retinoblastoma-associated protein p48 (RbAp48/46), which may be used as an adaptor to contact acetylated histones in newly assembled chromatin. The nuclear complex has HDAC activity that is sensitive to trichostatin A, zinc ions and phosphatase treatment. The 57 kDa protein serves as a marker for total HDAC activity throughout oogenesis and early embryogenesis. The active HDACm complex and its acetylated histone substrates appear to be kept apart until after chromatin assembly has taken place. However, recombinant HDACm, injected into the cytoplasm of oocytes, not only is translocated to the nucleus, but also is free to interact with the endogenous chromatin. PMID- 10381400 TI - Ran-GTP stabilises microtubule asters and inhibits nuclear assembly in Xenopus egg extracts. AB - Ran is an abundant GTPase of the Ras superfamily that is highly conserved in eukaryotes. In interphase cells, Ran is mainly nuclear and thought to be predominantly GTP-bound, but it is also present in the cytoplasm, probably GDP bound. This asymmetric distribution plays an important role in directing nucleocytoplasmic transport. Ran has also been implicated in cell cycle control, including the transition from mitosis to interphase when the compartmentalisation of the nucleus is established. Here, we have examined the role of Ran in this transition using a cell-free system of Xenopus egg extracts supplemented with sperm heads that provides a model for microtubule aster formation and post-M phase nuclear assembly. Ran-GTP, added as wild-type protein, a mutant defective in GTPase activity (Q69L), or generated by addition of the specific nucleotide exchange factor RCC1, stabilises large microtubule asters nucleated at the sperm centrosome, prevents the redistribution of NuMA from the aster to the nucleus and blocks chromatin decondensation. In contrast, Ran GDP does not stabilise microtubules or inhibit nuclear assembly. RanT24N and RanBP1, which oppose the generation of Ran-GTP by RCC1, arrest nuclear growth after disappearance of the aster. Ran associates with microtubule asters in egg extracts and with mitotic spindles in somatic Xenopus cells, suggesting that it may affect microtubule stability directly. These results show that Ran has a novel function in the control of microtubule stability that is clearly distinct from nucleocytoplasmic transport. The Ran GDP/GTP switch may play a role in co-ordinating changes in the structure of microtubules and the assembly of the nucleus associated with the transition from mitosis to interphase. PMID- 10381401 TI - A novel acidotropic pH indicator and its potential application in labeling acidic organelles of live cells. AB - BACKGROUND: Ratio imaging has received intensive attention in the past few decades. The growing potential of ratio imaging is significantly limited, however, by the lack of appropriate fluorescent probes, for acidic organelles in particular. The classic fluorescent dyes (such as fluoresceins, rhodamines and coumarins) are not suitable for studying acidic organelles (such as lysosomes) because their fluorescence is significantly decreased under neutral or acidic conditions. This has motivated us to develop probes that can be used in ratio imaging that are strongly fluorescent even in acidic media. RESULTS: The compound 2-(4-pyridyl)-5-((4-(2-dimethylaminoethyl-aminocarbamoyl) methoxy)phenyl)oxazole (PDMPO) was prepared and characterized as a new acidotropic dual-excitation and dual-emission pH indicator. It emits intense yellow fluorescence at lower pH and gives intense blue fluorescence at higher pH. This unique pH-dependent fluorescence property was readily explored to selectively stain lysosomes and to determine the pH of the organelle in an emission-ratio-imaging mode. PDMPO is selectively localized to lysosomes and exhibits a pH-dependent dual excitation and emission. CONCLUSIONS: PDMPO selectively labels acidic organelles (such as lysosomes) of live cells and the two distinct emission peaks can be used to monitor the pH fluctuations of live cells in ratio measurements. Additionally, the very large Stokes shift and excellent photostability of PDMPO make the compound an ideal fluorescent acidotropic probe. The unique fluorescence properties of PDMPO might give researchers a new tool with which to study acidic organelles of live cells. PMID- 10381402 TI - Probing intermolecular backbone H-bonding in serine proteinase-protein inhibitor complexes. AB - BACKGROUND: Intermolecular backbone H-bonding (N-H.O=C) is a common occurrence at the interface of protein-protein complexes. For instance, the amide NH groups of most residues in the binding loop of eglin c, a potent serine proteinase inhibitor from the leech Hirudo medicinalis, are H-bonded to the carbonyl groups of residues in the target enzyme molecules such as chymotrypsin, elastase and subtilisins. We sought to understand the energetic significance of these highly conserved backbone-backbone H-bonds in the enzyme-inhibitor complexes. RESULTS: We synthesized an array of backbone-engineered ester analogs of eglin c using native chemical ligation to yield five inhibitor proteins each containing a single backbone ester bond from P3 to P2' (i.e. -CONH-to -COO-). The structure at the ligation site (P6-P5) is essentially unaltered as shown by a high-resolution analysis of the subtilisin-BPN'-eglin c complex. The free-energy changes (DeltaDeltaGNH-->O) associated with the binding of ester analogs at P3, P1 and P2' with bovine alpha-chymotrypsin, subtilisin Carlsberg and porcine pancreatic elastase range from 0-4.5 kcal/mol. Most markedly, the NH-->O substitution at P2 not only stabilizes the inhibitor but also enhances binding to the enzymes by as much as 500-fold. CONCLUSIONS: Backbone H-bond contributions are context dependent in the enzyme-eglin c complexes. The interplay of rigidity and adaptability of the binding loop of eglin c seems to play a prominent role in defining the binding action. PMID- 10381403 TI - Reactivity of guanine at m5CpG steps in DNA: evidence for electronic effects transmitted through the base pairs. AB - BACKGROUND: Mitomycin C (MC), a DNA cross-linking and alkylating agent, targets guanines in the m5CpG sequence with 2-3-fold preference over guanines in unmethylated CpG. Benzo[a]pyrenediolepoxide (BPDE) and several other aromatic carcinogens form guanine adducts with an identical selectivity for m5CpG, and in certain cancers G to T transversion mutation 'hotspots' in the p53 tumor suppressor gene are more frequent at this sequence than at guanines in other sequences. MC appears suitable to probe the general mechanism of this selectivity. RESULTS: A 162-bp DNA fragment containing C, m5C or f5C (5-fluoro cytosine) at all cytosine positions was cross-linked by MC at guanines in CpG steps. The extent of cross-linking increased in the order f5C < C < m5C. Monoalkylation or cross-linking of duplex 12-mer oligonucleotides containing a single CpG, f5CpG or m5CpG step gave yields of adducts that increased in the same order. The rates showed a correlation with the Hammett sigma constant of the methyl and fluoro substituents of the cytosine. Only the base-pair cytosine substituent influenced reactivity of guanine. CONCLUSIONS: The 2-amino group of guanine in the m5CpG sequence of DNA has a greater nucleophilic reactivity with mitomycin than CpG. Evidence is presented for a novel mechanism: transmission of the electron-donating effect of the 5-methyl substituent of the cytosine to guanine through H-bonding of the m5C.G base pair. The results explain the enhanced reaction of BPDE at m5CpG in DNA and the origin of G-T mutational hotspots in the p53 gene in cancer. PMID- 10381406 TI - A highly membrane-active peptide in Flock House virus: implications for the mechanism of nodavirus infection. AB - BACKGROUND: Nodaviruses are among the simplest animal viruses, and are therefore attractive systems for deconvoluting core viral processes such as assembly, infection and uncoating. Membrane translocation of the single-stranded RNA genome of nodaviruses has been proposed to be mediated by direct lipid-protein interactions between a post-assembly autocatalytic cleavage product from the capsomere and the target membrane. To probe the validity of this hypothesis, we have synthesized a 21-residue Met-->Nle (norleucine) variant of the amino terminal helical domain (denoted here as gamma1) of the cleavage peptide in Flock House nodavirus (FHV) and studied its ability to alter membrane structure and function. RESULTS: The synthetic peptide gamma1 increases membrane permeability to hydrophilic solutes, as judged by fluorescence experiments with liposome encapsulated dyes and ion-conductance measurements. Furthermore, peptide orientation and location within lipid bilayers was determined using tryptophan fluorescence-quenching experiments and attenuated total reflectance infrared spectroscopy. CONCLUSIONS: The helical domain of the FHV cleavage product partitions spontaneously into lipid bilayers and increases membrane permeability, consistent with the postulated mechanism for viral genome translocation. The existence of a membrane-binding domain in the FHV cleavage sequence suggests peptide-triggered disruption of the endosomal membrane as a prelude to viral uncoating in the host cytoplasm. A model for this interaction is proposed. PMID- 10381405 TI - The effects of secondary structure and O2 on the formation of direct strand breaks upon UV irradiation of 5-bromodeoxyuridine-containing oligonucleotides. AB - BACKGROUND: 5-Bromodeoxyuridine is a radiosensitizing agent that is currently being evaluated in clinical trials as an adjuvant in the treatment of a variety of cancers. gamma-Radiolysis and UV irradiation of oligonucleotides containing 5 bromodeoxyuridine result in the formation of direct strand breaks at the 5' adjacent nucleotide by oxidation of the respective deoxyribose. We investigated the effects of DNA secondary structure and O2 on the induction of direct strand breaks in 5-bromodeoxyuridine-containing oligonucleotides. RESULTS: The efficiency of direct strand break formation in duplex DNA is dependent upon O2 and results in fragments containing 3'-phosphate and the labile 3' ketodeoxyadenosine termini. The ratio of the 3'-termini is also dependent upon O2 and structure. Deuterium product isotope effects and tritium-transfer studies indicate that hydrogen-atom abstraction from the C1'- and C2'-positions occurs in an O2- and structure-dependent manner. CONCLUSIONS: The reaction mechanisms by which DNA containing 5-bromodeoxyuridine is sensitized to damage by UV irradiation are dependent upon whether the substrate is hybridized and upon the presence or absence of O2. Oxygen reduces the efficiency of direct strand break formation in duplex DNA, but does not affect the overall strand damage. It is proposed that the sigma radical abstracts hydrogen atoms from the C1'- and C2' positions of the 5'-adjacent deoxyribose moiety, whereas the nucleobase peroxyl radical selectively abstracts the C1'-hydrogen atom from this site. This is the second example of DNA damage amplification by a nucleobase peroxyl radical, and might be indicative of a general reaction pattern for this family of reactive intermediates. PMID- 10381404 TI - Mutational and structural analyses of the regulatory protein B of soluble methane monooxygenase from Methylococcus capsulatus (Bath). AB - BACKGROUND: The soluble methane monooxygenase (sMMO) system in methanotrophic bacteria uses three protein components to catalyze the selective oxidation of methane to methanol. The coupling protein B (MMOB) both activates the carboxylate bridged diiron center in the hydroxylase (MMOH) for substrate oxidation and couples the reaction to electron transfer from NADH through the sMMO reductase. Although the X-ray structure of the hydroxylase is known, little structural information is available regarding protein B. RESULTS: Wild-type protein B from Methylococcus capsulatus (Bath) is very susceptible to degradation. The triple mutant protein B, Gly10-->Ala, Gly13-->Gln, Gly16-->Ala is resistant to degradation. Analyzing wild-type and mutant forms of protein B using size exclusion chromatography and circular dichroism spectroscopy suggests that the amino terminus of MMOB (Ser1-Ala25) is responsible for the proteolytic sensitivity and unusual mobility of the protein. We used the stable triple glycine protein B mutant to generate an affinity column for the hydroxylase and investigated the interaction between MMOH and MMOB. These results suggest the interaction is dominated by hydrophobic contacts. CONCLUSIONS: A structural model is presented for protein B that explains both its proclivity for degradation and its anomalous behavior during size exclusion chromatography. The model is consistent with previously published biophysical data, including the NMR structure of the phenol hydroxylase regulatory protein P2. Furthermore, this model allows for detailed and testable predictions about the structure of protein B and the role of proposed recognition sites for the hydroxylase. PMID- 10381407 TI - Lovastatin biosynthesis in Aspergillus terreus: characterization of blocked mutants, enzyme activities and a multifunctional polyketide synthase gene. AB - BACKGROUND: Lovastatin, an HMG-CoA reductase inhibitor produced by the fungus Aspergillus terreus, is composed of two polyketide chains. One is a nonaketide that undergoes cyclization to a hexahydronaphthalene ring system and the other is a simple diketide, 2-methylbutyrate. Fungal polyketide synthase (PKS) systems are of great interest and their genetic manipulation should lead to novel compounds. RESULTS: An A. terreus mutant (BX102) was isolated that could not synthesize the nonaketide portion of lovastatin and was missing a approximately 250 kDa polypeptide normally present under conditions of lovastatin production. Other mutants produced lovastatin intermediates without the methylbutyryl sidechain and were missing a polypeptide of approximately 220 kDa. The PKS inhibitor cerulenin reacted covalently with both polypeptides. Antiserum raised against the approximately 250 kDa polypeptide was used to isolate the corresponding gene, which complemented the BX102 mutation. The gene encodes a polypeptide of 269 kDa containing catalytic domains typical of vertebrate fatty acid and fungal PKSs, plus two additional domains not previously seen in PKSs: a centrally located methyltransferase domain and a peptide synthetase elongation domain at the carboxyl terminus. CONCLUSIONS: The results show that the nonaketide and diketide portions of lovastatin are synthesized by separate large multifunctional PKSs. Elucidation of the primary structure of the PKS that forms the lovastatin nonaketide, as well as characterization of blocked mutants, provides new details of lovastatin biosynthesis. PMID- 10381408 TI - Hydrogen tunneling in biology. AB - The mechanistic details of hydrogen transfer in biological systems are not fully understood. The traditional approach has been to use semiclassical transition state theory. This theory cannot explain many experimental findings, however, so different approaches that emphasize the importance of quantum mechanics and dynamic effects should also be considered. PMID- 10381409 TI - Catalysis and specificity in enzymatic glycoside hydrolysis: a 2,5B conformation for the glycosyl-enzyme intermediate revealed by the structure of the Bacillus agaradhaerens family 11 xylanase. AB - BACKGROUND: The enzymatic hydrolysis of glycosides involves the formation and subsequent breakdown of a covalent glycosyl-enzyme intermediate via oxocarbenium ion-like transition states. The covalent intermediate may be trapped on-enzyme using 2-fluoro-substituted glycosides, which provide details of the intermediate conformation and noncovalent interactions between enzyme and oligosaccharide. Xylanases are important in industrial applications - in the pulp and paper industry, pretreating wood with xylanases decreases the amount of chlorine containing chemicals used. Xylanases are structurally similar to cellulases but differ in their specificity for xylose-based, versus glucose-based, substrates. RESULTS: The structure of the family 11 xylanase, Xyl11, from Bacillus agaradhaerens has been solved using X-ray crystallography in both native and xylobiosyl-enzyme intermediate forms at 1.78 A and 2.0 A resolution, respectively. The covalent glycosyl-enzyme intermediate has been trapped using a 2-fluoro-2-deoxy substrate with a good leaving group. Unlike covalent intermediate structures for glycoside hydrolases from other families, the covalent glycosyl-enzyme intermediate in family 11 adopts an unusual 2,5B conformation. CONCLUSIONS: The 2,5B conformation found for the alpha-linked xylobiosyl-enzyme intermediate of Xyl11, unlike the 4C1 chair conformation observed for other systems, is consistent with the stereochemical constraints required of the oxocarbenium-ion-like transition state. Comparison of the Xyl11 covalent glycosyl-enzyme intermediate with the equivalent structure for the related family 12 endoglucanase, CelB, from Streptomyces lividans reveals the likely determinants for substrate specificity in this clan of glycoside hydrolases. PMID- 10381410 TI - Presenilins in their infancy. AB - Familial forms of Alzheimer's disease are caused by mutations in the genes encoding the presenilins, which are integral membrane proteins. Presenilins have been shown to interact with beta-amyloid precursor proteins and Notch receptors. Several recent studies have examined the role of presenilins in Notch processing. PMID- 10381479 TI - Changing of the guard PMID- 10381478 TI - From nottingham to nijmegen PMID- 10381480 TI - Risk of post-discharge venous thromboembolism in patients with rheumatoid arthritis undergoing knee or hip arthroplasty. Is prolonged thromboprophylaxis warranted or dangerous? PMID- 10381481 TI - A concealed cause of recurrent renal failure in a patient with juvenile chronic arthritis. PMID- 10381483 TI - Bone densitometry in clinical practice PMID- 10381482 TI - Lumbar osteotomy for correction of thoracolumbar kyphotic deformity in ankylosing spondylitis. A structured review of three methods of treatment. AB - OBJECTIVES: Three operative techniques have been described to correct thoracolumbar kyphotic deformity (TLKD) resulting from ankylosing spondylitis (AS) at the level of the lumbar spine: opening wedge osteotomy, polysegmental wedge osteotomies, and closing wedge osteotomy. Little knowledge exists on the indication for, and outcome of these corrective lumbar osteotomies. METHODS: A structured review of the medical literature was performed. RESULTS: A search of the literature revealed 856 patients reported in 41 articles published between 1945 and 1998. The mean age at time of operation was 41 years, male-female ratio 7.5 to 1. In 451 patients an open wedge osteotomy was performed. Polysegmental wedge osteotomies were performed in 249 patients and a closing wedge osteotomy in 156 patients. Most of the studies primarily focus on the surgical technique. Technical outcome data were poorly reported. Sixteen reports, including 523 patients, met the inclusion criteria of this study, and could be analysed for technical outcome data. The average correction achieved with each surgical techniques ranged from 37 to 40 degrees. Loss of correction was mainly reported in patients treated by open wedge osteotomy and polysegmental wedge osteotomies. Neurological complications were reported in all three techniques. The perioperative mortality was 4%. Pulmonary, cardiac and intestinal problems were found to be the major cause of fatal complications. CONCLUSION: Lumbar osteotomy for correction of TLKD resulting from AS is a major surgery. The indication for these lumbar osteotomies as well as the degree of correction in the lumbar spine has not yet been established. Furthermore, there is a need for a generally accepted clinical score that encompasses accurate preoperative and postoperative assessment of the spinal deformity. The results of this review suggest that the data from the literature are not suitable for decision making with regard to surgical treatment of TLKD resulting from AS. PMID- 10381484 TI - A clinical and serological comparison of group A versus non-group A streptococcal reactive arthritis and throat culture negative cases of post-streptococcal reactive arthritis. AB - OBJECTIVE: To identify clinical and serological differences of patients with reactive arthritis after infection with Lancefield group A beta-haemolytic streptococci (GAS), compared with non-group A-that is, group C or G streptococci (NGAS:GCS/GGS), and a group of culture negative or unidentified streptococci (GUS). METHODS: A prospective study of consecutive patients with reactive arthritis after serologically or culture confirmed infection with beta-haemolytic streptococci, presenting to the outpatient department of rheumatology from January 1992 until January 1998. Alternative causes for reactive arthritis were excluded. Main outcome measures were clinical and serological characteristics including antistreptolysine-O (ASO) and antideoxyribonuclease-B (antiDNase-B) antibody titres. RESULTS: 41 patients (female/male ratio 22/19; mean (SD) age 38 (13) years) with reactive arthritis were included. Culture of throat swab was positive in 13 cases (32%): 6 (15%) GAS, 7 NGAS (17%), that is, 5 (12%) GCS, 2 (5%) GGS. In 28 cases throat culture remained negative resulting in a group of unidentified streptococci; antibiotic pre-treatment had been given by the general practitioner in 18 cases (64%). Arthritis was non-migratory, the number of arthritic joints in GAS and NGAS was similar, whereas in NGAS patients fewer joints were involved than in GUS: mean (SEM) 36 swollen joint index: 3.3 (1.0) in NGAS v 5.6 (1.0) in GUS (p<0.005); 28 swollen joint index: 2.9 (1.0) in NGAS v 4.3 (0.8) in GUS (p<0.05). Extra-articular manifestations-that is, erythema nodosum/ multiforme, AV conduction block or hepatitis-were observed after GAS or GUS infection, but not after NGAS infection. ASO and/or antiDNase-B rose significantly in all patients. The maximal titres for ASO and antiDNase-B in 41 PSRA patients were: mean (SEM) 1242 (232) U/l and 890 (100) U/l respectively; the maximal ASO titres were similar in the three groups: mean (SEM) 1125 (185) in GAS, 625 (160) in NGAS (GAS v NGAS: p=0.17), and 1430 (320) U/l in GUS (NGAS v GUS: p=0.10). AntiDNase-B titres were: mean (SEM) 1075 (180) in GAS, 375 (105) in NGAS (GAS v NGAS: p<0.01), and 995 (125) U/l in GUS (NGAS v GUS: p<0.005). ASO: antiDNase-B ratios were: mean (SEM) 0.89 (0.21) in GAS, 2.60 (0.76) in NGAS (GAS v NGAS: p<0.05), and 1.43 (0.28) in GUS (NGAS v GUS: p=0.12). CONCLUSION: Post streptococcal reactive arthritis occurs not infrequently. Differentiation of PSRA based on the causative streptococcal strain is frequently thwarted by negative throat cultures. Sometimes extra-articular manifestations are present that exclude NGAS as the causative organism. Serologically, lower antiDNase-B titres may be indicative for primary NGAS infection; the ASO/antiDNase-B ratio may be of additive value for differentiation in cases of a negative throat culture: the higher ASO/antiDNase-B ratios suggesting primary NGAS infection. In reactive arthritis, serological monitoring consisting of a simultaneous titration of antiDNase-B and ASO, seems to be of clinical importance to trace GAS induced cases, especially when throat cultures remain negative. PMID- 10381485 TI - Sicca symptoms, saliva and tear production, and disease variables in 636 patients with rheumatoid arthritis. AB - OBJECTIVES: (1) To estimate the prevalence of ocular and oral sicca symptoms (SISY) or reduced saliva and tear production; (2) to relate SISY and sicca signs to measures of disease activity, damage, and health status; and (3) to examine the relation between symptoms and objective signs of tear and saliva production in a large sample of representative patients with rheumatoid arthritis (RA). METHODS: From an unselective county RA register 636 patients (age 20-70 years) were examined with Schirmer-I test (ST), unstimulated whole saliva (UWS), questions on SISY and measures of disease activity, damage and health status. RESULTS: Ocular sicca symptoms were reported in 38%, oral sicca symptoms in 50%, and a combination of both in 27%. Reduced tear production was present in 29%, and reduced saliva production in 17%. The minimum frequency of secondary Sjogren's syndrome was 7%. Measurements of exocrine disease manifestations were to variable extents bivariately correlated to disease activity measures, physical disability, pain, fatigue, and use of xerogenic drugs, but were not related to deformed joint count. Multivariate analyses revealed significant associations between disease activity and reduced saliva production. Only weak associations between SISY and tear or saliva production were observed. CONCLUSION: SISY, reduced tear and saliva production were frequent extra-articular manifestations in RA, but were only weakly intercorrelated. High disease activity and at least two SISY were independent predictors of reduced saliva production, but ocular and oral dryness did not seem to be closely related to disease duration, disease activity, damage or health status. PMID- 10381486 TI - Clinical follow up study of 87 patients with sicca symptoms (dryness of eyes or mouth, or both). AB - OBJECTIVE: To assess the prognosis of patients with sicca symptoms and to identify the clinical and immunological factors that most sensitively predict the later development of primary Sjogren's syndrome (SS) or other connective tissue diseases. METHODS: Eighty seven patients (72 female, 15 male) with sicca symptoms were re-evaluated after a median follow up time of 11 years (range 8-17). The clinical examination included ophthalmological examination (Schirmer's test, break up time and Rose-Bengal staining). Labial salivary gland biopsy was performed and histological findings graded according to the Chisholm-Mason scale. The immunoserological tests included determination of rheumatoid factor (RF), antinuclear antibodies (ANA), anti-extractable nuclear antigen-antibodies (ENA), serum immunoglobulins IgA, IgG, and IgM, and serum beta2-microglobulin (beta2m). RESULTS: At follow up 31 patients (36%) fulfilled modified Californian criteria (salivary flow measurements were not performed and Chisholm-Mason grades 3-4 were regarded as diagnostic histological findings) for possible or definite SS. Likewise, a significant progression of the histological findings was observed. Labial salivary gland re-biopsy was performed in 42 patients with grade 0-2 findings at baseline, progression to grades 3-4 being observed in 21 (50%) at follow up. The patients who later developed SS were at baseline significantly older (mean (SD) 52 (9) v 44 (14) years, p99%) from atopic asthmatics with 500 U/mL IFN-gamma to analyze the kinetics of mobilization and release of RANTES (0 to 240 minutes). We used subcellular fractionation, immunogold analysis, two-color confocal laser scanning microscopy (CLSM), and enzyme-linked immunosorbent assay (ELISA) to trace the movement of eosinophil-derived RANTES from intracellular stores to release. RANTES was rapidly mobilized (10 minutes) and released after 120 minutes of stimulation (80 +/- 15 pg/mL per 2 x 10(6) cells). RANTES appeared to be stored in at least two intracellular compartments: the matrix of crystalloid granules, detected by major basic protein and eosinophil peroxidase activities, and a specialized small secretory vesicle present in light membrane fractions. The extragranular RANTES was mobilized more rapidly than that of crystalloid granules during IFN-gamma stimulation. This effect was not observed in eosinophils treated with IFN-alpha, interleukin-3 (IL-3), IL-5, granulocyte-macrophage colony-stimulating factor (GM CSF), or genistein followed by IFN-gamma. Our findings suggest that RANTES may be mobilized and released by piecemeal degranulation upon stimulation, involving transport through a putative pool of small secretory vesicles. PMID- 10381495 TI - Elderly aggressive-histology non-Hodgkin's lymphoma: first-line VNCOP-B regimen experience on 350 patients. AB - Age is a risk factor and a prognostic parameter in elderly aggressive-histology non-Hodgkin's lymphoma (NHL) patients. Several adapted chemotherapeutic regimens have recently been designed and tested on elderly patients. Several of these trials have shown that older aggressive-histology NHL patients can benefit from specific and adequate treatment capable of curing a percentage of these patients. Between January 1992 and September 1997, 350 previously untreated aggressive histology NHL patients greater than 60 years of age were treated with a combination therapy including cyclophosphamide, mitoxantrone, vincristine, etoposide, bleomycin, and prednisone (VNCOP-B). Complete remission (CR) was achieved by 202 (58%) patients and partial remission (PR) by 87 (25%), whereas the remaining 61 (17%) patients were nonresponders. The overall response rate (CR + PR) was 83%. Clinical and hematologic toxicities were modest, because 71% of the patients received granulocyte colony-stimulating factor (G-CSF). The CR rates for the three age groups (60 to 69, 70 to 79, and >/=80 years) were similar: 61%, 59%, and 56%, respectively. At 5 years, the relapse-free survival rate was 65%, the overall survival rate was 49%, and the failure-free survival rate was 33%. In the multivariate analysis, prognostic factors associated with longer survival or longer relapse-free survival turned out to be localized disease stage (P =.001) and good performance status (P =.0002). Application of the International Prognostic Factor Index was significantly associated with outcome (P =.001). These data confirm on a large cohort of patients that the VNCOP-B regimen is effective in inducing good CR and relapse-free survival rates with only moderate toxic effects in elderly aggressive-histology NHL. PMID- 10381496 TI - Complete remission of t(11;17) positive acute promyelocytic leukemia induced by all-trans retinoic acid and granulocyte colony-stimulating factor. AB - The combined use of retinoic acid and chemotherapy has led to an important improvement of cure rates in acute promyelocytic leukemia. Retinoic acid forces terminal maturation of the malignant cells and this application represents the first generally accepted differentiation-based therapy in leukemia. Unfortunately, similar approaches have failed in other types of hematological malignancies suggesting that the applicability is limited to this specific subgroup of patients. This has been endorsed by the notorious lack of response in acute promyelocytic leukemia bearing the variant t(11;17) translocation. Based on the reported synergistic effects of retinoic acid and the hematopoietic growth factor granulocyte colony-stimulating factor (G-CSF), we studied maturation of t(11;17) positive leukemia cells using several combinations of retinoic acid and growth factors. In cultures with retinoic acid or G-CSF the leukemic cells did not differentiate into mature granulocytes, but striking granulocytic differentiation occurred with the combination of both agents. At relapse, the patient was treated with retinoic acid and G-CSF before reinduction chemotherapy. With retinoic acid and G-CSF treatment alone, complete granulocytic maturation of the leukemic cells occurred in vivo, followed by a complete cytogenetical and hematological remission. Bone marrow and blood became negative in fluorescense in situ hybridization analysis and semi-quantitative polymerase chain reaction showed a profound reduction of promyelocytic leukemia zinc finger-retinoic acid receptor-alpha fusion transcripts. This shows that t(11;17) positive leukemia cells are not intrinsically resistant to retinoic acid, provided that the proper costimulus is administered. These observations may encourage the investigation of combinations of all-trans retinoic acid and hematopoietic growth factors in other types of leukemia. PMID- 10381497 TI - Prothrombotic genetic risk factors in young survivors of myocardial infarction. AB - It has long been thought that an individual thrombotic tendency increases the risk of myocardial infarction, especially in young adults. Several "prothrombotic" genetic factors that may influence the individual thrombotic risk have been identified. To investigate the association between the risk of myocardial infarction at a young age and genetic factors thought to be associated with an increased tendency to thrombosis (the polymorphisms 4G/5G of the PAI-1 gene, PIA1/PIA2 of the platelet glycoprotein IIIa, C3550T of the platelet glycoprotein Ib gene, G10976A of the factor VII gene, C677T of the methylenetetrahydrofolate reductase gene, G1691A of the factor V gene, and G20210A of the prothrombin gene), we performed a case-control study evaluating 200 survivors (185 men, 15 women) of myocardial infarction who had experienced the event before the age of 45 years and 200 healthy subjects with a negative exercise test, individually matched for sex, age, and geographic origin with the cases. The presence of the PIA2 polymorphic allele was the only prothrombotic genetic factor associated with the risk of myocardial infarction at a young age. The odds ratio for carriers of the PIA2 allele compared with those of the PIA1 allele was 1.84 (95% confidence intervals (CI) 1.12 to 3.03). There was a significant interaction between the presence of the PIA2 allele and smoking: with their simultaneous presence, 46% (95% confidence intervals 11% to 81%) of premature myocardial infarctions were attributable to the interaction between the two factors. In conclusion, carrying the PIA2 polymorphic allele of platelet glycoprotein IIIa was the only genetic prothrombotic factor associated with the risk of developing myocardial infarction at a young age. The clinical expression of this genetic predisposition seems to be enhanced by smoking. PMID- 10381498 TI - Engraftment of MDR1 and NeoR gene-transduced hematopoietic cells after breast cancer chemotherapy. AB - To determine whether the multidrug resistance gene MDR1 could act as a selectable marker in human subjects, we studied engraftment of peripheral blood progenitor cells (PBPCs) transduced with either MDR1 or the bacterial NeoR gene in six breast cancer patients. This study differed from previous MDR1 gene therapy studies in that patients received only PBPCs incubated in retroviral supernatants (no nonmanipulated PBPCs were infused), transduction of PBPCs was supported with autologous bone marrow stroma without additional cytokines, and a control gene (NeoR) was used for comparison with MDR1. Transduced PBPCs were infused after high-dose alkylating agent therapy and before chemotherapy with MDR-substrate drugs. We found that hematopoietic reconstitution can occur using only PBPCs incubated ex vivo, that the MDR1 gene product may play a role in engraftment, and that chemotherapy may selectively expand MDR1 gene-transduced hematopoietic cells relative to NeoR transduced cells in some patients. PMID- 10381499 TI - Human CD34(+) cells express CXCR4 and its ligand stromal cell-derived factor-1. Implications for infection by T-cell tropic human immunodeficiency virus. AB - Human CD34(+) hematopoietic progenitor cells obtained from bone marrow (BM), umbilical cord blood (UCB), and mobilized peripheral blood (MPB) were purified and investigated for the expression of the chemokine receptor CXCR4 and its ligand, stromal cell-derived factor-1 (SDF-1). CXCR4 was found present on the cell surface of all CD34(+) cells, although it was expressed at lower density on MPB with respect to BM CD34(+) cells. Freshly isolated and in vitro-cultured CD34(+) cells also coexpressed SDF-1 mRNA, as determined by reverse transcriptase polymerase chain reaction (RT-PCR). Of interest, CD34(+)/CD38(+) committed progenitor cells, unlike primitive CD34(+)/CD38(-) cells, expressed SDF-1 mRNA. Supernatants from in vitro-cultured CD34(+) cells contained substantial (3 to 8 ng/mL) amounts of SDF-1 by enzyme-linked immunosorbent assay and induced migration of CD34(+) cells. Because CD34(+) cells express low levels of CD4, the primary receptor of the human immunodeficiency virus (HIV), and CXCR4 is a coreceptor for T-cell tropic (X4) HIV strains, we investigated the susceptibility of CD34(+) cells to infection by this subset of viruses. Lack of productive infection was almost invariably observed as determined by a conventional RT activity in culture supernatants and by real-time PCR for HIV DNA in CD34(+) cells exposed to both laboratory adapted (LAI) and primary (BON) X4 T-cell tropic HIV-1 strain. Soluble gp120 Env (sgp120) from X4 HIV-1 efficiently blocked binding of the anti-CD4 Leu3a monoclonal antibody (MoAb) to either human CD4(+) T cells or CD34(+) cells. In contrast, sgp120 interfered with an anti-CXCR4 MoAb binding to human T lymphocytes, but not to CD34(+) cells. However, CXCR4 on CD34(+) cells was downregulated by SDF-1. These results suggest that CXCR4 and its ligand SDF-1 expressed in CD34(+) progenitors may play an important role in regulating the local and systemic trafficking of these cells. Moreover, these findings suggest multiple and potentially synergistic mechanisms at the basis of the resistance of CD34(+) cells to X4 HIV infection, including their ability to produce SDF-1, and the lack of CXCR4 internalization following gp120 binding to CD4. PMID- 10381500 TI - Genetic evidence for an additional factor required for erythropoietin-induced signal transduction. AB - Erythropoietin (EPO) and its receptor (EPOR) are required for the development of mature erythrocytes. After binding of ligand, the EPOR activates a variety of signaling pathways that ultimately control cellular proliferation, survival, and specific gene expression. Although erythroid progenitors appear to be the principal EPO-responsive cell type in vivo due to the restricted expression of the EPOR, many growth factor-dependent cell lines expressing the EPOR can respond to EPO by activating many or all of these pathways. In the present study, we have identified a cellular context (the interleukin-2 [IL-2]-dependent HT-2 line) in which the EPO stimulation of the EPOR fails to support cellular proliferation, STAT-5 induction, or MAPK activation, despite efficient phosphorylation of the EPOR and JAK2 and inhibition of apoptosis after withdrawal of IL-2. Interestingly, when we fused HT-2 cells expressing the EPOR with Ba/F3 cells in a complementation assay, the resulting hybridomas proliferated and potently activated STAT-5 and MAPK in response to EPO. These data indicate that an unidentified cellular factor is needed to mediate signaling by the EPOR. Moreover, Ba/F3 cells apparently express this factor(s) and somatic fusions can, therefore, confer EPO-responsiveness to HT-2 cells that lack this factor. PMID- 10381501 TI - GATA-1 and erythropoietin cooperate to promote erythroid cell survival by regulating bcl-xL expression. AB - The transcription factor GATA-1 is essential for normal erythropoiesis. By examining in vitro-differentiated embryonic stem cells, we showed previously that in the absence of GATA-1, committed erythroid precursors fail to complete maturation and instead undergo apoptosis. The mechanisms by which GATA-1 controls cell survival are unknown. Here we report that in erythroid cells, GATA-1 strongly induces the expression of the anti-apoptotic protein bcl-xL, but not the related proteins bcl-2 and mcl-1. Consistent with a role for bcl-xL in mediating GATA-1-induced erythroid cell survival, in vitro-differentiated bcl-xL-/- embryonic stem cells fail to generate viable mature definitive erythroid cells, a phenotype resembling that of GATA-1 gene disruption. In addition, we show that erythropoietin, which is also required for erythroid cell survival, cooperates with GATA-1 to stimulate bcl-xL gene expression and to maintain erythroid cell viability during terminal maturation. Together, our data show that bcl-xL is essential for normal erythroid development and suggest a regulatory hierarchy in which bcl-xL is a critical downstream effector of GATA-1 and erythropoietin mediated signals. PMID- 10381502 TI - Expression of the thrombopoietin gene in human fetal and neonatal tissues. AB - Thrombopoietin (TPO) regulates megakaryopoiesis and platelet production. In the adult, TPO is mainly produced by the liver and the kidneys. This study focuses on fetal and neonatal TPO mRNA expression. In 26 human fetuses and preterm neonates, samples from liver, kidney, spleen, lung, and bone marrow were extracted for total RNA. We measured platelet counts, TPO serum concentrations by enzyme-linked immunosorbent assay, and TPO mRNA contents by reverse transcription/competitive polymerase chain reaction. TPO mRNA concentrations per microgram total RNA were similar in liver, spleen, and bone marrow, slightly lower in kidney, and significantly lower in lung. When related to gram tissue, TPO mRNA levels were highest in the liver. Considering the total amount of TPO mRNA produced in liver, kidney, and spleen, the liver accounted for 95.3%. No correlations between TPO mRNA expression and serum TPO concentration, blood platelet count, or gestational age were observed. In conclusion, the liver is the primary site of TPO gene expression in human fetuses and neonates. The spleen may contribute to TPO production during fetal life. Like in the adult, TPO mRNA is expressed in fetal bone marrow. PMID- 10381503 TI - Ex vivo expansion of autologous bone marrow CD34(+) cells with porcine microvascular endothelial cells results in a graft capable of rescuing lethally irradiated baboons. AB - Hematopoietic stem cell (HSC) self-renewal in vitro has been reported to result in a diminished proliferative capacity or acquisition of a homing defect that might compromise marrow repopulation. Our group has demonstrated that human HSC expanded ex vivo in the presence of porcine microvascular endothelial cells (PMVEC) retain the capacity to competitively repopulate human bone fragments implanted in severe combined immunodeficiency (SCID) mice. To further test the marrow repopulating capacity of expanded stem cells, our laboratory has established a myeloablative, fractionated total body irradiation conditioning protocol for autologous marrow transplantation in baboons. A control animal, which received no transplant, as well as two animals, which received a suboptimal number of marrow mononuclear cells, died 37, 43, and 59 days postirradiation, respectively. Immunomagnetically selected CD34(+) marrow cells from two baboons were placed in PMVEC coculture with exogenous human cytokines. After 10 days of expansion, the grafts represented a 14-fold to 22-fold increase in cell number, a 4-fold to 5-fold expansion of CD34(+) cells, a 3-fold to 4-fold increase of colony-forming unit-granulocyte-macrophage (CFU-GM), and a 12-fold to 17-fold increase of cobblestone area-forming cells (CAFC) over input. Both baboons became transfusion independent by day 23 posttransplant and achieved absolute neutrophil count (ANC) >500/microL by day 25 +/- 1 and platelets >20,000/microL by day 29 +/ 2. This hematopoietic recovery was delayed in comparison to two animals that received either a graft consisting of freshly isolated, unexpanded CD34(+) cells or 175 x 10(6)/kg unfractionated marrow mononuclear cells. Analysis of the proliferative status of cells in PMVEC expansion cultures demonstrated that by 10 days, 99.8% of CD34(+) cells present in the cultures had undergone cycling, and that the population of cells expressing a CD34(+) CD38(-) phenotype in the cultures was also the result of active cell division. These data indicate that isolated bone marrow CD34(+) cells may undergo cell division during ex vivo expansion in the presence of endothelial cells to provide a graft capable of rescuing a myeloablated autologous host. PMID- 10381504 TI - Erythropoietin- and stem cell factor-induced DNA synthesis in normal human erythroid progenitor cells requires activation of protein kinase Calpha and is strongly inhibited by thrombin. AB - Proliferation, differentiation, and survival of erythroid progenitor cells are mainly regulated by stem cell factor (SCF) and erythropoietin (Epo). Using normal human progenitors, we analyzed the role of Ca2+-sensitive protein kinase C (PKC) subtypes and of G-protein-coupled receptor ligands on growth factor-dependent DNA synthesis. We show that stimulation of DNA synthesis by the two growth factors requires activation of PKCalpha. Inhibitors of Ca2+-activated PKC subtypes blocked the growth factor-induced 3H-thymidine incorporation. SCF and Epo caused no significant translocation of PKCalpha into the membrane, but treatment of intact cells with either of the two cytokines resulted in enhanced activity of immunoprecipitated cytosolic PKCalpha. Stimulation of PKC with the phorbol ester PMA mimicked the cytokine effect on DNA synthesis. Epo-, SCF-, and PMA-induced thymidine incorporation was potently inhibited by thrombin (half-maximal inhibition with 0.1 U/mL). This effect was mediated via the G-protein-coupled thrombin receptor and the Rho guanosine triphosphatase. Adenosine diphosphate caused a modest Ca2+-dependent stimulation of DNA synthesis in the absence of cytokines and specifically enhanced the effect of SCF. Cyclic 3', 5'-adenosine monophosphate exerted a selective inhibitory effect on Epo-stimulated thymidine incorporation. Our results define PKCalpha as major intermediate effector of cytokine signaling and suggest a role for thrombin in controlling erythroid progenitor proliferation. PMID- 10381505 TI - Differentiation of the mononuclear phagocyte system during mouse embryogenesis: the role of transcription factor PU.1. AB - During mouse embryogenesis, macrophage-like cells arise first in the yolk sac and are produced subsequently in the liver. The onset of liver hematopoiesis is associated with the transition from primitive to definitive erythrocyte production. This report addresses the hypothesis that a similar transition in phenotype occurs in myelopoiesis. We have used whole mount in situ hybridization to detect macrophage-specific genes expressed during mouse development. The mouse c-fms mRNA, encoding the receptor for macrophage colony-stimulating factor (CSF 1), was expressed on phagocytic cells in the yolk sac and throughout the embryo before the onset of liver hematopoiesis. Similar cells were detected using the mannose receptor, the complement receptor (CR3), or the Microphthalmia transcription factor (MITF) as mRNA markers. By contrast, other markers including the F4/80 antigen, the macrophage scavenger receptor, the S-100 proteins, S100A8 and S100A9, and the secretory product lysozyme appeared later in development and appeared restricted to only a subset of c-fms-positive cells. Two-color immunolabeling on disaggregated cells confirmed that CR3 and c-fms proteins are expressed on the same cells. Among the genes appearing later in development was the macrophage-restricted transcription factor, PU.1, which has been shown to be required for normal adult myelopoiesis. Mice with null mutations in PU.1 had normal numbers of c-fms-positive phagocytes at 11.5dpc. PU.1(-/-) embryonic stem cells were able to give rise to macrophage-like cells after cultivation in vitro. The results support previous evidence that yolk sac-derived fetal phagocytes are functionally distinct from those arising in the liver and develop via a different pathway. PMID- 10381506 TI - A requirement for K+-channel activity in growth factor-mediated extracellular signal-regulated kinase activation in human myeloblastic leukemia ML-1 cells. AB - Voltage-gated K+ channels have been shown to be required for proliferation of various types of cells. Much evidence indicates that K+-channel activity is required for G1 progression of the cell cycle in different cell backgrounds, suggesting that K+-channel activity is required for early-stage cell proliferation in these cells. However, little is known about the molecular mechanisms that underlie this phenomenon. We have shown in human myeloblastic leukemia ML-1 cells that K+ channels are activated by epidermal growth factor (EGF), whereas serum starvation deprivation suppressed their activity. In addition, voltage-gated K+ channels are required for G1/S-phase transition of the cell cycle. We report here that suppression of K+ channels prevented the activation of extracellular signal-regulated protein kinase 2 (ERK-2) in response to EGF and serum. However, blockade of K+ channels did not prevent ERK-2 activation induced by 12-O-tetradecanoyl-phorbol 13-acetate (TPA). Elimination of extracellular Ca2+ did not alter either ERK-2 activation or the effect of K+ channel blockade on ERK-2 activation. Our data demonstrate that the K+ channel is a part of the EGF-mediated mitogenic signal-transduction process and is required for initiation of the EGF-mediated mitogen-activated protein kinase (MAPK) pathways. Our findings may thus explain why an increase in K+-channel activity is associated with cell proliferation in many types of cells, including ML-1 cells. PMID- 10381507 TI - Abnormal expression and subcellular distribution of subunit proteins of the AP-3 adaptor complex lead to platelet storage pool deficiency in the pearl mouse. AB - The pearl mouse is a model for Hermansky Pudlak Syndrome (HPS), whose symptoms include hypopigmentation, lysosomal abnormalities, and prolonged bleeding due to platelet storage pool deficiency (SPD). The gene for pearl has recently been identified as the beta3A subunit of the AP-3 adaptor complex. The objective of these experiments was to determine if the expression and subcellular distribution of the AP-3 complex were altered in pearl platelets and other tissues. The beta3A subunit was undetectable in all pearl cells and tissues. Also, expression of other subunit proteins of the AP-3 complex was decreased. The subcellular distribution of the remaining AP-3 subunits in platelets, macrophages, and a melanocyte-derived cell line of pearl mice was changed from the normal punctate, probably endosomal, pattern to a diffuse cytoplasmic pattern. Ultrastructural abnormalities in mutant lysosomes were likewise apparent in mutant kidney and a cultured mutant cell line. Genetically distinct mouse HPS models had normal expression of AP-3 subunits. These and related experiments strongly suggest that the AP-3 complex regulates the biogenesis/function of organelles of platelets and other cells and that abrogation of expression of the AP-3 complex leads to platelet SPD. PMID- 10381508 TI - An Arg/Ser substitution in the second epidermal growth factor-like module of factor IX introduces an O-linked carbohydrate and markedly impairs activation by factor XIa and factor VIIa/Tissue factor and catalytic efficiency of factor IXa. AB - Factor IXR94S is a naturally occurring hemophilia B defect, which results from an Arg 94 to Ser mutation in the second epidermal growth factor (EGF)-like module of factor IX. Recombinant factor IXR94S was activated by factor XIa/calcium with an approximately 50-fold reduced rate and by factor VIIa/tissue factor/phospholipid/calcium with an approximately 20-fold reduced rate compared with wild-type factor IX. The apparent molecular mass of the light chain of factor IXaR94S was approximately 6 kD higher than that of plasma or wild-type factor IX, which was not corrected by N-glycosidase F digestion. This result indicated the presence of additional O-linked carbohydrate in the mutant light chain, probably at new Ser 94. The initial rate of activation of factor X by factor IXaR94S in the presence of polylysine was 7% +/- 1% of the initial rate of activation of factor X by plasma factor IXa, and the kc/Km for activation of factor X by factor IXaR94S/factor VIIIa/phospholipid/calcium was 4% +/- 1% of the kc/Km for activation of factor X by plasma factor IXa/factor VIIIa/phospholipid/calcium. The reduced efficiency of activation of factor X by factor IXaR94S in the tenase enzyme complex was due to a 58-fold +/- 12-fold decrease in kcat with little effect on Km. In conclusion, the R94S mutation had introduced an O-linked carbohydrate, which markedly impaired both activation by factor XIa and turnover of factor X in the tenase enzyme complex. PMID- 10381509 TI - Characterization of wild-type and mutant alpha2-antiplasmins: fibrinolysis enhancement by reactive site mutant. AB - During human blood clotting, alpha2-antiplasmin (alpha2AP) becomes covalently linked to fibrin when activated blood clotting factor XIII (FXIIIa) catalyzes the formation of an isopeptide bond between glutamine at position two in alpha2AP and a specific epsilon-lysyl group in each of the alpha-chains of fibrin. This causes fibrin to become resistant to plasmin-mediated lysis. We found that chemically Arg-modified alpha2AP, which lacked plasmin-inhibitory activity, competed effectively with native alpha2AP for becoming cross-linked to fibrin and as a consequence, enhanced fibrinolysis. Recombinant alpha2AP reported to date by other groups either lacked or possessed a low level of FXIIIa substrate activity. As a first step in the development of an engineered protein that might have potential as a localized fibrin-specific fibrinolytic enhancer, we expressed recombinant alpha2AP in Pichia pastoris yeast. Two forms of nonglycosylated recombinant alpha2AP were expressed, isolated and characterized: (1) wild-type, which was analogous to native alpha2AP, and (2) a mutant form, which had Ala substituted for the reactive-site Arg364. Both the wild-type and mutant forms of alpha2AP functioned as FXIIIa substrates with affinities and kinetic efficiencies comparable to those of native alpha2AP, despite each having an additional acetylated Met blocking group at their respective amino-termini. Wild-type recombinant alpha2AP displayed full plasmin inhibitory activity, while mutant alpha2AP had none. Neither the absence of glycosylation nor blockage of the amino terminus affected plasmin-inhibitory or FXIIIa substrate activities of wild-type alpha2AP. When our mutant alpha2AP, which lacked plasmin-inhibitory function, was added to human plasma or whole blood clots, urokinase (UK)-induced clot lysis was enhanced in a dose-dependent manner, indicating that mutant alpha2AP augmented lysis by competing with native alpha2AP for FXIIIa-catalyzed incorporation into fibrin. PMID- 10381510 TI - Contribution of distinct adhesive interactions to platelet aggregation in flowing blood. AB - Aggregation of blood platelets contributes to the arrest of bleeding at sites of vascular injury, but it can occlude atherosclerotic arteries and precipitate diseases such as myocardial infarction. The bonds that link platelets under flow conditions were identified using confocal videomicroscopy in real time. Glycoprotein (GP) Ibalpha and von Willebrand factor (vWF) acted in synergy with alphaIIbbeta3 and fibrinogen to sustain platelet accrual at the apex of thrombi where three-dimensional growth resulted in increasing shear rates. The specific function of distinct adhesion pathways in response to changing hemodynamic conditions helps to explain hemostatic and thrombotic processes. PMID- 10381511 TI - von Willebrand factor propeptide in vascular disorders: A tool to distinguish between acute and chronic endothelial cell perturbation. AB - Before de novo synthesized von Willebrand factor (vWF) leaves the endothelial cell, it undergoes endoproteolytic cleavage of its propeptide (vW antigen II). The processed vWF and propeptide are either released constitutively or, following activation of the endothelium, released through the regulated pathway. In a recent study (Borchiellini et al, Blood 88:2951, 1996), we showed that the half life of mature vWF and of its propeptide differ fourfold to fivefold. We postulated that the molar ratio of the propeptide to mature vWF could serve as a tool to assess the extent of endothelial cell activation under physiologic and clinical conditions. To test this hypothesis, we measured mature vWF and propeptide in patients with documented acute and chronic vascular disease, including patients with thrombotic thrombocytopenic purpura (TTP), acute septicemia, and diabetes mellitus. These data were compared with experimental conditions in healthy subjects in which perturbation of the endothelium was simulated by physical exercise or by administration of 1-deamino-8-D-arginine vasopressin (DDAVP) or endotoxin. In all individuals of the latter study group, both vWF and propeptide levels were elevated during the acute phase of the experimentally induced vascular perturbation; at later time points after stimulation, only vWF levels remained elevated. In patients with sepsis and TTP, both vWF and propeptide were elevated several-fold. Thus, this pattern can readily be explained in terms of acute perturbation of the endothelium. In contrast, in patients with diabetes mellitus propeptide levels were only slightly elevated, whereas vWF levels were elevated twofold to threefold. This pattern is a typical feature of chronic, low-grade activation of the endothelium. These observations support our hypothesis that measurement of both propeptide and vWF levels allows to discriminate between chronic and acute phases of endothelial cell activation in vivo. Measurement of only vWF is less indicative in this respect. PMID- 10381512 TI - Kozak sequence polymorphism of the glycoprotein (GP) Ibalpha gene is a major determinant of the plasma membrane levels of the platelet GP Ib-IX-V complex. AB - Despite the known importance of the sequences surrounding ATG start codons (Kozak sequences) for efficient translation of proteins, few reports have appeared that describe the natural variations in these sequences. Here, we report a human polymorphism in the Kozak sequence of the platelet adhesion receptor, glycoprotein (GP) Ibalpha, a component of the GP Ib-IX-V complex, which mediates the initial adhesion of platelets to the blood vessel wall following injury. The polymorphism is based on the presence of either thymine (T) or cytosine (C) at position -5 from the initiator ATG in the GP Ibalpha gene. The less common allele, -5C, represented 8% to 17% of the alleles in four ethnic populations surveyed. This allele more closely resembles the sequence considered optimal for efficient initiation of protein translation and is associated with increased expression of the receptor on the cell membrane, both in transfected cells and in the platelets of individuals carrying the allele. In vitro transcription/translation studies indicate that the increased expression results from more efficient translation of the -5C form of the GP Ibalpha mRNA. Other mutations made to approximate more closely the consensus sequence described by Kozak did not increase expression of the receptor. This is the first known description of Kozak sequence polymorphism as a determinant of the surface levels of a cell adhesion receptor. This polymorphism may influence an individual's susceptibility for the development of cardiovascular disease. PMID- 10381513 TI - A high-affinity human antibody that targets tumoral blood vessels. AB - Angiogenesis is a characteristic feature of many aggressive tumors and of other relevant disorders. Molecules capable of specifically binding to new-forming blood vessels, but not to mature vessels, could be used as selective vehicles and would, therefore, open diagnostic and therapeutic opportunities. We have studied the distribution of the ED-B oncofetal domain of fibronectin, a marker of angiogenesis, in four different tumor animal models: the F9 murine teratocarcinoma, SKMEL-28 human melanoma, N592 human small cell lung carcinoma, and C51 human colon carcinoma. In all of these experimental models we observed accumulation of the fibronectin isoform containing the ED-B domain around neovascular structures when the tumors were in the exponentially growing phase, but not in the slow-growing phase. Then we performed biodistribution studies in mice bearing a subcutaneously implanted F9 murine teratocarcinoma, using a high affinity human antibody fragment (L19) directed against the ED-B domain of fibronectin. Radiolabeled L19, but not an irrelevant anti-lysozyme antibody fragment (D1.3), efficiently localizes in the tumoral vessels. The maximal dose of L19 accumulated in the tumor was observed 3 hours after injection (8.2% injected dose per gram). By virtue of the rapid clearance of the antibody fragment from the circulation, tumor-to-blood ratios of 1.9, 3.7, and 11.8 were obtained at 3, 5, and 24 hours, respectively. The tumor-targeting performance of L19 was not dose-dependent in the 0.7 to 10 microg range of injected antibody. The integral of the radioactivity localized in tumoral vessels over 24 hours was greater than 70-fold higher than the integral of the radioactivity in blood over the same time period, normalized per gram of tissue or fluid. These findings quantitatively show that new-forming blood vessels can selectively be targeted in vivo using specific antibodies, and suggest that L19 may be of clinical utility for the immunoscintigraphic detection of angiogenesis in patients. PMID- 10381514 TI - Marked temperature dependence of the platelet calcium signal induced by human von Willebrand factor. AB - Interaction of von Willebrand factor (vWF) with the platelet is essential to hemostasis when vascular injury occurs. This interaction elevates the intracellular free calcium concentration ([Ca2+]i) and promotes platelet activation. The present study investigated the temperature dependence of vWF induced [Ca2+]i signaling in human platelets. The influence of temperature can provide invaluable insight into the underlying mechanism. Platelet [Ca2+]i was monitored with Fura-PE3. Ristocetin-mediated binding of vWF induced a transient platelet [Ca2+]i increase at 37 degrees C, but no response at lower temperatures (20 degrees C to 25 degrees C). This temperature dependence could not be attributed to a reduction in vWF binding, as ristocetin-mediated platelet aggregation and agglutination were essentially unaffected by temperature. Most other platelet agonists (U-46619, alpha-thrombin, and adenosine 5'-diphosphate [ADP]) induced a [Ca2+]i signal whose amplitude did not diminish at lower temperatures. The [Ca2+]i signal in response to arachidonic acid, however, showed similar temperature dependence to that seen with vWF. Assessment of thromboxane A2 production showed a strong temperature dependence for metabolism of arachidonic acid by the cyclo-oxygenase pathway. vWF induced thromboxane A2 production in the platelet. Aspirin treatment abolished the vWF-induced [Ca2+]i signal. These observations suggest that release of arachidonic acid and its conversion to thromboxane A2 play a central role in vWF-mediated [Ca2+]i signaling in the platelet at physiological temperatures. PMID- 10381515 TI - Complexes of heparin and platelet factor 4 specifically stimulate T cells from patients with heparin-induced thrombocytopenia/thrombosis. AB - Heparin-induced thrombocytopenia with thrombosis (HITT) is associated with antibodies specific for complexes consisting of heparin and platelet factor 4 (PF4). Studies in individual patients with HITT have demonstrated immunoglobulin (Ig) class switching from IgM to the IgG or IgA isotypes. This transition is thought to require helper T cells, but no studies of the cellular or molecular basis of this process have yet been reported. To characterize T-cell involvement in HITT, peripheral blood mononuclear cells (PBMC) from two patients with classical HITT obtained shortly after the acute episode were restimulated with heparin:PF4 complexes, PF4 alone, heparin alone, and medium alone in the presence of autologous antigen-presenting cells (APC). Responding T cells were then examined using the technique of "spectratyping," in which sequences encoding CDR3 domains of individual V beta (BV) families are amplified and separated by gel electrophoresis. After 14 days in culture with antigen (heparin:PF4 complexes), but not after culture with PF4, heparin, or medium alone, patient cells, but not cells from normal subjects, preferentially expressed T-cell receptor (TCR) containing beta chains of the BV 5.1 family. Nucleotide sequencing of BV 5.1 TCR CDR3 showed that each patient had a personal repertoire, but also shared a tetrapeptide motif (PGTG). These findings provide evidence that the humoral immune response associated with HITT is driven by helper T cells that presumably recognize peptides derived from PF4. Identification of a common beta-chain CDR3 motif in responding T cells from each of two patients suggests that a limited number of helper TCRs may be used to mount an antibody response to heparin:PF4 complexes. TCR spectratyping appears to offer a new way to examine the molecular basis of pathologic immune responses and may be useful in further studies of HITT and other immune-mediated hematologic disorders. PMID- 10381516 TI - Human follicular dendritic cells inhibit superantigen-induced T-cell proliferation by distinct mechanisms. AB - Follicular dendritic cells (FDCs) reside within germinal centers of secondary lymphoid tissue where they play a critical role in antigen-driven immune responses. FDCs express numerous adhesion molecules that facilitate cellular interactions with B and T cells within the germinal center microenvironment. Although human FDCs have been shown to influence B-cell development, very little is known about the ability of FDCs to regulate T-cell responses. To investigate this functional aspect of FDCs, highly enriched preparations were isolated by magnetic cell separation using the FDC-restricted monoclonal antibody HJ2. We found that isolated human FDCs inhibited proliferation of both autologous and allogeneic T cells, and were dependent on the number of FDCs present. Inhibition by FDCs was observed using two serologically distinct superantigens at multiple concentrations (Staphylococcus enterotoxin A and B). In contrast, B cells failed to inhibit, and often augmented superantigen-induced T-cell proliferation. Antibody-blocking studies showed that CD54 and CD106 were involved in the ability of FDC to inhibit T-cell proliferative responses. When FDCs and T cells were separated by a semipermeable membrane, the inhibitory effect was partially abrogated, demonstrating that in addition to cell-cell interactions, a soluble factor(s) was also involved in the process. The addition of indomethicin to cultures improved the proliferative response in the presence of FDCs, indicating that inhibition was mediated, in part, by prostaglandins. These results indicate that FDCs regulate T-cell proliferation by two molecular mechanisms and that FDC:T-cell interactions may play a pivotal role in germinal center development. PMID- 10381517 TI - Isochromosome 17q in blast crisis of chronic myeloid leukemia and in other hematologic malignancies is the result of clustered breakpoints in 17p11 and is not associated with coding TP53 mutations. AB - An isochromosome of the long arm of chromosome 17, i(17q), is the most frequent genetic abnormality observed during the disease progression of Philadelphia chromosome-positive chronic myeloid leukemia (CML), and has been described as the sole anomaly in various other hematologic malignancies. The i(17q) hence plays a presumably important pathogenetic role both in leukemia development and progression. This notwithstanding, the molecular consequences of this abnormality have not been investigated in detail. We have analyzed 21 hematologic malignancies (8 CML in blast crisis, 8 myelodysplastic syndromes [MDS], 2 acute myeloid leukemias, 2 chronic lymphocytic leukemias, and 1 acute lymphoblastic leukemia) with i(17q) by fluorescence in situ hybridization (FISH). Using a yeast artificial chromosome (YAC) contig, derived from the short arm of chromosome 17, all cases were shown to have a breakpoint in 17p. In 12 cases, the breaks occurred within the Smith-Magenis Syndrome (SMS) common deletion region in 17p11, a gene-rich region which is genetically unstable. In 10 of these 12 cases, we were able to further map the breakpoints to specific markers localized within a single YAC clone. Six other cases showed breakpoints located proximally to the SMS common deletion region, but still within 17p11, and yet another case had a breakpoint distal to this region. Furthermore, using chromosome 17 centromere specific probes, it could be shown that the majority of the i(17q) chromosomes (11 of 15 investigated cases) were dicentric, ie, they contained two centromeres, strongly suggesting that i(17q) is formed through an intrachromosomal recombination event, and also implicating that the i(17q), in a formal sense, should be designated idic(17)(p11). Because i(17q) formation results in loss of 17p material, potentially uncovering the effect of a tumor suppressor on the remaining 17p, the occurrence of TP53 mutations was studied in 17 cases by sequencing the entire coding region. In 16 cases, no TP53 mutations were found, whereas one MDS displayed a homozygous deletion of TP53. Thus, our data suggest that there is no association between i(17q) and coding TP53 mutations, and that another tumor suppressor gene(s), located in proximity of the SMS common deletion region, or in a more distal location, is of pathogenetic importance in i(17q) associated leukemia. PMID- 10381518 TI - The importance of antibody-specificity in determining successful radioimmunotherapy of B-cell lymphoma. AB - We report the radioimmunotherapy of mouse B-cell lymphoma, BCL1, using a panel of anti-B-cell monoclonal antibodies (MoAb) (anti-CD19, anti-CD22, anti-major histocompatibility complex (MHC) II, and anti-idiotype (Id) radiolabeled with 131 iodine. When administered early in disease (day 4), the 131I-anti-MHCII MoAb cured tumors as a result of targeted irradiation alone, the unlabeled MoAb being nontherapeutic. In contrast, 131I-anti-Id, despite targeting irradiation and having therapeutic activity as an unconjugated antibody, protected mice for only 30 days; 131I-anti-CD19 and anti-CD22 were therapeutically inactive. Binding and biodistribution studies showed that the anti-Id, unlike anti-MHCII, MoAb was cleared from target cells in vivo and delivered 4 times less irradiation to splenic tumor. Treating later in the disease (day 14) increased tumor load and produced the expected reduction in therapeutic activity with the anti-MHCII, but surprisingly, allowed 131I-anti-Id to cure most mice. This unexpected potency of 131I-anti-Id late in the disease appeared to result from the direct cytotoxicity of the anti-Id MoAb, which was more active in established disease, in combination with targeted irradiation. We believe the ability of targeted irradiation and certain cytotoxic MoAb to work cooperatively against tumor in this way has important implications for the selection of reagents in radioimmunotherapy of B cell lymphoma. PMID- 10381519 TI - EBNA-1 gene sequences in Brazilian and American patients with Hodgkin's disease. AB - We examined the types of Epstein-Barr virus-associated nuclear antigen-1 (EBNA-1) gene carboxy (C)-terminal mutations occurring in Hodgkin's disease (HD) and reactive tissues from two different geographic regions. Previously reported EBNA 1 C-terminal region amino acid sequence variants, based on the amino acid at codon 487, include Prototype (P)-ala, which is found in the B95.8-derived prototype virus, P-thr, Variant (V)-leu, V-val, and V-pro. Using polymerase chain reaction (PCR) to amplify portions of the EBNA-1 gene, followed by DNA sequencing, we found a single EBNA-1 gene sequence variant in each tissue, whether reactive or neoplastic and whether from Brazil or the United States. Variant EBNA-1 gene sequences were more common in both neoplastic and non neoplastic tissues from different geographic areas than the so-called prototype sequence. In the 17 Brazilian HD cases, 4 cases had P-thr variants and 13 had V leu variants. In the six reactive tissues from Brazil, one had a P-ala variant, two had P-thr variants, and three had V-leu variants. In the 12 American HD cases, 2 had P-ala variants, 6 had P-thr variants, and 4 had V-leu variants. The 11 American reactive tissues included 2 P-ala variants, 5 P-thr variants, and 4 V leu variants. In both countries, there were similar variant EBNA-1 sequences present in normal tissues and HD cases. Compared with the P-ala and P-thr cases, the V-leu cases were more likely to have the 30-bp latent membrane protein 1 (LMP1) gene deletion (P = 0.0075). In addition, cases of HD with the V-leu were statistically associated with a substitution of asparagine for glutamine at codon 322 of the C-terminal portion of the LMP1 gene. Our results suggest that any variation in EBNA-1 gene sequence is caused by a polymorphism present in pre existing viral strains in the underlying population, and not a mutation occurring during oncogenesis. PMID- 10381520 TI - Growth inhibition of a human myeloma cell line by all-trans retinoic acid is not mediated through downregulation of interleukin-6 receptors but through upregulation of p21(WAF1). AB - All-trans retinoic acid (ATRA) has previously been shown to inhibit the growth of OPM-2 human myeloma cells. The growth inhibition was postulated to result from a transcriptional downregulation of interleukin-6 receptor alpha (IL-6Ralpha) with IL-6Rbeta (gp130) unaffected. To formally test this hypothesis, an expression vector designed for constitutive IL-6Ralpha expression was constructed and used for transfection of OPM-2 cells. Six stable transfectants were cloned. The expression of IL-6Ralpha was shown by immunofluorescence with anti-IL-6Ralpha antibody and 125I-IL-6 binding. In five of six transfectant clones, cellular IL 6Ralpha was 1.5- to 6-fold higher than the parental cells, with the ligand binding affinity unchanged. While ATRA reduced IL-6Ralpha expression in the parental OPM-2 cells, it enhanced its expression in these five transfectants. The clonogenic growth of these transfectants, however, remained strongly inhibited by ATRA. Further analysis, comparing the parental OPM-2 cells and a representative transfectant, clone C5, showed that IL-6 caused rapid tyrosine phosphorylation of gp130 in both OPM-2 and C5 clones. Pretreatment with ATRA greatly reduced IL-6 induced gp130 phosphorylation in OPM-2 cells, reflecting a reduction in cellular IL-6Ralpha. In contrast, IL-6-induced gp130 phosphorylation was not reduced by ATRA pretreatment in C5 cells, indicating that the expressed IL-6Ralpha was functional. Similar to OPM-2 cells, C5 cells were sensitive to growth inhibition by dexamethasone, which was entirely reversed by exogenous IL-6, suggesting that the IL-6 postreceptor signal transduction remained intact. ATRA was further shown to upregulate p21(WAF1) expression and cause dephosphorylation of the retinoblastoma protein (pRB) in both OPM-2 and C5 cells. Exogenous IL-6 also failed to reverse these effects of ATRA. Thus, the growth inhibitory activity of ATRA is not mediated through cellular IL-6Ralpha downregulation and is likely to result from a direct upregulation of p21(WAF1) and consequent dephosphorylation of pRB. PMID- 10381521 TI - Mucosal intra-epithelial lymphocytes in enteropathy-associated T-cell lymphoma, ulcerative jejunitis, and refractory celiac disease constitute a neoplastic population. AB - Loss of response to a gluten-free diet (refractory sprue) and ulcerative jejunitis are complications of celiac disease that may progress to enteropathy associated T-cell lymphoma (EATL). Both conditions are characterized by the presence of a nonlymphomatous monoclonal T-cell population in the enteropathic mucosa. In EATL, a similar monoclonal population that shows clonal identity with the lymphoma itself is also present in the enteropathic mucosa. In this study we show that in all three circumstances the monoclonal T-cell population is constituted by cytologically normal, noninvasive intraepithelial T lymphocytes that share an identical aberrant immunophenotype with EATL. Patients with refractory sprue and/or ulcerative jejunitis are, therefore, suffering from a neoplastic T-cell disorder for which hematological treatment strategies need to be devised. PMID- 10381522 TI - Myeloma cells selected for resistance to CD95-mediated apoptosis are not cross resistant to cytotoxic drugs: evidence for independent mechanisms of caspase activation. AB - We have previously shown that selection for resistance to the anthracenes, doxorubicin or mitoxantrone, results in coselection for resistance to CD95 mediated apoptosis (Landowski et al: Blood 89:1854, 1997). In the present study, we were interested in determining if the converse is also true; that is, does selection for CD95 resistance coselect for resistance to chemotherapeutic drugs. To address this question, we used two isogenic models of CD95-resistant versus CD95-sensitive cell lines: 8226/S myeloma cells selected for resistance to CD95 mediated apoptosis; and K562 cells expressing ectopic CD95. Repeated exposure of the CD95-sensitive human myeloma cell line, 8226/S, to agonistic anti-CD95 antibody resulted in a cell line devoid of CD95 receptor surface expression and completely resistant to CD95-mediated apoptosis. Multiple clonal populations derived from the CD95-resistant cell line showed no difference in sensitivity to doxorubicin, mitoxantrone, Ara-C, or etoposide, demonstrating that cross resistance between Fas-mediated apoptosis and drug-induced apoptosis occurs only when cytotoxic drugs are used as the selecting agent. Using the inverse approach, we transfected the CD95-negative cell line, K562, with a CD95 expression vector. Clones expressing variable levels of cell-surface CD95 were isolated by limiting dilution, and analyzed for sensitivity to CD95-mediated apoptosis and response to chemotherapeutic drugs. We show that CD95 surface expression confers sensitivity to CD95-mediated apoptosis; however, it does not alter response to chemotherapeutic drugs. Similarly, doxorubicin-induced activation of caspases 3 and 8 was identical in the CD95-sensitive and CD95-resistant cell lines in both isogenic cell systems. In addition, prior treatment with the CD95 receptor blocking antibody, ZB4, inhibited CD95-activated apoptosis in 8226/S cells, but had no effect on doxorubicin cytotoxicity. These results show that CD95 and chemotherapeutic drugs use common apoptotic effectors, but the point of convergence in these two pathways is downstream of CD95 receptor/ligand interaction. PMID- 10381523 TI - Expression of the human immunodeficiency virus-Tat gene in lymphoid tissues of transgenic mice is associated with B-cell lymphoma. AB - The human immunodeficiency virus type 1 (HIV-1) Tat gene, a potent transactivator of viral and cellular genes, has been proposed as a key agent in the pathogenesis of acquired immune deficiency syndrome related disorders, including non-Hodgkin's lymphoma. In cultured cells, the HIV-1 Tat protein can induce the expression of the cytokines interleukin-6 (IL-6) and IL-10, which are known to induce proliferation and differentiation of lymphoid cells. Such alterations in cytokine expression, together with a secondary genetic event, are thought to ultimately lead to oncogenic transformation. To address the influence of Tat on lymphoid development in the context of the whole organism, we produced several transgenic mouse lines that express the Tat gene under the control of an actin promoter. We show here that this promoter directs expression to a variety of sites, including spleen, bone marrow, and lymph nodes. Approximately 25% to 30% of the Tat transgenic population developed enlarged spleens within 1 year after birth. On histological examination, a significant number of spleens from Tat-transgenic mice exhibited malignant lymphoma of B-cell origin. IgG heavy chain rearrangement confirmed the clonal B-cell nature of these lymphoproliferations. In contrast, T cell receptor genes exhibited a germline (unrearranged) structure. Reverse transcription polymerase chain reaction analysis of transgenic spleens revealed that mRNA encoding cytokines IL-6 and IL-10 was upregulated, suggesting a possible mechanism for the B-cell expansion in vivo. PMID- 10381524 TI - Loss of heterozygosity and microsatellite instability at the MLL locus are common in childhood acute leukemia, but not in infant acute leukemia. AB - Rearrangements involving the MLL gene at chromosome 11q23 are associated with leukemia and are present in up to 70% of infant leukemias. Loss of heterozygosity (LOH) has been shown for anonymous polymorphic markers at 11q23 in adult leukemias. To study LOH at the MLL locus, we have identified two new polymorphic microsatellite markers: a GAA repeat (mllGAAn) in intron 6 of the MLL gene and a GA (mllGAn) repeat in the 5' flanking region of the gene, approximately 2 kb upstream of the translation initiation codon. The heterozygosity index of mllGAAn is 0.54, which renders it useful for analyzing LOH. We screened two groups of leukemia patients to study LOH at the mllGAAn marker. Group A (n = 18) was selected on the basis of presentation before 18 months. Cytogenetic and reverse transcription-polymerase chain reaction analysis showed that 9 of these 18 children had translocations involving MLL. No LOH was observed. Group B (n = 36) were randomly selected children who had presented with leukemia between 1993 and 1994. Cytogenetic analysis of this group showed a variety of different chromosomal abnormalities. LOH was shown in 9 of 20 individuals (45%) who were informative. Microsatellite instability (MSI) was demonstrated in 1 of 18 individuals in group A and 5 of 36 individuals (13.9%) in group B. MSI and LOH were observed simultaneously in three individuals. Loss of an allele was confirmed in one individual by fluorescence in situ hybridization. Individuals with MSI or LOH at mllGAAn were selected for analysis at anonymous polymorphic markers D11S1364 and D11S1356, which flank the MLL gene. No LOH or MSI was observed at these markers in those individuals who were informative. These results show that LOH at the MLL gene locus is a common event during leukemogenesis. Furthermore, the presence of MSI at this locus suggests that the region is a hotspot for genetic instability. PMID- 10381525 TI - Caspase-mediated proteolysis and activation of protein kinase Cdelta plays a central role in neutrophil apoptosis. AB - Neutrophils undergo constitutive apoptosis when aged ex vivo. Recent studies have indicated roles for Fas/CD95 and the nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase system in this process. We have investigated the role of protein kinase C (PKC) in neutrophil death. We show that there is proteolysis and activation of the novel isoform PKCdelta in aged neutrophils and that this process is accelerated by the addition of an agonistic Fas antibody. PKCdelta proteolysis occurs before the onset of any detectable features of apoptosis and pharmacologic inhibition of this enzyme inhibits neutrophil apoptosis. PKCdelta cleavage and activation is dependent on caspase-8/FADD-like interleukin-1beta converting enzyme (FLICE)-mediated processing of caspase-3/CPP32. Neutrophil survival is prolonged by the addition of broad spectrum (BD.fmk) or caspase-8 targeted (zIETD.fmk) peptide caspase inhibitors. Inhibition of PKCdelta does not prevent apoptosis triggered by factor withdrawal in immature hematopoietic cells, including normal human CD34(+) progenitors indicating that within a given lineage, the mechanisms of apoptosis may be differentiation-stage-specific. Ex vivo aging of neutrophils leads to the increasing production of reactive oxygen species and this is attenuated in cells treated with either caspase or PKCdelta inhibitors. Proteolytically activated PKCdelta acts as a molecular link between the Fas/CD95 receptor and the NADPH-oxidase system and plays a central role in regulating the process of neutrophil apoptosis. PMID- 10381526 TI - The carboxy-terminal cell-binding domain of thrombospondin is essential for sickle red blood cell adhesion. AB - Sickle red blood cells (SS-RBCs) have enhanced adhesion to the plasma and subendothelial matrix protein thrombospondin-1 (TSP) under conditions of flow in vitro. TSP has at least four domains that mediate cell adhesion. The goal of this study was to map the site(s) on TSP that binds SS-RBCs. Purified TSP proteolytic fragments containing either the N-terminal heparin-binding domain, or the type 1, 2, or 3 repeats, failed to sustain SS-RBC adhesion (<10% adhesion). However, a 140-kD thermolysin TSP fragment, containing the carboxy-terminal cell-binding domain in addition to the type 1, 2, and 3 repeats fully supported the adhesion of SS-RBCs (126% +/- 25% adhesion). Two cell-binding domain adhesive peptides, 4N1K (KRFYVVMWKK) and 7N3 (FIRVVMYEGKK), failed to either inhibit or support SS RBC adhesion to TSP. In addition, monoclonal antibody C6. 7, which blocks platelet and melanoma cell adhesion to the cell-binding domain, did not inhibit SS-RBC adhesion to TSP. These data suggest that a novel adhesive site within the cell binding domain of TSP promotes the adhesion of sickle RBCs to TSP. Furthermore, soluble TSP did not bind SS-RBCs as detected by flow cytometry, nor inhibit SS-RBC adhesion to immobilized TSP under conditions of flow, indicating that the adhesive site on TSP that recognizes SS-RBCs is exposed only after TSP binds to a matrix. We conclude that the intact carboxy-terminal cell-binding domain of TSP is essential for the adhesion of sickle RBCs under flow conditions. This study also provides evidence for a unique adhesive site within the cell binding domain that is exposed after TSP binds to a matrix. PMID- 10381527 TI - Identification of a defect in the intracellular trafficking of a Kell blood group variant. AB - Blood group polymorphisms have been used as tools to study the architecture of the red blood cell (RBC) membrane. Some blood group variants have reduced antigen expression at the cell surface. Understanding the underlying mechanism for this reduced expression can potentially provide structural information and help to elucidate protein trafficking pathways of membrane proteins. The Kp(a+) phenotype is a variant in the Kell blood group system that is associated with a single amino acid substitution (R281W) in the Kell glycoprotein and serologically associated with a weakened expression of other Kell system antigens by an unknown mechanism. We found by immunoblotting of RBCs that the weakening of Kell antigens in this variant is due to a reduced amount of total Kell glycoprotein at the cell surface rather than to the inaccessibility of the antigens to Kell antibodies. Using a heterologous expression system, we demonstrate that the Kpa mutation causes retention of most of the Kell glycoprotein in a pre-Golgi compartment due to differential processing, thereby suggesting aberrant transport of the Kell protein to the cell surface. Furthermore, we demonstrated that single nucleotide substitutions into the coding region of the common KEL allele, as predicted by the molecular genotyping studies, was sufficient to encode three clinically significant low incidence antigens. We found that two low incidence antigens can be expressed on a single Kell protein, thus showing that the historical failure to detect such a variant is not due to structural constraints in the Kell protein. These studies demonstrate the power of studying the molecular mechanisms of blood group variants for elucidating the intracellular transport pathways of membrane proteins and the requirements for cell surface expression. PMID- 10381528 TI - Hereditary spherocytosis and elliptocytosis erythrocytes show a normal transbilayer phospholipid distribution. AB - Phosphatidylserine (PS) asymmetry was determined in red blood cells from patients with hereditary spherocytosis and elliptocytosis. No PS-exposing subpopulations were detected using the very sensitive method with fluorescently labeled annexin V. Treatment with N-ethylmaleimide or adenosine triphosphate (ATP) depletion to inactivate the flipase did not lead to formation of PS-exposing subpopulations in these cells, but elevated intracellular calcium levels did lead to extensive scrambling of the PS asymmetry. Although interactions of the membrane skeleton with the phospholipid bilayer have been suggested to stabilize the asymmetric distribution of PS across the bilayer, our data show that red blood cells with a severely damaged membrane skeleton are able to preserve asymmetry, even under conditions in which restoration of the asymmetric distribution is excluded. Moreover, the loss of membrane asymmetry in these cells requires active scrambling involving high levels of intracellular calcium as in normal cells. Our data show that the severe disorder of the membrane skeleton found in these cells does not affect the activity of flipase or scramblase, indicating that these proteins are not regulated by, nor coupled to the membrane skeleton assembly, and that possible thrombotic events in spherocytosis patients are not likely associated with altered PS topology of the red blood cells. PMID- 10381529 TI - Inhibitory activity of human lactoferrin and its peptide on chondroitin sulfate A , CD36-, and thrombospondin-mediated cytoadherence of plasmodium falciparum infected erythrocytes. AB - Lactoferrin (LF), a human serum protein, strongly inhibited the adherence of Plasmodium falciparum-infected erythrocytes (PE) to immobilized chondroitin sulfate A (CSA)-conjugated albumin at a concentration of 100 microg/mL and blocked the PE binding to CD36-expressing Chinese hamster ovary (CHO) cells, as well as immobilized CD36 at concentrations of 5 microg/mL and 100 microg/mL, respectively. Biotinylated LF bound to CD36 in a saturable manner, and such binding was inhibited by unlabeled LF and the anti-CD36 monoclonal antibody, 8A6, suggesting specificity of binding. Additionally, LF inhibited PE binding to immobilized thrombospondin (TSP) at a concentration of 100 microg/mL, and specific binding of LF to TSP was confirmed using biotinylated LF. LF inhibited PE binding to C32 amelanotic melanoma cells in a dose-dependent manner. A peptide of LF, Arg-Asn-Met Arg-Lys-Val Arg-Gly-Pro-Pro-Val-Ser-Cys (amino acid residues 25-37 of LF), which has been suggested to contribute to LF binding to various materials, including CSA, inhibited PE binding to immobilized CSA-conjugated albumin, immobilized CD36, CD36-expressing CHO cells, immobilized TSP, and C32 amelanotic melanoma cells, as well as LF itself. These results suggest that LF peptide may provide the basis for developing agents that are able to inhibit CSA , CD36-, and TSP-mediated cytoadherence of PE. PMID- 10381530 TI - Role of natural killer cell alloreactivity in HLA-mismatched hematopoietic stem cell transplantation. AB - Because of the expression of inhibitory receptors (KIR) for major histocompatibility complex (MHC) class I allotypes, a person's natural killer (NK) cells will not recognize and will, therefore, kill cells from individuals lacking his/her KIR epitopes. This study investigated the role of NK cell alloreactivity in human HLA haplotype-mismatched hematopoietic stem cell transplantation and, specifically, the role of the three major NK specificities, ie, those for HLA-C group 1, HLA-C group 2, and HLA-Bw4 alleles. In 20 of 60 donor-recipient pairs, KIR epitope incompatibility and functional analyses of donor NK cell clones predicted donor NK cells could cause graft-versus-host (GVH)/graft-versus-leukemia (GVL) reactions. NK cell clones of donor origin were obtained from transplanted recipients and tested for lysis of recipient's cryopreserved pretransplant lymphocytes. Despite the absence of GVH disease, we detected high frequencies of NK clones which killed recipient's target cells. Lysis followed the rules of NK cell alloreactivity, being blocked only by the MHC class I KIR epitope which was missing in the recipient. The alloreactive NK clones also killed the allogeneic leukemia. Transplants from these KIR epitope incompatible donors had higher engraftment rates. Therefore, a GVL effector and engraftment facilitating mechanism, which is independent of T-cell-mediated GVH reactions, may be operational in HLA mismatched hematopoietic cell transplants. PMID- 10381531 TI - Incidence of tumor-cell contamination in leukapheresis products of breast cancer patients mobilized with stem cell factor and granulocyte colony-stimulating factor (G-CSF) or with G-CSF alone. AB - We have assessed tumor contamination of peripheral blood progenitor cells (PBPC) in 203 high-risk breast cancer patients who were prospectively randomized to mobilization with stem cell factor (SCF) plus granulocyte colony-stimulating factor (G-CSF) versus G-CSF alone. The patients then received high-dose cyclophosphamide, cisplatin, and carmustine (BCNU) with PBPC support. One bone marrow aspirate obtained before treatment, one whole blood specimen obtained before cytokine infusion, and one to five leukapheresis products were tested for the presence of tumor cells by an alkaline phosphatase immunocytochemical technique with a targeted sensitivity of 1.7 tumor cells per 10(6) hematopoietic cells. Tumor cells were detected in the bone marrow, peripheral blood, and/or PBPC of 21 patients (10%). In 14 patients, bone marrow specimens were tumor positive; in seven patients, premobilization whole blood specimens were tumor positive, and in eight patients, leukapheresis products were tumor-positive. In five patients, repetitive or multiple specimens were tumor-positive, and in three cases, marrow, peripheral blood, and PBPC products were all tumor-positive. Nine of the patients in whom tumor cells were found in marrow or peripheral blood were clinical stage II to III and 12 were clinical stage IV. Nine of the tumor positive patients were in the SCF + G-CSF arm and 12 were in the G-CSF arm. Tumor cells were detected in leukapheresis products of eight patients: three in the G CSF + SCF arm and five in the G-CSF arm. We conclude that detectable tumor-cell contamination of bone marrow, peripheral blood, and/or PBPC occurred in approximately 10% of patients in this trial and was observed in stage II to III patients, as well as in stage IV patients. No significant difference could be found in the rate of PBPC tumor-cell contamination between patients who received SCF + G-CSF compared with those who received G-CSF alone. Neither mobilization regimen was found to increase the rate of tumor-cell contamination when control premobilization blood samples were compared with leukapheresis products. PMID- 10381532 TI - Incomplete T-cell immune reconstitution in two major histocompatibility complex class II-deficiency/bare lymphocyte syndrome patients after HLA-identical sibling bone marrow transplantation. AB - To study the effects of major histocompatibility complex (MHC) class II expression on T-cell development, we have investigated T-cell immune reconstitution in two MHC class II-deficiency patients after allogeneic bone marrow transplantation (allo-BMT). Our study showed that the induction of MHC class II antigen expression on BM graft-derived T cells in these allo-BMT recipients was hampered upon T-cell activation. This reduction was most striking in the CD8(+) T-cell subset. Furthermore, the peripheral T-cell receptor (TCR) repertoire in these graft-derived MHC class II-expressing CD4(+) and in the CD8(+) T-cell fractions was found to be restricted on the basis of TCR complementarity determining region 3 (CDR3) size profiles. Interestingly, the T cell immune response to tetanus toxoid (TT) was found to be comparable to that of the donor. However, when comparing recipient-derived TT-specific T cells with donor-derived T cells, differences were observed in TCR gene segment usage and in the hydropathicity index of the CDR3 regions. Together, these results reveal the impact of an environment lacking endogenous MHC class II on the development of the T-cell immune repertoire after allo-BMT. PMID- 10381534 TI - t(1;2)(q21;p23) and t(2;3)(p23;q21): two novel variant translocations of the t(2;5)(p23;q35) in anaplastic large cell lymphoma. AB - Cytogenetic investigations in two cases of anaplastic large cell lymphoma (ALCL) showed novel variants of the classical (2;5)(p23;q35) translocation, namely a t(1;2)(q21;p23) and a t(2;3)(p23;q21). The tumor cells in both cases gave positive immunohistochemical labeling for ALK protein (with both monoclonal and polyclonal antibodies), demonstrating that these translocations induce aberrant expression of this kinase and suggesting that genes other than NPM can activate the ALK gene in ALCL. These two cases were shown by an in vitro kinase assay to express ALK kinases (104 kD and 97 kD, respectively), which differed in size from the classical NPM-ALK fusion product (80 kD). Moreover, ALK expression was confined to the cytoplasm of the tumor cells in each case, supporting the hypothesis that the observed nuclear localization of NPM-ALK in classical ALCL is not the site of oncogenic activity of the ALK kinase. PMID- 10381533 TI - Embryonic hemoglobins are expressed in definitive cells. AB - Human embryonic zeta and epsilon globin chains are synthesized in yolk sac derived primitive erythroid cells, and decrease rapidly during definitive erythropoiesis. Examination of zeta and epsilon globin expression at the cellular level using dual-color immunofluorescence staining with specific monoclonal antibodies showed that embryonic globin proteins are present in definitive erythroid cells. More than half of fetal erythrocytes were positive for zeta and approximately 5% for epsilon globin. Approximately one third of newborn red blood cells were zeta-positive and less than 1% epsilon-positive. Adult erythrocytes did not have embryonic globins. Erythroblasts that developed in liquid cultures also contained embryonic globin in amounts which declined with ontogenic age, and the proportion of positive cells in vitro was less than in the comparable erythrocytes that developed in vivo. Thus, embryonic globin chains are synthesized in definitive erythroid cells and decrease with ontogeny. Modulation of embryonic globin gene expression is not solely due to a switch from primitive to definitive erythropoiesis. PMID- 10381535 TI - Deletion of the extracellular membrane-distal cytokine receptor homology module of Mpl results in constitutive cell growth and loss of thrombopoietin binding. AB - The thrombopoietin receptor, Mpl, is a member of the cytokine receptor superfamily. The extracellular domain of Mpl contains two copies of the cytokine receptor homology module (CRM). Mpl is encoded by c-mpl, the cellular homologue of the oncogene v-mpl. The oncogenic potential of v-mpl may arise from deletion of all but the 43 most membrane-proximal amino acids of the extracellular domain of the wild-type receptor. To test the hypothesis that the extracellular domain of Mpl plays a role in controlling receptor activity, we created mutants of murine Mpl in which the membrane-distal CRM was either deleted or replaced by the membrane-proximal CRM. Introduction of these mutant receptors into factor dependent BaF3 cells led to constitutive cell growth in the absence of growth factor. Both mutant receptors failed to bind 125I-Tpo. These results suggest that the membrane-distal CRM of Mpl acts as a brake on cell proliferation and that this region is required for ligand binding. PMID- 10381536 TI - A clinical trial of retroviral-mediated transfer of a rev-responsive element decoy gene into CD34(+) cells from the bone marrow of human immunodeficiency virus-1-infected children. AB - Genetic modification of hematopoietic stem cells with genes that inhibit replication of human immunodeficiency virus-1 (HIV-1) could lead to development of T lymphocytes and monocytic cells resistant to HIV-1 infection after transplantation. We performed a clinical trial to evaluate the safety and feasibility of this procedure, using bone marrow from four HIV-1-infected pediatric subjects (ages 8 to 17 years). We obtained bone marrow, isolated CD34(+) cells, performed in vitro transduction with a retroviral vector carrying a rev-responsive element (RRE) decoy gene, and reinfused the cells into these subjects with no evidence of adverse effects. The levels of gene-containing leukocytes in peripheral blood samples in the 1 year after gene transfer/cell infusion have been extremely low. These observations support the potential of performing gene therapy for HIV-1 using hematopoietic cells, but emphasize the need for improved gene transfer techniques. PMID- 10381537 TI - Changes in the gene expression of GABAA receptor subunit mRNAs in the septum of rats subjected to pentylenetetrazol-induced kindling. AB - Chemical kindling was induced in rats by long-term administration of pentylenetetrazol (PTZ) (30 mg/kg three times a week for 9 weeks). The effects of such kindling on the abundance of transcripts encoding subunits of the gamma aminobutyric acid type A (GABAA) receptor in the brain were measured by RNase protection assay. Kindled rats were examined either 3 or 30 days after discontinuation of PTZ treatment. The amounts of gamma2L and gamma2S subunit mRNAs were significantly increased in the hippocampus and cerebral cortex of kindled rats 3 and 30 days after treatment discontinuation, compared with those observed in control rats, and these effects were prevented by the concomitant administration of the anticonvulsant abecarnil. In contrast, the amounts of alpha1 and beta2 subunit mRNAs in these two brain regions did not differ significantly between kindled and control rats. The abundance of alpha1, beta2, gamma2L and gamma2S subunit mRNAs was decreased in the septum of rats 3 or 30 days after discontinuation of treatment with PTZ either alone or in combination with abecarnil. The amounts of none of the four subunit mRNAs measured differed significantly between the striatum or frontal cortex of kindled rats and control rats 3 days after drug discontinuation. Immunohistochemical analysis with antibodies to choline acetyltransferase revealed a marked decrease in the number of cholinergic neurons in the septum of kindled rats 30 days after discontinuation of PTZ treatment; this effect was not prevented by the administration of abecarnil. These results suggest that long-term treatment with PTZ induces a loss of GABAA receptors in the septum. PMID- 10381538 TI - Differential expression of the small GTP-binding proteins RhoA, RhoB, Cdc42u and Cdc42b in developing rat neocortex. AB - Studies with cultured cells indicate that small GTPases of the Rho family may be involved in cell proliferation, differentiation, as well as migration. Therefore, we have studied the expression of four members of this protein family, i.e., RhoA, RhoB, the ubiquitous Cdc42u, and brain specific Cdc42b, during the embryonic and early postnatal development of rat neocortex. A clear isoform specificity of expression was found during the prenatal development. Thus, RhoA and Cdc42u were present in the proliferation zone while RhoB and Cdc42b were expressed only in the cortical plate where neural cells settle and differentiate. After birth, this isoform specificity quickly disappeared so that already at postnatal day 8 the adult pattern of expression was present. Our findings of a differential expression of the small GTP-binding proteins RhoA, RhoB, Cdc42u and Cdc42b in developing brain neocortex suggest isoform specific functions during neurogenesis and differentiation. PMID- 10381539 TI - Brain-derived neurotrophic factor enhances association of protein tyrosine phosphatase PTP1D with the NMDA receptor subunit NR2B in the cortical postsynaptic density. AB - Our recent studies revealed that brain-derived neurotrophic factor (BDNF) rapidly enhances tyrosine phosphorylation and dephosphorylation of the NMDA receptor subunit, NR2B, in the postsynaptic density (PSD), potentially regulating synaptic plasticity. To explore the molecular mechanisms underlying synaptic NR2B signaling, we examined the protein tyrosine phosphatase, PTP1D; BDNF reportedly increases association of PTP1D with tyrosine phosphorylated proteins in cortical neurons and PC 12 cells. We now report that PTP1D is an intrinsic component of the rat cerebrocortical PSD, based on Western blot analysis using specific anti PTP1D antibodies. In addition, NR2B was co-immunoprecipitated with PTP1D using anti-NR2B antibodies or anti-PTP1D antibodies, indicating physical association of the subunit with PTP1D. Moreover, treatment of the purified PSD with BDNF for 5 min elicited a two-fold increase in the association of NR2B with PTP1D. The BDNF action appeared to be specific, since nerve growth factor, another member of the neurotrophin gene family, did not alter the association. Finally, an overlay assay revealed that BDNF caused a two-fold increase in binding of blotted PSD NR2B proteins to PTP1D-SH2 domains, revealing molecular mechanisms mediating the PTP1D-NR2B binding. Taken together, our results raise the possibility that PTP1D participates in BDNF-mediated NR2B signaling cascades at the postsynaptic site, thereby regulating synaptic plasticity. PMID- 10381540 TI - Modulation of apolipoprotein D and apolipoprotein E expression in rat hippocampus after entorhinal cortex lesion. AB - Apolipoprotein (apo) D is a member of the lipocalin family of proteins. Although its physiological function is unknown, apoD is thought to transport one or more small hydrophobic ligands. A second apolipoprotein, apoE is known to play an important role in lipid transport, and apoE genetic polymorphism has been shown to be associated with susceptibility to Alzheimer's disease. Both apoD and apoE are expressed in the central nervous system (CNS) and both proteins accumulate at sites of peripheral nerve injury due to increased local synthesis. The two proteins may have overlapping or complementary functions within nervous tissue. In order to define the role of apoD within the CNS, we have studied the regional distribution of apoD and apoE mRNA and protein within the normal rat brain and the changes in apoD and apoE expression in the hippocampus of rats after entorhinal cortex lesion (EC lesion). Within the brains of normal rats, apoD expression in the hippocampus was as high as 180-fold that of the liver. ApoD mRNA levels in other areas of the rat brain ranged from 40 to 120 times the hepatic levels. The distribution of apoE gene expression within the brain was similar to that of apoD, but was much lower than hepatic apoE expression. When rats were subjected to EC lesion, the apoD message increased by 54% at 4 days post lesion (DPL) in the ipsilateral region of hippocampus while apoE mRNA levels (ipsilateral and contralateral) decreased by 43%. At 6 to 8 DPL apoD mRNA in the ipsilateral hippocampus remained elevated (42% above controls) whereas the apoE mRNA levels increased to about 15% above those of controls. At 14 and 31 DPL, both apoD and apoE expression was similar to controls. The increase in immunoreactive apoD in hippocampal extracts was more dramatic. At 1 DPL, immunoreactive apoD levels were already 16-fold higher than those in extracts of non-lesioned animals and, at 31 DPL, levels were still 8-fold higher than those of control animals. Finally, we have demonstrated that the levels of apoD in the brains of apoE-deficient mice are 50-fold those of wildtype control mice. ApoD clearly has an important function within the CNS in both normal and pathological situations. PMID- 10381541 TI - Po gene expression is modulated by androgens in the sciatic nerve of adult male rats. AB - The present results show that androgens are able to modulate the Po gene expression in different models. In particular, we have shown that: (1) the messenger for the androgen receptor (AR) is present in the rat sciatic nerve but not in cultured Schwann cells; (2) castration induces a decrease of Po mRNA levels in the sciatic nerve of male rats, which is counteract by the subsequent treatment with dihydrotestosterone (DHT), the 5alpha-reduced metabolite of testosterone; (3) castration is also able to significantly decrease in the sciatic nerve the activity of the enzyme 5alpha-reductase (which converts testosterone into DHT); and (4) DHT is able to stimulate Po gene expression in cultured Schwann cells. These observations seem to indicate that androgens may exert their effect on Po gene expression via indirect mechanisms; modulation of neuronal influences reaching the Schwann cells through the binding of the androgen to the AR present in neurons may be postulated. However, alternative mechanisms may also be taken in consideration. The data presented suggest indeed that androgens might act on Schwann cells via the progesterone receptor (PR) rather than the AR. It has been observed that: (1) the messenger for PR is present in Schwann cells; (2) DHT may activate the transcriptional activity of a PR-responsive gene by binding to the PR; and (3) putative steroid responsive elements have been described in this paper to be present in the Po promoter region. PMID- 10381542 TI - Lithium and valproate differentially regulate brain regional expression of phosphorylated CREB and c-Fos. AB - Previous studies in our laboratory have shown that the mood stabilizers, lithium and valproate (VPA), regulate the transcription factors, cyclic AMP responsive element binding protein (CREB), c-Fos and c-Jun, differentially in cultured human neuroblastoma SH-SY5Y cells. Here, we confirm these findings in rat brain and further study the brain-regional effects of these drugs using immunohistochemistry. We found that although chronic treatment with LiCl or VPA did not change the expression of c-Fos and c-Jun, acute treatment with either drugs increased c-Fos expression but not c-Jun expression in CA1 and CA3 regions of hippocampus. Chronic treatment with LiCl, but not VPA, decreased CREB phosphorylation in rat cerebral cortex and hippocampus. These results suggest that lithium and VPA may act on different pathways to bring about their long-term prophylactic effects on bipolar disorder (BD). The regulation of CREB phosphorylation may be relevant to lithium effect. VPA, which is also effective in BD, may be linked to other pathways. PMID- 10381543 TI - Localization of promoter elements in the human mu-opioid receptor gene and regulation by DNA methylation. AB - The regulation of mu-opioid receptor gene expression was investigated using several molecular techniques. Genomic clones containing portions of the human mu opioid receptor gene were sequenced. 5'-RACE analysis of human brain cDNA confirmed the presence of mRNAs up to -313 from the start codon. As was found for the mouse and rat genes, transcription apparently initiates in the absence of a discernable TATA box. To characterize promoter function, portions of the 5' flanking region were linked to a reporter gene in transient transfection experiments. Two approximately 50 bp adjacent segments had potent, orientation specific promoter activity. More down-stream segments also had promoter activity. None of the 5'-flanking region constructs showed tissue specificity. The potential role of DNA methylation in preventing ectopic expression was investigated by surveying the methylation state of a CpG rich region straddling the start codon. A neural derived cell line (SH-SY5Y) that expresses the mu opioid receptor lacked virtually any CpG methylation. In contrast, two neural derived cell lines that do not express the mu-opioid receptor were nearly totally methylated while non-neural cell lines had intermediate levels of CpG methylation. Additional transient transfection experiments revealed that CpG methylation of the 5'-flanking region suppressed reporter gene expression. These results indicate that CpG methylation plays an important role in regulating mu opioid receptor expression in neural cells; however, no association was found with regulation of expression in non-neural cells. PMID- 10381544 TI - Identification of a novel lithium regulated gene in rat brain. AB - Differential display PCR was used to identify genes regulated by mood stabilizer lithium in rat cerebral cortex. A differentially displayed lithium regulated gene fragment was isolated in rat cerebral cortex after chronic treatment with lithium (1.69 g/kg, p.o. ) for three weeks. A 1216-nucleotide cDNA for a novel lithium regulated gene (NLRG) was isolated from a rat brain cDNA library with RACE (rapid amplification of 5' cDNA end) PCR using a prime from the differentially displayed NLRG gene fragment. The deduced protein sequence was 321 amino acids long, and shows a significant homology with yeast nitrogen permease regulator 2 (NPR2). NLRG expression induced by lithium was confirmed by Northern and slot blot analysis in rat cerebral cortex and neuroblastomaxglioma NG108-15 cells, respectively. In situ hybridization revealed that chronic treatment with lithium increased NLRG gene expression in frontal cortex and hippocampus, but not in striatum, hypothalamus and thalamus regions of rat brain. These results suggest a novel target for lithium which may be relevant to its mechanism of action. PMID- 10381545 TI - Locomotor-activity-induced changes in striatal levels of preprotachykinin and preproenkephalin mRNA. Regulation by the dopaminergic and glutamatergic systems. AB - The mechanisms by which dopaminergic and glutamatergic inputs interact to regulate striatal neuropeptide expression during physiological motor activity are poorly understood. In this work, striatal expression of preprotachykinin (PPT) and preproenkephalin (PPE) mRNA was studied by in situ hybridization in rats killed 2 h after treadmill running (36 m/min for 20 min). Treadmill running induced a significant increase in the levels of both PPT (60% increase) and PPE (90% increase) mRNA in the striatum of normal rats. The increase in the level of PPT mRNA was blocked in rats previously subjected to nigrostriatal deafferentation (i.e., 6-hydroxydopamine lesion) or pretreated with D1-receptor antagonist SCH-23390 (0.1 mg/kg), the D2-receptor antagonist eticlopride (0.5 mg/kg), or the N-methyl-D-aspartate (NMDA) glutamate receptor antagonist MK-801 (0.1 mg/kg). The running-induced increase in the level of PPE mRNA was blocked in rats pretreated with SCH-23390 or MK-801. Rats subjected to nigrostriatal deafferentation or pretreated with eticlopride showed an increase in PPE mRNA levels (around 150% and 40% increase, respectively), that was enhanced by running (around 230% and 160% increase, respectively). These results suggest that locomotor activity increases, in a NMDA receptor dependent fashion, the excitatory influence of the corticostriatal glutamatergic system on the two populations of striatal projection neurons, as reflected by increases in the levels of PPT and PPE mRNA. The results obtained after dopamine depletion or injection of dopamine receptor antagonists suggest that a concomitant increase in dopamine release may enhance PPT mRNA level in striatonigral neurons via D1 receptors, and reduce PPE mRNA level in striatopallidal neurons via D2 receptors. Additionally, levels of dopamine and glutamate may be regulated by other complex indirect mechanisms. PMID- 10381547 TI - Basal and IL-1beta-stimulated cytokine and neuropeptide mRNA expression in brain regions of young and old Long-Evans rats. AB - Young and old Long-Evans rats respond with fevers of equal magnitude and duration to the brain administration of interleukin-1beta (IL-1beta). Here, we characterized brain regional mRNA expression of cytokine and neuropeptide components in response to the brain administration of IL-1beta. We used specific and highly sensitive RNase protection assays to determine mRNA changes for IL 1beta, IL-1 receptor type I (IL-1RI), IL-1R accessory proteins I and II (IL-1R AcP I and II), IL-1 receptor antagonist (IL-1Ra), transforming growth factor beta1 (TGF-beta1), glycoprotein 130 (gp 130), leptin receptor (OB-R), neuropeptide Y (NPY) and pro-opiomelanocortin (POMC) in the cerebellum, parieto frontal cortex, hippocampus, hypothalamus, and midbrain of male young (3-5 months) and old (24-26 months) Long-Evans rats. In both young and old rats, IL 1beta induced a significant up-regulation of cerebellar IL-1Ra, IL-1RI, and TGF beta1 mRNAs; hippocampal TGF-beta1 mRNA; hypothalamic IL-1beta, IL-1Ra, TGF beta1, and gp 130 mRNAs; and midbrain IL-1beta and TGF-beta1 mRNAs. There were no age-related differences in any cytokine mRNA levels under basal or IL-1beta stimulated conditions. Levels of hypothalamic POMC mRNA were different between age groups under basal and stimulated conditions. IL-1R AcP I and leptin receptor did not change in any brain region from either young or old rats, suggesting specificity of transcriptional changes. The data show that old Long-Evans rats are not defective in their capacity to develop an appropriate cytokine response to the brain administration of IL-1beta. The implications of these findings for neuroimmunological-neuroinflammatory and neurotoxic/neurodegenerative processes are discussed. PMID- 10381546 TI - Calcium influx and membrane depolarization induce phosphorylation of neurofilament (NF-M) KSP repeats in PC12 cells. AB - Signals activating the kinases that phosphorylate neurofilament proteins in the axon remain unknown. In a previous study, we have demonstrated that a constitutively active form of MEK1 activates Erk1 and Erk2 kinases, which phosphorylate co-transfected NF-M in NIH 3T3 cells. In this study, we report the activation of endogenous Erk1 and Erk2 by membrane depolarization and calcium influx through L-type calcium channels, which resulted in phosphorylation of the NF-M tail domain in PC12 cells. This phosphorylation was inhibited in the presence of nifedipine, an L-type calcium channel inhibitor, and PD98059, a specific MEK1 inhibitor. Our data suggest a mechanism linking calcium influx through voltage-gated calcium channels with the MAP kinase pathway and NF-M tail domain phosphorylation in cell body and neurite. These findings may provide significant new insights into mechanisms involved in some neurological diseases. PMID- 10381548 TI - Characterization of multiple forms of the human glycine transporter type-2. AB - The human glycine transporter type 2 (hGlyT2) was cloned from a spinal cord cDNA library using PCR-based methodologies. The isolated sequence exhibits 89% homology with the previously isolated rat GlyT2 cDNA (Liu et al., J. Biol. Chem. 268 (1993) 22802-22808) at the nucleotide level, and 93% amino acid sequence identity. The greatest divergence between the human and rat sequences is found at the amino-terminus, where only 74% amino acid identity exists in residues 1-190. Expression of the intact hGlyT2 transporter sequence in COS-7 cells resulted in a 10-fold increase in high-affinity uptake relative to control cells transfected with vector alone. An artificially truncated form of the transporter, missing the NH2-terminal 153 amino acids, was also capable of mediating glycine uptake. However, an identified variant lacking the first 234 amino acids was non functional. An hGlyT2 transporter containing a 14-residue deletion in the intracellular loop between transmembrane domains 6 and 7 was also identified and expressed, but failed to mediate glycine uptake. Like rat GlyT2, the high affinity uptake mediated by hGlyT2 was found to be insensitive to the GlyT1 inhibitor sarcosine. PMID- 10381549 TI - Distinct localization of SAPK isoforms in neurons of adult mouse brain implies multiple signaling modes of SAPK pathway. AB - Various cellular and environmental stresses lead to the activation of stress activated protein kinase (SAPK), which is also referred to as c-Jun N-terminal kinase (JNK). In mammals, multiple SAPK isoforms, encoded by three independent genes, were identified. To gain insight into the roles of SAPK pathway in adult mouse brain, detailed expression patterns of three SAPK isoforms in brain were examined by using immunohistochemical and cell biological analyses. SAPKbeta was heavily expressed in almost all regions of brain as previously reported. Interestingly, SAPKgamma was also widely expressed at high levels. SAPKgamma expression was generally overlapped with SAPKbeta although there were some exceptions such as in hippocampus, where SAPKgamma was restricted to CA3 and CA4 regions while SAPKbeta was evenly expressed. SAPKalpha was widely expressed, but at low levels. It is particularly intriguing to note the differential subcellular localization of SAPK isoforms in neurons. In brain of normally reared mice, SAPKbeta was identified in nucleus as well as in cytoplasm of neurons, while SAPKgamma was detected mainly in cytoplasm and dendrites. Biochemical and immunological analyses revealed extraordinarily high basal activities of all SAPK isoforms in brain compared to peripheral organs, indicating that SAPK pathway may play a role in normal brain physiology. In addition, differential regional and subcellular localizations of SAPK isoforms allow us to speculate multiple signaling modes for SAPK activation in brain. PMID- 10381550 TI - Synergistic interaction between serotonin-2 receptor and dopamine D1 receptor stimulation on striatal preprotachykinin mRNA expression in the 6-hydroxydopamine lesioned rat. AB - The regulation of striatal preprotachykinin (PPT) mRNA expression can be mediated through both dopamine (DA) D1 and serotonin (5-HT) 5-HT2A/2C receptors. In the present study, we used in situ hybridization to examine possible synergistic interactions between 5-HT2A/2C and D1 receptor-mediated regulation of striatal PPT mRNA levels in the rat depleted of DA with 6-hydroxydopamine. Acute administration of the 5-HT2A/2C receptor agonist DOI (2 mg/kg) significantly increased (+75%) PPT mRNA levels in the dorsal striatum. Acute administration of the D1 receptor agonist SKF-38393 (2 mg/kg) did not significantly alter PPT mRNA levels in the dorsal striatum. However, the co-administration of SKF-38393 and DOI produced a significant increase (+300%) in striatal PPT mRNA expression restricted to the periventricular region of the dorsal-medial striatum. This synergistic interaction was not observed in the remaining aspect of the dorsal striatum where DOI alone increased PPT mRNA expression. These data show that 5 HT2A/2C and D1 receptors can act in a synergistic manner to regulate striatal PPT mRNA in a subregion of the DA-depleted striatum. PMID- 10381551 TI - Selective expression of serotonin receptor transcripts in the mammalian cochlea and its subdivisions. AB - Expression of serotonin receptor (5-HTR) mRNA has been determined in the mammalian cochlea and its subdivisions by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. Specific primers targeting individual 5-HTRs 1-7 directed amplification of 5-HTR subtypes 1A, 1B, 2B, 2C, 3, 5B, and 6 from mouse cochlea cDNA. No evidence of expression was obtained for 5-HTRs 1D, 2A, 4 (L and S), 5A, and 7. The distribution of receptor mRNA within the cochlea was determined with application of RT-PCR to morphologically defined microdissected subfractions of the rat cochlea. Messages for 5-HTR subtypes 1A, 1B, 2B, and 6 were present in the organ of Corti, lateral wall, and spiral ganglion subfractions. Messages for 5-HTR subtypes 2C, 3 and 5B were found in the spiral ganglion, but not in the organ of Corti or lateral wall fractions. The existence of transcripts for 5-HTRs 1A, 1B, 2B and 6 in the organ of Corti is consistent with a role for these receptors in serotonin-mediated modulation of the mechanosensory signal. PMID- 10381552 TI - Expression of the thymosin beta10 gene in normal and kainic acid-treated rat forebrain. AB - Thymosin beta10 (Tbeta10) is a small actin-sequestering peptide widely distributed in mammalian tissues including nervous system. Here, we analyze the expression of Tbeta10 gene in normal and kainic acid (KA)-treated rat forebrain by in situ hybridization. Our results showed a defined regional pattern of the mRNA encoding for Tbeta10 in both normal and KA-treated rat forebrain. The presence of this transcript in different regions of the rat forebrain, including hippocampus, neocortex and several brain nuclei, provides evidence for the participation of Tbeta10 in the control of the actin dynamics that takes place in neurons. Furthermore, the analysis of the forebrain in KA-treated rats revealed an activation of the Tbeta10 gene linked to gliosis. PMID- 10381553 TI - Developmental expression of the CaM kinase II isoforms: ubiquitous gamma- and delta-CaM kinase II are the early isoforms and most abundant in the developing nervous system. AB - CaM kinase II constitutes a family of multifunctional protein kinases that play a major role in Ca2+-mediated signal transduction. As a first step in understanding their possible function in mouse development we characterized the expression patterns of all CaM kinase II isoforms (alpha, beta, gamma and delta) starting in prenatal development. Remarkably, only the ubiquitous gamma- and delta-CaM kinase II are expressed during early development. Their distribution suggests a special role in the developing nervous system and in mature excitable tissues. Additionally, we describe the murine betaM-CaM kinase II, a variant of the 'brain specific' beta-CaM kinase II, which is highly expressed in skeletal muscle. PMID- 10381554 TI - Mu-opioid receptor mRNA expression in proopiomelanocortin neurons of the rat arcuate nucleus. AB - It has been previously demonstrated that the activity of proopiomelanocortin (POMC)-containing neurons in the rat arcuate nucleus is regulated by opiates, but the expression of opioid receptors in POMC neurons has never been reported. In the present study, we have applied a double-labeling in situ hybridization technique to investigate the occurrence of mu-opioid receptor mRNA on POMC neurons. We have found that 20+/-3% of arcuate POMC neurons express mu-opioid receptor mRNA and that the proportion of POMC neurons expressing mu-opioid receptor is higher in the caudal than in the rostral portion of the arcuate nucleus. Our data suggest that POMC neurons might be both auto-regulated by beta endorphin, and regulated by enkephalins. PMID- 10381555 TI - Caspase-3-dependent neuronal death in the hippocampus following kainic acid treatment. AB - In this study, we examined the levels of activated caspase-3 in the kainic acid (KA) model of hippocampal degeneration in both sensitive (FVB/N) and resistant (129/SvEMS) strains of mice. At 30 h, 2 and 4 days following KA administration, animals were sacrificed and brains examined for pyknosis, TUNEL labeling, and activated caspase-3 immunoreactivity. Catalytically active caspase-3 was first detected 30 h following KA treatment in the sensitive, FVB/N strain. This was 18 h before the appearance of pyknosis or TUNEL labeling. The expression of activated caspase-3 continues up to 4 days post-injection. No activated caspase-3 immunoreactivity was detected in the resistant, 129/SvEMS strain, neither was there evidence of pyknosis or TUNEL staining. This suggests that activation of caspase-3 is a necessary component of KA-induced cell death. PMID- 10381556 TI - Induction of cerebellar hsp72 in rats learning a two-way active avoidance task. AB - The involvement of brain heat shock proteins in learning was examined by Western analyses in rats trained for an active avoidance task, and in passive and active controls. Expression of the constitutive hsp73 was intense in brain, liver, and kidney of all rats. Conversely, expression of the inducible hsp72 occurred in the cerebellum of most trained rats, but not in passive or active controls. Significant correlations were present between avoidances and cerebellar scores determined 8 h after training. Induction of hsp72 may therefore be attributed to learning in the cerebellum, while in other brain regions, liver and kidney stress related stimuli may play a prevalent role. PMID- 10381557 TI - Bcl-2 expression regulates cell sensitivity to S100beta-mediated apoptosis. AB - The S100beta protein is overexpressed in the brain of patients with Alzheimer's disease and Down's syndrome and is able to induce apoptosis in neurons at high concentrations. The intracellular events that regulate the apoptotic effect are largely unknown. This study investigates the roles of the bcl-2 proto-oncogene, one of the best-defined apoptotic genes, on cell death induced by S100beta. Human neuronal precursor NT2/D1 cells showed a high degree of cell death by apoptosis after exposure to 2 microM S100beta in serum-free medium. Death was preceded by a down-regulation of the Bcl-2 protein. Gene transfer with a full-length bcl-2 cDNA under the control of a constitutive promoter in NT2 cells elevated Bcl-2 protein levels and repressed S100beta-mediated cell death. When exposed to retinoic acid, the NT2/D1 cells differentiated into a neuronal phenotype. The differentiated cells up-regulated their levels of Bcl-2 and became resistant to S100beta-induced cell death. Downregulation of Bcl-2 by an antisense oligonucleotide in the differentiated cells, however, increased their susceptibility to S100beta-related cytotoxicity. Therefore, apoptosis induced through S100beta signaling is subject to regulation by Bcl-2. A combined alteration such as up-regulation of S100beta together with down-regulation of Bcl-2 may be important in the pathogenesis of Alzheimer's disease and Down's syndrome. PMID- 10381558 TI - Blood sugar formation due to abnormally elevated gluconeogenesis: aberrant regulation in a parasitized insect, Manduca sexta Linnaeus. AB - Alterations of carbohydrate metabolism associated with parasitism were examined in an insect, Manduca sexta L. In insect larvae maintained on a low carbohydrate diet gluconeogenesis from [3-13C]alanine was established from the fractional 13C enrichment in trehalose, a disaccharide of glucose and the blood sugar of insects and other invertebrates. After transamination of the isotopically substituted substrate to [3-13C]pyruvate, the latter was carboxylated to oxaloacetate ultimately leading to de novo glucose synthesis and trehalose formation. Trehalose was selectively enriched with 13C at C1 and C6 followed by C2 and C5. 13C enrichment of blood sugar in insects parasitized by Cotesia congregata (Say) was significantly greater than was observed in normal animals. The relative contributions of pyruvate carboxylation and decarboxylation to trehalose labeling were determined from the 13C distribution in glutamine, synthesized as a byproduct of the tricarboxylic acid cycle. The relative contribution of carboxylation was significantly greater in parasitized larvae than in normal insects providing additional evidence of elevated gluconeogenesis due to parasitism. Despite the increased gluconeogenesis in parasitized insects the level of blood sugar was the same in all animals. Because de novo glucose synthesis does not normally maintain blood sugar level in insects maintained under these dietary conditions the findings suggest an aberrant regulation over gluconeogenesis. The 13C labeling in trehalose was nearly symmetric in all insects but the mean C1/C6 13C ratio was higher in parasitized animals suggesting a lower activity of the pentose phosphate pathway that brings about a redistribution of 13C in trehalose following de novo glucose synthesis. Additional studies with insects maintained on a high carbohydrate diet and administered [1,2-13C2]glucose confirmed a decreased level of pentose cycling during parasitism consistent with a lower level of lipogenesis. It is suggested, however, that the pentose pathway may facilitate the synthesis of trehalose from dietary carbohydrate by directing hexose phosphate cycled through the pathway to the production of energy. PMID- 10381559 TI - In vitro and in vivo studies investigating possible antioxidant actions of nicotine: relevance to Parkinson's and Alzheimer's diseases. AB - An inverse relationship appears to exist between cigarette smoking and the risk of Parkinson's and Alzheimer's diseases. Since both diseases are characterized by enhanced oxidative stress, we investigated the antioxidant potential of nicotine, a primary component of cigarette smoke. Initial chromatographic studies suggest that nicotine can affect the formation of the neurotoxin 6-hydroxydopamine resulting from the addition of dopamine to Fenton's reagent (i.e., Fe2+ and H2O2). Thus, under certain circumstances, nicotine can strongly affect the course of the Fenton reaction. In in vivo studies, adult male rats being treated with nicotine showed greater memory retention than controls in a water maze task. However, neurochemical analysis of neocortex, hippocampus, and neostriatum from these same animals revealed that nicotine treatment had no effect on the formation of reactive oxygen species or on lipid peroxidation for any brain region studied. In an in vitro study, addition of various concentrations of nicotine to rat neocortical homogenates had no effect on lipid peroxidation compared to saline controls. The results of these studies suggest that the beneficial/protective effects of nicotine in both Parkinson's disease and Alzheimer's disease may be, at least partly, due to antioxidant mechanisms. PMID- 10381560 TI - Kinetics of hemin distribution in plasma reveals its role in lipoprotein oxidation. AB - Hemin is a powerful in vitro inducer of low-density lipoprotein (LDL) oxidation, implicated in development of atherosclerosis. To support the proposed role of hemin in atherogenesis, the question of whether hemin has any chance of getting together with LDL in vivo, must be addressed. A stopped-flow technique was employed in order to investigate the fast kinetics of hemin binding to LDL and to other plasma hemin-binding proteins: high-density lipoprotein (HDL), albumin and hemopexin. Based on the measured rate constants of hemin association with and dissociation from each of these proteins, time-dependent hemin distribution in plasma was analyzed. The analysis shows that as much as 80% of total hemin binds initially to LDL and HDL, the plasma components which are most susceptible to oxidation. Only then hemin partially transfers to the antioxidants albumin and hemopexin. The half time of the hemin-LDL complex in plasma, initially comprising 27% of total hemin, was more than 20 s. Not only transient, but also oxidatively active steady-state hemin-lipoprotein complexes in plasma were both predicted from the kinetic analysis and found in experiment. Our data suggest that the hemin-LDL complex may exist in vivo and that its oxidative potential should be considered pro-atherogenic. PMID- 10381561 TI - Receptor/ligand interactions between Cryptosporidium parvum and the surface of the host cell. AB - The ability of membrane antigens on sporozoites of the intestinal pathogen, Cryptosporidium parvum, to bind host cell surface antigens was investigated. A novel membrane-associated protein of approximately 47 kDa, designated CP47, was found to possess significant binding affinity for the surface of both human and animal ileal cells. This protein was purified by a combination of anion-exchange chromatography on FPLC and immunoaffinity chromatography. Purified CP47 demonstrated competitive binding with parasite-associated membrane antigens to membranes of HCT-8 and ileal cells in a dose-dependent manner. Furthermore, the binding activity of CP47 was found to be Mn2+-sensitive, and was completely inhibited in the presence of 10 mM MnCl2. These results were consistent with earlier findings demonstrating the inhibitory effect of Mn2+ ions on Cryptosporidium infection both in vitro and in vivo (Nesterenko et al., Biol. Trace Elem. Res. 56 (1997) 243-253). Immunoelectron microscopy using gold conjugated antibodies revealed CP47 to be localized at the apical end of the sporozoites. A single protein with an electrophoretic mobility of 57 kDa was purified from host cell membranes using CP47-Affigel. Similarly, affinity purification of this protein was abrogated in the presence of Mn2+. These data suggest that a novel parasite protein, CP47, may play an important role in sporozoite/host cell attachment. PMID- 10381562 TI - Enhanced channelling of sulphate through a rapidly exchangeable sulphate pool in response to stimulated glycosaminoglycan synthesis in pancreatic epithelial cells. AB - The ability of cells to decorate glycosaminoglycans (GAGs) with sulphate in highly specific patterns is important to extracellular matrix biogenesis and placing appropriate glycosulphated ligands on the cell surface. We have examined sulphate metabolism in two pancreatic duct epithelial cell lines - PANC-1 and CFPAC-1 (derived from a cystic fibrosis patient) with a view to understanding how pancreatic cells utilise intracellular sulphate. [35S]Sulphate uptake was rapid and reached near steady state levels within 10 min. However, the intracellular specific activity of [35S]sulphate for PANC-1 and CFPAC-1 reached only 35 and 10%, respectively, of the medium specific activity at 10 min. Therefore, sulphate appears to reside within two compartments; a rapidly exchangeable sulphate pool (RESP) and a slowly exchangeable sulphate pool (SESP). Reducing chloride in the medium, increased the specific activity of [35S]sulphate within cells and increased the size of the inorganic sulphate pool, suggesting that the RESP was enlarged. Sulphate pools were not different in size between the two cell lines in physiological NaCl. Increasing the size of the sulphate pool had no effect on [35S]sulphate:[3H]glucosamine ratios incorporated into glycosaminoglycans (GAGs); however, stimulating the synthesis of GAGs with 4-methylumbelliferyl-beta-d xyloside, stably elevated [35S]:[3H] ratios. This was due to higher [35S]sulphate incorporation. [35S]Cysteine contributed less than 0.1% of the cells' sulphate requirements. We conclude that in the face of elevated demand for sulphate, pancreatic cells appear to channel a greater proportion through the RESP. PMID- 10381563 TI - Inactivation and loss of antigenicity of esterase by sugars and a steroid. AB - Glycation, the non-enzymic reaction of sugars with proteins, has an important role in the complications of diabetes. It has been studied mostly in structural proteins but more recently has been shown to inactivate enzymes. Previous evidence from our laboratory indicated that glycation-induced inactivation and loss of antigenicity of catalase and superoxide dismutase are simultaneous. Esterase, which decreases activity in the lens in senile cataract and diabetes, was measured by a spectrophotometric assay using p-nitrophenyl acetate as the substrate. Here we investigated the inactivation of carboxylesterase (EC 3.1.1.1) by sugars of different glycating power and prednisolone-21-hemisuccinate while simultaneously monitoring the loss of antigenicity. Antigenicity was assessed by immunoprecipitation and by dot-blotting the glycated and non-glycated fractions of enzymes separated by affinity chromatography. Ribose and fructose inactivated more rapidly than glucose and glucose 6-phosphate. The esterase was progressively inactivated by prednisolone-21-hemisuccinate at a lower concentration. Activity and antigenicity were lost simultaneously. The glycated enzyme had entirely lost its antigenicity. These results further support the idea that inactivation of enzyme and loss of antigenicity are simultaneous. PMID- 10381564 TI - The effect of prolonged iron loading on the chemical form of iron oxide deposits in rat liver and spleen. AB - Female Porton rats were loaded with iron either by supplementing the diet with 2.5% carbonyl iron for up to 22 months (18 rats) or by regularly injecting rat blood cells intraperitoneally for up to 10 months (eight rats). 57Fe Mossbauer spectroscopy of freeze-dried samples of liver and spleen was used to analyse the chemical forms of iron deposited in these tissues over the period of iron loading. A sextet signal in the Mossbauer spectra was identified as being due to a form of haemosiderin based on the structure of the mineral goethite. The spectral parameters of the sextet signal in the rat tissues indicate that the goethite-like haemosiderin particles are less crystalline than those found in iron-loaded human tissues. For the dietary-iron-loaded rat livers, the fraction (Fs) of the Mossbauer signal in the form of this sextet was found to increase significantly (from approx 0.04 to 0.09) with the age of the rats (r=0.77, P<0.0005). This indicates that the fraction of liver iron in the form of the goethite-like haemosiderin increases with age of the rat and hence with the duration of iron loading. In addition, Fs for these livers was found to increase significantly with the fraction of iron in non-parenchymal cells as measured by computer-assisted morphometric analysis of histological sections (r=0.71, P<0.005). PMID- 10381565 TI - Cloning and expression of Der f 6, a serine protease allergen from the house dust mite, Dermatophagoides farinae. AB - House dust mite allergen is thought to be a major cause of asthma. Characterization of these allergen molecules is therefore an important step for the development of effective diagnostic and therapeutic agents against mite associated allergic disorders. Here we report molecular cloning and expression of the group 6 (chymotrypsin-like) allergen from the house dust mite, Dermatophagoides farinae. Sequencing analysis indicates that cloned cDNA, designated Der f 6, encodes a 279 amino acid polypeptide which conserves a primary structure characteristic for chymotrypsin-like serine proteases found in mammals. Recombinant Der f 6 expressed in Escherichia coli bound IgE in a pool made of 20 sera, and induced histamine release from patients' peripheral blood cells. PMID- 10381567 TI - The PBX-regulating protein PREP1 is present in different PBX-complexed forms in mouse. AB - Human PREP1, a novel homeodomain protein of the TALE super-family, forms a stable DNA-binding complex with PBX proteins in solution, a ternary complex with PBX and HOXB1 on DNA, and is able to act as a co-activator in the transcription of PBX HOXB1 activated promoters (Berthelsen, J., Zappavigna, V., Ferretti, E., Mavilio, F., Blasi, F. , 1998b. The novel homeoprotein Prep1 modulates Pbx-Hox protein cooperatity. EMBO J. 17, 1434-1445; Berthelsen, J., Zappavigna, V., Mavilio, F., Blasi, F., 1998c. Prep1, a novel functional partner of Pbx proteins. EMBO J. 17, 1423-1433). Here we demonstrate the presence of DNA-binding PREP1-PBX complexes also in murine cells. In vivo, PREP1 is a predominant partner of PBX proteins in various murine tissues. However, the choice of PBX family member associated with PREP1 is largely tissue-type specific. We report the cloning and expression domain of murine Prep1 gene. Murine PREP1 shares 100% identity with human PREP1 in the homeodomain and 95% similarity throughout the whole protein. In the adult mouse, PREP1 is expressed ubiquitously, with peaks in testis and thymus. We further demonstrate the presence of murine Prep1 mRNA and protein, and of different DNA-binding PREP1-PBX complexes, in mouse embryos from at least 9.5 days p.c. Moreover, we show that PREP1 is present in all embryonic tissues from at least 7.5-17.5 days p.c with a predominantly nuclear staining. PREP1 is able to super-activate the PBX-HOXB-1 autoregulated Hoxb-1 promoter, and we show that all three proteins, PREP1, PBX and HOXB-1, are present together in the mouse rhombomere 4 domain in vivo, compatible with a role of PREP1 as a regulator of PBX and HOXB-1 proteins activity during development. PMID- 10381566 TI - N-myc-dependent repression of ndr1, a gene identified by direct subtraction of whole mouse embryo cDNAs between wild type and N-myc mutant. AB - To identify genes regulated by N-myc, subtraction of whole embryo cDNA was carried out between wild type and N-myc-deficient mutant mice. Six cDNA clones were isolated as representing genes expressed higher in the mutant embryos and two as those expressed lower. One of them, Ndr1, coding for 43 kDa cytoplasmic protein was studied in detail. The Ndr1 gene was augmented 20-fold in the mutant embryos at 10.5 days post coitus which is indicative of repression by N-myc. An inverse relationship actually existed between the expression of N-myc and Ndr1 in various developing tissues of the wild type embryos. In the early stage of differentiation of these tissues when N-myc expression was high Ndr1 expression was low or undetectable, and later when N-myc activity diminished Ndr1 expression was augmented concomitantly with the occurrence of terminal differentiation. To establish the direct link between N-myc activity and the Ndr1 regulation, the Ndr1 gene was cloned and analyzed. The Ndr1 promoter activity was down-regulated by N-myc, and more strongly by the combination of N-myc and Max in the cotransfection assay. This repressive effect was mediated by the promoter region within 52 base pairs from the transcription start site but direct binding of N myc:Max to the promoter sequence was not demonstrated, which is analogous to the cases recently reported for transcriptional repression by c-myc. c-myc also repressed Ndr1 promoter activity similarly to N-myc. The effect of N-myc:Max was sensitive to Trichostatin A, indicating involvement of histone deacetylase activity in repression of the Ndr1 promoter. The strategy we adopted in identifying target genes of a transcription factor should prove widely applicable when mutant animals are available. PMID- 10381568 TI - Divergent properties of mouse netrins. AB - The netrins are a small but highly conserved family of axonal guidance signals found throughout the animal kingdom. This sequence conservation was used to isolated cDNAs for two mouse netrins. Analysis of their expression patterns and functional properties showed that mouse netrin-1 is in most respects similar to its orthologs in other vertebrates while the properties of netrin-3 differ markedly from those of other members of this protein family. In contrast to netrin-1 which is widely expressed both in the developing nervous system and in mesodermal tissues, netrin-3 transcripts are largely restricted to dorsal root ganglia and the developing limb buds. Netrin-3 binds with a significantly lower affinity to the netrin receptor DCC (deleted in colorectal cancer) and is also ineffective in eliciting the outgrowth of commissural axons in vitro. These results demonstrate that, although the netrins are highly conserved signals that guide axons to or away from the midline of the developing nervous system, at the same time they show a surprising degree of divergence in vertebrates. PMID- 10381569 TI - An early developmental role for eph-ephrin interaction during vertebrate gastrulation. AB - Eph receptor tyrosine kinases (RTK) and their ephrin ligands are involved in the transmission of signals which regulate cytoskeletal organisation and cell migration, and are expressed in spatially restricted patterns at discrete phases during embryogenesis. Loss of function mutants of Eph RTK or ephrin genes result in defects in neuronal pathfinding or cell migration. In this report we show that soluble forms of human EphA3 and ephrin-A5, acting as dominant negative inhibitors, interfere with early events in zebrafish embryogenesis. Exogenous expression of both proteins results in dose-dependent defects in somite development and organisation of the midbrain-hindbrain boundary and hindbrain. The nature of the defects as well as the distribution and timing of expression of endogenous ligands/receptors for both proteins suggest that Eph-ephrin interaction is required for the organisation of embryonic structures by coordinating the cellular movements of convergence during gastrulation. PMID- 10381571 TI - Toll homologue expression in the beetle tribolium suggests a different mode of dorsoventral patterning than in drosophila embryos. AB - The gene Toll (Tl) encodes a maternally supplied interleukin 1 receptor-related transmembrane protein, a key component required to establish dorsoventral polarity in the Drosophila embryo. We have isolated Tl homologs of a primitive dipteran, Clogmia albipunctata, and of the beetle Tribolium castaneum. Tribolium Tl protein (Tl) lacks sequences in the C-terminal portion of the cytoplasmic domains that are conserved in the dipteran homologs. The Tl homolog of Tribolium mediates the ventralizing activity when expressed as a gain-of-function variant in transgenic Drosophila, indicating that the sequences conserved in the Diptera are not essential for Tl signaling. In contrast to Drosophila where Tl gene expression occurs maternally and supplies uniformly distributed Tl in the egg membrane, Tl transcripts form a ventral-to-dorsal gradient in the Tribolium blastoderm stage embryo. This localized expression pattern of Tl transcripts, as compared with the strong maternal and ubiquitous expression in Drosophila and Clogmia embryos, suggests that dorsoventral patterning in long-germ band and short-germ band insects involves the same components but different modes of their action. PMID- 10381570 TI - A double interaction screen identifies positive and negative ftz gene regulators and ftz-interacting proteins. AB - Regulatory genes directing embryonic development are expressed in complex patterns. The Drosophila homeobox gene fushi tarazu (ftz) is expressed in a striped pattern that is controlled by several discrete and large cis- regulatory elements. One key cis-element is the ftz proximal enhancer which is required for stripe establishment and which mediates autoregulation by direct binding of Ftz protein. To identify the trans-acting factors that regulate ftz expression and autoregulation, we developed a modified yeast two hybrid screen, the Double Interaction Screen (DIS). The DIS was designed to isolate both DNA binding transcriptional regulators that interact with the proximal enhancer and proteins that interact with Ftz itself when it is bound to the enhancer. The screen identified two candidate Ftz protein cofactors as well as activators and repressors of ftz transcription that bind directly to the enhancer. One of these (Tramtrack (Ttk)) was previously shown to bind to at least five sites in the proximal enhancer; genetic studies suggested that Ttk acts as a repressor of ftz in the embryo. Here we show that, in yeast cells, Ttk protein strongly activates transcription, suggesting that yeast may be missing a necessary co-repressor which is present in Drosophila embryos. Further, we have characterized the activity of a second candidate ftz repressor isolated in the screen - the product of the pair-rule gene sloppy paired - a member of the forkhead family. We show that Slp1 is a DNA binding protein. We have identified a high affinity binding site for Slp1 in the ftz proximal enhancer. Slp1 represses transcription via this binding site in yeast cells, consistent with its role as a direct repressor of ftz stripes in interstripe regions during late stages of embryogenesis. The DIS should be a generally useful method to identify DNA binding transcriptional regulators and protein partners of previously characterized DNA binding proteins. PMID- 10381572 TI - The expression and function of cadherin-mediated cell-to-cell adhesion in human embryonal carcinoma cells. AB - Human embryonal carcinoma (EC) cells typically require high cell densities to maintain their characteristic phenotype; they are generally subject to differentiation when cultured at low cell densities, marked by changes in morphology and expression of the surface antigen, SSEA-1. To test whether cadherin mediated cell-to-cell adhesion may be responsible for maintaining an EC phenotype we ascertained that human EC cells generally express E- and P cadherins, and are subject to cadherin mediated, Ca2+ dependent aggregation. However, in the NTERA2 human EC cell line, inhibition of cadherin mediated adhesion by culture in low levels of Ca2+ did not result in the changes typically seen under low cell density conditions. Low Ca2+ levels also did not affect the pattern of differentiation in these cells following induction with retinoic acid. Therefore, cadherin-mediated cell adhesion does not appear to play a role in maintaining an EC phenotype. On the other hand, culture at both low cell density and in the absence of Ca2+ did result in changes in the patterns of cadherin expression suggesting a feedback regulatory effect of cell-to-cell adhesion. Further, lithium which inhibits the cytoplasmic kinase GSK3beta and hence influences beta-catenin levels did cause differentiation of NTERA2 cells. However, consideration of the phenotype of the resultant cells suggested that this effect may be because of lithium mimicking activation of a Wnt signalling pathway, rather than an effect on signalling consequent upon cadherin mediated cell to cell adhesion. PMID- 10381573 TI - Six class homeobox genes in drosophila belong to three distinct families and are involved in head development. AB - The vertebrate Six genes are homologues of the Drosophila homeobox gene sine oculis (so), which is essential for development of the entire visual system. Here we describe two new Six genes in Drosophila, D-Six3 and D-Six4, which encode proteins with strongest similarity to vertebrate Six3 and Six4, respectively. In addition, we report the partial sequences of 12 Six gene homologues from several lower vertebrates and show that the class of Six proteins can be subdivided into three major families, each including one Drosophila member. Similar to so, both D Six3 and D-Six4 are initially expressed at the blastoderm stage in narrow regions of the prospective head and during later stages in specific groups of head midline neurectodermal cells. D-Six3 may also be essential for development of the clypeolabrum and several head sensory organs. Thus, the major function of the ancestral Six gene probably involved specification of neural structures in the cephalic region. PMID- 10381574 TI - Functional determinants for the tetracycline-dependent transactivator tTA in transgenic mouse embryos. AB - Tetracycline-dependent transgenes promise to be an important tool for investigating the time dependence of gene function during mouse development. The pivotal element of this approach is the recombinant tetracycline-dependent transactivator tTA. Using a modified gene trap approach we successfully generated mouse lines expressing tTA in a wide spread manner during embryogenesis. The transgenic model system which we established allowed us to depict transactivator and target gene expression patterns with high resolution by histochemical means. Our data provide evidence that with decreasing concentrations of tTA protein the state of chromatin acetylation becomes an increasingly important determinant for tTA function. The observation of tTA-dependent position effects on tetO-linked target genes suggests that transcription patterns can be encoded at the level of promoter preactivation. PMID- 10381575 TI - Six9 (Optx2), a new member of the six gene family of transcription factors, is expressed at early stages of vertebrate ocular and pituitary development. AB - The Drosophila gene sine oculis (so) is a nuclear homeoprotein, which is required for eye development. Several homologues of so have been found in vertebrates. We report here a detailed expression analysis in chick and mouse of Six9 (Optx2), a novel gene of the Six/sine oculis family closely related to Six3. Six9 (Optx2) is first expressed at presomitic stages in the head-fold, both in the neural plate and in the underlying axial mesoderm. Thereafter, Six9 (Optx2) is strongly expressed in the presumptive and differentiating neural retina and ventral optic stalk, in the olfactory placodes, in the hypothalamus and in the pituitary gland. This expression pattern largely overlaps with that of Six3, but several differences exist between the expression domain of the two genes. At presomitic stages, the posterior boundary of Six3 expression is at the same axial level both in the prechordal plate and in the overlying neural plate. In contrast, Six9 (Optx2) expression in the prechordal plate extends more caudal to that of the neural plate, occupying a more restricted V-shaped territory. Similarly, during the early events of eye patterning, Six3 is first expressed in the entire optic vesicle and lens placode. Only later does its expression become confined to the prospective and differentiating neural retina. Conversely, Six9 (Optx2) is never observed in the lens placode of either chick and mouse, and from early stages of optic vesicle development, Six9 (Optx2) transcripts are restrained to the prospective ventral neural retina and optic stalks. PMID- 10381576 TI - Differential expression of KDR/VEGFR-2 and CD34 during mesoderm development of the early human embryo. AB - Recent findings on vertebrate embryos have provided compelling evidence for the existence of hemangioblasts, i.e. common precursors for endothelial and hematopoietic cells, characterized by expression of the VEGFR2/Flk1 receptor. We describe here a population of KDR+ CD34- mesoderm cells that emerges in early somitic human embryos, by the beginning of the 4th week of gestation. In the developing blood vessels, KDR-expressing CD34- cells gradually coexpress increasing levels of CD34 antigen. Remarkably, as development proceeds, a KDR+ CD34- contingent persists in the paraaortic splanchnopleura until just prior to the emergence of aorta-associated hematopoietic cell clusters. These observations suggest that KDR+ CD34- mesodermal cells might represent the putative hemangioblastic precursor of human hematopoietic and endothelial lineages. PMID- 10381577 TI - Genomic structure, mapping, activity and expression of fibroblast growth factor 17. AB - Fibroblast growth factors are essential molecules for development. Here we characterize Fgfl7, a new member of the fibroblast growth factor (FGF) family. The Fgfl7 gene maps to mouse chromosome 14 and is highly conserved between mouse and human (93% identity). It exhibits 60% amino acid identity with Fgf8 and 50% identity with Fgf8. Both Fgf8 and Fgf17 have a similar structure and a similar pattern of alternative splicing in the 5' coding region. When expressed in 3T3 fibroblasts, mouse FGF17 is transforming, indicating that it can activate the 'c' splice form of either FGF receptor (FGFR) one or two. During midgestation embryogenesis, in situ hybridization analysis localized Fgf17 expression to specific sites in the midline structures of the forebrain, the midbrain-hindbrain junction, the developing skeleton and in developing arteries. Comparison to Fgf8 revealed a striking similarity in expression patterns, especially in the central nervous system (CNS), suggesting that both genes may be important for CNS development, although Fgf17 is expressed somewhat later than Fgf8. In the developing skeleton, both genes are expressed in costal cartilage while Fgf8 is preferentially expressed in long bones. In the developing great vessels Fgfl7 is preferentially expressed, suggesting that it may have a more prominent role in vascular growth. PMID- 10381578 TI - Expression of the medaka (Oryzias latipes) Ol-Rx3 paired-like gene in two diencephalic derivatives, the eye and the hypothalamus. AB - Here we report the expression pattern of the homeobox Ol-Rx3 gene, a medaka gene homologous to the mouse, Xenopus, zebrafish and Drosophila Rx genes. Ol-Rx3 starts to be expressed, at late gastrula stages, in the presumptive territories of the anterior brain. Subsequently, transcripts are localised in an antero ventral region of the prosencephalon and in the primordia of the optic vesicles. During organogenesis, distribution of Ol-Rx3 transcripts are gradually restricted to the floor of the diencephalon, the prospective territory of the hypothalamus and the neurohypophysis. During late development and in adult, Ol-Rx3 expression is maintained in hypothalamic nuclei bordering the third ventricle. In the optic vesicles, Ol-Rx3 expression is temporarily switched off when the eye cup morphogenesis is complete, but it is turned on again in the inner nuclear layer of the retina. Thus, the early expression pattern of Ol-Rx3 is in agreement with a conserved role in the specification of the ventral forebrain and eye field. Putative functions linked to late expression domains are discussed in light of the different hypothesis concerning the involvement of vertebrate Rx genes in the maintenance of particular cell fate. PMID- 10381579 TI - Expression of the optx2 homeobox gene during mouse development. AB - Optx2, a member of the sine oculis-Six family of homeobox genes, is first expressed in the anterior neural plate of the mouse embryo, and subsequently in the optic vesicle and ventral forebrain. During later development, expression is further restricted to precursors of the neural retina, optic chiasm, adenohypophysis and neurohypophysis. In the adult mouse retina, Optx2 mRNA is found in cells within the ganglion cell layer and inner nuclear layer. PMID- 10381580 TI - Expression of LKB1 and PTEN tumor suppressor genes during mouse embryonic development. AB - Germ-line mutations of LKB1 and PTEN tumor suppressor genes underlie the phenotypically related Peutz-Jeghers syndrome (PJS) and Cowden disease (CD), respectively. To analyze possible developmental roles of PTEN and LKB1, we have studied their mRNA expression during mouse embryonic development (E7-17.5) by in situ hybridization. Ubiquitous expression of both genes during early stages (E7 11) became more restricted in later embryonic development (E15-19) where LKB1 and PTEN showed prominent overlapping expression in e.g. gastrointestinal tract and lung. In contrast, LKB1 was selectively expressed at high levels in testis and PTEN was prominently expressed in skin epithelium and underlying mesenchyme. These results indicate that LKB1 and PTEN display largely overlapping expression patterns during embryonic development. Moreover, a high expression of these genes was observed in the tissues and organs affected in PJS and CD patients and in PTEN+/- mice. PMID- 10381581 TI - Topical review. Gastrin and gastric epithelial physiology. AB - Transepithelial transducing cells, particularly the gastrin (G) cell, co-ordinate gastric acid secretion with the arrival of food in the stomach. Recent work suggests that multiple active products are generated from the gastrin precursor, and that there are multiple control points in gastrin biosynthesis. Biosynthetic precursors and intermediates (progastrin and Gly-gastrins) are putative growth factors; their products, the amidated gastrins, regulate epithelial cell proliferation, the differentiation of acid-producing parietal cells and histamine secreting enterochromaffin-like (ECL) cells, and the expression of genes associated with histamine synthesis and storage in ECL cells, as well as acutely stimulating acid secretion. Gastrin also stimulates the production of members of the epidermal growth factor (EGF) family, which in turn inhibit parietal cell function but stimulate the growth of surface epithelial cells. Plasma gastrin concentrations are elevated in subjects with Helicobacter pylori, who are known to have increased risk of duodenal ulcer disease and gastric cancer. Studies of the physiology of gastrin may therefore contribute to an understanding of the mechanisms relevant to major upper gastrointestinal tract disease. PMID- 10381583 TI - Characterization of a new sodium channel mutation at arginine 1448 associated with moderate Paramyotonia congenita in humans. AB - 1. Paramyotonia congenita is a temperature-sensitive skeletal muscle disorder caused by missense mutations that occur in the adult skeletal muscle voltage gated sodium channel. We report here the identification of a new genetic mutation in a family with the paramyotonia congenita phenotype. 2. Single-strand conformation polymorphism analysis and DNA sequencing showed that the defect was linked to a single nucleotide substitution causing an amino acid change from an arginine to a serine at position 1448 in the human sodium channel alpha-subunit. 3. Expression of the altered protein in human embryonic kidney (HEK) 293 cells revealed several defects in channel function: (i) the rate of fast inactivation was slower in the mutant channel compared with wild-type, (ii) steady-state fast inactivation was shifted towards hyperpolarizing potentials, (iii) the R1448S channels deactivated much more slowly, and (iv) the mutant channels recovered from the fast inactivated state more rapidly. 4. By contrast, the activation curve, steady-state slow inactivation and the rate of onset and recovery from slow inactivation were not altered by the R1448S mutation. 5. These data show that the defects observed in the sodium channel function could well explain the onset of the paramyotonia congenita in this family and emphasize the role of segment S4 of domain IV in sodium channel inactivation. PMID- 10381582 TI - Involvement of the n-terminus of Kir6.2 in coupling to the sulphonylurea receptor. AB - 1. ATP-sensitive potassium (KATP) channels are composed of pore-forming Kir6.2 and regulatory SUR subunits. ATP inhibits the channel by interacting with Kir6.2, while sulphonylureas block channel activity by interaction with a high-affinity site on SUR1 and a low-affinity site on Kir6.2. MgADP and diazoxide interact with SUR1 to promote channel activity. 2. We examined the effect of N-terminal deletions of Kir6.2 on the channel open probability, ATP sensitivity and sulphonylurea sensitivity by recording macroscopic currents in membrane patches excised from Xenopus oocytes expressing wild-type or mutant Kir6.2/SUR1. 3. A 14 amino acid N-terminal deletion (DeltaN14) did not affect the gating, ATP sensitivity or tolbutamide block of a truncated isoform of Kir6.2, Kir6.2DeltaC26, expressed in the absence of SUR1. Thus, the N-terminal deletion does not alter the intrinsic properties of Kir6.2. 4. When Kir6.2DeltaN14 was coexpressed with SUR1, the resulting KATP channels had a higher open probability (Po = 0.7) and a lower ATP sensitivity (Ki = 196 microM) than wild-type (Kir6.2/SUR1) channels (Po = 0.32, Ki = 28 microM). High-affinity tolbutamide block was also abolished. 5. Truncation of five or nine amino acids from the N terminus of Kir6.2 also enhanced the open probability, and reduced both the ATP sensitivity and the fraction of high-affinity tolbutamide block, although to a lesser extent than for the DeltaN14 deletion. Site-directed mutagenesis suggests that hydrophobic residues in Kir6. 2 may be involved in this effect. 6. The reduced ATP sensitivity of Kir6.2DeltaN14 may be explained by the increased Po. However, when the Po was decreased (by ATP), tolbutamide was unable to block Kir6. 2DeltaN14/SUR1-K719A,K1385M currents, despite the fact that the drug inhibited Kir6.2-C166S/SUR1-K719A,K1385M currents (which in the absence of ATP have a Po of > 0.8 and are not blocked by tolbutamide). Thus the N-terminus of Kir6.2 may be involved in coupling sulphonylurea binding to SUR1 to closure of the Kir6.2 pore. PMID- 10381585 TI - Characterization of a C-type natriuretic peptide (CNP-39)-formed cation-selective channel from platypus (Ornithorhynchus anatinus) venom. AB - 1. The lipid bilayer technique is used to characterize the biophysical and pharmacological properties of a novel, fast, cation-selective channel formed by incorporating platypus (Ornithorhynchus anatinus) venom (OaV) into lipid membranes. 2. A synthetic C-type natriuretic peptide OaCNP-39, which is identical to that present in platypus venom, mimics the conductance, kinetics, selectivity and pharmacological properties of the OaV-formed fast cation-selective channel. The N-terminal fragment containing residues 1-17, i.e. OaCNP-39(1-17), induces the channel activity. 3. The current amplitude of the TEACl-insensitive fast cation-selective channel is dependent on cytoplasmic K+, [K+]cis. The increase in the current amplitude, as a function of increasing [K+]cis, is non-linear and can be described by the Michaelis-Menten equation. At +140 mV, the values of gammamax and KS are 63.1 pS and 169 mM, respectively, whereas at 0 mV the values of gammamax and KS are 21.1 pS and 307 mM, respectively. gammamax and KS are maximal single channel conductance and concentration for half-maximal gamma, respectively. The calculated permeability ratios, PK:PRb:PNa:PCs:PLi, were 1:0.76:0.21:0.09:0.03, respectively. 4. The probability of the fast channel being open, Po, increases from 0.15 at 0 mV to 0.75 at +140 mV. In contrast, the channel frequency, Fo, decreases from 400 to 180 events per second for voltages between 0 mV and +140. The mean open time, To, increases as the bilayer is made more positive, between 0 and +140 mV. The mean values of the voltage-dependent kinetic parameters, Po, Fo, To and mean closed time (Tc), are independent of [KCl]cis between 50 and 750 mM (P > 0. 05). 5. It is proposed that some of the symptoms of envenomation by platypus venom may be caused partly by changes in cellular functions mediated via the OaCNP-39-formed fast cation-selective channel, which affects signal transduction. PMID- 10381584 TI - Properties of heterologously expressed hTRP3 channels in bovine pulmonary artery endothelial cells. AB - 1. We combined patch clamp and fura-2 fluorescence methods to characterize human TRP3 (hTRP3) channels heterologously expressed in cultured bovine pulmonary artery endothelial (CPAE) cells, which do not express the bovine trp3 isoform (btrp3) but express btrp1 and btrp4. 2. ATP, bradykinin and intracellular InsP3 activated a non-selective cation current (IhTRP3) in htrp3-transfected CPAE cells but not in non-transfected wild-type cells. During agonist stimulation, the sustained rise in [Ca2+]i was significantly higher in htrp3-transfected cells than in control CPAE cells. 3. The permeability for monovalent cations was PNa > PCs approximately PK >> PNMDG and the ratio PCa/PNa was 1.62 +/- 0.27 (n = 11). Removal of extracellular Ca2+ enhanced the amplitude of the agonist-activated IhTRP3 as well as that of the basal current The trivalent cations La3+ and Gd3+ were potent blockers of IhTRP3 (the IC50 for La3+ was 24.4 +/- 0.7 microM). 4. The single-channel conductance of the channels activated by ATP, assessed by noise analysis, was 23 pS. 5. Thapsigargin and 2,5-di-tert-butyl-1, 4 benzohydroquinone (BHQ), inhibitors of the organellar Ca2+-ATPase, failed to activate IhTRP3. U-73122, a phospholipase C blocker, inhibited IhTRP3 that had been activated by ATP and bradykinin. Thimerosal, an InsP3 receptor-sensitizing compound, enhanced IhTRP3, but calmidazolium, a calmodulin antagonist, did not affect IhTRP3. 6. It is concluded that hTRP3 forms non-selective plasmalemmal cation channels that function as a pathway for agonist-induced Ca2+ influx. PMID- 10381586 TI - Modulation of rat cardiac sodium channel by the stimulatory G protein alpha subunit. AB - 1. Modulation of cardiac sodium currents (INa) by the G protein stimulatory alpha subunit (Gsalpha) was studied using patch-clamp techniques on freshly dissociated rat ventricular myocytes. 2. Whole-cell recordings showed that stimulation of beta-adrenergic receptors with 10 microM isoprenaline (isoproterenol, ISO) enhanced INa by 68.4 +/- 9.6 % (mean +/- s.e.m.; n = 7, P < 0.05 vs. baseline). With the addition of 22 microgram ml-1 protein kinase A inhibitor (PKI) to the pipette solution, 10 microM ISO enhanced INa by 30.5 +/- 7.0 % (n = 7, P < 0.05 vs. baseline). With the pipette solution containing both PKI and 20 microgram ml 1 anti-Gsalpha IgG or 20 microgram ml-1 anti-Gsalpha IgG alone, 10 microM ISO produced no change in INa. 3. The effect of Gsalpha on INa was not due to changes in the steady-state activation or inactivation curves, the time course of current decay, the development of inactivation, or the recovery from inactivation. 4. Whole-cell INa was increased by 45.2 +/- 5.3% (n = 13, P < 0.05 vs. control) with pipette solution containing 1 microM Gsalpha27-42 peptide (amino acids 27-42 of rat brain Gsalpha) without altering the properties of Na+ channel kinetics. Furthermore, application of 1 nM Gsalpha27-42 to Na+ channels in inside-out macropatches increased the ensemble-averaged INa by 32.5 +/- 6.8 % (n = 8, P < 0.05 vs. baseline). The increase in INa was reversible upon Gsalpha27-42 peptide washout. Single channel experiments showed that the Gsalpha27-42 peptide did not alter the Na+ single channel current amplitude, the mean open time or the mean closed time, but increased the number of functional channels (N) in the patch. 5. Application of selected short amino acid segments (Gsalpha27-36, Gsalpha33-42 and Gsalpha30-39) of the 16 amino acid Gsalpha peptide (Gsalpha27-42 peptide) showed that only the C-terminal segment of this peptide (Gsalpha33-42) significantly increased INa in a dose-dependent fashion. These results show that cardiac INa is regulated by Gsalpha via a mechanism independent of PKA that results in an increase in the number of functional Na+ channels. In addition, a 10 residue domain (amino acids 33-42) near the N-terminus of Gsalpha is important in modulating cardiac Na+ channels. PMID- 10381587 TI - Evidence for phosphorylation-dependent internalization of recombinant human rho1 GABAC receptors. AB - 1. Recombinant wild-type or mutant human rho1 GABA receptors were expressed in human embryonic kidney (HEK) 293 or monkey COS-7 cells and studied using the patch clamp technique. 2. Standard whole-cell recordings with 4 mM Mg-ATP in the patch pipette induced a time-dependent decrease in the GABA-activated current (IGABA) amplitude that was not the result of a decrease in GABA sensitivity. In contrast, IGABA remained stable when recordings were obtained using the perforated patch configuration or with standard whole-cell recording and no Mg ATP in the patch pipette. 3. The inhibitors of serine/threonine protein kinases KN-62 (20 microM) or staurosporine (20 nM) prevented the time-dependent decrease in the amplitude of IGABA seen in the presence of ATP. Alkaline phosphatase (220 U ml-1), when added to the patch pipette in the absence of ATP, induced a transient potentiation of IGABA. Although the protein kinase C (PKC) activator 4beta-phorbol 12-myristate, 13-acetate (PMA) did not reduce the amplitude of IGABA, inclusion of the catalytic domain of PKC in the recording pipette accelerated the time-dependent decrease in current amplitude. These data suggest that phosphorylation is involved in the regulation of the amplitude of IGABA. 4. Mutation of the three PKC consensus sequences of the rho1 receptor had no significant effect on the decline in IGABA, indicating that direct phosphorylation of these putative sites on the rho1 receptor does not underlie the time-dependent decrease in amplitude. 5. In COS-7 cells transfected with wild type rho1 receptors, the amplitude of IGABA had completely recovered to the original value when the same cells were repatched after 30-40 min, indicating that the decline in IGABA was a reversible process. 6. The inhibitor of actin filament formation cytochalasin B, when added to the patch pipette in the absence of ATP, induced a time-dependent inactivation suggesting that the actin cytoskeleton may play a role in the regulation of the amplitude. 7. Coincident with the decrease in the amplitude of IGABA, the cell capacitance significantly decreased in the presence of ATP in the patch pipette. This decrease in capacitance was not observed in the absence of Mg-ATP. The decrease in the membrane surface area suggests that receptor internalization could be a potential mechanism for the observed inactivation. 8. At 32 C, compared with 22 C, the rate and magnitude of the decline was increased dramatically. In contrast, at 16 C, no significant change in IGABA was observed over the 20 min recording time. This marked temperature sensitivity is consistent with receptor internalization as a mechanism for the time-dependent decline in IGABA. 9. The specificity of the decrease in IGABA was assessed by coexpressing the voltage-dependent potassium channel Kv1.4 along with the rho1 receptor in HEK293 cells. The amplitude of the potassium current (IKv1.4) exhibited very little decrement in comparison to IGABA suggesting that the putative GABA receptor internalization was not the consequence of a non-specific membrane retrieval. PMID- 10381588 TI - The erg-like potassium current in rat lactotrophs. AB - 1. The ether-a-go-go-related gene (erg)-like K+ current in rat lactotrophs from primary culture was characterized and compared with that in clonal rat pituitary cells (GH3/B6). The class III antiarrhythmic E-4031 known to block specifically erg K+ channels was used to isolate the erg-like current as the E-4031-sensitive current. The experiments were performed in 150 mM K+ external solution using the patch-clamp technique. 2. The erg-like K+ current elicited with hyperpolarizing pulses negative to -100 mV consisted of a fast and a pronounced slowly deactivating current component. The contribution of the slow component to the total current amplitude was potential dependent and varied from cell to cell. At 100 mV it ranged from 50 to 85% and at -140 mV from 21 to 45%. 3. The potential dependent channel availability curves determined with 2 s prepulses were fitted with the sum of two Boltzmann functions. The function related to the slowly deactivating component of the erg-like current was shifted by more than 40 mV to more negative membrane potentials compared with that of the fast component. 4. In contrast to that of native lactotrophs studied under identical conditions, the erg-like K+ current of GH3/B6 cells was characterized by a predominant fast deactivating current component, with similar kinetic and steady-state properties to the fast deactivating current component of native lactotrophs. 5. Thyrotrophin releasing hormone reduced the erg-like current in native lactotrophs via an intracellular signal cascade which seemed to involve a pathway independent from protein kinase A and protein kinase C. 6. RT-PCR studies on cytoplasm from single lactotrophs revealed the presence of mRNA of the rat homologue of the human ether a-go-go-related gene HERG (r-erg1) as well as mRNA of the two other cloned r-erg cDNAs (r-erg2 and r-erg3) in different combinations. In GH3/B6 cells, only the transcripts of r-erg1 and r-erg2 were found. PMID- 10381589 TI - Activation of the Na+-K+ pump by hyposmolality through tyrosine kinase-dependent Cl- conductance in Xenopus renal epithelial A6 cells. AB - 1. We studied the regulatory mechanism of Na+ transport by hyposmolality in renal epithelial A6 cells. 2. Hyposmolality increased (1) Na+ absorption, which was detected as an amiloride-sensitive short-circuit current (INa), (2) Na+-K+ pump activity, (3) basolateral Cl- conductance (Gb,Cl), and (4) phosphorylation of tyrosine, suggesting an increase in activity of protein tyrosine kinase (PTK). 3. A Cl- channel blocker, 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB), which abolished Gb, Cl, blocked the INa by inhibiting the Na+-K+ pump without any direct effect on amiloride-sensitive Na+ channels. Diminution of Gb,Cl by Cl- replacement with a less permeable anion, gluconate, also decreased the hyposmolality-increased Na+-K+ pump activity. 4. The PTK inhibitors tyrphostin A23 and genistein induced diminution of the hyposmolality-stimulated Gb,Cl, which was associated with attenuation of the hyposmolality-increased Na+-K+ pump activity. 5. Taken together, these observations suggest that: (1) hyposmolality activates PTK; (2) the activated PTK increases Gb,Cl; and (3) the PTK-increased Gb,Cl stimulates the Na+-K+ pump. 6. This PTK-activated Gb,Cl-mediated signalling of hyposmolality is a novel pathway for stimulation of the Na+-K+ pump. PMID- 10381590 TI - Twitch-potentiation increases calcium in peripheral more than in central mitochondria of guinea-pig ventricular myocytes. AB - 1. The mitochondrial total calcium content ([Ca]mt) was studied with electron probe microanalysis (EPMA) in isolated guinea-pig ventricular myocytes in order to answer the question of whether electrical stimulation increases [Ca]mt in subsarcolemmal and central mitochondria to a different extent. 2. In unstimulated myocytes subsarcolemmal [Ca]mt was (mean +/- s.e.m.) 535 +/- 229 micromol (kg dry weight (DW))-1 and central [Ca]mt was 513 +/- 162 micromol (kg DW)-1. These values do not differ and correspond to approximately 180 micromol calcium per litre of mitochondria or 180 microM. 3. Contractions were potentiated to an optimum by stimulation with trains of 12 paired stimuli. After potentiation with 12 paired action potentials, cells were shock-frozen 120 ms after the start of the first action potential of the 13th pair. Subsarcolemmal [Ca]mt was 1.3 +/- 0.4 mmol (kg DW)-1 (433 microM) and central [Ca]mt was 227 +/- 104 micromol (kg DW)-1 (76 microM). The difference was significant. 4. After potentiation with 12 paired voltage-clamp pulses, cells were shock-frozen 120 ms after the start of the first pulse of the 13th pair. Subsarcolemmal [Ca]mt was 2.2 +/- 1.0 mmol (kg DW)-1 (733 microM) and central [Ca]mt was 630 +/- 180 micromol (kg DW)-1 (210 microM). After removal of extracellular K+, five paired voltage-clamp pulses increased subsarcolemmal [Ca]mt to 2.1 +/- 0.8 mmol (kg DW)-1 (700 microM), which was significantly higher than the central [Ca]mt of 389 +/- 88 micromol (kg DW) 1 or 130 microM. 5. In unstimulated cells, [Na] and [K] in subsarcolemmal and central mitochondria were not different. In potentiated myocytes, subsarcolemmal [Na]mt was 236 +/- 20 mmol (kg DW)-1 or 79 mM, which is significantly higher than the central [Na]mt of 50 +/- 5 mmol (kg DW)-1 or 16 mM. 6. The differences in [Ca]mt and [Na]mt are attributed to subsarcolemmal cytosolic microdomains of elevated [Ca2+] and [Na+] generated during contractile potentiation by transmembrane Ca2+ and Na+ fluxes. PMID- 10381592 TI - Calcium binding capacity of the cytosol and endoplasmic reticulum of mouse pancreatic acinar cells. AB - 1. The droplet technique was used in this study to measure total calcium loss from pancreatic acinar cells due to calcium extrusion. The calcium binding capacity of the cytosol (kc) was measured as the ratio of the decrease in the total calcium concentration of the cytosol of the cell (Delta[Ca]c) and the synchronously occurring decrease in the free calcium ion concentration in the cytosol (Delta[Ca2+]c). The calcium dependency of the calcium binding capacity was determined by plotting values of kc against the corresponding [Ca2+]c. The rise in the cytosolic Ca2+ concentration of pancreatic acinar cells was triggered by stimulation with a supramaximal dose of cholecystokinin (CCK). The recovery of [Ca2+]c during continued exposure to the agonist was due to calcium extrusion from the cell. 2. The calcium binding capacity was about 1500-2000 for the [Ca2+]c range 150-500 nM. The mechanism of buffering was not investigated in this study. The calcium binding capacity of the cytosol did not vary significantly with [Ca2+]c in this range. The CCK-evoked decrease in the total calcium concentration in the lumen of the endoplasmic reticulum (ER) can be estimated from our data, taking into account previously published values for the volume of the ER in pancreatic acinar cells. Comparing the decrease in the total ER calcium concentration with our recently reported values for agonist-induced reductions in the free Ca2+ concentration inside the ER, we estimate that the calcium binding capacity of the ER is approximately 20. In pancreatic acinar cells we have therefore found a difference of two orders of magnitude in the efficiency of calcium buffering in the cytosol and the ER lumen. PMID- 10381591 TI - Positive and negative inotropic effects of NO donors in atrial and ventricular fibres of the frog heart. AB - 1. The cardiac effects of the NO donors sodium nitroprusside (SNP), S-nitroso-N acetyl-penicillamine (SNAP) and 3-morpholino-sydnonimine (SIN-1) were studied in frog fibres to evaluate the contribution of cyclic GMP-dependent mechanisms. 2. SNP and SNAP (0.1-100 microM) reduced the force of contraction in a concentration dependent manner in atrial and ventricular fibres. This effect was associated with a reduction in the time to peak (TTP) and the time for half-relaxation of contraction (T). 3. SIN-1 (100 microM) also reduced the force of contraction in two-thirds of the atrial fibres. However, it exerted a positive inotropic effect in the remaining atrial fibres, as well as in most ventricular fibres. 4. The guanylyl cyclase inhibitor 1H-[1,2,4]oxidiazolo[4,3-a]quinoxaline-1-one (ODQ, 10 microM) antagonized the negative inotropic effects of SIN-1 (50 microM) and SNAP (25 microM) but had no effect on the positive inotropic response to SIN-1 (100 microM). 5. In the presence of SIN-1, superoxide dismutase (SOD, 50-200 U ml-1) either potentiated the negative inotropic effect or turned the positive inotropic effect of the drug into a negative effect. SOD had no effects when applied alone or in the presence of SNAP. 6. 6-Anilino-5,8-quinolinedione (LY 83583, 3-30 microM), a superoxide anion generator also known as a cyclic GMP-lowering agent, exerted a positive inotropic effect, which was antagonized by SOD (200-370 U ml 1) but not by ODQ (10 microM). 7. We conclude that SNP, SNAP and SIN-1 exert cyclic GMP-dependent negative inotropic effects, which are attributed to the generation of NO. In addition, SIN-1 and LY 83583 exert cyclic GMP-independent positive inotropic effects, which require the generation of superoxide anion. PMID- 10381593 TI - Properties of Ca2+ sparks evoked by action potentials in mouse ventricular myocytes. AB - 1. Calcium sparks were examined in enzymatically dissociated mouse cardiac ventricular cells using the calcium indicator fluo-3 and confocal microscopy. The properties of the mouse cardiac calcium spark are generally similar to those reported for other species. 2. Examination of the temporal relationship between the action potential and the time course of calcium spark production showed that calcium sparks are more likely to occur during the initial repolarization phase of the action potential. The latency of their occurrence varied by less than 1.4 ms (s.d.) and this low variability may be explained by the interaction of the gating of L-type calcium channels with the changes in driving force for calcium entry during the action potential. 3. When fixed sites within the cell are examined, calcium sparks have relatively constant amplitude but the amplitude of the sparks was variable among sites. The low variability of the amplitude of the calcium sparks suggests that more than one sarcoplasmic reticulum (SR) release channel must be involved in their genesis. Noise analysis (with the assumption of independent gating) suggests that > 18 SR calcium release channels may be involved in the generation of the calcium spark. At a fixed site, the response is close to 'all-or-none' behaviour which suggests that calcium sparks are indeed elementary events underlying cardiac excitation-contraction coupling. 4. A method for selecting spark sites for signal averaging is presented which allows the time course of the spark to be examined with high temporal and spatial resolution. Using this method we show the development of the calcium spark at high signal-to noise levels. PMID- 10381594 TI - Light adaptation and dark adaptation of human rod photoreceptors measured from the a-wave of the electroretinogram. AB - 1. We recorded the a-wave of the human electroretinogram from subjects with normal vision, using a corneal electrode and ganzfeld (full-field) light stimulation. From analysis of the rising phase of rod-isolated flash responses we determined the maximum size (amax) of the a-wave, a measure of the massed circulating current of the rods, and the amplification constant (A) of transduction within the rod photoreceptors. 2. During light adaptation by steady backgrounds the maximal response was reduced, as reported previously. amax declined approximately as I0/(I0 + IB), where IB is retinal illuminance and I0 is a constant. In different subjects I0 ranged from 40 to 100 trolands, with a mean of 70 trolands, corresponding to about 600 photoisomerizations s-1 per rod. (1 troland is the retinal illuminance that results when a surface luminance of 1 cd m-2 is viewed through a pupil area of 1 mm2.) The amplification constant A decreased only slightly in the presence of steady backgrounds. 3. Following a full bleach amax recovered along an S-shaped curve over a period of 30 min. There was no detectable response for the first 5 min, and half-maximal recovery took 13 17 min. 4. The apparent amplification constant decreased at early times after large bleaches. However, upon correction for reduced light absorption due to loss of pigment, with regeneration of rhodopsin occurring with a time constant of 9-15 min in different subjects, it appeared that the true value of A was probably unchanged by bleaching. 5. The recovery of amax following a bleach could be converted into recovery of equivalent background intensity, using a 'Crawford transformation' derived from the light adaptation results. Following bleaches ranging from 10 to > 99 %, the equivalent background intensity decayed approximately exponentially, with a time constant of about 3 min. 6. The time taken for amax to recover to a fixed proportion of its original level increased approximately linearly (rather than logarithmically) with fractional bleach, with a slope of about 12 min per 100 % bleach. Similar behaviour has previously been seen in psychophysical dark adaptation experiments, for the dependence of the 'second component' of recovery on the level of bleaching. PMID- 10381595 TI - Dual mechanism for presynaptic modulation by axonal metabotropic glutamate receptor at the mouse mossy fibre-CA3 synapse. AB - 1. To investigate mechanisms responsible for the presynaptic inhibitory action mediated by the axonal group II metabotropic glutamate receptor (mGluR) at the mossy fibre-CA3 synapse, we used a quantitative fluorescence measurement of presynaptic Ca2+ in mouse hippocampal slices. 2. Bath application of the group II mGluR-specific agonist (2S,1'R,2'R,3'R)-2-(2, 3-dicarboxycyclopropyl)glycine (DCG IV, 1 microM) reversibly suppressed the presynaptic Ca2+ influx (to 55.2 +/- 4.6 % of control, n = 5) as well as field EPSPs recorded simultaneously (to 3.1 +/- 2.0%). Presynaptic fibre volley was not affected by 1 microM DCG-IV. 3. A quantitative analysis of the inhibition of presynaptic Ca2+ influx and field EPSP suggested that DCG-IV suppressed the field EPSP to a greater extent than would be expected if the suppression were solely due to a decrease in the presynaptic Ca2+ influx. 4. DCG-IV at 1 microM suppressed the mean frequency (to 73.8 +/- 3.9% of control, n = 11), but not the mean amplitude (to 97.0 +/- 3.5%), of miniature EPSCs recorded from CA3 neurones using the whole-cell patch-clamp technique. 5. These results suggest that group II mGluR-mediated suppression is due both to a reduction of presynaptic Ca2+ influx and downregulation of the subsequent exocytotic machinery. PMID- 10381596 TI - Serotonergic modulation of hyperpolarization-activated current in acutely isolated rat dorsal root ganglion neurons. AB - 1. The effect of serotonin (5-HT) on the hyperpolarization-activated cation current (IH) was studied in small-, medium- and large-diameter acutely isolated rat dorsal root ganglion (DRG) cells, including cells categorized as type 1, 2, 3 and 4 based on membrane properties. 5-HT increased IH in 91 % of medium-diameter DRG cells (including type 4) and in 67 % of large-diameter DRG cells, but not other DRG cell types. 2. The increase of IH by 5-HT was antagonized by spiperone but not cyanopindolol, and was mimicked by 5-carboxyamidotryptamine, but not (+) 8-hydroxydipropylaminotetralin (8-OH-DPAT) or cyanopindolol. These data suggested the involvement of 5-HT7 receptors, which were shown to be expressed by medium diameter DRG cells using RT-PCR analysis. 3. 5-HT shifted the conductance-voltage relationship of IH by +6 mV without changing peak conductance. The effects of 5 HT on IH were mimicked and occluded by forskolin, but not by inactive 1,9-dideoxy forskolin. 4. At holding potentials negative to -50 mV, 5-HT increased steady state inward current and instantaneous membrane conductance (fast current). The 5 HT-induced inward current and fast current were blocked by Cs+ but not Ba2+ and reversed at -23 mV, consistent with the properties of tonically activated IH. 5. In medium-diameter neurons recorded from in the current clamp mode, 5-HT depolarized the resting membrane potential, decreased input resistance and facilitated action potential generation by anode-break excitation. 6. The above data suggest that in distinct subpopulations of DRG neurons, 5-HT increases cAMP levels via activation of 5-HT7 receptors, which shifts the voltage dependence of IH to more depolarized potentials and increases neuronal excitability. PMID- 10381597 TI - Presynaptic serotonergic inhibition of GABAergic synaptic transmission in mechanically dissociated rat basolateral amygdala neurons. AB - 1. The basolateral amygdala (ABL) nuclei contribute to the process of anxiety. GABAergic transmission is critical in these nuclei and serotonergic inputs from dorsal raphe nuclei also significantly regulate GABA release. In mechanically dissociated rat ABL neurons, spontaneous miniature inhibitory postsynaptic currents (mIPSCs) arising from attached GABAergic presynaptic nerve terminals were recorded with the nystatin-perforated patch method and pharmacological isolation. 2. 5-HT reversibly reduced the GABAergic mIPSC frequency without affecting the mean amplitude. The serotonergic effect was mimicked by the 5-HT1A specific agonist 8-OH DPAT (8-hydroxy-2-(di-n-propylamino)tetralin) and blocked by the 5-HT1A antagonist spiperone. 3. The GTP-binding protein inhibitor N ethylmaleimide removed the serotonergic inhibition of mIPSC frequency. In either K+-free or Ca2+-free external solution, 5-HT could inhibit mIPSC frequency. 4. High K+ stimulation increased mIPSC frequency and 8-OH DPAT inhibited this increase even in the presence of Cd2+. 5. Forskolin, an activator of adenylyl cyclase (AC), significantly increased synaptic GABA release frequency. Pretreatment with forskolin prevented the serotonergic inhibition of mIPSC frequency in both the standard and high K+ external solution. 6. Ruthenium Red (RR), an agent facilitating the secretory process in a Ca2+-independent manner, increased synaptic GABA release. 5-HT also suppressed RR-facilitated mIPSC frequency. 7. We conclude that 5-HT inhibits GABAergic mIPSCs by inactivating the AC-cAMP signal transduction pathway via a G-protein-coupled 5-HT1A receptor and this intracellular pathway directly acts on the GABA-releasing process independent of K+ and Ca2+ channels in the presynaptic nerve terminals. PMID- 10381598 TI - Ca2+ permeability and kinetics of glutamate receptors in rat medial habenula neurones: implications for purinergic transmission in this nucleus. AB - 1. We have previously investigated P2X receptor-mediated synaptic currents in medial habenula neurones and shown that they can be calcium permeable. We now investigate the receptor properties of glutamate, the other, more abundant excitatory transmitter, to determine its receptor subtypes and their relative calcium permeability. This may have implications for the physiological role of the P2X receptors which mediate synaptic currents. 2. Using fast application of ATP, L-glutamate or kainate to nucleated patches, glutamate receptors were determined to be of the AMPA subtype but no functional P2X receptors were detected. 3. The deactivation and desensitization rates of the AMPA channel were determined to have time constants of 1.77 +/- 0.21 ms (n = 10) and 4.01 +/- 0.85 ms (n = 9) at -60 mV, respectively. AMPA receptors recovered from desensitization with two exponential components with time constants of 21.08 +/- 2.95 and 233.60 +/- 51.1 ms (n = 3). None of the deactivation or desensitization properties of the GluR channels depended on membrane potential. 4. The current-voltage relationship under different ionic conditions revealed that the GluR channel was equally permeable to Cs+ and Na+ but relatively impermeable to Ca2+ (PCa/PCs = 0.13, n = 6). 5. For both synaptic currents and somatic currents activated by fast application of L-glutamate to nucleated patches, decay time constants were similar at +/-60 mV in the presence of Mg2+ ions. Thus GluR channels appear to be of the AMPA subtype and not the NMDA subtype. 6. Thus, under the conditions of this study, neurones of the medial habenula lack functional NMDA receptors and possess AMPA receptors that have low permeability to Ca2+. We conclude that the P2X receptor-mediated synaptic currents are the only calcium-permeable fast transmitter gated currents in these neurones which may be important for their physiological function. PMID- 10381599 TI - Mechanism underlying increased neuronal activity in the rat ventrolateral periaqueductal grey by a mu-opioid. AB - 1. The overall effect of the mu-opioid receptor agonist DAMGO (Tyr-D-Ala-Gly MePhe-Gly-ol) on ventrolateral periaqueductal grey (PAG) neurons in brain slices was studied using the whole-cell patch-clamp recording technique. 2. Under current-clamp conditions, DAMGO (1 microM) increased cell firing in many PAG neurons even though the opioid induced hyperpolarization and inhibited excitatory postsynaptic potentials (EPSPs) in these cells. 3. The increase in cell activity by DAMGO was observed in both transverse and horizontal slices. The increase persisted when the membrane potential was re-depolarized to the control level. Thus, different planes of sections or the removal of Na+ channel inactivation could not account for the observation. 4. The GABA antagonist bicuculline caused cell firing, mimicking the excitatory effect of DAMGO. Unlike DAMGO, however, bicuculline depolarized PAG cells. 5. Under voltage-clamp conditions, with the same driving force, the evoked inhibitory postsynaptic currents (IPSCs) in these neurons were 2.3 times larger than the evoked excitatory postsynaptic currents (EPSCs). Furthermore, DAMGO inhibited IPSCs by 60.7% while it inhibited EPSCs by 35.3%. 6. We propose that the overall effect of an opioid depends on the dynamic balance of its excitatory and inhibitory actions. In the PAG, the blockade of the inhibitory drive of GABAergic inputs by DAMGO is large. It overcomes the DAMGO induced hyperpolarization and inhibition of EPSCs and thus results in the excitation of these neurons. PMID- 10381601 TI - Rapid report: postsynaptic bursting is essential for 'Hebbian' induction of associative long-term potentiation at excitatory synapses in rat hippocampus. AB - 1. The biologically relevant rules of synaptic potentiation were investigated in hippocampal slices from adult rat by mimicking neuronal activity seen during learning behaviours. Synaptic efficacy was monitored in two separate afferent pathways among the Schaffer collaterals during intracellular recording of CA1 pyramidal neurones. The effects of pairing presynaptic single spikes or bursts with postsynaptic single spikes or bursts, repeated at 5 Hz ('theta' frequency), were compared. 2. The pairing of ten single evoked excitatory synaptic events with ten postsynaptic single action potentials at 5 Hz, repeated twelve times, failed to induce synaptic enhancement (EPSP amplitude 95% of baseline amplitude 20 min after pairing; n = 5). In contrast, pairing the same number of action potentials, but clustered in bursts, induced robust synaptic potentiation (EPSP amplitude 163%; P < 0.01, Student's t test; n = 5). This potentiation was input specific, long lasting ( > 1 h; n = 3) and its induction was blocked by an antagonist at NMDA receptors (20-50 microM D(-)-2-amino-5-phosphonopentanoic acid; EPSP amplitude 109%; n = 6). 3. Presynaptic bursting paired with postsynaptic single action potentials did not induce input specific synaptic change (113 % in the test input vs. 111 % in the control; n = 8). In contrast, postsynaptic bursting when paired with presynaptic single action potentials was sufficient to induce synaptic potentiation when the presynaptic activity preceded the postsynaptic activity by 10 ms (150 vs. 84 % in the control input; P < 0.01; n = 10). 4. These results indicate that, under our conditions, postsynaptic bursting activity is necessary for associative synaptic potentiation at CA1 excitatory synapses in adult hippocampus. The existence of a distinct postsynaptic signal for induction of synaptic change calls for refinement of the common interpretation of Hebb's rule, and is likely to have important implications for our understanding of cortical network operation. PMID- 10381600 TI - Tumour necrosis factor-alpha activates a calcium sensitization pathway in guinea pig bronchial smooth muscle. AB - 1. The effects of tumour necrosis factor-alpha (TNF) on guinea-pig bronchial smooth muscle contractility were investigated. 2. The Ca2+-activated contractile response of permeabilized bronchial smooth muscle strips was significantly increased after incubation with 1 microgram ml-1 TNF for 45 min. This TNF-induced effect was not due to a further increase in intracellular Ca2+. 3. The TNF induced Ca2+ sensitization was, at least partly, the result of an increase in myosin light chain20 phosphorylation. 4. The intracellular signalling pathway involved in this effect of TNF was further investigated. Sphingomyelinase, a potential mediator of TNF, had no effect on Ca2+ sensitivity of permeabilized bronchial smooth muscle. Also, p42/p44 mitogen-activated protein kinase (p42/p44mapk), activated by TNF in some cell types, did not show an increased activation in bronchial smooth muscle after TNF treatment. 5. In conclusion, TNF may activate a novel signalling pathway in guinea-pig bronchial smooth muscle leading to an increase in myosin light chain20 phosphorylation and a subsequent increase in Ca2+ sensitivity of the myofilaments. This pathway does not appear to involve sphingomyelinase-liberated ceramides or activation of p42/p44mapk. Given the importance of TNF in asthma, this TNF-induced Ca2+ sensitization of the myofilaments may represent a mechanism responsible for airway hyper responsiveness. PMID- 10381602 TI - Rapid report: a novel technique for quantitative measurement of free Ca2+ concentration in rat heart mitochondria. AB - 1. The free mitochondrial Ca2+ concentration ([Ca2+]m) in rat heart mitochondria was measured quantitatively by loading the mitochondria with fura-2 and then injecting them into Xenopus laevis oocytes. 2. When oocytes were incubated with a physiological solution, the free cytosolic Ca2+ concentration ([Ca2+]c) in the oocytes was 82 +/- 11 nM (n = 20, mean +/- s.e.m.) and the [Ca2+]m of the injected rat heart mitochondria was 116 +/- 10 nM (n = 18, mean +/- s.e.m.). 3. Inhibition of the oocyte endoplasmic reticular Ca2+-ATPase with thapsigargin produced a transient increase in averaged [Ca2+]c at sub-micromolar concentrations. 4. Injection of cardiac mitochondria blunted the peak and prolonged the duration of thapsigargin-induced [Ca2+]c transients as a result of Ca2+ sequestration by the cardiac mitochondria. 5. These results demonstrate that the present technique provides a new approach for studying [Ca2+]m regulation quantitatively under physiological environments. Furthermore, it clearly shows that cardiac mitochondria can modify the shape of thapsigargin-induced cytosolic Ca2+ pulses in Xenopus oocytes. PMID- 10381604 TI - Rhythmic neuronal activity in the lateral cerebellum of the cat during visually guided stepping. AB - 1. The discharge patterns of 117 lateral cerebellar neurones were studied in cats during visually guided stepping on a horizontal circular ladder. Ninety per cent of both nuclear cells (53/59) and Purkinje cells (53/58) showed step-related rhythmic modulations of their discharge frequency (one or more periods of 'raised activity' per step cycle of the ipsilateral forelimb). 2. For 31% of nuclear cells (18/59) and 34% of Purkinje cells (20/58) the difference between the highest and lowest discharge rates in different parts of the step cycle was > 50 impulses s-1. 3. Individual neurones differed widely in the phasing of their discharges relative to the step cycle. Nevertheless, for both Purkinje cells and nuclear cells population activity was significantly greater in swing than in stance; the difference was more marked for the nuclear population. 4. Some cells exhibited both step-related rhythmicity and visual responsiveness (28 of 67 tested, 42%), whilst others were rhythmically active during locomotion and increased their discharge rate ahead of saccadic eye movements (11 of 54 tested, 20%). The rhythmicity of cells that were visually responsive was typical of the rhythmicity seen in the whole locomotor-related population. The step-related rhythmicity of cells that also discharged in relation to saccades was generally below average strength compared with the cortical and nuclear populations as a whole. 5. The possibility is discussed that the rhythmicity of dentate neurones acts as a powerful source of excitatory locomotor drive to motor cortex, and may thereby contribute to establishing the step-related rhythmicity of motor cortical (including pyramidal tract) neurones. More generally, the activity patterns of lateral cerebellar neurones provide for a role in the production of visually guided, co-ordinated eye and body movements. PMID- 10381603 TI - Brain sites of action of endogenous interleukin-1 in the febrile response to localized inflammation in the rat. AB - 1. Interleukin (IL)-1 is a potent endogenous pyrogen which causes fever when injected into a number of brain sites. However, the brain sites at which endogenous IL-1 acts to influence body temperature remain equivocal. The aim of this study was to determine the effect of local administration of the interleukin 1 receptor antagonist (IL-1ra) into specific sites in the hypothalamus, and other brain regions known to contain receptors for IL-1, on the febrile response of rats to peripheral injection of lipopolysaccharide (LPS) into a subcutaneous air pouch (intrapouch, i.p.o.) that does not lead to LPS appearance in the circulation. 2. Injection of LPS (100 microgram kg-1, i.p.o.) induced a rise in body temperature which commenced 1.5 h after injection and was maximal at 3 h (38.9 +/- 0.2 C, compared with 37.0 +/- 0.1 C at 0 h, n = 6, P < 0.001). Intracerebroventricular (i.c.v.) IL-1ra (500 microgram in 5 microliter) significantly attenuated LPS fever (IL-1ra, 37.7 +/- 0.2 C; saline, 38.9 +/- 0.2 C; n = 6, P < 0.001). Unilateral microinjection of IL-1ra (50 microgram in 0.5 microliter at 0 + 1 h) into the anterior hypothalamus (AH), paraventricular hypothalamic nucleus (PVH), peri-subfornical organ, subfornical organ (SFO) or hippocampus (dentate gyrus and CA3 region) also significantly reduced the fever induced by LPS. 3. The same dose of IL-1ra had no effect on fever when administered into the ventromedial hypothalamus (VMH), organum vasculosum lamina terminalis (OVLT), CA1 field of the hippocampus, striatum or cortex. 4. These data indicate that the action of endogenous IL-1 in the brain during fever is site specific, acting at the AH, PVH, SFO and hippocampus, but not the VMH, OVLT and striatum or cortex. PMID- 10381605 TI - Non-chemical inhibition of respiratory motor output during mechanical ventilation in sleeping humans. AB - 1. To determine the magnitude and time course of changes in respiratory motor output caused by non-chemical influences, six sleeping subjects underwent assist control mechanical ventilation (ACMV) at increased tidal volume (VT). During ACMV, end-tidal PCO2 (PET,CO2) was either held at normocapnic levels (PET,CO2, 0.6-1.1 mmHg > control) by adding CO2 to the inspirate, or it was allowed to fall (hypocapnia). 2. Each sleeping subject underwent several repeat trials of twenty five ACMV breaths (VT, 1.3 or 2.1 times control; peak flow rate, 30-40 l min-1; inspiratory time, +/- 0.3 s of control). The end-tidal to arterial PCO2 difference throughout normocapnic ACMV at raised VT was unchanged from eupnoeic levels during studies in wakefulness. 3. Normocapnic ACMV at both the smaller and larger increases in VT decreased the amplitude of respiratory motor output, as judged by decreased maximum rate of rise of mask pressure (Pm) (mean dPm/dtmax, 46-68% of control), reduced diaphragmatic EMG (to 55% of control) and reduced VT on the first spontaneous breath after ACMV (to 70% of control). Expiratory time (TE) was slightly prolonged (13-32% > control). This inhibition of amplitude of respiratory motor output progressed over the first five to seven ventilator cycles, was maintained over the remaining 18-20 cycles and persisted for three to five spontaneous breaths immediately following cessation of ACMV. 4. Hypocapnia did not further inhibit respiratory motor output amplitude beyond the effect of normocapnic ACMV at high VT, but did cause highly variable prolongation of TE when PET,CO2 was reduced by greater than 3 mmHg for at least five ventilator cycles. 5. These data in sleeping humans support the existence of a significant, non-chemical inhibitory influence of ACMV at increased VT and positive pressure upon the amplitude of respiratory motor output; this effect is manifested both during and following normocapnic mechanical ventilation. PMID- 10381607 TI - Some properties of dissimilatory nitrate reductases lacking molybdenum and molybdenum cofactor AB - Novel periplasmic and membrane-bound nitrate reductases lacking molybdenum and molybdenum cofactor were isolated from the vanadate-reducing bacterium Pseudomonas isachenkovii, and their properties were studied. Both enzymes have some unusual features, i. e., the individual subunits (130-kD subunit of the membrane-bound enzyme and monomeric 55-kD subunit of the periplasmic enzyme) possess their own nitrate reductase activity. In addition, both enzymes are highly thermostable, their temperature optimum being at 70-80 degrees C, which is unexpectedly high for enzymes from mesophilic bacteria. Similarly to conventional molybdenum-containing nitrate reductases, these isolated enzymes are very sensitive to low concentrations of cyanide and azide. During anaerobic cell growth on medium with nitrate and vanadate, nitrate consumption is followed by a period of vanadate dissimilation, and this period is associated with some structural reorganizations of the nitrate reductases. PMID- 10381608 TI - DNase II in spermatozoa of the loach misgurnus fossilis L AB - A deoxyribonuclease (DNase) which is active at acid pH in the absence of bivalent cations was found in loach spermatozoa. The enzyme was purified by ion-exchange chromatography and partially characterized. The DNase has optimal activity at pH 5.5 and its molecular weight is about 30 kD; its substrate is covalently closed circular duplex DNA, and its product is the corresponding unit-length linear DNA. The DNase is inhibited by MgCl2 and activated by EDTA. Thus, this endoDNase found in loach spermatozoa can be classified as a DNase II. The biological role of this DNase II is discussed. PMID- 10381606 TI - Cutaneous reflexes of the human leg during passive movement. AB - 1. Four experiments tested the hypothesis that movement-induced discharge of somatosensory receptors attenuates cutaneous reflexes in the human lower limb. In the first experiment, cutaneous reflexes were evoked in the isometrically contracting tibialis anterior muscle (TA) by a train of stimuli to the tibial nerve at the ankle. The constancy of stimulus amplitudes was indirectly verified by monitoring M waves elicited in the abductor hallucis muscle. There was a small increase in the reflex excitation (early latency, EL) during passive cycling movement of the leg compared with when the leg was stationary, a result opposite to that hypothesized. There was no significant effect on the magnitude of the subsequent inhibitory reflex component (middle latency, ML), even with increased rate of movement, or on the latency of any of the reflex components. 2. In the second experiment, the two reflex components (EL and ML) elicited in TA at four positions in the movement cycle were compared with corresponding reflexes elicited with the limb stationary at those positions. Despite the markedly different degree of stretch of the leg muscles, movement phase exerted no statistically significant effect on EL or ML reflex magnitudes. 3. In the third experiment, taps to the quadriceps tendon, to elicit muscle spindle discharge, had no effect on the magnitude of ML in TA muscle. The conditioning attenuated EL magnitude for the first 110 ms. Tendon tap to the skin over the tibia revealed similar attenuation of EL. 4. The sural nerve was stimulated at the ankle in the fourth experiment. TA EMG reflex excitatory and inhibitory responses still showed no significant attenuation with passive movement. Initial somatosensory evoked potentials (SEPs), measured from scalp electrodes, were attenuated by movement. 5. The results indicate that there is separate control of transmission in Ia and cutaneous pathways during leg movement. This suggests that modulation of the cutaneous reflex during locomotion is not the result of inhibition arising from motion-related sensory receptor discharge. PMID- 10381609 TI - Nucleotide sequence of the gene and features of the major outer membrane protein of a virulent Rickettsia prowazekii strain. AB - We have determined the nucleotide sequence of the gene for a major outer membrane protein (MOMP) of apparent molecular weight 29.5 kD of the virulent Breinl strain of Rickettsia prowazekii. The gene contains an open reading frame (ORF) that encodes a 282-amino-acid polypeptide with a calculated molecular mass of 31549 daltons. A signal-like peptide sequence is found at the deduced N terminus. A heterologous 29.5-kD antigen expressed in Escherichia coli was shown to be secreted into the periplasm. A database search for similar protein sequences revealed considerable homology of the polypeptide with the E. coli peptidyl prolyl cis/trans isomerase and related proteins of the parvulin family. The genes for MOMP of the virulent Breinl and EVir strains and the vaccine Madrid E strain were amplified using specific primers and cloned into expression vector pQE-30. We found that the polypeptides encoded by the recombinant DNAs do not differ in SDS-PAGE mobility, while the native MOMP of the Breinl strain is known to be different from the corresponding proteins of the Madrid E and EVir strains. Furthermore, no differences within the ORF for the 29.5-kD proteins of the three strains were found by restriction endonuclease analysis of polymerase chain reaction (PCR) products. A possible role of parvulin-like protein (Plp) in the virulence of epidemic typhus agent and the nature of interstrain differences are discussed. Near the plp gene on the opposite strand, an origin of the gene that codes for the SecA subunit of a preprotein translocase was found. PMID- 10381610 TI - Photoelectric responses of oxygen-evolving complexes of photosystem II AB - The generation of a transmembrane electric potential difference induced by a series of laser flashes was studied by the direct electrometrical method in proteoliposomes containing oxygen-evolving particles of photosystem II. In addition to the fast stage of generation of the membrane potential, which is due to electron transfer from the redox active tyrosine residue Tyr-161 (YZ) to the primary quinone acceptor QA, electrogenic stages corresponding to the S1 --> S2 (tau = 30 &mgr;sec), S2 --> S3 (tau = 240 &mgr;sec), and S3 --> S4 --> S0 (tau = 6.2 msec) transitions of the oxygen-evolving complex (OEC) were observed. The amplitudes of the photoelectric responses show that the contribution of the OEC to the overall electrogenicity is small. The parameters of the electrogenic reactions of the OEC as measured in photosystem II preparations containing the peripheral proteins of 23 and 17 kD were similar to those of photosystem II preparations devoid of these peptides. It is concluded that neither the 23- nor the 17-kD proteins are involved in the electrogenic reactions of the OEC. PMID- 10381611 TI - Cellular signalling and free-radical modulating activities of the novel peptidomimetic L-glutamyl-histamine. AB - A novel histamine-containing peptidomimetic, L-glutamyl-histamine (L-Glu-Hist), has been synthesized and characterized as a possible cytokine mimic which might lead to cellular responses of improved specificity. The energy-minimized 3-D conformations of L-Glu-Hist derived from its chemical structure stabilize Fe2+ chelating complexes. L-Glu-Hist concentration-dependently accelerates a decrease in ferrous iron in ferrous sulfate solution and shows ferroxidase-like activity at concentrations less than 3 mM in the phenanthroline assay, whereas in the concentration range 3-20 mM it restricts the availability of Fe2+ to phenanthroline by chelation of iron ions. At low concentrations (less than or about 1 mM), L-Glu-Hist stimulates peroxidation of phosphatidylcholine in liposomes catalyzed by a superoxide anion radical (O2)-generating system (Fe2+ + ascorbate) and, at high concentrations (*10 mM), it suppresses lipid peroxidation (LPO) in liposomes. The stimulation of LPO by L-Glu-Hist is related to its ability at low concentrations (*0.05 mM) to release O2 free radicals as determined by the superoxide dismutase-inhibitable reduction of cytochrome c. The release of O2 by L-Glu-Hist might result from its ferroxidase-like activity, while its inhibition of LPO is due to chelation of Fe2+, prevention of the formation of free radicals, and degradation of lipid hydroperoxides at 5-20 mM L Glu-Hist concentrations. L-Glu-Hist releases O2 at concentrations which stimulate [3H]thymidine incorporation into DNA and proliferation of mouse spleen lymphocytes and also of mononuclear cells from human blood. The induction of lymphocyte proliferation by L-Glu-Hist is dose-dependent in the 0.01-0.05 mM concentration range, although the maximal stimulation of LPO in the O2-dependent system is observed at higher L-Glu-Hist concentrations (*1 mM). Thus, low concentrations of oxygen free radicals released by L-Glu-Hist may provide a very fast, specific, and sensitive trigger for lymphocyte proliferation and immunoregulation. PMID- 10381612 TI - Structure of a neutral O-specific polysaccharide of the bacterium Providencia alcalifaciens O5. AB - A neutral polysaccharide containing D-galactose, 2-acetamido-2-deoxy-D-glucose, and 3-acetamido-3,6-dideoxy-D-glucose (Qui3NAc) in the ratios 2:1:1 was obtained by mild acid degradation of lipopolysaccharide of the bacterium Providencia alcalifaciens O5 followed by gel chromatography and ion-exchange chromatography or treatment with anhydrous hydrogen fluoride. On the basis of full acid hydrolysis, methylation, and 1H- and 13C-NMR spectroscopy, including two dimensional correlation spectroscopy (COSY), total correlation spectroscopy (TOCSY), H-detected heteronuclear 1H,13C single-quantum coherence (HSQC), and nuclear Overhauser effect spectroscopy (NOESY), the following structure of the linear tetrasaccharide repeating unit of the polysaccharide was established: PMID- 10381613 TI - Biochemical predetermination of the NO synthase and nitrite reductase components of the nitric oxide cycle. AB - This review presents some aspects of a concept of cellular evolution bearing a relationship to nitrate--nitrite respiration, the endosymbiosis theory, and the origin of NO synthase and nitrite reductase activity in heme-containing proteins. Analysis of structural and functional unity of the NO synthase and nitrite reductase systems suggests that these systems did not arise without any relation to evolutionarily ancient energetic systems of cells. The use of symmetry principles reveals commonalities among many electron transport chains which in the language of physics is called "invariance". This work also comparatively analyzes the nitric oxide cycle and the known nitrogen cycle. The ideas about evolution of the NO synthase and nitrite reductase systems developed here are clearly compatible with the endosymbiotic theory and the hypothesis that nitrate- nitrite respiration was a precursor of oxygen-dependent respiration. PMID- 10381614 TI - Age-dependent changes of lipid composition in daphnia magna AB - Total lipids, phospholipids, neutral lipids, and fatty acids were studied in three developmental stages of Daphnia magna Straus. Higher amounts of free sterols and all major phospholipids (phosphatidylcholine, phosphatidylethanolamine, and sphingomyelin) were detected in neonates. The content of triacylglycerol stores of juveniles decreased during somatic growth. Detectable amounts of wax esters were found only in adults. The fatty acid unsaturation index was lower in female neonates. The relationship between the lipid contents and the age-related resistance of Daphnia to the environmental factors is discussed. PMID- 10381616 TI - Do protein molecules have a native-like topology in the pre-molten globule state? AB - The process of anion-induced refolding of acid unfolded apomyoglobin with a modified tyrosine residue (Tyr-146(NO2)) has been characterized by intrinsic protein fluorescence. It is shown that under conditions inducing the protein molecule transition into the pre-molten globule state (i.e., at low pH (2.5) and increased sulfate concentration) there is a significant quenching of the tryptophan fluorescence of the protein resulting from efficient energy transfer from the tryptophan residues to the nitrotyrosine residue. This may mean that the protein molecule in the pre-molten globule state has a native-like topology. PMID- 10381615 TI - Opioid agonist modulation of cytoplasmic free Ca2+ level in concanavalin A stimulated mouse lymphocytes. AB - In this study the influence of mu-, delta-, and kappa-selective opioid agonists (DAMGO, DSLET, and dynorphin A (1-13)) on cytoplasmic free Ca2+ ([Ca2+]i) level in normal and concanavalin-A (Con A)-activated mouse lymphocytes was investigated. [Ca2+]i was measured using the fluorescent dye FURA-2AM. The opioid peptides at 10-12-10-7 M induced some increase in [Ca2+]i in non-activated lymphocytes. However, DAMGO and DSLET (10-13-10-7 M) considerably inhibited a Con A-induced increase in [Ca2+]i. The inhibiting effect of both peptides was higher after 20-min preincubation compare to 2-h preincubation. The effect of the kappa agonist dynorphin A (1-13) was significantly different depending on the duration of cell pretreatment and the concentration of the peptide used. After preincubation for 20 min at low concentrations (10-12-10-11 M) it slightly stimulated, while at higher (10-10-10-7 M) concentrations it inhibited lymphocyte response to Con A. After preincubation for 2 h, pronounced stimulation of mitogen induced Ca2+ flux was observed at peptide concentration 10-9 M. The effects of opioids were antagonized by naloxone. These data indicate that functionally active opioid receptors expressed on lymphocytes could be involved in early stages of mitogen activation. PMID- 10381617 TI - Phosphorylation of a low-molecular-weight polypeptide in rat liver mitochondria and dependence of its phosphorylation on mitochondrial functional state. AB - We show that incubation of rat liver mitochondria in the presence of [gamma 32P]ATP results in cAMP-dependent phosphorylation of a low-molecular-weight (3.5 kD) polypeptide (LMWP). This component is tightly bound to the mitochondrial membrane. It is not released into solution after freezing and subsequent thawing of the mitochondrial suspension and does not incorporate 32P from [gamma-32P]ATP in the presence of uncouplers of oxidative phosphorylation. Inhibition of adenine nucleotide transport into the mitochondrial matrix by carboxyatractyloside suppresses phosphorylation of the LMWP. Moderate Ca2+ loading of mitochondria increases both phosphorylation and dephosphorylation of the LMWP. Chelation of Ca2+ by incubation in the presence of EGTA suppresses incorporation of 32P into the LMWP. PMID- 10381618 TI - Investigation of sialic acids and sialyltransferase activity in blood of patients with systemic scleroderma. AB - Systemic scleroderma (SSd) is a connective tissue disorder accompanied by generalized fibrosis. A disturbance of the synthesis and production of matrix glycoproteins, such as collagens, fibronectin, and proteoglycans, by connective tissue cells is typical for this disease. We previously demonstrated a decrease in the ganglioside content of cultured skin fibroblasts from patients with SSd. In this work the contents of sialoglycoproteins and sialoglycolipids in blood sera of patients with SSd were estimated. Simultaneously, the level of asialofetuin-sialyltransferase activity in blood plasma of three groups of patients--those with SSd, Raynaud's phenomenon, and with localized scleroderma- was investigated. CMP-5-acetamido-9-deoxy-9-fluoresceinylthioureidoneuraminic acid was used as a substrate for the enzyme assay. It was shown that the concentration of total sialic acid was increased and the concentration of lipid bound sialic acid was slightly decreased in the blood sera of patients with SSd. A correlation between the lipid-bound sialic acid level and the severity of disease was observed; there was no correlation between severity of disease and total sialic acid. Sialyltransferase assay showed a decrease in the activity level in all three groups of patients. The greatest decrease (2-fold) of this activity was observed in patients with Raynaud's phenomenon. Our data suggest that in SSd and similar diseases the process of glycoconjugate sialylation is disturbed. These changes may considerably affect the mechanisms of regulation of metabolism and cellular interactions. PMID- 10381619 TI - The anion-carrier mediated uncoupling effect of dicarboxylic fatty acids in liver mitochondria depends on the position of the second carboxyl group. AB - Effects of dicarboxylic fatty acids with varying positions of the carboxyl groups on respiration and membrane potential of liver mitochondria were studied. Tetradecylmalonic acid (a fatty acid with two carboxyl groups in the alpha position) efficiently uncoupled oxidative phosphorylation similarly to palmitic acid with the same number of carbon atoms. Similarly to the uncoupling by palmitic acid, the coupling effects of carboxyatractylate and glutamate changed reciprocally with changes in pH of the incubation medium: on increasing the pH from 7.0 to 7.8, the coupling effect of carboxyatractylate increased and that of glutamate decreased. A dicarboxylic fatty acid with the second carboxyl at the end of the alkyl chain in the omega-position (alpha, omega-tetradecyldicarboxylic acid) stimulated respiration of the mitochondria at a significantly higher concentration than myristic acid with the same number of carbon atoms, but unlike the latter nearly failed to decrease the transmembrane potential DeltaPsi. Neither carboxyatractylate nor glutamate inhibited the respiration stimulated by this dicarboxylic fatty acid. PMID- 10381620 TI - Structural organization of membrane and soluble forms of somatic angiotensin converting enzyme. AB - The catalytic activity and quaternary structure of soluble (s) and membrane (m) forms of angiotensin-converting enzyme (ACE) were studied in reversed micelles of ternary system Aerosol OT--water--octane. The profile of the dependence of the catalytic activity of the two enzyme forms on the degree of surfactant hydration (micellar size) had several optima corresponding to the function of various active oligomeric enzyme forms; the curves for the s- and m-forms of ACE were different. Data of sedimentation analysis prove that in reversed micelles, s-ACE can exist as monomers, dimers, or tetramers depending on the hydration degree, and the m-form is present as dimers and tetramers only. The values of the kinetic parameters for the hydrolysis of the substrate furylacryloyl-Phe-Gly-Gly by all the enzyme forms were determined, and the data indicate that the activity of the m-form is enhanced by oligomerization. The ACE activity strongly depends on the medium; it is higher when ACE is in contact with matrix or other enzyme molecules. PMID- 10381621 TI - A site-specific endonuclease from the thermophilic strain Bacillus species F4. AB - A strain producing the site-specific endonuclease BspF4I was found during screening of thermophilic bacteria isolated from soil. The restriction endonuclease, free from contaminant nonspecific nucleases, was purified using three steps of column chromatography--on hydroxyapatite, blue agarose, and DEAE Trisacryl. The enzyme is stable on storage and exhibits maximal activity at 48-56 degrees C in the presence of albumin in buffer containing 10 mM Tris-HCl (pH 7.5) and 10 mM MgCl2. BspF4I recognizes the sequence 5;-GGNCC-3; on DNA and is an isomer and not an isoschizomer of the endonuclease Sau96I. Unlike the prototype, BspF4I does not cleave the site in a defined way. A strand with purine in the center of the sequence is cleaved after the first G, as in the case of the prototype, while the strand with pyrimidine is cleaved either before or after the first G. PMID- 10381622 TI - Phosphorescence of intermediates of the terminal stage of chlorophyll biosynthesis in plants AB - Phosphorescence (radiative triplet state deactivation) of the intermediates involved in chlorophyll a photobiosynthesis from protochlorophyllide (Pchld) was found in greening leaves at 77K. Photoactive Pchld forms showed phosphorescence with the major spectral bands at 920 and 970 nm and 2.5-3 msec lifetime. Photochemical formation of the primary non-fluorescent intermediates (NFI) was accompanied by quenching of Pchld fluorescence and phosphorescence, phosphorescence quenching being less efficient by a factor of two. The primary chlorophyllide form Chld 684/676 that appeared as a result of NFI dark transformation showed phosphorescence with a maximum at 980 nm and 2 msec lifetime. Secondary chlorophyllide forms Chld 690/680 and Chld 695/685, the products of photochemical Chld 684/676 transformation and intermediates in biosynthesis of chlorophyll of the light harvesting complex, emitted phosphorescence with maxima at 990-995 and 1005-1010 nm and 1-2 msec lifetimes. Chlorophyll Chl 675/668, the major product of the non-photochemical "side" reaction of Chld 684/676 that probably underlies the biogenesis of photosystem II, exhibited phosphorescence with a maximum at 954-960 nm and 2 msec lifetime. Analysis of the difference spectra of the side reaction products revealed a minor 930 nm phosphorescence band that might correspond to pheophytin a formed via the "side" reaction. Triplet states of the intermediates were not quenched by carotenoids, and the quantum yields of their population at 77K were not less than 10%. Under physiological conditions the triplet intermediates might be involved in photochemical reactions, in particular, in singlet oxygen photogeneration and photodestruction of the photosynthetic apparatus. PMID- 10381623 TI - A novel adenovirus E1B19K-binding protein B5 inhibits apoptosis induced by Nip3 by forming a heterodimer through the C-terminal hydrophobic region. AB - The adenovirus E1B19K protein inhibits apoptosis induced by E1A and other divergent signals. The cellular proteins that interact with E1B19K have been analyzed by isolating cDNA clones by the yeast two hybrid system. One of these clones encodes B5 which consists of 219 amino acid residues and contains the putative BH3 and transmembrane regions. B5 binds strongly to Nip3 and itself, weakly to E1B19K, but not to Bcl-2 and localizes in nuclear envelope, endoplasmic reticulum and mitochondria. B5 has sequence homology with Nip3 in the middle and C-terminal regions, but not in the N-terminal region. Unlike other E1B19K binding BH3 proteins so far characterized, B5 does not induce apoptosis, but inhibits apoptosis induced by Nip3. However the deletion mutant B5Delta1-31 lacking the N terminus does induce apoptosis, although weaker than does Nip3, suggesting that the N-terminal region is masking the apoptosis-inducing capacity of B5. PMID- 10381624 TI - Purification and catalytic properties of human caspase family members. AB - Members of the caspase family of cysteine proteases are known to be key mediators of mammalian inflammation and apoptosis. To better understand the catalytic properties of these enzymes, and to facilitate the identification of selective inhibitors, we have systematically purified and biochemically characterized ten homologues of human origin (caspases 1 - 10). The method used for production of most of these enzymes involves folding of active enzymes from their constituent subunits which are expressed separately in E. coli, followed by ion exchange chromatography. In cases where it was not possible to use this method (caspase-6 and -10), the enzymes were instead expressed as soluble proteins in E. coli, and partially purified by ion exchange chromatography. Based on the optimal tetrapeptide recognition motif for each enzyme, substrates with the general structure Ac-XEXD-AMC were used to develop continuous fluorometric assays. In some cases, enzymes with virtually identical tetrapeptide specificities have kcat/Km values for fluorogenic substrates that differ by more than 1000-fold. Using these assays, we have investigated the effects of a variety of environmental factors (e.g. pH, NaCl, Ca2+) on the activities of these enzymes. Some of these variables have a profound effect on the rate of catalysis, a finding that may have important biological implications. PMID- 10381625 TI - A novel adenoviral vector expressing human Fas/CD95/APO-1 enhances p53-mediated apoptosis. AB - Recent evidence suggests an intriguing link between p53 and the Fas pathway. To evaluate this association further, we utilized a recombinant adenoviral vector (AdWTp53) to overexpress wild-type p53 in lung cancer (A549, H23, EKVX and HOP92) and breast cancer (MDA-MB-231 and MCF-7) cell lines and observed an increase in the Fas/CD95/APO-1 protein levels. Furthermore, this increase correlated with the sensitivity of the cell lines to p53-mediated cytotoxicity. To examine the effects of Fas over-expression in cells resistant to p53 over-expression, we constructed AdFas, an adenoviral vector capable of transferring functional human Fas to cancer cells. Interestingly, infection of p53-resistant MCF-7 cells with AdFas sensitized them to p53-mediated apoptosis. These studies indicate that combined over-expression of Fas and wild-type p53 may be an effective cancer gene therapy approach, especially in cells relatively resistant to p53 over expression. PMID- 10381627 TI - Nucleolar segregation during apoptosis of haemopoietic stem cell line FDCP-Mix. AB - The programmed elimination of cells during apoptosis is distinct from necrosis both morphologically and biochemically. Currently, the morphological description of apoptosis discriminates between the segregation of the nucleolus and the so called 'chromatin condensation'. The latter originates from observations of electron dense material adjacent to the nuclear envelope of apoptotic nuclei. Although there is ample evidence for an involvement of DNA in electron dense marginations, their true nature is still unknown. By studying apoptosis in FDCP Mix, a pluripotent murine haemopoietic stem cell line, we found morphological and histochemical evidence that electron dense material at the nuclear envelope (NE) has emerged as a result of the segregation of nucleoli in association with the nuclear membrane. The remaining electron dense and homogenous bulk of the nucleolus labels for RNAse-gold, but even more intensely for DNAse-gold, and therefore could possibly be mistaken as 'condensed chromatin' in the light microscope. The labelling of the electron dense material for DNase-gold in FDCP Mix could be explained by a migration of DNA into the bulk of the nucleoli at an early stage of cell death. PMID- 10381626 TI - Selective inhibition of apoptosis by TPA-induced differentiation of U937 leukemic cells. AB - U937 leukemic cells treated for 24 h with 16 nM 12-O-tetradecanoylphorbol 13 acetate (TPA), that induces their macrophagic terminal differentiation, become resistant to etoposide-induced apoptosis. Exposure of undifferentiated U937 cells to 50 microM etoposide for 6 h, that triggers apoptosis in 80% cells, activates procaspase-2L, -3 and -8, induces the mitochondrial release of cytochrome c and decreases Mcl-1 expression without modifying Bcl-2, Bcl-xL and Bax protein levels. All these events are inhibited in TPA-differentiated U937 cells that are also resistant to vinblastine-induced and Fas-mediated cell death. Interestingly, these cells are not inherently resistant to apoptosis induction. Exposure of TPA differentiated U937 cells to 0.8 microg/ml cycloheximide for 24 h, that triggers apoptosis in 50% cells, activates procaspase-2L, -3 and -8, induces the mitochondrial release of cytochrome c and decreases Bcl-xL expression without modifying Bcl-2, Mcl-1 and Bax protein levels. All these events are not observed in undifferentiated cells treated in similar conditions. These results indicate that the apoptotic pathway that involves the release of cytochrome c from mitochondria and the cleavage of procaspases remains functional in TPA differentiated cells. PMID- 10381628 TI - N-acetylcysteine blocks apoptosis induced by N-alpha-tosyl-L-phenylalanine chloromethyl ketone in transformed T-cells. AB - The serine protease inhibitor N-alpha-tosyl-L-phenylalanine chloromethyl ketone (TPCK) can interfere with cell-cycle progression and has also been shown either to protect cells from apoptosis or to induce apoptosis. We tested the effect of TPCK on two transformed T-cell lines. Both Jurkat T-cells and Theileria parva transformed T-cells were shown to be highly sensitive to TPCK-induced growth arrest and apoptosis. Surprisingly, we found that the thiol antioxidant, N acetylcysteine (NAC), as well as L- or D-cysteine blocked TPCK-induced growth arrest and apoptosis. TPCK inhibited constitutive NF-kappaB activation in T. parva-transformed T-cells, with phosphorylation of IkappaBalpha and IkappaBbeta being inhibited with different kinetics. TPCK-mediated inhibition of IkappaB phosphorylation, NF-kappaB DNA binding and transcriptional activity were also prevented by NAC or cysteine. Our observations indicate that apoptosis and NF kappaB inhibition induced by TPCK result from modifications of sulphydryl groups on proteins involved in regulating cell survival and the NF-kappaB activation pathway(s). PMID- 10381629 TI - Synthesis of procaspases-3 and -7 during apoptosis in prostate cancer cells. AB - Cells differ in the time required to execute cell death after receipt of a death signal. One reason may be the requirement for de novo synthesis of components of the death pathway. TSU-Pr1 prostate cancer cells treated with okadaic acid demonstrated activation of caspase-3, PARP cleavage, and nuclear fragmentation by 24 h and apoptosis by 72 h. Levels of procaspase-3 and procaspase-7, the precursor molecules of two effector caspases, were not depleted during apoptosis. Levels of procaspase-3 and -7 mRNA increased steadily in TSU-Pr1 cells up to 72 h after exposure to okadaic acid. Nuclear run-off experiments showed that the increase in mRNA was not due to transcriptional activation of caspase-3 and -7 mRNA. Antisense caspase-3 and caspase-7 oligodeoxynucleotides caused a depletion of procaspases-3 and -7 and a delay in apoptosis of TSU-Pr1 cells. Caspase antisense oligodeoxynucleotides inhibited apoptosis to a similar extent as peptide inhibitors of cysteine proteases. Synthesis of procaspases-3 and -7 was necessary to sustain programmed cell death in TSU-Pr1 prostate cancer cells. PMID- 10381630 TI - Expression and biological activity of X-linked inhibitor of apoptosis (XIAP) in human malignant glioma. AB - The inhibitor-of-apoptosis (IAP) proteins are a novel family of antiapoptotic proteins that are thought to inhibit cell death via direct inhibition of caspases. Here, we report that human malignant glioma cell lines express XIAP, HIAP-1 and HIAP-2 mRNA and proteins. NAIP was not expressed. IAP proteins were not cleaved during CD95 ligand (CD95L)-induced apoptosis, and loss of IAP protein expression was not responsible for the potentiation of CD95L-induced apoptosis when protein synthesis was inhibited. LN-18 cells are highly sensitive to CD95 mediated apoptosis, whereas LN-229 cells require co-exposure to CD95L and a protein synthesis inhibitor, CHX, to acquire sensitivity to apoptosis. Adenoviral XIAP gene transfer blocked caspase 8 and 3 processing in both cell lines in the absence of CHX. Apoptosis was blocked in the absence and in the presence of CHX. However, XIAP failed to block caspase 8 processing in LN-229 cells in the presence of CHX. There was considerable overlap of the effects of XIAP on caspase processing with those of BCL-2 and the viral caspase inhibitor crm-A. These data define complex regulatory mechanisms for CD95-mediated apoptosis in glioma cells and indicate that there may be a distinct pathway of death receptor-mediated apoptosis that is readily activated when protein synthesis is inhibited. The constitutive expression of natural caspase inhibitors may play a role in the resistance of these cells to apoptotic stimuli that directly target caspases, including radiochemotherapy and immune-mediated tumor cell lysis. PMID- 10381631 TI - The fate of E- and P-cadherin during the early stages of apoptosis. AB - Caspases are responsible for the proteolysis of many cytoskeletal proteins in apoptotic cells. It has been demonstrated here that during cisplatin-induced apoptosis of human embryo retinoblasts both E- and P-cadherin were degraded by caspases, giving initially major polypeptide products of apparent molecular weights 48 K and 104 K respectively. This proteolysis occurred over a similar time-scale to the observed degradation of PARP and to the onset of DNA fragmentation but appreciably later than p53 induction and cleavage of Mdm2 and p21. Addition of caspase inhibitors such as Z-VAD-FMK inhibited apoptosis and cadherin degradation. Co-immunoprecipitation studies carried out on viable cells confirmed previously observed complexes between cadherins and alpha and beta catenin and between the catenins themselves. These interactions were sustained in apoptotic cells as long as the protein components remained intact. Using confocal microscopy it has been shown that cytoskeletal changes associated with apoptosis precede degradation of catenins and cadherins by several hours. In particular, after addition of cisplatin relatively rapid (within 3 h) re-localization of adherens junction proteins from the cell periphery to the cytoplasm was observed whereas little cadherin or catenin degradation occurred until 10 h. We conclude that neither caspase-mediated degradation of cytoskeletal components nor disruption of adherens junction protein-protein interactions is required for morphological change. PMID- 10381632 TI - The role of the ubiquitin-proteasome pathway in apoptosis. AB - Coordinated intracellular protein degradation mediated by the ubiquitin proteasome pathway is crucial to a vast array of cellular processes including orderly progression through the mitotic cycle. Similarly important to both the fates of individual cells, as well as to the normal function of multicellular organisms, is the process of apoptosis, or programmed cell death. Execution of this latter process has been known for some time to be intimately associated with the activity of caspases, a family of proteases related to interleukin-1-beta converting enzyme. Evidence is now accumulating, however, that the ubiquitin proteasome system itself plays an important role in apoptosis, and some of the cellular pathways that are impacted upon by the proteasome, and may lead to apoptosis, are beginning to be dissected. This review provides a summary of the experimental basis by which components of the ubiquitin-proteasome pathway have been linked to apoptosis, and attempts are made to formulate a hypothesis about its role in this process. PMID- 10381633 TI - bFGF inhibits the activation of caspase-3 and apoptosis of P19 embryonal carcinoma cells during neuronal differentiation. AB - P19 embryonal carcinoma (EC) cells undergo apoptosis during neuronal differentiation induced by all-trans retinoic acid (RA). Caspase-3-like proteases are activated and involved in the apoptosis of P19 EC cells during neuronal differentiation.1 Recently it has been shown that growth factor signals protect against apoptosis by phosphorylation of Bad. Phosphorylated Bad, an apoptotic member of the Bcl-2 family, cannot bind to Bcl-xL and results in Bcl-xL homodimer formation and subsequent antiapoptotic activity. In the present study, we demonstrate that this system is used generally to protect against apoptosis during neuronal differentiation. Bcl-xL inhibited the activation of caspase-3 like proteases. Basic fibroblast growth factor (bFGF) inhibited more than 90% of the caspase-3-like activity, inhibited processing of caspase-3 into its active form, and inhibited DNA fragmentation. bFGF activated phosphatidyl-inositol-3 kinase (PI3K) and stimulated the phosphorylation of Bad. Phosphorylation was inhibited by wortmannin, an inhibitor of PI3K and its downstream target Akt. Thus, Bad is a target of the FGF receptor-mediated signals involved in the protection against activation of caspase-3. PMID- 10381634 TI - Etoposide-induced activation of c-jun N-terminal kinase (JNK) correlates with drug-induced apoptosis in salivary gland acinar cells. AB - We have examined the ability of etoposide to induce apoptosis in two recently established rat salivary acinar cell lines. Etoposide induced apoptosis in the parotid C5 cell line as evidenced by the appearance of cytoplasmic blebbing and nuclear condensation, DNA fragmentation and cleavage of PARP. Etoposide also induced activation of c-jun N-terminal kinase (JNK) in parotid C5 cells by 4 h after treatment, with maximal activation at 8 - 10 h. Coincident with activation of JNK, the amount of activated ERK1 and ERK2 decreased in etoposide-treated parotid C5 cells. In contrast to the parotid C5 cells, the vast majority of submandibular C6 cells appeared to be resistant to etoposide-induced apoptosis. Likewise, activation of JNKs was not observed in etoposide-treated submandibular C6 cells, and the amount of activated ERK1 and ERK2 decreased only slightly. Etoposide treatment of either cell line had no effect upon the activation of p38. Treatment of the parotid C5 cells with Z-VAD-FMK, a caspase inhibitor, inhibited etoposide-induced activation of JNK and DNA fragmentation. These data suggest that etoposide may induce apoptosis in parotid C5 cells by activating JNKs and suppressing the activation of ERKs, thus creating an imbalance in these two signaling pathways. PMID- 10381635 TI - Manganese induces apoptosis of human B cells: caspase-dependent cell death blocked by bcl-2. AB - Manganese ions block apoptosis of phagocytes induced by various agents. The prevention of apoptosis was attributed to the activation of manganous superoxide dismutase (Mn-SOD) and to the antioxidant function of free Mn2+ cations. However, the effect of Mn2+ on B cell apoptosis is not documented. In this study, we investigated the effects of Mn2+ on the apoptotic process in human B cells. We observed that Mn2+ but not Mg2+ or Ca2+, inhibited cell growth and induced apoptosis of activated tonsilar B cells, Epstein Barr virus (EBV)-negative Burkitt's lymphoma cell lines (BL-CL) and EBV-transformed B cell lines (EBV-BCL). In the same conditions, no apoptosis was observed in U937, a monoblastic cell line. Induction of B cell apoptosis by Mn2+ was time- and dose-dependent. The cell permeable tripeptide inhibitor of ICE family cysteine proteases, zVAD-fmk, suppressed Mn2+-induced apoptosis. Furthermore, Mn2+ triggered the activation of interleukin-1beta converting enzyme (ICE/caspase 1), followed by the activation of CPP32/Yama/Apopain/caspase-3. In addition, poly-(ADP-ribose) polymerase (PARP), a cellular substrate for CPP32 protease was degraded to generate apoptotic fragments in Mn2+-treated B cell lines. The inhibitor, zVAD-fmk suppressed Mn2+-triggered CPP32 activation and PARP cleavage and apoptosis. These results indicate that the activation of caspase family proteases is required for the apoptotic process induced by Mn2+ treatment of B cells. While the caspase-1 inhibitor YVAD was unable to block apoptosis, the caspase-3 specific inhibitor DEVD-cmk, partially inhibited Mn2+-induced CPP32 activation, PARP cleavage and apoptosis of cells. Moreover, Bcl-2 overexpression in BL-CL effectively protected cells from apoptosis and cell death induced by manganese. This is the first report showing the involvement of Mn2+ in the regulation of B lymphocyte death presumably via a caspase-dependent process with a death-protective effect of Bcl 2. PMID- 10381636 TI - Dynamic correlation of apoptosis and immune activation during treatment of HIV infection. AB - T cells from HIV infected patients undergo spontaneous apoptosis at a faster rate than those from uninfected patients, are abnormally susceptible to activation induced cell death (AICD), and undergo increased apoptosis in response to Fas receptor ligation. These observations have led to the hypothesis CD4 T cell apoptosis may be a mechanism of CD4 T cell depletion and the pathogenesis of AIDS. Successful treatment of HIV infected patients is accompanied by quantitative and qualitative improvements in immune function reflecting at least partial reversibility of the underlying pathogenesis of HIV. In this report we correlate improvements in markers of immune function with a decrease in apoptosis, and changes in its regulation. Therapy with nelfinavir plus saquinavir in combination with two nucleoside analogue inhibitors of reverse transcriptase dramatically reduces plasma viremia and increases CD4 T cell counts. Coincident with these improvements, CD38 and HLA-DR coexpression on both CD4 and CD8 T cells decrease, and CD45RA and CD62L coexpression increase. Furthermore, spontaneous apoptosis decreases in both CD4 and CD8 T cells (CD4 apoptosis 17.4 vs 2.6%, P=0.005; CD8 apoptosis 15.0 vs 1.0%, P<0.001), as does both Fas mediated apoptosis (CD4 apoptosis 19.0 vs 3.5%, P=0.03; CD8 apoptosis 13.7 vs 1.5%, P=0.002) and CD3 induced AICD (CD4 apoptosis 13.7 vs 3.2%, P=0.001; CD8 apoptosis 29 vs 2.2%, P=0.08). Changes in apoptosis are not associated with changes in Fas receptor expression, but are significantly correlated with changes in activation marker profiles. Although this suggests a possible regulatory role for the apoptosis inhibitory protein FLIP, direct assessment did not reveal quantitative differences in FLIP expression between apoptosis resistant PBL's from HIV negative patients, and apoptosis sensitive PBL's from HIV positive patients. These findings support the hypothesis that apoptosis mediates HIV induced CD4 T cell depletion, but indicate the need for further studies into the molecular regulation of HIV induced apoptosis. PMID- 10381637 TI - Induction of apoptosis by all-trans retinoic acid in the human myeloma cell line RPMI 8226 and negative regulation of some of its typical morphological features by dexamethasone. AB - We investigated the effects of all-trans retinoic acid (RA) and dexamethasone (Dex) on the in vitro growth of the human myeloma cell line RPMI 8226. RA inhibited RPMI 8226 cell growth by both antiproliferative effect and induction of apoptosis. Typical morphological and biochemical characteristics of apoptosis including chromatin condensation, apoptotic bodies formation and internucleosomal DNA cleavage were detected after 4 days of treatment with 1 microM RA. In situ TUNEL assay demonstrated that DNA cleavage preceded chromatin condensation. The expression of tissue transglutaminase (tTG), an enzyme proposed to play a role in apoptosis was induced with RA, as shown by both enzymatic assay and in situ immunofluorescence detection. Dex, when used alone, had no effect on cell growth and apoptosis. When combined to RA, Dex did not interfere with the RA-dependent inhibition of cell proliferation, but unexpectedly inhibited both quantitatively and qualitatively several morphological and biochemical features of the apoptosis induced by RA. Dex did not affect RA-induced DNA breaks formation but impeded the progression of chromatin condensation and the formation of apoptotic bodies. Interestingly, Dex also inhibited the RA-dependent induction of tTG. RU486, a glucocorticoid antagonist, counteracted all Dex effects. Taken together these data demonstrate that key cytoplasmic and nuclear events occurring during apoptosis are differentially regulated by RA and Dex in myeloma cell line RPMI 8226. PMID- 10381638 TI - Death pathway genes Fas (Apo-1/CD95) and Bik (Nbk) show no mutations in colorectal carcinomas. PMID- 10381639 TI - Induction of CD95 ligand and apoptosis by doxorubicin is modulated by the redox state in chemosensitive- and drug-resistant tumor cells. AB - Induction of CD95 ligand (CD95-L) may contribute to drug-induced apoptosis in chemosensitive leukemias and solid tumors. Here we report that induction of CD95 L and apoptosis by doxorubicin in leukemic and neuroblastoma cells is regulated by the redox state and reactive oxygen species (ROS). Preincubation of chemosensitive cells with antioxidants such as N-acetyl-cysteine (NAC) or glutathione (GSH), significantly reduced doxorubicin-induced apoptosis, hyperexpression of ROS, loss of mitochondrial membrane potential (DeltaPsim) and upregulation of CD95-L expression. Doxorubicin-resistant cells exhibited higher levels of GSH in comparison to chemosensitive cells and were deficient in hyperproduction of ROS, loss of DeltaPsim and upregulation of CD95-L in response to cytotoxic drugs. Downregulation of intracellular GSH concentrations reversed deficient drug-induced hyperproduction of ROS and CD95-L upregulation. In addition, overexpression of Bcl-XL in CEM cells blocked doxorubicin-triggered ROS and CD95-L expression. These findings suggest that induction of CD95-L by cytotoxic drugs is modulated by the cellular redox state and mitochondria derived ROS. PMID- 10381640 TI - Caspases in T-cell receptor-induced thymocyte apoptosis. AB - Apoptosis eliminates inappropriate or autoreactive T lymphocytes during thymic development. Intracellular mediators involved in T-cell receptor (TCR)-mediated apoptosis in developing thymocytes during negative selection are therefore of great interest. Caspases, cysteine proteases that mediate mature T-cell apoptosis, have been implicated in thymocyte cell death, but their regulation is not understood. We examined caspase activities in distinct thymocyte subpopulations that represent different stages of T-cell development. We found caspase activity in CD4+CD8+ double positive (DP) thymocytes, where selection involving apoptosis occurs. Earlier and later thymocyte stages exhibited no caspase activity. Only certain caspases, such as caspase-3 and caspase-8-like proteases, but not caspase-1, are active in DP thymocytes in vivo and can be activated when DP thymocytes are induced to undergo apoptosis in vitro by TCR crosslinking. Thus, specific caspases appear to be developmentally regulated in thymocytes. PMID- 10381641 TI - Fas mediated apoptosis of human Jurkat T-cells: intracellular events and potentiation by redox-active alpha-lipoic acid. AB - Activation of caspases is required in Fas receptor mediated apoptosis. Maintenance of a reducing environment inside the cell has been suggested to be necessary for caspase activity during apoptosis. We explored the possibility to potentiate Fas mediated killing of tumor cells by alpha-lipoic acid (LA), a redox active drug and nutrient that is intracellularly reduced to a potent reductant dihydrolipoic acid. Treatment of cells with 100 microM LA for 72 h markedly potentiated Fas-mediated apoptosis of leukemic Jurkat cells but not that of peripheral blood lymphocytes from healthy humans. In Jurkat, Fas activation was followed by rapid loss of cell thiols, decreased mitochondrial membrane potential, increased [Ca2+]i and increased PKC activity; all these responses were potentiated in LA pretreated cells. PKCdelta played an important role in mediating the effect of LA on Fas-mediated cell death. In response to Fas activation LA treatment potentiated caspase 3 activation by over 100%. The ability of LA to potentiate Fas mediated killing of leukemic cells was abrogated by a caspase 3 inhibitor suggesting that increased caspase 3 activity in LA treated Fas-activated cells played an important role in potentiating cell death. This work provides first evidence showing that inducible caspase 3 activity may be pharmacologically up-regulated by reducing agents such as dihydrolipoic acid. PMID- 10381642 TI - Cleavage and nuclear translocation of the caspase 3 substrate Rho GDP dissociation inhibitor, D4-GDI, during apoptosis. AB - While investigating endonucleases potentially involved in apoptosis, an antisera was raised to bovine deoxyribonuclease II, but it recognized a smaller protein of 26 kDa protein in a variety of cell lines. The 26 kDa protein underwent proteolytic cleavage to 22 kDa concomitantly with DNA digestion in cells induced to undergo apoptosis. Sequencing of the 26 kDa protein identified it as the Rho GDP-dissociation inhibitor D4-GDI. Zinc, okadaic acid, calyculin A, cantharidin, and the caspase inhibitor z-VAD-fmk, all prevented the cleavage of D4-GDI, DNA digestion, and apoptosis. The 26 kDa protein resided in the cytoplasm of undamaged cells, whereas following cleavage, the 22 kDa form translocated to the nucleus. Human D4-GDI, and D4-GDI mutated at the caspase 1 or caspase 3 sites, were expressed in Chinese hamster ovary cells which show no detectable endogenous D4-GDI. Mutation at the caspase 3 site prevented D4-GDI cleavage but did not inhibit apoptosis induced by staurosporine. The cleavage of D4-GDI could lead to activation of Jun N-terminal kinase which has been implicated as an upstream regulator of apoptosis in some systems. However, the results show that the cleavage of D4-GDI and translocation to the nucleus do not impact on the demise of the cell. PMID- 10381643 TI - Apoptosis at the time of embryo implantation in mouse and rat. AB - The aim of this review is to summarize the information currently available regarding the occurrence of apoptosis in the developing embryo and in the receptive uterus during the peri-implantation period of gestation. Cell death is detected in the inner cell mass of late pre-implantation embryos as the result of an eliminative process that helps trim the embryonic cell lineages of surplus or dysfunctional stem cells. Cell death is also detected in the epiblastic core of early post-implantation embryos, where the process is implicated in the formation of the pro-amniotic cavity. On the maternal side, uterine epithelial cells situated around the attachment site undergo cell death during the initial phase of implantation in order to facilitate embryo anchorage and access to maternal blood supply. Uterine stromal cells closest to the implantation chamber first transform into decidual cells and then commit suicide to make room for the rapidly growing embryo. Although apoptosis is well recognized as a crucial determinant of successful peri-implantation development, our understanding of the cellular and molecular mechanisms regulating this process clearly lags behind the comprehension of cell death control in other systems. PMID- 10381644 TI - Truncated products of the vestigial proliferation gene induce apoptosis. AB - The vestigial (vg) gene in D. melanogaster, whose mutant phenotype is characterized by wing atrophy, encodes a novel nuclear protein involved in cell proliferation. The original vg mutant (vgBG) displays massive apoptosis in the wing imaginal disc. Here we tested the hypothesis that the vg mutant phenotype could be due: (i) to lack of cell proliferation in null mutants due to the absence of the Vg product and, (ii) to apoptosis in vgBG and other mutants due to the presence of a major Vg truncated product. In agreement with our hypothesis no cell death was observed in null vg mutants, and the anticell death baculovirus P35 product is unable to rescue the mutant phenotype caused by absence of the Vg product. In addition, expression of the antiproliferative gene dacapo, the homolog of p21, induces a mutant wing phenotype without inducing cell death. In contrast the wing phenotype of the original vg mutant could be reproduced by the ectopic expression of the reaper cell death gene when expressed by vg regulatory sequences. In agreement with the hypothesis, the classic vg mutant spontaneously displays an increase in reaper expression in the wing disc and its phenotype can be partially rescued by the P35 product. Finally, we showed that ectopic expression of a truncated Vg product is able on its own to induce ectopic cell death and reaper expression. Our results shed new light on the function of the vg gene, in particular, they suggest that the normal and truncated products affect vg target genes in different ways. PMID- 10381645 TI - ATP and adenosine prevent via different pathways the activation of caspases in apoptotic AKR-2B fibroblasts. AB - Confluent AKR-2B fibroblasts rapidly disintegrate after serum deprivation.27 ATP or adenosine added immediately after serum removal afforded substantial protection against cell death even for a long period of 24 h. ED50 values were 14 and 110 microM for ATP and adenosine, respectively. In the presence of 5 microg/ml cycloheximide the protective effect of both substances was suppressed, indicating that protein synthesis is required. The protective effect of ATP was highly specific since among numerous tested derivatives only ATP-[gamma-S] exhibited a substantial protective effect. The ability of ATP and adenosine to modulate cell division was analyzed. Both substances did not exhibit any mitogenic effect. Adenosine completely blocked PDGF-BB induced cell division, whereas ATP had no effect. Unlike adenosine, ATP strongly stimulated Ca2+-release from intracellular stores. On the other hand, adenosine stimulated an increase in the intracellular concentration of cAMP from 0.4 - 1.5 microM, whereas ATP decreased the content below 0.1 microM. ATP stimulated the phosphorylation of MAP kinase, RSK and p70S6-kinase; adenosine was inactive. After complexation of [Ca2+]i the protective effect of ATP was greatly lost while adenosine was still active. Surprisingly neither ATP nor adenosine caused an activation of PKC isoforms. After incubation with pertussis toxin, the protection by ATP was reduced indicating an involvement of Gi-proteins in the signal transduction induced by ATP. Our results indicate that ATP as well as adenosine are potent inhibitors of cell death caused by serum deprivation and that this protective effect apparently occurs via distinct pathways. However, both pathways must converge at the point of caspase activation, since the stimulation of DEVDase- and VEIDase-activities, respectively, are suppressed by either ATP or adenosine. PMID- 10381646 TI - Survival activity of Bcl-2 homologs Bcl-w and A1 only partially correlates with their ability to bind pro-apoptotic family members. AB - Certain Bcl-2 family members promote cell survival, whereas others promote apoptosis. To explore further how heterodimerization of opposing members affects survival activity, we have compared the abilities of the anti-apoptotic Bcl-w and A1 to bind to the pro-apoptotic Bax, Bak, Bad and Bik and to protect cells from their cytotoxic action. Bcl-w co-immunoprecipitated from cell lysates with Bax, Bak, Bad and Bik, but A1 bound only Bak and Bik. Mutation of A1 at a highly conserved glycine within the BH1 domain prevented binding, but the comparable Bcl w mutant still bound Bak, Bad and Bik, indicating that the glycine is not essential for all heterodimerization. Bcl-w and A1 protected against apoptosis induced by over-expression of Bax or Bad but not that induced by Bak or Bik. With several gene pairs, binding and protection were discordant. The results may reflect critical threshold affinities but also suggest that certain pro-apoptotic proteins may also contribute to apoptosis by a mechanism independent of binding pro-survival proteins. PMID- 10381647 TI - Apoptosis: cell death defined by caspase activation. PMID- 10381648 TI - Additional complexity in p73: induction by mitogens in lymphoid cells and identification of two new splicing variants epsilon and zeta. PMID- 10381649 TI - Different p73 splicing variants are expressed in distinct tumour areas of a multifocal neuroblastoma. PMID- 10381650 TI - Caspases and apoptosis - biology and terminology. PMID- 10381651 TI - Role for cyclin D1 in UVC-induced and p53-mediated apoptosis. AB - DNA damaging agents such as ultraviolet (UV) induce cell cycle arrest followed by apoptosis in cells where irreparable damage has occurred. Here we show that during early phase G1 arrest which occurs in UV-irradiated human U343 glioblastoma cells, there are (1) decreases in cyclin D1 and cdk4 levels which parallel a loss of S-phase promoting cyclin D1/cdk4 complexes, and (2) increases in p53 and p21 protein levels. We also show that the late phase UV-induced apoptosis of U343 cells occurs after cell cycle re-entry and parallels the reappearance of cyclin D1 and cdk4 and cyclin D1/cdk4 complexes. These findings suggest that cyclin D1 can abrogate UV-induced G1 arrest and that the p53 mediated apoptosis that occurs in these cells is dependent on cyclin D1 levels. We examined these possibilities using U343 cells that ectopically express cyclin D1 and found that indeed cyclin D1 can overcome the cell cycle arrest caused by UV. Moreover, the appearance of p53 protein and the induction of apoptosis in UV irradiated cells was found to be dependent on the level of ectopically expressed cyclin D1. These findings, therefore, indicate that expression of cyclin D1 following DNA damage is essential for cell cycle re-entry and p53-mediated apoptosis. PMID- 10381652 TI - Apoptosis without caspases: an inefficient molecular guillotine? AB - Since the discovery that the cysteine protease CED-3 was essential for developmental death in the nematode C. elegans, the search has been on to identify homologous proteases governing mammalian apoptosis. Fourteen of these proteases, now called caspases, have been found to date, and studies with natural or chemical inhibitors, and more recently knock-out mice, confirmed the involvement of at least a subset of these proteases in various forms of mammalian apoptosis. However, there has been recent evidence that some apoptotic morphologies, such as cell shrinkage, membrane blebbing and nuclear condensation, are not blocked by caspase inhibitors and that the cells continue to die in a protracted and inefficient manner. This has led to the notion that caspases are not required for all aspects of apoptosis in mammals. Here we review the current knowledge about caspase-independent apoptosis, discuss the strengths and weaknesses of the reasoning that led to its proposition and provide insights into its possible regulation and physiological significance. PMID- 10381653 TI - Caspase-independent programmed cell death with necrotic morphology. AB - Cell death is generally classified into two large categories: apoptosis represents active, programmed cell death, while necrosis represents passive cell death without underlying regulatory mechanisms. Recent progress revealed that caspases, a family of cysteine proteases, play a central role in the regulation of apoptosis. Unexpectedly, however, caspase inhibition occasionally turns the morphology of programmed cell death from apoptotic into necrotic without inhibiting death itself. In this article, we review different models of caspase independent programmed cell death showing necrotic-like morphology, including our Ras-mediated caspase-independent cell death. Based on these findings, we suggest the existence of a necrotic-like cell death regulated by cellular intrinsic death programs distinct from that of apoptosis. Even though type 2 physiological cell death, or autophagic degeneration, has been recognized as a necrotic-like programmed cell death for a long time, the underlying molecular mechanisms have not been identified despite its physiological significance. This has been in part due to the previous absence of adequate caspase-independent cellular models to study, recent efforts may now help to elucidate these mechanisms. PMID- 10381654 TI - Apoptosis inducing factor (AIF): a phylogenetically old, caspase-independent effector of cell death. AB - Although much emphasis has been laid on the role of caspase in cell death, recent data indicate that, in many instances, mammalian cell death is caspase independent. Thus, in many examples of mammalian cell death the 'decision' between death and life is upstream or independent of caspase activation. Similarly, it is unclear whether PCD of plants and fungi involves the activation of caspase-like enzymes, and no caspase-like gene has thus far been cloned in these phyla. Apoptosis inducing factor (AIF) is a new mammalian, caspase independent death effector which, upon apoptosis induction, translocates from its normal localization, the mitochondrial intermembrane space, to the nucleus. Once in the nucleus, AIF causes chromatin condensation and large scale DNA fragmentation to fragments of approximately 50 kbp. The AIF cDNA from mouse and man codes for a protein which possesses three domains (i) an amino-terminal presequence which is removed upon import into the intermembrane space of mitochondria; (ii) a spacer sequence of approximately 27 amino acids; and (iii) a carboxyterminal 484 amino acid oxidoreductase domain with strong homology to oxidoreductases from other vertebrates (X. laevis), non-vertebrate animals (C. elegans, D. melanogaster), plants, fungi, eubacteria, and archaebacteria. Functionally important amino acids involved in the interaction with the prosthetic groups flavin adenine nucleotide and nicotinamide adenine nucleotide are strongly conserved between AIF and bacterial oxidoreductase. Several eukaryotes possess a similar domain organisation in their AIF homologs, making them candidates to be mitochondrial oxidoreductases as well as caspase independent death effectors. The phylogenetic implications of these findings are discussed. PMID- 10381655 TI - NF-kappaB functions as both a proapoptotic and antiapoptotic regulatory factor within a single cell type. AB - Recently NF-kappaB has been shown to have both proapoptotic and antiapoptotic functions. In T cell hybridomas, both T cell activators and glucocorticoids induce apoptosis. Here we show that blockade of NF-kappaB activity, using a dominant negative IkappaBalpha, has opposite effects on these two apoptotic signals. Treatment with PMA plus ionomycin (P/I) results in the upregulation of Fas Ligand (FasL) and induction of apoptosis. Inhibition of NF-kappaB activity inhibits the P/I mediated induction of FasL mRNA and decreases the level of apoptosis in these cultures, thus establishing NF-kappaB as a proapoptotic factor in this context. Conversely, inhibition of NF-kappaB confers a tenfold increase in glucocorticoid mediated apoptosis, establishing that NF-kappaB also functions as an antiapoptotic factor. We conclude that NF-kappaB is a context-dependent apoptosis regulator. Our data suggests that NF-kappaB may function as an antiapoptotic factor in thymocytes while functioning as a proapoptotic factor in mature peripheral T cells. PMID- 10381656 TI - CD14 is a component of multiple recognition systems used by macrophages to phagocytose apoptotic lymphocytes. AB - Expression of the aminophospholipid phosphatidylserine (PS) on the surface of apoptotic lymphocytes and lipid-symmetric erythrocytes triggers their phagocytosis by macrophages. Phagocytosis by both activated and unactivated macrophages, which utilize different recognition systems, can be blocked by certain monoclonal antibodies directed against the LPS receptor, CD14. Here we investigate the requirement for CD14 in the phagocytosis of both apoptotic thymocytes and lipid-symmetric erythrocytes by both activated and unactivated macrophages. We show that phagocytosis of lipid-symmetric erythrocytes by both activated and unactivated macrophages is completely abolished when CD14 is removed from macrophages by cleaving its glycosylphosphatidylinositol tether with phospholipase C. This treatment also substantially reduces phagocytosis of apoptotic lymphocytes by both types of macrophages. Unactivated LR-9 mouse macrophages which are deficient in CD14 expression are completely unable to phagocytose either apoptotic thymocytes or lipid-symmetric erythrocytes. These results argue that CD14 is an absolute requirement for the phagocytosis of lipid symmetric erythrocytes by both activated and unactivated macrophages, despite their different recognition systems, that CD14 contributes at least substantially to the phagocytosis of apoptotic lymphocytes by both activated and unactivated macrophages, and that activated macrophages may also possess an alternate, CD14 independent mechanism for phagocytosis of apoptotic lymphocytes. PMID- 10381658 TI - Writing to our patients. PMID- 10381657 TI - A role for hepatocyte growth factor in diabetic retinopathy? PMID- 10381659 TI - New treatment of dry eye: the effect of calcium ointment through eyelid skin delivery. AB - AIM: To demonstrate the efficacy of a petrolatum based calcium ointment applied to the lower lid skin in the management of dry eye. METHODS: In a controlled double masked study, the effects of water free petrolatum ointment containing calcium carbonate (10% w/w) on tear functional factors and ocular surface vital staining in dry eye patients were observed. Petrolatum without calcium carbonate served as control. Patients were instructed to place ointment to the lower lid skin twice a day. Evaluation of subjective complaints, fluorescein and rose bengal staining patterns, blink rate, tear evaporation and tear break up time (BUT) were performed before and 3 months after treatment. In order to demonstrate the movement of petrolatum from the skin to the tear film, petrolatum containing 1% sodium fluorescein was placed on the lower lid of four healthy volunteers, and the concentration of fluorescein in the tear film was followed up to 6 hours using an anterior fluorometer. RESULTS: Subjective symptoms significantly improved in both the calcium group (p=0.001) and control (p=0.012), while only the calcium group demonstrated a significant improvement in fluorescein (p=0.043), rose bengal (p=0.021) scores, and blink rate (p=0.004). Tear evaporation also significantly decreased in both the calcium group (p=0.0004) and control (0.043). BUT did not improve in either group. CONCLUSION: Petrolatum based calcium ointment significantly improved symptoms, tear dynamics, and ocular surface staining in dry eye patients. However, some of the therapeutic effects may be due to lipids in the petrolatum vehicle. Petrolatum applied to the lower lid skin is an effective drug delivery system for slowly releasing drugs to the ocular surface. PMID- 10381660 TI - Identification and antibiotic susceptibility of coagulase negative staphylococci isolated in corneal/external infections. AB - AIMS: To identify and determine antibiotic susceptibility of coagulase negative staphylococci (CoNS) isolated from patients with chronic blepharitis, purulent conjunctivitis, and suppurative keratitis. METHODS: A retrospective review of all culture positive cases of chronic blepharitis, purulent conjunctivitis, and suppurative keratitis between July 1995 and December 1996 was performed. Cases in which CoNS were the sole isolates were analysed. Species identification was performed by using a commercially available standardised biochemical test system. Antibiotic susceptibility to penicillin, gentamicin, tetracycline, erythromycin, ciprofloxacin, and teicoplanin was determined by agar disc diffusion (Kirby-Bauer method). Teicoplanin resistance was confirmed by agar dilution. RESULTS: 42 Staphylococcus epidermidis, four S warneri, three S capitis, two S hominis, one each of S xylosus, S simulans, S equorum, and S lugdunensis were identified. 37 CoNS were penicillin resistant, 12 gentamicin resistant, 28 tetracycline resistant, 18 erythromycin resistant, four ciprofloxacin resistant, and one teicoplanin resistant (MIC, 32 microg/ml). In total, 16 strains were resistant to three or more antibiotics. CONCLUSION: Species of CoNS apart from S epidermidis may be isolated from patients with corneal and external infection. Antibiotic susceptibility of CoNS is unpredictable and multiresistant strains are common. As a result, antibiotic susceptibility testing should be performed in all cases of clinically significant ocular infections caused by CoNS. PMID- 10381661 TI - Evaluation of corneal thickness and topography in normal eyes using the Orbscan corneal topography system. AB - AIMS: To map the thickness, elevation (anterior and posterior corneal surface), and axial curvature of the cornea in normal eyes with the Orbscan corneal topography system. METHODS: 94 eyes of 51 normal subjects were investigated using the Orbscan corneal topography system. The anterior and posterior corneal elevation maps were classified into regular ridge, irregular ridge, incomplete ridge, island, and unclassified patterns, and the axial power maps were grouped into round, oval, symmetric bow tie, asymmetric bow tie, and irregular patterns. The pachymetry patterns were designated as round, oval, decentred round, and decentred oval. RESULTS: The thinnest point on the cornea was located at an average of 0.90 (SD 0. 51) mm from visual axis and had an average thickness of 0.55 (0.03) mm. In 69.57% of eyes, this point was located in the inferotemporal quadrant, followed by the superotemporal quadrant in 23.91%, the inferonasal quadrant in 4.35%, and the superonasal quadrant in 2.17%. Among the nine regions of the cornea evaluated (central, superotemporal, temporal, inferotemporal, inferior, inferonasal, nasal, superonasal, and superior) the central cornea had the lowest average thickness (0.56 (0.03) mm) and the superior cornea had the greatest average thickness (0.64 (0.03) mm). The mean simulated keratometry (SimK) was 44.24 (1.61)/43.31 (1.66) dioptres (D) and the mean astigmatism was 0.90 (0.41) D. Island (71.74%) was the most common elevation pattern observed in the anterior corneal surface, followed by incomplete ridge (19.57%), regular ridge (4.34%), irregular ridge (2.17%), and unclassified (2.17%). Island (32.61%) was the most common topographic pattern in the posterior corneal surface, following by regular ridge (30.43%), incomplete ridge (23. 91%), and irregular ridge (13.04%) patterns. Symmetric bow tie was the most common axial power pattern in the anterior cornea (39.13%), followed by oval (26.07%), asymmetric bow tie (23.91%), round (6. 52%), and irregular (4.53%) patterns. In the pachymetry maps, 47.83% of eyes had an oval pattern, and round, decentred oval, and decentred round were observed in 41.30%, 8.70%, and 2.18% of eyes, respectively. CONCLUSION: The information on regional corneal thickness, corneal elevation and axial corneal curvature obtained with the Orbscan corneal topography system from normal eyes provides a reference for comparison with diseased corneas. The Orbscan corneal topography system is a useful tool to evaluate both corneal topography and corneal thickness. PMID- 10381662 TI - Topographic and keratometric astigmatism up to 1 year following small flap trabeculectomy (microtrabeculectomy). AB - AIM: To determine the induced corneal astigmatism by measuring the changes in manual keratometry and computerised corneal videokeratoscopy up to 1 year following small flap trabeculectomy (microtrabeculectomy). METHOD: A prospective study of a case series of small flap trabeculectomy procedures performed at the 90 degree meridian on 16 eyes of 16 patients, all followed to 1 year postoperatively. Changes in manual keratometry and computerised videokeratoscopy (Eyesys) readings were analysed by vector analysis and vector decomposition techniques. RESULTS: By vector analysis, the mean surgically induced refractive change (SIRC) cylinder power vectors induced at 1, 3, 6, and 12 months as measured by manual keratometry were 0.68, 0.38, 0.52, and 0.55 dioptres, and by keratography 0.75, 0.66, 0.59, and 0.64 dioptres. Vector decomposition on the induced vector cylinders on manual keratometry resulted in a "with the rule" mean vector of 0.52 and 0.22 dioptres at 1 and 3 months and an "against the rule" mean vector of 0.16 and 0.16 dioptres at the same time points (p=0.03 and 0.28 respectively). Vector decomposition at 6 and 12 months revealed no significant with the rule changes induced. Similar analysis on the videokeratoscopy results revealed significant induced with the rule astigmatism until 3 months, but not at 6 and 12 months postoperatively. CONCLUSION: Small flap trabeculectomy (microtrabeculectomy) produces smaller changes in corneal curvature that resolve sooner than previous reports of larger flap techniques. PMID- 10381663 TI - Early retreatment of infantile esotropia: comparison of reoperation and botulinum toxin. AB - AIM: To compare the efficacy of reoperation and botulinum toxin injection in treating infantile esotropes early after unsatisfactory surgical alignment. METHODS: 55 strabismic children who had been unsuccessfully operated for infantile esotropia were randomised to reoperation (28 patients) or botulinum toxin injection (27 patients). The motor outcomes (percentage of successful motor outcome and percentage change in deviation) were compared at 6 months, 1 year, and 3 years after retreatment, and the sensory outcomes (percentage with fusion ability and stereo perception) at the 3 year follow up visit. RESULTS: The motor and sensory outcomes and the stability of motor results were similar in patients reoperated and treated with botulinum injection. At the 3 year visit 67.8% and 59.2% of children were, respectively, within 8 prism dioptres of orthotropia (p=0.72). The frequency of fusion ability was, respectively, 60.7% and 51.8% (p=0.71), and the frequency of stereo perception (100 microM at hmGluR1. Schild analysis demonstrated a noncompetitive mechanism of inhibition. Pharmacophore mapping was used to identify an additional compound, SIB-1893 [(E)-2-methyl-6-(2 phenylethenyl)pyridine], which was also shown to block glutamate-induced increases in [Ca2+]i at hmGluR5 with an IC50 of 0.29 microM compared with an IC50 of >100 microM at hmGluR1. SIB-1757 and SIB-1893 showed little or no activity when tested for agonist and antagonist activity at the other recombinant human mGluR subtypes, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid, kainate, and N-methyl-D-aspartate receptors. In rat neonatal brain slices, SIB 1757 and SIB-1893 inhibited (S)-3,5-dihydroxyphenylglycine (DHPG)-evoked inositol phosphate accumulation in hippocampus and striatum by 60% to 80%, with a potency similar to that observed on recombinant mGluR5. However, in the cerebellum, a brain region with low mGluR5 expression, SIB-1757 failed to inhibit DHPG-evoked inositol phosphate accumulation. In cultured rat cortical neurons, SIB-1757 and SIB-1893 largely inhibited DHPG-evoked [Ca2+]i signals, revealing a population of neurons that were less sensitive to SIB-1757 and SIB-1893. This is the first description of highly selective, noncompetitive mGluR5 antagonists. These compounds will be useful tools in evaluating the role of mGluR5 in normal physiology and in animal models of disease. PMID- 10381774 TI - Prostate-specific human N-acetyltransferase 2 (NAT2) expression in the mouse. AB - 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is a heterocyclic amine identified in the human diet and in cigarette smoke that produces prostate tumors in the rat. PhIP is bioactivated by cytochrome P-450 enzymes to N-hydroxylated metabolites that undergo further activation by conjugation enzymes, including the N-acetyltransferases, NAT1 and NAT2. To investigate the role of prostate-specific expression of human N-acetyltransferase 2 (NAT2) on PhIP-induced prostate cancer, we constructed a transgenic mouse model that targeted expression of human NAT2 to the prostate. Following construction, prostate, liver, lung, colon, small intestine, urinary bladder, and kidney cytosols were tested for human NAT1- and NAT2-specific N-acetyltransferase activities. Human NAT2-specific N acetyltransferase activities were 15-fold higher in prostate of transgenic mice versus control mice, but were equivalent between transgenic mice and control mice in all other tissues tested. Human NAT1-specific N-acetyltransferase activities did not differ between transgenic and control mice in any tissue tested. Prostate cytosols from transgenic and control mice did not differ in their capacity to catalyze the N-acetylation of 2-aminofluorene, the O-acetylation of N-hydroxy-2 aminofluorene and N-hydroxy-PhIP or the N,O-acetylation of N-hydroxy-2 acetylaminofluorene. Transgenic and control mice administered PhIP did not differ in PhIP-DNA adduct levels in the prostate. This study is the first to report transgenic expression of human NAT2 in the mouse. The results do not support a critical role for bioactivation of heterocyclic amine carcinogens by human N acetyltransferase-2 in the prostate. However, the lack of an effect may relate to the level of overexpression achieved and the presence of endogenous mouse acetyltransferases and/or sulfotransferases. PMID- 10381776 TI - Effect of chronic social stress on delta-opioid receptor function in the rat. AB - Previous studies have shown that stressors modify endogenous opioid systems. However, the consequences of social stress on the function of endogenous opioid systems is not well documented. The present studies investigated the effect of rank and housing condition on response to SNC-80, a delta receptor agonist. Triad housed rats were assessed for dominance status by their behavior and alteration in body weights. At 3 and 50 days, triad- and individually housed rats were injected with SNC-80 (35 mg/kg i.p.) or saline, and evaluated using a test battery consisting of open field behaviors, rectal temperature, analgesia, and air-puff-induced ultrasonic vocalizations. After 50 days of housing, plasma corticosterone, adrenal catecholamines, and the density of cyclic[D penicillamine2-D-penicillamine2]enkephalin-stimu lat ed guanylyl 5' [gamma[35S]thio]-triphosphate binding in the prefrontal cortex, the amygdala, nucleus accumbens, thalamus, arcuate, and median eminence were also determined. The first 24 h of triad housing resulted in loss of body weight in subdominant (betas and gammas) but not dominant alpha rats. SCN-80-induced hypothermia was smaller, and there was no depression of headpoke and locomotor behavior in the periphery and the center of the field of alpha rats, in contrast to subdominant and singly housed rats. Rank status did not influence SNC-80's analgesic effect or its inhibition of air-puff-induced ultrasonic vocalizations. Plasma corticosterone levels of alphas and gammas were lower compared with betas and singly housed rats. Agonist stimulation of delta receptor guanylyl 5' [gamma[35S]thio]-triphosphate binding was lateralized in prefrontal cortex and amygdala, but not nucleus accumbens. Binding was highest in all brain areas of singly housed rats and lowest in the thalamus of beta and of gamma rats. Lateralized binding in amygdala, high locomotor activity, and sensory sensitivity correlated positively with greater sensitivity to SNC-80-induced depression in these measures. Higher binding in the right amygdala correlated with higher plasma corticosterone levels. These findings indicate that dominant rats displayed stimulant rather than depressant responses to delta-opioid activation. Therefore in rodents rank-related stress can alter responsiveness of the endogenous opioid system, and dominance can increase the excitatory effects of delta agonists. PMID- 10381775 TI - Preclinical evaluation of anti-inflammatory activities of the novel pyrrolopyrimidine PNU-142731A, a potential treatment for asthma. AB - The anti-inflammatory properties of a novel pyrrolopyrimidine, PNU-142731A, in a murine model of antigen-induced eosinophilic lung inflammation are described. PNU 142731A, when given orally, demonstrated a dose-related inhibition of eosinophil- and lymphocyte-rich accumulation in the airways of ovalbumin (OA)-sensitized and challenged (OA/OA) C57BL/6 mice. The magnitude of the suppression of lung inflammation was also dependent on length of treatment. Reductions in the levels of interleukin (IL)-5, IL-6, and IgA in the bronchoalveolar lavage fluid of treated OA/OA mice, when compared with vehicle-sensitized control mice (V/OA), were observed. Plasma concentrations of IL-5, total IgE, and OA-specific IgG1 were also lowered in OA/OA mice by treatment. Histological assessment of formalin fixed lung tissue sections confirmed that the compound blocked the accumulation of eosinophils in the airway tissue. Furthermore, significantly less mucus glycoproteins were seen in the lungs of PNU-142731A-treated OA/OA mice. Reverse transcription-polymerase chain reaction of lung tissue from PNU-142731A-dosed OA/OA mice demonstrated that mRNA for Th2 cytokines was less than that in vehicle treated OA/OA controls. OA-elicited production of IL-4 by disaggregated lung tissue cells from PNU-142371A-treated OA/OA mice was also less than that of controls. In contrast, the release of Th1 cytokines (IL-2 and interferon-gamma) were elevated. Similarly, the OA-stimulated release of Th2 cytokines (IL-5 and IL 10) by splenocytes from PNU-142731A-treated OA/OA mice were inhibited. Combined therapy of OA/OA mice with PNU-142731A and suboptimal doses of dexamethasone revealed that PNU-142731A had steroid-sparing effects. These characteristics of PNU-142731A in a murine model of allergic tissue inflammation support its clinical development as a potential treatment for asthma. PMID- 10381777 TI - Biphasic modulation of visceral nociception by neurotensin in rat rostral ventromedial medulla. AB - A potential role for neurotensin in the rostral ventromedial medulla (RVM) in modulation of visceral nociceptive transmission was examined in this study. Microinjection of neurotensin (3-3000 pmol) into the RVM of awake rats produced a dose-dependent inhibition of the visceromotor response (VMR) to noxious colorectal distension (CRD) that lasted 30 to 120 min. Additionally, intra-RVM injection of neurotensin (300 pmol) significantly reduced the slope of the stimulus-response function to graded CRD (20-80 mm Hg), whereas the greatest dose of neurotensin (3000 pmol) completely inhibited the VMR at all intensities of CRD. General motor function was unaffected after intra-RVM injection of neurotensin (3000 pmol). Intra-RVM injection of lesser doses of neurotensin (0.03 0.30 pmol) resulted an enhancement of the VMR to noxious CRD that had a short duration (18-30 min), and produced a leftward shift of the stimulus-response function to graded CRD without a change in the slope of the function. Additionally, intra-RVM injection of the neurotensin-receptor antagonist SR48692 (0.3-300 fmol) in naive animals produced dose-dependent inhibition of VMR to noxious CRD, whereas a lesser dose (0.03 fmol) enhanced the VMR. These data support a role for neurotensin in the RVM in biphasic modulation of visceral nociception. The results obtained with SR48692 suggest that endogenous neurotensin in the RVM modulates VMR to noxious CRD via a prominent interaction with neurotensin receptors that mediate facilitatory influences and a lesser interaction with neurotensin receptors that mediate masked inhibitory influences. PMID- 10381778 TI - Gabapentin attenuates nociceptive behaviors in an acute arthritis model in rats. AB - In this study, we investigated the effectiveness of gabapentin (Neurontin), administered spinally with a microdialysis fiber, in reducing nociceptive behavioral responses induced by a knee joint inflammation model. This model is produced by injection of the knee joint with kaolin and carrageenan in rats. The resultant knee joint inflammation produces a secondary hyperalgesia to radiant heat applied to the hindpaw. Both pretreatment and post-treatment protocols were examined. Spinal administration of gabapentin (10 mg/ml) infused 1.5 h before induction of knee joint inflammation, although having no effect on the baseline, prevented the development of heat hyperalgesia. Gabapentin also prevented the development of other pain-related behaviors scored subjectively. Gabapentin had no effect, however, on the joint circumference increase typical in this model. In animals with fully developed knee joint inflammation, gabapentin produced a reversal of heat hyperalgesia. The paw withdrawal latency responses and subjective pain scores were no longer significantly different from baseline, but joint circumference increases remained. These data suggest that gabapentin is an effective antinociceptive agent when administered either before or after induction of knee joint inflammation acting through a central neurogenic mechanism. PMID- 10381779 TI - AA-2414, an antioxidant and thromboxane receptor blocker, completely inhibits peroxide-induced vasoconstriction in the human placenta. AB - We hypothesized that AA-2414, a novel thromboxane receptor blocker with antioxidant properties, would inhibit peroxide-induced vasoconstriction in the isolated perfused human placental cotyledon. In study 1, placental cotyledons (n = 5) were perfused serially for 20- min intervals with control KrebsRinger bicarbonate (KRB) buffer, t-butyl hydroperoxide (Px; 100 microM), KRB buffer, and KRB buffer containing Px to which progressively increasing concentrations of AA 2414 were added (1 x 10(-8) to 1 x 10(-4) mol/l). In study 2, placental cotyledons (n = 6) were perfused with control KRB buffer, Px alone, KRB buffer, 1 x 10(-5) mol/l AA-2414 alone, Px plus AA-2414, and Px alone. Compared with control, perfusion with Px significantly increased perfusion pressure, vascular resistance, and the maternal and fetal secretion rates of lipid peroxides, thromboxane B2 (TXB2) and 6-keto prostaglandin F1alpha. In study 1, AA-2414 + Px produced a dose-response inhibition of Px-induced increases in perfusion pressure, vascular resistance, and maternal secretion of lipid peroxides and TXB2. In study 2, perfusing AA-2414 at a dose of 1 x 10(-5) mol/l completely inhibited Px-induced vasoconstriction and increases in lipid peroxide and TXB2 secretion rates, but only partially inhibited the increase in 6-keto prostaglandin F1alpha secretion. We conclude that AA-2414 inhibited peroxide induced vasoconstriction in the human placenta, as well as peroxide- induced increases in the placental secretion rates of lipid peroxides and thromboxane, but only partially inhibited peroxide-induced increases in the placental secretion rate of prostacyclin. PMID- 10381780 TI - Interactions of 6-gingerol and ellagic acid with the cardiac sarcoplasmic reticulum Ca2+-ATPase. AB - The inotropic/lusitropic effects of beta-adrenergic agonists on the heart are mediated largely by protein kinase A (PKA)-catalyzed phosphorylation of phospholamban, the natural protein regulator of the Ca2+ pump present in sarcoplasmic reticulum (SR) membranes. Gingerol, a plant derivative, is known to produce similar effects when tested in isolated cardiac muscle. The purpose of the present study was to compare the effects of gingerol and another plant derivative, ellagic acid, on the kinetics of the SR Ca2+ pump with those of PKA catalyzed phospholamban phosphorylation to elucidate their mechanisms of Ca2+ pump regulation. As previously demonstrated for PKA, 50 microM gingerol or ellagic acid increased Vmax(Ca) of Ca2+ uptake and Ca2+-ATPase activity assayed at millimolar ATP concentrations in light cardiac SR vesicles. Unlike PKA, which decreases Km(Ca), neither compound had a significant effect on Km(Ca) in unphosphorylated vesicles. However, gingerol increased Km(Ca) in phosphorylated vesicles, in which Ca2+ uptake was significantly increased further at saturating Ca2+ and remained unchanged at subsaturating Ca2+. An inhibition of Ca2+ uptake by gingerol at micromolar MgATP concentrations was overcome with increasing MgATP concentrations. The stimulation of Ca2+ uptake attributable to gingerol in unphosphorylated microsomes at saturating Ca2+ was 30% to 40% when assayed at 0.05 to 2 mM MgATP and only about 12% in phosphorylated microsomes as well as in rabbit fast skeletal muscle light SR. The present results support the view that an ATP-dependent increase in Vmax(Ca) of the SR Ca2+ pump plays an important role in mediating cardiac contractile responses to gingerol and phospholamban dependent beta-adrenergic stimulation. PMID- 10381781 TI - Reperfusion injury in livers due to gentle in situ organ manipulation during harvest involves hypoxia and free radicals. AB - Kupffer cell-dependent injury in livers gently manipulated during harvest develops upon transplantation; however, underlying mechanisms remain unknown. Thus, the purpose of this study was to identify factors involved in mechanisms of injury. Livers from female Sprague-Dawley rats (200-230 g) were cold stored for 24 h in University of Wisconsin solution. Subsequently, livers were perfused at 37 degrees C with oxygen-saturated Krebs-Henseleit buffer containing fluorescein dextran to assess microcirculation. Cell death was assessed by uptake of trypan blue, a vital dye. Minimal dissection during harvest had no effects on sinusoidal lining cells; however, gentle organ manipulation dramatically increased trypan blue uptake about 5-fold (p <.05). In contrast, perfusion with N2-saturated buffer after cold storage totally prevented cell death due to manipulation. At harvest, portal venous pressure was increased significantly by 70% due to manipulation. Furthermore, vascular space and microcirculation were decreased by more than 50% (p <.05), reflecting the rate of entry and exit of fluorescein dextran. Pimonidazole, a 2-nitroimidazole marker, was given to rats before harvest to detect hypoxia in liver. Pimonidazole adduct binding was increased significantly about 2-fold by manipulation. To detect free radical adducts by electron spin resonance (ESR) spectroscopy in bile, C-phenyl-N-tert-butylnitrone was given as spin trapping reagent to the donor before operation. Free radical formation was increased about 3-fold by organ manipulation (p <.05). Donors given gadolinium chloride, a selective Kupffer cell toxicant, or dietary glycine, which prevents activation of Kupffer cells, significantly blunted microcirculatory disturbances, hypoxia, and death of endothelial lining cells. These data indicate for the first time that gentle organ manipulation during harvest causes oxygen dependent reperfusion injury to endothelial lining cells via mechanisms involving hepatic microcirculation, hypoxia, and Kupffer cells. PMID- 10381782 TI - Differential cotransmission in sympathetic nerves: role of frequency of stimulation and prejunctional autoreceptors. AB - Recent reports have suggested that sympathetic nerves may store separately and release independently the cotransmitters ATP and norepinephrine (NE). It is conceivable therefore that the quantity of each neurotransmitter that is released from the nerves is not fixed but rather may vary, possibly with the frequency of stimulation. To test this hypothesis we studied the concomitant release at various frequencies and cooperative postjunctional actions of ATP and NE during the first 10 s of electrical field stimulation of the guinea pig vas deferens. We found that at lower frequencies (8 Hz), prejunctional inhibition of the release of NE, which occurs via alpha2-adrenoceptors, modulates the ultimate composition of the cocktail of cotransmitters by limiting the amount of NE that is coreleased with ATP. As the frequency of stimulation increases (above 8 Hz), the autoinhibition of the release of NE is overridden and the amount of NE relative to ATP increases. The smooth muscle of the guinea pig vas deferens reacts to changes in composition of the sympathetic neurochemical messages by increasing the amplitude of its contractions due to the enhancement by NE of the contractile responses triggered by ATP. This evidence suggests that the prejunctional alpha2 adrenoceptor may function as a sensor that "reads" the frequency of action potentials produced during a burst of neuronal activity and converts that information into discrete neurochemical messages with varying proportions of cotransmitters. The mechanism for decoding the informational content of these messages is based on the cooperative postjunctional interactions of the participating cotransmitters. PMID- 10381783 TI - Topical opioids in mice: analgesia and reversal of tolerance by a topical N methyl-D-aspartate antagonist. AB - In addition to its central actions, morphine has important peripheral effects. To examine peripheral analgesic mechanisms, we developed a topical opioid paradigm in which the tail was immersed in a dimethyl sulfoxide (DMSO) solution containing various drugs. Alone, DMSO was inactive in the tail-flick assay in mice. DMSO solutions containing morphine and peptides such as [D-Ala2,MePhe4, Gly(ol)5]enkephalin (DAMGO) produced a potent, dose-dependent analgesia with the radiant heat tail-flick assay. The actions of the drugs were local. Analgesia was observed only in regions of the tail exposed to the solution and not in more proximal unexposed portions of the tail. Immersion of the tail in a solution containing either 125I-labeled morphine or 125I-labeled DAMGO revealed no detectable uptake of radioactivity into the brain, spinal cord, or blood. In the tail, radioactivity was limited only to the regions actually immersed in the solutions. The topical drugs potentiated systemic agents, similar to the previously established synergy between peripheral and central sites of action. Local tolerance was rapidly produced by repeated daily exposure of the tail to morphine. Topical morphine tolerance was effectively blocked by the N-methyl-D aspartate (NMDA) antagonist MK801 given either systemically or topically but not intrathecally. The ability of a topical NMDA antagonist to block local morphine tolerance suggests that peripheral NMDA receptors mediate topical morphine tolerance. Morphine was cross-tolerant to DAMGO, but not to morphine-6beta glucuronide, implying different mechanisms of action. These observations are significant in the design and use of opioids clinically. PMID- 10381784 TI - Modulation of effect of dietary salt on prehepatic first-pass metabolism: effects of beta-blockade and intravenous salt loading. AB - We previously demonstrated that increased dietary salt markedly decreases plasma quinidine concentrations shortly after p.o. dosing, without an effect on the drug's terminal elimination half-life or concentrations after i.v. administration. These findings suggest an effect of dietary salt on intestinal metabolism or transport of the drug. Because one effect of salt loading is sympathetic inhibition, we examined the effect of beta-adrenoceptor blockade on salt-related changes in quinidine disposition. Furthermore, we examined whether the action of salt is local or systemic by determining the effect of salt loading by the i.v. route. To assess the effect of beta-blockade, quinidine disposition was studied in eight normal volunteers after a single p.o. dose of quinidine; data were obtained after 1 week on a high-salt diet (400 mEq/day) and 1 week on a low-salt diet (10 mEq/day) during chronic nadolol and compared with those previously obtained in the same subjects without the beta-blocker. beta-Blockade had no effect on oral clearance during the high-salt diet [0.28 +/- 0.1 (quinidine + nadolol) versus 0.30 +/- 0.2 liters/h/kg (quinidine alone)] but increased clearance on the low-salt diet from 0.23 +/- 0.1 to 0.29 +/- 0.1 liters/h/kg (p <. 05). For the i.v. salt study, the disposition of single p.o. and single i.v. doses of quinidine was determined on two occasions in eight subjects: once during a low-salt diet (10 mEq/day) and once during the same diet, supplemented by 400 mEq/day NaCl i.v. for 8 days. In contrast to our findings after p.o. salt loading, i.v. salt loading did not alter the pharmacokinetics of p.o. quinidine. Taken together, these data implicate a local alteration of drug metabolizing activity and/or drug transport in the intestinal mucosa as the major effect of dietary salt on the disposition of p.o. quinidine and further suggest that beta-adrenergic activation by a low-salt diet is one component of a signaling pathway whereby intestinal drug disposition is suppressed, resulting in increased oral bioavailability. PMID- 10381786 TI - Dual pharmacological properties of a cyclic AMP-sensitive potassium channel. AB - Bovine adrenal zona fasciculata cells express a novel K+ current (IAC) that sets the resting potential while it couples adrenocorticotropin and angiotensin II receptors to membrane depolarization and cortisol secretion. IAC is distinctive among K+ channels both in its activation by ATP and its inhibition by cyclic AMP. Whole-cell and single-channel patch-clamp recording was used to establish a pharmacological profile of IAC K+ channels. IAC was blocked by antagonists of cyclic nucleotide-gated channels, including the diphenylbutylpiperidine (DPBP) antipsychotic pimozide and l-cis-diltiazem. Other DPBPs, including penfluridol and fluspirilene, also potently inhibited this channel. The inhibition of IAC by DPBPs was selective because 200-fold higher concentrations of penfluridol were required to inhibit voltage-gated IA K+ channels in adrenal zona fasciculata cells. Standard K+ channel antagonists blocked IAC at concentrations 100- to 100,000-fold higher than the DPBPs. IAC channels were also inhibited by the sulfonylureas glyburide and tolbutamide but at concentrations higher than those that typically block ATP-sensitive inward rectifier K+ channels. Overall, the relative order of potency and associated IC50 values for IAC antagonists were as follows: penfluridol (0.187 microM) > fluspirilene (0.232 microM) > pimozide (0.354 microM) >> l-cis-diltiazem (24.9 microM) approximately quinidine (24.1 microM) > bupivacaine (113.2 microM) > tolbutamide (784.4 microM) > BaCl2 (1027 microM) > 4-aminopyridine (2750 microM) > tetraethylammonium (24,270 microM). IAC channels are unique in combining the pharmacological properties of K+-selective channels with those of cyclic nucleotide-gated cation channels. The potent block of IAC channels identifies DPBPs as a new class of K+ channel antagonists and suggests additional targets for these neuroleptics in the central nervous system. PMID- 10381785 TI - kappa-Opioid receptor effects of butorphanol in rhesus monkeys. AB - Butorphanol and nalbuphine have substantial affinity for mu and kappa-opioid receptor sites, yet their behavioral effects in monkeys are largely consistent with a mu receptor mechanism of action. Using ethylketocyclazocine (EKC) discrimination and diuresis assays in rhesus monkeys (Macaca mulatta), the purpose of the current investigation was to characterize the in vivo kappa-opioid activity of these compounds through the use of an insurmountable mu-opioid receptor antagonist, clocinnamox. Alone, butorphanol (0.001-0.032 mg/kg i.m.) failed to generalize to EKC, and pretreatment with the competitive opioid receptor antagonist quadazocine (0.1 or 0.32 mg/kg i.m.) did not alter this generalization. At 24 h after clocinnamox (0.1 mg/kg i.m.) administration, butorphanol fully generalized to EKC, and this generalization was maintained in two of three monkeys at 72 h. Parallel results were observed in diuresis: butorphanol alone and in the presence of quadazocine (1 mg/kg i.m.) did not alter urine output, and a marked diuretic effect was demonstrated 24 h to 2 weeks after clocinnamox administration. Clocinnamox did not alter the discriminative stimulus or diuretic effects of nalbuphine or of the kappa-opioid receptor agonists EKC or U69593. These results are consistent with an in vivo agonist activity of butorphanol at kappa-opioid receptors that can only be demonstrated when an insurmountable antagonist has substantially eliminated the dominant receptor population through which it exerts its action. PMID- 10381787 TI - Dose-dependent inhibition of platelet cyclooxygenase-1 and monocyte cyclooxygenase-2 by meloxicam in healthy subjects. AB - We evaluated whether therapeutic blood levels of meloxicam are associated with selective inhibition of monocyte cyclooxygenase (COX)-2 in vitro and ex vivo. Concentration-response curves for the inhibition of monocyte COX-2 and platelet COX-1 were obtained in vitro after the incubation of meloxicam with whole blood samples. Moreover, 11 healthy volunteers received placebo or 7.5 or 15 mg/day meloxicam, each treatment for 7 consecutive days, according to a randomized, double-blind, crossover design. Before dosing and 24 h after the seventh dose of each regimen, heparinized whole blood samples were incubated with lipopolysaccharide (10 microgram/ml) for 24 h at 37 degrees C, and prostaglandin E2 was measured in plasma as an index of monocyte COX-2 activity. The production of thromboxane B2 in whole blood allowed to clot at 37 degrees C for 60 min was assessed as an index of platelet COX-1 activity. The administration of placebo did not significantly affect plasma prostaglandin E2 (21. 3 +/- 7.5 versus 19.1 +/- 4 ng/ml, mean +/- S.D., n = 11) or serum thromboxane B2 (426 +/- 167 versus 425 +/- 150 ng/ml) levels. In contrast, the administration of 7.5 and 15 mg of meloxicam caused dose-dependent reductions in monocyte COX-2 activity by 51% and 70%, respectively, and in platelet COX-1 activity by 25% and 35%, respectively. Although the IC50 value of meloxicam for inhibition of COX-1 was 10-fold higher than the IC50 value of COX-2 in vitro, this biochemical selectivity was inadequate to clearly separate the effects of meloxicam on the two isozymes after oral dosing as a function of the daily dose and interindividual variation in steady-state plasma levels. PMID- 10381788 TI - Role of murine cytochrome P-450 2F2 in metabolic activation of naphthalene and metabolism of other xenobiotics. AB - Despite their substantially lower levels relative to hepatic tissue, pulmonary cytochrome P-450 (CYP) monooxygenases play an important role in the metabolic activation of substrates that cause lung injury. The target- and species selective toxicity of a number of pulmonary toxicants has been attributed to the presence and distribution of activating enzymes with high kcat in target airways of susceptible species. However, experimental demonstration of these concepts and quantitative assessment of the contribution of individual CYP isoforms is lacking. This study was undertaken to characterize the catalytic activities of CYP2F2 with naphthalene, a murine Clara cell toxicant, as well as with other xenobiotics that either undergo metabolic activation to cytotoxic intermediates or that function as "isoform-selective" substrates. Recombinant CYP2F2 was produced using the baculovirus expression vector system in Spodoptera frugiperda and Trichoplusia ni cells, accounting up to approximately 20% of the total cellular protein. Incubations containing naphthalene, recombinant CYP2F2, NADPH cytochrome P-450 oxidoreductase, and NADPH-regenerating system metabolized naphthalene with a high degree of stereoselectivity to 1R, 2S-naphthalene oxide (66:1 enantiomeric ratio). The Km and kcat values, along with the specificity constant, for naphthalene metabolism by recombinant CYP2F2 were 3 microM, 104 min 1, and 5.8 x 10(5) M-1 s-1, respectively. Recombinant CYP2F2 also metabolized ethoxyresorufin, pentoxyresorufin, p-nitrophenol, and 1-nitronaphthalene at easily detectable levels. The results from this work suggest that CYP2F2 1) plays a key role in the species- and cell-selective toxicity of naphthalene and 2) efficiently metabolizes a number of other substrates, including the lung toxicant 1-nitronaphthalene. PMID- 10381789 TI - Transepithelial transport of organic anions across the choroid plexus: possible involvement of organic anion transporter and multidrug resistance-associated protein. AB - Transport characteristics of 17beta-estradiol 17beta-D-glucuronide (E217betaG), a dual substrate of the transporters for cellular uptake (organic anion transporting polypeptide 1 or oatp1) and cellular excretion (multidrug resistance associated protein 1or MRP1), in the rat choroid plexus were studied in vivo and in vitro. The uptake of E217betaG into isolated choroid plexus was mediated by an energy-dependent system with a Km of 3.4 microM. Together with the previous finding that oatp1 is localized on the apical membrane of choroid plexus, these results suggest that oatp1 is responsible for the uptake of this ligand. After intracerebroventricular administration, elimination of E217betaG from cerebrospinal fluid was probenecid sensitive and much more rapid than that of inulin; less than 2% of the administered E217betaG and 40 to 50% of inulin remained in the cerebrospinal fluid 20 min after intracerebroventricular administration. In addition, the amount of E217betaG associated with choroid plexus at 20 min was negligible, suggesting the presence of an efficient excretion system on the basolateral membrane of choroid plexus. Expression of MRP1 was detected in choroid plexus. Semiquantitative reverse transcription polymerase chain reaction and Western blot analyses indicated that the expression level of MRP1 in choroid plexus is about four or five times higher than that in the lung, one of the tissues exhibiting high expression of MRP1. Together with the in vivo vectorial transport of E217betaG, these results can be accounted for by assuming that there is basolateral localization of MRP1 in choroid plexus. Combined, oatp1 and MRP1 may synergistically mediate the efficient transcellular transport of E217betaG across choroid plexus. PMID- 10381790 TI - Tetraethylammonium and amantadine identify distinct organic cation transporters in rat renal cortical proximal and distal tubules. AB - Tetraethylammonium (TEA) and amantadine are two organic cations that are secreted by the kidney. It appears that each cation may characterize distinct renal tubule organic cation transport pathways. To test this hypothesis, we investigated the renal proximal and distal tubule energy-dependent transport properties of TEA and amantadine. Isolated tubules were incubated at 25 degrees C in bicarbonate buffer (Krebs-Henseleit solution) and nonbicarbonate buffer (Cross-Taggart) with varying concentrations of [14C]TEA or [3H]amantadine to determine initial rates of energy dependent uptake of TEA and amantadine, respectively. The uptake of TEA could best be described by two transport sites, a high-affinity site and a lower affinity site. TEA uptake was not influenced by the presence of bicarbonate. Consistent with our previously reported data, amantadine uptake could also be described by two transport sites, a high-affinity-capacity site that is bicarbonate-dependent and a lower-affinity-capacity transport site that is bicarbonate-independent. The renal tubule uptake of amantadine into proximal and distal tubules, in Krebs-Henseleit solution or Cross-Taggart buffers, was not inhibited by 10 to 1000 microM of TEA. However, tubule accumulation of TEA could be inhibited (>90%) by amantadine in proximal and distal tubules in Krebs Henseleit solution and Cross-Taggart buffers. In proximal tubules, N1 methylnicotinamide was not able to inhibit amantadine uptake but it reduced TEA uptake by 60 to 70% at similar concentrations. These data support the existence of multiple renal tubule organic cation transporters that have different substrate affinity and controlling mechanisms. It is also apparent that amantadine characterizes organic cation transporters that are distinct from those characterized by TEA. PMID- 10381791 TI - gamma-Hydroxybutyrate modulates synthesis and extracellular concentration of gamma-aminobutyric acid in discrete rat brain regions in vivo. AB - gamma-Hydroxybutyrate possesses most of the properties of a neurotransmitter/neuromodulator that acts via specific pathways and receptors in brain. Beside its regulatory effects on dopaminergic transmission, gamma hydroxybutyrate was thought for many years to interfere with gamma-aminobutyric acid (GABA)ergic processes in the brain. The present study demonstrates that in the rat frontal cortex in vivo, gamma-hydroxybutyrate or its agonist NCS-356 administered systemically at a high dose (500 mg/kg) increases GABA contents in dialysates via a mechanism blocked by the peripheral administration of the gamma hydroxybutyrate antagonist NCS-382. Under the same conditions, the extracellular concentration of this amino acid was not modified in the hippocampus. However, when administered at a low dose (250 mg/kg), gamma-hydroxybutyrate decreases GABA content of the dialysates of the frontal cortex by an NCS-382-sensitive mechanism. Spontaneous [3H]GABA release was observed in the frontal cortex of rats at 160 min after i.p. [3H]-gamma-hydroxybutyrate administration. This result indicates that gamma-hydroxybutyrate in vivo could be the precursor of an extracellular GABA pool in the frontal cortex. After i.p. [3H]-gamma hydroxybutyrate administration in the rat, the amino acid contents of several brain regions were quantified 160 min later, and the radioactivity in each region was measured. [3H]GABA, [3H]glutamate, and [3H]glycine were detected in most, but not all, of the brain regions studied. In particular, radioactive GABA was not detected in the hippocampus. The other amino acids were not labeled. These results show that gamma-hydroxybutyrate modulates the synthesis and the extracellular concentrations of GABA in specific regions of the rat brain. Identification of these GABA pools and determination of their functional role remain to be defined. PMID- 10381792 TI - L-Citrulline, the by-product of nitric oxide synthesis, decreases vascular smooth muscle cell proliferation. AB - Endothelium injury plays an important role in atherosclerosis. Damage to the endothelium results in vascular smooth muscle cell proliferation. Natriuretic peptides present a potent antimitogenic action, mediating their biological effects via the binding of guanylate cyclase-linked atrial natriuretic peptide (ANP) receptor and the production of cyclic GMP. In a previous study, we demonstrated that L-citrulline, the by-product of nitric oxide synthesis, could relax rabbit aortic rings by stimulating the guanylate cyclase-linked ANP receptor. In this work, we investigated the effect of L-citrulline on vascular smooth muscle cell proliferation. L-Citrulline (10(-8) M) significantly decreased rat aortic (A10 cell line) vascular smooth muscle proliferation. The percentage of inhibition exerted by L-citrulline on days 3, 5, and 7 of the proliferation curve was 20.0 +/- 0.5%, 37.5 +/- 8.3%, and 28. 5 +/- 7.2%, respectively. In addition, L-citrulline also inhibited serum-induced DNA synthesis, measured as 5 bromo-2'-deoxyuridine incorporation. 5-Bromo-2'-deoxyuridine incorporation into nuclei of vehicle-treated cells was 40.5 +/- 2.4%, whereas in L-citrulline treated cells the percentage decreased to 36.0 +/- 4.1%, 29.1 +/- 2.0% (P <.01, n = 4), 30.5 +/- 2.4% (P <.05, n = 4), and 23.1 +/- 0.5% (P <.001, n = 4) for 10( 10), 10(-9), 10(-8), and 10(-7) M, respectively. Zaprinast, a phosphodiesterase type V inhibitor, enhanced 5-bromo-2'-deoxyuridine incorporation in serum stimulated cells. Moreover, L-citrulline inhibition of serum-stimulated DNA synthesis was abolished by HS-142-1 (10(-5) M), an ANP receptor antagonist. In another group of experiments, L-citrulline was shown to increase intracellular cyclic GMP levels from 2.1 +/- 0.2 pmol of cGMP/mg protein to 4.1 +/- 0.1 for L citrulline (10(-8) M) (P <.001, n = 3). These findings suggest that L-citrulline decreases vascular smooth muscle cell proliferation in the A10 cell line by acting on DNA synthesis by mechanisms that involve the ANP receptor. PMID- 10381793 TI - Binding and hydrolysis of meperidine by human liver carboxylesterase hCE-1. AB - Human liver carboxylesterases catalyze the hydrolysis of apolar drug or xenobiotic esters into more soluble acid and alcohol products for elimination. Two carboxylesterases, hCE-1 and hCE-2, have been purified and characterized with respect to their role in cocaine and heroin hydrolysis. The binding of meperidine (Demerol) and propoxyphene (Darvon) was examined in a competitive binding, spectrophotometric assay. The hCE-1 and hCE-2 bound both drugs, with Ki values in the 0.4- to 1.3-mM range. Meperidine was hydrolyzed to meperidinic acid and ethanol by hCE-1 but not hCE-2. The Km of hCE-1 for meperidine was 1.9 mM and the kcat (catalytic rate constant) was 0.67 min-1. Hydrolysis of meperidine by hCE-1 was consistent with its specificity for hydrolysis of esters containing simple aliphatic alcohol substituents. Hence, hCE-1 in human liver microsomes may play an important role in meperidine elimination. Propoxyphene was not hydrolyzed by hCE-1 or hCE-2. This observation is consistent with the absence of a major hydrolytic pathway for propoxyphene metabolism in humans. PMID- 10381794 TI - Postnatal ontogeny and hormonal regulation of sulfotransferase SULT1B1 in male and female rats. AB - The ontogenic and hormonal regulation of a sulfotransferase, SULT1B1, was examined. Hepatic RNA was isolated from rats of various ages from 1 to 90 days. The mRNA for SULT1B1 is low for both sexes until a dramatic increase ( approximately 6-fold) occurs between 15 and 30 days of age in male rats. SULT1B1 expression then decreases to half of the maximal level by 90 days of age. The increase in SULT1B1 mRNA in female rats is less dramatic and occurs between 30 and 45 days of age. SULT1B1 mRNA expression plateaus from 45 to 90 days in female rats. Expression of SULT1B1 mRNA is comparable in adult male and female rats. RNA was isolated from hypophysectomized (HX) animals and HX animals treated with growth hormone [by either male (injection) or female (infusion) pattern], estradiol, progesterone, or testosterone. HX and HX plus growth hormone, or HX plus steroid replacement, did not alter SULT1B1 mRNA expression. Pituitary-intact rats were treated with steroidal compounds dexamethasone (DEX) and pregnenolone 16alpha-carbonitrile (PCN). Both DEX and PCN increased expression of SULT1B1 mRNA in male rats (4- and 3-fold, respectively). However, in female rats, only PCN induced SULT1B1 mRNA (2-fold), whereas DEX did not induce SULT1B1 in female rats. Analysis of SULT1B1 protein expression indicated that only when SULT1B1 mRNA was markedly increased, that is in DEX-treated male rats, was SULT1B1 protein increased. Thus, although adult male and female rats have similar SULT1B1 mRNA expressions, the patterns develop ontogenically differently. SULT1B1 is not regulated by pituitary hormones and DEX induces SULT1B1 protein in male rats. PMID- 10381795 TI - Pharmacological reduction of small conductance calcium-activated potassium current (SK) potentiates the excitatory effect of ethanol on ventral tegmental area dopamine neurons. AB - Dopaminergic neurons in the ventral tegmental area (VTA) are important for the rewarding properties of drugs of abuse, including ethanol. We previously demonstrated that ethanol excites VTA neurons and that ethanol reduces the amplitude of the after hyperpolarization (AHP) that follows spontaneous action potentials. Because the small conductance calcium-activated potassium current (SK) is a component of the AHP of VTA neurons, we assessed the effect of the SK blockers apamin and d-tubocurarine (d-TC) on the action of ethanol on dopaminergic VTA neurons with intracellular and extracellular recording in rat brain slices. Apamin (1-200 nM) and d-TC (100 and 400 microM) caused concentration-dependent reductions in the AHP amplitude. Ethanol (80 mM) caused a small reduction in the AHP. In the presence of apamin (40 nM), ethanol (80 mM) caused a much larger reduction in AHP amplitude. Extracellular studies showed that apamin (20, 40, and 100 nM) and d-TC (400 microM) had no significant effect on the spontaneous firing rate of dopaminergic VTA neurons but enhanced the potency of ethanol to increase their firing rate. These results indicate that the ethanol-induced reduction of the AHP and excitation of VTA neurons is not due to a reduction in SK current. Furthermore, blockade of SK current by apamin or d-TC enhances the excitatory effect of ethanol on dopaminergic VTA neurons. These data suggest that the amount of SK current present affects the sensitivity of dopaminergic VTA neurons to ethanol excitation and that neurotransmitters that reduce SK current may increase the reward potency of ethanol. PMID- 10381796 TI - Characterization of 2-[125I]iodomelatonin binding sites in Syrian hamster peripheral organs. AB - The neurohormone melatonin is a key agent in synchronizing the circadian rhythms. At least three types of binding sites have been described for melatonin: the G coupled, seven-transmembrane domain receptors mt1 and MT2 and a putative binding site called MT3. The latter has been described in hamster brain membranes, and its binding capacity is optimum at 4 degrees C. We further characterized this binding site on other peripheral hamster tissues, including intestine, liver, kidney, lung, muscle, and heart. We found a high level of binding sites (>30 fmol/mg of protein) in intestine and kidney. Furthermore, we completed the existing pharmacological profile of this site, which can now be described as 2 iodomelatonin > 6-chloromelatonin > methy-isobutyl-amiloride > acridine orange > 5-methylcarbonylamino-N-acetyltryptamine > prazosin > N-acetylserotonin > melatonin. This profile was found in all the hamster organs tested that had a large number of binding sites, namely, brain, intestine, kidney and liver. Furthermore, when comparisons were possible, the MT3 pharmacological characteristics were similar to those described in the literature for hamster brain and testis. This profile was compared to the pharmacology obtained on human cloned mt1 and MT2 receptors and proved to be completely different, as expected. We provide new evidence for an alternate melatonin binding site not only in hamster brain but also in some peripheral organs. PMID- 10381797 TI - Stimulation of epithelial sodium channel activity by the sulfonylurea glibenclamide. AB - The amiloride-sensitive epithelial sodium channel (ENaC) contributes to the regulation of the sodium balance and blood pressure because it mediates a rate limiting step in sodium transport across the epithelium of the distal nephron. The activity of ENaC is regulated by hormones, such as aldosterone and vasopressin, and by other intracellular or extracellular factors, but the mechanisms of these regulations are not yet well understood. It has been proposed that ENaC may be regulated by an associated ATP-binding cassette protein such as the cystic fibrosis conductance regulator or the K channel-associated sulfonylurea receptor. Glibenclamide, a known inhibitor of sulfonylurea receptor and cystic fibrosis conductance regulator, induced a dose-dependent and reversible stimulation (of the order of 40-50%) of the amiloride-sensitive current in oocytes expressing Xenopus ENaC, with a K1/2 of 45 +/- 5 microM. A similar effect was observed in oocytes expressing human ENaC, but not rat ENaC. Measurements performed with various combinations of rat and Xenopus subunits indicated that several subunits are involved in this effect. Glibenclamide also increased the transepithelial Na transport by the A6 Xenopus kidney cell line. Single-channel current recordings showed a doubling of the number of the open channels when glibenclamide was applied locally to the extracellular surface of the cell membrane. These results support the hypothesis of the existence of an associated ATP-binding cassette-type regulatory protein associated with the epithelial sodium channel. PMID- 10381798 TI - Neuroprotection by a novel compound, NS521. AB - NS521 (1-(1-butyl)-4-(2-oxo-1-benzimidazolinyl)piperidine) belongs to a group of novel benzimidazolones, which exhibit neurotrophic-like activities. In vitro, NS521 rescued neuronal PC12 cells from death induced by serum and nerve growth factor deprivation. The survival effect of NS521 appeared to reflect a delay of the apoptotic process, because the extent of DNA fragmentation was attenuated transiently by NS521. NS521 did not preserve the neurites of the rescued cells, which, otherwise, appeared to be healthy and were able to regenerate when serum and nerve growth factor were added back to the culture. In vivo, NS521 provided significant protection against the delayed loss of hippocampal CA1 neurons in a gerbil model of transient global ischemia. A neuroprotective effect of NS521 in the peripheral nervous system also was observed in rats after transection of the sciatic nerve, where daily treatment with NS521 was found to inhibit retrograde degeneration of the transected nerve. The neuroprotective effect of NS521 is unlikely to be mediated through neurotrophin receptors, such as TrkA, because NS521 did not induce phosphorylation of the 44- and 42-kDa isoforms of mitogen activated protein kinases (ERK1/2) in PC12 cells. PMID- 10381799 TI - Peripheral effects of the kappa-opioid agonist EMD 61753 on pain and inflammation in rats and humans. AB - The objective of the present study was to evaluate the effects of EMD 61753 (asimadoline), a kappa-opioid receptor agonist with restricted access to the central nervous system, on postoperative pain in patients who underwent knee surgery and on nociceptive thresholds and inflammation in rats treated with Freund's complete adjuvant. Patients treated with EMD 61753 (10 mg p.o.) tended to report an increase in pain, as evaluated by a visual analog scale and by the time to the first request for and the total amount of supplemental analgesic medication. The global tolerability of EMD 61753 was assessed as significantly inferior to that of a placebo by the investigator. In rats, the bilateral intraplantar (i.pl.) injection of EMD 61753 (0.1-3.2 mg) resulted in dose dependent antinociception in both inflamed and noninflamed paws, with a peak at 5 min after injection, as evaluated by the paw pressure method. However, at later time points (1 h-4 days), a significant decrease in the paw pressure threshold was observed, confirming its tendency toward a hyperalgesic action in humans. This was accompanied by an increase in paw volume and paw temperature, with a peak at 6 h after injection. EMD 61753 (1.6 mg)-induced analgesia was blocked by the peripheral opioid receptor antagonist naloxone methiodide (2.5-10 mg/kg s.c.) and by the kappa receptor antagonist nor-binaltorphimine (0.1 mg; i.pl.). In contrast, EMD 61753 (1.6 mg)-induced hyperalgesia and increases in paw volume and paw temperature were blocked neither by naloxone methiodide (10-40 mg/kg s.c.) nor by dizocilpine maleate (0.003-0.009 mg i.pl.), a N-methyl-D-aspartic acid receptor antagonist. These data show differentially mediated peripheral actions of EMD 61753: kappa-opioid receptor-induced analgesia and nonopioid, non-N-methyl D-aspartic acid hyperalgesic and proinflammatory effects. PMID- 10381800 TI - Ethanol directly depresses AMPA and NMDA glutamate currents in spinal cord motor neurons independent of actions on GABAA or glycine receptors. AB - Ethanol is a general anesthetic agent as defined by abolition of movement in response to noxious stimulation. This anesthetic endpoint is due to spinal anesthetic actions. This study was designed to test the hypothesis that ethanol acts directly on motor neurons to inhibit excitatory synaptic transmission at glutamate receptors. Whole cell recordings were made in visually identified motor neurons in spinal cord slices from 14- to 23-day-old rats. Currents were evoked by stimulating a dorsal root fragment or by brief pulses of glutamate. Ethanol at general anesthetic concentrations (50-200 mM) depressed both responses. Ethanol also depressed glutamate-evoked responses in the presence of tetrodotoxin (300 nM), showing that its actions are postsynaptic. Block of inhibitory gamma aminobutyric acidA and glycine receptors by bicuculline (50 microM) and strychnine (5 microM), respectively, did not significantly reduce the effects of ethanol on glutamate currents. Ethanol also depressed glutamate-evoked currents when the inhibitory receptors were blocked and either D, L-2-amino-5 phosphonopentanoic acid (40 microM) or 6-cyano-7-nitroquinoxaline-2,3-dione disodium (10 microM) were applied to block N-methyl-D-aspartate or alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid/kainate receptors, respectively. The results show that ethanol exerts direct depressant effects on both alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid and N-methyl-D-aspartate glutamate currents in motor neurons. Enhancement of gamma-aminobutyric acidA and glycine inhibition is not required for this effect. Direct depression of glutamatergic excitatory transmission by a postsynaptic action on motor neurons thus may contribute to general anesthesia as defined by immobility in response to a noxious stimulus. PMID- 10381801 TI - Enhanced antitumor potency of polyethylene glycolylated tumor necrosis factor alpha: a novel polymer-conjugation technique with a reversible amino-protective reagent. AB - We attempted to develop a novel method for the chemical modification of cytokines with synthetic polymers to increase in vivo therapeutic efficacy. A pH-reversible amino-protective reagent, dimethylmaleic anhydride (DMMAn), was used for polymer conjugation of tumor necrosis factor-alpha (TNF-alpha) with polyethylene glycol (PEG). The novel PEGylated TNF-alpha, PEG-TNF-alpha(+), which was pretreated with DMMAn before PEGylation, had 20% to 40% higher specific activity than PEG-TNF alpha(-) (not treated with DMMAn) in vitro. Moreover, PEG-TNF-alpha(+) more potently caused tumor necrosis in Meth-A solid tumors in mice than did PEG-TNF alpha(-). The middle fraction (M) of PEG-TNF-alpha(+), which was of the optimal degree of modification among PEG-TNF-alpha(+)s with different molecular weights, caused the highest degree of tumor hemorrhagic necrosis: 30-fold higher than native TNF-alpha and 2-fold higher than the most potent MPEG-TNF-alpha(-) that also had nearly the same molecular weight. Significantly, improvements in antitumor activity in vivo were more marked than were changes in specific activity. Furthermore, native TNF-alpha caused a dose-dependent body weight loss in mice, whereas no obvious side effects were observed in any PEG-TNF-alpha treated mice. These results suggest that PEGylation using DMMAn is a useful for clinical cytokine delivery. PMID- 10381802 TI - Efficient extraction by the liver governs overall elimination of hepatocyte growth factor in rats. AB - A steady-state pharmacokinetic analysis was performed to investigate the overall elimination and extraction of hepatocyte growth factor (HGF) by its target organs, including liver, kidney, and lung, during its constant i.v. infusion in rats. The plasma clearance of HGF became saturated as the steady-state plasma concentration (Cpss) increased, but complete saturation was not achieved, even when the Cpss ( approximately 1000 pM) was much higher than the dissociation constant for the HGF receptor (20-40 pM), which has been identified as one of the major clearance sites for HGF. This result suggests that there is a low-affinity and high-capacity clearance mechanism, other than receptor-mediated endocytosis, involved in its elimination from the body. The hepatic extraction ratio of HGF, assessed by determining the HGF concentration in both the circulating blood and hepatic vein, was 40 to 60%, whereas the HGF extraction both in kidney and lung was always less than 10%. Hepatic clearance accounted for approximately 70% of the plasma clearance at any Cpss. Thus, the present study shows that HGF in circulating plasma is efficiently extracted by the liver compared with other HGF target organs, the liver being involved in 70% of the overall elimination both under linear and nonlinear conditions. Biliary excretion of HGF was observed, but this accounted for only 0.1 to 0. 2% of the infusion rate, indicating that the nonlysosomal pathway of HGF, which avoids the lysosomal enzymes and transcytoses HGF directly into the bile, is very minor indeed. PMID- 10381803 TI - Long-lasting changes of rat blood pressure to vasoconstrictors and vasodilators induced by nitric oxide donor infusion: involvement of potassium channels. AB - We investigated the effects of the exposure of the rat vascular system to nitric oxide (NO), using infusion of either NO donor sodium nitroprusside (SNP) or S nitroso-acetyl-DL-penicillamine (SNAP) on mean arterial pressure (MAP) responses to vasoconstrictors (phenylephrine, angiotensins I and II) and to vasodilators (bradykinin, acetylcholine, SNP, and iloprost). SNP (250 nmol/kg/ min) or SNAP (85 nmol/kg/min) infused for 30 min decreased MAP by 40 to 60 mm Hg. MAP returned to normal levels 5 to 10 min after the end of infusion. After infusion of SNP or SNAP the effects of phenylephrine, angiotensin I, and angiotensin II were reduced by 40 to 80%, whereas the responses to bradykinin or acetylcholine were enhanced by 50 to 80%. These changes in vascular responsiveness persisted for at least 24 h after the SNP infusion. Pretreatment with either tetraethylammonium (360 micromol/kg) or 4-aminopyridine (4-AP; 1 micromol/kg) did not alter the effects of phenylephrine or bradykinin in control animals, but prevented SNP-induced changes in responsiveness to phenylephrine or bradykinin. On the other hand, administration of tetraethylammonium, even 24 h after SNP infusion, reversed hyporesponsiveness to phenylephrine, whereas 4-AP was ineffective. Tetraethylammonium and 4-AP did not alter the increased responses to bradykinin. Glibenclamide was without effect in any situation. These results indicate that NO induced changes on vascular responsiveness to vasoconstrictors and vasodilators are much more profound and long-lasting than described previously and that the effects of NO appear to be, at least in part, mediated by persistent activation of a tetraethylammonium-sensitive population of K+ channels. PMID- 10381804 TI - Transport characteristics of diphenhydramine in human intestinal epithelial Caco 2 cells: contribution of pH-dependent transport system. AB - Transport characteristics of diphenhydramine, an antihistamine, were studied in cultured human intestinal Caco-2 cell monolayers to elucidate the mechanisms of its intestinal absorption. Diphenhydramine accumulation in the monolayers increased rapidly and was influenced by extracellular pH (pH 7.4 > 6.5 > 5.5). Diphenhydramine uptake was temperature dependent, saturable, and not potential sensitive. Kinetic analysis revealed that the apparent Km values were constant (0.8-1.0 mM) in all pH conditions tested, whereas Vmax values decreased at the lower pH. The initial uptake of diphenhydramine was competitively inhibited by another antihistamine, chlorpheniramine, with a Ki value of 1.3 mM. On the other hand, cimetidine and tetraethylammonium, typical substrates for the renal organic cation transport system, had no effect. Moreover, biological amines and neurotransmitters, such as histamine, dopamine, serotonin, and choline, also had no effect on the diphenhydramine accumulation. Finally, diphenhydramine uptake was stimulated by preloading monolayers with chlorpheniramine (trans-stimulation effect). These findings indicate that diphenhydramine transport in Caco-2 cells is mediated by a specific transport system. This pH-dependent transport system may contribute to the intestinal absorption of diphenhydramine. PMID- 10381805 TI - Effects of response contingent and noncontingent cocaine injection on hypothalamic-pituitary-adrenal activity in rhesus monkeys. AB - Earlier studies of cocaine's effects on the hypothalamic-pituitary-adrenal (HPA) axis used nonresponse-contingent designs in which the investigator determined dose, timing, and route of administration. It is important to evaluate whether "control" over cocaine delivery is a significant determinant of cocaine's HPA axis effect. This study measured cocaine's effects on plasma adrenocorticotropic hormone and cortisol, using nonresponse-contingent injections followed later by response-contingent cocaine delivery. In addition, the effects of cocaine history on the HPA response to a noncontingent injection of 1 mg/kg of cocaine were measured. HPA effects of corticotropin-releasing hormone (CRF) were also measured. Male and female rhesus monkeys, with surgically placed venous catheters, were tested in their home cages. Up to 13 injections of saline and cocaine (0.01-, 0.03-, 0.1-, and 0.3-mg/kg/injection) were administered at 10-min intervals (nonresponse-contingent condition) and on a fixed ratio 30, time out 10 min schedule of reinforcement. Overall, cocaine delivered response contingently produced larger, more dose-dependent HPA responses than did noncontingent delivery. The HPA response to a 1 mg/kg cocaine infusion in cocaine-naive monkeys was not predictive of the HPA effect of this dose subsequent to acquisition of cocaine self-administration. Overall, male monkeys had larger HPA responses to cocaine than did female monkeys. Finally, the HPA effects of CRF were significantly correlated with those of large cocaine doses delivered nonresponse contingently, but not with response-contingent administration. PMID- 10381807 TI - Impact of ethnic origin and quinidine coadministration on codeine's disposition and pharmacodynamic effects. AB - CYP2D6 is polymorphically distributed so that in poor metabolizers enzyme activity is missing. The goal of this study was to compare the pharmacokinetics and pharmacodynamics of codeine with and without quinidine between Caucasian and Chinese extensive metabolizers of debrisoquin. Nine Caucasians and eight Chinese subjects received in random, double blind fashion, on two occasions, codeine 120 mg. with placebo or with quinidine 100 mg. Pharmacodynamic effects were determined over 6 h. Codeine-apparent clearance and partial metabolic clearance by O-demethylation were significantly greater in the Caucasian than in the Chinese subjects (1939 +/- 175 ml/min versus 1301 +/- 193 ml/min, p <.03 and 162.7 +/- 36.6 ml/min versus 52.7 +/- 12.7 ml/min, p <.02, respectively). Codeine's respiratory effects (except on resting ventilation) were significantly greater in the Caucasian than in the Chinese subjects (p <.05), but no interethnic differences were noted in codeine's effect on the digit symbol substitution test and pupillary ratio. No morphine or morphine metabolites were detected in plasma when codeine was coadministered with quinidine. Codeine O demethylation was significantly reduced after quinidine in both ethnic groups; however, the absolute decrease was greater in Caucasians (115.8 +/- 25.9 ml/min versus 46.8 +/- 10.6 ml/min, respectively, p <.03). The diminished production of morphine after quinidine was associated in the Caucasians, but not in the Chinese, with a marked reduction in codeine's effects (p <.01). In conclusion, Chinese produce less morphine from codeine, exhibit reduced sensitivity to that morphine, and therefore might experience reduced analgesic effect in response to codeine. In addition, quinidine induced inhibition of codeine O-demethylation is ethnically dependent with the reduction being greater in Caucasians. PMID- 10381806 TI - Moxonidine, a selective alpha2-adrenergic and imidazoline receptor agonist, produces spinal antinociception in mice. AB - alpha2-Adrenergic receptor (AR)-selective compounds produce antihypertensive and antinociceptive effects. Moxonidine alleviates hypertension in multiple species, including humans. This study demonstrates that intrathecally administered moxonidine produces antinociception in mice. Antinociception was detected via the (52.5 degrees C) tail-flick and Substance P (SP) nociceptive tests. Moxonidine was intrathecally administered to ICR, mixed C57BL/6 x 129/Sv [wild type (WT)], or C57BL/6 x 129/Sv mice with dysfunctional alpha2aARs (D79N-alpha2a). The alpha2AR-selective antagonist SK&F 86466 and the mixed I1/alpha2AR-selective antagonist efaroxan were tested for inhibition of moxonidine-induced antinociception. Moxonidine prolonged tail-flick latencies in ICR (ED50 = 0.5 nmol; 0. 3-0.7), WT (0.17 nmol; 0.09-0.32), and D79N-alpha2a (0.32 nmol; 0. 074 1.6) mice. Moxonidine inhibited SP-elicited behavior in ICR (0. 04 nmol; 0.03 0.07), WT (0.4 nmol; 0.3-0.5), and D79N-alpha2a (1.1 nmol; 0.7-1.7) mice. Clonidine produced antinociception in WT but not D79N-alpha2a mice. SK&F 86466 and efaroxan both antagonized moxonidine-induced inhibition of SP-elicited behavior in all mouse lines. SK&F 86466 antagonism of moxonidine-induced antinociception implicates the participation of alpha2ARs. The comparable moxonidine potency between D79N-alpha2a and WT mice suggests that receptors other than alpha2a mediate moxonidine-induced antinociception. Conversely, absence of clonidine efficacy in D79N-alpha2a mice implies that alpha2aAR activation enables clonidine-induced antinociception. When clinically administered, moxonidine induces fewer side effects relative to clonidine; moxonidine-induced antinociception appears to involve a different alpha2AR subtype than clonidine induced antinociception. Therefore, moxonidine may prove to be an effective treatment for pain with an improved side effect profile. PMID- 10381808 TI - Activity of putative cognition enhancers in kynurenate test performed with human neocortex slices. AB - Some cognition enhancers were previously shown to potently prevent antagonism of the N-methyl-D-aspartate (NMDA)-evoked release of norepinephrine (NE) brought about in slices of rat hippocampus by kynurenic acid, an endogenous NMDA receptor blocker. We have examined the impact of putative nootropic agents in the kynurenate test performed with slices of human cerebral cortex from patients undergoing neurosurgery. In slices of human neocortex, local application of NMDA evoked release of [3H]NE; the effect of NMDA was antagonized by several NMDA receptor antagonists, including kynurenic acid. The antagonism of the NMDA-evoked [3H]NE release produced by 300 microM kynurenate was potently (EC50 <10 microM) prevented by most of the nootropics tested, including aniracetam, oxiracetam, D cycloserine, and the glutamate analog CR 2249 (but not its enantiomer CR 2361). Nicotine or tacrine (up to 10 microM) did not show any effect in the kynurenate test. Nicotine (30-100 microM) itself increased the release of [3H]NE; interestingly, the nicotine-evoked overflow was blocked not only by the nicotin receptor antagonist mecamylamine but also by NMDA receptor antagonists, suggesting an indirect mechanism mediated by glutamate/aspartate release. To conclude, the similarities between the data obtained here with human neocortex slices and those previously obtained in the rat indicate that the kynurenate test performed with rat brain slices may represent a useful biochemical assay to study cognition-enhancing drugs. PMID- 10381809 TI - Three- and four-dimensional quantitative structure activity relationship analyses of cytochrome P-450 3A4 inhibitors. AB - The program Catalyst was used to build three-dimensional quantitative structure activity relationship (3D-QSAR) pharmacophore models of the structural features common to competitive-type inhibitors of cytochrome P-450 (CYP) 3A4. These were compared with 3D- and four-dimensional (4D)-QSAR partial least-squares (PLS) models built using molecular surface-weighted holistic invariant molecular (MS WHIM) descriptors for size and shape of the inhibitor. The Catalyst pharmacophore model generated from multiple conformers of competitive inhibitors of CYP3A4 mediated midazolam 1'-hydroxylation (n = 14) yielded a high correlation of observed and predicted Ki values of r = 0.91. Similarly, PLS MS-WHIM was used to produce 3D- and 4D-QSARs for this data set and produced models that were statistically predictable after cross-validation. Two additional Catalyst pharmacophores were constructed from literature Ki values (n = 32) derived from the inhibition of CYP3A-mediated cyclosporin A metabolism and IC50 data (n = 22) from the inhibition of CYP3A4-mediated quinine 3-hydroxylation. These Catalyst pharmacophores illustrated correlations of observed and predicted inhibition for CYP3A4 of r = 0.77 and 0.92, respectively. The corresponding 4D-QSARs generated by PLS MS-WHIM for these data sets were of comparable quality as judged by cross validation. Both Ki pharmacophores generated with Catalyst were also validated by predicting the Ki(apparent) values of a test set of eight CYP3A4 inhibitors not included in either model. In seven of eight cases, the residuals of the predicted Ki(apparent) values were within 1 log unit of the observed values. The 3D- and 4D QSAR models produced in this study suggest the utility of future in silico prediction of CYP3A4-mediated drug-drug interactions. PMID- 10381810 TI - Brevetoxins cause acute excitotoxicity in primary cultures of rat cerebellar granule neurons. AB - Brevetoxins (designated PbTx-1 to -10) are potent lipid-soluble polyether compounds that are known to bind to and modulate voltage-gated sodium channel activity. To investigate whether brevetoxins produce direct central nervous system neurotoxic effects, cultured rat cerebellar granule neurons were exposed to brevetoxins in Locke's buffer for 2 h at 22 degrees C. Neuronal injury was quantified by assaying lactate dehydrogenase activity in the exposure buffer and in conditioned growth media collected at 22 h after brevetoxin exposure. Brevetoxins produced acute neuronal injury and death in neurons with a rank order potency of PbTx-1 (EC50 = 9.31 +/- 0.45 nM) > PbTx-3 (EC50 = 53.9 +/- 2.8 nM) > PbTx-2 (EC50 = 80.5 +/- 5.9 nM) > PbTx-6 (EC50 = 1417 +/- 32 nM), which is similar to their previously determined rank order potency for brevetoxin-induced icthyotoxicity and binding to [3H]PbTx-3-labeled sodium channels on synaptosomes. The neurotoxic response could be prevented by coapplication of the sodium channel antagonist tetrodotoxin or by the competitive or noncompetitive N-methyl-D aspartate (NMDA) receptor antagonists D-AP5 and MK-801, ketamine, dextromethorphan, and dextrorphan, respectively. NMDA receptor antagonists afforded neuroprotection with rank order potencies comparable to those measured previously for protection against glutamate-induced excitotoxic responses. Further analysis revealed that brevetoxins induced a concentration-dependent release of L-glutamate and L-aspartate into the exposure buffer. These data indicate that brevetoxin-induced injury in cultured rat cerebellar granule neurons is mediated by NMDA receptors that are activated indirectly as a consequence of PbTx-induced sodium channel activation and attendant excitatory amino acid release. PMID- 10381811 TI - Identification of selective mechanism-based inactivators of cytochromes P-450 2B4 and 2B5, and determination of the molecular basis for differential susceptibility. AB - Rabbit cytochromes P-450 (P-450) 2B4 and 2B5 differ by only 12 amino acid residues yet they exhibit unique steroid hydroxylation profiles. Previous studies have led to the identification of active site residues that are determinants of these specificities. In this study, mechanism-based inactivators were identified that discriminate between the closely related 2B4 and 2B5 enzymes. A previously characterized inhibitor, 2-ethynylnaphthalene (2EN), was found to be selective for 2B4 inactivation. As inhibitor metabolism and the partition ratio affect susceptibility, molecular dynamics simulations were performed to assess the stability of the productive binding orientation of 2EN within 2B4 and 2B5 three dimensional models. Although 2EN was stable within the 2B4 model, it exhibited substantial movement away from the heme moiety in the 2B5 model. However, heterologously expressed 2B5 was found to catalyze the oxidation of 2EN to the stable product 2-naphthylacetic acid. Thus, the increased mobility of 2EN may result in reduced susceptibility of 2B5 by increasing the probability that the reactive ketene intermediate hydrolyzes with water instead of reacting with active site residues. Another compound, 1-adamantyl propargyl ether (1APE), selectively inactivated 2B5. The structural basis for 2EN and 1APE susceptibility was assessed using active site mutants. Interconversion of 2EN susceptibility was observed for 2B4 or 2B5 mutants containing a single alteration at residue 363. Single substitutions in 2B4 also conferred susceptibility to 1APE; however, multiple alterations were required to reduce the susceptibility of 2B5. These alterations may influence inhibitor susceptibility by affecting the stability of the productive binding orientation. PMID- 10381812 TI - Expression of multiple alpha1-adrenoceptors on vascular smooth muscle: correlation with the regulation of contraction. AB - Previous work has shown that the genes encoding each alpha1-adrenoceptor subtype are coexpressed throughout the peripheral vascular system. We have evaluated subtype-selective antibodies as tools to determine the extent of protein expression in arteries. The alpha1A-, alpha1B-, and alpha1D-adrenoceptors were detected in the medial layer of the aorta, caudal, femoral, iliac, renal, superior mesenteric, and mesenteric resistance arteries. In Rat1 fibroblasts expressing each subtype, immunoreactivity was noted both on the cell surface and in a perinuclear orientation. Intense alpha1B-adrenoceptor immunostaining was similarly localized in cultured femoral and renal vascular smooth muscle cells. Although the cellular localization appeared to be the same, immunoreactivity obtained with alpha1A- and alpha1D-adrenoceptors was much less intense than that with the alpha1B-adrenoceptor. The alpha1A-adrenoceptor selective agonist A-61603 was 22-fold more potent in activating renal artery contraction when compared with the femoral artery. The expression of each alpha1-adrenoceptor was significantly decreased by in vivo application of antisense oligonucleotides targeted against each subtype. Inhibition of the expression of only one, the alpha1A in renal and the alpha1D in femoral arteries, reduced the contractile response to naphazoline. The results show: 1) subtype-selective antibodies can be used in tissues and cell culture to localize the alpha1-adrenoceptor subtypes, 2) in addition to expression on the cell surface, the alpha1-adrenoceptors are expressed intracellularly, and 3) despite expression of all adrenoceptors, a single subtype mediates the contractile response in the femoral and renal arteries. PMID- 10381813 TI - Efficacy of keratinocyte growth factor-2 in dextran sulfate sodium-induced murine colitis. AB - The purpose of this study was to determine the efficacy of a novel human protein, keratinocyte growth factor-2 (KGF-2), in a model of murine colitis induced by ad libitum exposure to a 4% solution of dextran sulfate sodium (DSS) in the drinking water. Initial evaluation of KGF-2 was based on its ability to reduce weight loss, stool score, and histological score in mice exposed to DSS for 7 days. When KGF-2 (0.1-10.0 mg/kg i.p. or s.c.) was injected daily into DSS-treated mice from day 0 to 7, it significantly reduced all three parameters in a dose-response fashion, with a minimum effective dose of between 1 and 3 mg/kg. When KGF-2 was given therapeutically, starting 4 days after initiation of the 7-day DSS treatment, the 3- but not the 0.5-mg/kg dose significantly enhanced weight recovery after discontinuation of DSS treatment. When DSS treatment was prolonged beyond the normal 7 days, therapeutic intervention on day 2 or 4 also significantly reduced mortality, weight loss, and stool score at the 1- and 3 mg/kg dose. Therapeutic treatment also resulted in reduction of colon myloperoxidase levels by more than 50%. These experiments suggest that KGF-2 may be clinically useful in the treatment of inflammatory bowel diseases such as ulcerative colitis and Crohn's disease. PMID- 10381814 TI - Hypothesis: sex determination and germline formation are committed at the pronucleate stage in mammalian embryos. PMID- 10381815 TI - Human granulosa cells express integrin alpha2 and collagen type IV: possible involvement of collagen type IV in granulosa cell luteinization. AB - Previously, it has been shown that integrin alpha6beta1 expressed on human granulosa cells regulates luteinization in co-operation with its ligand, laminin. In this study, integrin alpha2 was immunohistochemically demonstrated to be expressed on granulosa and large luteal cells. It was also detected on luteinizing theca interna cells after ovulation. Immunoreactive collagen type IV, which is one of the ligands for integrin alpha2beta1, was detected around granulosa cells in the pre-ovulatory follicles and its expression was rapidly increased during ovulation. By flow cytometry, collagen type IV was detected on the cell surface of luteinizing granulosa cells isolated from pre-ovulatory follicles, confirming the physiological interaction between granulosa cells and collagen type IV. Collagen type IV in follicular fluid was positively related with progesterone concentration. In 4-day cultures of granulosa cells, collagen type IV in the media was significantly increased by human chorionic gonadotrophin (HCG). The progesterone production was significantly attenuated when granulosa cells were cultured on collagen type IV-coated dishes, suggesting that collagen type IV suppresses granulosa cell luteinization. These findings show that collagen type IV, a ligand for integrin alpha2beta1, is rapidly produced around luteinizing granulosa cells during ovulation, probably under the control of luteinizing hormone (LH) and suggest that collagen type IV is a new parameter and/or regulator of granulosa cell luteinization in the periovulatory phases. PMID- 10381816 TI - Pentoxifylline-stimulated capacitation and acrosome reaction in hamster spermatozoa: involvement of intracellular signalling molecules. AB - We investigated the role of cAMP/cGMP, protein kinases and intracellular calcium ( [Ca2+]i) in pentoxifylline-stimulated hamster sperm capacitation and the acrosome reaction (AR) in vitro. Treatment with pentoxifylline (0.45 mM) initially increased sperm cAMP values 2.8-fold, compared with untreated controls (396 +/- 9.2 versus 141 +/- 6.0 fmoles/10(6) spermatozoa; mean +/- SEM, n = 6) after 15 min, although by 3 h, cAMP values were similar (503-531 fmoles/10(6) spermatozoa). cGMP values ( approximately 27 fmoles/10(6) spermatozoa) were the same in treated and control spermatozoa. Both sperm capacitation and the AR, determined from the absence of an acrosomal cap, were stimulated by pentoxifylline; these were almost completely inhibited by a Cl-/ HCO3- antiporter inhibitor (4,4-diisothiocyanato-stilbene-2,2 disulphonic acid; 1 mM) defined from the degree of sperm motility and by a protein kinase A inhibitor (H89; 10 microM). A protein kinase C inhibitor (staurosporine, 1 nM) did not affect pentoxifylline-stimulated capacitation but inhibited the AR by 50%. A protein tyrosine kinase inhibitor (tyrphostin A-47, 0.1 mM) had no effect on either pentoxifylline-stimulated capacitation or AR. A phospholipase A2 inhibitor (aristolochic acid, 0.4 mM) markedly inhibited the pentoxifylline-stimulated AR but not capacitation. When intracellular sperm calcium [Ca2+/-]i was measured using fura-2-AM, there was an early rise (271 nM at 0.5 h) in pentoxifylline treated spermatozoa; this appeared to be due to intracellular mobilization rather than to uptake. In the absence of extracellular Ca2+, sperm motility was maintained in the presence of pentoxifylline, but capacitation did not occur; spermatozoa exhibited a low level of hyperactivated motility and had a poor rate of AR (20.5 +/- 2.3%). These results suggest that: (i) the pentoxifylline stimulated early onset of sperm capacitation may be mediated by an early rise in cAMP and [Ca2+/-]i and involves protein kinase A activity; and (ii) pentoxifylline-stimulated AR may require phospholipase A2 and protein kinase C activity. PMID- 10381817 TI - Outer dense fibre proteins from human sperm tail: molecular cloning and expression analyses of two cDNA transcripts encoding proteins of approximately 70 kDa. AB - The outer dense fibres (ODF) are a main cytoskeletal structure of the sperm tail. Despite their importance in the morphology and function of the sperm tail, their constituents are poorly described. Here we investigate the protein composition of human outer dense fibres. Our results suggest that human ODF consist of about 10 major and of at least 15 minor proteins, where all major proteins are ODF1, ODF2 or ODF2-related proteins. From a human testis cDNA library, we isolated two slightly different cDNAs encoding ODF2 proteins of approximately 70 kDa. Human ODF2 cDNAs and their encoded proteins are very similar to those isolated from rat and mouse pointing to a high evolutionary pressure residing on these proteins. Transcription of ODF2 is restricted to testis tissue and more specifically to round spermatids as was demonstrated by a non-radioactive in-situ hybridization. ODF2 proteins were detected in the sperm tail. Their distribution along the length of the sperm tail shows that the ODF normally extend to about half the principal piece of the sperm tail. The former result opens the possibility for a screening regarding the distribution of sperm tail proteins related to motility disorders. PMID- 10381819 TI - Macrophage derived growth factors modulate Fas ligand expression in cultured endometrial stromal cells: a role in endometriosis. AB - Fas-Fas ligand (FasL) interactions play a significant role in the immune privilege status of certain cell populations, and several cytokines and growth factors can modulate their expression. When a FasL-expressing cell binds a Fas bearing immune cell, it triggers its death by apoptosis. In this study, we demonstrate that normal human endometrial epithelial but not stromal cells express FasL. Moreover, we showed that macrophage-conditioned media induced FasL expression by endometrial stromal cells in a dose-dependent manner. To elucidate which macrophage product was responsible for the up-regulation of FasL, endometrial stromal cell cultures were treated with the macrophage products platelet-derived growth factor (PDGF), transforming growth factor (TGF)-beta1, and basic fibroblast growth factor (bFGF). The first two (which are known to be elevated in the peritoneal fluid of women with endometriosis) induced a dose dependent up-regulation of FasL expression, which was specifically inhibited by the antibody. Interestingly, bFGF (which is not elevated in peritoneal fluid of women with endometriosis) did not induce any response. These results suggest that the pro-inflammatory nature of the peritoneal fluid of women with endometriosis induces the FasL expression by regurgitated endometrial cells, and signals Fas mediated cell death of activated immune cells. This could be a mechanism for endometrial cells to escape immune surveillance, implant and grow. PMID- 10381820 TI - Oestrogen receptor (ER)-alpha and ER-beta isoforms in normal endometrial and endometriosis-derived stromal cells. AB - Several investigators have noted that hormone-dependent development of endometriosis implants lags behind that of simultaneously analysed eutopic endometrium. With the recent discovery of the oestrogen receptor-beta (ER-beta) isoform, the aim of this study was to investigate whether differences in the expression of ER-alpha and ER-beta might explain this observation. mRNA transcripts from endometrial stromal cells isolated from normal endometrium (NE) and from endometriomas (EI) were analysed using a semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) technique. RT-PCR and Southern blot analyses of the two ER isoforms indicated that NE and EI stromal cells predominantly express ER-alpha mRNA, however the relative concentrations of ER isoform mRNA transcripts differed between the two cell types. Steady-state ER alpha:ER-beta mRNA ratios were 15.5 +/- 2.8 and 5.2 +/- 0.9 respectively for NE and EI cells (P = 0.02). NE and EI stromal cells expressed ER proteins with similar Kd ( approximately 0.9 nM) and densities ( approximately 24 500 binding sites/cell) respectively. Functional ER expression was indicated by an increase in progesterone receptor concentrations of approximately 60% (P = 0.03) after incubation with 10 nM oestradiol. We postulate that differential transcript processing, ligand specificity and biological actions of the ER-alpha and -beta isoforms may influence differential growth responses in normal and ectopic endometrium. PMID- 10381818 TI - Possible involvement of arylamine N-acetyltransferase 2, glutathione S transferases M1 and T1 genes in the development of endometriosis. AB - Wide inter-individual variation of expression of compound metabolic enzymes is determined by polymorphism and may predispose the development of diseases provoked by environmental factors. The combined analysis of phase II detoxification system genes: arylamine N-acetyltransferase 2 (NAT2), and glutathione S-transferases (GST) M1 and T1 was carried out in patients with minimal/mild (group I; n = 36) and moderate/severe endometriosis (group II; n = 29) and controls (n = 72) of French origin, using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). The results show a significant difference between patients and controls with regard to NAT2 gene polymorphism (P < 0.05). This is mainly due to the high percentage of slow acetylator genotypes (SA) in patients compared with controls (60.0 versus 38.9%; P < 0.02) with a distinct preponderance in subjects with minimal/mild endometriosis (69.4%, P < 0.005) where there is a significantly elevated frequency of slow allele S1 (NAT2*5) (P = 0.05). Significantly increased proportions of GSTM1-deficient genotypes were found in both groups of patients, in comparison with the controls (75.0 and 79.3% versus 45.8%; P < 0. 0001). A preponderance of GSTT1-negative subjects among patients was also detected, but did not appear significant. We suggest the involvement of both NAT2 and GSTM1 detoxification system genes in the pathogenesis of endometriosis and the possible impact of NAT2 gene polymorphism in the development of different forms of this disease. PMID- 10381821 TI - Secretory leukocyte protease inhibitor (SLPI) concentrations in cervical mucus of women with normal menstrual cycle. AB - Secretory leukocyte protease inhibitor (SLPI) is a potent inhibitor of human leukocyte elastase. SLPI transcripts in the cervical tissue were detected during the menstrual cycle by reverse transcription-polymerase chain reaction (RT-PCR). Western blot analysis revealed that the intensity of SLPI protein in cervical tissue in the ovulatory phase was stronger than in other phases. Immunohistochemistry using an anti-SLPI polyclonal antibody revealed positive staining in the epithelial cells of the endocervix. Western blot analysis also revealed that SLPI protein was present in the cervical mucus. Again the intensity of SLPI protein in the ovulatory phase was stronger than that in the follicular phase. The SLPI concentrations and SLPI/elastase ratios in the cervical mucus of women in the ovulatory phase were significantly higher than in the follicular and luteal phases. The SLPI and elastase concentrations in the cervical mucus were positively correlated. No significant difference was found in the SLPI serum concentrations of women during the menstrual cycle. These results suggest that production of SLPI from cervical epithelial cells during the ovulatory phase may be important for protection from the effects of elastase. PMID- 10381822 TI - Urinary trypsin inhibitor down-regulates hyaluronic acid fragment-induced prostanoid release in cultured human amnion cells by inhibiting cyclo-oxygenase-2 expression. AB - We postulated that urinary trypsin inhibitor (UTI), a Kunitz-type protease inhibitor, may inhibit low molecular weight hyaluronic acid (HA) fragment-induced prostanoid release and de-novo expression of the inducible cyclo-oxygenase-2 (COX 2) isoform in human term amnion cells. Purified amnion cultures were obtained from human fetal membranes and were exposed to a HA fragment (molecular weight 35 kDa) in the presence or absence of UTI (0-5.0 micromol/l). Amnion cells treated with the HA fragment (100 nmol/l) released significantly more prostanoids (PGE2 and PGF2alpha) than controls (PGE2: 2.1 +/- 0.13 pg/10(6) cells/24 h compared with 0.42 +/- 0.01, P < 0.05; PGF2alpha: 1.0 +/- 0.17 pg/10(6) cells/24 h compared with 0.13 +/- 0.01, P < 0.05). UTI inhibited HA fragment-induced prostanoid release in a dose-dependent manner, with 50% inhibitory concentration values of 0.8 micromol/l for PGE2 and 1.9 micromol/l for PGF2alpha. Western blot analyses demonstrated that protein levels of COX-2 were substantially increased in amnion cells treated with HA fragment. HA fragment-mediated COX-2 production was markedly diminished by pretreatment with UTI (1.0 micromol/l). These results are the first to demonstrate that UTI is a potent inhibitor of HA fragment induced arachidonic acid metabolism. PMID- 10381823 TI - Expression of 5-lipoxygenase and 5-lipoxygenase-activating protein in human fetal membranes throughout pregnancy and at term. AB - Lipoxygenase metabolites may be involved in human parturition. 5-lipoxygenase (5 LOX) catalyses the first steps in the synthesis of leukotrienes from arachidonic acid, and its activity is dependent on 5-LOX activating protein (FLAP). The expression of 5-LOX and FLAP were investigated in fetal membranes to determine whether there are changes with gestational age or at term with the onset of labour. No significant differences were found in the expression of 5-LOX or FLAP mRNA in the amnion at different gestational ages or at term. In the chorion decidua, 5-LOX mRNA expression was significantly higher in the first trimester of pregnancy than in the second and third trimesters. At term, there was a significant increase in both 5-LOX mRNA and protein expression in the chorion decidua in the time after labour, compared with the time before labour. The expression of FLAP mRNA was also significantly higher in the chorion-decidua in the first trimester of pregnancy compared with the third trimester, and at term in the time after labour compared with the time before labour. Expression of FLAP protein was not studied, as an antibody is not currently available. These results are consistent with a role for 5-LOX and FLAP in the control of parturition at term, and also suggest an involvement earlier in pregnancy. PMID- 10381824 TI - Production and characterization of monoclonal antibodies against pregnancy associated plasma protein A. AB - Pregnancy-associated plasma protein A (PAPP-A) was found to be a good first trimester maternal serum marker, together with free beta-human chorionic gonadotrophin (HCG) subunits, for the biochemical screening of fetal trisomy 21 (Down's syndrome). We have raised monoclonal antibodies (mAbs) against PAPP-A purified from human pregnancy serum. The different antibodies were characterized biochemically by Western blot analysis and in terms of specificity (reaction with non-pregnant and male serum). Their performance in Down's syndrome screening was assessed in comparison with an existing enzyme-linked immunosorbent assay method after labelling of the different mAbs with biotin or horseradish peroxidase. A pair of mAbs was eventually chosen for a double-antibody sandwich protocol. The selected combination was found to have a significantly increased specificity (P = 0.0116) over the method using (purified) polyclonal antibodies, together with slightly increased sensitivity. In our limited number of Down's syndrome pregnancy samples (n = 17) and controls (n = 18), the medians as well as the multiples of the median values (for the affected cases) were comparable between the two methods described. PMID- 10381825 TI - Preimplantation genetic diagnosis and sperm analysis by fluorescence in-situ hybridization for the most common reciprocal translocation t(11;22). AB - In this study we describe the pre-clinical development and clinical application of preimplantation genetic diagnosis (PGD) by fluorescence in-situ hybridization (FISH) for two non-related carriers (one male and one female) of the most common balanced reciprocal translocation: t(11;22)(q25;q12). For the couple with the female carrier, enumeration of the sex chromosomes in the embryos was also indicated (husband: 47,XXY karyotype). Four-colour FISH analysis was performed on six blastomeres from three embryos. No embryo transfer was possible because all the embryos were unbalanced. Three PGD cycles, with two-colour FISH, were carried out for the couple with the male translocation carrier. A total of 35 embryos were biopsied and diagnosed by FISH; nine out of the 35 embryos (25. 7%) were normal and seven of them were transferred (two embryos from the first and four from the third cycle), six out of 35 embryos (17%) were unbalanced, three out of 35 embryos (5.7%) were triploid or polyploid, 10 out of 35 embryos (28.6%) were mosaic and seven out of 35 embryos (20%) were chaotic. Diagnosis failed in 2.9% of the embryos. The spermatozoa of the male carrier were also analysed using three-colour FISH. Only 29.1% of the sperm cells seemed to be balanced or normal. By choosing probes lying on both sides of the breakpoints and by using a combination of sub-telomeric or locus-specific probes and centromeric probes, the use of three-colour FISH enabled detection of all the imbalances in sperm and/or cleavage-stage embryos in the patients. This may improve risk assessment and genetic counselling in the future for translocation carriers. PMID- 10381826 TI - Fluorescent PCR and automated fragment analysis in preimplantation genetic diagnosis for 21-hydroxylase deficiency in congenital adrenal hyperplasia. AB - Congenital adrenal hyperplasia (CAH) is an autosomal recessive disease which is most often caused by a deficiency in steroid 21-hydroxylase. The disease is characterized by a range of impaired adrenal cortisol and aldosterone synthesis combined with an increased androgen synthesis. These metabolic abnormalities lead to an inability to conserve sodium and virilization of females. The most common mutation causing the severe form of CAH is a conversion of an A or C at nucleotide (nt) 656 to a G in the second intron of the steroid 21-hydroxylase gene (CYP21) causing aberrant splicing of mRNA. A couple was referred to our centre for preimplantation genetic diagnosis (PGD) for 21-hydroxylase deficiency in CAH. A PGD was set up to detect the nt656 A/C-->G mutation using fluorescent polymerase chain reaction (PCR) and subsequent restriction enzyme digestion and fragment analysis on an automated sequencer. Using DNA or single cells from the father, the normal allele could not be amplified. Non-amplification of the normal allele has been previously described in asymptomatic carriers, therefore the PCR was further developed using heterozygous lymphoblasts from the mother. The PCR was shown to be highly efficient (96% amplification), accurate (0% contamination) and reliable (0% allelic drop-out). The couple started PGD treatment and the second PGD cycle resulted in a twin pregnancy. The genotype of the fetuses was determined in our laboratory using chorionic villus sampling material using the method described here. Both fetuses were shown to be heterozygous carriers of the mutation, and two healthy girls were born. PMID- 10381827 TI - Genocide by Attrition 1939-1993: The Warsaw Ghetto, Cambodia, and Sudan. AB - Genocide by attrition occurs when a group is stripped of its human rights, political, civil and economic. This leads to deprivation of conditions essential for maintaining health, thereby producing mass death. Genocide by attrition is epitomized by the Warsaw Ghetto (1939-43), Democratic Kampuchea (1975-79), and Sudan (1983-93). Potentialities of response are considered, as well as state and international interests in overlooking genocide, and the inadequacy and misuses of humanitarian aid. Lastly, guidelines are offered for future policy to prevent genocide by attrition, involving governments, health professionals, and aid workers. PMID- 10381828 TI - Health and the Environment: A Human Rights Agenda for the Future. AB - Linkages between health and the environment are increasingly recognized, but human rights law still does not provide an adequate framework for dealing with those connections. Environmental health issues now are seen as involving many institutions, processes, actors, and causes that are not immediately obvious. This reality requires changes in the human rights agenda. Practitioners should not take an overly legalistic and norm-based approach to health and the environment. While the right to a healthy environment is a useful concept, it is more effective to focus on how to implement the right to health from a policy perspective. Practitioners need to think about how human rights approaches can pressure decision-makers and social institutions to consider the environmental causes of ill health. Finally, it is hoped that human rights and environmental movements will work together more effectively in the future. PMID- 10381829 TI - The Links between Legal Status and Environmental Health: A Case Study of Mozambican Refugees and Their Hosts in the Mpumalanga (Eastern Transvaal) Lowveld, South Africa. AB - This article relates the legal status of Mozambicans in South Africa from 1985 onwards to key findings of a demographic census taken in 1992, an environmental health survey conducted in 1993, and in-depth fieldwork in some of the surveyed settlements in 1995. The case study area on the border with Mozambique is typical of South Africa's rural former homelands, with the exception that it has a large and long-standing refugee population. The environmental health indicators for refugees are considerably worse than for their hosts, and in-depth fieldwork suggests that this can be attributed to their legal and political vulnerability. This raises issues for South Africa's Reconstruction and Development Program, as well as conceptual challenges for promoters of human rights. PMID- 10381830 TI - The Next Plague: Stigmatization and Discrimination Related to Hepatitis C Virus Infection in Australia. AB - A recently discovered hepatitis C virus is a common cause of chronic liver disease in industrialized countries. Because it is basically blood-borne and because blood donors are systematically screened, the only major group now at risk of infection are injecting drug users. There are increasing reports of stigmatization, affronts to dignity and discrimination as a result of the hepatitis C status of individuals, but little action is being taken to prevent or redress these. In an attempt to stimulate such action, we collected 37 reports of such incidents in Australia in 1994, in the domestic, work, recreational, day care and funeral settings, but the most common involved health care settings and health professionals. In general, action did not follow from such incidents, despite the fact that Australia has a very strong framework of anti discrimination legislation and process. It is urgently necessary that these issues be addressed, both in themselves and as a necessary prerequisite for controlling the continued massive spread of hepatitis C among injecting drug users. PMID- 10381831 TI - Juveniles and Psychiatric Institutionalization: Toward Better Due Process and Treatment Review in the United States. AB - The United Nations Convention on the Rights of the Child (CRC) can be used as a framework to examine issues regarding psychiatric institutionalization of juveniles in the United States. The current U.S. system allows children diagnosed with relatively mild, non-psychotic disorders or exhibiting delinquent behaviors to be placed in institutions. Failure to regulate treatment in these faciltities also results in abuses by the treatment providers. Parents can institutionalize a child under the guise of mental health "treatment" because they disapprove of the child's lifestyle choices. In some states, parents can waive the child's right to an impartial hearing before institutionalization. The serious social, mental, and physical health consequences of erroneous deprivation of liberty are discussed. Recommendations include that the U.S. ratify the CRC, guarantee due process for juveniles faced with institutionalization, conduct systematic treatment reviews, and correct institutional abuses. PMID- 10381832 TI - Haiti 1991-1994: The International Civilian Mission's Medical Unit. PMID- 10381833 TI - The Relevance of Human Rights to Health Status in Australiam Aboriginal and Torres Strait Islander Communities. PMID- 10381834 TI - The synergistic relationship between health and human rights: a case study using female genital mutilation. PMID- 10381835 TI - Chromosome aberrations of clonal origin in irradiated and unexposed individuals: assessment and implications. AB - Chromosome painting has proven useful for the detection of chromosomal rearrangements, although the presence of cells containing clonal aberrations can have an effect on the outcome of cytogenetic analyses (e.g. aberration frequency and chromosomal distribution studies). Cells with clonal chromosomal changes have been found in studies of both radiation-exposed Chernobyl cleanup workers ("liquidators") and healthy unexposed human subjects. We have used a simple statistical method to aid in the identification of individuals from distinct Chernobyl radiation-exposed and unexposed control populations who may possess cells containing clonal rearrangements. A chi2 value determined from the observed number of aberrations and the expected number based on chromosome length that corresponds to a probability less than 0.005 appears to be an indicator of clonality. These selected individuals can be analyzed further for clonality, thereby sparing detailed examination of the entire population. Here we present an analysis of individuals possessing clonal aberrations to assess the influence of clonality on the results of cytogenetic studies. Our results show that the subtraction of clonal events from the chi2 calculation for the "outliers" results in nearly all of these values losing their statistically significant deviation from proportionality. These adjustments can also be made to prevent the overestimation of frequencies of chromosome aberrations for biodosimetry. The frequency of clonal aberrations appears to increase as a function of age in control subjects, whereas an age effect was not evident in Chernobyl liquidators. This suggests that spontaneous and radiation-induced clonal expansion are occurring in control subjects and liquidators, respectively. PMID- 10381836 TI - Accumulation of P-selectin in the lumen of irradiated blood vessels. AB - Ionizing radiation induces the inflammatory response in part through leukocyte binding to cell adhesion molecules that are expressed on the vascular endothelium. We studied the effects of X radiation on the pattern of immunohistochemical staining of CD62P (P-selectin). P-selectin was localized within cytoplasmic granules in the untreated vascular endothelium. Immunohistochemical staining of P-selectin was observed at the luminal surface of vascular endothelium within 1 h of irradiation. Increased P-selectin staining at the blood-tissue interface occurred primarily in pulmonary and intestinal blood vessels. To determine whether localization of P-selectin at the vascular lumen occurs through exocytosis of endothelial cell stores in addition to platelet aggregation, we removed the vascular endothelium from the circulation and irradiated endothelial cells in vitro. In this system, we studied the mechanisms by which ionizing radiation induced translocation of P-selectin by using immunofluorescence of human umbilical vein endothelial cells (HUVEC) and confocal microscopy. Prior to irradiation, P-selectin is localized in cytoplasmic reservoirs of HUVEC. After irradiation of HUVEC, P-selectin was translocated to the cell membrane, where it remained tethered. The lowest dose at which we could expect translocation of P-selectin to the cell membrane was 2 Gy. To determine whether P-selectin in Weibel-Palade bodies requires microtubule-dependent membrane transport, we added two microtubule-depolymerizing agents, Colcemid and nocodazole. Microtubule-depolymerizing agents prevented radiation-induced trans- location of P-selectin to the cell membrane. Thus P-selectin accumulates in irradiated blood vessels through both platelet aggregation and microtubule dependent exocytosis of storage reservoirs within the vascular endothelium. PMID- 10381837 TI - The role of laminin-1 in the modulation of radiation damage in endothelial cells and differentiation. AB - Microvascular dysfunction due to endothelial damage is often associated with the ionizing radiation used during cancer therapy. This radiation-induced capillary injury is a major factor in the inhibition of new vessel growth (angiogenesis) and in disease states such as radiation-induced pneumonitis and nephropathy. Many studies have examined the effects of radiation on endothelial cell function; however, little is known regarding the role the basement membrane plays in radiation-induced endothelial cell damage and angiogenesis. Therefore, we examined the effects of gamma radiation on aortic explants, and in vitro on three endothelial cell types (of artery, vein and capillary origin) irradiated with or without the basement membrane glycoprotein laminin-1. As expected, irradiation inhibited angiogenic sprouting of the aortic explants, endothelial cell proliferation, attachment, migration and differentiation in vitro in a dose dependent manner. However, the effect of radiation on several of these processes in angiogenesis was reduced when the cells were irradiated on laminin-1. To further evaluate the effects of radiation on endothelial cells, we examined the expression of the vascular endothelial cell growth factor (VEGF) kinase domain region receptor in endothelial cells irradiated in the presence and absence of laminin-1. In endothelial cells irradiated on laminin-1, KDR expression increased 2.5-fold over control levels. Therefore, although radiation has a dose-dependent inhibitory effect on processes associated with angiogenesis in vitro, the presence of the basement membrane glycoprotein laminin-1 during irradiation decreases these effects. PMID- 10381838 TI - Biology of marrow stromal cell lines derived from long-term bone marrow cultures of Trp53-deficient mice. AB - To investigate the effect of Trp53 (formerly known as p53) on stromal cells of the hematopoietic microenvironment, long-term bone marrow cultures were established from mice in which the Trp53 gene had been inactivated by homologous recombination (Trp53(-/-)) or their wild-type littermates (Trp53(+/+)). Long-term bone marrow cultures from Trp53(-/-) mice continued to produce nonadherent cells for 22 weeks, while Trp53(+/+) cultures ceased production after 15 weeks. There was a significant increase in the number of nonadherent cells produced in Trp53( /-) long-term bone marrow cultures beginning at week 9 and continuing to week 22 (P < 0.02). The Trp53(-/-) cultures also showed significantly increased cobblestone island formation indicative of early hematopoietic stem cell containing colonies beginning at week 10 (P < 0.01). Cobblestone islands persisted until weeks 15 and 22 in Trp53(+/+) and Trp53(-/-) cultures, respectively. Co-cultivation experiments in which Trp53(+/+) Sca1(+)lin- enriched hematopoietic stem cells were plated on Trp53(-/-) stromal cells showed increased cobblestone island formation compared to Trp53(-/-) Scal+lin- cells plated on Trp53(+/+) or Trp53(-/-) stromal cells. Radiation survival curves for clonal bone marrow stromal cells revealed a similar D0 for the Trp53(+/+) and Trp53(-/-) cell lines (1.62 +/- 0.16 and 1.49 +/- 0. 08 Gy, respectively; P = 0.408), and similar n (8.60 +/- 3.23 and 10.71 +/- 0.78, respectively) (P = 0.491). Cell cycle analysis demonstrated a G2/M-phase arrest that occurred 6 h after irradiation for both Trp53(+/+) and Trp53(-/-) stromal cell lines. After 10 Gy irradiation, there was no significant increase in the frequency of apoptosis detected in Trp53(+/+) compared to Trp53(-/-) marrow stromal cell lines. In the stromal cell lines, ICAM 1 was constitutively expressed on Trp53(+/+) but not Trp53(-/-) cells; however, a 24-h exposure to TNF-alpha induced detectable ICAM-1 on Trp53(-/-) cells and increased expression on Trp53(+/+) cells. To test the effect of Trp53 on the radiation biology of hematopoietic progenitor cells, the 32D cl 3 cell line was compared with a subclone in which expression of an E6 inserted transgene accelerates ubiquitin-dependent degradation of Trp53, thus preventing accumulation of Trp53 after genotoxic stress. The radiation survival curves were similar with no significant difference in the D0 or n, or in the percentage of cells undergoing apoptosis after 10 Gy irradiation between the two cell lines. Cells of the 32D-E6 cell line displayed a G2/M-phase arrest 6 h after 10 Gy, while cells of the parent line exhibited both a G2/M-phase arrest and a G1-phase arrest at 24 and 48 h. The results suggest a complex mechanism of action of Trp53 on the interactions between stromal and hematopoietic cells in long-term bone marrow cultures. PMID- 10381839 TI - In vivo electrical impedance spectroscopic monitoring of the progression of radiation-induced tissue injury. AB - This study evaluates the potential of electrical impedance spectroscopy (EIS) as a noninvasive technique for tracking the progression of radiation-induced damage in normal muscle tissue. Male Sprague-Dawley rats were irradiated locally to the gastrocnemius and biceps femoris muscle. Single doses were administered using a procedure that spares skin and bone. Complex impedance spectral measurements (taken at 50 frequency points between 1 kHz and 1 MHz) were made at monthly intervals using recessed disk electrodes applied to the skin. A histological scoring scheme was developed for evaluation of injury. A strong dose-dependent progression of injury evident in both spectral measurements and histological scoring has been observed. Latent time also appears to be dependent on dose with changes induced by 70 Gy evident by 2 months, changes induced by 90 Gy observed by 1 month, and dramatic changes found within 3 weeks at 150 Gy. Injury was morphologically comparable to the type of damage that occurs in response to small, fractionated doses, but on a much shorter time scale. Increased spectral shift was a consistent indicator of the extent of tissue injury at the time of measurement. The use of a large single dose resulted in an excellent model in terms of inducing a significant progression in tissue injury over a short post treatment follow-up period in the muscle mass while also providing a consistent location for in vivo electrical impedance measurements. The results show that EIS can follow radiation-induced tissue change, suggesting that EIS has the potential to monitor the types of injury observed in late radiation damage of muscle tissue noninvasively. PMID- 10381840 TI - Hormone pretreatment enhances recovery of spermatogenesis in rats after neutron irradiation. AB - Previous studies showed that a 6-week pretreatment of rats with testosterone plus estradiol enhanced the recovery of spermatogenesis 9 weeks after gamma irradiation, resulting in a dose-modifying factor (DMF) of about 2. To test whether the effect of the hormone treatment was mediated through changes in oxygen tension, thiol levels or DNA repair, we irradiated the testes of rats with neutrons, which depend less on these factors than does low-LET radiation for their cytotoxic action. Control rats and rats treated with testosterone plus estradiol were irradiated with 0.7-2.7 Gy of cyclotron-generated high-energy neutrons. The recovery of spermatogenesis was assessed 9 weeks after irradiation by testis weights, sperm counts and the tubule repopulation indices. Greater recovery of spermatogenesis was observed for all end points, with a DMF of about 2 for rats treated with testosterone plus estradiol compared to the irradiated, cholesterol-treated rats. The equal protection factors for neutrons and gamma rays indicate that oxygen, thiols and repair of DNA damage are unlikely to be involved in the protective effect of the hormone treatment. PMID- 10381842 TI - Effect of gamma radiation on native endolithic microorganisms from a radioactive waste deposit site. AB - A time-course experiment was conducted to evaluate the effects of gamma radiation on the indigenous microbiota present in rock obtained from Yucca Mountain, Nevada Test Site. Microcosms were constructed by placing pulverized Yucca Mountain rock in polystyrene cylinders. Continuous exposure (96 h) at a dose rate of 1.63 Gy/min was used to mimic the near-field environment surrounding waste canisters. The expected maximum surface dose rate from one unbreached canister designed to contain spent nuclear fuels is 0.06 Gy/min. Considering the current repository packing design, multiple canisters within one vault, the cumulative dose rate may well approach that used in this experiment. The microbial communities were characterized after receiving cumulative doses of 0, 0.098, 0. 58, 2.33, 4.67, 7.01 and 9.34 kGy. Radiation-resistant microorganisms in the pulverized rock became viable but nonculturable (VBNC) after a cumulative dose of 2.33 kGy. VBNC microorganisms lose the ability to grow on media on which they have routinely been cultured in response to the environmental stress imposed (i.e. radiation) but can be detected throughout the time course using direct fluorescence microscopy techniques. Two representative exopolysaccharide-producing isolates from Yucca Mountain were exposed to the same radiation regimen in sand microcosms. One isolate was much more radiation-resistant than the other, but both had greater resistance than the general microbial community based on culturable counts. However, when respiring cell counts (VBNC) were compared after irradiation, the results would indicate much more radiation resistance of the individual isolates and the microbial community in general. These results have significant implications for underground storage of nuclear waste as they indicate that indigenous microorganisms are capable of surviving gamma irradiation in a VBNC state. PMID- 10381841 TI - Alpha particles induce the production of interleukin-8 by human cells. AB - The pulmonary microenvironment is a primary target for alpha particles like those emitted by inhaled radon and its progeny. While exposure to alpha particles has recently been associated with the generation of extracellular and intracellular reactive oxygen species (ROS; Cancer Res. 57, 3963-3971, 1997), little is known about how exposure to alpha particles may affect the generation of oxidative stress-related mediators in the respiratory tract. Interleukin-8 (IL8) is a cytokine recognized for its potent role as a chemoattractant and activator of polymorphonuclear leukocytes. Oxidative stress can up-regulate expression of the gene that encodes IL8 (IL8) in a variety of cell types. In this study, we set out to investigate a potential linkage between the generation of ROS and production of IL8 in alpha-particle-irradiated normal human lung fibroblasts. ELISA revealed that exposure of the fibroblasts to low doses of alpha particles (3.6-19 cGy) caused significant increases in generation of the IL8 protein as early as 30 min after irradiation. Northern blot analyses revealed that such increases were associated with increased IL8 mRNA levels. Cells exposed to alpha particles in the presence of antioxidants, i.e. superoxide dismutase and dimethyl sulfoxide, resulted in significant decreases in extracellular IL8 protein levels. Similar results were obtained with cells treated with dexamethasone, an inhibitor of transcription. Our results indicate that alpha-particle-induced increases in production of IL8 occur temporally in parallel with elevated production of ROS. Conceivably, such production of IL8 induced by alpha particles may contribute to an inflammatory response in the lower respiratory tract. Additionally, the promitogenic effects of IL8 may be a factor in hyperplastic responses in the airway epithelial cells to inhaled radon and radon progeny and perhaps other stresses associated with ROS. PMID- 10381843 TI - Resuscitation of microorganisms after gamma irradiation. AB - Microbiological analysis of rock exposed to gamma-radiation doses between 0 and 9.34 kGy indicated that some microorganisms became viable but nonculturable (VBNC) and lost metabolic capacity as measured by BIOLOG microtiter plates. To investigate this phenomenon, portions of irradiated rock were placed at 4 degrees C for 2 months in an attempt to resuscitate the microbes to a culturable state. Culturable heterotrophs were enumerated and BIOLOG plates were used to determine the metabolic capability of the microbial community. Culturable bacteria that had previously been nonculturable were found at all doses. The number of colony types decreased from 26 in the nonirradiated control rock to between 9 and 10 in rock irradiated at doses ranging from 2.34 to 9.34 kGy. BIOLOG plates indicated partial recovery of metabolic capacity in all the samples tested. Fatty acid methyl ester analysis of the recovered isolates using the MIDI system (Microbial ID, Inc.) yielded three distinct groups of related bacteria. All resuscitated isolates clustered with the original nonirradiated isolates at the genus level, and 92% of them clustered at the species level. These results indicate that microbes were likely resuscitated from a VBNC state. PMID- 10381845 TI - The radiolysis and radioracemization of poly-L-leucines. AB - The brief history of the discovery of radioracemization, the racemization of an optically active substance induced by ionizing radiation, is reviewed. Our early studies involving the radiolysis and radioracemization of D- and L-leucine using gamma radiation from a 111-TBq 60Co gamma-ray source are described briefly, as are later experiments involving other protein amino acids and their salts, as well as the nonprotein amino acid, isovaline. The implications of the results of such studies for the Vester-Ulbricht mechanism which proposes longitudinally polarized beta radiation as the origin of molecular chirality, for the cosmological question of the enantiomeric compositions of amino acids in the Murchison meteorite, and for the use of D/L ratios of amino acids for geochronological and geothermal estimates are reviewed briefly. These past radiolysis-radio- racemization studies have involved only monomeric amino acids. The present research, extending such investigations to two homochiral L-leucine polypeptides, (L-Leu)10 and (L-Leu)78, was undertaken to see if a polymer of an amino acid might be more stable to radiolysis and radioracemization than the corresponding monomer. It was found that these polypeptides were more stable to radiolysis than was the leucine monomer, but that the extents of radioracemization in all samples were comparable. PMID- 10381844 TI - Measurement of dose rate at the interface of cell culture medium and glass dishes by means of ESR dosimetry using thin films of alanine. AB - Previous studies on human cervical cancer cells (NHIK 3025) have indicated that the cells, when X-irradiated in suspension, appeared to be more radiosensitive than when they were irradiated attached to glass dishes. However, this result depends on dosimetry, which is difficult in the situation where cells are attached to glass dishes due to backscattering electrons at the glass-liquid interface. Recently developed dosimetry that is based on detection of radiation induced stable radicals in alanine and uses ESR spectroscopy offers a possibility for more relevant dosimetry at the glass-liquid interface than the previous estimates of doses based on Fricke dosimetry. Thin alanine films (>/=10 microm) were used to measure dose at the interface by irradiating the films while they were placed tightly against the bottom of dishes and covered with 1 mm of wax simulating the medium above cells. Fricke dosimetry was also performed, with different depths of Fricke solution in the dishes, to elucidate the contribution to the dose delivered by backscattering electrons at the glass-liquid interface. A dose rate of 1.9 Gy/min was measured with a thin layer (0.2-0.3 mm) of Fricke solution in petri dishes made of glass. However, this estimate appears to be too high, due to a contribution to dose by short-ranged electrons generated when the X rays passed through a steel lid 4.5 cm above the dishes. Dosimetry using alanine films resulted in dose rates of 1.15 and 0.87 Gy/min at the interfaces of glass-liquid and plastic- liquid, respectively. Hence there is a significant contribution to dose from backscattering electrons on dishes made of glass. The reason for our previous observation of a difference in radiosensitivity between cells irradiated in suspension and cells irradiated attached to glass appears to be a lack of accurate dosimetry at the glass-liquid interface. PMID- 10381846 TI - Randomized trials in cardiogenic shock: what's the problem? PMID- 10381847 TI - The power of opinion polls. PMID- 10381849 TI - Is a low fat diet enough to achieve serum cholesterol goals? PMID- 10381848 TI - Soluble adhesion molecules in ischaemic heart disease. PMID- 10381850 TI - The acid test: Na+/H+ exchange and its inhibitors in acute myocardial ischaemia. PMID- 10381851 TI - AT1-receptor blockers in hypertension and heart failure: clinical experience and future directions. PMID- 10381852 TI - Does stent design influence restenosis? PMID- 10381853 TI - Indications for intracoronary stent placement: the european view. Working Group on Coronary Circulation of the European Society of Cardiology. AB - AIMS: In Europe, no written official guidelines on indications for coronary stent placement are available. We therefore assessed the opinions of European interventional cardiologists on these indications. METHODS AND RESULTS: In April 1997, a questionnaire was sent to the members of the Working Group on Coronary Circulation of the European Society of Cardiology with interventional cardiology as the main activity. A total of 165 questionnaires were returned and analysed. For the treatment of acute or threatened vessel closure during PTCA, the threshold for stenting is set at the level of a type C dissection by 42% of the cardiologists, while 22% stent any form of dissection and 13% require an impaired TIMI flow. A suboptimal PTCA result necessitating stenting is defined as a residual stenosis of >50% by 35% or of 30 >30% by 55% of the respondents. When considering primary prevention of restenosis, only 2% unconditionally stent focal, new-onset lesions in native coronary arteries, 44% refrain from stenting in a case of a stent-like PTCA result (3.37mmol x l-1 were investigated 12-14 weeks after an acute coronary event. After overnight fasting each patient had (a) his resting energy expenditure measured (indirect calorimetry using standard protocol) and (b) venous blood sampled from a forearm vein to determine lipid profile. All the patients were randomly allocated to four groups of treatment: Group A on a very low fat diet (resting energy expenditure-fat diet, where fat intake was 60% of the cells and the value increased significantly after MIP. In the presence of 5x10(-6) mol/L nor-binaltorphimine (nor-BNI), a selective kappa-OR antagonist, but not 5x10(-6) mol/L CTOP, a selective mu-OR antagonist, or 5x10(-6) mol/L naltrindole, a selective delta-OR antagonist, the cardioprotection of MIP was significantly attenuated. To verify the role of kappa-OR, we studied the effects of severe MI after pretreatment with the kappa-OR agonist U50,488H (UP) for 30 minutes. U50,488H at 3x10(-6) to 1x10(-4) mol/L increased cell viability concentration dependently with an EC50 of 3.311x10(-6) mol/L. In the presence of 5x10(-6) nor BNI, the cardioprotection of UP (3x10(-5) mol/L) was blocked. A time course study showed that UP-induced cardioprotection occurred in 2 windows: the first occurred approximately 1 hour later and the other occurred 16 to 20 hours later. Additional studies on cell contraction and intracellular Ca2+ ([Ca2+]i) revealed that both UP and MIP attenuated the inhibitory effects of severe MI on contractility and electrically induced [Ca2+]i transient in single ventricular myocytes. On blockade of protein kinase C, the delayed cardioprotections of UP and MIP were significantly attenuated. In conclusion, the results of the present study have provided evidence that kappa-OR mediates the cardioprotection of MIP, which may involve protein kinase C and [Ca2+]i. PMID- 10381892 TI - Intracellular sodium accumulation during ischemia as the substrate for reperfusion injury. AB - To elucidate the role of intracellular Na+ kinetics during ischemia and reperfusion in postischemic contractile dysfunction, intracellular Na+ concentration ([Na+]i) was measured in isolated perfused rat hearts using 23Na nuclear magnetic resonance spectroscopy. The extension of the ischemic period from 9 minutes to 15, 21, and 27 minutes (at 37 degrees C) increased [Na+]i at the end of ischemia from 270.0+/-10.4% of preischemic level (mean+/-SE, n=5) to 348.4+/-12.0% (n=5), 491.0+/-34.0% (n=7), and 505.3+/-12.1% (n=5), respectively, whereas the recovery of developed pressure worsened with the prolongation of the ischemic period (95.1+/-4.2%, 84.3+/-1. 2%, 52.8+/-13.7%, and 16.9+/-6.4% of preischemic level). The kinetics of [Na+]i recovery during reperfusion was analyzed by the fitting of a monoexponential function. When the hearts were reperfused with low-[Ca]o (0.15 mmol/L) solution, the time constants of the recovery (tau) after 15-minute (8.07+/-0.85 minutes, n=5) and 21-minute ischemia (6.44+/-0.90, n=5) were significantly extended, with better functional recovery (98.5+/-1.4% for 15-minute [P<0.05]; 98.0+/-1.0% for 21-minute [P<0.05]) compared with standard reperfusion ([Ca]o=2.0 mmol/L, tau=3.58+/-0.28 minutes for 15 minute [P<0.0001]; tau=3.02+/-0.20 for 21-minute [P<0.0001]). A selective inhibitor of Na+/Ca2+ exchanger also decelerated the [Na+]i recovery, which suggests that the recovery reflects the Na+/Ca2+ exchange activity. In contrast, high-[Ca]o reperfusion (5 mmol/L) accelerated the [Na+]i recovery after 9-minute ischemia (tau=2.48+/-0.11 minute, n=5 [P<0.0001]) and 15-minute ischemia (tau=2.10+/-0.07, n=6 [P<0. 05]), but functional recovery deteriorated only in the hearts with 15-minute ischemia (29.8+/-9.4% [P<0.05]). [Na+]i recovery after 27-minute ischemia was incomplete and decelerated by low-[Ca]o reperfusion, with limited improvement of functional recovery (42. 5+/-7.9%, n=5 [P<0.05]). These results indicate that intracellular Na+ accumulation during ischemia is the substrate for reperfusion injury and that the [Na+]i kinetics during reperfusion, which is coupled with Ca2+ influx, also determines the degree of injury. PMID- 10381893 TI - Contribution of alpha-adrenergic and beta-adrenergic stimulation to ischemia induced glucose transporter (GLUT) 4 and GLUT1 translocation in the isolated perfused rat heart. AB - The intracellular signaling mechanism of the ischemia-stimulated glucose transporter (GLUT) translocation in the heart is not yet characterized. It has been suggested that catecholamines released during ischemia may be involved in this pathway. The purpose of this study was to evaluate the contribution of alpha adrenoceptors and beta-adrenoceptors to ischemia-mediated GLUT4 and GLUT1 translocation in the isolated, Langendorff-perfused rat heart. Additionally, GLUT translocation was studied in response to catecholamine stimulation with phenylephrine (Phy) and isoproterenol (Iso). The results were compared with myocardial uptake of glucose analogue [18F]fluorodeoxyglucose (FDG). Subcellular analysis of GLUT4 and GLUT1 protein on plasma membrane vesicles (PM) and intracellular membrane vesicles (IM) using membrane preparation and immunoblotting revealed that alpha- and beta-receptor agonists stimulated GLUT4 translocation from IM to PM (2.5-fold for Phy and 2.1-fold for Iso, P<0.05 versus control), which was completely inhibited by phentolamine (Phe) and propranolol (Pro), respectively. Plasmalemmal GLUT1 moderately rose after Iso exposure, and this was prevented by Pro. In contrast, ischemia-stimulated GLUT4 translocation (2.2-fold, P<0.05 versus control) was only inhibited by alpha-adrenergic antagonist Phe but not by beta-adrenergic antagonist Pro. Similarly, Phe but not Pro inhibited ischemia-stimulated GLUT1 translocation. GLUT data were confirmed by FDG uptake monitored using bismuth germanate detectors. The catecholamine stimulated FDG uptake (6.9-fold for Phy and 8.9-fold for Iso) was significantly inhibited by Phe and Pro; however, only Phe but not Pro significantly reduced the ischemia-induced 2.5-fold increase in FDG uptake (P<0.05 versus ischemia). This study suggests that alpha-adrenoceptor stimulation may play a role in the ischemia-mediated increase in glucose transporter trafficking leading to the stimulation of FDG uptake in the isolated, perfused rat heart, whereas beta adrenergic activation does not participate in this signaling pathway. PMID- 10381894 TI - Deficient platelet-derived nitric oxide and enhanced hemostasis in mice lacking the NOSIII gene. AB - Endothelial nitric oxide synthase (eNOS) has been identified in human platelets. Although platelet-derived nitric oxide (NO) has been shown to inhibit platelet recruitment in vitro, its role in the regulation of the hemostatic response in vivo has not been characterized. To define the role of platelet-derived NO in vivo, we studied mice that lacked a functional eNOS gene (NOSIII). Surface P selectin expression in platelets from eNOS-deficient mice was not significantly altered; however, bleeding times were markedly decreased in eNOS-deficient versus wild-type mice (77.2+/-3 versus 133.4+/-3 seconds, P<0.00005). To determine the contribution of endothelium- versus platelet-derived NO to the bleeding time, isolated platelets from either eNOS-deficient or wild-type mice were transfused into a thrombocytopenic eNOS-deficient mouse and the bleeding time was measured. The bleeding times in mice transfused with eNOS-deficient platelets were significantly decreased compared with mice transfused with wild-type platelets (Deltableeding time, -24.6+/-9.1 and -3.4+/-5.3 seconds, respectively; P<0.04). Platelet recruitment was studied by measuring serotonin release from a second recruitable population of platelets that were added to stimulated platelets at the peak of NO production. There was 40.3+/-3.7% and 52. 0+/-2.1% serotonin release for platelets added to wild-type or eNOS-deficient platelets, respectively (P<0.05). In summary, mice that lacked eNOS had markedly decreased bleeding times even after endothelial NO production was controlled. These data suggest that the lack of platelet-derived NO alters in vivo hemostatic response by increasing platelet recruitment. Thus, these data support a role for platelet derived NO production in the regulation of hemostasis. PMID- 10381895 TI - Gene transfer of endothelial nitric oxide synthase to the lung of the mouse in vivo. Effect on agonist-induced and flow-mediated vascular responses. AB - The effects of transfer of the endothelial nitric oxide synthase (eNOS) gene to the lung were studied in mice. After intratracheal administration of AdCMVbetagal, expression of the beta-galactosidase reporter gene was detected in pulmonary airway cells, in alveolar cells, and in small pulmonary arteries. Gene expression with AdCMVbetagal peaked 1 day after administration and decayed over a 7- to 14-day period, whereas gene expression after AdRSVbetagal transfection peaked on day 5 and was sustained over a 21- to 28-day period. One day after administration of AdCMVeNOS, eNOS protein levels were increased, and there was a small reduction in mean pulmonary arterial pressure and pulmonary vascular resistance. The pressure-flow relationship in the pulmonary vascular bed was shifted to the right in animals transfected with eNOS, and pulmonary vasodepressor responses to bradykinin and the type V cGMP-selective phosphodiesterase inhibitor zaprinast were enhanced, whereas systemic responses were not altered. Pulmonary vasopressor responses to endothelin-1 (ET-1), angiotensin II, and ventilatory hypoxia were reduced significantly in animals transfected with the eNOS gene, whereas pressor responses to norepinephrine and U46619 were not changed. Systemic pressor responses to ET-1 and angiotensin II were similar in eNOS-transfected mice and in control mice. Intratracheal administration of AdRSVeNOS attenuated the increase in pulmonary arterial pressure in mice exposed to the fibrogenic anticancer agent bleomycin. These data suggest that transfer of the eNOS gene in vivo can selectively reduce pulmonary vascular resistance and pulmonary pressor responses to ET-1, angiotensin II, and hypoxia; enhance pulmonary depressor responses; and attenuate pulmonary hypertension induced by bleomycin. Moreover, these data suggest that in vivo gene transfer may be a useful therapeutic intervention for the treatment of pulmonary hypertensive disorders. PMID- 10381896 TI - Identification and cloning of a secreted protein related to the cysteine-rich domain of frizzled. Evidence for a role in endothelial cell growth control. AB - We report the isolation of a cDNA, FrzA (frizzled in aorta; GenBank accession No. U85945), from bovine aortic endothelium. It is the bovine counterpart of the mouse sFRP1, which encodes for a secreted protein that is homologous to the cysteine-rich domain of frizzled. Members of the frizzled family of genes have been shown to be required for tissue polarity and to act as receptors for Wnt. The predicted protein product of this gene includes the cysteine-rich extracellular domain, but not the 7 putative transmembrane domains that are highly conserved among members of the frizzled family. Visualization of FrzA mRNA and protein revealed that it was widely distributed among adult tissues. FrzA is expressed by highly differentiated or polarized cells, eg, neurons, cardiocytes, or various epithelia. Analysis of its expression in endothelium revealed that FrzA mRNA levels were high in endothelial cells scraped from freshly obtained bovine aortas, decreased when cells were placed in culture and began to proliferate, but increased at confluence. Transient transfection assays and an assay using addition of purified protein indicate that FrzA reduces the proliferation of endothelial cells. These data demonstrate the existence of a secreted protein homologous to the extracellular domain of the fz receptor, which we speculate plays a role in controlling cell growth and differentiation, possibly by regulating accessibility to Wnt family members. PMID- 10381897 TI - Adenovirus-mediated local expression of human tissue factor pathway inhibitor eliminates shear stress-induced recurrent thrombosis in the injured carotid artery of the rabbit. AB - The main cause of acute coronary syndrome may be recurrent thrombosis, which is initiated by the activation of the extrinsic coagulation pathway. Tissue factor (TF) pathway inhibitor (TFPI) efficiently inhibits an early step in this pathway by the formation of a complex with factor VIIa, TF, and factor Xa. We determined whether local TFPI gene transfer can inhibit thrombosis in an injured artery without inducing systemic side effects. Balloon-injured rabbit carotid arteries were infected with an adenoviral vector that expressed either human TFPI (AdCATFPI) or bacterial beta-galactosidase (AdCALacZ). Two to 6 days after gene transfer, thrombosis was induced by the production of constant stenosis of the artery, and blood flow was measured continuously with an electromagnetic flow probe. A cyclic flow variation, which is thought to reflect the recurrent formation and dislodgment of mural thrombi, was observed in all AdCALacZ-infected arteries as well as in saline-infused arteries. In contrast, no cyclic flow variation was detectable in AdCATFPI-transfected arteries, even in the presence of epinephrine (1 microg. kg-1. min-1 infusion). Prothrombin time, activated partial thromboplastin time, and the ex vivo platelet aggregation induced by either adenosine diphosphate or collagen were unaltered in AdCATFPI-infected rabbits. We found that in vivo TFPI gene transfer into an injured artery completely inhibits the recurrent thrombosis induced by shear stress even in the presence of catecholamine, without affecting systemic coagulation status. Adenovirus-mediated local expression of TFPI may have the potential for the treatment of human thrombosis. PMID- 10381898 TI - Cosegregation analysis in genetic crosses suggests a protective role for atrial natriuretic factor against ventricular hypertrophy. AB - In most rat models studied to date, increased ventricular mass is associated with high ventricular expression of the atrial natriuretic factor (ANF) gene. However, it is unknown whether ANF plays a beneficial or detrimental role in the course of left ventricular hypertrophy or whether ANF gene expression could be genetically linked to cardiac mass. To address such questions, we performed a cosegregation analysis in genetic crosses of inbred strains of rats. To select strains with the appropriate phenotypic characteristics, we first compared the ventricular abundance of ANF mRNA to ventricular mass (corrected for body weight) in 2 recombinant inbred strains derived from Wistar-Kyoto (WKY)/spontaneously hypertensive rat (SHR) hybrid crosses, ie, WKY-derived hyperactive (WKHA) and WKY derived hypertensive (WKHT) rats, as well as in their parental inbred strains. In the 2 such strains that were normotensive, we observed that ventricular mass was higher in WKHA than in WKY rats, yet ventricular ANF mRNA was less abundant in WKHA than in WKY rats. Within a segregating population of F2 animals generated from a cross between WKY and WKHA genitors, the abundance of ventricular ANF mRNA and peptide correlated inversely with left ventricular mass, in contrast to the positive correlation observed with beta-myosin heavy chain mRNA. Finally, in the equally hypertensive SHR and WKHT strains, we found that ventricular mass was higher in SHR than in WKHT, yet ventricular ANF mRNA was less abundant in SHR than in WKHT. These results demonstrate for the first time that low ventricular ANF gene expression can be linked genetically to high cardiac mass independently of blood pressure and are consistent with a protective role for ANF against left ventricular hypertrophy. PMID- 10381899 TI - Ca2+ waves during triggered propagated contractions in intact trabeculae. Determinants of the velocity of propagation. AB - During triggered propagated contractions, Ca2+ waves travel along cardiac trabeculae with a constant velocity (Vprop) ranging from 0. 34 to 5.47 mm/s. To explore the determinants of Vprop, we studied (1) the relationship between [Ca2+]i and Vprop and (2) the effect of low concentrations of caffeine on Vprop. Trabeculae were dissected from the right ventricle of rat hearts. [Ca2+]i was measured using electrophoretically injected fura-2 and an image-intensified CCD camera. Force was measured using a silicon strain gauge, and sarcomere length was measured using laser diffraction techniques. After induction of reproducible Ca2+ waves by trains of electrical stimuli (2.5 Hz) at 21.9+/-0.2 degrees C, the number of stimuli or [Ca2+]o was varied in 9 trabeculae. In 5 trabeculae, the effects of caffeine (0.1 to 1.0 mmol/L) at [Ca2+]o of 2.2+/-0.3 mmol/L were determined. All images were recorded under stable conditions of wave propagation. The increment in [Ca2+]i during the last electrically stimulated transient (DeltaCaT) and [Ca2+]i just before onset of the Ca2+ waves (CaD) were used to estimate the Ca2+ loading of the sarcoplasmic reticulum (SR) and the myoplasm, respectively. The ratio (DeltaCaW/DeltaCaT) of the [Ca2+]i increment during the waves (DeltaCaW) to DeltaCaT was used to estimate the probability of opening of the SR-Ca2+ release channel during wave propagation. As a result of an increase of the number of stimuli or [Ca2+]o, Vprop increased in proportion to (1) DeltaCaT (r=0.82); (2) CaD (r=0.88); (3) DeltaCaW (r=0.85); and (4) DeltaCaW/DeltaCaT (r=0.74). The addition of caffeine ( 0.4). The trial was completed by 846 patients without protocol violation. Overall healing rates of reflux esophagitis were 80.4% (95% CI, 77.7-83.1) and 93.6% (95% CI, 91.8-95.2) after 4 and 8 weeks, respectively. In H. pylori-positive patients, healing rates were significantly higher after 4 (86.6% vs. 76.3%; P = 0.0005) and 8 weeks (96.4% vs. 91.8%; P < 0.004). Relief of symptoms after 4 weeks was also significantly (P < 0.05) better in H. pylori infected patients than in uninfected patients. CONCLUSIONS: Patients with reflux esophagitis and H. pylori infection respond significantly better than H. pylori negative patients to the PPI pantoprazole. PMID- 10381905 TI - Effects of very low dose daily, long-term aspirin therapy on gastric, duodenal, and rectal prostaglandin levels and on mucosal injury in healthy humans. AB - BACKGROUND & AIMS: The safety of low-dose daily aspirin therapy in the gastrointestinal tract is uncertain. Our objectives were to evaluate the long term effects of very low daily aspirin doses in the gastrointestinal tract and effects on platelet-derived serum thromboxane levels in volunteers. METHODS: Subjects were randomized to receive 10 mg (n = 8), 81 mg (n = 11), or 325 mg (n = 10) aspirin daily for 3 months. Before administration of aspirin, all subjects underwent gastroduodenoscopy, and most underwent proctoscopy for assessment of mucosal injury and prostaglandin content. After 1.5 and 3 months, subjects again underwent gastroduodenoscopy and, at 3 months, another proctoscopy. RESULTS: Each aspirin dose (even 10 mg) significantly reduced gastric mucosal prostaglandin levels, to approximately 40% of the baseline value. All three doses also induced significant gastric injury, and 325 mg caused duodenal injury. Three subjects developed gastric ulcers, 1 while taking 10 mg/day of aspirin. Furthermore, aspirin at 81 mg/day and 325 mg/day (but not 10 mg/day) significantly reduced duodenal mucosal prostaglandin levels to approximately 40% of the baseline value. Only 325 mg of aspirin per day significantly reduced rectal mucosal prostaglandin levels to approximately 60% of the baseline value. Serum thromboxane levels were inhibited 62%, 90%, and 98% with 10, 81, and 325 mg of aspirin. CONCLUSIONS: The findings explain aspirin's predominant gastric toxicity and question the safety of even 10 mg of aspirin daily. PMID- 10381906 TI - Association of HLA-DR and -DQ alleles with idiopathic achalasia. AB - BACKGROUND & AIMS: Idiopathic achalasia is a motility disorder of the esophagus characterized by incomplete relaxation of the lower esophageal sphincter and a loss of normal peristaltic activity in the body of the esophagus. The loss of inhibitory neurons in the distal esophagus, as well as abnormalities in the vagus nerve, dorsal motor nucleus of the vagus nerve, and autonomic nervous system, have been described in achalasia. Although the underlying cause of idiopathic achalasia is unknown, the diffuse neuronal effects found suggest a possible viral or neurodegenerative mechanism. By use of serological methods, a significant association between the HLA-DQ1 phenotype and idiopathic achalasia has been found, suggesting a possible immunogenetic mechanism. To further define immunogenetics in the pathogenesis of idiopathic achalasia, we performed tissue typing in patients with achalasia to determine their specific HLA phenotypes. METHODS: We prospectively studied 32 patients (23 white and 9 black) with idiopathic achalasia. Peripheral blood was collected, and HLA-DR and -DQ typing by polymerase chain reaction with sequence-specific primers was performed. Results were compared with those from 268 racially matched local controls. RESULTS: Idiopathic achalasia and the broad HLA-DQ1 allele were not significantly associated in either population, although a trend was found in white subjects (odds ratio [OR], 2.16; chi2 = 5.36, P corrected [Pc] = 0.0824). Further subtyping in white subjects revealed a significant association between idiopathic achalasia and the DQB1*0602 allele (OR, 3.10; chi2 = 7.32, Pc = 0.0408). A strong trend was also found with the DRB1*15 allele (OR, 2.83; chi2 = 8.11, Pc = 0.0572). In the black population, there was no association between idiopathic achalasia and DQB1*0602 or DRB1*15, but a trend was found with DRB1*12 (OR, 6. 19; chi2 = 5.19, P = 0.0227 uncorrected, Pc = 0.295). CONCLUSIONS: Idiopathic achalasia is associated with HLA alleles in a race-specific manner. These results support an immunogenetic mechanism in the pathogenesis of idiopathic achalasia. PMID- 10381907 TI - The cortical topography of human anorectal musculature. AB - BACKGROUND & AIMS: The muscles of the anorectum are important in the volitional control of continence, yet virtually no information exists on their cortical representation in humans. METHODS: Topographic cortical mapping of both cerebral hemispheres was performed in 9 healthy subjects by applying suprathreshold transcranial magnetic stimulation to individual points on a scalp grid centered over the vertex and then recording the electromyographic responses from the external anal sphincter, rectum, and tibialis anterior muscles. RESULTS: Cortically evoked anal and rectal response latencies were similar (20.2 +/- 1.7 and 19.8 +/- 1.5 milliseconds, respectively) and were shorter than those from the anterior tibialis muscle (right, 29.7 +/- 2.3 milliseconds; left, 29.9 +/- 1.8 milliseconds; P < 0.0005). Cortical mapping showed that the anal responses were bilaterally represented on the superior motor cortex (Brodmann area 4) of both cerebral hemispheres; a similar topography was found for the rectal responses. By comparison, the tibialis responses showed predominantly contralateral medial motor cortex representation. Subtle but consistent differences in the degree of bilateral hemispheric representation were also apparent both between and within individuals for the anal responses and to a lesser extent for the rectal responses. CONCLUSIONS: The anorectal musculature has bilateral motor cortex representation with similar topography, but there is intersubject variation in the degree of symmetry. PMID- 10381909 TI - Clinical course and costs of care for Crohn's disease: Markov model analysis of a population-based cohort. AB - BACKGROUND & AIMS: Crohn's disease results in substantial morbidity and high use of health services. The aim of this study was to describe the lifetime clinical course and costs of Crohn's disease in a 24-year population-based inception cohort of patients with Crohn's disease in Olmsted County, Minnesota. METHODS: Disease states were defined by medical and surgical treatment. A Markov model analysis calculated time in each disease state and present value of excess lifetime costs in comparison with an age- and sex-matched cohort. RESULTS: For a representative patient, projected lifetime costs were $39,906 per patient using median charges and $125,404 using mean charges. There were 29.1 years (63% of total) without medications. There were 12.7 years (27%) on aminosalicylate therapy, generating $11,467 (29%) in charges, and 3.2 years (7%) on corticosteroid or immunosuppressive therapy, generating $5147 (13%) in charges. Surgery generated $17,526 (44%) in charges. CONCLUSIONS: Most of the clinical course is spent in remission, either medical or surgical. Aminosalicylate therapy accounts for 29% of the costs of care. Surgery has the highest charges but the longest remissions. Treatment strategies that induce remission in mild disease and maintain remission with lower-cost maintenance therapy will have the largest effect on patient outcomes and costs. PMID- 10381908 TI - Recombinant Norwalk virus-like particles given orally to volunteers: phase I study. AB - BACKGROUND & AIMS: Norwalk virus (NV) is a major cause of epidemic gastroenteritis. The NV capsid is composed of a single protein that forms recombinant (rNV) virus-like particles (VLPs). In mice, these VLPs are immunogenic when administered orally without adjuvant, and they elicit serum immunoglobulin (Ig) G and intestinal IgA responses. The aim of this study was to evaluate the safety and immunogenicity of rNV VLPs in healthy volunteers. METHODS: Twenty antibody-positive adults were orally administered rNV VLPs in sterile Milli-Q water on days 1 and 21. Vaccine safety and serum rNV-specific total and subclass IgG and IgA antibody responses were monitored. The immune response induced by the VLPs was compared with the response elicited by replicating virus. RESULTS: No side effects were observed or reported by the volunteers. Serum IgG responses to rNV VLPs were dose-dependent, and all vaccinees given 250 microgram of rNV VLPs responded with >/=4-fold increases in serum IgG titers. Most of the volunteers (83%; 15 of 18) responded after the first rNV VLP dose and showed no increase in serum IgG titer after the second dose. CONCLUSIONS: Orally administered rNV VLPs are safe and immunogenic in healthy adults when administered without adjuvant and are useful to test the mucosal delivery of immunogens. PMID- 10381910 TI - Preliminary evaluation of safety and activity of recombinant human interleukin 11 in patients with active Crohn's disease. AB - BACKGROUND & AIMS: Recombinant human interleukin 11 (rhIL-11) is a cytokine with thrombocytopoietic activity and anti-inflammatory and mucosal protective effects. The objectives of this study were to investigate the safety and tolerability of rhIL-11 in patients with Crohn's disease and to explore the effects of dose and schedule on platelet count and Crohn's disease activity. METHODS: A multicenter, double-masked, placebo-controlled, dose-escalation study of 76 patients with active Crohn's disease was performed. Patients were randomized to receive subcutaneous placebo or rhIL-11 at doses of 5, 16, or 40 microgram. kg-1. wk-1 given 2 or 5 times weekly for 3 weeks. Clinical and laboratory safety data were recorded, and disease activity was measured at each visit. RESULTS: Subcutaneous injection of rhIL-11 generally was well tolerated. Significantly greater increases in platelet counts were found among patients receiving rhIL-11 40 microgram. kg-1. wk-1 as 2 or 5 weekly doses and 16 microgram. kg-1. week-1 as 5 weekly doses compared with patients receiving placebo (P < 0.05). Patients receiving 16 microgram. kg-1. wk-1 had the highest clinical response rates, with a response seen in 42% of patients (5/12) receiving 5 weekly doses and 33% of patients (4/12) receiving 2 weekly doses, compared with 7% of patients (1/15) receiving placebo. CONCLUSIONS: Short-term treatment with rhIL-11 is well tolerated in patients with active Crohn's disease. The thrombocytopoietic effect of rhIL-11 seems to be both dose and schedule dependent and may be minimized with retained clinical benefit in Crohn's disease at 16 microgram. kg-1. wk-1 given in 2 equal doses. PMID- 10381911 TI - Epithelial permeability to proteins in the noninflamed ileum of Crohn's disease? AB - BACKGROUND & AIMS: Crohn's disease (CD) is associated with a disturbed intestinal barrier. Permeability studies have focused on inert molecules, but little is known about transepithelial transport of macromolecules with antigenic potential in humans. The aim of this study was to quantify permeation and to characterize passage routes for macromolecules in ileal mucosa in CD. METHODS: Noninflamed and inflamed ileal mucosa specimens from patients with CD (n = 12) and ileal specimens from patients with colon cancer (n = 7) were studied regarding transmucosal permeation of ovalbumin, dextran (mol wt, 40,000), and 51Cr-EDTA for 90 minutes in vitro in Ussing chambers. Transepithelial passage routes for fluorescent ovalbumin and dextran 40,000 were investigated by confocal microscopy. RESULTS: Noninflamed ileum from CD patients showed increased permeation of ovalbumin compared with ileum from colon cancer patients (P < 0.05). Dextran permeation was equal in the three groups, whereas 51Cr-EDTA permeability was increased in inflamed ileum. Ovalbumin passed both transcellularly and paracellularly, but dextran followed a strictly paracellular route. Both markers were subsequently endocytosed by cells of the lamina propria. CONCLUSIONS: Noninflamed ileal mucosa from patients with CD shows increased epithelial permeability to ovalbumin, probably by augmented transcytosis. This increase in antigen load to the lamina propria could be an initiating pathogenic event in CD. PMID- 10381912 TI - Inflammatory alterations in mesenteric adipose tissue in Crohn's disease. AB - BACKGROUND & AIMS: Abnormalities of fat in the mesentery including adipose tissue hypertrophy and fat wrapping have been long recognized on surgical specimens as characteristic features of Crohn's disease. However, the importance, origin, and significance of the mesenteric fat hypertrophy in this chronic inflammatory disease are unknown. Peroxisome proliferator-activated receptor gamma (PPARgamma) is a crucial factor involved in the homeostasis of adipose tissue, a major source of biologically active mediators. METHODS: Intra-abdominal fat accumulation was quantified using a magnetic resonance imaging method in patients with Crohn's disease and controls. PPARgamma and inflammatory cytokines synthesized by mesenteric adipose tissues were assessed by quantitative polymerase chain reaction, in situ hybridization, and immunohistochemistry. RESULTS: In vivo, patients with Crohn's disease have an important accumulation of intra-abdominal fat. This mesenteric obesity, present from the onset of the disease, is associated with overexpression of PPARgamma and tumor necrosis factor (TNF) alpha, synthesized, at least in part, by adipocytes. CONCLUSIONS: These results suggest that confined increased PPARgamma mesenteric concentrations could lead to the mesenteric fat hypertrophy, which could actively participate through the synthesis of TNF-alpha in the inflammatory response. PMID- 10381913 TI - Isolation of a Helicobacter pylori protein, FldA, associated with mucosa associated lymphoid tissue lymphoma of the stomach. AB - BACKGROUND & AIMS: The growth of gastric mucosa-associated lymphoid tissue lymphoma (MALToma) seems to depend on the stimulation of Helicobacter pylori. We attempted to identify specific antigen(s) from H. pylori strains associated with MALToma. METHODS: Membranous and secreted proteins of H. pylori were compared on sodium dodecyl sulfate-polyacrylamide gel electrophoresis by Western blot using sera from patients with MALToma. RESULTS: A 19-kilodalton protein was seen in all strains isolated from patients with MALToma but uncommonly in other strains. The protein was purified and sequenced. Amino acid sequence comparison showed it was an FldA homologue, a putative flavodoxin protein. DNA sequencing in 26 strains revealed that a nucleotide G insertion at position 481 of the fldA gene was more frequently observed in strains associated with MALToma than other strains (9/9 vs. 6/17; P = 0.002). The mutation caused a short truncation. A recombinant protein with this truncation was expressed and tested. Sera of 12 (70.6%) of 17 patients with MALToma were positive for the antibody to the recombinant protein, and 7 (16.7%) of 42 control patients were positive (12/17 vs. 7/42; P < 0.0001). CONCLUSIONS: Truncated FldA of H. pylori is associated with gastric MALToma. It may be involved in the pathogenesis of gastric MALToma. Antibody to this antigen could be used as a serological marker of the disease. PMID- 10381914 TI - Effects of a chargrilled meat diet on expression of CYP3A, CYP1A, and P glycoprotein levels in healthy volunteers. AB - BACKGROUND & AIMS: Carcinogenic heterocyclic amines and polycyclic aromatic hydrocarbons present in chargrilled meat are substrates for inducible CYP1A and CYP3A enzymes and for P-glycoprotein. We examined whether consumption of a chargrilled meat diet results in induction of these proteins. METHODS: Ten healthy adults were fed a diet enriched with chargrilled meat for 7 days. Duodenal biopsy specimens were obtained on days 1, 5, and 12 and analyzed for CYP1A, CYP3A, and P-glycoprotein messenger RNA (mRNA) and protein. On days 5 and 12, hepatic CYP3A4 and CYP1A2 activities were measured and colon biopsies were performed. The levels of polycyclic aromatic hydrocarbon DNA adducts in peripheral blood mononuclear cells were measured on days 1, 4, 11, and 26. RESULTS: There was no detectable induction of CYP3A4, CYP3A5, or P-glycoprotein mRNAs or protein in small intestine or colon and no induction of hepatic CYP3A4 enzyme activity. In contrast, the chargrilled meat diet resulted in unequivocal induction of CYP1A enzymes in the liver and small intestine of each subject. There was an inverse correlation between the level of peripheral polycyclic aromatic hydrocarbon DNA adducts measured on day 11 and both liver CYP1A2 activity (P = 0.027) and enterocyte CYP1A1 protein concentration (P = 0.046). CONCLUSIONS: Ingestion of chargrilled meat results in induction of CYP1A enzymes but not CYP3A4 or P-glycoprotein. This observation, combined with the correlation between adduct levels and CYP1A expression, supports an adaptive role for CYP1A but not CYP3A4 or P-glycoprotein. PMID- 10381915 TI - Secretion of the intestinotropic hormone glucagon-like peptide 2 is differentially regulated by nutrients in humans. AB - BACKGROUND & AIMS: Glucagon-like peptide 2(1-33) (GLP-2(1-33)), an intestinally derived hormone, stimulates growth in rodent small and large bowel. To explore the physiology of GLP-2(1-33) secretion, we measured plasma GLP-2 levels in 6 healthy male volunteers, before and after test meals. METHODS: Blood samples were collected over 24 hours with the subjects consuming a normal, solid mixed diet (2500 kcal) and for 4 hours after liquid test meals (400 kcal/300 mL) composed of carbohydrate, fat, or protein. All studies commenced at 9 AM. Plasma was extracted and analyzed in radioimmunoassays for N-terminal immunoreactive GLP-2 (N-IR-GLP-2; measures bioactive GLP-2(1-33)) as well as total IR-GLP-2 (T-IR-GLP 2), which includes GLP-2(1-33), GLP-2(3-33) (an inactive degradation product of GLP-2(1-33)), and the pancreatic major proglucagon fragment (an inactive precursor that contains GLP-2). Basal and nutrient-stimulated plasma samples were also analyzed by high-performance liquid chromatography to determine the levels of GLP-2(1-33) and GLP-2(3-33). RESULTS: N-IR-GLP-2 levels were increased 2.0 +/- 0.2- to 2.8 +/- 0.5-fold 40 minutes after each mixed meal (P < 0.05-0.01) and returned to basal overnight, whereas T-IR-GLP-2 levels were increased 1.3 +/- 0.1 fold 40 minutes after breakfast only (P < 0.05). After ingestion of carbohydrate or fat alone, plasma N-IR-GLP-2 concentrations increased by 5.6 +/- 2.0- and 2.7 +/- 0.6-fold within 1 hour (P < 0.05). High-performance liquid chromatography analysis showed a relative increase in the levels of GLP-2(1-33) compared with GLP-2(3-33) (P < 0.05). Ingestion of the protein meal did not alter N-IR-GLP-2 levels, whereas T-IR-GLP-2 was increased by fat and protein (by 1.7 +/- 0. 2-fold for each, P < 0.01) but not by carbohydrate. CONCLUSIONS: These results show that secretion of GLP-2(1-33) from the intestine is regulated in a nutrient-dependent manner in normal humans. PMID- 10381916 TI - Protein kinase C-dependent activation of NF-kappaB in enterocytes is independent of IkappaB degradation. AB - BACKGROUND & AIMS: Nuclear translocation of the NF-kappaB family of transcription factors is a proximal step in the signal transduction of a pleiotropic group of proinflammatory genes. Activation of RelA is under the negative control of IkappaB, a family of proteins degraded in response to immunologic and oxidant stimuli. The aim of this study was to examine this mechanism of NF-kappaB activation in intestinal epithelial cells. METHODS: DLD-1 cell monolayers stimulated by interleukin (IL)-1beta or phorbol myristate acetate (PMA) were assayed for the nuclear translocation of NF-kappaB and immunoreactivity of various IkappaB isoforms that regulate NF-kappaB1/RelA activation. RESULTS: NF kappaB activation triggered by PMA was not associated with the disappearance of immunoreactive IkappaBalpha, IkappaBbeta, IkappaBgamma, or IkappaBepsilon or with the dissociation of intact IkappaB from RelA. NF-kappaB activation induced by PMA was blocked by the protein kinase C inhibitor staurosporine but not by the proteasomal inhibitor N-acetyl-leucine leucine norleucinal (ALLN). In contrast, IL-1beta-induced NF-kappaB activation was associated with the disappearance of IkappaBalpha and was inhibited by ALLN but not staurosporine. CONCLUSIONS: Our data imply the existence of a novel pathway of NF-kappaB activation mediated by protein kinase C that does not require proteosomal degradation or the loss of IkappaB. PMID- 10381917 TI - Heat-shock protein 72 protects against oxidant-induced injury of barrier function of human colonic epithelial Caco2/bbe cells. AB - BACKGROUND & AIMS: Barrier function of the inflamed intestinal mucosa can be compromised by reactive oxygen metabolites that increase mucosal permeability and disrupt the actin cytoskeleton, the integrity of which is important for maintaining tight epithelial junctions. Because heat-shock protein 72 (hsp72) protects intestinal epithelial cells against injury, we determined whether resistance of Caco2/bbe (C2) intestinal monolayer barrier function was related to their high endogenous hsp72 expression. METHODS: hsp72 anti-sense (C2/AS) and vector-only transfected C2 (C2/CEP4) clones, lines that exhibit low and high hsp72 expression, respectively, were studied. Permeability was assessed by measuring electrical resistance and mannitol fluxes and actin organization by confocal fluorescein isothiocyanate-phalloidin analysis. RESULTS: Basal transepithelial electrical resistance (TER) and mannitol fluxes were not significantly different between groups. However, the oxidant monochloramine rapidly decreased TER and increased mannitol permeability of C2/AS monolayers compared with C2/CEP4 (50% effective doses at 30 minutes were 0.53 +/- 0.11 and 2.06 +/- 0.34 mmol/L, respectively). Associated with these changes, decreased cell viability, dissociation and aggregation of perijunctional and stress actin filaments, loss of cell height, and increased intercellular separation were observed only in C2/AS cells treated with monochloramine. CONCLUSIONS: hsp72 protects intestinal epithelial barrier function against oxidant-induced stress, in part, by protecting the integrity of the actin cytoskeleton. PMID- 10381918 TI - Mismatch repair proficiency and in vitro response to 5-fluorouracil. AB - BACKGROUND & AIMS: The DNA mismatch repair (MMR) system recognizes certain DNA adducts caused by alkylation damage in addition to its role in recognizing and directing repair of interstrand nucleotide mismatches and slippage mistakes at microsatellite sequences. Because defects in the MMR system can confer tolerance to acquired DNA damage and, by inference, the toxic effects of certain chemotherapeutic agents, we investigated the effect of 5-fluorouracil (5-FU) on colon cancer cell lines. METHODS: We determined growth selection by cell enrichment assay and cloning efficiency after treatment with 5 micromol/L 5-FU, assayed nucleic 3H-5-FU incorporation, and analyzed the cell cycle by flow cytometry. RESULTS: 5-FU treatment provided a growth advantage for MMR-deficient cell lines, indicating a relative degree of tolerance to 5-FU by the MMR deficient cell lines. Enhanced survival was statistically significant after 5 days of growth, and a 28-fold reduction in survival was noted in the MMR proficient cells by clonagenic assays after 10 days of growth. Differences in nucleotide uptake of 5-FU did not account for the observed growth differences, and specific cell cycle checkpoint arrest was not detected. CONCLUSIONS: Intact DNA MMR seems to recognize 5-FU incorporated into DNA but may do so in a different manner than other types of alkylation damage. Defective DNA MMR might be one mechanism for tumor resistance to 5-FU. PMID- 10381919 TI - The glucose-6 phosphatase gene is expressed in human and rat small intestine: regulation of expression in fasted and diabetic rats. AB - BACKGROUND & AIMS: Glucose-6 phosphatase (Glc6Pase) is the last enzyme of gluconeogenesis and glycogenolysis, previously assumed to be expressed in the liver and kidney only, conferring on both tissues the capacity to produce endogenous glucose in blood. METHODS: Using Northern blotting and reverse transcription polymerase chain reaction and a highly specific Glc6Pase assay, we studied expression of the Glc6Pase gene in human and in rat tissues (fasted and diabetic). RESULTS: The Glc6Pase gene is expressed in the duodenum and jejunum in normal fed rats and in the duodenum, jejunum, and ileum in humans. The Glc6Pase messenger RNA (mRNA) abundance was increased eightfold and sixfold in the duodenum and jejunum of streptozotocin diabetic rats. It was normalized in both tissues after 10 hours of insulin treatment. Glc6Pase activity was increased by 300% in the duodenum and jejunum in diabetic rats compared with normal rats. The Glc6Pase mRNA abundances and enzymatic activities were increased in a similar manner in both tissues in 48-hour-fasted rats. Normalization of mRNA abundance was achieved after refeeding for 7 hours. In addition, Glc6Pase mRNA and activity were also expressed in the ileum during fasting in rats. CONCLUSIONS: These data show that the small intestine has the ability to release endogenous glucose and strongly suggest that its contribution to systemic glucose production might be increased in situations of insulinopenia (type 1 diabetes) and insulin resistance (type 2 diabetes and others). PMID- 10381920 TI - Blockade of kit signaling induces transdifferentiation of interstitial cells of cajal to a smooth muscle phenotype. AB - BACKGROUND & AIMS: Interstitial cells of Cajal (ICC) serve as pacemaker cells and mediators of neurotransmission from the enteric nervous system to gastrointestinal muscles. ICC develop from mesenchymal cells that express c-Kit, and signaling via Kit receptors is necessary for normal development of ICC. We studied the fate of functionally developed ICC after blockade of Kit receptors to determine whether ICC undergo cell death or whether the phenotype of the cells is modified. The fate of undeveloped ICC was also investigated. METHODS: Neutralizing, anti-Kit monoclonal antibody (ACK2) was administered to mice for 8 days after birth. ICC in the small intestine were examined by immunohistochemistry and electron microscopy. Occurrence of apoptosis was also assayed. RESULTS: When Kit receptors were blocked, ICC nearly disappeared from the small intestine. Apoptosis was not detected in regions where ICC are normally distributed. Remaining Kit-immunopositive cells in the pacemaker region of the small intestine developed ultrastructural features similar to smooth muscle cells, including prominent filament bundles and expression of the muscle-specific intermediate filament protein, desmin, and smooth muscle myosin. ICC of the deep muscular plexus normally develop after birth in the mouse. Precursors of these cells remained in an undifferentiated state when Kit was blocked. CONCLUSIONS: These data, along with previous studies showing that ICC in the pacemaker region of the small intestine and longitudinal muscle cells develop from the same Kit immunopositive precursor cells, suggest inherent plasticity between the ICC and smooth muscle cells that is regulated by Kit-dependent cell signaling. PMID- 10381921 TI - Outcome of liver transplantation in hepatitis C virus-infected patients who received hepatitis C virus-infected grafts. AB - BACKGROUND & AIMS: The present organ shortage has brought into question the suitability of hepatitis C virus (HCV)-positive grafts. This study reviewed the outcome of such transplantations in our institution. METHODS: Twenty-three HCV positive patients who underwent orthotopic liver transplantation (OLT) for end stage liver disease with HCV-positive grafts in 1992-1995 were studied. Only patients who survived more than 30 days were included in the analysis. Control group included 169 patients who underwent transplantation for HCV-related cirrhosis and received HCV-negative organs. RESULTS: Patients who received HCV infected organs had a cumulative survival rate of 89% and 72% at 1 and 5 years, respectively, vs. 88% and 73% for the control group (NS). There was no difference in graft survival, incidence of cirrhosis, mean hepatitis activity index score, fibrosis, or mean activity of serum transaminases. There was a trend toward lower incidence of recurrent hepatitis C in the study group compared with control (21% vs. 23% at 1 year and 47% vs. 64% at 5 years; NS). Patients in whom the donor strain became predominant after transplantation had significantly longer disease free survival than patients who retained their own HCV strain (P < 0.003). CONCLUSIONS: HCV-infected livers transplanted into HCV-infected recipients do not appear to convey a worse outcome in the initial years after OLT than HCV-negative grafts. PMID- 10381922 TI - p53 mutations in british patients with hepatocellular carcinoma: clustering in genetic hemochromatosis. AB - BACKGROUND & AIMS: Environmental factors are important in the etiology of hepatocellular carcinoma (HCC). Aflatoxin B1 causes a specific point mutation in the p53 tumor-suppressor gene in exposed individuals. In Western populations, mutations of this gene seem to be less frequent. We have investigated the role of p53 mutations in tumorigenesis in British patients with HCC. The aim of this study was to determine the frequency and mutational spectrum of the p53 gene in HCCs from British patients. METHODS: DNA from 170 HCCs, of well-defined etiology, in British patients was analyzed by single-stranded conformational polymorphism using the polymerase chain reaction technique. Mutations were then characterized by direct sequencing. RESULTS: Twenty-nine percent of tumors had p53 mutations. Ten of 14 (71%) hemochromatotic cancers had mutations within the p53 gene, and clustering of these mutations at codon 220 (A-G) was found in 5 cases; 3 others had T-A mutations. No clustering was found in HCCs with other etiologies. CONCLUSIONS: p53 mutations are more common than was thought in Northern European HCCs. This is the first demonstration of p53 mutational clustering in HCCs from hemochromatotic subjects. PMID- 10381923 TI - Changing pattern of chronic hepatitis D in Southern Europe. AB - BACKGROUND & AIMS: The aim of this study was to assess changes in the clinical pattern of hepatitis D virus (HDV) infection in Italy, brought about by improved control of hepatitis B and D viruses, and to establish the natural history of chronic hepatitis D. METHODS: Histological diagnosis and clinical features of 122 patients with HDV recruited from 1987 to 1996 in three Italian tertiary referral centers (Torino, northern Italy; San Giovanni Rotondo and Castellana Grotte, southern Italy) were compared with those of 162 patients collected in the same centers in the previous decade. Patients from both groups with at least 6 months of follow-up were included in a new subgroup to assess the natural history of the disease. RESULTS: Among 162 patients referred from 1977 to 1986, 9 (6%) had mild hepatitis at histology vs. 9 (8%) of 122 patients referred in the second decade; 105 (65%) vs. 21 (17%) had severe hepatitis; 46 (28%) vs. 38 (31%) had histological asymptomatic cirrhosis; and 2 (1%) vs. 54 (44%) had clinically overt cirrhosis. For 159 patients (121 men and 38 women; mean age, 34 +/- 11), a follow up of more than 6 months was documented, and they were included in the natural history subgroup. After 78 +/- 59 months of follow-up, 112 (70%) survived free of liver transplantation: 9 underwent transplantation, 32 died of liver failure, and 6 of acquired immunodeficiency syndrome. Estimated 5- and 10-year probability of survival free of orthotopic liver transplantation was 100% and 100% for patients with mild hepatitis, 90% and 90% for severe hepatitis, 81% and 58% for histological asymptomatic cirrhosis, and 49% and 40% for clinical cirrhosis (P < 0.01), respectively. CONCLUSIONS: Occurrence of fresh and severe forms of hepatitis D has diminished greatly in Italy. Contemporary patients represent cohorts infected years ago who survived the immediate medical impact of hepatitis D. The disease has been asymptomatic and nonprogressive in a minority; in the majority, it rapidly advanced to cirrhosis but thereafter subsided with stable clinical conditions for more than a decade. PMID- 10381924 TI - Assessment of biliary bicarbonate secretion in humans by positron emission tomography. AB - BACKGROUND & AIMS: Positron emission tomography (PET) allows imaging and quantitative analysis of organ functions in basal and stimulated conditions. We have applied this method to the study of biliary bicarbonate secretion in humans. METHODS: PET was performed in 5 healthy subjects and 13 patients with hepatobiliary disorders after intravenous injection of NaH11CO3. In each case the study was performed in basal conditions and after secretin stimulation. Positron emission from the hepatic area was scanned, and normalized uptake values for parenchymal and hilar regions were estimated. RESULTS: In healthy individuals, the injection of NaH11CO3 resulted in a peak uptake of the label in parenchymal and hilar regions 2-3 minutes after the injection. In both normal and cirrhotic subjects, secretin administration increased bicarbonate uptake in the parenchymal region, followed by accumulation of the label in the perihilar area. Normal basal uptake with absent response to secretin was registered in extrahepatic biliary obstruction and in untreated primary biliary cirrhosis (PBC). The secretin response was present in patients with PBC undergoing treatment with ursodeoxycholic acid. CONCLUSIONS: PET allows investigation of biliary bicarbonate secretion in humans. An impaired response to secretin was observed in cholestatic conditions. Preliminary data suggest that ursodeoxycholic acid might improve the response to secretin in PBC. PMID- 10381925 TI - Nitric oxide increases hepatic arterial blood flow in rats with carbon tetrachloride-induced acute hepatic injury. AB - BACKGROUND & AIMS: Little is known about the changes in hepatic blood flow associated with acute hepatic injury. The aim of this study was to investigate the effect of nitric oxide (NO) on hepatic blood flow and the severity of hepatic injury in rats with carbon tetrachloride (CCl4)-induced acute hepatic injury. METHODS: Rats were pretreated with CCl4 to induce acute hepatic injury. Hepatic blood flow was measured using a radioactive microsphere method. The role of NO in the regulation of hepatic blood flow and the severity of hepatic injury was investigated by administering NG-nitro-L-arginine (L-NNA) and aminoguanidine (AG). Plasma nitrite/nitrate levels, hepatic NO synthase (NOS) activity, and expression of hepatic NOS messenger RNA (mRNA) were measured, and histological examinations were performed. RESULTS: Hepatic arterial and portal venous blood flow was increased significantly by CCl4, without any change in mean arterial pressure or cardiac output. L-NNA and AG dose-dependently decreased hepatic arterial blood flow, with the highest dose resulting in complete blockade of hepatic arterial dilation, but failed to change portal venous blood flow. Histologically, administration of AG aggravated the hepatic injury in CCl4 treated rats. Plasma nitrite/nitrate levels and hepatic NOS activity were increased significantly by CCl4 treatment. Inducible NOS mRNA was detected in CCl4-treated rats but not in the controls. CONCLUSIONS: The results of this study suggest that the increased hepatic arterial blood flow in CCl4-induced acute hepatic injury is mediated by excessive NO production and up-regulated by inducible NOS, which plays a role in reducing hepatic injury. PMID- 10381926 TI - An alcoholic binge causes massive degradation of hepatic mitochondrial DNA in mice. AB - BACKGROUND & AIMS: Ethanol causes oxidative stress in the hepatic mitochondria of experimental animals and mitochondrial DNA deletions in alcoholics. We postulated that ethanol intoxication may cause mitochondrial DNA strand breaks. METHODS: Effects of an intragastric dose of ethanol (5 g/kg) on hepatic mitochondrial DNA levels, structure, and synthesis were determined by slot blot hybridization, Southern blot hybridization, and in vivo [3H]thymidine incorporation, respectively. RESULTS: Two hours after ethanol administration, ethane exhalation (an index of lipid peroxidation) increased by 133%, although hepatic lipids were unchanged. Mitochondrial DNA was depleted by 51%. Its supercoiled form disappeared, whereas linearized forms increased. Long polymerase chain reaction evidenced lesions blocking polymerase progress on the mitochondrial genome. Mitochondrial transcripts decreased. Subsequently, [3H]thymidine incorporation into mitochondrial DNA increased, and mitochondrial DNA levels were restored. In contrast, nuclear DNA was not fragmented and its [3H]thymidine incorporation was unchanged. Liver ultrastructure only showed inconstant mitochondrial lesions. Ethanol-induced mitochondrial DNA depletion was prevented by 4-methylpyrazole, an inhibitor of ethanol metabolism, and attenuated by melatonin, an antioxidant. CONCLUSIONS: After an alcoholic binge, ethanol metabolism causes oxidative stress and hepatic mitochondrial DNA degradation in mice. DNA strand breaks may be involved in the development of mitochondrial DNA deletions in alcoholics. PMID- 10381928 TI - Glutathione protects the rat liver against reperfusion injury after hypothermic preservation. AB - BACKGROUND & AIMS: The extracellular generation of reactive oxygen species (ROS) by Kupffer cells contributes to reperfusion injury of the liver allograft. The endogenous antioxidant glutathione (GSH) can detoxify these ROS; however, this effect might be limited by the low extracellular concentration of GSH. We therefore investigated whether an increase of extracellular GSH protects the liver against reperfusion injury after cold preservation. METHODS: Livers of male Sprague-Dawley rats subjected to 24 hours of cold ischemia in University of Wisconsin solution (4 degrees C) were reperfused for 2 hours in the absence (controls) or presence of 0.5, 1, 2, or 4 mmol/L GSH (n = 4-6 each). RESULTS: Two hours after starting reperfusion of control livers, the sinusoidal release of lactate dehydrogenase and purine nucleoside phosphorylase increased to 247 +/- 96 and 27 +/- 13 mU. min(-1). g liver(-1), respectively, but only to 76 +/- 43 and 10 +/- 4 mU. min(-1). g liver(-1) in the presence of 4 mmol/L GSH. This cytoprotective effect was confirmed histologically by a marked reduction of trypan blue staining of hepatocytes. Compared with control livers, postischemic bile flow was significantly enhanced by GSH (0.15 +/- 0.02 vs. 0.41 +/- 0.11 microL. min(-1). g liver(-1)), indicating improved liver function. During reperfusion of control livers, intracellular GSH content declined from 4.5 +/- 0.3 to 2.3 +/- 0.1 micromol/g liver, but only to 3.8 +/- 0.4 micromol/g liver in the presence of 4 mmol/L GSH. Reperfusion of untreated livers was accompanied by a prolonged increase of portal pressure to maximally 12.5 +/- 1.9 cm H2O, which was significantly attenuated by 4 mmol/L GSH (7.2 +/- 1.4 cm H2O). Similar cytoprotective and hemodynamic effects were observed with 2 mmol/L GSH, but not with 0.5 and 1 mmol/L GSH. CONCLUSIONS: Treatment of cold-preserved livers with GSH upon reperfusion prevents damage of hepatocytes, deterioration of the hepatic circulation, and loss of intracellular GSH. In view of these protective effects and its low toxicity in humans, GSH should be considered a candidate drug for prevention of ROS-related reperfusion injury of the liver allograft. PMID- 10381927 TI - Cholinergic system modulates growth, apoptosis, and secretion of cholangiocytes from bile duct-ligated rats. AB - BACKGROUND & AIMS: To investigate the role of the cholinergic system in regulation of cholangiocyte functions, we evaluated the effects of vagotomy on cholangiocyte proliferation and secretion in rats that underwent bile duct ligation (BDL rats). METHODS: After bile duct ligation (BDL), the vagus nerve was resected; 7 days later, expression of M3 acetylcholine receptor was evaluated. Cholangiocyte proliferation was assessed by morphometry and measurement of DNA synthesis. Apoptosis was evaluated by light microscopy and annexin-V staining. Ductal secretion was evaluated by measurement of secretin-induced choleresis, secretin receptor (SR) gene expression, and cyclic adenosine 3',5'-monophosphate (cAMP) levels. RESULTS: Vagotomy decreased the expression of M3 acetylcholine receptors in cholangiocytes. DNA synthesis and ductal mass were markedly decreased, whereas cholangiocyte apoptosis was increased by vagotomy. Vagotomy decreased ductal secretion. Forskolin treatment prevented the decrease in cAMP levels induced by vagotomy, maintained cholangiocyte proliferation, and decreased cholangiocyte apoptosis caused by vagotomy in BDL rats. Cholangiocyte secretion was also maintained by forskolin. CONCLUSIONS: Vagotomy impairs cholangiocyte proliferation and enhances apoptosis, leading to decreased ductal mass in response to BDL. Secretin-induced choleresis of BDL rats was virtually eliminated by vagotomy in association with decreased cholangiocyte cAMP levels. Maintenance of cAMP levels by forskolin administration prevents the effects of vagotomy on cholangiocyte proliferation, apoptosis, and secretion. PMID- 10381929 TI - Ischemic necrosis of bile ducts complicating Schonlein-Henoch purpura. AB - Gastrointestinal complications of Schonlein-Henoch purpura are frequent and sometimes severe. However, there seem to be no reports of liver involvement. A child is described in whom Schonlein-Henoch purpura was complicated by bile duct lesions, resulting in biliary cirrhosis and requiring liver transplantation. At surgical removal, the liver had lesions of bile ducts and of adjacent small blood vessels in the hilum, very similar to those complicating hepatic artery thrombosis after liver transplantation. These findings suggest that Schonlein Henoch purpura can be complicated by vasculitis of the peribiliary vessels resulting in ischemic necrosis of the bile ducts. Schonlein-Henoch purpura can be added to the list of causes of ischemic cholangiopathies. PMID- 10381931 TI - The role of gastric carditis in metaplasia and neoplasia at the gastroesophageal junction. AB - Adenocarcinomas at the gastroesophageal junction appear to arise from foci of intestinal metaplasia that develop either in the distal esophagus or the proximal stomach (the gastric cardia). Metaplasia is usually a consequence of chronic inflammation, and it is logical to assume that intestinal metaplasia at the gastroesophageal junction develops as a result of chronic inflammation in the epithelia that normally line the junction region. Intestinal metaplasia in the esophagus is known to be a sequela of chronic inflammation in squamous epithelium caused by gastroesophageal reflux disease, whereas intestinal metaplasia in the distal stomach is often a consequence of chronic gastritis caused by Helicobacter pylori infection. For the gastric cardia, the contributions of gastroesophageal reflux disease, H. pylori infection, and other factors to inflammation, metaplasia, and neoplasia are not clear. If physicians are to develop meaningful preventive strategies and specific therapies for tumors of the proximal stomach, a clear understanding of pathogenesis is important. Recent studies on pathogenetic factors for inflammation in cardiac epithelium (gastric carditis) have yielded contradictory results, perhaps because of fundamental differences in the techniques used by different investigators for identifying and sampling the gastric cardia. This report explores the roots of the controversy regarding the role of gastric carditis in the development of metaplasia and neoplasia at the gastroesophageal junction and suggests practical guidelines for biopsy protocols to be used in future studies that will be necessary to resolve these disputes. PMID- 10381930 TI - Relapsing ulcerative colitis associated with spinal cord stimulation. AB - Spinal cord stimulation is an increasingly popular form of pain treatment. An electrode positioned on the dorsal aspect of the spinal cord at the level of the nerve roots from the painful area stimulates the spinal cord. Current from the electrode is supplied by a pulse generator in the lower anterior abdominal wall. Spinal cord stimulation has not previously been associated with ulcerative colitis. A man with left-sided ulcerative colitis in remission experienced two successive relapses related to the use of a spinal cord stimulation system. After removal of the system, remission returned and remained. This case suggests that electrical current may influence the course of ulcerative colitis. PMID- 10381932 TI - American gastroenterological association medical position statement on treatment of patients with dysphagia caused by benign disorders of the distal esophagus. AB - This document presents the official recommendations of the American Gastroenterological Association (AGA) on treatment of patients with dysphagia caused by benign disorders of the distal espophagus. It was approved by the Clinical Practice and Practice Economics Committee on September 27, 1998, and by the AGA Governing Board on November 8, 1998. PMID- 10381933 TI - AGA technical review on treatment of patients with dysphagia caused by benign disorders of the distal esophagus. AB - This literature review and the recommendations therein were prepared for the American Gastroenterological Association Clinical Practice and Practice Economics Committee. The paper was approved by the committee on September 27, 1998. PMID- 10381934 TI - A new model system to study Norwalk virus immunity. PMID- 10381935 TI - Helicobacter pylori factors associated with disease. PMID- 10381936 TI - Nutrient regulation of intestinal growth and adaptation: role of glucagon-like peptide 2 and the enteroendocrine cell. PMID- 10381937 TI - Will transplantation of an hepatitis C-infected graft improve the outcome of liver transplantation in HCV patients? PMID- 10381938 TI - Effects of alcohol on hepatic mitochondrial function and DNA. PMID- 10381940 TI - The appendix in ulcerative colitis: a not so innocent bystander. PMID- 10381939 TI - H. pylori and nonulcer dyspepsia: not guilty as charged. PMID- 10381941 TI - In pursuit of the hidden, the occult, and the obscure. PMID- 10381942 TI - Pancreatic bicarbonate secretion: role of CFTR and the sodium-bicarbonate cotransporter. PMID- 10381943 TI - Nutrition and Women's cancer PMID- 10381944 TI - Editorial AB - As dermatologists, we have all been active in educating patients about sun awareness and sun protection. This is even more important for children, as childhood exposure to ultraviolet light is a significant risk for both melanoma and nonmelanoma skin cancers. The importance of an educational approach in appropriate sun awareness in childhood is further underscored by the recent findings by Rivers et al., in the Vancouver Moles Cohort study, presented at the 1999 American Academy of Dermatology meeting. In a placebo-controlled trial, the findings of Rivers et al. clearly demonstrated that the use of sunscreens can significantly decrease the formation of nevae in children, providing further evidence to support sun awareness education initiatives. The lead article by Gooderham and Guenther in the Basic and Clinical Sciences section evaluates the effectiveness of a particular sun awareness program, and gives valuable insights into how more effective approaches may be used in the future. In addition to ultraviolet light playing a causal role in cutaneous malignancies, it is known to induce a number of other skin problems. One particularly difficult group of disorders is the photosensitive dermatoses, including solar urticaria. Bissonnette et al. describe an innovative approach to the management of refractory solar urticaria with plasma exchange. In the Grand Rounds section, Strauss et al. review the case of an acute SLE and give an insightful discussion related to bullous eruptions in acutely ill children. The mechanism of ultraviolet-light-induced carcinogenesis involves UV-induced DNA damage. Over the past decade, it has become clear that tumour suppressor genes can regulate these processes. In the Review section, Tron et al. discuss the role of the suppressor gene p53, which is mutated or lost in nonmelanoma skin cancer. P53 is crucial in protecting keratinocytes from the harmful effects of ultraviolet radiation, and in their instructive article, these authors use gene-targeted mutant mice lacking p53 to further evaluate the role in UV-induced DNA damage. With the warm weather upon us, we are spending more time in the outdoors and, as a result, are exposed to a vast number of environmental onslaughts. These include such things as Rickettsial disease, summarized in our CME section Summary Notes. Furthermore, in a comprehensive review, Dr. Sasseville examines another outdoor threat as he delineates the wide spectrum of plant contact dermatitis. This represents an important and in-depth reference on phytodermatitis. Our specialty, and indeed all of medicine, is being dramatically altered by recent advances in our understanding of disease at a molecular level. This new understanding of disease has led to the potential of modifying gene expression through the use of gene therapy. This is particularly attractive in skin disease, where gene therapy can be delivered quite readily through the skin. This advancement is insightfully discussed in the article by Somani et al., "Gene Therapy and Dermatology," which is both valuable for the cognoscenti and noncognoscenti alike, and serves as an important reference work in this area. PMID- 10381945 TI - Sun and the skin: evaluation of a sun awareness program for elementary school students. AB - BACKGROUND: Research suggests that childhood exposure to ultraviolet radiation is a significant risk factor for the development of melanoma and nonmelanoma skin cancers. Sun awareness education programs for children can positively influence children's sun protective practices to decrease the risk of skin cancer. OBJECTIVE: The purpose of this study was to evaluate a sun awareness education program, entitled "Sun and the Skin" that had recently been developed and implemented in London, Ontario. METHOD: The study uses a pre- and posttest design to evaluate both knowledge and behaviour of Grade 4 students participating in the program at baseline, immediately after, and 1 month after the program. RESULTS: The students demonstrated a significant increase in their sun-protective practices after participation in the "Sun and the Skin" program. There was a significant improvement in the students' level of knowledge after the program. Improvement in both behaviour and knowledge were maintained weeks after completion of the program. Minor differences in knowledge due to demographic characteristics were detected after the program. CONCLUSIONS: A sun awareness education program for Grade 4 students can improve both their knowledge and behaviour over time. This format should be used in conjunction with changes in school policy in order to make a significant longterm impact. PMID- 10381946 TI - Treatment of refractory solar urticaria with plasma exchange. AB - BACKGROUND: Solar urticaria is a photodermatosis that can be very disabling for patients who are highly sensitive to light and can also be very resistant to therapy. OBJECTIVE: To correlate the results of serial phototesting in a patient with severe and refractory solar urticaria before and after treatment with plasma exchange. METHODS: Plasma exchange was performed five times over a period of 10 days. Phototesting to ultraviolet A (UVA) irradiation and visible light was performed with fluorescent ultraviolet tubes and an incandescent lamp. RESULTS: The urticaria that developed after very low light doses during baseline phototesting could not be provoked following plasma exchange. The patient is now almost symptom-free, with only occasional and transient hives more than 21 months after her last plasma exchange. CONCLUSIONS: Plasma exchange is a therapeutic modality to consider in highly light-sensitive patients when other treatments have failed. PMID- 10381947 TI - Point-counterpoint: electrolysis for permanent hair removal. PMID- 10381948 TI - Point-counterpoint: laser hair removal. PMID- 10381949 TI - A bullous eruption in an acutely ill child. PMID- 10381950 TI - Vibrio infections and rickettsioses: summary notes. PMID- 10381951 TI - Gene therapy and dermatology: more than just skin deep. AB - BACKGROUND: Recent advances in the molecular characterization of dermatologic disease have substantively augmented the understanding of the pathogenetic processes underlying disorders of the skin. This new knowledge coupled with progress in gene delivery technologies has paved the way for introducing cutaneous gene therapy into the dermatologic therapeutic armamentorium. OBJECTIVE: This review article includes an overview of the current strategies for delivery of gene therapy with an emphasis on the potential role of cutaneous gene delivery in the treatment of skin and systemic diseases. CONCLUSIONS: Accessibility for gene delivery, clinical evaluation, and topical modulation of gene expression render the skin a very attractive tissue for therapeutic gene delivery. However, there are several key hurdles to be overcome before cutaneous gene therapy becomes a viable clinical option. These include difficulties in inducing sustained expression of the desired gene in vivo, the challenge of targeting genes to long-lived stem cells, and the difficulty in achieving specific and uniform transfer to different compartments of the skin. However, these problems are not insurmountable and will likely be resolved in conjunction with ongoing advances in delineating gene expression profiles and other molecular properties of the skin, strategies for stem cell isolation, and improved approaches to regulating gene delivery and expression. These advances should create the framework for translating the enormous potential of cutaneous gene therapy into the clinical arena and, thereby, substantively improving the management of both cutaneous and systemic disease. PMID- 10381952 TI - Parry-Romberg syndrome with contralateral and ipsilateral extremity involvement. AB - BACKGROUND: The Parry-Romberg syndrome is an acquired progressive hemifacial atrophy involving the subcutaneous tissues of the scalp and face. Involvement of the extremities is uncommon in this syndrome. OBJECTIVE: A case of contralateral and ipsilateral extremity involvement in the Parry-Romberg syndrome is reported, and the difficulties in distinguishing this syndrome from linear scleroderma en coup de sabre are reviewed. METHODS: A MEDLINE search for cases of Parry-Romberg syndrome with contralateral extremity involvement was performed and the cases reviewed. RESULTS: Contralateral extremity involvement in the Parry-Romberg syndrome is rare. CONCLUSION: This may be the first case reported in the English literature of Parry-Romberg syndrome with contralateral and ipsilateral extremity involvement. PMID- 10381953 TI - Ultraviolet radiation-induced p53 responses in the epidermis are differentiation dependent. AB - BACKGROUND: The tumour suppressor, p53, is recognized as a crucial molecule in regulating cellular responses to various DNA-damaging agents. Very early on in the development of nonmelanoma cancers p53 is mutated or lost, suggesting that p53 is crucial in protecting normal keratinocytes from the harmful effects of ultraviolet (UV) radiation. OBJECTIVE: Using two mouse models, one with multiple copies of mutant p53 and the other a p53 "knockout," our laboratory has examined a role for p53 in UV-induced DNA damage and determined if these effects are differentiation dependent. CONCLUSION: We outline in this review a proposed model reflecting differentiation-dependent p53 regulation of UV-induced responses in keratinocytes. After exposure to UV, basal keratinocytes repair damaged DNA, whereas differentiating keratinocytes undergo cell death, both processes are regulated by p53. PMID- 10381954 TI - Emporiatric medicine--growing into the 21st century: from patient care to population care. PMID- 10381955 TI - Training, experience and interest of general practitioners in travel medicine in New Zealand. AB - BACKGROUND: In New Zealand, general practitioners (GPs) are a major group of travel health advisers. This study was designed to investigate the prevalence of training, experience, and interest in travel medicine or related areas, interest in undertaking training in travel medicine and how training might be best delivered. METHOD: Four hundred GPs were randomly selected from the register of the New Zealand Medical Council and sent self-administered questionnaires. Two reminders were sent. RESULTS: Three hundred and thirty-two (83%) GPs responded and these GPs advised an average of two travelers per week. Most GPs (257/282, 91%) reported that they had no training in travel medicine/related area. Training in travel medicine/related areas was significantly associated with age group (x2=14.09, df=6, p<.05), with the proportion of GPs with training in travel medicine/related area tending to be higher in the 45-49 and 50-54 years age groups, and also with GP college membership/fellowship (x2=6.39, df=1, p<.05). Forty-one percent (121/298) of respondents stated that they had previous experience working in tropical medicine/developing country. There was a significant association between GPs having experience working in tropical medicine/developing countries and training in travel medicine (x2=14. 19, df=1, p<.001) and those who were non-New Zealand graduates (x2=7. 84, df=1, p<.01). Forty-four percent (131/300) of respondents stated that they had an interest in travel medicine. Nearly two thirds of respondents (200/309, 65%) indicated that they would be interested in undertaking various types of travel medicine training, with a short course most commonly identified (159/309). The interest for training in travel medicine was significantly associated with those GPs with an interest in travel medicine (x2=26.45, df=1, p<.001), in younger age groups (x2=41.30, df=6, p<.001), a lower mean number of years since graduation (t value=5.70, df=297, p<.001), a higher mean proportion of patients who were travelers (t value=23.15, df=303, p<. 01), and a higher mean number of travelers seen per week (t value=22. 94, df=303, p<.01). The most common postgraduate qualification amongst GPs was membership/fellowship of a GP college (85/282, 30%), which was significantly more prevalent amongst the older age groups (x2=18.18, df=8, p<.05). Membership of travel medicine was very low. CONCLUSIONS: This cross-sectional study found that most GPs in New Zealand did not have any formal training in travel medicine, although more than two fifths of GPs indicated an interest in travel medicine and experience in tropical medicine/related area. GPs mainly wanted continuing medical education (CME) on travel medicine in the form of short and certificate level courses. As membership in GP colleges and other organizations was limited, other providers of CME should also be considered for providing more of these courses, such as universities and pharmaceutical companies. Providers of CME may target less experienced GPs and those GPs who may be seeing more travelers and use various approaches. Undergraduate and postgraduate medical curricula may also need to include more training in travel medicine. PMID- 10381956 TI - Knowledge of travel medicine providers: analysis from a continuing education course. AB - BACKGROUND: With millions of international travelers, there has been an increase in the scope and variability of travel medicine providers. A study was conducted to measure the baseline knowledge of providers, determine factors affecting this knowledge, and assess acquisition of knowledge after a continuing education course. METHODS: A one-day continuing medical education course was held for health care professionals interested in travel medicine. Prior to the course, attendees completed a test determining knowledge in malaria chemoprophylaxis, traveler's diarrhea management, vaccines, jet lag, the returned traveler, and other areas. An identical test was given after completion of the course. Performance on the test was analyzed by profession, area of specialty training, and experience in travel medicine. RESULTS: Seventy-seven attendees completed the precourse test. Forty-eight percent were physicians and 47% were nurses; 29% specialized in infectious diseases, 22% in occupational medicine and student health, and 18% in family or internal medicine; 60% had >/= 1 year of travel medicine experience while 20% had no experience. The precourse test score for all participants was 62.7% 6 6.5 (sd). Analysis by profession found that physicians scored the highest (71%). Providers with >/= 1 year of travel medicine experience scored higher than those with no experience (67% vs 53%, p <.01). Statistically significant correlations were found between precourse exam results and profession (+.432, p <.001) and travel medicine experience (+.365, p =.002). No significant correlation was found between precourse exam and area of specialty training. Combined mean score on the postcourse exam improved to 81.8% 6 4.5, an increase of 17.2% over the precourse score for those who took both tests (p <.001). CONCLUSIONS: The profession of the provider and the duration of experience in travel medicine were the most important correlations of baseline knowledge in travel medicine. All groups improved their knowledge following the course. PMID- 10381957 TI - Common health problems in HIV-infected travelers to the (sub)tropics. AB - BACKGROUND: A study was conducted to estimate the incidence of health problems in HIV-infected travelers with various degrees of immunodeficiency to the (sub)tropics. METHODS: A retrospective questionnaire-based study among HIV infected patients attending the outpatient department of a university hospital during three months in 1996 with a history of travel to (sub)tropical destinations in the proceeding 12 months. The outcome measures were incidences of and medical consultation rates for common travel-associated illnesses. RESULTS: Of 293 HIV-infected patients, 59 (20%) had traveled during the preceding 12 months; of these 36 (61%) responded. Ten (28%) had traveled more than once during this period. There were no significant differences between travelers and nontravelers regarding CD4 count and age. Fifteen respondents (42%) used cotrimoxazole (CTX) for PCP prophylaxis; 22 (61%) had sought pretravel health advice. Median duration of travel was 3 weeks. Respiratory infections were recorded by two respondents (6%, 95% confidence intervals 1-19%) and skin problems by 10 (28%, 14-45%). Of 31 respondents without diarrhea on departure, 10 recorded diarrhea (32%, 19-57%). The overall rate of medical consultation for travel-related complaints ws 5% abroad and 28% after return, respectively. There was no association between the risk of diarrhea and CD4 count or CTX prophylaxis, even after adjustment for differences in age, duration of travel and travel experience. CONCLUSIONS: Although numbers in this study are small, HIV-infected travelers tend to have a high rate of medical consultation for (possibly) travel related illness. Diarrhea is the most frequent complaint, but shows no strong association with degree of immunodeficiency. PMID- 10381958 TI - Travel health: perceptions and practices of travel consultants. AB - BACKGROUND: The global increase in international travel puts travelers at risk of travel-related morbidity and mortality. Prior to travel, most travelers have contact with a travel agency, thereby providing an opportunity for intervention. With this in mind we aimed to determine some of the travel-related health knowledge, practices and needs of travel consultants. METHODS: A cross sectional study was undertaken in which one travel consultant from each of 166 Western Australian travel agencies was asked to complete a self-administered questionnaire. RESULTS: One hundred and forty-five travel agencies (87%) agreed to participate in the study. Fifty-six percent indicated that they "usually" gave broad travel-related health guidelines and recommended their clients consult a medical practitioner. Almost all travel consultants reported discussing travel health insurance; very few provided information on sexually transmissible diseases, the risks associated with drug use, or first aid kits. Over 80% responded correctly to statements eliciting their knowledge on yellow fever, malaria, and food safety; the majority incorrectly answered questions on dengue fever and altitude sickness. Fifty-six percent of respondents thought that there was "not enough" readily accessible travel health information; 52% said they would like to be more involved in providing health information to their clients. CONCLUSIONS: Contact between travelers and travel agents offers an opportunity to promote awareness of travel-related health hazards. While travel consultants' health knowledge on some topics is adequate, in other areas it is inconsistent. Many travel consultants in Western Australia expressed a willingness to be involved in future health promotion activities. This participation may be best nurtured by providing travel consultants with: (1) better health education so they are able to identify high-risk travelers and destinations for medical referral; and, (2) health information in a format they feel comfortable distributing to their clients. PMID- 10381959 TI - Clinicoepidemiological study of imported malaria in travelers and immigrants to Madrid. AB - BACKGROUND: The number of Spanish travelers visiting malaria endemic areas, and the number of immigrants from malarial countries arriving in Spain are continuously increasing. However, little information about imported cases in Spain is available. METHODS: A prospective clinicoepidemiological study of imported cases of malaria diagnosed at a referral teaching hospital in Madrid, Spain. RESULTS: Of the 160 patients, sixty (37.5%) were immigrants and 100 (62.5%) Spanish nationals. Malaria was acquired in Africa by 98.3% of immigrants and in 83.0% of travelers. Falciparum malaria accounted for 71.8% of the cases, P. vivax for 11.9%, P. ovale for 10.6% and P. malarie for 5. 0%. Eleven (6.9%) patients, all immigrants, were asymptomatic. Severe complications were recorded in 17 (10.6%): 7, severe anemia; 3, cerebral malaria; 2, renal failure; 1, spontaneous splenic rupture; 1, acute pulmonary edema; 1, sepsis; 1, acute cerebrovascular accident; and 1, disseminated intravascular coagulation. There were no fatal cases. Among the 100 Spanish nationals, 44% did not follow any prophylaxis, 29% followed a correct prophylaxis, 27% were considered defaulters, and 39% took self-treatment without cure. CONCLUSIONS: There is a changing pattern of imported malaria in Madrid, with one third occurring in immigrants and two thirds in nationals. This data provides information about the reemergence of imported malaria to Europe. PMID- 10381960 TI - Travel advice given by pharmacists. AB - BACKGROUND: In 1993 more than 1 million Swiss residents traveled to a tropical or subtropical country. Although most pretravel advice is given by general practitioners, a number of travelers also seek advice from pharmacists. Little is known about the quantity and quality of travel advice given or the sources of information used by this group. METHODS: One-hundred and twenty randomly selected pharmacists from three Swiss cantons were first interviewed in a cross-sectional study on the telephone. All study participants subsequently received a pretested questionnaire, in which most of the questions asked on the phone were repeated, with additional questions regarding the sources of information used for travel advice and the cooperation of general practitioners. Included in both parts of the study were two scenarios of fictive travelers seeking health advice for destinations frequently visited by Swiss tourists (Thailand and Kenya). RESULTS: Of 136 pharmacists approached, all who said they sometimes gave travel advice, agreed to participate (88%). Fifty-six percent of them give travel advice regularly (mean 2-3 times per month). General knowledge on the main health hazards was good, but for treatment of travelers' diarrhea, only 59% spontaneously mentioned the need for increased fluid intake, whereas 100 % recommended antidiarrheal drugs. Protection from the sun was mentioned only by 10 % of the respondents, and only 8 % said that the traveler should seek advice from a medical doctor. Over 95% could name the three most important measures against mosquito bites, although up to 20% still recommend Vitamin B1 as well. On the telephone, only 19% (for Thailand) and 31% (for Kenya) gave accurate advice on malaria protection, and 13% (for Thailand) and 3% (for Kenya) could make correct recommendations about vaccination. However, more than 50% said that in practice they would consult documentation before giving any advice, with the Bulletin of the Federal Office of Health (BFOH) being the most commonly used source of information. In the questionnaire interview, where documentation was used, the accuracy of advice increased, especially for malaria protection (74% correct for Thailand and 93% for Kenya). CONCLUSIONS: The overall knowledge of Swiss pharmacists on travel medicine issues is satisfactory. Specific questions need further attention, such as treatment of travelers' diarrhea, sun protection and advice on malaria prophylaxis and vaccinations. For the latter two, clients should also consult a medical doctor. Collaboration between doctors and pharmacists, and the consistency of the advice given, are important in improving compliance. Reliable information sources are available in pharmacies and are used. PMID- 10381961 TI - GeoSentinel: the global emerging infections sentinel network of the International Society of Travel Medicine. AB - GeoSentinel is a network of 22 member travel/tropical medicine clinics (14 in the United States and 8 in other countries) initiated in 1995 by the International Society of Travel Medicine (ISTM). GeoSentinel is based on the concept that these clinics are ideally situated to effectively detect geographic and temporal trends in morbidity among travelers. The core surveillance tool is a single-page faxable form submitted to a central data site for each post-travel patient, including immigrants, refugees, and foreign visitors. Diagnoses are entered either as specific etiologies or as syndromes and are then linked to geographic locations, reference dates, and clinical presentations. In addition, electronic communication with the larger body of worldwide ISTM member clinics is periodically done to obtain broader data collection in response to specific inquiries. The scope of GeoSentinel has broadened from the initial vision of a provider-based sentinel network tracking emerging infections at their point of entry into developed countries. Its present goals are (1) to monitor global trends in disease occurrence among travelers; (2) to ascertain risk factors and morbidity in groups of travelers categorized by travel purpose and type of traveler; (3) to respond to urgent public health queries; (4) to develop educational priorities for travelers' health; and (5) to effect a rapid response by electronically disseminating alerts to surveillance sites, to all ISTM members in 55 countries, and to public health authorities. In addition, a major byproduct of the network, and now one of its strongest assets, has been the growth of partnerships between ISTM, Centers for Disease Control and Prevention and health care providers around the world, as well as other medical societies, government, and private organizations. The demographic data, travel patterns, and clinical presentations for the first 2813 patient records analyzed from the GeoSentinel sites are summarized in this paper. PMID- 10381962 TI - HIV and the returning expatriate. AB - Worldwide seroprevalence of the Human Immunodeficiency Virus (HIV) was estimated at 29.4 million at the end of 1996 with 75-85% of infections in adults transmitted through sexual intercourse, and by heterosexual intercourse in more then 70%. SubSaharan Africa is currently the area most heavily affected but over the past 5 years rapid spread of the virus has occurred in Asia. No developing country is free of HIV infection and HIV infection acquired abroad now accounts for most of heterosexually acquired HIV infection presenting in the (United Kingdom) UK, accounting for 25% of new HIV cases in the UK in the year to end March 1995 and 18% in the year to end March 1996, and at least 38% of new cases in Scotland in 1996. PMID- 10381963 TI - Vaccination of travelers against hepatitis A and B. AB - Despite the fact that effective preventive measures have become available, there has been no decline in the incidences of both hepatitis A and hepatitis B in most industrialized countries to date. This is, in part, due to the rapid increase in the number of travelers to areas of medium and high endemicity for both diseases, primarily developing countries. Targeting of travelers at risk of contracting these diseases for vaccination offers a chance of significantly reducing their incidence. Hepatitis A, an acute disease associated with poor food hygiene, is the most common vaccine-preventable infection in travelers. Hepatitis A immunity should, therefore, be considered essential for anyone visiting an area of high endemicity. In contrast, hepatitis B is a blood-borne virus which was thought, until recently, to pose a relatively low risk to the majority of travelers. However, the 1990s has seen international tourism and business travel grow faster in Europe than anywhere else in the world, with travel to areas of high endemicity for hepatitis B (Africa, Asia and South America) being commonplace. Thus the number of reported hepatitis B cases is increasing in many countries. Furthermore, there is considerable overlap of high-endemicity areas of hepatitis A and hepatitis B so that travelers are often considered to be at risk from both viruses. As well as separate hepatitis A and B vaccine preparations, a combined hepatitis A and B vaccine is now available which may offer improvements in vaccination schedule, enhanced patient compliance, and reduced cost. PMID- 10381964 TI - International tourists, motor vehicles and road safety: a review of the literature leading up to the Sydney 2000 Olympics. AB - The number of international visitors to Australia is forecast to rise to 4.6 million in the year 2000, the main attraction being the Olympic Games in Sydney. While the Olympic Games are expected to directly attract 132,000 international visitors as athletes, officials, judges, journalists and spectators, a further 1.49 million visitors will be attracted to Australia because of the global spotlight on the country in the years leading up to and during the staging of the event. Pre- and post-Games travel will also involve large numbers of tourists moving about in other Australian states and territories. For example, 50% of the extra tourists are expected to visit Queensland, 25% Victoria, 13% Western Australia and 9% the Northern Territory. In order to provide the best possible experience for all of these visitors, it is essential that potential causes of travel-related illness and/or injury are anticipated, and that procedures are established in each state to minimize harm and to deliver appropriate health care if it is required. Ideally, procedures and resources should be coordinated to achieve maximum effectiveness. PMID- 10381966 TI - Immune response to rabies booster vaccination in subjects who had postexposure treatment more than 5 years previously. PMID- 10381965 TI - Mefloquine for malaria chemoprophylaxis 1992-1998: a review. AB - Mefloquine is an orally administered blood schizontocide for the chemoprophylaxis of malaria in nonimmune travelers. New pharmacokinetic data has shown that food increases the bioavailability of mefloquine. Steady-state pharmacokinetics of weekly prophylaxis in long term travelers have shown that toxic accumulation does not occur and that weekly dosing is associated with protective levels of the drug. The pharmacokinetics of mefloquine are highly stereospecific and all pharmacokinetic parameters, except tmax are significantly different for the (+) and (-) enantiomers. Mefloquine and its metabolite are not appreciably removed by hemodialysis. Steady-state levels of mefloquine can be attained in a reduced time frame of 4 days compared to 7-9 weeks using a loading dose strategy of 250 mg mefloquine daily for 3 days followed thereafter by weekly mefloquine dosage. This strategy, is however, associated with a higher incidence of an adverse event (AE). Cumulative evidence suggests a high protective efficacy of mefloquine (>91%) in nonimmune travelers to areas of chloroquine resistant Plasmodium falciparum (CRPF) except for clearly defined regions of multi-drug resistance. Reports from sub-Saharan Africa indicate a low but increasing level of resistance to this drug. Mefloquine resistance is associated with halofantrine and quinine resistance but not with chloroquine resistance. Penfluridol has been shown to reverse P. falciparum mefloquine resistance in vitro. There is some controversy regarding the tolerabilty of mefloquine for malaria chemoprophylaxis. A review of the studies conducted during 1992-1998 shows that in the reporting of any AE the incidence lies in the range (12-90%) and where there is a comparator, is equivalent to the incidence reported for almost all alternative regimens. When some measure of subjective severity is applied to the rating of AE, it appears that 11-17% of travelers are, to some extent, incapacitated by AE. Major studies and worldwide monitoring have shown that serious events are rare. A recent meta analysis showed that rates of withdrawal and overall incidence of AE with mefloquine were not significantly higher than those observed with comparator regimens except that mefloquine was more likely to cause insomnia and fatigue. Withdrawals in mefloquine arms were higher than in placebo arms. No performance deficit or functional impairment was observed in five clinical toxicity studies of mefloquine prophylaxis, including a study of driving performance. There is limited data regarding use of mefloquine in pregnancy. Early animal studies have documented teratogenic and embryotoxic effects associated with the use of high dose mefloquine. Two studies have shown a relatively high incidence of spontaneous abortions in mefloquine users. Cumulative evidence, however, is reassuring and has led the World Health Organization (WHO) and the Centers for Disease Control and Prevention (CDC) to sanction the use of mefloquine in pregnant women during the second and third trimesters. In conclusion, mefloquine prophylaxis is recommended for travelers to high risk areas of chloroquine resistant Plasmodium falciparum. The risk of malarial infection and the proven efficacy of mefloquine to prevent malaria should be weighed against the risk of drug associated adverse events. PMID- 10381967 TI - Chikungunya in a North American traveler. PMID- 10381968 TI - Intradermal rabies vaccination and concurrent use of mefloquine. PMID- 10381969 TI - Delayed synchronous outbreak of Plasmodium vivax malaria in four travelers. PMID- 10381970 TI - Infectious diseases of Thailand. PMID- 10381971 TI - Malaria control in South Africa: symposium in the wilderness. PMID- 10381972 TI - Global warming and therapeutic agents during travel. PMID- 10381973 TI - GIDEON computer program for diagnosing and teaching geographic medicine. PMID- 10381974 TI - Sensitivity of three recent questionnaires to mild traumatic brain injury-related effects. AB - OBJECTIVE: To evaluate the sensitivity of three recently developed questionnaires to mild traumatic brain injury (MTBI)-related effects. DESIGN: Comparison of an MTBI group, within 3 weeks of injury, to a normal control group. SETTING: Outpatient rehabilitation clinic. SUBJECTS: 120 MTBI patients and 120 age, education, sex, and preinjury socioeconomic status-matched normal control subjects. MAIN OUTCOME MEASURES: The Problem Checklist (PCL) from the New York Head Injury Family Interview to assess MTBI symptoms, the Short Form-36 Health Survey (SF-36) to assess functional burden associated with health problems, and the Community Integration Questionnaire (CIQ) to assess home, social, and productive activities. RESULTS: Patients' self-ratings on the PCL and SF-36, but not the CIQ, were generally worse than those of the normal controls. The largest differences were obtained on SF-36 measures that appear to assess musculoskeletal injury effects. CONCLUSIONS: The PCL and SF-36 show promise as sensitive measures of MTBI-related effects. The SF-36 may be particularly useful in evaluating associated musculoskeletal injuries, which might otherwise be overlooked by MTBI health care providers. PMID- 10381975 TI - Neuropsychological impairments, vocational outcomes, and financial costs for individuals with traumatic brain injury receiving state vocational rehabilitation services. AB - OBJECTIVE: To determine the relationship among neuropsychological variables, vocational outcomes, and vocational costs for Missouri Division of Vocational Rehabilitation (MO-DVR) clients with traumatic brain injury (TBI). DESIGN: Clients referred for neuropsychological evaluations were followed until DVR case closure. Subjects were grouped according to the following DVR status at case closure: Successfully Employed, Services Interrupted, and No Services Provided. Spearman correlations with Bonferroni corrections were calculated to determine relationships among variables, and Kruskal-Wallis nonparametric one-way analyses of variance (ANOVAs) were conducted to evaluate differences in DVR group status in terms of neuropsychological variables and DVR costs. SETTING: All evaluations were completed through a Midwestern university neuropsychology laboratory. PATIENTS: 110 consecutively referred DVR clients with nonacute TBI referred for neuropsychological evaluation. MAIN OUTCOME MEASURES: Absolute level (ie, raw/standard scores) of neuropsychological functioning and relative degree of decline in: intelligence (WAIS-R), memory (WMS-R General and Delayed Memory Indices), attention (WMS-R Attention Index), speed of processing (Trails A), and cognitive flexibility (Trails B); DVR costs at closure. RESULTS: 1) Surprisingly, the Successfully Employed group had significantly greater neuropsychological impairments; 2) Greater decline in delayed memory was associated with higher DVR costs (r = -0.30, P <.05); and 3) More indices of relative decline were significantly correlated with vocational outcomes (5/6) than were indices of absolute functioning (3/6). CONCLUSIONS: DVR is effective in providing services to individuals with the most significant neuropsychological deficits; it is important to consider both absolute level of functioning and relative decline in functioning when evaluating TBI. PMID- 10381976 TI - A multimodal support group with Hispanic traumatic brain injury survivors. AB - OBJECTIVES: (a) To design and pilot a culturally sensitive and neuropsychologically informed support group addressing barriers to emotional, social, and vocational adjustment among high-level functioning Hispanic/Latino TBI survivors. (b) To determine efficacy through outcome measures. INTERVENTION: Ten-week multimodal, culturally sensitive support group focusing on TBI sequelae education, relaxation techniques, coping skills development, behavioral goal setting and monitoring, and family participation. PARTICIPANTS: Six Spanish speaking high-level functioning TBI survivors aged 20-42. SETTING: Outpatient neuropsychological assessment and treatment center. OUTCOME MEASURES: Beck Hopelessness Scale; Purpose in Life Test; Perceived Self-Regulatory Ability Inventory. RESULTS: Participants' sense of personal destiny and feelings of hopelessness improved, as evidenced by objective measures and self-report. A telephone interview a year later indicated that gains had been maintained, and most participants were vocationally active. CONCLUSIONS: Results underscore the importance of considering linguistic and ethnic factors in developing support groups. PMID- 10381977 TI - Utility of the functional assessment measure after discharge from inpatient rehabilitation. AB - OBJECTIVE: To assess the relationship between the Functional Independence Measure (FIM) and the Functional Assessment Measure (FAM), and community integration and return to work in patients with severe traumatic brain injuries (TBI). DESIGN: A cross-sectional, prospective design was used to collect data at 6 and 24 months postdischarge. The Return to Work Scale (RTW) and Community Integration Questionnaire (CIQ) were selected to assess return to work and community functioning. Predictor variables included the motor and cognitive subscales of the FIM and the FAM. SETTING: Follow-up database of an inpatient and community TBI Rehabilitation Unit. PARTICIPANTS: All consenting patients with TBI admitted to the unit, aged 16 or above. There were 88 patients at 6 and 79 patients at 24 month follow-up. RESULTS: At 6 months follow-up, the FAM and the FIM were roughly equivalent in their ability to predict RTW and CIQ scores. At 24 months, FAM motor was the only significant predictor of CIQ, and FAM cognitive scores displayed an advantage over the FIM in predicting employment status. CONCLUSIONS: The FAM subscales produced only modest gains in prediction of employment status and community integration at 24 months postdischarge. This may reflect ceiling effects on the functional measures, a limited range on the RTW measure, poor ecologic validity of functional disability measures in assessing handicap, or a combination of these factors. PMID- 10381978 TI - Distance education and caregiver support groups: comparison of traditional and telephone groups. AB - OBJECTIVE: To implement and evaluate the impact of telephone caregiver groups, compared with traditional face-to-face, on-site caregiver groups. DESIGN: Quasi experimental design comparing the two group types across time. SETTING: An urban tertiary rehabilitation hospital with a brain injury program servicing a vast geographical area. PARTICIPANTS: Caregivers of an adult person with a brain injury participated in either one of the 10 telephone groups (TGs) (N = 52 caregivers who completed a full set of research forms) or one of the 10 on-site groups (OGs) (N = 39 caregivers who completed a full set of research forms). The combined total from both groups was 91. INTERVENTION: Caregivers who lived within 40 km of the facility were placed in one of the OGs, and all others were placed in one of the TGs. Both types of groups met weekly for 9 to 10 weeks and were led by either social work or psychology professionals. The TGs met using teleconference technology. MAIN OUTCOME MEASURES: Profile of Moods States (POMS), Caregiver Burden Inventory (CBI), and the McMaster Model Family Assessment Devise (FAD), were administered 2 months before the first day of group, on the first day of group, on the last day of group, and 6 months after group. On the last day of group, a participant satisfaction survey was administered. RESULTS: There were similar amounts of improvements for the outcomes from OGs and TGs. Rural caregivers had fewer difficulties on all measures at all measurement intervals. In both types of group, participants showed a statistically significant improvement in POMS scores and a trend toward improvement in FAD and CBI results. Participants of both group types rated their experience highly, although rural caregivers were somewhat more satisfied. CONCLUSIONS: Telephone groups offer a method of providing support and education to rural caregivers that is as effective as traditional in-person OGs. PMID- 10381979 TI - Impact of clinically significant heterotopic ossification on functional outcome after traumatic brain injury. AB - OBJECTIVE: To assess heterotopic ossification's (HO) impact on functional outcome after TBI. DESIGN: Retrospective with matched control group, single center. SETTING: TBI Model System of Care at the Medical College of Virginia of Virginia Commonwealth University, Richmond, VA. PARTICIPANTS: Twenty-six patients with TBI and triple-phase bone scan confirmed HO were matched with 26 patients without clinical evidence of HO. MAIN OUTCOME MEASURES: Acute and rehabilitation lengths of stay (LOS), Admission and Discharge Functional Independence Measure (FIM) scores, FIM change, FIM efficiency (FIM gains per week), and discharge disposition. RESULTS: The two groups had similar acute care LOS. Patients with HO had significantly longer inpatient rehabilitation LOS and significantly lower FIM mobility and activities of daily living subscale scores on admission and discharge. FIM efficiency was significantly lower for the group with HO. Significantly fewer patients with HO were able to be discharged to home. CONCLUSIONS: HO is associated with a poorer functional outcome; however, it is not clear whether HO causes the decreased function or whether it may serve more generally as an indicator of those patients who will not progress as far or as rapidly during inpatient rehabilitation. PMID- 10381980 TI - Effect of cognitive rehabilitation on outcomes for persons with traumatic brain injury: A systematic review. AB - We evaluated evidence for the effectiveness of cognitive rehabilitation methods to improve outcomes for persons with traumatic brain injury (TBI). A search of MEDLINE, HealthSTAR, CINAHL, PsycINFO, and the Cochrane Library produced 600 potential references. Thirty-two studies met predetermined inclusion criteria and were abstracted; data from 24 were placed into evidence tables. Two randomized controlled trials and one observational study provided evidence that specific forms of cognitive rehabilitation reduce memory failures and anxiety, and improve self-concept and interpersonal relationships for persons with TBI. The durability and clinical relevance of these findings is not established. Future research utilizing control groups and multivariate analysis must incorporate subject variability and must include standard definitions of the intervention and relevant outcome measures that reflect health and function. PMID- 10381981 TI - Commentary: beyond statistics and research design. PMID- 10381982 TI - Commentary: cognitive rehabilitation outcomes. PMID- 10381984 TI - The authors respond PMID- 10381983 TI - Commentary: the validity of cognitive rehabilitation. PMID- 10381985 TI - Computerized assessment software update 1999. PMID- 10381986 TI - Rehabilitation: a luxury? PMID- 10381987 TI - From the editor PMID- 10381988 TI - Hyperkalemia and ionized hypocalcemia during cardiac arrest and resuscitation: possible culprits for postcountershock arrhythmias? AB - STUDY OBJECTIVE: Early countershock of ventricular fibrillation (VF) has been shown to improve immediate and long-term outcome of out-of-hospital cardiac arrest. However, studies indicate that countershock of prolonged VF most commonly results in asystole or a nonperfusing bradyarrhythmia (pulseless electrical activity [PEA]), which rarely respond to current therapy. The cause of these postcountershock rhythm disturbances is not well understood but may be related to electrical injury of the globally ischemic myocardium or to local metabolic abnormalities that impair impulse formation and cardiac contraction. The purpose of this study was to evaluate changes in serum potassium and free calcium homeostasis during cardiac arrest and advanced cardiac life support (ACLS) interventions. METHODS: After sedation, intubation, anesthesia, and instrumentation, VF was induced in 13 dogs. After 7.5 minutes of VF, animals were immediately countershocked, standard closed-chest CPR was initiated, and epinephrine was administered (1 mg in repeated doses if necessary). RESULTS: Ten animals could not be resuscitated despite 20 minutes of ACLS interventions. In these animals, a progressive increase in serum potassium was observed from the onset of ACLS to the termination of resuscitation efforts (4.3+/-.6 to 6.0+/-.8 mEq/L, P<.01). A significant increase was observed within 10 minutes of beginning ACLS measures. This was accompanied by a decrease in ionized calcium concentration over the same period (4.95+/-.40 to 3.44 mg/dL, P<.01). The decrease in ionized calcium was significant within 5 minutes of ACLS interventions. Nine of these 10 animals had either postcountershock asystole or PEA at the termination of resuscitative efforts. The increase in potassium was not related to acidemia. Successfully resuscitated animals did not demonstrate these electrolyte changes. CONCLUSION: Ionized hypocalcemia and hyperkalemia occur during prolonged resuscitative efforts and may be related to dysfunctional transcellular ionic transport mechanisms. These cations play important roles in cardiac electrical and contractile activity and may play a role in refractory postcountershock rhythm disturbances. PMID- 10381989 TI - The role of ipratropium bromide in the emergency management of acute asthma exacerbation: a metaanalysis of randomized clinical trials. AB - STUDY OBJECTIVE: This study was conducted to determine whether the addition of inhaled ipratropium to inhaled beta-agonist therapy is effective in the treatment of adults with acute asthma exacerbation. METHODS: Published reports of randomized, controlled trials assessing the use of ipratropium and beta-agonists in asthma were identified by a search of the MEDLINE, EMBASE, CINAHL, Biological Abstracts on CD, the Cochrane Library, and Current Contents databases. Bibliographies from identified studies and from review articles were manually searched. Published and unpublished reports in English, French, and Italian were identified and assessed for inclusion in the metaanalysis. Randomized, double blind, placebo-controlled trials were selected in which ipratropium was used as adjunctive therapy to beta-agonists in adult patients with acute asthma exacerbation presenting to a hospital emergency department or similar acute care setting. Data were extracted independently by 2 reviewers. For eligible trials, the mean percent change in peak expiratory flow rate (PEFR), or forced expiratory volume in one second (FEV1), and their SDs were assessed in the ipratropium treated and control groups. The effect of ipratropium on hospitalization rates and adverse effects were also analyzed. RESULTS: Data from 10 studies, reporting on a total of 1,377 patients with asthma, were pooled using a weighted average method. Compared with placebo, the use of ipratropium was associated with a pooled 7.3% improvement in FEV1 (95% confidence interval [CI] 3.8% to 10.9%), corresponding to an absolute improvement in FEV1 in the ipratropium/ beta-agonist group, which was 100 mL (95% CI 50 to 149 mL) above that seen for the group that received beta-agonist without ipratropium. Similarly, the pooled estimate of treatment effect in trials that reported data as PEFR was 22.1% (95% CI 11.0% to 33.2%), corresponding to an absolute peak expiratory flow improvement of 32 L/min (95% CI 16 to 47 L/min) in favor of the ipratropium/ beta-agonist combination group. When these data were combined using effect size as a common measure, the use of ipratropium was associated with a summary effect size of.38 (95% CI.27 to.48). Effect sizes were negatively correlated with baseline mean expiratory flows, suggesting that studies enrolling patients with more severe airflow obstruction showed greater absolute benefits of combination bronchodilator therapy. For the 3 trials reporting hospital admission data (n=1,031), patients receiving ipratropium had a relative risk of hospitalization of .73 (95% CI.53 to .99). The use of ipratropium was not associated with any severe adverse effects when used in conjunction with beta2 -agonists. CONCLUSION: There is a modest statistical improvement in airflow obstruction when ipratropium is used as an adjunctive treatment to beta2 -agonists for the treatment of acute asthma exacerbation. Although the clinical significance of this improvement in airflow obstruction remains unclear, it would seem reasonable to recommend the use of combination ipratropium/ beta-agonist therapy in acute adult asthmatic exacerbations, since the addition of ipratropium seemed to provide physiologic evidence of benefit without risk of adverse effects. PMID- 10381990 TI - Survey of health maintenance organization instructions to members concerning emergency department and 911 use. AB - STUDY OBJECTIVE: Anecdotal concerns suggest that health management organization (HMO) membership instructions may deter members from calling 911 or going to an emergency department for a perceived emergency. This study examines such instructions, specifically in regard to their definition of an emergency condition and associated instructions. METHODS: Member instructions were requested from 28 HMOs in 3 large West Coast cities with HMO penetration exceeding 30%. Fifteen (54%) provided membership materials. Features examined included the definition of an emergency, instructions for calling 911, specific instructions regarding chest pain and stroke, and mention of costs associated with emergency care. RESULTS: Instructions and definitions varied widely. Six HMOs (40%) included chest pain in their definition of an emergency; 2 (13%) included symptoms of stroke. Ten (67%) made mention of calling 911 or going to the ED somewhere within their instructions; 4 (27%) provided no options for calling 911 or seeking ED care. Three (20%) cited higher costs associated with ED care. Eleven (73%) indicated that claims would be denied for visits determined on retrospective review to be nonemergencies. CONCLUSION: Instructions varied considerably. Most did not include chest pain or symptoms of stroke in their definition of an emergency. Most did include directions to call 911 or go to an ED. Other instructions may lead members to call the HMO first during an emergency. PMID- 10381991 TI - Identification of randomized controlled trials from the emergency medicine literature: comparison of hand searching versus MEDLINE searching. AB - STUDY OBJECTIVE: As part of an ongoing project to identify all the randomized controlled trials (RCTs) in the emergency medicine literature, in association with the Cochrane Collaboration, 2 discrete studies were undertaken; the first, to compare motives for active participation in hand searching of the literature by emergency medicine professionals, and the second, to compare hand searching with MEDLINE searching of a number of emergency medicine journals. METHODS: All listed members of the British Association for Emergency Medicine (BAEM) and the Society for Academic Emergency Medicine (SAEM) received a standard letter outlining the objectives of the project, with 1 of 3 headings assigned on an alternate basis. Recruited volunteers hand searched journals prioritized from the emergency medicine literature. Each issue of each journal was hand searched for RCTs. In addition, a comprehensive MEDLINE search was conducted for each journal. The yields of RCTs from the 2 searching methods were compared for all journals and for each journal individually. RESULTS: No clear motivation for participation in this work could be ascertained because of the low response rates from BAEM and SAEM (10.1% and 1.8%, respectively). Only 18 (29.0%) of the 62 journals identified were indexed by MEDLINE. In the 14 journals indexed by MEDLINE for which hand searching was completed, a total of 710 RCTs were identified by a combination of the 2 approaches; of these, 592 (83.4%) were identified by hand searching alone and 483 (68.0%) by MEDLINE searching alone. Both methods identified 365 (51. 4%) RCTs; hand searching revealed an additional 227 (32.0%) that were not identified by MEDLINE searching, and MEDLINE searching found 118 (16.6%) that were not identified by hand searching. The difference between the proportions identified by hand searching and by MEDLINE searching (15.4%; 95% confidence interval [CI], 12.7% to 17.9%) was statistically significant (McNemar's chi2 test, 1 df, 33. 8; P <.0001). This difference was not significant for 8 of the journals. CONCLUSION: The response rates from mailing to members of the relevant professional organizations letters requesting participation in this work were very low and suggested that such an approach was not cost-effective. However, no formal costing exercise was undertaken. Searching results showed that, in the 14 emergency medicine journals indexed by MEDLINE for which hand searching was completed, hand searching led to identification of additional RCTs (primary articles) not found through MEDLINE searching. However, hand searching, although statistically significantly better than MEDLINE searching, failed to identify some of the RCTs found by MEDLINE searching, suggesting that hand searching is not a "gold standard" method and that the dual approach, promoted by the Cochrane Collaboration, may be the optimal approach for journals indexed by MEDLINE. PMID- 10381992 TI - Ingestion of low-concentration hydrofluoric acid: an insidious and potentially fatal poisoning. AB - STUDY OBJECTIVE: The purpose of this study was to provide the first description of the effects of ingestion of low-concentration hydrofluoric acid in a population reported to a regional poison control center. METHODS: A retrospective analysis of data collected by trained personnel using a standardized data collection system was performed. All charts involving hydrofluoric acid exposures for a 2-year period from a certified regional poison control center were identified by a computerized search. Each chart was abstracted by trained and blinded personnel. RESULTS: There were 1,772 exposures to hydrofluoric acid; 135 involved ingestion. There were 99 cases of human hydrofluoric acid ingestion for analysis. All ingestions involved consumer products containing 6% to 8% hydrofluoric acid. Symptoms, most commonly mild gastrointestinal effects, were reported by 49 patients. Two patients with minimal effects during an observation period of 2 to 4 hours deteriorated suddenly and died. All other patients recovered completely. Of 29 cases in which calcium concentrations were recorded, 4 cases of hypocalcemia occurred. All patients who had major effects or died were adults who had ingested more than 3 ounces of hydrofluoric acid with suicidal intent. Death occurred precipitously in patients who had appeared well a few minutes earlier. CONCLUSION: Death occurred in 2 patients, both of whom were adults who had ingested more than 3 ounces with suicidal intent. Ingestion of a household product containing hydrofluoric acid is a potentially life-threatening condition that requires close monitoring and prompt therapy. The abrupt deterioration and lack of warning signs indicate the need for better diagnostic methods. PMID- 10381993 TI - Double-blind, randomized study of nalmefene and naloxone in emergency department patients with suspected narcotic overdose. AB - STUDY OBJECTIVES: To compare the efficacy, safety, and withdrawal symptoms in emergency department patients with suspected narcotic overdose treated with nalmefene, an opioid antagonist with a 4- to 10-hour duration of action, with those treated with naloxone. METHODS: Adults in 9 centers who would otherwise receive naloxone for altered consciousness levels were randomly assigned to receive intravenous study drug (1 mg nalmefene, or 2 mg nalmefene or 2 mg naloxone, double-blinded) every 5 minutes as needed for up to 4 doses in a 4-hour study. Outcomes were 20-minute and 4-hour posttreatment changes in respiratory rates, Neurobehavioral Assessment Scale scores, Opioid Withdrawal Scale scores, and incidences of adverse events. RESULTS: Opioid positivity was recorded for 30 of 63 (1-mg nalmefene), 23 of 55 (2-mg nalmefene), and 24 of 58 (naloxone) cases, 75% of whom also had nonopioid central nervous system depressants. Most patients received only 1 dose of study drug. Similar, clinically meaningful improvements in respiratory rates and Neurobehavioral Assessment Scale scores were seen with all treatments. No statistical differences in efficacy or withdrawal outcomes were seen between treatment groups, and no significant overall time-treatment interactions occurred, in either the entire patient group or among opioid positive cases (P >.21, all comparisons). Adverse events occurred in 30.9% (2 mg nalmefene), 15.9% (1 mg nalmefene), and 15.5% (naloxone) of patients (P >.08); none were associated with morbidity. CONCLUSION: In this study of patients with varied potential causes of altered consciousness, nalmefene (1 mg and 2 mg) and naloxone (2 mg) appeared to be efficacious, safe, and to yield similar clinical outcomes. PMID- 10381994 TI - Agricultural avermectins: an uncommon but potentially fatal cause of pesticide poisoning. AB - STUDY OBJECTIVE: Avermectins have been used in the control of parasites and insects; however, human data concerning poisoning are lacking. This study investigated the clinical spectrum of avermectin poisoning. METHODS: A retrospective study was conducted to evaluate patients with avermectin poisoning reported to a poison center from September 1993 through December 1997. RESULTS: Eighteen patients with abamectin (Agri-Mek; 2% wt/wt abamectin) exposure and 1 with ivermectin (Ivomec; 1% wt/vol ivermectin) ingestion were identified. There were 14 male and 5 female patients, ranging in age from 15 to 83 years. Most patients were exposed as a result of attempted suicide (14). Oral ingestion (15) was the most common route of exposure. Four patients were asymptomatic, and 8 had minor symptoms after a mean ingestion of 23 mg/kg abamectin (4.2 to 67 mg/kg), or after dermal and inhalation contact. Seven patients manifested severe symptoms, such as coma (7), aspiration with respiratory failure (4), and hypotension (3), after a mean ingestion of 100.7 mg/kg avermectin (15.4 mg/kg for ivermectin and 114.9 mg/kg for abamectin). All 7 patients received intensive supportive care; 1 patient died 18 days later as a result of multiple organ failure. CONCLUSION: Ingestion of a large dose of avermectin may be associated with life-threatening coma, hypotension, and subsequent aspiration. PMID- 10381995 TI - Clinical decisionmaking based on venous versus capillary blood gas values in the well-perfused child. AB - STUDY OBJECTIVE: In children aged 1 month to 18 years, we sought to examine the correlation between venous and arterialized capillary blood gas values, and to determine whether the source of blood sample influenced the interpretation of the acid-base status and clinical management. METHODS: In a cross-sectional study, venous and capillary blood gas values were simultaneously obtained in acutely ill well-perfused patients treated in a pediatric emergency department. Intraclass correlation coefficients for capillary and venous measured gas values were calculated. Crude agreement and intraobserver concordance were calculated for responses of 2 intensivists to the interpretation and clinical management questions, based on capillary and venous gas results. RESULTS: Intraclass correlation coefficients for 78 capillary and venous paired measured gas values were.92 (pH), .80 (PCO 2 ), and .67 (PO 2 ). The alpha of concordance values between capillary and venous blood gas values, with 95% confidence intervals (CIs) were as follows, respectively, for physician A and B: interpretation, .61 (.47 to .73) and .48 (.41 to .55); need for bicarbonate,.85 (.73 to.97) and.80 (.72 to.88); and need for intubation .73 (.64 to .82), and .83 (.75 to .91). CONCLUSION: In the well-perfused patient, we believe that venous samples are an acceptable alternative to capillary blood samples for determination of blood gas values and for making clinical management decisions. PMID- 10381996 TI - Clinician assessment for acute chest syndrome in febrile patients with sickle cell disease: is it accurate enough? AB - STUDY OBJECTIVE: To determine whether the use of empiric chest radiography (CXR) is of significant value in detecting clinically unsuspected acute chest syndrome (ACS) in febrile patients with sickle cell disease (SCD). METHODS: Patients with SCD presenting to the emergency department and hematology clinic with temperature greater than or equal to 38 degrees C were prospectively evaluated using a physician-completed questionnaire. The questionnaire included inquiries into the patient's physical signs and symptoms and the physician's clinical impression for the presence of ACS. The questionnaire was completed before obtaining CXR results in all patients. RESULTS: Seventy-three patients with SCD with 96 febrile events were evaluated over a 1-year period. Twenty-four percent (23/96) of the patients had CXR evidence of ACS. On the basis of the questionnaire data, 61% (14/23) of ACS cases were not clinically suspected by the evaluating physician before obtaining CXR. Comparing the patients with and without ACS revealed that, with the exception of splinting (4/23 [17%] versus 0/73 [0%]), no symptom or physical examination finding helped to identify which patients had ACS. Fifty-seven percent of patients with ACS had completely normal findings on physical examination. The presentation of patients with clinically detected versus clinically unsuspected ACS also did not differ significantly. Length of hospitalization, oxygen use, and need for transfusion were the same in both the unsuspected and detected ACS groups. Overall physician sensitivity for predicting ACS was only 39%, and diagnostic accuracy did not improve significantly with increasing levels of pediatric training. CONCLUSION: ACS is common in patients with SCD who present with fever and was grossly underestimated by evaluating physicians. History and physical examination appear to be of little value in defining which febrile patients require CXR. In view of the mortality and morbidity associated with ACS, empiric CXR should be considered when evaluating a febrile patient with SCD. PMID- 10381997 TI - Do parents want to be present during invasive procedures performed on their children in the emergency department? A survey of 400 parents. AB - STUDY OBJECTIVES: No large study has addressed whether parents want to be present when invasive procedures are performed on their children in the emergency department. We conducted a survey to address this question. METHODS: The study used a self-administered, written survey consisting of 5 pediatric scenarios with increasing level of procedural invasiveness. Parents in an urban, teaching hospital ED waiting area were asked to participate. RESULTS: Of 407 persons asked to participate, 400 (98%) completed the survey. The number of parents expressing a desire to be present during a procedure performed on their child was 387 (97.5%) for venipuncture of the extremity, 375 (94.0%) for laceration repair, 341 (86.5%) for lumbar puncture, and 317 (80.9%) for endotracheal intubation. For a major resuscitation scenario, 316 (80.7%) wished to be present if their child were conscious during the resuscitation, 277 (71.4%) wanted to be present if their child were unconscious during the resuscitation, whereas 322 (83.4%) indicated a desire to be present if their child were likely to die during the resuscitation. Of the 400, 261 (65.3%) wished to be present for all 5 scenarios. Only 26 (6.5%) wanted the physician to determine parental presence in all 5 scenarios. CONCLUSION: Most parents surveyed would want to be present when invasive procedures are performed on their children. With increasing procedural invasiveness, parental desire to be present decreased. However, most parents would want to be in attendance if their child were likely to die, and nearly all parents want to participate in the decision about their presence. PMID- 10381998 TI - Nebulized ipratropium bromide in acute pediatric asthma: does it reduce hospital admissions among children presenting to the emergency department? PMID- 10381999 TI - The Cochrane Library: a resource for clinical problem solving in emergency medicine. PMID- 10382000 TI - Metaanalytic revision of endpoints. PMID- 10382001 TI - Managed care and emergency medical services: patient knowledge versus patient expectations. PMID- 10382002 TI - Pediatric case of accidental oral overdose of methotrexate. AB - Methotrexate is a chemotherapy antimetabolite, folic acid antagonist, that inhibits the enzyme dihydrofolate reductase resulting in decreased levels of tetrahydrofolate in the cells. This in turn blocks synthesis of thymidylate, a nucleotide necessary for DNA synthesis. It is readily absorbed from the gastrointestinal tract. Toxicity from overdose can affect multiple organ systems including bone marrow, liver, intestinal tract, kidneys, lungs, skin, and blood vessels, resulting in death in severe cases. Methotrexate is widely used to treat neoplastic disease, dermatologic disorders (psoriasis), and rheumatologic disorders (severe rheumatoid arthritis). As its indications for use increase, more accidental overdoses can be expected. We present the treatment and clinical course of one such case, that of a 2-year-old who accidentally took her grandmother's arthritis pills. Her initial serum level was 10 times greater than that needed to cause toxicity. She was treated with gastric lavage, activated charcoal, leucovorin rescue, and ICU admission. Her clinical course was unremarkable, and the only evidence of toxicity was a mild elevation in a liver associated enzyme that resolved without any clinical sequela. Leucovorin at a dose equal to or greater than the possible ingestion should be given as soon as possible in methotrexate overdoses. PMID- 10382003 TI - Neuromuscular blockade after ingestion of tree tobacco (Nicotiana glauca). AB - Two patients presented with life-threatening motor paresis after ingestion of leaves from the tree tobacco plant (Nicotiana glauca ). In addition to severe muscle weakness, bulbar palsies, flexor muscle spasm, hypertension, nausea, vomiting, and respiratory compromise were reported or observed. These are the fourth and fifth reported cases of a toxicologic emergency apparently caused by the alkaloid, anabasine, an isomer of nicotine found in the tobacco tree plant. The effects of this plant ingestion can mimic other better-known causes of paresis or paralysis. In areas of the country where the plant is indigenous, this toxicologic condition should be considered in the differential diagnosis of patients presenting with paresis or paralysis. PMID- 10382004 TI - Emergency medicine resident education: making a case for training residents to perform and interpret bedside sonographic examinations. PMID- 10382005 TI - Progress toward eliminating Haemophilus influenzae type b disease among infants and children--United States, 1987-1997. PMID- 10382006 TI - Commentary AB - [McCollough M: Progress toward eliminating Haemophilus influenzae type b disease among infants and children-United States, 1987-1997 [commentary]. Ann Emerg Med July 1999;34:110-111.] PMID- 10382007 TI - ER daydreams. PMID- 10382008 TI - Neurobiology of respiratory-pattern training in congestive heart failure. PMID- 10382009 TI - Orthopedic proceedings and dissertations, 1998. Abstracts. PMID- 10382010 TI - Increased funding bolsters FARAD; hotline to return. PMID- 10382011 TI - FDA: antimicrobial risk assessment model near completion. PMID- 10382012 TI - Vaccine-Associated Feline Sarcoma Task Force guidelines. Diagnosis and treatment of suspected sarcomas. PMID- 10382013 TI - Food safety group pushes for irradiation of precooked meats, more consumer education. PMID- 10382014 TI - USDA considers programs for farm-raised finfish. PMID- 10382015 TI - Disciplinary introversion in colleges of veterinary medicine. PMID- 10382016 TI - What is your diagnosis? Peritoneopericardial diaphragmatic hernia with herniation of mineralized fetuses. PMID- 10382017 TI - Revenue, expense, and returns on resources for US veterinary practices, 1995 and 1997. PMID- 10382018 TI - Veterinary human capital--knowledge, skills, and abilities that enhance employment opportunities outside private practice. PMID- 10382019 TI - Guidelines for development and application of aquatic animal health regulations and control programs. AVMA Aquaculture and Seafood Advisory Committee. AB - The creation of sound health regulations or disease control programs for any animal species is a complex endeavor. When the diverse stakeholder interests related to aquaculture are considered, this endeavor becomes daunting. The AVMA Aquaculture and Seafood Advisory Committee designed the following guidelines as a tool to assist aquatic animal health professionals who discuss potential regulations or control programs with government and industry entities. The guide focuses on determining whether a regulation or program is appropriate and, if so, developing a suitable and effective aquatic animal health plan. The Aquaculture and Seafood Advisory Committee was established in 1992 as an ad hoc committee of the AVMA Executive Board. The committee is composed of 9 veterinarians with diverse interests in aquaculture and seafood, and one non-veterinarian who represents the aquaculture industry. Participants from the USDA/APHIS and FDA serve as consultants to the Committee. PMID- 10382020 TI - Comparison of serum fructosamine and blood glycosylated hemoglobin concentrations for assessment of glycemic control in cats with diabetes mellitus. AB - OBJECTIVE: To correlate serum fructosamine concentrations with established measures of glycemic control and to compare serum fructosamine and blood glycosylated hemoglobin (GHb) concentrations as a means for assessing glycemic control in diabetic cats. DESIGN: Longitudinal cohort study. ANIMALS: 26 healthy cats, 5 cats with stress-induced hyperglycemia, 15 untreated diabetic cats, and 36 treated diabetic cats. PROCEDURE: Control of glycemia was classified and monitored and serum fructosamine and blood GHb concentrations were measured for 12 poorly controlled diabetic cats before and after improving glycemic control, 8 well-controlled treated diabetic cats before and after glycemic control deteriorated, and 5 cats with diabetes mellitus before and after onset of stress induced hyperglycemia. RESULTS: Mean serum fructosamine and blood GHb concentrations were significantly higher in untreated diabetic cats, compared with healthy cats, and in 24 poorly controlled diabetic cats, compared with 12 well-controlled diabetic cats. Mean serum fructosamine and blood GHb concentrations decreased significantly in 12 poorly controlled diabetic cats after improving glycemic control and increased significantly in 8 well-controlled diabetic cats after glycemic control deteriorated. A significant stress-induced increase in mean blood glucose concentration was evident 12 hours after insulin administration, but not in 5 docile diabetic cats that became fractious. CLINICAL IMPLICATIONS: Serum fructosamine and blood GHb concentrations are clinically useful tools for monitoring control of glycemia in cats with diabetes mellitus. PMID- 10382021 TI - Comparison of an amitraz-impregnated collar with topical administration of fipronil for prevention of experimental and natural infestations by the brown dog tick (Rhipicephalus sanguineus). AB - OBJECTIVE: To compare the efficacies of amitraz and fipronil for prevention of experimental and natural infestations of Rhipicephalus sanguineus. DESIGN: Prospective study. ANIMALS: 30 dogs. PROCEDURE: In 3 trials, dogs were allocated to 3 groups of 10 each. In trial 1, dogs were experimentally infested on day--1, and on day 0 were fitted with an amitraz-impregnated collar, treated topically with fipronil, or not treated. Ticks were counted daily until day 7, when viability of ticks and their progeny was determined. In trial 2, dogs were treated on day 0 and experimentally infested on days 7, 8, 10, and 13. Ticks were counted on days 8, 10, 13, and 18, and viability of ticks and their progeny was determined on day 18. In trial 3, dogs were exposed weekly to a tick-infested environment from day--3 to day 70. Dogs were treated on day 0, and ticks were counted and removed weekly from day 3 to day 77. RESULTS: Fipronil and amitraz were acaricidal and inhibited attachment and feeding. Amitraz had a significantly greater effect than fipronil on numbers of live, feeding ticks, egg hatchability, and larval viability, indicating partial ability to interrupt the tick life cycle. In field conditions, amitraz remained effective over the entire observation period. CLINICAL IMPLICATIONS: Amitraz had stronger and more sustained effects against tick infestation than fipronil. PMID- 10382022 TI - Serum total thyroxine, total triiodothyronine, free thyroxine, and thyrotropin concentrations in epileptic dogs treated with anticonvulsants. AB - OBJECTIVE: To determine whether administration of phenobarbital, potassium bromide, or both drugs concurrently was associated with abnormalities in baseline serum total thyroxine (T4), triiodothyronine (T3), free T4, or thyrotropin (thyroid-stimulating hormone; TSH) concentrations in epileptic dogs. DESIGN: Prospective case series. ANIMALS: 78 dogs with seizure disorders that did not have any evidence of a thyroid disorder (55 treated with phenobarbital alone, 15 treated with phenobarbital and bromide, and 8 treated with bromide alone) and 150 clinically normal dogs that were not receiving any medication. PROCEDURE: Serum total T4, total T3, free T4, and TSH concentrations, as well as serum concentrations of anticonvulsant drugs, were measured in the 78 dogs with seizure disorders. Reference ranges for hormone concentrations were established on the basis of results from the 150 clinically normal dogs. RESULTS: Total and free T4 concentrations were significantly lower in dogs receiving phenobarbital (alone or with bromide), compared with concentrations in clinically normal dogs. Administration of bromide alone was not associated with low total or free T4 concentration. Total T3 and TSH concentrations did not differ among groups of dogs. CLINICAL IMPLICATIONS: Results indicate that serum total and free T4 concentrations may be low (i.e., in the range typical for dogs with hypothyroidism) in dogs treated with phenobarbital. Serum total T3 and TSH concentrations were not changed significantly in association with phenobarbital administration. Bromide treatment was not associated with any significant change in these serum thyroid hormone concentrations. PMID- 10382023 TI - Chronic granulocytic leukemia in a dog. AB - A 4-year-old spayed dog had a recent history of increased WBC count and surgery for pyometra. Two weeks after surgery, WBC count was 57,640 cells/microliter; neutrophilia and immature myelocytic cells were detected. Histologic examination of liver and lymph node biopsy specimens revealed active granulopoiesis. Immature granulocytes that stained with chloroacetate esterase were evident. Bone marrow was excessively cellular and contained numerous granulocytes and blast cells. A diagnosis of chronic granulocytic leukemia was made on the basis of test results. Treatment with hydroxyurea returned the WBC count to reference range within 2 months. Mean survival time for dogs with chronic granulocytic leukemia is approximately 1 year; the dog of this report has remained healthy for more than 2 years. Chronic granulocytic leukemia is a rare neoplastic disease that must be differentiated from leukemoid inflammatory reactions. Although commonly described as a diagnosis determined by exclusion, diagnosis of chronic granulocytic leukemia should be made on the basis of specific criteria. PMID- 10382024 TI - Laparoscopically assisted resection of umbilical structures in foals. AB - A technique for laparoscopically assisted resection of umbilical structures in foals was developed. Eleven foals ranging from 8 to 42 days old underwent this procedure. Results of bacteriologic culture of umbilical structures were positive in 7 foals. Mean duration of anesthesia was 99 minutes, of which the initial 20 to 25 minutes were typically devoted to positioning and preparation of the foal for surgery. Major complications did not develop in any foal. Minor complications, such as slippage of the endoscopic ligating clip or laceration of the bladder during dissection, were dealt with successfully during the procedure. Potential benefits of use of laparoscopy (i.e., decreased postoperative morbidity, smaller incisions, and increased intraoperative access to structures) must be carefully weighed against the risks of increased duration of anesthesia. However, anesthetic-related complications were not observed in any foal. PMID- 10382025 TI - Use of mammary gland and colostral characteristics for prediction of colostral IgG1 concentration and intramammary infection in Holstein cows. AB - OBJECTIVE: To determine whether mammary gland or colostral characteristics at calving could be used to predict colostral immunoglobulin G1 (IgG1) concentration or intramammary infection (IMI) and whether leakage of colostrum affects IgG1 concentration. DESIGN: Prospective study. ANIMALS: 113 multiparous Holstein cows. PROCEDURE: Cows were examined within 3 hours of calving, and mammary gland and colostral characteristics, colostral volume, somatic cell count, and concentrations of IgG1, fat, and protein were determined. Bacteriologic culture of mammary secretions was performed approximately 14 and 7 days before calving and at calving. Associations of gland and colostral characteristics with colostral IgG1 concentration, colostral volume, and IMI were examined. RESULTS: Thick or thin colostrum had higher IgG1 concentration than colostrum of intermediate viscosity. Colostrum from mammary glands that were firm had low IgG1 concentration. Colostral IgG1 concentration was weakly correlated with volume. Intramammary infection was likely to be detected if colostrum contained clots or blood or if the California Mastitis Test (CMT) score was > or = 2. Somatic cell count was higher for glands with IMI than for uninfected glands, and CMT score was correlated with cell count. CLINICAL IMPLICATIONS: Mammary gland and colostral characteristics were of little value in predicting IgG1 concentration. Our findings do not support recommendations that first milking colostrum that is thin (watery) or that is from cows producing large volumes not be fed to dairy calves. Colostral characteristics, particularly CMT score, were of value for predicting IMI. PMID- 10382026 TI - Incidence of diarrhea among calves after strict closure and eradication of bovine viral diarrhea virus infection in a dairy herd. AB - OBJECTIVE: To determine whether strict closure of a dairy herd and eradication of bovine viral diarrhea virus (BVDV) infection would decrease incidence of diarrhea in calves during the first 31 days after birth, whether specific risk factors were associated with incidence of diarrhea in calves, and whether diarrhea was associated with weight gain of the calves. DESIGN: 2-year cohort study. ANIMALS: 448 calves. PROCEDURE: Calves were monitored from birth to 31 days of age. Fecal samples were tested for rotavirus, blood samples were tested for BVDV and antibodies to BVDV, and serum samples were tested for IgG concentration. Risk factors were evaluated by means of survival analysis. RESULTS: Incidence of diarrhea in calves decreased significantly after strict closure of the herd and eradication of BVDV infection. Risk factors for diarrhea in calves interacted in a multifactorial way. Rotavirus infection and low serum IgG concentration increased the risk that calves would develop diarrhea. Calves that developed diarrhea gained significantly less weight than calves that did not develop diarrhea. CLINICAL IMPLICATIONS: To control diarrhea among calves in a dairy herd, emphasis should be on maintaining a strictly closed herd free from BVDV infection. However, other measures, such as measures to prevent rotavirus infection and to ensure that calves receive an appropriate amount of colostrum after birth, should also be taken. PMID- 10382027 TI - Management of limb fractures in wapiti (Cervus elaphus): 22 cases (1993-1997). AB - OBJECTIVE: To determine treatment and outcome of a series of wapiti (elk) with fractures of the limbs. DESIGN: Retrospective study. ANIMALS: 22 wapiti. PROCEDURE: Medical records were reviewed to determine affected limb and bone, fracture configuration, method of treatment, outcome, and complications. RESULTS: 2 animals had fractures of the humerus; 8 had fractures of the radius, ulna, or both; 5 had fractures of the third metacarpal bone; 3 had fractures of the tibia; 2 had fractures of the femur; and 2 had fractures of the tarsal bones. Most fractures (n = 11) were closed, displaced, nonarticular fractures; 6 fractures were open. Four animals died or were euthanatized prior to fracture treatment, 2 were not treated because fractures had already healed, and 14 underwent fracture repair. In the remaining 2 animals, the affected limb was amputated. Five animals developed nonfatal complications (wound dehiscence, osteomyelitis [2 animals], delayed union, and malunion) and 2 developed fatal complications (gastrocnemius rupture and femoral fracture during recovery). Overall, 16 animals were discharged from the hospital, and all were doing well at follow-up, 2 months to 4 years after discharge. CLINICAL IMPLICATIONS: In wapiti, limb fractures can be successfully treated by means of internal or external fixation. The high rate of fracture healing, even among wapiti with open fractures, should encourage veterinarians to repair limb fractures in wapiti. PMID- 10382028 TI - Identification and management of an outbreak of Flavobacterium meningosepticum infection in a colony of South African clawed frogs (Xenopus laevis). AB - During the summer of 1996, an outbreak of Flavobacterium meningosepticum infection developed in a colony of South African clawed frogs (Xenopus laevis). Clinical signs were consistent with septicemia: ascites, anasarca, dyspnea, extreme lethargy, congestion of web vessels, petechial hemorrhages, and sudden death. Mortality rate reached 35%, and all infections were fatal. The organism was resistant to most antibiotics but was susceptible to enrofloxacin, chloramphenicol, and trimethoprim-sulfadiazine. Treatment with trimethoprim sulfadiazine was unsuccessful. Although the point source of the infection was not determined, several environmental reservoirs were identified, including a communal water barrel and various pieces of equipment. Molecular strain typing by pulsed-field gel electrophoresis and biochemical analyses revealed that frogs were infected with a single strain of F meningosepticum. Sanitation and management procedures were effective in controlling the outbreak. PMID- 10382029 TI - [Cyclosporine as an inhibitor of signal transduction. Molecular mechanisms of action]. PMID- 10382030 TI - [Isolation and characterization of Saccharopolyspora erythraea mutants, resistant to chloramphenicol]. AB - Formation of chloramphenicol resistant (CMr) spontaneous and nitroso-ethyl-urea induced mutants of S.erythraea, an organism producing erythromycin, was studied. The mutants differed by the level of the chloramphenicol resistance (10 to 40 micrograms/ml). Part of the chloramphenicol resistance mutations had a pleiotropic pattern. 63.8 per cent of the CMr mutants was characterized by the growth thermosensitivity and 18.9 per cent was characterized by the absence of the melanin production property. The antibiotic potency of 56 per cent of the CMr mutants was higher than that of the initial strain. In some of the CMr mutants the property of resistance to other antibiotics was changed. A higher resistance of some mutants to chloramphenicol correlated with their increased lincomycin resistance. It was shown that chloramphenicol induced resistance to kanamycin in S.erythraea 5. Such a property for the resistance induction was preserved in the spontaneous low CMr mutants while the high CMr mutants isolated after the culture exposure to the mutagen lacked it. The CMr S.erythraea phenotype was genetically instable. Two different amplifying DNA sequences (18 and 70-78 kb in size) were detected in the CMr mutant genome. The chloramphenicol resistance mutation was localized in the S.erythraea chromosome region restricted by markers ura1 and met1. PMID- 10382031 TI - [Feasibility of decreasing the adhesive activity of Corynebacterium diphtheriae by biopolymers of natural origin]. AB - Possible decreasing of the Corynebacterium diphtheriae adhesive activity by natural biopolymers was studied. It was shown that the strains of C.diphtheriae circulating on the Primorye Territory had middle, low or minimal adhesive activity. Natural biopolymers were found to decrease the adhesive properties of C.diphtheriae. The results of the study are promising for further investigation of natural biopolymers as agents preventing C.diphtheriae colonization on the stomatopharynx mucosa. PMID- 10382032 TI - [Investigation and identification of the antimicrobial component of the drug "Tomatol"]. AB - Antimicrobial activity of Tomatol against laboratory strains of Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 6538-P and Candida albicans ATCC 885-653 as well as against clinical isolates of Enterobacter spp., Streptococcus spp., Staphylococcus spp., Klebsiella spp. and Escherichia spp. was tested. Tomatol was shown to have a broad antimicrobial spectrum including gram-positive and gram negative organisms and Candida. The TLC investigation demonstrated that the Tomatol antimicrobial component was that of a complex of organic acids such as succinic, citric, tartaric and others. PMID- 10382033 TI - [Efficacy of meropenem in the treatment of severe complicated urinary tract infections]. AB - The clinical and bacteriological efficacies of meropenem in the treatment of 12 patients with urinary tract infection were studied. In 8 patients the drug was administered intravenously in a dose of 1 g every 8 hours and in 4 patients with the creatinine clearance below 50 ml/min it was administered in a dose of 1 g every 12 hours (the treatment course of 7 to 10 days). Meropenem was used in the monotherapy. Severe complicated urinary tract infections were mainly observed in the patients with long-term urolithiasis, subjected to repeated surgical interventions and isolating as a rule polyresistant strains of Pseudomonas aeruginosa and E.agglomerans as the pyelonephritis pathogens at a titre of 5 x 10(5)-5 x 10(8) microbial cells per 1 ml of the urine susceptible to meropenem in 80 to 96 per cent of the cases. The clinical efficacy of the drug was stated in all the patients while the bacteriological efficacy amounted to 88.9 per cent. PMID- 10382034 TI - [Treatment of intestinal dysbacteriosis in children with diabetes mellitus]. AB - The use of the probiotic Acylact in combination with Viferone containing human recombinant interferon-alpha 2 in the treatment of intestinal dysbacteriosis in children with insulin-dependent diabetes mellitus (IDDM) was shown to be highly efficient. Elimination of microecologic disorders in the intestine was accompanied in all the children by immune rearrangements. The quantity of secretory and serum immunoglobulins and complement increased and the white blood cell chemiluminescence proved to be higher. The correction of the immune status of the children with IDDM resulted in a certain decrease of glycemia. The data are indicative of the fact that such a treatment of intestinal dysbacteriosis in children with IDDM is promising. PMID- 10382035 TI - [Hospital infections caused by Pseudomonas aeruginosa. Spread and clinical importance of antibiotic resistance]. PMID- 10382036 TI - [Pharmacokinetic interactions between fluoroquinolones and methylxanthines]. PMID- 10382037 TI - [Microflora of the female genitalia and its sensitivity to antibacterial drugs]. PMID- 10382038 TI - [Selective suppression of polymerase chain reaction]. AB - The selective suppression of the polymerase chain reaction and methods based upon it (construction of cDNA libraries from low amounts of biological material, subtractive hybridization and differential display of mRNA, fast cloning of full size cDNA, chromosome walking, cloning in vitro, and others) are reviewed. These methods display a high effectiveness and, taken together, enable intricate DNA analyses to be performed--from the search for nontrivial sequences to the total sequencing of the corresponding genes. PMID- 10382039 TI - [Antibodies against synthetic peptide fragments of T-cadherin inhibit binding of low density proteins with T-cadherin]. AB - Potential antigenic determinants of the atypical lipoprotein-binding proteins T cadherin (p105) and its precursor (p130) from cells of human smooth muscles were synthesized by the solid phase method according to the Fmoc-scheme. These corresponded to the 51-61, 140-160, 161-179, 260-271, 340-352, 350-362, and 370 385 sequences of p130 and were chosen on the basis of computer analysis of its antigenic structure. The conjugates of the peptides with horseradish peroxidase were used for the immunization of mice and rabbits. Antisera against the peptides corresponding to the 140-160, 161-179, and 260-271 sequences of p105 were shown by immunoblotting to react with p105, which we isolated from the vascular cells of smooth muscles and earlier identified as T-cadherin. These antisera inhibited the binding of low density lipoproteins with p105 in a dose-dependent manner. These results confirmed the identification of the p105 protein as T-cadherin and demonstrated the fundamental possibility of studying the interaction of this protein with low density lipoproteins by using antipeptide antibodies that inhibit binding. PMID- 10382040 TI - [Prostaglandin-H-synthase: stability and activity during immobilization on silica gel]. AB - Prostaglandin H synthase was isolated in the form of microsomes from sheep vesicular glands and immobilized on silica gel. This system of prostaglandin synthesis was activated by calcium ions and stabilized by adrenaline. Microsomes immobilized in the presence of adrenaline and calcium ions were stable upon storage at 4 degrees C. After two months, their activity was 80% of the initial activity. Immobilized microsomes were able to catalyze several cycles of prostaglandin E2 synthesis with no substantial loss of activity: after eight utilization cycles, they retained 66% of their initial enzymic activity. PMID- 10382041 TI - [Cloning of streptavidin gene from Streptomyces avidinii and its expression in Escherichia coli. Secretion of streptavidin by E. coli cells]. AB - The streptavidin gene from Streptomyces avidinii was cloned, an expression plasmid constructed, and a highly effective strain producer of streptavidin created. It was shown that the leader peptide of streptavidin ensures the effective secretion of this protein into the periplasmic space of Escherichia coli cells. The degradation site of the leader peptide was detected. Upon treatment with the total fraction of proteases secreted by S. avidinii into the culture medium, "core" streptavidin was obtained, which retained the biotin binding function. PMID- 10382042 TI - [Synthesis and antiproliferative activity of a novel N-acyl derivative of doxorubicin]. AB - Doxorubicin was acylated with estrone 3-hemisuccinate. The modified derivative exhibited high antiproliferative activity in vitro toward cell cultures of the MCF-7 human mammary adenocarcinoma and HepG2 human hepatoma. PMID- 10382043 TI - [New breakpoints of t(9;22) translocation in chronic myeloid leukemia]. PMID- 10382044 TI - [Methodology of DNA observation under atomic force microscope]. PMID- 10382045 TI - Do the benefits of angiography outweigh the risks in the treatment of patients with carotid artery occlusive disease? PMID- 10382046 TI - The interventional radiographer. PMID- 10382047 TI - Interventional magnetic resonance: realistic prospect or wishful thinking? PMID- 10382048 TI - Transcatheter arterial infusion therapy in the treatment of advanced pancreatic cancer: a feasibility study. AB - PURPOSE: To evaluate the effects of transcatheter arterial infusion (TAI) therapy in 18 patients with advanced pancreatic cancer. METHODS: The drugs infused were epirubicin 60 mg, mitomycin C 20 mg, and 5-fluorouracil 500 mg. The efficacy of TAI was evaluated by a tumor marker (CA19-9), computed tomography (CT) findings, and postoperative histopathological specimens. RESULTS: In 10 of 15 cases, the tumor marker level was decreased after TAI therapy. In 6 of 14 cases, CT showed a decrease in the tumor size, and in 1 case, the tumor disappeared completely. In 6 cases the tumor could be resected. Necrosis, fibrosis, and degeneration of cancer cells were seen in 3 of 4 cases for whom a histopathological evaluation was done. The median survival was 11 months. In 17 patients back pain was the chief complaint, and was reduced to a self-controlled level in 10 patients following TAI therapy. No major complications were encountered. CONCLUSION: TAI appears to be an effective palliative treatment for advanced pancreatic cancer. PMID- 10382049 TI - Stages III and IV squamous cell carcinoma of the mouth: three-year experience with superselective intraarterial chemotherapy using cisplatin prior to definitive treatment. AB - PURPOSE: This study was designed to assess the 3-year experience with superselective intraarterial chemotherapy prior to definitive treatment for stages III and IV squamous cell carcinomas of the mouth. METHODS: Twenty-two patients prospectively received superselective intraarterial chemotherapy using relatively low-dose cisplatin via a transfemoral approach. The locations of the tumors were the tongue (n = 12), gingiva (n = 5), buccal mucosa (n = 2), hard palate (n = 1), floor of the mouth (n = 1), and lip (n = 1). After intraarterial chemotherapy, 21 patients underwent surgery (n = 14), radiation therapy (n = 6), or both (n = 1). The survival rate of 25 patients who underwent surgery with/without radiation therapy until 1992 at Kumamoto University Hospital was also evaluated as a historical control. The survival curve was calculated with the Kaplan-Meier method, and the statistical difference between survival curves was determined with the generalized Wilcoxon test. RESULTS: The overall response rate was 95% [complete response (tumor completely resolved), 24%; partial response (tumor reduction > or = 50%), 71%]. Fifty-two intraarterial infusions were performed without any catheter-related complications. Mild and transient local toxicity such as edema or mucositis of the infused area was relatively common. One patient died of renal failure from cisplatin. After a median follow up of 20 months (range 2-41 months), the estimated 3-year survival rate for patients who underwent intraarterial chemotherapy plus surgery was 91%. The survival of the patients who underwent intraarterial chemotherapy plus surgery tended to be longer than that of the historical control. CONCLUSIONS: Early tumor reduction without delay of subsequent treatments can be obtained by intraarterial chemotherapy while minimizing complications and possibly improving survival. Further investigations of long-term survival with larger series need to be performed. PMID- 10382050 TI - Solitary hepatocellular carcinoma fed by the cystic artery: limitation of transcatheter arterial embolization. AB - PURPOSE: To clarify the limitations of transcatheter treatment for hepatocellular carcinoma (HCC) with parasitic feeders from the cystic artery. METHODS: Three male patients had a solitary HCC (average diameter 3 cm) fed by the cystic artery among 221 patients with HCC from 1994 to 1997. One tumor was nourished entirely from the cystic artery arising from the medial branch of the left hepatic artery, and two tumors were fed partially by the cystic arteries arising from the anterior inferior branch of the right hepatic artery. We analyzed the indications for transcatheter treatment for these three patients. RESULTS: We chose not to embolize the cystic artery for fear of necrosis of the gallbladder. Although embolization of the anterior branch of the right hepatic artery was performed in one patient with a tumor fed partially by the cystic artery, only half the tumor was embolized. Two patients underwent hepatic resection, and one received percutaneous ethanol injection therapy. At follow-up of 28-40 months (average 33 months) all patients are alive. CONCLUSION: Feeding by the cystic artery represents a limitation of TAE for HCC. PMID- 10382051 TI - Venous sac embolization of pulmonary arteriovenous malformation: preliminary experience using interlocking detachable coils. AB - PURPOSE: To evaluate the indication and advantages of venous sac embolization of pulmonary arteriovenous malformations (PAVMs) using interlocking detachable coils (IDCs). METHODS: We performed percutaneous embolization in 12 PAVMs in four patients using IDCs, initially placed in the venous sac or at the feeding artery to prevent systemic migration of additional coils. We placed the IDCs in the venous sac in PAVMs with the following vascular architecture: the draining vein was larger than the feeding arteries and both vessels were interposed with the venous sac or there were short feeding arteries. RESULTS: Complete occlusion was achieved in all 12 PAVMs without significant complications. We deployed IDCs in the venous sac in eight PAVMs and in the feeding artery in four. CONCLUSION: Venous sac embolization may be beneficial in PAVMs with large out-flow vessels or short feeding arteries. IDCs are suitable for this procedure. PMID- 10382052 TI - Access-related venous stenoses and occlusions: treatment with percutaneous transluminal angioplasty and Dacron-covered stents. AB - PURPOSE: To determine the effectiveness of using Dacron-covered stents to treat access-related venous stenoses and occlusions. METHODS: Twenty-two Dacron-covered stents were placed in 20 patients: in the basilic or axillary vein (n = 2), cephalic vein (n = 3), subclavian vein (n = 5), and at the venous anastomosis of the polytetrafluoroethylene (PTFE) implant graft (n = 10). RESULTS: Initial technical success was 100%. The cumulative primary and secondary patency rates were 57% and 83% at 6 months, 29% and 64% at 12 months, and 29% and 53% at 18 months. A statistically significant difference in the stent patency was revealed by comparing the patients with stents in the subclavian vein and patients with upper arm stents. The secondary patency rates of the upper arm stents were 73% after 6, 12, and 18 months. CONCLUSIONS: Percutaneous placement of Dacron-covered stents is a safe and effective procedure for salvage of a dialysis fistula. First results are promising, with a tendency to prolongation of the time interval between reinterventions. PMID- 10382053 TI - Angiographic characteristics of symptomatic recurrent disease after infrainguinal percutaneous transluminal angioplasty. AB - PURPOSE: To evaluate the angiographic patterns of clinically manifest recurrent disease after infrainguinal percutaneous transluminal angioplasty (PTA) of stenoses and total occlusions. METHODS: Among 326 infrainguinal PTAs on 263 consecutive patients, selective angiography was performed on 61 limbs of 52 patients 1-60 months after the primary intervention because of clinically suspected recurrent disease. Lesion-specific and patient-related factors were analyzed for 75 angiographically confirmed recurrent lesions in 57 limbs of 48 patients. RESULTS: Recurrent disease was more frequently a stenosis when the original target lesion was a stenosis (92%, 44/48) than when the original lesion was a total occlusion (59%, 16/27; p < 0.001). When the original target lesion was a stenosis, the total length of the recurrent disease was longer than that of the original lesion [3.9 +/- 3.9 cm (mean +/- standard deviation) vs 2.8 +/- 2.7 cm; p = 0.03], while in the subgroup of original total occlusions the length of the recurrent lesion was shorter than that of the original occlusion (7.1 +/- 5.0 cm vs 9.9 +/- 6.9 cm; p = 0.02). Half the restenosis (22/44) extended beyond one or both ends of the original stenosis and 38% (6/16) of the reocclusions extended beyond the distal end of the original occlusion. CONCLUSIONS: The type of recurrent disease depends on the original lesion type and the restenotic lesion frequently extends beyond one or both ends of the original target lesion. PMID- 10382054 TI - Repositioning and leaving in situ the central venous catheter during percutaneous treatment of associated superior vena cava syndrome: a report of eight cases. AB - PURPOSE: To describe a combined procedure of repositioning and leaving in situ a central venous catheter followed by immediate percutaneous treatment of associated superior vena cava syndrome (SVCS). METHODS: Eight patients are presented who have central venous catheter-associated SVCS (n = 6 Hickman catheters, n = 2 Port-a-cath) caused by central vein stenosis (n = 4) or concomitant thrombosis (n = 4). With the use of a vascular snare introduced via the transcubital or transjugular approach, the tip of the central venous catheter could be engaged, and repositioned after deployment of a stent in the innominate or superior vena cava. RESULTS: In all patients it was technically feasible to reposition the central venous catheter and treat the SVCS at the same time. In one patient flipping of the Hickman catheter in its original position provoked dislocation of the released Palmaz stent, which could be positioned in the right common iliac vein. CONCLUSION: Repositioning of a central venous catheter just before and after stent deployment in SVCS is technically feasible and a better alternative than preprocedural removal of the vascular access. PMID- 10382055 TI - Fatal outcome in atrial migration of the Tempofilter. AB - PURPOSE: To report the risk of fatal atrial migration with the Tempofilter. METHODS: Among temporary filters, the high safety profile Tempofilter has been marketed as offering protection for up to 6 weeks. We implanted about 60 Tempofilters to prevent pulmonary embolism. The main indications were temporary thromboembolic risk, recurrent pulmonary embolism, and contraindication to or failure of anticoagulant therapy. Follow-up was performed regularly by plain abdominal film and Doppler ultrasound. Filters were removed about 4 weeks after placement. RESULTS: We encountered three cases (5%) of atrial migration and one case of 5-cm cephalad displacement of the filter. Of the three patients with atrial migration, two died within 3 days of implantation, one from a massive pulmonary embolism and the other with cardiac tamponade. One patient did not show any serious complications. CONCLUSIONS: The Tempofilter may actively migrate cranially and become dangerous in the case of migration within the heart. PMID- 10382056 TI - Effect of antithrombotic agents on the patency of PTFE-covered stents in the inferior vena cava: an experimental study. AB - PURPOSE: To evaluate the efficacy of antithrombotic agents in the prevention of stenosis of polytetrafluoroethylene (PTFE)-covered stents in the venous system. METHODS: Spiral Z stents covered with PTFE (PTFE-covered stents) were placed in the inferior vena cava (IVC) of 34 dogs. Nineteen dogs, used as a control group, were sacrificed at 2, 4, and 12 weeks. Fifteen dogs, previously given antithrombotic agents [cilostazol (n = 5), warfarin potassium (n = 5), cilostazol plus warfarin potassium (n = 5)] were sacrificed at 4 weeks, and then examined angiographically and histopathologically. The effect of the antithrombotic agents was compared between groups. RESULTS: The patency rate of the antithrombotic agent group was 93% (14/15), which was higher than the control group rate of 63% (12/19). The mean stenosis rate of the patent stent at both ends and at the midportion was lower at 4 weeks in the antithrombotic agent group than in the control group. In particular, the mean stenosis rate in the cilostazol plus warfarin potassium group was significantly lower than the control group (Tukey's test, p < 0.05). The mean neointimal thickness of the patent stent at both ends and at the midportion was thinner at 4 weeks in the antithrombotic agent group than in the control group. In particular, the thickness of the neointima in the cilostazol plus warfarin potassium group was significantly decreased when compared with the control group (Tukey's test p < 0.05). At 4 weeks, endothelialization in the antithrombotic agent group tended to be almost identical to that in the control group. CONCLUSION: The present study suggests that administration of an antithrombotic agent is an effective way of preventing the stenosis induced by a neointimal thickening of PTFE-covered stents in the venous system. PMID- 10382057 TI - Retrievable IVC square stent filter: experimental study. AB - PURPOSE: In vitro and in vivo evaluation of a new retrievable, home-made, inferior vena cava (IVC) Square stent filter (SSF) with two trapping levels. METHODS: In vitro, the SSF was compared in a flow model with the stainless steel Greenfield filter (SGF) for emboli-trapping efficiency by serially passing 300 emboli of 3 and 6 mm in diameter and 15-30 mm in length in each type of filter. Nine swine were used for the in vivo testing of the SSF for deployment and retrievability, emboli-trapping efficiency, stability, and self-centering ability and two were used (total of 11 swine) for testing repositioning and retrievability of the SSF at 2 weeks and for gross and histologic IVC changes at 2 months. RESULTS: In vitro, the SSF and SGF had similar efficiency in trapping large emboli but the SSF had significantly better efficiency than the SGF for trapping all sizes of emboli (91.7% vs 81%), medium size emboli (93% vs 80%), and small emboli (86% vs 69%). Efficiency decreased in both filters from the first to the fifth embolus in each series but was still significantly better for the SSF. With the SSF, 89% of emboli were caught at the primary and 11% at the secondary filtration level. In the nine animals used for acute studies, the SSF was easily placed in all 27 attempts, assumed a central position 26 times, and was easily retrieved in 21 of 22 attempts. One tilted filter needed additional manipulation for retrieval. During emboli injection in five swine, the SSF had 97.2% emboli trapping efficiency and demonstrated good stability. In the two animals used for longer-term evaluation, the filters were easily retrieved 2 weeks after implantation. Histologic evaluation at 2 months showed neointimal proliferation around the SSF wires in contact with the IVC wall, which was otherwise normal. CONCLUSION: The SSF is a promising filter. It is easy to place and retrieve, is stable after placement, and has high efficiency for trapping emboli. Promising results justify further experimental and eventual clinical studies with a commercially manufactured SSF. PMID- 10382058 TI - Post-traumatic pseudocyst of the spleen: sclerotherapy with ethanol. AB - We report a case of successful percutaneous treatment of a chronic post-traumatic splenic pseudocyst using alcohol as the sclerosing agent. A 26-year-old man presented with a symptomatic cystic mass located in the spleen. Aspiration of 300 ml of fluid was only temporarily effective, and therefore a drainage catheter was placed 3 days later. After histopathologic and microbiologic exclusion of a malignant or infectious origin, local sclerotherapy with alcohol was performed because of recurrence after percutaneous drainage. This therapy was repeated six times within 2 weeks. Two weeks later, the remaining volume was determined to be 16 ml. Six months after treatment the cyst was no longer visible. To our knowledge this is the first case of a chronic post-traumatic splenic cyst treated with alcohol. Percutaneous sclerotherapy of a symptomatic post-traumatic splenic pseudocyst may be an alternative to surgical treatment. PMID- 10382059 TI - Superior thyroid artery lesion after US-guided chemical parathyroidectomy: angiographic diagnosis and treatment by embolization. AB - A 71-year-old woman presented with a life-threatening thyroid hemorrhage after US guided chemical parathyroidectomy. The diagnosis was made by angiography followed by immediate embolization of a pseudoaneurysm of the left superior thyroid artery. Embolization controlled the hemorrhage, obviating the need for surgery. The patient made a full recovery with no evidence of further hemorrhage. Pseudoaneurysm of the superior thyroid artery is a rare cause of hemorrhage and percutaneous embolization is an effective method of treatment. PMID- 10382060 TI - Coronary-to-bronchial artery communication: report of two patients successfully treated by embolization. AB - We report two cases of coronary-to-bronchial artery communication responsible for coronary steal. In both cases the anastomosis originated from the proximal circumflex artery and developed because of bronchiectasis. In both cases closure of the anastomosis was achieved successfully by embolization. To date, the patients remained free from symptoms. PMID- 10382061 TI - Bronchial aneurysms mimicking aortic aneurysms: endovascular treatment in two patients. AB - Bronchial artery dilatation and aneurysm formation is a potential complication of local inflammation, especially in bronchiectasis. When the bronchial artery has an ectopic origin from the inferior segment of the aortic arch, aneurysms may mimick aortic aneurysms. Despite this particular location, endovascular treatment is possible. We report two such aneurysms that were successfully embolized with steel coils. PMID- 10382062 TI - Peroneal artery aneurysm treated by transcatheter coil embolization and temporary balloon occlusion in Behcet's disease. AB - Peroneal artery aneurysms in Behcet's disease have not been described to date. We present such a patient who was treated successfully using transcatheter embolization. PMID- 10382063 TI - A new method of thoracocentesis using CT guidance in patients with a small amount of pleural fluid. AB - A new technique of CT-guided diagnostic thoracocentesis (CT-TC) for patients with a small amount of pleural fluid was performed in 52 patients. More than 10 ml of pleural fluid was obtained successfully without any complications in all cases; 14 patients were found to have malignant cells in the pleural fluid. The main points of the CT-TC procedure are as follows: (1) The patient is placed supine with two radiolucent blocks underneath the shoulders and hips in order to make space for inserting the needle from the back (below). (2) Serial CT images are obtained to determine the insertion route and to measure the depth of the fluid level below the skin. (3) The needle is bent at the appropriate angle and length and is advanced upward slowly from the skin entry point on the back. CT-TC can also be used therapeutically in debilitated patients who can not maintain a sitting position or when the pleural fluid needs to be drained completely. PMID- 10382064 TI - Re: Creating a TIPS in a patient with difficult venous access: use of a nonfunctioning Denver shunt to access an occluded superior vena cava. PMID- 10382065 TI - Re: Treatment of early TIPS occlusion with a polytetrafluoroethylene (PTFE) covered zigzag stent. PMID- 10382066 TI - Re: Direct percutaneous coil embolization of a pseudoaneurysm under color Doppler guidance. PMID- 10382067 TI - Re: Combined arteriovenous malformation and aneurysm of the ulnar artery: successful arterial embolization by using absolute ethanol. PMID- 10382069 TI - Telomeres and telomerase: more than the end of the line. AB - Early studies of telomerase suggested that telomeres are maintained by an elegant but relatively simple and highly conserved mechanism of telomerase-medicated replication. As we learn more, it has become clear that the mechanism is elegant but not as simple as first thought. It is also evident that, although many species use similar, sometimes identical, DNA sequences for telomeres, these species express their own individuality in the way they regulate these sequences and, perhaps, in the additional tasks that they have imposed on their telomeric DNA. The striking similarities between telomeres in different species have revealed much about chromosome ends; the differences are proving to be equally informative. In addition to the differences between species that use telomerase, there are also a few exceptional organisms with atypical telomeres for which no telomerase activity has been detected. This review addresses recent studies, the insights they offer, and, perhaps more importantly, the questions they raise. PMID- 10382068 TI - Brain "embolism" detected by magnetic resonance imaging during percutaneous mitral balloon commissurotomy. AB - PURPOSE: The common finding of thrombi between the bifoil balloons when they were extracted after mitral dilation prompted us to look for evidence of minor brain embolisms using the sensitive technique of BMRI (brain magnetic resonance T2 weighted imaging). METHODS: BMRI was performed within 48 hr before and after a percutaneous mitral balloon commissurotomy (PMBC) in each of the 63 patients in this study. RESULTS: There was evidence (hyperintensity foci: HI) of a previous asymptomatic brain embolism in 38 of 63 patients before PMBC and a new HI appeared in 18 of 63 patients after the procedure. New HI signals were found exclusively in the white matter in 8 of 18 patients and in only 3 of 18 were HI signs larger than 1 cm. One patient, with an HI signal > 1 cm in the thalamus and another < 1 cm in the brain stem, presented diplopia accompanied by other minor clinical signs. The differences in HI rate among four subgroups (1, older vs younger than 43 years; 2, sinus rhythm vs atrial fibrillation; 3, echo score < 8 vs > 8; 4, patients from western countries vs the others) were not statistically significant, probably because the number of patients in each subgroup was low. Patients in atrial fibrillation had slightly more (not significant) HI before PMBC (15/20, 75%) than patients in sinus rhythm (23/43, 53%), but after PMBC their HI frequencies were similar (atrial fibrillation: 5/20, 25%; sinus rhythm: 13/43, 30%). CONCLUSION: Brain microembolism is frequent during PMBC, but is often anatomically limited and free from clinical signs in most cases. Brain embolism seems to be related mainly to the procedure itself and not the features of the patient. PMID- 10382070 TI - The nuclear distribution of Polycomb during Drosophila melanogaster development shown with a GFP fusion protein. AB - The chromatin protein Polycomb (PC) is necessary for keeping homeotic genes repressed in a permanent and heritable manner. PC is part of a large multimeric complex (PcG proteins) involved in generating silenced chromatin domains at target genes, thus preventing their inappropriate expression. In order to assess the intranuclear distribution of PC during mitosis in different developmental stages as well as in the germ line we generated transgenic fly lines expressing a PC-GFP (Green Fluorescent Protein) fusion protein. Rapidly dividing nuclei were found to display a rather homogeneous PC-GFP distribution. However, with increasing differentiation a pronounced subnuclear pattern was observed. In all investigated diploid somatic tissues the bulk of PC-GFP fusion protein is depleted from the chromosomes during mitosis: however, a detectable fraction remains associated. In the male germ line in early spermatogenesis, PC-GFP was closely associated with the chromosomal bivalents and gradually lost at later stages. Interestingly, we found that PC is associated with the nucleolus in spermatocytes, unlike somatic nuclei. In contrast to mature sperm showing no PC GFP signal the female germ line retains PC in the germinal vesicle. PMID- 10382071 TI - The association of ATR protein with mouse meiotic chromosome cores. AB - The ATR (ataxia telangiectasia- and RAD3-related) protein is present on meiotic prophase chromosome cores and paired cores (synaptonemal complexes, SCs). Its striking characteristic is that the protein forms dense aggregates on the cores and SCs of the last chromosomes to pair at the zygotene-pachytene transition. It would appear that the ATR protein either signals delays in pairing or it is directly involved in the completion of the pairing phase. Atm-deficient spermatocytes, which are defective in the chromosome pairing phase, accumulate large amounts of ATR. The behaviour of ATR at meiotic prophase sets it apart from the distribution of the RAD51/DMC1 recombinase complex and our electron microscope observations confirm that they do not co-localize. We failed to detect ATM in association with cores/SCs and we have reported elsewhere that RAD1 protein does not co-localize with DMC1 foci. The expectation that putative DNA damage checkpoint proteins. ATR, ATM and RAD1, are associated with RAD51/DMC1 recombination sites where DNA breaks are expected to be present, is therefore not supported by our observations. PMID- 10382072 TI - Functional compartmentalization of the nucleus in the budding yeast Saccharomyces cerevisiae. AB - By combining cryofixation and cryosubstitution in a structural and functional analysis of the nucleus of Saccharomyces cerevisiae, we identified morphological subcompartments in the nucleolus. These were similar to those of nucleoli of higher eukaryotes, such as the fibrillar centre (FC), the dense fibrillar component (DFC) and the granular component (GC). In situ hybridization and immunocytochemistry revealed RNA polymerase I and proteins involved in early steps of ribosomal maturation along the DFC, while the ribosomal genes were detected at the FCs. Our results also suggest that ribosomal transcripts are distributed along a nucleolar network that might include both DFC and GC. We also show that pre-ribosomal subunits may be exported along tracks to the cytoplasm. Export takes place through all the pores of the nuclear envelope, not just those in contact with the nucleolus. Moreover, comparison of the nucleolar organization in S. cerevisiae and in Schizosaccharomyces pombe demonstrated than the distribution of the 5S genes with respect to the 35S transcription unit does not modify the organization of the nucleolus. We also report, for the first time, the ultrastructural localization of RNA polymerase II in yeast. The distribution of RNA polymerase II and morphological details that could be observed in the extra nucleolar region of cryofixed cells provided cytological evidence of a peripheral region extending along the nuclear envelope that could correspond to heterochromatin in higher eukaryotes. PMID- 10382073 TI - Broken chromosomal ends can be elongated by conversion in Drosophila melanogaster. AB - The fate of the termini of X chromosomes broken in the regulatory region of the yellow gene was followed in heterozygotes with X chromosomes carrying a point mutation inactivating the yellow gene. Each generation had a loss of about 70 terminal base pairs from the broken chromosome. However, gene conversion restoring the correct sequence at the chromosomal terminus took place with a frequency of about 1 x 10(-2) per generation. The average length of the conversion track was 2.7 kb. No recombination events occurred. In addition, we found that the normal functioning of the yellow body and wing enhancers located at the tip of the chromosome required about 4 kb of additional upstream sequence. PMID- 10382074 TI - The degradation profile of extrachromosomal circular DNA during cisplatin-induced apoptosis is consistent with preferential cleavage at matrix attachment regions. AB - Extrachromosomal circular DNA molecules are prevalent in cancer cells and harbor amplified genes, such as oncogenes and drug resistance genes, that can provide a selective growth advantage to cancer cells. These circular DNA structures include double minute chromosomes (dmin), which can be detected with light microscopy following Giemsa staining, and submicroscopic circular DNA structures referred to as episomes. In this study, we investigated the fate of dmin and episomes in multidrug-resistant human epidermoid KB-VI cells undergoing cisplatin-induced apoptosis-a mode of cell death initially characterized by the fragmentation of chromosomal DNA, while the nuclear membrane remains intact. The circular DNA structures carry amplified copies of the multidrug resistance gene (MDR1). During cisplatin-induced apoptotic cell death, episomes and dmin, as well as native chromosomes, were degraded into high molecular weight DNA fragments of approximately 50 kb in length. DNA fragments in this size range appear to result from the preferential cleavage of matrix-associated regions in chromatin with the subsequent release of 20-30 nm loop domains of chromatin from the nuclear scaffold. Scanning electron microscopy studies were performed and confirmed the presence of 30 nm filaments in a higher-order DNA packing of MDR1-containing dmin and episomes. These combined data provide strong evidence that the higher-order DNA packing of episomes, as well as dmin, is similar to that of native chromosomes and underscore the potential for extrachromosomal DNA amplicons to study the structural and functional organization of chromatin. We discuss the implications of extra-chromosomal DNA matrix associated regions competing with native chromosomal DNA for binding to the nuclear matrix in tumor cells. PMID- 10382075 TI - Identification of coiled body-like structures in meristematic cells of Pisum sativum cotyledonary buds. AB - This study focused on two types of nuclear bodies visible in plant cells that were previously thought to be similar to the coiled bodies (CBs) of animal cells: the nucleolus-associated body (NAB) and dense body (DB). We show that both NABs and DBs share common features with animal CBs: they consist of ribonucleoproteins, are silver-stainable, and lack DNA. Immunoelectron microscopy shows that only the NABs are rich in snRNAs and fibrillarin, two markers characteristic of animal CBs. This suggests that NABs rather than DBs are the plant counterparts of the CBs of animal cells. These structures appear most frequently in cells blocked in G0-1, their frequency gradually declining with resumption of the cell cycle and nucleolar activity. During this reactivation period, NABs are released from the nucleolus to the nucleoplasm, suggesting that they may act as nuclear transport or sorting structures. PMID- 10382076 TI - Updated estimates of the effectiveness of the Yuzpe regimen of emergency contraception. AB - The purpose of this study was to provide revised estimates of the effectiveness of the Yuzpe method of emergency contraception. Through a literature search, we identified eight studies that present the number of women treated and outcome of treatment by cycle day of unprotected intercourse relative to expected day of ovulation. Using five sets of external estimates of conception probabilities by cycle day of intercourse among women not using contraception, we assessed the effectiveness of the Yuzpe regimen. The 45 estimates of effectiveness, based on eight separate studies and the eight studies combined and five different sets of conception probabilities by cycle day, ranged from a low of 56.4% to a high of 89.3%. Our preferred point estimate is that the Yuzpe regimen reduces the risk of pregnancy by 74.1%, with a 95% confidence interval extending from 62.9% to 79.2%. True effectiveness is likely to be > 74% because treatment failures (observed pregnancies) include women who were already pregnant when treated and women who became pregnant after being treated. PMID- 10382077 TI - Outcomes of suction curettage and mifepristone abortion in the United States. A prospective comparison study. AB - A prospective, nonconcurrent cohort analysis of 178 mifepristone/misoprostol and 199 suction curettage abortion subjects, ages > or = 18 years, with intrauterine pregnancies < or = 63 days estimated gestational age, was conducted to compare the outcomes of suction curettage abortion to those of medical abortion. The medical abortion subjects received 600 mg of mifeprisone orally, followed by 400 micrograms of oral misoprostol 2 days later. Surgical abortion subjects underwent electronic vacuum aspiration. All subjects were followed for 2 weeks or until the absence of clinical bleeding. Outcome measures included a successful abortion (complete abortion without a surgical intervention), duration of bleeding, and morbidity. Overall, 18.3% medical and 4.7% surgical patients failed their primary procedure and received an unanticipated suction curettage (RR 3.93, 95% CI 1.87, 8.29). Four mifepristone subjects required curettage for acute bleeding, nine to manage ongoing pregnancy, and five for incomplete abortion. Fourteen mifepristone and eight surgical subjects required curettage for persistent bleeding. The median time delay for therapeutic curettage was significantly longer in the medical abortion group (35 versus 8 days; Mann-Whitney U = 17.0, p = 0.008). Medical subjects experienced significantly longer bleeding (mean difference = 9.6 days, 95% CI 6.8, 12.4). No significant change in hemoglobin occurred in either group. Mifepristone patients reported significantly greater pain (77.1% vs 10.5%; RR 7.4, 95% CI 4.7, 11.5), and nausea or vomiting (68.6% vs 0.6%; RR 117.9, 95% CI 16.7, 834.7). Women receiving mifepristone/misoprostol are more likely to require an unplanned surgical intervention than women who undergo suction curettage. They experience more discomfort with their procedure and in the follow up interval, bleed for a longer period, and remain at risk for surgical completion curettage for several weeks. PMID- 10382078 TI - Clinical comparison of triphasic norgestimate/35 micrograms ethinyl estradiol and monophasic norethindrone acetate/20 micrograms ethinyl estradiol. Cycle control, lipid effects, and user satisfaction. AB - This six-cycle, multicenter, open-label, randomized study compared the clinical experience of two low-dose oral contraceptives (OC): a triphasic OC containing norgestimate (NGM) and 35 micrograms ethinyl estradiol (EE) (Ortho Tri-Cyclen) and a monophasic OC containing norethindrone acetate (NETA) and 20 micrograms EE (Loestrin Fe 1/20). Cycle control, lipid and androgen profiles, and user satisfaction were studied in new-start OC users (i.e., no prior use within 60 days). Breakthrough bleeding or breakthrough spotting (BTB/BTS) occurred in a significantly smaller percentage of NGM/EE users than NETA/EE users during each of six cycles (p < or = 0.002). The incidence of BTB/BTS ranged from 3.7% to 13.5% for NGM/EE users and from 23.5% to 49.7% for NETA/EE users. Significantly fewer NGM/EE users than NETA/EE users experienced absence of menses at cycles 2 through 6 (p < or = 0.003). The percentages of women having no menses at each cycle ranged from 0.9% to 4.7% for NGM/EE users and from 10.3% to 21.3% for NETA/EE users. NGM/EE users reported a significantly (p < 0.001) higher level of satisfaction with their OC at the end of six cycles than did NETA/EE users, but there was no significant difference in compliance, discontinuation rates, or adverse events between the two groups. NGM/EE produced a significantly (p < or = 0.001) greater beneficial effect on HDL-C, HDL2, and apo A-I than did NETA/EE. No statistically significant treatment differences were found for total cholesterol, LDL-C, triglycerides, or apo-B. Both OC increased sex hormone binding globulin and decreased free testosterone, but NGM/EE had a significantly greater effect (p < 0.009). PMID- 10382079 TI - A pilot efficacy study with a single-rod contraceptive implant (Implanon) in 200 Indonesian women treated for < or = 4 years. AB - In this 4-year open-label, noncomparative, single-center pilot efficacy study, the contraceptive efficacy, safety, bleeding pattern and acceptability of Implanon was studied in 200 sexually active women of proven fertility in Indonesia. All subjects received the single-rod subdermal implant Implanon, which contains 68 mg etonogestrel (3-keto-desogestrel), with an initial release rate of 67 micrograms etonogestrel/day. Contraceptive efficacy was analyzed by calculation of the pregnancy rate, bleeding patterns were determined by the 90 day reference period method, and acceptability by the discontinuation rate. No in treatment pregnancies were reported during 658.4 women-years of exposure, resulting in a Pearl Index of 0.0 (95% CI 0.0-0.6). The overall bleeding pattern was acceptable, with no discontinuations because of irregular bleeding. Incidence of irregular bleeding was highest during the first two reference periods and decreased thereafter. Amenorrhea was experienced by 7%-12% of subjects during years 1 and 2, by 5%-7% during year 3, and by 2%-5% during year 4, with one discontinuation because of amenorrhea. No clinically significant changes were reported for systolic and diastolic blood pressure, body mass index, and hemoglobin level. Three adverse experiences were related to treatment and resulted in discontinuation (two headaches and one dyspnea). One difficult implant removal was reported. In conclusion, this pilot efficacy study indicates that Implanon provides excellent contraceptive reliability and an acceptable bleeding pattern. Overall safety and acceptability are good, as suggested by the low incidence of adverse experiences and the low discontinuation rate. PMID- 10382080 TI - Interleukin-6 and tumor necrosis factor-alpha concentrations in the intrauterine cavity of postmenopausal women using an intrauterine delivery system releasing progesterone. A possible mechanism of action of the intrauterine device. AB - Intrauterine devices (IUD) provide effective contraception. The current study evaluates the concentration of interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha) in the intrauterine fluid of postmenopausal women using an intrauterine delivery system releasing progesterone (IDS-P). Intrauterine fluid was obtained by lavage, and IL-6 and TNF-alpha were analyzed using an enzyme linked immunosorbent assay (ELISA). Statistical analysis was performed with a one way analysis of variance (ANOVA). Intrauterine fluid IL-6 levels were 33.6 vs 6.09 pg/sample IDS-P vs no IDS-P (p = 0.0301). Intrauterine TNF-alpha levels for women using the IDS-P were higher than in nonusers, but the differences did not reach statistical significance. IL-6 and TNF-alpha levels were increased in the intrauterine cavity of postmenopausal women with an IDS-P. These data suggest that secreted cytokines could be a potential mechanism of IUD contraceptive efficacy. PMID- 10382081 TI - Vasectomy and subsequent cardiovascular disease in US physicians. AB - Due to previous animal research suggesting accelerated atherosclerosis following vasectomy, we examined whether vasectomy increases the risk of subsequent cardiovascular disease (CVD), including myocardial infarction (MI), angina pectoris, coronary revascularization, and stroke, in the US Physicians' Health Study. Of 22,071 US male physicians participating in the study, aged 40 to 84 years at entry and free from cardiovascular disease and cancer, 21,028 reported on the 60-month questionnaire whether they had undergone vasectomy prior to randomization. Of the 4546 physicians with vasectomy, 1159 had undergone the procedure at least 15 years before entry. During 258,892 person-years of follow up, we documented 773 cases of MI (719 nonfatal and 54 fatal), 1907 cases of angina pectoris or coronary revascularization, and 604 confirmed cases of ischemic or hemorrhagic stroke (566 nonfatal and 38 fatal). When compared to men without prior vasectomy, the multivariate relative risk (RR) of total MI adjusted for age and other coronary risk factors was 0.94 (95% confidence interval [CI], 0.77-1.14) among men with vasectomy. Risk estimates for fatal and nonfatal events did not appreciably differ from each other. For angina or coronary revascularization or both, the multivariate relative risk was 0.99 (0.88-1.12) and for total stroke the RR was 0.95 (0.75-1.21). For men who had undergone vasectomy 15 or more years previously, the multivariate relative risks were 0.98 (0.73-1.32) for total MI, 1.17 (0.87-1.57) for total stroke, and 1.12 (0.94-1.35) for angina/revascularization. These results provide reassuring evidence that vasectomy does not materially increase the risk of subsequent cardiovascular disease, even 15 or more years following the procedure. PMID- 10382082 TI - Cycle control with oral contraceptives containing 20 micrograms of ethinyl estradiol. A multicenter, randomized comparison of levonorgestrel/ethinyl estradiol (100 micrograms/20 micrograms) and norethindrone/ethinyl estradiol (1000 micrograms/20 micrograms). AB - A randomized, open-label, multicenter study was undertaken to compare the effects of oral contraceptives (OC) containing 100 micrograms levonorgestrel (LNG)/20 micrograms ethinyl estradiol (EE) (Aless/Loette) and 1000 micrograms norethindrone acetate (NETA)/20 micrograms EE (Loestrin Fe 1/20) on menstrual cycle control over four cycles of use. A total of 84 evaluable women provided 274 cycles of exposure in the LNG/EE group, and 89 women provided 289 cycles of exposure in the NETA/EE group. Overall, the LNG/EE group achieved a consistently higher percentage of normal menstrual cycles as well as a lower rate of intermenstrual bleeding and amenorrhea than the NETA/EE group. In cycle 4, 63.8% of cycles were normal in the LNG/EE group compared with 41.9% in the NETA/EE group (p < 0.005). Of the total cycles in the NETA/EE group, 10% were amenorrheic, compared with 1.1% in the LNG/EE group. The occurrence of bleeding and/or spotting was significantly lower in cycles 2 and 3 in the LNG/EE group (41.7% and 34.8%, respectively) compared with the NETA/EE group (62.3% and 56.3%; p < 0.05). Other cycle variables were generally similar between groups, as was the incidence of adverse events. These results demonstrate that good cycle control was achieved with an OC containing 20 micrograms EE and that 100 micrograms LNG/20 micrograms EE produces better cycle control than 1000 micrograms NETA/20 micrograms EE. PMID- 10382084 TI - Normative reproductive indices for male and female adult Sprague-Dawley rats. AB - The Sprague-Dawley rat is traditionally used as the experimental model for the study of contraceptive agents and reproductive toxicants. Until recently, the normative values used to compare hormone levels after drug exposure were based on the values generated by radioimmunoassay methods developed 30 years ago. To ascertain normative reproductive indices for adult female and male Sprague-Dawley rats over a 6-month age period, we measured luteinizing hormone, testosterone, and estradiol using commercially available kits that employ updated assay techniques. In addition, sperm indices were correlated with reproductive hormones over the same time period. Animals were killed at 107, 128, 156, 212, and 268 days of age irrespective (for females) of cycle stage. Serum LH levels did not change with increasing age; however, the female rats had significantly higher LH values than did the males at comparable ages (p < 0.001). Testosterone levels and sperm parameters did not significantly change with increasing age. Estradiol levels were significantly higher in 107-day-old female rats than they were in female rats in all older age groups (p < 0.01). The values reported can be used in designing and interpreting data generated in ongoing, long term toxicological and contraceptive studies using the rat animal model. PMID- 10382083 TI - Prostate-specific antigen in vaginal fluid as a biologic marker of condom failure. AB - Forty women participated in three clinic visits during which they were exposed to their partner's semen (10 microL, 100 microL, and 1 mL). At each visit they took vaginal fluid samples before exposure to their partner's semen, immediately after, and at 1, 24, and 48 h after exposure. PSA was measured with an enzyme linked immunoassay. The mean PSA level for preexposure swabs ranged between 0.43 and 0.88 ng/mL. The mean PSA levels were 193 immediately after exposure to 10 microL, 472 after 100 microL, and 19,098 after 1 mL. The PSA levels declined within 1 h, and returned to background at 48 h. The findings confirm that our procedure is a sensitive and specific method for detecting recent semen exposure, and indicate that PSA levels depend on exposure intensity and time since exposure. Application of this method in condom efficacy studies provides objective evidence of condom failure that enhances the interpretation of self report. PMID- 10382085 TI - Costs of health care: a lopsided debate. PMID- 10382086 TI - Respective roles of scatter, attenuation, depth-dependent collimator response and finite spatial resolution in cardiac single-photon emission tomography quantitation: a Monte Carlo study. AB - The purpose of this study was to investigate the relative influence of scatter, attenuation, depth-dependent collimator response and finite spatial resolution upon the image characteristics in cardiac single-photon emission tomography (SPET). An acquisition of an anthropomorphic cardiac phantom was performed together with corresponding SPET Monte Carlo simulations. The cardiac phantom and the Monte Carlo simulations were designed so that the effect of scatter, attenuation, depth-dependent collimator response and finite spatial resolution could be studied individually and in combination. The impact of each physical effect and of combinations of effects was studied in terms of absolute and relative quantitative accuracy, spatial resolution and signal-to-noise ratio (SNR) in the resulting images. No corrections for these effects were assessed. Results obtained from Monte Carlo simulations and real acquisitions were in excellent agreement. Attenuation introduced about 90% activity underestimation in a 10-mm-thick left ventricle wall while finite spatial resolution alone introduced about 30% activity underestimation. Scatter had a negligible impact on quantitative accuracy in the recontructed slices when attenuation was present. Neither bull's eye map homogeneity nor contrast between a hot and a cold region were affected by depth-dependent collimator response or finite spatial resolution. Bull's eye map homogeneity was severely affected by attenuation but not by scatter. Attenuation and scatter reduced contrast by about 20% each. Both attenuation and scatter increased the full-width at half-maximum (FWHM) characterizing the spatial resolution of the imaging system by approximately 1 mm each but the main effect responsible for the observed 11-mm FWHM spatial resolution was the depth-dependent collimator response. SNR was reduced by a factor of approximately 2.5 because of attenuation, while scattered counts increased SNR by approximately 10%. In conclusion, the quantification of the relative influence of the different physical effects showed that attenuation is definitely the major phenomenon affecting cardiac SPET imaging accuracy, but that finite spatial resolution, scatter and depth-dependent collimator response also contribute significantly to the errors in absolute and relative quantitation and to the poor spatial resolution. PMID- 10382087 TI - An automatic classification technique for attenuation correction in positron emission tomography. AB - In this paper a clustering technique is proposed for attenuation correction (AC) in positron emission tomography (PET). The method is unsupervised and adaptive with respect to counting statistics in the transmission (TR) images. The technique allows the classification of pre- or post-injection TR images into main tissue components in terms of attenuation coefficients. The classified TR images are then forward projected to generate new TR sinograms to be used for AC in the reconstruction of the corresponding emission (EM) data. The technique has been tested on phantoms and clinical data of brain, heart and whole-body PET studies. The method allows: (a) reduction of noise propagation from TR into EM images, (b) reduction of TR scanning to a few minutes (3 min) with maintenance of the quantitative accuracy (within 6%) of longer acquisition scans (15-20 min), (c) reduction of the radiation dose to the patient, (d) performance of quantitative whole-body studies. PMID- 10382088 TI - Attenuation correction improves the detection of viable myocardium by thallium 201 cardiac tomography in patients with previous myocardial infarction and left ventricular dysfunction. AB - The aim of this study was to determine the influence of attenuation-corrected thallium-201 stress/redistribution/reinjection single-photon emission tomography (SPET) on the number of viable segments in patients with previous myocardial infarction and dysfunctional myocardium. Fifty-one patients with previous myocardial infarction and left ventricular dysfunction were included in the study. In all patients, 201Tl non-corrected (NC) and attenuation-corrected (AC) SPET was performed using a stress/redistribution/reinjection protocol followed by coronary angiography. A semiquantitative analysis was performed using polar maps for NC and AC stress, redistribution and reinjection short-axis and vertical long axis (apex) slices. Severe (perfusion defect below 50%/maximal count rate: PD < 50), mild and moderate persistent defects for redistribution and reinjection were evaluated for both NC and AC studies. A total of 1581 segments were evaluated by semiquantitative segmental analysis for both NC and AC studies for each redistribution and reinjection map. In the redistribution maps, NC revealed a total of 352 segments and AC a total of 222 segments with impaired perfusion below 50% of the maximal count rate (PD < 50). The mean number of affected segments was 6.9 +/- 5.5 in the case of NC and 4.4 +/- 4.8 in the case of AC (P < 0.001). In the reinjection maps, NC revealed a total of 263 non-viable segments (PD < 50) and AC a total of 169 non-viable segments. The mean number of affected segments was 5.2 +/- 5.3 in the case of NC and 3.3 +/- 4.2 in the case of AC (P < 0.001). Recovery of function was better predicted by AC than by NC in 20% of patients in the follow-up group. Therefore, the use of attenuation correction influences the extent of viable segments by showing more viable segments in either redistribution or reinjection maps. 201Tl imaging without attenuation correction may underestimate the extent of tissue viability, which may contribute to the lower sensitivity compared to fluorine-18-fluorodeoxyglucose positron emission tomography, where attenuation correction is a routinely performed procedure. PMID- 10382089 TI - Dobutamine stress thallium-201 single-photon emission tomography versus echocardiography for evaluation of the extent and location of coronary artery disease late after myocardial infarction. AB - Dobutamine stress echocardiography and thallium-201 myocardial perfusion scintigraphy are clinically useful methods for the evaluation of coronary artery disease (CAD). However, the relative merits of these imaging modalities in the evaluation of the extent of CAD after myocardial infarction have not been well studied. The aim of this study was to compare the accuracy of dobutamine stress echocardiography and simultaneous 201Tl single-photon emission tomography (SPET) imaging for the diagnosis and localization of CAD late after acute myocardial infarction. Dobutamine (up to 40 micrograms kg-1 min-1)-atropine (up to 1 mg) stress echocardiography in conjunction with stress-reinjection 201Tl SPET was performed for the evaluation of myocardial ischaemia in 90 patients with previous myocardial infarction who underwent coronary angiography. Significant CAD was predicted on bases of myocardial ischemia (new or worsening wall motion abnormalities on echocardiography and reversible perfusion defects on 201Tl SPET). Significant CAD (> or = 50% luminal diameter stenosis) was detected in 73 (81%) patients. The sensitivity, specificity and accuracy of echocardiography in detecting remote ischaemia for the diagnosis of remote CAD (present in 53 patients) were, respectively, 79% (CI 70%-88%), 85% (CI 77%-93%) and 81% (CI 73% 90%), while the corresponding figures for 201Tl SPET were 75% (CI 66%-85%), 78% (CI 69%-87%) and 76% (CI 67%-86%) respectively (P = NS vs echocardiography). The sensitivity, specificity and accuracy of echocardiography in detecting peri infarction ischaemia for the diagnosis of infarct-related artery stenosis (present in 70 patients) were, respectively, 77% (CI 68%-86%), 85% (CI 78%-92%) and 79% (CI 70%-87%) while the corresponding figures for 201Tl SPET were 73% (CI 64%-82%), 85% (CI 78%-92%) and 76% (CI 67%-84%) respectively (P = NS vs echocardiography). The agreement between the two methods for the diagnosis of peri-infarction and remote ischaemia was 70% (kappa = 0.37) and 80% (kappa = 0.59) respectively. It is concluded that dobutamine stress echocardiography and 201Tl SPET have comparable accuracy for the diagnosis of infarct related and remote CAD in patients with previous myocardial infarction. The agreement between the methods is higher for the diagnosis of remote CAD than for that of peri infarction ischaemia. PMID- 10382090 TI - The kinetics of beta-methyl-substituted labelled fatty acids in ischaemic myocardium: an analysis in man and with a blood-perfused isolated heart model. AB - beta-Methyl-substituted free fatty acids (FFAs) have been developed for myocardial single-photon emission tomography (SPET) imaging, but little is known about their kinetics in ischaemic conditions. The aim of this study was to determine the changes in the myocardial kinetics of a beta-methyl-branched FFA, [123I]16-iodo-3-methyl-hexadecanoic acid (MIHA), under ischaemic conditions. The kinetics of MIHA were analysed: (a) using a blood-perfused isolated heart model subjected to moderate ischaemia (50% flow reduction) and (b) in patients who had an exercise thallium-201 SPET defect corresponding to either necrotic (n = 13) or chronically ischaemic and viable (n = 15) myocardium, and who underwent two consecutive SPET studies after MIHA injection. In animals, the myocardial early retention fraction of MIHA, but not its clearance rate, was dependent on coronary flow, the early retention fraction being higher in ischaemic than in normoxic conditions (0.24 +/- 0.10 vs 0.14 +/- 0.04, P = 0.004). In the patient SPET studies, the uptake of MIHA calculated in ischaemic and viable areas (G1: 74% +/- 9% of maximal left ventricular value) was different from that calculated in necrotic (G2: 59% +/- 7%, P < 0.001) or normal (G3: 88 +/- 6%, P < 0.001) areas. By contrast, MIHA-clearance calculated between the two consecutive SPET studies was not different in G1, G2 and G3. Unlike in the case of other FFAs, the myocardial clearance of MIHA is not decreased by ischaemia. However, the early retention of MIHA is increased in the case of a moderate reduction in coronary flow, a property which might help in the detection of viability in chronically ischaemic myocardium. PMID- 10382091 TI - Normal appearances of technetium-99m dimercaptosuccinic acid in children on planar imaging. AB - Thirty-two children who underwent renal arteriography with normal results also underwent technetium-99m dimercaptosuccinic acid (DMSA) imaging. Variations in the normal appearance of 99mTc-DMSA images are described in these children. Criteria for high-quality 99mTc DMSA images are defined in terms of kidney outline and contrast between the inner and the outer part of the kidney. Most of the images in this study were of high quality. The two most common features of normal 99mTc-DMSA images were a round-shaped (50%) or flat (either lateral or medial aspect) (24.8%) contour. A small number of unusual appearances were observed and these have been illustrated. The mean differential function of the left kidney was 51% with a range of 45%-57%. PMID- 10382092 TI - Rapid renal single-photon emission tomography by continuous infusion of iridium 191m. AB - Continuous infusion of iridium-191m (t1/2 = 5 s), produced with an 191Os/191mIr generator, was used to obtain rapid high-resolution single-photon emission tomography (SPET) of renal blood flow in the rabbit. SPET scans of the abdomen were obtained with a triple-detector SPET system (MS3, Siemens Gammasonics). The generator was eluted at a flow rate of 3 ml/min, which delivered a steady-state dose of 170 MBq (4.5 mCi) of 191mIr. The total 191Os breakthrough was 850 kBq (23 microCi). A 5-min SPET acquisition recorded a total of 2.8 million counts, resulting in images of high technical quality. Volume-rendered images clearly showed the abdominal aorta, splenic artery, spleen, renal arteries, kidneys and splanchnic vasculature. Tomographic slices through the kidneys revealed tracer primarily within the renal cortices without visualization of the collecting system. The estimated effective dose equivalent for a 5 min infusion of 191mIr at a steady-state dose of 170 MBq is 0.74 mSv compared with 2.7 mSv from a 170 MBq dose of 99mTc-DMSA. This study demonstrates the feasibility of high-resolution SPET of regional renal perfusion in the rabbit by continuous intravenous infusion of 191mIr. The renal distribution of continuously infused 191mIr is largely within the cortices, with minimal or no detectable activity in the region of the renal pelvicalyceal system. Using this technique, cortical renal SPET can be completed much more rapidly (< 5 min) than with conventional renal cortical imaging agents, which suggests that this technique could be applied to the observation of rapid changes in renal perfusion such as those resulting from pharmacologic intervention, obviating the need for the patient to return for additional visits. Additional studies are required to (a) validate the methodology in larger animals prior to considering the potential for use in human beings, (b) optimize the generator design for continuous infusion, and (c) evaluate the changes in the distribution of 191mIr that occur in animal models of altered renal perfusion. PMID- 10382093 TI - Evaluation in mice of some iodinated melanoma imaging agents using cryosectioning and multi-wire proportional counting. AB - AMBIS 4000, a multi-wire proportional counter, was calibrated for iodine-125 measurements. The detector displayed a linear response over a wide dynamic range. Using whole-body mice cryosections, a linear relationship could be established between count rate per area (cpm/mm2) measured with the AMBIS 4000 detector and the count rate per gram (dpm/g) determined with an NaI(Tl) scintillation detector. A calibration curve could, thus be constructed. This new method allowed direct visual and quantitative evaluation of the biodistribution of a short series of 125I-labelled benzamides in melanoma-bearing mice. All the compounds studied showed good tumoral targeting ability. For one of them, ortho-N-(2 diethylaminoethyl)-4-iodobenzamide, liver and lung uptake decreased rapidly after dosing, making it a suitable tracer for scintigraphic detection of malignant melanoma. PMID- 10382094 TI - Preoperative assessment of cervical lymph nodes in head and neck cancer with fluorine-18 fluorodeoxyglucose using a dual-head coincidence camera: a pilot study. AB - The aim of this study was to investigate whether in patients with head and neck cancer, staging is possible with fluorine-18 fluorodeoxyglucose (18F-FDG) using a dual-head positron emission tomography (PET) camera. Twenty patients (ten men, ten women; mean age: 60 years) were studied using 185 MBq (5 mCi) 18F-FDG. Two of these patients who were suspected of having recurrence in the neck were restaged 19 and 12 months, respectively, after the resection of the primary tumour. The images were visually analyzed and the results were correlated with computed tomography (CT) (n = 18), ultrasonography (n = 17) and pathological findings. With respect to the primary tumour, FDG dual-head PET and CT revealed a sensitivity of 100% and 59%, respectively (P < 0.001). In seven patients lymph node metastases were found in the neck specimen. Two of them had bilateral metastases. FDG dual-head PET correctly identified all nine pathological neck sides whereas CT and ultrasonography depicted eight of nine and seven of eight pathological sides, respectively. In three patients, false-positive FDG uptake was seen, which was due to a preceding biopsy in two cases. The sensitivity of FDG dual-head PET, CT and ultrasonography in the identification of pathological neck sides was 100%, 89% and 87%, respectively, and the specificity was 90%, 93% and 50%, respectively. With knowledge of the preceding biopsies, the specificity of FDG dual-head PET would have been 97%. The smallest lymph node metastasis detected by FDG dual-head PET that was missed by CT had a diameter of 0.6 cm. Measurement of 18F-FDG with a dual-head PET camera is very sensitive in the detection of primary head and neck cancers and accurate in the preoperative assessment of lymph node metastases. The results justify a prospective study on the identification of metastases in patients with head and neck cancer. In addition, it is justified to start a study on the detection of unknown primary tumours in patients with cervical metastases. PMID- 10382095 TI - Measurement of lymphatic function with technetium-99m-labelled polyclonal immunoglobulin. AB - A reliable method for measuring lymph flow in physiological units would be valuable, especially in conditions in which it is uncertain whether lymph flow is increased or decreased. The requirements of a radiopharmaceutical for such measurement include stable radionuclide labelling and rapid access to lymphatic vessels following tissue injection but no access to blood vessels. A soluble macromolecule is likely to come closest to meeting these requirements. Technetium 99m-labelled human polyclonal immunoglobulin (HIG) was therefore investigated firstly in comparison with 99mTc-labelled human serum albumin (HSA) in patients undergoing routine lymphoscintigraphy and secondly with respect to injection site in a group of volunteers with post-mastectomy oedema (PMO). Subcutaneous injection of 99mTc-HIG into the web space of a distal extremity gave images in which lymphatic vessels were more clearly defined compared with images obtained after injection of 99mTc-HSA. Lymph nodes were also more clearly defined, suggesting specific retention of HIG, possibly through Fc-mediated binding. Peripheral blood sampling showed a delayed arrival in blood of radioactivity after 99mTc-HIG compared with 99mTc-HSA, although ultimately, the blood recovery of 99mTc-HIG was significantly higher (P < 0.05) than that of 99mTc-HSA. Clearance rates of radioactivity from the injection site were not significantly different, however, between the two agents. In patients with PMO, web space injection of 99mTc-HIG gave excellent images of normal lymphatic vessels, of lymph nodes and of abnormal lymph drainage such as dermal backflow in swollen arms. In contrast, neither lymphatic vessels nor lymph nodes were visualised after injection into the skin of the dorsum of the distal forearm. Although there was no difference in clearance rates from the injection sites between normal and swollen arms with either agent in PMO, clearance was significantly faster following injection into the web space (0.11% per minute for normal and swollen arms combined) than into the forearm (0.053% per minute; P < 0.05). These results suggest that (a) 99mTc-HIG is a potentially useful agent for measuring lymph flow and lymph node function; but (b) injection into the dorsum of the forearm is not a useful method of administration for these measurements; and (c) clearance rates from the injection site do not support the notion that PMO is the result of decreased lymph flow. Further studies are warranted to evaluate 99mTc-HIG as an agent for assessment of lymphatic function, especially with respect to measurement of lymph flow and possibly also for the evaluation of lymph node Fc mediated immunocompetence. PMID- 10382096 TI - Somatostatin receptor scintigraphy to predict the clinical evolution and therapeutic response of thyroid-associated ophthalmopathy. AB - Management of thyroid-associated ophthalmopathy remains a topic of controversy. Immunosuppressive treatments have to be applied at peak disease activity and before criteria of severity develop. Expression of somatostatin receptors on activated lymphocytes allows scintigraphic imaging with indium-111 pentetreotide. We conducted a prospective study with 17 patients who presented severe ophthalmopathy (11 Graves' disease, four Hashimoto's thyroiditis, two isolated in appearance: Means' syndrome). Each patient underwent hormonal (free T3 and TSH) and immunological (TBII) assessment, an orbital computed tomography scan or magnetic resonance imaging, a visual functional examination and 111In pentetreotide orbital scintigraphy before undergoing treatment by steroids and/or radiotherapy, independently of scintigraphic results. At 4 and 24 h after the intravenous injection of 111 MBq of 111In-pentetreotide, planar imaging centred on the head and neck (anterior and both lateral views) was carried out. Retrobulbar uptake was assessed by visual semi-quantitative analysis (score given by two independent trained observers) and by quantitative analyses (regions of interest, orbit/brain uptake indices). Patients were ophthalmologically followed up for 6 months and then classified as improved or not. Visual semi-quantitative analysis of 4-h/24-h planar images was correlated with the ophthalmological evolution (chi 2 test, P < 0.01). All ten patients in whom scintigraphy was considered positive were clinically improved at 6 months, and of the seven patients in whom scintigraphy was negative, six were not improved. Nevertheless, objective quantitative analysis did not succeed in confirming these results. We conclude that 111In-pentetreotide scintigraphy requires further developments, including quantitative single-photon emission tomographic acquisition, if its role as a guide to therapeutic strategy in thyroid-associated ophthalmopathy is to be confirmed. PMID- 10382097 TI - Simplified reference region model for the kinetic analysis of [99mTc]TRODAT-1 binding to dopamine transporters in nonhuman primates using single-photon emission tomography. AB - Accurate quantification of neuroreceptors requires full kinetic modeling of the dynamic single-photon emission tomography (SPET) or positron emission tomography (PET) images, with highly invasive arterial blood sampling. This study investigated the application of a reference region kinetic model to the dynamics of [99mTc]TRODAT-1 in nonhuman primates, obviating the need for blood sampling. A series of dynamic SPET scans were performed on two baboons following the injection of approximately 700 MBq of [99mTc]TRODAT-1. Rapid arterial blood samples were taken automatically during scanning. Reconstructed SPET images were coregistered with magnetic resonance imaging (MRI) scans of the baboons, and regions of interest (ROIs) placed on the striatum, cerebellum and cerebral hemispheres. The ROI data were combined with metabolite-corrected blood data, and fitted to a three-compartment kinetic model using nonlinear least squares techniques. The same data were also used in a simplified reference region model, in which the input function was derived from the nondisplaceable tissue compartment. In addition, semiquantitative blinded analysis was performed by three raters to determine the point of transient equilibrium in the specific binding curves. All methods generated values for the ratio of the kinetic rate constants k3/k4, which gives an estimate of the binding potential, BP. These were compared with the full kinetic model. The mean values of k3/k4 for the three different analysis techniques for each baboon were: 1.17 +/- 0.21 and 1.12 +/- 0.13 (full kinetic model), 0.93 +/- 0.13 and 0.90 +/- 0.07 (reference region model), and 0.97 +/- 0.18 and 0.92 +/- 0.08 (equilibrium method). The reference region method gave significantly lower results than the full kinetic model (P = 0.01), but it also produced a much smaller spread and better quality fits to the kinetic data. The reference region model results for k3/k4 correlated very strongly with the full kinetic analysis (r2 = 0.992, P < 0.001), and with the equilibrium model (r2 = 0.88, P = 0.002). The subjectivity inherent in the equilibrium method produces inferior results compared with both kinetic analyses. It is suggested that the self-consistency of the reference region model, which requires no arterial blood sampling, provides a more precise and reliable estimate of the binding of [99mTc]TRODAT-1 to dopamine transporters than full kinetic modeling. The reference region method is also better suited to a routine clinical environment, and would be able to distinguish smaller differences in dopaminergic function between patient groups. PMID- 10382098 TI - Recommendations for implementing SPECT instrumentation quality control. Nuclear Medicine Section--National Electrical Manufacturers Association (NEMA). PMID- 10382099 TI - Nuclear medicine procedures in lung cancer. AB - Although radiography, computed tomography and magnetic resonance imaging are still the methods of choice for the study of lung cancer, they have certain limitations in the determination of the nature of suspicious lung nodules, the evaluation of mediastinal involvement, the assessment of the viability of previously treated lesions and the diagnosis of tumour relapse. There is a wide range of current oncological requirements related to lung cancer: detection of malignant lesions at the earliest stage and in the most effective way; the definition of the biological characteristics of a lesion (proliferation, aggressiveness, differentiation, etc.); the need to define the operability of the patient (function of residual lung and staging); and the need to evaluate the behaviour of the tumour (response to therapy, early detection of recurrences, metastatic spread). Most of the efforts of the nuclear medicine community have been focussed on diagnosis, staging, restaging and therapy monitoring of lung cancer. Many radiopharmaceuticals have been employed for this, including gallium, monoclonal antibodies, somatostatin analogues, lipophilic cations and positron emission tracers. There is ample evidence that nuclear medicine techniques may provide complementary information with respect to anatomical imaging, for example in the assessment of preoperative function by means of ventilation and perfusion scintigraphy, or in tumour localisation by means of specific tumour-seeking agents. However, clinical data suggest that, when properly used, nuclear medicine procedures in some cases may be not only complementary to radiology but essential for the clinical management of lung cancer. An example of such a procedure is fluorodeoxyglucose positron emission tomography (FDG PET) the introduction of which has greatly contributed to confirmation of the clinical value of nuclear medicine in this field. FDG PET has proved of great help in lung cancer management and its cost-effectiveness in lung cancer staging is firmly established. In this review the results of the most important nuclear medicine techniques are summarised and their value in clinical practice is discussed. General, updated information is provided about the epidemiology, biology and clinical management of lung cancer, and about the role of nuclear medicine in these areas. PMID- 10382100 TI - Scientific poster presentation at the EANM-meeting: a penalty system would solve the problem immediately. PMID- 10382102 TI - 1999: main diagnostic applications of nuclear medicine in oncology. Task Group of Oncology, World Federation of Nuclear Medicine and Biology. PMID- 10382101 TI - Current status of commercial colloidal preparations for sentinel lymph node detection. PMID- 10382103 TI - Diphtheria and tetanus toxoid microencapsulation into conventional and end-group alkylated PLA/PLGAs. AB - The feasibility of biodegradable polyester microspheres (MS) for single injection vaccines will greatly depend on the toxoid stability within the MS exposed to in vivo conditions. This study examined the effects of polymer type and co encapsulated additives on diphtheria (Dtxd) and tetanus (Ttxd) toxoid entrapment and stability. The co-encapsulated stabilizers influenced significantly the entrapment of Dtxd and Ttxd in PLA/PLGA MS. Typically, 5% BSA or trehalose decreased the amount of Dtxd entrapped in spray-dried MS, whereas BSA increased the entrapment in coacervated MS. Further, the entrapment of Dtxd decreased as a function of polymer hydrophobicity in spray-dried MS. Without additives, approx. 64, 43 and 16% entrapment efficiency of ELISA-reactive antigen was obtained for 14-17 kDa PLGA 50:50, PLGA 75:25 and PLA, respectively. The novel end-group stearylated 1-PLAs were only processed by coacervation. Satisfactory entrapment of 30-60% Dtxd was obtained. Here, albumin was a prerequisite for toxoid encapsulation, as BSA-free formulations produced strong toxoid precipitation. Furthermore, protein burst release increased with the more hydrophobic polymers, with Dtxd, Ttxd and the co-encapsulated BSA following a similar pattern and magnitude. This investigation also revealed that the method of protein extraction from the microspheres (O/W-partition or polymer hydrolysis) as well as the analytical methods (HPLC or ELISA) strongly influenced the determined amount of encapsulated toxoid and BSA. In conclusion, the study revealed the complexity of antigen microencapsulation when using different preparation and analytical techniques, as well as different types of materials. PMID- 10382104 TI - Evaluation of aminoalkylmethacrylate nanoparticles as colloidal drug carrier systems. Part I: Synthesis of monomers, dependence of the physical properties on the polymerization methods. AB - Conventional nanoparticles based on acrylic compounds are lipophilic and possess a negative surface charge. This is due to their manufacturing process and to the chemical structure of the polymer. Hence, these particles are not suitable for the adsorption of hydrophilic anionic drugs. In the present investigation, positively charged copolymer nanoparticles prepared from aminoalkyl- and methylmethacrylates were evaluated, with regard to their physical properties. This report provides a detailed description of the synthesis of the non commercially available monomers and their polymerization procedure. Various parameters were investigated, such as comonomer content, total amount of monomer, concentration of the radical initiator, and the composition of the polymerization medium. The resulting particle diameter and the surface charge were found to be strongly dependent on the polymerization conditions and on the pH. Optimization of the polymerization procedure yielded nanoparticles of about 200 nm exhibiting a positive surface charge. The charges of the different copolymer particles were then compared at different pH values. N-trimethylaminoethylmethacrylate (TMAEMC) nanoparticles with quaternary ammonium groups located at their surfaces, possessed a nearly constant positive zeta potential at various pH values and, consequently, pH-independent particle diameters. The physical characteristics of the other aminoalkyl copolymers correlated with the basicity of the monomers employed and were found to be strongly dependent on the pH of the dispersion medium. Aminoethylmethacrylate (AEMC), methylaminoethylmethacrylate (MMAEMC), and aminohexylmethacrylate (AHMC) as well as aminoethylmethacrylamide (AHMAC) copolymer nanoparticles exhibited a strong positively charged surface even at physiological pH and, therefore, are useful candidates for the adsorption of anionic drugs. PMID- 10382105 TI - The effect of formulation parameters on the size of poly-((2-dimethylamino)ethyl methacrylate)-plasmid complexes. AB - The aim of this study was to gain insight into the formulation parameters affecting the size of poly((2-dimethylamino)ethyl methacrylate)-plasmid complexes (polyplexes). Experimental designs were applied to screen and optimize several variables, which may influence the complex size. In a screening design, it was demonstrated that at a fixed concentration of plasmid (40 micrograms/ml) after incubation with polymer, the size of the resulting polyplexes was highly dependent on the polymer/plasmid ratio as well as on the pH, viscosity (i.e. sucrose concentration) and ionic strength of the aqueous solution. However, the temperature, PEG 600 (up to 5% (v/v)) and Tween 80 (up to 0.2%) had a marginal effect on the size of the polyplexes. In an optimization design, the effect of the pH, polymer/plasmid ratio and Tween on the size of the polymer/plasmid complexes prepared at relatively high concentration of plasmid (50-200 micrograms/ml) was evaluated. Based on the results of the optimization design, a mathematical model was derived, which describes the relationship between the size of the polyplexes and the different formulation parameters. This model shows that even at high plasmid concentration (200 micrograms/ml), small sized polyplexes were formed at low pH and ionic strength, especially when the solution contains 20% (w/v) sucrose. This concentrated polyplex dispersion (polymer/plasmid ratio > 3/1 (w/w), 200 micrograms plasmid/ml) can be diluted down to 5 micrograms/ml plasmid without significant changes in particle size and transfection potential. At lower ratios, a growth in particle size was observed upon dilution of the complexes, which might also explain the low transfection efficiency of these polyplexes in vitro. PMID- 10382106 TI - Effect of crystallinity of microcrystalline cellulose on the compactability and dissolution of tablets. AB - Microcrystalline cellulose (MCC) was pulverized with a vibrational rod mill. The degree of crystallinity of MCC decreased from 65.5 to 12.1% with pulverization time due to mechanochemical effect. Pulverized MCCs were compressed at 155.6 MPa using a compression test apparatus, and the two parameters relating to compactability, the B value and yield pressure, were calculated using a Heckel plot. These values were lowered as the degree of crystallinity of MCC became smaller. These results suggest that the crystal region and the amorphous region in MCC particles may be mainly fractured and deformed plastically during compression, respectively. Then the dissolution test was performed for the acetaminophen-MCC (10:90) tablets. Dissolution profiles showed an interesting phenomenon, namely, the dissolution rate of acetaminophen from MCC tablet decreased when the degree of crystallinity of MCC was in the range from 65.5 to 37.6%, however, it increased markedly when the degree of crystallinity of MCC was in the range from 25.8 to 12.1%. The amount of water absorbed into tablets changed in accord with the dissolution rates of acetaminophen from tablets. The dissolution data indicate that drug release can be modified by changing the degree of crystallinity of MCC. PMID- 10382107 TI - The tabletting machine as an analytical instrument: qualification of the tabletting machine and the instrumentation with respect to the determination of punch separation and validation of the calibration procedures. AB - The quality of the determination of punch separation in an eccentric tabletting machine equipped with two inductive displacement transducers was carefully investigated, since this tabletting machine is used as an 'analytical instrument' for the evaluation of the compression behaviour of pharmaceutical materials. For a quasistatic calibration procedure using gauge blocks, the repeatability under standard conditions and the robustness against variations in machine settings, installation conditions, equipment and methods were examined. The readings during calibration can be easily influenced by machine parameters as a result of deficiencies in the construction of the machine and in the mode of instrumentation. The poor plane-parallelism of the punch faces has a further negative effect on the accuracy of punch separation. In addition, the response at loading to lower and higher forces as during calibration was investigated. While at loading up to 100 N, the response of the system to the gauge blocks is systematically influenced by punch separation, for slow manually applied punch-to punch loading up to 16.5 kN at a broad range of penetration depths, no significant effects were observed in the region of interest for tabletting. To get an indication of the transferability of the calibration and the determination of punch deformation to normal operating conditions, the lateral tilting of the punches during dynamic idle runs, punch-to-punch loading, and compression of microcrystalline cellulose was analyzed. A transfer of the response derived from punch-to-punch compression to tabletting conditions seems to be possible, although this must be questioned on grounds of theoretical considerations. From all the experiments performed, a total error of about +/- 30 microns must be assessed for the determination of punch separation. PMID- 10382108 TI - Radiolabelling of parenteral O/W emulsions by means of neutron activation. AB - Parenteral O/W emulsions containing lanthanide fatty acid derivatives were prepared. With regard to enhancing the incorporation efficiency of the neutron activatable excipients, the addition of the non-ionic co-emulsifier Solutol HS 15 proved to be most suitable. Comparing the different chain lengths of the fatty acids, the long chain fatty acid derivative lanthanide(tri)stearate seemed to be superior in strengthening the interfacial layer. After neutron activation, the physical and chemical stability of the irradiated formulations was evaluated. The chemical stability, indicated by the concentration of lyso phosphatidylcholine as the degradation product of the main emulsifier, was shown to be dependent on the irradiation time. By applying a neutron flux of 2.1 x 10(13) neutrons/cm2 per s, the maximum should not rise above 60 s. The physical stability indicated by the particle size distribution was affected by the presence of the non-ionic co emulsifier. Concerning the amount of radiation necessary for in vivo biodistribution studies the maximum load of Samarium fatty acid derivatives did not yield sufficient radioactivity levels. However, Europium derivatives could be shown to be suitable for in vivo studies. PMID- 10382109 TI - Studies on the effect of pilocarpine incorporation into a submicron emulsion on the stability of the drug and the vehicle. AB - In order to obtain a novel ocular formulation with a potential for prolonging pilocarpine activity, the drug (2.0%) was incorporated into a submicron emulsion containing soya-bean oil and lecithin as emulgator. The effect of drug incorporation into the emulsion on its physical stability and on the other hand, the potential of the vehicle to reduce drug degradation at pH higher than 5.0 was studied. The pH was adjusted to 6.5 or 5.0 and the physicochemical stability of the formulations was observed. The mean diameter of oily particles in the resulting emulsions measured by a laser diffractometer was 0.6-0.7 micron and this was larger than in a drug-free emulsion where a 0.33 micron value was measured. The formulations were physically stable for 6 months at 4 degrees C, but progressing chemical degradation of pilocarpine was noted at pH 6.5. At that pH nearly 8% of pilocarpine was degraded to isopilocarpine and pilocarpic acid, both in the emulsion and in the solution. Thus, it may be concluded that pilocarpine in submicron emulsion is not protected against degradation. The presence of pilocarpine changes the physical stability of the vehicle since the formulation was easily destabilized during autoclaving or at room temperature. In the presence of higher concentration of lecithin (2.4%) or co-emulgators (poloxamer 2.0% or Tween 80 0.5%) the mean droplet size in the emulsions was the same as in a drug-free system. However the emulsions containing poloxamer were not stable during storage. Viscosity of pilocarpine emulsions can be increased by addition of methylcellulose or sodium carmellose (1.0%), but an intensive creaming occurs in these systems. Pilocarpine base is less suitable for emulsion preparation than hydrochloride salt, and emulsions prepared at pH 5.0 show the most satisfying stability. PMID- 10382110 TI - Analysis of chromameter results obtained from corticosteroid-induced skin blanching assay: comparison of visual and chromameter data. AB - In a Guidance document, the American FDA recommends the use of a Minolta chromameter rather than the human eye for the quantitative assessment of the pharmacodynamic blanching response produced by topical application of corticosteroids. The purpose of this study was to compare the appropriateness of the human eye and two models of chromameter for the estimation of skin blanching, in terms of the quality of the data generated by each method. The corticosteroid induced skin blanching from four different betamethasone 17-valerate cream formulations was compared in a typical human skin blanching trial. The optimized assay methodology routinely practised in our laboratories was utilized. The blanching responses were assessed visually by three trained, independent observers and recorded by two chromameters (Minolta model CR-200 and model CR 300). The topical availability of the four creams was determined using visual scoring and chromameter measurements. All data were manipulated in such a manner as to produce a blanching response versus time profile from which AUBC analysis could be performed. Good correlation was observed between the visual assessments made by three independent observers. In contrast, moderate correlation was determined between visual, CR-200 and CR-300 measurements. Surprisingly, no direct linear relationship between the AUBCs produced by the two chromameters was observed indicating that the quality of the data obtained from the two instruments may not be equal. This investigation also indicated that the use of the chromameter is not completely objective. Visual scoring and chromameter measurement produce data sets that differ in quality. Each procedure needs to be validated and investigators have to be trained for both visual assessment and the operation of the chromameter, particularly with regard to the manipulation of the measuring head of the instrument. PMID- 10382111 TI - Effect of the degree of quaternization of N-trimethyl chitosan chloride on the permeability of intestinal epithelial cells (Caco-2). AB - N-trimethyl chitosan chloride (TMC), a partially quaternized derivative of chitosan with superior water solubility, was synthesized with different degrees of quaternization [12.6% quaternized (TMC-L) and 19.9% quaternized (TMC-H)] and the effects of these novel polymers on the permeability of intestinal epithelial cells were investigated in Caco-2 cell monolayers. Transepithelial electrical resistance (TEER) measurements showed that both polymers in 1.5-2.5% w/v concentrations caused a pronounced, concentration dependent lowering in TEER values, but that TMC-H was more effective than TMC-L at similar concentrations (36 +/- 3% reduction with TMC-L and 53 +/- 6% reduction with TMC-H at 2.0% concentrations). Similar results were obtained in transport studies with the hydrophilic radioactive markers [14C]mannitol (MW 182.2) and [14C]polyethylene glycol 4000 ([14C]PEG 4000, MW 4000). The transport of [14C]mannitol was increased 51-fold (TMC-L) and 97-fold (TMC-H) at 2.5% concentrations. No deleterious effects to the cells could be demonstrated with trypan blue exclusion studies. The results show that TMC is able to open the tight junctions of intestinal epithelial cells to allow for paracellular transport of hydrophilic molecules. It is concluded that the charge density of TMC, as determined by the degree of quaternization, is an important factor determining its potential use as an absorption enhancer across intestinal epithelia. PMID- 10382112 TI - In vitro biodegradation study of acetyl and methyl inulins by Bifidobacteria and inulinase. AB - The purpose of this study was to investigate if acetylated and methylated inulins can be degraded by inulinase from Aspergillus niger and by Bifidobacteria, in order to determine their potential in colonic drug delivery. Methyl and acetyl inulins were synthesized by the reaction of inulin (Raftiline HP) with methyl sulfate and acetic anhydride, respectively. The degree of substitution (DS) and the structure of the reaction products were confirmed by 13C-NMR. Degradation by inulinase was investigated in a citrate buffer (0.05 M, pH 4) or with a mixture of citrate buffer and DMSO at 37 degrees C. Biodegradation by Bifidobacteria was investigated under anaerobic conditions using an in-house prepared broth at 37 degrees C for 2 days. The resulting products were analyzed chromatographically; the formation of short chain fatty acids was followed by measuring the pH of the incubation media. The results obtained suggest that acetylated and methylated inulins are not biodegradable, since no degradation product could be detected after incubation; a decrease in pH was clearly observed in control samples (pure fructose and inulin), but not in the derivatized inulin samples. The results can probably be explained by a change in conformation of inulin due to derivatization. PMID- 10382113 TI - Utilizing vehicle imbibition by a microporous membrane and vehicle viscosity to control release rate of salbutamol. AB - The possibility to control the release rate of salbutamol through the hydrophobic Celgard 2500 polypropylene membrane by varying the composition and the viscosity of hydrophilic drug vehicles was investigated. The use of the polypropylene membrane as a control membrane for reservoir-type drug delivery systems was envisaged and water, glycerol, isopropyl alcohol and ethanol, pure or as binary mixtures were studied as vehicles. With varying composition of the vehicle, a sharp change of its imbibition by the membrane from practically none to a complete filling of the membrane pores occurred, which coincided with a steep rise of the drug permeability for the membrane. From this was inferred that the vehicle-filled pores were the dominant permeation pathway, while when no vehicle was imbibed, transport took place by way of the polymer domain of the membrane. In case of imbibition, the permeation rate could be modulated in a predictable fashion by adjusting the viscosity of the vehicle. This demonstrated that drug release could be controlled by utilizing the in situ interaction of the vehicles with this membrane, leading to imbibition and establishment of a permeation pathway with pre-determined viscosity in the pores of the membrane. PMID- 10382114 TI - Solubility behaviour of haloperidol in individual solvents determination of partial solubility parameters. AB - The solubility behaviour of haloperidol in individual solvents ranging from non polar to highly polar solvents was studied. Extended Hansen's method was used to analyze the solubility data and obtain partial solubility parameters of haloperidol. Flory-Huggin's size connection term 'B' was found to further improve the prediction of solubility. A four parameter extended Hansen's approach involving proton-donor and proton-acceptor parameters was also used in fitting the solubility data to a theoretical model. The term Wh, used as an empirical measure of solute-solvent interaction due to hydrogen bonding was used in calculating B. Different approaches were thus used in fitting the experimental solubility data to obtain regression equations which aim to provide a reasonable prediction of solubility of haloperidol in untested solvents. Solubility parameter was calculated from the partial solubility parameter values obtained from the different methods of data analysis, and compared with the theoretically obtained values. Solubility parameter of haloperidol is fixed at 10.58 H. PMID- 10382115 TI - Temperature effect on serum protein binding kinetics of phenytoin in monotherapy patients with epilepsy. AB - The effects of temperature on the binding kinetics of phenytoin (PHT) to serum proteins were determined in patients with epilepsy. Serum samples examined in the study were obtained from 59 patients (31 male, 28 female) with epilepsy on PHT monotherapy. Their age ranged from 3 to 64 years (mean (SD), 23.3 (16.3) years). Protein binding of PHT was evaluated by ultrafiltration under current routine laboratory conditions (25 +/- 3 degrees C) or at a temperature of 37 degrees C. The in vivo binding parameters of PHT to serum proteins were determined using a binding equation derived from the Scatchard equation for a one-site binding model. Significant differences were observed in serum concentrations of unbound PHT between paired data (P < 0.05). The mean association constant (K) of PHT to serum proteins is 0.011 microM-1 at 25 +/- 3 degrees C and 0.006 microM-1 at 37 degrees C, while mean total concentration of binding sites (n(Pt)) is 1002 microM for 25 +/- 3 degrees C and 1112 microM for 37 degrees C. Significant differences were observed in the binding kinetics of PHT to serum proteins for the different temperature conditions of ultrafiltration (P < 0.05). Our study confirms that binding affinity for PHT-serum protein interaction is approximately 45% lower at 37 degrees C than at 25 +/- 3 degrees C and consequently, binding potential (K.n(Pt)) is approximately 39% lower at 37 degrees C than at 25 +/- 3 degrees C. PMID- 10382117 TI - Concentration at steady-state after periodic and non-uniform administration of drug. AB - A two-compartment model, with absorption from the gut and elimination from both compartments, is considered in order to express the concentration at any time and at steady-state when a drug is administered repeatedly according to a dosing schedule non-uniformly distributed over a 24-h interval. A time-delay is included to take into account the drug crossing from the gut to the central compartment. PMID- 10382116 TI - Influence of pharmaceutical gel vehicles containing oleic acid/sodium oleate combinations on hairless mouse skin, a histological evaluation. AB - Aqueous gel preparations containing oleic acid/sodium oleate combinations were applied three times daily to hairless mice (CD1 strain). Six groups of animals (n = 9 or n = 10) were treated topically with six experimental vehicles containing 2, 3 and 4.5% oleic acid (OA) at two different pH values, 7.3 and 7.7. Sodium lauryl sulfate (5%) in a similar gel preparation was used as the positive control (n = 5), while untreated animals were used as the negative control (n = 6). After three treatment days, the skin samples were collected and processed for histological evaluation. It was seen that the severity and frequency of histological changes in the skin treated with OA-containing vehicles were directly correlated with increased pH/ionization (i.e. decreased OA/sodium oleate ratio) and with overall OA concentration. PMID- 10382118 TI - Helicobacter pylori and gastric cancer: what are the benefits of screening only for the CagA phenotype of H. pylori? AB - BACKGROUND: Strains of Helicobacter pylori that express the CagA protein are associated with a threefold increased gastric cancer risk as compared to H. pylori strains that do not express CagA. Screening and treatment only for CagA antibodies should target those individuals at highest gastric cancer risk while reducing the number of patients requiring antibiotics. We compared the costs and benefits of screening asymptomatic 50-year-old individuals for CagA, screening for all H. pylori strains, and no screening, both in the United States and abroad. MATERIALS AND METHODS: We employed Markov cost-effectiveness analysis using data from randomized, case-control, and cohort studies. RESULTS: In the United States, CagA screening would result in 1.5 million fewer antibiotic treatments but would prevent 1,400 fewer gastric cancers than would screening for all H. pylori. The incremental cost-effectiveness of CagA screening is $23,900 per life-year gained; for H. pylori screening, it is $25,100. Screening in countries with epidemiological characteristics similar to those of Colombia, Finland, and Japan costs less than $5,000 per life-year gained, and the difference between CagA and H. pylori screening is smaller than that in the United States. CONCLUSIONS: Screening only for CagA-positive H. pylori is not substantially better than is screening for all H. pylori, either in the United States nor abroad. Screening is substantially more cost-effective outside the United States. Whether population screening is justified, however, is uncertain pending conclusive data regarding the reduction in gastric cancer risk from antibiotics. PMID- 10382119 TI - Invasiveness of Helicobacter pylori into human gastric mucosa. AB - BACKGROUND: Helicobacter pylori has generally been observed only in the gastric mucous layer or in the spaces between gastric mucus-secreting cells and not in the gastric epithelial cells or in the lamina propria. The purpose of this study is to determine whether H. pylori invades the gastric mucosa, using an immunoelectron microscopical examination of human gastric mucosa infected with H. pylori. MATERIALS AND METHODS: Five hundred gastric antral biopsy specimens were fixed in a periodate-lysin-paraformaldehyde solution, embedded in Lowicryl, sectioned, and examined with a light microscope. One hundred specimens moderately or severely infected with H. pylori were selected and were incubated with polyclonal rabbit anti-H. pylori antibody. The specimens were washed, incubated with 20 nm of colloidal gold-conjugated goat anti-rabbit IgG, stained with uranyl acetate and lead citrate, and observed with a transmission electron microscope. RESULTS: In one case, a bacterium was observed within the cytoplasm of a gastric mucus-secreting cell; in another case, a few bacteria were observed within the cytoplasm of a stromal cell in the lamina propria. The bacteria could be differentiated from degenerated intracellular organelles by gold particles attached to the bacteria. CONCLUSION: H. pylori rarely invade the lamina propria and gastric cells. PMID- 10382120 TI - Differences among Helicobacter pylori strains isolated from three different populations and demonstrated by restriction enzyme analysis of an internal fragment of the conserved gene hpaA. AB - BACKGROUND: Our goal was to test the idea that Helicobacter pylori genotypes vary from one population to another. METHODS: Analysis of Sau3A and HinfI restriction fragment-length polymorphism (RFLP) in a 375-bp polymerase chain reaction amplicon of hpaA was used to compare 31 H. pylori isolates from a relatively small and genetically homogeneous population (Goteborg, Sweden) with those of large, genetically heterogeneous populations located in two different countries (50 isolates from Houston, TX, and 69 isolates from Minas Gerais, a state in the southeastern region of Brazil). RESULTS: Five different Sau3A and three different HinfI restriction patterns were found; different combinations of these comprise 10 different RFLP types, I through X. The RFLP types found in the United States and Brazil collections were very similar, except for two Brazil isolates belonging to type VIII and five Brazil isolates belonging to type X, neither type found in the United States. The overall profile of H. pylori isolates from Sweden was remarkably different, with 18 of 31 (58%) having a new Sau3A restriction pattern, termed gS; 10 of these 18 isolates had HinfI restriction pattern E (RFLP type VIII), and 8 had HinfI restriction pattern F (RFLP type IX). No isolates from Sweden belonged to RFLP type III or type X. CONCLUSIONS: RFLP typing of a 375-bp polymerase chain reaction-amplified DNA fragment of H. pylori hpaA revealed that H. pylori genotypes can and do vary from one population to another. We conclude that the unique RFLP profile shown by the group of H. pylori isolates from Goteborg is the result of a cohort effect in this relatively small, stable, genetically homogeneous population. Also, the overall similarity between RFLP profiles of the H. pylori isolates from Texas and Minas Gerais coincides with the fact that although geographically distanced, these populations are similar in being large, dynamic, and genetically heterogeneous. PMID- 10382121 TI - Seroprevalence of Helicobacter pylori and length of stay in a nursing home. AB - BACKGROUND: Helicobacter pylori infection appears to be contracted mainly in childhood, and it is associated with disadvantaged socioeconomic conditions, overcrowding, and living in institutions. In this study we determined the seroprevalence of H. pylori among elderly patients (age > or = 70 years) admitted to a major medical center in Israel, and studied the relationship between seroprevalence of H. pylori and the duration of stay in a nursing home prior to the admission. PATIENTS AND METHODS: Whole blood from 182 consecutive patients hospitalized at the Rabin Medical Center was tested for the presence of anti-H. pylori IgG using Helisal Rapid Blood Test kit (Cortecs Diagnostics). Multivariate logistic regression analysis was used to study the relation between H. pylori seropositivity and possible predictive factors such as age, gender and duration of stay in a nursing home. RESULTS: Of the 182 patients included in the study, 80 (44%) were living in nursing homes (NH) and 102 (56%) were living in their own homes (H) prior to admission. Subjects that stayed in nursing homes for more than 15 months were significantly more likely to be seropositive than subjects with a shorter duration of stay (84% and 63% respectively, p = 0.03). Using a multivariate logistic regression analysis on both the NH group and the whole group, seropositivity was found to be significantly associated with duration of stay in a nursing home (p = 0.03 and p = 0.01 respectively). Seropositivity was not associated with age in either group. CONCLUSIONS: Living in a nursing home is associated with increased risk for H. pylori infection in the elderly. There is a strong correlation between the duration of stay in a nursing home and the prevalence of H. pylori infection. PMID- 10382122 TI - Gastric ulcer formation after the Hanshin-Awaji earthquake: a case study of Helicobacter pylori infection and stress-induced gastric ulcers. AB - BACKGROUND: Both Helicobacter pylori (H. pylori) infection and various stresses are known to induce peptic ulcer disease of the upper gastrointestinal tract. However, the pathogenetic relationship between the two factors has not yet been clarified. We conducted a case-control study to examine whether H. pylori infection played a role in the development of gastric ulcer (GU) induced by life event stresses that were experienced after the Hanshin-Awaji earthquake. MATERIALS AND METHODS: Serum samples from patients in the devastated area who developed GUs during the 2 months following the Hanshin-Awaji earthquake and those from GU patients in the same area during the corresponding period of the previous year, and from gender-, age- and institute-matched ulcer-free controls were tested for the presence of the H. pylori IgG antibody. RESULTS: A significant association between H. pylori infection and the development of GU in uninjured patients was observed in all sets [matched odds ratio (OR) = 3.23, 95% confidence interval: 1.95-5.35]. Moreover, the prevalence of H. pylori infection in patients who developed GUs after the earthquake was not different from that for GU patients in the previous year. In contrast, there was no association between H. pylori infection and the development of GU in the physically injured patients after the earthquake. CONCLUSIONS: H. pylori infection may play an important role in the development of GUs that are induced by emotional life-event stresses. PMID- 10382123 TI - Prevalence of CagA, VacA antibodies in symptomatic and asymptomatic children with Helicobacter pylori infection. AB - BACKGROUND: Limited data are available on the prevalence of CagA and VacA Helicobacter pylori antibodies in children. The aim of this study was to investigate the antibody prevalence to the H. pylori virulence factors CagA and VacA in symptomatic and asymptomatic children with H. pylori infection and to correlate these antibodies with the severity of gastric inflammation or density of H. pylori organisms in the gastric mucosa. MATERIALS AND METHODS: Twenty-three symptomatic children and 132 asymptomatic children with positive H. pylori serology participated in this study. Anti-H. pylori IgG antibody and CagA or VacA H. pylori antibodies were measured by enzyme immunoassay (HM-CAP; sensitivity and specificity > 90%) and Western immunoblot (Helicoblot 2.0) methods, respectively. Gastric inflammation and H. pylori density were graded histologically using the revised Sydney criteria. RESULTS: The prevalence of CagA and VacA antibodies were 69% and 35% in symptomatic children and 54% and 52% in asymptomatic children, respectively. Multiple regression analysis showed a correlation between CagA antibody and the severity of gastritis but no correlation with other histological features, including the number of neutrophils or lymphoid follicles. Neither antibody correlated with the degree of bacterial density in the gastric mucosa. CONCLUSION: CagA and VacA H. pylori antibodies are common in the pediatric population. The combined CagA/VacA antibodies correlated weakly with the degree of mucosal inflammation. PMID- 10382124 TI - Pretreatment antibiotic resistance in Helicobacter pylori infection: results of three randomized controlled studies. AB - BACKGROUND: Although combinations of antibiotics and antisecretory drugs are useful for treatment of Helicobacter pylori infection, treatment failure is common. The aim of this study was to evaluate the relation between pretreatment antibiotic resistance and outcome by using six different treatment regimens for H. pylori infection. PATIENTS AND METHODS: Three hundred sixty-nine consecutive H. pylori-infected patients with dyspeptic symptoms were enrolled in three consecutive randomized, controlled, single-center clinical trials: trial A, 128 patients; trial B, 125 patients; trial C, 116 patients. Treatments consisted of (A) a 15-day course of dual therapy (omeprazole, 20 mg bid, and amoxicillin, 1 gm bid, or clarithromycin, 500 mg tid) (OA vs OC); (B) a 7-day triple therapy of omeprazole, 20 mg bid, plus metronidazole, 500 mg bid, and amoxicillin, 1,000 mg bid, or clarithromycin, 500 mg tid (OMA vs OMC); or (C) omeprazole, 20 mg bid, plus metronidazole, 500 mg bid, plus tetracycline, 500 mg qid, or doxycycline, 100 mg tid (OMT vs OMD). Diagnostic endoscopy was made in all patients before and 5 to 6 weeks after therapy. Six biopsies were taken from each patient for histology, rapid urease test, and H. pylori culture; antibiotic susceptibility testing was performed using the E-test method. RESULTS: Overall cure rates were poor for both dual therapies OA and OC (38% and 37%, respectively) and for triple therapies OMA, OMC, and OMD (57%, 55%, and 58%, respectively). The OMT combination was successful in 91% (95% confidence interval [CI], 80.4%-97%). Metronidazole resistance was present in 29.7% (95% CI, 24%-35%), amoxicillin resistance was present in 26% (95% CI, 21%-32%), clarithromycin resistance was present in 23.1% (95% CI, 18%-29%), tetracycline resistance was present in 14% (95% CI, 10%-20%), and doxycycline resistance was present in 33.3% (95% CI, 21% 47%). Antibiotic resistance markedly reduced the cure rates and accounted for most of the poor results with the triple therapies: 89% versus 23%; 77% versus 26%; 100% versus 60%; and 67% versus 23% for OMC, OMA, OMT, and OMD, respectively. OMT appeared to be the best because of the high success rate with metronidazole-resistant H. pylori (71%) and in low-level tetracycline resistance. CONCLUSIONS: Pretreatment antibiotic-resistant H. pylori can, in part, explain the low cure rate of the infection and the variability in outcome in reported trials. PMID- 10382125 TI - Combined activity of azithromycin and lansoprazole against Helicobacter pylori. AB - BACKGROUND: Due to its unique pharmacokinetic properties, azithromycin may be an attractive combination partner for H. pylori eradication regimens. However, up to 15% of clinical isolates are primarily resistant to azithromycin as well as to other macrolide antibiotics. Combination therapy with lansoprazole, a proton pump inhibitor known to have intrinsic antibacterial activity against H. pylori, may be useful to counteract such resistance. We therefore evaluated the combined effects of azithromycin and lansoprazole in vitro. MATERIALS AND METHODS: Minimal inhibitory concentrations (MICs) of azithromycin and lansoprazole alone and in combination were determined for 106 clinical H. pylori isolates by means of an agar dilution technique. Killing kinetics of seven isolates were also studied in fluid medium. RESULTS: MIC values for 50 and 90% of the isolates (MIC50, MIC90) were 0.19 and 0.5 mg/l for azithromycin, and 44.5 and 104 mg/l for lansoprazole. Nine strains (8.5%) had an MIC of azithromycin > or = 16 mg/l and were regarded as resistant. An additive interaction between the two drugs was found in 72 (68%), and indifferent effects in 24 strains (23%). Three of 9 azithromycin resistant strains regained sensitivity in the presence of lansoprazole. In fluid culture, synergism between the two drugs occurred in 6 out of 7 strains tested. CONCLUSION: In the majority of strains, lansoprazole and azithromycin interacted in an additive or synergistic manner depending on the test method employed. Addition of lansoprazole restored in vitro sensitivity to azithromycin in 3 out of 9 azithromycin-resistant strains. Such effects may enhance the elimination of H. pylori during clinical eradication therapy. PMID- 10382126 TI - Twice-daily versus thrice-daily clarithromycin in combination with ranitidine bismuth citrate in the eradication of Helicobacter pylori. AB - BACKGROUND: Ranitidine bismuth citrate (RBC), 400 mg bid for 4 weeks, plus clarithromycin, 500 mg tid, is a regimen approved by the US Food and Drug Administration for the eradication of Helicobacter pylori in patients with duodenal ulcers. Proof that the clarithromycin portion of the regimen could be given twice daily without loss of efficacy would reduce cost and improve patient compliance. The objective of this study was to compare the H. pylori eradication rates in patients who had duodenal ulcer and were randomly assigned to 4 weeks of treatment with RBC, 400 mg bid, in conjunction with 2 weeks of therapy with either clarithromycin, 500 mg tid, or clarithromycin, 500 mg bid. PATIENTS AND METHODS: Patients who had a duodenal ulcer and were H. pylori-positive by at least two tests were randomly assigned to (1) RBC, 400 mg bid for 4 weeks, plus clarithromycin, 500 mg tid for 2 weeks, or (2) RBC, 400 mg bid for 4 weeks, plus clarithromycin, 500 mg bid for 2 weeks. H. pylori eradication was assessed 4 weeks after completion of RBC plus clarithromycin. RESULTS: Three hundred eighty three patients from 78 centers had a duodenal ulcer and were H. pylori-positive. The modified intent-to-treat (MITT) and the per-protocol (PP) eradication rates were statistically equivalent between the twice-daily (65% MITT, 74% PP) and thrice-daily (63% MITT, 73% PP) clarithromycin treatment regimens. Incidence and types of adverse events did not differ between the two groups. CONCLUSIONS: For eradicating H. pylori in patients with duodenal ulcer, clarithromycin, 500 mg bid for 2 weeks, with RBC, 400 mg bid for 4 weeks, is equivalent to clarithromycin, 500 mg tid with RBC. The potential enhancement of patient compliance, reduced cost of clarithromycin, and equivalent efficacy would support the use of twice daily clarithromycin in triple-therapy regimens with RBC. PMID- 10382127 TI - Colloidal bismuth pectin: an alternative to bismuth subcitrate for the treatment of Helicobacter pylori--positive duodenal ulcer. AB - BACKGROUND: Bismuth triple therapy provides consistently good results in Helicobacter pylori eradication worldwide, whereas quadruple therapy using a combination of omeprazole and bismuth triple regimen has produced cure rates in excess of 90%. The prevalence of metronidazole-resistant strains was 26.8% in our area. Colloidal bismuth pectin (CBP) is a new, lower-priced bismuth salt made in China. The purpose of this study was to investigate the efficacy and safety of CBP triple and quadruple regimens in the treatment of H. pylori-positive duodenal ulcer. MATERIALS AND METHODS: In this prospective trial, 205 patients with H. pylori-positive duodenal ulcer were allocated randomly to receive one of four regimens: metronidazole, 200 mg; amoxicillin, 250 mg; and colloidal bismuth subcitrate (CBS), 120 mg (group 1), or CBP, 100 mg qid (group 2) for 2 weeks, then continued CBS, 240 mg, or CBP, 200 mg bid for a further 2 weeks. A quadruple regimen using a combination of omeprazole, 20 mg bid, and CBS triple therapy (group 3) or CBP triple therapy (group 4), respectively, was given to patients for 1 week, followed by omeprazole, 20 mg once daily for a further 3 weeks. Further endoscopy was performed at least 4 weeks after cessation of the treatment. H. pylori status was determined by histology, a 14C urea breath test, and a urease test. RESULTS: The per-protocol H. pylori cure rates were 85% (22 of 26 patients), 90% (35 of 39), 96% (46 of 48), and 95% (75 of 79) for groups 1 through 4. In the intention-to-treat analysis, cure rates were 79% (22 of 28), 83% (35 of 42), 90% (46 of 51), and 89% (75 of 84), respectively. The cure rates of quadruple therapy were higher than those of triple therapy; an 8.2% difference was not statistically significant (95% confidence interval [CI], 2.3-18.7%). The ulcer-healing rates were 88%, 87%, 98%, and 97%, respectively, for groups 1 through 4. The ulcer pain was relieved more rapidly in quadruple- than in triple therapy regimens. Two patients discontinued treatment prematurely owing to drug related side effects. CONCLUSION: One-week quadruple therapy is highly effective and safe in H. pylori eradication in Chinese patients. CBP is as effective as CBS. PMID- 10382128 TI - Randomized placebo-controlled trial of Helicobacter pylori eradication for iron deficiency anemia in preadolescent children and adolescents. AB - BACKGROUND: A few cases relating H. pylori infection to iron-deficiency anemia have been described recently. We investigated the role of H. pylori infection in iron-deficiency anemia in preadolescent children and adolescents. PATIENTS AND METHODS: We conducted a double-blind, placebo-controlled therapeutic trial in 43 subjects (mean age, 15.4 years) with iron-deficiency anemia. Endoscopy was performed, and biopsy specimens were examined by urease test and histological analysis. Twenty-two of 25 H. pylori-positive patients were assigned randomly to three groups. Group A patients were given oral ferrous sulfate and a 2-week course of bismuth subcitrate, amoxicillin, and metronidazole. Group B patients were given placebo for iron and a 2-week course of triple therapy. Group C patients were given oral ferrous sulfate and a 2-week course of placebo. Iron status was reassessed 4 weeks and 8 weeks after the 2-week regimen ended. RESULTS: Of the 43 subjects with iron-deficiency anemia, 25 (58.1%) had H. pylori in the antrum. Group A and B subjects, who received eradication therapy, showed a significant increase in hemoglobin level as compared with group C subjects at 8 weeks after therapy (p = .0086). CONCLUSIONS: Treatment of H. pylori infection was associated with more rapid response to oral iron therapy as compared with the use of iron therapy alone. Such treatment also led to enhanced iron absorption even in those subjects who did not receive oral iron therapy. PMID- 10382129 TI - Infection and myocardial infarction. PMID- 10382130 TI - Two intact executive capacities in children with autism: implications for the core executive dysfunctions in the disorder. AB - Many studies have shown that children with autism perform at a much lower level than control subjects on tests of executive functioning, defined as tasks requiring subjects to hold information in mind while suppressing a prepotent response. These tasks have invariably required subjects to (a) follow arbitrary and novel rules and (b) make a nonverbal response. We report that when one of these features is absent, children with autism are not impaired relative to controls. They perform at a similar level to normally developing children on the "tubes" task (containing no arbitrary and novel rules) and on the day/night task (in which the output is verbal). Results are consistent, at least, with the hypothesis that children with autism are challenged by executive tasks because they are unlikely to encode rules in a verbal form. PMID- 10382131 TI - Microcephaly and macrocephaly in autism. AB - Data from a series of 126 autistic children ages 2-16 years and referred to an Autism Diagnosis Unit in South-West France were examined. Macrocephaly (head circumference > 97th centile) was observed in 16.7% of the sample, a significantly higher proportion than that expected. Macrocephaly was more frequent among older subjects but was otherwise not associated with gender, developmental level, the presence of epilepsy or of medical disorders, or severity of autistic symptomatology. Microcephaly (head circumference < 3rd centile) was also significantly raised and found in 15.1% of the sample. Microcephaly was significantly associated with the presence of medical disorders. Results support those from recent studies suggesting a raised rate of macrocephaly in autism which, pooling published data, can be estimated to be 20%. It is argued that the raised incidence of microcephaly among low-functioning autistic subjects with medical disorders might have contributed to delay the recognition of an increased head circumference among a minority of subjects with idiopathic autism. PMID- 10382132 TI - Sexual behaviors in autism: problems of definition and management. AB - Surveys of sexual behavior in autism suggest a variety of behavioral expression. However, the course of sexual development in autism is unplotted, leaving questions about the normalcy of specific behaviors. Even less is known about deviations of sexual development and the incidence of paraphilias in this population. We explore the problems of definition of sexual behaviors and describe a case report that highlights the difficulties of management. An application of a testosterone-suppressing medication and its effect on sexual behavior are reported. After failure of behavioral and educational programs, leuprolide, an injectable antiandrogen, resulted in suppression of behaviors and retention of the participants' community placement. Follow-up for almost 3 years shows no abnormal physical effects. Dosage has been tapered over that period to a low but effective dose. Directions for research are discussed. PMID- 10382133 TI - A screening questionnaire for Asperger syndrome and other high-functioning autism spectrum disorders in school age children. AB - The high-functioning Autism Spectrum Screening Questionnaire (ASSQ) is a 27-item checklist for completion by lay informants when assessing symptoms characteristic of Asperger syndrome and other high-functioning autism spectrum disorders in children and adolescents with normal intelligence or mild mental retardation. Data for parent and teacher ratings in a clinical sample are presented along with various measures of reliability and validity. Optimal cutoff scores were estimated, using Receiver Operating Characteristic analysis. Findings indicate that the ASSQ is a useful brief screening device for the identification of autism spectrum disorders in clinical settings. PMID- 10382134 TI - Sleep patterns in autistic children. AB - Sleep disturbances are regarded as a common clinical feature in autistic children. This concept is based primarily on informal observations or studies conducted with questionnaires. In this study we compared data obtained by questionnaires to that obtained with actigraphy. Among 22 autistic children, 12 were reported as having sleep problems and 8 patients completed 72 hours actigraphy. While the employment of questionnaires disclosed that autistic children had an earlier morning awakening time and multiple and early night arousals, actigraphic monitoring showed that with the exception of an earlier morning arousal time (p = .045), sleep patterns of autistic children were similar to that of normal children. Parental oversensitivity to sleep disturbances of the autistic children may explain this phenomenon. PMID- 10382135 TI - Behavior problems of children with Down syndrome and life events. AB - Behavior problems of 44 children with Down syndrome between the ages of 6 and 15 and 44 controls without mental retardation matched for age, sex, and socioeconomic status were compared on the basis of mother and teacher ratings. Ratings from both sources indicated that children with Down syndrome had more behavior problems, in particular attention deficit, noncompliance, thought disorder, and social withdrawal. Life events from the past year were significantly associated with mother but not teacher ratings of Down syndrome behavior problems. PMID- 10382136 TI - Brief report: a pilot open clinical trial of intravenous immunoglobulin in childhood autism. PMID- 10382137 TI - Brief report: incidence of and risk factors for autistic disorder in neonatal intensive care unit survivors. AB - We investigated prospectively the incidence of autistic disorder (AD) in the neonatal intensive care unit and the risk factors associated with autistic development. The study population included the 5,271 children at St. Mary's Hospital and the diagnosis of AD was performed using DSM-III-R criteria. A total of 36 prenatal, perinatal, and postnatal factors were evaluated in the patients with AD, 57 cerebral palsy (CP), and 214 controls. AD was identified in 18 of the 5,271 children and the incidence was 34 per 10,000 (0.34%). This value was more than twice the highest prevalence value previously reported in Japan. Children with AD had a significantly higher history of the meconium aspiration syndrome (p = .0010) than the controls. Autistic patients had different risk factors than CP. PMID- 10382138 TI - Brief report: six months continuation treatment in naltrexone-responsive children with autism: an open-label case-control design. PMID- 10382140 TI - Brief report: autism and Aarskog syndrome. PMID- 10382139 TI - Brief report: specific executive function profiles in three neurodevelopmental disorders. PMID- 10382141 TI - Cycles and epicycles of autism. PMID- 10382142 TI - Can you explain the difference between autism and Asperger syndrome. PMID- 10382144 TI - Some people are born with stage fright. PMID- 10382143 TI - Acoustic studies of dysarthric speech: methods, progress, and potential. AB - EDUCATIONAL OBJECTIVES: (1) The reader will be able to describe the major types of acoustic analysis available for the study of speech, (2) specify the components needed for a modern speech analysis laboratory, including equipment for recording and analysis, and (3) list possible measurements for various aspects of phonation, articulation and resonance, as they might be manifest in neurologically disordered speech. PMID- 10382145 TI - Yoga, meditation, help teen sex offenders. PMID- 10382146 TI - Fact v fiction--dementia and aging. PMID- 10382148 TI - Pay attention, someone's watching your brain. PMID- 10382147 TI - Alcoholics' children--living with a stacked biochemical deck. PMID- 10382149 TI - Hug a tree, relieve your stress. PMID- 10382150 TI - Debate brews over caffeine addiction. PMID- 10382151 TI - Beware the three-martini lunch. PMID- 10382152 TI - Nurse-led art expression in addiction treatment. AB - Creative expression can be a potent force in recovery programs, and psychosocial nurses trained in group processes can use art expression as part of addiction recovery and relapse prevention. Enabling clients to understand recovery and recognize what life offers without addiction is one of the best defenses against relapse during difficult times. PMID- 10382153 TI - Health status risk factors of people with severe and persistent mental illness. AB - High rates of medical comorbidity and premature death have become normative health outcomes for individuals with mental illness. On average, people with mental illness die 10 to 15 years earlier than the general population. To date, little research and programmatic attention has focused on the health promotion and prevention needs of people with mental illness. Many factors have been cited as contributing to this problem, including the stigma of being mentally ill, poverty, and limited knowledge and access to health promotion services. Future health planning interventions should restructure the funding mandates of current health care delivery system from an illness and treatment model to one of illness prevention and health promotion. PMID- 10382154 TI - The family educator: a professional resource for families. AB - The family educator works with the client's family members to help in their understanding of their ill relative and to assist with managing the stress of the caregiver role. Families come to understand that they can join forces with the clinical team in helping their relative engage in a trial of treatment. Each treatment recommendation is explained to the family and decisions are arrived at by consensus through partnership meetings. Role modeling effective interaction approaches and providing assistance in generating alternative solutions to problematic situations are important functions of the family educator. PMID- 10382155 TI - Violence against nurses in outpatient mental health settings. AB - Nurses in outpatient mental health settings who have been assaulted may have an increased sense of vulnerability. Assault and verbal threats influence how nurses view client behavior. Mechanisms need to be developed to protect staff in outpatient settings and to support colleagues when assaults or threats occur. PMID- 10382156 TI - Suicides in an Australian inpatient environment. AB - Associated stressful life events both within and outside the hospital environment may contribute to the high-risk environment. The cohort studied here was hospitalized for a significant psychiatric problem, a factor which puts them in the high-risk category. When the severe psychiatric diagnosis is combined with a significant stressful life event, the risk of suicide is increased significantly. PMID- 10382157 TI - Pitfalls of the golden rule in caregiving. PMID- 10382158 TI - Whole-body MR imaging. Practical issues, clinical applications, and future directions. AB - Whole-body MR imaging is in evolution, and although accepting and recognizing limitations, it is likely that both technique and incurred acquisition times will shorten over the next decade. Although the development of dedicated whole-body MR scanners appears to offer the greatest promise for the future, the development of moving table tops, optimized pulse sequences, and advances in gradient technology now facilitate practical whole-body MR imaging using existing clinical systems. PMID- 10382159 TI - SMASH imaging. AB - SMASH imaging is a new MR imaging technique that can be used to multiply the speed of existing imaging sequences. It operates by using an array of radiofrequency (RF) detection coils to perform some of the spatial encoding normally accomplished with magnetic field gradients. The speed of the SMASH technique results from appropriate combinations of coil array RF signals in which multiple lines of image data are gathered simultaneously, rather than one after another. SMASH can be used in conjunction with most rapid imaging sequences, including EPI, resulting in multiplicative gains in imaging speed. This article reviews the basic principles of SMASH imaging, outlines requirements for practical implementation, and presents a variety of in vivo results, highlighting ways in which SMASH may be used to increase imaging speed and to improve image quality for clinical MR imaging applications. PMID- 10382160 TI - New MR imaging contrast agents. AB - Many new MR contrast agents are undergoing laboratory development, are in clinical trials, or are being used for routine clinical applications. These agents function by targeting specific receptors that allow the agent to accumulate within the reticuloendothelial system, the hepatobiliary system, or the blood pool. Each agent has unique properties that offer advantages over unenhanced and gadolinium chelate-enhanced MR imaging. Use of these agents, however, requires an understanding of their current and potential clinical indications and inherent limitations. This article reviews the development, evaluation, and clinical application of a number of these agents, including ferumoxides, Mn-DPDP, AMI-227, SHU-555A, Gd-EOB/DTPA, Gd-BOPTA, and MS-325. PMID- 10382161 TI - Clinical applications of half-Fourier (HASTE) MR sequences in abdominal imaging. AB - Endoscopic retrograde cholangiography remains a valuable technique in biliary disease, because therapeutic intervention, such as stone extraction and biliary drainage, can be carried out at the same time as diagnosis. Spatial resolution is superior to that of noninvasive imaging methods. HASTE MR cholangiopancreatography is as sensitive as sonography in detecting cholelithiasis. It is superior to sonography in diagnosing common bile duct stones, malignant biliary obstruction, and benign pancreatic disease. The noninvasive nature of HASTE MR imaging insures an expanding role in imaging patients with suspected pancreaticobiliary disease. PMID- 10382162 TI - Motion artifact control in body MR imaging. AB - The mechanisms involved in the generation of motion artifacts in MR imaging are complex and depend both on the type and direction of motion as well as on the parameters of the imaging sequence chosen. The methods used to control or reduce motion artifacts are multiple and the appropriate method for use with any given clinical situation will depend on the particular hardware and software of the MR imaging unit, the patient's clinical status, and the specific organ or disease state to be imaged. Some general guidelines for clinical use that are applicable in most scenarios can be defined, although preferences for the different techniques vary. Appropriate T1-weighted images of the upper abdomen and liver can be obtained with breath-hold T1-weighted gradient echo. These images should be acquired with inferior-superior spatial presaturation pulses to reduce vascular pulsation artifact and ghosting. The application of GMN will depend on the individual MR imaging system. If sufficient coverage cannot be obtained with gradient-echo imaging, then conventional T1-weighted images with phase-encoding reordering is suggested. The addition of spatial presaturation pulses (inferior superior) may be valuable. The use of fat suppression will further improve image quality by reducing ghost artifact and improving CNR, although SNR will decrease. T2-weighted imaging of the upper abdomen will depend greatly on the hardware and software of the MR imaging unit. Recent techniques of breath-hold T2-weighted imaging require faster and stronger gradients, and may not be universally available. If available, these techniques provide excellent anatomic detail, although image contrast (e.g., liver to spleen) may decrease. Respiratory triggered FSE techniques are the preferred method of imaging in most centers, because the imaging time is considerably less than conventional T2-weighted imaging whereas the image quality is improved. Liver lesion detection capability of the various techniques is still under study. The addition of fat suppression appears to improve image quality further with an increase in lesion detection. By understanding the principles underlying motion artifacts, one can choose the appropriate method of artifact control tailored for the individual clinical situation. In addition, the recognition of the variable appearances of motion artifacts will prevent interpretive errors and misdiagnoses. Careful attention to motion artifact reduction techniques can greatly improve patient care. PMID- 10382163 TI - Black-blood MR angiography. Techniques, and clinical applications. AB - As there are limitations in WB-MR angiography, so there are limitations in BB-MR angiography. Vessel morphology is visualized by means of the innermost nonattenuated layer of tissue, which, under ideal conditions, coincides with the luminal surface of the vessel wall. Vessel morphology may be depicted inaccurately whenever a portion of the vessel wall is undetectable with the MR imaging technique used. In such cases, vessel segments with exaggerated lumen diameter may result at locations where tissues with either a very short T2 or a low proton density are present. Another phenomenon that could potentially degrade the accuracy of vessel depiction with BB techniques is the effect of slowly flowing blood near the vessel walls. Residual blood signal would result in apparent vessel narrowing. Preliminary clinical experience in the brain, however, suggests that this adverse effect is less prominent with a turbo-SE-based BB technique than with a TOF WB technique. BB-MR angiography data sets may also present image postprocessing difficulties arising from the isointensity between the vessels and other dark structures such as bones and air-filled cavities. A limitation that is more specific to hybrid-SE-based BB-MR pulse sequences, particularly for very high spatial resolution applications, stems from the comparatively high RF specific absorption rates that result from the intensive use of 180 degrees refocusing pulses. GRASE-based BB-MR techniques that generate a fraction of the RF energy constitute a promising alternative for very high spatial resolution applications. In summary, to be effective, a BB technique must produce strong signal attenuation from flowing spins, ideally to the level of the baseline noise. Simultaneously it should produce good depiction of tissues with the comparatively short T2s characteristic of vessel walls and muscle, hence the need to operate with the shortest possible TE. Finally, high spatial resolution combined with fast data acquisition are requisites for imaging small vessels in the presence of motion, such as the carotid arteries. The flow properties of BB MR angiographic sequences that meet these criteria were reviewed for different anatomic locations. PMID- 10382164 TI - Advances in abdominal, aortic, and peripheral contrast-enhanced MR angiography. AB - This article reviews the basic principles of contrast-enhanced MR angiography, including methods used for sequence optimization and bolus timing, and describes clinical applications of contrast-enhanced MR angiography in the aortic, abdominal, and peripheral arteries. Novel MR angiography imaging techniques also are described, including moving table-top MR angiography, MR fluoroscopy, and time-resolved MR angiography. PMID- 10382165 TI - MR-guided therapy. Applications in the abdomen. AB - This article provides an overview of current abdominal interventional applications that use MR imaging guidance and monitoring for diagnostic biopsies, tumor ablations and--owing to the development of new pulse sequences--monitoring the ablation process, and aspiration and drainage of fluid collections. These applications are optimal for patients with lesions that can be localized only by MR imaging or for lesions in suboptimal locations such as the dome of the liver. PMID- 10382166 TI - Virtual endoscopy in abdominal MR imaging. AB - As illustrated in this review, the diagnostic utility of VIE is highly dependent on the individual application. Virtual intraluminal endoscopy does not alter the informational content of the data set; rather, the data is presented in a different manner. In view of the considerable time requirements still associated with the VIE rendering process, the additional diagnostic value gained by VIE must be determined for each of the potential applications. Thus VIE was shown to be of no additional value in the assessment of 3D MR angiographic data sets of the abdominal aorta and renal arteries. For MR colonography, on the other hand, in vitro experiments as well as in vivo experience have demonstrated a vast diagnostic value as a primary screen supplemented by detailed analysis of reformatted images in all three orthogonal axes. In summary, virtual intraluminal imaging is a powerful tool for exploring 3D MR data sets of various abdominal structures from a different perspective. Clearly much more work is required to determine its diagnostic utility relative to its cost. PMID- 10382167 TI - Coronary MR angiography. AB - Coronary MR angiography has developed rapidly over the past several years. Not only is research being performed at academic centers but industry is also investing in dedicated contrast agents and cardiac MR imaging platforms. Although current coronary MR angiography has limited clinical utility, its place within the assessment of ischemic cardiac disease is evolving. The technology currently under investigation holds much promise, especially when one considers that MR has the potential to provide information currently supplied by the performance of a number of screening tests. It would be far more cost-effective to perform a single MR examination than to perform a stress echo, rest-stress nuclear medicine examination and a conventional coronary angiogram. In addition, clinicians need information about coronary flow and myocardial perfusion. Although some of this information can be currently obtained with an intravascular Doppler flow wire or by positron emission tomography, MR angiography offers the advantages of being both noninvasive and more easily accessible in comparison to either method. The combined information promised by a comprehensive cardiac MR examination that includes coronary MR angiography as a component is an exciting prospect. PMID- 10382168 TI - Ventilation-perfusion MR imaging of the lung. AB - The assessment of regional ventilation in human lungs is important for the diagnosis and evaluation of a variety of pulmonary disorders, including pulmonary emphysema, diffuse lung disease (e.g., sarcoidosis, and pulmonary fibrosis), lung cancer, and pulmonary embolism. This article introduces new MR imaging techniques of pulmonary ventilation and perfusion that will provide a framework for assessing regional pulmonary functions of the lung. PMID- 10382169 TI - Pulmonary MR angiography. AB - Imaging of the pulmonary vasculature with "inflow" MR imaging is difficult both to perform and interpret. Over the last few years, however, a greater understanding of contrast-enhanced techniques and the availability of fast gradient performance have facilitated the development of high resolution breath hold images with high contrast-to-noise ratios. Increasing clinical experience with these contrast-enhanced techniques suggests a likely role for MR angiography in investigating patients with a variety of pulmonary vasculature disorders, including pulmonary embolism, pulmonary hypertension and arterio-venous malformation. PMID- 10382170 TI - Vascularity assessment of breast lesions with gadolinium-enhanced MR imaging. AB - Gadolinium-enhanced MR imaging techniques can be used to assess both breast tumor morphology and vascularity. Pharmacokinetic models can be used to extract physiological parameters related to tumor vascularity from signal intensity-time curves. This article describes an empirical method, using three-point high resolution MR imaging, that also provides assessment of tumor vascularity. These techniques appear to improve diagnostic specificity of breast MR imaging and provide a noninvasive method for tumor characterization that may have prognostic value. PMID- 10382171 TI - Cancer as metaphor. PMID- 10382172 TI - Stovall receives Achievement Award for Outstanding Contributions to Cancer Care. PMID- 10382173 TI - Raymon receives NurseWeek Award for Innovation. PMID- 10382174 TI - The role of octreotide in malignant bowel obstruction. PMID- 10382175 TI - The role of the advanced practice nurse in a genitourinary/oncology program. PMID- 10382176 TI - Development of a nurse-managed colon cancer screening center. PMID- 10382177 TI - Anticipating blood transfusions in the home: a safe and efficient approach. PMID- 10382178 TI - Simple relaxation techniques for patients with cancer and staff. PMID- 10382179 TI - Ovarian cancer early detection program. PMID- 10382180 TI - Testing forms with transplant protocols. PMID- 10382181 TI - The value of interdisciplinary care. PMID- 10382182 TI - Patient education: reaching the Hispanic community. PMID- 10382183 TI - A national survey of certified, recertified, and noncertified oncology nurses: comparisons and contrasts. AB - PURPOSE/OBJECTIVES: To explore opinions about the OCN credential, the ways in which it was obtained and retained, and the extent to which it is valued by employers. DESIGN: A descriptive comparison study using a cross-sectional survey design. SAMPLE: Questionnaires were mailed to a nationwide sample of 2,429 RN members of the Oncology Nursing Society; 1,217 (50%) surveys were returned. The majority of respondents were female, 30-49 years of age. Caucasian, and had practiced nursing for more than 11 years. MAIN RESEARCH VARIABLES: Certification status, work role characteristics, preparation strategies for the certification examination, and motivation for obtaining certification. FINDINGS: Oncology nurses recognize the importance and value of OCN certification. The primary reasons oncology nurses obtain and retain certification include the desire for personal achievement, professional growth, and development. OCNs were more likely to work in a setting where the employer supports professional development through continuing nursing education. IMPLICATIONS FOR NURSING PRACTICE: Because health care is increasingly delivered in ambulatory/home settings and the population is aging, oncology certification needs to be encouraged among nurses who work in these settings or with geriatric populations. Certified nurses tended to experience more job satisfaction than noncertified nurses. PMID- 10382184 TI - Web site design and development issues: the Washington State Breast and Cervical Health Program Web Site Demonstration Project. AB - PURPOSE/OBJECTIVES: To explore the development of a customized Web site to assist Breast and Cervical Health Program (BCHP) outreach staff in a community screening program and to evaluate the Internet knowledge and access issues and barriers of outreach staff during a two-year period using the Web site. DESIGN: Knowledge, access issues and barriers, and descriptive questionnaires. SETTINGS: Comprehensive cancer center in Seattle, WA, workshops, and presentations around the state. SAMPLE: BCHP outreach workers, screening coordinators, and almost exclusively public health nurses from regional health districts and program contracted clinics. METHOD: Web site development was based on continuous input from sample. Detailed descriptions of computer and Internet resources and opinions about the use and usefulness of the BCHP Web site came from a 1996 evaluation and 1998 follow-up conducted using mailed and online Web questionnaires. "Hits" to the Web site were monitored monthly. MAIN RESEARCH VARIABLES: Computer and Internet resources were used along with monthly Web site traffic and opinions about the use and usefulness of the BCHP Web site in the outreach program. FINDINGS: Use of the BCHP Web site has risen steadily over two years to reach a stable plateau. User evaluations show a marked increase in the adoption of the Internet as a working tool. Users believe the Internet is becoming increasingly important to their work. More training and familiarization with the Web is needed. CONCLUSIONS: The Web is an efficient medium for improving communication and providing easy access to resources within the BCHP program. IMPLICATIONS FOR NURSING PRACTICE: Public health programs with meager resources can benefit from the relatively inexpensive use of customized and versatile Web sites. PMID- 10382185 TI - Analysis of end-of-life content in nursing textbooks. AB - PURPOSE/OBJECTIVES: To determine the amount and types of content regarding pain and end-of-life (EOL) care included in major textbooks used in nursing education. DESIGN: Descriptive. SAMPLE: 50 texts (45,683 pages) selected from a potential of more than 700 texts. METHODS: Content analysis and quantification of nine essential areas of EOL care content present in the texts. MAIN RESEARCH VARIABLES: Nine areas of EOL care: palliative care defined; quality of life, pain; other symptom assessment/management; communication with dying patients and families; role/needs of family caregivers in EOL care; death; issues of policy, ethics, and law; and bereavement. FINDINGS: Only 2% of the overall content and 1.4% of chapters in nursing texts were related to EOL care. Based on the analysis, many deficiencies were identified in the texts, including inaccurate information and a lack of information regarding critical EOL topics. CONCLUSIONS: Nursing texts contain limited content regarding EOL care. Increased attention to this area is essential in preparing nurses to care for patients at EOL. IMPLICATIONS FOR NURSING PRACTICE: Nursing practice is based on the foundation of nursing education. Changes in nursing school curriculum and provision of continuing education for practicing nurses are essential for improved EOL care. PMID- 10382186 TI - Cervical cancer screening knowledge, behaviors, and beliefs of Vietnamese women. AB - PURPOSE/OBJECTIVES: To describe the knowledge, beliefs, and practices of cervical cancer screening of Vietnamese women who have migrated to the United States. DESIGN: Exploratory, descriptive. SETTING: Five Vietnamese churches in southeastern Louisiana. SAMPLE: Nonprobability sample of 96 adult Vietnamese migrant women. METHODS: Data were collected by a bilingual nurse during face-to face interviews conducted in the language preferred by the subjects (English or Vietnamese). MAIN RESEARCH VARIABLES: Cervical cancer screening knowledge, behaviors, and beliefs. FINDINGS: Three fourths of the Vietnamese women interviewed could not correctly explain what a Pap test is used for, and few were aware that the most commonly occurring cancer in Vietnamese females in the United States is cervical cancer. Most believed that their risk of cervical cancer was low. Less than half reported ever having had a Pap test and cited not having a gynecologist, cost, and fear of the test as reasons for not ever having had the test done. CONCLUSIONS: The number of Vietnamese women who adhere to cervical cancer screening guidelines is low. Cultural beliefs and structural barriers influence the choices that Vietnamese women make regarding Pap test utilization. IMPLICATIONS FOR NURSING PRACTICE: When planning programs to promote cervical cancer screening, nurses must target those at greatest risk--Vietnamese women. Culturally sensitive educational interventions and cervical cancer screening programs for Vietnamese women are needed to increase Pap test utilization and the early detection of cervical cancer. PMID- 10382188 TI - The effect of nutritional supplements on food intake in patients undergoing radiotherapy. AB - PURPOSE/OBJECTIVES: To describe the effect of nutritional supplements on food intake in patients undergoing radiotherapy. DESIGN: Experimental prospective. SAMPLE: 40 newly diagnosed patients with cancer beginning external beam radiation therapy. METHODS: Weekly dietary counseling and recording of total daily dietary intake for three days a week for four weeks. One half of the subjects were randomly assigned to ingest a liquid nutritional supplement between meals and at bedtime. MAIN RESEARCH VARIABLES: Total daily protein and caloric intake, food derived protein and caloric intake, and supplement-derived protein and caloric intake. FINDINGS: Subjects ingesting nutritional supplements between meals significantly increased their total caloric and protein intake above that of controls and did not reduce their food-derived caloric or protein intake compared to controls. CONCLUSIONS: Nutritional supplements can be used to increase total caloric and protein intake without causing a significant reduction in food intake. IMPLICATIONS FOR NURSING PRACTICE: In this patient sample, supplements were not substituted for food intake. Further research is needed to determine the effects of supplements on appetite in patients with advanced cancer. PMID- 10382187 TI - The Index of Nausea, Vomiting, and Retching: a new format of the lndex of Nausea and Vomiting. AB - PURPOSE/OBJECTIVES: To determine the reliability of the Index of Nausea, Vomiting, and Retching (INVR), a new format of the Rhodes Index of Nausea and Vomiting Form 2 (INV-2). DESIGN AND SETTING: A parallel form study was conducted at a large, Midwestern teaching hospital and a cancer center. SAMPLE: Convenience sample of 159 subjects: 40 obstetrical, 60 oncological, 59 medical/surgical. METHODS: Two instruments, the INVR and the INV-2, were administered approximately 30-60 minutes apart. One-half of the subjects completed the INVR first, and the other half completed the INV-2 first. MAIN OUTCOME MEASURES: Equivalency measures of reliability correlation coefficients for both instruments. FINDINGS: A high rate of agreement was found in the responses between the two forms. In cases of clear disagreement, the responses to the INVR were more frequently consistent than the responses to the original form. CONCLUSIONS: INVR has tested reliability and is more user friendly for the patient and the healthcare provider. IMPLICATIONS FOR NURSING PRACTICE: Nurses have a focal role in managing symptoms. Managing nausea, vomiting, and retching requires excellent assessment skills of the patient's personal symptom experience and knowledge of pharmacology. Efficient, cost-saving assessments require accurate self-report instruments that permit patients to quantify their symptom experiences. The INVR can provide a scientific base from which to prescribe and teach patients and may improve their quality of life. Reliable and valid self-reporting instruments are essential for managing these adverse symptoms. PMID- 10382189 TI - Caring for patients who experience chemotherapy-induced side effects: the meaning for oncology nurses. AB - PURPOSE/OBJECTIVES: To explore the meaning for oncology nurses of caring for patients with cancer who experience chemotherapy-induced side effects. DESIGN: Descriptive phenomenology. SETTING: Participant's work setting (n = 2), home setting (n = 1), and social setting (n = 2). SAMPLE: Five female oncology nurses (four Caucasian and one African American), age range 31-62 years, with an average of 13.3 years in oncology nursing from a variety of settings (e.g., private medical practice, ambulatory infusion centers, university-based bone marrow/peripheral blood stem cell transplantation unit) with present or recent experience administering chemotherapy. METHODS: Open-ended, audiotaped interviews were conducted. The text was transcribed verbatim and was analyzed using Colaizzi's phenomenologic analysis technique. MAIN RESEARCH VARIABLE: Meaning for oncology nurses of providing care to patients with cancer experiencing chemotherapy-induced side effects. FINDINGS: Six main themes and four subthemes emerged from the data analysis. The main themes were Being Drawn Into Patients' Experiences of Suffering. Being Challenged by Personal and Professional Imperatives to Act, Defining Treatment Purpose for Self and Patient, Reconsidering the Meaning of "Sick" and "Well," Being Changed by Ties of Shared Experience, and Advocacy for Self and Patient. CONCLUSIONS: The fundamental meaning to oncology nurses of providing care to patients with cancer experiencing chemotherapy-induced side effects is their empathetic use of self as an oncology nurse/friend to alleviate the suffering related to these side effects. IMPLICATIONS FOR NURSING PRACTICE: Providing nursing care to patients who experience chemotherapy-induced side effects is both rewarding and stressful. Sharing these research results may help other oncology nurses discover and experience deeper meaning in their own practice. PMID- 10382190 TI - Side-effects burden, psychological adjustment, and life quality in women with breast cancer: pattern of association over time. AB - PURPOSE/OBJECTIVES: To describe the side-effects burden experienced over time by 53 women who were receiving treatment for breast cancer and to describe the association of side-effects burden with psychological adjustment and life quality. DESIGN: Data were drawn from the Self-Help Intervention Project (SHIP), an intervention study designed to test the effectiveness of nursing interventions for women receiving treatment for breast cancer. SETTING: Subjects were interviewed in their homes or treatment locations three times over a period of four to five months. SAMPLE: 53 women randomly assigned to the control group of the SHIP. METHODS: The researchers collected data after treatment was initiated, six to eight weeks later, and three months after that. MAIN RESEARCH VARIABLES: Side-effects burden, psychological adjustment, and life quality. FINDINGS: Fatigue was the most problematic side effect over time. Other problematic side effects included sore arm(s), difficulty sleeping, hair loss, and skin irritation. Significant associations were evident for psychological adjustment with symptom extension and number of side effects at Time 2 and Time 3. Depression burden and anxiety burden were associated significantly with psychological adjustment at all three times. Overall life quality and present life quality was associated negatively with symptom extension and number of side effects at all three times. Fatigue burden was associated negatively with life quality at Time 2 and Time 3 with depression burden and anxiety burden negatively associated with life quality at all three times. CONCLUSIONS: Over time, evidence showed that negative feelings, in particular depression burden and anxiety burden, persist. Depression burden and anxiety burden each were negatively associated with overall and present life quality at all three times. IMPLICATIONS FOR NURSING PRACTICE: A need exists for clinically individualized nursing interventions that will reduce the side effects burden of women receiving treatment for breast cancer. Interventions can do much to reduce the perception of illness severity so that psychological adjustment and life quality can be maintained. PMID- 10382192 TI - A trip through the ear in search of deafness. PMID- 10382191 TI - Relationship between fatigue and quality of life in patients with glioblastoma multiformae. AB - PURPOSE/OBJECTIVES: To evaluate changes in fatigue at the time of diagnosis (Time 1) and two weeks after the completion of radiation therapy (Time 2) and to determine the relationship between fatigue and quality of life (QOL) at each time interval in patients with glioblastoma multiformae (GBM). DESIGN: Descriptive study to evaluate fatigue and QOL. SETTING: Neuro-oncology clinic for accrual. Clinic, home, or office for patients interviews. SAMPLE: 60 adult patients diagnosed with GBM who received radiation therapy. METHODS: Participants completed a demographic data sheet, the Profile of Mood States, and the Multidimensional Quality of Life Scale-Cancer Version 2 at both time points. MAIN RESEARCH VARIABLES: Fatigue and QOL. FINDINGS: Fatigue significantly increased from Time 1 to Time 2 (t = -2.69, p = 0.009). Increases in fatigue were associated with significant decreases in QOL at Time 1 (r = -0.57) and Time 2 (r = -0.60). CONCLUSIONS: Patients with GBM experience increases in fatigue after radiation therapy. Increases in fatigue are associated with decreases in almost all aspects of patients' QOL. IMPLICATIONS FOR NURSING PRACTICE: Nurses must provide patients with information about the occurrence of fatigue during radiation therapy and recommend interventions to deal with this devastating symptom. PMID- 10382193 TI - Detection of hearing loss in children. PMID- 10382194 TI - Cochlear implants in children with sensorineural inner ear hearing loss. PMID- 10382195 TI - Diagnosis and management of otalgia in the pediatric patient. PMID- 10382196 TI - Detection and management of childhood cholesteatoma. AB - Childhood cholesteatoma is an aggressive disease that demonstrates higher rates of recidivism than its adult counterpart. The priorities in ideal management include total removal, followed by hearing restoration, followed by preserving the ear anatomy. Especially in children, one must endeavor to preserve ear anatomy if it does not jeopardize total removal of cholesteatoma. Absolute indications for CWD surgery include an only-hearing ear, a severely destroyed posterior canal wall, an extremely contracted mastoid, and matrix overlying a semicircular canal fistula. Reasons for staging childhood cholesteatoma include suspected residual disease, uncertainty about total removal of cholesteatoma, severe mucosal disease, and CWU procedures in which the cholesteatoma has diffusely invaded the bone. Adequate long-term follow-up is imperative. Patients and their families should be reminded frequently of the importance of close follow-up because recidivism is frequent. Successful management of cholesteatoma in children does not involve a rigid, "one-way" approach. The surgeon must be flexible and capable of employing the most appropriate procedure for the patient. PMID- 10382197 TI - The pediatrician's role in caring for patients with congenital microtia and atresia. PMID- 10382198 TI - Nonsurgical treatment of auricular deformities in neonates and infants. PMID- 10382199 TI - Management of traumatic auricular injuries in children. AB - Management of traumatic auricular injuries in children does not differ qualitatively from that in adults. The more common, simple injuries in children can be as readily treated by their pediatricians as those in adults are by their primary care physicians. However, children are more likely to need general anesthesia for significant repairs. A comprehensive initial assessment of the injury site remains the foundation for treatment planning. An appreciation for the variety of treatment options available for the injured ear should facilitate an interaction between the pediatrician and the surgeon when injury is complex and should lead to the best outcome for each child. PMID- 10382200 TI - Resident's column. The ear and hearing. PMID- 10382201 TI - Non-invasive diagnosis of acute heart- or lung-transplant rejection using radiolabeled annexin V. AB - BACKGROUND: Apoptosis is a ubiquitous set of cellular processes by which superfluous or unwanted cells are eliminated in the body without harming adjacent healthy tissues. When apoptosis is inappropriate (too little or too much), a variety of human diseases can occur, including acute heart or lung transplant rejection. OBJECTIVE: Our group has developed a new radiopharmaceutical, radiolabeled annexin V, which can image apoptosis. RESULTS AND CONCLUSION: Here we briefly review the biomolecular basis of apoptosis and its role in acute rejection. We also describe the possible use of radiolabeled annexin V to screen children noninvasively for acute rejection following organ transplantation. PMID- 10382202 TI - Hepatic arterial pseudoaneurysm: a rare complication of blunt abdominal trauma in children. AB - We report a child who developed a hepatic artery pseudoaneurysm following blunt hepatic injury. This is a rare complication of hepatic trauma in children. The imaging evaluation and clinical management of hepatic artery pseudoaneurysms are presented. PMID- 10382203 TI - A spectrum of segmental multicystic renal dysplasia. AB - BACKGROUND: Multicystic renal dysplasia (MCDK) is a common anomaly well described in the literature, but less well described when involving only a portion of a kidney. OBJECTIVE: To present the imaging spectrum, natural history and associated anomalies of six kidneys with segmental MCDK. MATERIALS AND METHODS: Five children with segmental MCDK (one with bilateral segmental MCDK) referred to our hospital between 1989 and 1996 were reviewed. All had at least one ultrasound examination. Four had a voiding cystogram and three had renal scintigraphy. RESULTS: Four children had antenatal diagnosis of cystic renal abnormality. In two, with obvious duplex kidneys and associated ureteroceles, the diagnosis of upper moiety MCDK was obvious either antenatally or immediately postnatally. In the other three there were diagnostic difficulties. One patient had bilateral widespread cysts obscuring the functioning renal portions. Another presented in utero with a large ureterocele and a cystic upper pole that had involuted by birth. The fifth had a nephrectomy at 3 years for a multiloculated cystic mass. Varying degrees of involution occurred in the five kidneys seen early. Reflux was demonstrated into the ipsilateral functioning lower moiety and midpole. CONCLUSION: In these children as in other studies, the commonest presentation of segmental MCDK is in the upper pole of a duplex kidney associated with a ureterocele at the end of the atretic ureter. Atypical segmental MCDK may present a diagnostic dilemma and should be included in the differential diagnosis of multiloculated cystic masses and cystic kidneys. PMID- 10382204 TI - Ultrasound findings in children with toxocariasis: report on 18 cases. AB - PURPOSE: To evaluate abdominal ultrasound (US) findings in children infected with Toxocara canis. MATERIALS AND METHODS: Eighteen children, 18 months to 7 years of age, with serological diagnosis of T.canis infection underwent abdominal US. Eosinophil counts, hemoglobin levels and immunoglobulin E titers were measured for all patients. RESULTS: Abdominal ultrasound revealed multiple hypoechoic areas in the livers of 15 patients (83.3%). Hepatohilar lymph-node enlargement was present in 14 patients (77.7%), 2 of whom also showed peripancreatic lymph node enlargement. Hepatomegaly was present in 13 patients (72.7%) and splenomegaly in 9 (50%). CONCLUSION: The most prevalent findings of abdominal ultrasound examination of children with T.canis infection are hepatic granulomas and abdominal lymph-node enlargement. This infection should be considered in the differential diagnosis of any children who exhibit these findings on abdominal US examination, especially for those with eosinophilia. PMID- 10382205 TI - Urachal neuroblastoma: first case report. AB - Tumours of the urachus are exceptional in children. They represent 0.01% of all tumours and consist of mucosecretory adenocarcinoma and, more rarely, transitional cell carcinoma. We report a 6-month-old child with a urachal mass which, following biopsy, was shown to be a neuroblastoma. PMID- 10382206 TI - Common phenotype and etiology in warfarin embryopathy and X-linked chondrodysplasia punctata (CDPX) PMID- 10382207 TI - The vascular vise causing TAPVR type I to radiographically mimic TAPVR type III. AB - A subtype of supracardiac total anomalous pulmonary venous return (TAPVR) consists of the vertical vein passing between the left pulmonary artery and the left mainstem bronchus resulting in relative obstruction to pulmonary venous return. This has been termed the vascular vise. In this situation, the supracardiac type of TAPVR (Type I) may mimic radiographically the infradiaphragmatic type (Type 3). PMID- 10382208 TI - Respiratory foreign bodies and Eikenella corrodens brain abscess in two children. AB - We report the coexistence of aspirated foreign bodies and brain abscess in two boys. One child had aspirated a metallic needle, and in the other boy partially embedded sunflower seeds were found in the bronchial wall. Both patients had growth of Eikenella corrodens (oral gram-negative flora) from the abscess. Aspirated foreign body in the respiratory tract should be one of the diagnostic considerations if any of the normal oropharyngeal organisms such as E. corrodens is the causative organism of brain abscess. PMID- 10382209 TI - Macromastia in a newborn with Alagille syndrome. AB - We present a case of macromastia in a newborn with Alagille syndrome. A review of the literature failed to find any prior reports of this findings in Alagille syndrome patients. We propose that this patient's macromastia may be related to her liver failure and abnormal estrogen metabolism. PMID- 10382210 TI - Growth disturbance of the sternum and pectus deformities: imaging studies and clinical correlation. AB - BACKGROUND: Radiologic reports of "normal" chest are not uncommon when there clearly are irregularities of sternal ossification and maturation. Analysis of imaging studies of sternal deformities for growth disturbances is not common in the literature and is addressed in this manuscript. OBJECTIVE: To determine the influence of sternal growth on development of pectus deformities and correlate imaging studies with clinical aspects of different types of these deformities. MATERIAL AND METHODS: One hundred forty-one children and adolescents with pectus deformities were evaluated. Sternal growth was estimated through the development of radiographic indices that were available for 57 patients with pectus deformities and for 71 controls. Magnetic resonance imaging of the sternum was performed in two patients to correlate with radiographic information. RESULTS: Radiographic indices of the sternum suggested growth disturbances in three basic types of pectus carinatum deformities: superior, inferior and lateral, and in the localized type of pectus excavatum. CONCLUSION: Sternal growth seems to have an important influence on the development of carinatum superior; partial influence on carinatum inferior, carinatum lateral, and excavatum localized; and no influence on excavatum wide pectus deformities. The endochondral growth of the sternum and costal arches is an important concept that aids in the interpretation of imaging studies and the orthopedic approach to management of these deformities in children and adolescents. PMID- 10382211 TI - CT-guided percutaneous needle biopsy of small lung nodules in children. AB - Malignant pulmonary nodules in a patient with neoplasia alters clinical staging and therapy. Although it may be difficult, it is imperative to obtain a tissue diagnosis of such nodules. We describe a coaxial technique using controlled respirations, which we have found useful for biopsy of small pulmonary nodules in children. Clinical records, imaging and pathology (histology and cytology) were reviewed for 17 children who had 18 nodules biopsied in the method described. A diagnosis of malignant or benign tissue was made in 15 nodules. One biopsy was inadequate and there was one false-negative. Adequate cores were obtained in 15 nodules, cytology in 2 and inadequate material in 1. There was no significant morbidity. PMID- 10382212 TI - Late neurosonographic screening is important to the diagnosis of periventricular leukomalacia and ventricular enlargement in preterm infants. AB - BACKGROUND: Recent cost-containment strategies suggest limiting screening neurosonograms to the second week of life in premature infants with lower gestational ages (< 30 weeks), birth weights (< 1250 g), or more complicated clinical courses. OBJECTIVE: To determine if such strategies reduce detection of cystic periventricular leukomalacia (cPVL) and persistent ventricular enlargement (pVE)--late sonographic abnormalities highly predictive of adverse neurodevelopment in preterm infants. METHODS: Timing, findings, and number of neurosonograms were reviewed for all survivors born at < or = 32 weeks' gestation at University Hospital, Denver, Colo., between January 1992 and June 1995. RESULTS: Of 236 surviving infants, 61 (26%) were never scanned, and 175 (74%) had a total of 432 scans. Only 106 infants (45%) had a neurosonogram on or after 28 days (timed to detect all cPVL/pVE). Eleven infants (4.7%) had cPVL, and 19 (8%) had pVE. Severity of clinical course did not predict development of cPVL, but was a better predictor of pVE. Initial neurosonograms were normal in 6/11 (55%) with cPVL and 5/19 (26%) with pVE. Screening declined from 86% of infants in 1992 (average 2.54 neurosonograms each), to 64% by 1994-1995 (average of 2.22 neurosonograms each). Infants > 30 weeks' gestation comprised 55 of 61 patients without any neurosonograms (90%), 4 of 11 patients with cPVL (36%), and 4 of 19 patients with pVE (21%). CONCLUSION: Screening neurosonography has declined from 1992 to 1995, particularly in larger premature infants (30-32 weeks' gestation) who remain at risk for cPVL and pVE. Clinical course or results of initial studies do not always predict the development of these late abnormalities. We recommend that one neurosonogram be done at > or = 4 weeks of age in all premature infants < or = 32 weeks' gestation, regardless of birth weight, clinical course, or results of prior studies. An earlier neurosonogram should be obtained for infants < 30 weeks' gestation in the second week of life to detect complications of intracranial hemorrhage. PMID- 10382213 TI - Cervical neurofibromas in children with NF-1. AB - BACKGROUND: Children with neurofibromatosis type 1 (NF1) are at increased risk of developing plexiform neurofibroma throughout the body, including the cervical soft tissues. However, the incidence of cervical soft tissue tumors and the value of screening MR for children with NF1 are not known. PURPOSE: The purposes of this study were to determine the incidence and clinical significance of cervical tumors seen on MR imaging in children with NF1. MATERIALS AND METHODS: A retrospective review of the brain and orbit MR with cervical images obtained on 95 children who meet the NIH consensus criteria for NF1 and who are followed at our neurofibromatosis clinic was carried out. RESULTS: Cervical tumors were found on MR imaging in 21 of 95 (22%) children. Of 21 children with cervical tumors, 14 children were determined to be surgical candidates. In nine children, MR imaging altered the clinical management by demonstrating tumors for which surgery was indicated, but the tumors were not suspected prior to MR imaging. CONCLUSION: Cervical tumors are commonly seen in children with NF1. MR imaging may demonstrate a significant number of tumors that require surgery, but were not suspected prior to MR imaging. PMID- 10382214 TI - Single central maxillary incisor with nasal pyriform aperture stenosis--CT diagnosis prior to tooth eruption. AB - Two children with nasal pyriform aperture stenosis had the diagnosis of single central maxillary incisor made with CT scanning prior to tooth eruption and the clinical appreciation of this finding. The surgical and clinical implications of this diagnosis will be presented. PMID- 10382215 TI - MRI evaluation of multifidus muscles in adolescent idiopathic scoliosis. AB - BACKGROUND: The role of the multifidus muscles in the initiation and progression of curve in adolescent idiopathic scoliosis is not fully understood and controversy exists as to the side of the abnormality. OBJECTIVE: To evaluate on MRI the multifidus muscles at the apex of the major curve in adolescent idiopathic scoliosis to ascertain if the multifidus muscles on the convex or concave side are abnormal and the relationship to curve severity. MATERIALS AND METHODS: Forty-six patients with adolescent idiopathic scoliosis, separated into two groups, were studied using a 1.5-T MR scanner with the synergy spine coil, employing a modified STIR (short tau inversion recovery) axial sequence obtained at the apex of the major scoliotic curve. RESULTS: No hyperintense signal change was demonstrated in the convex side multifidus muscles in any patient. In group I, 16 of 18 patients with severe or rapidly progressive curve showed increase in signal intensity in the multifidus muscle on the concave side of the apex of the curve. In group II, of the 15 patients with mild curve (Cobb angle 10-30 degrees), 4 had increased signal intensity in the multifidus muscle on the concave side; of the 13 with more severe curve (Cobb angle greater than 30 degrees), 10 had increase in multifidus signal intensity on the concave side. CONCLUSIONS: The concave-side multifidus muscle at the apex of a scoliotic curve was morphologically abnormal. A significant association between abnormal signal change and curve severity was also established. PMID- 10382216 TI - Juvenile rheumatoid arthritis and lymphoedema: lymphangiographic aspects. AB - We report a 5 1/2-year-old boy with juvenile rheumatoid arthritis (JRA) and lower limb lymphoedema. US, MRI and lymphangiography were performed. Based on the lymphangiographic study, we propose a pathogenesis based on obstruction of normal superficial lymphatic vessels in the affected limb. This is discussed with other pathogenetic factors proposed in the 16 previously reported cases of lymphoedema complicating JRA. PMID- 10382217 TI - MRI evaluation of infectious and non-infectious synovitis: preliminary studies in a rabbit model. AB - BACKGROUND: Literature on magnetic resonance imaging (MR) evaluation of inflammatory joint effusions is sparse. OBJECTIVE: To describe an animal model for studying infectious and non-infectious joint effusions with magnetic resonance imaging. MATERIALS AND METHODS: Ten rabbit knees with septic arthritis and four with talc synovitis were imaged with MR. Contralateral knees injected with saline served as controls. Fat saturation T2-weighted and gadolinium enhanced T1-weighted images were assessed for joint effusion, and periarticular and adjacent intraosseous increased signal or enhancement. Each knee was cultured and underwent pathologic examination. RESULTS: Both Staphylococcus aureus and talc produced effusions in all knees. The degree of periarticular signal and enhancement was greater in infected knees than talc-injected knees. No abnormal enhancement was seen within bone. Pathologic examination showed a greater degree of inflammation and joint destruction in the infected knees, but no evidence of osteomyelitis. CONCLUSION: A greater degree of abnormal signal and enhancement seen on MR suggests a more vigorous inflammatory process, as seen with septic arthritis. In spite of advanced septic arthritis, no enhancement was evident within bone, suggesting that enhancement within bone is not an expected finding in isolated septic arthritis and should raise concern for osteomyelitis. PMID- 10382218 TI - Metaphyseal anadysplasia in two sisters. AB - Metaphyseal anadysplasia is a rare form of metaphyseal chondrodysplasia with well defined radiological abnormalities. The prognosis is good as the natural course results in regression of the lesions with normal stature in adulthood. The few reported cases, exclusively in male children, indicated possible X-linked recessive transmission. The documentation of two affected sisters suggests genetic heterogeneity or another mode of inheritance. PMID- 10382219 TI - Bilateral smooth-muscle tumors of the adrenals in a child with AIDS. AB - We report a case of bilateral smooth muscle tumors (SMT) involving the adrenal glands in an 11-year-old female with acquired immunodeficiency syndrome (AIDS). The SMT of the right adrenal gland extended into the inferior vena cava, producing a tumor thrombus. PMID- 10382220 TI - Neuromuscular control: introduction and overview. AB - This paper introduces some basic concepts of the interdisciplinary field of neuromuscular control, without the intention to be complete. The complexity and multifaceted nature of neuromuscular control systems is briefly addressed. Principles of stability and planning of motion trajectories are discussed. Closed loop and open-loop control are considered, together with the inherent stability properties of muscles and the geometrical design of animal bodies. Various modelling approaches, as used by several authors in the Philosophical Transactions of the Royal Society of London, Series B, May 1999 issue, such as inverse and forward dynamics are outlined. An introductory overview is presented of the other contributions in that issue. PMID- 10382221 TI - Balancing on a narrow ridge: biomechanics and control. AB - The balance of standing humans is usually explained by the inverted pendulum model. The subject invokes a horizontal ground-reaction force in this model and controls it by changing the location of the centre of pressure under the foot or feet. In experiments I showed that humans are able to stand on a ridge of only a few millimetres wide on one foot for a few minutes. In the present paper I investigate whether the inverted pendulum model is able to explain this achievement. I found that the centre of mass of the subjects sways beyond the surface of support, rendering the inverted pendulum model inadequate. Using inverse simulations of the dynamics of the human body, I found that hip-joint moments of the stance leg are used to vary the horizontal component of the ground reaction force. This force brings the centre of mass back over the surface of support. The subjects generate moments of force at the hip-joint of the swing leg, at the shoulder-joints and at the neck. These moments work in conjunction with a hip strategy of the stance leg to limit the angular acceleration of the head-arms-trunk complex. The synchrony of the variation in moments suggests that subjects use a motor programme rather than long latency reflexes. PMID- 10382222 TI - Model-based development of neuroprosthesis for paraplegic patients. AB - In paraplegic patients with upper motor neuron lesions the signal path from the central nervous system to the muscles is interrupted. Functional electrical stimulation applied to the lower motor neurons can replace the lacking signals. A so-called neuroprosthesis may be used to restore motor function in paraplegic patients on the basis of functional electrical stimulation. However, the control of multiple joints is difficult due to the complexity, nonlinearity, and time variance of the system involved. Furthermore, effects such as muscle fatigue, spasticity, and limited force in the stimulated muscle further complicate the control task. Mathematical models of the human musculoskeletal system can support the development of neuroprosthesis. In this article a detailed overview of the existing work in the literature is given and two examples developed by the author are presented that give an insight into model-based development of neuroprosthesis for paraplegic patients. It is shown that modelling the musculoskeletal system can provide better understanding of muscular force production and movement coordination principles. Models can also be used to design and test stimulation patterns and feedback control strategies. Additionally, model components can be implemented in a controller to improve control performance. Eventually, the use of musculoskeletal models for neuroprosthesis design may help to avoid internal disturbances such as fatigue and optimize muscular force output. Furthermore, better controller quality can be obtained than in previous empirical approaches. In addition, the number of experimental tests to be performed with human subjects can be reduced. It is concluded that mathematical models play an increasing role in the development of reliable closed-loop controlled, lower extremity neuroprostheses. PMID- 10382223 TI - Simulations of neuromuscular control in lamprey swimming. AB - The neuronal generation of vertebrate locomotion has been extensively studied in the lamprey. Models at different levels of abstraction are being used to describe this system, from abstract nonlinear oscillators to interconnected model neurons comprising multiple compartments and a Hodgkin-Huxley representation of the most relevant ion channels. To study the role of sensory feedback by simulation, it eventually also becomes necessary to incorporate the mechanical movements in the models. By using simplifying models of muscle activation, body mechanics, counteracting water forces, and sensory feedback through stretch receptors and vestibular organs, we have been able to close the feedback loop to enable studies of the interaction between the neuronal and the mechanical systems. The neuromechanical simulations reveal that the currently known network is sufficient for generating a whole repertoire of swimming patterns. Swimming at different speeds and with different wavelengths, together with the performance of lateral turns can all be achieved by simply varying the brainstem input. The neuronal mechanisms behind pitch and roll manoeuvres are less clear. We have put forward a 'crossed-oscillators' hypothesis where partly separate dorsal and ventral circuits are postulated. Neuromechanical simulations of this system show that it is also capable of generating realistic pitch turns and rolls, and that vestibular signals can stabilize the posture during swimming. PMID- 10382224 TI - Haltere-mediated equilibrium reflexes of the fruit fly, Drosophila melanogaster. AB - Flies display a sophisticated suite of aerial behaviours that require rapid sensory-motor processing. Like all insects, flight control in flies is mediated in part by motion-sensitive visual interneurons that project to steering motor circuitry within the thorax. Flies, however, possess a unique flight control equilibrium sense that is encoded by mechanoreceptors at the base of the halteres, small dumb-bell-shaped organs derived through evolutionary transformation of the hind wings. To study the input of the haltere system onto the flight control system, I constructed a mechanically oscillating flight arena consisting of a cylindrical array of light-emitting diodes that generated the moving image of a 30 degrees vertical stripe. The arena provided closed-loop visual feedback to elicit fixation behaviour, an orientation response in which flies maintain the position of the stripe in the front portion of their visual field by actively adjusting their wing kinematics. While flies orientate towards the stripe, the entire arena was swung back and forth while an optoelectronic device recorded the compensatory changes in wing stroke amplitude and frequency. In order to reduce the background changes in stroke kinematics resulting from the animal's closed-loop visual fixation behaviour, the responses to eight identical mechanical rotations were averaged in each trial. The results indicate that flies possess a robust equilibrium reflex in which angular rotations of the body elicit compensatory changes in both the amplitude and stroke frequency of the wings. The results of uni- and bilateral ablation experiments demonstrate that the halteres are required for these stability reflexes. The results also confirm that halteres encode angular velocity of the body by detecting the Coriolis forces that result from the linear motion of the haltere within the rotating frame of reference of the fly's thorax. By rotating the flight arena at different orientations, it was possible to construct a complete directional tuning map of the haltere-mediated reflexes. The directional tuning of the reflex is quite linear such that the kinematic responses vary as simple trigonometric functions of stimulus orientation. The reflexes function primarily to stabilize pitch and yaw within the horizontal plane. PMID- 10382225 TI - The neuromuscular control of birdsong. AB - Birdsong requires complex learned motor skills involving the coordination of respiratory, vocal organ and craniomandibular muscle groups. Recent studies have added to our understanding of how these vocal subsystems function and interact during song production. The respiratory rhythm determines the temporal pattern of song. Sound is produced during expiration and each syllable is typically followed by a small inspiration, except at the highest syllable repetition rates when a pattern of pulsatile expiration is used. Both expiration and inspiration are active processes. The oscine vocal organ, the syrinx, contains two separate sound sources at the cranial end of each bronchus, each with independent motor control. Dorsal syringeal muscles regulate the timing of phonation by adducting the sound generating labia into the air stream. Ventral syringeal muscles have an important role in determining the fundamental frequency of the sound. Different species use the two sides of their vocal organ in different ways to achieve the particular acoustic properties of their song. Reversible paralysis of the vocal organ during song learning in young birds reveals that motor practice is particularly important in late plastic song around the time of song crystallization in order for normal adult song to develop. Even in adult crystallized song, expiratory muscles use sensory feedback to make compensatory adjustments to perturbations of respiratory pressure. The stereotyped beak movements that accompany song appear to have a role in suppressing harmonics, particularly at low frequencies. PMID- 10382226 TI - Neuromuscular control of prey capture in frogs. AB - While retaining a feeding apparatus that is surprisingly conservative morphologically, frogs as a group exhibit great variability in the biomechanics of tongue protraction during prey capture, which in turn is related to differences in neuromuscular control. In this paper, I address the following three questions. (1) How do frog tongues differ biomechanically? (2) What anatomical and physiological differences are responsible? (3) How is biomechanics related to mechanisms of neuromuscular control? Frog species use three non exclusive mechanisms to protract their tongues during feeding: (i) mechanical pulling, in which the tongue shortens as its muscles contract during protraction; (ii) inertial elongation, in which the tongue lengthens under inertial and muscular loading; and (iii) hydrostatic elongation, in which the tongue lengthens under constraints imposed by the constant volume of a muscular hydrostat. Major differences among these functional types include (i) the amount and orientation of collagen fibres associated with the tongue muscles and the mechanical properties that this connective tissue confers to the tongue as a whole; and (ii) the transfer of intertia from the opening jaws to the tongue, which probably involves a catch mechanism that increases the acceleration achieved during mouth opening. The mechanisms of tongue protraction differ in the types of neural mechanisms that are used to control tongue movements, particularly in the relative importance of feed-forward versus feedback control, in requirements for precise interjoint coordination, in the size and number of motor units, and in the afferent pathways that are involved in coordinating tongue and jaw movements. Evolution of biomechanics and neuromuscular control of frog tongues provides an example in which neuromuscular control is finely tuned to the biomechanical constraints and opportunities provided by differences in morphological design among species. PMID- 10382228 TI - Thoracoscopic procedure for intrathoracic diseases: current status in mainland China. AB - One hundred and five patients underwent a procedure of video-assisted thoracoscopic surgery (VATS) during a period of 5 years, in the Department of Cardiothoracic Surgery, Peking Union Medical College Hospital, Beijing, China. Among them, there were 36 cases suffering from spontaneous pneumothorax, 24 having a nodule of the lung, nine intrathoracic multiple nodules, 10 undefined pleural effusions, eight mediastinal mass, 15 interstitial fibrosis, two pericardial effusion, and one severe emphysema. In all patients of this series, the diagnosis was demonstrated and the suitable management was performed. PMID- 10382227 TI - Asthma and chronic obstructive airway diseases are associated with osteoporosis and fractures: a literature review. AB - The objective was to assess the association between asthma and/or chronic obstructive airway diseases (COAD), and osteoporosis, and appraise treatments of osteoporosis in these patients. MEDLINE and Excerpta Medica were searched for original research with control groups which tested the above association. One cohort and nine cross-section studies of bone density in patients with asthma and/or COAD were retrieved. These demonstrated clinically important bone density reductions of up to 29% in subjects, dependent upon daily oral corticosteroids, by a variety of measurement techniques, at various bone sites. Bone density reduction has also been less consistently reported in the absence of oral corticosteroids, suggesting that other factors including high-dose inhaled corticosteroids may have a role. Fracture studies. Three studies in oral corticosteroid-dependent asthmatics demonstrated a vertebral fracture prevalence up to 56%, and annual vertebral fracture incidence of up to 42%. The strength of the available evidence is limited, but suggests that patients with asthma and/or COAD are at increased risk of osteoporosis. The evidence of the association between osteoporosis and inhaled corticosteroids is much more limited than for oral corticosteroids. Bisphosphonates are promising agents to maintain and/or promote bone mass in this patient group. PMID- 10382229 TI - Perception of bronchodilation in subjects with asthma and smokers with airflow limitation. AB - Perception of the efficacy of bronchodilators in relieving airflow obstruction is a likely determinant of compliance with treatment in patients prescribed these drugs on an 'as needed' basis. This study aimed to determine whether bronchodilator-induced improvements in lung function are associated with improvements in breathing difficulty in subjects with asthma or smokers with airflow limitation. Twenty smokers with airflow limitation and 16 subjects with previously physician-diagnosed asthma received salbutamol (200 micrograms) and ipratropium bromide (80 micrograms). Spirometry and lung volumes were measured before and 40 min after bronchodilator. Subjects recorded changes in 'difficult breathing' on a visual analogue scale (VAS). After bronchodilator, forced expiratory volume in 1 s (FEV1) increased by 23.0 +/- 6.4% of baseline (mean +/- 95% CI) in smokers, and by 25.2 +/- 8.5% in the asthmatics, while VAS improved by 31 +/- 23% in smokers and 45 +/- 25% in asthmatics. However, these changes were not significantly correlated in either smokers (r = -0.04) or asthmatics (r = 0.15). In the asthmatic subjects, good perceivers (> 25% improvement in VAS) had greater improvements in lung volumes, as percentage predicted, than did poor perceivers. In the smokers, changes in lung function did not differ significantly between good and poor perceivers. Improvement in FEV1, as percentage predicted, was significantly correlated with improvement in VAS in good perceivers (asthma: r = 0.78, P < 0.01; smokers: r = 0.68, P < 0.05), but not in poor perceivers. Asthmatic subjects had good perception of improvements in lung function. However, in smokers with airflow limitation there is little correlation between improvement in lung function and sensation of breathing difficulty. In these subjects symptoms appear to be an unreliable guide for 'as needed' use of bronchodilators. PMID- 10382230 TI - Regulation of ventilation before and after sleep in patients with obstructive sleep apnoea. AB - The objective was to examine whether abnormal breathing during sleep may affect regulation of ventilation after awakening in patients with obstructive sleep apnoea (OSAS). In 19 patients with OSA and 12 normal subjects we examined ventilatory responses to hypoxia (HVR) and to hypercapnia (HCVR) before and after sleep (BS and AS), and compared the changes in ventilatory responses with respiratory events during sleep. In the OSA group, the values of resting ventilation were significantly smaller in AS than those in BS and end-tidal partial pressure of CO2 in arterial blood (Pco2) (PETCO2) rose significantly from BS to AS. The slopes of the HVR or HCVR did not differ between BS and AS. However, both the response lines shifted downward and minute ventilation (VE)80 (VE at arterial oxygen saturation (Sao2) of 80%) in HVR and VE60 (VE at PETCO2 of 60 mmHg) in HCVR decreased significantly from BS to AS. The percentage changes of VE80 and VE60 were significantly correlated with mean Sao2, total sleep time below Sao2 of 90% and lowest Sao2 during sleep. However, in normal subjects we observed no circadian variation in their ventilatory responses. These data support the hypothesis that repeated episodes of nocturnal hypoxia and hypercapnia may modify the regulation of ventilation after awakening in patients with OSA. PMID- 10382231 TI - An increase in hyaluronan by lung fibroblasts: a biomarker for intensity and activity of interstitial pulmonary fibrosis? AB - The purpose of the present study was to clarify the roles of hyaluronan (HA) production of lung fibroblasts in the pathogenesis of pulmonary fibrosis. Quantitative and comparative assessments of the HA levels in bronchoalveolar lavage fluid (BALF) and lung fibroblast-conditioned media (F-CM) were made at various stages during the development of bleomycin-induced pulmonary fibrosis in rats. In bleomycin-treated animals, the HA levels in F-CM increased significantly (P < 0.01) on day 1 after bleomycin treatment, peaked on day 3, and then gradually declined and returned to control values on days 14-28. The HA concentrations of BALF in the bleomycin group were significantly increased (P < 0.01) on day 3, were maximal on day 7, and thereafter gradually decreased, remaining significantly above normal values (P < 0.01) on day 14, but returning to control values by day 28. In the bleomycin group, the HA levels both in BALF and in F-CM were significantly correlated with the cell components in BALF and there was a significant correlation between the HA concentration in BALF and in the F-CM. Lung fibroblasts were activated and produced increased HA which resulted in excessive accumulation of HA in the lung in the early stage of pulmonary fibrosis; the increased HA synthesis of lung fibroblasts and enhanced HA concentrations of BALF might reflect the intensity of alveolitis and the disease activity. PMID- 10382232 TI - Effects of interleukin-1 beta on DNA synthesis in rat alveolar type II cells in primary culture. AB - Proliferation of alveolar type II cells is critical for restoration of the integrity of alveolar epithelium in alveolar injuries caused by a number of different aetiologies. Because effects of inflammatory cytokines on the proliferation of alveolar type II cells are not clear, we investigated the effects of interleukin-1 beta (IL-1 beta) on [3H]-thymidine incorporation into DNA in rat alveolar type II cells in primary culture. Interleukin-1 beta enhanced the [3H]-thymidine incorporation dose and time dependently. The increase of [3H] thymidine incorporation was observed in parallel with increased number of rat alveolar type II cells. The effect of IL-1 beta on [3H]-thymidine incorporation was additive to effects of growth factors which were known to act as mitogenic factors for type II cells. Anti-interleukin-1 beta antibody or IL-1 receptor antagonist partially inhibited the effects of IL-1 beta on [3H]-thymidine incorporation. Their combination completely inhibited the effects of IL-1 beta. In the absence of IL-1 beta, the combination inhibited the [3H]-thymidine incorporation to a level under that in the control. Isolated alveolar type II cells were immunocytochemically stained positive with anti-IL-1 beta antibody. Reverse transcriptase-polymerase chain reaction (RT-PCR) showed the presence of the mRNA for IL-1 beta in cultured alveolar type II cells. These results demonstrate that exogenous IL-1 beta stimulates DNA synthesis in alveolar type II cells and that the cells also produce IL-1 beta endogenously and suggest that endogenous IL-1 beta may mediate basal DNA synthesis of alveolar type II cells. PMID- 10382233 TI - The glandular component in congenital cystic adenomatoid malformation of the lung. AB - Although severe congenital cystic changes (CCC) of the lung may be fatal, less severe forms may regress or vanish spontaneously. With recent advances in sonography, asymptomatic CCC are increasingly found. Whether all CCC should be promptly excised, or not, is uncertain. Congenital cystic changes conceptually are bronchopulmonary foregut malformations (BPFM) with a predilection for malignant degeneration. Among all BPFM, congenital cystic adenomatoid malformation (CCAM) is most common. We therefore searched for evidence of early malignant transformation in five surgically excised and three autopsy lungs with CCAM. By light microscopy, CCAM resembled poorly formed and dilated bronchi, bronchioles and respiratory air spaces. Four lungs had multiple nodular aggregates of mucus producing cells; the glandular component (GC). By scanning electron microscopy, GC appeared as multiple micropolyps, resembling neuroepithelial bodies. By transmission electron microscopy, GC had a surface proliferation of cells with granules of the mucous type and a basal increase in cells with owl-eyed neuroendocrine granules. The glandular component in CCAM appeared similar to the mucous cells in hyperplastic polyps of the colon and a type of mucus producing bronchioloalveolar carcinoma. Our findings support the hypothesis that CCAM is caused by dysregulated paracrine growth of mature cells and extracellular matrices and that GC could have the potential for malignant transformation. Further clinical and laboratory studies of BPFM are needed for the appropriate management of congenital cystic changes. PMID- 10382234 TI - Measurement of exhaled nitric oxide concentration using nasal continuous negative pressure. AB - Contamination of nasal nitric oxide (NO) is a major obstacle when one needs to sample exhaled NO originating only from the lungs. To eliminate nasal NO, we used the nasal continuous negative pressure (nasal CNP) technique which, we verified, caused closure of the vellum. Exhaled gas was sampled from six healthy volunteers into fraction 1 (initially exhaled 200 mL) and fraction 2 (remainder of the gas) under three conditions; while subjects were wearing a noseclip, using nasal CNP at -5, -10 and -20 cm H2O, and under endotracheal intubation. Exhaled NO concentration ([NO]) obtained with nasal CNP was significantly lower, regardless of the pressure applied, than that measured with a noseclip, and was similar to and closely correlated to that obtained under intubation (F1, r = 0.90; F2, r = 0.88; P < 0.05). Real-time recorded [NO] obtained with nasal CNP of -5 cm H2O was again lower than that measured with a noseclip at any expiratory flow rate examined, indicating nasal NO contamination was eliminated irrespective of the flow rate. In conclusion, because a nasal CNP of -5 cm H2O was easily tolerated without any discomfort, this technique is a simple, easy and effective technique to eliminate nasal NO which should be widely applicable for the measurement of exhaled [NO]. PMID- 10382235 TI - Effect of early application of biphasic positive airway pressure on the outcome of extubation in ventilator weaning. AB - Extubation failure is significantly associated with increased morbidity and mortality in mechanically ventilated patients. In respiratory distress after extubation, non-invasive positive pressure ventilation (NIPPV) has been suggested to avoid the complications of invasive mechanical ventilation. The purpose of this study was to evaluate the effect of early application of NIPPV on extubation outcome. We conducted a prospective study in 93 extubated patients with a mean age of 72.7 +/- 14.7 years (range, 24-93). Elective extubation was performed in 56 patients and unplanned extubation occurred in 37 patients. After extubation, patients randomly received either biphasic positive airway pressure (BIPAP) therapy (n = 47) or unassisted oxygen therapy (n = 46). Non-invasive positive pressure ventilation was delivered via face mask in BIPAP group. Of the 93 extubated patients, 73 (78.5%) were successfully extubated, and 20 (21.5%) had to be re-intubated. There were no significant differences in age, sex, pre extubation blood gas data between re-intubated patients and those who were not re intubated. While seven of the 46 patients in the unassisted oxygen therapy group required re-intubation, 13 of the 47 BIPAP-treated patients also required re intubation. This difference was not statistically significant. The postextubation respiratory management, BIPAP or unassisted oxygen therapy, did not correlate with the extubation outcome, but the elective extubation had significantly better outcome than unplanned extubation. Patients with excessive bronchial secretions and intolerance to the equipment are poor candidates for NIPPV. We conclude that early application of BIPAP support did not predict a favourable extubation outcome. Our experience did not support the indiscriminate use of NIPPV to facilitate ventilator weaning. PMID- 10382237 TI - Community acquired parapneumonic thoracic empyema: predictors of outcome. AB - The objective of this study was to determine those factors which are predictive of outcome in cases of thoracic empyema that are solely community acquired. All patients admitted with a diagnosis of thoracic empyema in the Auckland region between 1993 and 1995 were reviewed retrospectively. Both clinical and radiological outcomes were determined at 3-6 month follow up. Radiological outcomes were scored on admission, discharge and follow up by blinded chest radiograph review based on lung field opacification, extent and thickness of pleura. Forty-six cases fulfilled inclusion criteria. Mean age was 56 +/- 21 years, 67% were male and there was no ethnic preponderance. Forty-six per cent of cases had specific underlying risk factors. Multi-loculated cases of empyema occurred in 63%, pleural aspirate was culture positive in 59% and Streptococcus milleri represented 42% of all positive cultures. Delay in appropriate management was 5.2 +/- 4.2 days. Fifty-six per cent of total procedures incorporated intrapleural streptokinase as an adjunct to drainage. The cure rate was 93.3% without requirement for surgical intervention. Two patients died early giving a 4.3% mortality rate. Radiological scores were 8.6 +/- 0.6 on admission, 6.3 +/- 1.5 on discharge and 2.5 +/- 2.5 at follow up. Improvement in radiological scores was statistically significant from admission to discharge and follow up (P < 0.001). The total duration of hospitalization was 22 +/- 10 days and by univariate analysis was predicted by: pleural sepsis not involving S. milleri (P = 0.001), specific predisposing factors (P = 0.02), and frank pus on pleural aspiration (P = 0.03). Total delay in management was suggestive in prolonging the duration of hospitalization (P = 0.07). Parapneumonic thoracic empyema is an important source of morbidity which is determined by both patient and management factors. Good results can be obtained by prompt medical therapy including tube drainage and intrapleural streptokinase without need for surgical intervention. PMID- 10382236 TI - A 21-aminosteroid, U-74006F, attenuates endotoxin-induced lung injury in awake sheep. AB - The purpose of the present study was to examine the efficacy of U-74006F, a 21 aminosteroid, on lung dysfunction induced by endotoxaemia in awake sheep with lung lymph fistula and haemodynamic monitoring. We measured pulmonary haemodynamics, lung lymph balance, circulating leucocyte count, arterial blood gas tensions, and levels of thromboxane (Tx) B2 and 6-keto-prostaglandin (PG) F1 alpha in plasma and lung lymph. We performed two experiments. In experiment 1 (n = 6), we intravenously infused Escherichia coli lipopolysaccharide endotoxin (1 microgram/kg) over 30 min and observed the parameters over 5 h. In experiment 2 (n = 6), we pretreated sheep with an intravenous bolus of U-74006F (2 mg/kg) 30 min before the infusion of endotoxin in the same manner of experiment 1, and continuously infused U-74006F (0.5 mg/kg per h) over 5 h after the bolus during the experiment. The U-74006F significantly suppressed the early pulmonary hypertension, the late increase in pulmonary permeability and the elevations of TxB2 and 6-keto-PGF1 alpha levels in plasma and lung lymph during the early period following endotoxaemia, although the compound did not change the time course of leucocytopenia and hypoxaemia. These findings suggest that the administration of U-74006F attenuates the lung dysfunction induced by endotoxaemia in awake sheep. PMID- 10382238 TI - Lung adenocarcinoma in lymphocytic interstitial pneumonitis associated with primary Sjogren's syndrome. AB - We experienced a rare case of lung adenocarcinoma associated with lymphocytic interstitial pneumonitis caused by primary Sjogren's syndrome. A 78-year-old woman was referred to our hospital because of progressive sicca syndrome and nodular opacities in the right lower lobe on chest radiograph. This patient was diagnosed as primary Sjogren's syndrome by a labial gland biopsy and classical clinical features including xerophthalmia, xerostomia and immunoserological findings. Pathological findings including immunohistochemical studies in a surgically resected lung revealed adenocarcinoma in lymphocytic interstitial pneumonitis associated with primary Sjogren's syndrome. There was no evidence of malignant lymphoma in the lymph nodes or resected lung tissue. Pulmonary involvement of Sjogren's syndrome is now regarded both clinically and histopathologically as a wide spectrum of lymphoproliferative disorders ranging from benign to malignant. However, lung cancer associated with primary Sjogren's syndrome, as in our case, has apparently not been reported previously. PMID- 10382239 TI - Syndrome of inappropriate secretion of antidiuretic hormone associated with amyotrophic lateral sclerosis in respiratory failure. AB - A 65-year-old man who had muscle weakness and dysarthria was admitted for investigation of motor neuron disease. He had lost 12 kg of weight in 6 months. Neurological findings disclosed upper and lower motor neuron disturbances with normal sensory nerve function, and needle electromyography showed a neurogenic pattern. Laboratory findings on admission demonstrated dilutional hyponatraemia due to an excessive secretion of antidiuretic hormone (ADH). Based on these findings, the patient was diagnosed as having the syndrome of inappropriate secretion of antidiuretic hormone (SIADH) associated with amyotrophic lateral sclerosis (ALS). During the night of first hospital day, the patient complained of severe dyspnoea, and mechanical ventilation was commenced. Following the mechanical ventilation, plasma ADH levels and serum sodium concentration were normalized. We propose that respiratory failure secondary to the atrophy of respiratory muscle might be responsible for the development of SIADH. PMID- 10382240 TI - Multifocal peripheral bronchial carcinoid tumour. AB - Peripheral bronchial carcinoids sometimes arise as single solid or nodular lesions in the periphery of the lung. We encountered a 74-year-old Japanese male with bronchial carcinoids that were widely disseminated throughout the lung parenchyma. Pulmonary function tests revealed mild airflow obstruction. A metastatic process was ruled out from primary malignancy and a histological examination revealed findings consistent with a peripheral bronchial carcinoid. Based on these findings, we concluded that this patient had a primary multifocal peripheral bronchial carcinoid. An immunohistochemical examination revealed immunoreactivity for chromogranin A and bombesin. The present case appears to be an unusual case of diffuse multifocal peripheral bronchial carcinoid, confirmed by immunohistochemistry. PMID- 10382241 TI - Fulminant psittacosis requiring mechanical ventilation and demonstrating serological cross-reactivity between Legionella longbeachae and Chlamydia psittaci. AB - Chlamydia psittaci infection typically causes a mild respiratory illness in humans. Severe respiratory failure requiring mechanical ventilation or intensive care therapy is an uncommon development. The aetiological agents causing severe community acquired pneumonia often remain undetermined. Serological tests may aid in diagnosis. We present two cases of fulminant psittacosis, one demonstrating early cross-reactivity with Legionella longbeachae. PMID- 10382242 TI - Posterolateral lumbar intertransverse process spine arthrodesis with recombinant human bone morphogenetic protein 2/hydroxyapatite-tricalcium phosphate after laminectomy in the nonhuman primate. AB - STUDY DESIGN: A nonhuman primate lumbar intertransverse process arthrodesis model was used to evaluate recombinant human bone morphogenetic protein 2 (rhBMP-2) in a hydroxyapatite-tricalcium phosphate (HA-TCP) carrier as a complete bone graft substitute. OBJECTIVES: To assess the ability of a ceramic material to serve as a carrier for various doses of rhBMP-2 as a bone graft substitute in a primate model of posterolateral intertransverse process spinal fusion after laminectomy. SUMMARY OF BACKGROUND DATA: The reported non-union rates for posterolateral lumbar spine fusion with autogenous iliac crest bone range from 5-35%. Recombinant human bone morphogenetic protein 2 has shown potential to serve as a bone graft substitute for posterolateral intertransverse process spine fusion. Although a resorbable collagen sponge was a suitable carrier in rabbits and dogs, it was too compressible for the paraspinal muscles in rhesus monkeys. This failure of the collagen carrier has prompted evaluation of the feasibility of an alternative carrier material and the required dose of rhBMP-2. METHODS: Twenty one adult rhesus monkeys underwent a laminectomy at L4-L5 followed by bilateral intertransverse process arthrodesis via the same midline incision (n = 16) or a minimally invasive video-assisted posterolateral approach (n = 5). Bone graft implants on each side consisted of either 5 cm3 of autogenous iliac crest bone or 60:40 HA-TCP blocks (1.2 x 0.5 x 3.7 cm) loaded with a solution containing 0, 6, 9, or 12 mg of rhBMP-2 per side. The monkeys were killed 24 weeks after surgery. Inspection, manual palpation, radiography, and histology were used to assess fusion and to detect any bony growth into the laminectomy defect. RESULTS: Fusion was not achieved in any of the monkeys treated with autogenous iliac crest bone graft. Both of the monkeys treated with the HA-TCP blocks with 0 mg rhBMP-2 achieved fusion. All 15 monkeys treated with the HA-TCP blocks and either of the three doses of rhBMP-2 achieved solid fusion. Two animals had extension of the fusion on one side because of malpositioned ceramic block. The results in animals fused via the minimally invasive video-assisted technique were the same as inthose fused with the open technique. Histologic analysis showed some ingrowth of bone into the ends but not-through the ceramic block in the absence of rhBMP 2. When the ceramic blocks were loaded with rhBMP-2 there was a dose-dependent increase in the amount and quality of bone throughout the ceramic carrier based on qualitative assessment. No significant bone encroachment on the exposed thecal sac through the laminectomy defect was observed in any of the monkeys. CONCLUSION: Hydroxyapatite-tricalcium phosphate proved to be a suitable carrier for rhBMP-2 in the posterolateral spine fusion model in rhesus monkeys. Even in the presence of a laminectomy defect, there was no evidence of bone induction outside the confines of the ceramic carrier. PMID- 10382243 TI - Reconstruction after multilevel corpectomy in the cervical spine. A sagittal plane biomechanical study. AB - STUDY DESIGN: An in vitro biomechanical study of reconstruction techniques used after multilevel cervical corpectomy. OBJECTIVES: To determine the biomechanical behavior of the cervical spine after a multilevel corpectomy and reconstruction with a strut graft and supplementation of the graft with anterior and posterior plates. SUMMARY OF BACKGROUND DATA: Reconstruction of the spine after multilevel corpectomy represents a significant challenge, with nonunion or graft dislodgment being relatively common. Anterior and posterior plate fixation have increased the possibilities for supplemental stabilization. Although some clinical studies have been performed to examine multilevel corpectomies reconstructed with plates, biomechanical studies are few and are limited to single-segment models. METHODS: Flexibility testing was performed on 11 intact cervical spine preparations. Flexibility testing was also conducted on the spine preparations after reconstruction with a strut graft, after supplementation of the graft with an anterior plate, and after supplementation of the graft with lateral mass plates. Physiologic moments were applied dynamically, and the three-dimensional motion of the specimen was recorded with stereophoto-grammetry. Failure testing was performed on the plated specimens in compression. Load displacement curves and failure modes were analyzed. RESULTS: The range of motion after reconstruction compared with the control was decreased 24% after strut grafting, 43% after application of an anterior plate, and 62% after application of posterior plates. Similarly, flexibility coefficients showed that the posterior plate technique was the least flexible, followed by the anterior plate technique, with the graft alone being the most flexible reconstruction construct. Load to initial failure tended to be higher in posterior than in anterior plate specimens, and screw pullout was the predominant failure mode. CONCLUSIONS: The application of plates to the cervical spine as an adjunct to bone graft may improve the surgeon's ability to stabilize the spine after multilevel corpectomy. Understanding the biomechanics of these devices and the potential mode of failure is important in their use. PMID- 10382244 TI - Internal spinal fixator stiffness has only a minor influence on stresses in the adjacent discs. AB - STUDY DESIGN: Stresses in vertebral endplates and discs were calculated using the three-dimensional nonlinear finite-element model of a lumbar spine with an internal spinal fixation device. OBJECTIVE: To determine the influence of fixator stiffness on stresses in the adjacent discs. SUMMARY OF BACKGROUND DATA: There are few computer models of the lumbar spine with a fixator. Most of these models neglect the muscle forces. Fixator stiffness is assumed to influence greatly the stresses in the adjacent discs. METHODS: Two three-dimensional nonlinear finite element models were used to determine stresses in the lumbar spine for standing and 60 degrees flexion of the upper body. One model had an internal spinal fixator, the other did not. In a parameter study, the diameters of the longitudinal rod of the fixator were assumed to be 3, 5, 7, and 10 mm. In the computer model, the forces of the trunk muscles were simulated. RESULTS: The diameter of the longitudinal rod strongly affected the fixator loads but hardly influenced the stresses in the vertebral endplates. The stresses in the bridged discs were strongly reduced. However, the internal fixator had only a minor influence on the stresses in the anulus fibrosus and the pressure in the nucleus pulposus of the adjacent discs. CONCLUSIONS: The stiffness of an internal spinal fixation device has only a minor influence on stresses in the adjacent discs. PMID- 10382245 TI - Surface strain distribution on thoracic and lumbar vertebrae under axial compression. The role in burst fractures. AB - STUDY DESIGN: The surface strain distribution on the thoracic and lumbar vertebrae during axial compressive loading was examined. OBJECTIVES: To examine the general mechanical behavior of the thoracic and lumbar vertebrae to evaluate their role in burst fractures. SUMMARY OF BACKGROUND DATA: Burst fractures are generally characterized by injury to the middle column and fracturing of the superior endplate. However, results in previous biomechanical investigations have not shown how these fractures are initiated during compression. METHODS: Twenty one thoracic and lumbar vertebrae (5 T10, 10 L1, and 6 L4) with upper and lower vertebrae were studied. Three-axis rosette strain gauges were cemented to 11 sites on the vertebral surface. An axial compressive load was applied, and the strain was measured in each specimen. The strain recorded by each rosette gauge was converted into a tensile, compressive, and shear component. RESULTS: The highest tensile and compressive strain was recorded at the base of the pedicle. Shear strain in the vertebral body was significantly higher than that in the lamina at all three spinal levels. At L1 and L4, the tensile strain at the superior vertebral rim was higher than that at the inferior rim. CONCLUSIONS: The high tensile and compressive strains found at the base of the pedicle of T10, L1, and L4 indicate that the base of the pedicle is the site of fracture initiation. The higher tensile strain at the superior vertebral rim of L1 and L4 supports the clinical observation of the thoracolumbar burst fractures. PMID- 10382246 TI - Atlas-axis facet asymmetry. Implications in manual palpation. AB - STUDY DESIGN: A basic study of six human cervical spines, documenting displacement with applied forces mimicking palpation. OBJECTIVES: To assess the issues of motion palpation of joint restrictions and the inferred link to disease. SUMMARY OF BACKGROUND DATA: Although several investigators have suggested that the issue of asymmetry and normal-abnormal function should be assessed, data are unavailable. METHODS: Atlas-axis specimens were harvested from six cadavers, cleaned of ligamentous and muscle tissue, and potted and secured with dental plaster. Forces (5-25 N) were applied along the mediolateral axis, and the corresponding displacement along three orthogonal axes were documented with infrared diodes and the Optotrak camera system (Northern Digital, Waterloo, Ontario, Canada). Specimen geometry and asymmetry were documented with plain radiographic film and a gimbal apparatus. RESULTS: Each of the six specimens displayed different behavior and differing degrees of asymmetry (e.g., facet inclination 17-35 degrees) so that each was analyzed as a case study. Asymmetrical and discontinuous force-displacement correlations were linked to anatomic asymmetry that appeared to be of natural occurrence. CONCLUSIONS: Asymmetrical joint geometry is common and causes asymmetrical joint dynamics. Thus, a clinician attempting to palpate vertebral motion would be misled by assuming that perceived restricted joint motion universally represented a finding potentially amenable to manipulation. For spine palpation to be a valid indicator for manipulation, the clinician applying it must first be able to differentiate between asymmetrical motion caused by vertebral fixation and that caused by asymmetrical joint anatomy. PMID- 10382247 TI - Magnetic resonance evaluation of the intervertebral disc, spinal ligaments, and spinal cord before and after closed traction reduction of cervical spine dislocations. AB - STUDY DESIGN: A prospective clinical study using magnetic resonance imaging of the cervical spine in a consecutive series of patients with cervical spine dislocations. OBJECTIVES: To determine the incidence of intervertebral disc herniations and injury to the spinal ligaments before and after awake closed traction reduction of cervical spine dislocations. SUMMARY OF BACKGROUND DATA: Prior series in which the prereduction imaging of disc herniations in the dislocated cervical spine are described have been anecdotal and have involved small numbers of patients. In addition, no uniform clinical criteria to define the presence of an intervertebral disc herniation in the dislocated cervical spine has been described. The incidence of disc herniations in the unreduced dislocated cervical spine is unknown. METHODS: Eleven consecutive patients with cervical spine dislocations who met the clinical criteria for an awake closed traction reduction had prereduction and postreduction magnetic resonance imaging. Using strict clinical criteria for the definition of an intervertebral disc herniation, the presence or absence of disc herniation, spinal ligament injury, and cord injury was determined. Neurologic status before, during, and after the closed reduction maneuver was documented. RESULTS: Disc herniations were identified in 2 of 11 patients before reduction. Awake closed traction reduction was successful in 9 of the 11 patients. Of the nine patients with a successful closed reduction, two had disc herniations before reduction, and five had disc herniations after reduction. No patient had neurologic worsening after attempted awake closed traction reduction. CONCLUSIONS: The process of closed traction reduction appears to increase the incidence of intervertebral disc herniations. The relation of these findings, however, to the neurologic safety of awake closed traction reduction remain unclear. PMID- 10382248 TI - Persistent osteopenia in adolescent idiopathic scoliosis. A longitudinal follow up study. AB - STUDY DESIGN: A follow-up study assessing the bone mineral dynamics in adolescent patients with idiopathic scoliosis and associated osteopenia. OBJECTIVES: To investigate whether osteopenia in patients with adolescent idiopathic scoliosis is a transient phenomenon or a persistent problem. SUMMARY OF BACKGROUND DATA: Investigators have suggested a significant correlation of osteopenia with adolescent idiopathic scoliosis. Because one half of the skeletal mass is acquired during the adolescent years, it is of importance to know whether the osteopenia is transient or persistent. METHODS: Using dual-energy x-ray absorptiometry, bone mineral density of bilateral proximal femurs was studied longitudinally in 70 healthy control subjects and 14 patients with adolescent idiopathic scoliosis with significant osteopenia more than 2 standard deviations below the mean normal value. RESULTS: The 14 girls with osteopenic adolescent idiopathic scoliosis who were followed up longitudinally for up to 3 years showed persistent and significantly lower bone mineral density when compared with normal age-, sex- and maturity-matched control subjects. CONCLUSIONS: Patients with adolescent idiopathic scoliosis are at increased risk of osteoporosis than are healthy adolescents. The lower rate of increase of bone mineral density in patients with adolescent idiopathic scoliosis who have low bone mineral density could predict a significantly lower peak bone mass in adulthood, with all the associated problems of osteoporosis. Further investigation is needed to define whether osteopenia-associated scoliosis has the same cause, pathogenetic mechanism, and risk of progression when compared with adolescent scoliosis without osteopenia. PMID- 10382249 TI - Rotations of a helix as a model for correction of the scoliotic spine. AB - STUDY DESIGN: A prospective study using intraoperative stereophotogrammetry to analyze helical motion of the spine during the correction of scoliosis. OBJECTIVE: To determine whether derotation systems rotate the scoliotic helix. SUMMARY OF BACKGROUND DATA: Scoliosis is a complex three-dimensional deformity that is difficult to visualize on standard radiographs. The use of stereophotogrammetry has allowed study of the deformity in three dimensions during surgical correction. METHODS: Thirty-five patients with right thoracic adolescent idiopathic scoliosis were studied using a stereophotogrammetry technique during surgical correction. Changes in vertebral unique rotations and spinal plane of maximum deformity were measured during three sequential stages of the surgery. RESULTS: The mean preoperative and postoperative Cobb angles were 58 degrees and 19 degrees, respectively. Most rotation occurred at the top and bottom vertebrae in the curve, averaging 10 degrees each but in opposite directions. The apical vertebra rotated the least in the structural curve, with an average rotation of 5 degrees. Much of the rotation occurred during the derotation maneuver with additional rotation occurring during the final distraction. The plane of maximum deformity changed from a mean of 50 degrees before instrumentation to 19 degrees at the end of the procedure. CONCLUSIONS: Multiple rotations of the scoliotic curve occur, and it can be shown when maximum rotations occur during surgery. Posterior derotational systems unwind or rotate the scoliotic helix and reposition the resultant sine wave toward the sagittal plane as described by the change in the plane of maximum deformity. PMID- 10382250 TI - Left thoracic curve patterns and their association with disease. AB - STUDY DESIGN: Analysis of clinical database material collected prospectively. OBJECTIVE: Examination of the association between lateralization of scoliotic curves and the existence of underlying disease. SUMMARY OF BACKGROUND DATA: It has been suggested that left thoracic scoliosis configurations are intrinsically pathologic, whereas the more usual right curve is, in a sense, "normal." METHODS: Research-based records were analyzed. Scoliosis configuration, patient gender, and diagnostic group were correlated. The results were interpreted within a biologic framework. RESULTS: Congenital and infantile idiopathic scoliosis showed a random right-left curve distribution. In older age groups, boys were more likely to have a left thoracic curve and to have underlying disease, but there was no association between the two. In girls, occurrence in childhood itself and left thoracic patterns within that group were associated with disease. Nevertheless, most curves were in the right thoracic pattern, regardless of cause. CONCLUSIONS: Although an association exists between left thoracic curves and disease, it is not strong enough to determine who should be intensively investigated, to the exclusion of other clinical findings, and it seems inappropriate for a different approach to be adopted on the basis of scoliosis pattern alone. Gender in males and age at occurrence in females are more important risk factors than in scoliosis configuration. All new cases of scoliosis treated by a physician warrant equally meticulous assessment, with more sophisticated investigative techniques where indicated by the complete clinical picture, of which curve lateralization is only a part. Biologic theories of left and right are more capable of dealing with the phenomenon of scoliosis lateralization than are simple mechanics or concepts of specific diseases. PMID- 10382251 TI - The association between static pelvic asymmetry and low back pain. AB - STUDY DESIGN: A cross-sectional case-control approach was used to estimate the association between low back pain of less than 12 months' duration and pelvic asymmetry among 21-50-year-old patients seeking physical therapy services. OBJECTIVE: To evaluate the premise that asymmetrical positioning of the innominates of the pelvis is a source of low back pain. SUMMARY OF BACKGROUND DATA: No published studies have been conducted to evaluate systematically the association between low back pain and pelvic asymmetry in a clinic-based sample. METHODS: Pelvic landmark data were obtained in 144 cases and 138 control subjects. The associations of low back pain with levels of pelvic asymmetry were estimated by use of odds ratios and 95% confidence intervals. Effect modification and confounding of the low back pain-pelvic asymmetry association by several factors was assessed and alternative asymmetry measures considered. RESULTS: Pelvic asymmetry was not positively associated with low back pain in any way that seemed clinically meaningful. Asymmetry of posterior superior iliac spine landmarks showed some evidence of a weak positive association with low back pain. CONCLUSIONS: In the absence of meaningful positive association between pelvic asymmetry and low back pain, evaluation and treatment strategies based on this premise should be questioned. PMID- 10382252 TI - The surgical treatment of far lateral L3-L4 and L4-L5 disc herniations. A modified technique and outcomes analysis of 25 patients. AB - STUDY DESIGN: A retrospective review of 25 patients who underwent a modified surgical procedure for the treatment of far lateral disc herniation. OBJECTIVES: To describe a modification of previous surgical techniques for the treatment of far lateral disc herniation and to review the outcomes in resolution of pain and improvement of functional status. SUMMARY OF BACKGROUND DATA: Lumbar disc herniations that occur far lateral to the intervertebral facet result in spinal nerve compression at L3-L4 and L4-L5. Previous surgical techniques have resulted in an increased risk of instability or continued postoperative back pain. METHODS: Twenty-five patients with far lateral disc herniation underwent surgery using an extreme lateral approach. There was no medial facetectomy or disruption of the pars interarticularis. The intertransverse ligament was released from the superior portion of the inferior transverse process, and the nerve was located before removal of the disc. Preoperative and postoperative visual analog pain scale and Oswestry functional status evaluation were reviewed along with complications to evaluate the efficacy of the surgery. RESULTS: No serious complications were noted, although transient neuropathic pain was common and was theorized to be caused by manipulation of the dorsal root ganglion during surgery. This pain was usually resolved within 4 to 6 weeks. The mean preoperative and postoperative visual analog scale scores were 7.7 and 4.2, respectively. The mean preoperative and postoperative Oswestry scores were 50.7% and 34.7%, respectively. Both of these improvements were statistically significant (P < 0.01). CONCLUSIONS: This far lateral approach allowed the nerve and far lateral disc herniations to be easily identified. Also, there was less blood loss and no medial facetectomy or disruption of the pars interarticularis. This is a safe, effective technique with no disruption of spinal stability. PMID- 10382253 TI - Spinal biomodeling. AB - STUDY DESIGN: A prospective trial of stereolithographic biomodeling in complex spinal surgery. OBJECTIVES: To investigate the use of stereolithographic biomodeling as an aid to complex spinal surgery. SUMMARY OF BACKGROUND DATA: Of the array of imaging methods available to assist the spinal surgeon, no single method provides a complete overview of the anatomy, although three-dimensional imaging has been shown to have advantages. METHODS: Stereolithographic biomodeling is a new technology that allows data from three-dimensional computed tomographic scans to be used to generate exact plastic replicas of anatomic structures. Five patients with complex deformities were selected: two children with congenital deformities, a patient with an osteoblastoma, a patient with basilar invagination caused by osteogenesis imperfecta, and a patient with a failed lumbar fusion. Computed tomographic scanning was performed and stereolithographic biomodels generated. The stereolithographic biomodels were used for patient education, operative planning, and surgical navigation. RESULTS: The surgeons reported that biomodeling was useful in complex spinal surgery and was an effective technology. Stereolithographic biomodels were found to be particularly useful in morphologic assessment, in the planning and rehearsal of surgery, for intraoperative navigation, and for informing patients about surgical procedures. CONCLUSIONS: Stereolithographic biomodeling allows imaging data to be displayed in a physical form. This intuitive medium may improve data display and allows surgical simulation on a proxy of the surgical site. Draw-backs of the technology were a minimum 24 hours' manufacturing time and the cost. PMID- 10382254 TI - Direct repair of spondylolysis without spondylolisthesis, using a rod-screw construct and bone grafting of the pars defect. AB - STUDY DESIGN: A retrospective study of patient outcome after pars repair using an original technique in patients with spondylolysis without spondylolisthesis and degenerative disk disease. OBJECTIVES: To assess the results of a new technique of internal fixation that avoids penetration of the spinal canal, temporary fixation of the lumbosacral junction, and postoperative bracing owing to stable instrumentation consisting of pedicle screws and a V-shaped rod resting against the inferior aspect on the spinous process and the posterior aspects of the laminas. SUMMARY OF BACKGROUND DATA: Previously described techniques for direct repair of a pars defect often require postoperative bracing and can require intracanal penetration of wires or hooks; screws passing directly through the defect, thereby lessening the bone surface available for bone grafting; and temporary fixation of the lumbosacral junction with a plate that must be removed. METHODS: Patients with painful pars defect not responding to conservative therapy and interfering with everyday life, sports, or work were considered to be eligible for direct repair of the spondylolysis rather than lumbosacral fusion, if there was no associated degenerative disk disease or spondylolisthesis. The surgical technique involves placement of screws on the pedicles of the involved vertebra and the fixation of the loose posterior arch with a solid rod bent in a V shape, taking purchase on the spinous process and laminas. A bone graft is placed under compression in the pars defect before the rod-screw construct is tightened. RESULTS: The first 10 patients who underwent this technique had an average follow-up of 35 months (range, 7 months to 5.3 years); mean age at operation was 26 years (range, 16-48 years). Six patients had an excellent result, returned to normal everyday life and work, and participated in sports when desired. The outcome in one patient was rated good and in one, fair. The procedure in one was considered a failure, although bone fusion seemed to have been obtained. Seven patients would recommend the operation, one would hesitate. No complications were encountered because of the specific design of the construct. CONCLUSIONS: This new technique offers the advantage of being easy and fast, it can be performed using a great number of available spinal instrumentations using rods and pedicle screws. There is no violation of the neural canal except in the case of a misplacement of pedicle screws. No postoperative brace was used, return to everyday life avoiding low back stress was immediate, and return to work or sports was possible 3 to 6 months after the procedure. This technique seems safe and effective but needs careful selection of patients, as do all other techniques for direct repair of pars interarticularis. PMID- 10382255 TI - Iatrogenic Mycobacterium infection after an epidural injection. AB - STUDY DESIGN: Case report. OBJECTIVES: Successful excision of the mass and identification of the causative agent by histologic and microbiologic studies. SUMMARY OF BACKGROUND DATA: Spinal pain, caused by an infective mass, developed in a 39-year-old man 3 months after an epidural injection for low back pain. METHODS: Exploratory surgery was performed to remove the mass, and histologic and microbiologic studies were conducted. RESULTS: The inflammatory mass was excised successfully, and several specimens were examined for bacteriologic presence. Histologic examination of the excised specimen showed chronic granulomatous inflammation, and subsequent microbiologic studies cultured an acid- and alcohol fast bacillus that was later identified as Mycobacterium fortuitum. CONCLUSION: A review of the literature shows that this is a particularly uncommon micro organism. PMID- 10382256 TI - The spinal manifestations of Stickler's syndrome. AB - STUDY DESIGN: A review of current knowledge, clinical publications, and recent concepts of the causes of Stickler's syndrome was correlated with a clinical review of the condition at the Children's Hospital of Eastern Ontario, Canada. OBJECTIVES: To acquaint orthopedic spine surgeons with the natural history, associated anomalies, and high incidence of spinal deformity and scoliosis in children with Stickler's syndrome. SUMMARY OF BACKGROUND DATA: Stickler's syndrome is a hereditary, progressive arthro-ophthalmopathy with an autosomal dominant inheritance pattern. The estimated incidence is 1 in 10,000 people, which is slightly more common than Marfan syndrome. METHODS: The experience with Stickler's syndrome was reviewed in seven children, 2-15 years of age, with particular attention to the spinal abnormalities secondary to the connective tissue dysplasia. RESULTS: Six of the children had kyphosis or scoliosis, and four had wedging or flattening of the vertebrae or platyspondylia. In general, the spinal changes became more prominent in the older children with Stickler's syndrome, with the spinal vertebrae affected by the generalized epiphyseal dysplasia. The treatment of scoliosis and kyphosis is no different in children with Stickler's syndrome. The most difficult aspect is in diagnosing the condition. CONCLUSIONS: The importance of recognizing the syndrome is to allow for the investigation and treatment of the many other associated connective tissue disorders associated with Stickler's syndrome, such as the high incidence of retinal detachment, mitral valve prolapse, and mandibular hypoplasia that may result in problems with anesthesia should the spine require surgical stabilization. PMID- 10382257 TI - Sciatica caused by cervical and thoracic spinal cord compression. AB - STUDY DESIGN: Two case reports of sciatica that was considered to be caused by cervical and thoracic spinal cord compression. OBJECTIVES: To point out that sciatica can be an initial major symptom in patients with cervical or thoracic spinal cord lesions. SUMMARY OF BACKGROUND DATA: Usually, tract pain caused by cord compression is considered to be diffuse and does not resemble sciatica. METHODS: Medical history, physical findings, and the results of imaging studies were reviewed in one case of cervical cord tumor and one case of thoracic kyphosis. RESULTS: In both cases, sciatica was the initial and major symptom. Imaging studies showed no lesion in the lumbar spine. In one patient, a cervical dumbbell tumor was found to compress the cervical cord, and in the other the spinal cord was severely compressed at the thoracic kyphosis. The sciatica disappeared immediately after decompression surgery in both cases. CONCLUSIONS: Leg pain resembling sciatica can be caused by cord compression at the cervical and thoracic level. Thoracic kyphosis may be a causative factor in sciatica, in addition to spinal cord tumor and disc herniation, which have been reported previously. PMID- 10382258 TI - Management of degenerative disc disease above an L5-S1 segment requiring arthrodesis. AB - Clear guidelines exist for treating spondylolisthetic deformity and instability. How the surgeon handles adjacent-level degenerative disease is not as well established. Because magnetic resonance imaging now provides us with far more information on the "health" of radiographically normal intervertebral discs, the treatment of dehydrated or degenerated discs adjacent to a fusion is becoming more problematic. In this discussion, two experts discuss their approach to symptomatic lumbosacral spondolisthesis accompanied by adjacent-level disc degeneration. Drs. Herkowitz and Abraham believe strongly that the adjacent segment should be left alone, whereas Dr. Albert recommends extending the fusion in many instances. PMID- 10382259 TI - Collagen crosslinks in human lumbar intervertebral disc aging. PMID- 10382261 TI - Comparison of the downstream pathways for degradation of nitrobenzene by Pseudomonas pseudoalcaligenes JS45 (2-aminophenol pathway) and by Comamonas sp. JS765 (catechol pathway). AB - Nitrobenzene is degraded by Pseudomonas pseudoalcaligenes JS45 via 2-aminophenol to 2-aminomuconic semialdehyde, which is further degraded to pyruvate and acetaldehyde. Comamonas sp. JS765 degrades nitrobenzene via catechol to 2 hydroxymuconic semialdehyde. In this study we examined and compared the late steps of degradation of nitrobenzene by these two microorganisms in order to reveal the biochemical relationships of the two pathways and to provide insight for further investigation of their evolutionary history. Experiments showed that 2-hydroxymuconate, the product of the dehydrogenation of 2-hydroxymuconic semialdehyde, was degraded to pyruvate and acetaldehyde by crude extracts of Comamonas sp. JS765, which indicated the operation of a classical catechol meta cleavage pathway. The semialdehyde dehydrogenases from Comamonas sp. JS765 and P. pseudoalcaligenes JS45 were able to metabolize both 2-amino- and 2-hydroxymuconic semialdehyde, with strong preference for the physiological substrate. 2 Aminomuconate was not a substrate for 4-oxalocrotonate decarboxylase from either bacterial strain. The close biochemical relationships among the classical catechol meta-cleavage pathway in Comamonas sp. JS765, 2-aminophenol meta cleavage pathways in P. pseudoalcaligenes JS45, and an alternative 2-aminophenol meta-cleavage pathway in Pseudomonas sp. AP-3 suggest a common evolutionary origin. PMID- 10382260 TI - Thiamin biosynthesis in prokaryotes. AB - Twelve genes involved in thiamin biosynthesis in prokaryotes have been identified and overexpressed. Of these, six are required for the thiazole biosynthesis (thiFSGH, thil, and dxs), one is involved in the pyrimidine biosynthesis (thiC), one is required for the linking of the thiazole and the pyrimidine (thiE), and four are kinase genes (thiD, thiM, thiL, and pdxK). The specific reactions catalyzed by ThiEF, Dxs, ThiDM, ThiL, and PdxK have been reconstituted in vitro and ThiS thiocarboxylate has been identified as the sulfur source. The X-ray structures of thiamin phosphate synthase and 5-hydroxyethyl-4-methylthiazole kinase have been completed. The genes coding for the thiamin transport system (thiBPQ) have also been identified. Remaining problems include the cloning and characterization of thiK (thiamin kinase) and the gene(s) involved in the regulation of thiamin biosynthesis. The specific reactions catalyzed by ThiC (pyrimidine formation), and ThiGH and ThiI (thiazole formation) have not yet been identified. PMID- 10382262 TI - Purification, characterization, and primary structure of a monofunctional catalase from Methanosarcina barkeri. AB - Methanosarcina barkeri is a strictly anaerobic, cytochrome-containing, methane forming archaeon. We report here that the microorganism contains a catalase, which was purified and characterized. The enzyme with an apparent molecular mass of 190 kDa was shown to be composed of four identical subunits of apparent molecular mass of 54 kDa. The heme-containing enzyme did not exhibit peroxidase activity, which indicates that it is a monofunctional catalase. This is substantiated by the primary structure, which is related to that of other monofunctional catalases rather than to that of bifunctional catalase peroxidases. The enzyme showed an [S]0.5V for H2O2 of 25 mM and an apparent Vmax of 200,000 U/mg; it was inhibited by azide ([I]0.5V = 1 microM) and cyanide ([I]0.5V = 5 microM) and inactivated by 1,2,4-aminotriazole. The activity was almost independent of the pH (between pH 4 and 10) and the temperature (between 15 degrees C and 55 degrees C). Comparison of the primary structure of monofunctional catalases revealed that the enzyme from M. barkeri is most closely related to the monofunctional catalase of Dictyostelium discoideum. PMID- 10382263 TI - Alternative schemes of butyrate production in Butyrivibrio fibrisolvens and their relationship to acetate utilization, lactate production, and phylogeny. AB - Butyrivibrio fibrisolvens strains D1 and A38 produced little lactate, but strain 49 converted as much as 75% of its glucose to lactate. Strain 49 had tenfold more lactate dehydrogenase activity than strains D1 or A38, this activity was stimulated by fructose 1,6-bisphosphate, and had a pH optimum of 6.25. A role for fructose 1,6-bisphosphate or pH regulation of lactate production in strain 49 was, however, contradicted by the observations that very low concentrations (< 0.2 mM) of fructose 1,6-bisphosphate gave maximal activity, and continuous cultures did not produce additional lactate when the pH was decreased. The lactate production of strain 49 was clearly inhibited by the presence of acetate in the growth medium. When strain 49 was supplemented with as little as 5 mM acetate, lactate production decreased dramatically, and most of the glucose was converted to butyrate. Strain 49 did not possess butyrate kinase activity, but it had a butyryl-CoA/acetate CoA transferase that converted butyryl-CoA directly to butyrate, using acetate as an acceptor. The transferase had a low affinity for acetate (K(m) of 5 mM), and this characteristic explained the acetate stimulation of growth and butyrate formation. Strains D1 and A38 had butyrate kinase but not butyryl-CoA/acetate CoA transferase, and it appeared that this difference could explain the lack of acetate stimulation and lactate production. Based on these results, it is unlikely that B. fibrisolvens would ever contribute significantly to the pool of ruminal lactate. Since relatives of strain 49 (strains Nor37, PI 7, VV1, and OB156, based on 16S rRNA sequence analysis) all had the same method of butyrate production, it appeared that butyryl-CoA/acetate CoA transferase might be a phylogenetic characteristic. We obtained a culture of strain B835 (NCDO 2398) that produced large amounts of lactate and had butyryl-CoA/acetate CoA transferase activity, but this strain had previously been grouped with strains A38 and D1 based on 16S rRNA sequence analysis. Our strain B835 had a 16S rRNA sequence unique from the one currently deposited in GenBank, and had high sequence similarity with strains 49 and Nor37 rather than with strains A38 or D1. PMID- 10382264 TI - Biochemical interaction of human neutrophil peptide-1 with Mycobacterium tuberculosis H37Ra. AB - The biochemical mechanism of action of human neutrophil peptide-1 (HNP-1) against Mycobacterium tuberculosis H37Ra was studied. Mycobacteria grown in the presence of a subinhibitory concentration (IC50) of HNP-1 showed a significant decrease in the biosynthesis of vital macromolecules, as shown by the incorporation of various radiolabeled precursors. Mycobacterial cells grown in the presence of HNP 1 exhibited surface changes, as was evident from the increased number of binding sites for L-anilinonaphthalene 8-sulfonate. Permeability studies carried out with spheroplasts showed a significantly high permeability to a fluorescent probe, N phenyl naphthylamine, in the presence of HNP-1. Significant changes in the cell wall and cell membrane were observed when HNP-1-grown cells were analysed by transmission electron microscopy. Our results suggest the mycobacterial cell wall/membrane to be the major target(s) of HNP-1. PMID- 10382265 TI - Identification of biosynthetic intermediates of the extracellular polysaccharide viilian in Lactococcus lactis subspecies cremoris SBT 0495. AB - Lactococcus lactis subspecies cremoris SBT 0495 produces the phosphopolysaccharide viilian, which consists of the repeating unit beta-D glucosyl-(1-->4)-(alpha-L-rhamnosyl-(1-->2))-(alpha-D-galac tose-1- phosphoryl-(- >3)-beta-galactosyl-(1-->4)-beta-D-glucose. A lipid extract was prepared from cells in the late exponential phase of growth and was hydrolyzed by hydrochloric acid under mild conditions to split lipid-linked intermediates in the extract into lipid and sugar moieties. Both moieties were purified by chromatographic techniques and were characterized to identify intermediates of the viilian biosynthetic pathway. A polyisoprenoid isolated from the chloroform-soluble fraction of the hydrolyzed lipid extract was identified by mass spectrometry as undecaprenol. Saccharides isolated from the water-soluble fraction of the hydrolyzed lipid extract by anion-exchange chromatography, were characterized by glycosidic linkage analysis to discriminate sugar moieties of intermediates of viilian biosynthesis from compounds liberated from cell wall components. Some oligosaccharide analogues contain a glycerol residue, suggesting that these are fragments of glycosylglycerides and/or lipoteichoic acid. Three fragments were identified to be glucose, galactosyl-(1-->4)-glucose, and rhamnosyl-(1-->2) galactosyl-(1-->4)-glucose, which are in agreement with the structure of the repeating unit of viilian. These saccharides most likely represent the first three steps of the sequential assembly of the repeating unit of the undecaprenol assembly. PMID- 10382266 TI - Transforming growth factor-beta and p-21: multiple molecular targets of decorin mediated suppression of neoplastic growth. AB - Decorin is a member of the small leucine-rich proteoglycan (SLRP) gene family that has recently become a focus in various areas of cancer research. The decorin protein consists of a core protein and a covalently linked glycosaminoglycan chain. Decorin binds to collagens type I, II and IV in vivo and promotes the formation of fibers with increased stability and changes in solubility. Further, the decorin core protein binds to growth factors, including transforming growth factor-beta (TGF-beta), to other intercellular matrix molecules such as fibronectin and thrombospondin, and to the decorin endocytosis receptor. Decorin may directly interfere with the cell cycle via the induction of p21WAF1/CIP1 (p21), a potent inhibitor of cyclin-dependent kinases (CDKs). Here, we discuss interactions of decorin with TGF-beta and with p21, both of which are relevant to carcinogenesis and tumor progression. TGF-beta is released by tumors of various histogenetic origins and promotes immunosuppression in the host and tumor immune escape by induction of growth arrest and apoptosis in immune cells, by downregulation of MHC II antigen expression and by changes in the cytokine release profiles of immune and tumor cells. Moreover, TGF-beta may modulate tumor growth in an autocrine and paracrine fashion, may mediate drug resistance, and may facilitate tumor angiogenesis. Decorin binds to TGF-beta, thus inhibiting its bioactivity, and is a direct or indirect negative modulator of TGF-beta synthesis. Ectopic expression of decorin results in the regression of rat C6 gliomas, an antineoplastic effect attributed to the reversal of TGF-beta-induced immunosuppression. On the other hand, de novo expression of decorin in colon cancer cells and some other tumor cells, even though not in glioma cells, results in an upregulation of p21 expression and a cell cycle arrest, presumably in a TGF beta-independent manner. Decorin expression is downregulated in many tumors but upregulated in the peritumoral stroma. By virtue of its growth regulatory and immunomodulatory properties, decorin promises to become a novel target for the experimental therapy of human cancers. PMID- 10382267 TI - Mechanisms of alpha-latrotoxin action. AB - The major component of black widow spider venom, alpha-latrotoxin, triggers massive exocytosis in a variety of neurosecretory cells. An important trigger for exocytosis is the calcium influx via alpha-latrotoxin-induced channels in biological membranes. However, this mechanism fails to explain exocytosis which occurred in the complete absence of extracellular calcium. Recently, sophisticated biochemical and molecular techniques have led to the discovery of novel alpha latrotoxin-binding membrane receptors: neurexins and latrophilin/CIRL (calcium-independent receptor for alpha-latrotoxin). Neurexins are single transmembrane proteins which bind to alpha-latrotoxin in a calcium-dependent manner and also interact with the synaptic vesicle protein, synaptotagmin. On the other hand, latrophilin is a seven-transmembrane protein and belongs to the family of G-protein-coupled receptors. The multitude of effects of alpha latrotoxin on exocytosis in different cell systems and the nature of its membrane targets are discussed in this article. The molecular details of how alpha latrotoxin binding is transduced eventually to exocytosis remain to be elucidated. PMID- 10382268 TI - Cytokine-induced conversion of mesencephalic-derived progenitor cells into dopamine neurons. AB - We have previously shown that a combination of the cytokines interleukin (IL)-1, IL-11, leukemia inhibitory factor (LIF), and glial cell line-derived neurotrophic factor (GDNF) can convert rat fetal (E14.5) mesencephalic progenitor cells into tyrosine hydroxylase (TH)-immunoreactive (ir) neurons in vitro. The experiments described here characterize the mesencephalic progenitor cells and their cytokine induced conversion into dopamine (DA) neurons. For all experiments, we used bromodeoxyuridine (BrdU)-ir cultures of (E14.5) mesencephalic progenitor cells that had been expanded at least 21 days. We first demonstrated that IL-1 induced DA neuron conversion in mesencephalic progenitors, but not in striatal progenitors (P < 0.001). Thus, these cells should be classified as lineage restricted progenitors, and not omnipotent stem cells. To further characterize cell populations in these cultures, we used monoclonal antibodies against Hu (an early marker for neurons), growth-associated protein (GAP)-43 (a marker for neuronal process extension), TH (a marker for DA neurons), and glial fibrillary acidic protein (GFAP, a marker for astrocytes). We assessed (E14.5) mesencephalic progenitor cell cultures (plated at 125,000 cells/cm2) incubated in the cytokine mixture (described above) or in complete media (CM, negative control). Following 7 days incubation, GFAP-positive cells formed a nearly confluent carpet in both types of cultures. However, numbers of Hu-ir and GAP-43-ir cells in the cytokine incubated cultures far exceeded those in CM-incubated controls (P = 0.0003, P = 0.0001, respectively), while numbers of TH-ir cells were 58-fold greater in the cytokine-incubated cultures versus CM-incubated controls. The TH phenotype persisted for 7 days following withdrawal of the differentiation media. Numerous double-labeled cells that were BrdU-ir and also TH-ir, or Hu-ir and also TH-ir, were observed in the cytokine-incubated cultures. These data suggest that cytokines "drive" the conversion of progenitor cells into DA neurons. PMID- 10382269 TI - Innervation of the rat pineal gland by pituitary adenylate cyclase-activating polypeptide (PACAP)-immunoreactive nerve fibres. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP)-immunoreactive nerve fibres were demonstrated in the rat pineal gland. These fibres entered the pineal gland through the conarian nerve at the distal tip of the gland. A high density of the fibres was observed in the capsule of the gland, from where the immunoreactive elements penetrated into the pineal perivascular spaces and parenchyma. The majority of PACAP-immunoreactive nerve fibres also contained calcitonin gene-related peptide (CGRP). Some PACAP-immunoreactive nerve fibres contained neuropeptide Y (NPY), but only occasionally was PACAP colocalized with vasoactive intestinal peptide (VIP). After removal of both superior cervical ganglia, a high number of PACAP-containing nerve fibres were still present in the gland. In the nervous system PACAP is present in two isoforms, PACAP-38 and PACAP 27. The concentration of PACAP-38 in the superficial pineal gland was determined by radioimmunoassay to be 20.4 pmol/g tissue at midday and 18.9 pmol/g tissue at midnight. The concentration of PACAP-27 was only about 3% of the concentration of PACAP-38. In summary, this study is the first demonstration of a PACAP-containing innervation of the rat pineal gland. The PACAP concentration in the pineal gland does not exhibit a day-night difference. The colocalization of PACAP with calcitonin gene-related peptide in the pincalopetal nerve fibres indicates that the majority of PACAP-immunoreactive nerve fibres might originate from the trigeminal ganglion. PMID- 10382270 TI - Ultrastructural localization of adhesion molecules in the healthy and inflamed choroid plexus of the mouse. AB - The functional expression of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1) and MAdCAM-1 in the choroid plexus is indicative of a role of this structure in the communication of the immune system with the central nervous system (CNS). In order to gain further insight into the possible functions of adhesion molecules expressed in the choroid plexus, we investigated the exact ultrastructural localization of VCAM-1, ICAM-1 and MAdCAM 1 on semithin and ultrathin cryosections of the choroid plexus of healthy mice and of mice suffering from experimental autoimmune encephalomyelitis (EAE). In the healthy choroid plexus VCAM-1 and ICAM-1, but not MAdCAM-1, could be detected on the apical surface of the choroid plexus epithelial cells. During EAE, immunoreactivity for VCAM-1 and ICAM-1 was dramatically increased. Additionally, apical expression of MAdCAM-1 was observed on individual choroid plexus epithelial cells during EAE. At the same time, VCAM-1, ICAM-1 or MAdCAM-1 were never present on the endothelial cells of the fenestrated capillaries within the choroid plexus. The polar expression of VCAM-1, ICAM-1 and MAdCAM-1 on the apical surface of choroid plexus epithelial cells, which form the blood-cerebrospinal fluid barrier, implies a previously unappreciated function of this barrier in the immunosurveillance of the CNS. PMID- 10382271 TI - Expression of neurotrophin receptors trkB and trkC and their ligands in rat adrenal gland and the intermediolateral column of the spinal cord. AB - Neurotrophins and their trk receptors constitute major classes of signaling molecules with important actions in the developing and adult nervous system. With regard to the sympathoadrenal cell lineage, which gives rise to sympathetic neurons and chromaffin cells, neurotrophin-3 (NT-3) and nerve growth factor (NGF) are thought to influence developing sympathetic neurons. Neurotrophin requirements of chromaffin cells of the adrenal medulla are less well understood than those for NGF. In order to provide the bases for understanding of putative functions of neurotrophins for the development and maintenance of chromaffin cells and their preganglionic innervation, in situ hybridization has been used to study the expression of brain-derived neurotrophic factor (BDNF) and NT-3, together with their cognate receptors trkB and trkC, in the adrenal gland and in the intermediolateral column (IML) of the spinal cord. BDNF is highly expressed in the embryonic adrenal cortex and later in cells of the cortical reticularis zone. Adrenal medullary chromaffin cells fail to express detectable levels of mRNAs for BDNF, NT-3, and their cognate receptors trkB and trkC. Neurons in the IML express BDNF and trkB, and low levels of NT-3 and trkC. Our data make it unlikely that BDNF and NT-3 serve as retrograde trophic factors for IML neurons but suggest roles of BDNF and NT-3 locally within the spinal cord and possibly for sensory nerves of the adrenal cortex. PMID- 10382272 TI - 2'-Deoxyadenosine causes cell death in embryonic chicken sympathetic ganglia and brain. AB - Previous work has shown that nucleosides produce apoptosis in sympathetic ganglion (SG) cells in vitro. The present study examined the effects of nucleosides on the development of the chick embryo in vivo with special attention to the SG and the optic tectum of the central nervous system. In the presence of an adenosine deaminase inhibitor, adenosine and 2'-deoxyadenosine (2'-dAdo) produced different toxicity patterns: both adenosine and 2'-dAdo were toxic to E3 embryos, but only 2'-dAdo was toxic at later stages (E6 1/2, E11). Dosage experiments on E6 1/2 embryos showed that adenosine was less toxic than 2'-dAdo and that 2'-dAdo in sublethal doses was teratogenic. We also examined the effects of 2'-dAdo on embryonic chicken SG and optic tectum in vivo to determine whether sublethal doses of 2'-dAdo produced cell death in these centers on E6 1/2 and 10. In the E6 1/2 SG, 2'-dAdo produced significant neuron loss (83%) and a decrease in SG volume (65%); however, at E10, there was only minor cell loss (7%) and no significant change in SG volume. In the optic tectum at E6 1/2, cell loss was confined mainly to the tectal ventricular zone, but there was little sign of cell loss in this organ at E10. Since cell production is vigorous in the SG and optic tectum at E6 1/2 but relatively low at E10, 2'-dAdo appears to work by stopping cell proliferation. The ineffectiveness of 2'-dAdo at E10 may result from the lethality of 2'-dAdo to the embryo at low concentrations (30 microM) in vivo, well below the apoptosis-inducing concentrations employed in vitro (100-300 microM). These data extend previous findings showing that purine and pyrimidine metabolism plays an important role in development. PMID- 10382273 TI - Cholinergic neurons of the pelvic autonomic ganglia and uterus of the female rat: distribution of axons and presence of muscarinic receptors. AB - Acetylcholine (ACh) stimulates contraction of the uterus and dilates the uterine arterial supply. Uterine cholinergic nerves arise from the paracervical ganglia and were, in the past, characterized based on acetylcholinesterase (AChE) histochemistry. However, the histochemical reaction for acetylcholinesterase provides only indirect evidence of acetylcholine location and is a nonspecific marker for cholinergic nerves. The present study: (1) reevaluated cholinergic neurons of the paracervical ganglia, (2) examined the cholinergic innervation of the uterus by using retrograde axonal tracing and antibodies against molecules specific to cholinergic neurons, choline acetyltransferase and the vesicular acetylcholine transporter, and (3) examined muscarinic receptors in the paracervical ganglia using autoradiography and a radiolabeled agonist. Most ganglionic neurons were choline acetyltransferase- and vesicular acetylcholine transporter-immunoreactive and were apposed by choline acetyltransferase/vesicular acetylcholine transporter-immunoreactive terminals. Retrograde tracing showed that some cholinergic neurons projected axons to the uterus. These nerves formed moderately dense plexuses in the myometrium, cervical smooth muscle and microarterial system of the uterine horns and cervix. Finally, the paracervical ganglia contain muscarinic receptors. These results clearly reveal the cholinergic innervation of the uterus and cervix, a source of these nerves, and demonstrate the muscarinic receptor content of the paracervical ganglia. Cholinergic nerves could play significant roles in the control of uterine myometrium and vasculature. PMID- 10382274 TI - Direct immunogold labeling of connexins and aquaporin-4 in freeze-fracture replicas of liver, brain, and spinal cord: factors limiting quantitative analysis. AB - Direct immunogold labeling and histological mapping of membrane proteins is demonstrated in Lexan-stabilized SDS-washed freeze-fracture replicas of complex tissues. Using rat brain and spinal cord as primary model systems and liver as a "control" tissue to identify preparation and labeling artifacts, we demonstrate the presence of connexin43 in freeze-fractured gap junctions of identified and mapped astrocytes and ependymocytes, and confirm the presence of connexin32 in freeze-fractured gap junctions in liver. In addition, the simultaneous double labeling of dissimilar proteins (connexin43 and aquaporin-4) is demonstrated in gap junctions and square arrays, respectively, in the plasma membranes of astrocytes and ependymocytes. Finally, double-side shadowing and conventional staining methods are used to reveal the extent of biological material present at the time of labeling and to investigate the dynamics of membrane solubilization, the primary artifacts that occur during labeling, and several factors limiting quantitative analysis. PMID- 10382275 TI - In vitro alteration of macrophage phenotype and function by serum lipids. AB - Diabetes (type I and type II) affects approximately 13 million people in the United States. Delayed and incomplete healing of wounds can be a major problem for diabetic patients. Macrophages are an important cell in the complex process of wound repair representing the major source of cytokines throughout the wound healing process. Cytokines mediate many of the cellular responses critical to timely wound repair. It has been suggested that diabetes impairs wound healing through disruption of local cytokine production. Our previous in vivo studies in rats demonstrated that diabetes-induced and diet-induced hyperlipidemia cause changes in macrophage phenotype and function (Iacopino 1995; Doxey et al. 1998), suggesting that alterations in macrophage cytokine profiles represent the cellular/molecular mechanism responsible for delayed wound healing. The purpose of this study was to investigate how monocyte maturation/differentiation and cytokine production were altered by serum lipids in an in vitro system using human cells. Commercially prepared purified human monocytes were cultured and exposed to serum lipids. Phenotypic analysis of differentiated macrophages was then performed by flow cytometry and fluorescent microscopy using surface antigens specific for various macrophage subsets. Selected cytokines in conditioned medium were assayed using commercial human enzyme-linked immunosorbent assay (ELISA) kits. We demonstrate that serum lipids cause an increase in monocytic differentiation leading to an inflammatory macrophage phenotype rather than a reparative/proliferative phenotype. We also show that serum lipids cause a generalized decrease in macrophage cytokine production using interleukin-1 beta (IL-1 beta), tumor necrosis factor alpha (TNF-alpha), platelet derived growth factor (PDGF), and transforming growth factor beta 1 (TGF-beta 1) as marker cytokines. Our present in vitro results using human cells confirm our previous in vivo studies in the rat and support the hypothesis that diabetes induced hyperlipidemia alters the monocyte differentiation process resulting in changes of macrophage subsets and cytokine release at the wound site, ultimately impairing the wound-healing process. PMID- 10382276 TI - Expression of vascular endothelial growth factor (VEGF) and its receptors during embryonic implantation in the golden hamster (Mesocricetus auratus). AB - Expression of vascular endothelial growth factor/vascular permeability factor (VEGF/VPF) and its receptors (flt-1 and flk-1) during the peri-implantation period (days 3, 4, 5, 6 and 7 post coitus) in the golden hamster was investigated by in situ hybridization, immunohistochemistry and the reverse transcription/polymerase chain reaction (RT-PCR). Three days after mating, in situ hybridization and immunohistochemical staining revealed weak VEGF expression only in the uterine epithelium; this expression was similar to that seen at oestrus. Flt-1 but no flk-1 immunoreactivity was observed. At day 4, the subepithelial stroma and embryo displayed immunoreactivity for VEGF and flt-1, whereas endothelial cells expressed both flt-1 and flk-1. At day 5, immunoreactivity for both VEGF and its receptors was detected in decidual cells and vascular endothelial cells. Only a few embryonic cells expressed VEGF mRNA but strong signals were noted in decidual cells. The patterns of VEGF and VEGF receptor expression were the same in the day-6 and day-7 embryos and decidua, except for an increase in intensity as development progressed. Based on these findings, we conclude that, in addition to its known actions on endometrial angiogenesis and tissue swelling, VEGF may also facilitate the proliferation and differentiation of the endometrium and help to sustain the avascular embryo during this early stage of development. PMID- 10382277 TI - Colocalization of BAX and BCL-2 in small intestine and kidney biopsies with different degrees of DNA fragmentation. AB - Morphological changes associated with apoptosis are closely correlated with the expression of specific proteins. However, the cause-effect relationships between the expression of these proteins and DNA degradation are barely known. For studying expression of apoptosis-related proteins in relation to different degrees of DNA fragmentation, the small intestine with its spatially organized continuum of proliferation, differentiation and death is a very useful preparation. Enterocytes towards the apex of the villi become increasingly susceptible to apoptosis. Here, this "apoptotic gradient" is used to demonstrate the presence of BAX and BCL-2 proteins in the cytoplasm of cells at the onset of apoptosis. In semithin serial sections of the small intestine, BAX, BCL-2 and DNA fragmentation were demonstrated. BAX and BCL-2 are always colocalized and only in cells with fragmented DNA. The gradient of BAX or BCL-2 staining is similar to the gradient of DNA fragmentation. Immunoreactivity for BCL-2 or BAX is most intense in cells that are prone to become apoptotic next in the course of cellular turnover but not in cells in an advanced apoptotic state, showing strongly condensed chromatin. When using the same technique on semithin sections of kidney biopsies, containing epithelia with low cellular turnover, we found DNA fragmentation mainly in the epithelial cells of the distal tubules. Similar to the situation in the enterocytes, BAX staining was confined to the cytoplasm of epithelial cells with a moderate degree of DNA fragmentation and reduced in epithelial cells with a high degree of DNA fragmentation. In contrast to the situation in the small intestine, very low levels of BCL-2 were found. The results suggest that expression of BCL-2 and BAX is related to cell damage as indicated by DNA fragmentation but not to advanced stages of cellular death, as indicated by chromatin condensation and cellular shrinkage. PMID- 10382278 TI - Metaphase-specific cell death in meiotic spermatocytes in mice. AB - Apoptosis of male germ cells is a widespread but little-understood phenomenon in many animal species. The elucidation of its mechanisms could be useful in the understanding of male infertility. We have examined the distribution of dying cells with the terminal transferase-mediated nick-end labeling (TUNEL) method and by an electron-microscopic procedure in the testes of 10 mouse strains, viz., C57BL/10 (B10), SL/NiA (SL), C57BL/6 (B6), C3H/He (C3H), BALB/c (BALB), DBA2 (DBA), CBA/J (CBA), MRL/MpJ(-)+/+ (M+), MRL/MpJ-lpr/lpr (lpr), and wild-type NJL mice (Mus musculus musculus). In the testes of the B10, NJL, SL, B6, C3H, BALB, DBA, and CBA mice, very few TUNEL-positive cells are distributed in the seminiferous tubules, whereas in the testes of the M+ and lpr mice, many TUNEL positive cells, which are restricted to stage XII seminiferous tubules, have been identified. The most important finding is that many metaphases of meiotic spermatocytes show a marked TUNEL-positive reaction. Some metaphases show apoptotic morphology electron-microscopically. These results suggest that the testes of MRL strains will provide a useful model for the study of the mechanism of metaphase-specific apoptosis in meiotic spermatocytes. PMID- 10382279 TI - Factors affecting proliferation and dedifferentiation of primary bovine oviduct epithelial cells in vitro. AB - The oviduct is the physiological site for key events in reproduction, such as capacitation of spermatozoa, fertilization and early embryonic development. Interactions between oviduct epithelial cells and gametes or embryos cannot sufficiently be studied in vivo. Therefore, model systems are needed which mimic in vivo conditions most closely. In this study we optimised the method for isolating bovine oviduct cells and compared different cell support materials as well as two culture systems (perfusion vs static culture) for their ability to maintain characteristic morphological and functional features of oviduct cells. Out of nine different cell support materials tested, cellulose nitrate (0.45 micron pore size) was the most suitable to maintain cells in a manner similar to freshly isolated oviduct epithelial cells. Comparing static vs perfusion culture by electron microscopy, morphological differences of the cells were insignificant in the first days of culture, while they became more evident after 8 days. The cells in the static system lost typical characteristics such as columnar shape, cilia and secretory protrusions, while these features were still present in perfusion culture. In addition, intense ciliogenesis and cytoplasmic organelles for protein synthesis were found under perfusion conditions. These findings were underlined by differences in expression of the oviduct-specific oestrus associated glycoprotein 85-97 kDa (GP 85-97) gene as revealed by semi quantitative reverse transcription-polymerase chain reaction (RT-PCR). The RNA levels of this specific gene were significantly higher in perfusion compared to the static culture system. Our data show clear advantages of perfusion vs static culture for primary bovine oviduct epithelial cells. PMID- 10382280 TI - Ultrastructure and composition of Call-Exner bodies in bovine follicles. AB - Call-Exner bodies are present in ovarian follicles of a range of species including human and rabbit, and in a range of human ovarian tumors. We have also found structures resembling Call-Exner bodies in bovine preantral and small antral follicles. Hematoxylin and eosin staining of single sections of bovine ovaries has shown that 30% of preantral follicles with more than one layer of granulosa cells and 45% of small (less than 650 microns) antral follicles have at least one Call-Exner body composed of a spherical eosinophilic region surrounded by a rosette of granulosa cells. Alcian blue stains the spherical eosinophilic region of the Call-Exner bodies. Electron microscopy has demonstrated that some Call-Exner bodies contain large aggregates of convoluted basal lamina, whereas others also contain regions of unassembled basal-lamina-like material. Individual chains of the basal lamina components type IV collagen (alpha 1 to alpha 5) and laminin (alpha 1, beta 2 and delta 1) have been immunolocalized to Call-Exner bodies in sections of fresh-frozen ovaries. Bovine Call-Exner bodies are presumably analogous to Call-Exner bodies in other species but are predominantly found in preantral and small antral follicles, rather than large antral follicles. With follicular development, the basal laminae of Call-Exner bodies change in their apparent ratio of type IV collagen to laminin, similar to changes observed in the follicular basal lamina, suggesting that these structures have a common cellular origin. PMID- 10382281 TI - Origin of neuronal-like receptors in Metazoa: cloning of a metabotropic glutamate/GABA-like receptor from the marine sponge Geodia cydonium. AB - To date, no conclusive evidence has been presented for the existence of neuronal like elements in Porifera (sponges). In the present study, isolated cells from the marine sponge Geodia cydonium are shown to react to the excitatory amino acid glutamate with an increase in the concentration of intracellular calcium [Ca2+]i. This effect can also be observed when the compounds L-quisqualic acid (L-QA) or L (+)-2-amino-4-phosphonobutyric acid (L-AP-4) are used. The effect of L-QA and L AP-4, both agonists for metabotropic glutamate receptors (mGluRs), can be abolished by the antagonist of group I mGluRs, (RS)-alpha-methyl-4 carboxyphenylglycine. These data suggest that sponge cells contain an mGluR-like protein. A cDNA encoding rat mGluR subtype 1 has been used to identify the complete nucleotide sequence of G. cydonium cDNA coding for a 528-amino-acid-long protein (59 kDa) that displays marked overall similarity to mGluRs and to gamma aminobutyric acid B receptors. The deduced sponge polypeptide, termed putative mGlu/GABA-like receptor, displays the highest similarity to the two families of metabotropic receptors within the transmembrane segment. The N-terminal part of the sponge sequence shows similarity to mGluR4 and mGluR5. These findings suggest that the earliest evolutionary metazoan phylum, the Porifera, possesses a sophisticated intercellular communication and signaling system, as seen in the neuronal network of higher Metazoa. PMID- 10382282 TI - Immunolocalization of Na+,K(+)-ATPase in the organs of the branchial cavity of the European lobster Homarus gammarus (Crustacea, Decapoda). AB - The localization of Na+,K(+)-ATPase in epithelia of the organs of the branchial cavity of Homarus gammarus exposed to seawater and dilute seawater was examined by immunofluorescence microscopy and immunogold electron microscopy with a monoclonal antibody IgG alpha 5 raised against the avian alpha-subunit of the Na ,K(+)-ATPase. In juveniles held in seawater, fluorescent staining was observed only in the epithelial cells of epipodites. In juveniles held in dilute seawater, heavier immunoreactivity was observed in the epithelial cells of epipodites, and positive immunostaining was also observed along the inner-side epithelial layer of the branchiostegites. No fluorescent staining was observed in the gill epithelia. At the ultrastructural level, the Na+,K(+)-ATPase was localized in the basolateral infolding systems of the epipodite and inner-side branchiostegite epithelia of juveniles held in dilute seawater, mostly along the basal lamina. The expression of Na+,K(+)-ATPase therefore differs within tissues of the branchial cavity and according to the external salinity. These and previous ultrastructural observations suggest that the epipodites, and to a lesser extent the inner-side epithelium of the branchiostegites, are involved in the slight hyper-regulation displayed by lobsters at low salinity. Enhanced Na+,K(+)-ATPase activity and de novo synthesis of Na+,K(+)-ATPase within the epipodite and branchiostegite epithelia may be key points enabling lobsters to adapt to low salinity environments. PMID- 10382283 TI - Localization of a visual Gq protein in the photoreceptors of a polychaete, Perinereis brevicirris (Annelida). AB - Heterotrimeric GTP-binding proteins (G proteins) play an important role in phototransduction. The presence of G-protein subclasses has been reported in photoreceptive membranes, e.g., the Gi subgroup (transducin) in vertebrate rods, and the Gq subgroup in the eyes of the Arthropoda and the Mollusca. We examined the immunoreactivity and distribution of a Gq homologue in the cerebral ocelli of Perinereis brevicirris (Polychaeta, Annelida) using an anti-GqC antibody raised against a conserved sequence at the C-terminal of the alpha-subunit of Gq (Gq alpha). The anti-GqC antibody labeled a 48-kDa band on the Western blot of proteins from the Perinereis ocelli. The anti-GtC antibody, which is raised against the C-terminal sequence of bovine transducin alpha-subunit (Gt-alpha), did not cross-react to the ocellar proteins of Perinereis. The rhabdomeric layers of the anterior and posterior ocelli were strongly labeled by anti-GqC on light microscopic immunohistology. Immunoelectron microscopy showed that the Gq molecules were specifically localized in the photoreceptive membrane of the rhabdomeric microvilli. These results suggest that the Gq protein plays a role in the phototransduction of the Perinereis ocelli. PMID- 10382284 TI - Nitric oxide synthase in the gastrointestinal tract of the estuarine crocodile, Crocodylus porosus. AB - The aim of this study was to investigate the distribution of nitric oxide synthase (NOS)-containing nerve cells in the gastrointestinal tract of a reptile and to compare it with the pattern in other vertebrate classes. In the estuarine crocodile, Crocodylus porosus, NOS-positive nerve cell bodies and fibres were found in all regions of the gut examined. Most myenteric microganglia contained one or several NOS-immunoreactive neurons together with unlabelled neurons. The majority of the neurons were multipolar, ranging from 10 to 25 microns in diameter. Both the circular and the longitudinal muscle layers were innervated by NOS-immunoreactive nerve fibres, which mostly ran parallel to the muscle fibres. In addition, small blood vessels in the submucosa and on the serosal surface of the gut were innervated by NOS-immunoreactive fibres. Double labelling with antisera to NOS and vasoactive intestinal peptide (VIP) revealed three neuronal subpopulations. A small proportion of the NOS-immunoreactive cells also contained immunoreactivity to VIP while a majority of the VIP-immunoreactive cells were NOS immunoreactive. There were more nerve fibres showing VIP immunoreactivity than fibres with NOS immunoreactivity, although most of the latter also contained immunoreactivity to VIP. VIP-immunoreactive fibres often surrounded the NOS immunoreactive nerve cells. These results suggest that neuronally released nitric oxide is likely to be involved in the control of gastrointestinal motility in the crocodile as in most other vertebrate species. PMID- 10382285 TI - The surface-connected canalicular system in the sinus endothelial cells of rat spleen. AB - The existence of a surface-connected canalicular system in the splenic sinus endothelial cells of the rat has been demonstrated by transmission electron microscopy with lanthanum nitrate acting as a tracer for the extracellular space. In addition, the three-dimensional arrangement of the canaliculi has been revealed by computer-aided reconstruction. The surface-connected canalicular system of the sinus endothelial cells consists of slender canaliculi that are branched, anastomosed, and that show continuity with the plasma membrane. They twist in and out among the organelles and are often found in close apposition to the spherical invaginations of the plasma membrane and run alongside them. Canaliculi which are not infiltrated by lanthanum nitrate take the form of electron-lucent tubules and are accompanied by numerous spherical invaginations of the plasma membrane. From a computer-aided reconstruction, the canaliculi, which invaginate from various sites of the plasma membrane, have been found to be continuous with each other and to penetrate to the surface of the sinus endothelial cell; they also branch and anastomose to form a complex network in the cytoplasm. Although the surface-connected canalicular system in blood platelets and thrombocytes is believed to function as the main route for the discharge of granules and the uptake of foreign materials and also to take part in the storage and transport of calcium, it is unclear at present whether the network of the surface-connected canalicular system in splenic sinus endothelial cells has any physiological significance. PMID- 10382286 TI - Identification of microsatellite sequences in Vitis riparia and their applicability for genotyping of different Vitis species. AB - A Vitis riparia genomic library was screened for the presence of (GA)n simple sequence repeats (SSR) and 18 primer pairs yielding amplification products of the expected size were designed. Heterologous amplification with the primer pairs in related species (V. rupestris, V. berlandieri, V. labrusca, V. cinerea, V. aestivalis, V. vinifera, and interspecific hybrids) was successful in most primer species combinations. Therefore, the new markers are applicable to the genotyping of a range of Vitis species. Variations in the SSR flanking sequence were detected between and within the species. The degree of polymorphism and performance of the markers were determined in up to 120 individuals of V. vinifera. Four of fifteen alleles per locus were detected and expected heterozygosity ranged between 0.37 and 0.88. Null alleles were shown to be present at two loci by a lack of heterozygous individuals and by transmission of the null alleles in a controlled cross. Regular Mendelian inheritance is indicated for all but one loci by a preliminary segregation analysis in 36 offspring. Thirteen of the markers were found suitable for the genotyping of grapevines (V. vinifera). PMID- 10382288 TI - Development of amplified consensus genetic markers (ACGM) in Brassica napus from Arabidopsis thaliana sequences of known biological function. AB - A method for the development of consensus genetic markers between species of the same taxonomic family is described in this paper. It is based on the conservation of the peptide sequences and on the potential polymorphism within non-coding sequences. Six loci sequenced from Arabidopsis thaliana, AG, LFY3, AP3, FAD7, FAD3, and ADH, were analysed for one ecotype of A. thaliana, four lines of Brassica napus, and one line for each parental species, Brassica oleracea and Brassica rapa. Positive amplifications with the degenerate primers showed one band for A. thaliana, two to four bands in rapeseed, and one to two bands in the parental species. Direct sequencing of the PCR products confirms their peptide similarity with the "mother" sequence. By comparison of intron sequences, the correspondence between each rapeseed gene and its homologue in one of the parental species can be determined without ambiguity. Another important result is the presence of a polymorphism inside these fragments between the rapeseed lines. This variability could generally be detected by differences of electrophoretic migration on long non-denaturing polyacrylamide gels. This method enables a quick and easy shuttle between A. thaliana and Brassica species without cloning. PMID- 10382287 TI - The role of rDNA genes in X chromosome association in the aphid Acyrthosiphon pisum. AB - Silver staining of mitotic metaphases of the aphid A. pisum reveals the presence of argentophilic bridges connecting the two X chromosomes. The presence of nucleolar material connecting sex chromosomes seems to be quite a common phenomenon in organisms belonging to very different phyla, and suggests a role of nucleolar proteins in chromosome association and disjunction. In somatic cells of A. pisum, bridges connecting X chromosomes are detectable not only after silver staining but also after CMA3 staining. This finding suggests that GC rich DNA is involved in this type of association. Molecular analysis of rDNA intergenic spacers shows several 247 bp repeats containing short sequences having a high level of homology with the chi sequence of Escherichia coli and with the consensus core region of human hypervariable minisatellites. Moreover, each 247 bp repeat presents a perfect copy of a promoter sequence for polymerase I. These aphid repeats show structural homologies with a 240 bp repeat, which is considered to be responsible for sex chromosome pairing in Drosophila, not only in view of their common presence within rDNA spacers but also for their length and structure. The presence of chi sequences in the IGS of A. pisum, by promoting unequal crossing-over between rDNA genes, could thus give rise to the nucleolar organizing region (NOR) heteromorphism described in different aphid species. Although X pairing at NORs is fundamental in aphid male determination, the presence of heteromorphism of rDNA genes does not inhibit male determination in the A. pisum clone utilized for our experiments. PMID- 10382289 TI - Rediscovery and further characterization of the aeroplane (ae) wing posture mutation in Drosophila melanogaster. AB - In 1931, Theodore Quelprud characterized a novel spontaneous mutation in Drosophila melanogaster, which was named aeroplane (ae) based on its abnormal wing posture. Although the characterization of the original ae locus was minimal, it is very likely that another allele of this extinct mutation has now been identified. aeroplane-like (ae-l) was isolated as a by-product of a transformation experiment. The apparent wing paralysis is not caused by any obvious abnormalities in the thorax, wing, indirect flight muscles or direct flight muscles. Classical genetic complementation analyses of ae-l with other genes in the region suggest that it represents an allele of a novel locus. Unexpectedly, a molecular examination revealed that the physical lesion identified in the ae-l mutant is exceptionally close to the homeotic gene teashirt (tsh) and, indeed, may represent an unusual allele of teashirt. PMID- 10382290 TI - Genetic diversity in Elymus caninus as revealed by isozyme, RAPD, and microsatellite markers. AB - Genetic diversity of 33 Elymus caninus accessions was investigated using isozyme, RAPD, and microsatellite markers. The three assays differed in the amount of polymorphism detected. Microsatellites detected the highest polymorphism. Six microsatellite primer pairs generated a total of 74 polymorphic bands (alleles), with an average of 15.7 bands per primer pair. Three genetic similarity matrices were estimated based on band presence or absence. Genetic diversity trees (dendrograms) were derived from each marker technique, and compared using Mantel's test. The correlation coefficients were 0.204, 0.267, and 0.164 between isozyme and RAPD distance matrices, RAPD and microsatellite distance matrices, and between isozyme and microsatellite distance matrices, respectively. The three methodologies gave differing views of the amount of variation present but all showed a high level of genetic variation in E. caninus. The following points may be drawn from this study whether based on RAPD, microsatellite, or isozyme data: (i) The Icelandic populations are consistently revealed by the three dendrograms. The congruence of the discrimination of this accession group by RAPD, microsatellite, and isozyme markers suggests that geographic isolation strongly influenced the evolution of the populations; (ii) The degree of genetic variation within accessions was notably great; and (iii) The DNA-based markers will be the more useful ones in detecting genetic diversity in closely related accessions. In addition, a dendrogram, which took into account all fragments produced by isozymes, RAPDs, and microsatellites, reflected better the relationships than did dendrograms based on only one type of marker. PMID- 10382291 TI - A subtelomeric satellite DNA family isolated from the genome of the dioecious plant Silene latifolia. AB - In an ongoing effort to trace the evolution of the sex chromosomes of Silene latifolia, we have searched for the existence of repetitive sequences specific to these chromosomes in the genome of this species by direct isolation from low melting agarose gels of satellite DNA bands generated by digestion with restriction enzymes. Five monomeric units belonging to a highly repetitive family isolated from Silene latifolia, the SacI family, have been cloned and characterized. The consensus sequence of the repetitive units is 313 bp in length (however, high variability exists for monomer length variants) and 52.9% in AT. Repeating units are tandemly arranged at the subtelomeric regions of the chromosomes in this species. The sequence does not possess direct or inverted sequences of significant length, but short direct repeats are scattered throughout the monomer sequence. Several short sequence motives resemble degenerate monomers of the telomere repeat sequence of plants (TTTAGGG), confirming a tight association between this subtelomeric satellite DNA and the telomere repeats. Our approach in this work confirms that SacI satellite DNA sequences are among the most abundant in the genome of S. latifolia and, on the other hand, that satellite DNA sequences specific of sex chromosomes are absent in this species. This agrees with a sex determination system less cytogenetically diverged from a bisexual state than the system present in other plant species, such as R. acetosa, or at least a lesser degree of differentiation between the sex chromosomes of S. latifolia and the autosomes. PMID- 10382292 TI - A twenty strain survey and backcross localization of the erythrocytic GTP concentration determining locus Gtpc on mouse chromosome 9. AB - Erythrocyte nucleotide concentrations were surveyed among 20 inbred strains of mice in order to further assess the variability in GTP concentration. There was no significant difference in erythrocytic ATP concentration (Scheffe's test at P = 0.01), 678-1154 nmol/mL packed cells, among the strains surveyed. Two groups were distinguishable with respect to erythrocytic GTP concentration, 8 strains having high GTP, 215 +/- 44 nmole/mL packed cells, and 12 strains having low GTP, 34 +/- 12 nmole/mL packed cells. The erythrocytic GTP concentration determining trait Gtpc was previously shown to be linked to transferrin, Trf, on chromosome 9. Analysis of 232 [(B6 x WB) F1 x B6] backcross individuals for Gtpc and 8 microsatellite markers restricted the localization of Gtpc to a 5.6 +/- 2.1 cM region. The gene order and genetic distances in cM +/- SE are: (D9Mit14) 0.4 +/- 0.4 (D9Mit24) 1.7 +/- 0.8 (Gtpc, D9Mit51, D9Mit116, D9Mit212) 3.9 +/- 1.3 (D9Mit200) 3.0 +/- 1.1 (D9Mit20) 7.8 +/- 1.8 (D9Mit18). The GTP concentration determining trait appears to be a property of erythrocytes as no differences were observed for GTP/ATP ratios of brain, kidney, liver, and tongue from a low GTP strain, C3H/HeHa x Pgk-la and a high GTP strain, C57BL/6J. PMID- 10382293 TI - Generation and chromosome mapping of expressed sequence tags (ESTs) from a human infant thymus. AB - In an effort to identify novel genes that are expressed differentially in an infant thymus, we constructed an oligo-d(T) primed cDNA library from a human infant thymus followed by single-run partial sequencing to generate expressed sequence tags (ESTs). Characterization of more than 1400 sequences enabled us to convert human thymus transcripts into 1223 useful ESTs. These ESTs consisted of 613 (50.1%) showing homology to known human genes, 51 (4.2%) matching to genes from other species, 289 (23.6%) matching ESTs of unknown functions, and 182 (14.9%) being novel transcripts. The expression profile of an infant thymus features a high number of genes related to cell division-DNA synthesis and gene protein expression, indicating the active growth stage of an infant thymus. To identify the chromosomal localization of 43 thymus ESTs, PCR-based mapping was performed using a human-rodent somatic cell hybrid or radiation hybrid mapping panel. The results indicated that several novel genes were determined to be located in the vicinity of previously mapped disease loci; histidinemia loci, plasminogen Tochigi disease loci, Ehlers-Danlos syndrome, hypertriglyceridemia, thyroid resistance locus, ocular albinism, galactosemia, porphyria variegata, Charcot-Marie-tooth disease, FEOM (fibrosis of extraocular muscles), Prader-Willi syndrome. PMID- 10382294 TI - Diversity among Cynodon accessions and taxa based on DNA amplification fingerprinting. AB - The genus Cynodon (Gramineae), comprised of 9 species, is geographically widely distributed and genetically diverse. Information on the amounts of molecular genetic variation among and within Cynodon taxa is needed to enhance understanding of phylogenetic relations and facilitate germplasm management and breeding improvement efforts. Genetic relatedness among 62 Cynodon accessions, representing eight species, was assessed using DNA amplification fingerprinting (DAF). Ten 8-mer oligonucleotides were used to amplify specific Cynodon genomic sequences. The DNA amplification products of individual accessions were scored for presence (1) or absence (0) of bands. Similarity matrices were developed and the accessions were grouped by cluster (UPGMA) and principal coordinate analysis. Analyses were conducted within ploidy level (2x = 18 and 4x = 36) and over ploidy levels. Each primer revealed polymorphic loci among accessions within species. Of 539 loci (bands) scored, 496 (92%) were polymorphic. Cynodon arcuatus was clearly separated from other species by numerous monomorphic bands. The strongest species similarities were between C. aethiopicus and C. arcuatus, C. transvaalensis and C. plectostachyus, and C. incompletus and C. nlemfuensis. Intraspecific variation was least for C. aethiopicus, C. arcuatus, and C. transvaalensis, and greatest for C. dactylon. Accessions of like taxonomic classification were generally clustered, except the cosmopolitan C. dactylon var. dactylon and C. dactylon var. afganicus. Within taxa, accessions differing in chromosome number clustered in all instances indicating the 2x and 4x forms to be closely related. Little, if any, relationship was found between relatedness as indicated by the DAF profiles and previous estimates of hybridization potential between the different taxa. PMID- 10382295 TI - Genetic variation within and relatedness among wood and plains bison populations. AB - There are two recognized subspecies of bison, wood (Bison bison athabascae) and plains (Bison bison bison) bison. The establishment of most bison populations from a small number of individuals has raised concerns about their genetic variation. To this end, 11 bison populations were surveyed with 11 microsatellite loci in order to calculate genetic variation and genetic distances. Mean number of alleles ranged between 3.18 at Antelope Island State Park (Utah) and 6.55 at Wood Buffalo National Park (Alberta and Northwest Territories). Mean heterozygosity ranged from 0.295 at Antelope Island State Park to 0.669 at Custer State Park (South Dakota). The amount of genetic variability present in the bison populations as measured by mean number of alleles and overall probability of identity was found to correlate with the number of founders for all sampled populations. The G-test for heterogeneity revealed some evidence for the existence of subpopulations at Wood Buffalo National Park, however very small genetic distances between these subpopulations suggest that nuclear material from the plains bison introduced into Wood Buffalo National Park has diffused throughout the park. Genetic distances between the sampled populations were generally larger between than within the two bison subspecies. PMID- 10382296 TI - Molecular cytogenetic analysis of Aegilops cylindrica host. AB - The genomic constitution of Aegilops cylindrica Host (2n = 4x = 28, DcDcCcCc) was analyzed by C-banding, genomic in situ hybridization (GISH), and fluorescence in situ hybridization (FISH) using the DNA clones pSc119, pAs1, pTa71, and pTA794. The C-banding patterns of the Dc- and Cc-genome chromosomes of Ae. cylindrica are similar to those of D-and C-genome chromosomes of the diploid progenitor species Ae. tauschii Coss. and Ae. caudata L., respectively. These similarities permitted the genome allocation and identification of the homoeologous relationships of the Ae. cylindrica chromosomes. FISH analysis detected one major 18S-5.8S-25S rDNA locus in the short arm of chromosome 1Cc. Minor 18S-5.8S-25S rDNA loci were mapped in the short arms of 5Dc and 5Cc. 5S rDNA loci were identified in the short arm of chromosomes 1Cc, 5Dc, 5Cc, and 1Dc. GISH analysis detected intergenomic translocation in three of the five Ae. cylindrica accessions. The breakpoints in all translocations were non-centromeric with similar-sized segment exchanges. PMID- 10382298 TI - Genome structure and evolution in the allohexaploid weed Avena fatua L. (Poaceae). AB - Allohexaploid wild oat, Avena fatua L. (Poaceae; 2n = 6x = 42), is one of the world's worst weeds, yet unlike some of the other Avena hexaploids, its genomic structure has been relatively little researched. Consequently, in situ hybridisation was carried out on one accession of A. fatua using an 18S-25S ribosomal DNA (rDNA) sequence and genomic DNA from A. strigosa (AA-genome diploid) and A. clauda (CC-genome diploid) as probes. Comparing these results with those for other hexaploids studied previously: (i) confirmed that the genomic composition of A. fatua was similar to the other hexaploid Avena taxa (i.e., AACCDD), (ii) identified major sites of rDNA on three pairs of A/D-genome chromosomes, in common with other Avena hexaploids, and (iii) revealed eight chromosome pairs carrying intergenomic translocations between the A/D- and C genomes in the accession studied. Based on karyotype structure, the identity of some of these recombinant chromosomes was proposed, and this showed that some of these could be divided into two types, (i) those common to all hexaploid Avena species analysed (3 translocations) and (ii) one translocation in this A. fatua accession not previously observed in reports on other hexaploid Avena species. If this translocation is found to be unique to A. fatua, then this information, combined with more traditional morphological data, will add support to the view that A. fatua is genetically distinct from other hexaploid Avena species and thus should retain its full specific status. PMID- 10382297 TI - Rana/Pol III: a family of SINE-like sequences in the genomes of western Palearctic water frogs. AB - The highly repetitive Rana/Pol III family consists of short, tandemly arrayed sequences, scattered throughout the genomes of Palearctic green water frogs. The repeat unit is about 250 bp in length and is a composite element: it contains a SINE-like retroposon with a tRNA structure, flanked by two short direct repeats, and the occurrence of two internal repeats gives evidence that an additional transposition event may have inserted a segment within the already transposed element. Rana/Pol III family is present in the genomes of Rana lessonae, R. ridibunda, and their hybrid form R. esculenta, as well as in R. shqiperica. R. epeirotica, R. cretensis, and the Italian taxon. These sequences are also present in the Iberian R. perezi, although less abundant, but appear to be lacking in the north African species R. saharica. The distribution of Rana/Pol III in the genomes of Palearctic green frogs is in agreement with the phyletic history based on genetic data. The evolutionary pattern proposed for the genus Rana enables us to suppose that the hybridogenetic mechanism is one of the factors accounting for the possible horizontal transfer of Rana/Pol III elements from the central-north Europe species to R. perezi. PMID- 10382299 TI - A fertile amphiploid between diploid wheat (Triticum tauschii) and crested wheat grass (Agropyron cristatum). AB - Alloploidy, one of the most efficient evolutionary mechanisms in nature, has not been extensively exploited in plant breeding programmes. Many genomic combinations remain to be created by plant breeders, to be used directly as new crops or indirectly to widen the genetic basis of crops. The Triticeae tribe, to which wheat belongs, is among the botanical groups in which this strategy has been successfully explored. However, there remain valuable genomic combinations that have not been obtained at the diploid level. The Agropyron complex (wheat grasses) has recently been the focus of attention for interspecific hybridization, but intergeneric hybrids or amphiploids with wheat have not been reported at the diploid level. Here we report synthesis of a tetraploid amphiploid between Triticum tauschii and Agropyron cristatum by crossing two tetraploid accessions. Using total genome in situ hybridization (GISH) staining on metaphase I pollen mother cells, data on allosyndetic and autosyndetic chromosome pairing have been obtained. These data support the view that the A. cristatum tetraploid parent used in the synthesis of the amphiploid has a segmental alloploidy nature. PMID- 10382300 TI - Chromosome evolution in kangaroos (Marsupialia: Macropodidae): cross species chromosome painting between the tammar wallaby and rock wallaby spp. with the 2n = 22 ancestral macropodid karyotype. AB - Marsupial mammals show extraordinary karyotype stability, with 2n = 14 considered ancestral. However, macropodid marsupials (kangaroos and wallabies) exhibit a considerable variety of karyotypes, with a hypothesised ancestral karyotype of 2n = 22. Speciation and karyotypic diversity in rock wallabies (Petrogale) is exceptional. We used cross species chromosome painting to examine the chromosome evolution between the tammar wallaby (2n = 16) and three 2n = 22 rock wallaby species groups with the putative ancestral karyotype. Hybridization of chromosome paints prepared from flow sorted chromosomes of the tammar wallaby to Petrogale spp., showed that this ancestral karyotype is largely conserved among 2n = 22 rock wallaby species, and confirmed the identity of ancestral chromosomes which fused to produce the bi-armed chromosomes of the 2n = 16 tammar wallaby. These results illustrate the fission-fusion process of karyotype evolution characteristic of the kangaroo group. PMID- 10382301 TI - Long tomato microsatellites are predominantly associated with centromeric regions. AB - Microsatellites as genetic markers are used in many crop plants. Major criteria for their usability as molecular markers include that they are highly polymorphic and evenly spread throughout a genome. In tomato, it has been reported that long arrays of tetranucleotide microsatellites containing the motif GATA are highly clustered around the centromeres of all chromosomes. In this study, we have isolated tomato microsatellites containing long arrays (> 20 repeats) of the dinucleotide motifs GA, GT, AT, as well as GATA, assessed their variability within Lycopersicon esculentum varieties and mapped them onto a genetic map of tomato. The investigated microsatellite markers exhibited between 1 and 5 alleles in a diverse set of L. esculentum lines. Mapping of the microsatellites onto the genetic map of tomato demonstrates that, as previously shown, GATA microsatellites are highly clustered in the regions of the tomato centromeres. Interestingly, the same centromeric location was now found for long dinucleotide microsatellite markers. Because of this uneven distribution, genetic mapping of the entire tomato genome using long dinucleotide microsatellites will be very difficult to achieve and microsatellite markers with shorter arrays of microsatellites could be more suitable for mapping experiments albeit their lower level of polymorphism. Some microsatellite markers described in this study might provide a useful tool to study the molecular structure of tomato centromeric regions and for variety identification. PMID- 10382302 TI - Evidence for evolutionary conservation of a physical linkage between the human BAF60b, a subunit of SWI/SNF complex, and thyroid hormone receptor interacting protein-1 genes on chromosome 17. AB - The ubiquitously expressed rat BAF60b gene, which codes for a subunit of the multiprotein SWI/SNF complex, was recently identified between the pituitary growth hormone (GH-N) and thyroid hormone receptor interacting protein-1 (TRIP-1) genes. In primates, duplication of the GH-N gene has resulted in the addition of four placenta-specific (GH-V, CS-A, CS-B, and CS-L) genes downstream of the GH-N gene. As part of our study of the effect of remote sequences on the transcriptional regulation of the GH/CS gene family, we showed recently that these genes lie 40 kb upstream of the human TRIP-1 gene. We have now investigated the presence of the human BAF60b gene upstream of the TRIP-1 gene for evidence of evolutionary conservation of this arrangement or its disruption by the recent duplication of the nearby GH-N gene in primates. We report that, as in the rat genome, the human BAF60b gene is in the reverse transcriptional direction relative to the TRIP-1 gene, such that their polyadenylation sites are separated by 93 bp which compares with 92 bp in the rat. Reexamination of reported porcine TRIP-1 sequences also revealed the presence of the BAF60b gene separated by 93 bp, supporting an evolutionary conservation of this arrangement. PMID- 10382303 TI - Separating the contributions to 15N transverse relaxation in a fibronectin type III domain. AB - In proteins, dynamic mobility is an important feature of structure, stability, and biomolecular recognition. Uniquely sensitive to motion throughout the milli- to picosecond range, rates of transverse relaxation, R2, are commonly obtained for the characterization of chemical exchange, and the construction of motional models that attempt to separate overall and internal mobility. We have performed an in-depth study of transverse relaxation rates of backbone 15N nuclei in TNfn3(1-90), the third fibronectin type III domain from human tenascin. By combining the results of spin-echo (CPMG) and off-resonance T1 rho experiments, we present R2 rates at effective field strengths of 2 to 40 krad/s, obtaining a full spectrum of 16 independent R2 data points for most residues. Collecting such a large number of replicate measurements provides insight into intrinsic uncertainties. The median standard deviation in R2 for non-exchanging residues is 0.31, indicating that isolated measurements may not be sufficiently accurate for a precise interpretation of motional models. Chemical exchange events on a timescale of 570 microseconds were observed in a cluster of residues at the C terminus. Rates of exchange for five other residues were faster than the sampled range of frequencies and could not be determined. Averaged 'exchange free' transverse relaxation rates, R2(0), were used to calculate the diffusion tensor for rotational motion. Despite a highly asymmetric moment of inertia, the narrow angular dispersion of N-H vectors within the beta sandwich proves insufficient to define deviations from isotropic rotation. Loop residues provide exclusive evidence for axially symmetric diffusion (Dpar/Dper = 1.55). PMID- 10382304 TI - 'Wave-type' structure of a synthetic hexaglycosylated decapeptide: a part of the extracellular domain of human glycophorin A. AB - The three-dimensional structure of a glycopeptide, His-Thr*-Ser*-Thr*-Ser*-Ser* Ser*-Val-Thr-Lys, with 2-acetamido-2-deoxy-alpha-D-galactose (GalNAc) residues linked to six adjacent amino acids from Thr-10 to Ser-15, was studied by NMR spectroscopy and molecular dynamics (MD) simulations. The hexaglycosylated decapeptide is part of the extracellular domain of human glycophorin A and shows an extended structure of the peptide backbone due to O-glycosylation. Furthermore, each GalNAc residue exhibits one and only one NOE contact from the NHAc proton to the backbone amide proton of the amino acid that the sugar is directly bound to. This indicates a strong preference for the orientation of all GalNAc residues towards the N-terminus. NOE build-up curves were used to determine 42 inter-proton distances that, in connection with phi angles of the peptide backbone obtained from 3J-coupling constants, resulted in constraints for a MD simulation in water. The NMR data and the MD simulations show a preference for an extended backbone structure. The GalNAc residues are located alternatingly on opposite sides of the backbone and reduce the flexibility of the peptide backbone. The conformation of the molecule is relatively rigid and shows a 'wave type' 3D structure of the peptide backbone within the glycosylation cluster. This new structural element is also supported by the unusual CD spectrum of the glycopeptide. PMID- 10382305 TI - Internal motion time scales of a small, highly stable and disulfide-rich protein: a 15N, 13C NMR and molecular dynamics study. AB - Motions of the backbone C alpha H alpha and threonine C beta H beta bonds of toxin alpha were investigated using natural abundance 13C NMR and molecular dynamics. Measurement of the 13C longitudinal and transverse relaxation rates employed ACCORDION techniques together with coherence selection by pulsed field gradients and sensitivity enhancement through the use of preservation of equivalent pathway, thus allowing a considerable reduction of the required spectrometer time. 13C R1, R2, 1H-->13C NOE were obtained, as well as the variations of R1 rho (90 degrees) as a function of the rf field strength. These data were compared to those recorded by 1H and 15N NMR on a labelled sample of the toxin [Guenneugues et al. (1997) Biochemistry, 36, 16097-16108]. Both sets of data showed that picosecond to nanosecond time scale motions are well correlated to the secondary structure of the protein. This was further reinforced by the analysis of a 1 ns molecular dynamics simulation in water. Several C alpha H alpha and threonine C beta H beta experimentally exhibit fast motions with a correlation time longer than 500 ps, that cannot be sampled along the simulation. In addition, the backbone exhibits motions on the microsecond to millisecond time scale on more than half of its length. Thus, toxin alpha, a highly stable protein (Tm = 75 degrees C at acidic pH) containing 61 amino acids and 4 disulfides, shows important internal motions on time scales ranging from 0.1-0.5 ps, to 10 100 ps, 1 ns, and about 30 microseconds to 10 ms. PMID- 10382306 TI - Observation and measurement of internucleotide 2JNN coupling constants between 15N nuclei with widely separated chemical shifts. AB - Scalar coupling correlations between hydrogen bonded 15N nuclei in non Watson Crick base pairs is a critical step in the structure determination of unusual nucleic acids. For observing the 2JNN coupling constant between far upfield N2,N6 (amino) nitrogens and far downfield (N1,N3,N7) nitrogens (separated by 150-160 ppm), the HNN-COSY experiment (Dingley and Grzesiek, 1998) is rather insensitive, due to technical difficulties associated with simultaneous excitation of both extremes of the 15N spectrum. These nuclei may be correlated by treating them in a pseudo-heteronuclear manner, using 15N selective pulses. The wide chemical shift separation allows accurate measurement of the 2JNN coupling constant using spin-echo difference methods. Pulse sequences for observation and measurement of 2JNN coupling constants between amino and N7 nuclei are presented and demonstrated on an A-A mismatch segment of the uniformly (15N,13C) labelled DNA sample, d(GGAGGAT)2. PMID- 10382307 TI - Selective and extensive 13C labeling of a membrane protein for solid-state NMR investigations. AB - The selective and extensive 13C labeling of mostly hydrophobic amino acid residues in a 25 kDa membrane protein, the colicin Ia channel domain, is reported. The novel 13C labeling approach takes advantage of the amino acid biosynthetic pathways in bacteria and suppresses the synthesis of the amino acid products of the citric acid cycle. The selectivity and extensiveness of labeling significantly simplify the solid-state NMR spectra, reduce line broadening, and should permit the simultaneous measurement of multiple structural constraints. We show the assignment of most 13C resonances to specific amino acid types based on the characteristic chemical shifts, the 13C labeling pattern, and the amino acid composition of the protein. The assignment is partly confirmed by a 2D homonuclear double-quantum-filter experiment under magic-angle spinning. The high sensitivity and spectral resolution attained with this 13C-labeling protocol, which is termed TEASE for ten-amino acid selective and extensive labeling, are demonstrated. PMID- 10382308 TI - Preparation of encapsulated proteins dissolved in low viscosity fluids. AB - The majority of proteins are too large to be comprehensively examined by solution NMR methods, primarily because they tumble too slowly in solution. One potential approach to making the NMR relaxation properties of large proteins amenable to modern solution NMR techniques is to encapsulate them in a reverse micelle which is dissolved in a low viscosity fluid. Unfortunately, promising low viscosity fluids such as the short chain alkanes, supercritical carbon dioxide, and various halocarbon refrigerants all require the application of significant pressure to be kept liquefied at room temperature. Here we describe the design and use of a simple cost effective NMR tube suitable for the preparation of solutions of proteins encapsulated in reverse micelles dissolved in such fluids. PMID- 10382309 TI - Application of amino acid type-specific 1H- and 14N-labeling in a 2H-, 15N labeled background to a 47 kDa homodimer: potential for NMR structure determination of large proteins. AB - NMR investigations of larger macromolecules (> 20 kDa) are severely hindered by rapid 1H and 13C transverse relaxation. Replacement of non-exchangeable protons with deuterium removes many efficient 1H-1H and 1H-13C relaxation pathways. The main disadvantage of deuteration is that many of the protons which would normally be the source of NOE-based distance restraints are removed. We report the development of a novel labeling strategy which is based on specific protonation and 14N-labeling of the residues phenylalanine, tyrosine, threonine, isoleucine and valine in a fully deuterated, 15N-labeled background. This allows the application of heteronuclear half-filters, 15N-editing and 1H-TOCSY experiments to select for particular magnetization transfer pathways. Results from investigations of a 47 kDa dimeric protein labeled in this way demonstrated that the method provides useful information for the structure determination of large proteins. PMID- 10382310 TI - [13C]-constant-time [15N,1H]-TROSY-HNCA for sequential assignments of large proteins. AB - The greatly improved sensitivity resulting from the use of TROSY during 15N evolution and amide proton acquisition enables the recording of HNCA spectra of large proteins with constant-time 13C alpha evolution. In [13C]-ct-[15N,1H]-TROSY HNCA experiments with a 2H/13C/15N-labeled 110 kDa protein, 7,8-dihydroneopterin aldolase from Staphylococcus aureus, nearly all correlation peaks seen in the [15N,1H]-TROSY-HNCA spectrum were also detected. The improved resolution in the 13C dimension then enabled a significant number of sequential assignments that could not be obtained with [15N,1H]-TROSY-HNCA without [13C]-constant-time period. PMID- 10382311 TI - Sequential assignment and solution secondary structure of doubly labelled ribonuclease Sa. PMID- 10382312 TI - 1H, 13C and 15N NMR sequence-specific resonance assignments for rat apo S100A1(alpha alpha) PMID- 10382313 TI - Sequence-specific 1H, 13C and 15N assignments of a calcium binding protein from Entamoeba histolytica. PMID- 10382314 TI - Resonance assignments of the Tn916 integrase DNA-binding domain and the integrase:DNA complex. PMID- 10382315 TI - Sequence-specific 1H, 13C and 15N assignment and secondary structure of the apo EH2 domain of mouse Eps15. PMID- 10382316 TI - Novel structures in galactoglucomannans of the lichens Cladonia substellata and Cladonia ibitipocae: significance as chemotypes. AB - The galactoglucomannans of two species of the lichen genus Cladonia, C. substellata and C. ibitipocae, were compared. They were homogeneous on gel filtration chromatography and structurally related, having (1-->6)-linked alpha-D mannopyranosyl main-chains, but were substituted in different patterns by alpha- and beta-D-galacto-, beta-D-gluco- and alpha-D-mannopyranosyl groups. The C-1 portions of their 13C-NMR spectra are typical of the lichen species and indicate differences between the two polysaccharides. Partial acetolysis of the galactoglucomannan from C. substellata gave rise to oligosaccharides and three were identified, namely alpha-D-Manp-(1-->3)-alpha beta-D-Galp, alpha-D-Manp-(1- >2)-alpha beta-D-Manp and alpha-D-Manp-(1-->2)-[beta-D-Glcp-(1-->4)]-alpha beta-D Manp, whereas only the latter two were obtained from that of C ibitipocae. Methylation and Smith degradation data confirmed these results. Whereas the mannobiose represents a common structure in lichen heteropolysaccharides, it is the first time that the other oligosaccharides have been isolated from those of lichens. PMID- 10382318 TI - Aromin-A, an Annonaceous acetogenin from Annona cherimola. AB - Chromatographic fractionation of a MeOH extract from the stems of Annona cherimola led to the isolation of a new non-adjacent, bis-THF ring acetogenin (one THF ring being at C-4 and the other at C-16), aromin-A (1) along with the known cytotoxic acetogenin squamocin (2). The structures of these isolates were established by means of mass spectrometry and spectroscopic techniques. PMID- 10382317 TI - The flavonoids of Tanacetum parthenium and T. vulgare and their anti-inflammatory properties. AB - The lipophilic flavonoids in leaf and flower of Tanacetum parthenium and T. vulgaris have been compared. While those of T. parthenium are methyl ethers of the flavonols 6-hydroxykaempferol and quercetagetin, the surface flavonoids of T. vulgare are methyl ethers of the flavones scutellarein and 6-hydroxyluteolin. Apigenin and two flavone glucuronides are surprisingly present in glandular trichomes on the lower epidermis of the ray florets of T. parthenium. The opportunity has been taken to revise the structures of the four 6-hydroxyflavonol methyl ethers of T. parthenium based on NMR measurements. These are now shown to be uniformly 6- rather than 7-O-methylated. Tanetin, previously thought to be a new structure, is now formulated as the known 6-hydroxykaempferol 3,6,4' trimethyl ether. The vacuolar flavonoids of both plants are dominated by the presence of apigenin and luteolin 7-glucuronides; nine other glycosides were present, including the uncommon 6-hydroxyluteolin 7-glucoside in T. vulgare. When the major flavonol and flavone methyl ethers of the two plants were tested pharmacologically, they variously inhibited the major pathways of arachidonate metabolism in leukocytes. There were significant differences in potency, with the tansy 6-hydroxyflavones less active than the feverfew 6-hydroxyflavonols as inhibitors of cyclo-oxygenase and 5-lipoxygenase. PMID- 10382319 TI - [Immunodeficiencies: an overview and various diagnostic difficulties]. PMID- 10382320 TI - [Physiopathogenesis and molecular bases of the primary immunodeficiencies]. PMID- 10382321 TI - [Primary immunodeficiencies and the major histocompatibility system]. PMID- 10382322 TI - [Flow cytometry in the diagnosis of the primary immunodeficiencies]. PMID- 10382323 TI - [Bone marrow transplantation in the primary immunodeficiencies]. PMID- 10382325 TI - [Secondary immunodeficiencies]. PMID- 10382324 TI - [Gene therapy of primary immunodeficiencies. Present and future]. PMID- 10382326 TI - [The Spanish Registry of Primary Immunodeficiencies. REDIP-1998]. PMID- 10382327 TI - [Reactions to genetic testing]. PMID- 10382328 TI - [Consider celiac disease!]. PMID- 10382329 TI - [Right delivery in the right place?]. PMID- 10382330 TI - [Epidemiology of head injuries in Troms]. AB - This retrospective population-based survey describes the epidemiology of head injury in a defined population in Troms. It includes all 247 patients with head injury referred to the University Hospital of Tromso during 1993. Head injury was defined as physical damage to the brain or skull caused by external force. The annual incidence rate of hospital-referred head injury was 229/100,000 population with a male preponderance of 1.7:1.0. Causes were fall in 62%, traffic accident in 21%, and assault in 7% of the cases. The observed incidence rate is low despite the use of wide inclusion criteria, probably due to a decrease in road traffic accidents and fewer referrals. A further decrease in the number of head injuries in our region may be achieved by preventing falls. PMID- 10382331 TI - [Management of minor head injuries in Norwegian hospitals--can the quality be improved?]. AB - Management protocols for minor head injury should include strategies for early detection of intracranial haematomas. This study focuses on the management of minor head injury in 63 Norwegian hospitals. We report considerable inter hospital variation. In most (81%) hospitals, minor head injury patients were treated by general surgeons. Emergency room evaluation included routine radiological evaluation, usually skull radiography, in 18 (29%) hospitals, and assessment according to the Glasgow Coma Scale (GCS) in 27 (43%). GCS was used during in-hospital observation in 32 (51%) hospitals. 33 (52%) discharged selected minor head injury patients without in-hospital observation. We conclude that the quality of care for minor head injury patients in Norwegian hospitals can be improved through extended use of routine early CT and consistent evaluation according to GCS. PMID- 10382332 TI - [An overview of hypertension studies with calcium antagonists]. AB - Calcium antagonists are widely used in the treatment of hypertension. However, few endpoint studies with calcium antagonists have been done to prove reduction in hypertensive complications. Results of the STONE, SYST-EUR and SYST-CHINA studies show that long-acting calcium antagonists are effective compared to placebo, especially in patients with isolated systolic hypertension and diabetes. Ongoing prospective and randomized trials like STOP II, INSIGHT, NORDIL, ALLHAT and ASCOT will clarify whether calcium antagonists are more effective than well proven diuretics and betablockers. ASCOT will test the hypothesis that amlodipine is more efficacious than atenolol in preventing cardiac complications in 18,000 hypertensive patients with high coronary risk including diabetes (among them, 2,000 in Norway). The study is also randomizing the patients in a factorial design to either atorvastatin or placebo, testing the so-called lipid hypothesis. PMID- 10382333 TI - [Ruptured splenic artery aneurysm in pregnancy--a case report]. AB - Ruptured splenic artery aneurysm in pregnancy is a serious condition with high mortality rates for both mother and child. Early diagnosis and surgical intervention without delay is essential to avoid a fatal outcome. The diagnosis should be considered in pregnant women with upper abdominal pain. We describe a patient with ruptured splenic artery aneurysm in pregnancy. Both mother and child survived. PMID- 10382334 TI - [Screening for adult celiac disease]. AB - This article is a review of literature from Medline and other sources, which shows that coeliac disease is far more prevalent than previously considered. The clinical picture is very diverse, making diagnosis difficult in many patients and calling for great clinical awareness. Even patients with no or few symptoms have biochemical signs of malabsorption, e.g. folate, vitamin, and iron deficiency, and many exhibit osteopenia. Patients with untreated coeliac disease carry a significant risk of developing malignancies. Risk groups for screening are family members, patients with coeliac associated disorders, and patients with uncharacteristic symptoms. Screening among apparently healthy subjects has been carried out for epidemiological purposes, but is not recommended outside protocols. Diagnosing coeliac disease is important because lifelong strict dietary treatment is effective in alleviating symptoms and preventing longterm complications. PMID- 10382335 TI - [Dietary treatment of celiac disease]. AB - Life-long gluten-free diet is the established therapy of coeliac disease. Patients suffering from dermatitis herpetiformis benefit from the same treatment. In Norway the gluten-free diet has excluded oats as well as wheat, rye and barley. The basis for this recommendation was a 1972 report indicating that ten out of 23 children consuming oats as part of their gluten-free diet for at least 18 months developed signs of damage to the intestinal mucosa. During the last decades, the clinical picture of coeliac disease has changed as a result of better diagnostic tools. Controlled clinical trials during the last few years indicate that some patients may tolerate small amounts of oats in their gluten free diet. As a consequence, patients may be confused with regard to what dietary regime is recommended in coeliac disease. Compliance with gluten-free diet is important to secure growth and development, the all-round condition, fertility, bone density and a reduced risk of nutrient deficiency and malignancy. Consensus on dietary treatment is essential. A number of controlled trials are under way and the outcome of these studies will in a few years determine whether oats might be included in the standard gluten-free diet. So far oats are not recommended. The physician who makes the diagnosis is responsible for all patients getting adequate dietary counselling and management. Dietary advice given by health personnel must be consistent. PMID- 10382336 TI - [Early discharge from maternity and neonatal care units--limitations and perspectives]. AB - Early discharge from maternity units has become more frequent over the last decade. The driving forces have been historical, ideological, logistic, and financial. Often early discharge has been implemented without adequate changes in follow-up practices, resulting in increased readmission rates to hospitals. Many infants recalled for metabolic screening have not showed up, with the attending risk of mental retardation. The question of length of maternity stays has not been adequately studied. Established practice should therefore be changed cautiously. Mechanisms for quality control must be in place to ensure early detection of unanticipated problems. Early discharge from neonatal intensive care units is happening in several countries, either due to lack of equipment or bed space, or to reduce the cost of care. In wealthier countries technology-dependent infants are increasingly being cared for at home. These practices have also not been well studied, and probably should be. Of particular interest is the question of parents' ability to cope with the challenges of caring for these infants. PMID- 10382337 TI - [A young woman who wanted to take her diabetes seriously]. PMID- 10382338 TI - [Persistent back pain, working situation and disability pension after lumbago surgery]. AB - 373 patients who underwent lumbar disc surgery at our department were submitted to a follow-up after 2-17 years. The aim of our study was to find quality indicators: absence of complications, postoperative status including work capacity, and patient satisfaction Questionnaires were returned by 95% of the patients and showed that 58% had a persistent but usually light back problem. 23% had changed to less strenuous work and 13% had qualified for a permanent disability pension. These two groups had significantly higher rates of recurrences and reoperations and were significantly less satisfied. We found no significant relationship between these two groups and a strenuous occupation, nor did we find any gender disparity. Those who qualified for a permanent disability pension were significantly older than the rest of the patients. Our results compare well with other reports. PMID- 10382339 TI - [A simple therapeutic regime for subacute back pain]. AB - Low back pain seems to be an intrinsic factor in most people's lives. Its cause is still largely unknown. Several structures have been suggested as important pain generators, but proof is still largely lacking. The prognosis is generally regarded as good, but in some patients acute low back pain progresses into a chronic disorder. Some suggest that our way of handling the problem might be partly responsible for the increase in chronic low back pain during the last forty years. Rather than structural derangement, a functional disturbance in the complex reflex system that coordiantes the network of paraspinal muscles could be the background for the impairment. A guarded behaviour and fear can increase the muscular dysfunction and make the situation worse. A simple regime for an effective fear-avoidance and activation program is outlined. The approach includes a clinical examination combined with information for patients about the nature of the problem, provided in a manner designed to reduce fear and give them reason to resume light normal activity. PMID- 10382340 TI - [Prognosis in back pain]. AB - A series of Norwegian controlled studies of treatment for low back pain are reviewed with regard to the prognostic factors identified. The importance of controlled and randomized studies is emphasized. A satisfactory level of prediction can only be reached by combining medical, psychological, and social factors. Important prognostic factors are lateral mobility, generalized and intense pain, fear of pain, long duration of pain and sickness leaves, poor subjective evaluation of prognosis and ability to work, low trust in one's own ability to control health, low level of physical training and activity before the acute event, and high level of other subjective health complaints. These data are in good agreement with results from other countries. New data on treatment emphasize the importance of activation of the patient, reducing the fear and avoiding bedrest and inactivity. PMID- 10382341 TI - [Psychosocial consequences of presymptomatic genetic testing. A retrospective study of testing for Huntington disease]. AB - We studied the psychological impact and psychosocial consequences of direct presymptomatic testing for Huntington's disease in Norway. We interviewed 30 out of a total of 43 persons at risk for Huntington's disease who had been tested one to three years earlier, and had been through the test program, and 19 of their spouses. We also included 16 persons at risk who had decided not to take the test. 22 persons were non-carriers, and seven carriers. One had decided not to know the answer so far. 13 out of 30 answered that the risk of getting Huntington's disease had influenced choices they had made in their lives, but quite a few did not know that they were at risk before they had grown up. Six couples out of 21 had divorced after the test; only three said it happened because of the test result. The main problem for many of the persons who now know they are non-carriers is that siblings already are sick or know they will get the disease. 15 persons (50%) experienced the need for some kind of psychiatric treatment during the pre-test period, during the test procedure, or after the test. Eight persons said they had wanted a closer follow-up after the test; most of them had got a negative answer. In this study most of those at risk had adapted reasonably well to the test results. Only seven persons out of 30 were found to be carriers in our study. We therefore have reason to believe that among the 13 tested persons who declined to be involved in the study, the majority had been identified as carriers. Our findings may lend support to a hypothesis suggesting two kinds of response to being identified as carrier. According to studies of post-traumatic stress disorders, one group adjusts reasonably well. The other group responds by avoiding follow-up contact with professional teams, which suggests more psychosocial pain and distress. PMID- 10382342 TI - [Genetic causes of hearing loss--status and perspectives]. AB - Hearing impairment, defined as > or = 40 dB hearing loss, is the most prevalent sensory handicap, present in 1:750 children and in 4-36% of adults, depending on age. Genetic factors are of major importance in more than 50% of all hearing loss. An important distinction is made between syndromic deafness and isolated deafness depending on the presence or not of associated manifestations from other organs. The knowledge about genetic deafness has increased dramatically in the last few years. As of March 1999, at least 53 loci for isolated deafness of different types of monogenic inheritance have been identified. Suspected genetic heterogeneity has thus been confirmed. At least 15 genes for syndromic deafness have been cloned, leading to increased biological insight in shared developmental pathways in different species and leading to better diagnostic tools applicable to patients. The identification of a particularly frequent mutation in a gap junction gene, GJB2 (connexin 26), may turn out to be of particular diagnostic importance in the aetiological evaluation of childhood deafness even in isolated cases. Application of early screening tests, like otoacoustic emission (OAE), in combination with genetic tests will facilitate early and specific diagnosis of hearing impairment and thereby improve audiological rehabilitation. Syndromic deafness involves all organs, and care for and evaluation of affected individuals should be a multiprofessional task. PMID- 10382343 TI - [Is supplementation of vitamins and minerals dangerous?]. PMID- 10382344 TI - [Self-development groups for medical students]. PMID- 10382345 TI - [A Swedish commission wants to prevent adverse effects of medical humor]. PMID- 10382347 TI - [Job satisfaction of general practitioners]. PMID- 10382346 TI - [Decentralized obstetrics and the fight between general practitioners and midwives]. PMID- 10382348 TI - [The 80th anniversary of the P. V. Mandryka Central Military Clinical Hospital]. PMID- 10382349 TI - [The creation of local computer networks in the medical establishments of the Ministry of Defense of the Russian Federation]. PMID- 10382350 TI - [Dispensary care: long-years' experience and the new possibilities]. PMID- 10382351 TI - [The organizational and methodological bases of medical reconnaissance in extreme situations]. AB - The article deals with the process of medical reconnaissance organisation in various extreme situations, including local wars and armed conflicts. The authors suggest that there is a definite need in special units and mobile sanitary epidemiological groups, trained and equipped for the task. They also recommend that these units should be formed up in the deployable medical set-ups. PMID- 10382352 TI - [The medicolegal aspects of the prevention and diagnosis of drug addiction]. AB - The article focuses on a number of medical and legal problems in the drug addiction and toxicomania elimination in the Armed Forces. It contains some new information on recent documents and publications of the Ministry of Health and the government on the subject. At the other hand, the authors make an overall assessment of the analysis' possibilities in the country and its Armed Forces. PMID- 10382353 TI - [Middle ear trauma]. PMID- 10382354 TI - [The prevention of stroke: the traditions and the outlook]. AB - The article deals with the clinicians' views on the widely used methods of brain circulation disorders prevention. On their own experience and on the literature data the authors make a conclusion, that insult is being caused by a number of etiological and pathogenic factors. They discuss some methods of hemodynamics reserve estimation and formulate new ideas for the disorders' possible development. PMID- 10382355 TI - [Enhancing the efficacy of ophthalmologic surgical care in cataract]. PMID- 10382356 TI - [The role of surgical trauma in the development of infectious complications in chronic calculous cholecystitis]. PMID- 10382357 TI - [The late treatment results in pheochromocytoma]. PMID- 10382358 TI - [The characteristics of surgical cholangiography]. PMID- 10382359 TI - [The treatment procedure in acute Mycoplasma pneumonia]. PMID- 10382360 TI - [The clinical variants of peptic ulcer exacerbations]. PMID- 10382362 TI - [Ultrasonic studies in diseases of the prostate]. PMID- 10382361 TI - [The correction of water-salt metabolism in the intensive therapy of an ischemic stroke]. PMID- 10382363 TI - [Hemostatic changes in acute disorders of the cerebral circulation]. PMID- 10382364 TI - [Methods for the functional prognosis of the visual analyzer for the selection of candidates for flight training]. PMID- 10382365 TI - [The outlook for the use of new physiotherapy technologies]. PMID- 10382366 TI - [Physicians on board the experimental submarine Komsomoletz (on the 10th anniversary of the loss of the crew)]. PMID- 10382367 TI - [The history of the instructional work at the P. V. Mandryka Central Military Clinical Hospital]. PMID- 10382368 TI - [The development of gynecological care at the P. V. Mandryka Central Military Clinical Hospital]. PMID- 10382369 TI - [The medical service of the Navy in World War II]. PMID- 10382370 TI - Technical network for logistics in health (Technet). PMID- 10382371 TI - Marburg fever, Democratic Republic of the Congo. PMID- 10382372 TI - Dracunculiasis surveillance, 1998. PMID- 10382373 TI - Polio, Angola. PMID- 10382375 TI - Progress towards global poliomyelitis eradication, as of May 1999. PMID- 10382374 TI - Future trends in veterinary public health. PMID- 10382376 TI - Hantavirus in the Americas. PMID- 10382378 TI - Linking more men with health care. PMID- 10382379 TI - Team practice centers on patient. PMID- 10382380 TI - When you suspect elder abuse. PMID- 10382381 TI - Analyzing exam questions. PMID- 10382382 TI - Common but curable. Responding to symptoms of testicular cancer. AB - Testicular cancer is one of the most common cancers in men between the ages of 15 and 34. Symptoms of testicular cancer vary but can include scrotal nodules or swelling and a sensation of fullness or heaviness of the scrotum, which may be interpreted as pain. Four treatment options are available for testicular cancer: surgery to remove the cancer, radiation therapy, chemotherapy and bone marrow transplantation. No matter what a patient's treatment course, follow-up studies and office visits are imperative. PMID- 10382383 TI - Benign prostatic hyperplasia. A review of diagnostic and treatment options. AB - Benign prostatic hyperplasia (BPH) is a noncancerous enlargement of the prostate gland caused by histologic hyperplasia that produces an inward transmission of pressure on the urethra and an increased resistance to urine flow. The dominant risk factors for the disease are age and male gender. Weak urine stream and hesitancy are the cardinal obstructive features in BPH. Other signs and symptoms include inability to terminate micturition abruptly, sensation of incomplete emptying and occasionally, urinary retention. Many men with prostate enlargement can be successfully treated with lifestyle modification and medication. But if symptoms persist, with no significant improvement after 6 months of finasteride or 2 to 3 months of an alpha-1 blocker, consider a urology consultation. Several surgical options are available. PMID- 10382384 TI - Hair-raising. The latest news on male-pattern baldness. AB - The initiating event in balding seems to be an abnormal sensitivity to the male sex hormones. In addition, a multifactorial model is emerging in which hormones affect the hair follicle in a way that causes it to be perceived as a foreign body by the immune system, which then mounts an attack. Several new classes of agents have the potential to treat hair loss. More than 40 U.S. and several hundred foreign patents have been issued for hair-loss treatment agents. As is common in dermatology, no single agent works universally against hair loss, so the treatment process is often one of trial and error. PMID- 10382385 TI - Erectile dysfunction. Newer treatment options don't reduce need for education, counseling. PMID- 10382386 TI - Matters of the heart. Men and coronary heart disease. PMID- 10382387 TI - Machismo and mortality. Why are men reluctant to seek health care? PMID- 10382388 TI - Acne in adolescents. Need for treatment is more than skin deep. PMID- 10382389 TI - Managing osteoarthritis pain. PMID- 10382390 TI - Advances in rural health care technology. The Texas example. PMID- 10382391 TI - Diabetes today. An update for patients and clinicians. Centers for Disease Control and Prevention. PMID- 10382392 TI - Collaborating with native healers. PMID- 10382393 TI - Best practice and questions of security. PMID- 10382394 TI - Shared humanity and the psychiatric nurse-patient encounter. AB - The paper is based on a phenomenological study of the nurse-patient encounter, the purpose of which was to uncover meaning and generate understandings of being a psychiatric nurse. The study was informed by the phenomenology of Martin Heidegger (1962) and the philosophical hermeneutics of Hans-Georg Gadamer (1975). Drawing upon this phenomenological study it is my intention to discuss three of the existential elements to emerge from an interpretative analysis of these encounters; 'Being-with' as understanding, 'Being-with' as possibility, and 'Being-with' as 'care-full' concern. The paper also discusses two modes of being with patients; the modes of the 'they' nurse and the 'self' nurse. An underlying theme of shared humanity emerged from the study and had the effect of unifying the other concepts uncovered. PMID- 10382395 TI - Job satisfaction amongst nurses in an interim secure forensic unit in Wales. AB - All grades of nursing staff in a recently established interim secure forensic unit completed the Index of Work Satisfaction (IWS; n = 40). High levels of job satisfaction were achieved on four of the IWS subscales: (i) professional status; (ii) interaction; (iii) doctor-nurse relationship; and (iv) autonomy. Moderate levels of satisfaction were found on two of the subscales, task requirements and administration, while the salary subscale was the major area of dissatisfaction across the group. The high level of overall satisfaction in this area of psychiatric nursing may be regarded as a significant achievement in the development of new clinical services in what is regarded as a stressful area of nursing. Measures of job satisfaction may be useful benchmarks for evaluating future changes and developments in the service and for monitoring and improving the clinical work environment. PMID- 10382396 TI - Children of parents with mental illness. I: An overview from a nursing perspective. AB - This paper explores the psychosocial consequences of parental mental illness for child mental health and the implications for mental health nursing. The literature on risk and vulnerability to psychosocial disorder, resilience, child protection, disorder prevention and epidemiological data are reviewed. Based upon a health promotion approach, a model for mental health nursing advocacy for families of adult consumers is proposed as an effective means of preventing disorder in subsequent generations. PMID- 10382397 TI - Stress and adaptation: preparation for successful retirement. AB - Retirement from work can present a significant adjustment challenge for the older population financially, socially and emotionally. Retirement has been identified as a significantly stressful event. Work provides both a structured activity within a regular time frame and a sense of purpose and daily meaning. Energy is channelled into intellectual, creative, and/or physical tasks that offer a sense of satisfaction when they are completed. The planning for retirement is associated with successful adaptation. A realization of the potential and rewards of retirement will also facilitate this adaptation. PMID- 10382398 TI - Finding the job you want, or, how a girl from Nebraska ended up in Philadelphia. PMID- 10382399 TI - Captain of a sinking ship? PMID- 10382401 TI - The health beliefs, values, and practices of gay adolescents. AB - Barriers to optimal health in the gay adolescent population include a lack of recognition or acceptance by healthcare providers, homophobic attitudes, and an absence of awareness regarding the healthcare needs of this vulnerable population. The literature suggests that gay youths experience such problems as lack of self-esteem, school truancy and dropout, runaway behavior and subsequent homelessness, drug and alcohol abuse, prostitution and sexually transmitted diseases, depression, and suicide. Advanced practice nurses have the opportunity to improve the health of gay youths through recognition, education, outreach, and advocacy. PMID- 10382400 TI - Diabetic neuropathy: pathophysiology and prevention of foot ulcers. AB - Diabetic neuropathy, which affects 60% to 70% of those with diabetes mellitus, is one of the most troubling complications for persons with diabetes, often leading to foot ulcers and potentially to lower limb amputations, both of which are preventable. The physiologic, structural, and functional changes associated with diabetic neuropathy and foot ulcers are discussed. Advanced practice nurses are in a unique position to implement strategies for the prevention of serious and debilitating complications from diabetic neuropathy, including foot assessment, education, and specialist referrals. Research evidence is given to support the use of the Semmes-Weinstein monofilaments to evaluate decreased plantar sensation, a common precursor to ulceration. Ongoing patient and family education can emphasize the importance of preventive self-care measures. Referrals for specialist care and therapeutic footwear can be made by advanced practice nurses. If begun early, these interventions can prevent foot ulcers from diabetic neuropathy, thereby improving the quality of life and reducing healthcare costs for this chronic disease. PMID- 10382402 TI - Aesthetic research: literature. PMID- 10382403 TI - Thinking about the meaning of "research" in nursing. PMID- 10382404 TI - Using research to establish protocols for practice: a statewide study of acute care agencies. AB - The purpose of the study was to examine research utilization practices relative to developing and revising practice protocols in acute care agencies in Delaware. Nurse leaders in 13 acute care agencies identified resource nurses most familiar with the development and revision of agency protocols. Thirty-two resource nurses from 11 agencies, representing critical care, emergency, general medical, general surgical, obstetric, and psychiatric nursing, were interviewed. Interviews were tape-recorded and transcribed. Examples of research-based protocols, defined as those supported by research citations, were obtained. The authors found that the majority of protocols submitted, although referenced, were not research-based. Most institutions used textbooks and standards to support nursing practice protocols. The authors concluded that nurses who are responsible for developing and revising agency protocols were not familiar with the use of research findings to guide the development or revision of protocols and were unsure what constituted the "use of research." PMID- 10382405 TI - Comparison of the oral thermometer versus the tympanic thermometer. AB - After the use of tympanic thermometers replaced the use of oral thermometers at the Veterans Affairs Medical Center in Memphis, the nursing staff initiated a comparison study of the two instruments, monitoring 160 temperature readings. Current studies demonstrate that tympanic thermometers give presumably higher temperature readings than do oral thermometers. The study question asked was: Is there a clinically statistical difference between the measures of the two instruments? A statistically significant difference was found between the readings of the two instruments. Despite published results that infrared thermometers provide readings closer to core temperature than oral thermometers, the oral thermometer registered higher in 69% of the subjects. It may be premature to conclude that the oral thermometer is not as accurate as the tympanic thermometer. Removal of this proven oral system may need to be evaluated, and further comparison studies should be conducted before the tympanic thermometer is unconditionally embraced as the more accurate of the two. PMID- 10382406 TI - Update on federal Medicare rules affecting advanced practice nurses. PMID- 10382407 TI - Looking for rebels. PMID- 10382408 TI - Starting a nursing consultation practice. AB - Because the clinical nurse specialist (CNS) role has been changed or eliminated in many hospital organizations, many CNSs in career transition are considering establishing collaborative or independent nursing consultation practices. Opportunities for consultants exist in diverse practice settings and specialties. Before starting a consultation practice, the CNS should carefully examine goals, identify resources, and begin contacting potential referral sources. He or she must also decide what form of business organization to establish and write a business plan to solidify ideas and prepare for the unexpected. Most CNS consultants rely on personal savings to cover initial business and personal expenses, and many continue working as a CNS until the consultation practice is established. Fees can be set based on community standards, what the market will bear, desired projected income, or a third-party payor's fee schedule. The consultation practice can be marketed by word of mouth, inexpensive advertising techniques such as distributing flyers and business cards, direct mall, and media advertising. In today's healthcare marketplace, opportunities abound for the CNS risk-taker interested in starting a nursing consultation practice. PMID- 10382409 TI - The challenges of a home-based nursing consultation business. AB - The transition from working in a traditional setting to working at home alone can be challenging for new nurse consultants. Home-based consultants can use a variety of strategies to stay focused and connected, such as having a designated work area, limiting distractions, and networking. Nurse consultants can obtain information about business management from community resources, and computer on line services offer a means of contacting other small-business owners. Ongoing business evaluations, which include professional accomplishments as well as an examination of income and expenses, help in planning. Home-based nurse consultants can increase the likelihood of business success by setting objectives, working diligently, and networking with others in the business community. PMID- 10382411 TI - PEGS in children: nursing considerations. AB - The nursing care of children with a percutaneous endoscopic gastrostomy (PEG) means caring for the whole of the child with a PEG. Even in the highly technical and specialized field of PEG, the main themes that characterize pediatric nursing are used as a reference guide (Dall'Oglio, 1996). PMID- 10382410 TI - Defensible documentation using the endoscopy pathway. AB - Complete, defensible documentation in a busy endoscopy unit can be elusive. Quick turnaround time, impatient physicians, and add-on and emergent cases all contribute to chaotic days. The endoscopy pathway provides a guided path for succinct documentation by the healthcare worker. A scoring system is initiated on admission. Assessment parameters clearly define scores for vital signs, level of consciousness, abdominal status, pain, safety, and mobility. Scores can be scanned quickly and compared by the nurse to identify significant changes in each patient. Documentation is timely, communicative, and complete. The endoscopy pathway is illustrated by a case study. It reflects patients' progress throughout their stay in the GI unit from admission to discharge. The implementation process of this innovative chart form is presented. PMID- 10382412 TI - Developing an EndoCaseTrac. AB - Many smaller hospitals are merging with large facilities to form healthcare groups. Standardization is the key to cost effectiveness and consistency within such groups. Not only should standardization be applied to equipment, but also to various aspects of documentation. A multi-facility Endoscopy Nurses Task Force was formed to develop an all-encompassing, easy flowing conscious sedation nursing record. The end product was an EndoCaseTrac that is thorough, complete, and multi purposed, to be used in any situations where intravenous conscious sedation is administered, but with the option of being department-specific. Staff education prior to implementation of the EndoCaseTrac played a very important part in the overall acceptance of this newly developed, very thorough and complete nursing documentation. Frequent staff meetings and discussions after implementation decreased the resistance and insecurity in using the EndoCaseTrac. A performance improvement program was also developed to evaluate the accurate completion by the endoscopy nurse and to identify problems. PMID- 10382413 TI - Botulinum toxin: a case study. AB - A variety of therapeutic options exist for the treatment of achalasia. This case study explores these options and follows an individual's experiences as he undergoes these treatments from a barium swallow to botulinum toxin injection. PMID- 10382414 TI - Flexible endoscope reprocessing. PMID- 10382416 TI - Guidelines for documentation in the gastrointestinal endoscopy setting. Society of Gastroenterology Nurses and Associates, Inc. PMID- 10382415 TI - Microbial bioburden in endoscope reprocessing and an in-use evaluation of the high-level disinfection capabilities of Cidex PA. AB - Most endoscopy clinics use 2% glutaraldehyde as a high-level disinfectant for reprocessing flexible endoscopes. However, even with contact times greater than 30 minutes, survival of organisms has been documented. We compared the high-level disinfection capabilities of glutaraldehyde (45-minute immersion) with a new peracetic acid germicide (20- to 25-minute immersion). Channels, valve housings, and outer sheaths were sampled to quantify bioburden levels after a patient procedure, after manual cleaning, and after disinfection. Total mean bioburden after clinical use was greater than 6 log10 colony-forming units (CFU). Manual cleaning reduced the bioburden by means of 4.7 log10 CFU (gastroscopes) and 6.2 log10 CFU (colonoscopes). High-level disinfection with the new product was achieved in five of six (product stressed by EPA Reuse Test) and 7 of 10 (product stressed by dilution and organic load) successfully disinfected endoscopes, whereas glutaraldehyde achieved it in 4 of 10 (product stressed by dilution and organic load). We conclude that the new peracetic acid product (20- and 25-minute contact time) is at least as effective as glutaraldehyde (45-minute contact time) for reducing the bioburden of vegetative aerobic organisms in endoscopes. PMID- 10382417 TI - Kenalog-10 (triamcinoclone acetonide). PMID- 10382418 TI - The horse & buggy syndrome or was good enough for grandpa good enough for me? PMID- 10382419 TI - Stroke: current concepts of care. AB - Despite the approval of the use of tissue plasminogen activator for the treatment of acute stroke, less than 10% of all patients with stroke are able to reach a medical facility in time to receive the treatment. This article reviews the current concepts of stroke management and emphasizes the importance of public awareness for the signs and symptoms of stroke and the need for immediate attention when a stroke is suspected. PMID- 10382420 TI - The poststroke journey: from agonizing to owning. AB - Stroke is a leading cause of death among older adults. Despite the high incidence of stroke, little research has been done on the poststroke recovery process. The purpose of this article is to present a descriptive or explanatory model for poststroke rehabilitation and recovery that includes six stages. PMID- 10382422 TI - Reflections on a mother's death. PMID- 10382421 TI - Dysphagia among nursing home residents. AB - As part of a larger study that investigated the social, cultural, clinical, and environmental factors that influenced nutritional intake in two proprietary nursing homes, 82 of 100 residents who were not eating well received a bedside dysphagia screening evaluation by a speech pathologist. This article reports on the dysphagia evaluation results and the consequences of swallowing disorders for nursing home residents. Forty-five of the 82 residents (55%) had some degree of dysphagia, ranging from mild to profound, but only 10 of these 45 residents (22%) had been referred for a dysphagia evaluation. Unrecognized and unmanaged dysphagia may lead to dehydration, malnutrition, aspiration pneumonia, and asphyxiation. Assessment and management of dysphagia also are discussed. PMID- 10382423 TI - Indwelling catheter care: dispelling the misconceptions. AB - Long-term indwelling catheters often are incorrectly used and managed. Nurses insert and manage catheters, yet studies have shown that most nurses have limited scientific knowledge in the area of catheters and their care. This article will review the Agency for Health Care Policy and Research guidelines for recommended catheter use and suggest troubleshooting tips for catheter-related problems. The problems of infection, catheter selection and size, urinary leakage, encrustation, catheter blockage, and inadvertent catheter removal will be addressed. Patient and caregiver education tips also will be listed. PMID- 10382424 TI - Vocalizations among cognitively impaired elders: what is your patient trying to tell you? AB - Unpleasant vocalizations are prevalent among cognitively impaired elders and often are considered disruptive and problematic by their families and caregivers. However, another way to perceive such challenging vocalizations is as a communication attempt by these patients to make their needs and feelings known. This article identifies possible meanings underlying these vocal behaviors and suggests strategies to help reduce their occurrence. PMID- 10382425 TI - A team approach to managing urinary incontinence. AB - Urinary incontinence (UI) affects at least 50% of long-term care residents. With most care being delivered by nursing assistants, a need exists to educate and include these team members when implementing a UI management program. This article describes a pilot program instituted at Tacoma Lutheran Home using a team approach. In addition to the benefits the residents received, 27% of the staff thought incontinence was easier to manage after the program was implemented, and 50% said someone in administration listened to their suggestions and took action. The coordinator thought the program's weaknesses were that the staff required consistent supervision to accomplish tasks in a timely manner and it was extremely time-consuming in addition to the other duties of employee health and staff development. PMID- 10382426 TI - Delegation can be miscalculated. PMID- 10382428 TI - The dynamic duo. Mathy Mezey, EdD, RN, FAAN, FGSA and Terry Fulmer, RN, PhD, FAAN, FGSA. PMID- 10382427 TI - Israeli student nurses' attitudes about physical restraints in acute care settings. AB - A total of 147 nursing students from an academic school of nursing in Israel, affiliated with the Nursing Department at Tel Aviv University, were studied to determine their attitudes toward the use of physical restraints, their knowledge of patient restraint protocol, related reasons, alternatives, follow-up, and reporting. Data were analyzed according to the student's year of learning and experience in restraining. Significant results showed that most students displayed negative attitudes toward restraining. Most were quite knowledgeable of patient restraint protocol, but they acted according to accepted practice in the hospital wards. These findings suggest that nursing educators must play an important role in reinforcing knowledge and improving the practicum of nursing students in the clinical area concerning restraints. This goal will be achieved by various educational strategies. PMID- 10382430 TI - Implementing patient education in the home: concepts and teaching strategies. PMID- 10382429 TI - Preventing blood clots with drugs. PMID- 10382431 TI - Herbal foods. PMID- 10382432 TI - OASIS is here--serious issues arise. PMID- 10382433 TI - OASIS resources. PMID- 10382434 TI - Walkabout. PMID- 10382435 TI - Comparing traditional Medicare and managed care Medicare. AB - The current healthcare environment presents multiple challenges and opportunities. Patients need in-home health services and managed care companies require prior approvals for these services. There are many demands on the time of home healthcare nurses and supervisors. The following suggestions are offered with the goal of saving time and energy while getting the task accomplished so that patients receive the care they need: Cultivate an ideal relationship with your insurers' case managers. Be flexible. Communicate pertinent information to whomever is responsible for seeking approvals. Provide high-quality patient care that is reflected in the documentation. Inform patients of the prior authorization process as it relates to their home healthcare services (Harris, Lynch, 1996). Fischer and Hurst (1997, p. 16) state that home care providers should adopt a managed care mentality. Linda Pulliam (1989) shared a list of survival skills during a presentation. One of these is "make friends with this changing world" (p. 1). This is excellent advice for all healthcare nurses as we meet the current challenges and anticipate the new ones in the 21st century. PMID- 10382436 TI - Managing a child's pain and fever. PMID- 10382437 TI - Preventing pressure ulcers in home care patients. AB - A large proportion of home care patients present with pressure ulcers, and many more patients are at risk. Home care nurses have an opportunity to manipulate favorably certain environmental factors that can prevent pressure ulcers from forming and to develop effective treatment plans for ulcers once they occur. This article discusses the current practice for pressure ulcer prevention. Next month, a companion article will explore treatment strategies for these wounds. PMID- 10382438 TI - Choosing an intravenous securement system. AB - A variety of intravenous securement devices now offer alternatives to the use of gauze and tape. Home health nurses must recognize these devices and be able to choose among the available options in making product recommendations to support enhanced patient outcomes specific to infusion therapy. PMID- 10382440 TI - Home care nursing informatic guidelines. AB - Technology presents benefits and special challenges to home care nurses in their attempt to safeguard patients' rights. This article presents a set of home care informatic standards that can assist nurses in managing patient-sensitive information in computer networks and suggests ways to avoid risks while using informatics. PMID- 10382439 TI - Phlebotomy techniques in the home. AB - The discussion covers venipuncture from the first stage of choosing the correct needle and collection tube through documentation. Remember that each situation is different. Adaptations must be made for each individual patient and nurse. The information in this article is a guide, and each nurse will modify these techniques to suit his or her own individual style. Good hunting! PMID- 10382441 TI - It's a war out there. PMID- 10382442 TI - HHNA develops core curriculum. PMID- 10382443 TI - Elusive hunger. PMID- 10382444 TI - Congress's report card on home care progress: Part II. PMID- 10382445 TI - The new ABCs of home care--abandonment, business, and caring. PMID- 10382446 TI - Loriemil Villa Prado. PMID- 10382447 TI - The privileges of membership. A past NSNA president embraces NSNA. PMID- 10382449 TI - Make the Internet your new career partner. PMID- 10382448 TI - Tips on grassroots lobbying. PMID- 10382450 TI - Why genetics competency is a must. The Human Genome Project. PMID- 10382451 TI - A tour of cyberspace. PMID- 10382452 TI - Empowerment through computers. Nursing informatics and HTML. PMID- 10382453 TI - 1999 Helene Fuld Scholar Winning Essay. My experiences at the International Cancer Congress as a 1998 Fuld Fellow. PMID- 10382454 TI - Combining traditional and RN students in a senior capstone experience: the student perspective. AB - BACKGROUND: This study explored the students' perspective related to combining traditional and RN student populations in a senior capstone course. METHOD: In the final semester in a senior baccalaureate program, nursing students (N = 340) completed an investigator-designed 5-item survey. Data were reviewed independently by three investigators for emergent themes. RESULTS: While the course content was applied differently by two student populations, both appreciated a seminar format that allowed increased creativity, autonomy, and flexibility. CONCLUSIONS: Merits of an integrated course were more explicitly stated by traditional students, who viewed the RNs as a source of reality-based learning. Faculty sensitivity to the unique learning needs of each student population is recommended. PMID- 10382455 TI - Low literacy levels in adults: implications for patient education. AB - BACKGROUND: Patients are being discharged with the need for more complex care in the home. Written information frequently is used in health education. Patients and caregivers must be able to read and understand this information. METHOD: A comprehensive literature review of articles on literacy and health education materials was performed. RESULTS: The literature review revealed that low literacy is more prevalent in the United States than is generally recognized. Health education materials often require a reading level higher than the reading level of most patients. There are a number of tools available for assessing reading levels. CONCLUSION: Findings indicate there are implications for nursing practice, education, and research. There is a need for nurses to be aware of the prevalence of low literacy. They need to know how to accurately assess reading ability and develop material that is at the appropriate level for patients and caregivers. There is a need for research to investigate possible relationships between health status and low literacy and to examine the effects of remedial reading programs on health practices. PMID- 10382456 TI - Using focus group methodology in nursing. AB - BACKGROUND: The need to obtain quality data in a cost-efficient manner spawned an exploration of research methods. The value of focus group methodology for multiple roles in nursing became evident. METHOD: This article describes the purpose, process, and application of focus group methodology for nursing continuing education. RESULTS: Ideas for use of focus group methodology in nursing continuing education are suggested. CONCLUSION: Focus group methodology can enhance the work of nurses in continuing education roles. PMID- 10382457 TI - Six rules for computers and other stumbling blocks to obtaining an advanced degree. AB - BACKGROUND: The decision to return for an advanced degree in nursing, after many years outside the academic environment, can be fraught with uncertainty, doubt, and panic. The majority of returning students attempt to juggle career, family, community obligations, and school simultaneously. Nurse educators are in the perfect position to act as well-informed resource people, facilitators, and mentors for these returning students. METHOD: The expression "experience is the best teacher" proved to be true. The first author's (B.C.) return to a master of science program after 18 years out of school provided the opportunity to "learn as you go." This article was written with the desire to help ease the transition and lessen the fear for others. RESULTS: The first author (B.C.) graduates in May 1999 with a MS as a Family Nurse Practitioner, enriched and stronger because of the experience, enthusiastic, and facing the future of the nursing profession with a positive and energetic outlook. CONCLUSION: Provision of support; networking with other returning students; and understanding the basic tool in modern education, the computer, eases the return for an advanced degree. Nurse educators' involvement in educational growth, both individually and as proponents for nurses who are dependent on those educators' expertise, is vital to nursing and its evolution as a profession. PMID- 10382458 TI - How to work with an interpreter. AB - BACKGROUND: A trip to China provided two nursing educators an opportunity to teach Chinese nurses and, in the process, to discover principles for using an interpreter to translate from English into the students' language. CONCLUSION: The authors present 16 tips for use by instructors who teach non-English-speaking learners. The principles apply for all situations of communicating with native speakers of another language using a translator. PMID- 10382459 TI - A case study approach: developing critical thinking skills in novice pediatric nurses. AB - BACKGROUND: The development of critical thinking skills in pediatric nurses is imperative to ensure quality nursing care is delivered. Many new graduates have had limited exposure to pediatric nursing. METHOD: The case study method is an approach to develop competencies with real or hypothetical situations. RESULTS: Case studies provide an interactive teaching-learning strategy that can be applied to novice pediatric nurses in a variety of settings. CONCLUSION: By using case studies, nurse educators responsible for the professional development of new graduates, help expand learners' critical thinking as they assume increased responsibility for pediatric nursing care. PMID- 10382460 TI - Divergent backgrounds, unified goals: continuing education program for multinational nurses in a hospital in the Middle East. AB - This article reports continuing education as one strategy used to resolve the competence issues of multinational nurses working in Saudi Arabia. Four categories of Staff Nurses (I, II, III, and IV), N = 111, recruited from eight countries participated in a study that describes the outcomes of a training session for nurses at one hospital in Dammam. An average of 12 to 13 nurses attended 1-day courses over a 9-day period to address urgent policies and procedures selected from the hospital's guidelines for patient care. Results from pretests and posttests completed by the participants yielded information that echo differences of the nurses in terms of country of origin and possibly educational background, rather than their recruitment status with which the employers determine remuneration and responsibilities. The findings suggest ways to ensure that future nurses derive more benefit, and that recruiters and employers reconsider the criteria used for recruitment and placement. PMID- 10382461 TI - Continuing education outcomes related to pain management practice. AB - BACKGROUND: Nursing practice outcomes of continuing education need to be measured and reported as one indicator of the value of nursing continuing education. This article makes the case that knowledge gain, the traditional measure of continuing education effectiveness, is not necessarily sufficient to assess changes in nursing practice. METHOD: A pretest/posttest design was used to measure nursing practice outcomes of a continuing education program about pain management. A total of 50 attendees returned both the pretests and posttests and 68 attendees returned the follow-up evaluation. RESULTS: Ninety-four percent of the respondents had improved scores on the posttest. Ninety-one percent of the follow up evaluation respondents stated they had an opportunity to use the new information and 98% stated the use of this information has improved patient care. CONCLUSION: This study found that a continuing education program triggered practice changes. The results of this study correlate with previous research that supports the need for practice outcome measurements. PMID- 10382462 TI - Challenging Spanish: ways for nurses to become bilingual. AB - A desire to provide culturally competent care to an increasing population of non English-speakers may lead some nurses to consider learning another language as a way to minimize cross-cultural communication barriers. Nurses wishing to learn a second language may be surprised at the variety of ways to acquire bilingual skills, particularly in Spanish, the most common foreign language encountered in the United States. Reasons to learn Spanish are discussed and ways to do so are reviewed. Learning Spanish or another language can enrich the nurse-patient relationship, enhance nurses' self-esteem, and advance nurses' employment opportunities. Bilingual proficiency in English and Spanish (or another language) should be regarded as an authentic clinical skill supporting nurses' cultural and clinical competencies. PMID- 10382463 TI - Making use of a lesson from the past. PMID- 10382465 TI - The effect of newborn early discharge follow-up program on pediatric urgent care utilization. AB - INTRODUCTION: The effects of an early discharge follow-up program on the subsequent use of urgent care services by newborn infants at Kaiser Permanente in Fontana, Calif, was evaluated to test the hypothesis that participation in the follow-up program would result in a significant decrease in utilization of urgent care services. METHOD: Through records review, the effects of a new early discharge follow-up program on the rate of use of urgent care services was examined. Use of urgent care services by 324 early discharged newborn infants who were born before the new program was initiated was measured and compared with the use of urgent care services by 315 early discharged newborn infants whose parents had attended the program. RESULTS: Of the infants born before the new program was initiated, 58% used urgent care services before the 2- to 3-week newborn checkup. The group that participated in the new program used urgent care at the lower rate of 28%; thus the hypothesis was confirmed. DISCUSSION: Follow-up programs in which qualified nurse practitioners provide care for both the mother and newborn infant are demonstrated to decrease the utilization of urgent care services and better meet the needs of the infant and mother. Innovative and collaborative nurse practitioner-managed programs such as Great Starts are needed to ensure quality care in today's rapidly changing health care environment. PMID- 10382464 TI - Do apnea monitors decrease emotional distress in parents of infants at high risk for cardiopulmonary arrest? AB - INTRODUCTION: The immediate post-partum period is stressful for most parents. The need to use a home apnea monitor may tax parental coping skills even further. Therefore, we conducted a study to assess the psychosocial consequences of apnea monitoring on parental emotional distress and family functioning. METHOD: We studied 104 parents of infants at high risk for cardiopulmonary arrest. Fifty-two parents had infants who used home apnea monitors, and 52 parents were age-matched and gender-matched control subjects. Data were collected during the infant's hospitalization, and then at 2 weeks, 3 months, and 6 months after discharge. At 1 year, parents were interviewed about their attitudes toward using the apnea monitor. RESULTS: Both groups experienced elevated levels of emotional distress, but the group with infants who used the monitors had significant increases in depression and hostility immediately following discharge from the hospital compared with baseline, whereas the non-monitored group had a significant increase in hostility at 6 months. At 1-year follow-up, the majority of the parents reported that they used the monitor every night, felt more secure in using it, and judged it helpful. DISCUSSION: The immediate period following hospital discharge of infants who need to use a home apnea monitor is characterized by significant emotional distress for the parents, which resolves over time. Anticipatory education and counseling of parents is recommended. PMID- 10382466 TI - Maternal perceptions of lead poisoning in children with normal and elevated lead levels. AB - INTRODUCTION: The purpose of this study was to examine mothers' perceptions of the severity and susceptibility of their children to lead poisoning and to determine if a correlation existed between mothers' knowledge of lead poisoning and their children's blood lead levels. It was thought that mothers of children with lead poisoning (lead levels > or = 10 micrograms/dL) would score lower on a test of their perceptions and knowledge of lead poisoning than would mothers of children with normal lead levels (lead levels < or = 9 micrograms/dL). METHOD: A cross-sectional study comparing scores of a questionnaire completed by mothers whose children had elevated blood lead levels and mothers whose children had normal blood lead levels was conducted. RESULTS: No difference was found in the median test score between the 2 groups. For the correct responses on a question by-question comparison, significant difference existed between groups; however, the percentage of correct responses was not always greater for the mothers of children with normal blood lead levels. DISCUSSION: Mothers' perceptions and knowledge of lead poisoning were not associated with their children's blood lead levels. PMID- 10382467 TI - Common pediatric foot dermatoses. AB - The pediatric foot dermatoses known as tinea pedis, shoe dermatitis, and juvenile plantar dermatosis are seen commonly in clinical practice. A knowledge of the clinical presentations and causes of these disorders can help pediatric and family nurse practitioners identify, diagnose, and treat childhood rashes of the feet. The conventional view in pediatrics is that small children rarely have tinea pedis. However, a 1992 study disputes this claim. Any child who has an atypical or persistent foot rash will need a referral or consultation. PMID- 10382469 TI - Febrile seizures--treatment and prevention or not? PMID- 10382468 TI - Managing the hair and skin of African American pediatric patients. AB - In Africa, the ancestral home of most African Americans, hair is viewed as the epitome of beauty. However, when Africans were brought to America as slaves, they were unable to care for their hair and skin adequately and were exposed to the predominant white culture, which valued straight hair and light skin. As a result, many African Americans lost self-esteem because of the characteristics of their hair and skin. In this article we examine the anatomic and physiologic features of African American hair and skin and typical African American hair and skin care practices. Common African American hair and skin disorders and their management are discussed. The goal of this article is to help primary care providers understand the special hair and skin care required for African American children (as well as other dark-skinned patients). With good patient education, understanding one's own hair and skin characteristics can also support positive self-esteem. PMID- 10382470 TI - If it isn't Turner's syndrome? PMID- 10382471 TI - Preventing fetal alcohol syndrome. PMID- 10382472 TI - The development of advancing clinical practice roles in the United Kingdom: reflections following a study tour of the United States. PMID- 10382473 TI - Safer motherhood in South-East Asia. PMID- 10382474 TI - Quality of care in reproductive health programmes: concepts, assessments, barriers and improvements--an overview. AB - At the end of the first decade of the Safe Motherhood Initiative there are still, at a minimum, 1600 women dying every day from complications of pregnancy and childbirth: this is an intolerable human tragedy. The fact that there are almost 100,000 more maternal deaths annually now compared to 10 years ago, 585,000, must present a challenge to every citizen in society. Policy makers, health professionals, social workers, religious leaders, human-rights advocates and the media all have a responsibility to ask themselves: 'What can I do?' All have a role in affecting quality reproductive-health services, which are essential for the reduction of maternal mortality and morbidity, and are an intrinsic human right. The midwife is the obvious catalyst and linch-pin for this effort in the fabric of society. Three papers will address the issues of quality of care in reproductive-health programmes with particular emphasis on safe motherhood. This, the first, article describes the concepts of quality of care in reproductive health programmes, the determinants of quality improvement, assessment tools for service quality, barriers to quality of care and quality improvement together with examples from relevant countries. The second article will address education issues relative to quality of care, and the third will describe the monitoring and evaluation of quality of care with relevant indicators and project results. PMID- 10382475 TI - The experience of pregnancy, labour and birth of Thai women in Australia. AB - OBJECTIVE: To identify the perceptions and experience of pregnancy care, labour and birth of Thai women in Melbourne, Australia. DESIGN: An ethnographic interview and participant observation with women in relation to pregnancy, labour and birth. SETTING: Melbourne Metropolitan Area, Victoria, Australia. PARTICIPANTS: 30 Thai women who are now living in Melbourne. FINDINGS: Thai women saw antenatal care as an important aspect of their pregnancy and sought care as soon as they suspected they were pregnant. They were more concerned about the well-being of their babies than their own health, therefore they attended all antenatal appointments. In general, these women were satisfied with care during labour, but some also had negative experiences with their caregivers and hospital routine. When asked to compare maternity services between Thailand and Australia, most of the women believed that services in Australia were better. However, women who had had good experiences of childbirth in Thailand, tended to have negative feelings about the Australian experience. There was also evidence in this study that most of these Thai women did not receive adequate information about care. IMPLICATIONS FOR PRACTICE: Women's perceptions and experiences of antenatal care, labour and birth deserve attention, if appropriate and sensitive care is to be provided to women in Australia and elsewhere. It is only when women's voices are heard in all aspects of health-care delivery that we may see better and appropriate health services for women in childbirth. PMID- 10382476 TI - Satisfaction with midwife-managed care in different time periods: a randomised controlled trial of 1299 women. AB - OBJECTIVE: To compare women's satisfaction with midwife-managed care with 'shared care' over three different time periods. DESIGN: Randomised controlled trial. SETTING: Glasgow Royal Maternity Hospital, Glasgow, UK. PARTICIPANTS: 1299 women experiencing normal pregnancy (consent rate: 82%). Six hundred and forty-eight women were randomised to midwife-managed care and 651 to 'shared care'. METHODS: Three self-report questionnaires were sent to women's homes. The questionnaires examined: satisfaction with antenatal care at 34-35 weeks' gestation, and satisfaction with intrapartum, hospital- and home-based postnatal care at seven weeks postnatally. The third questionnaire reviewed satisfaction with intrapartum care seven months after delivery. FINDINGS: Women in both groups were satisfied. However, women in the midwife-managed group were more highly satisfied in relation to the dimensions examined: relationships with staff, information transfer, choices and decisions, and social support. The differences between the two groups were evident for all time periods (i.e. antenatal, intrapartum and postnatal periods) and were sustained at seven-month follow-up. This is illustrated in the mean scores for relationships with staff, as measured at 34-35 weeks' gestation (possible range -2; very negative attitudes to 2; very positive attitudes). Women in the midwife-managed group scored a mean of 1.22 compared to 0.74 for the 'shared care' group (mean diff: 0.48; 95% CI: 0.42 to 0.55). While women in both groups were more likely to make positive rather than negative comments in open-ended questions, the midwife-managed group were more likely to make positive comments whereas the 'shared care' group were more likely to make negative comments. CONCLUSION: Midwife-managed care for healthy pregnant women which is integrated into existing services improves satisfaction with antenatal, intrapartum and postnatal care. PMID- 10382477 TI - Promoting successful breast feeding among women with a low income. AB - OBJECTIVE: To identify those factors which promote or discourage successful breast feeding in a sample of women with a low income. DESIGN: Qualitative research using in-depth, semi-structured interviews. SAMPLE: All women with a low income who were identified as having breast fed their latest baby at least once and who had delivered at a district general hospital in the south west of England from 17 September 1996 to 5 February 1997. FINDINGS: Three behavioural areas which determined whether or not women with a low income continued to breast feed were identified: individual and social environmental, baby and midwifery practice factors. In terms of individual and social environmental factors those women who continued to breast feed were more likely to have: positive attitudes; realistic expectations; greater levels of self-esteem; a supportive mother/friend; a partner who was not against breast feeding; and the ability to cope with the perceived temporary social isolation. In terms of baby factors those women who continued to breast feed had babies who were more likely to: have three- or four hourly feeds; be perceived as a contented baby; and have gained weight. And finally, in terms of midwifery practice factors, those women who continued to breast feed were more likely to have: not been separated from their baby; not been given supplementary or complementary feeds; received good advice, especially with regard to positioning the baby at the breast; had greater continuity of midwifery input; had sufficient quality time with a midwife; and had the opportunity to solve problems with a community midwife's help. IMPLICATIONS FOR PRACTICE: Midwifery practice can be improved to promote successful breast feeding among women with a low income by: 1) creating realistic expectations and increasing women's confidence/desire to succeed in breast feeding; 2) providing good quality advice and support to mothers of newborn babies, particularly with regard to positioning the baby at the breast; 3) improving social-support networks available to breast-feeding mothers, perhaps through educating grandmothers (or partners) in breast-feeding matters. PMID- 10382478 TI - Gaining ethical approval: a necessity or an optional extra? PMID- 10382479 TI - Women's experience of pain during childbirth. AB - OBJECTIVE: To describe women's experience of pain during childbirth. DESIGN: A qualitative study using a phenomenological approach. Data were collected by tape recorded interviews. SETTING: An Alternative Birth Care Centre at a university hospital in Sweden in 1995. PARTICIPANTS: Nine women, four primiparous and five multiparous who were two to four days post delivery. KEY FINDINGS: Four themes were identified in the meanings of experience: (1) pain is hard to describe and is contradictory; (2) trust in oneself and one's body; (3) trust in the midwife and husband; and (4) transition to motherhood. The essential structure of the studied phenomenon was described as 'being one's body', which includes a non objectifying view of the body, a presence in the delivery process, and a meaning connected to the transition to motherhood. IMPLICATIONS FOR PRACTICE: The women felt that pain was a natural part of the delivery process, and that the strength and power to cope with it came from within the women. A conclusion is that midwives can help birthing women to find their own ability to cope, and should interfere only if the woman asks or if the natural process is disturbed, e.g. by complications. The experience of pain during childbirth, together with the experience of strength during childbirth, gives meaning to the transition to motherhood. PMID- 10382480 TI - A comparison of women's memories of care during pregnancy, labour and delivery after stillbirth or live birth. AB - OBJECTIVE: To compare women's reports of aspects of their care during pregnancy, labour and delivery following stillbirth and live birth. DESIGN: Data were collected by postal questionnaire in 1994. SETTING: A Swedish nation-wide population-based study of cohorts defined in 1991. PARTICIPANTS: Three hundred and fourteen women with stillbirth (subjects) and 322 women with live birth (controls). MEASUREMENTS AND FINDINGS: Labour and delivery were assessed as physically 'insufferably hard' by 52 (17%) of the subjects and 33 (10%) of the controls. The corresponding figures for emotional strains were 144 (47%) and 21 (7%). Obstetric analgesia was more frequently used during labour for stillbirth. One hundred and thirty-eight (44%) subjects, as compared to 44 (2%) of the controls, left hospital within 24 hours of birth. Almost all the women with stillbirth 296 (95%) stated that it was important to have an explanation of the baby's death. Adverse events related to bromocriptine given to inhibit postpartum lactation, were reported by 60 (22%) of the subjects. KEY CONCLUSIONS: It is possible to ease the distress of labour and delivery for stillbirth. Discussion of the aetiology of the baby's death with the mother should be a priority. The optimal length of stay in hospital after stillbirth remains to be defined. Non pharmacological inhibition of lactation may be presented as an alternative to bromocriptine, breast binding is a concrete 'reality confrontation' for the woman and may aid her in her grieving process. Further studies concerning breast binding vs pharmacological inhibition of lactation and long-term psychological outcome are warranted. PMID- 10382481 TI - Can anxiety in pregnant women be measured using the State-Trait Anxiety Inventory. AB - OBJECTIVE: To explore the stability of the State-Trait Anxiety Inventory (STAI), which was used to explain shifts in women's priorities for intrapartum care. DESIGN: A comparative survey of women's priorities for intrapartum care, using a self-complete questionnaire at two intervals, 34 weeks gestation and 10 days postpartum. The questionnaire incorporated the full 40 item STAI. SETTING: Aberdeen, Scotland. PARTICIPANTS: 217 women presenting at Aberdeen Maternity Hospital at 34 weeks gestation who were 'booked' for delivery within the hospital, and who lived within Aberdeen city. One hundred and thirty-six were expecting their first baby and 81 were expecting their second. MEASUREMENTS: Women's priorities for intrapartum care as ascertained at 34 weeks gestation and 10 days postpartum; shifts in priorities observed during this time period; and factors, such as anxiety (measured by the STAI), which could explain these shifts. FINDINGS: Regardless of parity, women had significantly lower A-Trait scores postnatally than antenatally, when compared over a relatively short time period. Differences between nulliparous and parous women were found where the time lapse, between completion of the scales, was more than 45 days. For nulliparous women significant differences in A-Trait scores were still evident in the 45-56 day interval, but not in the later interval of 57-91 days. Parous women appeared to follow the opposite trend, however the numbers were considerably smaller. CONCLUSION: The findings reported in this paper are derived from a study assessing women's priorities for intrapartum care. Anxiety was not a primary outcome measure in this study, but rather a factor which was measured as a possible explanation for shifts in priorities. However, the findings suggest that the STAI may not be stable around the time of delivery. In particular, the test retest reliability of the STAI A-Trait scale appears to be quite low. IMPLICATIONS: The study reported here raises the need for further research in this area and cautions against the unqualified use of this tool until its performance, specifically in the context of pregnant or recently-delivered women, has been thoroughly assessed. PMID- 10382482 TI - Confusing debriefing and defusing postnatally: the need for clarity of terms, purpose and value. AB - Postnatal debriefing is being recommended in the literature. However, there is little clarity of terminology and there has been no systematic evaluation of this technique. The latter is urgently needed. PMID- 10382483 TI - The RPA. Celebrating 25 years as an advocate for nephrology. PMID- 10382484 TI - Physician practice management: why failure is predictable in the current setting and its impact on nephrology. PMID- 10382485 TI - The impact of workforce issues, accountability for patient outcomes. The specialist's role in the 21st century. Part I. PMID- 10382486 TI - Shifting sand: dialysis funding and modality distribution in Ontario. PMID- 10382487 TI - A Fresenius view of the future of ESRD care. PMID- 10382488 TI - Two new access devices have potential to replace the temporary catheter. PMID- 10382489 TI - 1999 camps for patients with ESRD. PMID- 10382490 TI - Improving employment outcomes: the renal care team's role. PMID- 10382491 TI - The ethical issues of withdrawing from dialysis: a European perspective. PMID- 10382492 TI - Non-heart beating donation: an important contributor to organ procurement. PMID- 10382493 TI - Technology demands quick-change nursing roles. PMID- 10382494 TI - Keep up with changing performance improvement standards. AB - One of the changes to the Joint Commission's Performance Improvement standards noted here is the requirement to collect specific data. PMID- 10382495 TI - Nurses' basic guide to understanding the Medicare PPS. AB - Nurse leaders grappling with changing reimbursement need to know the background and structure of Medicare's Prospective Payment System. The author explains case mix and Resource Utilization Groups, as well as applications for practice. PMID- 10382496 TI - When sexual harassment hits home. AB - Sexual harassment cases illustrate the problems facing employees, including nurses' abusive experiences with both physicians and patients. Learn how to give employees the support they need and lower your liability. PMID- 10382497 TI - Get patient information anytime, anywhere. AB - In a hospital faced with rapidly changing community needs, system integration will unite data from beside monitors and ancillary equipment with the hospital's information system. The technology enables faster, easier, and fewer patient transfers, flexible monitor use, continuous monitoring, and remote access of patient information. PMID- 10382498 TI - Coming to grips with a peer's domestic abuse. PMID- 10382499 TI - Do your nurses delegate effectively? AB - Read how to build a work environment that combines your organization's policies and standards, legal regulations, and nurses' expertise into a safe delegation system with positive impact. PMID- 10382500 TI - Strong leaders strengthen retention. PMID- 10382501 TI - What makes nurses stay? PMID- 10382502 TI - Remember where you came from. PMID- 10382503 TI - Smart safeguards for the ED. AB - Health care facilities can prevent violence in the ED by engaging staff and local police in security planning and education. PMID- 10382504 TI - How to merge caregivers' corporate cultures. AB - Mergers and acquisitions combine sets of employees, as well as corporate entities. These steps show how facilities can proactively address joining corporate cultures for a successful transition. PMID- 10382505 TI - Recognize risk factors to prevent patient falls. AB - Nurse leaders can reduce the rate of falls and injuries resulting from falls by instituting a fall prevention program. This article describes how one hospital developed a successful program. PMID- 10382506 TI - You can have a dedicated pediatric unit. AB - Instead of combining pediatric outpatients with adults to balance patient census, one hospital incorporated outpatient pediatric care in the inpatient unit. The change enables nurses to provide specialized care in a dedicated area and expands the hospital's scope of services. PMID- 10382507 TI - Guide to nursing organizations '99. PMID- 10382508 TI - Linking the physical examination and electrocardiography. AB - This article will help critical care nurse managers raise staff awareness of the crucial link between electrocardiography and the physical examination. PMID- 10382510 TI - Hold the lab in the palm of your hand. AB - Should you consider a point-of-care (POC) testing system? Assess your institution's needs and weigh the advantages of using portable POC blood analyzers. PMID- 10382509 TI - What to do when functional and legal families disagree. AB - In Case 1, a nurse describes a scenario in which a patient's functional and legal families disagree about who has the right to make consent decisions. Case 2 reveals the situation's legal and ethical implications for nurses and how they can facilitate a resolution. PMID- 10382511 TI - Eight steps to help you put consumers first. PMID- 10382512 TI - You're being deposed? Remain calm. PMID- 10382513 TI - Ask AONE's experts ... about counting short-stay census. AB - Three nurse executives offer ways to track short-stay patients for more accurate staff planning. PMID- 10382514 TI - The institutional strengthening of the European Union. PMID- 10382515 TI - Nurse prescribing. PMID- 10382516 TI - Home work. PMID- 10382517 TI - Respectability. PMID- 10382518 TI - Choosing independence. PMID- 10382519 TI - Ethnic minority groups in nursing. PMID- 10382520 TI - The route to successful secondment. PMID- 10382521 TI - Managing learning disabilities. PMID- 10382522 TI - Does your appraisal system measure up? PMID- 10382523 TI - The politics of partnerships. PMID- 10382524 TI - Higher level midwives--who needs them? PMID- 10382525 TI - Cochrane made simple. Smoking cessation programmes implemented during pregnancy. PMID- 10382526 TI - Young, pregnant ... and pleased. PMID- 10382527 TI - Caring for Ann. PMID- 10382529 TI - Antenatal education: past and future agendas. PMID- 10382528 TI - Visiting on the maternity wards. PMID- 10382530 TI - The midwife's role in child protection. Part 1: The legal framework. PMID- 10382531 TI - A beginner's picture of research (IV). Experimental research methods. PMID- 10382532 TI - Serving up nutrition in your antenatal classes. PMID- 10382533 TI - Shoulder dystocia. PMID- 10382534 TI - Pleased to meet you. PMID- 10382535 TI - Which course? Obstetric analgesia and anaesthesia. PMID- 10382536 TI - Baby, it's cold outside. PMID- 10382537 TI - Farnesyltransferase inhibitors: targeting the molecular basis of cancer. PMID- 10382538 TI - Bone marrow transplant for lymphoma: where is the field going? PMID- 10382539 TI - Two MAD tails: what the recent knockouts of Mad1 and Mxi1 tell us about the MYC/MAX/MAD network. AB - Members of the MAD/MXI protein family heterodimerize with MAX and repress transcription by recruiting a chromatin-modifying co-repressor complex to specific DNA target genes. Repression mediated by MAD is thought to antagonize the transcriptional activation and proliferation-promoting functions of MYC-MAX heterodimers. Because they are induced during differentiation, it has been suggested that MAD proteins act to limit cell proliferation during terminal differentiation. There is also controversial evidence that these proteins may function as tumor suppressors. Recently, targeted gene deletions of two members of this gene family, Mad1 and Mxi1, have been carried out in mice. Although these animals display what appear to be quite different phenotypes, further analysis supports the view that both these proteins function in cell-cycle exit during terminal differentiation, and that at least MXI1 can act as a tumor suppressor. PMID- 10382540 TI - Molecular diagnostics of cancer predisposition: hereditary non-polyposis colorectal carcinoma and mismatch repair defects. AB - Hereditary non-polyposis colorectal carcinoma accounts for 5-13% of all colorectal carcinomas and is inherited in a dominant fashion. Two different forms can be distinguished. Type I is restricted to colorectal cancers, whereas type II patients acquire acolorectal, endometrial, gastric, small intestinal and transitional carcinomas of the upper urinary tract. Germline mutations in the human mismatch repair genes (hMSH2, hMSH6, hMLH1, hPMS2) account for the majority of hereditary non-polyposis colorectal carcinoma. As a result of the mismatch repair deficiency, replication errors are not repaired, resulting in a mutator phenotype. Simple repetitive sequences (microsatellites) are especially prone to replication errors and analysis of their stability combined with immunohistochemical analysis of mismatch repair protein expression provides a rapid diagnostic strategy. For patients either (1) fulfilling the Amsterdam criteria for HNPCC, (2) with synchronous or metachronous hereditary non-polyposis colorectal carcinoma-related tumors, (3) with hereditary non-polyposis colorectal carcinoma-related tumors before the age of 45 and/or (4) with right sided CRC and mucinous, solid, or cribriform growth patterns, screening for mismatch repair deficiencies should be performed. The identification of colorectal cancers displaying a mutator phenotype has implications for both treatment and prognosis. PMID- 10382541 TI - XXIII European Symposium on Hormones and Cell Regulation: cell proliferation cascades and protooncogenes. St. Odile near Strasbourg, France, September 25-28, 1998. PMID- 10382542 TI - Gene regulation and cancer. AACR meeting review, the Homestead, Hot Springs, VA, USA, 14-18 October 1998. PMID- 10382543 TI - [Hemodynamic indications for DDD mode pacing therapy]. AB - BACKGROUND AND OBJECTIVE: Because there are limits to the drug treatment or surgical intervention of advanced heart failure, alternative methods are being explored. It was the aim of this study to investigate the extent to which optimal atrioventricular (AV) sequence of dual pacemaker stimulation in patients with advanced dilated cardiomyopathy (DCM) of different aetiology increases their usually much reduced cardiac output. PATIENTS AND METHODS: The study group consisted of 22 patients (five women, 17 men; aged 54-84 years) with heart failure in class I-IV (New York Heart Association classification). Temporary dual electrode stimulations (in the right atrium and ventricle) in the VDD or DDD mode were performed with programmed AV intervals between 80 and 180 ms. Cardiac output (CO), pulmonary capillary and pulmonary arterial pressures were measured via an indwelling catheter. RESULTS: In 17 patients with a 1 degree AV block and/or left bundle branch block in the surface ECG pacing produced a significant increase in CO (from 3.7 +/- 0.75 to 4.6 +/- 0.65 l/min; P < 0.005) and cardiac index (from 2.05 +/- 0.43 to 2.47 +/- 0.37 l/min/m2; P < 0.005). There was no increase in patients with normal PR interval and QRS duration. Mean pulmonary capillary and pulmonary arterial pressures remained unchanged. CONCLUSION: Alteration of AV sequence by pacemaker in selected patients in the late stage of heart failure, particularly if there is also abnormal atrioventricular and/or intraventricular conduction delay, may be an effective complementary method of treatment. PMID- 10382544 TI - [Selective embolization of a bile leak after operative resection of an echinococcal cyst]. AB - HISTORY AND ADMISSION FINDINGS: One year after an Echinococcus granulosa cyst had been resected in a 37-year-old woman she presented for follow-up. She was without symptoms and physical examination was unremarkable. INVESTIGATIONS: Sonography and computed tomography showed renewed growth of the cyst (6.5 cm diameter) in the VIth liver segment. The Echinococcus antibody titre was increased to 1:20 (normal 1:5). DIAGNOSIS, TREATMENT AND COURSE: An echinococcal cyst was again diagnosed and a pericystectomy with intrahepatic drainage performed. A bile leak developed postoperatively through an open bile duct at the operative site. This duct was selectively visualized by endoscopic retrograde choleangiography and its distal part embolized with 1.5 ml Histoacryl: there was no further bile leak. No relevant clinical side effects occurred. CONCLUSION: Selective embolization is a possible alternative to currently employed procedures for stopping bile leaks. PMID- 10382545 TI - [Progressive multifocal leukoencephalopathy as a result of immunosuppressive therapy]. AB - HISTORY AND ADMISSION FINDINGS: A 62-year-old woman developed paresis in her right arm within several weeks. She was being treated with methylprednisolone (4 mg daily) and chlorambucil (2 mg every other day) for systemic lupus erythematodes (SLE), which was now in remission. Neurological examination on admission revealed a right flaccid hemiparesis, predominantly of the right arm. The physical examination was otherwise unremarkable. INVESTIGATIONS: Magnetic resonance imaging (MRI) (T2 weighted) showed hyperintense changes in the subcortical medullary layer of the left precentral gyrus without perifocal oedema or abnormal contrast medium uptake, which argued against progressive cerebral ischaemia or tumour. Blood and cerebrospinal fluid (CSF) showed no abnormalities except leukopenia and a raised antinuclear antibody titre. Progressive paralysis of the right side of the body after 2 years of immunosuppressive treatment, the MRI findings and an essentially normal CSE suggested progressive multifocal leucoencephalopathy (PML), confirmed by polymerase chain reaction (PCR) demonstrating JC-virus DNA in serum and CSF. TREATMENT AND COURSE: As a result of the CNS infection with papovavirus JC, an opportunistic infection of the central nervous system, which is usually fatal, occurred. The cerebral changes spread within a few weeks, despite of the immunosuppressive drugs having been discontinued. The pareses progressed further and a marked personality disorder of organic origin ensued. CONCLUSION: While efficacious immunosuppressive drugs against autoimmune disease are available, their use risks the occurrence of life threatening opportunistic infections. PMID- 10382546 TI - [Interventional bronchoscopic procedures--indications]. PMID- 10382547 TI - [Mutations in TSH receptors: pathogenetic significance and clinical relevance]. PMID- 10382548 TI - [New understanding in the study of infectious diseases. Report of the 25th Ludwig Heilmeyer Symposium of the Society for the Advancement of Internal Medicine]. PMID- 10382549 TI - [Accomplishment of blood alcohol determination in the framework of a social medicine appraisal]. PMID- 10382550 TI - [Atrial fibrillation: a frequent problem in clinical practice]. PMID- 10382551 TI - [Atrial fibrillation: a frequent problem in clinical practice]. PMID- 10382552 TI - [Is a solid intestinal cleansing a hypothesis for the effect of probiotic therapy with "Mutaflor"?]. PMID- 10382553 TI - Selenium. AB - The 4 natural oxidation states of selenium are elemental selenium (0), selenide ( 2), selenite (+4), and selenate (+6). Inorganic selenate and selenite predominate in water whereas organic selenium compounds (selenomethionine, selenocysteine) are the major selenium species in cereal and in vegetables. The principal applications of selenium include the manufacture of ceramics, glass, photoelectric cells, pigments, rectifiers, semiconductors, and steel as well as use in photography, pharmaceutical production, and rubber vulcanizing. High concentrations of selenium in surface and in ground water usually occur in farm areas where irrigation water drains from soils with high selenium content (Kesterson Reservoir, California) or in lakes receiving condenser cooling water from coal-fired electric power plants (Belews Lake, North Carolina). For the general population, the primary pathway of exposure to selenium is food, followed by water and air. Both selenite and selenate possess substantial bioavailability. However, plants preferentially absorb selenates and convert them to organic compounds. Aquatic organisms (e.g., bivalves) can accumulate and magnify selenium in the food chain. Selenium is an essential component of glutathione peroxidase, which is an important enzyme for processes that protect lipids in polyunsaturated membranes from oxidative degradation. Inadequate concentrations of selenium in the Chinese diet account, at least in part, for the illness called Keshan disease. Selenium deficiency occurs in the geographic areas where Balkan nephropathy appears, but there is no direct evidence that selenium deficiency contributes to the development of this chronic, progressive kidney disease. Several lines of scientific inquiry suggest that an increased risk of cancer occurs as a result of low concentrations of selenium in the diet; however, insufficient evidence exists at the present time to recommend the use of selenium supplements for the prevention of cancer. The toxicity of most forms of selenium is low and the toxicity depends on the chemical form of selenium. The acute ingestion of selenious acid is almost invariably fatal, preceded by stupor, hypotension, and respiratory depression. Chronic selenium poisoning has been reported in China where changes in the hair and nails resulted from excessive environmental exposures to selenium. Garlic odor on the breath is an indication of excessive selenium exposure as a result of the expiration of dimethyl selenide. The US National Toxicology Program lists selenium sulfide as an animal carcinogen, but there is no evidence that other selenium compounds are carcinogens. PMID- 10382554 TI - Chromium. AB - Chromium occurs primarily in the trivalent state (III), which is the most stable form, or in the hexavalent state (VI), which is a strong oxidizing agent. Elemental chromium (0) does not occur naturally on earth. Trivalent chromium (III) is an essential trace metal necessary for the formation of glucose tolerance factor and for the metabolism of insulin. Commercial applications of chromium compounds include tanning (III), corrosion inhibition, plating, glassware-cleaning solutions, wood preservatives (VI), manufacture of safety matches, metal finishing (VI), and the production of pigments (III, VI). Hexavalent chromium (VI) contaminated local soil when chromium waste slag was part of the fill material present in residential, public, and industrial areas. In some urban areas, about two-thirds of the chromium in air results from the emission of hexavalent chromium from fossil fuel combustion and steel production. The remaining chromium in air is the trivalent form. The residence time of chromium in air is < 10 days, depending on the particle size. Trivalent compounds generally have low toxicity and the gastrointestinal tract poorly absorbs these compounds. Hexavalent chromium is a skin and mucous membrane irritant and some of these hexavalent compounds are strong corrosive agents. Hexavalent chromium compounds also produce an allergic contact dermatitis characterized by eczema. Sensitivity to trivalent compounds is much less frequent, but some workers may react to high concentrations of these compounds. Hexavalent chromium is recognized by the International Agency for Research on Cancer and by the US Toxicology Program as a pulmonary carcinogen. The increased risk of lung cancer occurs primarily in workers exposed to hexavalent chromium dust during the refining of chromite ore and the production of chromate pigments. Although individual studies suggest the possibility of an excess incidence of cancer at sites outside the lung, the results from these studies are inconsistent. PMID- 10382555 TI - Therapeutic review: is ascorbic acid of value in chromium poisoning and chromium dermatitis? AB - INTRODUCTION: Repeated topical exposure to chromium(VI) may cause an allergic contact dermatitis or the formation of chrome ulcers. Systemic toxicity may occur following the ingestion of a chromium(VI) salt, from chromium(VI)-induced skin burns, or from inhalation of chromium(VI) occurring occupationally. Soluble chromium(VI) salts are usually absorbed more easily and cross cell membranes more readily than trivalent chromium salts, and, therefore chromium(VI) is more toxic than chromium(III). In experimental studies, endogenous ascorbic acid in rat lung, liver, and kidney and human plasma, effectively reduces chromium(VI) to chromium(III). The administration of exogenous ascorbic acid has been advocated therefore in the treatment of systemic chromium poisoning and chromium dermatitis to enhance the extracellular reduction of chromium(VI) to the less bioavailable chromium(III). REVIEW: In vitro experiments confirm that the addition of ascorbic acid to plasma containing chromium(VI) leads to a dose-dependent reduction of chromium(VI) to chromium(III). In animal studies, parenteral ascorbic acid 0.5-5 g/kg significantly reduced chromium-induced nephrotoxicity when administered 30 minutes before parenteral sodium dichromate and up to 1 hour after parenteral sodium chromate dosing. Parenteral ascorbic acid 0.5-5 g/kg also reduced mortality when given orally up to 2 hours after oral potassium dichromate dosing. However, the administration of parenteral ascorbic acid more than 2 hours after parenteral chromate in these experimental studies did not protect against renal damage, and parenteral ascorbic acid given 3 hours postparenteral chromate increased toxicity. In addition, there is no confirmed clinical evidence that the administration of ascorbic acid lessens morbidity or mortality in systemic chromium poisoning. A possible reason for the lack of benefit of ascorbic acid when administration is delayed, is that chromium(VI) cellular uptake has occurred prior to ascorbic acid administration. Topical 10% ascorbic acid has been claimed to reduce significantly the healing time of experimentally induced chrome ulcers in guinea pigs. The proposed mechanism is reduction on the skin surface of chromium(VI) to chromium(III). Several case reports suggest that topical ascorbic acid is effective in the management of chromium dermatitis but this has not been confirmed in controlled clinical trials and, moreover, the practical difficulties of frequent application are likely to limit its usefulness. DISCUSSION: Based on experimental studies, substantial amounts of ascorbic acid would need to be administered, preferably parenterally, soon after exposure to prevent systemic toxicity from chromium(VI) in humans. However, as ascorbic acid is a metabolic precursor of oxalate, the administration of ascorbic acid in high dose could lead to acute oxalate nephropathy, particularly in the presence of renal failure. While smaller doses of ascorbic acid (e.g., 10 g intravenously) are not toxic, such doses probably will not reduce the mortality from systemic chromium poisoning. CONCLUSION: There is currently insufficient evidence to advocate the use of ascorbic acid in the management of systemic chromium toxicity. Topical ascorbic acid may reduce dermal hexavalent chromium exposure, but this observation must be confirmed in controlled studies. PMID- 10382556 TI - Cobalt. AB - Cobalt is a relatively rare magnetic element with properties similar to iron and nickel. The two valance states are cobaltous (II) and cobaltic (III) and the former is the most common valance used in the chemical industry. Cobalt occurs in nature primarily as arsenides, oxides, and sulfides. Most of the production of cobalt involves the metallic form used in the formation of cobalt superalloys. The term "hard metal" refers to compounds containing tungsten carbide (80-95%) combined with matrices formed from cobalt (5-20%) and nickel (0-5%). For the general population, the diet is the main source of exposure to cobalt. In the occupational setting, exposure to cobalt alone occurs primarily during the production of cobalt powders. In other industrial exposures (e.g., hard metal, diamond polishing), additional agents (tungsten) modulate the toxicity of cobalt. Cobalt is an essential element necessary for the formation of vitamin B12 (hydroxocobalamin); however, excessive administration of this trace element produces goiter and reduced thyroid activity. In 1966, the syndrome "beer drinker's cardiomyopathy" appeared in Quebec City, Canada, and was characterized by pericardial effusion, elevated hemoglobin concentrations, and congestive heart failure. An interstitial pulmonary fibrosis has been associated with industrial exposure to hard metal dust (tungsten and cobalt), but not to cobalt alone. Exposure to cobalt alone produces an allergic contact dermatitis and occupational asthma. Treatment of cobalt toxicity is primarily supportive. PMID- 10382557 TI - Copper. AB - Copper is an essential trace element, which is an important catalyst for heme synthesis and iron absorption. Following zinc and iron, copper is the third most abundant trace element in the body. Copper is a noble metal, like silver and gold. Useful industrial properties include high thermal and electrical conductivity, low corrosion, alloying ability, and malleability. Most of the metallic copper appears in electrical applications. Copper is a constituent of intrauterine contraceptive devices and the release of copper is necessary for their contraceptive effects. The average daily intake of copper in the US is about 1 mg Cu with the primary source being the diet. The bioavailability of copper from the diet is about 65-70% depending on a variety of factors including chemical form, interaction with other metals, and dietary components. The biological half-life of copper from the diet is 13-33 days with bilary excretion being the major route of elimination. Copper sulfate is a gastric irritant that produces erosion of the lining of the gastrointestinal tract. Chronic copper toxicity is rare and primarily affects the liver. Wilson's disease and Indian childhood cirrhosis are examples of severe chronic liver disease that results from the genetic predisposition to the hepatic accumulation of copper. The serum copper concentration ranges up to approximately 1.5 mg/L in healthy persons. Gastrointestinal symptoms occur at whole blood concentrations near 3 mg Cu/L. Chelating agents (CaNa2EDTA, BAL) are recommended in severe poisoning, but there are little pharmacokinetic data to evaluate the effectiveness of these agents. PMID- 10382558 TI - Molybdenum. AB - Molybdenum does not exist naturally in the pure metallic form and of the 5 oxidation states (2-6) the predominant species are Mo(IV) and Mo(VI). Molybdenum rapidly polymerizes to a wide variety of complex polymolybdate compounds in solution. The vast majority of molybdenum is used in metallurgical applications (stainless steel, cast-iron alloys). Ammonium tetrathiomolybdate is an experimental chelating agent for Wilson's disease. For the general population, the diet is the most important source of molybdenum and concentrations in water and air usually are negligible. The average daily dietary intake is about 0.1-0.5 mg m.o. Molybdenum is an essential element with relatively low toxicity. Enzymes containing molybdenum catalyze basic metabolic reactions in the carbon, sulfur, and nitrogen cycles. Elimination of molybdenum occurs via the kidney and usually is complete within several weeks. Molybdenosis (teart) is a form of molybdenum toxicity that produces a disease in ruminants similar to copper-deficiency. Little data are available on the human toxicity of molybdenum. A gout-like syndrome and pneumoconiosis have been associated with excessive concentrations of molybdenum, but the inadequate design of the studies prevents an adequate determination of the etiology of these effects. PMID- 10382559 TI - Nickel. AB - Nickel is an essential element for at least several animal species. These animal studies associate nickel deprivation with depressed growth, reduced reproductive rates, and alterations of serum lipids and glucose. Although there is substantial evidence of an essential status for nickel in animals, a deficiency state in humans has not been clearly defined. Nickel is a silver-white metal with siderophilic properties that facilitate the formation of nickel-iron alloys. In contrast to the soluble nickel salts (chloride, nitrate, sulfate), metallic nickel, nickel sulfides, and nickel oxides are poorly water-soluble. Nickel carbonyl is a volatile liquid at room temperature that decomposes rapidly into carbon monoxide and nickel. Drinking water and food are the main sources of exposure for the general population with the average American diet containing about 300 micrograms Ni/d. Nickel is highly mobile in soil, particularly in acid soils. There is little evidence that nickel compounds accumulate in the food chain. Nickel is not a cumulative toxin in animals or in humans. Almost all cases of acute nickel toxicity result from exposure to nickel carbonyl. The initial effects involve irritation of the respiratory tract and nonspecific symptoms. Patients with severe poisoning develop intense pulmonary and gastrointestinal toxicity. Diffuse interstitial pneumonitis and cerebral edema are the main cause of death. Sodium diethyldithiocarbamate is an investigational drug used to chelate nickel following exposure to nickel carbonyl. Nickel is a common sensitizing agent with a high prevalence of allergic contact dermatitis. Nickel and nickel compounds are well-recognized carcinogens. However, the identity of the nickel compound or compounds, which cause the increased risk of cancer, remains unclear. Currently, there are little epidemiological data to indicate that exposure to metallic nickel increases the risk of cancer, or that exposure to the carcinogenic forms of nickel causes cancer outside the lung and the nasal cavity. PMID- 10382560 TI - Therapeutic review: do diethyldithiocarbamate and disulfiram have a role in acute nickel carbonyl poisoning? AB - INTRODUCTION: Sodium diethyldithiocarbamate and disulfiram have been proposed as effective nickel chelators. This paper examines the value of these compounds in the treatment of acute nickel carbonyl poisoning by reviewing published experimental and clinical data. REVIEW: In 2 studies, parenteral administration of diethyldithiocarbamate 50-100 mg/kg to rats immediately following nickel carbonyl exposure ensured the survival of all animals: Mortality fell from 73% to 8% when diethyldithiocarbamate was administered at 10 minutes in a third study. In the same study, there was no protection when diethyldithiocarbamate was administered at 6 hours, and the mortality was greater, though not significantly different, when diethyldithiocarbamate was administered at 24 hours. In another study in mice, total protection was afforded by diethyldithiocarbamate given at 8 hours but this protection was limited when diethyldithiocarbamate was administered at 24 hours, with diethyldithiocarbamate 100 mg/kg apparently being less protective than diethyldithiocarbamate 50 mg/kg. In 3 studies, oral diethyldithiocarbamate administration was less effective than parenteral administration. There are no adequately controlled clinical studies of the use of diethyldithiocarbamate in acute nickel carbonyl poisoning despite claims that this therapy has been effective in the treatment of several hundred such patients. Disulfiram, a metabolite of diethyldithiocarbamate, offered complete protection against nickel carbonyl-induced toxicity when administered in a dose of 1000 mg/kg to rats immediately following nickel carbonyl exposure. In contrast, disulfiram 500 mg/kg offered no protection and disulfiram 1500 mg/kg appeared to enhance mortality, possibly by increasing brain nickel accumulation. CONCLUSION: Animal studies demonstrate that diethyldithiocarbamate is an effective antidote in acute nickel carbonyl poisoning when it is administered parenterally soon after exposure. However, as no adequately controlled clinical studies have been performed, further clinical data are required before diethyldithiocarbamate can be recommended routinely in acute nickel carbonyl poisoning. If diethyldithiocarbamate is to be employed, it should be administered parenterally soon after exposure as delay in administration may increase nickel carbonyl toxicity. There are currently insufficient data to recommend disulfiram as an alternative to diethyldithiocarbamate even when diethyldithiocarbamate is not available. PMID- 10382561 TI - Vanadium. AB - Vanadium is a steel-grey, corrosion-resistant metal, which exists in oxidation states ranging from -1 to +5. Metallic vanadium does not occur in nature, and the most common valence states are +3, +4, and +5. The pentavalent form (VO3-) predominates in extracellular body fluids whereas the quadrivalent form (VO+2) is the most common intracellular form. Because of its hardness and its ability to form alloys, vanadium (i.e., ferrovanadium) is a common component of hard steel alloys used in machines and tools. Although most foods contain low concentrations of vanadium (< 1 ng/g), food is the major source of exposure to vanadium for the general population. High air concentrations of vanadium occur in the occupation setting during boiler-cleaning operations as a result of the presence of vanadium oxides in the dust. The lungs absorb soluble vanadium compounds (V2O5) well, but the absorption of vanadium salts from the gastrointestinal tract is poor. The excretion of vanadium by the kidneys is rapid with a biological half-life of 20 40 hours in the urine. Vanadium is probably an essential trace element, but a vanadium-deficiency disease has not been identified in humans. The estimated daily intake of the US population ranges from 10-60 micrograms V. Vanadyl sulfate is a common supplement used to enhance weight training in athletes at doses up to 60 mg/d. In vitro and animal studies indicate that vanadate and other vanadium compounds increase glucose transport activity and improve glucose metabolism. In general, the toxicity of vanadium compounds is low. Pentavalent compounds are the most toxic and the toxicity of vanadium compounds usually increases as the valence increases. Most of the toxic effects of vanadium compounds result from local irritation of the eyes and upper respiratory tract rather than systemic toxicity. The only clearly documented effect of exposure to vanadium dust is upper respiratory tract irritation characterized by rhinitis, wheezing, nasal hemorrhage, conjunctivitis, cough, sore throat, and chest pain. Case studies have described the onset of asthma after heavy exposure to vanadium compounds, but clinical studies to date have not detected an increased prevalence of asthma in workers exposed to vanadium. PMID- 10382562 TI - Zinc. AB - The use of zinc in metal alloys and medicinal lotions dates back before the time of Christ. Currently, most of the commercial production of zinc involves the galvanizing of iron and the manufacture of brass. Some studies support the use of zinc gluconate lozenges to treat the common cold, but there are insufficient data at this time to recommend the routine use of these lozenges. Zinc is an essential co-factor in a variety of cellular processes including DNA synthesis, behavioral responses, reproduction, bone formation, growth, and wound healing. Zinc is a relatively common metal with an average concentration of 50 mg/kg soil and a range of 10-300 mg/kg soil. Meat, seafood, dairy products, nuts, legumes, and whole grains contain relatively high concentrations of zinc. The mobility of zinc in anaerobic environments is poor and therefore severe zinc contamination occurs primarily near points sources of zinc release. The recommended daily allowance for adults is 15 mg zinc. The ingestion of 1-2 g zinc sulfate produces emesis. Zinc compounds can produce irritation and corrosion of the gastrointestinal tract, along with acute renal tubular necrosis and interstitial nephritis. Inhalation of high concentrations of zinc chloride from smoke bombs detonated in closed spaces may cause chemical pneumonitis and adult respiratory distress syndrome. In the occupational setting inhalation of fumes from zinc oxide is the most common cause of metal fume fever (fatigue, chills, fever, myalgias, cough, dyspnea, leukocytosis, thirst, metallic taste, salivation). Zinc compounds are not suspected carcinogens. Treatment of zinc toxicity is supportive. Calcium disodium ethylenediaminetetraacetate (CaNa2EDTA) is the chelator of choice based on case reports that demonstrate normalization of zinc concentrations, but there are few clinical data to confirm the efficacy of this agent. PMID- 10382563 TI - Manganese. AB - Manganese is a very hard, brittle metal, which is used to increase the strength of steel alloys. Absorption from the gastrointestinal tract occurs in the divalent and tetravalent forms. Permanganates, which are strong oxidizing agents, have a +7 valence. The principal organomanganese compound is the anti-knock additive, methylcyclopentadienyl manganese tricarbonyl. Manganese is a ubiquitous constituent of the environment comprising about 0.1% of the earth's crust. For the general population, food is the most important source of manganese with daily intake ranging from 2-9 mg Mn. Combustion of gasoline containing methylcyclopentadienyl manganese tricarbonyl releases submicron particles of Mn3O4 that are potentially respirable. Biomagnification of manganese in the food chain probably does not occur. The lungs and gastrointestinal tract absorb some manganese, but the relative amounts absorbed from each site are not known. Homeostatic mechanisms limit the absorption of manganese from the gastrointestinal tract. Elimination of manganese occurs primarily by excretion into the bile. Animal studies indicate that manganese is an essential co-factor for enzymes, such as hexokinase, superoxide dismutase, and xanthine oxidase. However, no case of manganese deficiency in humans has been identified. Manganism is a central nervous system disease first described in the 1800s following exposure to high concentrations of manganese oxides. Manganese madness was the term used to describe the initial psychiatric syndrome (compulsive behavior, emotional lability, hallucinations). More commonly, these workers developed a Parkinson's-like syndrome. Currently, the risks of exposure to low concentrations of manganese in the industrial and in the environmental settings (e.g., methylcyclopentadienyl manganese tricarbonyl in gasoline) are being evaluated with regards to the development of subclinical neuropsychological changes. The American Conference of Governmental and Industrial Hygienists recently lowered the TLV-TWA for manganese compounds and inorganic manganese compounds to 0.2 mg Mn/m3. PMID- 10382564 TI - Neuropsychological aspects of Lyme disease. PMID- 10382565 TI - Neuropsychological deficits in Lyme disease patients with and without other evidence of central nervous system pathology. AB - A small percentage of Lyme patients develop mild to moderate encephalopathic symptoms months to years after diagnosis and treatment. Their symptoms typically include fatigue, memory loss, sleep disturbance, and depression. However, the etiology of this syndrome remains controversial. It is generally thought that Lyme patients with abnormal cerebral spinal fluid (CSF) have a neurological basis to their illness. To further examine this question, we compared Lyme patients with evidence of abnormal CSF, intrathecal antibody to Borrelia burgdorferi, elevated protein, or a positive polymerase chain reaction for B. burgdorferi DNA (n = 14); Lyme patients with normal CSF (n = 18); and healthy controls (n = 15) on a battery of neuropsychological and personality tests. Although both Lyme groups reported memory problems, only the Lyme group with abnormal CSF had measurable memory deficits. Both Lyme groups had higher depression scores than the normal control group, although depression was not correlated with memory scores. It appears that Lyme patients with abnormal CSF may have a neurological basis to their illness, whereas affective symptoms, common to many chronic disorders, may predispose other Lyme patients to the perception of cognitive dysfunction. PMID- 10382566 TI - Relations among indexes of memory disturbance and depression in patients with Lyme borreliosis. AB - This study examined the relation between complaints of memory disturbance and measures of mood and memory functioning in 55 patients with serological evidence of late-stage Lyme Borreliosis (LB). Patients completed the Self-Ratings of Memory Questionnaire (SRMQ) and the Beck Depression Inventory. Memory functioning was assessed with the California Verbal Learning Test. Depressed patients exhibited more frequent complaints of memory disturbance on the SRMQ, although their pattern of responses did not differ from nondepressed patients. There was a significant correlation between subjective memory ratings and self-reported depression (Spearman rho = -.57, p < .001). No relation was observed between subjective memory ratings and objective memory performance. The results indicate subjective complaints of more severe memory disturbance in patients with LB and depression. Particular attention should be paid to the assessment of depression and subjective symptoms of memory disturbance when administering neuropsychological tests of memory functioning in patients with LB. PMID- 10382567 TI - Psychological states and neuropsychological performances in chronic Lyme disease. AB - The neuropsychiatric sequelae of chronic Lyme disease remains unclear. This study sought to characterize the psychological status of a group of participants who met criteria for post-Lyme syndrome (PLS). These measures were then used to examine the influence of psychological status on neuropsychological performances. Thirty PLS participants completed a structured psychiatric interview, the Positive and Negative Affect Schedule, the Lyme Symptom Checklist, and a battery of neuropsychological tests. As a group, the PLS participants did not appear to have an elevated incidence of psychiatric disorders, and psychiatric history was not useful for understanding neuropsychological performances or symptom reports. The mood of the PLS participants was characterized by lowered levels of positive affect (PA) and typical levels of negative affect. This combination can be distinguished from depression and is consistent with previous findings of affect patterns in individuals with chronic fatigue syndrome. PA was also linked to both total symptom severity and severity of cognitive complaints, but not to duration of illness, neurological manifestations at initial diagnosis, or treatment history. Relative to published normative data, neuropsychological performances were not in the impaired range on any measure. Neither psychological status nor symptom report were useful for understanding any aspect of cognitive functioning. It is concluded that decreased PA is the most useful marker of psychological functioning in PLS. PMID- 10382568 TI - Does process-specific slowing account for cognitive deficits in Lyme disease? AB - Although several studies have suggested that cognitive slowing is a symptom in Lyme disease, it is not clear whether this slowing is general or relates to specific cognitive tasks. This study examined cognitive speed in 25 Lyme disease patients using a mental arithmetic task. These patients showed significant impairments when initiating the cognitive processes involved in counting, but performed as well as healthy participants (n = 23) when the number of counting increments increased. Lyme patients also performed a speeded perceptual-motor matching task as well as healthy participants. Lyme-related initiation speed deficits were significantly correlated with performance on standardized neuropsychological tests, including the Trail Making Test and the Digit Symbol Test, but not with self-reported depression. These results suggest that the cognitive deficits seen on speeded tasks are process specific in the Lyme patient group, and are not the result of generalized slowing. PMID- 10382569 TI - The neuropsychological examination of naming in Lyme borreliosis. AB - Although subjective complaints of word finding and naming deficits are commonly reported by patients with Lyme Borreliosis (LB), the existence of these disturbances has not been thoroughly investigated. Forty-four patients with LB and 43 healthy controls were administered a symptom questionnaire, the Boston Naming Test (BNT), the Controlled Oral Word Association Test (COWAT), and a series of category naming tasks. LB patients had a higher rate of complaints of word-finding disturbance (55% vs. 14%). Lower mean scores were observed on the BNT, but not on the COWAT, nor on category naming tasks. Thirty-six percent of the LB sample exhibited BNT scores in the impaired range. BNT scores in this group were correlated with a measure of memory retrieval, but not with verbal fluency indexes. There was no relation between naming scores and depression. LB patients exhibit impairments in word finding that appear to be secondary to a generalized retrieval deficit. PMID- 10382570 TI - Long-term cognitive effects of Lyme disease in children. AB - Most studies of adults infected with Lyme disease (LD) have found adverse cognitive effects from the disease. In contrast, the only controlled study investigating cognitive effects of LD in a pediatric population found no effects after a 2-year follow-up. However, it was questioned whether the negative effects might take longer than 2 years to emerge. Therefore, this investigation reports a 4-year follow-up of the original sample. Twenty-five children with strictly defined LD were compared with 17 sibling control children. A neuropsychological battery was utilized, including assessment of the cognitive areas of IQ, information processing speed, fine-motor dexterity, novel problem solving and executive functioning, short-term and intermediate memory, and acquisition of new learning. In addition, parents' subjective ratings were obtained on the disease's impact on their child's participation in everyday activities. No between-group differences were found for 17 of the 18 neuropsychological test measures, nor were there differences in parents' subjective ratings. Therefore, in contrast with studies of adults with LD, the results of long-term follow-up of the pediatric population continue to strongly support the finding that children treated appropriately for LD have an excellent prognosis for normal cognitive functioning. PMID- 10382571 TI - Laboratory animal models in periodontology. AB - Animal models are needed to objectively evaluate the pathogenesis of human periodontal diseases and its various treatment modalities. Selection of the appropriate animal model depends on the similarity of the periodontium and the nature of the disease to that of humans. The more commonly used animal models for studying the pathogenesis of periodontal disease, use of implants and guided tissue regeneration have been dogs and nonhuman primates. Periodontal disease in rodents has not been found to be as closely related to the human varieties. Rats and hamsters are best suited for caries and calculus research. Ferrets may be a promising new model for studying periodontal disease and calculus formation. Variables unique to each animal species are manifested by a wide range of clinical and histopathological features. Different species have distinct diets, habits, life spans, tissue structures, host defense mechanisms and genetic traits. This article describes the diversity seen in animal models used to study microbiological, immunological, and clinical features of periodontal disease and its prevention and treatment. PMID- 10382572 TI - Microbiological and clinical effects of an antiseptic dental varnish after mechanical periodontal therapy. AB - The aim of this study was to explore the microbiological and clinical effects of an antiseptic dental varnish when applied to periodontally diseased teeth after mechanical therapy. 20 subjects participated in this placebo controlled, double blind prospective longitudinal study. 2 experimental sites with a pocket probing depth > or =5 mm were chosen in each subject. The control varnish, consisting of ethanol, ethylacetate and polyvinylbutyral, was applied to one of the selected teeth and the test varnish, containing 1% chlorhexidine and 1% thymol in addition, was applied to the other one. Clinical parameters were assessed, and microbiological samples were obtained from the two study sites at baseline (6-10 weeks after completion of conventional periodontal therapy), and 2, 4 and 12 weeks thereafter. The mean PLI at baseline was very low and, therefore, only a minimal potential for a further improvement existed. During the 12-week observation period, the mean PLI increased significantly at sites treated with the placebo varnish, while no similar trend for an increase in PLI was detected in the test sites. The bleeding tendency seemed to remain unaffected by the application of the varnish. On the microbiological level, no relevant differences could be detected between placebo and test sites at baseline, or during the follow-up period. In conclusion, the application of a dental varnish with antimicrobial properties after mechanical periodontal therapy had little effect in subjects with good oral hygiene. PMID- 10382573 TI - Antibiotic susceptibility of putative periodontal pathogens in advanced periodontitis patients. AB - In the present study, the antibiotic susceptibility of most prevalent micro organisms in advanced periodontitis patients was evaluated. In 56 patients, pooled subgingival plaque samples were taken from the deepest site of each quadrant and were cultivated anaerobically. From each patient, the 4 most frequently encountered types of bacterial colonies were subcultured and identified (Rapid ID 32 A). From all bacterial species identified in the 224 subcultures, the 4 most prevalent were used for susceptibility testing to tetracycline, metronidazole and amoxicillin/clavulanate using the E Test. The most prevalent microorganisms were Fusobacterium nucleatum (38/214), Peptostreptococcus micros (33/214), Prevotella oralis (33/214) and Porphyromonas gingivalis (32/214). Regarding antibiotic susceptibility it could be shown that minimal inhibitory concentration (MIC) in all cases was below antibiotic concentrations achievable in gingival crevicular fluid. However, antibiotic resistance was seen in 3 to 29% of the investigated microorganisms. PMID- 10382574 TI - Smoking and crevicular fluid levels of IL-6 and TNF-alpha in periodontal disease. AB - Smoking is a well-documented risk factor for periodontal disease, although the mechanisms of its negative influence are not well understood. In the present study, the influence of smoking on the gingival crevicular fluid (GCF) content of the pro-inflammatory cytokines IL-6 and TNF-alpha was investigated in patients with moderate to severe forms of the disease. The study base consisted of 108 patients including 45 current smokers, 28 former smokers and 35 non-smokers. The median GCF sample levels of IL-6 and TNF-alpha were 5.0 pg/ml and 61.0 pg/ml, respectively, for current smokers, 13.0 pg/ml and 51.0 pg/ml, respectively, for former smokers, and 10.0 pg/ml and 12.0 pg/ml, respectively, for non-smokers. The differences between smoking groups with regard to IL-6 were not significant suggesting that the IL-6 content was not influenced by smoking. In contrast, the TNF-alpha content was significantly increased in current smokers as compared to non-smokers confirming our previous observations. The present results in patients with moderate to severe periodontal disease may indicate different mediator functions of IL-6 and TNF-alpha in response to smoking. PMID- 10382575 TI - The gingival plasma cell infiltrate in HIV-positive patients with periodontitis is disorganized. AB - Patients infected with the human immunodeficiency virus (HIV) are highly susceptible to chronic marginal periodontitis (CMP) and the lesion is generally characterized by abundant plasma cell infiltration. HIV-induced reduction of CD4+ T cells may indirectly affect local production of immunoglobulins (Ig). Gingival biopsies taken from 10 HIV+ and 12 HIV- control patients with CMP were washed, fixed in ethanol and embedded in paraffin. Sections were examined after immunohistochemical staining with monoclonal antibodies against IgA, IgA1-2, IgG, IgG1-4, IgM and IgE. Ig-containing cells were counted in 3 separate connective tissue zones (subjacent to pocket epithelium, central zone and subjacent to oral epithelium). HIV+ patients showed a remarkably increased density of all Ig containing cells in the connective tissue zone subjacent to the oral epithelium (p<0.05) and a lower % of IgG2+ cells in the entire gingival section (p<0.05). In HIV+ patients, the density of IgG-containing cells in the gingiva was strongly correlated with the serum IgG concentration. The altered topical distribution might imply impaired restriction of the inflammatory lesion, additional antigenic challenges by unusual microorganisms in the oral cavity, or be secondary to HIV induced dysregulation of the B-cell system. PMID- 10382576 TI - Extensive expression of TGF-beta1 in chronically-inflamed periodontal tissue. AB - The host immune response in chronic marginal periodontitis (CMP) raised against bacteria colonizing the dentogingival area is modulated by cytokines. This study examines the distribution of the transforming growth factor-beta1 containing (TGF beta1+) cells in formalin-fixed and paraffin-embedded gingival specimens from 11 patients with chronic marginal periodontitis and 7 persons with healthy gingiva. Inflamed periodontal tissue contained a 100-fold more TGF-beta1+ cells than healthy gingiva. Diverse morphological TGF-beta1+ cell types were discerned. Double immuno-enzymatic and -fluorescence staining revealed that TGF-beta1+ cells comprised 21-29% macrophages 2-3% T-cells, 3-9% B-cells, 34-35% neutrophilic granulocytes and 7-10% mast cells. The densities of all TGF-beta1+ cell types in CMP were strongly increased in the connective tissue adjacent to the pocket epithelium, in the lamina propria and adjacent to the oral epithelium. In lesions with extensive inflammation, expression was also marked in pocket epithelium. TGF beta1 is an immunosuppressive cytokine that stimulates wound healing. Upregulation of the cytokine in inflamed gingiva may counterbalance for destructive gingival inflammatory responses that are simultaneously taking place in patients with CMP. PMID- 10382577 TI - Longitudinal study of predictive factors for periodontal disease and tooth loss. AB - Longitudinal assessment of risk factors for periodontal disease is necessary to provide evidence that a putative risk factor or risk indicator is a true risk factor. The purpose of the present study was to explore longitudinally a variety of markers as possible periodontal risk factors in subjects with little or no periodontal disease at baseline. 415 subjects with mild or little periodontal disease were examined: medical and dental history; socioeconomic profile, clinical measurements, microbial samples and radiographic assessment of bone height were performed at baseline, and at a follow-up examination 2 to 5 years later. Mean probing pocket depth (PPD) at baseline was 1.99+/-0.37 mm while mean overall change was 0.1 mm which amounts to an annual rate of 0.04 mm. Overall mean clinical attachment level (1.75+/-0.6 mm) at baseline resulted in mean attachment change of 0.28 mm (0.12 mm annually). Alveolar crestal height (ACH) at baseline (mean 2.05+/-0.85 mm) resulting in a mean net loss of 0.1 mm. Approximately 10% of all sites presented for the second visit with attachment loss exceeding the threshold (4.4% annually), while only 2.2% of all sites exhibited attachment gain (0.88% annually). Older individuals exhibited greater mean bone loss but the least amount of attachment loss. Current smokers exhibited greater disease progression compared to non-smokers. Tooth morbidity (0.17 teeth/patient/year) was associated with greater baseline CAL and ACH loss, and an assortment of systemic conditions. Subjects who harbored Bacteroides forsythus (Bf) at baseline had greater loss in ACH; likewise, these subjects experienced greater proportions of losing sites and twice as much tooth mortality compared to Bf-negative patients. Baseline clinical parameters correlated strongly with the outcome, i.e., subjects with deeper mean pocket depth at baseline exhibited greater increase in pocket depth overtime; while subjects with greater attachment loss at baseline exhibited greater attachment loss between the 1st and 2nd visits. PMID- 10382578 TI - Periodontal repair in dogs: histologic observations of guided tissue regeneration with a prostaglandin E1 analog/methacrylate composite. AB - This report describes observations of healing following guided tissue regeneration (GTR) including surgical implantation of the prostaglandin E1 analog misoprostol with calcium-layered methacrylate particles. Critical size, supra alveolar periodontal defects were surgically created around the 3rd and 4th mandibular premolar teeth in 4 beagle dogs. Wound management included soaking with a 24 microg/ml misoprostol solution and implantation of the misoprostol/methacrylate composite. One jaw quadrant per animal was prepared for GTR using expanded polytetrafluoroethylene membranes. The gingival flaps were coronally advanced and sutured to submerge the teeth. The tissues covering the surgical sites daily received topical misoprostol in an oral adhesive over the 4 week healing interval. Upon euthanasia, tissue blocks were prepared for histometric analysis of regeneration of alveolar bone and cementum, root resorption and ankylosis. The defect area underneath the membrane and the density of methacrylate particles were recorded for the GTR defects. The methacrylate particles appeared encapsulated in a dense connective tissue without signs of an inflammatory reaction, some in contact to newly formed bone. Alveolar bone regeneration height averaged (+/-SD) 1.2+/-1.0 and 1.0+/-0.6 mm for GTR and non GTR defects, respectively. Corresponding values for bone regeneration area were 1.3+/-1.0 and 0.7+/-0.5 mm2. Cementum regeneration was confined to the apical aspect of the defects. Small areas of root resorption and ankylosis were observed for all teeth. Bone regeneration area correlated positively to the defect area and negatively to the density of methacrylate particles in the GTR defects. The histologic observations suggest that the methacrylate composite has marginal potential to promote bone and cementum regeneration under provisions for GTR. PMID- 10382579 TI - The effect of xylitol and chlorhexidine acetate/xylitol chewing gums on plaque accumulation and gingival inflammation. AB - Chewing gums may be suitable vehicles for the delivery of xylitol (X) and chlorhexidine acetate (CHX), both of which can aid oral health. The aim of this study was to determine the clinical effectiveness of chewing gums containing X or a combination of X and CHX in a double-blind, randomised, cross over, 5-day clinical trial, with a 9-day washout period in a group of participants over 40 years old. After professional tooth cleaning, 8 subjects (mean age 51.3+/-10.4 years) used in a random order 2 pieces of ACHX (a liquorice flavoured CHX/X) gum, 2 pieces of BCHX (a chocolate mint flavoured CHX/X), 2 pieces of X (a liquorice flavoured X gum) and 1 piece of ACHX. Gums were chewed 2x daily for 15 min and volunteers refrained from all other oral hygiene procedures. Data were analysed using Friedman nonparametric analysis of variance. Plaque indices for chewing 2 pieces of ACHX gum (0.78+/-0.15) and BCHX gum (0.52+/-0.15) were significantly lower (p<0.0006) than for X gum (1.57+/-0.08). The gingival index was significantly greater (p<0.05) for X containing gum than for the other chewing regimes. The subjects' attitudes to the gums were also assessed by structured questionnaires which showed that all gums were easy to chew, did not adhere to dentures, teeth or restorations and that the subjects preferred to chew 2 pellets rather than 1. PMID- 10382580 TI - Periodontal repair in dogs: effect of rhBMP-2 concentration on regeneration of alveolar bone and periodontal attachment. AB - The objective of this study was to evaluate the effect of recombinant human bone morphogenetic protein-2 (rhBMP-2) concentration on regeneration of alveolar bone and cementum, and on associated root resorption and ankylosis. Contralateral, critical size, supra-alveolar, periodontal defects were surgically produced and immediately implanted with rhBMP-2 in an absorbable collagen sponge (ACS) carrier in 8, young adult, male, beagle dogs. 6 animals received rhBMP-2/ACS (rhBMP-2 at 0.05, 0.10, or 0.20 mg/mL; total construct volume/defect approximately 4.0 mL) in contralateral defects following an incomplete block design. 2 animals received rhBMP-2/ACS (rhBMP-2 at 0 and 0.10 mg/mL) in contralateral defects (controls). The animals were euthanised at 8 weeks post-surgery and block sections of the defects were collected for histologic and histometric analysis. Supra-alveolar periodontal defects receiving rhBMP-2 at 0.05, 0.10, or 0.20 mg/ml exhibited extensive alveolar regeneration comprising 86%, 96%, and 88% of the defect height, respectively. Cementum regeneration encompassed 8%, 6%, and 8% of the defect height, respectively. Root resorption was observed for all rhBMP-2 concentrations. Ankylosis was observed in almost all teeth receiving rhBMP-2. Control defects without rhBMP-2 exhibited limited, if any, evidence of alveolar bone and cementum regeneration, root resorption, or ankylosis. Within the selected rhBMP-2 concentration and observation interval, there appear to be no meaningful differences in regeneration of alveolar bone and cementum. There also appear to be no significant differences in the incidence and extent of root resorption and ankylosis, though there may be a positive correlation with rhBMP-2 concentration. PMID- 10382581 TI - Intra-familial distribution of Fusobacterium nucleatum strains in healthy families with optimal plaque control. AB - Fusobacterium nucleatum, a Gram-negative anaerobic rod associated with periodontal disease, is also found in healthy individuals and is considered part of the indigenous oral microflora. Although intra-familial transmission of periodontal pathogens has been documented, there are no data relating transmission of F. nucleatum. This study investigated the distribution of F. nucleatum strains in 4 strictly healthy families. 32 F. nucleatum strains were isolated from 19 individuals (8 parents and 11 children aged 1-13 years). DNA was extracted and digested with the restriction endonucleases EcoRI, TaqI and HindIII. The digests were separated by electrophoresis through 0.8% agarose gels at 40 V overnight, in TBE buffer containing 1 microg/ml ethidium bromide, and photographed. The DNA was transferred to nylon filters by Southern blotting and hybridized with a digoxigenin labelled E. coli rRNA probe (Kit Dig DNA Labelling mixture - Boehringer). Probed DNA was visualized colorimetrically (CSPD Luminescent Detection Kit Boehringer) and photographed (Amersham). We found that 10/11 children shared identical ribotypes with at least one of their respective parents. Some of the children also harbored a unique additional ribotype. On the basis of indistinguishable restriction endonuclease and ribotype patterns these results support the hypothesis that intra-familial transmission of F. nucleatum is possible. PMID- 10382582 TI - Nutritional regulation of leptin in humans. AB - Although leptin was first discussed as an "adipostat" that regulated food intake in accordance with triglyceride stores, it has become clear that leptin's role is much more complex and that a great many unanswered questions persist. Human studies have not shown that serum leptin concentrations can be changed rapidly by meals. Insulin seems to have a modest but immediate effect in attenuating the morning nadir of serum leptin as well as a greater effect manifested after 4-6 h. These changes in serum leptin, like the decreases that occur with fasting, are not accompanied by corresponding changes in adipose leptin mRNA, suggesting regulation by translational control or changes in the rate of leptin degradation, secretion or clearance. There is convincing evidence that insulin increases leptin synthesis and secretion, probably through an insulin-dependent effect on glucose metabolism. This effect of insulin is possibly mediated by the hexosamine pathway. What adipocytes seem to be communicating to the brain is not how much triglyceride they contain but whether they are currently synthesizing or hydrolysing triglyceride. Confounding many studies is the problem of leptin's diurnal rhythm. Because many studies only measured leptin during its morning nadir or examined the effects of insulin or specific nutrients provided after an overnight fast, important information on regulation may have been lost. PMID- 10382583 TI - Is fasting glucose sufficient to define diabetes? Epidemiological data from 20 European studies. The DECODE-study group. European Diabetes Epidemiology Group. Diabetes Epidemiology: Collaborative analysis of Diagnostic Criteria in Europe. AB - AIMS/HYPOTHESIS: The World Health Organisation Consultation recommended new diagnostic criteria for diabetes mellitus including: lowering of the diagnostic fasting plasma glucose to 7.0 mmol/l and introduction of a new category: impaired fasting glycaemia. The diagnostic 2-h glucose concentrations for diabetes and for impaired glucose tolerance were unchanged. This study identifies fasting plasma glucose concentrations predicting a diabetic 2-h plasma glucose of 11.1 mmol/l or more, analyses the sensitivity and specificity of different screening strategies for diabetes and describes the cardiovascular risk profile in people with impaired fasting glycaemia. METHODS: European population based studies (n = 17) or large, representative samples of employees (n = 3) with both fasting and 2-h post load glucose concentrations following 75-g oral glucose tolerance tests were included (18,918 men and 10,190 women). The Iceland study (8881 men and 9407 women) is presented separately as a 50-g glucose load was used. RESULTS: The fasting plasma glucose predicting a 2-h plasma glucose of 11.1 mmol/l or more with optimal sensitivity and specificity was a) 5.8 mmol/l in women and 6.4 mmol/l in men; b) independent of age; c) increased with obesity. Fasting plasma glucose of 7.0/7.8 mmol/l or more predicted a diabetic 2-h plasma glucose with sensitivities of 49.0/29.8% and specificities of 98.2/99.7%, respectively. CONCLUSION/INTERPRETATION: If fasting glucose is used alone, the 31% of diabetic subjects with a non-diabetic fasting glucose but a diabetic 2-h glucose, will not be diagnosed; impaired fasting glycaemia and impaired glucose tolerance do not identify the same people; the risk profile of people with impaired fasting glycaemia depends on 2-h glucose concentrations. Obesity is the main confounder in the association between fasting and 2-h glucose. PMID- 10382584 TI - Record-high incidence of Type I (insulin-dependent) diabetes mellitus in Finnish children. The Finnish Childhood Type I Diabetes Registry Group. AB - AIMS/HYPOTHESIS: In Finland, the incidence of Type I (insulin-dependent) diabetes mellitus in children aged 14 years or under is the highest in the world and the trend in incidence has been increasing. Our aim was to determine the most recent trends in incidence and the age distribution at diagnosis of Type I diabetes. METHODS: Data on the incidence of Type I diabetes in Finland nationwide were obtained from two sources: for the period 1965 to 1986 from the Central Drug Registry of the Social Insurance Institution and for the period 1987 to 1996 from the prospective childhood Type I diabetes registry. The annual incidence was calculated per 100,000 people. The increase and the trend in incidence were estimated by fitting the linear regression model with the annual incidence data. RESULTS: During 1987 to 1993 the incidence of Type I diabetes seemed to be rather stable at 36 per 100,000 per year. The incidence has continued to increase thereafter and reached 45 per 100,000 per year in 1996. The analysis of the long term trend in incidence between 1965 and 1996 showed an absolute incidence increase of 0.67 per year on average being 3.4 % compared with the incidence in 1965. The increase from 1987 to 1996 was highest in very young children 1-4 years old at diagnosis. CONCLUSION/INTERPRETATION: The high incidence of Type I diabetes in Finnish children has thus far not levelled off but is increasing further. If the trend continues, the predicted incidence in Finland will be approximately 50 per 100,000 per year in the year 2010. PMID- 10382585 TI - Karlsburg Type I diabetes risk study of a general population: frequencies and interactions of the four major Type I diabetes-associated autoantibodies studied in 9419 schoolchildren. AB - AIMS/HYPOTHESIS: The Karlsburg Type I (insulin-dependent) diabetes risk study on schoolchildren aims to evaluate the predictive diagnostic value of diabetes associated autoantibodies in the general population. METHODS: We took capillary serum from 9419 schoolchildren, aged 6-17 years, for testing of autoantibodies (AAbs) to glutamic acid decarboxylase (GADA), protein tyrosine phosphatase (IA2A) and insulin (IAA) by 125I-antigen binding. We also tested for autoantibodies to cytoplasmic islet cell antigens (ICA) immunohistochemically. RESULTS: By testing of 9419 sera for the four AAbs at cut-off at or greater than the 98th centile for the radioassayed AAbs and at or greater than 10 Juvenile Diabetes Foundation (JDF) units for ICA, 8.1% of schoolchildren had at least one AAb. We found that 3.04, 2.97, 2.35, and 0.86% had IAA, GADA, IA2A or ICA, respectively. 7.3% had only one AAb and 0.8% (75) had two or more AAbs, reflecting a risk to develop diabetes. Thus, by primary screening by combined testing of GADA and IA2A, 98.7% (74/75) would be identified. At high AAb levels, cut-off at or greater than the 99.8th centile and at or greater than 40 JDF units for ICA, 0.23% (22/9419) of schoolchildren, similar to the disease prevalence of 0.3%, had two or more AAbs. Ten of 17 children tested had reduced (p < 0.001) first-phase insulin secretion by intravenous glucose tolerance test. Six of 22 subjects developed Type I diabetes within a follow-up of 19 +/- 10 months. CONCLUSION/INTERPRETATION: For children older than 5 years the combined anti-GAD/IA2 test with cut-off at or greater than the 98th centile should be used for primary screening followed by testing for IAA and ICA. Subjects at risk for diabetes have two or more AAbs at or greater than the 98th centile. Subjects at risk for rapid progression to Type I diabetes have two or more AAbs at or greater than the 99.8th centile. PMID- 10382586 TI - Early expression and high prevalence of islet autoantibodies for DR3/4 heterozygous and DR4/4 homozygous offspring of parents with Type I diabetes: the German BABYDIAB study. AB - AIMS/HYPOTHESIS: Islet autoantibodies precede the clinical onset of Type I (insulin-dependent) diabetes mellitus. The cumulative development of such autoantibodies in infants followed from birth and in particular infants with high risk HLA genotypes is poorly defined, but such information is essential to design trials to prevent islet autoimmunity. METHODS: HLA genotypes were determined in offspring of parents with Type I diabetes who were followed from birth for at least 2 years (median follow-up: 3.1 years) and who were characterised for the expression of insulin, GAD65, IA-2 and islet cell autoantibodies at birth, 9 months, 2 and 5 years of age. RESULTS: The HLA genotypes DRB1*03/04(DQB1*57non Asp) and DRB1*04/04(DQB1*57non-Asp) were present in 7.1% and 5.0% of offspring of parents with Type I diabetes. The frequency of both genotypes was increased in offspring who developed islet autoantibodies within the first 2 years of life (27.3% vs 5.5%, odds ratio 6.3 [p = 0.002] and 22.7% vs 4.2%, odds ratio 6.6 [p = 0.003]) and half of all offspring who developed antibodies had these genotypes. Other genotypes were not associated with an increase in risk. By life-table analysis, the cumulative risk of developing islet autoantibodies by the age of 2 years was 20% (95% CI 9.4,30.6) for offspring carrying either the DRB1*03104(DQB1*57non-Asp) or the DRB1*04/04(DQB1*57non-Asp) genotype compared with 2.7% (95% CI 1.2,4.2) for offspring without these genotypes (p < 0.0001). CONCLUSION/INTERPRETATION: These data show that early appearance of islet autoantibodies is remarkably frequent for DR3/4 heterozygous and DR4/4 homozygous offspring and indicate that primary prevention could be considered once available in an offspring cohort selected for these genotypes. PMID- 10382587 TI - Comparison of insulin sensitivity tests across a range of glucose tolerance from normal to diabetes. AB - AIMS/HYPOTHESIS: Adequate comparison of the relative performance of insulin sensitivity tests is not yet available. We compared the discrimination of four insulin sensitivity tests, commonly used in vivo, across a range of glucose tolerance. METHODS: Normal (n = 7), impaired glucose tolerant (n = 8) and Type II (non-insulin-dependent) diabetic subjects (n = 9) had in random order two tests from the following: frequently sampled insulin-modified intravenous glucose tolerance test (FSIVGTT-MinMod); homeostasis model assessment (HOMA) and 2-h continuous infusion of glucose with model assessment (CIGMA) with immunoreactive or specific insulin; short insulin tolerance tests (ITT). The discriminatory power of tests was assessed by the ratio of the within-subject standard deviation to the underlying between-subject standard deviation (discriminant ratio - DR). The degree to which tests measured the same variable was assessed by comparing rank correlation with the maximum expected correlation given the imprecision of the tests. The unbiased lines of equivalence taking into account the precision of tests were constructed. RESULTS: Reciprocal fasting plasma insulin (FPI(-1)), HOMA %S and 2-h CIGMA %S, had similar DRs with ITT being less informative. The FSIVGTT-MinMod analysis was able to assess 13 out of 24 subjects and had a performance similar to ITT. Using specific rather than immunoreactive insulin for HOMA-CIGMA did not improve the DR. Reciprocal fasting plasma insulin FPI(-1), HOMA %S, 2-h CIGMA %S and S(I) FSIVGTT intercorrelated more than 90% of the expected rank correlation given the imprecision of the tests, but ITT gave only limited correlation. CONCLUSION/INTERPRETATION: The HOMA-CIGMA test with immunoreactive insulin provides similar information in distinguishing insulin sensitivity between subjects with normal glucose tolerance, those with impaired glucose tolerance and those with Type II diabetes as does FSIVGTT, whereas ITT is less informative. PMID- 10382588 TI - An uncoupling protein 2 gene variant is associated with a raised body mass index but not Type II diabetes. AB - AIMS/HYPOTHESIS: Linkage between markers close to the uncoupling protein 2 and 3 genes (11q13) and resting metabolic rate and a pre-diabetic phenotype have been found. The syntenic region in mouse has been found to be linked to quantitative traits associated with obesity and diabetes. UCP2 and UCP3 could therefore have an important role in body weight regulation and susceptibility to diabetes. We investigated a recently identified variant of the UCP2 gene in exon 8 as a marker for glucose and weight homeostasis. METHODS: Length variation of the UCP2 exon 8 variant was studied by the polymerase chain reaction and agarose gel electrophoresis. Sequence variants of the UCP3 gene were sought by semi-automated DNA sequencing. RESULTS: In 453 South Indian subjects, we found an association in women between the UCP2 exon variant and body mass index (p = 0.018). These findings were replicated in a separate group of South Indian subjects (n = 143, p < 0.001) irrespective of sex. Although no association was found between the UCP2 exon 8 variant and overt obesity in British subjects, the UCP2 genotype of obese women (n = 83) correlated with fasting serum leptin concentration (p = 0.006) in the presence of extreme obesity. These observations could not be explained by tight linkage disequilibrium with a coding region variant in the region of the UCP3 gene of biological significance. Lastly, no association was found between UCP2 and Type II (non-insulin-dependent) diabetes using either a family based design (85 families) or case control study (normal glucose tolerance n = 335, impaired glucose tolerance n = 42, Type II diabetes n = 76). CONCLUSION/INTERPRETATION: We have described a UCP2 gene exon 8 variant that may affect susceptibility to weight gain by influencing regulation of leptin. PMID- 10382589 TI - NMDA receptor subunits are modified transcriptionally and post-translationally in the brain of streptozotocin-diabetic rats. AB - AIMS/HYPOTHESIS: Moderate disturbances of learning and memory were recognized as a complication of diabetes mellitus in patients. The streptozotocin-diabetic rat, an animal model of insulin-dependent diabetes, shows impairments in spatial memory and in long-term potentiation expression. We have studied the effect of experimental diabetes on expression of post-synaptic glutamate N-Methyl-D Aspartate ionotropic receptors and of other key proteins regulating synaptic transmission at the post-synaptic compartment. METHODS: In situ hybridization and Western blot analysis were used to assess expression and protein concentration of N-Methyl-D-Aspartate receptors and alpha-calcium-calmodulin-dependent kinase II. Receptor subunits alphaCaMKII-dependent phosphorylation was studied in post synaptic densities obtained from the hippocampus and cortex of control, streptozotocin-diabetic and insulin-treated rats. RESULTS: The transcript levels of NR1 and NR2A subunits of N-Methyl-D-Aspartate were unchanged in rats with a diabetic duration of 3 months when compared with age-matched control rats. Accordingly, NR1 and NR2A as well as GluR1, GluR2/3, PSD-95 and alphaCaMKII protein concentrations in post-synaptic densities were the same in both control and diabetic rats, whereas the immunoreactivity for NR2B was reduced by about 40%. In addition, the activity of alphaCaMKII on exogenous substrates, such as syntide-2, and the phosphorylation of NR2A/B subunits of N-Methyl-D-Aspartate receptor was reduced in hippocampal post-synaptic densities of streptozotocin diabetic rats as compared with control rats. Furthermore, we show that insulin intervention for 3 months after diabetic duration partially restored both alphaCaMKII activity and NR2B levels. CONCLUSION/INTERPRETATION: N-Methyl-D Aspartate receptor expression and phosphorylation is possibly involved in behavioural and electrophysiological abnormalities observed in streptozotocin diabetic rats. PMID- 10382590 TI - Down regulation of peroxisome proliferator-activated receptorgamma expression by inflammatory cytokines and its reversal by thiazolidinediones. AB - AIMS/HYPOTHESIS: Previous studies show that inflammatory cytokines play a part in the development of insulin resistance. Thiazolidinediones were developed as insulin-sensitizing drugs and are ligands for the peroxisome proliferator activated receptory (PPARgamma). We hypothesized that the anti-diabetic mechanism of thiazolidinediones depends on the quantity of PPARgamma in the insulin resistant state in which inflammatory cytokines play a part. METHODS: We isolated rat PPARgamma1 and gamma2 cDNAs and examined effects of various cytokines and thiazolidinediones on PPARgamma mRNA expression in rat mature adipocytes. RESULTS: Various inflammatory cytokines, such as tumour necrosis factor-alpha (TNF-alpha), interleukin-1alpha (IL-1alpha), IL-1beta, IL-6 and leukaemia inhibitory factor decreased PPARgamma mRNA expression. In addition, hydrogen peroxide, lysophosphatidylcholine or phorbol 12-myristate 13-acetate also decreased the expression of PPARgamma. The suppression of PPARgamma mRNA expression caused by 10 nmol/l of TNF-alpha was reversed 60% and 55% by treatment with 10(-4) mol/l of troglitazone and 10(-4) mol/l of pioglitazone, respectively. The suppression of glucose transporter 4 mRNA expression caused by TNF-alpha was also reversed by thiazolidinediones. Associated with the change of PPARgamma mRNA expression, troglitazone improved glucose uptake suppressed by TNF-alpha. CONCLUSION/INTERPRETATION: Our study suggests that inflammatory cytokines could be factors that regulate PPARgamma expression for possible modulation of insulin resistance. In addition, we speculate that the regulation of PPARgamma mRNA expression may contribute to the anti-diabetic mechanism of thiazolidinediones. PMID- 10382591 TI - Effect of ageing on beta-cell mass and function in rats malnourished during the perinatal period. AB - AIMS/HYPOTHESIS: In a recently developed rat model, maternal food restriction from day 15 of pregnancy until weaning induced low birth weight and a 70% reduction of beta-cell mass in the offspring at day 21 after birth. Subsequent renutrition from weaning was insufficient to fully restore beta-cell mass in young adult rats. The aim of this study is to investigate the long-term consequences of early malnutrition on beta-cell mass and function. METHODS: Oral glucose tolerance tests were done in 3- and 12-month-old animals and beta-cell mass and apoptosis were determined by morphometrical measurements on pancreatic sections. The specific impact of postnatal malnutrition was studied by comparing control animals (C group) with animals malnourished during their fetal life only (R/C group), and animals malnourished during fetal life and until weaning (R group). RESULTS: In 3-month-old R/C animals beta-cell mass reached 8.0 +/- 1.5 mg with no further increase until 12 months (8.1 +/- 1.5 mg), compared with 9.3 +/- 1.9 mg in control rats. Twelve-month-old R/C animals showed normal plasma insulin responses and borderline glucose tolerance. In R animals, apoptosis reached 1.9 +/- 0.4% of the beta cells at 3 months, compared with 0.7 +/- 0.5% in control rats, and beta-cell mass did not increase between 3 and 12 months (4.7 +/- 0.8 mg at 12 months). In aged control and R animals, apoptosis affected 8% of the beta cells. At 12 months only, R animals showed profound insulinopenia and marked glucose intolerance. CONCLUSION/INTERPRETATION: In conclusion, perinatal malnutrition profoundly impairs the programming of beta-cell development. In animals with decreased beta-cell mass the additional demand placed by ageing on the beta cells entails glucose intolerance since beta-cell mass does not expand and apoptosis is increased. PMID- 10382592 TI - Involvement of Smad proteins in the differentiation of pancreatic AR42J cells induced by activin A. AB - AIMS/HYPOTHESIS: Activin A induces differentiation of amylase-secreting pancreatic AR42J cells into endocrine cells. This study assesses the role of Smad proteins in the actions of activin A in AR42J cells. METHODS: The expression of Smad proteins was determined by northern blotting. Phosphorylation and translocation of Smad2 was measured by transfecting flag-tagged Smad2. Involvement of Smad2 was examined by transfecting cDNA encoding N-terminal and C terminal domains of Smad2. RESULTS: The mRNAs for Smad2 and Smad4 were abundantly expressed whereas the expression of mRNA for Smad1 and Smad3 was very low. Activin A induced serine-phosphorylation and the subsequent accumulation of the Smad2 in nuclei. Transfection of the N-terminal domain of Smad2, which acts as a dominantly negative mutant (Smad2-N), blocked the morphological changes induced by activin A whereas the C-terminal domain of Smad2, which acts as a constitutively active mutant (Smad2-C), reproduced the activin-induced morphological changes. Similarly, Smad2-N blocked apoptosis induced by activin A and Smad2-C induced apoptosis. Activin A converted AR42J into insulin-secreting cells in the presence of hepatocyte growth factor and introduction of Smad2-N inhibited the differentiation. Smad2-C, however, did not induce differentiation in the presence of hepatocyte growth factor. CONCLUSIONS/INTERPRETATION: Activation of the Smad2 pathway is necessary and sufficient to induce apoptosis and morphological changes. Although activation of the Smad2 pathway is required, it is not solely sufficient for the differentiation of AR42J into endocrine cells. PMID- 10382593 TI - Differential accumulation of advanced glycation end products in the course of diabetic retinopathy. AB - AIMS/HYPOTHESIS: Glycated proteins, formed by reaction of glucose and protein, react further yielding numerous, mostly undefined advanced glycation end products (AGE). The recently characterized imidazolone-type AGE (AG-1) is non-oxidatively formed involving 3-deoxyglucosone whereas some AGEs, particularly N(epsilon) (carboxymethyl)lysine (CML), are formed only in the presence of oxygen. METHODS: To study the possible contribution of oxidative and non-oxidative AGE formation in the development of diabetic retinopathy antibodies directed against CML-type and imidazolone-type AGEs were characterized by dot blot analysis and used to localize these well-characterized epitops in the retinas from diabetic rats (early course) and from human Type I (insulin-dependent) diabetes mellitus with laser-treated proliferative diabetic retinopathy (late course). RESULTS: In non diabetic rats CML was moderately positive in neuroglial and vascular structures of non-diabetic rat retinas and increased strongly in diabetic retinas. Anti imidiazolone antibody staining was strongly positive only in diabetic capillaries. Advanced human diabetic retinopathy showed strong CML immunolabelling of the entire retina whereas control samples showed moderate staining of neuroglial structures only with the polyclonal CML-antibody. Anti imidiazolone antibody staining was faint in the inner retina of control sections but were strong throughout the entire diabetic retina. Immunolabelling for the AGE-receptor was congruent with a marker of Muller cells. CONCLUSION/INTERPRETATION: Our data indicate that the oxidatively formed CML is present in non-diabetic retinas as a regular constituent but increases in diabetes both in neuroglial and vascular components. Imidazolone-type AGE are restricted to microvessels and spread during later stages over the entire retina, co-localizing with the expression of AGE-receptor. PMID- 10382594 TI - A new minimally invasive technique to show nerve ischaemia in diabetic neuropathy. AB - AIMS/HYPOTHESIS: Experimental studies have shown that abnormalities of nerve microcirculation are important factors in the pathogenesis of diabetic neuropathy but there have been few clinical studies. We have applied microlightguide spectrophotometry to measure intravascular oxygen saturation (HbO2%) and blood flow in human sural nerve. METHODS: We studied ten patients with mild-moderate sensory motor diabetic neuropathy, nine patients without neuropathy and nine control subjects. We took 300 measurements of oxygen saturation under direct visual control through a 1.9 mm rigid endoscope over three regions of the nerve. Spectrophotometric measurements of nerve fluorescence were taken after an intravenous injection of sodium fluorescein and the rate of increase in nerve fluorescence (rise time) was used as an indicator of nerve blood flow. RESULTS: Nerve oxygen saturation was reduced in patients with neuropathy compared with control subjects (67.1 +/- 2.2% vs 76.7 +/- 2.1%, p = 0.006). Fluorescein rise time was prolonged in patients with neuropathy compared with the control group (48.5 +/- 7.0 s vs 14.0 +/- 3.1 s, p = 0.001) suggesting impaired nerve blood flow. There was a correlation between rise time, nerve oxygen saturation, glycaemic control and sural nerve sensory conduction velocity (p < 0.01). CONCLUSION/INTERPRETATION: The combination of microlight-guide spectrophotometry and micro-endoscopy provides a valuable minimally invasive technique for clinical investigation of nerve microcirculation. We have shown reduced nerve oxygenation and impaired blood flow in diabetic neuropathy and these findings strongly support a central role of microvascular disease in the pathogenesis of diabetic neuropathy. PMID- 10382595 TI - Only limited effects of aminoguanidine treatment on peripheral nerve function, (Na+,K+)-ATPase activity and thrombomodulin expression in streptozotocin-induced diabetic rats. AB - AIMS/HYPOTHESIS: Aminoguanidine, a potent anti-glycation reagent, is known to be beneficial in experimental diabetic neuropathy. In this study, we explored the mechanisms of how aminoguanidine inhibits neuropathic changes in diabetes and compared its effects with those of insulin treatment. METHODS: Wistar rats, aged 8 weeks, were made diabetic by streptozotocin and given aminoguanidine dissolved in drinking water (1 g/l) for 8 weeks. Effects of daily insulin (protamine-zinc) treatment were also examined for comparison. At the end of the 8 weeks, we examined the peripheral nerve function and (Na+,K+)-ATPase activity and their relation to serum thrombomodulin concentrations that are considered as a marker of endothelial injury. RESULTS: Aminoguanidine treatment reduced the diabetes induced decrease in tibial nerve conduction velocity by 47% (p < 0.05 vs untreated diabetic rats) and inhibited the loss of sciatic nerve (Na+,K+)-ATPase activity by 54% (p < 0.05 vs untreated diabetic rats). Insulin-treatment of diabetic rats restored these variables by 83% and 75%, respectively (both, p < 0.01 vs untreated diabetic rats). Thrombomodulin concentrations were increased (p < 0.01) in diabetic rats compared with those in non-diabetic controls and unaffected by aminoguanidine treatment. In contrast, the concentrations remained within the normal range in the insulin-treated group. CONCLUSION/INTERPRETATION: Although aminoguanidine treatment improved nerve conduction velocity and (Na+,K+) ATPase activity, its effects were considerably less than those of insulin and were not apparent in some measures of endothelial cell injury. PMID- 10382596 TI - The role of nitric oxide synthase isoforms and arginase in the pathogenesis of diabetic foot ulcers: possible modulatory effects by transforming growth factor beta 1. AB - AIMS/HYPOTHESIS: L-arginine, an amino acid involved in wound healing, is metabolised by one of two pathways; nitric oxide synthase and arginase. If metabolised by nitric oxide synthase, this can result in tissue destruction, or matrix deposition if metabolised by arginase. The aim therefore was to investigate the role of these enzymes in the pathogenesis of diabetic foot ulcers. METHODS: The activity, proteins by Western blot analysis and cellular distribution (using immunocytochemistry) of these enzymes were measured in diabetic foot ulcers, diabetic skin and normal skin. RESULTS: Total and inducible nitric oxide synthase (p < 0.001) and endothelial nitric oxide synthase were increased in diabetic ulcers compared with diabetic and normal skin and were associated with increased plasma nitrite concentrations in diabetic ulcers (p < 0.05). Inducible nitric oxide synthase was the major isoform, with the macrophage being the predominant cellular source. Similarly arginase activity was increased (p < 0.01) in diabetic ulcers. The protein levels corroborated with the activity data, with the fibroblast being the major cellular source. The spatial and cellular distribution of the two enzyme systems was distinct. Transforming growth factor-beta1 was decreased in diabetic ulcers in comparison with diabetic skin and normal skin. CONCLUSION/INTERPRETATION: Increased nitric oxide synthase activity in diabetic foot ulcers may be responsible for the impaired healing in this disease. Furthermore, the increased activity of arginase could account for the characteristic callus formation around these ulcers. In addition, the lower concentrations of transforming growth factor-betal in diabetic ulcers may explain the raised concentrations of nitric oxide in this condition. PMID- 10382597 TI - The PPARgamma2 amino acid polymorphism Pro 12 Ala is prevalent in offspring of Type II diabetic patients and is associated to increased insulin sensitivity in a subgroup of obese subjects. AB - AIMS/HYPOTHESIS: Recently a mutation in the coding sequence of the adipocyte specific isoform peroxisome proliferator-activated receptor gamma2 (PPARgamma2) was described, leading to the substitution of Proline to Alanine at codon 12. Mutations in PPARgamma2 could have a role in people who are at increased risk for the development of obesity and Type II (non-insulin-dependent) diabetes mellitus. METHODS: Non-diabetic first-degree relatives (n = 108) of subjects with Type II diabetes were characterized by oral glucose tolerance tests and euglycaemic hyperinsulinaemic glucose clamp to determine insulin sensitivity. RESULTS: We found 75 (69%) probands without the PPARgamma ProAla12 substitution, 28 heterozygotes (26%) and 5 (4%) homozygotes. When the whole group was analysed for an association between the mutation and insulin sensitivity, no statistical significance could be shown. Only in the group with severe obesity more than 30 kg/m2, an association (p = 0.016) of the polymorphism with an increase in insulin sensitivity was found. CONCLUSION/INTERPRETATION: These observations suggest that the mutation in the PPARgamma2 molecule may have a role in subgroups prone to the development of obesity and Type II diabetes. PMID- 10382599 TI - Identification of two nonglycosylated polypeptides of Taenia solium recognized by immunoglobulins from patients with neurocysticercosis. AB - An immunoprecipitation technique using biotin-labeled proteins of Taenia solium was developed to identify antigens recognized by immunoglobulins from patients with neurocysticercosis. Six major polypeptides of 100, 70, 50, 42, 35, and 24 kDa were recognized by cerebrospinal fluid from most serologically positive patients. All polypeptides except the 70- and 35-kDa antigens were retained on a lentil-lectin chromatography column and were recognized by lentil lectin in an overlay assay. The 70- and 35-kDa antigens were not labeled with biotin hydrazide, indicating that saccharide residues are not present in these two polypeptides. Furthermore, the 70- and 35-kDa antigens were recognized by antibodies of more than 86% of patients serologically positive for neurocysticercosis as opposed to none of the patients afflicted with other neuropathologies of the central nervous system. This finding indicates that immunodiagnosis of neurocysticercosis can be carried out with antigens different from those used in the standard enzyme-linked immunoelectrotransfer assay. PMID- 10382598 TI - Augmented growth response to IGF-1 via increased IRS-1 in Chinese hamster ovary cells expressing kinase-negative insulin receptors. AB - AIMS/HYPOTHESIS: Although both increased cell growth and impaired insulin signalling have been associated with diabetes, this association has not been investigated. Insulin-like growth factor-1 (IGF-1), a structural and functional analog of insulin, may play a part in the aberrant insulin receptor-mediated signalling observed in diabetes. METHODS: To investigate the consequence of this impaired signalling on cell proliferation and transformation, we transfected Chinese hamster ovary cells with cDNA encoding a kinase-defective insulin receptor. RESULTS: In these mutant cells, the mitogenic and metabolic effects of insulin were reduced compared with control cells (p < 0.05) and this was due to a dominant negative effect. In contrast, these mutant cells showed a higher mitogenic response to IGF-1 than control cells, although IGF-1 receptor expression was similar in both cell lines. There was no statistically significant difference in mitogenic response, however, to platelet-derived growth factor, basic fibroblast growth factor and heparin-binding epidermal growth factor-like growth factor. Variables of the IGF-1 signalling pathway, including tyrosine phosphorylation of insulin receptor substrate-1 and activation of mitogen activated protein kinase and phosphatidyl inositol 3 kinase, were also augmented in mutant cells. Insulin receptor substrate-1 message and protein abundance were higher in mutant than in control cells. Moreover, mutant cells had a higher mitogenic potential in low-serum-containing medium, suggesting that these cells have a transformed phenotype. CONCLUSION/INTERPRETATION: These findings suggest that an impaired insulin signalling may upregulate insulin receptor substrate-1 and that this, in turn, leads to increased IGF-1 signalling, a phenomenon that is possibly associated with increased cell growth in diabetes. PMID- 10382600 TI - Cryptosporidium oocysts in Bent mussels (Ischadium recurvum) in the Chesapeake Bay. AB - Filter-feeding molluscan shellfish can concentrate environmentally derived waterborne pathogens of humans, which can be utilized in the sanitary assessment of water quality. In the present study, oocysts of Cryptosporidium were detected in Bent mussels (Ischadium recurvum) at two Chesapeake Bay sites from which C. parvum-contaminated oysters had previously been collected. Spiking of Cryptosporidium-free blue mussel (Mytilus edulis) tissue with C. parvum oocysts showed a 51.1% recovery rate of oocysts, giving an oocyst detection limit of 19 oocysts/0.7 ml of mussel tissue homogenate. The results indicate that Bent mussels, which are common throughout the Chesapeake Bay region, may prove to be useful as biological indicators of water contamination with Cryptosporidium oocysts. PMID- 10382601 TI - Strongyloides ratti: additive effect of testosterone implantation and carbon injection on the susceptibility of female mice. AB - A sex-related difference in host susceptibility to Strongyloides ratti was previously known. Male mice were more susceptible to S. ratti infection and the difference was seen against migrating larvae under the regulation of testosterone. Against migrating larvae, macrophages were assumed to play important roles in host natural immunity. On the basis of these findings, to examine the effect of testosterone on macrophages we treated female mice with testosterone and/or carbon to block the function of macrophages. Mice were then infected with third-stage larvae of S. ratti. By counting of the migrating larvae in the cranial cavity at 36 h after infection the effect of each treatment was assayed. Testosterone treatment alone (Te) or carbon injection alone (Ca) effectively increased the worm recovery. Given together, Te and Ca (Te + Ca) significantly increased the worm recovery to levels almost equal to the sum of those achieved with Te and Ca. The serum testosterone concentration was elevated in mice that had undergone Te and Te + Ca at the time of worm recovery. Surprisingly, the serum testosterone concentration reached after Te + Ca was elevated more than that attained by Te. The same experiment with a half-dose of Te and Ca (Te half + Ca) resulted in the same testosterone concentration achieved with Te and resulted in a worm recovery almost equal to the sum of that achieved with Te and Ca. These results clearly showed that Te and Ca had an additive effect on the recovery of migrating S. ratti larvae. Testosterone had an effect after macrophages had been blocked. The relationship between testosterone and macrophage function during S. ratti infection is discussed. PMID- 10382602 TI - Review: Theileria schizonts induce fundamental alterations in their host cells. AB - The sporozoites of Theileria annulata and T. parva invade bovine leukocytes, where they differentiate into schizonts. The latter can immortalize and induce fundamental changes in their host cells. T. annulata infects mainly major histocompatibility complex class II cells, whereas T. parva preferentially transforms T-lymphocytes, which proliferate continuously without the need for exogenously added growth factors. Most of the cell lines appear to be independent of a growth factor but may express several cytokines that influence the outcome of the disease. The mechanisms underlying this transformation are not well understood. The infected cells show increased activity of casein kinase II and Jun NH2-terminal kinase (JNK), whereas extracellular signal-related kinase 1 and 2 and P38 are not activated. In addition, several transcriptional factors such as NFkB and AP-1 are activated. It has been postulated that parasite proteins either expressed on the surface of the schizonts or secreted into the host cell cytoplasm may interfere with the signal-transduction pathway of the host cells. A possible candidate may the casein kinase II homologue that was identified in schizonts of both T. annulata and T. parva. PMID- 10382603 TI - Review: the cellular basis of the immunity to and immunopathogenesis of tropical theileriosis. AB - The intracellular protozoan parasite Theileria annulata causes a severe and often fatal disease of pure and crossbred cattle in tropical and subtropical countries. Animals that recover from the infection are immune against challenge with homologous parasite strains. In the present review we refer to the role of immunocompetent cells and their products in containing the infection or in facilitating the progress of the disease. Parasite-infected host cells produce cytokines, which, depending on their concentration and timing of production, may enhance the establishment of the infection. Thus, cell lines producing high levels of proinflammatory cytokines cause severe postvaccinal reactions when inoculated into cattle. This may be supported by an aberrant non-specific activation of naive T-cells, leading to the production of high levels of gamma interferon (IFN-y). Under these circumstances development of the specific immune response may be inhibited. At this stage, innate immune reactions are operating to contain the infection. Natural killer cells and macrophages may represent the most important part of this immunity. Antibodies and specific T-lymphocytes, CD4+ T-cells and cytotoxic T-lymphocytes (CTLs), play the most important role in a challenge infection. In this context, CD4+ T-cells produce cytokines required for the clonal expansion of CTLs that kill their target cells in a major histocompatibility complex (MHC) class I-restricted manner. In addition, CD4+ T cells produce macrophage-activating cytokines such as IFN-gamma. Such activated macrophages produce mediators such as NO, which destroy the intracellular schizonts. Attempts have been directed toward the identification of parasite antigens involved in the induction of immunity. To date, only a limited number of sporozoite and merozoite antigens have been identified and examined for their immunogenicity, and the protection achieved is partial. An effective vaccine must include schizont proteins, notably, those proteins that are secreted into the host cell cytoplasm because these may have access to the MHC class I and II compartments to be presented to CTLs and CD4+ T-cells, respectively. Several schizont proteins have been identified and these are now under investigation. PMID- 10382604 TI - Pitfalls in the application of enzyme-linked immunoassays for the detection of circulating trypanosomal antigens in serum samples. AB - The experimental infection of two goats with Trypanosoma vivax trypanosomes provided samples for analysis using parasitology techniques and antigen-detection enzyme-linked immunosorbent assays (ELISAs) for T. vivax, T. congolense and T. brucei. Clinical, parasitological and serological findings were monitored during the course of infection to identify problems in the application of these ELISAs. The data clearly showed that the ELISAs examined were entirely unsuitable for the reliable detection of trypanosomal antigen. Consequently, research strategies pertinent to the development of a new generation of both antigen and antibody ELISAs are outlined considering the problems encountered. These were (1) the reactivity of the reagents; (2) the specificity of the reagents; (3) the nature of the test sample, e.g. the compartmentalisation of trypanosomes between plasma, serum and red blood cells; (4) possible interference with the ELISA through immune complexing; and (5) the biology of the host/trypanosome relationship to gain an understanding of fluctuations in trypanosomes in the systemic circulation. PMID- 10382605 TI - Influence of the genetic background and parasite load of mice on the immune response developed against nymphs of Ixodes ricinus. AB - The immune response of BALB/c (H-2d), DBA (H-2d), C57BL/6 (H-2b), C3H (H-2k), CBA (H-2k), SJL (H-2s), and FVB (H-2q) mice infested once with 15 nymphs of Ixodes ricinus is polarized toward Th2 as suggested by cytokines produced by lymph node cells stimulated with concanavalin A. The parasite load does not influence the polarization of the immune response as observed in BALB/c mice, which developed a Th2 response when infested with 5 or 45 nymphs. As assessed by attachment and weights of engorged nymphs, no resistance was acquired by BALB/c, C57BL/6, or C3H mice undergoing three successive infestations. However, these mice produced a gradual increase in IgE. PMID- 10382606 TI - The morphology of Ichthyophonus sp. in their mugilid hosts (Pisces: Teleostei) and following cultivation in vitro. A light and electron microscopy study. AB - The morphology of Ichthyophonus sp., a parasite of Mugil capito and Liza saliens, was investigated by light and transmission electron microscopy. The most frequent stage found in the fish hosts was the multinucleate spore, though germinating stages, hyphae, and endospores were also found. Different development patterns were observed in the media assayed for in vitro culture. Optimal growth and development were obtained in Eagle's minimum essential medium (MEM) supplemented with 10% fetal bovine serum at pH 7. Ultrastructural features of multinucleate spores, both in the fish host and in culture, were a fibrillar thick wall and an electron-lucent matrix, with large glycogen granules, some electron-dense bodies, large vacuoles, lipid inclusions, and endoplasmic reticulum mainly appearing among the nuclei. Mitochondria with scarce tubulovesicular cristae were observed in the different stages, mainly near the wall and the germinating sites. Condensed heterochromatin was rarely seen. A nucleus-associated organelle (NAO) was frequently observed, and dictyosome cisternae and vesicles appeared in its vicinity. PMID- 10382607 TI - Lectin-binding properties of different Leishmania species. AB - Carbohydrate cell-surface residues on stationary promastigotes of 19 isolates of Leishmania were studied with a panel of 27 highly purified lectins, which were specific for N-acetyl-D-glucosamine, D-mannose, L-fucose, D-galactose, N-acetyl-D galactosamine, and sialic acid. The specificity of the cell-surface carbohydrates was analyzed by agglutination and radioiodinated lectin-binding assays. L. (L.) amazonensis and L. (L.) donovani were agglutinated by 12 and 10 of the 27 lectins used, respectively. Artocarpus integrifolia lectin (Jacalin) was incapable of agglutinating the tested species of the donovani complex, and this result was confirmed by radioiodinated Jacalin-binding assays. Jacalin had an average of 3.8 x 10(6) receptors/L. (L) amazonensis promastigote and bound with an association constant of 5 x 10(6) M(-1). PMID- 10382608 TI - Vismione H and structurally related anthranoid compounds of natural and synthetic origin as promising drugs against the human malaria parasite Plasmodium falciparum: structure-activity relationships. AB - Natural and synthetic anthranoid compounds were subjected to an evaluation against asexual erythrocytic stages of the human malaria parasite Plasmodium falciparum in vitro. Stimulated by the good activities of Vismia guineensis extracts, a more detailed investigation of the active principles revealed the pre nylated preanthraquinone vismione H (1) to be a potent antimalarial [50% growth inhibitory concentration (IC50) 0.088 microg/ml]. On the basis of this finding a series of chemically related phlegmacins (2-5), flavomannins (6-8), and rufoolivacins (9-11) isolated from several species of Cortinarius, a genus of higher fungi, and 5 synthetic vismione-like anthranoids (12-16) were evaluated as well. Although these compounds displayed weaker antiplasmodial effects than did vismione H (1) itself, considerable levels of activity were obtained with phlegmacin B1 (2), flavomannin-6,6'-di-O-methyl ether A1 (6), trans-4-hydroxy flavomannin-6,6'-di-O-methyl ether A (8), and rufoolivacin B (10). Initial preconditions for activity within the vismiones and related anthranoids were established. PMID- 10382609 TI - Oviposition of Lymnaea truncatula infected by Fasciola hepatica under experimental conditions. AB - Experimental infections of Lymnaea truncatula by Fasciola hepatica (one, two, or three miracidia per snail) were carried out under laboratory conditions to analyze the oviposition of infected snails and determine the characteristics of their egg masses. In the infected snails from the three groups, egg-laying steadily decreased until week 4 postexposure and stopped afterward until the end of the experiment, except for the cercaria-shedding snails from the one miracidium group, for which low numbers of egg masses were noted between weeks 9 and 12. In uninfected snails the number of egg masses decreased until week 4 and remained low during subsequent weeks. At weeks 11 and 12 postexposure the natality rate was 98.9% in controls, 56% in the cercaria-shedding snails from the one-miracidium group, and 59.2-68.5% in uninfected snails. In the cercaria shedding snails from the one-miracidium group the restoration of reproduction activity after week 8 may be explained by a lower parasite burden in these snails than in those from the two- and three-miracidium groups. PMID- 10382610 TI - Immunosuppression in hamsters with progressive visceral leishmaniasis: an evaluation of the role of nitric oxide toward impairment of the lymphoproliferative response. AB - The progressive visceral infection caused in golden hamsters by Leishmania donovani amastigotes led to gradual impairment of the proliferative response of their splenic (SPMC) or peripheral blood (PBMC) mononuclear cells to in vitro stimulation with leishmanial antigen, with mitogen (concanavalin A), and even with a combination of phorbol myristate acetate (PMA) and ionomycin (Io). Removal of macrophage-like adherent cells from SPMC or PBMC of infected animals, however, almost completely restored their proliferative response to PMA + Io, thus ruling out the possibility of any intrinsic defect in the signal-transduction pathways of lymphocyte activation and proliferation. Subsequent studies demonstrated that the generation of soluble mediators such as nitric oxide by these adherent cells is responsible, albeit partially, for the down-regulation of the lymphoproliferative response in hamsters with visceral leishmaniasis. PMID- 10382611 TI - A new nematode, Ansiruptodera scapteromi sp.nov. (Nematoda: Aspidoderidae), recovered from the Argentinean water rat Scapteromys tumidis (Waterhouse, 1837) in Uruguay. AB - This report describes a new species of aspidoderid nematode, Ansiruptodera scapteromi sp. nov., the second species to be reported in the genus Ansiruptodera Skrjabin and Shikhobalova, 1947. The A. scapteromi sp. nov. is the first species of the genus to be recorded from a rodent host. The new species is clearly different from the only other species, A. ansiruipta (Proenca, 1937) Skrjabin and Shikhobalova, 1947, in that it possesses short lateral alae that terminate before the midbody; a smaller cephalic extremity; a shorter esophagus, pharynx, tail, and tail appendage; a smaller sucker; and longer spicules. The two species also differ in the numbers and arrangements of caudal papillae. A. scapteromi appears to be a parasite of capture and the water rats seem to have been infected from Edentata in Uruguay. PMID- 10382612 TI - Isolation and identification by partial sequencing of the 18S ribosomal gene of free-living amoebae from necrotic tissue of Basilliscus plumifrons (Sauria: Iguanidae). AB - A 3-year-old Basiliscus plumifrons developed a necrotic lesion on the tail resulting from nodules of unknown etiology. Investigation of necrotic tissue revealed several gram-negative bacteria as well as three different species of free-living amoebae. The amoebae were identified by morphological characters as belonging to the genera Acanthamoeba, Echinamoeba, and Naegleria, respectively. Partial sequencing of the 18S ribosomal gene was performed for reliable systematic determination. Two of the isolates showed thermotolerance. No isolate was growable in conventional liquid media, but the Acanthamoeba strain readily grew on a human cell line (HEp2). It remains unclear whether the amoebae fed on the coexisting bacteria or on host tissue. PMID- 10382613 TI - DNA polymerase activity in encysting Entamoeba invadens. AB - Using an axenic encystation system of Entamoeba invadens as a model for E. histolytica encystation, we examined the level of DNA polymerase activity in E. invadens during encystation induced in vitro. We first characterized the DNA polymerase activity of trophozoites of E. invadens, comparing it with that of E. histolytica, and found that the activity of E. invadens was lower than that of E. histolytica at pH 2, 4, and 6 and was higher at pH 8 and 10. The activity of E. invadens was completely inhibited by high concentrations of K(-). Among inhibitors of mammalian DNA polymerases, aphidicolin and N-ethylmaleimide inhibited the activity, but 2',3'-dideoxythymidine-5'-triphosphate did not. Thus, the sensitivity of the E. invadens activity to salt and inhibitors of mammalian DNA polymerases was basically the same as that recorded for E. histolytica in our previous results. The level of DNA polymerase activity in cysts decreased as encystation proceeded as compared with that of trophozoites. The results indicate that encystation is accompanied by a reduced level of DNA polymerase activity, which correlates with the previous finding that nuclear division occurs during cyst maturation in the absence of DNA synthesis. PMID- 10382614 TI - Cloning and characterization of Rab and Ran/TC4 cDNA clones in Trichinella spp. AB - The cloning and characterization of seven Rab and three Ran/TC4 partial cDNA sequences in both cystic (Trichinella spiralis and T. britovi) and noncystic species (T. pseudospiralis) are reported. These molecules were cloned by rapid amplification of cDNA ends via polymerase chain reaction (RACE-PCR), using cDNA from the aforementioned Trichinella spp. coupled to the AP1 adaptor. As primers, AP1 and 5B (derived from the WDTAGQE sequence of region 2 specific for Rab and Ran proteins) sequences were included in the PCR. The cloned cDNAs were sequenced and characterized by both Southern-blot and Northern-blot analysis. Trichinella spp. Rab- and Ran-like molecules showed divergences in both the nucleotide and the deduced amino acid sequences as compared with the corresponding homologues previously described in other organisms. In addition, differences were observed among the Trichinella species, mainly between the cystic and the noncystic species, in both DNA restriction-enzyme polymorphism and expression of the six GTPases isolated. PMID- 10382615 TI - Cytokine gene expression following neural grafts. AB - A reverse-transcription polymerase chain reaction study of molecular events following solid neural tissue xenogeneic and syngeneic grafts into the mouse caudate nucleus was made between 1 day and 30 days post-grafting. Constitutive expression of interleukin (IL)-1 and transforming growth factor (TGF)-beta1 mRNAs was detected. The sham operation produced minimal cytokine upregulation, indicating that surgical trauma caused minimal damage. A similar picture was seen with syngeneic grafts, which showed good graft survival. Xenografts, however, were rejected within 30 days and cytokine mRNAs for IL-2, IL-2R, IL-4, IL-10 and interferon (IFN)-gamma were detected from 7 days post-graft, correlating with histological identification of a leukocytic infiltrate. This method provides a quick, comparative screen for detecting cytokine mRNA profiles in neural grafts and may assist the diagnosis of early rejection phenomena. As such, it is a potential analytical tool for strategies aimed at depleting/blocking the activity of different cell types and/or cytokines in future neural transplant therapy. PMID- 10382616 TI - The uncontrolled manifold concept: identifying control variables for a functional task. AB - The degrees of freedom problem is often posed by asking which of the many possible degrees of freedom does the nervous system control? By implication, other degrees of freedom are not controlled. We give an operational meaning to "controlled" and "uncontrolled" and describe a method of analysis through which hypotheses about controlled and uncontrolled degrees of freedom can be tested. In this conception, control refers to stabilization, so that lack of control implies reduced stability. The method was used to analyze an experiment on the sit-to stand transition. By testing different hypotheses about the controlled variables, we systematically approximated the structure of control in joint space. We found that, for the task of sit-to-stand, the position of the center of mass in the sagittal plane was controlled. The horizontal head position and the position of the hand were controlled less stably, while vertical head position appears to be no more controlled than joint motions. PMID- 10382618 TI - Rule-dependent neuronal activity in the prefrontal cortex. AB - We studied single-neuron activity in the prefrontal cortex (PF) while a monkey performed a task according to two different rules, termed conditional and spatial. The monkey viewed a video screen, and its task required a hand movement in response to the dimming of a light spot. There were four light spots on the screen: right, left, up, and down from the center. Only one of the four spots dimmed, and the degree of dimming was slight. Accordingly, the monkey needed to foveate the "correct" light spot to detect the dimming. A visual cue indicated which of the four light spots would be deemed correct and, thus, would dim on each trial. The sequence of events was as follows: a fixation spot appeared at the center of the screen; then, a cue appeared twice at one of the four potential target locations; then, the four target spots appeared; and, finally, one of them dimmed. Except for the color of an initial fixation point, the cues, their locations, and other events were identical for the conditional and spatial rules. The rules differed in one essential way. For the conditional rule, nonspatial attributes of the visual cue indicated which of the four light spots would dim, and the cue's location was irrelevant. For the spatial rule, the cue's location determined the correct target on that trial. The light spot at the location of the cue always dimmed, regardless of which cue appeared there. Our sample included 221 PF neurons showing significant task-related activity modulation, distributed among dorsal, dorsolateral, and ventral PF regions. Between one-third and one-half of the sample in each of those regions showed statistically significant activity differences that could be attributed to the rule. Selectivity for cues and/or their locations was common. However, there was no significant regional segregation of such selectivity. These data support the hypothesis that PF plays a role in the guidance of behavior according to previously learned rules. PMID- 10382617 TI - Ground reaction forces in locomoting hemi-parkinsonian rats: a definitive test for impairments and compensations. AB - Hemi-parkinsonian rats have preserved postural reflexes but are impaired in initiation of voluntary movements. Surprisingly, these rats can walk and run, suggesting that they can access some compensatory strategy to overcome the rigidity in their impaired limbs. The purpose of the present experiment was to investigate the locomotor compensations made by hemi-parkinsonian rats by measuring the forces exerted by the limbs on the ground throughout the stride during trotting. Rats with unilateral dopamine depletion produced by injection of 6-hydroxydopamine into the nigrostriatal bundle were trained to run back and forth in an alley for food reinforcement. Ground reaction forces were measured in three orthogonal directions using a force plate embedded in the runway. Rats were also videotaped so that limb movements were synchronized with force recordings. Although locomotion was obviously impaired, the affected limbs could support weight and provide some braking forces. In addition, the impaired hindlimb provided significant propulsive force, and a relatively large laterally directed force. Analysis of vertical movement of the centre of mass suggested that the impaired hindlimb was being used partly as a spring. The most significant abnormalities were seen during the diagonal couplet of the impaired forelimb and the unimpaired hindlimb, partly reflecting the important compensatory role of the unimpaired hindlimb. These results demonstrate that this method is useful in the analysis of hemi-parkinsonian gait and provide insights as to how rats can use an impaired limb to produce weight support and propulsion. PMID- 10382619 TI - Forebrain connections of medial agranular cortex in the prairie vole, Microtus ochrogaster. AB - Fluorescent axonal tracers were used to investigate the connections of medial agranular cortex (frontal area 2, Fr2) in male prairie voles. The rostral and caudal portions of Fr2 (rFr2 and cFr2) have distinct but partially overlapping patterns of connections. Thalamic labeling after cFr2 injections was present in anteromedial nucleus (AM), ventrolateral nucleus (VL), lateral segment, mediodorsal nucleus (MDl), centrolateral nucleus (CL), ventromedial nucleus (VM), posterior nucleus (Po) and lateral posterior nucleus (LP). A band of labeled cells involving CL, central medial nucleus (CM) and rhomboid nucleus (Rh) formed a halo around the periphery of submedial (gelatinosus) nucleus (Sm). Within cFr2 there is a rostrocaudal gradient whereby projections from VL and MDl become progressively sparser caudally, whereas those from LP and Po become denser. Rostral Fr2 receives afferents from a similar group of thalamic nuclei, but has denser innervation from VL and MDl, lacks afferents from LP, and receives less input from nuclei around the periphery of Sm. Caudal Fr2 has extensive cortical connections including orbital cortex, rostral Fr2, Fr1, caudal parietal area 1 (Par1), parietal area 2 (Par2), and posterior parietal, retrosplenial and visual areas. Rostral Fr2 has similar connections with areas Fr1, Par1 and Par2; orbital connections focused in ventrolateral orbital cortex (VLO); connections with caudal Fr2; greatly reduced connections with posterior parietal cortex and the visual areas; and no connections with retrosplenial cortex. The axons linking rFr2 and cFr2 with each other and with other cortical areas travel predominately in the deep gray matter of layers VI and VII rather than in the white matter. Projections to the dorsal striatum from rFr2 are widespread in the head of the caudate, become progressively restricted to a dorsocentral focus more caudally, and disappear by the level of the anterior commissure. The projections from cFr2 are largely restricted to a focal dorsocentral region of the striatum and to the dorsolateral margin of the caudatoputamen. In comparison to area Fr2, the laterally adjacent area Fr1 has thalamic and cortical connections which are markedly restricted. Area Fr1 receives thalamic input from nuclei VL, anteroventral nucleus (AV), CL and Po, but none from mediodorsal nucleus (MD) or LP, and its input from VM is reduced. Cortical afferents to Fr1 originate from areas Fr2, caudal Par1 and Par2. Medial agranular cortex of prairie voles has a pattern of connections largely similar to that seen in rats, suggesting that area Fr2 in prairie voles is part of a cortical network that may mediate complex behaviors involving spatial orientation. PMID- 10382620 TI - Effects of electrical stimulation of the tooth pulp and phrenic nerve fibers on C1 spinal neurons in the rat. AB - Effects of electrical stimulation of the ipsilateral tooth pulp (TP) on C1 spinal neurons were determined in 33 anesthetized rats. One hundred and seven neurons responded to TP stimulation. In 10 rats, the activity of 18 C1 spinal neurons and the amplitude of a digastric electromyogram (dEMG, n = 10) increased proportionally during the TP stimulation at an intensity of 1-3 times the threshold for jaw-opening reflex (JOR). Excitatory receptive somatic fields were examined in 61 neurons. Somatic field locations of many neurons (67.2%) involved the ipsilateral face, neck, and jaw. The activity of 45 neurons was increased by both noxious pinch and brushing hair. Of the 107 C1 spinal neurons responding to TP stimulation, 55 were tested to determine the effects of electrical stimulation of the ipsilateral phrenic nerve (PN) above the heart. Twenty-eight of 55 neurons tested were excited; no change in activity was seen for the remaining 27 neurons. The activity of six neurons increased as the intensity of PN stimulation was increased. Excitatory receptive somatic fields were determined in 28 neurons, and somatic field locations of 17 neurons (60.7%) included the ipsilateral face, neck, and jaw. Both noxious pinch and brushing hair excited all 28 neurons. These results suggest that there may be the convergence of face, neck, jaw, TP, and PN afferents on the same C1 spinal neurons in the rat. PMID- 10382621 TI - Adaptation of the walking pattern to uphill walking in normal and spinal-cord injured subjects. AB - Lower-limb movements and muscle-activity patterns were assessed from seven normal and seven ambulatory subjects with incomplete spinal-cord injury (SCI) during level and uphill treadmill walking (5, 10 and 15 degrees). Increasing the treadmill grade from 0 degrees to 15 degrees induced an increasingly flexed posture of the hip, knee and ankle during initial contact in all normal subjects, resulting in a larger excursion throughout stance. This adaptation process actually began in mid-swing with a graded increase in hip flexion and ankle dorsiflexion as well as a gradual decrease in knee extension. In SCI subjects, a similar trend was found at the hip joint for both swing and stance phases, whereas the knee angle showed very limited changes and the ankle angle showed large variations with grade throughout the walking cycle. A distinct coordination pattern between the hip and knee was observed in normal subjects, but not in SCI subjects during level walking. The same coordination pattern was preserved in all normal subjects and in five of seven SCI subjects during uphill walking. The duration of electromyographic (EMG) activity of thigh muscles was progressively increased during uphill walking, whereas no significant changes occurred in leg muscles. In SCI subjects, EMG durations of both thigh and leg muscles, which were already active throughout stance during level walking, were not significantly affected by uphill walking. The peak amplitude of EMG activity of the vastus lateralis, medial hamstrings, soleus, medial gastrocnemius and tibialis anterior was progressively increased during uphill walking in normal subjects. In SCI subjects, the peak amplitude of EMG activity of the medial hamstrings was adapted in a similar fashion, whereas the vastus lateralis, soleus and medial gastrocnemius showed very limited adaptation during uphill walking. We conclude that SCI subjects can adapt to uphill treadmill walking within certain limits, but they use different strategies to adapt to the changing locomotor demands. PMID- 10382622 TI - Immunoelectron microscopic study of gamma-aminobutyric acid inputs to identified thalamocortical projection neurons in the anterior thalamus of the rat. AB - We have carried out a semi-quantitative ultrastructural study to determine the characteristics and distribution of gamma-aminobutyric acid (GABA)-containing constituents of the anterodorsal (AD) and anteroventral (AV) thalamic nuclei in adult rats. We used a polyclonal antibody to GABA and a postembedding immunogold detection method in animals in which the cortical projection neurons of these nuclei had been labelled by retrograde transport of cholera toxin/horseradish peroxidase (HRP) injected into the retrosplenial granular cortex. Two types of GABA-immunopositive structures were identified, with gold particle densities 4-40 times higher than the highest densities over blood-vessel lumens and areas of empty resin: (1) an apparently homogeneous population of axon terminals with Gray type-2 (symmetric) synaptic contacts corresponding to F-axon terminals; and (2) small-medium sized myelinated axons scattered individually or in small groups within the neuropil which may be their parent axons. These axons and terminals may originate from the ipsilateral thalamic reticular nucleus; others may arise from the basal forebrain or brainstem. The GABA-immunopositive terminals comprised approximately 16% of all axon terminal profiles in AD and 12% in AV, a significant difference. However, because the immunoreactive axon terminals in AD were significantly larger than those in AV (1.09+/-0.47 microm2 vs 0.90+/-0.43 microm2) and would therefore be encountered more frequently, it is not possible to conclude that the GABAergic innervation of AD is heavier than that of AV. The GABA-positive terminals established synaptic contacts with cell bodies and dendrites of all sizes (some of which were HRP-labelled) with the following frequency distribution (AD/AV, no significant difference): somata 5%/7%; large dendrites (> or = 1.5 microm) 14%/9%; medium dendrites (1.00-1.49 microm) 35%/45% and small dendrites (< 1 microm) 46%/40%. Despite evidence from previous studies, we found no evidence in this study for the presence of GABAergic interneurons or for GABA-containing projection neurons in AD or AV. PMID- 10382623 TI - Experimental model of trigeminal pain in the rat by constriction of one infraorbital nerve: changes in neuronal activities in the somatosensory cortices corresponding to the infraorbital nerve. AB - Neuronal activities of the somatosensory (Sm1) vibrissa cortex were explored bilaterally under moderate gaseous anaesthesia in rats with a chronic constriction injury (CCI) of an infraorbital nerve (IoN). The CCI-IoN rats exhibited abnormal pain-related reactions to mechanical stimuli directed to the territory of the injured nerve, just prior to the recording session. The responsiveness, laminar localisation, and somatotopic organisation of 388 neurones were recorded in the pad vibrissa cortex contralateral (Cc, n=249) and ipsilateral (Ci, n=139) to the injured nerve, analysed, and compared with 223 neurones recorded in a parallel study of the Sm1 cortex in normal rats (Cn). The mean background activity of all the recorded neurones was relatively low, whether they were located in the Cn, Cc or Ci. The responsive neurones occurred in similar proportions in the Cc and Ci (approximately 55%) and significantly more frequently than in the Cn (35%). They consisted mainly of vibrissa neurones (100% in Cn, 85% in Cc, and 93% in Ci). The ratio [vibrissa neurones/(vibrissa+unresponsive) neurones] was enhanced in all the cortical layers of the Cc and Ci, except in the Cc layer IV. The receptive field (RF) size of the vibrissa neurones also differed greatly. It was limited to one vibrissa for 88% of the Cn neurones, but expanded to two or more vibrissae for 72% of the Cc and 52% of the Ci neurones. These multivibrissa units, mostly located in layers V and VIa for the few Cn neurones, were scattered in the different layers in CCI-IoN rats, with the largest RFs occurring in the deepest layers. In parallel, the cortical somatotopy of the vibrissae, roughly comparable with that initially described in pioneer studies of normal rats, was dramatically disturbed not only in the Cc, but also in the Ci of CCI-IoN rats. Contrasting with the results previously obtained in the ventro-postero-median thalamic nucleus of CCI IoN rats, no neurones were driven by pinches or pinpricks applied to the cutaneous part of the vibrissa pad. It is questioned whether the disorganisation within the cortical map of the whisker pad, and the expanded RFs of vibrissa neurones could account for the abnormal pain-related reactions elicited from the massively deafferented trigeminal area. PMID- 10382624 TI - Clustering of Pacinian corpuscle afferent fibres in the human median nerve. AB - To further study the functional organisation of human peripheral nerves, the intrafascicular arrangement of afferent fibres supplying Pacinian corpuscles (PCs) was explored by percutaneous microneurography using thin-calibre, concentric needle electrodes. In normal adults, 20 PC afferents were identified in 13 recording sites. Low-amplitude (less than 30 microm) vibratory stimuli to the skin were applied with tuning forks oscillating at 128 Hz or 256 Hz and response patterns of individual PC units were studied. In many recording sites, two, sometimes even three, PC afferents with adjacent or overlapping receptive fields in the hand were clustered in the nerve. The observed incidence in the records containing a certain number of PC units was compared with the expected probability calculated according to the hypothesis that all nerve fibres are randomly organised in peripheral nerves. The results suggested that PC afferents are partially segregated in the nerve. In addition, PC afferents were neighbouring on slowly adapting type II (SAII) units and skin sympathetic activity in individual fascicles. SAII units often innervated the same skin area as PC units, but did not respond to vibration. The data provided additional information regarding the functional organisation of the human peripheral nerve and the mechanisms underlying the sense of vibration in man with special regard to population behaviour of neighbouring PC mechanoreceptors. PMID- 10382625 TI - Saccular and utricular inputs to sternocleidomastoid motoneurons of decerebrate cats. AB - Connections from the otolithic organs to sternocleidomastoid (SCM) motoneurons were studied in 20 decerebrate cats. The electrical stimulation was selective for the saccular or the utricular nerves. Postsynaptic potentials were recorded from antidromically identified SCM motoneurons; these muscles participate mainly in neck rotation and flexion. Partial transections of the brainstem at the level of the obex were performed to identify the possible pathway from the otolithic organs to the SCM motoneurons. Saccular or utricular nerve stimulation mainly evoked inhibitory postsynaptic potentials (IPSPs) in the ipsilateral SCM motoneurons. Some of the sacculus-induced IPSPs were preceded by small-amplitude excitatory PSPs (EPSPs). The latencies of the PSPs ranged from 1.8 to 3.1 ms after saccular nerve stimulation and from 1.7 to 2.8 ms after utricular nerve stimulation, indicating that most of the ipsilateral connections were disynaptic. In the contralateral SCM motoneurons, saccular nerve stimulation had no or faint effects, whereas utricular nerve stimulation evoked EPSPs in about two-thirds of neurons, and no visible PSPs in about one-third of neurons. The latencies of the EPSPs ranged from 1.5 to 2.0 ms, indicating the disynaptic connection. Thus, the results suggest a difference between the two otolithic innervating patterns of SCM motoneurons. After transection of the medial vestibulospinal tract (MVST), saccular nerve stimulation did not evoke IPSPs at all in ipsilateral SCM motoneurons, but some (11/40) neurons showed small-amplitude EPSPs. Most (24/33) of the utricular-activated IPSPs disappeared after transection, whereas the other 9 neurons still indicated IPSPs. In the contralateral SCM motoneurons, no utricular-activated EPSPs were recorded after transection. These MVST transection results suggest that most of the otolith-SCM pathways are located in the MVST at the obex level. However, the results also suggest the possibility that other otolith-SCM pathways exist at the obex level. PMID- 10382626 TI - Kinesthetic perceptions of earth- and body-fixed axes. AB - The major purpose of this research was to determine whether kinesthetic/proprioceptive perceptions of the earth-fixed vertical axis are more accurate than perceptions of intrinsic axes. In one experiment, accuracy of alignment of the forearm to earth-fixed vertical and head- and trunk-longitudinal axes by seven blindfolded subjects was compared in four tasks: (1) Earth-Arm--arm (humerus) orientation was manipulated by the experimenter; subjects aligned the forearm parallel to the vertical axis, which was also aligned with the head and trunk longitudinal axis; (2) Head--head, trunk, and upper-limb orientations were manipulated by the experimenter, subjects aligned the forearm parallel to the longitudinal axis of the head using only elbow flexion/extension and shoulder internal/external rotation; (3) Trunk--same as (2), except that subjects aligned the forearm parallel to the trunk-longitudinal axis; (4) Earth--same as (2), except that subjects aligned the forearm parallel to the earth-fixed vertical. Head, trunk, and gravitational axes were never parallel in tasks 2, 3, and 4 so that subjects could not simultaneously match their forearm to all three axes. The results showed that the errors for alignment of the forearm with the earth-fixed vertical were lower than for the trunk- and head-longitudinal axes. Furthermore, errors in the Earth condition were less dependent on alterations of the head and trunk orientation than in the Head and Trunk conditions. These data strongly suggest that the earth-fixed vertical is used as one axis for the kinesthetic sensory coordinate system that specifies upper-limb orientation at the perceptual level. We also examined the effects of varying gravitational torques at the elbow and shoulder on the accuracy of forearm alignment to earth-fixed axes. Adding a 450 g load to the forearm to increase gravitational torques when the forearm is not vertical did not improve the accuracy of forearm alignment with the vertical. Furthermore, adding small, variably sized loads (between which the subjects could not distinguish at the perceptual level) to the forearm just proximal to the wrist produced similar errors in aligning the forearm with the vertical and horizontal. Forearm-positioning errors were not correlated with the size of the load, as would be expected if gravitational torques affected forearm-position sense. We conclude that gravitational torques exerted about the shoulder and elbow do not make significant contributions to sensing forearm-orientation relative to earth-fixed axes when the upper-limb segments are not constrained by external supports. PMID- 10382627 TI - Differences in functional magnetic resonance imaging of sensorimotor cortex during static and dynamic finger flexion. AB - Functional magnetic resonance imaging (fMRI) studies of the human motor system have commonly used movement paradigms which contain a dynamic component; however, the relationship between the fMRI signal for motor tasks with and without a dynamic component is not known. We have investigated the relationship between the fMRI signal during a static finger flexion task and during dynamic finger flexion at 1-3 Hz, each at two levels of force (5% and 10% of maximum voluntary contraction). A small fMRI response could be recorded from only a few subjects during the static tasks. In contrast, a substantial fMRI response occurred during dynamic tasks in all subjects at both levels of force. The fMRI response was not significantly correlated with force or movement rate during the dynamic tasks. It is concluded that the factors responsible for generating an fMRI response are fundamentally different during steady contractions compared to those involving a dynamic component, and that the fMRI signal may be more sensitive to changes in the pattern of neural activation rather than the ongoing firing rate or extent of activation. PMID- 10382628 TI - Inhibitory influences on receptive field size in the dorsal column nuclei. AB - Receptive fields (RFs) of neurons in the dorsal column nuclei (DCN) expand within minutes after the RFs are anesthetized via subcutaneous lidocaine injections (e.g., Pettit and Schwark). The mechanism of this rapid reorganization is of great interest. It has been proposed that such RF expansion results from a decline in inhibition within the DCN that unmasks previously ineffective synapses. To study the role of GABAergic inhibition in the DCN in controlling RF size, we applied by iontophoresis bicuculline methiodide to block gamma aminobutyric acidA (GABA(A)) receptors and 2-OH-saclofen to block GABA(B) receptors. Blockade of GABA(A) receptors resulted in RF expansions in 79% of the neurons, while blockade of GABA(B) receptors resulted in RF expansions in 53% of the neurons. The effectiveness of receptor blockade in producing RF expansion was not related to neuronal response characteristics. Glutamate application resulted in RF expansions in only 2 of 23 neurons tested, suggesting that RF expansion was not simply due to increased excitability. The modality and adaptation characteristics of the expanded portions of the RFs were similar to those of the original RF. The results of the present study suggest that GABAergic inhibition can play a role in controlling RF size in the DCN, and that both GABA(A) and GABA(B) receptors may be involved in this process. PMID- 10382629 TI - MR-eyetracker: a new method for eye movement recording in functional magnetic resonance imaging. AB - We present a method for recording saccadic and pursuit eye movements in the magnetic resonance tomograph designed for visual functional magnetic resonance imaging (fMRI) experiments. To reliably classify brain areas as pursuit or saccade related it is important to carefully measure the actual eye movements. For this purpose, infrared light, created outside the scanner by light-emitting diodes (LEDs), is guided via optic fibers into the head coil and onto the eye of the subject. Two additional fiber optical cables pick up the light reflected by the iris. The illuminating and detecting cables are mounted in a plastic eyepiece that is manually lowered to the level of the eye. By means of differential amplification, we obtain a signal that covaries with the horizontal position of the eye. Calibration of eye position within the scanner yields an estimate of eye position with a resolution of 0.2 degrees at a sampling rate of 1000 Hz. Experiments are presented that employ echoplanar imaging with 12 image planes through visual, parietal and frontal cortex while subjects performed saccadic and pursuit eye movements. The distribution of BOLD (blood oxygen level dependent) responses is shown to depend on the type of eye movement performed. Our method yields high temporal and spatial resolution of the horizontal component of eye movements during fMRI scanning. Since the signal is purely optical, there is no interaction between the eye movement signals and the echoplanar images. This reasonably priced eye tracker can be used to control eye position and monitor eye movements during fMRI. PMID- 10382631 TI - Update on clinical trials in the treatment of hepatitis B. AB - Chronic hepatitis B infection is a worldwide public health problem, which is particularly important in countries of Asia. Interferon has long been available for the treatment of patients with active replication (hepatitis B virus (HBV) e antigen and HBV-DNA positive) with evidence of chronic liver disease (elevated serum alanine aminotransferase and chronic hepatitis on liver biopsy). Doses of interferon of 10 MU, t.i.w. or 5 MU, q day for 16 weeks result in e antigen and HBV-DNA loss in approximately one-third of individuals who meet these treatment criteria. The major limitations of interferon are: (i) side effects of influenza like symptoms; (ii) need for parenteral administration; and (iii) concerns about safety in patients with hepatic decompensation. Nucleoside and nucleotide analogues have potent antiviral activity. The largest experience is with lamivudine (3-thiacytadine), a reverse transcriptase inhibitor that was recently approved by the USA Federal Drug Administration. At doses of 100 mg/day for 52 weeks, suppression of HBV replication is almost universal, with e antigen loss and improvement in histology being achieved in one-third and two-thirds of patients, respectively. The major advantages of lamivudine are: (i) good tolerability; (ii) oral route of administration; and (iii) safety in patients with hepatic decompensation. The major disadvantage is drug resistance, which is observed with increasing frequency following prolonged administration. New agents, such as adefovir dipivoxil, offer promise either alone or in combination with lamivudine in the treatment of individuals who are 'treatment naive' as well as in the treatment individuals who have developed lamivudine resistance. PMID- 10382630 TI - Natural history of chronic hepatitis B and C. AB - Hepatitis B virus (HBV) affects more than 300 million individuals worldwide and in the United States approximately 1.25 million individuals are chronic carriers of HBV. The risk of becoming a chronic hepatitis B virus surface antigen carrier is dependent upon the mode of acquisition of infection as well as the age of the individual at the time of infection. For those individuals with high levels of viral replication, chronic active hepatitis with progression to cirrhosis, liver failure and hepatocellular carcinoma (HCC) is common and liver transplantation is an excellent treatment option for patients with end-stage liver disease from HBV. Patients with chronic HBV infection should be screened periodically for hepatoma, although screening strategies have not been proven to prolong survival. Newer antiviral agents for the treatment of HBV are potent inhibitors of HBV-DNA and their long-term effect on the natural history of HBV is yet to be proven. The natural history of hepatitis C virus (HCV) infection is less well defined than that of chronic HBV. Certain patients who are chronic carriers of HCV may never develop extensive fibrosis, whereas others will progress to chronic active hepatitis with cirrhosis, HCC and end-stage liver disease. Factors that influence the progression of HCV are those related to the host, including the age at acquisition of infection, gender and immune status, and the disease process is accelerated in patients who consume regular amounts of alcohol. Hepatocellular carcinoma develops frequently in patients with HCV infection and its overall incidence is increasing due to this chronic viral disease. Patients with HCV cirrhosis should be screened regularly for hepatoma and liver transplantation is an effective treatment option for those with end-stage disease. The impact of antiviral therapy on the natural history of HCV is still to be determined and should be the focus of large clinical trials. PMID- 10382632 TI - Treatment strategies for chronic hepatitis C: update since the 1997 National Institutes of Health Consensus Development Conference. AB - The National Institutes of Health Consensus Development Conference on the management of hepatitis C, which took place in March 1997 and was published in September 1997, established guidelines for the diagnosis and management of chronic hepatitis C. The recommended treatment of chronic hepatitis C virus (HCV) infection is interferon alpha (or equivalent) 3 MIU three times per week for 12 months, in patients showing response to therapy after 3 months. Patients with the greatest risk for progression to cirrhosis (i.e. persistently elevated alanine aminotransferase levels, detectable serum HCV-RNA and liver biopsy showing portal or bridging fibrosis and at least moderate inflammation and necrosis) are recommended as candidates for therapy. The indication for therapy is less obvious in patients with milder histological changes, compensated cirrhosis and age less than 18 years or older than 60 years. Treatment is not indicated for patients with persistently normal aminotransferases or decompensated cirrhosis. This review outlines the background studies that led to the recommendations of the National Institutes of Health for the treatment of chronic hepatitis C and reviews newer evolving treatment strategies over the past year. In particular, the results of studies exploring treatment options for relapsers and non responders to prior interferon therapy and the reported results to date on the safety and efficacy of combination therapy with interferon plus ribavirin are highlighted. Although aggressive suppression of HCV-RNA with induction therapy (daily and/or higher doses) or long-acting pegylated interferon preparations may improve the current results of therapy, few data are yet available. Finally, the treatment of chronic hepatitis C with protease inhibitors holds promise but has yet to reach the stage of clinical trials. PMID- 10382633 TI - Antiviral therapy for hepatitis B and C in Asians. AB - The clinical features and treatment of chronic hepatitis C in Chinese patients are the same as in Caucasian patients except that 27% of Chinese chronic hepatitis C patients have hepatitis C virus (HCV) genotype 6a. In contrast, Chinese patients with chronic hepatitis B (CHB) differ from Caucasian patients because the Chinese patients are immunologically tolerant to hepatitis B virus (HBV), having acquired hepatitis B infection perinatally or in early childhood. In the treatment of CHB, the short-term aims of loss of hepatitis B virus early antigen (HBeAg) and HBV-DNA need to be reassessed. In 1296 Chinese CHB patients, 67.7% of those who developed complications of cirrhosis or hepatocellular carcinoma, were HBeAg-antibody positive. Longer follow up of patients is, therefore, required to assess the time efficacy of a treatment regimen. After long-term follow up (median 90 months) of 206 Chinese CHB patients treated with interferon alpha (IFN alpha) compared with 203 untreated subjects, IFN alpha conferred no benefit in cumulative HBeAg seroconversion or in HBV-DNA negativity as determined by polymerase chain reaction assays or in decreasing long-term complications of cirrhosis and hepatocellular carcinoma. Lamivudine is a novel nucleoside analogue. In a recent 1 year study in 358 Chinese CHB patients, lamivudine treatment was associated with substantial histological improvement (including a reduction in fibrosis), with HBV-DNA suppression and normalization of alanine aminotransferase levels. However, lamivudine may have to be given on a long-term basis, as withdrawal of lamivudine results in rebound of HBV-DNA to pretreatment levels. The long-term effects of lamivudine are currently being assessed. PMID- 10382634 TI - Advances in imaging techniques for the diagnosis of focal hepatic lesions. AB - In the past few years, tremendous advances have been made in the fields of ultrasound, computed tomography, magnetic resonance imaging and contrast agent development. The purpose of this article is to highlight the important developments in imaging techniques that can be used for detection and characterization of focal hepatic lesions. PMID- 10382635 TI - Major hepatic resection without blood transfusion: experience with total vascular exclusion. AB - Thirty consecutive, major liver resections performed with total vascular exclusion in both non-cirrhotic and cirrhotic patients were analysed retrospectively. The patients' ages ranged from 6 months to 80 years. Ten were Asians and five had cirrhosis associated with chronic hepatitis B or C. There was no perioperative death and the mean hospital stay was 6 days for adults and 9.2 days for children. The average vascular exclusion or warm ischaemia time was 25 min (range 10-55 min) and the average intraoperative blood volume given was 275 mL (range 0-3000 mL) packed red blood cells. Sixty per cent required no intraoperative blood transfusion. The mean total bilirubin and aspartate aminotransferase were 1.0 mg/dL (range 0.3-2.3 mg/dL) and 84 IU/L (range 14-306 IU/L) when measured prior to discharge at postoperative day 4-7. In our experience, total vascular exclusion is invaluable in major or difficult liver resections, especially lesions adjacent to the hepatic veins and vena cava. It is associated with a low blood transfusion requirement and a low incidence of complications. It further obviates the need for dissection of the porta hepatis and its associated risks. Total vascular exclusion time of 30 min appears to be well tolerated, even in patients with compensated cirrhosis. PMID- 10382636 TI - Role of alpha-fetoprotein in the diagnosis and management of hepatocellular carcinoma. AB - Thirty-five years after its first description, alpha-fetoprotein (AFP) remains the gold standard by which other markers are judged. Serum levels above the reference range of 10 ng/mL occur in approximately 75% of cases of hepatocellular carcinoma (HCC). In individual patients, the serum AFP level behaves as if it reflects tumour mass. However, the specificity of AFP is relatively low because moderately raised levels are also found in some patients with uncomplicated chronic liver disease. Recently, tumour-specific AFP assays have been developed. These are based on the carbohydrate side-chains on the AFP molecule which exhibit characteristic differences in AFP of different origins. Monitoring response to treatment may often be more effectively carried out by serial estimation of AFP than by conventional imaging techniques. The relative specificity of AFP for HCC has also been employed to detect circulating HCC cells and to target gene therapy. PMID- 10382637 TI - Resection versus transplantation for hepatocellular carcinoma. AB - Hepatocellular carcinoma is responsible for more than 1 million deaths per year worldwide and thus remains a challenging medical problem. It causes few or no symptoms and the tumour frequently reaches an enormous size by the time of diagnosis in countries where screening is seldom used. It is generally resistant to commercially available anti-neoplastic agents and radiation therapy. The principal treatment continues to be resection, either partial or complete, with liver transplantation. However, less than one-third of patients are surgical candidates for either resection or transplantation at the time of clinical presentation. This review will address the results observed following resection or transplantation for hepatocellular carcinoma. PMID- 10382638 TI - Patient selection and listing policies for liver transplantation. AB - Liver transplantation expanded rapidly during the first three decades of its application but has undergone slower growth over the past few years because of a limited supply of donor organs. The growing discrepancy between available donor organs and patients listed for transplantation has fuelled recent debate regarding patient selection criteria and listing policies for liver transplantation. Allocation and distribution regulations of the United Network for Organ Sharing have also recently undergone changes to balance the principles of utility (i.e. the overall benefits of liver transplantation to society) and justice (i.e. the needs of the individual patient). The ideal geographical area over which organs can be distributed to balance utility and justice remains uncertain, although there are pressures to widen sharing of donor organs. The sickest patient who has been waiting the longest still receive the next available donor organ, but listing policies for the sickest patients, those with fulminant hepatic failure or acute decompensation of chronic liver disease, were recently revised. A uniform minimal listing criteria that proposes listing patients when their estimated survival with liver disease is less than that expected after liver transplantation was recently proposed and has generally been accepted. Finally, cost-outcome analyses and the reality of managed care with the transfer of financial risk from insurers to providers is beginning to have an influence on patient selection criteria. PMID- 10382640 TI - Viral hepatitis and liver transplantation. AB - This article highlights the importance of hepatotropic viruses as pathogens in patients undergoing transplantation, their contribution to morbidity and mortality after transplantation and the approach to treatment of these pathogens when they cause disease. Although many advances have been made in the management of viral hepatitis in the transplant setting, there remain unanswered questions about the long-term natural history of the disease. Understanding of the pathogenesis of infection in the setting of transplantation is emerging slowly, but complete understanding requires further investigation. New approaches to treating disease in patients with either HBV or HCV infection are under development and will most likely focus on the use of combinations of antiviral and immunomodulatory agents. PMID- 10382639 TI - Does Asian race affect hepatitis B virus recurrence or survival following liver transplantation for hepatitis B cirrhosis? AB - To assess whether Asian race is an independent variable affecting survival and hepatitis B virus (HBV) recurrence after liver transplantation, the results of 27 consecutive liver transplants performed between June 1994 and April 1997 for HBV cirrhosis were analysed. In the group of 13 Asians, 38% had associated hepatocellular carcinoma and 62% had positive hepatitis B virus early antigen (HBeAg) or elevated HBV-DNA before transplant. Prophylactic hepatitis B immunoglobulin (HBIG) was administered perioperatively and long term at 4-6 weekly interval. Four patients with elevated HBV-DNA received lamivudine before transplantation. The 3 year actuarial patient survival rate was 100% in both Asian and non-Asian patients. Twenty-six patients remained seronegative for hepatitis B virus surface antigen after transplantation. The incidence of post transplant HBV recurrence was similar: 0% in Asians compared with 7% in non Asians. There was no recurrence in the group of 12 patients who were HBV-DNA or HBeAg negative pretransplant. PMID- 10382641 TI - Paediatric liver transplantation: indications, timing and medical complications. AB - Newer surgical techniques and immunosuppressive therapies have resulted in paediatric liver transplantation being available for most children with end-stage liver disease and has resulted in a greater than 80% 5-year survival rate. The most common indications for paediatric liver transplantation are biliary atresia (43%), metabolic disease (13%) and acute hepatic necrosis (11%). For approximately 75% of children with acute hepatic failure, the cause is unknown. Timing of liver transplantation not only affects survival rate, but may influence neurodevelopmental outcome. Fortunately, numerous types of donors, such as reduced-sized, living related or unrelated and blood-type mismatched, have reduced the mortality of children who are waiting for liver transplantation. However, the mortality and morbidity before and after liver transplantation remain high for children who have fulminant hepatic failure or are less than 5 months of age at the time of transplantation. The principle medical complications after liver transplantation are rejection and infection. Although use of newer immunosuppressive regimens has reduced the rate of rejection, Epstein-Barr virus infection with associated lymphoproliferative disorder remains the principle cause for morbidity and mortality after the initial 3 months post-liver transplant. PMID- 10382642 TI - Prevalence of Escherichia coli in apple cider manufactured in Connecticut. AB - Cider samples obtained from 11 cider mills operating in Connecticut during the 1997 to 1998 production season were tested for the presence of Escherichia coli. Cider production began in mid August and continued through March, with peak production in September and October. Of 314 cider samples tested, 11 (4%) were found to contain E. coli. Of the 11 mills, 6 (55%) tested positive for E. coli in the cider at least once during the production year. E. coli was first observed in cider samples produced in mid to late October and was not detected in samples made after January. A trend was observed for cider to decrease in acidity and increase in Brix (soluble sugars) throughout the production season. No correlation between pH and soluble sugars of cider and the presence of E. coli was detected. Eight mills used both dropped apples and tree-picked apples, whereas three mills used tree-picked apples only. The use of dropped apples in cider production began 5 weeks before the first detection of E. coli in cider. E. coli was isolated from cider samples produced using dropped apples and from samples produced using only tree-picked apples. No direct correlation between the use of dropped apples or tree-picked apples and the presence of E. coli in the cider was observed. An association between the time of apple harvest and the appearance of E. coli in cider was noted. For mills providing adequate records, all contaminated cider was produced from apples harvested between mid October and mid November. PMID- 10382643 TI - Fecal shedding and rumen growth of Escherichia coli O157:H7 in fasted calves. AB - Nine weaned calves aged from 8 to 12 weeks were fitted with rumen cannulas and were inoculated by cannula with 10(10) CFU of a five-strain mixture of nalidixic acid-resistant Escherichia coli O157:H7. Six calves were fasted for 48 h on days 15 and 16 and days 22 and 23 after inoculation. Samples of rumen contents and feces were obtained daily to enumerate E. coli O157:H7 populations and to determine rumen volatile fatty acid (VFA) concentrations and rumen pH. Fasting resulted in a marked decrease in rumen VFA concentrations from a mean of 135 mmol/liter before the fast to a mean of 35 mmol/liter during the second day of the fast. However, there was no correlation between daily VFA concentration and daily rumen or fecal numbers of E. coli O157:H7 in any of the calves. Fasting generally had no significant effect on the rumen or fecal numbers of E. coli O157:H7. The exception was a single fasted calf that experienced a 3-log(10) CFU/g increase in fecal shedding during and after the first fast. Despite the consistent changes in VFA concentrations in fasted calves, the fluctuations in rumen numbers of E. coli O157:H7 in the rumen of fasted calves were minimal. At the end of the experiment, E. coli O157:H7 was detected in either the rumen or omasum in two of three control calves at necropsy and in either the rumen or reticulum in five of six fasted calves. E. coli O157:H7 was detected in the colon in two of three control calves and in six of six fasted calves at necropsy. These results suggest that in cattle already shedding E. coli O157:H7, feed withdrawal and the associated changes in rumen pH and VFA concentrations have little effect on fecal shedding and rumen proliferation of E. coli O157:H7. PMID- 10382644 TI - Reduction of Escherichia coli O157:H7 and Salmonella typhimurium on beef carcass surfaces using acidified sodium chlorite. AB - The efficacy of a phosphoric acid-activated acidified sodium chloride (PASC) spray and a citric acid-activated acidified sodium chlorite (CASC) spray applied at room temperature (22.4 to 24.7 degrees C) in combination with a water wash was compared with that of a water wash only treatment for reduction of Escherichia coli O157:H7 and Salmonella Typhimurium inoculated onto various hot-boned individual beef carcass surface regions (inside round, outside round, brisket, flank, and clod). Initial counts of 5.5 and 5.4 log CFU/cm2 were obtained after inoculation with E. coli O157:H7 and Salmonella Typhimurium, respectively. Initial numbers for both pathogens were reduced by 3.8 to 3.9 log cycles by water wash followed by PASC spray and by 4.5 to 4.6 log cycles by water wash followed by CASC spray. The sprays consisted of applying 140 ml of the appropriate sanitizing solution for 10 s at 69 kPa. Corresponding reduction values obtained by water wash alone were 2.3 log. The performance of CASC appeared to be consistently better than that of PASC. In general, no effect of the carcass surface region was observed on the log reductions for either pathogen, except for the inside round, which consistently had lower reductions. Both PASC and CASC were capable of effectively reducing pathogens spread to areas beyond the initial contaminated area of the cuts to levels close to or below the counting method detection limit (0.5 log CFU/cm2). However, 30 to 50% of the carcasses treated by these antimicrobial solutions still yielded countable colonies. Results of this study indicate that acidified sodium chlorite sprays are effective for decontaminating beef carcass surfaces. PMID- 10382645 TI - Rapid growth of Salmonella enteritidis in egg white reconstituted from industrial egg white powder. AB - The aim of this study was to evaluate the consequences of the egg white-drying process on egg white ability to limit Salmonella Enteritidis growth in addition to the elucidation of the factors involved. We observed rapid growth of Salmonella Enteritidis inoculated in egg white reconstituted from industrial powder in comparison with that observed in liquid egg white collected in the laboratory: Salmonella cell counts rose from 10(3) to 10(8) cells/ml of egg white from powder during 24 h incubation at 30 degrees C. This rapid growth was observed in powder from all egg-breaking factories investigated, and it was comparable to that observed in optimum medium (tryptone soy broth). In view of the mechanism of egg white resistance and the major role played by iron availability and by ovotransferrin, we investigated several hypotheses to explain this rapid growth: iron provided during the drying process and/or denaturation of protein (especially ovotransferrin). The rapid growth observed in egg white reconstituted from powder was in relation to egg white protein denaturation and especially ovotransferrin denaturation during powder pasteurization that enhanced the availability of iron necessary for Salmonella growth. The major role played by ovotransferrin and iron deficiency on Salmonella growth in egg white was illustrated in this study. PMID- 10382646 TI - Use of mutant strain for evaluating processing strategies to inactivate Vibrio vulnificus in oysters. AB - Vibrio vulnificus is a ubiquitous marine bacterium frequently isolated from shellfish and associated with severe and often fatal disease in humans. Various control strategies to reduce the disease risk associated with V. vulnificus contamination in shellfish have been proposed. However, evaluating the efficacy of these control strategies is complicated because of the difficulty in distinguishing V. vulnificus from the high levels of background environmental Vibrio spp. The purpose of this research was to develop a model indicator V. vulnificus strain that could be readily differentiated from background microflora and used to facilitate the evaluation of processing efficacy. A spontaneous nalidixic acid-resistant strain of V. vulnificus (Vv-NA) was prepared from a wild type parent (Vv-WT) using selective plating techniques. Vv-NA was very similar to Vv-WT with respect to biochemical characteristics, appearance on selective plating media, detection limits using most probable number and polymerase chain reaction, and growth rate. In comparative freeze inactivation studies on pure cultures, Vv-WT and Vv-NA had similar freeze inactivation profiles at -20 degrees C (conventional freezing), at -85 degrees C (cold blast freezing), and in liquid nitrogen (cryogenic freezing). In oyster homogenates artificially inoculated with Vv-NA, the organism was inactivated 95 to 99% after freezing, irrespective of freezing temperature. Thermal inactivation comparisons of pure cultures of Vv-WT and Vv-NA using the capillary tube method revealed statistically significant differences in D values at 47 degrees C (2.2 versus 3.0 min, respectively) and 50 degrees C (0.83 versus 0.56 min, respectively), but nearly identical values at 52 degrees C (0.21 versus 0.22 min, respectively). However, these D values were notably higher than those reported by other investigators and hence provided a conservative means by which to evaluate thermal inactivation. In oyster homogenates seeded with Vv-NA, D values of 1.3+/-0.09 min and 0.41+/-0.01 min were obtained at 46 degrees C and 48 degrees C, respectively. This study demonstrated that Vv-NA is readily enumerated and could be used as a surrogate for evaluating the degree of V. vulnificus inactivation provided by freezing and thermal treatments of oyster homogenates. PMID- 10382648 TI - Comparison of three protocols for the isolation of Arcobacter from poultry. AB - The microaerophilic bacterium Arcobacter has received increasing attention in recent years regarding its presence in food products. There exist a limited number of methods for the detection of this microorganism, with currently available methods being cumbersome to perform, time consuming, and limited in specificity. The objective of this study was to develop a selective enrichment broth to isolate accurately three Arcobacter spp. from concentrated chicken microflora by comparing the efficacy of various selective and growth-promoting additives in order. This newly developed enrichment broth was incorporated into an isolation protocol using a previously developed plating medium, and this new protocol was compared with two existing methods for the isolation of Arcobacter from poultry. Method 1 consisted of enrichment in Ellinghausen-McCullough-Johnson Harris Polysorbate 80 broth followed by plating on cefoperazone-vancomycin amphotericin B medium. Method 2 consisted of enrichment in Arcobacter selective broth and plating onto Arcobacter selective medium. Method 3 (the JM method), used a newly developed enrichment broth followed by plating on a previously described JM agar. The JM method isolated Arcobacter strains in 42 out of 50 broiler chicken samples, while methods 1 and 2 detected the organism in only 24 and 15 out of 50 samples, respectively. PMID- 10382647 TI - Defining the growth/no-growth interface for Listeria monocytogenes in Mexican style cheese based on salt, pH, and moisture content. AB - The objective of this study was to define combinations of pH, salt, and moisture that produce growth, stasis, or inactivation of Listeria monocytogenes in Mexican style cheese. A soft, directly acidified, rennet-coagulated, fresh cheese similar to Mexican-style cheese was produced. The cheese was subsequently altered in composition as required by the experimental protocol. A factorial design with four moisture contents (42, 50, 55, and 60%), four salt concentrations (2.0, 4.0, 6.0, and 8.0% wt/wt), six pH levels (5.0, 5.25, 5.50, 5.75, 6.0, and 6.5), and three replications was used. Observations of growth, stasis, or death were obtained for each combination after 21 and 42 days of incubation at 10 degrees C. Binary logistic regression was used to develop an equation to determine the probability of growth or no growth for any combination within the range of the data set. In addition, ordinal logistic regression was used to calculate proportional odds ratios for growth, stasis, and death for each treatment combination. Ordinal logistic regression was also used to develop equations to determine the probability of growth, stasis, and death for formulations within the range of the data set. Models were validated with independently produced data. Of 60 samples formulated to have a 5% probability of Listeria growth (pH, 5.0 to 6.0; brine concentration, 8.17 to 16.00%), none supported growth. Of 30 samples formulated to have 50% probability of growth using the binary model (pH, 5.50 to 6.50; brine concentration, 3.23 to 12.50%), 20 supported growth. Of 30 samples formulated to have a 50% probability of growth according to the ordinal model (pH, 5.50 to 6.50; brine concentration, 3.37 to 10.90%), 16 supported growth. These data indicate that the logistic regression models presented accurately predict the behavior of L. monocytogenes in Mexican-style cheese. PMID- 10382649 TI - Antimicrobial resistance of bacterial flora associated with bovine products in South Africa. AB - The administration of subtherapeutic doses of antibiotics to livestock introduces selective pressures that may lead to the emergence and dissemination of resistant bacteria. This study determined the antibiotic-resistance spectra of the microbial flora found on freshly slaughtered and retail beef and in unpasteurized and pasteurized packaged milk. Staphylococci, Enterobacteriaeae, and isolates from total aerobic plate counts were tested for resistance to vancomycin, streptomycin, methicillin, tetracycline, and gentamicin using the disc diffusion susceptibility test and resistance to penicillin was determined by using oxacillin. A larger proportion of resistance to most antibiotics, except for vancomycin, was displayed by isolates from abattoir samples. The incidence of multiple antibiotic resistance (MAR) pathogenic bacteria is also higher in the abattoir. Resistance genes lost because of lack of selective pressure or resistant flora being replaced by more sensitive flora during processing is the reason for the lower incidence of MAR pathogenic bacteria among retail samples. These resistant bacteria can be transferred to humans through the consumption of rare or raw beef and unpasteurized milk, thus rendering the resultant food related infections difficult to treat. The present findings clearly demonstrate that antibiotic-resistant bacteria in beef and milk pose a serious problem in South Africa. PMID- 10382650 TI - Effects of antioxidants and gamma irradiation on the shelf life of beef patties. AB - To improve the storage safety of two types of ground beef patty popular in Korea (general beef patties and bulgogi patties), we added various antioxidants (200 ppm; including butylated hydroxyanisole, ascorbyl palmitate, alpha-tocopherol, and beta-carotene) to typical formulations of patties, cooked the patties to 70 degrees C, and irradiated them at a dose of 1.5 or 3 kGy. During 30 d of storage at 5 degrees C, the number of aerobic bacteria and lactic acid bacteria were determined using total aerobic plate count and phenyl ethyl alcohol-sucrose agar, respectively. The concentration of thiobarbituric acid was also determined in each type of patty. No colonies were observed in patties irradiated at 3 kGy regardless of which antioxidant was added. In control patties and patties with butylated hydroxyanisole that were irradiated at a dose of 1.5 kGy, growth of microorganisms appeared to be more rapid than in patties with natural antioxidants. The microbiological safety of nonirradiated patties could not be ensured for a period of 20 d. Lipid oxidation was retarded in both types of patty when an antioxidant was added. Ascorbyl palmitate had the strongest antioxidant effect among the natural antioxidants. However, butylated hydroxyanisole was more effective than ascorbyl palmitate when used in an equal amount. PMID- 10382651 TI - Identification and control of processing variables that affect the quality and safety of fluid milk. AB - The objective of this study was to increase quality and safety of fluid milk by eliminating postpasteurization contamination as measured by extended shelf life. Milk shelf life was defined as the number of days for standard plate count to reach 20,000 CFU/ml in milk stored at 7 degrees C. Sequential analysis of the fluid milk processing system indicated filling machine and pasteurizer were significant sources of postpasteurization contamination. Aseptically sampled milk from the pasteurizer outlet indicated a maximum shelf life of more than 30 days could be achieved. The pasteurizer can be a source of contamination when inadequately cleaned or maintained. The filling machine was a significant source of contamination. Shelf life of milk in 236-ml containers was reduced 20 days compared with milk sampled before the filling machine. Carton-forming mandrels, filling heads, and airborne microorganisms were sources of contamination within the filling machine. Eliminating sources of postpasteurization contamination and proper cleaning followed by sanitizing with chlorine significantly increased milk shelf life in paperboard containers to 20.4 days from an initial shelf life of 9 days. Changing the sanitizing agent to peroxyacetic acid significantly increased milk shelf life to 33.9 days. Enclosing the filling chamber and adding sterile laminar flowing air significantly improved microbiological quality of air inside the chamber and reduced variance among milk shelf life samples. PMID- 10382652 TI - Evaluation of a modified enzymatic test for the detection of tetracyclines in milk. AB - The tetracycline galactosidase (TG) test, a new method for the detection of tetracycline residues in raw milk based on the inhibition of beta-galactosidase biosynthesis in Escherichia coli, was previously validated with spiked milk samples. It has now been applied to milk from cows treated with oxytetracycline. In view of the occurrence of false positives, related to highly elevated somatic cell counts (>10(6)/ml), the improved TG test was developed, in which a heating step (80 degrees C, 15 min) preceded the original TG test protocol. A good agreement with other assays (Delvotest SP, the Bacillus cereus microtiter test, the LacTek tetracycline milk screening test, the Charm HVS-8100 tetracycline test) as well as with high-pressure liquid chromatography was obtained. No false negatives were observed with reference to the established maximum residue limit for tetracyclines of 100 microg/kg milk. PMID- 10382653 TI - Identification of inadequately cleaned equipment used in a sheep carcass-breaking process. AB - Aeromonads deposited on pork during a carcass-breaking process were recovered on hydrophobic grid membrane filters placed on ampicillin-dextrin-ethanol agar. Isolates from 85 honey-yellow colonies from filters on that medium were Aeromonas hydrophila (95%) or Vibrio sp. (5%). Presumptive aeromonads, coliforms, and Escherichia coli in swab samples from product passing through a sheep carcass breaking process were enumerated by hydrophobic grid membrane filtration techniques with a detection level of 1 CFU/100 cm2. Total aerobic counts were determined by a spread plate procedure with a detection level of 1 CFU/cm2 The numbers of aerobes, coliforms, and E. coli in the product were apparently unaffected by the carcass-breaking process, although coliforms and E. coli appeared to be redistributed from shoulder to loin and leg portions. However, the numbers of aeromonads on product increased by about two orders of magnitude as a result of the process. Few bacteria were recovered from most cleaned, large items of equipment, but aerobes, coliforms, and aeromonads were recovered at log total numbers of 5.25, 3.96, and 3.26, respectively, from most of 25 samples from bars supporting a conveyor belt. Also, aerobes, coliforms, E. coli, and aeromonads were recovered from 25 supposedly cleaned steel mesh gloves at log total numbers of 10.14, 5.54, 4.73, and 8.30, respectively. Those findings indicate that inspection of cleaned equipment and microbiological sampling of only food contacting surfaces, as is the current practice at meat plants, cannot provide assurance that the cleaning of carcass-breaking equipment is adequate. Instead, enumeration of indicator organisms on product passing through a process seems to be required as well, with subsequent sampling of equipment to identify sources of contaminants if increases in numbers during processing are observed. For surety of adequate cleaning, enumeration of several types of indicator organism may be necessary, as increases during processing in the numbers of organisms that are present in relatively large numbers on product entering a process may be difficult to detect. PMID- 10382654 TI - Bacterial contamination of ready-to-eat foods and fresh products in retail shops and food factories. AB - Raw vegetables cut for salad, cooked salad, cooked rice, boiled noodles, bean curd, and cooked Japanese foods were purchased in 27 retail shops in Tokyo. Intact vegetables before being processed and ready-to-eat fresh salad products were obtained from two food factories located in the suburbs of Tokyo. Two hundred thirty-eight retail samples, 137 samples of intact vegetables, and 159 samples of fresh products were examined for aerobic plate count (APC), coliforms, Escherichia coli, Listeria spp., Staphylococcus aureus, and Bacillus cereus. The APC of retail foods were 2.1 to 5.7 log CFU/g, and the range for the coliforms was 0.1 to 2.3 log CFU/g. The APC and coliform values showed that the raw vegetables cut for salad were the most heavily contaminated among the six kinds of ready-to-eat foods examined. Although L. monocytogenes was not detected, two samples of raw vegetables and five kinds of cooked foods yielded Listeria spp. S. aureus was detected in one sample of Japanese cooked food. The APC of the intact vegetables were 2.9 to 7.3 log CFU/g upon arrival and 2.2 to 7.2 log CFU/g after 3 days storage at 10 degrees C. The APC of the fresh products were 3.4 to 7.6 log CFU/g upon arrival and 4.7 to 8.7 log CFU/g after 3 days storage at 10 degrees C. The isolation rates for coliforms were 6.1 to 50% for intact vegetables and 50 to 66.7% for fresh products. E. coli was detected only in the fresh products. B. cereus was isolated from 20.1% (17 of 81) of the intact vegetables and 9.2% (8 of 87) of the fresh products. PMID- 10382655 TI - Aflatoxin reduction in corn through field application of competitive fungi. AB - Soil in corn plots was inoculated with nonaflatoxigenic strains of Aspergillus flavus and A. parasiticus during crop years 1994 to 1997 to determine the effect of application of the nontoxigenic strains on preharvest aflatoxin contamination of corn. Corn plots in a separate part of the field were not inoculated and served as controls. Inoculation resulted in significant increases in the total A. flavus/parasiticus soil population in treated plots, and that population was dominated by the applied strain of A. parasiticus (NRRL 21369). In the years when weather conditions favored aflatoxin contamination (1996 and 1997), corn was predominately colonized by A. flavus as opposed to A. parasiticus. In 1996, colonization by wild-type A. flavus was significantly reduced in treated plots compared with control plots, but total A. flavus/parasiticus colonization was not different between the two groups. A change to a more aggressive strain of A. flavus (NRRL 21882) as part of the biocontrol inoculum in 1997 resulted in a significantly (P < 0.001) higher colonization of corn by the applied strain. Weather conditions did not favor aflatoxin contamination in 1994 and 1995. In 1996, the aflatoxin concentration in corn from treated plots averaged 24.0 ppb, a reduction of 87% compared with the aflatoxin in control plots that averaged 188.4 ppb. In 1997, aflatoxin was reduced by 66% in treated corn (29.8 ppb) compared with control corn (87.5 ppb). Together, the data indicated that although the applied strain of A. parasiticus dominated in the soil, the nonaflatoxigenic strains of A. flavus were more responsible for the observed reductions in aflatoxin contamination. Inclusion of a nonaflatoxigenic strain of A. parasiticus in a biological control formulation for aflatoxin contamination may not be as important for airborne crops, such as corn, as for soilborne crops, such as peanuts. PMID- 10382656 TI - Medium optimization for the production of antioxidants from Aspergillus candidus. AB - The objective of this study was to optimize the factors for the production of antioxidant from Aspergillus candidus CCRC 31543. Extracts of broth filtrate had higher antioxidant activity (inhibition of peroxidation [IP] >98%) when sucrose or lactose was used as a carbon source. Sucrose in the medium also resulted in a higher yield of extracts. Ethyl acetate extracts had the highest yield and antioxidant activity compared with the other two solvents. For the production of antioxidant, inorganic nitrogen sources were found to be more suitable than organic nitrogen sources, and ammonium sulfate was better than sodium nitrate. Yeast extract had a strong influence on the yield of antioxidant extracts. Both mycelium and broth filtrate of A. candidus CCRC 31543 showed similar antioxidant activity (IP = 95%), and they also had similar extraction yields. PMID- 10382657 TI - Detection and isolation of Salmonella from naturally contaminated alfalfa seeds following an outbreak investigation. AB - Naturally contaminated alfalfa seeds, epidemiologically linked to foodborne disease outbreaks in Oregon and British Columbia, were tested for the presence of Salmonella. Ten sample units from the suspected lot were sprouted and grown for 4 days. After enrichment of the grown sprouts, an enzyme immunoassay (EIA) and culture method (modified procedure of the Food and Drug Administration Bacteriological Analytical Manual) were used for the detection and isolation of Salmonella. Four of the 10 sample units were positive with the EIA; however, 5 of the 10 sample units were culture positive (four were positive for Salmonella serotype Newport and a fifth was positive for Salmonella serotype Albany and serotype Schwarzengrund). The positive alfalfa seed sample units were further tested after shredding, soaking, and washing before culturing. Results suggest that sprouting and shredding methods may yield greater detection and recovery rates of Salmonella, but more research with a larger sample size is warranted. PMID- 10382658 TI - Effectiveness of various disinfectants in the elimination of Yersinia enterocolitica on fresh lettuce. AB - The effectiveness of various disinfectants against two potentially pathogenic Yersinia enterocolitica strains (Y. enterocolitica W1024 O:9 [strain A] and Y. enterocolitica B O:5 Lis Xz [strain B]) on shredded lettuce was examined. Dip wash treatments using 25, 100, and 300 ppm of chlorine at 4 and 22 degrees C, 0.2% Orenco Peel 40, 0.1% Tergitol, 0.5% acetic acid, and 0.5% lactic acid at 22 degrees C were performed. Surfactants and organic acids were also tested in combination with 100 ppm of chlorine. Reductions of Y. enterocolitica counts with 100 ppm (2.68 log10 for strain A and 2.36 log10 for strain B at 22 degrees C) and 300 ppm of chlorine (3.15 log10 for strain A and 2.55 log10 for strain B at 4 degrees C) were observed after 10 min. Inhibitory effect of different chlorine solutions was not significantly (P < 0.05) influenced by temperature. Surfactants in combination with chlorine were more effective than surfactants alone. Treatment with 0.2% Orenco Peel 40 plus 100 ppm of chlorine resulted in reductions of 2.69 log10 CFU/g for strain A and 3.18 1og10 CFU/g for strain B at 10 min. Dip solutions containing 0.1% Tergitol plus 100 ppm of chlorine produced a significant reduction of 2.73 log10 CFU/g in strain A (P < 0.05). With the 0.5% lactic acid plus 100 ppm of chlorine combination, inactivation of Y. enterocolitica was >6 log10. The bactericidal effect of disinfectants was related to the concentration, exposure time, combination with chlorine (surfactants and organic acids), and susceptibility of each strain. Since the presence of pathogenic Y. enterocolitica on ready-to-use vegetables represents a health hazard, treatments as effective as 0.5% lactic acid plus 100 ppm of chlorine are recommended for washing of fresh lettuce. PMID- 10382659 TI - Acid adaptation of Listeria monocytogenes strains does not offer cross-protection against an activated lactoperoxidase system. AB - Listeria monocytogenes has been implicated in foodborne illness outbreaks involving several types of cheeses made from acidified milk. Acid shock response (ASR) and acid tolerance response (ATR) could be possible reasons for its survival. The ASR and ATR of three strains of L. monocytogenes (V7, V37, and CA) in skim milk acidified to pH 4.0 and 3.5 with lactic acid and held at 32 degrees C were studied. Studies were also done to determine if acid adaptation of the organism enhanced survival in the presence of an activated lactoperoxidase system. The cells were directly shocked at pH 4.0 and 3.5 in skim milk to study the ASR. To study the ATR, cells were initially adapted in skim milk at a mild pH of 5.5 for the equivalent of one generation before being shocked at pH 4.0 and 3.5 in skim milk. Cells adapted at pH 5.5 in tryptic soy broth without dextrose and nonadapted cells were challenged at pH 4.5 in skim milk with or without an activated lactoperoxidase system. In all cases, viability and pH were measured 24 or 48 h after challenge. In pH 4.0 skim milk, for all three strains, the adapted cell population survived better (0.5 to 1.0 log higher) than that of nonadapted cells for 24 h. In pH 3.5 skim milk, the acid-adapted populations of all three strains were 3 to 4 logs greater than those of nonadapted cells at 6 h. The acid adapted cells of all three strains had survival rates similar to those of the nonadapted cells at pH 4.5 both in the presence and absence of an activated lactoperoxidase system. It was also evident that these strains do not exhibit an adaptive ATR at pH 4.5, although they do at lower pH levels (pH 4.0 and 3.5). Survival due to the ATR was better seen at pH 3.5 than at pH 4.0. PMID- 10382661 TI - Irradiation of chewable tobacco mixes for improvement in microbiological quality. AB - Microbiological quality of chewable tobacco mixes traditionally known as "Gutkha" was studied. The microbiological analysis of 15 samples analyzed revealed high bacterial and fungal counts. The total viable counts were in the range of 1.8 x 10(4) to 7.2 x 10(4) CFU g(-1) and the yeast and mold count from 3.6 x 10(3) to 7.1 x 10(4) CFU g(-1). The proteolytic and lipolytic counts were 9 x 10(2) to 2.6 x 10(3) CFU g(-1) and 2.6 x 10(3) CFU g(-1), on an average, respectively. Lecithinase-positive Staphylococcus aureus was found in 2 of the 15 samples analyzed; the counts were up to 3.4 x 10(3) CFU g(-1). Coliform and Salmonella spp. were found to be absent. Aflatoxins B , B2, and G2 were found to be present in all the samples. These samples were exposed to gamma radiation (60Co) at 1-, 2 , 3-, 5-, 10-, and 25-kGy doses. The decrease in total viable count and fungal count was noticed with increase of radiation dose. The 3-kGy dose was observed to be the sterilization dose for Gutkha. At this dose no survival of organisms was noticed and no revival was observed during postirradiation storage at room temperature for 6 months. PMID- 10382660 TI - Assessment of the microbiological conditions of tails, tongues, and head meats at two beef-packing plants. AB - Newly skinned tails of beef carcasses at two packing plants were similarly contaminated with total aerobes and with coliforms that were largely Escherichia coli at log mean numbers about 3.5/cm2 and 4.5/100 cm2, respectively. The log mean numbers of aerobes and coliforms on the skinned tails after washing at plant A were, respectively, 1 and 2 log units less than the numbers on the newly skinned tails. At plant B, the log mean numbers of aerobes on skinned and on washed tails were similar while the log mean numbers of E. coli on washed tails were only about 1 log unit less than the numbers on skinned tails. Cooling of tails on racks in a chiller at plant B reduced the log mean numbers of E. coli by about 1 log unit but did not reduce the numbers of total aerobes. Tongues in the heads of carcasses at both plants were similarly contaminated with total aerobes and with coliforms that were largely E. coli at log mean numbers of about 4.5/cm2 and 4.5/100 cm2, respectively. The log mean numbers of aerobes on and the log total number of E. coli recovered from washed tongues were, respectively, about 2 and 4 log units less than for unwashed tongues at plant A and about 1 and 3 log units less than for unwashed tongues at plant B. The log mean numbers of aerobes and E. coli on washed cheeks and lips were both about 2 log units less than the numbers on unwashed tongues at both plants. With appropriate collection and washing procedures, the microbiological conditions of beef tails, tongues, and head meats can apparently be comparable to those of primal cuts and manufacturing beef at the times that the products are packed. PMID- 10382662 TI - An oligonucleotide-ligation assay for the differentiation between Cyclospora and Eimeria spp. polymerase chain reaction amplification products. AB - An oligonucleotide-ligation assay (OLA) was developed and compared to a restriction fragment length polymorphism (RFLP) test for distinguishing between 294-bp polymerase chain reaction (PCR) amplification products of the 18S rRNA gene from Cyclospora and Eimeria spp. The PCR/OLA correctly distinguished between three Cyclospora, three E. tenella, and one E. mitis strains and the ratio of positive to negative spectrophotometric absorbance (A490) values for each strain ranged from 4.086 to 15.280 (median 9.5). PCR/OLA provides a rapid, reliable, spectrophotometric alternative to PCR/RFLP. PMID- 10382664 TI - Limitations of molecular biological techniques for assessing the virological safety of foods. AB - Enteric viruses, including hepatitis A, Norwalk, and Snow Mountain viruses, Hawaii agent, and rotaviruses have been associated with outbreaks of foodborne illness. Classical culturing procedures are available for poliovirus; however, hepatitis A, Norwalk, and many of the other viruses and agents cannot be propagated in cell culture, therefore, molecular biological tools have emerged as a possible means to detect enteric viruses in foods and environmental samples. There are limitations however in the application of polymerase chain reaction and reverse transcription polymerase chain reaction that restrict their usefulness for measuring the virological safety of foods. The most serious limitation is that molecular techniques fail to discriminate between viable and inactivated viruses even though inactivated viruses pose no threat to the consumer and may be present at levels substantially higher than the virulent forms. Other disadvantages include a lack of assay sensitivity and specificity, high assay costs, and a level of technical expertise not available in most food-testing laboratories. Overall, scientific advances in the development of molecular biological tools have outpaced the demonstration of their validity in assessing the virological safety of foods. PMID- 10382665 TI - Structure of the integral membrane domain of the GLP1 receptor. AB - A three-dimensional (3D) model of the integral membrane domain of the GLP1 receptor, a member of the secretin receptor family of the G-protein-coupled receptor superfamily is proposed. The probable arrangement of the seven helices in this receptor was deduced from a detailed analysis of all the sequences in the secretin receptor family. The analysis includes: 1) identifying the transmembrane helices, 2) charge distribution analysis to estimate to which extent the transmembrane helices are buried, 3) Fourier transform analysis of different property profiles within the transmembrane helices to determine the orientation of exposed and buried faces of the helices, 4) alignment of sequences with those of the rhodopsin-like family using the novel "cold spot" method reported herein, 5) determination of lengths of transmembrane helices and their connecting loops and the constraints these impose on packing, tilting and organization, 6) incorporation of mutagenesis and ligand specificity data. We find that there is a close similarity between the structural properties of receptors of the secretin family and those of the rhodopsin-like family as typified by the frog rhodopsin structure recently solved by electron cryomicroscopy. PMID- 10382663 TI - Evaluation of a fluorodensitometric method for analysis of ergosterol as a fungal marker in compound feeds. AB - Ergosterol is the principal sterol of fungi and plays an essential role as a component of the cell membrane and other cell constituents. This molecule is considered a good marker of fungal contamination in foods and feeds. This paper reports a rapid and sensitive method to test ergosterol content in compound feeds based on fluorodensitometry after thin-layer chromatography (TLC) separation. This method involves a thermal treatment of TLC plates that leads to the formation of a highly fluorescent ergosterol derivative. Such a dosage allows ergosterol testing in any naturally contaminated samples (limit of detection: 1 ppm of ergosterol) and gives results in close agreement with high-pressure liquid chromatography determination. Moreover, values obtained on mixed feeds for animals at different steps of fungal contamination are linked to quantitative development of storage fungi, evaluated by mycological technique, reinforcing the interest of a rapid method for measuring this fungal marker. PMID- 10382666 TI - Recognizing very distant sequence relationships among proteins by family profile analysis. AB - Family profile analysis (FPA), described in this paper, compares all available homologous amino acid sequences of a target family with the profile of a probe family while conventional sequence profile analysis (Gribskov M, Luthy R, Eisenberg D. Meth Enzymol 1990;183:146-159) considers only a single target sequence in comparison with the probe family. The increased input of sequence information in FPA expands the range for sequence-based recognition of structural relationships. In the FPA algorithm, Zscores of each of the target sequences, obtained from a probe profile search over all known amino acid sequences, are averaged and then compared with the scores for sequences of 100 reference families in the same probe family search. The resulting F-Zscore of the target family, expressed in "effective standard deviations" of the mean Zscores of the reference families, with value above a threshold of 3.5 indicates a statistically significant evolutionary relationship between the target and probe families. The sensitivity of FPA to sequence information was tested with several protein families where distant relationships have been verified from known tertiary protein architectures, which included vitamin B6-dependent enzymes, (beta/alpha)8 barrel proteins, beta-trefoil proteins, and globins. In comparison to other methods, FPA proved to be significantly more sensitive, finding numerous new homologies. The FPA technique is not only useful to test a suspected relationship between probe and target families but also identifies possible target families in profile searches over all known primary structures. PMID- 10382667 TI - Recognition of a protein fold in the context of the Structural Classification of Proteins (SCOP) classification. AB - A computational method has been developed for the assignment of a protein sequence to a folding class in the Structural Classification of Proteins (SCOP). This method uses global descriptors of a primary protein sequence in terms of the physical, chemical, and structural properties of the constituent amino acids. Neural networks are utilized to combine these descriptors in a way to discriminate members of a given fold from members of all other folds. An extensive testing of the method has been performed to evaluate its prediction accuracy. The method is applicable for the fold assignment of any protein sequence with or without significant sequence homology to known proteins. A WWW page for predicting protein folds is available at URL http://cbcg.lbl.gov/. PMID- 10382668 TI - Estimating the total number of protein folds. AB - Many seemingly unrelated protein families share common folds. Theoretical models based on structure designability have suggested that a few folds should be very common while many others have low probability. In agreement with the predictions of these models, we show that the distribution of observed protein families over different folds can be modeled with a highly-stretched exponential. Our results suggest that there are approximately 4,000 possible folds, some so unlikely that only approximately 2,000 folds existing among naturally-occurring proteins. Due to the large number of extremely rare folds, constructing a comprehensive database of all existent folds would be difficult. Constructing a database of the most-likely folds representing the vast majority of protein families would be considerably easier. PMID- 10382670 TI - High-resolution structures of scytalone dehydratase-inhibitor complexes crystallized at physiological pH. AB - Scytalone dehydratase is a molecular target of inhibitor design efforts aimed at preventing the fungal disease caused by Magnaporthe grisea. A method for cocrystallization of enzyme with inhibitors at neutral pH has produced several crystal structures of enzyme-inhibitor complexes at resolutions ranging from 1.5 to 2.2 A. Four high resolution structures of different enzyme-inhibitor complexes are described. In contrast to the original X-ray structure of the enzyme, the four new structures have well-defined electron density for the loop region comprising residues 115-119 and a different conformation between residues 154 and 160. The structure of the enzyme complex with an aminoquinazoline inhibitor showed that the inhibitor is in a position to form a hydrogen bond with the amide of the Asn131 side chain and with two water molecules in a fashion similar to the salicylamide inhibitor in the original structure, thus confirming design principles. The aminoquinazoline structure also allows for a more confident assignment of donors and acceptors in the hydrogen bonding network. The structures of the enzyme complexes with two dichlorocyclopropane carboxamide inhibitors showed the two chlorine atoms nearly in plane with the amide side chain of Asn131. The positions of Phe53 and Phe158 are significantly altered in the new structures in comparison to the two structures obtained from crystals grown at acidic pH. The multiple structures help define the mobility of active site amino acids critical for catalysis and inhibitor binding. PMID- 10382669 TI - Electrostatic reversal of serine proteinase substrate specificity. AB - Mouse granzyme B is a member of the chymotrypsin family of serine proteinases that has an unusual preference for cleavage of substrates following aspartate residues. We show here that granzyme B can be redesigned by a single amino acid substitution in one wall of the specificity pocket, arginine-226 to glutamate, to hydrolyze preferentially thioester substrates following basic amino acids. Amide substrates, however, were not hydrolyzed by the variant granzyme B. These results show that residue 226 is a primary determinant of granzyme B specificity and imply that additional structural components are required for catalysis of amide bonds. Molecular modeling indicated subtle variation in glutamate-226 orientation depending upon the state of protonation of the gamma-carboxylate, which may account for the secondary specificity of this enzyme for substrates containing phenylalanine. This represents the first example of electrostatic reversal of serine proteinase substrate specificity and suggests that residue 226 is a primary substrate specificity determinant in the granzyme B lineage of serine proteinases. PMID- 10382671 TI - A fast algorithm for genome-wide analysis of proteins with repeated sequences. AB - We present a fast algorithm to search for repeating fragments within protein sequences. The technique is based on an extension of the Smith-Waterman algorithm that allows the calculation of sub-optimal alignments of a sequence against itself. We are able to estimate the statistical significance of all sub-optimal alignment scores. We also rapidly determine the length of the repeating fragment and the number of times it is found in a sequence. The technique is applied to sequences in the Swissprot database, and to 16 complete genomes. We find that eukaryotic proteins contain more internal repeats than those of prokaryotic and archael organisms. The finding that 18% of yeast sequences and 28% of the known human sequences contain detectable repeats emphasizes the importance of internal duplication in protein evolution. PMID- 10382672 TI - Correlation between knowledge-based and detailed atomic potentials: application to the unfolding of the GCN4 leucine zipper. AB - The relationship between the unfolding pseudo free energies of reduced and detailed atomic models of the GCN4 leucine zipper is examined. Starting from the native crystal structure, a large number of conformations ranging from folded to unfolded were generated by all-atom molecular dynamics unfolding simulations in an aqueous environment at elevated temperatures. For the detailed atomic model, the pseudo free energies are obtained by combining the CHARMM all-atom potential with a solvation component from the generalized Born, surface accessibility, GB/SA, model. Reduced model energies were evaluated using a knowledge-based potential. Both energies are highly correlated. In addition, both show a good correlation with the root mean square deviation, RMSD, of the backbone from native. These results suggest that knowledge-based potentials are capable of describing at least some of the properties of the folded as well as the unfolded states of proteins, even though they are derived from a database of native protein structures. Since only conformations generated from an unfolding simulation are used, we cannot assess whether these potentials can discriminate the native conformation from the manifold of alternative, low-energy misfolded states. Nevertheless, these results also have significant implications for the development of a methodology for multiscale modeling of proteins that combines reduced and detailed atomic models. PMID- 10382673 TI - Steered molecular dynamics simulations of force-induced protein domain unfolding. AB - Steered molecular dynamics (SMD), a computer simulation method for studying force induced reactions in biopolymers, has been applied to investigate the response of protein domains to stretching apart of their terminal ends. The simulations mimic atomic force microscopy and optical tweezer experiments, but proceed on much shorter time scales. The simulations on different domains for 0.6 nanosecond each reveal two types of protein responses: the first type, arising in certain beta sandwich domains, exhibits nanosecond unfolding only after a force above 1,500 pN is applied; the second type, arising in a wider class of protein domain structures, requires significantly weaker forces for nanosecond unfolding. In the first case, strong forces are needed to concertedly break a set of interstrand hydrogen bonds which protect the domains against unfolding through stretching; in the second case, stretching breaks backbone hydrogen bonds one by one, and does not require strong forces for this purpose. Stretching of beta-sandwich (immunoglobulin) domains has been investigated further revealing a specific relationship between response to mechanical strain and the architecture of beta sandwich domains. PMID- 10382674 TI - Tryptophan microstate reshuffling upon the binding of cyclosporin A to human cyclophilin A. AB - Human cyclophilin A (hCypA) contains one tryptophan residue at position 121 (Trp121). The fluorescence intensity of this single tryptophan residue doubles upon binding the clinically important immunosuppressant cyclosporin A (CsA). Trp121 is in close contact to the bound CsA and is well-conserved in almost all immunophilins. The enhancement of the fluorescence intensity upon binding CsA is investigated by steady-state and time-resolved fluorescence measurements. The crystal structures of hCypA and the complex hCypA-CsA are compared. Only Glu120 is strongly influenced by the binding of CsA. The distance between the indole ring and the carboxylate group doubles during complexation. The influence of Glu120 on the fluorescence properties of Trp121 was investigated by pH-titration, and by substituting glutamate into an aspartate and an alanine residue. The fluorescence measurements on the glutamate mutants reveal that the carboxylate group influences the fluorescence properties of Trp121 to a limited extent. The major effect of CsA binding, however, consists in a reshuffling of the populations of microconformations of Trp121 leading to a selective increase of the 1.5 ns lifetime component. This selection is also accompanied by a decreased polarity of the environment and an increase in the radiative rate constant. PMID- 10382675 TI - Identification of the ligand binding site in Fas (CD95) and analysis of Fas ligand interactions. AB - Fas (CD95), a member of the tumor necrosis factor receptor superfamily, and its ligand (FasL), a tumor necrosis factor-like protein, are intensely studied because their interaction on the cell surface is critical for the induction of programmed cell death (apoptosis) and the regulation of immune responses. The structure and specificity of the extracellular binding domains of Fas and its ligand were studied, in different laboratories, by combining molecular modeling, mutagenesis, and a variety of binding and functional experiments. Residues critical for the receptor-ligand interaction were identified and, in the absence of experimentally determined structures, binding sites and details of the Fas ligand interactions were predicted. These studies provide an instructive example for the close combination of prediction and experiment and illustrate how insights into the structure and binding characteristics of Fas and its ligand were gradually refined. Discussed methodological aspects are representative of structure-function studies on extracellular domains of other single-path transmembrane proteins. PMID- 10382676 TI - Bradykinin and pituitary-adrenocortical function in the rabbit: in vitro and in vivo studies. AB - Bradykinin (BK) is a 9-amino acid peptide, which has been found to affect adrenocortical secretion in the calf and rat. We investigated the in vitro and in vivo effects of BK and its receptor antagonist [D-Arg, (Hyp3,D-Phe7)]-BK (BK-A) on pituitary-adrenocortical function in the rabbit. BK and BK-A raised basal release of aldosterone, but not of corticosterone by dispersed zona glomeralosa and zona fasciculata-reficularis cells, respectively. Both peptides did not affect ACTH-stimulated aldosterone secretion. Conversely, BK concentration dependently decreased ACTH-stimulated corticosterone production, and BK-A annulled this effect. The bolus intravenous injection of BK did not alter plasma ACTH concentration. However, BK lowered the blood concentration of both aldosterone and corticosterone, as well as the overall production of the two hormones over a period of 90 min after its administration. The simultaneous injection of BK-A blocked these effects of BK. BK-A alone did not evoke any sizeable change in blood hormonal levels. Collectively, these findings allow us to conclude that in rabbits (i) exogenous BK depresses adrenocortical secretion, through a receptor-mediated mechanism, which does not involve the inhibition of pituitary ACTH release-, and (ii) endogenous BK-like peptides do not play a relevant role in the functional regulation of the pituitary-adrenal axis, at least under basal conditions. PMID- 10382677 TI - Effects of physical exercise in pre-and postmenopausal women on lipid peroxidation and antioxidant systems. AB - The effects of moderate physical activity were studied in 19 postmenopausal and 25 premenopausal women on the enzymatic (superoxide dismutase and catalase) and non-enzymatic (vitamins A and E) antioxidant systems and on the processes of lipid peroxidation (malondialdehyde), and to see whether there is any variation in oxidative stress indicators between the postmenopausal and non-menopausal women. The subjects were also studied anthropometrically. At the end of a training period, there was a decrease in body fat percentage and in various skin folds. Neither group presented a rise in lipid peroxidation levels, although there was a significant (p<0.05) rise in superoxide dismutase activity in the postmenopausal women. PMID- 10382678 TI - Treatment with the NO-synthase inhibitor, methylene blue, moderates the decrease in serum leptin concentration in streptozotocin-induced diabetes. AB - It was previously reported that serum leptin concentrations were decreased in rats with streptozotocin-induced diabetes. Also, simultaneous nitric oxide (NO) synthase inhibitor treatment is known to partially attenuate streptozotocin induced diabetes development. The aim of our study was to investigate the influence of the NO-synthase inhibitor, methylene blue, on serum leptin concentration and diabetes development. Body weight, blood glucose, glycated hemoglobin and leptin concentration were measured in a control group, diabetic (streptozotocin 70 mg/kg i.p.) group, methylene blue (40 mg/kg in the food) treated group and a diabetic group treated with methylene blue. After six weeks of experiments, blood glucose and glycated hemoglobin increased significantly in the diabetic group vs controls (27.31 vs 5.49 mmol.l(-1), 14.11 vs 6.79%, respectively) and this increase was partially attenuated by simultaneous methylene blue treatment (16.8 vs 27.31 mmol/l, p < 0.05). Body weight and serum leptin fell in diabetic rats vs controls (248.9 vs 342.8 g, 0.57 vs 3.46 ng.ml( 1)). Treatment with methylene blue significantly suppressed the drop of body weight and the increase in blood glucose and glycated hemoglobin concentrations in the diabetic group. The decrease of serum leptin levels was significantly inhibited by methylene blue in the first experiment (1.1 vs 0.57 ng. ml(-1), p < 0.05); the same trend was found in a second experiment but the differences did not reach statistical significance. We conclude that the drop of serum leptin levels in diabetic rats is probably mainly due to streptozotocin-induced insulin deficiency, which is partially attenuated by NO-synthase inhibitor treatment. PMID- 10382679 TI - Characterization of an ACTH-induced chloride current in rat adrenal zona glomerulosa cells. AB - Adrenocorticotropic hormone (ACTH) is one of the principal activator of aldosterone secretion in rat zona glomerulosa cells, but its action on chloride currents is not well established. Here, we demonstrate that the hormone provoked a transient increase in a chloride current with a small unitary conductance estimated at 3.35 pS. Amplitude, as well as time-dependent increase of the ACTH induced chloride current was independent of the intracellular cAMP concentration. In contrary, its decrease was sensitive to alkaline phosphatase and PKA-inhibitor H-89, indicating that protein phosphorylation, at least in part via PKA, is involved in the decline of the current. PMID- 10382680 TI - The effects of growth hormone on corpus luteum of superovulated rats. AB - In general, growth hormone acts as a factor promoting cell proliferation in the positive direction and suppresses apoptosis. No report has described growth hormone (GH)-induced structural luteolysis. The present studies showed that GH induced structural luteolysis in rats after the induction of functional luteolysis by treatment with bromocriptine, and that apoptotic cells were present among luteal cells during structural luteolysis as shown by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling. Zymography showed that the activity of matrix metalloproteinase (MMP)-2 increased during GH induced structural luteolysis. The expression of c-myc protein of luteal cells was significantly decreased, but proliferating cell nuclear antigens (PCNA) were conversely increased during structural luteolysis, as shown by Western blot analysis. We propose that an excessive increase in PCNA and a marked decrease in c-myc protein of luteal cells lead to a disorder in the signals concerned with DNA synthesis, causing mitotic catastrophe and inducing apoptosis in luteal cells, and that structural luteolysis may be triggered. GH-induced apoptosis in structural luteolysis therefore highly depends on the cell cycle. There are thought to be two mechanisms of GH-induced structural luteolysis. One is apoptosis, and the other is destruction of extracellular matrix by MMP. PMID- 10382681 TI - Clinical characteristics of acromegaly in Hong Kong. AB - This study analyses the clinical characteristics of acromegalic patients in Hong Kong. All patients with acromegaly under follow up in Prince of Wales Hospital, Hong Kong between January 1984 and December 1992 were reviewed retrospectively. Detailed hospital notes were available for review in 28 out of 34. Of the 28 patients with full records available, 27 were Chinese and 1 was Nepalese. There were 8 (28.6%) males and 20 (71.4%) females. The mean age (+/- SD) at presentation was 51.2+/-16.8 years (range: 28 to 84 years) (male, 49.9+/-13.9 years [range: 28-66]; female, 51.7+/-18.1 years [range: 31-84]; p-value: NS). The commonest mode of presentation (n=22, 78.6%) was clinical suspicion by medical staff during consultation for other conditions, acromegaly being later confirmed. The estimated duration of symptoms, before diagnosis, was 14 years (range: 1 to 30 years). CT scan imaging of the pituitary gland showed that 12 patients (42.9%) had pituitary macro-adenomas (> or =1 cm), 3 (10.7%) had micro-adenomas (<1 cm), 6 (21.4%) had normal imaging, 1 (3.6%) had an empty sella and 6 (21.4%) had suspicious but inconclusive lesions in the pituitary gland. Surgery was offered as initial treatment to all patients. 4 to 6 weeks after surgery, if the maximal growth hormone response following glucose loading exceeded 10 microg/L, radiotherapy was offered. Of the 28 patients, 13 received surgery and radiotherapy, 2 surgery only, 4 radiotherapy only, 4 no treatment and 5 defaulted. At presentation, 50% had some abnormality of glucose tolerance. The mean early morning fasting baseline growth hormone was 52.8+/-37.0 microg/L (mean +/- SD, median: 48.1 microg/L) and the maximal growth hormone response during an extended oral glucose tolerance test was 63.2+/-34.9 microg/L (median: 61.3 microg/L). Forty five percents of patients had a maximal growth hormone response exceeding 60 microg/L. Of the 19 patients who underwent surgery and/or radiotherapy, 15 had their pituitary function reassessed 6 months after intervention. Their early morning fasting growth hormone and maximal growth hormone response in an extended oral glucose tolerance test were 21.3+/-25.8 and 35.4+/-37.5 microg/L, respectively. In conclusion, acromegaly in Hong Kong has an estimated annual incidence of 3.8 per million. There is a female preponderance, tendency to late presentation (>10 years) and low number of large tumors. Up to 80% were referred following observer suspicion. PMID- 10382682 TI - Prolactin secretion is increased in patients with multiple sclerosis. AB - Before the onset and during experimental allergic encephalomyelitis (EAE), the animal counterpart of multiple sclerosis (MS), prolactin levels were found to be elevated and bromocriptine was found to attenuate the attacks. This study was designed to determine whether patients with MS show evidence of hyperprolactinemia. Twelve patients with MS and twelve healthy controls were studied at baseline and with TRH stimulation, a provocative test for prolactin secretion. Compared to matched controls, patients with MS had slightly but significantly higher prolactin levels at baseline (10.2+/-1.6 vs 6.4 4+/-0.57 ng/ml, P=0.042), however, values were within the normal range. The prolactin levels post TRH were significantly higher in patients with MS: peak prolactin level was higher in patients than controls (57.08+/-6.144 vs 32.94+/-4.92 ng/ml, P=0.006). The area under the curve of prolactin was also higher in patients than in controls (3421.87+/-394.53 vs 2317.62+/-257.22 ng/ml, P=0.030). These findings are compatible with data from studies of experimental animals with MS and suggest that prolactin may play a role in the immunology of MS. PMID- 10382683 TI - Partial characterization of guanylyl cyclase activity in calf thyroid. AB - The aim of the present study was to perform the partial characterization of the enzyme guanylyl cyclase (GC) in bovine thyroid. The results obtained showed the presence of two types of GC: one is soluble and comprises around 79% of total activity, while the other is particulate. Treatment with 1% Triton X-100 increased both activities. When the kinetics of the enzyme was analyzed, using the complex MnGTP as a substrate, the results showed a Michaelis type kinetics for the soluble enzyme, with a Km of 0.037 mM, whereas the particulate GC showed a positive allosteric behavior with a S0.5 of 0.214 mM and a Hill coefficient of 1.9, indicating that the enzyme has at least two binding sites for the substrate. When the influence of different Mn2+ concentrations was studied, a positive allosteric behavior for the soluble GC was found, with a S0.5 of 1.2 mM and a Hill coefficient of 2.2. The kinetics of the particulate enzyme under similar conditions was of Michaelis type, with a Km of 0.752 mM. Although the enzyme is highly dependent on Mn2+, it was of interest to investigate the possible effects of other divalent cations, such as Ca2+ and Mg2+. The replacement of Mn2+ for Mg2+ caused a complete disappearance of the particulate enzyme activity, while the soluble activity decreased by 85%. Addition of Ca2+ had no effect on either GC. However, with suboptimum. concentrations of Mn2+, high Ca2+ concentration caused an increase in soluble activity, but it comprised only 20% of maximum activity with optimum Mn2+ concentrations. With the particulate enzyme a slight but significant inhibition was observed. PMID- 10382684 TI - Solitary nodular disease and multinodular goiter: a retrospective study on suppressive versus replacement levothyroxine therapy. AB - The aim of the present study was to ascertain whether patients affected by solitary nodular disease of the thyroid or multinodular goiter had a different clinical outcome when treated with suppressive levo-thyroxine (L-T4) therapy rather than replacement L-T4 therapy. We evaluated, by a retrospective analysis, 36 patients who had received TSH-suppressive L-T4 therapy according to TSH value and 55 who had received replacement L-T4 therapy. Fine needle aspiration cytology and thyroid scan after 131I were evaluated before L-T4 administration, while echographic monitoring of number and dimensions of nodules was recorded prior to and during L-T4 treatment. No difference in duration of L-T4 treatment (about 3 years) was registered between the TSH-suppressive therapy group and replacement therapy group. L-T4 administration in a TSH-suppressive or replacement manner did not induce a numerical or volumetric significant decrease of the main nodule or of the total nodule volume. Our data show that chronic TSH-suppressive therapy does not seem to be better than replacement therapy. Moreover, TSH-suppressive therapy presented a higher risk of adverse events than replacement therapy, thus requiring a more careful check with a higher cost of care. PMID- 10382685 TI - Paying respect to organs. PMID- 10382686 TI - Continuous positive airway pressure for treatment of sleep apnoea. PMID- 10382687 TI - Diagnosis and management of Cushing's syndrome. PMID- 10382688 TI - Haemochromatosis and control of intestinal iron absorption. PMID- 10382689 TI - Sensorimotor coordination in cerebral palsy. PMID- 10382690 TI - Incontinence after radical prostatectomy. PMID- 10382691 TI - Modern medicine: the state we're in. PMID- 10382692 TI - Diagnosis, prediction, and natural course of HIV-1 protease-inhibitor-associated lipodystrophy, hyperlipidaemia, and diabetes mellitus: a cohort study. AB - BACKGROUND: The prevalence and severity of lipodystrophy syndrome with long-term therapy for HIV-1 infection that includes a protease inhibitor is unknown. We studied the natural course of the syndrome to develop diagnostic criteria and identifying markers that predict its severity. METHODS: We assessed 113 patients who were receiving HIV-1 protease inhibitors (mean 21 months) and 45 HIV-1 infected patients (28 with follow-up) never treated with a protease inhibitor. Lipodystrophy was assessed by questionnaire (including patients' rating of severity), physical examination, and dual-energy x-ray absorptiometry. Body composition and fasting lipid and glycaemic variables were compared with data obtained 8 months previously. Oral glucose tolerance was investigated. FINDINGS: There was 98% concordance between patients' reports of the presence or absence of lipodystrophy (reported by 83% of protease-inhibitor recipients and 4% of treatment-naive patients; p=0.0001) and physical examination. Patients' ratings of lipodystrophy were significantly associated with declining total body fat (p=0.02). Lower body fat was independently associated with longer duration of protease-inhibitor therapy and lower bodyweight before therapy, and more severe lipodystrophy was associated with higher previous (p < 0.03) and current (p < or = 0.01) triglyceride and C-peptide concentrations, and less peripheral and greater central fat (p=0.005 and 0.09, respectively). Body fat declined a mean 1.2 kg over 8 months in protease-inhibitor recipients (p=0.05). The prevalence of hyperlipidaemia remained stable over time (74% of treated patients vs 28% of naive patients; p=0.0001). Impaired glucose tolerance occurred in 16% of protease inhibitor recipients and diabetes mellitus in 7%; in all but three patients these abnormalities were detected on 2 h post-glucose load values. INTERPRETATION: Diagnosis and rating severity of lipodystrophy is aided by the combination of physical examination, patient's rating, and measurement of body fat, fasting triglycerides, and C-peptide. Weight before therapy, fasting triglyceride, and C peptide concentrations early in therapy, and therapy duration seem to predict lipodystrophy severity. Lipodystrophy was common and progressive after almost 2 years of protease inhibitor therapy, but was not usually severe. Hyperlipidaemia and impaired glucose tolerance were also common. PMID- 10382693 TI - Comparison of therapeutic and subtherapeutic nasal continuous positive airway pressure for obstructive sleep apnoea: a randomised prospective parallel trial. AB - BACKGROUND: Nasal continuous positive airway pressure (NCPAP) is widely used as a treatment for obstructive sleep apnoea. However, to date there are no randomised controlled trials of this therapy against a well-matched control. We undertook a randomised prospective parallel trial of therapeutic NCPAP for obstructive sleep apnoea compared with a control group on subtherapeutic NCPAP. METHODS: Men with obstructive sleep apnoea, defined as an Epworth sleepiness score of 10 or more and ten or more dips per h of more than 4% SaO2 caused by obstructive sleep apnoea on overnight sleep study, were randomly assigned therapeutic NCPAP or subtherapeutic NCPAP (about 1 cm H2O) for 1 month. Primary outcomes were subjective sleepiness (Epworth sleepiness score), objective sleepiness (maintenance of wakefulness test), and SF-36 questionnaire measurements of self reported functioning and well-being. FINDINGS: 107 men entered the study: 53 received subtherapeutic NCPAP and 54 therapeutic NCPAP. Use of NCPAP by the two treatment groups was similar: 5.4 h (therapeutic) and 4.6 h (subtherapeutic) per night. Subtherapeutic NCPAP did not alter the overnight number of SaO2 dips per h compared with baseline, and thus acted as a control. Therapeutic NCPAP was superior to subtherapeutic NCPAP in all primary outcome measures. The Epworth score was decreased from a median of 15.5 to 7.0 on therapeutic NCPAP, and from 15.0 to 13.0 on subtherapeutic NCPAP (between treatments, p<0.0001). Mean maintenance-of-wakefulness time increased from 22.5 to 32.9 min on therapeutic NCPAP and, not significantly, from 20.0 to 23.5 min on subtherapeutic NCPAP (between treatments p<0.005). Effect sizes for SF-36 measures of energy and vitality were 1.68 (therapeutic) and 0.97 (subtherapeutic) NCPAP (between treatments p<0.0001). For mental summary score, the corresponding values were 1.02 and 0.4 (between treatments p=0.002). INTERPRETATION: Therapeutic NCPAP reduces excessive daytime sleepiness and improves self-reported health status compared with a subtherapeutic control. Compared with controls, the effects of therapeutic NCPAP are large and confirm previous uncontrolled clinical observations and the results of controlled trials that used an oral placebo. PMID- 10382694 TI - Prediction of benefit from carotid endarterectomy in individual patients: a risk modelling study. European Carotid Surgery Trialists' Collaborative Group. AB - BACKGROUND: Carotid endarterectomy lowers the risk of carotid territory ipsilateral ischaemic stroke, and is the treatment of choice, in patients with recently symptomatic 70-99% carotid stenosis. However, the 3-year risk of stroke on medical treatment alone is only about 20%. We investigated whether the efficacy of endarterectomy would be improved if patients with a high risk of stroke on medical treatment and a low risk of operative stroke or death could be identified. METHODS: We developed two prognostic models from data on patients with 0-69% carotid stenosis in the European Carotid Surgery Trial (ECST). The medical model predicted risk of ipsilateral carotid territory major ischaemic stroke (fatal or lasting longer than 7 days) on medical treatment and the surgical model predicted risk of major stroke and death within 30 days of endarterectomy. From these models we developed a prognostic score to identify patients with a high risk of stroke on medical treatment but a low operative risk. We validated the models and tested the scoring system on 990 ECST patients with 70-99% carotid stenosis assigned surgery (594) or medical treatment only (396). FINDINGS: When patients with 70-99% stenosis were stratified by the scoring system, which was based on seven independent prognostic factors, endarterectomy was beneficial in only 162 (16%) patients. The 5-year absolute risk of carotid territory ipsilateral major ischaemic stroke, operative major stroke, or death was lowered by 33% in the 16% of patients with a score of 4 or more (odds ratio 0.12 [95% CI 0.05-0.29], p<0.0001), but not in the other 828 (84%) patients (1.00 [0.65-1.54], p=0.7). INTERPRETATION: Many patients with recently symptomatic 70-99% carotid stenosis may not benefit from carotid endarterectomy. Validation of the predictive score is needed on external datasets, but risk-factor modelling could be useful to identify those patients in whom endarterectomy will be beneficial. PMID- 10382695 TI - Cancer incidence and mortality after radioiodine treatment for hyperthyroidism: a population-based cohort study. AB - BACKGROUND: Radioiodine is used increasingly as first-line treatment for hyperthyroidism, but concerns remain about subsequent risk of cancer, especially in those treated at a young age. We investigated cancer incidence and mortality in patients treated with radioiodine for hyperthyroidism. METHODS: We did a population-based study in 7417 patients treated in Birmingham, UK, between 1950 and 1991. We compared details of all cancer diagnoses and deaths in 1971-91 from the UK Office for National Statistics with data on cancer incidence and mortality for England and Wales specific for age, sex, and period. FINDINGS: During 72,073 person-years of follow-up, 634 cancer diagnoses were made, compared with an expected number of 761 (standardised incidence ratio [SIR] 0.83 [95% CI 0.77 0.90]). The relative risk of cancer mortality was also decreased (observed cancer deaths 448, expected 499; standardised mortality ratio [SMR] 0.90 [0.82-0.98]). Incidence of cancers of the pancreas, bronchus, trachea, bladder, and lymphatic and haemopoietic systems was lowered. Mortality from cancers at all these sites was also reduced but findings were significant only for bronchus and trachea. There were significant increases in incidence and mortality for cancers of the small bowel (SIR 4.81 [2.16-10.72], SMR 7.03 [3.16-15.66]) and thyroid (SIR 3.25 [1.69-6.25], SMR 2.78 [1.16-6.67]), although absolute risk of these cancers was small. INTERPRETATION: The decrease in overall cancer incidence and mortality in those treated for hyperthyroidism with radioiodine is reassuring. The absolute risk of cancers of the small bowel and thyroid remain low, but the increased relative risk shows the need for long-term vigilance in those receiving radioiodine. PMID- 10382696 TI - Signs and symptoms of Duchenne muscular dystrophy and Becker muscular dystrophy among carriers in The Netherlands: a cohort study. AB - BACKGROUND: Carriers of Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) may show muscle weakness or dilated cardiomyopathy. Studies focusing on skeletal-muscle involvement were done before DNA analysis was possible. We undertook a cross-sectional study in a population of definite carriers to estimate the proportion and to assess the clinical profile of carriers with symptoms. We also assessed a possible correlation between genotype and phenotype. METHODS: Carriers of DMD and BMD, aged 18-60 years, were traced through the files of the central register kept at the Department of Human Genetics in Leiden, Netherlands. For each carrier who agreed to participate a medical history was taken, and muscle-strength assessment by hand-held dynamometry and manual muscle testing and cardiological assessment were done. FINDINGS: 129 carriers of muscular dystrophy (85 DMD, 44 BMD) participated in the study. In 90 women from 52 (70%) families, 37 different mutations were found. 28 (22%) women had symptoms. 22 (17%) had muscle weakness, varying from mild to moderately severe. Muscle weakness was found in carriers of DMD and BMD, but dilated cardiomyopathy was found only in seven (8%) carriers of DMD, of whom one had concomitant muscle weakness. There was an unexpectedly high proportion of left-ventricle dilation (18%). No genotype-phenotype correlation was found. INTERPRETATION: Clinical manifestation of muscle weakness, dilated cardiomyopathy, or both can be found in about a fifth of carriers of DMD and BMD. If left-ventricle dilation is taken into account, the proportion of carriers with symptoms is even higher, amounting to 40%. PMID- 10382697 TI - Duodenal metal-transporter (DMT-1, NRAMP-2) expression in patients with hereditary haemochromatosis. AB - BACKGROUND: Although the gene for hereditary haemochromatosis has been cloned, the mechanism by which iron uptake is inappropriately increased in this disorder is unclear. Iron absorption is regulated by the duodenal metal transporter, DMT 1, also called NRAMP-2. We investigated the expression of NRAMP-2 in patients with hereditary haemochromatosis. METHODS: Duodenal biopsy samples were taken from 20 patients with haemochromatosis homozygous for the C282Y mutation and from ten controls. NRAMP-2 expression was assessed by northern blotting and competitive PCR. NRAMP-2 mRNA was sequenced in seven patients and two controls. FINDINGS: Duodenal NRAMP-2 mRNA concentrations were increased in patients as estimated by Northern blotting. Accordingly, competitive PCR showed significantly higher NRAMP-2 cDNA concentrations in patients than in controls (mean 3.43 [SD 0.61] vs 1.11 [0.74] log ng competitor x 10(4); p<0.001). No mutations were found within the NRAMP-2 mRNA. Duodenal NRAMP-2 mRNA expression was correlated with serum ferritin in controls (r=-0.94, p=0.001) but not in patients (r=-0.18, p=0.8). INTERPRETATION: Increased NRAMP-2 mRNA expression in duodenal mucosa of patients with hereditary haemochromatosis may promote duodenal iron uptake and lead to iron overload. PMID- 10382698 TI - Prevention before cure. PMID- 10382699 TI - Serial magnetic resonance imaging of cerebral atrophy in preclinical Alzheimer's disease. PMID- 10382700 TI - Radiation therapy and survival in choroid plexus carcinoma. PMID- 10382701 TI - Expression of leukaemia inhibitory factor in skin of patients with amyotrophic lateral sclerosis. PMID- 10382702 TI - Remarkable and persistent shrinkage of uterine leiomyoma associated with interferon alfa treatment for hepatitis. PMID- 10382703 TI - Importance of drug-device interaction in determining systemic effects of inhaled corticosteroids. PMID- 10382704 TI - Antibacterial activity of hyperforin from St John's wort, against multiresistant Staphylococcus aureus and gram-positive bacteria. PMID- 10382705 TI - Can we stop national immunisation days before global eradication of poliomyelitis? PMID- 10382706 TI - Lessons from the Nordic ICD pilot study. PMID- 10382707 TI - Call made for application of drug-donation guidelines. PMID- 10382708 TI - 54-week results on infliximab attract attention at EULAR. European League against Rheumatism. PMID- 10382709 TI - New molecular tool may lead to a new class of drugs. PMID- 10382710 TI - The National Institutes of Health flexes its muscle--but why now? PMID- 10382711 TI - Croatia and Bosnia: the imprints of war--I. Consequences. AB - As Serbia and Kosovo emerge from yet another European war, their people's health and the region's health care, scientific research, and medical education have been seriously damaged and disrupted. There are lessons to be learned from recent Balkan wars, lessons that might help doctors, international relief organisations, and governments to do better than they have done elsewhere during the long reconstruction period that will follow this recent savage conflict. An analysis of the medical legacies of war may also raise issues for doctors worldwide to consider as part of their role in a larger public-health community. For a week in May, 1999, I travelled to Croatia and the Croat-Muslim Federation of Bosnia Herzegovina to meet doctors working in peace but next to war. In the first part of this essay, I briefly survey some of the medical consequences of the Croatian and Bosnian conflicts. In the second part, to be published in the June 26 Issue, I consider plans for and limitations to restoration, and try to identify possible opportunities for prevention of the adverse health effects of war in a newly enlarged Europe. PMID- 10382712 TI - Visceral pain. AB - Visceral pain is the most common form of pain produced by disease and one of the most frequent reasons why patients seek medical attention. Yet much of what we know about the mechanisms of pain derives from experimental studies of somatic not visceral nociception. The conventional view is that visceral pain is simply a variant of somatic pain, a view based on the belief that a single neurological mechanism is responsible for all pain. However, the more we learn about the mechanisms of somatic and visceral pain, the more we realise that although these two processes have much in common, they also have important differences. Although visceral pain is an important part of the normal sensory repertoire of all human beings and a prominent symptom of many clinical conditions, not much clinical research has been done in this field and there are few clinical scientists with expertise in the management of visceral pain. Instead, visceral pain is usually treated by a range of specialists who take quite different approaches to the management of this type of pain. Thus, the management of visceral pain is frequently unsatisfactory. In this review, we consider visceral pain as a separate form of pain and examine its distinct sensory properties from a clinical perspective. We describe recent research findings that may change the way we think about visceral pain and, more importantly, may help develop new procedures for its management. PMID- 10382713 TI - The exceptional brain of Albert Einstein. PMID- 10382714 TI - Delay in diagnosis in breast cancer. PMID- 10382715 TI - Delay in diagnosis in breast cancer. PMID- 10382716 TI - Delay in diagnosis in breast cancer. PMID- 10382717 TI - Delay in diagnosis in breast cancer. PMID- 10382718 TI - Fetal growth velocity. PMID- 10382719 TI - Amantadine and pneumonia in elderly stroke patients. PMID- 10382720 TI - Amantadine and pneumonia in elderly stroke patients. PMID- 10382721 TI - Amantadine and pneumonia in elderly stroke patients. PMID- 10382722 TI - ABO blood group and improvement of preoperative fragment prothrombin 1,2 in orthopaedic surgery. PMID- 10382723 TI - Is change of the vessel wall a risk factor for venous thrombosis? PMID- 10382724 TI - Platelet counts and prognosis of pancreatic cancer. PMID- 10382725 TI - Corticosteroids in IgA nephropathy. PMID- 10382726 TI - Increased number of hip fractures. PMID- 10382727 TI - Increased number of hip fractures. PMID- 10382728 TI - Provision of primary total hip replacement surgery. PMID- 10382729 TI - Registration of new drugs in developing countries. PMID- 10382730 TI - Carbapenem-hydrolysing IMP-1 beta-lactamase in Klebsiella pneumoniae from Singapore. PMID- 10382731 TI - Sociological models and health care. PMID- 10382732 TI - In-vitro fertilisation in Israel. PMID- 10382733 TI - Dioxin. PMID- 10382734 TI - Stopping a clinical trial properly. PMID- 10382735 TI - The beautiful game. PMID- 10382736 TI - The Nobel chronicles. 1959: Severo Ochao (1905-1993) and Arthur Kornberg (born 1918). PMID- 10382737 TI - Influence of HLA-DR on the phenotype of CD4+ T lymphocytes specific for an epitope of the 16-kDa alpha-crystallin antigen of Mycobacterium tuberculosis. AB - T helper phenotype may be influenced by cytokine milieu, the differential expression of co-stimulatory molecules, antigen dose, and by differences in affinity at the TCR-peptide-MHC interface. We investigated the latter hypothesis by examining the response of six HLA-DR-restricted CD4+ T cell lines specific for the immunodominant and permissively recognized p91-110 epitope of the 16-kDa alpha-crystallin protein of Mycobacterium tuberculosis. Each line was generated from a sensitized HLA-DR-heterozygous donor and all proliferated when peptide was presented by autologous irradiated peripheral blood mononuclear cells. However, when HLA-DR-matched homozygous Epstein-Barr-virus-transformed B cell lines (L BCL) were used as peptide-presenting cells there was heterogeneity in the response. The most pronounced proliferative response, and the highest IFN-gamma secretion and cytolytic activity was stimulated by L-BCL expressing molecules (DRB1*0101, *1501 and *0401) with high affinity (IC50 < 10 microM) for the 16p91 110 peptide. By comparison, IL-4 secretion or a lower proliferative response could occur when peptide was presented by alleles of high, or of intermediate (10 microM < IC50 < 100 microM), affinity. These data support the hypothesis that the host MHC can influence CD4+ phenotype and have implications for subunit vaccination against tuberculosis. PMID- 10382738 TI - IL-12 directly up-regulates the expression of HLA class I, HLA class II and ICAM 1 on human melanoma cells: a mechanism for its antitumor activity? AB - IL-12 enhances cytolytic activity and proliferation of NK and T cells, and induces cytokines such as IFN-gamma. No direct effects on non-hematopoietic cells have been shown. This study investigates the effects of IL-12 on melanoma cells in vitro. We analyzed 15 melanoma cell cultures and 1 melanoma cell line. Out of 16 samples 13 expressed the beta chain of the IL-12 receptor (IL-12Rbeta). Preincubation with IL-12 increased the surface levels of human leukocyte antigen (HLA) class I, HLA class II and intercellular adhesion molecule (ICAM)-1 of those cultures with IL-12Rbeta expression. The effects of IL-12 on HLA class I could be blocked by an IL-12-neutralizing monoclonal antibody (mAb), but not by an mAb against IFN-gamma. Melanoma cells transduced with IL-12 expressed enhanced levels of HLA class I, HLA class II and ICAM-1 compared to controls. Co-incubation of the melanoma cells with allogeneic peripheral blood mononuclear cells (PBMC) resulted in enhanced proliferation and increased production of IL-2 and IFN-gamma after pretreatment with IL-12. IL-12 pretreatment increased the susceptibility of melanoma cells to lysis by prestimulated autologous PBMC. Since IL-12 induced immunocritical surface molecules on melanoma cells, it might be beneficial during immune interventions in melanoma patients. PMID- 10382739 TI - The mucosal adjuvant effects of cholera toxin and immune-stimulating complexes differ in their requirement for IL-12, indicating different pathways of action. AB - Adjuvants that can improve mucosal vaccine efficacy are much warranted. In this comparative study between cholera toxin (CT) and immune-stimulating complexes (ISCOM) we found that, contrary to CT, ovalbumin (OVA)-ISCOM were poor inducers of mucosal anti-OVA IgA responses, but induced similar or better systemic immunity following oral immunizations. The addition of CT to the oral OVA-ISCOM protocol did not stimulate local anti-OVA IgA immunity, nor did it change the quality or magnitude of the systemic responses. Both vectors recruited strong innate immunity, but only OVA-ISCOM could directly induce IL-12, demonstrable at the protein and mRNA levels. CT had no inhibitory effects on lipopolysaccharide/IFN-gamma-induced IL-12 mRNA expression or IL-12 production. Furthermore, adjuvanticity of CT was unaffected in IL-12-deficient mice, while OVA-ISCOM showed partly impaired adjuvant effects by the lack of IL-12. CT abrogated the induction of oral tolerance stimulated by antigen feeding in these mice. In addition, CT did not alter TGF-beta levels, suggesting that the immunomodulating effect of CT was independent of IL-12 as well as TGF-beta production. Taken together, these findings indicate that mucosal adjuvanticity of CT and ISCOM are differently dependent on IL-12, suggesting that separate and distinct antigen-processing pathways are involved. PMID- 10382741 TI - Cytotoxic T lymphocytes can induce a condemned state and synchronous post-mitotic apoptosis of daughter target cells. AB - We have used time-lapse video microscopy to study cytotoxic T lymphocyte (CTL) mediated apoptosis of LDb fibroblast target cells at different phases of the cell cycle. When aphidicolin-synchronized target cells were exposed to the CTL clone F5, apoptosis occurred with similar morphology during G1, S/G2 and M phase, showing that apoptosis and mitosis are not mutually exclusive cellular events. Interestingly, following normal mitosis of target cells that had been previously contacted by CTL, pairs of daughter cells would occasionally undergo apoptosis within minutes of each other. Such synchronous post-mitotic apoptosis was also observed when using mitotically unsynchronized target cells, and also when using d11S T cell hybridomas as alternative Fas- (CD95-) based effector cells, even if these effectors were physically washed away after an initial period of co incubation with the target cells. Our observations show that cytotoxic cells can induce a condemned state in pre-mitotic target cells, which can be inherited by both daughter cells, leading to their synchronous apoptosis after mitosis. PMID- 10382740 TI - TWEAK can induce cell death via endogenous TNF and TNF receptor 1. AB - TWEAK is a recently cloned novel member of the TNF ligand family. Here we show that soluble TWEAK is sufficient to induce apoptosis in Kym-1 cells within 18 h. TWEAK-induced apoptosis is indirect and is mediated by the interaction of endogenous TNF and TNF receptor (TNFR)1, as each TNFR1-Fc, neutralizing TNF specific antibodies and TNFR1-specific Fab fragments efficiently antagonize cell death induction. In addition to this indirect mode of action, co-stimulation of Kym-1 cells with TWEAK enhances TNFR1-mediated cell death induction. In contrast to TNF, TWEAK does only modestly activate NF-kappaB or c-jun N-terminal kinase (JNK) in Kym-1 cells. Although TWEAK binding to Kym-1 cells is easily detectable by flow cytometric analysis, we found neither evidence for expression of the recently identified TWEAK receptor Apo3/TRAMP/wsl/DR3/LARD, nor indications for direct interactions of TWEAK with TNFR. Together, these characteristics of TWEAK induced signaling in Kym-1 cells argue for the existence of an additional, still undefined non-death domain-containing TWEAK receptor in Kym-1 cells. PMID- 10382742 TI - Murine dendritic cells internalize Leishmania major promastigotes, produce IL-12 p40 and stimulate primary T cell proliferation in vitro. AB - Metacyclic Leishmania promastigotes (PM), transmitted by sand-fly bite, are likely to interact initially with cells of the dendritic cell (DC) lineage(s) in the epidermis or dermis. Epidermal Langerhans cells internalize L. major amastigotes (AM) and transport them to draining lymph nodes (Moll, H., Fuchs, H., Blank, C. and Rollinghoff, M., Eur. J. Immunol. 1993. 23: 1595) but little is known about the interaction of DC with PM. The present study demonstrates that DC are able to internalize PM and that the fate of the parasites within DC differs from that within macrophages (Mphi). DC took up small numbers of PM which did not differentiate into AM but appeared to be degraded; Mphi internalized large numbers of PM into parasitophorous vacuoles where they differentiated into AM. In response to direct stimulation with PM, DC from both C3H ("resistant" to L. major infection) and BALB/c ("susceptible") up-regulated production of IL-12 p40. In contrast, IL-12 production by Mphi was not detected. DC exposed to either metacyclic PM or PM culture supernatants were also able to stimulate proliferative responses in lymph node T cells from naive mice. These data indicate that DC have the capacity to promote protective Th1 immune responses in Leishmania infection and suggest that DC exposed to PM may be useful in immunotherapy and vaccination. PMID- 10382743 TI - Cellular, intracellular, and developmental expression patterns of murine SWAP-70. AB - SWAP-70 is part of a protein complex that catalyzes cell-free DNA recombination between immunoglobulin heavy chain gene switch region substrates. This report studies the expression pattern of SWAP-70 in mouse tissues, sorted cells, and cultured primary cells. SWAP-70 RNA is strongly increased upon switch-induction of spleen cells, and very weakly expressed in thymus and bone marrow. SWAP-70 protein is specifically expressed in B cells, and levels increase rapidly after stimulation. Tissue staining shows strong expression in germinal center B cells, while macrophages and T lymphocytes do not stain. SWAP-70 is not detected in early B cells in the bone marrow. Its expression during mouse ontogeny after birth correlates with the appearance of non-IgM isotypes. While SWAP-70 localizes to the cell nucleus in activated B cells, it is not tightly associated with the chromatin and is found in the cytoplasm as well. SWAP-70 expression is not increased by gamma or UV irradiation of spleen cells, nor does it depend on p53. These characteristics are consistent with the putative role of SWAP-70 in immunoglobulin class switching. PMID- 10382744 TI - Expression of CXCR4, the receptor for stromal cell-derived factor-1 on fetal and adult human lympho-hematopoietic progenitors. AB - Stromal cell-derived factor-1 (SDF-1) is a CXC chemokine produced by stromal cells that acts as a chemoattractant for human CD34+ progenitor cells. We investigated the expression of CXCR4, the receptor for SDF-1, on CD34+ cells from different hematopoietic sites and developmental stages. CXCR4 was detected by flow cytometry on 37 % of fetal bone marrow (BM) [gestation weeks (gw) 14-23] and 40% of adult BM CD34+ cells. Interestingly, in fetal liver CD34+ cells, CXCR4 was expressed at lower levels at later stages (9%, gw 20-23) compared to early stages of development (39%, gw 7.5-18), suggesting a development-related change in the migratory capacity of progenitors. CXCR4 was detected at similar levels on both phenotypically primitive and committed progenitors from fetal and adult sites. However, B cell lineage progenitor and precursor cells expressed CXCR4 at the highest density (80% of BM CD34+/CD10+ pro-B cells are CXCR4+). CXCR4 was also expressed in the fetal thymus in early T cell precursors and found to be down regulated during T cell maturation. Finally, we found that stem cell factor, alone or in combination with other cytokines, can up-modulate CXCR4 expression on CD34+ cells by three- to fourfold. In conclusion, our results suggest that CXCR4 may play an important role in the local and systemic trafficking of human CD34+ cells as well as in human B lymphopoiesis and that its expression can be modulated by cytokines. PMID- 10382745 TI - The lymphopenia mutation of the BB rat causes inappropriate apoptosis of mature thymocytes. AB - BB rats develop autoimmune diabetes mellitus at a high frequency. A key factor in the development of the disease is an autosomal recessive mutation determining peripheral T cell lymphocytopenia. Previous studies have suggested that the lymphopenia could be caused by increased cell death. Here we demonstrate that the lyp mutation dramatically reduces the in vitro lifespan of the TCRhi single positive thymocytes and peripheral T cells, without abolishing their capacity to proliferate. The reduced lifespan is due to an increased rate of apoptosis, and is detected in single-positive thymocytes displaying characteristics of cells which have undergone positive selection. The cell death defect does not affect the in vitro lifespan of peripheral B cells. Interestingly, stimulation can rescue peripheral lyp/lyp T cells from immediate cell death. We propose that the lymphopenia mutation prevents the accumulation of a normal T cell pool, including regulatory subsets, without preventing the activation and proliferation of reactive T cells, thereby creating conditions appropriate for the development of uncontrolled autoimmune responses. PMID- 10382746 TI - Tec kinase is involved in transcriptional regulation of IL-2 and IL-4 in the CD28 pathway. AB - The Tec protein tyrosine kinase (PTK) family includes Btk, Itk/Tsk/Emt, Tec, Rlk/Txk and Bmx, which are involved in signals mediated by various surface receptors. We have previously found (W.-C. Yang et al., J. Biol. Chem. 1999. 274: 607) that Tec is involved in T cell signaling in a way distinct from Itk. However, little is known about the role of Tec in regulation of cytokine expression in the CD28 pathway. Here, we show in heterologous COS-7 cells that co expression of Src family kinases such as Lck increases Tec activation or CD28 mediated Tec activation, whereas co-expression of kinase-dead Lck blocks Tec activation or CD28-mediated Tec activation. These data suggest that CD28 activates Tec via Src family PTK. As is the case for the IL-2 promoter, transcription of the IL-4 promoter is enhanced by overexpression of wild-type Tec but inhibited by overexpression of a kinase-dead version of Tec following CD28 activation. These results imply that Tec can modulate transcription of Th1 and Th2 cytokines in a kinase-dependent manner. Consistent with the hypothesis postulated above that Lck can regulate Tec activation, overexpression of kinase dead Lck can block Tec-induced cytokine expression following CD28 ligation. PMID- 10382747 TI - Therapeutic potential of TCR antagonists is determined by their ability to modulate a diverse repertoire of autoreactive T cells. AB - The use of altered peptide ligands (APL) with TCR antagonist properties holds promise as an antigen-specific therapy for autoimmune disorders. We are investigating the therapeutic potential of APL in experimental autoimmune encephalomyelitis (EAE) using the Ac1-9 peptide of myelin basic protein in H-2u mice. Encephalitogenic T cells recognize Ac1-9 using residues 3Gln and 6Pro as the major TCR contact sites. Use of position 6 APL is compromised by the heterogeneous nature of the Ac1-9-specific repertoire. Here we identify two position 3 APL that act as TCR antagonists on transgenic T cells expressing Ac1-9 specific TCR and that inhibit EAE in H-2u mice. However, the therapeutic capacity of these two APL correlated directly with the ability to maximally inhibit activation of a heterogeneous T cell pool. The implications of these findings for the requirements for EAE induction, the relative contribution of a given T cell subpopulation to pathology and the mechanism underlying EAE inhibition in this model are discussed. PMID- 10382749 TI - Functional and phenotypical characteristics of hepatic NK-like T cells in NK1.1 positive and -negative mouse strains. AB - We previously reported and partially characterized a unique monoclonal antibody (mAb), U5A2-13, which recognizes a T cell subset similar to NK1.1+ T cells, not only in NK1.1-positive mouse strains but also in NK1.1-negative strains. In NK1.1 positive C57BL/6 mice, U5A2-13+ TCRalphabeta+ cells produced abundant IL-4 as well as extremely high levels of IFN-gamma upon CD3 cross-linking, but this did not occur with U5A2-13- TCRalphabeta+ cells. In NK1.1-negative C3H/He mice, U5A2 13+ TCRalphabeta+ cells produced high levels of IL-4 and IFN-gamma upon CD3 cross linking, but this was not observed with U5A2-13- TCRalphabeta+ cells. To the best of our knowledge, this is the first direct evidence of the presence of NK-like T cells defined phenotypically by U5A2-13 mAb and functionally by IL-4/IFN-gamma production in NK1.1-negative mouse strains. We also demonstrated that U5A2-13- NK1.1+ T cells and U5A2-13+ NK1.1- T cells in C57BL/6 mice could produce both IL 4 and IFN-gamma. In addition, Vbeta8 or Vbeta7 usage by U5A2-13+ NK1.1- T cells was lower than that by U5A2-13+ NK1.1+ T cells, but remained higher than that by U5A2-13- NK1.1- T cells. Based on the present results, U5A2-13 mAb appears to be a valuable tool in the study of NK-like T cells. PMID- 10382748 TI - The N-terminal region of tapasin is required to stabilize the MHC class I loading complex. AB - Tapasin mediates the binding of MHC class I molecules to the transporter associated with antigen processing (TAP). Deletion mutants of tapasin were used to examine the effect of tapasin on interactions within the MHC class I complex. Binding to TAP is mediated by the C-terminal region of tapasin. Michaelis-Menten analysis of peptide transport shows that this interaction is sufficient to increase TAP levels without significantly affecting the intrinsic translocation rate. Weak interactions exist between MHC class I molecules and TAP in the absence of tapasin, and between free heavy chains and TAP-tapasin complexes in the absence of beta2-microglobulin. The N-terminal 50 residues of tapasin constitute the key element which converts the sum of these weak interactions into a stable complex. PMID- 10382750 TI - Effects of human immunodeficiency virus type 1 on CD4 lymphocyte subset activation. AB - The pathogenesis of the decline of CD4 lymphocyte counts accompanying the typical course of HIV-1 infection is not completely defined and might be related to a differential susceptibility of naive and memory cells to HIV-1 exposure. Here, we examined the effects induced by heat-inactivated HIV-1 virions on these lymphocyte populations. Exposure of CD45RA naive T cells to inactivated viral particles induced a marked decrease of both mitogenic responses and activation induced apoptosis. Conversely, the growth of CD45RO cells was less severely restrained. Analysis of intracellular levels of cell cycle regulatory proteins revealed an arrest at the G1/S restriction point of the naive but not memory subset. This effect was associated with alterations in phosphotyrosine profile and with a marked decrease of ERK and NJK kinase activation. Finally, up regulation of the cAMP-dependent protein kinase A (PKA) activity induced by mitogens was not affected by virus. Altogether, these findings show that interaction of HIV-1 with the T cell surface is sufficient to inhibit the proliferative response of the CD4CD45RA subset by disturbing proximal TCR signaling. This mechanism would affect renewal of naive lymphocytes, contributing in such a way to the impairment of T cell turnover during the course of HIV-1 infection. PMID- 10382751 TI - Pyrrolidine dithiocarbamate protects mice from lethal shock induced by LPS or TNF alpha. AB - Although important advances have been made in the development of antibiotics and medical intensive care technology in recent years, systemic response to infection remains a major health problem, with growing incidence and high mortality rates. Here we demonstrate the ability of the antioxidant agent pyrrolidine dithiocarbamate (PDTC) to inhibit the in vivo activation of NF-kappaB in lung and liver tissues, as well as the systemic release of TNF-alpha in lipopolysaccharide (LPS)-treated mice. The in vivo effect of PDTC on NF-kappaB activation in liver tissues involved the inhibition of both LPS-induced I kappaB-alpha degradation and the translocation of the p50 and p65 NF-kappaB subunits to the nucleus. In addition to protecting mice against lethal LPS doses, PDTC curtailed TNF-alpha induced lethal shock. This effect was observed even after LPS injection, and when PDTC was administered at a time when TNF-alpha was already at maximum levels in serum. PDTC-treated mice survived despite high IL-1beta and IL-6 levels, induction of VCAM-1 and ICAM-1 expression or leukocyte infiltration in tissues known to be associated with LPS-induced shock, indicating that PDTC does not act by modifying these responses. Taken together, these results indicate that PDTC interferes with the production as well as the action of TNF-alpha, and points to a possible approach toward the treatment of septic shock. PMID- 10382752 TI - Molecular mimicry between bacterial and self antigen in a patient with systemic lupus erythematosus. AB - The importance of microbial infection as a trigger for the induction of systemic lupus erythematosus is frequently debated. Clinical observations indicate that anti-viral and antibacterial responses are often accompanied by self reactivity, and anti-pneumococcal antibodies elicited in non-autoimmune individuals by pneumococcal vaccine express lupus-associated anti-DNA idiotypes. To explore the relationship between protective and pathogenic antibodies in humans, we have used the phage display immunoglobulin expression system to generate a combinatorial library from spleen cells of a lupus patient immunized with a polyvalent pneumococcal polysaccharide vaccine prior to splenectomy. From this library, monovalent antigen-binding fragments expressing the 3I Vkappa1-associated idiotype were isolated. This idiotype is expressed on up to 90% of anti-DNA antibodies in the serum of lupus patients and on anti-pneumococcal antibodies in the serum of non-autoimmune individuals. Eight 3I+ monovalent antigen-binding fragments reacting with pneumococcal polysaccharide, DNA or both were analyzed. Four of these fragments were cross-reactive with both foreign and self antigen, demonstrating that a high percentage of anti-bacterial antibodies produced in a patient with lupus bind double-stranded DNA. These studies provide support at the molecular level for a potential role of molecular mimicry in the generation of anti-DNA antibodies. In addition, this is, to our knowledge, the first panel of fully sequenced human anti-pneumococcal antibodies. PMID- 10382753 TI - Expression of the transcription factor GATA-3 is required for the development of the earliest T cell progenitors and correlates with stages of cellular proliferation in the thymus. AB - GATA-3 is a zinc-finger transcription factor that is essential for both early T cell development and Th2 cell differentiation. To quantify GATA-3 expression during T cell development in vivo in the mouse, the GATA-3 gene was targeted by insertion of a lacZ reporter by homologous recombination in embryonic stem (ES) cells. Although we could detect GATA-3+ cells throughout T cell development in the thymus, the proportions of GATA-3+ cells varied considerably between the distinct differentiation stages. The two periods of TCR alpha and beta gene recombination, which occur in quiescent or slowly dividing cells, were associated with low proportions of GATA-3+ cells. Conversely, the stage of rapidly proliferating cells, which insulates these two waves of TCR rearrangement, was characterized by a large proportion of GATA-3+ cells. In addition, we generated chimeric mice by injection of GATA-3-deficient, lacZ-expressing ES cells into wild-type blastocysts. In this in vivo competition analysis, no contribution of GATA-3-deficient cells to the T cell lineage was detected, not even in the earliest CD44+CD25- double-negative (CD4-CD8-) cell stage in the thymus. These results parallel data implicating other GATA family members as key regulators of proliferation and survival of early hematopoietic cells. We therefore propose that GATA-3 is required for the expansion of T cell progenitors, and for the control of subsequent proliferation steps, which alternate periods of TCR recombination in the thymus. PMID- 10382754 TI - Reassessment of the role of CD8+ T cells in the induction of allograft tolerance by donor-specific blood transfusion. AB - Donor-specific allograft tolerance can be induced in adult rats by pregraft donor specific blood transfusion (DST). We have previously shown that DST elicits in recipients the expansion of a donor-specific CD8+ T cell clone displaying the Vbeta18-Dbeta1-Jbeta2.7 TCR rearrangement, which rapidly infiltrates allografts after transplantation, suggesting a regulatory function for this clone in DST induced tolerance. In this study, recipients were pretreated before grafting, using an anti-CD8 monoclonal antibody to deplete CD8+ T cells. CD8 depletion before DST and transplantation abrogated allograft tolerance, and the CD8+ T cell clone was absent from allografts. These effects were not observed when CD8 depletion was performed after DST but before transplantation. We conclude that CD8+ T cells play a role in the induction of allograft tolerance by DST. PMID- 10382755 TI - Fractalkine and macrophage-derived chemokine: T cell-attracting chemokines expressed in T cell area dendritic cells. AB - Dendritic cells (DC) are a system of antigen-presenting cells specialized in interaction with T cells. Recently it has been reported that DC can produce CC (beta) chemokines that attract T cells. In this study we isolated mouse fractalkine and macrophage-derived chemokine (MDC) belonging to CX3C (delta) and CC chemokine families, respectively, from bone marrow-derived mature DC. While expression of fractalkine, which has so far been only examined in the brain and in vitro endothelial cells so far, was rather ubiquitous, MDC, which has been reported to be synthesized by macrophages and DC, was expressed specifically in the thymus and lymph node. This is the first report that indicates fractalkine expression by DC. Expression of fractalkine and MDC mRNA increased with maturation of DC during in vitro culture of bone marrow cells. Spleen- and epidermis-derived mature DC in culture also expressed these chemokines. Furthermore, their expression was detected selectively by Northern hybridization in CD11c+ B220- DC freshly purified from lymph nodes, and in large stellate cells in the lymph node T cell areas by in situ hybridization. Conditioned media of 293T cells transfected with these chemokine cDNA were chemotactic to Con A activated splenic T cells as well as the mouse T cell line EL4. In conclusion, while fractalkine and MDC belong to different families of chemokines, both may be involved in recruitment of T cells for interaction with mature DC in the immune response. PMID- 10382756 TI - The involvement of IL-12 in murine experimentally induced autoimmune thyroid disease. AB - Experimental autoimmune thyroid disease (EAT) can be induced experimentally in mice following immunization with mouse thyroglobulin (mTg) and the adjuvants lipopolysaccharide (LPS) or complete Freund's adjuvant (CFA). EAT can also be transferred to naive recipients by CD4+ T cells from mTg-primed mice. Here we demonstrate a role for IL-12 in the development of EAT by the ability of neutralizing antibody to IL-12 to reduce disease severity and by the lack of significant levels of thyroid infiltration in IL-12p40-deficient mice following immunization with mTg and CFA. A single injection of 300 ng IL-12 at the time of initial immunization with mTg and LPS was able to increase the degree of thyroid infiltration. These data are all consistent with EAT being a Th1-mediated disease. Conversely, however, administration of IL-12 over a prolonged period markedly inhibited the induction of EAT by mTg and CFA and, if given to recipients, inhibited the transfer of EAT by mTg-primed lymph node cells. The development of an autoantibody response to mTg was also inhibited when IL-12 was administered throughout the experimental period, suggesting that sustained exposure to IL-12 can be immunosuppressive. PMID- 10382757 TI - Broadly distributed T cell reactivity, with no immunodominant loci, to the pre erythrocytic antigen thrombospondin-related adhesive protein of Plasmodium falciparum in West Africans. AB - Protective immunity to malaria has been achieved in human volunteers utilizing the pre-erythrocytic Plasmodium falciparum antigen, the circumsporozoite protein (CS). However, T cell reactivity to CS is focused on several highly polymorphic T cell epitope regions, potentially limiting the efficacy of any vaccine to specific malaria strains. Another important pre-erythrocytic malaria antigen, the thrombospondin-related adhesive protein (TRAP), can induce protection in animal models of malaria, but knowledge of human T cell responses is limited to the identification of CD8 T cell epitopes, with no CD4 epitopes identified to date. This comprehensive study assessed reactivity to overlapping peptides spanning almost the whole of P. falciparum TRAP (PfTRAP), as well as peptides selected on the basis of HLA class II-binding motifs. A total of 50 naturally exposed Gambian adults were assessed to define 26 T cell epitopes in PfTRAP capable of inducing rapid IFN-gamma or IL-4 production, as assessed by enzyme-linked immunospot assays. In contrast to the CS protein, this reactivity was broadly distributed along the length of TRAP. Moreover, of the 26 epitopes identified, 10 were found to be conserved in West Africa. PMID- 10382758 TI - Structure-activity relationships within the N-terminal short consensus repeats (SCR) of human CR1 (C3b/C4b receptor, CD35): SCR 3 plays a critical role in inhibition of the classical and alternative pathways of complement activation. AB - Genes coding for between one and four short consensus repeats (SCR) of the N terminal region of human complement receptor 1 (CR1) were synthesized from oligonucleotides and those encoding SCR(1-2), SCR(1-3), SCR(1-4), SCR3 and SCR(3 4) were expressed as inclusion bodies in Escherichia coli. Following solubilization in urea, the proteins were partially purified and refolded and the activity of each protein was assessed in both classical and alternative pathway complement assays. All fragments showed a varying degree of activity with the general order being SCR(1-3) = SCR(1-4) > SCR(1-2). Addition of SCR3 to SCR(1-2) significantly improved potency, whereas the addition of SCR4 conferred no additional benefit. This observation, coupled with the ability of the single domain SCR3 to inhibit classical pathway mediated lysis with an IH50% (inhibition of hemolysis by 50%) of 4.8 microM, demonstrates that SCR3 provides key binding interactions with activated complement components. SCR(1-3) was able to inhibit both classical and alternative pathways of complement activation, showing that the N-terminal SCR of CR1 retain the ability to interact with C3b. Assays for CR1 like cofactor activity for factor I using C4b-like C4 or C3b-like C3 as substrates showed that SCR(1-3) possessed such cofactor activity and that C4b like C4 was a better substrate. When compared to full-length (30 SCR) soluble CR1 (sCR1), SCR(1-3) was significantly less potent in accord with a model involving multi-valent binding of C3b/C4b to CR1. PMID- 10382759 TI - Absolute requirement for the pre-T cell receptor alpha chain during NK1.1+ TCRalphabeta cell development. AB - Most natural killer T (NKT) cells express a highly skewed alphabeta TCR repertoire, consisting of an invariant V alpha14-J alpha281 chain paired preferentially with a polyclonal Vbeta8.2 chain. This repertoire is positively selected by the monomorphic CD1d molecule expressed on cells of hematopoietic origin. The origin of NKT cells and their lineage relationship to conventional T cells is controversial. We show here that the development of NKT cells is absolutely dependent on expression of the pre-TCRalpha chain, in marked contrast to conventional T cells which arise in significant numbers even in the absence of a functional pre-TCR. Distinct developmental requirements for pre-TCR expression in the NKT and T cell lineages may reflect differences in the ability of the TCRalphabeta to substitute functionally for the pre-TCR in immature precursor cells. PMID- 10382760 TI - Protection against aerosol Mycobacterium tuberculosis infection using Mycobacterium bovis Bacillus Calmette Guerin-infected dendritic cells. AB - In the lung, dendritic cells (DC) are key antigen-presenting cells capable of triggering specific cellular responses to inhaled pathogens, and thus, they may be important in the initiation of an early response to mycobacterial infections. The ability of DC to enhance antigen presentation to naive T cells within the lungs was characterized with respect to Mycobacterium bovis Bacillus Calmette Guerin (BCG) vaccination against M. tuberculosis infection. In vitro derived DC were infected with BCG, which induced their maturation, as shown by the increased expression of MHC class II antigens, CD80 and CD86 co-stimulatory molecules. The synthesis of mRNA for IL-1, IL-6, IL-12, IL-10 and IL-1 receptor antagonist was also enhanced. When administered intratracheally in mice, infected DC induced a potent T cell response and the production of IFN-gamma to mycobacterial antigens in the mediastinal lymph nodes, leading to a significant protection against aerosol M. tuberculosis infection. Intriguingly, although the vaccination schedule for BCG-infected DC was much shorter than subcutaneous BCG vaccination (7 days as compared to 100 days), both types of vaccination showed similar levels of protection. These data confirm that DC can be potent inducers of a cellular immune response against mycobacteria and support the concept of combining DC strategies with mycobacterial vaccines for protective immunity against tuberculosis. PMID- 10382761 TI - Fcgamma receptor-mediated inhibition of human B cell activation: the role of SHP 2 phosphatase. AB - Co-clustering of the type II receptors binding the Fc part of IgG (FcgammaRIIb) and B cell receptors results in the translocation of cytosolic, negative regulatory molecules to the phosphorylated immunoreceptor tyrosine-based inhibitory motif (P-ITIM) of the FcgammaRIIb. SH2 domain-containing protein tyrosine phosphatases (SHP-1 and SHP-2), and the polyphosphoinositol 5' phosphatase (SHIP) have been reported earlier to bind to murine FcgammaRIIb P ITIM. However, neither the functional substrates of these enzymes, nor the mechanism of the inhibition are fully resolved. We show here that the human FcgammaRIIb binds SHP-2 when co-clustered with the B cell receptors, whereas its synthetic P-ITIM peptide bindes SHP-2 and SHIP in lysates of the Burkitt's lymphoma cell line BL41. The P-ITIM peptide binding enhances SHP-2 activity, resulting in dephosphorylation and release of P-ITIM-bound SHIP and Shc. Moreover, P-ITIM-bound SHP-2 dephosphorylates synthetic peptides corresponding to the sites of tyrosine phosphorylation on SHIP and Shc, indicating that these proteins are its potential substrates. Thus SHP-2-induced dephosphorylation may modulate the intracellular localization and/or activity of SHIP and Shc, thereby inhibiting further activation pathways which they mediate. PMID- 10382762 TI - CD28-negative cytolytic effector T cells frequently express NK receptors and are present at variable proportions in circulating lymphocytes from healthy donors and melanoma patients. AB - In humans, NK receptors are expressed by natural killer cells and some T cells, the latter of which are preferentially alphabetaTCR+ CD8+ cytolytic T lymphocytes (CTL). In this study we analyzed the expression of nine NK receptors (p58.1, p58.2, p70, p140, ILT2, NKRP1A, ZIN176, CD94 and CD94/NKG2A) in PBL from both healthy donors and melanoma patients. The percentages of NK receptor-positive T cells (NKT cells) varied strongly, and this variation was more important between individual patients than between individual healthy donors. In all the individuals, the NKT cells were preferentially CD28-, and a significant correlation was found between the percentage of CD28- T cells and the percentage of NK receptor+ T cells. Based on these data and the known activated phenotype of CD28- T cells, we propose that the CD28- CD8+ T cell pool represents or contains the currently active CTL population, and that the frequent expression of NK receptors reflects regulatory mechanisms modulating the extent of CTL effector function. Preliminary results indicate that some tumor antigen-specific T cells may indeed be CD28- and express NK receptors in vivo. PMID- 10382763 TI - NILR-1, a novel immunoglobulin-like receptor expressed by neutrophilic granulocytes, is encoded by a leukocyte receptor gene complex on rat chromosome 1. AB - Several receptors expressed by subsets of leukocytes and with sequence homology to the killer cell inhibitory receptors have recently been identified both in man and mouse. Here we describe a rat cDNA that encodes a novel receptor of this group, designated neutrophil immunoglobulin-like receptor-1 (NILR-1). The predicted 58.7-kDa mature NILR-1 protein is a type I integral membrane protein, with three C2-type immunoglobulin superfamily domains, a transmembrane region devoid of charged amino acids, and a cytoplasmic tail containing four immunoreceptor tyrosine-based inhibition motif-like regions. NILR-1 shows greatest sequence homology to the mouse paired immunoglobulin-like receptor-B and members of the human leukocyte immunoglobulin-like receptor/immunoglobulin-like transcript group of receptors. As shown by Northern blot analysis, NILR-1 was transcribed by neutrophilic granulocytes. Although weaker transcription was found with a macrophage cell line, no signal was detected with peritoneal macrophage or spleen RNA. Linkage analysis localized Nilr1 to chromosome 1, closely linked to a locus encoding a rat NKp46 orthologue. The two loci define a rat leukocyte receptor gene complex, in a region syntenic to human chromosome 19q13.4 and the proximal part of mouse chromosome 7, that harbors the human and mouse leukocyte receptor gene complexes. PMID- 10382764 TI - Characterization of mouse interleukin-12 p40 homodimer binding to the interleukin 12 receptor subunits. AB - Interleukin-12 (IL-12) is a heterodimeric cytokine composed of two disulfide bonded subunits, p35 and p40, which has important regulatory effects on T cells and natural killer (NK) cells. In contrast to heterodimeric IL-12, a homodimer of the p40 subunit, designated (p40)2, has been shown to be a potent IL-12 antagonist. To study the interaction between (p40)2 and the known IL-12 receptor (IL-12R) subunits, IL-12Rbeta1 and IL-12Rbeta2, we directly measured the binding activity of mouse (p40)2 to ConA-activated lymphoblasts and purified B cells from splenocytes of C57BL/6J mice. These results demonstrated the presence of both high (Kd about 5 pM) and low affinity (Kd about 15 nM) binding sites for mouse 125I-labeled (p40)2. To elucidate which of the IL-12R subunits binds mouse (p40)2, binding studies of mouse 125I-labeled (p40)2 to Ba/F3 cells expressing recombinant mouse IL-12Rbeta1 and/or mouse IL-12Rbeta2 were carried out. Mouse IL 12Rbeta1 bound mouse 125I-labeled (p40)2 with high and low affinities, comparable to that observed on Con A blasts and B cells. In contrast, mouse IL-12Rbeta2 bound mouse 125I-labeled (p40)2 very poorly. These data demonstrate that similar to IL-12, mouse (p40)2 binds with both high and low affinity to Con A blasts and B cells, and that IL-12Rbeta1 is responsible for mediating the specific binding of mouse (p40)2. PMID- 10382765 TI - Immunization with alpha-galactosylceramide polarizes CD1-reactive NK T cells towards Th2 cytokine synthesis. AB - We have compared the immune responses of mice immunized either with alpha galactosylceramide (alpha-GalCer), capable of eliciting a CD1-metiated stimulation of V alpha14+ NK T cells, or with lipoarabinomannan (LAM), a glycophospholipid derived from mycobacteria which is known to be presented by CD1b in humans. Within 24 h, alpha-GalCer induces a burst of IFN-gamma secretion in vivo, and recall with antigen in vitro leads to the synthesis of IL-4 and IL 10 in addition to IFN-gamma. Associated with this in vivo cytokine release is a polyclonal activation of splenic B and T cells. CD1-reactive NK T lymphocytes mediate these events, because none of them are observed in alpha-GalCer-immunized CD1-/- mice. LAM immunization fails to promote similar early responses in vivo. Repeated exposure of mice to alpha-GalCer induces splenic T cells to secrete IL-4 and IL-10 but dramatically reduced levels of IFN-gamma. Such a bias in the cytokine balance triggered by NK T cells stimulated with multiple doses of alpha GalCer suggests that this compound might be useful in the induction of Th2 immune responses and the prevention of chronic inflammatory conditions mediated by Th1 cytokines. PMID- 10382766 TI - Induction of antibody responses to new B cell epitopes indicates vaccination character of allergen immunotherapy. AB - Whether the modulation of antibody responses can contribute to the improvement of clinical symptoms in patients receiving allergen immunotherapy represents a controversial issue. We have used purified [seven recombinant (r) and one natural] timothy grass pollen allergens as well as recombinant B cell epitope containing fragments of the major timothy grass pollen allergen, Phl p 1, to investigate humoral immune responses in eight allergic patients receiving grass pollen-specific immunotherapy. We found that the administration of aluminium hydroxide-adsorbed grass pollen extract induced complex changes in allergen/epitope-specific antibody responses: increases in IgG subclass (IgG1, IgG2, IgG4) responses against allergens recognized before the therapy were observed. All eight patients started to mount IgE and IgG4 responses to continuous Phl p 1 epitopes not recognized before the therapy and a de novo induction of IgE antibodies against new allergens was found in one patient. Evidence for a protective role of IgG antibodies specific for continuous Phl p 1 epitopes was provided by the demonstration that preincubation of rPhl p 1 with human serum containing therapy-induced Phl p 1-specific IgG inhibited rPhl p 1 induced histamine release from basophils of a grass pollen-allergic patient. Our finding that immunotherapy induced antibody responses against previously not recognized B cell epitopes indicates the vaccination character of this treatment. The fact that patients started to mount de novo IgE as well as protective IgG responses against epitopes may explain the unpredictability of specific immunotherapy performed with allergen extracts and emphasizes the need for novel forms of component-resolved immunotherapy. PMID- 10382767 TI - Switch in chemokine receptor expression upon TCR stimulation reveals novel homing potential for recently activated T cells. AB - When naive T lymphocytes are activated and differentiate into memory/effector cells, they down-regulate receptors for constitutive chemokines such as CXCR4 and CCR7 and acquire receptors for inflammatory chemokines such as CCR3, CCR5 and CXCR3, depending on the Th1/Th2 polarization. This switch in chemokine receptor usage leads to the acquisition of the capacity to migrate into inflamed tissues. Using RNase protection assays, staining with specific antibodies, and response to recombinant chemokines, we now show that following TCR stimulation, memory/effector T cells undergo a further and transient switch in receptor expression. CCR1, CCR2, CCR3, CCR5, CCR6 and CXCR3 are down-regulated within 6 h, while CCR7, CCR4, CCR8 and CXCR5 are up-regulated for 2 to 3 days. Up-regulation of CCR7 following TCR stimulation was observed also among resting peripheral blood T cells and required neither co-stimulation nor exogenous IL-2. On the other hand IL-2 down-regulated CXCR5, up-regulated CCR8 and facilitated the recovery of CCR3 and CCR5. Upon TCR stimulation, Th1 and Th2 cells produced comparable sets of chemokines, including RANTES, macrophage inflammatory protein 1beta, I-309, IL-8 and macrophage-derived chemokine, which may modulate surface chemokine receptors and contribute to cell recruitment at sites of antigenic recognition. Altogether these results show that following TCR stimulation effector/memory T cells transiently acquire responsiveness to constitutive chemokines. As a result, T cells that are activated in tissues may either recirculate to draining lymph nodes or migrate to nearby sites of organized ectopic lymphoid tissues. PMID- 10382768 TI - Genes in two major histocompatibility complex class I regions control selection, phenotype, and function of a rat Ly-49 natural killer cell subset. AB - We have generated a monoclonal antibody (STOK2) which reacts with an inhibitory MHC receptor on a subset of alloreactive NK cells in the rat. This receptor, termed the STOK2 antigen (Ag), belongs to the Ly-49 family of lectin-like molecules and displays specificity for the classical MHC class I molecule RT1-A1c of PVG rats. Here, we have investigated the influence of the MHC on the selection, phenotype and function of the STOK2+ NK subset in a panel of MHC congenic and intra-MHC recombinant strains. STOK2 receptor density was influenced by the presence of its classical MHC I ligand RT1-A1c, as evidenced by a reduction of STOK2 Ag on the surface of NK cells from RT1-A1c+, as compared with RT1-A1c-, strains. In addition, a role for nonclassical MHC I RT1-C/E/M alleles in the selection of the STOK2 Ag was demonstrated. The relative number of STOK2+ NK cells was fivefold higher in rats expressing the RT1-C/E/M(av1) as compared with those expressing the RT1-C/E/M(u) class Ib haplotype. The STOK2 ligand RT1 A1c inhibited cytotoxicity of STOK2+ NK cells regardless of effector cell MHC haplotype. Allospecificity of STOK2+ NK cells varied markedly with effector cell MHC, however, and suggested that inhibitory MHC I receptors apart from STOK2 were variably co-expressed by these cells. These data provide evidence for the MHC dependent regulation of the allospecific repertoire within a subset of potentially autoreactive Ly-49+ rat NK cells. PMID- 10382769 TI - Posterolateral intertransverse process spinal arthrodesis with rhBMP-2 in a nonhuman primate: important lessons learned regarding dose, carrier, and safety. AB - Recombinant osteoinductive proteins have been used successfully in canine and rabbit models of posterolateral intertransverse process arthrodesis, but little is known about the ability of these compounds to achieve fusion in nonhuman primates. The goals of this investigation were to compare different combinations of recombinant human bone morphogenetic protein-2 (rhBMP-2) dosages and carriers in a nonhuman primate model of posterolateral intertransverse process spinal fusion and to determine the feasibility of using rhBMP-2 in the presence of exposed dura in a laminectomy model. Posterolateral intertransverse process arthrodeses were performed at L4-5 in 29 rhesus monkeys. The most striking findings were as follows: rhBMP-2 could induce bone in a nonhuman primate spine; the presence of a laminectomy defect with exposed dura did not preclude the safe use of rhBMP-2 for posterolateral fusion; soft tissue compression of the collagen sponge carrier prevented bone induction at standard BMP doses, presumably due to squeezing of the protein out of the sponge; and longer rhBMP-2 loading time into the collagen carrier and mechanical protection from the soft tissue compression both allowed more bone induction at a lower dose of rhBMP-2. PMID- 10382770 TI - Risk factors associated with methicillin-resistant staphylococcal wound infection after spinal surgery. AB - We used the data from a retrospective case controlled study to identify risk factors for methicillin-resistant staphylococcal wound infection after spinal surgery. Thirty-five cases and 35 uninfected control patients were matched for indication for initial surgery and approximate operative date. Preoperative, intraoperative, and postoperative risk factors were examined. At our institution between 1989 and 1995, 35 adult patients developed spinal wound infection requiring operative debridement; 16 infections were caused by methicillin resistant staphylococci (MRS). Significant risk factors for MRS infection were lymphopenia, history of chronic infections, alcohol abuse, recent hospitalization, and prolonged postoperative wound drainage. Patients with MRS infections were also somewhat less likely to have received vancomycin prophylaxis. In contrast, the only factor associated with infection caused by other pathogens was alcohol abuse. A number of preoperative risk factors were significantly associated with subsequent MRS spinal wound infection. Chemoprophylaxis with vancomycin should be targeted to patients at increased risk, because overuse may promote the emergence of vancomycin-resistant pathogens. PMID- 10382771 TI - Outpatient laminotomy and discectomy. AB - This is a prospective study of 61 consecutive patients undergoing lumbar laminotomy and discectomy on an outpatient basis. The purpose of this study was to report on the feasibility of performing lumbar laminotomy and discectomy as an outpatient procedure and to assess perioperative complications, patient satisfaction, cost, and clinical results. Conventional lumbar laminotomy and discectomy traditionally requires a 1-3-day hospital stay. Recent advances in anesthesia and surgical techniques, as well as observation of patient progress after this procedure, has led the authors to believe that that this procedure may be performed on an outpatient basis without compromising patient satisfaction, outcome, or complications. Sixty-one consecutive patients underwent surgery for herniated nucleus pulposus in the lumbar spine. The procedure was performed under loupe magnification without the use of a microscope. Clinical outcome and patient satisfaction were assessed at an average follow-up of 12.5 months. The results showed 62% excellent, 31% good, 7% fair, and there were no reports of a poor outcome. During the time of the study, four patients (7%) were admitted to the hospital after the procedure for reasons of pain control, inability to void, or lack of caregiver at home. Overall cost savings were reflected in the cost of inpatient stay when compared to a representative group of inpatients. Laminotomy and discectomy for a hemiated nucleus pulposus has 93% good or excellent results as shown by this study and previous studies. Laminotomy and discectomy, which remains the gold-standard procedure for herniated disc surgery, can be performed safely and effectively as an outpatient procedure in the majority of patients. PMID- 10382772 TI - Factors predicting postoperative complications following spinal fusions in children with cerebral palsy. AB - A retrospective review of 107 patients with cerebral palsy who had undergone a posterior spinal fusion with unit rod instrumentation by the same two surgeons was done to determine what factors cause complications that lead to delayed recovery time and a longer than average hospital stay. The operative risk score was developed with scores for the child's ability to walk and talk, oral feeding ability, cognitive ability, and medical problems within the year prior to surgery. Operative risk score is primarily a measure of degree of neurologic involvement. The postoperative complication score (POCS) is a combined measure of all postoperative complications including factors for prolonged intubation, intensive care unit stay, hospital stay, and delayed feeding. The mean age at surgery was 14.3 years. The mean weight was 29.5 kg, with 89 of 107 patients below the fifth percentile for weight compared with age. The mean degree of spinal deformity was 75.2 degrees (range 43-120 degrees ). The mean weight for age was -1.96 SD below the normal. The mean operative time was 4.3 h, with estimated blood loss of 1.2 blood volumes. The mean length of hospitalization was 23 days 2 h, with 5 days 2 h in the intensive care unit. The operative risk score and weight for chronological age below the fifth percentile showed statistical significance (p = 0.05) in regard to increased POCS. The weight for height-age and deficient total lymphocyte count, both factors that measure nutritional status, showed no statistical significance (p > 0.05) compared with POCS. Curves with deformity of >70 degrees had statistically significant high POCS (p = 0.03). Complications for patients having a posterior and an anterior surgery versus those who had a posterior fusion alone were not statistically different (p > 0.05). The factors that led to a greater rate of complications were the severity of neurologic involvement, severity of recent history of significant medical problems, and severity of scoliosis. PMID- 10382773 TI - Simultaneous anterior and posterior approaches to the spine for revision surgery: current indications and techniques. AB - Revision spinal surgery often requires attention to both the anterior and the posterior portions of the spine. Staged, sequential, and more recently simultaneous anterior and posterior approaches have been proposed. A simultaneous approach has the distinct advantage of allowing complete and constant control of the anterior and posterior portions of the spine during surgery. The simultaneous approach has been shown to offer decreased operating time, blood loss, complication rate, and hospital length of stay as compared with staged procedures. The evolution in spinal instrumentation and ancillary equipment has greatly advanced the simultaneous technique. The development of a special operating table has facilitated patient positioning and intraoperative patient adjustments, optimizing operative exposure for the anterior and posterior surgical teams. The two-rod and four-rod techniques offer the surgeon the possibility to safely address complex deformities, particularly in kyphosis. PMID- 10382774 TI - Anterior cervical interbody fusion with plate fixation for chronic spondylotic radiculopathy: a 2- to 8-year follow-up. AB - In retrospectively analyzing 35 consecutive patients with chronic spondylotic radiculopathy treated by nerve root decompression, interbody fusion (Robinson technique), and plating, we studied the perioperative complication rate as well as the long-term clinical and radiologic outcomes of an additional plate fixation in degenerative cervical disorders. After an average of 54 months (range 24-102 months), all cases were reviewed for the purpose of this study. There were no perioperative or postoperative complications related to the plate fixation. In particular, there was no infection, graft extrusion, or neurologic deterioration. A solid fusion was obtained in all cases with a single-level fusion and in 87% of the cases with a multilevel fusion. The overall fusion rate was 94%. The clinical outcome of the patients with chronic radiculopathy was comparable with that in the literature, with only three patients (8.6%) having a poor result. This study demonstrated that plate fixation can be a useful adjunct in patients undergoing interbody fusion for cervical spondylotic radiculopathy. Plate fixation seems to reduce the rate of nonunion without additional hazards for the patient. This report should form the basis for a prospective randomized trial to answer the question more conclusively of whether an additional plate fixation is superior to uninstrumented cervical fusion in degenerative disorders. PMID- 10382775 TI - Thoracic pedicle screw placement guided by computed tomographic measurements. AB - Cadaveric pedicle screw placement guided by the measurements from axial computed tomography (CT) scans in the thoracic spine was assessed in this study. Axial CT scans were performed on four cadaveric thoracic spines, and the measurements included the pedicle transverse angle, inner pedicle width, and distance between the midline of the vertebra and the pedicle axis on the dorsal aspect of the lamina. With utilization of the data from CT scans, screws were directly placed into the thoracic pedicle from T1 to T10. Screw penetration of the pedicle was determined by gross examination. The results showed that the largest pedicle transverse angle was found at the levels of T1-2, and the smallest occurred at the T3 through T8 levels. The value of the pedicle inner width was quite different between specimens with a minimum of 3.0 mm at T4 and a maximum of 9.2 mm at T10. Gross examination of the pedicle showed that 13 (16.3%) of 80 screws penetrated the pedicle wall, with a Grade I penetration in 11 pedicles and a Grade II penetration in 2 pedicles. Screw penetration of the medial wall was found in four pedicles and penetration of the lateral wall was noted in nine pedicles. No screw penetration of the superior and inferior walls of the pedicle was identified in any of the four specimens. Thoracic pedicle screw placement guided by the measurements from axial CT scans significantly reduced the incidence of pedicle penetration. Axial CT measurements of the pedicle inner diameter and transverse angle as well as the starting point for screw insertion are recommended if pedicle screw fixation is intended in the thoracic spine. PMID- 10382776 TI - Pedicle screw placement at the sacrum: anatomical characterization and limitations at S1. AB - Anatomical and biomechanical data have suggested that pedicle screw fixation at the sacrum is optimum in the anteromedial direction into the S1 vertebral body, yet the possibility of posterior iliac crest interference with this screw pathway has been considered but not defined. This study aimed to determine if the anteromedial direction of screw placement into the vertebral body is possible in all cases at S1 and to assess the limiting effect of the posterior iliac crest. Computed tomography scans of the upper sacrum at the S1 pedicle parallel to the sacral endplate were examined in 100 patients. Analysis using a digitizer allowed characterization of an ideal screw pathway with variable screw and screw head diameters in an anteromedial direction into the S vertebral body. The effects of the posterior iliac crest upon these pathways were studied. The study demonstrated that anteromedial placement with bicortical fixation at the vertebral body was theoretically possible in almost all (98.5%) cases. Because the sacral body is often wider than the sacral spinal canal, a straight-ahead screw direction will often achieve placement into the S1 vertebral body, if the starting point for the screw allows screw placement adjacent to the medial border of the S1 pedicle with only 1.5 mm of cortical bone separating the canal and the screw. The space between the posterior iliac crest and the lateral aspect of the screw corridor ranges from a maximum of 52.4 mm to a minimum of 12.8, 6.2, and 0 mm for the 7-, 10-, and 12.5-mm screw corridors. On only three occasions (1.5%) was the ideal screw corridor not possible because of posterior iliac crest overlap. In each case, this occurred only unilaterally and when the widest of the screw corridors (12.5 mm) was used. Both the distance between the posterior iliac crests and the space available for optimum screw placement are greater in females than males. PMID- 10382777 TI - The USS pedicle hook system: a morphometric analysis of its safety in the thoracic spine. Universal Spine System. AB - The Universal Spine System (USS) pedicle hook design includes a fixation screw that passes obliquely in the anterocranial direction in the pedicle. The addition of the fixation screw was to address concerns with rotation of the hook and hook disengagement. This study was designed to evaluate the safety of the USS screw locked pedicle hook. Eleven cadaveric thoracic spines were instrumented posteriorly with USS pedicle hooks from T1 to T12. Spinal instrumentation was performed by a spinal surgeon experienced with the USS system. Spinal deformity was created prior to instrumentation, ranging from 0 to 55 degrees in the horizontal plane (rotation) and from 0 to 50 degrees in the frontal plane (scoliosis). Radiographs, computed tomography (CT), and segmental dissection were used for data acquisition. Morphometric CT analysis before instrumentation demonstrated that the transverse pedicular diameter was the smallest at T5 with a mean of 3.7 mm. The transverse pedicular angle (TPA) was found to always point toward the midline. The largest TPA was observed at T1 with a mean TPA of 28.4 degrees. The pedicle with the least angular deviation from the midline was T11 with a mean TPA of 7 degrees. Postinstrumentation CT analysis and segmental dissection revealed perforations of the pedicle cortex by the fixation screw in 15% of instrumented pedicles (26/172). There were 6 medial and 20 lateral perforations. Medial perforations occurred exclusively in the three most proximal spinal segments, whereas the lateral perforations occurred throughout the thoracic spine. The mean encroachment of the fixation screw was 1.67 mm medially and 1.95 mm laterally. This study demonstrates the variation in caliber and direction of the thoracic pedicles. Medial and lateral perforations of the pedicle can occur with the USS pedicle hook instrumented system. PMID- 10382778 TI - Segmental square spinal instrumentation for posterior lumbar spinal fixation. AB - The purpose of this study is to evaluate the clinical results of operations using a new spinal instrumentation for posterior fixation, called segmental square spinal (3-S) instrumentation. The 3-S instrument consists of two pairs of hooks to clasp the interarticular portion of the lamina bilaterally, two horizontal bars to connect the hooks on the right and left, and rods to connect the hooks side by side on the top and bottom. Twenty-seven patients who had degenerative lumbar disorders underwent operations with the 3-S instrument. Rigid fixation was obtained in all cases immediately after the operation. Bone union rates were 91% (21/23) with posterolateral fusion. The 3-S instrument seems to be suitable for spinal disorders in which anterior spinal fusion is not necessary, especially for degenerative spinal disorders. PMID- 10382779 TI - Up-regulated expression of matrilysin and neutrophil collagenase in human herniated discs. AB - Spontaneous resorption of herniated nucleus pulposus (HNP) is commonly observed when there is substantial contact of the disc with the spinal canal. We already demonstrated the expression of matrix metalloproteinase (MMP)-3 (stromelysin-1) in the granulation tissues of HNP, suggesting its role in the resorption process of HNP. Recent studies of osteoarthritic cartilages reported an up-regulated expression of metalloproteinases including MMP-7 (matrilysin) and MMP-8 (neutrophil collagenase), suggesting their roles in the matrix degradation. To clarify the expression of MMP-7 and MMP-8 in HNP, immunohistological analysis of various types of HNP was performed. We found MMP-7 was expressed in infiltrated mononuclear cells and chondrocytes, whereas MMP-8 was specifically expressed in chondrocytes. The positive rate for both MMP-7 and MMP-8 significantly increased when HNP was exposed to the epidural space (p < 0.01). Our data suggest that not only MMP-3 but also MMP-7 and MMP-8 may play a role in the resorption process of HNP. PMID- 10382780 TI - Ossification of the yellow ligament and spondylosis and/or ossification of the posterior longitudinal ligament of the thoracic and lumbar spine. AB - Combinations of varying degrees of spondylosis and/or ossification of the posterior longitudinal ligament (OPLL), and ossification of the yellow ligament (OYL) contribute to thoracic and lumbar neural compression in North Americans. Preoperative magnetic resonance and computed tomography examinations dictated the surgical approaches used to address spondylosis/OPLL in 11 patients, OYL in 12 patients, and spondylosis/OPLL and OYL in 3 patients. Myelopathy (4 patients), radiculopathy (13 patients), and cauda equina dysfunction (11 patients) were observed, 2 patients showing combined deficits. Outcomes (Odom's criteria) after laminectomy (24 patients) and circumferential thoracic procedures (2 patients) were good to excellent in the 73% of patients with spondylosis/OPLL, in 83% with OYL, and excellent for all 3 with spondylosis/OPLL and OYL. Full recognition of thoracic or lumbar spondylosis/OPLL and OYL ensure optimal surgical planning and outcomes. PMID- 10382781 TI - Balloon device for experimental graded spinal cord compression in the rat. AB - We have developed a balloon device that creates reproducible graded compression of the rat spinal cord. It uses a modified Camino intracranial pressure monitor bolt with a small latex balloon attached to the tip. The device is affixed to the spinous processes of two cervicothoracic vertebrae and positioned directly over an exposed segment of spinal cord. Ten compression balloons were tested and revealed reproducible pressure transmission at expansion volumes from 0.12 to 0.34 cc. Reversible graded spinal cord compression was verified by monitoring cortical somatosensory and motor evoked potentials before, during, and after cord compression. The pathophysiologic changes occurring with graded compression and the effect of therapeutic interventions can be studied in a rat model. PMID- 10382782 TI - A King type II curve pattern treated with selective thoracic fusion: case report with 44-year follow-up. AB - The optimal surgical treatment of the King-Moe type II thoracic curve pattern is controversial. The issue of postoperative "decompensation" has arisen in conjunction with the use of third-generation instrumentation systems. This report presents the 44-year clinical and radiographic follow-up of a patient with a type II scoliosis treated with uninstrumented selective thoracic fusion using the criteria of King and Moe. The caudal extent of the fusion was defined by proper identification of the neutral and stable vertebra. At final follow-up, the patient remained well balanced and essentially pain free. Her level of function was "above average" for her age, as per SF-36 evaluation. PMID- 10382783 TI - Associations of isometric and isoinertial trunk muscle strength measurements and lumbar paraspinal muscle cross-sectional areas. AB - The relationships of dynamic and static trunk muscle strength measurements and muscle geometry are studied. Physiologically, isometric muscle strength is directly related to muscle cross-sectional area. We measured isometric and isoinertial trunk muscle strength of 111 former elite male athletes, aged 45-68, by Isostation B-200. Paraspinal muscle cross-sectional areas were measured from axial magnetic resonance images at the L3-L4 level. Isometric and isoinertial torques were closely related, but angular velocities were not predicted by isometric maximal torque. The area of the psoas muscles correlated with isometric maximal flexion, as well as with isoinertial maximal torque. angular velocity, and power in flexion (r = 0.24-0.27). The area of the extensor group correlated with isometric maximal extension and with isoinertial maximal torque and power in extension (r = 0.24-0.25). We conclude that dynamic and static strength measurements are closely related, with angular velocity giving additional information on muscle function. Paraspinal muscle cross-sectional area is one determinant of isometric and isoinertial trunk muscle strength. PMID- 10382784 TI - Radiological study of cervical ossification of the posterior longitudinal ligament. AB - The rate of progression of cervical ossification of the posterior longitudinal ligament (OPLL) was radiologically studied during a 3-year period in three patient populations: (a) after laminoplasty (25 patients), (b) after laminectomy (16 patients), and (c) in patients who were managed without surgery (56 patients). There appeared to be no significant difference between these two surgical procedures in postoperative progression of OPLL. When progression of OPLL was compared between patients treated surgically and nonsurgically, posterior surgery accelerated progression of OPLL. PMID- 10382785 TI - Interleukin 1 receptor antagonist and E-selectin concentrations: a comparison in patients with severe acute pancreatitis and severe sepsis. AB - PURPOSE: This prospective clinical study was designed to compare interleukin 1 receptor antagonist (IL-1ra) and E-selectin concentrations in patients with severe acute pancreatitis to those with severe sepsis. MATERIALS AND METHODS: Nine consecutive patients with severe acute pancreatitis and 11 consecutive patients with severe sepsis admitted to a medical/surgical intensive care unit were included in the study. Plasma concentrations of IL-1ra and E-selectin were serially measured daily for 7 days or throughout their stay in the intensive care unit if shorter. RESULTS: The concentrations of IL-1ra were significantly higher on admission in patients with severe sepsis compared with the patients with severe pancreatitis (median levels 10,500 and 2,600 pg/mL, respectively, P = .007). When the data from the first 3 days were analyzed using analysis of variance (ANOVA), the levels of IL-1ra and E-selectin were similar in both groups. The concentrations of IL-1ra and E-selectin correlated to the development of multiorgan dysfunction as assessed by sequential organ failure assessment (SOFA) score (P = .032 and .043, respectively). CONCLUSION: This study shows that IL-1ra and E-selectin are released in acute severe pancreatitis, and the levels seem to be comparable to those in patients with severe sepsis. Concentrations of IL-1ra and E-selectin correlate to the development of multiorgan failure as indicated by high SOFA scores during the first week of disease. PMID- 10382786 TI - IL-1ra administration does not improve cardiac function in patients with severe sepsis. AB - PURPOSE: The purpose of this study was to investigate the effects of interleukin 1 receptor antagonist (IL-1ra) on myocardial function in septic patients. MATERIALS AND METHODS: A subgroup of patients from a prospective, randomized, double-blind, placebo-controlled, multicenter trial was studied from 63 academic medical centers in the United States, Canada, and Europe. A subgroup of 71 patients with severe sepsis in whom vasoactive support was little altered during the study was included. The patients were randomized to receive either placebo (n = 29) or IL-1ra at a dose of 1 mg/kg/h (n = 20) or 2 mg/kg/h (n = 22). RESULTS: Hemodynamic measurements were taken at baseline, and 1, 2, 3, 4, 8, and 12 hours after placebo or IL-1ra administration. No significant differences in hemodynamic parameters were observed between the groups or over time during the study period. CONCLUSIONS: IL-1ra administration has no effect on cardiac function in septic patients. PMID- 10382787 TI - Gut mucosal atrophy after a short enteral fasting period in critically ill patients. AB - PURPOSE: The purpose of this study was to evaluate the presence of gut mucosal atrophy and changes in mucosal permeability in critically ill patients after a short fasting period. MATERIALS AND METHODS: Fifteen critically ill patients underwent a period of enteral fasting of at least 4 days (mean 7.8 days). We took the following measurements the day before initiating enteral nutrition: indirect calorimetry, serum albumin, prealbumin, and lymphocyte count. We also performed a duodenal endoscopic biopsy with histopathological and mucosal morphometric analysis including villus height and crypt depth. The lactulose-mannitol test was performed to assess gut permeability. A total of 28 healthy volunteers served as controls for duodenal biopsy or lactulose-mannitol test. Clinical data, such as length of fasting, severity score, and previous parenteral nutritional support, were recorded. RESULTS: We found gut mucosal atrophy, expressed as a decrease in villus height and crypt depth, in patients compared with controls. The patients also exhibited an abnormal lactulose-mannitol test. Morphometric changes did not correlate with permeability. Further, we found no correlation between the results of the lactulose-mannitol test and of mucosal morphometry with clinical data. CONCLUSIONS: We found that a short period of enteral fasting was associated with significant duodenal mucosal atrophy and abnormal gut permeability in critically ill patients. PMID- 10382788 TI - Comparison between intrathoracic blood volume and cardiac filling pressures in the early phase of hemodynamic instability of patients with sepsis or septic shock. AB - PURPOSE: The purpose of this study was to analyze three different variables of cardiac preload; central venous pressure (CVP), pulmonary artery occlusion pressure (PAOP), and intrathoracic blood volume index (ITBVI) that served as the best indicator of cardiac function, that is, cardiac index (C1) or stroke index (SI). MATERIALS AND METHODS: This was a prospective study in 57 critically ill patients with sepsis or septic shock in whom 581 hemodynamic profiles were analyzed. One patient was included a second time after a period of 6 weeks. All patients were sedated and mechanically ventilated. Each patient had a 7.5-Frfive lumen pulmonary artery catheter (PAC) and a 4-Fr catheter with an integrated thermistor and fiberoptic that was advanced into the descending aorta via a femoral artery sheath. The study was performed in the surgical intensive care unit of a university hospital. RESULTS: Linear regression analysis of the first profile for each case (n = 58) revealed a significant correlation between ITBVI and SI (r = 0.66). For comparison, correlations for PAOP/SI (r = 0.06) and CVP/SI (r = 0.10) were poor. The analysis of all second profiles showed that only the change in ITBVI reflected the change in SI (r = 0.67), whereas PAOP (r = 0.07) and CVP (r = 0.05) failed. Furthermore, a positive change in SI (n = 265) was most often associated with an increase in ITBVI (n = 189, 71.3%), less for PAOP (n = 122, 46.0%) and CVP (n = 137, 51.7%). A reduction in SI (n = 256) was accompanied by a decrease in ITBVI (n = 176, 68.8%), PAOP (n = 119, 46.5%), and CVP (n = 118, 46.1%). An increase in ITBVI (n = 269) was accompanied by an increase in SI in 189 cases (70.3%). In these, PAOP increased only in 91 (48.1%) and CVP in 101 cases (53.4%), respectively. Accordingly, a positive change in PAOP (n = 218) was associated with an increase in SI in 122 cases (56.0%). ITBVI increased in 91 (74.6%) and CVP in 84 (68.9%) of these cases. A decrease in ITBVI (n = 250) was associated with a decrease in SI in 176 cases (70.4%). Decreases in PAOP (n = 89, 50.6%) and CVP (n = 91, 51.7%) did not reflect these changes. However, when PAOP (n = 229) and SI decreased (n = 119, 52.0%), ITBVI decreased in 89 (74.8%) and CVP in 73 cases (61.3%). CONCLUSIONS: In comparison with cardiac filling pressures, ITBVI seems to be the more reliable indicator of cardiac preload in patients with sepsis or septic shock. PMID- 10382789 TI - Partial liquid ventilation influences pulmonary histopathology in an animal model of acute lung injury. AB - PURPOSE: The aim of this study was to assess the effect of partial liquid ventilation (PLV) and conventional mechanical ventilation (CMV) in the pattern of distribution of lung injury in a rabbit model of acute lung injury. MATERIALS AND METHODS: Animals (1.5 to 3.5 kg) were assigned to receive CMV (tidal volume of 10 mL/kg and a PEEP of 5 cm H2O) or PLV with 18 mL/kg of intratracheal perflubron (tidal volume of 10 mL/kg and a PEEP of 5 cm H2O). Lung injury was elicited by intravenous administration of Escherichia coliendotoxin. Uninjured animals ventilated as the CMV group served as controls. After 4 hours of mechanical ventilation, the lungs were removed and tissue injury was assessed by light microscopy using a scoring system. RESULTS: Animals in the CMV group had higher lung injury scores in comparison to the PLV group (10+/-4.5 vs. 5+/-3.3, respectively, P < .05). The injury scores were similar for nondependent lung regions (CMV: 8+/-4.3, PLV: 6+/-2.9) but significantly different for the dependent regions (CMV: 12+/-4.6, PLV: 5+/-3.8, P< .05). CONCLUSIONS: PLV is associated with significant attenuation of lung injury, in comparison to CMV. This effect is predominantly due to attenuation of injury in the dependent region of the lung. PMID- 10382790 TI - A novel approach to monitor tissue perfusion: bladder mucosal PCO2, PO2, and pHi during ischemia and reperfusion. AB - PURPOSE: The purpose of this study is to determine if monitoring urinary bladder PCO2, PO2, and calculated intramucosal pH would be a reliable index of tissue perfusion. MATERIALS AND METHODS: This nonrandomized controlled study was conducted in a laboratory at a university medical center. Eight immature female Yorkshire pigs were studied with T-9 aortic cross-clamping for 30 minutes followed by a 60-minute period of reperfusion. Cystotomy was performed for placement of a Foley catheter and Paratrend 7 O2/CO2 sensor. RESULTS: Baseline hemodynamic and metabolic measurements were obtained along with measurements of bladder mucosal PO2 and PCO2 (mean+/-SEM). Blood flow measured with microspheres confirmed absence of blood flow during occlusion and hyperemia during reperfusion. Bladder mucosal PO2 decreased from 42+/-14.0 mm Hg (5.6 kPa) to 1.3+/-1.3 mm Hg (1.4 kPa) during the 30-minute interval of ischemia. This was followed by an increase of bladder PO2 to greater than baseline values at the end of the reperfusion period. Bladder mucosal Pco2 increased from 57+/-4.7 mm Hg (7.6 kPa) to 117+/-7.1 mm Hg (15.6 kPa) (P < .05) during ischemia. During reperfusion the Pco2 returned to baseline levels (55+/-4.0 mm Hg [7.3 kPa]). Calculated bladder mucosal pHi declined from 7.31+/-0.04 to 7.08+/-0.05 (P < .05) during the ischemic period and after reperfusion pHi was 7.17+/-0.03. CONCLUSIONS: Monitoring urinary bladder PO2, PCO2, or calculating pHi may provide a simple and reliable means of monitoring tissue perfusion. PMID- 10382791 TI - Influence of epinephrine and norepinephrine on intestinal villous blood flow during endotoxemia. AB - PURPOSE: The objective of this study was to determine the effects of epinephrine and norepinephrine on mucosal villous blood flow in a normotensive model of endotoxemia. MATERIALS AND METHODS: Thirty-two anesthetized rats were laparotomized, and a jejunal portion was exteriorized and opened by an antimesenteric incision. The jejunal segment was fixed on a plexiglass stage with the mucosal surface upward. Microcirculatory parameters were assessed by intravital videomicroscopy. The animals were randomly assigned to receive one of four treatments: infusion of Escherichia coli lipopolysaccharides (LPS, 2 mg/kg/h) without catecholamine pretreatment (LPS group); or infusion of LPS with epinephrine pretreatment (0.2 microg/kg/min, start 30 minutes before LPS infusion) (E group), or infusion of LPS with norepinephrine pretreatment (0.2 microg/kg/min, start 30 minutes before LPS infusion) (NE group). The control group did not receive either catecholamines or LPS. Mean diameter of central arterioles (D(A)) and mean erythrocyte velocity within the arterioles (V(E)) were measured 30 minutes before and at 0, 60, and 120 minutes after induction of endotoxemia. Mucosal villous blood flow was calculated from D(A) and V(E). RESULTS: LPS infusion alone and norepinephrine plus LPS infusion led to a significant vasoconstriction of central arterioles, which was associated with a similar decrease in mucosal villous blood flow. Epinephrine infusion alone led to a vasodilation and an increase in villous blood flow within the first 30 minutes. After induction of endotoxemia, D(A) returned to baseline values and villous blood flow was as low as in the LPS and the norepinephrine group after 120 minutes. CONCLUSION: In this experimental rat model, the catecholamines epinephrine and norepinephrine in a dosage of 0.2 microg.kg(-1).min(-1) neither diminish nor improve mucosal villous blood flow during the early phase of endotoxemia. PMID- 10382792 TI - Palatoalveolar outcome at 18 months following simultaneous primary cleft lip repair and posterior palatoplasty. AB - It is frequently reported that early repair of the soft palate induces narrowing of the remaining palatal cleft and thus facilitates later hard palate closure. However, to the best of our knowledge, there have been no comparative studies to test this hypothesis. The purpose of this retrospective study was to evaluate the change of palatoalveolar morphology following primary lip repair and posterior palatoplasty. Dental plaster models of patients with complete unilateral cleft of lip and palate (UCLP) were used to measure the width of the cleft and palatal arch. Twenty-six patients received simple posterior palatoplasty (PP group) simultaneous with primary lip repair, and 20 patients did not (NPP group). The dental models included one preoperative cast at 2 months (T1) and two or three casts at 6 (T2), 12 (T3), and 18 (T4) months before final palate closure. The linear measurements performed were width of alveolar cleft (Ca); width of palatal cleft between the canines (Cc), molars (Cm), and tuberosities (Ct); the palatal arch distance between the canines (Dc); the widest distance between molars (Dm) and the tuberosities (Dt); and the palatal height between the canines (Hc) and tuberosities (Ht). The raw measurements and the calculated cleft-to-arch ratios of Cc/Dc, Cm/Dm, and Ct/Dt were compared between the two groups. The results showed gradual narrowing of the width of cleft from T1 to T4. Narrowing of alveolar cleft width (Ca) from T1 to T2 was dramatic. The palatal arch (Dc, Dm, Dt) showed no change to mild increase in width. The cleft-to-arch ratios decreased with time. The palatal height remained the same or slightly increased over time. There were no significant differences observed between the PP and NPP groups among these measurements except for the Ct and Ct/Dt at T4. In conclusion, after initial lip repair, there was a decrease of the width of cleft in patients with complete UCLP during the 18-month period, and simple posterior palatoplasty did not further narrow the cleft nor influence palatal arch development. PMID- 10382793 TI - Vacuum-assisted closure in the treatment of degloving injuries. AB - Degloving injuries range from the occult, easily missed injury to obvious massive tissue damage. The serious nature of these wounds is exacerbated by mismanagement. It is generally accepted that the degloved tissue should be excised, defatted, fenestrated, and reapplied as a full-thickness skin graft. Dressings are required that provide gentle, evenly distributed pressure and avoid shear stress to the newly grafted skin. Numerous types of dressings have been devised but all are cumbersome and time-consuming. We have found the Vacuum Assisted Closure device to be a rapid, effective, and easy-to-use alternative to traditional methods. The authors examine their experience using a vacuum-assisted closure device to treat nine degloving injuries in 5 patients and discuss the important aspects in using this technique. PMID- 10382794 TI - Free flap reconstruction of the foot. AB - Free flap reconstruction of the foot has been widely performed in the last 20 years, but choice of a free transferred substitute for the soft tissue of the particular defect remains controversial. The authors present a series of 77 free flaps to the foot performed in 68 patients during October 1976 and September 1997. Long-term follow-up ranged from 12 months to 18 years (median, 44.4 months). Seventy-three flaps were transferred successfully (95%). The indications for a specific flap depended on the localization and extension of the foot defect. In weight-bearing areas the authors favored the use of a muscle flap covered with a split-thickness skin graft; the latissimus dorsi muscle was used primarily. This study shows a lower ulceration rate in muscle flaps covered with split-thickness skin grafts than in fasciocutaneous flaps in weight-bearing areas (27% vs. 60%). In nonweight-bearing areas, fasciocutaneous flaps were the best choice. In this series, the lateral arm flap was applied most often. The authors recommend free fascial flaps (serratus fascial flap or radial forearm fascial flap) covered by split- or full-thickness skin grafts for coverage of the malleolar region as well as coverage of exposed tendons of nonweight-bearing regions. Proper tailoring of the flap and postoperative care are very important to maintain a result without ulceration, as is avoiding having the suture line cross a weight-bearing area. Tactile sensation does not seem to be essential. PMID- 10382795 TI - Consideration of the UltraPulse carbon dioxide laser as a tool for skin deepithelialization. AB - Laserbrasion is essentially skin deepithelialization. The continuous-wave carbon dioxide (CO2) laser has a long history during which this capability to ablate the epidermis has been used in multiple clinical applications. The "improved" UltraPulse CO2 laser has been advocated as a safer method, primarily for skin resurfacing. The authors show in Sprague-Dawley rats by gross and histological examination that the UltraPulse CO2 laser can also be used effectively to achieve skin deepithelialization, with efficacy in clinical simulations without untoward effects on wound healing. The advantage of the UltraPulse CO2 laser appears to be less destruction to surrounding tissues. PMID- 10382797 TI - The Michigan Hand Outcomes Questionnaire (MHQ): assessment of responsiveness to clinical change. AB - Responsiveness is an important property of an outcomes questionnaire. It can be defined as the ability of an instrument to capture important changes in a patient's health status over time. The authors previously designed the Michigan Hand Outcomes Questionnaire (MHQ), a hand-specific outcomes instrument that contains six distinct scales: (1) overall hand function, (2) activities of daily living, (3) pain, (4) work performance, (5) aesthetics, and (6) patient satisfaction with hand function. In the first study, the authors demonstrated that the MHQ is a reliable and valid instrument for the hand. The purpose of this second study is to assess the responsiveness, or sensitivity, of the MHQ to clinical change in patient status. A total of 187 consecutive patients with chronic hand disorders completed a baseline MHQ prior to receiving treatment at a university plastic surgery clinic. Approximately 6 to 18 months after completing the first questionnaire, patients were sent a follow-up MHQ by mail. The second questionnaire was identical to the first, with the exception of one additional question added to each of the six MHQ scales. This additional question asked patients to rate the change in their hands since completing the last questionnaire using a seven-point response scale. Spearman's correlation coefficient was used to correlate the responses from patients' self-assessment questions with the actual score change (after score - before score). The response rate for the second administration was 49% (92 questionnaires returned)-a fairly good rate of return for mail surveys. There were no significant differences in gender, race, education, and income between responders and nonresponders. When patients' self-assessment of change was correlated with the change in the six scale scores over time, all six correlations were statistically significant, with p < 0.05. The correlations ranged from 0.25 for the aesthetics scale to 0.43 for the pain scale. The MHQ was responsive using patients' self-assessment of their clinical change. Future studies will evaluate the responsiveness of the MHQ compared with objective physiological measures such as grip strength, range of motion, and the Jebson-Taylor test. Additionally, research is underway to assess the responsiveness of the MHQ for specific procedures, including metacarpophalangeal arthroplasties for rheumatoid arthritis and microvascular toe to-hand reconstructions. PMID- 10382796 TI - Utility of vibration thresholds in patients with brachial plexus nerve compression. AB - The diagnosis of brachial plexus nerve compression is controversial due to the subjective nature of patient symptoms and the lack of objective, quantifiable tests. It has been hypothesized that quantitative sensory evaluation of sensory threshold is the most sensitive method of evaluating nerve compression, particularly in the early stages. This study evaluated the sensitivity and specificity of vibration thresholds for detection of brachial plexus nerve compression. A multiple-frequency vibrometer was used to evaluate 40 control subjects and 35 patients with brachial plexus nerve compression. Calculated sensitivity values were modest (0.49 at 63, 250, and 500 cps) with high specificity values (0.98 at 8 cps) for individual frequencies using a fifth percentile criterion. The low sensitivity values indicate that this instrument is not adequate as a screening device. PMID- 10382798 TI - Septic arthritis of the metacarpophalangeal and interphalangeal joints of the hand. AB - Septic arthritis of the small joints of the hand is not reported frequently. Twenty-eight patients were treated during a 15-month period, from November 1992 to January 1994. Prompt surgical drainage, antibiotics, and early postoperative mobilization (within 24 hours) were the cornerstones of management. Nineteen patients were available for a median follow-up of 10 months. Patients who presented after 10 days all had a poor result. Patients who presented earlier achieved a good result unless they defaulted from treatment or had sustained an associated severe traumatic joint injury. Ten of the 19 patients obtained a good result. Good functional results can be obtained with early postoperative mobilization. PMID- 10382799 TI - Gracilis muscle: arterial and neural basis for subdivision. AB - The gracilis muscle is commonly utilized by reconstructive surgeons in a variety of applications as a pedicled muscle or musculocutaneous flap, and as a free tissue transfer for soft-tissue coverage or as a functioning muscle transfer. The muscle anatomy has been well documented in the past. The aim of the present study was to study comprehensively the intramuscular neurovascular anatomy as it relates to segmental neurovascular functioning muscle transfer. The study was carried out in a series of 14 human cadavers. Each cadaver was injected with a lead oxide, gelatin, and water solution through the femoral arteries (200 ml per kilogram). The overall length of the musculotendinous unit was 44 +/- 2 cm, and the tendon comprised up to 6 +/- 2 cm of the length. The main arterial supply to the muscle entered 10 +/- 1 cm from the attachment to the body and inferior ramus of the pubis (diameter, 1.5-2.5 mm). The distal portion of the muscle was supplied by one to three small arterial branches of the superficial femoral artery. Venous drainage was noted to be through paired venae comitantes. The motor nerve arises from the obturator nerve and enters the muscle in association with the major vascular pedicle. The nerve then splits within the muscle and runs longitudinally in two or three major branches within the muscle parallel to the arterial branches and muscle fibers. The neurovascular anatomy of the gracilis muscle was found to be remarkably consistent from specimen to specimen, varying only in the length of the muscle and tendon, and the number of minor pedicles supplying the distal portion of the muscle. This study confirms the suitability of the gracilis for segmental functional muscle transfer. PMID- 10382800 TI - Repair of mild umbilical hernia. AB - A mild, persistent umbilical hernia that does not cause any functional problem is often ignored. The authors have devised a new technique to treat the mild, protrusive deformity of the umbilicus without associated complications. In this report, the new operative procedure is introduced. The authors have treated 72 patients with this method and have obtained good results. PMID- 10382801 TI - Modifications in endoscopic facelifts. AB - Since 1996, 72 patients (66 women and 6 men) have undergone endoscopic upper and midface rejuvenation. Sixteen of these patients had concomitant lower face rejuvenation at the same time. The patients were operated using a personal endoscopic technique, including biplanar endoscopic dissection (subgaleal and subcutaneous) at the forehead and temporal regions, excision of a galeal strip approximately 1 cm in thickness (to achieve a stable forehead lift), and a lower blepharoplasty incision for midface lifting and fixation of the malar fat pad. This approach helps to prevent midface widening-a concern of most surgeons. Regarding patient satisfaction, 51 patients had excellent results, 16 patients had good results, and 5 patients had an improved appearance of the mid and upper face. Complications included 11 incidences of temporary numbness in different regions of the upper and mid face, three incidences of temporary lower lid retraction (which did not require additional revision), three relapses of eyebrow elevation (which were reoperated), and unbalanced eyebrows in 2 patients (which were corrected during secondary revisional procedures). PMID- 10382802 TI - Reverse U-shaped split temporalis musculofascial flap in cranial base reconstruction. AB - To extend the versatility and range of the temporalis muscle, a new type of temporalis musculofascial flap was developed. This was achieved by dividing the muscle into two portions--anterior and posterior-while maintaining vascular communication between the deep and the middle temporal arteries. This flap is reverse U-shaped with one of the arms of the "U" corresponding to a pedicle, which supplies the blood, and the other corresponding to the recipient region. The bottom of the U corresponds to continuity between the anterior and posterior portions of the muscle, which contains the vascular communication. In two patients with meningioma, the flap was applied to occupy the extradural dead space combined with a pericranial flap to prevent leakage of cerebral spinal fluid to the dural defect. The reverse U-shaped split temporalis musculofascial flap has some advantages for intracranial reconstruction: sufficient rotational arc, adequate thickness, and rich vascularity. A reverse U-shaped split temporalis musculofascial flap is useful and of benefit, especially for reconstruction at the region of the anterior midline skull base. PMID- 10382803 TI - A new predictive modality of cranial bone thickness. AB - The objective of this study was to evaluate A-mode ultrasound in the assessment of cranial bone thickness utilizing an in vivo animal model. A prospective study was performed that identified four standardized calvarial points in 10 Landrace porcine skulls. The individual points were scanned with an A-mode ultrasonic transducer to obtain bone thickness measurements. The same points were measured subsequently using digital calipers for objective comparison. The accuracy of each of the measurement modalities was evaluated for inter- and intrarater reliability. The association between ultrasonic and caliper measurements was evaluated using Student's t-test, Pearson's correlation coefficient, and linear regression models to assess the effect of confounding variables. The mean difference between the ultrasonic and the caliper values was 0.31 +/- 0.22 mm (standard deviation). The statistical analyses employed strongly supported the predictive value of ultrasound as a function of the true calvarial thickness (p < 0.05, r > 0.88, R2 = 0.89). The results suggest that ultrasound is an accurate reflection of cranial bone thickness in an in vivo animal skull model. The development of a portable, noninvasive ultrasonic device can have substantial clinical implications for craniomaxillofacial surgery. PMID- 10382805 TI - Influence of radiation on late complications in patients with free fibular flaps for mandibular reconstruction. AB - Carcinoma of the mandible is a disease that evokes images of devastating functional and cosmetic outcomes. Most of these malignancies require treatment with surgical resection and perioperative irradiation (XRT). To minimize the incidence of postoperative complications, the timing of perioperative XRT has been questioned. This study reviewed 140 patients at M.D. Anderson Cancer Center over a 7-year period who underwent mandibular resection and reconstruction with a free fibular flap. The patients were divided into the following four groups: (1) preoperative XRT followed by immediate reconstruction, (2) preoperative XRT followed by delayed reconstruction, (3) postoperative XRT, and (4) no XRT. The complications studied included exposure of bone and hardware, orocutaneous fistula, osteoradionecrosis, partial and complete flap loss, and severe cervical contractures. Of the 140 patients studied, 59 (42%) had at least one complication. Complications per group were the following: group 1, 45%; group 2, 46%; group 3, 47%; and group 4, 28%. The results show that the incidence of complications is relatively equal between groups that received preoperative vs. postoperative XRT. PMID- 10382804 TI - Esophageal reconstruction by jejunal transfer. AB - Forty-five esophageal reconstructions were carried out using a free or pedicled transfer of jejunal segments. Thirty were performed for benign stenosis or atresia, and 13 were performed for malignancy. The remaining two were used to repair anastomotic leakage or fistulization resulting from prior esophagogastrostomy. Average anastomotic arterial and venous diameters were 1.2 mm and 3.0 mm respectively. Of the 45 reconstructions, 44 were successful; the single failure was the result of tearing of the mesenteric arcade. There were four fatalities. Jejunal transfer is an effective method of esophageal reconstruction. PMID- 10382806 TI - Evaluation of results and treatment variables for pressure ulcers in 48 veteran spinal cord-injured patients. AB - This retrospective study of 48 spinal cord-injured patients with pressure ulcers seen at a tertiary referral Veterans Hospital spinal cord injury unit between 1992 and 1997 correlates a number of variables (co-morbid conditions, nutritional status, smoking history, type of repair performed, type of bed used postoperatively, ulcer location and severity, duration of postoperative antibiotic therapy, time elapsed before sitting rehabilitation began, and length of hospital stay) with ulcer repair outcome measures, including postoperative systemic and wound-healing complications, recurrence rates, and the development of new ulcers at different sites. Surgical complication rates were high, occurring in 19 patients (39.6%), and ulcer recurrence or new ulcer development occurred in 38 patients (79.2%). Correlations were found between ulcer location and postoperative wound separation and the length of hospitalization. The hospital course was shorter if the ulcer was new rather than recurrent. Other than the finding that chronic smokers had longer courses of antibiotic therapy, smoking did not correlate statistically with other outcome variables, including wound-healing complications. No significant correlations were found between any postoperative systemic or wound complications, ulcer recurrence, or new ulcer development and patient age, level of spinal cord injury, number of ulcers and grade, laboratory values, mental status, cardiac or pulmonary disease, diabetes, and presence or absence of osteomyelitis. PMID- 10382807 TI - Stabilization of fibrin-chondrocyte constructs for cartilage reconstruction. AB - Cartilage replacement is a challenging issue in reconstructive surgery. In the past few years, tissue engineering has been tested as a means of cartilage reconstruction. Tissue engineering of cartilage depends on the use of adequate polymers. In addition to several natural and synthetic polymers, fibrin gel has been tested for cartilage reconstruction. However, fibrin is intrinsically unstable. The purpose of this study was to stabilize fibrin by increased fibrinolytic inhibition and to test these preparations for cartilage reconstruction with human nasal septum chondrocytes. Increased fibrinolytic inhibition was achieved with aprotinin and tranexamic acid. Stabilized fibrin chondrocyte constructs were cultivated for 4 weeks in vitro and compared with constructs made of standard, commercially available fibrin gel. The effect of several cell densities on stability, and the production of extracellular matrix components, were assessed on the basis of histology and immunohistochemistry. In contrast to constructs made of standard fibrin gel, stabilized constructs were stable for the entire observation period and demonstrated no or only minor shrinkage. Cells in these constructs appeared to be viable, and an extracellular matrix could be demonstrated in all constructs. The authors conclude that fibrin chondrocyte constructs stabilized by increased fibrinolytic inhibition could be an adequate tool for cartilage reconstruction. PMID- 10382808 TI - Lymphoscintigraphy with sentinel lymph node biopsy in cutaneous Merkel cell carcinoma. AB - Merkel cell carcinoma (MCC) is a rare cutaneous malignancy characterized by an aggressive clinical behavior with high rates of locoregional and systemic recurrence. Regional disease and distant metastases are associated with poor prognosis. Despite a predisposition of MCC to spread via the lymphatics, prophylactic lymph node dissection in the absence of clinically apparent lymph node involvement is controversial. The value of lymphoscintigraphy in cutaneous melanoma is established in lesions with ambiguous lymphatic drainage patterns. When used with sentinel lymph node biopsy (SLNB), it can identify subjects with occult regional node metastasis. The authors present 2 patients with MCC who underwent regional node staging with lymphoscintigraphy-directed SLNB. Both patients had sentinel nodes that were positive for metastatic disease. In patients with MCC, minimally invasive regional node staging SLNB may be useful in limiting the sequelae of routine lymphadenectomies. Whether early identification and treatment of patients with occult regional node disease can influence survival in MCC is not known. PMID- 10382809 TI - Extent of surgical intervention in primary soft-tissue aspergillosis. AB - Primary invasive Aspergillus Infection of the soft tissue is rare and typically affects immunocompromised patients in several distinct patterns of clinical presentation. In general, the role of surgery in the treatment of this disease is the removal of infected or necrotic tissue to prevent dissemination and mortality. However, the specific surgical recommendations have varied widely among reports due to the varied clinical circumstances in each series. The authors present the case of a patient with a primary invasive Aspergillus infection. They review the reported surgical experience with this disease, and discuss outcomes and surgical approaches in the context of several variations in clinical presentation. In all situations, antifungal therapy and prompt surgical intervention are critical in treating these initially localized but potentially lethal infections. The extent of intervention can range from minor debridement to amputation, and is based on the presence of persistent immunocompromise, the presence and extent of tissue necrosis, and the rate of progression during antifungal therapy. PMID- 10382810 TI - Reconstruction of full-thickness lower eyelid defects using a blepharoplasty technique with a hard palate mucosal graft. AB - The authors report a method of reconstruction of a full-thickness lower eyelid defect using a blepharoplasty technique utilizing excess skin of the lower eyelid with a hard palate mucosal graft. In all patients the hard palate mucosa took well, and good functional and aesthetic results were obtained. The most suitable indication of this technique might be for the defect occurring horizontally (for which direct closure is difficult to apply) and located in the lateral side of the lower eyelid (when a large amount of excess skin can be utilized). The authors conclude that although the shape and size of the defect to which this method can be applied is restricted, this is a useful option in the reconstructive methods of a full-thickness lower eyelid defect. PMID- 10382811 TI - Surgical correction of cryptotia using Medpor. AB - Cryptotia is a congenital auricular anomaly found more commonly in Orientals than whites. The characteristics of cryptotia are the invagination of the upper part of the auricle under the temporal skin and the deformity of the auricular cartilage. The goals of the repair of cryptotia are to release the upper ear from the side of the head to restore the retroauricular groove, to correct the malposition, and to correct the cartilaginous deformity. To lengthen the skin between the superior portion of the auricle and the scalp, the authors used both the modified Z-plasty and the temporal advancement flap. We partially detached the abnormal insertion of the superior auricular muscle at the upper part of the helix to make it weak. After complete exposure of the posterior aspect of the upper auricular cartilage, the constricted intrinsic transverse and oblique muscles were cut, and everting horizontal mattress sutures were inserted on the antihelix to expand the constricted body and crus of the antihelix. Thereafter, an ultrathin Medpor sheet (0.85-mm thickness) was fixed with 6-0 nylon sutures to the posterior aspect of the corrected antihelical cartilage for lengthening and splinting the relatively shortened upper pole of the deformed cartilage. This operative method is thought to be useful in maintaining the lengthened auricular height and shape, and in preventing the relapse of ear cartilage deformities. PMID- 10382812 TI - Fasciosubcutaneous flaps in reconstruction of lower extremity distal defects. PMID- 10382813 TI - Unusually shaped epidermoid cyst. PMID- 10382814 TI - Application of a sartorius muscle flap during abdominal wall reconstruction. PMID- 10382815 TI - One-stage coverage of a large anterior chest wall defect with multiple flaps in an octogenerian with recurrent sternal chondrosarcoma. PMID- 10382816 TI - Evaluation of a fully automated glycoprotein G-2 based assay for the detection of HSV-2 specific IgG antibodies in serum and plasma. AB - BACKGROUND: Ranking after infections with Chlamydia trachomatis and human papillomavirus, genital herpesvirus is the third most common sexually transmitted disease. The majority of recurrent genital herpes infections are caused by herpes simplex virus type-2 (HSV-2). Seroepidemiological studies on the prevalence of HSV-2 specific IgG antibodies are especially important to study the impact of this infection among risk groups. OBJECTIVE: To evaluate the sensitivity and specificity of the Cobas Core HSV-2 IgG specific assay (available for research use only), that can be run on the Cobas Core fully automated immuno-analyzer. STUDY DESIGN: The Cobas Core HSV-2 specific IgG EIA is based on macro-beads coated with affinity purified glycoprotein G-2 antigen from HSV-2 infected cells. The Cobas Core HSV-2 IgG specific assay was compared with the Chiron rapid immunoblot assay (RIBA), the Gull enzyme-linked immunosorbent assay (EIA) and the Centocor EIA. The respective assays were tested, using 1219 serum samples, from 612 females and 607 males attending the outpatient clinic for sexually transmitted diseases of the Erasmus Medical Center Rotterdam (EMCR). RESULTS: The consensus value, obtained by a concordant result with three out of four assays, demonstrated 350 positive samples (28.7%), 851 negative samples (69.8%) and 18 (1.5%) serum samples with a discordant result. The overall agreement of the Cobas Core HSV-2 EIA against the consensus value was 95.8% and the sensitivity and specificity proved to be 100 and 97.1% respectively. CONCLUSION: The results obtained with the Cobas Core HSV-2 EIA indicate that this is a useful, specific and sensitive assay for the detection of HSV-2 specific IgG antibodies in serum. The advantage of the Cobas Core HVS-2 EIA compared to the other assays, is that this assay can be performed in a fully automated process. PMID- 10382817 TI - Measles virus infection causes transient depletion of activated T cells from peripheral circulation. AB - BACKGROUND: Natural measles virus infection as well as vaccination with attenuated measles virus induce temporary immunosuppression, which is responsible for part of the morbidity and mortality associated with measles. The underlying molecular mechanisms are not known. Recently, in vitro studies have revealed a marked increase of LFA-1 expression of lymphocytes in the presence of infectious measles virus. OBJECTIVES: In order to further investigate immune dysfunction in measles we analyzed the expression of leukocyte function-associated antigen 1 (LFA-1) on ex vivo derived circulating human T cells. STUDY DESIGN: Expression of LFA-1 was measured by flow cytometry using monoclonal antibodies directed against CD11a and CD18. LFA-1 expression was followed in the course of infection in four adult seronegative vaccinees and in four patients with natural measles. RESULTS: There was a remarkable loss of LFA-1-bright cells during natural measles and after measles vaccination. The number of LFA-1-bright cells reached a minimum on day 7-14 after vaccination, and at the onset of rash during natural measles infection, and approached normal levels within 3-5 weeks. CONCLUSION: It is suggested that measles virus infection interferes with lymphocyte trafficking and reallocation. Disruption of recirculation and random homing of lymphocytes might contribute to the immunosuppression, which is characteristic for measles virus infection. PMID- 10382818 TI - A population-based prospective survey of newborn infants with suspected systemic infection: occurrence of sporadic enterovirus and adenovirus infections. AB - BACKGROUND: Enterovirus outbreaks are known to occur in neonatal wards and enteroviruses may cause community-acquired sepsis-like disease in the neonatal period. Less well is known their possible role in suspected systemic infections during the perinatal period. OBJECTIVES: To investigate the occurrence of enterovirus infections in neonatal patients suspected of systemic infection. STUDY DESIGN: A population-based prospective survey was organized in the hospitals of the Greater Helsinki Region during 13 months in 1993-94. Criteria for enrollment included onset of symptoms before the age of 29 days and a decision, on clinical grounds, to take a blood culture for bacteria. Acute phase samples of blood, feces, nasopharyngeal swab, and cerebrospinal fluid, if available, were inoculated in monolayer cultures of four different cell lines. In addition, enterovirus infections were searched for using an enterovirus group reacting IgM test. RESULTS: One hundred and thirty-seven patients had a sufficient number of specimens examined, and were thus evaluable. Most of the infants had the onset of the symptoms within a few days after birth. An enterovirus was isolated from four newborn infants (3%), while seven children (5%) were found to excrete adenovirus. Enteroviral antigen was detected in cell cultures inoculated with specimens from two additional infants. Virus-positive infants had no evidence of bacterial infection and did not show specific clinical signs or symptoms differentiating them from the rest of the study group. All enrolled infants recovered without sequelae. CONCLUSION: We conclude that sporadic viral infections may be common in neonatal patients with suspected systemic infection, and this should be taken into account when judging the etiology. PMID- 10382819 TI - Evaluation of BC-3 and BCP-1 cell lines in immunofluorescence assay to detect HHV 8 antibodies in Kaposi's sarcoma, multiple myeloma and other groups. AB - BACKGROUND: We describe a comparative study of an immunofluorescence assay using inducible BC-3 and BCP-1 cell lines as sources of HHV-8 antigens. STUDY DESIGN: Detection of both antibodies to proteins expressed in lytic cycle and during latency in sera from HIV-infected patients with KS, HIV-positive patients without KS, normal blood donors, HIV-negative pregnant women and HIV-negative patients with multiple myeloma. Where possible, detection of antibody was associated with nested PCR detection of HHV-8 in peripheral mononuclear cell (PBMC) samples collected from AIDS-KS patients. RESULTS: Immunofluorescence was more intense with the BC-3 cell line than with BCP-1, thus facilitating examination under the microscope. HHV-8 antibodies were detected among 82.75% of AIDS-KS patients, in 27.3% of HIV-infected homosexual men, 2% of blood donors and in 2% of pregnant women. No HHV-8 antibodies were detected in serum samples from HIV-negative patients presenting multiple myeloma. HHV-8 DNA sequences were detected and confirmed by southern blot hybridization in five out of 17 (29.4%) PBMC samples from AIDS-KS patients. Titre of antibodies to proteins expressed in lytic cycle was much higher than the titre of antibodies to proteins expressed during latency. CONCLUSIONS: Both immunofluorescence assays were found useful and HHV-8 seroprevalence rates reported in previous studies were confirmed. In addition, results obtained using these assays tend to provide evidence for a lack of epidemiological association between HHV-8 infection and development of multiple myeloma. PMID- 10382820 TI - Direct in situ reverse transcriptase-linked polymerase chain reaction with biotinylated primers for the detection of hepatitis C virus RNA in liver biopsies. AB - To assess the presence and the cellular distribution of hepatitis C virus (HCV) RNA in the liver of 11 patients with confirmed HCV infection, a direct in situ reverse transcriptase-linked polymerase chain reaction (RT-PCR) method was performed on formalin-fixed and paraffin-embedded biopsies. The oligonucleotide primers used were specific to the 5' non coding region. An unlabelled downstream oligonucleotide served as a primer for reverse transcription as well as PCR. The upstream oligonucleotide serving as a primer for PCR was biotinylated, allowing a direct enzymatic detection of PCR products. HCV infected cells revealed cytoplasmic staining mainly concentrated towards the interface of the nucleus and cytoplasm. Most of the stained cells were hepatocytes and sometimes Kupffer cells. The results were compared with those obtained by RT-PCR of RNA extracted from the corresponding tissue block. Extracted HCV RNA could be detected in liver tissues of nine out of 11 (82%) infected patients. The detection rate using in situ RT-PCR was 7/11 (63%). The use of labelled primers improved specificity of direct in situ methods, by preventing non-specific incorporation of labelled dNTPs into fragmented DNA. Further studies are however required in order to increase detection sensitivity of HCV infection by in situ molecular methods. PMID- 10382822 TI - Synthesis of functionally graded CO3 apatite as surface biodegradable crystals. AB - Apatite, which will have a degradable surface and a relatively stable core, was synthesized at 80+/-1 degrees C and pH 7.4+/-0.2 using a specially generated gradient carbonate supply system. X-ray diffraction analysis showed that the (300) reflection peak shifted to the higher angle direction due to the substitution of CO3(2-) ions into the PO4(3-) positions and was broader than that of homogeneous hydroxyapatite. Scanning electron microscopy indicated that the apatite was composed of small coagulated ellipsoidal crystals differing from needle-like crystals with hexagonal section of hydroxyapatite. Electron spectroscopy for chemical analysis showed a negative gradient of carbonate concentration with depth in the crystals. The apparent solubility in 0.5 moll(-1) acetate buffer solution (37 degrees C, pH 4.0) was much higher than that of homogeneous hydroxyapatite. The residual sample showed slim needle-like crystals. These results suggested that graded carbonate-containing apatite, namely CO3 apatite with solubility gradient in its crystal structure, was formed by this process and may be useful from the view point of surface biodegradable materials as bone representatives. PMID- 10382821 TI - The use of peptides from glycoproteins G-2 and D-1 for detecting herpes simplex virus type 2 and type-common antibodies. AB - BACKGROUND: identification and discrimination of latent herpes simplex virus (HSV) infection relies on antibody identification. The inclusion of synthetic peptides with HSV glycoproteins provides means for stable and discriminatory assays for population studies. OBJECTIVE: to determine whether virus-specific synthetic peptides might identify HSV type 2 (HSV-2) antibodies in the presence of the cross-reactive and more common HSV type 1 (HSV-1) antibodies. STUDY DESIGN: the capacity of synthetic peptides as HSV antigens was analyzed in enzyme immunoassay (EIA) using well characterized human serum cohorts. The HSV peptide assays were evaluated in comparison with two commercial HSV-2 assays. RESULTS: a combination of two C-terminal HSV-1 glycoprotein D (gD-1) peptides detected type common HSV immunoglobulin G (IgG) with high sensitivity (95%) and specificity (93%). Peptides derived from the C-terminus of HSV-2 glycoprotein G (gG-2) had a high HSV-2 type-specificity. Inclusion of both gD-1 and gG-2 peptides gave a sensitivity for human anti-HSV-2 IgG that was similar to that of assays including different amounts of native gG-2. With western blotting as a standard, the sensitivity of the peptide assay ranged between 86% for HSV-2 seropositive persons and 61% for HSV-2 seroconverters. Addition of a small amount of native gG 2 to the peptide assay tended to increase the specificity. CONCLUSION: HSV gG and gD peptides show promise as type-specific and type-common HSV antigens. These peptides are more stable and reproducibly prepared than native or recombinant glycoproteins and may be considered for inclusion in future HSV serodiagnostic assays. PMID- 10382823 TI - Cytokine release in mononuclear cells of patients with Co-Cr hip prosthesis. AB - The aim of this study was the evaluation of the release of bone-resorbing cytokines in peripheral blood mononuclear cells (PBMC) of patients with aseptic loosening of Co-Cr hip prostheses. TNF-alpha, IL-6, and GM-CSF were measured in both unstimulated and PHA-stimulated PBMC, and in PBMC cultured in the presence of chromium and cobalt extracts. Blood samples from healthy donors were used as controls. Serum samples of both patients and healthy donors were tested to determine the concentration of metal ions. The proportion of lymphocyte, monocyte and lympocyte subpopulation in cultured PBMC did not differ in patients and the control group. In unstimulated PBMC the release of TNF was significantly higher in patients than in the control group, while IL-6 was significantly decreased and no change was observed for GM-CSF. When the PBMC were challenged with chromium extract, all the 'index of cytokine release' resulted higher in patients than in the control group; cobalt extract increased both the TNF and GM-CSF index, but not the index of IL-6 release. Metal concentrations in serum from patients were significantly increased and correlated with the TNF release in PBMC stimulated with both metal extract. Our results suggest that a CoCr-implant releases a large amount of metal ions which could mediate the priming or the renewal of a cell mediated hypersensitivity reaction. The prevalence of circulating lymphocytes responsible for the delayed hypersensitivity, namely Thl, would justify both the significant increase of TNF and the significant decrease of IL6 in unstimulated PBMC of patients, as well as the significant increase of the 'index of cytokine release' after the challenge with metal ions. PMID- 10382824 TI - Quantitative comparison of osteoconduction of porous, dense A-W glass-ceramic and hydroxyapatite granules (effects of granule and pore sizes). AB - The osteoconductive potentials of dense, small porous and large porous apatite- and wollastonite-containing glass-ceramic (A-W GC) granules of various sizes implanted in rat tibiae were evaluated quantitatively, by determining their affinity indices. The average affinity indices of all types of A-W GC were high. The dense A-W GC granules had the highest values (97.0+/-5.5%), followed by the large porous (87.1+/-8.4%) and then the small porous granules (79.0+/-8.4%). There were no significant differences among the osteoconductive potentials of the different sizes of each form of A-W GC granule. The osteoconductive potentials of four types of commercially available porous hydroxyapatite (HA) granules were compared with those of the small porous A-W GC granules, using the affinity index and the proportion of newly formed bone relative to that of the granules in the bone defect. The values of the former parameter for three types of HA and those of the latter for four were significantly inferior to those of A-W GC. The difference between the osteoconductive potentials of A-W GC and HA was considered to be related to the rate of surface apatite layer formation. PMID- 10382825 TI - An osteoconductive collagen/hyaluronate matrix for bone regeneration. AB - A new type of collagen-hyaluronate (COL/HA) matrix was synthesized by cross linking collagen fibers with modified hyaluronate polymers bearing active formyl groups. The resulting matrix is a three-dimensional scaffold consisting of interconnected pores with an average size of 40 microm and a high pore volume/surface area ratio. The covalent nature of the bond between the collagen fibers and the modified hyaluronate was demonstrated by extended elution with phosphate buffered saline and by extraction in increasing ionic gradients. The fraction of covalently bound hyaluronate in the matrix ranged from 5 to 25 w%. The total hyaluronate content of the COL/HA matrix affected both the in vitro non enzymatic and enzymatic degradation as well as the in vivo turnover. When implanted in cranial defects in rats, the COL/HA matrix demonstrated good biocompatibility and exhibited greater osteoconductive potential than matrices composed of either cross-linked collagen or cross-linked hyaluronate alone. PMID- 10382826 TI - In vivo evaluation of poly(L-lactic acid) porous conduits for peripheral nerve regeneration. AB - The present study provides in vivo trials of poly(L-lactic acid) (PLLA) as a porous biodegradable nerve conduit using a 10 mm sciatic nerve defect model in rats. The PLLA conduits, fabricated by an extrusion technique, had an inner diameter of 1.6 mm, an outer diameter of 3.2 mm, and a length of 12 mm. They were highly porous with an interconnected pore structure (of 83.5% porosity and 12.1 microm mean pore size). The conduits were interposed into the right sciatic nerve defect of Sprague Dawley rats using microsurgical techniques; nerve isografts served as controls. Walking track analysis was performed after conduit placement monthly through 16 weeks. At the conclusion of 6 and 16 weeks, sections from the isograft/conduit and distal nerve were harvested for histomorphometric analysis. The right gastrocnemius muscle was also harvested and its weight was determined. All conduits remained intact without breakage. Moreover, no conduit elongated during the 16 weeks of placement. Walking track analysis and gastrocnemius muscle weight demonstrated increasing regeneration over the 16 weeks in both the conduit and isograft control groups, with control values significantly greater. The nerve fiber density in the distal sciatic nerve for the PLLA conduits (0.16+/-0.07) was similar to that for the control isografts (0.19+/-0.05) at 16 weeks. The number of axons/mm2 in the distal sciatic nerve for the PLLA conduits was lower than that for the isografts (13 800+/-2500 vs. 10700+/-4700) at 16 weeks. The results for PLLA were significantly improved over those for 75:25 poly(DL-lactic-co glycolic acid) of a previous study and suggest that PLLA porous conduits may serve as a scaffold for peripheral nerve regeneration. PMID- 10382827 TI - Characterization of transient platelet contacts on a polyvinyl alcohol hydrogel by video microscopy. AB - Acridine orange labelled, washed human platelets were counted and tracked on polyvinyl alcohol (PVA), heparin-PVA and polyethylene (PE)-coated coverslips with a view to understand why transient contact on the PVA hydrogels lead to elevated platelet activation and consumption relative to polyethylene. Over the 4 min of initial contact that was studied, platelet adhesion was higher on PE than on PVA or heparin-PVA at both 40 and 200 s(-1), as expected, regardless of whether the surfaces were pre-treated with albumin or fibrinogen. Not all platelets appearing to make contact with the surface, actually attached. For example, less than 2% of the platelets contacting albumin pre-treated PVA (at 40 s(-1)) remained adherent at the end of the initial 60 s observation time, while the corresponding number for PE was greater than 9%. A greater fraction of the platelets remained adherent at the higher shear rate or with fibrinogen pre-treatment, but the difference between PVA and PE remained similar: for example, with fibrinogen pre-treatment at 200 s(-1), approximately 25% of the platelet contacts resulted in adhesion on PVA while 66% did so on PE. While net platelet adhesion was less for the hydrogels, than for PE, the total number of contacts (adherents + non-adherents) were more comparable and unexpectedly higher for albumin pre-treatment than for fibrinogen. Net platelet adhesion is but one component of the total platelet interaction with a material surface. Fluorescent video microscopy has been shown to be a useful, albeit not unequivocal, method for assessing the platelets that make contact with but do not adhere to a surface. reserved PMID- 10382828 TI - Apatite formation on PDMS-modified CaO-SiO2-TiO2 hybrids prepared by sol-gel process. AB - Dense and homogeneous monolithics of polydimethylsiloxane (PDMS)-modified CaO SiO2-TiO2 hybrids were successfully synthesized by sol-gel process. They were assumed to be composed of a silica and titania network incorporated with PDMS and the calcium ion ionically bonded to the network. Among them, the hybrids containing relatively larger amounts of the calcium in their surfaces formed an apatite on their surfaces within only 1 or 0.5 day in a simulated body fluid with ion concentrations nearly equal to those of human flood plasma. This indicates that they are highly bioactive. Organic-inorganic hybrids synthesized by a sol gel process usually exhibit high ductility, low elastic modulus and high mechanical strength. The present hybrids are, therefore, expected to be useful as a new kind of bone-repairing material, because of their high bioactivity and unique mechanical properties. PMID- 10382829 TI - Porous chitosan scaffolds for tissue engineering. AB - The wide array of tissue engineering applications exacerbates the need for biodegradable materials with broad potential. Chitosan, the partially deacetylated derivative of chitin, may be one such material. In this study, we examined the use of chitosan for formation of porous scaffolds of controlled microstructure in several tissue-relevant geometries. Porous chitosan materials were prepared by controlled freezing and lyophilization of chitosan solutions and gels. The materials were characterized via light and scanning electron microscopy as well as tensile testing. The scaffolds formed included porous membranes, blocks, tubes and beads. Mean pore diameters could be controlled within the range 1-250 microm, by varying the freezing conditions. Freshly lyophilized chitosan scaffolds could be treated with glycosaminoglycans to form ionic complex materials which retained the original pore structure. Chitosan scaffolds could be rehydrated via an ethanol series to avoid the stiffening caused by rehydration in basic solutions. Hydrated porous chitosan membranes were at least twice as extensible as non-porous chitosan membranes, but their elastic moduli and tensile strengths were about tenfold lower than non-porous controls. The methods and structures described here provide a starting point for the design and fabrication of a family of polysaccharide based scaffold materials with potentially broad applicability. PMID- 10382830 TI - Mechanical loading of bovine pericardium accelerates enzymatic degradation. AB - Bioprosthetic heart valves fail as the result of two simultaneous processes: structural deterioration and calcification. Leaflet deterioration and perforation have been correlated with regions of highest stress in the tissue. The failures have long been assumed to be due to simple mechanical fatigue of the collagen fibre architecture; however, we have hypothesized that local stresses-and particularly dynamic stresses-accelerate local proteolysis, leading to tissue failure. This study addresses that hypothesis. Using a novel, custom-built microtensile culture system, strips of bovine pericardium were subjected to static and dynamic loads while being exposed to solutions of microbial collagenase or trypsin (a non-specific proteolytic enzyme). The time to extend to 30% strain (defined here as time to failure) was recorded. After failure, the percentage of collagen solubilized was calculated based on the amount of hydroxyproline present in solution. All data were analyzed by analysis of variance (ANOVA). In collagenase, exposure to static load significantly decreased the time to failure (P < 0.002) due to increased mean rate of collagen solubilization. Importantly, specimens exposed to collagenase and dynamic load failed faster than those exposed to collagenase under the same average static load (P = 0.02). In trypsin, by contrast, static load never led to failure and produced only minimal degradation. Under dynamic load, however, specimens exposed to collagenase, trypsin, and even Tris/CaCl2 buffer solution, all failed. Only samples exposed to Hanks' physiological solution did not fail. Failure of the specimens exposed to trypsin and Tris/CaCl2 suggests that the non-collagenous components and the calcium-dependent proteolytic enzymes present in pericardial tissue may play roles in the pathogenesis of bioprosthetic heart valve degeneration. PMID- 10382831 TI - Best practice--ongoing polemics. AB - Current treatment strategies and disease management programs for hyperlipidemia employ a range of lipid-lowering drugs. Results from early lipid-lowering trials using diet, fibrates, niacin and other classes of drug showed that lowering plasma cholesterol can significantly reduce the risk of developing ischemic cardiovascular events. The landmark statin trials have clearly demonstrated the benefits of lipid-lowering therapy in coronary heart disease (CHD) prevention and unlike early lipid-lowering studies, a reduction in mortality may become evident with statin therapy during the first year of treatment. The number of successful lipid-intervention trials continues to increase and evidence is accumulating that lipid modification can also reduce the risk of cardiovascular events among individuals with only modest degrees of blood-lipid abnormalities. With increasingly powerful drugs to modify blood lipids, the potential levels at which to initiate treatment and the appropriate target levels are rapidly changing and debate surrounds the question of where the line to initiate treatment should be drawn. The relative lack of major adverse events with statin therapy means that the level of CHD risk at which clinical benefit occurs cannot be determined by the degree of risk at which benefit exceeds adverse events. Therefore, patients with only moderately raised cholesterol levels can be treated because statin treatment is well tolerated. One of the most important aspects of the statin trials is the finding that clinical events, such as death and disability due to coronary artery disease, may be preventable or limited in a significant number of patients if they receive aggressive therapy. Current goals for cholesterol levels in patients with established CHD are rarely achieved with non-aggressive treatment; however, with aggressive lipid lowering statins can achieve these goals in a safe and effective manner. PMID- 10382832 TI - Guidelines--a missed opportunity. AB - Progress is slow in the integration of coronary heart disease prevention into daily clinical practice by cardiologists and other physicians working in cardiology, internal medicine and primary healthcare. This is not due to a lack of professional recommendations on coronary prevention. As new knowledge has become available, new generations of recommendations have been published. In surveys of secondary prevention practice, risk factor recording and management falls short of these recommendations and there is considerable potential to further reduce the risk of another major ischemic event in patients with established coronary disease. The most recent recommendations from the Joint Task Force of the European Society of Cardiology, European Atherosclerosis Society and European Society of Hypertension reinforce the importance of secondary prevention and how they can be extended to primary prevention by targeting high-risk individuals in the general population. It is hoped that development of guidelines at a national level will ensure a common approach to coronary heart disease prevention throughout Europe. PMID- 10382833 TI - Medical education--communicating best practice. AB - There is unequivocal evidence that reduction of modifiable risk factors for coronary heart disease (CHD), such as elevated serum cholesterol, high blood pressure and smoking, decreases cardiovascular mortality. However, levels of intervention appear to be poor and there is often little evidence of a combined multiple risk factor approach to intervention, which is likely to be the optimal way to lower a patients overall risk of developing CHD. Attention has recently focused on bridging the gap between knowledge in the field of preventive cardiology and its application in everyday clinical practice. This can only be accomplished by the development of uncomplicated, practical guidelines that are evidenced-based and that provide specific targets for risk factors and indications for drug therapy. PMID- 10382834 TI - The future of best practice. AB - Evidence from epidemiological and clinical studies continues to improve our understanding of the pathogenesis of coronary heart disease (CHD). However, despite major advances in the development of diagnostic methods and effective treatment, CHD remains the leading cause of mortality in the Western world. That cholesterol lowering is of major importance in lowering the risk of developing coronary artery disease (CAD) is now an accepted principle in medicine. Most patients with CAD will require drug therapy to achieve target lipid levels. Subgroup analyses of data from the landmark statin trials show that this benefit is seen in all patient groups: male and female, older and younger patients, diabetics and non-diabetics, and patients with and without myocardial revascularization. Recent evidence suggests that the extent to which cholesterol is lowered is also important and that the attainment of reduced low-density lipoprotein cholesterol levels is associated with greater reductions in the risk of cardiovascular events. Unfortunately, data from studies to date do not provide a definitive answer to the question of whether there is a benefit in lowering cholesterol to very low levels. PMID- 10382835 TI - Different brain radioactivity curves in a PET study with [11C]beta-CIT labelled in two different positions. AB - The cocaine congener 2beta-carbomethoxy-3beta-(4'-iodophenyl)tropane (beta-CIT) has a chemical structure that enables labelling with carbon-11 either by N methylation or by O-methylation. The regional brain uptake of [N-methyl-11C]beta CIT and [O-methyl-11C]beta-CIT was compared in cynomolgus monkeys using positron emission tomography (PET). The striatal uptake of radioactivity after intravenous injection of [O-methyl-11C]beta-CIT reached a plateau at 30-40 min, whereas the uptake of [N-methyl-11C]beta-CIT increased continuously during the time of the PET measurement. Two of the putative labelled metabolites, [N-methyl-11C]beta-CIT acid and [O-methyl-11C]nor-beta-CIT were prepared and examined with PET to investigate if they may enter the brain and thus add to the radioactivity uptake obtained with [11C]beta-CIT. Less than 0.4% of injected [N-methyl-11C]beta-CIT acid entered the brain whereas 5-6% of [O-methyl-11C]nor-beta-CIT entered the brain and accumulated in the striatum and in the thalamus. The fraction of [O methyl-11C]nor-beta-CIT obtained in plasma after intravenous injection of [O methyl-11C]nor-beta-CIT, however, never exceeded 3%. Consequently, the formation of [N-methyl-11C]beta-CIT-acid and [O-methyl-11C]nor-beta-CIT cannot be the explanation for the different time-activity curves in the monkey brain demonstrated with [11C]beta-CIT labelled in two different positions. An unidentified labelled lipophilic metabolite, detected in monkey plasma after injection of [O-methyl-11C]beta-CIT, remains as the only possible explanation for the differences between [N-methyl-11C]beta-CIT and [O-methyl-11C]beta-CIT. PMID- 10382836 TI - High purity production and potential applications of copper-60 and copper-61. AB - Previously we described the high yield production of 64Cu using a target system designed specifically for low energy, biomedical cyclotrons. In this study, the use of this target system for the production of 60Cu and 61Cu is described and the utility of these isotopes in the labeling of biomolecules for tumor and hypoxia imaging is demonstrated. 60Cu and 61Cu were produced by the 60Ni(p,n)60Cu, 61Ni(p,n)61Cu, and 60Ni(d,n)61Cu nuclear reactions. The nickel target (>99% enriched or natural nickel) was plated onto a gold disk as described previously (54-225 microm thickness) and irradiated (14.7 MeV proton beam and 8.1 MeV deuteron beam). The copper isotopes were separated from the nickel via ion exchange chromatography and the radioisotopic purity was assessed by gamma spectroscopy. Yields of up to 865 mCi of 60Cu have been achieved using enriched 60Ni. 61Cu has been produced with a maximum yield of 144 mCi using enriched 61Ni and 72 mCi using enriched 60Ni. Specific activities (using enriched material) ranged from 80 to 300 mCi/microg Cu for 60Cu and from 20 to 81 mCi/microg Cu for 61Cu. Bombardments of natural Ni targets were performed using both protons and deuterons. Yields and radioisotopic impurities were determined and compared with that for enriched materials. 60Cu was used to radiolabel diacetyl-bis(N4 methylthiosemicarbazone), ATSM. 60Cu-ATSM was injected into rats that had an occluded left anterior descending coronary artery. Uptake of 60Cu-ATSM in the hypoxic region of the heart was visualized clearly using autoradiography. In addition, 60Cu-ATSM was injected into dogs and excellent images of the heart and heart walls were obtained using positron emission tomography (PET). 61Cu was labeled to 1,4,8,11-tetraazacyclotetradecane-N,N',N",N"'-tetraacetic acid octreotide (TETA-octreotide) and the PET images of tumor-bearing rats were obtained up to 2 h postinjection. After decay of the 61Cu, the same rat was injected with 64Cu-TETA-octreotide and the images were compared. The tumor images obtained using 61Cu were found to be superior to those using 64Cu as predicted based on the larger abundance of positrons emitted by 61Cu vs. 64Cu. PMID- 10382838 TI - [18F]Fluoro-beta-fluoromethylene-m-tyrosine derivatives show stereo, geometrical, and regio specificities as in vivo central dopaminergic probes in monkeys. AB - Stereo (D and L), geometrical (E and Z), and regiospecific (2-, 4-, and 6 [18F]fluoro) analogs of beta-fluoromethylene-m-tyrosine (FMMT) have been investigated in adult vervet monkeys (Cercopithecus aethiops sabaeus, n = 12) in vivo with positron emission tomography (PET). Brain transport through the blood brain barrier and central aromatic amino acid decarboxylase (AAAD)-mediated decarboxylation rates were established. Results show strict structural dependency of the kinetic behavior of radiofluorinated FMMT analogs, with the E-isomer exhibiting a higher specificity over the (Z) geometrical counterpart for central dopaminergic structures. The 6-[18F]fluoro substituted L-(E)-FMMT was also favored over the 2- and 4-[18F]fluorosubstituted isomers in terms of their ability to localize in the same brain areas. The role of PET in drug development is also exemplified in this work. PMID- 10382837 TI - Synthesis of stereo (R and S) and geometric (E and Z) isomers of [18F]fluoro-beta fluoromethylene-m-tyrosine derivatives: in vivo probes of central dopaminergic function. AB - Fluorination of pure R and S enantiomers of (E)-beta-fluoromethylene-m-tyrosine [(E)-FMMT] and its racemic geometric isomer, (Z)-beta-fluoromethylene-m-tyrosine [(Z)-FMMT] with [18F]acetyl hypofluorite ([18F]AcOF) gave a mixture of aromatic ring fluorinated products and a pair of diastereomeric products of addition across the exocyclic double bond. Semipreparative high performance liquid chromatography (HPLC) enabled a complete separation and isolation of these products, namely, 6-[18F]fluoro, 2-[18F]fluoro, and 2,6-[18F]difluoro (E)-FMMT and (Z)-FMMT derivatives. No attempt was made to isolate the individual components of the addition product. Pure racemic 4-[18F]fluoro-(E)-beta fluoromethylene-m-tyrosine was also synthesized from a substituted (E)-FMMT precursor involving a fluorodestannylation reaction with [18F]F2. The availability of stereo (R and S) isomers of 6-[18F]fluoro and 2-[18F]fluoro (E) FMMT and those of the racemic (Z)-FMMT along with 4-[18F]fluoro-(E)-beta fluoromethylene-m-tyrosine would now enable a systematic investigation of the central monoamine oxidase/aromatic amino acid decarboxylase enzyme system with positron emission tomography. PMID- 10382839 TI - Evaluation of 9-[(3-18F-fluoro-1-hydroxy-2-propoxy)methyl]guanine ([18F]-FHPG) in vitro and in vivo as a probe for PET imaging of gene incorporation and expression in tumors. AB - Preparation of 9-[(3-18F-fluoro-1-hydroxy-2-propoxy)methyl]-guanine ([18F]-FHPG) for clinical use, and its evaluation as a positron emission tomography (PET) imaging agent for gene incorporation and expression in tumors are reported. In vitro studies in human colon cancer cells, HT-29, transduced with the retroviral vector G1Tk1SvNa and nontransduced (wild type) showed 4, 8, 12, and 15 times higher uptake of the probe in 1, 3, 5, and 7 h, respectively, in transduced cells compared with the controls. In vivo studies in tumor-bearing nude mice demonstrated that the tumor uptake of the radiotracer is three and six-fold higher in 2 and 5 h, respectively, in transduced cells compared with the control cells. These results suggest that [18F]-FHPG is a potential in vivo PET imaging agent for monitoring gene incorporation and expression in gene therapy of cancer. PMID- 10382841 TI - Transfer of 131I and fluoresceinyl sialic acid derivatives into the oligosaccharide chains of IgG: a new method for site-specific labeling of antibodies. AB - Biochemical modifications of IgG can help to avoid damages caused by oxidation or reduction. Terminal groups of the saccharide structures, located in the Fc portion of IgG molecules, were modified by enzymatic reactions. IgG was pretreated with sialidase, to cleave bound sialic acid, and with galactosyltransferase, to increase the number of acceptor sites for transfer reactions. Afterward, modified sialic acid derivatives were transferred enzymatically into the oligosaccharide chains of IgG. Labeling was possible with sialic acids modified in either position 5 or position 9. The usefulness of this method was demonstrated for radioactive and fluoresceinylated reagents, with yields up to 90% in 1 h. Immunological investigations have shown no influence on the immunoreactivity by the described modification of saccharide structures. PMID- 10382840 TI - Synthesis, in vitro pharmacologic characterization, and preclinical evaluation of N-[2-(1'-piperidinyl)ethyl]-3-[125I]iodo-4-methoxybenzamide (P[125I]MBA) for imaging breast cancer. AB - The goal of this study was to investigate the potential use of a radioiodinated benzamide, N-[2-(1'-piperidinyl)ethyl]-3-iodo[125I]-4-methoxybenzamide (P[125I]MBA), a sigma receptor binding radioligand for imaging breast cancer. The chemical and radiochemical syntheses of PIMBA are described. The pharmacological evaluation of PIMBA was carried out for sigma-1 and sigma-2 receptor sites. The in vivo pharmacokinetics of the radioiodinated benzamide were determined in rats and comparison of P[125I]MBA with Tc-99m sestamibi were made in a rat mammary tumor model. Sigma-1 affinity (Ki) for PIMBA in guinea pig brain membranes using [3H](+)pentazocine was found to be 11.82 +/- 0.68 nM, whereas sigma-2 affinity in rat liver using [3H]DTG (1,3-o-di-tolylguanidine) was 206 +/- 11 nM. Sites in guinea pig brain membranes labeled by P[125I]MBA showed high affinity for haloperidol, (+)-pentazocine, BD1008, and PIMBA (Ki = 4.87 +/- 1.49, 8.81 +/- 1.97, 0.057 +/- 0.005, 46.9 +/- 1.8 nM, respectively). Competition binding studies were carried out in human ductal breast carcinoma cells (T47D). A dose dependent inhibition of specific binding was observed with several sigma ligands. Ki values for the inhibition of P[125I]MBA binding in T47D cells for haloperidol, N-[2-(1'-piperidinyl)]ethyl]4-iodobenzamide (IPAB), N-(N-benzylpiperidin-4-yl)-4 iodobenzamide (4-IBP), and PIMBA were found to be 1.30 +/- 0.07, 13 +/- 1.5, 5.19 +/- 2.3, 1.06 +/- 0.5 nM, respectively. The in vitro binding data in guinea pig brain membranes and breast cancer cells confirmed binding to sigma sites. The saturation binding of P[125I]MBA in T47D cells as studied by Scatchard analysis showed saturable binding, with a Kd = 94 +/- 7 nM and a Bmax = 2035 +/- 305 fmol/mg of proteins. Biodistribution studies in Sprague-Dawley rats showed a rapid clearance of P[125I]MBA from the normal organs. The potential of PIMBA in imaging breast cancer was evaluated in Lewis rats bearing syngeneic RMT breast cancers, a cancer that closely mimics human breast cancer histology. At 1 h postinjection, tumor uptake for P[125I]MBA and Tc-99m sestamibi were found to be 0.35 +/- 0.01 and 0.32 +/- 0.01% injected dose/organ (%ID/g), respectively. The %ID/g for liver, kidneys, and heart were 2, 11, and 20 times lower, respectively, for P[125I]MBA as compared with Tc-sestamibi. Slightly higher uptake of P[125I]MBA in tumors (than Tc-sestamibi) and a low nontarget organ uptake warrants further studies of this and other sigma receptor ligands for their use as breast cancer imaging agents. PMID- 10382842 TI - 99m-Technetium-labelled peptide-HYNIC conjugates: effects of lipophilicity and stability on biodistribution. AB - The aim of this study was to explore the effects of lipophilicity and stability on the biodistribution of 99mTc labelled peptides through the use of different co ligands. 6-Hydrazinopyridine-3-carboxylic acid (HYNIC) was coupled to the somatostatin analogue RC160 and radiolabelled using a range of ethylendiaminediacetic acid (EDDA) and ethylenediaminetetraacetic acid (EDTA) derivatives as well as tricine and pyridine/tricine as co-ligands. After labelling with technetium-99m, chromatographic, stability, protein-binding, and rat biodistribution studies were performed. For most co-ligands, biodistribution correlated well with in vitro properties. Lipophilic substitution on EDDA resulted in higher protein binding, increased liver uptake, and intestinal excretion. Stabilisation of tricine with pyridines reduced blood levels and lowered liver uptake. EDTA derivatives showed high instability in vitro and in vivo. PMID- 10382844 TI - Astatine-211 labeling of internalizing anti-EGFRvIII monoclonal antibody using N succinimidyl 5-[211At]astato-3-pyridinecarboxylate. AB - Monoclonal antibodies (MAbs) such as the anti-epidermal growth factor variant III (EGFRvIII) MAb L8A4 are rapidly internalized, which can lead to rapid loss of radioactivity from the tumor cell. The aim of this study was to evaluate the potential utility of N-succinimidyl 5-[211At]astato-3-pyridinecarboxylate ([211At]SAPC) for labeling murine L8A4 with 211At. SAPC was synthesized by astatodestannylation of N-succinimidyl 5-tri-n-butylstannyl 3-pyridinecarboxylate and then coupled to L8A4 in approximately 50% yield. The affinity and immunoreactive fraction for 211At-labeled L8A4 were comparable to those obtained when the MAb was labeled with 131I via N-succinimidyl 5-[131I]iodo-3 pyridinecarboxylate (SIPC). Paired-label comparisons of the 211At- and 131I labeled MAbs demonstrated similar internalization and catabolism by EGFRvIII positive cells in vitro, and with the exception of the stomach, similar tissue distribution in athymic mice with EGFRvIII-expressing U87MGdeltaEGFR xenografts. These results suggest that SAPC may be a useful reagent for labeling L8A4, and possibly other internalizing proteins, with 211At. PMID- 10382843 TI - Synthesis, purification, and in vitro stability of 211At- and 125I-labeled amidobisphosphonates. AB - A method is described for preparing 211At- and radioiodinated amidobisphosphonates. The active esters N-succinimidyl 3-(tri-methylstannyl) benzoate (ATE) and N-succinimidyl 5-(tri-methylstannyl)-3-pyridinecarboxylate (SPC) were used as precursors. The isolated and purified radiolabeled intermediates were coupled to 3-amino-1-hydroxypropylidene-1,1-bisphosphonate (APB) in high yields ranging from 60% to 97%. The lipophilicity of the compounds was found to depend on the nature of the labeled template and the halogen. High in vitro stability in mouse, fetal calf, and human serum was documented by high performance liquid chromatography. PMID- 10382845 TI - In vivo dynamic distribution of 131I-glucagon-like peptide-1 (7-36) amide in the rat studied by gamma camera. AB - The in vivo distribution of glucagon-like peptide-1 (7-36) amide (GLP-1) was studied in a rat model using radiolabeled GLP-1 (131I-GLP-1) depicted by a gamma camera. The dynamic scan showed a rapid clearance from the blood circulation after an intravenous (i.v.) injection of 131I-GLP-1. After 10 min, the major part of the radioactivity was accumulated in the kidneys, whereas about 9% (of the blood value) was found in the brain. The pharmacokinetic study using 125I-GLP-1 demonstrated a rapid elimination from plasma, with a half-life of 3.3 +/- 0.6 min, a clearance of 117 +/- 15 mL/min, and a distribution volume of 557 +/- 61 mL. The elimination half-lives for the intact 125I-GLP-1 in lungs and kidneys were determined to 3.7 and 3.9 min, respectively. The metabolite GLP-1 (9-36) amide was followed in blood, lung, and kidney. All other organs assumed to contain low molecular weight fragments of GLP-1. The present study suggest that GLP-1 and/or its labeled metabolites cross the blood-brain barrier. Also the kidney plays an essential role in GLP-1 elimination after an i.v. administration, which can be of clinical interest especially in patients with kidney insufficiency who are treated with GLP-1. PMID- 10382846 TI - Metal complexes of bleomycin: evaluation of [Rh-105]-bleomycin for use in targeted radiotherapy. AB - Bleomycin has been used as a carrier for several radioisotopes; however, its potential for clinical use has been limited either by the in vivo stability of the complexes or the half-life of the isotope used. The chemical, biological, and radiological properties of 105Rhodium appear to make it an ideal choice for targeted radiotherapy. The synthesis and purification of a hereto unreported 105Rhodium-bleomycin (105Rh-BLM) complex is described. The stability of this complex in plasma is sufficient to allow targeted delivery of the radioisotope. 57Cobalt-bleomycin was studied under identical conditions for comparative purposes. The suitability of 105Rh-BLM for targeted therapy, which appears to be limited by the renal clearance of this agent, is discussed. PMID- 10382847 TI - High yield synthesis of [11C]-acetone through selective quenching of methyl lithium. AB - Carbon-11 labeled acetone is a useful radiosynthetic precursor. Previously, strict control of no-carrier-added stoichiometry was required to prepare it from reaction of CO2 and methyl lithium. However, excess methyl lithium may be selectively quenched to avoid reaction with nascent acetone to give tert-butanol. We report a simple pKa-based strategy to sequentially and selectively quench MeLi and acetone to give yields of up to 100% acetone even in the presence of a large excess of MeLi. The method gives good yields of acetone under conditions that previously precluded its synthesis. PMID- 10382848 TI - Mechanism of nitrogen-13-labeled ammonia formation in a cryogenic water target. AB - Methods for producing N-13 ammonia via the 16O(p,alpha)13N nuclear reaction utilizing a cryogenic target have been investigated. These targets included frozen carbon dioxide and pure frozen water. Results from these targets were compared with the more traditional liquid water target with and without additives. A very dramatic difference was found between the pure water target in the frozen state when compared with the liquid state. When frozen, more than 95% of the nitrogen-13 activity is in the chemical form of ammonia at all radiation doses. In contrast, the liquid water target yielded predominately nitrates and nitrites at high radiation doses. When frozen carbon dioxide was irradiated under these conditions, more than 95% of the nitrogen-13 activity was in the form of nitrate and nitrite. The nitrogen oxides remained on the surface of the target and could be easily removed from the surface with pure water. The wash solution was converted to [13N]ammonia using the DeVarda's alloy method for reduction. It was determined that levels of [13N]ammonia sufficient for diagnostic medical procedures could be produced directly using the frozen water targets or from frozen carbon dioxide with a wet chemical reduction. These results have significance particularly in the design of targetry for low-energy, high-beam current accelerators, because targets of this design can be used with either no vacuum isolation window or a very thin window. The substitution of carbon-13 enriched carbon dioxide for natural carbon dioxide gives access to the 13C(p,n)13N nuclear reaction, which allows protons energies as low as 6 MeV to be used to produce useable quantities of N-13 ammonia. The mechanism of these reactions has been explored to determine why there are such dramatic differences in the product distribution between the frozen state and the liquid or gaseous state. PMID- 10382850 TI - Effect of reaction conditions on preparations of rhenium-188 hydroxyethylidene diphosphonate complexes. AB - Rhenium-186 (Re-186) hydroxyethylidene diphosphonate (HEDP) has been shown to localize in metastatic foci within bone in a manner similar to Tc-99m bone seeking agents. Usually, in the preparation of diagnostic Tc-99m radiopharmaceuticals, the concentration of Tc is at trace level (10(-8) M). However, large amounts of carrier are included in the preparation of Re-186 radiopharmaceuticals (10(-4) M), which may significantly affect the preparation of Re-HEDP. In this study, Re-188 was used as an Re tracer. The effects of pH and concentrations of Re carrier on the preparation of Re-HEDP were investigated. Re 188-Sn-HEDP was prepared by reconstitution of a kit of lyophilized HEDP mixture, and tin chloride with a radioactive solution of perrhenate in saline. The total concentration of Re present in this work ranged from 10(-8) to 10(-3) M. The results showed that high labeling efficiency was obtained for each preparation. Although the chemical behaviors of the Re-188 HEDP complexes, with and without carrier, were similar, the biodistribution patterns of carrier free Re-188 HEDP in rats were found to differ from the biodistribution patterns of carrier-added Re-188 HEDP. PMID- 10382849 TI - Investigations of acetonitrile solvent cluster formation in supercritical carbon dioxide, and its impact on microscale syntheses of carbon-11-labeled radiotracers for PET. AB - A new strategy has been developed for synthesizing positron emission tomography (PET) radiotracers using [11C]methyl iodide. This strategy relies on the ability of organic co-solvents to cluster within mixtures of supercritical fluids resulting in localized regions of high density which can serve as microscopic pockets for reaction. We've shown that acetonitrile will cluster about dilute solutes when mixtures of this co-solvent with carbon dioxide are forced to behave as a homogeneous fluid at the critical point. We applied this strategy in a systematic investigation of the conditions for optimized reaction between methyl iodide and L-alpha-methyl-N-2-propynyl phenethylamine (nordeprenyl) to yield L deprenyl. Variables such as temperature, ultraviolet light exposure, co-solvent concentration, system pressure, and methyl iodide concentration were explored. The synthesis of radioactive [11C]-L-deprenyl using no-carrier-added concentrations of [11C]methyl iodide was also tested. Results showed that greater than 90% radiochemical yield of the desired product could be attained using 40 times less labeling substrate than in conventional PET tracer syntheses. PMID- 10382852 TI - Convenient gas phase bromination of [11C]methane and production of [11C]methyl triflate. PMID- 10382851 TI - An improved kit formulation of a dopamine transporter imaging agent: [Tc 99m]TRODAT-1. AB - Recently, [Tc-99m]TRODAT-1, the first Tc-99m-labeled tracer for imaging CNS dopamine transporters in humans, was reported. This tracer displayed excellent specific binding to dopamine transporters in the basal ganglia region of the brain, thus it is potentially useful for the diagnosis of deficit of dopamine transporters in neurodegenerative diseases, such as Parkinson's disease. Preparation of [Tc-99m]TRODAT-1 was previously achieved by a multistep kit formulation. It is highly desirable to further improve the preparation by developing a simplified one-vial formulation with a reduced amount of TRODAT-1 ligand for routine clinical use. To achieve this goal, a series of studies to optimize labeling efficiency by varying a combination of factors (amount of free ligand, reaction reagents, and reaction pH) was carried out. [Tc-99m]TRODAT-1 prepared by this new kit formulation was evaluated by assessing the brain uptake and target (striatum) versus nontarget (cerebellum) ratios in rats. Appropriate amounts of various ingredients for a one-vial kit formulation providing > or =90% radiolabeling yields were identified. The most consistent and reliable formulation contained 10 microg of TRODAT-1 (a reduction of free ligand from 200 microg to 10 microg), 32 microg of SnCl2, 10 mg of sodium glucoheptonate, and 840 microg of disodium EDTA in one vial as a lyophilized kit. It is feasible to reconstitute the vial with [Tc-99m]pertechnetate (0.5-2 mL, < or =1110 MBq, 30 mCi), resulting in a final solution with a pH value of 4.5-5.0. [Tc-99m]TRODAT-1, prepared by this new kit, was stable at room temperature for 6 h. Biodistribution studies of this agent in rats with the new formulation showed similar regional brain distribution as compared with those obtained with the previous preparation (high striatum-to-cerebellum ratio). In conclusion, using this lyophilized one vial kit formulation, [Tc-99m]TRODAT-1 can be prepared with greater than 90% radiochemical purity. This simplified kit will significantly improve the reliability of preparation of this agent for routine clinical use. PMID- 10382853 TI - Automated production of 16alpha-[18F]fluoroestradiol for breast cancer imaging. PMID- 10382854 TI - Pharmacological applications of inorganic complexes. AB - Inorganic complexes have long been utilized for many therapeutic purposes. They were used or tried, perhaps because of the general notion that inorganic compounds (e.g., metal complexes) are toxic and a controlled use of such a compound may suppress some biological process. In this review, we briefly outline the properties of several selected groups of inorganic complexes and how they can affect biological systems and contribute to human pathologies. PMID- 10382855 TI - Mechanism of inhibition of the Na current by tocainide in guinea-pig isolated ventricular cells. AB - Tocainide blocked the Na current (I(Na)) in ventricular myocytes in either a tonic or a phasic block manner, having a higher affinity for the inactivated state (K(di) = 18 microM) than for the rested state (Kd(rest) = 606 microM). The degree of phasic block was enhanced and the onset of phasic block was faster with an increase in pulse duration as well as at less-negative holding potential (HP). The recovery-time constant from the phasic block of I(Na) induced by tocainide was independent of either the HP or the removal of fast inactivation. After removal of fast inactivation, tocainide showed the pulse-duration dependency of phasic block but not the voltage dependency. These results suggest that tocainide could bind to the inactivated-state receptor through the hydrophilic pathway and leave the receptor through the hydrophobic pathway, producing the tonic and phasic block of I(Na). PMID- 10382856 TI - Sympathomimetic effects of Parquetina nigrescens (Periplocaceae) extract in isolated portal vein smooth muscle. AB - The purpose of this study was to examine the mechanisms underlying the pharmacological effects of the extract of Parquetina nigrescens (Expar) on vascular smooth muscle contractility. To evaluate the Expar effect, the contractile activity of portal veins isolated from Wistar Kyoto rats was isometrically recorded. Isolated portal vein preparations developed rhythmic and spontaneous contractile activity. Expar increased the contractile response of the portal vein preparations in a dose-dependent manner. The maximal effect of the dose-response curve for Expar was prevented by the alpha1-adrenergic blocking agent prazosin at 10 nM and 30 nM concentration dependently. The contractile responses of the muscle to Expar were partly blocked after chemical sympathectomy of the preparations with 6-hydroxydopamine, and those obtained in the same conditions with tyramine were completely abolished, whereas responses to noradrenaline were unaffected by the 6-hydroxydopamine pretreatment. It is concluded that Expar contains principles, which can be characterized as direct and indirect sympathomimetic. PMID- 10382857 TI - Atypical beta-adrenoceptors of rat thoracic aorta. AB - The possible existence of atypical beta-adrenoceptors in vascular smooth muscle of rat isolated thoracic aorta was investigated. Isoprenaline (10(-8)-10(-4) M) produced concentration-dependent relaxation of phenylephrine (10(-5) M) precontracted rings of endothelium-denuded rat aorta in vitro. Isoprenaline induced relaxation was resistant to blockade by atenolol (10(-6) M). But, propranolol (2 x 10(-7) M) caused a non-competitive inhibition and SR 59230A (6.6 x 10(-6) M), a beta3-adrenoceptor selective antagonist, failed to produce additional antagonism in presence of propranolol. BRL 37344 (10(-8)-10(-4) M), a beta3-selective agonist, did not relax ring segments precontracted with phenylephrine (10(-5) M) in the absence of endothelium. The non-conventional partial agonist (-)-cyanopindolol (5 x 10(-6)-10(-4) M) induced a marked relaxation in phenylephrine (10(-5)M) precontracted aortic rings without endothelium. This vasodilation was resistant to blockade by propranolol (2 x 10( 7) M) and SR 59230A (10(-5) M). Salbutamol (10(-8)-10(-4) M) produced concentration-dependent relaxation in isolated endothelium-denuded aortic rings precontracted with phenylephrine (10(-5) M). Propranolol (2 x 10(-7) M), but not atenolol (10(-6) M), inhibited this relaxant response. It is concluded that in endothelium-denuded thoracic aorta, salbutamol acts through beta2-adrenoceptors whereas isoprenaline seems to activate both beta2-adrenoceptors and an atypical beta-adrenergic receptor. This atypical beta-adrenoceptor is distinct from putative beta3-adrenoceptor and maybe resembles the reported fourth cardiac beta adrenoceptor. PMID- 10382858 TI - Source(s) of activator calcium for noradrenaline-induced vasoconstriction in the perfused rabbit isolated ovarian vascular bed: a role for tyrosine kinase. AB - The effects of Ca2+ withdrawal and agents affecting Ca2+ translocation on alpha1 adrenoceptor-mediated vasoconstrictor responses in the perfused rabbit ovarian vascular bed were studied. Noradrenaline-induced vasoconstriction was lost in a Ca(2+)-free Krebs' solution, and the rate of loss of the response was accelerated by EGTA (2 mM). Noradrenaline-induced vasoconstriction and SDZ NVI 085-induced vasoconstriction were concentration-dependently inhibited by verapamil and nifedipine. These agents were, however, more effective against KCl-induced responses. Cyclopiazonic acid, an intracellular Ca(2+) depletor, concentration dependently inhibited noradrenaline-induced responses and abolished the response in Ca(2+)-free Krebs' solution. GF 109203X and polymyxin B, inhibitors of protein kinase C (PKC), had no significant effect on noradrenaline-induced responses. Tyrosine kinase inhibitors, genistein and erbstatin, inhibited noradrenaline induced vasoconstriction in the perfused rabbit ovarian vascular bed. The results would suggest that both extracellular Ca2+ and intracellular Ca2+ participate in noradrenaline-induced vasoconstrictor responses in the perfused rabbit ovarian vascular bed. The results would also suggest that tyrosine kinase and not protein kinase C activation has a role in such effects. PMID- 10382859 TI - Role of nitric oxide in rat coronary flow regulation during respiratory and metabolic acidosis. AB - A rat Langendorff heart preparation, perfused at constant pressure, was used to evaluate the role of nitric oxide in the increases in coronary vessel flow during hypercapnic and metabolic acidosis. Prior administration of the nitric oxide synthase (NOS) inhibitor N6-nitro-L-arginine methyl ester (100 microM) significantly reduced the basal, resting, rate of coronary flow but did not attenuate the increases in flow during brief (2-min) periods of perfusion with acidotic solutions. These results suggest that nitric oxide is not a significant contributor to rat heart coronary flow regulation during respiratory or metabolic acidosis. PMID- 10382860 TI - Use of brain slices in the study of pathogenic role of inducible nitric oxide synthase in cerebral ischemia-reperfusion. AB - We have recently demonstrated that inducible nitric oxide synthase (iNOS) is expressed in rat forebrain slices exposed to oxygen and glucose deprivation (OGD). Now, we have found that the expression of iNOS after OGD is time-dependent since 20 min of OGD produces the appearance of iNOS at earlier times than 10 min of OGD. OGD also causes neurotoxicity in this model, as revealed by the increase in excitatory amino acid, neuron specific enolase and lactate dehydrogenase (LDH) efflux to the incubation solution. Finally, the administration of the NMDA receptor antagonist MK-801 (100 nM) inhibits both the expression of iNOS and the release of LDH. Our findings demonstrate that this method may be considered an useful in vitro model of ischemia-reperfusion to determine the therapeutic role of neuroprotective tools. PMID- 10382861 TI - Modulation of adrenergic contraction of dog pulmonary arteries by nitric oxide and prostacyclin. AB - The aim of this work was to investigate the influence of endothelium-derived nitric oxide and prostaglandins on the contractile responses of isolated dog pulmonary arteries to electrical field stimulation and noradrenaline. Electrical field stimulation (1-8 Hz, 20 v, 0.25 ms duration, for 30 s) produced frequency dependent contractions that were abolished by tetrodotoxin, guanethidine and, prazosin (all at 10(-6) M). Noradrenaline induced concentration-dependent contractions with an EC50, of 1.85 x 10(-6) M. The increases in tension induced by electrical stimulation and noradrenaline were of greater magnitude in arteries denuded of endothelium. In segments with endothelium, N(G)-nitro-L-arginine methyl ester (10(-4) M) or indomethacin (10(-5) M) had no effects on the basal tone, but significantly enhanced the neurogenic and noradrenaline-induced contractions. The potentiation by N(G)-nitro-L-arginine methyl ester of electrical stimulation-induced contractile responses was partially reversed by L arginine (10(-4) M). In the presence of N(G)-nitro-L-arginine methyl ester together with indomethacin the electrical stimulation-induced contractile responses were higher than those obtained when only N(G)-nitro-L-arginine methyl ester or indomethacin was used. N(G)-nitro-L-arginine methyl ester and indomethacin did not influence neurogenic-induced contractile responses of endothelium-denuded arteries. The results suggest that endothelial cells of isolated dog pulmonary arteries depress the contractile response to electrical field stimulation of intramural nerves and that endothelium-derived dilator prostaglandins and nitric oxide may interact to inhibit contractile effects of adrenergic stimulation. PMID- 10382863 TI - Modulation of hypersensitivity reaction by lipids given orally. AB - The effect of lipids administration by gavage (0.4% body weight) given daily during four weeks on the hypersensitivity reaction in trachea, upper and lower bronchi, liver, kidney, mesentery, and pancreas was investigated in male rats. The plasma exudation was assessed by using Evans blue (EB) dye extravasation method. There was a significant difference in the permeability of the organs in nonimmunized rats. The immunization increased the vascular permeability and the response with the organs varied greatly. The effect of lipids on anaphylactic reaction was compared to those of untreated rats (control group). The EB extravasation was significantly increased in the trachea obtained from rats treated with cocoa butter and soybean oil. In the upper bronchi of rats treated with soybean oil, the EB extravasation was increased. However, in the lower bronchi, none of the treatments with lipids changed the extravasation of EB. The same was observed in the liver and kidney. The animals treated with lipids by gavage did not present differences in EB extravasation in the mesentery. However, in the pancreas and duodenum, the treatment with fish and soybean oils and cocoa butter markedly lowered EB extravasation. PMID- 10382862 TI - Preconditioning prevents the negative inotropic action of phenylephrine in rat isolated stunned papillary muscle. AB - The aim of the present study was to establish a model of ischemic preconditioning of rat isolated papillary muscle and to investigate its effect on the simulated ischemia-induced disturbances in contractility and responsiveness to isoproterenol and phenylephrine. Experiments were performed in rat left ventricle papillary muscle. The following parameters were measured: force of contraction (Fc), velocity of contraction (+dF/dt), velocity of relaxation (-dF/dt), time to peak contraction (ttp), and relaxation time at the level of 10% of total amplitude (tt10). After 60 min of simulated ischemia induced by the perfusion of isolated tissues with no-substrate solution aerated by 95% N2/5% CO2, all of the measured parameters were markedly decreased. There was not complete recovery of Fc, +dF/dt and -dF/dt after 60 min of reperfusion. Positive inotropic action of isoproterenol does not differ before and after simulated ischemia. In contrast, phenylephrine induces a positive inotropic action in non-treated, but a significant negative one in simulated ischaemia/reperfusion treated preparations. The latter effect of phenylephrine was reversed by chloroethylclonidine (CEC), a selective blocker of alpha1b-adrenoceptor, but not by WB-4101, a selective blocker of alpha1a-adrenoceptor. Ischemic preconditioning of rat isolated cardiac tissue induced by the 5 min perfusion with no-substrate solution, aerated by 95% N2/ 5% CO2, in the presence of fast electrical pacing (BCL shortened from 2000 ms to 700 ms) and 10 min reperfusion, significantly improves a recovery of the contractility and prevents phenylephrine negative inotropic action. PMID- 10382864 TI - Salviae radix root extract inhibits immunoglobulin E-mediated allergic reaction. AB - We investigated the effects of the aqueous extract of Salviae radix root (SRRAE) on immediate allergic reactions. SRRAE inhibited by 72.7% passive cutaneous anaphylaxis activated by anti-dinitrophenyl (DNP) immunoglobulin E (IgE). SRRAE dose dependently inhibited histamine release and tumor necrosis factor-alpha production from the rat peritoneal mast cells (RPMCs) by anti-DNP IgE. However, SRRAE showed no significant inhibitory effect on compound 48/80-induced systemic allergic reaction and histamine release from RPMCs. The level of cAMP in RPMCs, when SRRAE was added, significantly increased compared with that of a normal control. These results indicate that SRRAE may contain compounds with actions that inhibit anti-DNP IgE-induced mast cell degranulation in rats. PMID- 10382865 TI - Possible interaction between new quinolones and immune functions in macrophages. AB - Some of the immunological effects of a variety of new quinolones on chemotaxis and the production of superoxide anion in rat macrophages were studied. All of the new quinolones examined at a dose range of 0.5 to 50 microg/ml significantly inhibited chemotaxis in a dose-dependent manner in rat macrophages. Moreover, the new quinolones at a dose of 0.5 microg/ml were effective in markedly potentiating the generation of superoxide anion. These results indicate that the new quinolones may modulate immune functions in rat macrophages. PMID- 10382866 TI - Aflatoxin B1 is an inhibitor of cyclic nucleotide phosphodiesterase activity. AB - Aflatoxin B1 (AFB1) action on cyclic nucleotide phosphodiesterase (PDE) activity has been tested on tissue extracts of various organs. In the presence of 100 microM AFB1 a significant inhibition of cAMP and cGMP hydrolytic activity is observed in all tested tissue extracts. However, cGMP hydrolytic activity appears more sensitive to AFB1 inhibition than cAMP hydrolytic activity and a considerably higher inhibition is observed in lung and spleen, than in liver, brain, kidney, and heart. When cGMP is used as substrate, the inhibitory response reaches 72% in lung and spleen extracts. We have also tested AFB1 effects on lung and liver PDE activity peaks separated by DEAE-cellulose chromatography. These data confirm the poor sensitivity to the toxin of all PDE activities present in liver, while the lung peak (where PDE V in present) shows a higher sensitivity to AFB1. In order to establish whether PDE V is in fact more sensitive to AFB1, we have used mouse neuroblastoma cells, in which cGMP hydrolytic activity has been shown to be due to PDE V only. In this case, the calculated IC50 is 24 microM and Dixon plot analysis shows a competitive inhibitory effect with a Ki of 16.7 microM. We have also used aflatoxin B2 and M2, and they proved to be much less effective than AFB1: AFB2 inhibits PDE V with an IC50 of 117 microM, while AFM2 does not show any effect. These results provide the first evidence of a competitive inhibition of AFB1 on an enzymatic activity and suggest that an alteration of cellular cyclic nucleotide levels may play a role in the mechanism of aflatoxin action. PMID- 10382867 TI - Palm vitamin E improves bone metabolism and survival rate in thyrotoxic rats. AB - The effects of vitamin E derived from palm oil on bone turnover in thyrotoxic rats were studied. Palm vitamin E reduced bone resorption to a greater extent than bone formation in thyrotoxic rats, suggesting a net reduction in bone loss. The action of palm vitamin E is probably due to its antioxidant properties. Survival rates were also significantly increased in thyrotoxic rats given palm vitamin E, suggesting the role of free radicals in the overall morbidity and mortality in thyrotoxicosis. PMID- 10382868 TI - Does prevention of free radical reactions influence digoxin-arrhythmias? AB - In the present study, the possible role of oxygen-derived free radicals (OFR) on digoxin- (0.6 mg/kg(-1) IV bolus) induced arrhythmias of anesthetized guinea-pigs has been investigated. Guinea-pigs (300400 g) of either sex were anesthetized with urethane (1.5 g/kg(-1),IP), and their trachea for respiration, left common carotid artery for blood pressure monitoring, and right jugular vein for drug administration were cannulated. ECG and haemodynamics were recorded throughout the experiments. None of the agents used [N-acetyl-L-cysteine (20 mg/kg(-1)IV bolus), or SOD (30,000 IU/kg(-1) IV bolus) + catalase (15,000 IU/kg(-1) IV bolus)] significantly inhibited the arrhythmias except desferrioxamine which reduced the incidence of ventricular fibrillation and arrhythmia score. Desferrioxamine, by acting intracellularly unlike other agents used, might prevent the reduction of Fe(+3) by ascorbate and superoxide anion thus inhibiting the formation of cytotoxic hydroxyl radical in this experimental setting. PMID- 10382869 TI - A review of the comparative in-vitro activities of 12 antimicrobial agents, with a focus on five new respiratory quinolones'. AB - The efficacies of many antimicrobial agents are being threatened by a global increase in the numbers of resistant bacterial pathogens--microorganisms that were once susceptible to some of these agents. In particular, antimicrobial resistance amongst strains of Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pneumoniae has limited the usefulness of first-line agents in some clinical settings. Quinolones were introduced in the 1980s and represented a significant therapeutic advancement in the treatment of patients with infectious diseases. While these compounds possessed potent in-vitro activities against a wide range of gram-negative pathogens, their activities against some gram positive and 'atypical' pathogens remained borderline. Further advancement in the development of quinolones has overcome some of these problems. The 'respiratory quinolones' represent a new generation within this class of molecules and comprise compounds possessing broad spectrum activities against gram-negative, gram-positive and atypical pathogens. This review will focus on the in-vitro activities of five new respiratory quinolones (gatifloxacin, grepafloxacin, levofloxacin, moxifloxacin and trovafloxacin), ciprofloxacin and six non quinolone agents (azithromycin, clarithromycin, amoxycillin, co-amoxiclav, cefuroxime and co-trimoxazole) against a range of bacterial and atypical pathogens, including those that are now resistant to several of these compounds. PMID- 10382870 TI - Comparative susceptibility to penicillin and quinolones of 1385 Streptococcus pneumoniae isolates. Austrian Bacterial Surveillance Network. AB - Antibiotic resistance among Streptococcus pneumoniae isolates has spread rapidly throughout the world since the first description of a strain with diminished susceptibility to penicillin in Australia in 1967. A total of 1385 strains of S. pneumoniae, collected in several centres throughout Austria, were assessed for their sensitivity to moxifloxacin, trovafloxacin, ciprofloxacin, ofloxacin, levofloxacin and lomefloxacin. The MICs were determined using the agar dilution method, according to NCCLS guidelines. Both moxlfloxacin and trovafloxacin showed good anti-pneumococcal activity in terms of MIC50 (both 0.125 mg/L) and MIC90 (both 0.25 mg/L). Less active, but with similar activity to each other, were ciprofloxacin and levofloxacin, each with an MIC50 of 1 mg/L and an MIC90 of 2 mg/L. Ofloxacin showed only moderate activity (MIC50, 1 mg/L; MIC90, 2 mg/L) and lomefloxacin was the least active compound (MIC50, 4 mg/L; MIC90, 8 mg/L). Both moxifloxacin and trovafloxacin at a concentration of < or = 0.5 mg/L inhibited all of the S. pneumoniae strains tested. PMID- 10382871 TI - In-vitro activity of moxifloxacin against fluoroquinolone-resistant strains of aerobic gram-negative bacilli and Enterococcus faecalis. AB - MICs of the new fluoroquinolone, moxifloxacin, and those of ciprofloxacin, ofloxacin and sparfloxacin for 19 genetically characterized fluoroquinolone resistant strains were determined by the agar dilution method. The MICs of moxifloxacin for Escherichia isolates with one mutation in gyrA (corresponding to Ser83-->Leu or Asp87-->Gly substitution) were 0.25-0.5 mg/L, while those of ciprofloxacin, ofloxacin and sparfloxacin were 0.06-0.25, 1 and 0.12-0.5 mg/L, respectively. These values were four- to 16-fold higher than those of the same antibiotics for the wild-type strain, E. coli KL16. Similar results were observed with clinical isolates of Salmonella spp. harbouring one mutation in gyrA leading to the substitution of Ser83 by Phe or Tyr. In the presence of two mutations in the E. coli gyrA gene, the MICs of moxifloxacin ciprofloxacin, ofloxacin and sparfloxacin were 2, 0.5, 4 and 1 mg/L, respectively; these were 32 times higher than the MICs of these agents for E. coli KL16. The MICs of the four quinolones for triple mutants with two mutations in gyrA and one in parC were even higher, i.e. 8, 8, 16 and 8-16 mg/L, respectively. The MICs of moxifloxacin for Campylobacter coli and Campylobacter jejuni strains with a gyrA mutation leading to Thr86-->Ile substitution ranged from 1 to 2 mg/L, while the MICs of ciprofloxacin, ofloxacin and sparfloxacin were 16-32 mg/L, 8-16 and 4-8 mg/L, respectively. For high-level ciprofloxacin-resistant (MICs of 32 mg/L) clinical isolates of Enterococcus faecalis with one substitution at position 83 in GyrA (E. coli coordinates), the MICs of moxifloxacin, ofloxacin and sparfloxacin were 8-16, > or = 128 and 32 mg/L respectively. In conclusion, moxifloxacin and other fluoroquinolones exhibit cross-resistance against aerobic gram-negative bacilli and enterococci. The in-vitro activity of moxifloxacin was greater than that of ofloxacin and slightly less than that of ciprofloxacin and sparfloxacin against Enterobacteriaceae, but greater than those of the three other compounds tested against Campylobacter spp and E. faecalis. PMID- 10382872 TI - The in-vitro activity of moxifloxacin against Legionella species and the effects of medium on susceptibility test results. AB - The in-vitro activities of moxifloxacin, ciprofloxacin, erythromycin and rifampicin against 49 Legionella spp. isolates were determined by an agar dilution method with buffered charcoal yeast extract agar containing alpha ketoglutarate. Because the inhibitory effects of charcoal in the test media were pronounced (92% for quinolones, 90.5% for rifampicin and 92.5% for erythromycin), the MICs were corrected for the charcoal-bound fraction of the antibiotics. The corrected geometric mean MICs were 0.018 mg/L for moxifloxacin, 0.02 mg/L for ciprofloxacin, 0.27 mg/L for erythromycin and 0.005 mg/L for rifampicin. PMID- 10382873 TI - The effect of moxifloxacin on its target topoisomerases from Escherichia coli and Staphylococcus aureus. AB - The effect of moxifloxacin on its target enzymes was evaluated by three different approaches: (i) the MICs of moxifloxacin and nine other fluoroquinolones were determined for mutants of Escherichia coli (n = 13) and Staphylococcus aureus (n = 5) carrying different combinations of resistance mutations; (ii) the activity of moxifloxacin on isolated targets was determined as IC50 values for wild-type and mutant type II topoisomerases from E. coli; and (iii) the mutation frequencies were determined for two single-step mutants (MI with a Ser83-->Leu mutation in gyrA and WT-4 with a Ser80-->Ile mutation in parC) and their parent strain (WT). Of the quinolones tested, moxifloxacin was the only one showing an equivalent high activity against both targets. This is reflected by a comparable high susceptibility of the test strains of E. coli and S. aureus and by the IC50 values of moxifloxacin which were 50-90% lower than those of ciprofloxacin, norfloxacin and sparfloxacin for the wild-type and single mutant enzymes of gyrase and topoisomerase IV. However, double mutant GyrA was significantly more sensitive to moxifloxacin than to the other fluoroquinolones tested, while wild type topoisomerase IV was two-fold more refractory. Mutation rates of WT, MI and WT-4 for ciprofloxacin and moxifloxacin were 5 x 10(-8) vs 4 x 10(-10); <6 x 10( 11) vs <6 x 10(-11); and 2 x 10(-6) vs 5 x 10(-7), respectively. These data indicate an equivalent high inhibitory activity of moxifloxacin on DNA gyrase and topoisomerase IV of E. coli. PMID- 10382874 TI - Antimicrobial activity and accumulation of moxifloxacin in quinolone-susceptible bacteria. AB - The antibacterial activity of moxifloxacin, compared with that of ciprofloxacin, was determined for five strains of Staphylococcus aureus, including one NorA overproducing strain, two quinolone-susceptible strains of Streptococcus pneumoniae, four quinolone-susceptible strains of Haemophilus influenzae, and one strain each of quinolone-susceptible Escherichia coli, Pseudomonas aeruginosa and Moraxella catarrhalis. In addition, the accumulation of moxifloxacin and ciprofloxacin by the NCTC type strain of S. pneumoniae, H. influenzae, S. aureus, E. coli and P. aeruginosa was determined by a fluorescence method. For all strains, moxifloxacin accumulated to a lower concentration than ciprofloxacin. The concentrations of moxifloxacin accumulated ranged from 12 to 44 ng/mg dry cells. The lowest concentration was accumulated by S. pneumoniae NCTC 7465 and the highest concentration by S. aureus NCTC 8532. Increased expression of norA in S. aureus had no effect on the accumulation of moxifloxacin. Despite differences in the concentration of moxifloxacin accumulated by the different species, there was little difference between the MICs of this agent for each strain (0.06-0.5 mg/L), suggesting that the concentration accumulated by wild-type bacteria has little effect on the MIC. PMID- 10382875 TI - Bactericidal properties of moxifloxacin and post-antibiotic effect. AB - The time-kill kinetics and post-antibiotic effect (PAE) of moxifloxacin were studied for strains of Streptococcus pneumoniae, Streptococcus pyogenes, Haemophilus influenzae, Staphylococcus aureus and Escherichia coli. Moxifloxacin had a bactericidal effect against all strains tested, with the least rapid bactericidal effect being against S. pyogenes and the most rapid effect against S. aureus and E. coli. The PAE of moxifloxacin was similar to that of other fluoroquinolones and increased with increasing concentration. No association was found between the bactericidal effect of moxifloxacin and the duration of PAE. Gram-positive and gram-negative organisms were also exposed to concentrations of moxifloxacin, sparfloxacin and amoxycillin that simulated the drug concentrations obtained in human serum after standard oral dosing schedules. Simulation of moxifloxacin concentrations in human serum reduced viable counts more effectively and more rapidly than shown in time-kill experiments; in contrast, sparfloxacin and amoxycillin were less effective than when constant concentrations of these antibacterials were used. PMID- 10382876 TI - Pharmacodynamics of fluoroquinolones. AB - Fluctuating concentrations of three fluoroquinolones (moxifloxacin, sparfloxacin and ofloxacin) and a beta-lactam (amoxycillin) were used in vitro to simulate antibiotic concentrations in human serum after oral doses of antibiotics. The antibiotics were tested against Staphylococcus aureus 12241 and Streptococcus pneumoniae 4241. Moxifloxacin and sparfloxacin were also tested against Escherichia coli Neumann. Human serum concentrations of moxifloxacin and ciprofloxacin were also simulated in an in-vivo murine thigh muscle model against S. aureus, S. pneumoniae and E. coli. Ciprofloxacin, sparfloxacin and ofloxacin had a dose-independent effect on gram-positive organisms beyond their optimal dose that gave a maximum effect, as did amoxycillin. In contrast, moxifloxacin had a dose-dependent and therefore concentration-dependent effect on both gram positive and beta-lactam-susceptible and-resistant gram-negative organisms. The marked activity of moxifloxacin against both gram-positive and gram-negative organisms was confirmed in an in-vivo model. A human dose equivalent of 200 mg moxifloxacin reduced viable counts of S. pneumoniae below the limit of detection and regrowth did not occur. S. aureus was eliminated almost as effectively as S. pneumoniae. A 200 mg dose of moxifloxacin completely eliminated the original inoculum of E. coli within 6 h. Treatment of S. aureus with ciprofloxacin (250 or 500 mg) resulted in a dose-independent decrease in viable counts by approximately 3.5 log10 cfu/mL. A 125 mg dose of ciprofloxacin almost completely eliminated the original inoculum of E. coli within 8 h, whereas both the 250 mg and 500 mg doses reduced viable counts below the limit of detection. Thus, the in-vitro and in vivo pharmacodynamic models used in this study established that moxifloxacin was highly effective against both gram-negative and gram-positive bacteria. PMID- 10382877 TI - Pharmacokinetics of the 8-methoxyquinolone, moxifloxacin: tissue distribution in male rats. AB - BAY 12-8039 (moxifloxacin-HCl) and 14C-labelled BAY 12-8039 were administered to male rats as single i.v. and oral doses of 4.6 and 5.0 mg/kg bodyweight respectively. The distribution of substance-associated radioactivity in the body was investigated by whole-body autoradiography. The concentrations of the unchanged compound in plasma, skin suction blister fluid and lung tissue were determined by HPLC. Whole-body autoradiography revealed distinctly higher concentrations of radioactivity in the gastrointestinal tract, urinary bladder and in most organs and tissues (e.g. kidneys, liver, spleen, lungs, various glands, cartilaginous tissues and in melanin-containing structures located in the eye, meninges and hair follicles of pigmented skin) than in blood. Radioactivity crossed the blood-brain barrier only to a small extent. The results show a high tissue affinity and a rapid and homogeneous distribution of radioactivity from blood to organs or tissues. No relevant difference in the distribution of radioactivity was found following i.v. and oral administration. After i.v. and oral dosing similar concentrations of the unchanged compound were determined in skin suction blister fluid and plasma. The concentrations of the unchanged compound in lung tissue were about three times higher than those in plasma following both i.v. and oral administration. The concentration-time courses for moxifloxacin in plasma and lung tissue were parallel. PMID- 10382879 TI - Fluoroquinolone phototoxicity: a comparison of moxifloxacin and lomefloxacin in normal volunteers. AB - Moxifloxacin, a broad-spectrum fluoroquinolone with the methoxy group at position 8 of the quinolone structure that is believed to confer reduced phototoxicity, was investigated in 32 healthy human male volunteers by a randomized double-blind placebo and positive control (lomefloxacin) phototest technique. A comparison of pre- and on-drug photosensitivity levels tested with an irradiation monochromator using relevant sunlight wavelengths, failed to demonstrate phototoxicity after administration of either placebo or moxifloxacin (200 mg or 400 mg/day) for 7 days. As expected, lomefloxacin (400 mg/day) phototoxicity was revealed at the UVA wavebands 335 +/- 30 nm and 365 +/- 30 nm (maximal at 24 h), with a phototoxic index of 3-4. The susceptibility to this effect rapidly normalized within 48 h of stopping the drug. No special protection from UVA wavelengths is necessary for those taking moxifloxacin. PMID- 10382878 TI - Pharmacokinetics of the 8-methoxyquinolone, moxifloxacin: a comparison in humans and other mammalian species. AB - The pharmacokinetics of moxifloxacin was investigated in NMRI mice, Wistar rats, rhesus monkeys, beagle dogs, Gottingen minipigs and healthy human volunteers after i.v. and oral administration of moxifloxacin-HCl (single doses of moxifloxacin 9.2 mg/kg bodyweight) in animals and 100 mg moxifloxacin (1.4 mg/kg bodyweight p.o. and 1.2 mg/kg bodyweight i.v.) in humans. The plasma concentration vs time courses of the unchanged compound (determined by HPLC) and the derived pharmacokinetic parameters were used to evaluate the absorption process, to compare the pharmacokinetics in these species and to perform an interspecies scaling. The results of the pharmacokinetic investigations indicate a clear dependence on the species. Moxifloxacin is absorbed quickly (rats, dogs, humans > monkeys): the major portion of the dose reached the systemic circulation within the first 2 h. In the minipig absorption was slower. Bioavailability was high to moderate (91-52%) in all species. Protein binding (f(u)) was low (55-71%) in all species. The volume of distribution at steady state (Vss) was medium to large (2.0-4.9 L/kg) in all species. There were considerable differences in maximum concentrations (C(max,norm), 0.430-0.070 kg/L) and in AUCnorm values (oral, 6.18-0.184 kg x h/L; i.v., 7.51-0.237 kg x h/L). Total body clearance (CL) decreased with increasing bodyweight (4.21-0.132 L/(h x kg)). The mean residence time (MRT) decreased with decreasing bodyweight (15-0.88 h). The half-life (t(1/2)) decreased with decreasing bodyweight (oral, 12-1.3 h, i.v., 13-0.93 h). There was moderate to low renal excretion (i.v., 20-6.2%), the renal clearance, (CL(R)) was in the range 0.615-0.0222 L/(h x kg). Regarding the pharmacokinetic parameters determined after oral administration, the dog was most similar to the human in terms of Cmax, AUC and t(1/2). There was good correlation between bodyweight and CL (coefficient of correlation (r) = 0.959), Vss (r = 0.990) and MRT (r = 0.943). On the basis of preclinical studies a terminal half-life appropriate for once-daily dosing in humans was predicted and confirmed by Phase I data. PMID- 10382880 TI - Pharmacokinetics and elimination of moxifloxacin after oral and intravenous administration in man. AB - The pharmacokinetics of moxifloxacin and its metabolites M1 (sulpho-compound) and M2 (acyl-glucuronide) were characterized in 12 healthy male volunteers in an open, randomized, crossover study. After an overnight fast the volunteers were given a single 400 mg dosage of moxifloxacin either as a tablet or a 1 h infusion with a washout phase of at least 1 week between the two treatments. Multiple plasma, faeces and urine samples were collected for the analysis of moxifloxacin and metabolites using validated HPLC with fluorescence detection. The AUC for both formulations was comparable with bioequivalence criteria fulfilled, with Cmax after oral treatment approximately 31% lower. Following oral administration, absorption was fast with low to medium variability (mean dissolution and absorption time 2.4 h). The absolute bioavailability was 86%. The excretion of moxifloxacin and its metabolites was quantified in a subset of eight subjects. More than 96% of the dose was recovered from urine and faeces after oral dosing, and >98% following i.v. administration of the drug. M1, which is strongly bound to plasma proteins (90%), was mainly eliminated into faeces (approximately 37-38% of the administered dose) and to a minor extent into urine (2.5% of the administered dose) by active tubular secretion. M2 (only 5% bound to plasma protein) was only found in urine, where it amounted to approximately 14% of the dose. Plasma concentrations of the metabolites were much lower than those of the parent compound. Moxifloxacin was well tolerated with few adverse events and no clinically relevant changes in laboratory values. PMID- 10382881 TI - Preclinical safety evaluation of moxifloxacin, a novel fluoroquinolone. AB - The toxicity of moxifloxacin (BAY 12-8039), a novel fluoroquinolone with a broad spectrum of antibacterial activity, was evaluated in a comprehensive programme of toxicological studies that included single and multiple dose toxicity studies in rats, mice, dogs and monkeys, reproductive system toxicity studies in rats and monkeys and in-vitro and in-vivo mutagenicity assays. Although moxifloxacin is not intended for long-term clinical use, an accelerated bioassay in target organs to assess carcinogenesis was performed in rats. In addition to the routine toxicological programme required for the development of a drug intended for short term administration, a major part of the preclinical programme for moxifloxacin comprised studies designed specifically to address the safety issues known to be features of fluoroquinolones, i.e. adverse effects on the central nervous and cardiovascular systems, phototoxicity, arthrotoxicity and oculotoxicity. The results of the toxicological investigations confirmed that the safety profile of moxifloxacin is comparable to those of other fluoroquinolones. PMID- 10382882 TI - Antimycobacterial activities of riminophenazines. AB - Riminophenazines were specifically developed as drugs active against Mycobacterium tuberculosis but extensive research over several decades has shown that these compounds are also active against many other mycobacterial infections, particularly those caused by Mycobacterium leprae and the Mycobacterium avium complex (MAC). Clofazimine, the lead compound in this series, is included in the regimens that are approved by the WHO for the treatment of leprosy and has contributed significantly to the control of that disease, particularly that caused by dapsone-resistant bacteria. Despite early problems, clofazimine has shown clinical efficacy in tuberculosis, in particular that caused by multiple drug resistant strains. Clofazimine does not induce resistance and also inhibits emergence of resistance to isoniazid in M. tuberculosis. The efficacy of clofazimine against MAC is more varied and the availability of better drugs has limited its use. Newer riminophenazines, such as B746 and B4157, not only showed increased anti-mycobacterial activity but also produced less skin pigmentation, which is the main drawback of this group of compounds. The most important virtues of riminophenazines, such as intracellular accumulation in mononuclear phagocytic cells, anti-inflammatory activity, a low incidence of drug resistance and slow metabolic elimination, make them attractive candidates for the treatment of mycobacterial infections. It is essential, however, to investigate the newer analogues clinically, while continuing the pursuit of alternate candidates that demonstrate higher anti-mycobacterial activity and lower rates of skin pigmentation. PMID- 10382883 TI - Plasmid heterogeneity and identification of a Tn5281-like element in clinical isolates of high-level gentamicin-resistant Enterococcus faecium isolated in the UK. AB - Ten clinical isolates of high-level gentamicin-resistant (HLGR) Enterococcus faecium, collected from six hospitals throughout the UK, were studied to determine whether HLGR was attributed to widespread dissemination of a single plasmid or whether different plasmid types were implicated in the dissemination of this phenotype. HLGR was attributed to the presence of the AAC6'-APH2" bifunctional aminoglycoside modifying enzyme. The aac6'-aph2" gene was present on a 70 kb plasmid in all ten isolates. Conjugation studies indicated that the HLGR marker could transfer with varying frequency, with or without the associated 70 kb plasmid. Detailed molecular genetic analysis suggested that four of the isolates harboured a transposon similar to Tn5281, originally identified in Enterococcus faecalis strain HH22 isolated in the USA. The UK transposon, however, lacked the two symmetrically located HaeIII sites found in Tn5281. The six remaining isolates appeared to have a Tn5281-truncated structure in which the aac6'-aph2" gene is flanked by an IS256 element at the 5' end. Further studies with nine restriction endonucleases showed that the aac6'-aph2" gene was associated with two different plasmid types in E. faecium. Pulsed-field gel electrophoresis (PFGE) analysis identified three different patterns. The four E. faecium isolates harbouring the Tn5281-like structure were indistinguishable from each other, while the remaining six isolates exhibited two distinct PFGE patterns. This is the first study to indicate that there is heterogeneity among the plasmids that confer the HLGR phenotype in E. faecium isolates in the UK and to report on the presence of a transposon, similar to Tn5281, in E. faecium harbouring the aac6'-aph2" gene. PMID- 10382884 TI - Distribution of mefE and ermB genes in macrolide-resistant strains of Streptococcus pneumoniae and their variable susceptibility to various antibiotics. AB - From February to October 1995, 62 erythromycin-resistant strains of Streptococcus pneumoniae isolated at Yamanashi Red Cross Hospital were tested to determine their susceptibility to various macrolides, subjected to resistance induction tests by the disc diffusion method and analysed for genes encoding resistance to macrolides (ermB and mefE). On the basis of resistance induction testing, the isolates were classified as having either inducible (59.7%) or non-inducible (40.3%) macrolide resistance. The ermB gene was always detected in resistance inducible type isolates, either alone or in combination with mefE. The mefE gene alone was found only in non-inducible type isolates. Isolates with non-inducible resistance (those with only the mefE gene) had an intermediate level of resistance to 14-membered macrolides, and were susceptible to rokitamycin, a 16 membered macrolide. According to NCCLS guidelines, 9.6% of S. pneumoniae strains were judged to be susceptible to penicillin, 62.9% of reduced susceptibility and 27.4% penicillin resistant. No correlation was detected between the presence of particular macrolide-resistance genes (ermB, ermB + mefE, or mefE) and resistance to penicillin G. PMID- 10382885 TI - Anti-pneumococcal activity of gatifloxacin compared with other quinolone and non quinolone agents. AB - An agar dilution MIC method was used to test the activity of gatifloxacin, a new broad-spectrum fluoroquinolone, compared with ciprofloxacin, levofloxacin, sparfloxacin, trovafloxacin, amoxycillin, cefuroxime, ceftriaxone and clarithromycin against 71 penicillin-susceptible, 81 penicillin-intermediate and 55 penicillin-resistant pneumococci. Quinolone activity was unaffected by penicillin susceptibility, with MIC50/MIC90s (mg/L) of 0.25/0.5 for gatifloxacin; 1/2 for ciprofloxacin; 1/2 for levofloxacin; 0.25/0.5 for sparfloxacin; 0.125/0.25 for trovafloxacin. beta-Lactam and clarithromycin MICs rose with those of penicillin G; MIC50/MIC90 values (mg/L) for penicillin-susceptible, intermediate and -resistant strains were: 0.03/0.06, 0.25/1, 2/4 for penicillin G; 0.03/0.03, 0.125/1, 2/4 for amoxycillin; 0.03/0.125, 0.5/4, 8/16 for cefuroxime; 0.03/0.03, 0.25/0.5, 2/4 for ceftriaxone; 0.03/0.06, 0.03/>64, 1/>64 for clarithromycin. Time-kill testing of four penicillin-susceptible, four intermediate and four -resistant strains showed that levofloxacin at the MIC, gatifloxacin and sparfloxacin at 2 x MIC, and trovafloxacin and ciprofloxacin at 4 x MIC, were bactericidal (99.9% killing) for all strains after 12 h and 24 h. By contrast, amoxycillin, cefuroxime and ceftriaxone showed bactericidal activity after 24 h against all strains at 4, 8 and 4 x MIC, respectively. Against ten organisms with clarithromycin MICs of 0.03-4.0 mg/L, clarithromycin was bactericidal against seven strains at 8 x MIC after 24 h. Quinolones showed more rapid killing at lower concentrations and earlier time periods than did beta lactams and clarithromycin. PMID- 10382886 TI - Investigation of the synergic effects of aminoglycoside-fluoroquinolone and third generation cephalosporin combinations against clinical isolates of Pseudomonas spp. AB - Antimicrobial synergy resulting from antibiotic combination therapy is often important in the treatment of serious bacterial infections. Previous studies have demonstrated synergy between an aminoglycoside and beta-lactam antibiotics in the treatment of Pseudomonas aeruginosa infections. The present paper investigates the synergic effects of aminoglycosides (amikacin and netilmicin) and fluoroquinolones (ciprofloxacin, ofloxacin and pefloxacin) in combination with third-generation cephalosporins (cefoperazone, ceftriaxone and ceftazidime) against 18 clinical isolates of Pseudomonas spp. The effects of these drugs were examined by three methods (disc diffusion, 'chequerboard' titration and the time killing method), to evaluate the activities of the antibiotics alone and in combination against selected isolates. Fractional inhibitory concentration indices were calculated for all isolates with all combinations. Use of the disc diffusion method revealed that amikacin and netilmicin in combination with the three cephalosporins exhibited synergy against 7-12 isolates, whereas the combinations of quinolones and ceftazidime displayed synergic effects only in the case of 3-5 isolates. On 'chequerboard' titration, amikacin and ceftriaxone exerted synergy against seven of the isolates. The other combinations showed synergy against fewer isolates, but every combination demonstrated synergic effect against some of the isolates. The tested combinations had different effects against various Pseudomonas spp. With the time-killing method, the 1/2 x MIC of amikacin or ciprofloxacin in combination with the 1/2 x MIC of third generation cephalosporins proved to be most effective. No antagonism was found with these combinations. Discrepancies in the detection of synergy were observed for the different methods. PMID- 10382887 TI - Bismuth-mediated disruption of the glycocalyx-cell wall of Helicobacter pylori: ultrastructural evidence for a mechanism of action for bismuth salts. AB - The mechanism of bismuth's bactericidal activity against Helicobacter pylori was investigated using transmission electron microscopy (TEM) and analytical electron microscopy (AEM); time-kill kinetic methods evaluated the effect of excess divalent cations. TEM analysis of untreated H. pylori revealed a normal morphology. In contrast, H. pylori exposed to bismuth salts had swollen, distorted cells with membrane-cell wall blebbing and a cytoplasm containing electron-dense, sometimes crystalline aggregates. By AEM, swollen cells contained bismuth at the cell periphery, whereas bacillary forms contained cytoplasmic bismuth localizations. Time-kill studies showed that the bactericidal activity of bismuth could be prevented by pretreatment with divalent cations. The effects of bismuth salts on the glycocalyces-cell walls of H. pylori with reversal of bactericidal activity by divalent cations are identical to those produced by other polycationic agents on various gram-negative bacilli. We conclude that disruption of the glycocalyces-cell walls of H. pylori is one mechanism of action for bismuth salts. PMID- 10382888 TI - Differential inhibitory effects of protoberberines on sterol and chitin biosyntheses in Candida albicans. AB - The anti-Candida potentials of 12 Korean medicinal plants were explored: methanol extracts from Coptis rhizoma and Phellodendron amurense caused significant inhibition of growth of Candida albicans, Candida glabrata, Candida krusei and Candida parapsilosis. The predominant active components of the extracts were the protoberberines berberine and palmatine; the most potent inhibition of growth was exhibited by berberine on C. krusei (MIC <4 mg/L) and palmatine on C. parapsilosis (MIC 16 mg/L). Both berberine and palmatine inhibited the in-vivo rate of incorporation of L-[methyl-14C]methionine into C-24 of ergosterol in C. albicans (50% inhibition concentration (IC50 values), 25 microM and 300 microM, respectively); this result suggests that sterol 24-methyl transferase (24-SMT) is one of the cellular targets for the antifungal activity of the protoberberines. In-vitro 24-SMT activity in microsomes from the yeast growth form of C. albicans was inhibited by both berberine (inhibition constant (Ki) 232 microM) and palmatine (Ki 257 microM) in a non-competitive manner; inhibition of 24-SMT was more marked for the mycelial form than for the yeast growth form of this organism. Palmatine inhibited chitin synthase from both the yeast and mycelial growth phases of C. albicans in a non-competitive manner (Ki 780 microM). The effects of protoberberines, extracted from established medicinal plants, on both sterol and cell wall biosyntheses in pathogenic fungi indicate that the potential of these compounds, or their semi-synthetic derivatives, as a novel class of antifungal agents should be investigated more fully. PMID- 10382889 TI - Comparison of the toxicity of fluconazole and other azole antifungal drugs to murine and human granulocyte-macrophage progenitor cells in vitro. AB - We studied the inhibitory effects on colony formation by granulocyte-macrophage colony forming units (cfu-gm) of eight azole antifungal agents in vitro. All agents, except fluconazole, inhibited colony formation dose-dependently with 50% inhibitory concentrations (IC50) in the range of 0.78-49 micromol/L in cultures of murine and human bone marrow. For human cells, the IC50 values were 0.553 mg/L for itraconazole, 1.24 mg/L for saperconazole, 2.58 mg/L for clotrimazole, 5.33 mg/L for miconazole, 6.17 mg/L for econazole, 6.27 mg/L for ketoconazole and 8.38 mg/L for oxiconazole. The IC50 of itraconazole for human cfu-gm in vitro was similar to the plasma level of this drug recommended for systemic antifungal therapy (>0.5 mg/L) thus indicating the potential clinical relevance of our data. The IC50 of ketoconazole for human cfu-gm in vitro may be exceeded by plasma levels produced in vivo by high (> or =400 mg) doses, whereas fluconazole failed to reduce colony formation by 50% even at 100 mg/L, a concentration not reached in vivo even after extremely high doses (2000 mg/day). To most of the drugs studied, murine progenitor cells seemed to be less sensitive than the human ones. There was, however, a close correlation between the murine and human log IC50 values of the drugs (r2 = 0.964, P< 0.001), suggesting that cultures of murine bone marrow may be suitable to predict the in-vitro toxicity of azole antifungals to human cfu-gm. PMID- 10382890 TI - Pharmacodynamics of trovafloxacin in experimental pneumococcal meningitis: basis for dosage selection in children with meningitis. AB - Trovafloxacin is a recently approved fluoroquinolone with excellent activity against gram-positive and gram-negative organisms that offers a potential alternative for treatment of beta-lactam-resistant pneumococcal meningitis. Using the rabbit meningitis model, we sought to characterize the pharmacodynamic properties of trovafloxacin in the cerebrospinal fluid (CSF). Animals were given single doses of trovafloxacin of 10, 15, 20 or 30 mg/kg; 1 h after Infusion mean CSF concentrations were 0.59+/-0.18, 0.74+/-0.14, 1.12+/-0.12 and 1.07+/-0.35 mg/L, respectively. The bacterial killing rate Increased with increasing dosages of trovafloxacin, indicating that its activity is concentration dependent. All three pharmacodynamic Indices (area under the concentration curve (AUC)/MBC, peak concentration (Cmax)/MBC, and time above MBC (T > MBC)) correlated with bacterial killing; however, AUC/MBC correlated best (r = 0.71). In a second experiment we found comparable bacterial killing with multiple doses of trovafloxacin given either every serum half-life or every two serum half-lives. In both experiments bacterial regrowth occurred when the concentration of trovafloxacin in CSF fell below the MBC. These data have been used in formulating an appropriate regimen for trovafloxacin treatment of bacterial meningitis in children. PMID- 10382891 TI - Antimicrobial resistance in gram-positive pathogens isolated in the UK between October 1996 and January 1997. AB - Antimicrobial resistance in gram-positive pathogens from 30 centres in the UK (ten Teaching, ten Associate Teaching and ten District General Hospitals) was studied over a 4 month period between October 1996 and January 1997. High-level resistance (HLR) and low-level resistance (LLR) to penicillin amongst pneumococci was 3.3% and 3.4%, respectively. However, considerable variation in resistance rates was observed depending on geographical location (LLR range 0-15.4% and HLR range 0-30.8%). Considerable variation in resistance rates was also observed for Staphylococcus aureus to methicillin, with rates ranging from 0% to 56.7% depending on locality. Using conventional MIC methodology, none of the isolates of S. aureus was considered as having reduced sensitivity to vancomycin. However, eight isolates grew on Brain Heart Infusion Agar containing vancomycin (4 mg/L) after prolonged incubation and are therefore worthy of further investigation by electron microscopy. With Enterococcus faecalis, resistance rates were similar between the three types of hospital and only four isolates were considered resistant to glycopeptide antibiotics (one vanA and three vanB phenotype). PMID- 10382892 TI - Mechanisms of resistance to ampicillin, chloramphenicol and quinolones in multiresistant Salmonella typhimurium strains isolated from fish. AB - Mechanisms of antibiotic resistance and epidemiological relationships were investigated for five multiresistant strains of Salmonella typhimurium isolated from fish in India. Four strains showed resistance to nalidixic acid, chloramphenicol, tetracycline, co-trimoxazole, gentamicin and beta-lactam antibiotics. The remaining strain was susceptible to all beta-lactam antibiotics tested and to co-trimoxazole but resistant to the other antibiotics tested. Epidemiological analysis performed by REP-PCR showed that the five isolates belonged to the same clone. Resistance to nalidixic acid was related to a single mutation in the gyrA gene. Chloramphenicol resistance was related to the production of chloramphenicol acetyl-transferase. An OXA-1 beta-lactamase, located in an integron, was responsible for resistance to ampicillin. These results indicate the health hazard posed by the fact that S. typhimurium may acquire or develop several mechanisms of resistance to a variety of antibiotics, including quinolones, and can thus cause disease in humans which may be difficult to treat. PMID- 10382894 TI - Comparison of agar dilution, microdilution, Etest and disc diffusion to test the activity of trovafloxacin against Pseudomonas aeruginosa, methicillin-resistant Staphylococcus aureus and Streptococcus pneumoniae. AB - To investigate the ability of four in-vitro methods to test trovafloxacin activity, this study evaluated susceptibility of 101 isolates of each of Pseudomonas aeruginosa, methicillin-resistant Staphylococcus aureus and Streptococcus pneumoniae to trovafloxacin by agar dilution, microdilution, Etest and disc diffusion methodologies. MIC50 and MIC90 values were very similar for all three species with all four methods. For S. aureus and P. aeruginosa, good correlation was obtained between breakpoints of > or =17 mm, 14-16 mm and < or =13 mm with agar and microdilution MICs. For both species, Etests yielded susceptibility rates lower than the other three methods. For pneumococci, excellent correlation was obtained with all four methods. PMID- 10382893 TI - In-vitro antimicrobial activity of a carbapenem, MK-0826 (L-749,345) and provisional interpretive criteria for disc tests. AB - The in-vitro activity of MK-0826, a new oral carbapenem, was compared with that of imipenem by broth microdilution susceptibility tests against 545 bacterial isolates. MK-0826 had significantly greater activity against Enterobacteriaceae and poorer activity against Pseudomonas aeruginosa and many gram-positive species. MK-0826 disc diffusion tests were also performed according to the NCCLS procedure and tentative interpretive criteria were determined for possible susceptible MIC breakpoints of < or = 4.0 and < or = 2.0 mg/L. PMID- 10382895 TI - In-vitro activity of grepafloxacin, a new fluoroquinolone, against mycoplasmas. AB - The in-vitro activity of grepafloxacin, a new oral fluoroquinolone antibiotic, was compared with those of three other fluoroquinolones and two unrelated antimicrobials, doxycycline and erythromycin, against various Mycoplasma spp. For 65 mycoplasma and 42 ureaplasma strains, grepafloxacin (MIC range 0.03-2 mg/L) was some two to 16 times more active than ofloxacin and ciprofloxacin, showing similar activity to that of sparfloxacin. MBCs of grepafloxacin increased two- to 16-fold when compared with MICs and were comparable to those of sparfloxacin, and lower than those of ofloxacin and ciprofloxacin. PMID- 10382896 TI - Co-amoxiclav affects cytokine production by human polymorphonuclear cells. AB - Some antimicrobial agents have been reported to modify the host immune responses both in vivo and in vitro. As we demonstrated previously that co-amoxiclav had beneficial properties which result in enhancement of the microbicidal functions of human poly-morphonuclear cell (PMNs), we investigated the modulatory effect of this combination on cytokine production by human PMNs in vitro. The addition of co-amoxiclav elicited the production by lipopolysaccharide (LPS)-stimulated PMNs of substantial amounts of some cytokines, namely IL-8 and IL-1beta, after the addition of Klebsiella pneumoniae. These cytokine levels were higher than those obtained by PMNs incubated in culture medium only, without co-amoxiclav. PMID- 10382897 TI - Difficulties in the assay of liposomal amikacin (MiKasome) in serum. AB - Antibiotic-free human serum was spiked with known concentrations of liposomal amikacin and assayed on the Abbott TDx System, using polarization fluoroimmuno assay (PFIA) kits from Abbott Laboratories, Oxis and Sigma. Although all three kits gave a linear response, the Abbott and Oxis kits showed very low recovery (<21%) with only the Sigma kit giving near 100% recovery. Heating samples at 56 degrees C for 30 min improved recovery with the Abbott and Oxis kits (75-80% of target value), but decreased recovery with the Sigma kit (85% of target value). The loss of amikacin from liposomal amikacin, as measured using the Sigma kit, was related to both temperature and duration of heating, reaching a maximal loss of 21% after 1 h at 60 degrees C. PMID- 10382898 TI - Bioassays for itraconazole blood levels: an interlaboratory collaborative study. AB - Duplicate bioassays for itraconazole and hydroxy-itraconazole were run with 30 serum samples in five laboratories, each using a different method. Both itraconazole and hydroxy-itraconazole were used as standards. Despite quantitative variations, the results of the bioassays correlated sufficiently to indicate the relative level of antifungal activity in the test samples. PMID- 10382899 TI - Reduced susceptibility to vancomycin of nosocomial isolates of methicillin resistant Staphylococcus aureus. AB - The MICs of vancomycin for 56 random nosocomial Staphylococcus aureus isolates homogeneously resistant to methicillin (homMRSA), 16 heterogeneously resistant isolates (hetMRSA) and 25 susceptible isolates (MSSA) were determined by a standard broth microdilution method. Representative isolates were also tested by an agar incorporation method, the Etest and population analysis. Although always in the susceptible range, MICs of vancomycin for homMRSA were significantly higher than those for hetMRSA or MSSA. Moreover, a homMRSA strain belonging to one of the major Greek MRSA clones contained a sub-population of cells that could grow in the presence of vancomycin 8 mg/L at a frequency of 6.7 x 10(-8). PMID- 10382901 TI - Efficacy and pharmacodynamics of teicoplanin given daily during the first 3 days and then on alternate days for methicillin-resistant Staphylococcus aureus infections. AB - Fifteen evaluable patients (mean age, 67 years) were enrolled to assess the efficacy of teicoplanin, 6 mg/kg given daily during the first 3 days and then on alternate days, for the treatment of methicillin-resistant Staphylococcus aureus (MRSA) infections. Eight patients had soft tissue infections, four catheter associated bacteraemia, two osteomyelitis and one pneumonia. Clinical cure was observed in 13 of 15 patients. Both clinical and bacteriological failures were shown in the two patients with osteomyelitis. The mean serum levels of teicoplanin (mg/L) were 22, 8 and 6.7 for peak, 24 h and 48 h troughs, respectively. The dosage employed in this study proved effective in non-deep seated MRSA infections. PMID- 10382902 TI - Prevention of febrile leucopenia after chemotherapy in high-risk breast cancer patients: no significant difference between granulocyte-colony stimulating growth factor or ciprofloxacin plus amphotericin B. AB - In a prospective randomized trial, 40 stage IV breast cancer patients undergoing intermediate high-dose chemotherapy (cyclophosphamide, 5-fluorouracil plus epirubicin or methotrexate), received either recombinant human G-CSF (rhG-CSF, group I) or ciprofloxacin and amphotericin B (CAB, group II) for prevention of febrile leucopenia (FL). In group I, seven of 18 patients developed FL (after 10/108 courses); in group II, seven of 22 patients (7/98 courses) (P = NS). Median hospitalization duration and costs were not different. RhG-CSF was 6.6 times more expensive per course than CAB. In conclusion, prophylactic CAB has similar efficacy to rhG-CSF in this setting, and is more cost-effective. PMID- 10382900 TI - Lack of effect of dirithromycin on theophylline pharmacokinetics in healthy volunteers. AB - Twelve healthy volunteers were enrolled in an open-label, randomized, crossover study. Subjects received single doses of theophylline (5 mg/kg) alone and after a 10 day course of dirithromycin (two 250 mg tablets od). The study phases were separated by a 3 week washout period. Serum samples were collected before and for 24 h after theophylline doses. Serum theophylline concentrations were measured via a validated immunoassay system and the data were modelled via non compartmental analysis. When the control phase (i.e. no dirithromycin) was compared with the treatment phase (i.e. with dirithromycin), theophylline exposures as measured by AUC0-->infinity were not significantly different: 141.7+/-25.9 and 136.4+/-33.1 mg x h/L respectively (P = 0.16). No significant changes in other theophylline pharmacokinetic parameters were evident. These results indicate that theophylline can be safely co-administered with dirithromycin. PMID- 10382903 TI - Erythromycin resistance amongst group A beta-haemolytic streptococci isolated in a paediatric hospital in Athens, Greece. PMID- 10382904 TI - Familial infantile myasthenia: confusion in terminology. PMID- 10382905 TI - Congenital myasthenic syndrome caused by a mutation in the Ets-binding site of the promoter region of the acetylcholine receptor epsilon subunit gene. AB - Forty-two missense, truncation, or splice-site mutations of the acetylcholine receptor (AChR) subunit genes have been reported to date in patients with congenital myasthenic syndromes. Here we report a homozygous mutation, epsilon 155G --> A, in the promoter region of the AChR epsilon subunit gene that converts the Ets-binding site of the promoter region from CGGAA to CAGAA. The asymptomatic parents and brother are heterozygous and an affected sister is homozygous for epislon-155G --> A. The Ets-binding site mediates synapse specific expression of the AChR epsilon subunit gene. An identical G-to-A mutation in the mouse Ets binding site was previously shown to decrease the binding affinity of the Ets binding site for the GA binding protein, a transactivating factor for the Ets binding site, and to reduce the synapse specific expression of the epsilon subunit. The decreased synaptic expression of the epsilon subunit readily accounts for the congenital myasthenic phenotype. PMID- 10382906 TI - Adult glycogenosis II with paracrystalline mitochondrial inclusions and Hirano bodies in skeletal muscle. AB - Hirano bodies constitute eosinophilic intracytoplasmic inclusions, typically seen in the central nervous system, where they are related to senility and certain dementias such as Alzheimer's disease or the Parkinson-dementia complex. They have been found in different tissues of experimental animals and, on rare occasions, in extraocular muscles of elderly individuals. However, to our knowledge they have not been described in skeletal muscle in locations other than extraocular muscles or associated with muscle pathology. Glycogenosis II or Pompe's disease, is a metabolic disorder caused by acid maltase deficiency and is characterized by glycogen accumulation in lysosomes in various tissues, including skeletal muscle. There are three clinical forms depending on age at onset, the most frequent being the childhood form. We present the histopathological and ultrastructural findings of a muscle biopsy performed in a case of the adult form of glycogenosis II which showed, in addition to characteristic lysosomal glycogen storage, paracrystalline mitochondrial inclusions and, as an exceptional finding, intracytoplasmic Hirano bodies in some muscle fibres. PMID- 10382907 TI - Proximal myotonic myopathy (PROMM) presenting as myotonia during pregnancy. AB - Proximal myotonic myopathy is a recently described autosomal dominant condition characterized by proximal myopathy, cataracts, intermittent myotonia, and myalgia. We report a further family with this condition. The proband and her two sisters presented with myotonia during pregnancy which resolved after each delivery. Two sisters experienced myalgia between each pregnancy. This relationship between pregnancy and symptom exacerbation suggests an intriguing hormonal influence in PROMM. PMID- 10382908 TI - Correlation of muscle fiber type measurements with clinical and molecular genetic data in Duchenne muscular dystrophy. AB - Clinical improvement following surgery in patients with Duchenne muscular dystrophy (DMD) may be influenced by the severity of muscle fiber damage. This study correlates morphometric alterations of muscle fiber types, severity of fat tissue proliferation and fibrosis with Western blots, multiplex polymerase chain reaction (PCR), and postoperative state in DMD. The main results of this study show that the mean diameter of type 2 fibers is usually markedly larger than that of type I fibers in DMD although the number of type 2 fibers is severely reduced. The mean percentage of the remaining type 1 fibers was in the range of 57-82%. The morphometric and histopathological results were in agreement with the clinically estimated postoperative state, especially in the patients who presented a severe state or suggestive clinical improvement. It is concluded that combination of both quantitative and qualitative evaluation of muscle biopsies is suitable for better evaluation of the postoperative state in patients with DMD, whereas severity of exon deletions correlated poorly with morphometry and postoperative clinical state. PMID- 10382910 TI - Severe clinical expression in X-linked Emery-Dreifuss muscular dystrophy. AB - X-linked Emery-Dreifuss muscular dystrophy (EDMD) is a relatively rare benign neuromuscular disorder which can vary remarkably in onset, course and severity. In the present study, a TCTAC deletion spanning the nucleotides 631-635 of the emerin gene caused an unusually severe disease phenotype including loss of ambulation and severe muscle wasting in two affected brothers. The same mutation has been reported previously in an unrelated family showing a significantly milder phenotype. The interfamilial heterogeneity in distribution and in severity of the features in the two families point to environmental or genetic modification as the cause of clinical variability in Emery-Dreifuss muscular dystrophy. PMID- 10382909 TI - Genotype-phenotype analysis in X-linked Emery-Dreifuss muscular dystrophy and identification of a missense mutation associated with a milder phenotype. AB - Direct sequencing of the emerin gene in 22 families with Emery-Dreifuss muscular dystrophy (EMD) revealed mutations in 21 (95%), confirming that emerin mutations can be identified in the majority of families with X-linked EMD. Most emerin mutations result in absence of the protein. In this study three mutations (a missense mutation Pro183Thr and two in-frame deletions removing residues 95-99 and 236-241, respectively) were unusual in being associated with expression of mutant protein. The phenotype in these families was compared in detail with the clinical features in cases with typical null mutations. For the in-frame deletions there were no significant differences. In the family with the missense mutation the phenotype was milder. Age at onset was later for first symptoms and for development of ankle contractures and muscle weakness. These findings have diagnostic implications as well as pointing to functionally important regions of the emerin protein. PMID- 10382911 TI - A new mutation in the myophosphorylase gene (Asn684Tyr) in a Spanish patient with McArdle's disease. AB - We have identified a novel missense mutation, an A-T transition at codon 684 in exon 17, changing an encoded asparagine to a tyrosine (Asn684Tyr) in a Spanish patient with typical McArdle's disease. The patient was a compound heterozygote, with a previously-described mutation (Gly204Ser) on the other allele. This report expands the molecular genetic heterogeneity in McArdle's disease, emphasizes the presence of private mutations in specific ethnic groups, and indicates that geographic origin must be considered before undertaking DNA analysis for diagnosis. PMID- 10382912 TI - McArdle's disease associated with homozygosity for the missense mutation Gly204Ser of the myophosphorylase gene in a Spanish patient. AB - We studied a pateint whose clinical, morphological and biochemical findings were consistent with McArdle's disease. Molecular genetic studies revealed that the patient did not harbor the common Arg49Stop mutation and was homozygous for the Gly204Ser mutation. Until now, no patient having the missense mutation in the two alleles has been documented. PMID- 10382913 TI - Lower limb surgery in Duchenne muscular dystrophy. AB - Two hundred and thirteen of 428 patients with Duchenne muscular dystrophy (DMD) of a prospective and open study were operated on bilaterally with hip and knee release, aponeurectomy of the iliotibial band and Achilles tendon lengthening. In 87 patients the operation was carried out during early restrictions of the lower limb joint mobility at an average age of 6.56 years (4.02-8.26, SD 1.42). The follow-up was on average 5.4 years (0.25-9.01, SD 2.7). This group was compared to a control group (natural history) consisting of 100 non-operated DMD patients. A significant (P < 0.001) release of the contractures could be obtained. Loss of walking ability occurred in the control group at an average of 9.29 years (5.85 13.63, SD 1.98) and in the operated group at an average of 10.55 years (8.17 14.39, SD 1.76). This shows that early lower limb surgery leads to a prolongation of independent ambulation of 1.25 years on average. In contrast to the patients of the control group all treated patients between ages 6 and 8 years could walk independently. The positive influence of early lower limb surgery could also be shown by the development of Hammersmith motor ability score, CIDD (Council of Investigation of Duchenne Dystrophy) grading and Vignos scale. Nevertheless, in consideration of the well-known course of DMD not only the prolongation of ambulation but also the achieved prolongation of assisted standing ability with no or mild contractures are aims of lower limb surgery. Since no improvement of muscle strength could be observed after lower limb surgery, further studies have to investigate if additionally administered steroids can prolong ambulation after early lower limb surgery. PMID- 10382914 TI - Effect of mexiletine on sea anemone toxin-induced non-inactivating sodium channels of rat skeletal muscle: a model of sodium channel myotonia. AB - The sea anemone toxin ATX II impairs skeletal muscle sodium channel inactivation, mimicking the persistent inward current observed in patients suffering from sodium channel myotonia. Mexiletine has beneficial effects on myotonia. To verify the efficiency of the drug on persistent inward current, we investigated the effect of 50 microM R(-)-mexiletine on sodium channels in cell-attached patches of rat skeletal muscle fibres, in the absence or presence of 2 microM ATX II. With the toxin, a proportion of channels displayed remarkable abnormal activity lasting the entire depolarisation, which resulted in a persistent inward current that represented up to 2.0% of the peak current. Mexiletine reduced by 75% the peak current elicited by depolarisation from -100 to -20 mV. This was due to the reduction by 60% of the maximal available peak current Imax and to the negative shift by -7 mV of steady-state inactivation. Mexiletine also greatly decreased the late current, but the effect was limited to 60% of reduction, comparable to that on Imax. Therefore mexiletine was able to block the ATX II-modified sodium channels, inhibiting the myotonia-producing persistent inward current. PMID- 10382915 TI - Molecular analysis of 4q35 rearrangements in fascioscapulohumeral muscular dystrophy (FSHD): application to family studies for a correct genetic advice and a reliable prenatal diagnosis of the disease. AB - In the majority of facioscapulohumeral muscular dystrophy (FSHD) families (about 95%) the genetic defect has been identified as a deletion of a variable number of KpnI repeats in the 4q35 region, although no specific transcripts from this locus have been isolated so far. Molecular diagnosis is based on the detection by probe p13E-11 of EcoRI small fragments, in the range 10-28 kb, that are resistant to BlnI digestion. In family studies this probe is used with other 4q35 polymorphic markers to assign the haplotype associated with the disease. So far, we performed DNA analysis in 145 FSHD families and identified the 4q35 DNA rearrangement not only in affected individuals, but also in healthy subjects at risk of transmitting the disease, such as non-penetrant gene carriers and somatic mosaics. In addition we applied prenatal tests to 19 fetuses, using DNA extracted from chorionic villi samples (CVS) at 10-11 weeks of gestation. The FSHD status, as determined by the presence of BlnI-resistant small fragments associated with the at risk haplotype, was assessed in nine fetuses; in the remaining 10 cases the disease was excluded. Our results show that molecular analysis of 4q35 rearrangements is a reliable indirect method to perform diagnostic, predictive and prenatal tests in FSHD. PMID- 10382916 TI - The Emery-Dreifuss Muscular Dystrophy Mutation Database. PMID- 10382917 TI - Neuromuscular disorders: gene location. PMID- 10382918 TI - Mitochondrial encephalomyopathies: gene mutation. PMID- 10382919 TI - Measurement in clinical disorder: from symptom onset to longitudinal outcome. PMID- 10382920 TI - Pain and somatosensory dysfunction in acute herpes zoster. AB - OBJECTIVE: To determine the nature of sensory change and its association with pain and allodynia in acute herpes zoster. DESIGN: Prospective clinical study. PATIENTS: One hundred thirteen immunocompetent patients with acute herpes zoster. METHODS: Onset, intensity, and quality of pain and severity of rash were recorded. Quantitative somatosensory testing for tactile and thermal thresholds, qualitative pinprick testing, and testing of dynamic and static allodynia were performed within the affected dermatome, its mirror-image dermatome, and in an adjacent dermatome bilaterally. RESULTS: Acute pain was reported as severe in 50%, moderate in 29%, mild in 12%, and absent in 9% of patients. Preherpetic pain (median = 4 days, range = 1-60 days) was experienced by 71%. Mechanical allodynia, dynamic, static, or both, was found in 37% of patients and was noted to extend one or more dermatomes outside the rash in 12%. In the affected dermatomes, thresholds were elevated for warmth and cold, lowered for heat pain, and unchanged for touch when compared with the contralateral side. Logistic regression analyses showed that compression-evoked allodynia, brush-evoked allodynia, and the history of preherpetic pain were more frequently encountered in patients with severe pain. Sensory threshold changes were not associated with the severity of pain or rash or with the presence of allodynia. CONCLUSION: Pain, allodynia, and altered sensation are common features of acute herpes zoster. They are likely to result primarily from widespread neural inflammation within the affected afferent system. The sensory changes found in acute herpes zoster are different from those reported in published studies on postherpetic neuralgia and suggest sensitization phenomena and preservation of tactile functions rather than major neural damage. The exact mechanisms for acute herpes zoster pain, however, remain speculative. PMID- 10382921 TI - Self-reported abuse history and pain complaints among young adults. AB - OBJECTIVE: Considerable evidence suggests that a self-reported history of physical and/or sexual abuse is more frequently reported among chronic pain populations and is associated with poorer adjustment to pain. However, previous research has typically included patients seeking treatment for pain, whereas few population-based studies have explored the association between abuse history and pain. This purpose of this study was to examine the association between self reported history of sexual or physical abuse and recent pain complaints, health related variables, and psychological disturbance among a nonclinical sample of young adults. DESIGN: Subjects were 426 (275 female, 151 male) college students who completed a series of questionnaires assessing abuse history, recent pain, health care utilization, perceived health, and psychological variables. RESULTS: Females reported a positive history of abuse (PHA) more frequently than males (43.5% vs. 23.8%), and females reported significantly higher rates for all types of abuse except physical abuse during childhood (p < 0.05). PHA subjects reported experiencing pain in more sites and pain of higher severity over the past month compared to subjects with a negative history of abuse (NHA) (p < 0.05). PHA subjects also reported more health care utilization and greater psychological disturbance, including depression, somatization, negative temperament, and higher levels of catastrophizing (p < 0.05). Interestingly, when somatization and depression scale scores were used as covariates, group differences in pain complaints and health care utilization became nonsignificant (p > 0.10). CONCLUSION: These findings suggest that a self-reported history of physical or sexual abuse is associated with increased pain complaints, health care utilization, and psychological disturbance even among young adults from a nonclinical population. Moreover, the association between abuse and pain complaints appears to be moderated at least in part by the higher levels of somatization and depression observed in the PHA group. PMID- 10382922 TI - Pain in nursing home residents: an exploration of prevalence, staff perspectives, and practical aspects of measurement. AB - OBJECTIVE: To help rectify the underdiagnosis of chronic pain in frail nursing home residents by developing a new feasible pain self-report instrument, the structured pain interview; to use this new tool to estimate pain prevalence and staff's knowledge of residents' pain in two nursing homes; and to compare the performance differences of the structured pain interview and the commonly used 0 10 scale. DESIGN: Cross-sectional survey. SETTING: One 120-bed VA-affiliated and one 125-bed university-affiliated, community-based nursing home in Durham, North Carolina. PATIENTS: One hundred fifty-eight chronic care nursing home residents without aphasia, acute illness, persistent vegetative status, or severe hearing impairment and 3 1 nursing home nurses. OUTCOME MEASURES: Pain prevalence according to resident self-report and nurse report; stability of response to the structured pain interview and 0-10 scale over 1 month; agreement between residents and nurses on the structured pain interview and 0-10 scale. RESULTS: Fifty-eight percent of the VA and 45% of the community nursing home residents reported pain. Forty-two percent at the VA and 20% at the community home were unable to respond to the 0-10 scale, compared with 7.5% and 14% using the structured pain interview. Stability of response to the structured pain interview at 1 month was 0.56 at the VA (nurse-resident agreement 0.38) and 0.72 in the community (nurse-resident agreement 0.07), which was very comparable to the 0-10 scale. CONCLUSIONS: We have developed a highly feasible tool for examining pain prevalence in nursing homes. This tool uncovered considerable miscommunication regarding pain between residents and staff. Improvement in pain communication between nursing home residents and staff is needed, so that more effective pain treatment programs can be developed for this vulnerable population. PMID- 10382923 TI - Validity of the dictionary of occupational titles residual functional capacity battery. AB - BACKGROUND DATA: The Dictionary of Occupational Titles (DOT) is a U.S. government publication that defines each job in the United States according to 20 job factors. Fishbain et al. (Spine 1994;19:872-80) developed a DOT residual functional capacity (RFC) battery whose predictive validity for employment/unemployment had not been tested previously. OBJECTIVES: The purposes of this study were as follows: (a) to determine whether results of a DOT-RFC battery performed at completion of pain facility treatment predicted employment status at 30 months' follow-up and (b) to determine whether the DOT-RFC battery predicted employment capacity as determined by the DOT employment levels of the chronic pain patients' (CPPs) jobs. STUDY DESIGN: This is a prospective low back pain CPP pain facility treatment study using employment status and the DOT occupational levels as outcome measures. METHODS: One hundred eighty-five consecutive CPPs who fitted the selection criteria completed a DOT-RFC battery at the completion of pain facility treatment and were contacted at 1, 3, 6, 12, 18, 24, and 30 months for determination of their employment status and DOT employment level. Eight DOT job factors plus pain and worker compensation status were found to be significantly different between employed and unemployed CPPs and between those employed in different DOT employment levels. For the 10 variables, stepwise discriminant analysis was used to select final predictor variables. Sensitivity and specificity were calculated along with pain level cutpoints that separated the groups. RESULTS: The eight DOT job factors found to be statistically significant between groups were the following: stooping, climbing, balancing, crouching, feeling shapes, handling left and right, lifting, carrying, and pain and worker compensation status. In the discriminant analysis, these variables could discriminate between the employed and unemployed categories, with a sensitivity and specificity of approximately 75%. The pain level cutpoint between employed and unemployed was 5.4 on a 10-point scale. CONCLUSIONS: We cannot as yet predict DOT-RFC employment levels. However, if a CPP can pass the above eight DOT job factors and has a pain level less than the 5.4 cutpoint, that CPP will have a 75% chance of being employed at 30 months after treatment at the pain facility. Therefore, some DOT-RFC battery job factors demonstrate a predictive validity in the "real work world." PMID- 10382924 TI - Empirical subgroups of the Coping Strategies Questionnaire-Revised: a multisample study. AB - OBJECTIVE: The purpose of this study was to examine the dimensions of coping, measured by the subscales of the new revised Coping Strategies Questionnaire (CSQ R) using factor analysis, and to perform cluster analysis on these factors to explore the existence of distinct subgroups. No published studies have identifed subgroups of chronic pain patients based on the use of CSQ coping strategies. SOURCE: A sample of 419 chronic low back pain patients from a multidisciplinary pain rehabilitation clinic and a sample of 556 chronic pain patients of mixed pain etiology presenting for treatment at an anesthesia pain clinic were used to establish reliability of factors and clusters. RESULTS: Both samples yielded very similar two-factor solutions, with initial solutions accounting for 67.1% and 69.1% of the total variance. The factors were characterized as cognitive coping and distraction. Three homogeneous subgroups were then identified that consisted of a group high on cognitive coping, a group with low overall ratings of response CSQ-R items in general, and a group with frequent endorsement of catastrophizing and distraction-related items. CONCLUSION: This paper is the first to report empirically derived subgroups from scores on the CSQ or CSQ-R. In addition, the three clusters were significantly different across measures of pain, psychological distress, and levels of physical functioning, demonstrating validity for the clusters. PMID- 10382925 TI - Toward more accurate use of the Beck Depression Inventory with chronic back pain patients. AB - OBJECTIVE: To improve the accuracy of the Beck Depression Inventory (BDI) in assessing depression in chronic back pain (CBP) patients, the pattern of depression symptomatology was evaluated. DESIGN: Analyses of BDI data obtained from nondepressed and depressed CBP patients were conducted to identify the major factors that differentiated among the patient groups. SETTING: CBP patients were recruited from a tertiary rehabilitation center. PATIENTS: One hundred one nondepressed and 99 depressed CBP patients from the tertiary treatment center. OUTCOME MEASURES AND RESULTS: Analyses of the BDI data revealed a general factor of depression severity that excluded items reflecting weight loss, sleep disturbance, and work inhibition. In addition, these analyses yielded a second factor reflecting somatic concerns and disability. Weight loss, sleep disturbance, and work inhibition failed to differentiate the depressed from the nondepressed CBP subjects, suggesting that these symptoms have poor diagnostic potential for CBP patients. CONCLUSIONS: This study demonstrated that the BDI can be used to generate important information about the severity of interference posed by pain on the functioning of an individual, while allowing for an independent evaluation of subjective indices of depression and somatic disturbances that need to be attended to by clinicians. PMID- 10382926 TI - Disease-specific and generic health outcomes: a model for the evaluation of long term intrathecal opioid therapy in noncancer low back pain patients. AB - OBJECTIVE: The present study provided comprehensive characterization of the long term outcomes of intrathecal opioid administration via a drug administration system (DAS) in chronic pain patients with predominantly low back pain. A conceptual framework based on multidimensional outcomes is proposed using both disease-specific and generic measures. DESIGN: Pre-post longitudinal data were collected in a retrospective fashion on 38 patients receiving intraspinal opioid therapy for a minimum of 36 months (average = 50 months). MAIN OUTCOME MEASURES: Disease-specific measures included magnitude of infused opioid, side effects/complications, pain ratings, McGill Pain Questionnaire, Beck Depression Inventory, Oswestry Disability Questionnaire, and patient estimated improvement in pain (0-100%). Generic measures of health included the Quality of Well-Being Scale, Medical Outcomes Study (MOS) Short Form 36 (SF-36), return to work, patient estimated improvement in functioning, overall patient satisfaction, and family opinion of patient improvement. RESULTS: Disease-specific outcomes. Patients receiving long-term intrathecal opioid administration showed a sixfold increase in morphine equivalents infusion rates across time. DAS patients showed a small but significant decrease in pain ratings from pre-treatment levels. Following 3 years or more of intrathecal opioid infusion, patients endorsed high pain levels on the McGill Pain Questionnaire, severe levels of disability via the Oswestry Disability Questionnaire, mild levels of depression based on the Beck Depression Inventory, and multiple side effects associated with the intrathecal opioids and complications related to the infusion system. On retrospective questioning, patients receiving long-term intrathecal opioid administration reported an average of 64% improvement in their pain and 48% improvement in functioning. Family members of patients reported that they observed on average a 61% improvement in patient's pain. Generic outcome measures. On the Quality of Well-Being Scale, patients reported significantly lower health-related quality of life than health maintenance organization enrollees with no known chronic condition and patients with rheumatoid arthritis (p < 0.001). On the MOS SF-36, patients reported significantly lower physical functioning than the U.S. general population, patients with uncomplicated medical conditions, diabetes-type II patients, and congestive heart failure patients. Mental functioning was comparable to the U.S. general population (p > 0.001). Fourteen percent of patients were working following implantation. Eighty-nine percent of patients reported good to excellent satisfaction with the long-term intrathecal opioid therapy. CONCLUSIONS: Results from this study revealed differences in findings across the outcome measures, highlighting the complexity of intrathecal opioid therapy. Generally, patients after 3 years or more of intrathecal opioid therapy can be characterized as hav ing substantially impaired physical functioning with a high prevalence of side effects. Despite poor physical functioning, patients endorsed relatively good mental health status with only mild depressive symptoms. Longitudinal pain ratings showed a modest decrease from pretreatment levels. On retrospective evaluation, patients and their family endorsed high levels of pain relief secondary to intrathecal therapy. Overall, findings support that intrathecal opioid therapy provides some therapeutic benefit although substantial physical impairment continues to cause debilitation in the patient population. PMID- 10382927 TI - Nerve root sleeve injections in patients with failed back surgery syndrome: a comparison of three solutions. AB - OBJECTIVE: To evaluate outcome in patients with failed back surgery syndrome treated with nerve root sleeve injections. DESIGN AND PATIENTS: An open, nonblinded, randomized study on 60 patients with documented fibrosis in fewer than three nerve roots. INTERVENTIONS: After random selection, 20 patients were injected with 1 ml bupivacaine 0.5% combined with 1500 units hyaluronidase and 1 ml saline per nerve root sleeve (group A), another 20 were treated with 1 ml bupivacaine 0.5% combined with 40 mg methylprednisolone solution (Depo Medrol) per nerve root (group B), and a third group was treated with bupivacaine 0.5% combined with 1500 units hyaluronidase and 40 mg methylprednisolone solution (group C). The volume of each injection was 2 ml. The injections were given twice at an interval of 1 week. OUTCOME MEASURES: The patients were evaluated on a verbal pain rating scale 1, 3, and 6 months after the second injection. The Kruskal-Wallis test was used to detect statistically significant differences among the three groups, and the analysis was refined with the Friedman test. RESULTS AND CONCLUSIONS: Overall, although injections induced analgesia at 1 month, these effects were reduced at 3- and 6-month follow-ups. No statistical differences were found between the three treatment groups (after 1 month, p = 0.71; after 3 months, p = 0.69; after 6 months, p = 0.66. The Friedman test showed a significant decrease in treatment score as a function of time in groups B and C (p = 0.015) but not in group A (p = 0.074). Corticosteroids seem responsible for the last phenomenon. PMID- 10382928 TI - Long-term opioid analgesic therapy for severe refractory lumbar spine pain. AB - OBJECTIVES: To determine the effect of opioid analgesics on pain and function in patients with severe, refractory low back pain and to see if any benefits were sustained long term. DESIGN: Longitudinal evaluation was conducted in two stages. Stage I was an opioid trial, and stage II was long-term treatment. Treated patients were compared with dropouts and trial failures. OUTCOME MEASURES: Pain was measured by the Numerical Rating Scale (NRS) and function was measured by the Oswestry Low Back Disability Score (OSW). Outcomes were evaluated by patient questionnaire and therefore not subject to investigator bias. SETTING: Private office practice. METHODS: Patients were treated for 6-12 weeks with a long-acting or sustained-release opioid. Those who improved significantly were treated long term. The treatment group was compared with dropouts and failures. RESULTS: Thirty-three patients underwent opioid trial. Treatment was discontinued because of intolerable side effects in 5 patients. In the remaining 28, mean NRS improved from 8.6 to 5.9 (p < 0.001), and mean OSW improved from 64 to 54 (p < 0.001). There were 21 patients treated long term (mean, 32 months). NRS improved from 8.45 to 4.90 (p < 0.001), and OSW improved from 64 to 50 (p < 0.001). Two patients returned to work. The changes in pain and function in the treatment group were significantly better than the comparison group. There was no drug diversion, addictive behavior, or organ toxicity. Doses remained stable. CONCLUSION: Long-term opioid analgesic therapy is reasonable treatment for some well-selected patients with refractory low back pain who have failed all other forms of care. PMID- 10382929 TI - Prediction of "intent", "discrepancy with intent", and "discrepancy with nonintent" for the patient with chronic pain to return to work after treatment at a pain facility. AB - OBJECTIVE: We previously determined that "intent" to return to work post pain facility treatment is the strongest predictor for actual return to work. The purposes of the present study were the following: to identify variables predicting "intent"; to predict membership in the "discrepant with intent" group [those chronic pain patients (CPPs) who do intend to return to work but do not]; and to predict membership in the "discrepant with nonintent" group (those CPPs who do not intend to return to work but do). DESIGN: A total of 128 CPPs completed a series of rating scales and yes/no questions relating to their preinjury job perceptions and a question relating to "intent" to return to the same type of preinjury job post-pain facility treatment. These CPPs were part of a grant study for prediction of return to work, and therefore their work status was determined at 1, 3, 6, 12, 18, 24, and 30 months posttreatment. Preinjury job perceptions and other demographic variables were utilized using stepwise discriminant analysis to identify variables predicting "intent" and predicting membership in the "discrepant with intent" and "discrepant with nonintent" groups. SETTING: Pain facility (multidisciplinary pain center). PATIENTS: Consecutive low back pain CPPs, mean age 41.66+/-9.54 years, with the most frequent highest educational status being high school completion (54.7%) and 60.2% being worker compensation CPPs. RESULTS: "Intent" was predicted by (in decreasing order of probability) postinjury job availability variables, job characteristic variables, and a litigation variable. "Discrepant with intent" was predicted by (in decreasing order of probability) for the 1-month follow-up time point, postinjury job availability variables, pain variables, a litigation variable, and a function perception variable, and for the final follow-up time point, pain variables only. "Discrepant with nonintent" was predicted by (in order of decreasing probability) for the 1-month follow-up time point, a job availability variable, a demographic variable, and a functional perception variable, and for the final follow-up time point a pain variable and a job availability variable. The percentage of CPPs correctly classified by each of these analyses was as follows: "intent" 81.25%, "discrepant with intent" 87.01% (at 1-month follow-up) and 74.03% (final follow-up), "discrepant with nonintent" 92.16% (at 1-month follow-up) and 75.00% (final follow-up). CONCLUSIONS: CPPs intentions of returning to their preinjury jobs are mainly determined by job availability and job characteristic variables but surprisingly not by pain variables. However, the results with "discrepant with intent" and "discrepant with nonintent" groups indicate that actual return to work is determined by an interaction between job availability variables and pain variables with pain variables predominating for long-term outcome. PMID- 10382930 TI - Proximal myofascial dysfunction in complex regional pain syndrome: a retrospective prevalence study. AB - OBJECTIVE: To assess the prevalence of clinically evident myofascial dysfunction (MD) and its relationship to motor neglect (MN) in Complex Regional Pain Syndrome (CRPS). DESIGN: Retrospective chart review. PATIENTS: Forty-one consecutively evaluated CRPS patients. OUTCOME MEASURES: (a) Prevalence of trigger points in the proximal musculature of the CRPS limb. (b) Prevalence of MN in a subset (n = 34). RESULTS: MD was detected in 61 % of CRPS patients. It was more prevalent in the upper limb (70%) than in the lower limb (47%). MN was more common in those who also had MD. CONCLUSION: MD is common in CRPS patients, especially in the upper limb and in those patients with MN. Prospective trials are needed to confirm these intriguing findings, which may have important implications regarding CRPS pathophysiology and treatment. PMID- 10382931 TI - Consensus report therapy guidelines for CRPS. PMID- 10382932 TI - Medullary and CNS dysfunction in chronic pain. PMID- 10382933 TI - Response to Gerald Aronoff. PMID- 10382934 TI - CRPS: are guidelines possible? PMID- 10382935 TI - Clinical use of rHuEPO in bone marrow transplantation. AB - Recipients of bone marrow (BMT) or peripheral blood progenitor stem cells (PBSCT) transplant have in the period following transplantation a frequent need for red blood cell transfusions and therefore an increased risk of blood-transmitted infections. The anaemia is caused mainly by myelosuppression after high-dose chemotherapy, but an impaired erythropoietin (EPO) production and an inappropriate EPO response may also contribute. Since recombinant human erythropoietin (rHuEPO) has been established as a treatment for renal anaemia it has been of interest whether treatment may be of benefit in the transplant setting. This paper gives an overview of the studies conducted to date with rHuEPO treatment in patients receiving bone marrow transplants. Current data do not support any transfusional benefits when rHuEPO is used in patients receiving autologous transplants. However, in patients receiving allogeneic transplants several studies clearly indicate a therapeutic role for rHuEPO with patients showing accelerated erythroid engraftment, increased haemoglobin levels, a reduced requirement for red blood cell transfusions, and a shortened time to transfusion independence. Especially patients with immune haemolysis after transplantation seems to benefit from the treatment. In addition, rHuEPO treatment has been used for late-onset anaemia after BMT and to prevent the need for homologous red blood cell transfusions to the BMT donor. To reduce costs, it is important in future studies to identify not only the optimal dose and route of administration of rHuEPO but also the most effective combination with other haematopoietic growth factors and cytokines that are used before and after transplantation. PMID- 10382937 TI - Will modern cancer vaccines reach clinical practice? PMID- 10382936 TI - Lymphoid disorders associated with HHV-8/KSHV infection: facts and contentions. AB - Following the demonstration in 1994, that Kaposi's sarcoma (KS) was associated with a novel virus (KSHV or HHV-8) belonging to the lymphotropic herpes family, this virus was also found in certain lymphoid neoplasias of immunodeficient (HIV+) and immune competent hosts. The association of HHV-8/KSHV infection is now well established with primary effusion lymphoma (PEL) or body cavity based lymphoma (BCBL) and multicentric Castleman's disease (MCD) of the plasma cell type. A possible pathogenic role of HHV-8/KSHV in other lymphoid tumours including primary central nervous system lymphoma (PCNSL) and multiple myeloma (MM) as well as some atypical lymphoproliferations and sarcoidosis has also been suggested, but this is at present a controversial matter, or not confirmed. Several HHV-8/KSHV genes, including potential oncogenes, genes homologous to various cellular genes and growth factors have been incriminated in the pathogenesis of KS and PEL/BCBL, but a common pathogenic mechanism for the clearly diverse proliferations represented by PEL, MCD and KS is at present not evident. PMID- 10382938 TI - Tumour cell detection in G-CSF mobilised stem cell harvests of patients with breast cancer. AB - Peripheral blood stem cells were mobilised with G-CSF from steady-state haemopoiesis after previous anthracyclin-containing standard dose chemotherapy in patients with high-risk breast cancer. 48 samples were obtained from patients with stage II-III breast cancer and > or = 10 lymph nodes, 15 samples from patients with chemotherapy sensitive metastatic disease, and 13 samples from women with inflammatory breast cancer. 44 samples were first or single leukaphereses and 32 samples were second or third harvests. Aliquots were searched for contaminating tumour cells by immunocytochemistry (IC) and cytokeratin-19 reverse transcriptase polymerase chain reaction rtPCR). The median count of MNCs examined by IC was 2 x 10(6); cDNA prepared from 2 x 10(7) cells was subjected to PCR. Fifty-nine samples were examined by immunocytochemistry, 36 samples by rtPCR, and 19 samples by both techniques. Samples investigated by IC and rtPCR were judged as positive if there was at least one positive test. On the whole, 42/79 (55.3%) of the samples were positive with an insignificant trend to a higher positivity rate in second or subsequent leukaphereses (52.3% vs 59.3%). The median tumour cell load per 10(6) MNCs was low with 0.5 (0-7) cells in all, and a total of 2.2 (0.5-7) cells in positive specimen. Differences in the cancer cell load of first and subsequent leukaphereses and between subgroups of patients were not found. PCR and IC gave consistent results in 63.2%. This phenomenon can be explained by the greater sensitivity of the molecular method and by a Poisson distribution of coharvested tumour cells in samples. Tumour cell contamination in G-CSF mobilised stem cells from patients with breast cancer from steady state haemopoiesis after preceding anthacyclin-containing chemotherapy is frequent, but the tumour cell load is low. To allow a comparison of different studies dealing with cancer cell contamination in stem cells, standardisation of assays is necessary. PMID- 10382940 TI - Serum level of cytokeratin 19 fragment (CYFRA 21-1) indicates tumour stage and prognosis of squamous cell carcinoma of the oesophagus. AB - To determine the clinical efficacy of serum concentration of cytokeratin 19 fragment (CYFRA 21-1), sera from 66 patients with oesophageal squamous cell carcinoma were examined, and 54 surgically resected specimens were immunohistochemically stained for cytokeratin 19 (CK-19). The patients with positive CK-19 staining in the tissues of their carcinomas had significantly higher serum CYFRA 21-1 levels compared with those with negative CK-19 staining. When the cut-off value was defined as 2.0 ng/mL, CYFRA 21-1 had a higher positive ratio than that of either squamous cell carcinoma antigen (SCC-Ag) or carcinoembryonic antigen (CEA). Serum CYFRA 21-1 level increased significantly along with the clinical stages. In addition, serum CYFRA 21-1 level served as a prognostic factor for patients with oesophageal carcinoma after surgery, whilst SSC-Ag and CEA is not connected with the outcome. These findings suggest that the serum CYFRA 21-1 probably originated from the tumour tissue is an important marker for determining the stage and outcome of oesophageal carcinoma. PMID- 10382939 TI - Expression of p53, p21/waf, bcl-2, bax, Rb and Ki67 proteins in colorectal adenocarcinomas. AB - This study investigated the combined immunoexpression of p53, p21, bcl-2, bax, Rb and Ki67 proteins in colorectal adenocarcinomas and correlated expression patterns with tumour stage and grade. Paraffin sections from 98 cases of colorectal adenocarcinomas were stained by immunohistochemistry for p53, p21, bcl 2, bax, Rb and MIB-1 (Ki67) proteins. In addition, 12 cases of colorectal adenomas and normal colorectal mucosa were studied in parallel. P53, p21, bcl-2, bax, Rb and Ki67 proteins were detected in at least 5% of tumour cells in 63/98, 72/98, 52/98, 96/98 and 98/98 adenocarcinomas, respectively. Comparative study of the normal-adenoma-carcinoma tissues revealed abrogation of the normal immunotopography in adenomas and adenocarcinomas, and considerable modifications, increase or reduction, of the expression of p53, p21, bcl-2, bax, Rb and Ki67 proteins in adenocarcinomas when compared with normal mucosa and adenomas. Statistically significant correlations were found between low bax expression and Dukes C stage of carcinomas, Ki67 expression and carcinoma grade, and Ki67 and Rb expression. P53, p21, bcl-2 and Rb immunoexpression did not correlate with tumour stage or grade. Our findings show that low bax immunoexpression is frequently related to colorectal adenocarcinomas with lymph node metastases suggesting that low levels of bax expression play a role in late stage colorectal cancer. The correlation between Ki67 and Rb expression, in view of previous data that the hyperphosphorylated inactive Rb protein is frequently increased in colorectal adenocarcinomas, suggests that Rb protein is somewhat ineffective in inhibiting the cell-cycle progression in these malignancies. Furthermore, our findings provide immunohistochemical evidence that the abrogation of the normal immunotopography and the modifications of the expression of p53, p21, bcl-2, bax, Rb and Ki67 proteins reflect important events in colorectal oncogenesis. PMID- 10382941 TI - Linker-based GnRH-PE chimeric proteins inhibit cancer growth in nude mice. AB - Since the number of cancer-related deaths has not decreased in recent years, major efforts are being made to find new drugs for cancer treatment. In this report we introduce the gonadotropin releasing hormone-Pseudomonas exotoxin (GnRH PE) based chimeric proteins L-GnRH-PE66 and L-GnRH-PE40. These proteins are composed of a GnRH moiety attached to modified forms of Pseudomonas exotoxin via a polylinker (gly4ser)2. The chimeric proteins L-GnRH-PE66 and L-GnRH-PE40 have the ability to target and kill adenocarcinoma cell lines in vitro, whereas non adenocarcinoma cell lines are not affected. We demonstrate that L-GnRH-PE66 and L GnRH-PE40 efficiently inhibit cancer growth. Nude mice were injected subcutaneously with the SW-48 adenocarcinoma cell line to produce xenograft tumours. When the tumours were established and visible, the animals were injected with chimeric proteins for 10 days. At the end of this period, a reduction of up to 3-fold in tumor size was obtained in the treated mice, as compared with the control group, which received equivalent amounts of GnRH; the difference was even greater 13 days after termination of treatment. Thus, the chimeric proteins L GnRH-PE66 and L-GnRH-PE40 are promising candidates for treatment of a variety of adenocarcinomas and their use in humans should be considered. PMID- 10382942 TI - Role of macrophage colony-stimulating factor in the differentiation and expansion of monocytes and dendritic cells from CD34+ progenitor cells. AB - The present study focused on whether it is possible to expand monocytic cells from CD34+ progenitor cells by using macrophage colony-stimulating factor (M-CSF) in the absence and presence of mast cell growth factor (MGF) and IL-6. It was demonstrated that CD34+ cells differentiate without expansion to functional mature monocytic cells in the presence of M-CSF or combinations of M-CSF plus IL 6 and MGF. A different response pattern was observed for the number of clonogenic cells. The addition of IL-6 or both IL-6 and MGF to M-CSF containing cultures resulted in significant higher numbers of colony-forming unit-macrophage (CFU-M) as tested in clonogenic and 3H-thymidine assays. Furthermore, M-CSF plus both IL 6 and MGF appeared to be the most potent combination to preserve the monocytic precursor in cell suspension culture assays. These results indicate that IL-6 and MGF in conjunction with M-CSF affect CD34+ cells especially at precursor level without distinct effect on the more mature stages. Secondly we studied whether M CSF is only critical for the monocytic lineage or also affects dendritic cell (DC) development. Indeed, we were able to culture CD83+ DC from CD34+ progenitor cells in the presence of M-CSF in conjunction with TNF-alpha, IL-4, and MGF although their absolute number is almost threefold lower than the number of CD83+ cells yielded from GM-CSF plus TNF-alpha, IL-4, and MGF stimulated CD34+ cells. PMID- 10382943 TI - Influence of perioperative whole blood transfusions on lymphocyte subpopulations in patients with stage II breast cancer. AB - Preliminary reports have suggested an adverse relationship between blood transfusion and survival after surgery in patients with solid tumour. One might postulate that from these studies, perioperative blood transfusions alter host immune defences. We therefore examined the influence of homologous whole blood transfusion on circulating lymphocyte subpopulations in transfused patients compared with non-transfused patients. Fifty-one women with Stage II breast cancer who underwent surgical procedures were studied. Patients were classified into two groups on the basis of whether or not they had received blood transfusion. The lymphocyte subpopulations were analyzed by flow cytometry before cancer surgery and three weeks after the operation. CD3+, CD4+, CD8+, and CD20+ cells as the lymphocyte subsets were quantitated using appropriate monoclonal antibodies. No significant differences between pre- and postoperative lymphocyte subset levels were seen in non-transfused patients. However, there was a statistically significant increase in the CD8+ cell count; decreasing CD4+ cell count and decreased CD3+ cells levels were observed in the transfused group (P < 0.05). Although these early results of the study suggest that the blood transfusions could be associated with alterations in lymphocyte populations, additional studies are needed to elucidate the possible mechanism of the transfusion-induced immunological modulations. PMID- 10382944 TI - Arsenic trioxide induces apoptosis of myeloid leukemia cells by activation of caspases. AB - The primary objective of this study was to determine whether caspases are involved in arsenic trioxide(ATO)-induced apoptosis of human myeloid leukemia cells. A secondary objective was to determine whether apoptosis induced by ATO compared with VP-16 is differentially affected by an activator of protein kinase C (PKC), phorbol 12-myristate 13-acetate (PMA), which has been reported to inhibit apoptosis induced by some chemotherapeutic agents. NB4 and HL60 cells were incubated with ATO in the presence and absence of the caspase protease inhibitors Z-VAD.fmk or Y-VAD.cho. Apoptosis was assessed by morphology, DNA laddering and flow cytometry. Poly (ADP-ribose) polymerase (PARP) cleavage was used as a marker for the activation of caspases. PARP cleavage occurred during ATO-induced apoptosis in both NB4 and HL60 cells. Z-VAD.fmk, a broad-spectrum inhibitor, could block ATO-induced apoptosis and PARP cleavage, whilst Y-VAD.cho, a selective inhibitor of caspase 1, had no such effect. PMA pre-incubation for up to 8 hours under conditions known to activate PKC had no effect on either ATO- or VP-16-induced apoptosis. We conclude that in cultured myeloid leukemia cells ATO induced apoptosis is executed by caspases from the distal, PARP-cleaving part of the activation cascade and that PKC activation has no effect on apoptosis induced by either ATO or VP-16 in these cells. PMID- 10382946 TI - Therapeutic efficacy of theophylline in chronic lymphocytic leukemia. AB - Theophylline, a methylxanthine commonly used as a treatment for asthma, has been shown to induce apoptosis in chronic lymphocytic leukemia (CLL) cells both in vitro and in vivo. We have treated three advanced CLL patients with theophylline, and seen responses in two. The clinical courses of the responders are presented, and the literature concerning theophylline as therapy for CLL is reviewed. PMID- 10382945 TI - Chronic lymphocytic leukaemia presenting with central nervous system involvement. AB - A 68-year-old man presented with hemiparesis, lymphocytosis, and cerebral lesions on MRI. Flow cytometry of blood, bone marrow and cerebrospinal fluid showed B-CLL lymphocytes with bright CD20 expression, sIg, and absence of CD23 antigen. Fluorescence in situ hybridisation showed trisomy 12 in 50% of analysed peripheral mononuclear cells. The patient died 6 months after the diagnosis. Rapidly progressive and fatal course of the disease was consistent with known bad prognostic significance of CD20 bright expression and trisomy 12. PMID- 10382947 TI - Cyclophosphamide, cytosine arabinoside and TBI as a conditioning regimen for allogeneic bone marrow transplantation in patients with leukemia. AB - This is a prospective study designed to determine the toxicity, efficacy and antileukemic effect of high-dose cytosine arabinoside (ara-C), cyclophosphamide and total body irradiation (TBI) as a myeloablative regimen prior to allogeneic bone marrow transplantation for patients with hematologic malignancies. Fifty eight patients with hematologic malignancies were treated with cyclophosphamide, high-dose ara-C and total body irradiation (TBI) followed by allogeneic bone marrow transplantation. Fifty patients had good prognosis disease and eight had poor prognosis disease. Cyclosporine and short-course methotrexate were used for graft-versus-host disease (GVHD) prophylaxis. The conditioning regimen consisted of ara-C 3000 mg/m2 twice a day x six doses on days -7, -6, and -5; cyclophosphamide 1800 mg/m2 on days -4 and -3; and TBI 1400 cGy midline dose at 5 cGy/min in eight total fractions administered twice a day on days -4, -3, -2, and -1. The bone marrow was infused on day 0 (zero). Toxicity related to the conditioning regimen was comparable to that reported with other conditioning regimens, except for diarrhea which appears to be more frequent. The actuarial survival at 1 year was 69% (58-82) and at 5 years was 54% (42-69) with the numbers in parentheses representing the 95% confidence interval of the Kaplan Meier estimate. After a median follow-up of 28 months, 31 of 58 (53%) patients are alive without evidence of disease. Only four of the 58 patients (7%) have relapsed. Cyclophosphamide, ara-C and TBI is a safe and effective myeloablative regimen for patients with leukemia. The overall relapse rate in our study was 7% with a median follow-up of 28 months and appears to be lower than relapse rates reported in other series. This is probably due to the added antileukemic effect of ara-C. This regimen should be compared with other myeloablative regimens in a controlled study. PMID- 10382948 TI - Mobilization of peripheral blood progenitor cells (PBPC) in patients undergoing chemotherapy followed by autologous peripheral blood stem cell transplant (SCT) for high risk breast cancer (HRBC). AB - We have determined the effect of delayed addition of G-CSF after chemotherapy on PBPC mobilization in a group of 30 patients with high risk breast cancer (HRBC) undergoing standard chemotherapy followed by high-dose chemotherapy (HDCT) and autologous SCT. Patients received FAC chemotherapy every 21 days followed by G CSF at doses of 5 microg/kg/day starting on day +15 (groups 1 and 2) or +8 (group 3) after chemotherapy. PBPC collections were performed daily starting after 4 doses of G-CSF and continued until more than 2.5 x 10(6) CD34+ cells had been collected. In group 1, steady-state BM progenitors were also harvested and used for SCT. Groups 2 and 3 received PBPC only. The median number of collections was three in each group. Significantly more PB CD34+ cells were collected in patients receiving G-CSF starting on day 8 vs day 15 (9.43 x 10(6)/kg and 6.2 x 10(6)/kg, respectively) (P < 0.05). After conditioning chemotherapy all harvested cells including BM and PBPC were reinfused. Neutrophil and platelet engraftment was significantly faster in patients transplanted with day 8 G-CSF-mobilized PBPC (P < 0.05) and was associated with lower transplant related morbidity as reflected by days of fever, antibiotics or hospitalization (P < 0.05). Both schedules of mobilization provided successful long-term engraftment with 1 year post transplant counts above 80% of pretransplant values. In conclusion, we demonstrate that delayed addition of G-CSF results in successful mobilization and collection of PBPC with significant advantage of day 8 G-CSF vs day 15. PBPC collections can be scheduled on a fixed day instead of being guided by the PB counts which provides a practical advantage. Transplantation of such progenitors results in rapid short-term and long-term trilineage engraftment. PMID- 10382949 TI - Culturing human umbilical cord blood: a comparison of mononuclear vs CD34+ selected cells. AB - We compared UCB mononuclear cells (MNC) with CD34+ selected cells in a serum-free static culture system. Cell number proliferation of MNCs was inferior to CD34+ selected cells. MNCs, however, showed a substantial increase from 0.94% CD34+ cells on day 0 to 5.8% on day 7, whereas in the CD34+ selected samples the CD34+ cell content declined continously from 62.2% on day 0 to 27.7% on day 7. The number of CFU-GM increased during culture of both cell fractions. Here, only the MNCs showed a substantial increase in clonogenicity on day 7 and day 14 to 11.1- and 4.1-fold input, respectively. This expansion of the CD34+ progenitor cell pool in the MNCs fraction was at least in part attributable to T cells, since the physical abrogation of T cells blocked this effect. Refeeding and reseeding of cells on day 7 had stimulating effects especially on the CD34+ cells, where cell number proliferation increased from 16.3-fold without to 58.1-fold on day 14. Also, we could find sporadic chromosomal aberrations in four of 100 metaphases examined after 7-20 days of ex vivo expansion. The significance of this observation needs to be clarified in a larger series. PMID- 10382950 TI - Circulating basic fibroblast growth factor declines during Cy/TBI bone marrow transplantation. AB - Basic fibroblast growth factor (bFGF) inhibits radiation-induced apoptosis, and radioprotects haematopoietic, cartilage growth plate, pulmonary and gastrointestinal tissues. Conversely, chronic overexpression of bFGF may promote fibrosis. We measured the endogenous circulating bFGF in blood of patients undergoing conditioning TBI. Twenty-six patients with haematopoietic malignancies were conditioned with cyclophosphamide/TBI for allogeneic BMT. Daily blood samples were collected each morning prior to, during, and for several days after TBI. bFGF levels in plasma of normal volunteers are 0.8-26 pg/ml. bFGF was below detectability in 22%, 30% and 45% of patients pre-TBI, during TBI or post-TBI respectively. Mean circulating plasma levels of bFGF decreased from a median of 52 pg/ml pre-TBI to 26 pg/ml during TBI, and to 5 pg/ml post-TBI. Among the 26 patients, 13 had more than one non-detectable plasma bFGF level, an additional five had at least one non-detectable level, and only eight patients had detectable levels in all daily samples. Naturally high levels of bFGF were observed in some patients undergoing fractionated TBI. In contrast, as many as 79% of patients had low bFGF levels in one or more samples. The impact of endogenous bFGF on the tolerance of normal tissues to irradiation is unknown, and warrants further study. PMID- 10382951 TI - High spontaneous IL-10 production in unrelated bone marrow transplant recipients is associated with fewer transplant-related complications and early deaths. AB - Interleukin 10 (IL-10) is a potent inhibitor of proliferative T cell responses toward alloantigens, and suppresses the production of pro-inflammatory cytokines which are important in cellular activation and recruitment to sites of inflammation. Because of these properties, we hypothesized that high IL-10 production in patients prior to BMT may predict a better outcome. To investigate this, peripheral blood mononuclear cells (PBMNC) were obtained from 58 recipients (11 autologous, 25 related donor (RD), and 22 unrelated donor (URD)), prior to conditioning therapy. PBMNC were cultured for 24 h in the presence and absence of lipopolysaccharide (LPS) and culture supernatants were assayed for IL-10 using an ELISA method. Spontaneously produced and LPS-stimulated IL-10 levels were correlated with the development of transplant-related complications (TRC) including grade II-IV acute GVHD, veno-occlusive disease, idiopathic pneumonia syndrome and multi-organ dysfunction syndrome, and with death before day 100. For the autologous group, there were no TRC and only one death prior to day 100; therefore, no statistical comparisons to IL-10 levels could be made. In the RD group, 36% developed one or more TRC and 24% died before day 100; however, there were no statistically significant associations between spontaneous or LPS-induced IL-10 levels. In URD patients 41% developed TRC and 55% died prior to day 100. In this group, higher levels of spontaneous IL-10 production were associated with a lower overall occurrence of TRC (P = 0.03) and early death (P = 0.04). Our data would indicate that higher levels of IL-10 production prior to URD BMT may predict fewer TRC, as well as early deaths. The hypothesis that high IL-10 production prior to BMT may decrease complications following URD BMT warrants further testing. PMID- 10382952 TI - Regimen-related toxicity and non-relapse mortality with high-dose cyclophosphamide, carmustine (BCNU) and etoposide (VP16-213) (CBV) and CBV plus cisplatin (CBVP) followed by autologous stem cell transplantation in patients with Hodgkin's disease. AB - This analysis compares the regimen-related toxicity (RRT) and overall non-relapse mortality (NRM) in Hodgkin's disease patients conditioned with either CBV (cyclophosphamide, BCNU (carmustine), and VP16-213 (etoposide)) (26 patients) or CBVP (CBV + cisplatin) (68 patients) followed by autologous stem cell transplantation (ASCT). CBVP included a continuous infusion rather than intermittent doses of etoposide, a lower BCNU dose and the addition of cisplatin. RRT and NRM were determined for each regimen and compared; risk factors for each were examined by multivariate analysis. Grade IV (fatal) RRT occurred in five patients (pulmonary in two, cardiac in two, and central nervous system in one). Eighteen patients experienced grade II-III pulmonary RRT, consistent with BCNU damage in 15. Prior nitrosourea exposure was the main risk factor for pulmonary RRT. Grade II mucosal and hepatic RRT occurred less often after CBVP vs CBV (P = 0.031 and 0.0003, respectively). In addition, three other early and eight late non-relapse deaths were seen. Median follow-up of the entire group is 5.1 (range 2.8-10.2) years. The probability of overall NRM was 26% (95% confidence interval (CI) 13-50%) with CBV vs 23% (95% CI 12-41%) with CBVP (P = 0.40). The progression-free survival and relapse rates were similar. Although the rates of fatal RRT, pulmonary RRT and overall NRM were similar with CBV or CBVP, CBVP produced less mucosal and liver RRT with a comparable antitumor effect. As many autografted patients are cured, future efforts should include measures to decrease NRM. PMID- 10382953 TI - Prevention and treatment of graft-versus-host disease by down-regulation of anti host reactivity with veto cells of host origin. AB - Control of graft-versus-host disease (GVHD) post allogeneic spleen cell transplantation was partly achieved by administration of host-type lymphocytes. (C57BL/6 x BALB/c) F1 mice were conditioned by total body irradiation and transplanted with parental C57BL/6 spleen cells to induce severe GVHD. Infusion of mature host-type lymphocytes did not change the degree of chimerism but protected most of the recipients from lethal GVHD. Treatment of host-type blood cells with anti-CD8 antibodies resulted in loss of the GVHD protective effects mediated by host cells, whereas after treatment of the host with anti-CD4 antibodies protective host cells given 4 days after induction of GVHD resulted in successful rescue of all transplanted recipients from lethal GVHD. Administration of host blood cells could effectively protect against GVHD induced by donor spleen cells and low-dose rIL-2. GVHD protection resulted from down-regulation of alloreactive donor T cells and not by rejection of GVHD effector cells. Taken together, administration of host hematopoietic cells particularly host CD8+ T cells, may be considered for prevention or control of GVHD following allogeneic bone marrow transplantation, thus also explaining the increased resistance of mixed chimeras to GVHD. PMID- 10382954 TI - Pre-treatment of transplant bone marrow cells with hydrocortisone and cyclosporin A alleviates graft-versus-host reaction in a murine allogeneic host-donor combination. AB - The aim of the study was to alleviate graft-versus-host reaction (GVHR) by pre treatment of the bone marrow (BM) transplant with hydrocortisone (HC) and cyclosporin A (CsA) in C57BL/6J (donor) --> CBA/J (recipient) mouse combination. BM cells were exposed to HC and CsA for 1 h at 37 degrees C and then injected into lethally irradiated (9.5 Gy) mice at a dose of 2 x 10(6) BM cells/mouse. Haematopoietic recovery was assessed on day 12, and survival was followed for 100 days. Combinations of 1000 microg/ml HC and 100 microg/ml CsA, and 100 microg/ml HC and 10 microg/ml CsA significantly reduced MLR and additively mitigated GVHR in vivo, achieving 40% and 26% survival rates, respectively. However, HC and CsA altered neither the peripheral blood cell counts nor in vitro and in vivo BM cell clonogenic potential. Additional studies have shown that HC and CsA blocked con A driven differentiation of CD8+ and CD4+ CD8+ lymph node cells (LNC) and progression of LNC to S + G2/M cell cycle phases, and inhibited IL-1, IL-2 and TGF-beta while enhancing GM-CSF gene expression in BM cells. Taken together, these data indicate that the pre-treatment of the BM transplant with HC and CsA results in inactivation of GVHR effector cells and mitigation of GVHR while sparing BM repopulating capacity. PMID- 10382955 TI - Recognition of leukemic blasts by HLA-DPB1-specific cytotoxic T cell clones: a perspective for adjuvant immunotherapy post-bone marrow transplantation. AB - There is increasing evidence that the immune response plays a role in the prevention of leukemic relapses after allogeneic bone marrow transplantation (BMT). Producing this effect (referred to as the graft-versus-leukemia reaction or GVL) is a current goal of clinical transplantation. At present, all protocols rely on the injection of donor T cells with unknown specificities. In keeping with this approach, we recently proposed the use of a single allogeneic T cell clone transfected with the HSv-tk gene to target an HLA-DPB1 mismatch in the GVH direction. For this strategy to be successful, HLA-DP antigens must be expressed on leukemic cells, which should be recognised by the HLA-DP-specific T cell clone and subsequently destroyed. In the present study, differential expression of HLA DR, -DQ and -DP was tested by fluorescence using monoclonal antibodies on a panel of 46 acute myeloid leukemias (AML), 28 acute lymphoblastic leukemias (ALL) and 31 chronic lymphocytic leukemias of B cell origin (B-CLL). The vast majority of leukemic cells expressed HLA-DP antigens although with considerable variability. HLA-DPB1 genotyped leukemic cells were used as target cells for an HLA-DPB1*0401 specific T cell clone. Specific recognition of leukemic blasts was demonstrated for 11 out of 11 B-CLL, 11 out of 19 AML and nine out of 16 ALL. These data show that most leukemic blasts are accessible to direct lysis by allogeneic HLA-DP specific T cells. PMID- 10382956 TI - Myelodysplasia and acute leukemia following high-dose chemotherapy and autologous bone marrow or peripheral blood stem cell transplantation. AB - Therapy-related myelodysplastic syndrome (t-MDS)/acute myeloid leukemia (t-AML) has been reported after autologous bone marrow or peripheral blood stem cell transplantation (ABMT/PBSCT) for various malignancies. We retrospectively reviewed all adult ABMT/PBSCT cases performed at the University of Chicago Medical Center from 1985 to 1997 in order to determine the incidence of therapy related leukemia. Among 649 patients, seven (1.1%) developed therapy-related acute lymphoblastic leukemia (one patient) or t-MDS/t-AML (six patients). Of these seven, primary malignancies included one case of breast carcinoma, five cases of Hodgkin's disease (HD) and one case of non-Hodgkin's lymphoma (NHL). Disease-specific incidences for therapy-related leukemia occurring after ABMT/PBSCT were one in 354 (0.3%) for breast carcinoma, five in 79 (6.3%) for HD and one in 103 (1%) for NHL. The median latency periods for the development of therapy-related leukemia from the time of initial diagnosis and of ABMT/PBSCT were 5.5 and 1.5 years, respectively, for the combined HD and NHL group of patients and 4.4 and 2.8 years, respectively, for the one breast carcinoma patient. All seven patients had clonal cytogenetic abnormalities, and five had recurring abnormalities typical of myeloid disorders. Given the similar latency period observed in patients treated with conventional chemotherapy alone, our findings support the hypothesis that therapy-related leukemia after ABMT/PBSCT likely results from pre-transplant therapy. Early detection of therapy-related leukemia is therefore critical to exclude these patients from undergoing ABMT/PBSCT. PMID- 10382957 TI - Opportunistic CNS infection after bone marrow transplantation. AB - We retrospectively identified opportunistic CNS infections in 655 patients who had undergone allogeneic, syngeneic or autologous BMT or PBSCT between 1990 and 1997. Twenty-seven patients (4%) developed CNS infections. All CNS infections occurred in allogeneic BMT or PBSCT patients. The most common CNS infections were toxoplasma encephalitis (74%) and cerebral aspergillosis (18%). Furthermore, we identified one patient with candida encephalitis and one patient with viral encephalitis. Overall mortality of patients with opportunistic CNS infection was 67%. There were two different groups of toxoplasma encephalitis with a different appearance on MR imaging. The first group showed edema, but no gadolinium enhancement, whereas the second group exhibited typical MRI appearances with the exception of frequent hemorrhagic transformation. The first group had a significant shorter latency between BMT and onset of CNS infection (mean 45 days vs 180 days, P = 0.02), a significant higher daily dose of corticosteroids as treatment for graft-versus-host disease (GVHD) (P = 0.01), more severe GVHD and a higher mortality (71% vs 36%). This study shows that the most common CNS infections in our patient population are toxoplasma encephalitis and cerebral aspergillosis, that there are two distinct subgroups of toxoplasma encephalitis and that CNS infections occur after allogeneic BMT only. PMID- 10382958 TI - Using at least 5x10(6)/kg CD34+ cells for autologous stem cell transplantation significantly reduces febrile complications and use of antibiotics after transplantation. AB - For autologous stem cell transplantation, it is common practice to infuse at least 2 x 10(6)/kg CD34+ cells to ensure rapid engraftment. However it was recently claimed that increasing the threshold to 5 x 10(6)/kg leads to a faster platelet engraftment. To evaluate these threshold values in our patient population we undertook a retrospective analysis of 127 autologous transplants performed at our institution between 1992 and 1998. Diagnoses included Hodgkin's and non-Hodgkin's lymphoma, myeloma, acute leukaemias and solid tumours. The transplant was peripheral blood stem cells in 107 cases and CD34-selected peripheral blood stem cells in 20 cases. The median number of transplanted CD34+ cells was 3.2 x 10(6)/kg (range 0.64-25.9 x 10(6)/kg). Haematopoietic recovery to a neutrophil count >0.5 x 10(9)/l took a median of 10 (range 5-16) days from transplant. When comparing patients receiving at least 5 x 10(6)/kg and 2-5 x 10(6)/kg CD34+ cells we found a significant reduction in the median number of days with fever (1 vs 3.5 days, P = 0.0025), incidence of fever (78.8 vs 92.1%, P = 0.032) as well as duration of antibiotic treatment (7 vs 10 days, P = 0.038). This was paralleled by a faster neutrophil recovery to 0.5 x 10(9)/l (9 vs 10 days, P = 0.047). There was no significant difference in the number of platelet or red cell transfusions between the two groups. We conclude that transplantation with a stem cell dose of at least 5 x 10(6)/kg CD34+ cells reduces infectious complications and should thereby increase the safety of this type of therapy while reducing duration (and cost) of antibiotic therapy. The transplantation threshold should thus not remain at 2 x 10(6)/kg particularly in patients with a good stem cell mobilisation capacity. PMID- 10382959 TI - A locus on chromosome 2 influences the development of acute graft-versus-host disease in a murine model. AB - Despite contemporary typing procedures for bone marrow transplantation (BMT), graft-versus-host disease (GVHD) continues to be a major complication of transplants performed between MHC-matched donors and recipients. Although GVHD can be alleviated by T cell depletion, this procedure increases the risk of graft failure and leukemic relapse and therefore is not a solution to the GVHD problem. The high degree of variation in the intensity of GVHD observed in different patients suggests that multiple non-MHC genetic factors influence GVHD severity. We hypothesize that, in addition to minor histocompatibility antigen disparities, polymorphisms in genes encoding immunologic effector molecules may be important factors influencing GVHD development. This study aims to explore this hypothesis by identifying non-MHC genes that influence the outcome of BMT in a murine model. In this model, B10.D2 donor leukocytes cause acute GVHD in (C57BL/6xDBA/2)F1 (B6D2F1) recipients, whereas DBA/2 donor leukocytes do not. To date, a locus on chromosome 1 has been identified as influencing the severity of GVHD in this model. Our current study shows that a locus on chromosome 2 acts independently of the chromosome 1 locus to also influence GVHD severity in this model. The region of chromosome 2 implicated in our study contains genes encoding beta2 microglobulin, the minor histocompatibility antigen H-3 and the pro-inflammatory cytokine IL-1. PMID- 10382960 TI - Multicentre European study comparing selection techniques for the isolation of CD34+ cells. AB - Primitive haemopoietic cells are required for studies in both the clinical and research fields and a number of systems have been developed to facilitate isolation of these haemopoietic cell populations. We have analysed the results from several European centres using positive selection of CD34+ cells from haemopoietic tissues (n = 110). Four selection techniques including immunoaffinity columns (Ceprate LC), immunomagnetic beads (Dynabeads, Baxter Isolex 50) and submicroscopic magnetic beads (MACS) were used and the selected CD34+ cells were assessed for purity, yield and enrichment of colony-forming cells (CFC). The mean purities for all samples ranged from 68.4-78.4% for MACS, 33.9-69.9% for Dynabeads, 46.9-66.8% for Ceprate LC and 43.2-65% for Baxter Isolex 50. Yields were variable with all techniques. On average CFC enrichment using the immunoaffinity columns was greater than that observed for the other systems. Some techniques appear to be problematic and may require further expertise to improve the results. Nevertheless, the study demonstrates that highly purified CD34+ cells can be isolated from various haemopoietic sources, though yield and CFC enrichment varies significantly depending on the technique selected. This extends our previous report indicating that not all selection methods generate similar results and that there are differences in the purity, number and colony-forming ability of the cells recovered. PMID- 10382961 TI - Anaplastic squamous cell carcinoma (SCC) in a patient with chronic cutaneous graft-versus-host disease (GVHD). AB - We describe an allogeneic bone marrow (BM) recipient who developed aggressive, metastasizing squamous cell cancer (SCC) of the skin, and discuss possible risk factors in the development of this secondary solid tumor. The patient had been treated with cyclosporine (CsA), methyl-prednisolone and thalidomide for 3 years because of extensive de novo chronic cutaneous GVHD occurring 1 year after BMT. Ten years after BMT a locally invasive and metastasizing SCC occurred on the patient's neck, and diagnosis was confirmed by H&E histopathology and cytokeratin immunohistochemistry. Analysis of genomic DNA did not reveal p53 mutations nor were HPV sequences detectable. Risk factors included conditioning for BMT with total body irradiation (TBI) and cyclophosphamide (Cy), immunosuppressive treatment for GVHD, and extensive exposure to UV radiation before and after BMT. Despite surgery and adjuvant chemotherapy with 5-fluorouracil (5-FU) the patient died 1 year after the diagnosis of SCC. PMID- 10382962 TI - Donor lymphocyte infusion induced molecular remission in relapse of acute myeloid leukaemia after allogeneic bone marrow transplantation. AB - Donor lymphocyte infusion (DLI) has been used successfully to induce remissions in relapse of acute myeloid leukaemia (AML) after bone marrow transplantation (BMT), but molecular eradication of leukaemia has rarely been documented. A patient with AML-M4Eo relapsed after HLA-identical sibling BMT in first complete remission (CR). Cytogenetic and molecular genetic investigations confirmed inv(16) and CBFbeta/MYH11 fusion characteristic of M4Eo. A second remission was obtained with chemotherapy. Full donor chimerism was demonstrated by fluorescence in situ hybridisation. However, molecular evidence of minimal residual disease still persisted, and donor lymphocyte infusion (DLI) was administered. This resulted in molecular eradication, and the patient remained in clinical and molecular remission 16 months from DLI. Our observations showed that, for AML relapse after BMT, molecular leukaemia eradication could be achieved by DLI so that, in cases where genetic markers are available, molecular monitoring should be performed to assess the efficacy of treatment. PMID- 10382963 TI - Successful treatment of refractory acquired pure red cell aplasia (PRCA) by allogeneic bone marrow transplantation. AB - This case describes a 16-year-old woman treated successfully by a bone marrow transplant from her HLA-identical brother for refractory acquired pure red cell aplasia. Conditioning was as for severe aplastic anaemia with cyclophosphamide 4 x 50 mg/kg and antithymocyte globulin. Complete donor type engraftment at 3 months reversed to full autologous reconstitution at 2 years with normal haemopoiesis. The potential implications on pathogenesis of the disease as well as on treatment of autoimmune disorders by stem cell transplantation are discussed. PMID- 10382964 TI - Fulminant adenovirus hepatitis following unrelated bone marrow transplantation: failure of intravenous ribavirin therapy. AB - Fulminant hepatic failure due to adenovirus infection is a rare complication following stem cell transplantation. We report this complication in an unrelated bone marrow transplant recipient 30 weeks post-transplant. Treatment with intravenous ribavirin was started within 36 h of admission, but he succumbed to unusually fulminant hepatic failure. Adenovirus type 2 was isolated from stool surveillance samples and from post-mortem liver samples. Adenovirus DNA was detected by PCR in blood and sputum samples at admission and was identified in post-mortem liver tissue by electron microscopy. Implications of the failure of ribavirin therapy are discussed. PMID- 10382965 TI - Putative mechanism of natural transformation as deduced from genome data. AB - Genetic transformation is widely utilized in molecular biology as a tool for gene cloning in Escherichia coli and for gene mapping in Bacillus subtilis. Several strains of eubacteria can naturally take up exogenous DNA and integrate the DNA into their own genomes. Molecular details of natural transformation, however, remained to be elucidated. The complete genome of a cyanobacterium, Synechocystis sp. PCC6803, has been sequenced. This bacterium has been used to examine functions of a particular gene. The genome is considered to carry information on natural transformable characteristics of Synechocystis. The first step in genetic transformation is the uptake of exogenous DNA. Proteins with non-specific DNA binding features are required, because specificity in the exogenous DNA has not been demonstrated. Such proteins have modules interacting with the phosphate backbone of DNA, including helix-turn-helix modules. Using a consensus pattern of the phosphate-binding helix-turn-helix module, we searched through the genome data of Synechocystis for genes or open reading frame (ORF) products with the pattern in primary structures. We found that an ORF, slr0197, has the pattern in duplicate at the C-terminal region. We also found that the ORF product has a hydrophobic segment at the N-terminal region, which is followed by two internal repeats of the endonuclease domain. Based on these observations, we propose a model for the initial stage of genetic transformation. This is apparently the first report on molecular mechanisms of natural transformation. PMID- 10382966 TI - Complete genome sequence of an aerobic hyper-thermophilic crenarchaeon, Aeropyrum pernix K1. AB - The complete sequence of the genome of an aerobic hyper-thermophilic crenarchaeon, Aeropyrum pernix K1, which optimally grows at 95 degrees C, has been determined by the whole genome shotgun method with some modifications. The entire length of the genome was 1,669,695 bp. The authenticity of the entire sequence was supported by restriction analysis of long PCR products, which were directly amplified from the genomic DNA. As the potential protein-coding regions, a total of 2,694 open reading frames (ORFs) were assigned. By similarity search against public databases, 633 (23.5%) of the ORFs were related to genes with putative function and 523 (19.4%) to the sequences registered but with unknown function. All the genes in the TCA cycle except for that of alpha-ketoglutarate dehydrogenase were included, and instead of the alpha-ketoglutarate dehydrogenase gene, the genes coding for the two subunits of 2-oxoacid:ferredoxin oxidoreductase were identified. The remaining 1,538 ORFs (57.1%) did not show any significant similarity to the sequences in the databases. Sequence comparison among the assigned ORFs suggested that a considerable member of ORFs were generated by sequence duplication. The RNA genes identified were a single 16S-23S rRNA operon, two 5S rRNA genes and 47 tRNA genes including 14 genes with intron structures. All the assigned ORFs and RNA coding regions occupied 89.12% of the whole genome. The data presented in this paper are available on the internet homepage (http://www.mild.nite.go.jp). PMID- 10382967 TI - The gene for an exopolyphosphatase of Pseudomonas aeruginosa. AB - In Pseudomonas aeriginosa, a gene, ppx, that encodes exopolyphosphatase [exopoly(P)ase; EC 3.6.1.11] of 506 amino acids (56,419 Da) was found downstream of the gene for polyphosphate kinase, ppk. Since ppx is located in the opposite direction of the ppk gene, they do not constitute an operon. The predicted amino acid sequence of PPX is 41% identical with Escherichia coli PPX. The gene product of ppx (paPPX) was overproduced in E. coli, and its activity was evaluated. Orthophosphate (Pi) is released from polyphosphate [poly(P)], the average chain lengths of which are 79 and 750, respectively. The amount of Pi released matched the amount of poly(P) lost. Thus ppx encodes an enzyme that has exopoly(P)ase activity. PMID- 10382968 TI - Cloning of cellulose synthase genes from Acetobacter xylinum JCM 7664: implication of a novel set of cellulose synthase genes. AB - Three sets of cellulose synthase genes were cloned from a cellulose-producing bacterium Acetobacter xylinum JCM 7664. One set of genes (bcsAI/bcsBI/bcsCI/bcsDI) were highly conserved with the well-established type I genes in other strains of A. xylinum, while the other two (bcsABII-A, bcsABII-B) were homologous to the known type II (acsAII). Unexpectedly, they were immediately followed by a gene cluster of bcsX/bcsY/bcsCII/ORF569, likely forming an operon. Western blotting demonstrated that the BcsY protein accumulated in cells. Since BcsY showed striking similarities to a number of membrane-bound transacylases, it was hypothesized that the type II cellulose synthase produces acylated cellulose, which might be anchored on the cytoplasmic membrane. An insertion sequence of IS1380-type was found just upstream of the one type II gene (bcsABII-B), suggestive of nonfunctioning. PMID- 10382969 TI - A sequence-ready contig map of the top arm of Arabidopsis thaliana chromosome 3. AB - A fine physical map of the top arm of Arabidopsis thaliana chromosome 3 has been constructed by ordering P1, TAC and BAC clones using the sequences of a variety of DNA markers and end-sequences of clones. The marker sequences used in this study were derived from 58 DNA markers, 93 YAC end-sequences, and 807 end sequences of P1, TAC and BAC clones. The entire top arm of chromosome 3, except for the centromeric and telomeric regions, was covered by a single contig 13.3 Mb long. This fine physical map will facilitate gene isolation by map-based cloning experiments as well as genome sequencing of the top arm of chromosome 3. The map and end-sequence information are available on the web site KAOS (Kazusa Arabidopsis data Opening Site) at [http://www.kazusa.or.jp/arabi/]. PMID- 10382970 TI - Isolation and characterization of rice MADS box gene homologues and their RFLP mapping. AB - Thirty-five MADS box gene homologues were identified through a large-scale cDNA analysis in rice. Based on the nucleotide sequences of the 3'-untranslated region, these clones were classified into 11 independent species. Seven species were found to be new among the rice MADS box gene family, and the other 4 corresponded to the previously reported OsMADS1, OsMADS2, OsMADS4, and OsMADS5. The full nucleotide sequences of the 7 new species were determined. Each clone encoded a deduced protein of 164-267 amino acids. The K-domain of the MADS protein was conserved in all clones though with lower degree in clone S10304. Reverse transcription PCR analysis showed that clones E31254 and E31864 were expressed mainly in panicles. Dendrogram analysis suggested that E31254 and E31864 are close to Arabidopsis AGL9 and AP1, respectively. Restriction fragment length polymorphism (RFLP) linkage mapping revealed that the rice MADS box gene homologues reported here are not clustered but are located throughout the genome. The locus of E31864 on the RFLP map was closely linked to the long sterile lemma gene, g-1. PMID- 10382971 TI - Complete DNA sequence and characterization of a 330-kb VNTR-rich region on chromosome 6q27 that is commonly deleted in ovarian cancer. AB - We report the complete genomic DNA sequence and the characterization of a 330-kb region on chromosome 6q27 that is often deleted in ovarian cancers. Using computer programs to predict exonic sequences, we isolated four novel genes, HGC6.1-4, as well as the known AF-6 gene. None of the deduced products of the novel genes exhibited significant homology to previously known proteins. We also identified ten microsatellites and 12 different VNTR sequences within the target region. HGC6.3 contained a VNTR within a coding exon, each repeat consisting of 42 nucleotides; the predicted 14-amino-acid consensus unit is MTPTVFSSQHTAGG. At least nine different sizes of this VNTR locus were detected among 20 unrelated DNA samples from caucasians. The polymorphic markers and the transcript map documented here may contribute to identification of novel genes or allelic aberrations associated with the development of ovarian cancers. PMID- 10382972 TI - Genomic organization and chromosomal localization of the human cathepsin L2 gene. AB - Cathepsin L2 is a recently described cysteine proteinase with high sequence homology to cathepsin L and other members of the papain superfamily of cysteine proteinases. Its expression is regulated in a tissue-specific manner and is high in thymus, testis and cornea. In the present study, the entire gene sequence, including 5' and 3' flanking region, and chromosomal localization of human cathepsin L2 were determined. The gene spans approximately 6.4 kb and consists of eight exons and seven introns. Genomic organization was similar to human cathepsin L and more than 50% similarity was found between the first introns of cathepsin L and L2, suggesting that they diverged late in evolution. The transcription initiation site, determined by primer extension, was 198 nucleotides from the first ATG. The 5' flanking region lacks a TATA box but has one SP1 site. The gene was mapped to chromosome 9q21-22 by fluorescence in situ hybridization and the distance from cathepsin L was determined to be 15 cM by compiling radiation hybrid mapping results with a genetic map. PMID- 10382973 TI - The so-called chromosomal verotoxin genes are actually carried by defective prophages. PMID- 10382974 TI - Advanced glycation end-products in diabetic nephropathy. AB - Throughout the industrialized (well-fed) world, diabetes mellitus is the most prevalent cause of end-stage renal disease (ESRD). Diabetic nephropathy is as likely to develop in long-duration non-insulin-dependent diabetes (type 2) as in insulin-dependent diabetes mellitus (type 1). Nephropathy in diabetes follows a well outlined course, starting with microalbuminuria through proteinuria, azotaemia and culminating in ESRD. Renal functional decline in diabetic nephropathy is slowed by establishment of euglycaemia and normalization of hypertensive blood pressure. Diabetic ESRD patients, compared with other causes of ESRD, sustain greater mortality and morbidity due to concomitant systemic disorders, especially coronary artery and cerebrovascular disease. A central role for glucose toxicity, especially the adverse impact of accumulated advanced glycosylated end-products (AGEs), appears likely from experimental data generated both in induced diabetic rodents and diabetic individuals. Treatment with aminoguanidine raises the possibility of blocking end-organ damage in diabetes without the necessity for correcting hyperglycaemia. PMID- 10382975 TI - Can we improve the results and increase the number of renal transplants? PMID- 10382976 TI - Organ donation in Spain. PMID- 10382977 TI - Regional integration for widescale transplant coordination: the AIRT model. InterRegional Transplant Association. PMID- 10382979 TI - The pros and cons of renal transplantation in central and southern Italy. PMID- 10382978 TI - Renal transplantation in Sardinia: the local experience and the collaboration with the CCST organization. Consorzio Centro Sud Italia. PMID- 10382980 TI - Evidence-based nephrology. AB - Systematic reviews and meta-analyses are the best approaches available for summarizing the available evidence concerning the efficacy of therapies. Although the renal field has been slow to use these techniques, they are being used increasingly. In March 1997, the Cochrane Renal Group was formed, and this group aims to produce and maintain up to date systematic reviews of the evidence on the effectiveness of therapies used to treat patients with renal diseases. This group is part of the Cochrane Collaboration which is an international structure grouping collaborators together, with the aim of preparing, maintaining and disseminating systematic reviews of the effects of health care in all areas of medicine. PMID- 10382981 TI - Evidence-based medicine: the clinician's perspective. PMID- 10382982 TI - Evidence-based medicine and its horizons: a useful tool for nephrologists? AB - Though the concept of 'evidence-based medicine' (EBM) nowadays has become very popular and even fashionable, its practice is far from being an established reality. There are many reasons why, despite its potential, EBM finds obstacles in expressing its full potential as a tool to better inform health care decisions. Broadly speaking, these obstacles fall into three categories: (i) inadequacy of available information with respect the complexities of health care delivery; (ii) poor quality of clinical research; and (iii) insufficient and inappropriate efforts to promote the uptake of effective interventions in clinical practice. In the first part of the paper, we will discuss: (i) what evidence-based medicine is; (ii) why systematic reviews are the fundamental tool of EBM and what is really special about them; (iii) what are the tools for the practice of EBM; (iv) what its limitations are; and (v) what are the hindrances to its implementation. In the second part, a brief assessment of the state of the art of systematic reviews in nephrology will be presented, with special reference to the activities of the recently launched Cochrane Collaborative Review Group in Renal Diseases. PMID- 10382983 TI - Morphological features of primary focal and segmental glomerulosclerosis. AB - Focal and segmental glomerulosclerosis (FSGS) is one of the most common and non specific patterns of glomerular injury encountered in human renal biopsies. The primary form can be considered when there is a nephrotic syndrome without other causes. The majority of authors agree that podocytes play a role in the development of segmental glomerulosclerosis. The lesion begins with cell hypertrophy, foot process effacement, cell body attenuation, pseudocyst formation, cytoplasmic overload with reabsorption droplets and finally detachment of the glomerular basement membrane (GBM). When the GBM is denuded, it comes into contact with the parietal epithelium and parietal epithelial cells will attach to the GBM, leading to a synechia and, finally, sclerosis. Along this zone, parietal epithelial cells rest on hyalin material. Occlusion and collapse of a group of capillaries is observed, with inclusion of foam cells and hyalin deposits. The origin of primary FSGS has not yet been elucidated. Genetic, racial and developmental factors, macrophages, viral factors and circulating factors are being explored and give encouraging results. PMID- 10382984 TI - Secondary focal and segmental glomerulosclerosis. PMID- 10382985 TI - Clinical picture and outcome of primary focal segmental glomerulosclerosis. AB - Patients with primary focal segmental glomerulosclerosis (FSGS) present with proteinuria (often the nephrotic syndrome), microscopic haematuria, hypertension and renal insufficiency. Overall, this glomerular lesion is seen in approximately 20% of nephrotic adults and children, but is observed much more commonly in the black than the white population (prevalence as high as 80%). Characteristically, nephrotic patients, particularly those with massive proteinuria, have a significantly poorer prognosis than non-nephrotic patients, with 50% progressing to end-stage renal disease (ESRD) over 3-8 years as compared with a 10-year survival of >80%, respectively. In addition, the recurrence rate of this lesion is high in transplanted patients with primary FSGS. When clinical and histological features at presentation have been evaluated by multivariate analysis, the significant positive predictors of progression to ESRD have consistently been the serum creatinine (>1.3 mg/dl), amount of proteinuria and the presence of interstitial fibrosis (> or =20%). The only factor found to be a significant negative predictor of progression to ESRD has been the achievement of a remission in proteinuria. Unfortunately, spontaneous remissions are rare in FSGS, occurring in < or =6% of patients. The factor identified as most associated with achieving a remission in nephrotic patients with primary FSGS has been treatment. PMID- 10382986 TI - Treatment of primary focal segmental glomerulosclerosis. PMID- 10382987 TI - Tropical parasitic nephropathies. PMID- 10382988 TI - On-line monitoring and convective treatment modalities: short-term advantages. AB - BACKGROUND: Despite technological advances in dialysis equipment, the morbidity and quality of life of uraemic patients undergoing regular haemodialytic treatment are still severely affected by acute intradialytic complications possibly related to the treatment itself. Cardiovascular instability still affects >30% of dialytic sessions and, although its pathogenesis is multifactorial, dialysate sodium concentration (and, consequently, intradialytic sodium removal) is one of the main factors affecting intradialytic hypotension. Convective treatment modalities and so-called biocompatible membranes increasingly are recognized as improving acute and particularly chronic dialytic complications because a number of the pathways activated in patients during dialysis with 'bioincompatible' membranes have the potential to produce many side effects. METHODS: The main clinical studies are reviewed to highlight the advantages of on-line monitoring and convective modalities on acute intradialytic symptoms. RESULTS: The conductivity kinetic model has been shown to be a reliable and inexpensive method of matching intradialytic sodium removal and interdialytic load. By applying this model to patients prone to dialysis hypotension, a smaller reduction in intradialytic systolic blood pressure has been observed, without any change in dialysate and reinfusate sodium concentrations or dry body weight. Furthermore, a new model of haemodialysis potassium removal based on a decreasing intradialytic potassium concentration and a constant plasma-dialysate potassium gradient is capable of reducing the arrhythmogenic effect of standard haemodialysis. Despite the proven biological superiority of biocompatible membranes, there is no definitive evidence that membrane biocompatibility and/or flux lead to a decrease in acute intradialytic clinical symptoms. CONCLUSIONS: On line monitoring of intradialytic sodium removal and the potassium gradient is capable of reducing intradialytic hypotension and the arrhythmogenic effect of haemodialysis, and thus having a considerable clinical impact on acute intradialysis complications. As far as the effects of biocompatibility and/or flux on the incidence of acute intradialytic clinical symptoms are concerned, further trials involving a sicker patient population with higher prevalence of intradialytic hypotension are needed in order to achieve statistical power. PMID- 10382989 TI - Ultrafiltration and convective-based dialysis modalities: new trends and applications for renal replacement therapy in ESRD patients. PMID- 10382990 TI - Mechanical strain of glomerular mesangial cells in the pathogenesis of glomerulosclerosis: clinical implications. AB - Due to their elasticity, glomeruli will undergo excessive expansion and repetitive cycles of distension contraction under conditions of impaired glomerular pressure autoregulation and systemic arterial hypertension. These alterations in glomerular volume are associated with mesangial cell stretch which in turn stimulates the synthesis and deposition of ECM with eventual mesangial expansion and glomerulosclerosis. Hyperactivity of growth factors with prosclerotic activity is an important component in the translation of cellular mechanical strain into the abnormal metabolism of ECM components. Although mesangial cell mechanical strain is expected to occur in both remnant glomeruli and in glomeruli of diabetic kidneys, quantitatively different factors will determine the resultant metabolic consequences. In remnant glomeruli, the mechanical stretch is intense, being accounted for largely by the marked glomerular hypertrophy and increased glomerular compliance. In diabetic glomeruli, however, the mechanical stretch is less prominent but its effect on ECM synthesis is markedly aggravated by the presence of hyperglycaemia. There are presently no methods clinically available to diminish the prosclerotic action of growth factors at the glomerular level. In addition, there are no effective means to specifically improve glomerular pressure autoregulation. Therefore, current therapies must be aimed at decreasing systemic arterial pressure, blocking angiotensin II action and reducing glomerular hypertrophy. While there are effective drugs for the treatment of hypertension and for angiotensin II inhibition, protein restriction is the only measure available to diminish glomerular hypertrophy. Finally, in diabetes correction of systemic and glomerular hypertension should be coupled with strict glycaemic control to correct both glomerular autoregulation and increased ECM deposition. PMID- 10382991 TI - Aptamers: novel tools for specific intervention studies. PMID- 10382992 TI - Diuretics in congestive heart failure: new evidence for old problems. PMID- 10382993 TI - How long can dialysis be postponed by low protein diet and ACE inhibitors? PMID- 10382994 TI - Reflex sympathetic dystrophy syndrome in renal transplant patients. A mysterious and misdiagnosed entity. PMID- 10382995 TI - Pathophysiology of ANCA-associated glomerulonephritis. PMID- 10382996 TI - Atrial natriuretic peptide gene delivery attenuates gentamycin-induced nephrotoxicity in rats. AB - BACKGROUND: Atrial natriuretic peptide (ANP) is a cardiac hormone which exerts potent natriuretic and vasorelaxant activities. The aim of this study is to investigate potential protective effects of ANP gene delivery in gentamycin induced nephrotoxicity. METHODS: Adenovirus (Ad.RSV-ANP) carrying the human ANP gene or carrying the LacZ gene (Ad.RSV-LacZ) under the control of the Rous sarcoma virus promoter were delivered intravenously on the first day of gentamycin administration. Sprague Dawley rats were injected subcutaneously with gentamycin daily for 10 days. RESULTS: A single systemic injection of Ad.RSV-ANP at a dose of 1.2x10(10) pfu results in a significant increase in urine excretion, water intake, urinary sodium and potassium excretion. Adenovirus-mediated ANP gene delivery significantly increased renal blood flow, glomerular filtration rates and urine flow as well as attenuated the elevation of blood urea nitrogen levels. Histological evaluations revealed that ANP delivery attenuated gentamycin induced renal tubular damage, cellular necrosis, and lumenal protein casts. The expression of human ANP mRNA was identified in rat kidney, heart, aorta and liver. Immunoreactive human ANP was detected in the heart and kidney of rats injected with Ad.RSV-ANP but not in rats injected with Ad.RSV-LacZ. Cyclic GMP levels in the kidney were significantly increased in rats receiving ANP gene delivery. CONCLUSIONS: This study shows that ANP gene delivery exhibits protection against gentamycin-induced nephrotoxicity and raises the potential to use ANP gene therapy for the treatment of drug-induced renal failure. PMID- 10382997 TI - Overexpression of the human 72 kDa heat shock protein in renal tubular cells confers resistance against oxidative injury and cisplatin toxicity. AB - BACKGROUND: Recent studies have shown that the 72-kDa heat shock protein (HSP72) can be induced in renal tubular cells by a variety of stress conditions, and suggested its cytoprotective function. We have tested this hypothesis directly by transfection studies. METHODS: LLC-PK1 cells (porcine renal tubular epithelial cells) were stably transfected with pBK-CMV or pBK-CMV containing the human HSP72 gene (pBK-CMV-HSP72). These cells were then treated with various concentrations of hydrogen peroxide or cisplatin. The cell viability and lytic cell damage were determined by the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay and lactate dehydrogenase release assay. RESULTS: Immunoblot and immunocytochemical analyses showed the high level expression of HSP72 in LLC-PK1 cells transfected with pBK-CMV-HSP72. In addition, the expression of other major HSPs (HSP90, HSP73, HSP60 and HSP27) was not affected by transfection. LLC-PK1 cells overexpressing HSP72 were significantly more resistant to hydrogen peroxide and cisplatin treatments than control cells. CONCLUSION: These results indicate that overexpressed HSP72 plays a direct role in protecting renal tubular cells against oxidative injury and cisplatin toxicity. PMID- 10382998 TI - In vitro erythrophagocytosis by renal tubular cells and tubular toxicity by haemoglobin and iron. AB - BACKGROUND: Patients with gross haematuria of glomerular origin may develop acute tubular necrosis and reversible renal failure. Erythrocytes within the cytoplasm of proximal tubular epithelial cells (PTECs) can be seen on examination of renal biopsies from these patients. It is possible, therefore, that the tubular damage is a result of cytotoxic breakdown products released during erythrocyte degradation. METHODS: To test this hypothesis, we evaluated (i) by transmission electron microscopy, the capability of a PTEC line to phagocytose and degrade erythrocytes in vitro; and (ii) the effect on the viability of PTCEs in vitro both after erythrophagocytosis and after incubation with haemoglobin, free iron or both. RESULTS: Electron microscopic examination of PTECs exposed to erythrocytes for 96 h showed that 22% of PTECs contained one or more erythrocyte. These were within phagolysosomes and showed varying stages of degradation, with collapse and breakdown of the cell membrane and invasion by cytoplasmic organelles (the so-called haemolytic pathway of erythrocyte degradation). Despite the phagocytosis and degradation of the erythrocytes, no cytotoxicity could be demonstrated under the experimental conditions used. However, the presence of haemoglobin, free iron or both in the culture medium was toxic to the PTECs, resulting in a significant reduction in the number of viable cells present. CONCLUSIONS: PTECs are able to phagocytose and degrade erythrocytes, and haemoglobin and iron are toxic to proximal tubular cells in vitro. PMID- 10382999 TI - L-arginine reduces tubular cell injury in acute post-ischaemic renal failure. AB - BACKGROUND: The pathophysiology of renal ischaemia, resulting in tubular cell injury and leading to acute renal failure (ARF), remains unclear. An ever increasing number of investigations focus on a possible role of nitric oxide (NO) in regulating circulation during ARF. In this context, we investigated the influence of chronic stimulation or inhibition of NO synthesis, or both, on haemodynamic parameters, histology and plasma renin activity (PRA) after ischaemia-reperfusion injury of rat kidneys. METHODS: Experiments were performed on adult, male Wistar rats. Before induction of ARF, a group of animals was treated with a NO synthesis inhibitor (L-NAME) and another group was treated with a precursor of NO synthesis (L-arginine). The animals received those substances for 4 weeks. Control groups received the same amount of tap water for 4 or 8 weeks and were divided into groups with ARF (4 weeks--ARF group and 8 weeks ARF group) and a sham-operated group. Another group of rats was treated first with L NAME and then with L-arginine in their drinking water, for 4 weeks for each of these two substances. All parameters were evaluated 24 h after the induction of ischaemic ARF or the sham operation. RESULTS: Our results show that such long term stimulation of NO release by L-arginine improved renal haemodynamics in the ischaemic form of ARF. Renal blood flow (RBF) increased by 96% in the L-arginine treated rats with ARF compared with the group with ARF alone. Inhibition of NO synthesis worsens renal haemodynamics after ARF. However, this aggravation can be reversed by L-arginine. The rate of water reabsorption was reduced in all groups with ARF, but this reduction was least in the group treated with L-arginine. The rate of Na+ reabsorption was reduced in all groups 24 h after renal ischaemia, but a significant decrease was observed after the inhibition of NO synthesis. Histological examination of the kidney specimens showed that morphological changes were least in the rats treated with L-arginine, when compared with all other groups with ARF. Nevertheless, the lesions were most prominent in the L NAME+ARF group. In this group, the areas of corticomedullar necrosis were more widespread in comparison with other groups, especially the L-arginine group where only swelling of the proximal tubular cells was observed. Treatment with L-NAME was not accompanied by any significant alteration in the plasma concentration of angiotensin I (ANG I), while in the group treated with L-arginine ANG I had a tendency to decrease. CONCLUSIONS: Acute post-ischaemic renal failure may be alleviated by administering the NO substrate (L-arginine). NO acts cytoprotectively on tubular epithelial cells in ischaemia--reperfusion injury of rat kidney. Evidence of this comes from both histopathological findings and increased tubular water and sodium reabsorption. However, inhibition of NO synthesis (provoked by L-NAME) worsens renal haemodynamics and aggravates morphological changes after ARF. These aggravations can, however, be reversed by L-arginine. PMID- 10383000 TI - Enhanced interstitial expression of caldesmon in IgA nephropathy and its suppression by glucocorticoid-heparin therapy. AB - BACKGROUND: With progressive renal disease, structural derangement increasingly encompasses the tubulointerstitial compartment. Tubulointerstitial injury is a critical determinant of renal functional reserve and prognosis in renal disease. Interstitial cells acquiring characteristic of myofibroblasts are an important contributor to interstitial fibrosis. Caldesmon, a calmodulin or actin binding protein, is a molecular marker of differentiation in smooth muscle cells and has recently been shown by us to be a good marker of mesangial cell activation in IgA nephropathy patients. METHODS. We studied whether the expression of caldesmon in interstitium of the kidney was enhanced in the process of glomerular disease and whether it would be a marker of interstitial activation in specific disease states. We performed immunohistochemical staining with anti-caldesmon antibodies in 38 biopsy specimens from IgA nephropathy patients and analysed them quantitatively with a computer-aided manipulator. Interstitial caldesmon expression were compared with histological changes and clinical parameters. RESULTS: Caldesmon expression was enhanced where interstitial cell infiltration and fibrosis were found. Immunoelectron microscopy revealed that caldesmon staining in the renal interstitium was cytoplasmic, and in the processes of myofibroblast-like cells. Caldesmon expression was more prominent in the intense CD68 infiltrated group than in the low positive cells infiltrated group. Patients showing high intensity of interstitial caldesmon expression had significantly higher urinary protein excretion than those showing low intensity of caldesmon expression. Next, 15 patients were treated with glucocorticoid and heparin for 4 8 weeks and re-biopsies were performed. Caldesmon expression was reduced in concomitant with decreased interstitial cell infiltration. Follow-up of these patients (average 24 months) revealed a significant suppression of urinary protein excretion and significant improvement of creatinine clearance. CONCLUSION: These results suggest that the interstitial caldesmon expression is associated with the progression of IgA nephropathy, and glucocorticoid--heparin therapy may reverse the phenotypic change of interstitial cells during the disease process of glomerulonephritis. PMID- 10383001 TI - Anti-DNA antibodies in the urine of lupus nephritis patients. AB - BACKGROUND: It has previously been reported that patients with systemic lupus erythematosus (SLE) and glomerulonephritis do not have anti- (deoxyribonucleic acid) DNA antibodies in their urine. This finding was attributed to specific entrapment of anti-DNA antibodies by the immune complexes in the glomerular capillary walls. METHODS: This phenomenon has been re-investigated as part of a study of the use of desoxyribonuclease 1 (DNase 1) to treat lupus nephritis (LN). For this purpose an ELISA was developed for the detection of anti-DNA antibodies in urine. It was found that such an assay was very susceptible to the presence of DNase in urine which destroys the antigen coating the plates and gives rise to false negative results. For this reason, it is essential that all tests for anti DNA antibodies in the urine are carried out in the presence of EDTA to inhibit the endogenous DNase 1 activity. RESULTS: Using this assay to test the urine from 24 patients with LN and non-selective proteinurea, it was found that they all contained anti-DNA antibodies. The amount of anti-DNA antibodies detected in the urine was compared with that expected by calculations from the anti-DNA antibody titre in the serum and total immunoglobulin levels in serum and in urine. It showed that in 20 patients there was neither specific entrapment nor specific excretion of anti-DNA in urine, only the expected amount of leakage. In only three patients was any appreciable entrapment demonstrated and in only one, any excess excretion. CONCLUSIONS: It is suggested that the failure to detect anti DNA antibodies in the urine in the previous work was due to failure to inhibit the endogenous urinary DNase. It remains to be determined whether the retention of anti-DNA antibodies or excessive secretion is correlated with clinical phases of LN. PMID- 10383002 TI - Urine IgG2/IgG4-ratio indicates the significance of the charge selective properties of the glomerular capillary wall for the macromolecular transport in glomerular diseases. AB - BACKGROUND: Alterations of the charge-selective properties of the glomerular capillary wall are important constituents of the pathogenesis of many glomerular diseases. Thus, differences in the degree of such changes could be of help in understanding the mechanisms governing the transport of macromolecules across the glomerular capillary wall. METHODS: The ratio between urine concentrations of neutral IgG2 and negatively charged IgG4 (IgG2/IgG4-ratio) was measured in 150 proteinuric patients and 21 healthy controls. The patients were subdivided into seven biopsy verified diagnostic groups. RESULTS: The study revealed decreased IgG2/ IgG4-ratio in membranous glomerulonephritis (0.57) compared to healthy controls (2.09) and to all other diagnosis groups; crescentic necrotizing glomerulonephritis (1.28), diffuse proliferative glomerulonephritis (1.10), IgA nephropathy (1.11), mesangial proliferative glomerulonephritis (1.55), minimal change nephropathy (1.00), and nephrosclerosis secondary to hypertension (1.06). Although not statistically significant, there was a tendency towards lower IgG2/IgG4-ratio values in all the studied glomerular diseases compared to healthy controls. CONCLUSIONS: Since IgG is transported entirely through the large pores of the glomerular basement membrane decreased IgG2/IgG4-ratio implies that this pathway is strongly influenced by the charge-selective properties of the glomerular capillary wall. The conclusion that could be drawn from that is that the large pore radius must be discrete, in the order of 80-90 A, and thus not non discriminatory to macromolecules as previously thought. PMID- 10383003 TI - CD4dullCD8bright double-positive T-lymphocytes have a phenotype of granzyme Bpos CD8pos memory T-lymphocytes. AB - BACKGROUND: T-lymphocytes that co-express CD4 and CD8 antigens may be found in small percentages in the peripheral blood of healthy individuals, and have a CD4brightCD8dull phenotype. CD4dullCD8bright T-lymphocytes have been found only in temporal association with some viral infections. METHODS: Four-colour flow cytometric analysis of peripheral blood mononuclear cells from a renal transplant recipient with cytomegalovirus infection was performed. RESULTS: A small but clearly distinguishable subpopulation of CD4dullCD8bright double-positive T lymphocytes was detected, that exhibited phenotypic characteristics of cytotoxic T-lymphocytes and were granzyme B positive. Furthermore, no naive cells appeared to be present within this subpopulation. CONCLUSIONS: CD4dullCD8bright double positive T-lymphocytes are enriched for memory and effector cytotoxic T cells. PMID- 10383004 TI - The relationship between distal tubular proton secretion and dietary potassium depletion: evidence for up-regulation of H+ -ATPase. AB - BACKGROUND: Dietary potassium depletion is associated with elevated plasma bicarbonate concentration and enhanced bicarbonate reabsorption in the distal tubule. The relationship between distal proton secretion and potassium status was investigated by in vivo microperfusion of the superficial distal tubule. METHODS: Experiments were performed on anaesthetized rats that had been maintained on either a low-potassium or control diet for 3-5 weeks prior to experimentation. The distal tubules were perfused at 10 nl/min with either a standard or a barium chloride-containing solution, and the late distal tubular transepithelial potential difference (Vte) and pH of the luminal fluid were recorded using a double-barrelled voltage and ion-sensitive microelectrode. RESULTS: In control rats, the Vte was -40.7+/-2.4 mV and the tubular fluid pH was 6.44+/-0.07; in potassium-depleted animals, the Vte was -15.0+/-1.4 mV and the pH was 6.76+/ 0.03. The pH values in both groups of animals were significantly lower than would be predicted from the Vte and systemic pH for passive H+ distribution, indicating active proton secretion. Moreover, in hypokalaemic rats, this difference from predicted pH was significantly greater than in control animals (control = 0.27+/ 0.06 vs. low-potassium = 0.46+/-0.03; P<0.01), suggesting enhanced active proton secretion. During perfusion with a solution containing BaCl2, the late distal tubule Vte became lumen positive in potassium-depleted rats, contrasting with an increased lumen negativity in potassium-replete controls. The barium-induced lumen-positive potential difference observed in the hypokalaemic rats was abolished by intravenous administration of acetazolamide. CONCLUSION: These data are consistent with enhanced electrogenic proton secretion (H+ -ATPase) during dietary potassium deprivation. PMID- 10383005 TI - Cisplatin therapy in childhood: renal follow up 3 years or more after treatment. Swiss Pediatric Oncology Group. AB - BACKGROUND: In childhood, cisplatin is an essential component of solid tumour therapy such as in neuroblastomas, germ cell tumours, bone tumours, liver tumours and brain tumours. The potential nephotoxicity of cisplatin is widely recognized, but little information is available on permanent sequelae. METHODS: Of the 500 children included in the Swiss Pediatric Oncology Group Late Effect Study, a group of 46 patients (27 males and 19 females) aged 5.7-28 years (median 14 years) surviving the above-mentioned solid tumours entered the present study. The patients were disease-free and off antineoplastic medication for at least 3 years. No recent gastrointestinal or urinary disturbances had occurred, and diets as well as appetites were normal. RESULTS: Blood pressure and plasma or urinary calcium and phosphate were similar in 17 patients treated with cisplatin (dose 142-717, median 400 mg/m2), in 19 patients without cisplatin and in 20 control subjects. A tendency (P<0.02) towards increased plasma creatinine (79 (69-89) micromol/l; median and interquartile range) and low plasma magnesium (0.80 (0.78 0.85) mmol/l) was noted in patients treated with cisplatin as compared with those without cisplatin (68 (58-80) micromol/l; 0.84 (0.79-0.90) mmol/l) and controls (71 (64 80) micromol/l; 0.83 (0.80-0.90) mmol/l). No correlation was noted between the dosage of cisplatin and circulating magnesium or creatinine. CONCLUSIONS: The study demonstrates that the permanent renal disturbances ((i) decreased renal function and (ii) hypomagnesaemia) noted after treatment with cisplatin during infancy or childhood are mild. Furthermore, the study does not demonstrate renal sequelae in patients with the same malignancies who had been treated without cisplatin. PMID- 10383006 TI - The effect of protein restriction on albuminuria in patients with type 2 diabetes mellitus: a randomized trial. AB - BACKGROUND: A randomized trial was conducted to assess whether protein restriction helps to delay the onset of renal disorders in type 2 diabetic patients. METHODS: Included in the trial were 121 type 2 diabetic patients with microalbuminuria or at least detectable albuminuria, or diabetes of duration > or =5 years. The experimental (39 male/19 female) and control group (35 male/28 female) received counselling on protein restriction and the usual dietary advice, respectively. The outcome measure was albuminuria (mg/24 h). Results. After 6 months in experimental and control groups the change in protein intake was 0.05+/-0.21 and +0.03+/-0.19 g/kg (P = 0.02), and in albuminuria -14% and +11% (P = 0.01), respectively. After 12 months, the differences between the experimental and the control group with respect to both protein intake and albuminuria had decreased. At 6 and 12 months, respectively, albuminuria was 28% (P<0.001) and 18% (P = 0.08) lower in the experimental than the control group. The effect in normoalbuminuric patients did not differ from that in microalbuminuric patients. In the experimental group, blood pressure, HbA1c and body weight decreased; in the control group, such decreases were less or absent. Dose-response analysis showed that a 0.10 g/kg change at 6 months in the intake of protein, of animal protein in particular, was related to an 11.1% change in albuminuria (P<0.005). Combining the intakes at 6 and 12 months suggested a percentage change of 9.1%. CONCLUSIONS: Substantial protein restriction in primary care, type 2 diabetic patients with no nephropathy is barely feasible. However, even a small reduction has a substantial and potentially beneficial effect on albuminuria. PMID- 10383007 TI - Preservation of renal function by percutaneous transluminal angioplasty in ischaemic renal disease. AB - BACKGROUND: The purpose of this study was to evaluate the effects of percutaneous transluminal renal angioplasty (PTRA) on preservation of renal function in patients with bilateral renal artery stenoses or stenosis of the artery of one functioning kidney. METHODS: A total of 227 PTRAs of 223 stenoses in 135 patients were performed from 1982 to 1993 in a single centre and retrospectively reviewed. The number of PTRAs per patient was 1.7, range 1-6. Angiographical follow-up was performed in 77%, 120+/-82 days after the first PTRA and 273+/-345 days after the last PTRA. Follow-up of serum creatinine and blood pressure was performed in 85% after 414+/-558 days. Long-term follow-up was performed for dialysis, surgical revascularization, renal transplantation and death, mean follow-up 8.8 years, range 5.5-14.8. RESULTS: The immediate technical success was 90%, and another 5% were improved. The primary patency rate per patient was 43% and the secondary patency rate 64%. Improved renal function was achieved in 23% of the patients, stabilized in 56% and failed in 21%. Stabilized or improved function was higher when baseline serum creatinine was < or =250 micromol/l (85%) than >250 micromol/l (60%). Three of 99 (3%) patients with creatinine < or =250 micromol/l started dialysis during follow-up (41 days, 7.4 and 8 years), as did 13 of 36 (36%) patients with creatinine >250 micromol/l. Blood pressure and the number of antihypertensive drugs decreased in patients with creatinine < or =250 micromol/l, but was unchanged in those with creatinine >250 micromol/l. The 5 year survival rates were 84, 66 and 17% for patients with creatinine <125 micromol/l, 125-250 micromol/l and >250 micromol/l, respectively. Twelve patients (9%) experienced complications, including two deaths. CONCLUSIONS: Our study shows that PTRA improved or preserved the renal function in most patients with normal to moderately impaired renal function. Close follow-up and possibly re intervention are necessary to obtain satisfactory clinical and angiographical result. PMID- 10383008 TI - Hepatic HDL receptor, SR-B1 and Apo A-I expression in chronic renal failure. AB - BACKGROUND: Chronic renal failure (CRF) is associated with hypertriglyceridaemia and depressed plasma high-density lipoprotein (HDL)-cholesterol and apolipoprotein A-I (Apo A-I) concentrations. Uraemic hypertriglyceridaemia is due, in part, to lipoprotein lipase and hepatic lipase deficiencies, which are causally linked to excess parathormone (PTH). This study was designed to test the hypothesis that depressed plasma concentration and abnormal composition of HDL in CRF may be due to dysregulation of hepatic expression of Apo A-I and/or the newly discovered HDL receptor. METHODS: Hepatic Apo A-I and HDL receptor mRNA abundance (Northern blot), and HDL receptor protein mass (Western blot) were determined in CRF rats (5/6 nephrectomy), parathyroidectomized CRF rats (CRF-PTx) and sham operated controls. RESULTS: The CRF group exhibited normal hepatic HDL receptor mRNA and HDL receptor protein abundance coupled with reduced hepatic Apo A-I mRNA. Hepatic Apo A-I mRNA, HDL receptor mRNA and protein abundance were not affected by PTx. CONCLUSIONS: CRF results in the down-regulation of hepatic Apo A I gene expression, which accounts for the known reduction in plasma Apo A-I concentration. However, CRF does not affect HDL receptor mRNA or protein expression in this model. Neither Apo A-I nor HDL receptor expression were modified by PTx in CRF rats. PMID- 10383009 TI - Clinical, radiological and serum amyloid P component scintigraphic features of beta2-microglobulin amyloidosis associated with continuous ambulatory peritoneal dialysis. AB - BACKGROUND: Beta2-Microglobulin (beta2M) amyloidosis occurs in patients with end stage renal failure (ESRF) who undergo long-term continuous ambulatory peritoneal dialysis (CAPD), but its prevalence in patients treated exclusively by CAPD is unknown. In addition, its features may differ from those of haemodialysis associated beta2M amyloidosis because CAPD is more biocompatible. METHODS: We performed serum amyloid P component (SAP) scintigraphy, a specific technique for imaging amyloid deposits, in 13 consecutive patients with ESRF who had been dialysed for >5 years, at least 80% of the time by CAPD. Clinical and radiological features of beta2M amyloidosis were sought and compared with the results of SAP scintigraphy. RESULTS: SAP scans showed articular amyloid deposits in seven patients, all of whom had evidence of carpal tunnel syndrome and four of whom had arthralgia characteristic of dialysis amyloidosis. Typical radiographic bone cysts were present in only one case who had been dialysed for >17 years. The remaining six patients had no clinical, radiological or scintigraphic evidence of beta2M amyloidosis. CONCLUSIONS: The prevalence of beta2M amyloidosis in this study was comparable with that in reported haemodialysis populations. Many of the amyloid deposits demonstrated by SAP scintigraphy were not associated with symptoms, but larger and longer term studies are required to determine whether CAPD favourably influences their clinical expression. PMID- 10383010 TI - Low levels of dehydroascorbic acid in uraemic serum and the partial correction of dehydroascorbic acid deficiency by haemodialysis. AB - BACKGROUND: Vitamin C is currently considered a potent water-soluble antioxidant and it appeared reasonable to study the metabolic changes of vitamin C in uraemia and during haemodialysis. METHODS: We measured the levels of ascorbic, dehydroascorbic and diketogulonic acids in sera of uraemic patients before and during haemodialysis, using the 2,4-dinitrophenylhydrazine method. RESULTS: The results indicate that the levels of ascorbic and dehydroascorbic acids in uraemic sera are low in comparison with controls, but the levels of diketogulonic acid are higher than in healthy persons. The comparison of ratios between levels of these substances in uraemic sera and in normal sera indicate that the oxidation of ascorbic acid to dehydroascorbic in uraemia proceeds much slower than in controls. We propose that uraemic patients are characterized by pronounced deficiency of dehydroascorbic acid. A marked decrease of ascorbic and diketogulonic acid plasma levels, and a pronounced increase of dehydroascorbic acid levels during haemodialysis was observed. Ratios between plasma levels of ascorbic and dehydroascorbic acids following haemodialysis are the same as in healthy persons. CONCLUSION: Uraemic patients are characterized by marked deficiency of dehydroascorbic acid and this deficiency can be partially corrected by haemodialysis. PMID- 10383011 TI - Calcium ketoglutarate versus calcium acetate for treatment of hyperphosphataemia in patients on maintenance haemodialysis: a cross-over study. AB - Since dietary restrictions and phosphorus removal by haemodialysis (HD) are not sufficient to control serum phosphate (s-phosphate) levels in dialysis patients the use of oral phosphate binders is mandatory. Calcium ketoglutarate (CaKE) is an analogue of glutamic acid exerting phosphate binding properties. Therefore we compared this substance to calcium acetate (CaAC) in a 24-weeks open cross-over trial in 28 maintenance HD patients. Medications and HD prescriptions were kept unchanged during the trial. Following 2 weeks of withdrawal of phosphate binders, patients were randomly assigned to one of the calcium salts for 12 weeks; after a second withdrawal of 2 weeks, all patients were shifted to the other treatment for another 12 weeks. All patients received equimolar doses of CaKE and CaAC with respect to the amount of prescribed elemental calcium. Treatment with CaAC and CaKE significantly reduced s-phosphate levels after 4 weeks (CaAC 1.95+/-0.6 vs. 2.4+/-0.53 mmol/l, P = 0.004; CaKE 1.95+/-0.4 vs. 2.47+/-0.63 mmol/l, P = 0.0001) reaching a virtually stable plateau over the remaining observation time without significant differences between the groups. The incidence of hypercalcaemia defined as a serum calcium level > or =2.8 mmol/l was significantly higher in CaAC than in CaKE treated patients (n = 8 vs. n = 1, P = 0.03). There were no significant differences in serum intact parathyroid hormone (PTH) bicarbonate, albumin or calcitriol levels between the groups after 12 weeks treatment. We conclude that CaKE is as effective as CaAC for treatment of hyperphosphataemia in chronic HD patients and may be particularly helpful in patients who are prone to develop hypercalcaemia. PMID- 10383012 TI - Prognostic value of heart rate variability during long-term follow-up in chronic haemodialysis patients with end-stage renal disease. AB - BACKGROUND: Mortality is high in chronic haemodialysis patients with cardiovascular disease, and many of them die suddenly. Reduced heart rate variability (HRV) is an increased risk for death in various populations, but its prognostic value in haemodialysis patients remains uninvestigated. METHODS: We analysed the associations between 24-h HRV measures and long-term mortality through a prospective follow-up of 31 chronic haemodialysis patients who underwent diagnostic coronary angiography. RESULTS: Of the 31 patients, at baseline, seven had a previous myocardial infarction, five had a history of congestive heart failure and 14 had significant (> or =75%) coronary stenosis (four had multi-vessel stenosis). During follow-up for 60+/-5 months, 14 patients died, 11 of them suddenly. A left ventricular ejection fraction of <0.45, multi vessel coronary stenosis, ventricular tachycardia on 24-h ECG and decreased/abnormal 24-h HRV (triangular index <22 and abnormal Poincare plot) carried a univariate risk of all-cause death, while the risk of sudden death was only correlated with decreased HRV (standard deviation of normal-normal R-R interval <50 ms, triangular index <22 and ultra-low frequency power <8.7 ln(ms2)). Multivariate analysis revealed that a triangular index <22 was the best predictor of increased risk for both all-cause and sudden death (hazards ratio (95% CI); 8.1 (1.3-48.6) and 12.6 (1.3-126.4), respectively) and that the association was independent of cardiac function, macrovascular diseases, ventricular arrhythmias and cardiovascular risk factors. The 5-year mortality when the triangular index was > or =22 or <22 was 33 or 88% for patients with coronary artery disease and 0 or 50% for those without. CONCLUSIONS: These results indicate that HRV has an independent prognostic value in chronic haemodialysis patients and identifies an increased risk for all-cause and sudden death. PMID- 10383013 TI - Prognostic value of cardiac troponin T and I elevations in renal disease patients without acute coronary syndromes: a 9-month outcome analysis. AB - BACKGROUND: Moderate elevations of cardiac troponin (Tn) T, up to levels presumably diagnostic for minor myocardial damage, are suspected to be false positive in nearly 0.3 of end-stage renal disease (ESRD) patients undergoing haemodialysis (HD). It is not clear whether cardiac TnI is superior to TnT in those patients, if differences between ESRD and pre-ESRD occur, and what the prognostic meaning of these troponin elevations might be. SUBJECTS AND METHODS: We examined 40 chronic renaldisease patients [56.4 SD 13.9 years; 22 male, 18 female) without evidence of an acute coronary syndrome (ACS) for at least 28 days prior to the investigation. Cardiac status was determined by history, physical examination, ECG and echocardiography. Patients were divided into subgroups with HD (n = 20) and without HD (n = 20). Patients without HD had a mean creatinine clearance (CC) of 13.45 ml/min. Tn were measured by immunoassay techniques. TnT was compared to two different TnI tests (TnID, TnIB), CK/CKMB activity and myoglobin (MYO) concentrations. In all patients, a 9-month follow-up for acute myocardial infarction, re-hospitalization, and death was completed. RESULTS: None of the troponins significantly predicted patient outcome. Tn did not correlate with CC (r<0.6). Applying the lowest reported threshold values for all tests in the HD group, 0.3 patients were positive for TnT, 0.55 patients were positive for TnID, and 0.15 for TnIB. In the group without HD, 0.2 patients were positive for TnT and TnID and 0.1 for TnIB. CONCLUSIONS: Moderate elevations of cardiac troponins are common in clinically stable patients with renal disease and are neither diagnostic for an acute coronary syndrome nor predictive of outcome. It is concluded that increased troponins in asymptomatic renal patients are of questionable value for risk stratification, most probably due to unspecific elevations. PMID- 10383014 TI - Ambulatory nocturnal oximetry and sleep questionnaire-based findings in 38 patients with end-stage renal disease. AB - BACKGROUND: Patients with end-stage renal diseases (ESRD) have an increased risk of sleep-disordered breathing. With regard to this disorder, controversy persists about prevalence, cost-effective assessment and socio-economical relevance. METHODS: Therefore, we performed, for the first time, overnight ambulatory oximetry in combination with a sleep questionnaire in 38 unselected patients with ESRD and 37 healthy controls. An oxygen desaturation index (ODI) >15, defined as >15 falls in oxygen saturation of > or =4% per h, was observed more frequently in ESRD patients than in healthy controls (47 vs. 3%, P<0.001). RESULTS: In general, the results derived from the assessment of the Epworth Sleepiness Scale (ESS) as well as those from the visual analogue scale (VAS) did not reflect the ODI values of the respective patient population. Interestingly, 88% of ESRD patients with the questionnaire finding 'excessively loud snoring' had an ODI of >15 as compared with 13% without this complaint (P<0.05). Furthermore, 77% of ESRD patients with a systolic blood pressure >140 mm Hg and a body mass index (BMI) >25, had an ODI of >15. The percentage of ESRD patients with a professional activity was higher in the absence of sleep-disordered breathing (63 vs. 21%, P<0.05). CONCLUSION: 'Excessively loud snoring' and a BMI >25 combined with hypertension are risk factors for sleep-disordered breathing in ESRD patients. Nocturnal oxygen desaturations are assessed efficiently by ambulatory oximetry and correlate with relevant biological and socio-economical parameters in ESRD patients. PMID- 10383015 TI - Health-related quality of life in dialysis patients. A report from an Italian study using the SF-36 Health Survey. DIA-QOL Group. AB - BACKGROUND: Interest in measuring health-related quality of life (HRQoL) has increased together with an awareness that such humanistic outcomes require valid and reliable measures. In the last decade short, simple and multidimensional generic and disease-specific questionnaires have been developed. Among the several generic questionnaires available, the Short Form 36 Items Health Survey (SF-36) was translated and validated in several languages, and applied to different settings and diseases. METHODS: Within the framework of a larger, prospective, multicentre study (DIA-QOL project) the SF-36 was administered to 304 patients to test its characteristics in terms of patient acceptability, and psychometric and clinical validity. Standard psychometric techniques were used to evaluate its validity in terms of convergence, divergence and internal consistency reliability (Cronbach's alpha). Correlations between clinical variables and HRQoL scores were performed to test the questionnaire's capability to capture differences across patients groups. RESULTS: Overall, the findings show that, in this sample, the SF-36's performance was very good. Acceptability was satisfactory, with a response rate higher than 80%. All the questionnaire scales met the psychometric standards suggested in terms of grouping and scaling assumptions. The internal reliability coefficients actually replicate the satisfactory findings reported previously for the original SF-36. In terms of the ability of the questionnaire scales to discriminate between groups expected to differ in a given health concept in relation to clinical variables, the results were also good. On average, females reported lower scores, the impact of ageing was more evident for physical scales. Diabetic patients score significantly worse on the physical function scale and patients with mental health problems score significantly lower on the mental health scale. No significant association was found with the index KtV, haemoglobin levels, body mass index, parathyroid hormone and type of dialysis. A strong association was indeed found between SF-36 scales measuring physical health concepts and the serum albumin level. This association held after adjusting for the confounding effect of age. Comparison of the health profile of the present sample with others from the US and UK and from a representative sample of the Italian general population highlights the potential of such questionnaire in dialysis setting. CONCLUSIONS: The SF-36 questionnaire is easy to use in Italian dialysis patients and SF-36 scores are related to important clinical aspects. This approach can help in caring for dialysis patients and can be useful in outcome assessment programmes. PMID- 10383016 TI - Pregnancy in women receiving renal dialysis or transplantation in Japan: a nationwide survey. AB - BACKGROUND: Since a report on the first successful pregnancy of a woman on long term haemodialysis in Japan in 1977, there has been a growing number of case reports on successful pregnancy in patients on dialysis. We undertook a nationwide survey on pregnancy in women on renal replacement therapy in 1996. METHODS: A preliminary questionaire was sent to 2504 dialysis units and 143 renal transplant units in Japan. For each reported pregnancy, a more detailed questionaire was sent to collect nephrological, obstetric and neonatal information. RESULTS: There were 172 pregnancies (0.44%) reported in 38889 women on dialysis, with 90 successful pregnancies (0.23%), and 194 pregnancies reported in 852 female renal transplant recipients. Detailed pregnancy information was collected from 74 women on dialysis and 194 renal transplant recipients. Of the 74 pregnancies in the women on dialysis, 36 (48.6%) resulted in surviving infants, nine (12.2%) in neonatal death, nine (12.2%) spontaneous abortions and 14 (18.9% elective abortions were reported. The outcome of six pregnancies (8.1%) was unknown. Of 194 pregnancies in renal transplant recipients, 159 (82.0%) resulted in surviving infants, two (1.4%) in neonatal death and 28 (14.4%) in spontaneous or elective abortion. In five cases the pregnancy outcome was not reported. No congenital anomalies were reported, except two infants with mental retardation and one with epilepsy. CONCLUSION: The current survey revealed that the rate of successful pregnancy in women on dialysis has improved. More than half of the pregnancies resulted in infant survival. But, premature birth is a major problem for the children of women on dialysis and there is a higher rate of neonatal death. There are significant differences in gestational age, birth weight, frequency and severity of prematurity and rates of neonatal death between pregnancies of women undergoing dialysis and those who are renal transplant recipients. PMID- 10383017 TI - Hydroxyethyl starch does not impair immediate renal function in kidney transplant recipients: a retrospective, multicentre analysis. AB - BACKGROUND: The effect of hydroxyethyl starch (HES) on early allograft function was examined retrospectively in a cohort of 119 renal transplantations realized by local organ exchange between four cooperating centres. METHODS: After exclusions, 109 transplant procedures were subdivided in three groups according to donor colloid loading: (i) HS-group (Haes steril 6%, mean volume (SD): 979 (946) ml, n = 20); (ii) PS-group [Plasmasteril, mean volume (SD): 769 (411) ml, n = 16]; and (iii) control group (gelatin albumin, n=73). RESULTS: Delayed graft function (DGF), defined as the need for dialysis during the first post-transplant week, occurred in 3/20 (15%) cases in the HS-group, in 5/16 (31%) cases in the PS group and in 14/73 (19%) cases in the control group (P = 0.450). Uni- and multivariate analysis revealed older donor age (P = 0.001) and kidney preservation with histidine-tryptophan-ketoglutarate (HTK) (P = 0.001) as the only factors associated with a higher incidence of DGF. CONCLUSIONS: Renal function as measured by daily serum creatinine concentration and 24 h urinary output up to 14 days post-transplantation in the HS-group was comparable with that of controls. The higher serum creatinine observed during the first seven post-transplant days in the PS-group could be related to higher donor age and haemodynamic instability, and recipient male preponderance, rather than to HES itself. PMID- 10383018 TI - Ureteral complications in renal transplantation with more than one donor ureter. AB - BACKGROUND: The purpose of this study was to evaluate the ureteral complications of renal transplant recipients with more than one donor ureter METHODS: Between 1967 and 1997, 19 patients (median age 34 years, range 6-62 years) received renal transplants from donors with more than one ureter. There were 18 donor organs with two ureters, and one patient underwent en bloc renal transplantation with four donor ureters. In nine patients, the ureters were implanted separately at the bladder dome according to the extravesical technique of Witzel, Sampson, Lich and Rohl. In 10 patients, we performed a modification of this extravesical technique according to Nghiem with a side-to-side anastomosis of the ureters before completing the ureteroneocystostomy. RESULTS: After a median follow-up of 55 months (range 2-218 months), no graft loss due to ureteral complications was noted. One patient died due to myocardial infarction, seven patients returned to dialysis without ureteral complications. There were two patients (one patient after side-to-side ureteral anastomosis, one patient with separate implantation of the two ureters) with ureteral obstruction of one donor ureter. Both patients underwent open surgical revision with temporarily placement of internal ureteral stents. CONCLUSIONS: The presence of multiple ureters from donor kidneys is associated with a higher complication rate in our patient population compared with donor kidneys with one ureter. There was no difference in the long-term outcome between the two implantation techniques used. PMID- 10383020 TI - Comparison of icodextrin and glucose solutions for the daytime dwell in automated peritoneal dialysis. AB - BACKGROUND: The sustained ultrafiltration achieved by icodextrin is more suited for the daytime dwell in automated peritoneal dialysis (APD) than glucose solutions. METHODS: Seventeen patients receiving APD underwent assessment using three different solutions for the daytime dwell: 2.27% glucose, 3.86% glucose and 7.5% icodextrin. Patients were then observed on icodextrin for a 6 month period. RESULTS: Daytime ultrafiltration was greater for 3.86% glucose (median 0.10, IQR 0.01 to 0.321) P<0.01 and icodextrin (median 0.26, IQR 0.14 to 0.361) P<0.001 than 2.27% glucose (median -0.19, IQR -0.54 to -0.081), with 3.86% glucose and icodextrin not being significantly different. Positive ultrafiltration occurred in 3/17 patients with 2.27% glucose, 13/17 patients with 3.86% glucose and 16/17 patients with icodextrin (chi2 P<0.0001). The difference in ultrafiltration of icodextrin and 3.86% glucose correlated with the 4 h dialysate/plasma creatinine ratio in a PET test (r = 0.51, P<0.05). Daytime Kt/V urea was greater for 3.86% glucose (median 0.27, IQR 0.20 to 0.48 per week, P<0.01) and icodextrin (median 0.31, IQR 0.27 to 0.49 per week, P<0.0001) than for 2.27% glucose (median 0.22, IQR 0.15 to 0.38 per week), with the difference between 3.86% glucose and icodextrin not reaching statistical significance (P = 0.06). Daytime creatinine clearance was greater for 3.86% glucose (median 10.2, IQR 6.9 to 13.61/week/1.73 m2, P<0.02) and icodextrin (median 12.1, IQR 9.3 to 15.71/week/1.73 m2, P<0.005) than for 2.27% glucose (median 8.8, IQR 4.9 to 11.91/week/1.73 m2). Daytime creatinine clearance was greater for icodextrin than for 3.86% glucose (P<0.005). The effects of icodextrin were sustained for the 6 month observation period. CONCLUSIONS: Icodextrin produced enhanced ultrafiltration and clearances compared with 2.27% glucose, without the exposure of the peritoneum to hypertonic glucose solutions. PMID- 10383019 TI - Altered flow properties of blood and increased plasma fibrinogen in cyclosporin treated renal allograft recipients. AB - BACKGROUND: Abnormalities in blood rheology may be factors contributing to cardiovascular complications and the progression of renal failure in kidney allograft recipients. The haemorheological variables haematocrit, fibrinogen, whole blood viscosity, plasma viscosity, erythrocyte aggregation tendency and fluidity were measured in 27 cyclosporin A (CyA)-treated patients who had received a renal graft at least 6 months previously. Their creatinine clearance was in the range of 12-92 ml/min/1.73 m2 (mean 55+/-19). The values were compared with those obtained from a control group comprising 20 healthy subjects matched according to age, sex and smoking habits. RESULTS: The haematocrit, plasma fibrinogen, whole blood viscosity, plasma viscosity, erythrocyte aggregation tendency, body mass index (BMI), mean arterial pressure (MAP) and serum triglycerides were increased in the transplanted patients, and the serum high density lipoprotein (HDL)-cholesterol and erythrocyte fluidity decreased. The haemorheological variables were used as dependent variables in a stepwise regression analysis with age, MAP, BMI, urinary albumin excretion rate, blood CyA concentration, creatinine clearance, and serum triglycerides, cholesterol and HDL cholesterol as independent variables. Plasma fibrinogen was positively correlated with BMI and blood CyA. The whole blood viscosity was positively correlated with blood CyA and negatively with serum HDL-cholesterol. Only serum triglycerides remained correlated with erythrocyte aggregation tendency. CONCLUSIONS: All variables with a known impact on blood viscosity were altered in the present group of renal transplant recipients. Inappropriate regulation of erythrocyte formation, overweight, the use of CyA, high triglycerides and low HDL-cholesterol levels may be factors contributing to this. The importance of impaired flow properties of blood for the development of cardiovascular diseases and transplant glomerulosclerosis needs to be examined. PMID- 10383021 TI - Prevalence of hypertension in patients on peritoneal dialysis: results of an Italian multicentre study. AB - BACKGROUND: The tenet that peritoneal dialysis is capable of either normalizing or improving blood pressure control in uraemic patients is based on outdated or monocentric experiences. Therefore, we assessed the prevalence of hypertension and the efficacy of antihypertensive therapy in a large, multicentric cohort of patients on peritoneal dialysis. METHODS: Twenty seven out of the 50 centres belonging to the Italian Co-operative Peritoneal Dialysis Study Group took part in the study. The main patient selection criteria were: peritoneal dialysis therapy for at least 3 months and no peritonitis or changes in dialysis technique for at least 1 month. Clinical blood pressure was measured according to WHO/ISH guidelines. Ambulatory blood pressure monitoring was carried out using a SpaceLabs 90207 recorder. Hypertension was defined according to WHO/ISH criteria and staged according to the criteria of the Joint National Committee on Detection, Evaluation and Treatment of High Blood Pressure (JNC), 5th Report. Ambulatory blood pressure monitoring recordings were used to evaluate white-coat hypertension, blood pressure load and the dipping phenomenon. RESULTS: Five hundred and four subjects were evaluated. Hypertension was prevalent in 88.1% of the population, and 362 out of 444 hypertensive patients were on antihypertensive therapy. JNC staging revealed that 188 patients had moderate to severe hypertension. Blood pressure load was pathological in 77.3% of the patients receiving antihypertensive treatment. White-coat hypertension was identified in 9.1% of the hypertensive patients not on antihypertensive therapy, and 53.1% of the patients were non-dippers. CONCLUSIONS: The study demonstrates that hypertension is a dramatic, unsolved problem in uraemic patients treated with peritoneal dialysis, and casts doubts on the effectiveness of our current peritoneal dialysis strategies and pharmacological management of hypertension. PMID- 10383022 TI - Accumulation of advanced glycation end products in the peritoneal vasculature of continuous ambulatory peritoneal dialysis patients with low ultra-filtration. AB - BACKGROUND: Ultra-filtration failure is a serious complication of long-term continuous ambulatory peritoneal dialysis (CAPD). This complication is related to histological changes of the peritoneum, i.e. severe interstitial fibrosis and microvascular sclerosis. Although their pathogenesis has not been elucidated yet, advanced glycation end products (AGEs) have been shown to accumulate in the peritoneal tissue of CAPD patients. METHODS: Peritoneal biopsy specimens from 14 CAPD patients with low ultra-filtration (n = 9) and high ultra-filtration (n = 5) capacity were immunohistochemically investigated using a monoclonal antibody against AGEs (6D12). The severity of peritoneal fibrosis, microvascular sclerosis and intensity of AGE accumulation were semi-quantitatively evaluated. Peritoneal ultra-filtration capacity was evaluated by calculating daily ultrafiltration volume per body weight (UFV/BW) and D/D0 (glucose) of the peritoneal equilibration test. RESULTS: In all patients with low ultra-filtration, AGE accumulated in the peritoneal fibrous tissue and microvascular walls. Remarkably, AGE accumulated more intensely in hyalinized fibrosis of small venular media. Extent of AGE accumulation in peritoneal interstitium and vascular walls correlated with the progression of interstitial fibrosis (rho = 0.727, P = 0.0088) and vascular sclerosis (rho = 0.915, P = 0.001). UFV/BW was inversely correlated to interstitial fibrosis (rho = -0.660, P = 0.0174), microvascular sclerosis (rho = -0.671, P = 0.0155) and microvascular AGE accumulation (rho = 0.678, P = 0.0145). CONCLUSIONS: In CAPD patients, AGE formation in the peritoneum correlates with the development of severe interstitial fibrosis and microvascular sclerosis, which is associated clinically with impaired peritoneal ultra-filtration. PMID- 10383023 TI - Delayed complications following Tenckhoff catheter removal. AB - BACKGROUND: Tenckhoff catheter placement is well established to facilitate continuous ambulatory peritoneal dialysis (CAPD) in the treatment of end-stage renal failure. Complications of these catheters while in situ are well documented. However, little information is available concerning post-removal complications. Many centres, including our own remove these catheters by traction resulting in retained cuffs, rather than by formal dissection. We have evaluated the outcome of such removal over a 2-year period. METHODS: Sixty-two patients underwent Tenckhoff catheter removal by traction over a 2-year period at our unit. Patients were evaluated retrospectively using case notes and operation records. RESULTS: The catheters were sited for a mean of 23 months and were most commonly removed because of persistent peritonitis (48.4%). Sixty-one per cent of all patients had experienced at least one episode of CAPD peritonitis while the catheter was in situ, but this did not correlate with those who developed local sepsis. Fifteen patients (24.2%) subsequently developed local infective complications after a mean of 5.7 months (range 1-17 months). The subcutaneous cuff was involved in all cases and the peritoneal cuff was involved in six cases. Thirty patients were identified as being immunosuppressed, but this was not a risk factor in the development of retained cuff infections. CONCLUSIONS: There is a significant risk of local sepsis with retained cuffs resulting from removal by traction and our data suggests that these catheters should be removed by dissection and excision of both cuffs. PMID- 10383024 TI - New technique of parathyroidectomy to prevent parathyromatosis and hypoparathyroidism. AB - A 54-year-old woman with end-stage renal disease and on haemodialysis for 4 years developed severe secondary hyperparathyroidism and was operated upon. The two upper and the largest lower parathyroid glands were resected. The right lower gland was dissected from the lower pole of the thyroid and, by gently pulling upwards, the lateral walls were dissected using electrocautery. The lower aspect of the gland maintained the blood supply through small mediastinal and thymic vessels of the neopedicle, which allowed its mobilization to a more superficial plane. Because of the large size of the gland, the part opposite to the neopedicle was resected and the cutting surface was sealed with fibrin adhesive. Pre-thyroidal muscles were reapproximated and the remnant of the parathyroid gland was pulled out through a small hole in the inferior part of the midline and sutured with fine silk to the muscle. The gland was therefore placed in a subcutaneous position in the lowest part of the operative field just above the sternal border. The postoperative course was uneventful and, 8 months after surgery, the patient maintains a normal parathyroid function. PMID- 10383025 TI - Hypernatraemia and polyuria due to high-dose colchicine in a suicidal patient. PMID- 10383026 TI - Successful treatment of HCV-associated cryoglobulinaemic glomerulonephritis with a combination of interferon-alpha and ribavirin. PMID- 10383027 TI - Oncogenic osteomalacia in a patient with a fibrocystic nodule of the breast. PMID- 10383028 TI - The value of computed tomography and magnetic resonance imaging to diagnose rhabdomyolysis in acute renal failure. PMID- 10383029 TI - Cure of apparent end-stage renal disease in a patient with dissecting aneurysm of the aorta using a percutaneous interventional approach. PMID- 10383030 TI - Fibromuscular dysplasia of the allograft renal artery with early post-operative dissection: successful treatment with intravascular stent insertion. PMID- 10383031 TI - Angiomyolipoma in a transplanted kidney. PMID- 10383032 TI - Nocardial endophthalmitis leading to blindness in a renal transplant recipient. PMID- 10383033 TI - Acute renal failure in the course of HIV infection: a single-institution retrospective study of ninety-two patients and sixty renal biopsies. AB - BACKGROUND: Acute renal failure syndromes are frequently encountered in patients with human immunodeficiency virus (HIV) infection. Most reported cases of acute renal failure are related to acute tubular necrosis, but many other causes of renal failure have been described in these patients. METHODS: The present work is a single-institution retrospective study of 92 HIV-infected patients with acute or rapidly progressing renal failure. In 60 cases, a renal biopsy was performed. For each patient we analysed clinical and pathological data, as well as the short term prognosis. RESULTS: Ten different causes of acute or rapidly progressing renal failure were documented: (i) haemolytic uraemic syndrome (32 patients); (ii) acute tubular necrosis either of ischaemic-toxic origin (18 patients) or due to rhabdomyolysis (six patients); (iii) obstructive renal failure which was either extrinsic (two patients), drug-induced (13 patients) or secondary to paraprotein precipitation (one patient); (iv) HIV-associated nephropathy (14 patients); (v) acute interstitial nephritis (two patients); (vi) various glomerulonephritis (four patients). In most cases, renal failure was severe (the mean creatinine clearance at entry was 12 ml/min). Most patients had a significant improvement in renal function with only symptomatic treatment. Eighteen per cent of the patients died within 2 months of the diagnosis of renal failure. Renal biopsy seems important for the diagnosis but also for the prognosis, at least in the cases of haemolytic-uraemic syndrome, HIV-associated nephropathy and drug-induced micro-obstructive renal failure. CONCLUSION: Vascular and glomerular diseases are frequent causes of acute or rapidly progressing renal failure in HIV-infected patients. Renal biopsy appears to be safe and useful for the diagnosis and the prognosis of the renal failure. High mortality rate is only observed in patients with ischaemic/toxic causes of acute renal failure. PMID- 10383034 TI - Paroxysmal nocturnal haemoglobinuria. MRI of renal cortical haemosiderosis in two patients, including one renal transplant. PMID- 10383035 TI - Hydrothorax complicating continuous ambulatory peritoneal dialysis: successful management with talc pleurodesis under thoracoscopy. PMID- 10383036 TI - A pain in the arm. PMID- 10383037 TI - Glomerular lipids in non-hereditary forms of glomerulopathy/glomerulonephritis. PMID- 10383038 TI - Acid-base abnormalities in a patient with hepatic cirrhosis. PMID- 10383039 TI - Is diagnosis and treatment of renovascular stenosis cost efficient? PMID- 10383040 TI - Increased plasma GDNF levels in patients with chronic renal diseases. PMID- 10383041 TI - Continuous veno-venous haemofiltration versus continuous veno-venous haemodialysis in severe lithium self-poisoning: a toxicokinetics study in an intensive care unit. PMID- 10383042 TI - Decrease in serum pentosidine levels of ESRD patients during polysulfone haemodialysis. PMID- 10383043 TI - Neisseria subflava biovar perflava peritonitis in a continuous cyclic peritoneal dialysis patient. PMID- 10383044 TI - Conversion between cyclosporin and tacrolimus--30-fold dose prediction. PMID- 10383045 TI - Spontaneous iliac artery dissection in a kidney transplantation treated with and endovascular stent. PMID- 10383046 TI - Characterization of glycine transport in cultured Muller glial cells from the retina. AB - Rapid termination of the synaptic action of glutamate (Glu) and glycine (Gly) is achieved by uptake into the presynaptic terminal and glial cells. In the vertebrate CNS, Gly acts both as an inhibitory neurotransmitter and as a Glu modulator or coagonist at postsynaptic N-methyl-D-aspartate (NMDA) receptors. We have previously described NMDA receptors in Muller cells of chick retina coupled to the phosphoinositide cascade, the entry of calcium, and the activation of protein kinase C (PKC; Lopez-Colome et al. Glia 9:127-135, 1993). A colocalization of Gly transporters and NMDA receptors has been reported in brain tissue (Smith et al. Neuron 8:927-936, 1992); since the concentration of Gly could participate in the modulation of Glu excitatory transmission in the vertical pathways of the retina, transport of Gly in monolayer cultures of Muller cells was studied. Gly transport was found pH-sensitive with an optimum at pH 7.4. Kinetic analysis of the saturation curve for Gly within a concentration range of 0.01-2 mM, revealed two components of transport: a low-affinity system with Km = 1.7 mM, Vmax = 30 nmol/10 min/mg protein, and a high-affinity one with a Km = 27 microM, Vmax = 3 nmol/10 min/mg protein. Both systems were Na+ dependent; the high-affinity system proved also dependent on external Cl- and was inhibited by sarcosine, characteristic of GLYT1 transporters. The inhibition of low-affinity uptake by 2-(methylamino)isobutyric acid (MeAIB) and 2 aminoisobutyric acid (AIB) suggests the presence of transport system A in Muller cells. The process is energy-requiring, since Gly transport was decreased by metabolic inhibitors. Data obtained are in keeping with a modulatory role for Muller glia on excitatory transmission in the retina. PMID- 10383047 TI - Endothelial nitric oxide synthetase (eNOS) in astrocytes: another source of nitric oxide in neocortex. AB - The distribution of the endothelial form of nitric oxide synthetase (eNOS) was examined in the visual cortex of three species of primate and in the rat using immunocytochemistry. Labeled cells were found in both the gray and white matter. These cells were stellate in appearance and labeled cell processes were seen contacting blood vessels or the pia, suggesting that, by morphological criteria, the cells were astrocytes. All eNOS positive cells were double labeled with an antibody against S100beta. Although all cells were double labeled in the white matter, in the gray matter, some S100beta positive cells did not contain detectable levels of eNOS. eNOS positive astrocytic processes appeared to form prominent and distinctive structures next to neurons, especially in cortical layer IIIC. We postulate that these eNOS-positive structures form astrocytic perisynaptic sheaths on neuronal somas in the cortex. If this is true, then nitric oxide can influence neuronal transmission directly at axosomatic synapses in the cortex. In addition, the presence of eNOS in astrocytes and in their processes that contact blood vessels suggests that the link between local cortical activity and changes in cerebral blood flow could be mediated by astrocytic release of nitric oxide. PMID- 10383048 TI - GDNF family members and their receptors: expression and functions in two oligodendroglial cell lines representing distinct stages of oligodendroglial development. AB - Glial cell line-derived neurotrophic factor (GDNF), neurturin (NTN), and persephin (PSP) constitute a subfamily of transforming growth factor-betas (TGF betas) with prominent roles in the regulation of neuron survival and differentiation. Although numerous members of the TGF-beta superfamily are important regulators of glial cell functions in health and disease, it is unknown whether any member of the GDNF subfamily may have functions in normal or pathological glial cell performances. To begin to address this issue, we have studied expression and putative functions of GDNF, NTN, PSP, and their receptors in two cell lines representing models for oligodendrocyte progenitor cells (OLI neu) and immature oligodendrocytes (OLN-93), respectively. RT-PCR analysis revealed expression of all three growth factor mRNAs in OLI-neu and OLN-93 cells. Expression was weak in OLI-neu cells, while both NTN and PSP mRNAs were strongly expressed in OLN-93 cells. Furthermore, OLI-neu and OLN-93 cells expressed transcripts encoding the GDNF receptors Ret and GFRalpha-1. The two splice variants for GFRalpha-2 were exclusively synthesized in OLI-neu cells. Similarly, primary O-2A progenitor cells and enriched mature oligodendrocytes expressed Ret, GFRalpha-1 and GFRalpha-2 mRNAs. Both GDNF and NTN stimulated DNA synthesis monitored by BrdU incorporation of OLI-neu cells in a dose-dependent fashion. Co administration of TGF-beta significantly reduced this effect. Similarly, PDGF co applied with GDNF or NTN down-regulated proliferation in OLI-neu cells. In contrast, OLN-93 cells did not respond to GDNF or NTN with increased incorporation of BrdU. Expression of GDNF, NTN, and their receptors and distinct effects in two model cell lines of oligodendrocyte development suggest that functions of members of the GDNF family and their receptors may not be restricted to neurons and may be implicated in oligodendrocyte development. PMID- 10383049 TI - Sodium-bicarbonate cotransport in retinal astrocytes and Muller cells of the rat. AB - Sodium-bicarbonate cotransport in retinal glial cells was studied in the everted eyecup preparation of the rat. Intracellular pH was monitored with the indicator dye BCPCF and fluorescence confocal microscopy. Raising the K+ concentration from 3 to 12 mM in HCO3- -buffered perfusate evoked an intracellular alkalinization in both astrocytes and Muller cells. The alkalinization developed more rapidly and was larger in astrocytes. The K+ -induced alkalinization was HCO3- -dependent; it was reduced by 33% in astrocytes and 71% in Muller cells when HCO3- was removed from the perfusate. The alkalinization was effectively blocked by addition of 0.5 mM 4,4"-diisothiocyanato-stilbene-2,2'-disulfonic acid (DIDS). Removal of Na+ from the perfusate evoked a rapid acidification in both types of glial cells. The results indicate that astrocytes and Muller cells in situ in the rat retina possess an electrogenic Na+/HCO3- cotransporter. PMID- 10383050 TI - Behaviour of DRG sensory neurites at the intact and injured adult rat dorsal root entry zone: postnatal neurites become paralysed, whilst injury improves the growth of embryonic neurites. AB - The dorsal root entry zone is a PNS-CNS junction between Schwann cells and astrocytes, defining the site where dorsal root ganglia (DRG) axons enter the adult mammalian spinal cord. Following dorsal root injury (rhizotomy), DRG axons regenerate within the PNS environment of the root but stop at the DREZ and fail to re-enter the spinal cord. We have used an in vitro model to compare how neurites growing from embryonic (E13) and postnatal (P0 and adult) DRG neurons behave at the uninjured and rhizotomized adult rat DREZ. We find that both freshly dissected and conditioned-lesioned postnatal DRG neurons seldom grow neurites across cryosections of the uninjured or rhizotomized DREZ. However, embryonic DRG neurons more readily grow neurites across cryosections of the uninjured and 7-day post-lesion (dpl) DREZ and are dramatically better able to cross the 21 dpl DREZ. This enhanced growth was abolished by co-incubation with a function-blocking antiserum to beta1-integrin receptors, whilst immunoreactivity for some beta1-integrin ligands (tenascin-C and fibronectin) increased at the DREZ by 21 dpl, suggesting that beta1-integrin ligands may stimulate the growth of embryonic neurites across the 21 dpl DREZ. Fluorescence time-lapse video microscopy was used to record the behaviour of dye-labelled postnatal DRG neurites as they encounter the uninjured adult DREZ in vitro. Neurites rarely turned around at the DREZ, but instead became paralysed. Of a variety of chemical modifications to uninjured DREZ cryosections, only treatment with methanol, chloroform, or the protease inhibitor D-phe-pro-arg chloromethylketone hydrochloride (PPACK, 100 microM) caused any increase in the proportion of postnatal neurites which crossed the DREZ. PMID- 10383051 TI - Endothelin-induced protein tyrosine phosphorylation of cultured astrocytes: its relationship to cytoskeletal actin organization. AB - Endothelins (ETs) promote cytoskeletal actin reorganization of cultured astrocytes (Koyama and Baba, Neuroscience 61:1007-1016, 1994; Koyama and Baba, Glia 16:342-350, 1996). In this study, we examined the signal transduction involved in that activity of ETs. Immunoblot analysis with an anti phosphotyrosine antibody showed that ET-3 (1 nM) increased tyrosine phosphorylation of 120 Kda and 70 Kda astrocytic proteins. The tyrosine phosphorylations of both proteins reached a maximum at 1 nM ET-3. In morphological examinations, ET-3 (1 nM) induced stress fibers, an organized F actin structure, and focal adhesions in 0.5 mM dibutyryl cAMP (DBcAMP)-treated astrocytes within 30 min. Immunochemical staining of phosphotyrosine revealed that the newly formed focal adhesions possessed phosphotyrosine immunoreactivity. Phorbol 12-myristate 13 acetate (PMA, 100 nM), bradykinin (1 microM), angiotensin II (100 nM), and A23187 (5 microM) did not induce astrocytic stress fibers and had no obvious effects on tyrosine phosphorylation of 120 Kda and 70 Kda proteins. Tyrosine phosphorylation of astrocytic 120 Kda and 70 Kda proteins was stimulated by 1 mM sodium orthovanadate (VO4(3-)), a protein tyrosine phosphatase inhibitor. VO4(3-) promoted reorganization of stress fibers and focal adhesions in DBcAMP-treated astrocytes. Neither chelation of intra- and extracellular Ca2+ nor pre-treatment with pertussis toxin (PTX) affected the ET-induced tyrosine phosphorylation and stress fiber formation in cultured astrocytes. These results suggest a relationship between cytoskeletal actin reorganization and the tyrosine phosphorylation of astrocytic proteins by ETs. PMID- 10383052 TI - De novo expression of lipocortin-1 in reactive microglia and astrocytes in kainic acid lesioned rat cerebellum. AB - An understanding of the role of reactive glia in the neurodegenerative/regenerative process requires a knowledge of the molecules synthesised by these cells following trauma. We investigated the cellular localisation of lipocortin-1 (LC-1), a putative neuroprotective agent, in cryostat sections of normal and kainic acid lesioned rat cerebellum. In the normal cerebellum lipocortin-1 immunoreactivity was detected in Purkinje cell bodies and molecular layer interneurons. Following kainic acid (1 microg) induced lesions, it was rapidly upregulated in activated microglia, from which it appeared to be secreted. At later time points it was detected in activated astrocytes. LC-1 protein levels were quantified by a sensitive and specific ELISA. Compared to control cerebellum, LC-1 levels were dramatically elevated following lesion, peaking at 3 days: 760% of basal (unlesioned) levels. In situ hybridisation studies revealed a marked upregulation of LC-1 mRNA at 1 and 3 days following the lesion, indicating the transient de novo synthesis of this protein, consistent with a localisation to microglia. In vitro studies, on cultured astrocytes and microglia, demonstrated high levels of intracellular LC-1 in both cell types. LC-1 was detected in microglial but not astrocytic, conditioned media, confirming the in vivo observations that activated microglia may secrete LC-1. Our data show that at early time points following excitotoxic lesion to the cerebellum, it is activated microglia that synthesise and possibly secrete this protein, suggesting an important role of this cell type in immunosuppression and neuroprotection following damage to the central nervous system. PMID- 10383053 TI - Signal transduction pathways induced by GM-CSF in microglia: significance in the control of proliferation. AB - Communication between cells of the central nervous system (CNS) and of the immune system is accomplished by a network of cytokines and growth factors. Certain cytokines and growth factors cause activation of microglia, contributing to inflammatory states in the CNS. Granulocyte-macrophage colony-stimulating factor (GM-CSF) has numerous effects on microglia, ranging from induction of proliferation to changes in morphology. GM-CSF is also a growth factor for cells of the myeloid lineage, and the signal tranduction induced by GM-CSF in these cells has been extensively studied. Most notably, the importance of the Jak/STAT and MAP kinase pathways in mitogenesis has been shown in many different systems. We show here that primary microglia and a microglia cell line, BV-2, have a Jak/STAT expression pattern and GM-CSF inducibility similar to that of monocytes and macrophages. Primary microglia and BV-2 cells expressed identical Jak/STATs: Jakl, Jak2, Jak3, Tyk2, STAT1alpha/beta, STAT3, STAT5A, STAT5B, and STAT6. In addition, GM-CSF induced Jak2, STAT5A, and STAT5B in BV-2 cells, as it does in monocytes and macrophages. Immunocytochemical analysis showed that STAT5 translocates to the nucleus following GM-CSF stimulation of microglia. We also found the MAP kinases, ERK1 and ERK2, to be phosphorylated in microglia and BV-2 cells following induction by GM-CSF. Jak2, STAT5A, STAT5B, and ERKs are known to be important in controlling cellular proliferation. Drugs that block these pathways may become tools to control inflammation in the CNS by limiting microglial proliferation. PMID- 10383054 TI - Prosaposin: a myelinotrophic protein that promotes expression of myelin constituents and is secreted after nerve injury. AB - Recently, we demonstrated that prosaposin and prosaptides (peptides encompassing the neurotrophic sequence in prosaposin) prevent cell death and increase extracellular regulated kinase (ERK) phosphorylation and sulfatide content in primary Schwann cells or oligodendrocytes (Hiraiwa et al., 1997a). Here, we examine the effect of prosaptide on other myelin constituents, on Schwann cell morphology and proliferation, and characterize the time course of expression of prosaposin protein after sciatic nerve injury. After 24 h of treatment with 10 nM TX14(A), a 14-mer prosaptide, the specific activity of UDP-galactose:ceramide galactosyltransferase (GalT) in primary Schwann cells was increased by 150% over controls. Under the same conditions, the maximum content of sulfatide increased 3 fold over controls after 48 h of treatment. Northern blot analysis, probed with oligonucleotide sequences from the GalT and P0 cDNAs, revealed that the mRNA levels of GalT and P0 protein were elevated about 30 and 200%, respectively, over controls after 24 h of treatment with TX14(A). Treatment of primary Schwann cells with TX14(A) also induced a morphological change at 10 nM; the peptide-treated cells had a bipolar (spindle-shaped) appearance after 48 h of treatment, compared to control cells which were irregular and multipolar. TX14(A) did not induce cell proliferation, indicating that TX14(A), unlike IGF-I, is not mitogenic. After sciatic nerve transection, Western blot analysis demonstrated the presence of intact prosaposin in tubular fluid in a silicon chamber into which the proximal and distal nerve stumps were sutured. The concentration of prosaposin in the fluid was maximum after 9 days post-surgery and returned to normal after 28 days post-surgery. In uninjured and injured nerve, prosaposin immunolocalized to the smooth muscle of epineurial and endoneurial vessels. These findings indicated that sciatic nerve secreted prosaposin after injury and that prosaposin is a naturally occurring injury-repair protein which acts to prevent degeneration and to promote regeneration of peripheral nerves. PMID- 10383055 TI - Substratum of pleiotrophin (HB-GAM) stimulates rat CG-4 line oligodendrocytes to adopt a bipolar morphology and disperse: primary O-2A progenitor glial cells disperse similarly on pleiotrophin. AB - Pleiotrophin (HB-GAM), an extracellular matrix-associated protein with a high content of basic amino acid residues, is expressed in the central nervous system during late pre- and early post-natal development and promotes neurite outgrowth in vitro. Here, we show that, on a substratum of pleiotrophin formed from a 5 or 10 microg/ml solution, undifferentiated rat CG-4 line oligodendrocytes adopt a bipolar morphology and disperse over the substratum, as we have previously shown with poly-L-lysine (Rumsby et al. Neurosci. Res. Commun. 23:101-109, 1998). On pleiotrophin substrata formed from coating solutions of 1 microg/ml and below, CG 4 line cells form aggregates and do not disperse, as is also the case with poly-L lysine. The same dispersing effect is observed with rat primary 0-2A progenitor glial cells on pleiotrophin substrata from solutions of 5 and 10 microg/ml: 0-2A cells aggregate together on pleiotrophin substrata formed from lower concentrations and do not disperse. A pleiotrophin substratum enhances proliferation of CG-4 line oligodendrocytes and primary 0-2A progenitor glial cells. The results show that pleiotrophin provides a substratum that can influence progenitor oligodendrocyte morphology, aid cell dispersion, and perhaps also enhance progenitor oligodendrocyte cell growth. PMID- 10383056 TI - Will DOTS do it? A reappraisal of tuberculosis control in countries with high rates of HIV infection. AB - In 1993 the WHO declared tuberculosis a global emergency, and subsequently introduced the DOTS strategy, a technical and management package based on earlier work of the IUATLD and international experience with directly observed therapy. Despite successful implementation of most of the elements of this strategy in several African countries and settings, tuberculosis case rates continue to escalate where the prevalence of HIV infection is high. We explore possible reasons for the failure to control tuberculosis even in the context of tuberculosis programmes that have been considered models for others to emulate. In many African countries half or more of tuberculosis patients are now HIV infected; in such settings, the overall epidemiology of tuberculosis is disproportionately affected by what happens in the HIV-infected subpopulation of the community. Persons with HIV infection are at increased risk of rapid progression following primary infection or re-infection, and also from reactivation of latent infection with Mycobacterium tuberculosis. More intensive strategies need to be targeted to the HIV-infected to interrupt on-going transmission (active and passive case detection; prevention of nosocomial transmission) and reactivation (preventive therapy). The high burden of other HIV related disease in patients with tuberculosis, such as other bacterial infections, toxoplasmosis and other manifestations of AIDS, require that tuberculosis programmes integrate their activities better with those of HIV/AIDS programmes, including those for provision of HIV/AIDS care. Enhanced epidemiological surveillance is required to follow tuberculosis trends in the HIV positive and negative sub-populations of communities, which may respond differently to control efforts. Strategies for tuberculosis control programmes in countries of high and low HIV prevalence cannot be the same, but must take into account the epidemiology of HIV infection. HIV/AIDS in Africa poses severe challenges of purpose and identity to tuberculosis control programmes, which have not adapted to the altered realities of the HIV/AIDS era. DOTS alone is unlikely to control tuberculosis in sub-Saharan Africa; one major achievement of DOTS when implemented, however, has been its apparent ability to limit the development and spread of drug resistance. PMID- 10383057 TI - Prevalence of primary and acquired resistance of Mycobacterium tuberculosis to antituberculosis drugs in Benin after 12 years of short-course chemotherapy. AB - SETTING: Benin National Tuberculosis Programme, West Africa. OBJECTIVE: To measure the prevalence of primary and acquired resistance of Mycobacterium tuberculosis to the antituberculosis drugs isoniazid, rifampicin, ethambutol and streptomycin in Benin from 1994-1995, after 12 years of short-course chemotherapy regimens. METHODS: Prospective study by cluster sampling according to the methodology recommended by the International Union Against Tuberculosis and Lung Disease (IUATLD) and the World Health Organization (WHO). RESULTS: The survey of primary resistance included 333 strains, of which 28 (8.4%) were drug-resistant, one to both rifampicin and isoniazid (multidrug-resistant). For acquired resistance, out of 57 strains tested 26 (45.6%) were resistant, six of which (11%) were multidrug-resistant. CONCLUSION: Despite the considerable increase in the number of tuberculosis cases observed in recent years (52% between 1987 and 1995), direct observation of patients taking their antituberculosis drugs during the intensive phase of treatment has limited the development of drug resistance in Benin. PMID- 10383058 TI - The 1997 Nationwide Tuberculosis Prevalence Survey in the Philippines. AB - SETTING: The Philippines is a developing country where tuberculosis (TB) remains a significant public health problem. OBJECTIVE: To determine the prevalence of TB as a basis for setting the targets of the National Tuberculosis Control Program. STUDY POPULATION AND METHODS: A multi-stage cluster survey of a random sample of 21960 subjects from 36 clusters nationwide was undertaken from 2 April to 31 July 1997. BCG scar verification and tuberculin testing was performed for subjects aged 2 months and over, and chest radiography screening was done on subjects 10 years and older. Sputum samples were collected from individuals who were initially assessed to have abnormal chest radiographs to determine the prevalence of bacillary tuberculosis. Acid-fast smear by modified Kinyoun's technique and culture on Lowenstein Jensen were done to demonstrate Mycobacterium tuberculosis. RESULTS: The prevalence of active pulmonary TB was 42/1000 population. The prevalence of culture-positive and smear-positive cases was 8.1 and 3.1/1000, respectively. The prevalence was similar in urban and rural areas. CONCLUSION: Morbidity from TB remains high. Allowing for methodological differences from the survey in 1981-1983, the prevalence of active pulmonary TB was unchanged. There was only a minimal decrease, of 37% for smear-positive cases and 25% for culture positive cases, in the 14-year interval. PMID- 10383059 TI - Prevalence of sputum-positive pulmonary tuberculosis in tribal and non-tribal populations of the Ashti and Karanja tahsils in Wardha district, Maharashtra State, India. AB - SETTING: Ashti and Karanja tahsils, Wardha district, Maharashtra State, Central India. OBJECTIVE: To find and compare the prevalence of bacillary positive pulmonary tuberculosis amongst the different tribes and in the non-tribal population. DESIGN: Prevalence study of pulmonary tuberculosis by house-to-house survey of symptoms among tribal (n = 20596) and non-tribal (n = 93 670) populations aged 5 years and over, between September 1989 and November 1990. RESULTS: The prevalence of smear and/or culture-positive tuberculosis/100000 population was 133 in the tribal and 144 in the non-tribal population. The difference in prevalence of symptomatic individuals and sputum-positive cases among the tribal and the non-tribal populations was statistically significant only in the symptomatic individuals/100000 (P = 0.01). The prevalence of cases in both groups was higher in males than females; however this difference was significant only in the tribal group (P = 0.05). Only two of the 46 tribes encountered, the Mana and Pawara tribes, showed a high prevalence, of 730 and 612/100000, respectively. The three other tribes with positive cases (the Gond group) had prevalences comparable to that of the nontribal population. CONCLUSION: The prevalence of tuberculosis in tribal people was comparable to that of the non-tribal population. PMID- 10383060 TI - The importance of quality control of sputum smear microscopy: the effect of reading errors on treatment decisions and outcomes. AB - SETTING: Ho Chi Minh City, Vietnam. OBJECTIVE: To evaluate the impact of slide reading errors at peripheral level on case-finding and treatment decisions. DESIGN: Over a 6-month period in 1997, information on date, type of slide, results of other slides from the patient, and treatment status was collected for all slides from district TB centers detected as having reading errors during smear microscopy quality control re-readings. RESULTS: Reading errors were detected in 117 slides: 115 (98.3%) were incorrectly read as negative, and 75 (65.2%) of these errors occurred in case-finding slides. In the 75 falsely negative case-finding slides, re-reading resulted in initiation of treatment in 38 patients (50.7%). The remaining 37 (49.3%) had only one positive slide and were told to return for follow-up after 6 months; the two (5.4%) who did return were both diagnosed with active TB. Detection of errors in the 40 false-negative follow-up slides resulted in treatment changes in four patients (10%). CONCLUSIONS: Quality control plays a critical role in helping to ensure the timely diagnosis and treatment of new TB cases and appropriate management of patients currently on treatment. The usefulness of quality control could be enhanced by focusing greater efforts on case-finding slides initially read as negative. PMID- 10383061 TI - The epidemiology of tuberculosis diagnosed after death in San Francisco, 1986 1995. AB - SETTING: San Francisco, California. OBJECTIVES: To identify the characteristics of persons in whom tuberculosis was diagnosed after death, and determine whether secondary cases of tuberculosis resulted from them. DESIGN: Retrospective review of all cases of tuberculosis reported in San Francisco from 1986 through 1995, combined with a prospective evaluation of the molecular epidemiology of tuberculosis. RESULTS: Four per cent of the reported 3102 tuberculosis cases were diagnosed after death. The rate of tuberculosis cases diagnosed after death was 1.63 per 100000 population. Age 43 years or older, male sex, white race, and birth in the United States were characteristics independently associated with a diagnosis of tuberculosis after death. During 1993-1995, injecting drug use was also independently associated with a diagnosis of tuberculosis after death (odds ratio 9.24, 95% confidence interval 1.77-39.38). Cases of tuberculosis diagnosed after death do not appear to be significant sources of undetected tuberculosis transmission causing new secondary tuberculosis cases in the community. CONCLUSIONS: Health care providers in San Francisco, and probably other urban areas, should maintain a high index of suspicion for tuberculosis in ageing, white, US-born males, and injecting drug users. PMID- 10383062 TI - Cutaneous tuberculosis: a twenty-year prospective study. AB - SETTING: A tertiary care hospital in northern India. OBJECTIVE: To study the patterns of clinical presentation of cutaneous tuberculosis, to correlate them with Mantoux reactivity and BCG vaccination status, and to suggest a clinical classification based on these factors. DESIGN: Analysis of the records of patients with cutaneous tuberculosis who attended the hospital between 1975 and 1995. RESULTS: A total of 0.1% of dermatology patients had cutaneous tuberculosis. Lupus vulgaris was the commonest form, seen in 154 (55%) of these patients, followed by scrofuloderma in 75 (26.8%), tuberculosis verrucosa cutis in 17 (6%), tuberculous gumma(s) in 15 (5.4%) and tuberculids in 19 (6.8%). No correlation was found between Mantoux reactivity and the extent of disease (localised disease 63.6%, disseminated disease 67.9%). The presence of regional lymphadenopathy was an indication of dissemination of the disease (localised disease 34.7%, disseminated disease 71.7%). Dissemination of the disease was observed in the whole of the spectrum of cutaneous tuberculosis (22.1%), but was seen more often in the presence of gumma and scrofuloderma. There were more unvaccinated individuals in the group with disseminated disease (80.3%) than in those with localised disease (65.5%). CONCLUSIONS: Lupus vulgaris was the most common clinical presentation, followed by scrofuloderma, tuberculids, tuberculosis verrucosa cutis and tuberculous gumma. Some patients presented more than one clinical form of the disease. Classification of cutaneous tuberculosis needs to be modified to include smear-positive and smear-negative scrofuloderma apart from the inclusion of disseminated disease. The presence of regional lymphadenopathy serves as a clinical indicator of disseminated disease. Patients with disseminated disease were less likely to have been BCG-vaccinated than those with localised disease. PMID- 10383063 TI - Mycobacterium kansasii and M. scrofulaceum isolates from HIV-negative South African gold miners: incidence, clinical significance and radiology. AB - SETTING: A South African gold mining hospital. OBJECTIVE: To investigate the clinical significance of non-tuberculous mycobacteria (NTM) isolates, and estimate NTM disease incidence in human immunodeficiency virus (HIV) negative miners. DESIGN: Retrospective case series describing clinical and radiological features associated with NTM sputum isolates from HIV-negative miners between January 1993 and July 1996, and a comparison group with Mycobacterium tuberculosis infection. RESULTS: Of miners with NTM isolates, 90% had been HIV tested and 81% were HIV-negative. M. kansasii and M. scrofulaceum accounted for 202 (68%) and 41 (14%) isolates respectively. More than 80% of miners with M. kansasii or M. scrofulaceum were smear positive, and new cavitation was present in 78% and 74% respectively. Treatment failure occurred in 3% of M. kansasii and 12% of M. scrofulaceum patients. A normal pre-morbid radiograph was significantly less common in NTM than M. tuberculosis patients (odds ratio 0.26 and 0.10 for M. kansasii and M. scrofulaceum, respectively). NTM disease incidence, defined as NTM isolate plus new cavitation, was estimated at 66 and 12 per 100000 person years for M. kansasii and M. scrofulaceum, respectively. CONCLUSIONS: M. kansasii and M. scrofulaceum disease are common in HIV-negative South African gold miners. Most isolates are associated with new cavitation against a background of silicosis or old TB scarring. PMID- 10383064 TI - Evaluation of a commercial ligase chain reaction assay for the diagnosis of pulmonary and extra-pulmonary tuberculosis. AB - SETTING: Egas Moniz Hospital, Lisbon, Portugal. OBJECTIVE: To evaluate the Ligase Chain Reaction (LCx) Mycobacterium tuberculosis Assay for the direct detection of M. tuberculosis complex in respiratory specimens after smear observation, and its suitability for non-respiratory clinical specimens. DESIGN: Analysis of 156 specimens collected from 123 patients with pulmonary tuberculosis and/or extrapulmonary involvement. RESULTS: Among 93 pulmonary secretions and 63 extra pulmonary samples and after resolution of discrepancies based on clinical and laboratory findings, two pulmonary samples from a patient with a diagnosis of sarcoidosis, four samples of cerebrospinal and one of seminal fluid were considered as false positives. Two tissue biopsy samples, one pericardial effusion and one pulmonary secretion from patients strongly suspected of having tuberculosis were considered as false negatives for the assay, without inhibition of amplification. All specimens yielding M. avium on culture were LCx negative. CONCLUSION: The LCx Mycobacterium tuberculosis Assay was found to be useful for the rapid identification of M. tuberculosis complex in all types of specimens. It revealed a high specificity both in pulmonary and extrapulmonary products, and a sensitivity of 97% for the pulmonary secretions and of 75% for the extra pulmonary specimens, independently of the bacilloscopy results. PMID- 10383065 TI - Chemotherapy of tuberculosis in mice using single implants of isoniazid and pyrazinamide. AB - OBJECTIVE: To establish the chemotherapeutic value of a depot drug preparation of isoniazid and pyrazinamide against experimental tuberculosis. DESIGN: To see whether sustained levels of pyrazinamide are available for prolonged periods after a single subcutaneous administration of a biodegradable polylactic-glycolic acid (PLGA) polymer containing the drug, studies were done to ascertain whether a single administration of isoniazid and pyrazinamide in separate PLGA polymers could offer chemotherapeutic protection against a heavy intravenous challenge of susceptible mice with a virulent strain of Mycobacterium tuberculosis similar to that rendered by daily administration of the two drugs for 8 weeks. RESULTS: Even with three times the daily dose of pyrazinamide contained in the single PLGA polymer implant, no abnormally high (burst) levels of the drug were evident after administration, but sustained levels of the drug were seen up to 54 days. The chemotherapeutic activity of the single PLGA polymer implants was similar to that obtained with standard oral treatment with the two drugs given daily for the entire 8 weeks, as judged by mortality and colony forming unit (CFU) counts of tubercle bacilli from lungs and spleen. CONCLUSION: Treatment with single implants of the PLGA polymer containing anti-mycobacterial drugs offers a strong possibility of circumventing the compliance problem. PMID- 10383067 TI - Street talk: knowledge and attitudes about tuberculosis and tuberculosis control among homeless adults. AB - OBJECTIVES: To measure knowledge and perceived susceptibility to tuberculosis among homeless adults in San Francisco and attitudes toward control measures used to improve adherence to treatment for tuberculosis. DESIGN: A cross-sectional survey via interview of homeless shelter residents was done at five shelters. RESULTS: Of 292 persons interviewed, 21.6% reported a positive skin test, and 57.1% of the positives had received preventive therapy. Over 60% had misconceptions about transmission, in particular confusion with transmission of the human immunodeficiency virus (HIV). Knowledge of skin testing procedures and symptoms was generally good, and most reported health care providers as the main source of information. Over half reported concern about catching tuberculosis and over 80% favored controls to ensure adherence, in particular directly observed therapy. Higher TB knowledge score (P = 0.0155) and male sex (P = 0.0357) were associated with a favorable attitude toward directly observed therapy. CONCLUSIONS: Health care providers should expand educational messages beyond skin testing. Greater knowledge about tuberculosis may increase acceptance of control measures. Targeted education plus social norms favoring completion of therapy may improve screening and treatment outcomes in this population. PMID- 10383066 TI - The role of tissue studies in facilitating early initiation of antimycobacterial treatment in AIDS patients with disseminated mycobacterial disease. AB - SETTING: The question of whether aggressive investigations are useful in diagnosis and initiation of treatment in AIDS patients with disseminated mycobacterial disease (DMD) is still under debate. OBJECTIVE: To define the role of tissue studies in facilitating early initiation of antimycobacterial treatment and in establishing diagnosis in AIDS patients with DMD. DESIGN: From July 1994 through June 1997, 167 AIDS cases with fever were evaluated by stepwise investigation using a standardized protocol. Data of DMD cases were analyzed to define the role of tissue studies. RESULTS: A total of 40 cases of culture-proven DMD were identified. Antimycobacterial treatment was initiated due to positive acid-fast bacilli smears of sputum in only five cases. In the remaining cases, positive pathologic findings from tissue biopsies (lymph node, bone marrow or liver) facilitated early initiation of treatment in 60% (21/35). In 50% of all cases (20/40), the diagnosis could not have been established if cultures of tissue biopsies had not been performed. Both the pathologic examinations and mycobacterial cultures from liver biopsies had positivity rates of more than 50% (53.8% and 69.2%, respectively). CONCLUSIONS: Tissue studies were useful in facilitating early initiation of treatment and establishing diagnosis at least in half of the AIDS cases with DMD. Liver biopsy is worthwhile if the cause of fever is not discovered using less invasive investigations. PMID- 10383068 TI - A report on home visiting practices conducted in remote districts of Nepal in an NGO-run tuberculosis control programme. AB - To establish the role of home visiting in an NGO-run tuberculosis control programme in Nepal, information was collected on home visits to a cohort of 205 smear-positive patients. Almost one third of new smear-positive cases were visited, either for treatment initiation (n = 33) or for retrieval following non attendance (n = 29); thus 14% of patients required a home visit to ensure treatment completion. It is unlikely that the WHO-recommended target of 85% cure rate would be achieved without defaulter tracing, although a further study comparing home visiting against no visiting would be necessary to assess the contribution that this activity makes to improving treatment outcomes. PMID- 10383069 TI - Simultaneous infection with two strains of Mycobacterium tuberculosis identified by restriction fragment length polymorphism analysis. AB - Simultaneous infection with two different strains of Mycobacterium tuberculosis has been demonstrated using phage typing. We report here the first case of mixed infection identified using IS6110-based genotyping of M. tuberculosis. The patient was diagnosed with pulmonary tuberculosis in February, 1991. The initial isolate of M. tuberculosis had two different genotype patterns (dark 7-band and light 14-band patterns). However, in a repeat isolate obtained several months later, only the 14-band pattern was visible. Exogenous reinfection and laboratory cross-contamination were unlikely because both genotype patterns were unique in the San Francisco database which includes over 1300 isolates of M. tuberculosis. This case demonstrates the importance of identifying mixed infections in the study of the molecular epidemiology of tuberculosis. Mixed infections could be confused with exogenous reinfection or laboratory cross-contamination, and important epidemiologic connections could be missed. PMID- 10383070 TI - Is cough alone adequate to screen HIV-positive persons for tuberculosis preventive therapy in developing countries? AB - Although the efficacy of isoniazid in the prevention of tuberculosis in HIV infected persons with a positive tuberculin skin test is proven, several feasibility issues remain unanswered. In resource poor settings where a chest radiograph may not be readily available, the question of whether cough alone is an adequate screening tool needs to be considered. We analysed screening data collected as part of an isoniazid efficacy study. Although the study was not designed specifically to answer this question, the data suggests that cough alone may be inadequate for screening patients for potential tuberculosis preventive therapy, and that a chest radiograph may be necessary. Feasibility studies are needed. PMID- 10383071 TI - Molecular characterization of Mycobacterium bovis BCG: Romanian sub-strain. AB - The Romanian sub-strain used for BCG vaccine production was characterized by polymorphic GC-rich repetitive sequence (PGRS) restriction profile and IS6110 detection. For comparison, the Pasteur and Moscow Mycobacterium bovis BCG sub strains, M. bovis AN5, and M. tuberculosis H37Rv were analyzed. The BCG sub strains showed the same restriction profile for PGRS after Pvu II, BamH I and Sal I digestion. They could be distinguished from the strains M. bovis AN5 and M. tuberculosis H37Rv after Pvu II and Sal I digestion. Sal I can also distinguish between M. bovis AN5 and M. tuberculosis H37Rv. Digestion with Pvu II, Sal I, Sca I, Mlu I and BamH I gave identical profiles of IS6110 hybridization for the Romanian and Pasteur BCG substrains. PMID- 10383072 TI - HIV-associated mycobacteraemia in West Africa. PMID- 10383073 TI - Controlling tobacco use within prisons. PMID- 10383074 TI - Plasma post-heparin lipase activities in the HERITAGE Family Study: the reproducibility, gender differences, and associations with lipoprotein levels. HEalth, RIsk factors, exercise Training and GEnetics. AB - OBJECTIVES: Examine the reproducibility of plasma lipid and lipoprotein measurements in the HERITAGE Family Study. DESIGN AND METHODS: In a sample of 379 subjects (191 men and 188 women), reproducibility was determined for lipids, lipoproteins (done on two occasions) and post-heparin lipase assays using an Intracenter Quality Control study by generating split samples from an additional 60 subjects (35 men and 25 women), which were assayed in a blind fashion by the lipid core laboratory. Reproducibility was estimated using intraclass correlation coefficients (ICC) for the selected variables. Analytical error (ANER) and coefficient of variation (CV) were also calculated. Day-to-day variation for 10 variables including plasma cholesterol and triglycerides (TG), HDL-cholesterol and its subfractions HDL2-cholesterol and HDL3-cholesterol, LDL-cholesterol and VLDL-cholesterol, as well as apoprotein (apo) A-I, apo B, and LDL-apo B were assessed. RESULTS: In the HERITAGE study, all lipid and lipoprotein variables had ICC above 0.79. Plasma VLDL-cholesterol (31 %) and TG (23%) levels, which are well known to be highly variable from one day to another, had CVs greater than 20%. Other variables had CVs lower than 10% except for HDL2-cholesterol which reached 16%. In the intracenter reliability sub-study, the measurement errors were found to be low except for HDL2-cholesterol. For the lipases, the reproducibility of repeated samples was very high, with ICC over 0.95. The within assay CV corresponded to 2.1 and 5.3% for hepatic lipase (HL) and lipoprotein lipase (LPL), respectively, whereas the between-assay CV reached 8-12% for HL and about 15% for LPL. Due to the complexity of these two assays, the results are considered to be quite satisfactory. CONCLUSIONS: The reproducibility of plasma lipid and lipoprotein measurements, as well as of post-heparin lipase activities, is good in the multicenter HERITAGE Family Study. In addition, the well documented gender difference in the plasma lipoprotein profile was confirmed in the present study, women having lower fasting triglyceride and LDL-cholesterol levels than men as well as reduced cholesterol/HDL-cholesterol and increased HDL2 cholesterol/ HDL3-cholesterol ratios compared to men. Results of the present study support the notion that the higher LPL and low HL activities found in women compared to men are important factors contributing to explain gender difference in the lipoprotein profile. However, additional factors not examined in the present study are involved beyond the contribution of post-heparin lipase to the sex dimorphism in plasma lipoprotein levels. PMID- 10383075 TI - Biochemical studies on leukocyte and fibroblast human beta-galactosidase. AB - OBJECTIVES: Some biochemical characteristics of the human leukocyte and fibroblast beta-galactosidase were studied. DESIGN AND METHODS: Leukocyte and fibroblast enzyme activity was determined fluorometricaly using 4 methylumbelliferyl-beta-D-galactoside as artificial substrate. Optimum pH, Km, Vmax and thermostability of the enzyme at 42 degrees C were determined. RESULTS: The leukocyte and fibroblast enzyme has an optimum pH at 4.2, which is in agreement with the lysosomal origin of the enzyme. The Km of the enzyme was 0.62 in leukocytes and 0.67 in fibroblasts, and Vmax was 289.9 nmol/h/mg of protein and 1779.2 nmol/h/mg of protein in the two tissues, respectively. When fibroblast or leukocyte beta-galactosidase was pre-incubated at 42 degrees C, it did not retain its activity because the residual activity after 80 minutes of pre incubation at this temperature was lower than 30% of the initial activity both in leukocytes and fibroblasts. CONCLUSIONS: This was the first study of Km, Vmax and thermostability of beta-galactosidase performed on leukocytes and provided data for a better characterization of the enzyme beta-galactosidase, allowing the improvement of the analytical conditions. PMID- 10383076 TI - A fluorescence method for the determination of plasma susceptibility to lipid peroxidation. AB - OBJECTIVE: We propose a fluorescence kinetics method for monitoring plasma susceptibility to peroxidation. DESIGN AND METHOD: Plasmatic peroxidation was induced by CuSO4 (500 microM), and fluorescence was measured every 30 min. Kinetics were represented by a sigmoidal curve from which it was possible to calculate the latency time (lag-time) and the propagation velocity (slope) of plasma peroxidation. RESULTS: The lag-time monitored by the fluorescence kinetics method corresponded to the formation of thiobarbituric acid reactive substances, and to progressive depletion of polyunsaturated fatty acids and alpha-tocopherol. The mechanism of reaction appeared to be dependent upon plasmatic hydroperoxides, and independent of oxygen radicals. Plasma storage is possible for at least two months at -80 degrees C, and reproducibility of the method is very good. CONCLUSIONS: Fluorescence kinetics provide a highly comprehensive picture of plasma susceptibility to peroxidation in comparison with the conventional measurements of anti- and pro-oxidant ratios. PMID- 10383077 TI - A comparison of ELISA assays as routine diagnostic test for detection of autoantibodies against extractable nuclear antigens. AB - OBJECTIVE: In an analytical evaluation, commercially available ELISA test kits for detection of antibodies directed against extractable nuclear antigens (ENA) were compared with the currently used combination of counterimmunoelectrophoresis and immunoblotting. DESIGN: Three screening ELISAs and two typing ELISAs were tested. These methods were fairly simple, easy to perform and "user friendly," because most of the reagents were ready to use. RESULTS: The agreement with the current methods was good, but the screening as well as typing ELISAs proved to be more sensitive, especially with regard to detection of SS-A auto-antibodies. The cut-off range of one screening assay was not well established and one typing assay suffered from problems with inaccuracy of package insert, purity of antigen and standardisation of reactivity (possibly caused by differences in amount of coated antigen). The other three ELISAs were reliable and sensitive for detection of ENA auto-antibodies. CONCLUSIONS: The ELISA ENA screen assays ENA-LISA polyvalent and Milenia ENA screen and typing assays ENA-LISA are reliable and sensitive for detection of autoantibodies in clinical specimens without substantial false negatives. PMID- 10383078 TI - Elimination of superoxide dismutase interference in fructosamine assay. AB - OBJECTIVE: Superoxide dismutase (SOD) (EC 1.15.1.1) is reported to decrease the reduction of nitroblue tetrazolium (NBT) in the fructosamine assay. The study was undertaken to find a method to eliminate this interference. DESIGN AND METHODS: We studied the NBT reduction in the presence and absence of added bovine erythrocyte SOD during fructosamine assay. Formation of reduced NBT decreased with the increasing concentration of SOD. Various inhibitors of SOD were experimented with for effectively eliminating this interference. RESULTS: Cyanide eliminates the interference due to SOD, but is unsuitable because in it's presence glucose becomes reducing under the conditions of fructosamine assay. SOD inhibitors such as EDTA and Azide did not eliminate the effect of SOD. Guanidine. HCl gives opalescence in the reaction mixture. Addition of 2M HCl to the serum and incubation at 37 degrees C for 10 min eliminated the effect of added SOD (70 kU/L). The correlation between deltaA10-20 min of serum treated with HCI in presence and absence of added SOD is y = 0.9011x + 0.01055 with r = 0.9789 and S.E. (y) 0.007686. CONCLUSION: SOD does not interfere in globin bound fructosamine assay as acid-acetone treatment in preparation of heme free globin inhibits SOD. Pretreatment of serum with HCl can satisfactorily eliminate the interference due to SOD in fructosamine assay. The acid treatment could be used to inhibit SOD in various other reactions that are followed with NBT reduction. PMID- 10383079 TI - Elevated lipid peroxidation and disturbed antioxidant enzyme activities in plasma and erythrocytes of patients with uterine cervicitis and myoma. AB - OBJECTIVES: We investigated whether oxidative stress is associated with human uterine cervicitis and uterine myoma. DESIGN AND METHODS: We measured lipid peroxidation and antioxidant enzymes in plasma and erythrocytes of cervicitis patients and myoma patients in comparison with matched controls. Thiobarbituric acid-reactive substances (TBARS), a measure of lipid peroxidation, were determined in plasma; glutathione peroxidase (GSHPx) and catalase in erythrocytes; and superoxide dismutase (SOD) in both plasma and erythrocytes. RESULTS: We showed that plasma TBARS were significantly higher (p < 0.05) in both cervicitis patients and myoma patients than in controls. Plasma TBARS were significantly (and negatively) correlated with plasma and erythrocyte T-SOD activities in cervicitis patients only. Plasma T-SOD activity was significantly lower in both groups of patients than in controls whereas erythrocyte T-SOD activity was only significantly lower in myoma patients. The lowered plasma T-SOD activity in the cervicitis patients was attributed to decreased Mn-SOD activity whereas the lowered plasma T-SOD activity in myoma patients was attributed to decreased activities of both Cu,Zn-SOD and Mn-SOD. Erythrocyte GSHPx activity was 14% higher (p < 0.05) in cervicitis patients and 11% lower (p > 0.05) in myoma patients than in controls; catalase activity was 10% higher (p > 0.05) in cervicitis patients and 13% lower (p > 0.05) in myoma patients than in controls. Neither erythrocyte GSHPx nor catalase activity was significantly correlated with plasma TBARS. CONCLUSIONS: The elevated lipid peroxidation and disturbed antioxidant enzyme activities demonstrate the potential of oxidative injury in patients with uterine cervicitis and myoma. PMID- 10383081 TI - Association between AGT T235 variant and microalbuminuria in Canadian Oji-Cree with type 2 diabetes mellitus. AB - OBJECTIVE: To assess the association between the common variation in the gene encoding angiotensinogen, AGT, and the presence of microalbuminuria in Canadian Oji-Cree with type 2 diabetes mellitus. RESEARCH DESIGN AND METHODS: We compared the frequencies of the AGT promoter and M235T polymorphisms among three subgroups of adult Oji-Cree: 50 subjects who had type 2 diabetes with microalbuminuria, 6 subjects who had type 2 diabetes without albuminuria and 302 non-diabetic, normotensive subjects. RESULTS: We found the AGT T235 allele was present at a significantly higher frequency, and that T235/T235 homozygotes were significantly more prevalent, among the subjects who had type 2 diabetes with microalbuminuria than among the subjects in the other two groups. CONCLUSIONS: The findings suggest that the AGT T235 allele is a determinant of the nephropathy susceptibility related to type 2 diabetes in these aboriginal Canadians. PMID- 10383080 TI - Differential effects of low-level lead exposure on the natural isozymes of erythrocyte 5'-nucleotidase. AB - OBJECTIVE: Erythrocyte pyrimidine 5'-nucleotidase (Pyr-5'-N) is highly sensitive to heavy metal inactivation in vitro and in vivo, and a number of studies have verified its usefulness as a biomarker of acute and chronic lead exposures. Retrospective and prospective studies attempted to determine whether the known linearity of Pyr-5'-N inhibition by lead concentrations above 40 microg/dL whole blood might continue into the lowest range of exposures now considered to be toxic in children (<10-20 microg/dL), thereby extending its value as a biomarker of lead exposure. DESIGN: Activities of Pyr-5'-N and a lead-insensitive isozyme, deoxyribonucleotidase (d-5'-N), were compared to blood lead and free erythrocyte protoporphyrin (FEP) concentrations. RESULTS: Pyr-5'-N activities in erythrocytes from 70 children displayed an inverse linear correlation with whole blood lead of 1-35 microg/dL, whereas d-5'-N did not correlate. There was no apparent minimum threshold for Pyr-5'-N inhibition by lead. CONCLUSIONS: Linearity of Pyr-5'-N inhibition by lead extends throughout the range of clinical concern in pediatric cases. Pyr-5'-N/d-5'-N activity ratios may provide an even more sensitive, internally controlled biomarker of low-level lead overburden, since both isozymes vary comparably in activity as a function of reticulocytosis and mean red cell age. PMID- 10383082 TI - Assessment of liver function by the MEGX test in patients with schistosomiasis and cirrhosis. AB - OBJECTIVES: The aim of this study was to determine the effect of schistosomiasis infection on hepatic function in Egyptian patients with posthepatitic cirrhosis. DESIGN AND METHODS: Hepatic function, was assessed in 66 Egyptian patients, with (n = 30) and without (n = 36) schistosomal liver fibrosis due to Schistosoma mansoni and in 20 healthy controls, using the monoethylglycinexylidide (MEGX) test. Serum MEGX concentrations were measured before and 5, 15, 30, 60, 120, and 180 min after a lidocaine bolus. The sero-prevalence of antibodies to hepatitis C was also determined in the patients. RESULTS: MEGX test results were significantly lower in patients than in controls at all time points. MEGX test results declined with advancing Child Class. Receiver operating characteristic (ROC) curve analysis revealed the following areas under the ROC curves for discrimination of Child Class C from Child Classes A/B: 30 min, 0.762; 60 min, 0.743; 120 min, 0.731; 15 min, 0.728; 180 min, 0.728; 5 min, 0.602. Schistosomiasis infection had no influence on MEGX test results when cirrhotic patients with (Schisto+) and without (Schisto-) schistosomiasis were compared. While the prevalence of the hepatitis B surface antigen was only 16.7% (Schisto-) and 26.7% (Schisto+), there was an extremely high sero-prevalence of antibodies to hepatitis C (HCV) in both groups: 88.9% (Schisto-) and 73.3% (Schisto+). CONCLUSIONS: The association of schistosomal liver fibrosis with cirrhosis does not additionally influence MEGX formation. In addition, HCV rather than schistosomiasis infection must be considered as a major cause for the progressive liver disease in these patients. PMID- 10383083 TI - Corticosteroid-binding globulin and free cortisol in the early postoperative period after cardiac surgery. AB - OBJECTIVES: To characterize concentrations of corticosteroid-binding globulin (CBG), total and free serum cortisol, and free urinary cortisol in patients during the postoperative period of cardiac surgery. DESIGN AND METHODS: In 24 patients serum was sampled on the first and second postoperative day after cardiac surgery (21 procedures with thoracotomy, 3 thoracoscopic procedures); urine was collected for two 10-h periods (8 P.M. until 6 A.M.) on the respective postoperative days. Total serum cortisol and free urinary cortisol were measured with an automated chemiluminescence assay (analysis of urine after extraction with dichloromethane), and CBG using a coated-tube RIA. Free serum cortisol was calculated from the concentrations of total serum cortisol and CBG as described previously. Thirty healthy volunteers were studied as controls. RESULTS: CBG was reduced to about one-half of the normal concentration on both postoperative days. Whereas total cortisol was about two-fold increased on the first postoperative day compared to controls extremely high concentrations of free serum cortisol were calculated from CBG and total cortisol [median 136 nmol/L (interquartile range 100-185); controls 21.8 nmol/L (interquartile range 16.9-29.8)]. On the second postoperative day, median total serum cortisol was within the interquartile range of the controls, free serum cortisol in contrast was still two-fold increased. Free serum cortisol and free urinary cortisol were significantly correlated (r = 0.60). CONCLUSIONS: Extremely high concentrations of free serum cortisol are typically found in the postoperative period of cardiac surgery; under these conditions the mere consideration of total cortisol does not appropriately display the activation of the adrenal cortex. PMID- 10383084 TI - Apolipoprotein E genotype and plasma levels in coronary artery disease. A case control study in the Italian population. AB - OBJECTIVES: To investigate the role of the apolipoprotein B and apolipoprotein E polymorphisms in coronary artery disease (CAD) susceptibility in the Italian population and their relation to plasma lipid and apolipoprotein levels. METHODS: APOB (APOB Xbal, EcoRI, Ins/Del), and APOE (APOE Cfol) polymorphisms were analyzed in 150 male CAD patients and 110 matched controls. In the same subjects plasma lipid, apoB, and apoE levels were measured. RESULTS: No differences in the distribution of the APOB polymorphisms were observed between patients and controls. Among patients the number of e*4-carriers was significantly higher than in controls. e*4-carriers were more frequent among the hypertensive patients and had a higher systolic blood pressure (p = 0.007) than the non-e*4 carriers. The APOB Xbal polymorphism was found to influence the distribution of HDL cholesterol. Patients showed significantly lower levels of apoE (39.29 mg/L) than controls (54.32 mg/dL) and the lowest concentrations were associated to the E4/E3 and E4/E4 genotypes. CONCLUSION: Quantitative data are consistent with the hypothesis that apoE has an anti-atherosclerotic role and suggest that the apoE quantitation could be a useful parameter for defining cardiovascular risk. e*4 allele appears to be a risk factor for CAD in the Italian population and could act by its association with low apoE levels. PMID- 10383085 TI - Genetic determinants of heritable venous thrombosis: genotyping methods for factor V(Leiden)A1691G, methylenetetrahydrofolate reductase C677T, prothrombin G20210A mutation, and algorithms for venous thrombosis investigations. AB - OBJECTIVES: To implement cost effective and clinically relevant thrombophilic genotyping and homocysteine analysis in our coagulation laboratory. METHODS: We describe genotyping assays for three of the genetic defects associated with hereditary thrombosis: factor V(Leiden) A1691G, methylenetetrahydrofolate reductase C677T, and prothrombin gene G20210A. A second confirmatory assay for factor V(Leiden) using allele specific oligonucleotide polymerase chain reaction is also presented. We suggest an algorithm for the rational integration of the traditional assays routinely used to investigate venous thrombosis with genotyping and plasma homocysteine measurements. RESULTS: These polymerase chain reaction based assays were designed to be performed under identical reaction conditions, permitting simultaneous setup, amplification, digestion, and analysis. CONCLUSIONS: The three genotyping assays presented are robust and relatively easy to perform. Use of an algorithm will ensure efficient resource utilization and minimize unnecessary testing. PMID- 10383086 TI - Multicenter evaluation of an amperometric immunosensor for plasma fatty acid binding protein: an early marker for acute myocardial infarction. PMID- 10383087 TI - Improved nonisotopic PCR-SSCP for screening of p53 mutations. PMID- 10383088 TI - Performance characteristics of the creatine kinase-MB mass assay on the Vitros ECi analyzer. PMID- 10383089 TI - Assessment of induced rat mammary tumour response to chemotherapy using the apparent diffusion coefficient of tissue water as determined by diffusion weighted 1H-NMR spectroscopy in vivo. AB - Chemosensitivity of N-methyl-N-nitrosourea-induced rat mammary tumours treated with 5-fluorouracil at a dose of 100 mg kg(-1) i.p. was assessed by using diffusion-weighted 1H-MRS to measure the average diffusion coefficient (ADC) of water in the tumour tissue. ADC measurements prior to any therapy correlated positively with necrotic fraction. Tumours with low initial ADC (< 0.95 x 10(9) m2 s(-1)) showed an increase in ADC 7 days after treatment, whereas tumours with a high initial ADC (> 1.2 x 10(9) m2 s(-1)) showed a decrease. All tumours decreased significantly in volume (P < 0.05) 2, 5 and 7 days after treatment. At day 7 post-treatment, tumours with a high pre-treatment ADC started to regrow. The initial ADC value, as well as changes after treatment predict tumour chemosensitivity, which could be clinically relevant. PMID- 10383090 TI - In vitro image characteristics of an abdominal aortic stent graft: CTA versus 3D MRA. AB - Percutaneous stent-grafting is increasingly employed as a less invasive alternative to surgery for the treatment of infrarenal abdominal aortic aneurysms. It requires long-term imaging follow-up, to document the structural integrity of the device, to exclude perigraft channels and endograft leakages, as well as the shrinkage of the aneurysmal sac. The expectation of severe stent induced artifacts and safety concerns have prevented 3D MRA from being used. The purpose of this in vitro study was to investigate the imaging characteristics of a bifurcated stent graft with 3D MRA (3D Fourier transform fast spoiled GRE) at 1.5 T in comparison to those of CTA. Measurement of the stent wall thickness and luminal diameter were made on a agar gel embedded stent graft at five locations on both CTA and MRA images. The stent graft was depicted as a dark ring on MR images. Wall thickness measurements at the five locations of the stent graft overestimated the true stent thickness, while luminal diameters were slightly underestimated. Measurement differences between MR and CT were not statistically significant (P = 0.67; P = 0.85). Artifacts emanating from the platinum markers were considerably less severe on the MR-images. A wider area of signal loss was seen only at the insertion of the iliac stent leg into the aortic stent portion due to the overlap of two radio-opaque platinum markers. 3D MRA images should permit a comprehensive assessment of the arterial lumen, and of perivascular tissues. PMID- 10383091 TI - Identification of cerebral acetone by 1H-MRS in patients with epilepsy controlled by ketogenic diet. AB - Ketogenic diet (KD) is highly effective in controlling epileptic seizures in children. One of the mechanisms postulated, the accumulation of ketone bodies, acetoacetate (AcAc) and/or betahydroxybutyrate (beta-OHB) in the brain, would be detectable by non-invasive proton magnetic resonance spectroscopy (1H-MRS). 1H MRS was performed in occipital cortical grey matter in 14 epileptics (E); ages 3.8-48.3 years), seven KD and seven without, and 16 healthy age-matched subjects. One E was examined before and after KD. A singlet resonance (sigma = 2.22 ppm), distinct from AcAc (sigma = 2.26 and 3.46 ppm), and identified as acetone was present in all spectra of children on KD (nine spectra in five children; concentration 0.7 +/- 0.2 mM). This resonance was absent from Control and E without diet. AcAc and beta-OHB, which were not detectable in KD brain, were found in urine or blood of all KD. Seizures were well controlled in all E in whom acetone was detected. Two of seven E, both adults, were seizure-free without detectable acetone. Cerebral acetone may contribute to seizure control in KD, but is unlikely to be the sole mechanism. PMID- 10383092 TI - The modular (twin) gradient coil--high resolution, high contrast, diffusion weighted EPI at 1.0 Tesla. AB - The modular (twin) gradient coil is a novel and effective approach to obtaining flexible and high power gradient performance without (i) peripheral nerve stimulation and (ii) the need for resonant or expensive high voltage gradient power supply units (PSU). This whole-body gradient system contains, on the same former, two independent sets of gradient coils, the conventional coil set and the short-body coil set. Each gradient axis is able to operate, independently, in any one of three modes. The third, combined, mode is realized by connecting the conventional coils and short-body coils in series. Through careful design of the shape and size of the linear volume of each mode of operation, the modular gradient coil is able to utilize the power, from a single gradient PSU, more efficiently and more appropriately, as determined by the application. In the short-body and combined modes the gradient fields are linear over a volume suitable for whole head/neck, liver and cardiovascular applications. In the conventional mode, a reduced performance is possible but over a much larger (conventional) imaging volume. Utilizing a semi-conductor switching arrangement it is possible to switch between modes in as little as 1 ms. By mixing different modes of operation on different gradient axes it is possible to utilize more efficiently, and safely, the properties of gradient performance best suited to the sequence requirements. Diffusion weighted EPI (DW-EPI) is a particular technique that demands the extremes of gradient system performance in terms of both amplitude and slew rate. DW-EPI has been implemented, using the modular gradient system, on a 1.0 Tesla whole-body MRI system. The preliminary results presented here serve to illustrate the advantages of the modular gradient coil in of itself as well as the direct benefits it provides for DW imaging at 1.0 Tesla. PMID- 10383093 TI - BOLD-MRI in ten patients with coronary artery disease: evidence for imaging of capillary recruitment in myocardium supplied by the stenotic artery. AB - Changes of myocardial oxygenation can be studied by measurements of the apparent transverse relaxation time T2*, which is correlated with the oxygenation state of hemoglobin. In this study, ten patients with coronary artery disease (CAD) underwent blood oxygenation level dependent (BOLD) T2* measurements using a segmented gradient echo pulse sequence with ten echoes. T2* measurements were performed in a single short-axis slice of the heart at rest and under pharmacological stress with dipyridamole (DIP), which increases myocardial blood flow. For comparison, all patients underwent X-ray angiography and stress echocardiography within 4 days after the MR exam. In one patient, MR examination was repeated 10 weeks after percutaneous transluminal coronary angioplasty (PTA). In the differential T2* maps, expected ischemic areas of myocardium were identified in six patients. In these regions, T2+ values (30 +/- 8 ms) were significantly reduced when compared to the remaining myocardium (48 +/- 9 ms, P < 0.01). In four patients, the myocardial region of interest could not be assessed owing to severe susceptibility artifacts in the ischemic region. The success of the PTA treatment could be visualized from a more homogeneous DIP induced increase in T2* within the ischemic myocardium (from 26 +/- 1 to 29 +/- 1 ms before PTA versus 26 +/- 1 to 31 +/- 4 ms after PTA, P < 0.001). PMID- 10383095 TI - The effects of selenium deficiency on differentiation, degradation, and cell lysis of L8 rat skeletal muscle cells. AB - We investigated the effects of selenium (Se) deficiency on differentiation, protein degradation, and cell lysis in cultured skeletal muscle cells, using L8 rat skeletal muscle cells cultured in serum-free (SF) medium to induce differentiation and to maintain myotubes. Creatine kinase activity was reduced (p < 0.05) by approximately 15% without Se supplementation for 96 h. Confluent myoblasts were treated with SF media with four different levels of vitamin E (0, 10, 35, and 100 microM) in the absence and presence of Se (0 and 0.25 microM, respectively). After 96 h, vitamin E at a high dose (100 microM) was effective in the prevention of the decrease of differentiation caused by Se deficiency (p < 0.05). Following differentiation, the effects of three Se concentrations (0, 0.25, and 2.5 microM) on degradation of proteins as assessed by release of 3H labeled free amino acids secreted into the media were studied. Selenium supplementation did not affect (p > 0.05) total protein degradation. However, Se deficiency increased (p < 0.05) lactate dehydrogenase released from lyzed dead cells. The results indicate that Se is required to maintain an optimal rate of muscle cell differentiation and health of myotube cultures. PMID- 10383094 TI - Cervical carcinoma: standard and pharmacokinetic analysis of time-intensity curves for assessment of tumor angiogenesis and patient survival. AB - Since detailed knowledge regarding the pathophysiological properties which in turn are responsible for differences in contrast enhancement--remain fairly undetermined, it was the aim of this study (i) to examine the association of standard and pharmacokinetic analysis of time-intensity curves in dynamic MRI with histomorphological markers of tumor angiogenesis (microvessel density [MVD]; vascular endothelial growth factor [VEGF]); and (ii) to determine the ultimate value of a histomorphological and a dynamic MRI approach by correlation of those data with disease outcome in patients with primary cancer of the uterine cervix. Pharmacokinetic parameters (amplitude A, exchange rate constant k21) and standard parameters (maximum signal intensity (SI)-increase [SI-I] over baseline and steepest SI-upslope per second [SI-U/s]) were calculated from contrast-enhanced dynamic MR imaging series in 37 patients with biopsy-proven primary cervical cancer. On the surgical whole mount specimens, histomorphological markers of tumor angiogenesis (MVD, VEGF) were compared with similar sized and positioned regions-of-interest (ROIs) on the MRI-derived parameters. For MRI and histomorphological data, Kaplan-Meier survival curves were calculated and compared using log-rank statistics. A significant association was found between MVD and amplitude A (P < 0.01) and SI-I (P < 0.05). No significant relationships were observed between the VEGF expression and all dynamic MRI parameters. Kaplan Meier curves based on k21 and SI-U/s showed that tumors with high k21 and SI-U/s values had a significantly (P < 0.05, 0.001, respectively) worse disease outcome than tumors with low k2, and SI-U/s values. None of the histomorphological gold standard markers for assessing tumor angiogenesis (MVD, VEGF) had any significant power to predict patient survival. It is concluded that (1) the pathophysiological basis for differences in dynamic MRI is MVD but not VEGF expression; (2) a functional, dynamic MRI approach (both standard and a pharmacokinetic analysis) may be better suited to assess angiogenic activity in terms of patient survival than current histomorphological-based markers of tumor angiogenesis; and [3] compared with standard analysis, a simple pharmacokinetic analysis of time-intensity curves is not superior to assess MVD or patient survival. PMID- 10383096 TI - Effect of selenium supplementation in asthmatic subjects on the expression of endothelial cell adhesion molecules in culture. AB - Endothelial cells play a major role in immunologic reactions, in which cellular adhesion molecules P-selectin, ICAM-1, VCAM-1, and ELAM-1 are important mediators in the recruitment of leukocytes in pulmonary inflammation. Selenium (Se) is known to modulate immunological mechanisms of asthma. The aim of our investigation was to examine whether Se supplementation in cortico-dependent asthmatic patients may modulate adhesion molecule expression in cultured endothelium. Our findings indicated that P-selectin, VCAM-1, and ELAM-1 expression on human umbilical vein endothelial cells stimulated with peripheral blood mononuclear cells obtained from asthmatics before supplementation with Se was significantly higher than from healthy donors (p < 0.05). The production of ICAM-1 showed only slight augmentation. The levels of VCAM-1 and ELAM-1 expression were significantly decreased after 3 mo of Se supplementation (p < 0.05). After 6 mo of intervention period the intensity of P-selectin and ICAM-1 expression was also significantly reduced (p < 0.05 and p < 0.01, respectively). The inhibitory effect of Se on the adhesion molecule expression was studied in cultured endothelial cells after interferon-gamma stimulation. Our data suggest that Se affects the expression of P-selectin, ICAM-1, VCAM-1, and ELAM-1 in a dose-dependent manner and the half-maximal inhibitory concentrations were 3.4, 0.5, 4, and 3.8 microg/mL, respectively. The maximal inhibitions (greater than 80%) were observed in vitro with 10 microg/mL Se (p < 0.01). Regulation of adhesion molecule expression may be an important mechanism through which the inflammation may be controlled. PMID- 10383097 TI - Zinc, copper, and iron metabolism during porcine fetal development. AB - Zinc, copper, and iron levels in maternal and fetal pig tissues and fluids were measured starting on d 30 of gestation and continuing to term (d 114) at 10-d intervals. Fetal hematocrit increased from a low of 19% on d 30 to 32% by d 50, after which it remained above 30% to term. Amniotic fluid zinc, copper, and iron all reached maximal levels by d 60 of gestation. Maternal serum zinc levels fluctuated little during gestation, but fetal serum zinc concentration was significantly elevated above maternal levels during the second trimester. Fetal serum copper levels were significantly lower than maternal values throughout gestation and this was also the case for ceruloplasmin oxidase activity. Maternal serum iron reached its lowest level by d 80 of gestation when rate of transfer of iron to the developing fetuses was high. Fetal serum iron declined throughout gestation, reaching its lowest level on d 100. In general, fetal liver concentrations of zinc, copper, and iron were higher than the corresponding maternal values throughout gestation. Distinct increases were noted for fetal hepatic zinc and copper concentrations during the second trimester of pregnancy and these were accompanied by increases in cytosolic and metallothionein-bound zinc and copper levels. Maternal hepatic iron declined during the second trimester, reaching its lowest point on d 80, indicative of the shunting of maternal iron reserves to fetal tissues. Fetal kidney metal levels did not demonstrate any distinctive developmental patterns with respect to zinc, copper, or iron concentrations, but a general accumulation of each metal was observed as gestation progressed. The results of this study highlight some of the distinct changes occurring in the metabolism of zinc, copper, and iron in both maternal and fetal tissues and fluids during gestation in the pig. PMID- 10383098 TI - Serum extracellular superoxide dismutase activity as an indicator of zinc status in humans. AB - The present study focused on whether serum extracellular superoxide dimutase (EC SOD) activity can be used as a functional indicator of marginal zinc deficiency in humans. Subjects in this study were 444 healthy adults over 30 yr of age living a normal rural life in Kyunggi province, Korea. The mean dietary zinc intake of subjects obtained from one 24-h recall was 6.41 +/- 4.35 mg and the average serum zinc concentration of the subjects was 11.06 +/- 2.44 micromol/L. Subjects were divided into three groups by serum zinc concentrations: adequate (serum zinc >10.7 micromol/L), low (serum zinc 9.0-10.7 micromol/L), and very low (serum zinc <9.0 micromol/L) groups. A total of 50 subjects were selected from the three groups for analysis of EC-SOD activities. The EC-SOD activity of subjects increased with increasing serum zinc concentrations, and the activities of the three groups were significantly different as indicated by the Kruskal Wallis test (p = 0.0239). Also, serum EC-SOD activities were significantly correlated with serum zinc concentrations (r = 0.289, p = 0.04). Serum EC-SOD activities, however, were not significantly correlated to the dietary zinc intakes. In conclusion, these results show that EC-SOD activities are decreased in subjects with low serum zinc concentrations and suggest that EC-SOD activity may be a functional indicator of zinc nutritional status in humans. PMID- 10383099 TI - Plasma and urinary selenium in Saudi Arabian patients with dilated cardiomyopathy. AB - We measured selenium (Se) levels in the urine and blood plasma samples of 72 Saudi Arabian patients with dilated cardiomyopathy (DCM) and 70 control subjects of the same origin. To correct for differences in the hydration state of the subjects, the selenium concentration for each urine sample was normalized by dividing it by the concentration of creatinine (CREAT) in the same sample. The median (and range) of the values found for the concentration of Se in plasma, urine, and normalized concentration in urine for the control subjects was 1.306 (0.66-2.50) microM, 0.478 (0.05-2.00) microM, and 56.7 (10.6-426.5) microM Se/M CREAT, respectively, whereas, for the patients, it was 1.246 (0.53-2.45) microM, 0.39 (0.05-1.90) microM, and 75.1 (4.9-656.2) microM Se/M CREAT, respectively. Additionally, the patients were separated into three subgroups according to the severity of their disease state as judged by NYHA procedure, and were then compared to the control group. Only group 4 (the most severe state of the disease) had a significantly lower concentration of urinary Se than the control group. However, the difference became nonsignificant when normalized for CREAT levels. There was no significant difference in the plasma Se levels between the controls and any of the patient groups. As the plasma Se in the control group and in the DCM patients both fell on the low end of the "normal" range, with the patients being marginally lower than the controls, there is no firm evidence from this study to suggest that Se is related to the high incidence rate of DCM found in Saudi Arabia. PMID- 10383100 TI - Study of chemical species of iodine in human liver. AB - The distribution and chemical species of iodine in various subcellular fractions of human liver were studied by using epithermal neutron activation analysis combined with chemical and biochemical separation techniques, such as gradient centrifugation and gel chromatography. It was found that the total iodine content orders in various subcellular fractions is as follows: nuclei > cytosol > mitochondria > lysosome > microsome. In the lysosomal fraction, iodine is mainly bound to macromolecules, whereas in the nuclei and mitochondrial fractions, mainly with lower-molecular-weight organic compounds. In the cytosol fraction, iodine is combined with three proteins, in which iodine is chiefly bound with mid and high-molecular-weight proteins. PMID- 10383101 TI - Cyclosporine and tacrolimus (FK506): a comparison of efficacy and safety profiles. AB - Multicenter clinical trials conducted in the United States and Europe to compare the efficacy and safety of cyclosporine with tacrolimus (FK506) have demonstrated comparable long-term patient survival and graft survival in liver and renal transplant recipients. Importantly, treatment with tacrolimus was associated with reductions in the incidence and severity of acute rejection episodes. However, tacrolimus-based therapy was also associated with increased toxicities in comparison to conventional cyclosporine-based therapy. It is becoming increasingly accepted that earlier trials may have employed high or supratherapeutic doses of tacrolimus and may have been unbalanced with respect to study design. In addition, these pivotal comparative trials were performed with the original formulation of cyclosporine, and not the cyclosporine microemulsion preparation. This critical review of the literature focuses on the United States and European tacrolimus multicenter clinical trials and examines the efficacy and safety of the two primary immunosuppressants, cyclosporine and tacrolimus, obtained in these and other studies. The preliminary findings of ongoing studies comparing the efficacy and safety of the improved formulation, cyclosporine microemulsion, with tacrolimus are also discussed. The overall efficacy of the two agents appears to be similar. The safety profile shows differing toxicities of the two medications. The availability of these two immunosuppressants allows the clinician improved options when choosing an immunosuppressive regimen in solid organ transplantation. PMID- 10383102 TI - Presence of a nitric oxide synthase inhibitor in the graft efflux during reperfusion in human liver transplantation. AB - Involvement of the nitric oxide (NO) system in complications following human orthotopic liver transplants (OLT) has been reported, but the contribution of the graft to the modulation of the NO system during reperfusion in normal OLT has not been characterized. We have studied the contribution of the graft efflux to the modulation of the NO system in 20 consecutive OLT. We evaluated its effects on isolated vascular reactivity of the rabbit and on rat cultured macrophages stimulated with lipopolysaccharide (LPS). In none of the donor liver biopsies was expression of inducible NO synthase (iNOS) activity by Northern or Western blot analysis found. Graft efflux after the onset of liver reperfusion, but not pre transplant patient plasma, reversibly inhibited the acetylcholine-induced relaxation of norepinephrine-contracted rabbit aortic rings. Moreover, graft efflux reversibly inhibited NO production in rat macrophages treated with LPS, as evidenced by both a decrease in nitrite plus nitrate formation and a decrease in the production of [14C]citrulline from [14C]arginine. Addition of a 10% dilution of graft efflux to cultured rat macrophages incubated with LPS increased iNOS mRNA levels, suggesting direct inhibition of the enzyme but not of its expression. These results cannot be ascribed to the depletion of arginine the iNOS substrate since they can be reproduced even in the presence of an excess (10 mM) of exogenously added arginine. No correlation was found between the iNOS inhibitory activity in each sample and the corresponding clinical parameters related to either the graft function after the OLT or the existence of post reperfusion syndrome. Our results indicate the existence of a soluble factor in the graft efflux from human OLT that reversibly and unspecifically inhibits NOS activity. Its involvement in the physiology and/or pathology of human liver diseases deserves further study. PMID- 10383103 TI - Factors influencing the potential organ donation: a 6-yr experience of the New Jersey Organ and Tissue Sharing Network. AB - The demand for organ transplants in the United States is increasing by 16% every year. Unfortunately, organ donation figures are not increasing at the same rate. Factors that influence the process of organ donation in New Jersey were analyzed. METHODS: A retrospective study in which the charts of actual and potential organ donors identified by the New Jersey Organ and Tissue Sharing Network (OTSN) between January 1990 and December 1995 were reviewed. Potential donors who were not identified by the OTNS or the United Network for Organ Sharing (UNOS) were not considered because no data relative to these cases were available. The conversion ratio (CR) between actual donor from potential donor was determined. A statistical analysis of the data was performed using multivariate regression logistic analysis. RESULTS: Organ donation increased, both in the male and female population, by 14% over the last 6 yr. The 0-5-yr age group experienced an increment in CR from 7.7 to 37.7% (p < 0.001). All other age groups had a continuous improvement, but a statistically significant increase over time was not observed. The CR of all races increased over the 6-yr study period. The Afro American population donated significantly less than the white population (32.1 vs. 59.9%) (p < 0.001). The three transplant centers in New Jersey had a CR less than that seen in the non-transplant centers (38.1 vs. 44.1%). The number of total donations (78.7 vs. 21.3%) was significantly greater in the non-transplant centers (p < 0.001). Moreover, the number of lost donors was higher at transplant centers (p < 0.001). Over the 6-yr period, the difference between donations coming from non-urban (70.8%) versus urban areas (29.27%) was highly significant (p < 0.001). Traumatic deaths were associated with a greater CR (55.3%) than all other causes of death. The CR for donors dying as a result of motor vehicle accidents (MVA) (p < 0.001), penetrating trauma, and child abuse all increased. Level II trauma centers had a better CR (53.7%) than level I centers (48.4%) and non-trauma centers (51.1%). The donation rate was similar for level II and non trauma centers (60%). CONCLUSIONS: The organ donation rate in New Jersey is not sufficient to meet the needs of organ recipients in New Jersey. Pediatric donations increased considerably, specifically from child abuse. MVA deaths are associated with the greatest CR. Urban areas have a worse CR than non-urban areas, even if they are associated with transplant or trauma PMID- 10383104 TI - The prognostic importance of severity and type of post-transplant proteinuria. AB - Proteinuria, developing after renal transplantation may influence allograft and patient outcomes. This study aimed to investigate the effect of proteinuria on patient and allograft survival. Among 514 patients, 56 (11%) patients with good allograft function and proteinuria were evaluated retrospectively. Patients with proteinuria were classified as group P (20 patients with permanent proteinuria, Male/Female: 16/4) and group T (36 patients with temporary proteinuria, M/F: 29/7) according to the type of proteinuria. Also, considering the amount of proteinuria, patients were classified as group M (32 patients with massive proteinuria, M/F: 29/3) and group NM (24 patients with non-massive proteinuria, M/F: 16/8). The mean time interval between transplantation and appearance of proteinuria was 23.7 months (range 0-121 months) and no difference was found between groups. Two- and 5-yr allograft survival rates were found to be 85 and 80% in group M, and 95 and 82% in group NM. respectively (p = 0.24). In terms of type of proteinuria, 2- and 5-yr allograft survival rates were found to be 70 and 58% in group P and 92 and 87% in group T, respectively. The difference between groups P and T was found to be statistically significant (p = 0.02). Most (85%) of the patients with permanent proteinuria also had massive proteinuria. In conclusion, we found a significant relation between type and severity of proteinuria. The type of post-transplant proteinuria had a stronger effect on allograft outcome than the severity of proteinuria. PMID- 10383105 TI - A randomized trial of surgical antimicrobial prophylaxis with and without vancomycin in organ transplant patients. AB - BACKGROUND: Gram-positive organisms, including vancomycin-resistant enterococci (VRE), have emerged as major pathogens on the organ transplant service at our institution. We hypothesized that our use of vancomycin as part of routine surgical prophylaxis increased the risk of VRE colonization and infection; conversely, there was concern that failure to use vancomycin prophylaxis would increase peri-operative morbidity due to gram-positive organisms. METHODS: Renal transplant recipients (n = 88) were randomized to receive either a) vancomycin/ceftriaxone or b) cefazolin; and pancreas transplants (n = 24) to receive either a) vancomycin/gentamicin or b) cefazolin/gentamicin. Stool samples or rectal swabs were obtained for culture for enterococci within 24 h of transplantation and weekly while hospitalized. RESULTS: Enterococci were isolated on stool culture from 38 (34%) of 102 patients at the time of transplantation; 4 (11%) of the isolates were VRE. The percentage of patients who subsequently acquired VRE was low (1-7% per wk) but remained constant during hospitalization. There was no association between new VRE detection and vancomycin use for either prophylactic or therapeutic purposes. Forty-four patients (39%) had a post operative infection with 46% of these infections due to gram-positive organisms; rates were unaffected by prophylactic vancomycin use. Pancreas transplant patients who did not receive vancomycin prophylaxis had a significantly longer initial hospitalization (p = 0.03); however, differences were not statistically significant when total length of stay (LOS) within the first 90 d of transplantation was compared. CONCLUSIONS: Vancomycin surgical prophylaxis does not appear to have an effect on VRE colonization or infection, or on rates of infection with gram-positive bacteria. Elimination of vancomycin prophylaxis in renal transplant patients may be a reasonable part of an overall program to limit vancomycin usage, although as a single measure, its impact may be minimal. Vancomycin surgical prophylaxis may be of greater importance in pancreas transplants. PMID- 10383106 TI - The poor accuracy of indirect measurements of cadaveric donor kidney weights. AB - Reports that examined the issue of whether transplantation of inadequate nephron mass may be a risk factor for long-term allograft failure yielded conflicting results. One of the more accurate methods of estimating glomerular mass is kidney weight. Most of the clinical studies used body surface area (BSA) or kidney length as estimates of kidney weight. To test the hypothesis that indirect measures of kidney weight are accurate estimates of kidney weight, we compared the kidney weight of 41 consecutive cadaveric kidneys to donor BSA, dimensions measured with calipers at the time of transplantation, and dimensions supplied by the Organ Procurement Agency (OPA). Linear regression analysis was used with kidney weight as the dependent variable and BSA, kidney length, or kidney volume as the independent variable. Kidney length measured with calipers was also compared to kidney length supplied by the OPA. Kidney weight had the best correlation with kidney volume and kidney length determined by caliper measurements (r = 0.640 and 0.646, respectively). The regression analysis showed that the correlation of kidney weight with BSA was 0.487. The correlation of OPA provided kidney length with kidney weights was poor (r = 0.410). The linear regression of caliper-measured kidney length versus OPA length yielded a slope of 0.360, instead of an ideal slope of 1. The assumption has been made that kidney weight or a surrogate of kidney weight has an excellent correlation with nephron mass. Some of the variability in studies that attempted to examine the effect of transplanted nephron mass on allograft outcome may be due to inaccurate estimates used for kidney weight. Our data suggest that surrogate measurements of kidney weight may not be accurate. We recommend that measured kidney weight should be used in studies examining the effect of donor renal mass on allograft outcomes. PMID- 10383107 TI - Impact of solid organ transplantation and immunosuppression on fever, leukocytosis, and physiologic response during bacterial and fungal infections. AB - Immunosuppressed solid organ transplant patients may exhibit a blunted response to infection compared to non-transplant patients. To test this hypothesis, we prospectively identified all episodes of bacterial and fungal infection on the in patient abdominal organ transplant service in our hospital, in 1997, and compared them to infected general surgery and trauma admissions treated simultaneously on the same wards. Eighty-two infections occurred in transplant patients versus 463 in non-transplant patients. Transplant patients demonstrated an overall greater physiologic response [Acute Physiology and Chronic Health Evaluation (APACHE II) and Acute Physiology Scores (APS) at the time of infection of 17.0+/-0.7 and 10.3+/-0.6, respectively, vs. 12.2+/-0.4 and 8.0+/-0.3 for non-transplant patients, p < 0.003], with a similar maximum temperature (38.0+/-0.1 vs. 38.2+/ 0.1 degrees C, p = 0.2) and white blood cell (WBC) count (12.1+/-1.0 vs. 13.9+/ 0.4 k/mL, p = 0.08). Upon further analysis of subgroups, patients receiving mycophenolate or azathioprine had significantly lower maximum temperatures (37.9+/-0.2 degrees C) and WBC counts (11.0+/-0.9 k/mL) when compared to non transplant patients, while steroids appeared to have little effect on the systemic inflammatory response. Overall mortality was similar between groups. In general, solid organ transplant recipients exhibit a physiologic response to bacterial or fungal infection (as measured by the APS) at least as great as that seen in non-transplant surgical patients, although mycophenolate and azathioprine appear to slightly depress the ability to respond with fever and leukocytosis. None of these differences appeared to affect overall mortality. PMID- 10383109 TI - Phaeohyphomycosis in kidney transplant patients. AB - Dematiaceous fungi are being increasingly recognized as pathogens in organ transplant recipients. This investigation reports five cases of subcutaneous phaeohyphomycosis that occurred in five kidney transplant recipients in the Renal Unit at the San Vicente de Paul Hospital, Medellin, Colombia. Fungi of the genus Exophiala were isolated in three cases, but the agent was not identified in two cases. Physicians are likely to develop increased awareness of the clinical manifestation of infection with the dematiaceous fungi as the population of immunocompromised hosts continues to grow. PMID- 10383108 TI - Delayed graft function: risk factors and the relative effects of early function and acute rejection on long-term survival in cadaveric renal transplantation. AB - Delayed graft function (DGF) and acute rejection have both been associated with reduced renal allograft survival. In some studies, they have been shown to have an interactive effect. We studied the risk factors for DGF and the relative impact of DGF and rejection on both short- and long-term survival in recipients of cadaveric renal transplants. Data from the Oxford Transplant Centre Database were assessed on 710 cadaver allografts over a 10-yr period, during which time all recipients received cyclosporin-based immunosuppressive protocols. The interaction between DGF and acute rejection was examined using logistic and Cox multivariate regression. Long cold ischaemia time (CIT), sensitisation and older donor age were found to be independent predictors of DGF. The occurrence of DGF resulted in a reduced 5-yr survival (56 vs. 75%). However, the effect of DGF was confined to the first year post-transplant, as there was no significant difference in survival, as measured by half-life (t1/2) of grafts functioning at 1 yr, with DGF alone and a group with good early function (t1/2 = 21.3 vs. 20.0 yr). There was no increase in acute rejection in grafts with DGF. However, the combination of DGF and acute rejection resulted in the worst short-term graft survival (68% at 1 yr, compared to 92.3% in those grafts with no DGF or acute rejection) and this continued over the long term (t1/2 = 10.5 yr). These data suggest that early function is critical to the success of renal transplantation. The effects of DGF are limited to the first year post-transplant. Long-term graft survival may be improved by efforts to limit CITs, particularly for grafts from older donors and sensitised recipients. PMID- 10383110 TI - Pseudoaneurysm of the superior mesenteric artery with an arteriovenous fistula after simultaneous kidney-pancreas transplantation. AB - Vascular complications remain a significant source of morbidity after pancreatic transplantation. We describe a pseudoaneurysm of the superior mesenteric artery (SMA) with an arteriovenous fistula (AVF) involving the SMA and the superior mesenteric vein (SMV) discovered and treated surgically in the second week after kidney pancreas transplantation. The patient experienced pain over the graft, and subsequent radionuclide and Doppler ultrasound scan were suggestive of a pseudoaneurysm in the head of the pancreas. Awaiting confirmatory angiography, the patient became hypotensive and after resuscitation, underwent emergency surgery when a pseudoaneurysm was found in the head of the pancreas. After looping the proximal and distal recipient iliac artery and base of the donor Y vascular graft, the AVF was separated and ligated. The SMV was dissected off the pancreatic head and repaired over a tamponading intraluminal Foley catheter. Graft function was preserved. Based on this experience, an AVF with or without a pseudoaneurysm in the pancreas allograft should be corrected as soon it is suspected. PMID- 10383111 TI - Introduction to structural and functional studies on nerve growth factor. PMID- 10383112 TI - Rita Levi-Montalcini: a brief biographic view of past and present studies on nerve growth factor. PMID- 10383113 TI - Trks: signal transduction and intracellular pathways. AB - The neurotrophin family of growth factors supports survival and differentiation of neurons in the developing vertebrate nervous system by binding activating receptor tyrosine kinases, the Trks. Activation of Trk receptors leads to stimulation of a number of intracellular signaling cascades including, among others, the ras/extracellular regulated kinase (erk) and the phosphatidylinositol 3 kinase (PI 3 kinase) cascades. Over the past several years, work in several neurotrophin responsive systems has begun to identify the role each of these signaling cascades plays in the cellular response to neurotrophins. It now appears that neurotrophins, in particular nerve growth factor (NGF), mediate their multiple effects through a number of distinct intracellular signaling cascades. In this review, we will overview the evidence implicating specific signaling cascades in aspects of the cellular response to the neurotrophins, specifically in response to activation of TrkA by NGF. PMID- 10383114 TI - p75 neurotrophin receptor as a modulator of survival and death decisions. AB - The p75 receptor is the founding member of the TNF receptor superfamily. Members in this receptor family share a common cysteine motif repeated two to six times that serves as the ligand binding domain. In addition, several members contain a cytoplasmic region designated the death domain. The neurotrophins NGF, BDNF, NT 3, and NT-4 each bind to the p75 receptor and also more selectively to members of the Trk family of receptor tyrosine kinases. Although the biological functions of p75 have been elusive, recent experimental evidence supports an involvement of this receptor in apoptosis. This presents a counter-intuitive function for neurotrophins, which are normally required for the survival of neurons during development. The life-and-death decisions by neurotrophins appear to be governed by the level of expression and signaling activities of the p75 and Trk tyrosine kinase receptors and their downstream effector molecules. The generation of the correct number of cells in the nervous system is a highly controlled and coordinated process that is the consequence of cell proliferation and cell death decisions. The appropriate number of neuronal and glial cells formed during development guarantees the establishment of proper innervation and functional synaptic connections. One common mechanism to account for the number of viable cells is the ability to form ligand-receptor complexes that promote cell survival under conditions of limiting concentrations of trophic factors. Another diametrically opposed mechanism is to produce ligand-receptor interactions that can activate programmed cell death directly. PMID- 10383115 TI - Classical and novel directions in neurotrophin transport and research: anterograde transport of brain-derived neurotrophic factor by sensory neurons. AB - After the discovery of nerve growth factor, a classic model of neurotrophin action was developed. In this model, nerve endings compete for limited quantities of neurotrophic factors produced in neuronal target tissues. Neurotrophins are bound with high-affinity receptors expressed on the neuronal membrane and then endocytosed and retrogradely transported back to the cell body of responsive neurons. This classic model of target derived trophic support has been utilized to explain a wide range of trophic actions including effects on neuronal survival, terminal branching, and protein expression. However, a number of recent findings in the field of neurotrophin research cannot be explained using the classic model. In the peripheral nervous system (PNS), sensory neurons have been shown to contain mRNA for a member of the neurotrophin family, brain-derived neurotrophic factor (BDNF). Sensory neurons do not receive synaptic input so neurotrophin production by these cells does not fit into the classic target derived model. In contrast to target derived trophic support, BDNF produced by sensory neurons provides local autocrine and paracrine neurotrophic support in vitro. Furthermore, in vivo, sensory neurons transport BDNF in the anterograde direction away from the cell body, and opposite to the retrograde direction utilized in the classic model. Thus, out of necessity, a new direction for neurotrophin research has developed to study the production and anterograde transport of neurotrophins. The importance of this new mode of neurotrophin action in the PNS is indicated by results that implicate it in the response to pain, inflammation, and nerve injury. PMID- 10383116 TI - Neurotrophin actions during the development of the peripheral nervous system. AB - Neurotrophins are important regulators of the development and maintenance of the vertebrate nervous system. Besides their well-established role in promoting neuronal survival during development, in vitro data suggest that they can regulate proliferation, survival, and differentiation of precursor cells. Analysis of the developing peripheral nervous system in mouse strains carrying mutations in genes encoding the neurotrophins and their receptors indicate, however, that lack of neurotrophin signalling results in specific neuronal deficits that are primarily due to neuronal death. Many of these deficits occur before final target encounter. PMID- 10383117 TI - Neurotrophins and other growth factors in the generation of retinal neurons. AB - The generation of neurons in the vertebrate retina, as in other areas of the developing nervous system, largely depends on extracellular signals. Of the known signaling molecules, neurotrophins play decisive, defined, and distinct roles. The three neurotrophins identified in the chick, namely, neurotrophin-3 (NT-3), brain-derived neurotrophic factor (BDNF), and nerve growth factor (NGF), are expressed in either the pigment epithelium (NT-3 and BDNF) or in the neural retina (NGF) at the onset of neuron birth. In addition, trkC and trkB, receptors for NT-3 and BDNF, respectively, together with p75, the low-affinity neurotrophin receptor, are expressed in the retina at the same developmental period. The role of these three neurotrophins in the differentiation of neurons in the chick retina has been elucidated by a combination of in vitro and in vivo experiments. Thus, NT-3 promotes the conversion of neuroepithelial cells into neurons, whereas BDNF and NGF control the programmed cell death (apoptosis) that affects early postmitotic neuroblasts. BDNF, acting via its trkB receptor, is a survival factor for these cells, whereas NGF, binding to p75 receptor, acts as a killing factor, thereby controlling the provisional number of newly generated neurons. PMID- 10383118 TI - Neurotrophins, nociceptors, and pain. AB - It is now well established that neurotrophins play a crucial role in the development of the nervous system. However, there is increasing evidence that the function of neurotrophins persists throughout adulthood. The broad scope of neurotrophin action is well documented in the case of nerve growth factor (NGF) and its effect on nociceptors and nociception. Here, we review the evidence for these multiple roles for NGF. Two manipulations influencing NGF levels are discussed in detail. The first involves the use of transgenic mice that overexpress or underexpress neurotrophins. A second strategy involves administration of NGF or its antibody in vivo to increase or decrease its level. During prenatal development, NGF is required for survival of nociceptors. In the early postnatal period, NGF is required for expression of the appropriate nociceptor phenotype. In adults, NGF acts as an important intermediate in inflammatory pain, contributing to both peripheral and central sensitization. The sensitization of peripheral nociceptors can be very rapid and can involve non neural cells such as mast cells, neutrophils, fibroblasts, and macrophages. Recent evidence indicates that other neurotrophins also play key supporting roles in the development of nociceptors (e.g., NT-3) and in inflammatory pain (e.g., BDNF, NT-4/5). Furthermore, molecules from other superfamilies (e.g., GDNF) also are required to assure survival of certain classes of nociceptors. The diverse effects of neurotrophins on nociceptive processing emphasize their broad importance in the development and function of the nervous system. PMID- 10383119 TI - Are there differences between the secretion characteristics of NGF and BDNF? Implications for the modulatory role of neurotrophins in activity-dependent neuronal plasticity. AB - In previous experiments the activity-dependent secretion of nerve growth factor (NGF) from native hippocampal slices and from NGF-cDNA transfected hippocampal neurons showed unusual characteristics [Blochl and Thoenen (1995) Eur J Neurosci 7:1220-1228; (1996) Mol Cell Neurosci 7:173-190]. In both hippocampal slices and cultured hippocampal neurons the activity-dependent secretion proved to be independent of extracellular calcium, but dependent on the release of calcium from intracellular stores. Under different experimental conditions, Goodman et al. [(1996) Mol Cell Neurosci 7:222-238] reported that the high potassium mediated secretion of brain-derived neurotrophic factor (BDNF) from hippocampal cultures was dependent on extracellular calcium. Mowla et al. [(1997) Proc 27th Annu Meet Soc Neurosci New Orleans 875.10] reported on even further-reaching differences between NGF and BDNF secretion, namely, that in hippocampal neurons and in pituitary cell lines NGF was secreted exclusively according to the constitutive pathway, whereas BDNF was exclusively sorted according to the activity-dependent regulated pathway. In view of the crucial importance of such potential differences between the processing, sorting, and secretory mechanisms of different neurotrophins for their modulatory roles in activity-dependent neuronal plasticity, a thorough analysis under comparable experimental conditions was mandatory. We demonstrate that in native hippocampal slices and adenoviral transduced hippocampal neurons there are no differences between NGF and BDNF with respect to the subcellular distribution and mechanism of secretion; that the activity-dependent secretion of both NGF and BDNF is dependent on intact intracellular calcium stores; and that the differences between our own observations and those of Goodman et al. (ibid.) regarding the dependence on extracellular calcium do not reflect differences between NGF and BDNF sorting and secretion, but reflect the differing experimental conditions used. PMID- 10383120 TI - Neurotrophin-mediated potentiation of neuronal injury. AB - The neurotrophins are a diverse family of peptides which activate specific tyrosine kinase-linked receptors. Over the past five decades, since the pioneering work of Levi-Montalcini and colleagues, the critical role that neurotrophins play in shaping the developing nervous system has become increasingly established. These molecules, which include the nerve growth factor (NGF)-related peptides, NGF, brain-derived neurotrophic factor (BDNF), NT-4/5 and NT-3, promote differentiation and survival in the developing nervous system, and to a lesser extent in the adult nervous system. As survival-promoting molecules, neurotrophins have been studied as potential neuroprotective agents, and have shown beneficial effects in many model systems. However, a surprising "dark side" to neurotrophin behavior has emerged from some of these studies implying that, under certain pathological conditions, neurotrophins may exacerbate, rather than alleviate, injury. How neurotrophins cause these deleterious consequences is a question which is only beginning to be answered, but initial work supports altered free radical handling or modification of glutamate receptor expression as possible mechanisms underlying these effects. This review will focus on evidence suggesting that neurotrophins may enhance injury under certain circumstances and on the mechanisms behind these injury-promoting aspects. PMID- 10383121 TI - Nerve growth factor: a neurotrophin with activity on cells of the immune system. AB - Numerous studies published in the last two decades provide evidence that nerve growth factor (NGF), a polypeptide originally discovered because of its neurotrophic activity, acts on a variety of cells of the immune system, including mast cells, eosinophils, and B and T lymphocytes. NGF has been shown to increase during inflammatory responses, autoimmune disorders, parasitic infections, and allergic diseases. Moreover, stress, which is characterized also by activation of a variety of immune cells, causes a significant increase in basal plasma NGF levels. Recently published studies reveal that hematopoietic progenitor cells seem to be able to produce and/or respond to NGF. We report these data and discuss the hypothesis of the possible implication of NGF on the functional activities of immune cells. PMID- 10383122 TI - Other neurotrophic factors: glial cell line-derived neurotrophic factor (GDNF). AB - Glial cell line-derived neurotrophic factor (GDNF) was first discovered as a potent survival factor for midbrain dopaminergic neurons and was then shown to rescue these neurons in animal models of Parkinson's disease. GDNF is a more potent survival factor for dopaminergic neurons and the noradrenergic neurons of the locus coeruleus than other neurotrophic factors, and an almost 100 times more efficient survival factor for spinal motor neurons than the neurotrophins. The members of the GDNF family, GDNF, neurturin (NTN), persephin (PSP), and artemin (ART), have seven conserved cysteine residues with similar spacing, making them distant members of the transforming growth factor-beta (TGF-beta) superfamily. Like the members of the neurotrophin family, the GDNF-like growth factors belong structurally to the cysteine knot proteins. Like neurotrophins, GDNF family proteins are responsible for the development and maintenance of various sets of sensory and sympathetic neurons but, in addition, GDNF and NTN are also responsible for the development and survival of the enteric neurons, and NTN for parasympathetic neurons. All neurotrophins bind to the p75 low-affinity receptor, but their ligand specificity is determined by trk receptor tyrosine kinases. GDNF, NTN, PSP, and ART mediate their signals via a common receptor tyrosine kinase, Ret, but their ligand specificity is determined by a novel class of glycosylphosphatidylinositol (GPI)-anchored proteins called the GDNF family receptor alpha (GFR alpha). GDNF binds preferentially to GFR alpha1, NTN GFR alpha2, ART GRF alpha3, and PSP GFR alpha4 as a co-receptor to activate Ret. GFR alpha4 has until now been described only from chicken. Although the GDNF family members signal mainly via Ret receptor tyrosine kinase, there is recent evidence that they can also mediate their signals via GFR alpha receptors independently of Ret. The GDNF family of growth factors, unlike neurotrophins, has a well-defined function outside the nervous system. Recent transgenic and organ culture experiments have clearly demonstrated that GDNF is a mesenchyme-derived signaling molecule for the promotion of ureteric branching in kidney development. NTN, ART, and PSP are also expressed in the developing kidney, and NTN and PSP induce ureteric branching in vitro, but their true in vivo role in kidney morphogenesis is still unclear. PMID- 10383123 TI - Comparison of eight histomorphometric methods for measuring trabecular bone architecture by image analysis on histological sections. AB - Osteoporosis is defined as a disease characterized by low bone mass and microarchitectural deterioration of trabecular bone leading to enhanced bone fragility. Various histomorphometric methods have been described to measure bone architecture on histological sections. However, not all of the methods are strictly equivalent and some of them appear able to detect differences earlier in the course of the disease. We have compared 8 histomorphometric methods known to characterize the architecture of trabecular bone in 154 male osteoporotic patients. Measurements were done on transiliac bone biopsies: Trabecular number, thickness, and separation (Tb.N, Tb.Th, Tb.Sp); Trabecular Bone Pattern Factor (TBPf); Euler-Poincare's number (E); Interconnectivity Index (ICI); strut analysis of the trabecular network with the ratio of nodes/free-end (N/F); star volume of the bone marrow (V*m.space) and trabeculae (V*Tb) and the Kolmogorov fractal dimension of the trabecular boundaries (D). Relationships between the various architectural parameters were studied by hierarchical cluster analysis. Linear, hyperbolic, and exponential correlations were found between trabecular bone volume (BV/TV) and architectural parameters. Cluster analysis demonstrates the link between these architectural parameters. ICI, E, and TBPf, which reflect the amount of open/closed marrow cavities clustered together and appeared related to Tb.Sp, V*m.space which are indicators of the mean size of marrow cavities. Tb.Th, V*Tb and N/F flocked together as they reflect the trabecular size. Tb.N and D segregated together and seemed to best describe the trabecular network complexity. These histomorphometric techniques are correlated but correlations may be linear or nonlinear. Several histomorphometric techniques need to be used in parallel to appreciate the pathophysiological mechanisms of osteoporotic states. PMID- 10383124 TI - Sensilla on the antenna and ovipositor of the parasitic wasps trichogramma galloi Zucchi and T. pretiosum Riley (Hym., Trichogrammatidae). AB - The understanding of the stimuli perceived by these parasitoids to accept and exploit a potential host may support the development of artificial hosts for their in vitro rearing. The most common structures involved in the host selection process in parasitoids are the antenna and the ovipositor. Sensilla present on these structures are involved in host recognition and acceptance, and the identification of which kind of stimulus (physical or chemical) is perceived by the sensilla could provide data about the host characteristics that elicit the parasitism behavior. The antenna and ovipositor of the wasps Trichogramma galloi and T. pretiosum were studied to identify their sensillar structure and their possible functions in the host selection process, as a support to the development of an artificial host for the in vitro rearing of these parasitoids. Seven sensilla and one setiform structure were present on the antenna of both parasitoid species, with a mixed, chemo-, or mechanoreception function. The ovipositor has four different sensilla and their role in the host evaluation is suggested. PMID- 10383125 TI - Ultrastructure of the olfactory epithelium in intact, axotomized, and bulbectomized goldfish, Carassius auratus. AB - The ultrastructure of the olfactory epithelium in intact, axotomized, and bulbectomized goldfish was studied by scanning and transmission electron microscopy. A total of 58 adult goldfish of various survival times were examined to determine whether the different types of surgery--either olfactory nerve transection or bulbectomy--yielded differences in the extent or time course of cellular degeneration and renewal. Control animals were also examined in detail to elucidate previous controversial findings concerning the types of olfactory receptor neurons present in goldfish. We found that the intact olfactory epithelium of unoperated control goldfish contains the previously observed ciliated and microvillous receptor neurons, and the crypt cell, a cell type not yet seen in the goldfish but recently reported in other species of teleosts. Following either olfactory nerve transection or bulbectomy, the olfactory receptor neurons showed similar signs of degeneration and subsequent cell death, but, surprisingly, the thickness of the olfactory epithelium did not change significantly with either treatment. The time course of receptor cell renewal was different in axotomized and bulbectomized goldfish. In axotomized goldfish, the amount of receptor cells decreased continuously until 8-13 days after surgery, followed by rapid cell renewal. For bulbectomized goldfish, cell replacement began almost immediately after surgery, with degeneration and cell renewal occurring simultaneously. Six weeks after bulbectomy, cell death and cell proliferation reached a "steady state," and the epithelia did not further improve. PMID- 10383126 TI - Quantitation of MYC gene expression in sporadic breast tumors with a real-time reverse transcription-PCR assay. AB - MYC gene overexpression was identified recently as a downstream step at the end of the Wnt/APC/beta-catenin pathway dysregulation observed in colorectal cancer (T-C. He et al., Science (Washington DC), 281: 1509-1512, 1998). It thus appears that an excess of c-myc protein is a primary cause of numerous cancers. In breast cancer, MYC has been studied mostly at the DNA level because of the poor quality of available antibodies against the protein product. The renewed interest in MYC calls for a sensitive and accurate method for analyzing MYC overexpression in breast tumors. We have developed a real-time quantitative reverse transcription PCR assay based on TaqMan fluorescence methodology to quantify the MYC mRNA copy number. We validated the method on a large series of breast tumors. MYC gene overexpression was observed in 29 of 134 (22%) breast tumor RNAs, ranging from 3.2 to 19 times the level in normal breast tissues. These data imply that dysregulated MYC gene expression is potentially involved in the pathogenesis of breast cancer, especially by favoring local cell proliferation. We also found that MYC gene overexpression was rarely due to an increased MYC gene copy number in breast cancer. This new, simple, rapid, and semiautomated method will be useful for screening cancer patients for MYC overexpression and will prove a powerful tool in large, randomized, prospective, cooperative group trials and in the MYC-based therapy project. PMID- 10383127 TI - Inhibitors of histone deacetylase relieve ETO-mediated repression and induce differentiation of AML1-ETO leukemia cells. AB - The (8;21) translocation, found in 12% of acute myeloid leukemia (AML), creates the chimeric fusion product, AML1-ETO. Previously, we demonstrated that the ETO moiety recruits a transcription repression complex that includes the histone deacetylase (HDAC1) enzyme. Here, we used inhibitors of HDAC1 to study the pathophysiology of AML1-ETO. Both the potent inhibitor, trichostatin (TSA), and the well-known but less specific inhibitor, phenylbutyrate (PB), could partially reverse ETO-mediated transcriptional repression. PB was also able to induce partial differentiation of the AML1-ETO cell line, Kasumi-1. With the intention of developing a clinically useful protocol, we combined PB with a number of other agents that induced differentiation and apoptosis of Kasumi-1 cells. In summary, transcriptional repression mediated by AML1-ETO appears to play a mechanistic role in the t(8;21) AML, and relief of repression using agents such as PB (alone or in combination) may prove to be therapeutically useful. PMID- 10383128 TI - Molecular cloning and functional analysis of the mouse homologue of the KILLER/DR5 tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) death receptor. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and its receptors are members of the tumor necrosis factor superfamily. TRAIL selectively kills cancer cells but not normal cells. We report here the cloning of the mouse homologue of the TRAIL receptor KILLER/DR5 (MK). The cDNA of MK is 1146 bp in length and encodes a protein of 381 amino acids. MK contains an extracellular cysteine-rich domain, a transmembrane domain, and a cytoplasmic death-domain characteristic of Fas, tumor necrosis factor, and human TRAIL receptors. MK is highly homologous and binds TRAIL with similar affinity as human DR4 and KILLER/DR5. MK induces apoptosis in mouse and human cells and inhibits colony growth of NIH3T3 cells. Expression of MK is p53-dependent and up-regulated by tumor suppressor p53 and by DNA damaging agents in mouse cells undergoing apoptosis. This is the first report describing a mouse TRAIL receptor gene and also demonstrating that the p53-dependent regulation of KILLER/DR5-mediated apoptosis is conserved between human and mouse. PMID- 10383129 TI - Recurrent involvement of 2p23 in inflammatory myofibroblastic tumors. AB - Inflammatory myofibroblastic tumor (IMT) is a relatively rare soft tissue tumor. The reactive versus neoplastic pathogenesis of this tumor is unresolved. We found clonal chromosome aberrations involving 2p23 upon metaphase analysis of two IMTs. Fluorescence in situ hybridization with a probe flanking the ALK gene at 2p23 demonstrated rearrangement of the probe in both of these cases and in a third case, and immunohistochemistry revealed ALK expression in all three cases. 2p22 24 involvement has been reported previously in four additional cases of IMT. We suggest that chromosomal rearrangements involving 2p23 near or within ALK are recurrent alterations in IMT and that ALK may have a novel role outside its previously recognized realm of lymphoid neoplasms. PMID- 10383130 TI - The transcriptional activities of p53 and its homologue p51/p63: similarities and differences. AB - p51/p63 is a novel p53 homologue that has been shown to act as a transcriptional activator through the p53-binding sequence of the p21/WAF1 promoter and to induce apoptosis when it is expressed transiently in a human tumor cell line. We developed transcription assay systems for these two related genes in both Saccharomyces cerevisiae and mammalian cells and used them to investigate the functional similarities and differences of these genes. We found that p51/p63 trans-activated the previously identified p53 target genes, but the degree of the transactivation by p51/p63 differed from that by p53. These results suggest that the cellular signal on p51/p63 cross-talks partially but not completely with that of the p53 pathway. PMID- 10383131 TI - Human prostate cancer expresses the low affinity insulin-like growth factor binding protein IGFBP-rP1. AB - Many of the alterations in the insulin-like growth factor (IGF) axis in prostatic disease have been associated with changes in the insulin-like growth factor binding proteins (IGFBPs), a multigene family of proteins that are thought to mediate the action of IGFs on target tissues. IGFBP-related protein 1 (rP1), also known as IGFBP-7 or mac25, is a recently described member of the IGFBP family, the biological function of which has yet to be completely ascertained. In this study, we analyzed the localization of IGFBP-rP1 in prostate cancer and benign prostate tissues using immunohistochemistry and a polyclonal antibody, T1A12, that is specific for IGFBP-rP1. The most intense staining was observed in nerves, whereas smooth muscle cells in the prostate stained weakly. Lymphocytes were always negative. When normal prostatic secretory epithelium was present, staining was usually absent. The lining secretory epithelium stained positively in 0 of 12 (0%) cases of benign prostatic hyperplasia, 57 of 63 (90.5%) primary adenocarcinomas, and 7 of 7 (100%) prostate cancer metastases. Prostatic intraepithelial neoplasia showed a similar pattern of staining to that observed for the invasive tumors. Analysis of Northern blots showed that none of the prostate cancer cell lines (LNCaP, C4, C4-2, C4-2B4, 9069E3, DU145, and PC3) expressed IGFBP-rP1 mRNA. This lack of expression was confirmed by immunohistochemistry of s.c.-generated tumor xenografts of LNCaP and C4-2 and by immunoblot on serum-free-conditioned media from all prostatic cell lines. In contrast to these results, tumor xenografts generated by direct intraosseous injection of LNCaP or C4-2 to bone marrow space resulted in tumors that stained positively for IGFBP-rP1. Our results show that IGFBP-rP1 is expressed in both in situ and invasive prostate neoplasms, but not typically in normal secretory or BPH epithelium; furthermore, the expression of IGFBP-rP1 can be induced in human prostate cancer cell lines in vivo on interaction with an appropriate host environment. PMID- 10383132 TI - Elevated and biallelic expression of p73 is associated withprogression of human bladder cancer. AB - p73, a member of the p53 family at 1p36.3, has been demonstrated to be expressed monoallelically and induce apoptosis or G1 arrest of the cell cycle. To explore the candidacy of p73 as a suppressor in bladder tumorigenesis, we examined expression level, allelic origin, and mutation of p73 mRNA in 45 primary bladder carcinomas. Quantitative PCR analysis showed no allelic loss of the gene but showed various levels of mRNA expression in both carcinoma and noncancerous tissues. Elevated expression of p73 was frequently observed in carcinoma tissues [18 (40.0%) of 45] and showed a strong correlation with tumor stage or grade. Allotyping analysis using a StyI polymorphism detected biallelic expression in 12 (52.2%) of 23 heterozygous carcinomas but none in 4 noncancerous tissues. Tumor specific biallelic expression was also identified from one matched set. In addition, 8 (66.7%) of these 12 expressed high levels of p73 mRNA, whereas only 2 (18.2%) of 11 monoallelic expressors showed high expression, which suggests that the increased expression of p73 might be caused by the transcriptional activation of a silent allele in carcinomas. Single-strand conformational polymorphism analysis of the entire coding region of p73 revealed no mutation, whereas 12 (26.7%) of the same set showed p53 alterations. No relationship between expression of p73 and p53 mutation or expression of p21Waf1 or MDM2 was identified. Taken together, our data argue that p73 does not play a role as a tumor suppressor in bladder carcinogenesis and suggest that the activation of a silent allele may contribute to the progression of bladder tumors. PMID- 10383133 TI - Prognostic significance of polo-like kinase (PLK) expression in squamous cell carcinomas of the head and neck. AB - Previously, we demonstrated that the mammalian polo-like kinase (PLK), which participates in the regulation of the cell cycle, is a novel marker of cellular proliferation. Because current prognostic tools for the evaluation of patients with head and neck squamous cell cancer (HNSCC) need to be improved, we analyzed 89 patients and found elevated PLK expression in most tumors. Nodal stage as a crucial prognostic factor in HNSCC also correlated to PLK transcript levels (P = 0.0043). A Kaplan-Meier analysis showed that HNSCC patients with moderate versus high PLK expression survived significantly longer (5-year survival rates, 43% versus 12%; P = 0.0047). Interestingly, a combination of nodal stage and PLK expression contributed to discriminate patients with a better prognosis in the pN(0/1) and pN(2/3) groups, which could improve the definition of a suitable therapy. PMID- 10383134 TI - Eponemycin exerts its antitumor effect through the inhibition of proteasome function. AB - Cell cycle progression requires the proteasome-mediated degradation of key regulatory proteins such as cyclins, cyclin-dependent kinase inhibitors, and anaphase-inhibitory proteins. Given the central role of the proteasome in the destruction of these proteins, proteasome inhibition has been proposed as a possible cancer therapy. We report here that dihydroeponemycin, an analogue of the antitumor and antiangiogenic natural product eponemycin, selectively targets the 20S proteasome. Dihydroeponemycin covalently modifies a subset of catalytic proteasomal subunits, binding preferentially to the IFN-gamma-inducible subunits LMP2 and LMP7. Moreover, the three major peptidolytic activities of the proteasome are inhibited by dihydroeponemycin at different rates. In addition, dihydroeponemycin-mediated proteasome inhibition induces a spindle-like cellular morphological change and apoptosis. These results validate the proteasome as a target for antitumor pharmacological intervention and are relevant for the design of novel chemotherapeutic strategies. PMID- 10383135 TI - Adoptive transfer of immature dendritic cells with autologous or allogeneic tumor cells generates systemic antitumor immunity. AB - Dendritic cells (DCs) are potent antigen-presenting cells that are capable of priming systemic antitumor immune responses in animal tumor models. However, many of the model tumor systems tested need definition of the specific tumor antigens involved. To use DCs in situations that are more relevant to the majority of human cancers, where the antigens are unknown, we have tested the adoptive transfer of immature DCs in mouse colorectal and melanoma models of varying immunogenicity but with undefined antigens. When DCs admixed with a syngeneic primary tumor inoculum were seeded s.c., the growth of the primary tumor was unchanged; however, if the primary tumor was then surgically excised and the animal was rechallenged with the same tumor, significant protection (75%) was generated when DCs were present in the original primary inoculum of a moderately immunogenic colorectal model (CMT93tk). This effect was not observed when a nonimmunogenic melanoma (B16) was tested in an identical protocol. Next, DCs were injected directly into 6-9-mm established tumors; again, protection (55%) was achieved against a secondary tumor challenge following excision of the primary, but only in the CMT93tk model of moderate immunogenicity. To increase the clinical relevance of this approach still further, we tested irradiated allogeneic K1735 melanoma cells mixed with syngeneic DCs as a vaccine against subsequent challenge with the poorly immunogenic syngeneic melanoma B16. The allogeneic vaccine alone was ineffective, but when admixed with DCs, a significant number of animals rejected a subsequent B16 challenge, suggesting that DCs are able to prime an immune response against melanoma antigens shared between K1735 and B16. The generation of systemic antitumor immunity by adoptive transfer of DCs has significant clinical potential because it is technically straightforward and does not require the definition of specific tumor antigens. PMID- 10383136 TI - Identification of a region of frequent loss of heterozygosity at 11q24 in colorectal cancer. AB - Loss of heterozygosity (LOH) at 11q23-qter occurs frequently in ovarian and other cancers, but for colorectal cancer, the evidence is conflicting. Seven polymorphic loci were analyzed between D11S897 and D11S969 in 50 colorectal tumors. Two distinct LOH regions were detected, suggesting possible sites for tumor-suppressor genes involved in colorectal neoplasia: a large centromeric region between D11S897 and D11S925, and a telomeric 4.9-Mb region between D11S912 and D11S969. There was no correlation with clinicopathological features. This analysis describes a region of LOH in the region 11q23.3-24.3 for the first time in colorectal cancer and provides complementary evidence for the ongoing effort to identify the gene(s) involved. PMID- 10383137 TI - Identification of a transactivation activity in the COOH-terminal region of p73 which is impaired in the naturally occurring mutants found in human neuroblastomas. AB - p73 is a recently cloned tumor suppressor gene that is highly homologous to p53, and the products of both possess similar functions in inhibiting cell growth and inducing apoptosis. Interestingly, the COOH-terminal region of p53 displays no significant homology with that of p73. Moreover, p73 has an additional segment at its COOH terminus. Recently, we have found two mutations of p73 with amino acid substitution (P405R and P425L) in primary neuroblastomas. Because the region (amino acid residues 382-491) contains a glutamine- and proline-rich domain, we hypothesized that it has a transactivation function, and the mutations found in tumors result in loss of function. To test it, we used the yeast GAL4 DNA-binding fusion system. Yeast transformants expressing a GAL4-p73(1-112) or a GAL4 p73alpha(380-513) fusion protein were grown in SD medium lacking histidine and tryptophan and exhibited a significant induction of beta-galactosidase activity. Transient transfection experiments revealed that both of fusion proteins could induce the chloramphenicol acetyltransferase activity in mammalian cells, indicating that the COOH-terminal as well as NH2-terminal regions of p73 had significantly high levels of transactivation activity. Furthermore, the former activity was severely impaired in two naturally occurring mutant forms found in neuroblastomas. These suggest that, unlike p53, p73 has two domains with transactivation function, one in the NH2-terminal region and the other in the COOH-terminal region. Loss of function mutation in the latter might be involved in tumorigenesis and/or tumor progression. PMID- 10383138 TI - Activation of mitochondrial Raf-1 is involved in the antiapoptotic effects of Akt. AB - The Akt serine/threonine kinase is required for the survival of many cell types and for transformation of hematopoietic cells by the BCR/ABL oncogenic tyrosine kinase. Analysis of the potential mechanisms whereby Akt promotes survival of hematopoietic cells revealed that it induced the activity of plasma membrane and mitochondrial Raf-1 in a Ras-independent, but PKC-dependent manner. Inhibition of plasma membrane Raf-1-dependent mitogen-activated protein kinase activity had no effect on the enhanced survival of cells expressing Akt. By contrast, suppression of mitochondrial Raf-1 enzymatic activity by expression of a mitochondria targeted Raf-1 dominant-negative mutant rendered Akt-expressing cells susceptible to apoptosis induced by growth factor deprivation and was accompanied by inhibition of BAD, but not mitogen-activated protein kinase, phosphorylation. Together, these data indicate that PKC-dependent activation of Raf-1 plays an important role in Akt-dependent antiapoptotic effects. PMID- 10383139 TI - Molecular cloning and expression of an alternative hKLK3 transcript coding for a variant protein of prostate-specific antigen. AB - We report the molecular cloning of a full-length cDNA corresponding to a 2.1-kb hKLK3 mRNA. This mRNA results from the alternative splicing of intron 4, and its accumulation in prostatic LNCaP cells is stimulated by androgen. The cDNA encodes a prepro-prostate-specific antigen (PSA) variant containing 238 amino acids. The new protein, PSA-related protein 1 (PSA-RP1), differs from PSA at the COOH terminal end and lacks the serine residue that is essential for catalytic activity. Prepro-PSA-RP1 was transiently expressed in COS1 cells fused to the V5 epitope of the paramyxovirus SV5. The recombinant fusion protein was detected in the spent medium by Western blot analysis using anti-V5 and anti-PSA antibodies. This indicates that PSA-RP1 is secreted and has PSA-like antigenic epitopes. A pro-PSA and a pro-PSA-RP1 having a mutated propeptide were overproduced in Escherichia coli fused to glutathione S-transferase. The recombinant PSA and PSA RP1 were matured in vitro and identified by Western blot with molecular masses of 29 (PSA) and 27 (PSA-RP1) kDa. The data indicate that PSA-RP1, not complexed to serine protease inhibitors, could be present in biological fluids, thus contributing to the free PSA-immunoreactive fraction in serum. PMID- 10383140 TI - CYP17 and breast cancer risk: the polymorphism in the 5' flanking area of the gene does not influence binding to Sp-1. AB - The ability of a motif of the CYP17 5' untranslated region, created by a polymorphic T to C substitution, to bind to the human transcription factor Sp-1 was investigated. No binding of any of the polymorphic alleles was observed in electromobility shift assay. No other sequence within +1 to +100 of each of the CYP17 alleles formed complex with the Sp-1 or enhanced binding to the polymorphic CACC box. Genotyping of 510 breast cancer patients and 201 controls revealed no difference in genotype frequencies. Age at onset, tumor grade, lymph node status and distant metastases, stage, and estrogen and progesterone receptor status were not associated with the CYP17 genotype. PMID- 10383141 TI - Implication of p53 in growth arrest and apoptosis induced by the synthetic retinoid CD437 in human lung cancer cells. AB - CD437 is a novel retinoid that can induce apoptosis in a variety of tumor cell types by an unknown mechanism. We found that CD437 up-regulated the expression of p21(WAF1/CIP1), Bax, and Killer/DR5 and induced G1 arrest and rapid apoptosis in three human non-small cell lung carcinoma cell lines with wild-type p53 but not in five cell lines with mutant p53, suggesting a role for p53 in the effects of CD437. Using H460 cells in which wild-type p53 protein was degraded by transfection of the human papillomavirus 16 E6 (HPV-16 E6) gene and H460 cells transfected with a control plasmid only, we found that CD437 increased p53, p21(WAF1/CIP1), Bax, and Killer/DR5 in the control transfectants. In contrast, the constitutive p53 protein level was suppressed, and the ability of CD437 to increase p53 and its downstream genes was compromised in E6 transfectants. In addition, CD437 induced G1 arrest and apoptosis in the control transfectants but not in the E6-transfected cells. These results indicate that p53 plays a role in CD437-induced growth inhibition and apoptosis in human non-small cell lung carcinoma cells. PMID- 10383142 TI - Optimizing syngeneic orthotopic murine bladder cancer (MB49). AB - The syngeneic orthotopic murine bladder cancer model MB49 is hampered by unreliable tumor implantation. We optimized this model by a simple modification of the standard implantation technique in three groups of mice. Fifty thousand (group I), 20,000 (group II), or 10,000 (group III) tumor cells were implanted into cauterized bladders by transurethral instillation, and dwell time was prolonged to 3 h. Tumor take, survival, and bladder weights were determined as outcome variables. To verify whether this modification maintained its sensitivity to topical immunotherapy, an initial tumor load of 100,000 MB49 cells was given, and mice were treated intravesically with Bacillus Calmette-Guerin or phosphate buffered saline. The prolonged dwell time of tumor cells resulted in take rates of 100% in all three groups. Survival and bladder weights were significantly correlated with the number of instilled cells. Even with the highest tumor load, Bacillus Calmette-Guerin therapy improved survival and reduced bladder weights significantly, as compared to PBS. Thus, the modified model is highly reliable and maintains its susceptibility to topical immunotherapy. PMID- 10383143 TI - Dead or dying: necrosis versus apoptosis in caspase-deficient human renal cell carcinoma. AB - The antitumor effect of immuno- and chemotherapeutic agents is executed through stimulation of apoptotic programs in susceptible cells. Apoptosis induced in tumor cells requires activation of members of the caspase family of proteases. Deficient expression or activation of caspases may account in part for the failure of many current anticancer therapies. However, recent studies suggest that cell death can proceed in the absence of caspases. We investigated the susceptibility of human renal cell carcinoma (RCC) lines to two distinct modes of cell death, apoptosis and necrosis. RCC lines displayed almost complete resistance to apoptosis in response to the intracellular zinc chelator, N,N,N'N' tetrakis (2-pyridylmethyl) ethylenediamine (TPEN), which instead induced dramatic accumulation of nonapoptotic necrotic cells. Conversely, TPEN was a potent inducer of apoptosis in caspase-competent normal kidney cells (NK-72) and Jurkat T lymphocytes. Resistance to apoptosis in RCC lines correlated with almost complete loss of caspase-3 expression and variable down-regulation of caspase-7, caspase-8, and caspase-10. These data may explain the resistance of RCC to drugs inducing apoptosis and have important consequences for further attempts to manipulate tumor cell death. PMID- 10383144 TI - Transcriptional up-regulation of paxillin expression by heregulin in human breast cancer cells. AB - Activation of heregulin (HRG) signaling has been implicated in the development of aggressive phenotype in breast cancer cells. The mechanisms through which HRG regulates the progression of breast cancer cells to a more invasive or motile phenotype are currently unknown. Because the process of cell migration must involve dynamic changes in the formation of new focal adhesions at the leading edge and dissolution of preexisting focal points, we explored the potential HRG regulation of paxillin, a major component of focal adhesion. Here, we report that HRG stimulation of noninvasive breast cancer MCF-7 cells resulted in the up regulation of paxillin mRNA and protein. The observed HRG stimulation of paxillin mRNA expression was completely blocked by actinomycin D (a transcriptional inhibitor) as well as by cycloheximide (a protein synthesis inhibitor), suggesting the involvement of an inducible protein factor(s) and transcriptional regulation of paxillin mRNA by HRG. Extension of these observations to other HRG responsive human cell lines also demonstrated that HRG has a significant capacity to up-regulate the paxillin expression. Furthermore, the levels of paxillin expression were closely linked with the coexpression of human epidermal growth factor receptor 2 (HER2)/HER3 receptors in breast cancer cell lines and in grade III human breast tumors. This study is the first demonstration of regulation of paxillin expression by a polypeptide growth factor, and it suggests a potential role for paxillin in the HER2 pathway in breast cancer. PMID- 10383145 TI - Antagonism of p53-dependent apoptosis by mitogen signals. AB - p53-mediated apoptosis is antagonized by growth factor stimulation. Here, we show that p53-dependent cell death induced by DNA damage was effectively prevented by mitogen activation. The levels of Bcl-2, Bcl-xL, and Bax were not altered by cisplatin treatment and mitogen rescue. Instead, the protection against p53 regulated apoptosis was mediated by at least three distinct signaling pathways. Either phosphatidylinositol (PI) 3-kinase or mitogen-activated protein kinase kinase (MEK) antagonized p53-induced apoptosis, and an additive preventive effect was observed when both kinases were activated. However, the combination of PI 3 kinase and MEK was not sufficient to completely prevent apoptosis induced by DNA damage. Mitogen activation further suppressed cisplatin-induced p53 expression, and the inhibition was mainly dependent on the Ca2+ pathway. Our results demonstrate that effective antagonism of p53-dependent apoptosis by mitogenic activation requires the presence of multiple signal pathways, including PI 3 kinase, MEK, and Ca2+. PMID- 10383146 TI - Intercalation into DNA is not required for inhibition of topoisomerase I by indolocarbazole antitumor agents. AB - The DNA-intercalating antitumor drug NB-506 is a potent topoisomerase poison currently undergoing phase I/II clinical trials. It contains a planar indolocarbazole chromophore substituted with a glucose residue. Up until now, it was thought that intercalation of the drug into DNA was essential for the stabilization of topoisomerase I-DNA covalent complexes. But, in the present study, we show that a regio-isomeric form of NB-506 has lost its capacity to intercalate into DNA, but remains an extremely potent topoisomerase I poison. The new analogue contains two hydroxyl groups at positions 2,10 instead of positions 1,11 in NB-506. The relocation of the two OH groups reduces considerably the strength of binding to DNA and prevents the drug from intercalating into the DNA double helix. However, the topoisomerase I inhibition capacity of the new analogue remains very high. The two drug isomers are equally potent at maintaining the integrity of the topoisomerase I-DNA covalent complexes, but stimulate cleavage at different sites on DNA. NB-506 stabilizes topoisomerase I preferentially at sites having a pyrimidine (T or C) and a G on the 5' and 3' sides of the cleaved bond, respectively. The 2,10-isomer induces topoisomerase I mediated cleavage only at TG sites and, thus, behaves exactly as the reference topoisomerase I poison camptothecin. Finally, cytotoxicity measurements performed with a panel of murine and human cancer cell lines reveal that the newly designed drug is considerably (up to 100-fold) more toxic to tumor cells than the parent drug NB-506. We conclude that the DNA-binding and topoisomerase I poisoning activities of NB-506 can be viewed as two separate mechanisms. PMID- 10383147 TI - Blocking transforming growth factor beta signaling in transgenic epidermis accelerates chemical carcinogenesis: a mechanism associated with increased angiogenesis. AB - Mutations in the transforming growth factor beta type II receptor (TGF-betaRII) have been identified in human cancers, which suggests a causal role for the loss of TGF-betaRII in cancer development. To directly test this in vivo, we have generated transgenic mice expressing a dominant negative TGF-betaRII (delta betaRII) in the epidermis, using a truncated mouse loricrin promoter (ML). ML.delta betaRII transgenic mice exhibited a thickened skin due to epidermal hyperproliferation. When these mice were subjected to a standard two-stage chemical carcinogenesis protocol, they exhibited an increased sensitivity, with an earlier appearance and a 2-fold greater number of papillomas than control mice. In addition, papillomas in control mice regressed after termination of 12-O tetradecanoylphorbol-13-acetate (TPA) treatment; whereas ML.delta betaRII papillomas progressed to carcinomas. Furthermore, TPA promotion alone induced papilloma formation in ML.delta betaRII mice, which suggests an initiating role for delta betaRII in skin carcinogenesis. ML.delta betaRII tumors also exhibited increased neovascularization and progressed to metastases, although the primary tumors were still classified as carcinoma in situ or well-differentiated carcinomas. Increased expression of vascular endothelial growth factor, an angiogenesis factor, and decreased expression of thrombospondin-1, an angiogenesis inhibitor, were also observed in ML.delta betaRII tumors. The increased angiogenesis correlated with elevated endogenous TGF-beta1 in ML.delta betaRII tumors. These data provide in vivo evidence that inactivation of TGF betaRII accelerates skin carcinogenesis at both earlier and later stages, and increased angiogenesis is one of the important mechanisms of accelerated tumor growth and metastasis. PMID- 10383148 TI - NG2 proteoglycan-binding peptides target tumor neovasculature. AB - NG2 is the rat homologue of the human melanoma proteoglycan, also known as the high molecular weight melanoma-associated antigen. This developmentally regulated membrane-spanning chondroitin sulfate proteoglycan is expressed primarily by glial, muscle, and cartilage progenitor cells. Upon maturation, these cell types down-regulate NG2 expression. In adult animals, the expression of NG2 is restricted to tumor cells and angiogenic tumor vasculature, making this proteoglycan a potential target for directing therapeutic agents to relevant sites of action. To this end, we have identified specific NG2-binding peptides by screening a phage-displayed random peptide library on purified NG2. Several rounds of biopanning on NG2 resulted in the specific enrichment of two phage displayed decapeptides, TAASGVRSMH and LTLRWVGLMS. The binding of these phages to NG2 was inhibitable both by soluble NG2 and by glutathione S-transferase (GST) fusion proteins containing the cognate peptide sequences. In addition, direct binding between GST-TAASGVRSMH and GST-LTLRWVGLMS fusion proteins and NG2 was demonstrated in solid-phase binding assays. Interestingly, these NG2-binding fusion proteins cross-inhibited each other's binding to NG2, suggesting that the two sequences bind to the same or overlapping sites on the proteoglycan. Upon injection into tumor-bearing mice, NG2-binding phages specifically homed to tumor vasculature in wild-type mice but did not localize to the tumor vasculature in NG2 knockout mice. The in vivo targeting capability of these sequences suggests that they can be used for tumor targeting. PMID- 10383149 TI - Effects of chronic low-dose ultraviolet B radiation on DNA damage and repair in mouse skin. AB - Chronic exposure to sunlight causes skin cancer in humans, yet little is known about how habitual exposure to low doses of ultraviolet B radiation (UVB) affects DNA damage in the skin. We treated Skh-1 hairless mice with daily doses of suberythemal UVB for 40 days and analyzed the amount and distribution of DNA photodamage using RIAs and immunofluorescence micrography. We found that DNA damage accumulated in mouse skin as a result of chronic irradiation and that this damage persisted in the dermis and epidermis for several weeks after the chronic treatment was terminated. Although the persistent damage was evenly distributed throughout the dermis, it remained in the epidermis as a small number of heavily damaged cells at the dermal-epidermal boundary. Rates of DNA damage induction and repair were determined at different times over the course of chronic treatment in response to a higher challenge dose of UVB light. The amount of damage induced by the challenge dose increased in response to chronic exposure, and excision repair of cyclobutane pyrimidine dimers and pyrimidine(6-4)pyrimidone dimers was significantly reduced. The sensitization of mouse epidermal DNA to photoproduct induction, the reduction in excision repair, and the accumulation of nonrepairable DNA damage in the dermis and epidermis suggest that chronic low dose exposure to sunlight may significantly enhance the predisposition of mammalian skin to sunlight-induced carcinogenesis. PMID- 10383150 TI - Expression of cyclin A in soft tissue sarcomas correlates with tumor aggressiveness. AB - Cyclins and cyclin-dependent kinases regulate the cell cycle. Cyclin A has a dual role in cell proliferation. It is essential in the S phase for DNA replication, and it is also involved in G2-M-phase transition, signifying actively dividing cells. The expression of cyclin A was determined by immunohistochemistry in paraffin sections of 126 soft tissue sarcomas. The median cyclin A score was 10.8% (range, 1-54%). Cyclin A expression correlated with the S-phase fraction, Ki-67 score, G2-M phase, and grade. It did not correlate with the size of the tumor. A high cyclin A score predicted a poor metastasis-free survival (P < 0.01) and a poor disease-specific overall survival (P = 0.01). We concluded that the expression of cyclin A is a powerful prognostic factor in soft tissue sarcoma. Moreover, the cyclin A score determines the fraction of tumor cells in the S phase and the G2 phase, which are the most sensitive cell cycle phases for current modalities of cancer treatment. PMID- 10383151 TI - The phosphatidylinositol 3'-kinase pathway is a dominant growth factor-activated cell survival pathway in LNCaP human prostate carcinoma cells. AB - Intracellular signaling pathways that mediate survival of prostate carcinoma (PCa) cells are poorly understood. We examined the potential role of the phosphatidylinositol 3' kinase (PI3K) pathway as a mediator of cell survival in LNCaP human PCa cells, which express a variety of properties characteristic of human prostate cancer. LNCaP cell cultures rapidly became apoptotic when treated with the specific PI3K inhibitors, wortmannin and LY294002. In contrast, apoptosis was not induced when the cells were treated with: (a) rapamycin, an inhibitor of the ribosomal S6 kinase pp70S6K, which acts downstream of PI3K; (b) PD98059, a specific inhibitor of the extracellular signal-regulated kinase/mitogen-activated protein kinase (Erk/MAPK) kinase (MEK); or (c) the antiandrogen, Casodex; or when the cells were cultured under androgen-depleted conditions. Apoptosis induced by PI3K inhibition was attenuated by: (a) dihydrotestosterone; or (b) the ErbB1 activating ligands [epidermal growth factor (EGF), transforming growth factor alpha, or heparin-binding EGF-like growth factor]. In response to ErbB1 activation by ligand, the p85 regulatory subunit of PI3K associated specifically with ErbB3 but not detectably with ErbB1. The anti apoptotic effect of ErbB1 activation was significantly reduced when cells were treated simultaneously with wortmannin and PD98059. These data indicate that survival signals can be evoked in LNCaP cells by several distinct pathways and can be triggered by nuclear and cell-surface receptors. Constitutive signaling through the PI3K pathway is required to prevent cell death in LNCaP, whereas activation of the Erk/MAPK and androgen response pathways is not obligatory for cell survival. These results also show that survival signals, as distinguished from mitogenic signals, can be evoked in PCa cells by ErbB1 ligands known to be synthesized within the human prostate. PMID- 10383152 TI - N-myc regulation of type I insulin-like growth factor receptor in a human neuroblastoma cell line. AB - Insulin-like growth factors I and II (IGF-I and IGF-II) stimulate proliferation and differentiation in many cell types, including cell lines derived from human neuroblastomas. Their effects are mediated via the IGF-I receptor (IGF-IR) that is essential for growth in these cells. Amplification of the N-myc oncogene is a marker for poor prognosis in neuroblastoma development, and it therefore seemed of interest to analyze the relationships that may exist between IGF-IR and N-myc. N-myc-deficient SK-N-SH neuroblastoma cells were used as an experimental model. After stable transfection with N-myc cDNA, Northern blotting revealed a marked increased in IGF-IR, IGF-II, IGF-binding protein (IGFBP)-2, and IGFBP-4 mRNA levels, whereas IGFBP-6 mRNA levels were clearly diminished. Western immunoblot analysis also demonstrated increased intact IGFBP-2 but decreased IGFBP-6 in the presence of N-myc oncogene. Parallel binding experiments using IGF-I missing the first 3 amino acids revealed a 47% increase in binding sites for IGF-I and an increase of at least 335% in DNA synthesis, as measured by labeled thymidine incorporation into DNA. s.c. injection of these cells into nude mice provoked xenograft development in 50-100% of cases (depending on the series of experiments). Control cells, in contrast, were not tumorigenic. In cells transfected with bp -420/+60 of the human IGF-IR promoter controlling expression of the luciferase reporter gene, promoter activity was stimulated by a factor of 3.8 +/- 0.6 (n = 6) in the presence of N-myc oncogene. This suggests transcriptional regulation of IGF-IR expression by N-myc. IGF-IR activity and N myc amplification are two events that to date have been identified as independently instrumental in the etiology of human neuroblastoma. Our results provide the first evidence of a direct link between them and demonstrate the effects of the oncogene on components of the IGF system in neuroblastoma cell growth in vitro and in vivo. PMID- 10383153 TI - Candidate genetic modifiers of individual susceptibility to renal cell carcinoma: a study of polymorphic human xenobiotic-metabolizing enzymes. AB - The steady increase in sporadic renal cell carcinoma (RCC) observed in industrialized countries supports the notion that certain carcinogens present in the environment (tobacco smoke, drugs, pollutants, and dietary constituents) may affect the occurrence of RCC. Many of the enzymes dealing with such environmental factors are polymorphic and may, therefore, confer variable susceptibility to RCC. This case-control study was designed to test for an association between genetic polymorphism of enzymes involved in xenobiotic metabolism and the risk of sporadic RCC. Genomic DNA was obtained from 173 patients with RCC and 211 controls of Caucasian origin. We used PCR-RFLP to investigate polymorphism for the most common alleles at two cytochrome-P450 mono-oxygenases (CYP1A1 and CYP2D6), one NAD[P]H:quinone oxidoreductase (NQO1), three glutathione S transferases (GSTM1, GSTT1, and GSTP1), and one N-acetyltransferase (NAT2) loci. The CYP1A1 (m) "variant" genotype, which contains at least one copy of the CYP1A1 variant alleles, was found to be associated with a 2.1-fold [95% confidence interval (CI), 1.1-3.9] increase in the risk of RCC. There was also a higher risk of RCC for subjects with the CYP1A1 (m) variant genotype combined with any of the following genotypes: GSTT1 (+) "active" [odds ratio (OR), 2.3; 95% CI, 1.2-4.5], GSTP1 (m) variant (OR, 2.4; 95% CI, 1.0-5.4), or NAT2 (-) "slow acetylator" (OR, 2.5; 95% CI, 1.1-5.5). A significant association was also found for the GSTM1 (-) "null" and GSTP1 (m) genotypes combined with either NAT2 (-) (OR, 2.6; 95% CI, 1.2-5.8) or CYP1A1 (m) (OR, 3.5; 95% CI, 1.1-11.2). The CYP2D6 (-) "poor metabolizer " and the NQO1 (-) "defective" genotypes were not clearly associated with a higher risk of RCC. Our data demonstrate for the first time a significant association between a group of pharmacogenetic polymorphisms and RCC risk. These positive findings suggest that interindividual variation in the metabolic pathways involved in the functionalization and detoxification of specific xenobiotics is an important susceptibility factor for RCC in Caucasians. PMID- 10383154 TI - Carcinoembryonic antigen (CEA)-specific T-cell activation in colon carcinoma induced by anti-CD3 x anti-CEA bispecific diabodies and B7 x anti-CEA bispecific fusion proteins. AB - Two bispecific recombinant molecules, an anti-CD3 x anti-carcinoembryogenic antigen (CEA) diabody and a B7 x anti-CEA fusion protein, were tested for their capacity to specifically activate T cells in the presence of CEA-expressing colon carcinoma cells. T-cell activation by the anti-CD3 x anti-CEA diabody required close contact to CEA-positive cells and resulted in diabody-mediated cytotoxicity against the target cells. Additionally, CD28-mediated costimulation in combination with anti-CD3 x anti-CEA diabodies induced activation of autologous T cells in CEA-positive primary colon carcinoma specimens, as determined by flow cytometry. The high specificity of the bispecific diabody approach could be further enhanced by the use of B7 x anti-CEA fusion proteins because the costimulatory CD28-signaling to the T cells strictly depended on the expression of CEA on the target cells. We demonstrate that displaying engagement sites for the T-cell antigens CD3 and CD28 on the surface of colon carcinoma cells is a suitable way to activate and retarget T cells in a highly tumor-specific manner. For clinical purposes, B7 x anti-tumor-associated antigen (TAA) fusion proteins, which are equally effective but more specific compared with anti-CD28 monoclonal anti-bodies, thus may improve the tumor specificity of anti-CD3 x anti-TAA bispecific antibodies. Furthermore, B7-negative tumors can be converted into B7 positive tumors by B7 x anti-TAA fusion proteins without the need for B7 gene transfer to the malignant cells. PMID- 10383155 TI - Tumor pretargeting with avidin improves the therapeutic index of biotinylated tumor necrosis factor alpha in mouse models. AB - The clinical use of tumor necrosis factor alpha (TNF) as an anticancer drug is limited to local or locoregional administration because of dose-limiting systemic toxicity. We investigated in animal models whether the therapeutic index of systemically administered human or murine TNF can be increased by tumor pretargeting strategies based on the biotin-avidin system. Pretargeting of s.c. mouse WEHI-164 fibrosarcoma and RMA lymphoma genetically engineered to express the Thy 1.1 antigen on the cell membrane was achieved by i.p. injection of a biotinylated anti-Thy 1.1 antibody and avidin. This pretreatment increased the antitumor activity of systemically administered biotin-TNF conjugates by at least 5-fold. In contrast, pretargeting did not increase the toxicity of biotin-TNF, as judged by animal survival and weight loss after treatment. Ex vivo analysis of tumor cells 24 h after treatment showed that biotin-TNF persisted for several hours on the surface of pretargeted tumors, but not when avidin was omitted. The potentiation of the antitumor effects was related primarily to indirect mechanisms, involving a host-mediated response. The results indicate that tumor pretargeting improves the antitumor activity of TNF. Tumor pretargeting with avidin, which is currently used to increase the uptake of radioactive-labeled biotin in patients, could represent a new strategy for improving the therapeutic index of TNF. PMID- 10383156 TI - Differential effects of a stem cell factor-immunoglobulin fusion protein on malignant and normal hematopoietic cells. AB - We genetically connected the extracellular domain of human stem cell factor to the Fc-portion of human IgG1. The chimeric recombinant stem cell factor IgG1 fusion protein (rSCF-IgG1) had an apparent approximately Mr 190,000 and consisted of three identical covalently linked subunits. It specifically bound to c-kit and the high affinity Fc gamma receptor, respectively. Liquid phase rSCF-IgG1 was, on a molar basis, about eight times more potent than native human rSCF in stimulating the proliferation of c-kit-positive leukemic cell lines and of nonmalignant CD34-positive hematopoietic progenitor cells. Although the effective dose conferring half maximum of [methyl-3H]thymidine uptake by liquid phase and solid phase-bound rSCF-IgG1 were comparable, the plateau level of [methyl 3H]thymidine uptake by malignant cells was decreased by the latter, whereas proliferation of nonmalignant progenitor cells was supported. Liquid phase rSCF IgG1 had a 2-fold increased potential to maintain primitive nonmalignant progenitor cells in stroma-free long-term culture compared with rSCF. Liquid phase rSCF-IgG1 caused enhanced and prolonged receptor phosphorylation and a more rapid down modulation of c-kit. Our data support the concept that solid phase attachment of rSCF-IgG1 is sufficient for alteration of biological function and that rSCF-IgG1 partially blocks SCF-stimulated malignant cell growth while supporting normal progenitor cells. PMID- 10383157 TI - KF25706, a novel oxime derivative of radicicol, exhibits in vivo antitumor activity via selective depletion of Hsp90 binding signaling molecules. AB - Radicicol, a macrocyclic antifungal antibiotic, has been shown to bind to the heat shock protein 90 (Hsp90) chaperone, interfering with its function. Hsp90 family chaperones have been shown to associate with several signaling molecules and play an essential role in signal transduction, which is important for tumor cell growth. Because radicicol lacks antitumor activity in vivo in experimental animal models, we examined the antitumor activity of a novel radicicol oxime derivative, radicicol 6-oxime (KF25706), on human tumor cell growth both in vitro and in vivo. KF25706 showed potent antiproliferative activities against various human tumor cell lines in vitro and inhibited v-src- and K-ras-activated signaling as well as radicicol. In addition, Hsp90 family chaperone-associated proteins, such as p185erbB2, Raf-1, cyclin-dependent kinase 4, and mutant p53, were depleted by KF25706 at a dose comparable to that required for antiproliferative activity. KF25706 was also shown to compete with geldanamycin for binding to Hsp90. KF29163, which is an inactive derivative of radicicol, was less potent both in p185erbB2 depletion and Hsp90 binding. More importantly, KF25706 showed significant growth-inhibitory activity against human breast carcinoma MX-1 cells transplanted into nude mice at a dose of 100 mg/kg twice daily for five consecutive i.v. injections. KF25706 was also shown to possess antitumor activity against human breast carcinoma MCF-7, colon carcinoma DLD-1, and vulval carcinoma A431 cell lines in vivo in an animal model. Finally, we confirmed the depletion of Hsp90-associated signaling molecules (Raf-1 and cyclin dependent kinase 4) with ex vivo Western blotting analysis using MX-1 xenografts. In agreement with in vivo antitumor activity, KF25706 depleted Hsp90-associated molecules in vivo, whereas KF29163 and radicicol did not show this activity in vivo. Taken together, these results suggest that antitumor activity of KF25706 may be mediated, at least in part, by binding to Hsp90 family proteins and destabilization of Hsp90-associated signaling molecules. PMID- 10383158 TI - Homocamptothecin, an E-ring modified camptothecin with enhanced lactone stability, retains topoisomerase I-targeted activity and antitumor properties. AB - Homocamptothecin (hCPT) is a semisynthetic analogue of camptothecin (CPT) with a seven-membered beta-hydroxylactone resulting from the insertion of a methylene spacer between the alcohol moiety and the carboxyl function of the naturally occurring six-membered alpha-hydroxylactone of CPT. This E-ring modification provides a less reactive lactone with enhanced stability and decreased protein binding in human plasma. Biological testing against CPT revealed that, instead of being detrimental, the modified lactone of hCPT has a positive impact on topoisomerase I (Topo I) poisoning properties. In vitro tests showed hCPT to fully conserve the ability to stabilize Topo I-DNA cleavage complexes and to stimulate a higher level of DNA cleavage than CPT. A similar trend toward improvement was also observed in antiproliferative assays with human tumor cell lines (including cells overexpressing P-glycoprotein). In two distinct in vivo models, using L1210 murine leukemia or human colon carcinoma HT29, hCPT was found to be more efficacious than CPT. The slow, but irreversible, hydrolysis of hCPT, instead of the fast equilibrium of CPT, may account for its good in vivo activity. Overall, these results provide evidence that a highly reactive lactone is not a requisite for the Topo I-mediated antitumor activity of CPT analogues, and that hCPT is an interesting pharmacological tool with improved solution behavior as well as a promising new template for the preparation of more efficacious Topo I poisons. PMID- 10383159 TI - Antitumor activity of P-4055 (elaidic acid-cytarabine) compared to cytarabine in metastatic and s.c. human tumor xenograft models. AB - The antineoplastic efficacy of P-4055, a 5'-elaidic acid (C18:1, unsaturated fatty acid) ester of cytarabine, a nucleoside antimetabolite frequently used in the treatment of hematological malignancies, was examined in several in vivo models for human cancer. In initial dose-finding studies in nude mice, the efficacy of P-4055 was highest when using schedules with repeated daily doses. In a Raji Burkitt's lymphoma leptomeningeal carcinomatosis model in nude rats, the control cytarabine- and saline-treated animals (five in each group) had a mean survival time of 13.2 days, whereas treatment with P-4055 resulted in three of five long-time survivors (>70 days). In a systemic Raji leukemia model in nude mice, 8 of 10 of the P-4055-treated animals survived (>80 days), compared with none of the cytarabine-treated animals (mean survival time, 34.2 days). In s.c. xenograft models, the effects of maximum tolerated doses of P-4055 and cytarabine, given in four weekly cycles of daily bolus i.v. injections for 5 subsequent days, against seven tumors (three melanomas, one lung adenocarcinoma, one breast cancer, and two osteogenic sarcomas) were investigated. P-4055 induced partial or complete tumor regression of the lung carcinoma, as well as of all three malignant melanomas. In two of the melanomas the activity was highly superior to that of cytarabine, and both P-4055 and cytarabine were, in general, more effective than several clinically established drugs previously tested in the same tumor models. In in vitro studies, inhibitors of nucleoside carrier dependent transport, nitrobenzylmercaptopurine riboside and dipyridamol, reduced strongly the cellular sensitivity to cytarabine, but not to P-4055, indicating that P-4055 uses an alternative/additional mechanism of internalization into the cell compared with cytarabine. The results explain, at least in part, the observed differences between the two compounds in in vivo efficacy, and together the data strongly support the evaluation of P-4055 in clinical studies. PMID- 10383160 TI - Primary chemically induced tumors induce profound immunosuppression concomitant with apoptosis and alterations in signal transduction in T cells and NK cells. AB - Whereas transplantable tumors can be readily cured with immunotherapeutic approaches, similar therapies in cancer patients have been less effective. This difference may be explained by an immunosuppression resulting from the presence of a slowly growing primary tumor in the patient, whereas the immune system in a mouse with a rapidly proliferating transplantable tumor would be less affected. As a more appropriate model to the immune dysfunction in patients, slowly progressing primary tumors were induced by the carcinogen methylcholanthrene (MC) in mice. Their ability to induce immunosuppression in T cells and natural killer (NK) cells was compared to that of rapidly growing transplanted MC-induced tumors. The results demonstrate that mice bearing primary MC tumors had significantly diminished T-cell and NK-cell functions, impaired capacity to produce Th1 cytokines, and markedly reduced levels of the signal-transducing zeta chain in T cells and NK cells, similar to that described in cancer patients. Moreover, a substantial number of CD8+ T cells in mice with large primary MC tumors were undergoing apoptosis, correlating with alterations in CD4/CD8 ratios. In contrast, T cells and NK cells from mice bearing rapidly growing transplanted tumors were only marginally affected. These findings could explain the apparent discrepancy between the consistent findings of a diminished immune response and alterations in signal transduction in cancer patients as compared to the less reproducible observations in murine transplantable tumors. In addition, they could explain the differences in the high efficacy of immunotherapy in mice with transplantable tumors and the low therapeutic results in cancer patients. PMID- 10383161 TI - p16/pRb pathway alterations are required for bypassing senescence in human prostate epithelial cells. AB - The cell cycle regulatory genes p16/CDKN2 and RB are frequently deleted in prostate cancers. In this study, we examined the role of alterations in p16 and pRb during growth, senescence, and immortalization in vitro of human prostate epithelial cells (HPECs). HPECs are established from normal prostate tissues and cultured on collagen-coated dishes. Our results show that p16 is reproducibly elevated at senescence in HPECs. HPECs are immortalized using human papilloma virus 16 E6 and/or E7 as molecular tools to inactivate p53 and/or pRb, respectively. Immortalization occurs infrequently in this system and only after a latent period during which additional genetic/epigenetic changes are thought to occur. Notably, all of the E6-immortalized HPEC lines but none of the E7 lines show inactivation of p16/CDKN2 (by deletion, methylation, or mutation) in association with immortalization. In contrast, E7 lines, in which pRb function is abrogated by E7 binding, retain the high levels of p16 observed at senescence. Thus, all lines show either a p16 or pRb inactivation. Analysis of six independent lines from metastatic prostate cancers reveals a similar loss of either p16 or pRb. Comparative genomic hybridization of HPECs shows that gains of chromosomes 5q, 8q, and 20 are nonrandomly associated with bypassing senescence (probability = 0.95). These results suggest that high levels of the cyclin dependent kinase inhibitor p16 mediate senescence G1 arrest in HPECs and that bypassing this block by a p16/pRb pathway alteration is required for immortalization in vitro and possibly tumorigenesis in vivo. Our results further indicate that inactivation of the p16/pRb pathway alone is not sufficient to immortalize HPECs and that additional genetic alterations are required for this process. PMID- 10383162 TI - Colonic hamartoma development by anomalous duplication in Cdx2 knockout mice. AB - To determine the biological role of caudal-like homeobox gene CDX2, we constructed knockout mice in which its mouse homologue Cdx2 was inactivated by homologous recombination, placing a bacterial lacZ gene under the control of the Cdx2 promoter. Although the homozygous mutants died in utero around implantation, the heterozygotes were viable and fertile and expressed lacZ in the caudal region in early embryos and in the gut tissues in adults. The heterozygotes developed cecal and colonic villi by anteriorization and formed hamartomatous polyps in the proximal colon. The hamartoma started to develop at 11.5 days of gestation as an outpocket of the gut epithelium, which ceased to express the remaining Cdx2 allele. The outpocket then expanded as a partially duplicated gut but was contained as a hamartoma after birth. In adult mice, these hamartomas grew very slowly and took a benign course. None of them progressed into invasive adenocarcinomas, even at 1.5 years of age. Whereas the cecal and colonic villi expressed lacZ, the hamartoma epithelium did not, nor did it express Cdx2 mRNA from the wild-type allele. However, genomic DNA analysis of the polyp epithelium did not show a loss of heterozygosity of the Cdx2 gene, suggesting a mechanism of biallelic Cdx2 inactivation other than loss of heterozygosity. These results indicate that the Cdx2 haploin-sufficiency caused cecal and colonic villi, whereas the biallelic inactivation of Cdx2 triggered anomalous duplications of the embryonic gut epithelium, which were contained as hamartomas after birth. PMID- 10383163 TI - Autocrine interleukin-1beta production in leukemia: evidence for the involvement of mutated RAS. AB - Interleukin (IL)-1beta is constitutively expressed in many leukemias and operates as an autocrine growth factor. To study the cellular basis for this aberrant production, we analyzed two cell lines, B1 (acute lymphoblastic leukemia) and W1 (juvenile chronic myelogenous leukemia), which express high levels of IL-1beta and have mutations in the K-RAS and N-RAS genes, respectively. Electromobility shift assays demonstrated transcription factor binding at multiple IL-1beta promoter elements [nuclear factor (NF)-IL6/CREB, NFB1, NFkappaB, and NF-IL6], consistent with the activation of an upstream signaling pathway. To determine whether activated Ras was involved, two structurally distinct classes of farnesyltransferase (FTase) inhibitors (the monoterpenes and a peptidomimetic) and an adenoviral vector expressing antisense targeted to K-RAS were used to specifically interfere with Ras function and/or expression. Treatment with the FTase inhibitors resulted in a concentration-dependent decrease in both NF IL6/CREB binding to the IL-1beta promoter and IL-1beta protein levels, without a significant change in total cellular protein levels. Furthermore, exposure of the B1 cells to antisense against K-RAS resulted in an approximately 50% reduction in both p21Ras and IL-1beta protein levels. Growth suppression was observed after FTase inhibitor or antisense exposure, an effect that was partially reversible by the addition of recombinant IL-1beta to the cultures. Our observations suggest that mutated RAS genes may mediate autocrine IL-1beta production in some leukemias by stimulating signal transduction pathways that activate the IL-1beta promoter. PMID- 10383164 TI - Carboxypeptidase A3 (CPA3): a novel gene highly induced by histone deacetylase inhibitors during differentiation of prostate epithelial cancer cells. AB - Butyrate and its structural analogues have recently entered clinical trials as a potential drug for differentiation therapy of advanced prostate cancer. To better understand the molecular mechanism(s) involved in prostate cancer differentiation, we used mRNA differential display to identify the gene(s) induced by butyrate. We found that the androgen-independent prostate cancer cell line PC-3 undergoes terminal differentiation and apoptosis after treatment with sodium butyrate (NaBu). A novel cDNA designated carboxypeptidase A3 (CPA3), which was up-regulated in NaBu-treated PC-3 cells, was identified and characterized. This gene expresses a 2795-bp mRNA encoding a protein with an open reading frame of 421 amino acids. CPA3 has 37-63% amino acid identity with zinc CPs from different mammalian species. It also shares 27-43% amino acid similarity with zinc CPs from several nonmammalian species, including Escherichia coli, yeast, Caenorhabditis elegans, and Drosophila. The structural similarity between CPA3 and its closest homologues indicates that the putative CPA3 protein contains a 16 residue signal peptide sequence, a 95-residue NH2-terminal activation segment, and a 310-residue CP enzyme domain. The consistent induction of CPA3 by NaBu in several prostate cancer cell lines led us to investigate the signaling pathway involved in the induction of CPA3 mRNA. Trichostatin A, a potent and specific inhibitor of histone deacetylase, also induced CPA3 mRNA expression, suggesting that CPA3 gene induction is mediated by histone hyperacetylation. We demonstrated that CPA3 induction was a downstream effect of the treatment with butyrate or trichostatin A, but that the induction of p21(WAF1/CIP1) occurred immediately after these treatments. We also demonstrated that the induction of CPA3 mRNA by NaBu was inhibited by p21(WAF1/CIP1) antisense mRNA expression, indicating that p21 transactivation is required for the induction of CPA3 by NaBu. Our data demonstrate that the histone hyperacetylation signaling pathway is activated during NaBu-mediated differentiation of PC-3 cells, and the new gene, CPA3, is involved in this pathway. PMID- 10383165 TI - Heparin/heparan sulfate interacting protein gene expression is up-regulated in human colorectal carcinoma and correlated with differentiation status and metastasis. AB - We applied a subtractive hybridization strategy to obtain genes that are differentially expressed in colorectal carcinoma. Heparin/heparan sulfate interacting protein (HIP) was shown to be up-regulated in colorectal carcinoma. A study of 53 patients with documented colorectal carcinoma showed that 70% of the tumors had HIP tumor-to-normal ratios (expression in tumor tissue compared to expression in normal mucosa) of >2. In six patients with concomitant polyps, HIP expression in the polyps was similar to the carcinoma, showing that up-regulation of HIP may be an early event in tumorigenesis. A significant inverse correlation between HIP levels and the presence of distant metastasis (Duke's stage D) was noted. Similarly, HIP expression was also related to differentiation status in human colorectal carcinoma cell lines. HIP expression was lower in the poorly differentiated COLO 205 cell line compared to the well-differentiated HT-29 cell line. The correlation was further strengthened by studies in COLO 205 cells that were induced to differentiate with herbimycin A treatment. HIP expression was significantly higher when the cells were induced to differentiate. Withdrawal of herbimycin A resulted in a reversal of morphological changes associated with differentiation and an associated decrease in HIP expression. These studies indicate that HIP is an important molecule for cell-cell and cell-extracellular matrix adhesion. The up-regulation of HIP may be an early event in tumorigenesis, and its increased expression may facilitate growth and local invasion. A lower expression of HIP in tumors results in decreased cell adhesion, favoring metastasis. HIP is a candidate marker of abnormal cell growth in the colon and a prognostic marker for colorectal carcinoma. PMID- 10383166 TI - Frameshift mutations at mononucleotide repeats in caspase-5 and other target genes in endometrial and gastrointestinal cancer of the microsatellite mutator phenotype. AB - The majority of tumors from hereditary nonpolyposis colorectal cancer families and a subset of unselected gastrointestinal and endometrial tumors exhibit a microsatellite mutator phenotype (MMP) that leads to the accumulation of hundreds of thousands of clonal mutations in simple repeat sequences. The mutated genes with positive or negative roles in cell growth or survival in aneuploid gastrointestinal cancer (e.g., APC, K-ras, and p53) are less frequently mutated in near-diploid MMP gastrointestinal tumors. These tumors accumulate mutations in other genes, such as DNA mismatch repair hMSH3 and hMSH6, transforming growth factor-beta type II receptor, and BAX. All these genes carry, within their coding sequences, mononucleotide repeats that are preferred targets for the MMP. Endometrial carcinoma is the most common type of extracolonic neoplasia in the hereditary nonpolyposis colorectal cancer syndrome, but the spectrum of its target cancer genes is not well characterized. Here, we report that endometrial cancer of the MMP also accumulates mutations in genes that are typically mutated in gastrointestinal cancer of the mutator pathway, including BAX (55%), hMSH3 (28%), and hMSH6 (17%). We also report the detection of frameshift mutations in caspase-5, a member of the caspase family of proteases that has an (A)10 repeat within its coding region, in MMP tumors of the endometrium, colon, and stomach (28, 62, and 44%, respectively). We therefore suggest caspase-5 as a new target gene in the microsatellite mutator pathway for cancer. PMID- 10383167 TI - Proteomics and immunohistochemistry define some of the steps involved in the squamous differentiation of the bladder transitional epithelium: a novel strategy for identifying metaplastic lesions. AB - Here, we present a novel strategy for dissecting some of the steps involved in the squamous differentiation of the bladder urothelium leading to squamous cell carcinomas (SCCs). First, we used proteomic technologies and databases (http://biobase.dk/cgi-bin/celis) to reveal proteins that were expressed specifically by fresh normal urothelium and three SCCs showing no urothelial components. Thereafter, antibodies against some of the differentially expressed proteins as well as a few known keratinocyte markers were used to stain serial cryostat sections (immunowalking) of biopsies obtained from bladder cystectomies of two of the SCC-bearing patients (884-1 and 864-1). Because bladder cancer is a field disease, we surmised that the urothelium of these patients may exhibit a spectrum of abnormalities ranging from early metaplastic stages to invasive disease. Immunohistochemical analysis revealed three types of non-keratinizing metaplastic lesions (types 1-3) that did not express keratins 7, 8, 18, and 20 (expressed by normal urothelium) and could be distinguished based on their staining with keratin 19 antibodies. Type 1 lesions showed staining of all cell layers in the epithelium (with differences in the staining intensity of the basal compartment), whereas type 2 lesions exhibited mainly basal cell staining. Type 3 lesions did not stain with keratin 19 antibodies. In cystectomy 884-1, type 3 lesions exhibited the same immunophenotype as the SCC and may be regarded as precursors to the tumor. Basal cells in these lesions did not express keratin 13, suggesting that the tumor, which was also keratin 13 negative, may have arisen from the expansion of these cells. Similar results were observed with cystectomy 864-1, which showed carcinoma in situ of the SCC type. SCC 864-1 exhibited both keratin 19-negative and -positive cells, implying that the tumor arose from the expansion of the basal cell compartment of type 2 and 3 lesions. Besides providing with a novel strategy for revealing metaplastic lesions, our studies have shown that it is feasible to apply powerful proteomic technologies to the analysis of complex biological samples under conditions that are as close as possible to the in vivo situation. PMID- 10383168 TI - The viral protein Apoptin induces apoptosis in UV-C-irradiated cells from individuals with various hereditary cancer-prone syndromes. AB - Apoptin, a protein derived from chicken anemia virus, has previously been shown to induce apoptosis in a p53-independent and Bcl-2-stimulated manner in transformed and tumorigenic human cells but not in normal diploid human cells, suggesting that it is a potential agent for tumor therapy. Here we report that Apoptin can induce apoptosis in UV-C-irradiated diploid skin fibroblasts from individuals with various hereditary cancer-prone syndromes that are characterized by a germ-line mutation in a tumor suppressor gene. The same effect is found when these cells are irradiated with X-rays. In contrast, diploid skin fibroblasts from healthy donors or from individuals with DNA repair disorders are not responsive to Apoptin-induced apoptosis upon UV-C or X-ray irradiation. After transfection of untreated cells, Apoptin is found predominantly in the cytoplasm, whereas in UV-C-exposed Apoptin-responsive cancer-prone cells, it migrates to the nucleus, where it causes rapid apoptosis. Apoptin remains localized in the cytoplasm after UV-C treatment of diploid cells from healthy individuals. The induction of apoptosis by Apoptin in cancer-prone cells with a germ-line mutation in a tumor suppressor gene is UV dose-dependent and transient, just like many other UV-induced processes. These results suggest that Apoptin may be used as a diagnostic tool for detection of individuals with an increased risk for hereditary cancer and premalignant lesions. PMID- 10383169 TI - A brief history of psychiatry: millennia past and present. AB - As a new millennium dawns, it is worth looking at where we have been and where we are going. In an age in which the only constant is change, the only new thing, in fact, is technology. In psychiatry there are promises of advances in diagnosis with search of the human genome, in the development of newer pharmacological agents through molecular biology, and in better understanding of psychosocial treatment using sophisticated contemporary research techniques. These represent extensions of theories about mind, disease, and treatment reaching back into antiquity. Many excellent books and a multiplicity of articles have been published in the 1990s about the history of all of these areas. This series of four articles will provide a selective summary of that literature and an up-to date guide to the history of psychiatry. PMID- 10383170 TI - Postpartum period: a risk factor for neuroleptic malignant syndrome. AB - Neuroleptic malignant syndrome (NMS) occurred in four postpartum patients treated with various psychotropic medications including depot-neuroleptics over a 5-year period. All four patients had the onset of NMS in the first few weeks of the postpartum period. Whether or not there is an increased risk for the development of NMS in the puerperium period is discussed. PMID- 10383171 TI - Hypofrontal symptoms from olanzapine: a case report. AB - Olanzapine acutely induced disabling hypofrontal symptoms in a 31-year-old male. This occurred after 13 years of exposure to typical neuroleptics without such symptoms. Presumably, hypofrontal symptoms should limit the dose of atypical neuroleptics in some patients. Milder expressions of hypofrontal symptoms should be more common. PMID- 10383172 TI - Neuropsychiatric aspects of carbon monoxide poisoning: a review and single case report suggesting a role for amphetamines. AB - Sublethal exposure to carbon monoxide (CO) can result in severe neurologic and psychiatric complications. Once in the body, CO can wreak havoc on virtually every organ system, with the brain being the most vulnerable to the damaging effects. Neuropathological injury is frequently widespread, and while white matter injury is most common, both gray and white matter injury occurs. Consequently, no neurologic or psychiatric syndrome is pathognomonic for CO poisoning. There are currently no effective treatments for the delayed neuropsychiatric sequelae of CO poisoning, and medical management focuses on correcting immediate symptoms through the use of oxygen, hyperbaric oxygen therapy, and supportive measures. Preliminary data suggest, however, that dopaminergic agents may be useful for the treatment of some of the delayed sequelae of CO neurotoxicity. To our knowledge, ours is the first case report in which dextroamphetamine (DAMP), a potent dopaminergic agent, has been used for treating the neuropsychiatric symptoms of CO poisoning. Our data demonstrate that it is effective in shortening cognitive and motor recovery time, that psychostimulant actions occur slightly sooner than locomotor effects, and that theraputic benefit is most dramatic within the first ten days of use. Therefore, DAMP appears to be a pharmacological agent that can be combined with supportive interventions to reverse, attenuate, or symptomatically improve the delayed sequelae that occur in these patients. PMID- 10383173 TI - Pet-facilitated therapy for posttraumatic stress disorder. AB - It is suggested that pet-facilitated therapy might be a useful adjuvant on treatment of posttraumatic stress disorder. Some motivation and rationale for this idea is given, and a discussion of method of testing it. PMID- 10383174 TI - Then and now: ANZBA. Past, present, future: some personal recollections and observations on the Association and the development of burn care in Australia. PMID- 10383175 TI - Developing valid benchmarks for laboratories and regulatory agencies. PMID- 10383177 TI - Selective guide to current reference sources on topics discussed in this issue. PMID- 10383176 TI - Proceedings of the 25th Anniversary of the National Association for the Relief of Paget's Disease, International Symposium. Oxford, United Kingdom, 15-17 July 1998. PMID- 10383178 TI - Selective guide to current reference sources on topics discussed in this issue. PMID- 10383179 TI - Selective guide to current reference sources on topics discussed in this issue. PMID- 10383180 TI - Selective guide to current reference sources on topics discussed in this issue. PMID- 10383181 TI - Selective guide to current reference sources on topics discussed in this issue. PMID- 10383182 TI - Selective guide to current reference sources on topics discussed in this issue. Intensive outpatient treatment for the addictions. PMID- 10383183 TI - Selective guide to current reference sources on topics discussed in this issue. PMID- 10383184 TI - Selective guide to current reference sources on topics discussed in this issue. Integration of pharmacological and nonpharmacological treatments in drug/alcohol addictions. PMID- 10383185 TI - Selective guide to current reference sources on topics discussed in this issue. PMID- 10383186 TI - Selective guide to current reference sources on topics discussed in this issue. PMID- 10383187 TI - Selective guide to current reference sources on topics discussed in this issue. PMID- 10383188 TI - Selective guide to current reference sources on topics discussed in this issue. PMID- 10383189 TI - Selective guide to current reference sources on topics discussed in this issue. PMID- 10383190 TI - Abciximab binding to glycoprotein IIb-IIa and protein tyrosine phosphorylation in human platelets. PMID- 10383191 TI - Description of a new mutation in the L-ferrin iron-responsive element associated with hereditary hyperferritinemia-cataract syndrome in a Spanish family. PMID- 10383192 TI - A single mutation inside the NPA motif of aquaporin-1 found in a Colton-null phenotype. PMID- 10383193 TI - Periodic hematological disorders. PMID- 10383194 TI - No association between factor V Leiden and C282Y mutation in the hereditary hemochromatosis gene. PMID- 10383195 TI - Nuclear localization of the Fanconi anemia protein FANCC is required for functional activity. PMID- 10383196 TI - Evidence for continuous basel generation of GC-MAF: absence in infantile osteopetrosis and restoration after bone marrow transplant. PMID- 10383197 TI - The structure in solution of the b domain of protein disulfide isomerase. AB - Protein disulfide isomerase (PDI) is a multifunctional protein of the endoplasmic reticulum, which catalyzes the formation, breakage and rearrangement of disulfide bonds during protein folding. It consists of four domains designated a, b, b and a. Both a and a domains contains an active site with the sequence motif -Cys-Gly His-Cys-involved directly in thiol-disulfide exchange reactions. As expected these domains have structures very similar to the ubiquitous redox protein thioredoxin. A low-resolution NMR structure of the b domain revealed that this domain adopts a fold similar to the PDI a domain and thioredoxin [Kemmink, J., Darby, N.J., Dijkstra, K., Nilges, M. and Creighton, T.E. (1997) Curr. Biol. 7, 239-245]. A refined ensemble of solution structures based on the input of 1865 structural restraints shows that the structure of PDI b is well defined throughout the complete protein except for about 10 residues at the C-terminus of the sequence. 15N relaxation data show that these residues are disordered and not part of this structural domain. Therefore the domain boundaries of PDI can now be fixed with reasonable precision. Structural comparison of the PDI b domain with thioredoxin and PDI a reveals several features important for thiol-disulfide exchange activity. PMID- 10383198 TI - A robust and cost-effective method for the production of Val, Leu, Ile (delta 1) methyl-protonated 15N-, 13C-, 2H-labeled proteins. AB - A selective protonation strategy is described that uses [3-2H] 13C alpha ketoisovalerate to introduce (1H-delta methyl)-leucine and (1H-gamma methyl) valine into 15N-, 13C-, 2H-labeled proteins. A minimum level of 90% incorporation of label into both leucine and valine methyl groups is obtained by inclusion of approximately 100 mg/L alpha-ketoisovalerate in the bacterial growth medium. Addition of [3,3-2H2] alpha-ketobutyrate to the expression media (D2O solvent) results in the production of proteins with (1H-delta1 methyl)-isoleucine (> 90% incorporation). 1H-13C HSQC correlation spectroscopy establishes that CH2D and CHD2 isotopomers are not produced with this method. This approach offers enhanced labeling of Leu methyl groups over previous methods that utilize Val as the labeling agent and is more cost effective. PMID- 10383200 TI - Neuromuscular disorders: gene location. PMID- 10383199 TI - Efficient determination of angles subtended by C(alpha)-H(alpha) and N-H(N) vectors in proteins via dipole-dipole cross-correlation. AB - The angle Theta(C(alpha)H(alpha)),NH(N) subtended by the internuclear vectors 13C(alpha)-H(alpha) and 15N-H(N) in doubly-labeled proteins can be determined by observing the effect of cross-correlation between the dipolar interactions on zero- and double-quantum coherences involving 13C(alpha) and 15N. Two complementary 2D experiments with the appearance of 15N-HN correlation spectra yield signal intensities that depend on the rate of interconversion through cross correlated relaxation of in-phase and doubly antiphase zero- and double-quantum coherences. The ratio of the signal intensities in the two experiments bears a simple relationship to the cross-correlation rate, and hence to the angle Theta(C(alpha)H(alpha),NH(N)). Assuming planarity of the peptide bond, the dihedral angle psi (between C(alpha) and C) can be determined from the knowledge of Theta(C(alpha)H(alpha),NH(N)). The experiments are very time-effective and provide good sensitivity and excellent spectral resolution. PMID- 10383201 TI - Mitochondrial encephalomyopathies: gene mutation. PMID- 10383202 TI - [Annual Meeting and Continuing Education Congress of OGGG and Scientific Meeting of the Austrian Society of Prenatal and Perinatal Medicine]. PMID- 10383203 TI - Commentary:plausible candidates for treatment of glue ear-is one issue really three? PMID- 10383204 TI - Early discharge after surgery for breast cancer. Hotel wards are good option for patients. PMID- 10383205 TI - Early discharge after surgery for breast cancer. More evidence in favour of early discharge. PMID- 10383206 TI - Guidelines in general practice. Problem is that guidelines are useful. PMID- 10383207 TI - Guidelines in general practice. Post of primary care knowledge officer could be set up. PMID- 10383208 TI - More on albumin. Multicentre randomised controlled trail is needed before changing resuscitation formulas for major burns. PMID- 10383209 TI - Organisational and cultural aspects are also important. PMID- 10383210 TI - Intranasal corticosteroids in allergic rhinitis. Evidence of efficacy is useful but not only factor to be considered. PMID- 10383211 TI - Memories of why allocation by random sampling numbers was used. PMID- 10383212 TI - Public health psychiatry and crime prevention. Psychiatry cannot protect public from people with personality disorder. PMID- 10383213 TI - Cushing's syndrome induced by betamethasone nose drops. Children taking intranasal corticosteroids should be monitored for growth retardation. PMID- 10383214 TI - Cushing's syndrome induced by betamethasone nose drops. Learning difficulties may complicate the issue. PMID- 10383215 TI - Does the fly matter in trout fishing? Trout in trout fisheries in New England have different interests. PMID- 10383216 TI - Underwater and Hyperbaric Medicine, abstracts from the literature. PMID- 10383217 TI - Asking difficult questions. PMID- 10383218 TI - Capillary electrophoresis of some tetracycline antibiotics coupled with reductive fast cyclic voltammetric detection. AB - The separation and quantitative performance parameters for tetracycline, chlortetracycline and oxytetracycline antibiotics were investigated by capillary zone electrophoresis coupled with fast cyclic voltammetric detection. Optimization of pH and complexation with a boric acid-sodium tetraborate buffer provided good resolution of all compounds. Detection by electrochemical reduction using fast on-line cyclic voltammetric detection with a Hg-film-microm electrode gave detection limits (2 x peak-to-peak baseline noise) of 7 x 10(-7) mol/l for tetracycline and chlortetracycline, and 1.5 x 10(-6) mol/l for oxytetracycline. The influence of electrode material, potential range and scan rate was examined and discussed. Optimal electrochemical detection was obtained at a Hg-film electrode with a waveform that consisted of an initial constant potential of -0.6 V for 200 ms followed by a cyclic voltammetry (CV) scan at 300 V/s from - 0.6 V to a vertex potential of 1.7 V. The analytical signal was obtained by plotting the integrated values of the CV current from each applied waveform as a function of time. The calibration plot (peak areas) for each separated peak was found to be linear over three-orders of magnitude. PMID- 10383219 TI - Determination of alpha-tocopherol and alpha-tocopherol acetate in diets of experimental animals. Study of stability in the diets. AB - A simple method is described which permits, avoiding saponification, alpha tocopherol and alpha-tocopheryl acetate measurement in semi-synthetic diets for experimental animals by HPLC, with both UV and fluorescence detection. Phenyldodecane was chosen as internal standard with remarkable performances, and EDTA and BHT were added to prevent oxidation in aqueous and non-aqueous phases respectively. The mobile phase was methanol-water (94:6 v/v) at a flow-rate of 2 ml/min. Samples were homogenized and extracted twice with n-hexane by probe sonication. Extracts were evaporated to dryness and redissolved with chloroform methanol (1:1, v/v). Validation parameters were studied between 25 ng and 6 micrograms for alpha-tocopherol and between 3 and 24.2 micrograms for alpha tocopheryl acetate, which corresponds to the range of values in the existing diets. Results had correlation coefficients > 0.99; recoveries > 85%; R.S.D. < 6%, so the method is adequate to control vitamin E intake in animals as well as vitamin E stability in food during storage. PMID- 10383220 TI - Ethnic factors: implications for drug therapy and global drug development. AGAH annual meeting, Heidelberg, 7-9 February 1999. Abstracts. PMID- 10383221 TI - Understanding Bacillus anthracis pathogenesis. PMID- 10383222 TI - Evolution of Helicobacter pylori: the role of recombination. PMID- 10383223 TI - Molecular mechanism of nerve infection in leprosy. PMID- 10383224 TI - When microbes and cells meet. PMID- 10383225 TI - The introduction of tincture of iodine sterilization. PMID- 10383226 TI - Isolation of a methanogenic bacterium, Methanosarcina sp. strain FR, for its ability to degrade high concentration of perchloroethylene. AB - Tetrachloroethylene (PCE) is a toxic compound essentially used as a degreasing and dry-cleaning solvent. A methanogenic and sulfate-reducing consortium that dechlorinates and mineralizes high concentrations of PCE was derived from anaerobically digested sludge obtained from a waste water treatment plant (Bourg en-Bresse, France). A methanogenic bacterium, strain FR, was isolated from this acclimated consortium. On the basis of morphological and physiological characteristics, strain FR was classified in the genus of Methanosarcina. Phylogeny analysis with the 16S rRNA gene sequence revealed that strain FR is highly related to Methanosarcina mazei and Methanosarcina frisia (99.6 and 99.5% identity, respectively). High concentrations (50-87 microM) of PCE were completely dechlorinated by strain FR cultures at the rate of 76 nM-mg protein( 1).day(-1). PCE dechlorination produced a nonidentified compound. The tracer experiments with [13C]PCE revealed that the product was nonchlorinated. Dechlorination of PCE to trichloroethylene was still active in the presence of boiled cell extract of the strain FR. However, no further dechlorination was observed. This result suggests that a cofactor rather than an enzymatic system is responsible for the first dechlorination of PCE. Dechlorination-active fractions purified from cell extracts on a XAD-4 column revealed the presence of F(420), F(430), and cobamides cofactors. This is the first report of the isolation of a methanogenic bacterium with the ability to dechlorinate high concentrations of PCE to a nonchlorinated product. PMID- 10383227 TI - Aging of Cryptosporidium parvum oocysts in river water and their susceptibility to disinfection by chlorine and monochloramine. AB - Cryptosporidium parvum oocysts were aged in waters from both the St. Lawrence River and the Ottawa River. In situ survival experiments were carried out by incubating the oocysts in either dialysis cassettes or microtubes floated into an overflow tank. A significant portion of the oocysts survived in the test waters for several weeks. Oocyst survival in the St. Lawrence River was better in membrane-filtered (0.2 microm-pore diameter) water than in unfiltered water, suggesting that biological antagonism may play a role in the environmental fate of the parasite. Oocysts aged in river waters under in situ conditions and control oocysts kept refrigerated in synthetic water (100 ppm as CaCO3); pH 7.0) were subjected to the same disinfection protocol. Aged oocysts were at least as resistant as, if not more resistant than, the control oocysts to disinfection. This indicates that the oocysts surviving in the water environment may be just as difficult to inactivate by potable water disinfection as freshly shed oocysts. Therefore, water treatment should not be based on the assumption that environmental oocysts may be more easily inactivated than freshly shed oocysts. First-order kinetics die-off rates varied from one river to another (from 0.013 to 0.039 log(10).day(-1)) and from one experiment to another with water from the same river collected at different times. Calculation of the die-off rates based on either in vitro excystation or in vitro excystation in combination with total counts (overall die-off rates) showed that the assessment of oocyst viability by microscopic methods must account for the total oocyst loss observed during long term inactivation assays of river waters. PMID- 10383228 TI - Delicious vaccines. Axis Genetics, plc. PMID- 10383229 TI - Cottle--The individual remarks made during the opening ceremonies of the Cottle International Rhinology Centennial in Philadelphia, June 4, 1997. PMID- 10383230 TI - Proceedings of the 7th International Congress on the Neuronal Ceroid Lipofuscinoses: NCL-98. Dallas, Texas, USA. June 13-16, 1998. PMID- 10383231 TI - Alternative and complementary medicine. PMID- 10383233 TI - Microbial Genomes III: Sequencing, Functional Characterization and Comparative Genomics. Chantilly, Virginia, USA. January 29-February 1, 1999. Abstracts. PMID- 10383232 TI - [In vivo monitoring of chemotherapy multidrug resistance (MDR)]. PMID- 10383234 TI - NIH consensus panel recommends expanding access to and improving methadone treatment programs for heroin addiction. PMID- 10383235 TI - Cell regulation. Web alert. PMID- 10383236 TI - Royal College of Obstetricians and Gynaecologists Evidence-based Clinical Guidelines. Guideline Summary No. 3: the management of infertility in secondary care. PMID- 10383237 TI - Holmium:YAG lasertripsy for ureteric calculi: an experience of 300 procedures. PMID- 10383238 TI - Haematuria and clot retention after transurethral resection of the prostate: a pilot study. PMID- 10383239 TI - Haematuria and clot retention after transurethral resection of the prostate: a pilot study. PMID- 10383240 TI - The hormonal assessment of the infertile male. PMID- 10383241 TI - Festschrift honoring Robert U. Massey, M.D. Professor, Dean, and Editor-Emeritus. PMID- 10383242 TI - The transcription factor GATA6 is essential for early extraembryonic development. AB - The gene coding for the murine transcription factor GATA6 was inactivated by insertion of a beta-galactosidase marker gene. The analysis of heterozygote GATA6/lacZ mice shows two inductions of GATA6 expression early in development. It is first expressed at the blastocyst stage in part of the inner mass and in the trophectoderm. The second wave of expression is in parietal endoderm (Reichert's membrane) and the mesoderm and endoderm that form the heart and gut. Inactivation leads to a lethality shortly after implantation (5.5 days postcoitum). Chimeric experiments show this to be caused by an indirect effect on the epiblast due to a defect in an extraembryonic tissue. PMID- 10383243 TI - Preparing for the new millennium. PMID- 10383244 TI - The phytoestrogen content of rodent diets. PMID- 10383245 TI - The TEF approach for hexachlorobenzene. PMID- 10383246 TI - Help stopping smoking. PMID- 10383248 TI - Preferable products mean a healthier Earth. PMID- 10383247 TI - Early action on climate change. PMID- 10383249 TI - Synchronous unilateral parotid neoplasms of different histological types. AB - The occurrence of multiple tumours in the salivary glands is an unusual phenomenon and the simultaneous development of tumours different types is extremely rare. Two cases are presented with synchronous tumours of the parotid gland of different histological types. The first was a Warthin tumour in combination with a metastatic lung carcinoma and the second was a pleomorphic adenoma in combination with non-Hodgkin's malignant lymphoma. PMID- 10383250 TI - Carcinosarcoma of the submandibular salivary gland. AB - We report a rare case of submandibular salivary gland carcinosarcoma ('true' malignant mixed tumour) which occurred in a 77-year-old man. Microscopic examination showed a neoplasm comprised of sarcomatous elements (chondrosarcoma, rhabdomyosarcoma and osteosarcoma) with tabular salivary ductal adenocarcinoma. A short review of the literature is also presented and the poor prognosis of these tumours, in spite of complete surgical removal and additional radiation therapy and chemotherapy, is discussed. PMID- 10383251 TI - Biliary cystadenoma: an uncommon cause of cholestatic jaundice. PMID- 10383253 TI - [Findings on the status of bacteria in nature and its effects]. PMID- 10383252 TI - Recurrent adrenocortical carcinoma in a child. AB - An 8-month-old girl presented with clitoromegaly, cushingoid features and a large abdominal tumour. Ultrasonography (US) and computed tomography (CT) of the abdomen revealed a tumour of the left suprarenal gland, 12x11x7 cm in size. Serum levels of cortisol, testosterone and DHEA-S, and urinary extretion of 17 ketosteroids and 17-hydroxycorticoids were increased. Complete removal of the tumour was accomplished through a transabdominal approach. The diagnosis of adrenocortical carcinoma was confirmed histologically. Three months after the first operation, a recurrent tumour of the left renal hilus, 23x15 mm in size, was identified by US and verified by aspiration biopsy. The tumour was removed by the transabdominal route. In this report, we discuss the diagnosis and the treatment of this rare disease. PMID- 10383254 TI - Acarbose and acetaminophen- a dangerous combination? PMID- 10383255 TI - Proceedings of the single theme consensus conference on Hepatitis B and C: Carrier to Cancer, Asian Perspectives. New Delhi, India, December 5-6, 1998. PMID- 10383256 TI - Economic reform in private veterinary practice. PMID- 10383257 TI - Rapid immunochromatographic assay for diagnosis of tuberculosis: antibodies detected may not be specific. PMID- 10383258 TI - cagA and vacA status of Spanish Helicobacter pylori clinical isolates. PMID- 10383259 TI - Interpretation of pneumococcal susceptibility tests with cefaclor. PMID- 10383260 TI - Classification of hepatitis C virus type 2 isolates by phylogenetic analysis of core and NS5 regions. PMID- 10383262 TI - Ideal carrier particles for agglutination tests. PMID- 10383261 TI - Treatment strategies for group A streptococcal pharyngitis. PMID- 10383263 TI - Phenylpropionic acid metabolism: a marker for enteropathogenic Escherichia coli clonal group 2 strains. PMID- 10383264 TI - Ability of the modified Vitek card to detect coagulase-negative staphylococcal with mecA and Oxacillin-resistant phenotypes. PMID- 10383265 TI - Report of the BIPP Medical Group/Scottish Region study day on 'the changing face of communications', held at Glasglow Royal Infirmary on Sunday 5th April 1998. PMID- 10383266 TI - Off-pump internal thoracic artery-left anterior descending coronary artery grafting via small anterior thoracotomy: when and compared to what? PMID- 10383267 TI - The use of the calcium antagonist nicardipine in arterial coronary bypass surgery. PMID- 10383268 TI - Regional cerebral blood flow and regional glucose use during rewarding after hypothermic cardiopulmonary bypass. PMID- 10383269 TI - Coronary artery bypass grafting with the descending branch of the lateral femoral circumflex artery used as an arterial conduit: is arteriographic evaluation necessary before its use? PMID- 10383270 TI - A successful treatment of serous leakage from a polytetrafluoroethylene Blalock Taussig shunt with intravenous fibrinogen administration. PMID- 10383271 TI - Aortic function in patients during intra-aortic balloon pumping determined by the pressure-diameter relation. PMID- 10383272 TI - Myocardial pseudovascular tubes are present in the delayed rejection of pig-to baboon orthotopic cardiac xenografts. PMID- 10383273 TI - Etanercept in rheumatoid arthritis. PMID- 10383274 TI - Etanercept in rheumatoid arthritis. PMID- 10383275 TI - Etanercept in rheumatoid arthritis. PMID- 10383276 TI - Respiratory viruses and acute otitis media. PMID- 10383277 TI - Respiratory viruses and acute otitis media. PMID- 10383278 TI - Radiotherapy for small-cell lung cancer. PMID- 10383279 TI - Radiotherapy for small-cell lung cancer. PMID- 10383280 TI - Radiotherapy for small-cell lung cancer. PMID- 10383281 TI - Pure red-cell aplasia. PMID- 10383282 TI - Pure red-cell aplasia. PMID- 10383283 TI - Gastropathy due to celecoxib, a cyclooxygenase-2 inhibitor. PMID- 10383284 TI - A Pepsi challenge. PMID- 10383285 TI - Comparing hospitals. PMID- 10383286 TI - Comparing hospitals. PMID- 10383287 TI - A word to the wise. PMID- 10383288 TI - An eye-opener on migraines. PMID- 10383289 TI - Infant mortality and low birthweight in North Carolina: the last 10 years. PMID- 10383290 TI - Could we keep the rabies epidemic away from the Outer Banks? Oral rabies vaccination of raccoons. PMID- 10383291 TI - Childhood constipation and encopresis: diagnosis and management. PMID- 10383292 TI - Internet courses for off-campus students. PMID- 10383293 TI - It came from the deep. PMID- 10383294 TI - Getting wired. New observations may show how neurons form connections. PMID- 10383295 TI - When good health is good business. PMID- 10383296 TI - Seeing the breath of life. PMID- 10383297 TI - New date for the dawn of dream time. PMID- 10383298 TI - AIDS center shuttered. PMID- 10383299 TI - Shutdown of research at Duke sends a message. PMID- 10383300 TI - Australia. Budget back report on boosting biotech. PMID- 10383301 TI - Subjecting belief to the scientific method. PMID- 10383303 TI - Jaw origins. PMID- 10383302 TI - Nuclear transport protein does double duty in mitosis. PMID- 10383304 TI - The Web and conflict of interest. PMID- 10383305 TI - Phytohormone-independent division of tobacco protoplast-derived cells: retractions. PMID- 10383306 TI - The heroin prescribing debate: integrating science and politics. PMID- 10383307 TI - Instruction, selection, or tampering with the odds? PMID- 10383308 TI - Seeking ligands for lonely orphan receptors. PMID- 10383309 TI - Techview: biochemistry. Biomolecule mass spectrometry. PMID- 10383311 TI - Microbes immunity, and disease. Introduction. Microbe management. PMID- 10383310 TI - Dracula's wish. PMID- 10383312 TI - A symphony of bacterial voices. PMID- 10383313 TI - Is it time to uproot the tree of life? PMID- 10383314 TI - Human Genome Project. Sequencers endorse plan for a draft in 1 year. PMID- 10383315 TI - A new look at the martian landscape. PMID- 10383316 TI - Genetically modified food. Britain struggles to turn anti-GM tide. PMID- 10383317 TI - Imaging living cells the friendly way. PMID- 10383318 TI - NIH proposes rules for materials exchange. PMID- 10383319 TI - Community divides over push for bigger budget. PMID- 10383320 TI - Scientific cross-claims fly in continuing beef war. PMID- 10383321 TI - More regulation of rodents. PMID- 10383322 TI - UC-Genentech trial. PMID- 10383323 TI - UC-Genentech trial. PMID- 10383324 TI - A rational approach to labeling biotech-derived foods. PMID- 10383325 TI - Hole-istic medicine. PMID- 10383326 TI - TB vaccines: global solutions for global problems. PMID- 10383327 TI - Gertrude Belle Elion (1918-1999). PMID- 10383329 TI - 40 steps to a chemical synthesis summit. PMID- 10383328 TI - Organ transplants. New drug blocks rejection in monkeys. PMID- 10383330 TI - Behavioral genetics. Fickle mice highlight test problems. PMID- 10383331 TI - Expert testimony. Project offers judges neutral science advice. PMID- 10383332 TI - New leads to cancer, arthritis therapies. PMID- 10383333 TI - European researchers grapple with animal rights. PMID- 10383334 TI - New clues to why size equals destiny. PMID- 10383335 TI - Confronting the HIV pandemic. PMID- 10383336 TI - Space-grown neuraminidase crystals. PMID- 10383337 TI - Uncontrolled release of harmful microorganisms. PMID- 10383338 TI - Tissue engineering. PMID- 10383339 TI - Testing of water for arsenic in Bangladesh. PMID- 10383340 TI - Limits of scientific growth. PMID- 10383341 TI - Life in ice-covered oceans. PMID- 10383342 TI - Selfish sentinels. PMID- 10383343 TI - Nota bene: biomedicine. Pain-killer genes. PMID- 10383344 TI - Genetic control of branching morphogenesis. AB - The genetic programs that direct formation of the treelike branching structures of two animal organs have begun to be elucidated. In both the developing Drosophila tracheal (respiratory) system and mammalian lung, a fibroblast growth factor (FGF) signaling pathway is reiteratively used to pattern successive rounds of branching. The initial pattern of signaling appears to be established by early, more global embryonic patterning systems. The FGF pathway is then modified at each stage of branching by genetic feedback controls and other signals to give distinct branching outcomes. The reiterative use of a signaling pathway by both insects and mammals suggests a general scheme for patterning branching morphogenesis. PMID- 10383345 TI - Genes for ageing? Keystone Symposium: Ageing: Genetic and Environmental Influences on Life Span, Durango, Colorado, USA, 2-7 February 1999. PMID- 10383346 TI - Mouse models for cancer at center stage. AACR special meeting: Cancer Biology and the Mutant Mouse: New Methods, New Models, New Insights, Keystone Colorado, USA, 31 January-5 February 1999. PMID- 10383347 TI - The role of epidemiology in determining when evidence is sufficient to support nutrition recommendations. Proceedings of a workshop. Washington DC, USA. October 7-8, 1997. PMID- 10383348 TI - Low calorie intake in dialysis patients: an alternative explanation. PMID- 10383349 TI - Proceedings of the 18th meeting of the Japanese Society for Biomedical Research on Alcohol. Tokyo, Japan, March 6-7, 1998. PMID- 10383350 TI - Toward evidence-based medical statistics. 2: The Bayes factor. AB - Bayesian inference is usually presented as a method for determining how scientific belief should be modified by data. Although Bayesian methodology has been one of the most active areas of statistical development in the past 20 years, medical researchers have been reluctant to embrace what they perceive as a subjective approach to data analysis. It is little understood that Bayesian methods have a data-based core, which can be used as a calculus of evidence. This core is the Bayes factor, which in its simplest form is also called a likelihood ratio. The minimum Bayes factor is objective and can be used in lieu of the P value as a measure of the evidential strength. Unlike P values, Bayes factors have a sound theoretical foundation and an interpretation that allows their use in both inference and decision making. Bayes factors show that P values greatly overstate the evidence against the null hypothesis. Most important, Bayes factors require the addition of background knowledge to be transformed into inferences- probabilities that a given conclusion is right or wrong. They make the distinction clear between experimental evidence and inferential conclusions while providing a framework in which to combine prior with current evidence. PMID- 10383351 TI - An association for patient-oriented research. AB - Physician-scientists, who study the pathogenesis of disease by using both bedside observations and modern laboratory techniques, are decreasing in number. Ideally, in any profession, some members are devoted to developing its scholarly basis; in contrast, technologists perform activities derived from the work of scientists or scholars. Therefore, the decreasing number of physician-scientists may cause medicine to become a technology rather than a profession. This atrophy in patient oriented research is caused in part by the current state of laboratory science: Discoveries that are not generated from clinical observations often elucidate many aspects of disease. Furthermore, economic necessities have impelled research oriented physicians to choose between delivery of health care and performance of basic laboratory science. This paper discusses the continuing need for detailed observation of human disease as a driving force in the development of biomedical science, which combines clinical and laboratory observations. To further the development of this field, the Association for Patient-Oriented Research has been founded. This association will be a new forum in which physician-scientists can present their work and encourage other physicians to join in the research endeavor. PMID- 10383352 TI - Will the real hemochromatosis please stand up? PMID- 10383353 TI - Standing statistics right side up. PMID- 10383354 TI - Same old seventeen-dollar lamps. PMID- 10383355 TI - The ritual. PMID- 10383356 TI - Subcutaneous heparin for deep venous thrombosis. PMID- 10383357 TI - Ventilator-associated pneumonia in critically ill patients. PMID- 10383358 TI - Increased serum lipoprotein(a) levels in patients with early renal failure. PMID- 10383359 TI - Increased serum lipoprotein(a) levels in patients with early renal failure. PMID- 10383360 TI - Recurrent human granulocytic ehrlichiosis and Lyme disease. PMID- 10383361 TI - Ferrous sulfate tolerance test: a case report. PMID- 10383362 TI - Weighing the alternatives. PMID- 10383363 TI - Nutrition and policy. 3: Food industry response to nutritional standards. PMID- 10383364 TI - Have oxygen, will travel. PMID- 10383365 TI - HFE genotype in patients with hemochromatosis and other liver diseases. AB - BACKGROUND: Hereditary hemochromatosis is a common inherited disorder of iron metabolism. The gene HFE, which contains two missense mutations (C282Y and H63D), was recently identified. OBJECTIVE: To determine how HFE genotyping for the C282Y and H63D mutations contributes to the diagnosis of hemochromatosis and to determine the prevalence of HFE mutations in a group of patients with liver disease. DESIGN: Cross-sectional study. SETTING: Academic medical center. PATIENTS: 66 patients with hereditary hemochromatosis and 132 referred patients with other liver diseases. MEASUREMENTS: At initial diagnosis, fasting transferrin saturation, ferritin level, routine chemistry panel, and complete blood count were determined. Percutaneous liver biopsy was done on all patients for histologic analysis and measurement of hepatic iron concentration and hepatic iron index. HFE genotyping for the C282Y and H63D mutations was done on all patients by using genomic DNA samples. RESULTS: Of the 66 patients with hemochromatosis diagnosed on the basis of serum iron studies and liver biopsy findings, 60 (91%) were C282Y homozygotes, 2 (3%) were compound heterozygotes, 1 (1.5%) was a C282Y heterozygote, 2 (3%) were H63D heterozygotes, and 1 (1.5%) was negative for both mutations. Of the 132 patients with liver disease, 6 (5%) were C282Y homozygotes, 8 (6%) were compound heterozygotes, 6 (5%) were C282Y heterozygotes, 5 (4%) were H63D homozygotes, 20 (15%) were H63D heterozygotes, and 87 (66%) were negative for both mutations. All 66 C282Y homozygotes had an elevated hepatic iron concentration, and 65 of the 66 patients (98%) had a transferrin saturation of at least 45%. Ten of the 66 patients (15% [95% CI, 7.5% to 26%]) had a hepatic iron index less than 1.9 mmol/kg per year; hemochromatosis was not suspected in 6 of the 10 patients before genotyping. Cirrhosis or substantial hepatic fibrosis was not seen in any (0% [CI, 0% to 18%]) of the 19 patients younger than 40 years of age who were homozygous for the C282Y mutation. CONCLUSIONS: All 66 patients homozygous for the C282Y mutation of HFE had an elevated hepatic iron concentration, but approximately 15% of these patients did not meet a previous diagnostic criterion for hemochromatosis (hepatic iron index > 1.9 mmol/kg per year). Determination of HFE genotype is clinically useful in patients with liver disease and suspected iron overload and may lead to identification of otherwise unsuspected C282Y homozygotes. PMID- 10383366 TI - Relation of consumption of vitamin E, vitamin C, and carotenoids to risk for stroke among men in the United States. AB - BACKGROUND: Antioxidants increase the resistance of low-density lipoprotein to oxidation and may thereby reduce risk for atherosclerosis. OBJECTIVE: To determine whether intake of vitamin E, vitamin C, or carotenoids predict risk for total or ischemic stroke. DESIGN: Prospective observational study. SETTING: The Health Professionals Follow-up Study. PARTICIPANTS: 43,738 men 40 to 75 years of age who did not have cardiovascular disease or diabetes. MEASUREMENTS: Repeated and validated dietary assessments were done by using a self-administered 131-item food-frequency questionnaire, which included questions on dose and duration of vitamin supplement use. The follow-up period was 8 years. RESULTS: A total of 328 strokes occurred: 210 ischemic, 70 hemorrhagic, and 48 unclassified. After adjustment for age, smoking, hypertension, hypercholesterolemia, body mass index, physical activity, parental history of myocardial infarction, alcohol consumption, and total energy intake, the relative risk for ischemic stroke in the top quintile of vitamin E intake (median, 411 IU/d) compared with the bottom quintile (5.4 IU/d) was 1.18 (95% CI, 0.77 to 1.82). The relative risk for ischemic stroke in the top quintile of vitamin C intake (1167 mg/d) compared with the bottom quintile (95 mg/d) was 1.03 (CI, 0.66 to 1.59). Results for total stroke were similar. Associations of vitamin intake with hemorrhagic stroke were also nonsignificant, but the CIs were wide. Neither dose nor duration of vitamin E or vitamin C supplement use was related to risk for total or ischemic stroke. The relative risk for ischemic stroke was 1.16 (CI, 0.81 to 1.67) in men using 250 IU or more of vitamin E supplementation per day compared with men who used no vitamin E supplements and was 0.93 (CI, 0.60 to 1.45) in men using 700 mg or more of vitamin C supplementation per day compared with men who used no vitamin C supplements. A significant inverse relation between lutein intake and risk for ischemic stroke was seen but was not independent of other dietary factors. CONCLUSIONS: Vitamin E and vitamin C supplements and specific carotenoids did not seem to substantially reduce risk for stroke in this cohort. Modest effects, however, cannot be excluded. PMID- 10383367 TI - A molecular epidemiologic analysis of tuberculosis trends in San Francisco, 1991 1997. AB - BACKGROUND: To decrease tuberculosis case rates and cases due to recent infection (clustered cases) in San Francisco, California, tuberculosis control measures were intensified beginning in 1991 by focusing on prevention of Mycobacterium tuberculosis transmission and on the use of preventive therapy. OBJECTIVE: To describe trends in rates of tuberculosis cases and clustered cases in San Francisco from 1991 through 1997. DESIGN: Population-based study. SETTING: San Francisco, California. PATIENTS: Persons with tuberculosis diagnosed between 1 January 1991 and 31 December 1997. MEASUREMENTS: DNA fingerprinting was performed. During sequential 1-year intervals, changes in annual case rates per 100,000 persons for all cases, clustered cases (cases with M. tuberculosis isolates having identical fingerprint patterns), and cases in specific subgroups with high rates of clustering (persons born in the United States and HIV-infected persons) were examined. RESULTS: Annual tuberculosis case rates peaked at 51.2 cases per 100,000 persons in 1992 and decreased significantly thereafter to 29.8 cases per 100,000 persons in 1997 (P < 0.001). The rate of clustered cases decreased significantly over time in the entire study sample (from 10.4 cases per 100,000 persons in 1991 to 3.8 cases per 100,000 persons in 1997 [P < 0.001]), in persons born in the United States (P < 0.001), and in HIV-infected persons (P = 0.003). CONCLUSIONS: The rates of tuberculosis cases and clustered tuberculosis cases decreased both overall and among persons in high-risk groups. This occurred in a period during which tuberculosis control measures were intensified. PMID- 10383368 TI - Translating clinical trial results into practice: the effect of an AIDS clinical trial on prescribed antiretroviral therapy for HIV-infected pregnant women. AB - BACKGROUND: The success of Pediatric AIDS Clinical Trials Group (PACTG) Protocol 076 in preventing vertical HIV transmission prompted intensive efforts to inform lay-persons and professionals about the trial's results. OBJECTIVE: To explore community responsiveness to these efforts by assessing temporal, maternal, and health care factors associated with prescribed antiretroviral therapy before and after PACTG Protocol 076. DESIGN: Retrospective cohort study. SETTING: New York State Medicaid program. PATIENTS: 2607 HIV-infected women who delivered a living child between January 1993 and September 1996. MEASUREMENTS: Adjusted odds of being prescribed antiretroviral treatment in the second or third trimester for women who delivered in period 1 (during the trial [January 1993 to February 1994]), period 2 (after the trial's end and announcement of the results to publication of the results [March 1994 to November 1994]), and period 3 (after publication of the trial results [December 1994 to September 1996]). RESULTS: The adjusted odds of being prescribed antiretroviral therapy increased 21% per month in period 2 and decreased to 3% per month in period 3. In all time periods, the adjusted odds of being prescribed antiretroviral therapy were at least 60% greater (P < 0.05) for women who were treated at an institution that performed HIV clinical trials, received HIV-focused ambulatory care, or had adequate prenatal care visits. After the trial, women receiving methadone treatment had at least twofold (95% CI, 1.5- to 4.3-fold) greater adjusted odds of being prescribed antiretroviral therapy than women who did not take any illicit drugs. Latin-American women, older women, and women born in the United States had greater adjusted odds (P < 0.05) of being prescribed treatment in period 3. CONCLUSION: Community practice responded rapidly to efforts to disseminate the results of PACTG Protocol 076; however, the absolute increase in prescribed therapy was greatest for women who had adequate prenatal visits or were receiving HIV-focused care, care at a site that performed clinical trials, or methadone therapy. PMID- 10383369 TI - Effects of leisure-time physical activity and ventilatory function on risk for stroke in men: the Reykjavik Study. AB - BACKGROUND: Stroke is a major cause of illness, death, and health expenditures. Leisure-time physical activity may reduce the risk for stroke. OBJECTIVE: To examine the association of leisure-time physical activity and pulmonary function with risk for stroke. DESIGN: Prospective cohort study. SETTING: Reykjavik, Iceland. PARTICIPANTS: 4484 men 45 to 80 years of age followed for a mean (+/-SD) of 10.6 +/- 3.6 years. MEASUREMENTS: Patients underwent physical examination, blood sampling, and spirometry and completed a questionnaire about health and exercise. Computerized hospital records were used to identify strokes, and the Icelandic National Registry was used to identify deaths. RESULTS: New stroke developed in 249 men (5.6%) (hemorrhagic stroke in 44 [18%] and ischemic stroke in 205 [82%]). In a multivariable hazard analysis that controlled for known risk factors for cerebrovascular disease, leisure-time physical activity maintained after 40 years of age was associated with a reduced risk for stroke (relative risk, 0.69 [CI, 0.47 to 1.01] for total stroke and 0.62 [CI, 0.40 to 0.97] for ischemic stroke). Risk for stroke increased with diminished ventilatory function (FVC or FEV1) (relative risk, 1.9 [CI, 1.06 to 3.25] for the lowest compared with the highest quintile). CONCLUSION: Middle-aged men who participate in leisure time physical activity and have good pulmonary function seem to have a lower risk for stroke than men who are not active or have diminished pulmonary function. PMID- 10383371 TI - Toward evidence-based medical statistics. 1: The P value fallacy. AB - An important problem exists in the interpretation of modern medical research data: Biological understanding and previous research play little formal role in the interpretation of quantitative results. This phenomenon is manifest in the discussion sections of research articles and ultimately can affect the reliability of conclusions. The standard statistical approach has created this situation by promoting the illusion that conclusions can be produced with certain "error rates," without consideration of information from outside the experiment. This statistical approach, the key components of which are P values and hypothesis tests, is widely perceived as a mathematically coherent approach to inference. There is little appreciation in the medical community that the methodology is an amalgam of incompatible elements, whose utility for scientific inference has been the subject of intense debate among statisticians for almost 70 years. This article introduces some of the key elements of that debate and traces the appeal and adverse impact of this methodology to the P value fallacy, the mistaken idea that a single number can capture both the long-run outcomes of an experiment and the evidential meaning of a single result. This argument is made as a prelude to the suggestion that another measure of evidence should be used--the Bayes factor, which properly separates issues of long-run behavior from evidential strength and allows the integration of background knowledge with statistical findings. PMID- 10383370 TI - Effect of nasogastric tube size on gastroesophageal reflux and microaspiration in intubated patients. AB - BACKGROUND: Little evidence exists to support the theory that small-bore nasogastric tubes prevent gastroesophageal reflux and microaspiration in intubated patients. OBJECTIVE: To determine whether gastroesophageal reflux and microaspiration in intubated patients can be reduced by the use of a small-bore nasogastric tube. DESIGN: Randomized, two-period crossover trial. SETTING: Respiratory intensive care unit of a university hospital. PATIENTS: 17 patients intubated for more than 72 hours. INTERVENTIONS: Radioactive technetium colloid was instilled in each patient's stomach. Patients were studied with two nasogastric tubes (one tube with a 6.0-mm external bore and one tube with a 2.85 mm external bore) in randomized order; measurements of radioactive counts with the alternate size of nasogastric tube were repeated 72 hours after original measurements were taken. Sequential samples of serum, gastric juice, and pharyngeal and tracheal secretions were obtained. MEASUREMENTS: Comparison of the time course of radioactive counting in all samples (obtained during the use of each nasogastric tube size in each patient). RESULTS: The mean radioactive count of pharyngeal aspirates (P = 0.004) was greater than the baseline count at all time points, as was the cumulative radioactive count of pharyngeal aspirates 17 hours after the first dose of technetium colloid was administered (P = 0.001); however, the count of tracheal aspirates was never greater than the count at baseline. No differences were found between tube types when the time course and cumulative counts of pharyngeal and tracheal samples were compared. CONCLUSION: Small-bore nasogastric tubes in intubated patients do not reduce gastroesophageal reflux or microaspiration. PMID- 10383372 TI - Limitations of cytomegalovirus testing. PMID- 10383374 TI - Increasing resistance of Brucellae to co-trimoxazole. PMID- 10383373 TI - In vitro activities of clarithromycin and azithromycin against clinical isolates of Mycobacterium avium-M. intracellulare. PMID- 10383375 TI - Expression of c-erbB-4/HER4 is regulated in T47D breast carcinoma cells by retinoids and vitamin D3. AB - Nuclear steroid/retinoid and memgbrane c-erbB receptor tyrosine kinase signaling control proliferation and differentiation of mammary epithelial cells. Recently, we reported that retinoic acids are efficient repressors of c-erbB-2 and -3, but not of c-erbB-1 gene expresson. Here we demonstrate that retinoid acid- mediated growth inhibition is accompanied with reduced expression of c-erbB-4/HER4 in T47D breast cancer cells as determined by FACS, Western, and RT-PCR. All-trans and 9 cis retinoic acid reduce c-erbB-4 expression to 30%-60% of control, depending on the concentration. Dexamethasone (Dex) is inactive on D3 (D3), in contrast, acts as a strong inducer, elevation more that twofold at the mRNA, but does not significantly affect cell growth. We concolude that retinoic acids are efficient growth inhibitors and repressors of cell growth and c-erbB-4, whereas D3 represents a highly efficient inducer of c-erbB-4 expression with affecting cell proliferation. PMID- 10383376 TI - Isolation and characterization of low density detergent-insoluble membrane (LD DIM) fraction from sea urchin sperm. AB - The low density detergent-insoluble membrane (LD-DIM) fraction was obtained by a sucrose-density gradient centrifugation from sperm of three sea urchin species, Hemicentrotus pulcherrimus, Strongylocentrotus purpuratus, and Anthocidaris crassispina. These LD-DIM preparations were characterized by enriched glycosphingolipids (GSL) including gangliosides and sulfatide (SLF), having more than 50% of the total amount of GSL present in these sperm. Interestingly, a minor component of H. pulcherrimus sperm (HO3S-->8Neu5Acalpha2-->8Neu5Acalpha2- >6Glcbeta1++ +-->Cer) was shown to be even more enriched in the LD-DIM as revealed by using monoclonal antibody (mAb.3G9) speific to this ganglioside. In addition to the GSL, phosphatidyl-serine (PS) and diacylglcerol (DG) were enriched in the LD-DIM. On the other hand, cholesterol (CL) and sphingomyelin (SM) were not so enriched, which contrasted with the LD-DIM from Madin-Darby canine kidney (MDCK) cells, where CL and SM were reported to be abundant. Because mammalian somatic cell-derived DIMs have been proposed to be associated with functional signal transduction, it seems possible that the ganglioside-enriched LD-DIM in sea urchin sperm can participate in binding to eggs and the subsequent egg activation process. To our knowledge this is the chemical characterization of the LD-DIM fraction of a gametic cell. PMID- 10383377 TI - A randomized evaluation of early revascularization to treat shock complicating acute myocardial infarction. The (Swiss) Multicenter Trial of Angioplasty for Shock-(S)MASH. AB - AIM: To test whether emergency revascularization improves survival in patients with acute myocardial infarction and shock. METHODS AND RESULTS: Patients with acute myocardial infarction and early shock were randomized either to undergo emergency angiography, followed immediately by revascularization when indicated, or to receive initial medical management. In five of the nine participating centres, patients with shock but not randomized were entered in a registry. Only 55 patients could be randomized. Of the 32 patients in the invasive group, 30 (94%) underwent early angiography, 27 (84%) PTCA, and one (4%) CABG. Twenty-two (69%) died within 30 days in the invasive group vs 18/23 (78%) in the medically managed group (ns, RR=0.88, 95% confidence interval 0.6-1.2). Among the registry patients, 24/51 were excluded from randomization solely because of patient or physician preference for the invasive approach: 23 (96%) of them underwent emergency angiography, 21 (88%) PTCA, and 12 (50%) died within 30 days. Among the remaining registry patients (n=27) only nine (33%) underwent early angiography, nine (33%) PTCA and 20 (74%) died. CONCLUSION: We failed to demonstrate that emergency PTCA significantly improves survival in patients with acute myocardial infarction and early cardiogenic shock. Because the study was stopped prematurely, due to an insufficient patient inclusion rate, a clinically meaningful benefit of early reperfusion may have been missed. PMID- 10383378 TI - The biology of the small leucine-rich proteoglycans. Functional network of interactive proteins. PMID- 10383379 TI - TAF25p, a non-histone-like subunit of TFIID and SAGA complexes, is essential for total mRNA gene transcription in vivo. AB - We demonstrate, utilizing a temperature conditional mutant allele of the gene encoding TAF25p, that this non-histone-like TBP-associated factor, which is shared between the TFIID and SAGA complexes, is required for bulk mRNA gene transcription by RNA polymerase II in vivo. Immunoblotting experiments indicate that at the restrictive temperature, inactivation of TAF25p function results in a reduction of the levels of numerous TFIID and SAGA subunits, indicating its loss of function, like the histone-like TAFs, causes degradation of the constituents of these two multisubunit complexes. These data suggest that TAF25p plays a key structural role in maintaining TFIID and SAGA complex integrity. This is the first demonstration that a non-histone-like TAF is required for continuous, high level RNA polymerase II-mediated mRNA gene transcription in living yeast cells. PMID- 10383380 TI - Mutagenesis identifies new signals for beta-amyloid precursor protein endocytosis, turnover, and the generation of secreted fragments, including Abeta42. AB - It has long been assumed that the C-terminal motif, NPXY, is the internalization signal for beta-amyloid precursor protein (APP) and that the NPXY tyrosine (Tyr743 by APP751 numbering, Tyr682 in APP695) is required for APP endocytosis. To evaluate this tenet and to identify the specific amino acids subserving APP endocytosis, we mutated all tyrosines in the APP cytoplasmic domain and amino acids within the sequence GYENPTY (amino acids 737-743). Stable cell lines expressing these mutations were assessed for APP endocytosis, secretion, and turnover. Normal APP endocytosis was observed for cells expressing Y709A, G737A, and Y743A mutations. However, Y738A, N740A, and P741A or the double mutation of Y738A/P741A significantly impaired APP internalization to a level similar to that observed for cells lacking nearly the entire APP cytoplasmic domain (DeltaC), arguing that the dominant signal for APP endocytosis is the tetrapeptide YENP. Although not an APP internalization signal, Tyr743 regulates rapid APP turnover because half-life increased by 50% with the Y743A mutation alone. Secretion of the APP-derived proteolytic fragment, Abeta, was tightly correlated with APP internalization, such that Abeta secretion was unchanged for cells having normal APP endocytosis but significantly decreased for endocytosis-deficient cell lines. Remarkably, secretion of the Abeta42 isoform was also reduced in parallel with endocytosis from internalization-deficient cell lines, suggesting an important role for APP endocytosis in the secretion of this highly pathogenic Abeta species. PMID- 10383381 TI - Activated Ha-ras induces apoptosis by association with phosphorylated Bcl-2 in a mitogen-activated protein kinase-independent manner. AB - Serum deprivation of Ha-ras-transformed brown adipocyte cell line resulted in a dramatic apoptotic cell death, as detected either by DNA laddering or by an increase in the percentage of hypodiploid cells or by nuclei condensation and fragmentation, as compared with immortalized cell line or primary fetal brown adipocytes. Moreover, transient transfection of immortalized brown adipocytes with a constitutively active ras gene (Ha-raslys12) mimics the high rate of apoptosis detected in the transformed cell line. On the other hand, transient transfection of the dominant-negative construct of raf-1 rescued serum-deprived Ha-ras-transformed brown adipocytes from apoptosis, decreasing the percentage of hypodiploid cells, the external display of phosphatidylserine, and the DNA laddering. However, inhibition of mitogen-activated protein kinase with PD098059 did not preclude apoptosis and in fact increased the rate of apoptosis observed in serum-deprived Ha-ras-transformed cells, indicating that the Ras/Raf-1 pathway induced apoptosis throughout a mitogen-activated protein kinase kinase 1 (MEK-1) independent pathway. Furthermore, apoptosis in Ha-ras-transformed brown adipocytes is concurrent with an up-regulation in the expression of the pro apoptotic protein Bcl-xS, the expression of the anti-apoptotic protein Bcl-2 being down-regulated. Finally, an association of Ras and Raf with phosphorylated Bcl-2 protein was demonstrated in immunoprecipitates from apoptotic cells. Thus, we propose a mechanism of apoptosis in Ha-ras-transformed adipocytes under serum deprivation involving Raf-1 association with phosphorylated Bcl-2, down regulation of Bcl-2 expression, and up-regulation of Bcl-xS expression. PMID- 10383382 TI - Competitive, reversible inhibition of cytosolic phospholipase A2 at the lipid water interface by choline derivatives that partially partition into the phospholipid bilayer. AB - Cytosolic phospholipase A2 (cPLA2) catalyzes the selective release of arachidonic acid from the sn-2 position of phospholipids and is believed to play a key cellular role in the generation of arachidonic acid. When assaying the human recombinant cPLA2 using membranes isolated from [3H]arachidonate-labeled U937 cells as substrate, 2-(2'-benzyl-4-chlorophenoxy)ethyl-dimethyl-n-octadecyl ammonium chloride (compound 1) was found to inhibit the enzyme in a dose dependent manner (IC50 = 5 microM). It was over 70 times more selective for the cPLA2 as compared with the human nonpancreatic secreted phospholipase A2, and it did not inhibit other phospholipases. Additionally, it inhibited arachidonate production in N-formyl-methionyl-leucyl-phenylalanine-stimulated U937 cells. To further characterize the mechanism of inhibition, an assay in which the enzyme is bound to vesicles of 1,2-dimyristoyl-sn -glycero-3-phosphomethanol containing 6 10 mol % of 1-palmitoyl-2-[1-14C]arachidonoyl-sn-glycero-3-phosphocholine was employed. With this substrate system, the dose-dependent inhibition could be defined by kinetic equations describing competitive inhibition at the lipid-water interface. The apparent equilibrium dissociation constant for the inhibitor bound to the enzyme at the interface (KI*app) was determined to be 0.097 +/- 0.032 mol % versus an apparent dissociation constant for the arachidonate-containing phospholipid of 0.3 +/- 0.1 mol %. Thus, compound 1 represents a novel structural class of inhibitor of cPLA2 that partitions into the phospholipid bilayer and competes with the phospholipid substrate for the active site. Shorter n-alkyl chained (C-4, C-6, C-8) derivatives of compound 1 were shown to have even smaller KI*app values. However, these short-chained analogs were less potent in terms of bulk inhibitor concentration needed for inhibition when using the [3H]arachidonate-labeled U937 membranes as substrate. This discrepancy was reconciled by showing that these shorter-chained analogs did not partition into the [3H]arachidonate-labeled U937 membranes as effectively as compound 1. The implications for in vivo efficacy that result from these findings are discussed. PMID- 10383383 TI - Salivary histatin 5 induces non-lytic release of ATP from Candida albicans leading to cell death. AB - Salivary histatins are potent in vitro antifungal proteins and have promise as therapeutic agents against oral candidiasis. We performed pharmacological studies directed at understanding the biochemical basis of Hst 5 candidacidal activity. Three inhibitors of mitochondrial metabolism: carbonyl cyanide p chlorophenylhydrazone, dinitrophenol, and azide inhibited Hst 5 killing of Candida albicans, while not inhibiting cellular ATP production. In contrast, Hst 5 caused a drastic reduction of C. albicans intracellular ATP content, which was a result of an efflux of ATP. Carbonyl cyanide p-chlorophenylhydrazone, dinitrophenol, and azide inhibited Hst 5-induced ATP efflux, thus establishing a correlation between ATP release and cell killing. Furthermore, C. albicans cells were respiring and had polarized membranes at least 80 min after ATP release, thus implying a non-lytic exit of cellular ATP in response to Hst 5. Based on evidence that transmembrane ATP efflux can occur in the absence of cytolysis through a channel-like pathway and that released ATP can act as a cytotoxic mediator by binding to membrane purinergic receptors, we evaluated whether extracellular ATP released by Hst 5 may have further functional role in cell killing. Consistent with this hypothesis, purinergic agonists BzATP and adenosine 5'O-(thiotriphosphate) induced loss of C. albicans cell viability and purinergic antagonists prevented Hst 5 killing. PMID- 10383384 TI - Functional effect of deletion and mutation of the Escherichia coli ribosomal RNA and tRNA pseudouridine synthase RluA. AB - The Escherichia coli gene rluA, coding for the pseudouridine synthase RluA that forms 23 S rRNA pseudouridine 746 and tRNA pseudouridine 32, was deleted in strains MG1655 and BL21/DE3. The rluA deletion mutant failed to form either 23 S RNA pseudouridine 746 or tRNA pseudouridine 32. Replacement of rluA in trans on a rescue plasmid restored both pseudouridines. Therefore, RluA is the sole protein responsible for the in vivo formation of 23 S RNA pseudouridine 746 and tRNA pseudouridine 32. Plasmid rescue of both rluA- strains using an rluA gene carrying asparagine or threonine replacements for the highly conserved aspartate 64 demonstrated that neither mutant could form 23 S RNA pseudouridine 746 or tRNA pseudouridine 32 in vivo, showing that this conserved aspartate is essential for enzyme-catalyzed formation of both pseudouridines. In vitro assays using overexpressed wild-type and mutant synthases confirmed that only the wild-type protein was active despite the overexpression of wild-type and mutant synthases in approximately equal amounts. There was no difference in exponential growth rate between wild-type and MG1655(rluA-) either in rich or minimal medium at 24, 37, or 42 degrees C, but when both strains were grown together, a strong selection against the deletion strain was observed. PMID- 10383385 TI - Adenylyl cyclase, a coincidence detector for nitric oxide. AB - Nitric oxide (NO) donors inhibit hormone- and forskolin-stimulated adenylyl cyclase activity in purified plasma membrane preparations from N18TG2 neuroblastoma cells. Northern blot analyses indicate that the predominant isoform of adenylyl cyclase in N18TG2 cells is the type VI. Our experiments eliminate all the known regulatory proteins for this isoform as possible targets of NO. NO decreases the Vmax of the enzyme without altering the Km for ATP. Occupancy of the substrate-binding site protects the enzyme from the inhibitory effects of NO, suggesting that the conformation of the enzyme determines its sensitivity. The inhibition is reversed by reducing agents, implicating a Cys residue(s) as the target for nitric oxide and an S-nitrosylation as the underlying modification. These findings implicate NO as a novel cellular regulator of the type VI isoform of adenylyl cyclase. PMID- 10383386 TI - Synaptogyrins regulate Ca2+-dependent exocytosis in PC12 cells. AB - Synaptogyrins constitute a family of synaptic vesicle proteins of unknown function. With the full-length structure of a new brain synaptogyrin isoform, we now show that the synaptogyrin family in vertebrates includes two neuronal and one ubiquitous isoform. All of these synaptogyrins are composed of a short conserved N-terminal cytoplasmic sequence, four homologous transmembrane regions, and a variable cytoplasmic C-terminal tail that is tyrosine-phosphorylated. The localization, abundance, and conservation of synaptogyrins suggest a function in exocytosis. To test this, we employed a secretion assay in PC12 cells expressing transfected human growth hormone (hGH) as a reporter protein. When Ca2+-dependent hGH secretion from PC12 cells was triggered by high K+ or alpha-latrotoxin, co transfection of all synaptogyrins with hGH inhibited hGH exocytosis as strongly as co-transfection of tetanus toxin light chain. Synaptophysin I, which is distantly related to synaptogyrins, was also inhibitory but less active. Inhibition was independent of the amount of hGH expressed but correlated with the amount of synaptogyrin transfected. Inhibition of exocytosis was not observed with several other synaptic proteins, suggesting specificity. Analysis of the regions of synaptogyrin required for inhibition revealed that the conserved N terminal domain of synaptogyrin is essential for inhibition, whereas the long C terminal cytoplasmic tail is largely dispensable. Our results suggest that synaptogyrins are conserved components of the exocytotic apparatus, which function as regulators of Ca2+-dependent exocytosis. PMID- 10383387 TI - Extracellular cysteines of CCR5 are required for chemokine binding, but dispensable for HIV-1 coreceptor activity. AB - CCR5 is the major coreceptor for macrophage-tropic human immunodeficiency virus type I (HIV-1). For most G-protein-coupled receptors that have been tested so far, the disulfide bonds linking together the extracellular loops (ECL) are required for maintaining the structural integrity necessary for ligand binding and receptor activation. A natural mutation affecting Cys20, which is thought to form a disulfide bond with Cys269, has been described in various human populations, although the consequences of this mutation for CCR5 function are not known. Using site-directed mutagenesis, we mutated the four extracellular cysteines of CCR5 singly or in combination to investigate their role in maintaining the structural conformation of the receptor, its ligand binding and signal transduction properties, and its ability to function as a viral coreceptor. Alanine substitution of any single Cys residue reduced surface expression levels by 40-70%. However, mutation of Cys101 or Cys178, predicted to link ECL1 and ECL2 of the receptor, abolished recognition of CCR5 by a panel of conformation sensitive anti-CCR5 antibodies. The effects of the mutations on receptor expression and conformation were partially temperature-sensitive, with partial restoration of receptor expression and conformation achieved by incubating cells at 32 degrees C. All cysteine mutants were unable to bind detectable levels of MIP-1beta, and did not respond functionally to CCR5 agonists. Surprisingly, all cysteine mutants did support infection by R5 strains of HIV, though at reduced levels. These results indicate that both disulfide bonds of CCR5 are necessary for maintaining the structural integrity of the receptor necessary for ligand binding and signaling. Env binding and the mechanisms of HIV entry appear much less sensitive to alterations of CCR5 conformation. PMID- 10383388 TI - Schmid's metaphyseal chondrodysplasia mutations interfere with folding of the C terminal domain of human collagen X expressed in Escherichia coli. AB - Human collagen X contains a highly conserved 161-amino acid C-terminal non-triple helical domain that is homologous to the C-terminal domain of collagen VIII and to the C1q module of the human C1 enzyme. We have expressed this domain (residues 545-680) in Escherichia coli as a glutathione S-transferase fusion protein. The purified fusion protein trimerizes spontaneously in vitro, and after thrombin cleavage, the purified C-terminal domain trimer (46.2 kDa) is extremely stable and trypsin-resistant. Mutations within the C-terminal domain have been observed in patients with Schmid's metaphyseal chondrodysplasia (SMCD). Some of these mutations (Y598D, G618V, W651X, or H669X; X is the stop codon) were constructed by site-directed mutagenesis. Each mutation had identical consequences regarding the fusion protein: 1) absence of trimeric formation, 2) copurification of the approximately 60-kDa GroEL chaperone protein, and 3) sensitivity of the monomeric fusion protein to trypsin digestion. These results show that the C-terminal domain of collagen X is sufficient to produce a very stable and compact trimer in the absence of collagen Gly-X-Y repeats. Moreover, mutations causing SMCD interfere in this system with the correct folding of the C-terminal domain. The existence of a similar mechanism in chondrocytes might explain the relative homogeneity of phenotypes in SMCD despite the diversity of mutations. PMID- 10383389 TI - Oxidative cross-linking of ApoB100 and hemoglobin results in low density lipoprotein modification in blood. Relevance to atherogenesis caused by hemodialysis. AB - Human blood contains a form of minimally modified low density lipoprotein (LDL), termed LDL-, whose origin remains unknown. Exploring the mechanism of formation, we found that LDL- can be produced in plasma in the absence of oxygen following LDL incubation with oxidized hemoglobin species. A high degree of apolipoprotein B100 modification results from covalent association of hemoglobin with LDL involving dityrosine formation but not due to the malonaldehyde epitope formation. This was evidenced by the cross-reactivity of oxidized LDL with antibodies against hemoglobin that was accompanied by a 60-fold increase in dityrosine levels. In this study we found significantly higher LDL- levels in the blood of hemodialysis patients, perhaps contributing to their greatly increased risk of atherosclerosis. The mechanism of LDL- formation was studied during ex vivo blood circulation using a model system resembling clinical hemodialysis in terms of the induction of inflammatory responses. This circulation increased free hemoglobin and LDL- levels compared with non-circulated blood without appreciable lipid peroxidation. Pronounced increases in LDL- were found also during circulation of plasma supplemented with nanomolar hemoglobin levels. The increase in dityrosine content and presence of heme in LDL after blood circulation suggest that LDL is modified, in part, by hemoglobin-LDL conjugates containing heme. Thus, hemoglobin-mediated reactions leading to LDL oxidation in plasma can account for high LDL- levels in hemodialysis patients. PMID- 10383390 TI - All-trans-retinoic acid-mediated growth inhibition involves inhibition of human kinesin-related protein HsEg5. AB - In this study we used differential display reverse transcription-polymerase chain reaction to search for differentially expressed all-trans-retinoic acid (ATRA) responsive genes in pancreatic carcinoma cells. We identified the kinesin-related protein HsEg5, which plays an essential role in spindle assembly and spindle function during mitosis, as a novel molecule involved in ATRA-mediated growth inhibition. Using Northern and Western blot analysis we demonstrated that ATRA significantly inhibits HsEg5 expression in various pancreatic carcinoma cell lines as well as in HaCat keratinocytes. Inhibition of HsEg5 expression by ATRA occurs at the posttranscriptional level. As a consequence, tumor cells synchronized in S-phase revealed a retarded progression through G2/M phase of the cell cycle indicating that HsEg5 inhibition results in a delayed progression through mitosis. Furthermore, a significant decrease of HsEg5 protein expression achieved by antisense transfection revealed a significant growth inhibition compared with control cells. Therefore, HsEg5 represents a novel molecule involved in ATRA-mediated growth inhibition, suggesting that vitamin A derivatives can interact with the bipolar spindle apparatus during mitosis. PMID- 10383391 TI - Hyperphosphorylation of the retinoid X receptor alpha by activated c-Jun NH2 terminal kinases. AB - The nuclear receptor mouse retinoid X receptor alpha (mRXRalpha) was shown to be constitutively phosphorylated in its NH2-terminal A/B region, which contains potential phosphorylation sites for proline-directed Ser/Thr kinases. Mutants for each putative site were generated and overexpressed in transfected COS-1 cells. Constitutively phosphorylated residues identified by tryptic phosphopeptide mapping included serine 22 located in the A1 region that is specific to the RXRalpha1 isoform. Overexpression and UV activation of the stress-activated kinases, c-Jun NH2-terminal kinases 1 and 2 (JNK1 and JNK2), hyperphosphorylated RXRalpha, resulting in a marked decrease in its electrophoretic mobility. This inducible hyperphosphorylation involved three residues (serines 61 and 75 and threonine 87) in the B region of RXRalpha and one residue (serine 265) in the ligand binding domain (E region). Binding assays performed in vitro with purified recombinant proteins demonstrated that JNKs did not interact with RXRalpha but bound to its heterodimeric partners, retinoic acid receptors alpha and gamma (RARalpha and RARgamma). Hyperphosphorylation by JNKs did not affect the transactivation properties of either RXRalpha homodimers or RXRalpha/RARalpha heterodimers in transfected cultured cells. PMID- 10383392 TI - Inactivated pbp4 in highly glycopeptide-resistant laboratory mutants of Staphylococcus aureus. AB - Both vancomycin- and teicoplanin-resistant laboratory mutants of Staphylococcus aureus produce peptidoglycans of altered composition in which the proportion of highly cross-linked muropeptide species is drastically reduced with a parallel increase in the representation of muropeptide monomers and dimers (Sieradzki, K., and Tomasz, A. (1997) J. Bacteriol. 179, 2557-2566; and Sieradzki, K. , and Tomasz, A. (1998) Microb. Drug Resist. 4, 159-168). We now report that the distorted peptidoglycan composition is related to defects in penicillin-binding protein 4 (PBP4); no PBP4 was detectable by the fluorographic assay in membrane preparations from the mutants, and comparison of the sequence of pbp4 amplified from the mutants indicated disruption of the gene by two types of abnormalities, a 17-amino acid long duplication starting at position 305 of the pbp4 gene was detected in the vancomycin-resistant mutant, and a stop codon was found to be introduced into the pbp4 KTG motif at position 261 in the mutant selected for teicoplanin resistance. Additional common patterns of disturbances in the peptidoglycan metabolism of the mutants are indicated by the increased sensitivity of mutant cell walls to the M1 muramidase and decreased sensitivity to lysostaphin, which is a reversal of the susceptibility pattern of the parental cell walls. Furthermore, the results of high performance liquid chromatography analysis of lysostaphin digests of peptidoglycan suggest an increase in the average chain length of the glycan strands in the peptidoglycan of the glycopeptide-resistant mutants. The increased molar proportion of muropeptide monomers in the cell wall of the glycopeptide-resistant mutants should provide binding sites for the "capture" of vancomycin and teicoplanin molecules, which may be part of the mechanism of glycopeptide resistance in S. aureus. PMID- 10383393 TI - Regulation of Hsp27 oligomerization, chaperone function, and protective activity against oxidative stress/tumor necrosis factor alpha by phosphorylation. AB - The small heat shock proteins (sHsps) from human (Hsp27) and mouse (Hsp25) form large oligomers which can act as molecular chaperones in vitro and protect cells from heat shock and oxidative stress when overexpressed. In addition, mammalian sHsps are rapidly phosphorylated by MAPKAP kinase 2/3 at two or three serine residues in response to various extracellular stresses. Here we analyze the effect of sHsp phosphorylation on its quaternary structure, chaperone function, and protection against oxidative stress. We show that in vitro phosphorylation of recombinant sHsp as well as molecular mimicry of Hsp27 phosphorylation lead to a significant decrease of the oligomeric size. We demonstrate that both phosphorylated sHsps and the triple mutant Hsp27-S15D,S78D,S82D show significantly decreased abilities to act as molecular chaperones suppressing thermal denaturation and facilitating refolding of citrate synthase in vitro. In parallel, Hsp27 and its mutants were analyzed for their ability to confer resistance against oxidative stress when overexpressed in L929 and 13.S.1.24 cells. While wild type Hsp27 confers resistance, the triple mutant S15D,S78D,S82D cannot protect against oxidative stress effectively. These data indicate that large oligomers of sHsps are necessary for chaperone action and resistance against oxidative stress whereas phosphorylation down-regulates these activities by dissociation of sHsp complexes to tetramers. PMID- 10383394 TI - CD44, a cell surface chondroitin sulfate proteoglycan, mediates binding of interferon-gamma and some of its biological effects on human vascular smooth muscle cells. AB - Several cytokines and growth factors act on cells after their association with the glycosaminoglycan (GAG) moiety of cell surface proteoglycans (PGs). Interferon-gamma (IFN-gamma) binds to GAG; however, the relevance of this interaction for the biological activity of IFN-gamma on human cells remains to be established. Human arterial smooth muscle cells (HASMC), the main cells synthesizing PG in the vascular wall, respond markedly to IFN-gamma. We found that treatment of HASMC with chondroitinase ABC, an enzyme that degrades chondroitin sulfate GAG, reduced IFN-gamma binding by more than 50%. This treatment increased the affinity of 125I-IFN-gamma for cells from a Kd value of about 93 nM to a Kd value of about 33 nM. However, the total binding was reduced from 9. 3 +/- 0.77 pmol/microg to 3.0 +/- 0.23 pmol/mg (n = 4). Interestingly, pretreatment with chondroitinase ABC reduced significantly the cellular response toward IFN-gamma. The interaction of IFN-gamma with chondroitin sulfate GAG was confirmed by affinity chromatography of isolated cell-associated 35S-, 3H-labeled PG on a column with immobilized IFN-gamma. The cell-associated PG that binds to IFN-gamma was a chondroitin sulfate PG (CSPG). This CSPG had a core protein of approximately 110 kDa that was recognized by anti-CD44 antibodies on Western blots. High molecular weight complexes between IFN-gamma and chondroitin 6 sulfate were observed in gel exclusion chromatography. Additions of chondroitin 6 sulfate to cultured HASMC antagonized the antiproliferative effect and expression of major histocompatibility complex II antigens induced by IFN-gamma. These results indicate that IFN-gamma binds with low affinity to the chondroitin sulfate GAG moiety of the cell surface CSPG receptor CD44. This interaction may increase the local concentration of IFN-gamma at the cell surface, thus facilitating its binding to high affinity receptors and modulating the ability of IFN-gamma to signal a cellular response. PMID- 10383395 TI - Mechanism of endoplasmic reticulum retention of mutant vasopressin precursor caused by a signal peptide truncation associated with diabetes insipidus. AB - Autosomal dominant neurohypophyseal diabetes insipidus is caused by mutations in the gene encoding the vasopressin precursor protein, prepro-vasopressin neurophysin II. We analyzed the molecular consequences of a mutation (DeltaG227) recently identified in a Swiss kindred that destroys the translation initiation codon. In COS-7 cells transfected with the mutant cDNA, translation was found to initiate at an alternative ATG, producing a truncated signal sequence that was functional for targeting and translocation but was not cleaved by signal peptidase. The mutant precursor was completely retained within the endoplasmic reticulum. The uncleaved signal did not affect folding of the neurophysin portion of the precursor, as determined by its protease resistance. However, formation of disulfide-linked aggregates indicated that it interfered with the formation of the disulfide bond in vasopressin, most likely by blocking its insertion into the hormone binding site of neurophysin. Preventing disulfide formation in the vasopressin nonapeptide by mutation of cysteine 6 to serine was shown to be sufficient to cause aggregation and retention. These results indicate that the DeltaG227 mutation induces translation of a truncated signal sequence that cannot be cleaved but prevents correct folding and oxidation of vasopressin, thereby causing precursor aggregation and retention in the endoplasmic reticulum. PMID- 10383396 TI - Inositol 1,3,4-trisphosphate acts in vivo as a specific regulator of cellular signaling by inositol 3,4,5,6-tetrakisphosphate. AB - Ca2+-activated Cl- channels are inhibited by inositol 3,4,5, 6-tetrakisphosphate (Ins(3,4,5,6)P4) (Xie, W., Kaetzel, M. A., Bruzik, K. S., Dedman, J. R., Shears, S. B., and Nelson, D. J. (1996) J. Biol. Chem. 271, 14092-14097), a novel second messenger that is formed after stimulus-dependent activation of phospholipase C (PLC). In this study, we show that inositol 1,3,4-trisphosphate (Ins(1,3,4)P3) is the specific signal that ties increased cellular levels of Ins(3,4,5,6)P4 to changes in PLC activity. We first demonstrated that Ins(1,3,4)P3 inhibited Ins(3,4,5,6)P4 1-kinase activity that was either (i) in lysates of AR4-2J pancreatoma cells or (ii) purified 22,500-fold (yield = 13%) from bovine aorta. Next, we incubated [3H]inositol-labeled AR4-2J cells with cell permeant and non radiolabeled 2,5,6-tri-O-butyryl-myo-inositol 1,3, 4-trisphosphate hexakis(acetoxymethyl) ester. This treatment increased cellular levels of Ins(1,3,4)P3 2.7-fold, while [3H]Ins(3, 4,5,6)P4 levels increased 2-fold; there were no changes to levels of other 3H-labeled inositol phosphates. This experiment provides the first direct evidence that levels of Ins(3,4,5,6)P4 are regulated by Ins(1,3,4)P3 in vivo, independently of Ins(1,3,4)P3 being metabolized to Ins(3,4,5,6)P4. In addition, we found that the Ins(1, 3,4)P3 metabolites, namely Ins(1,3)P2 and Ins(3,4)P2, were >100-fold weaker inhibitors of the 1-kinase compared with Ins(1,3,4)P3 itself (IC50 = 0.17 microM). This result shows that dephosphorylation of Ins(1,3,4)P3 in vivo is an efficient mechanism to "switch-off" the cellular regulation of Ins(3,4,5,6)P4 levels that comes from Ins(1,3, 4)P3-mediated inhibition of the 1-kinase. We also found that Ins(1,3, 6)P3 and Ins(1,4,6)P3 were poor inhibitors of the 1-kinase (IC50 = 17 and >30 microM, respectively). The non-physiological trisphosphates, D/L Ins(1,2,4)P3, inhibited 1-kinase relatively potently (IC50 = 0.7 microM), thereby suggesting a new strategy for the rational design of therapeutically useful kinase inhibitors. Overall, our data provide new information to support the idea that Ins(1,3,4)P3 acts in an important signaling cascade. PMID- 10383397 TI - 3-deazaadenosine, a S-adenosylhomocysteine hydrolase inhibitor, has dual effects on NF-kappaB regulation. Inhibition of NF-kappaB transcriptional activity and promotion of IkappaBalpha degradation. AB - Previously we reported that 3-deazaadenosine (DZA), a potent inhibitor and substrate for S-adenosylhomocysteine hydrolase inhibits bacterial lipopolysaccharide-induced transcription of tumor necrosis factor-alpha and interleukin-1beta in mouse macrophage RAW 264.7 cells. In this study, we demonstrate the effects of DZA on nuclear factor-kappaB (NF-kappaB) regulation. DZA inhibits the transcriptional activity of NF-kappaB through the hindrance of p65 (Rel-A) phosphorylation without reduction of its nuclear translocation and DNA binding activity. The inhibitory effect of DZA on NF-kappaB transcriptional activity is potentiated by the addition of homocysteine. Taken together, DZA promotes the proteolytic degradation of IkappaBalpha, but not IkappaBbeta, resulting in an increase of DNA binding activity of NF-kappaB in the nucleus in the absence of its transcriptional activity in RAW 264.7 cells. The reduction of IkappaBalpha by DZA is neither involved in IkappaB kinase complex activation nor modulated by the addition of homocysteine. This study strongly suggests that DZA may be a potent drug for the treatment of diseases in which NF-kappaB plays a central pathogenic role, as well as a useful tool for studying the regulation and physiological functions of NF-kappaB. PMID- 10383398 TI - Mechanism of iron transport to the site of heme synthesis inside yeast mitochondria. AB - The import of metals, iron in particular, into mitochondria is poorly understood. Iron in mitochondria is required for the biosynthesis of heme and various iron sulfur proteins. We have developed an in vitro assay to follow the uptake of iron into isolated yeast mitochondria. By measuring the incorporation of iron into porphyrin by ferrochelatase in the matrix, we were able to define the mechanism of iron import. Iron uptake is driven energetically by a membrane potential across the inner membrane but does not require ATP. Only reduced iron is functional in generating heme. Iron cannot be preloaded in the mitochondrial matrix but rather has to be transported across the inner membrane simultaneously with the synthesis of heme, suggesting that ferrochelatase receives iron directly from the inner membrane. Transport of iron is inhibited by manganese but not by zinc, nickel, and copper ions, explaining why in vivo these ions are not incorporated into porphyrin. The inner membrane proteins Mmt1p and Mmt2p proposed to be involved in mitochondrial iron movement are not required for the supply of ferrochelatase with iron. Iron transport can be reconstituted efficiently in a membrane potential-dependent fashion in proteoliposomes that were formed from a detergent extract of mitochondria. Our biochemical analysis of iron import into yeast mitochondria provides the basis for the identification of components involved in transport. PMID- 10383399 TI - Differential pharmacological properties and signal transduction of the sphingosine 1-phosphate receptors EDG-1, EDG-3, and EDG-5. AB - Sphingosine 1-phosphate (SPP) is a potent lipid mediator released upon cellular activation. In this report, pharmacological properties of the three G-protein coupled receptors (GPCRs) for SPP, EDG-1, -3, and -5 are characterized using a Xenopus oocyte expression system, which lacks endogenous SPP receptors. Microinjection of the EDG-3 and EDG-5 but not EDG-1 mRNA conferred SPP-responsive intracellular calcium transients; however, the EDG-5 response was quantitatively much less. Co-expression of EDG-1 receptor with the chimeric Galphaqi protein conferred SPP responsiveness. Galphaqi or Galphaq co-injection also potentiated the EDG-5 and EDG-3 mediated responses to SPP. These data suggest that SPP receptors couple differentially to the Gq and Gi pathway. All three GPCRs were also activated by sphingosylphosphorylcholine, albeit at higher concentrations. None of the other related sphingolipids tested stimulated or blocked SPP-induced calcium responses. However, suramin, a polycyclic anionic compound, selectively antagonized SPP-activated calcium transients in EDG-3 expressing oocytes with an IC50 of 22 microM, suggesting that it is an antagonist selective for the EDG-3 GPCR isotype. We conclude that the three SPP receptors signal differentially by coupling to different G-proteins. Furthermore, because only EDG-3 was antagonized by suramin, variations in receptor structure may determine differences in antagonist selectivity. This property may be exploited to synthesize receptor subtype-specific antagonists. PMID- 10383400 TI - Direct interaction in T-cells between thetaPKC and the tyrosine kinase p59fyn. AB - The protein kinase C (PKC) family has been clearly implicated in T-cell activation as have several nonreceptor protein-tyrosine kinases associated with the T-cell receptor, including p59fyn. This report demonstrates that thetaPKC and p59fyn specifically interact in vitro, in the yeast two-hybrid system, and in T cells. Further indications of direct interaction are that p59fyn potentiates thetaPKC catalytic activity and that thetaPKC is a substrate for tyrosine phosphorylation by p59fyn. This interaction may account for the localization of thetaPKC following T-cell activation, pharmacological disruption of which results in specific cell-signaling defects. The demonstration of a physical interaction between a PKC and a protein-tyrosine kinase expands the class of PKC-anchoring proteins (receptors for activated C kinases (RACKs)) and demonstrates a direct connection between these two major T-cell-signaling pathways. PMID- 10383401 TI - Involvement of cell cycle elements, cyclin-dependent kinases, pRb, and E2F x DP, in B-amyloid-induced neuronal death. AB - Previous evidence by others has indicated that a variety of cell cycle-related molecules are up-regulated in brains of Alzheimer's disease patients. The significance of this increase, however, is unclear. Accordingly, we examined the obligate nature of cyclin-dependent kinases and select downstream targets of these kinases in death of neurons evoked by B-amyloid (AB) protein. We present pharmacological and molecular biological evidence that cyclin-dependent kinases, in particular Cdk4/6, are required for such neuronal death. In addition, we demonstrate that the substrate of Cdk4/6, pRb/p107, is phosphorylated during AB treatment and that one target of pRb/p107, the E2F x DP complex, is required for AB-evoked neuronal death. These results provide evidence that cell cycle elements play a required role in death of neurons evoked by AB and suggest that these elements play an integral role in Alzheimer's disease-related neuronal death. PMID- 10383402 TI - Yeast transcriptional regulator Leu3p. Self-masking, specificity of masking, and evidence for regulation by the intracellular level of Leu3p. AB - Recent work suggests that the masking of the activation domain (AD) of yeast transactivator Leu3p, observed in the absence of the metabolic signal alpha isopropylmalate, is an intramolecular event. Much of the evidence came from the construction and analysis of a mutant form of Leu3p (Leu3-dd) whose AD is permanently masked (Wang, D., Hu, Y., Zheng, F., Zhou, K., and Kohlhaw, G. B. (1997) J. Biol. Chem. 272, 19383-19392). In a modified two-hybrid experiment, the ADs of both wild type Leu3p and Leu3-dd were shown to interact with the remainder of the Leu3 protein, in an alpha-isopropylmalate-dependent manner. The finding that masking and unmasking proceed apparently normally when full-length Leu3p is expressed in mammalian cells is also consistent with the notion of intramolecular masking. Here we report on the identification of nine missense mutations (all of them suppressors of the Leu3-dd phenotype) that cause permanent unmasking of Leu3p. The nine mutations map to three short segments located within a 140 residue-long region of the C-terminal part of the middle region of Leu3p. These segments may be part of a spatial trap for the AD. We also performed "domain swaps" between Leu3p and Cha4p, a serine/threonine-responsive activator that, like Leu3p, belongs to the family of Zn(II)2Cys6 proteins. We show that AD masking and response to the appropriate metabolic signal only occur when a given AD remains attached to its own middle region; middle region swapping results in constitutively active proteins. Finally, we show that the extent to which Leu3p regulates reporter gene expression depends on the intracellular concentration of Leu3p. The possible physiological significance of this observation is discussed in light of the known regulation of Leu3p by Gcn4p. PMID- 10383403 TI - Association of atypical protein kinase C isotypes with the docker protein FRS2 in fibroblast growth factor signaling. AB - FRS2 is a docker protein that recruits signaling proteins to the plasma membrane in fibroblast growth factor signal transduction. We report here that FRS2 was associated with PKC lambda when Swiss 3T3 cells were stimulated with basic fibroblast growth factor. PKC zeta, the other member of the atypical PKC subfamily, could also bind FRS2. The association between FRS2 and PKC lambda is likely to be direct as shown by yeast two-hybrid analysis. The C-terminal fragments of FRS2 (amino acid residues 300-508) and SNT2 (amino acids 281-492), an isoform bearing 50% identity to FRS2, interacted with PKC lambda at a region (amino acids 240-562) that encompasses the catalytic domain. In vitro kinase assays revealed neither FRS2 nor SNT2 was a substrate of PKC lambda or zeta. Mutation of the alanine residue (Ala-120) to glutamate in the pseudo-substrate region of PKC lambda results in a constitutively active kinase that exhibited more than 2-fold greater binding to FRS2 in vitro than its "closed" wild-type counterpart. Tyrosine phosphorylation of FRS2 did not affect its binding to the constitutively active PKC lambda mutant, suggesting that the activation of PKC lambda is necessary and sufficient for its association with FRS2. It is likely that FRS2 serves as an anchoring protein for targeting activated atypical PKCs to the cell plasma membrane in signaling pathways. PMID- 10383404 TI - In vivo ligand specificity of E-selectin binding to multivalent sialyl Lewisx N linked oligosaccharides. AB - The in vivo specificity for E-selectin binding to a panel of N-linked oligosaccharides containing a clustered array of one to four sialyl Lewisx (SLex; NeuAcalpha2-3Gal[Fucalpha1-3]beta1-4GlcNAc) determinants was studied in mice. Following intraperitoneal dosing with lipopolysaccharide, radioiodinated tyrosinamide N-linked oligosaccharides were dosed i.v. and analyzed for their pharmacokinetics and biodistribution. Specific targeting was determined from the degree of SLex oligosaccharide targeting relative to a sialyl oligosaccharide control. Oligosaccharides targeted the kidney with the greatest selectivity after a 4-h induction period following lipopolysaccharide dosing. Unique pharmacokinetic profiles were identified for SLex biantennary and triantennary oligosaccharides but not for monovalent and tetraantennary SLex oligosaccharides or sialyl oligosaccharide controls. Biodistribution studies established that both SLex biantennary and triantennary oligosaccharides distributed to the kidney with 2-3-fold selectivity over sialyl oligosaccharide controls, whereas monovalent and tetraantennary SLex oligosaccharides failed to mediate specific kidney targeting. Simultaneous dosing of SLex biantennary or triantennary oligosaccharide with a mouse anti-E-selectin monoclonal antibody blocked kidney targeting, whereas co administration with anti-P-selectin monoclonal antibody did not significantly block kidney targeting. The results suggest that SLex biantennary and triantennary are N-linked oligosaccharide ligands for E-selectin and implicate E selectin as a bivalent receptor in the murine kidney endothelium. PMID- 10383405 TI - Circular permutation analysis as a method for distinction of functional elements in the M20 loop of Escherichia coli dihydrofolate reductase. AB - A functional element of an enzyme can be defined as the smallest unit of the local peptide backbone of which the connectivity is crucial for the catalytic activity. In order to elucidate the distribution of functional elements in an active site flexible loop (the M20 loop) of Escherichia coli dihydrofolate reductase, systematic cleavage of main chain connectivity was performed using circular permutation. Our analysis is based on the assumption that a permutation within a functional element would significantly reduce enzyme function, whereas ones outside or at the boundaries of the elements would affect the function only slightly. Thus, a functional element would be assigned as the minimum peptide chain between the identified boundaries. Comparison of the activities of the circularly permuted variants revealed that the peptide chain around the M20 loop could be divided into four regions (regions 1-4). Region 1 was found to play an important role in overall tertiary fold because most variants permuted at region 1 did not accumulate in E. coli cells stably. A distinction between region 2 and region 3 was in agreement with the extent of movements calculated from the coordinates of alpha carbons, supporting the idea that the movement of peptide backbone is a key feature of enzyme function. The boundary between region 3 and region 4 coincided with that between the M20 loop and the following alpha helix. From equilibrium binding studies, region 2 was found to be involved in the binding of nicotinamide substrates, whereas region 4 appeared to be very important for the binding of pterin substrates. PMID- 10383406 TI - Oxidizing side of the cyanobacterial photosystem I. Evidence for interaction between the electron donor proteins and a luminal surface helix of the PsaB subunit. AB - Photosystem I (PSI) interacts with plastocyanin or cytochrome c6 on the luminal side. To identify sites of interaction between plastocyanin/cytochrome c6 and the PSI core, site-directed mutations were generated in the luminal J loop of the PsaB protein from Synechocystis sp. PCC 6803. The eight mutant strains differed in their photoautotrophic growth. Western blotting with subunit-specific antibodies indicated that the mutations affected the PSI level in the thylakoid membranes. PSI proteins could not be detected in the S600R/G601C/N602I, N609K/S610C/T611I, and M614I/G615C/W616A mutant membranes. The other mutant strains contained different levels of PSI proteins. Among the mutant strains that contained PSI proteins, the H595C/L596I, Q627H/L628C/I629S, and N638C/N639S mutants showed similar levels of PSI-mediated electron transfer activity when either cytochrome c6 or an artificial electron donor was used. In contrast, cytochrome c6 could not function as an electron donor to the W622C/A623R mutant, even though the PSI activity mediated by an artificial electron donor was detected in this mutant. Thus, the W622C/A623R mutation affected the interaction of the PSI complex with cytochrome c6. Biotin-maleimide modification of the mutant PSI complexes indicated that His-595, Trp-622, Leu-628, Tyr-632, and Asn 638 in wild-type PsaB may be exposed on the surface of the PSI complex. The results presented here demonstrate the role of an extramembrane loop of a PSI core protein in the interaction with soluble electron donor proteins. PMID- 10383407 TI - A null mutation in murine CD36 reveals an important role in fatty acid and lipoprotein metabolism. AB - A null mutation in the scavenger receptor gene CD36 was created in mice by targeted homologous recombination. These mice produced no detectable CD36 protein, were viable, and bred normally. A significant decrease in binding and uptake of oxidized low density lipoprotein was observed in peritoneal macrophages of null mice as compared with those from control mice. CD36 null animals had a significant increase in fasting levels of cholesterol, nonesterified free fatty acids, and triacylglycerol. The increase in cholesterol was mainly within the high density lipoprotein fraction, while the increase in triacylglycerol was within the very low density lipoprotein fraction. Null animals had lower fasting serum glucose levels when compared with wild type controls. Uptake of 3H-labeled oleate was significantly reduced in adipocytes from null mice. However, the decrease was limited to the low ratios of fatty acid:bovine serum albumin, suggesting that CD36 was necessary for the high affinity component of the uptake process. The data provide evidence for a functional role for CD36 in lipoprotein/fatty acid metabolism that was previously underappreciated. PMID- 10383408 TI - Accumulation of sialic acid in endocytic compartments interferes with the formation of mature lysosomes. Impaired proteolytic processing of cathepsin B in fibroblasts of patients with lysosomal sialic acid storage disease. AB - The impact of an altered endocytic environment on the biogenesis of lysosomes was studied in fibroblasts of patients suffering from sialic acid storage disease (SASD). This inherited disorder is characterized by the accumulation of acidic monosaccharides in lysosomal compartments and a concomitant decrease of their buoyant density. We demonstrate that C-terminal trimming of the lysosomal cysteine proteinase cathepsin B is inhibited in SASD fibroblasts. This late event in the biosynthesis of cathepsin B normally takes place in mature lysosomes, suggesting an impaired biogenesis of these organelles in SASD cells. When normal fibroblasts are loaded with sucrose, which inhibits transport from late endosomes to lysosomes, C-terminal cathepsin B processing is prevented to the same extent. Further characterization of the terminal endocytic compartments of SASD cells revealed properties usually associated with late endosomes/prelysosomes. In addition to a decreased buoyant density, SASD "lysosomes" show a reduced acidification capacity and appear smaller than their normal counterparts. We conclude that the accumulation of small non-diffusible compounds within endocytic compartments interferes with the formation of mature lysosomes and that the acidic environment of the latter organelles is a prerequisite for C-terminal processing of lysosomal hydrolases. PMID- 10383409 TI - The membrane topology of proton-pumping Escherichia coli transhydrogenase determined by cysteine labeling. AB - The membrane topology of proton-pumping nicotinamide-nucleotide transhydrogenase from Escherichia coli was determined by site-specific chemical labeling. A His tagged cysteine-free transhydrogenase was used to introduce unique cysteines in positions corresponding to potential membrane loops. The cysteines were reacted with fluorescent reagents, fluorescein 5-maleimide or 2-[(4' maleimidyl)anilino]naphthalene-6-sulfonic acid, in both intact cells and inside out vesicles. Labeled transhydrogenase was purified with a small-scale procedure using a metal affinity resin, and the amount of labeling was measured as fluorescence on UV-illuminated acrylamide gels. The difference in labeling between intact cells and inside-out vesicles was used to discriminate between a periplasmic and a cytosolic location of the residues. The membrane region was found to be composed of 13 helices (four in the alpha-subunit and nine in the beta-subunit), with the C terminus of the alpha-subunit and the N terminus of the beta-subunit facing the cytosolic and periplasmic sides, respectively. These results differ from previous models with regard to both number of helices and the relative location and orientation of certain helices. This study constitutes the first in which all transmembrane segments of transhydrogenase have been experimentally determined and provides an explanation for the different topologies of the mitochondrial and E. coli transhydrogenases. PMID- 10383410 TI - The interaction between Z-DNA and the Zab domain of double-stranded RNA adenosine deaminase characterized using fusion nucleases. AB - Zab is a structurally defined protein domain that binds specifically to DNA in the Z conformation. It consists of amino acids 133-368 from the N terminus of human double-stranded RNA adenosine deaminase, which is implicated in RNA editing. Zab contains two motifs with related sequence, Zalpha and Zbeta. Zalpha alone is capable of binding Z-DNA with high affinity, whereas Zbeta alone has little DNA binding activity. Instead, Zbeta modulates Zalpha binding, resulting in increased sequence specificity for alternating (dCdG)n as compared with (dCdA/dTdG)n. This relative specificity has previously been demonstrated with short oligonucleotides. Here we demonstrate that Zab can also bind tightly to (dCdG)n stabilized in the Z form in supercoiled plasmids. Binding was assayed by monitoring cleavage of the plasmids using fusion nucleases, in which Z-DNA binding peptides from the N terminus of double-stranded RNA adenosine deaminase are linked to the nuclease domain of FokI. A fusion nuclease containing Zalpha shows less sequence specificity, as well as less conformation specificity, than one containing Zab. Further, a construct in which Zbeta has been replaced in Zab with Zalpha, cleaves Z-DNA regions in supercoiled plasmids more efficiently than the wild type but with little sequence specificity. We conclude that in the Zab domain, both Zalpha and Zbeta contact DNA. Zalpha contributes contacts that produce conformation specificity but not sequence specificity. In contrast, Zbeta contributes weakly to binding affinity but discriminates between sequences of Z DNAs. PMID- 10383411 TI - Mannose polyethylenimine conjugates for targeted DNA delivery into dendritic cells. AB - Cell surface-bound receptors represent suitable entry sites for gene delivery into cells by receptor-mediated endocytosis. Here we have taken advantage of the mannose receptor that is highly expressed on antigen-presenting dendritic cells for targeted gene transfer by employing mannosylpolyethylenimine (ManPEI) conjugates. Several ManPEI conjugates were synthesized and used for formation of ManPEI/DNA transfection complexes. Conjugates differed in the linker between mannose and polyethylenimine (PEI) and in the size of the PEI moiety. We demonstrate that ManPEI transfection is effective in delivering DNA into mannose receptor-expressing cells. Uptake of ManPEI/DNA complexes is receptor-specific, since DNA delivery can be competed with mannosylated albumin. Additionally, incorporation of adenovirus particles into transfection complexes effectively enhances transgene expression. This is particularly important for primary immunocompetent dendritic cells. It is demonstrated here that dendritic cells transfected with ManPEI/DNA complexes containing adenovirus particles are effective in activating T cells of T cell receptor transgenic mice in an antigen specific fashion. PMID- 10383412 TI - Brefeldin A increases the quantal size and alters the kinetics of catecholamine release from rat adrenal chromaffin cells. AB - The fungal metabolite, brefeldin A (BFA), is known to inhibit guanine nucleotide exchange on the ADP-ribosylating factors that are involved in vesicle membrane trafficking. Here, we investigated the action of BFA on Ca2+-regulated exocytosis in single rat adrenal chromaffin cells. Incubation of chromaffin cells with BFA (1 or 10 microM) for 2 h effectively disrupted the Golgi membranes but did not affect the pattern of catecholamine release triggered by high extracellular K+, which was monitored with carbon fiber amperometry along with cytosolic Ca2+ measurement. The BFA treatment, however, increased the mean quantal size of catecholamine-containing vesicles and the occurrence of amperometric events with a "foot" or "stand alone" signal (which reflects sluggish or incomplete dilation of the fusion pore). To examine whether BFA altered the Ca2+-dependence of exocytosis, we employed the whole-cell recording technique in conjunction with the capacitance measurement to measure exocytosis evoked from the entire cell during voltage-gated Ca2+ entry. Our results suggested that BFA treatment did not alter either the initial rate of capacitance increase or the total amount of capacitance increase. Therefore, in chromaffin cells, BFA treatment affects Ca2+ regulated exocytosis predominantly by increasing the quantal size and by slowing the fusion kinetics of some vesicles. PMID- 10383413 TI - Functional interaction between Oct-1 and retinoid X receptor. AB - The retinoid X receptor (RXR) is a member of the nuclear hormone receptor superfamily and heterodimerizes with a variety of other family members such as the thyroid hormone receptor (TR),1 retinoic acid receptor, vitamin D receptor, and peroxisome proliferator-activated receptor. Therefore, RXR is supposed to play a key role in a ligand-dependent regulation of gene transcription by nuclear receptors. In this study, we have identified the octamer-binding transcription factor-1 (Oct-1) as a novel interaction factor of RXR. In vitro pull-down assays using RXR deletion mutants showed that the interaction surfaces were located in the region encompassing the DNA binding domain (C domain) and the hinge domain (D domain) of RXR. We also showed that RXR interacted with the POU homeodomain but not with the POU-specific domain of Oct-1. Gel shift analysis revealed that Oct-1 reduced the binding of TR/RXR heterodimers to the thyroid hormone response element (TRE). In transient transfection assays using COS1 cells, Oct-1 repressed the T3-dependent transcriptional activity of TR/RXR heterodimers, consistent with in vitro DNA binding data; however, transcriptional activation by Gal4-TR(LBD) (LBD, ligand binding domain), which lacks its own DNA binding domain but retains responsiveness to T3, was not influenced by Oct-1. These results suggest that Oct 1 functionally interacts with RXR and negatively regulates the nuclear receptor signaling pathway by altering the DNA binding ability of the receptors. PMID- 10383414 TI - Functional characterization of the D-Tyr-tRNATyr deacylase from Escherichia coli. AB - The yihZ gene of Escherichia coli is shown to produce a deacylase activity capable of recycling misaminoacylated D-Tyr-tRNATyr. The reaction is specific and, under optimal in vitro conditions, proceeds at a rate of 6 s-1 with a Km value for the substrate equal to 1 microM. Cell growth is sensitive to interruption of the yihZ gene if D-tyrosine is added to minimal culture medium. Toxicity of exogenous D-tyrosine is exacerbated if, in addition to the disruption of yihZ, the gene of D-amino acid dehydrogenase (dadA) is also inactivated. Orthologs of the yihZ gene occur in many, but not all, bacteria. In support of the idea of a general role of the D-Tyr-tRNATyr deacylase function in the detoxification of cells, similar genes can be recognized in Saccharomyces cerevisiae, Caenorhabditis elegans, Arabidopsis thaliana, mouse, and man. PMID- 10383415 TI - Protein kinase C delta is essential for etoposide-induced apoptosis in salivary gland acinar cells. AB - We have previously shown that parotid C5 salivary acinar cells undergo apoptosis in response to etoposide treatment as indicated by alterations in cell morphology, caspase-3 activation, DNA fragmentation, sustained activation of c Jun N-terminal kinase, and inactivation of extracellular regulated kinases 1 and 2. Here we report that apoptosis results in the caspase-dependent cleavage of protein kinase C-delta (PKCdelta) to a 40-kDa fragment, the appearance of which correlates with a 9-fold increase in PKCdelta activity. To understand the function of activated PKCdelta in apoptosis, we have used the PKCdelta-specific inhibitor, rottlerin. Pretreatment of parotid C5 cells with rottlerin prior to the addition of etoposide blocks the appearance of the apoptotic morphology, the sustained activation of c-Jun N-terminal kinase, and inactivation of extracellular regulated kinases 1 and 2. Inhibition of PKCdelta also partially inhibits caspase-3 activation and DNA fragmentation. Immunoblot analysis shows that the PKCdelta cleavage product does not accumulate in parotid C5 cells treated with rottlerin and etoposide together, suggesting that the catalytic activity of PKCdelta may be required for cleavage. PKCalpha and PKCbeta1 activities also increase during etoposide-induced apoptosis. Inhibition of these two isoforms with Go6976 slightly suppresses the apoptotic morphology, caspase-3 activation, and DNA fragmentation, but has no effect on the sustained activation of c-Jun N-terminal kinase or inactivation of extracellular regulated kinase 1 and 2. These data demonstrate that activation of PKCdelta is an integral and essential part of the apoptotic program in parotid C5 cells and that specific activated isoforms of PKC may have distinct functions in cell death. PMID- 10383416 TI - Effect of the epsilon-subunit on nucleotide binding to Escherichia coli F1-ATPase catalytic sites. AB - The influence of the epsilon-subunit on the nucleotide binding affinities of the three catalytic sites of Escherichia coli F1-ATPase was investigated, using a genetically engineered Trp probe in the adenine-binding subdomain (beta-Trp-331). The interaction between epsilon and F1 was not affected by the mutation. Kd for binding of epsilon to betaY331W mutant F1 was approximately 1 nM, and epsilon inhibited ATPase activity by 90%. The only nucleotide binding affinities that showed significant differences in the epsilon-depleted and epsilon-replete forms of the enzyme were those for MgATP and MgADP at the high-affinity catalytic site 1. Kd1(MgATP) and Kd1(MgADP) were an order of magnitude higher in the absence of epsilon than in its presence. In contrast, the binding affinities for MgATP and MgADP at sites 2 and 3 were similar in the epsilon-depleted and epsilon-replete enzymes, as were the affinities at all three sites for free ATP and ADP. Comparison of MgATP binding and hydrolysis parameters showed that in the presence as well as the absence of epsilon, Km equals Kd3. Thus, in both cases, all three catalytic binding sites have to be occupied to obtain rapid (Vmax) MgATP hydrolysis rates. PMID- 10383417 TI - Possible involvement of a novel STAM-associated molecule "AMSH" in intracellular signal transduction mediated by cytokines. AB - STAM containing an SH3 (Src homology 3) domain and an immunoreceptor tyrosine based activation motif was previously revealed to be implicated in signaling pathways immediately downstream of Jak2 and Jak3 tyrosine kinases associated with cytokine receptors. We molecularly cloned a novel molecule interacting with the SH3 domain of STAM, which was named AMSH (associated molecule with the SH3 domain of STAM). AMSH contains a putative bipartite nuclear localization signal and a homologous region of a c-Jun activation domain-binding protein 1 (JAB1) subdomain in addition to a binding site for the SH3 domain of STAM. AMSH mutant deleted of the C-terminal half conferred dominant negative effects on signaling for DNA synthesis and c-myc induction mediated by interleukin 2 and granulocyte macrophage-colony-stimulating factor. These results suggest that AMSH plays a critical role in the cytokine-mediated intracellular signal transduction downstream of the Jak2/Jak3.STAM complex. PMID- 10383418 TI - Characterization of DP103, a novel DEAD box protein that binds to the Epstein Barr virus nuclear proteins EBNA2 and EBNA3C. AB - The Epstein-Barr virus-encoded nuclear antigens EBNA2 and EBNA3C both interact with the cellular transcription factor RBP-Jkappa and modulate the expression of several shared target genes, suggesting a tight cooperation in latently infected cells. In a survey for additional cellular factors that bind to EBNA2 as well as EBNA3C, we have isolated and characterized DP103, a novel human member of the DEAD box family of putative ATP-dependent RNA helicases. The interaction with DP103 is mediated by amino acids (aa) 121-213 of EBNA2 and aa 534-778 of EBNA3C, regions that are not involved in binding of the viral proteins to RBP-Jkappa. The DP103-cDNA encodes a protein of 824 aa that harbors all of the common DEAD box motifs. Monoclonal antibodies raised against DP103 detect a protein of 103 kDa in mammalian cells that resides in high molecular weight complexes in vivo. We have detected an ATPase activity intrinsic to or closely associated with DP103. By subcellular fractionation, we find DP103 in both a soluble nuclear fraction as well as in the insoluble skeletal fraction. Whereas the protein and its mRNA are uniformly expressed in all tested cell lines, we observed differential expression of the mRNA in normal human tissues. PMID- 10383419 TI - Keratin filament suspensions show unique micromechanical properties. AB - All epithelial cells feature a prominent keratin intermediate filament (IF) network in their cytoplasm. Studies in transgenic mice and in patients with inherited epithelial fragility syndromes showed that a major function of keratin IFs is to provide mechanical support to epithelial cell sheets. Yet the micromechanical properties of keratin IFs themselves remain unknown. We used rheological methods to assess the properties of suspensions of epidermal type I and type II keratin IFs and of vimentin, a type III IF polymer. We find that both types of IFs form gels with properties akin to visco-elastic solids. With increasing deformation they display strain hardening and yield relatively rapidly. Remarkably, both types of gels recover their preshear properties upon cessation of the deformation. Repeated imposition of small deformations gives rise to a progressively stiffer gel for keratin but not vimentin IFs. The visco elastic moduli of both gels show a weak dependence upon the frequency of the input shear stress and the concentration of the polymer, suggesting that both steric and nonsteric interactions between individual polymers contribute to the observed mechanical properties. In support of this, the length of individual polymers contributes only modestly to the properties of IF gels. Collectively these properties render IFs unique among cytoskeletal polymers and have strong implications for their function in vivo. PMID- 10383420 TI - Cloning and recombinant expression of a novel mouse-secreted phospholipase A2. AB - Secreted phospholipases A2 (sPLA2s) form a class of structurally related enzymes that are involved in a variety of physiological and pathological effects including inflammation and associated diseases, cell proliferation, cell adhesion, and cancer, and are now known to bind to specific membrane receptors. Here, we report the cloning and expression of a novel sPLA2 isolated from mouse thymus. Based on its structural features, this sPLA2 is most similar to the previously cloned mouse group IIA sPLA2 (mGIIA sPLA2). As for mGIIA sPLA2, the novel sPLA2 is made up of 125 amino acids with 14 cysteines, is basic (pI = 8.71) and its gene has been mapped to mouse chromosome 4. However, the novel sPLA2 has only 48% identity with mGIIA and displays similar levels of identity with the other mouse group IIC and V sPLA2s, indicating that the novel sPLA2 is not an isoform of mGIIA sPLA2. This novel sPLA2 has thus been called mouse group IID (mGIID) sPLA2. In further contrast with mGIIA, which is found mainly in intestine, transcripts coding for mGIID sPLA2 are found in several tissues including pancreas, spleen, thymus, skin, lung, and ovary, suggesting distinct functions for the two enzymes. Recombinant expression of mGIID sPLA2 in Escherichia coli indicates that the cloned sPLA2 is an active enzyme that has much lower specific activity than mGIIA and displays a distinct specificity for binding to various phospholipid vesicles. Finally, recombinant mGIID sPLA2 did not bind to the mouse M-type sPLA2 receptor, while mGIIA was previously found to bind to this receptor with high affinity. PMID- 10383421 TI - Identification of an interaction between residue 6 of the natural peptide ligand and a distinct residue within the amino-terminal tail of the secretin receptor. AB - Photoaffinity labeling is a powerful tool for the characterization of the molecular basis of ligand binding. We recently used this technique to demonstrate the proximity between a residue within the carboxyl-terminal half of a secretin like ligand and the amino-terminal domain of the secretin receptor (Dong, M., Wang, Y., Pinon, D. I., Hadac, E. M., and Miller, L. J. (1999) J. Biol. Chem. 274, 903-909). In this work, we have developed another novel radioiodinatable secretin analogue ([Bpa6,Tyr10]rat secretin-27) that incorporates a photolabile p benzoyl-L-phenylalanine (Bpa) residue into position 6 of the amino-terminal half of the ligand and used this to identify a specific receptor residue proximate to it. This probe specifically bound to the secretin receptor with high affinity (IC50 = 13.2 +/- 2.5 nM) and was a potent stimulant of cAMP accumulation in secretin receptor-bearing Chinese hamster ovary-SecR cells (EC50 = 720 +/- 230 pM). It covalently labeled the secretin receptor in a saturable and specific manner. Cyanogen bromide cleavage of this molecule yielded a single labeled fragment that migrated on an SDS-polyacrylamide gel at Mr = 19,000 that shifted to 10 after deglycosylation, most consistent with either of two glycosylated fragments within the amino-terminal tail. By immunoprecipitation with antibody directed to epitope tags incorporated into each of the two candidate fragments, the most distal fragment at the amino terminus was identified as the domain of labeling. The labeled domain was further refined to the first 16 residues by endoproteinase Lys-C cleavage and by cyanogen bromide cleavage of another receptor construct in which Val16 was mutated to Met. Radiochemical sequencing of photoaffinity-labeled secretin receptor fragments established that Val4 was the specific site of covalent attachment. This provides the first residue-residue contact between a secretin ligand and its receptor and will contribute substantially to the molecular understanding of this interaction. PMID- 10383422 TI - Surfactant protein B (SP-B) -/- mice are rescued by restoration of SP-B expression in alveolar type II cells but not Clara cells. AB - Surfactant protein B (SP-B) mRNA and protein are restricted to alveolar Type II and Clara cells in the respiratory epithelium. In order to investigate the function of SP-B in these distinct cell types, transgenic mice were generated in which SP-B expression was selectively restored in Type II cells or Clara cells of SP-B -/- mice. The 4.8-kilobase murine SP-C promoter was used to generate 3 transgenic lines which expressed human SP-B in Type II cells (mSP-C/hSP-B). Likewise, the 2.3-kilobase murine CCSP promoter was used to generate two transgenic lines which expressed human SP-B in Clara cells (mCCSP/hSP-B). mSP C/hSP-B and mCCSP/hSP-B transgenic mice were subsequently bred to SP-B +/- mice in order to selectively express SP-B in Type II cells or Clara cells of SP-B -/- mice. Selective restoration of SP-B expression in Type II cells completely rescued the neonatal lethal phenotype in SP-B -/- mice. Expression of SP-B in some, but not all Type II cells of SP-B -/- mice, allowed postnatal survival, but resulted in significantly altered lung architecture and function. Selective restoration of SP-B expression in Clara cells of SP-B -/- mice resulted in respiratory dysfunction and invariable neonatal death, related to the complete absence of mature SP-B peptide in these mice. These results indicate that expression and processing of the SP-B proprotein to the mature peptide in Type II cells is absolutely required for lung function in vivo and that SP-B expression in Clara cells cannot substitute for this function. PMID- 10383423 TI - Mutations in the active site of penicillin-binding protein PBP2x from Streptococcus pneumoniae. Role in the specificity for beta-lactam antibiotics. AB - Penicillin-binding protein 2x (PBP2x) isolated from clinical beta-lactam resistant strains of Streptococcus pneumoniae (R-PBP2x) have a reduced affinity for beta-lactam antibiotics. Their transpeptidase domain carries numerous substitutions compared with homologous sequences from beta-lactam-sensitive streptococci (S-PBP2x). Comparison of R-PBP2x sequences suggested that the mutation Gln552 --> Glu is important for resistance development. Mutants selected in the laboratory with cephalosporins frequently contain a mutation Thr550 --> Ala. The high resolution structure of a complex between S-PBP2x* and cefuroxime revealed that Gln552 and Thr550, which belong to strand beta3, are in direct contact with the cephalosporin. We have studied the effect of alterations at positions 552 and 550 in soluble S-PBP2x (S-PBP2x*) expressed in Escherichia coli. Mutation Q552E lowered the acylation efficiency for both penicillin G and cefotaxime when compared with S-PBP2x*. We propose that the introduction of a negative charge in strand beta3 conflicts with the negative charge of the beta lactam. Mutation T550A lowered the acylation efficiency of the protein for cefotaxime but not for penicillin G. The in vitro data presented here are in agreement with the distinct resistance profiles mediated by these mutations in vivo and underline their role as powerful resistance determinants. PMID- 10383424 TI - Lys13 plays a crucial role in the functional adaptation of the thermophilic triose-phosphate isomerase from Bacillus stearothermophilus to high temperatures. AB - The thermophilic triose-phosphate isomerases (TIMs) of Bacillus stearothermophilus (bTIM) and Thermotoga maritima (tTIM) have been found to possess a His12-Lys13 pair instead of the Asn12-Gly13 pair normally present in mesophilic TIMs. His12 in bTIM was proposed to prevent deamidation at high temperature, while the precise role of Lys13 is unknown. To investigate the role of the His12 and Lys13 pair in the enzyme's thermoadaptation, we reintroduced the "mesophilic residues" Asn and Gly into both thermophilic TIMs. Neither double mutant displayed diminished structural stability, but the bTIM double mutant showed drastically reduced catalytic activity. No similar behavior was observed with the tTIM double mutant, suggesting that the presence of the His12 and Lys13 cannot be systematically correlated to thermoadaptation in TIMs. We determined the crystal structure of the bTIM double mutant complexed with 2-phosphoglycolate to 2.4-A resolution. A molecular dynamics simulation showed that upon substitution of Lys13 to Gly an increase of the flexibility of loop 1 is observed, causing an incorrect orientation of the catalytic Lys10. This suggests that Lys13 in bTIM plays a crucial role in the functional adaptation of this enzyme to high temperature. Analysis of bTIM single mutants supports this assumption. PMID- 10383425 TI - Molecular cloning and characterization of MT-ACT48, a novel mitochondrial acyl CoA thioesterase. AB - While characterizing Eps15 partners, we identified a 48-kDa polypeptide (p48) which was precipitated by Eps15-derived glutathione S-transferase fusion proteins. A search in a murine expressed sequence tag data base with N-terminal microsequences of p48 led to the identification of two complete cDNA clones encoding two isoforms of a 439-amino acid protein sharing 95% nucleic and amino acid identity. Northern blot and immunoblotting studies showed that p48 was ubiquitously expressed. A significant homology (19% identity and 40% similarity) between p48 and rat brain cytosolic acyl-CoA thioesterase was observed in an 80 amino acid C-terminal domain, retrieved from proteins from human, nematode, and plants. The thioesterase function of p48 was further demonstrated against long chain acyl-CoAs in a spectrophotometric assay. Furthermore, data obtained from sequence analysis showed that p48 contained a mitochondrial targeting signal, cleaved in mature protein as assessed by microsequencing. The mitochondrial localization of both endogenous and transfected p48 was confirmed by confocal microscopy. These results indicate that p48, called MT-ACT48 (mitochondrial acyl CoA thioesterase of 48 kDa), defines a novel family of mitochondrial long chain acyl-CoA thioesterases. PMID- 10383426 TI - Inhibition of mammalian legumain by some cystatins is due to a novel second reactive site. AB - We have investigated the inhibition of the recently identified family C13 cysteine peptidase, pig legumain, by human cystatin C. The cystatin was seen to inhibit enzyme activity by stoichiometric 1:1 binding in competition with substrate. The Ki value for the interaction was 0.20 nM, i.e. cystatin C had an affinity for legumain similar to that for the papain-like family C1 cysteine peptidase, cathepsin B. However, cystatin C variants with alterations in the N terminal region and the "second hairpin loop" that rendered the cystatin inactive against cathepsin B, still inhibited legumain with Ki values 0.2-0.3 nM. Complexes between cystatin C and papain inhibited legumain activity against benzoyl-Asn-NHPhNO2 as efficiently as did cystatin C alone. Conversely, cystatin C inhibited papain activity against benzoyl-Arg-NHPhNO2 whether or not the cystatin had been incubated with legumain, strongly indicating that the cystatin inhibited the two enzymes with non-overlapping sites. A ternary complex between legumain, cystatin C, and papain was demonstrated by gel filtration supported by immunoblotting. Screening of a panel of cystatin superfamily members showed that type 1 inhibitors (cystatins A and B) and low Mr kininogen (type 3) did not inhibit pig legumain. Of human type 2 cystatins, cystatin D was non-inhibitory, whereas cystatin E/M and cystatin F displayed strong (Ki 0.0016 nM) and relatively weak (Ki 10 nM) affinity for legumain, respectively. Sequence alignments and molecular modeling led to the suggestion that a loop located on the opposite side to the papain-binding surface, between the alpha-helix and the first strand of the main beta-pleated sheet of the cystatin structure, could be involved in legumain binding. This was corroborated by analysis of a cystatin C variant with substitution of the Asn39 residue in this loop (N39K-cystatin C); this variant showed a slight reduction in affinity for cathepsin B (Ki 1.5 nM) but >>5,000-fold lower affinity for legumain (Ki >>1,000 nM) than wild-type cystatin C. PMID- 10383427 TI - A direct role for the macrophage low density lipoprotein receptor in atherosclerotic lesion formation. AB - To evaluate the contribution of the macrophage low density lipoprotein receptor (LDLR) to atherosclerotic lesion formation, we performed bone marrow transplantation studies in different mouse strains. First, LDLR(-/-) mice were transplanted with either LDLR(+/+) marrow or LDLR(-/-) marrow and were challenged with an atherogenic Western type diet. The diet caused severe hypercholesterolemia of a similar degree in the two groups, and no differences in the aortic lesion area were detected. Thus, macrophage LDLR expression does not influence foam cell lesion formation in the setting of extreme LDL accumulation. To determine whether macrophage LDLR expression affects foam cell formation under conditions of moderate, non-LDL hyperlipidemia, we transplanted C57BL/6 mice with either LDLR(-/-) marrow (experimental group) or LDLR(+/+) marrow (controls). Cholesterol levels were not significantly different between the two groups at baseline or after 6 weeks on a butterfat diet, but were 40% higher in the experimental mice after 13 weeks, mostly due to accumulation of beta-very low density lipoprotein (beta-VLDL). Despite the increase in cholesterol levels, mice receiving LDLR(-/-) marrow developed 63% smaller lesions than controls, demonstrating that macrophage LDLR affects the rate of foam cell formation when the atherogenic stimulus is beta-VLDL. We conclude that the macrophage LDLR is responsible for a significant portion of lipid accumulation in foam cells under conditions of dietary stress. PMID- 10383428 TI - Activation of the c-Jun N-terminal kinase/stress-activated protein kinase pathway by overexpression of caspase-8 and its homologs. AB - Caspase-8 is the most proximal caspase in the caspase cascade and possesses a prodomain consisting of two homologous death effector domains (DEDs). We have discovered that caspase-8 and its homologs can physically interact with tumor necrosis factor receptor-associated factor family members and activate the c-Jun N-terminal kinase (JNK, or stress-activated protein kinase) pathway. This ability resides in the DED-containing prodomain of these proteins and is independent of their role as cell death proteases. A point mutant in the first DED of caspase-8 can block JNK activation induced by several death domain receptors. Inhibition of JNK activation blocks apoptosis mediated by caspase-10, Mach-related inducer of toxicity/cFLIP, and Fas/CD95, thereby suggesting a cooperative role of this pathway in the mediation of caspase-induced apoptosis. PMID- 10383429 TI - Disassembly of the cytosolic chaperonin in mammalian cell extracts at intracellular levels of K+ and ATP. AB - The eukaryotic, cytoplasmic chaperonin, CCT, is essential for the biogenesis of actin- and tubulin-based cytoskeletal structures. CCT purifies as a doubly toroidal particle containing two eight-membered rings of approximately 60-kDa ATPase subunits, each encoded by an essential and highly conserved gene. However, immunofluorescence detection with subunit-specific antibodies has indicated that in cells CCT subunits do not always co-localize. We report here that CCT ATPase activity is highly dependent on K+ ion concentration and that in cell extracts, at physiological levels of K+ and ATP, there is considerable dissociation of CCT to a smaller oligomeric structure and free subunits. This dissociation is consequent to ATP hydrolysis and is readily reversed on removal of ATP. The ranking order for ease with which subunits can exit the chaperonin particle correlates well with the length of a loop structure, identified by homology modeling, in the intermediate domain of CCT subunits. K+-ATP-induced disassembly is not an intrinsic property of purified CCT over a 40-fold concentration range and requires the presence of additional factor(s) present in cell extracts. PMID- 10383430 TI - Peroxisomal and mitochondrial fatty acid beta-oxidation in mice nullizygous for both peroxisome proliferator-activated receptor alpha and peroxisomal fatty acyl CoA oxidase. Genotype correlation with fatty liver phenotype. AB - Fatty acid beta-oxidation occurs in both mitochondria and peroxisomes. Long chain fatty acids are also metabolized by the cytochrome P450 CYP4A omega-oxidation enzymes to toxic dicarboxylic acids (DCAs) that serve as substrates for peroxisomal beta-oxidation. Synthetic peroxisome proliferators interact with peroxisome proliferator activated receptor alpha (PPARalpha) to transcriptionally activate genes that participate in peroxisomal, microsomal, and mitochondrial fatty acid oxidation. Mice lacking PPARalpha (PPARalpha-/-) fail to respond to the inductive effects of peroxisome proliferators, whereas those lacking fatty acyl-CoA oxidase (AOX-/-), the first enzyme of the peroxisomal beta-oxidation system, exhibit extensive microvesicular steatohepatitis, leading to hepatocellular regeneration and massive peroxisome proliferation, implying sustained activation of PPARalpha by natural ligands. We now report that mice nullizygous for both PPARalpha and AOX (PPARalpha-/- AOX-/-) failed to exhibit spontaneous peroxisome proliferation and induction of PPARalpha-regulated genes by biological ligands unmetabolized in the absence of AOX. In AOX-/- mice, the hyperactivity of PPARalpha enhances the severity of steatosis by inducing CYP4A family proteins that generate DCAs and since they are not metabolized in the absence of peroxisomal beta-oxidation, they damage mitochondria leading to steatosis. Blunting of microvesicular steatosis, which is restricted to few liver cells in periportal regions in PPARalpha-/- AOX-/- mice, suggests a role for PPARalpha-induced genes, especially members of CYP4A family, in determining the severity of steatosis in livers with defective peroxisomal beta-oxidation. In age matched PPARalpha-/- mice, a decrease in constitutive mitochondrial beta oxidation with intact constitutive peroxisomal beta-oxidation system contributes to large droplet fatty change that is restricted to centrilobular hepatocytes. These data define a critical role for both PPARalpha and AOX in hepatic lipid metabolism and in the pathogenesis of specific fatty liver phenotype. PMID- 10383431 TI - A cluster of positively charged amino acids in the C4BP alpha-chain is crucial for C4b binding and factor I cofactor function. AB - C4b-binding protein (C4BP) is a regulator of the classical complement pathway, acting as a cofactor to factor I in the degradation of C4b. Computer modeling and structural analysis predicted a cluster of positively charged amino acids at the interface between complement control protein modules 1 and 2 of the C4BP alpha chain to be involved in C4b binding. Three C4BP mutants, R39Q, R64Q/R66Q, and R39Q/R64Q/R66Q, were expressed and assayed for their ability to bind C4b and to function as factor I cofactors. The apparent affinities of R39Q, R64Q/R66Q, and R39Q/R64Q/R66Q for immobilized C4b were 15-, 50-, and 140-fold lower, respectively, than that of recombinant wild type C4BP. The C4b binding site demonstrated herein was also found to be a specific heparin binding site. In C4b degradation, the mutants demonstrated decreased ability to serve as factor I cofactors. In particular, the R39Q/R64Q/R66Q mutant was inefficient as cofactor for cleavage of the Arg937-Thr938 peptide bond in C4b. In contrast, the factor I mediated cleavage of Arg1317-Asn1318 bond was less affected by the C4BP mutations. In conclusion, we identify a cluster of amino acids that is part of a C4b binding site involved in the regulation of the complement system. PMID- 10383432 TI - Transient effect of platelet-derived growth factor on GLUT4 translocation in 3T3 L1 adipocytes. AB - We earlier developed a novel method to detect translocation of the glucose transporter (GLUT) directly and simply using c-MYC epitope-tagged GLUT (GLUTMYC). To define the effect of platelet-derived growth factor (PDGF) on glucose transport in 3T3-L1 adipocytes, we investigated the PDGF- and insulin-induced glucose uptake, translocation of glucose transporters, and phosphatidylinositol (PI) 3-kinase activity in 3T3-L1, 3T3-L1GLUT4MYC, and 3T3-L1GLUT1MYC adipocytes. Insulin and PDGF stimulated glucose uptake by 9-10- and 5.5-6.5-fold, respectively, in both 3T3-L1 and 3T3-L1GLUT4MYC adipocytes. Exogenous GLUT4MYC expression led to enhanced PDGF-induced glucose transport. In 3T3-L1GLUT4MYC adipocytes, insulin and PDGF induced an 8- and 5-fold increase in GLUT4MYC translocation, respectively, determined in a cell-surface anti-c-MYC antibody binding assay. This PDGF-induced GLUT4MYC translocation was further demonstrated with fluorescent detection. In contrast, PDGF stimulated a 2-fold increase of GLUT1MYC translocation and 2.5-fold increase of glucose uptake in 3T3-L1GLUT1MYC adipocytes. The PDGF-induced GLUT4MYC translocation, glucose uptake, and PI 3 kinase activity were maximal (100%) at 5-10 min and thereafter rapidly declined to 40, 30, and 12%, respectively, within 60 min, a time when effects of insulin were maximal. Wortmannin (0.1 microM) abolished PDGF-induced GLUT4MYC translocation and glucose uptake in 3T3-L1GLUT4MYC adipocytes. These results suggest that PDGF can transiently trigger the translocation of GLUT4 and stimulate glucose uptake by translocation of both GLUT4 and GLUT1 in a PI 3 kinase-dependent signaling pathway in 3T3-L1 adipocytes. PMID- 10383433 TI - Regulation of lens fiber cell differentiation by transcription factor c-Maf. AB - To elucidate the regulatory mechanisms underlying lens development, we searched for members of the large Maf family, which are expressed in the mouse lens, and found three, c-Maf, MafB, and Nrl. Of these, the earliest factor expressed in the lens was c-Maf. The expression of c-Maf was most prominent in lens fiber cells and persisted throughout lens development. To examine the functional contribution of c-Maf to lens development, we isolated genomic clones encompassing the murine c-maf gene and carried out its targeted disruption. Insertion of the beta galactosidase (lacZ) gene into the c-maf locus allowed visualization of c-Maf accumulation in heterozygous mutant mice by staining for LacZ activity. Homozygous mutant embryos and newborns lacked normal lenses. Histological examination of these mice revealed defective differentiation of lens fiber cells. The expression of crystallin genes was severely impaired in the c-maf-null mutant mouse lens. These results demonstrate that c-Maf is an indispensable regulator of lens differentiation during murine development. PMID- 10383434 TI - Novel low molecular weight microtubule-associated protein-2 isoforms contain a functional nuclear localization sequence. AB - Known high and low molecular weight (LMW) MAP2 protein isoforms result from alternative splicing of the MAP2 gene. Contrary to previous reports that MAP2 is neural-specific, we recently identified MAP2 mRNA and protein in somatic and germ cells of rat testis, and showed the predominant testicular isoform is LMW. Although cytoplasmic in neural tissue, MAP2 appeared predominantly nuclear in germ cells using immunohistochemistry. We sought to determine whether this unexpected localization was due to the inclusion of exon 10 within novel LMW MAP2 isoforms. Normally excluded from the LMW MAP2c, exon 10 harbors a putative CcN motif, comprising a nuclear localization sequence (NLS) flanked by regulatory phosphorylation sites for protein kinase CK2 and cdc2 kinase. Characterization of MAP2 mRNA in adult and immature brain and testis, by reverse transcriptase polymerase chain reaction/Southern analysis and Northern blot, identified novel LMW forms containing exons 10 and 11, previously detected only in high molecular weight MAP2a and 2b. The MAP2 NLS targeted a large heterologous protein to the nucleus, as demonstrated using bacterially expressed MAP2-CcN-beta-galactosidase fusion protein and an in vitro nuclear import assay. Antibodies raised against the fusion protein produced a testicular immunohistochemical staining pattern correlating with MAP2 protein distribution in the nucleus of most germ cells, and precipitated both approximately 70-kDa and >220-kDa proteins recognized by the commercial MAP2-specific HM2 monoclonal antibody, supporting our hypothesis of a novel LMW MAP2 isoform. These results demonstrate the presence of a functional NLS in MAP2 and indicate that novel LMW MAP2 isoforms may be targeted to the nucleus in both neural and non-neuronal tissues. PMID- 10383435 TI - DNA bending by EcoRI DNA methyltransferase accelerates base flipping but compromises specificity. AB - EcoRI DNA methyltransferase was previously shown to bend its cognate DNA sequence by 52 degrees and stabilize the target adenine in an extrahelical orientation. We describe the characterization of an EcoRI DNA methyltransferase mutant in which histidine 235 was selectively replaced with asparagine. Steady-state kinetic and thermodynamic parameters for the H235N mutant revealed only minor functional consequences: DNA binding affinity (KDDNA) was reduced 10-fold, and kcat was decreased 30%. However, in direct contrast to the wild type enzyme, DNA bending within the mutant enzyme-DNA complexes was not observed by scanning force microscopy. The bending-deficient mutant showed enhanced discrimination against the methylation at nontarget sequence DNA. This enhancement of enzyme discrimination was accompanied by a change in the rate-limiting catalytic step. No presteady-state burst of product formation was observed, indicating that the chemistry step (or prior event) had become rate-limiting for methylation. Direct observation of the base flipping transition showed that the lack of burst kinetics was entirely due to slower base flipping. The combined data show that DNA bending contributes to the correct assembly of the enzyme-DNA complex to accelerate base flipping and that slowing the rate of this precatalytic isomerization can enhance specificity. PMID- 10383436 TI - Temperature adaptation of glutathione S-transferase P1-1. A case for homotropic regulation of substrate binding. AB - Human glutathione S-transferase P1-1 (GST P1-1) is a homodimeric enzyme expressed in several organs as well as in the upper layers of epidermis, playing a role against carcinogenic and toxic compounds. A sophisticated mechanism of temperature adaptation has been developed by this enzyme. In fact, above 35 degrees C, glutathione (GSH) binding to GST P1-1 displays positive cooperativity, whereas negative cooperativity occurs below 25 degrees C. This binding mechanism minimizes changes of GSH affinity for GST P1-1 because of temperature fluctuation. This is a likely advantage for epithelial skin cells, which are naturally exposed to temperature variation and, incidentally, to carcinogenic compounds, always needing efficient detoxifying systems. As a whole, GST P1-1 represents the first enzyme which displays a temperature-dependent homotropic regulation of substrate (e.g. GSH) binding. PMID- 10383437 TI - Binding studies with mutants of Zif268. Contribution of individual side chains to binding affinity and specificity in the Zif268 zinc finger-DNA complex. AB - The Zif268 zinc finger-DNA complex has served as a model system for understanding how Cys2His2 type zinc fingers recognize DNA. Structural studies of the Zif268 DNA complex revealed that residues at four positions in the alpha helix of each zinc finger play key roles in recognition, but there has been no information about the precise contributions of individual residues. Here we report the results of binding studies involving five mutants of Zif268 that have changes in the base-contacting residues of finger one. These studies let us evaluate the contributions that Arg18 (position -1 of the alpha helix), Asp20 (position 2), Glu21 (position 3), and Arg24 (position 6) make to the overall energy of DNA binding. Our results confirm the important role played by these arginines. By comparing the affinities of the wild type and mutant peptides for various sites, we also prove that Asp20 and Glu21 play important roles in determining binding site specificity. PMID- 10383438 TI - Eukaryotic molybdopterin synthase. Biochemical and molecular studies of Aspergillus nidulans cnxG and cnxH mutants. AB - We describe the primary structure of eukaryotic molybdopterin synthase small and large subunits and compare the sequences of the lower eukaryote, Aspergillus nidulans, and a higher eukaryote, Homo sapiens. Mutants in the A. nidulans cnxG (encoding small subunit) and cnxH (large subunit) genes have been analyzed at the biochemical and molecular level. Chlorate-sensitive mutants, all the result of amino acid substitutions, were shown to produce low levels of molybdopterin, and growth tests suggest that they have low levels of molybdoenzymes. In contrast, chlorate-resistant cnx strains have undetectable levels of molybdopterin, lack the ability to utilize nitrate or hypoxanthine as sole nitrogen sources, and are probably null mutations. Thus on the basis of chlorate toxicity, it is possible to distinguish between amino acid substitutions that permit a low level of molybdopterin production and those mutations that completely abolish molybdopterin synthesis, most likely reflecting molybdopterin synthase activity per se. Residues have been identified that are essential for function including the C-terminal Gly of the small subunit (CnxG), which is thought to be crucial for the sulfur transfer process during the formation of molybdopterin. Two independent alterations at residue Gly-148 in the large subunit, CnxH, result in temperature sensitivity suggesting that this residue resides in a region important for correct folding of the fungal protein. Many years ago it was proposed, from data showing that temperature-sensitive cnxH mutants had thermolabile nitrate reductase, that CnxH is an integral part of the molybdoenzyme nitrate reductase (MacDonald, D. W., and Cove, D. J. (1974) Eur. J. Biochem. 47, 107-110). Studies of temperature-sensitive cnxH mutants isolated in the course of this study do not support this hypothesis. Homologues of both molybdopterin synthase subunits are evident in diverse eukaryotic sources such as worm, rat, mouse, rice, and fruit fly as well as humans as discussed in this article. In contrast, molybdopterin synthase homologues are absent in the yeast Saccharomyces cerevisiae. Precursor Z and molybdopterin are undetectable in this organism nor do there appear to be homologues of molybdoenzymes. PMID- 10383439 TI - Receptor-mediated internalization is critical for the inhibition of the expression of growth hormone by somatostatin in the pituitary cell line AtT-20. AB - The inhibitory effect of the neuropeptide somatostatin on the expression of growth hormone was measured by quantitative polymerase chain reaction in the pituitary cell line AtT-20. We demonstrate that this effect is dependent on the internalization of somatostatin-receptor complexes and that it is totally independent from the peptide-induced inhibition of adenylate cyclase. Indeed, the inhibitory effect of the peptide on growth hormone mRNA levels was totally insensitive to pertussis toxin treatment but was totally abolished under conditions which block somatostatin receptor internalization. Comparative confocal microscopic imaging of fluorescent somatostatin sequestration and fluorescence immunolabeling of sst1, sst2A, and sst5 receptors suggests that sst2A is most probably responsible of the inhibitory effect of somatostatin on growth hormone expression. PMID- 10383440 TI - STAT3 activation in stromal/osteoblastic cells is required for induction of the receptor activator of NF-kappaB ligand and stimulation of osteoclastogenesis by gp130-utilizing cytokines or interleukin-1 but not 1,25-dihydroxyvitamin D3 or parathyroid hormone. AB - Interleukin (IL)-6-type cytokines stimulate osteoclastogenesis by activating gp130 in stromal/osteoblastic cells and may mediate some of the osteoclastogenic effects of other cytokines and hormones. To determine whether STAT3 is a downstream effector of gp130 in the osteoclast support function of stromal/osteoblastic cells and whether the gp130/STAT3 pathway is utilized by other osteoclastogenic agents, we conditionally expressed dominant negative (dn) STAT3 or dn-gp130 in a stromal/osteoblastic cell line (UAMS-32) that supports osteoclast formation. Expression of either dominant negative protein abolished osteoclast formation stimulated by IL-6 + soluble IL-6 receptor, oncostatin M, or IL-1 but not by parathyroid hormone or 1,25-dihydroxyvitamin D3. Because previous studies suggested that IL-6-type cytokines may stimulate osteoclastogenesis by inducing expression of the tumor necrosis factor-related protein, receptor activator of NF-kappaB ligand (RANKL), we conditionally expressed RANKL in UAMS 32 cells and found that this was sufficient to stimulate osteoclastogenesis. Moreover, dn-STAT3 blocked the ability of either IL-6 + soluble IL-6 receptor or oncostatin M to induce RANKL. These results establish that STAT3 is essential for gp130-mediated osteoclast formation and that the target of STAT3 during this process is induction of RANKL. In addition, this study demonstrates that activation of the gp130-STAT3 pathway in stromal/osteoblastic cells mediates the osteoclastogenic effects of IL-1, but not parathyroid hormone or 1, 25 dihydroxyvitamin D3. PMID- 10383441 TI - Identification of amino acids that modulate mannose phosphorylation of mouse DNase I, a secretory glycoprotein. AB - We have reported that bovine DNase I, a secretory glycoprotein, acquires mannose 6-phosphate residues on 12.6% of its Asn-linked oligosaccharides when expressed in COS-1 cells and that the extent of phosphorylation increases to 79.2% when lysines are placed at positions 27 and 74 of the mature protein (Nishikawa, A., Gregory, W. , Frenz, J., Cacia, J., and Kornfeld, S. (1997) J. Biol. Chem. 272, 19408-19412). We now demonstrate that murine DNase I, which contains Lys27 and Lys74, is phosphorylated only 20.9% when expressed in the same COS-1 cell system. This difference is mostly due to the absence of three residues present in bovine DNase I (Tyr54, Lys124, and Ser190) along with the presence of a valine at position 23 that is absent in the bovine species. We show that Val23 inhibits phosphorylation at the Asn18 glycosylation site, whereas Tyr54, Lys124, and Ser190 enhance phosphorylation at the Asn106 glycosylation site. Tyr54 and Ser190 are widely separated from each other and from Asn106 on the surface of DNase I, indicating that residues present over a broad area influence the interaction with UDP-GlcNAc:lysosomal enzyme N-acetylglucosamine-1-phosphotransferase, which is responsible for the formation of mannose 6-phosphate residues on lysosomal enzymes. PMID- 10383442 TI - Concurrent chaperone and protease activities of ClpAP and the requirement for the N-terminal ClpA ATP binding site for chaperone activity. AB - ClpA, a member of the Clp/Hsp100 family of ATPases, is both an ATP-dependent molecular chaperone and the regulatory component of ClpAP protease. We demonstrate that chaperone and protease activities occur concurrently in ClpAP complexes during a single round of RepA binding to ClpAP and ATP-dependent release. This result was substantiated with a ClpA mutant, ClpA(K220V), carrying an amino acid substitution in the N-terminal ATP binding site. ClpA(K220V) is unable to activate RepA, but the presence of ClpP or chemically inactivated ClpP restores its ability to activate RepA. The presence of ClpP simultaneously facilitates degradation of RepA. ClpP must remain bound to ClpA(K220V) for these effects, indicating that both chaperone and proteolytic activities of the mutant complex occur concurrently. ClpA(K220V) itself is able to form stable complexes with RepA in the presence of a poorly hydrolyzed ATP analog, adenosine 5'-O (thiotriphosphate), and to release RepA upon exchange of adenosine 5'-O (thiotriphosphate) with ATP. However, the released RepA is inactive in DNA binding, indicating that the N-terminal ATP binding site is essential for the chaperone activity of ClpA. Taken together, these results suggest that substrates bound to the complex of the proteolytic and ATPase components can be partitioned between release/reactivation and translocation/degradation. PMID- 10383443 TI - Intracellular reactive oxygen species mediate the linkage of Na+/K+-ATPase to hypertrophy and its marker genes in cardiac myocytes. AB - We showed before that in cardiac myocytes partial inhibition of Na+/K+-ATPase by nontoxic concentrations of ouabain causes hypertrophy and transcriptional regulations of growth-related marker genes through multiple Ca2+-dependent signal pathways many of which involve Ras and p42/44 mitogen-activated protein kinases. The aim of this work was to explore the roles of intracellular reactive oxygen species (ROS) in these ouabain-initiated pathways. Ouabain caused a rapid generation of ROS within the myocytes that was prevented by preexposure of cells to N-acetylcysteine (NAC) or vitamin E. These antioxidants also blocked or attenuated the following actions of ouabain: inductions of the genes of skeletal alpha-actin and atrial natriuretic factor, repression of the gene of the alpha3 subunit of Na+/K+-ATPase, activation of mitogen-activated protein kinases, activation of Ras-dependent protein synthesis, and activation of transcription factor NF-kappaB. Induction of c-fos and activation of AP-1 by ouabain were not sensitive to NAC. Ouabain-induced inhibition of active Rb+ uptake through Na+/K+ ATPase and the resulting rise in intracellular Ca2+ were also not prevented by NAC. A phorbol ester that also causes myocyte hypertrophy did not increase ROS generation, and its effects on marker genes and protein synthesis were not affected by NAC. We conclude the following: (a) ROS are essential second messengers within some but not all signal pathways that are activated by the effect of ouabain on Na+/K+-ATPase; (b) the ROS-dependent pathways are involved in ouabain-induced hypertrophy; (c) increased ROS generation is not a common response of the myocyte to all hypertrophic stimuli; and (d) it may be possible to dissociate the positive inotropic effect of ouabain from its growth-related effects by alteration of the redox state of the cardiac myocyte. PMID- 10383444 TI - Three-dimensional structure of guanylyl cyclase activating protein-2, a calcium sensitive modulator of photoreceptor guanylyl cyclases. AB - Guanylyl cyclase activating protein-2 (GCAP-2) is a Ca2+-sensitive regulator of phototransduction in retinal photoreceptor cells. GCAP-2 activates retinal guanylyl cyclases at low Ca2+ concentration (<100 nM) and inhibits them at high Ca2+ (>500 nM). The light-induced lowering of the Ca2+ level from approximately 500 nM in the dark to approximately 50 nM following illumination is known to play a key role in visual recovery and adaptation. We report here the three dimensional structure of unmyristoylated GCAP-2 with three bound Ca2+ ions as determined by nuclear magnetic resonance spectroscopy of recombinant, isotopically labeled protein. GCAP-2 contains four EF-hand motifs arranged in a compact tandem array like that seen previously in recoverin. The root mean square deviation of the main chain atoms in the EF-hand regions is 2.2 A in comparing the Ca2+-bound structures of GCAP-2 and recoverin. EF-1, as in recoverin, does not bind calcium because it contains a disabling Cys-Pro sequence. GCAP-2 differs from recoverin in that the calcium ion binds to EF-4 in addition to EF-2 and EF 3. A prominent exposed patch of hydrophobic residues formed by EF-1 and EF-2 (Leu24, Trp27, Phe31, Phe45, Phe48, Phe49, Tyr81, Val82, Leu85, and Leu89) may serve as a target-binding site for the transmission of calcium signals to guanylyl cyclase. PMID- 10383445 TI - Characterization of Ca2+-dependent phospholipase A2 activity during zebrafish embryogenesis. AB - We have developed a simple fluorescent assay for detection of phospholipase A2 (PLA2) activity in zebrafish embryos that utilizes a fluorescent phosphatidylcholine substrate. By using this assay in conjunction with selective PLA2 inhibitors and Western blot analysis, we identified the principal activity in zebrafish embryogenesis as characteristic of the Ca2+-dependent cytosolic PLA2 (cPLA2) subtype. Embryonic cPLA2 activity remained constant from the 1-cell stage until the onset of somitogenesis, at which time it increased sharply. This increase was preceded by the expression of a previously identified zebrafish cPLA2 homologue (Nalefski, E., Sultzman, L., Martin, D., Kriz, R., Towler, P., Knopf, J., and Clark, J. (1994) J. Biol. Chem. 269, 18239-18249). By using a quenched BODIPY-labeled phosphatidylcholine that fluoresces only upon cleavage by PLA2, lipase activity was visualized in the cells of living embryos where it localized to perinuclear membranes. PMID- 10383446 TI - Activation of the protein kinase Akt/PKB by the formation of E-cadherin-mediated cell-cell junctions. Evidence for the association of phosphatidylinositol 3 kinase with the E-cadherin adhesion complex. AB - E-cadherins are surface adhesion molecules localized at the level of adherens junctions, which play a major role in cell adhesiveness by mediating calcium dependent homophylic interactions at sites of cell-cell contacts. Recently, E cadherins have been also implicated in a number of biological processes, including cell growth and differentiation, cell recognition, and sorting during developmental morphogenesis, as well as in aggregation-dependent cell survival. As phosphatidylinositol (PI) 3-kinase and Akt play a critical role in survival pathways in response to both growth factors and extracellular stimuli, these observations prompted us to explore whether E-cadherins could affect intracellular molecules regulating the activity of the PI 3-kinase/Akt signaling cascade. Using Madin-Darby canine kidney cells as a model system, we show here that engagement of E-cadherins in homophylic calcium-dependent cell-cell interactions results in a rapid PI 3-kinase-dependent activation of Akt and the subsequent translocation of Akt to the nucleus. Moreover, we demonstrate that the activation of PI 3-kinase in response to cell-cell contact formation involves the phosphorylation of PI 3-kinase in tyrosine residues, and the concomitant recruitment of PI 3-kinase to E-cadherin-containing protein complexes. These findings indicate that E-cadherins can initiate outside-in signal transducing pathways that regulate the activity of PI 3-kinase and Akt, thus providing a novel molecular mechanism whereby the interaction among neighboring cells and their adhesion status may ultimately control the fate of epithelial cells. PMID- 10383447 TI - Hormone-dependent translocation of vitamin D receptors is linked to transactivation. AB - Vitamin D receptor (VDR) acts as a transcription factor mediating genomic actions of calcitriol. Our earlier studies suggested that calcitriol induces translocation of cytoplasmic VDR, but the physiologic relevance of this finding remained uncertain. Previous studies demonstrated that the activation function 2 domain (AF-2) plays an essential role in VDR transactivation. To elucidate hormone-dependent VDR translocation and its role, we constructed green fluorescent protein (GFP) chimeras with full-length VDR (VDR-GFP), AF-2-truncated VDR (AF-2del-VDR-GFP), and ligand-binding domain (LBD)-truncated VDR (LBDdel-VDR GFP). COS-7 cells were transiently transfected with these constructs. Western blot analysis, fluorescent microscopy, and transactivation assays showed that the generated chimeras are expressed and fluoresce and that VDR-GFP is transcriptionally active. After hormone treatment, cytoplasmic VDR-GFP translocated to the nucleus in a concentration-, time-, temperature-, and analog specific manner. Hormone dose-response relationships for translocation and for transactivation were similar. Truncation of LBD and truncation of AF-2 each abolished hormone-dependent translocation and transactivation. Our data confirm a hormone-dependent VDR translocation, demonstrate that an intact AF-2 domain is required for this translocation, and indicate that translocation is part of the receptor activation process. PMID- 10383448 TI - The role of intron structures in trans-splicing and cap 4 formation for the Leishmania spliced leader RNA. AB - A 39-nucleotide leader is trans-spliced onto all trypanosome nuclear mRNAs. The precursor spliced leader RNA was tested for trans-splicing function in vivo by mutating the intron. We report that in Leishmania tarentolae spliced leader RNA 5' modification is influenced by the primary sequence of stem-loop II, the Sm binding site, and the secondary structure of stem-loop III. The sequence of stem loop II was found to be important for cap 4 formation and splicing. As in Ascaris, mutagenesis of the bulge nucleotide in stem-loop II was detrimental to trans-splicing. Because restoration of the L. tarentolae stem-loop II structure was not sufficient to restore splicing, this result contrasts the findings in the kinetoplastid Leptomonas, where mutations that restored stem-loop II structure supported splicing. Methylation of the cap 4 structure and splicing was also dependent on both the Sm-binding site and the structure of stem-loop III and was inhibited by incomplete 3' end processing. The critical nature of the L. tarentolae Sm-binding site is consistent with its essential role in the Ascaris spliced leader RNA, whereas in Leptomonas mutation of the Sm-binding site and deletion of stem-loop III did not affect trans-splicing. A pathway for Leishmania spliced leader RNA processing and maturation is proposed. PMID- 10383449 TI - The human tumor necrosis factor (TNF) receptor-associated factor 1 gene (TRAF1) is up-regulated by cytokines of the TNF ligand family and modulates TNF-induced activation of NF-kappaB and c-Jun N-terminal kinase. AB - To understand how the TNF receptor-associated factor 1 (TRAF1) is transcriptionally regulated, in vitro DNA binding assays, promoter-reporter gene assays, and RNase protection assays were performed with the human TRAF1 gene. Binding of NF-kappaB to three of five putative binding sites within the human TRAF1 promoter was found in electrophoretic mobility shift assay studies, and analysis of TRAF1 gene promoter luciferase constructs confirmed the functional importance of these elements. Moreover, triggering of TNF-R1, CD40, and the interleukin-1 receptor resulted in transcription of the TRAF1 gene, whereas receptors that are not activators or only poor activators of NF-kappaB in HeLa cells failed to show a significant TRAF1 induction. Because it has been shown that members of the TRAF family are involved in activation of NF-kappaB and the c Jun N-terminal kinase (JNK) by the interleukin-1 receptor and members of the TNF receptor superfamily, a role of TRAF1 in receptor cross-talk and/or feedback regulation of activated receptor signaling complexes can be suggested. In fact, we found that TNF-induced activation of JNK is prolonged in transfectants overexpressing TRAF1, whereas overexpression of a deletion mutant of TRAF1 in which the N-terminal part had been replaced by the green fluorescent protein interfered with TNF-induced activation of NF-kappaB and JNK. PMID- 10383450 TI - Intracellular accumulation of secreted proteoglycans inhibits cationic lipid mediated gene transfer. Co-transfer of glycosaminoglycans to the nucleus. AB - Molecules secreted by potential target cells may interfere with cationic lipid mediated gene transfer. This has been studied using human lung fibroblasts and human epidermoid lung cancer cells. Secreted cell medium components caused a substantial decrease both in the uptake of cationic lipid-DNA complexes (2-4 fold) and in reporter gene expression (100-1000-fold). Metabolic labeling of the cell medium showed that especially [35S]sulfate-labeled macromolecules competed with DNA for binding to the cationic lipid. Release of DNA from the cationic lipid by cell medium components was demonstrated by an ethidium bromide intercalation assay. In the presence of the cationic lipid, the secreted macromolecules were internalized by the cells. By enzymatic digestions, it was shown that the competing macromolecules consist of chondroitin/dermatan sulfate and heparan sulfate proteoglycans and that the effects on transfection were mediated by the polyanionic glycosaminoglycan portion of the proteoglycan. Accordingly, pretreatment of cell medium with the polycationic peptide protamine sulfate abrogated the inhibitory effects on gene transfer. Fluorescence microscopy studies revealed that heparan sulfate, internalized as a complex with cationic lipids, accumulated in the cell nuclei. These results support the view that the lack of specificity of this type of gene transfer vehicle is a major hindrance to efficient and safe in vivo administration. PMID- 10383451 TI - Artemisinin, an endoperoxide antimalarial, disrupts the hemoglobin catabolism and heme detoxification systems in malarial parasite. AB - Endoperoxide antimalarials based on the ancient Chinese drug Qinghaosu (artemisinin) are currently our major hope in the fight against drug-resistant malaria. Rational drug design based on artemisinin and its analogues is slow as the mechanism of action of these antimalarials is not clear. Here we report that these drugs, at least in part, exert their effect by interfering with the plasmodial hemoglobin catabolic pathway and inhibition of heme polymerization. In an in vitro experiment we observed inhibition of digestive vacuole proteolytic activity of malarial parasite by artemisinin. These observations were further confirmed by ex vivo experiments showing accumulation of hemoglobin in the parasites treated with artemisinin, suggesting inhibition of hemoglobin degradation. We found artemisinin to be a potent inhibitor of heme polymerization activity mediated by Plasmodium yoelii lysates as well as Plasmodium falciparum histidine-rich protein II. Interaction of artemisinin with the purified malarial hemozoin in vitro resulted in the concentration-dependent breakdown of the malaria pigment. Our results presented here may explain the selective and rapid toxicity of these drugs on mature, hemozoin-containing, stages of malarial parasite. Since artemisinin and its analogues appear to have similar molecular targets as chloroquine despite having different structures, they can potentially bypass the quinoline resistance machinery of the malarial parasite, which causes sublethal accumulation of these drugs in resistant strains. PMID- 10383452 TI - Modifications of Igalpha and Igbeta expression as a function of B lineage differentiation. AB - Transcription of the mb1 and B29 genes is initiated when lymphoid progenitors enter the B cell differentiation pathway, and their transmembrane Igalpha and Igbeta products constitute essential signaling components of pre-B and B cell antigen receptors. We analyzed Igalpha/Igbeta biosynthesis, heterogeneity, and molecular interactions as a function of human B lineage differentiation in cell lines representative of the pro-B, pre-B, and B cell stages. All B lineage representatives produced a 36-kDa Igbeta form and three principal Igalpha forms, transient 33/40-kDa species and a mature 44-kDa glycoprotein. Deglycosylation revealed a major Igalpha core protein of 25 kDa and a minor 21-kDa Igalpha protein, apparently the product of an alternatively spliced mRNA. In pro-B cells, the Igalpha and Igbeta molecules existed primarily in separate unassembled pools, exhibited an immature glycosylation pattern, did not associate with surrogate light chain proteins, and were retained intracellularly. Their unanticipated association with the Lyn protein-tyrosine kinase nevertheless suggests functional potential for the Igalpha/Igbeta molecules in pro-B cells. Greater heterogeneity of the Igalpha and Igbeta molecules in pre-B and B cell lines was attributable to increased glycosylation complexity. Finally, the Igalpha/Igbeta heterodimers associated with fully assembled IgM molecules as a terminal event in B cell receptor assembly. PMID- 10383453 TI - Mutational alterations in the homotetrameric chaperone SecB that implicate the structure as dimer of dimers. AB - Variant forms of SecB with substitutions of aminoacyl residues in the region from 74 to 80 were analyzed with respect to their ability to bind a physiological ligand, precursor galactose-binding protein, and to their oligomeric states. SecBL75Q and SecBE77K are tetramers with affinity for ligand indistinguishable from that of the wild-type SecB, and thus the export defect exhibited by strains producing these variants must result from an effect on interactions between SecB and other components. SecBF74I is tetrameric but binds ligand with a lower affinity. Substitutions at positions 76, 78, and 80 cause a shift in the equilibrium so that the SecB tetramer dissociates into dimers. We conclude that the tetramer is a dimer of dimers and that the residues Cys76, Val78, and Gln80 must be involved either directly or indirectly in forming the interface between dimers. These variant species are defective in binding ligand; however, because their oligomeric state is altered no conclusion can be drawn concerning the direct role of these residues in ligand binding. PMID- 10383454 TI - IRAK-M is a novel member of the Pelle/interleukin-1 receptor-associated kinase (IRAK) family. AB - The interleukin-1 receptor-associated kinase (IRAK) was first described as a signal transducer for interleukin-1 (IL-1) and has later been implicated in signal transduction of other members of the Toll/IL-1 receptor family. We now report the identification and characterization of a novel IRAK-like molecule. In contrast to the ubiquitously expressed IRAK and IRAK-2, this new IRAK-like molecule is found mainly in cells of monomyeloic origin and is, therefore, designated IRAK-M. Although IRAK-M and IRAK-2 exhibit only a negligible autophosphorylation activity, they can reconstitute the IL-1 response in a 293 mutant cell line lacking IRAK. In addition, we show for the first time that members of the IRAK family are indispensable elements of lipopolysaccharide signal transduction. The discovery of IRAK-M adds another level of complexity to our understanding of signaling by members of the Toll/IL-1 receptor family. PMID- 10383456 TI - Nuclear localization of the 82-kDa form of human choline acetyltransferase. AB - Choline acetyltransferase is the enzyme catalyzing synthesis of the neurotransmitter acetylcholine in cholinergic neurons. In human, transcripts encoding two forms of the enzyme with apparent molecular masses of 69 and 82 kDa are found in brain and spinal cord; the 82-kDa form differs from the 69-kDa enzyme only in terms of a 118-amino acid extension on its amino terminus. Using green fluorescent protein-tagged choline acetyltransferase, we show that the 82 kDa enzyme is targeted to nuclei of cells, whereas the 69-kDa protein is found in cytoplasm. Expression of site-directed and deletion mutants of the 82-kDa isoform reveals that the extended amino terminus contains a nuclear localization signal in the first nine amino acids which targets the protein to nucleus. This represents the first report of a neurotransmitter-synthesizing enzyme that is localized to the cell nucleus. PMID- 10383455 TI - Tumor necrosis factor induces phosphorylation and translocation of BAD through a phosphatidylinositide-3-OH kinase-dependent pathway. AB - Tumor necrosis factor (TNF) induced the phosphorylation of BAD at serine 136 in HeLa cells under conditions that are not cytotoxic. BAD phosphorylation by TNF was dependent on phosphatidylinositide-3-OH kinase (PI3K) and was accompanied by the translocation of BAD from the mitochondria to the cytosol. Blocking the phosphorylation of BAD and its translocation to the cytosol with the PI3K inhibitor wortmannin activated caspase-3 and markedly potentiated the cytotoxicity of TNF. Transient transfection with a PI3K dominant negative mutant or a dominant negative mutant of the serine-threonine kinase Akt, the downstream target of PI3K and the enzyme that phosphorylates BAD, similarly potentiated the cytotoxicity of TNF. By contrast, transfection with a constitutively active Akt mutant protected against the cytotoxicity of TNF in the presence of wortmannin. Phosphorylation of BAD prevents its interaction with the antiapoptotic protein Bcl-XL. Transfection with a Bcl-XL expression vector protected against the cytotoxicity of TNF in the presence of wortmannin. The mechanism by which the inhibition of the phosphorylation of BAD is likely linked to the induction of lethal mitochondrial damage in TNF-intoxicated cells is discussed. PMID- 10383457 TI - Role of Sp1 in cAMP-dependent transcriptional regulation of the bovine CYP11A gene. AB - The pituitary peptide hormone ACTH regulates transcription of the cholesterol side chain cleavage cytochrome P450 (CYP11A) gene via cAMP and activation of cAMP dependent protein kinase. A G-rich sequence element conferring cAMP-dependent regulation has been found to reside within region -118 to -100 of the bovine CYP11A promoter. Previous studies have suggested that it binds a protein antigenically related to the transcription factor Sp1. We now report that the 118/-100 element binds both Sp1 and Sp3, members of the Sp family of transcription factors. We have made use of Drosophila SL2 cells, which lack endogenous Sp factors, to dissect the possible functional roles of Sp1, Sp3, and Sp4. All factors stimulated the activity of cotransfected reporter constructs in which the promoter of the bovine CYP11A gene regulates luciferase expression. Sp3 did not repress Sp1-dependent activation, as has previously been shown for other G-rich promoters. Mutation of the -118/-100 element of CYP11A abolished Sp1 mediated activation of a CYP11A reporter gene in SL2 cells as well as cAMP responsiveness in human H295R cells. Furthermore, cotransfection of SL2 cells with the catalytic subunit of cAMP-dependent protein kinase together with Sp1 and a CYP11A reporter construct enhanced Sp1-dependent activation of the reporter 4.2 fold, demonstrating that Sp1 confers cAMP responsiveness in these cells. Thus, we show that introduction of Sp1 alone in an Sp-negative cell such as SL2 is sufficient to achieve the cAMP-dependent regulation observed using the -118/-100 element of CYP11A in adrenocortical cells. PMID- 10383458 TI - Genetic deficiency of acylation stimulating protein (ASP(C3ades-Arg)) does not cause hyperapobetalipoproteinemia in mice. AB - The acylation stimulating protein (ASP) is a 76-amino acid peptide that has been proposed as a potent mediator of triglyceride synthesis and, when functionally impaired, as a major cause of hyperapobetalipoproteinemia (HyperapoB). Purification and sequence analysis of ASP from human sera have revealed that ASP is identical to the complement C3-derived activation peptide C3ades-Arg. Because C3 is the precursor for C3ades-Arg and therefore ASP, a deficiency in C3 would be predicted to result in a phenotype characteristic of HyperapoB. To test this hypothesis in vivo, the current study was undertaken in which ASP(C3ades-Arg) deficient mice were used as a model system. No significant differences were found in the triglyceride, cholesterol, or free fatty acid concentrations in the plasma of fasted normal and ASP(C3ades-Arg)-deficient animals. In addition, plasma lipoprotein analyses indicated that the very low density lipoprotein, low density lipoprotein, and high density lipoprotein cholesterol and triglyceride concentrations as well as the apolipoprotein B-48 and B-100 levels were not significantly different in the plasma of ASP(C3ades-Arg)-deficient and wild type mice. Furthermore, when challenged with an oral fat load, the ASP(C3ades-Arg) deficient mice showed no impaired ability to clear triglycerides and free fatty acids from their circulation when compared with their wild-type littermates. Collectively, these results indicate that ASP(C3ades-Arg) deficiency does not cause HyperapoB in mice and that the physiological importance of impaired ASP(C3ades-Arg) function as a cause of hyperapobetalipoproteinemia needs to be reevaluated. PMID- 10383459 TI - Identification of three major phosphorylation sites within HIV-1 capsid. Role of phosphorylation during the early steps of infection. AB - We previously reported the presence of two cellular serine/threonine protein kinases incorporated in human immunodeficiency virus type 1 (HIV-1) particles. One protein kinase is MAPK ERK2 (mitogen-activated protein kinase), whereas the other one, a 53-kDa protein, still needs to be identified. Furthermore, we demonstrated that the capsid protein CAp24 is phosphorylated by one of those two virion-associated protein kinases (Cartier, C., Deckert, M., Grangeasse, C., Trauger, R., Jensen, F., Bernard, A., Cozzone, A., Desgranges, C., and Boyer, V. (1997) J. Virol. 71, 4832-4837). In this study, we showed that CAp24 is not a direct substrate of MAPK ERK2. Moreover, using site-directed mutagenesis of each of the 9 serine residues of CAp24, we demonstrated the phosphorylation of 3 serine residues (Ser-109, Ser-149, and Ser-178) in the CAp24. Substitution of each serine residue did not affect viral budding, nor viral structure. By contrast, substitution of Ser-109, Ser-149, or Ser-178 affects viral infectivity by preventing the reverse transcription process to be completely achieved. Our results suggest that CAp24 serine phosphorylation is essential for viral uncoating process. PMID- 10383460 TI - Ubc9 interacts with the androgen receptor and activates receptor-dependent transcription. AB - Ubc9, a homologue of the class E2 ubiquitin-conjugating enzymes, has recently been shown to catalyze conjugation of a small ubiquitin-like molecule-1 (SUMO-1) to a variety of target proteins. SUMO-1 modifications have been implicated in the targeting of proteins to the nuclear envelope and certain intranuclear structures and in converting proteins resistant to ubiquitin-mediated degradation. In the present work, we find that Ubc9 interacts with the androgen receptor (AR), a member of the steroid receptor family of ligand-activated transcription factors. In transiently transfected COS-1 cells, AR-dependent but not basal transcription is enhanced by the coexpression of Ubc9. The N-terminal half of the AR hinge region containing the C-terminal part of the bipartite nuclear localization signal is essential for the interaction with Ubc9. Deletion of this part of the nuclear localization signal, which does not completely prevent the transfer of AR to the nucleus, abolishes the AR-Ubc9 interaction and attenuates the transcriptional response to cotransfected Ubc9. The C93S substitution of Ubc9, which prevents SUMO-1 conjugation by abrogating the formation of a thiolester bond between SUMO-1 and Ubc9, does not influence the capability of Ubc9 to stimulate AR-dependent transactivation, implying that Ubc9 is able to act as an AR coregulator in a fashion independent of its ability to catalyze SUMO-1 conjugation. PMID- 10383461 TI - Critical role of cAMP response element binding protein expression in hypoxia elicited induction of epithelial tumor necrosis factor-alpha. AB - Tissue hypoxia is intimately associated with a number of chronic inflammatory conditions of the intestine. In this study, we investigated the impact of hypoxia on the expression of a panel of inflammatory mediators by intestinal epithelia. Initial experiments revealed that epithelial (T84 cell) exposure to ambient hypoxia evoked a time-dependent induction of the proinflammatory markers tumor necrosis factor-alpha (TNF-alpha), interleukin-8 (IL-8), and major histocompatibility complex (MHC) class II (37 +/- 6.1-, 7 +/- 0.8-, and 9 +/- 0.9 fold increase over normoxia, respectively, each p < 0.01). Since the gene regulatory elements for each of these molecules contains an NF-kappaB binding domain, we investigated the influence of hypoxia on NF-kappaB activation. Cellular hypoxia induced a time-dependent increase in nuclear p65, suggesting a dominant role for NF-kappaB in hypoxia-elicited induction of proinflammatory gene products. Further work, however, revealed that hypoxia does not influence epithelial intercellular adhesion molecule 1 (ICAM-1) or MHC class I, the promoters of which also contain NF-kappaB binding domains, suggesting differential responses to hypoxia. Importantly, the genes for TNF-alpha, IL-8, and MHC class II, but not ICAM-1 or MHC class I, contain cyclic AMP response element (CRE) consensus motifs. Thus, we examined the role of cAMP in the hypoxia elicited phenotype. Hypoxia diminished CRE binding protein (CREB) expression. In parallel, T84 cell cAMP was diminished by hypoxia (83 +/- 13.2% decrease, p < 0.001), and pharmacologic inhibition of protein kinase A induced TNF-alpha and protein release (9 +/- 3.9-fold increase). Addback of cAMP resulted in reversal of hypoxia-elicited TNF-alpha release (86 +/- 3.2% inhibition with 3 mM 8-bromo cAMP). Furthermore, overexpression of CREB but not mutated CREB by retroviral mediated gene transfer reversed hypoxia-elicited induction of TNF-alpha defining a causal relationship between hypoxia-elicited CREB reduction and TNF-alpha induction. Such data indicate a prominent role for CREB in the hypoxia-elicited epithelial phenotype and implicate intracellular cAMP as an important second messenger in differential induction of proinflammatory mediators. PMID- 10383462 TI - Two cytoplasmic loops of the glucagon receptor are required to elevate cAMP or intracellular calcium. AB - The glucagon receptor is a member of a distinct class of G protein-coupled receptors (GPCRs) sharing little amino acid sequence homology with the larger rhodopsin-like GPCR family. To identify the components of the glucagon receptor necessary for G-protein coupling, we replaced sequentially all or part of each intracellular loop (i1, i2, and i3) and the C-terminal tail of the glucagon receptor with the 11 amino acids comprising the first intracellular loop of the D4 dopamine receptor. When expressed in transiently transfected COS-1 cells, the mutant receptors fell into two different groups with respect to hormone-mediated signaling. The first group included the loop i1 mutants, which bound glucagon and signaled normally. The second group comprised the loop i2 and i3 chimeras, which caused no detectable adenylyl cyclase activation in COS-1 cells. However, when expressed in HEK 293T cells, the loop i2 or i3 chimeras caused very small glucagon-mediated increases in cAMP levels and intracellular calcium concentrations, with EC50 values nearly 100-fold higher than those measured for wild-type receptor. Replacement of both loops i2 and i3 simultaneously was required to completely abolish G protein signaling as measured by both cAMP accumulation and calcium flux assays. These results show that the i2 and i3 loops play a role in glucagon receptor signaling, consistent with recent models for the mechanism of activation of G proteins by rhodopsin-like GPCRs. PMID- 10383463 TI - Isolation and biochemical characterization of the human Dkk-1 homologue, a novel inhibitor of mammalian Wnt signaling. AB - In an effort to isolate novel growth factors, we identified a human protein, designated Sk, that co-eluted with Neuregulin during chromatographic separation of conditioned medium from the SK-LMS-1 human leiomyosarcoma cell line. Degenerate oligonucleotides based on amino-terminal sequence analysis of the purified protein were used to isolate the corresponding cDNA from a library generated from this cell line. Sk is a novel 266-amino acid protein that contains a signal peptide sequence and two cysteine-rich domains with no similarity to other known growth factors. A single major 2-kilobase transcript was expressed in several embryonic tissues. Transfection of mammalian cells demonstrated that the protein was secreted and expressed as a doublet of approximately 35 kDa. In vitro translation and endoglycosylase analysis indicated that this doublet, which was also observed in cells expressing the endogenous protein, arises from posttranslational modification. A search of the GenBankTM data base revealed a match of Sk with Dkk-1, which is a novel secreted protein required for head induction in amphibian embryos and a potent Wnt inhibitor. When coexpressed with Wnt-2 in NIH3T3 cells, human Sk/Dkk-1 caused reversion of Wnt-2 induced morphological alterations and inhibited the Wnt-2 induced increase in uncomplexed beta-catenin levels. These results provide biochemical evidence that human Sk/Dkk 1 antagonizes Wnt signaling upstream of its effect on beta-catenin regulation. PMID- 10383464 TI - Arcadlin is a neural activity-regulated cadherin involved in long term potentiation. AB - Neural activity results in long term changes that underlie synaptic plasticity. To examine the molecular basis of activity-dependent plasticity, we have used differential cloning techniques to identify genes that are rapidly induced in brain neurons by synaptic activity. Here, we identify a novel cadherin molecule Arcadlin (activity-regulated cadherin-like protein). arcadlin mRNA is rapidly and transiently induced in hippocampal granule cells by seizures and by N-methyl-D aspartate-dependent synaptic activity in long term potentiation. The extracellular domain of Arcadlin is most homologous to protocadherin-8; however, the cytoplasmic region is distinct from that of any cadherin family member. Arcadlin protein is expressed at the synapses and shows a homophilic binding activity in a Ca2+-dependent manner. Furthermore, application of Arcadlin antibody reduces excitatory postsynaptic potential amplitude and blocks long term potentiation in hippocampal slices. Its close homology with cadherins, its rapid inducibility by neural activity, and its involvement in synaptic transmission suggest that Arcadlin may play an important role in activity-induced synaptic reorganization underlying long term memory. PMID- 10383465 TI - Scanning cysteine accessibility of EmrE, an H+-coupled multidrug transporter from Escherichia coli, reveals a hydrophobic pathway for solutes. AB - EmrE is a 12-kDa Escherichia coli multidrug transporter that confers resistance to a wide variety of toxic reagents by actively removing them in exchange for hydrogen ions. The three native Cys residues in EmrE are inaccessible to N ethylmaleimide (NEM) and a series of other sulfhydryls. In addition, each of the three residues can be replaced with Ser without significant loss of activity. A protein without all the three Cys residues (Cys-less) has been generated and shown to be functional. Using this Cys-less protein, we have now generated a series of 48 single Cys replacements throughout the protein. The majority of them (43) show transport activity as judged from the ability of the mutant proteins to confer resistance against toxic compounds and from in vitro analysis of their activity in proteoliposomes. Here we describe the use of these mutants to study the accessibility to NEM, a membrane permeant sulfhydryl reagent. The study has been done systematically so that in one transmembrane segment (TMS2) each single residue was replaced. In each of the other three transmembrane segments, at least four residues covering one turn of the helix were replaced. The results show that although the residues in putative hydrophilic loops readily react with NEM, none of the residues in putative transmembrane domains are accessible to the reagent. The results imply very tight packing of the protein without any continuous aqueous domain. Based on the findings described in this work, we conclude that in EmrE the substrates are translocated through a hydrophobic pathway. PMID- 10383466 TI - Identification and molecular characterization of m3 muscarinic receptor dimers. AB - Several studies suggest, but do not prove directly, that muscarinic receptors may be able to form dimeric or oligomeric arrays. To address this issue in a more direct fashion, we designed a series of biochemical experiments using a modified version of the rat m3 muscarinic receptor (referred to as m3') as a model system. When membrane lysates prepared from m3' receptor-expressing COS-7 cells were subjected to Western blot analysis under non-reducing conditions, several immunoreactive species were observed corresponding in size to putative receptor monomers, dimers, and oligomers. However, under reducing conditions, the monomeric receptor species represented the only detectable immunoreactive protein, consistent with the presence of disulfide-linked m3 receptor complexes. Similar results were obtained when native m3 muscarinic receptors present in rat brain membranes were analyzed. Control experiments carried out in the presence of high concentrations of the SH group alkylating agent, N-ethylmaleimide, suggested that disulfide bond formation did not occur artifactually during the preparation of cell lysates. The formation of m3' receptor dimers/multimers was confirmed in coimmunoprecipitation studies using differentially epitope-tagged m3' receptor constructs. In addition, these studies showed that m3' receptors were also able to form non-covalently associated receptor dimers and that m3' receptor dimer formation was receptor subtype-specific. Immunological studies also demonstrated that m3' receptor dimers/multimers were abundantly expressed on the cell surface. Site-directed mutagenesis studies indicated that two conserved extracellular Cys residues (Cys-140 and Cys-220) play key roles in the formation of disulfide linked m3' receptor dimers. These results provide the first direct evidence for the existence of muscarinic receptor dimers and highlight the specificity and molecular diversity of G protein-coupled receptor dimerization/oligomerization. PMID- 10383467 TI - Transcriptional repression from the c-myc P2 promoter by the zinc finger protein ZF87/MAZ. AB - ZF87/MAZ is a zinc finger-containing transcription factor that was cloned based on its ability to bind to a site within the c-myc P2 promoter. However, its role in the control of c-myc transcription has not yet been well established. Here we have analyzed the effect of ZF87/MAZ overexpression on transcription from the murine c-myc P2 promoter. It was found that when overexpressed in COS cells, ZF87/MAZ significantly represses transcription from P2. The repression is mediated through the ME1a2 element, located at position -86 relative to the P2 transcriptional start site, and is not mediated through either the E2F or the ME1a1 sites. ZF87/MAZ functions as a true transcriptional repressor since it can repress transcription independently of the c-myc promoter, as part of a fusion with the GAL4 protein. The repressive domain within ZF87/MAZ is located in the amino-terminal half of the protein, a region rich in proline and alanine residues. ZF87/MAZ therefore shares features (i.e. a Pro/Ala-rich region) with those of known transcriptional repressor proteins. PMID- 10383468 TI - Class II MHC quantitative binding motifs derived from a large molecular database with a versatile iterative stepwise discriminant analysis meta-algorithm. AB - MOTIVATION: The identification of T-cell epitopes can be crucial for vaccine development. An epitope is a peptide segment that binds to both a T-cell receptor and a major histocompatibility complex (MHC) molecule. Predicting which peptide segments bind MHC molecules is the first step in epitope prediction. RESULTS: An iterative stepwise discriminant analysis meta-algorithm explores a large molecular database to derive quantitative motifs for peptide binding. The applications presented here demonstrate the algorithm's versatility by producing four closely related models for HLA-DR1. Two models use an expert initial estimate and two do not; two models use amino acid residues as the only predictors and two use amino acid groupings as additional predictors. Each model correctly classifies >90% of the peptides in the database. AVAILABILITY: Software is available commercially; data are free over the Internet. PMID- 10383469 TI - Fast evaluation of internal loops in RNA secondary structure prediction. AB - MOTIVATION: Though not as abundant in known biological processes as proteins, RNA molecules serve as more than mere intermediaries between DNA and proteins. Research in the last 15 years demonstrates that RNA molecules serve in many roles, including catalysis. Furthermore, RNA secondary structure prediction based on free energy rules for stacking and loop formation remains one of the few major breakthroughs in the field of structure prediction, as minimum free energy structures and related quantities can be computed with full mathematical rigor. However, with the current energy parameters, the algorithms used hitherto suffer the disadvantage of either employing heuristics that risk (though highly unlikely) missing the optimal structure or becoming prohibitively time consuming for moderate to large sequences. RESULTS: We present a new method to evaluate internal loops utilizing currently used energy rules. This method reduces the time complexity of this part of the structure prediction from O(n4) to O(n3), thus reducing the overall complexity to O(n3). Even when the size of evaluated internal loops is bounded by k (a commonly used heuristic), the method presented has a competitive edge by reducing the time complexity of internal loop evaluation from O(k2n2) to O(kn2). The method also applies to the calculation of the equilibrium partition function. AVAILABILITY: Source code for an RNA secondary structure prediction program implementing this method is available at ftp://www.ibc.wustl.edu/pub/zuker/zuker .tar.Z PMID- 10383470 TI - RNA secondary structure prediction using stochastic context-free grammars and evolutionary history. AB - MOTIVATION: Many computerized methods for RNA secondary structure prediction have been developed. Few of these methods, however, employ an evolutionary model, thus relevant information is often left out from the structure determination. This paper introduces a method which incorporates evolutionary history into RNA secondary structure prediction. The method reported here is based on stochastic context-free grammars (SCFGs) to give a prior probability distribution of structures. RESULTS: The phylogenetic tree relating the sequences can be found by maximum likelihood (ML) estimation from the model introduced here. The tree is shown to reveal information about the structure, due to mutation patterns. The inclusion of a prior distribution of RNA structures ensures good structure predictions even for a small number of related sequences. Prediction is carried out using maximum a posteriori estimation (MAP) estimation in a Bayesian approach. For small sequence sets, the method performs very well compared to current automated methods. PMID- 10383472 TI - Blocks+: a non-redundant database of protein alignment blocks derived from multiple compilations. AB - MOTIVATION: As databanks grow, sequence classification and prediction of function by searching protein family databases becomes increasingly valuable. The original Blocks Database, which contains ungapped multiple alignments for families documented in Prosite, can be searched to classify new sequences. However, Prosite is incomplete, and families from other databases are now available to expand coverage of the Blocks Database. RESULTS: To take advantage of protein family information present in several existing compilations, we have used five databases to construct Blocks+, a unified database that is built on the PROTOMAT/BLOSUM scoring model and that can be searched using a single algorithm for consistent sequence classification. The LAMA blocks-versus-blocks searching program identifies overlapping protein families, making possible a non-redundant hierarchical compilation. Blocks+ consists of all blocks derived from PROSITE, blocks from Prints not present in PROSITE, blocks from Pfam-A not present in PROSITE or Prints, and so on for ProDom and Domo, for a total of 1995 protein families represented by 8909 blocks, doubling the coverage of the original Blocks Database. A challenge for any procedure aimed at non-redundancy is to retain related but distinct families while discarding those that are duplicates. We illustrate how using multiple compilations can minimize this potential problem by examining the SNF2 family of ATPases, which is detectably similar to distinct families of helicases and ATPases. AVAILABILITY: http://blocks.fhcrc.org/ PMID- 10383471 TI - Family pairwise search with embedded motif models. AB - MOTIVATION: Statistical models of protein families, such as position-specific scoring matrices, profiles and hidden Markov models, have been used effectively to find remote homologs when given a set of known protein family members. Unfortunately, training these models typically requires a relatively large set of training sequences. Recent work (Grundy, J. Comput. Biol., 5,<479-492, 1998) has shown that, when only a few family members are known, several theoretically justified statistical modeling techniques fail to provide homology detection performance on a par with Family Pairwise Search (FPS), an algorithm that combines scores from a pairwise sequence similarity algorithm such as BLAST. RESULTS: The present paper provides a model-based algorithm that improves FPS by incorporating hybrid motif-based models of the form generated by Cobbler (Henikoff and Henikoff, Protein Sci., 6, 698-705, 1997). For the 73 protein families investigated here, this cobbled FPS algorithm provides better homology detection performance than either Cobbler or FPS alone. This improvement is maintained when BLAST is replaced with the full Smith-Waterman algorithm. AVAILABILITY: http://fps.sdsc.edu PMID- 10383473 TI - A comparison of sequence and structure protein domain families as a basis for structural genomics. AB - MOTIVATION: Protein families can be defined based on structure or sequence similarity. We wanted to compare two protein family databases, one based on structural and one on sequence similarity, to investigate to what extent they overlap, the similarity in definition of corresponding families, and to create a list of large protein families with unknown structure as a resource for structural genomics. We also wanted to increase the sensitivity of fold assignment by exploiting protein family HMMs. RESULTS: We compared Pfam, a protein family database based on sequence similarity, to Scop, which is based on structural similarity. We found that 70% of the Scop families exist in Pfam while 57% of the Pfam families exist in Scop. Most families that occur in both databases correspond well to each other, but in some cases they are different. Such cases highlight situations in which structure and sequence approaches differ significantly. The comparison enabled us to compile a list of the largest families that do not occur in Scop; these are suitable targets for structure prediction and determination, and may be useful to guide projects in structural genomics. It can be noted that 13 out of the 20 largest protein families without a known structure are likely transmembrane proteins. We also exploited Pfam to increase the sensitivity of detecting homologs of proteins with known structure, by comparing query sequences to Pfam HMMs that correspond to Scop families. For SWISSPROT+TREMBL, this yielded an increase in fold assignment from 31% to 42% compared to using FASTA only. This method assigned a structure to 22% of the proteins in Saccharomyces cerevisiae, 24% in Escherichia coli, and 16% in Methanococcus jannaschii. PMID- 10383474 TI - Clustering of non-polar contacts in proteins. AB - MOTIVATION: Hydrophobic or non-polar contacts in proteins are important for protein folding, protein stability and protein-protein interactions. In particular, in the interior of a protein, in the hydrophobic core, a large number of such contacts are found. The residues involved in these contacts often form a tightly packed cluster of atoms. It is useful for the understanding of protein structure to be able to identify and analyse such clusters. RESULTS: Tools for hierarchical cluster analysis of non-polar contacts in proteins are described. These tools allow for efficient identification of clusters of non-polar interactions in proteins, both internal clusters and clusters involved in protein protein contacts. The non-polar contacts are represented by a dendrogram structure, which is a simple approach for flexible identification of clusters by visual inspection. The tools are demonstrated on the structure of crambin, the structure of the complex between human growth hormone and the human growth hormone binding protein, and a pair of lipase/esterase structures. AVAILABILITY: On request from the author. PMID- 10383475 TI - An ontology for bioinformatics applications. AB - MOTIVATION: An ontology of biological terminology provides a model of biological concepts that can be used to form a semantic framework for many data storage, retrieval and analysis tasks. Such a semantic framework could be used to underpin a range of important bioinformatics tasks, such as the querying of heterogeneous bioinformatics sources or the systematic annotation of experimental results. RESULTS: This paper provides an overview of an ontology [the Transparent Access to Multiple Biological Information Sources (TAMBIS) ontology or TaO] that describes a wide range of bioinformatics concepts. The present paper describes the mechanisms used for delivering the ontology and discusses the ontology's design and organization, which are crucial for maintaining the coherence of a large collection of concepts and their relationships. AVAILABILITY: The TAMBIS system, which uses a subset of the TaO described here, is accessible over the Web via http://img.cs.man.ac.uk/tambis (although in the first instance, we will use a password mechanism to limit the load on our server). The complete model is also available on the Web at the above URL. PMID- 10383476 TI - Protein analyst--a distributed object environment for protein sequence and structure analysis. AB - SUMMARY: Protein Analyst is a flexible tool for the analysis of protein sequences with emphasis on the integration of sequence and structural information. AVAILABILITY: The software will be available from the Oxford Molecular Biolib web site (http://www. oxmol.co.uk/biolib) and will be free to the academic research community. PMID- 10383477 TI - BLAST PRINTS--alternative perspectives on sequence similarity. AB - SUMMARY: An implementation of BLAST for searching the PRINTS database is presented. The interface allows submission of either protein or DNA queries, and returns the familiar form of output, but modified by means of direct links both to the familial discriminators in PRINTS and to fingerprint profile visualization software. The server thus couples the rapidity of BLAST searching with the sensitivity of fingerprint diagnoses, providing alternative perspectives on a given query. AVAILABILITY: http://www.biochem.ucl. ac.uk/cgi bin/wright/printsBLAST.cgi PMID- 10383478 TI - SECOST: sequence-conformation-structure database for amino acid residues in proteins. AB - The sequence-conformation-structure database for amino acid residues contains information on 114 828 individual residues derived from the spatial structures of 473 high-quality non-homologous proteins. The information in the database is obtained using a variety of different methods and can be used in various protein modeling applications. PMID- 10383479 TI - A review of nutrients and botanicals in the integrative management of cognitive dysfunction. AB - Dementias and other severe cognitive dysfunction states pose a daunting challenge to existing medical management strategies. An integrative, early intervention approach seems warranted. Whereas, allopathic treatment options are highly limited, nutritional and botanical therapies are available which have proven degrees of efficacy and generally favorable benefit-to-risk profiles. This review covers five such therapies: phosphatidylserine (PS), acetyl-l-carnitine (ALC), vinpocetine, Ginkgo biloba extract (GbE), and Bacopa monniera (Bacopa). PS is a phospholipid enriched in the brain, validated through double-blind trials for improving memory, learning, concentration, word recall, and mood in middle-aged and elderly subjects with dementia or age-related cognitive decline. PS has an excellent benefit-to-risk profile. ALC is an energizer and metabolic cofactor which also benefits various cognitive functions in the middle-aged and elderly, but with a slightly less favorable benefit-to-risk profile. Vinpocetine, found in the lesser periwinkle Vinca minor, is an excellent vasodilator and cerebral metabolic enhancer with proven benefits for vascular-based cognitive dysfunction. Two meta-analyses of GbE demonstrate the best preparations offer limited benefits for vascular insufficiencies and even more limited benefits for Alzheimer's, while "commodity" GbE products offer little benefit, if any at all. GbE (and probably also vinpocetine) is incompatible with blood-thinning drugs. Bacopa is an Ayurvedic botanical with apparent anti-anxiety, anti-fatigue, and memory strengthening effects. These five substances offer interesting contributions to a personalized approach for restoring cognitive function, perhaps eventually in conjunction with the judicious application of growth factors. PMID- 10383480 TI - An ecologic study of dietary links to prostate cancer. AB - BACKGROUND: The etiology of prostate cancer has not been fully resolved in the scientific and medical literature, although the non-fat portion of milk and calcium are emerging as leading dietary risk factors, with lycopene (found in tomatoes) and vitamin D apparently being risk reduction factors. METHODS: The ecologic (multi-country statistical) approach is used to study dietary links to prostate cancer. Mortality data from 1986 for various age groups in 41 countries are compared with national consumer macronutrient supply values for 1983 and tomato supply values for 1985. RESULTS: For 28 countries with more than five Kcal/day of tomatoes in the consumer supply, a linear combination of non-fat milk (risk factor) and tomatoes (risk reduction factor) was found to have the highest statistical association with prostate cancer mortality rates for men over the age of 35, with the Pearson regression coefficient (R2) for those aged 65-74 years = 0.67 and p < 0.001. For the 13 countries with fewer than six Kcal/day of tomatoes, non-fat milk had the highest association (R2 = 0.92, p < 0.001 for men aged 65-74 years). For 41 countries combined, the non-fat portion of milk had the highest association with prostate cancer mortality rates (R2 = 0.73, p < 0.001 for men aged 65-74 years). CONCLUSIONS: These results support the results of several cohort studies which found the non-fat portion of milk to have the highest association with prostate cancer, likely due to the calcium, and tomatoes to reduce the risk of prostate cancer, most likely due to lycopene. PMID- 10383481 TI - Plant sterols and sterolins: a review of their immune-modulating properties. AB - Beta-sitosterol (BSS) and its glycoside (BSSG) are sterol molecules which are synthesized by plants. When humans eat plant foods phytosterols are ingested, and are found in the serum and tissues of healthy individuals, but at concentrations orders of magnitude lower than endogenous cholesterol. Epidemiological studies have correlated a reduced risk of numerous diseases with a diet high in fruits and vegetables, and have concluded that specific molecules, including b-carotene, tocopherols, vitamin C, and flavonoids, confer some of this protective benefit. However, these epidemiologic studies have not examined the potential effect that phytosterols ingested with fruits and vegetables might have on disease risk reduction. In animals, BSS and BSSG have been shown to exhibit anti-inflammatory, anti-neoplastic, anti-pyretic, and immune-modulating activity. A proprietary BSS:BSSG mixture has demonstrated promising results in a number of studies, including in vitro studies, animal models, and human clinical trials. This phytosterol complex seems to target specific T-helper lymphocytes, the Th1 and Th2 cells, helping normalize their functioning and resulting in improved T lymphocyte and natural killer cell activity. A dampening effect on overactive antibody responses has also been seen, as well as normalization of the DHEA:cortisol ratio. The re-establishment of these immune parameters may be of help in numerous disease processes relating to chronic immune-mediated abnormalities, including chronic viral infections, tuberculosis, rheumatoid arthritis, allergies, cancer, and auto-immune diseases. PMID- 10383482 TI - A review of plants used in the treatment of liver disease: part two. AB - Botanical medicines have been used traditionally by herbalists and indigenous healers worldwide for the prevention and treatment of liver disease. Clinical research in this century has confirmed the efficacy of several plants in the treatment of liver disease, while basic scientific research has uncovered the mechanisms by which some plants provide their therapeutic effects. This article is Part Two in a review of botanicals used in the treatment of liver disease. Curcuma longa (turmeric), Camellia sinensis (green tea), and Glycyrrhiza glabra (licorice) are reviewed in this installment. Silybum marianum (milk thistle) and Picrorhiza kurroa (kutkin) were reviewed in Part One. PMID- 10383483 TI - Hypericum. PMID- 10383484 TI - Glucosamine sulfate. AB - Glucosamine sulfate's role in halting or reversing joint degeneration appears to be directly due to its ability to act as an essential substrate for, and to stimulate the biosynthesis of, the glycosaminoglycans and the hyaluronic acid backbone needed for the formation of the proteoglycans found in the structural matrix of joints. Successful treatment of osteoarthritis must effectively control pain and should slow down or reverse the progression of the degeneration. Biochemical and pharmacological data combined with animal and human studies demonstrate that glucosamine sulfate is capable of satisfying both of these criteria. PMID- 10383485 TI - Synaptic and extrasynaptic roles of glutamate in the mammalian hippocampus. PMID- 10383486 TI - Oestrogen attenuates post-exercise myeloperoxidase activity in skeletal muscle of male rats. AB - The effects of 2 weeks of oestrogen (40 microg kg BW-1 beta-estradiol 3-benzoate) injection on 24 h post-exercise myeloperoxidase (MPO) activities were determined in plantaris and soleus muscles and liver of sexually mature male and female rats. The treadmill running protocol (45-60 min, at 28 m min-1, 15% grade) induced significant elevations in muscle MPO activities 24 h post-exercise in male rats, while prior oestrogen administration to male rats eliminated the post exercise elevations in muscle MPO activities. Female rats experienced no significant post-exercise elevations in muscle MPO activities. Hence oestrogen administration to male rats attenuated post-exercise muscle MPO activities to levels found in female animals. Liver MPO activities were not significantly affected by exercise, gender or oestrogen administration. Oestrogen may be a factor in diminishing 24 h post-exercise skeletal muscle leukocyte infiltration and inflammatory response in both male and female muscle. PMID- 10383487 TI - Impact of testosterone and oestradiol on region specificity of skeletal muscle ATP, creatine phosphokinase and myokinase in male and female Wistar rats. AB - The main aim of the present study was to test the hypothesis that skeletal muscle ATP concentration, creatine phosphokinase and myokinase enzyme activities are stimulated by the sex steroids in both male and female rats (animals were not subjected to any kind of exercise or any training). To test the hypothesis healthy mature (90-120 days old, weighing about 160-180 g) male and female rats were gonadectomized. Gonadectomized male and female rats were administered with testosterone (Sigma Chemical, St Louis, MO, USA) at a dose of 100 microg (100 g body weight)-1 day-1 for males and 5 microg (100 g body weight)-1 day-1 for females for 30 days from day 31 post-castration onwards; and oestradiol at a dose of 5 microg (100 g body weight)-1 day-1 for 30 days from day 31 post-castration onwards for both males and females (17beta oestradiol, Sigma Chemical Company, St Louis, MO, USA). The ATP content, creatine phosphokinase and myokinase enzyme activities of skeletal muscles were significantly higher than that of skeletal muscles of female control rats. Gonadectomy resulted in a significant decrease in ATP content and creatine phosphokinase myokinase enzyme activities in both male and female rats. Testosterone treatment to gonadectomized male rats brought back the parameters to normalcy whereas the same to the female rats enhanced the enzyme activities and ATP contents to the level of control male rats. Oestradiol treatment to castrated male rats did not bring about any significant alterations whereas the same in gonadectomized female rats brought them back to normalcy. Therefore from the present study it is concluded that testosterone is effective in both males and females whereas oestradiol was effective only in the females in enhancing skeletal muscle energy metabolism. PMID- 10383488 TI - Creatine uptake in isolated soleus muscle: kinetics and dependence on sodium, but not on insulin. AB - The increased use of creatine by athletes as a dietary supplement to improve their physical performance assumes that increased serum creatine levels will increase intracellular skeletal muscle creatine. Despite this common assumption, skeletal muscle creatine uptake awaits full characterization. Consequently, we have investigated 14C-labelled creatine uptake in isolated, incubated rat soleus (type I) muscle preparations at 37 degrees C. We found that the apparent Km for creatine uptake was 73 microM and the Vmax was 77 nmol h-1 gww-1. Creatine uptake was 82% inhibited by 2 mM beta-guanidinopropionic acid, the structural analogue of creatine. In addition, a decrease in buffer Na+ concentration, from 145 to 25 mM, reduced the rate of 14C-labelled creatine uptake by 77%, indicating that uptake is largely Na+-dependent in soleus muscle. Insulin had no effect on the rate of creatine uptake in vitro. The total creatine content was 34% lower, but the rate of creatine uptake in the presence of 100 microM extracellular creatine was 45% higher, in soleus than in extensor digitorum longus (type II) muscle. However, at 1 mM extracellular creatine, the maximal rate of uptake was not significantly different for the two muscle types, implying that soleus muscle has a lower Km for creatine uptake. We suggest that intracellular creatine levels may play a role in the regulation of skeletal muscle creatine uptake. PMID- 10383489 TI - Effects of anabolic/androgenic steroids on regenerating skeletal muscles in the rat. AB - We have examined the effect of male sexual hormones on the regeneration of skeletal muscles. Degeneration/regeneration of the left soleus and extensor digitorum longus muscles (EDL) of Wistar male rats was induced by an injection of snake venom (2 microg, Notechis scutatus scutatus). During the muscle regeneration (25 days), rats were treated with either oil (CON), nandrolone (NAN), NAN combined with exercise (NAN + EXE) or were castrated (CAS). Muscle growth and myosin heavy chain (MyHC) isoform content of regenerating muscles were studied. Castration altered the concentrations of MyHC in venom-treated EDL (P < 0.01) and soleus (P < 0.05). NAN increased the mass (P < 0.01) of regenerating soleus and decreased the relative amount of fast MyHC protein (% of total, P < 0.05). The effect of NAN + EXE on the fast MyHC proteins of venom-treated soleus was opposite (P < 0.05). NAN and NAN + EXE were without effect on the regenerating EDL (P > 0.05). In conclusion, it is possible that male sexual hormones play a role in the growth (synthesis of contractile proteins) of regenerating muscles in rat. In addition, contrary to NAN + EXE, NAN could be beneficial to soleus regeneration. PMID- 10383490 TI - Exogenous reactive oxygen and nitric oxide alter intracellular oxidant status of skeletal muscle fibres. AB - To test whether exogenous oxidants alter intracellular oxidant levels in skeletal muscle fibres, we exposed rat diaphragm to donors of nitric oxide (NOx), reactive oxygen species (ROS) or hyperoxia, and monitored intracellular oxidant levels using a fluorescent probe. Fibre bundles were dissected from the diaphragm and loaded with 2', 7'-dichlorodihydrofluorescein (DCFH); emissions were monitored using a fluorescence microscope. DCFH-loaded muscles were exposed to either a NOx donor (1 mM S-nitroso-N-acetyl penicillamine, SNAP; 1 mM sodium nitroprusside, SNP; 400 microM 1-hydroxy-2-oxo-3-(N-3-methyl-aminopropyl)-3-methyl-1-triazen, NOC-7), an ROS donor (100 microM hydrogen peroxide, H2O2; 100 microM tert-butyl hydroperoxide; 1 mM hypoxanthine plus 0.01 U mL-1 xanthine oxidase, HXXO) or a range of PO2s (25, 60 or 95% O2 oxygenating Krebs-Ringer solution) for 40 min; time-matched control bundles remained in Krebs-Ringer solution. Control muscles oxidized DCFH at a rate of 0.32 +/- 0.1 greyscale units min-1. SNAP (766%), SNP (1244%), NOC-7 (851%), H2O2 (543%), and HXXO (541%) increased DCFH oxidation from control levels. The increase in emissions caused by NOC-7 and SNP were blunted by the NOx scavenger haemoglobin (1 microM). DCFH oxidation by HXXO was unaffected by 1000 U mL-1 superoxide dismutase but was significantly decreased by 1000 U mL 1 catalase and 1 mM salicylate. PO2 had no effect on intracellular oxidant levels. Therefore, extracellular NOx and ROS can alter intracellular oxidant status in skeletal muscle fibres. These observations suggest that intrafibre oxidant levels could be the result of both intracellular and extracellular oxidant production. PMID- 10383491 TI - Near-infrared monitoring of tissue oxygenation during application of lower body pressure at rest and during dynamical exercise in humans. AB - During the application of a wide range of graded lower body pressures (LBP) (-50 to 50 mmHg), we examined how (1) the tissue oxygenation in the lower and upper parts of the body changes at rest, and (2) how tissue oxygenation changes in the lower extremities during dynamical leg exercise. We used near-infrared spectroscopy (NIRS) to measure the changes induced by LBP in total Hb content and Hb oxygenation in seven subjects. At rest, total Hb increased and Hb oxygenation decreased in the thigh muscles during -25 and -50 mmHg LBP, while both decreased during +25 and +50 mmHg LBP. However, in the forearm muscles during graded LBP, the pattern of change in total Hb was the reverse of that in the thigh. Measurements from the forehead showed changes only during +50 mmHg LBP. These results demonstrated that the pattern of change in total Hb and Hb oxygenation differed between upper and lower parts with graded LBP at rest. During dynamical leg exercise, total Hb and Hb oxygenation in the thigh muscles decreased during stepwise increases in LBP above -25 mmHg, Hb oxygenation decreasing markedly during +50 mmHg LBP. These results suggest that during dynamical exercise (i) LBP at +25 mmHg or more causes a graded decline in blood volume and/or flow in the thigh muscles, and (ii) especially at +50 mmHg LBP, the O2 content may decrease markedly in active muscles. Our results suggest that NIRS can be used to monitor in a non-invasive and continuous fashion the changes in oxygenation occurring in human skeletal muscles and head during the graded changes in blood flow and/or volume caused by changes in external pressure and secondary reflexes both at rest and during dynamical exercise. PMID- 10383492 TI - Involvement of the human splanchnic circulation in pressor response induced by handgrip contraction. AB - We have analysed the adjustment of blood flow and vascular conductance in the abundantly supplied splanchnic circulation to a generally released pressor reaction. Pressor responses were induced by 2-min periods of standardized, sustained handgrip in seven healthy students. The effects of handgrip tests were followed both in the fasting state and after the consumption of a substantial, mixed meal. In the first of the two sessions, changes in superior mesenteric artery blood flow were recorded and concomitant changes in local vascular conductance derived. In the other session, pressor released cardiac output changes were recorded and changes in total peripheral vascular conductance derived. Both types of flow changes were recorded using ultrasound Doppler technique. Typically, blood flow in the superior mesenteric artery increased two- to threefold after a meal. Handgrip contractions induced an initial rapid increase in heart rate, cardiac output and total peripheral conductance, followed by a gradual decline in total peripheral conductance and stroke volume and a gradual increase in heart rate and mean arterial pressure for the rest of the period. At the end of 2-min pressor periods, total peripheral conductance was only about 10% below the pre-handgrip level, whereas vascular conductance locally in the area of the superior mesenteric artery decreased by some 30%. Thus, it appears that the splanchnic vascular bed contributes markedly to the compound pressor response. Handgrips caused significantly less reduction in local vascular conductance in the post-prandial than in the pre-prandial state, indicating that blood flow to the digesting gastrointestinal tract retains a relatively high priority also in a pressor situation. PMID- 10383493 TI - Decrease in sodium-calcium exchange and calcium currents in diabetic rat ventricular myocytes. AB - This study was designed in order to gain insight into possible changes in the inward sodium-calcium exchange current (INa-Ca) and the L-type calcium current (ICa), in ventricular myocytes isolated from streptozotocin-induced diabetic rats. Recordings were made using the nystatin-perforated patch technique which minimizes interference with the normal intracellular Ca2+ buffering mechanisms. The averaged INa-Ca current density elicited by Ca2+ current was smaller in diabetic than in normal myocytes at all potentials tested. INa-Ca activated by rapid application of caffeine was significantly reduced and the decay phase was prolonged. The density of ICa was also significantly reduced by diabetes in the range of test potentials between -10 and +50 mV. In addition, the fast time constant of ICa inactivation, which represents mainly the sarcoplasmic reticulum (SR) Ca2+ release-induced inactivation, was significantly higher in diabetic than in normal myocytes. The decrease in ICa, which is the main source of trigger Ca2+ for SR Ca2+ release, may explain the significantly lowered peak systolic [Ca2+]i previously shown in diabetic myocytes. As activation of ICa is essential for subsequent stimulation of INa-Ca, reduced ICa may contribute to decreasing activation of the Na+-Ca2+ exchanger. PMID- 10383494 TI - Haemodynamic changes in the cardiovascular system during the early phases of orthostasis. AB - In this study, we monitored the changes in arterial blood pressure continuously in two groups of Caucasian men during normal passive orthostasis as well as reversed passive orthostasis. Group A consisted of a group of 23 younger men (16 +/- 0.5 years) and group B consisted of 21 older men (62.9 +/- 2.7 years). The normal passive orthostatic test and the reversed passive orthostatic test were used to induce blood pressure changes. We found that the temporary and initial changes in blood pressure during the normal and reversed orthostatic tests were significantly lower in the older group. Heart rate increases were also lower in the older group. These findings could be explained in terms of a reduced compliance of the thin walled venous blood vessels in the elderly. PMID- 10383495 TI - Hypoxia and hyperoxia both transiently affect distribution of pulmonary perfusion but not ventilation in awake sheep. AB - Despite a remarkable gravity independent heterogeneity in both local pulmonary ventilation and perfusion, the two are closely correlated at rest and during exercise in the normal lung. These observations strongly indicate that there is a mechanism for coupling of the two so that local V/Q-ratio is kept fairly uniform throughout the lung. This is also necessary to achieve adequate gas exchange in the lung. It was recently suggested that oxygen-induced vasoconstriction has a slow and intense component that might contribute to the matching of ventilation and perfusion also under normal conditions (Vejlstrup & Dorrington 1993). We therefore simultaneously determined distribution of local ( approximately 1(1/2) cm3 lung pieces) ventilation and perfusion in eight sheep at normoxia (FiO2 21%) and after 10 min and 2(1/2) h exposure to hypoxia (FiO2 12%; four sheep) or hyperoxia (FiO2 40%; four sheep). We used a approximately 1 microm wet fluorescent aerosol and 15 microm radioactive microspheres i.v. to measure local ventilation and perfusion, respectively. Neither hypoxia nor hyperoxia caused changes in the distribution of ventilation. After 10 min exposure to hypoxia or hyperoxia, distribution of perfusion was altered so that the correlation between values for local ventilation and perfusion decreased. After 2(1/2) h exposure to either hypoxia or hyperoxia, distributions of perfusion and V/Q-ratio had returned to baseline. These results show that distribution of perfusion is influenced by acute changes in oxygen tension, so that local matching of ventilation and perfusion is affected. Apparently, some mechanism restores the matching during extended exposure to the altered oxygen tension. PMID- 10383496 TI - Activation of renal pelvic chemoreceptors in rats: role of calcitonin gene related peptide receptors. AB - Substance P and calcitonin gene-related peptide (CGRP) increase afferent renal nerve activity (ARNA). A substance P receptor antagonist but not a CGRP receptor antagonist, h-CGRP (8-37), blocks the ARNA response to renal mechanoreceptor (MR) stimulation. We have examined whether calcitonin gene-related peptide activates renal pelvic sensory receptors and whether such activation contributes to renal chemoreceptor stimulation. The calcitonin gene-related peptide receptor antagonist, h-CGRP (8-37) [0.01-10 micromol L-1] dose-dependently decreased (29 +/- 4-86 +/- 13%, P < 0.01) the ipsilateral afferent renal nerve activity in response to the renal pelvic administration of calcitonin gene-related peptide (0.26 micromol L-1). Renal pelvic perfusion with 900 mM NaCl also increased ipsilateral ARNA (23 +/- 3% increase, P < 0.02) and contralateral urinary sodium excretion (13 +/- 4% increase, P < 0. 05). However, these responses to hypertonic NaCl were unaltered by h-CGRP (8-37). Renal pelvic perfusion with 1 or 10 microM h-CGRP (8-37) also failed to alter the ARNA responses to KCl (31.25, 62.5 and 125 mM). These results indicate that there are sensory receptors in the renal pelvic area that are responsive to calcitonin gene-related peptide. The activation of these receptors elicits a contralateral natriuretic response. In contrast, the activation of renal calcitonin gene-related peptide receptors does not contribute to renal chemoreceptor activation. PMID- 10383497 TI - Personal review: Helicobacter pylori, NSAIDs and cognitive dissonance. AB - Helicobacter pylori and non-steroidal anti-inflammatory drugs (NSAIDs) each cause peptic ulcers but by different mechanisms. As a result, the effect of both of these risk factors together is not a synergistic enhancement of injury, ulceration or rates of complications. Indeed, there are circumstances under which patients infected with H. pylori are less prone to NSAID-induced ulcers than those who are not infected or who have undergone eradication treatment. This may be because of opposite effects on gastric mucosal prostaglandin synthesis or for other reasons. Reluctance to accept that there may be specific circumstances where H. pylori is beneficial may arise because of the psychological process of cognitive dissonance. PMID- 10383498 TI - The GU-MACH study: the effect of 1-week omeprazole triple therapy on Helicobacter pylori infection in patients with gastric ulcer. AB - AIMS: To study the efficacy of omeprazole triple therapy in the eradication of Helicobacter pylori in patients with active gastric ulcer, and to assess healing and relapse of gastric ulcer. METHODS: A double-blind, randomized study was carried out in 18 centres in Germany, Hungary and Poland. Patients (n = 160) with gastric ulcer and a positive H. pylori screening test were randomized to a 7-day twice daily treatment with omeprazole 20 mg, clarithromycin 500 mg and amoxycillin 1000 mg (OAC) or omeprazole 20 mg, clarithromycin 250 mg and metronidazole 400 mg (OMC), or with omeprazole 20 mg once daily (O). After completion of this 1-week treatment, patients were treated with omeprazole until healing (maximum 12 weeks), and followed for 6 months. H. pylori was assessed by urea breath test (UBT) and histology. RESULTS: Eradication rates ITT were OAC 79% (95% CI: 65-90%), OMC 86% (95% CI: 73-94%) and O 4% (95% CI: 0-14%). Eradication rates PP were OAC 83% (95% CI: 68-93%), OMC 93% (95% CI: 80-98%) and O 3% (95% CI: 0-13%). Gastric ulcer relapses occurred in 5, 0 and 11 patients in the groups, respectively. CONCLUSIONS: The results from the study demonstrate that OMC and OAC 1-week regimens are safe and effective for eradication of H. pylori in gastric ulcer patients, and that ulcer relapse is infrequent after successful eradication. PMID- 10383499 TI - Antibiotic combination therapy in patients with chronic, treatment-resistant pouchitis. AB - BACKGROUND: Pouchitis is the major long-term complication after ileal pouch-anal anastomosis for ulcerative colitis. About 15% of patients have a chronic, treatment-resistant disease. AIMS: To evaluate the efficacy of an antibiotic combination for chronic active, treatment-resistant pouchitis. PATIENTS AND METHODS: Eighteen patients were treated orally with rifaximin 1 g b.d. + ciprofloxacin 500 mg b.d. for 15 days. Symptoms assessment, endoscopic and histological evaluations were performed at screening and after 15 days using the Pouchitis Disease Activity Index (PDAI). Improvement was defined as a decrease of at least 3 points in PDAI score, and remission as a PDAI score of 0. Systemic absorption of rifaximin was determined by high performance liquid chromatography. Faecal samples were collected before and after antibiotic treatment for stool culture. RESULTS: Sixteen out of 18 patients (88.8%) either improved (n=10) or went into remission (n=6); the median PDAI scores before and after therapy were 11 (range 9-17) and 4 (range 0-16), respectively (P < 0.002). No side-effects were reported. Rifaximin plasma levels and urinary excretion were negligible, confirming its mainly topical activity. A significant decrease in total anaerobes and aerobes, enterococci, lactobacilli, bifidobacteria and bacteroides in faecal samples was observed, while the reduction in number of coliforms and Clostridium perfringens did not reach a statistical significance. CONCLUSIONS: A combination of rifaximin and ciprofloxacin was effective in patients with active chronic, treatment-resistant pouchitis, suggesting the need, in these patients, for treatment using antibiotic agents with wide antibacterial spectrum of activity. PMID- 10383500 TI - The importance of clarithromycin dose in the management of Helicobacter pylori infection: a meta-analysis of triple therapies with a proton pump inhibitor, clarithromycin and amoxycillin or metronidazole. AB - BACKGROUND: It is not clear which dose of clarithromycin (500 mg b.d. or 250 mg b.d.) is more effective for Helicobacter pylori eradication in proton pump inhibitor-based triple therapies. METHODS: We undertook a meta-analysis of the effect of 7-day triple therapies consisting of a proton pump inhibitor (P), and clarithromycin (C) and amoxycillin (A) or metronidazole (M). A meta-analysis of all clinical trials performed in an adult population and published in English up to March 1998 was undertaken. Studies with doses of clarithromycin 500 mg b.d. or 250 mg b.d. only were included. RESULTS: A total of 82 studies (31 papers and 51 abstracts) involving 110 treatment arms and 6123 patients were analysed that met the predetermined inclusion and exclusion criteria. In the PAC combination, the pooled eradication rate in patients treated with clarithromycin 500 mg b.d. was 89.5% (95% CI: 86.9-92. 0%) by per protocol analysis and 86.6% (95% CI: 81.0 89.3%) by intention-to-treat analysis. These rates are significantly higher than those achieved with clarithromycin 250 mg b.d. (83.3% by per protocol and 78.2% by intention-to-treat analysis, both P < 0.0001). This difference was confirmed in head-to-head comparative studies. In the PMC regimen, clarithromycin 500 mg b.d. eradicated 90.8% (95% CI: 87.0-94.5%) of the infections compared to 88.5% (95% CI: 85.5-91. 5%) in patients treated with clarithromycin 250 mg b.d. by per protocol analysis (P = 0.082). The corresponding rates by intention-to-treat analysis for clarithromycin 500 mg b.d. and 250 mg b.d. was 88.3% and 86.7%, respectively (P = 0.259). CONCLUSIONS: Seven-day triple therapies with a proton pump inhibitor, clarithromycin and amoxycillin or metronidazole are highly effective treatments for the eradication of H. pylori. Clarithromycin 500 mg b. d. should be used in these combinations to achieve the best first treatment results, which can minimize the subsequent development of bacterial resistance to clarithromycin and metronidazole. PMID- 10383501 TI - The effect of Helicobacter pylori eradication on intragastric pH during dosing with lansoprazole or ranitidine. AB - BACKGROUND: The antisecretory effect of omeprazole on intragastric pH is decreased in the absence of Helicobacter pylori. AIM: To investigate the effect of H. pylori eradication on intragastric pH during lansoprazole or ranitidine dosing in 41 asymptomatic H. pylori-positive subjects. METHOD: Two groups of healthy H. pylori-positive volunteers were investigated. One group was dosed with lansoprazole 30 mg at 08.00 hours for at least 8 days, before and after 2 weeks of placebo-controlled double-blind eradication therapy using ranitidine bismuth citrate 400 mg b.d. and clarithromycin 500 mg b.d. The other group was dosed with ranitidine 300 mg at 23.00 hours for at least 8 days using the same trial design. An upper endoscopy was performed to establish H. pylori status by rapid urease test, culture and histology before both periods of dosing. Twenty-four hour intragastric pH recording was performed on the final day of all periods of dosing. RESULTS: H. pylori eradication significantly decreased the intragastric pH reached during lansoprazole treatment throughout all periods of the day. Intragastric pH during ranitidine treatment was not affected by H. pylori eradication, except for the late-night period. CONCLUSION: H. pylori eradication has a more pronounced effect on the acid-inhibiting properties of lansoprazole than on those of ranitidine. PMID- 10383502 TI - Impact of rabeprazole, a new proton pump inhibitor, in triple therapy for Helicobacter pylori infection-comparison with omeprazole and lansoprazole. AB - BACKGROUND: A recent trend in curative therapy for Helicobacter pylori infection is the so-called triple therapy, which consists of a proton pump inhibitor (PPI) and two different antimicrobials. Various regimens employing this triple therapy have been reported. However, little is known about the effectiveness of rabeprazole, a recently developed proton pump inhibitor, when used in the triple therapy. AIM: To validate its usefulness by comparing rabeprazole with omeprazole and lansoprazole, in combination with amoxycillin and clarithromycin. PATIENTS AND METHODS: 221 H. pylori-positive patients with peptic ulcer disease were randomized to receive one of three different proton pump inhibitor/amoxycillin clarithromycin (PPI/AC) regimens for 7 days. (i) OAC regimen (n = 75): omeprazole 20 mg b.d., amoxycillin (AMOX) 500 mg t.d.s. and clarithromycin (CAM) 200 mg b.d.; (ii) LAC regimen (n = 74): lansoprazole 30 mg b.d. , AMOX 500 mg t.d.s. and CAM 200 mg b.d.; and (iii) RAC regimen (n = 72): rabeprazole 20 mg b.d., AMOX 500 mg t.d.s. and CAM 200 mg b.d. Cure of the infection was determined by the 13C urea breath test 1 month after completion of the treatment. RESULTS: Intention-to treat based cure rates for OAC, LAC and RAC regimens were 85% (95% CI, 75-92), 84% (95%, CI 73-91) and 88% (95% CI, 78-94), respectively, and per protocol based cure rates of these regimens were 88% (95% CI, 78-94), 91% (95%, CI 82-99) and 93% (95% CI, 84-98), respectively. Adverse effects in the entire study population, which included diarrhoea, glossitis or skin rash, were reported by 15% of the patients, and complete compliance was achieved in 95% of these patients. CONCLUSION: 1-week proton pump inhibitor/AC regimens for H. pylori infection were effective in the Japanese population. Rabeprazole can be considered as equivalent to omeprazole and lansoprazole in the PPI/AC triple therapy. PMID- 10383503 TI - A multicentre evaluation of the laser assisted ratio analyser (LARA): a novel device for measurement of 13CO2 in the 13C-urea breath test for the detection of Helicobacter pylori infection. AB - BACKGROUND: The laser assisted ratio analyser (LARA) was developed as a novel device to measure 13CO2 in the urea breath test for the detection of H. pylori infection. The analyser was tested in a prospective multicentre study in 444 patients in North America (Phase 1) followed by second study involving 160 patients (Phase 2). METHODS: Patients undergoing endoscopy for clinical indications had antral and gastric biopsies taken for histological examination, culture and CLO test. One hour after endoscopy, a baseline breath sample was obtained, 100 mg of 13C-urea were ingested and breath samples were obtained at 30 and 60 min post ingestion. Data obtained with the LARA were compared with the results of culture, rapid urease testing and central pathology in two different combinations {reference standards}. The study was conducted in two phases: in Phase 2, a modification was made to the LARA that improved the removal of water vapour from the breath sample. RESULTS: In Phase I, data from 331 patients were analysed using a cut off of (delta) 7.8 +/- 0.8, the sensitivity of the method was 91.7% and the specificity was 86.5%, using the reference standard of 2 of 3 tests (CLO, culture or histology) being positive. Positive and negative predictive values were, respectively, 85.2% and 92.5%. In Phase 2 of the study, 160 patients were enrolled and 141 patients were analysed using the same standards. We used the same reference standards but with a cut off of (delta) 6.1 +/- 0.6. The sensitivity and specificity increased to 96.8% and 98.6%, respectively. Positive and negative predictive values were, respectively, 98.4% and 97.3%. The detection rates for H. pylori were similar in patients with peptic ulcer or H. pylori associated gastritis. CONCLUSIONS: The LARA provides an accurate non-invasive means of detecting 13CO2 in the 13C-urea breath test for H. pylori in a multicentre clinical environment that compares well with invasive 'gold standard' methods. PMID- 10383504 TI - Effects of ranitidine bismuth citrate on Helicobacter pylori motility, morphology and survival. AB - AIM: The effects of the anti-ulcer agents ranitidine bismuth citrate (RBC), ranitidine hydrochloride (R) and colloidal bismuth citrate (BC), on Helicobacter pylori motility, morphology and survival were examined to determine whether the clinical effectiveness of RBC might be linked to a specific action that inhibits bacterial motility. METHODS: H. pylori from patients with duodenal ulcer or non ulcer dyspepsia were exposed to RBC and BC at bismuth concentrations ranging from 12.5 to 50 microg/mL, and R at ranitidine concentrations ranging from 12.5 to 50 microg/mL for a brief period (< 15 min), 6 h and 24 h. Bacterial motility was assessed with a Hobson BacTracker, bacterial morphology by transmission electron microscopy, and growth inhibition by counting colony-forming units. RESULTS: H. pylori motility was diminished with RBC and BC but not R. However, the effect of RBC was markedly greater than that of BC at each bismuth concentration and time of exposure tested: (i) brief exposure to RBC/bismuth 50 microg/mL but not to BC, resulted in a significant loss of motility without loss of viability or change in cell morphology, and (ii) bacteria were immobilized, and lost viability after exposure to RBC/bismuth 50 microg/mL for 24 h but not to BC. Morphological destruction caused by RBC differed from that by BC: after 24 h exposure to the highest concentration tested, cell fragmentation and flagella detachment occurred more frequently with BC than RBC, but the latter produced greater disruption of intracellular structures. CONCLUSIONS: RBC suppresses growth of H. pylori, and has a specific inhibitory effect on the bacterial motor mechanism. These pharmacological actions are likely to contribute to the clinical effectiveness of the agent. PMID- 10383505 TI - Specific inhibition of cyclooxygenase-2 with MK-0966 is associated with less gastroduodenal damage than either aspirin or ibuprofen. AB - BACKGROUND: Compared with currently available NSAIDs (which inhibit COX-1 and COX 2 isoforms of cyclooxygenase), MK-0966 (a specific COX-2 inhibitor) is expected to cause less gastrointestinal toxicity. AIM: To compare the effect on the upper gastrointestinal mucosae of a high dose of MK-0966 with that of conventional doses of ibuprofen and aspirin. METHODS: Healthy subjects (n = 170; age range 18 54 years) with endoscopically normal gastric and duodenal mucosa were randomized to either MK-0966 250 mg q.d. (n = 51), ibuprofen 800 mg t.d.s. (n = 51), aspirin 650 mg q.d.s. (n = 17), or placebo (n = 51) in this 7-day, double-blind, parallel group study. The mucosae were evaluated by endoscopy using a predefined scale; scores could range from 0 to 4. The primary end-point was the percentage of subjects who developed a mucosal score >/= 2 (i.e. the development of one or more erosions). To evaluate COX-1 activity, serum thromboxane B2 levels were determined in a subset of the population. RESULTS: The percentage of subjects who developed a mucosal score >/= 2 in the MK-0966 group (12%) was significantly lower (P < 0.001) than that in the ibuprofen (71%) and aspirin (94%) groups, and was similar to that in the placebo group (8%). Only ibuprofen and aspirin significantly (P < 0.0001) reduced baseline thromboxane B2 levels. All treatments were generally well tolerated. CONCLUSIONS: In this acute short-term endoscopic study, MK-0966 250 mg q.d. (a dose at least 10 times higher than that demonstrated to reduce the signs and symptoms of osteoarthritis) produced significantly less gastrointestinal mucosal damage than either ibuprofen 800 mg t.d.s. or aspirin 650 mg q.d.s. and was comparable to placebo in this regard. PMID- 10383506 TI - A new quadruple therapy for Helicobacter pylori: influence of resistant strains on treatment outcome. AB - BACKGROUND: There have been no reports concerning the efficacy and safety of a 1 week quadruple therapy regimen of omeprazole, amoxycillin, roxithromycin and metronidazole for Helicobacter pylori infections and the impact of primary resistance on the eradication rate. METHODS: One hundred and sixty-nine consecutive patients with peptic ulcer disease as well as gastritis with biopsy proven H. pylori infection were entered into an open study of omeprazole 20 mg o.m., amoxycillin 500 mg t.d.s., roxithromycin 150 mg b.d., and metronidazole 250 mg t.d.s. Helicobacter pylori status was determined by urease test, histology and culture. Susceptibility to amoxycillin, metronidazole and roxithromycin was determined by the E-test. RESULTS: H. pylori was eradicated in 155 out of 169 (92%; 95% CI 88-96%) by intention-to-treat analysis, and in 155 out of 163 (95%; 95% CI 92-98%) by per protocol analysis. The prevalence of primary resistance against amoxycillin, roxithromycin and metronidazole was 2 out of 166 (1%), 16 out of 166 (10%) and 27 out of 166 (16%), respectively. H. pylori was eradicated in 25 out of 27 (93%) patients with metronidazole-resistant strains compared with 130 out of 136 (96%) in patients with metronidazole-sensitive strains of H. pylori. It was eradicated in 15 out of 16 (94%) patients with roxithromycin resistant strains while in 140 out of 147 (95%) patients with roxithromycin sensitive strains of H. pylori, and in two out of two (100%) patients with amoxycillin-resistant stains compared with 153 out of 161 (95%) in patients with amoxycillin-sensitive strains. H. pylori was eradicated in three out of four (75%) patients with double resistance against metronidazole and roxithromycin compared with 152 out of 159 (96%) patients with sensitive strains to metronidazole and or roxithromycin. None of these differences were statistically significant. Severe side-effects were found in only one out of 169 patients anaphylaxis due to penicillin. CONCLUSIONS: The 1-week quadruple therapy with omeprazole, amoxycillin, metronidazole and roxithromycin was found to eradicate H. pylori in over 90% of all patients. This regimen was also found to be beneficial for patients with pre-treatment resistant strains to metronidazole, roxithromycin or amoxycillin, and was observed to be safe and well-tolerated. PMID- 10383507 TI - Impact of clarithromycin resistance on the effectiveness of a regimen for Helicobacter pylori: a prospective study of 1-week lansoprazole, amoxycillin and clarithromycin in active peptic ulcer. AB - BACKGROUND: Clarithromycin is a key antimicrobial in the combinations used to cure Helicobacter pylori infections, so there is a need to define the impact of in vitro resistance on in vivo results. METHODS: A prospective trial was designed to study the effectiveness of the 1-week combination of lansoprazole, clarithromycin and amoxycillin in 102 consecutive patients with active peptic ulcer. The pre-treatment and post-treatment sensitivity to amoxycillin, metronidazole and clarithromycin were studied by E-test, and H. pylori status was defined by histology, culture and urease test at diagnosis and one month after treatment, and by urea-breath test 2 months after treatment. RESULTS: The eradication rate (intention-to-treat analysis) was 77% (95% CI: 69-86). No clinical factor was found to be different between eradicated and non-eradicated patients. Clarithromycin-resistant strains were found in 10 (10%; CI: 5-17) patients. The eradication rate was 20% (CI: 3-56) in these patients vs. 83% (CI: 75-91) in patients harbouring clarithromycin-sensitive strains (P < 0.001). A logistic-regression analysis confirmed clarithromycin resistance as the only factor associated with treatment failure. CONCLUSIONS: Clarithromycin resistance significatively impairs the effectiveness of the combination of lansoprazole, amoxycillin, and clarithromycin. The 80% efficacy goal will be difficult to reach in areas with high (>10%) primary clarithromyicin resistance, if currently recommended proton pump inhibitor-triple therapies are used. PMID- 10383508 TI - Two-week dual vs. one-week triple therapy for cure of Helicobacter pylori infection in primary care: a multicentre, randomized trial. AB - BACKGROUND: One-week triple therapy has been suggested to be superior to two-week omeprazole-clarithromycin therapy for the cure of Helicobacter pylori infection. However, direct comparisons of the two treatments are scarce. AIM: To compare triple with dual therapy for H. pylori infection in the primary care setting. METHODS: One hundred and forty-five patients with duodenal ulcer and H. pylori infection were randomized to receive omeprazole 20 mg b.d. and clarithromycin 500 mg t.d.s. for 14 days (OC14 group, 69 patients) or omeprazole 20 mg b.d., clarithromycin 500 mg b.d. and amoxycillin 1 g b.d. for 7 days (OCA7 group, 76 patients). Eradication was evaluated by the 13C-urea breath-test. RESULTS: Intention-to-treat analysis showed a cure rate of 48% (95% CI: 36-60%) in the OC14 group, and 71% (95% CI: 59-80%) in the OCA7 group (P=0.0004). Per protocol analysis showed cure rates of 51% (95% CI: 38-63%, 33/65 patients) and 82% (95% CI: 70-90%, 54/66 patients), respectively (P=0.0001). There were no significant differences in compliance or side-effects. CONCLUSION: One-week twice-daily triple therapy is superior to 2-week dual therapy, but the cure rate in primary care was far below 90%. PMID- 10383509 TI - Cisapride 20 mg b.d. for preventing symptoms of GERD induced by a provocative meal. The CIS-USA-89 Study Group. AB - BACKGROUND: Cisapride is a substituted piperidinyl benzamide indicated for the symptomatic treatment of patients with nocturnal heartburn due to gastro oesophageal reflux disease (GERD). The currently recommended dosing regimen for cisapride is 10 mg q.d.s., but the elimination half-life of 8-10 h supports b.d. dosing with 20 mg. METHODS: This multicentre, randomized, double-blind, placebo controlled trial was undertaken to determine the efficacy and safety of cisapride 20 mg b.d. dosing in reducing or preventing heartburn and other meal-related symptoms after challenge with a provocative fatty meal. In phase 1 of the study, 137 patients with at least a 3-month history of symptoms suggestive of GERD and at least five episodes of GERD on 7-day diary were eligible to receive single blind treatment with placebo for 7 (range +/- 3) days and then ingested a provocative meal. One hundred and twenty-two patients (45 men and 77 women, 22-65 years of age) who experienced heartburn during the 3 h after ingestion of the meal qualified for the double-blind phase of the study and were randomly assigned to either cisapride 20 mg or matching placebo b.d. for 7 (+/-3) days. At the end of this period, 118 patients again ate a fatty meal and were assessed for symptoms of GERD. RESULTS: Heartburn was prevented in a significantly higher percentage of cisapride-treated patients (40%; 24 out of 60) than placebo-treated patients (21%; 12 out of 58) after the repeat provocative meal at the end of the double-blind phase (P = 0.017). Cisapride was also significantly more effective in reducing the severity of postprandial heartburn, belching, and regurgitation (P < 0.05). Twice-daily dosing with cisapride 20 mg was well tolerated; the number of cisapride- and placebo-treated patients who experienced at least one adverse event was similar (31% and 22%, respectively). The most common adverse events were diarrhoea (cisapride, 18%; placebo, 0%) and rhinitis (cisapride, 2%; placebo, 5%). CONCLUSIONS: These results demonstrate that cisapride 20 mg b.d. is effective in preventing or reducing symptoms of heartburn in patients who developed heartburn after ingesting a provocative fatty meal. Cisapride was also effective in reducing the severity of heartburn-related symptoms such as belching and regurgitation. PMID- 10383510 TI - Lansoprazole in the treatment of heartburn in patients without erosive oesophagitis. AB - BACKGROUND: This randomized, double-blind, multicentre study compared lansoprazole with placebo for symptomatic relief of patients with non-erosive gastro-oesophageal reflux disease (GERD). METHODS: 214 patients with symptomatic, non-erosive GERD (moderate to severe daytime and/or night-time heartburn greater than half the days over the past 6 months and during the 7- to 10-day pre treatment period) were randomized to either lansoprazole 15 mg or lansoprazole 30 mg, or placebo o.d. for 8 weeks. RESULTS: Daily diary data indicated that on the first treatment day a statistically significantly smaller percentage of lansoprazole patients reported daytime and night-time heartburn and antacid usage, compared with placebo patients. Lansoprazole patients also reported statistically significant less severe daytime and night-time heartburn on the first treatment day. During 0-4, 4-8, and 0-8 weeks of therapy, a statistically significant smaller percentage of days and nights with heartburn, less severe daytime and night-time heartburn, and less antacid usage were observed in the lansoprazole group compared to the placebo group. The percentages of patients with adverse reactions were similar in the lansoprazole and placebo groups. CONCLUSIONS: The results of this study demonstrate that lansoprazole is an appropriate therapy for patients with symptomatic non-erosive GERD. PMID- 10383512 TI - Survey on repeat prescribing for acid suppression drugs in primary care in Cornwall and the Isles of Scilly. AB - BACKGROUND: Repeat prescriptions for acid suppression therapy represent an important burden on health care resources. AIM: To determine the prevalence of acid suppression therapy and its indications by general practitioners (GPs) in a larger sample of practices than previous studies. METHOD: Practices in Cornwall and the Isles of Scilly were invited to identify the number of patients on repeat prescription for acid suppression drugs in their practice, to review the indication for treatment in a sample of 50 patients, and to indicate the mode of review of these patients. RESULTS: Out of 77 practices, 42 (55%) participated in the study. Overall, 5% of patients were authorized to receive a repeat prescription for acid suppression drugs. Repeat rates varied between practices, from 1.68% to 11.11%. Repeat rates increased with age and were higher in men than in women. Only 41% of patients had a proven diagnosis of gastro-oesophageal reflux disease or peptic ulcer. A review of notes was the most frequent way (36%) stated by GPs to review acid suppression therapy. CONCLUSION: The repeat rate found in our study was higher than that found in previous studies. A high proportion of older patients in Cornwall, as well as a continuing increase in the prescription of acid suppression drugs, may account for these results. PMID- 10383511 TI - Lack of effect of spearmint on lower oesophageal sphincter function and acid reflux in healthy volunteers. AB - BACKGROUND: Spearmint is commonly used as an antispasmodic and as a flavouring in several medications including antacids. It can produce heartburn, presumably by lowering lower oesophageal sphincter (LES) tone, but the mechanism has not previously been objectively examined. AIM: To study the effect of spearmint on LES function, acid reflux and symptoms. METHODS: In healthy volunteers, a Dent Sleeve and a pH electrode were placed in the distal oesophagus. They were then given spearmint either in a flavouring (0.5 mg), or a high (500 mg) dose, or a placebo, using a double-blind randomized crossover design. LES pressure, oesophageal pH and symptoms were recorded for 30 min before and after administration. RESULTS: LES pressure was not affected by spearmint, either high dose (19.6 vs. 16.0 mmHg), flavouring dose (20.2 vs. 19.8 mmHg) or placebo (20.5 vs. 19.2 mmHg, all N.S.). There were no differences in reflux occurrence following high dose (mean = 0.65 vs. 0.85 episodes), low dose (0.4 vs. 0.5 episodes) or placebo (0.7 vs. 1.10 episodes, all N.S.). There was a significant increase in mean symptom scores following high-dose spearmint (0 vs. 0.35, P = 0.03), but not low dose (0 vs. 0.2) or placebo (0 vs. 0.5, both N.S.). One subject reported symptoms with placebo, one with low dose, and six with high dose; all without increased reflux episodes or decreased sphincter pressure. CONCLUSION: Spearmint has no effect on LES pressure or acid reflux. Flavouring doses of spearmint do not produce more symptoms than placebo while high doses can be associated with symptoms, presumably from direct mucosal irritation but not reflux. PMID- 10383513 TI - Efficacy of omeprazole versus ranitidine for symptomatic treatment of poorly responsive acid reflux disease-a prospective, controlled trial. AB - BACKGROUND: H2-receptor antagonists are widely used in patients with gastro oesophageal reflux disease (GERD) and are frequently continued when symptoms persist. AIM: To compare the efficacy of omeprazole 20 mg once daily with that of ranitidine 150 mg twice daily in relieving GERD symptoms, in patients who remained symptomatic following a 6-week course of ranitidine therapy. METHODS: Patients with heartburn on at least 4 days/week but who did not have endoscopy to assess oesophageal mucosa could participate. This two-phase, prospective trial included a 6-week open-label phase (phase I), followed by an 8-week double-blind phase (phase II). Patients still symptomatic following treatment with ranitidine 150 mg twice daily (phase I) were randomized to double-blind treatment (phase II) with either omeprazole 20 mg once daily or ranitidine 150 mg twice daily. The primary efficacy variable was the proportion of patients with heartburn resolution during weeks 4 and 8 of phase II. RESULTS: Of the 533 patients with GERD who received ranitidine in phase I, 348 patients (65%) were still symptomatic. A total of 317 patients (59%) were randomized to double-blind treatment (phase II). At week 8, a significantly (P < 0.0004) greater proportion of omeprazole-treated patients (70%) experienced no more than mild heartburn compared with ranitidine-treated patients (49%). Complete resolution of heartburn also occurred in a significantly (P < 0. 00001) greater proportion of omeprazole treated patients (46% vs. 16% of the ranitidine group at week 8). CONCLUSIONS: After 6 weeks of ranitidine treatment, the majority of patients with GERD were still experiencing moderate to severe heartburn. Omeprazole was significantly more effective than ranitidine in resolving heartburn in this group of patients. PMID- 10383514 TI - Lansoprazole is superior to ranitidine as maintenance treatment for the prevention of duodenal ulcer relapse. AB - AIM: To compare lansoprazole 30 mg once daily, lansoprazole 15 mg once daily and ranitidine 150 mg once nightly in the prevention of duodenal ulcer relapse in patients whose duodenal ulcers had been previously healed with lansoprazole 30 mg once daily or ranitidine 300 mg nightly. METHODS: A double-blind, parallel group, randomized multicentre study conducted in 33 centres in the UK, Eire, Sweden and Australia. Two hundred and nineteen patients with a duodenal ulcer were randomized to receive lansoprazole 30 mg and 217 to receive ranitidine 300 mg for 8 weeks. Patients were then re-randomized to receive lansoprazole 30 mg (122 patients), lansoprazole 15 mg (121 patients) or ranitidine 150 mg (116 patients) for 12 months. All patients had an endoscopically-proven duodenal ulcer at baseline and were considered suitable for long-term maintenance therapy to prevent relapse. RESULTS: Significantly more patients were healed on lansoprazole (98%) compared to ranitidine (89%) (P < 0.001, Fisher's exact test). Lansoprazole provided more rapid symptom relief than ranitidine. Lansoprazole 30 mg and lansoprazole 15 mg increased the probability of not relapsing in comparison to ranitidine (P = 0.001 and 0.06, respectively, life-table analysis). Relapse rates over the 12 months were lower in the lansoprazole treatment groups (lansoprazole 30 mg, 5%; lansoprazole 15 mg, 12%; and ranitidine, 21%; lansoprazole 30 mg vs. ranitidine 150 mg, P = 0.002). Symptoms were well controlled in both groups during the maintenance phase. All treatments were well tolerated with no major differences seen in adverse event profiles between treatment groups. CONCLUSIONS: Both doses of lansoprazole (30 mg and 15 mg) were superior to ranitidine 150 mg in the prevention of duodenal ulcer relapse. Lansoprazole was superior to ranitidine in terms of symptom control and duodenal ulcer healing. Both treatments were well tolerated. PMID- 10383515 TI - Changes in gastric mucosal ulcerogenic responses in rats with adjuvant arthritis: role of nitric oxide. AB - AIM: To examine gastric mucosal ulcerogenic responses to indomethacin and HCl/ethanol in adjuvant arthritic (AA) rats. METHODS AND RESULTS: Arthritis was induced in male Dark Agouti (DA) rats by injection of Freund's complete adjuvant (FCA) into the right hind paw. The gastric ulcerogenic response to indomethacin was markedly worsened in AA rats, depending on the degree of arthritic change. This aggravation of indomethacin-induced gastric lesions in AA rats was significantly prevented by NG-nitro-L-arginine methyl ester (L-NAME) and amino guanidine as well as dexamethasone. In contrast, the mucosal ulcerogenic response to HCl/ethanol was inhibited in AA rats. The suppression of HCl/ethanol-induced gastric lesions in AA rats was reversed almost totally by L-NAME and aminoguanidine as well as dexamethasone and partly by indomethacin. The expression of inducible nitric oxide synthase (iNOS) mRNA was observed in the stomach of AA rats but not of normal rats. Moreover, the luminal releases of nitric oxide (NO) metabolites as well as prostaglandin (PG) E2 were significantly increased in AA rats. CONCLUSIONS: The gastric mucosal ulcerogenic responses were modified in AA rats, in different manners depending on the irritants; an increase in response to indomethacin and a decrease in response to HCl/ethanol. These changes may both be accounted for by increased production of NO by iNOS, and the latter is also partly related to increased production of PGs. PMID- 10383516 TI - Review article: the management of Giardiasis. AB - Giardiasis is the intestinal infection resulting from infestation with the human parasite Giardia intestinalis, also called Giardia lamblia. The infection may be asymptomatic or present with a variety of symptoms such as diarrhoea, weight loss, abdominal cramps, and failure to thrive. Giardiasis is most often diagnosed after recent travel or in day care centres. The organism has two stages in its life cycle. It is usually ingested as a cyst with as few as 10-25 cysts being sufficient to cause infection. After excystation, the organism is a replicative trophozoite which may attach to the small bowel wall. Giardia intestinalis does not invade the bowel wall. Trophozoites may encyst and be shed in faeces for future ingestion by another host. Diagnosis of infection is by stool examination which may also eliminate other possible infectious agents. Small bowel biopsy may be necessary in difficult individual cases or to rule out non-infectious illnesses, and stool ELISA may serve for large population screening examinations. The mainstay of treatment is metronidazole 250-400 mg three times per day by mouth for 5 days. PMID- 10383517 TI - Helicobacter pylori infection and gastric cancer: systematic review of the epidemiological studies. AB - BACKGROUND: The published epidemiological studies of chronic Helicobacter pylori infection and gastric cancer yield conflicting results, so there is uncertainty as to whether any material association exists and, if so, how strong it is. AIM: To review these studies quantitatively. METHODS: A systematic review of sero epidemiological studies published before 1998 of H. pylori and gastric cancer, as identified by computer-assisted literature searches of relevant journals, reference lists and discussions with authors. All relevant studies identified were included, subdivided by study design. The following was abstracted from published reports: adjusted odds ratio (or, in prospective studies, the risk ratio) and confidence interval, study design, type of controls, mean age, mean duration of follow-up, assay methods, location of study, and degree of adjustment for confounders. RESULTS: The 34 retrospective studies included in total 3300 gastric cancers, but their controls were of uncertain validity. The 10 'nested' case-control comparisons in prospective studies included in total only 800 gastric cancers, and combined analysis of them yielded a risk ratio of 2.5 (95% CI: 1.9-3.4; 2P < 0.00001) for gastric cancer in people seropositive for H. pylori antibodies. CONCLUSIONS: The prospective studies suggest that gastric cancer is 2 or 3 times as common in those chronically infected by H. pylori, but to help investigate causality, further observational studies are still needed, as are large-scale randomized trials of whether antibacterial regimens reduce the eventual incidence of gastric cancer. PMID- 10383518 TI - Evaluation of treatment regimens to cure Helicobacter pylori infection--a meta analysis. AB - OBJECTIVE: To assess effectiveness of treatment to cure Helicobacter pylori infection. DATA SYNTHESIS: Meta-analysis of 666 manuscripts (full papers, abstracts, letters to the editor) identified through Medline and a manual search (1986 to January 1998). Data were overviewed by regression analysis with weighted random effects models. SUBJECTS: 53 228 patients with H. pylori infection. INTERVENTIONS: Patients were treated with 132 different medication combinations. MAIN OUTCOME MEASURE: Cure of H. pylori infection per protocol and intention-to treat basis at least 28 days after treatment. RESULTS: The nationality of the patients and therapeutic regimen have a significant impact on the results, after correction for the heterogeneity in the precision of the cure rate caused by different study sizes and random effect for study. On the basis of the original sample size, cure rates of 80-85% were achieved using combinations of a proton pump inhibitor or ranitidine bismuth citrate with two antibiotics including clarithromycin, amoxycillin and metronidazole or tinidazole. Comparable cure rates were also achieved using a combination of a proton-pump inhibitor or H2 receptor antagonist with bismuth subcitrate or tripotassium dicitrato bismuthate, metronidazole and tetracycline. The dose of clarithromycin influenced cure rates. Treatment duration did not influence the outcome. CONCLUSION: Several therapeutic regimens are eligible to cure H. pylori infection. However, none of the medication combinations were able to cure H. pylori infection in more than 85% of the patients assessed by intention-to-treat. The countries in which the studies were performed also had a significant impact on eradication rates. PMID- 10383519 TI - Short-term triple therapy with lansoprazole 30 mg or 60 mg, amoxycillin and clarithromycin to eradicate Helicobacter pylori. AB - BACKGROUND: We investigated the efficacy of 30 vs. 60 mg lansoprazole daily in a 1-week triple therapy for eradication of Helicobacter pylori in a prospective randomized study. METHODS: Two hundred and fifteen consecutive out-patients with peptic ulcer disease or non-ulcer dyspepsia, in whom H. pylori infection was confirmed by histology and/or a urease biopsy test, were randomly assigned to a 1 week treatment with either 15 mg lansoprazole b.d. (LAC15 group) or 30 mg lansoprazole b.d. (LAC30 group) in combination with 1 g amoxycillin b.d. and 500 mg clarithromycin b.d. RESULTS: Eradication of H. pylori was successful in 87% (per protocol) and 82% (intention-to-treat) of the patients with LAC15 and in 94% (per protocol) and 87% (intention-to-treat) of the patients with LAC30. The difference was not significant. In both treatment groups, all peptic ulcers were healed at the check-up. Adverse effects were seen in 11 patients of the LAC15 group and 10 patients of the LAC30 group: they caused discontinuation of the therapy in four of the LAC15 group and two patients of the LAC 30 group. CONCLUSIONS: A 7-day triple therapy using lansoprazole (LAC15) is an efficient and economical regimen for the eradication of H. pylori. PMID- 10383520 TI - Comparison of two 3-day Helicobacter pylori eradication regimens with a standard 1-week regimen. AB - BACKGROUND: The duration of Helicobacter pylori eradication regimens has decreased to 1 week with cure rates of over 90%. This can be attributed to the use of triple drug regimens including potent inhibitors of gastric acid secretion and clarithromycin. There is no theoretical reason why shorter regimens should not be possible. AIM: To compare two 3-day, low-dose, twice daily regimens with 1 week of omeprazole 20 mg b.d., clarithromycin 250 mg b.d., and metronidazole 400 mg b.d. (OCM) METHODS: Outpatients referred for gastroscopy were screened by biopsy urease test. H. pylori-positive patients were randomized to receive either lansoprazole 30 mg b.d., tri-potassium dicitrato bismuthate one tablet b.d., clarithromycin 250 mg b.d., and amoxycillin 1 g b.d. for 3 days (LTdbCA), or ranitidine bismuth citrate 400 mg b.d., clarithromycin 250 mg b.d. and amoxycillin 1 g b.d. for 3 days (RbcCA) or omeprazole 20 mg b.d., clarithromycin 250 mg b.d. and metronidazole 400 mg b.d. for 1 week (OCM). They were not pre treated with a gastric acid inhibitor. After 8 weeks, H. pylori status was assessed by 13C urea breath test. RESULTS: 974 out of 1114 patients referred for gastroscopy were screened by biopsy urease test. 140 patients were not screened either because they were anticoagulated or for technical reasons. 334 patients were H. pylori-positive: 154 were excluded mostly because of allergy to penicillin and personal reasons but 180 were randomized to treatment All regimens were well tolerated. For LTdbCA (n=60), RbcCA (n=59), and OCM (n=61) the H. pylori cure rates (95% CI) were 23% (12-34), 14% (5-23) and 87% (79-95), respectively, using intention-to-treat analysis and 25% (14-36), 15% (6-24) and 88% (80-96), respectively, if analysed per protocol. OCM was significantly superior to LTdbCA and RbcCA (P < 0.001) but there was no significant difference between regimens LTdbCA and RbcCA. CONCLUSIONS: OCM is an extremely effective H. pylori eradication regimen. The 3-day regimens tested both have poor cure rates. Pre-treatment with a proton pump inhibitor, higher doses or more frequent dosing may be necessary to increase the cure rate of short duration regimens. However, this could make them less acceptable than the H. pylori eradication regimens currently available. PMID- 10383521 TI - Effect of ranitidine bismuth citrate on the phospholipase A2 activity of Naja naja venom and Helicobacter pylori: a biochemical analysis. AB - BACKGROUND: Helicobacter pylori has become recognized as a fundamental pathogen in the development of gastritis and peptic ulcer disease. Bismuth compounds in combination with antibiotics are widely used to treat H. pylori associated peptic ulcer disease. METHODS: In this study we measured and analysed the inhibitory effect of ranitidine bismuth citrate (RBC, Pylorid, Tritec) on the activity and kinetics of phospholipase A2 (PLA2) (E.C.3.1.1.4) of commercial cobra (Naja naja) venom and H. pylori (French press lysates) using L-alpha-dipalmitoyl-(2[1 14C]palmitoyl)-phosphatidylcholine as substrate. RESULTS: Our data suggest that RBC might exert a dose-dependent uncompetitive inhibition on PLA2 activity of both H. pylori and Naja naja venom. the inhibitory effect of RBC on the PLA2 activity cannot be abolished by the optimal concentration of calcium (10 mM), indicating its mechanism to be unrelated to the displacement of calcium from the activation site of the enzyme. CONCLUSION: Our results suggest that one of the mechanisms by which bismuth compounds are therapeutically effective in the treatment of H. pylori associated gastritis is by inhibiting the activity of the degradative PLA2 enzyme secreted by H. pylori. As a consequence of the inhibitory action of RBC on PLA2 of the bacteria, the extracellular and/or intracellular phospholipid components of the gastric mucosal barrier are preserved. PMID- 10383522 TI - Randomized trial of omeprazole and clarithromycin combined with either metronidazole or amoxycillin in patients with metronidazole-resistant or susceptible Helicobacter pylori strains. AB - BACKGROUND: The impact of metronidazole resistance on the efficacy of proton pump inhibitor based triple therapies remains unclear. AIM: To study whether metronidazole resistance affects Helicobacter pylori eradication rates in patients treated for 1 week with either omeprazole 20 mg b.d., metronidazole 400 mg b.d. and clarithromycin 250 mg b.d. (OMC), or omeprazole 20 mg b.d., amoxycillin 1000 mg b.d. and clarithromycin 500 mg b.d. (OAC). METHODS: A randomized, single blind, single centre study with parallel groups was conducted. H. pylori positive patients were enrolled in a metronidazole-resistant (MR; MIC > 8 microgram/mL) or a metronidazole-susceptible group (MS; MIC < 4 microgram/mL), as determined by E-test. Within the strata patients were randomized to either OAC or OMC. RESULTS: One hundred and twenty-two patients were included. The per protocol cure rate for OAC was 52 out of 57 (91%) (MS 23 out of 26 (89%); MR 29 out of 31 (94%)) and for OMC 46 out of 55 (84%) (MS 19 out of 22 (86%); MR 27 out of 33 (82%)). CONCLUSIONS: One-week OAC and OMC are effective therapies. OAC and OMC were equally effective in patients with metronidazole-susceptible strains of H. pylori. Using the OMC regimen, neither equality nor significant differences in treatment outcome could be shown between patients with metronidazole-resistant or -susceptible strains of H. pylori. PMID- 10383523 TI - Low-dose intravenous azathioprine may be effective in the management of acute fulminant colitis complicating inflammatory bowel disease. AB - BACKGROUND: The management of acute fulminant colitis unresponsive to intravenous steroids is usually surgical. However, recent evidence suggests that intravenous administration of azathioprine at very high doses may allow more rapid onset of clinical efficacy, although its use has not previously been reported in the emergency situation. AIM: To report the successful use of intravenous azathioprine in the management of acute fulminant colitis complicating both Crohn's disease and ulcerative colitis. METHOD: We initially used intravenous azathioprine because of the refusal of the family of the first patient to accept surgery following failure of conventional medical management. Importantly the azathioprine was successful at the low dose of 3 mg/kg.day, equivalent to standard oral doses. Two subsequent patients demonstrated a similar resolution. All were weaned successfully to oral azathioprine and have remained in long-term endoscopic and histological remission. CONCLUSION: These preliminary data suggest that low-dose intravenous azathioprine may be helpful adjunct therapy in selected cases of severe fulminant colitis. However, the need for close monitoring and daily surgical assessment remains paramount, and a formal trial of low-dose intravenous azathioprine is required before it may be more widely recommended. PMID- 10383524 TI - Gastrointestinal safety and tolerance of ibuprofen at maximum over-the-counter dose. AB - BACKGROUND: Delineation of non-steroidal anti-inflammatory drug (NSAID) gastrointestinal toxicity has largely depended on retrospective epidemiologic studies which demonstrate that lower doses of NSAIDs pose a lower risk of gastrointestinal toxicity. Ibuprofen, a propionic acid NSAID, has, in most such studies, exhibited a favourable profile in terms of gastrointestinal bleeding. Since 1984, ibuprofen has been available as a non-prescription analgesic/antipyretic with a limit of 1200 mg/day for 10 days of continuous use. Trials and spontaneously reported adverse experiences suggest that gastrointestinal symptoms and bleeding are rare. METHODS: This study prospectively evaluated the gastrointestinal tolerability, as compared to placebo, of the maximum non-prescription dose and duration of ibuprofen use in healthy subjects representative of a non-prescription analgesic user population. RESULTS: Gastrointestinal adverse experiences were similar in the placebo and ibuprofen groups (67 out of 413, 16% with placebo vs. 161 out of 833, 19% with ibuprofen). There was no difference between the two groups in the proportion discontinuing due to a gastrointestinal event. Gastrointestinal adverse experiences reported by >/= 1% of subjects were: dyspepsia, abdominal pain, nausea, diarrhoea, flatulence, and constipation. Seventeen (1.4%) subjects had positive occult blood tests: their frequency was comparable between treatments. CONCLUSIONS: When used as directed to treat episodic pain, non-prescription ibuprofen at the maximum dose of 1200 mg/day for 10 days, is well-tolerated. PMID- 10383526 TI - The effect of Helicobacter pylori eradication on gastro-oesophageal reflux. AB - BACKGROUND: Increased prevalence of oesophagitis has been reported following eradication of Helicobacter pylori. We hypothesized that H. pylori eradication might increase gastro-oesophageal acid reflux in patients with reflux oesophagitis. METHODS: Twenty-five consecutive patients (13 male, 12 female) with H. pylori infection and reflux oesophagitis grade I (22 patients) or II (three patients) were enrolled; mean age 49.9 (range 33-75) years. Twenty-four hour intra-oesophageal pH recording was performed before and 12 weeks after eradication of H. pylori, which was achieved using bismuth subnitrate suspension 150 mg q.d.s., oxytetracycline 500 mg q.d.s. and metronidazole 400 mg t.d.s. for 10 days. Eradication was confirmed by 14C-urea breath test 12 weeks after completion of treatment. The patients did not receive acid-suppressive medication. RESULTS: All patients had abnormal gastro-oesophageal reflux before anti-H. pylori treatment. After treatment, there was no significant change in the percentage of total time oesophageal pH < 4 (P=0.46) in the 23 patients in whom the infection had been cured. Nine of the cured patients had increased acid exposure, whereas 14 had decreased acid exposure. No significant change in reflux symptom scores was found. There was no relationship between change in acid exposure and symptom improvement. CONCLUSIONS: Twelve weeks after H. pylori eradication there was no consistent change in gastro-oesophageal acid reflux in patients with mild or moderate reflux oesophagitis. PMID- 10383525 TI - On demand therapy with omeprazole for the long-term management of patients with heartburn without oesophagitis--a placebo-controlled randomized trial. AB - AIM: To observe the natural course of gastro-oesophageal reflux disease (GERD) in patients without oesophagitis following effective symptom relief, and to determine the place of acid pump inhibitor therapy in the long-term management of these patients. METHODS: We investigated the efficacy of on-demand therapy with omeprazole 20 mg or 10 mg, or placebo in a double-blind, randomized multicentre trial. It involved 424 patients with troublesome heartburn without endoscopic evidence of oesophagitis in whom heartburn had been resolved with short-term treatment. Patients were told to take study medication on demand once daily on recurrence of symptoms until symptoms resolved over a 6-month period. They also had access to antacids. The primary efficacy variable was time to discontinuation of treatment, due to unwillingness to continue. RESULTS: According to life-table analysis, after 6 months the remission rates were 83% (95% CI: 77-89%) with omeprazole 20 mg, 69% (61-77%) with omeprazole 10 mg, and 56% (46-64%) with placebo (P < 0.01 for all intergroup differences). The mean (s.d.) number of study medications used per day in these groups was 0.43 (0.27), 0.41 (0.27) and 0.47 (0.27), respectively. The use of antacids was highest in the placebo group and lowest in the omeprazole 20 mg group. Treatment failure was associated with more than a doubling of antacid use, and a deterioration in patient quality of life. CONCLUSIONS: Approximately 50% of patients with heartburn who do not have oesophagitis need acid inhibitory therapy in addition to antacid medication to maintain a normal quality of life. On-demand therapy with omeprazole 20 mg, is an effective treatment strategy in these patients. PMID- 10383527 TI - The influence of Helicobacter pylori on oesophageal acid exposure in GERD during acid suppressive therapy. AB - BACKGROUND: Helicobacter pylori exaggerates the effect of acid suppressive drugs on intragastric pH. It is unknown whether this is relevant for the treatment of GERD. AIM: To compare oesophageal acid exposure and symptoms in H. pylori negative and H. pylori-positive GERD patients during low and profound acid suppression. METHODS: Barrett's oesophagus patients with gastro- oesophageal acid reflux were studied by 24-h oesophageal pH-metry at baseline and during randomized treatment with omeprazole 40 mg b.d. or ranitidine 150 mg b.d. H. pylori status was determined by a serum IgG ELISA. Symptoms were scored on a four graded scale. RESULTS: Of 58 patients, 26 (14 H. pylori-negative, 12 H. pylori positive) were randomized to omeprazole, 32 (16 H. pylori-negative, 16 H. pylori positive) to ranitidine. At baseline, oesophageal acid exposure and symptoms did not differ between H. pylori-negative and H. pylori-positive: mean time proportion pH < 4 per 24 h was 16.1% (95% CI 11.5-23.2) in H. pylori-negative, and 15.8% (11.3-21.4) in H. pylori-positive patients. Omeprazole treatment resulted in a decrease of acid reflux per 24 h from 23.4% (7.9-39.3) to 0.0% (0.0 2.9) in H. pylori-negative, and from 17.3% (8.9-38.8) to 0.1% (0.0-1.7) in H. pylori-positive patients; ranitidine resulted in a decrease from 14.4% (10.5 18.5) to 9.3% (5.6-12.8) in H. pylori-negative, and from 15.1% (9.8-21.0) to 9.0% (3.1-20.1) in H. pylori-positive patients, the difference between H. pylori negative and H. pylori-positive patients being N.S. There was no significant difference between H. pylori-negative and H. pylori-positive patients with respect to erect and supine acid reflux, or symptom scores in both treatment groups. CONCLUSIONS: H. pylori infection does not influence oesophageal acid reflux and symptoms in patients with Barrett's oesophagus, either at baseline or during low as well as profound acid suppressive therapy. We conclude that the dose of acid suppression does not have to be titrated upon H. pylori status in GERD. PMID- 10383528 TI - Altered oesophageal motility following the administration of the 5-HT1 agonist, sumatriptan. AB - BACKGROUND: The 5-HT1 agonist sumatriptan, used in the treatment of migraine, can cause chest pain. AIM: To investigate the effect of a therapeutic dose of sumatriptan (6 mg s.c.) on oesophageal motility. METHODS: In 16 normal healthy subjects aged 19-32 years (9 males), the manometric response of the lower oesophageal sphincter (sleeve sensor), oesophageal body (four sites), stomach and pharynx (to register swallows) to 5 mL water swallows was assessed before and after a subcutaneous injection of either sumatriptan (6 mg) or saline control. Symptoms and ECGs were also monitored. RESULTS: Sumatriptan 6 mg s.c. altered oesophageal motility in all subjects. This was reflected by a significant increase in the amplitude of oesophageal body contractions (change from pre- to 1 h post-injection: sumatriptan 9.9 (2.8, 17.1) mmHg vs. placebo -0.8 (-4.2, 2.6) mmHg, difference 10.8 (4.4, 17.1) mmHg; P=0.003) and a transient increase in lower oesophageal sphincter pressure (change from pre- to 5 min post-injection: sumatriptan 10.9 (5.2, 16.6) mmHg vs. placebo 5.1 (1.8, 8.4) mmHg, difference 5.8 (-0.7, 12.3) mmHg; P=0.08). Sumatriptan had no effect on the velocity of propagation of oesophageal contractions (change from pre- to 1 h post-injection: sumatriptan -0.1 (-0.3, 0.1) cm/s vs. placebo -0.1 (-0.3, 0.0) cm/s, difference 0.1 (-0.1, 0.2) cm/s; P = 0.40). One subject experienced chest symptoms following sumatriptan and, although motility was altered, this did not reach pathological levels. No ECG abnormalities were observed. CONCLUSION: Sumatriptan (6 mg s.c.) significantly alters oesophageal motor function without affecting the ECG. It is therefore possible that sumatriptan-induced chest symptoms may have an oesophageal origin. The evaluation of similar therapeutic agents for migraine on oesophageal function may be justified. PMID- 10383529 TI - Post-prandial intragastric and duodenal acidity are increased in patients with chronic pancreatitis. AB - OBJECTIVES: Patients with chronic pancreatitis and exocrine insufficiency have lower intraduodenal pH compared to controls. It has been assumed that abnormal low intraduodenal pH in these patients not only results from impaired pancreatic bicarbonate secretion but also from an increased gastric acid load to the duodenum. METHODS: We have tested this hypothesis by combined intragastric and intraduodenal 24 h pH monitoring in nine chronic pancreatitis patients with exocrine pancreatic insufficiency and nine healthy control subjects during standardized test conditions. Postprandial gastrin and cholecystokinin release were also determined. RESULTS: Median 24-h intraduodenal pH (5.90 vs. 6.00) and intragastric pH (1.60 vs. 1.70) were not significantly different between patients and controls. However, in the 2-h postprandial periods intraduodenal pH was below five for a significantly higher percentage of time in chronic pancreatitis patients compared to controls (lunch: 14.5% vs. 0.17%, P=0.011; dinner: 24.1% vs. 5.75%, P=0.05). The post-dinner intragastric pH was below three for a significantly higher percentage of time in chronic pancreatitis patients vs. controls (72.2 vs. 48.9%, P=0.04). Postprandial gastrin release was not significantly different between the two groups. Postprandial secretion of cholecystokinin (CCK), as enterogastrone, was significantly (P < 0.01) reduced in chronic pancreatitis patients (78 +/- 13 pmol/L, 120 min) compared to controls (155 +/- 14 pmol/L, 120 min). CONCLUSIONS: Median intraduodenal and intragastric pH are not significantly decreased in patients with chronic pancreatitis and exocrine insufficiency but the postprandial time with an acidic pH in the duodenum (pH < 5) and in the stomach (pH < 3) is significantly (P 7.5 g/day during a placebo period were randomly assigned either to the minimicrosphere/microsphere treatment sequence or vice versa. The primary end point was the coefficient of fat absorption, which was calculated from fat excretion and fat intake during the course of a standardized diet. Stool weight, clinical symptoms and the safety of the preparations were also evaluated. RESULTS: Thirty-seven patients entered the study, of whom 23 fulfilled the criteria for the crossover period. In the per protocol analysis (n=18), the 90% confidence intervals for the coefficient of fat absorption of both crossover periods lay entirely within the equivalence range (P=0.02). The intention-to treat analysis revealed similar results, but the equivalence range was slightly missed (P=0.07). Similar results were obtained for the secondary parameters and the reported adverse events. CONCLUSIONS: Pancreatin minimicrospheres have been shown to be equally effective as microspheres in improving the coefficient of fat absorption in patients with exocrine insufficiency due to chronic pancreatitis. PMID- 10383532 TI - Double-blind comparison of lansoprazole 15 mg, lansoprazole 30 mg and placebo as maintenance therapy in patients with healed duodenal ulcers resistant to H2 receptor antagonists. AB - BACKGROUND: Maintenance antisecretory therapy is often used to prevent duodenal ulcer recurrence and control symptoms. This study compared the efficacy and safety of lansoprazole 15 mg and 30 mg daily with placebo in preventing ulcer recurrence in patients with a recent history of duodenal ulcer disease. METHODS: Fifty-six patients were treated with either lansoprazole 15 mg, 30 mg or placebo o.m. RESULTS: Within 1 month of study initiation, 27% (four out of 15) of placebo treated patients experienced ulcer recurrence as compared to 13% (two out of 15) and 6% (one out of 18) of lansoprazole 15 mg and 30 mg treated patients, respectively. Median time to first ulcer recurrence was > 12 months in lansoprazole patients. At Month 12, significantly (P < 0.001) more lansoprazole 15 mg patients (70%) and lansoprazole 30 mg patients (85%) remained healed. Eighty-two per cent of lansoprazole 15 mg and 76% of lansoprazole 30 mg patients remained asymptomatic during the entire study period. All placebo patients became symptomatic, experienced ulcer recurrence, or withdrew from the study by month six. The incidence of adverse events was comparable among the three treatment groups. CONCLUSIONS: Lansoprazole safely and effectively reduces duodenal ulcer recurrence and ulcer-related symptoms. PMID- 10383533 TI - Octreotide in refractory functional epigastric pain with nutritional impairment- an open study. AB - AIM: To test the therapeutic efficacy of octreotide administered subcutaneously for the relief of chronic refractory epigastric pain severe enough to provoke nutritional impairment. SUBJECTS AND METHODS: Seventeen patients were enrolled in an open trial. Epigastric pain had lasted from 1 to 8 years (median: 5 years), following anti-reflux surgery in eight patients. Median weight loss was 10% (range 10-15). The initial dose of octreotide was 50 microgram b.d, adjusted during the follow-up visits which were scheduled for months 1, 3, 6, 8, 10, 12 and every 3 months. At each visit, overall symptomatic improvement, frequency and intensity of symptoms were checked on a 10-cm visual analogic scale. RESULTS: At month 1, a progressive improvement of pain intensity was reported in 15 of the 17 patients, while octreotide was a therapeutic failure in two. In four out of 15, the daily dose of octreotide was increased to 100 microgram b.d. In these 15 patients, median follow-up was 7 months (3-27). The symptomatic benefit was maintained in each patient at month 3, with a median weight gain of 3.5 kg.2-5 An attempt to stop octreotide led to recurrence of symptoms in 2-3 days which were as intense as before the treatment. The 11 patients followed-up for at least 6 months reported persistent improvement of symptoms with octreotide and a median weight gain of 4 kg.3-7 Four patients were followed up for more 11-27 months: octreotide was withdrawn gradually in two who remained asymptomatic. Six of the 17 patients experienced minor side-effects, but none developed biliary sludge. CONCLUSIONS: This open study suggests that octreotide could be a promising alternative treatment when all others fail in refractory chronic functional epigastric pain severe enough to limit food intake and to induce nutritional impairment. These results must be tested by a placebo-controlled study. PMID- 10383534 TI - Nocturnal gastric acid breakthrough on proton pump inhibitors. PMID- 10383537 TI - The interface between clinical and laboratory pharmacology. PMID- 10383536 TI - The management of Pneumocystis carinii pneumonia. PMID- 10383538 TI - Relative bioavailability of sodium cromoglycate to the lung following inhalation, using urinary excretion. AB - AIMS: To determine if a urinary excretion method, previously described for salbutamol, could also indicate the relative bioavailability of sodium cromoglycate to the lung following inhalation from a metered dose inhaler. Method Inhaled (INH), inhaled+oral charcoal (INHC), oral (ORAL) and oral+oral charcoal (ORALC) 20 mg doses of sodium cromoglycate were given via a randomised cross-over design to 11 healthy volunteers trained on how to use a metered dose inhaler. Urine samples were collected at 0.0, 0.5, 1.0 and up to 24 h post dosing and the sodium cromoglycate urinary concentration was measured using a high performance liquid chromatographic method. RESULTS: No sodium cromoglycate was detected in the urine up to 24 h following ORALC dosing. A mean (s.d.) of 3.6 (4.3) microg, 10.4 (10.9) microg and 83.7 (71.1) microg of the ORAL dose was excreted, in the urine, during the 0.5, 1.0 and 24 h post dose collection periods, respectively. Following INH dosing, the renal excretion was significantly higher (P<0.01) with 32.9 (14.5) microg, 61.2 (28.3) microg and 305.6 (82.3) microg excreted, respectively. The SCG excreted at 0.5, 1.0 and 24 h collection periods following INHC dosing were 26.3 (8.4) microg, 49.3 (18.1) microg and 184.9 (98.4) microg, respectively. There was no significant difference between the excretion rate of sodium cromoglycate following INHC when compared with INH dosing in the first 0.5 and 1.0 h. CONCLUSIONS: The urinary excretion of sodium cromoglycate in the first 0.5 h post inhalation can be used to compare the relative lung deposition of two inhaled products or of the same product using different inhalation techniques. This represents the relative bioavailability of sodium cromoglycate to the lung following inhalation. Similar 24 h urinary excretion of sodium cromoglycate can be use to compare the total dose delivered to the body from two different inhalation products/inhalation methods. This represents the relative bioavailability of sodium cromoglycate to the body following inhalation. Because of the lack of difference between the INH and INHC in the first 0.5 h, the use of activated charcoal is not necessary when this method is used to compare the relative lung bioavailability of different products or techniques. PMID- 10383539 TI - Systemic availability of budesonide after nasal administration of three different formulations: pressurized aerosol, aqueous pump spray, and powder. AB - AIMS: The present study was undertaken to determine the absolute systemic availability of budesonide from three different devices for nasal administration: pressurized aerosol, aqueous pump spray, and powder. METHODS: Sixteen healthy, non-smoking, volunteers participated in this open, randomized, and crossover study. All subjects received budesonide as an intravenous dose of 400 microg, and as three, single-dose, intranasal administrations: pressurized aerosol 800 microg, aqueous pump spray 400 microg, and powder 800 microg. Blood was sampled for 10 h after each administration and budesonide was assayed in plasma by liquid chromatography plus mass spectrometry. RESULTS: The mean [95% CI] systemic availability of budesonide with reference to the metered dose was: 13 [10; 15]%, 29 [23; 37]%, and 20 [16; 23]%, and the maximum plasma concentration (Cmax) was attained at (tmax) 2.0, 0.7, and 0.4 h after administration for the pressurized aerosol, aqueous pump spray, and powder, respectively. CONCLUSIONS: The uptake of budesonide was more rapid and more complete, and the systemic availability of the drug was significantly higher from the aqueous pump spray and powder than from the pressurized aerosol. PMID- 10383540 TI - Microsomal prediction of in vivo clearance of CYP2C9 substrates in humans. AB - AIMS: To assess the utility of human hepatic microsomes for predicting in vivo intrinsic clearance (CLint ) via the use of four cytochrome P450 2C9 substrates: phenytoin, tolbutamide (S)-ibuprofen (two pathways) and diclofenac, and to examine the role of exogenous albumin within the microsomal incubation. METHODS: V max, Km and CLint (defined as V max/Km ratio) were estimated under initial rate conditions for five pathways of metabolism in a bank of 15 human hepatic microsomal samples and were scaled to in vivo units using the microsomal protein index. Non-metabolic related binding in microsomes was measured for phenytoin and tolbutamide in the presence and absence of albumin. RESULTS: Microsomal CLint values differed by over two orders of magnitude, with the means ranging from 0.18 (phenytoin) to 40.70 (diclofenac) microl min-1 mg-1 microsomal protein. When these data were scaled and compared with published in vivo studies a similar rank order was obtained, however, the actual CLint tended to be underpredicted. While the in vivo unbound Km for phenytoin, 1-5 micron is substantially lower than the value determined in microsomes based on total concentrations (56 micron), correction for the in vitro binding reduces this value to 20 micron and 6 micron in the absence and presence of albumin, respectively. Similar trends were seen with tolbutamide Km. CONCLUSIONS: An appreciation of the utility of in vitro prediction can be best achieved when the range of CLint values predicted from the individual hepatic microsomal samples are compared with the range of individual in vivo CLint values reported in the literature. The degree of underprediction is less evident using the range than the mean data and no consistent advantage in adding albumin to the incubation media is apparent. PMID- 10383541 TI - Pharmacokinetics of gentamicin in 957 patients with varying renal function dosed once daily. AB - AIMS: 1. To determine the population pharmacokinetics of gentamicin in 957 patients with varying renal function dosed once daily. 2. To see if current starting doses for once daily aminoglycoside dosing are appropriate. 3. To test whether calculating creatinine clearance using an adjusted Cockcroft and Gault method (CLCr,adjusted ) was a better predictor of gentamicin clearance than the standard Cockcroft and Gault method (CLCr,unadjusted ). METHODS: Nine hundred and fifty-seven patients were dose-individualized for gentamicin using SeBA-GEN, a Bayesian dosing method. This method returns estimates of the values of gentamicin CL and V d from which the 24 h AUC can be estimated. The goal of therapy was to attain an AUC of 70-100 mg l-1 h depending on the severity of the infection. The population was divided into four groups of differing renal function. Linear regression analysis was performed to determine the relationship between V d and various indices of weight, and gentamicin CL and either CLCr,adjusted or CLCr,unadjusted. RESULTS: The mean V d (+/-s. d.) and CL (+/-s.d.) of gentamicin in our total population were 17.4 (+/-4.1) l and 4.0 (+/-1.8) l h-1, respectively. There was a decrease in V d with reducing renal function when comparing patients with normal renal function and patients with poor renal function. The lower of total body weight (TBW) and lean body weight (LBW), termed dosing weight (DWT), was a slightly better predictor of V d (r2=0.28) than either TBW (r2=0.21) or LBW (r2=0.21). CLCr,adjusted (r2=0.80) was a better predictor of gentamicin CL than CLCr, unadjusted (r2=0.57). CONCLUSIONS: The mean population values of V d and CL of gentamicin dosed once daily are similar to those described by others in relation to multiple daily dosing. Given that previous methods have been based on population values of V d and CL from multiple daily dosing, the currently recommended starting doses for once daily aminoglycoside dosing would seem appropriate. The V d reduced with decreasing renal function, with a maximum of 23% difference between patients with normal and poor renal function. The Cockcroft and Gault method of calculating creatinine clearance does not appear to perform well at low values of serum creatinine concentration. An adjustment of the Cockcroft and Gault method is proposed to allow for this. PMID- 10383542 TI - Pharmacokinetics and haemodynamics of candesartan cilexetil in hypertensive patients on regular haemodialysis. AB - AIMS: The pharmacokinetic profile of candesartan cilexetil might be altered in patients with end-stage renal disease (ESRD). No data are available about the pharmacokinetics and haemodynamics of the angiotensin II receptor antagonist candesartan cilexetil in ESRD patients on regular haemodialysis (HD). METHODS: We performed a repeated dose study (8 mg candesartan cilexetil once daily) in eight male HD patients over a treatment period of 5 days with an additional observation period of 3 days. RESULTS: Pharmacokinetic analysis with nonlinear mixed effects modeling (NONMEM) over the whole treatment period revealed a dependency of the volume of distribution on body weight and of the metabolic clearance on age and body weight in the studied population. No significant drug elimination by HD was observed. The estimated metabolic and intercompartmental clearances were 83 ml min-1 (CV 39%) and 9.9 ml min-1, respectively. The unexplained random variability of the final two compartment model was 30%. In one patient with adult polycystic kidney disease oral clearance decreased during the observation period, attributable to a significant increase in bioavailability. Maximum observed changes in blood pressure were -50/-27+/-14/8 mmHg on day 5 with haemodialysis therapy as compared with changes in blood pressure of -14/-12+/-14/8 mmHg on day 1 without haemodialysis treatment. The observed maximum decrease in systolic blood pressure correlated with the amount of ultrafiltration during the HD session on day 5 (r=0.70, P<0.05). In two patients, one of whom was binephrectomized, severe hypotensive episodes were observed during this HD session. CONCLUSIONS: HD does not influence the elimination kinetics of candesartan. The observed inter- and intraindividual variability of oral clearance and the pronounced influence of HD-induced volume contraction on the haemodynamic effects of candesartan makes it mandatory to carefully monitor HD patients treated with candesartan cilexetil. PMID- 10383543 TI - Pharmacokinetics, induction of anaesthesia and safety characteristics of propofol 6% SAZN vs propofol 1% SAZN and Diprivan-10 after bolus injection. AB - AIMS: In order to avoid the potential for elevated serum lipid levels as a consequence of long term sedation with propofol, a formulation of propofol 6% in Lipofundin(R) MCT/LCT 10% (Propofol 6% SAZN) has been developed. The pharmacokinetics, induction of anaesthesia and safety characteristics of this new formulation were investigated after bolus injection and were compared with the commercially available product (propofol 1% in Intralipid(R) 10%, Diprivan-10) and propofol 1% in Lipofundin(R) MCT/LCT 10% (Propofol 1% SAZN). METHODS: In a randomised double-blind study, 24 unpremedicated female patients received an induction dose of propofol of 2.5 mg kg-1 over 60 s which was followed by standardized balanced anaesthesia. The patients were randomized to receive propofol as Propofol 6% SAZN, Propofol 1% SAZN or Diprivan-10. RESULTS: For all formulations the pharmacokinetics were adequately described by a tri-exponential equation, as the propofol concentrations collected early after the injection suggested an additional initial more rapid phase. The average values for clearance (CL), volume of distribution at steady-state (Vd,ss ), elimination half life (t1/2,z ) and distribution half-life (t1/2, lambda2) observed in the three groups were 32+/-1.5 ml kg-1 min-1, 2. 0+/-0.18 l kg-1, 95+/-5.6 min and 3.4+/ 0.20 min, respectively (mean+/-s.e.mean, n=24) and no significant differences were noted between the three formulations (P >0.05). The half-life of the additional initial distribution phase (t1/2,lambda1 ) in all subjects ranged from 0.1 to 0.6 min. Anaesthesia was induced successfully and uneventfully in all cases, and the quality of induction was adequate in all 24 patients. The induction time did not vary between the three formulations and the average induction time observed in the three groups was 51+/-1.3 s which corresponded to an induction dose of propofol of 2.1+/-0.06 mg kg-1 (mean+/-s.e. mean, n=24). The percentage of patients reporting any pain on injection did not vary between the formulations and was 17% for the three groups. No postoperative phlebitis or other venous sequelae of the vein used for injection occurred in any of the patients at recovery of anaesthesia nor after 24 h. CONCLUSIONS: From the above results, we conclude that the alteration of the type of emulsion and the higher concentration of propofol in the new parenteral formulation of propofol does not affect the pharmacokinetics and induction characteristics of propofol, compared with the currently available product. Propofol 6% SAZN can be administered safely and has the advantage of a reduction of the load of fat and emulsifier which may be preferable when long term administration of propofol is required. PMID- 10383544 TI - Ursodeoxycholic acid and endothelial-dependent, nitric oxide-independent vasodilatation of forearm resistance arteries in patients with coronary heart disease. AB - AIMS: Ursodeoxycholic acid (UDCA) has cholesterol lowering and anti-inflammatory effects and bile acids are reported to exert vasodilator effects; all of these properties might be considered desirable in a drug used in the treatment of patients with coronary heart disease. We investigated a hypothesis that UDCA may dilate arteries and the mechanism of action. METHODS: We evaluated effects of a 6 week treatment with UDCA in 11 coronary heart disease patients on endothelium dependent (acetylcholine-induced) and -independent (nitroprusside-induced) vasodilatations in forearm vasculature by strain-gauge plethysmography. Healthy individuals (n=14) served as baseline controls. RESULTS: The percentage increase by acetylcholine in the flow of the infused arm relative to the non-infused arm of coronary heart disease patients during the trial remained unaltered, but vasodilatation to NG-monomethyl-l-arginine+acetylcholine was improved by 161+/ 27% with UDCA vs 83+/-22% with placebo (mean difference 91% [95% CI 35%, 147%], P=0.016). CONCLUSIONS: Six weeks' UDCA therapy improved endothelium-dependent nitric oxide-independent vasodilatation, which might maintain arterial flow in coronary heart disease patients under conditions of impaired nitric oxide production. PMID- 10383545 TI - Management of postoperative pain in abdominal surgery in Spain. A multicentre drug utilization study. AB - AIMS: Postoperative pain is common in hospital-admitted patients. Its management is determined by different therapeutic traditions and by the attitudes of health professionals in each hospital. The aim of this study was to describe the patterns of prescription and administration of analgesic drugs used for postoperative pain after abdominal surgery in Spanish hospitals, to know the prevalence and the severity of postoperative pain, and to determine the extent of variability in the management of postoperative pain among the participating centres. METHODS: The study was a multicentre descriptive cross-sectional drug utilization study in 12 Spanish hospitals. The subjects were an unselected sample of consecutive patients undergoing abdominal surgery, admitted between October 1994 and January 1995. For each patient, information about the surgical procedure and the use of analgesics was prospectively collected. The severity of postoperative pain was assessed during the first day after surgery by means of a six-category (none, mild, moderate, severe, very severe, and unbearable) rating scale and a visual analogue scale (VAS). RESULTS: Nine hundred and ninety-three patients (547 men) were included. The most common surgical procedures were inguinal hernia repair (315, 32%), cholecystectomy (268, 27%), appendectomy (140, 14%), bowel resection (137, 14%), and gastric surgery (58, 6%). Fifty-nine percent of patients (587) received nonopioid analgesics only, 9% (89) received opioid analgesics only, and 27% (263) received both opioid and nonopioid analgesics. The most frequently administered drugs were metamizole (667 patients) and pethidine (213 patients). Although in the majority of medical orders the administration of analgesics was scheduled at regular time intervals, the majority of actual doses were given 'as-needed'. The average administered daily doses of all analgesics were lower than those prescribed. Thirty-eight percent (371/967) of patients rated their maximum pain on the first day as severe to unbearable. Wide interhospital variability was recorded in the surgical procedures which had been performed, in the analgesics used, and also in the pain scores referred by patients. The percentage of patients in each centre who suffered severe to unbearable pain varied from 22 to 67%. CONCLUSIONS: In Spain many patients still suffer severe pain after abdominal surgery, and this seems to be due to an inadequate use of analgesics. Wide interhospital variability in the management of postoperative pain and in its prevalence was also recorded. PMID- 10383546 TI - The efficacy of a novel adenosine agonist (WAG 994) in postoperative dental pain. AB - AIMS: To determine the comparative efficacy of a new novel adenosine agonist (WAG 994) in postoperative pain after third molar surgery. METHODS: One hundred and twenty-two patients with postoperative pain after third molar surgery were randomised in a placebo double-blind trial with an active control group. In the early postoperative period patients received either a single dose of WAG 994 1 mg, ibuprofen 400 mg or matched placebos. Pain intensity score was recorded on serial visual analogue scales over a 6 h investigation period. Similarly, pain relief was completed on a 4 point categorical scale at each evaluation point. Patients had access to escape analgesic and if these were taken, the time and dosage were recorded. A sparse sampling technique was used to investigate the relationship between analgesic effects and plasma concentrations of WAG 994. RESULTS: All three treatment groups were matched for various demographic variables. For all efficacy measures, WAG 994 was not significantly different from placebo (P >0.05), whereas ibuprofen 400 mg was significantly superior to placebo (P<0.001). No significant relationships (P<0.05) were found between WAG 994 pharmacokinetic variables and efficacy measures. Conclusion WAG 994, an adenosine agonist, did not show efficacy in the management of postoperative pain after third molar surgery. Although this pain responds well to nonsteroidal anti inflammatory drugs, it appears to be resistant to compounds that interact with purinergic receptors. PMID- 10383547 TI - A prospective study of adverse drug reactions in hospitalized children. AB - AIMS: There are few publications of adverse drug reactions (ADRs) among paediatric patients, though ADR incidence is usually stated to be higher during the first year of life and in male patients. We have carried out a prospective study to assess the extent, pattern and profile risk for ADRs in hospitalized patients between 1 and 24 months of age. METHODS: An intensive events monitoring scheme was used. A total of 512 successive admissions to two medical paediatric wards (47 beds) were analysed. The hospital records were screened daily during two periods (summer, 105 days and winter, 99 days), and adverse clinical events observed were recorded. RESULTS: A total of 282 events were detected; of these, 112 were considered to be manifestations of ADRs. The cumulative incidence was 16.6%, no differences being observed between periods. Although there were no differences between patients under and over 12 months of age, risk was found to be significantly higher among girls compared with boys (RR=1.66, 95% CI 1.03 2.52). The gastro-intestinal system was most frequently affected. The therapeutic group most commonly implicated was anti-infective drugs and vaccines (41.5%). The ADRs were mild or moderate in over 90% of cases. A consistent relationship was noted between the number of drugs administered and the incidence of ADRs. CONCLUSIONS: Hospitalized patients exhibited an ADR risk profile that included female sex and the number of drugs administered. No particular age predisposition was observed. The most commonly prescribed drugs are those most often implicated in ADRs in paediatric patients. PMID- 10383548 TI - Signalling possible drug-drug interactions in a spontaneous reporting system: delay of withdrawal bleeding during concomitant use of oral contraceptives and itraconazole. AB - AIMS: In spontaneous adverse drug reaction reporting systems, there is a growing need for methods facilitating the automated detection of signals concerning possible adverse drug reactions. In addition, special attention is needed for the detection of adverse drug reactions resulting from possible drug-drug interactions. We describe a method for detecting possible drug-drug interactions using logistic regression analysis to calculate ADR reporting odds ratios. METHODS: To illustrate this method, we analysed the adverse drug reaction 'delayed withdrawal bleeding' resulting from a possible interaction between itraconazole and oral contraceptives in reports received by the Netherlands Pharmacovigilance Foundation LAREB between 1991 and 1998. RESULTS: In total 5,503 reports were included in the study. The odds ratio, adjusted for year of reporting, age and source of the reports, for a delayed withdrawal bleeding in women who used both drugs concomitantly compared with women who used neither oral contraceptives, nor itraconazole, was 85 (95% CI: 32-230). CONCLUSIONS: Since spontaneous reporting systems can only generate signals concerning possible relationships, this association needs to be analysed by other methods in more detail in order to determine the real strength of the relationship. This approach might be a promising tool for the development of procedures for automated detection of possible drug-drug interactions in spontaneous reporting systems. PMID- 10383549 TI - Clinical pharmacy interventions by community pharmacists during the dispensing process. AB - AIMS: To evaluate the professional contact between the community pharmacist and general practitioner during the dispensing process on issues other than the legality or simple clarification of the prescription. METHODS: Fourteen community pharmacists from five adjacent localities completed details of each clinical pharmacy intervention during 1 week of each month for a period of 1 year. Each week of the month was randomly selected. When a community pharmacist had to contact the prescriber, during the dispensing of a prescription, the following data were recorded: brief patient details, the prescribed drug therapy, the reason for intervention, the outcome and the time taken. The main outcome measures were the type and nature of each intervention, the BNF category of the drug involved and the time taken. A multidisciplinary clinical panel assessed the potential of each intervention to alter the outcome of the patient's clinical management and to prevent a drug related hospital admission. These assessments were ranked between 0 and 10 (100% confident). RESULTS: During a period covering 1 week per month over 1 year, 1503 clinical pharmacy interventions were made out of 201 000 items dispensed. When normalized for the dispensing volume of each community pharmacy the lower the number of items dispensed then the greater was the percentage of interventions (P=0.013). The clinical panel decided that between 19 (0.01% of the total items dispensed) and 242 (0.12%) interventions may have prevented a drug-related hospital admission, 71 (0.04%) to 483 (0.24%) could have prevented harm whilst 103 (0.05%) to 364 (0.18%) had the potential to improve the efficacy of the intended therapeutic plan. The panel also decided that 748 (0.37%) interventions improved the clinical outcome and could have saved a visit to or by the general practitioner. Conclusion Clinical pharmacy provided by a community pharmacist during the dispensing process has the potential to provide a valuable contribution to health care. PMID- 10383550 TI - The effect of bosentan on the pharmacokinetics of digoxin in healthy male subjects. AB - AIMS: To investigate the effect of multiple oral dose treatment with the endothelin receptor antagonist bosentan on the pharmacokinetics of digoxin in healthy subjects. METHODS: This was an open-label, randomized, two-way crossover study in 18 evaluable young male subjects. They received, on two occasions which were separated by at least 2 weeks washout period, 0.375 mg digoxin once daily for 13 days following a loading dose of 0.375 mg given twice on the day before the once daily dosing regimen started. On one occasion treatment with 500 mg bosentan twice daily was started on the eighth day of digoxin treatment and continued for 1 week. Serum concentrations of digoxin were determined up to 24 h postdose on day 8 (first day of bosentan treatment) and day 14 (last day of bosentan treatment) of the digoxin treatment period. Plasma concentrations of bosentan were measured at two time points after the first bosentan dose and up to 12 h after the last morning dose of bosentan. Safety was assessed by adverse events, clinical laboratory tests, blood pressure and pulse rate measurements and ECG recordings. RESULTS: Steady-state of digoxin was always achieved after 7 days of treatment. Serum concentrations of digoxin were within the usual therapeutic range. Average steady-state Cmax and Ctr were 2-2.1 microg l-1 and 0.65-0.69 microg l-1, respectively, when given alone. Bosentan did not lead to statistically significant changes in Cmax and Ctr of digoxin. AUC (0,24h) of digoxin, however, was slightly reduced after 1 week of treatment with bosentan. The reduction was 12% on average with a narrow 95% confidence interval of 0-23%. Bosentan pharmacokinetic parameters after 1 week of treatment were as expected with a mean Cmax of 3260 microg l-1 and a mean AUC (0, 12h) of 12 600 microg l-1 h. CONCLUSIONS: Treatment with bosentan 500 mg twice daily for 1 week did not show clinically relevant effects on the pharmacokinetics of digoxin in healthy human subjects PMID- 10383551 TI - Clinical pharmacology and therapeutics in a changing world. PMID- 10383552 TI - Paediatric prescribing: using unlicensed drugs and medicines outside their licensed indications. PMID- 10383553 TI - Target concentration intervention: beyond Y2K. AB - Target concentration intervention (TCI) is proposed as an alternative conceptual strategy to therapeutic drug monitoring (TDM). It is argued that the idea of a therapeutic range has limited the interpretation of measured drug concentrations and diminished the anticipated clinical benefit to patients by use of an oversimplified pharmacodynamic model. TCI on the other hand embraces pharmacokinetic and pharmacodynamic concepts and uses the idea of a target effect and associated target concentration to make rational individual dose decisions. PMID- 10383554 TI - Status of new medicines approved by the European Medicines Evaluation Agency regarding paediatric use. AB - AIMS: To evaluate the activity of the European Medicines Evaluation Agency with regard to the registration for paediatric use of new medicines granted a marketing authorization. METHODS: European Public Assessment Reports published on the Internet from January 95 until April 98 have been analysed using the browser Microsoft Explorer and the software Adobe Acrobat Reader. RESULTS: Of the 45 new substances licensed since January 95, 29 (64%) were of possible use in children but only 10 were licensed for paediatric use. For the 19 drugs of possible use in children, but not approved for such a use, in nine instances (47%) their summary of product characteristics reported that their use in children has not been established. CONCLUSIONS: A change of practice by pharmaceutical companies and regulatory authorities is imperative so that children are not precluded from having the same rights to medicines as adults. PMID- 10383555 TI - Transaminase elevation on placebo during phase I trials: prevalence and significance. AB - AIMS: To evaluate the prevalence of transaminase elevation on placebo during Phase I trials. METHODS: Retrospective review of pooled transaminase data collected on placebo during 13 Phase I trials in 93 healthy volunteers hospitalized for 14 days, with determination of the prevalence of abnormally high values. RESULTS: 20.4% of the 93 subjects showed at least one ALT value above the upper limit of the normal range (ULN), and 7.5% had at least one value twice ULN. CONCLUSIONS: Laboratory safety results of Phase I trials should be interpreted with caution, in the light of data on placebo, to avoid premature discontinuation of the development of safe drugs wrongly believed to be hepatotoxic. PMID- 10383556 TI - The prevalence and incidence of medical conditions in healthy pharmaceutical company employees who volunteer to participate in medical research. AB - AIMS: Although clinical research in healthy volunteers is commonly performed there have been few studies of the value of the medical screening of subjects. The aim of this study was to investigate the prevalence and incidence of medical conditions found during the medical screening of 'healthy' subjects employed in a pharmaceutical company who volunteered to participate in medical research. METHODS: This was a retrospective study of the medical notes of all the subjects who volunteered for membership of the Zeneca Clinical Pharmacology Unit's healthy volunteer panel over a 4 year period from 1990 to 1994. The prevalence of medical conditions found at presentation was determined. The incidence of medical conditions during the 4 year observation period was also ascertained. Medical screening included a full medical history and examination, clinical chemistry, haematology and urinalysis screens, pulmonary function tests, ECGs, 24 h ambulatory cardiac monitoring and a request for information from the volunteer's General Practitioner. RESULTS: Prevalence-1293 subjects volunteered to join the panel of which 156 subjects (12%) were not accepted at presentation including 141 (10. 9%) for medical reasons. The most medical common reasons were; previously diagnosed medical conditions (3.3%), cardiovascular abnormalities (1.9%), abnormal liver function tests (1.9%), anaemia (1.2%), hyperlipidaemia (1.1%), excess alcohol intake (0.6%) and thyroid disease (0.5%). Incidence-36 of the 1137 volunteers (0.8% per year) accepted onto the panel subsequently developed medical conditions of which the most common were; anaemia (0.29% per year), cardiovascular abnormalities (0.13% per year) and vasovagal syncope (0.13% per year). CONCLUSIONS: This study demonstrates the importance of medical screening before healthy volunteers participate in clinical research. PMID- 10383557 TI - Netilmicin pharmacokinetics in Hong Kong Chinese cancer patients. AB - OBJECTIVES: To study the pharmacokinetics of netilmicin in Chinese haematology oncology patients and to determine the pharmacokinetic differences, if any, between this patient subpopulation of Chinese and Caucasians. METHODS: A prospective study was carried out in the adult oncology unit of a major hospital in Hong Kong. During a 6 week period in 1997, all patients commencing on netilmicin therapy were monitored; the patients' demographics, clinical status, netilmicin dose and regimen, and drug administration/blood sampling time were collected. Pharmacokinetic parameters were generated using the USC*PACK package based on specifics of the patients themselves and Caucasians matched for the same patients' parameters using the Bayesian alogrithms. RESULTS: A total of 22 patients were enrolled into the study. Twenty-nine sets of levels were drawn, but only 25 sets from 18 patients (86%) were interpretable. The predicted peak (7.47+/-1.46 microg ml-1 ) and trough levels (1.39+/-0.96 microg ml-1 ) generated by USC*PACK were found to be significantly higher than the levels observed (6.01+/-1.14 microg ml-1 and 0.93+/-0.71 microg ml-1, respectively). Netilmicin clearance, volume of distribution and rate of elimination were all significantly higher in this Chinese subpopulation than those predicted for matched Caucasians. Conclusion Alterations in the netilmicin pharmacokinetics observed in our study population might be related to the disease state and/or ethics of the study patient population. Direct application of Caucasian based population pharmacokinetic parameters to this subgroup of Chinese patients may not be appropriate and may result in underdose. PMID- 10383558 TI - Morphine and morphine-6-glucuronide in the plasma and cerebrospinal fluid of children. AB - AIMS: To measure morphine and morphine-6-glucuronide in the plasma and cerebrospinal fluid of children following a single intravenous dose of morphine. METHODS: Twenty-nine paired samples of cerebrospinal fluid and plasma were collected from children with leukaemia undergoing therapeutic lumbar puncture. An intravenous dose of morphine was administered at selected intervals before the procedure. Concentrations of morphine and morphine-6-glucuronide (M6G) were measured in each sample. Morphine was measured using a specific radioimmunoassay (r.i.a.) and M6G was measured using a novel enzyme-linked immunosorbent assay (ELISA). RESULTS: The ELISA for measuring M6G was highly sensitive. The intra-and interassay variations were less than 15%. Using a two-compartment model for plasma morphine, the area under the curve to infinity (AUC, 7143 ng ml-1 min), volume of distribution (3.6 l kg-1 ) and elimination half-life (88 min) were comparable with those reported in adults. Clearance (35 ml min-1 ) was higher than that in adults. Morphine-6-glucuronide was readily synthesized by the children in this study. The elimination half-life (321 min) and AUC (35507 ng ml 1 min) of plasma M6G were much greater than those of morphine. CONCLUSIONS: Extensive metabolism of morphine to M6G in children with cancer has been demonstrated. These data provide further evidence to support the importance of M6G accumulation after multiple doses. There was no evidence that morphine passed more easily into the CSF of children than adults. PMID- 10383559 TI - Population pharmacokinetics of methadone in opiate users: characterization of time-dependent changes. AB - AIMS: Although methadone is widely used to treat opiate dependence, guidelines for its dosage are poorly defined. There is increasing evidence to suggest that a strategy based on plasma drug monitoring may be useful to detect non-compliance. Therefore, we have developed a population-based pharmacokinetic (POP-PK) model that characterises adaptive changes in methadone kinetics. METHODS: Sparse plasma rac-methadone concentrations measured in 35 opiate-users were assessed using the P-Pharm software. The final structural model comprised a biexponential function with first-order input and allowance for time-dependent change in both clearance (CL) and initial volume of distribution (V ). Values of these parameters were allowed to increase or decrease exponentially to an asymptotic value. RESULTS: Increase in individual values of CL and increase or decrease in individual values of V with time was observed in applying the model to the experimental data. CONCLUSIONS: A time-dependent increase in the clearance of methadone is consistent with auto-induction of CYP3A4, the enzyme responsible for much of the metabolism of the drug. The changes in V with time might reflect both up- and down-regulation of alpha1-acid glycoprotein, the major plasma binding site for methadone. By accounting for adaptive kinetic changes, the POP-PK model provides an improved basis for forecasting plasma methadone concentrations to predict and adjust dosage of the drug and to monitor compliance in opiate-users on maintenance treatment. PMID- 10383560 TI - The nasal airways response in normal subjects to oxymetazoline spray: randomized double-blind placebo-controlled trial. AB - AIMS: The effects of a single dose of oxymetazoline nasal spray on nasal patency have been compared with placebo using three separate measuring systems in normal subjects. METHODS: The study was a placebo-controlled, randomised double-blind crossover trial. Subjects without ear, nose or throat disease and with resting nasal airways resistance >0.15 Pa s cm-3 were selected so that a fall in airways resistance could be detected. Nasal airways resistance (NAR) was measured by NR6 2 rhinomanometer. Acoustic rhinometry (SR-2000 rhinometer) provided the sum of the minimum cross-sectional areas (tMCA) and volume (tVOL) of the left and right nasal cavities. Symptoms of congestion were assessed on a visual analogue scale (CON, range 0-100). Measurements were made for 60 min before and for 120 min after bilateral administration of oxymetazoline nasal spray (0.9 mg) or placebo (0.9% saline). Crossover occurred 7-21 days later. Results for all measures were analysed as change from average baseline value by trapezoidal AUC, and statistical significance was tested by 2-way anova. RESULTS: NAR, tMCA, tVOL and CON did not change after placebo, but NAR and CON fell and tMCA and tVOL increased significantly at all timepoints after oxymetazoline. NAR_AUC, tVOL_AUC, tMCA_AUC were significantly different between placebo and oxymetazoline (P<0.001) as was CON_AUC (P=0.012). The day-to-day intraindividual repeatability of baseline NAR tMCA and tVOL was <10%. CONCLUSIONS: Normal subjects can be used to detect the effects of nasally vasoactive drugs with a variety of complementary systems, with the advantages of easy subject recruitment and low variability. PMID- 10383561 TI - Comparison of antiplatelet activity of microencapsulated aspirin 162.5 Mg (Caspac XL), with enteric coated aspirin 75 mg and 150 mg in patients with atherosclerosis. AB - AIMS: A new formulation, low dose microencapsulated aspirin, permits slow absorption of aspirin and presystemic acetylation of platelet cyclo-oxygenase within the portal circulation, potentially avoiding deleterious effects on gastric and systemic prostaglandin synthesis. The objective of this study was to determine whether the administration of microencapsulated aspirin was as effective as enteric coated (EC) aspirin as an inhibitor of platelet function in patients with atherosclerosis. METHODS: One hundred and four patients were enrolled and randomised after a run in period of at least 14 days on aspirin EC 75 mg (day 0), to receive either microencapsulated aspirin 162.5 mg (n=34), aspirin EC 150 mg (n=36) or continue on aspirin EC 75 mg (n=34) for 28 days. Serum thromboxane B2 and collagen-induced platelet aggregation and release of 5 hydroxytryptamine (EC50 values) were measured on days 0 and 28. Aggregation/release EC50s were then repeated in the presence of a large dose of aspirin added in vitro to determine the EC50 at the maximum level of platelet inhibition. RESULTS: Median thromboxane B2 levels were low after 14 days run-in therapy with aspirin EC 75 mg, but significant further reductions were seen on day 28 in patients randomised to microencapsulated aspirin 162.5 mg (P=0.0368) and aspirin EC 150 mg (P=0.0004) compared with those remaining on aspirin EC 75 mg. Median EC50 s on day 28 showed small but significant increases from baseline (day 0) in aggregation in patients randomised to microencapsulated aspirin 162.5 mg (0.62-0.85, P=0.0482) and in both aggregation and release in patients randomised to aspirin EC 150 mg (0.95-1.20, P=0.0002, 8.4-11.7, P<0. 0001, respectively) signifying enhanced antiplatelet activity. No changes were seen in patients continuing on aspirin EC 75 mg. Results following addition of high dose aspirin in vitro suggest that mechanisms other than thromboxane synthesis may be operative in the long term effects of microencapsulated aspirin 162.5 mg and aspirin EC 150 mg over aspirin EC 75 mg. CONCLUSIONS: The results show good inhibition of thromboxane B2 synthesis and subsequent platelet activity by all preparations of aspirin, although both microencapsulated aspirin 162.5 mg and aspirin EC 150 mg are slightly more effective than aspirin EC 75 mg. A randomised trial is now required to determine whether microencapsulated aspirin is associated with fewer gastric side-effects. PMID- 10383562 TI - Prediction of the international normalized ratio and maintenance dose during the initiation of warfarin therapy. AB - AIMS: A pharmacokinetic/pharmacodynamic model, with Bayesian parameter estimation, was used to retrospectively predict the daily International Normalized Ratios (INRs) and the maintenance doses during the initiation of warfarin therapy in 74 inpatients. METHODS: INRs and maintenance doses predicted by the model were compared with the actual INRs and the eventual maintenance dose. Cases with drugs or medical conditions interacting with warfarin or receiving concurrent heparin therapy were not excluded. As the study was retrospective, model predictions of the maintenance dose were not those that were administered. Mean prediction error (MPE) and percentage absolute prediction errors (PAPE) were used to assess the model predictions. RESULTS: INR MPE ranged from -0.07 to 0.06 and median PAPE from 10% to 20%. Dose MPE ranged from -0.7 to 0.17 mg and median PAPE from 16.7% to 37.5%. Accurate and precise dose predictions were obtained after 3 or more INR feedback's. CONCLUSIONS: This study shows that the model can accurately predict daily INRs and the maintenance dose in this sample of cases. The model can be incorporated into computer decision support systems for warfarin therapy and may lead to improvement in the initiation of warfarin therapy. PMID- 10383563 TI - Pharmacokinetic and pharmacodynamic characterization of OROS and immediate release amitriptyline. AB - AIMS: To characterize the pharmacokinetics of amitriptyline and its metabolite nortriptyline following OROS and IR treatments, and to correlate them with anticholinergic side-effects. METHODS: The pharmacokinetics and safety of amitriptyline following administration of an osmotic controlled release tablet (OROS and an immediate release (IR) tablet were evaluated in 14 healthy subjects. In this randomized, open label, three-way crossover feasibility study, the subjects received a single 75 mg OROS tablet, three 25 mg IR tablets administered every 8 h, or 3x25 mg IR tablets administered at nighttime. In each treatment arm serial blood samples were collected for a period of 84 h after dosing. The plasma samples were analysed by gas chromatography for amitriptyline and its metabolite nortriptyline. Anticholinergic effects such as saliva output, visual acuity, and subject-rated drowsiness and dry mouth were measured on a continuous scale during each treatment period. RESULTS: Following dosing with OROS (amitriptyline hydrochloride), the mean maximal plasma amitriptyline concentration Cmax (15.3 ng ml-1 ) was lower and the mean tmax (25.7 h) was longer than that associated with the equivalent IR dose administered at nighttime (26.8 ng ml-1 and 6.3 h, respectively). The bioavailability of amitriptyline following OROS dosing was 95% relative to IR every 8 h dosing, and 89% relative to IR nighttime dosing. The metabolite-to-drug ratios after the three treatment periods were similar, suggesting no change in metabolism between treatments. The relationships between plasma amitriptyline concentration and anticholinergic effects (e.g. reduced saliva weight, dry mouth, and drowsiness) were similar with all three treatments. Of the anticholinergic effects, only decreased saliva weight and dry mouth correlated well with plasma amitriptyline concentrations; drowsiness did not. There was no apparent correlation between anticholinergic effects and the plasma nortriptyline concentration. CONCLUSIONS: The bioavailability of OROS (amitriptyline hydrochloride) was similar to that of the IR treatments and the pharmacokinetics of amitriptyline after OROS dosing may decrease the incidence of anticholinergic effects compared with that seen with nighttime dosing of the IR formulation. Therefore, this controlled-release formulation of amitriptyline may be appropriate for single daily administration. PMID- 10383564 TI - Photoinduced covalent binding of frusemide and frusemide glucuronide to human serum albumin. AB - AIMS: To study reaction of photoactivated frusemide (F) and F glucuronide (Fgnd metabolite) with human serum albumin in order to find a clue to clarify a mechanism of phototoxic blisters from high frusemide dosage. METHODS: F was exposed to light in the presence of human serum albumin (HSA). HSA treated with this method (TR-HSA) was characterized by fluorescence spectroscopic experiment, alkali treatment and reversible binding experiment. RESULTS: Less 4-hydroxyl-N furfuryl-5-sulphamoylanthranilic acid (4HFSA, a photodegradation product of F) was formed in the presence of HSA than in the absence of HSA. A new fluorescence spectrum excited at 320 nm was observed for TR-HSA. Alkali treatment of TR-HSA released 4HFSA. Quenching of the fluorescence due to the lone tryptophan near the warfarin-binding site of HSA was observed in TR-HSA. The reversible binding of F or naproxen to the warfarin-binding site of TR-HSA was less than to that of native HSA. These results indicate the photoactivated F was covalently bound to the warfarin-binding site of HSA. The covalent binding of Fgnd, which is also reversibly bound to the warfarin-binding site of HSA, was also induced by exposure to sunlight. Fgnd was more photoactive than F, indicating that F could be activated by glucuronidation to become a more photoactive compound. CONCLUSIONS: The reactivity of photoactivated F and Fgnd to HSA and/or to other endogenous compounds may cause the phototoxic blisters that result at high F dosage. PMID- 10383566 TI - Comparison of the vasorelaxant effect of nitroprusside and nitroglycerin in the human radial artery in vitro. AB - AIMS: In recent years the radial artery (RA) has been re-introduced for coronary artery bypass grafting (CABG). However, the potential for vasospasm remains a clinical problem when this vessel is employed and effective vasodilator agents are required to combat vasospastic events. This in vitro study was designed to compare the vasodilator effects of sodium nitroprusside (SNP) and nitroglycerin (NTG) in the human RA. METHODS: Human RA segments (n=70) were taken from vessels employed for grafting in patients undergoing CABG. Concentration-relaxation curves for SNP and NTG were established in RA which had been precontracted with various vasoconstrictors (potassium chloride [K+], the thromboxane A2 mimetic agent U46619 or endothelin-1 [ET-1]). RESULTS: Both SNP and NTG caused complete relaxation and EC50s were similar except that NTG was 6.2-fold more potent than SNP in U46619-induced contraction (-7.50+/-0.16 vs -6. 71+/-0.38 log m, P=0.04). After treatment with verapamil and NTG solution during harvesting, the RA segments responded with reduced maximal relaxation to NTG (84.9+/-3.9%, compared with 98.8+/-0.8% in the control, P=0.004). The vessel became less sensitive to NTG (EC50: -6.29+/-0.4 vs -7.50+/-0.16 log m, P=0.01). In investigations carried out with SNP, tolerance was only seen in the magnitude of the relaxation (87.4+/ 4.7% vs 99.2+/-0.6% in the control, P=0.03). CONCLUSIONS: Both NTG and SNP are potent vasodilators in the RA. NTG may have more potent effects in certain situations (constriction related to thromboxane A2). However, tolerance to NTG may develop. A cross tolerance to SNP may exist but the effect is weak so that SNP may be preferable to NTG as a vasodilator in the RA postoperatively. Other vasodilators may be the drugs of choice under such circumstances. PMID- 10383565 TI - Zolpidem metabolism in vitro: responsible cytochromes, chemical inhibitors, and in vivo correlations. AB - AIMS: To determine the human cytochromes mediating biotransformation of the imidazopyridine hypnotic, zolpidem, and the clinical correlates of the findings. METHODS: Kinetic properties of zolpidem biotransformation to its three hydroxylated metabolites were studied in vitro using human liver microsomes and heterologously expressed individual human cytochromes. RESULTS: The metabolic product termed M-3 accounted for more than 80% of net intrinsic clearance by liver microsomes in vitro. Microsomes containing human cytochromes CYP1A2, 2C9, 2C19, 2D6, and 3 A4 expressed by cDNA-transfected human lymphoblastoid cells mediated zolpidem metabolism in vitro. The kinetic profile for zolpidem metabolite formation by each individual cytochrome was combined with estimated relative abundances based on immunological quantification, yielding projected contributions to net intrinsic clearance of: 61% for 3 A4, 22% for 2C9, 14% for 1A2, and less than 3% for 2D6 and 2C19. These values were consistent with inhibitory effects of ketoconazole and sulfaphenazole on zolpidem biotransformation by liver microsomes. Ketoconazole had a 50% inhibitory concentration (IC50 ) of 0.61 microm vs formation of the M-3 metabolite of zolpidem in vitro; in a clinical study, ketoconazole coadministration reduced zolpidem oral clearance by approximately 40%, somewhat less than anticipated based on the IC50 value and total plasma ketoconazole levels, but much more than predicted based on unbound plasma ketoconazole levels. CONCLUSIONS: The incomplete dependence of zolpidem clearance on CYP3A activity has clinical implications for susceptibility to metabolic inhibition. PMID- 10383567 TI - Trauma centres in the USA - past and present. PMID- 10383568 TI - Trauma centres: a British perspective. PMID- 10383569 TI - Clinical dilemma: A parathyroid adenoma cannot be found during neck exploration of a patient with presumed primary hyperparathyroidism. How should this problem be tackled? PMID- 10383570 TI - Multimodal therapy for squamous carcinoma of the oesophagus. AB - BACKGROUND: The results of surgical treatment for oesophageal squamous cell cancer have improved over recent decades, but the long-term prognosis for patients with tumours of stage II or higher is still unsatisfactory. While uncontrolled series of adjuvant or neoadjuvant treatment have reported favourable survival rates, the true benefit of multimodal treatment can be determined only by randomized controlled trials. METHODS: The literature was searched for prospective randomized controlled trials (PRCTs) examining the effect of neoadjuvant or adjuvant treatment on the long-term survival of patients with squamous cell cancer of the oesophagus. RESULTS: More than 30 PRCTs studying multimodal treatment concepts aimed at improving the prognosis of squamous cell carcinoma of the oesophagus were identified. These trials have not documented improved survival by adjuvant radiotherapy or chemotherapy. Following neoadjuvant radiotherapy or chemotherapy (or a combination of both), resectability of squamous cell carcinoma is not increased, the postoperative mortality rate appears to be higher and survival is not prolonged. CONCLUSION: Past multimodality protocols have not improved the prognosis of squamous carcinoma of the oesophagus. Careful design and reporting, together with strict control of surgical procedures, should allow more meaningful analysis of future multimodal treatment studies. At present, neoadjuvant or adjuvant treatment cannot be recommended outside such clinical protocols. PMID- 10383571 TI - Trefoil peptides and surgical disease. AB - BACKGROUND: Trefoil peptides are a family of small proteins that are expressed in a site-specific fashion by certain epithelial tissues. These peptides appear to be important in mucosal healing processes and in neoplastic disease. METHODS: This manuscript reviews the relevant literature obtained by an extensive text word search of the Medline database and a manual search of references from the articles identified. RESULTS AND CONCLUSION: Trefoil peptides are aberrantly expressed by a wide range of human carcinomas and gastrointestinal inflammatory conditions. They impart protection from injury to the gastrointestinal mucosa by possible interaction with mucin glycoproteins. Trefoil peptides influence epithelial cell migration and mucosal restitution following injury. In the future, serum levels of trefoil peptides might be used as markers for both neoplastic and inflammatory diseases. In addition, novel therapies based on such peptides might be used for gastrointestinal inflammatory conditions and to accelerate repair of the gastrointestinal mucosa after surgery. PMID- 10383573 TI - Digest PMID- 10383572 TI - Digest PMID- 10383574 TI - Role of superficial venous surgery in the treatment of venous ulceration. AB - BACKGROUND: The aim of this study was to determine the ability of superficial venous surgery to heal venous ulcers in lower legs with isolated superficial venous incompetence. METHODS: This was a prospective study of patients recruited from a venous ulcer assessment clinic. Ulcers were considered venous if the ankle : brachial pressure index was greater than 0.8 and duplex imaging showed venous reflux. Patients with isolated superficial venous incompetence were offered saphenofemoral and/or saphenopopliteal surgery. Neither perforator surgery, skin grafting nor postoperative compression hosiery or bandaging was used. RESULTS: A total of 122 legs with normal deep veins underwent superficial venous surgery. Ninety procedures (74 per cent) were done under local and 32 (26 per cent) under general anaesthesia. Sixty operations (49 per cent) were done as a day case. The median time to healing was 18 (95 per cent confidence interval 14-21) weeks and the cumulative 6-, 12- and 18-month healing rates were 57, 74 and 82 per cent respectively. CONCLUSION: In patients with venous ulceration and isolated superficial venous incompetence, superficial venous surgery can produce ulcer healing in the majority of patients without the need for perforator surgery, postoperative compression bandaging or skin grafting. PMID- 10383575 TI - Physiological comparison of open and endovascular aneurysm repair. AB - BACKGROUND: This prospective study compared morbidity and mortality rates following conventional and endovascular abdominal aortic aneurysm (AAA) repair using a physiological scoring system. METHODS: Between December 1994 and November 1997, 104 elective open aneurysm repairs and 49 endovascular aneurysm repairs were performed. These patient cohorts were compared using the Portsmouth predictor equation (P-POSSUM) scoring system. Data collected prospectively from patient notes were used to obtain physiological and operative severity scores which were analysed to compare expected and observed mortality and morbidity rates. RESULTS: There were three deaths (6 per cent) in the endovascular AAA repair group and 17 (16 per cent) in the conventional aneurysm repair group, whereas the P-POSSUM formulae predicted mortality rates of 8 and 19 per cent respectively. Although the mean physiological scores were similar for both groups (endovascular 20.8 versus conventional 20.1), the operative severity score was significantly greater in the conventional group (26.3 versus 19.7; P < 0.001). CONCLUSION: In this study open aortic aneurysm repair had a higher operative severity than endovascular repair, which was reflected in the increased mortality rate. PMID- 10383576 TI - Mortality from ruptured abdominal aortic aneurysm in Wales. AB - BACKGROUND: The aim of this study was to identify the incidence of, and mortality in, patients with a ruptured abdominal aortic aneurysm (AAA) reaching hospital alive in Wales. METHODS: Patients who presented with a ruptured AAA between September 1996 and August 1997 were analysed. Data were collected prospectively by an independent body, observing strict confidentiality. RESULTS: Some 233 patients with a confirmed ruptured AAA were identified, giving an incidence of eight per 100 000 total population. Some 133 patients (57 per cent) underwent attempted operative repair; 85 (64 per cent) of these died within 30 days. Of the 233 patients, 92 were admitted under the care of a vascular surgeon and 141 under a non-vascular surgeon. Vascular surgeons operated on 82 patients (89 per cent), of whom 50 (61 per cent) died, whereas non-vascular surgeons operated on 51 patients (36 per cent), of whom 35 (69 per cent) died. DISCUSSION: This study is unique as it is an independent prospective study of mortality in patients with a ruptured AAA who reached hospital alive. Mortality was independent of the operating surgeon, but vascular surgeons turned down significantly fewer patients than non-vascular surgeons (11 versus 64 per cent, P < 0.001). PMID- 10383577 TI - Uptake of tetracycline by aortic aneurysm wall and its effect on inflammation and proteolysis. AB - BACKGROUND: Proteolytic degradation of the aortic wall by matrix metalloproteinases (MMPs) is considered important in the pathogenesis of abdominal aortic aneurysms (AAAs). Many of these MMPs are inhibited by tetracycline derivatives, which may have the potential to retard aneurysm growth. METHODS: Patients undergoing elective repair of an AAA (n = 5) received an intravenous bolus of tetracycline (500 mg) on induction of anaesthesia and levels of tetracycline in serum, aneurysm wall and mural thrombus were assessed by microbiological assay. In a separate series of patients (n = 7) aneurysm biopsies were placed into explant culture (with and without tetracyline) and the accumulation of protein, hydroxyproline, MMP-9, interleukin (IL) 6 and monocyte chemoattractant protein (MCP) 1 in the medium was assessed by colorimetric assay or immunoassay. RESULTS: At aortic cross-clamping the median concentration of tetracycline was 8.3 microg/ml in serum, 2.9 microg per g tissue in aortic wall and zero in mural thrombus. Tetracycline inhibited, in a concentration-dependent manner, both MMP-9 and MCP-1 secretion (P = 0.022 and P = 0.018 respectively), but did not alter hydroxyproline or IL-6 secretion. At the highest concentration of tetracycline (100 microg/ml) median MMP-9 secretion was reduced from 27 to 5 ng/ml (P = 0.007) and median MCP-1 secretion was reduced from 50 to 10 ng/ml (P = 0.008). CONCLUSION: Tetracycline rapidly penetrates the aortic wall, but the concentration achieved may be insufficient to alter collagen turnover through limitation of MMP production or activity. PMID- 10383578 TI - Experimental study of the effect of intraportal prostaglandin E1 on hepatic blood flow during reperfusion after ischaemia and hepatectomy. AB - BACKGROUND: Prostaglandin E1 (PGE1) has protective effects experimentally and clinically in individual models of hepatic ischaemia-reperfusion injury and of partial hepatectomy. The present study investigated the effects of intraportal administration of PGE1 on hepatic blood flow, systemic arterial pressure and long term animal survival after 60 min of total liver ischaemia followed by 70 per cent partial hepatectomy in rats. METHODS: Total liver ischaemia was induced by occluding the hepatoduodenal ligament for 60 min. PGE1 0.5 microg per kg per min was infused intraportally for 15 min before inducing ischaemia and for 120 min after ischaemia in the treatment group. Normal saline was infused in the control group. During ischaemia 70 per cent partial hepatectomy was performed. Portal venous flow (PVF), peripheral tissue blood flow (PTBF) and hepatic artery flow were measured before and after ischaemia. Serum biochemical analysis was carried out at 1, 3 and 24 h, and 7 and 14 days; and liver histology at 1 and 24 h, and 7 days after reperfusion. Survival was followed for 1 year. RESULTS: Intraportal infusion of PGE1 significantly improved PVF and PTBF without affecting the systemic arterial pressure. Long-term survival was significantly higher in the PGE1 group. Serum aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase levels decreased significantly, and 2-h bile flow was significantly improved, in the PGE1 group. Histological examination revealed significant portal venous congestion, sinusoidal congestion, fatty degeneration and tissue necrosis 24 h and 7 days after reperfusion in the control group. CONCLUSION: PGE1 has a protective effect against liver damage when the liver is injured by warm ischaemia and reperfusion followed by partial resection. PMID- 10383579 TI - During liver regeneration following right portal embolization the growth rate of liver metastases is more rapid than that of the liver parenchyma. AB - BACKGROUND: The relative growth rates of human liver metastases are not known. The aim of this study was to determine in humans the growth rate of liver parenchyma during regeneration and the growth rate of liver metastases during the same process. METHODS: Among 556 patients hepatectomized for a malignant lesion, 48 underwent preoperative selective right portal vein embolization to induce hypertrophy of the left lobe. Five cases were selected because a liver metastasis was present inside the regenerating left lobe. The volumes of the liver metastasis and left lobe were measured with a three-dimensional technique on pre embolization and postembolization computed tomography (CT) or CT arterial portography. The median interval between the two measurements was 34 (range 28 40) days. RESULTS: An increase occurred in the volume of the liver metastasis and the left lobe in the four patients with functionally intact liver parenchyma, but not in the patient with an impaired parenchyma. The volumetric increase of the normal liver varied from 59 to 127 per cent, compared with 60 to 970 per cent for the liver metastases. The ratio between the growth rate of the left lobe and the liver metastasis varied from 1.0 to 15.6. The growth rates of two metastases in the left lobe of a single patient were different. CONCLUSION: In functionally intact liver parenchyma, during hepatic regeneration or hypertrophy, the growth rate of metastases is more rapid that that of the liver parenchyma. However, a wide variation in growth rate is observed between patients and between metastases. PMID- 10383580 TI - Spiral computed tomography for preoperative staging of potentially resectable carcinoma of the pancreatic head. AB - BACKGROUND: Pancreatic cancer is often locally invasive. Preoperative staging attempts to identify patients suitable for resection, in order to minimize unnecessary operations. The aim of this study was to assess the improved imaging provided by spiral computed tomography (CT) in the preoperative staging of potentially resectable pancreatic head carcinoma. METHODS: In 56 consecutive patients with pancreatic head carcinoma spiral CT findings were correlated prospectively with operative and histopathological findings. Criteria for irresectability at CT were infiltration of the peripancreatic fat and vascular ingrowth grade D, on a scale from A to F. RESULTS: At operation 27 (48 per cent) of 56 tumours were irresectable. Small metastases were found in seven patients (12 per cent). Ingrowth (adherence) to the portal or mesenteric vein was present in 19 patients (34 per cent). The sensitivity and specificity of CT for irresectability were 78 and 76 per cent respectively. Resection rates with a vascular margin free of tumour were 100 per cent for grade A, 63 per cent for grade B, 44 per cent for grade C, 15 per cent for grade D and 0 per cent for grade E, with a predictive value for ingrowth of 88 per cent for grades D or higher. The resectability rate was 11 per cent (one of nine) when infiltration of the anterior peripancreatic fat was present and 67 per cent when infiltration was absent (P < 0.01). CONCLUSION: Spiral CT with thin slices seems to improve detection of distant metastases and vascular ingrowth in patients with pancreatic head carcinoma. PMID- 10383581 TI - Endogenous renal nitric oxide metabolism following experimental infrarenal aortic cross-clamp-induced ischaemia-reperfusion injury. AB - BACKGROUND: Abdominal aortic surgery is associated with marked changes in renal haemodynamics. The aim of this study was to investigate the influence of infrarenal cross-clamping on glomerular filtration rate and endogenous renal nitric oxide metabolism. METHODS: Groups of male Wistar rats were subjected to infrarenal aortic cross-clamping followed by reperfusion. Animals were allowed to recover after a left nephrectomy. The glomerular filtration rate of the remaining kidney was measured on the second and seventh day after the procedure before the animal was killed and the remaining kidney harvested. Total nitric oxide synthase (NOS) activity and expression of inducible NOS (iNOS) was determined in renal tissue following 1 h and 7 days of reperfusion. RESULTS: Glomerular filtration rate was impaired on the second and seventh day after operation in all animals subjected to lower torso ischaemia compared with controls (P < 0.05). Renal NOS activity was increased at 1 h and 7 days in animals subjected to infrarenal cross clamping compared to controls (P < 0.01). iNOS was detected in renal tissue of animals subjected to infrarenal aortic cross-clamping on the seventh day after operation. CONCLUSION: Infrarenal aortic cross-clamping is associated with impairment of renal function in the early postoperative period. There is an increase in endogenous renal nitric oxide metabolism with iNOS expression. Presented in part to the Surgical Research Society, Dublin, Ireland, July 1998 PMID- 10383582 TI - Peripheral thrombolysis for acute-onset claudication. Thrombolysis Study Group. AB - BACKGROUND: The aim of this study was to determine the outcome of patients who presented with sudden onset of incapacitating claudication of less than 2 weeks' duration and who were treated with peripheral arterial thrombolysis. METHODS: The database of the Thrombolysis Study Group was searched retrospectively for patients who received thrombolysis for acute-onset claudication. Some 108 patients (65 men, median age 69 (range 29-94) years) were treated with intra arterial tissue plasminogen activator at 14 hospitals. The median duration of symptoms was 72 h (range from 2 h to 2 weeks). There were 52 graft and 56 native vessel arterial occlusions. RESULTS: The immediate outcome of thrombolysis for native vessel arterial occlusion was thrombus clearance in 50 patients (89 per cent) and failed lysis in six (11 per cent). Thirty-six patients (64 per cent) had a secondary radiological or surgical procedure carried out after lysis. After 30 days four patients (7 per cent) had a major amputation, eight (14 per cent) had died, 38 (68 per cent) were symptom free and seven (12 per cent) continued to have claudication. Three patients (5 per cent) suffered a major haemorrhage. The immediate outcome of thrombolysis for graft occlusion was thrombus clearance in 48 patients (92 per cent) and failed lysis in four (8 per cent); 27 patients (52 per cent) had a secondary procedure. After 30 days four patients (8 per cent) had a major amputation, seven (13 per cent) had died, 32 (62 per cent) were symptom free and nine (17 per cent) had persistent claudication. Three patients (6 per cent) suffered a major haemorrhage. CONCLUSION: Patients who presented with acute onset of incapacitating claudication had an outcome similar to that after thrombolysis for critical ischaemia. It is recommended that patients who present in this way should be observed and treated with thrombolysis only if they progress to critical ischaemia. Presented to the Association of Surgeons of Great Britain and Ireland, Edinburgh, UK, May 1998, and published in abstract form as Br J Surg 1998; 85(Suppl 1): 24 PMID- 10383583 TI - Phase II trial of radical surgery for locally advanced pelvic neoplasia. AB - BACKGROUND: Reported operative mortality and survival rates following total pelvic exenteration (TPE) for recurrent pelvic neoplasia are now as good as those for many primary treatments. The currently accepted primary treatments for these tumours are, however, still either radiotherapy alone or radiotherapy and chemotherapy. The primary aim of this study was to evaluate the safety and tolerability of TPE and secondarily to ascertain survival after TPE. METHODS: This was a phase II study of 50 patients with locally advanced pelvic tumours who underwent TPE. RESULTS: Thirty-two patients (64 per cent) underwent TPE for recurrent carcinoma of the cervix, seven (14 per cent) for rectal cancer, three (6 per cent) for vulval carcinoma, three (6 per cent) for vaginal carcinoma, two (4 per cent) for prostate cancer and three (6 per cent) for other tumours. The 30 day mortality rate was 8 per cent with an in-hospital mortality rate of 16 per cent. The crude morbidity rate was 62 per cent, with 23 patients (46 per cent) having grade III or IV toxicity. A complete response was achieved in 63 per cent and a partial response in 37 per cent of patients. The overall median survival time was 86 weeks; it was 111 weeks in patients in whom a complete response was achieved. CONCLUSION: The survival and operative mortality rates that are now attainable with TPE are comparable to those achieved with chemoradiotherapy in advanced pelvic neoplasia. TPE should no longer be reserved for salvage therapy and should perhaps be compared with chemoradiotherapy as first-line treatment in a phase III randomized trial in patients with these tumours. PMID- 10383584 TI - Selective impairment of glucose storage in human sepsis. AB - BACKGROUND: Glucose utilization in sepsis is impaired but the mechanisms are unclear. This study examined the effect of sepsis on total glucose utilization, oxidation and storage, and the energetic costs of these metabolic processes. METHODS: Glucose infusion rate (GIR), glucose oxidation rate (GOR), non-oxidative disposal rate and the energetic cost of glucose storage were studied in 24 patients with abdominal sepsis and in 26 healthy controls, using indirect calorimetry and the euglycaemic hyperinsulinaemic clamp with insulin infusion rates of 40 and 240 mU m-2 min-1. RESULTS: Basal GOR was significantly lower in septic patients than in controls (1.5 versus 2.3 mg per kg fat-free mass (FFM) per min, P < 0.001). Septic patients had a significantly lower GIR at 40 mU m-2 min-1 (4.2 versus 9.1 mg per kg FFM per min) and at 240 mU m-2 min-1 (7.5 versus 11.8 mg per kg FFM per min), relative to controls (P < 0.001). GOR was similar in septic and control subjects at both rates of insulin infusion whereas non oxidative disposal was significantly lower in septic patients (P < 0.001) and accounted entirely for the reduction in GIR. The energetic cost of glucose disposal was unaffected by sepsis. CONCLUSION: Sepsis is associated with selective impairment of glucose storage but the energetic cost of non-oxidative disposal is unaffected. PMID- 10383585 TI - Regulating a regulator: IFNgamma production by the neonate. PMID- 10383586 TI - The role of dust mite antigen sensitization and atopic dermatitis. PMID- 10383587 TI - Can eosinophil-derived proteins be used to diagnose or to monitor childhood asthma? PMID- 10383588 TI - Elements of the history of our present concepts of anaphylaxis, hay fever and asthma. PMID- 10383589 TI - The recombinant allergen-based concept of component-resolved diagnostics and immunotherapy (CRD and CRIT). PMID- 10383590 TI - On early sensitization to allergens and development of respiratory symptoms. AB - BACKGROUND: Various studies have suggested that a sequence of events occurring in childhood may affect the development of asthma in susceptible individuals. We have investigated whether early childhood sensitization to aeroallergens is an important risk factor in the later development of asthma symptoms. OBJECTIVE AND METHODS: In this study we examine this issue in children enrolled in the Tucson epidemiology study of obstructive airways disease, who had at least two allergen skin tests, one before and one after 8 years of age. Respiratory symptom data were available from 12 survey questionnaires, spanning a period of 20 years. During the first, sixth, seventh and eleventh surveys, skin tests were performed with commercially available allergens. CONCLUSION: As compared with children who were sensitized after 8 years of age, children over 8 years who were sensitized to any allergen before age 8 years were significantly more likely to report shortness of breath with wheeze (SOBWZ), wheeze apart from colds or wheeze most days (OR = 4.1 SOBWZ; OR = 3.88 WZ apart from colds; and OR = 2.83 WZ most days). Children who were sensitized after 8 years were no more likely to have the symptoms described above than children who were never found to be sensitized. Based on these results we conclude that early allergic sensitization is a significant risk factor for later development of wheezy symptoms, where as late sensitization is not. PMID- 10383591 TI - Mechanisms of deficient interferon-gamma production in atopic diseases. AB - BACKGROUND: The mechanisms responsible for an imbalanced cytokine response in atopic diseases are still not understood. While impaired interferon-gamma (IFN gamma) production may be the result of a pathological T-cell/antigen-presenting cell (APC) interaction, evidence was provided that the T cell itself may have an intrinsic defect to produce IFN-gamma. OBJECTIVE: To clarify whether impaired IFN gamma production by T cells from patients with atopic dermatitis (AD) represents an intrinsic defect in producing IFN-gamma. METHODS: Effector T cells were generated from CD4+ CD45RA+-naive precursors from patients with AD and healthy control individuals by activation with anti-CD3+ anti-CD28 MoAbs. Following restimulation, IFN-gamma production was measured by ELISA and flow cytometry. RESULTS: IFN-gamma production by atopic T cells was decreased compared with healthy T cells. IL-12 present at priming or high doses of IL-2 during the culture period, even in the absence of IL-12, completely restored IFN-gamma production. Conversion of naive CD45RA+ to CD45R0+ effector cells did not differ between atopic and healthy donors' T cells. CONCLUSION: Impaired IFN-gamma production by T cells from atopic individuals is not the result of an intrinsic, genetically fixed, defect to produce sufficient amounts of IFN-gamma. The data provides evidence that correction of an impaired TH1 response in AD may be successful at the precursor T cell level. PMID- 10383592 TI - Inverse correlation of domestic exposure to Dermatophagoides pteronyssinus antigen patch test reactivity in patients with atopic dermatitis. AB - BACKGROUND: In recent years considerable interest in the pathogenetic role of aeroallergens exacerbating atopic dermatitis (AD) has emerged. The 'atopy patch test' with aeroallergens was introduced by Platts-Mills et al. as an experimental model and as a diagnostic tool. However, its relevance for the clinical manifestation of AD is still not clear. OBJECTIVE: We asked whether there is a relationship between the individual antigen exposure to the major allergen of Dermatophagoides pteronyssinus (Der p 1) and the immunological markers of sensitization to Der p 1 or the clinical severity of AD. METHODS: We investigated 92 patients with moderate to severe AD. For clinical evaluation the SCORAD severity score was used. Patch tests were performed with purified Der p 1. Specific IgE was measured by a commercial assay. Der p 1 exposure was quantified in a sample of the patient's mattress dust by using a commercial ELISA. RESULTS: No correlation between SCORAD, Der p 1 exposure and RAST could be established. However, there was an unexpected significant inverse correlation between the quantity of mite antigen in the mattress dust and patch test reactivity. Patients with a high antigen load (> 25 microg/g) mostly had a negative patch test. Also, when Der p 1 was correlated to the mattress area (m2) in this group all patch tests were negative. A possible explanation could be that continuous exposure of the skin to house dust mite allergen Der p 1 may induce a down-regulation of the skin immune system of patients with AD. CONCLUSION: Although the mechanism of this phenomenon is presently unknown, our study shows that a positive allergen patch test alone should not be an indication to undertake allergen exclusion measures in AD patients. PMID- 10383593 TI - Lack of relationship between eosinophil cationic protein and eosinophil protein X in nasal lavage and urine and the severity of childhood asthma in a 6-month follow-up study. AB - BACKGROUND: Recent studies suggest that eosinophil cationic protein (ECP) and eosinophil protein X (EPX) may be valuable markers of airway inflammation in various body fluids of asthmatic children. Most of these studies have relied on a single measure of inflammatory markers. OBJECTIVE: We measured ECP and EPX in nasal lavage fluids (NALF) and urine samples in children with asthma over a 6 month period to study the relationship between inflammatory markers and clinical severity. METHODS: Fourteen children with mild persisting asthma (mean age 11.7 years, SD 2.2) were recruited. All patients were on therapy including inhaled steroids. For a 6-month period asthma severity was monitored by at least monthly physical examination and pulmonary function tests. Daily morning and evening PEF, asthma symptoms and medication were recorded in diaries for the whole study period. Telephone interviews were performed between visits and additional visits were done in case of an increase in asthmatic symptoms or drop of PEF values under 80% of best value. An exacerbation was defined by a fall of FEV1 > 10% and an increase in asthma symptoms and additional need of beta2-agonist. NALF and urine samples were obtained at each visit and analysed for ECP (NALF only) and EPX. RESULTS: Mean observation time was 186.4 days (SD 19.8). Thirteen patients completed the study. During the study period 11 exacerbations were observed in six patients. No significant associations between PEF, PEF variability (amplitude % of mean), daily symptoms, additional beta2-agonist, FEV1 and MEF50 and nasal ECP, nasal EPX and urinary EPX were found. However, at exacerbations an average increase of nasal ECP (9.3 vs 50.3 microg/L) and EPX (nasal EPX 36.4 vs 141.7 microg/L, urinary EPX 46.4 vs 74.1 microg/mmol creatinine) was observed. CONCLUSION: Serial measurements of ECP and EPX in NALF and urine samples do not provide additional information for the practical management in monitoring childhood asthma. PMID- 10383594 TI - Allergen challenge primes for IL-5 mRNA production and abrogates beta-adrenergic function in peripheral blood T lymphocytes from asthmatics. AB - BACKGROUND: In previous studies, we have found a dysfunctional adenylyl cyclase (AC) system in patients with asthma after allergen provocation, which resulted in a 40-50% decreased generation of intracellular cAMP. In addition, in activated T helper lymphocyte clones, it has been demonstrated that IFN-gamma (TH1-like cytokine) and IL-5 (TH2-like cytokine) are differentially regulated by the AC system. Therefore, we postulate that an increased IL-5/IFN-gamma ratio as observed in asthmatics might be due to a dysfunctional AC system. OBJECTIVE: To assess whether a dysfunctional AC system as observed in asthmatics after allergen provocation, is responsible for an increased IL-5/IFN-gamma cytokine ratio. METHODS: Peripheral blood T lymphocytes of seven asthma patients were stimulated with anti-CD3 plus anti-CD28 MoAbs in the absence and presence of isoproterenol (ISO) and prostaglandin E2 (PGE2) to activate the AC system. Before, 3 h and 24 h after allergen provocation, IFN-gamma and IL-5 mRNAs were detected by semiquantitative RT-PCR. RESULTS: Before allergen provocation, ISO (10-5 mol/L) significantly downregulated IFN-gamma mRNA (P < 0.03, n = 6), and showed a trend to upregulate IL-5 mRNA (P = 0.138, n = 5). Three and 24 h after allergen provocation, ISO was not longer able to modulate IFN-gamma and IL-5. In contrast with the observations with ISO, PGE2 still dose-dependently inhibited IFN-gamma mRNA, both before, 3 h and 24 h after allergen provocation (n = 7). IL-5 mRNA, but not IFN-gamma mRNA, was significantly upregulated in anti-CD3 plus anti-CD28 activated T cells (P < 0.05, n = 5) 24 h after allergen provocation, compared with before allergen provocation. CONCLUSION: Twenty-four hours after allergen provocation, a significant reduction of beta-adrenergic control on IFN-gamma and IL-5 mRNA expression was observed in peripheral blood T lymphocytes, which coincides with a selective priming of IL-5 mRNA production. PMID- 10383595 TI - Nasal response to a single antigen challenge in patients with allergic rhinitis - inflammatory cell recruitment persists up to 48 hours. AB - BACKGROUND: Allergen challenge in some patients with respiratory allergy is followed by an early and a late reaction. OBJECTIVE: To evaluate the duration of mediator release and inflammatory cell recruitment during the late antigen induced nasal response. METHODS: Eight patients with seasonal allergic rhinitis due to grass pollen underwent local challenge with the relevant allergen, a non relevant allergen (Parietaria judaica), and nebulized saline solution. Nasal lavages were performed at baseline and 6, 24, 48, 72 h after challenge. Eosinophil cationic protein (ECP), leukotriene C4 (LTC4), leukotriene B4 (LTB4) myeloperoxidase (MPO) and prostaglandin D2 (PGD2) levels were radioimmunoassayed and histamine concentration was measured by an automated fluorometric method. RESULTS: Nasal challenge with the relevant antigen induced a response 6 h after stimulation, which subsided within 24 h. Eosinophilia, observed in the nasal lavages collected from 6 to 24 h after this challenge, was accompanied by ECP release. Neutrophilia were found in the nasal lavages collected from 6 to 24 h after challenge. The increase in neutrophil number correlated with MPO levels and LTB4 concentrations, but not with the intensity of nasal obstruction. Antigen challenge also induced significant recruitment of mononuclear cells 48 h after provocation. The challenge significantly raised histamine, but not PGD2, levels in the nasal lavages collected 6 h after provocation. A trend towards an increase in LTC4 levels in the nasal lavages collected 6 h after specific antigen challenge was also found. Nasal challenge with a non-relevant allergen or with saline solution did not cause either inflammatory cell recruitment or mediator release. CONCLUSION: Nasal challenge with the relevant antigen can induce a late response characterized by local accumulation of eosinophils, neutrophils and mononuclear cells persisting for 48 h and accompanied by release of ECP, MPO, LTB4 and histamine. These results indicate that a single antigen challenge in patients with allergic rhinitis causes prolonged inflammatory alterations which may contribute to the development of airway hyperreactivity. PMID- 10383596 TI - Anti-inflammatory effect of roxithromycin in patients with aspirin-intolerant asthma. AB - BACKGROUND: Fourteen-membered macrolides, such as roxithromycin, have been reported to exhibit other pharmacological activity including anti-asthmatic effects, besides antibiotic activity. OBJECTIVE: This study was designed to investigate the protective effect of roxithromycin on airway responsiveness to the sulpyrine provocation test and to investigate whether this protective activity is associated with a reduction in aspirin-induced excretion of urinary leucotriene E4 (u-LTE4), a marker of cysteinyl leucotriene overproduction that participates in the pathogenesis of aspirin-intolerant asthma. Also, the present study was designed to examine whether or not its anti-asthmatic activity was associated with a reduction in eosinophilic inflammation. METHODS: For 8 weeks before analysis, subjects received 150 mg of roxithromycin or matching placebo twice daily. We assessed the effects of pretreatment with roxithromycin on bronchoconstriction precipitated by inhalation of sulpyrine in 14 adult patients with mild or moderate aspirin-intolerant asthma; those who were in stable clinical condition and were hyperresponsive to sulpyrine provocation test were allocated to this study. A double-blind, randomized, crossover design was used. Urinary LTE4 was measured by a combined reverse-phase high-performance liquid chromatography (rp-HPLC) enzyme immunoassay on sulpyrine provocation testing day. Blood and sputum samples were taken in the morning on the sulpyrine provocation testing day. Eosinophil counting and measurement of eosinophilic cationic protein (ECP) were performed. RESULTS: After the 8 weeks of treatment with roxithromycin, patients' symptoms, blood eosinophils, serum ECP, sputum eosinophils, and sputum ECP were significantly decreased. On the other hand, values of PC20-sulpyrine did not improve after roxithromycin at all. Furthermore, although challenge with sulpyrine caused a significant increase in u-LTE4, pretreatment with roxithromycin or placebo did not affect excretion of u-LTE4. CONCLUSION: Although roxithromycin does not have antileucotriene effects, it has an antibronchial inflammatory effect associated with eosinophilic infiltration. This study raises further interesting therapeutic possibilities and warrants further trials of new approaches to the treatment of aspirin-intolerant asthma. PMID- 10383597 TI - Mucosal immunity in extrinsic allergic alveolitis: salivary immunoglobulins and antibody against inhaled avian antigens among pigeon breeders. AB - BACKGROUND: Inhaled antigens from pigeons can cause extrinsic allergic alveolitis (EAA); a model disease of pulmonary inflammation. Among pigeon breeders, serum antibody and sensitized lymphocytes specific for these antigens have been described primarily, but not always, with disease. Antibody activity within the lung may have a closer association with disease, however, sampling by alveolar lavage at bronchoscopy is impractical for screening, therefore we used saliva to quantify the mucosal antibody response. OBJECTIVE: To establish: (a) if antibody activity against inhaled avian antigens was detectable in the saliva of pigeon breeders, (b) if the distribution of saliva antibody and total immunoglobulin levels were quantitatively or qualitatively different from serum, and (c) whether the hypersensitivity symptoms of EAA were associated more with the mucosal or the systemic humoral immune response. MEASURES: Saliva and serum total and avian antigen-specific IgG, IgA (IgA1 and IgA2) antibody activity in 87 pigeon breeders and 24 control subjects with no avian exposure. Albumin levels were used as a protein reference and cotinine levels confirmed smoking status. Specific hypersensitivity symptoms and various exposure indices to pigeons were established by interview. RESULTS: Absolute levels and relative proportions (vs albumin) of IgG, IgA and IgA1 in saliva, and IgG in serum, were significantly higher in pigeon breeders compared with controls, suggesting mucosal inflammation. Avian antigen-specific antibody of all isotypes was readily demonstrable in saliva (predominantly IgA) and serum (predominantly IgG) from pigeon breeders, and there were no significant titres in controls. The levels of IgG antibody in saliva and in serum correlated significantly (r = 0.52, P < 0.001), and both correlated with the raised immunoglobulin levels. In both saliva and serum the IgG rather than the IgA antibody activity was associated with symptoms of EAA. CONCLUSIONS: Antibody activity in saliva and serum, representing the mucosal and systemic responses, respectively, were both strongly stimulated by inhaled antigens. The IgG antibody titres of saliva and serum correlated significantly and were a useful index of inflammation, as measured by the raised total immunoglobulin levels, and symptoms. This suggests that IgG antibody in serum may reflect clinical and immunological sensitization of the lung mucosa. Collecting saliva is noninvasive, and saliva antibody measurement is a convenient method for monitoring EAA, especially in children, and will facilitate sampling for example in epidemiological studies of antibody prevalence. PMID- 10383598 TI - Immunocytochemical localization of cyclo-oxygenase isoforms in cultured human airway structural cells. AB - BACKGROUND: Cyclo-oxygenase (COX) exists as two isoforms, COX-1, the constitutive isoform, and COX-2, which is inducible by cytokines or inflammatory stimuli and may participate in airway inflammation. OBJECTIVE: To determine the basal distribution of COX isoforms, and their regulation by interleukin-1 beta (IL 1beta), bradykinin (BK) and dexamethasone (Dex) in cultured airway structural cells. METHODS: We measured COX-1 and COX-2 in cultured human airway smooth muscle (HASM) cells, MRC5 fibroblasts and normal human epithelial cells (NHBE) using immunocytochemical analysis. RESULTS: The majority of all types of untreated cultured cells expressed COX-1 (75% of HASM, 75% of MRC5 fibroblasts and 72% of NHBE cells). Fibroblasts and smooth muscle cells showed low constitutive COX-2 expression (2 and 8%, respectively) but this was higher in NHBE cells (28%). IL-1beta (24 h incubation) or BK (4 h incubation) had no effect on COX-1 expression in any of the cells studied. In contrast, there was a two- and 1.5-fold rise in the percentage of NHBE cells expressing COX-2; a 7.5- and sixfold rise in the percentage of HASM cells expressing COX-2 and a 33.5- and 20.5-fold increase in the percentage of fibroblasts expressing COX-2 after IL 1beta or BK treatment, respectively. Pretreatment with dexamethasone abolished IL 1beta- and BK-stimulated COX-2 induction in all cells studied. CONCLUSION: COX-1 is expressed constitutively in human airway fibroblasts, smooth muscle and epithelial cells but epithelial cells also show constitutive expression of COX-2. Both IL-1beta and BK induced COX-2 expression in all cells studied and this induction was blocked by dexamethasone. Immunocytochemical techniques can be successfully used to detect the distribution of COX isoforms in cell cultures. PMID- 10383599 TI - Subtribe-specific monoclonal antibodies to Lolium perenne. AB - BACKGROUND AND OBJECTIVES: The ability to measure personal exposure to airborne grass pollen is important in the understanding of allergic diseases. Visual identification is time consuming and it is difficult to distinguish between many grass pollens morphologically. Although grass pollens share common allergenic determinants, we attempted to produce monoclonal antibodies that would distinguish between species, tribes and subfamilies of grasses which would allow immunodetection of pollens. METHODS: Monoclonal antibodies raised against Lolium perenne were screened for specificity against an extended panel of grass pollen extracts using standard ELISA techniques and a novel particle blotting assay using whole pollen grains. RESULTS: Antibodies showing specificity ranging from subfamily to part-tribe specificity were raised. The most specific monoclonal antibodies (numbers 4, 13 and 17) had reactivity to Lolium perenne and Festuca elatior but displayed little cross-reactivity to Phalaris arundinaceae and the rest of the Poeae tribe when tested by ELISA and no detectable cross-reaction when tested with particle blotting. CONCLUSION: Monoclonal antibodies that are functionally specific to only two grasses can be produced and used to discriminate between related grass species. PMID- 10383600 TI - Allergenic differences between the domestic mites Blomia tropicalis and Dermatophagoides pteronyssinus. AB - BACKGROUND: House dust mite allergens are the most important indoor allergens associated with asthma and rhinitis in Singapore and the tropics. Recent data to suggest that besides the Dermatophagoides spp., the domestic mite Blomia tropicalis (Bt) is also an important source of allergens in these regions. OBJECTIVE: To evaluate the degree of allergenic cross-reactivity between Bt and D. pteronyssinus (Dp). METHODS: Cross-reactivity between extracts of Bt and Dp was evaluated by fluorescent allergosorbent (FAST) inhibition studies and cross enzyme immunoelectrophoresis. Additionally, the major Dp allergens - Der p 1, Der p 2 and Der p 5, were also compared with the Bt extract by dot blot inhibition. Skin prick and intradermal end-point titration were then carried out to compare the homologous allergens of the mite species, Blo t 5 and Der p 5. RESULTS: FAST inhibition studies showed low to moderate cross-reactivity between the two dust mite extracts (maximum cross-inhibition, 60%). Native allergens studied by cross enzyme immunoelectrophoresis using mite allergic sera also showed similar results but with at least four cross-reactive IgE binding antigens. Dot blot inhibition studies using allergens of Dp, Der p 1, Der p 2, and Der p 5, showed little cross reactivity between these allergens with components of the crude Bt extracts. Further, evaluation of a recombinant allergen of Bt, Blo t 5, showed low levels of cross-reactivity even with its homologous Dp counterpart, Der p 5. CONCLUSION: These results provide evidence that Bt allergens are distinct and have relatively low to moderate cross-reactivity with Dermatophagoides spp. allergens. Bt allergens should therefore be included in the diagnostic panel for the evaluation of allergic disorders in the tropics, and the development of new diagnostic and therapeutic strategies should include allergens of Bt. PMID- 10383601 TI - Mutant derivatives of the main respiratory allergen of cow are less allergenic than the intact molecule. AB - BACKGROUND: Allergen immunotherapy offers an alternative for drug treatment in the management of allergic diseases. Because immunotherapy often induces side effects, less allergenic preparations would be beneficial. OBJECTIVE: The purpose of this study was to examine whether the allergenicity of a cow-derived lipocalin allergen, Bos d 2, could be diminished by substituting or deleting carboxy terminal amino acids including the cysteine which forms a disulphide bond with a cysteine inside the molecule. METHODS: Four recombinant mutants of Bos d 2 were created by substituting or deleting the four most carboxy-terminal amino acids. The immunological characteristics of the mutant preparations were compared with the unmodified rBos d 2 by Western blotting, ELISA inhibition, skin prick tests, and the proliferative responses of allergen-specific T-cell clones. RESULTS: In Western blot, one of the two monoclonal antibodies showed reduced binding to the preparations without the terminal cysteine. In contrast, the other monoclonal antibody, human IgE and rabbit immune serum bound equally well to all the preparations. ELISA inhibition analyses revealed, however, that the preparations without the terminal cysteine bound antibody less efficiently. They were needed 15-38 times more than the unmodified rBos d 2 to cause the same level of inhibition. Surprisingly, one of the mutants with the terminal cysteine but a mutated adjacent amino acid turned out to be the weakest in inducing skin reactivity. All the preparations stimulated well allergen-specific T-cell clones. CONCLUSIONS: The results show that the allergenicity of a lipocalin allergen, Bos d 2, can be diminished by modifying the carboxy-terminal end of the molecule. Modifications in the area which encompasses a disulphide bond impaired the antibody binding without affecting the T-cell stimulatory capacity. It was also shown that in vivo tests are necessary for determining the allergenicity of a modified allergen. PMID- 10383603 TI - Glucose metabolism and the postprandial state. AB - Disturbances of postprandial glucose metabolism are now thought to contribute to cardiovascular disease. Postprandial glucose excursions can be affected by a number of factors, such as the types of carbohydrates ingested and the way they are metabolized. In Type 2 diabetes, factors that contribute to excessive postprandial glucose excursions include disruption of insulin secretion, insufficient inhibition of hepatic glucose production and defective glucose storage in muscle. A number of measures may attenuate excessive postprandial blood glucose excursions. These include a diet high in 'low glycaemic index' foods and treatment with drugs that improve or restore the hormonal response (e.g. the sulphonylureas and the newer beta-cell mediated insulinotropic drugs such as repaglinide), that improve insulin sensitivity or that delay gastric emptying. PMID- 10383602 TI - Cross-reactivity between milk proteins from different animal species. AB - BACKGROUND: Cow's milk allergy is quite frequent in the first years of human life. When breast-feeding is not possible, a cow's milk substitute must be provided for allergic subjects. Different alternatives to cow's milk have been suggested as protein sources (soy, hydrolysed proteins, goat's milk, etc.), but all these dietetic solutions are not without risks for polyallergic or more sensitive subjects. OBJECTIVE: To obtain new information on the suitability of other mammalian milks for allergic children, we evaluated the cross-reactivity between milk proteins from different animal species. METHODS: Milk samples were analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS PAGE). To detect antibody-antigen complexes, immunoblotting was performed by using sera from children allergic to cow's and ewe's milk (RAST class >/= 4) and monoclonal antibodies (MoAb) specific for bovine proteins (caseins and beta lactoglobulin). RESULTS: IgEs from children allergic to cow's milk are capable of recognizing most part of milk proteins from mammals bred in European countries (ewe, goat, buffalo), while no serum used in this study contains IgEs reacting with camel's milk proteins. Camel's milk was also not recognized from circulating IgEs from a child specifically allergic to ewe's milk. Specific antibovine monoclonal antibodies cross-reacted with proteins from other mammalian species, apart from those of camel. CONCLUSIONS: Homologies in amino acidic composition could justify the cross-reactivity observed between proteins from different animal species. On the other hand, the phylogenetic difference could be responsible for the failed recognition of camel's proteins by circulating IgEs and monoclonal antibodies. PMID- 10383604 TI - Prandial glucose regulation and cardiovascular disease in type 2 diabetes. AB - Type 2 diabetes is associated with an increased risk for cardiovascular disease. In recent years, prospective studies have indicated that, in addition to conventional risk factors, glycaemic control of diabetes predicts cardiovascular disease in both middle-aged and elderly patients with Type 2 diabetes. However, there are no consistent data from different studies to indicate that postprandial glucose is a better predictor for cardiovascular risk than fasting glucose level. Although no clinical trials are available to show that improving glycaemic control prevents cardiovascular mortality and morbidity, recent studies imply that hyperglycaemia in patients with Type 2 diabetes should be treated more intensively than recommended by current guidelines PMID- 10383605 TI - Diabetic dyslipidaemia. AB - Type 2 diabetic patients have an increased risk of cardiovascular disease and, although many factors contribute to this risk, it is likely that diabetic dyslipidaemia plays an important role. Dyslipidaemia in Type 2 diabetic patients is characterized by low levels of HDL cholesterol and high triglyceride levels. In Type 2 diabetes, the total amount of LDL cholesterol is the same as in healthy people, but there are qualitative changes, e.g. a shift to smaller, denser LDL particles and an increased susceptibility to oxidation. Oxidized LDL may promote the development of atherosclerosis. It is possible to modify the major abnormalities of diabetic dyslipidaemia by combining lifestyle modifications (e.g. increased physical activity, cessation of smoking and weight reduction) with improved glycaemic control and hypolipidaemic drugs to reduce the burden of CVD within this high-risk population. PMID- 10383606 TI - Current therapeutic options in type 2 diabetes. AB - The prevalence of Type 2 diabetes rises steeply with age and involves beta-cell dysfunction and diminished sensitivity to insulin. beta-cell dysfunction is important in the development of hyperglycaemia while insulin resistance seems to play a major role in the atherogenic process resulting in cardiovascular disease. Current therapeutic options include lifestyle adjustments (exercise and diet), oral hypoglycaemic agents (sulphonylureas, newer beta-cell mediated insulin releasing drugs, alpha-glucosidase inhibitors, biguanides and thiazolidinediones) and insulin treatment. Oral hypoglycaemic agents are effective only temporarily in maintaining good glycaemic control, their efficacy should be determined from changes in fasting and postprandial glucose levels. Recent studies have shown that the early initiation of insulin therapy can establish good glycaemic control. PMID- 10383607 TI - Repaglinide, a new oral antidiabetic agent: a review of recent preclinical studies. AB - Repaglinide is a new carbamoylmethyl benzoic acid derivative that is structurally related to meglitinide. In common with all active oral hypoglycaemic agents, it displays a comparable U-shaped configuration. Repaglinide exerts its effects by binding to a site on the plasma membrane of beta-cells, thereby closing ATP sensitive potassium channels. This causes depolarization of the beta-cell and the opening of voltage-sensitive calcium channels allowing the influx of extracellular calcium ions. This increase in intracellular calcium, in turn, stimulates insulin release. The insulinotropic effect of repaglinide is not related to its ionophoretic capacity. Repaglinide does not cause insulin release in the absence of exogenous glucose, nor does it inhibit the biosynthesis of proinsulin. In vivo administration of an ester of succinic acid potentiates the insulin secretory response to concomitantly administered repaglinide. Repaglinide causes a more rapid increase in plasma insulin levels in rats than either glibenclamide or glimepiride. PMID- 10383608 TI - Repaglinide: a new short-acting insulinotropic agent for the treatment of type 2 diabetes. AB - Repaglinide, a carbamoylmethyl benzoic acid derivative, is rapidly absorbed, metabolized by the liver and eliminated primarily via the bile. It has a short duration of action and is taken immediately before each main meal. This regimen has been shown to provide superior glycaemic control compared with regular morning and evening dosing. A flexible preprandial only dosing regimen of repaglinide significantly lowers the risk of hypoglycaemia if a meal is missed or postponed. Combination therapy with metformin improves glycaemic control significantly compared with therapy with either drug alone in overweight patients. Repaglinide has an equivalent safety and efficacy profile to the sulphonylureas, although it is superior to glipizide in maintaining long-term glycaemic control The postprandial glucose levels are significantly lower with repaglinide compared with glibenclamide. In naive patients with Type 2 diabetes, repaglinide lowers fasting glucose concentrations and functions also as a prandial glucose regulator. PMID- 10383609 TI - C-fos and c-jun in the paraventricular nucleus play a role in regulating peptide gene expression, oxytocin and glutamate release, and maternal behaviour. AB - In sheep, birth leads to the induction of maternal behaviour through brain oxytocin release. Associated with these events is an upregulation of oxytocin, opioid and corticotrophin-releasing hormone (CRH) gene expression, as well as that of the immediate early gene c-fos in the paraventricular nucleus (PVN) of the hypothalamus. We investigated the role of c-fos dimerizing with c-jun in controlling the induction of maternal behaviour, altered peptide gene expression, and oxytocin and amino acid release in this region at birth. Fluorescence labelled antisense oligodeoxyribonucleotides (ODNs) against c-fos/c-jun were infused bilaterally in the PVN, via microdialysis probes with 100 kDa cut-off membranes, and were incorporated into 50-60% of the cells. Compared with the control (scrambled) sequences, they significantly reduced basal concentration of glutamate (to 31.7% of baseline after 10 h) and prevented birth-induced release of aspartate. In addition, antisense treatment reduced the birth-induced increase in oxytocin concentration in the PVN, but not in blood. Although all the animals were fully maternal, the antisense treatment did reduce the peak expression of two components of maternal behaviour: low-pitched bleats; and lamb sniffing. Finally, in situ hybridization histochemistry revealed that the antisense treatment significantly reduced the birth-induced upregulation of c-fos, oxytocin, CRH and preproenkephalin mRNA expression in the PVN, whilst not affecting that of arginine vasopressin. These results suggest that c-fos/c-jun transcription factors play a role in the birth-induced upregulation of oxytocin, CRH and preproenkephalin gene expression, as well as on glutamate and oxytocin release in the sheep PVN. PMID- 10383610 TI - The neuropeptide Y Y1 receptor is a somatic receptor on dorsal root ganglion neurons and a postsynaptic receptor on somatostatin dorsal horn neurons. AB - Using indirect immunofluorescence, neuropeptide Y Y1 receptor (Y1 receptor)-like immunoreactivity (LI) was localized close to the plasmalemma of small neurons in lumbar dorsal root ganglia (DRGs) and neurons in the inner lamina II of the lumbar spinal cord of the rat. Using confocal microscopy, colocalization of Y1 receptor-LI and transferrin receptor-LI, a marker for endosomes and coated vesicles, was observed in dot-like structures along the plasmalemma. Under the electron microscope, Y1 receptor-LI was localized in coated vesicles and endosomes, in the membrane of tubular cisternae, sometimes connected to multivesicular bodies, and in the plasmalemma. These complex distribution patterns may reflect receptor turnover and internalization processes. In the lamina II of the spinal dorsal horn, Y1 receptor-LI was localized in the plasmalemma of neurons without any apparent association with paramembrane structures, as described above for the DRG neurons. Many dendrites were Y1 receptor-positive, and some of them made synaptic contacts with unstained axonal terminals. In general, Y1 receptor-LI was localized in the membrane outside the postsynaptic density. Double-immunofluorescence staining showed that most Y1 receptor-immunoreactive neurons in lamina II contained somatostatin-LI. Both in DRG and dorsal horn neurons, the Y1 receptor thus seems to represent a postjunctional/postsynaptic receptor. PMID- 10383611 TI - Expression of glutamate transporters in rat optic nerve oligodendrocytes. AB - To investigate the role of glutamate transport in non-synaptic glia, we characterized the expression of three major glutamate transporters (EAAC1, GLAST and GLT-1) in rat optic nerve in situ using reverse transcription-polymerase chain reaction in combination with Western blot and immunochemistry with specific antibodies. GLAST was localized to interfascicular oligodendrocytes, whereas a subpopulation of cells, probably immature oligodendrocyte cells, expressed EAAC1. In contrast, astrocytes, expressed only GLT-1, consistent with the idea that this is the major glutamate transporter in this cell type. In addition, we observed that glutamine synthetase, a key enzyme in glutamate metabolism, was localized in oligodendrocytes in situ. To examine the properties of these glutamate transporters, we conducted uptake experiments in glial cultures. The kinetics of sodium-dependent glutamate uptake in cultured oligodendrocytes from the perinatal rat optic nerve were markedly different from those observed in type-1 astrocytes from the newborn rat cerebral cortex, with higher affinity and lower Vmax. In both cell types, glutamate transport was inhibited by L-trans-pyrrolidine-2,4 dicarboxylate (t-PDC). In contrast, dihydrokainate exhibited significantly more uptake inhibition in oligodendrocytes than in type-1 astrocytes. These results provide evidence for the expression of functional sodium-dependent glutamate transporters in optic nerve oligodendrocytes, and suggest that this cell type may play a role in the glutamate-glutamine cycle. PMID- 10383612 TI - Overexpression of GAP-43 induces prolonged sprouting and causes death of adult motoneurons. AB - In neurodegenerative diseases, neurons undergo prolonged periods of sprouting. Whether this sprouting compromises these neurons is unknown. Here, we examined the effect of axotomy on adult motoneurons undergoing prolonged sprouting in transgenic mice that overexpress GAP-43 (growth-associated protein). Sciatic nerve injury in these adult mice results in motoneuron death, but has no effect in non-transgenic mice. Thus, continued growth of motor axons renders adult motoneurons susceptible to nerve injury and compromises their long-term survival. The progressive nature of neurodegenerative diseases may therefore be caused by prolonged sprouting. PMID- 10383613 TI - Endogenous interleukin-6 contributes to hypersensitivity to cutaneous stimuli and changes in neuropeptides associated with chronic nerve constriction in mice. AB - Partial nerve injury is a potential cause of distressing chronic pain for which conventional analgesic treatment with opiates or anti-inflammatory agents is not very effective. Constriction nerve injury, widely used to study neuropathic pain, was shown here to induce interleukin-6 (IL-6) mRNA in a subset of rat primary sensory neurons. When we inflicted chronic nerve constriction on mice with null mutation of the IL-6 gene, the hypersensitivity to cutaneous heat and pressure that is induced in wild-type mice was not evident, the loss of substance P in sensory neurons was excessive and the induction of galanin in central sensory projections was reduced. In additional experiments, intrathecal infusion of IL-6 in rats was shown to stimulate synthesis of galanin in approximately one-third of lumbar dorsal root ganglion neurons. The results of these experiments indicate that endogenous IL-6 mediates some of the hypersensitive responses that characterize peripheral neuropathic pain, and influences two neuropeptides that have been implicated in pain transmission. PMID- 10383614 TI - Regionalization of Fos immunostaining in rat accessory olfactory bulb when the vomeronasal organ was exposed to urine. AB - The distribution of Fos-immunoreactive (Fos-ir) cells in the accessory olfactory bulb (AOB) of rats following vomeronasal organ exposure to urine was studied. Following exposure to male and female Wistar rat urine, Fos-ir cells were found in the mitral/tufted cell layer, granule cell layer and periglomerular cell layer of the AOB of female Wistar rat, with the highest number in the granule cell layer. Exposure to water or removal of the vomeronasal organ suppressed the expression of Fos-ir cells. These results suggest that female Wistar rats specifically detect urinary substances derived from male or female Wistar rats via the vomeronasal organ. Exposure of the vomeronasal organ of female Wistar rats to male Wistar urine induced the appearance of many more Fos-ir cells in all layers of the AOB than exposure to female Wistar urine. As for the mitral/tufted cell layer, the density of Fos-ir cells in the rostral portion (Gi2alpha positive) of all regions of the AOB was about twice as high as that in the caudal portion when male urine was given. The distribution pattern of Fos-ir cells in response to female urine was not identical to that in response to male urine. That is, the density of Fos-ir cells in the caudal portion was slightly larger than that in the rostral portion in the lateral region, while in other regions the density in the rostral portion was higher than that in the caudal portion. It is likely that information from different pheromones is transmitted to the higher brain regions through the different regions of the AOB. PMID- 10383615 TI - Horizontal cells of the rabbit retina are non-selectively connected to the cones. AB - Mammalian horizontal cells have generally been assumed to be spectrally non selective in their cone contacts until recently, when specific contacts have been found for some species. The rabbit retina is frequently studied as a representative of dichromatic mammalian retinae. These are the reasons for elucidating the connections of the two types of horizontal cells (A-HCs and B HCs) with the green-sensitive and blue-sensitive cones of the rabbit retina. Individual A-HCs and B-HCs were revealed by Lucifer Yellow injections, the total cone population overlying them was stained using peanut agglutinin, and the blue cones among these were identified by the antiserum JH 455 against blue cone opsin. Both A-HCs and B-HCs indiscriminately contact the two cone types available. This holds for the green cone-dominated dorsal retina and the blue cone-dominated ventral retina. No evidence was found for a third, potentially blue cone-selective, horizontal cell type [postulated by Famiglietti, E. V. (1990) Brain Res., 535, 174-179]. PMID- 10383616 TI - Stretch-activated cation channels of leech neurons: characterization and role in neurite outgrowth. AB - The goal of this study was to characterize the stretch-activated ion channels (SACs) of adult identified neurons of the leech Hirudo medicinalis and to test the role of SACs in neurite outgrowth of isolated cells. Using cell-attached patch recording, we established that SACs are densely distributed in the growth cone membrane of cultured neurons. In excised patches, we found that these channels are permeable to Ca2+, as well as to monovalent cations. The channels are blocked by the extracellular application of gadolinium (Gd3+), amiloride and gentamicin. Amiloride and gentamicin, respectively, induce a partial and complete voltage-dependent block. Time-lapse video recordings of neurite outgrowth from single cultured neurons were used to study the effects of blocking SACs with gentamicin. Within 20 h of plating in the presence of the aminoglycoside, the total length of neuronal arborization was significantly greater than that measured in its absence. The amount of assembled axon per unitary surface area remained constant over 40 h and did not differ significantly with or without gentamicin. Our findings show that SACs of leech neurons admit Ca2+, are densely distributed in the growth cone membrane and exhibit typical pharmacological features of mechanotransducer ion channels. In addition, our data suggest that these cation channels participate in the early interaction between growing neurites and culture substrate. PMID- 10383617 TI - Hu-Bcl-2 transgenic mice with supernumerary neurons exhibit timing impairment in a complex motor task. AB - Programmed neuronal cell death is common during development, and is thought to be important in the elimination of errors in axonal projection, cell position and sculpting of neuronal circuits. However, the potential importance of programmed cell death for complex behaviour in the adult animal has never been addressed. We studied motor abilities in a strain of transgenic mice with neuronal overexpression of the human Bcl-2 protein, which have supernumerary neurons due to reduced developmental cell death. Our results show that these mice have a clear deficiency in fine timing of motor coordination without impairment of basic motor functions. This is the first indication that altered developmental cell death and the consequent neuronal surplus can impair complex behaviour in the adult animal. PMID- 10383618 TI - Acute application of NGF increases the firing rate of aged rat basal forebrain neurons. AB - Nerve growth factor (NGF) has been widely used in animal models to ameliorate age related neurodegeneration, but it cannot cross the blood-brain barrier (BBB). NGF conjugated to an antibody against the transferrin receptor (OX-26) crosses the BBB and affects the biochemistry and morphology of NGF-deprived basal forebrain neurons. The rapid actions of NGF, including electrophysiological effects on these neurons, are not well understood. In the present study, two model systems in which basal forebrain neurons either respond dysfunctionally to NGF (aged rats) or do not have access to target-derived NGF (intraocular transplants of forebrain neurons) were tested. One group of transplanted and one group of aged animals received unconjugated OX-26 and NGF comixture as a control, while other groups received replacement NGF in the form of OX-26-NGF conjugate during the 3 months preceding the electrophysiological recording session. Neurons from animals in both the transplanted and aged control groups showed a significant increase in firing rate in response to acute NGF application, while none of the conjugate treated groups or young intact rats showed any response. After the recordings, forebrain transplants and aged brains were immunocytochemically stained for the low-affinity NGF receptor. All conjugate treatment groups showed significantly greater staining intensity compared to controls. These data from both transplants and aged rats in situ indicate that NGF-deprived basal forebrain neurons respond to acute NGF with an increased firing rate. This novel finding may have importance even for long-term biological effects of this trophic factor in the basal forebrain. PMID- 10383619 TI - Regulation of the release of 5-hydroxytryptamine in the median raphe nucleus of the rat by catecholaminergic afferents. AB - The present study was conducted in order to examine the influence of catecholaminergic afferents on the release of serotonin in the median raphe nucleus in vivo. To this aim, selective dopamine D1 and D2, and alpha1- and alpha2-adrenergic agonists and antagonists were administered locally (1, 10 and 100 microM) through a dialysis probe implanted in the median raphe nucleus of freely moving rats. The D1 and D2 agonists, (+/-)-1-phenyl-2,3,4, 5-tetrahydro (1H)-3-benzazepine-7,8-diol (SKF-38393) and quinpirole, respectively, and the D1 and D2 antagonists, R-(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4, 5 tetrahydro-1H-3-benzazepine (SCH-23390) and raclopride, respectively, did not alter the release of serotonin in the median raphe nucleus. The alpha1 adrenoceptor agonist phenylephrine did not modify the release of serotonin in this nucleus, although an increased release was observed when the more potent alpha1-adrenoceptor agonist cirazoline was used. In contrast, the alpha1 adrenoceptor antagonist prazosin reduced the release of 5-hydroxytryptamine (5 HT) in a concentration-dependent manner. The release of 5-HT was also reduced by the alpha2-adrenoceptor agonist clonidine and increased by the alpha2 adrenoceptor antagonist 2-methoxy-idazoxan (RX821002). These results indicate that the release of serotonin in the median raphe nucleus does not appear to be regulated by dopaminergic afferents through the activation of dopamine D1 or D2 receptors. On the contrary, it is suggested that endogenous noradrenaline exerts a direct tonic stimulatory control on the release of serotonin through alpha1 adrenoceptors, and an indirect tonic inhibitory influence through alpha2 adrenoceptors located probably in noradrenergic nerve terminals within the median raphe nucleus. PMID- 10383620 TI - Neuroendocrine responses to an emotional stressor: evidence for involvement of the medial but not the central amygdala. AB - The amygdala plays a pivotal role in the generation of appropriate responses to emotional stimuli. In the case of emotional stressors, these responses include activation of the hypothalamic-pituitary-adrenal (HPA) axis. This effect is generally held to depend upon the central nucleus of the amygdala, but recent evidence suggests a role for the medial nucleus. In the present study, c-fos expression, amygdala lesion and retrograde tracing experiments were performed on adult rats in order to re-evaluate the role of the central as opposed to the medial amygdala in generating neuroendocrine responses to an emotional stressor. Brief restraint (15 min) was used as a representative emotional stressor and was found to elicit c-fos expression much more strongly in the medial than central nucleus of the amygdala; relatively few Fos-positive cells were seen in other amygdala nuclei. Subsequent experiments showed that ibotenic acid lesions of the medial amygdala, but not the central amygdala, greatly reduced restraint-induced activation of cells of the medial paraventricular nucleus, the site of the tuberoinfundibular corticotropin-releasing factor cells that constitute the apex of the HPA axis. Medial amygdala lesions also reduced the activation of supraoptic and paraventricular nucleus oxytocinergic neurosecretory cells that commonly accompanies stress-induced HPA axis activation in rodents. To assess whether the role of the medial amygdala in the control of neuroendocrine cell responses to emotional stress might involve a direct projection to such cells, retrograde tracing of amygdala projections to the paraventricular nucleus was performed in combination with Fos immunolabelling. This showed that although some medial amygdala cells activated by exposure to an emotional stressor project directly to the paraventricular nucleus, the number is very small. These findings provide the first direct evidence that it is the medial rather than the central amygdala that is critical to hypothalamic neuroendocrine cell responses during an emotional response, and also provide the first evidence that the amygdala governs oxytocin as well as HPA axis responses to an emotional stressor. PMID- 10383621 TI - Optic flow illusion and single neuron behaviour reconciled by a population model. AB - Radial patterns of optic flow contain a centre of expansion that indicates the observer's direction of self-movement. When the radial pattern is viewed with transparently overlapping unidirectional motion, the centre of expansion appears to shift in the direction of the unidirectional motion [Duffy, C.J. & Wurtz, R.H. (1993) Vision Res., 33, 1481-1490]. Neurons in the medial superior temporal (MST) area of monkey cerebral cortex are thought to mediate optic flow analysis, but they do not shift their responses to parallel the illusion created by transparent overlap. The population-based model of optic flow analysis proposed by Lappe and Rauschecker replicates the illusory shift observed in perceptual studies [Lappe, M. & Rauschecker, J.P. (1995) Vision Res., 35, 1619-1631]. We analysed the behaviour of constituent neurons in the model, to gain insight into neuronal mechanisms underlying the illusion. Single model neurons did not show the illusory shift but rather graded variations of their response specificity. The shift required the aggregate response of the population. We compared the model's predictions about the behaviour of single neurons with the responses recorded from area MST. The predicted distribution of overlap effects agreed with that observed in area MST. The success of the population-based model in predicting the illusion and the neuronal behaviour suggests that area MST uses the graded responses of single neurons to create a population response that supports optic flow perception. PMID- 10383622 TI - Postnatal development of GABAB receptor-mediated modulation of voltage-activated Ca2+ currents in mouse brain-stem neurons. AB - GABAB receptors modulate respiratory rhythm generation in adult mammals. However, little is currently known of their functional significance during postnatal development. In the present investigation, the effects of GABAB receptor activation on voltage-activated Ca2+ currents were examined in rhythmically active neurons of the pre-Botzinger complex (PBC). Both low- (LVA) and high voltage-activated (HVA) Ca2+ currents were present from the first postnatal day (P1). The density of LVA Ca2+ currents increased during the first week, whilst the density of HVA Ca2+ currents increased after the first week. In the second postnatal week, the HVA Ca2+ currents were composed of L- (47 +/- 10%) and N-type (21 +/- 8%) currents plus a 'residual' current, whilst there were no N-type currents detectable in the first few days. The GABAB receptor agonist baclofen (30 microM) increased LVA Ca2+ currents (30 +/- 11%) at P1-P3, but it decreased the currents (35 +/- 11%) at P7-P15 without changing its time course. At all ages, baclofen (30 microM) decreased the HVA Ca2+ currents by approximately 54%. Threshold of baclofen effects on both LVA and HVA Ca2+ currents was 5 microM at P1-P3 and lower than 1 microM at P7-P15. The effect of baclofen was abolished in the presence of the GABAB receptor antagonist CGP 55845A (50 nM). We conclude that both LVA and HVA Ca2+ currents increased postnatally. The GABAB receptor mediated modulation of these currents undergo marked developmental changes during the first two postnatal weeks, which may contribute essentially to modulation of respiratory rhythm generation. PMID- 10383623 TI - Release of endogenous dopamine in cultured mesencephalic neurons: influence of dopaminergic agonists and glucocorticoid antagonists. AB - Several electrochemical techniques allow the measurement of dopamine release in freely moving animals and brain slices. In this report, we applied one of these techniques, coulometry, coupled to high-performance liquid chromatography (HPLC), to the study of dopamine release in primary cultures of embryonic mesencephalic dopaminergic neurons. Between day 9 and 33 of culture, concentrations of dopamine, above the detection threshold, were found in the incubation buffer (Krebs ringer buffer, KRB). Concentrations of dopamine in the incubation buffer reflected neuronal release as they were: (i) positively correlated with the number of tyrosine hydroxylase-positive dopamine neurons in the culture; (ii) tetrodotoxin (TTX) sensitive and Ca2+ dependent; (iii) increased by a depolarizing stimulus, e.g. K+ (20 mM), or by the indirect dopamine agonists amphetamine and cocaine; (iv) decreased by a hyperpolarizing stimulus, e.g. the dopamine D2-like receptor agonist quinpirole. Dopamine release in this model was also sensitive to the manipulation of glucocorticoids, potent modulators of dopamine release in vivo. Long-term treatment of the cell cultures with RU 39305, a selective antagonist of glucocorticoid receptors (GR), but not with spironolactone, a selective antagonist of mineralocorticoid receptors (MR), dose dependently decreased K+-stimulated dopamine release. In conclusion, these results demonstrate an in vitro model that allows the studying of the release of endogenous dopamine in cell cultures and the effects of glucocorticoid hormones on the release dynamics. PMID- 10383624 TI - Native human neocortex release-regulating dopamine D2 type autoreceptors are dopamine D2 subtype. AB - Dopamine (DA) autoreceptors expressed at DA nerve terminals regulate DA release. Considerable evidence has indicated that, in rodents, these autoreceptors belong to the D2 type of the DA receptor family, which, in turn, comprises the D2, D3 and D4 subtypes. We investigated here, for the first time, the subclassification of native human DA autoreceptors by studying the release of [3H]DA evoked by electrical stimulation in fresh human neocortical slices. The results have been compared with those obtained in three animal systems: rat neocortical and striatal slices and rat mesencephalic neuronal cultures. In human neocortical slices, the D2/D3 receptor agonist quinpirole (1 nM-10 microM) inhibited tritium release with a calculated EC50 of 17 nM and a maximal inhibition of approximately 75% reached at 1 microM. In the presence of the D2/D3 receptor antagonist (-) sulpiride (0.1 and 1 microM), the concentration-response curve of quinpirole was shifted to the right, and the apparent pA2 mean value was 8.5 (8.14-8.77); on the other hand, the inhibitory effects of quinpirole were not affected by the D3 receptor-selective antagonist [7-N,N-dipropylamino-5,6,7, 8-tetrahydro naphtho(2,3b) dihydro,2,3-furane] (S 14297) and the D4 receptor-selective antagonist 3-(4-[4-chlorophenyl]piperazin-1-yl)-methyl-1H-pyrrolo [2,3-b]pyridine (L-745,870) (0.01-1 microM in each case). Superimposable results have been obtained when the release was elicited from rat striatal slices or dopamine mesencephalic neurons in culture, whereas quantitative differences emerged in the case of rat cortical slices. It is concluded that in human brain, as well as in rat brain, the release of DA in the terminal region of midbrain dopaminergic neurons is regulated through autoreceptors of the D2 subtype. PMID- 10383625 TI - Reduced hippocampal LTP in mice lacking a presynaptic protein: complexin II. AB - The SNAP receptor (SNARE) complex is a core complex specialized for synaptic vesicle exocytosis, and the binding of SNAPs to the complex is an essential step for neurotransmitter release. Complexin I and II have been identified as SNARE complex-associated proteins. Importantly, complexins compete with alpha-SNAP for binding to the complex, suggesting that complexins may modulate neurotransmitter release process. To examine this possibility and to understand the physiological function of complexins, we generated complexin II knockout mice. The complexin-II deficient mice (-/-) were viable and fertile, and appeared normal. Electrophysiological recordings in the mutant hippocampus showed that ordinary synaptic transmission and paired-pulse facilitation, a form of short-term synaptic plasticity, were normal. However, long-term potentiation (LTP) in both CA1 and CA3 regions was impaired, suggesting that complexin II may not be essential for synaptic vesicle exocytosis, but it does have a role in the establishment of hippocampal LTP. PMID- 10383626 TI - Inhibition of glial scarring in the injured rat brain by a recombinant human monoclonal antibody to transforming growth factor-beta2. AB - The transforming growth factor-betas (TGF-betas) are potent fibrogenic factors implicated in numerous central nervous system (CNS) pathologies in which fibrosis and neural dysfunction are causally associated. In this study, we aim to limit the fibrogenic process in a model of CNS scarring using a recombinant human monoclonal antibody, derived from phage display libraries and specific to the active form of the TGF-beta2 isoform. The implicit inference of the work was that, as such antibodies are potential pharmacological agents for the treatment of human CNS fibrotic diseases, validation of efficacy in a mammalian animal model is a first step towards this end. Treatment of cerebral wounds with the anti-TGF-beta2 antibody led to a marked attenuation of all aspects of CNS scarring, including matrix deposition, formation of an accessory glial-limiting membrane, inflammation and angiogenesis. For example, in the wound, levels of: (i) the connective tissue components fibronectin, laminin and chondroitin sulphate proteoglycan; and (ii) wound-responsive cells including astrocytes and macrophages/microglia, were markedly reduced. Our findings suggest that such synthetic anti-fibrotic TGF-beta antibodies are potentially applicable to a number of human CNS fibrotic diseases to arrest the deposition of excessive extracellular matrix components, and maintain and/or restore functional integrity. PMID- 10383627 TI - Calpain activation and inhibition in organotypic rat hippocampal slice cultures deprived of oxygen and glucose. AB - It has been suggested that, after ischaemia, activation of proteases such as calpains could be involved in cytoskeletal degradation leading to neuronal cell death. In vivo, calpain inhibitors at high doses have been shown to reduce ischaemic damage and traumatic brain injury, however, the relationship between calpain activation and cell death remains unclear. We have investigated the role of calpain activation in a model of ischaemia based on organotypic hippocampal slice cultures using the appearance of spectrin breakdown products (BDPs) as a measure of calpain I activation. Calpain I activity was detected on Western blot immediately after a 1-h exposure to ischaemia. Up to 4 h post ischaemia, BDPs were found mainly in the CA1 region and appeared before uptake of the vital dye propidium iodide (PI). 24 h after the insult, BDPs were detected extensively in CA1 and CA3 pyramidal cells, all of which was PI-positive. However, there were many more PI-positive cells that did not have BDPs, indicating that the appearance of BDPs does not necessarily accompany ischaemic cell death. Inhibition of BDP formation by the broad-spectrum protease inhibitor leupeptin was not accompanied by any neuroprotective effects. The more specific and more cell-permeant calpain inhibitor MDL 28170 had a clear neuroprotective effect when added after the ischaemic insult. In contrast, when MDL 28170 was present throughout the entire pre- and post-incubation phases, PI labelling actually increased, indicating a toxic effect. These results suggest that calpain activation is not always associated with cell death and that, while inhibition of calpains can be neuroprotective under some conditions, it may not always lead to beneficial outcomes in ischaemia. PMID- 10383628 TI - Modulation of steroidal levels by adrenalectomy/castration and inhibition of neurosteroid synthesis enzymes affect sigma1 receptor-mediated behaviour in mice. AB - The interaction between neurosteroids and sigma1 (sigma1) receptors may be of therapeutic interest during physiological or pathological ageing, particularly concerning their neuromodulatory role on cognitive functions. Neurosteroids modulate memory processes through a mechanism involving interactions with GABAA, N-methyl-D-aspartate and/or sigma1 receptors. To measure the contribution of endogenous neurosteroid levels to the antiamnesic effects of sigma1 agonists, we investigated the effects of inhibitors of key enzymes involved in neurosteroid synthesis, in adrenalectomized/castrated (AdX/CX) mice to avoid the effect of circulating steroids. Trilostane, a 3beta-hydroxysteroid-deshydrogenase inhibitor, blocks the pregnenolone to progesterone conversion and leads to a decrease of progesterone. Finasteride, a 5alpha-reductase inhibitor, blocks the progesterone to 5alpha-pregnane-3,20-dione conversion and leads to an accumulation of progesterone. The in vivo binding of (+)-[3H]SKF-10 047 to sigma1 sites was measured in the mouse hippocampus and cortex. The attenuating effect of the selective sigma1 agonist PRE-084 (0.1-3 mg/kg) against dizocilpine (0.15 mg/kg)-induced learning impairment was examined using spontaneous alternation behaviour, step-down passive avoidance and place learning in the elevated plus maze. The in vivo (+)-[3H]SKF-10 047 binding appeared significantly increased in AdX/CX mice and after trilostane treatment (10 mg/kg twice a day, 7 days), compared with sham-operated animals. The finasteride treatment (25 mg/kg, 7 days) significantly decreased binding levels. The learning deficits induced by dizocilpine were not affected by the treatments. The antiamnesic effect of PRE 084 was facilitated in AdX/CX mice and even more after trilostane treatment, as several parameters for animals treated with both PRE-084 and dizocilpine returned to control values. The PRE-084 effect was blocked after finasteride. These results confirmed that endogenous neurosteroidal levels modulate sigma1 receptor mediated behaviour directly, and revealed that, among neurosteroids, progesterone may be the main modulator of sigma1 receptors. PMID- 10383629 TI - Li+ and muscarine cooperatively enhance the cationic tail current in rat cortical pyramidal cells. AB - Li+ is known to facilitate the onset of status epilepticus induced by cholinergic stimulation, although the underlying mechanisms are not clear. Under whole-cell current clamp conditions with a CsCl-based internal solution, cortical pyramidal cells display a single plateau-spike followed by a slow depolarizing afterpotential (DAP) in response to injection of a short current pulse. However, the same current pulse generated a burst of plateau-spikes after application of Li+ (2 mM) and muscarine (10 microM). As similar bursts of plateau-spikes were generated through an enhancement of the slow DAP when [K+]o was raised (Kang et al. 1998), we have investigated the effects of Li+ and muscarine on the Ca2+ dependent cationic current underlying the slow DAP, measured as the slow tail current evoked after the offset of depolarizing voltage pulses. Muscarine enhanced the amplitudes of both early and late components of the slow tail current. This effect of muscarine was markedly potentiated by Li+, while Li+ by itself affected the slow tail current only slightly. Intracellular application of heparin (0.5-1 mg/mL) suppressed the effect of muscarine in the presence of Li+. These results suggest that inositol-trisphosphate-induced Ca2+ release is involved in the cooperative enhancement of the slow tail current, and this cooperation may be one of the mechanisms underlying facilitation of the onset of epilepsy induced by these agents. PMID- 10383630 TI - Expression in mammalian cells and electrophysiological characterization of two mutant Kv1.1 channels causing episodic ataxia type 1 (EA-1). AB - Episodic ataxia type 1 (EA-1) is a rare neurological disorder and was the first ionic channel disease to be associated with defects in a potassium channel. Until now 10 different point mutations in the KCNA1-gene have been reported to cause this disorder. We have investigated the functional consequences of two mutations leading to amino acid substitutions in the first and sixth transmembrane segments of a Kv1.1 channel subunit, by means of the patch-clamp technique; we injected cRNA coding for, respectively, F184C and V408A mutant Kv1.1 channels into mammalian cells and compared the resulting currents with those in the wild-type. The expression levels of F184C and V408A mutant channels relative to that of the wild-type was 38 and 68%, respectively. Since the single-channel conductance of the F184C mutant was similar to that of the wild-type (12 pS) without an apparent change in the maximum open probability, we conclude that the lower expression level in the F184C mutant channels is due to a reduced number of functional channels on the cell surface. F184C activated slower, and at more depolarized potentials, and deactivated faster compared with the wild-type. V408A channels deactivated and inactivated faster compared with the wild-type. Studies with different extracellular cations and tetraethylammonium gave no indication that the pore structure was changed in the mutant channels. Acetazolamide, that is helpful in some patients suffering from EA-1, was without effect on Kv1.1 wild type or mutant channels. This study confirms and extends earlier studies on the functional consequences of Kv1.1 mutations associated with EA-1, in an attempt to understand the pathophysiology of the disease. PMID- 10383631 TI - Neuroprotection of cultured foetal rat hippocampal cells against glucose deprivation: are GABAergic neurons less vulnerable or more sensitive to TCP protection? AB - In the rat brain, hippocampal neurons are particularly sensitive to secondary excitotoxic injury induced by ischaemia or hypoglycaemia. To determine some distinctive features of vulnerability among neuronal phenotypes in the hippocampus following a metabolic insult, we used an in vitro model of mild glucose deprivation. Primary cultures from the rat hippocampus (21 days in vitro) were deprived of glucose for 4 h and then were returned to the standard medium for 24 or 48 h. Survival of the GABAergic neuronal population was evaluated both by measuring [3H]GABA uptake and by counting GAD65-immunostained cells. This was compared with the survival of the total neuronal population evaluated by counting the neurofilament-200-immunostained cells. Glucose deprivation for 4 h followed by a recovery period of 48 h induced a decrease of 59% and 40% in the number of GAD65- and neurofilament-200-immunostained cells, respectively. Thus, GABAergic neurons were slightly more vulnerable to glucose deprivation than the other neurons in the hippocampal cell cultures. When the excitotoxic component of cellular death was blocked in the presence of TCP, an NMDA-antagonist, the survival of GABAergic neurons was almost complete after 48 h of recovery. In contrast, measurements of the release of lactate dehydrogenase in the medium indicated that TCP largely protected hippocampal cells after 24 h but was ineffective after 48 h. This observation was confirmed by immunostaining data which showed that after 48 h TCP did not significantly increase the survival of neurofilament-200-immunostained cells. These results indicate that after glucose deprivation and a recovery period of 48 h, GABAergic neurons in hippocampal cell cultures are not more resistant than other neurons but they are more sensitive to TCP protection. PMID- 10383632 TI - Calcium-dependent inactivation of the monosynaptic NMDA EPSCs in rat hippocampal neurons in culture. AB - The effects of increased dendritic calcium concentration ([Ca2+]i) induced by single action potentials on monosynaptic glutamatergic excitatory postsynaptic currents (EPSCs) were studied in cultured rat hippocampal neurons. To investigate the respective roles of pre- and postsynaptic elements in the depolarization induced NMDAR inactivation, we have performed simultaneous paired whole-cell recordings from monosynaptically connected pre- and postsynaptic hippocampal neurons. We report that the single firing of the postsynaptic neuron did not result in inactivation of the NMDAR-EPSC, whereas a burst of depolarizing steps transiently depressed the NMDAR-EPSCs in both pyramidal cells and interneurons. This effect was mediated by postsynaptic voltage-gated Ca2+ influx, as it was prevented by: (i) buffering postsynaptic [Ca2+]i with 30 mM BAPTA; (ii) removing extracellular Ca2+; or (iii) applying Cd2+o (100 microM), a voltage-gated calcium channel blocker. It does not involve presynaptic mechanisms as it selectively affected NMDA but not alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptor-mediated EPSCs. These results suggest that inactivation of NMDAR channels by voltage-gated Ca influx is a general property of hippocampal neurons, which may play an important role in reducing postsynaptic NMDAR Ca2+ influx that leads to plasticity or excitotoxicity during sustained neuronal activity. PMID- 10383633 TI - Behavioural conditions affecting saccadic eye movements elicited electrically from the frontal lobes of primates. AB - We assessed the effects of varying the time at which electrical stimulation was delivered to the dorsomedial frontal cortex (DMFC) and the frontal eye fields (FEF) relative to the onset of a visual target. Monkeys were required to fixate the visual target to obtain a drop of apple juice as reward. We found that the probability of eliciting saccades increased with increases in the delay of electrical stimulation relative to target onset. Also, the current threshold to evoke saccades decreased as electrical stimulation was delivered later following target onset. There were major differences in the magnitude of this effect with stimulation of the DMFC versus the FEF. The current threshold to evoke saccades from the DMFC was 16 times greater when electrical stimulation was delivered 200 ms after target onset as compared to when it was delayed 200 ms after target offset. In contrast, the current threshold to evoke saccades from the FEFs was only three times greater when stimulation was delivered under similar conditions. These results suggest that the FEF are more closely connected with the saccade generator for the execution of saccadic eye movements than is the DMFC, even though both regions have direct projections to brainstem oculomotor centres. PMID- 10383634 TI - Differential regulation of distinct phenotypic features of serotonergic neurons by bone morphogenetic proteins. AB - Bone morphogenetic proteins (BMPs), growth and differentiation factor 5 (GDF5) and glial cell line-derived neurotrophic factor (GDNF) are members of the transforming growth factor-beta superfamily that have been implicated in tissue growth and differentiation. Several BMPs are expressed in embryonic and adult brain. We show now that BMP-2, -6 and -7 and GDF5 are expressed in the embryonic rat hindbrain raphe. To start to define roles for BMPs in the regulation of serotonergic (5-HT) neuron development, we have generated serum-free cultures of 5-HT neurons isolated from the embryonic (E14) rat raphe. Addition of saturating concentrations (10 ng/mL) of BMP-6 and GDF5 augmented numbers of tryptophan hydroxylase (TpOH) -immunoreactive neurons and cells specifically taking up 5, 7 dihydroxytryptamine (5,7-DHT) by about two-fold. Alterations in 5-HT neuron numbers were due to the induction of serotonergic markers rather than increased survival, as shown by the efficacy of short-term treatments. Importantly, BMP-7 selectively induced 5, 7-DHT uptake without affecting TpOH immunoreactivity. BMP 6 and -7 also promoted DNA synthesis and increased numbers of cells immunoreactive for vimentin and glial fibrillary acidic protein (GFAP). Pharmacological suppression of cell proliferation or glial development abolished the induction of serotonergic markers by BMP-6 and -7, suggesting that BMPs act indirectly by stimulating synthesis or release of glial-derived serotonergic differentiation factors. Receptor bodies for the neurotrophin receptor trkB, but not trkC, abolished the BMP-mediated effects on serotonergic development, suggesting that the glia-derived factor is probably brain-derived neurotrophic factor (BDNF) or neurotrophin-4. In support of this notion, we detected increased levels of BDNF mRNA in BMP-treated cultures. Together, these data suggest both distinct and overlapping roles of several BMPs in regulating 5-HT neuron development. PMID- 10383635 TI - Long-term effects of methylprednisolone following transection of adult rat spinal cord. AB - Clinically, high-dose treatment with the glucocorticosteroid, methylprednisolone (MP), within 8 h after spinal cord injury, has been shown to improve neurological recovery. The current standard of care is to administer MP as a bolus of 30 mg/kg followed by a 23-h infusion of 5.4 mg/kg/h to spinal cord injured patients. To better understand the role of MP in neuroprotection, we have studied how MP administration affects macrophage accumulation, tissue loss, and axonal dieback at 1, 2, 4 and 8 weeks after a complete transection of the eighth thoracic spinal cord in the adult rat. A 30 mg/kg dose of MP was administered intravenously at 5 min, and 2 and 4 h after injury. The number of ED1 (antibody against microglia/macrophages) -positive cells was quantified in a 500-micrometer-wide strip of tissue directly adjacent and parallel to the transection. At all time points, MP treatment led to a significant decrease in the number of ED1-positive cells in both rostral and caudal stumps. Over the 2-month post-transection period, the average MP-induced reduction in the number of ED1-positive cells was 82% in the rostral cord stump and 66% in the caudal stump. Using a computerized image analysis system, it was observed that MP treatment resulted in a significant reduction in tissue loss in both cord stumps at 2, 4 and 8 week post injury. Over the 2-month post-lesion period, the average MP-induced reduction in tissue loss in the caudal cord stump was higher than that in the rostral stump; 48 versus 37%, respectively. Immunostaining for neurofilaments and growth associated protein-43 (GAP-43) revealed the presence of numerous axons near and in the lesion site. Anterograde neuronal tracing with biotinylated dextran amine showed that, in MP-treated animals, dieback of vestibulospinal fibres, but not of corticospinal fibres, was significantly diminished at all time points studied. In addition, with MP administration, 1 and 2 weeks after injury, an increase in the number of vestibulospinal fibres was found at 1 and 2 mm from the transection, suggesting transient regenerative sprouting of these fibres. The results demonstrate that treatment with MP shortly after spinal cord transection in the adult rat led to a long-term reduction of ED1-positive cells and spinal tissue loss, reduced dieback of vestibulospinal fibres, and a transient sprouting of vestibulospinal fibres near the lesion at 1 and 2 weeks post-lesion. The possible relationships between the inflammatory changes, spinal tissue sparing, and axonal survival and sprouting are complex and need to be further explored. PMID- 10383636 TI - Rapid glucocorticoid effects on excitatory amino acid levels in the hippocampus: a microdialysis study in freely moving rats. AB - Glucocorticoids can rapidly affect neuronal function and behaviour in mammals. Several studies have suggested the possible existence of rapid, non-genomic effects of glucocorticoids in the hippocampus. To investigate whether glucocorticoids could affect neurotransmission in the hippocampus through rapid, non-genomic mechanisms, we studied the effects of acute glucocorticoid administration on extracellular amino acid levels in the CA1 area of the hippocampus. By means of microdialysis on freely moving rats, we observed that an intraperitoneal injection of corticosterone (2.5 mg/kg) induced a rapid (within 15 min) and transient (returning to basal levels by 35-45 min) increase in extracellular aspartate and glutamate levels ( approximately 155-160%), both in sham-operated and adrenalectomized rats. These effects occurred in parallel with a rise in corticosterone concentration, also detected by microdialysis, in this hippocampal area. Intrahippocampal perfusion of corticosterone by retrodialysis also produced the same fast and reversible effects on excitatory amino acid (EAA) levels. Extracellular concentrations of taurine and gamma-aminobutyric acid (GABA) were unchanged after intrahippocampal glucocorticoid administration. This corticosterone-mediated rise in EAA levels was not inhibited by the presence of specific antagonists for the two types of intracellular corticosteroid receptors, nor by a protein synthesis inhibitor, anisomycin. Perfusion of dexamethasone, a synthetic glucocorticoid, elicited a similar effect to that observed with corticosterone treatment in all studied cases. However, non-glucocorticoid steroids did not affect amino acid transmission in this hippocampal area. These results indicate that glucocorticoids induce a rapid and transient increase in hippocampal EAA levels in vivo that might be exerted through a novel non-genomic mechanism of action. PMID- 10383637 TI - Tenascin-R interferes with integrin-dependent oligodendrocyte precursor cell adhesion by a ganglioside-mediated signalling mechanism. AB - Oligodendrocyte (OL) lineage progression is characterized by the transient expression of the disialoganglioside GD3 by OL precursor (preOL) cells followed by the sequential expression of myelin-specific lipids and proteins. Whereas GD3+ preOLs are highly motile cells, the migratory capacity of OLs committed to terminal differentiation is strongly reduced, and we have recently shown that the extracellular matrix protein tenascin-R (TN-R) promotes the stable adhesion and differentiation of O4+ OLs by a sulphatide-mediated autocrine mechanism (O4 is a monoclonal antibody recognizing sulphatides/seminolipids expressed by OLs and in myelin). Using culture conditions that allow the isolation of mouse OLs at distinct lineage stages, here we demonstrate that TN-R is antiadhesive for GD3+ preOLs and inhibits their integrin-dependent adhesion to fibronectin (FN) by a disialoganglioside-mediated signalling mechanism affecting the tyrosine phosphorylation of the focal adhesion kinase. This responsive mechanism appears to be common to various cell types expressing disialogangliosides as: (i) disialogangliosides interfered with the inhibition of cell adhesion of different neural and non-neural cells on substrata containing TN-R and FN or RGD-containing FN fragments. TN-R interacted specifically with disialoganglioside-expressing cells or immobilized gangliosides, and ganglioside treatment of TN-R substrata resulted in a delayed preOL cell detachment as a function of time. We conclude that OL response to one and the same signal in the extracellular matrix critically depends on the molecular repertoire expressed by OLs at different lineage stages and could thus define their final positioning. PMID- 10383638 TI - Reversal of glutamate excitotoxicity by activation of PKC-associated metabotropic glutamate receptors in cerebellar granule cells relies on NR2C subunit expression. AB - Stimulation of metabotropic glutamate receptors (mGluRs) belonging to group I has been found to reduce N-methyl-D-aspartate (NMDA) receptor function in terms of both intracellular calcium concentration ([Ca2+]i) rise and neurotoxicity in cultured cerebellar granule cells. In the present study, we investigated whether the mGluR-elicited modulation of glutamate responses might rely on the heteromeric composition of NMDA receptor channel. NMDA receptors consist of two distinct groups of subunits: NR1, that is ubiquitously in the receptor complexes; and NR2A-D, that differentiate and potentiate NMDA receptor responses by assembling with NR1. Among NR2 subunits, only NR2A and NR2C mRNAs and relative proteins are detected in cerebellar granule cells at 10 days in vitro. To dissect the involvement of the two different subunits in making the NMDA receptor channel sensitive to modulation by group I mGluR agonists, expression of the NR2C subunit was prevented by treating the cells with specific antisense oligodeoxynucleotide (ODN). The capability of the mGluR agonists, trans-1-amino-cyclopentane-1,3 dicarboxylic acid (tACPD, 100 microM) or 3 hydroxyphenylglycine (3HPG, 100 microM), and the protein kinase C (PKC) activator, 4beta-phorbol-12,13-dibutyrate (PDBu, 1 microM), to inhibit the function of resultant NMDA receptors was then evaluated. We found that depletion of the NR2C subunit abolished the inhibitory effect of group I mGluR stimulation on glutamate-induced [Ca2+]i rise and neurotoxicity. The antisense ODN treatment also prevented the inhibitory effect of PDBu on glutamate responses. Conversely, in NR2C-lacking neurons, both group I mGluRs and PKC stimulation enhanced NMDA receptor-mediated effects. The present findings indicate that the capability of PKC-associated mGluRs to modulate native NMDA receptor function relies on the heteromeric configuration of the receptor channel complex. Particularly, expression of the NR2C subunit is required to make the NMDA receptor sensitive to inhibitory modulation by mGluRs or PKC activation. PMID- 10383639 TI - The effect of etomidate on sensory transmission in the dorsal column pathway in the urethane-anaesthetized rat. AB - The effect of Etomidate, a general anaesthetic, on sensory afferent transmission was measured in the dorsal column pathway in urethane-anaesthetized rats. Extracellular recordings were made of peripherally evoked responses by single cells in the cuneate nucleus, ventroposterolateral nucleus of the thalamus and laminae IV-VI of the primary somatosensory cortex. Cortical mass responses were also recorded. In further experiments, cortical mass responses were evoked antidromically by stimulation in the pyramidal tract. The effect of incremental administration of Etomidate on evoked responses was recorded. These results are compared with the previously reported effects of urethane, a 'conventional' anaesthetic. Etomidate did not alter cuneate or ventroposterolateral thalamic cell responses but it caused a dose-dependent reduction in cortical cell responsiveness. It failed to alter antidromically evoked cortical mass responses. Etomidate differs from the majority of anaesthetics, which act in the thalamus, and appears to cause perturbation at the cortical level. PMID- 10383640 TI - Architecture and connections of retrosplenial area 30 in the rhesus monkey (Macaca mulatta). AB - Because of the sharp curvature of the retrosplenial region around the splenium of the corpus callosum, standard coronal sections are not appropriate for architectonic analysis of its posteroventral part. In the present study, examination of the posteroventral retrosplenial region of the rhesus monkey in sections that were orthogonal to its axis of curvature (and therefore appropriate for architectonic analysis) has permitted definition of its architecture and precise extent. This analysis demonstrated that areas 29 and 30 of the retrosplenial cortex, as well as adjacent area 23 of the posterior cingulate cortex, extend together as an arch around the splenium of the corpus callosum and maintain their topographical relationship with one another throughout their entire course. Injections of anterograde and retrograde tracers confined to retrosplenial area 30 revealed that this area has reciprocal connections with adjacent areas 23, 19 and PGm, with the mid-dorsolateral part of the prefrontal cortex (areas 9, 9/46 and 46), with multimodal area TPO in the superior temporal sulcus, as well as the posterior parahippocampal cortex, the presubiculum and the entorhinal cortex. There are also bidirectional connections with the lateroposterior thalamic nucleus, as well as the laterodorsal and the anteroventral limbic thalamic nuclei. The connectivity of area 30 suggests that it may play a role in working memory processes subserved by the mid-dorsolateral frontal cortex in interaction with the hippocampal system. PMID- 10383642 TI - Immunological evidence that the beta isoform of Ca2+/calmodulin-dependent protein kinase IV is a cerebellar granule cell-specific product of the CaM kinase IV gene. AB - Ca2+/calmodulin-dependent protein kinase IV (CaM kinase IV) exists as two monomeric isoforms, alpha and beta. In this study, we raised an antibody against the beta isoform and provided immunohistochemical evidence for specific expression of the beta isoform in cerebellar granule cells as a single gene derived translational product distinct from the alpha isoform. Immunohistochemical examination showed that the beta-immunoreactivity was confined to the nuclei of the cerebellar granule cells, in contrast to the more widespread immunoreactivity for the alpha isoform in both nuclei and cytoplasm of the cerebellar granule cells and many other neurons with dominant nuclear localization. In developing cerebella, the beta-immunoreactivity gradually appeared in the internal granule cells during the postnatal 2nd and 3rd weeks, while the alpha-immunoreactivity had already appeared in the internal granule cells in the 1st postnatal week. Unlike the alpha isoform, beta-immunoreactivity was not detected in the Purkinje cells at any developmental stages. The differential expression of the alpha and beta isoforms suggests that each isoform may be involved in different cerebellar functions. PMID- 10383641 TI - Allatostatin modulates skeletal muscle performance in crustaceans through pre- and postsynaptic effects. AB - Allatostatins, originally identified in insects as peptide inhibitors of juvenile hormone biosynthesis, are regarded as potent inhibitory regulators of intestinal muscles in insects and crustaceans. However, accumulating data indicate that allatostatins might also be involved in modulation of skeletal neuromuscular events. We show that most ganglia of two isopod crustaceans (Idotea baltica and I. emarginata) contain pairs of large, allatostatin-immunoreactive motor neurons which supply several segmental muscles. Among them are the dorsal extensor muscles, of which some fibres receive immunoreactive, varicose innervation. We demonstrate, on identified muscle fibres, that allatostatin exerts a twofold inhibitory effect: it reduces contractions of single voltage-clamped fibres, and it decreases the amplitude of evoked excitatory junctional currents recorded from individual release boutons. No change in excitation-contraction threshold or in passive membrane parameters was observed. As the amplitude of miniature currents generated by spontaneously released single transmitter quanta was not changed, the inhibitory effect of the peptide on junctional currents must be of presynaptic origin. Supportive results were obtained on leg muscles of the crab Eriphia spinifrons, where allatostatin decreased evoked synaptic currents by reducing the mean number of transmitter quanta released by presynaptic depolarization without affecting the amplitudes of currents generated by single quanta. This effect of allatostatin was similar for two functionally different neurons, the slow and the fast closer excitor. The data show that allatostatin occurs in identified motor neurons of Idotea and exerts complementary pre- and postsynaptic modulatory effects which reduce muscle responses. Thus, allatostatin counteracts the effects of another neuropeptide, proctolin, which is also present in Idotea and causes potentiating effects on the same muscle fibres. PMID- 10383643 TI - Astrocytes regulate the developmental appearance of GABAergic and glutamatergic postsynaptic currents in cultured embryonic rat spinal neurons. AB - The effects of astrocytes on the emergence of synaptic transients and excitable membrane properties in cultured, embryonic, rat ventral spinal neurons were studied with electrical and optical recording techniques. Neurons on astrocytes had significantly longer neurites and an accelerated rate of growth in surface membrane during the initial 24 h in culture compared to neurons on poly-D-lysine (PDL). GABAergic (GABA, gamma-aminobutyric acid) and glutamatergic transients appeared spontaneously in co-cultured neurons by 24 h. GABAergic quanta did not appear in neurons on PDL until 4 days in culture, and glutamatergic transients did not emerge until 7 days in culture. Astrocyte-conditioned medium (ACM) partially mimicked the effects of direct astrocytic contact. GABAergic transients appeared by 2 days, and glutamatergic signals by 4 days in neurons on PDL exposed to ACM. All of the spontaneous, synaptic-like transients were eliminated by tetrodotoxin or Ca2+o-free saline, implicating voltage-dependent cation channels in their generation. Astrocytes immediately and significantly increased the density of voltage-dependent Na+ currents compared to neurons on PDL, but by the end of 24 h, Na+ current densities were identical. Electrophysiological and optical recording revealed comparable densities of high-voltage-activated (HVA) Ca2+ currents on both co-cultured neurons and neurons on PDL throughout the first week. However, neurons on astrocytes had significantly greater contributions of P/Q-type currents and lesser contributions of L-type currents beginning at 24 h and continuing for 7 days. The contribution of N-type current was significantly more in co-cultured neurons only at 24 h. Thus, in vitro, astrocytes help to differentiate specific excitable membrane properties in spinal neurons, along with GABAergic and glutamatergic forms of synaptic transmission. PMID- 10383644 TI - Expression of nicotinic acetylcholine receptor subunits in the cerebral cortex in Alzheimer's disease: histotopographical correlation with amyloid plaques and hyperphosphorylated-tau protein. AB - Impairment of cholinergic transmission and decreased numbers of nicotinic binding sites are well-known features accompanying the cognitive dysfunction seen in Alzheimer's disease (AD). In order to elucidate the underlying cause of this cholinoceptive dysfunction, the expression of two pharmacologically different nicotinic acetylcholine receptor (nAChR) subunits (alpha4, alpha7) was studied in the cerebral cortex of Alzheimer patients as compared to controls. Patch-clamp recordings of 14 dissociated neurons of control cortices showed responses suggesting the existence of alpha4- and alpha7-containing functional nAChRs in the human cortex. In cortices of Alzheimer patients and controls, the pattern of distribution and the number of alpha4 and alpha7 mRNA-expressing neurons were similar, whereas at the protein level a decrease in the density of alpha4- and alpha7-expressing neurons of approximately 30% was observed in Alzheimer patients. The histotopographical correlation of nAChR expression with accompanying pathological changes, e.g. accumulation of hyperphosphorylated-tau (HP-tau) protein and beta-amyloid showed that neurons in the vicinity of beta amyloid plaques bore both nAChR transcripts. Neurons heavily labelled for HP-tau, however, expressed little or no alpha4 and alpha7 mRNA. These results point to an impaired synthesis of nAChRs on the protein level as a possible cause of the cholinoceptive deficit in AD. Further investigations need to elucidate whether interactions of HP-tau with nAChR mRNA, or alterations in the quality of alpha4 and alpha7 transcripts give rise to decreased protein expression at the level of individual neurons. PMID- 10383645 TI - Filial imprinting in domestic chicks is associated with spine pruning in the associative area, dorsocaudal neostriatum. AB - Juvenile emotionally modulated learning events are fundamental for the normal development of socio-emotional competence and intellectual capabilities. Filial imprinting in the domestic chick provides a suitable model to investigate the neural mechanisms underlying such juvenile learning events. The forebrain area dorsocaudal neostriatum (Ndc), a multimodal integration area and presumed equivalent to mammalian parietotemporal association cortices, has been shown to be critically involved in this learning process. We investigated whether filial imprinting is associated with changes of synaptic connectivity in the Ndc. Quantitative measurements of spine densities of a large neuron type in the Ndc revealed a massive pruning of spine synapses after filial imprinting. Compared with 7-day-old naive control chicks, imprinted chicks displayed significantly lower spine frequencies on all dendritic segments. Since the average length of the dendritic segments did not change during imprinting, these results can be interpreted as a reduction of the absolute number of spine synapses on this neuron type. In a control region, the primary sensory forebrain area ectostriatum, spine density and dendritic length remained unchanged. These results indicate that synaptic pruning may represent a mechanism of selective synaptic reorganization in higher associative forebrain areas as a fundamental feature of juvenile learning events. PMID- 10383646 TI - Type III intermittency in human partial epilepsy. AB - A rigorous characterization of the dynamic regimes underlying human seizures is needed to understand, and possibly control, the transition to seizure. Intra- or extracranial brain electrical activity was recorded in five patients with partial epilepsy, and the interictal and ictal activity analysed to determine the dynamics of seizures. We constructed first-return one-dimensional maps by fitting the scatter plots of interpeak intervals. The features of the mapping indicated that type III intermittency is the dynamic characteristic of the ictal events. This was confirmed using histograms of the durations of the regular phases during seizures. The intermittent regime explains the abrupt transitions observed during ictal events in terms of transient stabilization of the unstable steady state. PMID- 10383647 TI - Accumulation of murine amyloidbeta42 in a gene-dosage-dependent manner in PS1 'knock-in' mice. AB - The establishment of an animal model with a missense mutation of presenilin-1 (PS1) is an initial step toward understanding the molecular pathogenesis of familial Alzheimer's disease (FAD) and developing therapeutic strategies for the disease. We previously described a Japanese family with FAD caused by the I213T mutation of PS1, in which typical signs and symptoms of Alzheimer's disease were observed at the age of 45 +/- 4.2 years [Hardy, J. (1997) Trends. Neurosci., 20, 154-159; Kamino, K et al. (1996) Neurosci. Lett., 208, 195-198]. Here, we report the establishment of 'knock-in' mice with the I213T PS1 missense mutation. Northern blot and reverse transcription polymerase chain reaction (RT-PCR) analyses showed that the mutated PS1 allele was expressed at the same level as the endogenous PS1 allele, demonstrating that the PS1 missense mutation was successfully introduced into the mouse PS1 locus, and therefore that the situation mimics that in FAD patients bearing PS1 missense mutations. Amyloid beta (Abeta) 42(43) peptide, but not Abeta40 peptide, accumulated in 'knock-in' mice at the age of 16-20 weeks. A clear gene-dosage effect on the increase of Abeta42(43) was observed in 'knock-in' mice: the percentage increase of Abeta42(43) in mice with mutations in both alleles was twice as high as that in mice with a single allele. These results indicate that the level of the mutated PS1 gene expression is likely to be critically involved in the production of highly amyloidogenic Abeta42(43), and confirm that PS1 mutation has an important effect on amyloid precursor protein (APP) processing, in proportion to the level of the expression of the mutant gene. PMID- 10383648 TI - Ectopic expression of the bHLH gene Math1 disturbs neural development. AB - The basic helix-loop-helix gene Math1, a positive regulator of neuronal differentiation, is specifically expressed in the dorsal part of the developing nervous system. To determine the effects of ectopic expression of Math1, we generated two transgenic mouse lines; One carried the Cre recombinase gene under the control of the nestin promoter and enhancer, which direct expression in neural precursor cells, and the other carried the Math1 gene, the expression of which was regulated by the cytomegalovirus (CMV) promoter but interrupted by the stop cassette flanked by loxP sites. In F1 embryos that carried the two transgenes, the stop cassette was removed by Cre recombinase in the developing nervous system, and Math1 expression was ectopically directed from the CMV promoter. We found that these embryos exhibited abnormal morphology of the brain and extensive cell death in the nervous system. These results suggest that ectopic expression of Math1 is toxic to neurons and leads to apoptosis. PMID- 10383649 TI - Requirement for DARPP-32 in mediating effect of dopamine D2 receptor activation. AB - It is well documented that dopamine and dopamine D1 agonists convert the protein phosphatase-1 inhibitor, DARPP-32, from its dephosphorylated, inactive form into its Thr34-phosphorylated, active form, and that these effects on DARPP-32 constitute essential components of the mechanism by which dopamine and D1 agonists achieve their biological effects. In contrast to dopamine and D1 agonists, dopamine D2 agonists dephosphorylate and inactivate DARPP-32. Here we have examined the possibility that the biological effects of dopamine D2 receptor agonists might also involve DARPP-32. For this purpose, we have examined regulation of the activity of the electrogenic ion pump Na+,K+-ATPase, an established target for dopamine signalling. We have found that dopamine D1 agonists and dopamine D2 agonists inhibit Na+,K+-ATPase activity in dissociated cells from the mouse neostriatum and that, in each case, the effect is abolished in cells from mice deficient in DARPP-32. We conclude that DARPP-32 may play an obligatory role in dopaminergic signalling mediated both by D1 receptors and by D2 receptors. PMID- 10383651 TI - Nitric oxide is required for expression of LTP that is induced by stimulation phase-locked with theta rhythm. PMID- 10383650 TI - Interocular temporal delay sensitivity in the visual cortex of the awake monkey. AB - Due to the separation of the eyes, temporal retinal disparities are created during binocular stimulation and they have been proposed to be the basis of several stereo-visual effects. This paper studies the sensitivity of cortical neurons from area V1 to interocular temporal delay in the awake monkey (Macaca mulatta). Forty-four cells were included in this study. Temporal delay sensitivity was observed in 59% of them. About half of these temporal-delay sensitive cells were also sensitive to the stimulation sequence of the eyes. The cells that preferred one eye to be stimulated first were termed asymmetrical (46%); those which were not sensitive to the eye sequence of stimulation were termed symmetrical (54%). No clear differences were observed in the distribution of delay-sensitive cells according to their eye dominance. Fifty-six percent of balanced cells and 65% of unbalanced cells were sensitive to interocular delay. These data underline the importance of temporal cues for depth perception. PMID- 10383652 TI - Sexual imprinting, learning and speciation AB - Learned mate preferences may play an important role in speciation. Sexual imprinting is a process whereby mate preferences are affected by learning at a very young age, usually using a parent as the model. We suggest that while the origins of learning appear to lie in the advantages of individual recognition, sexual imprinting results from selection for recognition of conspecifics. This is because efficient early learning about one's own species is favoured in the presence of heterospecifics. If different species are hybridizing, both sexual imprinting and learning to avoid heterospecifics during adulthood promote assortative mating and hence speciation. As a result of sexual imprinting, speciation may also be completed in allopatry when divergence between populations is sufficient to prevent interbreeding when the populations reunite, even in the absence of genetic evolution of mate preference. The role of behaviour and learning in completing the speciation process is relatively overlooked. In particular the evolution of sexual imprinting as a result of selection against hybridization warrants more study. PMID- 10383653 TI - Population structure and gene flow of the brazilian shrub helicteres brevispira AB - Helicteres brevispira is a pioneer species of the tropical riparian forest whose populations appear to cycle through episodes of extinction and recolonization. Therefore, genetic consequences of founding events may strongly affect the genetic structure of its populations. An analysis of F-statistics showed that the studied population of H. brevispira is genetically substructured with the highest values of FST found in areas of high plant densities. Spatial autocorrelation analysis showed that genetic patches have diameters of 3-6 m. Although pollinator movements are usually between plants which are 3-6 m apart, longer flights occur and the neighbourhood area is estimated to have a diameter of 15 m. This suggests that genetic patches are smaller than the neighbourhood area. Seed dispersal is limited, mostly less than 2 m from the mother plants. Thus, short seed dispersal, seed dormancy and founder effects in the seed bank may be the most important determinants of genetic structure in populations of H. brevispira. Factors such as drift and inbreeding may also increase the level of substructure in this population, but the equilibrium model of isolation by distance does not fit our data. PMID- 10383654 TI - Genomic coalescence in a population of laxmannia sessiliflora (Angiospermae, anthericaceae): an association of lethal polymorphism, self-pollination and chromosome number reduction AB - A population of Laxmannia R. Br. (Angiospermae, Anthericaceae) near Collie, Western Australia, combines the taxonomically significant sessile inflorescences of L. sessiliflora Dcne. (n = 4) and the derived breeding system of L. ramosa Lindl. (n = 4). It exhibits a polymorphism for seed-aborting lethal equivalents, significant levels of self-pollination and a chromosome polymorphism in which a haploid genome with n = 3 is most frequent. Allozyme analysis indicates that the population is either of hybrid origin or one that has uniquely diverged from a phylogenetic link between the two species. The population is considered to represent a natural demonstration of the phenomenon of genomic coalescence as modelled by James (1992, Heredity, 68, 449-456) in which devices which reduce the number of independently segregating supergenes heterozygous for recessive lethals are elevated to high frequencies by inbreeding. The population also suggests a mechanism whereby dysploid chromosome number reduction may be promoted by natural selection in natural population systems. PMID- 10383655 TI - A test of the hypothesis of an autopolyploid vs. allopolyploid origin for a tetraploid lineage: application to the genus barbus (Cyprinidae) AB - A new method is described for determination of the origin of polyploid lineages. It tests the hypothesis that a tetraploid lineage originated via autopolyploidization vs. allopolyploidization. The method is based on the hypothesis that, in the case of autopolyploidy, any genetic marker in the first tetraploid ancestor is represented by two copies (one for each homoeologous chromosome of the haploid complement), whereas in allopolyploidy some markers absent from one of the hybridizing species will display one copy at most. The model requires knowledge of the phylogeny (topology and branch lengths) of a sample of species descending from the same tetraploidization event, together with the number of homoeologous copies present in each species for a set of neutral markers. The likelihood of a given proportion of the markers being present in both homoeologous chromosome pairs of the ancestral tetraploid is expressed as a function of the deletion rate of a marker. In the case of an autopolyploid origin, this proportion equals one. A likelihood-ratio test was carried out to test this hypothesis. The method was used to examine five microsatellite loci in eight species of Barbus (sensu lato). Assuming the validity of the hypotheses on phylogenetic relationships and evolutionary rates, the test rejects the possibility that European tetraploid barbs originated through autopolyploidy. This is the first test that can reject autopolyploidy, and it would appear particularly useful for phylogenetic studies in taxa where hybridization is known and where, consequently, undetected reticulate evolution may impair phylogenetic reconstruction. PMID- 10383656 TI - Population cytotype structure in the polyploid galax urceolata (Diapensiaceae) AB - The geographical distributions of diploid and polyploid Galax urceolata overlap in the Blue Ridge Mountains, USA. As part of an investigation into the evolutionary forces governing the establishment of polyploids and their coexistence with diploids, we examined the population frequencies of diploids, triploids and tetraploids in the area of overlap. Ploidy was inferred from estimates of DNA content, using flow cytometry, for 1570 individuals sampled from 42 populations. Across the entire sampling area, diploids and tetraploids were most abundant (55% and 34% of individuals, respectively), whereas triploids were least abundant (11%). Cytotype frequencies differed significantly among the northern, central and southern regions of the range (G = 649.02, d.f. = 4, P < 0.0001), with diploids most frequent in the north-east and least frequent in the south-west. Twenty-six per cent of the populations contained three cytotypes, 33% contained two, and 40% had a single cytotype. Populations with two cytotypes occurred in all possible cytotype combinations, but when triploids were present, they were always in the minority. Uniform populations were either diploid (81%) or tetraploid (19%), but never triploid. Overall, populations are predominantly diploid or tetraploid but rarely evenly mixed, suggesting disruptive selection for chromosome number in G. urceolata. The contribution of ecological sorting and frequency-dependent mating success to the distribution of polyploids and diploids is discussed. PMID- 10383657 TI - Genetic structure in three haploid peat mosses (Sphagnum) AB - Over the past 20 years, studies have revealed levels of genetic variation in bryophytes that are similar to those found in vascular plants. This has led many to question the traditional view of bryophyte evolution, which holds that these organisms have a low evolutionary rate. RAPD and isozyme analyses were used to measure genetic variation in 18 populations of several Sphagnum taxa, with special emphasis on the bisexual S. lindbergii and the unisexual S. angustifolium, S. fallax and S. isoviitae. Both types of markers were found to be selectively neutral. A test of population differentiation showed no significant divergence between S. fallax and S. isoviitae growing in sympatry; these taxa were therefore treated as conspecific. Only S. angustifolium had polymorphic isozyme loci. The highest genetic variation in RAPD loci was found in S. angustifolium; the lowest in S. lindbergii. There seemed to be a high turnover rate of individuals in S. angustifolium populations. Populations of S. fallax coll. were strongly differentiated for RAPD markers, whereas S. angustifolium populations were only weakly differentiated for any marker, even for populations from different continents. Populations of S. lindbergii were not differentiated at all. Most studied populations did not fit the 'Conocephalum - Plagiomnium' model of bryophyte population structure. The observed patterns could best be explained by assuming a low evolutionary rate, at least in S. angustifolium, meaning that high levels of molecular variability seem not to be incompatible with slow evolution. PMID- 10383658 TI - A theoretical basis for measuring the efficiency of selection in plant breeding AB - A new index of selection efficiency, i.e. the ratio of the probability of achieving the desired genetic gain in a target population to the cost expended for that gain, is introduced to define the optimum selection procedures that give the most opportunities for producing new commercial varieties. Monte Carlo simulations of mass selection showed that the index gives solutions to some important optimization problems which cannot be solved based on the traditional indices, the expected genetic gain or its ratio to the cost. The optimum cycle and intensity of selection and optimum population size for a target population could be defined by the new index, but not by the traditional indices. PMID- 10383660 TI - The role of ecophysiological models in QTL analysis: the example of specific leaf area in barley AB - Crop modelling has so far contributed little to the genetic analysis of a quantitative trait. This study illustrates how a simple model for crop phenological development, which assumes that crop development rate is affected by daily effective temperature, can assist the identification of Quantitative Trait Loci (QTLs), using specific leaf area (SLA) in barley as an example. The SLA was measured in a field experiment six times during the growing season of 94 recombinant inbred lines (RILs) derived from a cross between cultivars Prisma and Apex. Of the six measurements, one was conducted at the same physiological age for all RILs (at flowering), four were undertaken at specific chronological days prior to flowering, and the last one was taken at 14 days after flowering. When the measured SLA was directly used as the quantitative trait, one to three QTLs were detected for SLA at each measurement time. The major dwarfing gene denso segregating in the population was found to affect SLA strongly at all measurement times except at flowering. If SLA of the different RILs was corrected for differences in physiological age at the time of measurement, by the use of the crop development model, QTLs were detected for SLA at only three stages. Furthermore, the effect of the denso gene was no longer significant during the preflowering stages. The effect of the denso gene detected in the first instance was therefore the consequence of its direct effect on the duration of the preflowering period. This demonstrates the important role that crop development models can play in QTL analysis of a trait that varies with developmental stage. Potential uses of ecophysiological crop growth models in QTL analysis are briefly discussed. PMID- 10383659 TI - Cloning and physical mapping of Atlantic salmon (Salmo salar L.) telomeric sequences. AB - We cloned and sequenced DNA containing telomeric sequences from Atlantic salmon (Salmo salar L.). At both low and high stringency conditions, a cloned probe containing (TTAGGG)n sequences binds only at the extreme termini of the chromosomes, not at interstitial sites. Blocks of interstitial telomere-like sequences were expected from the chromosome polymorphism because of Robertsonian translocations that exist in Atlantic salmon. Our results support the hypothesis that telomeric sequences were almost entirely lost during chromosome fusion events. PMID- 10383661 TI - Spatial and temporal genetic structure in chloroplast and allozyme markers in phacelia dubia implicate genetic drift AB - For neutral genes, uniparental inheritance is expected to reduce effective population size relative to biparentally inherited genes. In finite populations, the ensuing genetic drift can cause stronger spatial and temporal differentiation. An intrapopulation polymorphism in chloroplast DNA was used to examine relative spatial and temporal population structure of chloroplast and allozyme markers in the annual plant Phacelia dubia. There was significant differentiation among populations at chloroplast markers but not for allozyme loci. A fine-scale analysis showed significant structure among sites within populations for chloroplast markers and local heterozygote deficiencies at allozyme loci. These spatial analyses suggest that gene flow via pollen exceeds that via seed. Temporal variation in chloroplast markers, assessed over a 10-year period, was evident in two of four populations, and allozyme loci were characterized by temporal variation in rare-allele frequencies. Population structure appeared to be related to the intensity and type of human disturbance influencing each population. Habitat destruction promoted isolation and enhanced differentiation, whereas mowing increased seed dispersal and reduced differentiation for chloroplast markers. At this time, genetic drift appears to be the primary force shaping chloroplast gene frequencies. PMID- 10383662 TI - Patterns of allozymic variation within calluna vulgaris populations at seed bank and adult stages AB - We investigated the spatial genetic structure within and between two plots of Calluna vulgaris and the extent to which the soil seed bank differed genetically from adults at seven allozyme loci. Averaged over the two plots, the seed bank and adult populations contained very similar levels of genetic diversity. Moreover, seeds contained in a single soil core (100 cm3) exhibited similar mean allozyme diversity to the surrounding adult population, indicating that the seed bank preserves genetic diversity at a very local scale. Few differences in allelic frequencies were found between the seed bank and its surrounding adult population in each plot. Mean GST indicated a lack of differentiation between the two plots at adult (GST = 0.008) and seed bank (GST = 0.002) stages. Low interplot differentiation is consistent with the outcrossing mating system of the population (tm = 0.91 in one plot) and its history of human disturbance. In contrast, spatial autocorrelation analysis of adults indicated a genetic structure at a very local scale, with positive autocorrelation for all alleles below 2 m in one plot and with a pattern of positive autocorrelation below 8 m in the two plots. Current limitation to seed dispersal rather than spatial extension of clones is thought to be responsible for local genetic structure. PMID- 10383663 TI - Factors influencing the extent of inbreeding depression: an example from scots pine AB - Detailed studies suggest that the level of inbreeding depression may vary between populations. In a study of Scots pine from Finland, the level of inbreeding depression was much lower in northern than in southern populations. We have examined theoretically whether population genetic factors, such as the level of selfing, intensity of selection against heterozygotes or homozygotes, level of mutation, a bottleneck, finite population size, or the level of polyembryony could account for this difference. Higher selfing or stronger selection against heterozygotes in the north, both at biologically reasonable levels, appear to produce changes consistent with the observed differences and we consider these to be the most likely explanations. In addition, the differences could have accumulated by these mechanisms over the age of the northern population, approximately 100 generations. Finally, the differences generated by these factors could still be maintained in the face of reasonable levels of gene flow from the south. Such a comprehensive theoretical investigation of this example has given some general insight into the potential influence of these evolutionary factors on the level of inbreeding depression and provides an approach that could be used to understand similar phenomena in other examples. PMID- 10383664 TI - Effect of insect-mediated dispersal on the genetic structure of postglacial water mite populations AB - Assaying population structure in species that differ in dispersal ability can help to determine whether population differentiation is dependent on the movement of individuals between populations. Here, allozyme variation is analysed in over 1100 individuals from nine species and two species complexes of Arrenurus water mites collected throughout north-eastern North America. As larvae, eight taxa are obligate parasites of winged adult insects that provide the primary opportunity for dispersal. Three additional species have lost the ability to parasitize insects and do not disperse in this manner. Consistent with the glaciated history of the region, very low allozyme heterozygosity was found in these taxa (Ho = 0.00-0.12), near panmixia in five out of seven species for which population differentiation was calculated and no patterns of isolation by distance over spatial scales up to several hundred kilometres. Nonetheless, in two out of three comparisons between sister species with and without parasitic larvae, parasitism was significantly associated with higher heterozygosity. Population differentiation could also be contrasted for two of these sister species pairs; in each case, lower estimates of FST were found in the mites able to disperse on insects. The statistical significance of these contrasts was dependent on the method used to estimate variance. At the scale of the genus, behavioural differences among insect vectors allows for broader hypotheses that relate water mite genetic diversity to dispersal ability. For the genus, rank correlations of dispersal ability with direct count heterozygosity (n = 11) and population differentiation (n = 7) were not significantly different from zero. These results are consistent with the hypothesis that allozyme population structure is primarily the result of historical patterns in these regions. However, comparisons between sister species suggest a limited role for dispersal in homogenizing populations genetically, even when drift-gene flow equilibrium has not been achieved. PMID- 10383665 TI - Genetic differentiation and natural hybridization between two morphological forms of the common woodlouse, oniscus asellus linnaeus 1758 AB - The common woodlouse Oniscus asellus can be divided into two forms on the basis of morphology, particularly male accessory genitalia. Where these taxa meet, morphological intermediates are found, and the forms were therefore described as subspecies; O. a. asellus and O. a. occidentalis. In this study allozyme loci are used to test the hypothesis that intermediate forms result from hybridization, and to study the nature of hybridization. Thirteen enzyme loci were scored across five English sites representative of each subspecies and intermediates. Ten loci showed strong frequency differences between asellus and occidentalis populations, although no loci showed completely fixed differences. These data confirm that asellus and occidentalis represent genetically distinct taxa, and that intermediate populations are of hybrid origin. There is apparently substantial population substructuring in the contact zone, as indicated by deficits of heterozygotes (FIS) and sporadic gametic (i. e. linkage) disequilibria. Population structure in the Oniscus hybrid zone appears to be analogous to that seen in plant hybrid swarms rather than the narrow hybrid zones observed in many animal taxa. Values of Nei's genetic distance between the subspecies range from 0.65 to 0.70; these are much higher than between typical conspecific taxa and are indicative of ancient genetic divergence. However, because asellus and occidentalis do not remain distinct in areas of overlap, it is simplest to regard these taxa as members of the same species. PMID- 10383666 TI - Testing kin selection with sex allocation data in eusocial hymenoptera AB - Sex allocation data in eusocial Hymenoptera (ants, bees and wasps) provide an excellent opportunity to assess the effectiveness of kin selection, because queens and workers differ in their relatedness to females and males. The first studies on sex allocation in eusocial Hymenoptera compared population sex investment ratios across species. Female-biased investment in monogyne (= with single-queen colonies) populations of ants suggested that workers manipulate sex allocation according to their higher relatedness to females than males (relatedness asymmetry). However, several factors may confound these comparisons across species. First, variation in relatedness asymmetry is typically associated with major changes in breeding system and life history that may also affect sex allocation. Secondly, the relative cost of females and males is difficult to estimate across sexually dimorphic taxa, such as ants. Thirdly, each species in the comparison may not represent an independent data point, because of phylogenetic relationships among species. Recently, stronger evidence that workers control sex allocation has been provided by intraspecific studies of sex ratio variation across colonies. In several species of eusocial Hymenoptera, colonies with high relatedness asymmetry produced mostly females, in contrast to colonies with low relatedness asymmetry which produced mostly males. Additional signs of worker control were found by investigating proximate mechanisms of sex ratio manipulation in ants and wasps. However, worker control is not always effective, and further manipulative experiments will be needed to disentangle the multiple evolutionary factors and processes affecting sex allocation in eusocial Hymenoptera. PMID- 10383667 TI - Population structure and genetic variation of european wild rabbits (Oryctolagus cuniculus) in east anglia AB - The European wild rabbit (Oryctolagus cuniculus) is an introduced species in Britain, and populations have been profoundly influenced by both man and disease. In stable environmental conditions, distinct social behaviour is observed, and this social structure leads to significant genetic structuring at the intrapopulation level. In this study, European wild rabbits were sampled from 17 sites across the East Anglian region of Britain and genotyped with nine microsatellite loci. Genotypical proportions deviated significantly from Hardy Weinberg equilibrium, reflecting a degree of population subdivision and non random mating. Several estimates of measures of population genetic structure revealed that populations are genetically distinct and have small effective population sizes. These distinctive properties are seen to be the combined effects of the social structure and random drift acting on bottlenecked populations after myxomatosis. It is concluded that the genetic structure seen in rabbit populations today is unlikely to reflect historical structuring present before myxomatosis, but that it results from recent events. PMID- 10383668 TI - An evaluation of the use of pooled samples in studies of genetic variation. AB - When using molecular markers to study genetic variation, either the sampled individuals can be analysed individually or the individuals can be pooled and only the pools analysed (pooled samples). A theoretical investigation was carried out into the use of pooled samples in the detection of alleles and providing maximum likelihood estimates of allele frequency. The results show that, in many cases, pooled samples are more efficient than samples of individuals. Of the different pool sizes studied, small pools containing two or three individuals showed the smallest expected squared error of allele frequency estimates. PMID- 10383669 TI - The colonization history and present-day population structure of the european great tit (Parus major major) AB - The colonization history and present-day population structure of the European subspecies of the great tit Parus major major were studied using mitochondrial control region sequences. One major haplotype was found in all but one of the eight sampled populations from Spain to northern Finland. The other haplotypes differed from the common one by just a few substitutions; the overall nucleotide diversity was 0.00187 and haplotype diversity 0.8633. No population structuring was detected. The mismatch distribution followed the expected distribution of an expanding population. The estimated time to the most recent common ancestor coincides with the last glacial period. The results suggest that P. m. major survived the last glacial period in a single isolated refuge probably by the Mediterranean Sea. This was followed by rapid colonization of the European continent and population growth. The most recent range expansion northwards is still occurring. Gene flow between the sampled populations is extensive. It is aided by juvenile dispersal, long-distance movements of juvenile flocks and partial migration in the northern parts of the great tit's range. PMID- 10383670 TI - Genetic differentiation and phylogenetic relationships among greek silurus glanis and silurus aristotelis (Pisces, siluridae) populations, assessed by PCR-RFLP analysis of mitochondrial DNA segments AB - Mitochondrial DNA diversity of seven Silurus glanis populations (six from Greece and one from the Danube Delta) and three populations of the endemic Greek Silurus aristotelis was investigated. RFLP analysis of four regions of mitochondrial DNA (cytochrome b, D-loop, ND-5/6) amplified by PCR was used. Ten and nine haplotypes were found in S. glanis and S. aristotelis, respectively. No haplotype was shared between the two species. Significant geographical substructuring was observed in the distribution of haplotypes, with most populations possessing private haplotypes. These haplotypes can serve as genetic 'tags' and therefore warrant protection. Haplotype diversity was very low for all Greek S. glanis populations, possibly because the small size and large annual fluctuations of Greek inland waters do not support large fish populations. Nucleotide divergence was in the range of 0.00-0.52% among S. glanis populations, and 0. 00-0.11% among S. aristotelis populations. Historical factors such as glaciations could account for these low values. The value of 6. 75% sequence divergence of the two species refutes the classification of the two species in different genera, as proposed by some authors. This study constitutes the first attempt, based on mitochondrial molecular data, to address the complicated evolutionary history of the two species which belong to the widely distributed and economically important Siluridae family. PMID- 10383671 TI - An approach for predicting heterosis based on an additive, dominance and additive x additive model with environment interaction. AB - A method is proposed for predicting potential heterosis of offspring of crop hybrids by an additive, dominance and additive x additive model (ADAA). By using unbiased predictors of additive and dominance, as well as additive x additive effects, general formulae for predicting heterosis over mid-parent and heterosis over the better parent are derived for different generations. When there exists genotype by environment (GE) interaction, formulae are also derived for predicting interaction heterosis. Heterosis in a specific environment is the sum of heterosis arising from the main genotypic effect and that arising from GE interaction deviation. The epistasis heterosis (DeltaAA) could play an important role in the use of heterosis for both an F1 hybrid and its later generations. In addition, a simple formula is given for predicting the number of generations of a cross that would still keep a certain level of heterosis over the better parent. Data from a diallel cross of cotton are analysed as a worked example for predicting genotypic value, heterosis, and the number of generations for each cross when heterosis over the better parent is larger than 5%. PMID- 10383672 TI - Effect of population size on the mating system in a self-compatible, autogamous plant, aquilegia canadensis (Ranunculaceae) AB - In self-compatible plants, small populations may experience reduced outcrossing owing to decreased pollinator visitation and mate availability. We examined the relation between outcrossing and population size in eastern Ontario populations of Aquilegia canadensis. Experimental pollinations showed that the species is highly self-compatible, and can achieve full seed-set in the absence of pollinators via automatic self-pollination. We estimated levels of outcrossing (t) and parental inbreeding coefficients (F) from allozyme variation in naturally pollinated seed families for 10 populations ranging in size from 32 to 750 reproductive individuals. The proportion of seeds produced through outcrossing was generally low (mean = 0.29 +/- 0.02 SE) and varied widely among populations (range = 0.00-0.83). Accordingly, estimates of F were large (mean = 0.26 +/- 0.05) and significantly greater than zero in seven populations. As expected, four small populations (N < 40) outcrossed less (0.17 +/- 0.03) than six large populations (N > 90; 0.38 +/- 0. 03). However, parental plants were not significantly more inbred in small than large populations (P = 0.18). There was no difference in the germination of seeds from hand self- and cross-pollinations. However, population genetic estimates of inbreeding depression for survival expressed from seed to reproductive maturity were very high (mean delta = 1 - relative fitness of selfed seed = 0.88 +/- 0.14). The combination of self compatibility and automatic self-pollination makes the mating system of A. canadensis sensitive to variation in ecological factors that affect the likelihood of cross-pollination. PMID- 10383673 TI - Properties and natural occurrence of maternal-effect selfish genes ('Medea' factors) in the red flour beetle, tribolium castaneum AB - Maternally acting selfish genes, termed 'Medea' factors, were found to be widespread in wild populations of Tribolium castaneum collected in Europe, North and South America, Africa and south-east Asia, but were rare or absent in populations from Australia and the Indian subcontinent. We detected at least four distinct genetic loci in at least two different linkage groups that exhibit the Medea pattern of differential mortality of genotypes within maternal families. Although each M factor tested had similar properties of maternal lethality to larvae and zygotic self-rescue, M factors representing distinct loci did not show cross-rescue. Alleles at two of these loci, M1 and M4, were by far the most prevalent, M4 being the predominant type. M2 and M3 were each found only once, in Pakistan and Japan, respectively. Although M1 could be genetically segregated from M4 and maintained as a purified stock, the M1 factor invariably co-occurred with M4 in field populations, whereas M4 usually occurred in the absence of other Medea factors. The dominant maternal lethal action of M1 could be selectively inactivated (reverted) by gene-knockout gamma irradiation with retention of zygotic rescue activity. PMID- 10383674 TI - Heritability of tibia fluctuating asymmetry and developmental instability in the winter moth (Operophtera brumata L.) (Lepidoptera, geometridae) AB - Broad-sense heritability of fluctuating asymmetry and developmental instability in the winter moth were analysed in a full-sib breeding experiment. Effects of both genetic background and common environment on both tibia FA (measured for the three pairs of legs) and body size were studied. As body size has previously been shown to be a reliable indicator of larval feeding success and expected fitness, the relationship between FA and body size was investigated as well. This relationship is of interest because it has been argued that the low heritability of FA results from a strong relationship between FA and fitness. Broad-sense h2 of body size equalled zero whereas the effect of common environment was strong. The heritability of FA was low and not statistically significant for separate tibias. For FA based on the average of the three tibias h2 equalled 0.07 and differed significantly from zero. The heritability of developmental instability equalled 0.09. Thus the use of the hypothetical repeatability to translate the h2 of FA to h2 of developmental instability did not result in a strong increase in this species. Individual asymmetry was not correlated with fitness (as estimated by body size), indicating that the low heritabilities of FA are not a consequence of a strong correlation with fitness. Between-trait correlations in the unsigned FA were significant. However, these correlations are not necessarily indicative of an individual asymmetry parameter as the signed FA values were positively correlated as well, suggesting interdependent development of the three pairs of legs. Further research is necessary to investigate what the effects of interdependent development are on patterns in FA. PMID- 10383675 TI - Inbreeding depression and partial selfing: evolutionary implications of mixed mating in a coastal endemic, silene douglasii var. oraria (Caryophyllaceae) AB - Recent studies have found moderate to high levels of selfing in plants despite high inbreeding depression. Because both factors influence the evolution and persistence of rare plants, we conducted glasshouse and field studies of pollination and inbreeding in Silene douglasii var. oraria, a perennial tetraploid endemic to coastal prairies. We detected: (i) variation in reproduction or inbreeding depression among life stages, years and maternal families; (ii) partial selfing yet higher relative fitness in outcrossed than selfed progeny; (iii) differing values of selfing and inbreeding depression using population means vs. matched maternal families. Fruit and seed production varied significantly with pollination treatment and year in flowers manipulated in situ during three seasons of growth. Hand-pollinations providing pollen in excess of ovule production in 1996 yielded more seeds than marked, open-pollinated flowers, implying pollen limitation of seed production. However, among-year differences in reproductive success after open-pollination (i.e. values equivalent to autogamy, selfing or outcrossing) suggest that pollination levels also vary temporally. In pollinations matched by maternal family, selfing yielded significantly fewer seeds than outcrossing. Fitness differences between inbred and outbred progeny were significant (P < 0.05) for seed production, percentage germination, and biomass or fecundity, but not for survival. Maternal family data gave selfing rates intermediate between obligate outcrossers and predominant selfers (S = 0.34 0.51), but population-wide means gave unusually high values (S = 1.1-1.6). Cumulative inbreeding depression was 76% for maternal families, and 70-85% using population means; in all cases, inbreeding depression values were high in early and late life stages, and lowest for survival. Thus far, reproductive assurance offers the most cogent explanation for the coexistence of moderate selfing and high inbreeding depression in this strongly protandrous Silene once thought to be highly outcrossing. This possibility merits further study in other rare plants with mixed-mating systems, where inbreeding depression and pollinator scarcity may both compromise population persistence and raise the threshold below which selfing is favoured by evolution. PMID- 10383676 TI - Genetic analysis of temperature-dependent transmission of mitochondrial DNA in Drosophila. AB - In artificially induced mitochondrial DNA (mtDNA) heteroplasmy in Drosophila, the effects of chromosome substitution on temperature-dependent selection in mtDNA transmission were investigated. Using two strains of D. melanogaster, bw;e11 and y;bw;st, which showed a different temperature dependency in mtDNA transmission, chromosomes were substituted reciprocally, and mtDNA of D. mauritiana was introduced into each newly constructed strain. For each heteroplasmy, the transmission of mtDNA was examined at 25 degrees C and 19 degrees C. When either the second or the third chromosome of the y;bw;st strain was substituted with that of the bw;e11 strain, the temperature-dependent selection in mtDNA transmission was altered. The selection was not changed when either the second or the third chromosome of the bw;e11 strain was substituted with that of the y;bw;st strain, or even when both the second and the third chromosomes of the bw;e11 strain were substituted with those of the y;bw;st strain. These results suggest that the temperature-dependent selection in mtDNA transmission is co operatively regulated by gene products that are encoded by the X, second and third chromosomes. PMID- 10383677 TI - Spatial autocorrelation analysis of individual multiallele and multilocus genetic structure. AB - Population genetic theory predicts that plant populations will exhibit internal spatial autocorrelation when propagule flow is restricted, but as an empirical reality, spatial structure is rarely consistent across loci or sites, and is generally weak. A lack of sensitivity in the statistical procedures may explain the discrepancy. Most work to date, based on allozymes, has involved pattern analysis for individual alleles, but new PCR-based genetic markers are coming into vogue, with vastly increased numbers of alleles. The field is badly in need of an explicitly multivariate approach to autocorrelation analysis, and our purpose here is to introduce a new approach that is applicable to multiallelic codominant, multilocus arrays. The procedure treats the genetic data set as a whole, strengthening the spatial signal and reducing the stochastic (allele-to allele, and locus-to-locus) noise. We (i) develop a very general multivariate method, based on genetic distance methods, (ii) illustrate it for multiallelic codominant loci, and (iii) provide nonparametric permutational testing procedures for the full correlogram. We illustrate the new method with an example data set from the orchid Caladenia tentaculata, for which we show (iv) how the multivariate treatment compares with the single-allele treatment, (v) that intermediate frequency alleles from highly polymorphic loci perform well and rare alleles poorly, (vi) that a multilocus treatment provides clearer answers than separate single-locus treatments, and (vii) that weighting alleles differentially improves our resolution minimally. The results, though specific to Caladenia, offer encouragement for wider application. PMID- 10383678 TI - History can be more important than 'pollination syndrome' in determining the genetic structure of plant populations: the case of aconitum lycoctonum (Ranunculaceae) AB - The Aconitum lycoctonum complex is a widespread yellow-flowered group of species found in central and southern Europe. Because of extreme morphological variability, the systematics of this group is confusing, and hybridizations among taxa are often hypothesized. To determine whether hybridization, realized mating system within populations or colonization from different Pleistocene refugia might explain some of the morphological variation, the genetic structure of 19 populations from central and southern Europe was studied using starch gel electrophoresis and 10 enzyme loci, of which eight were polymorphic. A pattern typical of an outcrossing species was expected because A. lycoctonum flowers are adapted to pollination by long-tongued bumblebees. However, heterozygosity was very low (between 0.031 and 0.150), which is atypical of either widespread or outcrossing species. The inbreeding coefficient, FIS, also suggested inbreeding in more than half the populations. Analysis of molecular variance showed that 31% of the genetic variation is found among populations, again suggesting inbreeding. A neighbour-joining tree based on Reynolds's genetic distance showed a clear separation between the central/eastern European populations and the samples from the Iberian peninsula and Alpes Maritimes. These data are consistent with the hypothesized refugia and the phylogeographical histories of several European forest trees. The genetic identity of all populations was very high, suggesting that all investigated populations belong to the same species despite high morphological variability. Hybridization was not supported by the data. PMID- 10383679 TI - The evolutionary genetics of ageing and longevity. AB - Evolutionary theories of ageing are based on the observation that the efficacy of natural selection decreases with age. This is because, even without ageing, individuals will die of environmental causes, such as predation, disease and accidents. Ageing is thought to have evolved as the result of optimising fitness early in life. A second process, namely the progressive accumulation of mutations with effects late in life, will reinforce this result. Longevity of a species is therefore determined by the amount of environmental mortality caused by the ecology of a species. The experimental data concerning the relative roles of both processes are reviewed here. Recent discoveries of the levelling of mortality curves, and of age specific mutations in mutation accumulation lines of Drosophila melanogaster, require adjustments to the original models of the evolution of ageing and species longevity. These adjustments do not invalidate the underlying rationale of evolutionary theories of ageing. With current developments in QTL mapping and genetic association studies, the unravelling of the ageing process has the potential to progress rapidly. PMID- 10383680 TI - Selection under negative linkage disequilibrium. Random mating versus inbreeding. AB - A population resulting from the diallel cross of a set of elite inbred lines was selected under two strategies: (i) inbred strategy (IS) under brother-sister mating; and (ii) random strategy (RIS) under random mating. The final lines in both cases were completely inbred. RIS populations clearly responded better than IS populations. Although most of the RIS advantage was achieved in the second generation, the average response under the RIS strategy increased with respect to that of the IS strategy from the beginning to the end of the experiment. In general terms and on theoretical grounds this RIS advantage is expected only if the initial disequilibrium is negative. One of the final inbred lines performed significantly better than the sum of the two inbred ancestors, strongly suggesting a heterotic epistatic combination fixed in homozygosity. Strong negative disequilibrium and partial epistatic control of quantitative characters are to be expected in breeding programmes of self-pollinating crops. In all likelihood, recurrent selection under forced random mating in such crops would result in better responses. PMID- 10383681 TI - Low genetic diversity among pea aphid (Acyrthosiphon pisum) biotypes of different plant affiliation. AB - Genetic diversity in the pea aphid Acyrthosiphon pisum was investigated by a restriction fragment length polymorphism (RFLP) analysis of three maternally inherited genomes (mitochondrial DNA and plasmids of the symbiotic bacteria Buchnera). Twenty-nine parthenogenetic clones of three A. pisum biotypes, defined by their capacity to use the legume crops pea, alfalfa and red clover, respectively, were analysed, and a total of 67 restriction sites was scored. No restriction site variation in the mitochondrial genome was obtained, but length variation at two regions (the A + T-rich region and ND3-ND5 region) was noted. One aphid clone bore a variant HindIII restriction site in the Buchnera leucine plasmid (pAPEleu), and two clones were heteroplasmic for a 0.76-kb deletion in the Buchnera tryptophan plasmid (pAPEtrp). Based on arthropod nucleotide substitution rates, it is proposed that the crop-feeding biotypes of A. pisum may have diversified within the last 100 000 years and possibly much more recently, since the advent of agriculture. PMID- 10383682 TI - Discriminating among cattle breeds using genetic markers. AB - Genetic markers provide a potentially powerful means of identifying the breed of individual animals. In this study diallelic and microsatellite loci were compared for their efficiency in discriminating among cattle breeds. Data were simulated for seven European cattle breeds using allele frequencies estimated at 20 microsatellite and 30 diallelic markers. Animals were assigned to the breed for which their genotype had the highest probability, and the power of the method assessed by estimating the error rate or proportion of animals misclassified. The number of markers required for discriminating among pure, or both pure and crossbreed, animals was investigated using either randomly sampled markers or markers selected on individual error rate. The relationship between individual marker variability and discriminatory power was also investigated. Microsatellite markers were found to be more powerful than diallelic markers for distinguishing among the breeds. The most discriminatory markers were those with the highest average heterozygosity and observed number of alleles. The number of markers needed to achieve a particular error rate could be reduced by selecting markers with the lowest individual error rates. Discrimination among both crossbreeds and pure breeds required approximately three times as many markers as discrimination among pure breeds alone. PMID- 10383684 TI - Fine-scale genetic structure and clinal variation in silene acaulis despite high gene flow AB - We investigated whether the distribution of genes reflects the patchy distribution of individuals of Silene acaulis on Pennsylvania Mountain in central Colorado. Five polymorphic protein loci were analysed using both F-statistics and spatial autocorrelation. Low thetaPOP (FST) indicated little genetic differentiation between populations approximately 1 km apart. This indicates high gene flow within our study site, perhaps as a result of long-distance pollen dispersal. Despite little differentiation between populations, there was clinal variation at the 6-Pgd-1 locus and significant within-population genetic structure (indicated by both thetaPATCH and spatial autocorrelation). We infer that this fine-scale genetic structure is the result of limited seed dispersal combined with genetic drift. The level of genetic structure varied markedly among populations, with the greatest genetic structure (highest Moran's I and thetaPATCH values) in two low-altitude, small, low-density populations. Intensive sampling such as used in this study may reveal similar patterns of fine-scale genetic differentiation in other patchily distributed plant species, particularly those with limited seed dispersal. PMID- 10383683 TI - A stable triple Wolbachia infection in Drosophila with nearly additive incompatibility effects. AB - Drosophila simulans strains infected with three different Wolbachia strains were generated by experimental injection of a third symbiont into a naturally double infected strain. This transfer led to a substantial increase in total Wolbachia density in the host strain. Each of the three symbionts was stably transmitted in the presence of the other two. Triple-infected males were incompatible with double-infected females. No evidence was obtained for interference between modification effects of the different Wolbachia strains in males. Some incompatibility was observed between triple-infected males and females. However, this incompatibility reaction is not a specific property of triple-infected flies, because it was also observed in double-infected strains. PMID- 10383685 TI - RAPD variation within and between natural populations of the wild rice oryza rufipogon from china and brazil AB - Genetic variation within and between eight natural populations of Oryza rufipogon from China and Brazil was investigated at the DNA level by analysis of RAPD fragments. Out of 60 random primers, which were initially screened against DNA from four individuals, 20 generated highly reproducible RAPD fragments which were then used for further population analysis. With these primers, 95 discernible DNA fragments were produced and 78 (82.1%) were polymorphic, which indicated that high levels of genetic variation existed in these natural populations. In addition, the Chinese populations showed greater polymorphism than those from Brazil at both the population and regional levels. This is noteworthy considering that the Chinese populations are from a relatively restricted area of China. The factors responsible for these findings include the contrasting mating systems in the Brazilian and Chinese populations, and gene flow from annual cultivated rice to perennial natural populations in China. An Analysis of Molecular Variance (AMOVA) was used to apportion the variation between individuals within populations, between populations within regions, and between regions. Results showed that 61.8% of the total genetic diversity resided between the two continents, whereas only 14.9% and 23.3% was attributable to population differences within regions and to individual differences within a population, respectively. The great genetic differentiation between the Chinese and Brazilian populations is in agreement with recent treatment of the American form of O. rufipogon as a separate species, O. glumaepatula. PMID- 10383686 TI - Microsatellite polymorphism and genetic impact of restocking in mediterranean brown trout (Salmo trutta L.) AB - The genetic impact of restocking Mediterranean brown trout populations with hatchery stocks was investigated in the Orb River drainage (France), using genetic data from three microsatellite loci. We sampled two wild populations, the main river which is restocked each year and one of its tributaries which has not been restocked for 6 years. Each sample was divided into two age groups (juveniles/adults). Introgression of each native population by hatchery stocks was previously estimated using allele frequencies from two diagnostic protein coding loci and one mtDNA haplotype. The genetic structure and allelic frequency at three microsatellite loci in native populations were compared with two hatchery samples belonging to stocks usually used for restocking this drainage. High levels of polymorphism (23-27 alleles per locus) were detected for two loci, whereas the third was less polymorphic. Polymorphism was significantly higher in the restocked population than in the now undisturbed population. Significant differences between age groups were observed in the main river, but not in its tributary. The introgression estimates using microsatellites were compared to those obtained from proteins and mtDNA. The different possible origins of alleles common to hatcheries and wild populations (homoplasy, ancestral polymorphism or introgression) are discussed. PMID- 10383687 TI - Allozyme variation and genetic structure of calluna vulgaris (heather) populations in scotland: the effect of postglacial recolonization AB - Recent fragmentation of populations as well as historical postglacial recolonization may have significantly affected the population genetic diversity of temperate plant species. Regional allozymic variability was measured at seven loci within and among 12 populations of Calluna vulgaris in the previously glaciated region of Scotland. These results were compared with existing data on south-western continental populations. Low genetic differentiation (FST = 0.024) and lack of consistent geographical pattern were found at the regional level among Scottish populations, implying a high rate of gene flow (Nm = 10.2), probably favoured by the nearly continuous range of C. vulgaris across Scotland and characteristics of the Scottish environment. Scottish populations possessed lower mean allozymic diversity (PLP = 40.48, A = 1.95, He = 0.133) than populations from all the continental regions investigated previously. Belgian populations were genetically more closely related to Scottish than to other continental populations. These last two findings are interpreted with regard to the evolutionary history of the species revealed by palynological data. PMID- 10383688 TI - The influence of triploidy on gene expression in the silkworm, bombyx mori AB - In Bombyx mori, it is well established that polyploids are easily induced when newly laid eggs are exposed to a variety of conditions, such as high or low temperature, centrifugal force, or chemicals like colchicine. To investigate gene dosage effects by varying the ploidy, the transcription levels of six genes expressed in various tissues were analysed in the diploid and two different genetically produced triploids (PPC and CCP). In the PPC triploid, the transcription level per cell of two genes was directly proportional to the structural gene dosage, whereas two other genes showed the mRNA level expected if compensation occurred. In the CCP triploid, three genes displayed dose-dependent levels of expression, whereas one gene showed the same expression level as the diploid strains. In both triploids, exceptional cases showed a negative correlation of expression with ploidy or a positive correlation greater than expected from the structural gene dosage. Interestingly, the transcription levels of most tested genes were significantly different from the strains which were used as parents of the triploids, and also widely divergent expression patterns were found for some genes in the diploid offspring. In this study, the cause of the unexpected expression patterns observed in the euploid series is discussed in relation to the difference between the two parental strains in expression level of genes and in trans-acting regulatory effects on their target genes. PMID- 10383689 TI - Mapping quantitative trait loci for complex binary traits in outbred populations. AB - Complex binary traits have a dichotomous phenotypic expression but do not show a simple Mendelian segregation ratio. These traits are considered to be jointly controlled by the actions of several genes and a random environmental effect. The binary phenotype and the underlying factor are assumed to be linked through a threshold model. The underlying factor, referred to as the liability, is treated as a regular but unobservable quantitative character. Mapping quantitative trait loci (QTL) can be performed directly on the liability. Methods of QTL mapping for the liability of a complex binary trait have been well developed in line-crossing experiments. However, such a method is not available in outbred populations which usually consist of many independent pedigrees (families). In this study, we develop a method to analyse jointly multiple families of an outbred population. The method is developed based on a fixed-model approach, i.e. the QTL effects, rather than the variance, are estimated and tested. After the test, the estimated effects are then converted into a single estimate of the QTL variance by taking into consideration errors in the estimated effects. The QTL effects and variance covariance matrix of the estimates are obtained by a fast Fisher-scoring method. Monte Carlo simulations show that the method is not only powerful but also generates very accurate estimates of QTL variances. PMID- 10383690 TI - Group founding and breeding structure in the subsocial spider stegodyphus lineatus (Eresidae) AB - Co-operative behaviour may evolve by enhancing the genetic similarity of group members. Increased group similarity is thought to be the basis for the 'subsocial route' of social evolution in the spider family Eresidae. Two processes may promote the similarity of individuals within populations or breeding groups, namely philopatry in stable environments and founder events in a stochastic environment. We show that both processes led to genetic differentiation within and among populations of the subsocial spider Stegodyphus lineatus. Within populations we distinguished between the genetic structure caused by random mating and philopatry in old breeding groups and that caused by newly founded groups consisting of sibs. Such sib-groups suggest that new breeding groups are established primarily by single females. The different gene coancestries among breeding groups resulted in high variances among single-locus data. The results imply that sex-specific dispersal behaviour (random male mating-dispersal or female group founding) had different impacts on the population structure. This type of population structure, where within-population philopatry and founder events may lead to differential proliferation of breeding groups, is very similar to that presumed for social spiders, and is also one that could provide the conditions for interdemic selection. PMID- 10383691 TI - Errors in histopathology reporting: detection and avoidance. AB - The histopathological diagnosis is the bedrock of modern oncology, and plays a major role in the treatment of many other types of disease. Errors in these reports can critically affect patient care and may become the subject of media concern. This article considers how audit in histopathology can provide information about errors and inconsistencies in the diagnosis of surgical specimens. The use of audit to generate information about the background level of errors in pathology reports is reviewed, along with findings about the nature of these errors and the types of specimens more commonly affected. Generic audit strategies that can be used to minimize the risk of errors in reports are discussed, together with the use of audit to evaluate diagnostic criteria and pathological scoring or grading systems. The role of audit in determining the informational content of reports is included, and there is consideration of the relationship between sample size and error rates. The limited extent to which audit can be used to assess the performance of individual pathologists is also covered. PMID- 10383692 TI - Malignant phyllodes tumours of the breast display increased stromal p53 protein expression. AB - AIMS: To determine the variation in p53 protein expression in phyllodes tumours and fibroadenomas of the breast. METHODS AND RESULTS: Fifteen phyllodes tumours (six malignant, nine benign) and 20 fibroadenomas were examined for p53 expression by immunohistochemistry. Five of the six malignant phyllodes tumours showed moderate or strong p53 positivity at sites of peri-epithelial stromal condensation and atypia. All 20 fibroadenomas, nine benign phyllodes tumours and one malignant phyllodes tumour showed either negativity or focal weak nuclear positivity of scattered stromal cells. CONCLUSIONS: Increased p53 immunoreactivity is present in malignant phyllodes tumours in contrast to benign phyllodes tumours and fibroadenomas. Malignant phyllodes tumours display a distinctive pattern of p53 immunostaining which may be of diagnostic value. These findings suggest that p53 protein may be important in the progression of benign to malignant phyllodes tumours. PMID- 10383693 TI - p53 but not bcl-2 is expressed by most cholangiocarcinomas: a study of 28 cases. AB - AIMS: To examine the frequency and pattern of expression of p53 and bcl-2 in archival material from patients with cholangiocarcinomas and to evaluate their respective roles in its pathogenesis, diagnosis and prognosis. METHODS AND RESULTS: Twenty-eight surgical cases of cholangiocarcinomas diagnosed at St James's University Hospital and 16 control cases were immunostained with monoclonal antibodies to p53 and bcl-2 using streptavidin-biotin complex method. Pressure cooker was used for antigen retrieval. Of the cholangiocarcinomas, 85.7% (24/28) overexpressed p53. The intensity of staining in these cases varied from 1+ in 2, 2+ in 10 and 3+ in 12 cases. None of the 28 tumours expressed bcl-2. The well differentiated nature of the tumour made assessment of dysplasia difficult, however, where present it did not express p53 or bcl-2. The bile duct epithelium adjacent to the tumour and in the control cases did not show any significant nuclear staining for either antigen. CONCLUSIONS: Overexpression of p53 appears to play an important role as a late event in the pathogenesis of cholangiocarcinomas, while we found no evidence of bcl-2 overexpression. The expression of p53 in 86% of the invasive tumours, as compared to its lack in the adjacent normal bile duct epithelium, makes it potentially useful in the diagnostic histopathology of these cases. PMID- 10383694 TI - Low rate of apoptosis and overexpression of bcl-2 in Epstein-Barr virus associated gastric carcinoma. AB - AIMS: Epstein-Barr virus (EBV) has been demonstrated in about 10% of gastric carcinomas. However, the pathogenetic role of EBV in gastric carcinoma is uncertain. We compared the rate of apoptotic cell death, cell proliferation and the expression of apoptosis-related proteins in gastric carcinomas with or without EBV. METHODS AND RESULTS: Epstein-Barr virus was detected in 40 gastric carcinomas by EBV-encoded small RNA-1 in-situ hybridization. Apoptotic cell death, MIB-1, p53, bcl-2 and bcl-x were examined by the terminal deoxynucleotidyl mediated dUTP-nick end labelling method and immunohistochemistry. We also included 40 age-, sex- and disease stage-matched EBV-negative cases as a control. The number of apoptotic cells was significantly lower in EBV-positive (20 +/- 15. 1/1000 cells) and bcl-2-positive (17 +/- 12.9/1000 cells) tumours than in EBV negative (43 +/- 37.1) and bcl-2-negative tumours (38 +/- 32.1, P < 0.001, P < 0.001, respectively). bcl-2 immunostaining was significantly higher in EBV positive tumours (24 cases) than in EBV-negative tumours (12 cases, P < 0.05). There was no significant difference in bcl-x and p53 expression between EBV positive and -negative tumours. The number of MIB-1-positive cells in EBV positive tumours (237 +/- 161/1000) was significantly lower than in EBV-negative tumours (480 +/- 208/1000 cells, P < 0.001). CONCLUSIONS: A low rate of apoptosis and high bcl-2 expression were recognized in EBV-positive gastric carcinomas, suggesting that bcl-2 protein is the main inhibitor of apoptosis in EBV-positive carcinomas. In addition, the low apoptotic and proliferative activities may reflect a low biological activity in EBV-positive gastric carcinomas. PMID- 10383695 TI - Tumour responding accessory cells in testicular seminoma: an immunohistochemical study. AB - AIM: To investigate the role of accessory cells (and other chronic inflammatory cells) in the host immune response to testicular seminoma by defining their immunophenotypic characteristics and topographical arrangement. METHODS AND RESULTS: A panel of antibodies applicable to paraffin-embedded tissues was employed to characterize the host chronic inflammatory response in eight cases of classical testicular seminoma. The antibodies were directed against CD45RO, CD20, CD68, acid cysteine proteinase inhibitor (ACPI), MAC387, muramidase (MUR), S100 protein, Factor XIIIa, CD21 and HLA Class II. In all cases the majority of the inflammatory cells were T-lymphocytes situated mainly in areas of apparent tumour destruction. Large numbers of macrophages/dendritic cells which had not been evident by conventional light microscopy were also demonstrated. In particular, an immunophenotypically distinct population of accessory cells showing a specific pattern of distribution was revealed. It clearly rimmed islands of tumour and showed strong positive staining for CD68, MAC387 and HLA Class II. CONCLUSION: The study has identified an immunophenotypically distinct population of accessory cells showing a characteristic topographical arrangement. It is proposed that it represents a subpopulation of macrophages which are responding directly to the tumour and are likely to play a part in influencing tumour dynamics. PMID- 10383696 TI - Melan-A/Mart-1 expression in various melanocytic lesions and in non-melanocytic soft tissue tumours. AB - AIMS: The purpose of this study was to test different malignant non-melanocytic tumours with the commercially available antibody Melan-A to examine its diagnostic specificity and to compare the S100, Melan-A and HMB-45 reactivity in various melanocytic lesions. METHODS AND RESULTS: Seventy-three benign and malignant melanocytic lesions and 31 cases of non-melanocytic tumours, sarcomas, carcinomas and carcinoids, were selected. Immunohistochemical staining of paraffin sections, following a high temperature antigen unmasking technique, was performed. Melan-A stains junctional and dermal melanocytes in all benign melanocytic lesions with the exception of neuro-naevoid areas. The epithelioid and the spindle cells in malignant melanomas did not show considerable difference in their Melan-A reactivity. The predominantly spindle cell type mucosal melanomas contained more Melan-A-positive cells than HMB-45-positive cells and similar results were observed in metastatic malignant melanomas. In desmoplastic melanomas the positivity of Melan-A was not consistent. None of the sarcomas, carcinomas and carcinoids expressed Melan-A. Almost all soft tissue tumours, except for two malignant gastrointestinal stromal tumours, were unreactive for HMB-45. These two cases did not react with Melan-A antibody. CONCLUSIONS: Melan-A is a useful additional marker to differentiate non-melanocytic tumours from primary or metastatic melanoma. In melanocytic lesions the Melan-A staining pattern is similar to S100, but seems to be more specific. In desmoplastic melanomas, however, the variable Melan-A staining further necessitated detailed histological examination and the use of the S100 reaction. PMID- 10383697 TI - Combined clear and granular cell leiomyoma of soft tissue: evidence of transformation to a histiocytic phenotype. AB - AIMS: We present an unusual case of leiomyoma with a clear and granular cell pattern in which there was immunohistochemical evidence of transformation to a histiocytic phenotype. METHODS AND RESULTS: A 64-year-old man presented with mild scrotal swelling and pain. A local excision was performed after the clinical diagnosis of epidermal inclusion cyst. In the pathological specimen, another tumour nodule was identified which was composed predominantly of clear cells, with an occasional mixture of granular cells. Immunohistochemical analysis demonstrated positive staining for vimentin, lysozyme, CD68 and HAM56, but complete negativity for desmin, alpha-smooth muscle actin, HHF35, S100 protein, neurone-specific enolase and CD34. Ultrastructural study revealed dilated rough endoplasmic reticulum, glycogen granules, abundant vacuolar structures and also thin microfilaments with subplasmalemmal dense bodies. CONCLUSIONS: Based on these findings, we have interpreted it to be a rare case of leiomyoma with extensive clear cell and granular cell degeneration (combined clear and granular cell leiomyoma). This complete transformation of the immunohistochemical profile into the histiocytic phenotype has not been previously described in the literature. PMID- 10383698 TI - Leiomyoma with atypical cells (atypical leiomyoma) in the larynx. AB - AIMS: To report and confirm the identity of laryngeal leiomyoma with many atypical cells, which has not been described previously. CASE DETAILS: A 53-year old man was found to have a polypoid tumour in the larynx. The tumour was excised and the patient has shown no evidence of recurrence over a 5-year period. The tumour tissue comprised intersecting fascicles of spindle-shaped tumour cells with blunt-ended nuclei. Many of the tumour cells showed marked atypia. Mitotic activity in the tumour cells was low, and no atypical mitoses were found. Immunohistochemically, the tumour cells were positive for smooth muscle actin and desmin. p53 overexpression was identified in many tumour cells; the p53 labelling index of the tumour cells was 45%. DNA from tumour cells showed loss of heterozygosity on chromosomes 3p, 5q, 8p, 9p, 10q, 17p and 18q. We diagnosed this case as leiomyoma with atypical cells (atypical leiomyoma) based on the clinical course and pathological and genetic findings. CONCLUSION: This is the first report of atypical leiomyoma in the larynx. The clinical course and pathological findings indicate that although laryngeal atypical leiomyoma contains numerous atypical cells, it is a benign neoplasm. PMID- 10383699 TI - Origin of hyperplastic epithelial cells in idiopathic collapsing glomerulopathy. AB - AIMS: Glomerular epithelial cell hypertrophy and hyperplasia are listed as the primary criteria for the diagnosis of collapsing glomerulopathy (CG), a distinct variant of focal segmental glomerulosclerosis. However, the extent of podocyte phenotypic alterations that occur in CG, and the origin of the hyperplastic epithelial cells remain to be established. METHODS AND RESULTS: Renal biopsy materials from seven out of three patients with CG were studied by serial section analysis for immunohistochemistry and electron microscopy. Markers for podocytes (PHM5 and synaptopodin), parietal epithelial cells (PECs: cytokeratin) and macrophages (CD68) were used for the immunohistochemistry. Multiple ultrathin sections from a total of 15 glomeruli, including some from patients with CG, were examined by electron microscopy. Glomerular adhesions occurred in 71% of the serially sectioned glomeruli taken from patients with CG. Hyperplastic epithelial cells were immunonegative for podocyte markers and CD68, but invariably immunopositive for cytokeratin. Electron microscopy revealed that detachment of the podocytes from involved glomerular capillary walls was extensive. Many of the detached podocytes appeared to be necrotic and apoptotic. In contrast, junctional complexes of desmosomes and zonula adherens connected hyperplastic epithelial cells to each other. Cilia were also often observed. CONCLUSIONS: The results of our ultrastructural and immunohistochemical study suggest that the hyperplastic epithelial cells observed in cases of CG are derived from PECs. Our results raise the possibility that PECs play a general role in covering glomerular tufts from which the podocytes have disappeared. PMID- 10383700 TI - Herpetic salpingitis and fallopian tube prolapse. AB - AIM: We describe the unusual association of fallopian tubal prolapse and herpetic infection, an occurrence not previously reported to our knowledge. METHODS AND RESULTS: A 37-year-old woman presented with a small polypoid mass of the vaginal vault, 3 months after abdominal hysterectomy and abdominoplasty. The vaginal mass proved to be the fimbriated end of a fallopian tube, herniated into the vagina. Reintervention 3 months later with resection of a small vaginal 'polyp' revealed a residual portion of fallopian tube, with superimposed herpes simplex virus (HSV) infection and marked cytological atypia of surface epithelial cells. HSV-2 immunostaining of viral nuclear inclusions and of atypical cells confirmed the herpetic nature of the infection. CONCLUSION: Involvement of the genito-urinary tract by HSV may occur via an ascending infection from the cervix, but the fallopian tube, deeply located in the pelvis, is generally spared from herpetic infection. In the setting of fallopian tubal prolapse, direct exposure of the herniated fallopian tube to various pathogens in the vagina provides an unique clinical model for salpingitis. In herpetic tubal infections, special attention must be paid to cytological atypia of probable viral cytopathogenic origin, to avoid a misdiagnosis of malignancy. PMID- 10383701 TI - Redistribution of glomerular epithelial cell adhesion molecules in membranous glomerulonephritis. PMID- 10383702 TI - Penile neuromas with multiple Meissner's corpuscles. PMID- 10383703 TI - Extramedullary haemopoiesis in the cervix. PMID- 10383704 TI - Sarcomatous transformation of chromophobe cell renal carcinoma. PMID- 10383705 TI - TP53 mutation in mucinous carcinoma of the breast. PMID- 10383706 TI - Micropapillary transitional cell carcinoma of the urinary bladder. PMID- 10383707 TI - Seminal vesicle carcinoma. PMID- 10383708 TI - Spindle cell tumours of the breast: practical approach to diagnosis. AB - Spindle cell tumours of the breast are uncommon and often present diagnostic challenges. The most important is the sarcomatoid/metaplastic carcinoma, which has monophasic and biphasic variants. Each of these groups presents special diagnostic difficulties. In the monophasic variant the mesenchymal component predominates and the epithelial element forms a minor component often detected only after immunohistochemical study. The spindle cell areas may be bland and therefore under-diagnosed as nodular fasciitis or fibromatosis. Alternatively they may be highly malignant with a pattern that is misinterpreted as primary sarcoma of the breast. In the biphasic variant, the difficulty is in distinguishing between sarcomatoid carcinoma, myoepithelial carcinoma or malignant phyllodes tumour. Other spindle cell lesions of the breast include the various myofibroblastic tumours, the spindle cell variant of adenomyoepithelioma, the varied primary breast sarcomas, metastatic tumours with spindle cell morphology and, finally, the very rare follicular dendritic cell tumour. A simple practical approach to the diagnosis of spindle cell lesions is presented to help the general surgical pathologist to compile a differential diagnosis and to arrive at the correct conclusion PMID- 10383710 TI - Low-grade metaplastic carcinoma of the thymus. AB - AIMS: Five cases of a characteristic low-grade thymic epithelial tumour are described that we suggest calling metaplastic carcinoma of the thymus. METHODS AND RESULTS: The patients' ages ranged from 44 to 71 (mean 56.2) years. Four of the patients were male. Three of five tumours showed invasion into mediastinal fat or pleura but, otherwise, all were well circumscribed. No metastases were present. Histologically, the tumours showed a biphasic pattern with solid carcinomatous areas merging gradually with a spindle cell component. Lymphocytes were rare. Cytological atypia and mitotic activity were variable in the solid areas, but slight or absent in the spindle cell component. On immunohistochemistry, the tumours showed expression of cytokeratin, vimentin and/or epithelial membrane antigen, both in the carcinomatous and spindle cell components. In two cases, actin expression was also present in both components. In one case, chromogranin, S100 protein, glial fibrillary acidic protein and neuron-specific enolase were expressed in at least some cells of both components. None of the patients had myasthenia gravis. All patients are alive without evidence of recurrence or metastasis. CONCLUSION: Metaplastic carcinoma of the thymus is a distinct clinicopathological entity that should be distinguished from the usually benign medullary thymomas and from the clinically aggressive carcinosarcomas and sarcomatoid carcinomas. PMID- 10383709 TI - Reliability of intraoperative frozen section and imprint cytological investigation of sentinel lymph nodes in breast cancer. AB - AIMS: The sentinel lymph node procedure enables selective targeting of the first draining lymph node, where the initial metastases will form. A negative sentinel node (SN) predicts the absence of tumour metastases in the other regional lymph nodes with high accuracy. This means that in the case of a negative SN, regional lymph node dissection is no longer necessary. Besides saving costs, this will prevent many side-effects of lymph node dissection. The aim of this study was to evaluate the reliability of intraoperative cytological and frozen section investigation of the SN to detect metastases. This would allow the axillary lymph node dissection to be performed in the same session as the SN procedure and the excision of the primary tumour in case of a positive SN. METHODS AND RESULTS: Seventy-four SNs were detected by gamma probe detection of nanocolloid and visual localization of Patent Blue accumulations in 54 women with stage T1-2N0M0 invasive breast cancer. The identified SN were immediately investigated by frozen section and imprint cytological investigation. Diagnoses were confirmed on the paraffin material, and in case of negative frozen section and paraffin haematoxylin and eosin sections, skip sections and immunohistochemistry were performed. Thirty-one SNs (42%) contained metastases, of which 27 were detected by the frozen section procedure (sensitivity 87%). There were no false positives (specificity 100%). The sensitivity of the imprints was 62% with a specificity of 100%. When evaluating the data per patient, for the frozen section procedure the sensitivity was 91% and the specificity 100%, and for the imprints, the sensitivity was 63% and the specificity 100%. There were no SNs in which the imprints showed metastases and the frozen section did not. CONCLUSIONS: Intraoperative frozen section analysis is a reliable procedure by which a high percentage of sentinel lymph node metastases can be detected in breast cancer patients without false positive results. This allows the surgeon to perform an immediate axillary lymph node dissection in case of positive SNs. In up to 10% of cases, the final paraffin sections will reveal micrometastases that were not detected by the frozen section, and in these patients axillary lymph node dissection will have to be performed in a second session. The imprint method is significantly less sensitive than the frozen section but may be used as an alternative when frozen section is not possible. PMID- 10383711 TI - Acidic fibroblast growth factor is progressively increased in the development of oesophageal glandular dysplasia and adenocarcinoma. AB - AIMS: To determine by immunohistochemistry and amplification of cDNA the relationship between fibroblast growth factor (FGF) expression and progressive changes in Barrett's oesophagus associated with oesophageal adenocarcinoma (OA). The FGFs are potent mitogens that possess angiogenic properties and the capability to regulate growth and differentiation of various cell types. They have also been implicated in the development and progression of numerous solid tumours, including some carcinomas of the aerodigestive tract, such as nasopharyngeal carcinoma and pancreatic adenocarcinoma. MATERIAL AND RESULTS: We studied the expression of the two prototypic FGFs, acidic FGF (FGF-1) and basic FGF (FGF-2), in OA and OA precursor lesions, including intestinal metaplasia (IM), low-grade dysplasia (LGD) and high-grade dysplasia (HGD). Fresh tissue from 10 OAs and four associated HGDs was available for the determination of FGF-1 and FGF-2 mRNA expression accomplished by the PCR amplification of cDNA. Using immunohistochemistry, we studied the expression of the FGF-1 and FGF-2 proteins in archival, paraffin-embedded tissue that was available from 17 oesophageal resection specimens that included OAs and OA precursor lesions. As compared to gastric fundic mucosal controls, OAs and HGDs showed significantly enhanced expression of FGF-1 mRNA and protein. IMs and LGDs showed significantly lesser degrees of FGF-1 immunoreactivity that were not increased over controls. In contrast, both the overall percentage of FGF-2-reactive OAs and the overall FGF-2 protein expression, assessed using an immunoreactivity score, are comparable to FGF-2 expression in controls. CONCLUSIONS: It appears that FGF-2 is ubiquitously expressed in OA and in normal oesophageal and gastric mucosa while significant FGF-1 expression is essentially restricted to HGD and OA. Our data also suggest that FGF-1 is sequentially upregulated in the progression from metaplasia to dysplasia and adenocarcinoma. PMID- 10383712 TI - Histogenesis of gastric foveolar metaplasia following duodenal ulcer: a definite reparative lineage of Brunner's gland. AB - AIMS: We aimed to clarify the histogenesis of gastric metaplasia in the duodenal mucosa, particularly in association with a reparative lineage of Brunner's glands. METHODS AND RESULTS: Using immunohistochemical methods with recently developed antimucin monoclonal antibodies (mAbs) that distinguish foveolar and deep mucins of the gastric type, as well as mAb MIB-1, the histogenesis of gastric metaplasia was investigated in the duodenal wall of 20 surgically resected specimens. In duodenal ulcers extending into Brunner's glands with destruction of the muscularis mucosae, proliferating cells positive for MIB-1 were scattered in Brunner's glands. Interestingly, a group of proliferating cells was often seen next to the ulcerated surface. These cells were also positive for M1 (gastric-foveolar type mucin) but negative for M2 (deep gastric and Brunner glands' mucin). In regenerating ducts through granulation tissue, the proliferating cell zone was elongated, above which foveolar-type cells positive for M1 but negative for M2 were detected, indicating that the G-zone is newly established in Brunner's glands at the floor of an ulcer to produce gastric foveolar cells. Subsequently, an organoid growth of the normal stomach mucosa is completed in the duodenum. CONCLUSIONS: This study indicates a possible histogenetic pathway of gastric metaplasia in close association with a reparative lineage of Brunner's glands, suggesting that the occurrence of the gastric foveolar type epi-thelium is not a simple expansion of Brunner's duct but a true metaplasia. PMID- 10383713 TI - p53 immunoreactivity in endometrial metaplasia with dysfunctional uterine bleeding. AB - AIM: Overexpression of p53 has been reported in endometrial carcinomas, especially in uterine papillary serous carcinoma (UPSC), to correlate with worse prognosis. Endometrial metaplasia is commonly encountered in patients with dysfunctional uterine bleeding (DUB) and may on occasion be difficult to distinguish from atypical endometrial hyperplasia or carcinoma on biopsies. The present study was initiated in the belief that metaplastic tissue might not show overexpression of p53 and would thus help to distinguish it from carcinomas of non-endometrioid histology. METHODS AND RESULTS: Paraffin-embedded tissue of endometrial biopsies with papillary metaplasia (22 cases), tubal metaplasia (five cases) and eosinophilic meta-plasia (seven cases) from patients with DUB were immunostained for p53 immunoreactivity. No evidence of hyperplasia was noted in any of the cases selected for the study. Twenty-eight cases of UPSC were included for comparison. Our study showed p53 overexpression in 25 of 28 (89%) UPSC. Weak and heterogeneous p53 immunoreactivity was present in 10 of 22 (45%) papillary metaplasias, four of five (80%) tubal metaplasias and four of seven (57%) eosinophilic metaplasias. Follow-up of 16-45 (median 32) months was unremarkable except for one patient with eosinophilic metaplasia who had simple endometrial hyperplasia in subsequent biopsy. CONCLUSIONS: The presence of weak and heterogeneous p53 immunoreactivity in metaplastic endometrium is unexpected and might be a consequence of DNA damage. Intense, diffuse and homogeneous p53 staining favours carcinoma. PMID- 10383714 TI - Malignant myoepithelioma of the vulva resembling a rhabdoid tumour. AB - AIMS: We report an example of malignant myoepithelioma of the vulva, which has not been hitherto described. We discuss the differential diagnosis and briefly review the literature. METHODS AND RESULTS: The lesion was found in an 81-year old woman as an indolent 40 mm tumour. The neoplastic cells showed a myoid, spindled, epithelioid and plasmacytoid phenotype. Hyalinization of extracellular material and myxoid changes were present. There was a partly solid and microcystic pattern and a tight cohesiveness of cells was lacking. The circumscribed multinodular tumour somewhat resembled an extrarenal rhabdoid tumour, having large tumour cells with prominent nucleoli and large amounts of acidophilic cytoplasm. Immunohistochemically, the tumour cells were immunoreactive for cytokeratin, vimentin, muscle-specific actin, alpha-smooth muscle actin, and S100 protein, but not for desmin, epithelial membrane antigen, factor VIII-related antigen, CD34 and CD31. CONCLUSIONS: The histological and cytomorphological appearance of the tumour well as the immunohistochemical findings suggest the diagnosis of malignant myoepithelioma, possibly derived from minor vestibulary glands or ectopic breast tissue. Differential diagnoses are, in particular, extrarenal rhabdoid tumour and 'proximal type' epithelioid sarcoma. Differentiation is important, because the tumours show a different behaviour and prognosis. PMID- 10383715 TI - Down-regulation of p27Kip1 expression is correlated with increased cell proliferation but not expression of p21waf1 and p53, and human papillomavirus infection in benign and malignant tumours of sinonasal regions. AB - AIMS: Although p27Kip1(p27) is a cyclin-dependent kinase inhibitor and a contribution to tumorigenesis has been hypothesized, the possible role in tumours arising in the nasal and paranasal sinus regions is still unknown. METHODS AND RESULTS: Seventy-six sinonasal tumours, including 28 inverted papillomas (IPs) and 48 squamous cell carcinomas (SCCs), were immunohistochemically investigated, along with 46 exophytic papillomas (EPs) of upper respiratory tract and 34 samples of normal paranasal sinus epithelium. The results were also compared with expression of p21WAF1 (p21) and p53, cell proliferation assessed in terms of Ki67 labelling indices (LIs), and human papillomavirus (HPV) infection. The average p27 scores decreased from normal through to malignant lesions, while Ki67 LI scores showed a stepwise increase, the inverse correlation between scores for all categories being significant (r = - 0.639, P < 0. 0001). In the SCCs, p27 expression was significantly higher in keratinizing than nonkeratinizing type tumours (P < 0.05), while there was no association with p21 and p53 expression. Although HPV DNAs for type 16 and 18 were detected in two (7.4%) of 27 EPs, six (35.8%) of 28 IPs, and nine (28.1%) of 32 SCCs, no relation with p27 scores was evident. CONCLUSION: Loss of p27 expression correlates with increased cell proliferation in sinonasal tumours. Moreover, the expression appears to be associated with keratinization in SCCs of the paranasal sinus. These findings indicate that p27 expression may be a useful marker for the dysregulation of cell kinetics in these tumours. PMID- 10383716 TI - Cystic partially differentiated nephroblastoma in an adult: an immunohistochemical, lectin histochemical and ultrastructural study. AB - AIMS: Cystic partially differentiated nephroblastoma (CPDN) is an uncommon renal multicystic tumour, usually affecting early infants. To our knowledge, this report describes the first case of CPDN occurring in an adult. METHODS AND RESULTS: A 45-year-old man was found incidentally to have a left renal cystic tumour, measuring 20 mm in diameter, at the lower pole far from the pelvis. The tumour was composed of multilocular cystic spaces of variable size and intervening septa without solid nodular areas. The cysts were lined by a single layer of flattened, hobnail, or columnar epithelium. The septa were made of mesenchymal cells, which were admixed with small numbers of loosely aggregated blastemal cells, occasional tubular structures in various stages of development, and a few glomeruloid structures. The tumour cells had no anaplasia, and mitoses were rare. Immunohistochemical and lectin histochemical studies revealed that the cyst lining epithelium and the tubular structures in the septa expressed predominantly the markers for distal tubules and collecting ducts. Ultrastructurally, the cyst lining cells closely resembled collecting duct cells while some tubular structures showed an immature nephrogenic morphology. The patient was alive and well without evidence of recurrence 11 months after surgery. CONCLUSIONS: CPDN does occur in adults, as experienced in Wilms' tumour, though its incidence is extremely low. This study suggests that CPDN may show maturation intermediate between cystic nephroma and Wilms' tumour, even in adult cases. PMID- 10383717 TI - Polyester fibre prosthetic anterior cruciate ligament implant rupture: necrosis of ingrown connective tissue. AB - AIMS: To describe the histopathological and microanalytical features in seven cases of ruptured Apex(R) polyester (Terylene(R)) fibre anterior cruciate ligament prosthesis. METHODS AND RESULTS: Transmitted and polarized light microscopy was performed in all cases; one case was investigated by immunohistochemistry, transmission electron microscopy and scanning electron microscopy, with backscatter and X-ray detectors for elemental microanalysis. For comparison we also studied synovial biopsy material and unused polyester fibres. In the excised ligaments there was much ingrowth of fibrous tissue accompanying a florid giant cell reaction to the individual intact polyester fibres throughout the ligaments. Phagocytosis of particles of prosthesis-derived material was demonstrated and a striking finding was of necrosis of the ingrown connective tissue in the central portions of the ligaments. Hyalinized areas and 'neoligament growth' were less striking. A consistent finding in the polyester fibres was of small particles containing antimony, used as a catalyst in the manufacturing process. CONCLUSIONS: The pattern of reaction to the prosthetic material and the presence of necrosis differ from previous descriptions in animal and human explants of this and other prosthesis types. The mechanical effect of the necrosis is unlikely to be of significance with this ligament, which is load bearing ab initio. PMID- 10383718 TI - The challenge of sentinel lymph node biopsy. PMID- 10383719 TI - Diversion colitis: new light through old windows. PMID- 10383720 TI - H-cadherin expression in breast cancer. PMID- 10383721 TI - Plasmacytoid monocytes express IL3-receptor alpha and differentiate into dendritic cells. PMID- 10383722 TI - Clear cell epithelial cords in an ectopic hamartomatous thymoma. PMID- 10383723 TI - Hepatoid adenocarcinoma of the pancreas. PMID- 10383724 TI - Multiple hepatocellular adenomas associated with long-term carbamazepine. PMID- 10383726 TI - In this issue PMID- 10383725 TI - Solitary fibrous tumour of the pleura with t(4;15)(q13;q26) PMID- 10383728 TI - Profilaggrin requires both linker and filaggrin peptide sequences to form granules: implications for profilaggrin processing in vivo. AB - Filaggrin is an intermediate filament associated protein that aids the packing of keratin filaments during terminal differentiation of keratinocytes. Premature aggregation of keratin filaments is prevented by filaggrin expression as the inactive precursor, profilaggrin, which is localized in keratohyalin granules in vivo. Profilaggrin is phosphorylated and contains multiple filaggrin repeats separated by a hydrophobic linker peptide. We have previously shown that filaggrin constructs containing the linker, when transiently transfected into epithelial cells, lead to expression of a protein that resembles keratohyalin (Dale et al. J Invest Dermatol 108:179-187 1997). To characterize further the region(s) of the linker and/or filaggrin that are necessary for granule formation, we generated several mutant constructs from Flag-FG-1, and generated fusions of filaggrin with green fluorescent protein. We also subjected profilaggrin to protein phosphatase 2A treatment and measured its subsequent solubility. We found that granular morphology is not dependent on the linker or conserved phosphorylation sites, nor is solubility affected by protein phosphatase 2A treatment. Granule morphology was abrogated only in a truncated construct, which still contains the linker. A construct consisting of 16 amino acids of filaggrin fused to green fluorescent protein led to rounded and bizarrely shaped transfected cells with compact keratin filaments, suggesting that very little filaggrin sequence is required for keratin filament interaction. Radiolabeled filaggrin-green fluorescent protein constructs specifically bound keratin in overlay assays confirming that the observed cytoskeletal collapse is due to filaggrin-keratin interaction. Our findings indicate that profilaggrin must be extensively processed before it loses both its granule forming ability as well as its insolubility, suggesting that granule formation in vivo correlates with insolubility in vitro. Further, filaggrin retains its ability to bind keratin as it is degraded to smaller peptides. PMID- 10383729 TI - Enhanced expression of melanocortin-1 receptor (MC1-R) in normal human keratinocytes during differentiation: evidence for increased expression of POMC peptides near suprabasal layer of epidermis. AB - Immunohistochemical staining of human skin specimen showed the stronger localization of proopiomelanocortin peptides near the suprabasal layer of the epidermis, where keratinocytes are mostly differentiated. To test the possibilities of whether the production of proopiomelanocortin peptides or their receptor-binding activity or both is increased during differentiation of keratinocytes, we treated the cells in culture with Ca2+ to induce their differentiation. The production of proopiomelanocortin peptides and its gene expression were not induced significantly, but the binding ability of melanocortin receptor, as well as its gene expression were stimulated by Ca2+. Ultraviolet B irradiation, an inducer of differentiation, stimulated both proopiomelanocortin production and melanocortin receptor expression. These data show that normal human keratinocytes express melanocortin receptor similar to melanocytes, and that it is induced during differentiation. PMID- 10383727 TI - The Nf1 tumor suppressor regulates mouse skin wound healing, fibroblast proliferation, and collagen deposited by fibroblasts. AB - Neurofibromatosis type 1 patients develop peripheral nerve tumors (neurofibromas) composed mainly of Schwann cells and fibroblasts, in an abundant collagen matrix produced by fibroblasts. Trauma has been proposed to trigger neurofibroma formation. To test if loss of the neurofibromatosis type 1 gene (Nf1) compromises fibroblast function in vivo following trauma, skin wounding was performed in Nf1 knockout mice. The pattern and amount of collagen-rich granulation bed tissue, manufactured by fibroblasts, was grossly abnormal in 60% of Nf1+/- wounds. Nf1 mutant fibroblasts showed cell autonomous abnormalities in collagen deposition in vitro that were not mimicked by Ras activation in fibroblasts, even though some Nf1 effects are mediated through Ras. Nf1+/- skin wound fibroblasts also proliferated past the normal wound maturation phase; this in vivo effect was potentiated by muscle injury. In vitro, Nf1+/- fibroblasts showed higher proliferation in 10% serum than Nf1+/+ fibroblasts. Macrophage-conditioned media or epidermal growth factor potentiated Nf1+/- fibroblast proliferation in vitro, demonstrating abnormal response of mutant fibroblasts to wound cytokines. Thus Nf1 is a key regulator of fibroblast responses to injury, and Nf1 mutation in mouse fibroblasts causes abnormalities characteristic of human neurofibromas. PMID- 10383730 TI - Basal transepidermal water loss is increased in platelet-type 12-lipoxygenase deficient mice. AB - The roles of fatty acids in the skin have been under investigation since early reports of the phenotypic abnormalities of mice fed a diet deficient in essential fatty acids. Little is known about the functional significance of fatty acid metabolism by lipoxygenases in epidermis. Here, we have examined the role of platelet-type 12-lipoxygenase which converts arachidonic acid to the oxygenated metabolite 12-hydroperoxyeicosatetraenoic acid, in the skin using platelet-type 12-lipoxygenase-deficient mice generated by gene targeting. Platelet-type 12 lipoxygenase in wild-type mice was localized to the stratum granulosum by immunohistochemical analysis. Platelet-type 12-lipoxygenase-deficient mice lacked immunodetectable platelet-type 12-lipoxygenase in platelets and epidermis, appeared grossly normal, and exhibited an increase in basal transepidermal water loss without alteration in basal mitotic activity. Water loss and mitotic activity in mice with an acetone-disrupted membrane barrier were normal. No defect in ultrastructural properties or content of major fatty acids in dorsal skin or ear inflammation response was apparent in platelet-type 12-lipoxygenase deficient mice. These results indicate that the platelet-type 12-lipoxygenase pathway in mice is partly responsible for normal permeability barrier function but the mechanism awaits further elucidation. PMID- 10383731 TI - Fibrin microbeads (FMB) as biodegradable carriers for culturing cells and for accelerating wound healing. AB - We have developed biodegradable fibrin-derived microbeads as potent cell carriers. The fibrin-derived microbeads, 50-200 microm in diameter, were tested for their attachment to a wide range of cell types. Fibrin-derived microbeads were shown to be greatly haptotactic to cells (such as endothelial cells, smooth muscle cells and fibroblasts), which respond to fibrinogen in contrast to keratinocytes and different cell lines derived from leukocytic lineage. The cells on fibrin-derived microbeads could be maintained for more than 10 d and achieved a high density. 31P-nuclear magnetic resonance was employed to monitor phosphate metabolism in cells, with densities on the order of 100 million cells per g of fibrin-derived microbeads. The 31P-nuclear magnetic resonance adenosine triphosphate and phosphocreatine signals, equivalent to the signal obtained with perfused normal skin, indicated that metabolism of cells on fibrin-derived microbeads was responsive to oxygenation and nutrients. Light, fluorescent, and confocal laser microscopy revealed that the porous fibrin-derived microbeads accommodate up to 200-300 cells due to their high surface area which minimized contact inhibition. Cells could degrade the fibrin-derived microbeads and be transferred to seed culture flasks without trypsinization. In a pig skin wound healing model, fibrin-derived microbeads + fibroblasts were transplanted into full thickness punch wounds. This procedure was compared with other treatment modalities, such as the addition of human platelet-derived growth factor BB or fibrin-derived microbeads alone. By the third day after wounding, only the wounds in which fibroblasts on fibrin-derived microbeads were added showed significant formation of granulation tissue. Based on the above, we project many uses of our novel fibrin-derived microbead technology for cell culturing, wound healing and tissue engineering. PMID- 10383732 TI - Neutral endopeptidase expression and distribution in human skin and wounds. AB - Cutaneous sensory nerves mediate inflammation and wound healing by the release of neuropeptides such as substance P. Neutral endopeptidase is a cell surface enzyme that degrades substance P and thereby terminates its biologic actions. The distribution of neutral endopeptidase in normal skin and wounded human skin, however, has not been examined. The objectives of this study were to evaluate neutral endopeptidase expression in wounded and unwounded skin as well as in cells derived from human skin. Neutral endopeptidase was strikingly localized in normal skin by immunohistochemistry to keratinocytes of the epidermal basal layer, to hair follicles, eccrine and sebaceous glands as well as to endothelium of blood vessels and to large nerves. Standard incisional human wounds were studied at several time points between 1 h and 28 d after wounding. Staining for neutral endopeptidase was noted in the wound bed 6 h after wounding. In contrast to normal skin, staining of all the epidermal cell layers was noted in the migrating tongue of epithelium in l d wounds. Similar full-thickness staining was noted in 3 d and 7 d wounds in all layers of the new wound epithelium and in a "transition epithelium" near the wound edge. By 28 d post wounding neutral endopeptidase staining again was detected only in the basal layer of the epidermis. Neutral endopeptidase mRNA was detected in normal skin and wounds as well as cultured keratinocytes, fibroblasts and endothelial cells. Neutral endopeptidase enzymatic bioactivity was demonstrated in cultured keratinocytes. While it is known that several metalloproteinases important to tissue repair are produced by keratinocytes, this is the first evidence that keratinocytes produce neutral endopeptidase. Neutral endopeptidase may terminate the proinflammatory and mitogenic actions of neuropeptides in normal skin and wounds. PMID- 10383733 TI - Expression of differentiation markers during fetal skin development in humans: immunohistochemical studies on the precursor proteins forming the cornified cell envelope. AB - The cornified cell envelope is formed during the terminal differentiation of epidermis through cross-linking of specific proteins by transglutaminases. The specific arrangement of individual protein in the cornified cell envelope and participation of individual protein in the cornified cell envelope at different regions of skin, i.e., palm, foreskin, lips, etc. are not clearly understood. In order to understand the pattern and expression schedule of each individual precursor protein during the differentiation and formation of cornified cell envelope, the expression of precursor proteins in developing human fetal skins from the first to the third trimester were examined by immunohistochemical studies. Involucrin was found in the periderm and intermediate layer from 14 wk estimated gestational age, while loricrin and small proline-rich protein 1 were found in the periderm from 16 wk estimated gestational age. Filaggrin and trichohyalin that are absent in the adult cornified cell envelope were found in the granular and horny layers from 24 wk estimated gestational age. The precursor proteins except trichohyalin did not change their patterns after the onset of initial expression during development. Trichohyalin was transiently expressed in the granular and horny layers of the epidermis from 24 wk estimated gestational age with peak expression at 27 wk estimated gestational age, but was not detected in adult skin. In hair follicles, trichohyalin expression was stable without change from 20 wk estimated gestational age. These findings suggest that fetal skin may have different sets of barriers from the second trimester; the immature cornified cell envelope is formed in the early second trimester and the mature cornified cell envelope is formed in the late second or early third trimester when filaggrin and trichohyalin appear. PMID- 10383734 TI - Live confocal microscopy of oligonucleotide uptake by keratinocytes in human skin grafts on nude mice. AB - Anti-sense oligonucleotide uptake by keratinocytes in human skin grafts on athymic mice was examined using live confocal microscopy. Fluorescein isothiocyanate-labeled 15-mer C-5 propyne modified phosphorothioate anti-sense oligonucleotide (10-50 microM) was intradermally injected into normal human skin grafts on athymic mice, and the localization of the anti-sense oligonucleotide was assessed after 1-24 h postinjection. Anti-sense oligonucleotide was found to localize in the nuclei of basal and suprabasal keratinocytes after 1-2 h, and this localization was still observed after 24 h. This live in vivo observation of anti-sense oligonucleotide uptake in basal keratinocytes was confirmed using conventional fluorescence microscopy of fixed sections of skin grafts. Neither single nucleotides which were fluorescein isothiocyanate-labeled nor fluorescein isothiocyanate alone was able to penetrate into the nuclei of human skin graft keratinocytes after intradermal injection, and hence it is likely that the anti sense oligonucleotide was not degraded prior to intracellular localization. Topical administration of anti-sense oligonucleotide and anti-sense oligonucleotide-liposome complexes resulted primarily in localization in the stratum corneum of human skin grafts. When grafts were tape stripped prior to anti-sense oligonucleotide administration, however, as little as 5 microM anti sense oligonucleotide was required to observe nuclear anti-sense oligonucleotide accumulation. These results suggest that cutaneous anti-sense strategies can be tested using delivery via intradermal anti-sense oligonucleotide injection in human skin grafts on athymic mice, and that agents providing penetration of anti sense oligonucleotide across the stratum corneum are likely to be required for successful topical therapies. PMID- 10383735 TI - The human papillomavirus type 11 upstream regulatory region triggers hair follicle-specific gene expression in transgenic mice. AB - We have generated transgenic mice carrying the URR of the human papillomavirus type 11 ligated in front of the Escherichia coli beta-galactosidase coding region sequence. Using X-Gal staining to demonstrate beta-galactosidase production, we observed a hair-specific transcription of the reporter gene. This transcription was limited to the epithelial cells of the hair bulge region. The transgene was developmentally regulated, as no LacZ staining was demonstrated during embryogenesis and specific staining was first observed after birth. Surprisingly, dexamethasone and ultraviolet B, but not phorbol myristate acetate or progesterone treatment of the animals resulted in an increase in number and intensity of hair follicles expressing the reporter gene. PMID- 10383736 TI - Predominant expression of Fas (CD95) ligand in metastatic melanoma revealed by longitudinal analysis. AB - The expression of Fas ligand has recently been proposed as a novel tumor escape mechanism for melanoma. To establish the characteristics of Fas ligand expression during the course of melanoma progression we performed a longitudinal study analyzing primary tumors as well as subsequently evolving metastases. In primary melanoma Fas ligand was expressed in two of 20 lesions; this expression was weak and restricted to few parts of the tumors. The Fas ligand positive primary melanomas were rather thick, i.e., 8.5 and 3.8 mm, versus a median of 2.4 mm of the remaining tumors. In contrast, for metastatic melanoma Fas ligand expression was present in six of 11 cases investigated. The metastases of primary tumors displaying Fas ligand maintained its expression. As Fas ligand positive melanoma cells are capable of inducing apoptosis in susceptible cells, e.g., Fas positive tumor infiltrating lymphocytes, we tested for the presence of apoptotic cells in situ by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling. This analysis revealed that apoptotic cells were present within the Fas ligand positive tumors. The number of apoptotic cells, however, never exceeded 5% of the total cells. Thus, Fas ligand mediated apoptosis does not seem to be a major immune escape mechanism for melanoma but its expression correlates with the stage of melanoma. PMID- 10383737 TI - Periderm cells form cornified cell envelope in their regression process during human epidermal development. AB - Terminally differentiated stratified squamous epithelium forms a lining of the plasma membrane called the cornified cell envelope, a thick layer of several covalently cross-linked precursor proteins including involucrin, small proline rich proteins, and loricrin. Their cross-linking isodipeptide bonds are formed by epidermal transglutaminases 1-3. Material from lamellar granules is attached on the extracellular surface of corneocytes during the keratinization process. The formation of cornified cell envelope and sequential expression of major cornified cell envelope precursor proteins, transglutaminases, and 25 kDa lamellar granule associated protein were studied in human embryonic and fetal skin. Ultrastructurally, membrane thickening has already started in periderm cells of the two-layered epidermis and an electron-dense, thickened cell envelope similar to cornified cell envelope in adult epidermis is observed in periderm cells at the three-layered and later stages of skin development. In the two-layered epidermis (49-65 d estimated gestational age), immunoreactivities of involucrin, small proline-rich proteins, all the transglutaminases, and lamellar granule associated protein were present only in the periderm. In the three-layered epidermis and thereafter (66-160 d estimated gestational age), loricrin became positive in the periderm cells, transglutaminases extended to the entire epidermis, and lamellar granule-associated protein was detected in intermediate cells as well as periderm cells. Immunoelectron microscopy demonstrated that both major cornified cell envelope precursor proteins, involucrin and loricrin, were restricted to the cornified cell envelope in periderm cells at this stage of development. After 160 d estimated gestational age, the periderm had disappeared and cornified cell envelope proteins and lamellar granule-associated proteins were expressed in the spinous, granular, and cornified cells and transglutaminases were detected in the entire epidermis. These findings indicate that cornified cell envelope precursor proteins, transglutaminases, and lamellar granule-associated proteins are expressed in coordination in periderm cells during human epidermal development and suggest that periderm cells form cornified cell envelope in the process of regression. PMID- 10383738 TI - Human epidermal differentiation complex in a single 2.5 Mbp long continuum of overlapping DNA cloned in bacteria integrating physical and transcript maps. AB - Terminal differentiation of keratinocytes involves the sequential expression of several major proteins which can be identified in distinct cellular layers within the mammalian epidermis and are characteristic for the maturation state of the keratinocyte. Many of the corresponding genes are clustered in one specific human chromosomal region 1q21. It is rare in the genome to find in such close proximity the genes belonging to at least three structurally different families, yet sharing spatial and temporal expression specificity, as well as interdependent functional features. This DNA segment, termed the epidermal differentiation complex, contains 27 genes, 14 of which are specifically expressed during calcium dependent terminal differentiation of keratinocytes (the majority being structural protein precursors of the cornified envelope) and the other 13 belong to the S100 family of calcium binding proteins with possible signal transduction roles in the differentiation of epidermis and other tissues. In order to provide a bacterial clone resource that will enable further studies of genomic structure, transcriptional regulation, function and evolution of the epidermal differentiation complex, as well as the identification of novel genes, we have constructed a single 2.45 Mbp long continuum of genomic DNA cloned as 45 p1 artificial chromosomes, three bacterial artificial chromosomes, and 34 cosmid clones. The map encompasses all of the 27 genes so far assigned to the epidermal differentiation complex, and integrates the physical localization of these genes at a high resolution on a complete NotI and SalI, and a partial EcoRI restriction map. This map will be the starting resource for the large-scale genomic sequencing of this region by The Sanger Center, Hinxton, U.K. PMID- 10383739 TI - Serum xylosyltransferase: a new biochemical marker of the sclerotic process in systemic sclerosis. AB - UDP-D-xylose:proteoglycan core protein beta-D-xylosyltransferase (EC2.4.2.26) is the initial enzyme in the biosynthesis of chondroitin sulfate and dermatan sulfate proteoglycans in fibroblasts and chondrocytes. Secretion of xylosyltransferase into the extracellular space was determined in cultured human dermal fibroblasts. A more than 6-fold accumulation of xylosyltransferase activity in cell culture supernatant was observed (day 1, 0.6 microU per 106 cells; day 9, 4.1 microU per 106 cells); however, intracellular xylosyltransferase activity remained at a constant level (0.4 microU per 106 cells). Exposure of human chondrocytes to colchicine led to a 3-fold decreased level of xylosyltransferase and chondroitin-6-sulfate concentration in cell culture. Specific xylosyltransferase activity and chondroitin-6-sulfate concentration decreased in a concentration-dependent manner and in parallel in culture medium and accumulated 5-fold in cell lysates indicating that xylosyltransferase is secreted simultaneously into the extracellular space with chondroitin sulfate proteoglycans. Xylosyltransferase activities were determined in serum samples of 30 patients with systemic sclerosis. Xylosyltransferase activities in female (mean value 1.28 mU per liter, 90% range 1.10-1.55 mU per liter) and male patients (mean 1.39 mU per liter, 90% range 1.16-1. 57 mU per liter) with systemic sclerosis were significantly increased in comparison with blood donors of a corresponding age. Furthermore, xylosyltransferase activity was correlated with the clinical classification of systemic sclerosis. Female patients with diffuse cutaneous systemic sclerosis showed higher serum xylosyltransferase activities than patients with limited systemic sclerosis. These results confirm that the increase of proteoglycan biosynthesis in sclerotic processes of scleroderma is closely related to an elevated xylosyltransferase activity in blood and demonstrate the validity of xylosyltransferase as an additional diagnostic marker for determination of sclerotic activity in systemic sclerosis. PMID- 10383740 TI - Downregulation of endothelin B receptor in human melanoma cell lines parallel to differentiation genes. AB - Normal human melanocytes have been shown to respond to the signal peptide endothelin by increased proliferation and melanin formation. Contradictory findings, however, have been reported about which of the two endothelin receptors (EDNRA or EDNRB) is expressed in normal melanocytes and melanoma cells. Moreover it was not clear whether malignant cells differ from their normal precursors in this respect. Screening a melanocyte cDNA library for genes downregulated in melanomas identified clones specific for EDNRB. Northern blots proved that the corresponding mRNA is generally expressed in cultures of human cutaneous melanocytes and congenital melanocytic nevus cells. In 16 of 17 melanoma cell lines, however, the expression of EDNRB mRNA was strongly downregulated. EDNRA was only weakly expressed and detectable by northern blotting in 12 of 17 cultures of benign melanocytic cells and four of 17 melanoma cell lines. Nested reverse transcriptase-polymerase chain reaction proved several melanoma cell lines to be completely negative for EDNRA expression. Gene deletion as the cause of missing endothelin receptor expression was ruled out by genomic Southern blots. Receptor binding assays confirmed RNA data revealing 1.6 x 105 endothelin 1 binding sites per cell for a melanocyte culture and between 8.7 x 104 and 400 sites per cell for melanoma cell lines. Expression of pigmentation genes coding for tyrosinase, TRP-1 and TRP-2 correlated positively with that of EDNRB but negatively with EDNRA expression. EDNRB but not EDNRA expression is therefore typical for melanocytic cells, and downregulation of EDNRB seems to be an important characteristic of melanoma cells possibly related to malignancy or apoptosis. PMID- 10383741 TI - The effects of radicals compared with UVB as initiating species for the induction of chronic cutaneous photodamage. AB - There is substantial evidence that ultraviolet radiation induces the formation of reactive oxygen species which are implicated as toxic intermediates in the pathogenesis of photoaging. The aim of this study was to determine whether repeated topical treatment with benzoyl peroxide, a source of free radicals, produced the same cutaneous effects as chronic ultraviolet B radiation. Three concentrations of benzoyl peroxide (0.1, 1.5, 5.0% wt/wt) and three cumulative fluences of ultraviolet B radiation (0.9, 2.2, 5.1 J per cm2) used alone and in all combinations along with appropriate controls. Female SKH1 (hr/hr) albino hairless mice were treated 5 d per wk for 12 wk. Extracellular matrix molecules and histologic parameters were assessed. Ultraviolet B radiation induced a fluence-dependent and time-dependent increase in skin-fold thickness. Fluence dependence was seen for epidermal thickness, sunburn cell numbers, dermal thickness, glycosaminoglycan content, mast cell numbers, and skin-fold thickness. Benzoyl peroxide treatment alone caused less marked increases in epidermal and dermal measures compared with ultraviolet B under the conditions used. A benzoyl peroxide concentration-dependent increase was only observed for elastin content, although the highest concentration of benzoyl peroxide increased epidermal thickness and glycosaminoglycan content. A synergistic interaction between ultraviolet B and benzoyl peroxide was not found. These results indicate that repeated administration of benzoyl peroxide produces skin changes in the hairless mouse that qualitatively resemble those produced by ultraviolet B and suggest that common mechanisms may be involved. In addition, any potential synergistic effect of ultraviolet B and benzoyl peroxide was below the level of detection used in this study. PMID- 10383742 TI - Low-dose UVA and UVB have different time courses for suppression of contact hypersensitivity to a recall antigen in humans. AB - This study investigates the relative effects of low-dose solar-simulated ultraviolet, ultraviolet A, and ultraviolet B radiation on the elicitation of contact hypersensitivity to nickel in nickel-allergic volunteers. A xenon arc lamp with changeable filters was used to irradiate groups of volunteers daily, on separate areas of their lower backs, with both solar-simulated ultraviolet (ultraviolet B, ultraviolet AII + ultraviolet AI) and ultraviolet A (same ultraviolet AII content but twice the ultraviolet AI as the solar-simulated ultraviolet spectrum) for 1 and 2 d; 3, 4, and 5 d; and from 1 to 4 wk. A fourth group was irradiated for 1-5 d with the ultraviolet B component of solar simulated ultraviolet. Following the final irradiation in each group, nickel containing patches were applied to both ultraviolet-treated sites and adjacent, unirradiated control sites. Erythema caused by nickel contact hypersensitivity at each site was quantitated 72 h later with a reflectance erythema meter. By comparing the nickel reactions of irradiated and unirradiated skin, ultraviolet immunosuppression was assessed with the different spectra and durations of ultraviolet exposure. We found significant immunosuppression with daily doses of ultraviolet B and ultraviolet A equivalent to approximately 6 min of summer sun exposure, and that ultraviolet A and ultraviolet B exerted their maximal immunosuppressive effects at different times. Solar-simulated ultraviolet-induced immunosuppression was significant after one exposure, near-maximal after two exposures and remained elevated thereafter. Ultraviolet B-induced immunosuppression was lower than that induced by solar-simulated ultraviolet, but followed a similar time-course. In contrast, ultraviolet A-induced immunosuppression was transient, peaking after three exposures. Immune responses returned towards normal with subsequent ultraviolet A exposure, suggesting that an adaptive mechanism may prevent immunosuppression by continued ultraviolet A irradiation. PMID- 10383743 TI - Interferon-gamma is involved in photoimmunoprotection by UVA (320-400 nm) radiation in mice. AB - Ultraviolet B radiation not only inflicts tumor-initiating DNA damage, but also impairs T cell-mediated immunity relevant to survival of the initiated cells. We have reported, however, that ultraviolet A radiation, in contrast, is immunologically innocuous in hairless mice and opossums, but renders the animals resistant to the immunosuppression by ultraviolet B, or its mediator cis-urocanic acid. Ultraviolet B irradiation of skin causes abundant release of numerous cytokines (interleukin-1, interleukin-6, interleukin-10, tumor necrosis factor alpha); notably interleukin-12 and interferon-gamma do not appear to be upregulated. A recent report has indicated that interleukin-12 protects from photoimmunosuppression in mice, but it remains unclear whether interleukin-12 acts directly or via interferon-gamma, which it is known to stimulate. Here we investigate the possible role of interferon-gamma in UVA photoimmunoprotection, using interferon-gamma gene knockout mice in comparison with control C57/BL6 mice, and the systemic contact hypersensitivity reaction (induced by sensitization through a nonirradiated skin site) to measure immunity. interferon gamma-/- mice raised normal contact hypersensitivity responses, and were unaffected, as were C57BL control mice, by ultraviolet A exposure. In response to ultraviolet B irradiation or topical cis-urocanic acid treatment, control mice became immunosuppressed by 69% and 27%, respectively, and interferon-gamma-/- mice by 79% and 27%. When ultraviolet B exposure or cis-urocanic acid was followed by ultraviolet A irradiation, however, contact hypersensitivity was totally restored in control mice, but remained suppressed by 55% and 25%, respectively, in interferon-gamma-/- mice. Injection of recombinant interferon gamma in the interferon-gamma-/- mice restored the ultraviolet A protective effect against cis-urocanic acid-induced immunosuppression. These observations suggest that interferon-gamma plays a part in ultraviolet A immunoprotection from the suppressive effect of ultraviolet B radiation and, and that the mechanism appears to be via antagonism by this cytokine of the cis-urocanic acid immunosuppressive action. PMID- 10383744 TI - Infrared spectra of basal cell carcinomas are distinct from non-tumor-bearing skin components. AB - Infrared spectroscopy, by probing the molecular vibration of chemical bonds, directly indicates tissue biochemistry. An expanding body of literature suggests that infrared spectra distinguish diseased from normal tissue. The authors used infrared spectroscopy to examine basal cell carcinoma to explore distinctive characteristics of basal cell carcinoma versus normal skin samples and other skin neoplasms. Spectra of epidermis, tumor, follicle sheath, and dermis were acquired from unstained frozen sections, and analyzed qualitatively, by t-tests and by linear discriminant analyses. Dermal spectra were significantly different from the other skin components mainly due to absorptions from collagen in dermis. Spectra of normal epidermis and basal cell carcinoma were significantly different by virtue of subtle differences in protein structure and nucleic acid content. Linear discriminant analysis characterized spectra as arising from basal cell carcinoma, epidermis, or follicle sheath with 98.7% accuracy. Use of linear discriminant analysis accurately classified spectra as arising from epidermis overlying basal cell carcinoma versus epidermis overlying nontumor-bearing skin in 98.0% of cases. Spectra of basal cell carcinoma, squamous cell carcinoma, nevi, and malignant melanoma were qualitatively similar. Distinction of basal cell carcinoma, squamous cell carcinoma, and melanocytic lesions by linear discriminant analyses, however, was 93.5% accurate. Therefore, spectral separation of abnormal versus normal tissue was achieved with high sensitivity and specificity, which points to infrared spectroscopy as a potentially useful screening tool for cutaneous neoplasia. PMID- 10383745 TI - Expression and activation of matrix metalloproteinases in wounds: modulation by the tripeptide-copper complex glycyl-L-histidyl-L-lysine-Cu2+. AB - We investigated the expression and activation of matrix metalloproteinases in a model of experimental wounds in rats, and their modulation by glycyl-L-histidyl-L lysine-Cu(II), a potent activator of wound repair. Wound chambers were inserted under the skin of Sprague-Dawley rats and received serial injections of either 2 mg glycyl-L-histidyl-L-lysine-Cu(II) or the same volume of saline. The wound fluid and the neosynthetized connective tissue deposited in the chambers were collected and analyzed for matrix metalloproteinase expression and/or activity. Interstitial collagenase increased progressively in the wound fluid throughout the experiment. Glycyl-L-histidyl-L-lysine-Cu(II) treatment did not alter its activity. Matrix metalloproteinase-9 (gelatinase B) and matrix metalloproteinase 2 (gelatinase A) were the two main gelatinolytic activities expressed during the healing process. Pro-matrix metalloproteinase (pro-form of matrix metalloproteinase)-9 was strongly expressed during the early stages of wound healing (day 3). In the wound fluid, it decreased rapidly and disappeared after day 18, whereas in the wound tissue, matrix metalloproteinase-9 expression persisted in the glycyl-L-histidyl-L-lysine-Cu(II) injected chamber until day 22. Pro-matrix metalloproteinase-2 was expressed at low levels at the beginning of the healing process, increased progressively until day 7, then decreased until day 18. Activated matrix metalloproteinase-2 was present in wound fluid and wound tissue. It increased until day 12, then decreased progressively. Glycyl-L histidyl-L-lysine-Cu(II) injections increased pro-matrix metalloproteinase-2 and activated matrix metalloproteinase-2 during the later stages of healing (days 18 and/or 22). These results demonstrate that various types of matrix metalloproteinases are selectively expressed or activated at the various periods of wound healing. Glycyl-L-histidyl-L-lysine-Cu(II) is able to modulate their expression and might significantly alter wound remodeling. PMID- 10383746 TI - Ultraviolet radiation-induced suppression of natural killer cell activity is enhanced in xeroderma pigmentosum group A (XPA) model mice. AB - Xeroderma pigmentosum group A gene-deficient mice easily develop skin cancers by ultraviolet radiation. Natural killer cells play an important part in tumor surveillance. To study whether ultraviolet radiation-induced suppression of natural killer cell function is involved in the high incidence of skin tumors in patients with xeroderma pigmentosum, we analyzed the number and activity of natural killer cells in ultraviolet B-irradiated xeroderma pigmentosum A model mice. The number of natural killer cells in peripheral blood significantly decreased after ultraviolet B-irradiation only in xeroderma pigmentosum A mice, but those in the spleen were not affected. As compared with the wild-type mice, the xeroderma pigmentosum A mice displayed a higher level of spontaneous splenic natural killer cell activity (10%-15% vs 3%) and inducible natural killer activity (30%-50% vs 20%-25%) after injection of polyinosinic:polycytidylic acid. At 24 h after the last irradiation of three and five daily consecutive exposures to 500 mJ per cm2-ultraviolet B, however, the natural killer activity in xeroderma pigmentosum A mice decreased to 60 and 30% of the preirradiated level, respectively, but it did not in the wild-type mice. The depression of natural killer activity in xeroderma pigmentosum A mice recovered to a normal level at 10 and 15 d after the last irradiation, respectively. The high incidence of skin cancers in xeroderma pigmentosum patients may be mainly due to a defect in the repair of ultraviolet-damaged DNA of cutaneous cells, and possibly also due to an intensified ultraviolet-induced immunosuppression. Moreover, the present study suggests that the enhanced ultraviolet-induced impairment of natural killer function could be partially involved in cancer development. PMID- 10383747 TI - Long-term culture of murine epidermal keratinocytes. AB - The production of transgenic and null mice with skin abnormalities makes it increasingly important to establish cultures of mouse epidermal keratinocytes for in vitro studies. This requires that each cell line be derived from a single mouse and that the cells be carried for multiple passages. Freezing the cells would also be advantageous by allowing comparison of keratinocytes from several mouse lines at the same time. Mouse keratinocytes, however, have been exceedingly difficult to grow as primary cultures, and subculturing these cells has been virtually impossible until now. We describe a gentle dissociation method and a highly supplemented fibroblast conditioned medium that allows us to grow and subculture total mouse keratinocytes for up to 19 subcultures, allowing an increase in cell number of greater than 10 logs. Epidermal keratinocytes from newborn mice were grown on collagen IV coated dishes in murine fibroblast conditioned medium with 0.06 mM calcium and added growth factors. The cells could be passaged, frozen as viable stocks, and induced to differentiate. Morphologically the cultured keratinocytes demonstrated a pattern characteristic of basal cells. Stratified cultures which made mouse keratin 1 and profilaggrin through passage 10 were induced by purging the monolayer cultures of growth factors, then adding medium with 0.15 mM calcium; expression of mouse keratin 1 and profilaggrin was lost by passage 15. The methods explained in detail here should be of great interest to investigators who are now trying to analyze skin phenotypes and expression of markers of epidermal differentiation of their transgenic or knockout mice. PMID- 10383748 TI - Regulation of guanylate-binding protein expression in interferon-gamma-treated human epidermal keratinocytes and squamous cell carcinoma cells. AB - Interferon-gamma is a potent inducer of growth arrest and squamous differentiation of human epidermal keratinocytes in vitro. In order to understand the proximate events regulating interferon-gamma action we studied the relationship between interferon-gamma-mediated induction of a cytoplasmic guanylate-binding protein and the expression of growth and differentiation marker genes in normal and transformed keratinocytes. Induction of guanylate-binding protein mRNA by interferon-gamma was detectable at 4 h, was transcription dependent, and preceded changes in the expression of markers of growth arrest (E2F-1 mRNA downregulation) and differentiation (SQ37 mRNA induction). The Ec50 value for guanylate-binding protein induction (4 units interferon-gamma per ml) was lower than previously reported for SQ37 (40 units interferon-gamma per ml). Guanylate-binding protein mRNA appeared to be only moderately downregulated by modulators of the squamous phenotype such as retinoic acid and transforming growth factor-beta1. In addition, mRNA levels of E2F-1 or SQ37 were not altered in several squamous carcinoma cell lines treated with interferon-gamma. In contrast, guanylate-binding protein mRNA was highly induced in all these cell lines following interferon-gamma treatment. Further analysis of the signal transduction pathway mediating interferon-gamma responses using protein kinase inhibitors indicated that guanylate-binding protein induction in normal human epidermal keratinocyte cells was most likely protein kinase C independent. Our data suggest that more than one postreceptor interferon-gamma signaling pathway exists in keratinocytes and that at least one of these pathways is defective in squamous carcinoma cells. Furthermore, our data demonstrated that the failure of the squamous carcinoma cells to undergo interferon-gamma-induced growth arrest and differentiation is not due to an inherent defect in interferon-gamma receptor activation, but most likely is due to a defect in a non-guanylate-binding protein dependent signaling pathway. PMID- 10383749 TI - Allelic heterogeneity of dominant and recessive COL7A1 mutations underlying epidermolysis bullosa pruriginosa. AB - The inherited mechanobullous disease, dystrophic epidermolysis bullosa, is caused by type VII collagen gene (COL7A1) mutations. We studied six unrelated patients with a distinct clinical subtype of this disease, epidermolysis bullosa pruriginosa, characterized by pruritus, excoriated prurigo nodules, and skin fragility. Mutation analysis using polymerase chain reaction amplification of genomic DNA, heteroduplex analysis and direct nucleotide sequencing demonstrated pathogenetic COL7A1 mutations in each case. Four patients had a glycine substitution mutation on one COL7A1 allele (G1791E, G2242R, G2369S, and G2713R), a fifth was a compound heterozygote for a splice site mutation (5532 + 1G-to-A) and a single base pair deletion (7786delG), and a sixth patient was heterozygous for an out-of-frame deletion mutation (6863del16). This study shows that the molecular pathology in patients with the distinctive clinical features of epidermolysis bullosa pruriginosa is heterogeneous and suggests that other factors, in addition to the inherent COL7A1 mutation(s), may be responsible for an epidermolysis bullosa pruriginosa phenotype. PMID- 10383750 TI - A premature stop codon mutation in the 2B helix termination peptide of keratin 5 in a German epidermolysis bullosa simplex Dowling-Meara case. AB - Epidermolysis bullosa simplex (EBS) is caused by defective assembly of keratin intermediate filaments in basal keratinocytes and recent studies indicated causal mutations in the keratin KRT5 and KRT14 genes. In this study, we describe a novel KRT5 mutation in a German sporadic case of EBS Dowling-Meara. Transition of G to T (nucleotide position 2334) leads to a premature stop codon (E477stop, residue 93 of the 2B helix) in the last residue of the highly conserved helix-termination peptide K/LLEGE of the 2B rod domain of keratin K5. This represents the first premature stop codon mutation identified within the K/LLEGE motif of any disorder reported so far that is caused by keratin mutations. PMID- 10383751 TI - Recurrent COL7A1 mutations in Japanese patients with dystrophic epidermolysis bullosa: positional effects of premature termination codon mutations on clinical severity. Japanese Collaborative Study Group on Epidermolysis Bullosa. PMID- 10383752 TI - Native type I collagen is not a substrate for MMP2 (gelatinase A) PMID- 10383753 TI - Iron acquisition systems in the pathogenic Neisseria. AB - Pathogenic neisseriae have a repertoire of high-affinity iron uptake systems to facilitate acquisition of this essential element in the human host. They possess surface receptor proteins that directly bind the extracellular host iron-binding proteins transferrin and lactoferrin. Alternatively, they have siderophore receptors capable of scavenging iron when exogenous siderophores are present. Released intracellular haem iron present in the form of haemoglobin, haemoglobin haptoglobin or free haem can be used directly as a source of iron for growth through direct binding by specific surface receptors. Although these receptors may vary in complexity and composition, the key protein involved in the transport of iron (as iron, haem or iron-siderophore) across the outer membrane is a TonB dependent receptor with an overall structure presumably similar to that determined recently for Escherichia coli FhuA or FepA. The receptors are potentially ideal vaccine targets in view of their critical role in survival in the host. Preliminary pilot studies indicate that transferrin receptor-based vaccines may be protective in humans. PMID- 10383755 TI - Mechanisms of neisserial serum resistance. AB - Pathogenic Neisseria use a variety of mechanisms to survive the bactericidal action of the complement system. Serum resistance is a crucial virulence factor for the development of severe meningococcal disease, meningococcal meningitis and disseminated gonococcal infection. Furthermore, local inflammation at the site of gonococcal infection exposes the bacteria to moderate concentrations of complement factors. We review current concepts of neisserial serum resistance with emphasis on porins and polysaccharides exposed on the neisserial surface and their interaction with components of normal human serum. PMID- 10383754 TI - Interactions of pathogenic Neisseria with host cells. Is it possible to assemble the puzzle? AB - Neisseria meningitidis and Neisseria gonorrhoeae are human pathogens that have to interact with mucosa and/or cellular barriers for their life cycles to progress. Even though they both give rise to dramatically different diseases, the use of in vitro models has shown that most of the mechanisms mediating cellular interactions are common to N. meningitidis and N. gonorrhoeae. This suggests that bacterial cell interactions may be essential not only for pathogenesis but also for other aspects of the bacterial life cycle that are common to both N. meningitidis and N. gonorrhoeae. This manuscript will review the most recent developments concerning the mechanisms mediating cellular interaction of pathogenic Neisseria and will then try to put them into the perspective of pathogenesis and bacterial life cycle. PMID- 10383756 TI - Genetic redundancy and gene fusion in the genome of the Baker's yeast Saccharomyces cerevisiae: functional characterization of a three-member gene family involved in the thiamine biosynthetic pathway. AB - Redundancy is a salient feature of all living organisms' genome. The yeast genome contains a large number of gene families of previously uncharacterized functions that can be used to explore the functional significance of structural redundancy in a systematic manner. In this work, we describe results on a three-member gene family with moderately divergent sequences (YOL055c, YPL258c and YPR121w ). We demonstrate that two members are isofunctional and encode a hydroxymethylpyrimidine phosphate (HMP-P) kinase (EC 2.7.4.7), an activity required for the final steps of thiamine biosynthesis, whose genes were not previously known in yeast. In addition, we show that the three genes are each composed of two distinct domains, each corresponding to individual genes in prokaryotes, suggesting gene fusion during evolution. The function of the carboxy terminal part of the proteins is not yet understood, but it is not required for HMP-P kinase activity. Expression of all three genes is regulated in the same way. Several other examples of gene fusions exist in the same biosynthetic pathway when eukaryotic genes are compared with prokaryotic ones. PMID- 10383757 TI - Effect of loop deletions on the binding and transport of ferric enterobactin by FepA. AB - The siderophore ferric enterobactin enters Escherichia coli through the outer membrane (OM) porin FepA, which contains an aqueous transmembrane channel that is normally occluded by other parts of the protein. After binding the siderophore at a site within the surface loops, FepA undergoes conformational changes that promote ligand internalization. We assessed the participation of different loops in ligand recognition and uptake by creating and analysing a series of deletions. We genetically engineered 26 mutations that removed 9-75 amino acids from nine loops and two buried regions of the OM protein. The mutations had various effects on the uptake reaction, which we discerned by comparing the substrate concentrations of half-maximal binding (Kd) and uptake (Km): every loop deletion affected siderophore transport kinetics, decreasing or eliminating binding affinity and transport efficiency. We classified the mutations in three groups on the basis of their slight, strong or complete inhibition of the rate of ferric enterobactin transport across the OM. Finally, characterization of the FepA mutants revealed that prior experiments underestimated the affinity of FepA for ferric enterobactin: the interaction between the protein and the ferric siderophore is so avid (Kd < 0.2 nM) that FepA tolerated the large reductions in affinity that some loop deletions caused without loss of uptake functionality. That is, like other porins, many of the loops of FepA are superficially dispensable: ferric enterobactin transport occurred without them, at levels that allowed bacterial growth. PMID- 10383758 TI - Visualization of membrane domains in Escherichia coli. AB - Bacterial membrane and nucleoids were stained concurrently by the lipophilic styryl dye FM 4-64 [N-(3-triethylammoniumpropyl)-4-(6-(4-(diethylamino)phenyl) hexatrienyl)pyridinium dibromide] and 4',6-diamidino-2-phenylindole (DAPI), respectively, and studied using fluorescence microscopy imaging. Observation of plasmolysed cells indicated that FM 4-64 stained the inner membrane preferentially. In live Escherichia coli pbpB cells and filaments, prepared on wet agar slabs, an FM 4-64 staining pattern developed in the form of dark bands. In dividing cells, the bands occurred mainly at the constriction sites and, in filaments, between partitioning nucleoids. The FM 4-64 pattern of dark bands in filaments was abolished after inhibiting protein synthesis with chloramphenicol. It is proposed that the staining patterns reflect putative membrane domains formed by DNA-membrane interactions and have functional implications in cell division. PMID- 10383759 TI - Transcription- and translation-dependent changes in membrane dynamics in bacteria: testing the transertion model for domain formation. AB - Cell cycle events have been proposed to be triggered by the formation of membrane domains in the process of coupled transcription, translation and insertion ('transertion') of nascent membrane and exported proteins. Disruption of domain structure should lead to changes in membrane dynamics. Membrane viscosity of Escherichia coli and Bacillus subtilis decreased after inhibition of protein synthesis by chloramphenicol or puromycin, or of RNA initiation by rifampicin, but not after inhibition of RNA elongation by streptolydigin or amino acid starvation of a stringent strain. The decrease caused by inhibitors of protein synthesis was prevented by streptolydigin if added simultaneously, but was not reversed if added later. The drug-induced decrease in membrane viscosity is energy dependent: it did not happen in KCN-treated cells. All treatments decreasing membrane viscosity also induced nucleoid compaction and fusion. Inhibition of macromolecular synthesis without membrane perturbation caused nucleoids to expand. Changes in membrane dynamics were also displayed during a nutritional shift-down transition that causes imbalance in macromolecular syntheses. The results are consistent with the transertion model, predicting dissipation of membrane domains by termination of protein synthesis or detachment of polysomes from DNA; domain structure is conserved if the transertion process is 'frozen'. PMID- 10383760 TI - Post-transcriptional regulation of the Bacillus subtilis dnaK operon. AB - The heptacistronic dnaK heat shock operon of Bacillus subtilis consists of the genes hrcA, grpE, dnaK, dnaJ, orf35, orf28 and orf50. It is controlled by the CIRCE/HrcA operator/repressor system and specifies three primary transcripts, two of which are processed into three different products. We have analysed the regulatory consequences of this complex transcriptional organization in detail. First, the seven genes were heat induced to different extents at the mRNA level and can be classified into three groups by their induction factors. This differential induction was also reflected at the protein level. Secondly, the cellular amounts of the proteins HrcA, DnaK and DnaJ in B. subtilis differed drastically both under non-heat shock conditions and after thermal upshock. Thirdly, Northern blot analyses demonstrated that an mRNA-processing reaction generating products of differential stabilities plays an essential role during the regulation of gene expression. A crucial factor determining the low stability of two transcripts is the presence of the CIRCE element at their 5' ends. We demonstrate that CIRCE leads to the destabilization of mRNAs, but only if it is located in the immediate vicinity of a Shine-Dalgarno sequence. These results show that B. subtilis is using various, especially post-transcriptional, regulatory mechanisms to fine tune the expression of the individual genes of the heptacistronic dnaK operon. PMID- 10383761 TI - The response to stationary-phase stress conditions in Escherichia coli: role and regulation of the glutamic acid decarboxylase system. AB - Inducible bacterial amino acid decarboxylases are expressed at the end of active cell division to counteract acidification of the extracellular environment during fermentative growth. It has been proposed that acid resistance in some enteric bacteria strictly relies on a glutamic acid-dependent system. The Escherichia coli chromosome contains distinct genes encoding two biochemically identical isoforms of glutamic acid decarboxylase, GadA and GadB. The gadC gene, located downstream of gadB, has been proposed to encode a putative antiporter implicated in the export of gamma-aminobutyrate, the glutamic acid decarboxylation product. In the present work, we provide in vivo evidence that gadC is co-transcribed with gadB and that the functional glutamic acid-dependent system requires the activities of both GadA/B and GadC. We also found that expression of gad genes is positively regulated by acidic shock, salt stress and stationary growth phase. Mutations in hns, the gene for the histone-like protein H-NS, cause derepressed expression of the gad genes, whereas the rpoS mutation abrogates gad transcription even in the hns background. According to our results, the master regulators H-NS and RpoS are hierarchically involved in the transcriptional control of gad expression: H-NS prevents gad expression during the exponential growth whereas the alternative sigma factor RpoS relieves H-NS repression during the stationary phase, directly or indirectly accounting for transcription of gad genes. PMID- 10383762 TI - The haemolysin-secreting ShlB protein of the outer membrane of Serratia marcescens: determination of surface-exposed residues and formation of ion permeable pores by ShlB mutants in artificial lipid bilayer membranes. AB - The ShlB protein in the outer membrane of Serratia marcescens is the only protein known to be involved in secretion of the ShlA protein across the outer membrane. At the same time, ShlB converts ShlA into a haemolytic and a cytolytic toxin. Surface-exposed residues of ShlB were determined by reaction of an M2 monoclonal antibody with the M2 epitope DYKDDDDK inserted at 25 sites along the entire ShlB polypeptide. The antibody bound to the M2 epitope at 17 sites in intact cells, which indicated surface exposure of the epitope, and to 23 sites in isolated outer membranes. Two insertion mutants contained no ShlB(M2) protein in the outer membrane. The ShlB derivatives activated and/or secreted ShlA. To gain insights into the secretion mechanism, we studied whether highly purified ShlB and ShlB deletion derivatives formed pores in artificial lipid bilayer membranes. Wild type ShlB formed channels with very low single channel conductance that rarely assumed an open channel configuration. In contrast, open channels with a considerably higher single channel conductance were observed with the deletion mutants ShlB(Delta65-186), ShlB(Delta87-153), and ShlB(Delta126-200). ShlB(Delta126-200) frequently formed permanently open channels, whereas the conductance caused by ShlB(Delta65-186) and ShlB(Delta87-153) did not assume a stationary value, but fluctuated rapidly between open and closed configurations. The results demonstrate the orientation of large portions of ShlB in the outer membrane and suggest that ShlB may function as a specialized pore through which ShlA is secreted. PMID- 10383763 TI - Molecular analysis of the cytolytic protein ClyA (SheA) from Escherichia coli. AB - Escherichia coli K-12 carries a gene for a protein denoted ClyA or SheA that can mediate a cytolytic phenotype. The ClyA protein is not expressed at detectable levels in most strains of E. coli, but overproduction suitable for purification was accomplished by cloning the structural gene in an hns mutant strain. Highly purified ClyA protein was cytotoxic to macrophage cells in culture and caused detachment and lysis of the mammalian cells. Results from osmotic protection assays were consistent with the suggestion that the protein formed pores with a diameter of up to 3 nm. Using Acholeplasma laidlawii cells and multilamellar liposomes, we studied the effect of ClyA on membranes with varying compositions and in the presence of different ions. ClyA induced cytolytic release of the fluorescent tracer from carboxyfluorescein-loaded liposomes, and the release was stimulated if cholesterol was present in the membranes whereas addition of calcium had no effect. Pretreatment of the ClyA protein with cholesterol inhibited the pore formation, suggesting that ClyA could bind to cholesterol. Efficient coprecipitation of ClyA with either cholesterol or 1,2,3 trioctadecanoylglycerol in aqueous solutions showed that ClyA directly interacted with the hydrophobic molecular aggregates. We tested the possible functional importance of selected ClyA protein regions by site-directed mutagenesis. Defined mutants of ClyA were obtained with alterations in postulated transmembrane structures in the central part and in a postulated membrane-targeting domain in the C-terminal part. Our results were consistent with the suggestion that particular amphiphilic segments are required for ClyA activity. We propose that these domains are necessary for ClyA to form pores. PMID- 10383765 TI - Utilization of exogenous folate in the human malaria parasite Plasmodium falciparum and its critical role in antifolate drug synergy. AB - The antifolate combination pyrimethamine/sulphadoxine (PYR/SDX; Fansidar) is frequently used to combat chloroquine-resistant malaria. Its success depends upon pronounced synergy between the two components, which target dihydrofolate reductase (DHFR) and dihydropteroate synthetase (DHPS) in the folate pathway. This synergy permits clearance of parasites resistant to either drug alone, but its molecular basis is still unexplained. Plasmodium falciparum can use exogenous folate, which is normally present in vivo, bypassing SDX inhibition of DHPS and, apparently, precluding synergy under these conditions. However, we have measured parasite inhibition by SDX/PYR combinations in assays in which folate levels are strictly controlled. In parasites that use exogenous folate efficiently, SDX inhibition can be restored by levels of PYR significantly lower than those required to inhibit DHFR. Isobolograms show that the degree of synergy between PYR and SDX is highly dependent upon prevailing folate concentrations and are indicative of PYR acting to block folate uptake and/or utilization. No significant synergy was observed at physiological drug levels when PYR/SDX acted on purified DHFR, whether wild type or mutant. We conclude that the primary basis for antifolate synergy in these organisms arises from PYR targeting a site (or sites) in addition to DHFR, which restores DHPS as a relevant target for SDX. PMID- 10383764 TI - Dimerization of the Agrobacterium tumefaciens VirB4 ATPase and the effect of ATP binding cassette mutations on the assembly and function of the T-DNA transporter. AB - The Agrobacterium tumefaciens VirB4 ATPase functions with other VirB proteins to export T-DNA to susceptible plant cells and other DNA substrates to a variety of prokaryotic and eukaryotic cells. Previous studies have demonstrated that VirB4 mutants with defects in the Walker A nucleotide-binding motif are non-functional and exert a dominant negative phenotype when synthesized in wild-type cells. This study characterized the oligomeric structure of VirB4 and examined the effects of Walker A sequence mutations on complex formation and transporter activity. VirB4 directed dimer formation when fused to the amino-terminal portion of cI repressor protein, as shown by immunity of Escherichia coli cells to lambda phage infection. VirB4 also dimerized in Agrobacterium tumefaciens, as demonstrated by the recovery of a detergent-resistant complex of native protein and a functional, histidine-tagged derivative by precipitation with anti-His6 antibodies and by Co2+ affinity chromatography. Walker A sequence mutants directed repressor dimerization in E. coli and interacted with His-VirB4 in A. tumefaciens, indicating that ATP binding is not required for self-association. A dimerization domain was localized to a proposed N-terminal membrane-spanning region of VirB4, as shown by the dominance of an allele coding for the N-terminal 312 residues and phage immunity of host cells expressing cI repressor fusions to alleles for the first 237 or 312 residues. A recent study reported that the synthesis of a subset of VirB proteins, including VirB4, in agrobacterial recipients has a pronounced stimulatory effect on the virB-dependent conjugal transfer of plasmid RSF1010 by agrobacterial donors. VirB4'312 suppressed the stimulatory effect of VirB proteins for DNA uptake when synthesized in recipient cells. In striking contrast, Walker A sequence mutants contributed to the stimulatory effect of VirB proteins to the same extent as native VirB4. These findings indicate that the oligomeric structure of VirB4, but not its capacity to bind ATP, is important for the assembly of VirB proteins as a DNA uptake system. The results of these studies support a model in which VirB4 dimers or homomultimers contribute structural information for the assembly of a transenvelope channel competent for bidirectional DNA transfer, whereas an ATP-dependent activity is required for configuring this channel as a dedicated export machine. PMID- 10383767 TI - Intracellular growth and metacyclogenesis defects in Trypanosoma cruzi carrying a targeted deletion of a Tc52 protein-encoding allele. AB - We have identified previously a Trypanosoma cruzi gene encoding a protein named Tc52 sharing structural and functional properties with the thioredoxin and glutaredoxin protein family involved in thiol-disulphide redox reactions. Furthermore, we have reported that Tc52 also played a role in T. cruzi-associated immunosuppression observed during Chagas' disease. In an effort to understand further the biological role of Tc52, we used a gene-targeted deletion strategy to create T. cruzi mutants. Although T. cruzi tolerates deletion of one wild-type Tc52 allele, deletion of both genes is a lethal event, indicating that at least one active Tc52 gene is required for parasite survival. Monoallelic disruption of Tc52 (Tc52+/-) resulted in the production of T. cruzi lines that express less Tc52 mRNA and produced lower amounts of Tc52 protein compared with wild-type cells. In axenic cultures, growth rates of epimastigote forms bearing an interrupted allele were not different from those of wild-type parasites. Furthermore, monoallelic disruption of the Tc52 gene did not modify the growth rate of epimastigotes or their sensitivity to inhibition by benznidazole and nifurtimox, the two drugs used to treat Chagasic patients. Moreover, the antimonial drug SbIII, which is known, at least in Leishmania parasites, to be conjugated to a thiol and extruded by an ATP-coupled pump, had a similar effect on wild-type and mutant parasites, being equally sensitive. Hence, parasite drug sensitivity was also observed in clones overexpressing the Tc52 protein as well as in those carrying an antisense plasmid construct. Surprisingly, a significant impairment of the ability of epimastigotes carrying a Tc52 single gene replacement or antisense construct to differentiate into metacyclic trypomastigotes and to proliferate in vitro and in vivo was observed, whereas no significant enhancement of these biological properties was seen in the case of parasites that overexpress Tc52 protein. Moreover, functional complementation of Tc52+/- single mutant or selection of antisense revertant clones demonstrated that the phenotype observed is a direct consequence of Tc52 gene manipulation. Taken together, these results may suggest that Tc52 could participate among other factors in the phenotypic expression of T. cruzi virulence. PMID- 10383766 TI - Stress factors acting at the level of the plasma membrane induce transcription via the stress response element (STRE) of the yeast Saccharomyces cerevisiae. AB - A variety of stress factors induces transcription via the stress response element (STRE) present in control regions of a number of genes of the yeast Saccharomyces cerevisiae. Induction of transcription involves nuclear translocation of the STRE binding transcription activators Msn2p and Msn4p. The primary cellular events triggering this translocation are presently not well understood. In this investigation, we have observed that a number of factors acting at the level of the yeast plasma membrane, including the antifungal agent nystatin, the steroidal alkaloid tomatine, benzyl alcohol, a number of detergents and the plasma membrane H+-ATPase inhibitor diethylstilbestrol or mutations in the PMA1 gene encoding the plasma membrane ATPase, induce Msn2p nuclear accumulation and STRE-dependent transcription. At least some of the stress factors acting via STREs cause an increase in plasma membrane permeability, leading to a decrease in membrane potential, which might be a primary cellular stress signal. A decrease in internal pH triggered by permeabilization of the plasma membrane or a change in cAMP levels are at least not obligatory factors in intracellular stress signal transduction. The signal transduction pathway transmitting the signal generated at the plasma membrane to Msn2p is still unknown. PMID- 10383768 TI - Introduction of the exopolysaccharide gene cluster from Streptococcus thermophilus Sfi6 into Lactococcus lactis MG1363: production and characterization of an altered polysaccharide. AB - Streptococcus thermophilus Sfi6 produces an exopolysaccharide (EPS) composed of glucose, galactose and N-acetylgalactosamine in the molar ratio of 1:2:1. The genes responsible for the EPS biosynthesis have been isolated previously and found to be clustered in a 14.5 kb region encoding 13 genes. Transfer of this gene cluster into a non-EPS-producing heterologous host, Lactococcus lactis MG1363, yielded an EPS with a similar high molecular weight, but a different structure from the EPS from the native host. The structure of the recombinant EPS was determined by means of 1H homonuclear and 1H-13C heteronuclear two dimensional nuclear magnetic resonance (NMR) spectra and was found to be --> 3) beta-D-Glcp-(1 --> 3)-alpha-D-Galp-(1 --> 3)-beta-D-Galp-(1 --> as opposed to --> 3)[alpha-D-Galp-(1 --> 6)]-beta-D-Glcp-(1 --> 3)-alpha-D-GalpNAc-(1 --> 3)-beta-D Galp-(1 --> for the wild-type S. thermophilus Sfi6. Furthermore, L. lactis MG1363 (pFS101) was also lacking a UDP-N-acetylglucosamine C4-epimerase activity, which would provide UDP-GalNAc for a GalNAc incorporation into the EPS and probably caused the substitution of GalNAc by Gal in the recombinant EPS. This modification implies that (i) bacterial glycosyltransferases could potentially have multiple specificities for the donor and the acceptor sugar molecule; and (ii) the repeating unit polymerase can recognize and polymerize a repeating unit that differs in the backbone as well as in the side-chain from its native substrate. PMID- 10383769 TI - Antiterminator-dependent modulation of transcription elongation rates by NusB and NusG. AB - Ribosomal RNA is transcribed about twice as fast as messenger RNA in vivo, and this increased transcription rate requires the rrn boxA antitermination system. Because several Nus factors have been implicated in rrn antitermination, we have examined the role of NusB, NusE and NusG in controlling the rate of rrn boxA mediated transcript elongation. In vivo RNA polymerase transcription rates were determined by measuring the rate of appearance of lacZ transcript using a plasmid that contained an inducible T7 promoter fused to the rrn boxA sequence followed by the lacZ gene. This plasmid was introduced into Escherichia coli mutant strains that can be conditionally depleted of NusG, or that carry a deficient nusB gene or a nusE mutation. We found that, in addition to the rrn boxA antiterminator sequence, both NusG and NusB were required to maintain the high transcription rate. The nusE mutation used in this study may be specific for lambda antitermination, as it did not influence the boxA-mediated increase in transcription rate. PMID- 10383770 TI - The involvement of NAD(P)H dehydrogenase subunits, NdhD3 and NdhF3, in high affinity CO2 uptake in Synechococcus sp. PCC7002 gives evidence for multiple NDH 1 complexes with specific roles in cyanobacteria. AB - Random gene tagging was used to obtain new mutants of the marine cyanobacterium, Synechococcus sp. PCC7002, with defects in the CO2-concentrating mechanism (CCM). Two of these mutants, K22 and A41, showed poor growth at limiting CO2. Isolation and sequencing of a 6. 6 kb genomic region revealed the existence of five potential protein-coding regions, all arranged in the same transcriptional direction. These regions code for an RbcR homologue, NdhF3 (subunit 5 of type 1 NAD(P)H dehydrogenase; NDH-1 complex), NdhD3 (subunit 4 of NDH-1), ORF427 and ORF133 (hypothetical proteins). Insertional mutants in ndhD3, ndhF3 and ORF427, like A41 and K22, were all incapable of inducing high-affinity CO2 uptake and were not fully capable of inducing high-affinity HCO3- transport. ndhD3 and ndhF3 mutants displayed P700 re-reduction rates identical to wild-type cells, suggesting that NdhD3 is part of a specific NDH-1 complex that is not involved in photosynthetic cyclic electron transport. Thus, it is feasible that NdhD3, NdhF3 and ORF427 might form part of a novel NDH-1 complex located on the cytoplasmic membrane and involved in tightly coupled energization of high-affinity CO2 transport. The possibility of multiple, functionally distinct NDH-1 complexes in cyanobacteria is discussed. PMID- 10383771 TI - Type IV pili of pathogenic Neisseriae elicit cortical plaque formation in epithelial cells. AB - The pathogenic Neisseriae Neisseria meningitidis and Neisseria gonorrhoeae, initiate colonization by attaching to host cells using type IV pili. Subsequent adhesive interactions are mediated through the binding of other bacterial adhesins, in particular the Opa family of outer membrane proteins. Here, we have shown that pilus-mediated adhesion to host cells by either meningococci or gonococci triggers the rapid, localized formation of dramatic cortical plaques in host epithelial cells. Cortical plaques are enriched in both components of the cortical cytoskeleton and a subset of integral membrane proteins. These include: CD44v3, a heparan sulphate proteoglycan that may serve as an Opa receptor; EGFR, a receptor tyrosine kinase; CD44 and ICAM-1, adhesion molecules known to mediate inflammatory responses; f-actin; and ezrin, a component that tethers membrane components to the actin cytoskeleton. Genetic analyses reveal that cortical plaque formation is highly adhesin specific. Both pilE and pilC null mutants fail to induce cortical plaques, indicating that neisserial type IV pili are required for cortical plaque induction. Mutations in pilT, a gene required for pilus mediated twitching motility, confer a partial defect in cortical plaque formation. In contrast to type IV pili, many other neisserial surface structures are not involved in cortical plaque induction, including Opa, Opc, glycolipid GgO4-binding adhesins, polysialic acid capsule or a particular lipooligosaccharide variant. Furthermore, it is shown that type IV pili allow gonococci to overcome the inhibitory effect of heparin, a soluble receptor analogue, on gonococcal invasion of Chang and A431 epithelial cells. These and other observations strongly suggest that type IV pili play an active role in initiating neisserial infection of the mucosal surface in vivo. The functions of type IV pili and other neisserial adhesins are discussed in the specific context of the mucosal microenvironment, and a multistep model for neisserial colonization of mucosal epithelia is proposed. PMID- 10383772 TI - Tinea capitis: an overview with emphasis on management. AB - Tinea capitis is perhaps the most common mycotic infection in children. In North America the epidemiology of tinea capitis has changed so that Trichophyton tonsurans now predominates over Micro-sporum audouinii. With this transition the utility of the Wood's light for diagnosis has been reduced since T. tonsurans infection is Wood's light negative. Griseofulvin has been the mainstay of therapy for the last 40 years. The newer antifungal agents-itraconazole, terbinafine, and fluconazole-appear to be effective and safe for the treatment of tinea capitis. When tinea capitis is due to T. tonsurans or other endothrix species the following regimens have been used: itraconazole continuous regimen (5 mg/kg/day for 4 weeks), itraconazole pulse regimen with capsules (5 mg/kg/day for 1 week plus 1-3 pulses 3 weeks apart), and itraconazole pulse regimen with oral solution (3 mg/kg/day for 1 week plus 1-3 pulses 3 weeks apart). With terbinafine tablets the continuous regimen (>40 kg body weight, 250 mg/day; 20-40 kg, 125 mg/day; and <20 kg, 125 mg/day) is given for 2 to 4 weeks. Fluconazole tablets or oral suspension (6 mg/kg/day) were administered for 20 days in one trial. Another possibility may be 6 mg/kg/day for 2 weeks and evaluating the scalp 4 weeks later. An extra week of therapy (6 mg/kg/day) can be administered if clinically indicated at that time. A once-weekly regimen may also be effective. When ectothrix organisms (e.g., Microsporum canis) are present, a longer duration of therapy may be required. The data suggest that the newer agents are effective, safe with few adverse effects, and have a high benefit:risk ratio. It remains to be seen to what extent griseofulvin will be superseded for the treatment of tinea capitis. Adjunctive therapies may help decrease the risk of infection to other individuals. Appropriate measures should be taken to reduce the possibility of reinfection. PMID- 10383773 TI - Stress and family satisfaction in parents of children with facial port-wine stains. AB - A cross-sectional survey was employed to assess parenting stress, family satisfaction, and parental concerns and to determine predictors of stress in parents of children with port-wine stains (PWSs). The participants were 46 parents of 24 children receiving treatment with pulsed dye laser photocoagulation for facial PWS at an outpatient dermatology clinic based at a university medical center. Outcome measures used were self-report instruments assessing psychosocial adjustment (Parenting Stress Index, Family Satisfaction Scale, and Parental Concerns Questionnaire). As a group, parents scored in the average range on the stress and family satisfaction measures when compared with a normative sample; five parents (11%) scored in the clinical range for stress. Forty-nine percent of the variance in parenting stress was accounted for by four variables: the child's age (beta = 0.34; p = 0.031), the parents' degree of family satisfaction (beta = 0.27; p = 0.077), the level of parental concern regarding the child's facial PWS (beta = 0.45; p = 0.005), and the parents' satisfaction with staff communication (beta = -0. 51; p = 0.002). The data suggest that while, as a group, parents of children with a facial PWS report to be in the average range for psychological stress, some do not fare as well as others. Factors associated with lower stress include younger children, more family cohesion and adaptation, fewer parental concerns, and greater satisfaction with parent-staff communication. The potential for the development of medical complications and psychological problems over time suggests the need for treatment of the PWS at an early age. Health care providers should be prepared to screen for clinical levels of distress and to refer parents for psychological intervention when needed. PMID- 10383774 TI - The New Moms Project: educating mothers about sun protection in newborn nurseries. AB - Sun protection habits should begin early in life and be taught as part of routine preventive health care. Early teaching of parents aims to introduce an easily achieved means of sun protection with the goal of instilling these practices as habits in the parents and their young children. We developed a maternity nurse led intervention for 187 mothers at newborn nurseries in Falmouth, Massachusetts, combining educational material and personal discussions. One year after the intervention we successfully contacted 73% of the mothers. Nearly 90% recalled the informational program and equal numbers stated that receiving educational materials in the newborn nursery was timely. Nearly two-thirds of mothers reported that this was the only sun protection information received from a provider in the past year. PMID- 10383775 TI - Prevalence and new phenotypic and radiologic findings in congenital onychodysplasia of the index finger. AB - Congenital onychodysplasia of the index finger (COIF) is a rare condition characterized by dysplastic changes in the nail with variable phenotypic manifestations. Its prevalence is unknown. We describe three newborn patients with various clinical and radiologic expressions of this entity. The first, born to a mother treated with multiple antidepressant drugs, had bilateral nail dysplasia of the index finger without bony involvement. The second presented with bilateral hypoplastic nail of the index finger and bilateral symphalangism of the fifth finger; aplastic distal phalanges of the second finger was also noted radiologically. The third baby (born to a diabetic mother treated with insulin), had micronychia and brachydactyly of the right index finger and syndactyly of the second and third fingers; the radiologic finding was bilateral brachymesophalangia of the index finger. It would seem that, besides the nail dysplasia of the index finger, COIF may also be associated with bone dysplasia of the same finger. PMID- 10383776 TI - Childhood immunobullous disease following a second organ transplant. AB - Immunobullous diseases are quite unusual in children. We report two children who developed immunobullous disease shortly after a second solid organ transplantation. One child had IgA pemphigus vegetans, the other bullous pemphigoid. We hypothesize that the repeat organ transplantation served as a "booster" immunization for autoantibody production. Both children developed their immunobullous condition while being treated with immunosuppressive drugs that are used to treat immunobullous disorders. PMID- 10383777 TI - Primary disseminated varicella presenting as an acute abdomen. AB - We report a patient admitted with acute abdominal pain initially thought to be due to pancreatitis of unclear etiology. Later during his hospitalization he was diagnosed with primary varicella infection. The association between varicella and systemic multiorgan disease needs to be recognized in both immunocompetent and immunocompromised patients. A prompt diagnosis prevents delay in the treatment of varicella, as well as in monitoring for and preventing complications of disseminated infection. PMID- 10383778 TI - The spectrum of epidermal nevi: a case of verrucous epidermal nevus contiguous with nevus sebaceus. AB - During the normal development of skin, pluripotential cells give rise to keratinocytes, sebaceous glands, hair follicles, apocrine glands, and eccrine glands. In epidermal nevi, these components emerge in an abnormal mixture within a circumscribed site. Many authors have categorized epidermal nevi based on their predominant component; however, there is often notable overlap that occurs within a single area or within contiguous areas. We report a verrucous epidermal nevus contiguous to a nevus sebaceus of Jadassohn. The categories of epidermal nevi are somewhat artificial. Our case supports the view that epidermal nevi have a spectrum of manifestations, including verrucous epidermal nevi and nevus sebaceus of Jadassohn. PMID- 10383779 TI - Neutrophilic dermatosis-associated sterile chronic multifocal osteomyelitis in pediatric patients: case report and review. AB - Atypical pyoderma gangrenosum (PG) and Sweet syndrome are neutrophilic dermatoses that share some common features. Sterile chronic recurrent multifocal osteomyelitis is a rare association of these neutrophilic dermatoses that has only been reported in children. We report a 3-year-old girl who initially presented with pain in her left hand and right leg. Roentgenograms and bone scan revealed findings of multifocal osteomyelitis affecting both femurs, the right tibia, left clavicle, right eighth costochondral junction, and left ulna. She was treated with antibiotics without improvement. Bone biopsy of the left ulna revealed histologic changes consistent with osteomyelitis, however, all cultures for bacteria, mycobacteria, and fungi were negative. She subsequently developed an ulcer surrounded by a violaceous, undermined border at the site of the bone biopsy, which also did not improve during antibiotic treatment. A biopsy specimen from this lesion demonstrated a dense perivascular and periappendageal infiltrate of neutrophils within the dermis and edema of the papillary dermis compatible with a neutrophilic dermatosis. She was treated with oral prednisone which resulted in resolution of skin lesions, bone pain, and soft tissue swelling. This case further documents the association between PG or Sweet syndrome and multifocal sterile osteomyelitis. PMID- 10383780 TI - Radius hypoplasia, radial palsy, and aplasia cutis due to amniotic band syndrome. AB - Amniotic band syndrome is one of the many causes of aplasia cutis congenita. It is usually seen as a constriction band surrounding a limb or as a membrane that adheres to some part of the body. This syndrome can be associated with various malformations. An infant with amniotic adhesions producing aplasia cutis, radial palsy, and hypoplasia of the radius is presented. Early treatment led to total functional recovery of the affected limb. PMID- 10383782 TI - Blue rubber bleb nevus (Bean syndrome): evolution of four cases and clinical response to pharmacologic agents. AB - Blue rubber bleb nevus (BRBN) syndrome is a rare disorder characterized by distinctive cutaneous and gastrointestinal vascular malformations. The latter may lead to bleeding complications. We followed four affected children for at least 5 years. The evolution of their disease and the value of pharmacologic agents (steroids, interferon) in the management of some of these patients are discussed. PMID- 10383781 TI - Pili torti with congenital deafness (Bjornstad syndrome): a case report. AB - We report Bjornstad syndrome in a 5-year-old girl with severe bilateral congenital loss of hearing and pili torti. The mode of inheritance of this rare syndrome seems to be heterogeneous. A maternal uncle of the patient was deaf from birth and his hair had shown the same abnormalities at the same age; an autosomal recessive transmission can be assumed. PMID- 10383783 TI - Treatment of molluscum contagiosum with potassium hydroxide: a clinical approach in 35 children. AB - Potassium hydroxide (KOH) is a strong alkali that has long been known to digest proteins, lipids, and most other epithelial debris of skin scrapings to identify fungal infections. To our knowledge, KOH has never been used for the treatment of molluscum contagiosum (MC). We evaluated 35 children with MC for the clinical effectiveness of treatment with topical 10% KOH aqueous solution. The solution was applied by the parents of affected children, twice daily, on each MC lesion. The therapy was continued until all lesions underwent inflammation and superficial ulceration. Thirty-two of 35 patients achieved complete clinical cure after a mean treatment period of 30 days. Three children discontinued treatment: two reported severe stinging of the lesions and refused further applications; the other, with giant MC lesions, developed a secondary infection with prolonged treatment. Therapy with KOH was found to be effective and safe in the treatment of MC in children. PMID- 10383784 TI - Cranial fasciitis of childhood. PMID- 10383785 TI - What syndrome is this? Macrocephaly with cutis marmorata, hemangioma, and syndactyly syndrome. PMID- 10383786 TI - Vascular tumors and vascular malformations: are we at the dawn of a better knowledge? PMID- 10383787 TI - Cutis verticis gyrata in a child with Turner syndrome. PMID- 10383788 TI - Tazarotene gel in childhood Darier disease. PMID- 10383789 TI - Bleeding from beneath the nails: an unusual artifactual disease in a child. PMID- 10383790 TI - Pansclerotic morphea of childhood-follow-up over 6 years. PMID- 10383791 TI - A portrait in history. Sir Charles Bell. Artist extraordinaire. PMID- 10383792 TI - College of American Pathologists Conference XXXIII on transfusion medicine performance improvement. Introduction. PMID- 10383793 TI - Requirements for accreditation by the College of American Pathologists Laboratory Accreditation Program. AB - The College of American Pathologists Laboratory Accreditation Program expects a participant laboratory or laboratory section to be able to demonstrate that it is in compliance with the Standards for Laboratory Accreditation. The program expects laboratories to demonstrate that they are continually taking steps to identify and correct deficient areas and improve performance, in compliance with the Clinical Laboratory Improvement Amendments of 1988 regulatory requirements, particularly those pertaining to proficiency testing performance, and participating as inspectors in the accreditation process. PMID- 10383794 TI - What do the accreditation organizations expect? American Association of Blood Banks. AB - In 1958, the American Association of Blood Banks introduced the first edition of Standards for Blood Banks and Transfusion Services. That same year, the association implemented the Inspection and Accreditation Program. This program served the association well for 40 years; however, factors such as the application of Current Good Manufacturing Practices by the Food and Drug Administration, the implementation of the Clinical Laboratory Improvement Amendments of 1988 by the Health Care Financing Administration, managed care, competition, and increased cost pressures have changed the way the blood banking community conducts its business. In the early 1990s the board of directors recognized the need to reevaluate the Inspection and Accreditation Program and developed a strategic plan for implementation of a new accreditation program, with an emphasis on prevention rather than detection of errors. The first step in the process was the development of the Accreditation Program Committee. The committee was charged to develop and coordinate a program that would bring the accreditation process in tune with the current climate of blood banking and move it into the 21st century. The board charged the committee with the development of a program that recognizes the differences and similarities within the diverse groups of American Association of Blood Banks institutional members and to take into consideration how they do business and respond to regulations, standards, and other requirements. PMID- 10383795 TI - Joint Commission on Accreditation of Healthcare Organizations' expectations for transfusion medicine in health care organizations. AB - This article provides an overview of the Joint Commission on Accreditation of Healthcare Organizations' standards related to transfusion medicine found in its hospital, laboratory, and home care accreditation manuals. Hospital standards focus on the review and evaluation of the entire transfusion process from the order through the outcome to the patient, with special attention to the blood use review process. Laboratory standards provide the structure for the detailed review of the technical procedures and practices for collecting, processing, storing, testing, and transporting blood products. Home care standards relate to policies and procedures, infection control practices, education of the patient and family, and monitoring of adverse events and complications for transfusions of blood products performed in the home. PMID- 10383796 TI - Blood safety. The Food and Drug Administration's role. AB - The federal government has a role in protecting consumers from threats to the blood supply. These threats range from the residual risks of known infection, to emerging infections that may contaminate the blood supply, to complacency in the area of adherence to product standards and Current Good Manufacturing Practices. The Food and Drug Administration is tasked with oversight of the safety of the nation's blood supply. The Food and Drug Administration's regulation of blood and blood components includes preapproval for products entering interstate commerce and adherence to Current Good Manufacturing Practices for all blood components. Oversight for Current Good Manufacturing Practices compliance includes inspection and reporting requirements. These regulatory programs are coupled with the Public Health Service's ongoing research and epidemiological surveillance. The goals of these programs are to prevent another infectious disease epidemic from infecting the blood supply as did the acquired immunodeficiency syndrome in the early 1980s and to further improve blood safety. PMID- 10383797 TI - Health Care Financing Administration/clinical laboratory improvement amendments of 1988. AB - The Health Care Financing Administration has introduced new concepts for the Clinical Laboratory Improvement Amendments of 1988 survey program. Surveyors now look at the laboratory as a whole as opposed to each regulatory requirement independently. This quality assurance approach allows the surveyor to assess the laboratory's ability to provide quality test results as well as identify and correct its own problems. Significant problems in laboratories are more easily identified using this method. Another new concept allows good performing laboratories to go longer between on-site surveys by completing a self-assessment questionnaire on alternate cycles. The Health Care Financing Administration is asking both surveyors and laboratories to evaluate these new approaches. The Health Care Financing Administration continues to work with the Food and Drug Administration through our Memorandum of Understanding to assess Food and Drug Administration requirements in hospitals and laboratories that provide transfusion services. Transfusion-related fatalities must be reported to the Food and Drug Administration and may be investigated by either the Health Care Financing Administration or the Food and Drug Administration. For fatalities needing investigation by both agencies, every effort will be made to conduct these jointly. PMID- 10383798 TI - An overview of state efforts to improve transfusion medicine. The New York state model. AB - OBJECTIVE: New York State must provide effective oversight for more than 400 facilities that provide blood services, ensuring the safety of transfusion recipients. PROGRAM STRUCTURE: Oversight is based on educated, trained program staff; a council of transfusion medicine experts; regulations governing laboratories and blood banking services; proficiency testing; requirements for reporting transfusion-related errors and incidents; tracking collection, testing, and disposition of all blood; inspections; partnership with the Food and Drug Administration; and investigation of incident reports. Policy letters and guidelines (eg, umbilical cord blood, transfusion-associated infections, intraoperative blood recovery) supplement regulations. OBSERVATIONS: Standards are maintained through education by a responsive, proactive oversight program. Areas warranting stricter regulation or supplemental educational guidelines are identified by periodic assessment of standards of practice and review of reported information. CONCLUSION: The multifaceted oversight program contributes to efforts to improve transfusion medicine. PMID- 10383799 TI - Transfusion medicine's role in hospital performance improvement. An administrator's view. AB - Historically, hospital administrators have viewed all laboratory sections as cost centers. The major expenses in the transfusion service are those associated with labor and blood products. However, few administrators take the time to look past this cost to see the impact of an active transfusion medicine section in other areas of the hospital. This article examines the impact of inventory management, blood component utilization and waste, group purchasing, and new program implementation on transfusion service expense and revenue. PMID- 10383800 TI - How to improve transfusion medicine. A treating physician's perspective. AB - As transfusion medicine becomes more complex, cooperative strategies are gaining increasing importance in relaying information to the treating physician and in incorporating the treating physician into the education and quality control processes. The broad domain of transfusion medicine is illustrated by the variety of disciplines involved in defining the use of products such as fresh frozen plasma and the newly released solvent-detergent-treated plasma, fibrin glue and highly purified fibrin sealant, and leukoreduced and irradiated blood products. Cooperative efforts among physicians and other personnel of multiple disciplines are essential to ensure appropriate use and continuous evaluation of blood products. PMID- 10383801 TI - Performance improvement in transfusion medicine. What do nurses need and want? AB - Transfusion medicine is a complex process dependent on a variety of professionals interacting effectively and efficiently across time and distance. To perform safely, professionals depend on their own knowledge and skills, the knowledge and skills of others, and the overall effectiveness of operating systems. Nursing is an essential link in the process. To be effective, nurses need to practice in environments that recognize the importance of reducing error and improving safety through use of nonpunitive system approaches to analyzing near misses and errors. The "off-with-their-heads" approach must be eliminated. To increase efficiency, pathologists and nurses should collaborate on form development, evaluation, and implementation. Documentation regarding transfusions needs to be simplified and coordinated. Knowledgeable staff is an essential element of safe systems. Basic knowledge should never be assumed. Mechanisms to monitor knowledge of key processes along with ongoing feedback and remediation are necessary to maximize performance. Working together, nursing and transfusion specialists will improve transfusion services. PMID- 10383802 TI - Immunohistochemical profile of myogenin and MyoD1 does not support skeletal muscle lineage in alveolar soft part sarcoma. AB - BACKGROUND: The histogenesis of alveolar soft part sarcoma remains elusive. Myogenic origin is favored, although conflicting data on immunohistochemical demonstration of muscle-associated markers exist. Myogenin and MyoD1, transcription factors of the myogenic determination family, have crucial roles in commitment and differentiation of mesenchymal progenitor cells to myogenic lineage and in maintenance of skeletal muscle phenotype. Their immunohistochemical detection is specific in characterization of rhabdomyosarcoma. METHODS: Antibodies for myogenin, MyoD1, desmin, and muscle specific actin were employed on a large series of cases (n = 19) of formalin fixed, paraffin-embedded alveolar soft part sarcoma. RESULTS: Minimal scattered nuclear staining was seen with myogenin. All cases had pronounced, nonspecific granular cytoplasmic immunostaining with MyoD1; nuclei were negative. All tumors were negative for desmin and muscle-specific actin. Ultrastructural study in 10 cases failed to reveal features of skeletal muscle differentiation. CONCLUSIONS: Cytoplasmic staining with MyoD1 in alveolar soft part sarcoma may correspond to cross-reactivity with an undetermined cytoplasmic antigen. The lack of immunostaining with myogenin, MyoD1, desmin, and muscle-specific actin provides evidence against a myogenic origin for alveolar soft part sarcoma. PMID- 10383803 TI - An examination of ethical issues raised in the pretreatment of normal volunteer granulocyte donors with granulocyte colony-stimulating factor. AB - OBJECTIVE: To explore some of the ethical issues surrounding the administration of granulocyte colony-stimulating factor (G-CSF) to healthy individuals for the purpose of retrieval of granulocytes. DESIGN: Review of the historical precedent of drug administration to normal blood donors and review of the literature concerning the side effects of G-CSF administration to healthy individuals, particularly as related to granulocyte collection. We identify and discuss some of the ethical questions regarding this issue. RESULTS: Although the short-term side effects of G-CSF use in normal donors are generally felt to be benign, little is known about the long-term side effects. Ethical questions regarding the administration of this drug to normal donors for the purpose of collecting large numbers of granulocytes include the following: Does the potential benefit to a patient/recipient justify the unknown risks to the medicated granulocyte donor? Who should act as an advocate for donors so that their best interests are protected? What is the role and quality of informed consent for donors undergoing G-CSF administration? Is monetary compensation appropriate for donors administered G-CSF as part of a research protocol? CONCLUSIONS: We recommend the establishment of a donor registry to collect the needed data on the side effects of G-CSF on normal donors. Until adequate data are collected, the use of G-CSF and similar agents in normal donors should be regarded as experimental and subject to review by institutional review boards. PMID- 10383804 TI - Influence of etoposide on the retention of radiolabeled low-density lipoprotein in the arterial wall. AB - BACKGROUND: Cardiovascular toxicity of chemotherapy for testicular cancer is a matter of discussion, since highly efficient agents can achieve cure of the disease, even in the metastasized setting. Acute ischemic events during the treatment period and a persistently elevated serum cholesterol thereafter are observations of particular concern in these patients, and the underlying basic mechanisms are unknown to date. OBJECTIVE: To evaluate etoposide, which is part of the standard treatment for testicular cancer, as a potential cause of atherogenesis in an experimental model. SETTING: Aortic 125I-labeled low-density lipoprotein retention was studied in 72 cholesterol-fed rabbits under etoposide treatment and was quantified according to the morphologically assessed type of surface lining. Aortic cholesterol content was determined both by Sudan III staining and quantitatively by a biochemical assay. RESULTS: A reduced uptake of 125I-labeled low-density lipoprotein in the arterial wall was observed in the etoposide-treated animals, which resulted in a size reduction of sudanophilic areas and cholesterol content. Whereas the breakdown of 125I-labeled low-density lipoprotein in the liver was not significantly enhanced, the plasma decay of 125I labeled low-density lipoprotein was faster and serum cholesterol was lower in the etoposide group than in controls. CONCLUSION: Unexpectedly, we found an improvement of arterial wall lipid metabolism under etoposide treatment and can thus exclude this substance as a promoter of atherogenesis in this model. PMID- 10383805 TI - Neuropathology of human immunodeficiency virus 1 infection. Significance of studying in forensic autopsy cases at Dar es Salaam, Tanzania. AB - OBJECTIVE: In sub-Saharan Africa, only a few studies of neurologic complications of human immunodeficiency virus 1 (HIV-1) infection have been done. The authors studied neuropathology of HIV-1 infection in Tanzania. DESIGN: Forensic autopsy study at Dar es Salaam, Tanzania. SETTING: A joint research project between Dar es Salaam, Tanzania, and Kumamoto, Japan. PATIENTS: Thirty patients with risk factors for HIV-1 infection. MAIN OUTCOME MEASURES: Human immunodeficiency virus 1 infection was evaluated by HIV-1 antibody test on postmortem serum samples. The brains of HIV-1-infected persons were studied histopathologically. RESULTS: Infection with HIV-1 was identified on postmortem serum samples in 10 of 30 forensic autopsy cases. Neuropathologic changes of the brain were observed in 8 of the 10 HIV-1-infected persons; these changes consisted of lymphocytic meningitis, bacterial meningoencephalitis, cryptococcal meningoencephalitis, tuberculous meningitis with brain abscesses, and intracerebral hemorrhage. CONCLUSIONS: Because none of the persons studied was suspected to have had brain diseases before autopsy, the results suggest that brain diseases of HIV-1 infected patients are likely to go unrecognized in Tanzania. In addition, the high incidence of neuropathologic findings in HIV-1-infected persons indicates that HIV-1-related brain diseases are common in Tanzania, as they are in developed countries. Further forensic autopsy study will determine the range and prevalence of brain complications and have immediate impact on the management of HIV-1-infected patients in Tanzania and other developing countries. PMID- 10383806 TI - Morphology and other prognostic factors of hepatocellular carcinoma. AB - OBJECTIVE: Hepatocellular carcinoma is a malignancy found worldwide that has typically poor prognosis despite treatment. Although several studies have dealt with prognostic factors, just a few detailed analyses of large series correlating the pathology of hepatocellular carcinoma with prognosis are available. The present study was undertaken to address this limitation. PATIENTS AND METHODS: Our prior clinical study described 432 patients, but sufficient tissue was available for evaluation in only 299 patients. Of these, 224 samples contained primary hepatocellular carcinoma, while the remainder contained only metastatic tumor. Characteristics evaluated included degree of tumor differentiation, associated cirrhosis or hepatitis, presence of cytoplasmic inclusion bodies, and blood vessel invasion by the neoplasm. RESULTS: Of the 224 patients, 71% were male, 65% white, and 73% over the age of 45 years. Ninety-one percent were from North America. A total of 42 patients were found to have cirrhosis. Thirty-five percent had cytoplasmic inclusion bodies, and 25% showed evidence of blood vessel invasion. Tumor response rates (tumor shrinkage) were low (8%) regardless of treatment. Presence of cytoplasmic eosinophilic inclusion bodies and blood vessel invasion were not associated with increased survival. Some histopathologies (pelioid, spindle cell, fibrolamellar) were associated with a better prognosis. Patients with a predominant trabecular pattern (43%) did particularly poorly. Although sex was significantly associated with survival using a univariate analysis, this effect disappeared in a multivariate Cox model that adjusted simultaneously for other factors. CONCLUSION: This investigation suggests that histologic subtype and clinical features may provide useful prognostic information in hepatocellular carcinoma. Poorer survival was observed in males, older patients with poorly differentiated tumors, or when associated with cirrhosis. Age younger than 45 years was a good prognostic factor, and presence of cirrhosis had an adverse effect on survival. PMID- 10383807 TI - Histologic sectioning produces TUNEL reactivity. A potential cause of false positive staining. AB - OBJECTIVE: To determine if the DNA strand breaks caused by tissue sectioning result in terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling (TUNEL) reactivity. METHODS: The incidence and location of TUNEL-positive nuclei were determined in 5- and 15-micron sections of human stomach. Five- and 15-micron sections of tonsil were stained as a positive control. RESULTS: In 5-micron gastric sections, 69% of nuclei were labeled; in 15 micron sections, only 30% were labeled. In the latter sections, almost all labeled nuclei were located at the cut surface of sections. Labeled nuclei did not have apoptotic morphology. Apototic bodies and tingible body macrophages were labeled throughout 15-micron sections of tonsil. CONCLUSIONS: Tissue sectioning creates TUNEL reactivity. The morphologic findings on routine stains should be considered the gold standard for the detection of apoptosis on tissue sections. PMID- 10383809 TI - Atypical cellular chorangioma. AB - We describe an unusual, large atypical cellular chorangioma with abundant mitoses and focal necrosis. Other than premature birth, the prenatal and postpartum clinical course was unremarkable for both the mother and baby. Our case and a few similar cases reported in literature suggest that atypical cellular chorangioma is a benign tumor, despite its worrisome histopathologic features. PMID- 10383808 TI - Axillary sentinel lymph node examination in breast carcinoma. AB - OBJECTIVE: To evaluate whether the type of pathologic examination of breast sentinel nodes (frozen section, step sections, and immunoperoxidase staining) results in different percentages of nodes positive for metastatic disease. DESIGN: Twenty-eight consecutive patients with breast sentinel node biopsies were evaluated by step-sectioning the sentinel node(s) along with performing immunoperoxidase stains for low-molecular-weight cytokeratin and epithelial membrane antigen. SETTING AND PARTICIPANTS: The patients were from a university hospital and large private hospital. MAIN OUTCOME MEASURES: The results of the step sections and immunoperoxidase stains were compared with routine examination, that is, intraoperative frozen section along with a single hematoxylin-eosin slide. RESULTS: Nine cases were positive by routine evaluation, 10 by step sections, and 11 by immunoperoxidase staining. CONCLUSIONS: The large, multi institutional studies of sentinel node utility must take into account the surgical pathology methods used to evaluate these specimens so that uniform techniques, which reliably predict the status of the axillary nodes, can be instituted at all institutions that use this procedure. PMID- 10383810 TI - Complex vascular lesions at autopsy in a patient with phentermine-fenfluramine use and rapidly progressing pulmonary hypertension. AB - Anorectic agents, such as aminorex and fenfluramine derivatives, have been associated previously with the development of primary pulmonary hypertension. The combination diet drug phentermine-fenfluramine (or "phen-fen") was recently marketed in the United States. We describe a case of a 39-year-old woman with rapidly progressing, fatal pulmonary hypertension following administration of phentermine-fenfluramine. Autopsy was remarkable for complex pulmonary vascular lesions most consistent with thrombotic arteriopathy. PMID- 10383813 TI - Diversion colitis: new light through old windows PMID- 10383811 TI - Pathologic quiz case. A 22-year-old man with an abdominal mass. PMID- 10383814 TI - Local sequence alignments with monotonic gap penalties. AB - MOTIVATION: Sequence alignments obtained using affine gap penalties are not always biologically correct, because the insertion of long gaps is over penalised. There is a need for an efficient algorithm which can find local alignments using non-linear gap penalties. RESULTS: A dynamic programming algorithm is described which computes optimal local sequence alignments for arbitrary, monotonically increasing gap penalties, i.e. where the cost g(k) of inserting a gap of k symbols is such that g(k) >/= g(k-1). The running time of the algorithm is dependent on the scoring scheme; if the expected score of an alignment between random, unrelated sequences of lengths m, n is proportional to log mn, then with one exception, the algorithm has expected running time O(mn). Elsewhere, the running time is no greater than O(mn(m+n)). Optimisations are described which appear to reduce the worst-case run-time to O(mn) in many cases. We show how using a non-affine gap penalty can dramatically increase the probability of detecting a similarity containing a long gap. AVAILABILITY: The source code is available to academic collaborators under licence. PMID- 10383815 TI - Fas modulates both positive and negative selection of thymocytes. AB - We studied the functional role of Fas (CD95) in thymic T cell development using the TCR transgenic mice homozygous for the lpr mutation, DO10 lpr/lpr mice. In DO10 lpr/lpr mice, the differentiation of CD4(+)CD8(+) double-positive (DP) thymocytes to CD4(+) single-positive (SP) thymocytes was markedly impaired, as indicated by decreased generation of CD4(+) SP thymocytes and reduced ratio of CD4(+) SP thymocytes to DP thymocytes in lpr/lpr mice compared with those of +/+ mice. Activation of DP thymocytes in the process of positive selection was also significantly inhibited in DO10 lpr/lpr mice, as shown by the lower levels of CD69 expression on DP thymocytes in lpr/lpr mice compared to +/+ mice. Furthermore, the deletion of DP thymocytes induced by in vivo administration of OVA peptide (up to 150 micrograms) and anti-TCR clonotype mAb did not occur in DO10 lpr/lpr mice, whereas these treatments significantly decreased DP thymocytes in DO10 +/+ mice. On the other hand, no significant difference in DO10 transgenic TCR expression on DP thymocytes was found between DO10 lpr/lpr and +/+ mice. Together, these results indicate that Fas is importantly involved in both positive and negative selection of thymocytes. PMID- 10383816 TI - Microfilament assembly is involved in B-cell apoptosis. AB - Crosslinking of the B-cell antigen receptor (BCR) initiates a chain of reactions which culminate in a number of biologic responses, including entry into the cell cycle or cell death. The signals and processes which lead to cell death are slowly being unraveled. Based on the dramatic changes in cell shape which occur during progression of the apoptotic response, activation of cytoskeletal assembly may be critical as this appears to be essential to the mitogenic response. In this study, we demonstrate that crosslinking of the human BCR with anti-IgM antibodies results in the rapid assembly of actin. Our data also suggest that this conversion of G- to F-actin may be a prerequisite for the apoptosis response, as prevention of this conversion by botulinum C2 toxin or cytochalasin D results in rescue of the cells from apoptosis. Prevention of tyrosine kinase activation, disruption of microfilament assembly, and rescue of B lymphocytes from apoptosis imply that tyrosine phosphorylation is needed for both microfilament assembly and apoptosis. In each instance where microfilament assembly is inhibited, anti-IgM-induced activation of the protease CPP32 (caspase) is also inhibited. Taken together, these results suggest that the microfilament system is actively involved in delivering signals for apoptosis. PMID- 10383817 TI - Presentation of MUC1 tumor antigen by class I MHC and CTL function correlate with the glycosylation state of the protein taken Up by dendritic cells. AB - We previously reported that the glycosylated MUC1 tumor antigen circulating as soluble protein in patients' serum is not processed by dendritic cells and does not elicit MHC-Class II-restricted T helper responses in vitro. In contrast, a long synthetic peptide from the MUC1 tandem repeat region is presented by Class II molecules, resulting in the initiation of T helper cell responses. Here we addressed the ability of dendritic cells to present various glycosylated or not glycosylated forms of MUC1 by MHC Class I. We found that three different forms of MUC1, ranging from glycosylated and underglycosylated protein to unglycosylated synthetic peptide, were able to elicit MUC1-specific, Class-I-restricted CTL responses. The efficiency of processing and the resulting strength of CTL activity were inversely correlated with the degree of glycosylation of the antigen. Furthermore, the more efficiently processed 100mer peptide primed a broader repertoire of CTL than the glycosylated protein. PMID- 10383818 TI - Single-cell analysis of costimulation by B cells, endothelial cells, and fibroblasts demonstrates heterogeneity in responses of CD4(+) memory T cells. AB - Human endothelial cells (EC) express MHC class II molecules in vivo and are likely to be involved in presentation of antigens to CD4(+) T cells. We examined, at the single-cell level, EC presentation of superantigens to resting CD4(+) memory T cells. Within 2 h of adherence to class II+ EC early T cell activation is evidenced by translocation of nuclear factor of activated T cells (NFAT), surface expression of CD69, and synthesis of IFN-gamma and IL-2. Naive T cells are not activated. T cell activation is dependent on the prior induction of MHC class II molecules on EC and is blocked by antibodies to LFA-3 (CD58). Our data place EC along a spectrum of antigen-presenting ability. Activated B cells and macrophages trigger more cells to express cytokines than do EC and at lower antigen concentrations; EC are in turn, superior to fibroblasts or smooth muscle cells. Furthermore, the concept of activation thresholds for cytokine synthesis within T cells also extends to earlier activation events: NFAT translocation is relatively easy to trigger, as is CD69 expression; fewer cells can be triggered to express IFN-gamma and fewer still to express IL-2. EC may, therefore, contribute to a graded immune response by inducing qualitatively and quantitatively different responses than professional APC. PMID- 10383819 TI - B7-1 overexpression by thymic epithelial cells results in transient and long lasting effects on thymocytes and peripheral T helper cells but does not result in immunodeficiency. AB - The B7-1 (CD80) molecule provides costimulatory function for the activation of T helper lymphocytes upon encounter with antigen. To investigate the role of this molecule in thymocyte maturation, we have generated transgenic (Tg) mice in which CD80 expression is driven by the keratin 14 promoter (K14). This overexpression of CD80 resulted in the loss of detectable cell surface CD28 expression on thymocytes and a significant reduction in both the surface T cell receptor expression and the ratio of CD4(+) to CD8(+) single-positive thymocytes in Tg animals compared to nontransgenic (non-Tg) controls. While many of these defects were transient, the significant decrease in CD4(+) versus CD8(+) T cell ratio persisted peripherally. Peripheral T cells from these Tg mice were found to be significantly hyporesponsive to T cell mitogens and in mixed leukocyte reaction, effects that our data indicate are due to reduced IL-2 production by Tg T cells upon activation. Despite these functional defects, immunization with both complex and simple protein antigens produced no differences in the proliferative or humoral responses to these antigens between Tg and non-Tg groups. These data indicate that thymic CD80 signaling results in the deletion of significant numbers of CD4(+) T cells but does not culminate in antigen-specific immunodeficiency. PMID- 10383820 TI - The role of B cells in the establishment of T cell response in mice infected with an intracellular bacteria, Listeria monocytogenes. AB - To clarify the role of B cells in the establishment of T cell response against intracellular bacteria, B-cell-deficient (muMT-/-) mice were infected with an intracellular bacteria, Listeria monocytogenes, and T cell response against the bacteria was analyzed. On day 6 of primary Listeria infection, spleen T cells of the muMT-/- mice showed significantly lower levels of proliferative response and IFN-gamma production than those of normal infected mice after in vitro stimulation with listerial antigen. Even in the secondary Listeria infection after immunization with viable bacteria, spleen T cells of the muMT-/- mice proliferated and produced IFN-gamma against listerial antigen at significantly lower levels than those of normal immunized mice. These results demonstrate participation of B cells in priming of Listeria-specific T cells in vivo. However, B cells failed to present Listeria antigen to Listeria-specific T cells in vitro unless Listeria antigen was solubilized. Furthermore, transfer of immune serum from Listeria-infected normal mice failed to enhance the Listeria-specific T cell response of muMT-/- mice. The results indicate that B cells support the T cell response against intracellular bacteria through a mechanism other than their Ig production or antigen presentation function. PMID- 10383821 TI - Expression and potential role of inducible nitric oxide synthase in the central nervous system of Theiler's murine encephalomyelitis virus-induced demyelinating disease. AB - Intracerebral inoculation of susceptible strains of mice with Theiler's murine encephalomyelitis virus (TMEV) results in immune-mediated demyelinating disease. We examined the pathogenic roles of nitric oxide (NO) and inducible NO synthase (iNOS) in TMEV-induced demyelinating disease (TMEV-IDD). The presence of iNOS was confirmed in the spinal cords of TMEV-infected mice using immunohistochemical staining with anti-iNOS antibody on day 0 (control) and days 15, 30, 60, and 120. Aminoguanidine (AG), a specific inhibitor of iNOS, was injected intraperitoneally (ip) on 1, 3, 5, 8, 10, and 12 days post-TMEV inoculation as induction phase or 15, 17, 19, 22, 24, and 26 days as effector phase. Control animals in each experiment received phosphate-buffered saline (PBS) ip at similar time intervals. Few iNOS-positive cells were observed in the spinal cords of naive SJL/J mice. In the early phase (day 15) of TMEV-IDD, an increase of iNOS-positive cells was detected in the leptomeninges and perivascular space of the spinal cords. The number of iNOS-positive cells was increased and reached its peak on day 60, when histology of the animals showed peak infiltration with inflammatory cells. The clinical course of TMEV-IDD on each day postintracerebral infection was significantly reduced in mice treated with AG in the effector phase, and there was no significant difference between mice treated with AG in induction phase versus those administered PBS. Thus, NO production via iNOS appears to be a pathogenic factor in the effector phase of TMEV-IDD. PMID- 10383822 TI - Immune modulation by IL-10 gene transfer via viral vector and plasmid DNA: implication for gene therapy. AB - In the present report, we have evaluated and compared the modulatory effect of the cytokine interleukin (IL)-10 expressed via viral vector or plasmid DNA on viral antigen-induced cutaneous inflammatory lesions. Our data demonstrate the superior potency of both recombinant vaccinia virus and herpes simplex virus IL 10 expression vectors after single intramuscular administration, but the effects were only short term and only functioned in animals lacking immunity to the viral vectors used for modulation. In contrast, modulatory effects achieved by plasmid DNA expressing IL-10 were delayed in onset and milder in effect but were far more persistent than those achieved by viral vectors. Moreover, plasmid DNA expressing IL-10 provided effective modulation when given repeatedly to animals. Our data also showed that IL-10 gene delivery resulted in a systemic and durable modulatory effect while the effect caused by a single IL-10 protein treatment was transient and confined to the injected site. Our results imply that the viral vector system is superior for obtaining short-term effects, whereas the plasmid DNA approach represents a better strategy to achieve gene therapy to modulate chronic inflammatory lesions. PMID- 10383823 TI - Modulation of perforin, granzyme A, and granzyme B in murine natural killer (NK), IL2 stimulated NK, and lymphokine-activated killer cells by alcohol consumption. AB - Alcohol consumption in mice suppresses the cytolytic activity of natural killer (NK) and lymphokine-activated killer (LAK) cells through unknown mechanisms. Herein, we found that alcohol consumption decreased target cell-induced release of granzyme A activity in freshly isolated splenic NK cells, in NK cells stimulated for 18 h with 1000 IU/ml of interleukin 2, and in LAK cells. The total activity and protein expression of granzymes A and B also were lower in these cells than in cells isolated from water-drinking mice. Interleukin 2 increased granzyme A protein expression independent of alcohol consumption; however, this increase was associated with decreased enzyme activity. In contrast, granzyme B protein expression and enzymatic activity increased in response to interleukin 2. Perforin activity and protein expression were reduced in LAK cells generated from alcohol-consuming mice. We conclude that the mechanism underlying the suppression of NK and LAK cytolytic activity by alcohol consumption involves the collective reduction of target-induced release, activity, and expression of perforin and granular proteases. PMID- 10383824 TI - Paul Ehrlich's passion: the origins of his receptor immunology. AB - In 1897, Paul Ehrlich published a selection theory of antibody formation that anticipated the theories of Jerne and Burnet by some 60 years. Ehrlich introduced into immunology the concept of the interaction of physiologically active substances with specific receptors, an idea that still dominates modern immunological thought. In this paper, we point out how Ehrlich's concept matured over 20 years, while it governed his studies in histological staining, in cell physiology, in hematology, and finally in his major contributions in experimental immunology. PMID- 10383825 TI - Stomatin, a MEC-2 like protein, is expressed by mammalian sensory neurons. AB - The molecular mechanism whereby vertebrate primary sensory neurons convert mechanical energy at their receptive fields into action potentials is unknown. In recent years, genetic screens for touch insensitive mutants of the nematode worm Caenorhabditis elegans have led to the identification of several genes required for mechanical sensitivity. A model has been proposed in which a mechanically gated ion channel is connected both to the extracellular matrix and to the cytoskeleton. Displacement of the membrane is proposed to produce a shearing force that pulls the channel open. MEC-2 is thought to play an important role in this complex by linking the ion channel to the cytoskeleton. MEC-2 is highly homologous to a vertebrate protein called stomatin. Stomatin was first isolated from erythrocytes where it is a major integral membrane protein. To date, however, no data on neuronal expression of stomatin in the peripheral nervous system (PNS) or central nervous system (CNS) is available. Here, we have used RT PCR, in situ hybridization, Northern blotting, and immunocytochemistry to demonstrate that stomatin is expressed by all sensory neurons in mouse dorsal root ganglia. Indirect immunofluorescence together with transfection of cultured adult sensory neurons with epitope-tagged stomatin show that stomatin is localized in spots on somatic and axonal membranes. During development, stomatin begins to be expressed by sensory neurons only as target innervation occurs. The onset of expression of stomatin thus coincides with the onset of functional mechanical sensitivity. Together, our data suggest that stomatin, like the C. elegans MEC-2 gene, is expressed in an appropriate temporal and spatial manner to participate in a putative vertebrate mechanotransduction complex. PMID- 10383826 TI - Rapid phosphorylation of Elk-1 transcription factor and activation of MAP kinase signal transduction pathways in response to visual stimulation. AB - The AP-1 transcription factor, which is composed of various combinations of Fos and Jun proteins, is believed to be a key participant in molecular processes that guide activity-dependent changes in gene expression. In this study, we investigated the activity of different MAP kinases that have been implicated in AP-1 activation. We examined the activities of ERK, JNK/SAPK, and p38 MAPK along with their nuclear targets (Elk-1 and c-Jun) in rat visual cortex after light stimulation. The transcription factor Elk-1 (a possible regulator of c-fos expression) was found to be transiently modified by phosphorylation when visual stimulation was applied after a period of dark rearing. In vitro kinase assay with Elk-1 as substrate showed that light stimulation activated MAPK/ERK in visual cortex but not frontal cortex. Furthermore, ERK activation was temporally matched to onset of Elk-1 phosphorylation. The activity of JNK1 (c-Jun N-terminal kinase 1) was elevated at 2-6 h after visual exposure and was also temporally correlated to increase of endogenous P-c-Jun levels and its appearance within the AP-1 DNA-binding complex. The activities of p38 MAP kinases did not change significantly. These results demonstrate the differential engagement of MAPK signaling pathways following sensory stimulation and their relative effects upon AP-1 expression in the intact brain. PMID- 10383827 TI - A cysteine-rich form of Xenopus neuregulin induces the expression of acetylcholine receptors in cultured myotubes. AB - Neuregulin-1 (NRG-1) has diverse functions in neural development, and one of them is to up regulate the expression of acetylcholine receptors (AChRs) at muscle fibers during the formation of neuromuscular junctions. NRG-1 has two prominent alternative splicing sites at the N-terminus; it could be an immunoglobulin (Ig) like domain named Ig-NRG-1 or an apolar cysteine-rich domain (CRD) named CRD-NRG 1. cDNAs encoding Xenopus CRD-NRG-1 were isolated by cross-hybridization with Xenopus Ig-NRG-1 cDNA fragment. The amino acid sequence of Xenopus CRD-NRG-1 is 45 to 70% identical to the human, rat, and chick homologs. Similar to Ig-NRG-1, two variation sites within CRD-NRG-1 were identified at the spacer domain with 0 or 43 amino acids inserted and at the C-terminus of the EGF-like domain to derive either alpha or beta isoform. Two transcripts encoding CRD-NRG-1, approximately 7.5 and approximately 9.0 kb, were revealed in adult brain and spinal cord, but the expression in muscle was below the detectable level. The recombinant Xenopus CRD-NRG-1 when applied onto cultured myotubes was able to induce the tyrosine phosphorylation of ErbB receptors and the expression of AChR. The AChR-inducing activity of CRD-NRG-1 was precipitated by anti-NRG-1 antibody but not by heparin. In situ hybridization showed a strong expression of CRD-NRG-1 mRNA in developing brain, spinal cord, and myotomal muscles of Xenopus embryo. Similar to the results in other species, both CRD-NRG-1 and Ig-NRG-1 may play a role in the developing Xenopus neuromuscular junctions. PMID- 10383828 TI - Differences and developmental changes in the responsiveness of PNS neurons to GDNF and neurturin. AB - We have studied the ability of GDNF and neurturin to promote the in vitro survival of populations of embryonic chicken parasympathetic, sympathetic, and sensory neurons. We show that these neurons are more responsive to one or other of these factors at particular stages of development. Whereas the parasympathetic neurons are more sensitive to neurturin at late embryonic stages, sympathetic neurons are more sensitive to neurturin at early stages. In contrast, sensory neurons of the nodose ganglion are more sensitive to GDNF throughout embryonic development. Using competitive RT/PCR, we measured the levels of mRNAs encoding GDNF and neurturin receptors in purified neurons. All neurons expressed Ret mRNA, which encodes the common receptor tyrosine kinase for GDNF and neurturin. Neurons that were more sensitive to GDNF expressed higher levels of GFRalpha-1 mRNA than GFRalpha-2 mRNA and neurons that were more sensitive to neurturin expressed higher levels of GFRalpha-2 mRNA than GFRalpha-1 mRNA. These results show that populations of PNS neurons differ markedly in their responsiveness to GDNF and neurturin at certain stages of the development and suggest that these differences are governed in part by the relative levels of expression of members of the GFRalpha family of GPI-linked receptors. PMID- 10383829 TI - A soluble version of the receptor-like protein tyrosine phosphatase kappa stimulates neurite outgrowth via a Grb2/MEK1-dependent signaling cascade. AB - Receptor-like protein tyrosine phosphatase kappa (RPTPkappa) is expressed in the nervous system in a manner consistent with a role in axonal growth and guidance. The extracellular domain of RPTPkappa shares structural features with cell adhesion molecules and can support homophilic adhesion. In the present study we produced a soluble Fc-chimeric protein containing the full extracellular domain of RPTPkappa. Following affinity capture, the RPTPkappa-Fc was shown to promote the aggregation of Covasphere beads, confirming its homophilic binding activity. When added to cultures of cerebellar neurons as a soluble molecule, the RPTPkappa chimera stimulated neurite outgrowth. The neurite outgrowth response was substantially inhibited by a cell-permeable peptide inhibitor of Grb2 and by PD 098059, a drug that has been used to inhibit MEK1 activation in a wide range of cell types. These results demonstrate that RPTPkappa can stimulate neurite outgrowth and provide evidence that this might involve the coupling of Grb2 to a MAPK signal transduction cascade. PMID- 10383830 TI - Endothelial trophic support of neuronal production and recruitment from the adult mammalian subependyma. AB - Vascular endothelial cells are among the first cells that ventricular zone neuroblasts encounter during early development. The ventricular zone cells promote angiogenesis by the invading vasculature, with the release of endothelial mitogens. Yet the feedback support of young neurons by endothelial cells (ECs) has not hitherto been explored. We therefore asked whether ECs might participate in neuronal recruitment, by providing neurotrophic support to newly generated neurons. We used the neurogenic subependymal zone (SZ) of the adult rat forebrain as a model system, because of its well-characterized and relatively homogeneous population of neuronal precursor cells. We found that explants of the adult rat SZ raised on ECs generated more neurons, which survived longer, than explants raised on astrocytes, fibroblasts, or laminin. This endothelial trophic effect was humoral, in that it was also noted in SZ explants raised in noncontiguous coculture with ECs grown on porous inserts. RT-PCR for neurotrophin family members revealed that cultures of both human brain- and umbilical cord-derived ECs produced brain-derived neurotrophic factor (BDNF) mRNA, but no detectable NGF, NT-3, or NT-4 mRNA. ELISA revealed that BDNF protein was secreted by ECs into the medium at >1 ng/ml. The neurotrophic effect of ECs could be replaced by added BDNF, and was blocked by addition of 5 microg/ml trkB-Fc to endothelial-SZ cocultures. Thus, endothelial cells can act as sources of secreted BDNF, through which the capillary microvasculature may act to support neuronal recruitment and survival in the CNS. PMID- 10383831 TI - Scoring of skin rejection in a swine composite tissue allograft model. AB - BACKGROUND: For the first time, we define and correlate visual and histologic grading systems of composite tissue allograft (CTA) skin rejection in a large animal model and determine the utility of these grading systems for early diagnosis and monitoring of rejection. MATERIALS AND METHODS: Sixteen pairs of outbred swine underwent transplant of a forelimb osteomyocutaneous free flap. Group I (n = 6) did not receive immunosuppressive therapy. Group II (n = 10) received oral cyclosporin A, mycophenolate mofetil, and prednisone. The flap was visually inspected and protocol skin biopsies were taken at frequent intervals over a 90-day period. Visual Grades 0 (no rejection) to 4 (severe rejection) were assigned based on skin color, bleeding from biopsy site, and blister formation. Histologic Grades 0 to 4 were assigned based on the degree of vasculitis, folliculitis, dermal inflammation, and epidermal degeneration present. RESULTS: All Group I animals progressively rejected their graft by Day 7. Group II grafts survived from 19 and 90 days; 93% of 115 biopsy specimens were read to be within +/-1 histologic score of their assigned flap visual grade. Visual assessment carried an 8% false positive and 39% false negative rate with regard to biopsy proven rejection. However, 81% of missed rejection specimens were histologic Grade 1. Biopsy, when visually indicated, would detect all rejection episodes when histologically Grade 1 or 2 and still potentially reversible. CONCLUSIONS: Visual scoring of CTA skin serves as a useful tool for initially detecting rejection, but repeated histologic evaluation is necessary for monitoring the subsequent course of the graft. PMID- 10383832 TI - L-arginine inhibits ischemia-reperfusion lung injury in rabbits. AB - BACKGROUND: Recent studies have reported that nitric oxide (NO) acts as a cytoprotective mediator in ischemia-reperfusion (IR) lung injury. We hypothesized that the addition of L-arginine to the perfusate would attenuate the increases in microvascular permeability and pulmonary vascular resistance. MATERIALS AND METHODS: Isolated rabbit lungs were reperfused for 60 min after 120 min warm ischemia. Lung injury was assessed using the fluid filtration coefficient (Kf), pulmonary vasucular resistance (PVR) before ischemia and after reperfusion, and a wet-to-dry lung weight ratio (W/D). RESULTS: The Kf of the control group (without L-arginine) was significantly increased after reperfusion. Lungs perfused with L arginine showed attenuation of the IR-induced increases in Kf and PVR. Addition of Nomega-nitro-L-arginine (L-NA), a NO synthase inhibitor, to the perfusate reduced the beneficial effects of L-arginine. The lungs perfused with dibutyryl cyclic GMP (dbcGMP) showed attenuation of IR-induced increases in Kf and PVR. There were no significant differences in the W/D ratio between these groups. CONCLUSIONS: These results demonstrate that L-arginine has beneficial effects on IR lung injury, perhaps due to enhancement of endothelial cGMP levels. PMID- 10383833 TI - Prostaglandin E1 influences pulsatile preservation characteristics and early graft function in expanded criteria donor kidneys. AB - INTRODUCTION: Unlike simple cold storage, machine preservation allows dynamic assessment and manipulation of the donor organ prior to transplantation. We prospectively compared the effects of five pharmacological agents added to the perfusate during machine preservation of expanded criteria donor (ECD) kidneys in order to (1) describe their influence on perfusion parameters and (2) determine their influence on early graft outcome. METHODS: Two hundred seventy-five consecutive ECD kidneys were preserved in our laboratory between 1/1/94 and 12/31/97 by either machine perfusion (MP) or cold storage (CS). ECD kidneys were defined as those requiring pretransplant biopsy. ECD kidneys were divided by method of preservation and MP kidneys were randomized to receive prostaglandin E1 (MP+PGE1), trifluoperazine (TFP), verapamil (VER), papaverine (PAP), mannitol (MAN), or no intervention during the period of machine perfusion. CS kidneys were randomized to receive PGE1 (CS+PGE1), TFP, VER, PAP, or no intervention. All MP kidneys were preserved by continuous hypothermic pulsatile perfusion using Belzer II solution and perfusion parameters were measured every hour during pulsatile perfusion. All CS kidneys were stored in 1.0 L of University of Wisconsin (UW) solution. RESULTS: The addition of PGE1 to machine perfusate increased renal flow and decreased renal resistance. Moreover, the MP+PGE1-treated group was associated with improved early graft function compared to all other groups. The addition of VER, TFP, PAP, or MAN influenced neither the perfusion characteristics nor the incidence of early graft function in MP kidneys. Similarly, the addition of VER, TFP, or PAP did not influence early graft function in the CS kidneys. The CS+PGE1 group exhibited a significantly lower incidence of early graft function than did the MP+PGE1 group. CONCLUSIONS: PGE1 treatment during machine preservation improves hydrostatic perfusion parameters and reduces the incidence of delayed graft function in ECD kidneys. Moreover, the addition of PGE1, TFP, VER, or PAP to UW does not influence early graft function in the CS kidney. PMID- 10383834 TI - Prevention of hepatic ischemia-reperfusion injury by SOD-DIVEMA conjugate. AB - A protective effect of the SOD (superoxide dismutase)-DIVEMA (divinyl ether and maleic anhydride) conjugate on I-R (ischemia-reperfusion) liver injury was demonstrated. Twenty minutes of normothermic hepatic ischemia was induced by clamping the portal triad of Sprague-Dawley rats. Five minutes before the end of ischemia, SOD, SOD-DIVEMA, or NaCl (0.9%) was given intravenously. Using intravital fluorescence microscopy, hepatic microvascular perfusion was analyzed before ischemia and repeatedly during the 120-min reperfusion period. SOD-DIVEMA significantly restored the sinusoidal perfusion rate (control, 98.0 +/- 0.5; NaCl, 65.5 +/- 7. 7; SOD, 81.5 +/- 8.2; SOD-DIVEMA, 95.8 +/- 0.7%) and reduced the number of leukocytes stagnant in acini (control, 4.4 +/- 0.9; NaCl, 36.6 +/- 6.3; SOD, 27.7 +/- 6.8; SOD-DIVEMA, 12.3 +/- 3.3 cells/lobule) and adherent in postsinusoidal venules (control, 55.0 +/- 24; NaCl, 417 +/- 63; SOD, 253 +/- 58; SOD-DIVEMA, 40.0 +/- 14 cells/mm2). In addition, SOD-DIVEMA maintained postischemic hepatocellular integrity. The SOD-DIVEMA-treated group revealed higher serum SOD enzyme activity compared to the SOD group after 120 min of reperfusion (SOD-DIVEMA, 33.0 +/- 5.9; SOD, 8.6 +/- 3.1 U/ml). The beneficial effect of SOD-DIVEMA was most prominent after 120 min of reperfusion, indicating a longer intravascular half-life of SOD-DIVEMA. PMID- 10383835 TI - Pattern of amplification and overexpression of the eukaryotic initiation factor 4E gene in solid tumor. AB - BACKGROUND: An abundance of eukaryotic initiation factor 4E (eIF4E), a rate limiting initiation component in protein synthesis of mRNAs with long 5'-UTRs, has been shown to upregulate a number of oncogene products. Elevation of eIF4E protein has been detected both in infiltrating ductal carcinoma of the breast (IDCA) and in head and neck squamous cell carcinoma (HNSCC) specimens. We hypothesized that malignant progression in solid tumor is associated with progressive eIF4E gene amplification and protein overexpression in cells sampled from the tumor-free resection margin, transition zone (area adjacent to cancer), and tumor core (center of tumor). MATERIALS AND METHODS: Thirty-eight resected specimens were evaluated: 10 IDCA, 13 HNSCC, and 15 benign specimens from similar sites of noncancer patients. IDCA and HNSCC specimens were divided into three different zones: (1) tumor core, (2) transition zone, and (3) tumor-free zone. Quantitative PCR for the eIF4E gene and Western blots for the eIF4E protein were performed on each of these zones and on the normal controls of benign tissue. Immunohistochemical staining was performed to localize the eIF4E protein overexpression in situ. RESULTS: The eIF4E gene was amplified in the tumor core of both IDCA (3.8 +/- 1.4, P < 0.05, Student's t test) and HNSCC (4.3 +/- 1.4, P < 0.05) specimens. Similarly, the eIF4E protein was overexpressed in the cells from these same sites (17.4 +/- 7.3- and 14.0 +/- 9.7-fold elevation, respectively, P < 0.0001). In the transition zone of IDCA and HNSCC, the degrees of eIF4E gene amplification (4.2 +/- 1.0 and 3.7 +/- 1.2, respectively) and overexpression (4.0 +/- 1.0 and 4.4 +/- 4.6, respectively) were intermediate. In contrast, the eIF4E gene copy number and protein level were near baseline in the tumor-free zone. CONCLUSIONS: Progressive eIF4E gene amplification and protein overexpression were present in cells sampled from the microscopically tumor-free margin to tumor core in surgical specimens of both HNSCC and IDCA. In this study, eIF4E gene amplification and protein overexpression appear to be associated with malignant progression in these two solid tumors. PMID- 10383836 TI - Effects of electrically charged particles in enhancement of rat wound healing. AB - BACKGROUND: Many studies have show that various growth factors enhance wound healing in animal models. However, growth factors are expensive and complex and their wound pharmacokinetics are unknown. The clinical trials with growth factors in the treatment of chronic wounds have produced unsuccessful results. A viable alternative to growth factors may be certain biomaterials such as hydrophilic, carbohydrate beads. MATERIALS AND METHODS: Positively charged, negatively charged, or uncharged beads were applied to paired 6-cm rat incisions. The following key aspects of the wound healing process were examined: wound breaking strength and histological analysis. RESULTS: Our data show that wounds treated with positively charged, DEAE Sephadez beads had a 46-50% (P < 0.001) increase in breaking strength over untreated control wounds. A variety of other positively charged, anion exchange materials also elicited a wound healing response, despite the fact that the positively charged chemical moieties as well as the bead matrix were different. In conjunction with the increase in wound breaking strength, an increase in wound macrophage was observed in wounds treated with anion exchangers (P < 0.01). Negatively charged or uncharged beads showed no significant difference from the untreated controls. CONCLUSION: We conclude from this study that the enhancement of wound healing seen with positively charged beads is due principally to the positive charge on the beads; we postulate that the anion exchange between the positively charged beads and tissue is responsible for this enhancement. PMID- 10383837 TI - A new animal model to study intimal hyperplasia in arteriovenous fistulas. AB - BACKGROUND: Intimal hyperplasia (IH) plays a key role in the failure of arteriovenous (AV) fistulas. The animal models available to study IH in AV fistulas are expensive and do not mimic the development of truly stenotic IH. In this study we examined whether goats are a more suitable model to study IH in AV fistulas. MATERIALS AND METHODS: Thirteen direct and four bridge graft AV fistulas between the carotid artery and the jugular vein of goats were explanted 10 to 195 days after creation. Immunohistochemical staining and morphometric measurements of intima and media were performed in the artery, the vein, the toe, and the heel of the venous anastomosis. Ratios of intimal to medial thickness (Ith/Mth) and area (Ia/Ma) were calculated. RESULTS: IH developed in all goats, mainly at the anastomosis (Ia/Ma = 0.17) and the efferent vein (Ia/Ma = 0.31). The artery was almost free of lesions (Ia/Ma = 0.03). In the efferent vein, Ith/Mth varied between 0.59 and 0.68. In the anastomosis the largest value of Ith/Mth was measured at the suture lines (0.88 and 0.91). Absolute intimal area increased with time. IH contained many vascular smooth muscle cells with a patchy display of desmin positivity, an abundance of smooth muscle cell alpha-actin positivity, and almost complete endothelial cell coverage. Occlusion was due to thrombus formation on the IH. CONCLUSION: A clear intimal hyperplasia developed in AV fistulas in goats at locations comparable to those in humans. Therefore, the AV fistula model in the goat may be seen as an effective model to study IH in hemodialysis AV fistulas. PMID- 10383838 TI - Reduction in vascular responsiveness to adrenomedullin during sepsis. AB - BACKGROUND: Although sepsis is characterized by an early, hyperdynamic phase followed by a late, hypodynamic phase, the mechanism responsible for the transition from the hyperdynamic to the hypodynamic state remains unknown. Since recent studies have shown that adrenomedullin (ADM), a novel potent vasodilatory peptide, is upregulated during sepsis, the aim of this study was to determine whether the reduced vascular responsiveness to ADM is associated with the transition from the hyperdynamic phase to the hypodynamic phase of sepsis. MATERIALS AND METHODS: Adult male Sprague-Dawley rats were subjected to sepsis by cecal ligation and puncture (CLP). At 5 and 10 h (i.e., the hyperdynamic phase of sepsis) or 20 h (the hypodynamic phase) after CLP, the thoracic aorta or small intestine was harvested and preconstricted with norepinephrine. Adrenomedullin (10(-7) M) was applied and the percentage of ADM-induced vascular relaxation in the aortic ring and isolated small intestine was determined. RESULTS: The responsiveness to ADM in the thoracic aorta was not altered at 5-10 h, but decreased significantly at 20 h after CLP. Although ADM-induced relaxation in resistance blood vessels of the small intestine did not change at 5 h, it decreased markedly at 10 and 20 h after the onset of sepsis. CONCLUSIONS: Since the transition from hyperdynamic to hypodynamic sepsis takes place between 10 and 20 h after CLP, it is likely that reduced vascular responsiveness to ADM may be responsible for such an event during the course of polymicrobial sepsis. In view of this, maintenance of vascular ADM responsiveness by pharmacologic agents appears to be a novel approach for preventing or delaying the occurrence of hypodynamic sepsis and septic shock. PMID- 10383839 TI - Recombinant growth hormone's effects on the strength and thickness of radiation injured ileal anastomoses: a rat model. AB - BACKGROUND: Small bowel resections following radiotherapy for gynecologic cancers have resulted in significant rates of morbidity and mortality. The objective of this study was to evaluate the effect of rGH on the breaking strength and thickness of radiation-injured ileal anastomoses in an animal model. MATERIALS AND METHODS: Sprague-Dawley rats were treated with 1800 cGy of pelvic irradiation in a single fractionation. Seventeen weeks following pelvic teletherapy an ileo ileostomy was performed. The rats were randomized to receive 2.0 mg/kg/day of rGH for 7 days or placebo. On the seventh postoperative day a segment of ileum surrounding the anastomosis was resected. The segments were tested for breaking strength or were histologically measured for anastomotic thickness. RESULTS: The ileal anastomotic breaking strength in the rGH group was 181 +/- 8.4 g (mean +/- standard error). The breaking strength of ileal anastomoses in the placebo group was 133 +/- 6.9 g (P < 0.05). The rGH group demonstrated a greater anastomotic thickness (1.65 +/- 0.116 mm) than the placebo group (1.17 +/- 0.113 mm, P < 0.05). Of placebo rats 14.7% developed anastomotic leaks compared to 0% of rGH treated animals (P < 0.05). CONCLUSIONS: RGH increased the ileal anastomotic breaking strength by 36% in radiated rats. The anastomotic leak rate was reduced from 14.7% in the placebo group to 0% in the rGH group. These findings correlated with a 41% increase in the thickness of the anastomotic connective tissue in the rGH group. Clinical investigation in selected patients is warranted. PMID- 10383840 TI - Hepatocyte growth factor concentration in rat bile is affected by hepatic resection volume and external biliary drainage. AB - BACKGROUND/AIMS: Hepatocyte growth factor (HGF) concentrations in bile have been shown to be useful in the early assessment of liver function after hepatectomy. The aim of the present study is to prove the hypothesis that the level of bile HGF is proportional to the regeneration capacity of the liver using a rat model. METHODS: Blood and bile were sampled from rats who underwent 30 or 70% hepatectomy, with or without biliary drainage. HGF concentrations were determined using enzyme-linked immunosorbent assay. RESULTS: Liver regeneration was significantly suppressed after hepatectomy in the rats that underwent external biliary drainage. The bile HGF concentration was positively correlated with the resected liver volume within 24 h of hepatectomy, and HGF levels were markedly increased by external biliary drainage. The postoperative changes in plasma HGF were less dramatic. CONCLUSIONS: HGF appears to play an important role in liver regeneration. Bile HGF concentrations, unlike plasma HGF levels, are a good reflection of the hepatic biosynthesis of this growth factor. Increased concentrations of HGF in bile after external biliary drainage may reflect a compensatory response to the continuous loss of hepatocyte growth factor-rich fluid. PMID- 10383842 TI - Prolongation of rat skin and cardiac allograft survival by low molecular weight heparin. AB - BACKGROUND: We have previously reported that very low doses of low molecular weight heparin compounds (LMWH) inhibit a variety of T-cell-mediated reactions by down-regulation of TNF-alpha production. This study tested the efficacy of LMWH in organ transplantation. METHODS: Skin and heterotopic heart transplantations were performed between recipient Wistar rats and donor BN rats. Two doses of LMWH were given sc, 1 and 20 micrograms, each in three protocols, with day of grafting as Day 0: (A) Daily: -1, 0, 1 ellipsis, (B) Late Weekly: -1, 6, 13 ellipsis, and (C) Early Weekly: -7, 0, 7 ellipsis. Doses and schedules were selected based on efficacy in autoimmune models. Skin graft rejection was defined by complete separation of the graft, and heart transplant rejection was defined as cessation of heartbeat. RESULTS: Treatment with 1 microgram (26.8 +/- 2.0 days) and 20 micrograms (24.5 +/- 2.3 days) of LMWH using the Early Weekly protocol significantly prolonged skin allograft survival compared to controls (17.8 +/- 4.4 days), P < 0.001 for both, whereas other protocols did not. Compared to controls (8.3 +/- 1.4 days), treatment with both 1 and 20 micrograms of LMWH using all three protocols significantly prolonged cardiac allograft survival. The efficacy, however, varied considerably. Increase in graft survival ranged from 18% (1 microgram, Daily, 9.8 +/- 0.7 days, P = 0.02) to more than twofold (20 micrograms, Early Weekly, 20.8 +/- 5.5 days, P < 0.001) according to the dose and schedule of LMWH. CONCLUSIONS: Treatment with very low doses of nonanticoagulant LMWH preparations having anti-TNF-alpha activity significantly prolongs rat skin and cardiac allograft survival in a dose- and schedule-dependent manner. PMID- 10383841 TI - Calcium preconditioning, but not ischemic preconditioning, bypasses the adenosine triphosphate-dependent potassium (KATP) channel. AB - BACKGROUND: Recent evidence has implicated the KATP channel as an important mediator of ischemic preconditioning (IPC). Indeed, patients taking oral sulfonylurea hypoglycemic agents (i.e., KATP channel inhibitors) for treatment of diabetes mellitus are resistant to the otherwise profoundly protective effects of IPC. Unfortunately, many cardiopulmonary bypass patients, who may benefit from IPC, are chronically exposed to these agents. Calcium preconditioning (CPC) is a potent form of similar myocardial protection which may or may not utilize the KATP channel in its mechanism of protection. The purpose of this study was to determine whether CPC may bypass the KATP channel in its mechanism of action. If so, CPC may offer an alternative to IPC in patients chronically exposed to these agents. METHODS: Isolated rat hearts (n = 6-8/group) were perfused (Langendorff) and received KATP channel inhibition (glibenclamide) or saline vehicle 10 min prior to either a CPC or IPC preconditioning stimulus or neither (ischemia and reperfusion, I/R). Hearts were subjected to global warm I/R (20 min/40 min). Postischemic myocardial functional recovery was determined by measuring developed pressure (DP), coronary flow (CF), and compliance (end diastolic pressure, EDP) with a MacLab pressure digitizer. RESULTS: Both CPC and IPC stimuli protected myocardium against postischemic dysfunction (P < 0.05 vs I/R; ANOVA with Bonferroni/Dunn): DP increased from 52 +/- 4 (I/R) to 79 +/- 2 and 83 +/- 4 mmHg; CF increased from 11 +/- 0.7 to 17 +/- 2 and 16 +/- 1 ml/min; and EDP decreased (compliance improved) from 50 +/- 7 to 27 +/- 5 and 31 +/- 7 mmHg. However, KATP channel inhibition abolished protection in hearts preconditioned with IPC (P < 0.05 vs IPC alone), but not in those preconditioned with CPC (P > 0.05 vs CPC alone). CONCLUSIONS: (1) Both IPC and CPC provide similar myocardial protection; (2) IPC and CPC operate via different mechanisms; i.e., IPC utilizes the KATP channel whereas CPC does not; and (3) CPC may offer a means of bypassing the deleterious effects of KATP channel inhibition in diabetic patients chronically exposed to oral sulfonylurea hypoglycemic agents. PMID- 10383843 TI - Effects of deletion-type human hepatocyte growth factor on murine septic model. AB - BACKGROUND: Sepsis is known to be the main cause of multiple organ failure. The liver especially is vulnerable to the stress of infection. In this study, the effects of deletion-type human hepatocyte growth factor (dHGF) on a murine septic model were studied. MATERIALS AND METHODS: Sepsis was induced in male adult Sprague-Dawley rats by cecal ligation and puncture method (CLP). Controls were given a sham operation. Intravenous injection of 1000 micrograms/kg dHGF or the same volume of vehicle was given every 12 h for 3 days before and/or after the CLP from a central vein catheter inserted 1 week prior to the operation. The daily percentage of survival after CLP was followed up for 1 week, and blood samples and liver specimens were collected from the surviving animals 72 h after CLP or sham operation. RESULTS: The survival rate, the degree of liver damage and liver protein synthesis, and coagulation function were all favorable in the dHGF treated animals compared to the untreated animals. Immunohistochemical staining showed that dHGF prevented the disappearance of thrombomodulin (TM) in liver sinusoid endothelium. CONCLUSIONS: dHGF appears to prevent liver injury caused by disturbance of microcirculation through preservation of TM expression and the antithrombotic function in the endothelium of sinusoids. dHGF also facilitates repair of damaged hepatic tissue by stimulating regeneration of the cells and by preserving hepatic functions such as protein synthesis. dHGF exerts protective effects on even quiescent hepatocytes, but is most effective on injured but competent hepatocytes. PMID- 10383844 TI - Diet-induced changes in endothelial-dependent relaxation of the rat aorta. AB - PURPOSE: Hypertension (HTN), hyperlipidemia (HLP), and hyperinsulinemia are known risk factors for the development of cardiovascular disease. Each has independently been shown to be associated with impaired endothelial function, as demonstrated by decreased endothelial derived relaxation (EDR). Previous work in our laboratory has shown that rats fed a high-fat sucrose (HFS) diet will become insulin resistant, hypertriglyceridemic, and hypertensive. We hypothesize that the development of these diet-induced risk factors is associated with endothelial dysfunction and a significant decrease in EDR. Furthermore, the endothelial dysfunction will be improved by returning to a normal (low-fat complex carbohydrate (LFCC)) diet. METHODS: Adult, male Fischer rats were fed either a LFCC or a HFS diet for 6 months (n = 8 in each group). A third group of rats (SWITCH) was fed a HFS diet for 6 months and then changed to a LFCC diet for 4 weeks. Blood pressure was measured via the tail-cuff method weekly. The rats were sacrificed and aortic ring segments were placed in physiologic tissue baths for measurement of vascular reactivity to various agents. Arterial ring segments were constricted with potassium chloride (K) and phenylephrine (PE). Endothelial dependent vasorelaxation was measured with acetylcholine (Ach), bradykinin (BK), and calcium ionophore (CA). Endothelial-independent relaxation was measured using sodium nitroprusside (NTP). RESULTS: The HFS diet group developed HTN compared to LFCC group. Vasoconstriction to K and PE were similar in all groups. Vasorelaxation to Ach, BK, and CA was significantly decreased in the HFS group, but returned to baseline in the diet-switched group, as did the systolic blood pressure. There were no differences in relaxation to NTP. CONCLUSIONS: HFS diet induced HTN is associated with significantly decreased EDR. Switching to a low fat diet reverses this effect. The vascular smooth muscle contraction and endothelial-independent relaxation are not affected by the diet-induced risk factors. There is a direct and reversible effect of an HFS diet on endothelial function and blood pressure. PMID- 10383845 TI - Cold ischemia induces endothelin gene upregulation in the preserved kidney. AB - BACKGROUND: Prolonged cold ischemia time (CIT) can lead to posttransplant renal dysfunction; however, the pathophysiology remains unclear. Endothelin (ET), a potent vasoconstrictive peptide, may play a role in this injury. The purpose of this study was to determine if cold ischemia could induce renal ET-1 gene upregulation and to localize ET-1 peptide expression in the hypothermic kidney. MATERIALS AND METHODS: Kidneys from Lewis rats were perfused with Viaspan, harvested, and stored at 4 degrees C for varying periods of CIT: 0, 6, 24, and 48 h. Preproendothelin-1 (ppET-1) gene upregulation was measured using a reverse transcription polymerase-chain reaction. ET-1 peptide expression was localized using immunohistochemistry. RESULTS: Control kidneys (0 h CIT) had 0. 56 +/- 0.22 DU of ppET-1 mRNA. After 6 h of CIT, a 2.3-fold increase in this level was noted. Following 24 h of CIT, ppET-1 mRNA was significantly upregulated to 1.96 +/- 0.38 DU (P < 0.05). Immunohistochemistry revealed typical vascular ET-1 staining in control kidneys. At 6 h of CIT, a significant increase in the expression of ET-1 was noted in the peritubular capillaries and vasa recta. After 24 h, intense staining for ET-1 was seen in the medullary collecting ducts. After 48 h of CIT, early cellular necrosis was present along with global decreases in ET-1 expression and ppET-1 mRNA levels. CONCLUSIONS: This study demonstrates that 24 h of cold preservation can induce significant upregulation of the renal ET-1 gene and increase expression of the ET-1 peptide localized to both vascular endothelial and tubular epithelial surfaces of the kidney. Consequently, prolonged cold ischemia prior to transplantation may lead to delayed renal function following revascularization via endothelin-induced vasoconstriction and/or tubular impairment. PMID- 10383846 TI - Dexamethasone suppresses vascular smooth muscle cell proliferation. AB - BACKGROUND: Experimental studies in vivo have demonstrated that dexamethasone inhibits neointimal hyperplasia following arterial injury. The mechanisms of this inhibition have not been clearly defined. Our objective was to test the hypothesis that dexamethasone directly suppresses smooth muscle cell (SMC) proliferation by inhibiting cell cycle progression and the expression of key cell cycle-dependent genes. METHODS: Cultured rat aortic SMC were treated with incremental concentrations of dexamethasone and cell number was determined after 72 h. To determine if dexamethasone inhibited cell cycle progression, cells were synchronized, then restimulated to enter the cell cycle, and treated with or without dexamethasone. DNA synthesis was determined 24 h after restimulation by measuring [3H]thymidine incorporation. To define the point of action of dexamethasone in the cell cycle, synchronized SMC were treated with dexamethasone (10(-7) M) at various time points after entry into the cell cycle. Flow cytometry and Northern blots were performed to examine cell cycle progression and the expression of smooth muscle cell cycle-dependent genes c-fos, c-myc, and thymidine kinase (TK). RESULTS: Dexamethasone treatment induced a concentration dependent inhibition of SMC proliferation and DNA synthesis. The cell cycle progression of synchronized SMC from G1 into S phase was inhibited by dexamethasone, even when added as late as 16 h after restimulation. The expression of TK was suppressed by dexamethasone, while c-fos and c-myc were not affected. CONCLUSIONS: Dexamethasone inhibits the proliferation of SMC in a concentration-dependent fashion. This inhibition is associated with a block in cell cycle progression late in G1 phase of the cell cycle. Consistent with this finding, dexamethasone does not alter the expression of the early cell cycle dependent genes c-fos and c-myc, but significantly inhibits the expression of TK, a marker of late G1 phase. PMID- 10383847 TI - Hepatocyte growth factor supply accelerates compensatory hypertrophy caused by portal branch ligation in normal and jaundiced rats. AB - BACKGROUND: Hepatocyte growth factor (HGF), first identified as the most potent mitogen for hepatocytes, significantly stimulates liver regeneration after hepatectomy. In this report, we examined whether HGF is also useful in accelerating compensatory hypertrophy caused by portal branch ligation in normal and jaundiced rats. MATERIALS AND METHODS: Normal and reversible obstructive jaundiced rats underwent portal ligation of the left lateral and median branches, which supply approximately 70% of the total volume of the liver. Simultaneously, the animals were continuously treated with either recombinant human HGF (rhHGF) or vehicle alone via an intraperitoneally implanted osmotic pump. Two and four days after portal ligation, the degree of compensatory hypertrophy in unoccluded lobes was examined by measuring the wet weight ratios of the unoccluded lobes to the whole liver and the 5-bromo-2'-deoxyuridine labeling index of hepatocytes in each group. RESULTS: The HGF treatment significantly increased the wet weight ratios and the DNA synthesis in nonoccluded lobes 2 and 4 days after portal ligation in both normal and jaundiced rats. Moreover, rhHGF supply promptly decreased serum total bilirubin level in jaundiced rats. CONCLUSIONS: Continuous rhHGF administration not only accelerates compensatory hypertrophy in normal and jaundiced rats but also ameliorates hyperbilirubinemia in jaundiced rats. PMID- 10383848 TI - Enhancement of hypothermic heart preservation with fructose 1, 6-diphosphate. AB - BACKGROUND: We hypothesized that the addition of fructose 1, 6-diphosphate (FDP) to a hypothermic heart preservation solution could improve metabolic recovery because it has several beneficial effects. MATERIALS AND METHODS: Twenty adult Sprague-Dawley rats were used to study hypothermic heart preservation. The hearts were removed under general anesthesia and preserved at 4 degrees C in Euro Collins solution (30 ml/kg) for 8 h. In the study group (N = 10), FDP (5 mM) was added to the Euro-Collins solution. In the control group (N = 10), no FDP was added. Heart function was studied after preservation using a working heart model. The ability of various concentrations of fructose 1,6-phosphate to passively diffuse through an egg phosphatidylcholine multilamellar vesicle (MLV) membrane bilayer was examined. RESULTS: Cardiac output ranged from 17.0 +/- 1.9 to 24.9 +/ 1.6 ml/min in the study group vs 2.0 +/- 1.0-12.3 +/- 1.7 ml/min for controls, average aortic flow was 10. 8 +/- 1.4 ml/min in the study group vs -1.3 +/- 1.6 ml/min for controls, and maximum LV generated power was 22.8 +/- 1.7 J/min vs 10.1 +/- 1.6 J/min for controls. Coronary flow, left ventricular stroke volume and stroke work, and myocardial oxygen consumption were much higher in the study group than in the control group. Coronary vascular resistance was lower in the study group than in the control group. Electron microscopic study indicated that many myocytes displayed patches of swollen mitochondria in the control group, but was rarely observed in the study group. The addition of 50 mM FDP caused substantial changes in MLV permeability. No dose of sucrose buffers outside the vesicles resulted in a significant changes of MLV permeability. CONCLUSIONS: Our results indicate that the addition of FDP to Euro-Collins solution significantly improves hypothermic rat heart preservation, and FDP appeared to cross the membrane bilayer. PMID- 10383849 TI - Hindlimb ischemia-reperfusion increases complement deposition and glycolysis. AB - BACKGROUND: Hindlimb ischemia-reperfusion (HIR) impairs cellular energy metabolism and causes local muscle injury possibly through free radical or complement-mediated mechanisms. MATERIALS AND METHODS: To determine the relationship among myocellular energetics, histopathological injury, and mediator activity, male Wistar rats underwent 4 h of Sham (n = 8), Unilateral (n = 8), or Bilateral (n = 8) hindlimb ischemia followed by 4 h of reperfusion. All rats underwent 31P magnetic resonance spectroscopy of their right gastrocnemius muscle to determine various high-energy phosphate ratios including ATP to Pi (ATP/Pi, a measure of energy status) and phosphocreatine to Pi (PCr/Pi, a measure of thermodynamic capacity). Gastrocnemius muscles were then harvested to determine muscle damage and complement membrane attack complex (MAC) deposition by immunohistochemical staining [grade 0 (none) to 3 (very severe)] and to measure glutathione (GSH), DNA, and enzyme activities: beta-hydroxyacyl-CoA dehydrogenase, phosphofructokinase, and citrate synthetase. RESULTS: HIR was associated with significant declines in ATP/Pi and PCr/Pi (P < 0.001). Progressively more severe HIR (Sham, Unilateral, Bilateral) was associated with greater MAC deposition (0. 0 +/- 0.0, 1.0 +/- 0.3, 1.5 +/- 0.4, P = 0.06, mean +/ SEM) and histological damage (0.0 +/- 0.0, 0.9 +/- 0.3, 1.3 +/- 0.4, P < 0. 05). GSH levels, beta-hydroxyacyl-CoA dehydrogenase, and citrate synthetase activities were not affected by HIR, but phosphofructokinase activity increased (24.09 +/- 2.42, 35.16 +/- 5. 26, 59.29 +/- 9.82 mmol/mg of DNA/min, P < 0.05). Although GSH levels were not significantly altered, complement deposition was closely associated with skeletal muscle injury and compensatory changes in glycolysis. Alterations in myocellular bioenergetics after HIR closely paralleled complement deposition rather than GSH depletion. CONCLUSIONS: Therapeutic strategies aimed at controlling complement activity and assessment techniques based on bioenergetics may allow more precise determinations of the effects of HIR injury. PMID- 10383850 TI - Antiendotoxin agents share molecular homology within their lipopolysaccharide binding domains. AB - BACKGROUND: The purpose of this study was to determine whether antiendotoxin agents exhibit molecular homology within their lipopolysaccharide (LPS) binding domains, suggesting a common mechanism of action. We hypothesized that the presence of positively charged basic amino acids or a paucity of negatively charged acidic amino acids, or both, would be a critical characteristic of that portion of the molecule that binds to the highly negatively charged deep core/lipid A (DCLA) region of LPS. MATERIALS AND METHODS: We analyzed the amino acid sequences of the variable light (VL) and heavy (VH) chain complementarity determining regions (CDRs) of anti-DCLA monoclonal antibodies (mAbs) 1B6, 5A5, and 7C5 and compared them with (1) the CDRs of three irrelevant control mAbs and (2) the LPS binding region of bactericidal permeability-increasing protein (BPI). We purified and amplified the specific nucleotide sequences of the variable regions using reverse transcriptase polymerase chain reaction. DNA was sequenced by dideoxy termination, and protein sequences were deduced and analyzed. The percentages of acidic, basic, polar, and hydrophobic amino acids within VH and VL chain CDRs were determined. RESULTS: We identified a paucity of negatively charged acidic amino acids exclusively within VL chain CDRs of anti-DCLA mAbs (P < 0.005). Although increased, the number of positively charged basic residues was not statistically significantly different; neither was the number of polar or hydrophobic amino acids. Conclusions. Our data suggest that the near absence of negatively charged acidic residues is critical for LPS binding. This characteristic appears to reside exclusively in the VL chain CDRs of anti-DCLA mAbs. PMID- 10383851 TI - A reactive oxygen-generating system activates nuclear factor-kappaB and releases tumor necrosis factor-alpha in coronary smooth muscle cells. AB - BACKGROUND: Recently we reported that bacterial lipopolysaccharide (LPS) stimulates release of tumor necrosis factor alpha (TNF-alpha) from porcine coronary arteries and smooth muscle cells cultured from those vessels. It has also been reported that plasma levels of TNF-alpha are elevated after myocardial infarction. Since it is known that the production of reactive oxygen intermediates (ROI) occurs during ischemia and ROI are suggested activators of the nuclear regulatory factor kappaB (NF-kappaB), we tested the hypothesis that release of TNF-alpha from smooth muscle cells could also be stimulated with a ROI generating system. MATERIALS AND METHODS: Smooth muscle cells were isolated from porcine coronary arteries. Confluent cells in 48-well culture dishes were treated for 30 min with 0.003 units/ml xanthine oxidase (XO) and 2 mM hypoxanthine (HX) added to the culture medium. The medium was then removed and the cells were washed three times and fresh medium without HX-XO was added. Then, at 1, 3, and 6 h the medium was removed and analyzed for biologically active TNF-alpha. In other experiments, smooth muscle cells were treated with 20 micrograms/ml LPS for 6 h and aliquots of medium analyzed for TNF-alpha. Untreated cells served as controls. Data were analyzed by two-way ANOVA with repeated measures. Extracts of total cell protein were prepared and activation of NF-kappaB was determined by electrophoretic mobility shift assay. RESULTS: Treatment of cells with HX-XO stimulated release of TNF-alpha, which rose to a maximum of 17.5 +/- 1.7 units/mg cell protein at 6 h. This was significantly higher (P < 0. 05) than release stimulated by LPS (10.2 +/- 1.0 units/mg at 6 h) or TNF-alpha detected in the culture medium from untreated control cells (4.2 +/- 0.9 units/mg protein at 6 h). Both HX/XO and LPS activated NF-kappaB. CONCLUSIONS: These results support the conclusion that coronary smooth muscle cells are a potential source of TNF alpha during events that are associated with formation of ROI such as myocardial ischemia. PMID- 10383852 TI - Normal perinatal rise in serum cholesterol is inhibited by hepatic delivery of adenoviral vector expressing apolipoprotein B mRNA editing enzyme (Apobec1) in rabbits. AB - BACKGROUND: Prenatal or neonatal hepatic gene delivery may result in more effective therapy for inborn errors of metabolism due to the immature immune system of the perinatal animal, and the ability to intervene prior to any significant cellular damage. Newborn New Zealand White rabbits have low serum levels of cholesterol at birth, with a significant and sustained rise of cholesterol while they are nursing. We used this physiologic hypercholesterolemia model to study the effect of adenovirus-mediated hepatic gene transfer of rat apolipoprotein B mRNA editing enzyme (Apobec1) on modulation of plasma cholesterol levels. METHODS AND RESULTS: Transcutaneous injection of recombinant adenovirus expressing Apobec1 (AvApobec1) into the liver of newborn rabbits in vivo resulted in efficient Apobec1 expression until Day 50, as detected by PCR Southern blot analysis. By in vitro editing assay, liver extracts of AvApobec1 treated rabbits were found to have apoB mRNA editing activities of approximately 12, 15, and 15%, on Days 2, 10, and 20 after AvApobec1 administration, compared with 0% editing activity in AvLacZ control vector-injected animals. This physiological level of Apobec1 expression was associated with the production of apoB-48-containing lipoprotein particles from rabbit liver, with a concomitant 30% reduction in total plasma cholesterol compared to AvLacZ-treated or untreated control animals. CONCLUSION: Neonatal intrahepatic delivery of a first-generation adenoviral vector results in efficient gene transfer with little immune response, suggesting that repeated administration may be possible in the neonatal period. PMID- 10383853 TI - Shortened elastase infusion time in the elastase-induced rat aneurysm model. AB - OBJECTIVE: This study was performed to determine whether it would be possible to shorten the elastase infusion time in the elastase-induced rat aneurysm model. METHODS: The abdominal aortas of 76 male Sprague-Dawley rats were dissected and infused for 10, 20, 30, 60, or 120 min with a solution containing 14 U of elastase or for 30 min with saline solution (control). After infusion, the rats were evaluated every day for calculation of the mortality rate. On day 7, the surviving rats underwent laparotomy. The diameter of the aorta was measured, and the aortic tissue was excised for histologic examination. RESULTS: There were no deaths among the rats infused for 10, 20, or 30 min. The mortality rate was 64% in the 60-min and 90% in 120-min groups. There were no significant differences in the diameters of the 30-min saline-infused aortas and the 10- or 20-min elastase infused aortas. The diameter of the 30-min elastase-infused aortas (6.2 +/- 2.1 mm) was significantly larger (P < 0.01) than the diameters of the 10- and 20-min elastase-infused aortas. There were no significant differences between the 30-min and the 60- and 120-min elastase-infused aortas. Microscopically, there was total or subtotal loss of elastic tissue and marked inflammatory cell infiltration of the aortic wall in the 30-, 60-, and 120-min elastase-infused aortas. CONCLUSIONS: It is possible to shorten the elastase infusion time from 120 to 30 min in the elastase-induced rat aneurysm model. This shortening of infusion time reduces the experimental time and mortality rate. PMID- 10383854 TI - The role of ischemic preconditioning in the recruitment of rolling and adherent leukocytes in hepatic venules after ischemia/reperfusion. AB - BACKGROUND: We have recently shown that hepatic ischemia/reperfusion (I/R) results in rolling and adherence of leukocytes in terminal hepatic venules (THV) followed by hepatic enzyme elevation and tissue destruction. The objective of this study was to determine the effect of ischemic preconditioning on the recruitment of leukocytes in THV after liver I/R. METHODS: Left hepatic lobe ischemia was induced for 5 min (preconditioning) in anesthetized C57B1/6 mice followed by reperfusion for 10 min and then prolonged ischemia for 30 min. The number of rolling, saltating, and adherent leukocytes in THV was measured at 0.5, 2, 5, 12, and 24 h after reperfusion using intravital video microscopy. Matching sham groups were evaluated after 30 min of ischemia. RESULTS: Hepatic I/R elicited significant increases in the number of rolling, saltating, and adherent leukocytes, with peak values observed at 30 min and 5 h after reperfusion. All of these responses were significantly attenuated in mice undergoing ischemic preconditioning. Rolling leukocytes in THV following I/R without preconditioning reached peak levels of 25.2 +/- 1.4 leuk/2 min (leukocytes/2 min) at 30 min reperfusion and 31.4 +/- 1.5 leuk/2 min at 5 h reperfusion. With ischemic preconditioning these values fell to 12.3 +/- 0.9 leuk/2 min and 14.4 +/- 1.0 leuk/2 min, respectively (P < 0.001). Similarly, adherent leukocytes in nonpreconditioned mice reached peak values of 4.8 +/- 1.3 leuk/2 min at 30 min reperfusion and 8.3 +/- 1.2 leuk/2 min at 5 h reperfusion compared with 2.0 +/- 1.5 leuk/2 min and 1.6 +/- 1.1 leuk/2 min in preconditioned mice, respectively (P < 0.001). CONCLUSION: Ischemic preconditioning attenuates the initial events leading to leukocyte-mediated hepatic destruction following I/R injury. Delineating these mechanisms may play an important role in hepatic transplantation, resection, shock, and sepsis. PMID- 10383855 TI - Increased expression of insulin-like growth factor-I in serum and liver after recombinant human growth hormone administration in thermally injured rats. AB - BACKGROUND: Recombinant human growth hormone (rhGH) has been shown to modulate the hypermetabolic response and the hepatic acute-phase response after thermal injury. In vitro studies, however, demonstrated that rhGH activates insulin-like growth factor-I (IGF-I) gene transcription and production, suggesting that rhGH may exert some of its effects indirectly through IGF-I stimulation. The purpose of this study was to determine the effects of rhGH on serum and hepatic IGF-I in thermally injured rats. METHODS: Sprague-Dawley rats (56 males) receiving a 60% TBSA third-degree scald burn were randomly divided to receive either rhGH (2.5 mg/kg/day im) or saline (control). Rats were sacrificed on postburn days 1, 2, 5, and 7 and serum IGF-I, hepatic IGF-I mRNA, and IGF-I protein concentration were measured. The physiologic response to changes in IGF-I levels was evaluated by measuring hepatocyte proliferation, total liver protein concentration, and muscle dry/wet weights. RESULTS: Serum IGF-I was increased from postburn day 1 through day 7 in rats receiving rhGH compared to controls (P < 0.05). Hepatic IGF-I mRNA and IGF-I protein expression were increased from day 1 to 7 after burn in animals receiving rhGH when compared to controls (P < 0.05). Recombinant hGH increased hepatocyte proliferation at 5 days and total liver protein concentration at 5 and 7 days postburn compared to controls (P < 0.05). Muscle dry/wet weights increased in rats receiving rhGH at 7 days after burn compared to controls (P < 0.05). SUMMARY: Liver and serum IGF-I levels decreased after a thermal injury. Recombinant hGH attenuated this decrease by stimulating hepatic IGF-I expression. Increases in IGF-I were associated with increases in hepatocyte proliferation and protein concentration in liver and muscle. CONCLUSION: We suggest that rhGH modulates the hypermetabolic response through IGF-I stimulation in the hepatic parenchyma. PMID- 10383856 TI - Glutamine protects against doxorubicin-induced cardiotoxicity. AB - Doxorubicin (DOX) dose-intensive therapy for breast cancer is limited by a cardiomyopathy that often results in overt congestive heart failure. We hypothesized that dietary glutamine (GLN) can diminish DOX-induced cardiotoxicity by maintaining tissue glutathione (GSH) levels and thus preventing the proposed mechanism of cardiac injury: oxidation. METHODS: Forty-two female Fisher 344 rats were randomized into one of six groups: GLN + saline (SAL), GLN + DOX, freamine (FA) + SAL, FA + DOX, H2O + SAL, and H2O + DOX. Rats were pair-fed chow and gavaged with 1 g/kg/day GLN or an isonitrogenous amount of FA or H2O for 28 days. Rats were injected intravenously with a single dose of SAL or 9 mg/kg DOX on day 7 of gavage. At 28 days (21 days post-DOX), rats were sacrificed and blood and cardiac tissue were assayed for GLN and GSH content and lipid peroxidation (LP). RESULTS: There were no differences in cardiac GSH levels and cardiac lipid peroxidation in GLN + SAL versus GLN + DOX groups. However, blood and cardiac GSH levels were significantly decreased in H2O + DOX and FA + DOX groups compared to controls (H2O + SAL and FA + SAL). CONCLUSION: These data suggest that dietary GLN supplementation may diminish DOX-induced oxidative damage and thus cardiotoxicity through upregulation of cardiac GSH metabolism. PMID- 10383857 TI - Multilocus sequence typing: molecular typing of bacterial pathogens in an era of rapid DNA sequencing and the internet. AB - Multilocus sequence typing is a development of multilocus enzyme electrophoresis in which the alleles at multiple house-keeping loci are assigned directly by nucleotide sequencing, rather than indirectly from the electrophoretic mobilities of their gene products. A major advantage of this approach is that sequence data are unambiguous and electronically portable, allowing molecular typing of bacterial pathogens (or other infectious agents) via the Internet. PMID- 10383858 TI - The role of short sequence repeats in epidemiologic typing. AB - Short sequence repeats (SSRs), also known as variable number of tandem repeats or micro-satellites, are inherently unstable entities that undergo frequent variation in the number of repeated units through slipped strand mispairing during DNA synthesis. In humans, unit number variability in SSRs has been associated with the occurrence of specific genetic diseases, whereas in micro organisms SSRs have been elegantly linked to modulation of gene expression. Knowledge of the functional constraints imposed upon the SSRs sheds light on their potential use as molecular clocks for monitoring microbial genome evolution. Although microbial SSR genotypes have been used with increasing frequency for studying the epidemiology and evolution of microbial strains and isolates, such approaches should be used with caution. PMID- 10383859 TI - Biocatalysts for clean industrial products and processes. AB - Biocatalysis inherently offers the prospect of clean industrial processing and has become an accepted technology throughout most sectors. The convergence of biology and chemistry has enabled a plethora of industrial opportunities to be targeted, while discoveries in biodiversity and the impact of molecular biology and computational science are extending the range of natural and engineered biocatalysts that can be customised for clean industrial requirements. PMID- 10383860 TI - Photorhabdus toxins: novel biological insecticides. AB - Following concerns over the potential for insect resistance to insecticidal Bacillus thuringiensis toxins expressed in transgenic plants, there has been recent interest in novel biological insecticides. Over the past year there has been considerable progress in the cloning of several alternative toxin genes from the bacteria Photorhabdus luminescens and Xenorhabdus nematophilus. These genes encode large insecticidal toxin complexes with little homology to other known toxins. PMID- 10383861 TI - Biofilms. AB - Outside of the laboratory, most microbes grow as organised biofilm communities on surfaces. The past year has seen important advances in our understanding of how cells initiate biofilm formation. We have also begun to appreciate how cells can co-ordinate their activities and build the complex structures of mature biofilms that afford protection for their inhabitants. PMID- 10383862 TI - Ribosomal DNA sequencing as a tool for identification of bacterial pathogens. AB - Conventional methods for the identification and characterization of clinical isolates of bacterial pathogens sometimes fall short when such isolates exhibit unusual phenotypic profiles. Recent advances in DNA sequencing technology have greatly enhanced the ability of the microbiologist to determine the identity of a bacterial isolate. Given the relative objectivity of DNA sequence information and growing availability of sequence information databases, a significant movement is now afoot to use molecular methods for the identification of clinical pathogens. PMID- 10383863 TI - Thermophilic and halophilic extremophiles. AB - The microbiology of extremely hot or saline habitats is a fast moving field with many new successes in the enrichment and isolation of new organisms and in an understanding of molecular factors that impart stability on thermostable and halophilic biomolecules. The results of these studies have shed new light on our understanding of prokaryotic diversity and structural biochemistry. PMID- 10383864 TI - Terminal restriction fragment length polymorphism (T-RFLP): an emerging method for characterizing diversity among homologous populations of amplification products. AB - Terminal restriction fragment length polymorphism is a recent molecular approach that can assess subtle genetic differences between strains as well as provide insight into the structure and function of microbial communities. The technique has both high sensitivity and throughput making it ideal for comparative analyses. PMID- 10383865 TI - Commodity scale production of sugars from starches. AB - The production of sugars from starch sources is an industry that exists in its present form due to the application of industrial enzymology to solve process related problems. As the industry matures, the demand for more efficient enzymes leading to higher quality products and lower production costs for the starch processor has increased. Researchers are now finding or tailoring enzymes for specific operational needs of the processor using a combination of tools such as protein engineering, directed evolution and improved accessing of natural diversity. PMID- 10383866 TI - Novel approaches for discovering industrial enzymes. AB - New technologies for enzyme discovery are changing the rules of the game for industrial biocatalysis. More kinds of enzymes are available, their hardiness is increasing, and their costs are coming down. These changes are the key drivers for a rebirth of interest in industrial applications of enzymes. The major enabling discovery approaches include screening of biodiversity, genomic sequencing, directed evolution and phage display. PMID- 10383867 TI - Gene expression systems for lactic acid bacteria. AB - Considerable advances have been made in the genetics and molecular biology of lactic acid bacteria, including Lactococcus, Lactobacillus, Leuconostoc, Pediococcus and Streptococcus spp. These have resulted in the construction of constitutive gene expression cassettes, inducible gene expression systems, and specific protein targeting systems for these bacteria. These developments are important in the food industry where lactic acid bacteria can be exploited as food-grade cell factories. PMID- 10383868 TI - DGGE/TGGE a method for identifying genes from natural ecosystems. AB - Five years after the introduction of denaturing gradient gel electrophoresis(DGGE) and temperature gradient gel electrophoresis (TGGE) in environmental microbiology these techniques are now routinely used in many microbiological laboratories worldwide as molecular tools to compare the diversity of microbial communities and to monitor population dynamics. Recent advances in these techniques have demonstrated their importance in microbial ecology. PMID- 10383869 TI - Oligosaccharide recognition signals and defence reactions in marine plant-microbe interactions. AB - Recent findings on the involvement of oligosaccharide signals in pathogen recognition and defence reactions in marine algae shine a new light on the ecology of their interactions with associated microorganisms. Since the marine environment encompasses lineages that have diverged a long time ago from the terrestrial phyla, these results suggest that cell-cell recognition pathways typical of terrestrial plants appeared very early in the evolution of eukaryotes. Production of oligosaccharides from marine algae using microbial recombinant polysaccharidases is also of industrial interest as plants can be protected from infections by preincubation in the presence of appropriate signals that mimic the attacks by pathogens. PMID- 10383870 TI - Bioprospecting in the developing world. AB - During the past ten years, species-rich nations in the developing world have sought to capitalize on their 'biological patrimony' through a variety of business relationships with multinational corporations as a means of underwriting their conservation efforts. Initially lauded, these relationships have generated more rhetoric than revenues to date. The ramifications of these results on bioprospecting are discussed along with the foreseeable changes in models of collaboration. PMID- 10383871 TI - Biodesulfurization. AB - Microbial sulfur-specific transformations have been identified that selectively desulfurize organic sulfur compounds in fossil fuels. Recent discoveries related to biodesulfurization mechanisms may lead to commercial applications of biodesulfurization through engineering recombinant strains for over-expression of biodesulfurization genes, removal of end product repression, and/or by combining relevant industrial and environmental traits with improvements in bioprocess design. PMID- 10383872 TI - Copper associated liver diseases in children. Introduction. PMID- 10383873 TI - An overview of the metabolism of copper. AB - Studies from animal models and in human volunteers have permitted to construct a provisional continuum of acceptable intakes of copper that would avoid copper deficiency and/or toxicity: acceptable intakes may vary between 10 and 50 mg/kg body weight. PMID- 10383874 TI - Diarrhoea following contamination of drinking water with copper. AB - Three cases of children with suspected copper intoxication from the drinking water are described. The children presented with protracted diarrhoea, which promptly disappeared, when they were given drinking water of low copper concentration but reappeared when given their domestic water. It is concluded that the use of copper tubing in the water pipes may under certain circumstances result in the presence of copper in the drinking water and the risk of intoxication, especially in small children. PMID- 10383875 TI - Chronic poisoning by copper in tap water: I. Copper intoxications with predominantly gastointestinal symptoms. AB - Copper can induce acute and chronic intoxications in humans. Copper in tap water has caused a series of severe systemic diseases in Germany in recent years (copper induced liver cirrhosis). Besides cirrhosis, another type of disease with predominantly gastrointestinal symptoms has occurred which likewise appeared to be induced by copper in tap water. - In a retrospective investigation we looked for additional indications and proof that chronic copper poisoning has been the cause of the observed gastrointestinal diseases. All patients suffering from this type of disease had copper plumbing in their houses. - The patients (children and adults) suffered from nausea, vomiting, colic, and diarrhoea. In the group of infants, one refused formula milk (prepared with tap water) and the others suffered from persistent restlessness, unexplainable screaming (especially at night) and/or long lasting diaper rash. - We accept the diagnosis of chronic copper intoxication as the cause of the gastrointestinal symptoms when at least one of the following criteria were fulfilled: 1. first manifestation, remission and relapse of the disease depend on intake and a non-intake of water containing copper, respectively. 2. hypercupric state of the patients (i.e. pathological high concentrations of the non-ceruloplasmin-bound copper in serum and/or elevated copper levels in urine) 3. signs of systemic copper intoxication in the same patient 4. signs of systemic copper intoxication or hypercupric states in members of the patient s family or in his neighbourhood (non-relatives) - We found that the disease can even be caused by copper concentrations below the allowed concentration given by the German Guidelines for Drinking Water (Trinkwasserverordnung). - The data prove that copper in drinking water can cause gastrointestinal diseases and not only the better known systemic diseases (i.e. copper induced liver cirrhosis). Copper poisoning must be considered as a possible cause of chronic gastrointestinal diseases in those countries in which copper plumbing is common. PMID- 10383877 TI - Pathomorphology of the liver in exogenic infantile copper intoxication in Germany. AB - Pathomorphology of the liver has been reviewed in twelve German infants with chronic exogenic copper intoxication. Eight cases suffered from florid Indian childhood cirrhosis. Seven of these children died because of liver failure. One child received liver transplantation. In contrast, four children with a stable clinical course had a complete micronodular cirrhosis with only slight developed features of liver cell damage and stable clinical course. The copper content varied between 541 microg/g dry weight (norm <59 microg/g) and 698 microg/g fresh weight (norm <5 microg/g). There was no significant difference in the copper content between the children with florid Indian childhood cirrhosis and those with micronodular cirrhosis. The results show that there exists a spectrum of pathomorphological alterations in exogenic infantile copper disease correlating with the clinical outcome in contrast to the copper content of the liver. Copper intoxication of the liver should be of diagnostic concern in any unclear case of micronodular cirrhosis in early infancy. PMID- 10383876 TI - Chronic poisoning by copper in tap water: II. Copper intoxications with predominantly systemic symptoms. AB - Copper can induce acute and chronic intoxications in humans. Copper in tap water has caused a series of severe systemic diseases in Germany in recent years (chronic copper poisoning, CCuP). From the clinical point of view it has been difficult to establish the diagnosis on the basis of clinical and laboratory methods. In a retrospective study, we therefore looked for essential clinical signs as well as laboratory findings which might be typical and essential for the diagnosis of CCuP. - We observed that in patients with severe systemic CCuP not only the liver but also several other organs have been the target of copper. As a proof copper overload has been measured. The latter results are presented here. - During or shortly after exposure "free" serum copper (= non-ceruloplasmin-bound copper) was significantly elevated in all patients (range 5.1 to 47.1 micromol/l, or 25.7 to 56.2 % of total serum copper). The normal upper limits in infants according to Salmenpera (8) are: 0.3 micromol/l, or 1.6 % of total serum copper. Total serum copper was elevated in 14/16 patients: 13.7 to 30.1 micromol/l in sick infants (normal upper level: 12.6 micromol/l), and 17.0 to 27.2 in sick children (normal upper level for children and adults: 21.4 micromol/l). - Urine copper excretion was found elevated in 9/10 patients, with a range of 11 to 456 microg/dl (normal upper level in adults: 15 microg/dl). - Our results show that patients with systemic CCuP are in a "hypercupric" state. The data thus firstly prove that indeed the putative agent copper is found in excess in the patients and secondly show that the estimation of "free" copper in serum and the measurement of copper in urine are reliable diagnostic methods. Elevation of total serum copper (even though not specific) can give a first hint to the diagnosis. - The hypercupric state of systemic CCuP can be differentiated from that of Wilson's disease by (1) normal levels of ceruloplasmin and (2) the observation that values for free copper in serum or urinary copper normalize in an environment without copper in tap water, for instance in a hospital. PMID- 10383878 TI - Early childhood cirrhoses (ECC) in Germany between 1982 and 1994 with special consideration of copper etiology. AB - In a multicentric retrospective clinical study with 16 pediatric centres we identified 103 cases of histologically confirmed early childhood cirrhosis (ECC) in Germany for the years 1984-1994. The most prominent diagnoses were congenital bile duct anomalies (47.5%), inborn metabolic disorders (17.5%) and unclear etiologies (17.5%). Chronic and excessive intake of copper might be discussed as an etiological factor in 8 other cases. 5 of these were proven to have coincided with very high hepatic copper contents and copper plumbing/acid well water. Their connection with copper exposure must be considered as probable, whereas 3 others were only suspected copper cases, mainly due to reliable exclusion of other etiologies. High corrosivity (base capacity) values and copper levels in the water for infants formula of 9-26.4 mg/L were determined in those probable cases for which exposure conditions could be exactly reproduced. Additional reports on copper associated ECC, either Indian Childhood Cirrhosis (ICC) from outside India or so-called Idiopathic Copper Toxicosis ( ICT ), originate from Austria, Australia, Germany, Ireland, USA. PMID- 10383879 TI - Factors to be considered concerning the corrosion of copper tubes. AB - Various corrosion processes in copper tubes may lead to elevated copper concentrations in drinking water, if the water is stagnant. Incorrect application of copper tubes in households may lead to elevated copper concentrations in tap water. Owners of private wells should carefully check the properties of the water to be used for human consumption. PMID- 10383880 TI - Copper and the liver: late lessons from the Camperdown experience. AB - In 1973 copper-associated liver disease was first described in an Australian boy of non-Indian origin who died at the age of 15 months with liver cirrhosis. At necropsy the hepatic copper concentration was excessively high (> 300 mg/g dry weight). It was shown that the boy s disease was caused by consumption of copper contaminated drinking water. PMID- 10383881 TI - Copper toxicosis in an Australian child. AB - A 20 month old Caucasian child living in rural Australia presented with liver failure after exposure from birth to milk formula made from acidic bore water containing excess copper leached from the copper piping. The liver pathology was identical to that seen in Indian childhood cirrhosis and similar disease in non Indian children now termed idiopathic copper toxicosis (ICT). The only other Australian case, reported more than 20 years previously, had identical presentation, pathology and circumstances of occurrence. The rarity of ICT in Australia, despite a significant population at risk, and the implications of these cases are discussed. PMID- 10383882 TI - A case of adult chronic copper self-intoxication resulting in cirrhosis. AB - Copper "supplements" taken in a dose of 30-60 mg/day during 3 years caused severe liver cirrhosis necessitating orthotopic liver transplantation. PMID- 10383883 TI - Treatment of copper associated liver disease in childhood. AB - BACKGROUND/AIMS: Copper associated liver disease is accompanied with high liver copper concentrations and progressive liver disease in infancy or childhood. The disease is thought to be due to excessive dietary copper overload (copper enriched water supply) and in addition to be based on a genetic predisposition. Treatment with penicillamine in Indian childhood disease, which probable has the same etiology, is effective when it is started early enough as well as in Wilson's disease. We aimed to describe the clinical features of copper associated liver disease and report our experience with different treatment options in German children. METHODS/RESULTS: Two boys presented at the age of 6 and 10 months with abdominal distension due to hepatosplenomegaly. The diagnosis of copper associated liver disease was made based on feeding history, standard liver function parameters, liver biopsy and assessment of dry weight copper concentration and urinary excretion of copper. Both had micronodular cirrhosis, ballooning of hepatocytes and Mallory bodies. In child A improvement of liver function was observed after introduction of penicillamine therapy and copper reduced diet. The treatment was stopped after 18 months, when normalisation of copper concentration in the liver had occured. In child B acute liver failure developed despite initiation of treatment. The boy was transplanted successfully. Both children are presently healthy 10 years after transplantation and 4 years after begin of chelating therapy, respectively. CONCLUSIONS: We conclude, that early chelating therapy with penicillamine can be successful in children with copper associated liver disease. In case of delayed diagnosis and acute liver failure liver transplantation is necessary. Our case reports highlight the urgent need of rapid diagnosis of copper associated liver disease in order to initiate early chelating therapy. Copper associated liver disease should obviously be considered in liver disease of unknown origin. Possible causes of excessive dietary copper intake should be ascertained. PMID- 10383884 TI - The tamoxifen dilemma. AB - The anti-oestrogen tamoxifen is widely used for adjuvant therapy in the treatment of women with breast cancer and has a low incidence of serious side-effects. It could also play a role as a breast cancer chemopreventive agent. However, epidemiological studies in both tamoxifen-treated breast cancer patients and in healthy women have shown that treatment results in a small increase in the incidence of endometrial cancers. While the use of tamoxifen in breast cancer patients is clearly justified, the situation for its use as a chemopreventive agent in healthy women is not so clear cut. Reasons for caution come from studies in rats that show that tamoxifen is a genotoxic mutagenic liver carcinogen. Initiation of tumours in the rat is the result of metabolic activation of tamoxifen by CYP enzymes to an electrophile(s) that binds irreversibly to DNA. This is not related to the oestrogen receptor status of the tissue. The extent of DNA damage, detected by 32P-post-labelling or accelerator mass spectrometry, is dependent both on the dose and the length of exposure. Studies have been carried out to see if such binding occurs in the uterine endometrium from tamoxifen treated women. Results are presently inconclusive, but if such irreversible DNA binding occurs, it is at very low levels. Based on a mechanistic understanding of tamoxifen-induced liver carcinogenesis in the rat, it seems that in humans hepatic DNA damage will be close to the limit of detection by 32P-post-labelling and liver cancer will not be a significant carcinogenic risk. We cannot be certain of the mode of action of tamoxifen that results in the increase in endometrial cancers in treated women but it seems unlikely that this will be associated with a classical genotoxic mechanism. PMID- 10383885 TI - Taxol, vincristine or nocodazole induces lethality in G1-checkpoint-defective human astrocytoma U373MG cells by triggering hyperploid progression. AB - In this report, we describe a novel lytic mechanism exploited by antimicrotubule drugs (AMDs) such as Taxol which are frequently used to treat multiple human cancers including breast and ovarian cancers. In cells lacking the G1-arresting capacity due to the defect in retinoblastoma or p53 gene function, AMDs trigger hyperploid progression and death. The hyperploid progression occurs via continued cell-cycle progression without cell division. Blocking hyperploid progression through hydroxyurea or ectopically expressed p27(Kip1), a G1-specific Cdk inhibitor, abrogates AMD cytotoxicity. Thus, AMDs induce lethality in G1 checkpoint-defective cells by triggering hyperploid progression. The phenomenon is reminiscent of that observed previously with bub-1 yeast mutant. The potential significance of this finding lies in that hyperploid progression-mediated death may be exploited to develop a therapy with tumor-specificity at the genetic level. As a large fraction of human cancers are mutated in p53 gene, it may have a wide therapeutic applicability. PMID- 10383886 TI - Resistance to the promotion of glutathione S-transferase 7-7-positive liver lesions in Copenhagen rats. AB - Previously, we have shown that Copenhagen (Cop) rats are highly resistant to the induction of putative preneoplastic, glutathione S-transferase 7-7 (GST 7-7) positive liver lesions following treatment with a modified resistant hepatocyte protocol. The objective of the current study was to establish the time course for the development of resistance and examine potential resistance mechanisms in Cop rats using F344 rats as susceptible controls. Male Cop and F344 rats (n = 25), 7 8 weeks of age, were initiated with diethylnitrosamine (200 mg/kg) and promoted 3 weeks later with four doses of 2-acetylaminofluorene (20 mg/kg) and a 2/3 partial hepatectomy (PH). Groups of rats from each strain were killed on days 2, 4, 7, 14 and 21 post-PH, 2 h after receiving bromodeoxyuridine. Cop livers contained similar numbers of GST 7-7-positive lesions to F344 livers on days 2 and 4 post PH. The percent volume of liver occupied by these lesions did not differ between the strains on days 2, 4 and 7 post-PH. On day 14, however, approximately 29% of the liver volume in F344 rats was occupied by lesions, whereas in Cop rats this was significantly less (approximately 9%, P < 0.001). On day 21, lesions occupied approximately 58% of F344 rat livers and only approximately 6% of Cop livers. Despite these differences, the labeling index of hepatocytes was not significantly different between the strains at any time point, either within lesions or within surrounding normal liver. Furthermore, the apoptotic indices were not different between the strains at any time. However, differences were found in the extent of lesion remodeling (redifferentiation) and in the pattern of oval cell response following PH in Cop livers. By day 14 post-PH, approximately 76% of Cop liver lesions showed evidence of remodeling, compared with only approximately 14% of F344 lesions. The oval cell response to PH was equivalent in the two strains up to day 4 post-PH but by day 7, in F344 livers there was extensive migration of these cells into the liver parenchyma, whereas in Cop livers, the response remained localized to the portal regions. These results suggest that Cop resistance occurs at the promotion stage and not the initiation stage of carcinogenesis. Resistance appears not to be due to a lower proliferation rate nor to a higher apoptotic rate within Cop lesions. Precocious remodeling and/or a diminished oval cell response, however, may contribute to the resistance of Cop rats to the growth of GST 7-7-positive hepatic lesions. PMID- 10383887 TI - Nickel(II) increases the sensitivity of V79 Chinese hamster cells towards cisplatin and transplatin by interference with distinct steps of DNA repair. AB - Nickel compounds are carcinogenic to humans and to experimental animals. In contrast to their weak mutagenicity, they have been shown previously to increase UV-induced cytotoxicity and mutagenicity and to interfere with the repair of UV induced DNA lesions by disrupting DNA-protein interactions involved in DNA damage recognition. In the present study we applied cisplatin, transplatin and mitomycin C to investigate whether these enhancing effects and DNA repair inhibition are also relevant for other DNA damaging agents. Nickel(II) at non-cytotoxic concentrations of 50 microM and higher caused a pronounced increase in cisplatin , transplatin- and mitomycin C-induced cytotoxicity, which was neither due to an altered uptake of cisplatin or transplatin nor to an increase in DNA adduct formation. However, nickel(II) inhibited the repair of cisplatin- and transplatin induced DNA lesions. In combination with transplatin, it decreased the incision frequency, indicating that the DNA damage recognition/incision step during nucleotide excision repair is affected in general by nickel(II). In support of this, concentrations as low as 10 microM nickel(II) decreased binding of the xeroderma pigmentosum complementation group A protein to a cisplatin-damaged oligonucleotide. When combined with cisplatin, the incision frequency was affected only marginally, while nickel(II) led to a marked accumulation of DNA strand breaks, indicating an inhibition of the polymerization/ligation step of the repair process. This effect may be explained by interference with the repair of DNA-DNA interstrand crosslinks induced by cisplatin. Our results suggest that nickel(II) at non-cytotoxic concentrations inhibits nucleotide excision repair and possibly crosslink repair by interference with distinct steps of the respective repair pathways. PMID- 10383888 TI - A human prostatic stromal myofibroblast cell line WPMY-1: a model for stromal epithelial interactions in prostatic neoplasia. AB - Here we report the characterization of an SV40 large-T antigen-immortalized stromal cell line, WPMY-1, derived from the same prostate as our previously described epithelial cell lines. The WPMY-1 cells were determined to be myofibroblasts on the basis of co-expression of smooth muscle alpha-actin and vimentin. They also show positive staining for androgen receptor, large-T antigen, and positive but heterogeneous staining for p53 and pRb. Their growth is stimulated by the synthetic androgen mibolerone to 145% of control (100%). Platelet-derived growth factor BB, epidermal growth factor and basic fibroblast growth factor, at 10 ng/ml, stimulated growth to 138, 143 and 146% of control, respectively. Transforming growth factor-beta, at 10 ng/ml, inhibited serum induced growth to 65% of control in the presence of 1% serum, and bFGF-induced growth to 30% of control. A serum-free medium was developed for optimal growth of WPMY-1 cells. They show anchorage-independent growth in soft agar. Studies on paracrine interactions show that myofibroblast-conditioned medium causes a marked inhibition of growth in WPE1-10 cells, while conditioned medium from WPE1-10 prostatic epithelial cells caused only a small increase in the growth of WPMY-1 cells. WPMY-1 cells secrete very low levels of MMP-9 but high levels of MMP-2, markedly higher than the epithelial cells. These epithelial and myofibroblast cell lines, derived from the same prostate, provide novel and useful models for studies on paracrine stromal-epithelial interactions in carcinogenesis, tumor progression, prevention and treatment of prostate cancer and benign prostatic hyperplasia. PMID- 10383889 TI - Genetic toxicity of cocaine. AB - Cocaine is a widely abused drug. Recently, it has been shown to induce teratogenesis in both humans and animals. Cocaine-induced teratogenicity has been associated with reactive oxygen species (ROS), which are generated by cytochrome P450 during cocaine biotransformation. Since ROS have been reported to induce genotoxicity, it is of interest to know whether cocaine and/or its metabolites are also genotoxic. In this study, Chinese hamster ovary K1 cells were employed as a model system to investigate the genetic toxicity of cocaine in the presence or absence of rat liver S9 fraction. Cocaine-induced cytotoxicity was potentiated when S9 was present, indicating the cytochrome P450 metabolism plays a role in cocaine-mediated cytotoxicity. Cocaine treatments per se induced a few chromosome aberrations while treatments of cocaine plus S9 caused a significant increase in chromosome aberrations. In contrast, cocaine induced micronuclei (MN) formation and hypoxanthine-guanine phosphoribosyltransferase mutation only in the presence of S9. Therefore, cocaine itself is at best a weak clastogen, whereas metabolite(s) of cocaine is/are truly inducer(s) of clastogenesis and mutagenesis. Cocaine treatments alone also induced a significant increase in sister chromatid exchange frequency but the addition of S9 did not affect the results. Free radical scavengers, including superoxide dismutase and catalase, efficiently decreased the frequency of cocaine plus S9-induced MN, implying that ROS are indeed important components in cocaine-induced genotoxicity. The observation that non-toxic doses of cocaine can inhibit intercellular metabolic cooperation suggests that cocaine may also be a tumor promoter. Our data supports that cocaine could possess genotoxicity in addition to its well-known neurotoxicity and teratogenicity. PMID- 10383890 TI - Altered MAP kinase (ERK1,2) regulation in primary cultures of mammary tumor cells: elevated basal activity and sustained response to EGF. AB - An elevation in total MAP kinase activity and expression has been observed in breast cancer tissue. However, the mechanisms underlying these changes in kinase activity and regulation by growth factors are not well characterized. In these studies, the effect of the potent mammary mitogen, epidermal growth factor (EGF), on the activation of the mitogen-activated protein kinases, ERK1 and ERK2 (extracellular regulated protein kinases 1 and 2, respectively), was compared in primary cultures of normal mouse mammary epithelial cells and in a hormone responsive mouse mammary tumor. In normal epithelium, EGF stimulated an early rise in ERK activity at 4 min followed by a rapid decline, whereas a sustained (1 h) elevation of ERK activity was observed in the tumor cells. The time course of ERK activity in both cell types coincided with the phosphorylation state of the EGF receptor, suggesting that altered regulation of EGF receptor phosphorylation or EGF receptor turnover produces an enhanced ERK response to EGF in tumor cells. The MEK inhibitor, PD 098059 inhibited EGF-stimulated proliferation and ERK activity in a parallel, dose-dependent manner showing that ERK activation is at least permissive for the proliferative response to EGF. In addition, tumor cells showed a 4-fold elevation in basal (or ligand-independent) activity over normal cells without an increase in total enzyme level, and a preferential activation of ERK1 by EGF. These EGF-dependent and -independent changes in ERK regulation in the hormone-responsive mammary tumor underscore how multiple alterations in the regulation of this pathway may play a role in mammary tumorigenesis. PMID- 10383891 TI - G1-arrested FaO cells re-enter the cell cycle upon stimulation with the rodent non-genotoxic hepatocarcinogen nafenopin. AB - The peroxisome proliferators are rodent non-genotoxic hepatocarcinogens that suppress apoptosis and induce DNA replication, cell proliferation and liver tumours. In order to investigate the effect of peroxisome proliferators on cell cycle progression, we arrested the well-differentiated rat hepatoma cell line FaO in the G1 phase of the cell cycle. Under these conditions, CDK2 and CDK4 protein expression remained unchanged compared with proliferating cells, but expression of cyclin D1 and p27(KIP1) was down-regulated and cyclin E accumulated in the inactive form. G1-arrested cells were able to enter the cell cycle on addition of exogenous growth factors such as epidermal growth factor (EGF) or hepatocyte growth factor (HGF) and replicate their DNA within 12 to 24 h of re-stimulation. Upon release from G1 arrest, CDK2 protein expression was down-regulated and, surprisingly, p27(KIP1) expression was restored. Cyclin D1 and phosphorylated cyclin E accumulated at 12 h but were degraded by 24 h after addition of EGF. Importantly, the peroxisome proliferator nafenopin and tumour necrosis factor alpha were able to induce DNA replication. Thus, the profile of expression of cell cycle regulatory proteins upon stimulation with nafenopin is comparable with that induced by growth factors such as EGF. PMID- 10383892 TI - cDNA cloning, expression and activity of a second human aflatoxin B1-metabolizing member of the aldo-keto reductase superfamily, AKR7A3. AB - The aflatoxin B1 (AFB1) aldehyde metabolite of AFB1 may contribute to the cytotoxicity of this hepatocarcinogen via protein adduction. Aflatoxin B1 aldehyde reductases, specifically the NADPH-dependent aldo-keto reductases of rat (AKR7A1) and human (AKR7A2), are known to metabolize the AFB1 dihydrodiol by forming AFB1 dialcohol. Using a rat AKR7A1 cDNA, we isolated and characterized a distinct aldo-keto reductase (AKR7A3) from an adult human liver cDNA library. The deduced amino acid sequence of AKR7A3 shares 80 and 88% identity with rat AKR7A1 and human AKR7A2, respectively. Recombinant rat AKR7A1 and human AKR7A3 were expressed and purified from Escherichia coli as hexa-histidine tagged fusion proteins. These proteins catalyzed the reduction of several model carbonyl containing substrates. The NADPH-dependent formation of AFB1 dialcohol by recombinant human AKR7A3 was confirmed by liquid chromatography coupled to electrospray ionization mass spectrometry. Rabbit polyclonal antibodies produced using recombinant rat AKR7A1 protein were shown to detect nanogram amounts of rat and human AKR7A protein. The amount of AKR7A-related protein in hepatic cytosols of 1, 2-dithiole-3-thione-treated rats was 18-fold greater than in cytosols from untreated animals. These antibodies detected AKR7A-related protein in normal human liver samples ranging from 0.3 to 0.8 microg/mg cytosolic protein. Northern blot analysis showed varying levels of expression of AKR7A RNA in human liver and in several extrahepatic tissues, with relatively high levels in the stomach, pancreas, kidney and liver. Based on the kinetic parameters determined using recombinant human AKR7A3 and AFB1 dihydrodiol at pH 7.4, the catalytic efficiency of this reaction (k2/K, per M/s) equals or exceeds those reported for other enzymes, for example cytochrome P450s and glutathione S-transferases, known to metabolize AFB1 in vivo. These findings indicate that, depending on the extent of AFB1 dihydrodiol formation, AKR7A may contribute to the protection against AFB1 induced hepatotoxicity. PMID- 10383893 TI - Genetic polymorphism of CYP2D6, GSTM1 and NAT2 and susceptibility to haematological neoplasias. AB - Xenobiotic-metabolizing enzymes constitute an important line of defence against a variety of carcinogens. Many are polymorphic, constituting the basis for the wide inter-individual variation in metabolic capacity and possibly a source of variation in the susceptibility to chemical-induced carcinogenesis. The aim of this study was to determine the existence of any association between the main genetic polymorphisms of cytochrome P450 2D6 (CYP2D6), glutathione S-transferase M1 (GSTM1) and N-acetyltransferase 2 (NAT2) and an altered risk for haematological neoplasias. A total of 160 patients and 128 controls were genotyped by means of PCR-RFLP-based assays. Mutated alleles comprising CYP2D6*4, GSTM1*0, NAT2*5A, *5B, *5C, *6 and *7 were analysed along with the wild-type alleles. The results showed a higher frequency of CYP2D6 extensive metabolizers carrying two functional alleles in the leukaemia group, when compared with controls (76.6 versus 57.0%, P = 0.008). No differences were found in the case of Hodgkin and non-Hodgkin lymphomas. Analysis of the GSTM1 and NAT2 polymorphisms failed to show an association with any of the neoplasias, although a near significant increase in fast acetylators was also found in the leukaemia group (50.0 versus 35.9%, P = 0.06). The results suggest an association of extensive metabolism with an increased risk for leukaemia, possibly by an increase in the metabolic activation of chemical carcinogens or linkage to another cancer-causing gene. Opposite findings presented in other studies may reflect geographical differences in the type of environmental carcinogens to which different populations are exposed. PMID- 10383894 TI - Interaction between haemochromatosis and transferrin receptor genes in different neoplastic disorders. AB - A number of genes are involved in iron metabolism, including the transferrin receptor (TFR) and haemochromatosis (HFE) genes. In previous investigations an increased risk for neoplastic disease has been observed in individuals homo- and heterozygous for hereditary haemochromatosis. The HFE wild-type gene product complexes with the transferrin receptor (TF) and two different HFE mutations (Cys282Tyr and His63Asp) have been found to increase the affinity of TFR for TF and increase cellular iron uptake. In a recent study we found no associations for HFE and TFR separately, but an interaction between HFE and TFR genotypes in multiple myeloma. Individuals carrying the HFE Tyr282 allele (homo- and heterozygotes) in combination with homozygosity for the TFR Ser142 allele had an increased risk. In the present study the same association was found in breast and colorectal cancer. The odds ratio for all three neoplasms combined was 2.0 (95% CI 1.0-3.8). The risk for neoplastic disease was further increased (OR 7.7, 95% CI = 1.0-59.9) when the analysis was restricted to HFE Tyr homozygotes and compound heterozygotes in combination with TFR Ser homozygosity. Thus, an interaction between HFE and TFR alleles may increase the risk for different neoplastic disorders. PMID- 10383896 TI - 32P-postlabeling high-performance liquid chromatography (32P-HPLC) adapted for analysis of 8-hydroxy-2'-deoxyguanosine. AB - 8-Hydroxy-2'-deoxyguanosine (8-OH-dG) is a promutagenic lesion in DNA caused by reactive oxygen species. It normally exists at a level of 0.1-1 per 10(5) 2' deoxyguanosines (dG). To analyze the lesion in easily obtainable biological samples, a very sensitive analytical method is required. The method should also handle the problem with potential oxidation of dG to 8-OH-dG during workup and analysis. 32P-postlabeling high-performance liquid chromatography (32P-HPLC) is an analytical method previously used to analyze lipophilic DNA adducts at levels as low as 1 per 10(9) normal nucleotides when analyzing microgram amounts of DNA. This method was adapted for analysis of 8-OH-dG. The aim was to develop an analytical method that provided a high sensitivity and good reproducibility, prevented oxidation of dG present in samples to 8-OH-dG, was capable of analyzing DNA from very small samples and still offered high sample throughput and ease of use. In analysis of calf thymus DNA, the method had a detection limit of 0.1 8-OH dG per 10(5) dG when 1 microgram of DNA was used. The standard deviation of repeated analyses of the same sample was +/-10% and the result corresponded well with the established analytical method using HPLC with electrochemical detection. 32P-HPLC is sensitive enough to enable analysis of low levels of 8-OH-dG in biological samples such as small volumes of blood, needle biopsies and tissue swabs. It also substantially reduces oxidation of dG to 8-OH-dG during sample workup and analysis. PMID- 10383895 TI - Association of NAD(P)H:quinone oxidoreductase (NQO1) null with numbers of basal cell carcinomas: use of a multivariate model to rank the relative importance of this polymorphism and those at other relevant loci. AB - Glutathione S-transferase GSTM1 B and GSTT1 null, and cytochrome P450 CYP2D6 EM have been associated with cutaneous basal cell carcinoma (BCC) numbers, although their quantitative effects show that predisposition to many BCC is determined by an unknown number of further loci. We speculate that other loci that determine response to oxidative stress, such as NAD(H):quinone oxidoreductase (NQO1) are candidates. Accordingly, we assessed the association between NQO1 null and BCC numbers primarily to rank NQO1 null in a model that included genotypes already associated with BCC numbers. We found that only 14 out of 457 cases (3.1%) were NQO1 null. This frequency did not increase in cases with characteristics linked with BCC numbers including gender, skin type, a truncal lesion or more than one new BCC at any presentation (MPP). However, the mean number of BCC in NQO1*0 homozygotes was greater than in wild-type allele homozygotes and heterozygotes, although the difference was not quite significant (P = 0.06). These data reflect the link between NQO1 null and BCC numbers in the 42 MPP cases rather than the whole case group. We identified an interaction between NQO1 null and GSTT1 null that was associated with more BCC (P = 0.04), although only four cases had this combination. The relative influence of NQO1 null was studied in a multivariate model that included: (i) 241 patients in whom GSTM1 B, GSTT1 null and CYP2D6 EM genotype data were available, and (ii) 101 patients in whom these genotypes, as well as data on GSTM3, CYP1A1 and melanocyte-stimulating hormone receptor (MC1R) genotypes were available. NQO1 null (P = 0.001) and MC1R asp294/asp294 (P = 0.03) were linked with BCC numbers, and the association with CYP2D6 EM approached significance (P = 0.08). In a stepwise regression model only these genotypes were significantly associated with BCC numbers with NQO1 null being the most powerful predictor. PMID- 10383897 TI - Elevation of urinary enzyme levels in rat bladder carcinogenesis. AB - Urinary enzyme levels were investigated in rats administered different promoters in their diet for 32 weeks after being initiated by treatment with 0.05% N-butyl N-(4-hydroxybutyl)nitrosamine in their drinking water for 4 weeks. All groups were composed of 10 rats each. Group 1: females treated with 3% uracil (100% carcinoma incidence). Group 2: control females kept on basal diet only (0% carcinoma incidence). Group 3: males treated with 5% sodium L-ascorbate (100% carcinoma incidence). Group 4: control males (0% carcinoma incidence). Urine was collected at the end of weeks 12, 24 and 36 and tested for lactate dehydrogenase (LDH), alkaline phosphatase, N-acetyl-beta-D-glucosaminase and aspartate aminotransferase activity. To facilitate comparison, data were related to the corresponding excreted creatinine levels. All measurements were made using a centrifugal automatic analyzer. The urine of rats with cancer lesions (groups 1 and 3) showed significant elevation in all enzyme activities at weeks 24 and/or 36 except for LDH in females (group 1). The M/H ratio of the LDH isozymes was reversed (1.10 +/- 0.10) in the tested rats with carcinomas at week 36. This study thus provides evidence of a correlation between high urinary enzyme levels and cancer development in the rat bladder. Measurement of the tested enzymes might thus provide a method to detect malignant changes in bladder epithelium by direct urine analysis. PMID- 10383898 TI - Butadiene diolepoxide- and diepoxybutane-derived DNA adducts at N7-guanine: a high occurrence of diolepoxide-derived adducts in mouse lung after 1,3-butadiene exposure. AB - Butadiene (BD) is a high production volume chemical and is known to be tumorigenic in rodents. BD is metabolized to butadiene monoepoxide (BMO), diepoxybutane (DEB) and butadiene diolepoxide (BDE). These epoxides are genotoxic and alkylate DNA both in vitro and in vivo, mainly at the N7 position of guanine. In this study, a 32P-post-labeling/thin-layer chromatography (TLC)/high-pressure liquid chromatography (HPLC) assay for BDE and DEB adducts at the N7 of guanine was developed and was used in determining the enantiomeric composition of the adducts and the organ dose of BD exposure in lung. Exposure of 2'-deoxyguanosine (dGuo), 2'-deoxyguanosine-5'-phosphate (5'-dGMP) and 2'-deoxyguanosine-3' phosphate (3'-dGMP) to racemic BDE followed by neutral thermal hydrolysis gave two products (products 1 and 2) that were identified by MS and UV and NMR spectroscopy as a diastereomeric pair of N7-(2,3,4-trihydroxybutan-1-yl) guanines. Exposure of dGuo nucleotides to RR/SS DEB (also referred to as dl DEB) followed by thermal depurination resulted in a single product coeluting with the BDE product 1. If the reaction mixture of BDE and 5'-dGMP was analyzed by HPLC before hydrolysis of the glycosidic bond, four major nucleotide alkylation products (A, B, C and D) with identical UV sepectra were detected. The products were isolated and hydrolyzed, after which A and C coeluted with product 1 and B and D coeluted with the product 2. The major adduct of DEB-exposed 5'-dGMP was N7 (2-hydroxy-3,4-epoxy-1-yl)-dGMP (product E). A 32P-post-labeling assay was used to detect BDE- and DEB-derived N7-dGMP adducts in DNA. Levels of adducts increased with a dose of BDE and DEB and exhibited a half life of 30 +/- 3 (r = 0.98) and 31 +/- 4 h (r = 0.95), respectively. Incubation of DEB-modified DNA at 37 degrees C at neutral pH for up to 142 h did not lead to an increase of N7 (2,3,4-trihydroxybutan-1-yl)-dGMP in the DNA. These observations led to the conclusion that the N7-(2,3, 4-trihydroxybutan-1-yl)-dGMP adducts in DNA can be used as a marker of BDE exposure and that N7-(2-hydroxy-3,4-epoxy-1-yl)-dGMP adducts are related to DEB exposure. Dose-related levels of BDE- and DEB-derived adducts were detected in lungs of mice inhaling butadiene. Most of the N7-dGMP adducts (73%; product D) were derived from the 2R-diol-3S-epoxide of 1,3 butadiene. The data presented in this paper indicate that in vivo, 98% of N7-dGMP alkylation after BD exposure is derived from BDE, and approximately 2% of the adducts were derived from DEB and BMO. PMID- 10383899 TI - Effect of Helicobacter pylori infection on the N-methyl-N'-nitro-N nitrosoguanidine-induced gastric carcinogenesis in mongolian gerbils. AB - The effect of Helicobacter pylori infection on N-methyl-N'-nitro-N nitrosoguanidine (MNNG)-induced gastric cancer was studied using a Mongolian gerbil model. Five-week-old male Mongolian gerbils were divided into four groups of 25-30 animals each and challenged for 20 weeks with H.pylori, MNNG, a combination of H.pylori and MNNG, or neither of them. Four to 20 animals from each group were killed at 16, 24 and 52 weeks after H.pylori inoculation, and histopathological changes in their stomachs were examined. A well-differentiated adenocarcinoma was first observed 24 weeks after inoculation in the combination group. At 52 weeks, only six of 15 animals were colonized with H.pylori persistently, and four of them showed well-differentiated adenocarcinomas; on the other hand, neither of the animals with disappearance of H.pylori from the combination group showed adenocarcinoma. At the same observation time, three of 17 animals from MNNG group showed poorly differentiated adenocarcinomas. The incidence of gastric carcinoma in the combination group was significantly higher than that in the MNNG group (P < 0.05). However, no tumors were seen in the control and H.pylori groups. The present findings demonstrate that H.pylori infection enhances the carcinogenic action of MNNG. PMID- 10383900 TI - In vivo effects of ascorbate and glutathione on the uptake of chromium, formation of chromium(V), chromium-DNA binding and 8-hydroxy-2'-deoxyguanosine in liver and kidney of osteogenic disorder shionogi rats following treatment with chromium(VI). AB - Several previous in vitro studies have indicated that ascorbate and glutathione are the major reductants of Cr(VI) in cells. In order to evaluate the in vivo effects of ascorbate and glutathione on Cr(VI)-induced carcinogenesis, Cr uptake and the formation of Cr(V), Cr-DNA adducts and 8-hydroxy-2'-deoxyguanosine (8-OH dG) were measured in the liver and kidney of Osteogenic Disorder Shionogi (ODS) rats that lack the ability to synthesize ascorbate. Despite a 10-fold difference in tissue ascorbate levels among different dietary ascorbate groups, the Cr(V) signal intensity, Cr uptake and total Cr-DNA binding were not affected in either organ. Treatment of ODS rats with Cr(VI) (10 mg/kg) had no substantial effect on the levels of ascorbate and glutathione in these tissues. The levels of Cr(V) and Cr-DNA binding were approximately 2-fold higher in the liver than in the kidney, although the levels of total Cr uptake were similar in both tissues. Cr uptake levels were significantly lower in the liver and kidney of ODS rats treated with high levels of ascorbate and a high dose of Cr(VI) (40 mg/kg), suggesting a detoxifying role played by plasma ascorbate. Similarly, modulation of glutathione levels by N-acetyl-L-cysteine, L-buthionine-S, R-sulfoximine or phorone in these animals by up to 2-fold had little or no consistent effect on Cr uptake, Cr-DNA binding, Cr(V) levels or 8-OH-dG formation in either organ. One possible explanation is that reduction of ascorbate and glutathione concentration to <10 and 50%, respectively, of normal in these two organs still provides threshold levels of these two reductants that are in excess of what is needed for significant reductive activation of Cr(VI). Alternatively, it is possible that ascorbate and glutathione do not play a major role in the formation of Cr(V), Cr DNA binding or 8-OH-dG and that other cellular reductants, such as cysteine or other amino acids, might be more important reductants of Cr(VI) in vivo. PMID- 10383901 TI - Neonatal exposure to the food mutagen 2-amino-1-methyl-6-phenylimidazo[4,5 b]pyridine via breast milk or directly induces intestinal tumors in multiple intestinal neoplasia mice. AB - We examined whether the food mutagen 2-amino-1-methyl-6-phenylimidazo[4,5 b]pyridine (PhIP) could increase intestinal tumorigenesis in neonatal C57BL/6J Min/+ mice, a murine model for familial adenomatous polyposis. Min/+ mice are heterozygous for a nonsense mutation in the adenomatous polyposis coli gene and spontaneously develop multiple intestinal adenomas, primarily in the small intestine. Neonatal Min/+ mice (3-6 days old) were exposed to PhIP via breast milk from lactating dams given 8 s.c. injections of 50 mg/kg PhIP three times a week or to 8 s.c. injections of 25 or 50 mg/kg PhIP directly, over the same period. At the age of 11 weeks, the number, diameter and location of the intestinal tumors were scored. Remarkably, a 2- to 4-fold increase in the number of small intestinal tumors was seen in Min/+ mice exposed to PhIP via breast milk (P < 0.001). To our knowledge, this is the first time PhIP has been reported to induce tumors following exposure via breast milk from PhIP-exposed dams. Upon direct exposure to 50 mg/kg PhIP, a 6- to 9-fold increase in the number of small intestinal tumors was observed (P < 0.001). The diameter of the PhIP-induced small intestinal tumors was slightly increased (P < 0.001). In the colon, a 3- to 4-fold increase in the number of tumors was seen in Min/+ mice exposed to PhIP via breast milk (P = 0. 004). Direct exposure to 50 mg/kg PhIP caused a 2- to 6 fold increase in the number of colonic tumors (P = 0.014). The PhIP-induced colonic tumors were located more distally and displayed a smaller diameter than the tumors from the controls (P < 0.05). In contrast to a previous study, where PhIP showed only a moderate tumorigenic effect in adult Min/+ mice, the present study demonstrates a strong tumorigenic effect of PhIP in neonatally exposed Min/+ mice, even after exposure via breast milk from PhIP-exposed dams. PMID- 10383902 TI - p53 mutations in tumor and non-tumor tissues of thorotrast recipients: a model for cellular selection during radiation carcinogenesis in the liver. AB - Concerns over cancer development from exposure to environmental sources of densely ionizing, high linear energy transfer (LET) radiation, such as alpha particles from radon, is a current public health issue. The study of tumors attributable to high LET irradiation would greatly augment our insights into the biological mechanisms of carcinogenesis. Chronic low-dose-rate internal exposure to alpha-radiation from thorium dioxide deposits following intravascular administration of the radiographic contrast agent Thorotrast is known to markedly increase the risk of cancer development, especially that of hepatic angiosarcomas and cholangiocarcinomas. Although the mechanism is hypothesized to be via cellular damage, DNA being a major target, wrought by the high LET alpha particles, the specific genes and the actual sequence of events involved in the process of transforming a normal cell into a malignant one are largely unknown. To shed some light on the molecular mechanisms of cancer development during a lifetime exposure to alpha-radiation, we analyzed the most commonly affected tumor suppressor gene in humans, p53, in 20 Thorotrast recipients who developed cancer, mostly of hepatic bile duct and blood vessel origin. Of the 20 cases, 19 were found to harbor p53 point mutations. Moreover, the accompanying non-tumor tissues from these patients also had p53 mutations, albeit at lower frequency. The distribution pattern of the point mutations was significantly different between the non-tumor and tumor tissues, with most mutations in malignant tissues located in the highly conserved domains of the p53 gene. Our results support the idea that p53 mutations are important in the genesis of Thorotrast-induced tumors but that these point mutations are a secondary outcome of genomic instability induced by the irradiation. Additionally, non-tumor cells harboring p53 mutations may gain some survival advantage in situ but mutations in the domains responsible for the formation of structural elements critical in binding DNA may be necessary for a cell to reach full malignancy. PMID- 10383903 TI - Sequence-dependent mutations in a shuttle vector plasmid replicated in a mismatch repair deficient human cell line. AB - We utilized a shuttle vector plasmid (pLSC) to assess the role of DNA sequence and mismatch repair on mutagenesis in human cells. pLSC contains an interrupted 29 bp mononucleotide poly(G) run within a bacterial suppressor tRNA gene, which acts as a highly sensitive mutagenic target for detection of base substitution and frameshift mutations. The frequency of spontaneous mutations in pLSC was found to be similar after replication in either the hMSH6 (GT binding protein) mismatch repair-deficient MT1 line or its parental, mismatch repair-proficient line, TK6. However, the classes of plasmid mutations showed distinct differences in the two cell lines. Single base deletions comprised 48% of the mutations in the 56 independent pLSC plasmids sequenced from MT1 cells while these represented only 18% of the 40 independent pLSC mutants sequenced from the wild-type TK6 cells (P = 0.001). Virtually all the deletions included the mononucleotide run. In contrast, in pSP189, which contains the unmodified supF tRNA without the mononucleotide sequence, no single base deletions were observed for either cell line (P < 0.001). UV treatment of pLSC and pSP189 resulted in a 12-140-fold increase in mutations in TK6 and MT1 cells. These were predominately single base substitution mutations without a large increase in deletion mutations in the mononucleotide run in pLSC. These data indicate that a mononucleotide poly(G) run promotes single base deletion mutations. This effect is enhanced in a hMSH6 mismatch repair-deficient cell line and is independent of UV-induced mutagenesis. PMID- 10383904 TI - New studies on trans-anethole oxide and trans-asarone oxide. AB - The widespread use of naturally occurring alkenylbenzenes as flavoring and fragrance agents has led to a long-standing interest in their toxicity and carcinogenicity. Among them several allyl- and propenylbenzenes have been found to be mutagenic and carcinogenic. It has been shown that the carcinogenicity of several allylbenzenes can be related to the formation of electrophilic sulfuric acid esters following 1'-hydroxylation. Unlike the allylbenzenes, the mechanisms of carcinogenesis of propenylbenzenes such as anethole and asarone are not clear. It has been reported that one of the main metabolic pathways of trans-anethole is the epoxidation of the side chain 1,2-double bond, which was responsible for cytotoxicity but not for genotoxicity. However, we report here that synthetic trans-anethole oxide prepared from trans-anethole and dimethyldioxirane is not only mutagenic for Salmonella tester strains but is also carcinogenic in the induction of hepatomas in B6C3F1 mice and skin papillomas in CD-1 mice. Synthetic trans-asarone oxide was also carcinogenic in the induction of hepatomas as well as mutagenic for Salmonella strains. Further studies are needed on these side chain oxides of trans-anethole and trans-asarone as possible metabolites in the toxicity, mutagenicity and carcinogenicity of these and other propenylbenzenes. PMID- 10383905 TI - 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) retards mammary gland involution in lactating Sprague-Dawley rats. AB - 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), a compound from cooked meat, is an established mammary gland carcinogen in female rats. Four doses of PhIP (150 mg/kg, p.o., once per day) were given to lactating Sprague-Dawley rats separated from their 10-day-old pups to initiate involution of the gland. Twenty four hours after the last dose, apoptotic index in the mammary gland, as measured by the TUNEL assay, was significantly higher in the gland from control rats than in the PhIP-treated rats (4.757 +/- 1.066 versus 1.905 +/- 0.248%; P < 0.05). In comparison with controls, alveoli in the mammary gland of PhIP-treated rats were also visibly larger and contained more secretory epithelial cells. The expression of Bax, a stimulator of apoptosis, and Bcl-2, an inhibitor of apoptosis, were quantitated by western blotting. Accordingly, Bax expression was 2.7-fold higher in control rats, whereas Bcl-2 expression was 3.1-fold higher in PhIP-treated rats, both changes being statistically different (Student's t-test, P < 0.05). Immunohistochemistry further confirmed a lower expression of Bax and higher expression of Bcl-2 in secretory alveolar epithelial cells of the PhIP-treated mammary gland. The findings are consistent with the notion that exposure to PhIP retarded involution via partial inhibition of programmed cell death. To investigate possible mechanisms for the inhibitory effects of PhIP on mammary gland involution, serum levels of prolactin, an important hormone for the maintenance of lactation, were measured in virgin rats with regular estrous cycles given PhIP (150 mg/kg, p.o.) on the morning of diestrous. After one estrous cycle, on proestrous morning, serum prolactin levels were 1.3-fold higher after PhIP than after control vehicle (one-way ANOVA, Fisher LSD multiple comparison test, P < 0. 05). PhIP exposure during involution was associated with the induction of benign mammary tumors. Seven out of 12 rats developed fibroadenomas, and one developed a tubulopapillary carcinoma within 1 year of receiving PhIP administration during involution (150 mg/kg, p.o., once per day for 5 days), and a high-fat diet (23.5% corn oil). An increase in serum prolactin level and the effects on mammary gland apoptosis seen with PhIP may have implications for the mechanisms of carcinogenic targeting of PhIP to the mammary gland. PMID- 10383906 TI - Oxazepam is mutagenic in vivo in Big Blue transgenic mice. AB - Although oxazepam (Serax), a widely used benzodiazepine anxiolytic, does not induce gene mutations in vitro or chromosomal aberrations in vivo, it was found to be a hepatocarcinogen in a 2 year bioassay in B6C3F1 mice. Thus, it was of interest to determine whether this carcinogen is mutagenic in vivo. Male B6C3F1 Big Blue transgenic mice were fed 2500 p.p.m. oxazepam or control diet alone for 180 days and killed on the next day. The mutant frequency (MF) of lacI in control mice was 5.02 +/- 2.4x10(5), whereas the MF in the oxazepam-treated mice was 9.17 +/- 4.82x10(-5), a significant increase (P < 0.05). Correction of the mutant frequency of lacI from the oxazepam-treated mice for clonality resulted in a decrease in the mean mutant frequency to 8.15 +/- 2. 54x10(-5). Although the mutant frequency difference was small, sequencing of a random collection of the mutants from each oxazepam-exposed mouse showed a significant difference (P < 0.015) in the mutation spectrum compared with that from control mice. In the oxazepam-exposed mice, an increase in G:C-->T:A and G:C-->C:G transversions and a concomitant decrease in G:C-->A:T transitions were observed. Clonal expansion of mutations at guanines in 5'-CpG-3' sequencing contexts at three sites was noted. It is postulated that some of the mutations found in the oxazepam-derived spectrum were due to oxidative damage elicited by induction of CYP2B isozymes as the result of chronic oxazepam administration. This study demonstrates that the in vivo Big Blue transgenic rodent mutation assay can detect mutations derived from a carcinogen that did not induce gene mutations in vitro or micronuclei in mouse bone marrow. Moreover, the sequencing of the recovered mutants can distinguish between the mutation spectrum from treated mice compared with that from control mice, thereby confirming the genotoxic consequences. PMID- 10383908 TI - Smoking disturbs mitochondrial respiratory chain function and enhances lipid peroxidation on human circulating lymphocytes. AB - Mitochondria constitute a source of reactive oxygen species. We tested whether mitochondrial function from human circulating lymphocytes is affected by smoking habit and if this could be associated with an increase in oxidative damage of biological membranes. We prospectively studied 35 smokers and 35 non-smoking healthy individuals matched by age and sex, with a similar physical activity. Individual enzyme activity of complexes II, III and IV of the mitochondrial respiratory chain (MRC) and of glycerol-3-phosphate dehydrogenase activity were measured spectrophotometrically. Intact cell respiration and oxidative rates after addition of pyruvate, succinate and glycerol-3-phosphate were assessed polarographically. Lipid peroxidation of biological membranes was assessed measuring the loss of cis-parinaric acid fluorescence. Results are expressed as means (+/-SD). Smokers showed a significant decrease in complex IV activity compared with non-smokers (112.8 +/- 40.9 versus 146.4 +/- 62.5 nmol/min/mg protein, respectively; 23% of inhibition; P = 0.01), while the rest of the complexes of MRC were unaffected. Conversely, oxidative rate with succinate, but not with the other substrates, was enhanced in smokers compared with non-smokers (16.7 +/- 10.4 versus 11.4 +/- 4.7 nmol oxygen/min/mg protein, respectively; 46% of activation; P = 0. 01). Lipid peroxidation of lymphocyte membranes was increased in smokers with respect to non-smokers (3.49 +/- 1.27 versus 4.39 +/- 1. 76 units of fluorescence/mg protein, respectively; 21% of increase; P = 0.03) and this increase correlated positively with succinate oxidation activation (R = 0.34, P = 0.02) and, to a lesser extent, with complex IV inhibition, although it did not reach statistical significance (R = 0.19, P = 0.18). In smokers, the MRC function of lymphocytes is disturbed and correlates with the degree of oxidative damage of membranes. This mitochondrial dysfunction could contribute to increased endogenous production of reactive oxygen species and could play a role in tobacco carcinogenicity. PMID- 10383907 TI - Expression of matrix metalloproteinase 2 (MMP-2), membrane-type 1 MMP and tissue inhibitor of metalloproteinase 2 and activation of proMMP-2 in pancreatic duct adenocarcinomas in hamsters treated with N-nitrosobis(2-oxopropyl)amine. AB - In order to assess the significance of changes in metalloproteinase activity in pancreatic carcinogenesis, the expression of matrix metalloproteinases 2 and 9 (MMP-2 and MMP-9, respectively), tissue inhibitor of metalloproteinase-1 (TIMP-1) and TIMP-2, and membrane-type 1 MMP (MT1-MMP) and MT2-MMP in ductal lesions in a rapid-production model for pancreatic duct carcinomas (PCs) in hamsters initiated with N-nitrosobis(2-oxopropyl)amine (BOP) and in subcutaneous transplantable tumors of hamster pancreatic duct carcinoma (HPDs) was investigated. Northern analysis revealed MMP-2, MMP-9, TIMP-2 and MT1-MMP mRNAs to be overexpressed in PCs. Immunohistochemically, elevated levels of MMP-2 were apparent in early duct epithelial hyperplasias and staining increased from atypical hyperplasias to carcinomas. Gelatin zymography demonstrated clear activation of proMMP-2 but not proMMP-9 in both of primary and HPD tumors, the MT1-MMP mRNA level and proMMP-2 activation being significantly correlated (r = 0.893, P < 0.001). In our rapid production model, 0.1 and 0.2% OPB-3206, an inhibitor of MMPs, given in the diet after two cycles of augmentation pressures for 48 days decreased the incidence and number of carcinomas. Gelatin zymography demonstrated that OPB-3206 inhibited activation of proMMP-2 in pancreatic cancer tissues. These results indicate that overexpression of MMP-2, TIMP-2 and MT1-MMP, and cell surface activation of proMMP-2 by MT1-MMP, are involved in the development of PCs, and that MMP-2 expression at the protein level appears in the early phase of pancreatic duct carcinogenesis. OPB-3206 may be a candidate chemopreventive agent for pancreatic ductal adenocarcinomas. PMID- 10383909 TI - Immunohistochemical localization of inducible nitric oxide synthase and 3 nitrotyrosine in rat liver tumors induced by N-nitrosodiethylamine. AB - Human liver cancers have been associated mainly with chronic inflammations such as viral hepatitis B or C. This suggests that prolonged cell damage by chronic inflammation is critical in cancer development. Overproduction of nitric oxide (NO.) and its derivative (NOx, peroxynitrite) has been implicated as a cause of tissue damage by inflammation, thus contributing to tumor promotion. We have demonstrated the expression of the inducible isoform of nitric oxide synthase (iNOS) and 3-nitrotyrosine, a marker of peroxynitrite formation, by immunohistochemistry in preneoplastic and neoplastic rat liver tissues induced by continuous infusion of N-nitrosodiethylamine with mini-pumps. The preneoplastic lesions were characterized by proliferation of phenotypically altered hepatic foci (PAHF), dysplastic hepatocytes and oval cells. Histologically, the tumors were hepatocellular carcinomas (HCCs) of trabecular, (pseudo)glandular and solid types with or without cholangiocellular involvement. iNOS was located mainly in oval cells, capillary endothelial and muscular cells, epithelia of cholangiomas and glandular HCCs. 3-Nitrotyrosine was observed in the cytoplasms of PAHF and dysplastic hepatocytes in preneoplasias and in the cytoplasms of some living or apoptotic HCC cells, connective tissues, proteinaceous fluids, sinusoidal endothelia of tumorous hepatocytes and cholangiomas in tumors. From these observations, we suggest that: (i) chronic tissue damage by chemical carcinogens may act to induce iNOS and peroxynitrite formation; (ii) oval cells play a key role in development and/or growth of tumor tissues by producing NO. via iNOS, which may also cause tissue damage by peroxynitrite; (iii) iNOS can be considered as a phenotypic marker in cells of oval cell lineage and neovascularized capillaries in tumor tissues. PMID- 10383910 TI - Detection of 1,N2-propanodeoxyguanosine adducts of 2-hexenal in organs of Fischer 344 rats by a 32P-post-labeling technique. AB - 2-Hexenal is an alpha,beta-unsaturated carbonyl compound which is mutagenic, genotoxic and forms cyclic 1,N2-propanodeoxyguanosine adducts like similar propenals for which carcinogenicity was shown, e.g. acrolein or crotonaldehyde. Since humans have a permanent intake of 2-hexenal via vegetarian food this genotoxic compound is considered to play a role in human carcinogenicity. The data base is, however, presently not sufficient for a cancer risk assessment. To date no long term carcinogenicity study on 2-hexenal has been published. Detection of respective DNA adducts of this substance in animals or humans could allow cancer risk assessment. Therefore, we have developed a 32P-post-labeling technique based on nuclease P1 enrichment and TLC separation of the labeled adducts. The respective adducts are stable over a wide pH range from pH 4 to pH 11 and relatively stable against nuclease P1. The detection limit was 0.03 adducts per 10(6) nucleotides and the recovery was 10%. With this method we have shown in vivo formation of 1,N 2-propanodeoxyguanosine adducts of 2-hexenal for the first time and found the respective DNA adducts in different organs of Fischer 344 rats after gavage of 500, 200 and 50 mg 2-hexenal/kg body wt. No adducts could be detected in the organs of untreated rats. There is a clear dependence of the adduct level and the CBI (covalent binding index) on the dose. The CBI of 2-hexenal calculated on the basis of our adduct levels is extremely low (0.06). Since intake of 2-hexenal via fruit and vegetables is very low the cancer risk from 2-hexenal intake via food must also be considered as very low according to a first raw estimation on the basis of CBI and intake. The situation deserves, however, a more precise risk assessment in the future. PMID- 10383912 TI - Effect of mitogenic or regenerative cell proliferation on lacz mutant frequency in the liver of MutaTMMice treated with 5, 9-dimethyldibenzo[c,g]carbazole. AB - The purpose of this work was to investigate the impact of cell proliferation on liver mutagenesis. The genotoxic hepatocarcinogen 5, 9 dimethyldibenzo[c,g]carbazole (DMDBC) was administered to lacZ transgenic MutaTMMice at a non-hepatotoxic dose of 10 mg/kg, which induces only a slight increase in the liver lacZ mutant frequency (MF). To determine if cell proliferation stimuli enhanced DMDBC mutagenicity, MF was analyzed in mice first receiving DMDBC 10 mg/kg, then approximately 2 weeks later, either carbon tetrachloride (CCl4, a cytotoxic agent inducing regenerative cell proliferation) or phenobarbital (PB, a mitogenic agent inducing direct hyperplasia). In preliminary studies, the extent of cell proliferation induced by CCl4, PB and DMDBC was determined in non-transgenic CD2F1 mice by means of 5-bromodeoxyuridine labeling. The labeling index was significantly increased after CCl4 and PB, while no change was detected with DMDBC. MF was then determined in MutaTMMice 28 days after initial DMDBC treatment. No increase in MF was detected in mice receiving CCl4 or PB alone. A 2- to 3-fold increase in MF was detected in mice treated with 10 mg/kg DMDBC alone. In contrast, MF was markedly increased in mice receiving DMDBC followed by proliferative treatment (15-fold with CCl4 and 25-fold with PB). These results demonstrate that expression of DMDBC-induced mutations in mouse liver largely depends on the induction of cell proliferation (by a cytotoxic or mitogenic stimulus) and illustrate that MutaTMMouse is a valuable tool to investigate the early events of liver carcinogenesis. PMID- 10383911 TI - Tamoxifen induces G:C-->T:A mutations in the cII gene in the liver of lambda/lacI transgenic rats but not at 5'-CpG-3' dinucleotide sequences as found in the lacI transgene. AB - Tamoxifen, a rat liver carcinogen, can induce mutations in the lacI gene in the livers of lambda/lacI transgenic rats. However, the presence of persistent tamoxifen adducts on the liver DNA raises the possibility that some contribution to the mutagenesis from ex vivo mutations during the in vitro lacI assay cannot be ruled out. To address this issue, mutagenesis at the cII gene of the transgenic shuttle vector was determined using a selection based assay which is unaffected by the presence of tamoxifen-DNA adducts. Female lambda/lacI transgenic rats were dosed orally with tamoxifen (20 mg/kg body wt) daily for 6 weeks, causing a 3.2-fold increase in the mutant frequency (MF) in the cII gene compared with that obtained with solvent treated animals. This was similar to the MF found previously at the lacI gene and confirms that tamoxifen is mutagenic in vivo. The major class of mutation induced by tamoxifen in the cII gene was G:C- >T:A transversions as was found previously in the lacI gene. However, in the one unreplicated study of mutations in the p53 gene of liver tumours induced by tamoxifen, no G:C-->T:A transversions were found; possible differences between mutagenesis in normal and tumour tissues are explored. The major proportion of the G:C-->T:A transversions occurred at 5'-CpG-3' dinucleotide (CpG) sites in the lacI gene, but not at such sites in the cII gene. The methylation of CpG sites greatly enhances the targeting of deoxyguanosine by carcinogens, thus this finding might be explained by differences in the methylation patterns at their respective CpG sites; however, nothing is known about the methylation status of either the lacI nor the cII gene in this transgenic rat. This study raises the important issue of which target genes (mammalian or transgenic) should be used as endpoints in mammalian mutagenesis assays. PMID- 10383913 TI - Analysis of loss of heterozygosity in neoplastic nodules induced by diethylnitrosamine in the resistant BFF1 rat strain. AB - Loss of heterozygosity (LOH) at specific chromosomal regions is a frequent event in poorly differentiated human hepatocellular carcinomas (HCCs), but rare in mouse HCCs. This behavior could depend on interspecies differences in mechanisms of hepatocarcinogenesis or in developmental stage of lesions. To verify if LOH is involved in rat hepatocarcinogenesis, we studied LOH frequency in slowly growing neoplastic nodules induced by Solt-Farber model in diethylnitrosamine-initiated BFF1 rats. We analyzed, with microsatellites, markers at 67 rat loci dispersed over all chromosomes, corresponding to regions homologous to those lost in human HCCs or containing hepatocellular susceptibility (Hcs) or resistance (Hcr) loci in rat and mouse. In agreement with previous findings with mouse HCCs, but at variance with human HCCs, no detectable LOH was found at any locus in rats, suggesting rare LOH involvement in neoplastic nodules, with low tendency to progress to full malignancy, of BFF1 rats. PMID- 10383914 TI - Gap junctional intercellular communication and connexin43 expression in human ovarian surface epithelial cells and ovarian carcinomas in vivo and in vitro. AB - Gap junctional intercellular communication (GJIC) and the expression of gap junction proteins (connexins) are frequently decreased in neoplastic cells and have been increased by cAMP and retinoids. GJIC and connexin expression were investigated in early passage normal human ovarian surface epithelial (HOSE) cells, human ovarian adenocarcinoma cell lines (CaOV-3, NIH:OVCAR-3, SK-OV-3 and SW626) and surgical specimens of human serous cystadenocarcinomas. We hypothesized that GJIC and connexin expression would be decreased in neoplastic cells and would be increased by cAMP and retinoic acid. Cultured HOSE cells exhibited extensive fluorescent dye-coupling and connexin43 (Cx43) expression; other connexins were not detected. The ovarian adenocarcinoma cell lines had little dye-coupling or connexin expression. Deletions and rearrangements of the Cx43 gene were not detected by Southern blotting in the carcinoma lines. N6, 2'-O dibutyryladenosine 3',5'-cyclic monophosphate and all-trans-retinoic acid inhibited cell proliferation, but did not enhance GJIC or Cx43 expression. Surface epithelial cells of benign ovaries expressed Cx43, but this expression was barely detectable in ovarian serous cystadenocarcinomas. Thus, normal HOSE cells had extensive GJIC and Cx43 expression whereas ovarian carcinoma cells had less and cAMP and retinoic acid did not change these, although both agents inhibited cell growth. PMID- 10383915 TI - Chemoprevention of tobacco smoke-induced lung tumors in A/J strain mice with dietary myo-inositol and dexamethasone. AB - Male A/J strain mice were fed AIN-76A diet supplemented with myo inositol/dexamethasone (10 g and 0.5 mg/kg diet) or acetylsalicylic acid (300 mg/kg) and exposed for 5 months to a mixture of sidestream and mainstream cigarette smoke at a concentration of 132 mg total suspended particulates/m3. After tobacco smoke exposure, they were allowed to recover for another 4 months in filtered air. In the animals fed AIN-75A diet alone or acetylsalicylic acid, the average number of tumors/lung was 2.1, whereas in the animals given the myo inositol/dexamethasone diet, the average lung tumor multiplicity was 1.0 (P < 0.05). In animals exposed to filtered air, lung tumor multiplicities were 0.6 for animals fed AIN-76A or myo-inositol/dexamethasone and 1.2 for animals fed acetylsalicylic acid. It was concluded that the combination of myo-inositol and dexamethasone constitutes an effective chemopreventive regimen against tobacco smoke-induced lung tumorigenesis. PMID- 10383916 TI - The effect of connexin32 null mutation on hepatocarcinogenesis in different mouse strains. AB - Connexin32 (Cx32) is the major gap junctional protein in mouse liver. We have shown recently that the formation of liver tumours in Cx32-deficient mice is strongly increased in comparison with control wild-type mice, demonstrating that the deficiency in gap junctional communication has an enhancing effect on hepatocarcinogenesis. We have now compared the effect of Cx32 deficiency on liver carcinogenesis in two strains of mice with differing susceptibility to hepatocarcinogenesis. Heterozygous Cx32(+/-) females were crossed with male Cx32 wild-type C57BL/6J (low susceptibility) or C3H/He (high susceptibility) mice. Since the Cx32 gene is located on the X-chromosome, the resulting F1 males segregated to the genotypes Cx32(Y/+) and Cx32(Y/-). Genotyping was performed by PCR-analysis using tail-tip DNA. Weanling male mice were i.p. injected with a single dose of N-nitrosodiethylamine and were killed 16, 21 or 26 weeks later. The number, volume fraction and size distribution of precancerous liver lesions characterized by a deficiency in the marker enzyme glucose-6-phosphatase were quantitated. The results demonstrate that Cx32 deficiency only slightly affects the number of enzyme-altered lesions, but strongly enhances their growth, both in the resistant and the susceptible mouse strain, suggesting that decreased intercellular communication results in tumour promoting activity irrespective of the genetic background of the mouse strain used. Since Cx32-deficient C3H/He hybrids were approximately 5-10 times more sensitive than C3H/He hybrids with an intact Cx32 gene, this mouse strain may prove very useful for toxicological screening purposes. PMID- 10383917 TI - Sertraline N-demethylation is catalyzed by multiple isoforms of human cytochrome P-450 in vitro. AB - Sertraline, a new antidepressant of the selective serotonin reuptake inhibitor class, is extensively metabolized to desmethylsertraline in humans. We identified the cytochrome P-450 (CYP) isoforms involved in sertraline N-demethylation using pooled human liver microsomes and cDNA-expressed CYP isoforms. Eadie-Hofstee plots for the sertraline N-demethylation in human liver microsomes were monophasic. The estimated Michaelis-Menten kinetic parameters were: KM = 18.1 +/- 2.0 microM, Vmax = 0.45 +/- 0.03 nmol/min/mg of protein, and Vmax/KM = 25.2 +/- 4.3 microl/min/mg of protein. At the substrate concentration of 20 microM, which approximated the apparent KM value, sulfaphenazole (CYP2C9 inhibitor) and triazolam (CYP3A substrate) reduced the N-demethylation activities by 20 to 35% in human liver microsomes, whereas the inhibition induced by mephenytoin (CYP2C19 substrate) or quinidine (CYP2D6 inhibitor) was marginal. The anti-CYP2B6 antibody inhibited the sertraline N-demethylation activities by 35%. Sertraline N demethylation activities were detected in all cDNA-expressed CYP isoforms studied. In particular, CYP2C19, CYP2B6, CYP2C9-Arg, CYP2D6-Val, and CYP3A4 all showed relatively high activity. When the contributions of CYP2D6, CYP2C9, CYP2B6, CYP2C19, and CYP3A4 were estimated from the Vmax/KM of cDNA-expressed CYP isoforms and from their contents in pooled human liver microsomes, the values were found to be 35, 29, 14, 13, and 9%, respectively. The results suggest that at least five isoforms of CYP (CYP2B6, CYP2C9, CYP2C19, CYP2D6, CYP3A4) are involved in the sertraline N-demethylation in human liver microsomes and that the contribution of any individual isoform does not exceed 40% of overall metabolism. Therefore, concurrent administration of a drug that inhibits a specific CYP isoform is unlikely to cause a marked increase in the plasma concentration of sertraline. PMID- 10383918 TI - Reductive metabolism In vivo of trans-4-phenyl-3-buten-2-one in rats and dogs. AB - The reductive metabolism in vivo of a flavoring additive, trans-4-phenyl-3-buten 2-one (PBO; trans-methyl styryl ketone) was investigated in rats and dogs. In both species, the double bond-reduced product, 4-phenyl-2-butanone (PBA), was detected by HPLC as the predominant species in blood after i.v. administration of PBO. PBA detected in rat blood was identified by comparison to the authentic sample. In contrast, the carbonyl-reduced product, trans-4-phenyl-3-buten-2-ol (PBOL) was also detected as a minor metabolite of PBO in both species. The area under the curve of PBOL in rat blood was only 3% of that of PBA. PBO was mutagenic in the Ames test using Salmonella typhimurium TA 100 when S-9 mix was added, but PBA and PBOL were not. It appears that PBO is mainly metabolized to PBA in vivo in rats and dogs as a detoxification pathway. PMID- 10383919 TI - Potent inhibition of cytochrome P-450 2D6-mediated dextromethorphan O demethylation by terbinafine. AB - Cytochrome P-450 (CYP) 2D6 is responsible for the biotransformation of over 35 pharmacologic agents. In the process of studying CYP2D6 we identified phenotype genotype discordance in two individuals receiving terbinafine. This prompted evaluation of the potential for terbinafine to inhibit CYP2D6 in vitro. Human hepatic microsomes and heterologously expressed CYP2D6 were incubated with terbinafine or quinidine and the formation of dextrorphan from dextromethorphan was determined by HPLC. Additionally, preliminary conformational analyses were conducted to determine the fit of terbinafine into a previously described pharmacophore model for CYP2D6 inhibitors. The apparent Km and Vmax of dextrorphan formation from four human hepatic microsome samples ranged from 5.8 to 6.8 microM and from 172 to 300 pmol/min/mg protein, respectively. Values of Km and Vmax in the heterologously expressed CYP2D6 system averaged 6.5 +/- 2.1 microM and 1342 +/- 147 pmol/min/mg protein, respectively. Terbinafine inhibited dextromethorphan O-demethylation with an apparent Ki ranging from 28 to 44 nM in human hepatic microsomes and averaging 22.4 +/- 0.6 nM for the heterologously expressed enzymes. Results of quinidine in these systems produced values for Ki ranging from 18 to 43 nM. Such strong inhibition of CYP2D6 by terbinafine would not have been predicted by the previously proposed pharmacophore model of CYP2D6 inhibitors based on molecular structure. Terbinafine is a potent inhibitor of CYP2D6 with apparent Ki values well below plasma and tissue concentrations typically achieved during a therapeutic course. This agent needs to be evaluated in vivo to determine the impact of CYP2D6 inhibition by terbinafine on the metabolism of concomitantly administered CYP2D6 substrates. PMID- 10383921 TI - Covalent sequestration of phosphoramide mustard by metallothionein--an in vitro study. AB - Acquired drug resistance is one of the most important problems in cancer chemotherapy. One of the proposed mechanisms for these phenomena is the sequestration of alkylating agents by metallothionein in vivo. This research shows that metallothionein can covalently sequester phosphoramide mustard, the active form of cyclophosphamide in vitro. On-line electrospray mass spectrometry reveals that it is phosphoramide, not nornitrogen mustard that alkylates metallothionein, although the metallothionein/nornitrogen mustard adduct was isolated as the major adduct. Tandem mass spectrometric experiments were performed on an isolated drug-modified tryptic peptide. The alkylation occurred predominantly at Cys48 of metallothionein. These results provide further evidence that overexpression of metallothionein can detoxify the active form of the drugs. PMID- 10383920 TI - An in vitro study on the metabolism and possible drug interactions of rokitamycin, a macrolide antibiotic, using human liver microsomes. AB - This in vitro study was designed to identify the enzyme(s) involved in the two major metabolic pathways of rokitamycin [formations of leucomycin A7 (LMA7) from rokitamycin and of leucomycin V (LMV) from LMA7] and to assess possible drug interactions using human liver microsomes. Formation of LMA7 or LMV was NADPH independent. Anti-rat NADPH cytochrome P-450 (CYP) reductase serum, specific inhibitors, or substrates of CYP isoforms showed no effects on the formation of LMA7 or LMV. The mean Vmax and Vmax/Km for the formation of LMA7 from rokitamycin were much greater (P <.01) than those for the formation of LMV from LMA7. Two esterase inhibitors, bis-nitro-phenylphosphate and physostigmine (100 microM), inhibited the formation of LMA7 or LMV by more than 85%, whereas no appreciable inhibition occurred by several substrates of carboxylesterase (EC 3.1.1.1). Except the moderate inhibition produced by promethazine and terfenadine, theophylline, mequitazine, chlorpheniramine, and diphenhydramine showed little or no inhibition for the formation of LMA7 or LMV. Rokitamycin, LMA7, LMV, erythromycin, and clarithromycin (up to 500 microM) had no appreciable inhibition for CYP1A2-, 2C9-, and 2D6-mediated catalytic reactions. However, rokitamycin, LMA7, erythromycin, and clarithromycin inhibited the CYP3A4-catalyzed triazolam alpha-hydroxylation with IC50 (Ki) values of 5.8 (2.0), 40, 33 (20), and 56 (43) microM, respectively. It is concluded that the formations of LMA7 from rokitamycin and of LMV from LMA7 are catalyzed mainly by human esterase enzyme [possibly cholinesterase (EC3.1.1.8)]. However, whether rokitamycin would inhibit the CYP3A-mediated drug metabolism in vivo requires further investigations in patients. PMID- 10383922 TI - Inhibitory effects of azelastine and its metabolites on drug oxidation catalyzed by human cytochrome P-450 enzymes. AB - Azelastine, an antiallergy and antiasthmatic drug, has been reported to be metabolized mainly to desmethylazelastine and 6-hydroxyazelastine in mammals. In the present study, the inhibitory effects of azelastine and its two metabolites on human cytochrome P-450 (CYP) isoform-dependent reactions were investigated to predict the drug interactions of azelastine using microsomes from human B lymphoblast cells expressing CYP. The specific activities for human CYP isoforms included: 7-ethoxyresorufin O-deethylation (CYP1A1), phenacetin O-deethylation (CYP1A2), coumarin 7-hydroxylation (CYP2A6), 7-benzyloxyresorufin O-dealkylation (CYP2B6), S-warfarin 7-hydroxylation (CYP2C9), S-mephenytoin 4'-hydroxylation (CYP2C19), bufuralol 1'-hydroxylation (CYP2D6), chlorzoxazone 6-hydroxylation (CYP2E1), and testosterone 6beta-hydroxylation (CYP3A4). In almost all the activities, desmethylazelastine exhibited stronger inhibition than azelastine and 6-hydroxyazelastine. Desmethylazelastine, but not azelastine and 6 hydroxyazelastine, uncompetitively inhibited CYP2B6 activity (Ki = 32.6 +/- 4.8 microM). Azelastine, desmethylazelastine, and 6-hydroxyazelastine competitively inhibited CYP2C9 activity (Ki = 13. 9 +/- 1.8, 15.0 +/- 3.1, and 17.0 +/- 4.1 microM, respectively), CYP2C19 activity (Ki = 21.9 +/- 2.2, 7.3 +/- 1.6, and 9.3 +/- 1.6 microM, respectively), and CYP2D6 activity (Ki = 1.2 +/- 0.1, 1.5 +/- 0.2, and 3.0 +/- 0.5 microM, respectively). Azelastine and desmethylazelastine competitively inhibited CYP3A4 activity (Ki = 23. 7 +/- 4.6 and 13.2 +/- 2.3 microM). 6-Hydroxyazelastine interfered with the determination of testosterone 6beta-hydroxylation by HPLC. CYP1A2, CYP2A6, and CYP2E1 activities were not significantly inhibited by azelastine and the two metabolites. Among the human CYPs tested, the inhibitory effects of azelastine and its two metabolites were the most potent on human CYP2D6. In consideration of the Ki values and the concentration of azelastine and desmethylazelastine in human livers after chronic oral administration of azelastine, the possibility of in vivo drug interaction of azelastine and other drugs that are mainly metabolized by CYP2D6 was suggested although it might not cause critical side effects. The inhibition of CYP2C9, CYP2C19, and CYP3A4 activity by azelastine and its two metabolites might be clinically insignificant. PMID- 10383923 TI - Specific dehydrogenation of 3-methylindole and epoxidation of naphthalene by recombinant human CYP2F1 expressed in lymphoblastoid cells. AB - 3-Methylindole (3MI) is a naturally occurring pulmonary toxin that requires metabolic activation. Previous studies have shown that 3MI-induced pneumotoxicity resulted from cytochrome P-450-catalyzed dehydrogenation of 3MI to an electrophilic methylene imine (3-methyleneindolenine), which covalently bound to cellular macromolecules. Multiple cytochrome P-450s are capable of metabolizing 3MI to several different metabolites, including oxygenated products. In the present study, the role of human CYP2F1 in the metabolism of 3MI was examined to determine whether it catalyzes dehydrogenation rather than hydroxylation or ring oxidation. Metabolism was examined using microsomal fractions from human lymphoblastoid cells that expressed the recombinant human CYP2F1 P-450 enzyme. Expression of CYP2F1 in the lymphoblastoid cells proved to be an appropriate expression system for this enzyme. Products were analyzed using HPLC and the mercapturate, 3-[(N-acetylcystein-S-yl)methyl]indole, of the reactive intermediate was identified and quantified. Product analysis showed that human CYP2F1 efficiently catalyzed the dehydrogenation of 3MI to the methylene imine without detectable formation of indole-3-carbinol or 3-methyloxindole. High substrate concentrations of 3MI strongly inhibited production of the dehydrogenated product, a result that may indicate the existence of mechanism based inhibition of CYP2F1 by 3MI. Recombinant CYP2F1 demonstrated remarkable selectivity for the bioactivation of 3MI to the putative dehydrogenated reactive electrophile. Bioactivation of naphthalene to its pneumotoxic epoxide by CYP2F1 was also demonstrated. PMID- 10383924 TI - Characterization of human small intestinal cytochromes P-450. AB - Human small intestine epithelial cells (enterocytes) provide the first site for cytochrome P-450 (CYP)-catalyzed metabolism of orally ingested xenobiotics. The CYP composition of enterocytes could thus affect the potential toxicity or therapeutic efficacy of xenobiotics by modifying systemic uptake. We have characterized human enterocyte CYP composition to enable assessment of its functional roles. An isolation method for enterocytes from human small intestine was developed using EDTA buffer-mediated elution. Villous enterocytes were isolated in high yield, separated from crypt cells. Reverse transcriptase polymerase chain reaction of total RNA from enterocytes revealed that CYP1A1, 1B1, 2C, 2D6, 2E1, 3A4, and 3A5 mRNA were expressed, but only CYP2C and 3A4 were detectable by Western immunoblotting in enterocyte microsomes from 10 human small intestines, whereas CYP1A1 was weakly detectable in two of eight intestines tested. Microsomal protein content decreased markedly along the small intestine from the duodenum to the ileum, whereas total CYP content and CYP3A4 erythromycin N-demethylase activity increased slightly in progressing from the duodenum to the jejunum and then decreased markedly toward the ileum. Levels of CYP3A4 and 2C protein did not decrease in concert as a function of length along the intestine distally. Maximal CYP content for the 10 intestines varied from 0.06 to 0.18 nmol/mg microsomal protein and maximal CYP3A4 erythromycin N-demethylase activity varied from 0.30 to 0.76 nmol/min/mg microsomal protein. In conclusion, CYP3A4 is the major form of CYP expressed in human small intestine enterocytes, CYP3A5 expression was not detected, CYP2C and, in some intestines, CYP1A1 were expressed. The highest metabolic activity occurred in the proximal intestine. PMID- 10383925 TI - Absorption, excretion, and metabolism of the endothelin receptor antagonist bosentan in healthy male subjects. AB - The absorption, excretion, and metabolism of the endothelin receptor antagonist bosentan was investigated in healthy male subjects by administration of 14C labeled compound. Four subjects received a single oral dose of 500 mg of bosentan (3.7 MBq), and four other subjects received a single i.v. dose of 250 mg of bosentan (3.7 MBq). Radioactivity and concentrations of bosentan and its metabolites were measured in plasma, urine, and feces samples. More than 97% of drug-related material was recovered on average within 3.5 days after oral dosing and within 5 days after i.v. dosing. More than 90% of radioactivity was found in feces after both oral and i.v. dosing. Most of the radioactivity in urine and feces represented bosentan and three metabolites. Ro 48-5033, the major metabolite in plasma, urine, and feces, is the result of hydroxylation at the t butyl group of bosentan. The two other metabolites Ro 47-8634 and Ro 64-1056 represent minor metabolite species. Ro 47-8634 is the product of O-demethylation of the phenolic methyl ester, and Ro 64-1056 is generated by both demethylation and hydroxylation. The radioactivity in plasma could almost entirely be attributed to bosentan and the two metabolites Ro 48-5033 and Ro 47-8634, whereby both metabolites exhibited much lower plasma levels than bosentan. Hepatic metabolism followed by biliary excretion of the metabolites apparently represents the major pathway of elimination for bosentan in humans. PMID- 10383926 TI - Prediction of the systemic exposure to oral 9-amino-20(S)-camptothecin using single-sample analysis. AB - The purpose of this study was to develop and validate limited-sampling strategies for prediction of the area under the plasma-concentration time curves (AUCs) of the lactone and total (i. e., lactone plus carboxylate) forms of the novel topoisomerase-I inhibitor 9-amino-20(S)-camptothecin (9-AC). Complete pharmacokinetic curves for both drug species were obtained from 32 patients who received the drug orally in a clinical phase I setting at dose levels ranging from 0.25 to 1.10 mg/m2. The concentrations of the lactone and carboxylate forms of 9-AC in plasma were measured by HPLC. Using data from 20 randomly selected patients, forward-stepwise multivariate regression analysis was used to generate modeling strategies incorporating data from one, two, or three plasma samples. The simultaneous optimal prediction of both 9-AC lactone and 9-AC total AUCs was obtained with sample time points at 0.33, 3.0, and 11.0 h after drug dosing. Validation of the models on an independent data set comprising data of the remaining 12 patients demonstrated that 9-AC lactone and 9-AC total AUCs could be predicted sufficiently unbiased and precise using one and two time points: [AUC (ng. h/ml) = 7.103*C3 + 4.333] for 9-AC lactone and [AUC (ng. h/ml) = 9.612*C3 + 13.77*C11 - 44.11] for 9-AC total, where C3 and C11 represent the 9-AC plasma concentrations in ng/ml at 3 and 11 h after drug dosing. Application of the proposed models will be valuable in the determination of 9-AC population pharmacokinetics and permits treatment optimization for patients on the basis of individual pharmacokinetic characteristics through restricted drug monitoring in clinical routines. PMID- 10383927 TI - Intracoronary and intravenous administration of basic fibroblast growth factor: myocardial and tissue distribution. AB - Therapeutic angiogenesis using various heparin-binding growth factors is a promising treatment for ischemic heart disease. Single dose intracoronary (IC) or i.v. delivery are most practical for clinical use. This study was designed to investigate the myocardial and tissue deposition of basic fibroblast growth factor (bFGF) after IC and i.v. administration in normal and chronically ischemic animals. Twenty-four Yorkshire pigs were used (12 normal and 12 ischemic animals) with IC and i.v. administration of 125I-bFGF (25 microCi) combined with cold bFGF (30 microg) and heparin (3 mg). Tissue and myocardial distribution was determined at 1 and 24 h by measuring 125I-bFGF specific activity and by organ and light level autoradiography. The liver accounted for the majority of 125I-bFGF activity at 1 h (37.6 +/- 17.1% for IC and 42.1 +/- 17.7% for i.v. delivery), with a reduction to 2.8 +/- 1.5% for IC and 1.5 +/- 0.9% for i.v. delivery by 24 h. Total cardiac specific activity at 1 h was 0.88 +/- 0.89% for IC and 0.26 +/- 0.08% for i.v. administration (p =.12) and decreased to 0.05 +/- 0.04% (p =.05, versus 1 h) and 0. 04 +/- 0.01% (p <.001, versus 1 h) at 24 h, respectively. IC but not i.v. delivery resulted in higher deposition in ischemic than normal myocardium. IC delivery resulted in enhanced bFGF deposition only in myocardial territories subtended by the infused artery. Intravenous delivery compares favorably with IC delivery with a 3- to 4-fold reduction in myocardial deposition at 1 h and with similar solid organ deposition. The less invasive nature of i.v. delivery, its potential for repeat administration, and its applicability to a larger population may offset its resultant reduced myocardial deposition. Efficacy studies are ongoing. PMID- 10383928 TI - Effects of a potent and specific P-glycoprotein inhibitor on the blood-brain barrier distribution and antinociceptive effect of morphine in the rat. AB - Previous data suggest that the analgesic effect of morphine may be modulated by P glycoprotein (P-gp) inhibition. The effects of the P-gp inhibitor GF120918 on brain distribution and antinociceptive effects of morphine were examined in a rat cerebral microdialysis model. Pretreatment with GF120918 increased both the area under the concentration-time curve of unbound morphine in brain extracellular fluid (ECF) and morphine-associated antinociception. The area under the concentration-time curve ratio for unbound morphine in brain ECF versus unbound morphine in blood was significantly higher in GF120918-treated rats compared with control rats (1.21 +/- 0.34 versus 0.47 +/- 0.05, respectively; p <.05). Modulation of morphine brain-blood distribution was confirmed by quantitating brain tissue morphine in a separate group of rats; GF120918 increased the brain tissue:serum concentration ratio approximately 3-fold. The half-life of unbound morphine in brain ECF was approximately 3-fold longer in GF120918-treated rats compared with controls (p <.05). The fraction unbound of morphine in whole blood was not altered significantly in the presence of GF120918 (0.651 +/- 0.039) as compared with controls (0.662 +/- 0.035). Concentrations of unbound morphine-3 glucuronide in blood and brain ECF were increased in GF120918-treated rats versus controls. An integrated pharmacokinetic/pharmacodynamic model was developed to characterize the unbound blood and brain ECF morphine concentration profiles and concentration-effect relationships. The results of this study indicate that alteration of morphine antinociception by a potent P-gp inhibitor appears to be mediated at the level of the blood-brain barrier. PMID- 10383929 TI - Metabolism of diallyl disulfide by human liver microsomal cytochromes P-450 and flavin-containing monooxygenases. AB - The metabolism of diallyl disulfide (DADS), a garlic sulfur compound, was investigated in human liver microsomes. Diallyl thiosulfinate (allicin) was the only metabolite observed and its formation followed Michaelis-Menten kinetics with a Km = 0.61 +/- 0.2 mM and a Vmax = 18.5 +/- 4.2 nmol/min/mg protein, respectively (mean +/- S.E. M., n = 4). Both flavin-containing monooxygenase and the cytochrome P-450 monooxygenases (CYP) were involved in DADS oxidation, but the contribution of CYP was predominant. The cytochrome P-450 isoforms involved in this metabolism were investigated using selective chemical inhibitors, microsomes from cells expressing recombinant CYP isoenzymes, and studying the correlation of the rate of DADS oxidation with specific monooxygenase activities of human liver microsomes. Diethyldithiocarbamate and tranylcypromine inhibited allicin formation, whereas other specific inhibitors had low or no effect. Most of the different human microsomes from cells expressing CYP were able to catalyze this reaction, but CYP2E1 showed the highest affinity with a substantial activity. Furthermore, allicin formation by human liver microsomes was correlated with p-nitrophenol hydroxylase activity, a marker of CYP2E1, and tolbutamide hydroxylase activity, a marker of CYP2C9. Among these approaches only CYP2E1 was identified in each case, which suggested that DADS is preferentially metabolized to allicin by CYP2E1 in human liver. However the minor participation of other CYP forms and flavin-containing monooxygenases is likely. PMID- 10383930 TI - Comparison of human and rat metabolism of molinate in liver microsomes and slices. AB - Molinate undergoes oxidative metabolism forming either ring-hydroxylated metabolites or molinate sulfoxide. Our previous studies strongly implicated the sulfoxidation pathway in molinate-induced testicular toxicity. The present study compares the metabolic capability of rat and human liver microsomes and slices to form either nontoxic ring-hydroxylated metabolites of molinate or the toxic metabolites derived from the sulfoxidation of molinate. Km and Vmax values indicate that sulfoxidation would be the preferred high-dose pathway whereas hydroxylation would predominate at low dose levels in both species. Examination of phase II metabolism of molinate in liver slices reveals greater detoxification of molinate sulfoxide by glutathione conjugation in humans with rats forming less conjugate. Oxidative metabolism of molinate in both rats and humans appears to be mediated by cytochrome P-450 and not flavin monooxygenases as indicated by the use of metabolic inhibitors. Overall, the metabolism of molinate would be via the nontoxic hydroxylation pathway in both species at low doses whereas at high doses, where sulfoxidation would predominate, the human is more capable than the rat to detoxify via glutathione conjugation. PMID- 10383931 TI - Identification of urinary metabolites of isoprene in rats and comparison with mouse urinary metabolites. AB - Isoprene, a major commodity chemical used in production of polyisoprene elastomers, has been shown to be carcinogenic in rodents. Similar to findings for the structurally related compound butadiene, mice are more susceptible than rats to isoprene-induced toxicity and carcinogenicity. Although differences in uptake, and disposition of isoprene in rats and mice have been described, its in vivo biotransformation products have not been characterized in either species. The purpose of these studies was to identify the urinary metabolites of isoprene in Fischer 344 rats and compare these metabolites with those formed in male B6C3F1 mice. After i.p. administration of 64 mg [14C]isoprene/kg to rats and mice, isoprene was excreted unchanged in breath ( approximately 50%) or as urinary metabolites ( approximately 32%). In rats isoprene was primarily excreted in urine as 2-hydroxy-2-methyl-3-butenoic acid (53%), 2-methyl-3-buten-1,2-diol (23%), and the C-1 glucuronide conjugate of 2-methyl-3-buten-1,2-diol (13%). These metabolites are consistent with preferential oxidation of isoprene's methyl substituted vinyl group. No oxidation of the unsubstituted vinyl group was observed. In addition to the isoprene metabolites found in rat urine, mouse urine contained numerous other isoprene metabolites with a larger percentage (25%) of total urinary radioactivity associated with an unidentified, polar fraction than in the rat (7%). Unlike butadiene, there was no evidence that glutathione conjugation played a significant role in the metabolism of isoprene in rats. Because of the unidentified metabolites in mouse urine, involvement of glutathione in the metabolism of isoprene in mice cannot be delineated. PMID- 10383932 TI - Molecular mechanisms in the TCR (TCR alpha beta-CD3 delta epsilon, gamma epsilon) interaction with zeta 2 homodimers: clues from a 'phenotypic revertant' clone. AB - The association between the TCRalphabeta-CD3gammaepsilondeltaepsilon hexamers and zeta2 homodimers in the endoplasmic reticulum (ER) constitutes a key step in TCR assembly and export to the T cell surface. Incompletely assembled TCR-CD3 complexes are degraded in the ER or the lysosomes. A previously described Jurkat variant (J79) has a mutation at position 195 on the TCR Calpha domain causing a phenylalanine to valine exchange. This results in a lack of association between TCRalphabeta-CD3gammaepsilondeltaepsilon hexamers and zeta2 homodimers. Two main hypotheses could explain this phenomenon in J79 cells: TCR-CD3 hexamers may be incapable of interacting with zeta2 due to a structural change in the TCR Calpha region; alternatively, TCR-CD3 hexamers may be incapable of interacting with zeta2 due to factors unrelated to either molecular complex. In order to assess these two possibilities, the TCR-CD3 membrane-negative J79 cells were treated with ethylmethylsulfonate and clones positive for TCR membrane expression were isolated. The characterization of the J79r58 phenotypic revertant cell line is the subject of this study. The main question was to assess the reason for the TCR re-expression. The TCR on J79r58 cells appears qualitatively and functionally equivalent to wild-type TCR complexes. Nucleotide sequence analysis confirmed the presence of the original mutation in the TCR Calpha region but failed to detect compensatory mutations in alpha, beta, gamma, delta, epsilon or zeta chains. Thus, mutated J79-TCR-CD3 complexes can interact with zeta2 homodimers. Possible mechanisms for the unsuccessful TCR-CD3 interaction with zeta2 homodimers are presented and discussed. PMID- 10383934 TI - Evidence that human CD8+CD45RA+CD27- cells are induced by antigen and evolve through extensive rounds of division. AB - We recently showed that circulating human CD8(+) effector cells have a CD45RA+CD27(-) membrane phenotype. In itself this phenotype appeared to pose a paradox: CD45RA, a marker expressed by unprimed cells, combined with absence of CD27, characteristic for chronically stimulated T cells. To investigate whether differentiation towards the CD45RA+CD27(-) phenotype is dependent on antigenic stimulation and involves cellular division, TCR Vbeta usage and telomeric restriction fragment (TRF) length were analyzed within distinct peripheral blood CD8(+) subsets. FACS analysis showed that the TCR Vbeta repertoire of CD8(+)CD45RA+CD27(-) cells differed significantly from that of unprimed CD8(+)CD45RA+CD27(+) cells. Moreover, in two out of six individuals large expansions of particular Vbeta families were observed in the CD8(+)CD45RA+CD27(-) subset. CDR3 spectrotyping and single-strand confirmation analysis revealed that within the CD8(+)CD45RA+CD27(-) population most of the 22 tested Vbeta families were dominated by oligoclonal expansions. The mean TRF length was found to be 2.3+/-1.0 kb shorter in the CD8(+)CD45RA+CD27(-) subset compared with the unprimed CD8(+)CD45RA+CD27(+) population, but did not differ substantially from that of memory type, CD8(+)CD45RA-CD27(+) T cells. These findings indicate that the CD8(+)CD45RA+CD27(-) cytotoxic effector population consists of antigen induced, clonally expanded cells and confirm that the expression of CD45RA is not a strict marker of antigen non-experienced T cells. PMID- 10383935 TI - Severe impairment of B cell function in lpr/lpr mice expressing transgenic Fas selectively on B cells. AB - Transgenic lpr/lpr mice expressing functional Fas selectively on B cells were produced in an attempt to elucidate the role of Fas on B cells in the regulation of autoantibody production. The homozygous lpr/lpr mice carrying the transgene did not produce anti-double-stranded DNA antibodies throughout their lives, whereas the development of abnormal lpr T cells (double negative, B220(+)) was not suppressed. Further analyses, however, revealed that the expression of the transgenic Fas on B cells of lpr/lpr homozygous mice resulted in severe impairment of the B cell function. The defect was characterized by a decrease in the number of mature peripheral B cells, a reduction in the serum Ig level and the total failure of B cells to mount antibody responses to stimulations of T dependent as well as T-independent antigens. Such a defect was prominent only when the transgene was expressed on the lpr/lpr homozygous background. On the contrary, B cells of the transgenic lpr/lpr mice were shown to be capable of producing Ig when stimulated with anti-CD40 in the presence of IL-4 and IL-5. Furthermore, lpr/lpr T cells showed enhanced non-specific cytolytic activity. These observations suggested that the observed B cell defect was probably attributable to the destruction of activated B cells expressing transgenic Fas by aggressive lpr/lpr T cells. PMID- 10383933 TI - Expression pattern of notch1, 2 and 3 and Jagged1 and 2 in lymphoid and stromal thymus components: distinct ligand-receptor interactions in intrathymic T cell development. AB - The suggested role of Notch1 or its mutants in thymocyte differentiation and T cell tumorigenesis raises the question of how the different members of the Notch family influence distinct steps in T cell development and the role played by Notch ligands in the thymus. We report here that different Notch receptor-ligand partnerships may occur inside the thymus, as we observed differential expression of Notch1, 2 and 3 receptors, their ligands Jagged1 and 2, and downstream intracellular effectors hairy and Enhancer of Split homolog 1 (HES-1) and hairy and Enhancer of Split homolog 5 (HES-5), depending on ontogenetic stage and thymic cell populations. Indeed, while Jagged2 is expressed in both stromal cells and thymocytes, Jagged1 expression is restricted to stromal cells. Moreover, a differential distribution of Notch3, with respect to Notch1, was observed in distinct age-related thymocyte subsets. Finally, Notch3 was preferentially up regulated in thymocytes, following the induction of their differentiation by interaction with thymic epithelial cells expressing the cognate Jagged1 and 2 ligands, suggesting that, besides Notch1, Notch3 may also be involved in distinct steps of thymocyte development. Our results suggest that the Notch signaling pathway is involved in a complex interplay of T cell developmental stages, as a consequence of the heterogeneity and specific expression of members of the Notch receptor family and their cognate ligands, in distinct thymic cell compartments. PMID- 10383936 TI - Intranasal administration of peptide vaccine protects human/mouse radiation chimera from influenza infection. AB - Influenza virus is characterized by frequent and unpredictable changes of the surface glycoproteins which enable the virus to escape the immune system. Approved vaccines which consist of the whole virus or the surface glycoproteins fail to induce broad specificity protection. We have previously reported that a peptide-based experimental recombinant vaccine which includes conserved epitopes of B and T lymphocytes was efficient in mice, leading to cross-strain, long-term protection. In the present study, this approach was adapted for the design of a human vaccine, based on epitopes recognized by the prevalent HLAs. These epitopes were expressed in Salmonella flagellin and tested for their efficacy in human/mouse radiation chimera in which human peripheral blood mononuclear cells (PBMC) are functionally engrafted. The vaccinated mice demonstrated clearance of the virus after challenge and resistance to lethal infection. The production of virus-specific human antibodies was also higher in this group. Control groups of either non-vaccinated, or vaccinated mice which had not been engrafted with the human PBMC, did not exhibit the protective immune response. FACS analysis showed that most human cells in the transplanted mice are CD8(+) and CD4(+). Hence, it may be concluded: (i) that the protection involves cellular mechanisms, but is most probably accomplished without direct lysis of influenza-infected pulmonary cells by cytotoxic T lymphocytes, but rather via a cytokine-mediated mechanism, (ii) that the human/mouse radiation chimera model may be of some value in the investigation of new vaccines, as an additional tool prior to clinical trials, and (iii) that the synthetic recombinant vaccine can induce a response in the human immune system and confers protection against influenza infection. Further investigation is needed to establish the efficacy of such a peptide vaccine in human subjects. PMID- 10383937 TI - Soluble IL-6 receptor induces calcium flux and selectively modulates chemokine expression in human dermal fibroblasts. AB - Truncated forms of cytokine receptors have been regarded as modulators of the activity of their cognate ligands. In addition to inhibiting effects of their respective ligands, soluble receptors can also facilitate ligand-mediated signaling. Several studies have demonstrated that exogenous IL-6 in association with the soluble IL-6 receptor alpha (sIL-6Ralpha) can activate cells expressing the gp130 signal transducer lacking the specific, membrane-bound IL-6Ralpha. Since cell cultures of human dermal fibroblasts express high amounts of IL-6, we examined whether the addition of sIL-6Ralpha in association with endogenous IL-6 would be sufficient to stimulate these cells via gp130. As an early rapid signal we analyzed changes in intracellular free calcium concentrations ([Ca2+]i). Addition of sIL-6Ralpha induced an acute and transient increase in cytosolic free calcium concentrations in a dose-dependent fashion. This Ca2+-signal was abolished when cells were pretreated with anti-IL-6 or anti-gp130 antibodies. Using flow cytometric analysis we could demonstrate membrane-associated IL-6 and gp130, but not IL-6Ralpha on fibroblasts. We also analyzed MCP-1 and IL-8 expression as a response involved in the more recently recognized chemoattractant functions of fibroblasts, and found MCP-1 to be up-regulated, but not IL-8. These data suggest that sIL-6Ralpha binds to cell-associated, endogenous IL-6 produced by fibroblasts and this complex then activates the cells via gp130. This pathway of fibroblast activation by sIL-6Ralpha adds another dimension to the role of fibroblasts in the cytokine network. PMID- 10383938 TI - BALB/c.CBA/N mice carrying the defective Btk(xid) gene are resistant to pristane induced plasmacytomagenesis. AB - The X-chromosome from the CBA/N mouse which carries the defective Bruton's tyrosine kinase (Btk) allele (Xxid) has been introgressively backcrossed onto the plasmacytoma (PCT) induction-susceptible BALB/cAN. Inbred BALB/c.CBA/N-xid/xid (C.CBA/N) mice raised and maintained in our conventional colony were given three 0.5 ml injections of pristane and were highly refractory to PCT induction. Only one PCT was found among 59 mice followed for > or =300 days. Twenty mice were examined at day 200 for foci of plasma cells in the oil granuloma. Ten mice had small foci of plasma cells, most of which were plasmacytotic, embedded in the inflammatory oil granuloma. In one there were multiple foci, but most of the mice had only one or two foci. F1 hybrid XxidY males derived from CBA/N females crossed to BALB/cAnPt were also resistant to PCT induction, while heterozygous and homozygous XY males were susceptible. C.CBA/N mice can develop extensive mucosal plasma cells as well as plasma cell accumulations in oil granuloma tissue, but the precursors of these plasma cells do not give rise to PCT in genetically susceptible hosts. The failure of C.CBA/N mice to develop PCT is probably due to the elimination of B cell clones that can be perpetuated by repeated exposure to thymus-independent type 2 antigens. PMID- 10383939 TI - Collagen-induced arthritis development requires alpha beta T cells but not gamma delta T cells: studies with T cell-deficient (TCR mutant) mice. AB - Collagen type II (CII)-induced arthritis (CIA) in mice is a model for rheumatoid arthritis (RA) in which the role of T lymphocytes remains controversial. To clarify this, we have bred a targeted gene deletion of TCR beta or delta loci into two mouse strains susceptible to CIA, the B10.Q and DBA/1 strains. The TCRbeta-/- mice lacked alphabeta T cells, which was compensated by an expansion of B cells, gammadelta T cells and NK cells. The beta-/- mice, but not control beta+/- littermates, were completely resistant to CIA. The production of anti-CII IgG antibodies was also abolished in beta-/- mice, revealing a strict alphabeta T cell dependency. In contrast, beta-/- mice produced reduced, but significant, anti-CII IgM titers after immunization with either CII or ovalbumin, indicating a multispecificity for these alphabeta T cell-independent IgM antibodies. The TCRdelta-/- mice lacked gammadelta T cells but had no other significant changes in lymphocyte or monocyte subsets. The cytokine response (IL-2, IL-4, IL-10 and IFN-gamma) in delta-/- mice, quantified by flow cytometry staining of mitogen stimulated lymphocytes, was indistinguishable from normal mice. Likewise, no statistically significant differences were observed in CIA between mice lacking gammadelta T cells and control littermates, considering arthritis incidence, day of disease onset, maximum arthritic score, anti-CII IgG titers and disease course. We conclude that alphabeta T cells are necessary for CIA development and for an IgG response towards CII, whereas gammadelta T cells are neither necessary nor sufficient for development of CIA. PMID- 10383940 TI - STAT1 activation during monocyte to macrophage maturation: role of adhesion molecules. AB - Human monocytes isolated from peripheral blood of healthy donors show a time dependent differentiation into macrophages upon in vitro cultivation, closely mimicking their in vivo migration and maturation into extravascular tissues. The mediator(s) of this maturation process has not been yet defined. We investigated the involvement of signal transducers and activators of transcription (STAT) factors in this phenomenon and reported the specific, time-dependent, activation of STAT1 protein starting at day 0/1 of cultivation and maximally expressed at day 5. STAT1 activity was evident on the STAT binding sequences (SBE) present in the promoters of genes which are up-regulated during monocyte to macrophage maturation such as FcgammaRI and ICAM-1, and in the promoter of the transcription factor IFN regulatory factor-1. Moreover, the effect of cell adhesion to fibronectin or laminin was studied to investigate mechanisms involved in STAT1 activation. Compared with monocytes adherent on plastic surfaces, freshly isolated cells allowed to adhere either to fibronectin- or laminin-coated flasks exhibited an increased STAT1 binding activity both in control and in IFN-gamma treated cells. The molecular events leading to enhanced STAT1 activation and cytokine responsiveness concerned both Y701 and S727 STAT1 phosphorylation. Exogenous addition of transforming growth factor-beta, which exerts an inhibitory effect on some monocytic differentiation markers, inhibited macrophage maturation, integrin expression and STAT1 binding activity. Taken together these results indicate that STAT1 plays a pivotal role in the differentiation/maturation process of monocytes as an early transcription factor initially activated by adherence and then able to modulate the expression of functional genes, such as ICAM-1 and FcgammaRI. PMID- 10383941 TI - CD44 signaling through p56lck involves lateral association with CD4 in human CD4+ T cells. AB - CD44 is a family of mucin-like membrane proteins generated by alternative splicing of several exons, and participate in T cell adhesion and activation. CD44-mediated signaling involves activation of p56(lck) and leads to ZAP-70 phosphorylation. The aim of the present study was to identify the signaling pathways that follow CD44-triggered ZAP-70 phosphorylation and the molecular mechanisms underlying the CD44 interaction with p56(lck). We found that CD44 cross-linking by mAb in CD4(+) peripheral blood T cells promotes formation of a trimeric complex of Grb2, phospholipase (PLC)-gamma1 and a 36-38 kDa phosphoprotein, and the activation of PLC-gamma1. The amount of inositol triphosphate and the time kinetics of its generation were comparable to those following CD3 cross-linking. Co-capping, co-immunoprecipitation and fluorescence resonance energy transfer experiments showed that CD44 associates with CD4 and CD3 on the cell surface. This association suggests functional interplay between the CD4-TCR complex and CD44. In line with this possibility, we found that CD4 triggering by gp120, a natural ligand of CD4, potentiates CD44-mediated adhesion to hyaluronic acid. Moreover, Ca2+ mobilization induced by CD44 cross-linking by mAb was higher in a subclone of the HUT78 cell line expressing CD4 than in a non expressing subclone. PMID- 10383942 TI - Adjuvants that enhance priming of cytotoxic T cells to a Kb-restricted epitope processed from exogenous but not endogenous hepatitis B surface antigen. AB - Intramuscular (i.m.) or s.c. injection of plasmid DNA encoding hepatitis B small surface antigen (HBsAg) primes potent MHC I-restricted cytotoxic T lymphocyte (CTL) responses in H-2(d) (BALB/c) and H-2(b) (C57BL/6) mice. In contrast, i.m. or s.c. injection of exogenous HBsAg particles without adjuvants primes CTL responses in 'high responder' H-2(d) but not 'low responder' H-2(b) mice. We have shown that processing of exogenous but not endogenous HBsAg generates the Kb binding S208-215 peptide ILSPFLPL. This system allowed us to optimize conditions for stimulating murine CTL responses to exogenous antigen by identifying adjuvants that facilitate priming of Kb-restricted CTL by injecting recombinant HBsAg particles into 'low responder' H-2(b) mice. Synthetic oligodeoxynucleotides with immunostimulating sequences or the recombinant cytokine IL-12 efficiently enhanced priming of CTL to exogenous HBsAg. Hence, the adjuvanticity of DNA sequences that induce Th1 cytokines facilitate priming of MHC I-restricted T cell responses to exogenous antigen and are therefore of potential value in formulating vaccines designed to enhance CTL priming to exogenous antigen. PMID- 10383943 TI - Antigenic and immunogenic properties of totally synthetic peptide-based anti fertility vaccines. AB - In this study we describe the results of experiments in which a variety of totally synthetic luteinizing hormone releasing hormone (LHRH) vaccines were assembled and examined for their abilities to elicit antibody responses and induce sterility in mice. It is shown that totally synthetic vaccines consisting of a 15 residue-defined T cell epitope and the 10 residue LHRH epitope not only induced high titers of antibody but also induced sterility. This effect did not appear to correlate with antibody titer, antibody isotype or comparative antibody affinity, but may be related to the length of time for which antibodies are present to exert their influence. PMID- 10383944 TI - Enhancement of antigen-presenting cell surface molecules involved in cognate interactions by immunostimulatory DNA sequences. AB - Bacterial genomic DNA, plasmid DNA (pDNA) and synthetic oligodeoxynucleotides (ODN) containing immunostimulatory DNA sequences (ISS) have been proposed to foster a Th1 response via the release of type 1 cytokines from macrophages, dendritic cells, NK cells and B cells. In this study, we show that ISS-enriched DNA up-regulates a distinct profile of cell surface molecules on macrophages and B cells in vitro and in vivo. ISS-ODN and ISS-containing pDNA enhanced the expression of antigen presentation molecules (MHC class I and II), co-stimulatory molecules (B7-1, B7-2 and CD40), cytokine receptors (IFN-gamma receptor and IL-2 receptor), an adhesion molecule (ICAM-1) and an Fc receptor (Fcgamma receptor) on murine B cells or bone marrow-derived macrophages. The increased expression of these surface molecules is seen in purified cell populations and is largely independent of the effects of type 1 cytokines. Splenic antigen-presenting cells stimulated with ISS-ODN in vivo efficiently activate naive T cells and bias their differentiation toward a Th1 phenotype in vitro. Thus, the induction of both type 1 cytokines and a distinct profile of cell surface molecules contributes to the potent immunostimulatory effects of ISS-containing DNA on innate and adaptive immunity. PMID- 10383945 TI - Development of rat CD45+ 13-day-old fetal liver cells in SCID mouse fetal thymic organ cultures. AB - A phenotypic analysis of the lympho-hemopoietic cells which occur in the liver of 13-day-old fetal rats was achieved by flow cytometry in an attempt to further characterize the rat lymphoid progenitor cells. A small fraction of rat 13-day old fetal liver (r13FL) cells, which weakly expressed the leukocyte common antigen CD45, constituted a homogeneous Thy-1(hi), CD71(-), CD44(+), MHC class I+, CD43(+) cell subpopulation negative for CD45RC, CD3, TCRalphabeta, TCRgammadelta, CD2, CD5, CD4, CD8, CD25, CD28, NKR-P1a and sIg. On the contrary, the CD45(-) cells were a heterogeneous cell subset which expressed Thy-1, CD71 and CD44 at distinct levels. After MACS separation, the CD45(+) r13FL cells, but not the CD45(-) cell subset, in vitro repopulated 14-day-old SCID mouse fetal thymic lobes providing rat T cells, both TCRalphabeta and TCRgammadelta, NK cells, and thymic dendritic cells but not B lymphocytes. Interestingly, NKR P1a(lo) TCRalphabeta+ or TCRgammadelta+ cells developed in the xenogeneic cultures, and a rare CD4(+)CD8(+) double-positive subpopulation among the TCRgammadelta-expressing cells accumulated in the oldest cultures. These results are discussed from the double perspective of the nature of the precursor cells which colonize the fetal thymus and the relevance of the xenogeneic SCID mouse fetal thymic microenvironment for supporting rat lymphopoiesis. PMID- 10383947 TI - CD40 signaling induces B cell responsiveness to multiple members of the gamma chain-common cytokine family. AB - CD40 signaling induces B cell proliferative and differentiation responses that can be modulated by many different cytokines. Cytokines in the IL-2 receptor gamma chain (gammac)-common family are known to play an integral role in B cell development. Therefore, we investigated the possibility that CD40 signaling induced B cell responsiveness to multiple gammac-common cytokines and that individual gammac-common cytokines induced distinct B cell responses. B cells were isolated from lymphoid follicles of sheep Peyer's patches (PP) and co cultured with murine CD40 ligand (mCD40L). CD40 signaling induced PP B cell responsiveness to recombinant human IL-2, IL-4, IL-7 and IL-15. mCD40L-induced B cell growth was enhanced by combining IL-4 with a second gammac-common cytokine and sustained B cell growth required co-stimulation with IL-4 plus IL-2, IL-7 and IL-15. gammac-common cytokine responsiveness remained dependent upon CD40 signaling, and removal of mCD40L resulted in B cell differentiation and cell death. Similar proliferative responses to mCD40L and gammac-common cytokines were observed for both immature (ileal) and mature (jejunal) PP B cells. Finally, the capacity of CD40-activated B cells to respond to multiple gammac-common cytokines was analyzed with individual PP B cell clones. All B cell clones displayed similar proliferative responses to IL-2 but quantitatively different responses to IL-4, IL-7 and IL-15. The biological significance of B cell responsiveness to multiple gammac-common cytokines is discussed. PMID- 10383946 TI - Suppressive versus stimulatory effects of allergen/cholera toxoid (CTB) conjugates depending on the nature of the allergen in a murine model of type I allergy. AB - Recent reports have demonstrated that feeding small amounts of antigen conjugated to the B subunit of cholera toxin (CTB) suppress immune responses in experimental models of certain Th1-based autoimmune diseases. We have established a model of aerosol sensitization leading to Th2-mediated allergic immune responses in BALB/c mice. In the present study two different antigens, the dietary antigen ovalbumin (OVA) and the inhalant allergen Bet v 1 (the major birch pollen allergen), chemically coupled to recombinant CTB were tested for their potential to influence Th2-like immune responses. Intranasal administration of OVA-CTB prior to sensitization with OVA led to a significant decrease of antigen-specific IgE antibody levels, but a marked increase of OVA-specific IgG2a antibodies as compared to non-pretreated, sensitized animals. Antigen-specific lympho proliferative responses in vitro were reduced by 65% in the pretreated group; IL 5 and IL-4, but not IFN-gamma, production were markedly decreased in responder cells of lungs and spleens of nasally pretreated mice. In contrast, mucosal administration of rBet v 1-CTB conjugates prior to sensitization led to an up regulation of allergen-specific IgE, IgG1 and IgG2a, increased in vitro lympho proliferative responses as well as augmented production of IL-5, IL-4, IL-10 and IFN-gamma. Intranasal administration prior to sensitization of unconjugated allergens showed also contrasting effects: OVA could not significantly influence antigen-specific antibody or cytokine production, whereas intranasal pretreatment with unconjugated Bet v 1 suppressed allergen-specific immune responses in vivo and in vitro. These results demonstrated that the two antigens--in conjugated as in unconjugated form--had different effects on the Th2 immune responses. We therefore conclude that the tolerogenic or immunogenic properties of CTB--and probably also other antigen-delivery systems--strongly depend on the nature of the coupled antigen-allergen. PMID- 10383948 TI - Target cells for an immunosuppressive cytokine, glycosylation-inhibiting factor. AB - Receptors for bioactive glycosylation-inhibiting factor (GIF) were demonstrated using a bioactive mutant of recombinant human (rh) GIF, which is comparable to the suppressor T (Ts) cell-derived bioactive GIF in its affinity for the receptors on helper T (Th) hybridoma cells. Both naive T and B cells in normal mouse spleen lacked GIF receptors. However, presentation of specific antigen to naive T cells resulted in the expression of the receptors on activated T cells. Furthermore, activation of small resting B cells with F(ab')2 fragments of anti mouse IgM plus IL-4, lipopolysaccharide (LPS) plus IL-4 or LPS plus dextran sulfate induced the expression of the receptors within 48 h of B cell stimulation. It was also found that NK T cells freshly isolated from mouse spleen, but not conventional NK cells, expressed receptors for GIF. CD4(+) and CD4(-) subpopulations of NK T cells showed a similar binding capability. Mature dendritic cells derived from bone marrow did not bear the receptors. The dissociation constant (Kd) of the interaction between the bioactive rhGIF mutant and the high-affinity receptors was 10-100 pM, whereas inactive wild-type rhGIF failed to bind to the receptors. A bioactive derivative of rhGIF suppressed both IgG1 and IgE synthesis by purified B cells activated by LPS and IL-4, indicating that the binding of bioactive GIF to its receptors on activated B cells results in suppression of their differentiation. PMID- 10383949 TI - Molecular and cellular basis of the altered immune response against arsonate in irradiated A/J mice autologously reconstituted. AB - The humoral immune response to arsonate (Ars) in normal A/J mice is dominated in the late primary and particularly in the secondary response by a recurrent and dominant idiotype (CRIA) which is encoded by a single canonical combination of the variable gene segments: VHidcr11-DFL16.1-JH2 and Vkappa10-Jkappa1. Accumulation of somatic mutations within cells expressing this canonical combination or some less frequent Ig rearrangements results in the generation of high-affinity antibodies. By contrast, in partially shielded and irradiated A/J mice (autologous reconstitution) immunized with Ars-keyhole limpet hemocyanin (KLH), both the dominance of the CRIA idiotype and the affinity maturation are lost, whereas the anti-Ars antibody titer is not affected. To understand these alterations, we have analyzed a collection of 27 different anti-Ars hybridomas from nine partially shielded and irradiated A/J mice that had been immunized twice with Ars-KLH. Sequence analysis of the productively rearranged heavy chain variable region genes from those hybridomas revealed that (i) the canonical V(D)J combination was rare, (ii) the pattern of V(D)J gene usage rather corresponded to a primary repertoire with multiple gene combinations and (iii) the frequency of somatic mutations was low when compared to a normal secondary response to Ars. In addition, immunohistological analysis has shown a delay of 2 weeks in the appearance of full blown splenic germinal centers in autoreconstituting mice, as compared to controls. Such a model could be useful to understand the immunological defects found in patients transplanted with bone marrow. PMID- 10383950 TI - Differential expression of B7 co-stimulatory molecules by astrocytes correlates with T cell activation and cytokine production. AB - Whether astrocytes utilize B7:CD28 co-stimulation to activate T cells mediating CNS inflammatory disease is controversial. In this report, primary astrocytes and murine astrocyte lines, generated by immortalization at two different times, day 7 or 45 of culture, were examined for their capability to express B7 co stimulatory molecules and to participate in B7:CD28 co-stimulation. Following exposure to IFN-gamma, primary astrocytes and astrocyte lines up-regulated MHC class II and B7-2 (CD86) molecules. However, B7-1 (CD80) expression was not inducible on primary astrocytes examined after IFN-gamma stimulation beginning on day 7 or on astrocyte lines immortalized on day 7. B7-1 expression was inducible on primary astrocytes examined later and could be up-regulated on astrocyte lines immortalized later. Unlike B7-1, temporal discordant expression of other co stimulatory/adhesion molecules was not observed. Both B7-1(-)/B7-2(+) and B7 1(+)/B7-2(+) astrocyte lines were capable of stimulating proliferation of encephalitogenic Th1 cells, utilizing B7-2 for B7:CD28 co-stimulation. However, lines derived from immortalization later (B7-1(+)/B7-2(+)) were more effective in stimulating proliferation of naive myelin basic protein-specific CD4(+) T cells. Astrocyte lines that expressed both B7-1 and B7-2 also stimulated Thp cells to secrete proinflammatory Th1 cytokines, whereas lines that expressed B7-2 only stimulated Thp cells to produce a Th2 cytokine pattern. Thus, we demonstrate for the first time that individual astrocytes can differentially express B7-1 molecules, which may correlate with their ability to stimulate proinflammatory and regulatory patterns of cytokine production. These results suggest that astrocytes have potential for both promoting and down-regulating T cell responses, and that temporal differences in expression of B7 molecules should be considered when evaluating immune regulation by astrocytes. PMID- 10383951 TI - How photosynthetic bacteria harvest solar energy. PMID- 10383952 TI - Characterization of a dam mutant of Serratia marcescens and nucleotide sequence of the dam region. AB - The DNA of Serratia marcescens has N6-adenine methylation in GATC sequences. Among 2-aminopurine-sensitive mutants isolated from S. marcescens Sr41, one was identified which lacked GATC methylation. The mutant showed up to 30-fold increased spontaneous mutability and enhanced mutability after treatment with 2 aminopurine, ethyl methanesulfonate, or UV light. The gene (dam) coding for the adenine methyltransferase (Dam enzyme) of S. marcescens was identified on a gene bank plasmid which alleviated the 2-aminopurine sensitivity and the higher mutability of a dam-13::Tn9 mutant of Escherichia coli. Nucleotide sequencing revealed that the deduced amino acid sequence of Dam (270 amino acids; molecular mass, 31.3 kDa) has 72% identity to the Dam enzyme of E. coli. The dam gene is located between flanking genes which are similar to those found to the sides of the E. coli dam gene. The results of complementation studies indicated that like Dam of E. coli and unlike Dam of Vibrio cholerae, the Dam enzyme of S. marcescens plays an important role in mutation avoidance by allowing the mismatch repair enzymes to discriminate between the parental and newly synthesized strands during correction of replication errors. PMID- 10383953 TI - Amino acid residues in the omega-minus region participate in cellular localization of yeast glycosylphosphatidylinositol-attached proteins. AB - The final destination of glycosylphosphatidylinositol (GPI)-attached proteins in Saccharomyces cerevisiae is the plasma membrane or the cell wall. Two kinds of signals have been proposed for their cellular localization: (i) the specific amino acid residues V, I, or L at the site 4 or 5 amino acids upstream of the GPI attachment site (the omega site) and Y or N at the site 2 amino acids upstream of the omega site for cell wall localization and (ii) dibasic residues in the region upstream of the omega site (the omega-minus region) for plasma membrane localization. The relationships between these amino acid residues and efficiencies of cell wall incorporation were examined by constructing fusion reporter proteins from open reading frames encoding putative GPI-attached proteins. The levels of incorporation were high in the constructs containing the specific amino acid residues and quite low in those containing two basic amino acid residues in the omega-minus region. With constructs that contained neither specific residues nor two basic residues, levels of incorporation were moderate. These correlations clearly suggest that GPI-attached proteins have two different signals which act positively or negatively in cell wall incorporation for their cellular localization. PMID- 10383954 TI - Effect of rpoS mutation on the stress response and expression of virulence factors in Pseudomonas aeruginosa. AB - The sigma factor RpoS (sigmaS) has been described as a general stress response regulator that controls the expression of genes which confer increased resistance to various stresses in some gram-negative bacteria. To elucidate the role of RpoS in Pseudomonas aeruginosa physiology and pathogenesis, we constructed rpoS mutants in several strains of P. aeruginosa, including PAO1. The PAO1 rpoS mutant was subjected to various environmental stresses, and we compared the resistance phenotype of the mutant to that of the parent. The PAO1 rpoS mutant was slightly more sensitive to carbon starvation than the wild-type strain, but this phenotype was obvious only when the cells were grown in a medium supplemented with glucose as the sole carbon source. In addition, the PAO1 rpoS mutant was hypersensitive to heat shock at 50 degrees C, increased osmolarity, and prolonged exposure to high concentrations of H2O2. In accordance with the hypersensitivity to H2O2, catalase production was 60% lower in the rpoS mutant than in the parent strain. We also assessed the role of RpoS in the production of several exoproducts known to be important for virulence of P. aeruginosa. The rpoS mutant produced 50% less exotoxin A, but it produced only slightly smaller amounts of elastase and LasA protease than the parent strain. The levels of phospholipase C and casein degrading proteases were unaffected by a mutation in rpoS in PAO1. The rpoS mutation resulted in the increased production of the phenazine antibiotic pyocyanin and the siderophore pyoverdine. This increased pyocyanin production may be responsible for the enhanced virulence of the PAO1 rpoS mutant that was observed in a rat chronic-lung-infection model. In addition, the rpoS mutant displayed an altered twitching-motility phenotype, suggesting that the colonization factors, type IV fimbriae, were affected. Finally, in an alginate overproducing cystic fibrosis (CF) isolate, FRD1, the rpoS101::aacCI mutation almost completely abolished the production of alginate when the bacterium was grown in a liquid medium. On a solid medium, the FRD1 rpoS mutant produced approximately 70% less alginate than did the wild-type strain. Thus, our data indicate that although some of the functions of RpoS in P. aeruginosa physiology are similar to RpoS functions in other gram-negative bacteria, it also has some functions unique to this bacterium. PMID- 10383955 TI - Characterization of the major superoxide dismutase of Staphylococcus aureus and its role in starvation survival, stress resistance, and pathogenicity. AB - A Staphylococcus aureus mutant (SPW1) which is unable to survive long-term starvation was shown to have a transposon insertion within a gene homologous to the sodA family of manganese-dependent superoxide dismutases (SOD). Whole-cell lysates of the parental 8325-4 strain demonstrated three zones of SOD activity by nondenaturing gel electrophoresis. The activities of two of these zones were dependent on manganese for activity and were absent in SPW1. The levels of SOD activity and sodA expression were growth-phase dependent, occurring most during postexponential phase. This response was also dependent on the level of aeration of the culture, with highest activity and expression occurring only under high aeration. Expression of sodA and, consequently, SOD activity could be induced by methyl viologen but only during the transition from exponential- to postexponential-phase growth. SPW1 was less able to survive amino acid limitation and acid stress but showed no alteration in pathogenicity in a mouse abscess model of infection compared to the parental strain 8325-4. PMID- 10383957 TI - The exopolygalacturonate lyase PelW and the oligogalacturonate lyase Ogl, two cytoplasmic enzymes of pectin catabolism in Erwinia chrysanthemi 3937. AB - Erwinia chrysanthemi 3937 secretes into the external medium several pectinolytic enzymes, among which are eight isoenzymes of the endo-cleaving pectate lyases: PelA, PelB, PelC, PelD, and PelE (family 1); PelI (family 4); PelL (family 3); and PelZ (family 5). In addition, one exo-cleaving pectate lyase, PelX (family 3), has been found in the periplasm of E. chrysanthemi. The E. chrysanthemi 3937 gene kdgC has been shown to exhibit a high degree of similarity to the genes pelY of Yersinia pseudotuberculosis and pelB of Erwinia carotovora, which encode family 2 pectate lyases. However, no pectinolytic activity has been assigned to the KdgC protein. After verification of the corresponding nucleotide sequence, we cloned a longer DNA fragment and showed that this gene encodes a 553-amino-acid protein exhibiting an exo-cleaving pectate lyase activity. Thus, the kdgC gene was renamed pelW. PelW catalyzes the formation of unsaturated digalacturonates from polygalacturonate or short oligogalacturonates. PelW is located in the bacterial cytoplasm. In this compartment, PelW action could complete the degradation of pectic oligomers that was initiated by the extracellular or periplasmic pectinases and precede the action of the cytoplasmic oligogalacturonate lyase, Ogl. Both cytoplasmic pectinases, PelW and Ogl, seem to act in sequence during oligogalacturonate depolymerization, since oligomers longer than dimers are very poor substrates for Ogl but are good substrates for PelW. The estimated number of binding subsites for PelW is three, extending from subsite -2 to +1, while it is probably two for Ogl, extending from subsite -1 to +1. The activities of the two cytoplasmic lyases, PelW and Ogl, are dependent on the presence of divalent cations, since both enzymes are inhibited by EDTA. In contrast to the extracellular pectate lyases, Ca2+ is unable to restore the activity of PelW or Ogl, while several other cations, including Co2+, Mn2+, and Ni2+, can activate both cytoplasmic lyases. PMID- 10383956 TI - Effects of ethyl and benzyl analogues of spermine on Escherichia coli peptidyltransferase activity, polyamine transport, and cellular growth. AB - Various ethyl and benzyl spermine analogues, including the anticancer agent N1,N12-bis(ethyl)spermine, were studied for their ability to affect the growth of cultured Escherichia coli cells, to inhibit [3H]putrescine and [3H]spermine uptake into cells, and to modulate the peptidyltransferase activity (EC 2. 3. 2. 12). Relative to other cell lines, growth of E. coli was uniquely insensitive to these analogues. Nevertheless, these analogues conferred similar modulation of in vitro protein synthesis and inhibition of [3H]putrescine and [3H]spermine uptake, as is seen in other cell types. Thus, both ethyl and benzyl analogues of spermine not only promote the formation and stabilization of the initiator ribosomal ternary complex, but they also have a sparing effect on the Mg2+ requirements. Also, in a complete cell-free protein-synthesizing system, these analogues at low concentrations stimulated peptide bond formation, whereas at higher concentrations, they inhibited the reaction. The ranking order for stimulation of peptide-bond formation by the analogues was N4,N9-dibenzylspermine > N4, N9 bis(ethyl)spermine congruent with N1-ethylspermine > N1, N12-bis(ethyl)spermine, whereas the order of analogue potency regarding the inhibitory effect was inverted, with inhibition constant values of 10, 3.1, 1.5, and 0.98 microM, respectively. Although the above analogues failed to interact with the putrescine specific uptake system, they exhibited high affinity for the polyamine uptake system encoded by the potABCD operon. Despite this fact, none of the analogues could be internalized by the polyamine transport system, and therefore they could not influence the intracellular polyamine pools and growth of E. coli cells. PMID- 10383958 TI - Coordinate transcriptional control in the hyperthermophilic archaeon Sulfolobus solfataricus. AB - The existence of a global gene regulatory system in the hyperthermophilic archaeon Sulfolobus solfataricus is described. The system is responsive to carbon source quality and acts at the level of transcription to coordinate synthesis of three physically unlinked glycosyl hydrolases implicated in carbohydrate utilization. The specific activities of three enzymes, an alpha-glucosidase (malA), a beta-glycosidase (lacS), and an alpha-amylase, were reduced 4-, 20-, and 10-fold, respectively, in response to the addition of supplementary carbon sources to a minimal sucrose medium. Western blot analysis using anti-alpha glucosidase and anti-beta-glycosidase antibodies indicated that reduced enzyme activities resulted exclusively from decreased enzyme levels. Northern blot analysis of malA and lacS mRNAs revealed that changes in enzyme abundance arose primarily from reductions in transcript concentrations. Culture conditions precipitating rapid changes in lacS gene expression were established to determine the response time of the regulatory system in vivo. Full induction occurred within a single generation whereas full repression occurred more slowly, requiring nearly 38 generations. Since lacS mRNA abundance changed much more rapidly in response to a nutrient down shift than to a nutrient up shift, transcript synthesis rather than degradation likely plays a role in the regulatory response. PMID- 10383959 TI - Elements involved in catabolite repression and substrate induction of the lactose operon in Lactobacillus casei. AB - In Lactobacillus casei ATCC 393, the chromosomally encoded lactose operon, lacTEGF, encodes an antiterminator protein (LacT), lactose-specific phosphoenolpyruvate-dependent phosphotransferase system (PTS) elements (LacE and LacF), and a phospho-beta-galactosidase. lacT, lacE, and lacF mutant strains were constructed by double crossover. The lacT strain displayed constitutive termination at a ribonucleic antiterminator (RAT) site, whereas lacE and lacF mutants showed an inducer-independent antiterminator activity, as shown analysis of enzyme activity obtained from transcriptional fusions of lac promoter (lacp) and lacpDeltaRAT with the Escherichia coli gusA gene in the different lac mutants. These results strongly suggest that in vivo under noninducing conditions, the lactose-specific PTS elements negatively modulate LacT activity. Northern blot analysis detected a 100-nucleotide transcript starting at the transcription start site and ending a consensus RAT sequence and terminator region. In a ccpA mutant, transcription initiation was derepressed but no elongation through the terminator was observed in the presence of glucose and the inducing sugar, lactose. Full expression of lacTEGF was found only in a man ccpA double mutant, indicating that PTS elements are involved in the CcpA-independent catabolite repression mechanism probably via LacT. PMID- 10383960 TI - The "green" form I ribulose 1,5-bisphosphate carboxylase/oxygenase from the nonsulfur purple bacterium Rhodobacter capsulatus. AB - Form I ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) of the Calvin Benson-Bassham cycle may be divided into two broad phylogenetic groups, referred to as red-like and green-like, based on deduced large subunit amino acid sequences. Unlike the form I enzyme from the closely related organism Rhodobacter sphaeroides, the form I RubisCO from R. capsulatus is a member of the green-like group and closely resembles the enzyme from certain chemoautotrophic proteobacteria and cyanobacteria. As the enzymatic properties of this type of RubisCO have not been well studied in a system that offers facile genetic manipulation, we purified the R. capsulatus form I enzyme and determined its basic kinetic properties. The enzyme exhibited an extremely low substrate specificity factor, which is congruent with its previously determined sequence similarity to form I enzymes from chemoautotrophs and cyanobacteria. The enzymological results reported here are thus strongly supportive of the previously suggested horizontal gene transfer that most likely occurred between a green-like RubisCO-containing bacterium and a predecessor to R. capsulatus. Expression results from hybrid and chimeric enzyme plasmid constructs, made with large and small subunit genes from R. capsulatus and R. sphaeroides, also supported the unrelatedness of these two enzymes and were consistent with the recently proposed phylogenetic placement of R. capsulatus form I RubisCO. The R. capsulatus form I enzyme was found to be subject to a time-dependent fallover in activity and possessed a high affinity for CO2, unlike the closely similar cyanobacterial RubisCO, which does not exhibit fallover and possesses an extremely low affinity for CO2. These latter results suggest definite approaches to elucidate the molecular basis for fallover and CO2 affinity. PMID- 10383961 TI - Expression of the sigmaB-dependent general stress regulon confers multiple stress resistance in Bacillus subtilis. AB - The alternative sigma factor sigmaB of Bacillus subtilis is required for the induction of approximately 100 genes after the imposition of a whole range of stresses and energy limitation. In this study, we investigated the impact of a null mutation in sigB on the stress and starvation survival of B. subtilis. sigB mutants which failed to induce the regulon following stress displayed an at least 50- to 100-fold decrease in survival of severe heat (54 degrees C) or ethanol (9%) shock, salt (10%) stress, and acid (pH 4.3) stress, as well as freezing and desiccation, compared to the wild type. Preloading cells with sigmaB-dependent general stress proteins prior to growth-inhibiting stress conferred considerable protection against heat and salt. Exhaustion of glucose or phosphate induced the sigmaB response, but surprisingly, sigmaB did not seem to be required for starvation survival. Starved wild-type cells exhibited about 10-fold greater resistance to salt stress than exponentially growing cells. The data argue that the expression of sigmaB-dependent genes provides nonsporulated B. subtilis cells with a nonspecific multiple stress resistance that may be relevant for stress survival in the natural ecosystem. PMID- 10383962 TI - Interaction of Azospirillum lipoferum with wheat germ agglutinin stimulates nitrogen fixation. AB - In vitro, the nitrogen fixation capability of A. lipoferum is efficiently increased in the presence of wheat germ agglutinin (WGA). A putative WGA-binding receptor, a 32-kDa protein, was detected in the cell capsule. The stimulatory effect required N-acetyl-D-glucosamine dimer (GlcNAcdi) terminated sugar side chains of the receptor and was dependent on the number of GlcNAcdi links involved in receptor-WGA interface. Binding to the primary sugar binding sites on WGA had a larger stimulatory effect than binding to the secondary sites. The WGA-receptor complex generated stimulus led to elevated transcription of the nifH and nifA genes and of the glnBA gene cluster but not of the glnA gene from its own promoter. There may well be a signalling cascade contributing to the regulation of nitrogen fixation. PMID- 10383963 TI - Analysis of peptidoglycan structure from vegetative cells of Bacillus subtilis 168 and role of PBP 5 in peptidoglycan maturation. AB - The composition and fine structure of the vegetative cell wall peptidoglycan from Bacillus subtilis were determined by analysis of its constituent muropeptides. The structures of 39 muropeptides, representing 97% of the total peptidoglycan, were elucidated. About 99% analyzed muropeptides in B. subtilis vegetative cell peptidoglycan have the free carboxylic group of diaminopimelic acid amidated. Anhydromuropeptides and products missing a glucosamine at the nonreducing terminus account for 0.4 and 1.5%, respectively, of the total muropeptides. These two types of muropeptides are suggested to end glycan strands. An unexpected feature of B. subtilis muropeptides was the occurrence of a glycine residue in position 5 of the peptide side chain on monomers or oligomers, which account for 2.7% of the total muropeptides. This amount is, however, dependent on the composition of the growth media. Potential attachment sites for anionic polymers to peptidoglycan occur on dominant muropeptides and account for 2.1% of the total. B. subtilis peptidoglycan is incompletely digested by lysozyme due to de-N acetylation of glucosamine, which occurs on 17.3% of muropeptides. The cross linking index of the polymer changes with the growth phase. It is highest in late stationary phase, with a value of 33.2 or 44% per muramic acid residue, as determined by reverse-phase high-pressure liquid chromatography or gel filtration, respectively. Analysis of the muropeptide composition of a dacA (PBP 5) mutant shows a dramatic decrease of muropeptides with tripeptide side chains and an increase or appearance of muropeptides with pentapeptide side chains in monomers or oligomers. The total muropeptides with pentapeptide side chains accounts for almost 82% in the dacA mutant. This major low-molecular-weight PBP (DD-carboxypeptidase) is suggested to play a role in peptidoglycan maturation. PMID- 10383964 TI - Regulation of podJ expression during the Caulobacter crescentus cell cycle. AB - The polar organelle development gene, podJ, is expressed during the swarmer-to stalked cell transition of the Caulobacter crescentus cell cycle. Mutants with insertions that inactivate the podJ gene are nonchemotactic, deficient in rosette formation, and resistant to polar bacteriophage, but they divide normally. In contrast, hyperexpression of podJ results in a lethal cell division defect. Nucleotide sequence analysis of the podJ promoter region revealed a binding site for the global response regulator, CtrA. Deletion of this site results in increased overall promoter activity, suggesting that CtrA is a negative regulator of the podJ promoter. Furthermore, synchronization studies have indicated that temporal regulation is not dependent on the presence of the CtrA binding site. Thus, although the level of podJ promoter activity is dependent on the CtrA binding site, the temporal control of podJ promoter expression is dependent on other factors. PMID- 10383965 TI - Ribonucleotide reduction in Pseudomonas species: simultaneous presence of active enzymes from different classes. AB - Three separate classes of ribonucleotide reductases exist in nature. They differ widely in protein structure. Class I enzymes are found in aerobic bacteria and eukaryotes; class II enzymes are found in aerobic and anaerobic bacteria; class III enzymes are found in strict and facultative anaerobic bacteria. Usually, but not always, one organism contains only one or two (in facultative anaerobes) classes. Surprisingly, the genomic sequence of Pseudomonas aeruginosa contains sequences for each of the three classes. Here, we show by DNA hybridization that other species of Pseudomonas also contain the genes for three classes. Extracts from P. aeruginosa and P. stutzeri grown aerobically or microaerobically contain active class I and II enzymes, whereas we could not demonstrate class III activity. Unexpectedly, class I activity increased greatly during microaerobic conditions. The enzymes were separated, and the large proteins of the class I enzymes were obtained in close to homogeneous form. The catalytic properties of all enzymes are similar to those of other bacterial reductases. However, the Pseudomonas class I reductases required the continuous presence of oxygen during catalysis, unlike the corresponding Escherichia coli enzyme but similar to the mouse enzyme. In similarity searches, the amino acid sequence of the class I enzyme of P. aeruginosa was more related to that of eukaryotes than to that of E. coli or other proteobacteria, with the large protein showing 42% identity to that of the mouse, suggesting the possibility of a horizontal transfer of the gene. The results raise many questions concerning the physiological function and evolution of the three classes in Pseudomonas species. PMID- 10383967 TI - Type 2 NADH dehydrogenases in the cyanobacterium Synechocystis sp. strain PCC 6803 are involved in regulation rather than respiration. AB - Analysis of the genome of Synechocystis sp. strain PCC 6803 reveals three open reading frames (slr0851, slr1743, and sll1484) that may code for type 2 NAD(P)H dehydrogenases (NDH-2). The sequence similarity between the translated open reading frames and NDH-2s from other organisms is low, generally not exceeding 30% identity. However, NAD(P)H and flavin adenine dinucleotide binding motifs are conserved in all three putative NDH-2s in Synechocystis sp. strain PCC 6803. The three open reading frames were cloned, and deletion constructs were made for each. An expression construct containing one of the three open reading frames, slr1743, was able to functionally complement an Escherichia coli mutant lacking both NDH-1s and NDH-2s. Therefore, slr0851, slr1743, and sll1484 have been designated ndbA, ndbB, and ndbC, respectively. Strains that lacked one or more of the ndb genes were created in wild-type and photosystem (PS) I-less backgrounds. Deletion of ndb genes led to small changes in photoautotrophic growth rates and respiratory activities. Electron transfer rates into the plastoquinone pool in thylakoids in darkness were consistent with the presence of a small amount of NDH 2 activity in thylakoids. No difference was observed between wild-type and the Ndb-less strains in the banding patterns seen on native gels when stained for either NADH or NADPH dehydrogenase activity, indicating that the Ndb proteins do not accumulate to high levels. A striking phenotype of the PS I-less background strains lacking one or more of the NDH-2s is that they were able to grow at high light intensities that were lethal to the control strain but they retained normal PS II activity. We suggest that the Ndb proteins in Synechocystis sp. strain PCC 6803 are redox sensors and that they play a regulatory role responding to the redox state of the plastoquinone pool. PMID- 10383966 TI - Escherichia coli mutants lacking all possible combinations of eight penicillin binding proteins: viability, characteristics, and implications for peptidoglycan synthesis. AB - The penicillin binding proteins (PBPs) synthesize and remodel peptidoglycan, the structural component of the bacterial cell wall. Much is known about the biochemistry of these proteins, but little is known about their biological roles. To better understand the contributions these proteins make to the physiology of Escherichia coli, we constructed 192 mutants from which eight PBP genes were deleted in every possible combination. The genes encoding PBPs 1a, 1b, 4, 5, 6, and 7, AmpC, and AmpH were cloned, and from each gene an internal coding sequence was removed and replaced with a kanamycin resistance cassette flanked by two res sites from plasmid RP4. Deletion of individual genes was accomplished by transferring each interrupted gene onto the chromosome of E. coli via lambda phage transduction and selecting for kanamycin-resistant recombinants. Afterwards, the kanamycin resistance cassette was removed from each mutant strain by supplying ParA resolvase in trans, yielding a strain in which a long segment of the original PBP gene was deleted and replaced by an 8-bp res site. These kanamycin-sensitive mutants were used as recipients in further rounds of replacement mutagenesis, resulting in a set of strains lacking from one to seven PBPs. In addition, the dacD gene was deleted from two septuple mutants, creating strains lacking eight genes. The only deletion combinations not produced were those lacking both PBPs 1a and 1b because such a combination is lethal. Surprisingly, all other deletion mutants were viable even though, at the extreme, 8 of the 12 known PBPs had been eliminated. Furthermore, when both PBPs 2 and 3 were inactivated by the beta-lactams mecillinam and aztreonam, respectively, several mutants did not lyse but continued to grow as enlarged spheres, so that one mutant synthesized osmotically resistant peptidoglycan when only 2 of 12 PBPs (PBPs 1b and 1c) remained active. These results have important implications for current models of peptidoglycan biosynthesis, for understanding the evolution of the bacterial sacculus, and for interpreting results derived by mutating unknown open reading frames in genome projects. In addition, members of the set of PBP mutants will provide excellent starting points for answering fundamental questions about other aspects of cell wall metabolism. PMID- 10383968 TI - Membrane association and multimerization of secreton component pulC. AB - The PulC component of the Klebsiella oxytoca pullulanase secretion machinery (the secreton) was found by subcellular fractionation to be associated with both the cytoplasmic (inner) and outer membranes. Association with the outer membrane was independent of other secreton components, including the outer membrane protein PulD (secretin). The association of PulC with the inner membrane is mediated by the signal anchor sequence located close to its N terminus. These results suggest that PulC forms a bridge between the two membranes that is disrupted when bacteria are broken open for fractionation. Neither the signal anchor sequence nor the cytoplasmic N-terminal region that precedes it was found to be required for PulC function, indicating that PulC does not undergo sequence-specific interactions with other cytoplasmic membrane proteins. Cross-linking of whole cells resulted in the formation of a ca. 110-kDa band that reacted with PulC specific serum and whose detection depended on the presence of PulD. However, antibodies against PulD failed to react with this band, suggesting that it could be a homo-PulC trimer whose formation requires PulD. The data are discussed in terms of the possible role of PulC in energy transduction for exoprotein secretion. PMID- 10383969 TI - Structure-function analysis of XcpP, a component involved in general secretory pathway-dependent protein secretion in Pseudomonas aeruginosa. AB - The general secretory pathway of Pseudomonas aeruginosa is required for the transport of signal peptide-containing exoproteins across the cell envelope. After completion of the Sec-dependent translocation of exoproteins across the inner membrane and cleavage of the signal peptide, the Xcp machinery mediates translocation across the outer membrane. This machinery consists of 12 components, of which XcpQ (GspD) is the sole outer membrane protein. XcpQ forms a multimeric ring-shaped structure, with a central opening through which exoproteins could pass to reach the medium. Surprisingly, all of the other Xcp proteins are located in or are associated with the cytoplasmic membrane. This study is focused on the characteristics of one such cytoplasmic membrane protein, XcpP. An xcpP mutant demonstrated that the product of this gene is indeed an essential element of the P. aeruginosa secretion machinery. Construction and analysis of truncated forms of XcpP made it possible to define essential domains for the function of the protein. Some of these domains, such as the N-terminal transmembrane domain and a coiled-coil structure identified at the C terminus of XcpP, may be involved in protein-protein interaction during the assembly of the secretory apparatus. PMID- 10383970 TI - Temperature-sensitive motility of Sulfolobus acidocaldarius influences population distribution in extreme environments. AB - A three-dimensional tracking microscope was used to quantify the effects of temperature (50 to 80 degrees C) and pH (2 to 4) on the motility of Sulfolobus acidocaldarius, a thermoacidophilic archaeon. Swimming speed and run time increased with temperature but remained relatively unchanged with increasing pH. These results were consistent with reported changes in the rate of respiration of S. acidocaldarius as a function of temperature and pH. Cells exhibited a forward biased turn angle distribution with a mean of 54 degrees. Cell trajectories during a run were in the shape of right-handed helices. A cellular dynamics simulation was used to test the hypothesis that a population of S. acidocaldarius cells could distribute preferentially in a spatial temperature gradient due to variation in swimming speed. Simulation results showed that a population of cells could migrate from a higher to a lower temperature in the presence of sharp temperature gradients. This simulation result was achieved without incorporating the ability of cells to sense a temporal thermal gradient; thus, the response was not thermotactic. We postulate that this temperature-sensitive motility is one survival mechanism of S. acidocaldarius that allows this organism to move away from lethal hot spots in its hydrothermal environment. PMID- 10383971 TI - Characterization of the Moraxella catarrhalis uspA1 and uspA2 genes and their encoded products. AB - The uspA1 and uspA2 genes of M. catarrhalis O35E encode two different surface exposed proteins which were previously shown to share a 140-amino-acid region with 93% identity (C. Aebi, I. Maciver, J. L. Latimer, L. D. Cope, M. K. Stevens, S. E. Thomas, G. H. McCracken, Jr., and E. J. Hansen, Infect. Immun. 65:4367 4377, 1997). The N-terminal amino acid sequences of the mature forms of both UspA1 and UspA2 from strain O35E were determined after enzymatic treatment to remove the N-terminal pyroglutamyl residue that had blocked Edman degradation. Mass spectrometric analysis indicated that the molecular mass of UspA1 from M. catarrhalis O35E was 83,500 +/- 116 Da. Nucleotide sequence analysis of the uspA1 and uspA2 genes from three other M. catarrhalis strains (TTA24, ATCC 25238, and V1171) revealed that the encoded protein products were very similar to those from strain O35E. Western blot analysis was used to confirm that each of these three strains of M. catarrhalis expressed both UspA1 and UspA2 proteins. Several different and repetitive amino acid motifs were present in both UspA1 and UspA2 from these four strains, and some of these were predicted to form coiled coils. Linear DNA templates were used in an in vitro transcription-translation system to determine the sizes of the monomeric forms of the UspA1 and UspA2 proteins from strains O35E and TTA24. PMID- 10383972 TI - The extracellular xylan degradative system in Clostridium cellulolyticum cultivated on xylan: evidence for cell-free cellulosome production. AB - In this study, we demonstrate that the cellulosome of Clostridium cellulolyticum grown on xylan is not associated with the bacterial cell. Indeed, the large majority of the activity (about 90%) is localized in the cell-free fraction when the bacterium is grown on xylan. Furthermore, about 70% of the detected xylanase activity is associated with cell-free high-molecular-weight complexes containing avicelase activity and the cellulosomal scaffolding protein CipC. The same repartition is observed with carboxymethyl cellulase activity. The cellulose adhesion of xylan-grown cells is sharply reduced in comparison with cellulose grown cells. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed that cellulosomes derived from xylan- and cellulose-grown cells have different compositions. In both cases, the scaffolding protein CipC is present, but the relative proportions of the other components is dramatically changed depending on the growth substrate. We propose that, depending on the growth substrate, C. cellulolyticum is able to regulate the cell association and cellulose adhesion of cellulosomes and regulate cellulosomal composition. PMID- 10383973 TI - Specific chromosome alterations in fluconazole-resistant mutants of Candida albicans. AB - The exposure of Candida albicans to fluconazole resulted in the nondisjunction of two specific chromosomes in 17 drug-resistant mutants, each obtained by an independent mutational event. The chromosomal changes occurred at high frequencies and were related to the duration of the drug exposure. The loss of one homologue of chromosome 4 occurred after incubation on a fluconazole medium for 7 days. A second change, the gain of one copy of chromosome 3, was observed after exposure for 35 or 40 days. We found that the mRNA levels of ERG11, CDR1, CDR2, and MDR1, the candidate fluconazole resistance genes, remained either the same or were diminished. The lack of overexpression of putative drug pumps or the drug target indicated that some other mechanism(s) may be operating. The fluconazole resistance phenotype, electrophoretic karyotypes, and transcript levels of mutants were stable after growth for 112 generations in the absence of fluconazole. This is the first report to demonstrate that resistance to fluconazole can be dependent on chromosomal nondisjunction. Furthermore, we suggest that a low-level resistance to fluconazole arising during the early stages of clinical treatment may occur by this mechanism. These results support our earlier hypothesis that changes in C. albicans chromosome number is a common means to control a resource of potentially beneficial genes that are related to important cellular functions. PMID- 10383975 TI - Site-specific recombination of temperate Myxococcus xanthus phage Mx8: regulation of integrase activity by reversible, covalent modification. AB - Temperate Myxococcus xanthus phage Mx8 integrates into the attB locus of the M. xanthus genome. The phage attachment site, attP, is required in cis for integration and lies within the int (integrase) coding sequence. Site-specific integration of Mx8 alters the 3' end of int to generate the modified intX gene, which encodes a less active form of integrase with a different C terminus. The phage-encoded (Int) form of integrase promotes attP x attB recombination more efficiently than attR x attB, attL x attB, or attB x attB recombination. The attP and attB sites share a common core. Sequences flanking both sides of the attP core within the int gene are necessary for attP function. This information shows that the directionality of the integration reaction depends on arm sequences flanking both sides of the attP core. Expression of the uoi gene immediately upstream of int inhibits integrative (attP x attB) recombination, supporting the idea that uoi encodes the Mx8 excisionase. Integrase catalyzes a reaction that alters the primary sequence of its gene; the change in the primary amino acid sequence of Mx8 integrase resulting from the reaction that it catalyzes is a novel mechanism by which the reversible, covalent modification of an enzyme is used to regulate its specific activity. The lower specific activity of the prophage-encoded IntX integrase acts to limit excisive site-specific recombination in lysogens carrying a single Mx8 prophage, which are less immune to superinfection than lysogens carrying multiple, tandem prophages. Thus, this mechanism serves to regulate Mx8 site-specific recombination and superinfection immunity coordinately and thereby to preserve the integrity of the lysogenic state. PMID- 10383974 TI - Site-specific recombination of temperate Myxococcus xanthus phage Mx8: genetic elements required for integration. AB - Like most temperate bacteriophages, phage Mx8 integrates into a preferred locus on the genome of its host, Myxococcus xanthus, by a mechanism of site-specific recombination. The Mx8 int-attP genes required for integration map within a 2.2 kilobase-pair (kb) fragment of the phage genome. When this fragment is subcloned into a plasmid vector, it facilitates the site-specific integration of the plasmid into the 3' ends of either of two tandem tRNAAsp genes, trnD1 and trnD2, located within the attB locus of the M. xanthus genome. Although Int-mediated site-specific recombination occurs between attP and either attB1 (within trnD1) or attB2 (within trnD2), the attP x attB1 reaction is highly favored and often is accompanied by a deletion between attB1 and attB2. The int gene is the only Mx8 gene required in trans for attP x attB recombination. The int promoter lies within the 106-bp region immediately upstream of one of two alternate GTG start codons, GTG-5208 (GTG at bp 5208) and GTG-5085, for integrase and likely is repressed in the prophage state. All but the C-terminal 30 amino acid residues of the Int protein are required for its ability to mediate attP x attB recombination efficiently. The attP core lies within the int coding sequence, and the product of integration is a prophage in which the 3' end of int is replaced by host sequences. The prophage intX gene is predicted to encode an integrase with a different C terminus. PMID- 10383976 TI - Cloning and expression of cadD, a new cadmium resistance gene of Staphylococcus aureus. AB - A cadmium resistance gene, designated cadD, has been identified in and cloned from the Staphylococcus aureus plasmid pRW001. The gene is part of a two component operon which contains the resistance gene cadD and an inactive regulatory gene, cadX*. A high degree of sequence similarity was observed between cadD and the cadB-like gene from S. lugdunensis, but no significant similarity was found with either cadA or cadB from the S. aureus plasmids pI258 and pII147. The positive regulatory gene cadX* is identical to cadX from pLUG10 over a stretch of 78 codons beginning at the N terminus, but it is truncated at this point and inactive. Sequence analysis showed that the cadmium resistance operon resides on a 3,972-bp element that is flanked by direct repeats of IS257. The expression of cadD in S. aureus and Bacillus subtilis resulted in low-level resistance to cadmium; in contrast, cadA and cadB from S. aureus induced higher level resistance. However, when the truncated version of cadX contained in pRW001 is complemented in trans with cadX from plasmid pLUG10, resistance increased approximately 10-fold suggesting that the cadmium resistance operons from pRW001 and pLUG10 are evolutionarily related. Moreover, the truncated version of cadX contained in pRW001 is nonfunctional and may have been generated by deletion during recombination to acquire the cadmium resistance element. PMID- 10383977 TI - Energy conservation by the H2:heterodisulfide oxidoreductase from Methanosarcina mazei Go1: identification of two proton-translocating segments. AB - The membrane-bound H2:heterodisulfide oxidoreductase system of the methanogenic archaeon Methanosarcina mazei Go1 catalyzed the H2-dependent reduction of 2 hydroxyphenazine and the dihydro-2-hydroxyphenazine-dependent reduction of the heterodisulfide of HS-CoM and HS-CoB (CoM-S-S-CoB). Washed inverted vesicles of this organism were found to couple both processes with the transfer of protons across the cytoplasmic membrane. The maximal H+/2e- ratio was 0.9 for each reaction. The electrochemical proton gradient (DeltamicroH+) thereby generated was shown to drive ATP synthesis from ADP plus Pi, exhibiting stoichiometries of 0.25 ATP synthesized per two electrons transported for both partial reactions. ATP synthesis and the generation of DeltamicroH+ were abolished by the uncoupler 3,5-di-tert-butyl-4-hydroxybenzylidenemalononitrile (SF 6847). The ATP synthase inhibitor N,N'-dicyclohexylcarbodiimide did not affect H+ translocation but led to an almost complete inhibition of ATP synthesis and decreased the electron transport rates. The latter effect was relieved by the addition of SF 6847. Thus, the energy-conserving systems showed a stringent coupling which resembles the phenomenon of respiratory control. The results indicate that two different proton translocating segments are present in the H2:heterodisulfide oxidoreductase system; the first involves the 2-hydroxyphenazine-dependent hydrogenase, and the second involves the heterodisulfide reductase. PMID- 10383979 TI - Nonribosomal peptide synthesis and toxigenicity of cyanobacteria. AB - Nonribosomal peptide synthesis is achieved in prokaryotes and lower eukaryotes by the thiotemplate function of large, modular enzyme complexes known collectively as peptide synthetases. These and other multifunctional enzyme complexes, such as polyketide synthases, are of interest due to their use in unnatural-product or combinatorial biosynthesis (R. McDaniel, S. Ebert-Khosla, D. A. Hopwood, and C. Khosla, Science 262:1546-1557, 1993; T. Stachelhaus, A. Schneider, and M. A. Marahiel, Science 269:69-72, 1995). Most nonribosomal peptides from microorganisms are classified as secondary metabolites; that is, they rarely have a role in primary metabolism, growth, or reproduction but have evolved to somehow benefit the producing organisms. Cyanobacteria produce a myriad array of secondary metabolites, including alkaloids, polyketides, and nonribosomal peptides, some of which are potent toxins. This paper addresses the molecular genetic basis of nonribosomal peptide synthesis in diverse species of cyanobacteria. Amplification of peptide synthetase genes was achieved by use of degenerate primers directed to conserved functional motifs of these modular enzyme complexes. Specific detection of the gene cluster encoding the biosynthetic pathway of the cyanobacterial toxin microcystin was shown for both cultured and uncultured samples. Blot hybridizations, DNA amplifications, sequencing, and evolutionary analysis revealed a broad distribution of peptide synthetase gene orthologues in cyanobacteria. The results demonstrate a molecular approach to assessing preexpression microbial functional diversity in uncultured cyanobacteria. The nonribosomal peptide biosynthetic pathways detected may lead to the discovery and engineering of novel antibiotics, immunosuppressants, or antiviral agents. PMID- 10383978 TI - sigmaK can negatively regulate sigE expression by two different mechanisms during sporulation of Bacillus subtilis. AB - Temporal and spatial gene regulation during Bacillus subtilis sporulation involves the activation and inactivation of multiple sigma subunits of RNA polymerase in a cascade. In the mother cell compartment of sporulating cells, expression of the sigE gene, encoding the earlier-acting sigma factor, sigmaE, is negatively regulated by the later-acting sigma factor, sigmaK. Here, it is shown that the negative feedback loop does not require SinR, an inhibitor of sigE transcription. Production of sigmaK about 1 h earlier than normal does affect Spo0A, which when phosphorylated is an activator of sigE transcription. A mutation in the spo0A gene, which bypasses the phosphorelay leading to the phosphorylation of Spo0A, diminished the negative effect of early sigmaK production on sigE expression early in sporulation. Also, early production of sigmaK reduced expression of other Spo0A-dependent genes but not expression of the Spo0A-independent ald gene. In contrast, both sigE and ald were overexpressed late in development of cells that fail to make sigmaK. The ald promoter, like the sigE promoter, is believed to be recognized by sigmaA RNA polymerase, suggesting that sigmaK may inhibit sigmaA activity late in sporulation. To exert this negative effect, sigmaK must be transcriptionally active. A mutant form of sigmaK that associates with core RNA polymerase, but does not direct transcription of a sigmaK-dependent gene, failed to negatively regulate expression of sigE or ald late in development. On the other hand, the negative effect of early sigmaK production on sigE expression early in sporulation did not require transcriptional activity of sigmaK RNA polymerase. These results demonstrate that sigmaK can negatively regulate sigE expression by two different mechanisms, one observed when sigmaK is produced earlier than normal, which does not require sigmaK to be transcriptionally active and affects Spo0A, and the other observed when sigmaK is produced at the normal time, which requires sigmaK RNA polymerase transcriptional activity. The latter mechanism facilitates the switch from sigmaE to sigmaK in the cascade controlling mother cell gene expression. PMID- 10383980 TI - Molecular cloning and functional expression of alternative oxidase from Candida albicans. AB - The AOX1 gene, which encodes an alternative oxidase, was isolated from the genomic DNA library of Candida albicans. The gene encodes a polypeptide consisting of 379 amino acids with a calculated molecular mass of 43,975 Da. The aox1/aox1 mutant strain did not show cyanide-resistant respiration under normal conditions but could still induce cyanide-resistant respiration when treated with antimycin A. The measurement of respiratory activity and Western blot analysis suggested the presence of another AOX. When C. albicans AOX1 was expressed in alternative oxidase-deficient Saccharomyces cerevisiae, it could confer cyanide resistant respiration on S. cerevisiae. PMID- 10383981 TI - Isolation and properties of Enterococcus hirae mutants defective in the potassium/proton antiport system. AB - A K+/H+ antiporter regulates cytoplasmic pH in Enterococcus hirae growing at alkaline pH. Mutants defective in this antiport activity were alkaline pH sensitive. One mutant, Pop1, lacked both K+/methylamine exchange at pH 9.5 and concomitant acidification of cytoplasmic pH. Pop1 grew well at pHs below 8 but did not at pHs above 9, conditions under which cytoplasmic pH was not fully acidified. PMID- 10383982 TI - Identification of a putative P-transporter operon in the genome of a Burkholderia strain living inside the arbuscular mycorrhizal fungus Gigaspora margarita. AB - This article reports the identification of a putative P-transporter operon in the genome of a Burkholderia sp. living in the cytoplasm of the arbuscular mycorrhizal fungus Gigaspora margarita. Its presence suggests that Burkholderia sp. has the potential for P uptake from this environment. This finding raises new questions concerning the importance of intracellular bacteria for mycorrhizal symbiosis. PMID- 10383983 TI - Identification and expression of the genes encoding a reactivating factor for adenosylcobalamin-dependent glycerol dehydratase. AB - Adenosylcobalamin-dependent glycerol dehydratase undergoes inactivation by glycerol, the physiological substrate, during catalysis. In permeabilized cells of Klebsiella pneumoniae, the inactivated enzyme is reactivated in the presence of ATP, Mg2+, and adenosylcobalamin. We identified the two open reading frames as the genes for a reactivating factor for glycerol dehydratase and designated them gdrA and gdrB. The reactivation of the inactivated glycerol dehydratase by the gene products was confirmed in permeabilized recombinant Escherichia coli cells coexpressing GdrA and GdrB proteins with glycerol dehydratase. PMID- 10383984 TI - ScoC regulates peptide transport and sporulation initiation in Bacillus subtilis. AB - Oligopeptides are transported into Bacillus subtilis by two ABC transport systems, App and Opp. Transcription of the operon encoding the Opp system was found to occur during exponential growth, whereas the app operon was induced at the onset of stationary phase. Transcription of both operons was completely curtailed by overproduction of the ScoC regulator from a multicopy plasmid and was enhanced in strains with the scoC locus deleted. ScoC, a member of the MarR family of transcription regulators, is known from previous studies to be a negative regulator of sporulation and of protease production that acts by binding directly to the promoters of the genes it regulates. Since peptide transport is essential for inactivation of the negative regulation of sporulation by Rap phosphatases, the control of ScoC transcription repression activity plays a crucial role in the initiation of sporulation. PMID- 10383985 TI - The pvc gene cluster of Pseudomonas aeruginosa: role in synthesis of the pyoverdine chromophore and regulation by PtxR and PvdS. AB - A putative operon of four genes implicated in the synthesis of the chromophore moiety of the Pseudomonas aeruginosa siderophore pyoverdine, dubbed pvcABCD (where pvc stands for pyoverdine chromophore), was cloned and sequenced. Mutational inactivation of the pvc genes abrogated pyoverdine biosynthesis, consistent with their involvement in the biosynthesis of this siderophore. pvcABCD expression was negatively regulated by iron and positively regulated by both PvdS, the alternate sigma factor required for pyoverdine biosynthesis, and PtxR, a LysR family activator previously implicated in exotoxin A regulation. PMID- 10383986 TI - Mutations in catabolite control protein CcpA separating growth effects from catabolite repression. AB - Carbon catabolite repression in Bacillus megaterium is mediated by the transcriptional regulator CcpA. A chromosomal deletion of ccpA eliminates catabolite repression and reduces the growth rate on glucose. We describe four single-amino-acid mutations in CcpA which separate the growth effect from catabolite repression, suggesting distinct regulatory pathways for these phenotypes. PMID- 10383987 TI - Nitric oxide is a signal for NNR-mediated transcription activation in Paracoccus denitrificans. AB - By using the 'lacZ gene, the activities of the nirI, nirS, and norC promoters were assayed in the wild type and in NNR-deficient mutants of Paracoccus denitrificans grown under various growth conditions. In addition, induction profiles of the three promoters in response to the presence of various nitrogenous oxides were determined. Transcription from the three promoters required the absence of oxygen and the presence both of the transcriptional activator NNR and of nitric oxide. The activity of the nnr promoter itself was halved after the cells had been switched from aerobic respiration to denitrification. This response was apparently not a result of autoregulation or of regulation by FnrP, since the nnr promoter was as active in the wild-type strain as it was in NNR- or FnrP-deficient mutants. PMID- 10383988 TI - Swarming by Pseudomonas syringae B728a requires gacS (lemA) and gacA but not the acyl-homoserine lactone biosynthetic gene ahlI. AB - Pseudomonas syringae pv. syringae B728a, a causal agent of bacterial brown spot on snap beans, swarms with a characteristic dendritic pattern on semisolid (0.4%) agar plates. Filamentation of swarming cells of B728a was not observed. Mutations in either the gacS (formerly lemA) or gacA gene of B728a eliminate the ability of this P. syringae isolate to swarm without obvious effects on bacterial motility. Three field isolates showed a similar dependence on gacS for swarming. Since gacS and gacA mutants are known to be deficient in N-acyl-L-homoserine lactone (acyl HSL) production, a mutant was constructed by disruption of the ahlI gene of B728a. This mutant did not make any acyl-HSL detectable by the Agrobacterium traG::lacZ reporter system, yet was unaffected in its ability to swarm. Other phenotypes of gacS and gacA mutations were similarly unaffected in the ahlI mutant. PMID- 10383989 TI - Molecular cloning and characterization of a locus responsible for O acetylation of the O polysaccharide of Legionella pneumophila serogroup 1 lipopolysaccharide. AB - Complementation experiments, Tn5 mutagenesis, and DNA sequencing were used to identify a locus (lag-1) that participates in acetylation of Legionella pneumophila serogroup 1 lipopolysaccharide. Nuclear magnetic resonance analyses of lipopolysaccharides from mutant and complemented strains suggest that lag-1 is responsible for O acetylation of serogroup 1 O polysaccharide. PMID- 10383990 TI - Characterization of the Absolute Crystal Polarity across Twin Boundaries in Gallium Phosphide Using Convergent-Beam Electron Diffraction. AB - : The measurement of absolute crystal polarity is crucial to understanding the structural properties of many planar defects in compound semiconductors. Grain boundaries, including twin boundaries, in the sphalerite lattice are uniquely characterized by the crystallographic misorientation of individual grains and the direction of the crystal polarity in domains adjoining the grain boundary. To evaluate crystal polarity in gallium phosphide (GaP), asymmetrical interference contrast in convergent-beam electron-diffraction (CBED) patterns was used to ascertain the nature and direction of polar bonds. The direction of the asymmetry in the electron diffraction reflections was correlated with the crystal polarity of a sample with known polarity. The CBED technique was applied to determine the polar orientation of grains adjoining Sigma = 3 coherent and lateral twin boundaries in polycrystalline GaP. PMID- 10383991 TI - Improvement in Quantitative X-ray Microanalysis of Biological Cryosections. AB - : The accuracy of Na measurements in biological cryosections has been improved through replacement of the 8 um Be window of the Si(Li) detector with a Super Atmospheric Thin Window (SuperATW) window mounted on a germanium electron microscope (GEM) detector. Greater accuracy of Ca measurements was also attained when a GEM detector instead of an Si(Li) detector was used for analysis of total Ca concentrations in cardiac muscle. Although the advantage of improved sensitivity for Na and Ca is lost in spot mode analysis because of the overriding contribution of biological variability, this advantage becomes important in X-ray mapping. We compared the two systems by analyzing cryosections of identical biological material under comparable conditions. We found that Si contamination in thin biological cryosections is unpredictable; it may be widespread and differ in degree in various cell compartments of the same cell. Hence, a correction for Si contamination should be included in the calculations of elemental concentrations. We suggest here a procedure for measuring and subtracting Si contamination from elemental spectra. PMID- 10383992 TI - Topographic Imaging of Chromium-Coated Frozen-Hydrated Cell and Macromolecular Complexes by In-Lens Field Emission Scanning Electron Microscopy. AB - : An in-lens Schottky field emission scanning electron microscope (SEM) combined with a transmission electron microscope (TEM)-type cold-stage and a chromium (Cr) sputter-coating system was developed to rapidly prepare and cryo-image biological specimens to attain accurate nanometer-level structural information. High resolution topographic images at high primary magnification (>/=200,000x) were digitally recorded with very short dwell times and without beam damage. Plunge freezing in ethane, followed by fracturing, Cr coating, and in-lens cryo-high resolution scanning electron microscope (HRSEM) imaging directly revealed macromolecular features of yeast cells, platelets, and cell-free elastin analogues. The "vitreous" nature of bulk water in its solid state appeared featureless in cryo-HRSEM images, suggesting that if ice crystals were present they would be Abstract The intensive rearing of various fish species in aquaculture has revealed intimate relationships between fish and bacteria that eventually may affect establishment of a "normal" mucosal microflora or result in disease epizootics. Interactions between bacteria and mucosal surfaces play important roles both at the egg and larval stages of marine fish. Bacterial adhesion and colonization of the egg surface occur within hours after fertilization. The diverse flora which eventually develops on the egg appears to reflect the bacterial composition and load of the ambient water, but species-specific adhesion at the egg surface may also play a role in development of the egg epiflora. Proteolytic enzymes produced by members of the adherent epiflora may cause serious damage to the developing egg and may also affect further adhesion of the epiflora. Ingestion of bacteria at the yolk sac stage results in establishment of a primary intestinal microflora which seems to persist beyond first feeding. Establishment of a gut microflora is likely to undergo several stages, resulting in an "adult" microflora weeks to months after first feeding. Ingested bacteria may serve as an exogenous supply of nutrients or essential factors at an early life stage. Early exposure to high bacterial densities is probably important for immune tolerance, and thus for the establishment of a protective intestinal microflora. Successful rearing of early life stages of several marine fish species depends on knowledge of the complex interactions among the cultured organisms and the bacterial communities which develop at the mucosal surfaces and in the ambient water and rearing systems. The routine use of antibiotics during rearing of fish larvae is not advisable, since it may increase the risk of promoting antibiotic resistance and adversely affect the indigenous microflora of the larvae. The use of probiotics has proven advantageous in domestic animal production, and the search for effective probiotics may have a great potential in aquaculture of marine organisms. Bacteria with antagonistic effects against fish pathogens have been successfully administered to several fish species, resulting in decreased mortality or increased growth rate.http://link.springer-ny.com/link/service/journals/00248/bibs/38n1p1.html PMID- 10384007 TI - Ultraviolet Radiation (UVR) Sensitivity Analysis and UVR Survival Strategies of a Bacterial Community from the Phyllosphere of Field-Grown Peanut (Arachis hypogeae L.). AB - > Abstract The short-term population dynamics of the culturable bacterial community from field-grown peanut (Arachis hypogeae L.) was analyzed over three 2 day periods. As in other phyllosphere studies, significant numbers of pigmented organisms were detected, suggesting the importance of pigmentation in the colonization of this habitat. Isolates were grouped according to pigmentation (orange, pink, yellow, nonpigmented), and the sensitivity of each isolate in the collection (n = 617) to ultraviolet radiation (UVR) was determined as the minimal inhibitory dose (MIDC) of UVR that resulted in an inhibition of growth compared to an unirradiated control. The majority of isolates recovered (56.1%) had an MIDC equal to or exceeding that of Pseudomonas syringae 8B48, a known UV-tolerant strain. Among pigmentation groups, the mean MIDC of pink- and orange-pigmented isolates was significantly greater than that of yellow- or nonpigmented isolates at each sampling time of day. Identification of 213 of the isolates using fatty acid methyl ester analysis indicated that a large proportion of the isolates were gram-positive, with Bacillus spp. alone accounting for 35.7% of the total. The genus Curtobacterium contained the largest percentage of highly UVR-tolerant strains. Nonpigmented mutants of four Curtobacterium strains were selected following ethyl methane sulfonate mutagenesis; these nonpigmented mutants were significantly altered in survival following irradiation with UV-A wavelengths. The strategy of avoidance of UVR through colonization of the abaxial leaf surface was evaluated on three separate occasions by leaf imprint sampling. Only 3 of 120 leaves (2.5%) contained larger bacterial populations on the adaxial surface, indicating that colonization of the abaxial leaf surface is important to phyllosphere survival. Our results indicate that tolerance to UVR is a common phenotype among phyllosphere bacteria, suggesting that solar radiation has a strong influence on the microbial ecology of the phyllosphere.http://link.springer ny.com/link/service/journals/00248/bibs/38n1p27.html PMID- 10384008 TI - Influence of an Elevated Atmospheric CO2 Content on Soil and Rhizosphere Bacterial Communities Beneath Lolium perenne and Trifolium repens under Field Conditions. AB - > Abstract The increase in atmospheric CO2 content alters C3 plant photosynthetic rate, leading to changes in rhizodeposition and other root activities. This may influence the activity, the biomass, and the structure of soil and rhizosphere microbial communities and therefore the nutrient cycling rates and the plant growth. The present paper focuses on bacterial numbers and on community structure. The rhizospheres of two grassland plants, Lolium perenne (ryegrass) and Trifolium repens (white clover), were divided into three fractions: the bulk soil, the rhizospheric soil, and the rhizoplane-endorhizosphere. The elevated atmospheric CO2 content increased the most probable numbers of heterotrophic bacteria in the rhizosphere of L. perenne. However, this effect lasted only at the beginning of the vegetation period for T. repens. Community structure was assessed after isolation of DNA, PCR amplification, and construction of cloned 16S rDNA libraries. Amplified ribosomal DNA restriction analysis (ARDRA) and colony hybridization with an oligonucleotide probe designed to detect Pseudomonas spp. showed under elevated atmospheric CO2 content an increased dominance of pseudomonads in the rhizosphere of L. perenne and a decreased dominance in the rhizosphere of T. repens. This work provides evidence for a CO2-induced alteration in the structure of the rhizosphere bacterial populations, suggesting a possible alteration of the plant-growth-promoting-rhizobacterial (PGPR) effect.http://link.springer-ny.com/link/service/journals/00248/bibs/38n1p39.html PMID- 10384009 TI - Decomposition and CO2 Evolution from Standing Litter of the Emergent Macrophyte Erianthus giganteus. AB - > Abstract Decomposition of standing litter of the emergent macrophyte Erianthus giganteus (plumegrass) was quantified in a small freshwater wetland in Alabama, USA. Living green shoots of E. giganteus were tagged and periodically retrieved for determination of leaf and culm mass loss, litter-associated fungal biomass (ergosterol), and nitrogen and phosphorus concentrations. Laboratory studies were also conducted to examine the effects of plant litter moisture content and temperature on rates of CO2 evolution from plant litter. Culm and leaf material lost 25 and 32% AFDM, respectively, during plant senescence and early litter decay. Fungal biomass, as determined by ergosterol concentrations, increased significantly in both leaf and culm litter during decomposition, with maximum biomass accounting for 3.7 and 6.7% of the total detrital weight in culm and leaf litter, respectively. Spatial differences in fungal biomass were observed along the culm axis, with upper regions of the culm accumulating significantly greater amounts of fungal mass than basal regions (p < 0.01, ANOVA). Rates of CO2 evolution from both leaf and culm litter increased rapidly after wetting (0 to 76 ug CO2-C g-1 AFDM h-1 within 5 min). In addition, rates of CO2 evolution from water saturated culms increased exponentially as the temperature was increased from 10 to 30 degrees C. These results provide evidence that considerable microbial colonization and mineralization of standing emergent macrophyte litter can occur before collapse of senescent shoot material to the water and sediment surface.http://link.springer-ny.com/link/service/journals/00248/bibs/38n1p50.html PMID- 10384010 TI - Diversity of Microorganisms Isolated from Amber. AB - > Abstract Claims that organisms can be cultured from amber, if substantiated, would be significant contributions to our understanding of the evolution, tenacity, and potential spread of life. Three reports on the isolation of organisms from amber have been published. Cano and Borucki recently reported the isolation of Bacillus sphaericus and Lambert et al. have described a new species designated Staphylococcus succinus from 25-40 million year old Dominican amber. These characterized organisms were phylogenetically distant from extant relatives and the Staphylococcus sp. sufficiently far removed from other extant staphylococci to be considered a new species. Here we report the culture of bacteria from Dominican and previously untested 120 million year old Israeli (Lebanese lode) amber. Twenty-seven isolates from the amber matrix have been characterized by fatty-acid profiles (FAME) and/or 16S rRNA sequencing. We also performed a terminal restriction fragment pattern (TRF) analysis of the original amber before prolonged culture by consensus primer amplification of the 16S rRNA followed by restriction enzyme digestion of the amplicons. Sample TRFs were consistent with a sparse bacterial assemblage and included at least five of the isolated organisms. Finally, we microscopically mapped the internal topography of an amber slice.http://link.springer ny.com/link/service/journals/00248/bibs/38n1p58.html PMID- 10384011 TI - Stimulation of Diesel Fuel Biodegradation by Indigenous Nitrogen Fixing Bacterial Consortia. AB - > Abstract Successful stimulation of N2 fixation and petroleum hydrocarbon degradation in indigenous microbial consortia may decrease exogenous N requirements and reduce environmental impacts of bioremediation following petroleum pollution. This study explored the biodegradation of petroleum pollution by indigenous N2 fixing marine microbial consortia. Particulate organic carbon (POC) in the form of ground, sterile corn-slash (post-harvest leaves and stems) was added to diesel fuel amended coastal water samples to stimulate biodegradation of petroleum hydrocarbons by native microorganisms capable of supplying a portion of their own N. It was hypothesized that addition of POC to petroleum amended water samples from N-limited coastal waters would promote the growth of N2 fixing consortia and enhance biodegradation of petroleum. Manipulative experiments were conducted using samples from coastal waters (marinas and less polluted control site) to determine the effects of POC amendment on biodegradation of petroleum pollution by native microbial consortia. Structure and function of the microbial consortia were determined by measurement of N2 fixation (acetylene reduction), hydrocarbon biodegradation (14C hexadecane mineralization), bacterial biomass (AODC), number of hydrocarbon degrading bacteria (MPN), and bacterial productivity (3H-thymidine incorporation). Throughout this study there was a consistent enhancement of petroleum hydrocarbon degradation in response to the addition of POC. Stimulation of diesel fuel biodegradation following the addition of POC was likely attributable to increases in bacterial N2 fixation, diesel fuel bioavailability, bacterial biomass, and metabolic activity. Toxicity of the bulk phase water did not appear to be a factor affecting biodegradation of diesel fuel following POC addition. These results indicate that the addition of POC to diesel-fuel-polluted systems stimulated indigenous N2 fixing microbial consortia to degrade petroleum hydrocarbons.http://link.springer ny.com/link/service/journals/00248/bibs/38n1p69.html PMID- 10384012 TI - Microfungal Community Changes in Rodent Food Stores over Space and Time. AB - > Abstract This paper reports the changes over time in the microfungal communities that inhabit three rodent species' food stores at two climatically different locations. Results reveal that microfungal diversity values calculated from above-ground food stores are highest in the more commonly disturbed portions of the rodent dens. Interactions among food-inhabiting microbes and between the rodents and food-inhabiting microbes also appear to influence the microfungal communities within the rodent dens. For example, our data suggest that transport by animal vectors, and not by air currents, is more effective at dispersing microbial propagules. Furthermore, although fungal communities inhabiting food stores within dens varied in composition and diversity over time, standardized substrates (sorghum seeds) simultaneously placed within the food stores converged in microfungal composition the longer they were left within the dens. We hypothesize that animal vectors, including rodents, make neighboring fungal communities more alike by introducing similar communities of microbes, which in turn initiate a cascade of biological interactions that, over time, result in similar microfungal communities inhabiting newly stored food items.http://link.springer-ny.com/link/service/journals/00248/bibs/38n1p79.html PMID- 10384014 TI - Mammaplasty with a periareolar dermal cloak for glandular support. AB - Analyzing the main surgical element of mammaplasty, almost all procedures incorporate a smaller or bigger dermal flap. The periareolar dermal cloak is a dermis flap corresponding to the skin pattern and pedicled to the nipple areola. The shape of the flap can be tailored as required but 2 cm of the dermis flap around the nipple should not be touched. The periareolar dermal flap has been used as a cloak; this dermal cloak is suitable for positioning the nipple and covering a part of the glandular tissues with support. With fastening of the cloak, a better tone of the breast tissues can be achieved. Mastopexy, reduction mammaplasty presented by technical detail of dermal cloak positioning and glandular support, has been done in 178 breast operations since 1992. The dermal cloak technique was used in 114 cases. The technique, clinical results, advantages, disadvantages, and complications are discussed. PMID- 10384013 TI - The role of the computer imaging system in modern aesthetic plastic surgery. AB - The author describes his 10-year experience of using computer imagery for the preparation of surgical procedures and their predictability. The final results prove to be comparable with those programmed, in particular, in the case of rhinoplasty and liposuction. In the case of operations for rejuvenation, the computer is of some help. However, for the patient, the clinical demonstration in front of a mirror is more effective. As for surgical interventions on the breast, whether for enlargement or for reduction, it is very difficult to anticipate the final result with a computer. PMID- 10384015 TI - Psychosocial impact of cosmetic rhinoplasty. AB - The psychosocial impact of cosmetic rhinoplasty in Scandinavia is poorly investigated. Therefore a study was undertaken utilizing a mailed audit covering self-percepted experiences before, during, and after surgery. A total of 67 of 80 patients responded to the questionnaire (84%), on average 18 months after surgery. The mean age was 31 years (range, 16-63 years) and the M/F ratio was 20/44 among the 64 patients analyzed, half of whom were of foreign extraction. The self-report disclosed that a majority of the patients had been preoccupied with their noses since puberty and that the mental awareness of nasal stigmatization was given mainly by the mirror (58%), not by others. Despite the early exclusion of patients with possible body dysmorphic disorder, almost one fourth had a severe complex, and one-third felt socially inhibited by their noses and avoided being looked at from certain angles. Surgery was not a significant problem even though it was performed under local anesthesia and i.v. sedation. The great majority (91%) were satisfied with the result and 89% would recommend the procedure to others. More than 60% felt more self-confident and perceived life as being easier. To conclude, successful rhinoplasty may change preoccupied patients' lives, because the majority simply stopped thinking about their noses. PMID- 10384016 TI - A case of human adjuvant disease after augmentation rhinoplasty. AB - Human adjuvant disease (HAD) is an autoimmune syndrome which is caused by prolonged hypersensitization of injected foreign materials. Usually, this occurs after mammary augmentation with foreign materials. We report a rare case of HAD after rhinoplasty with silicone injection. Thirty years ago, the patient underwent augmentation rhinoplasty with silicone injection. We removed the silicone and grafted the area with fascia lata. After the operation, local and systemic symptoms improved. PMID- 10384017 TI - Cervical rhytidectomy. AB - Based on anatomical and clinical considerations, a new classification of the six most common neck problems is presented. In general there are three types of necks lean, fatty, and medium-which can involve three kinds of tissue-skin, muscle, and fat-that develop wrinkles, laxities, and adiposities. Different magnitudes and combinations of these problems are observed in these three kinds of patients. Medial plication of the platysma is emphasized as a natural way to deal with neck bands. PMID- 10384018 TI - A ten-year experience using polyurethane-covered breast implants. AB - The purpose of this study is to disclose the low incidence (0.98%) of capsular contracture using polyurethane-covered silicone gel breast implants. Four hundred seven surgical interventions were performed during the 10-year period, 404 for hypomastia and 3 for breast reconstruction. PMID- 10384019 TI - A review of the literature on the etiology of capsular contracture and a pilot study to determine the outcome of capsular contracture interventions. AB - The etiology of capsular contracture is unclear and probably multifactorial. This review covers the literature on several proposed contracture factors, including filler material, implant placement, surface texture, and bacterial infection. The pilot study's goal was to test the feasibility of a data collection form, which could be used in a scaled-up study analyzing multiple surgeon's records. The goal of the expanded version of this study will be to determine the efficacy of available interventions for capsular contracture, including surveillance. The Breast Implant Public Health Project, LLC (BIPHP), piloted a retrospective review of outcomes in women who had interventions to relieve capsular contracture or had chosen a wait-and-watch approach. An evaluation of the efficacy of various treatments can help women decide if they want to pursue treatment at all and, if so, which treatment might offer them the best solution. BIPHP researchers (E.E.A., M.E.) developed a data collection form after reviewing records of three surgeons (B.C., W.P., V.L.Y.). During the data collection using the same records, we tested a randomization process to identify women with capsular contracture who underwent various interventions, including a wait-and-watch strategy, and those who had no mention of any intervention or waiting approach. Data were gathered on a total of 90 breasts with capsular contracture (scored Baker I-IV or qualitatively), of which 45 underwent a total of 102 interventions for capsular contracture. Interventions were classified as "closed capsulotomy," "surgical," or "watchful waiting." Closed capsulotomy was performed most often (47%), followed by surgery (29%) and watchful waiting (21%). Presurgical Baker scores averaged higher in breasts that underwent surgery (3.1) than for watchful waiting (2.5) or closed capsulotomy (2.3). Though closed capsulotomies had 100% of outcomes scoring "improved" or "same," 58% of the breasts underwent the procedure more than once, suggesting that the favorable outcome was short-lived. The wait and-watch approach resulted in scores of either "same" or "worse"; surgery (open capsulotomy, repositioning, or capsulectomy) resulted in 79% improved, 16% same, and 5% worse outcomes in breasts with outcomes listed. In all intervention procedure categories, outcomes were frequently unavailable; they were noted only 60% of the time (52/87). The missing 40% may have resulted from the doctor's failure to note it in the chart, satisfied patients not returning for additional treatment, or dissatisfied patients seeking treatment elsewhere. Generally, the data collection forms and procedures were workable; however, we uncovered issues to address in the scale-up of this pilot study: (1) the outcome report rate was 60%; (2) though Baker scores are commonly used to evaluate the degree of capsular contracture, it seems that grade I may have different meanings for different surgeons, which would need to be clarified; (3) participating surgeons will need to divulge standard-of-care items that they may not have included in medical records, but routinely performed (e.g., patient massage, use of prophylactic antibiotics); and (4) records were initially separated by "implant," then researchers realized that a more useful collection would be by "breast." The latter approach captures the history of the breast in one record, which may be more important to contracture than the differences in implants. With the modifications discussed, the study can be scaled up to encompass as many records as necessary to achieve robust statistical power. These data will add to the existing literature regarding factors associated with capsular contracture and identify factors that affect the successful outcome of capsular contracture interventions. PMID- 10384020 TI - Factors associated with breast implant rupture: pilot of a retrospective analysis. AB - This pilot study's goal was to test the feasibility of a data collection form which will be used in a scale-up study analyzing multiple surgeons' records. The goal of this expanded study will be to develop identifying factors for women who are at greater risk for having ruptured implants and, if necessary, target them for screening, surveillance, or intervention. In the pilot study, we compared factors associated with implant rupture in women with and without rupture. Similar studies have considered one or a few factors at a time and, generally, have given little attention to implant generation. We developed a data collection form after reviewing records of three surgeons. A total of 92 records was collected and analyzed. An important feature in the pilot was to compare the results of patients whose implants the surgeons had both implanted and explanted (n = 34) with those of patients whose implants the surgeons had only explanted (n = 55) (unknown = 3). This comparison could show if including all explantation patients in a surgeon's practice would bias the sample; however, based on this pilot data, concerns regarding this type of bias seem to be minimal. Similar amounts of data (e.g., implant information, history of capsular contracture, etc.) were collectable on patients whose surgeons both implanted and explanted them (87%) and who had different surgeons for implantation and explantation (84%). Though the data from this limited sample cannot offer firm conclusions on rupture associations, a few factors stood out: size of implants (38. 3% of ruptured versus 15.9% of intact implants were 100-200 cm3), history of mammography (46.8% of ruptured versus 24.4% of intact had mammograms, which is likely due to older women with older implants having more mammograms), and history of closed capsulotomy (85.1% of ruptured versus 68.9% of intact). Interestingly, additional procedures performed on the breast (e.g., scar revision, wound repair, etc.) did not affect rupture: both the ruptured and the intact groups had an average of 1.7 procedures performed. The data collection form tested very well in this pilot study. Also, including all patients in the study sample, instead of excluding those who received their implants elsewhere, did not change the results. Though there are not enough data to draw any firm conclusions regarding rupture factors, the collection instrument was rigorously tested and should perform well in an expanded study. PMID- 10384021 TI - Vertical mammaplasty for breast reduction. AB - All of our breast reduction surgeries during the last 3 years have been done in such a manner that only the vertical scar is left. Due to placing a precise marking to obtain a very nice projected breast reduced to an appropriate size and with no complications, significant patient satisfaction has been achieved. The reduction of the scar is down to almost 40% of what it would have been with an inverted "T" scar. PMID- 10384022 TI - Lip augmentation with AlloDerm acellular allogenic dermal graft and fat autograft: A comparison with autologous fat injection alone. AB - Many surgical options exist for lip augmentation, none of which consistently provide safe, lasting, and predictable volume gains. We describe and evaluate the use of AlloDerm acellular allogenic dermal graft in combination with fat autograft and compare the postoperative results with those of autologous fat injection alone. Analysis of the preoperative and 1- and 3-month postoperative photographs was done using digital imaging software. Outcome measures included vermilion show and horizontal lip projection from the soft tissue pogonion subnasale plane. A 61% mean increase in vermilion show was observed in lips augmented with AlloDerm/fat injection, in comparison to a mean increase of 13% in lips augmented with fat injection alone. Lip projection demonstrated a mean increase of 1 mm in AlloDerm/fat lips at 3 months. Postoperatively, no evidence of resorption was seen in lips augmented with AlloDerm/fat between the 1- and the 3-month follow-ups, however, a 9% decrease in vermilion show occurred in lips augmented with fat injection over the same period. No complications occurred in either group. We conclude that AlloDerm in conjunction with autologous fat injection constitutes a safe, reliable, and lasting method of lip augmentation providing increased vermilion show compared to that with autologous fat injection alone. PMID- 10384023 TI - Malignant tumors associated with nevus sebaceous: therapeutic consequences. AB - Nevus sebaceous has been considered a relatively infrequent and unimportant congenital hamartoma for plastic surgeons unless the lesions are so big that they require a demanding defect closure. As the dignity of such tumors is primarily benign and the malformed sebaceous glands are localized abnormally high in the dermis, the temptation is appealing not to excise these tumors any more but to eradicate them by laser beam therapy. Yet a nevus sebaceous not only affects sebaceous glands but includes various other malformations of the affected skin and its appendages. In addition, different malignant tumors may occur in nevus sebaceous, even in children and young adults. We encountered 4 such malignant tumors of 18 nevi sebaceous operated on from 1989 to 1997. All nevi had been unsuspicious macroscopically. In three patients, one of them only 15 years old, an associated basal cell carcinoma was found. In the fourth patient there was a mixture of three additional tumors, a cystadenoma, a keratoacanthoma, and a basal cell carcinoma, besides the sebaceous malformations. These findings have two consequences: first, to continue surgical treatment of nevus sebaceous instead of dermabrasion or dermablation and to have the specimen examined histologically and, second, to excise such tumors as early in childhood as possible. PMID- 10384025 TI - FORUM: Environmental Testing, Official Methods, and Attitudes Toward Noncompliance. AB - / The US Environmental Protection Agency (EPA) has established longstanding regulations that prescribe the analytical procedures to be followed when parties submit information pursuant to regulatory programs. However, problems associated with unnecessary or irrational testing requirements along with the difficulty in obtaining approval of alternate procedures, has resulted in widespread noncompliance with those regulations. For the most part,agencies have tolerated this attitude of noncompliance and have in some ways contributed to it by adopting testing requirements that make little or no sense. Unfortunately, this complacency leaves agencies and regulated parties vulnerable to legal problems, such a court challenges to environmental permits. Regulatory agencies should confront the problems that have led to pervasive noncompliance and amend regulations to reflect that concerns about having useful data frequently override concerns about the national uniformity of test methods. In addition, the regulated community should be more mindful of the legal enforceability of promulgated testing requirements and affirmatively address those concerns with the promulgating agencies.KEY WORDS: Analytical chemistry; Biochemical oxygen demand; Compliance attitudes; Environmental lawhttp://link.springer ny.com/link/service/journals/00267/bibs/24n2p141.html PMID- 10384024 TI - A nondeforming rhytidectomy incision. AB - A new incision for facial rhytidectomy is presented that completely avoids deformation of the frame of the hair and allows the hair to be combed back without showing the scar, which becomes almost completely invisible in most patients if the incision is made at exactly the specified level. Several authors have tried to maintain the normal hairline, but some of their incisions deform the frame of the hair on the sides, higher than the level of the outer corner of the eye, and others go even higher than this point outside of the hairline, making the scar quite visible! The indications and contraindications for this operation and its designed incisions are discussed. These incisions encourage plastic surgeons not to remove any scalp in the rhytidectomy, but only the skin, because it is precisely the removal of scalp instead of skin that deforms the face, unless the amount of scalp tissue removed is very small. These same incisions are indicated for men. PMID- 10384026 TI - PROFILE: Integrated Environmental Management: The Foundations for Successful Practice. AB - / Integrated environmental management (IEM) is a holistic and goal-oriented approach to environmental management that addresses interconnections through a strategic approach. Although no models of IEM have emerged, practitioners throughout the world are forging ahead with the concept. The literature indicates that stakeholder collaboration and public involvement are central to operationalizingthis model, because this interaction produces a more integrated approach and generates support for implementation. However, it is not clear what steps and conditions are necessary for successful translation of IEM into operation. The author draws on twenty-three case studies from the United States and Australia, a survey of 285 Australian stakeholders and the literature to produce a framework for IEM. The framework identifies 20 elements that-if attained-will increase the likelihood of successful operationalization of IEM. These elements address structuring of an integrated approach, operation of stakeholder processes, and outputs and outcomes. The elements do not constitute a formula for success, but a generic set of attributes that constitute a foundation for effective practice.KEY WORDS: Integrated environmental management; Stakeholder collaboration; Ecosystem managementhttp://link.springer ny.com/link/service/journals/00267/bibs/24n2p151.html PMID- 10384027 TI - Cooperative Solutions for Sustainable Resource Management. AB - / Many environmental management issues can be defined as allocation problems, e.g., the allocation of rights to use common-pool resources or the allocation of the cost of regional resource development projects. The allocation methods developed in the area of cooperative n-person game theory are most appropriate for these problems because they focus on the conditions for engendering and sustaining the necessary cooperation among the involved stakeholders. These solution concepts seek to ensure that the allocation is based on some norm of equity and, most often, also to minimize the incentive for any player to defect from the cooperative venture. We illustrate these solution concepts with an application to a water resource project in Southern California. We argue how the rigorous mathematical nature of these solution concepts should not hinder their application to actual situations and how, with the use of heuristic rules and inexact notions of comparable worths, we can employ these concepts even in approximate fashion. We remind ourselves that the goal of such an endeavor is to convince stakeholders of the equity of a proposed solution and, in so doing, maximize the prospect of sustained cooperation. The alternative to cooperation, on the other hand, may be endless stalemate.KEY WORDS: Core; Game theory; Equity; Common-pool resources; Sustainabilityhttp://link.springer ny.com/link/service/journals/00267/bibs/24n2p167.html PMID- 10384028 TI - A Critical Review of Assumptions About the Prairie Dog as a Keystone Species. AB - / Prairie dogs (Cynomys spp.) have been labeled as keystone species because of their influence on biological diversity and ecosystem function. However, the validity of several assumptions used to support keystone status is questionable. We review the strength of the evidence and the magnitude of the prairie dog's effects on ecosystem structure and function. We use this review to reevaluate the keystone role for prairie dogs. Our goal is to encourage sound management of the prairie dog ecosystem by improving the ecological foundation of their keystone status. Our review confirms that prairie dogs affect a number of ecosystem-level functions but that their influence on prairie vertebrates may be less than previously suggested. Species richness and abundance patterns were variable among plants, mammals, and birds and were not consistently higher on prairie dog colonies compared to uncolonized areas. In addition, only nine of the 208 species listed in the literature as observed on or near prairie dogs colonies had quantitative evidence of dependence on prairie dogs. Abundance data indicated opportunistic use of colonies for an additional 20 species. A total of 117 species may have some relationship with prairie dogs, but we lacked sufficient data to evaluate the strength of this relationship. The remaining 62 species may be accidental or alien to the system. Despite our conclusion that some prairie dog functions may be smaller than previously assumed, collectively these functions are quite large compared to other herbivores in the system. We suggest that prairie dogs also provide some unique functions not duplicated by any other species and that continued decline of prairie dogs may lead to a substantial erosion of biological diversity and landscape heterogeneity across prairie and shrub-steppe landscapes. Thus, we concur that keystone status for prairie dogs is appropriate and may aid conservation efforts that help protect species dependent on prairie dogs and support other important ecosystem functions.KEY WORDS: Prairie dogs; Cynomys spp.; Keystone species; Ecosystem functions; Biological diversityhttp://link.springer ny.com/link/service/journals/00267/bibs/24n2p177.html PMID- 10384029 TI - RESEARCH: Evaluating Sustainability of Watershed Resources Management Through Wetland Functional Analysis. AB - / Unsustainable agricultural policies and water and soil resource schemes have drained two thirds of Mediterranean wetlands since 1920. An outstanding example is Karla in Greece, a former internationally important wetland that was drained in 1962 causing environmental, social, and water and soil problems. The objective of this study was to assess the functions and values of Karla, at three periods of its history, and to relate them to major events in the management of the water and soil resources of its watershed. Information on wetland and watershed features was collectedfrom historical records and field visits. The results showed that the wetland in its pristine state had performed five functions to a high degree, one (groundwater recharge) to a moderate degree, and one (flood storage) to a low degree. Flood-control works, uncontrolled pumping, etc., in 1936-1961 degraded all functions except microclimate modification while, the bird support function was moderately altered. Drainage works in 1962 left a very small artificially flooded wetland with only four functions performed to an insignificant degree. Value degradation followed function degradation. It was concluded that past resource management has been nonintegrated. No consideration was given to the multiple functions and values of Karla. Previous restoration proposals involved the reinstatement of one or two functions only. The appropriate restoration scheme for Karla must be multiobjective and based on the integrated resource management of its own and the neighboring watersheds.KEY WORDS: Soil and water resources; Sustainability; Watershed management; Wetland functional analysis; Karla wetlandhttp://link.springer ny.com/link/service/journals/00267/bibs/24n2p193.html PMID- 10384030 TI - Models to Assess the Risk of Snow and Wind Damage in Pine, Spruce, and Birch Forests in Sweden. AB - / Each year damage to forests caused by snow and wind causes high economic losses. In Sweden, approximately 4 million m3 are damaged annually by snow and wind, roughly corresponding to a value of US$150 million, and in Europe, the damage amounts to hundreds of millions of US dollars each year. To help to reduce these losses, tools for risk assessment within forest management have been developed. Predictions were developed of the risk of damage from snow and wind to Scots pine (Pinus sylvestris L.), Norway spruce [Picea abies (L.) Karst.] and Birch (Betula spp. L.) plots using tree, stand, and site characteristics. The data were obtained from 6756 permanent sample plots within the Swedish National Forest Inventory, which were inventoried twice at five-year intervals between 1983 and 1992. Input data for model development used measurements from the first inventory of tree characteristics for the largest sample tree, stand, and site data, and records of snow and wind damage from the second inventory. Models were developed for three different regions for pine- and spruce-dominated sites, while models for the whole country were developed for birch sites. In general the estimated proportion of damaged plots was highly overestimated (31.7%-56.2%), compared with the observed proportion of 3.4%-11.9%. The models for Norway spruce comprising tree, stand, and site data show the best predictability of damaged plots, with 60.6%-67.6% of plots correctly classified. It is concluded that the models developed can be used to detect sites with a high probability of damage from snow and wind, and thus be used as tools to reduce future damage and costs in practical forestry.KEY WORDS: Birch; Norway spruce; Risk assessment; Risk management; Scots pine; Silviculturehttp://link.springer ny.com/link/service/journals/00267/bibs/24n2p209.html PMID- 10384031 TI - Integrating Physical and Chemical Characteristics of Lakes into the Glacially Influenced Landscape of the Northern Cascade Mountains, Washington State, USA. AB - / A basic knowledge of the physical and chemical characteristics of lakes is needed by management to make informed decisions to protect water resources. In this study we investigated some of the physical and chemical characteristics of 58 lakes in alpine, subalpine, and forest vegetation zones in a natural area (North Cascades National Park Service Complex) between 1989 and 1993. The objectives of the study were to: (1) document the time of ice-out relative to lake elevation; (2) determine how a sharp climate gradient west and east of the hydrologic divide affected the time of ice-out for subalpine lakes; and (3) assess how lake water quality was associated with lake elevation, lake depth, and basin geology. As expected, lake ice-out times occurred earlier with decreasing elevation. East-slope subalpine lakes iced-out earlier than did west-slope subalpine lakes because the east slope of the study area was drier and warmer than the west slope. On average, the lakes were relatively cold, neutral in pH, and low in dissolved substances and concentrations of nitrogen and phosphorus. Although some shallow lakes (depth <10 m) exhibited the highest alkalinities, conductivities, and concentrations of phosphorus and nitrogen, most shallow lakes exhibited low values for these variables that were comparable to values observed in deep lakes. Geology did not play a major role in segregating the lakes based on water quality. Overall, lake temperature, pH, alkalinity, conductivity, and concentrations of total phosphorus and total Kjeldahl N increased with decreasing elevation. These changes in water quality with decreasing elevation in this temperate mountainous region corresponded with warmer air temperatures and increased vegetation biomass, soil depth and maturity, and dissolved substances and nutrients.KEY WORDS: Limnology; Mountain lakes; Water quality; North Cascades National Park Service Complex; National Park Servicehttp://link.springer ny.com/link/service/journals/00267/bibs/24n2p219.html PMID- 10384032 TI - Hydrological and Oceanographic Considerations for Integrated Coastal Zone Management in Southern Belize. AB - / The objectives of this study are to: (1) characterize the meteorology and hydrology of the Maya Mountain-Marine Area Transect in southern Belize, (2) employ a simple water balance model to examine the discharge rates of seven watersheds to Port Honduras, (3) test the validity of the hydrological model, (4) explore the implications of potential landscape and hydrological alterations, and (5) examine the value of protected areas. The southern coastal portion of the study area is classified as wet tropical forest and the remainder as moist tropical forest. Rainfall is 3000-4000 mm annually. Resulting annual freshwater discharge directly into Port Honduras is calculated at 2.5 x 10(9) m3, a volume equal to the basin. During the rainy season, June-September, 84% of the annual discharge occurs, which causes the bay to become brackish. Port Honduras serves as an important nursery ground for many species of commercially important fish and shellfish. The removal of forest cover in the uplands, as a result of agriculture, aquaculture, and village development, is likely to significantly accelerate erosion. Increased erosion would reduce soil fertility in the uplands and negatively affect mangrove, seagrass, and coral reef productivity in the receiving coastal embayment. Alternatively, the conservation of an existing protected areas corridor, linking the Maya Mountains to the Caribbean Sea, is likely to enhance regional sustainable economic development. This study aims to support environmental management at the scale of the "ecoscape"-a sensible ecological unit of linked watersheds and coastal and marine environments.KEY WORDS: Ecosystem management; Coastal zone management; Belize; Hydrologyhttp://link.springer ny.com/link/service/journals/00267/bibs/24n2p229.html PMID- 10384033 TI - Strategic Planning of Recycling Drop-Off Stations and Collection Network by Multiobjective Programming. AB - / Effective planning of solid-waste recycling programs is a substantial challenge to the current solid-waste management systems in Taiwan. Due to the rapid depletion of landfill space and the continuing delay in construction programs of municipal incinerators, solid-waste management strategies have to be reorganized in light of the success of recycling, recovery, and reuse of secondary materials. One of these efforts is how to effectively allocate recycling drop-off stations of appropriate size and how to design efficient collection-vehicle routing and scheduling programs in the solid waste collection network. This management strategy is particularly important in the privatized system with recycling containers and material recovery facilities (MRFs) owned by one agency. This research seeks multiobjective evaluation of the trade-off between the number and size of drop-off stations, the population covered in the service network, the average walking distance to drop-off stations by the population, and the distance traveled by collection vehicles. It also illustrates the use of the multiobjective nonlinear mixed integer programming model to achieve such goals that are solved by the genetic algorithms (GA) in a geographical information system (GIS) platform. The case study shows the application potential of such a methodology in the city of Kaohsiung in Taiwan.KEY WORDS: Optimization analysis; Recycling; Solid waste managementhttp://link.springer ny.com/link/service/journals/00267/bibs/24n2p247.html PMID- 10384034 TI - ENVIRONMENTAL AUDITING: Utilization of Parking Lots in Hattiesburg, Mississippi, USA, and Impacts on Local Streams. AB - / Utilization of parking lots around Hattiesburg, Mississippi, USA, was examined to suggest mechanisms for reducing runoff into local streams. Stream health was assessed by a census of fish communities. We found that most parking lots were used well below capacity, even in the peak shopping period before the Christmas holiday. Levels of utilization were primarily determined by shopping center age and parking availability relative to the size of businesses served. Streams receiving runoff had high undercut banks and heavy sediment loads. Their fish faunas were depauperate compared with a similar-sized stream in a nearby rural area. To moderate the impact of runoff, we recommend that lot size be carefully calibrated to actual parking demand. New businesses should be encouraged to establish in existing, unused parking lots rather than constructing new parking areas.KEY WORDS: Drainage; Fish communities; Mississippi; Parking utilization; Runoff; Suburbs; Urbanizationhttp://link.springer ny.com/link/service/journals/00267/bibs/24n2p265.html PMID- 10384035 TI - Density and Distribution of Citemene Fields in a Miombo Woodland Environment in Zambia. AB - / The Bemba people of northern Zambia practice citemene shifting cultivation. We utilized Landsat satellite images from 1984 and 1992 to map the distribution of citemene fields in those two years and to assess changes in the spatial sustainability of citemene cultivation in a miombo woodland study area. The citemene fields were concentrated within about 5-6 km of roads. Between 1984 and 1992, there was a decrease in the number of fields, and an increase in the average distance from the road. These changes may have been due to the introduction of semipermanent maize farming in the intervening years. The estimated cycle times for woodland utilization may have been long enough to maintain sustainability under traditional citemene cultivation practices. However, cycle times may be too short to maintain sustainability within the woodlands along roads.KEY WORDS: Shifting agriculture; GIS; Sustainability; Citemene; Miombo; Zambiahttp://link.springer ny.com/link/service/journals/00267/bibs/24n2p273.html PMID- 10384036 TI - Genetic and physical maps of the mouse rd3 locus; exclusion of the ortholog of USH2A. AB - The rd3 retinal degeneration gene was previously mapped 10 +/- 2.5 cM distal to Akp1 on mouse Chromosome (Chr) 1 (Chang et al., 1993), a region that may be homologous to the locus of the human USH2A gene, which carries mutations responsible for Usher IIa retinal degeneration/hearing loss syndrome. An intercross from an Rb(11, 13)4Bnr(rd3/rd3) x C57BL/6J mating was set up, 428 F2 meioses were analyzed, and the rd3 gene was placed between the markers D1MIT292/D1MIT209 and D1MIT510, a distance of 1.40 +/- 0.57 cM. These flanking markers and the mouse ortholog of USH2A (Mush2a) were mapped in the T31 mouse radiation hybrid (RH) panel, with the result that D1MIT292/D1MIT209 and D1MIT510 were 7.9 cR3000 apart ( approximately 800 kb), and Mush2a was > 30 cR3000 proximal to the pair, excluding it from the rd3 locus. A contig spanning the rd3 locus and consisting of 2 YACs and one BAC was generated, and Mush2a was absent from it, confirming its exclusion from the locus. Comparison of adjacent marker pairs in the Whitehead genetic map and our genetic map showed some discrepancies in order of markers and genetic distances. Comparison of our genetic map and the RH map showed some highly skewed relationships between genetic and physical distances. PMID- 10384037 TI - Genomic organization and expression analysis of the mouse qkI locus. AB - qkI, encoding a KH domain-containing RNA binding protein, has been isolated as a candidate gene for the mouse neurological mutation quaking. Here, we describe detailed studies on its genomic structure and expression pattern. We isolated approximately 1 Mb of genomic region containing the quaking locus and determined its genomic organization. The qkI locus contains at least 9 exons spanning approximately 65 kb of DNA. It gives rise to six distinct transcripts encoding, theoretically, five different protein isoforms. Exons 1 through 4 are shared by all the transcripts, whereas coding exons and two distinct 3'-UTRs downstream to the exon 4 are differentially utilized. One isoform has a truncated KH domain and may act as an antagonist to the others. These findings and identification of a single transcription initiation site suggest that differential expression of each transcript is regulated by alternative splicing. Expression of each alternative transcript and protein product was also examined. Two types of transcripts, 5 kb A and B, are most abundant in the brain of newborn mice and are gradually downregulated thereafter. In contrast, the other three messages, 6 kb, 7 kb-A and B, increase as myelination proceeds and peak at 2 weeks of age, corresponding to the most active stage of myelination. Although the qkI messages and their products are abundant in brain and heart, a lower level of expression was found in various other tissues tested. Alternative transcripts that share the same 3' UTR showed very similar expression patterns, suggesting a regulatory role of the 3'-UTRs in qkI gene expression. PMID- 10384038 TI - T-cell proliferative response is controlled by loci Tria4 and Tria5 on mouse chromosomes 7 and 9. AB - Lymphocytes of mouse strains BALB/cHeA (BALB/c) and STS/A (STS) differ in their response to CD3 antibody (anti-CD3). We analyzed the genetic basis of this strain difference, using the Recombinant Congenic Strains (RCS) of the BALB/c-c-STS/Dem (CcS/Dem) series. Each of the 20 CcS/Dem strains carries a different, random combination of 12.5% genes from the nonresponding strain STS and 87. 5% genes of the intermediate responder strain BALB/c. Differences in the magnitude of anti CD3-induced response among CcS/Dem strains indicated that in addition to Fcgamma receptor 2 (Fcgr2) other genes are involved in the control of this response as well, and we have already mapped loci Tria1 (T cell receptor-induced activation 1), Tria2, and Tria3. In order to map additional Tria genes, we tested F2 hybrids between the high responder RC strain CcS-9 and the low responder strain CcS-11. Proliferation in complete RPMI medium without anti-CD3 is controlled by locus Sprol1 (spontaneous proliferation 1) linked to the marker D4Mit23 on Chr 4. At concentration 0.375 microg/ml anti-CD3 mAb, the response was controlled by a locus Tria4, which maps to the marker D7Mit32 on Chr 7. The response to the higher concentration of mAb, 3 microg/ml, was controlled by Tria5, which mapped to the marker D9Mit15 on Chr 9. Anti-CD3 is being used for modulation of lymphocyte functions in transplantation reactions and in cancer treatment. Study of mechanisms of action of different Tria loci could lead to better understanding of genetic regulation of these reactions. PMID- 10384039 TI - A gene map of the rat derived from linkage analysis and related regions in the mouse and human genomes. AB - We report the localization by linkage analysis in the rat genome of 148 new markers derived from 128 distinct known gene sequences, ESTs, and anonymous sequences selected in GenBank database on the basis of the presence of a repeated element. The composite linkage map of the rat contributed by our group integrates mapping information on a total of 370 different known genes, ESTs, and anonymous mouse or human sequences, and provides a valuable tool for comparative genome analysis. 206 and 254 homologous loci were identified in the mouse and human genomes respectively. Our linkage map, which combines both anonymous markers and gene markers, should facilitate the advancement of genetic studies for a wide variety of rat models characterized for complete phenotypes. The comparative genome mapping should define genetic regions in human likely to be homologous to susceptibility loci identified in rat and provide useful information for the identification of new potential candidates for genetic disorders. PMID- 10384040 TI - Linkage mapping of rat chromosome 5 markers generated from chromosome-specific libraries. AB - Seventy-six novel microsatellite markers with various simple sequence repeat (SSR) motifs are reported in this paper. They were generated on the basis of non radioactive library screening procedures from flow-sorted rat Chromosome (Chr) 5 specific DNA, and were mapped in three rat backcross populations. Fifty-four of these markers mapped to Chr 5, while the other 22 mapped to other chromosomes of the rat genome. The marker D3Uwm8 is a new microsatellite marker for the rat syndecan 4 (ryudocan) gene. A genotyping protocol based on agarose gel electrophoresis is also provided in this paper. PMID- 10384041 TI - Deletion in the beige gene of the beige rat owing to recombination between LINE1s. AB - We have determined the molecular genetic basis of the rat beige mutant, a model for human Chediak-Higashi syndrome. Deletion of a 578-bp sequence, which led to a frame shift and a presumably non-functional truncated BEIGE protein, was identified in beige cDNA. The beige rat had a deletion of about 20 kb of genomic DNA, including three exons, which constitute the deleted 578-bp cDNA fragment. LINE1s (Long Interspersed Nucleolar Element 1) were identified at the site of the deletion. Consensus recognition sequences for DNA topoisomerase I were clustered at the putative deletion junction sites in LINE1s. We conclude that the deletion in the beige gene mediated by recombination between LINE1s is the causative mutation in the beige rat. The recombination might have been induced by DNA topoisomerase I and the extensive sequence homology between LINE1s. PMID- 10384042 TI - Genomic assignment of the warfarin resistance locus, Rw, in the rat. AB - The locus responsible for resistance to the anticoagulants warfarin and bromadiolone (locus symbol Rw) was integrated into the rat (Rattus norvegicus) microsatellite genome map. Seventh-generation offspring of a segregating strain of rats heterozygous resistant to both compounds were tested with a blood clotting-response (BCR) test. No recombination between resistance to warfarin and bromadiolone was observed, indicating a common genetic basis. No recombinants were found between Rw and D1Arb18 (Myl2) located at the MIT-microsatellite map position 95.90 (SHRSP x BN F2-cross) or 82.24 (FHH x ACI F2-cross). Resistance segregated in a ratio expected for single, dominant gene responses. An equal number of females and males were resistant, but females retained higher percentage blood coagulation activities (PCA) after anticoagulant administration. Partial synteny between rat, mouse, and human suggests that Myl2 may serve as anchor to map the Rw homologs in mouse and human. PMID- 10384043 TI - Analysis of major repetitive DNA sequences in the dog (Canis familiaris) genome. AB - A systematic screening and analysis of repeated DNA sequences from a dog genomic library composed of small DNA inserts enabled us to characterize abundant canine repetitive DNA families. Four main families were identified: i) a group of highly repeated tRNA-derived short interspersed repetitive DNA elements (tRNA-SINEs); ii) another type of SINE-like element that was mainly found inserted into long interspersed repetitive elements (LINEs); iii) LINEs of the L1 type; and iv) satellite or satellite-like DNA. Surprisingly, no SINEs derived from 7SL RNA were found in the dog genome. These data should help in the analysis of canine DNA sequences and in the design of canine genome mapping reagents. PMID- 10384044 TI - A 5x genome coverage bovine BAC library: production, characterization, and distribution. AB - A bovine large-insert DNA library has been constructed in a Bacterial Artificial Chromosome (BAC) vector. The source DNA was derived from lymphocytes of a Jersey male. High-molecular-weight DNA fragments were produced by treatment with EcoRI/EcoRI methylase and cloned into the EcoRI site of pBACe3.6. In total, 157,240 individual BACs have been picked into 384-well plates. Approximately 190 randomly chosen clones have been characterized by Pulsed Field Gel Electrophoresis (PFGE) and have an average insert size of 105 kb, suggesting library coverage representing 5-6 genome equivalents. The frequency of clones without inserts is 4%. The chromosomal location of 51 BACs was studied by FISH; 3 showed more than one signal, indicating a chimerism frequency of roughly 6%. Approximately 50% of the clones in the library contain Simple Repeat Sequences (microsatellites), and 4% of the clones contain centromeric repeats. Insert stability was assessed by restriction digestion of DNA prepared from 20 clones after serial culture for one and three nights. Only one clone showed any evidence of an altered restriction pattern. Clones from 360 x 384-well plates (138,240 colonies) were gridded onto high-density membranes, and PCR superpools were produced from the same set of clones. Both membranes and superpools are available from the RZPD, Berlin (http://www.rzpd.de). PCR 4-D superpools have been prepared from an additional 23,000 clones. The library has been screened for a total of 24 single-copy sequences; positive clones have been obtained in all cases. PMID- 10384045 TI - A missense mutation in the bovine MGF gene is associated with the roan phenotype in Belgian Blue and Shorthorn cattle. AB - The Roan locus is responsible for the coat coloration of Belgian Blue and Shorthorn cattle. The solid-colored and white animals are homozygotes, and the roan animals, with intermingled colored and white hairs, are heterozygous. The roan phenotype was mapped to cattle Chromosome (Chr) 5 with microsatellites, and a candidate gene was proposed (Charlier et al. Mamm Genome 7, 138, 1996). PCR primers to the exons of this candidate gene, the steel locus or mast cell growth factor (MGF) were designed. Solid-colored and white animals were sequenced. A missense mutation at 654 bp (amino acid 193, Ala --> Asp) was detected in these two groups. A PCR-RFLP was designed to this single base pair change, and 143 animals in total (Belgian Blue, Shorthorn, and various other breeds) were screened. In addition, the Canadian Beef Cattle Reference Herd (http://skyway. usask.ca/ approximately schmutz) was used to verify Mendelian inheritance of this marker with the phenotypic inheritance of roan. Our data suggest that this mutation in the bovine MGF gene is responsible for the roan phenotype. PMID- 10384047 TI - Comparison of the human with the sheep genomes by use of human chromosome specific painting probes. AB - Human chromosome specific painting probes were hybridized on sheep (Ovis aries, 2n = 54) chromosomes by FISH. The painting results on sequentially stained RBA banded preparations demonstrated high degree of conserved regions between human and sheep genomes. A total of 48 human chromosome segments were detected in sheep chromosomes. Comparisons with sheep gene mapping data available and previous Zoo FISH data obtained in sheep, cattle, and river buffalo were performed. PMID- 10384046 TI - Reciprocal chromosome painting shows that the great difference in diploid number between human and African green monkey is mostly due to non-Robertsonian fissions. AB - We used reciprocal chromosome painting with both African green monkey (C. aethiops) and human chromosome specific DNA probes to delineate homologous regions in the two species. Probes were derived by fluorescence-activated chromosome flow sorting and then were reciprocally hybridized to metaphase spreads of each species. Segments in the size range of a single chromosome band were identified, demonstrating the sensitivity of the approach when comparing species that diverged more than 20 million years ago. Outgroup analysis shows that the great difference in diploid numbers between the African green monkey (2n = 60) and humans (2n = 46) is mainly owing to fissions, and the direction of change is towards increasing diploid numbers. However, most break points apparently lie outside of the centromere regions, suggesting that the changes were not solely Robertsonian as has been previously assumed. No reciprocal translocations have occurred in the phylogenetic lines leading to humans or African green monkeys. The primate paints established here are a valuable tool to establish interspecies homology, to define rearrangements, and to determine the mechanisms of chromosomal evolution in primate species. PMID- 10384048 TI - Characterization of several LINE-1 elements in Microtus kirgisorum. AB - Several LINE-1s have been isolated and characterized from genomic DNA of the vole, Microtus kirgisorum. Blot hybridization revealed specific restriction patterns of L1 elements in vole genomes. Rehybridization of the genomic blot with a cloned 5'-end fragment revealed two major bands indicating the presence of two different L1 subfamilies. The copy numbers are estimated for different parts of M. kirgisorum L1 elements. Data also demonstrate that most vole L1 elements are truncated at the 5'-end; however, in contrast to mouse, the ORF1 copy number is higher in vole. A difference between the substitution rates of the ORF1 5'-region (approximately 330 nucleotides) and the rest of the L1 coding regions is revealed. PMID- 10384049 TI - Mus and Peromyscus chromosome homology established by FISH with three mouse paint probes. AB - Fluorescence-labeled DNA probes constructed from three whole house mouse (Mus domesticus) chromosomes were hybridized to metaphase spreads from deer mouse (Peromyscus maniculatus) to identify homologies between the species. Mus Chr 7 probe hybridized strongly to the ad-centromeric two-thirds of Peromyscus Chr 1q. Most of Mus 3 probe hybridized principally to two disjunct segments of Peromyscus Chr 3. Mus Chr 9 probe hybridized entirely to the whole Peromyscus Chr 7. Three Peromyscus linkage groups were assigned to chromosomes, based on linkage homology with Mus. The data also are useful in interpretation of chromosomal evolutionary history in myomorphic rodents. PMID- 10384050 TI - A targeted screen to detect recessive mutations that have quantitative effects. AB - A key problem in mutagenesis research is developing methods that are sufficiently sensitive to detect a wide range of abnormal phenotypes. Major variants may be easy to identify, but it can be difficult to detect mutations that have subtle effects, particularly on a complex genetic background. This paper describes a targeted mutagenesis protocol with enough sensitivity to detect recessive mutations that have modest quantitative effects. The procedure relies on consomic inbred strains of mice-strains in which one homologous pair of chromosomes of an inbred strain has been replaced with the corresponding pair from a donor strain. Mice that carry the desired donor chromosome-the target of the screen-are mutagenized and bred back to the original recipient strain. The first-generation progeny (G1) that are heterozygous only for the donor chromosome are also bred back to the recipient strain. G2 animals that inherit nonrecombinant donor chromosomes are identified by genotyping. These animals may be backcrossed repeatedly to the recipient strain to dilute off-target mutations, but ultimately, nonrecombinant G2 animals are bred to each other. Their G3 progeny are genotyped at markers spaced at 5- to 10-cM intervals to identify mating pairs that are homozygous for shared segments of the mutagenized donor chromosome. Entire litters of G4 progeny that are homozygous for defined intervals are screened. By comparing phenotypes within and among litters of nearly isogenic G4 animals, mutations can be verified and simultaneously mapped with a precision of 5-10 cM. This method has the potential to consistently detect mutations that have effects on trait means of well under one standard deviation. PMID- 10384051 TI - Molecular characterization of Zfp54, a zinc-finger-containing gene that is deleted in the embryonic lethal mutation tw18. AB - Mapping studies have indicated that hundreds of Kruppel-type zinc-finger (ZNF) containing genes are present within the genomes of mammals. One-third or more of these genes contain a second, highly conserved element termed the Kruppel associated box (KRAB). In a recent study, several partial cDNA clones encoding distinct KRAB elements were mapped in mouse. One of these sequences, clone KRAB10, was assigned to mouse Chromosome (Chr) 9. We have isolated the complete coding portion of the KRAB10-containing gene and demonstrate that it is identical to Zfp54, one of several related ZNF genes that are located on mouse Chr 17 in the interval that is deleted in the tw18 mutation. Sequence analysis has shown that the KRAB A domain encoded by Zfp54 is highly similar to the comparable motif encoded by its nearest known neighbor, Zfp51. However, other portions of the related proteins, including their ZNF domains, are noticeably different in structure. Studies of Zfp54 expression revealed the presence of transcript in several adult tissues and in embryos throughout development; however, elevated levels of transcript were detected only in adult testis and in 7-day-old embryos. These data suggest that Zfp54 gene expression is under spatial and temporal control and may, therefore, play important roles in male germ cell and embryonic development. PMID- 10384053 TI - The spontaneous coat color mutant white nose (wn) maps to murine chromosome 15. PMID- 10384052 TI - Comparative analyses of the Dominant megacolon-SOX10 genomic interval in mouse and human. PMID- 10384054 TI - The canine copper toxicosis locus is not syntenic with ATP7B or ATX1 and maps to a region showing homology to human 2p21. PMID- 10384055 TI - Genomic organization of the human caspase-9 gene on Chromosome 1p36. 1-p36.3. PMID- 10384056 TI - cDNA cloning and the 5'genomic organization of Naip2, a candidate gene for murine Legionella resistance. PMID- 10384058 TI - The genomic organization of the histone clusters on human 6p21.3. PMID- 10384057 TI - Human-mouse comparative mapping of the genomic region containing CDK6: localization of an evolutionary breakpoint. PMID- 10384059 TI - Lung macrophage differentiation antigens in developing fetal and newborn rat lungs: A quantitative flow cytometric analysis with immunohistochemistry. AB - Relative deficiencies in the number and function of alveolar macrophages (AMs) are present immediately after birth and contribute to increased susceptibility to infection. We used immunohistochemical localization with a panel of monoclonal antibodies to macrophage differentiation markers to characterize lung macrophage antigens relevant to development, and we present here the first study to quantitate these markers using flow cytometric analysis. Rat lung macrophages undergo immunophenotypic maturation seen by a changing number (OX-1, ED-1, ED-2, and ED-9) and distribution (ED-2) of certain lung macrophage differentiation antigens. Quantification of these antibodies revealed that labeling for ED-1 and ED-9 was less on lavageable AMs from early postnatal days than on mature adult AMs. In vitro treatment of adult rat AMs with murine granulocyte-macrophage colony-stimulating factor produces a proliferating population of AMs with a high proliferation index and an immature phenotype, similar to that of newborn AMs. This in vitro model was useful in validating our quantitation of neonatal lung macrophage differentiation antigens. Our quantitative studies of potential markers of AM differentiation in the developing lung may help to focus the study of macrophages in the developing lung on specific cellular and molecular pathways that may control immunologic events during this critical period. PMID- 10384061 TI - Expression of C-C chemokines in bronchoalveolar lavage cells from patients with granulomatous lung diseases. AB - To determine the role of C-C chemokines in the pathogenesis of granulomatous lung diseases, we studied the mRNA levels of C-C chemokines, regulated on activation normal T expressed and secreted (RANTES), macrophage inflammatory protein (MIP) 1alpha, MIP-1beta, and monocyte chemoattractant protein (MCP)-1 in bronchoalveolar lavage (BAL) cells obtained from patients with sarcoidosis (n = 17), hypersensitivity pneumonitis (HP) (n = 4), and cryptogenic fibrosing alveolitis (CFA) (n = 10) using the reverse transcription-polymerase chain reaction (RT-PCR) technique. The mRNA levels of RANTES, MIP-1alpha, and MIP-1beta in BAL cells were significantly correlated with the lavaged lymphocyte proportion, and a significant inverse correlation was observed between the mRNA level of MIP-1beta and the CD4/CD8 ratio of lavaged lymphocytes. Among the three diseases, the mRNA levels of RANTES and MIP-1alpha were significantly higher in the patients with sarcoidosis or HP compared with those in the patients with CFA. The level of MIP-1beta mRNA was significantly higher in the HP patients compared with that in the patients with sarcoidosis or CFA. No significant differences were observed in the level of MCP-1 mRNA among the three diseases. Thus, RANTES and MIP-1alpha were suggested to be important in the pathogenesis of granulomatous inflammation in sarcoidosis and HP. MIP-1beta might play an important role in the pathogenesis of HP, mediating the recruitment of lymphocytes specific to HP. PMID- 10384060 TI - Budesonide down-regulates eosinophil locomotion but has no effects on ECP release or on H2O2 production. AB - Treatment of allergic asthma with inhaled corticosteroids results in local down regulation of proinflammatory cytokine synthesis and in marked decrease in tissue eosinophilia. Blood concentrations of inhaled corticosteroids, although significantly lower than those measured in the lung, may still have antiinflammatory effects on circulating eosinophils, reducing their ability to migrate. The aim of our study was to evaluate in vitro the activity of budesonide on blood eosinophils by measuring their chemotactic response, eosinophil cationic protein (ECP) release, and hydrogen peroxide (H2O2) production in the presence of different drug concentrations similar to those obtained at airway level (10(-8) and 10(-7) M) and at blood level (10(-10) and 10(-9) M). Partially purified blood eosinophils, isolated from 23 asthmatic subjects, were used to evaluate the activity of budesonide on: (1) chemotaxis toward the activated fifth component of complement (C5a, 0.1 microg/ml) or recombinant human (rh) interleukin (IL)-5 (200 pg/ml), (2) ECP release by cells stimulated with tetradecanoylphorbol acetate (TPA) and (3) H2O2 production by TPA-activated cells. The chemotactic response to C5a was down-regulated significantly by budesonide only by the highest concentrations tested (10(-8) and 10(-7) M); differently, budesonide was effective in inhibiting eosinophil migration toward rhIL-5, at all concentrations tested (p < 0.01, each comparison). By contrast, no drug-induced modifications were observed in ECP release or in H2O2 production (p > 0.05, each comparison). We conclude that concentrations of budesonide similar to those obtained in vivo are effective in inhibiting eosinophil locomotion but not in down-regulating the release of reactive oxygen species and granule-associated proteins. PMID- 10384062 TI - Bombesin inhibits apoptosis in developing fetal rat lung. AB - We have shown recently that apoptosis occurs during fetal and postnatal lung development. Our hypothesis that branching morphogenesis occurs through a delicate balance of cell proliferation and apoptosis predicts that substances that enhance branching of the airways would affect both cell proliferation and apoptosis in the lung. Bombesin-like peptides have a mitogenic effect on bronchial epithelium and fibroblasts, and bombesin has been shown to enhance branching morphogenesis in fetal lung. We used organ cultures of 16-day gestation fetal rat lung to study the effects of bombesin on apoptosis. Cultures were incubated in serumless medium alone or exposed to 1microM bombesin for 0-48 h. Levels of apoptosis were quantified using the TUNEL assay and expressed as percentage of apoptotic cells in paraffin sections of explants. Bombesin significantly inhibited apoptosis in fetal lung mesenchyme 48 h in culture by more than 50% (p < 0.05). The effects of bombesin on apoptosis were prevented completely if explants were exposed to the specific bombesin receptor antagonist, [D-Phe12]bombesin. To examine if the absence of serum in the media could have accounted for some of these effects, explants were cultured for 48 h in serumless medium, medium containing 10% fetal bovine serum, serumless medium with 1 microM bombesin, or medium containing both 10% fetal bovine serum and 1 microM bombesin. The addition of fetal bovine serum to the media reduced apoptosis significantly. The effect of fetal bovine serum on apoptosis was additive with bombesin. We conclude that bombesin inhibits apoptosis in developing fetal rat lung mesenchyme through its interaction with the bombesin receptor. PMID- 10384063 TI - Thrombin in the airways of asthmatic patients. AB - The mechanism of airway remodeling in asthmatic patients is poorly understood. Thrombin is a multifunctional protease that, in addition to its critical role in thrombotic processes, has also been described as inducing cellular and molecular events relevant to tissue remodeling. The present investigation was undertaken to evaluate the activity of thrombin in the sputum of asthmatic patients and its potential role in airway remodeling. The study population comprised 8 healthy subjects and 14 stable patients with bronchial asthma. The concentrations of thrombin, thrombin-antithrombin complex (TAT), and tissue factor were measured in the sputum of all subjects. The concentrations of thrombin (p = 0. 007), TAT (p = 0.01), and tissue factor (p = 0.02) in sputum were significantly higher in asthmatic patients than in healthy controls. The proliferative effects that sputum from asthmatic patients (p = 0. 01) and thrombin (p = 0.03) have on cultured human smooth muscle cells was inhibited significantly in the presence of recombinant hirudin, a specific thrombin inhibitor. Significant statistical correlation was observed between the degree of bronchial responsiveness and the sputum concentrations of thrombin (r = -0.8; p = 0.02) and TAT (r = -0.9; p = 0.01). The results of this study showed that increased thrombin generation occurs in the airway of patients with asthma and that it may play a role in the pathogenesis of airway remodeling. Further studies should be carried out to assess whether these findings are also observed in other airway diseases. PMID- 10384064 TI - Effect of terbutaline on exercise capacity and pulmonary function in patients with chronic obstructive pulmonary disease. AB - The objective of this study was to investigate the effects of a single dose of a beta2-agonist, terbutaline (Bricanyl Turbuhaler), on resting lung function and exercise capacity in patients with chronic obstructive lung disease. Using a double-blind, placebo-controlled, randomized crossover study and outpatients from a department of pulmonary medicine at a major inner-city hospital, we examined 26 individuals with chronic obstructive lung disease who met the criteria of 40% 70% of the individuals studied, and all individuals expressed at least one of the over-represented TCRAV families. Over represented conserved AV4A or AV7 sequences were also present in CD8+ T cells from most donors. The extent of TCRA sequence conservation is unparalleled. TCRAV4A, AV19, and AV24 sequences were invariant, although AV4A and AV19 transcripts contained N region additions. TCRAV24 transcripts derived from the direct juxtaposition of V and J gene segments. TCRAV7 sequences showed some diversity in two amino acids encoded at junctions of V and J gene segments. Although derivation of DN T cells with conserved TCRA chains is puzzling, the wide-spread expression of these unusual cells suggests an important function. PMID- 10384130 TI - Surfactant protein D binds to Mycobacterium tuberculosis bacilli and lipoarabinomannan via carbohydrate-lectin interactions resulting in reduced phagocytosis of the bacteria by macrophages. AB - Surfactant protein-D (SP-D) is a collectin produced in the distal lung airspaces that is believed to play an important role in innate pulmonary immunity. Naive immunologic responses to Mycobacterium tuberculosis (M.tb) are especially important in the lung, since entry of this inhaled pathogen into the alveolar macrophage is a pivotal event in disease pathogenesis. Here we investigated SP-D binding to M.tb and the effect of this binding on the adherence of M. tb to human macrophages. These studies demonstrate specific binding of SP-D to M.tb that is saturable, calcium dependent, and carbohydrate inhibitable. In addition to purified SP-D, SP-D in bronchoalveolar lavage fluids from healthy donors and patients with alveolar proteinosis also binds to M.tb. Incubation of M.tb with SP D results in agglutination of the bacteria. In contrast to its binding to M.tb, SP-D binds minimally to the avirulent Mycobacterium smegmatis. SP-D binds predominantly to lipoarabinomannan from the virulent Erdman strain of M.tb, but not the lipoarabinomannan from M. smegmatis. The binding of SP-D to Erdman lipoarabinomannan is mediated by the terminal mannosyl oligosaccharides of this lipoglycan. Incubation of M.tb with subagglutinating concentrations of SP-D leads to reduced adherence of the bacteria to macrophages (62.7% of control adherence +/- 3.3% SEM, n = 8), whereas incubation of bacteria with surfactant protein A leads to significantly increased adherence to monocyte-derived macrophages. These data provide evidence for specific binding of SP-D to M. tuberculosis and indicate that SP-D and surfactant protein A serve different roles in the innate host response to this pathogen in the lung. PMID- 10384131 TI - Genetic immunization of mice against Listeria monocytogenes using plasmid DNA encoding listeriolysin O. AB - The development of protective immunity against many intracellular bacterial pathogens commonly requires sublethal infection with viable forms of the bacteria. Such infection results in the in vivo activation of specific cell mediated immune responses, and both CD4+ and CD8+ T lymphocytes may function in the induction of this protective immunity. In rodent models of experimental infection with Listeria monocytogenes, the expression of protective immunity can be mediated solely by the immune CD8+ T cell subset. One major target Ag of Listeria-immune CD8+ T cells is the secreted bacterial hemolysin, listeriolysin O (LLO). In an attempt to generate a subunit vaccine in this experimental disease model, eukaryotic plasmid DNA expression vectors containing genes encoding either the wild-type or modified forms of recombinant LLO were generated and used for genetic vaccination of naive mice. Results of these studies indicate that the intramuscular immunization of mice with specifically designed plasmid DNA constructs encoding recombinant forms of LLO stimulates peptide-specific CD8+ immune T cells that exhibit in vitro cytotoxic activity. More importantly, such immunization can provide protective immunity against a subsequent challenge with viable L. monocytogenes, demonstrating that this experimental approach may have direct application in prevention of acute disease caused by intracellular bacterial pathogens. PMID- 10384132 TI - Protective immunity from naive CD8+ T cells activated in vitro with MHC class I binding immunogenic peptides and IL-2 in the absence of specialized APCs. AB - Ag-specific CTL can protect against tumors and some viral infections and may be useful for adoptive immunotherapy. Here, we show that purified CD8+ T cells from naive C57BL/6 mice can be primed in vitro with different immunogenic peptides, which bind to MHC class I gene products, and IL-2 to exhibit specific and MHC restricted effector function in vitro and in vivo protection against lymphocytic choriomeningitis virus infection and B16.F10 melanoma lung metastases. Limiting dilution assays in the absence of feeder cells with highly purified CD8+ T cells from two transgenic mice strains, each expressing a different MHC class I restricted TCR, indicated that only peptide and IL-2, but not TCR- cells, were required for the growth of naive CD8+ T cells. These alternative minimal requirements for the activation and expansion of specific CD8+ T lymphocytes, without the need for professional APC, may be exploited for adoptive immunotherapy. PMID- 10384133 TI - Schistosome-infected IL-4 receptor knockout (KO) mice, in contrast to IL-4 KO mice, fail to develop granulomatous pathology while maintaining the same lymphokine expression profile. AB - Th2 lymphocytes have been postulated to play a major role in the immunopathology induced by Schistosoma mansoni infection. Nevertheless, infected IL-4 knockout (KO) and wild-type (wt) mice develop egg granulomas comparable in size. To further investigate the function of the Th2 response in egg pathology we studied IL-4Ralpha-deficient mice, which are nonresponsive to both IL-4 and IL-13. In striking contrast to IL-4 KO animals, infected IL-4Ralpha KO mice developed only minimal hepatic granulomas and fibrosis despite the presence of CD3+ T cells in the residual egg lesions. Moreover, liver lymphokine mRNA levels in these animals and IL-4 KO mice were equivalent. In addition, infected IL-4Ralpha-deficient, IL 4-deficient, and wt animals developed similar egg Ag-specific IgG Ab titers, arguing that CD4-dependent Th activity is intact in KO mice. As expected, IFN gamma secretion was strongly up-regulated in mesenteric lymph node cultures from both groups of deficient animals, a change reflected in increased serum IgG2a and IgG2b Ab levels. Surprisingly, Th2 cytokine production in infected IL-4Ralpha KO mice was not abolished but was only reduced and resembled that previously documented in IL-4 KO animals. This residual Th2 response is likely to explain the ability of IL-4 KO mice to generate egg granulomas, which cannot be formed in IL-4Ralpha-deficient animals because of their lack of responsiveness to the same cytokine ligands. Taken together, these findings argue that tissue pathology in schistosomiasis requires, in addition to egg-specific CD4+ lymphocytes, a previously unrecognized IL-4Ralpha+ non-T cell effector population. PMID- 10384134 TI - Characterization of eosinophil adhesion to TNF-alpha-activated endothelium under flow conditions: alpha 4 integrins mediate initial attachment, and E-selectin mediates rolling. AB - The multistep model of leukocyte adhesion reveals that selectins mediate rolling interactions and that integrins mediate firm adhesion processes. In this study, the interaction between eosinophils and TNF-alpha-activated HUVEC (second or third passage) was studied under flow conditions (0.8 and 3.2 dynes/cm2). Especially the role of alpha 4 integrins on eosinophils and E-selectin on HUVEC was studied. Inhibition of the integrin alpha 4 chain on eosinophils reduced the number of firmly adhered resting eosinophils to TNF-alpha-stimulated endothelium by 43% whereas the percentage rolling cells increased 2.2-fold compared with untreated control eosinophils. Blocking of E-selectin on the endothelium reduced the number of adherent eosinophils by only 23% and 16%. In this situation, however, hardly any rolling adhesion was observed, and the few rolling cells showed a low rolling velocity. Blocking both alpha 4 integrin on eosinophils and E-selectin on HUVEC reduced the number of adhered eosinophils by 95%. P-selectin did not significantly participate in eosinophil adhesion to TNF-alpha-activated HUVEC. Inhibition of both alpha 4 integrins and beta 2 integrins on eosinophils resulted in a reduction of adhered cells by 65% and a 3-fold increase in percentage rolling cells. Taken together, these results clearly show that resting eosinophils preferentially use constitutively active alpha 4 integrins (alpha 4 beta 1, alpha 4 beta 7) for the first attachment to TNF-alpha-activated HUVEC. In addition, alpha 4 integrins and E-selectin work synergistically in eosinophil adherence to TNF-alpha-activated HUVEC. Although E-selectin is important for eosinophil rolling under these conditions, P-selectin plays only a minor role. PMID- 10384135 TI - Involvement of thioredoxin in rheumatoid arthritis: its costimulatory roles in the TNF-alpha-induced production of IL-6 and IL-8 from cultured synovial fibroblasts. AB - Thioredoxin (TRX) is a cellular reducing catalyst induced by oxidative stress and is involved in the redox regulation of transcription factors such as NF-kappaB. We found that the serum TRX concentration was elevated in patients with rheumatoid arthritis (RA) as compared with values from healthy individuals and patients with osteoarthritis (33.6 +/- 35.1 vs 11.8 +/- 6.6 ng/ml, p < 0.01). Moreover, the TRX concentration in the synovial fluid (SF) was much more elevated in RA patients than in osteoarthritis patients (103.4 +/- 53.3 vs 24.6 +/- 17.4 ng/ml, p < 0.001). Multiple regression analysis revealed that the serum C reactive protein value was better correlated with the linear combination of SF TNF-alpha and SF TRX values than with SF TNF-alpha alone, suggesting that TRX might play a subsidiary role in the rheumatoid inflammation. We thus examined the effect of TRX on the TNF-alpha-induced IL-6 and IL-8 production using rheumatoid synovial fibroblast cultures. The extents of IL-6 and IL-8 production in response to TNF-alpha were greatly augmented by TRX as compared with TNF-alpha alone. TRX alone did not have such effects. We also found that TRX appeared to accelerate the nuclear translocation of NF-kappaB, a major transcriptional regulator for production of IL-6 and IL-8 on stimulation with TNF-alpha. Consistent with these findings, the IkappaBalpha phosphorylation at Ser32 and its subsequent degradation in response to TNF-alpha was facilitated by TRX. These findings indicate that the elevated TRX concentration in SF of RA patients might be involved in the aggravation of rheumatoid inflammation by augmenting the NF kappaB activation pathway. PMID- 10384136 TI - Deficiency of 5-lipoxygenase abolishes sex-related survival differences in MRL lpr/lpr mice. AB - Leukotrienes, the 5-lipoxygenase (5LO) products of arachidonic acid metabolism, have many proinflammatory actions that have been implicated in the pathogenesis of a variety of inflammatory diseases. To investigate the role of LTs in autoimmune disease, we generated an MRL-lpr/lpr mouse line with a targeted disruption of the 5lo gene. MRL-lpr/lpr mice spontaneously develop autoimmune disease that has many features resembling human systemic lupus erythematosus, including sex-related survival differences; female MRL-lpr/lpr mice experience significant early mortality compared with males. Unexpectedly, we found that mortality was accelerated in male 5LO-deficient MRL-lpr/lpr mice compared with male wild-type MRL-lpr/lpr animals. In contrast, the 5lo mutation had no effect on survival in females. Mortality was also accelerated in male MRL-lpr/lpr mice that were treated chronically with a pharmacological inhibitor of LT synthesis. Furthermore, LT-dependent inflammatory responses are enhanced in male MRL-lpr/lpr mice compared with females, and the 5lo mutation has greater impact on these responses in males. Because immune complex-mediated glomerulonephritis is the major cause of death in MRL-lpr/lpr mice and has been related to arachidonic acid metabolites, we also assessed kidney function and histopathology. In male MRL lpr/lpr mice, renal plasma flow was significantly reduced in the 5lo-/- compared with the 5lo+/+ group, although there were no differences in the severity of renal histopathology, lymphoid hyperplasia, or arthritis between the groups. These findings suggest that the presence of a functional 5lo gene confers a survival advantage on male MRL-lpr/lpr mice and that, when 5LO function is inhibited, either genetically or pharmacologically, this advantage is abolished. PMID- 10384138 TI - Estrogen protects against cellular infiltration by reducing the expressions of E selectin and IL-6 in endotoxin-induced uveitis. AB - Anterior uveitis associated with Behcet's disease and ankylosing spondylitis preferentially occurs in adult men, which may suggest the effects of sex hormones on acute anterior uveitis. Recently, estrogen receptors in the vascular endothelium have been reported to be involved in several pathological conditions. In the present study, we examined the gender differences in susceptibility to endotoxin-induced uveitis (EIU) and the effects of estrogen on anterior inflammation. EIU was induced in adult male, female, and ovariectomized female Lewis rats (200 g) by hind footpad injection of 200 microg of LPS. In EIU, cellular infiltration was more marked in male than in female rats, and ovariectomy increased cellular infiltration. Treatment with 10 microg of 17beta estradiol significantly reduced the cell number in male and ovariectomized female rats with EIU. Estrogen receptor immunoreactivity was found in the nucleus of vascular endothelium and in some stromal cells of the iris-ciliary body. Semiquantitative PCR revealed that E-selectin and IL-6 gene expressions were increased in rats following LPS injection, and an overdose of tamoxifen, an estrogen receptor antagonist, reversed the effect of 17beta-estradiol on E selectin, but not its effect on IL-6. These observations suggested that the down modulation of these inflammatory genes by estrogen may contribute to the reduction in cellular infiltration in acute anterior uveitis. PMID- 10384137 TI - P-selectin binding promotes the adhesion of monocytes to VCAM-1 under flow conditions. AB - This study examined the adhesive interaction of peripheral blood monocytes with VCAM-1 and analyzed the effect of P-selectin binding to monocytes on the adhesive interaction with VCAM-1 under flow conditions. P-selectin glycoprotein ligand-1 is expressed on most monocytes. Furthermore, most monocytes bind soluble P selectin derived from platelets. P-selectin binding to monocytes did not alter the amount of expression of alpha4 integrin on monocytes. However, the mean channel fluorescence value for binding Cy2-conjugated soluble VCAM-1 to P selectin-bound monocytes was slightly more than that for binding Cy2-conjugated soluble VCAM-1 to untreated monocytes. Under flow conditions, the number of P selectin-bound monocytes bound to VCAM-1 was much higher than that of untreated monocytes bound to VCAM-1. These bindings were abolished by pretreatment of untreated monocytes and P-selectin-bound monocytes with anti-VCAM-1 mAb or anti alpha4 integrin mAb. Furthermore, P-selectin binding to monocytes increased shear resistance and thus increased the adhesive strength of monocytes to VCAM-1. These findings indicate that P-selectin binding to monocytes enhances the adhesive interaction of monocytes with VCAM-1. It is suggested that P-selectin glycoprotein ligand-1/P-selectin interaction and alpha4 integrin/VCAM-1 interaction can act sequentially in the adhesion cascade that regulates monocyte trafficking to inflammatory and atherosclerotic lesion. PMID- 10384140 TI - Pulmonary surfactant protein A modulates the cellular response to smooth and rough lipopolysaccharides by interaction with CD14. AB - Pulmonary surfactant protein A (SP-A) plays an important part in Ab-independent host defense mechanisms of the lung. In this study we investigated how SP-A interacts with distinct serotypes of bacterial LPS and modulates LPS-elicited cellular responses. SP-A bound to rough forms but not to smooth forms of LPS. In the macrophage-like cell line U937, SP-A inhibited mRNA expression and secretion of TNF-alpha induced by smooth LPS, but rough LPS-induced TNF-alpha expression was unaffected by SP-A. When U937 cells and rat alveolar macrophages were preincubated with SP-A, smooth LPS failed to induce TNF-alpha secretion, whereas rough LPS-induced TNF-alpha secretion was modestly increased. To clarify the mechanism by which SP-A modulates LPS-elicited cellular responses, we further examined the interaction of SP-A with CD14, which is known as a major LPS receptor. Western blot analysis revealed that CD14 was one of the SP-A binding proteins isolated from solubilized U937 cells. In addition, SP-A directly bound to recombinant soluble CD14 (rsCD14). When rsCD14 was preincubated with SP-A, the binding of rsCD14 to smooth LPS was significantly reduced but the association of rsCD14 with rough LPS was augmented. These results demonstrate the different actions of SP-A upon distinct serotypes of LPS and indicate that the direct interaction of SP-A with CD14 constitutes a likely mechanism by which SP-A modulates LPS-elicited cellular responses. PMID- 10384139 TI - Differential cytokine modulation and T cell activation by two distinct classes of thalidomide analogues that are potent inhibitors of TNF-alpha. AB - TNF-alpha mediates both protective and detrimental manifestations of the host immune response. Our previous work has shown thalidomide to be a relatively selective inhibitor of TNF-alpha production in vivo and in vitro. Additionally, we have recently reported that thalidomide exerts a costimulatory effect on T cell responses. To develop thalidomide analogues with increased anti-TNF-alpha activity and reduced or absent toxicities, novel TNF-alpha inhibitors were designed and synthesized. When a selected group of these compounds was examined for their immunomodulatory activities, different patterns of cytokine modulation were revealed. The tested compounds segregated into two distinct classes: one class of compounds, shown to be potent phosphodiesterase 4 inhibitors, inhibited TNF-alpha production, increased IL-10 production by LPS-induced PBMC, and had little effect on T cell activation; the other class of compounds, similar to thalidomide, were not phosphodiesterase 4 inhibitors and markedly stimulated T cell proliferation and IL-2 and IFN-gamma production. These compounds inhibited TNF-alpha, IL-1beta, and IL-6 and greatly increased IL-10 production by LPS induced PBMC. Similar to thalidomide, the effect of these agents on IL-12 production was dichotomous; IL-12 was inhibited when PBMC were stimulated with LPS but increased when cells were stimulated by cross-linking the TCR. The latter effect was associated with increased T cell CD40 ligand expression. The distinct immunomodulatory activities of these classes of thalidomide analogues may potentially allow them to be used in the clinic for the treatment of different immunopathological disorders. PMID- 10384141 TI - Decreased expression of Th2 type cytokine mRNA contributes to the lack of allergic bronchial inflammation in aged rats. AB - Sensitized Brown Norway rats are known to develop eosinophilic bronchial inflammation and airway hyperresponsiveness after Ag exposure. However, we have previously observed that sensitized aged rats of the same strain failed to develop such allergic inflammation. In the present study, we investigated age associated changes of cytokine mRNA expression in bronchoalveolar lavage (BAL) cells. Both young (8- to 10-wk-old) and aged (100- to 120-wk-old) Brown Norway rats were sensitized with OVA, and BAL was performed 24 h after OVA inhalation challenge. Semiquantitative RT-PCR analysis of BAL cells showed that the cells from aged rats preferentially expressed Th1 type cytokine (IFN-gamma) mRNA, whereas cells from young animals expressed more Th2 type cytokine mRNAs including those for IL-4 and IL-5. Decreased expression of Th2 type cytokine transcripts in aged animals was further confirmed by quantitative analysis, competitive RT-PCR of BAL cells, and in situ hybridization. The age-associated changes of cytokine profile were not restricted to BAL cells but were a general feature of lymphocytes, as shown by examination of popliteal lymph nodes draining the site of sensitization. These findings suggest that decreased allergic inflammation in aged animals is attributable to age-dependent impairment of Th2 generation in response to Ag. PMID- 10384142 TI - Mouse monocyte-derived chemokine is involved in airway hyperreactivity and lung inflammation. AB - The cloning, expression, and function of the murine (m) homologue of human (h) monocyte-derived chemokine (MDC) is reported here. Like hMDC, mMDC is able to elicit the chemotactic migration in vitro of activated lymphocytes and monocytes. Among activated lymphocytes, Th2 cells were induced to migrate most efficiently. mMDC mRNA and protein expression is modulated during the course of an allergic reaction in the lung. Neutralization of mMDC with specific Abs in a model of lung inflammation resulted in prevention of airway hyperreactivity and significant reduction of eosinophils in the lung interstitium but not in the airway lumen. These data suggest that mMDC is essential in the transit/retention of leukocytes in the lung tissue rather than in their extravasation from the blood vessel or during their transepithelial migration into the airways. These results also highlight the relevance of factors, such as mMDC, that regulate the migration and accumulation of leukocytes within the tissue during the development of the key physiological endpoint of asthma, airway hyperreactivity. PMID- 10384143 TI - The murine antiapoptotic protein A1 is induced in inflammatory macrophages and constitutively expressed in neutrophils. AB - Myeloid leukocytes are thought to regulate their susceptibility to apoptosis upon migration to a site of inflammation. However, factors that determine survival have not been well characterized in these cells. We have examined the expression of murine A1, an antiapoptotic Bcl-2 relative found in activated myeloid cells, during the course of an acute inflammatory response. Intraperitoneal infection of mice with the virulent RH strain of Toxoplasma gondii led to a 5- to 10-fold increase in A1 mRNA levels in peritoneal cells after several days. Bcl-2 expression was unchanged. The increase in A1 expression depended on the dose of the organism and coincided with a sharp increase in peritoneal cellularity. A1 protein levels were also increased as determined by Western blot analysis and immunohistochemical studies. All neutrophils and approximately half of the macrophages in the inflammatory exudate contained high levels of A1 in cytoplasm. A1 expression did not correlate with intracellular parasitization. Peripheral blood neutrophils from normal mice strongly expressed A1 protein, whereas normal monocytes showed only weak staining. Bax mRNA was induced in parallel with A1 in macrophages. Exudate macrophages and granulocytes that were apoptotic by TUNEL staining occasionally appeared to display A1 throughout the cell nucleus. These studies identify A1 as a potential regulator of apoptosis during acute inflammation. PMID- 10384144 TI - Induction of phosphorylation and intracellular association of CC chemokine receptor 5 and focal adhesion kinase in primary human CD4+ T cells by macrophage tropic HIV envelope. AB - Binding of HIV-1 envelope glycoproteins to the surface of a CD4+ cell transduces intracellular signals through the primary envelope receptor, CD4, and/or the envelope coreceptor, a seven-transmembrane chemokine receptor. Macrophage-tropic strains of HIV-1 preferentially use CCR5 as an entry coreceptor, whereas T cell tropic strains use CXC chemokine receptor-4 for entry. Intracellular signals transduced by HIV-1 envelope may have immunopathogenic consequences, including anergy, syncytium formation, apoptosis, and inappropriate cell trafficking. We demonstrate here that a recombinant envelope protein derived from an M-tropic isolate of HIV-1 can transduce CD4-dependent as well as CCR5-dependent intracellular signals in primary human CD4+ T cells. Novel HIV-induced intracellular signals that were identified include tyrosine phosphorylation of focal adhesion kinase (FAK) and CCR5, which are involved in cell adhesion and chemotaxis, respectively. HIV envelope-induced cellular association of FAK and CCR5 was also demonstrated, suggesting that ligation of CD4 and CCR5 leads to the formation of an activation complex composed of FAK and CCR5. Activation of this signaling pathway by HIV-1 envelope may be an important pathogenic mechanism of dysregulated cellular activation and trafficking during HIV infection. PMID- 10384145 TI - NF-kappa B regulation by I kappa B kinase in primary fibroblast-like synoviocytes. AB - NF-kappa B is a key regulator of inflammatory gene transcription and is activated in the rheumatoid arthritis (RA) synovium. In resting cells, NF-kappa B is retained as an inactive cytoplasmic complex by its inhibitor, I kappa B. Phosphorylation of I kappa B targets it for proteolytic degradation, thereby releasing NF-kappa B for nuclear translocation. Recently, two related I kappa B kinases (IKK-1 and IKK-2) were identified in immortalized cell lines that regulate NF-kappa B activation by initiating I kappa B degradation. To determine whether IKK regulates NF-kappa B in primary cells isolated from a site of human disease, we characterized IKK in cultured fibroblast-like synoviocytes (FLS) isolated from synovium of patients with RA or osteoarthritis. Immunoreactive IKK protein was found to be abundant in both RA and osteoarthritis FLS by Western blot analysis. Northern blot analysis showed that IKK-1 and IKK-2 genes were constitutively expressed in all FLS lines. IKK function in FLS extracts was determined by measuring phosphorylation of recombinant I kappa B in vitro. IKK activity in both RA and osteoarthritis FLS was strongly induced by TNF-alpha and IL-1 in a concentration-dependent manner. Activity was significantly increased within 10 min of stimulation and declined to near basal levels within 80 min. Activation of IKK in FLS was accompanied by phosphorylation and degradation of endogenous I kappa B alpha as determined by Western blot analysis. Concomitant activation and nuclear translocation of NF-kappa B was documented by EMSA and immunohistochemistry. Transfection with a dominant negative IKK-2 mutant prevented TNF-alpha-mediated NF-kappa B nuclear translocation, whereas a dominant negative IKK-1 mutant had no effect. This is the first demonstration that IKK-2 is a pivotal regulator of NF-kappa B in primary human cells. PMID- 10384146 TI - Osteoclast differentiation factor acts as a multifunctional regulator in murine osteoclast differentiation and function. AB - Osteoclast differentiation factor (ODF), a novel member of the TNF ligand family, is expressed as a membrane-associated protein by osteoblasts/stromal cells. The soluble form of ODF (sODF) induces the differentiation of osteoclast precursors into osteoclasts in the presence of M-CSF. Here, the effects of sODF on the survival, multinucleation, and pit-forming activity of murine osteoclasts were examined in comparison with those of M-CSF and IL-1. Osteoclast-like cells (OCLs) formed in cocultures of murine osteoblasts and bone marrow cells expressed mRNA of RANK (receptor activator of NF-kappaB), a receptor of ODF. The survival of OCLs was enhanced by the addition of each of sODF, M-CSF, and IL-1. sODF, as well as IL-1, activated NF-kappaB and c-Jun N-terminal protein kinase (JNK) in OCLs. Like M-CSF and IL-1, sODF stimulated the survival and multinucleation of prefusion osteoclasts (pOCs) isolated from the coculture. When pOCs were cultured on dentine slices, resorption pits were formed on the slices in the presence of either sODF or IL-1 but not in that of M-CSF. A soluble form of RANK as well as osteoprotegerin/osteoclastogenesis inhibitory factor, a decoy receptor of ODF, blocked OCL formation and prevented the survival, multinucleation, and pit forming activity of pOCs induced by sODF. These results suggest that ODF regulates not only osteoclast differentiation but also osteoclast function in mice through the receptor RANK. PMID- 10384147 TI - In situ T cell responses against melanoma comprise high numbers of locally expanded T cell clonotypes. AB - It is well established that melanoma cells express Ags that are recognized by autologous T cells in vitro. Tumor-infiltrating lymphocytes in situ comprise clonotypic T cells, suggesting that their expansion is driven by Ag stimulation. Still, little is known about the detailed characteristics of the in situ T cell response. In the present study, we scrutinized this response by analyzing multiple metastatic lesions for the presence of clonotypic T cells. This approach was chosen to distinguish whether the clonal T cell expansion occurs as a systemic or localized phenomenon. TCR clonotype mapping of six s.c. metastases from two patients revealed the presence of multiple (from 40 to >60) clonotypic T cells in all lesions. Clonotypic T cells were present in TCR beta-variable regions expressed both at high and low levels. Comparison of the T cell clonotypes present in different lesions from individual patients demonstrated that, in general, clonotypes were exclusively detected in a single lesion. Hence, anti-melanoma T cell responses are much more heterogeneous than previously anticipated and accommodate a predominance of strictly localized T cell clonotypes. PMID- 10384148 TI - Repeated administration of adenoviral vectors in lungs of human CD4 transgenic mice treated with a nondepleting CD4 antibody. AB - The central role of CD4+ T cells in regulation of adenovirus vector-mediated immune responses has been documented previously in murine models. We analyzed the effects of a nondepleting mAb to human CD4 (CD4 mAb; Clenoliximab) on immune functions following intratracheal administration of adenoviral vectors in murine CD4-deficient mice (muCD4KO) expressing a human CD4 transgene (HuCD4 mice). Treatment of HuCD4 mice with Clenoliximab inhibited both cell-mediated and humoral immune responses to adenoviral Ags. Chronic treatment of HuCD4 mice with Clenoliximab permitted successful readministration of adenoviral vectors at least four times. The ability to readminister these vectors is associated with marked suppression of neutralizing Ab responses to viral capsid proteins. Clenoliximab also inhibited CTL and prolonged expression of the transgene. T or B cell responses to adenovirus did not emerge after the effects of a short course of Clenoliximab diminished. These data illustrate the potential utility of a nondepleting CD4 Ab in facilitating gene therapy using adenoviral vectors. PMID- 10384149 TI - IL-5 increases expression of 5-lipoxygenase-activating protein and translocates 5 lipoxygenase to the nucleus in human blood eosinophils. AB - Cysteinyl-leukotrienes are potent bronchoconstrictor mediators synthesized by the 5-lipoxygenase (5-LO) pathway. Eosinophilopoietic cytokines such as IL-5 enhance cysteinyl-leukotriene synthesis in eosinophils in vitro, mimicking changes in eosinophils from asthmatic patients, but the mechanism is unknown. We hypothesized that IL-5 induces the expression of 5-LO and/or its activating protein FLAP in eosinophils, and that this might be modulated by anti inflammatory corticosteroids. Compared with control cultures, IL-5 increased the proportion of normal blood eosinophils immunostaining for FLAP (65 +/- 4 vs 34 +/ 4%; p < 0.0001), enhanced immunoblot levels of FLAP by 51 +/- 14% (p = 0.03), and quadrupled ionophore-stimulated leukotriene C4 synthesis from 5.7 to 20.8 ng/106 cells (p < 0.02). IL-5 effects persisted for 24 h and were abolished by cycloheximide and actinomycin D. The proportion of FLAP+ eosinophils was also increased by dexamethasone (p < 0.0001). Neither IL-5 nor dexamethasone altered 5 LO expression, but IL-5 significantly increased 5-LO immunofluorescence localizing to eosinophil nuclei. Compared with normal subjects, allergic asthmatic patients had a greater proportion of circulating FLAP+ eosinophils (46 +/- 6 vs 27 +/- 3%; p < 0.03) and a smaller IL-5-induced increase in FLAP immunoreactivity (p < 0.05). Thus, IL-5 increases FLAP expression and translocates 5-LO to the nucleus in normal blood eosinophils in vitro. This is associated with an enhanced capacity for cysteinyl-leukotriene synthesis and mimics in vivo increases in FLAP expression in eosinophils from allergic asthmatics. PMID- 10384151 TI - A complementary peptide vaccine that induces T cell anergy and prevents experimental allergic neuritis in Lewis rats. AB - We have developed and described a new method of altering T cell-mediated autoimmune diseases by immunization with the complementary peptide against T cell epitopes. The complementary peptide (denoted NAE 07-06) to the bovine P2 protein, residues 60-70 (denoted EAN 60-70), was tested in the Lewis rat model of experimental allergic neuritis (EAN). Immunization with NAE 07-06 induced polyclonal and monoclonal Abs that inhibited the proliferation of the P2-specific T cell line, stimulated with EAN 60-70, and recognized Vbeta, but not Valpha, of TCRs. Proliferation of T cells treated with anti-NAE 07-06 Abs could be partially restored by treatment with rIL-2, in accordance with an anergy model. A homologous sequence was found between NAE 07-06 and the VDJ junction of the TCR beta-chain from an EAN 60-70-specific T cell line. Rats preimmunized with NAE 07 06 in vivo before EAN induction showed less disease severity clinically and histologically. These data suggest a new therapeutic approach for T cell-mediated autoimmune disorders through the induction of anti-TCR Abs with complementary peptide Ags. PMID- 10384150 TI - Skin homing (cutaneous lymphocyte-associated antigen-positive) CD8+ T cells respond to superantigen and contribute to eosinophilia and IgE production in atopic dermatitis. AB - In allergic inflammations of the skin, activation of CD4+ T cells was demonstrated to play an important role; however, a minor role for CD8+ T cells is implied. In the present study, we compared cutaneous lymphocyte-associated Ag (CLA)-expressing CD4+ and CD8+ subsets, which were isolated from peripheral blood and lesional skin biopsies in atopic dermatitis (AD) patients. We demonstrated that CD8+CLA+ T cells proliferate in response to superantigen and are as potent as CD4+CLA+ T cells in IgE induction and support of eosinophil survival. In atopic skin inflammation, the existence of high numbers of CD4+ and CD8+ T cells was demonstrated by immunohistochemistry and by culturing T cells from skin biopsies. In peripheral blood, both CD4+ and CD8+ subsets of CLA+CD45RO+ T cells were in an activated state in AD. The in vivo-activated CLA+ T cells of both subsets spontaneously released an IL-5- and IL-13-dominated Th2 type cytokine pattern. This was confirmed by intracytoplasmic cytokine staining immediately after isolation of the cells from peripheral blood. In consequence, both CD4+ and CD8+, CLA+ memory/effector T cells induced IgE production by B cells mainly by IL 13, and enhanced eosinophil survival in vitro by delaying eosinophil apoptosis, mainly by IL-5. These results indicate that in addition to the CD4+ subset, the CD8+CLA+ memory/effector T cells are capable of responding to superantigenic stimulation and play an important role in the pathogenesis of AD. PMID- 10384152 TI - A non-AUG-defined alternative open reading frame of the intestinal carboxyl esterase mRNA generates an epitope recognized by renal cell carcinoma-reactive tumor-infiltrating lymphocytes in situ. AB - A number of Ags recognized by tumor-reactive T cells have been characterized, including nonmutated gene products and a variety of epitopes shown to arise from either mutated or alternatively processed transcripts. Here, we report that the screening of a cDNA library with an HLA-B7-restricted renal cell carcinoma reactive T cell clone derived from tumor-infiltrating lymphocytes (TILs) that were clonally amplified in vivo (as assessed by TCRBV complementarity determining region-3 length distribution analysis) resulted in the isolation of a nonamer encoded by an alternative open reading frame (ORF) (a +1 frameshift) of the intestinal carboxyl esterase gene. This peptide binds HLA-B*0702-presenting molecules as assessed in an immunofluorescence-based peptide binding assay using transfected T2 cells. Constitutive expression of this alternative ORF protein was observed in all transformed HLA-B7+ renal cell lines that were recognized in cytotoxicity assays by the TILs. The intestinal carboxyl esterase gene is transcribed in renal cell carcinoma tumors as well as in normal liver, intestinal, or renal tissues. Mutation of the natural ATG translation initiation site did not alter recognition, indicating that frameshifting (i.e., slippage of the ribosome forward) and recoding are not involved. In addition, a point mutation of the three AUG codons that may be used as alternative translation initiation sites in the +1 ORF did not abolish recognition, whereas mutation of an upstream ACG codon did, indicating that the latter codon initiates the translation of the alternative ORF. These results further extend the types of Ags that can be recognized by tumor-reactive TILs in situ (i.e., leading to clonal T cell expansion). PMID- 10384153 TI - Altered memory T cell differentiation in patients with early rheumatoid arthritis. AB - The chronic immune response in rheumatoid arthritis (RA) might be driven by activated Th1 cells without sufficient Th2 cell differentiation to down-modulate inflammation. To test whether disordered memory T cell differentiation contributes to the typical Th1-dominated chronic inflammation in RA we investigated differentiation of resting CD4+ memory T cells in patients with early (6 wk to 12 mo) untreated RA and in age- and sex-matched healthy controls in vitro. No difference in cytokine secretion profiles of freshly isolated memory T cells was detected between patients and controls. A cell culture system was then employed that permitted the differentiation of Th effectors from resting memory T cells by short term priming. Marked differences were found in response to priming. Th2 cells could be induced in all healthy controls by priming with anti-CD28 in the absence of TCR ligation. By contrast, priming under those conditions resulted in Th2 differentiation in only 9 of 24 RA patients. Exogenous IL-4 could overcome the apparent Th2 differentiation defect in seven patients but was without effect in the remaining eight patients. In all patients a marked decrease in IL-2-producing cells and a significant increase in well differentiated Th1 cells that produced IFN-gamma but not IL-2 were evident after priming with anti-CD3 and anti-CD28. The data suggest that CD4+ memory T cells from patients with early untreated RA manifest an intrinsic abnormality in their ability to differentiate into specific cytokine-producing effector cells that might contribute to the characteristic Th1-dominated chronic (auto)immune inflammation in RA. PMID- 10384154 TI - Endogenous CD8+ T cell expansion during regression of monoclonal EBV-associated posttransplant lymphoproliferative disorder. AB - There are experimental data which suggest that the primary immune effector cell responsible for maintaining immune surveillance against the outgrowth of EBV transformed B cells in humans is the CTL, but in vivo proof of this is lacking. In this study we perform a series of cellular and molecular assays to characterize an autologous, endogenous immune response against a transplantation associated, monoclonal, EBV+ posttransplant lymphoproliferative disorder (PTLD). Following allogeneic bone marrow transplantation, a patient developed a monoclonal PTLD of donor B cell origin. With a decrease in immune suppression, we document the emergence of endogenous, donor-derived CD3+CD8+ CTLs, followed by regression of the PTLD. The TCR Vbeta repertoire went from a polyclonal pattern prior to the development of PTLD to a restricted TCR Vbeta pattern during the outgrowth and regression of PTLD. Donor-derived CD3+CD8+ T lymphocytes displayed MHC class I-restricted cytolytic activity against the autologous EBV+ B cells ex vivo without additional in vitro sensitization. The striking temporal relationship between the endogenous expansion of a TCR Vbeta-restricted, CD3+CD8+ population of MHC class I-restricted CTL, and the regression of an autologous monoclonal PTLD, provides direct evidence in humans that endogenous CD3+CD8+ CTLs can be responsible for effective immune surveillance against malignant transformation of EBV+ B cells. PMID- 10384155 TI - Efficient transfer of a tumor antigen-reactive TCR to human peripheral blood lymphocytes confers anti-tumor reactivity. AB - The tumor-associated-Ag MART-1 is expressed by most human melanomas. The genes encoding an alphabeta TCR from a MART-1-specific, HLA-A2-restricted, human T cell clone have been efficiently transferred and expressed in human PBL. These retrovirally transduced PBL cultures were MART-1 peptide reactive, and most cultures recognized HLA-A2+ melanoma lines. Limiting dilution clones were generated from three bulk transduced PBL cultures to investigate the function of individual clones within the transduced cultures. Twenty-nine of 29 CD8+ clones specifically secreted IFN-gamma in response to T2 cells pulsed with MART-1(27-35) peptide, and 23 of 29 specifically secreted IFN-gamma in response to HLA-A2+ melanoma lines. Additionally, 23 of 29 CD8+ clones lysed T2 cells pulsed with the MART-1(27-35) peptide and 15 of 29 lysed the HLA-A2+ melanoma line 888. CD4+ clones specifically secreted IFN-gamma in response to T2 cells pulsed with the MART-1(27-35) peptide. TCR gene transfer to patient PBL can produce CTL with anti tumor reactivity in vitro and could potentially offer a treatment for patients with metastatic melanoma. This approach could also be applied to the treatment of other tumors and viral infections. Additionally, TCR gene transfer offers unique opportunities to study the fate of adoptively transferred T cells in vivo. PMID- 10384156 TI - Induced expression of B7-1 on myeloma cells following retroviral gene transfer results in tumor-specific recognition by cytotoxic T cells. AB - The aim of this study was to evaluate whether tumor cells from patients with multiple myeloma activate allogeneic and autologous T cells. Results showed that myeloma cells expressed few B7-2 and no B7-1 in six cell lines and primary cells from 11 patients. They expressed substantial levels of HLA class I, CD40, and a set of adhesion molecules. In accordance with the low density of B7 molecules on these cells, they were poor allogeneic CD8+ T cell stimulators. Neither IFN-gamma plus TNF-alpha nor CD40 stimulation significantly induced B7-1 or up-regulated B7 2 on human myeloma cell line or primary myeloma cells from six of seven patients. However, such induction was found on autologous bone-marrow nontumoral cells and on autologous dendritic cells following CD40 stimulation. High B7-1 expression was stably obtained on human myeloma cell line using transduction with a B7-1 retrovirus, enabling these cells to stimulate allogeneic CD8+, though not CD4+, T cell proliferation. For one patient with advanced disease, B7-1 gene transfer made it possible to amplify autologous cytotoxic T cells that killed autologous myeloma cells in an HLA class I-restricted manner, but not autologous PHA blasts. These results suggest that B7-1 gene transfer could be a promising immunotherapeutic approach in multiple myeloma. PMID- 10384157 TI - Early autoantibody responses in prediabetes are IgG1 dominated and suggest antigen-specific regulation. AB - The islet autoimmunity of preclinical type 1 diabetes remains poorly characterized in humans. In this paper, the IgG subclass response to the islet autoantigens insulin, glutamic acid decarboxylase, and IA-2 was studied sequentially from birth to diabetes onset or current follow-up in 26 autoantibody positive offspring of parents with diabetes. Islet autoantibody appearance was characterized by an early IgG1 peak response to one or more Ags, most commonly to insulin, at a median age of 2.2 yr (interquartile range, 2-2.9 yr). In five offspring, an acute fulminant beta-cell destruction and diabetes onset occurred during this initial Ab response. In the remainder, early Ab levels declined markedly, and Ab peaks against other beta cell Ags arose sequentially over several years suggesting regulation and spreading of autoimmunity. Second peak Ab responses to the same Ag were observed in only two offspring, both developing diabetes at this time. Two others developed diabetes with declining Ab levels. Abs of IgG1 subclass dominated against each Ag, and other subclasses, were usually only detected during peak IgG1 responses. The IgG4 response to insulin was exceptional, being dominant over IgG1 in four offspring and in five others appeared and/or persisted after IgG1 levels declined. These Th2-associated IgG4 responses were not correlated with protection from diabetes. The presence of IgG1 restricted responses to DA2 were associated with diabetes development. These findings suggest that type 1 diabetes has an early acute destructive phase of beta cell autoimmunity, which may be regulated and which spreads chronically until diabetes onset. PMID- 10384158 TI - Increased apoptosis in patients with major depression: A preliminary study. AB - Apoptosis is a programmed cell death that can be observed in normal cells. Major depression poses a combination of a depressed and destructive autoimmune reaction. We measured apoptosis in the PBLs of seven patients with major depression and in age- and sex-matched controls. We observed significantly increased apoptosis in the PBLs of depressive patients (p < 0.05). These preliminary results could contribute to an understanding of the interactions of the CNS with the immune system, which could lead to the increased vulnerability of the CNS in depressive disorders. Further studies are needed to establish these results. PMID- 10384159 TI - Diagnosis and management of recurrent herpes simplex induced by fixed prosthodontic tissue management: a clinical report. PMID- 10384160 TI - 1998 Judson C. Hickey Scientific Writing Award. Maxillofacial prosthodontic management of a facial defect complicated by a necrotic frontal bone flap: a clinical report. PMID- 10384161 TI - Qualitative and quantitative determination of dental amalgam restoration volume. AB - STATEMENT OF PROBLEM: Volume of tooth structure replaced by an existing restoration, as assessed by visual and radiographic examination, is one diagnostic measure used by dental practitioners and dental insurance agencies to determine the relative need to restore a tooth with a full-coverage cast restoration. However, use of these methods has not been validated. PURPOSE: This study compared the volume of a range of dental amalgam restorations placed in typodont teeth, as estimated by dentists, dental students and laypersons, with the actual volume of each restoration. METHODS AND MATERIAL: Sixty subjects (20 dental school faculty, 20 dental students, and 20 clerical staff [laypersons]) participated. After reviewing photographic images of typodont teeth with mesial occlusal-distal dental amalgam restorations, subjects estimated the volume of each restoration using various restorations on different teeth as a percentage of its tooth's coronal volume. The actual volume of each dental amalgam restoration and that of the coronal portion of the prepared teeth was calculated with a volumetric displacement technique. The single sample 2-sided t test with a.05 level of significance was used to evaluate the null hypothesis (H0 ): The survey participant's estimates of each restoration's percentage volume are the same as the measured volume values versus the alternative hypothesis (H1 ): estimates differed from the measured volume values. One-way analysis of variance was used to determine the significance of any difference between the estimates of the 3 survey test groups. RESULTS: Average volumes reported by all 3 groups were significantly different than the measured volume values (P <.05). Experience and dental training did not significantly affect a participant's ability to evaluate restoration volumes with greater accuracy. Results reported by dentists, dental students, and laypersons were not significantly different (P >.05). CONCLUSIONS: The volume of a restoration is inaccurately assessed by visual and radiographic examination. PMID- 10384162 TI - Investigation of the effect of three sprue designs on the porosity and the completeness of titanium cast removable partial denture frameworks. AB - STATEMENT OF PROBLEM: Although titanium has been used to cast removable partial denture frameworks, the casting process is arduous and requires specialized equipment. PURPOSE: This study evaluated the ability of 3 sprue designs (tree, ball, and circular) to produce complete, void-free castings of removable partial denture frameworks made from commercially pure titanium. METHODS AND MATERIAL: A cast with a Kennedy class III, modification 1, partially edentulous arch was used. The blocked-out cast was modified to facilitate assessment of the completeness of the casting of the clasp arms. Thirty refractory casts were made, and 10 wax patterns for each sprue design were fabricated and invested with ethyl silicate investment. Castings were made with an arc-type automatic casting machine in an argon atmosphere. Castings were examined with radiographic equipment to detect the presence of voids in the castings, and the completeness to each casting was visually verified. Data on porosities were tabulated and statistically analyzed with 1-way ANOVA followed by Student Newman-Keuls test. Chi-square analysis was used to identify statistically significant differences in casting defects among the 3 sprue designs. RESULTS: No statistically significant differences in the total number of porosities were found between the 3 sprue designs (P =.51). Results of completeness of castings evaluated by using chi square test revealed a statistically significant deference among the 3 sprue designs (P =.008). CONCLUSION: The ball-sprue design produced the most complete castings for the removable partial denture titanium frameworks. PMID- 10384163 TI - Effect of a prefabricated anterior bite stop on electromyographic activity of masticatory muscles. AB - STATEMENT OF PROBLEM: Patients are often treated for signs and symptoms in the masticatory musculature, which may be manifested as pain and/or conditions that cause difficulty in recording jaw relation records. A quick, easy method to alleviate these signs and symptoms would be helpful. PURPOSE: This study measured the effect of a prefabricated anterior bite stop on the electromyographic activity of the anterior temporalis, posterior temporalis, masseter and anterior digastric during clenching, and grinding tasks. MATERIAL AND METHODS: A prefabricated anterior bite stop was fabricated for 30 randomly selected subjects. Electromyographic surface electrodes were placed on the right and left sides of these muscles. Electromyographic activity was measured during clenching and grinding both with and without the anterior bite stop. RESULTS: The anterior bite stop had a significant effect in decreasing electromyographic activity for both clenching and grinding for all the tested muscles, except the anterior digastric. CONCLUSIONS: For this patient population, the ready-made anterior bite stop reduced electromyographic muscle activity for the anterior and posterior temporalis and the masseter muscles during both clenching and grinding. PMID- 10384164 TI - Annual review of selected dental literature: report of the Committee on Scientific Investigation of the American Academy of Restorative Dentistry. PMID- 10384165 TI - Influence of mandibular superstructure shape on implant stresses during simulated posterior biting. AB - STATEMENT OF PROBLEM: Theoretical considerations on the ideal implant-supported prosthetic superstructure shape lack the effect of complex mandibular deformation patterns during function. PURPOSE: This study compared implant abutment stresses for idealized superstructures with different cross-sectional shapes and material properties during a simulated, complex biting task. MATERIAL AND METHODS: A simplified and idealized 3-dimensional finite element computer model was built, which consisted of a sectioned mandible rehabilitated with 5 titanium implants and an attached superstructure beam composed of metal alloy and acrylic resin. The model was submitted to loads mimicking simultaneous bending and (to a lesser degree) torsion of the mandibular corpus during a bilateral posterior bite. Maximum and minimum principal stresses were calculated at implant abutment sites for each of 6 beam cross sections of the prosthetic superstructure and 2 types of materials. RESULTS: Predicted implant stresses varied significantly between implant sites for different superstructure shapes. The lowest principal stresses were obtained by using a superstructure with a rectangular-shaped beam oriented vertically. Contrary to former theoretical considerations, the ideal "I-beam" superstructure cross section did not yield the lowest stresses. Superstructure materials with a lower modulus of elasticity seem to not only increase implant abutment stresses overall but also slightly reduce the tensile stresses on the most anterior implants. CONCLUSION: Simulated implant abutment stresses may be significantly affected by the shape of the prosthetic superstructure, by diverse mandibular loading conditions, and to a lesser extent, by the prosthetic material properties. PMID- 10384166 TI - Dental diamond burs made with a new technology. AB - STATEMENT OF PROBLEM: Conventional diamond burs show several limitations such as the heterogeneity of grain shapes, the difficulty of automation during fabrication, the decrease of cutting effectiveness due to repeated sterilization, and short lifetime. An additional shortcoming may be represented by the potential release of Ni+2 ions from the metallic binder into the body fluids. PURPOSE: This study investigated a new diamond rotative instrument made of a continuous diamond film obtained by chemical vapor deposition (CVD). This bur, characterized by a pure diamond cutting surface without metallic binder between crystals, was compared with a conventional diamond bur. MATERIAL AND METHODS: Cutting tests were followed by SEM examination and electron microprobe analysis (EMA) to trace metallic residues both at the surface of the bur and the substrate. RESULTS: EMA demonstrated that the metals Ni, Cr, Si, and Fe were present in the metallic binder matrix of the conventional bur and could be smeared on the surface of the substrate during cutting. SEM showed that significant loss of diamond particles occurred during cutting. On the other hand, no discrete particles sheared off the CVD bur. The smearing of the metallic binder cannot occur using the new bur. CONCLUSION: The new CVD bur not only proves to be more efficient in its cutting ability and longevity, but also excludes the risk of metal contamination. This last aspect concerns both the pollution of the oral environment and the contamination of the ceramic during the laboratory manufacturing of dental restorations. PMID- 10384167 TI - Professional attitudes toward denture adhesives: A Delphi technique survey of academic prosthodontists. AB - STATEMENT OF PROBLEM: The use of denture adhesives and their role in prosthodontics has been an intriguing and conflicted topic, both in clinical practice and dental education. PURPOSE: This study generated discussion, and if possible, obtained a consensus on a series of issues related to denture adhesives among a group of leading academic prosthodontists. MATERIAL AND METHODS: The Delphi Technique survey method was used. It consists of a series of survey questionnaire rounds to a panel of experts to either develop a consensus (>70% agreement) or to clarify the reasons for multiple schools of thought on a topic. A 24-item Delphi questionnaire was sent to an expert panel that consisted of a 33% regionally stratified random sample of program directors of undergraduate complete denture courses in US dental schools. The 5 major topic areas on denture adhesives addressed by the questionnaire items were (1) general perceptions, (2) specific clinical uses/misuses, (3) patient education, (4) inclusion in dental curricula, and (5) overall opinions of utility. RESULTS: Of the 18 randomly selected panelists, 94% (n = 17) agreed to participate, with either 16 or 17 fully participating in each of the 3 survey rounds. The panel achieved consensus and clearly concluded that denture adhesives: (1) are a useful adjunct in denture prosthesis services, having specific roles in both the fabrication and postinsertion phases; (2) had the potential for misuse, both by dentists and by patients; and (3) should be an integral part of patient education for all denture patients and of the undergraduate dental curriculum. However, the panel was unable to achieve consensus on whether denture adhesives should be used at the postinsertion phase for immediate denture patients and whether, on the whole, they were a beneficial adjunct in denture patient management (59% agreed they were). The panelists also clearly expressed their concerns that neither dentists nor patients should use denture adhesives as a substitute for either good clinical practices or proper denture maintenance routines. CONCLUSIONS: This panel of leading academic prosthodontists concluded that denture adhesives are a useful adjunct in denture prosthesis services, with specific roles in both fabrication and postinsertion phases. They also indicated that only through education, for dentists and patients, would the dual goals of maximizing the beneficial aspects of denture adhesive use while minimizing the misuse of denture adhesives be achieved. PMID- 10384168 TI - 1998 Judson C. Hickey Scientific Writing Award. Use of a flexible cast for the indirect fabrication of provisional restorations. AB - Efficient fabrication of a clinically acceptable provisional restoration for a fixed partial denture is an important part of treatment success. Fabrication of provisional restorations that uses the indirect technique produces accurate fitting provisional restorations without the chemical and thermal irritation associated with direct fabrication. With a typodont model, an indirect method is presented that uses an elastic cast for fabrication of multiple unit provisional restorations for fixed partial dentures. The cast is available within 6 minutes of impression making, can be trimmed with a sharp scalpel, and provides flexibility that allows easy separation of the acrylic provisional from the cast. The cast can also be used to evaluate the clinical acceptability of the preparations before impression making. This method has also been successfully used for the fabrication of acrylic provisional restorations for onlay preparations. PMID- 10384169 TI - Alternative complete-arch cement-retained implant-supported fixed partial denture. AB - Early implant prostheses designs, which used screw-retained metal and acrylic resin structures, frequently left a space between the prosthesis and the soft tissue. Common deficiencies of this design included phonetic and esthetic problems and screw loosening. Cement-retained implant prostheses are also used in partially and completely edentulous patients, and are thought to have optimal occlusion and esthetics. Moreover, cement-retained prostheses may induce less stress on the implant, thereby maximizing the possibility of a passive fit. Porcelain fused to metal prostheses have been used mostly in partially edentulous situations. Recently, complete-arch porcelain fused to metal prostheses that replace hard and soft tissue have been used and, although this restoration can have excellent esthetics, there are disadvantages such as high cost, potential framework distortion during fabrication, and difficulty in repairing fractures of in-service porcelain. This article describes an alternative technique for the fabrication of a complete-arch, cement-retained, metal-acrylic resin implant supported fixed partial denture. When compared with porcelain fused to metal complete-arch restorations, this prostheses is esthetic, has excellent retention and stability, yet is relatively inexpensive to fabricate, and requires less laboratory skill. PMID- 10384170 TI - Blockout technique for impressions of teeth with increased open gingival embrasures. AB - A dental technique is described that blocks out enlarged gingival embrasures to eliminate distortion of an impression and the resultant cast for removable partial or fixed partial dentures. This procedure consists of injecting polyvinyl siloxane impression material in embrasures to form custom blockout wedges. This technique provides a simple method for clean, customized blockout of potentially damaging undercuts that can distort impressions and casts. The ultimate accuracy of the cast results in a precise definitive prosthesis. PMID- 10384171 TI - Hinged mandibular removable complete denture for post-mandibulectomy patients. AB - The presence of excessive lingual undercuts after mandibulectomy and surgical reconstruction is a rare clinical condition and presents unique prosthodontic challenges. The goals of prosthodontic treatment include: providing lip support, improving articulation, reducing drooling, and regaining favorable esthetics. This article describes the fabrication of a hinged removable mandibular complete denture prosthesis using a sectional impression tray technique and a custom-made hinge mechanism. Clinical and laboratory procedures of the prosthetic treatment are described, and the advantages and disadvantages are reviewed. PMID- 10384172 TI - Functional criteria for mandibular implant placement post resection and reconstruction for cancer. AB - STATEMENT OF PROBLEM: Osseointegrated implants used in the mandible post resection and reconstruction for cancer represents a treatment option with the potential for functional improvement and enhanced quality of life. Unfortunately, protocols for their use in this patient population have been empirical and technique-driven with the assumption that they will overcome most, if not all, functional deficits encountered. PURPOSE: The article reviews the salient oral physiologic factors for this group of patients and presents a rational approach and functional criteria for patient selection and implant placement. Other considerations discussed include: timing of implant placement, irradiated and compromised tissues, patient motivation, and tumor prognosis. CONCLUSION: These principles, if followed, may enhance realistic functional outcomes for this patient population. PMID- 10384173 TI - Simplified method for fabrication of a claspless removable partial denture using extracoronal resilient attachment. PMID- 10384174 TI - Occlusal guard for the maxillary edentulous patient. PMID- 10384175 TI - Readers round table-AES/Alliance. American Equilibration Society/Alliance of TMD Practitioners. PMID- 10384176 TI - Endoscopic robotic coronary surgery is this reality or fantasy? PMID- 10384177 TI - Endoscopic coronary artery bypass grafting with the aid of robotic assisted instruments. AB - OBJECTIVE: The development of endoscopic coronary artery bypass grafting has been limited because of poor visualization and increased technical difficulties in carrying out operations through ports. We investigated whether the use of robotic assisted instruments could minimize these difficulties. METHODS: After a period of technical development and training on cadavers (n = 8) with the Intuitive Surgical system (Intuitive Surgical, Inc, Mountain View, Calif), the first clinical application in coronary artery surgery was performed in 4 male patients (mean age 59 +/- 6 years) with the indication of grafting the left internal thoracic artery to the left anterior descending coronary artery. Robotic assisted 3-dimensional endoscopes and instruments were introduced into the left side of the chest through 3 intercostal ports. The Heartport system (Heartport, Inc, Redwood City, Calif) was used for arresting the heart during the anastomosis. RESULTS: In 2 patients, the harvesting of the left internal thoracic artery was completed endoscopically with robotic assisted instruments and the anastomosis to the left anterior descending artery was performed through a minithoracotomy with conventional instruments. In 2 other patients, the entire operation was completed endoscopically with robotic assisted instruments. Early postoperative coronary angiography demonstrated the patency of the grafts in all cases. At 6-month follow-up, all patients were free of symptoms. CONCLUSIONS: Robotic assisted instruments make endoscopic coronary bypass possible and open a new era in minimally invasive surgery. PMID- 10384178 TI - Use of the voice-controlled and computer-assisted surgical system ZEUS for endoscopic coronary artery bypass grafting. AB - OBJECTIVE: With the aim of performing a completely endoscopic coronary bypass anastomosis, we have undertaken an experimental and clinical study using robotic instrumentation and voice-controlled camera guidance. METHODS: The ZEUS Robotic Surgical System (Computer Motion Inc, Goleta, Calif) consists of three interactive robotic arms and a control unit, allowing the surgeon to move the instrument arms in a scaled down mode. The third arm (AESOP, Computer Motion) positions the endoscope via voice control. PHASE I: In a phantom model, vascular grafts were anastomosed to the left anterior descending coronary artery (LAD) of 50 pig hearts with either 2- or 3-dimensional visualization. PHASE II: In 6 dogs (FBI 20-25 kg) the left internal thoracic artery (LITA) was harvested endoscopically. Then the animals were placed on an endovascular cardiopulmonary bypass system (Port-Access, Heartport, Inc, Redwood City, Calif). Anastomosis of the LITA to the LAD was performed endoscopically with the telemetric ZEUS instruments. Flow rates through the LITA were measured by Doppler analysis. PHASE III: Two patients were operated on with the ZEUS system. After endoscopic harvesting of the LITA and cardiopulmonary bypass with the Port-Access system, the bypass graft (LITA-LAD) was anastomosed endoscopically with the ZEUS system through three thoracic ports. RESULTS: In the dry laboratory, the time range required for the robotically assisted coronary anastomosis was 35 to 60 minutes with 2-dimensional visualization and 16 to 32 minutes with 3-dimensional visualization. In the animal experiments, the median time for endoscopic harvesting of the LITA was 86 minutes (range 56-120 minutes) and for the anastomosis, 42 minutes (range 35-105 minutes); flow rates through the LITA ranged between 22 and 45 mL/min. In the clinical cases, preparation times for the LITA were 83 and 110 minutes, respectively, and anastomosis times, 42 and 40 minutes, respectively. Doppler flow rates measured 125 and 85 mL/min, respectively. Both patients had an uneventful follow-up angiogram and postoperative course. CONCLUSIONS: With sophisticated robotic technology, a completely endoscopic anastomosis of the LITA to the LAD is possible, allowing technically precise operations within acceptable time limits. PMID- 10384180 TI - Commentary PMID- 10384179 TI - Preservation of intercostal arteries during thoracoabdominal aortic aneurysm surgery: a retrospective study. AB - OBJECTIVE: The purpose of this article was to examine the influence of reimplantation of patent intercostal and lumbar arteries on the incidence of postoperative paraplegia/paraparesis in patients undergoing clamp-and-sew surgical repair of thoracoabdominal aortic aneurysms. METHODS: Data from January 1987 through December 1997 were retrospectively collected on 132 patients. Ninety one patients in group I underwent aneurysm repairs before January 1995 and did not undergo intercostal artery reimplantation. Group II included the more recent 41 patients who had vessels between the eighth thoracic intercostal and the second lumbar arteries reimplanted to the graft or preserved at the aortic anastomoses. RESULTS: The operative mortality rate was 13.2% (12/91) in group I and 4.9% (2/41) in group II (P =.22). The incidence of postoperative paraplegia was significantly lower in the more recent cohort of patients (8.8% [8/91] in group I vs 0% [0/41] in group II, P =.05). The overall rate of spinal cord dysfunction was lowered from 9.9% (9/91) in group I to 2.4% (1/41) in group II (P =.17). However, a multivariable logistic regression analysis identified only aneurysm extent (Crawford type I and type II) as a predictor of less postoperative spinal cord injury (P =.08). The average aortic crossclamp time in group I was 30.3 +/- 11.5 (SD) minutes, and the time of aortic occlusion in group II was not significantly prolonged, with an average crossclamp time of 31.0 +/- 21.0 (SD) minutes (P =. 88). CONCLUSIONS: An aggressive approach to maintain intercostal artery patency during clamp-and-sew repair of thoracoabdominal aortic aneurysms may effectively lower the incidence of spinal cord injury without prolonging aortic crossclamp time. PMID- 10384181 TI - Exogenous control of cardiac gene therapy: evidence of regulated myocardial transgene expression after adenovirus and adeno-associated virus transfer of expression cassettes containing corticosteroid response element promoters. AB - OBJECTIVE: Because of the relative inaccessibility of the heart for repeated gene therapy, it would be useful to regulate the expression of transgenes delivered in a single dose of a gene therapy vector. Incorporation into the vector of a regulatable promoter that is responsive to pharmacologic agents that are widely used and well tolerated in clinical practice represents such a control strategy. METHODS: A replication-deficient adenovirus or an adeno-associated virus containing a chimeric promoter composed of 5 glucocorticoid response elements and the murine thrombopoietin complementary DNA (AdGRE.mTPO or AAVGRE.mTPO) was administered to the hearts of Sprague-Dawley rats. Platelet levels were evaluated as a reporter of transgene activity with or without dexamethasone. For comparison, rats received a control adenovirus vector, AdCMV.mTPO or AdCMV.Null, and the control adeno-associated virus vector AAVCMV.luc, which encodes for the firefly luciferase (luc) gene. RESULTS: Platelet elevation in the AdGRE.mTPO group peaked 4 days after dexamethasone administration, with a return to baseline 1 week after the initial corticosteroid dose. Subsequent dexamethasone administration at 2 and 4 weeks resulted in similar but progressively decreased responses. The AAVGRE.mTPO group had 5 peak platelet levels to a minimum of 2.2 fold with respect to baseline without diminution with subsequent dexamethasone administrations out to 169 days. In contrast, the AdCMV.Null and AAVCMV.luc groups demonstrated no increase in platelet counts and the AdCMV.mTPO group demonstrated a slow rise to a single peak platelet count independent of dexamethasone administration. CONCLUSION: It may be possible to control on demand the expression of a gene transferred to the heart. This strategy should be useful in cardiac gene therapy. PMID- 10384182 TI - Commentary PMID- 10384184 TI - Coronary artery bypass without cardiopulmonary bypass for patients with acute myocardial infarction. AB - OBJECTIVE: Between January 1992 and December 1994, 57 patients having an acute myocardial infarction with coronary anatomy suitable for coronary artery bypass grafting without cardiopulmonary bypass underwent this procedure within 1 week of the infarction. We describe the surgical results of these high-risk patients. METHODS: The study population included 43 male patients (75%) and 14 female patients (25%) whose mean age was 58.5 +/- 10.4 years. Thirty-two patients (56%) underwent emergency bypass grafting within 48 hours of an acute myocardial infarction, 4 of them (12.5%) as a bailout procedure after complicated percutaneous transluminal coronary angioplasty. Of these 32 patients, 7 patients (22%) were in cardiogenic shock, and 10 patients (31%) required preoperative intra-aortic balloon pump. Twenty-five patients (44%) underwent coronary bypass grafting 2 to 7 days after an acute myocardial infarction. The mean number of grafts per patient was 1.8 (range, 1-4), and the internal thoracic artery was used in 47 patients (82%). Only 7 patients (12%) received grafts to a circumflex marginal branch. RESULTS: Operative mortality was 1.7% (1 patient), and the mean postoperative hospital stay was 6.8 +/- 3 days. One- and 5-year actuarial survivals were 94.7% and 82.3%, respectively. Angina returned in 7 patients (12%), 1 of whom underwent reoperation. Multivariate analysis revealed renal failure and preoperative cardiogenic shock to be independent predictors of overall mortality. Old myocardial infarction and operation within the first 48 hours were independent predictors of overall unfavorable outcome events. CONCLUSIONS: These results suggest that coronary artery bypass grafting without cardiopulmonary bypass is a relatively low-risk procedure for patients having an infarction with coronary anatomy suitable for this technique. PMID- 10384183 TI - Interstitial cellular and matrix restoration of cardiac valves after cryopreservation. AB - OBJECTIVES: We previously characterized the porcine aortic leaflet interstitial cell phenotype as having both synthetic and contractile characteristics; that is, it is a myofibroblast. In this study we hypothesized (1) that the cryopreservation of aortic valves causes a significant reduction in cell density, (2) that it simultaneously causes alterations in representative components of extracellular matrix, and (3) that both of these processes are reversible. METHODS: Seventy-two leaflets from 24 porcine aortic valves were studied. Whole valves were subjected to variable lengths of preharvest ischemia (group 1), ischemia followed by processing analogous to clinical methods (group 2), and ischemia followed processing with an organ culture type of resuscitation (group 3). Vital dye exclusion by cells enzymatically dispersed from leaflets was used to quantify viability. Electron and light microscopy, immunohistochemical assay, and a silicone rubber substratum contractility assay were used both in dispersed cell preparations and in leaflet cross sections to examine structural, ultrastructural, and functional changes across the 3 groups through a range of preharvest ischemic times. RESULTS: Results indicated that harvest ischemic periods between 2 and 24 hours after donor death were not responsible for cell number reductions. During this interval overt dissolution of chondroitin sulfate simultaneous with a relative sparing of fibronectin was evidenced by immunohistochemical staining. Although not reduced in number, ischemic interstitial cells did show significant ultrastructural evidence of injury and suppressed monoclonal binding to vimentin and alpha-smooth muscle actin. After cryopreservation, viable cell numbers were always markedly reduced at all ischemic intervals and damage to both soluble extracellular matrix components and cell ultrastructure was increased. At all time and processing points, however, some retention of matrix secretory and cellular contractile capabilities was observed among the surviving cells. After the extended periods of preharvest ischemia (2-24 hours) followed by processing, a restitution of functioning cells was accomplished by means of whole-leaflet incubation in 15% fetal bovine serum. CONCLUSIONS: After application of the described methods, new cells within restored intact leaflets as well as in single-cell preparations demonstrated normal ultrastructure and contractile and synthetic functions (normal phenotypic expression). If functioning leaflet interstitial cells can contribute to homograft durability, bioengineering methods for pretransplantation cell repopulation could be refined with these techniques and applied to clinical valve transplantation. PMID- 10384185 TI - Influence of aortic valve replacement, prosthesis type, and size on functional outcome and ventricular mass in patients with aortic stenosis. AB - OBJECTIVES: Two years after surgery for severe aortic stenosis, we prospectively evaluated the influence of aortic valve replacement, as well as valve type (mechanical or stented biologic) and size, on functional status, left ventricular function, and regression of mass. METHODS: Patients who received either a mechanical (n = 95) or a biologic valve (n = 42) were studied by echocardiography before the operation and after 2 years. RESULTS: The percentage of patients with severe dyspnea decreased from 53% to 13% (P =.001). The cardiac index increased from mean 2.6 L/min per square meter (95% CI: 2.48-2. 72 L/min per square meter) to 3.1 L/min per square meter (95% CI: 2. 94-3.26 L/min per square meter; P =.001). The percentage of the patients with mild-to-moderate diastolic dysfunction decreased from 43% to 18% (P =.001). The left ventricular mass index was reduced by 42.4 g (95% CI: 35-50 g; P =.001). In comparison with biologic valves of the same size, mechanical valves produced a more pronounced reduction in mass index (overall difference 21.7 g; 95% CI: 37.1-6.4 g; P =.007) and a lower mean Doppler gradient (overall difference 4 mm Hg; 95% CI: 2-6 mm Hg; P =.0002). CONCLUSIONS: Patients undergoing aortic valve replacement had an improvement in functional status, as well as systolic and diastolic left ventricular function, and a reduction in left ventricular mass index, irrespective of prosthesis size and type. Mechanical valves are somewhat less obstructive than stented bioprosthetic valves of the same size. They are also associated with a concomitantly more pronounced reduction of left ventricular mass. PMID- 10384186 TI - Patent side branches do not affect coronary blood flow in internal thoracic artery-left anterior descending anastomosis: an experimental study. AB - BACKGROUND: It has been reported that large side branches of internal thoracic artery grafts may steal flow from the coronary circulation. Material an. METHODS: To assess the importance of the side branches, we measured the proximal and distal flow and pressures (mean subclavian artery pressure and mean arterial anastomotic pressure) at baseline and during infusion of adenosine (0.5 mg/kg/min) in 10 Landrace pigs in which an internal thoracic artery-left anterior descending anastomosis was constructed without interruption of the side branches. The difference between proximal and distal flow was considered to represent the blood flow of the internal thoracic artery side branches. Measurements were then repeated after surgical occlusion of all the side branches. RESULTS: At baseline, blood flow of the side branches represented 18% of the total flow in the proximal internal thoracic artery, and this percentage remained constant under the infusion of adenosine, which caused a 220% increase of the cardiac index and a 368% increase of the proximal flow. The infusion reduced the gradient along the left internal thoracic artery (mean subclavian artery pressure-mean arterial anastomotic pressure) from 15 to 10 mm Hg (P =.02) as the result of a lower mean subclavian artery pressure, although the mean arterial anastomotic pressure remained constant. Interruption of all the side branches resulted in a small and not significant increase in distal flow even after adenosine infusion. CONCLUSION: These observations suggest that blood flow in the side branches is minimal either at baseline and under combined systemic and coronary vasodilation. Clinically significant flow steal from the coronary circulation to the internal thoracic artery side branches seems then unlikely. PMID- 10384187 TI - Myocardial substrate uptake and oxidation during and after routine cardiac surgery. AB - OBJECTIVE: This study was designed to clarify whether myocardial substrate uptake and oxidation change after a period of hypothermic cardioplegic arrest during coronary artery bypass grafting procedures. METHODS: In 30 patients arterial and coronary sinus blood was sampled and coronary sinus flow measurements were performed before and after sternotomy and 10 minutes, 20 minutes, 50 minutes, and 6 hours after release of the aortic crossclamp. Measurement of free fatty acids, lactate, glucose, oxygen content, and carbon dioxide content in arterial and coronary sinus blood allowed calculations of myocardial substrate use, respiratory quotients, and myocardial oxidation rates of carbohydrates and fat. RESULTS: Uptake of free fatty acids and lactate was significant throughout the study and did not change in association with release of the crossclamp. Free fatty acid and lactate uptake measured 6 +/- 4 micromol/min and 23 +/- 26 micromol/min, respectively, before crossclamping compared with 8 +/- 7 micromol/min and 19 +/- 21 micromol/min, respectively, after release of the clamp. Glucose uptake was significant only during the first hour after crossclamp release and increased from 7 +/- 50 to 28 +/- 34 micromol/L after crossclamp release. Myocardial oxygen consumption did not change significantly (0.5 +/- 0.2 mmol/L compared with 0.35 +/- 0.2 mmol/L) after release of the crossclamp. Myocardial oxygen extraction ratio decreased from 58% +/- 8% to 41% +/- 13% after crossclamp release. Respiratory quotient increased after crossclamp release (0.85 +/- 0. 2 compared with 1.00 +/- 0.2), which implies that carbohydrate oxidation increased at the expense of free fatty acid oxidation. CONCLUSION: We conclude that hypothermic cardioplegic arrest during coronary artery bypass graft operations is associated with a transiently increased uptake and oxidation of carbohydrates during the immediate reperfusion phase. PMID- 10384189 TI - Pectus excavatum: increase of right ventricular systolic, diastolic, and stroke volumes after surgical repair. AB - OBJECTIVE: The study was undertaken to assess how a surgical correction of funnel chest modifies right ventricular structure and function. METHODS: Before and 6 months after surgery in 42 patients (27 male and 15 female patients, aged 5-31 years), a pectus index was calculated and echocardiographic examinations of the right ventricle were performed, with calculation of systolic, diastolic, and stroke volume indices. Right ventricular volume was estimated by subtracting the left ventricular volume from that of the entire heart. The values of the right ventricular volumes and the pectus index before and after the operation, as well as the changes in the indices, were compared. RESULTS: Statistically significant changes in the pectus index and the right ventricular volume indices after surgery were noted. No correlation was observed between the changes in the pectus index and the changes in any right ventricular volume indices. CONCLUSION: Surgical treatment of funnel chest causes an increase in right ventricular systolic, diastolic, and stroke volumes, although there is no correlation between these changes and the degree of sternocostal elevation. PMID- 10384188 TI - Left heart hypoplasia and neonatal aortic arch obstruction: is the Rhodes left ventricular adequacy score applicable? AB - OBJECTIVE: Although the influence of small left heart structures on outcome of a biventricular repair in neonatal critical aortic stenosis is well documented, little is known about its effect in neonates with aortic arch obstruction and coarctation. The purpose of this study was to evaluate the influence of small left heart structures on early and late results of repair and the ability to achieve a biventricular repair in neonates with coarctation and aortic arch obstruction. PATIENTS: Neonates included in this study had a left ventricular adequacy score (as proposed by Rhodes and associates for critical aortic stenosis) that would have predicted a need for a univentricular (Norwood) repair. All were ductus dependent but had antegrade ascending aortic flow and a small but nonstenotic aortic valve (<30 mm Hg gradient). Twenty neonates aged 10 +/- 9 days were identified for the study with weights averaging 3. 1 +/- 0.6 kg. Selected left heart measurements obtained by preoperative echocardiography included the following: aortic anulus 5.3 +/- 0.3 mm, mitral anulus 8.4 +/- 1.0 mm, transverse aortic arch 3.4 +/- 0.6 mm, and left ventricular volume 25 +/- 4 mL/m2. All patients underwent coarctation repair by resection and extended end-to-end anastomosis to enlarge the transverse arch as needed. Three patients underwent simultaneous pulmonary artery banding because of a hemodynamically significant ventricular septal defect. These 3 patients have subsequently had their defects successfully closed without mortality. RESULTS: There were no early or late deaths at a follow-up of 38 +/- 16 months after the operation. Three patients (3/20, 15%) have had to undergo reintervention with balloon aortoplasty because of recurrent coarctation (gradient > 20 mm Hg) in 2 and resection of subaortic stenosis in 1. Late follow-up in the remaining patients reveals 1 with moderate subaortic stenosis (gradient = 43 mm Hg), 2 with mild aortic stenosis (gradient < 30 mm Hg), and 2 with mild to moderate mitral stenosis. At late follow-up, 16 patients (16/20, 80%) are completely free of symptoms and 4 (4/20, 20%) have mild residual symptoms. CONCLUSIONS: Biventricular physiology can be successfully achieved in neonates with small left heart structures and aortic arch obstruction with minimal mortality and excellent late functional results. Standard echocardiographic measurements used to predict the need for a univentricular repair in critical aortic stenosis are not valid for the neonate with aortic arch obstruction. PMID- 10384190 TI - Commentary PMID- 10384191 TI - Clinical results of mitral valve repair by reconstructing artificial chordae tendineae in children. AB - OBJECTIVE: There are an increasing number of reports concerning mitral valve repair by reconstructing the chordae tendineae with the use of expanded polytetrafluoroethylene sutures in adults. However, little information is available about application or results of this technique in children. METHODS: Between January 1995 and December 1997, 16 children who had from moderate to severe mitral regurgitation mainly as the result of a prolapse of the anterior leaflet (age range, 5 months-12.8 years) underwent mitral valve repair by reconstruction of artificial chordae. Either unilateral or bilateral Kay-Reed annuloplasty was also performed to correct annular dilatation in all patients. RESULTS: No operative death or morbidity was observed. Before discharge, immediate postoperative echocardiography showed less than trivial mitral regurgitation in all patients. The follow-up was complete in all cases by a clinical examination and serial echocardiograms, and the median follow-up period was 14.8 months (range, 1.3-26.4 months). There were no valve-related events during the entire follow-up period. The degree of mitral regurgitation, estimated by echocardiography performed at recent follow-up period, was none in 5 patients, trivial in 10 patients, and mild in 1 patient. The diastolic and systolic dimensions of the left ventricle decreased and were 95.0% and 96.2% of the normal values, respectively. CONCLUSIONS: Although further investigations and long-term results are still called for, mitral valve repair by reconstruction of the artificial chordae was found to be safe and effective even in infants and children. PMID- 10384192 TI - Surgery for mitral valve disease in the pediatric age group. AB - OBJECTIVES: We reviewed a 20-year experience with the surgical treatment of mitral valve disease in the pediatric age group at our institution with 2 objectives: to clarify the long-term results over the last 2 decades and to evaluate the recent advances in mitral valve operation in children. METHODS: Since December 1978, 56 patients have undergone a total of 36 mitral valve repairs and 30 mitral valve replacements. Associated cardiac anomalies were present in 46 patients (82%), and concurrent repair of associated lesions was performed in 37 patients (66%). The age of the patients ranged from 3 months to 15 years (mean, 3.6 years) at mitral valve repair, and ranged from 2 months to 16 years (mean, 5.7 years) at mitral valve replacement. Mean follow-up period was 92.0 months (range, 1-235 months). RESULTS: There were 2 hospital deaths and 2 late deaths in patients who underwent mitral valve repair. Reoperation was performed in 4 patients. Three of these patients underwent mitral valve replacement because of residual mitral incompetence. No hospital deaths occurred in patients who underwent mitral valve replacement. Two late deaths occurred after mitral valve replacement. Six patients had a total of 10 episodes of prosthetic valve thrombosis. Thrombolytic therapy with urokinase was successful in all episodes without serious complications. Five patients required reoperations 49 to 141 months (mean, 78.4 months) after the initial valve replacement for relative prosthetic valve obstruction as the result of somatic growth. A valve 2 or 3 sizes larger than the original prostheses was inserted without death. Actuarial survival and freedom from cardiac events at 10 years after the operation were 87.2% and 72.7% in children who underwent mitral valve repair, and 90.3% and 67.3% for those children who underwent mitral valve replacement. CONCLUSIONS: The current risk of mitral valve operation in the pediatric age group is low, and the long-term results are satisfactory, irrespective of severe deformation of the mitral valve apparatus and associated complex cardiac anomalies. PMID- 10384194 TI - Bicaval and standard techniques in orthotopic heart transplantation: medium-term experience in cardiac performance and survival. AB - OBJECTIVE: The aim of this study was to compare the medium-term results of right heart pressures, tricuspid valve dysfunction, overall cardiac performance, and survival between the bicaval and standard techniques. METHOD: Between 1991 and 1997, 201 heart transplantations were performed in our center. Right heart catheterization was performed up to 12 months after transplantation. Echocardiography was used to assess left ventricular and tricuspid valve function. RESULT: The standard technique was used in 105 cases, and the bicaval technique was used in 96 cases. There was no difference in the age, preoperative parameters, pulmonary hemodynamics, or ischemic time between the 2 groups. Right atrial pressure (4.3 +/- 4.0 mm Hg for the bicaval vs 10.9 +/- 4.8 mm Hg for standard technique) and mean pulmonary artery pressure (17.5 +/- 5.3 mm Hg and 22.5 +/- 5.2 mm Hg, respectively) were lower for the bicaval recipients up to 12 months after the operation (P =.001 and. 01, respectively). Left ventricular ejection fraction was higher for the recipients of the bicaval technique up to the most recent measurement (P =.005). The prevalence of moderate or severe tricuspid regurgitation was higher in the recipients of the standard technique up to the most recent measurement (28% vs 7%; P =.02). The actuarial survival at 1, 3, and 5 years was 74%, 70%, and 62% for the recipients of the standard technique versus 87%, 82%, and 81% for the recipients of the bicaval technique (P <.03, <.04, and <.02, respectively). CONCLUSION: The bicaval technique maintains good left ventricular function, lower incidence and severity of tricuspid valve dysfunction, and improved survival compared with the standard technique. PMID- 10384193 TI - Preimplantation retrograde pneumoplegia in clinical lung transplantation. AB - OBJECTIVE: Retrograde pneumoplegia seems to improve early graft function in experimental and clinical lung transplantation. We evaluated the role of retrograde flushing in addition to antegrade pneumoplegia in clinical lung transplantation. METHODS: Fourteen patients undergoing lung transplantation were randomized into 2 groups: in group I we performed antegrade pulmonary artery flushing with alprostadil (prostaglandin E1) and modified Euro-Collins solution at the time of retrieval. In group II additional retrograde flushing through the pulmonary veins was performed at the back table, before reimplantation. Hemodynamic variables, mean airway pressure, and blood gas analysis were monitored at different time points. Postoperative volumetric monitoring was performed to assess extravascular lung water. The reimplantation response was assessed by a radiographic score; extubation time and intensive care unit stay were recorded. RESULTS: During retrograde flushing, blood and clots coming out from the pulmonary artery were observed; 2 lungs harvested from a donor with multiple bone fractures had fat emboli in the retrograde perfusate. Hemodynamic monitoring did not demonstrate any difference between the 2 groups. The ratio of arterial oxygen tension to inspired oxygen fraction, extravascular lung water, duration of intubation, and length of stay in the intensive care unit were improved in group II, but the differences did not reach statistical significance. Intrapulmonary shunt fraction was significantly improved in group II at each time point ( P =.02), as well as indexed alveolar-arterial oxygen tension gradient (P =.04), mean airway pressure (P =.04), and chest x-ray score ( P =.03). CONCLUSIONS: Preimplantation retrograde flushing is not detrimental and helps to improve early graft function. PMID- 10384195 TI - Long-term myocardial preservation: effects of hyperkalemia, sodium channel, and Na/K/2Cl cotransport inhibition on extracellular potassium accumulation during hypothermic storage. AB - OBJECTIVES: We previously demonstrated improved myocardial preservation with polarized (tetrodotoxin-induced), compared with depolarized (hyperkalemia induced), arrest and hypothermic storage. This study was undertaken to determine whether polarized arrest reduced ionic imbalance during ischemic storage and whether this was influenced by Na+/K +/2Cl- cotransport inhibition. METHODS: We used the isolated crystalloid perfused working rat heart preparation (1) to measure extracellular K+ accumulation (using a K+-sensitive intramyocardial electrode) during ischemic (control), depolarized (K+ 16 mmol/L), and polarized (tetrodotoxin, 22 micromol/L) arrest and hypothermic (7.5 degrees C) storage (5 hours), (2) to determine dose-dependent (0.1, 1.0, 10 and 100 micromol/L) effects of the Na +/K+/2Cl- cotransport inhibitor, furosemide, on extracellular K+ accumulation during polarized arrest and 7.5 degrees C storage, and (3) to correlate extracellular K+ accumulation to postischemic recovery of cardiac function. RESULTS: Characteristic triphasic profiles of extracellular K+ accumulation were observed in control and depolarized arrested hearts; a significantly attenuated profile with polarized arrested hearts demonstrated reduced extracellular K+ accumulation, correlating with higher postischemic function (recovery of aortic flow was 54% +/-4% [P =.01] compared with 39% +/-3% and 32% +/-3% in depolarized and control hearts, respectively). Furosemide (0.1, 1.0, 10, and 100 micromol/L) modified extracellular K+ accumulation by -18%, 38%, -0.2%, and +9%, respectively, after 30 minutes and by -4%, -27%, +31%, and +42%, respectively, after 5 hours of polarized storage. Recovery of aortic flow was 53% +/-4% (polarized arrest alone), 56% +/-8%, 70% +/-2% (P =.04 vs control), 69% +/-4% (P =.04 vs control), and 65% +/-3% ( P =. 04 vs control), respectively. CONCLUSIONS: Polarized arrest was associated with a reduced ionic imbalance (demonstrated by reduced extracellular K+ accumulation) and improved recovery of cardiac function. Further attenuation of extracellular K + accumulation (by furosemide) resulted in additional recovery. PMID- 10384196 TI - Superior protection in orthotopic rat lung transplantation with cyclic adenosine monophosphate and nitroglycerin-containing preservation solution. AB - BACKGROUND: Primary lung graft failure is common, and current lung preservation strategies are suboptimal. Because the decline in lung levels of cyclic adenosine monophosphate and cyclic guanosine monophosphate during preservation could enhance adhesiveness of endothelial cells for leukocytes as well as increase vascular permeability and vasoconstriction, we hypothesized that buttressing these levels by means of a preservation solution would significantly improve lung preservation. METHODS: An orthotopic rat left lung transplantation model was used. Lungs were harvested from male Lewis rats and preserved for 6 hours at 4 degrees C with (1) Euro-Collins solution (n = 8); (2) University of Wisconsin solution (n = 8); (3) low-potassium dextran glucose solution (n = 8); (4) Columbia University solution (n = 8), which contains a cyclic adenosine monophosphate analog (dibutyryl cyclic adenosine monophosphate) and a nitric oxide donor (nitroglycerin) to buttress cyclic guanosine monophosphate levels; or (5) Columbia University solution without cyclic adenosine monophosphate or nitroglycerin (n = 8). PaO2, pulmonary vascular resistance, and recipient survival were evaluated 30 minutes after left lung transplantation and removal of the nontransplanted right lung from the pulmonary circulation. RESULTS: Among all groups studied, grafts stored with Columbia University solution demonstrated the highest Pa O2 (355 +/- 25 mm Hg for Columbia University solution versus 95 +/- 22 mm Hg for Euro-Collins solution, P <.01, 172 +/- 55 mm Hg for University of Wisconsin solution, P <.05, 76 +/- 15 mm Hg for low-potassium dextran glucose solution, P <.01, and 82 +/- 25 mm Hg for Columbia University solution without cyclic adenosine monophosphate or nitroglycerin, P <.01) and the lowest pulmonary vascular resistances (1 +/- 0.2 mm Hg * mL-1 * min-1 for Columbia University solution versus 12 +/- 4 mm Hg * mL-1 * min-1 for Euro-Collins solution, P <.01, 9 +/- 2 mm Hg * mL-1 * min-1 for University of Wisconsin solution, 14 +/- 6 mm Hg * mL-1 * min-1 for low-potassium dextran glucose solution, P <.01, and 8 +/- 2 mm Hg * mL-1 * min-1 for Columbia University solution without cyclic adenosine monophosphate and nitroglycerin). These functional and hemodynamic improvements provided by Columbia University solution were accompanied by decreased graft leukostasis and decreased recipient tumor necrosis factor alpha and interleukin 1alpha levels compared with the other groups. In toto, these improvements translated into superior survival among recipients of Columbia University solution-preserved grafts (100% for Columbia University solution, 37% for Euro Collins solution, P <.01, 50% for University of Wisconsin solution, P <.05, 50% for low-potassium dextran glucose solution, P <.05, and 13% for Columbia University solution without cyclic adenosine monophosphate and nitroglycerin, P <.01). CONCLUSION: Nitroglycerin and cyclic adenosine monophosphate confer beneficial vascular effects that make Columbia University solution a superior lung preservation solution in a stringent rat lung transplantation model. PMID- 10384197 TI - The prognosis of surgically resected N2 non-small cell lung cancer: the importance of clinical N status. AB - BACKGROUND: Clinical trials dealing with multimodal strategy for N2 non-small cell lung cancer are now being watched with keen interest, and the feasibility of this strategy is to be confirmed. N2 lung cancer, however, is composed of several subgroups with different prognoses. The prognostic factors still remain controversial. METHODS: Between January 1986 and July 1997, 222 patients with lung cancer underwent surgical intervention at our institute; these patients were eventually given a diagnosis of metastasis to ipsilateral mediastinal lymph nodes. All patients underwent mediastinal lymph node dissection or sampling. Sixteen clinicopathologic factors were investigated by univariable and multivariable analyses to identify significant prognostic factors among resected N2 disease. Clinical N status was evaluated by computed tomographic scan. RESULTS: The overall 5-year survival was 27%. Multivariable analyses among overall patients revealed 4 significant prognostic factors (P <.05): clinical N2 status, incomplete resection, larger tumor size, and multiple diseased N2 nodes. Based on the result, 32 patients with both clinical N2 status and pathologic multiple N2 nodes showed a 5-year survival of 5%, whereas 76 patients with neither of the factors showed a 5-year survival of 57% (P <.001). CONCLUSION: The prognosis of surgically resected N2 disease varies tremendously according to the 4 significant prognostic factors. These factors should be clearly described in reporting clinical trials on N2 lung cancer. Clinical N status evaluated by computed tomographic scan should be 1 criterion to perform a clinical trial for N2 disease among a homogeneous population with respect to prognosis. PMID- 10384198 TI - Inhibition of the transcriptional activator protein nuclear factor kappaB prevents hemodynamic instability associated with the whole-body inflammatory response syndrome. AB - BACKGROUND: The transcription factor nuclear factor kappaB mediates the expression of a number of inflammatory genes involved in the whole-body inflammatory response to injury. We and others have found that dithiocarbamates specifically inhibit nuclear factor kappaB-mediated transcriptional activation in vitro. OBJECTIVE: We hypothesized that inhibition of nuclear factor kappaB with dithiocarbamate treatment in vivo would attenuate interleukin 1 alpha-mediated hypotension in a rabbit model of systemic inflammation. METHODS: New Zealand White rabbits were anesthetized and cannulated for continuous hemodynamic monitoring during 240 minutes. Rabbits were treated intravenously with either phosphate-buffered saline solution or 15 mg/kg of a dithiocarbamate, either pyrrolidine dithiocarbamate or proline dithiocarbamate, 60 minutes before the intravenous infusion of 5 micrograms/kg interleukin 1 alpha. Nuclear factor kappaB activation was evaluated by electrophoretic gel mobility shift assay of whole-tissue homogenates. RESULTS: Infusion of interleukin 1 alpha resulted in significant decreases in mean arterial pressure and systemic vascular resistance, both of which were prevented by treatment with dithiocarbamate. Pyrrolidine dithiocarbamate induced a significant metabolic acidosis, whereas proline dithiocarbamate did not. Nuclear factor kappaB-binding activity was increased within heart, lung, and liver tissue 4 hours after interleukin 1 alpha infusion. Treatment with dithiocarbamate resulted in decreased nuclear factor kappaB activation in lung and liver tissue with respect to that in control animals. CONCLUSIONS: These results demonstrate that nuclear factor kappaB is systemically activated during whole-body inflammation and that inhibition of nuclear factor kappaB in vivo attenuates interleukin 1 alpha-induced hypotension. Nuclear factor kappaB thus represents a potential therapeutic target in the treatment of hemodynamic instability associated with the whole-body inflammatory response. PMID- 10384199 TI - Intracellular free calcium accumulation in ferret vascular smooth muscle during crystalloid and blood cardioplegic infusions. AB - OBJECTIVE: The effects of magnesium- and potassium-based crystalloid and blood containing cardioplegic solutions on coronary smooth muscle intracellular free calcium ([Ca2+]i) accumulation and microvascular contractile function were examined. METHODS: Isolated ferret hearts were subjected to hyperkalemic (25 mmol/L K+) blood cardioplegic infusion, hypermagnesemic (25 mmol/L Mg2+, K+-free) crystalloid cardioplegic infusion, or hyperkalemic crystalloid cardioplegic infusion for 1 hour. Coronary arterioles were isolated, cannulated, and loaded with fura 2. Reactivity and [Ca2+]i were assessed with videomicroscopy. [Ca2+]i was measured at baseline and after application of 50 mmol/L KCl. In addition, [Ca2+]i and vascular contraction were measured during exposure to Mg2+ and K+ cardioplegic solution at both 4 degrees C and 37 degrees C. RESULTS: From a baseline [Ca2+]i of 177 +/- 52 nmol/L, K+ cardioplegic infusion (302 +/- 80 nmol/L potassium) markedly increased [Ca2+]i, whereas blood cardioplegic infusion (214 +/- 53 nmol/L) and Mg2+ cardioplegic infusion (180 +/- 42 nmol/L) did not alter [Ca2+]i. Although a difference between groups in percentage contraction after application of 50 mmol/L KCl was not observed, [Ca2+]i increased significantly more in vessels in the control group (764 +/- 327 nmol/L) and the K+ crystalloid cardioplegic infusion group (698 +/- 215 nmol/L) than in vessels in the blood cardioplegic infusion group (402 +/- 45 nmol/L) and the Mg2+ cardioplegic infusion group (389 +/- 80 nmol/L). Mg2+ cardioplegic solution induced no microvascular contraction at either 4 degrees C or 37 degrees C, nor was an increase in [Ca2+]i observed. K+ cardioplegic solution induced microvascular contraction at 37 degrees C but not at 4 degrees C; it increased [Ca2+]i at both 4 degrees C and 37 degrees C. CONCLUSION: An Mg2+-based cardioplegic solution, or appropriate Mg2+ or blood supplementation of a K+ crystalloid cardioplegic solution, may decrease the accumulation of [Ca2+]i in the vascular smooth muscle during ischemic arrest. PMID- 10384200 TI - Altered endothelium-derived hyperpolarizing factor-mediated endothelial function in coronary microarteries by St Thomas' Hospital solution. AB - OBJECTIVES: We examined the effect of St Thomas' Hospital solution on endothelium derived hyperpolarizing factor-mediated function in the porcine coronary microarteries with emphasis on the effect of temperature and washout time. METHODS: Microartery rings (diameter, 200-450 micrometers) were studied in myograph. The arteries were incubated in St Thomas' Hospital or Krebs solution (control) at 4 degrees C for 4 hours followed by 45 minutes (group Ia) or 90 minutes washout (group Ib) or at 22 degrees C for 1 hour followed by 45 minutes (group IIa) or 90 minutes washout (group IIb) and precontracted with -8.5 log M U 46619. The endothelium-derived hyperpolarizing factor-mediated relaxation to bradykinin was studied when endothelium-derived nitric oxide and prostaglandin I2 were inhibited with the presence of 7 micromol/L indomethacin and 300 micromol/L NG-nitro-L -arginine. RESULTS: After exposure to St Thomas' Hospital solution, the maximal endothelium-derived hyperpolarizing factor-mediated relaxation (percentage of the precontraction) was significantly reduced at either temperature after washout for 45 minutes (group Ia, 42.7% +/- 3.5% vs 69.0% +/- 5.3%; n = 9; P =.000; and group IIa, 12.3% +/- 1.6% vs 56.1% +/- 4. 4%; n = 8; P =.000) but fully recovered after washout for 90 minutes. The U46619-induced contraction force was also significantly reduced after washout for 45 minutes (P <.001) but fully recovered at 90 minutes. CONCLUSIONS: Under profound and moderate hypothermia, St Thomas' Hospital solution impairs endothelium-derived hyperpolarizing factor-mediated relaxation and smooth muscle contraction in the coronary microarteries. These effects exist during the reperfusion period for at least 45 minutes after exposure to St Thomas' Hospital solution and may account for the possible myocardial dysfunction during reperfusion. PMID- 10384201 TI - Effects of postischemic left ventricular pressure-volume unloading on contractile recovery and myocardial blood flow in the regionally stunned canine heart. AB - OBJECTIVE: Myocardial stunning remains a clinical problem without definitive therapy. This study tested the hypothesis that mechanical therapy with a ventricular assist device would accelerate recovery of contractility in stunned myocardium by increasing the postischemic myocardial blood flow. METHODS: Regional stunning was induced in dogs (25 kg) by 15 minutes of coronary occlusion and 180 minutes of reperfusion. One group (ventricular assist device; n = 10) was reperfused in conjunction with left ventricular unloading with a centrifugal-pump ventricular assist device. A second group (control; n = 8) underwent unmodified reperfusion. Hemodynamic and regional function data were acquired in all dogs with the heart in the working state before and during ischemia and after 180 minutes of reperfusion. Regional myocardial blood flow was measured at these same intervals and after 30 minutes of reperfusion, at which time the left ventricle was mechanically unloaded in animals with a ventricular assist device. RESULTS: Regional stunning was observed in all animals, but cardiogenic shock developed in none of them. After 180 minutes of reperfusion, animals with a ventricular assist device had greater systolic shortening in the risk segment than did control animals (11.5% +/- 2.8% vs 1.1% +/- 1.3%; P <.05) and had no differences in either the slope or x-axis intercept of regional preload recruitable stroke work relations compared with preischemic values. Differences in contractile recovery did not correlate, however, with postischemic myocardial blood flow. Hyperperfusion mediated by the ventricular assist device was not observed in either stunned or remote segments. CONCLUSIONS: Mechanical left ventricular unloading attenuates regional myocardial stunning within 3 hours in normotensive dogs, independent of effects on myocardial blood flow. The mechanism underlying this effect remains undefined, but these data support expanded use of mechanical therapy for stunned myocardium in clinical settings. PMID- 10384202 TI - Treatment of severe cardiac contusion with a left ventricular assist device in a patient with multiple trauma. PMID- 10384203 TI - Recombinant hirudin for extended aortic surgery in patients with heparin-induced thrombocytopenia. PMID- 10384204 TI - Kinetics of interleukin-11 release after cardiopulmonary bypass. PMID- 10384205 TI - Reversal of refractory hypotension with single-dose methylene blue after coronary artery bypass surgery. PMID- 10384206 TI - Stent fracture in a Carpentier-Edwards supra-annular porcine bioprosthesis implanted in the mitral position. PMID- 10384208 TI - Joint statement on redundant (duplicate) publication by the editors of the undersigned cardiothoracic journals PMID- 10384207 TI - Association of esophageal achalasia and pulmonary actinomycosis infection simulating bronchial neoplasm. PMID- 10384209 TI - Microscopic fungi in dwellings and their health implications in humans. AB - The article reviews the quantitative and qualitative incidence of microscopic filamentous fungi in dwellings, methods for their detection, mycotoxins, glucans and volatile organic compounds produced by microscopic fungi in the indoor air of homes. Characteristics and properties of the most important species of fungi in dwellings (Alternaria spp., Aspergillus spp., Cladosporium spp., Fusarium spp., Penicillium spp., Stachybotrys spp., and Wallemia spp.) and the health problems of occupants of the "moldy homes are also discussed. PMID- 10384210 TI - Occupational hazards of dentistry. AB - Dental professionals are susceptible to a number of occupational hazards. Relying on relevant literature, the present paper discusses selected occupational hazards - occupational biohazards, stressful situations, and latex hypersensitivity, as well as factors leading to the musculoskeletal system diseases and diseases of the peripheral nervous system. PMID- 10384211 TI - Impaired respiratory muscle function in chemical plant workers producing chlorfenvinphos. AB - All employees of a chemical plant division producing chlorfenvinphos were studied, i.e. 35 males aged 25-57 years (mean 42.1); their employment period ranged from 1-15 years (mean 9.0). Chronic bronchitis was diagnosed in 13 workers (37.1%). Mean air chlorfenvinphos concentrations in the work environment estimated with gas-liquid chromatography were from 0.0008-0.0018 mg/m3 (maximum allowable concentration according to Polish standards is 0. 01 mg/m3). The activity of erythrocyte acetylcholinesterase was similar to that observed in people who were not exposed to chemicals, however, a slightly lowered activity of plasma cholinesterase in the studied population was evidently the result of mild liver impairment. Spirometric investigations performed in the studied workers revealed slight alterations manifested by increased intrathoracic gas volume (ITGV) (the value of the index was 138.6% of the mean value, 24 workers with an abnormally high index), as well as by decreased specific airway conductance (sGaw); its mean value in the studied group was 58.5% of the mean standard (11 people showed an abnormal index). Substantial functional changes were found in the respiratory muscles. Maximal inspiratory pressures (MIP = 97. 2 +/- 28.3 cm H2O) as well as maximal expiratory pressures (MEP = 113.9 +/- 44.2 cm H2O) in the studied group were significantly lower (p < 0.01) as compared to those observed in the control group (MIP = 120.7 +/- 31.7; MEP = 154.4 +/- 40.2 cm H2O) of 22 males having similar cigarette smoking habit, without occupational exposure to chemicals. It was also found that the people who had worked for more than 10 years under conditions of exposure to chlorfenvinphos showed significantly lower (p < 0.05) values of maximal inspiratory pressure (87.2 +/- 28.06 cm H2O, n = 17) compared to the workers whose period of employment was shorter than 10 years (106.6 +/- 26.8 cm H2O, n = 18). The two groups were comparable with regard to age and smoking habits. The values of maximal expiratory pressures were similar in both groups. No essential disturbances in neuro-muscular transmission were observed; only in 3 workers (8.5%) the electrostimulating myasthenic test showed some disturbances in neuro-muscular transmission. It seems that respiratory muscles impairment in humans exposed to chlorfenvinphos results from changes in the metabolism and structure of muscles, and partly from lung hyperinflation. PMID- 10384212 TI - Use of a field portable X-Ray fluorescence analyzer to determine the concentration of lead and other metals in soil samples. AB - Field portable methods are often needed in risk characterization, assessment and management to rapidly determine metal concentrations in environmental samples. Examples are for determining: "hot spots" of soil contamination, whether dust wipe lead levels meet housing occupancy standards, and worker respiratory protection levels. For over 30 years portable X-Ray Fluorescence (XRF) analyzers have been available for the in situ, non-destructive, measurement of lead in paint. Recent advances made possible their use for analysis of airborne dust filter samples, soil, and dust wipes. Research at the University of Cincinnati with the NITON 700 Series XRF instrument (40 millicurie Cadmium 109 source, L X Rays) demonstrated its proficiency on air sample filters (NIOSH Method No. 7702, "Lead by Field Portable XRF; limit of detection 6 microg per sample; working range 17-1,500 microg/m3 air). Research with lead dust wipe samples from housing has also shown promising results. This XRF instrument was used in 1997 in Poland on copper smelter area soil samples with the cooperation of the Wroclaw Medical Academy and the Foundation for the Children from the Copper Basin (Legnica). Geometric mean soil lead concentrations were 200 ppm with the portable XRF, 201 ppm with laboratory-based XRF (Kevex) and 190 ppm using atomic absorption (AA). Correlations of field portable XRF and AA results were excellent for samples sieved to less than 125 micrometers with R-squared values of 0.997, 0.957, and 0.976 for lead, copper and zinc respectively. Similarly, correlations were excellent for soil sieved to less than 250 micrometers, where R-squared values were 0. 924, 0.973, and 0.937 for lead, copper and zinc, respectively. The field portable XRF instrument appears to be useful for the determination of soil pollution by these metals in industrial regions. PMID- 10384213 TI - Importance of sampling, extraction and preservation for the quantitation of biologically active endoto. AB - The influence of filter media, extraction solution and preservation method on detection of biologically active endotoxin in the LAL assay was studied with air samples collected from wastewater treatment plants. The four most common types of filters were used as collection media. The extraction solutions compared were nonpyrogenic water, KH2PO4-triethylamine and Trizma buffers. The effect of preservation on endotoxin air samples was ascertained by storing both the filters without extraction, and samples extracted in the collection day for a few weeks at various temperatures. Samples collected on glass fibre filters showed the highest amounts of detectable endotoxin, while the concentrations of endotoxin were significantly lower when cellulose-mixed esters, polycarbonate or polyvinyl chloride membrane filters were used for air sampling. After collection, the best efficiency for glass fibre filters was attained by extraction with nonpyrogenic water within 8 hours after sampling and storage of the extracts at 4 degrees C until they were analysed. If the filters were stored without extraction, the reduction in endotoxin levels of the sample was about 30% after 1 week preservation and about 70% after 2 weeks. The study shows that the effect of the filter material and preservation practice was significant. These factors play critical roles in assessing exposure to bacterial endotoxins within wastewater aerosols. PMID- 10384215 TI - Application of the classic Limulus test and the quantitative kinetic chromogenic LAL method for evaluation of endotoxin concentration in indoor air. AB - The classic (gel-clot procedure) Limulus test (CLT) and the quantitative kinetic chromogenic LAL method (KQCL) used for the evaluation of bacterial endotoxin concentration in the indoor air of dwellings were compared. The scientific procedure included analyses of 40 air samples supplemented by the analysis of 20 sample duplicates (selected at random) which were taken during the fall season from 10 flats located in 3 towns of the Upper Silesian region (southern Poland). The particulate aerosol probes were sampled by Harvard impactor and Casella sampler. The same samples were analyzed in the Netherlands using the quantitative kinetic chromogenic LAL method, and in Poland using the classic Limulus test. Comparison of both methods revealed that the quantitative kinetic chromogenic LAL method was more precise, with better reproducibility (the coefficient of variation between analyses of the main probe and its duplicate was over two times smaller in the KQCL method than in the CLT method), fully automated in the phase of analysis and data reading, and faster and more effective than the classic Limulus test. Nevertheless, on the basis of the obtained results, the usefulness of the classic Limulus method for assessment of the degree of pollution of indoor air with bacterial endotoxin seems to be confirmed as in the majority of examined samples (21 out 40) the results obtained by both methods were of the same order of magnitude, and in the remaining 19 samples did exceed one order of magnitude. Thus, the data received by means of the classic Limulus test may be regarded as acceptable. PMID- 10384214 TI - Exposure to airborne microorganisms and volatile organic compounds in different types of waste handling. AB - Occupational exposure of workers to airborne microorganisms and volatile organic compounds (VOC) in different types of waste treatment situations was examined during summer time. Microorganisms were collected as stationary samples using a six-stage Andersen impactor, while for VOCs both personal and stationary sampling was conducted. The exposure at the waste handling facility was considerably greater than at landfill sites or in waste collection. The concentrations of viable fungi were maximally 10(5) cfu/m3, and the concentrations of both total culturable bacteria and Gram-negative bacteria exceeded the proposed occupational exposure limit values (OELV), being 10(4) and 10(3) cfu/m3, respectively. Exposure to VOCs in the waste handling facility was three times higher than at the landfill sites, being at highest 3000 microg/m3, considered to be the limit for discomfort. The use of personal protective equipment at work, thorough hand washing and changing clothes after the work shift are strongly recommended in the waste handling facility and the landfill sites. PMID- 10384216 TI - Antigenic relationship between four airborne palm pollen grains from Calcutta, India. AB - The pollen grains of Areca catechu, Borassus flabellifer, Cocos nucifera and Phoenix sylvestris, all belonging to the family Aracaceae (Palmae), are airborne and found to be potent in causing human respiratory allergy. The present study was undertaken to discover the antigenic relationship, if any, in the four relevant palm pollen grains. The study was conducted by using Borassus and Phoenix antisera raised in rabbit. These antisera were used in rabbit IgG specific ELISA-inhibition and rocket immunoelectro-phoresis (RIE) assays for all four palm pollen extracts. In ELISA-inhibition, a distinct inhibition was obtained with comparable amount of soluble pollen protein. The RIE precipitin bands also revealed the presence of common antigenic components in the palm pollen. After isolation and purification, such common antigens may be useful in allergen immunotherapy in asthmatics. PMID- 10384217 TI - Seroepidemiologic study on the occurrence of antibodies against Yersinia enterocolitica and Yersinia pseudotuberculosis in urban and rural population of the Lublin region (eastern Poland). AB - The aim of this study was to assess the seroprevalence of antibodies against Yersinia in the rural and urban population and to determine the frequency of particular serotypes of Yersinia enterocolitica and Yersinia pseudotuberculosis. 472 sera were examined, 257 of rural inhabitants and 215 of urban inhabitants. The survey was carried out by passive hemagglutination test with the antigens of Yersinia serotypes considered pathogenic for humans: Y. enterocolitica 03, 05, 06, 08, 09 and Y. pseudotuberculosis I and III. In the examined rural population positive reactions to Yersinia antigens were significantly more frequent than in the examined urban population (42% versus 20%, p<0.0001). The most frequent reactions were against Y. enterocolitica serotypes 05 and 08. PMID- 10384218 TI - Occupational dermatoses in farmers - a proposal for diagnostic procedure. AB - The article presents a proposal for diagnostic procedures in cases of suspected occupational dermatosis in farmers. The process of certifying a disease as occupational is difficult because of lack of the monitoring of occupational risks in private farms; moreover, there is no compulsory medical assessment before one starts work as a farmer. Many patients meet an occupational health professional for the first time when the disease is already advanced and legal action towards obtaining an occupational rent has already been issued. In these circumstances, confirming or rejecting the possible occupational etiology of a given dermatosis is very difficult. This article presents a diagnostic procedure which has been devised by the author and used with some success for two years at the Institute of Agricultural Medicine, Lublin, Poland. PMID- 10384219 TI - Comparative analysis of pollen fall at three sites in south-eastern Poland. AB - The aim of the research was to study spatial variations in the abundance and seasonal patterns of pollen fall. The investigation was carried out at three sites of different land use during two pollen seasons (1995 and 1996). The sites were located in an average town (Ostrowiec Sw.), in a village (Brzostowa) and in the open area near Ozarow. With the use of the gravimetric method, 55 taxa of sporomorphs were determined. There were small differences in seasonal incidence and in total pollen sums between the three sites. The differences in the abundance and percentage values of chosen pollen taxa (Populus, Fraxinus, Pinus, Poaceae) were bigger. Tree and shrub pollen dominated in pollen fall at the three sites. The preliminary results suggest that the pollen data differed more between the years than between the sites, which was probably due to meteorological patterns. PMID- 10384220 TI - Injury analysis and prevention in the developing countries. PMID- 10384221 TI - A sociological case study of occupational accidents in the Brazilian petrochemical industry. AB - This is a case study of accidents occurring in a petrochemical plant located in South Eastern Brazil. It was based on comprehensive interviews with engineers and workers concerning major accidents. Information collected during the interviews highlighted the contrasts in how management and workers interpret industrial accidents. Some implications for actions were discussed. PMID- 10384222 TI - A survey on management perspectives of the state of workplace health and safety practices in Kenya. AB - A baseline survey was conducted in 1995 on management perspectives of occupational health and safety (OHS) structures and practices in Kenya. This was achieved by interviewing management and supervisory staff attending 1 week multi disciplinary courses that were organized by the Federation of Kenya Employers (FKE) and the International Labour Office (ILO) at hotel venues in Kenya. The purpose of the survey was to gain some insight into work safety conditions in Kenya and to assess the potential for a new OHS manual to meet existing knowledge gaps. The manual was locally developed in 1993/4 by Kenyan OHS experts in collaboration with colleagues from the Swedish National Institute for Working Life. Results of the survey from 65 participants indicated that most workplace managers were not familiar with the Kenyan work safety legislation. Work injuries were largely attributable to working with dangerous machinery. Occupational diseases and HIV/AIDS were cited as other causes of workplace morbidity and mortality. Although most respondents (70%) were satisfied with their work safety conditions, only 37% said their workplaces were annually audited by labour inspectors while 45% said injured workers were not treated well by management. Many workplaces (65%) violated the mandatory legal requirement on the establishment of health and safety committees. The OHS resource person and course content were rated highly by most respondents (96%). The foregoing results provided the basis of a needs analysis for future OHS programs in Kenya. PMID- 10384223 TI - Occupational injuries in Shunde City--a county undergoing rapid economic change in southern China. AB - A survey was carried out in Shunde City of Southern China to look into the occurrence of occupational injuries in the 'township industries' during the years 1989-1993. A total of 981 major injuries and 159 fatal injuries were recorded for 602,533 person years over the 5 years, giving a major injury rate of 1.63/1000 per year and a fatal injury rate of 0.26/1000 per year. Both the major injury rate and the fatal injury rate increased from 1989 to 1993. Males had a higher fatal injury rate while females had a higher major injury rate. The majority of the injuries occurred in the younger age group with the 20-29 age group taking up more than half of the total casualties. Industries having high injury rates included building and construction, furniture, paper mill and printing. Electrical appliances manufacturing, sewing and shoe making were the industries having low injury rates. Enterprises with foreign capitals and those managed by individuals also had lower injury rates. Machinery injuries were the most common type of major injuries while electrocution, being hit by heavy objects and falling from height accounted for over 60% of fatal injuries. The implications of the differential distributions of the occurrence of the injures were discussed. PMID- 10384224 TI - Urban development and traffic accidents in Brazil. AB - Brazil started to experience high traffic accident rates since the 1960s, when road transportation began to be dominant and the number of motorized vehicles increased sharply. The severity of the problem was also related to the fast and uncontrolled urban growth, which allowed for the organization of an inherently dangerous circulation space, characterized by a complex pattern of traffic conflicts. With respect to traffic accidents, the results have been highly negative: national statistics report about 28,000 fatalities a year, with more than 340,000 injured people. The paper analyses current patterns of traffic accidents in urban areas, using the data from two of the three largest cities in the country. Hospital data is analyzed to understand the true social impacts of such accidents. Several causal factors are identified, at the political, cultural and technical sides. Challenges to change current conditions are also identified. Practical measures are suggested, especially the change of the dangerous environment and the need to improve the technical approach to the problem, education programs and enforcement logistics. PMID- 10384225 TI - Risky behavior of bus commuters and bus drivers in Karachi, Pakistan. AB - Buses account for a disproportionate number of road traffic accident fatalities in Karachi, Pakistan and other developing countries. Potentially dangerous bus driving and commuting practices that increase risk of road accidents and the effect of traffic police on bus behavior are evaluated. A total of 250 episodes each of disembarking and embarking commuters, buses stopping and moving on the road at ten of Karachi's highest risk intersections for traffic injuries were observed. Of the disembarking passengers, 33% did not wait for the bus to stop; 54% stepped off in the center of the road and 84% did not look out for traffic. Among the embarking commuters, 38% got on moving buses; 73% climbed on buses filled to their outer foot boards and 83% waited for buses on the street. Males were more likely than females to jump off a moving bus (43% versus 1.6%, P < 0.001), get on a moving bus (49% versus 12%, P < 0.001), and run to catch a bus (45% versus 8%, P < 0.001). At the bus stops, 30% of the buses did not stop completely; 46% stopped away from the stop and 79% stopped in the center of the road. Where traffic police were present buses were more likely to race (9% versus 3%, P = 0.05) and to cut off other vehicles (13% versus 2%, P = 0.001) than where police were absent. Risky behavior is common among both Karachi bus drivers and bus commuters. The traditional efforts to regulate bus traffic through traffic police is ineffective. PMID- 10384226 TI - Pedestrian environment and behavior in Karachi, Pakistan. AB - Pedestrian road traffic accidents (RTAs) are responsible for a substantial number of injuries and deaths in Karachi. To better understand the situations facing pedestrians we selected ten of Karachi's highest risk locations for pedestrian RTAs and observed 250 pedestrians for each of three activities--crossing the street, walking on the street, and walking on the sidewalk. We also observed the extent and effect of street and sidewalk encroachments. A total of 35% of the pedestrians crossing the street caused traffic to swerve to avoid them. Pedestrians crossing one lane at a time were 2.9 times more likely to cause the traffic to swerve than pedestrians who crossed the whole street at once (53 vs. 18%, RR = 2.9, 95% CI = 1.9-4.3). Pedestrians crossing in a group were 1.8 times more likely to cause traffic to swerve compared to those crossing singly (49 vs. 28%, RR = 1.8, 95% CI = 1.3-2.5, P = 0.001). A total of 36% ran while crossing and were 1.8 times more likely to cause traffic to swerve than those who walked (48 vs. 27%, RR = 1.8, 95% CI = 1.3-2.5). An average of 77% of the sidewalk width was blocked by encroachments which forced pedestrians to step on the road resulting in vehicles swerving. An average 33% of the street width was blocked by illegally parked vehicles. Pedestrians in Karachi indulge in risky behaviors. Encroachments on streets and sidewalks compound the problem. Piloting efforts to modify pedestrian behavior and the environment they negotiate should be considered to reduce pedestrian deaths. PMID- 10384227 TI - An evaluation of sensitivity and specificity of blood alcohol concentrations obtained by a breathalyser survey in a casualty department in Kenya. AB - Whereas breathalysers have been shown to provide blood alcohol concentration (BAC) measurements comparable to those obtained by gas chromatography, such evidence has not been reported in low and middle income countries where measures for preventing alcohol-related injuries are virtually non-existent. Before promoting any method of blood alcohol evaluation, as a routine procedure for monitoring the association of alcohol with different types of injuries in Kenya, we sought to assess the reliability and validity of blood alcohol results obtained by a breathalyser, using gas chromatography analysis values as the reference, in a sample of 179 trauma-affected adults presenting to casualty departments. No differences in proportions of subjects with high levels of blood alcohol (equal to or greater than 50 mg%) were detected by breath and blood test procedures (58.7 vs 60.3%). Breathalyser readings yielded high levels of sensitivity and specificity (97.2 and 100%, respectively) with optimal positive and negative predictive values (100 and 95.9%, respectively) at higher BACs (> or = 50 mg%). The study thus reaffirms that breathalyser tests are of value in detecting high blood alcohol levels and can be used to rapidly identify intoxicated subjects. The procedure is easy to perform and can be used for monitoring the association between blood alcohol level and driving in low-income developing countries. PMID- 10384228 TI - Economic costs of traffic accidents in Jordan. AB - The objectives of this study were to estimate the economic costs of traffic accidents in Jordan during the year of 1996 and to derive unit accident costs for various accident severity levels. The related data were acquired from different sources, including traffic police records, insurance companies, private hospitals and medical centers. In this study, a framework for applying unit casualty class costs, unit property damage cost, as well as police activities and insurance administration costs to accidents of various severity levels was suggested. The loss-of-output, the loss quality of life, the community and family losses, the temporary and permanent losses, and hospitalization and medical treatment costs were estimated in computing the unit cost for fatalities or injuries of different casualty classes. The vehicle repair cost, detention period cost, and public and private costs were accounted for in estimating the unit cost of property damages. The results indicate that the 1996 traffic accidents cost the country about JD 103 million ($US 146.3 million). PMID- 10384229 TI - Epidemiology of transport-related injuries in Ghana. AB - To better elucidate the incidence, characteristics, and consequences of transport related injuries in a less developed country in Africa, we undertook an epidemiologic survey in Ghana. A total of 21,105 persons were surveyed, in both an urban area (Kumasi, n = 11,663) and a rural area (Brong-Ahafo, n = 9442). In the preceding year, a total of 656 injuries were reported in the urban area and 928 injuries reported in the rural area. Transport-related mechanisms accounted for 16% of all injuries in the urban and 10% of all injuries in the rural area. The annual incidence of transport-related injuries was almost identical in the two settings, 997/100,000 persons in the urban area and 941/100,000 in the rural area. In both settings, transport-related injuries were more severe than other types of injuries in terms of mortality, length of disability, and economic consequences. In the urban area, the most common transport-related mechanisms were either to passengers involved in crashes of mini-buses or taxis (29%) or to pedestrians struck by these vehicles (21%). In the rural area, the most common transport-related mechanisms were bicycle crashes. The second most common rural mechanisms were motor vehicle crashes, which were the most severe and which involved commercial (83%) rather than private vehicles. Prevention strategies need to be different from those in developed countries and should target commercial drivers more than private road users. PMID- 10384230 TI - Airbags: an exploratory survey of public knowledge and attitudes. AB - The present study examines public knowledge and opinion in the United States on issues related to airbag safety. Data were obtained through a national random digit-dial telephone survey of 1005 people living in the contiguous 48 United States. A majority of respondents (1) know that airbags can harm drivers seated too close to the steering wheel; (2) know that rear-facing infant seats should not be placed in the front seat of a car with passenger-side airbags; and (3) know that airbags are saving more lives of women drivers than are being lost. However, most respondents did not know that (1) airbags are killing more children than they are saving; (2) airbags can injure properly belted drivers; and (3) the majority of the lives saved by airbags have been among people who were not wearing safety belts. Knowledge of airbag risks to children and properly belted drivers was significantly associated with a less favorable attitude toward airbags, and with opposition toward the law mandating airbags on all new cars. Drivers of vehicles equipped with airbags held more favorable attitudes toward airbag technology. Further analysis suggests that as the public begins to understand the risks associated with airbags, the current high level of public support for the technology and the mandatory regulation may decline. PMID- 10384231 TI - Compatibility problems in frontal, side, single car collisions and car-to pedestrian accidents in Japan. AB - Compatibility problems in car-to-car frontal, side, single car and car-to pedestrian collisions in Japan are discussed using traffic accident data. The number of serious and fatal injuries is investigated for the subject car and other cars, which are categorized by their class and mass. The aggressivity of the cars is calculated by the number of fatalities, fatality rates and by the number of car registrations. The results show that in car-to-car frontal collisions, cars with a mass of 1150 kg are the most compatible among the current car population. In both car-to-car frontal and side collisions, the sports utility vehicle and mini car are found to be the most incompatible car types with high and low aggressivity, respectively. On the other hand, the accident data show that the wagon and midsize sedan are the most compatible car types. The compatibility of fixed objects in the road environment with cars and cars with pedestrians is also discussed. In a single car collision with a fixed object, the guardrail is the most compatible object and can reduce the fatality rate on prefecture roads by about 60%. The front geometry of the car has large effect on compatibility with a pedestrian. PMID- 10384232 TI - The influence of head restraint and occupant factors on peak head/neck kinematics in low-speed rear-end collisions. AB - Prior two-way analyses of variance showed that the peak kinematic response of the head and neck of subjects exposed to low-speed rear-end collisions was related to speed change and gender, however potential reasons for this gender dependence were not determined. Using multiple linear regression, this study further examined these response data to determine the relative influence of specific factors, including subject anthropometry, neck strength, cervical range of motion, seated posture and head restraint position, which may have been responsible for the previously-observed gender dependence. The results of this analysis showed that vehicle speed change and relative head restraint position explained the largest proportion of the observed variation in peak occupant kinematic response. Seated posture measures also explained some of the variation in kinematic response. The current analysis prioritizes which variables to explore more thoroughly in future research and which variables should be carefully controlled in future studies. PMID- 10384233 TI - The characteristics and use patterns of all-terrain vehicle drivers in the United States. AB - The consent decrees between the US Consumer Product Safety Commission and the major distributors of all-terrain vehicles (ATV), which were designed to address ATV-related injuries and deaths, expired in April, 1998. While national estimates of nonfatal and fatal injuries involving ATVs declined after the consent decrees went into effect 10 years ago, the injury estimates have stabilized in recent years. To gain a better understanding of current ATV use patterns, the CPSC sponsored a national probability survey of ATV drivers in the fall of 1997. The survey was designed to collect information about the characteristics and use patterns of ATV drivers and to quantify the numbers and types of ATVs in use. It employed a single stage list-assisted random-digit-dial sample design. This article describes the results of the survey, and discusses long term ATV usage trends. PMID- 10384234 TI - Picornaviruses: epitopes, canyons, and pockets. PMID- 10384235 TI - Naturally occurring variants of hepatitis B virus. PMID- 10384236 TI - Kaposi's sarcoma-associated herpesvirus: epidemiology, virology, and molecular biology. PMID- 10384237 TI - Role of chemokine receptors in HIV-1 infection and pathogenesis. PMID- 10384238 TI - Nuclear import of human immunodeficiency virus type-1 preintegration complexes. AB - Following infection-mediated entry into the cytoplasm, retroviral cores form large nucleoprotein complexes (PICs) which undergo reverse transcription and, ultimately, catalyze provirus formation. The ability of these complexes to be specifically imported into the nucleus via NPCs explains why nondividing cells can be productively infected with lentiviruses such as HIV-1, whereas productive infection by the oncoretrovirus MLV is restricted to proliferating cells. Current evidence suggests that virally encoded protein components of the HIV-1 PIC, in particular IN and Vpr, act in concert to target these complexes for nuclear import by recruiting cellular import factors and interacting with the NPCs. Here have we reviewed recent advances made in this complex and fascinating area of HIV 1 biology and have discussed them in relation to models for postentry nuclear import in other retroviral and nonretroviral systems. PMID- 10384239 TI - Host proteins in retroviral cDNA integration. PMID- 10384240 TI - HIV integrase structure and function. AB - HIV integrase consists of three domains, the structures of which have been individually determined by X-ray crystallography or NMR spectroscopy. The core domain, spanning residues 50-212, is responsible for the catalytic activity of the enzyme. The crystal structure of a dimer of this domain shows similarity to other proteins that carry out polynucleotidyl transfer, including MuA transposase and RNase H. The small N-terminal domain folds into a dimeric helix-turn-helix structure, which is stabilized by the coordination of zinc with conserved His and Cys residues. The function of this domain is unclear; however, it is required for integration activity and enhances tetramerization in the context of the full length integrase. The C-terminal domain, which has an SH3-like fold, is involved in DNA binding. The structure of this domain reveals a large saddle-shaped cleft that is formed by dimerization. This cleft contains a number of positively charged residues, and its dimensions are appropriate for accommodating a double stranded DNA helix. Although the C-terminal domain was originally believed to be involved in target DNA binding, more recent evidence suggests that it may bind to both the ends of the viral DNA and to the target DNA. Although the individual domain structures provide some insights into the function of the protein, a more detailed understanding of the complete mechanism by which integrase binds, cleaves, and transfers DNA requires a greater knowledge of how these domains are arranged in the active multimer. PMID- 10384241 TI - Crystal structures of catalytic core domains of retroviral integrases and role of divalent cations in enzymatic activity. AB - Crystal structures of the enzymatically competent catalytic domains of HIV-1 and ASV IN have been solved in the last few years. The structure of HIV-1 IN has been described only for apoenzyme and for a complex with Mg2+, whereas the structure of ASV IN has been presented as the apoenzyme, in the presence of divalent cations (Mn2+, Mg2+, Ca2+, Zn2+, and Cd2+), and with an inhibitor. A single ion of Mn2+, Mg2+, or Ca2+ interacts with the two aspartate side chains of the D,D(35)E catalytic center in octahedral coordination with four water molecules. However, two ions of Zn2+ or Cd2+ bind to the active site of IN with tetrahedral and octahedral coordination, respectively. Only small adjustments take place in the active site of ASV IN on binding of the metal cofactor(s), which are absolutely required for the activity of this enzyme. The placement of the side chains and metal ions in the active site is very similar to that observed even in distant members of this superfamily of polynucleotidyltransferases. Here the role of divalent cations in the enzymatic activity of IN and the search for inhibitors of this enzyme are discussed. PMID- 10384242 TI - HIV-1 integrase: structural organization, conformational changes, and catalysis. AB - Integrase comprises three domains capable of folding independently and whose three-dimensional structures are known. However, the manner in which the N terminal, catalytic core, and C-terminal domains interact in the holoenzyme remains obscure. Catalytically active recombinant IN can exist in a dynamic equilibrium of monomers, dimers, tetramers, and higher order species. Numerous studies indicate that the enzyme functions as a multimer, minimally a dimer. The IN proteins from HIV-1 and ASV have been studied most carefully with respect to the structural basis of catalysis. Although the active site of ASV IN does not undergo significant conformational changes on binding the required metal cofactor, that of HIV-1 IN does. The reversible, metal-induced conformational change in HIV-1 IN impairs the binding of some anti-HIV-1 IN monoclonal antibodies to the enzyme and results in differential susceptibility of the protein to proteolysis. This active site-mediated conformational change reorganizes the catalytic core and C-terminal domains and appears to promote an interaction that is favorable for catalysis. Other metal-dependent structural changes in HIV-1 IN include the promotion of interactions between the N terminal and the catalytic core domains and the induction of tetramers by zinc ions. The end result of these metal-induced changes is apparently the induction of an activated holoenzyme that can form a stable ternary integrase-metal-DNA complex. These structural changes, which appear to be crucial for optimum catalysis in HIV 1 IN, do not occur in ASV IN. The structural changes observed in HIV-1 IN may serve to recruit the catalytic machinery in this enzyme to a conformation that is native for ASV IN. PMID- 10384243 TI - Substrate recognition by retroviral integrases. AB - Substrate recognition by the retroviral IN enzyme is critical for retroviral integration. To catalyze this recombination event, IN must recognize and act on two types of substrates, viral DNA and host DNA, yet the necessary interactions exhibit markedly different degrees of specificity. Although particular sequences at the viral DNA termini are recognized by IN, many host DNA sequences can serve as the target for integration. Over the last decade, both in vitro and in vivo data have contributed to our understanding of how IN recognizes its substrates. This review provides an overview of the sequence and structure requirements for recognition of viral and host DNA by different retroviral INs and discusses recent progress in mapping protein domains involved in these interactions. PMID- 10384244 TI - Reconstitution of concerted DNA integration with purified components. PMID- 10384245 TI - In vivo analysis of retroviral integrase structure and function. AB - There are two retroviral integration loci. One encodes the transacting IN protein, which is cleaved from the carboxyl terminus of the Gag-Pol polyprotein precursor during virus assembly. The second locus is the cis-acting attachment (att) site, comprising the terminal sequences at the U3 and U5 ends of linear viral cDNA. Integrase and att site mutant viruses can be blocked at different steps of the viral replication cycle. Class I IN mutants are blocked specifically at the integration step. Class II IN mutants, on the other hand, display pleiotropic defects, most notably in virion morphogenesis and/or reverse transcription. Mutations in the U5 end att site can also disrupt reverse transcription in addition to integration. It is prudent to use caution when interpreting results of in vivo mutagenesis experiments that target retroviral IN and the att site. PMID- 10384246 TI - Inhibitors of human immunodeficiency virus integrase. AB - Integration of the viral DNA into a host cell chromosome is an essential step for HIV replication and maintenance of persistent infection. Two viral factors are essential for integration: the viral DNA termini (the att sites) and IN. Accruing knowledge of the IN structure, catalytic mechanisms, and interactions with other proteins can be used to design strategies to block integration. A large number of inhibitors have been identified that can be used as leads for the development of potent and selective anti-IN drugs with antiviral activity. PMID- 10384247 TI - Comparison of biofeedback and relaxation in the treatment of pediatric headache and the influence of parent involvement on outcome. AB - The relative efficacy of EMG-frontalis feedback and progressive relaxation was examined in children with tension-type or combined headaches (8-14 yrs. old). Furthermore, the influence of parent involvement, in the form of a three-session educational approach, on training outcome was systematically explored (2 x 2 factor design). Fifty children took part in the study, 40 were randomly assigned to the four different treatment conditions, 10 children participated in the self monitoring control group. The training comprised 6 sessions of 1 hr each in the relaxation treatment and 12 sessions of 1/2 hr duration in the biofeedback group. Headache diaries were kept by children and parents for 4-week period prior to therapy, and for a similar length of time at post-treatment and follow-up (6 months). Multivariate analyses of variance on the headache diary data yield no significant main or interaction effects of treatment format or of parent involvement, but only a main effect of period, indicating a general efficacy of the four treatment conditions. At follow-up the reduction of headache activity is even more prominent. A different evaluative approach points to the superiority of biofeedback revealing a mean effect size for biofeedback training that reflects a good to excellent improvement rate. Correlations between headache data from children and parents are high. PMID- 10384248 TI - Psychological changes accompanying and mediating stress-management training for essential hypertension. AB - In a previous controlled study, 21 participants with essential hypertension were treated with a program based on education, relaxation and D'Zurilla problem solving training, and another 21 participants were assigned to a waiting list control condition (Garcia-Vera, Labrador, & Sanz, 1997). In this report, the pre post-treatment psychological changes accompanying those conditions were examined with the Jenkins Activity Survey, the Rosenbaum Self-Control Schedule, the Spielberger State-Trait Anxiety Inventory, and the D'Zurilla-Nezu Social Problem Solving Inventory. Treatment yielded significant psychological changes that included an increase of problem-solving abilities. Moreover, correlation and multiple regression analyses revealed that, when clinic blood pressure (BP) values were considered, increases in problem-solving abilities were correlated with systolic and diastolic BP reductions for participants in the stress management condition, and they mediated partially the antihypertensive effects of stress-management training on BP. No significant correlations were found between psychological changes and self-measured systolic or diastolic BP reductions. PMID- 10384249 TI - Electroencephalographic and psychometric differences between boys with and without attention-deficit/Hyperactivity disorder (ADHD): a pilot study. AB - Attention-Deficit/Hyperactivity Disorder (ADHD) is reported to have an incidence of 3-5%, and is associated with a variety of interpersonal, academic, and social problem behaviors. There is controversy as to whether ADHD is a learned behavioral or brain dysfunction. Research has explored a variety of measures to assess behavioral and brain dysfunctions in this population, with no consistent and clearly diagnostic results. We investigated whether a new psychometric and a new electroencephalographic procedure would clearly differentiate ADHD. The psychometric was based on DSM-IV criteria and the EEG measure was based on the assumption that ADHD interferes with cognitive transition from one discrete task to another. Parents of four ADHD boys (ages 8-12) and four age- and interest matched non-ADHD boys completed the ADHD Symptom Inventory, while their sons' EEG was monitored during viewing of a video and reading of a book. For the ADHD boys, this was repeated a second time, 3 months later, to assess test-retest reliability. Both the psychometric and the EEG measures clearly differentiated the two samples (p's < .01) with no overlap in scores, were reliable over 3 months (r = .87), and were significantly correlated with one another (r = .85). While a small sample size, these robust, related and reliable findings suggest that both the psychometric and the psychophysiological EEG measures deserve further replication and exploration. PMID- 10384250 TI - Behavioral psychophysiological intervention in a mentally retarded epileptic patient with brain lesion. AB - Behavioral psychophysiological treatment entailing Slow Cortical Potential (SCP) biofeedback training and behavioral self-control training was conducted with a 27 year-old male epileptic patient (seizures for 23 years) with Wechsler IQ 64 who underwent callosotomy. The patient had 12/week secondary generalized tonic-clonic seizures. The treatment, consisting of 43 SCP training sessions and 22 behavioral control sessions, yielded a highly significant reduction of seizure frequency to about 7.5/week; such a decrease had never been observed after administration of new anticonvulsant drugs, nor after the callosotomy. During SCP feedback training, the patient was able to produce highly-significant cortical differentiation of SCPs of about 4 microV. In addition, he developed several new behaviors indicating growing ability of self-perception and self-regulation. These findings suggest that a combination of SCP biofeedback with behavioral treatment of epilepsy can be used even in mentally retarded patients with organic brain disorders. PMID- 10384251 TI - [Italy: a national health service with a tumultuous history]. AB - During the 70's, various political forces succeeded to combine their efforts and launched a health reform. The system based on sickness insurance funds each serving a specific social or occupational group was replaced by a universal system covering all the citizens. During the 80's, the reformist context disappeared, the basic units of the new system (U.S.L.) became extremely politicized, resources subsidized by the State became scarce, the people frustrated. At the beginning of the 90's, a new reform was launched, aimed at raising the management level of the USL (ambulatory care) and the large hospitals. At the same time, more financial and managerial responsibilities were transferred to the regions. During these decades, the Italian reform attempted to implement several foreign models. But a long time was wasted in debates and discussions. When the implementation started, these models became outdated and the authors inspiring the reform disappeared themselves from the stage. PMID- 10384252 TI - Hypertensive drug demand and reimbursement in public and private systems: a French-US comparison. PMID- 10384253 TI - [Vasectomy in Quebec: 1977-1997]. AB - Vasectomy is recognized as an efficient method of contraception. However, some recent findings have suggested that vasectomy may raise a potential health risk, notably related to prostate cancer. The goal of the present study is to evaluate the spatial and temporal evolution of this surgical practice in Quebec (Canada). Using the registry of the Regie de l'assurance-maladie du Quebec, we describe the course of vasectomy practice during the past 21 years. We also estimate the frequency of vasectomy procedures on the regional scale. Observed and expected rates of vasectomy and a synthetic index of vasectomy allow us to evaluate its spatial and temporal evolution. The results show a similar annual growth rate of this practice during the last 10 years. The mean age at the time of vasectomy of the study subjects (N = 332,000) is 35 years old. However recently, this mean age seems to be more representative because of the lower proportions of younger (< 25 years old) and older (> 50 years old) patients at the time of vasectomy. In addition, we observe some clear differences concerning the vasectomy rates between specific geographical sectors; the rates being significantly lower (p < 0.01) in urban and some rural areas. The highest rates are found in the suburbs of Montreal and Quebec City. There are also some local variations of the practice, which are difficult to interpret. In this article, we compare our results to those for the rest of Canada and the United States. We note that the rates are generally similar to those for the Province of Quebec. Besides, we discuss about the reasons for strong spatial variations of the surgery. PMID- 10384254 TI - [The Federation for International Cooperation of Health Services and Systems Research Centers: a decade of activity (1988-1998)]. PMID- 10384256 TI - Synaptic potentiation induced by a protein factor in cultured cerebral neurons. AB - 1. We reported in a previous paper that long-lasting enhancement of spontaneous excitatory post synaptic currents (SEPSCs) in cultured chick cerebral neurons was induced by exposure to a conditioned medium (CM) prepared by Mg(2+)-free treatment of neurons. This suggested that the CM contained a diffusible factor(s) for the potentiation. 2. In this paper, the factor(s) was shown to be a protein(s) by heat and trypsin treatment of the CM. 3. The factor induced the potentiation within 5 min, but it was not required for maintenance of increased SEPSCs. 4. The factors in CM induced the potentiation without protein synthesis. 5. Protein synthesis at least in postsynaptic neurons, was indispensable to induce the potentiation by the Mg(2+)-free condition. PMID- 10384255 TI - Angiotensin and cerebral blood flow. PMID- 10384257 TI - Nerve growth factor inducer, 4-methyl catechol, potentiates central sensitization associated with acceleration of spinal glutamate release after mustard oil paw injection in rats. AB - 1. In rats, injection of mustard oil (MO) into the paw caused a gradual increase in flinching of the injected paw and this algogenic behavior corresponded with an increase in the CSF-Glu level. 2. The nerve growth factor (NGF) inducer, 4-methyl catechol (4MC), enhanced the frequency of flinching and this effect was dose dependent. In addition, spinal CSF-Glu release was significantly above baseline 10 min after MO injection. In contrast, morphine (MOR) pretreatment completely blocked this behavioral and neurohumoral effect. 3. Anti-NGF paw injection attenuated the algogenic behavior and spinal Glu release otherwise observed after 4MC treatment. 4. The results demonstrated that MO-induced hyperalgesia is associated with increased CSF-Glu release and that this effect is potentiated by a NGF inducer. These data also suggest a possible involvement of NGF in the development of central sensitization after acute peripheral nociceptive stimulation. PMID- 10384259 TI - Distribution of 1,25-dihydroxyvitamin D3 receptor immunoreactivity in the rat olfactory system. AB - 1. The rat olfactory system contains numerous target sites for 1,25 dihydroxyvitamin D3, as determined by receptor protein (VDR) immunocytochemistry and in situ hybridization. 2. Nuclear and cytoplasmic VDR immunoreactivity as well as the corresponding hybridization signal was observed in neurons in the olfactory epithelium, the olfactory bulb, and throughout the limbic system in locations also known to be glucocorticoid targets. 3. The widespread distribution of VDR indicates the distinct functional importance of 1,25-dihydroxyvitamin D3 for olfactory perception. PMID- 10384258 TI - Glucocorticoid regulation of motoneuronal parameters in rats with spinal cord injury. AB - 1. Glucocorticoids exert beneficial effects after acute CNS injury in humans and experimental animals. To elucidate potential mechanisms of glucocorticoid action in the lesioned spinal cord, we have studied if treatment with dexamethasone (DEX) modulated the neurotrophin binding receptor p75 (p75NTR) and choline acetyltransferase (ChAT), a marker of neuronal functional viability. 2. Rats with a sham operation or with spinal cord transection at the thoracic level received vehicle or DEX several times postlesion and were sacrificed 48 hr after surgery. The lumbar region caudal to the lesion was processed for p75NTR and ChAT immunoreactivity (IR) using quantitative densitometric analysis. 3. We observed that p75NTR-IR was absent from ventral horn motoneurons of sham-operated rats, in contrast to strong staining of neuronal perikaryon in TRX rats. Administration of DEX to TRX rats had no effect on the number of neuronal cell bodies expressing p75NTR-IR but significantly increased the number and length of immunostained neuronal processes. 4. Furthermore, spinal cord transection reduced ChAT immunostaining of motoneurons by 50%, whereas DEX treatment reverted this pattern to cells with a strong immunoreaction intensity in perikaryon and cell processes. 5. It is hypothesized that increased expression of p75NTR in cell processes and of ChAT in motoneurons may be part of a mechanism by which glucocorticoids afford neuroprotection, in addition to their known antiinflammatory effects. PMID- 10384260 TI - Influence of age on stress responses to metabolic cage housing in rats. AB - 1. We studied the effect of isolation stress in 3- and 12-month-old rats individually housed in metabolic cages for 7 days. Urine (24 hr) was collected daily from one group of animals of each age. The other group was tested in an open field and on a hot plate on days 1 and 7. 2. Total deambulation in the open field test was lower in young than in older rats both on day 1 (54.7 +/- 9.9 vs 80 +/- 8.9 crossings/session; P < 0.04) and on day 7 (21 +/- 9 vs 48 +/- 7 crossings per session; P < 0.04) and decreased significantly in the two groups when tested on day 7 (P < 0.03). Latency to paw-licking in the hot-plate test was longer in young than in older animals on day 1 (14 +/- 2 vs 8 +/- 4 sec; P < 0.05) but was similar in the two groups on day 7. 3. Urinary excretions of norepinephrine (NE) and epinephrine (E) were determined by HPLC with electrochemical detection. Urinary NE in day 1 was similar in young and older animals (2627 +/- 828 vs 3069 +/- 598 ng/24 hr). In young animals NE excretion decreased along the study and was significantly (P < 0.02) lower than on day 1 during the last 3 days of the study. Conversely, in older animals urinary excretion of NE remained similar throughout the study. On day 7 urinary excretion of NE in older animals was about two fold that in young rats. Urinary E was similar in young and older rats (341 +/- 127 vs 532 +/- 256 ng/24 hr) on day 1 and showed a tendency to increase throughout the study. 4. Urinary monoamine oxidase inhibitory (IMAO) activity was determined by testing the ability of urine extracts to inhibit rat liver MAO activity in vitro and was higher in young than in older animals throughout the study (day 1, 54.8 +/- 4.2 vs 25.1 +/- 5.1%; P < 0.02). In young rats excretion of IMAO was significantly higher during the last 3 days of the study than on day 1 (P < 0.05). In older animals urinary IMAO showed a tendency to increase at the end of the study. 5. Isolation stress caused by housing rats in metabolic cages results in different behavioral and metabolic responses in young and older animals. Young animals exhibit a lower locomotor and analgesic response and excrete lower amounts of NE and higher IMAO activity in the urine than older rats. The metabolic and behavioral responses to isolation stress are highly dependent on the age of the animals tested. These results should be taken into consideration when designing experiments requiring the use of metabolic cages. PMID- 10384261 TI - Differential distribution of gonadotropin-releasing hormone variants in the brain of Hydrochaeris hydrochaeris (Mammalia, Rodentia). AB - 1. In a previous paper we reported evidence for the presence of mGnRH- and sGnRH like peptides in the preoptic-hypothalamic region of the capybara Hydrochaeris hydrochaeris (Montaner et al., 1998). In that study, the presence of a cGnRH-II like molecule in olfactory bulb extracts was suggested. 2. The capybara, the largest living rodent in the world, belongs to the order Hystricomorpha, which is considered to be one of the oldest groups of rodents. Some authors consider that this group is the ancestor of all remaining rodents. 3. In this study we have characterized GnRH molecular variants found in extracts from the olfactory bulbs and the mesencephalic region of capybara. These regions represent the two GnRH neuronal systems: the terminal nerve-septopreoptic and the midbrain systems. 4. An indirect method combining reverse-phase high-performance liquid chromatography (RP-HPLC) and radioimmunoassay (RIA) was used to characterize GnRH variants. The analysis of both extracts with two different RIA systems revealed three immunoreactive GnRH peaks, coeluting with mGnRH, cIIGnRH, and sGnRH synthetic standards. These results were additionally supported by serial dilution studies with specific antisera. 5. To our knowledge this the first report on the presence of three GnRH variants in the brain of an eutherian mammal. These results suggest that, similarly to other vertebrates, the expression of multiple GnRH variants may also be a common pattern in mammals. PMID- 10384263 TI - 5-Hydroxytryptamine inhibits P2X2 receptor channel pore mutants. AB - 1. 5-Hydroxytryptamine (10 microM) enhanced ionic current mediated through the wild-type P2X2 receptor/channel expressed in Xenopus oocytes. 2. 5 Hydroxytryptamine (10 microM) inhibited a current mediated through P2X2 receptor/channel mutants when Thr330 or Asn333 was replaced with Ile (T330I and N333I). 3. Our results suggest that neutralization of Thr330 or Asn333 exposes a high-affinity, inhibitory binding site for 5-hydroxytryptamine. This implies that 5-hydroxytryptamine interacts with the P2X2 receptor/channel at their channel pores. PMID- 10384262 TI - Adenylyl cyclase interaction with the D2 dopamine receptor family; differential coupling to Gi, Gz, and Gs. AB - 1. The D2-type dopamine receptors are thought to inhibit adenylyl cyclase (AC), via coupling to pertussis toxin (PTX)-sensitive G proteins of the Gi family. We examined whether and to what extent the various D2 receptors (D2S, D2L, D3S, D3L, and D4) couple to the PTX-insensitive G protein Gz, to produce inhibition of AC activity. 2. COS-7 cells were transiently transfected with the individual murine dopamine receptors alone, as well as together with the alpha subunit of Gz. PTX treatment was employed to inactivate endogenous alpha i, and coupling to Gi and Gz was estimated by measuring the inhibition of cAMP accumulation induced by quinpirole, in forskolin-stimulated cells. 3. D2S or D2L receptors can couple to the same extent to Gi and to Gz. The D4 dopamine receptor couples preferably to Gz, resulting in about 60% quinpirole-induced inhibition of cAMP accumulation. The D3S and D3L receptor isoforms couple slightly to Gz and result in 15 and 30% inhibition of cAMP accumulation, respectively. 4. We have demonstrated for the first time that the two D3 receptor isoforms, and not any of the other D2 receptor subtypes, also couple to Gs in both COS-7 and CHO transfected cells, in the presence of PTX. 5. Thus, the differential coupling of the D2 dopamine receptor subtypes to various G proteins may add another aspect to the diversity of dopamine receptor function. PMID- 10384264 TI - A disrupted homologue of the human CLN3 or juvenile neuronal ceroid lipofuscinosis gene in Saccharomyces cerevisiae: a model to study Batten disease. AB - 1. In order to investigate the biological function of the human CLN3 gene that is defective in Batten disease, we created a yeast strain by PCR-targeted disruption of the yeast gene (YHC3), which is a homologue of the human CLN3 gene. 2. The phenotypic characterization revealed that the yhc3 delta mutants are more sensitive to combined heat and alkaline stress than the wild-type strains as determined by inhibition of cell proliferation. 3. This suggests that the yhc3 delta mutant is a good model to investigate the biological function of human CLN3 gene in mammalian cells and to understand the pathophysiology of juvenile Batten disease. PMID- 10384265 TI - Transcription regulation by repressosome and by RNA polymerase contact. AB - The original model of repression of transcription initiation is steric interference of RNA polymerase binding to a promoter by its repressor protein bound to a DNA site that overlaps the promoter. From the results described here, we propose two other mechanisms of repressor action, both of which involve formation of higher-order DNA-multiprotein complexes. These models also explain the problem of RNA polymerase gaining access to a promoter in the condensed nucleoid in response to an inducing signal to initiate transcription. PMID- 10384266 TI - RNA polymerase-DNA interaction: structures of intermediate, open, and elongation complexes. PMID- 10384267 TI - The initiator element: a paradigm for core promoter heterogeneity within metazoan protein-coding genes. PMID- 10384268 TI - X-ray crystallographic studies of eukaryotic transcription factors. PMID- 10384269 TI - Transcription in Archaea. PMID- 10384271 TI - Transcriptional regulation by DNA structural transitions and single-stranded DNA binding proteins. PMID- 10384270 TI - Polarity of transcription on Pol II and archaeal promoters: where is the "one-way sign" and how is it read? PMID- 10384272 TI - The DPE, a conserved downstream core promoter element that is functionally analogous to the TATA box. PMID- 10384274 TI - Ten years of TFIIH. PMID- 10384273 TI - The RNA polymerase II general transcription factors: past, present, and future. PMID- 10384275 TI - Crossing the line between RNA polymerases: transcription of human snRNA genes by RNA polymerases II and III. PMID- 10384276 TI - Transcription factor IIIB: the architecture of its DNA complex, and its roles in initiation of transcription by RNA polymerase III. PMID- 10384277 TI - Strength and regulation without transcription factors: lessons from bacterial rRNA promoters. PMID- 10384278 TI - The functional and regulatory roles of sigma factors in transcription. PMID- 10384279 TI - The bacterial enhancer-binding protein NtrC as a molecular machine. PMID- 10384280 TI - Gene transcription by recruitment. PMID- 10384281 TI - Use of artificial activators to define a role for protein-protein and protein-DNA contacts in transcriptional activation. PMID- 10384282 TI - Activation and the role of reinitiation in the control of transcription by RNA polymerase II. PMID- 10384283 TI - Cofactor requirements for transcriptional activation by Sp1. PMID- 10384284 TI - Role of general and gene-specific cofactors in the regulation of eukaryotic transcription. PMID- 10384285 TI - Functional analysis of TFIID components. PMID- 10384286 TI - Mechanism and regulation of yeast RNA polymerase II transcription. PMID- 10384287 TI - Functional and structural analysis of the subunits of human transcription factor TFIID. AB - The past few years have brought many new insights concerning the structure and function of TAFII proteins. In the future, further biochemical and structural studies will no doubt lead to a greater understanding of the molecular organization of TFIID complexes. A better understanding of the function of metazoan, in particular, mammalian, TAFIIs in cell cycle progression and gene activation will, however, require the use of novel genetic techniques in addition to the biochemical analyses. PMID- 10384288 TI - Mechanisms of viral activators. AB - Adenovirus large E1A, Epstein-Barr virus Zebra, and herpes simplex virus VP16 were studied as models of animal cell transcriptional activators. Large E1A can activate transcription from a TATA box, a result that leads us to suggest that it interacts with a general transcription factor. Initial studies showed that large E1A binds directly to the TBP subunit of TFIID. However, analysis of multiple E1A and TBP mutants failed to support the significance of this in vitro interaction for the mechanism of activation. Recent studies to be reported elsewhere indicate that conserved region 3 of large E1A, which is required for its activation function, binds to one subunit of a multisubunit protein that stimulates in vitro transcription in response to large E1A and other activators. A method was developed for the rapid purification of TFIID approximately 25,000-fold to near homogeneity from a cell line engineered to express an epitope-tagged form of TBP. Purified TFIID contains 11 major TAFs ranging in mass from approximately 250 to 20 kD. Zta and VP16, but not large E1A, greatly stimulate the rate and extent of assembly of a TFIID-TFIIA complex on promoter DNA (DA complex). For VP16, this is a function of the carboxy-terminal activation subdomain. An excellent correlation was found between the ability of VP16C mutants to stimulate DA complex assembly and their ability to activate transcription in vivo. Consequently, for a subset of activation domains, DA complex assembly activity is an important component of the overall mechanism of activation. PMID- 10384290 TI - Transcription regulation, initiation, and "DNA scrunching" by T7 RNA polymerase. PMID- 10384289 TI - Cooperative assembly of RNA polymerase II transcription complexes. PMID- 10384291 TI - Structural studies of Escherichia coli RNA polymerase. PMID- 10384292 TI - Interaction of Escherichia coli sigma 70 with core RNA polymerase. PMID- 10384294 TI - Role of RNA polymerase II carboxy-terminal domain in coordinating transcription with RNA processing. PMID- 10384293 TI - The transition from initiation to elongation by RNA polymerase II. PMID- 10384295 TI - Fractions to functions: RNA polymerase II thirty years later. PMID- 10384296 TI - Antitermination by bacteriophage lambda Q protein. PMID- 10384297 TI - Structure and mechanism in transcriptional antitermination by the bacteriophage lambda N protein. PMID- 10384298 TI - Mechanistic model of the elongation complex of Escherichia coli RNA polymerase. PMID- 10384299 TI - Promoter-associated pausing in promoter architecture and postinitiation transcriptional regulation. PMID- 10384300 TI - Mechanism of promoter escape by RNA polymerase II. PMID- 10384301 TI - RNA polymerase II elongation control. PMID- 10384303 TI - The yeast RNA polymerase III transcription machinery: a paradigm for eukaryotic gene activation. PMID- 10384302 TI - HIV-1 Tat interacts with cyclin T1 to direct the P-TEFb CTD kinase complex to TAR RNA. PMID- 10384304 TI - The regulation of gene activity by histones and the histone deacetylase RPD3. PMID- 10384305 TI - Targeting Sir proteins to sites of action: a general mechanism for regulated repression. PMID- 10384306 TI - Activation and repression mechanisms in yeast. PMID- 10384307 TI - The Mad protein family links transcriptional repression to cell differentiation. PMID- 10384308 TI - Histone deacetylase directs the dominant silencing of transcription in chromatin: association with MeCP2 and the Mi-2 chromodomain SWI/SNF ATPase. PMID- 10384309 TI - Gene regulation by the yeast Ssn6-Tup1 corepressor. PMID- 10384310 TI - In vivo functions of histone acetylation/deacetylation in Tup1p repression and Gcn5p activation. PMID- 10384311 TI - Signaling to chromatin through histone modifications: how clear is the signal? PMID- 10384312 TI - Regulation of transcription by multisubunit complexes that alter nucleosome structure. PMID- 10384314 TI - Structure of the yeast histone acetyltransferase Hat1: insights into substrate specificity and implications for the Gcn5-related N-acetyltransferase superfamily. PMID- 10384313 TI - TBP-associated factors in the PCAF histone acetylase complex. PMID- 10384315 TI - The establishment of active chromatin domains. PMID- 10384316 TI - Nuclear matrix attachment regions confer long-range function upon the immunoglobulin mu enhancer. PMID- 10384317 TI - ATP-dependent remodeling of chromatin. PMID- 10384318 TI - A model for chromatin remodeling by the SWI/SNF family. PMID- 10384319 TI - SWI/SNF complex: dissection of a chromatin remodeling cycle. PMID- 10384320 TI - The SAGA of Spt proteins and transcriptional analysis in yeast: past, present, and future. PMID- 10384321 TI - Specificity of ATP-dependent chromatin remodeling at the yeast PHO5 promoter. PMID- 10384322 TI - Role of chromatin structure and distal enhancers in tissue-specific transcriptional regulation in vitro. PMID- 10384323 TI - The transcriptional basis of steroid physiology. PMID- 10384324 TI - Building transcriptional regulatory complexes: signals and surfaces. PMID- 10384325 TI - The herpes simplex virus VP16-induced complex: mechanisms of combinatorial transcriptional regulation. PMID- 10384326 TI - Structure and function of the interferon-beta enhanceosome. PMID- 10384327 TI - Autoinhibition as a transcriptional regulatory mechanism. PMID- 10384328 TI - Mechanisms of activation by CREB and CREM: phosphorylation, CBP, and a novel coactivator, ACT. PMID- 10384329 TI - Regulation of SRF activity by Rho family GTPases. PMID- 10384330 TI - Summary: three decades after sigma. PMID- 10384331 TI - [Diffuse jejuno-ileitis of Crohn's disease: a separate form of the disease?]. AB - BACKGROUND AND AIMS: Diffuse jejuno-ileitis of Crohn's disease may be a homogeneous clinical subgroup. The aim of this work was to compare the demographic and clinical data at diagnosis and the initial treatments of patients with diffuse jejuno-ileitis of Crohn's disease and to the ones without this localization. PATIENTS AND METHODS: For demographic and clinical studies, 48 (32M/16F) incident cases of diffuse jejuno-ileitis of Crohn's disease diagnosed between 1988 and 1994 in the EPIMAD register were compared with 96 (48M/48F) controls diagnosed the same year. As far as for the therapeutic management, the 48 incident cases were compared with 48 controls. RESULTS: Diffuse jejuno-ileitis constituted 3.3% of the total incident cases. Median age at diagnosis was significantly lower (20 vs 23 years, P = 0.01) and an upper digestive involvement was more frequent (56% vs 34%, P = 0.03) in patients with diffuse jejuno-ileitis. These patients were more often treated by total parenteral nutrition (43.8% vs 19.6%, P = 0.01) or azathioprine (50% vs 20.8%, P = 0.005). Azathioprine was also introduced earlier (20.7 vs 40.3 months, P = 0.009). Surgery for resection was less often required in diffuse jejuno-ileitis than in controls (65.2% vs 99.8%, P = 0.02) while more stricturoplasties were performed (52.9% vs 10%, P = 0.003); overall surgical rates did not significantly differ in the 2 groups. CONCLUSION: Our series suggest that diffuse jejuno-ileitis of Crohn's disease is a subgroup of patients characterized by a young age at diagnosis, with more frequent and earlier requirement for azathioprine. PMID- 10384332 TI - [Angina-like chest pain and exertional esophageal ph monitoring]. AB - OBJECTIVES: Spontaneous chest pain attacks are uncommon during 24-hour esophageal pH monitoring in patients suffering from angina-like chest pain suspected to be acid-related. The aim of this study was to assess the diagnostic value of exertional esophageal pH monitoring and to prove that exercise testing induces chest pain and gastro-esophageal reflux and therefore improves symptomatic correlation study. METHODS: Forty three patients suffering from angina-like chest pain underwent treadmill exercise testing during a 24-hour esophageal pH monitoring. Symptom analysis was made using the symptom-association probability described by Weusten. RESULTS: During the 24-hour pH monitoring, 10 patients (23%) had a pathologic esophageal acid exposure, 20 (46%) experienced chest pain and 3 (7%) had a symptom association probability > 95%. During the exercise testing on a treadmill, 19 patients (44%) had gastro-esophageal reflux, and 14 (32%) experienced chest pain, coinciding with a gastro-esophageal reflux in 8 (19%). After exercise testing, the symptom-association probability analysis was significantly changed in 9 patients (21%), > 95% in 6 patients (14%). CONCLUSION: Exercise testing on a treadmill induces chest pain episodes during a 24-hour esophageal pH monitoring and therefore improves symptomatic correlation study in patients suffering from angina-like chest pain. PMID- 10384333 TI - [Time required for training in colonoscopy. Prospective study of 362 consecutive tests]. AB - OBJECTIVES: To assess the time required for training in colonoscopy during one year. MATERIAL AND METHODS: All complete colonoscopies performed under general anesthesia in patients having an intact colon were included. RESULTS: The mean duration of 362 colonoscopies was 23 +/- 12 min. The time required by a senior alone was 19 +/- 9 min (n = 129). There was a significant increase in procedural time (P = 0.001) if a trainee was involved in the case (n = 68) (28 +/- 12 min) and if an assistant carried out the procedure (n = 165) alone (25 +/- 14 min) or with a trainee (27 +/- 9 min). We estimated the time which would have been required for the seniors to perform all the colonoscopies. This time was increased by 24.4% by education of trainees and self-training of assistants. CONCLUSION: The duration of colonoscopy is increased by education, the cost of which should be assessed. PMID- 10384334 TI - [Clinical value of positron emission tomography in the detection and staging of recurrent colorectal cancer]. AB - BACKGROUND: Positron emission tomography (PET) has been shown useful for the staging of patients with various carcinomas. METHODS: We have applied this technique to 54 cases of colorectal carcinoma and compared it to conventional imaging techniques. RESULTS: PET had moderately higher sensitivity and specificity than conventional techniques to detect individual lesion sites (75% vs 70.8% and 63% vs 21% respectively). It detected the same number of patients with recurrences (35/39) but overestimated disease less frequently (5 cases vs 12). PET favorably influenced therapeutic management in 17 patients, indicating different or additional surgery in 9 while avoiding surgery with curative intent or unnecessary surgery in 8. In 5 cases, erroneous information provided by PET could be corrected by conventional imaging techniques. CONCLUSION: We conclude that PET appears to provide complementary information useful for staging patients with colorectal carcinomas. It can significantly modify patients management. These data should be confirmed by a prospective study. PMID- 10384335 TI - [Matrix metalloproteases in digestive pathology]. PMID- 10384336 TI - [Contribution of telemedicine applied to digestive cancer]. AB - AIM OF THE STUDY: Telemedicine offers new possibilities for multidisciplinary care of cancer patients, allowing direct communications between different, complementary and geographically distant specialists. Thus, it is possible to form oncology committees in small hospitals where all specialties are not represented. The purpose of this study was to evaluate the medical and economic impact of visioconferences in the therapeutic management of cancer patients without access to oncology centers. MATERIALS: A telemedicine network was created in Paris between the General Surgery and Gastroenterology services of Rothschild Hospital and the services of Oncology at Saint-Antoine Hospital and Radiotherapy at Tenon Hospital. The three hospitals were connected simultaneously (multipoint) by visioconference and thus constituted a pluridisciplinary oncology committee of radiotherapy, chemotherapy and surgery. Eighty seven cases were evaluated in 27 staff conferences. In 48 cases, this consisted of re-evaluating therapeutic decisions made in surgery or gastroenterology, and in 39 cases opinions were requested by surgery (18), gastroenterology (14) or oncology departments (7). RESULTS: In only 34/87 cases therapeutic agreement was reached directly. The 53 other cases (60.9%) were debated. In fact, all 39 requests for opinion in difficult therapeutic decisions resulted in consensus. Among the 48 re evaluations, disagreement persisted in one case between the surgeon in charge of the patient and the chemotherapist. Importantly, in 13 of 48 cases (27%), the discussion modified the therapeutic protocol initially proposed. The average cost was 118 French Francs per case and per center. Total initial investment was 334,762 French Francs, but the price of some equipment has already dropped from 30 to 60%. CONCLUSION: In our study, the visioconference improved management of cancer patients for a weak working cost. PMID- 10384337 TI - [What is the status of hepatitis C virus infection in France?]. PMID- 10384338 TI - [Treatment of portal hypertension]. PMID- 10384339 TI - [Prevalence of hepatitis C virus infection in pregnant woman on the island of Reunion]. AB - AIMS: The aim of this prospective study was to assess the prevalence of anti hepatitis C virus antibodies in a population of pregnant women in Reunion. METHODS: Over a 6-month period, all blood samples of pregnant women who delivered at a hospital in the south of Reunion were tested with a third generation enzyme linked immunoassay. In addition, risk factors for hepatitis C transmission were systematically looked for. RESULTS: Among the 1,455 women tested during this period, only 2 sera were found to be positive, resulting in a prevalence of 0.14%. One of these women had a history of intravenous drug use, whereas the other had no identified risk factor. This low prevalence was found to be associated with a low frequency of risk factors of C virus infection in this population: a history of transfusion and intravenous drug use was found in 2.9% and 0.21% of cases, respectively. CONCLUSIONS: The prevalence of hepatitis C virus infection is particularly low in Reunion. This low prevalence is explained by the rarity of risk factors for hepatitis C transmission in this region which is close to the African continent and has a similar high prevalence of hepatitis B virus infection. PMID- 10384341 TI - [Hepatitis virus and hepatocellular carcinoma]. PMID- 10384342 TI - [Management of patients with cirrhosis and ascites]. PMID- 10384340 TI - [Screening of patients at risk for hepatitis C virus infection in general medicine]. AB - OBJECTIVES: The French consensus conference recommended targeted screening for hepatitis C virus infection in patients with a past history of intravenous drug use or transfusion of blood products before 1991. The aim of this study was to determine the feasibility and results of targeted screening by general practitioners. METHODS: For 2 weeks, 58 general practitioners systematically asked all their patients about a past (or current) history of intravenous drug use or transfusion of blood products before 1991. In patients who responded affirmatively, hepatitis C virus screening was proposed if it had not been performed previously. RESULTS: 8,292 patients were included. Blood transfusion and intravenous drug use were present in 383 (4.6%) and 116 (1.4%) patients respectively. Positive hepatitis C virus serology had been identified before the study, in 16 and 63 patients in these groups respectively. Tests were performed in 77% and 50% of patients with a history of blood transfusion and intravenous drug use. Systematic screening showed a positive hepatitis C virus serology in 7 and 4 patients with a history of blood transfusion and intravenous drug use (representing 2% and 15% of the tests performed during the study). Globally, 79 (88%) of the 90 patients with a positive serology had been identified before this systematic screening. CONCLUSIONS: Systematic targeted screening of hepatitis C virus infection only results in a diagnosis in a few cases. In the patients who consulted a general practitioner, 70% and 94% of cases of hepatitis C in patients with a history of blood transfusion or intravenous drug use respectively, were known before systematic screening. PMID- 10384343 TI - [Natural history of ascites]. PMID- 10384344 TI - [Ascites in patients with cirrhosis. Questions for Professor Dominique Valla]. PMID- 10384345 TI - [Sarcoidosis gastropathy: diagnosis and contribution of echo-endoscopy]. AB - We report a case of gastric sarcoidosis in a 36-year old woman. The clinical and endoscopic ultrasonography findings initially suggested linitis plastica. Subsequently the discovery of unexpected gastric and then, bronchial granulomas after bronchoscopy and bilateral ocular fixation on gallium scanning, led to the diagnosis of sarcoidosis. Endoscopic ultrasonography showed abnormal hypertrophy of the third hyperechoic layer. These findings usually suggest among the etiologies of giant gastric folds, malignant diseases and particularly linitis plastica. PMID- 10384346 TI - [Cronkhite-Canada syndrome and arsenic poisoning: fortuitous association or new etiological hypothesis?]. AB - We report a case of Cronkhite-Canada syndrome in a 66 year-old man, particular by an association with arsenism. Both arsenism and the Cronkhite-Canada syndrome are a cause of ectodermal and mucosal lesions. The persistence of physical, biological and endoscopic manifestations associated with disappearance of arsenic intoxication signs allowed us to make the diagnosis. The search of arsenic in blood, nail and hair must complete the investigations in case of acquired pseudopolyposis and ectodermal changes. PMID- 10384348 TI - [Cronkhite-Canada syndrome: a new French case]. PMID- 10384349 TI - [Acute pancreatitis complicating acute dialytic hemolysis]. PMID- 10384347 TI - [Cryoglobulinemia and hepatitis c: worsening of peripheral neuropathy after interferon alpha treatment]. AB - We report 4 cases of peripheral neuropathy induced by type II IgM kappa mixed cryoglobulinemia associated with hepatitis C virus infection. Worsening of peripheral neuropathy was observed 2 to 13 weeks after the initiation of interferon. The evolution of the hepatic and neurological diseases did not coincide, as all patients decreased or normalized transaminases with interferon. After stopping interferon, peripheral neuropathy worsened (1 case), stabilized (1 case), improved (2 cases). Low doses of interferon alpha may cause worsening of peripheral neuropathy in patients with hepatitis C virus infection associated with mixed cryoglobulinemia. PMID- 10384351 TI - Search for asymptomatic coeliac disease in patients with spongiform encephalopathy. PMID- 10384350 TI - [Thrombopenia during 5-ASA treatment (mesalazine and olsalazine)]. PMID- 10384352 TI - [Vesicular metastasis of malignant melanoma revealed by recurrent hepatic colic crises]. PMID- 10384353 TI - [Treatment by porto-systemic intrahepatic shunt for refractory hydrothorax during cirrhosis]. PMID- 10384354 TI - [Hepatic large B cell non-Hodgkin's lymphoma associated with visceral leishmaniasis in an AIDS patient]. PMID- 10384355 TI - [Recurrence and survival in cancer of the rectum: does it depend on the surgeon?]. PMID- 10384356 TI - Egos and instructions. PMID- 10384357 TI - Retrospective analysis of neurapraxia and axonotmesis injuries of select peripheral nerves of the foot and ankle and their conservative and surgical treatment (external neurolysis and neurectomy). AB - There is a paucity of scientific literature that has reviewed the conservative and surgical treatment efficacy for the management of injuries causing neurapraxia and axonotmesis. This retrospective study evaluates the clinical outcomes of certain treatments for these injuries. Twenty-seven patients fulfilled the inclusion/exclusion criteria for the study, and represented both genders and a wide variety of ages, weights, levels of education, and backgrounds. Surgical intervention resulted in a slightly better clinical outcome when compared to conservative therapies. Patients undergoing surgery for a single nerve problem improved more than those who underwent surgery when three or more nerves were involved. Failure was most often associated with: 1) multiple nerve injuries, 2) a previous history of psychopathology, and 3) application of conservative therapy without surgical intervention for single nerve injury. PMID- 10384358 TI - Subtotal calcanectomy for the treatment of large heel ulceration and calcaneal osteomyelitis in the diabetic patient. AB - Diabetic, neuropathic patients are often at risk for ulceration. Those that are temporarily or permanently limited in ambulation and restricted to a supine position are over time very susceptible to heel decubiti. Twelve patients with heel ulceration and calcaneal osteomyelitis who underwent subtotal calcanectomy are presented. Aggressive debridement of all nonviable tissue was coupled with a thorough antibiotic course in all cases. Ten of the 12 were completely healed at follow-up of 13 months. Two patients died of cardiac complications unrelated to their surgery. Postoperatively, the 10 patients were placed in an ankle-foot orthosis combined with a custom-molded shoe. All patients who were ambulatory preoperatively were able to resume their function after surgery. Subtotal calcanectomy is a relatively simple procedure to perform. In the presence of adequate vasculature, it is a good alternative to below-the-knee amputation and the accompanying sequelae. PMID- 10384359 TI - The crescentic first metatarsal basilar osteotomy for correction of metatarsus primus varus. AB - The crescentic basilar first metatarsal osteotomy has been largely abandoned by the podiatric community in recent years in favor of proximal wedge-type osteotomies for the correction of metatarsus primus varus with large intermetatarsal angles. In most cases, this was due to the inherent instability of the osteotomy and difficulties with fixation. However, the crescentic osteotomy has the ability to correct in all three planes with less shortening than the wedge-type osteotomies. New fixation techniques, such as the small cannulated screw systems, have allowed for less technical difficulty in obtaining rigid internal fixation. In this article, the authors describe the results in 29 feet (27 patients) where the crescentic osteotomy was performed along with a metatarsaphalangeal joint procedure ranging from a McBride (with or without lateral sesamoidectomy) to a phalangeal osteotomy. Preoperative intermetatarsal angles ranged from 11 degrees to 22 degrees, with an average of 18.6 degrees. Postoperative intermetatarsal angles ranged from 2.6 degrees to 8.2 degrees with an average of 5.1 degrees. The preoperative hallux abductovalgus angles ranged from 25 degrees to 38 degrees, with an average of 33.6 degrees. The postoperative hallux abductovalgus angles ranged from 4 degrees to 18 degrees with an average of 11 degrees. Complications included one hallux varus, one delayed union, and three cases of superficial cellulitis that resolved with oral antibiotic therapy. PMID- 10384360 TI - Use of polymethylmethacrylate in large osseous defects in the foot and ankle following tumor excision. AB - Foot and ankle surgeons are occasionally confronted with having to fill large defects following excision of osseous lesions. This can prove to be quite challenging to the surgeon in regards to the requirement of large amounts of autogenous, allographic, or synthetic bone graft material. The amount of time spent nonweightbearing postoperatively can be quite prolonged, and the evaluation for tumor recurrence at the graft--host interface is difficult to ascertain. Polymethylmethacrylate has been used extensively in orthopedic surgery for many years in a safe manner for total joint replacement. It has also been used to fill large defects following tumor excision (i.e., giant cell tumor) and as an alternative to bone graft. This article briefly reviews the concepts of using polymethylmethacrylate in this manner and presents the use of polymethylmethacrylate in the treatment of foot and ankle lesions with three case presentations. The authors' purpose for this paper is to simply expand on the current medical literature available regarding the use of polymethylmethacrylate in the foot and ankle and to increase the awareness of foot and ankle surgeons regarding its use as a treatment alternative. A follow-up to this article is planned to present a larger patient population, longer term follow-up, and outcomes data. PMID- 10384361 TI - High-energy bilateral talar neck fractures secondary to motocross injury. AB - The authors present a case of bilateral Hawkins type II talar neck fractures sustained during a motocross race in a 23 year old man. Due to the complexity of the injuries, open reduction with internal fixation and primary subtalar joint arthrodesis was performed bilaterally. This is one of the few cases of bilateral talar neck fractures reported in the literature in the past 15 years and one of the first utilizing open reduction and internal fixation with concomitant subtalar joint arthrodesis as a primary treatment. PMID- 10384362 TI - Chondrosarcoma of the proximal phalanx. AB - An 87-year-old male presented with a painless, large mass on the dorsum of the left foot. He reported that the mass had first appeared 10 years ago and now had become so large that he could no longer tie his shoe. The mass originated from the proximal aspect of the second digit, encompassing the second web space and distal one third of the second and third metatarsals. Surgical excision of the mass was performed and pathologic diagnosis of the specimen confirmed a grade 1 chondrosarcoma. As expected with a lower grade chondrosarcoma, the patient did not have metastasis and fully recovered. While the occurrence of chondrosarcoma is not uncommon, it rarely affects the foot. This appears to be the third case of chondrosarcoma appearing in the proximal phalanx second digit of the foot. PMID- 10384363 TI - Massive fibrolipoma of a toe. AB - Soft-tissue tumors of the toes are not particularly common; more tumors of the toes arise from the skin. The deeper tumors can be either benign or malignant and must be treated with caution until a histologic diagnosis has been made. However, malignant soft-tissue tumors of the toes are fairly rare. An unusual case of a massive benign lipoma was treated by an excisional biopsy (disarticulation). PMID- 10384364 TI - Use of topical recombinant human platelet-derived growth factor-BB (becaplermin) in healing of chronic mixed arteriovenous lower extremity diabetic ulcers. AB - Lower extremity ulcers cause significant morbidity and mortality in patients with diabetes. The primary factors that contribute to the development of this type of ulcer are peripheral neuropathy and peripheral vascular disease, which are often accompanied by infection. Lower extremity diabetic ulcers are chronic and difficult to treat, in part due to underlying pathologic conditions in individuals with diabetes that can contribute to impaired wound healing. This article reports the author's experience with treatment of chronic lower extremity ulcers of mixed etiologies with recombinant human platelet-derived growth factor- BB [rhPDGF-BB, REGRANEX (becaplermin) Gel 0.01%] in a patient with multiple risk factors including long-standing insulin-dependent type 2 diabetes. PMID- 10384366 TI - Erythromelalgia: diagnosis and classification. AB - Erythromelalgia is not a commonly recognized or diagnosed condition that affects the lower extremities. The first reported case was in 1878, when Mitchell suggested the term "erythromelalgia." This condition is characterized by a burning sensation with erythema of the involved extremity. When the extremity is lowered, or heat is applied, the pain is intensified. The application of cold or elevation of the extremity will have the opposite effect of decreasing the pain. Erythromelalgia is classified as primary or idiopathic if there is no accompanying disease process. Secondary erythromelalgia is associated commonly with myeloproliferative syndrome-related thrombocythemia, and is mostly evident in adult onset of the condition. Treatment for adults with erythromelalgia includes a single daily dose of aspirin, but children who have no associated underlying disorder find little to no relief with acetylsalicylic acid. PMID- 10384365 TI - Use of postoperative steroids to reduce pain and inflammation. AB - Postoperative injection of a steroid is used by many podiatric surgeons to reduce pain and inflammation after foot surgery. The authors present a review of the literature on postoperative steroid use from many medical specialties as well as a review of wound and bone healing. The literature indicates that using a steroid is a safe and effective means to reduce postoperative pain and edema. Studies have shown steroids to delay healing, inhibit collagen synthesis, and increase the risk of postoperative infection. No author reported a delay in wound or bone healing or increased infection rate in patients in which a steroid was used. Although there is literature to support this practice, many questions remain and further investigation is needed. PMID- 10384367 TI - Making triple arthrodesis easier and more reliable. PMID- 10384369 TI - Missense mutations in the phenylalanine hydroxylase gene (PAH) can cause accelerated proteolytic turnover of PAH enzyme: a mechanism underlying phenylketonuria. PMID- 10384370 TI - Oral phenylalanine loading profiles in symptomatic and asymptomatic gene carriers with dopa-responsive dystonia due to dominantly inherited GTP cyclohydrolase deficiency. PMID- 10384371 TI - Variant of dihydropteridine reductase deficiency without hyperphenylalaninaemia: effect of oral phenylalanine loading. PMID- 10384372 TI - Cerebrospinal fluid nitrite plus nitrate correlates with tetrahydrobiopterin concentration. PMID- 10384373 TI - Pyroglutamic aciduria and nephropathic cystinosis. PMID- 10384374 TI - Increased excretion of coproporphyrin I in a patient with hereditary tyrosinaemia type I: relevant changes with NTBC treatment. PMID- 10384375 TI - Cystinylglycinuria: a new neurometabolic disorder? PMID- 10384376 TI - Reduced glutathione, gamma-glutamylcysteine, cysteine and gamma-glutamylglutamine in gamma-glutamyltransferase deficiency. PMID- 10384377 TI - Rapid diagnosis and methionine administration: basis for a favourable outcome in a patient with methylene tetrahydrofolate reductase deficiency. PMID- 10384379 TI - Minor facial anomalies in combined methylmalonic aciduria and homocystinuria due to a defect in cobalamin metabolism. PMID- 10384378 TI - Decreased circulating plasma lipids in patients with homocystinuria. PMID- 10384380 TI - What is the origin of 3-methylglutaconic acid? PMID- 10384381 TI - Biochemistry of glutaric aciduria type I: activities of in vitro expressed wild type and mutant cDNA encoding human glutaryl-CoA dehydrogenase. PMID- 10384382 TI - 3-Hydroxyglutaric and glutaric acids are neurotoxic through NMDA receptors in vitro. PMID- 10384383 TI - Prenatal diagnosis of Canavan disease--problems and dilemmas. PMID- 10384384 TI - Identification of the molecular defect in a severe case of carnitine acylcarnitine carrier deficiency. PMID- 10384385 TI - Carnitine-acylcarnitine translocase deficiency is a treatable disease. PMID- 10384386 TI - Docosahexaenoic acid and retinal function in children with long-chain 3 hydroxyacyl-CoA dehydrogenase deficiency. PMID- 10384387 TI - DNA-based prenatal diagnosis for very-long-chain acyl-CoA dehydrogenase deficiency. PMID- 10384388 TI - Determination of total fatty acids in plasma: cis-5-tetradecenoic acid (C14:1 omega-9) in the diagnosis of long-chain fatty acid oxidation defects. PMID- 10384389 TI - Problems in the detection of fatty acid oxidation defects: experience of a quality assurance programme for qualitative urinary organic acid analysis. PMID- 10384390 TI - Adsorption of small hydroxy acids on glass: a pitfall in quantitative urinary organic acid analysis by GC-MS. PMID- 10384391 TI - Diagnosis of inborn errors of metabolism using 1H NMR spectroscopic analysis of urine. PMID- 10384392 TI - Quantitative plasma acylcarnitine analysis using electrospray tandem mass spectrometry for the diagnosis of organic acidaemias and fatty acid oxidation defects. PMID- 10384393 TI - Rapid analysis of conjugated bile acids in plasma using electrospray tandem mass spectrometry: application for selective screening of peroxisomal disorders. PMID- 10384394 TI - Mutations in PEX1 in peroxisome biogenesis disorders: G843D and a mild clinical phenotype. PMID- 10384395 TI - Disorders of peroxisome biogenesis: complementation analysis shows genetic heterogeneity with strong overrepresentation of one group (PEX1 deficiency). PMID- 10384396 TI - Molecular basis of Sjogren-Larsson syndrome: frequency of the 1297-1298 del GA and 943C-->T mutation in 29 patients. PMID- 10384397 TI - Molecular study of spinal muscular atrophy patients with hybrid genes in Bulgaria. PMID- 10384398 TI - Molecular and biochemical basis for variants and deficiency forms of galactose-1 phosphate uridyltransferase. PMID- 10384399 TI - Glycogen storage disease type IV presenting as hydrops fetalis. PMID- 10384400 TI - Cost-effective design for dental randomized clinical trials with longitudinal observations. AB - In general, randomized clinical trials (RCT) in dentistry involve longitudinal observations. In such studies, the total cost is a function of the number of study subjects and visits, the study duration, and the type and number of examinations at each visit. In this paper, we derived the minimum cost design for longitudinal RCTs with 2 treatment arms and multiple visits. We optimized the number of subjects, visits and repeated measurements under the constraints of the requirements for statistical significance, power and minimum total study cost. A SAS macro was written and made available on the World Wide Web, so interested clinical investigators can easily find optimal designs. The application of the program is illustrated using an example. PMID- 10384401 TI - Flow cytometry to monitor phagocytosis and oxidative burst of anaerobic periodontopathogenic bacteria by human polymorphonuclear leukocytes. AB - The reduced susceptibility to phagocytosis found among some periodontopathogenic anaerobes may account for the differences between invasive and non-invasive strains. We applied flow cytometry as a powerful tool to analyze and quantify phagocytosis using standardized cultures of oral anaerobes (Porphyromonas gingivalis, Prevotella intermedia, P. nigrescens, Capnocytophaga gingivalis, C. ochracea, C. sputigena, Fusobacterium nucleatum and Peptostreptococcus micros) and heparinized whole blood. Bacteria were labeled by a fluorescein-methylester and their esterase activity, resulting in green fluorescence. Ingested bacteria could be detected easily and quantified by a shift towards green fluorescence in the PMNL population involved and a concomitant decrease in the bacterial population. Furthermore, the oxidative burst of PMNLs was detected in parallel assays using the dye DHR123 which becomes fluorescent upon oxidation during the oxidative burst process. We found a great diversity in phagocytosis susceptibility determined by estimating the portion of phagocytosing PMNLs, ranging from 10.6% (strain W83) to > 99.4% (e.g. ATCC 33277T) in P. gingivalis and from 15.9% (strain MH5) to > 95% (ATCC 33563T) in P. nigrescens. In contrast, almost all P. intermedia strains as well as the representatives of the other anaerobic, putative periodontopathic species tested showed no or only moderate resistance in the phagocytosis assay. Comparison of clinical data of patients and the extent of phagocytosis resistance of the corresponding P. gingivalis strains suggests that this virulence factor may contribute to the clinical outcome. PMID- 10384402 TI - Phenytoin metabolism to 5-(4-hydroxyphenyl)-5-phenylhydantoin (HPPH) in man, cat and rat in vitro and in vivo, and susceptibility to phenytoin-induced gingival overgrowth. AB - Interspecies differences in phenytoin (PHT) metabolism to 5-(4-hydroxyphenyl)-5 phenylhydantoin (HPPH) were examined in human, cat and rat hepatic microsomes in vitro. Rat liver microsomes were 25 and 650 times more efficient at the conversion of PHT to HPPH than human and cat liver microsomes, respectively. Sulphaphenazole (83%) and tolbutamide (TOL) (64%) were the most potent inhibitors of HPPH formation in human liver microsomes, while ciprofloxacin (27%), enoxacin (27%) and TOL (26%) produced the greatest inhibition in cat liver microsomes. TOL was tested for its effect on HPPH formation and gingival overgrowth in cats in vivo. Eight cats received PHT sodium (4 mg/kg/d) and another 8 cats received PHT sodium together with TOL (20 mg/kg/d) for 10 wk. Six cats (75%) in the PHT group and 4 cats (50%) in the PHT & TOL group developed significant gingival overgrowth by the end of the study. However, the extent and incidence of the overgrowth were similar in the 2 groups. There were no significant differences in mean AUC 0-10 weeks for plasma PHT (552.90 +/- 29.6 micrograms.d/mL [PHT alone] vs. 582.41 +/- 24.49 micrograms.d/mL [PHT & TOL]) and unconjugated HPPH (1016.4 +/- 295.5 ng.d/mL [PHT alone] vs. 1174.5 +/- 397.2 ng.d/mL [PHT & TOL]) concentrations between the 2 groups of cats. Neither PHT nor HPPH were detectable in the plasma of 8 rats which received PHT (4 mg/kg/d) over a 10-wk period. The rats showed no sign of gingival inflammation (mean gingival index = 0) or gingival overgrowth (mean gingival overgrowth index = 0). Thirty-six adult epileptic patients on chronic PHT therapy were examined; 17 (47%) of the patients demonstrated clinically significant overgrowth. The mean steady-state plasma PHT concentration was comparable to, and the mean plasma unconjugated HPPH concentration 5-fold greater than, that observed in the cats. The results suggest that the rapid metabolism and elimination of PHT and HPPH in the rat may enable it to become more resistant towards developing gingival overgrowth, compared to the cat and man. PMID- 10384403 TI - Periodontal-derived cells attach to cementum attachment protein via alpha 5 beta 1 integrin. AB - A specific collagenous cementum attachment protein (CAP) has been identified in human cementum which promotes selective cell migration towards and attachment of various periodontal derived cell populations to root surfaces in vitro. The CAP is known to support attachment of periodontal-derived cell via an RGD motif, which suggests an integrin-mediated mode of attachment. The purpose of the present study was to ascertain which integrin(s) are involved in the attachment of periodontal-derived cells to CAP. The integrins examined comprised subunits of the major receptors for fibronectin (alpha 5) and collagen (alpha 2, alpha 3), as well as the common beta 1 subunit which is present in many extracellular matrix receptors. The wells of 48-well non-tissue culture treated plates were coated with CAP (2 micrograms/ml). For negative and positive controls the wells were coated with bovine serum albumin and fibronectin (5 micrograms/ml), respectively. Human gingival fibroblasts and periodontal ligament fibroblasts were labeled with [3H]-proline, incubated with anti-integrin antibodies and added to the precoated wells. Attachment was assessed after incubating the cells for 1 h at 37 degrees C in the presence of the antibodies. Antibodies to alpha 5 and beta 1 inhibited the attachment of both human gingival fibroblasts and human periodontal ligament fibroblasts to CAP, while anti alpha 2 and alpha 3 antibodies did not affect the attachment. The binding of the fibroblasts to fibronectin was also inhibited by anti-alpha 5 and beta 1 antibodies, both of which are components of the "classical" fibronectin receptor and remained unaffected by the addition of anti alpha 2 and alpha 3 antibodies. Proteins migrating in SDS-polyacrylamide gels in positions similar to the alpha 5 and beta 1 integrin subunits were present in fractions bound to a column of CAP coupled to Sepharose CL-4B. These results indicate that the attachment to CAP of the periodontal-derived cells, human gingival fibroblasts and human periodontal ligament fibroblasts, is mediated primarily via the integrin alpha 5 beta 1. PMID- 10384404 TI - Calcitonin gene-related peptide acts as a mitogen for human Gin-1 gingival fibroblasts by activating the MAP kinase signalling pathway. AB - In many peripheral tissues, calcitonin gene-related peptide (CGRP) is released from peptidergic sensory nerve fibres and acts like a growth factor during tissue development and regeneration. However, the ability of CGRP to influence gingival tissue has not been studied. To address this question, we have now examined the effects of CGRP on the proliferation of human gingival fibroblasts (Gin-1) in vitro. Gin-1 cells have approximately 3100 specific CGRP-binding sites with a Kd of 38.6 pM on their surface. Treatment with CGRP (0.1-100 nM) significantly stimulated cell proliferation in a dose-dependent manner, with maximal effects at 1-10 nM CGRP after 2 d. As one early cellular response to CGRP, p44-MAPK protein (also known as the extracellular signal response kinase [ERK]) was tyrosine- and threonine-phosphorylated within 2 min, and this phosphorylation was sustained for at least 1 h. The dose-response curve of MAPK activation was very similar to that observed for CGRP's stimulation of cell proliferation. In addition, CGRP's activation of MAPK stimulated its ability to phosphorylate the Elk-1 transcription factor. When cells were pretreated with PD98059, a selective inhibitor of MAPK kinase (also known as MEK), CGRP not only failed to induce phosphorylation of MAPK but also failed to stimulate Gin-1 cell proliferation. Our present data indicate that CGRP rapidly activates the MAPK signalling pathway, an effect which consequently stimulates the proliferation of gingival fibroblasts. Our data demonstrate specific cellular responses to CGRP by gingival fibroblasts and support the possibility that CGRP acts as a targeted local factor in the regulation of development, generation and/or regeneration of gingival tissues. PMID- 10384405 TI - Oral administration of Porphyromonas gingivalis fimbriae with cholera toxin induces anti-fimbriae serum IgG, IgM, IgA and salivary IgA antibodies. PMID- 10384406 TI - Serum antibody response against oral Eubacterium species in periodontal disease. PMID- 10384407 TI - Cybermedicine: how computing empowers patients for better health care. PMID- 10384409 TI - The electronic patient record in general practice--a South-African perspective. AB - A research project was undertaken to design an electronic medical record system which will cater for the specific needs of South African general practitioners and their unique requirements. The project was launched by sending out a questionnaire to general practitioners in the Free State, trying to determine their specific requirements and to gain insight into their attitudes and opinions regarding the use of computers, especially the computer-based patient record, in a general practice setting. The results indicated that there was a need to replace the current manual patient record with an electronic version, and that the attitudes of the general practitioners in South Africa were generally favourable regarding implementing and using such a system. Given these results, we can proceed to design a computer-based patient record system, specifically addressing the needs and requirements of the South African general practitioner. PMID- 10384408 TI - A distributed medical record on a Holter-ECG archiving and analysis system. AB - At the Department of Cardiology of the Technical University of Munich and the Deutsches Herzzentrum Munchen cardiac disease patients are treated in close co operation. In order to support the collaborative treatment as well as cardiological studies a distributed medical record (DMR) has been implemented on the basis of a Holter-ECG archiving and analysis system. Architecture, experiences and major benefits of the solution are reported in the paper. Between January 1995 and December 1997 6500 Holter-ECGs have been archived and analysed. The DMR is in routine operation since January 1996 for all heart catheter patients. PMID- 10384410 TI - The electronic patient record as a guarantor of personalised mental health care. AB - A provider of mental health services on multiple sites experienced major problems of availability with traditional paper records and commenced development of an electronic patient record (EPR) and clinical information system in order to provide integrated, real time patient based information. The development process, including clinical and technical considerata is described in order to draw conclusions about the ways in which EPR's can be a vehicle for change from traditional practice to the multidisciplinary, community based practise developments of recent years. The system can generate information for continuous quality improvement, service planning, and philosophies of care. The key requirements of good record systems are established and the inability of paper records to meet them. In contrast the electronic record has demonstrated ability to guarantee personalised, quality, mental health care. PMID- 10384411 TI - Transporting an electronic patient record system across international boundaries- the lessons learnt. AB - This paper will outline the tasks involved, completed and not achieved over an eight year period involving the implementation of the Johns Hopkins Oncology Center Information System (OCIS) in an oncology department of a secondary/tertiary care hospital in Australia. PMID- 10384412 TI - A quantitative perspective on the virtual patient record (VPR) and its realization. AB - The Virtual Patient Record (VPR), the union of all collections of health relevant data that accumulates over a person's lifetime in any institution that person has contact with, is a technical possibility in the age of networked computers[1]. This paper investigates, under what conditions the VPR may be useful and manageable. Quantitative considerations of the VPR for a population of 10 M people form the basis. Dynamic aspects of updating and decay in value for treatment decisions are also taken into account. The role of centralized and distributed data stores are compared and the need for indexing is identified. PMID- 10384413 TI - Managing healthcare: a view of tomorrow. AB - This paper presents a vision of the future in which standards exist at all levels necessary to accomplish true interoperability. The infrastructure has been established to support connectivity among all healthcare-related institutions as well as the population at large. Provider and patient care integrated in the process of an individual's care. PMID- 10384414 TI - Smartcard--a vehicle of access to the health care services. AB - This paper presents an outline of the Slovenian national project of introduction of the health insurance card system: the background, the grounds for its launching and benefits anticipated, system design, project phases, and interoperability with international healthcard systems. The system promises to simplify appreciably the administrative procedures currently required in the patient's daily encounters with the health care system, to benefit both the patient, the health care professionals and the insurance providers; as well as to provide the health care professionals with a new tool of accessing the networked information technology resources. In addition, the system cost-benefit analyses indicate significant economic benefits, which is a significant contribution to the containment of health care service. In the present environment of trans border mobility, the significance of international interoperability of any social service system is equally significant as its internal efficiency. Consequently, the issues of compliance to international technology and interoperability standards have been assigned the same high priority as the issues of technological features of the proposed solution in the system design and implementation phase. PMID- 10384415 TI - CareWeb, a web-based medical record for an integrated healthcare delivery system. AB - With the advent of Integrated Healthcare Delivery Systems, medical records are increasingly distributed across multiple institutions. Timely access to these medical records is a critical need for healthcare providers. The CareWeb project provides an architecture for World Wide Web-based retrieval of electronic medical records from heterogeneous data sources. Using Health Level 7 (HL7), web technologies and readily available software components, we consolidated the electronic records of Boston's Beth Israel and Deaconess Hospitals. We report on the creation of CareWeb (freya.bidmc.harvard.edu/careweb.htm) and propose it as a means to electronically link Integrated Health Care Delivery Systems and geographically distant information resources. PMID- 10384417 TI - Design and implementation of a multi-institution immunization registry. AB - One of every four children in the USA is underimmunized. Surveys of children in New York City have documented rates of appropriate immunization as low as 37% in certain populations in northern Manhattan. In response to this, government and private agencies have undertaken efforts to improve immunization rates. As part of one such multiinstitution effort in northern Manhattan, we have begun implementation of a computer-based immunization registry. Key features of this registry system include adaptation of legacy software in order to perform initial capture of data in electronic format; design of a user interface using a World Wide Web server that provides data review and capture functions with appropriate security; implementation of a registry database with links to the server, communication links between hospital registration systems, a Master Patient Index, community providers and the central registry; and integration of decision support in the form of Medical Logic Modules encoded in the Arden Syntax. We discuss our design of this multi-institution immunization registry and implementation efforts to date. PMID- 10384416 TI - ATRAS: a decision support application layered on the W3-EMRS architecture. AB - The creation of portable decision support systems (DSSs) remains an important goal in medical informatics. The Adrenal Test Retrieval and Analysis System (ATRAS) has been developed as an example of a simple yet highly effective decision support application which runs in a system-independent way. It is the first implementation of a decision support application layered on the World Wide Web Electronic Medical Record System (W3-EMRS) architecture, which allows for unified access to remote heterogeneous electronic medical record system (EMRS) databases. PMID- 10384418 TI - A hypermedia-based medical records management system. AB - This article presents a new way to manage computerized medical records, based on a totally-hypermedia system. As a matter of fact, the classical use of a database limits the necessary variability of the medical record, in function of both the patient profile and the care practitioner habits. The system we propose is based on a hospital Intranet, and on the XML language. This language allows the definition of semantic tags in hyperdocuments, and thus information retrieval is ensured through semantic tags indexation. PMID- 10384419 TI - A comparative study of computerised medical records usage among general practitioners in Australia and Sweden. AB - This article is based on a major empirical study of the state of adoption of Computerised Medical Records (CMRs) among General Practitioners (GPs) in Australia and Sweden. Responses were gained from a mail out questionnaire to random samples of GPs in both countries (n = 600/country). This paper will report on the main findings gained emphasising some of the various similarities and differences between the two sample groups. This comparative study adds to the existing body of CMR literature by way of providing a cross cultural perspective on GP adoption states. As a result, some concluding comments are offered for understanding high and low diffusion rates of CMRs among GPs and the implications for health policy and technology adoption strategies. PMID- 10384420 TI - Can a large institution go paperless? AB - Since 1989, Beth Israel Hospital has been deploying an extensive online patient record (the OMR), which augmented a heavily used integrated hospital information system. Initially begun in a large primary care practice, the system is now used to share patient records among 36 practices on three campuses. Although the system was intended to eliminate the need for paper, we have found that it has, in the short term, increased the amount of paper produced. Elimination of paper record in ambulatory care has saved us $56,000, but we have yet to realize the savings of an additional $200,000 per year. We explore the factors that contribute to this "paper paradox" and discuss the costs associated with increased paper production, areas in which we have reduced paper handling, and strategies for reducing our reliance on paper. PMID- 10384421 TI - User needs and demands of a computer-based patient record. AB - Prior to the implementation of a computer-based patient record, it is necessary to outline the requirements of the medical personnel. The paper is an account of a survey on information needs and demands on computer-based patient records. The study was conducted among physicians, nursing staff and therapists in two Dutch hospitals. In order to conduct the study, a measuring-instrument in form of a questionnaire was developed. Based on the results, it may be concluded, that health service staff does not only require improved input- and consultation uses with regard to the hard copy, but is also in need of additional functions. The developed measuring instrument appeared to be a proficient aid in outlining the information needs and demands of the health service staff. Through the developed questionnaire, the staff was able to obtain an idea of the possibilities of the computer-based patient record and state their own interest in same. PMID- 10384422 TI - A multimedia database for dermatology. AB - The World Health Organisation (WHO) Radiation Emergency Medical Preparedness (REMPAN) centres have built up the International Computer Database for Radiation Accident Case Histories (ICDREC) to document the treatment of acute radiation syndrome (ARS) patients. Radiation induced skin lesions may cause severe late effects in radiation accident patients. Dermatological multimedia documentation is included into the ICDREC. In particular, retrieval and display of digitised skin photographs and medical reports serves to improve patient care, medical education, and scientific analysis concerning the cutaneous radiation syndrome (CRS). The database has been built up as a client/server system. A particular focus has been set on using commercial off-the-shelf software components. The medical data including the multimedia data are stored in a relational database system. The database can be accessed by inexpensive personal computers in the dermatologist's workplace. Authorised institutions can access the database via the Internet. Retrieval of one skin photograph via local area network (LAN) requires approximately 3 seconds. The current state of the application is illustrated with the skin lesion treatment of a Chernobyl patient. An example is given on how to access the ICDREC from a dermatologist's desktop personal computer. The discussion focuses on the advantages of storing the textual and pictorial data in one central database to be accessed from different care centres and how the results can be generalised for medical multimedia information systems. PMID- 10384423 TI - The survey on the completeness of the medical records as the basis for producing valuable health information. AB - The core of the health information system in the hospitals lies in the medical records, which contain all the data concerning diseases and practices. Then questions arise whether the medical records contain all the data needed in the reliable, complete and timely manners while meeting standards for confidentiality. In this study, we reviewed medical records of 11 general tertiary care hospitals in Seoul, Korea, according to the criteria we made based on the JCAHO's hospital accreditation manual. The focus of review was whether the medical records contain the valuable information fully and in timely manners. But the result was no better than our expectations. More caution should be given for the EPR software engineers to catch up all the information needed from the medical records. We also examine the cause of variation among hospitals and want to give basic information concerning the medical records for implementing the standardized EPR and suggest the method for keeping complete health information. PMID- 10384424 TI - Using a computer-based patient record for quality management in surgery. AB - Our first generation hospital information system is in routine use since 1989. The patient data are used for multiple clinical purposes: operation scheduling, reminder for patient care, case retrieval for research and education, and so on. The computer-based patient record was complemented by a flexible PC-based query and report facility for quality management. A regular export of patient data from the host is translated into a relational database model and assessed via the standard of Open Database Connectivity (ODBC) by a statistical analysis tool. Herewith, descriptive and analytical statistics become available to support the clinical departments in patient care. Concerning the migration of first generation systems to modern architectures the presented approach has to be compared with strategies of replacement and capsulation. A decision on the strategy applied should take into account local resources and opportunities. PMID- 10384425 TI - Devices for structured data entry in electronic patient record. AB - Electronic patient record is expected to have edit, data analysis, and decision supporting functions. To realize these functions, the entered data should be structured. We made a template based data entry system with some devices. We defined a template for each describing unit, i.e., symptom, physical finding and examination report. Template is composed of several describing elements (a pair of property and value), which form tree structure in general. When a template is selected, the top layers of the elements are displayed at once allowing data entry. When the data qualified by other elements is entered then system presents the second layer about this data at once. This enables users to skip entering some unnecessary items. Users can constitute a form by combining some templates frequently used in a situation. Furthermore, at the second patient visit, the system can present the templates used in the former patient visit to check the different point. These templates and forms can be made easily by editing the master data using template master maintenance program. The entered patient data are presented in progress note and flow sheet. In progress note, the entered data are translated into natural language. In the flow sheet, representative data of each template are present in the cell of the matrix whose line indicates the describing unit and column indicates date. If the cell is clicked, then the details are presented. Using this system, we made templates and forms for cardiovascular field and entered the data about an actual patient with angina pectoris. The time taken by inputting data is shorter than that by handwriting and the content is enough for a patient record. This system is practical for structured data entry in electrical patient record. PMID- 10384426 TI - On stepwise integrating an electronic patient record based on digital-optical archiving and multi-purpose health professional workstations: experiences at the Heidelberg University Hospital. AB - At the Heidelberg University Hospital the conventional paper-based medical record is currently being replaced by a "unique" electronic patient record (EPR). This paper describes the stepwise integration of an EPR based on digital-optical archiving and multi-purpose health professional workstations. If focuses on the potentials of digital optical archiving as an integral part of hospital information systems. So far, the EPR has been introduced in the central archive of the "head clinic", the Neurosurgical Clinic and the inpatient archive, the endoscopy and sonography section of the Department of Internal Medicine. PMID- 10384427 TI - Data collection in multi-center clinical trials via Internet. A generic system in Java. AB - Data collection via Internet is usually performed with an HTML/CGI combination, which has a lot of disadvantages, most important the lack of security features. We therefore have developed a system written entirely in Java, which implements a true client/server application based on TCP/IP. The documents are created using a multi-lingual data dictionary, and the used GUI components are able to perform plausibility checks, which improves quality of the data. The system is designed to be easily extensible so that it can be used in almost any kind of clinical trials. It is based on a three-tier model where client requests are handled and monitored by an application server. We will describe this system and it's implementation and compare it to the HTML/CGI approach. Of special interest are security features, which are possible through the use of Java. PMID- 10384428 TI - Design and development of a computer-based clinical referral system for use within a physician hospital organization. AB - The process of creating a clinical referral for a patient and the transfer of information from the primary care physician to the specialist and back again is a key component in the struggle to deliver less costly and more effective clinical care. We have created a computer-based clinical referral application which facilitates 1) identifying an appropriate specialist; 2) collecting the clinical, demographic, and financial data required to generate a referral; and 3) transferring the information between the specialist and the primary care physician. Preliminary results indicate that the new computer-based process is faster. PMID- 10384429 TI - The federated healthcare record to support shared diabetes care. AB - In this paper the conception of the federated healthcare record server to support shared diabetes care is described. Business process modelling is applied to describe the shared care for diabetes patients. Typical dialogues between the different users (patient, internist, GPs, and diabetic nurses) are analysed and described in terms of use cases. Next to this modelling three incremental steps are defined to realise the record server based upon results of standardisation. It proves to be successful to design and build this record server on modern technologies like CORBA and JAVA. PMID- 10384430 TI - New concept of an integrated IT&T-based dental workstation for quality assurance in oral health care. AB - During the last five years, quality development has become a most important issue in oral health care. In every day practice, quality development meets, however, a considerable number of objective obstacles, mainly due to the lack of direct access to the information necessary in a specific clinical work situation and due to the absence of knowledge on own performance in relation to quality goals. The EU-TAP project ORQUEST has identified different clinical work situations and built up an integrated IT&T-platform composed of different software modules and hardware components for each clinical work situation in order to allow for adequate IT&T-support in specific clinical situations. PMID- 10384431 TI - A mobile system for recording examination data of the analysis of functional disorders of the masticatory system. AB - Simplifying and streamlining data access and recording tasks in medical settings is an important and successful application area for mobile computing technology. An interesting research topic in this area are user-interface design concepts that allow for an optimal integration of system operation into working situations where the user is tightly involved in interactions with his physical surroundings. We describe an ongoing project that aims at developing a mobile information system which uses pen-computers for recording data of the analysis of functional disorders of the masticatory system during examination. Long-term goal of this project is the design of an interaction concept that allows for an optimal integration of the system operation into the established examination procedures. A comprehensive questionnaire for the analysis of functional disorders of the masticatory system has been developed. This questionnaire then has been structured with respect to established examination procedures and coded into a pen-computer. Experiences with a first system prototype within the scope of a limited field trial show that our approach is viable and simplifies the recording task. Future work will concentrate on a further streamlining of the user interface by providing additional task-specific graphics interaction techniques and by a detailed study of usage patterns. PMID- 10384432 TI - A support system to make care plan of elders. AB - The population of persons over the age of 65 is rapidly growing. In order to provide care to residents of long term care facilities and care to senior citizens in home, Minimum Data Set (MDS), the Clients Assessment Protocols (CAPs) and MDS-Home Care CAPs (MDS-HC CAPs) have been are offered by the HCFA and interRAI Overview Committee. According to offered guidelines, nursing staff spend much time to process measuring ADL (Activities Daily of Livings) of clients and to find problem areas. We have developed the information system to assist nursing staff to reduce this paper work. This system consists of generating problem areas; providing shared files to nursing staff, simple MDS input methods and mobile handy terminal which includes a video camera tool. In this paper we describe the design and implementation of Integrated Information System To Help Nursing Senior Citizens. PMID- 10384433 TI - A multifunctional system of the national genetic register. AB - The National (Federal) Russian Genetic Register operates as a multifunctional system. It is designed for finding solution to the following key problems: 1. Information provision to watch families having children with hereditary and congenital diseases. 2. Follow up of the diagnostic process (including that associated with molecular-cytogenetic studies) and making predictive decisions concerning a risk of hereditary disease in the family. 3. Monitoring of new cases of congenital/hereditary disease under the effect of genotoxic and general toxic environmental factors. 4. Rendering consulting services to those concerned and reference information on the organizational/methodological problems. The relational database is composed of textual and graphical data. This enables the doctors to enter and review the pedigree in the form they have got accustomed to. Numerous built-in classification schemes help simplify and speed up the completion of the medical records. An additional information can be entered in the textual form. When requested by the user, the information about the patient and his (her) family, stored in the database, can be displayed as a summary document (a statement). To do so, a special algorithm based on the results of the logical information analysis, as well as medical history of the patients themselves and their families' members has been put forward. The document thus compiled is open for being edited by the doctor. The local computer network is operated in the "user-server" mode. For information protection purposes, an authorized access system has been devised to protect various data categories. The software tools operate in the Windows environment for IBM-compatible personal computers. Also, use is made of Microsoft SQL-Server, Visual Fox-Pro 3.0, Visual C+2 2.1, and FORTRAN 5.1. The doctor decision modules are activated with mathematical models, an expert system, and a self-learning recognizing system. The Windows NT system is a choice for the existing Federal and Regional Genetic Centers. For territorial consulting purposes other alternatives might be Windows for Workgroup 3.11 or Windows 95. The local computer networks in individual organizations will be integrated to form a Corporate Medical Genetics Service of Russia. The system is to be completed before the year 2000 in a number of phases. Up to now, program packages for family database management and recurrent risk calculations have been worked out and tested. PMID- 10384434 TI - Nursing e-journals: are current models using the web's potential? AB - As the number of nursing and health-related electronic journals (e-journals) available on the World Wide Web increases, we are seeing an increasing diversity of models. A taxonomy of current nursing e-journal models is presented, together with brief discussion of some of the issues pertinent to the development of e journals, and that currently appear to constrain the electronic publication of high quality content. We describe in detail the model and usage of one particular e-journal, Nursing Standard Online. We conclude by advocating the development of innovative and increasingly interactive nursing e-journals as the way forward, discussing one particular model which holds promise. We believe it likely that nursing e-journals using current models will need to be specialist rather than generalist if they are to attract a larger audience. PMID- 10384435 TI - An analysis of 5 years of medical informatics publications. AB - This study describes an analysis of 1520 papers that were published in six medical informatics journals from 1992 through 1996, and indexed in NLM's MEDLINE. Of retrieved publications the countries of origin were determined, and insight in the subject of the publication was obtained by analysing the used MeSH terms. The main conclusion is that despite the fact that publications in medical informatics stem from a wide international community, scientific recognition can still not be demonstrated by high impact factors of journals. PMID- 10384436 TI - SGML-based construction and automatic organization of comprehensive medical textbook on the Internet. AB - The amount of knowledge required in practical medicine is large and ever increasing. Medical staff must select and use appropriate pieces of the knowledge from this flood of medical information. Recent Internet technology may be solving these problems because it makes information open to the public immediately after it is created and enables many people to share it. Medical resources on the Internet are however currently not always well organized, because these are often voluntarily provided by the experts of a particular field. We therefore decided to create a comprehensive medical database on the Internet, which is well organized, and of a high quality for practical medical use. In order to make full use of the benefits provided by electronic media, we created a new structured data set of information. We then commissioned authors to write manuscripts from which we created Standard General Mark-up Language (SGML) documents. We then wrote a translation program that took the SGML and automatically created a fully inter-linked HyperText Mark-up Language (HTML) document. The translation program generated 4,814 HTML files created from 1,373 number of SGML documents. The total data size including pictures was about 640 MB. 205,775 related links were created. We then published our electronic medical textbook described in HTML publicly on the Internet. Using SGML-based structured data, we constructed a complex electronic medical textbook created organically from simple SGML instances. Our electronic medical textbook is systematic and comprehensive, and has a homogeneous structure. We believe that this is the first comprehensive medical textbook available on the Internet. Furthermore, it was found that our approach to the electronic medical textbook has two major advantages. One is automatic generation of inter-links among documents, and another is easy to maintain documents. In addition, once we construct the electronic textbase in SGML format, the data can be utilized to various application programs on different platforms. Making use of this feature, we are now planning to develop a new style of electronic textbook, which is closely integrated with a Hospital Information System (HIS). The plan would provide medical staff with on-demand access to the electronic medical textbook while using HIS terminals. PMID- 10384437 TI - A multi-agent softbot to retrieve medical information on Internet. AB - World-Wide Web is a media where information is unstructured, distributed, multimedia and multilingual. Many tools have been developed to help users search for information: subject hierarchies, general search engines, browsers and search assistants. Although helpful, they show serious limitations, mainly in terms of precision, multilingual indexing and distribution. The M.A.R.V.I.N. project (Multi Agent Retrieval Vagabond on Information Networks) proposes solutions with a distribution of the indexing task in agents specialized in a given domain. M.A.R.V.I.N. has been successfully applied to the medical domain. The medical index and its associated search engine MedHunt (Medical Hunter) demonstrate the interest of such an approach with currently 50,000 indexed medical sites and more than 100,000 accesses each month. PMID- 10384438 TI - Full text multilingual automatic morphosemantems for stand-alone or Internet based applications. AB - The authors present an automatic tool able to provide real-time morphosemantic decomposition of natural language sentences in French, German and English. This tool demonstrates the feasibility of Natural Language Processing on standard PC computers and the technology involved has been successfully implemented in daily used applications in several European hospitals. It considerably alleviates the burden of coding with various international classification and enhances the quality of the final results. This tool, delivered on PC platforms, is highly convivial and provides a versatile interface to any existing applications based on the Microsoft Windows standards. Moreover, all high levels functions have been encapsulated in Object Oriented Components and can therefore be reused using the Common Object Model standards to develop stand-alone or Internet applications. PMID- 10384439 TI - Implementation of a database on drugs into a university hospital Intranet. AB - Several databases on drugs have been developed worldwide for drug information functions whose sources are now electronically available. Our objective was to implement one of them in our University hospitals information system. Theriaque is a database which contains information on all the drugs available in France. Before its implementation we modeled its content (chemical classes, active components, excipients, indications, contra-indications, side effects, and so on) following an object-oriented method. From this model we designed dynamic HTML pages according to the Microsoft's Internet Database Connector (IDC) technics. This allowed a fast implementation and does not imply to port a client application on the thousands of workstations over the network of the University hospitals. This interface provides end-users with an easy-to-use and natural way to access information related to drugs in an Intranet environment. PMID- 10384440 TI - Conceptual integration of information databases into an Intranet. AB - Large information systems handle massive volume of data stored in heterogeneous sources of information. Each server has its own model of concepts representation with regard to its aims. One of the main problems encountered by end-users when accessing different servers is to match their own viewpoint on biomedical concepts with their various representations that are made in the database servers. The aim of the project ARIANE is to provide end-users with easy-to-use and natural means to access and query heterogeneous information databases. The objectives of this research work consist in building a conceptual interface by means of the Internet technology inside an enterprise Intranet, and to propose a method to realize it. Moreover, this method provides designers of web sites with a powerful tool to manage them on the basis of an ontology of the biomedical domain. This method is based on the knowledge sources provided by the Unified Medical Language System project of the U.S. National Library of Medicine and exploits intensively the conceptual graphs theory. PMID- 10384441 TI - UMLS-based access to CPR data. AB - This paper describes the results of a project that explores the use the Unified Medical Language System (UMLS) for knowledge-driven tasks, such as browsing a computer-based patient record (CPR). The project consisted of a number of steps: the mapping between CPR terms and UMLS concepts, the development of an algorithm that explores the CPR data using this mapping, and the implementation of a first prototype browser that visualizes "found" data. A second task addressed in this project has been the direct access to online medical literature (MEDLINE) using the UMLS concepts found in the CPR data. In this project, we used a preliminary version of the Open Records for Patient Care (ORCA) CPR that consisted only of the history and physical examination data of patient suffering from heart failure. PMID- 10384442 TI - A structured model for evaluation of information retrieval (IR). AB - The paper reviews methods of evaluation of information retrieval (IR), in particular relevance (recall and precision) based approaches and their limitations and proposes a new model of IR which emphasizes the difference between the IR process and the technical IR system supporting the process. The model is proposed as basis for the structured evaluation of the IR process, and the planning, analysis and comparison of results of experiments using different methodologies. PMID- 10384443 TI - Integration of the analytical and alphabetical ICD10 in a coding help system. Proposal of a theoretical model for the ICD representation. AB - In French hospitals, medical diagnosis coding with the ICD10 is commonly performed and the use of effective tools would help coders in their task. AIM OF THIS WORK: To ameliorate an existing coding help system. This system, which already consists of the ICD10 analytical index, would be increased with the terms of the alphabetical index that includes lexical variants and additional terms as well. The addition of the second volume of the ICD would allow the coding of more terms and would lessen documentary silence. METHODS: The first step of this work was a careful study of a theoretical model of the ICD content. Then the alphabetical index file was submitted to a lexical analysis, and it was automatically transformed to be integrated into the existing coding help system. RESULTS: Compromise had to be made between a theoretical model and between what could be obtained in practice by an automatic processing of the file. Finally the alphabetical index was added to the initial thesaurus, which represents 42,000 terms and 4,000 additional words. Links between words and codes were also considerably increased, which has enhanced the searching possibilities of the tool and lessen documentary silence. Conversely the research time has been increased. CONCLUSION: Difficulties have to be encountered when trying to turn a manual tool into an automatic research tool. PMID- 10384444 TI - The Asia Pacific Association for Medical Informatics (APAMI) and World Organisation of Family Doctors (WONCA) Consortium on General and Family Practice Informatics--a statement of intent. AB - This paper describes the establishment of a consortium to advance health and medical informatics in general/family practice in the Asia Pacific Region. The objectives, current activities currently taking place in the region and key activities planned will be outlined. PMID- 10384445 TI - An integrated multimedia medical information network system. AB - An integrated multimedia medical information network system at Shimane Medical university has been developed to organize medical information generated from each section and provide information services useful for education, research and clinical practice. The report describes the outline of our system. It is designed to serve as a distributed database for electronic medical records and images. We are developing the MML engine that is to be linked to the world wide web (WWW) network system. To the users, this system will present an integrated multimedia representation of the patient records, providing access to both the image and text-based data required for an effective clinical decision making and medical education. PMID- 10384446 TI - A global Intranet for an international scientific society. AB - The International Medical Informatics Association (IMIA) has built up a world wide infrastructure which is using the Internet as a backbone for a global Intranet. The work has been supported by cost analysis and user acceptance monitoring. As one of the first international scientific societies, IMIA is offering professional electronic communication services to its members. This step has been taken to advance international cooperation and to support the dissemination and exchange of information on the Health Informatics Sector. The current ways of communication and Information Exchange do not meet the requirements of the Information Society because they are too slow and too expensive. The implementation of an Intranet based on the Internet provides a communication channel which is easily accessible for all IMIA members and which will allow efficient information exchange and built up links between IMIA related projects and organizations. PMID- 10384447 TI - The software challenge for the next decade: the global objects scenario. AB - This paper discusses the advantages of developing software as pattern-based components. The design and implementation of a pattern-based suite of software components specially constructed for the electronic patient record is presented. The methodology and the lessons learned in the development of these components are discussed. Finally, some comments about the globalization and the need for more integration among component developers in a worldwide basis is discussed. PMID- 10384448 TI - Design and application of a terminology management system. AB - A Swedish data model for handling terminology, Spriterm, is presented in this paper. A prototype terminology management system, using the Spriterm data model in also described. This prototype is implemented is Microsoft ACCESS. Furthermore, two other applications using this prototype as a base are introduced. One World Wide Web based application, and a data dictionary. PMID- 10384449 TI - Middleware for healthcare information systems. AB - Middleware is now a commonly used expression and anyone building distributed applications is referring to "middleware services". Nevertheless this notion lacks of sound theoretical foundation. This paper tries to clarify the relationship between the components of distributed environments, especially in healthcare, and to establish some classification aiming at gaining a common understanding of the functionalities and interdependency of the existing modules of distributed environments. PMID- 10384450 TI - Object technology: raising the standards for healthcare information systems. AB - Netscape and the public Internet have accelerated the acceptance of many different open "Internet standards". Through wide acceptance of its browser, Netscape gave a boost to the Java programming language helping it become truly platform independent. Objects written in Java are ideal building blocks for application components. CORBA gives such objects the ability to communicate and operate over networks. Applications built with these distributed objects become the services in an Internet-wide healthcare framework. The convergence of object technologies has raised the standards for modern healthcare information systems. To illustrate the relationship among such technologies, this paper presents an architecture for a Universal Healthcare Information System (UHIS) in terms of its web, Java and CORBA components. PMID- 10384451 TI - From legacy systems via client/server to web browser technology in hospital informatics in Finland. AB - The majority of hospital information system installations in Finland are based on a legacy technology from the U.S. Department of Veterans Affairs (VA). This paper presents an architecture and a tool set which provide a migration path from terminal-based to client/server technology, conserving much of the investments in existing applications. It is argued, though, that a new technological revolution is required in the form of extending the web browser/server technology to operational information systems in hospitals. A blueprint is presented for a further migration path from client/server to browser/server technology. The browser technology is regarded as a major challenge to hospital information systems in the next few years. PMID- 10384452 TI - Dynamic workflow model for complex activity in intensive care unit. AB - Cooperation is very important in Medical care, especially in the Intensive Care Unit (ICU) where the difficulties increase which is due to the urgency of the work. Workflow systems are considered as well adapted to modelize productive work in business process. We aim at introducing this approach in the Health Care domain. We have proposed a conversation-based Workflow in order to modelize the therapeutics plan in the ICU [1]. But in such a complex field, the flexibility of the workflow system is essential for the system to be usable. In this paper, we focus on the main points used to increase the dynamicity. We report on affecting roles, highlighting information, and controlling the system We propose some solutions and describe our prototype in the ICU. PMID- 10384453 TI - Intelligent agent for collaborative diagnosis. AB - An agent-based approach to facilitating cooperative medical diagnosis is presented in this paper. Background work in computer supported cooperative work and medical informatics is first discussed. Relevant theory for interaction management is then considered. An agent-based interaction is then shown via a case study to facilitate cooperative diagnosis. This is achieved through monitoring patient record construction and by highlighting relevant diagnostic information. PMID- 10384454 TI - SeCD electronic folder: CADMIO's application for the medical folder of a service for the care of drug addicts. AB - In this paper we will describe the SeCD (Service for the Care of Drug addicts) electronic folder, a specific application of CADMIO [1] (Computer Aided Design for Medical Information Objects) system. CADMIO is a system for the definition, construction and management of multimedia clinical folders. The Ser.T. (Servizio per la Tossicodipendenza/Service for Drug Addicts) has earned a very special place within the Italian clinical structures as well as any service for drug addicts has done in the rest of the world. Such a structure has special needs and the characteristics of its medical folders are very different from any other folder. Actually, a Ser.T. has to keep updated the patient situation either from the clinical point of view as well as the psychiatric one. Moreover, it must keep track of the clinician subjective considerations about the patient psychic state and his situation in regard of the law. So, we had to redesign some of the features of the existing CADMIO application, to accommodate such highly not structured data into objects easily manipulated by an informative system. The objectives we hope to achieve were mainly two: To show that a well designed adaptive system can be easily exploited to support even very complex and poorly structured data types and actions To design data structures able to accommodate medical, psychiatric and administrative data in an homogeneous manner. PMID- 10384455 TI - The creation of a global telemedical information society. AB - Healthcare is a major candidate for improvement in any vision of the kinds of "information highways" and "information societies" that are now being visualized. The medical information management market is one of the largest and fastest growing segments of the healthcare device industry. The expected revenue by the year 2000 is US$21 billion. Telemedicine currently accounts for only a small segment but is expanding rapidly. In the United States more than 60% of federal telemedicine projects were initiated in the last two years. The concept of telemedicine captures much of what is developing in terms of technology implementations, especially if it is combined with the growth of the Internet and World Wide Web (WWW). It is foreseen that the World Wide Web (WWW) will become the most important communication medium of any future information society. If the development of such a society is to be on a global scale it should not be allowed to develop in an ad hoc manner. The Euromed Project has identified 20 building blocks resulting in 39 steps requiring multi-disciplinary collaborations. Since, the organization of information is therefore critical especially when concerning healthcare the Euromed Project has also introduced a new (global) standard called "Virtual Medical Worlds" which provides the potential to organize existing medical information and provide the foundations for its integration into future forms of medical information systems. Virtual Medical Worlds, based on 3D reconstructed medical models, utilizes the WWW as a navigational medium to remotely access multimedia medical information systems. The visualisation and manipulation of hyper-graphical 3D "body/organ" templates and patient-specific 3D/4D/and VR models is an attempt to define an information infrastructure in an emerging WWW-based telemedical information society. PMID- 10384456 TI - The evolution of a German teleradiology system. AB - This paper describes the evolution of a german teleradiology system. The development started from simple image file transfer, continued with a dedicated teleradiology system and ended up with a general radiology workstation with teleradiology features. The main features, advantages and drawbacks of the different generations are described. The own developments are compared with developments at other places. The influence by standards is also included in this investigation. The latest systems are mainly used by the radiologists and the image transfer for scientific cooperation is nowadays just one of several application fields of teleradiology. PMID- 10384457 TI - Telemedicine: responsibilities and contractual framework. AB - The rapid growth of telemedicine has created a need for a definition of the responsibilities of the doctors involved. These responsibilities must be analyzed according the tort of negligence as a function of the level of competence of each doctor, their unequal access to the relevant information and their command of the telemedicine system. This analysis leads on to a study of the legal value of the electronic records kept and the ways in which the doctors are remunerated. PMID- 10384458 TI - Home telecare system integrated with periodic health reminder and medical record & multimedia health information. AB - The necessity of home telecare system is growing due to increase in desire for health promotion owing to increase in chronic diseases, aged population and medical expenses. Already, we computerized patient's data and offer periodic health reminder to patients for health promotion by using Life-time Health Monitoring Program (LHMP). Our study connected LHMP to the Web on internet by CGI as an electronic medical record; enabling reference to patient's medical records anywhere. The study also made possible video teleconsultation and constructed multimedia database to provide health-related information to the patients. On these bases, a flow chart was developed using the home telecare to practice manage patients with chronic diseases, old patients, and the handicapped. Further standardization in data, establishment of law bases for home telecare system, development of rules for medical fees and active utilization of biomedical telemetry will be needed to extend home telecare system. PMID- 10384459 TI - A study on development of a home health care support information system. AB - As the need for home health care has been increasing with the rising number of the elderly in Japan, the application of medical informatics to home health care delivery is considered to be useful. Therefore, development of a home health care support information system was planned. The system can collect patient's PHD (Personal Health Data) such as data of ECG, complaints, etc. at patient's home and can send the PHD to medical facilities. We designed and constructed two subsystems on a trial basis. One subsystem has function of gathering, recording and transmitting vital signs of the aged such as ECG, physical activity rate, oxygen saturation rate in arterial blood. Another subsystem can collect and send image data of the old people at their home. Experiments for trial use of the system was conducted and it was recognized that the PHD can be smoothly collected and recorded at home of the elderly and can be sent to the medical facilities with good success by using the system. PMID- 10384460 TI - Telematic system for monitoring of asthma severity in patients' homes. AB - Despite advances in the treatment of asthma the morbidity and mortality of this disease has increased significantly in the past several years. Recent studies have shown that monitoring of asthma severity in the patient home especially combined with patient education can reduce incidence of asthma exacerbation and subsequent hospitalization. The existing methods for home asthma monitoring are limited by four factors; they completely rely on a patient's ability to document and to evaluate test results; there is no easy way for a physician to review data in a timely manner; they use imprecise tools for evaluation of asthma severity and they don't provide clinical decision support tools. The goal of this study is to develop and to evaluate a telematic system for asthma severity monitoring which will minimize patients' efforts in performing self-testing at their homes and allow prompt reciprocal exchange of all relevant information between patients and health care providers. In our setting, patients use portable spirometer and pocket-sized palmtop computer for data exchange. Our system allows daily serial monitoring of asthma severity at patients' homes using Forced Vital Capacity test and symptom diary. The results of the tests become available for physicians review immediately after completion of self-testing procedures via Web browser. The results can be transmitted from patients' homes (or any other remote location) to the medical records database via landline or wireless networks in several minutes. Each time the remote server receives patient's results, it invokes the application which tests the validity of data, analyzes parameters trends and dispatches corresponding messages for the patient and, if necessary, for physicians. Such an approach provides constant feedback loop between asthma patient and physician. The system has been tested in 10 healthy volunteers and asthma patients. Patients participated in the study from two to 21 days. The test results showed that the system provides reliable reciprocal exchange of all relevant information between a physician and asthma patient in home settings. Average transmission time from the patient's palmtop to the remote central data repository was about 1 minute for 14.4 Kbps landline modem, 6 minutes for cellular network and 8 minutes for RAM Mobile network. After transmission, the test results were immediately available for review at our web site. PMID- 10384461 TI - Ambulatory physical activity monitoring system. AB - In this study, we have developed an ambulatory behaviour map and physical activity monitoring system by equipping our portable digital biosignal memory device developed previously with a GPS sensor and piezoresistive accelerometers. By this system, we can get the subjects behaviour map, his physical activities and posture changes in daily life. PMID- 10384462 TI - Development of a medical record and radiographic image transmission system using a high-speed communication network. AB - A medical record and radiographic image transmission system has been developed using a high-speed communication network. The databases are designed to store and transmit the data acquired from the scanner. To maximally utilize the communication bandwidth, the medical records and radiographic images are compressed using the G3 facsimile and JPEG coding standard method, respectively. TCP/IP, OOP and Windows-based system software enable a modular design, future expandability, open system interconnectivity, and diverse image manipulation functions. PMID- 10384463 TI - World wide microscope: new concept of internet telepathology microscope and implementation of the prototype. AB - We defined a new concept of microscope system for telepathology, named World Wide Microscope (WWM), and implemented its prototype. WWM is constructed by the following three units; (1) microscope unit, (2) control unit, and (3) internet unit. The microscope unit is a conventional light microscope equipped with a motor drive X-Y stage and an objective lens revolver, auto focus and auto iris functions, and a CCD camera. The internet unit is a World Wide Web homepage in which a Java applet and a communication server are installed. The applet is implemented several methods that realize to make a PC client on the Internet as a telepathology terminal. The control unit relays request commands generated from the applet to the microscope unit, and captures the microscopic images. We think WWM will probably become the all round telepathology tool of the next generation. PMID- 10384464 TI - The effect of a teledermatology program on rural referral patterns to dermatologists and the management of skin disease. AB - Teledermatology can be defined as the use of imaging and telecommunications technologies to provide skin care services at a distance. The potential value of teledermatology is especially great in rural and medically underserved areas that do not have, or cannot support, providers specializing in the diagnosis and management of skin diseases. Rural patients and primary care providers should be able to use teledermatology as a greatly simplified and potentially less expensive means of referral to an urban dermatologist. In an effort to gauge the impact of a simple teledermatology system on referral patterns and the management of rural patients with skin disorders, we studied baseline rates of referral to dermatologists from five primary care clinics in rural Oregon. Economical, easy to-use teledermatology systems were subsequently installed, and the effects on patient referral and management were recorded over time. The interim results suggest that primary care providers (PCPs) are reluctant to refer patients with skin conditions, even when the primary care providers confidence in the correct diagnosis and treatment plan for that condition are relatively low. The installation of a teledermatology system increases the number of patients referred for specialist evaluation dramatically, even while the number of in person visits to specialists fell. Although diagnostic agreement between dermatologists and primary care providers was mixed, a marked difference was found in their recommended treatment plans. A number of cases were found in which use of the telemedicine technology system resulted in reversing conditions that had been poorly controlled for a number of years prior to teleconsultation. This work is important as an indicator that referral rates to dermatologists may be inappropriately low in rural areas of the U.S., and that use of teledermatology may improve this trend. PMID- 10384465 TI - Telematics in the neonatal ICU and beyond: improving care, communication and information sharing. AB - In October of 1998, the Beth Israel-Deaconess was awarded one of 19 contracts from the National Library of Medicine (NLM) to develop, implement and test a telemedicine application to support the care of Very Low Birth Weight Infants. This project is the only one to focus on the care of newborns. We believe that this project will provide a new national approach to managing the care of high risk newborns by leveraging evolving communication technology. PMID- 10384467 TI - Telematic evolution of day-hospital in oncology. AB - The telematic evolution of the Day-Hospital can be considered as the virtual hospitalization, implementing the integration between hospital care and home care and predefining an hospital system which "goes to the patient home". In this paper the authors describe tools, characteristics, and advantages of this proposal, as well as the interaction model. Then the medical problem is illustrated: the first model implemented in oncology involves the liver tumors, which are the more frequent malignant neoplasms, and, in particular, the hepatocellular carcinomas (HCC),. The Organization Model, the Information System, the Tools and the Modalities of Telemonitoring for Hepatocellular Carcinoma in telematic day-hospital are still illustrated and a brief conclusion is given. PMID- 10384466 TI - The effect of a neonatal telecardiology system on respiratory therapy in very low birthweight infants. AB - Factors in the U.S. healthcare system have shifted the site of care of many newborns to hospitals where subspecialty services are unavailable. This study examines whether a more rapid turn-around of echocardiogram interpretations and availability of interactive video during neonatal consultations reduces the morbidity of very low birthweight (VLBW) infants. The two groups (n = 21 and n = 28) were similar on the basis of known risk factors. A composite index of respiratory therapy intensivity and duration was used to measure the utilization of respiratory therapies. The index was similar in both groups, 89.6 +/- 12.6 before versus 89.5 +/- 13.0 with telemedicine. These results show little evidence of a reduction in RT utilization. PMID- 10384468 TI - Features for a B-ISDN telemedicine system and its application. AB - Telemedicine technology is emerging as a new way of medical practice. It will provide more cooperative activity between departments and more comfortable access for disabled patients home. We developed two types of telemedicine system: a telediagnosis system and a home care system. Our telemedicine application is aimed to be run on the broadband-integrated services digital network (B-ISDN). The legacy network is also considered. The user interface is designed to help doctors to communicate easily. The key elements of telemedicine systems are user friendly interface, medical multimedia database design highly refined display technique. PMID- 10384470 TI - How can sharing clinical information be made to work? AB - There is a recognised need to share clinical information in order to improve integrity, continuity, safety and speed of delivering patient care. This remains a serious weakness in the conceptualisation of existing health information management systems. The evolution of structured (e.g., HL7) messaging standards has been driven largely from an administrative information viewpoint, as have many of the initiatives driving the development of electronic patient record (EPR) systems. Neither appears to address crucial needs for clinical data exchanges often across wide areas to meet the needs of best quality, cost effective and low risk patient care delivery. The present reality is that clinical messaging is complex, rigid and ineffective, and the business case for its users is not compelling. Administrative and financial arrangements need to be developed which support the more widespread use of clinical data exchanges. This paper underlines the importance of web technology as a key element in the communications strategy and as an adjunct (or alternative) to more structured messaging environments. It also raises some of the fundamental structural problems, which impact the use of messaging in healthcare, and puts forward proposals as to how these may best be addressed and resolved. PMID- 10384469 TI - Functional evaluation of telemedicine with super high definition images and B ISDN. AB - In order to determine whether a super high definition (SHD) image running at a series of 2048 resolution x 2048 line x 60 frame/sec was capable of telemedicine, we established a filing system for medical images and two experiments for transmission of high quality images were performed. All images of various types, produced from one case of ischemic heart disease were digitized and registered into the filing system. Images consisted of plain chest x-ray, electrocardiogram, ultrasound cardiogram, cardiac scintigram, coronary angiogram, left ventriculogram and so on. All images were animated and totaled a number of 243. We prepared a graphic user interface (GUI) for image retrieval based on the medical events and modalities. Twenty one cardiac specialists evaluated quality of the SHD images to be somewhat poor compared to the original pictures but sufficient for making diagnoses, and effective as a tool for teaching and case study purposes. The system capability of simultaneously displaying several animated images was especially deemed effective in grasping comprehension of diagnosis. Efficient input methods and creating capacity of filing all produced images are future issue. Using B-ISDN network, the SHD file was prefetched to the servers at Kyoto University Hospital and BBCC (Bradband ISDN Business chance & Culture Creation) laboratory as an telemedicine experiment. Simultaneous video conference system, the control of image retrieval and pointing function made the teleconference successful in terms of high quality of medical images, quick response time and interactive data exchange. PMID- 10384471 TI - Dental interview system with a native-language interpreting engine. AB - We developed a Dental Interview System with a Native-language Interpreting Engine (DISNIE) on the Internet. DISNIE uses simple natural sentences without dental terminology and interprets these sentences in five languages such as Japanese, English, Korean, Chinese and French, respectively. DISNIE is a good tool not only for patients who are able to read, write and speak only their mother languages but also for the staff at a dental hospital because of its accessibility via the Internet. PMID- 10384472 TI - Collaborative workspace for multimedia medical conferencing. AB - We propose an approach for collaborative workspace management in medical conferencing. A collaborative workspace is a virtual data space shared between medical experts for working out solutions collaboratively while conferencing. Our approach provides medical users with an integrated view of various kinds of multimedia patient data and a unified control over the workspace. For data navigation and conferencing, a tree-like navigation tool, which we named the patient record tree, is provided. And we classify patient data, which is the object of medical collaborative works, into six basic types, and provide a view template for displaying each of these types. PMID- 10384473 TI - Long-term evaluation of the CancerNet WWW service. AB - To improve the quality of healthcare, we offer access to high quality guidelines to physicians as well as information harmonized with these guidelines to concerned patients via internet. We launched this service in 1994. Since then, we have been offering user-friendly access to the NCI's CancerNet via WWW (http://www.meb.uni-bonn.de/). Among other information, CancerNet by the National Cancer Institute contains up-to-date summaries on the prognosis, staging, and treatment of more than 80 major tumor types. To obtain information about how users navigate through this service, all users' activities are logged by using cookies for tracking them without injuring privacy. To get additional information about our users and their interests regarding our service, we performed user surveys in 1996 and 1997. The analysis of 538 valid answers in 1996 and 1001 in 1997 show that the attempt to bring high quality information and guidelines to physicians and patients was successful. About 95% of our users rated our service "excellent" or "good". PMID- 10384474 TI - Disseminating multimedia protocols over Internet for emergency and catastrophe management. AB - Over the last years we have developed various computing methods to assist specialized personnel on various aspects of catastrophe and emergency management. New models can address tasks such as patient triage; stabilization, resource coordination and hospital alertness and techniques based on information technologies. In this paper we present various tools (written on Java and C+2) that we created to store, represent, and disseminate practice guidelines and protocols over the World Wide Web. Guidelines and protocols are stored using a standard database program (e.g., Microsoft Access), and represented in a flowchart format linked to multimedia information such as text, pictures, sound, video or external sources of data. Using our JAVA tool, protocols can be disseminated over the Web and viewed with any browser with JAVA compliance. We have implemented 15 emergency protocols that we developed in collaboration with specialized military personnel from the Ministry of Defense, Spain. Users can access remotely those electronic protocols comparing their procedures and methods. Our goal is to enhance agreement and consensus among remote medical centers regarding emergency and catastrophe management, establishing discussions over the network. Our tools have also a potential for training medical and paramedical personnel for emergency situations. PMID- 10384475 TI - Medical counseling through computer communication networks. AB - Medical counseling on the computer communication network will be used increasingly with wide-ranging concern of people about health and strengthening the role of medical care as social service. This study was designed to explore main problems/issues of network counseling and to suggest future prosperity of medical counseling. We analyzed questions and answers in the Association of Family Medicine in the KETEL since 1991, Medical counseling corner which is in the UNITEL since 1996, and Online counseling in VIRTUAL HOSPITAL since 1996, and classified the reason for encounter, systematic classification of counseling by ICPC, and answer content. Inquiry about symptoms was the most common reason for encounter, followed by treatment and prevention strategies, detailed information about diseases, and inquiry about laboratory tests. Systematic classification by ICPC showed common problems of skin, digestive, musculoskeletal and general & unspecified, which was a different look compared with common problems in primary care. The answer content presented a different look among three groups. Common answers were mainly to offer medical information, to recommend visiting a doctor, and to provide self-care remedies. Network counselling will supplement the incompleteness of medical interview with doctors, and make it easy to communicate various pieces of information between patients and doctors. PMID- 10384476 TI - IMGT, the international ImMunoGeneTics database: a new design for immunogenetics data access. AB - IMGT, the international ImMunoGeneTics database is an integrated database specializing in Immunoglobulins (Ig), T-cell receptors (TcR) and MHC molecules of all vertebrate species, created by Marie-Paule Lefranc, University of Montpellier, CNRS, Montpellier, France (Nucleic Acids Research, Database issue, Vol 26, January 1998). IMGT includes three databases: LIGM-DB (for Ig and TcR), MHC/HLA-DB and IMGT/PRIMER-DB (an Ig, TcR and MHC-related primer database), the last two in development. IMGT comprises expertly annotated sequences and alignment tables. LIGM-DB contains more than 24.000 Immunoglobulin and T cell Receptor sequences from 81 different species. MHC/HLA-DB contains class I and class II Human Leucocyte Antigen alignment tables. An IMGT tool, DNAPLOT, developed for Ig, TcR and MHC sequence analysis, is also available. IMGT goals are to establish a common data access to all immunogenetics data, including nucleotide and protein sequences, oligonucleotide primers, gene maps and other genetic data of Ig, TcR and MHC molecules, from all species, and to provide a graphical user friendly data access. IMGT has important implications in medical research (repertoire in autoimmune diseases, AIDS, leukemias, lymphomas), therapeutical approaches (antibody engineering), genome diversity and genome evolution studies. In this paper, we describe our approach for the data modelisation, the automation of the annotation procedure and control of data quality in LIGM-DB database. IMGT is freely available on the CNUSC WWW server at Montpellier: http://imgt.cnusc.fr: 8104 (contact: Denys.Chaume@cnusc.fr) and on the EBI servers: http://www.ebi.ac.uk/imgt (contact: malik@ebi.ac.uk) and ftp.ebi.ac.uk/pub/databases/imgt. LIGM-DB users are encouraged to report errors or suggestions to giudi@ligm.crbm.cnrs-mop.fr. IMGT initiator and coordinator: Marie-Paule Lefranc, lefranc@ligm.crbm.cnrs-mop.fr. (fax: +33(0)467040231). PMID- 10384477 TI - Integration of bioInformatics tools at the National University of Singapore (NUS). AB - In the past decade "Big Science" such as the Genome Project has generated an enormous amount of data in the life sciences. Concurrently, the synergy of this project with existing research has quickened the pace of biological discovery. But the major drawback that is beginning to be felt worldwide is the primitive level of organisation in the data accumulated. Without a proper framework or knowledge scaffold to hang and interconnect the various bits of data and information, the national knowledge-to-data ratio is declining rapidly. We are trying to serve a solution to this enigma by providing a World Wide Web (WWW) interface to Biosoftware and at the same time have come up with a database integration tool that can query heterogeneous, geographically scattered and disparate databases simultaneously. In this report we will talk about BioInformatics in general with specific reference to BioInformatics Centre (BIC) at the National University of Singapore. PMID- 10384478 TI - A integrative molecular information system. AB - Methods of biotechnology allow the isolation, sequencing, and synthesis of molecular structures. Molecular database systems are available, which allow the worldwide data access. Methods and concepts of bioinformatics are important for the analysis of these molecular data, which represent complex regulatory networks. In this paper we present the architecture of our molecular information system. PMID- 10384479 TI - Structural-functional bioinformatics: knowledge-based NMR interpretation. AB - This paper describes a knowledge-based approach to a problem of structural functional bioinformatics, specifically the determination of protein structure through the automated analysis of NMR data. Highly successful results in carrying out sequence-specific assignments of residues from multidimensional NMR datasets has led us to automation of NOE dataset interpretation and a design for integrating these results with other protein structure and function analysis programs. PMID- 10384480 TI - The application of model-based complexity inference method to molecular evolution analysis. AB - In this study, a new method based on the concept of complexity in inductive inference is proposed for reconstructing molecular phylogenetic tree. This method describes the complexity of molecular phylogenetic tree by three terms, which are related to tree topology, the branch lengths and fitness between the model and data measured by likelihood function. The computer simulation is used to investigate the efficiency of this method. The results suggest that this method is superior to the traditional methods because it avoids excess-complexity in the tree model estimation available from DNA sequence. PMID- 10384481 TI - Modeling and simulation in molecular pharmacology. AB - A stochastic simulation program of drug-receptor interaction is presented. The user can select a set of conditions concerning the processes of response release (type of cellular response, drug distribution and metabolism etc.) and the program plots a family of dose-response curves. A comparison between experimental and simulated dose-response curves shows the validity of the model can be used either for testing hypotheses concerning drug-receptor interaction, for experimental design or for estimation of some specific parameters. The program can also be used for educational purposes. PMID- 10384483 TI - Modelling human depth perception in binocular vision: obtaining the horizontal disparity map. AB - The paper presents a computer application developed as a tool for implementing, developing, and testing computational models for stereopsis. Two models for solving the correspondence problem and computing the stereo disparity map have been implemented. One of them is biologically inspired (it models the behaviour of simple and complex cells from the striate cortex) and the paper details the results obtained on random-dot stereograms and on pairs of real images. PMID- 10384482 TI - Simulation of the initial concentration-time course after intravenous application of the drug. AB - In this paper we present a widely applicable computational method for the description of the initial concentration-time-course after intravenous injection of a substance. The intravascular concentration-time course, r, is described as r = c0 + g x r, where the asterisk denotes the convolution operation, c0 is the concentration-time course during the first passage of the substance and g is the transport function of the body. If the body transport function is known, then the concentration-time course of a substance can be predicted. The site of interest can be chosen arbitrarily, i.e. the concentration-time course in the arterial circulation supplying any organ can be described. This might be of special interest for the optimal design of intravenous injections of contrast media, where initial concentrations at the region of interest determine the success of the diagnostic procedure. PMID- 10384485 TI - Population size and quality in genetics-based rule learning from medical data. AB - Population size and quality are parameters which control the performance of genetic algorithms. We researched these parameters in a genetic-based machine learning system Galactica which was used to discover the differential diagnostic rules for female urinary incontinence from case data. The performance of the system was measured with on-line and off-line criteria. Surprisingly, randomly generated small populations (30 and 70 rules) did not promote the best on-line performance as earlier results suggested. Probable explanation is the lack of diversity in initial populations. The seeding of population with positive learning examples was used to obtain more divergent populations. As expected, the seeding increased the on-line performance of small populations. The results are mainly in accord with the earlier results indicating that large randomly generated populations (150 rules) lead to the better off-line performance. Again, the seeding of small populations was successful producing even the better off line performance than a large population. In conclusion, the seeding allowed the small populations to converge to good rules in relatively short period of time. PMID- 10384484 TI - Cepstral distance measures of hormone concentration time series. AB - In this paper we present a class of time series distance measures based on the difference of their cepstral transformations. We emphasise the convenience of the proposed distance measure in the cases when the time series can be treated as output of a linear system driven with a quasi-periodic stochastic signals. In order to illustrate the cepstral time series distance measure we applied them in cluster and multidimensional scaling analysis of daily hormonal secretion fluctuation series taken from a group of patients before and after surgery. PMID- 10384486 TI - Preparing for the third millennium: the views of life informatics. AB - The chief aspects of this paper are the condition of the birth of life informatics and its tasks, basic concepts, principles, and structure. There are three phases of combining informatics with medicine: product, technological, and theoretic application of which the goals are respectively the informatization of numerical and word processing, data of medical treatment, and the knowledge of medicine. While reached the third phase we have dealt with two types of biological information, physical and nonphysical, i.e., body information (i.e., the information about body's components and structure), and life information (i.e., the information about life codes and life programs). Life informatics is a main branch of bioinformatics. It is a new member of the medical informatics family, and as such is younger than health informatics, nursing informatics, and dental informatics. It's task is to assist biologists and medical doctors to recognize and interfere the human life information procedure just as they are doing well with human body's matter and energy system. Its basic concepts are life information, life information medicine, and life information therapy. Its most important principles are information materialism, general informatics, and information determinism. Its main branches are biomolecule, cellular, organic, individual, and social informatics. In the third millennium, the life informatics will be a leading discipline in biology, medicine and informatics, which will gradually influence modern philosophy and other humanities. PMID- 10384487 TI - Self-documenting structured reports using open information standards. AB - Structured reporting systems use standardized data elements and predetermined data-entry formats to record observations. This article describes a system for structured data entry and reporting that generates reports encoded in the Standard Generalized Markup Language (SGML), an open, internationally accepted standard for document interchange. The structured report is self-documenting: it includes a definition of its allowable data field and values encoded as a report specific SGML document type definition (DTD). By linking its reporting concepts with those of external vocabularies such as the UMLS Metathesaurus, this system can create open, universally comprehensible structured reports. PMID- 10384488 TI - Sending specialist reports to GPs using EDI. AB - This paper describes a project in which specialist reports are sent to general practitioners using EDI. The reports were protocolled to better meet the wishes of the GPs. This also made it possible to achieve a further structurization of the existing EDI-message, using free-text qualifiers. Future developments of specialist-GP communication are discussed, specifically regarding the integration of their electronic medical records. It is concluded that EDI can and will play an important role in this. PMID- 10384489 TI - Shared care for diabetes: supporting communication between primary and secondary care. AB - As health care becomes more complex, interest in the benefits of coordination of care has increased. Especially patients that are being treated jointly by more than one physician (shared care), are vulnerable to adverse effects resulting from inadequate coordination and communication. We describe a study in which care providers support shared care by using computer-based patient records for data storage, and structured electronic data interchange (EDI) as a means of communication. The study showed that the electronic communication network for exchanging consultation outcomes significantly increased frequency of communication and the availability of data to the general practitioner on diagnostic procedures performed in the hospital, thus providing more complete information about the care that patients are receiving. PMID- 10384490 TI - Medical Internet exchange project in Japan. AB - The Internet has been widely used by medical institutes and hospitals around the world, however; its use for telemedicine is still low. The main reason for this is the availability of bandwidth and poor security through the net. Meanwhile, we have established and have been operating 'Cancer Information Network' among 11 Cancer Centers in Japan, mainly for Multipoint TV Conference using HDTV image. There are also similar projects among 9 cardiovascular centers in Japan. By March, all 240 national hospitals will have been connected by an IP network using an ATM backbone. The above network projects are operated independently, and have an 'Intranet' characteristics within them. There are also many hospitals and clinics connected to the Internet by commercial internet providers. To make a secure and efficient network between these medical networks and medical sites, we started the Medical Internet eXchange project (MDX project) constructing a Medical Network Operation Center to create a link between them. To provide the administrative policy of this project, we established the Medical Internet eXchange Association. We are planning to expand this project to Asian-Pacific countries using the Asian-Pacific Advanced Network (APAN), and also expand it to worldwide connections in the future. For this purpose, we are currently asking other countries to form a structure similar to MDX-Japan. The concept, hardware system, software system, firewall configuration, and routing policy will be also discussed. PMID- 10384492 TI - Achieving standardization of health information in Canada by the year 2000. AB - As health reforms gain momentum in Canada, a critical need has been identified for the development of standardized health information across the continuum of health services. Although vast amounts of data are collected, deficiencies and gaps in current information systems seriously limit its effective use. As provinces and territories move toward more integrated health services systems, the necessity for a comprehensive and systematic analysis of health information requirements, and the development of requisite standards has never been greater. To begin addressing this need, the Canadian Institute for Health Information (CIHI) established a number of interrelated projects in 1995. This paper describes the goals of these projects, as well as the approach used to assess information needs across health services and to develop required standards. PMID- 10384491 TI - University medical information network--past, present, and future. AB - The University Medical Information Network (UMIN), established in 1989, is a network service organization for national university hospitals in Japan. It has provided various medical information services to medical professionals, including database, electronic mail, and news services. Although its initial network was constructed as a closed network using N1 protocol, it now adopts TCP/IP protocol and is open to other medical professionals via the Internet. The next UMIN network system is planned to be constructed as a secure virtual closed network on the Internet, using cipher technology, and to provide secure information services to national university hospitals via the closed network, and to other medical professional via the Internet. User friendly interface and flexible system development were made possible by adopting TCP/IP, and the number of users dramatically increased accordingly. However, the database, software design, and human organizations developed in the N1 era have now proven to be of great value, and contribute to todayis flourishing state of the UMIN. PMID- 10384493 TI - Standardization of the nationwide health examination in Korea. AB - The Korea Medical Insurance Corporation has held the periodic Health Examination for the public servants and teachers from 1977 as a nationwide health preventive task. But the Health Examination result is not computerized rather than paperwork, so the use of the Health Examination is limited and the results of Health Examination can not be stored or interchangeable between hospitals or health examination centers in this system. So we planed the Standardization of the Nationwide Health Examination project and developed the Health Examination computer program in 1996. The object of standardized program is to contribute to cost-effective analysis of each item or other studies about the factors influences development of diseases. And now, an Exhibition work for the standardization is being held with the Health Examination computer program. At the ending of the Exhibition work, we will evaluation and revision the computerized program for standardization process. PMID- 10384495 TI - Proposal of a new Internet standard for DICOM: DICOM-QR URL. AB - This paper proposes a new Internet standard that is combination of two standards in different domain, Internet and medical informatics. The both standards are described briefly in this paper. We describe how to combine them into a Internet standard. With a new standard, there are several advantages for medical information systems. The standard should be established by the following way of the Internet standards. PMID- 10384494 TI - Patient information exchange guideline MERIT-9 using medical markup language MML. AB - To realize clinical data exchange between healthcare providers, there must be many standards in many layers. Terms and codes should be standardized, syntax to wrap the data must be mutually parsable, then transfer protocol or exchange media should be agreed. Among many standards for the syntax, HL7 and DICOM are most successful. However, everything could not be handled by HL7 solely. DICOM is good for radiology images, but, other clinical images are already handled by other "lighter" data formats like JPEG, TIFF. So, it is not realistic to use only one standard for every area of clinical information. For description of medical records, especially for narrative information, we created SGML DTD for medical information, called MML (Medical Markup Language). It is already implemented in more than 10 healthcare providers in Japan. As it is a hierarchical description of information, it is easily used as a basis of object request brokering. It is again not realistic to use MML solely for clinical information in various level of detail. Therefore, we proposed a guide-line for use of available medical standards to facilitate clinical information exchange between healthcare providers. It is called MERIT-9 (MEdical Records, Images, Texts,--Information eXchange). A typical use is HL7 files, DICOM files, referred from an MML file in a patient record, as external entities. Both MML and MERIT-9 are research projects of Japanese Ministry of Health and Welfare, and the purpose is to facilitate clinical data exchanges. They are becoming to be used in technical specifications for new hospital information systems in Japan. PMID- 10384496 TI - Development of medical decision support system for leukemia management. AB - A leukemia management system was developed in this study which is comprised of four modules: registry, diagnosis, prognosis of treatment, and CAI. The emphasis was on patient management as a whole. Three knowledge models were developed to predict accurate diagnosis and prognosis of treatment: case-based reasoning, neural network, and discriminant analysis, Of these, discriminant analysis model produced the most accurate diagnosis, whereas neural network produced the most accurate prognosis of treatment. PMID- 10384498 TI - The future of knowledge-based components in the electronic health record. AB - Traditional monolithic healthcare information systems (HIS) no longer meet the requirements of today's distributed enterprises and the rapidly changing healthcare environment. The ability of applications to communicate, interpret, and act intelligently upon complex healthcare information has assumed paramount importance. The future lies in the development of flexible component-based architectures that can operate seamlessly within the workflow of a healthcare environment. A key design goal is "graceful degradation," i.e., providing the best decision support possible within the context of available patient data. The First DataBank Drug Toolkit is used as a case study. Several technical challenges associated with building truly plug and play components are discussed. PMID- 10384497 TI - Hospital management decision support: a balanced scorecard approach. AB - Hospital management teams receive voluminous data from a wide variety of sources, but are unable to distill the essential data they require to make good decisions. We have used a methodology, which helps teams define and use important management data coupled with an information system that makes this data accessible. Results of our evaluation indicate that the process of developing a Balanced Scorecard indicator system helps management teams to define meaningful strategic objectives and measurable performance indicators. The framework combined with the information acts as an integrating force, providing a shared understanding of the unit's goals. We conclude that a customized decision support system, which integrates multiple measures in a balanced Scorecard framework, is a powerful tool for enabling complex decision making by a management team. PMID- 10384499 TI - A data dictionary approach to multilingual documentation and decision support for the diagnosis of acute abdominal pain. (COPERNICUS 555, an European concerted action). AB - This paper describes the design and development of a multilingual documentation and decision support system for the diagnosis of acute abdominal pain. The work was performed within a multi-national COPERNICUS European concerted action dealing with information technology for quality assurance in acute abdominal pain in Europe (EURO-AAP, 555). The software engineering was based on object-oriented analysis design and programming. The program cover three modules: a data dictionary, a documentation program and a knowledge based system. National versions of the software were provided and introduced into 16 centers from Central and Eastern Europe. A prospective data collection was performed in which 4020 patients were recruited. The software design has been proven to be very efficient and useful for the development of multilingual software. PMID- 10384500 TI - Decision support for medication use in an inpatient physician order entry application and a pharmacy application. AB - Studies have shown that adverse drug events are common, expensive, and due to causes that can be remedied by information technologies. At our institution we have developed a physician order entry application and a pharmacy application designed to decrease the risk of such adverse drug events. In this paper, we describe the applications, with attention to the clinical decision support features present in each. We also describe the manner in which the two applications interact. PMID- 10384501 TI - Simple models for estimating dementia severity using machine learning. AB - Estimating dementia severity using the Clinical Dementia Rating (CDR) Scale is a two-stage process that currently is costly and impractical in community settings, and at best has an interrater reliability of 80%. Because staging of dementia severity is economically and clinically important, we used Machine Learning (ML) algorithms with an Electronic Medical Record (EMR) to identify simpler models for estimating total CDR scores. Compared to a gold standard, which required 34 attributes to derive total CDR scores, ML algorithms identified models with as few as seven attributes. The classification accuracy varied with the algorithm used with naive Bayes giving the highest. (76%) The mildly demented severity class was the only one with significantly reduced accuracy (59%). If one groups the severity classes into normal, very mild-to-mildly demented, and moderate-to severely demented, then classification accuracies are clinically acceptable (85%). These simple models can be used in community settings where it is currently not possible to estimate dementia severity due to time and cost constraints. PMID- 10384502 TI - Induction of diagnostic test strategies with multi-level information measures. AB - This paper presents a method for inducing clinical diagnostic test protocols or strategies from data. We represent testing strategies as a strategy tree. To support induction of strategy tree, we define three information measures: K-level information, K-level information gain, K-level gain ratio, and K-level cost index, for test selection during strategy building. These measures generalize Quinlan's information measures used in decision tree induction. We present theoretical and experimental results to show that the K-level cost index can be used to induce strategy trees in a practical domain. PMID- 10384503 TI - Modelling medical decisions in DynaMoL: a new general framework of dynamic decision analysis. AB - Dynamic decision analysis concerns decision problems in which both time and uncertainty are explicitly considered. We present a new dynamic decision analysis framework, called DynamoL, that supports graphical presentation of the decision factors in multiple perspectives. To alleviate the difficulty in assessing conditional probabilities over time in dynamic decision models, DynaMoL incorporates a Bayesian learning system to automatically learn the probabilistic parameters from large medical databases. We describe the DynaMoL modeling and learning architecture through a medical decision problem on the optimal follow-up schedule for patients after curative colorectal cancer surgery. We also show that the modeling experience and results indicate practical promise for the framework. PMID- 10384504 TI - Architecture of a neural network client/server system for decision support in clinical information systems. AB - Neural networks have demonstrated their research potential in bio-medical sciences. The literature is filled with experiments that evidence the value of neural networks for the solution of diagnostic and predictive problems. The next step is to direct research efforts toward implementation of these neural networks into the clinical information environments where their diagnostic and predictive capabilities can be truly utilized. To accomplish this goal, it is necessary to create a new generation of neural network tools that ease the transition of experimental results to a production state. This paper describes the architecture of such a system. PMID- 10384505 TI - Using classification tree and logistic regression methods to diagnose myocardial infarction. AB - Early and accurate diagnosis of myocardial infarction (MI) in patients who present to the Emergency Room (ER) complaining of chest pain is an important problem in emergency medicine. A number of decision aids have been developed to assist with this problem but have not achieved general use. Machine learning techniques, including classification tree and logistic regression (LR) methods, have the potential to create simple but accurate decision aids. Both a classification tree (FT Tree) and an LR model (FT LR) have been developed to predict the probability that a patient with chest pain is having an MI based solely upon data available at time of presentation to the ER. Training data came from a data set collected in Edinburgh, Scotland. Each model was then tested on a separate Edinburgh data set, as well as on a data set from a different hospital in Sheffield, England. Previously published models, the Goldman classification tree[1] and Kennedy LR equation[2], were evaluated on the same test data sets. On the Edinburgh test set, results showed that the FT Tree, FT LR, and Kennedy LR performed equally well, with ROC curve areas of 94.04%, 94.28%, and 94.30%, respectively, while the Goldman Tree's performance was significantly poorer, with an area of 84.03%. The difference in ROC areas between the first three models and the Goldman model is significant beyond the 0.0001 level. On the Sheffield test set, results showed that the FT Tree, FT LR, and Kennedy LR ROC areas were not significantly different (p > = 0.17), while the FT Tree again outperformed the Goldman Tree (p = 0.006). Unlike previous work[3], this study indicates that classification trees, which have certain advantages over LR models, may perform as well as LR models in the diagnosis of patients with MI. PMID- 10384506 TI - Implementation of a fuzzy prototype-based machine learning method to predict myocardial infarction from coronary angiography. AB - Formal knowledge on the predictive value of morphological angiographic factors is lacking to estimate the risk of myocardial infarction. This article presents a computer system for predicting the incidence of myocardial infarction from angiographic morphological descriptions of coronary lesions. The system includes two phases. The learning phase consists in extracting from a large database of described stenoses two classes represented by one or several fuzzy prototypes. One class corresponds to stenoses leading to infarction and the other to stenoses not leading to that event. The evaluation phase consists in classifying a stenosis according to its morphological characteristics in one of these two classes. The learning method is based on a fuzzy supervised Machine Learning algorithm that combines some aspects of the K-nearest neighbours clustering approach with a defined measure of similarity, and a prototype induction function from the most similar stenoses, taking into account their degree of typicality. The current results of the evaluation phase to correctly predicted X% stenoses for their risk of myocardial infarction. This article emphasizes the feasibility of the approach, however, the learning phase relies on some heuristics that should be validated to get a formal evaluation of the system. PMID- 10384507 TI - Consistency management in multiple perspective medical decision analysis. AB - Multiple perspective reasoning is often involved in real-world decision analysis. Different perspectives may be suitable for different stages of the decision modeling process. Multiple perspective modeling calls for consistency management which ensures that the different perspectives reflect the same information. This paper summarizes the graphical perspectives currently supported in DynaMoL, a new framework for dynamic decision analysis. We introduce a new perspective, the tree view, into the framework. We present the main ideas involved in consistency management in the framework. We also discuss the critical issues involved in multiple perspective modeling of a simplified case study in medicine. PMID- 10384508 TI - The nurse scheduling problem: a combinatorial problem, solved by the combination of constraint programming and real users heuristics. AB - Nurse scheduling consists of assigning shifts and rest to nurses for each day, on a time schedule, taking in account legal and collective constraints, and individual wishes. The Nurse Scheduling Problem (NSP) is a highly difficult and complicated problem that has already be explored using several operational research methods. Nevertheless, those methods are not flexible enough to match the individual requests from nursing staff and they don't allow for effective management of unforeseen absences. This paper explores the use of a Constraint Programming (CP) to build Gymnaste, which is since June 1997 a available package, after five years of a slow research and development process. We describe the way the problem is modeled, the constraints typology, and the interface. In fact, the NSP is also an ill defined problem, making the man computer interface critical to let the user chose the best heuristics. Gymnaste is currently beta-tested on several pilot sites. Schedules have been generated very quickly, typically in less than a minute for 20-30 persons over 4 weeks. An evaluation process is running, which preliminary results also show some interesting sociological and organizational aspects. PMID- 10384509 TI - Knowledge discovery and case based reasoning in health promotion: development of a help-desk for prevention of occupational injuries. AB - This paper presents the concepts, ideas and techniques behind Case Based Reasoning (CBR) in relation to knowledge extraction techniques for health promotion. The ultimate goal is to develop a help-desk service for advice about preventive measures to be taken concerning concrete occupational injury hazards. CBR has been suggested to be a complimentary method to knowledge extraction in order to take direct advantages of large databases for building decision support systems. In this work a database on work injuries is being used to develop a CBR application using a CBR shell-called Recall. PMID- 10384510 TI - Relevant information for decision support systems: application in cardiology. AB - In the paper we show information theory tools for extracting relevant information for decision support systems from medical databases. Each proposed algorithm for selecting a set of relevant features has a specific score function defined by means of information-theoretical characteristics. Then algorithms are classified according to the primary criterion, that can lead to influence-preferring algorithms or weight-preferring algorithms. Other type of classification can be based on the way of selecting of features as forward, backward or combined algorithms. The software package called CORE (COnstitution and REduction) that supports the process of selection of features relevant for a decision making problem is described. Application on data about 1417 middle age men collected in the twenty years lasting interventional study of cardiovascular risk factors in middle aged men and for decision support in primary care are shown. However, the methodology presented is applicable for any decision making problem where extracting relevant information from data is required. PMID- 10384511 TI - Presenting treatment protocols with Web technology. AB - This paper describes a Web-based version of a protocol information system (ProtoVIEW) with which a wide range of diagnostic or therapeutic protocols can be retrieved and viewed. The Web-based version contains all functionalities of the non web-based version plus several new functionalities. The web version contains an X-ray viewer and a great deal of interactivity such as validation of electronic patient data forms. The most important additional function is the context sensitive protocol support which may lead to improved protocol adherence. Finally, the web-based version can be accessed from any working place since patient data and protocols are stored centrally. PMID- 10384512 TI - A statistical model that takes into account patient heterogeneity in decision making. AB - Statistical evaluation of clinical treatments or preventive medicine has profoundly contributed to decision making in medical fields such as with the acceptance of new treatment methods and health promotion policies. It is crucial in such decision making to find a correct statistical model to treat a surprisingly large variety of patients, or a heterogeneous group of patients, even with the same diagnosis. In diseases such as cancer, cardiovascular disease or diabetes, patients are often followed up to certain endpoints and these data are frequently analyzed by logrank tests or Cox-models to evaluate treatment effects. Although these methods have been widely accepted and extensively studied, we are sometimes faced with problems in applying these methods when the heterogeneity of patients is large and a lot of prognostic factors affecting the endpoints have to be considered. Based on the results of the analyses of survival data from more than 6,000 gastric cancer patients, it is revealed that the stratified logrank test may suffer serious power loss, even though primary prognostic factors are used as stratified factors. A so-called 'piecewise linear Cox regression method' for properly treating the heterogeneity of patients is introduced and extensively studied. This method is shown to be appropriate for patient groups with a high degree of heterogeneity such as the gastric cancer patients. The same method is, in principle, applicable to patients of other diseases, too, using statistical software such as SAS, BMDP and etc. PMID- 10384513 TI - The limitations of decision trees and automatic learning in real world medical decision making. AB - The decision tree approach is one of the most common approaches in automatic learning and decision making. It is popular for its simplicity in constructing, efficient use in decision making and for simple representation, which is easily understood by humans. The automatic learning of decision trees and their use usually show very good results in various "theoretical" environments. The training sets are usually large enough for learning algorithm to construct a hypothesis consistent with the underlying concept. But in real life it is often impossible to find the desired number of training objects for various reasons. The lack of possibilities to measure attribute values, high cost and complexity of such measurements, unavailability of all attributes at the same time are the typical representatives. There are different ways to deal with some of these problems, but in a delicate field of medical decision making, we cannot allow ourselves to make any inaccurate decisions. We have measured the values of 24 attributes before and after the 82 operations of children in age between 2 and 10 years. The aim was to find the dependencies between attribute values and a child's predisposition to acidemia--the decrease of blood's pH. Our main interest was in discovering predisposition to two forms of acidosis, the metabolic acidosis and the respiratory acidosis, which can both have serious effects on child's health. We decided to construct different decision trees from a set of training objects, which was complete (there were no missing attribute values), but on the other hand not large enough to avoid the effect of overfitting. A common approach to evaluation of a decision tree is the use of a test set. In our case we decided that instead of using a test set, we ask medical experts to take a closer look at the generated trees. They examined and evaluated the decision trees branch by branch. Their comments on the generated trees can be found in this paper. The comments show, that trees generated from available training set mainly have surprisingly good branches, but on the other hand some are very "stupid" and no medical explanation could be found. Thereafter we can conclude, that the decision tree concept and automatic learning can be successfully used in real world situations, constrained with the real world limitations, but they should be used only with the guidelines of appropriate medical experts. PMID- 10384514 TI - Microcomputer decision support system for intensive care. AB - This article describes a computer system to support decisions in Intensive Medicine developed for a multipurpose Intensive Care Unit (13 beds and about 700 yearly patients). The system is used in a microcomputer (486DX4). Works done in this area are mentioned; and the different programs which integrate the system, the advantages of its use and the results accomplished are described. Some of the most important results are the quality increment of the medical care given, physicians' possibilities of dedicating more time to their patients, cost reductions, and improvements in the development of teaching and in the conditions to carry out research work in the Intensive Care Unit (ICU). An analysis is made of the system requirements that guarantee its integration to the Unit work, and the easy and safe use of it by the physicians and nurses. PMID- 10384515 TI - Diagnosing breast cancer from FNAs: variable relevance in neural network and logistic regression models. AB - We compared the selection of variables for building a classification model for the diagnosis of breast cancer using neural networks and logistic regression. A set of 460 cases was used to build neural network and logistic regression models that classify cell samples obtained by fine-needle aspiration (FNA) as malignant or benign, depending on nine pathology features. Variables selected by a step down logistic regression model were compared to those selected by a measure of relevance derived from neural network weights. Since both types of models resulted in similar predictive accuracy, we expected approximately the same variables to be selected. The variables with the highest relevance values for the neural network models corresponded to those of high significance in univariate logistic regression models, but were not the ones selected in the step down procedure of multivariate models. Variable relevance based on weights for neural network models does not seem to be a consistent index of the importance of that variable for multivariate models such as logistic regression. PMID- 10384516 TI - Modelling health care processes with SHARE. AB - This paper describes the "SHARE" workframe, designed to provide a comprehensive environment for modeling and simulating health care processes. The objects defined within SHARE are Actors, subdivided in Clients and Resources, and Elementary Operations. Graphical tools allow to build processes from these objects, and to specify their relationships. Various strategies based on either clinical or managerial changes may be investigated. Summary information on the utilization of all actors, on waiting times and goodness of execution may be displayed after a simulation. Better description of processes, and their study a priori will improve reliability, quality of care and satisfaction of patients. PMID- 10384517 TI - Case based diagnosis in histopathology of breast tumours. AB - Relevant knowledge and decision making process in histopathology is mostly included in typical pathological cases encountered by the expert. In this article we address the issue of exploiting this knowledge in the diagnosis process. We present the first steps of a Case-Based-Reasoning (CBR) system that uses the previously resolved pathological cases in order to facilitate decision making and diagnosis formulation of a new case. The work has been performed in two phases. Firstly, an object-oriented model of the domain was developed and 35 pathological cases of breast tumours were represented within this model. Secondly, the functional architecture of the CBR system was designed and the main procedure, the selection of similar cases, was achieved. The selection procedure is based on an original similarity measure that takes into account both semantic and structural resemblances and differences between the cases. A first evaluation of the system was performed on several cases of the data base. The interest of the CBR approach in situations where heuristic rules cannot be clearly defined is discussed. PMID- 10384518 TI - Focusing on resistance development in a case based teleconsultation system for antibiotics therapy advice. AB - In this paper, we describe an approach to support physicians when they select a calculated antibiotic therapy for intensive care patients who have developed an infection as an additional complication. As advice is needed quickly and the pathogen is not yet known, we use an expected pathogen spectrum based on medical background knowledge and known resistances, which both will be adapted to the results of the laboratory. Case-Based Reasoning retrieval methods provide the advice for similar previous patients. Their solutions are adapted to be applicable to the new medical situation of the current patient. Furthermore, we present the recent resistance developments of the antibiotics to the physician. PMID- 10384519 TI - Prognoses of multiparametric medical time courses applied to kidney function assessments. AB - In this paper, we describe an approach to utilize Case-Based Reasoning methods for trend prognoses for the monitoring of the kidney function in an Intensive Care Unit (ICU) setting. Since using conventional methods for reasoning over time does not fit for course predictions with poor medical knowledge of typical course patterns, we have developed abstraction methods suitable for integration into our Case-Based Reasoning system ICONS. These methods combine medical experience with prognoses of multiparametric courses. On the ICU, the monitoring system NIMON provides a daily report based on current measured and calculated kidney function parameters. We subsequently generate course-characteristic trend descriptions of the renal function over the course of time. Using Case-Based Reasoning retrieval methods, we search in the case base for courses similar to the current trend descriptions. Finally, we present the current course together with similar courses as comparisons and as possible prognoses to the user. We applied Case Based Reasoning methods in a domain which seemed reserved for statistical methods and conventional temporal reasoning. PMID- 10384520 TI - Modelling computerised clinical pathways. AB - Since the mid 1980s, paper clinical pathways have been used in defining the road map of patient care. They have been used with varying degree of success for providing more cost-effective healthcare and helped to establish quality improvement models for healthcare delivery. Many attempts have been made to produce electronic versions of the paper clinical pathways in order to maximise benefits of the paper based systems. However, all paper systems are designed based on linear sequential model with little decision support capability. Current electronic versions of the paper systems produce only minimal improvements on the functionality of their paper counterparts. A state-transition information model (STIM) grounded in the Object Oriented system design paradigm is used to reconceptualise a computerised clinical pathways design. A computerised clinical pathways prototype is currently being developed based on this STIM model. The prototype will demonstrate improved functionality: better information management and decision support capabilities. PMID- 10384521 TI - Automatic extraction of linguistic knowledge from an international classification. AB - Automatic extraction of knowledge from large corpus of texts is an essential step toward linguistic knowledge acquisition in the medical domain. The current situation shows a lack of computer-readable large medical lexicons, with a partial exception for the English language. Moreover, multilingual lexicons with versatility for multiple languages applications are far from reach as long as only manual extraction is considered. Computer-assisted linguistic knowledge acquisition is a must. A multilingual lexicon differs from a monolingual one by the necessity to bridge the words in different languages. A kind of interlingua has to be built under the form of concepts to which the specific entries are attached. In the present approach, the authors have developed an intelligent rule based tool in order to focus on a multilingual source of medical knowledge, like the International Classification of Disease (ICD) which contains a vocabulary of some 20,000 words, translated in numerous languages. PMID- 10384522 TI - Acquisition of lexical resources from SNOMED for medical language processing. AB - Medical language processing depends on large-coverage, fine-grained specialized lexicons. The vast majority of existing electronic lexicons concern the English language; for other languages such as French, resources are scarce. In contrast, large medical thesauri exist in numerous languages, including French. Our goal was to study what kind of linguistic information could be extracted from thesauri into a lexicon, in which places human intervention is necessary, and what kind of issues arise in this process. We designed in this purpose a method to build a semantic lexicon from a subset of the SNOMED axes in their French translation. PMID- 10384523 TI - Natural language generation of surgical procedures. AB - The GALEN-IN-USE project has developed a compositional scheme for the conceptual representation of surgical operative procedure rubrics. The complex representations which result are translated back to surface language by a tool for multilingual natural language generation. This generator can be adapted to the specific characteristics of the scheme by introducing particular definitions of concepts and relationships. We discuss how the generator uses such definitions to bridge between the modelling 'style' of the GALEN scheme and natural language. PMID- 10384524 TI - A collaborative approach to building a terminology for medical procedures using a Web-based application: from specifications to daily use. AB - The MAOUSSC (Model for Assistance in the Orientation of a User within Coding Systems) Web server supports a collaborative work on the description of medical procedures. The specifications for the MAOUSSC application are conceptual modeling, definition of semantically fully described procedures, re-use of an existing vocabulary, the UMLS, and sharability. This paper reports on some difficulties in applying those principles in a networked building and updating of the terminology. The users are physicians who have to represent procedure terms in the MAOUSSC formalism. They must apply the constraints of the underlying model, and re-use the representation of the UMLS knowledge base. In our experience, we found that the implementation of syntactic and semantic constraints was not sufficient. Guidelines for pragmatical aspects in representation are required to make a collaborative approach in terminology building more operational. PMID- 10384525 TI - Integrating sources for a clinical reference terminology: experience linking SNOMED to LOINC and drug vocabularies. AB - Achieving the promise of higher quality, lower cost and more available health care through electronic medical records requires the support of a comprehensive clinical reference terminology. In a previous paper we described SNOMED RT (reference terminology), and the data structures and logic syntax that support the transformation of the SNOMED III nomenclature into the SNOMED RT reference terminology. In this paper, we describe an approach to linking SNOMED RT to existing nomenclatures in the area of laboratory test names (LOINC) and therapeutic drugs (Multum's MediSource Drug Lexicon), in order to achieve an integrated whole that solves the problem of a clinical reference terminology. PMID- 10384526 TI - Nursing terminology: a comparison of the ICNP and the nursing intervention lexicon and taxonomy. AB - The purpose of this paper is to provide an overview of nursing terminology work done to date and to compare the labels and subsumed terms of the recent alpha version of the International Classification of Nursing Practice (ICNP) with the verb terms (n = 147) of the interventions in a dataset of interventions (n = 7292) categorized using the Nursing Intervention Lexicon and Taxonomy (NILT). Two estimates were used to evaluate the adequacy of the ICNP terms for representing intervention terminology. Term matches were done using the NILT categories most similar to the ICNP action types. The ICNP action type 'observing' (and its subsumed terms) was best, accounting for 20% of the NILT verbs. The ICNP label 'observing' (and its subsumed terms) ranked first in accounting for 69% of the interventions in the NILT categories (CND & CV). The remaining action types had many fewer matches for the verbs and interventions in the NILT dataset. Thus it is possible to conclude that a relatively small subset of the verb terms, and interventions in a dataset of natural language interventions categorized using NILT have been captured by the INCP alpha version of Axis A. PMID- 10384527 TI - An evaluation of UMLS as a controlled terminology for the Problem List Toolkit. AB - We are developing a set of software components--the Problem List Toolkit (PL-Tk)- to support operations on clinical problem labels. An adaptation of the National Library of Medicine's Unified Medical Language System (UMLS) provides general vocabulary services to domain-specific software components. Our initial investigation centers on the inclusion in UMLS of problem labels used in the Beth Israel Deaconess Medical Center's Online Medical Record (OMR). We also explore the semantic typing of problem labels matched in UMLS. We have operationally defined a clinical problem to derive its semantic type from classes of terms representing findings or processes typically requiring diagnostic evaluation or therapeutic management in clinical practice. Of 1262 unique OMR problem labels, 999 terms (79%) have matches in UMLS. 986 of 999 terms (99%) map to the UMLS concept of the corresponding lexical match. 952 of 999 terms (95%) have semantic types that comply with our operational definition of clinical problems. These 952 terms (75%) constitute Version 1.0 of the problem list vocabulary B196. Matching terms with inappropriate semantic types raise issues regarding requirements for PL-Tk, typing of existing UMLS terms, and the adequacy of our operational definition for clinical problems. UMLS provides a large repertoire of pre coordinated terms that are used as problem labels in a heavily used computer based patient record system. The semantic type hierarchy provides a framework for the consistent use of clinical concepts in problem lists such that clinical problem labels represent "good" clinical problems. PMID- 10384528 TI - Semantic modeling of a traditional classification: results and implications. AB - A primary health care version of the International Statistical Classification of Diseases and Related Health Problems, Tenth Revision (ICD-10), together with a three-dimensional model for classification of diseases according to location, origin, and type has been semantically represented. The resulting computer-based version is made available via the World Wide Web. PMID- 10384529 TI - A concept model for the automatic maintenance of controlled medical vocabularies. AB - A controlled medical vocabulary is a fundamental requirement in a range of medical informatics applications. Large vocabularies development and maintenance is labor intensive and costly. Maintainers of medical vocabularies need appropriate tools to do their work correctly. In this paper, we describe our concept model for a controlled medical vocabulary. We present how this model can check vocabulary consistency. We propose a set of tools in a distributed environment, which permits edition, visualization and maintenance of medical terminologies. PMID- 10384530 TI - Galen-In-Use: using artificial intelligence terminology tools to improve the linguistic coherence of a national coding system for surgical procedures. AB - GALEN has developed a language independent common reference model based on a medically oriented ontology and practical tools and techniques for managing healthcare terminology including natural language processing. GALEN-IN-USE is the current phase which applied the modelling and the tools to the development or the updating of coding systems for surgical procedures in different national coding centers co-operating within the European Federation of Coding Centre (EFCC) to create a language independent knowledge repository for multicultural Europe. We used an integrated set of artificial intelligence terminology tools named CLAssification Manager workbench to process French professional medical language rubrics into intermediate dissections and to the Grail reference ontology model representation. From this language independent concept model representation we generate controlled French natural language. The French national coding centre is then able to retrieve the initial professional rubrics with different categories of concepts, to compare the professional language proposed by expert clinicians to the French generated controlled vocabulary and to finalize the linguistic labels of the coding system in relation with the meanings of the conceptual system structure. PMID- 10384531 TI - Lessons learned from co-operative terminology work in the medical domain. AB - High-quality terminologies are crucial for communication, documentation, and information retrieval. The creation, adoption, and maintenance of such terminologies is a complex task that requires human co-operation. We have developed a terminology server that supports remote, asynchronous co-operation and allows data inconsistencies that can later be resolved through human discussion. We have employed the terminology server in two projects and report on the lessons learned, which have led us to extend our approach. PMID- 10384532 TI - Why discourse structures in medical reports matter for the validity of automatically generated text knowledge bases. AB - The automatic analysis of medical full-texts currently suffers from neglecting text coherence phenomena such as reference relations between discourse units. This has unwarranted effects on the description adequacy of medical knowledge bases automatically generated from texts. The resulting representation bias can be characterized in terms of artificially fragmented, incomplete and invalid knowledge structures. We discuss three types of textual phenomena (pronominal and nominal anaphora, as well as textual ellipsis) and outline basic methodologies how to deal with them. PMID- 10384533 TI - Use of project ontologies and terminology servers to support software engineering. AB - Complex medical software imposes new requirements on the methods and tools used for maintenance. Appropriate maintenance tools can increase software reliability and quality by providing means to trace dependencies among software artifacts for reducing unexpected impacts in software caused by software changes. We have used the GRAIL concept-representation language for medical terminologies to build a project ontology that models relationships among software artifacts. Our approach involves modeling of the terminology used in software projects, which enables us to describe, classify and relate individual software artifacts. A networked repository accessible to the entire software development staff stores the conceptual model, source code and associated documents. We present an architecture for a maintenance tool, and show how developers can use GRAIL to build a project ontology. PMID- 10384534 TI - VM-in-Protege: a study of software reuse. AB - Protege is a system that encompasses a suite of graphical tools and a methodology for applying them to the task of creating and maintaining knowledge-based systems. One of our key goals for Protege is to facilitate reuse on new problems of components of previously developed solutions. We investigated this reusability by applying preexisting library components in the Protege system to a reconstruction of VM, a well-known rule-based system for ventilator management. The formal steps of the Protege methodology-ontology creation, problem-solving method selection, knowledge engineering, and mapping-relation instantiation-were followed, and a working system with much of the reasoning capability of the original VM was created. The work illuminated important lessons regarding aspects of component reusability. PMID- 10384535 TI - Automated acquisition of rules from clinical databases and its evaluation. AB - This paper presents an approach to induction of rules from databases using rough set model. The system was evaluated on three clinical databases, and induced results were compared with other conventional rule induction methods and medical experts' rules. The results show that the introduced results outperforms other methods, but that the description length of induced rules is a little short, compared with that of experts' rules, which suggests that experts' rules are combination of different kinds of reasoning, rather than simple classification. PMID- 10384536 TI - Building cross-thesauri with the support of UMLS. AB - The absence of a robust system of descriptors (cross-thesaurus) hampers the development of combinatorial terminological systems. We developed a tool (I BROWSE) to produce a cross-thesaurus by analyzing terminological corpora. To facilitate the work of experts and to produce re-usable results, our application interacts via the Internet with the UMLS Knowledge Sources Server. We applied our tool on 2999 dissections on surgical procedures produced in the project GALEN-IN USE, as a part of the internal Quality Assurance program. Support from UMLS seems mostly promising about descriptors on , , and . Additional assistance can be given to domain experts on less frequent descriptors on pervasive modifiers. We plan to apply our tool also to production of terminological standards in CEN, as a part of a worldwide process of gradual convergence and transformation of coding systems into second-generation systems and terminological services. PMID- 10384537 TI - The role of compositionality in standardized problem list generation. AB - Compositionality is the ability of a Vocabulary System to record non-atomic strings. In this manuscript we define the types of composition, which can occur. We will then propose methods for both server based and client-based composition. We will differentiate the terms Pre-Coordination, Post-Coordination, and User Directed Coordination. A simple grammar for the recording of terms with concept level identification will be presented, with examples from the Unified Medical Language System's (UMLS) Metathesaurus. We present an implementation of a Window's NT based client application and a remote Internet Based Vocabulary Server, which makes use of this method of compositionality. Finally we will suggest a research agenda which we believe is necessary to move forward toward a more complete understanding of compositionality. This work has the promise of paving the way toward a robust and complete Problem List Entry Tool. PMID- 10384538 TI - Automatic prediction of trauma registry procedure codes from emergency room dictations. AB - Current natural language processing techniques for recognition of concepts in the electronic medical record have been insufficient to allow their broad use for coding information automatically. We have undertaken a preliminary investigation into the use of machine learning methods to recognize procedure codes from emergency room dictations for a trauma registry. Our preliminary results indicate moderate success, and we believe future enhancements with additional learning techniques and selected natural language processing approaches will be fruitful. PMID- 10384539 TI - From French vocabulary to the Unified Medical Language System: a preliminary study. AB - The Unified Medical Language System (UMLS) is an extensive source of biomedical knowledge developed and maintained by the U.S. National Library of Medicine (NLM). The UMLS began to include biomedical terms in other languages a few years ago. However, providing foreign terms for existing concepts is only the first step for the UMLS to become international. The current limits of the use of the UMLS in French are analyzed (partial translation, unique source of the translated concepts, improper character set, and absence of lexical resources for lexical matching tools). Some suggestions are given for French to be better integrated into the UMLS, especially for adapting the lexical resources to French. Once completed, our present work is expected to give the UMLS the capability to be effectively queried in French. PMID- 10384540 TI - Medical diagnostic system using Fuzzy Coloured Petri Nets under uncertainty. AB - We propose a medical diagnostic system using Fuzzy Coloured Petri Nets (FCPN) in this paper. For complex real-world knowledge Fuzzy Petri Net (FPN) models have been proposed to perform fuzzy reasoning automatically. However, in the Petri Net we have to represent all kinds of processes by separate subnets even though the process has the same behavior of other one. Real-world knowledge often contains many parts which are similar, but not identical. This means that the total PTN becomes very large. The kind of problems may be annoying for a small system, and it may be catastrophic for the description of large-scale system. To avoid this kind of problems we propose a learning and reasoning method using FCPNs under uncertainty. On the other hand to correct the rules of knowledge-based system hand-built classifier and empirical learning method both based on domain theory have been proposed as machine learning methods, where there is a significant gap between the knowledge-intensive approach in the former and the virtually knowledge-free approach in the later. To resolve such problems simultaneously we propose a hybrid learning method which is built on the top of knowledge-based FCPN and Genetic Algorithms (GA). To verify the validity and the effectiveness of the proposed system, we have successfully applied it to the diagnosis of intervertebral diseases. PMID- 10384541 TI - Barriers to evaluation of clinical vocabularies. AB - BACKGROUND: The importance of standard vocabularies for representing clinical data is widely accepted. The selection of suitable vocabularies for a given task relies upon criteria for systematically differentiating vocabularies. However, methods and measures for comparing vocabularies are only now emerging. Substantial barriers inhibit progress in the development of these measures. METHOD: As part of a larger project, the authors are developing quantitative measures for characteristics of controlled vocabulary as identified in the literature. RESULTS: Seven barriers have been identified that inhibit the development of quantitative measures of the characteristics of controlled vocabularies: 1) application dependence; 2) empirical v. independent assessment; 3) dichotomous v. continuous measures of characteristics; 4) poor definition of characteristics; 5) number of characteristics; 6) multiple levels of significance of characteristics; and 7) interdependence of characteristics. CONCLUSION: Progress toward quantitative assessment of clinical vocabularies is dependent upon overcoming barriers to the development of appropriate measures. Such progress requires innovative solutions and collaboration among investigators. PMID- 10384542 TI - Medical language processing applied to extract clinical information from Dutch medical documents. AB - In this paper, we want to show how an existing morpho-syntactic analyser for Dutch (Dutch Medical Language Processor--DMLP) has been extended in order to produce output that is compatible with the language independent modules of the LSP-MLP system (Linguistic String Project--Medical Language Processor) of the New York University. The former can focus on idiosyncrasies for Dutch and take advantage of the language independent developments of the latter. This general strategy will be illustrated by a practical application, namely the extraction of clinical information from Dutch patient discharge summaries. Such an application can be of use for education, research and quality control purposes in a hospital environment. PMID- 10384543 TI - Land of morning calm. Past and future. PMID- 10384544 TI - Administrators, doctors or patients. Who are the users of information system. PMID- 10384546 TI - Health informatics in the Western Pacific Region of WHO--current issues and concerns. PMID- 10384545 TI - The evolution of health-care records in the era of the Internet. AB - Health-care records are evolving as the rationale for their automation becomes ever more persuasive. Promoting that evolution are local and wide-area networking as they demonstrate that computer-based medical and health records will be more than mere electronic versions of the traditional paper record. System integration has emerged as a key element in the reinvention of environments for patient data management and health promotion. The ability to achieve the future vision of integrated health records depends in part on current research initiatives related to the role of the global information infrastructure in supporting health and health care. PMID- 10384547 TI - From electronic medical record to personal health records: present situation and trends in European Union in the area of electronic healthcare records. AB - In this article we define the electronic healthcare record (EHR) and present its purpose as a tool for continuity of care. We consider the EHR system as a necessary tool for collaborative work of healthcare professionals linking the traditional stand alone physician's systems or departmental systems, which we refer to here as electronic medical record systems. We briefly describe the current usage of electronic medical records in EU and focus on the major challenges to wide implementation of electronic healthcare record systems. Finally, we point out trends that show stronger involvement of patients (citizens) in the health care process and discuss the impact on future EHR systems. We call the next generation of EHR that takes into account the new role of citizens Personal Health Records. PMID- 10384548 TI - Issues facing system vendors as we approach the 21st century and expected roles- implementing a hospital information system providing greater patient satisfaction. PMID- 10384549 TI - Building patient workflow management systems by integrating medical and organizational knowledge. AB - Patient management is a distributed activity involving general practitioners, clinicians, analysts, nurses, etc. Thus an integrated Patient Workflow Management System (WfMS), based on a detailed model of both the organizational and medical knowledge, could heavily improve Health Care System's performance in terms of collaborative work and resource utilization. A set of tools was developed to improve 1) acquisition of medical knowledge represented through clinical practice Guideline, and 2) acquisition of organizational knowledge describing the work process. PMID- 10384550 TI - Using informatics principles and tools to harness research evidence for patient care: evidence-based informatics. AB - With the huge worldwide investment in biomedical research during the past 50 years, there are many important advances in health care knowledge each year. Unfortunately, it commonly takes over 20 years for even the most important of these advances to be widely integrated into clinical practice. Many potentially remediable factors are responsible for this dilemma in research transfer, including defective continuing education for health professionals and patients; increasingly complex medical regimens; diminishing resources for health care; and inadequate evidence management. The principles and procedures of health informatics can help overcome some of these barriers to research transfer, particularly such evidence management tasks as retrieving, processing, summarizing, disseminating and applying evidence for clinical care. Evidence retrieval has been improved by better indexing and electronic search engines, by improved access from clinical and other settings, and by integration of evidence into clinical decision support systems. Evidence processing has been greatly accelerated by streamlined methods of critical appraisal of research and by centralization of these procedures for the development of current awareness publications and cumulative "best evidence" databases. The Cochrane Collaboration has revolutionized the summarization (systematic review) of evidence. The internet has provided access to patients, practitioners, and policy makers, alike. Direct-from-patient automated data collection promises to move the connection between evidence and practice to a higher level. In all of these innovations, health care practice is most likely to be enhanced by intertwining best evidence with best informatics techniques. PMID- 10384551 TI - Virtual electronic patient records for shared care. AB - Systems that primarily serve health-care organizations are changing into systems that support patient care. The core of this change is shaped by systems for computer-based patient records (CPRs), which are part of local or regional networks, giving access to data in different information systems. In principle, it should not matter where the patient data are located as long as data can be transferred to the physical location where patients and clinicians meet. Networking and electronic communication enable to realize an environment that makes all systems where patient data reside, acting as one integrated, virtual CPR-system from the user's perspective. The patient record itself needs not to be physically located at one place, but may be virtual. A development in this direction is the European 14C project, which aims at integrating patient record data, images, and biosignals from whatever system they are stored and on whatever computer they are processed in the network. PMID- 10384552 TI - The economic implications of users willingness to increase knowledge capital in health informatics. AB - OBJECTIVE: To develop an economic model of health care professionals demand for knowledge capital in health informatics. DESIGN: Case study with application of the Contingent Valuation Method to develop a small-scale model. SETTING: Specialized clinic at a university Hospital in Sweden. RESULTS: The model displays the economic rationale behind an individual's choice to spend leisure time for obtaining knowledge in health informatics. This decision reduces the total leisure time, but does not increase salary. Instead, it may increase the personal well being by higher satisfaction gained from using information systems and by being recognized as a computer expert. CONCLUSIONS: Individuals have preferences over all uses of time and for activities they can choose to engage in Support of health care staff's investment in health informatics knowledge capital may benefit both the individuals and indirectly the health care organization. PMID- 10384553 TI - Educational benefits and cost savings from an interactive multimedia "best practice". AB - BACKGROUND: Developing and deploying "clinical practice guidelines" and "best practices" is a major strategic goal of our organization. We developed and evaluated a novel way of educating our colleagues on the office management of shoulder pain, using an interactive multimedia computer program to allow rapid access to user-selected audiovisual educational materials through a simple point and click interface. METHODS: Pre- and post-testing of 30 practicing primary care providers was conducted to assess ability to evaluate and manage patients who present with shoulder pain, and a post-use survey was administered to assess clinician acceptance and confidence, ability to use as a just-in-time tool, and changes in utilization of medical resources. RESULTS: Pre- and post-testing demonstrated that clinicians used this tool to improve their scores in a wide variety of clinical areas including history, physical examination, diagnosis, and management of conditions causing painful shoulder above their scores using textbooks and other traditional educational materials alone. The post-use survey showed outstanding acceptance by clinicians and their reported increased confidence as well as the potential for their increased abilities to translate into significant cost savings through more focused use of radiographic studies, orthopedic consultations, and physical therapy treatments. CONCLUSIONS: It was concluded that "The Multimedia Reference for Office Management of Shoulder Pain" is an effective and valuable learning resource for practicing clinicians, and demonstrates the potential to use an interactive multimedia tool to augment clinical skills and promote cost effective management of patients with this common clinical problem. PMID- 10384554 TI - Methodology for developing educational hypermedia systems. AB - Hypermedia has the potential to greatly enhance teaching. It can, however, be difficult to develop hyperdocuments which provide medical students with the full benefit of the technology. We wish to resolve this problem by providing a methodology for creating educational hypermedia systems with specific emphasis on the medical domain, but applicable across disciplines. This paper examines some of the specific issues involved in educational hypermedia, and outlines a series of practical guidelines, drawing together the disparate disciplines necessary to the development process. We illustrate the methodology with our own experience in creating and updating the HIV Hypermedia Medical Education Software over a period of two years. PMID- 10384555 TI - Lifestyle evaluation system to support health education. AB - A computer-based system for evaluation of one's lifestyle was designed so that public health nurses could use it for health education to improve a patient's lifestyle and to prevent lifestyle-related diseases. The Lifestyle Evaluation System (LES) is a computer program that works on a personal computer. The LES is consist of four parts; inputting personal data, answering the questionnaire, showing the result and getting health check up data. The questionnaire includes 40 questions regarding diet, smoking, drinking, physical activity, rest, social activity and health care activities, based on Breslow's seven lifestyles. The result offers participants' lifestyle evaluation in forms of values, graphs, tables and messages. Evaluation values are deviation value, rank, BMI and standard weight. The LES also offers participant's periodic health check data, average data derived from all residents' data, and past results of LES so that public health nurses are able to use these for health education. The health check data is obtained from another database stored on floppy disks. The participants input data by themselves and get result immediately. The interactive style is effective in raising interest in health education. PMID- 10384556 TI - Using multimedia virtual patients to enhance the clinical curriculum for medical students. AB - Changes in the environment in which clinical medical education takes place in the United States has profoundly affected the quality of the learning experience. A shift to out-patient based care, minimization of hospitalization time, and shrinking clinical revenues has changed the teaching hospital or "classroom" to a degree that we must develop innovative approaches to medical education. One solution is the Virtual Patient Project. Utilizing state-of-the-art computer based multimedia technology, we are building a library of simulated patient encounters that will serve to fill some of the educational gaps that the current health care system has created. This project is part of a newly formed and unique organization, the Harvard Medical School-Beth Israel Deaconess Mount Auburn Institute for Education and Research (the Institute), which supports in-house educational design, production, and faculty time to create Virtual Patients. These problem-based clinical cases allow the medical student to evaluate a patient at initial presentation, order diagnostic tests, observe the outcome and obtain context-sensitive feedback through a computer program designed at the Institute. Multimedia technology and authoring programs have reached a level of sophistication to allow content experts (the teaching faculty) to design and create the majority of the program themselves and to allow students to adapt the program to their individual learning needs. PMID- 10384557 TI - Development of a Web-based CAI program for maternity nursing practice. AB - This quasi-experimental study was conducted to explore what kind of impact a Web based educational program can have on nursing students and how nursing students react to this educational program. A Web-based computer assisted instruction for maternity nursing practice was developed by researchers using Instruction System Design model and serviced for nursing students on the WWW. To study educational effect of this program on nursing students 30 senior students at a school of nursing were selected. They were assigned into experimental and control groups based on their maternity nursing practice schedule. The effect of the Web-based CAI was measured using Achievement score, attitude toward learning experience and attitude toward computer and Internet, and compared between two groups. There were significant differences between two groups in achievement and attitude score. And students in the experimental group showed positive response to the Web based CAI program. These results suggest that Web-based CAI is useful as a new teaching tool for maternity nursing practice as well as other nursing courses. Comments on program improvement and operational issues were collected from students. These will be used for program improvement in future. PMID- 10384558 TI - Problem oriented project work in a distance education program in health informatics. AB - At Aalborg University, an important part of the distance education program within Health Informatics is problem oriented project work. Traditionally, distance education has been characterized by one-way communication and self study whereas the problem oriented project study form is based on cooperation and dialogue. In this paper, we describe the way in which we have implemented the problem oriented study form in a program within Health Informatics which is based on distance learning. First, we describe the program with regard to student, structure, aim, and activities. Second, we introduce the problem oriented project study form and present the basic principles behind this approach. Third, we explain important concepts and distinctions within the area of distance education. Finally, we describe the way in which we try to put the ideals of the problem oriented project work into practice. The use of a computer conferencing system is essential but in our experience, it is not in itself sufficient to provide the necessary support for the student project work. PMID- 10384559 TI - WEB-MO--a computer aided learning on WWW. AB - The paper presents an electronic textbook on World Wide Web (WWW), which present theoretical notions about the eyes' movements. It also contains clinical cases as example for the described notions. The application is being tested at the County Hospital, Department of Neurosurgery, Timisoara, Romania. The reaction of the educators and learners is quite positive. We think there will be a great learning tool for the future. PMID- 10384560 TI - Setting a national informatics agenda for nursing education and practice to prepare nurses to develop and use information technology. AB - Efforts to build a telecommunications infrastructure that will link people nationally and internationally includes a focus on health care. A telecommunications infrastructure will add value to providers and consumers of health care only if they are able to use it to access and share information. Nurses often have information technology available to them, but are inadequately prepared to use it. Under the direction of the National Advisory Council for Nursing Education and Practice (NACNEP), the Division of Nursing convened a panel of experts in nursing informatics to recommend an informatics agenda. Using a nominal group technique the informatics experts identified informatics needs and recommended initiatives that would better prepare nurses to use and develop information technology. Their recommendations were reviewed by NACNEP resulting in a national informatics agenda for nursing education and practice. PMID- 10384561 TI - Innovative informatics education: aligning theory and practice through strategic partnership. AB - The University of Victoria School of Health Information Science (HINF) and The Toronto Hospital (TTH) have formed a strategic partnership to align current information management theory with practical experience. The fourth year undergraduate course in Information Management and Technology was redesigned to include a joint research project with TTH. Staff from TTH were paired with groups of 2-3 students to explore current information management issues. Technology such as electronic mail and conference calls enabled timely communication. Final results were presented via video conference at the end of the term and jointly graded by the professor and hospital staff. Over the three years, a variety of projects have been undertaken and the approach has been refined. Each year offered new insight to both students and staff. Findings include several key success factors: senior level support, early planning, timely communication, access to technology, and opportunity for staff and student feedback. This partnership offers strategic advantages to both organizations in innovation, mutual learning, recruiting, and partnership. Next steps include expansion of the project to a regional model and seminars on current information management topics for TTH staff. PMID- 10384562 TI - Twenty care components: an educational strategy to teach nursing science. AB - The HHCC System's twenty care components and nomenclature provide a framework for the development of teaching strategies to prepare nurses for practice in the 21st century. The use of an integrated system of instruments accessible to students on line allows for a smooth transition from course to course and later to clinical nursing practice. Faculty and/or instructors at Georgetown University School of Nursing use this innovative educational strategy and conceptual approach to facilitate collaboration with other healthcare providers and to model professional behavior for students. PMID- 10384563 TI - Assessing the use of low cost PC-based ISDN videoconferencing in hospital training. AB - A course on Italian safety rules and regulations has been delivered to the personnel of a University clinic using videoconferencing, based on an ISDN-2 connection through a multimedia PC. The goal was to evaluate the efficacy of this technology for the training of hospital personnel. To assess it, the trainees were divided into two groups, one at the trainer's site and the other in the University. Comparing the evaluating questionnaires that both groups were asked to complete at the end of each session (based on the comprehension of the subject taught), significant differences were found only when the quality of images was very poor (for technical reasons) or the level of interaction was low. The price/performance ratio was considered good, but there are some technical and psychological problems that must be taken in account before using videoconferencing for training. PMID- 10384564 TI - Collaboration over the Internet--the Melbourne-Singapore way. AB - The University of Melbourne and The National University of Singapore have established a joint WWW page under the auspices of the Cyberspace Hospital to facilitate inter-regional collaboration and discussion about best practice in research, teaching and clinical activities in general and family practice. Information about each of the two departments are available as well as an electronic forum over the WWW (URL is http:/(/)ch.nus.sg/fammed). Seminal papers on research, teaching and clinical practice are discussed and summarised after critique from GPs from both regions. A paper is discussed every 8 weeks: 1 week to post the paper plus a critique, 6 weeks for discussion, and 1 week to sum up and post the summary. It is planned to progress to audio- and video-conferencing and to widen the participant groups. The process of and difficulties with establishing, maintaining and running the electronic forum is described. Results of the discussions, comments from participants and strategies for the future will be presented as well as a demonstration of the electronic forum. PMID- 10384565 TI - Model of computerized academic medical clinic. AB - In the traditional medical graduation course, the student receives a great amount of information while training at the Outpatient Care; the student assumes the physician's role, collecting all the information regarding the patient's clinical history and learns to get along with patients as well. During the attendance process, several factors interfere in the academic teaching, such as limitations of room numbers, amount of patients, difficulties in obtaining medical records, paper illegibility, among other problems. Due to those difficulties, the Model of Computerized Academic Health Clinic, implies in a new learning paradigm in the medical practice, rethinking the traditional process of learning-attendance, where the old model, in which attendance is restricted to a place, is extended in an open atmosphere of shared knowledge, rich of computer resources. The pilot project was implanted in the Pediatrics General Health Clinic of UNIFESP/EPM. It allows that the fifth-year medical students, residents, trainees and tutors use computerized clinics, connected with the academic net of UNIFESP and to the Internet. All the computing and information resources settled at the Outpatient Care improved the organization of its services, increased the medical students' curiosity, improved their participation in learning through interactive programs and clinical attendance. PMID- 10384566 TI - A virtual university Web system for a medical school. AB - This paper describes a Virtual Medical University Web Server. This project started in 1994 by the development of the French Radiology Server. The main objective of our Medical Virtual University is to offer not only an initial training (for students) but also the Continuing Professional Education (for practitioners). Our system is based on electronic textbooks, clinical cases (around 4000) and a medical knowledge base called A.D.M. ("Aide au Diagnostic Medical"). We have indexed all electronic textbooks and clinical cases according to the ADM base in order to facilitate the navigation on the system. This system base is supported by a relational database management system. The Virtual Medical University, available on the Web Internet, is presently in the process of external evaluations. PMID- 10384567 TI - Curriculum development and courseware implementation using World Wide Web technology. AB - The curriculum development of medical informatics related courses and their adoption into the faculty curriculum is one of the main tasks of the present day educators. To this end the Internet technology has supported this task. The Internet was born in December of 1969 and has grown phenomenally since. Its graphically interactive, user-friendly modality, the World Wide Web (WWW), is younger and growing even more explosively. By its nature, the WWW is a tool ideally and uniquely suited for the advancement education. This paper describes the design, development and the implementation of a Web Site for supporting the education of the students in the Faculty of Nursing at the University of Athens. The application will be used also in the European project Nightingale. PMID- 10384568 TI - Investigating a concept-based tool to enhance the process of curriculum development: an example for healthcare education. AB - This paper reports on a first exploration carried out in the CoMET (COncept Modelling Environment for Teachers) project to investigate the educational potential of a concept-based toolkit developed in the GALEN project. In particular, on whether such tools can offer a basis to support and enhance healthcare curriculum development. We first adapted part of the toolkit by developing a front-end user friendly interface to access the conceptual knowledge. We then collected data from an empirical study. Preliminary results suggest that the concept-based approach of the CoMET tools is viable and has several potential benefits for educational activities. Directions for further work in the use of the concept-based tool for teaching as well as learning are also discussed. PMID- 10384569 TI - The development of digital library system for drug research information. AB - The sophistication of computer technology and information transmission on internet has made various cyber information repository available to information consumers. In the era of information super-highway, the digital library which can be accessed from remote sites at any time is considered the prototype of information repository. Using object-oriented DBMS, the very first model of digital library for pharmaceutical researchers and related professionals in Korea has been developed. The published research papers and researchers' personal information was included in the database. For database with research papers, 13 domestic journals were abstracted and scanned for full-text image files which can be viewed by Internet web browsers. The database with researchers' personal information was also developed and interlinked to the database with research papers. These database will be continuously updated and will be combined with world-wide information as the unique digital library in the field of pharmacy. PMID- 10384570 TI - An international dermatological image atlas on the WWW: practical use for undergraduate and continuing medical education, patient education and epidemiological research. AB - We describe the development of an image database DOIA (Dermatological OnlIne Atlas) and present several spin-off projects using images of the atlas, e.g. student education using the atlas including results of an questionnaire evaluating computer-literacy, prerequisites and interests of students for using computers and the World-Wide-Web (WWW), a patient information system and an experiment to collect epidemiological data from patients with dermatological diseases via WWW. The database, available on the WWW at http:@www.derma.med.uni erlangen.de, contains about 3,000 clinical images covering more than 540 dermatological diagnoses. It is designed for worldwide use; international submissions are encouraged. One aim of the project is to compile an international reference for dermatological images, containing images of high educational quality and also covering conditions on different skin types and rare diagnoses which are not commonly illustrated in ordinary textbooks. All images were originally mapped to the Erlanger Diagnosis Code, which is a proprietary modified ICD-9 key, later also to the UMLS (Unified Medical Language System). In addition, images are described with keys for the location, physical attributes of the location and clinical and histopathological features of the lesion. In order to facilitate the integration of the atlas into other web-based medical resources and to allow easy access to additional information, the Erlanger Diagnosis Code was mapped to the CUIs (unique concept identifiers) of the UMLS Metathesaurus. One purpose of the UMLS is to allow conversion of terms from one controlled medical vocabulary to another, thus, mapping of our diagnosis code to the UMLS CUIs allows simultaneous search for a given diagnosis in a number of other databases and also access to our image database from other databases. Mapping was successful for 619 out of 1383 dermatological diagnosis terms. For images with these diagnoses we are able to provide a hyperlink to other databases available on the Internet, such as MEDLINE, PDQ and OMIM, with automatic retrieval using the preferred vocabulary of the respective database. By grouping all diagnoses into sets with similar morphologies we further integrated a differential diagnosis mode. In order to educate patients via the Internet, a separate patient information system has been developed, using images of the electronic atlas. As an experiment to explore the feasibility of the Internet to gather epidemiological data from patients, users are asked to complete an electronic questionnaire covering signs for atopy. We conclude that an online image atlas has multiple educational, clinical and research applications. PMID- 10384571 TI - Concepts of a Web-based open distributed textbook for the multimodal diagnostics of gastrointestinal tumours with MRI, CT and video-endoscopy addressing students of medicine and students of medical informatics as two different target groups. AB - Multimodal diagnostics of gastrointestinal tumours with MRI, CT and video endoscopy is a rapidly changing domain. The education at our universities should overcome the obstacles of traditional learning based on paper media and oral lectures with retention rates of 10-30% only. The paper presents the objectives and the results of the design phase of the project ODITEB1-Open Distributed TExt Book, for Computer-Assisted Instruction in the domain mentioned above. The main objective is to produce an electronic interactive textbook in order to shift education to more efficient learning settings with higher retention rates. The main concepts are 1) three-layer architecture (dynamic case layer, intermediate query layer, static instruction layer) 2) case pool distribution 3) active learners experience (interactive exploration of original image data). PMID- 10384572 TI - Impact of the TOP-FORUM hypermedia system in a pediatric oncology care unit. AB - This paper describes our approach in analyzing the impact of the TOP-FORUM hypermedia in a pediatric oncology care unit. The impact of this technology is realized through the study of accommodation and assimilation adoption. Accommodation refers to the technological adoption and Assimilation refers to the professional adoption. Results show that accommodation depends on information and formation of the users. Assimilation is more difficult to evaluate because it depends on human, social and organizational problems. PMID- 10384573 TI - Measuring and improving quality using information systems. AB - Information systems (IS) are increasingly important for measuring and improving quality. In this paper, we describe our integrated delivery system's plan for and experiences with measuring and improving quality using IS. Our approach is that for quality measurement to be practical, it must be integrated with the routine provision of care, and whenever possible should be done using IS. Thus, at one hospital, we now perform almost all quality measurement using IS. However, IS are not only useful for measuring care, but represent powerful tools for improving care using decision support. Specific areas in which IS has already been particularly effective include reducing the unnecessary use of laboratory testing, reporting important abnormalities to key providers rapidly, adverse drug event detection and prevention, initiatives to reduce the costs of drugs, and making critical pathways available to providers. The next wave of effort will be to promote widespread use of computerized guidelines, which is likely to prove more challenging. However, the advent of managed care in the U.S. has produced strong incentives to provide high quality care at low cost, and our perspective is that only with better IS than exist today will this be possible on a widespread basis. Such systems make feasible implementation of care improvement and cost reduction initiatives on a scale which could not previously be considered. PMID- 10384574 TI - Computer-adjusted dosage of anticoagulant therapy improves the quality of anticoagulation. AB - OBJECTIVE: Risks and benefits of anticoagulant therapy depend directly of the quality of anticoagulation. We performed a meta-analysis of published randomized trials to assess the overall effectiveness of computer-based prescription systems on the quality of anticoagulation. DESIGN: Randomized controlled trials were identified through electronic searches of the Medline database (1966-1997) and systematic analyses of the references of articles. Two investigators selected relevant papers and summarized data from the studies. METHODS: The outcome variable was the proportion of days within the target range of anticoagulation. A pooled estimate of the common odds ratio of being in the target range and its confidence interval was obtained by the Mantel-Haenszel method. RESULTS: Seven trials having included 1217 patients were identified. Computer systems were based on a pharmacokinetic-pharmacodynamic model and a bayesian prediction method. Most of them concerned the oral anticoagulant warfarin. The global odds ratio of being in the target range was 1.58 [95% CI: 1.34-1.86], thus meaning that the use of a computer for anticoagulation optimization increased by 58% the proportion of visits where patients were appropriately treated. The proportion of clinical events was too low for allowing a summary analysis. CONCLUSION: Evidence from randomized controlled trials supports the effectiveness of computer-aided anticoagulant prescription. Diffusion of these systems in ambulatory care could increase the benefit/risk ratio of anticoagulant treatment at a low cost. PMID- 10384576 TI - Siegfried: System for Interactive Electronic Guidelines with Feedback and Resources for Instructional and Educational Development. AB - The proliferation of clinical practice guidelines (CPGs) has necessitated computerized solutions for guideline distribution and implementation. In this paper we describe a Web-based system that interactively presents CPGs at the point of care. Our system, known as Siegfried, provides a generalized solution for implementing CPGs by maintaining the guideline knowledge base separate from the application that presents the guidelines. As a result of this design, new CPGs can be easily added and existing CPGs can be expeditiously modified without additional programming. This system also solicits feedback from users regarding guideline recommendations and provides hypertext links to relevant Web-based instructional and educational resources. PMID- 10384575 TI - Continuous quality improvement through use of perinatal clinical record and AGUSTINA database. AB - This paper reviews the development of the field of medical informatics in Latin America. It also describes the results of a computer-based data management system, named AGUSTINA, which is comprised of maternal and infant data on 7570 deliveries that occurred between June, 1990 and December, 1996 in a hospital in the surroundings of Buenos Aires, Argentina. These data were fundamental for the instrumentation of preventive community-oriented activities in the area. Finally, this paper describes recommendations for future actions in the area of medical informatics in Latin America. ABBREVIATIONS: PCR: perinatal clinical record; CLAP, Centro Latino-Americano de Perinato-logia; WHO, World Health Organization; PAHO, Pan-American Health Organization; E-Mail, electronic mail; WWW, world wide web; FTP, file transfer protocol. PMID- 10384577 TI - Activity modelling for assessing the usability of telematics applications in healthcare. AB - With the development of Telematics Applications in Healthcare, it becomes obvious that the success of these services is tightly linked with their usability and their acceptance by users. That is why Users Requirements Analysis appears as an essential step before implementation of new computer services. In many cases, particularly when doctors are directly involved in the management of information, there is a strong interaction between their usual activity and the new tasks implied by the computer. In those cases, Activity Modelling can be useful to describe the way people are working, the motivations that underlie this activity, the place of information management in this activity. Moreover, these models can help to forecast possible difficulties or reasons for failure of the data management when computerised. Obviously, these models are highly depending on the healthcare organisation but some basic lines can be transferable. In this paper we illustrate the Activity Modelling Methodology by examples from Telematics exchanges between GPs and Hospitals. PMID- 10384578 TI - Review of the new methodology of analyzing receiver operating characteristic curves. AB - The Dorfman and Alf algorithm based on the classic model of signal detection theory is most frequently used for the receiver operating characteristic (ROC) analysis. However, several shortcomings of this method were pointed out. On these problems, in recent years, several authors have proposed the new analysis methods of the ROC curve. This paper reviews these new analysis methods, and discusses them. First, we review the general regression methodology for the ROC analysis, proposed by Tosteson and Begg. Secondly, we review the analysis of variance (ANOVA) method, proposed by Dorfman et al; in this method, the jackknife method was applied to ANOVA for analyzing ROC data. Further, we give the illustrative experimental design, and discuss the application of these methods to our illustrative experiment. We indicate that we can make use of these two methods for the evaluation of diagnostic performance in our illustrative experiment. PMID- 10384579 TI - Present situation and characteristics of Traditional Chinese Medical Informatics (TCMI). AB - This paper provides an introduction to the present basis for the Traditional Chinese Medical Informatics (TCMI). It also analyzes the contents, the characteristics of the information classification of Traditional Chinese Medicine (TCM) and prospects for TCMI. PMID- 10384580 TI - Upgrading clinical decision support with published evidence: what can make the biggest difference? AB - BACKGROUND: To enhance clinical decision support, presented messages are increasingly supplemented with information from the medical literature. The goal of this study was to identify types of evidence that can lead to the biggest difference. METHODS: Seven versions of a questionnaire were mailed to randomly selected active family practice physicians and internists across the United States. They were asked about the perceived values of evidence from randomized controlled trials, locally developed recommendations, no evidence, cost effectiveness studies, expert opinion, epidemiologic studies, and clinical studies. Analysis of variance and pairwise comparisons were used for statistical testing. RESULTS: Seventy-six (52%) physicians responded. On a Likert scale from one to six, randomized controlled clinical trial was the highest rated evidence (mean 5.07, SD +/- 1.14). Such evidence was significantly superior to locally developed recommendations and no evidence at all (P < .05). The interaction was also strong between the types of evidence and clinical areas (P = .0001). CONCLUSION: While most health care organizations present data without interpretation or simply try to enforce locally developed recommendations, such approaches appear to be inferior to techniques of reporting data with pertinent controlled evidence from the literature. Investigating physicians' perceptions is likely to benefit the design of computer generated messages. PMID- 10384581 TI - Professional and organizational impact of using patient care information systems. AB - Evaluation of information systems has often been limited to success factors in relation to system implementation (on time and on budget), training and use of the system. Little attention has been paid to the longer-term effect of using these systems and the resulting issues for health care professionals and organizations. This paper reports on a multiple case study of community hospitals, which examined the impact of using Patient Care Information Systems. An analytic framework incorporated Donabedian's 3 aspects of quality care: structure, process and outcome. These were examined at three levels of impact; direct substitution, proceduralization and new capabilities. Many of the anticipated benefits in the study did not occur because changes occurring in structure (generally taks done by pharmacists and laboratory technologists) did not automatically influence process changes (such as decision making of physicians and nurses) or patient outcomes. Four themes illustrate important professional and organizational issues with implications for worklife of professionals, management and career training. They include the effects of increased efficiency and productivity; "visible" accountability; changing roles and responsibilities; and learning to use new technology versus using new information. PMID- 10384582 TI - The need for a thoughtful deployment strategy: evaluating clinicians' perceptions of critical deployment issues. AB - This paper presents data collected from 899 clinicians across three Department of Veterans Affairs (VA) medical centers where existing terminal-based architecture was being replaced with client-server architecture. Surveys were conducted with physicians (n = 184), nurses (n = 355) and other clinicians (n = 360) gathering user characteristics and their perceptions of five deployment issues (e.g. adequacy of technical and institutional support and perceptions of the soon-to-be implemented clinical workstation). Mean scores for the five deployment issues for all clinicians indicates perceptions are somewhat neutral. However, when data is analyzed according to job classification, significant (p = 0.05) differences in perceptions were noted among groups of clinicians (e.g., physicians and registered nurses). Results of analyzing data grouped by VA site (n = 3) indicates significant (p = 0.05) differences exist among sites in clinicians' perceptions of the deployment issues. A thoughtful deployment strategy including an in-depth assessment of clinician users by job classification and by location may produce important information, critical to the successful deployment of new technologies, in very large health management institutions. PMID- 10384583 TI - The multi-item univariate delta check method: a new approach. AB - The delta check methods are methods for detection of random errors in clinical laboratory tests including specimen abnormalities, specimen mix-up, problems in analysis processes, and clerical errors. Methodologically, it is known that the multivariate delta check methods are more superior to the univariate delta check methods. However, due to some problems in reality including technical difficulties, it is hard to put the multivariate delta check methods into practice. Since the univariate delta check methods are methods at hand, there has been a need for an efficient and effective univariate delta check method. In order to meet such a need, we propose "the multi-item univariate delta check (MIUDC) method". By the multi-item univariate delta check (MIUDC) method, we mean a method in which univariate delta checks are performed on multiple items and specimens with the positive univariate delta check in at least k items are put under a detailed investigation. Our research objectives are the determination of an appropriate value of such k and identification of test items deserving of more interest. Through real data and simulation studies, we concluded that an appropriate value of k is 4 because, with k = 4, we can have light checking-out volumes and high efficiency. Also, we identified total cholesterol, albumin, and total protein as items deserving of more interest because the false positive rate associated with them in the MIUDC was zero in a simulation study. We present the MIUDC method as a quality control method that is easy-to-implement and efficient. PMID- 10384584 TI - Measuring the quality of diagnostic hypothesis sets for studies of decision support. AB - Within medical informatics there is widespread interest in computer-based decision support and the evaluation of its impact. It is widely recognized that the measurement of dependent variables, or outcomes, represents the most challenging aspect of this work. This paper describes and reports the reliability and validity of an outcome metric for studies of diagnostic decision support. The results of this study will guide the analytic methods used in our ongoing multi site study of the effects of decision support on diagnostic reasoning. Our measurement approach conceptualizes the quality of a diagnostic hypothesis set as having two components summed to generate a composite index: a Plausibility Component derived from ratings of each hypothesis in the set, whether correct or incorrect; and a Location Component derived from the location of the correct diagnosis if it appears in the set. The reliability of this metric is determined by the extent of interrater agreement on the plausibility of diagnostic hypotheses. Validity is determined by the extent to which the index generates scores that make sense on inspection (face validity), as well as the extent to which the component scores are non-redundant and discriminate the performance of novices and experts (construct validity). Using data from the pilot and main phases of our ongoing study (n = 124 subjects working 1116 cases), the reliability of our diagnostic quality metric was found to be 0.85-0.88. The metric was found to generate, on inspection, no clearly counterintuitive scores. Using data from the pilot phase of our study (n = 12 subjects working 108 cases), the component scores were moderately correlated (r = 0.68). The composite index, computed by equally weighting both components, was found to discriminate the hypotheses of medical students and attending physicians by 0.97 standard deviation units. Based on these findings, we have adopted this metric for use in our further research exploring the impact of decision support systems on diagnostic reasoning and will make it available to the informatics research community. PMID- 10384585 TI - Heuristic walkthrough evaluation of a prototype computer network service for occupational therapists. AB - AIM: To identify utility and design criteria for a computer network service for occupational therapists. DESIGN: Heuristic walkthrough evaluation. The evaluators explored a prototype and then commented upon the design in plenary sessions. SUBJECTS: One group of information system design experts and one of occupational therapists. RESULTS: The central utility criteria were the possibility to organize dynamic work groups for development of the occupational therapy profession and access to databases on assistive technologies. The main system design criteria identified were navigation, structure, tools, and content. CONCLUSION: A computer network service can support the development of the therapy professions. Generic issues exist which need to be considered in the detailed design. The heuristic walkthrough method is useful for identifying these. PMID- 10384586 TI - Differences of case-mix according to the type of hospital: methodological aspects and results. AB - This study has brought to the fore variations of case-mix according to the type of French hospitals taken into consideration. The GHM line-up in the French classification of the hospital stays (French DRG) have also been studied and variations linked to the type of hospitals have been noticed too. This survey has been carried out thanks to the anonymous discharge summary issued by the national and the regional databases. PMID- 10384587 TI - Evaluation as a tool to increase knowledge in healthcare informatics. AB - The evaluation of information systems is an important topic in Clinical Informatics. It is argued that past evaluations have not been particularly informative in progressing the effective use of IT in healthcare due to their narrow focus. The different roles of evaluation in Clinical Informatics are examined, and the breadth and diversity of the available methodological tool kit highlighted. The aim is to stimulate a greater awareness of the roles and methods of evaluation. Challenges in evaluation which face the Clinical Informatics community are discussed and finally some comments made concerning the way in which evaluation might be made more effective in order to improve our knowledge of how to deliver useful systems into healthcare. PMID- 10384588 TI - Computer experience and computer attitude: a model to predict the use of computerised information systems. AB - Computers are now essential technology in use by health workers. The literature shows that a number of factors effect the use of computers and many of them are related to attitude towards automation. Computer experience has been mostly used as a factor that effects computer attitude and its relationship with computer anxiety and computer use is not simple. Survey data from a study of 302 health workers, employed in a community setting was used to model the prediction of intention to use computers. The final model shows that 'positive computer experience' had a significant effect on computer attitude, computer anxiety and intention to use computers (both directly and indirectly). The model also confirms that those with a positive attitude towards computers had stronger intention to use computers. The value of these findings to health organisations in implementing automated systems is discussed. PMID- 10384589 TI - Toward clinical end-user computing: programmable order protocols for efficient human computer interaction. AB - We have developed and implemented an efficient method of managing routine patient care information as a programmable group order protocol. The purpose of protocol is to minimize a labor-intensive manual computer interaction by grouping clinically related routine orders as a single entity, thus to greatly speed up the time taken for manual entry such as keyboard stroke and/or mouse clicking. User programmability is added to facilitate insertion, deletion and update of order items to be a locally independent operation. A sequence of menu screen is also programmable when a change of standard operation is needed. Department specific order protocols are classified into four categories to improve user convenience. The degree of efficiency is measured by a number of key strokes and entry time. In most cases the time to enter order protocol with correction is found to take less than one minute with less than five key strokes. The method of order protocol entry clearly demonstrates end-user computing capability so that department specific requirements are resolved without resorting to computer department personnel. Flexibility of managing individual physician specific protocols is also beneficial enough to enhance the morale toward a hospital information system currently in use. PMID- 10384590 TI - Study of user preference of graphic user interface in laboratory information system. AB - In order to elucidate employees satisfaction levels in using graphic user interface. (GUI) in a laboratory information system (LIS), users attitudes toward GUI versus CUI. (Character User Interface) in a similar LIS were surveyed one month after implementing upgraded version of LIS in a tertiary care university hospital laboratory. The outcome of the study showed that approximately two third of users preferred to have GUI in LIS. There was no difference in preference of GUI of users whether they had previous experience with GUI or not. However the female and older employees tend to shy away from GUI. The employee productivity was improved with GUI although it takes slightly longer program loading time than that of CUI. In conclusion, the laboratory employee satisfaction was higher with GUI than CUI and their productivity was improved with GUI. PMID- 10384591 TI - Design of a 3D workbench interface for training in dental implantology. AB - The recent technological advances in the area of image processing and interactive computer graphics have lead to many sophisticated systems which support clinical experts in diagnosis and treatment planning tasks. In dentistry, the treatment with implants becomes more and more important. Although in the near future there will be increased demand for dentists who are trained in dental implantology, little effort has been made to introduce technologically new teaching methods into the curriculum of dentists. This paper describes the design of a virtual implant treatment planning system which is based on the concept of a three dimensional horizontal display. The system is utilizing different VR and multimedia techniques to achieve a highly interactive virtual workbench environment which is suited for class-room experimental education in dental implantology. PMID- 10384592 TI - On the way to a Web based hospital information system: concepts for the use of a medical data dictionary to present context sensitive information in an intranet environment. AB - Many authors have promoted the www-paradigm to build modern hospital information systems. However currently web-based applications are better suited to information "browsing" than to build complex data entry features. This prompted us to start with our first web based developments inside the Giessen University Hospital Information System in the field of pure presentation of stored knowledge and information. This article describes the concepts which will be used in Giessen to convert available information sources to the www paradigm and to implement context-sensitive knowledge presentation mechanisms inside the clinical information system. The approach is based upon a web-based medical data dictionary server. The data dictionary is used to map terms of interest, chosen from the clinical user during work with a HIS-application, to a semantic network of relationships. The dictionary server will follow those semantic links in order to find and display the webpages, which are linked to the subject. PMID- 10384593 TI - Happy birthday DIOGENE: a hospital information system born 20 years ago. AB - Since its birth in 1978, DIOGENE, the Hospital Information System of Geneva University Hospital has been constantly evolving, with a major change in 1995, when migrating from a centralized to an open distributed architecture. Since a few years, the hospital had to face health policy revolution with both economical constraints and opening of the healthcare network. The Hospital Information System DIOGENE plays a significant role by integrating four axes of knowledge medico-economical context for better understanding and influencing resources consumption the whole set of patient reports and documents (reports, encoded summaries, clinical findings, images, lab data, etc.) patient-dependent knowledge, in a vision integrating time and space external knowledge bases such as Medline (patient-independent knowledge) integration of these patient-dependent and -independent knowledges in a Case-Based Reasoning format, providing on the physician desktop all relevant information for helping him to take the most appropriate adequate decision. PMID- 10384594 TI - Migrating towards a client server architecture: a successful application in a 2,200-bed general hospital. AB - Asan Medical Center (AMC) completed a major migration process of the hospital information system from a mainframe towards an open Unix client server architecture from August of 1993 to August of 1996. Along with the east wing extension of AMC, the number of inpatient beds is greatly increased from 1,000 to 2,200 and information transaction increased from 300,000 to 700,000. A gradual departmental migration strategy with local area network connection and data conversion between the two systems were applied. The successful migration process towards a client server architecture provided improved user interface, enhanced flexibility and productivity of the system, better integration with diverse medical devices and improved networking flexibility. PMID- 10384595 TI - From legacy systems towards modern health information systems. AB - This article describes an approach of transferring legacy hospital information system to a modern HIS, which would support requirements of modern information age. There are several reasons, which force us to perform such a migration: from the weakness of design of legacy system to the need of covering new technologies (such as Inter/intranet) and new business conditions. The transition should be gradual, therefore a multilevel approach to the data is suggested, enabling us the transition, as well as including of the new, upcoming standards. The process of transition was started by the system covering the doctor's work. Responses to this part were very positive; therefore, we estimate that we are on the right way. We, however, know that the transitions will be neither fast nor cheap and this is the reason we shall have to find some more solutions. PMID- 10384596 TI - Towards semantic integration within an existing medical information system. AB - Talking about the problems of integration in medical information systems, the necessity to provide end users with a consistent and coherent view of patient's data, has been largely reported. In order to attempt this goal, systems need to perform semantic integration. We propose a pragmatic way to describe the semantics of the elements of a database, based on a bottom-up three steps process: 1. a back documentation of the elements of the system from their description contained in the data catalog of the database 2. a first semantic extension to transform a data catalog into a data dictionary 3. a second semantic extension to create a dictionary of the medical concepts from a data dictionary. This dictionary of concepts can be considered as the final result of "semantic integration". It contains a set of entities directly understandable by the end users. These entities are deduced or built from the elements collected and characterized in the data dictionary. This work reports the conceptual analysis and the implementation of such a data dictionary. PMID- 10384597 TI - A conceptual representation of clinical and managerial guidelines: the ATREUS workflow model. AB - In this paper we propose a workflow conceptual model able to represent clinical and managerial activities within healthcare structures, the ATREUS model. This model uses: a) a graphical representation which models the activities and the events that activate them; b) a textual representation of information related to: a set of conditions used for the control of activity execution; the actors who undertake the activity; the resources and tools necessary for its enactment, the clinical and managerial data generated by the activity execution; c) a state diagram which allows the control of the activity execution. The model allows modularity, activity nesting and temporal flexibility using a top-down refinement of processes. This model, unlike others, makes it possible to highlight the different types of decision involved in the performance of an activity. PMID- 10384598 TI - Vision of an 'automated' hospital information system. AB - This paper tries to describe a coherent vision of a possible next generation of HIS that rethinks how and what for computing is used in hospitals. Current systems, organized mainly around the 'database' and the 'communication' paradigm help the data processing to a great extent. At the same time they cannot be accepted as 'automated' systems, as handling of information is done mostly by end users, i.e. by human actors. Emerging methods enabling automated information handling are the following: integrated handling of different media, seamless communication among different systems, alternative input-output devices, tools for pro-active information handling. These technologies should be grouped to two main branches: technical advances in data handling and theoretical advances in knowledge handling. The advances in knowledge handling are really important: tools based on that can take over routine information handling tasks from human end users. To discuss automation in HIS it is useful to understand the process of information handling in general within the hospital. A suggested multidimensional information space, where information objects are gathered mainly along two axes, the 'patient axis' and the 'management' axis might be of help. Combinations of selected dimensions resulted in a space of an estimated 2,294,082 possible information handling situation types in an earlier publication. Automation of information handling tasks can be derived from this model. The authors suggest to automate certain tasks done usually by active actors of the information handling situation space. Software agents working 'on their own' are known entities in HIS systems. Two components are needed for automation: an organized data base where its content can be 'understood' and interpreted by an algorithm, with other words a knowledge base an algorithm, that covers a certain routine information handling task. Data bases of HIS should be re-thought in a way that enables automated processing to a greater extent. The development of data base technologies clearly point to this direction. If most of the data bases of a HIS will be like that, new generation of applications might be launched to use them. E.g. a software agent called 'patient assistant' could collect data from different sources and build a coherent and updated patient file. The results of a knowledge based, agent operated HIS should be the following: significantly less direct human involvement significantly less paper to be produced enhanced speed of data flow in general enhanced reliability by widespread watchdog functions PMID- 10384599 TI - "The Electronic Wardeni management of the data security access in a heterogeneous university hospital environment in Belgium. AB - A very flexible software system called "Electronic Warden" has been developed. It is based on a "client/server" architecture. It controls and manages the access right of the complex and heterogeneous data computer system at St Luc Hospital in Brussels. The electronic warden is independent of the other software applications of the hospital and is connected to them through API'S. The physical access is managed with the use of smart card and allows the electronic signature. The management of the users and their accesses to the data is run in a centralised or a decentralised way which allows a lot of flexibility. PMID- 10384600 TI - Hospital information system: reusability, designing, modelling, recommendations for implementing. AB - The aims of this paper are to precise some essential conditions for building reuse models for hospital information systems (HIS) and to present an application for hospital clinical laboratories. Reusability is a general trend in software, however reuse can involve a more or less part of design, classes, programs; consequently, a project involving reusability must be precisely defined. In the introduction it is seen trends in software, the stakes of reuse models for HIS and the special use case constituted with a HIS. The main three parts of this paper are: 1) Designing a reuse model (which objects are common to several information systems?) 2) A reuse model for hospital clinical laboratories (a genspec object model is presented for all laboratories: biochemistry, bacteriology, parasitology, pharmacology, ...) 3) Recommendations for generating plug-compatible software components (a reuse model can be implemented as a framework, concrete factors that increase reusability are presented). In conclusion reusability is a subtle exercise of which project must be previously and carefully defined. PMID- 10384601 TI - A practical object-oriented approach to a development of a next generation hospital information system. AB - KIND (stands for Kyushu university hospital Information Network Database) is a five years project, which aims to provide integrated services for patients, physicians, researchers and other hospital staffs. The final product of KIND is a next generation hospital information system. A physicians' clinical workstation, for example, integrated into a secured medical information network, can electronically develop a longitudinal medical record and interface with pharmacies, laboratories, medical specialists, and radiologists, as well as develop patient census and demographic profiles, in addition to doing electronic claims. Since clinical requirements on those medical records may vary for each case, we would like to have an essential data model under the hood. We decided to introduce domain analysis method to produce a relevant domain model. A domain analysis method captures the nature of business and helps us have an essential and extensible data model. Although there are several ways to describe a domain model, we chose an object-oriented description and consequently implementation using an object-oriented database system. Once we could have a decent domain model and implemented it as an object-oriented data model, application programs can utilize those data very easy without worrying extra efforts like finding complex queries including multiple joins. More over, if an application uses decent object-oriented technologies, it allows a user to access whole aspects of data transparently. This paper describes the architecture of KIND (the system) and outlines our domain model. In this paper, we also describe a practical application of several object-oriented technologies to develop a next generation hospital information system. PMID- 10384602 TI - A hospital information system based on Common Object Request Broker Architecture (CORBA) for exchanging distributed medical objects--an approach to future environment of sharing healthcare information. AB - Tightly related subsystems in a HIS have to exchange medical data flexibly by the data object rather than by the battery of the data. We developed a CPR subsystem based on Common Object Request Broker Architecture (CORBA) that retrieves and stores clinical information in the object-oriented database via Internet Intra ORB Protocol (IIOP). The system is hybridized with the legacy HIS applications on the client terminals. We believe that our solution and the experiences will contribute to the future CORBA-based environment in which computerized patient information is shared among hospitals, clinics, and tightly related systems. PMID- 10384603 TI - OOMM--Object-Oriented Matrix Modelling: an instrument for the integration of the Brasilia Regional Health Information System. AB - The development of Health Information Systems is widely determined by the establishment of the underlying information models. An Object-Oriented Matrix Model (OOMM) is described which target is to facilitate the integration of the overall health system. The model is based on information modules named micro databases that are structured in a three-dimensional network: planning, health structures and information systems. The modelling tool has been developed as a layer on top of a relational database system. A visual browser facilitates the development and maintenance of the information model. The modelling approach has been applied to the Brasilia University Hospital since 1991. The extension of the modelling approach to the Brasilia regional health system is considered. PMID- 10384604 TI - Why good hospitals get bad computing. AB - American Hospitals spend a lot of money for computing, but physicians and nurses are dissatisfied with the computing that they receive. No one is at fault. Rather, each person, acting in his or her best interest, unwittingly conspires to produce the unfavorable result. Until hospitals make major and fundamental changes in the way they purchase and manage computing, they will continue to spend large sums without commensurate return. PMID- 10384605 TI - The millennium bug: a prescription for action. AB - Healthcare delivery systems involving technology are vulnerable to year 2000 date problems. This paper describes the unique features of this problem as related to healthcare, proposes a classification to the problems, and structures an approach to begin resolving this looming disaster. PMID- 10384607 TI - An object-modeling method for hospital information systems. AB - Our goal is to increase the software productivity of hospital information systems (HISs) by applying object-oriented technologies. We propose a new object-modeling method for HISs that combines three general methods--an object modeling technique (OMT), class responsibilities collaboration (CRC) cards, and a use-case approach- to extract objects effectively. Our method also takes HIS functionality characteristics such as history and patient-staff linking into consideration to increase the reusability of objects. To test this method, we applied it to several HISs such as a drug-history system, a nursing-support system and an electronic patient record (EPR). We found that the software productivity of the drug-history system was more than doubled by using the combined method. Also, more than 40% of the logic classes could be reused in the nursing-support system and EPR. These findings confirmed that the proposed method can significantly increase the software productivity of HISs. PMID- 10384606 TI - Information system for a periodic examination and health promotion center. AB - The periodic examination and health promotion center (PEHPC) in Seoul National University Hospital has developed and adopted an information system supported by computers which use an electronic medical record to improve the quality of patient care, advance the science of medicine, lower health care costs, and enhance the education of health care professionals. This information system adopted the concept of incomplete and evolutionary systems to conduct the pursuit of practicalness and efficacy. It has increased efficiency to save costs and to enhance the quality of the medical service. It has also activated clinical research due to ease of managing data. We are also preparing for telecare. Telecare and WWW-using information system is postponed because the protection of a patients privacy is not established. PMID- 10384608 TI - An integrated environment for ECG processing. AB - The huge amount of information involved in clinical cardiological examination raises the need for efficient patient data management and for the fusion of modern information processing techniques in the everyday clinical workstation. An integrated medical information system has been developed in order to organize the local cardiological legacy system in the AHEPA hospital. An ODBC based database (like Ms SqlServer or MSAccess) holds patient and ECG data. The system incorporates data management (storing and retrieval) and data processing modules. The processing module is added as an independent DLL. The visualization component results in a better view of the information. The components are integrated in a friendly window interface that lets the doctor browse patient information, apply modern signal processing techniques, make special measurements and store them in the database for research reasons. PMID- 10384609 TI - Mobile PCIS: point-of-care information systems with portable terminals. AB - We have developed a Point-of-Care Information System (PCIS) that uses portable terminals and supports the entire loop of daily nursing work for the first time. Nursing work includes a great deal of indirect nursing work, such as documenting patient information, conferring with other health-care providers, and so on, as well as direct patient care. The time needed for this indirect work often exceeds the time actually spent directly caring for patients and needs to be reduced. Our system was developed with the goal of doing this to make nursing work more efficient. The system consists of personal digital assistants (PDAs) that nurses carry in the hospital wards and a server computer located in the nursing station. The system has three main functions: 1) Data Browsing provides patient information such as a brief history, vital-sign charts, and handwritten notes; 2) Schedule Planning helps nurses organize doctors' instructions and make To-do lists for the day; and 3) Care Management reminds nurses when they should execute the doctors' orders and provides tools for data entry. These three functions cover the entire loop of daily nursing work. The PDAs and the server computer can share up-to-date information by data transmission through telephone lines (low power cellular phones can be used), infrared devices, or RS-232C cables. Results from a preliminary hospital evaluation showed that use of the system can reduce the time needed for nursing documentation by 60% (a 10% saving of a nurse's total working time), and that more than 80% of the nurses who used the system felt it would make nursing work more efficient. PMID- 10384610 TI - The Erlangen university hospital communication hub--proprietary and standardised communication. AB - The University of Erlangen-Nuremberg contains 22 hospitals and 11 autonomous medical departments which are spread out over a large area in the city of Erlangen. The necessary connections of these units and their computer based subsystems to each other and to the medical computer centre via fibre optics cables is complete. The internal cabling of the individual units is largely completed. Based on this network the Erlangen communication hub allows medical subsystems of the Erlangen university hospitals to exchange data by two completely different methods. Since 1995 a communication data base, which is implemented using the relational data base system ADABAS D, contains data from the most important hospital systems. This data can be accessed by other medical systems. Thus the communication data base allows subsystems which do not have a standardised interface to implement proprietary system interconnections via access based on SQL. The capabilities of this interconnection are dependent on both the implementation and the data which is made available by the communication data base. This contains mainly basic patient data and the results of tests performed by various laboratory systems. In addition to this proprietary communication system we have since the end of 1996 a communication server which can also handle standardised message formats such as HL7, EDIFACT, DICOM3. Future subsystems which possess standard interfaces will be connected via this server. The connection of the patient management system IS-H and the central laboratory system to the database has been proceeding since the beginning of 1997. PMID- 10384611 TI - Plug and play--fiction or reality? AB - This paper discusses some experiences with the integration of a tumor documentation system into a distributed healthcare environment using a new European middleware technology. Although the middleware already offers a considerable support to the software engineer, further facilities have been suggested and partially implemented in order to accelerate integration scenarios. PMID- 10384612 TI - Application of a 3D display system for biplane cerebral angiography. AB - To improve the total safety for the biplane cerebral angiographic examination, we have developed a new three dimensional (3D) display system. By applying this system, shortening the examination time and reducing the risk of infection and embolization were achieved. We added just one graphic-workstation for 3D reconstruction to an ordinary biplane digital-cine-angiography system. This system can process the images of angiogram extempore in a short time and display the run of arterial vessels to the examiner. Biplane images were taken from the digital-cine-angiography system into the workstation through a video display. Three dimensional positions were calculated and displayed on a CRT monitor. It took from 15 to 45 seconds for the calculation. This system could display instantly the run of arterial vessels from any angle by using the computer mouse. Our system doesn't disturb the examination, but helps us to understand the run of the arterial vessels. This system was useful in cases where the arteries wound their way around or over-lapped each other. PMID- 10384613 TI - Detection of compact low-chromatin areas in cell nuclei images. AB - When analysing some DNA stained human cell nuclei using a light microscope or an quantitative image cytometer, compact low-chromatin areas (CLCA) can be observed. We are still not certain about the meaning and source of this phenomena. To enable the detection of CLCA by an image cytometer, a special image processing algorithm has to be developed and new nuclear cell features have to be designed. The presented image processing algorithm automatically detects CLCA in nuclei cell images taken from different tissues. The algorithm is composed of many basic image processing operations. The sequence of operations is determined by a priory knowledge about the properties of CLCA as a set of heuristic roles. The calculated CLCA features are CLCA area, perimeter, average intensity, compactnes and edge sharpness. The matching of the automatic CLCA detection and manual detection (performed by a biologist) was tested using 1400 cell nuclei. The results show a 83.4% match for the nuclei without CLCA and a 93.8% match for the cells with CLCA. PMID- 10384615 TI - Femoral anteversion: estimation by 3D modelling. AB - Femoral anteversion is the inclination of the femoral neck axis with reference to the tangent plane of the distal femur. For estimating femoral anteversion, following three major parameters are required; the neck axis, the long axis, the knee axis. Conventional methods on the basis of 2D images are ambiguous to determine these major axes. As the femur has a complex 3 dimensional structure, the 3 dimensional model should be applied for accurate and reliable measurement of femoral anteversion. In this paper, we model femur and define three parameters. The neck axis is defined from the femoral head and neck model. The long axis is determined from the cylindrical model of the femoral shaft. The knee axis is also determined from the model of femoral condyles. According to the definition of the femoral anteversion, the femoral anteversion is efficiently estimated from these models. 30 specimens were tested by conventional 2D imaging method, 3D imaging method which was developed by authors and the new 3D modelling method. The study provides accurate, fast and human factor free measurement for femoral anteversion. PMID- 10384614 TI - Development of videostrobokymography for the quantitative analysis of laryngeal vibratory pattern. AB - We developed a new analysis technique for the assessment of irregular vibratory movement of vocal folds. Successive frames of pre-recorded video images from videostroboscopy were transferred to computer memory and vibratory track of one selected point was described as a waveform by displaying the same lines of all frames along the vertical direction. Glottal area waveform, which was normalized with glottal length, was composed to quantitatively assess the overall vibratory pattern of vocal folds. By applying this technique, irregular vibratory patterns of multiple regions, such as asynchronized registration of glottal cycles, could be easily visualized. It would be possible to monitor and analyze the pathologic changes of vocal fold movement by means of this newly developed system. PMID- 10384616 TI - The Visible Human Project: a resource for anatomical visualization. AB - The National Library of Medicine (NLM) has long been a world leader in the archiving and distribution of the print-based images of biology and medicine. NLM has also been a pioneer in the use of computer systems to encode and distribute textual knowledge of the life sciences. NLM's Long Range Planning effort of 1985 86 foresaw a coming era where NLM's Bibliographic and factual database services would be complemented by libraries of digital images, distributed over high speed computer networks and by high capacity physical media. The NLM Planning Panel on Electronic Imaging recommended that NLM should undertake the building a digital image library consisting of computerized tomography (CT) and magnetic resonance (MR) images, and cryosection images of a representative, carefully selected and prepared male and female cadaver--the "Visible Human ProjectJ." The male and female Visible Human data sets are now being made available through a license agreement with the NLM. The data sets are supporting a wide range of educational, diagnostic, treatment planning, and commercial uses. The value of this international resource in the public domain increases through its application. Its utility will continue to grow as related databases are attached to it, and as more attributes are given to its image elements. PMID- 10384617 TI - Extending a teleradiology system by tools for 3D-visualization and volumetric analysis through a plug-in mechanism. AB - This paper describes ongoing research concerning interactive volume visualization coupled with tools for volumetric analysis. To establish an easy to use application, the 3D-visualization has been embedded in a state of the art teleradiology system, where additional functionality is often desired beyond basic image transfer and management. Major clinical requirements for deriving spatial measures are covered by the tools, in order to realize extended diagnosis support and therapy planning. Introducing the general plug-in mechanism this work exemplarily describes the useful extension of an approved application. Interactive visualization was achieved by a hybrid approach taking advantage of both the precise volume visualization based on the Heidelberg Raytracing Model and the graphics acceleration of modern workstations. Several tools for volumetric analysis extend the 3D-viewing. They offer 3D-pointing devices to select locations in the data volume, measure anatomical structures or control segmentation processes. A haptic interface provides a realistic perception while navigating within the 3D-reconstruction. The work is closely related to research work in the field of heart, liver and head surgery. In cooperation with our medical partners the development of tools as presented proceed the integration of image analysis into clinical routine. PMID- 10384618 TI - Development of multispectral autoradiography with solid state detector. AB - Autoradiography is a useful imaging technique to understand biological function by using radio isotopes (RIs). However, conventional autoradiography using X-ray film or Imaging Plate can not easily image distributions of different RIs simultaneously. Each tracers describes each biological functions, therefore we constructed here a multispectral autoradiography. The system consists of a solid state detector (SSD) with high energy-resolution to acquire multispectral information, a collimator with small aperture to restrict the region of observing area, and a translational stage to scan the specimen. The distribution of the RI of interest agent can be obtained by mapping spectrum intensity within the energy window to corresponding observed points. We confirmed the validity of this method by experiments with physical phantoms and tissues of rat's hearts. First, we imaged the phantom with 201Tl and 99mTc, and could specify the individual RIs. Next, from rectangular paper phantom soaked in 99mTc-labeled tracer, the spatial resolution of experimental system was estimated on about magnitude of x mm2. Finally, the sliced tissue of rat's heart was imaged. Here, we also propose a new imaging method to improve the spatial resolution and reduce data-acquisition time. It is based on reconstruction from projections like X-ray CT. PMID- 10384619 TI - Technical aspects of virtual liver resection planning. AB - Operability of a liver tumor is depending on its three dimensional relation to the intrahepatic vascular trees which define autonomously functioning liver (sub )segments. Precise operation planning is complicated by anatomic variability, distortion of the vascular trees by the tumor or preceding liver resections. Because of the missing possibility to track the deformation of the liver during the operation an integration of the resection planning system into an intra operative navigation system is not feasible. So the main task of an operation planning system in this domain is a quantifiable patient selection by exact prediction of post-operative liver function and a quantifiable resection proposal. The system quantifies the organ structures and resection volumes by means of absolute and relative values. It defines resection planes depending on security margins and the vascular trees and presents the data in visualized form as a 3D movie. The new 3D operation planning system offers quantifiable liver resection proposals based on individualized liver anatomy. The results are visualized in digital movies as well as in quantitative reports. PMID- 10384620 TI - Compression of color medical images in gastrointestinal endoscopy: a review. AB - This paper reviews the state of medical color digital imaging with respect to image compression. Only recently has the creation and storage of color medical images become technically and economically feasible. This has allowed medical images to become a part of an electronic medical record, to be used in telemedicine, or to be used for medical education. The Internet has become an important medium for the dissemination of medical images, whether through file transferring or through the World Wide Web. There is a growing need to evaluate the degree and types of image compression that are clinically acceptable (either for diagnostic or archival purposes) for different specialties. The focus will be on gastrointestinal endoscopic images, although work in other medical specialties will be mentioned. PMID- 10384621 TI - Design of a virtual reality laboratory for interdisciplinary medical application. AB - The Department of Medical Informatics of the University of Goettingen sets up a medical interdisciplinary Virtual Reality (VR) laboratory. The interdisciplinary approach for the design of the laboratory is based on a systematic, technical and application-orientated analysis. Its result led to the decision for a CAVE-like multi wall stereo projection (MWSP) system with networked workstation hardware. Within the boundary of an exemplary evaluation of the laboratory, its technical specifications and the validity in neuropsychological tests are supposed to be improved. Both techniques, Head Mounted Display (HMD) as well as multi wall stereo projection (MWSP) systems have a high degree of immersion. MWSP systems have a lower ratio of simulator sickness and a good visual fidelity. They can also be used as a multi-user environment. Networked workstations and high-end computers are compared in view of their costs and possible expansibility. PMID- 10384622 TI - Image analysis and pattern recognition for computer supported skin tumor diagnosis. AB - A new approach to computer supported recognition of melanoma and naevocytic naevi based on high resolution skin surface profiles is presented. Profiles are generated by sampling an area of 4 x 4 mm2 at a resolution of 125 sample points per mm with a laser profilometer at a vertical resolution of 0.1 micron. With image analysis algorithms Haralick's texture parameters, Fourier features and features based on fractal analysis are extracted. Genetic algorithms are employed successfully to select good feature subsets for the following classification process. As quality measure for feature subsets, the error rate of the nearest neighbor classifier estimated with the leaving-one-out method is used. Classification is performed with feed forward back-propagation network and the nearest neighbor classifier. Classification performance of the neural classifier is optimized using different topologies, learning parameters and pruning algorithms. The best neural classifier achieved an error rate of 4.5% and was found after network pruning. The best result with an error rate of 2.3% was obtained with the nearest neighbor classifier. PMID- 10384623 TI - Complete left ventricular wall motion estimation from cascaded MRI-SPAMM data. AB - We present a new paradigm which incorporates multiple sets of tagged MRI data (MRI-SPAMM) acquired in a cascaded fashion in order to estimate the full 3-D motion of the left ventricle (LV) during its entire cardiac cycle. Our technique is based on the extension of the volumetric physics-based deformable models, whose parameters are functions, which can capture the local shape variation of an object with a small number of intuitive parameters. By integrating a cascaded sequence of SPAMM data sets into our modeling technique, we have extended the capability of MRI-SPAMM and have provided an accurate representation of the LV motion from end-diastole to end-diastole to better understand cardiac mechanics. PMID- 10384624 TI - A marker-free system for the analysis of movement disabilities. AB - A major step toward improving the treatments of disabled persons may be achieved by using motion analysis equipment. We are developing such a system. It allows the analysis of plane human motion (e.g. gait) without using the tracking of markers. The system is composed of one fixed camera which acquires an image sequence of a human in motion. Then the treatment is divided into two steps: first, a large number of pixels belonging to the boundaries of the human body are extracted at each acquisition time. Secondly, a two-dimensional model of the human body, based on tapered superquadrics, is successively matched with the sets of pixels previously extracted; a specific fuzzy clustering process is used for this purpose. Moreover, an optical flow procedure gives a prediction of the model location at each acquisition time from its location at the previous time. Finally we present some results of this process applied to a leg in motion. PMID- 10384625 TI - Morphological classification of sperm heads using artificial neural networks. AB - In male reproducible health and fertility and IVF (in-vitro fertilization), morphological analysis of sperm has been most important. But the traditional tools for semen analysis are subjective, imprecise, inaccurate, difficult to standardize, and reproduce mainly due to their manually oriented operations. The purpose of morphological analysis of sperm is to microscopically type-classify sperm according to their morphological characteristics of heads. Until now, the strict criteria method has long been used in clinic to discriminate normal sperm from abnormal. This method cannot classify the diverse groups of abnormal sperm in detail and shows large variations in inter-operators and intra-operator. In this paper, we have studied a new method of sperm morphological classification using artificial neural networks that are widely used in pattern recognition and image processing. With a multi-layer perceptron trained by the error back propagation algorithm, profile features from digitized sperm images were classified into four classes that consisted of one normal group and three abnormal groups according to their morphological characteristics. PMID- 10384626 TI - A hospital-wide distributed PACS based on intranet. AB - A hospital-wide Picture Archiving and Communication System (PACS) is currently under development at the University Hospital of Geneva. After a first implementation including two oneterabyte optical libraries, the system is expanded to integrate all the imaging modalities of the hospital. The new storage requirement is 10 terabytes to cover three year archive. A large distributed image archive has been designed including new archive servers for long-term storage and display servers for medium-term storage. The acquisition, archive and distribution cycles are performed using separated networks combining Fast Ethernet and Ethernet. Image files are distributed to the wide-hospital using a prefetching strategy or an Intranet server, RADIOLAB. The first mode takes advantage of the fully integrated hospital information system DIOGENE 2 to allow the automatic retrieval of studies in advance. The second mode provides a convivial study selection from any conventional WWW (World Wide Web) browser. Image files are then transmitted to the user's display station using HTTP (HyperText Transfer Protocol) and handled by OSIRIS software, which acts as a helper or viewer. Such a system is expected to meet the time requirement, which is less than three seconds per image. PMID- 10384627 TI - Modeling and performance analysis of image transfer in PACS. AB - Performance simulation plays an important role in the design of picture archiving and communication systems (PACS). This paper presents simulation models of a PACS in a context of outpatient clinical workflow. Models, which are discussed focusing on shared facilities are resources of the system, consist of two levels. One is the workflow level and the other is the physical level. Patients medical staffs and images are picked up as the important system resources in the workflow level. A network, hard disks and network interface buffers of PACS components are mainly described in the physical level. Models are described with Petri Nets in both levels. Two examples of performance evaluation are also presented. Both of them estimate transfer time of computed radiology images for outpatient consultation. In the first example, both the data transfer rate of hard disks and network speed are varied under fixed system loads. While in the second example, system loads are varied against given system components. From the simulation results, following points are summarized. If physicians want to get a set of images in 5 seconds, both high performance hard disks or disk arrays and high speed network such as 100 Mbps networks are required. And it is effective for high speed network to be used in the acquisition phase. If we have to use the Ethernet and ordinary workstations, the PACS should be used in a single clinic. In conclusion this modeling method makes models simple and is quite useful for the performance evaluation of PACS in the early design phase. PMID- 10384628 TI - Three-tiered integration of PACS and HIS toward next generation total hospital information system. AB - The Seoul National University Hospital (SNUH) started a project to innovate the hospital information facilities. This project includes installation of high speed hospital network, development of new HIS, OCS (order communication system), RIS and PACS. This project aims at the implementation of the first total hospital information system by seamlessly integrating these systems together. To achieve this goal, we took three-tiered systems integration approach: network level, database level, and workstation level integration. There are 3 loops of networks in SNUH: proprietary star network for host computer based HIS, Ethernet based hospital LAN for OCS and RIS, and ATM based network for PACS. They are linked together at the backbone level to allow high speed communication between these systems. We have developed special communication modules for each system that allow data interchange between different databases and computer platforms. We have also developed an integrated workstation in which both the OCS and PACS application programs run on a single computer in an integrated manner allowing the clinical users to access and display radiological images as well as textual clinical information within a single user environment. A study is in progress toward a total hospital information system in SNUH by seamlessly integrating the main hospital information resources such as HIS, OCS, and PACS. With the three tiered systems integration approach, we could successfully integrate the systems from the network level to the user application level. PMID- 10384629 TI - Implementation of a low-cost PACS/CR for clinical use in Yonsei Cardiovascular Center. AB - The PACS/CR for clinical use in Yonsei Cardiovascular Center has been designed and implemented. Our system is open architecture to comply with emerging standards such as DICOM. database SQL, TCP/IP and to reduce operational and maintenance costs, PC-based low cost workstations running Microsoft Windows, database as Microsoft SQL based on Client/Server, Long-term storage using CD-ROM Jukebox are developed. Also, auto routing and image pre-fetching are implemented. PMID- 10384630 TI - Experimental assessment of the feasibility of integrating an endoscopic imaging system into an existing hospital information system. AB - The National Cheng Kung University (NCKU) Hospital operates a comprehensive integrated hospital information system. This was designed and installed as an integral component of the building of the hospital, which opened in 1988. The information system provides a service to all staff in the hospital, administrative and clinical, across the whole of the hospital. To make sure that the information is accessible, where and when it is needed, a comprehensive communications network consisting of both hardware and software mechanisms supports the information system. This paper explores the requirements for a computerised image system for endoscopy and assesses two approaches to the implementation of such a system, as a stand-alone system and as a subsystem of the NCKU hospital information system already in place. The latter would satisfy the original design specification of developing a single system to cover all aspects of hospital operation but places additional demands on the endoscopy systems designer to ensure integration. Both operational modes are set out in the paper and their implications assessed. PMID- 10384631 TI - An architecture for implementing customizable medical image processing systems. AB - Monolithic image processing systems containing a superset of imaging algorithms are difficult to use and require specialized knowledge of image processing. Thus they increase the workload of medical personnel instead of making the work situation easier. Customizable medical image processing systems on the other hand may be easily adapted to address various problems in the medical image processing domain integrating only the necessary subset of image processing functionality presented in on intuitive way. In this work we present an architecture for creating customizable image processing systems for the medical domain. We address three major topics: 1.) easy, goal-oriented customization of imaging systems by using a generalized algorithm model and repository, 2.) dynamic, data-oriented parameterization of the selected algorithms and 3.) semi-automated generation of user interface components for each new algorithm to be inserted in an imaging system based on cognitive ergonomics. We conclude with the presentation of an initial implementation of the architecture in form of an object-oriented framework for the creation of components for customizable medical imaging systems. PMID- 10384632 TI - Web based 3-D medical image visualization on the PC. AB - With the recent advance of Web and its associated technologies, information sharing on distribute computing environments has gained a great amount of attention from many researchers in many application areas, such as medicine, engineering, and business. One basic requirement of distributed medical consultation systems is that geographically dispersed, disparate participants are allowed to exchange information readily with each other. Such software also needs to be supported on a broad range of computer platforms to increase the softwares accessibility. In this paper, the development of world-wide-web based medical consultation system for radiology imaging is addressed to provide platform independence and greater accessibility. The system supports sharing of 3 dimensional objects. We use VRML (Virtual Reality Modeling Language), which is the defacto standard in 3-D modeling on the Web. 3-D objects are reconstructed from CT or MRI volume data using a VRML format, which can be viewed and manipulated easily in Web-browsers with a VRML plug-in. A Marching cubes method is used in the transformation of scanned volume data sets to polygonal surfaces of VRML. A decimation algorithm is adopted to reduce the number of meshes in the resulting VRML file. 3-D volume data are often very large in size, hence loading the data on PC level computers requires a significant reduction of the size of the data, while minimizing the loss of the original shape information. This is also important to decrease network delays. A prototype system has been implemented (http://cybernet5.snu.ac.kr/-cyber/mrivrml .html), and several sessions of experiments are carried out. PMID- 10384633 TI - Results of European projects improving security of distributed health information systems. AB - The challenge for improvement of quality and efficiency of health care systems causes the development and promotion of "Shared Care" in all developed countries. Distribution, decentralisation, and specialisation of health care must be joint with an extended communication and co-operation between the different care providers. Fulfilling the shared care paradigm, care supporting health information systems has to be distributed, interoperable, and scaleable too. Communication and co-operation across organisational, regional, and even national boundaries is bearing high threats and risks regarding security and privacy of medical and personal information of both patients and health professionals. Involved in several security projects funded by the European Union, the Medical Informatics Department and the regional Clinical Cancer Registry at the University of Magdeburg are piloting a secure regional distributed medical record system for cancer diseases. Requirements, solutions, and experiences are presented and discussed. PMID- 10384634 TI - Three computational systems for disclosing medical data in the year 1999. AB - Today most organizations release and receive medical data with all explicit identifiers, such as name, address, and phone number, removed in the incorrect belief that patient confidentiality is maintained because the resulting data look anonymous. We examine three computer programs that do maintain patient confidentiality when disclosing electronic medical records: the Scrub System which locates personally-identifying information in letters between doctors and notes written by clinicians; the Datafly System which generalizes data within the record based on a profile of the recipient at the time of access; and, the mu Argus System which is becoming a European standard for disclosing public use data. The techniques presented in these systems help protect confidentiality in the face of a changing globally-networked society with immediate access to volumes of personal data. PMID- 10384635 TI - PCASSO: a secure architecture for access to clinical data via the Internet. AB - Patient Centered Access to Secure Systems Online (PCASSO) is a National Information Infrastructure research project funded by the US National Library of Medicine (NLM). PCASSO is specifically designed to address the problem of enabling secure access to health information over the Internet, not just for healthcare providers and medical researchers, but also for patients to view their own medical records. The project is using familiar Web technologies to support the search and retrieval of information, including patient demographics, medications, lab tests, and transcription reports. State-of-the-art security technologies protect patient privacy and the integrity of patient information. This paper describes the security architecture of the PCASSO system. PMID- 10384636 TI - Security aspects of medical file regrouping for the epidemiological follow-up. AB - To carry out epidemiological studies at a regional level, one may need to link information collected by medical doctors working either in hospitals or in private offices or laboratories. The first problem is to respect the European legislation on nominal data processing, which does not allow the linkage of nominal files. As a consequence, we have developed an anonymous record linkage procedure, which ensures an irreversible transformation of identity and allows the linkage of rendered anonymous files. The second problem is to ensure data security during the transmission and we discuss the advantages of different methods of communication such as the norms X400 and Internet protocols. PMID- 10384637 TI - Methods of responding to healthcare security incidents. AB - This paper considers the increasing requirement for security in healthcare IT systems and, in particular, identifies the need for appropriate means by which healthcare establishments (HCEs) may respond to incidents. The main discussion focuses upon two significant initiatives that have been established in order to improve understanding and awareness of healthcare security issues. The first is the establishment of a dedicated Incident Reporting Scheme (IRS) for HCEs, enabling the level and types of security incidents faced within the healthcare community to be monitored and advice appropriately targeted. The second aspect presents a description of healthcare security World Wide Web service, which provides a comprehensive source of advice and guidance for establishments when trying to address and prevent IT security breaches. The discussion is based upon work that is currently being undertaken with the ISHTAR (Implementing Secure Healthcare Telematics Applications in Europe) project, as part of the Telematics Applications for Health programme of the European Commission. PMID- 10384639 TI - Confidentiality principles for medical records on the World Wide Web. AB - We have proposed elsewhere a strategy for releasing medical records via the World Wide Web. The philosophical underpinnings of this proposal balanced a need for access with a need for confidentiality of medical information. Other balance points could have been chosen, and methods of stronger and weaker protection of confidentiality are presented here along with the rationale behind the selected strategy. PMID- 10384638 TI - Privacy and efficiency in patients focused health care processes. AB - Today's health care industry is striving to achieve the development and deployment of computer-based patient records for improvements in health care quality, cost, and access. This rapidly increasing use of computer techniques in the field of medicine and health care has created an urgent need for the dissemination of information on data protection. We present in this paper a workflow-based approach to ensure privacy and efficiency in clinical information systems and discuss the presented solution according to its practicability in daily clinical use. PMID- 10384640 TI - Access control and system audit based on "patient-doctor relation and clinical situation" model. AB - Both confidentiality of privacy and the data sharing between healthcare practitioners are required in hospital information systems. A new access control method has been designed by the "patient-doctor relation and clinical situation at the point-of-care" model in addition to the traditional "account and password" mechanism. This method can; (I) allow flexible data access in need, (2) afford accurate access audit, (3) suppress inappropriate access. PMID- 10384641 TI - The Health On the Net Code of Conduct for medical and health web sites. AB - The growth in the number of medical and health web sites and the varying quality of medical and health information currently available on the World-Wide Web has created the need for guidelines to help homogenise this quality. Health On the Net Foundation (HON) has initiated the Code of Conduct (HON-code) in response to concerns expressed to the Foundation, by members of the Net community. This article presents the HON-code's principles since its launch in 1996 and its status in 1997. This initiative offers information providers good practice guidelines and offers users an indication of commitment to quality by those providers. PMID- 10384642 TI - The information needs of health agency board members: a health information system challenge. AB - Most hospitals and health agencies in developed countries, whether in the private or public sector, have 'advisory committees' or 'boards of management'. Members of these bodies have significant responsibilities for the management of their organisations, but usually serve on these bodies on a part-time and voluntary basis. They may or may not have specific expertise in health care or management but are often selected because they are representative of parts of the community, are members of relevant organisations (medical bodies or, perhaps, staff unions) or belong to the correct political party. In previous years the amount of information about the health care agency and health systems in general which was given to the board was usually closely controlled by the senior staff of the agency. While the information often emphasised financial statements or gross usage figures, it seldom included projections of future trends or comparisons with information for other like agencies and was frequently very out-dated by the time that it was assembled. For discussion purposes in this paper all individuals who serve on advisory or management committees as well as members of legally constituted Boards of Directors will be referred to as 'board members'. The responsibilities of these board members differ from the management responsibilities of CEOs and other senior executives, but those involved do require access to some management information if they are to adequately discharge their responsibilities. The role of such 'boards' or 'authorities' is similar to that of a board of trustees or directors in industry with many of the same responsibilities, except that healthcare agencies are 'people' agencies, not producers of 'widgets'. This paper will outline some of the information needs of board members and discuss how these needs may be addressed as part of a total management and health information system. PMID- 10384643 TI - Healthcare systems process reengineering for developing countries: a report to IMIA Working Group 9. AB - To discover what kinds of health informatics the developing countries need, we look first to the underlying social structures that informatics is to support. Developing countries face severe problems in their goal of assuring access to quality healthcare to their citizens. Nor is it comforting to look to the example of the developed countries, themselves uncomfortable with the tremendous commitment of resources that they use. It seems clear that if developing countries are to succeed their systems for assuring health will need to achieve radical improvements over those currently employed by the developed countries. They need to build systems that are directed to the heart of the problems; they need to make the best use of their existing human and economic resources; and they need to do this while respecting the human potential and aspirations of their people. Any such system will need to be built around planning, organizing and measuring the long-term, rather than episodic, health of citizens. In particular, health interventions need to be moved upstream where they are more effective and less expensive. The job of health informatics is to provide the fundamental enabling infrastructure that would allow individuals to cooperate in the fluid, effective ways required. We describe some of the requisites for such a system. Certainly, its construction will depend on the use of an open, standards based distributed object software architecture. PMID- 10384644 TI - Medical informatics in the new millennium. AB - In a period of social transformation, we must reinvent health care. For guidance, we can look to the evolving discipline of medical informatics and to the patterns of investment in the practice arena. A top ranked application need, the computerized patient record (CPR) offers cost savings and supports clinical quality and ambulatory care. In the new millennium, we need to define our values with precision and use technology to achieve quality health care. PMID- 10384645 TI - Health informatics and the humanities. AB - Researchers and professionals from the Humanities are new within the science of health informatics, where researchers and professionals from the natural sciences have so far de-fined the problems and their solutions. A cross-disciplinary collaboration can be difficult due to factors such as status and competition, to insufficient co-operation qualifications, and due to different definitions of problems. This paper presents examples of differently formulated health informatics problems. Cross-disciplinary collaboration is necessary but collaboration demands openness, curiosity, and readiness from researchers and professionals as well as it demands know-edge of the processes in cross disciplinary cooperation. Considerations for cross-disciplines were actualized with the foundation in Denmark of the cross-disciplinary institution: The Virtual Centre for Health Informatics. PMID- 10384646 TI - Health or uni-disciplinary informatics initiatives--why are we where we are today? AB - This paper looks at the development of health informatics from a segregated uni disciplinary beginning to an environment in which multi-disciplinary health informatics and uni-disciplinary work co-exist reasonably harmoniously. However, the paper considers some of the catalysts and inhibitors which have made the transition not as smooth or wide ranging as the technological developments could have supported. Organisational and cultural factors appear to have played a part in the migration as it has emerged. Identification and understanding of the key issues may help in progressing health informatics towards a mature state. PMID- 10384647 TI - Setting a national research agenda in nursing informatics. AB - An active program of research assures the development and evaluation of nursing informatics solutions to the challenges of contemporary patient care. Experts in nursing informatics research participated in a two-part electronic mail survey of research priorities. Priorities identified included formalization of nursing vocabularies, design and management of databases for nursing information, development of technologies to support nursing practice, use of telecommunications technology in nursing, patient use of information technology, identification of nurses' information needs, and systems modeling and evaluation. Many of these priorities are similar to those advanced in the 1993 US PHS NINR PEP Report on Nursing Informatics. Additionally, the findings suggest the need for greater emphasis on the application of emerging technology to nursing practice problems, and the expansion to consider patients as direct users of information systems. PMID- 10384648 TI - Organizational & medical ontologies for co-operative patient management. AB - A new research paradigm is emerging based on the Multi-Agent System (MAS) architectural framework, allowing human and software agents to interoperate and thus cooperate within common application areas. In this paper, we discuss an ontological foundation for a prototypical health care MAS, that is, a so-called Distributed Healthcare Information System (D-HIS), viewed as an organization of cognitive agents and making use of an ontological library written in the standard language Ontolingua. PMID- 10384649 TI - People and organizational aspects of medical informatics. AB - Positive outcomes come to health care organizations that are doing the right things well, that is, their organizational strategies are aligned with their environments and they are executing those strategies well. Likewise, the informatics strategies must also be aligned with the organization's strategies. Without this congruence, informatics does not have the potential to have a substantial positive impact on the overall organizational outcomes. The execution of the informatics strategies must also be exemplary; however, the change processes required for achieving the desired informatics and organizational outcome goals are demanding and complicated. Implementing them in extremely complex organizations that operate on a 7-day by 24 hour basis is not easy. However, we are constantly learning more about complex change processes and the ways we can better manage them to improve our needed informatics outcomes. The challenge is to build upon the existing research base to more us even further ahead. PMID- 10384650 TI - An examination of the variables associated with the adoption of a computerized innovation (an operating room simulation model). AB - The purpose of this study was to describe the variables that influenced senior staff to decide to adopt (or reject) a computerized innovation (Operating Room Simulation Model) and to describe the decision process at four hospitals. Rogers' Innovation-Decision Process Model (1983) and Model of the Innovation Process in Organizations (1983) formed the conceptual framework for this study. Five specific variables (relevant advantage, compatibility, complexity, trialability, observability), that were shown in the literature to increase the probability of an innovation to be adopted, were investigated. The results of this study indicated that there was a match between relevant advantage and adoption and no match between compatibility and adoption. There were insufficient data to determine if a match existed between complexity, trialability and observability with adoption of the Model. The links between the organizational variables (agenda setting, matching, decision) and adoption of the OR Simulation Model were not conclusive. PMID- 10384651 TI - The need for a holistic view of telemedicine, focused on patients and society as prime stakeholders. AB - Telemedicine provides the scope to remove the barrier of distance from delivery of health care. In so doing it has the potential to strengthen primary care and local health delivery, and to optimise the use of secondary and tertiary specialist resources. However, it can have disbenefits and perverse effects which are as yet inadequately addressed. Development of telemedicine to date has been led by new technological opportunities, linked to clinical interests. Whilst not inappropriate, this alone is not adequate. All health care activities should be to promote health and treat ill-health, and therefore the interests of the consumer and of society should predominate. More complete research and policy analysis of telemedicine are therefore needed. This paper argues for a more holistic and integrated view of telemedicine to be taken, to ensure that technology is harnessed to meet need. To this end, it puts telemedicine into the overall context of health care interests, and describes methodologies to assess patient attitudes and societal priorities. Given the need to avoid the unanticipated adverse effects which have bedevilled other new health care technologies, and the particular risks of uncontrolled consultations and commerce across national boundaries, the paper goes on to advocate routine use of integrated risk analysis of telemedicine applications at research and policy levels. The need for an effects-orientated analytic framework, and for international policy collaboration to protect users of services available by global telecommunications, is identified. PMID- 10384652 TI - Medical affective computing: medical informatics meets affective computing. AB - "The need to cope with a changing and partly unpredictable world makes it very likely that any intelligent system with multiple motives and limited powers will have emotions." [1] From advisory systems that understand emotional attitudes toward medical outcomes, to wearable computers that compensate for communication disability, to computer simulations of emotions and their disorders, the research agendas of medical informatics and affective computing--how and why to create computers that detect, convey, and even have emotions--increasingly overlap. Some psychiatric and neurological researchers state their theories in terms of actual or hypothetical computer programs. Adaptive intelligent systems will increasingly rely on emotions to compensate for their own conflicting goals and limited resources--emotional reactions about which psychiatrists and neurologists have special insights. DEP2 (Depression Emulation Program 2) is a computer simulation of adaptive depression--learning from explainable patterns of failure in autobiographical memory--that simulates many depressive behaviors. In the terminology of fault-tolerant computing, adaptive depression involves fault detection (triggered by failure), fault location (strategic retreat and failure diagnosis), and fault recovery (return to on-line operation). DEP2 relies on subsystems whose structures and behaviors are based on popular hypotheses about left and right brain hemispheric function during depression and emotion. DEP2 and its predecessors, DEP and DEPlanner, are relevant to psychiatric and neurological informatics, and to the design of adaptive autonomous robots and software agents. PMID- 10384653 TI - Global healthcare and the flux of technology. AB - We begin with the inescapable observation that healthcare informatics is merely one of the many endeavors that is following a turbulent but nearly inescapable path into a digital future. Our objective in this paper is to describe as best we can the overall geography of the general path we appear to be on, to anticipate some of our future checkpoints along the way, to identify some of the roughest transitional passages, and to offer this as one guide among many to those who have volunteered to do the steering into this exciting, electronic unknown. PMID- 10384654 TI - The ethical imperative of child health population-based data a top-down/under-up analysis from the United Kingdom and Australia. AB - The policy move in both the United Kingdom and Australia to competitive primary care child health services (including preventive services and surveillance) has many advantages, but without secure and epidemiological information systems has the potential to put children at risk. More fundamentally, it produces the perverse effect of operating directly in opposition to the desirable policies of audit, outcomes studies, and evidence-based health care. Moreover, generations of children stand to be disadvantaged if environmental and ecological risk factors, as well as adverse clinical outcomes, cannot be identified quickly. Health informatics, linked to professional duties of clinical and fiduciary accountability, provide a way out of this conundrum. Standard clinical data sets, linked to invoicing procedures, would enable the development of summary records which could underpin integrated surveillance whilst also fuelling the new public health. Appropriate professional supervision, confidentiality and security measures, and agreement of data sets, would be necessary but not difficult in informatics terms. As health policy and delivery move forward, it is important not to lose the best aspects of previous systems and opportunities of future technologies. PMID- 10384655 TI - Towards an ethical protocol in mental health informatics. AB - Application of informatics to mental health has the potential to benefit a disadvantaged yet important patient group. At the same time, difficult issues are raised with regard to data protection and ethics because many patients are not able to express informed and rational views, and this increases their vulnerability. However, developments in this field are limited, and there is little literature. Generic data protection legislation and guidance assumes that data subjects are mentally competent adults. This paper distils key ethical issues and principles in mental health informatics, and describes a process which has initiated the identification of "reasonable" practice. PMID- 10384656 TI - Health informatics in Canada: a vision for the next millennium. AB - Around the world, informatics has been cited as a key enabler of health sector reform. Recent reform programs in Canada, reflecting this global consensus, have emphasized the importance of quality information and information technology in achieving their goals. This paper describes the current status of health informatics in Canada and provides a vision for the evolution of Canada's health information infrastructure. PMID- 10384657 TI - Health care workers and their needs: the forgotten shadow of AIM research. AB - The field of AI in Medicine (AIM) seems to have accepted that decision support is, and will be, needed within most medical domains. As society calls for cost effectiveness, and human expertise or expert guidance are not always available, decision support systems (DSSs) are proposed as the solutions. These solutions, however, do not necessarily correspond with the basic needs of their targeted users. We will show this through a review of the literature related to health care workers and the various factors that have an influence on their performances. Furthermore, we will use these empirical findings to argue that the AIM community must go beyond its decision support philosophy, whereby the gaps in human expertise are filled in by the computer. In the future, joint emphasis must be placed on decision support and the promotion towards independent and self sufficient problem solving. In order to implement this paradigm change, the AIM community will have to incorporate findings from the research discipline of AI in Education. PMID- 10384659 TI - A signal analysis method for retinal image velocity computation of vestibular ocular reflex eye movement investigations. AB - The vestibulo-ocular reflex preserves clear vision when the subject's head is moving rapidly. These eye movements are measured in order to investigate equilibrium diseases and disorders in otoneurology. The retinal image velocity is calculated to be a difference signal between the head movement velocity and the eye movement velocity. The eye movement position signal as well as the head movement position signal of the subject are concurrently measured when the subject is asked to move his head according to a stimulus and his gaze is fixated to a target. Eye movements are electro-oculographically recorded and head movements with a potentiometer. Tests performed to both patients of an equilibrium laboratory and normals exemplify that the method is applicable to indicate some otoneurological diseases. PMID- 10384658 TI - Establishing the feasibility of cross-mapping independent child health data sets as a means of comparing health status and service quality. AB - Benefits of comparative data analysis for quality assurance and policy development are clear, but are difficult to establish through traditional means of prospective projects. This paper describes a pilot project to test an alternative method, namely seeking to identify areas of commonality amongst very different pre-existing data sets to provide a starting point. Advantages and disadvantages of this method, together with the critical factors identified in the pilot study, are described. Whilst developed in child health, the lessons learned are relevant for any other client groups. PMID- 10384660 TI - Multimodal time-variant signal analysis of neonatal EEG burst patterns. AB - It can be shown that dominant rhythmic signal components of neonatal EEG burst patterns (discontinuous EEG in quiet sleep) are characterized by a quadratic phase coupling (bispectral analysis), i.e. a multiplicative interaction (connection) between the underlying electrophysiological processes can be assumed. By means of pattern recognition algorithms as well as time-variant spectral and coherence analysis, a so-called "initial wave" (narrow band rhythm within a frequency range of 3-12 Hz) can be demonstrated within the first part of the burst pattern. The detection of this signal component and of the quadratic phase coupling is more successful in the frontal region. By means of amplitude demodulation of the "initial wave" the phase coupling can be attributed to an amplitude modulation. The results were derived from 6 neonates (20 burst patterns for each neonate; 8-channel recordings). A 16-channel EEG-recording was analyzed for one neonate. PMID- 10384661 TI - Detection of epileptiform activities in the EEG using neural network and expert system. AB - This paper proposes a multichannel spike detection in long term EEG monitoring for epilepsy. It is achieved by wavelet transform(WT), artificial neural network(ANN) and the expert system. First, a small set of wavelet coefficients is used to represent the characteristics of a single channel epileptic spikes and normal activities. The purpose of this WT is to reduce the number of inputs to the ANN. Next, three layer feed-forward network employing the error back propagation algorithm is trained and tested using parameters obtained by the WT. Spikes are identified in individual EEG channels by 16 identical neural networks. Finally, 16-channel expert system based on the context information of adjacent channels is introduced to reject artifacts and produce reliable results. In this study, epileptic spikes and normal activities were selected from 32 patient's EEGs (the seizure disorder: 12, normal: 20) in consensus among experts. The result shows that the WT reduced data input size and the preprocessed ANN had 97% sensitivity and 89.5% selectivity, which were more accurate than that of ANN with the same input size of raw data. In clinical result, our expert rule system, which uses neighboring channel informations, was capable of rejecting artifacts commonly found in EEG recordings. PMID- 10384663 TI - Quality management in the doctor's consulting room. AB - Clinical research and the measurement of quality in healthcare have similar properties. Clinical research is generally restricted to university and teaching hospitals because of it's time consuming nature while quality management and subsequent quality measurement are of interest to all medical doctors. To serve both purposes a Kaplan Meier satellite module was developed as an add-on to a medical workstation. Kaplan Meier survival statistics has interesting possibilities as a clinical tool for the measurement and graphic visualisation of quality of care. It does express the clinical behaviour of a patient population in terms of cumulative survival, patency or any other binary phenomenon. A vascular surgeon collected cumulative patency data on 340 vascular reconstructions with this Kaplan Meier module within 6 months without any perceptable influence on the normal outpatient clinic routines. The module is easy to operate, easy to maintain and very adaptable to any clinical question. PMID- 10384662 TI - A model for enhancing worldwide personal health and wellness. AB - Individuals must increasingly take control of managing their own health affairs. This requires access to quality information that is not easily obtained from traditional social institutions. NetWellness is an electronic consumer health information service that provides a model for reaching the goal of enhancing personal health and quality of life. [1] We present a vision of consumer health information delivery in the 21st century, and a model for reaching that vision. Our experience to date, progressing through a five-phase model, is aimed at providing the best health information possible to the widest population possible. PMID- 10384664 TI - Components of the optimal ambulatory care computing environment. AB - We present here a framework of core components of an ambulatory care computing environment, based on clinical and functional needs and workflow scenarios. We have established this framework through the use of two study devices: a vision of the clinical office of the future, and a survey of possible computer applications, both designed to help clinicians and practice directors communicate their information needs to systems designers. Clinicians prioritize applications based on strategic and practice goals: support for clinical users' workflow, improved quality of care, reduced cost of care, and the ability to measure performance and status. By reorganizing the needed functionality from a clinical viewpoint into a technical viewpoint, we are able to identify core information components for systems design. Based on this analysis, information needs in the ambulatory environment can be divided into five primary functions: patient data retrieval, documentation, communication, knowledge resources, and aggregate reporting. Three other fundamental processes--knowledge-based interventions, information integration, and confidentiality--run through all of these front-line functions. Component applications and data structures built with this framework in mind will afford a maximum combination of functionality and flexibility to handle future changes in the clinical environment. PMID- 10384665 TI - Virtual patients for a virtual hospital. AB - With the introduction of VBscript & Active-X it is now possible to construct interactive websites with ease. This paper discusses the author's experience in setting up TWO different 'VIRTUAL PATIENT' websites. One such site, named VIRTUAL PATIENT 97, is based upon a goal based scenario, in this case the clinical management of common clinical problems like upper gastro-intestinal bleeding in a surgical ward. The second website, CLICK 'n' LEARN, is essentially a health education website providing material on common chronic illnesses like diabetes, hypertension and asthma. The first module deals with aspects of self care in diabetes. Included are textual as well as video clips which show various elements of diabetes self care: like the administration of insulin or the usage of a glucometer. An added twist is a target oriented interface borrowed from the TAMAGOTCHI. If the user dutifully looks after his virtual diabetic, it will thrive & the disease will be well controlled, else the virtual patient deteriorates in accelerated time. This allows for safe experimentation & promotes awareness of the importance of self care in the total management of chronic illnesses. PMID- 10384666 TI - Telemedicine and medical informatics in the multimedia super corridor: the Malaysian vision. AB - The practice of medicine, with its wide range of environmental conditions and complex dependencies, has long been used as a test bed for various advanced technologies. Telemedicine, as conceptualised within the Multimedia Super Corridor (MSC) context, is seen as the application of several relatively mature technologiesartificial intelligence (AI), multimedia communication and information systems (IS) amongst othersso as to benefit a large cross-section of the Malaysian population. We will discuss in general terms the Malaysian vision on the comprehensive MSC telemedicine solution, its functionality and associated operational conditions. In particular, this paper focuses on the conceptualisation of one key telemedical component i.e. the Lifetime Health Plan (LHP) system, which is eventually intended to be a distributed multi-module application for the periodic monitoring and generation of health-care advisories for upwards of 20 million Malaysians. PMID- 10384667 TI - Information overload: opportunities and challenges for the GP's desktop. AB - Recent developments in health data networks, the health sector and information systems, have created an overload of information available to the General Practitioner. The implementation of viable Health Data Networks within hospitals and subsequent connection to the GP's desktop PC enables increased access to patient records, decision-support and communication with experts around the world To address the high usage of expensive health services and lengthy waiting lists health services around the world are embracing programs such as Community, Coordinated, Shared and Managed care. This focus on coordination of care and increased emphasis on evidenced-based medicine is greatly enhanced through the advent of viable health data networks. Other resources such as databases, best practice guidelines, the web, email, telemedicine and a range of commercial programs that provide further services has created an overload of resources available to the GP. Current human-computer interface guidelines are not adequate for prescribing design solutions to deal with the information overload facing GPs. The challenge for the near future is to present the vast array of information sources to the GP in an acceptable and useable information system interface. Part of the solution may revolve around the development of standards for Electronic Health Record systems for GPs, as is being done currently in Australia; but we suspect that less mainstream interface technologies will be required to exploit the wealth of available healthcare information. PMID- 10384668 TI - Community medical care monitoring system for chronic patient. AB - The chronic disease usually can not be cured well but controlling by the daily care activity, regular diagnosis and dosage under the physician's instructions. In this research, we propose a community medical care monitoring model for monitoring and managing the chronic patients of community through the WWW (World Wide Web) service network and/or telephone voice service network. It can collect more patients' physiological data measured at home or community health agent. Therefore, the chronic disease patients will be getting better cure from his doctor's diagnosis due to the patient's effort. PMID- 10384669 TI - Development and evaluation of regional health database systems. AB - We have developed information systems for regional public health in some areas. These systems have the following functions: (1) to register results of examinations, life style, follow-up data, diseases, (2) to output some reports and statistics for daily jobs, (3) to make a personal health database, (4) to display stored data by character, trend graph, radar chart or bar chart, (5) to analyze regional health problems, (6) to extract dada about persons with some conditions, (7) to compare data among some examinations, (8) to analyze risk factors. Each database can be transferred to mini-notebook type personal computers. So, they can be used anywhere and any time. Some access controls are performed for security. The systems are used in routine works, health consultation for persons and health education for groups. We have evaluated the systems and got some useful results. Using the systems, daily jobs can be performed with good accuracy and less labor, each subject can understand one's health status easily and clearly, grasps of health problems can be made quantitatively and health projects can be planned by scientific methods. PMID- 10384670 TI - A hypermedia tool to support the interaction between radiologists and clinicians. AB - In this work we discuss problems related to the radiological reporting process and attempt to identify the ones which emerge during clinician-radiologist communication. We propose a hyper-reporting system, which allows the clinician to share the knowledge and critical factors that prompt the radiologist's diagnostic hypothesis during the discussion of results. This is possible by creating hypermedia links among different elements: reporting text, images, comments and the graphic models library. These elements make radiological knowledge explicit and clarify to the clinician the logical-diagnostic path traveled by the radiologist. A questionnaire, a survey of clinicians' and radiologists' needs allowed us to define some aspects of a hypermedia demo interface. Finally, we describe an example of a working session with the use of a few explanatory cards. PMID- 10384671 TI - Computer-aided documentation and therapy planning in pediatric oncology. AB - In the past 20 years considerable progress was made in pediatric oncology concerning the results of therapy in Germany. Nationwide multicentre trials provide protocols for the therapy of the children. The calculation of the chemotherapy according to the protocols is rather complex and the documentation efforts for the pediatricians are enormous. Thus, we developed as a first step an application system for Computer Aided Therapy Planning In Pediatric Oncology (CATIPO), that is in routine use in about 20 pediatric clinics in Germany. In order to support the physician comprehensively with the documentation that is necessary for the trials we currently develop a Documentation System for Pediatric Oncology (DOSPO) that comprises the functionality of chemotherapy planning according to the protocols released by the trial centres. Besides supporting the physician in clinical routine the major objective is to improve the quality of the documented data. To reach this aims DOSPO combines research aspects of decision support and clinical documentation: formal representation of general protocols, calculating of a particular therapy for a patient, data acquisition, communication interfaces for transferring the data to the trial centres. In order to support trial centres an authoring system and a central data dictionary will be developed. This will enable the trial centres to develop new modules for trial-specific data acquisition in the clinics. PMID- 10384672 TI - A generic model for Internet-accessed databases in epidemiology: a nutritional application. AB - The Web technology has made the exchange of information among heterogeneous platforms possible with no added cost to end-users. Adding to this, it made it possible to access databases residing on servers in different geographic locations giving thus conceptors and programmers the possibility of making distributed applications with a fraction of the time needed before. This technology is therefore very well suited to medical applications which are normally multi-user, multiplatform, and data-intensive. In this paper, we present an application for epidemiological studies based on the Web technology and the reuse of common concepts in epidemiology to produce a generic model that can be parameterisable by the end-user to suit the needs of his study. This model is then applied to develop an application to optimise the collection and processing of nutritional data for a nation-wide epidemiological study concerning renal failure patients. PMID- 10384673 TI - Nurses' requirements for information technology in the next millennium. AB - The main purpose of this study was to identify nurses' requirements for information technology in the next millennium. We distributed 350 questionnaires in a teaching hospital in Brazil and in the USA. We received 86 from Brazil and 78 from USA. Besides the capabilities of computers to be used in different sectors of human actions, nurses' requirements for information technology can not be considered sophisticated. Nurses in Brazil prefer a standalone system. In contrast, in USA they prefer systems that could be integrated to the whole healthcare system. Also, in the USA, nurses were more comfortable with use of computers than nurses in Brazil. In general terms, nurses in both countries feel that computer systems could make their practice easier and more efficient. PMID- 10384674 TI - Can the US minimum data set be used for predicting admissions to acute care facilities? AB - This paper is intended to give an overview of Knowledge Discovery in Large Datasets (KDD) and data mining applications in healthcare particularly as related to the Minimum Data Set, a resident assessment tool which is used in US long-term care facilities. The US Health Care Finance Administration, which mandates the use of this tool, has accumulated massive warehouses of MDS data. The pressure in healthcare to increase efficiency and effectiveness while improving patient outcomes requires that we find new ways to harness these vast resources. The intent of this preliminary study design paper is to discuss the development of an approach which utilizes the MDS, in conjunction with KDD and classification algorithms, in an attempt to predict admission from a long-term care facility to an acute care facility. The use of acute care services by long term care residents is a negative outcome, potentially avoidable, and expensive. The value of the MDS warehouse can be realized by the use of the stored data in ways that can improve patient outcomes and avoid the use of expensive acute care services. This study, when completed, will test whether the MDS warehouse can be used to describe patient outcomes and possibly be of predictive value. PMID- 10384675 TI - Intervention classifications for nursing practice and the ICNP: cultural considerations for Korean nurses. AB - There are several intervention classifications for nursing practice. Yet, their relevancy and clarity for international use has not been evaluated. Thus, the paper presents a review of 4 major nursing intervention classifications (Saba's Home Health Care Classifications, Omaha's Intervention Scheme, Grobe's Nursing Intervention Lexicon & Taxonomy, and the International Classification for Nursing Practice) for Korean nursing practice. A simple comparison procedure utilized to determine the relevancy and clarity of each overall scheme and specific elements. The findings showed that general concepts for the top level of all the classifications were both relevant and clear. However, specific elements of lower levels became less clear and relevant because of the lack of definitions and cultural differences. In particular, interventions related to psycho-social and environmental components appeared to be either unclear or non-relevant for Korean nursing practice while interventions related to physiological components were found to be very relevant and clear. In conclusion, multiple levels of hierarchical intervention schemes and the absence of definitions impede clarity of intervention classifications. In addition, for international use, an intervention classification needs to be sensitive to cultural relevancy in its elements. PMID- 10384676 TI - Experience journals: using computers to share personal stories about illness and medical intervention. AB - Medical advances make it increasingly possible for children with previously fatal illness to live and thrive. However, a significant number still experience repeated operations, hospitalization, and invasive procedures, or need special care at home. Many do so with little or no intervention to help them and their families cope with the emotional stresses involved. One significant source of emotional and cognitive support is the community of patients and families who have experienced similar medical procedures. However, in spite of a general willingness to share experiences, communication among patients and families is usually limited. To facilitate this process, we are investigating the use of computer technology to record, organize, and display stories about the experiences of families with children who have been treated for cardiac and neurological illness at Children's Hospital, Boston. We are asking children and their families to record text and multimedia vignettes describing some aspect of their illness, coping strategies, or care that might be useful to others. These contributions will be available for browsing at a secure World-Wide-Web site. However, economic realities preclude reliance on a professional site administrator to organize and monitor what we hope to be a rapidly growing Web site with a large, distributed authorship. The need to make the Web site fully accessible to users who have varying familiarity with computers and Web browsing imposes further constraints. We are therefore developing software to automate the process of managing and organizing an easily accessed Web site that contains an "Experience Journal." We describe this software, the rationale for its development, and our plans for its use in the coming year. PMID- 10384677 TI - A telecommunications system to manage patients with chronic disease. AB - The care of patients with chronic disease is a large and growing problem in the United States and other industrialized countries.' it is expensive, and the quality of care received by patients is often sub-optimal, resulting in poor health outcomes. We developed a totally automated computer-controlled telecommunications system, called TLC, that provides--frequent, close monitoring of patients with chronic disease and reports the results to the patients' physicians on a timely basis, so that they can intervene appropriately. TLC also monitors the patients' important self care activities, such as medication-taking, and provides education and counseling to improve the patients' performance of these activities. The system operates through regularly scheduled telephone conversations with patients' in their homes. An evaluation of a TLC chronic disease application for patients with hypertension demonstrated that use of the system was associated with significant improvement of the patients' adherence to their medication regimens and significantly improved blood pressure control. These results show that it is possible to design an information science-based health care delivery system that performs functions usually performed only by health care professionals, and suggests that information science will become an important means of delivering health care services in the next millennium. PMID- 10384678 TI - System approach for a multicenter, multilingual international telemedicine conference. AB - Chinese Telemed 96, a three way international telemedicine conference was successfully held in November 1996. Over 1,000 physicians, allied health professional and medical students participated in this conference in Beijing, Hong Kong and London. This program demonstrated that the quality of telecommunication technology was suitable for the Chinese University of Hong Kong, Faculty of Medicine to use it in facilitating medical consultations and sharing expertise among widely dispersed colleagues. The consensus of Hong Kong participants was that the audiovideo and telecommunication technology used in this conference were adequate. In order to improve overall quality, however, a better co-ordination of the technical support across sites is required. Other felt that the conference suffers from the shortage of highly trained technical staff required to support the communication system. However, following the total evaluation of this conference, it is concluded that while technical support, site and location are important factors in a good telemedicine conference, a well run telemedicine conference is more dependent on a sound planning process and its execution. PMID- 10384679 TI - Guidelines for screening for diabetic retinopathy revisited. PMID- 10384680 TI - Quality of life assessment in the collaborative ocular melanoma study: design and methods. COMS-QOLS Report No. 1. COMS Quality of Life Study Group. AB - The Collaborative Ocular Melanoma Study (COMS) is a set of randomized clinical trials sponsored by the National Eye Institute of the National Institutes of Health. The COMS is being conducted to evaluate the role of radiotherapy in the treatment of patients with choroidal melanoma. Primary choroidal melanoma can enlarge or metastasize and eventually cause death in a significant percentage of cases. The primary COMS trial is designed to determine whether enucleation (removal of the eye) or radiotherapy without removal of the eye provides the patient with the longest remaining lifespan. More than 40 clinical centers in the United States and Canada are participating in the COMS. The objective of the COMS is to assess the effect of treatment upon 5-year and 10-year survival and the reduction or elimination of the disease process in patients randomly assigned to receive either radiation or enucleation. An ancillary component of the COMS, referred to as the COMS-QOLS, was designed to measure the impact of disease and its treatment on quality of life. The two treatment approaches being investigated, enucleation and radiation therapy, are likely to have different psychological and physiological effects on the patients receiving them. The COMS QOLS assessments include the SF-36 Health Survey, the Activities of Daily Vision Scale (ADVS), the Visual Functioning Questionnaire (VFQ), and the Hospital Anxiety and Depression Scale (HADS). Patients are interviewed at selected intervals during follow-up; in addition, 200 patients will be interviewed before randomization and have repeat interviews at six months and annually after randomization. The patient's quality of life after treatment will become an important consideration in determining the best form of therapy, particularly in the event that no survival difference between treatment groups is found. PMID- 10384681 TI - Quality of life in patients with choroidal melanoma: a pilot study. AB - PURPOSE: To assess the internal consistency of a short, widely-used health related quality of life instrument in patients with choroidal melanoma, its appropriateness for use in a large clinical trial in patients with that disease, and the feasibility of various methods of instrument administration. METHODS: The SF-36 was administered to 31 choroidal melanoma patients identified from two ocular oncology practices. Fifteen patients were interviewed by telephone, 4 were interviewed in person at the clinic, and 12 patients completed the instrument themselves while at the clinic. RESULTS: Twenty-three patients had already been treated for choroidal melanoma; 20 patients were treated with some form of radiation. The remaining 8 patients had not yet been treated. Metastasis had not been diagnosed in any of the patients. Patient age ranged from 37 to 85 years with a median of 59 years; 55% of the patients were female. The in-person and telephone interviews performed better than self-administered interviews in terms of missing items due to non-response (0% vs. 3.2%). The overall SF-36 health profile for the choroidal melanoma patients was similar in shape to that expected for a general U.S. population sample with the same age and gender distribution, although the choroidal melanoma patients averaged 4 to 11 points higher, indicating better health-related quality of life, for all scales except the role functioning-emotional scale. CONCLUSIONS: The results supported the internal consistency of the SF-36 in this population. Interviewer administration of the SF 36 either in-person or by telephone was acceptable to the patients and provided more complete data. PMID- 10384682 TI - Prevention and control of epidemic keratoconjunctivitis in a teaching eye institute. AB - PURPOSE: To determine if an ongoing infection control program is associated with a reduction in rates of nosocomial outbreaks of epidemic keratoconjunctivitis (EKC) and outbreak morbidity from nosocomial EKC in a large teaching eye institute. METHODS: The number of nosocomial EKC outbreaks, the number of affected patients, and the total number of patient visits were collected for each year between 1984 and 1997. An infection control program was implemented in 1992. The program included specified methods of patient screening and isolation, handwashing, instrument disinfection, medication distribution, and furlough of infected employees. The program included two levels of intensity of infection control measures, for non-outbreak and outbreak conditions. We compared rates per 100,000 patient visits of nosocomial outbreaks of EKC and affected patients for the 6-year period after the program was implemented, 1992-1997, with corresponding rates for 1984-1991. RESULTS: One, to three nosocomial outbreaks of EKC occurred annually in the period 1984-1991. After the implementation of the infection control program, no nosocomial outbreaks occurred in three of six years studied. In the pre-infection control years 1984-1991, there were 3.89 outbreaks and 54.09 affected patients per 100,000 visits, respectively. For the post infection control years 1992-1997, the corresponding rates were 0.54 outbreaks and 5.66 affected patients per 100,000 patient visits. Rates for both outbreaks and affected patients were significantly lower for the post-implementation period (p < 0.005 and p < 0.0005, respectively). CONCLUSIONS: An ongoing infection control program was associated with decreased rates of nosocomial EKC outbreaks and outbreak morbidity from nosocomial EKC in our institute. Although several reports have described infection control measures that terminated individual outbreaks of nosocomial EKC, this study demonstrates that an ongoing infection control program may preemptively reduce nosocomial EKC outbreaks. PMID- 10384683 TI - Causes and prevalence of non-vision impairing ocular conditions among a rural adult population in sw Uganda. AB - Information is scanty about the extent of ocular morbidity in developing countries, particularly for non-vision impairing conditions (NVIC), although these constitute the majority of consultations in eye clinics. We have conducted a survey to determine the prevalence and causes of NVIC in a Ugandan adult population and compared our findings with the work pattern of the district hospital. Adults were screened using Snellen's illiterate E chart and those found with visual impairment (acuity less than 6/18) in either eye were referred to a low-vision clinic, and those with obvious ocular disease or symptoms, but without visual impairment, to an outreach clinic. A total of 2886 (53%) out of 5479 adults were screened. Of these, 257 (8.9%) were referred to the outreach clinic, of whom 173 (67%) attended. Of those attending the low-vision clinic 83% had visual impairment confirmed, and 92% of those attending the outreach clinic were confirmed not to have visual impairment. The four commonest NVIC observed at the outreach clinic were: presbyopia (48%), allergic conjunctivitis (20%), early cataract (9%) and infective conjunctivitis (8%), the same conditions as those most commonly seen at the district hospital. In this community, over 80% of NVIC are caused by four conditions which are potentially either correctable, operable or treatable. Training health workers to recognise and treat these conditions would serve the great majority of eye patients. Hospital activity data can be used cautiously to assess the relative frequency of NVIC in the community. PMID- 10384684 TI - Associations of performance-based and self-reported measures of visual function. The Beaver Dam Eye Study. AB - OBJECTIVE: To describe and compare performance-based and self-reported measures of visual function and to evaluate how each varied with age and how the performance-based measures were associated with the responses to self-reported questions about visual function. DESIGN: Population-based epidemiologic study. PARTICIPANTS: Adults participating in the first follow-up of the Beaver Dam Eye Study (n = 3,722). MAIN OUTCOME MEASURES: Performance-based measures of vision including current binocular, best-corrected and near visual acuities, sensitivity to light as measured by an automated perimeter, contrast sensitivity, and self reported measures from a standardized interview. RESULTS: All performance-based and self-reported visual functions decreased with increasing age (p < 0.0001 for each). On average, women did more poorly than men on all the performance-based measures (p < 0.0001 for each), and on some of the self-reported measures. For both genders, correlations among the performance-based measures ranged from 0.38 to 0.67. In general, individuals reporting good visual function or little difficulty with visual tasks scored better on performance-based measures compared to individuals whose questionnaire responses reflected limited visual function. Overall, about 39% of drivers reported that their vision caused them to limit their night driving. Correlations of performance-based with self-reported measures of visual function were greater for best-corrected, binocular and near visual acuity compared to correlations of visual sensitivity to light and contrast sensitivity. There were 1,285 persons whose best-corrected acuity was better than their current binocular acuity and there were 97 people with visual impairment considering only current binocular acuity whose vision improved to better than 20/40 after refraction. CONCLUSIONS: Performance-based and self reported measures of visual function were consistently and inversely related to age. Performance-based measures were, in general, more highly correlated between themselves than were self-reported measures. Self-reported visual functions were significantly positively correlated with all performance-based measures, but correlations were moderate. While high contrast measures are usually used in clinical settings and in many studies, the data suggest that in assessing function it is appropriate to measure several different aspects of vision, both performance-based and self-assessed. Our data give some indication of the impact these visual functions have on daily life as reflected by night driving and on near and distant visual acuity. New refractive corrections would likely improve the visual acuity in many older adults. PMID- 10384685 TI - Barriers to compliance with screening guidelines for diabetic retinopathy. AB - OBJECTIVE: To identify barriers to compliance with guidelines for diabetic retinopathy screening. METHODS: The population studied included 4410 adults, aged 31 to 64, enrolled in an Independent Practice Association (IPA) plan in Upstate New York, who were diagnosed with diabetes, and their Primary Care Physicians (408 PCPs). Claims data were used to calculate variables characterizing patients and their PCPs. Logistic regression models were estimated to identify factors associated with higher probability of screening. RESULTS: 34% of patients were screened in 1993. The probability of screening was significantly higher for older patients, for women, for patients who visit their PCPs more often and for those living in areas of higher average education and lower percentage of blacks. However, only 16% of diabetic patients received an annual screen in two consecutive years (1992 and 1993). The probability of consecutive annual screening was significantly associated only with gender and patient expenditures per month. CONCLUSION: The very low rate of diabetic retinopathy screening has implications for quality of life of patients with diabetes, long term costs of caring for them and social costs due to lost productivity. Interventions to increase screening rates are needed and should target both patients and their Primary Care Physicians. PMID- 10384686 TI - Prevalence of visual impairment in children: a review of available data. AB - Data on the prevalence, magnitude and causes of blindness and severe visual impairment in children are needed for planning and evaluating preventive and curative services for children, and for planning special education and low vision services. Prevalence data can be obtained from a variety of different sources, each of which has limitations. The available data suggest that there may be a ten fold difference in prevalence between the wealthiest countries of the world and the poorest, ranging from as low as 0.1/1000 children aged 0-15 years in the wealthiest countries to 1.1/1000 children in the poorest. In this paper, the available data are reviewed and the epidemiological methods and findings are discussed. PMID- 10384687 TI - The moving parts of voltage-gated ion channels. PMID- 10384688 TI - Trifluoroethanol and colleagues: cosolvents come of age. Recent studies with peptides and proteins. AB - Alcohol based cosolvents, such as trifluoroethanol (TFE) have been used for many decades to denature proteins and to stabilize structures in peptides. Nuclear magnetic resonance spectroscopy and site directed mutagenesis have recently made it possible to characterize the effects of TFE and of other alcohols on polypeptide structure and dynamics at high resolution. This review examines such studies, particularly of hen lysozyme and beta-lactoglobulin. It presents an overview of what has been learnt about conformational preferences of the polypeptide chain, the interactions that stabilize structures and the nature of the denatured states. The effect of TFE on transition states and on the pathways of protein folding and unfolding are also reviewed. Despite considerable progress there is as yet no single mechanism that accounts for all of the effects TFE and related cosolvents have on polypeptide conformation. However, a number of critical questions are beginning to be answered. Studies with alcohols such as TFE, and 'cosolvent engineering' in general, have become valuable tools for probing biomolecular structure, function and dynamics. PMID- 10384689 TI - [Etiology, pathogenesis and therapy of renal cell carcinoma]. AB - Renal cell carcinoma (RCC) of the kidney accounts fur 1-2% of all cancers and is the most common cancer arising in the adult kidney. The cause of RCC is not known. Several studies have shown a greater risk in smokers. Other factors are long-term dialysis and obesity, e.g. The morphological types of RCC have characteristic cytogenetic changes, but the details of genetic changes in renal tumorigenesis are not well understood. Approximately 13%-18% of patients with RCC have metastatic disease at initial presentation, and their prognosis remains unfavorable, because RCC is resistant to chemotherapy and radiotherapy. Only one third of patients with RCC cured, and strenous efforts are being made to optimize immunotherapy in patients with advanced or metastatic disease. PMID- 10384690 TI - [Pilot study on early diagnosis of renal cell carcinoma by sonography]. AB - The goal of this study was to investigate the practicability and effectiveness of a systematic screening for renal cell carcinoma by ultrasound in the course of the established German Health Ministry screening programs. METHODS: In two centers (Mainz and Wuppertal) a screening program for renal cell carcinoma for all citizens (age > 40 years) was established. The screening was divided into two phases (time period 1 year): (1) All citizens over 40 years could attend voluntarily a cost-free ultrasound investigation. (2) A follow-up investigation for the entire screening population was provided. RESULTS: Ten thousand volunteers attended the screening program. Mean age was 60 years. Thirteen renal cell carcinomas were detected. The incidence of other findings was 15%; none of those required further treatment. CONCLUSION: Systematic screening for renal cell carcinoma by ultrasound is cost-effective and showed high acceptance and practicability in a German population. The rate of detection of renal cell carcinoma was higher than initially statistically calculated. PMID- 10384691 TI - [Kidney spiral CT. Indication, method, results]. AB - The introduction on spiral computed tomography (spiral CT) has vastly enriched the methodological diversity of computer-tomographic scans. It allows for the recording of different perfusion or excretion stages of the kidney parenchyma of the urine draining paths by carrying out long-distance, phase-identical multiple examinations of the retroperitoneum. The description of the findings which are characterized by their local and contrasts behavior is possible. The following report describes the indications and technological process of kidney spiral CT using kidney-typical intravenous contrast media. Special emphasis is put on the advantages and limits of multiple phase spiral CT. Decisive preconditions are: 1. specific clinical query, 2. selection of the corresponding phase contrasts of the kidneys and uretra or bladder, 3. exact technical and temporal adjustment of the acquisition parameters. Scanning times are in the range of seconds. The overall examination can be carried out quick and without any major strain on the part of the patient. A sound proof and a general differentiation of focal kidney lesions can be derived from the acquired data. This is also true for kidneys and ureters findings. Bladder findings can be localized and differentiated according to stage. More than two "spiral acquisitions" should be carried out with re-straint taking exposure to radiation into account. Due to the sound registration of focal lesions, its capability of reproduction and its short-time examination, the spiral CT of the kidneys can be said to be the most effective current scanning method of the retroperitoneum following clinical examinations and sonography. PMID- 10384692 TI - [Angiography and interventional radiology of the kidney]. AB - For imaging of renal pathology a broad spectrum of radiologic diagnostic procedures are available which are, sometimes and particularly more recently, competing among each other in their diagnostic yield and relevance. For tumorous lesions ultrasound, computed tomography and magnetic resonance imaging are performed predominantly. Angiography is no longer required with the exception of highly selected cases and in some specific preoperative workup requirements. Until recently, catheter based digital subtraction angiography has been considered as gold standard. However, non-invasive techniques such as CT angiography and MR-angiography are evolving parallel to their quantum leap of resolutions and readiness to use. Nevertheless, well accepted criteria for quality assessement of these new modalities are still lacking. More comparison studies are urgently warranted. Despite the availability of ultrashort pulse sequences applying the T1 relaxation reduction effect of gadolinium enhanced MR techniques overestimation of renal artery stenosis still poses a substantial problem. Renal intervention implies a variety of procedures such as plain angioplasty, stent placement, embolization of traumatic and both benign and malignant tumors. These methods have emerged over the last two decades from a more experimental nature to a fully accepted treatment option. When renal artery angioplasty is embedded in an aggressive approach including stenting as an adjunct for more complex cases, renal ostial lesions and a well organized follow up regimen its therapeutic potential for treatment of renal insufficiency, malignant hypertension, for organ preservation bears a very high potential. Provided adequat periinterventional drug regimen restenosis rates may be as low as 10%. In highly selected cases capillary embolization might be used as an alternative to nephrectomy with a similar clinical outcome. Particularly the development of superselective small caliber embolization catheters parallel to further refinement of embolization material has aided to use superselective occlusion techniques in benign vascular lesions and renal trauma. PMID- 10384693 TI - [MRI of the kidneys. New diagnostic strategies]. AB - AIM: New diagnostic strategies for evaluation of the kidney by fast MR imaging techniques. MATERIAL AND METHODS: A comprehensive morphologic and functional evaluation of the kidney is proposed using fast MR imaging of renal morphology, multiphase 3D gadolinium MR angiography, MR urography and MR flow measurements. A single MR examination is designed to grade renovascular disease and assess the hemodynamic and functional significance, detect and characterize renal lesions and evaluate the urinary tract. RESULTS: The combined analysis of morphologic and functional data allows reliable assessment of renal artery stenosis, benign and malignant renal masses and diseases of the renal collecting system and ureters, as well as congenital abnormalities in good agreement to the results of conventional imaging modalities. The improved tissue contrast and additional functional information compensates for the disadvantage of a lower spatial resolution. CONCLUSION: Combined morphologic and functional MR examination represents a reliable, non-invasive and cost-effective alternative imaging modality for comprehensive diagnostic evaluation of renal disease. PMID- 10384694 TI - [Nuclear medicine in kidney diagnosis]. AB - A brief review of frequently requested examinations is presented. The clinical physiology associated with characteristic function images observed in obstruction, urinary leaks, hypertension, renal failure, and in renal transplantation are presented, as is the use of function images in the patient with renal malignancy. PMID- 10384695 TI - [Radiologic diagnosis of the kidneys in dialysis patients]. AB - The kidneys of patients with chronic renal failure undergoing maintenance hemodialysis may show different variances or complications. Most common are secondarily acquired renal cysts, which may be found in as many as 92% of patients after 8 years of hemodialysis. Single (in 12.5% of patients) or multiple (8.3%) cysts with bleeding are common; additionally, hematuria or ruptured cysts may be found. Bleeding into cysts is more common in patients with autosomal dominant polycystic kidney disease. Due to the decreasing urinary production development of kidney stones is very uncommon, but calcifications in or around cysts can be found in 71% of patients. Kidney tumors occur 41 times more often in patients with chronic renal failure than in patients without kidney disease. We detected tumors in 4.2% of our patients on long-term dialysis. Diagnostic differentiation of the relatively slow growing and fairly late metastasizing malignant tumors from adenomas is not possible. Nevertheless, we screen our patients every 3-4 years. Computed tomography is superior to ultrasonography for this purpose, because ultrasonography lacks the necessary sensitivity in this group of patients. PMID- 10384696 TI - [Radiodiagnosis following kidney transplantation]. AB - Diagnostic imaging after renal transplantation is of high importance in the differential diagnosis of peri- and postoperative complications. Sonography with color duplex imaging is the method of choice in the diagnosis of acute transplant rejection. MRI is an additional method in the diagnosis of transplant dysfunction especially in diagnosis of perirenal fluid collections. MR angiography and MR urography are noninvasive diagnostic modalities with the potential to replace angiography and pyelography. Angiography, complemented by carbon dioxide angiography, still is the gold standard in the diagnosis of transplant artery stenosis. PMID- 10384697 TI - [Diagnostic imaging of the kidneys and the urinary tract in children]. AB - Imaging flow charts differ in pediatric and general radiology. The reasons are: different illnesses, different consequences arising out of imaging results and different sequence of imaging methods. Ultrasound is always the first imaging method of the urinary tract in infancy and childhood starts with ultrasound with the exception of severe abdominal trauma which is investigated by computer tomography. The decision "normal or abnormal" is possible using ultrasound in the most pediatric cases. The diagnostic value and significance of ultrasound in infancy and childhood is far better than in general radiology because of the higher resolution of the high-frequency units taken. The result of the ultrasound examination should be the basis for the following imaging procedures. We will describe diagnostic flow charts starting with three important clinical symptoms: Prenatal pathology, urinary tract obstruction and urinary tract infection. PMID- 10384698 TI - [Mn-TPPS4 in the diagnosis of malignant skin tumors. In vivo studies with high resolution magnetic resonance tomography in melanotic melanoma]. AB - PURPOSE: Some authors postulate an avidity of certain porphyrin derivatives for tumors. The aim of this study was to examine the contrast enhancement of implanted melanotic melanoma after application of Mn-TPPS4 to achieve a better characterization of this malignant skin tumor. MATERIAL AND METHOD: High resolution MR imaging (2.0 Tesla, 2.0-cm surface-coil, T1-weighted FLASH-2D sequence) was performed on 15 mice (C57b16) with intracutaneous implanted melanoma (B16F1), before and after intravenous administration of either Gd-DTPA (Magnevist, Schering, Berlin) as a reference contrast medium, or Mn-TPPS4 (Porphyrin Products, Schering, Berlin). The images were evaluated quantitatively by calculating percentage enhancement, slope of signal intensity-to-time curves, percentage increase of the signal intensity, signal-to-noise and contrast-to noise-ratios. The qualitative evaluation was accomplished by visual assessment of the enhancement, the demarcation of the tumors from the surrounding tissue and the homogeneity of the tumors. RESULTS: Contrast medium-enhanced images showed an increased signal intensity from all tumors with no signifikant difference between the contrast media. Demarcation of tumors from the surrounding tissue was better following administration of contrast media; regressive and altered areas was more clearly depicted. CONCLUSION: A specific accumulation of the metalloporphyrin Mn TPPS4 in melanotic melanoma could not be proved. Based on the small and insignifikant differences in the results obtained with the two contrast media, and on the side effects of the metalloporphyrin, the usefulness of Mn-TPPS4 as a contrast medium for MRT is limited. PMID- 10384699 TI - [Juvenile shoulder injury]. PMID- 10384700 TI - [Radiology of the manual skeleton. 1. Inflammatory joint diseases and rheumatology]. PMID- 10384701 TI - [Therapeutics available in veterinary medicine for animals used in food production]. PMID- 10384702 TI - [Case report. Colic symptoms in a cow]. PMID- 10384703 TI - [Diagnosis and epidemiology of Neospora caninum-associated abortions in cattle]. AB - Neospora caninum, a recently discovered protozoan parasite closely related to Toxoplasma gondii, has world-wide been recognized as an important cause of bovine abortion. N. caninum possesses a wide host range. The dog can be a definitive host for N. caninum. In cattle, the infection is transmitted transplacentally with high efficiency, while the majority of congenitally infected calves is clinically normal at birth and thereafter. Whether horizontal transmission occurs in cattle and whether this potential mode of transmission has epidemiological significance, remains to be elucidated. N. caninum-associated abortions can occur in epidemic or endemic form in a herd. The clinical symptoms of bovine neosporosis are confined to the occurrence of abortion, stillbirth and weak calves. Multifocal nonsuppurative encephalitis represents the most frequent pathohistological finding in N. caninum-associated abortions. The causative agent can be demonstrated by immunohistochemistry or polymerase chain reaction. Serological techniques can be used for indirect diagnosis. On the basis of the available diagnostic methods and the present knowledge about the epidemiology of the infection proposals are made regarding diagnosis, epidemiological assessment and prophylaxis of N. caninum-associated abortion problems in cattle herds. PMID- 10384704 TI - [Reaction of the pituitary gland to a single GnRH administration in the dependence of energy balance in cattle in the puerperium]. AB - After reviewing the relevant literature about the relationship of negative energy balance post partum and the onset of cyclicity the aim of this study will be described. The objective of this trial was to evaluate the influence of negative energy balance on the sensitivity of the pituitary for GnRH. Two groups of Holstein Friesian cows were randomly treated either with GnRH (Fertagyl) or a placebo. Considering the weekly calculated negative energy balance, the reaction of the pituitary was determined by analyzing LH in 16 blood samples, collected every 20 minutes starting one hour before and ending four hours after treatment. All cows treated with GnRH showed an intensive LH-surge, significantly different from the controls. These data suggest, that the sensitivity of the pituitary gland is not directly influenced by negative energy balance. The hypothalamus has the inhibiting priority over the pituitary delaying post partum cyclicity. PMID- 10384705 TI - [Investigations on the behavior of C-reactive protein, cell count, lactose content as well as electric conductivity in quarter milk samples of subclinically diseased quarters of the udders in relation to bacteriologic results]. AB - Inflammations of the udder influence the amount of somatic cells and lactose as well as electrical conductivity in milk. They are often caused by bacterial agents. Similar reactions are to be seen in relation to C-reactive protein (CRP). The amount of CRP in milk depends on concentration and characteristics of the agents and the phase of inflammation. We found a significant increase of CRP in milk samples with Streptococcus (Sc.) uberis. There was no significant difference in CRP concentrations in samples with coagulase positive Staphylococcus spp. (CPS) and bacteriological negative samples and such with Corynebacterium (C.) bovis, Micrococcus spp. and Staphylococcus (S.) epidermidis. We only found high correlations to the amount of somatic sells and lactose as well as electrical conductivity in samples with Sc. uberis. The results of our investigations show that CRP may be a parameter for better understanding subclinical inflammations of udder. PMID- 10384706 TI - [Comparative investigations of a combined vaccine against parvovirus and erysipelas and corresponding monovaccines in different vaccination schedules. 1: Field trial]. AB - In a field trial, the development of antibodies of a combined vaccine against the porcine parvovirus (PPV) as well as against swine erysipelas was compared with corresponding mono vaccines. Furthermore, these vaccines were used in different vaccination schedules. The tests were carried out on 109 gilts in three closed farms. In all gilts, a basic immunization repeated twice was carried out at the age of six months and at intervals of three weeks. The revaccination was carried out four months after the basic immunization with half of the animals, and six months after the basic immunization with the remaining gilts. Between the combined vaccine and the mono vaccine no significant differences in the development of antibodies against PPV could be found according to different vaccination schedules. The gilts having been vaccinated with the mono vaccine and boostered six months later showed significantly higher antibody titers against Erysipelothrix rhusiopathiae. Between the remaining vaccination groups no significant difference in the development of the antibodies against swine erysipelas could be found. On only one farm, a continuous decrease of antibody titers against PPV in case of altogether 238 non-vaccinated piglets until the sixth month of life could be observed. On the two other farms, an increase of antibody titers against PPV could be found at different points of time, which indicates an infection of the piglets. Between the individual vaccination groups no significant antibody titers against PPV could be measured in milk tests. With regard to the number of piglets born alive per litter, the number of piglets born dead per litter and the number of mummies, a significant difference could neither be found between the vaccination groups 1-4. PMID- 10384707 TI - [Salivary cortisol as a stress parameter in piglets]. AB - The relationship between salivary and plasma levels of total and free cortisol was monitored in 97 male piglets, aged two to four weeks, subjected to castration. Samples were taken 10 minutes before (basal value) as well as one, two, three, four and 24 hours post castration and at the same time intervals from a control group of 17 animals which did not undergo surgery. Simultaneously to blood (indwelling catheter) withdrawing saliva was collected by two cotton swabs. Cortisol levels were measured by radioimmunoassay (RIA). A highly significant increase in total, free and salivary cortisol was found within the first four hours after castration compared to the control group. The percentage increase one hour after castration above basal values was highest in free plasma cortisol (21.08 +/- 2.03 nmol/l vs. 61.26 +/- 4.16 nmol/l; 290.6%), and lowest in total plasma cortisol (177.33 +/- 9.69 nmol/l vs. 374.09 +/- 18.21 nmol/l; 211.0%), whereas salivary cortisol showed an 255.7% increase (10.46 +/- 1.03 nmol/l vs. 26.75 +/- 1.93 nmol/l). Total cortisol included 11.9-16.4% free cortisol. Salivary cortisol concentration was between 5.9% and 7.5% of the total plasma cortisol concentration. The highest correlation between total plasma cortisol and salivary cortisol occurred one hour after castration (r = 0.57; p < 0.01). The correlation between free and salivary cortisol was lowest for basal values (r = 0.27; p < 0.05), whereas correlations for the remaining time points were highly significant (0.41 < or = r < or = 0.61; p < 0.01). For the control group significant correlations were found between salivary and total plasma cortisol (0.58 < or = r < or = 0.89; p < 0.05) and between free and salivary cortisol (0.63 < or = r < or = 0.92; p < 0.05). The present work indicates that the measurement of salivary levels of cortisol reflects the concentration of this hormone in plasma samples of piglets. PMID- 10384708 TI - [Diagnosis, therapy and endocrinologic parameters of persistent follicles in mares in comparison with preovulatory follicles]. AB - During the 1997 breeding season persistent follicles were diagnosed in 17 mares. In 16 of these mares a total of 17 follicles were transabdominally punctured and the steroids oestradiol, progesterone and testosterone were measured in the follicular fluid and in blood serum. In ten mares serving as a control group preovulatory follicles were punctured. The follicular fluid of the persistent follicles revealed a very high variability of the steroid concentrations. Depending on the steroid ratio within the follicles, eight follicles were rated as being intact, three follicles were undergoing atresia and five follicles were luteinized. Because of the high oestradiol levels of the follicular fluid within the control group, all of these follicles were considered to be intact. In both groups, no correlation of the steroid concentration between serum and follicular fluid was detectable. This fact argues against a passive diffusion of the steroids through the follicular wall. By puncturing the persistent follicles it was possible to bring the affected mares back into a physiological oestrus cycle within a normal dioestrus period. PMID- 10384709 TI - [Liver fluke infestation in New World camelids. Parasitology, pathology, clinical findings and therapy]. AB - In Llamas and Alpacas infestation with Fasciola hepatica or Dicrocoelium dendriticum can cause liver damage, sometimes even with lethal outcome. Once infected South American Camelids (SACs) react more sensitively to these parasites than other domestic ruminants. We report here on the pathology, parasitology, clinics and therapy of this disease. Concerning Dicrocoelium dendriticum we describe own clinical results and therapeutic outcome in addition to the pathological investigation. According to anatomic corrosion casts, the bile ducts of SACs show more similarity with the equine bile system than with the bile system of domestic ruminants. PMID- 10384710 TI - The development of resistance to caffeine in Drosophila prosaltans: productivity and longevity after ten generations of treatment. AB - The productivity of Drosophila prosaltans treated with six concentrations of caffeine (from 50 micrograms/ml to 2,500 micrograms/ml of culture medium) during ten generations (approximately 8 months) decreased in a dosage dependent manner in every generation, but at the end of the treatment the flies in all experiments recovered normal productivity, except for those treated with 2,500 micrograms/ml. Longevity in the tenth generation was significantly reduced in males and females only in the 2,500 micrograms/ml dosage, with males being much more affected than females. In a previous study in which the treatment was done in a single generation, productivity exhibited only a partial recovery when the treatment ceased and longevity was significantly reduced in 1,500 micrograms/ml dosages. The hypothesis of selection occurring in ten generations leading to recovery in productivity and to a reduction in the processes which cause a decrease in longevity is being considered. PMID- 10384711 TI - Esterase patterns and phylogenetic relationships of species and strains included in the Drosophila buzzatii cluster. AB - Ten strains of two species in the Drosophila buzzatii cluster (D. serido and D. seriema) were examined as to esterase patterns using polyacrylamide gel electrophoresis. The migration rate of esterases, and their substrate specificity to alpha and beta naphthyl acetates, were analysed. Other esterase features such as inhibition behaviour, presence in males and females and location in the head, thorax or abdomen of flies, were also examined. The present data, together with results obtained by others for eight strains of D. koepferae, D. serido, D. seriema and D. buzzatii, show that 69 bands have been detected in the eighteen strains studied. This total number of bands was used for comparison of strains and species by similarity index, analysis of dependence and cluster analysis. The comparisons confirmed the existence of a high degree of similarity among D. seriema strains and among D. koepferae strains, but indicated differentiation among the D. serido strains. Two strains (D69R2 and D69R5) which differed from the others of the latter species, showed closer affinities with D. buzzatii, which indicates the need for further work on those strains classified as D. serido. PMID- 10384712 TI - Synthesis of 4-alkyl (aryl)-6-aryl-3-cyano-2(1H)-pyridinones and their 2-imino isosteres as nonsteroidal cardiotonic agents. AB - Thirty new 1,2-dihydropyridine derivatives of the general formula 4-alkyl (aryl) 6-aryl-3-cyano-2(1H)-pyridinones (1-15) and 4-alkyl (aryl)-6-aryl-3-cyano-2(1H) iminopyridines (16-30) were synthesized using one-pot multicomponent reactions of the properly substituted acetophenone, appropriate aldehyde, ammonium acetate and ethyl cyanoacetate (1-15) or malononitrile (16-30) in ethanol. These target compounds (1-30) were evaluated for their cardiotonic activity using the spontaneously beating atria model, from reserpine-treated guinea pigs. The best pharmacological profile was obtained with 3-cyano-6-(3,4-dimethoxyphenyl)-4-(4 hydroxyphenyl)-2(1H)-pyridinone (9) which displaced selectivity for increasing the force of contraction (108.7 +/- 6.7,% change over control) rather than the frequency rate (40.8 +/- 5.3,% change over control) at a 5 x 10(-4) M concentration. The effects of structural changes upon activity are reported. PMID- 10384713 TI - Synthesis and structure-activity relationship studies of new endothelin pseudopeptide analogues containing alkyl spacers. AB - We replaced the Asp18-Ile19 dipeptide of the C-terminal ET analogue Ph-Ph-CH2-O N=CH-CO-Phe-Asp-Ile-Ile-Trp-OH by alkyl spacers of various lengths to investigate the role of the aminoacidic central portion of the molecule and to define the N terminal and C-terminal pharmacophoric regions of this analogue. The side-chains of the central dipeptide have been shown to be irrelevant for the binding of the molecule to the receptor, but the distance between the two postulated sites of interaction of the ligand with the ETB receptor appears to be fundamental. PMID- 10384714 TI - Synthesis and analgesic activity of some triazoles and triazolothiadiazines. AB - The synthesis of some triazoles and triazolothiadiazines starting from (5,6,7,8 tetrahydronaphthalen-2-yl)oxyacetic acid is described. The chemical structure of the compounds were elucidated by analytical, IR, 1H NMR and mass spectral studies. Some of the newly synthesized compounds were tested for analgesic activity and compounds 5b, 5c, and 5d exhibited promising analgesic activity. PMID- 10384715 TI - Synthesis and antimicrobial properties of cephalosporin derivatives substituted on the C(7) nitrogen with arylmethyloxyimino or arylmethyloxyamino alkanoyl groups. AB - Some 7-aminocephalosporanic acid (7-ACA) derivatives substituted on the C(7) nitrogen with 2-(arylmethyloxyimino)propionyl (3a-f), 2 (arylmethyloxyamino)propionyl (4a-d) and (arylmethyloxyamino)acetyl (2a-d) moieties were synthesized by reaction of the appropriate acylating agents with 7 ACA protected as a t-butyl ester, followed by removal of the t-butyl protecting group. The new compounds, tested in vitro for their antimicrobial activity against Gram-positive and Gram-negative bacteria, proved to possess a modest activity directed only against Gram-positive microorganisms. PMID- 10384716 TI - Gold(I) complexes as antimicrobial agents. AB - Seven gold complexes were prepared and investigated for biocidal activity against Gram-positive and -negative bacteria, fungi and protozoa. All of them were active against the tested microorganisms with the exception of Pseudomonas aeruginosa. In many, cases minimum inhibitory concentrations (MIC) were well below 1 microgram/ml. The activity is not simply related to the gold content, but also to the nature of both the phosphine and the aminothiol to which the metal is bound. PMID- 10384717 TI - Triorganotin compounds as antimicrobial agents. AB - Six triorganotin derivatives of thiolupinine(1-mercaptolupinane), 2 mercaptobenzoxazole and 2-mercaptobenzothiazole were prepared and tested against several bacteria, fungi and protozoa. Most compounds exhibited high activity against the tested microorganisms and particularly worth noting was the activity of triethyltin lupinylsulfide on Gram-negative strains. Triethylgermanium lupinylsulfide was also prepared but was devoid of action on the whole set of tested microorganisms. PMID- 10384718 TI - Synthesis and HIV-1 inhibitory properties of new tetrahydrobenzoquinazolinedione and tetrahydrobenzocycloheptenuracil derivatives and of their thioxo analogues. AB - Some new tetrahydrobenzoquinazolinediones 2a-4a, tetrahydrobenzocycloheptenuracils 5a, 6a and their thioxo analogues 2b-6b were synthesized within a project aimed at obtaining new HIV-1 tricyclic inhibitors whose scaffold includes a pyrimidine and a phenyl ring, which are present in various HIV-1 non-nucleoside inhibitors. Among the tetrahydrobenzoquinazolinediones 2a-4a, compounds 3a and 4a, in which the tricyclic system is respectively in an angular or linear arrangement, proved to possess a HIV-1 inhibitory activity which was in the micromolar range, while compound 2a, in which the tricyclic system is in the angular arrangement opposite to that of 3a, was found to be completely inactive. As regards the tetrahydrobenzocycloheptenuracil derivatives (5a and 6a), only 5a showed an inhibitory activity similar to that of 3a and 4a. Furthermore, all thioxo analogues 2b-6b were found to be devoid of any activity. PMID- 10384719 TI - Quinolizidinyl derivatives of 2,3-dihydro-2-oxo-1H-benzimidazole-1-carboxylic acid and 1-homolupinanoyl benzimidazolones as ligands for 5-HT3 and 5-HT4 receptors. AB - Five 2,3-dihydro-2-oxo-1H-benzimidazole-1-carboxylic acid derivatives of lupinine, epi-lupinine and lupinylamine, together with two 1-homolupinanoyl benzimidazolones were prepared and investigated for their ability to displace specific radioligands from 5-HT3 and 5-HT4 receptors. The synthesized compounds were only moderately active, with IC50 in the micromolar range. The compound with the highest affinity for 5-HT4 receptor was tested for the enhancement of intestinal transit rate but was inactive at the oral dose of 100 mg/kg. PMID- 10384720 TI - Synthesis and anti-HIV-1 activities of new pyrimido[5,4-b]indoles. AB - A set of new pyrimido[5,4-b]indole derivatives that are structurally related to some non-nucleside HIV-1 reverse transcriptase inhibitors were synthesized and biologically evaluated for their activity as inhibitors of wild and mutant HIV-1 RT types in an 'in vitro' recombinant HIV-1 RT screening assay, as well as anti infectives in HLT4lacZ-1IIIB cells. Preliminary structure-activity relationships suggest that activity is promoted by simultaneous substitution in positions 2 and 4, especially when chains of alkyldiamine type are present, and by electron releasing substituents (methoxy) in positions 7 and 8. The inactivity or the very low activity of title derivatives does not suggest interest in AIDS therapy. PMID- 10384721 TI - Factors influencing imitation of manipulatory actions in chimpanzees (Pan troglodytes). AB - The purpose of the study was to investigate what kind of factors determine the degree of difficulty for chimpanzees (Pan troglodytes) when they imitate actions. Five adult chimpanzees were instructed to perform 48 arbitrary manipulatory actions consisting of different bodily motor patterns and object directionality. Results showed that actions in which an object is directed toward another external location (another object and one's own body) were easier to perform than those that involved manipulating a single object alone. Actions involving unfamiliar motor patterns were more difficult to perform than those involving familiar motor patterns that were already present in the subject's repertoire. Error responses were characterized as perseverative repetition of previously instructed actions. These findings suggest that chimpanzees find the directionality of manipulated objects a more salient cue than details of the demonstrator's body movements performing the manipulation. PMID- 10384722 TI - Strategies used to combine seriated cups by chimpanzees (Pan troglodytes), bonobos (Pan paniscus), and capuchins (Cebus apella). AB - The authors investigated strategies used to combine seriated cups by apes (Pan troglodytes and P. paniscus) and monkeys (Cebus apella) using a protocol reported in P. M. Greenfield, K. Nelson, and E. Saltzman's (1972) study with children. It was hypothesized that apes would exhibit more hierarchical combinations of cups than monkeys, given apes' language capacity, and that apes would seriate the cups more efficiently than monkeys. As predicted, apes made many structures with the cups using a variety of strategies, and monkeys rarely combined the cups. After a training phase to orient monkeys to the task, the 2 genera did not differ in the strategies used to combine the cups or in efficiency in seriating the cups. Success in this task suggests that sensorimotor versions of hierarchically organized combinatorial activity are well within apes' and monkeys' abilities. PMID- 10384723 TI - Structural characterization of triacylglycerols as lithiated adducts by electrospray ionization mass spectrometry using low-energy collisionally activated dissociation on a triple stage quadrupole instrument. AB - We describe features of tandem mass spectra of lithiated adducts of triacylglycerol (TAG) species obtained by electrospray ionization mass spectrometry (ms) with low-energy collisionally activated dissociation (CAD) on a triple stage quadrupole instrument. The spectra distinguish isomeric triacylglycerol species and permit assignment of the mass of each fatty acid substituent and positions on the glycerol backbone to which substituents are esterified. Source CAD-MS2 experiments permit assignment of double bond locations in polyunsaturated fatty acid substituents. The ESI/MS/MS spectra contain [M + Li - (RnCO2H)]+, [M + Li - (RnCO2Li)]+, and RnCO+ ions, among others, that permit assignment of the masses of fatty acid substituents. Relative abundances of these ions reflect positions on the glycerol backbone to which substituents are esterified. The tandem spectra also contain ions reflecting combined elimination of two adjacent fatty acid residues, one of which is eliminated as a free fatty acid and the other as an alpha, beta-unsaturated fatty acid. Such combined losses always involve the sn-2 substituent, and this feature provides a robust means to identify that substituent. Fragment ions reflecting combined losses of both sn-1 and sn-3 substituents without loss of the sn-2 substituent are not observed. Schemes are proposed to rationalize formation of major fragment ions in tandem mass spectra of lithiated TAG that are supported by studies with deuterium labeled TAG and by source CAD-MS2 experiments. These schemes involve initial elimination of a free fatty acid in concert with a hydrogen atom abstracted from the alpha-methylene group of an adjacent fatty acid, followed by formation of a cyclic intermediate that decomposes to yield other characteristic fragment ions. Determination of double bond location in polyunsaturated fatty acid substituents of TAG is achieved by source CAD experiments in which dilithiated adducts of fatty acid substituents are produced in the ion source and subjected to CAD in the collision cell. Product ions are analyzed in the final quadrupole to yield information on double bond location. PMID- 10384724 TI - Distinction among isomeric unsaturated fatty acids as lithiated adducts by electrospray ionization mass spectrometry using low energy collisionally activated dissociation on a triple stage quadrupole instrument. AB - Features of tandem mass spectra of dilithiated adduct ions of unsaturated fatty acids obtained by electrospray ionization mass spectrometry with low-energy collisionally activated dissociation (CAD) on a triple stage quadrupole instrument are described. These spectra distinguish among isomeric unsaturated fatty acids and permit assignment of double-bond location. Informative fragment ions reflect cleavage of bonds remote from the charge site on the dilithiated carboxylate moiety. The spectra contain radical cations reflecting cleavage of bonds between the first and second and between the second and third carbon atoms in the fatty acid chain. These ions are followed by a closed-shell ion series with members separated by 14 m/z units that reflect cleavage of bonds between the third and fourth and then between subsequent adjacent pairs of carbon atoms. This ion series terminates at the member reflecting cleavage of the carbon-carbon single bond vinylic to the first carbon-carbon double bond. Ions reflecting cleavages of bonds distal to the double bond are rarely observed for monounsaturated fatty acids and are not abundant when they occur. For polyunsaturated fatty acids that contain double bonds separated by a single methylene group, ions reflecting cleavage of carbon-carbon single bonds between double bonds are abundant, but ions reflecting cleavages distal to the final double bond are not. Cleavages between double bonds observed in these spectra can be rationalized by a scheme involving a six-membered transition state and subsequent rearrangement of a bis-allylic hydrogen atom to yield a terminally unsaturated charge-carrying fragment and elimination of a neutral alkene. The location of the beta-hydroxy-alkene moiety in ricinoleic acid can be demonstrated by similar methods. These observations offer the opportunity for laboratories that have tandem quadrupole instruments but do not have instruments with high energy CAD capabilities to assign double bond location in unsaturated free fatty acids by mass spectrometric methods without derivatization. PMID- 10384725 TI - Tandem mass spectrometry of model peptides modified with trans-2-hexenal, a product of lipid peroxidation. AB - Small molecules formed during lipid peroxidation can react with the basic groups in proteins through different mechanisms. Recently, substituted pyridinium moieties were observed during in vitro incubations of lysine-containing peptides with 2-alkenals. To explore the dissociation behavior of peptides with pyridinium derivatized lysine residues, the peptide ions created through either matrix assisted laser desorption/ionization or electrospray ionization were studied with tandem mass spectrometry. The permanently charged pyridinium ions fragment primarily through the charge-remote processes. Under high energy collision induced dissociation, a number of diagnostic ions were observed that could potentially be used to identify modified residues in proteins. The origins of these ions were studied using deuterium exchange and higher-order mass spectrometry experiments using an ion trap instrument. Rational structures for these ions are proposed. PMID- 10384726 TI - Effects of salt concentration on analyte response using electrospray ionization mass spectrometry. AB - The effect of salt concentration on analyte response using electrospray ionization mass spectrometry (ESI-MS) was measured and compared to that predicted by Enke's equilibrium partitioning model. The model predicts that analyte response will be proportional to concentration and that the response factor will decrease with increasing electrolyte concentration. The measured analyte response is proportional to concentration over four orders of magnitude when the electrolyte concentration is below 10(-3) M, as the model predicts. The concentration of excess charge ([Q]) generated by the ESI process increases significantly at 10(-3) M ionic concentration, but the response factor decreases at this concentration. Changes in shape of the spray that cause a loss of ion transmission efficiency may be the basis for the decrease in response. An increase in the analyte response factor with increasing electrolyte concentration is observed for electrolyte concentrations below 10(-3) M. An explanation for this based on the electrical double layer is proposed. PMID- 10384727 TI - Binding of aldose reductase inhibitors: correlation of crystallographic and mass spectrometric studies. AB - Aldose reductase is a NADP(H)-dependent enzyme, believed to be strongly implicated in the development of degenerative complications of Diabetes Mellitus. The search for specific inhibitors of this enzyme has thus become a major pharmaceutic challenge. In this study, we applied both X-ray crystallography and mass spectrometry to characterize the interactions between aldose reductase and four representative inhibitors: AminoSNM, Imirestat, LCB3071, and IDD384. If crystallography remains obviously the only way to get an extensive description of the contacts between an inhibitor and the enzymatic site, the duration of the crystallographic analysis makes this technique incompatible with high throughput screenings of inhibitors. On the other hand, dissociation experiments monitored by mass spectrometry permitted us to evaluate rapidly the relative gas-phase stabilities of the aldose reductase-inhibitor noncovalent complexes. In our experiments, dissociation in the gas-phase was provoked by increasing the accelerating voltage of the ions (Vc) in the source-analyzer interface region: the Vc value needed to dissociate 50% of the noncovalent complex initially present (Vc50) was taken as a gas-phase stability parameter of the enzyme inhibitor complex. Interestingly, the Vc50 were found to correlate with the energy of the electrostatic and H-bond interactions involved in the contact aldose reductase/inhibitor (Eel-H), computed from the crystallographic model. This finding may be specially interesting in a context of drug development. Actually, during a drug design optimization phase, the binding of the drug to the target enzyme is often optimized by modifying its interatomic electrostatic and H bond contacts; because they usually depend on a single atom change on the drug, and are easier to introduce than the hydrophobic interactions. Therefore, the Vc50 may help to monitor the chemical modifications introduced in new inhibitors. X-ray crystallography is clearly needed to get the details of the contacts and to rationalize the design. Nevertheless, once the cycle of chemical modification is engaged, mass spectrometry can be used to select a priori the drug candidates which are worthy of further crystallographic investigation. We thus propose to use the two techniques in a complementary way, to improve the screening of large collections of inhibitors. PMID- 10384728 TI - Characterization of tt15, a novel transparent testa mutant of Arabidopsis thaliana (L.) Heynh. AB - The Arabidopsis thaliana seed coat typically has a brown color due to the accumulation of flavonoid pigments in the testa. Mutants of A. thaliana with defects in pigment biosynthesis often produce seeds that are olive brown or even yellow in appearance, and the responsible genetic loci are referred to as TRANSPARENT TESTA (TT). Large-scale screening for mutants affected in seed development and complementation analysis of a candidate mutant line with all published A. thaliana tt mutants identified a new tt locus designated tt15. The tt15 mutation maps to the lower part of chromosome 1. Mutant plants produced pale greenish-brown seeds whose dormancy was slightly reduced. The phenotype was consistent with the maternal origin of the testa. Analysis of pigment accumulation and the study of expression patterns of genes involved in flavonoid biosynthesis in tt15 plants and seeds indicated a seed-specific phenotype. Most notable was a reduction of the cyanidin and quercetin content of tt15 seeds. PMID- 10384730 TI - Synthesis and oxidative insolubilization of cell-wall proteins during osmotic stress. AB - The cell walls in the new white roots of jack pine (Pinus banksiana Lamb.) were observed to constrict around the shrinking protoplast of osmotically stressed roots, and pressure was maintained via an apparent adjustment of cell-wall size and elasticity. These elastic alterations of the cell wall permitted the root cells to maintain full turgor despite the loss of most of the water in the tissue. The constriction of the root cell wall around the dehydrating protoplasts to maintain turgor may reflect changes in cell wall structure. We found that these shrinking root cells synthesize and secrete into the intercellular fluid a set of proteins. These proteins become tightly associated (i.e. guanidine HCl- and sodium dodecyl sulfate-insoluble) with the cell wall but can be released from the matrix, after briefly boiling in 0.1% sodium dodecyl sulfate, by the combination of guanidine HCl, CaCl2 and dithiothreitol. However, these cell-wall proteins became insoluble with time. The proteins could subsequently be destructively extracted from the wall with acid NaClO2 treatments. After these proteins were incorporated into the cell walls, the roots adopted a new, smaller maximal tissue volume and elastic coefficients returned to normal levels. PMID- 10384732 TI - SbRLK1, a receptor-like protein kinase of Sorghum bicolor (L.) Moench that is expressed in mesophyll cells. AB - To study the metabolic interactions between mesophyll and bundle-sheath cells of C4 plants, protein kinases possibly involved in the regulatory processes and signal transduction pathways have been cloned and characterized. A receptor-like protein kinase (RLK) cDNA clone from the C4 plant Sorghum bicolor (L.) Moench has been identified. The deduced protein was designated SbRLK1 for receptor-like protein kinase from S. bicolor. The putative cytoplasmic domain of SbRLK1 contains all amino acids that are characteristic of protein kinases. The extracellular domain contains five leucine-rich repeats. The mRNA of the SbRLK1 gene accumulated to much higher levels in mesophyll cells than in the bundle sheath and was almost undetectable in roots. This expression pattern indicates that SbRLK1 might be involved in the regulation of specific processes in mesophyll cells. PMID- 10384731 TI - Indole glucosinolate and auxin biosynthesis in Arabidopsis thaliana (L.) Heynh. glucosinolate mutants and the development of clubroot disease. AB - Mutants and wild type plants of Arabidopsis thaliana were analysed for differences in glucosinolate accumulation patterns, indole-3-acetic acid (IAA) biosynthesis and phenotype. A previously identified series of mutants, termed TU, with altered glucosinolate patterns was used in this study. Only the line TU8 was affected in shoot phenotype (shorter stems, altered branching pattern). Synthesis of IAA and metabolism were not much affected in the TU8 mutant during seedling development, although the content of free IAA peaked earlier in TU8 during plant development than in the wild type. Indole glucosinolates and IAA may, however, be involved in the development of clubroot disease caused by the obligate biotrophic fungus Plasmodiophora brassicae since the TU3 line had a lower infection rate than the wild type, and lines TU3 and TU8 showed decreased symptom development. The decline in clubroot formation was accompanied by a reduced number of fungal structures within the root cortex and slower development of the fungus. Indole glucosinolates were lower in infected roots of TU3 and TU8 than in control roots of these lines, whereas in wild-type plants the differences were not as prominent. Free IAA and indole-3-acetonitrile (IAN) were increased in infected roots of the wild type and mutants with normal clubroot symptoms, whereas they were reduced in infected roots of mutants TU3 and TU8. These results indicate a role for indole glucosinolates and IAN/IAA in relation to symptom development in clubroot disease. PMID- 10384733 TI - Laser ablation of gall bladder stones. AB - Study of laser interaction with calculi is presented. A system of Nd-Yag and Ho Yag pulsed lasers were used to produce fluorescence and plasma signals at the stone surface surrounded by saline and bile fluids. Fourth harmonic from Nd-Yag laser was transmitted to the samples by graded UV optical fibres. Gall bladder stones of various compositions were subjected to the high power Ho-Yag laser. Temporal transients and spectral evolution of plasma and fluorescence signals were monitored by a streak camera. A profile of acoustic pressures generated by shock waves was recorded with sensitive hydrophones placed in the surrounding fluids. Ablation threshold, cavitation process and fluorescence dependence on the laser parameters were studied in detail. Potential of stone identification by fluorescence and possible hydrodynamic model for ablation of biological samples is discussed. PMID- 10384734 TI - Spectral studies on the interaction of DNA and Ru(bipy)2(dppx)2+. AB - The interaction of Ru(bipy)2(dppx)2+ (bipy = 2.2'-bipyridine,dppx = 7,8 dimethyldipyrido phenazine) with the calfthymus DNA has been studied with fluorescence and ultraviolet visible absorption spectroscopy. The results of fluorescence quenching and salt effect show that Ru(bipy)2(dppx)2+ intercalate into the double helix of DNA. The ultraviolet visible absorption spectrum of Ru(bipy)2(dppx)2+, calfthymus DNA, and their interaction indicate that Ru(bipy)2(dppx)2+ intercalate into the double helix of DNA via the ligand dppx. PMID- 10384735 TI - Raman spectroscopic studies of CO2 laser-irradiated human dental enamel. AB - While the effects of carbon dioxide (CO2) laser radiation on the physical properties of human dental enamel are well characterized, little is known regarding laser-induced chemical changes. In this study, enamel was exposed to CO2 laser radiation to induce fusion and recrystallization, and the Raman spectra recorded using both dispersive and Fourier-transformed (FT) Raman spectroscopy. Spectra were compared to a heart-treated specimen of hydroxyapatite (HAP) and enamel. Laser irradiation induced chemical changes which differed from those induced by heat treatment. Comparing the Raman spectra of lased enamel to HAP and tricalcium phosphate (TCP), it is evident that CO2 laser irradiation of enamel causes the partial conversion of HAP to TCP. The effect of laser irradiation is not merely a simple local heating effect as previously thought, since simple heating of enamel leads to the formation of both TCP and Ca(OH)2, while laser treatment of enamel results in the formation of TCP but not Ca(OH)2. PMID- 10384736 TI - Acquired 'theory of mind' impairments following stroke. AB - The ability to attribute thoughts and feelings to self and others ('theory of mind') has been hypothesised to have an innate neural basis and a dedicated cognitive mechanism. Evidence in favour of this proposal has come from autism; a brain-based developmental disorder which appears to be characterised by impaired theory of mind, despite sometimes good general reasoning skills/IQ. To date no case of specific acquired theory of mind impairment has been reported. The present study examined theory of mind in adults who had suffered right hemisphere stroke, a group known to show pragmatic and social difficulties. In one study using story materials and two using cartoons, patients' understanding of materials requiring attribution of mental states (e.g. ignorance, false belief) was significantly worse than their understanding of non-mental control materials. Data from healthy elderly subjects, and a small group of left hemisphere patients (who received the tasks in modified form), suggest that this impairment on mental state tasks is not a function of task difficulty. The findings support the notion of a dedicated cognitive system for theory of mind, and suggest a role for the healthy right hemisphere in the attribution of mental states. PMID- 10384737 TI - The scope of teleological thinking in preschool children. AB - These studies explore the scope of young children's teleological tendency to view entities as 'designed for purposes'. One view ('Selective Teleology') argues that teleology is an innate, basic mode of thinking that, throughout development, is selectively applied by children and adults to artifacts and biological properties. An alternative proposal ('Promiscuous Teleology') argues that teleological reasoning derives from children's knowledge of intentionality and is not restricted to any particular category of phenomena until later in development. Two studies explored the predictions of these two hypotheses regarding the scope of children's functional intuitions. Using different methods, both studies found that, unlike adults, pre-schoolers tend to attribute functions to all kinds of objects--clocks, tigers, clouds and their parts. A third study then explored this finding further by examining whether the developmental effect was due to differences in children's and adults' concept of function. It found that both children and adults predominantly view an object's function as the activity it was designed to perform. Possible explanations for the developmental differences found in the first two studies, and implications for notions of a teleological stance are discussed. PMID- 10384738 TI - A case study of an English-Japanese bilingual with monolingual dyslexia. AB - We report the case of AS, a 16 year-old English/Japanese bilingual boy, whose reading/writing difficulties are confined to English only. AS was born in Japan to a highly literate Australian father and English mother, and goes to a Japanese selective senior high school in Japan. His spoken language at home is English. AS's reading in logographic Japanese Kanji and syllabic Kana is equivalent to that of Japanese undergraduates or graduates. In contrast, his performance in various reading and writing tests in English as well as tasks involving phonological processing was very poor, even when compared to his Japanese contemporaries. Yet he has no problem with letter names or letter sounds, and his phoneme categorisation is well within the normal range of English native speakers. In order to account for our data that show a clear dissociation between AS's ability to read English and Japanese, we put forward the 'hypothesis of granularity and transparency'. It is postulated that any language where orthography-to-phonology mapping is transparent, or even opaque, or any language whose orthographic unit representing sound is coarse (i.e. at a whole character or word level) should not produce a high incidence of developmental phonological dyslexia. PMID- 10384739 TI - Specifying the scope of 13-month-olds' expectations for novel words. AB - Recent research has documented that for infants as young as 12-13 months of age, novel words (both count nouns and adjectives) highlight commonalities among objects and, in this way, foster the formation of object categories. The current experiment was designed to capture more precisely the scope of this phenomenon. We asked whether novel words (count nouns; adjectives) are linked specifically to category-based commonalities from the start, or whether they also direct infants' attention to a wider range of commonalities, including property-based commonalities among objects (e.g. color, texture). The results indicate that by 12-13 months, (1) infants have begun to distinguish between novel words presented as count nouns versus. adjectives in fluent, infant-directed speech, and (2) infants expectations for novel words accord with this emerging sensitivity. PMID- 10384740 TI - [Vague symptomatic--the crux of liver diseases.L.Weber, M.D. of Vilshofen explains a plan for liver diagnosis]. PMID- 10384742 TI - [Will blood pressure measuring by the Riva-Rocci and Korotkoff method soon by substituted? Missing oscillometric appliances--technical data kept secret]. PMID- 10384741 TI - [Tuberculosis spectrum displaced. Increasing infection rates by atypical Mycobacteria]. PMID- 10384743 TI - [Errors in blood pressure measuring with modern automated machines. Effects of errors in non-invasive oscillometric blood pressure measuring]. AB - Automatic blood pressure measuring devices with digital display are now in common use, both among lay persons and physicians. The oscillometric method is, however, associated with a number of possible sources of error which, among other things, make comparison with the standard Riva-Rocci method difficult. Methodological errors arise from physiological/anatomical variations: circumference of the forearm and wrist, position of the arteries, structure of the surrounding tissue, arterial diameter and also vasomotor function. The latter represents an appreciable uncertainty factor, in particular with measurements obtained from the finger. Technological sources of error play a role in pronounced hypotension or severe hypertension, since the devices are calibrated only for a range extending from 120 to 180 mmHg. The processing by the device of arrhythmic pulses is also critical. Additional sources of error are in handling and interpretation. Overall, these sources of error are such that not all the various types of device available are equally suitable for use by the patient, and it is necessary to place limitations on the application of such devices and to establish rules for their use. PMID- 10384744 TI - [Practical conducting of blood pressure measuring. Important contributing factors and possibilities of errors]. AB - Studies have shown that both laypersons and doctors and medical staff have insufficient knowledge of the proper handling of blood pressure measurement. The resulting deviations in the measured values are of a diagnostically relevant order of magnitude capable of leading, for example, to an erraneous diagnosis of hypertension. Some of the factors frequently receiving too little attention are, for example, the use of an inappropriate cuff size, failure to ensure that the patient is adequately rested, and inappropriate positioning of the arm during measurement. Attention is drawn to the recommendations of the various hypertension societies. PMID- 10384745 TI - [Asthma-grading amplified. The German Ligue of Respiratory Diseases distinguishes only four stages]. PMID- 10384746 TI - [Family physician and pneumonologist have to cooperate closely. Dietrich Rohde, M.D., Muhlheim, on the new therapy suggestions]. PMID- 10384747 TI - [Quality of life and patient compliance during pain therapy. Multicenter study using Tramundin retard]. PMID- 10384748 TI - [Spondylarthritis. 2: characterization of various forms of spondylarthritis]. PMID- 10384749 TI - [Sexual satisfaction for thE handicapped. "Society is the one to determine handicap". Series: Sex Medicine, 6. Conversation with Inge Plangger, dipl. ped., dipl psych]. PMID- 10384750 TI - [Data search made easy. MEDLINE-Research: paying attention to currency of data. Series: Physician & Computer Literature Research, 3]. PMID- 10384751 TI - [With scalpel and sawdust. Impressions of a historical operating room]. PMID- 10384752 TI - [Emergency aid in osteoporosis--results of the FOSIT study (Fosomax International Trial)]. PMID- 10384753 TI - Effect of ankle dorsiflexion range of motion on rearfoot motion during walking. AB - The purpose of this study was to investigate whether the amount of ankle passive dorsiflexion range of motion influences the pattern of frontal plane rearfoot motion during walking. Three-dimensional motion of the rearfoot was measured in two groups of subjects, those with ankle passive dorsiflexion range of motion less than or equal to 10 degrees, and those with ankle passive dorsiflexion range of motion greater than 15 degrees, while they walked along a 6.1-m walkway. The results indicated that the only statistically significant differences between the two groups were in the time to reinversion of the rearfoot and the time to heel off. Slight-to-moderate limitation of ankle passive dorsiflexion range of motion significantly alters the timing, but not the magnitude, of frontal plane rearfoot motion during walking. PMID- 10384754 TI - Center of pressure and its theoretical relationship to foot pathology. AB - The measurement of center of pressure has been widely used in the evaluation of foot function. This article will describe center of pressure and indicate how it can be used to calculate moments about the joint axes of the foot. Various uses of center of pressure described in the literature will be examined. A model based on the use of the location of center of pressure relative to the location of the subtalar joint axis will be proposed as a theoretical explanation of selected foot pathologies and their treatment. PMID- 10384755 TI - Imaging of the rearfoot. AB - The authors review various pedal conditions affecting the rearfoot, including plantar fasciitis, Achilles tendon pathology, fractures, arthritides, coalitions, and tumors. Various diagnostic imaging modalities such as routine radiography, radionuclide bone scanning, computed tomography, and magnetic resonance imaging are discussed. PMID- 10384756 TI - An update on repair of Achilles tendon rupture. Acute and delayed. AB - Current surgical treatments for Achilles tendon rupture are thoroughly discussed. New repair techniques, such as the use of soft-tissue anchors, are reviewed, as is the use of synthetic mesh to augment the surgical repair. A classification system devised by the author is presented to make it easier to select the appropriate surgical procedure or combination of procedures in delayed rupture repair. Postoperative physical therapy is paramount in the return to preinjury level of activity for these patients. PMID- 10384757 TI - Pleomorphic hyalinized angiectatic tumor of soft parts. AB - Pleomorphic hyalinized angiectatic tumor of soft parts is a recently described neoplasm that most commonly affects the lower extremities. It is locally aggressive but has not been known to metastasize. This article presents a case of a softball-sized tumor on the dorsum of the foot that was identified as pleomorphic hyalinized angiectatic tumor of soft parts. Previously, it would have been misdiagnosed and treated as either a neurilemoma or a malignant fibrous histiocytoma. PMID- 10384758 TI - Effect of podiatric medical care on rates of lower-extremity amputation in a Medicare population. AB - The purpose of this study was to determine whether Medicare patients at risk for lower-extremity amputation due to complications from diabetes, peripheral vascular disease, and/or gangrene who receive the services classified under Level II code M0101 of the Health Care Financing Administration's Common Procedure Coding System (cutting or removal of corns, calluses, and/or trimming of nails, application of skin creams and other hygienic and preventive maintenance care) have lower rates of lower-extremity amputation than those who do not receive such services. Analysis of the data suggests that those at-risk beneficiaries who received these services were nearly four times less likely to experience lower extremity amputation than those who did not receive such services. The study has both methodologic limitations (the study considers only one variable, receipt or nonreceipt of certain types of podiatric medical care, while other variables may affect rates of lower-extremity amputation) and technological limitations (attempts to link the 2 years of per case Medicare Part B data were unsuccessful, limiting the length of the study to 1 year). Further research on this topic is encouraged. PMID- 10384759 TI - Pasteurella multocida and gout in the first metatarsophalangeal joint. PMID- 10384760 TI - Lipoma of the first metatarsophalangeal joint. PMID- 10384761 TI - Lower-extremity gangrene secondary to disseminated intravascular coagulation. PMID- 10384762 TI - Free tissue transfer in the treatment of diabetic foot ulcers. PMID- 10384763 TI - A is for acne. AB - Acne is caused by hormones, androgen, and follicular keratinization, which leads to blocked pores, and P. acnes bacteria, which cause pustule form. Dermatologists report that acne treatments, like the skin of suffers, are clearly getting better. New treatments and the refinements of mainstay ones have changed the face of acne treatment. One advantage of the newer, more effective treatments is that patients now take lower doses of antibiotics for shorter periods of time. PMID- 10384764 TI - Fatigue that doesn't go away. AB - The classic profile of the chronic fatigue syndrome (CFS) patient is a white, middle-age female. Characterized by profound fatigue, CFS often starts with an acute viral infection. While today's medicine provides symptomatic relief, research is offering innovative insights. With this research, is a cure for these patients just around the corner? PMID- 10384765 TI - The impact of auto accidents on medical care. AB - Dual air bags in cars have been standard equipment since 1994. Crumple zones are built into cars to absorb the impact of frontal and rear-end crashes. Virtually all states have mandated seatbelt use laws, and many states, including New Jersey, require motorcyclists to wear a helmet. Yet, more than 3 million Americans were injured in motor vehicle crashes in 1997, and 150,000 to 170,000 of those injuries were serious or extremely serious. PMID- 10384766 TI - Vagal nerve stimulator: a new approach to medically refractory epilepsy. AB - Repetitive vagal nerve stimulation (VNS) is a new, FDA-approved treatment for medically refractory epilepsy. The device is implanted subcutaneously in the left chest and sends intermittent impulses to the left vagus nerve through communicating leads. Twelve patients have been implanted to date. The ages of the patients range from 8 to 36 years and the average followup at this point is five months. Five patients have achieved a greater than 50 percent reduction in seizure frequency with no serious adverse effects. PMID- 10384767 TI - Cancer in the 18th century. A historical interpretation. PMID- 10384768 TI - [The Swiss cardiology foundation: for whom? For what?]. PMID- 10384769 TI - [Treatment of valve diseases]. PMID- 10384770 TI - [Coronary disease: therapeutic gains]. PMID- 10384771 TI - [Endocavitary and endovascular ultrasonography]. PMID- 10384772 TI - [Three-dimensional echocardiography: curiosity or method of the future]. PMID- 10384773 TI - [New indications for cardiac stimulation]. PMID- 10384774 TI - [Treatment of arrhythmias by implantable automatic defibrillators]. PMID- 10384775 TI - [Treatment of arrhythmias by endocavitary ablation]. PMID- 10384776 TI - [Diagnostic and treatment of community-acquired pneumonia in the adult. Recommendations for clinical practice. By the "Practice guidelines for community acquired pneumonia" Work Group]. PMID- 10384778 TI - [When the capital defends its power]. PMID- 10384777 TI - [Following "low threshold": the "optimal minimum" in the therapeutic approach to the difficult patient]. PMID- 10384779 TI - Geochemical and cosmochemical materials. PMID- 10384780 TI - Coatings. PMID- 10384781 TI - Industrial hygiene chemistry: keeping pace with rapid change in the workplace. PMID- 10384782 TI - Air pollution. PMID- 10384783 TI - Water analysis. PMID- 10384784 TI - Pharmaceuticals and related drugs. PMID- 10384785 TI - Forensic science. PMID- 10384786 TI - Environmental analysis. PMID- 10384787 TI - Immunoassays. PMID- 10384788 TI - High-performance liquid chromatography in clinical analysis. PMID- 10384789 TI - Electroanalysis and biosensors. PMID- 10384790 TI - Flame, flameless, and plasma spectroscopy. PMID- 10384791 TI - Infrared spectroscopy. PMID- 10384792 TI - Clinical analyzers. Immunoassays. PMID- 10384793 TI - Clinical analyzers. Advances in automated cell counting. AB - Recently, several hematology analyzers have been developed to improve accuracy and to reduce manual work. The new analyzers use traditional impedance technology or optical detectors alone or in combination. The introduction of argon laser technology to clinical instruments is one new development that may permit integrated flow cytometry parameters leading to a whole new horizon in routine hematologic analyses. PMID- 10384794 TI - Clinical analyzers. Microbiology. PMID- 10384795 TI - Poison center planning for mass gatherings: the Georgia Poison Center experience with the 1996 Centennial Olympic Games. AB - BACKGROUND: The host city for the Olympic Games needs to prepare for "the world" to visit. This article is the first published report describing the impact of an event such as the Olympics on poison center services. PLANNING AND ACTIONS: We evaluated our operations and identified potential new demands. Operational aspects reviewed included: staffing; communication with foreign language callers; knowledge of international drug products; telephone access; procedures for disaster response and recovery; poison treatment protocols; and handling of hazardous material releases. We collaborated with local, state, and federal planners to delineate the poison center role and to develop protocols for use in a poisoning, including a hazardous material's release at a densely populated site. We enhanced our capability to respond to unusual incidents by forming new alliances. Fortunately, no such events occurred; these plans were therefore not tested. CONCLUSIONS: The Georgia Poison Center developed and implemented new capabilities for the Centennial Olympic Games. Immediate access to poison center resources would have facilitated the care of poisoned patients at multiple hospitals if this had become necessary. Communities preparing for mass gatherings should capitalize on the common interests of poison centers, hazardous materials specialists, and public safety agencies. Preparations should emphasize the most likely events, while recognizing the possibility of the unexpected. Planning an integrated response allows the talents and resources of participating agencies to be maximally utilized. PMID- 10384796 TI - Assessment of erythrocyte cholinesterase activity in victims of smoke inhalation. AB - BACKGROUND: The nature of the toxic gases that cause death from smoke inhalation is incompletely understood, and the mechanisms leading to incapacitation remain to be determined. Thermal degradation products of various compounds, including phosphorous-based fire retardants, are suspected capable of impairing human cholinesterase activity. The aim of this study was to measure the erythrocyte cholinesterase activity in victims of smoke inhalation. METHODS: We prospectively measured the erythrocyte cholinesterase activity in blood samples obtained at the scene of residential fires from 49 fire victims. We compared the results with those in an unmatched group of 45 persons with acute drug poisoning. RESULTS: The median (25th-75th percentiles) erythrocyte cholinesterase activity in the 49 fire victims, 1968 IU/L (1660-2276), was significantly lower than in the 45 control subjects 2460 IU/mL (1968-2890), (p = 0.0004). There was no significant difference of the red blood cell counts or plasma protein levels between the 2 groups, while the hematocrit was significantly greater in the fire victims than in the drug-poisoned patients. There was a significant correlation between blood cyanide and carbon monoxide concentrations in the fire victims (r = 0.494, p = 0.002). There was no correlation between erythrocyte cholinesterase activity and either blood cyanide (r = 0.11, p = 0.44) or blood carbon monoxide concentrations (r = 0.04, p = 0.78). CONCLUSIONS: We found a significantly lower level of erythrocyte cholinesterase activity in victims of residential fires, when compared with a convenience sample of hospitalized poisoned patients. Despite the limitations of the study, investigations of the toxic gases potentially responsible for impairment of cholinesterase activity and the clinical significance of this lower enzymatic activity merit further investigation. PMID- 10384797 TI - Prothiofos metabolites in human poisoning. AB - CASE REPORT: A 63-year-old woman was admitted to a local hospital after the ingestion of 40% prothiofos preparation (Tokuthion) 370 mL. Gastric lavage was performed and cathartics, active charcoal, diuretics, atropine sulfate, and pralidoxime were administered. Serum cholinesterase activity was 1.3 IU/L (normal 200-460 IU/L). The patient's consciousness was gradually restored after 4 hours of charcoal hemoperfusion and she was discharged 5 days after admission with no sequelae. METHOD: Plasma and urine prothiofos and metabolites were detected by gas chromatography-flame photometry and gas chromatography-mass spectrometry. Two despropyl metabolites were synthesized for identification and estimation. RESULTS: The main metabolites were identified with authentic prothiofos and methyl esters of synthesized des-S-propyl prothiofos oxon (O-2,4-dichlorophenyl O ethyl phosphate), despropyl prothiofos oxon (O-2,4-dichlorophenyl O-ethyl phospholothiolate), and des-S-propyl prothiofos (O-2,4-dichlorophenyl O-ethyl phosphorothioate). Despropyl prothiofos (O-2,4-dichlorophenyl O-ethyl phosphorodithioate) was also identified in plasma. Large amounts of the hydrolyzed product, 2,4-dichlorophenol and its conjugate were also found. The metabolic pattern of prothiofos in humans appears to be different from that in rats. PMID- 10384798 TI - Acute, fatal, oral chromic acid poisoning. AB - CASE REPORT: We report a 35-year-old woman who developed severe acidosis, massive gastrointestinal hemorrhage, acute renal failure, and hepatic injury following ingestion of chromic acid (50 mL) and died 12 hours after ingestion. Postmortem liver biopsy revealed a fatty degeneration with chromium concentration 3.6 mumol/g. The kidney, with chromium concentration 2.6 mumol/g, had extensive necrosis and ischemic lesions. Erythrocyte chromium was 1903 mumol/L at 3 hours declining to 865 mumol/L at 11 hours. PMID- 10384799 TI - Ferrocyanide ingestion may cause false positives in cyanide determination. AB - CASE REPORT: The authors present a patient who ingested a cyanide containing solution and arrived at the hospital without any clinical evidence of intoxication but an elevated blood cyanide level. The authors explain this discrepancy with the following hypotheses: 1) the patient ingested cyanide as an iron-chelated complex; and 2) the sulfuric acid used in the standard microdiffusion technique released cyanide from its iron-bound state to result in the observed elevated blood cyanide. Through a series of in vitro analyses, the authors demonstrate the following: 1) the ingested solution tested positive for cyanide with the sulfuric acid technique and negative for cyanide with acetic acid; 2) the presence of a ferrous salt in the ingested product by a colorimetric redox titration technique; and 3) release of a small fraction of the total cyanide from ferrocyanide by the sulfuric acid technique. The authors conclude: 1) the patient ingested potassium ferrocyanide; and 2) the strong acid used in the cyanide microdiffusion assay will liberate cyanide that is chelated to iron to yield false positive results. PMID- 10384800 TI - Recovery after Ecstasy intoxication. PMID- 10384801 TI - Ecstasy intoxication and gastric lavage. PMID- 10384802 TI - Organophosphate poisoning: no clear etiological origin. PMID- 10384803 TI - Adolescent gynecology in the office setting. AB - As stated by the Committee on Adolescence of the AAP, All pediatricians who choose to see teenagers should be able to provide counseling about sexual behavior, education on contraceptive methods and prevention of sexually transmitted diseases, and assistance with access to family planning services, preferably in the office or, if necessary, by referral. Expansion of skills and office capabilities to provide routine reproductive health care for adolescents, including pelvic examinations, annual Pap tests, diagnoses of pregnancy, diagnoses and treatment of sexually transmitted diseases, and the prescription of contraceptives, is also encouraged. Pediatricians who have longstanding relationships with their patients, and who are admired and trusted are ideal for providing this care. PMID- 10384804 TI - Menstrual disorders. Amenorrhea. AB - Adolescent patients with amenorrhea often present to primary care providers. A basic understanding of menstrual and pubertal physiology enables clinicians to initiate the clinical evaluation. A thorough history and physical examination focusing on pubertal development indicate the appropriate diagnostic algorithm. Usually, an accurate diagnosis can be obtained quickly. Management includes restoring ovulatory cycles if possible, replacing estrogen when necessary, reassurance, and re-evaluation. PMID- 10384806 TI - Menstrual disorders. Dysfunctional uterine bleeding. AB - Dysfunctional uterine bleeding is defined as abnormal endometrial bleeding without structural pathology. Although, abnormal uterine bleeding may occur in women of all ages, it is a particularly common issue for adolescents. This article reviews the normal menstrual cycle and the abnormal menstrual cycles. Familiarity with the normal menstrual cycle is necessary to identify the possible causes of abnormal bleeding and aids in the diagnosis and treatment of patients with abnormal uterine bleeding. PMID- 10384805 TI - Menstrual disorders in adolescents. Excess androgens and the polycystic ovary syndrome. AB - The fundamental clinical features of PCOS include hirsutism and menstrual irregularities from the time of menarche. Obesity is present in approximately 50% of these patients, some of whom also carry a diagnosis of NIDDM. The biochemical abnormalities associated with the clinical picture include LH hypersecretion, hyperandrogenism, acyclic estrogen production, subnormal SHBG levels, and hyperinsulinemia. Hirsutism usually progresses slowly in patients with PCOS; however, the clinical presentation can resemble virilizing tumors, late-onset CAH, or Cushing syndrome. Virilization or rapidly progressive hirsutism requires immediate investigation to rule out a virilizing tumor. Goals of therapy for teenage patients include decreasing levels of bioavailable androgen, blockade of androgen action at target tissues, stabilization of the endometrium, and reduction of insulin resistance. Although the original description of PCOS by Stein and Leventhal was published in 1935, the cause of PCOS remains unknown. This reason, coupled with the fact that PCOS-related insulin resistance is an important cause of NIDDM in women, has caused this disorder to become one of interest and active investigation. Future research will likely be able to delineate mechanisms behind the defects of carbohydrate metabolism and ascertain large multigeneration kindreds for linkage analyses to identify affected genes. Future studies are also likely to confirm whether young women with PCOS are at increased risk for cardiovascular disease and other long-term health complications. As new pathophysiologic mechanisms are identified, the promise of new therapies arises, including treatments that could potentially reduce the long term incidence of adverse health consequences. PMID- 10384807 TI - Dysmenorrhea and pelvic pain in adolescents. AB - Dysmenorrhea and pelvic pain are common complaints in the adolescent population. Although most cases are primary dysmenorrhea and easily treated with NSAIDs or OCPs, pathologic causes should be considered, especially in cases not responding to standard medical management. Endometriosis is the most common finding in teenagers who do not respond to this regimen, but mullerian anomalies and musculoskeletal causes must also be considered. PMID- 10384808 TI - Evaluation of adnexal masses in adolescents. AB - The detection of adnexal masses in adolescents is worrisome to patients, their families, and physicians. Reassurance can be given that the vast majority of these lesions are benign. Furthermore, a significant fraction of benign masses are functional ovarian cysts, most of which resolve spontaneously and never need surgery. Imaging is critical in determining the management of these patients. Sonography is the preferred first-line diagnostic tool. When surgery is necessary, physicians must recognize the importance of conserving the ovaries and uterus to avoid the loss of reproductive and endocrine function. PMID- 10384809 TI - Vulvar disorders in adolescent patients. AB - This article describes the most commonly encountered vulvar disorders in adolescents and refers to some less common entities. The categories are based upon appearance of the vulvar lesions: solid, cystic, ulcerative and infectious. PMID- 10384810 TI - Breast disease in adolescents and young women. AB - Breast lesions, including mastalgia; benign proliferative changes; and benign masses, including fibroadenomas, are common in adolescents and young adult women. Breast cancer is rare in women less than 20 years old and uncommon in women less than 30 years old. Discrete masses in women less than 30 years old that do not feel suspicious on examination can be observed for 1 or 2 months. If they persist, than an FNA can differentiate those that are benign and can be observed versus those that need either an excisional biopsy or definitive surgery. Mammography has little role in the diagnosis of women less than 30 years old except in those individuals with highly suspicious lesions on examination. PMID- 10384812 TI - Phytoestrogens and bone. AB - Practically all plant foods contain small amounts of the diverse phytoestrogen molecules that have the potential to improve health. Phytoestrogens, especially the soy-derived isoflavones, are receiving great scrutiny as food supplements for the purposes of both enhancing the health of tissues and preventing several common diseases, such as cardiovascular diseases, cancers of reproductive tissues and osteoporosis. Investigations of isoflavones, in particular, have recently become more prominent because of their oestrogenic activities. These actions may be as either partial oestrogen agonists or anti-oestrogens (inhibitors of natural oestrogen activity). For example, the isoflavones of soy, mainly genistein and daidzein, have been shown by at least three different laboratories to conserve bone in ovariectomized rodent models, and they probably have similar conservatory effects in higher mammalian species. Nevertheless, the only positive effects of phytoestrogens on bone observed so far in post-menopausal women have been small and limited to the lumbar vertebrae. Additional information on human studies currently in progress is needed before the efficacy of these preparations in human subjects is known. PMID- 10384811 TI - Gynecologic care of medically complicated adolescents. AB - Gynecologic care of adolescents presents a challenge under the best of circumstances, but when the patient has significant medical processes that interact with the process of puberty, the care of these patients may become extremely difficult. A review of the more common medical illnesses of adolescents and the interaction on the events of puberty and normal menstrual function is presented with emphasis on contraception and future fertility. Although many of the contraceptive options present a possible increased risk to these patients, it must be kept in mind that these adolescent patients will develop emotionally and become sexually active at some point in their lives, and the potential risk of the resultant pregnancy must be weighed carefully. The various options of management for gynecologic problems are discussed. PMID- 10384813 TI - Phytoestrogens and breast cancer. AB - Phytoestrogens are paradoxical. Because of their structural similarity to the physiological oestrogens, they have been assumed to increase the risk of breast cancer. However, nations where the largest amounts of phytoestrogens are consumed in the diet have the lowest incidence of and rate of death from breast cancer. Although these epidemiological observations do not prove that phytoestrogens have anti-cancer properties, many preclinical experiments support this concept. Some indicate that early life exposure to phytoestrogens may be critical for breast cancer prevention. Clinical studies to define the effect of phytoestrogens on breast cancer recurrence are underway. The recent discovery of a second class of oestrogen receptors, with a differential distribution among the tissues, may enable an explanation of the phytoestrogen paradox. These receptors have opened a way of utilizing phytoestrogens in the treatment of oestrogen-sensitive chronic diseases such as atherosclerosis and osteoporosis. PMID- 10384814 TI - Phytoestrogens and the menopause. AB - For most women, the menopause presents two sets of problems. First, most notice unpleasant symptoms such as hot flushes and vaginal dryness, but second, there are long-term sequelae arising from oestrogen deficiency. The main long-term problems are an increased risk of bone loss and cardiovascular disease. This chapter will focus on the role of phytoestrogens in alleviating menopausal symptoms. Studies to date would suggest that phytoestrogenic products may help around two-thirds of women to cope with menopausal symptoms such as hot flushes, but there is little evidence that these products will help with vaginal dryness. It seems probable that these products lower cholesterol and therefore cardiovascular risk; however, it is important that women who use such products to alleviate menopausal symptoms have a bone density performed every 2 or 3 years to assess their risk of osteoporosis. PMID- 10384815 TI - Phytoestrogens and coronary heart disease. AB - While there have been ample studies of a cross-cultural nature and experimental evaluations establishing the cardioprotective effect of soy protein, efforts to clarify the proportion of those benefits related to its phytoestrogen content are relatively recent. In most cases, the general approach to evaluating the role of soy's phytoestrogens has been to compare the cardiovascular benefits of isolated soy protein with a comparable soy protein isolate that has been alcohol extracted. Based on that approach, soy phytoestrogens appear to lower low-density lipoprotein concentrations while increasing plasma concentrations of the high density lipoproteins. Particularly noteworthy with respect to the high-density lipoprotein effects are the increases in apolipoprotein A-1. Phytoestrogens may also prevent the oxidation of lipoprotein particles. The soy phytoestrogens favourably influence coronary artery reactivity. They also inhibit the progression of atherosclerosis in the coronary, iliac and common and internal carotid arteries. The cardiovascular benefits of soy phytoestrogens appear to be equal for males and females. PMID- 10384816 TI - Epidemiology of phytoestrogens. AB - Epidemiological studies have revealed that high levels of lignans and isoflavonoids are frequently associated with low breast, prostate and colon cancer risk, as well as a low risk of coronary heart disease. These compounds seem to be cancer protective and/or are biomarkers of a 'healthy' diet. All soy protein products consumed by Asian populations have high concentrations of isoflavonoids. In other countries, such as Finland and Sweden, the lignan levels are higher in populations with the lowest risk because of a high consumption of whole-grain rye bread, berries and some vegetables. There is a strong association between fibre intake per kilogram body weight and lignan concentrations in body fluids. Breast cancer has been found to be associated with low lignan levels in the USA, Finland, Sweden and Australia. With regard to prostate and colon cancer, as well as coronary heart disease, the epidemiological data related to phytoestrogens are still very limited. PMID- 10384817 TI - Phytoestrogens and diseases of the prostate gland. AB - Both benign hyperplasia (BPH) and cancer of the prostate are manifest in men beyond the age of 50. Approximately 50% of men greater than 50 years of age will suffer from the symptoms associated with BPH, especially from bladder outlet obstruction. With the ever-increasing proportion of the population over 65 years of age worldwide, BPH is becoming an important medical problem as the world moves into the next millennium. Cancer of the prostate is the second most commonly diagnosed cancer after skin cancer in the male population of the United States, and the second most common cause of death from cancer after that of the lung. Overall, around the world the incidence of carcinoma of the prostate is increasing annually by 2-3%. Both race and geographical location have a profound influence of the prevalence of prostate cancer worldwide. Black men in the USA have the highest incidence, while the incidence is much lower in Asian men from China, Japan and Thailand. Although the prostate gland is androgen-dependent, it is now recognized that the biological actions of endocrine-related factors, such as androgens, oestrogens, glucocorticoids and certain dietary and environmental factors, are mediated within the gland by various growth regulatory factors. The growth regulatory factors such as epidermal growth factor (EGF), keratinocyte growth factors (KGF), fibroblast growth factors (FGFs) and insulin-like growth factors II and I are mitogenic and directly stimulate cell proliferation under the modulating influence of steroid hormones. Steroids are therefore essential but not directly responsible for cell proliferation. Certain plant compounds such as isoflavonoids, flavonoids and lignans have been proposed as cancer protective compounds in populations with low incidences of prostate diseases. In particular, soya contains the isoflavone genistein, a compound with many properties which could influence both endocrine and growth factor signalling pathways. PMID- 10384818 TI - Phytoestrogens and inhibition of angiogenesis. AB - The consumption of a plant-based diet can prevent the development and progression of chronic diseases associated with extensive neovascularization, including the progression and growth of solid malignant tumours. We have previously shown that the plant-derived isoflavonoid genistein is a potent inhibitor of cell proliferation and in vitro angiogenesis. Moreover, the concentration of genistein in the urine of subjects consuming a plant-based diet is 30-fold higher than that in subjects consuming a traditional Western diet. We have also reported that certain structurally related flavonoids are more potent inhibitors than genistein. Indeed, 3-hydroxyflavone, 3',4'-dihydroxyflavone, 2',3' dihydroxyflavone, fisetin, apigenin and luteolin inhibit the proliferation of normal and tumour cells as well as in vitro angiogenesis at half-maximal concentrations in the lower micromolar range. The wide distribution of isoflavonoids and flavonoids in the plant kingdom, together with their anti angiogenic and anti-mitotic properties, suggest that these phytoestrogens may contribute to the preventive effect of a plant-based diet on chronic diseases, including solid tumours. PMID- 10384820 TI - Experimental studies on lignans and cancer. AB - Mammalian lignans are produced from plant precursors such as secoisolariciresinol diglycoside (SDG) and matairesinol via the action of bacteria in the human or animal colon. While precursors are found in many plant foods, flaxseed is the richest source of SDG and was therefore used as a model to determine the anti cancer effects of lignans. This paper reviews the experimental studies in animals and humans demonstrating the anti-cancer effects of flaxseed and its SDG as well as other studies relevant to the clinical use of lignans, such as those on their food sources, bio-availability and safety. PMID- 10384819 TI - Reproductive actions of phytoestrogens. AB - This chapter reviews the reproductive actions of phytoestrogens, comparing mechanisms of action, dose-response relationships, and human exposures. Although a wide range of biochemical actions have been reported for phytoestrogens, in vitro tests suggest that phytoestrogens may be more likely to act through receptor-mediated mechanisms than through enzyme inhibition. Epithelial cell proliferation in the reproductive tract and anestrus are well-documented actions of isoflavonoids in experimental studies of animals. However, thus far, soy-based diets have generally failed to produce epithelial proliferation in ovariectomized rats and monkeys or menopausal women, and clinical studies have produced mixed evidence for effects of soy isoflavones on the human menstrual cycle or post menopausal gonadotropin secretion. There has been considerable interest in the use of phytoestrogens as oestrogen replacement therapy in menopausal women. Reported results of initial clinical trials have been mixed, and it is unclear whether isoflavones in presently advised doses can substantially reduce menopausal symptoms. Some recent trials with oral isoflavone supplements report reductions in hot flushes, vaginal dryness, and breast pain. There is also limited clinical evidence for protective actions of isoflavones in mammary cancer. Like other oestrogenic substances, the isoflavonoids are effective differentiating agents in rodent models of development. The consequences of these actions for humans is of interest due to the high concentrations of isoflavonoids in some infant formulae. Thus, it is likely that some humans may experience greater exposure to phytoestrogens in infancy than in any other lifestage. At the time of writing, no ill effects of such exposure have been reported. PMID- 10384821 TI - Soyfoods, isoflavones and risk of colonic cancer: a review of the in vitro and in vivo data. AB - Soy foods and soybean components have received considerable attention of late for their potential role in reducing cancer risk. Although the relationship between soy intake and the risk of breast and prostate cancer has been the focus of most interest, the relationship between soy intake and other cancers, including colorectal cancer, has also been studied. Several anti-carcinogens have been identified in soybeans, but most enthusiasm for the potential anti-cancer effects of soy undoubtedly stems from work involving soybean isoflavones. Isoflavones have a limited distribution in nature, and, for practical purposes, soyfoods are the only nutritionally relevant dietary source of these phytochemicals. Isoflavones are weak oestrogens but possess other potentially important biological attributes independent of their ability to bind to the oestrogen receptor. The isoflavone genistein inhibits the growth of most types of hormone dependent and hormone-independent cancer cells in vitro, including colonic cancer cells. Several mechanisms for the in vitro anti-cancer effects of genistein have been proposed, including effects on signal transduction. A number of epidemiological studies, primarily of Asian origin, have examined the relationship between soy intake and the risk of colorectal cancer. Although these studies provide little support for a protective effect of soy, concerns have been raised about the completeness of the soy intake data, since soy was not the focus of these studies and most of this research was conducted prior to the recent interest in the anti-cancer effects of soy. The effect of soy/isoflavone intake has also been studied in rodents, but again these data are conflicting and provide only modest support for a protective effect. Although the relationship between soy intake and colonic cancer risk is certainly worthy of further investigation, there is, at the moment, very limited support for soy exerting a protective effect against this type of cancer. PMID- 10384822 TI - Phytoestrogen content in foods. AB - Plants abound in essential phytochemicals produced for their various vital functions. The same compounds seem also to be crucial for human health and disease. Recent human epidemiological and laboratory animal and cell studies on cancer and heart disease have highlighted the phytoestrogens--naturally occurring principles that share with steroidal oestrogens an ability to activate oestrogen receptors. The best known non-steroidal phytoestrogens include the isoflavones daidzein, genistein, formononetin and biochanin A, the coumestan coumestrol, and the lignans secoisolariciresinol and matairesinol. Acknowledging the potentially chemoprotective role of these non-nutrients, we have quantified all biologically important isoflavonoids and lignans in cereals, oilseeds and nuts, legumes, vegetables, fruits, berries and beverages such as tea, coffee and wine. In this chapter, we present a review of our studies on staple plant foods, indicating that plants contain, besides a wide range of chemicals with a number of biological properties, biologically active phytoestrogens--precursors of hormone like compounds found in mammalian systems. PMID- 10384823 TI - Unstable angina. Introduction. PMID- 10384824 TI - The health care burden of unstable angina. AB - Worldwide, UA represents a significant allocation of resources. UA represents a syndrome where not only do many therapies exist, but considerable clinical trial evidence has accumulated. Universal application of effective practice patterns is warranted if we are to successfully reduce the burden of UA. Economic analyses cannot resolve many of the underlying societal issues that affect decision making. Often, the acceptability of an economic burden is dependent on the willingness of both individuals and society to pay. In an interesting study, Chestnut et al evaluated the willingness of 50 patients to pay for avoiding a worsening of their angina symptoms. On average, the patients were willing to pay between $210 and $499 to avoid four to eight additional angina episodes each month. The "rule of rescue" suggests that society is often willing to pay large sums of money to save those in extreme need, such as the 55-year-old man rushed to the emergency department clutching his chest. Only recently has attention been paid to how much this disease entity costs us. Whereas the 1980s and 1990s saw a focus on costs, the next century will increasingly focus on value--obtaining the best health outcome for the dollars spent. Debate has shifted, at least in part, from purely financial costs to medical effectiveness and outcomes management. Continuing assessments of value of interventions and application of evidence based-management strategies permit rational selection of therapy and allow us best to bear the burden of UA. PMID- 10384825 TI - Histopathology of plaque rupture. AB - Plaque disruption occurs during the development of atherosclerotic lesions. During certain circumstances it may result in thrombosis and subsequent development of acute coronary syndromes. Several characteristics of the plaque appear to be associated with plaque disruption, including a large lipid rich core, superficial plaque inflammation, and a thin fibrous cap. The importance of these and other plaque components are discussed in this article. PMID- 10384826 TI - Plaque disruption and thrombosis. Potential role of inflammation and infection. AB - Considerable data from in vitro and in vivo studies of vascular biology, together with indirect evidence from clinical trials of lipid-lowering or modifying and lifestyle or risk factor modifying interventions, provide strong support for the concept that disruption of atherosclerotic plaque and subsequent thrombosis is a key precipitant of potentially lethal, acute coronary syndromes. Certain characteristics of plaques, including the size and composition of the lipid core, the structure and composition of the fibrous cap, and the presence of a local inflammatory process, predispose the plaque to disruption. Stresses resulting from biomechanical and hemodynamic forces acting on plaques may then trigger disruption, releasing the thrombogenic contents of the lipid core. Alterations in endothelial function may also contribute to vulnerability of plaque rupture and thrombosis. Therefore, interventions aimed at decreasing plaque vulnerability to disruption--all based on the concept of plaque stabilization--may reduce the risk of acute coronary syndromes. Although not yet rigorously validated in humans, plaque stabilization may prove to be an important clinical strategy for preventing the lethal consequences of coronary atherosclerosis. PMID- 10384827 TI - Thrombosis and coagulation abnormalities in the acute coronary syndromes. AB - The acute coronary syndromes, that include unstable angina, acute myocardial infarction, and many cases of sudden cardiac death, exact a considerable price on society in terms of mortality, morbidity, and health care costs. The coronary atherosclerotic lesion is often an indolent and progressive entity that can destabilize causing an acute syndrome with or without warning. The majority of acute coronary syndromes result from events such as rupture or disruption of the atherosclerotic plaque with intracoronary thrombosis and ischemia of the distal myocardium as a result. Advances in our understanding of the process underlying the acute coronary syndromes has allowed for the identification of targets and rational therapeutic strategies for the prevention and treatment of these syndromes. Many of these therapeutic strategies involve the reversal of prethrombotic forces that often coexist with coronary atherosclerosis. Even with recent advances in our approach to atherosclerosis, intracoronary thrombosis, and the resulting acute coronary syndromes, an unacceptably high event rate persists after these syndromes. Further advances in the prevention and treatment of coronary atherosclerosis and its thrombotic complications depends on a more thorough understanding of the biology of the atherosclerotic plaque and the factors which influence its stability. PMID- 10384828 TI - Characterization of the unstable lesion by angiography, angioscopy, and intravascular ultrasound. AB - Acute coronary syndromes are a spectrum of clinical presentations with various pathophysiologic substrates. As such, there is not one single type of lesion responsible for stable or unstable angina, acute myocardial infarction, and sudden cardiac death. Most of the information regarding the characteristics of culprit lesions derives from histopathologic studies, whether postmortem or from atherectomy samples, and from studies using angiography, angioscopy, and intravascular ultrasound. Characterization of the unstable coronary lesion is key to understanding the pathophysiology of coronary artery disease, this knowledge will allow clinicians to individualize treatment according to specific lesion types, and more importantly, will lead to strategies to identify atherosclerotic lesions in their early stages and implement preventive therapies. PMID- 10384829 TI - Evaluating the chest pain patient. Scope of the problem. AB - With an understanding of the pathophysiology of ACS and an increasing number of early therapeutic options, there has been a shift in focus from ruling-out MI to identifying and stratifying risk in all patients with potential ACS. The presenting symptoms and ECG still remain the cornerstone of immediate diagnosis and triage. Through the application of new technologies, such as the cardiac troponins, and a reassessment of techniques, such as perfusion imaging and echocardiography, the clinician has an increasing selection of methods to rapidly assess chest pain of potential ischemic etiology. Coinciding with the evaluation of technology has been the development of the concept of the CPU and associated rapid diagnostic protocols. These protocols, whether they utilize the assistance of mathematic predictive instruments or represent simple triage schemes, form the backbone of a system to improve the care of patients with ACS in the current milieu of cost containment. PMID- 10384830 TI - Anticoagulant therapy in unstable angina. AB - The goal of anticoagulant therapy in unstable angina is to prevent progression of a subocclusive coronary thrombus to complete occlusion of the coronary artery, thereby preventing myocardial infarction and death. Although these have been many advances in therapy with anticoagulants, considerable morbidity and mortality remains. Also, although combination therapy with potent novel anticoagulants and antiplatelet agents may be an alternative strategy, this needs to be balanced against the risks of hemorrhagic complications. More precise and biologically relevant methods of monitoring anticoagulant effect, along with appropriately modified doses given in combination offers promise. PMID- 10384831 TI - Antiplatelet therapy for treatment of acute coronary syndromes. AB - Acute coronary syndromes and the postpercutaneous coronary intervention state share the common feature of atherosclerotic plaque disruption and subsequent intraluminal thrombus formation. In most cases, vascular patency is maintained but partial occlusion causes myocardial ischemia and can either progress to complete occlusion or result in distal embolization with subsequent small vessel obstruction, the core section of an intraarterial thrombus is platelet-rich and can serve as a nidus for further thrombosis. Aspirin, by virtue of its anticycloxygenase activity inhibits platelet activation and aggregation to a mild degree. Clinically, aspirin has been shown to reduce the rates of myocardial infarction in patients with acute coronary syndromes and to reduce the number of ischemic complications which follow coronary angioplasty. More potent inhibitors of platelet aggregation antagonize the interaction between the platelet surface protein GP IIb-IIIa and fibrinogen. The result is profound inhibition of platelet aggregation. Three intravenous antagonists of platelet GP IIb-IIIa are clinically available and a fourth is under phase III study. When used in addition to aspirin therapy, these agents have been shown to produce further reductions in either peri-interventional infarctions or in recurrent myocardial infarctions in patients with acute coronary syndromes. PMID- 10384832 TI - "Traditional" medical therapy for unstable angina. How important? How to use? AB - Unstable angina comprises a heterogeneous population of patients who present with a wide spectrum of underlying pathophysiology. The traditional treatment of these patients is based on both evidenced-based medicine as well as clinical experience. Despite the large population of patients admitted with this diagnosis, the scientific literature regarding its treatment is scarce. Therefore, the management of patients with unstable angina relies heavily on the clinical skills of the physician. One of the most important steps in this process involves risk stratification, especially in the current environment of cost containment. Those patients who are at low risk for adverse outcomes can be treated and evaluated safely as outpatients. Patients at high or moderate risk, however, should be treated intensively as inpatients. Although there appear to be many new promising therapies for unstable angina on the horizon, the traditional therapies still have a place. The use of aspirin in this population is well supported by the literature and appears to have a positive effect on mortality and cardiovascular events. The other traditional therapies, however, are not as well supported by the literature. They do appear to benefit the patient in terms of reducing symptoms, but their effects on reducing mortality and cardiovascular events are not clear. Therefore, the goal of medical therapy in this patient population should be to stabilize them so that they can proceed with an appropriate risk stratification procedure as soon as possible. This is especially true with performing coronary angiography or interventions because the risk of procedural complications is higher in patients with unstable angina and ongoing symptoms. PMID- 10384833 TI - Early use of coronary angiography and intervention. AB - In this article we have outlined the current rationale and role of invasive management in ACS. For the majority of patients with ACS, who are either at high risk or unstable, invasive management is a critical element in breaking the sequence of recurrent ischemia leading to early cardiac events (Fig. 11). Secular trends in the care of cardiovascular patients predict even more sophisticated, invasive methods of treating coronary occlusion in the future. A futurist's view on this subject may envision the following type of scenario. A patient with prior CAD experiences persistent chest pain and notifies the emergency medical system. The paramedics arrive, and perform a rapid fingerstick cardiac biomarker panel and ECG. The results are interpreted by an emergency physician via a telecommunication system, and the patient is determined to be at high risk. He or she is triaged to a center capable of angioplasty and bypass surgery. On the way to the hospital, the patient is treated with aspirin, IV heparin, and an IV glycoprotein IIb/IIIa inhibitor. The patient undergoes triage angiography within 1 hour of hospital arrival, culprit lesion(s) are identified, and a revascularization plan is made--setting a critical pathway that is definitive. This vision is not far off on the horizon. We anticipate additional clinical trial results will help form the decision points in this optimal treatment scenario, which for a large proportion of patients will involve invasive management. PMID- 10384834 TI - Aggressive versus conservative therapy in unstable angina. AB - In patients with unstable angina, non-Q-wave, and Q-wave myocardial infarction, atherosclerotic plaque rupture leads to a variable amount of platelet adhesion and aggregation, vasoconstriction, and partially or totally occlusive thrombus formation. This article focuses on the role of aggressive (routine angiography and revascularization) versus conservative (maximal medical therapy, with catheterization and revascularization reserved for those with spontaneous or provable ischemia) management of the patient with unstable angina. PMID- 10384835 TI - Interesting cases from the University of Texas Medical Branch. AB - This article discusses the cases for four patients with unstable angina. The first case is an example of the "high-risk" patient with widespread ECG changes, heart failure, and enzymatic elevations during an episode of chest pain. The second patient illustrates an unusual cause of unstable angina in a young women. The third patient had a large thrombus visible on angiography and management strategies for dealing with intracoronary thrombus are discussed. The final patient had an extensive past cardiac history with two prior coronary artery bypass operations and we discuss the recent advances made in the treatment of degenerative vein graft disease. PMID- 10384837 TI - Effective head and neck cancer management: a consensus document. PMID- 10384836 TI - Management of high-risk subsets in unstable angina. AB - This article focuses on the optimal treatment of postinfarction, refractory, or recurrent angina based on the results of recent clinical trials. Many of our recommendations hold true for the general management of unstable angina, but special considerations for the high-risk subsets are emphasized. Specifically, we discuss acute medical management and suggest that an early aggressive strategy that leads to early coronary angiography with the goal of revascularization when feasible best serves this subset. A special emphasis on the emerging role of glycoprotein IIb-IIIa antagonists is made because the important role of platelets in coronary thrombosis has dominated recent views on the pathophysiology of unstable angina. PMID- 10384838 TI - Ginkgo biloba for tinnitus: a review. PMID- 10384839 TI - Disintegration of porous polyethylene prostheses. AB - A Plastipore (porous polyethylene) Total Ossicular Replacement Prosthesis gave an excellent initial hearing result which was maintained for 14 years. Hearing then began to deteriorate and revision surgery showed disintegration of the prosthesis and a defect in the stapes footplate. Histological examination confirmed previous findings in porous polyethylene with multinucleated foreign body giant cells and breakdown of the material. PMID- 10384840 TI - The BAHA HC200/300 in comparison with conventional bone conduction hearing aids. AB - A retrospective study was performed on 89 patients from a consecutive series who received a BAHA HC200/300 after having previously used conventional bone conduction hearing aids. The patients' performance with the BAHA HC 200/300 was compared to their performance with conventional bone conduction hearing aids. The patients were divided into two groups, depending on the time of implantation (before or after May 1992). The patients in group 1 (long-term users) were asked to fill in a questionnaire, the same one as they had filled in at the initial BAHA fitting more than 5 years previously. The answers were compared to their original opinions and difference scores were calculated. The long-term clinical results from group 1 are also presented. Although they are encouraging, the patients' opinion about the BAHA deteriorated somewhat over time. The audiometric results of group 2 were highly comparable with those of group 1. This confirms the positive results with the BAHA found in previous studies. PMID- 10384841 TI - The implant-site split-skin graft technique for the bone-anchored hearing aid. AB - We describe the technique of implant-site split-skin grafting for the bone anchored hearing aid (BAHA). Twenty-five patients have undergone this procedure (20 adults and five children) since 1993 with a minimum follow-up of 1 year. Fifteen adults were operated upon as single stage surgery, all other cases (including all children) were performed in two stages. In four patients (16%) significant early graft inflammation was encountered which settled with outpatient treatment. In one the abutment had to be temporarily removed to allow the graft to settle. All patients now have a stable graft site. This surgical technique is straightforward and a separate graft donor site is avoided. It would appear this technique results in a stable BAHA graft site with low associated morbidity. PMID- 10384842 TI - Effect of conductive hearing loss on the vestibulo-collic reflex. AB - The vestibulo-collic reflex represents a promising test for evaluating the integrity of otolith function. We have investigated the threshold of this response in a group of normal subjects, and the effect of a conductive hearing loss. A positive response was recorded in 31 of 32 normal subjects. The threshold of the vestibulo-collic reflex varied from 80 to 97 dBHL in these subjects with a 95% response rate at a threshold at 96 dBHL. A total of 23 ears with a conductive hearing loss in 17 patients were also investigated. The average conductive hearing loss (at 0.5, 1, 2 and 4 kHz) ranged from 8.75 to 40 dBHL (average 24.46 dBHL). A positive response was recorded in only two ears. Therefore, the vestibulo-collic reflex has a high stimulus threshold which is dependant on reliable transmission of the click stimulus to the inner ear thus limiting is clinical use. PMID- 10384843 TI - The smell map: is there a commonality of odour perception? AB - The normal perception of odour quality is poorly understood, so formulating meaningful tests of olfaction is difficult. While tests of odour discrimination and odour detection threshold have helped quantify olfactory dysfunction, there are not yet predictive relationships between sensitivity to particular odours and particular forms of olfactory dysfunction. Using 11 commonly encountered odours, 20 normosmics performed similarity ratings of odour pairs. Multidimensional scaling, a standard behavioural sciences data analysis method, was used to explore the perceptual relationships between the odours based on their pair-wise similarity ratings. Smell maps were created for each individual as was a common or archetypal map which indicated a commonality in individuals' odour perception, far greater than chance alone (P < 10(-6)). A preliminary analysis of four hyposmics suggests that they do not conform to the normosmic archetype. Future studies assessing the relationship between odours in the archetype should improve the selection of odours to be included in tests of odour discrimination. PMID- 10384844 TI - Xenogeneic ossicular implants: an experimental study of heterotopic, demineralized, lyophilized, porcine implants in the guinea-pig. AB - This study was done to compare the outcome of porcine ossicular implants in the middle ear and the subcutaneous dorsal region of the guinea-pig to those of allo implants implanted in parallel in the dorsal region. The implants were heteropic, xenogeneic, demineralized (HCl), lyophilized and sterilized. The evaluation was histological (light microscopy and scanning electron microscopy) and immunological (immunofluorescence staining). Fifty-four guinea-pigs were implanted in the middle ear and 14 of them were also implanted subcutaneously in the dorsal region with xeno-implants and allo-implants. The middle ear implants were found to be constantly reossified and coated with normal mucosa with only a minimal immune reaction. In contrast, the dorsal xeno-implants were found to be the target of mononucleic infiltration, fibrous encapsulation and an influx of immunoglobulins resulting in segregation. The corresponding allo-implants were found to be partially reoccupied and reossified. These findings highlight the value of HCl demineralization in the induction of non-species-specific Bone Morphogenetic Protein and the failure of attempts at immuno-despecification. It appears that the fate of the implant depends less on its antigenic load than on the site of implantation. In this regard the middle ear is apparently very advantageous. The very good short-term tolerance and recovery observed in the middle ear xeno-implant suggest that these implants offer sufficiently good results to warrant clinical testing. PMID- 10384845 TI - Informed consent: British otolaryngologists surveyed. AB - A repeat postal questionnaire of British otolaryngologists has been carried out to assess changes in their practice of obtaining informed consent. The previously reported high level of good practice is maintained. Currently informed consent is more often obtained in outpatients or the preadmission clinic than it was in 1991. Consultants and specialist registrars are more involved in this process. Although there is little evidence given to support a persistent increase in a defensive approach towards gaining informed consent, there is some change in the reporting of surgical complications that may reflect an increased awareness of the concept of 'material risk'. PMID- 10384846 TI - Day case paediatric tonsillectomy: a review of three years experience in a dedicated day case unit. AB - Tonsillectomy is one of the most common surgical procedures performed in children. Day case tonsillectomy is common in many parts of the USA but remains controversial in the UK. Day case tonsillectomy in children has been performed in a dedicated paediatric day care unit at the May day Hospital since 1994. The results for the years 1995-1997 were reviewed. In these years 928 true day case tonsillectomies were performed. Reactionary haemorrhage occurred in 0.97% (nine children) but only three children required a return to theatre. All bleeding occurred within the standard observation period. There were no deaths. The overall effective day case rate was 95.7%, only 31 children needing unplanned admission, mainly for postoperative vomiting. The results suggest that day case tonsillectomy can be safely and successfully performed, with a dedicated paediatric day case unit and a favourable population geography. PMID- 10384847 TI - Admission rates, early readmission rates and patient acceptability of 142 cases of day case septoplasty. AB - Day case surgery should be confined to those procedures where less than 3% of patients require admission. The aim of this study was to establish the admission rates, early readmission rates and patient acceptability of 142 consecutive cases of day case septoplasty. Data acquisition was by retrospective postal questionnaire. One hundred and fifty-three patients were studied and data was acquired on 142. Ninety per cent (128/142) of patients had operations on afternoon lists. Admissions were 7/142 (5%), the early readmission rate (within 24 h) was 0% and 25/142 (17%) of patients felt they would rather have stayed in hospital for the first night after surgery. The conclusion of this work is that day case septoplasty is an acceptable practice in appropriately selected patients who are operated upon in the morning and when the technique described here is applied. An acceptably small proportion of planned day cases may require admission. PMID- 10384848 TI - External ear resonance in patients with tympanic membrane perforations. AB - This study investigated the effects of a tympanic membrane perforation on the external ear resonance. Measurements of external ear resonance using a probe-tube microphone system were performed in 14 patients who had medium to large unilateral tympanic membrane perforations. The contralateral normal ears of these 14 patients served as control. The results showed that there were no significant differences in the peak frequency, peak amplitude and peak sharpness between perforated and normal ears. However, intersubject variability in the resonant frequency was greater in the perforated group. In addition, the resonance curves of these two groups were substantially different. In 10 out of the 14 patients in the perforated group, the resonance curves showed 2-3 prominent peaks separated by valleys of about 10 dB reduced gain. In addition, in 11 out of 14 perforated ears, reduced responses (3.8 dB in average) occurred consistently in the lower frequency region (0.3-2 kHz). Clinically, the abnormal external ear resonance and the larger intersubject variation must be taken into consideration in fitting hearing aids for this group of patients. PMID- 10384849 TI - Sensorineural hearing loss in chronic otitis media. AB - Although many studies have demonstrated an association between chronic otitis media (COM) and sensorineural hearing loss (SNHL), there still remains disagreement about the relationship. A retrospective study was conducted to examine the relationship between sensorineural hearing loss and chronic otitis media. Forty-one patients met the following criteria: unilateral COM and no history of head injury, meningitis or previous otological surgery. The differences in preoperative bone conduction threshold between diseased and control (contralateral normal) ear were statistically significant (P < 0.01) and varied from 5.24 to 9.02 dB across the frequency range. The effect of duration of disease on the degree of SNHL was also analysed but no correlation was found. The presence of cholesteatoma and/or ossicular erosion was not associated with a significantly increased risk of sensorineural hearing loss. PMID- 10384850 TI - Sonotubometry findings in children at high risk from middle ear effusion. AB - The functioning of the eustachian tube has an important role to play in the development of middle ear disease. It would be useful if a clinical test could assist in the identification of eustachian tube dysfunction, particularly if this is shown to be an indicator of persistent middle ear effusion. The aim of this study was to compare the results of sonotubometry using the MMS-10 instrument in children at high risk from middle ear effusion with a group of normal subjects. Forty-one subjects (age range 5-6 years) were allocated to one of two groups (experimental group, 21 subjects; control group 20 subjects) based on a questionnaire designed to identify subjects at high risk from middle ear effusion. The test protocol allowed each subject to swallow three times for each of two pure-tones (7 and 8 kHz) delivered by the nasal probe. Sonotubometry indicated opening of the eustachian tube on swallowing in around 80% of subjects. The incidence of positive findings varied greatly amongst subjects across both groups. In the control group, the mean increase in sound pressure level on swallowing was 11.5 dB (+/- 4.3) and 9.8 dB (+/- 2.5) for 7 and 8 kHz, respectively. The corresponding means for duration were 118 ms (+/- 47.9) and 137 ms (+/- 61.8). Sonotubometry failed to demonstrate a difference between the two groups of subjects. Hence, the clinical application of sonotubometry to identify subjects at high risk from middle ear effusion is not supported. PMID- 10384851 TI - A randomised clinical trial of antiseptic nasal carrier cream and silver nitrate cautery in the treatment of recurrent anterior epistaxis. AB - Sixty-four consecutive patients with a history of recurrent epistaxis were randomly assigned in the outpatient clinic to receive treatment with either Naseptin antiseptic nasal carrier cream alone (Group A) or a combination of Naseptin cream and silver nitrate cautery (Group B). Results were available on 50 patients, 22 in Group A and 28 in Group B. Twenty patients (91%) in Group A and 25 patients (89%) in Group B demonstrated improvement in their symptoms. There was no statistically significant difference in outcome between the two treatment arms (P = 0.7569). On comparing the different age groups (under and over 16 years) in the two treatment arms, once again there was no statistically significant difference in the treatment outcome (P = 1.000). In conclusion, silver nitrate cautery offers no added advantage to the management of simple epistaxis in both children and adults. PMID- 10384852 TI - Thiobarbituric acid reactive substances in patients with laryngeal cancer. AB - The purpose of this study was to measure the level of thiobarbituric acid reactive substances (TBARS), which are products of lipid peroxidation, as a reflection of oxidative status in 30 patients with laryngeal cancer. The TBARS were measured 1 day before surgery and 3 weeks after laryngectomy compared to 30 normal control subjects by the thiobarbituric acid (TBA) test using spectrophotometry. The TBARS levels were significantly elevated in patients with laryngeal cancer compared to the controls (18.4 +/- 3.13 mumol/l versus 7.75 +/- 1.9 mumol/l; P < 0.05). There were no differences in the TBARS levels in relation to T staging, N staging, or degree of differentiation. Among patients with laryngeal cancer smokers had slightly higher TBARS levels. Comparison of the paired observations of laryngeal cancer larynx before and after operation revealed a significant rise in the TBARS levels after laryngectomy. Tissue damage resulting from surgery is the most probable explanation for this rise. The state of altered lipid peroxidation detected in our patients might be related to a deficient antioxidant status. The role of antioxidants in the prevention of laryngeal cancer among high risk patients is worth studying. PMID- 10384853 TI - Computed tomography evaluation of the inner ear as a diagnostic, counselling and management strategy in patients with congenital sensorineural hearing impairment. AB - The value of computed tomography (CT) of the petrous bone in the investigation of congenital sensorineural hearing impairment has been questioned. We have conducted a study to establish the usefulness of CT of the temporal bone in the evaluation and management of a consecutive series of unselected adolescent patients with congenital sensorineural hearing impairment of greater than 50 dB HL. Seventy-one patients (142 ears) were identified and images reviewed to establish the incidence of inner ear malformations. Fifteen ears were found to be abnormal in eight patients (seven bilateral and one unilateral abnormality). Three patients had Mondini abnormalities and one of these also had dilatation of the lateral semicircular canals. There were five patients with dilatation of the vestibular aqueduct. One patient had a unilateral dysplasia of the middle and external ear. A variety of incidental intracranial abnormalities were also discovered. We conclude that CT does have a valuable role in the management of SNHI. PMID- 10384854 TI - The Epworth Sleepiness Scale: can it be used for sleep apnoea screening among snorers? AB - Snoring is a common disorder and may lead to the development of Obstructive Sleep Apnoea (OSA) with its associated hazards. Differentiation of patients with OSA from patients with simple snoring is crucial to the ENT surgeon before selecting treatment. This study aimed to assess the reliability of the Epworth Sleepiness Scale (ESS) to screen for OSA among snorers. Forty-six patients referred for treatment of snoring were studied. Each patient completed the ESS questionnaire and subsequently underwent a hospital sleep study. The ESS scores did not correlate with the apnoea/hypopnoea indices calculated from the sleep studies (correlation coefficient 0.12). The lack of correlation is mainly because simple snorers can also suffer from excessive daytime sleepiness, due to an unclear mechanism. The ESS is a useful questionnaire for assessing disability as a result of snoring but it is of no value in distinguishing simple snorers from patients with OSA. PMID- 10384855 TI - Assessment of mucociliary transport in patients with chronic mucoid rhinitis. AB - Chronic rhinitis is the manifestation of a heterogeneous group of disease entities and often proves difficult to manage successfully. We present the investigations of the mucociliary system in 40 patients with mucoid rhinorrhoea as their principal symptom of whom 20 had pan respiratory disease. The saccharin clearance time (SCT) was measured and classified as normal if it was below 20 min. Objective measurement of clearance was made using 99mTechnetium-labelled human serum albumin (99mTc-HSA). We have standardized our method using a micrometer syringe driver to produce a droplet of consistent size (droplet size, 0.01 ml, SD 0.0002 ml) that reduces the dose of radiation. The movement of the droplet was measured over 20 min (RLT). The mean, maximum rate and percentage moved were calculated. Patients were divided into those who had chest disease (20) and those without and a chi 2-test was performed for the mean RLT time between the two groups. There was a strong correlation between mean and maximum rates (r = 0.91). One patient has a normal SCT and normal RLT. Patients with chest disease had a significantly lower mean RLT (P > 0.01). Assuming that RLT is the standard investigation, six patients were normal but had an abnormal SCT, this is a false positive error of 15%. The false negative error was 4/40 (10%). The association between sinus and chest disease with abnormal mucociliary clearance is stressed. PMID- 10384856 TI - Interactions between oral contraceptives and antifungals/antibacterials. Is contraceptive failure the result? AB - The effectiveness of oral contraceptives may be impaired by concomitant treatment with antimicrobials. This may occur because of reductions in plasma concentrations of ethinylestradiol by the induction of hepatic metabolism, as for rifampicin (rifampin) and possibly griseofulvin, or in a small percentage of women because of interference with enterohepatic recirculation. There are no scientific data to support the anecdotal evidence that the concomitant use of combined oral contraceptives and antimicrobials reduces contraceptive efficacy in the majority of women. It has been postulated that there is a subset of women in whom the enterohepatic recirculation of ethinylestradiol plays an important role. In these women the action of an antimicrobial may reduce the efficacy of oral contraceptives by interfering with this mechanism. Studies that have quantitatively examined these effects may have failed to include women from this subset because of the small numbers involved in the studies. On the other hand, there are no good prospective studies comparing contraceptive failure rates between compliant women who use combined oral contraceptives with and without antimicrobials. All women using combined oral contraceptives should be informed of the very low level of risk of interactions with antimicrobials (probably about 1%) and that it is not possible to identify who may be at risk. Women concerned about this low level of risk should be given information about the use of barrier methods or avoidance of intercourse during the first 7 days of concomitant antimicrobial therapy and for 7 subsequent days. Women who have had previous contraceptive failures or developed breakthrough bleeding during concomitant antimicrobial use should be strongly advised to follow these precautions, as they may be part of the subset of women at higher risk of contraceptive failure. PMID- 10384857 TI - Pharmacokinetics of alendronate. AB - Alendronate (alendronic acid; 4-amino-1-hydroxybutylidene bisphosphonate) has demonstrated effectiveness orally in the treatment and prevention of postmenopausal osteoporosis, corticosteroid-induced osteoporosis and Paget's disease of the bone. Its primary mechanism of action involves the inhibition of osteoclastic bone resorption. The pharmacokinetics and pharmacodynamics of alendronate must be interpreted in the context of its unique properties, which include targeting to the skeleton and incorporation into the skeletal matrix. Preclinically, alendronate is not metabolised in animals and is cleared from the plasma by uptake into bone and elimination via renal excretion. Although soon after administration the drug distributes widely in the body, this transient state is rapidly followed by a nonsaturable redistribution to skeletal tissues. Oral bioavailability is about 0.9 to 1.8%, and food markedly inhibits oral absorption. Removal of the drug from bone reflects the underlying rate of turnover of the skeleton. Renal clearance appears to involve both glomerular filtration and a specialised secretory pathway. Clinically, the pharmacokinetics of alendronate have been characterised almost exclusively based on urinary excretion data because of the extremely low concentrations achieved after oral administration. After intravenous administration of radiolabelled alendronate to women, no metabolites of the drug were detectable and urinary excretion was the sole means of elimination. About 40 to 60% of the dose is retained for a long time in the body, presumably in the skeleton, with no evidence of saturation or influence of one intravenous dose on the pharmacokinetics of subsequent doses. The oral bioavailability of alendronate in the fasted state is about 0.7%, with no significant difference between men and women. Absorption and disposition appear independent of dose. Food substantially reduces the bioavailability of oral alendronate; otherwise, no substantive drug interactions have been identified. The pharmacokinetic properties of alendronate are evident pharmacodynamically. Alendronate treatment results in an early and dose-dependent inhibition of skeletal resorption, which can be followed clinically with biochemical markers, and which ultimately reaches a plateau and is slowly reversible upon discontinuation of the drug. These findings reflect the uptake of the drug into bone, where it exerts its pharmacological activity, and a time course that results from the long residence time in the skeleton. The net result is that alendronate corrects the underlying imbalance in skeletal turnover characteristic of several disease states. In women with postmenopausal osteoporosis, for example, alendronate treatment results in increases in bone mass and a reduction in fracture incidence, including at the hip. PMID- 10384861 TI - Cutaneous leishmaniasis (CL) is an important public health problem in many countries. PMID- 10384858 TI - Clinical pharmacology of new histamine H1 receptor antagonists. AB - The recently introduced H1 receptor antagonists ebastine, fexofenadine and mizolastine, and the relatively new H1 antagonists acrivastine, astemizole, azelastine, cetirizine, levocabastine and loratadine, are diverse in terms of chemical structure and clinical pharmacology, although they have similar efficacy in the treatment of patients with allergic disorders. Acrivastine is characterised by a short terminal elimination half-life (t1/2 beta) [1.7 hours] and an 8-hour duration of action. Astemizole and its metabolites, in contrast, have relatively long terminal t1/2 beta values; astemizole has a duration of action of at least 24 hours and is characterised by a long-lasting residual action after a short course of treatment. Azelastine, which has a half-life of approximately 22 hours, is primarily administered intranasally although an oral dosage formulation is used in some countries. Cetirizine is eliminated largely unchanged in the urine, has a terminal t1/2 beta of approximately 7 hours and a duration of action of at least 24 hours. Ebastine is extensively and rapidly metabolised to its active metabolite; carebastine, has a half-life of approximately 15 hours and duration of action of at least 24 hours. Fexofenadine, eliminated largely unchanged in the faeces and urine, has a terminal t1/2 beta of approximately 14 hours and duration of action of 24 hours, making it suitable for once or twice daily administration. Levocabastine has a terminal t1/2 beta of 35 to 40 hours regardless of the route of administration, but is only available as a topical application administered intranasally or ophthalmically in patients with allergic rhinoconjunctivitis. Loratadine is rapidly metabolised to an active metabolite descarboethoxyloratadine and has a 24-hour duration of action. Mizolastine has a terminal t1/2 beta of approximately 13 hours and duration of action of at least 24 hours. Most orally administered new H1 receptor antagonists are well absorbed and appear to be extensively distributed into body tissues; many are highly protein-bound. Most of the new H1 antagonists do not accumulate in tissues during repeated administration and have a residual action of less than 3 days after a short course has been completed. Tachyphylaxis, or loss of peripheral H1 receptor blocking activity during regular daily use, has not been found for any new H1 antagonist. Understanding the pharmacokinetics and pharmacodynamics of these new H1 antagonists provides the objective basis for selection of an appropriate dose and dosage interval and the rationale for modification in the dosage regimen that may be needed in special populations, including elderly patients, and those with hepatic dysfunction or renal dysfunction. The studies cited in this review provide the scientific foundation for using the new H1 antagonists with optimal effectiveness and safety. PMID- 10384860 TI - The efficiency concept in pharmacodynamics. AB - The classic approach to describe the pharmacological response to a drug is to analyse its concentration-effect relationship, using a variety of possible models such as maximum effect (Emax) models or sigmoid Emax models. The aim of this review is to discuss an alternative way of describing the pharmacological effect in terms of effect per unit of drug concentration, instead of simple effect. This variable is called efficiency, analogous with concepts used in other fields. The pharmacodynamic model for efficiency is derived from the sigmoid Emax model and is dependent on the same parameters. Since the sigmoid Emax model incorporates 'the law of diminishing returns', requiring ever higher concentrations to increase the effect by a given percentage, efficiency is bound to decrease with increasing concentrations. However, as a mathematical consequence of its derivation from the sigmoid Emax model, efficiency also has a maximum value, which can be expressed as a function of the slope factor (s) and drug concentration associated with half the maximum effect (C50), provided that the slope factor is greater than 1. The efficiency concept is potentially applicable to all drugs and particularly useful for those that follow concentration-effect relationships according to Emax or sigmoid Emax models. Most experience has been obtained with loop diuretics, especially with furosemide (frusemide). Slow administration of furosemide, leading to slow excretion of the drug, has been shown, in many studies, to significantly increase the total diuretic effect per amount of drug recovered in urine. In this review, some examples of the applicability of the efficiency concept to other drugs, such as antibacterials, opioids and antineoplastics, are discussed. In addition to pharmacodynamically varying efficiency, other saturable processes, such as the formation of active metabolites and saturable transport, may form a basis for the application of the efficiency concept. The efficiency of a drug dosage may also be influenced by tolerance and counter-regulation produced by the drug. All these factors contribute to schedule dependency. It is concluded that the shape of the time course of drug presentation to its site of action is an independent determinant of overall response. The possibility of adjusting the drug input profile to maximise therapeutic effect per dose and to separate cumulated therapeutic from cumulated adverse effects should be considered in designing administration schedules and in drug development. PMID- 10384862 TI - Cutaneous leishmaniasis: a historical perspective. PMID- 10384859 TI - Pharmacokinetics and pharmacodynamics of the nitroimidazole antimicrobials. AB - Metronidazole, the prototype nitroimidazole antimicrobial, was originally introduced to treat Trichomonas vaginalis, but is now used for the treatment of anaerobic and protozoal infections. The nitroimidazoles are bactericidal through toxic metabolites which cause DNA strand breakage. Resistance, both clinical and microbiological, has been described only rarely. Metronidazole given orally is absorbed almost completely, with bioavailability > 90% for tablets; absorption is unaffected by infection. Rectal and intravaginal absorption are 67 to 82%, and 20 to 56%, of the dose, respectively. Metronidazole is distributed widely and has low protein binding (< 20%). The volume of distribution at steady state in adults is 0.51 to 1.1 L/kg. Metronidazole reaches 60 to 100% of plasma concentrations in most tissues studied, including the central nervous system, but does not reach high concentrations in placental tissue. Metronidazole is extensively metabolised by the liver to 5 metabolites. The hydroxy metabolite has biological activity of 30 to 65% and a longer elimination half-life than the parent compound. The majority of metronidazole and its metabolites are excreted in urine and faeces, with less than 12% excreted unchanged in urine. The pharmacokinetics of metronidazole are unaffected by acute or chronic renal failure, haemodialysis, continuous ambulatory peritoneal dialysis, age, pregnancy or enteric disease. Renal dysfunction reduces the elimination of metronidazole metabolites; however, no toxicity has been documented and dosage alterations are unnecessary. Liver disease leads to a decreased clearance of metronidazole and dosage reduction is recommended. Recent pharmacodynamic studies of metronidazole have demonstrated activity for 12 to 24 hours after administration of metronidazole 1 g. The post antibiotic effect of metronidazole extends beyond 3 hours after the concentration falls below the minimum inhibitory concentration (MIC). The concentration dependent bactericidal activity, prolonged half-life and sustained activity in plasma support the clinical evaluation of higher doses of metronidazole given less frequently. Metronidazole-containing regimens for Helicobacter pylori in combination with proton pump inhibitors demonstrate higher success rates than antimicrobial regimens alone. The pharmacokinetics of metronidazole in gastric fluid appear contradictory to these results, since omeprazole reduces peak drug concentration and area under the concentration-time curve for metronidazole and its hydroxy metabolite; however, concentrations remain above the MIC. Other members of this class include tinidazole, ornidazole and secnidazole. They are also well absorbed and distributed after oral administration. Their only distinguishing features are prolonged half-lives compared with metronidazole. The choice of nitroimidazole may be influenced by the longer administration intervals possible with other members of this class; however, metronidazole remains the predominant antimicrobial for anaerobic and protozoal infections. PMID- 10384863 TI - Epidemiology of cutaneous leishmaniasis. PMID- 10384864 TI - Cutaneous leishmaniasis. The parasite. PMID- 10384865 TI - Leishmania virulence and genetic heterogeneity. PMID- 10384866 TI - Reservoir hosts of cutaneous leishmaniasis. PMID- 10384867 TI - The biology and control of phlebotomine sand flies. PMID- 10384868 TI - Cutaneous leishmaniasis: clinical features and diagnosis. PMID- 10384869 TI - Histologic diagnosis of cutaneous leishmaniasis. PMID- 10384870 TI - Treatment of cutaneous leishmaniasis: retrospectives and advances for the 21st century. PMID- 10384871 TI - Global control and Leishmania HIV co-infection. PMID- 10384872 TI - Cutaneous leishmaniasis: strategies for prevention. PMID- 10384873 TI - Immune response to Leishmania infection in human skin. PMID- 10384874 TI - Recent trends in vaccine development and immunization. PMID- 10384875 TI - Neurons induce GFAP gene promoter of cultured astrocytes from transgenic mice. AB - In order to investigate the influence of neuron-glia interaction on astrocyte differentiation, we used a transgenic mouse bearing part of the gene promoter of the astrocytic maturation marker GFAP linked to the beta-galactosidase (beta-gal) reporter gene. Addition of embryonic cerebral hemisphere (CH) neurons to transgenic CH astrocyte monolayers increased by 50-60% beta-gal positive cell number. Such event was dependent on the brain regional origin of the neurons and was followed by an arrest of astrocytes from the cell cycle and induction of glial differentiation. Time-course assays demonstrated that maximum effect was observed after 24 h of coculture. Addition of conditioned medium (CM) derived from CH neurons also increased beta-gal positive CH astrocytic cell number. However, such CM had no effect on midbrain and cerebellum astroglia. Together, these data suggest that neurons secrete brain region-specific soluble factors which induce GFAP gene promoter, as measured by beta-gal expression, thus suggesting that neuron-glia interaction might induce the astrocytic differentiation program. PMID- 10384876 TI - Dendritic calcium transients in the leech giant glial cell in situ. AB - Glial cells have been shown to respond to neuronal activity with changes in the membrane potential and the intracellular Ca2+ concentration. In order to get closer to glial structures associated with neuronal synapses, we have now looked at Ca2+ signalling in the glial processes ("glial dendrites") in response to neurotransmitters and neuronal activity. Single giant glial cells in situ of isolated ganglia of the leech Hirudo medicinalis were filled iontophoretically with the Ca(2+)-sensitive dyes Oregon green 488 BAPTA-1 or Fluo-3. Relative Ca(2+)-dependent fluorescence changes in response to bath and focal application of the ionotropic glutamate receptor agonist kainate (50 microM) and of 5 hydroxytryptamine (5-HT, 100 microM) were recorded in glial dendrites, using confocal laser scanning microscopy. The amplitudes of the [Ca2+]i transients in the dendritic processes were 2-4 times larger than those recorded in the cell body. Electrical stimulation of a nerve root (20 Hz for 15 s) elicited [Ca2+]i transients in glial dendrites (n = 32) that were reduced by the ionotropic glutamate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; n = 14). The results demonstrate that neuronal activity can evoke [Ca2+]i transients not only in glial cell bodies but also in glial dendrites, where these transients display regional variation. This may reflect local release of neurotransmitters like glutamate and 5-hydroxytryptamine and/or regional differences in the density of glial receptors. PMID- 10384877 TI - Potentiation of ATP calcium responses by A2B receptor stimulation and other signals coupled to Gs proteins in type-1 cerebellar astrocytes. AB - We have studied the interaction between P1 and P2 purinoceptors in purified type 1 astrocyte cultures from postnatal days 7-8 rat cerebella using single cell microfluorimetry with fura-2. The stimulation of astrocytes with ATP elicits rapid [Ca2+]i transients showing an EC50 value of 7.9 +/- 0.3 microM. Costimulation of type-1 astrocytes with adenosine and ineffective ATP concentrations (0.1 or 1 microM) evoked [Ca2+]i transients that correspond to 60% of the maximal ATP response. NECA (5'-N-ethylcarboxamidoadenosine) was the only agonist that mimicked the adenosine effect and showed an EC50 value of 0.17 +/- 0.01 microM. This value was identical to that obtained for the cAMP production stimulation, indicating that A2B receptors coupled to adenylate cyclase activation were involved. The presence of A2B adenosine receptors was also confirmed by immocytochemistry experiments. When astrocytes were costimulated with isoproterenol and ineffective ATP concentrations similar [Ca2+]i transients were observed. The treatment of astrocytes with cholera toxin potentiated ATP calcium signals, lowering the EC50 value for ATP to 1.5 +/- 0.2 microM. However, the pretreatment of cells with forskolin or a permeable cAMP analogue had no effect on ATP calcium responses. These results indicated that the potentiation mechanism was elicited before the adenylate cyclase activation. We could conclude that in type-1 astrocytes, the activation of A2B adenosine receptors or other signals positively coupled to adenylate cyclase stimulation strongly potentiate metabotropic calcium responses to ATP. The potentiation was parallel but independent on cAMP accumulation suggesting the involvement of beta gamma subunits released after Gs stimulation. PMID- 10384878 TI - GABAA receptor agonists modulate K+ currents in adult hippocampal glial cells in situ. AB - Glial cells are known for their role in development and expression of GABA receptors. However, there seems to be a lack of in situ studies characterizing GABA receptor expression and function in glial cells from early development to adulthood. Consequently, we examined GABA receptor expression on rat hippocampal glial cells in both neonatal and adult slices using the whole-cell patch-clamp technique. Glial cells in adult and neonatal slices exhibit responses to muscimol (1 mM; GABAA), but not baclofen (1 mM; GABAB), demonstrating that receptor electrophysiology remains qualitatively similar in glial cells throughout development. Adult muscimol current densities however, do show a decrease in size to approximately 36% of the neonatal response. Muscimol responses were found to be sensitive to bicuculline, suggesting that they are mediated by GABAA receptors. In addition to receptor currents, muscimol causes a concomitant long term blockade of outward K+ currents in glial cells of both neonatal and adult slices. Comparisons of percentage peak blockade in adult and neonatal glial cells show no significant difference. However, when comparing average absolute conductance blockade, we see that adult glial cells display a significantly smaller response than neonatal and cultured astrocytes. Therefore, although the percentage blockade of outward currents remains consistent throughout development, neonatal glial cells display a larger physiological effect. Thus, it can be concluded that, although the complex GABA response in glial cells is affected by development, the receptor current and secondary blockade are a basic mechanism for neuronal-glial interaction throughout life. PMID- 10384879 TI - Tumor necrosis factor-alpha regulation of the Id gene family in astrocytes and microglia during CNS inflammatory injury. AB - The inhibitors of DNA binding (Id) gene family is highly expressed during embryogenesis and throughout adulthood in the rat central nervous system (CNS). In vitro studies suggest that the Id gene family is involved in the regulation of cell proliferation and differentiation. Recently, Id gene expression was shown to be expressed in immature and mature astrocytes during development and upregulated in reactive astrocytes after spinal cord injury. These results suggest that the Id gene family may play an important role in regulating astrocyte development and reactivity; however, the factors regulating Id expression in astrocytes remain undefined. Tumor necrosis factor-alpha (TNF alpha), a proinflammatory cytokine, is thought to play a crucial role in astrocyte/microglia activation after injury to the CNS. To determine if TNF alpha plays a role in Id gene expression, we exogenously administered TNF alpha into developing postnatal rats. We report that TNF alpha injections resulted in a rapid and transient increase in both cell number and mRNA expression for Id2 and Id3 when compared to levels observed in noninjected or control-injected animals. Id1 mRNA levels were also upregulated after TNF alpha treatment, but to a lesser degree. Significant increases in TNF alpha-induced Id2 and Id3 mRNA were observed in the ventricular/subventricular zone, cingulum and corpus callosum. TNF alpha also increased Id2 mRNA expression in the caudate putamen and hippocampus at the injection site. Id2 and Id3 mRNA+ cells were identified as GFAP+ and S100 alpha + astrocytes as well as ED1+ microglia. This is the first report to show TNF-alpha-induced modulation of the Id gene family and suggests that Id may be involved in the formation of reactive astrocytes and activated microglia in the rodent brain. These results suggest a putative role for the Id family in the molecular mechanisms regulating cellular responsiveness to TNF alpha and CNS inflammation. PMID- 10384880 TI - Mice lacking NT-3, and its receptor TrkC, exhibit profound deficiencies in CNS glial cells. AB - Neurotrophin-3 (NT-3) and its receptor TrkC are known to be important for neuronal survival. More recently, NT-3 has been implicated as playing a role in oligodendrocyte (OL) proliferation and survival in vitro. Examination of NT-3 and TrkC knockout mice revealed a reduction in NT-3-dependent neurons. To date, no study has examined alterations in glial cell populations in these knockout mice. In this report, we demonstrate a decline in OL progenitor cell numbers within the CNS of NT-3 and TrkC knockout mice. We also observed that immature and mature OL specific markers were attenuated in the NT-3 and TrkC knockout animals. Deficiencies in other CNS glial cells, including astrocytes and ameboid microglia, were also observed. The subventricular zone (SVZ), a highly proliferative region for progenitor glial cells, was reduced in size. Furthermore, a nuclear-specific stain revealed a decline in the numbers of pyknotic nuclei in and around the SVZ of the knockout mice. These data will support an in vivo NT-3-dependent mechanism for the normal development of CNS glial cells. PMID- 10384881 TI - Identification and localization of Ca(2+)-activated K+ channels in rat sciatic nerve. AB - To understand the physiology of Schwann cells and myelinated nerve, we have been engaged in identifying K+ channels in sciatic nerve and determining their subcellular localization. In the present study, we examined the slo family of Ca(2+)-activated K+ channels, a class of channel that had not previously been identified in myelinated nerve. We have determined that these channels are indeed expressed in peripheral nerve, and have cloned rat homologues of slo that are more than 95% identical to the murine slo. We found that sciatic nerve RNA contained numerous alternatively spliced variants of the slo homologue, as has been seen in other tissues. We raised a polyclonal antibody against a peptide from the carboxyl terminal of the channels. Immunocytochemistry revealed that the channel proteins are in Schwann cells and are associated with canaliculi that run along the outer surface of the cells. They are also relatively concentrated near the node of Ranvier in the Schwann cell outer membrane. This staining pattern is quite similar to what we previously reported for the voltage-dependent K+ channel Kv 1.5. We did not observe staining of axons or connective tissue in the nerve and so it seems likely that most or all of the splicing variants are located in the Schwann cells. The localization of these channels also suggests that they may participate in maintaining the resting potential of the Schwann cells during K+ buffering. PMID- 10384882 TI - Edg-2 in myelin-forming cells: isoforms, genomic mapping, and exclusion in Charcot-Marie-Tooth disease. AB - Edg-2 is an heptahelical receptor whose spatio-temporal distribution during rat brain development is consistent with a role in the control of myelination. We have now identified two splice variants of Edg-2 mRNA in rat brain that encode two receptor isoforms differing by a stretch of 18 amino acids in the NH2 terminal extracellular tail of the receptor. Prenatally (i.e., before oligodendrocyte myelination), the two variants detected by selective in situ hybridization are equally abundant, vary in parallel, and remain restricted to proliferative zones in the brain. Postnatally, the long isoform becomes predominant in myelinating structures, where its abundance increases sharply during the period of myelination. In the adult human brain, only the long variant was detected, while in situ hybridization showed it selectively expressed in the white matter and in clusters of cells showing features of oligodendrocytes of the temporal cerebral cortex. Consequently, the human Edg-2 gene was studied to assess its possible contribution in inherited neuropathies. The coding sequence was found to be contained in three exons and to map to chromosome 9q31.3-32 by using radiation hybrid panel and Yeast-Artificial Chromosomes. Two intragenic bi allelic polymorphisms and a rare mutation were identified. As a first application to molecular genetic studies, they were used to exclude the Edg-2 gene in six families with phenotype of demyelinating Charcot-Marie-Tooth disease of unknown origin. PMID- 10384883 TI - Glial responses during evoked behaviors in the leech. AB - Glial cells can respond with membrane potential changes during electrically stimulated neuronal activity (Kuffler, Proc R Soc Lond B 168:1-21, 1967; Orkand, Oxford University Press, 1995). Their role in contributing to, or controlling, neural circuits underlying behaviors, however, is completely unknown. We have used semi-intact preparations of the leech Hirudo medicinalis, where behaviors can be elicited and monitored (Kristan et al., J Neurobiol 27:380-389, 1995), to record membrane responses of identified glial cells during whole-body shortening and during fictive swimming. Giant glial cells are located in the neuropil of segmental ganglia, where neuronal axons and dendrites establish numerous synaptic contacts (Coggeshall and Fawcett, J Neurophysiol 27:229-289, 1964). We report here that these glial cells hyperpolarize when the whole-body-shortening response is evoked but not during fictive swimming. To our knowledge, this is the first report that associates a specific behavior with glial cell responses. PMID- 10384884 TI - [Androgenetic alopecia. Hair loss effectively stopped with 1 mg Finasterid]. PMID- 10384885 TI - [1. Janssen-Cilag Forum, Congress of Dermatology in New Orleans 1999. Current and future status of dermatology]. PMID- 10384886 TI - Characterization of eae+ Escherichia coli isolated from healthy and diarrheic calves. AB - Strains of Escherichia coli from 101 healthy and 114 diarrheic calves were screened by PCR for the eae (intimin) gene and Shiga toxin genes (stx). Each eae+ and eae/stx+ strain was examined for antimicrobial susceptibility, enterohemolysin activity, and the somatic O antigen was determined. An immunoassay was used to detect Shiga toxin antigens for the eae/stx+ E. coli. Significantly more (p = 0.005) of the healthy calves carried eae+ and eae/stx+ E. coli in their feces when compared to strains from diarrheic calves. Moreover, Shiga toxin antigens were detected significantly more (p = 0.001) often among the eae/stx+ strains from healthy calves when compared to eae/stx+ strains from diarrheic calves. However, significantly more (p = 0.001) of the eae+ and eae/stx+ strains from diarrheic calves were resistant to at least one of the antimicrobials tested, and the strains from diarrheic calves had a significantly (p = 0.05) higher rate of antimicrobial resistance to at least two different antimicrobial classes. No significant difference (p> or =0.05) was detected among the eae+ and eae/stx+ strains from healthy and diarrheic calves for enterohemolysin production. Serogroups O-negative, O5, O26, and O111 were predominate among both healthy and diarrheic calves. PMID- 10384887 TI - Prevalence and characteristics of necrotoxigenic Escherichia coli (NTEC) strains isolated from diarrhoeic dairy calves. AB - Fecal samples from 246, 1-90-days old diarrhoeic dairy calves in 72 herds were screened for the presence of cytotoxic necrotizing factors (CNF)-producing Escherichia coli (NTEC). NTEC were detected by tissue culture assays and PCR in 39 (15.8%) of the diarrheic calves, and the majority of these animals (34 of 39, ca. 87.2%) were infected by NTEC producing CNF2. Calves were grouped according to their age (1-7 days, 8-14 days, 15-21 days, 22-30 days and 31-90 days) and analyses of prevalence were done by the Mantel-Haenzsel chi2-test for trend. A significant age-associated increase in the prevalence of NTEC producing CNF2 (p<0.0001) was found. Eighty-one (8.4%) of the 958 E. coli isolates from the 246 diarrheic calves were positive for CNF in the tissue culture assays. These strains were analyzed by PCR and this technique showed that three (3.7%) strains were CNF1-positive and 75 (92.6%) were CNF2-positive. Moreover, three of the strains positive in the tissue culture assays were negative by PCR. These strains were subsequently assayed in several biological tests (rabbit skin test, mouse intraperitoneal test and mouse footpad test) which showed that they were really NTEC, probably producing CNF2, but with some different properties to classical strains producing CNF2. NTEC strains producing CNF2 belonged to different serogroups (O2, O7, O9, O14, O15, O41, O43, O45, O55, O76, O86, O88, O109, O115, O123, O128, O153 and O159) than strains producing CNF1 (O11 and O32) or PCR negative strains (O111). Moreover, a strong association between CNF2 and F17 fimbriae was found (78.6% of CNF2-positive strains were F17-positive, whereas only 22.9% of CNF2-negative strains were F17-positive). PMID- 10384888 TI - Molecular typing of Staphylococcus aureus of bovine origin by polymorphisms of the coagulase gene. AB - Mastitis caused by Staphylococcus aureus is a disease of major economic importance to the dairy industry. Transmission occurs during milking, with chronically-infected cows acting as the major reservoirs of infection. PCR coagulase gene typing of 151 S. aureus isolates from seven farms generated only six PCR types, with 110 (73%) isolates assigned to PCR type 1 and 23 (15.2%) isolates assigned to type 2. PCR type 1 was the predominant type on five of the seven farms, including farms in geographically separated regions of Victoria, while type 2 predominated on two farms. With the exception of the 41 isolates from one farm, all isolates were resistant to penicillin, but susceptible to other antibiotics that are routinely used to manage mastitis in dairy cattle. Nine of 11 cows with chronic S. aureus infection showed evidence of persistence of a single PCR type for periods of up to 9 months. Two different PCR types of S. aureus were isolated from the other two cows over the same period. PMID- 10384889 TI - A Salmonella abortusovis inactivated vaccine protects mice from abortion after challenge. AB - Two types of Salmonella abortusovis vaccines were prepared, one with aluminium hydroxide (vaccine A) and the other with water in oil (vaccine B) adjuvants. They were compared in a pregnant mouse model, aiming at protecting them from abortions after challenge with a virulent strain of S. abortusovis. The protection for vaccine A was from 74% to 77.6% and that for vaccine B from 71% to 79.6%. Abortions occurred 5-10 days post challenge and S. abortusovis was isolated from all aborted fetuses and from the liver and the spleen of their mothers at the end of the experiment (18 days post challenge). The presence of salmonella in the liver and the spleen of vaccinated non-pregnant but challenged mice was studied in a separate experiment. The bacterium was isolated from one out of 12 vaccinated mice 6 days post challenge as well as from the six controls. PMID- 10384890 TI - Comparative sequence analysis of classical swine fever virus isolates from the epizootic in The Netherlands in 1997-1998. AB - Sixteen classical swine fever virus (CSFV) field isolates from outbreaks of classical swine fever from the period between February 1997 and March 1998 in the Netherlands were sequence analysed. Parts of the 5' noncoding region (5'NCR) and the E1/E2 gene were sequenced after RT-PCR. The obtained sequences were compared with isolates of recent outbreaks in Europe and those of former outbreaks in the Netherlands. Sequence alignment of the 5'NCR region (321 bp) revealed that the isolates of the Dutch outbreak of 1997-1998 were closely linked to an isolate of the CSF outbreak that started in Paderborn, Germany in 1996. A relatively large fragment of the E1/E2 gene of 850 bp, including the antigenic region of E2, which is one of the most variable regions of the CSFV genome, was sequenced to determine whether this region can be used for epidemiology within an epizootic. Epidemiological tracing of transmission of virus was followed, starting from the first isolate and a line of five generations of viruses was analysed. Besides this, new isolates which could not be epidemiologically linked to preceding ones were also characterised. Differences between the isolates of the Dutch outbreak were minor both for the linked as well as for the non-linked isolates, indicating that all isolates have a common origin. Furthermore, our data show for the first time the genetic stability of CSFV even in the highly variable antigenic region of the E2 gene during a major epidemic lasting more than 1 year. PMID- 10384891 TI - Prevalence of calf diarrhea caused by bovine group A rotavirus carrying G serotype 8 specificity. AB - One hundred and seventeen rectal fecal specimens were collected in 1995 and 1996 from calves with diarrhea in Kagoshima Prefecture in Japan. The bovine group A rotavirus was detected by enzyme immunoassay in 43 of 117 specimens and isolated from 33 of the 43 specimens that were positive. G serotype, P serotype, and P genotype of 33 isolates were identified by reverse transcription-polymerase chain reaction, and 20 of 33 isolates (60.6%) were identified as G serotype 8. Thus, we discovered that calf diarrhea caused by bovine group A rotavirus carrying G serotype 8 specificity was prevalent in this research area during this research period. To our knowledge, this is the first report on the prevalence of calf diarrhea caused by the bovine group A rotavirus carrying G serotype 8 specificity. PMID- 10384892 TI - Epitopes and nuclear localization analyses of canine distemper virus nucleocapsid protein by expression of its deletion mutants. AB - A series of nucleocapsid protein (NP)-deleted genes of the Onderstepoort strain was constructed in order to locate antigenic regions of the NP of canine distemper virus. The expression of proteins from 5'-deleted NP genes was examined in COS-7 cells by indirect immunofluorescence assay using three monoclonal antibodies (MAbs), c-5, f-5 and h-6, and a rabbit serum against NP. These MAbs reacted with two regions of NP. Amino acid residues from 1 to 80, and 337-358, were necessary and sufficient for formation of the epitopes identified by MAbs f 5 and h-6, and c-5, respectively. The proteins translated from intact or 3' deleted genes were found to be localized in the nuclei of COS-7 cells, whereas the proteins from the 5'-deleted genes were mainly detected in the cytoplasm. These results suggested that 80 amino acid residues at the N-terminus are required for transportation of NP into the nucleus. PMID- 10384893 TI - Determination of acute noise effects using distortion product otoacoustic emissions. AB - Because distortion product otoacoustic emissions (DPOAE) are the product of the effect of two sinus tones on the cochlea, a multitude of combinations regarding the amplitude and the frequency ratio of the primary tones exists. The goal of the present study was to directly compare different stimulus combinations described in the literature in the detection of acute noise trauma using DPOAE. In the present study, 13 volunteers were exposed for 1 h to noise that was equivalent to sound levels in a discotheque. Audiograms and distortion product otoacoustic emissions were measured before and after noise exposure using four different stimulus combinations. For three of these settings, L1 was 65 dB, L1-L2 was 25 dB and f2/f1 was varied and set to 1.22, 1.20 and 1.18. For the fourth setting, L1 was at 65 dB, whereas L1-L2 was at 10 dB (f2/f1 = 1.20). A second group of volunteers (n = 14) was measured using identical time periods and setting, but was not exposed to noise. The comparison of different stimulus combinations showed that the stimulus combination L1 = 65 dB and L1-L2 = 25 dB at f2/f1 = 1.18 was best suited for detecting a difference between noise-exposed and unexposed individuals. PMID- 10384894 TI - Binaural interaction component in children at risk for central auditory processing disorders. AB - The binaural interaction component (BIC) occurring in the latency range of peak V of the auditory brainstem responses (ABR) was investigated in nine normal children, comprising the control group, and nine children at risk for central auditory processing disorders (CAPD), comprising the CAPD group. All children tested had normal hearing, normal intelligence and normal ABR thresholds. Averaged BIC obtained from the difference between the summed and binaural ABR waveforms was compared between the two groups for amplitude and latency measures. Results indicated a significant reduction in the amplitude of the BIC occurring in the latency domain of ABR peak V, in the CAPD group. PMID- 10384895 TI - Slope analysis of Auditory Brainstem Responses in children at risk of central auditory processing disorders. AB - The method of slope vectors was used to quantify Auditory Brainstem Responses (ABR) obtained from nine normal children and nine children at risk for central auditory processing disorders (CAPD) with language impairment, for monaural and binaural stimulation conditions. Slopes thus obtained were subjected to K-Means Cluster Analysis. Distinction between the two groups was obtained only for binaural stimulation conditions, wherein all normal children were grouped under cluster 1 with higher slope values and 6 out of 9 CAPD children were grouped under cluster 2 with lower slopes. The results suggest that there may be several subcategories among children who are found to be at risk for CAPD. One of the subcategories may comprise children who exhibit poor ABR morphology, especially during binaural stimulation conditions, which could be due to binaural interference. PMID- 10384896 TI - Long-term effect of hyperbaric oxygenation treatment on chronic distressing tinnitus. AB - Tinnitus is still a phenomenon with an unknown pathophysiology with few therapeutic measures. During the last two decades, hyperbaric oxygenation therapy (HBO) has been used in the treatment of sudden deafness and chronic distressing tinnitus. In this study, we prescribed HBO to 20 patients who had had severe tinnitus for more than one year and who had already had other forms of tinnitus therapy with unsatisfactory results. Four patients could not cope with the pressure gradient. The effect of HBO was assessed using subjective evaluation and VAS scores before and after HBO. Follow-up continued until one year after treatment. Six patients had a reduction of tinnitus and accompanying symptoms, eight patients did not notice any change and two patients experienced an adverse effect. Any outcome persisted with minor changes until one year after treatment. HBO may contribute to the treatment of severe tinnitus, but the negative effect on tinnitus should be weighed carefully. PMID- 10384897 TI - Distortion product oto-acoustic emissions as objective audiometry in a group of children with ventilation tubes. AB - The aim of this study was to evaluate distortion product oto-acoustic emissions (DPOAEs) as a means of objective audiometry in a population of children with ventilation tubes. We measured DPOAEs at two different stimulus levels--recorded transient evoked oto-acoustic emissions (TEOAEs) and obtained pure-tone audiometry (PTA). DPOAEs were compared with the normal range proposed by the Madsen company: the CELESTA 503 provides a 'normalized' distortion-product-gram which we compared with the pure-tone threshold of the test group. DPOAEs at 60 dBSPL were more easily obtained in the test group than TEOAEs and PTA. Correlation between 'normalized' DPOAEs and pure-tone thresholds was the strongest at 2 and 4 kHz. Surprisingly, DPOAEs at 60 dBSPL showed significant differences also at 2 and 4 kHz compared with the Madsen data. DPOAEs at 60 dBSPL might be an easy, objective test for evaluating auditory function and for determining hearing threshold at 2 and 4 kHz. PMID- 10384898 TI - Minimum Nordic requirements for clinical testing of hearing aids. Nordiska samarbetsorganet for handikappfragor (Nordic Co-operation on Disability). Working group for harmonization of requirements on aids for hearing-impaired persons, June 1998. PMID- 10384899 TI - Clinical study of Widex Senso on first-time hearing aid users. AB - Using psychoacoustic tests and questionnaires, the aim of this study was to clinically test Widex Senso (WS) versus analogue hearing aids on 200 first-time wearers. Half of the participants were selected at random for fitting with the behind-the-ear model (WS C8) or the in-the-canal model (WS CX). On a group basis, WS was found to provide more benefit than a palette of 29 analogue, modern hearing aid models from 10 manufacturers. Only 3 of 100 subjects changed from WS to another hearing aid. On average, the abbreviated profile of hearing aid benefit (APHAB) (Cox & Alexander, 1995) demonstrated superior performance for WS, i.e. no conflict existed between high comfort and high speech recognition. Median aided frequency-modulated tone thresholds in the sound field were better than 25 dB HL at frequencies up to 4 kHz inclusive. A distinct mean aided improvement of speech threshold in competing speech of 2.5 dB was found in both groups. PMID- 10384900 TI - Clinical study of a digital vs an analogue hearing aid. AB - Digital signal processing in hearing instruments has brought new perspectives to the compensation of hearing impairment and may result in alleviation of the adverse effects of hearing problems. This study compares a commercially available digital signal processing hearing aid (HA) (Senso) with a modern analogue HA with programmable fitting (Logo). The HAs tested are identical in appearance and, in spite of a different mode of operation, the study design ensured blinding of the test subjects. Outcome parameters were: improvements in speech recognition score in noise (deltaSRSN) with the HAs; overall preference for HA; overall satisfaction; and various measures of HA performance evaluated by a self assessment questionnaire. A total of 28 experienced HA users with sensorineural hearing impairment were included and 25 completed the trial. No significant differences were found in deltaSRSN between the two HAs. Eleven subjects indicated an overall preference for the digital HA, 10 preferred the analogue HA and 4 had no preference. Concerning overall satisfaction, 8 subjects rated the digital HA superior to the analogue one, whereas 7 indicated a superior rating for the analogue HA and 10 rated the HAs equal. Acceptability of noise from traffic was the only outcome parameter which gave a significant difference between the HAs in favour of the digital HA. It is concluded that there are no significant differences in outcome between the digital and analogue signal processing HAs tested by these experienced HA-users. PMID- 10384902 TI - Serological survey of antibodies to Toxoplasma gondii in goats, sheep, cattle and water buffaloes in Bahia State, Brazil. AB - Serum samples from 439 goats, 240 sheep, 194 cattle and 104 water buffaloes were tested for antibodies to Toxoplasma gondii by a latex agglutination test. Antibodies to T. gondii were found in 28.93% of goats, 18.75% of sheep, 1.03% of cattle and 3.85% of water buffaloes, at a dilution of 1:64. The highest titres observed in goats, sheep, cattle and water buffaloes were 1:2048, 1:2048, 1:64 and 1:512, respectively. PMID- 10384903 TI - Occurrence of Isospora suis in larger piglet production units and on specialized piglet rearing farms. AB - Mixed fecal samples of 264 litters from five piglet production farms (155-238 sows/farm) were investigated three times during the suckling period for the occurrence of Isospora suis over the period of 1 year. On all five farms Isopora suis was found to be a common endoparasite with infection rates being highest in litters of 3-4 weeks of age. By the end of the third investigation period the cumulative infection rate was 53.8% of the litters ranging from 20.0% to 81.5% for the single farms. During the suckling period the infection rate increased from 18.6% to 32.6% and then to 37.7%. Diarrhea was present in 66.3% of the sampled litters with the highest rates at the end of the suckling period. 63.4% of the litters which showed diarrhea and 34.8% of those without diarrhea excreted I. suis within the study period. Diarrhea was recorded for 78.2% of the I. suis positive litters and for 52.5% of the Isospora-negative litters. In summer and fall the occurrence of I. suis was higher (66.3% and 61.0%, respectively) than in spring and winter (47.7% and 37.9%, respectively). In litters with diarrhea and pathogenic E. coli I. suis often occurred simultaneously. Above-average hygiene measures and mainly perforated pen floors seemed to lower the risk of isosporosis. With the exception of Strongyloides ransomi other parasites were not found in the fecal samples of suckling piglets. Two specialized piglet rearing farms, a conventional large-scale rearing unit and a farm managed according to the segregated early weaning (SEW) system were examined three times during the 6 7 week rearing period. In both units I. suis was common, but was not correlated with diarrhea. In the SEW unit the infection rates decreased from 37.5% to 20.2% and to 4.1%, while the infection rate in the conventional unit slightly increased from the first (17.2%) to the second (21.9%) investigation and stayed at this level at the third sampling. PMID- 10384901 TI - Canine hepatozoonosis: comparison of lesions and parasites in skeletal muscle of dogs experimentally or naturally infected with Hepatozoon americanum. AB - We report previously undescribed, early lesions in skeletal muscle of dogs experimentally infected with Hepatozoon americanum by ingestion of laboratory reared, infected Amblyomma maculatum. The earliest muscle lesion was recognized at the first interval of examination 3 weeks following exposure. The lesion consisted of a large, modified host cell whose cytoplasm frequently contained a demonstrable parasite. In skeletal muscle, the cell was consistently located between muscle fibers or in loose connective tissue adjacent to those fibers. Evidence suggesting that the parasite arrives in muscle and other tissue within the host cell cytoplasm is presented. Mucopolysaccharide encystment of the host cell, absent at this early stage, was acquired gradually and approached maximal development 26 weeks post exposure. Completion of the asexual cycle as evidenced by the presence of parasites entering vascular lumens within granulomas and also by the presence of gamonts in peripheral blood leukocytes, occurred within 28-32 days postexposure. Progression of the parasite cycle from meront to passage of zoites into vessel lumens of granulomas can occur in 11 or fewer days. The density with which parasitic lesions occur in one named skeletal muscle compared to other named muscles, although somewhat variable, was not significantly different in either experimentally induced or natural infections. The distribution of developmental stages of the parasite/lesion in four experimental infections (969 lesions) is compared with those in eight dogs with natural infections (557 lesions). PMID- 10384904 TI - Primary experimental infection of riverine buffaloes with Fasciola gigantica. AB - The clinical course of the primary experimental Fasciola gigantica infection was investigated in riverine buffalo calves of the Murrah breed. Nine male calves aged 12-15 months were randomly assigned to two groups of five (Group I) and four (Group II) animals. Each animal in Group I, was orally infected with 1000 metacercariae (mc) of F. gigantica, whereas Group II animals did not receive any infection dose and served as uninfected controls. No clinical signs of fasciolosis were observed until the sixth week post-infection (PI). Group I animals, however, developed recognised symptoms of acute fasciolosis, comprising apyrexic inappetance, anemia, poor weight gain, diarrhoea and sub-mandibular and facial oedema, respectively, from 5, 6, 8, 16 and 17 weeks PI. The signs were intermittent in nature and of variable duration. The prepatent period was of 92 97 days (mean 95.2 +/- 3.1). One of the five infected animals died on Day 147 PI. At necropsy, 36.8 +/- 11.0% of the infection dose was recovered as adult fluke population. The gross lesions were primarily biliary in nature. Group II, the uninfected controls, throughout the study period of 165 days PI, did not show any symptom and were negative for F. gigantica. The study demonstrated that the onset of adverse effects of F. gigantica on the growth and health of the infected host was mainly noted during late prepatency much before coprological prediction and diagnosis. The significance of preventive therapy against fasciolosis during prepatency has been stressed in endemic areas. PMID- 10384905 TI - Persistent efficacy of doramectin pour-on against artificially induced infections of nematodes in cattle. AB - Two studies were conducted to determine the persistent efficacy of doramectin pour-on against an artificial, trickle challenge of mixed nematodes in calves. In each study, 42, 4-8 months old calves were randomly assigned into four groups of 10 animals each (T1-T4), plus two larval-viability monitor animals. All animals were treated with fenbendazole (10 mg kg(-1)) 14 days prior to the start of the study to clear any existing infection. Doramectin pour-on at 500 microg kg(-1) was used on each animal in Groups T2, T3, and T4 with intervals of 1 week (Day 0, 7, and 14, respectively). Calves in Group T1 were treated with saline solution on Day 0 and at the same volumetric rate (1 ml 10 kg(-1)) as the doramectin treated animals. All treatments were applied in a single passage along the midline of the back, from the withers to the tailhead. Subsequently, trickle inoculations with infective larvae were administered to all calves for 22 consecutive days (Days 14 35). Doramectin pour-on provided > or = 91.9% efficacy against challenge with Dictyocaulus viviparus, Haemonchus spp., and Ostertagia ostertagi for up to 35 days post-treatment and against challenge with Cooperia oncophora, Cooperia punctata, and Oesophagostomum radiatum for up to 28 days post-treatment. PMID- 10384907 TI - Efficacy of fenbendazole granules and pyrantel pamoate suspension against Toxocara canis in greyhounds housed in contaminated runs. AB - The efficacy of fenbendazole granules against Toxocara canis in naturally infected greyhounds housed in contaminated environments was evaluated. Eight pens, each containing three to seven greyhounds, 3-12 months of age, were randomly allotted into two treatment groups. Greyhounds in Group 1 were treated with fenbendazole granules mixed in their feed at 50 mg/kg/day for 3 consecutive days once a month for 4 months. Greyhounds in Group 2 were treated with pyrantel pamoate suspension at 5.0 mg/kg per os once a month for 4 months. Quantitative fecal examinations were performed on days 0, 10 and then on the first day of each monthly treatment. Greyhounds administered fenbendazole had fecal egg count reductions (FECRs) of 95.8 and 99.8% at 10 and 31 days following initial treatment, respectively. Greyhounds administered pyrantel pamoate had FECRs of 85.8 and 88.3% at 10 and 31 days after the first treatment, respectively. T. canis fecal egg counts conducted from Day 31 through Day 128 were significant lower in those greyhounds administered fenbendazole as compared to greyhounds administered pyrantel pamoate. Fenbendazole produced FECRs in greyhounds from Day 31 through Day 128 by 96.8-99.8%. Pyrantel pamoate reduced fecal egg counts during the same time period 71.4-98.3%. PMID- 10384906 TI - Anthelmintic resistance in sheep and goat farms on Peninsular Malaysia. AB - The faecal egg count reduction test (FECRT) was conducted on 39 sheep farms and 9 goat farms located in Peninsular Malaysia. The anthelmintic groups used in these tests were the benzimidazoles, levamisole, the benzimidazole/levamisole combination, macrocyclic lactones and closantel. Results indicated that the prevalence of resistance to the benzimidazole group was high, with approximately 50% of the sheep farms and 75% of the goat farms having resistant nematode parasite populations present. Resistance to levamisole, closantel and ivermectin was also detected. Differentiation of the infective larvae derived from faecal cultures indicated that by far the most predominant parasite species was Haemonchus contortus. PMID- 10384908 TI - Evaluation of a yearly insecticidal ear tag rotation for control of pyrethroid resistant horn flies (Diptera: Muscidae). AB - From 1991 to 1997, the yearly alternated use of synergized pyrethroid (lambda cyhalothrin + piperonyl butoxide) and organophosphate (pirimiphos-methyl) ear tags was evaluated for the control of two pyrethroid-resistant horn fly populations in Louisiana. At each site, weekly fly counts were used to assess product efficacy. Control achieved by synergized pyrethroid ear tag treatments was reduced from 7 to 2 weeks and from 4 to 0 weeks at St. Joseph and Winnsboro, respectively. Control by organophosphate ear tags decreased from 15 to 3 weeks and from 10 to 7 weeks at St. Joseph and Winnsboro, respectively. The rotation of synergized lambda-cyhalothrin and pirimiphos-methyl ear tags did not improve pyrethroid ear tag efficacy or prevent further development of resistance to the pyrethroid or OP compound. PMID- 10384909 TI - Confirmation that the dog is a definitive host for Neospora caninum. AB - Two mixed-breed littermate dogs were fed mouse brains containing tissue cysts of the NC-beef isolate of Neospora caninum. Both dogs excreted N. caninum oocysts in their feces. Dog 1 which was given methylprednisolone acetate (MPA) prior to ingesting tissue cysts, excreted oocysts on days 5 to 10 inclusive and on day 17 after ingesting tissue cysts. Dog 1 had a serum antibody titer of 1:200 in the indirect fluorescent antibody test (IFAT) 35 days after it was fed tissue cysts. Dog 2, which was not treated with MPA, excreted oocysts on Day 6 and Day 9 after ingesting tissue cysts. Antibodies to N. caninum were not found in a 1:25 dilution of serum on any examination period for Dog 2 during the study. Neospora caninum was not found in the tissues of either dog by histological or immunohistochemical means following necropsy 42 days after being fed tissue cysts. The identity of the oocysts excreted in the feces of the dogs was confirmed by mouse inoculation studies. PMID- 10384910 TI - What biochemical markers are best for measuring the extent of disease in patients with sclerosis? PMID- 10384911 TI - Inhibitory effect of vasoactive intestinal peptide on the challenge phase of allergic contact dermatitis in humans. AB - There is increasing evidence that the nervous system has influence on the immune response. The effect of vasoactive intestinal peptide (VIP) and of serotonin and its antagonists on the challenge phase of allergic contact dermatitis in humans were tested. The substances were injected intracutaneously shortly before and 6 h after application of patch tests with nickel sulphate in nickel-allergic patients and the test areas were measured after a further 18 h. Biopsy specimens were also taken for immunohistochemistry. The diameter of the nickel sulphate-induced test reaction was significantly reduced after injection of VIP at 10(-6)-10(-5) mol/l, but was not affected by serotonin or ketanserin. Also tested was the influence of the substances on the response of peripheral blood mononuclear cells from nickel allergic subjects to nickel sulphate, when added at the same time as the antigen. No effect on the cell proliferative rate was seen, except for an inhibitory effect of serotonin and its antagonists at 10(-5)-10(-4) mol/l. VIP, at 10(-5) mol/l and serotonin at 10(-4) mol/l stimulated the secretion of interferon gamma. The interleukin-2 soluble receptor secretion was slightly stimulated by 5-HT at 10(-4) mol/l and by ketanserin at 10(-6) mol/l. In conclusion, our results show that when injected intracutaneously in the challenge phase of allergic contact dermatitis, VIP has an inhibitory effect, which might be explained by enhanced leukocyte production of interferon gamma. PMID- 10384912 TI - Eccrine sweat glands: expression of transforming growth factor-beta and bone morphogenetic protein type I receptors and their intracellular signalling Smad proteins. AB - The transforming growth factor-beta superfamily is thought to be involved in the regulation and control of growth and differentiation. These growth factors signal through transmembrane serine/threonine kinase receptors. The activation of type I receptor kinase phosphorylates a family of intracellular signalling proteins called Smads. In the present study, we wanted to localize type I and type II receptors and Smad proteins in human eccrine sweat glands. Expression of transforming growth factor-beta type I receptor was restricted to myoepithelial cells only, whereas bone morphogenetic protein receptor IA was found selectively within the duct epithelium of both the dermal portion and the acrosyringium. Bone morphogenetic protein receptor IB antibody gave a faint staining of secretory epithelium and myoepithelial cells. Smad proteins were identified in different parts of the eccrine sweat gland apparatus. In particular, Smad 1 and Smad 3 were localized within myoepithelial cells, whereas coils were stained weakly for Smad 1 and Smad 3. Smad 3 protein was also expressed by the duct epithelium. Smad 2, Smad 4, Smad 5, Smad 6 and Smad 7 were not identified in eccrine sweat gland epithelia. Our data provide evidence for transforming growth factor-beta/bone morphogenetic protein signalling in the eccrine sweat gland and the selective expression of Smad proteins. Myoepithelial cells and duct cells have been identified as major targets of the transforming growth factor-beta pathway. Possible functions are growth inhibition and control of myoepithelial differentiation. PMID- 10384913 TI - Eicosanoid and cytokine levels in acute skin irritation in response to tape stripping and capsaicin. AB - We assessed the effects of physical and chemical irritants on a profile of acute inflammatory mediators in normal human skin. Skin damage in both cases is accompanied by a flux of inflammatory processes and repair mechanisms, which remain imprecisely understood. We used 10 sequential cellotape strips or topical application of 0.075% capsaicin as skin irritants and characterized the subsequent production and/or release of inflammatory mediators in suction blister fluids from human skin in vivo. In tape stripped skin, levels of prostaglandin E2 and interleukin-1alpha were increased 3.4-fold and 3.3-fold, respectively (p<0.0001; p<0.02), levels of tumour necrosis factor-alpha were decreased 3.0 fold (p<0.01), whereas levels of interleukin-6 and leukotriene B4 in blister fluids remained relatively unchanged. For the capsaicin-treated skin, levels of mediators showed only minor differences when compared with matched controls. However, a correlation was observed between levels of prostaglandin E2 and interleukin-1alpha in capsaicin pre-treated blister fluids (r=0.58, p<0.01, n=19). These data are consistent with prostaglandin E2 and interleukin-1alpha playing key roles in acute skin responses to mild irritants. PMID- 10384914 TI - Suction blister formation in skin after acute and repeated mast cell degranulation. AB - Mast cells and their proteases are thought to participate in the development of skin blisters in various pathological conditions. In this study, suction blistering was used as an experimental model to evaluate the significance of mast cells in blister formation after pre-treatment of normal skin with intradermal injections of 100 microg/ml compound 48/80 (a mast cell degranulator) or with 0.1% capsaicin cream. Tryptic and chymotryptic enzyme activities in blister fluids were measured with sensitive p-nitroanilide substrates. Repeated injections of compound 48/80 once a day on 3 or 5 consecutive days or capsaicin applications 3 times a day for 7 or 10 days were used to induce mast cell degranulation and inflammation in normal skin. Both treatments ultimately led to decreased wheal and erythema reactions before suction blistering, but neither treatment affected the size or formation rate of suction blisters. No suction blister fluids had detectable levels of chymotryptic activity, but blister fluids from bullous pemphigoid, herpes zoster and insect bullous eruption, used as the control, revealed clear chymotryptic activity. In addition, tryptic activity in suction blister fluids was not significantly altered after compound 48/80 and capsaicin pre-treatments. However, if the wheal reaction was induced immediately before suction blistering, a significantly increased rate in blister formation together with increased tryptic activity was found, but, unexpectedly, no chymotryptic activity could be detected in blister fluids. The results show that repeated mast cell degranulation in normal skin has no effect on the formation rate of suction blisters, which developed more rapidly on acutely whealing skin. This is probably due to skin oedema rather than mast cell proteases, since no chymotryptic activity was detected in suction blisters where tryptic activity exhibited high individual variation. PMID- 10384915 TI - Expression of p53 protein before and after PUVA treatment in psoriasis. AB - We investigated the effect of the potentially carcinogenic psoralen plus UVA radiation (PUVA) therapy on the expression of p53 in skin of psoriatic patients. p53 antibodies DO7 and Pab240, antibodies against PCNA and Ki67 and the avidin biotin immunoperoxidase complex method were used in the immunohistochemical staining of biopsy samples from non-lesional and lesional skin of 23 patients who received either trioxsalen bath PUVA or oral 8-methoxypsoralen PUVA. Biopsies were taken before and after a PUVA course. A modest expression of p53 was seen in psoriatic lesions in 17/21 patients before any treatment, probably as a physiological reaction to the hyperproliferation. Both p53 and the proliferation markers Ki67 and PCNA followed the same pattern, being more frequent in psoriatic lesions than in non-lesional skin. Exposure to PUVA induced an increase in p53 expression in non-lesional skin in 14/19 patients, putatively as a response to DNA damage caused by PUVA. In psoriatic lesions about half of the patients showed increased and half decreased expression of p53. The latter finding might be explained by decreased proliferation activity of the healing epidermis. In conclusion, p53 nuclear positivity in non-lesional skin after PUVA treatment is likely to be induced by DNA damage caused by PUVA, while in psoriatic lesions it could be a result of the combined effect of decreasing epidermal proliferation and DNA-damage. PMID- 10384916 TI - Epidermal urocanic acid concentration and photoisomerization reactivity in patients with cutaneous malignant melanoma or basal cell carcinoma. AB - The relationship of epidermal urocanic acid concentration and photoisomerization reactivity to human skin cancer was studied. Twelve cutaneous malignant melanoma patients, 10 basal cell carcinoma patients and 22 healthy matched controls were enrolled in the study. A solar simulating ultraviolet irradiator was used for phototesting the minimal erythema dose. Using the Finn Chamber technique, urocanic acid was sampled from the healthy skin of the upper back, prior to and after exposure to suberythemal UV doses. The mean values of total and trans urocanic acid were higher in basal cell carcinoma patients than in controls, but this difference was not statistically significant. No corresponding phenomenon was evident in the case of cutaneous malignant melanoma patients and their controls. Photoisomerization induced by irradiation with 1 mJ/cm2 CIE (Commission Internationale de l'Eclairage) was statistically significantly lower in cutaneous malignant melanoma patients than in controls (p=0.04). A similar trend was seen in basal cell carcinoma patients vs. their controls, but the difference was not significant. PMID- 10384917 TI - Photodynamic therapy by topical aminolevulinic acid, dimethylsulphoxide and curettage in nodular basal cell carcinoma: a one-year follow-up study. AB - Fifty-eight patients with 119 nodular (2 mm or more in thickness) basal cell carcinomas successfully treated with photodynamic therapy were included in this 1 year follow-up study. The initial cure rate at 3-6 months was 92% after photodynamic therapy, which included an initial debulking procedure and topical application of dimethylsulphoxide in order to enhance penetration of 5 aminolevulinic acid (20% in cream) to which the lesions were exposed for 3 h prior to exposure to light. At examination 12-26 months (mean 17 months) after treatment 113 lesions (95%) were still in complete response. Six lesions (5%) had recurred, located on the face, scalp and ear. The cosmetic outcome was evaluated as excellent to good in 91%. Microscopic examination of biopsies taken from healed areas in 7 patients did not reveal any sign of damage in 5 and only minor alterations in 2. PMID- 10384918 TI - Serum levels of soluble TNF alpha receptor type I and the severity of systemic sclerosis. AB - Several abnormalities of cytokines have been shown to occur in systemic scleroderma; however their correlation with clinical parameters is controversial. Since serum concentrations of cytokine receptors have been shown to correlate with inflammatory processes, including systemic sclerosis, the aim of our study was to compare serum concentrations of TNF alpha receptor type 1 with the concentrations of soluble intercellular adhesion molecule-1 (sICAM-1), soluble interleukin-2 receptor (sIL-2R) and aminoterminal propeptide of procollagen type III (PIII NP). The findings were correlated with the clinical parameters and antibody patterns, and with the disease severity. Serum samples were studied with the use of enzyme-linked immunosorbent assay (ELISA) or radioimmunoassay (RIA) for sTNF alpha R1, sICAM-1, sIL-2R and PIII NP. The series comprised 36 patients with systemic scleroderma: 13 with diffuse variety and 23 with limited variety, and 7 with Raynaud's disease. Healthy volunteers (n = 25) were chosen from doctors and/or other laboratory staff. Increased levels of sTNF alpha-receptor type 1 were found in 77% of patients with diffuse variety and in only 30% of patients with limited form. Increased serum concentrations in patients with diffuse scleroderma and limited variety were found for sICAM-1 54% and 65%, for sIL-2R 46% and 15%, and for PIIINP 77% and 50%, respectively. There were significant correlations between serum levels of sTNF alpha-receptor type 1 and PIIINP (r = 0.653, p < 0.0001), and sTNF alpha-receptor type 1 and sIL-2R (r = 0.625, p < 0.0001), but not between sTNF alpha-receptor type 1 and sICAM-1 (r = 0.127, p < 0.526). Clinical analysis revealed that serum concentrations of sTNF alpha-RI seem to correlate best with the severity of the disease and, as the only parameter, correlated with lung involvement. The study showed that, in addition to recognized parameters of scleroderma severity (IL-2R, PIIINP), a new important marker appears to be sTNF alpha-receptor type 1. PMID- 10384919 TI - Sunscreen sensitization: a 5-year study. AB - The purpose of this study was to evaluate the prevalence of sunscreen contact allergy and/or contact photoallergy in 370 patients with suspected photodermatitis. Patch and photopatch tests were performed using the French Society of Photodermatology (SFPD) standard series. A total of 57 cases of contact allergy and/or photocontact allergy to sunscreens were diagnosed (15.4%). Amongst these, 27 reactions were related to oxybenzone and 14 to isopropyl dibenzoylmethane. These results, obtained from January 1990 to December 1994, confirm that, given the high frequency of photosensitization cases, a large part of the battery of photopatch tests should be dedicated to sunblocks. PMID- 10384920 TI - Quantitative image analysis of hair follicles in alopecia areata. AB - We took biopsies from similar sites on the scalps of normal controls, normal looking and hair loss areas of patients with alopecia areata. The specimens were sectioned serially and horizontally. We measured morphological parameters, such as the diameter of the hair shaft, the thickness of the inner root sheath, the diameter of the outer root sheath, the irregularity of the outer root sheath and the ratio between the diameters of the hair shaft and the outer root sheath, using a computerized image analysis system. There were significant differences in 5 parameters between hair loss areas and normal controls. The proportions of vellus and telogen hair were significantly higher in the areata areas than in the controls. The 5 morphological parameters and most quantitative-measured parameters of normal-looking areas from patients with alopecia values intermediate between the other groups. PMID- 10384921 TI - Cultured allogeneic skin cells are effective in the treatment of chronic diabetic leg and foot ulcers. AB - Diabetic ulcers on the lower extremities present a difficult treatment problem, and some ulcers respond poorly to conventional topical and cast treatment. The purpose of this study was to assess the effect of cultured allogeneic keratinocyte epithelium and fibroblast-gelatin sponge on the healing of chronic, refractory diabetic leg and foot ulcers. Non-diabetic chronic leg ulcers were treated for comparison. This open study comprised 22 patients with type I or type II diabetes and 16 patients with leg or ankle ulcers of different aetiologies. A total of 26 diabetic and 25 non-diabetic ulcers were treated mainly with keratinocyte epithelium and/or fibroblast-gelatin sponge once weekly until complete healing or until no further healing could be observed despite several repeated treatments. The duration of diabetic ulcers was 10.3+/-15.8 (mean+/-SD) months and the size 3.1+/-6.6 cm2. The diabetic ulcers were located in the heel (7), toe (7), sole (5), leg (6) and Achilles (1). The mean duration of non diabetic ulcers was 6.8+/-6.0 months and the size 10.5+/-11.8 cm2. A total of 12+/-11 skin cell transplantations were performed for the diabetic ulcers. All but 1 diabetic ulcer healed during the study. The time for 50% reduction in ulcer area was 32+/-32 days, but 99+/-110 days were needed for complete ulcer closure. The longer the ulcer had existed the longer was the healing time. Heel ulcers showed significantly slower healing response than leg, sole and toe ulcers. Preliminary results suggest that both keratinocytes and fibroblasts are equally effective in the healing process. The time required for healing of the diabetic ulcers did not differ markedly from that of the non-diabetic ulcers. The results suggest that cultured allogeneic skin cells used once weekly are effective in the treatment of recalcitrant diabetic ulcers. PMID- 10384922 TI - Randomized double-blind comparison of short-term itraconazole and terbinafine therapy for toenail onychomycosis. AB - Previous studies evaluating short-term itraconazole and terbinafine therapy for onychomycosis have varied in protocol and size; this double-blind study enabled a large-scale, standardized, direct comparison. Patients with toenail onychomycosis were randomized to itraconazole 200 mg daily (n = 146) or terbinafine 250 mg daily (n = 146) for 12 weeks, with a 36-week follow-up. Mycological cure rates at the follow-up end-point were significantly equivalent (61% with itraconazole vs. 67% with terbinafine). A similar proportion of patients in each group experienced adverse events during treatment (itraconazole, 22%; terbinafine, 23%). More patients receiving terbinafine stopped treatment permanently because of treatment related adverse events (8% vs. 1%). PMID- 10384923 TI - Treatment of digital mucous cysts with a carbon dioxide laser. AB - Digital mucous cysts are common tumours of the distal interphalangeal joint, causing pain, cosmetic disfigurement and nail deformities. This study describes six patients suffering from digital mucous cysts, some of which recurred after surgery. Carbon dioxide laser vaporization was performed under local anaesthesia, resulting in complete remission in four out of six patients. In two patients, the mucous cyst recurred within 3 weeks and 11 months, respectively, after laser therapy. Although complete remission cannot be achieved in all patients, carbon dioxide laser vaporization of mucous cysts is fast and easy to perform. No side effects of therapy, such as nail deformities, pain or infection, occurred. For this reason, more aggressive treatments, such as radical excision of the cyst, could be restricted to cases in which carbon dioxide laser therapy fails. PMID- 10384924 TI - HPV prevalence in anal warts tested with the MY09/MY11 SHARP Signal system. AB - Anal warts are, from an aetiological point of view, a diverse category of lesions including condylomata acuminata, fibroepithelial polyps and seborrhoeic keratosis. Human papillomavirus induced anal warts, in contrast to other types of warts, are contagious and not infrequently sexually transmitted, they therefore need to be accurately identified. A total of 24 anal warts were randomly collected and the histopathological diagnoses based on microscopy, alone or in combination with a sensitive PCR-based human papillomavirus test, were compared using the SHARP Signal system for detection. Three lesions were identified as condyloma acuminatum by morphology alone due to the obvious presence of koiloytotic atypia; 11 warts without koilocytes were identified only after a positive test for anogenital human papillomavirus. One additional lesion contained human papillomavirus DNA of cutaneous type and 9 papillomas were human papillomavirus-negative and tentatively diagnosed as fibroepithelial polyps or seborrhoeic keratosis. All 14 condylomas contained human papillomavirus of low risk type. Of these, 12 warts showed a positive human papillomavirus reaction with in situ hybridization. Morphology alone cannot reveal the true nature of most anal papillomas, even when koilocytotic atypia is considered as a diagnostic hallmark. An optimal diagnosis of anal warts requires a sensitive PCR-based human papillomavirus DNA test. A test for identification of cutaneous human papillomavirus DNA is also worthwhile. PMID- 10384926 TI - Drug-induced erythema multiforme-like bullous pemphigoid. PMID- 10384925 TI - Irritancy of scrubbing up for surgery with or without a brush. AB - Hand washing is an indispensable procedure for surgical nurses. Although scrubbing up with a brush is preferable to prevent infections, it is not clear how irritating to the skin scrubbing with a brush is compared with hand washing without a brush. TEWL, high frequency conductance and pH were measured on the hand skin of the same group of nurses before and after daily hand washing for 11 days in different seasons, which were chosen as favourable and unfavourable periods for the condition of hand skin, namely the early summer and autumn. Additionally, we compared the antimicrobial effects on the skin of scrubbing up, using a palm stamp method. TEWL showed significantly higher values with brush washing than with simple hand washing only in the autumn. There was no significant difference in the measurement of high frequency conductance, pH or in the antimicrobial effects between the two washing techniques. Results showed the deleterious effects on the skin of hand washing, particularly that of using a brush in the cold season. PMID- 10384927 TI - Cardiovascular complications of psoriasis: does obstructive sleep apnoea play a role? PMID- 10384928 TI - Paternal and maternal atopic dermatitis have the same influence on development of the disease in children. PMID- 10384929 TI - Cicatricial alopecia following therapeutic embolization. PMID- 10384931 TI - Angiodermatitis associated with congenital arteriovenous malformation on the elbow. PMID- 10384930 TI - Syringocystadenoma papilliferum without an antecedent naevus sebaceous. PMID- 10384932 TI - Pityriasis lichenoides chronica with acral distribution mimicking palmoplantar syphilid. PMID- 10384933 TI - White, fibrous, papular lesions associated with systemic lupus erythematosus: is this an ongoing scar following vascular involvement? PMID- 10384934 TI - Spiegler-Fendt type lymphocytoma cutis: a case report of two patients successfully treated with interferon alpha-2b. PMID- 10384935 TI - Lichen simplex chronicus with a cutaneous horn. PMID- 10384936 TI - Cutaneous alternariosis due to Alternaria chlamydospora after bone marrow transplantation. PMID- 10384937 TI - Secondary anetoderma in people with Down's syndrome. PMID- 10384938 TI - Difference in ceramide composition between "dry" and "normal" skin in patients with atopic dermatitis. PMID- 10384939 TI - Clinical and virological comparison of 3 patients with erythema multiforme. PMID- 10384940 TI - Mycobacterium avium complex: cutaneous infection in an immunocompetent host. PMID- 10384941 TI - Herpetic pharyngitis with mammary and genital herpes due to sexual contact. PMID- 10384942 TI - Angiosarcoma of the lower leg in chronic lymphoedema. PMID- 10384943 TI - Fournier's gangrene: a case report. PMID- 10384944 TI - Buschke-Loewenstein tumour is not a low-grade carcinoma but a giant verruca. PMID- 10384945 TI - The influence of diet and other factors on owner-perceived obesity in privately owned cats from metropolitan Perth, Western Australia. AB - A randomly selected group of cat-owning households (n = 458) were interviewed to determine the diet of their cats (n = 644) in the week prior to the survey and to identify dietary and other factors which were associated with obesity. All cats were categorised by their owners as underweight, correct-weight or overweight and the weight of 127 cats was also recorded. Nearly all cats were fed commercially prepared dry pet food (90.5%) or canned pet food (84.6%) in the week prior to the survey. Nineteen percent of cats were classified as overweight. Although the make up of a cat's diet was found not to be associated with its weight or weight category, cats fed dietary supplements or those which had not received a specific kitten diet when <6 months of age were more likely to be overweight after univariable analysis. Logistic multiple regression was used to investigate the effect of putative risk factors on obesity while controlling for other factors. Overweight cats were more likely to be cross-bred (OR = 2.1), neutered (OR = 2.8), living in houses with only one or two cats (OR = 1.8), male (OR = 1.4) and predominantly confined inside a house (OR = 1.4). Obesity is influenced by a variety of factors including host, dietary and management factors and these must be considered when developing weight control programmes for cats. PMID- 10384947 TI - The role of the public sector in the development and implementation of animal health policies. AB - Although it is widely accepted that both the public and private sectors have a role to play in improving animal health, the debate centers on the balance between the two. The comparative advantages of each sector depend on: (i) whether the targeted disease can affect humans; (ii) its degree of contagiousness; (iii) whether it is endemic or exotic; and (iv) the economic costs associated with the disease. The rationale for public intervention is weaker for non-contagious than for contagious diseases; governments, though, can play a support role in several areas: e.g. generation and dissemination of information on health management, fostering participation of producers' organization in the eradication of endemic diseases, or helping private research institutions and funders to overcome the hurdles posed by widespread uncertainty and high costs associated with basic research. Control and eradication of contagious diseases in characterized by strong externalities; bio-security measures implemented by any producer affect his/her neighbors. A major factor affecting the design of appropriate health policies for contagious diseases is whether the disease is endemic or exotic in a particular population. The externality exists for endemic diseases--but for exotic diseases there is only the possibility of an externality (which materializes solely in the case of an outbreak). For exotic diseases, therefore, the perception of the risk of an outbreak is a major determinant of producers' behavior and of public prevention policies. The perception by producers and policy makers of the probability of occurrence of an outbreak of an exotic disease depends on the time elapsed since the last outbreak in the country or in neighboring countries. In general, perception of the risk of an outbreak will be lower than the true risk for most exotic diseases that have been absent for many years--but might be higher than the true risk if an outbreak was recently reported in the region. Taxes and private insurance have been proposed to internalize the externality; however, these policies cannot solve the health externality. Alternative programs (such as joint public-private eradication campaigns) are proposed as a means to minimize the externality. PMID- 10384946 TI - Health and growth of water-buffalo calves in Nueva Ecija, the Philippines. AB - This prospective observational study was undertaken in the province of Nueva Ecija in the Philippines to assess current levels of health and growth achieved by a defined cohort of water-buffalo calves raised by smallholder farmers and to identify factors associated with the performance of these animals during the first 6 months following birth. Seventy two animals were enrolled, including 16 Philippine native water-buffalo and 54 crossbred (F1, F2 or backcrosses) animals. Dullness (which was associated with manual assistance at birth and inadequate milk supply subsequently) and scouring were the main signs of morbidity in this cohort, and the crude morbidity and mortality rates during the first 6 months following birth were 2.9 cases and 0.31 deaths per 1000 calf-days at risk, respectively. Average daily gain was significantly influenced both by the breed of the calf and whether the calf developed dullness at any time during the period of observation. The results of this study suggest that the problem of dullness in water-buffalo calves deserves further research attention. Management procedures are likely to be important determinants of growth in these animals. PMID- 10384948 TI - Prevalence of ketonemia and associations with herd size, lactation stage, parity, and postparturient diseases in Jordanian dairy cattle. AB - The prevalence of bovine ketonemia among 1155 dairy cows in various stages of lactation and parity on 25 Jordanian dairy herds was studied. The cross-sectional study was conducted during the spring of 1992. Serum concentration of beta hydroxybutyrate (BHB) <0.9 mmol/l, between 0.9 and 1.7 mmol/l, and >1.7 mmol/l were considered to indicate normal, mild and severe ketonemia, respectively. The point prevalences of mild and severe ketonemia were 22% and 3.8%, respectively. The prevalence of ketonemia decreased with increasing herd size. Associations between the prevalence of ketonemia and parity, stage of lactation, metritis, somatic-cell count (SCC) and serum cholesterol levels were not significant (p > 0.05). PMID- 10384950 TI - Modelling radiation-induced biological lesions: from initial energy depositions to chromosome aberrations. AB - The development of biophysical models of chromosome aberration induction has undergone considerable improvements in the past few years. This is mainly due to the development of new experimental techniques, such as fluorescence in situ hybridization (FISH) and premature chromosome condensation (PCC), and to a better knowledge of track structure characteristics (both in the physical and chemical stages). In particular, track structure simulations, providing a detailed description of the spatial distribution of energy depositions and relevant DNA lesions, represent a useful starting point for the development of 'ab initio' models. Various aspects of the processes determining the induction and the formation kinetics of chromosome aberrations are still under debate, concerning in particular the target description (interphase chromosome organization), the characterization of relevant DNA lesions, the possibility of inducing exchanges starting from single radiation-induced lesions, the rejoining mechanisms (proximity effects and possible induction of incomplete exchanges, i.e. one-way exchanges) and the influence of specific scoring criteria adopted both in experiments and models. Starting from Lea's breakage-and-reunion theory and Revell's exchange theory, an overview is given of various models recently reported in the literature. The assumptions adopted by the authors concerning the various processes involved in aberration formation are analysed in detail, in order to clarify the different approaches adopted in treating the open questions outlined above. PMID- 10384949 TI - Factors influencing recovery from and duration of lameness in Michigan (USA) horses. AB - The objective of this study was to identify factors that may affect recovery from and duration of a case of lameness in a stratified random sample of Michigan horses. This was done using data from Phase-II of the Michigan equine monitoring system (MEMS Phase-II), the equine health-monitoring study [Kaneene et al., Prev. Vet. Med. 29 (1997b) 277-292; Ross and Kaneene, Prev. Vet. Med. 28 (1996a) 209 224; Ross and Kaneene, Prev. Vet. Med. 29 (1996b) 59-75; Ross et al., Am. J. Vet. Res. 59 (1997) 23-29]. In this study, statistical modelling was conducted to evaluate risk factors affecting recovery from and duration of lameness using multivariable logistic regression and Cox's proportional hazards regression, respectively. Of 357 incident lameness cases reported during MEMS Phase-II, 280 (78.6%) were reported to have recovered. The median duration of a lameness case was 18 days (1st quartile (Q): 1, maximum (Max): 360). A total of 296 of 357 (82.9%) incident lameness cases received some type of treatment. Of 619 total treatments used, 329 (53.2%) were administered, conducted or applied by a veterinarian. Horses experiencing other types of lameness were less likely to recover than those experiencing hoof lameness (odds ratio (OR) = 0.48; 95% CI: 0.25, 0.93). Horses that had participated in exercise-related activities during the study period and prior to the lameness were more likely to recover (OR = 1.91; 95% CI: 1.05, 3.50). Treatment of the lameness was associated with an increased likelihood of recovery (OR = 1.82; 95% CI: 0.97, 3.45). Cases with a veterinarian involved in the diagnosis were associated with a decreased risk of recovery (OR = 0.48; 95% CI: 0.27, 0.84) and a longer duration lameness (HR = 0.58; 95% CI: 0.45, 0.73)--which might indicate that these cases were more complex or severe. Although cases treated for lameness were more likely to recover (OR = 1.82; 95% CI: 1.05, 3.50), treatment was not associated with lameness duration (HR = 0.58; 95% CI: 0.45, 0.73). PMID- 10384951 TI - Practical aspects of Monte Carlo simulation of charged particle transport: mixed algorithms and variance reduction techniques. AB - This paper is concerned with the practical implementation of Monte Carlo simulation methods for charged particle transport. The emphasis is on light particles (electrons and positrons) because of the larger scattering and energy straggling effects. Differential cross sections (DCS) for the various interaction mechanisms are described. As the average number of interactions along the particle track increases with the initial energy, detailed simulation becomes unfeasible at high energies. We can then rely on mixed simulation algorithms: hard events (i.e. individual interactions with angular deflection or energy loss larger than given cutoff values) are sampled from the DCS whereas soft events are simulated by means of a multiple scattering approach. Too frequently, the statistical uncertainty of analogue simulation (i.e. strict simulation of the physical interaction process) is found to be so large that results are meaningless. This problem can be partially solved by applying simple variance reduction techniques. PMID- 10384952 TI - DNA conformation and dynamics. AB - Nucleotide conformation and dynamics are important for the study of radiation damage to DNA at the atomic level. It is necessary to study not only normal oligonucleotide structure but also those containing modified bases which result from interaction with OH-radicals. There are now over 8000 atomic coordinate entries in the Brookhaven Protein Data Bank, of which over 900 relate to experimentally determined structures of nucleic acids and nucleic acid/protein complexes. We review some of these data which have led to the elucidation of novel DNA conformations, insight into DNA sequence specificity and knowledge of protein/DNA interactions. Further understanding of the conformation, stability and dynamics of nucleic acids has come from molecular modelling. We have used such techniques to study chemical modifications to bases such as alkylation of thymine and guanine and the effects of curvature in longer sequences. Recent improvements in this area include the inclusions of explicit counter-ions and solvent molecules, the use of Particle Mesh Ewald methods to incorporate the long range electrostatic interactions and the use of longer time scale simulations. We have employed these methods to analyse the effects of incorporation of 8 oxodeoxyguanosine into duplex DNA. This lesion is a common result of radiation damage and is known to have important effects in mutagenesis, cancer and ageing. PMID- 10384953 TI - Quantitative modelling of DNA damage using Monte Carlo track structure method. AB - This paper presents data on modelling of DNA damage induced by electrons, protons and alpha-particles to provide an insight into factors which determine the biological effectiveness of radiations of high and low linear energy transfer (LET). These data include the yield of single- and double-strand breaks (ssb, dsb) and base damage in a cellular environment. We obtain a ratio of 4-15 for ssb:dsb for solid and cellular DNA and a preliminary ratio of about 2 for base damage to strand breakage. Data are also given on specific characteristics of damage at the DNA level in the form of clustered damage of varying complexity, that challenge the repair processes and if not processed adequately could lead to the observed biological effects. It is shown that nearly 30% of dsb are of complex form for low-LET radiation, solely by virtue of additional breaks, rising to about 70% for high-LET radiation. Inclusion of base damage increases the complex proportion to about 60% and 90% for low- and high-LET radiation, respectively. The data show a twofold increase in frequencies of complex dsb from low-LET radiation when base damage is taken into account. It is shown that most ssb induced by high-LET radiation have associated base damages, and also a substantial proportion is induced by low-energy electrons. PMID- 10384954 TI - Simulation of DNA fragment distributions after irradiation with photons. AB - The Monte Carlo track structure code PARTRAC has been further improved by implementing electron scattering cross-sections for liquid water and by explicitly modelling the interaction of water radicals with DNA. The model of the genome inside a human cell nucleus in its interphase is based on the atomic coordinates of the DNA double helix with an additional volume for the water shell. The DNA helix is wound around histone complexes, and these nucleosomes are folded into chromatin fibres and further to fibre loops, which are interconnected to build chromosomes with a territorial organisation. Simulations have been performed for the irradiation of human fibroblast cells with carbon K and aluminium K ultrasoft x-rays, 220 kVp x-rays and 60Co gamma-rays. The ratio single-strand breaks to double-strand breaks (ssb/dsb) for both types of ultrasoft x-rays is lower than for gamma-rays by a factor of 2. The contributions of direct and indirect effects to strand break induction are almost independent of photon energy. Strand break patterns from indirect effects reflect differences in the susceptibility of the DNA helix to OH* attack inside the chromatin fibre. Distributions of small DNA fragments (<3 kbp) are determined by the chromatin fibre structure irrespective of whether direct or indirect effects are causing the breaks. In the calculated fragment size distributions for larger DNA fragments (>30 kbp), a substantial deviation from random breakage is found only for carbon K irradiation, and is attributed to its inhomogeneous dose distribution inside the cell nucleus. For the other radiation qualities, the results for larger fragments can be approximated by random breakage distributions calculated for a yield of dsb which is about 10% lower than the average for the whole genome. The excess of DNA fragments detected experimentally in the 8-300 kbp region after x-ray irradiation is not seen in our simulation results. PMID- 10384955 TI - Synthetic activity of rat blood lymphocytes under acute and continuous gamma irradiation -- fluorescent microspectral study. AB - The effects of different doses of acute and continuous gamma-irradiation on the synthetic activity of rat blood lymphocytes stained with acridine orange were studied by fluorescent microspectrometry. Male rats were exposed to acute gamma irradiation with doses of 7.5, 4 and 3 Gy, or to continuous irradiation with dose rates of 14.4, 2.1, 1.1 and 0.43 cGy/day, respectively. The changes of the synthetic activity of blood lymphocytes occurred in three main stages after acute gamma-irradiation and in four stages under continuous irradiation. The stages reflect the processes of depression and activation of the immune system under irradiation. Essential differences between the acute and continuous effects were observed in the first stage. After acute gamma-irradiation, the synthetic activity decreased sharply, indicating the predominant contribution of the damaging effect of irradiation, whereas under continuous irradiation, as a result of the stimulatory effect of low-dose irradiation, the synthetic activity increased during the first stage. PMID- 10384956 TI - Radon-induced lung cancer in smokers and non-smokers: risk implications using a two-mutation carcinogenesis model. AB - Three sets of data (population statistics in non-smokers, data from an investigation of the smoking habits of British doctors and a study of Colorado uranium miners) were used to analyse lung cancer in humans as a function of exposure to radon and smoking. One of the aims was to derive implications for radon risk estimates. The data were analysed using a two-mutation radiation carcinogenesis model and a stepwise determination of the model parameters. The basic model parameters for lung cancer were derived from the age dependence fit of the spontaneous lung cancer incidence in non-smokers. The effect of smoking was described by two additional parameters and, subsequently, the effect of radon by three other parameters; these five parameters define the dependence of the two mutation steps on smoking and exposure to radon. Using this approach, a consistent fit and comprehensive description of the three sets of data have been achieved, and the parameters could, at least partly, be related to cellular radiobiological data. The model results explain the different effect of radon on non-smokers and smokers as seen in epidemiological data. Although the analysis was only applied to a limited number of populations, lung cancer incidence as a result of radon exposure is estimated to be about ten times higher for people exposed at the age of about 15 than at about 50, although this effect is masked (especially for smokers) by the high lung cancer incidence from smoking. Using the model to calculate the lung cancer risks from lifetime exposure to radon, as is the case for indoor radon, higher risks were estimated than previously derived from epidemiological studies of the miners' data. The excess absolute risk per unit exposure of radon is about 1.7 times higher for smokers of 30 cigarettes per day than for non-smokers, even though, as a result of the low spontaneous tumour incidence in the non-smokers, the excess relative risk per unit exposure for the smokers is about 20 times lower than for the non-smokers. This prediction could have serious consequences for the transfer of risk estimates between populations. Although the solution of the model presented here is not unique but dependent on the model assumptions, the predictions and risk implications are sufficiently supported to justify a thorough investigation of the applicability of the model to other radon data sets. PMID- 10384957 TI - Biosynthesis of glycosylphosphatidylinositols in mammals and unicellular microbes. AB - Membrane anchoring of cell surface proteins via glycosylphosphatidylinositol (GPI) occurs in all eukaryotic organisms. In addition, GPI-related glycophospholipids are important constituents of the glycan coat of certain protozoa. Defects in GPI biosynthesis can retard, if not abolish growth of these organisms. In humans, a defect in GPI biosynthesis can cause paroxysmal nocturnal hemoglobinuria (PNH), a severe acquired bone marrow disorder. Here, we review advances in the characterization of GPI biosynthesis in parasitic protozoa, yeast and mammalian cells. The GPI core structure as well as the major steps in its biosynthesis are conserved throughout evolution. However, there are significant biosynthetic differences between mammals and microbes. First indications are that these differences could be exploited as targets in the design of novel pharmacotherapeutics that selectively inhibit GPI biosynthesis in unicellular microbes. PMID- 10384958 TI - Activation of DNA replication in yeast by recruitment of the RNA polymerase II transcription complex. AB - Activators of transcription are known to also play an important and direct role in activating DNA replication. However, the mechanism whereby they stimulate replication has remained elusive. One model suggests that, in the context of replication origins, transcriptional activators work by interacting with replication factors. We show that a defined, single interaction between a DNA bound derivative of the activator Gal4 and Gal11P, a mutant form of the RNA polymerase II holoenzyme component Gal11, suffices for stimulating DNA replication as it does for transcription. Moreover, recruitment of TBP, which can activate transcription from a gene promoter, also stimulates DNA replication from an origin site. These results strongly argue that transcriptional activators may not necessarily need to contact DNA replication factors directly, but can stimulate replication by recruiting the RNA polymerase II transcription complex to DNA. PMID- 10384959 TI - On the role of symmetrical and asymmetrical chaperonin complexes in assisted protein folding. AB - The cylindrical chaperonin GroEL of E. coli and its ring-shaped cofactor GroES cooperate in mediating the ATP-dependent folding of a wide range of polypeptides in vivo and in vitro. By binding to the ends of the GroEL cylinder, GroES displaces GroEL-bound polypeptide into an enclosed folding cage, thereby preventing protein aggregation during folding. The dynamic interaction of GroEL and GroES is regulated by the GroEL ATPase and involves the formation of asymmetrical GroEL:GroES1 and symmetrical GroEL: GroES2 complexes. The proposed role of the symmetrical complex as a catalytic intermediate of the chaperonin mechanism has been controversial. It has also been suggested that the formation of GroEL:GroES2 complexes allows the folding of two polypeptide molecules per GroEL reaction cycle, one in each ring of GroEL. By making use of a procedure to stabilize chaperonin complexes by rapid crosslinking for subsequent analysis by native PAGE, we have quantified the occurrence of GroEL:GroES1 and GroEL:GroES2 complexes in active refolding reactions under a variety of conditions using mitochondrial malate dehydrogenase (mMDH) as a substrate. Our results show that the symmetrical complexes are neither required for chaperonin function nor does their presence significantly increase the rate of mMDH refolding. In contrast, chaperonin-assisted folding is strictly dependent on the formation of asymmetrical GroEL:GroES1 complexes. These findings support the view that GroEL:GroES2 complexes have no essential role in the chaperonin mechanism. PMID- 10384960 TI - Regulation of cathepsin B activity by cysteine and related thiols. AB - We studied the mode of regulation of the activity of mature cathepsin B (CB) by L cysteine and some related thiols. The activity of CB with Z-Arg-Arg-NHMec as substrate was gradually inhibited over a range of increasing concentration of Cys, Cys methyl ester (CysOMe), Cys ethyl ester (CysOEt), N-acetyl-Cys (N-AcCys) and 3-mercaptopropionic acid. However, the inhibition of CB peaked at a definite value of [Cys], [CysOMe], [CysOEt] and [N-AcCys] and was gradually reversed over a range of higher concentrations of Cys and its esters. The maximum inhibitory concentrations of Cys, CysOME, CysOEt and N-AcCys showed a positive relationship to the pKa(RSH) values of the thiols and those of CysOEt and Cys decreased with increasing pH. The capability of the thiols to overcome their own inhibitory effect on CB was dependent on the concentration of their thiolate anion (RS-). However, the preincubation-dilution experiments showed that Cys and N-AcCys did not interact with active CB via a covalent mode. The inhibition of CB by N-AcCys was competitive and could be reversed by CysOMe. This activity-recovering effect of CysOMe was concentration-dependent and obeyed the Michaelis-Menten saturation kinetics over a profound increase of [RS-]. CB reacting in an environment of concurrently decreasing [RS-] and increasing [RSH], which was achieved by means of carboxylesterase-catalyzed deesterification of CysOEt to Cys, was progressively inhibited. Cys and N-AcCys also inhibited the fragmentation of histone H4 by CB and their concentration-dependent inhibitory profiles were qualitatively similar to those observed with Z-Arg-Arg-NHMec. Taken together, the results indicate that the RSH form of Cys and related thiols inhibits the activity of CB while the RS- form of these thiols counteracts or reverses the inhibitory action of the RSH form. This previously unrecognized thiol-thiolate anion regulation mechanism might be involved in a dynamic regulation of CB activity in endosomes and lysosomes and at the sites of lysosome-driven pericellular proteolysis. PMID- 10384961 TI - Epitope mapping of the monoclonal antibody MM12.10 to external MDR1 P glycoprotein domain by synthetic peptide scanning and phage display technologies. AB - Epitope mapping of MDR1-P-glycoprotein using specific monoclonal antibodies (mAbs) may help in delineating P-glycoprotein topology and hence in elucidating the relationship between its structural organization and drug-efflux pump function. In this work, by using synthetic peptide scanning and phage display technologies, the binding sites of the mAb MM12.10, a novel antibody to intact human multidrug resistant (MDR) cells, were studied. The results we obtained confirm that two regions localized on the predicted fourth and sixth loops are indeed external and that MDR1 peptides covering the inner domain of the current 12 transmembrane segment (TMs) model of P-glycoprotein could form part of the MM12.10 epitope. PMID- 10384962 TI - Molecular mimicry between a monoclonal antibody and one subunit of crotoxin, a heterodimeric phospholipase A2 neurotoxin. AB - Crotoxin is a heterodimeric phospholipase A2 neurotoxin formed by the non covalent association of an acidic and non-toxic subunit, CA, and a basic and weakly toxic phospholipase A2, CB. The two subunits behave in a synergistic manner. CA enhances the lethal potency of CB by increasing its selectivity of action. The mAb A-56.36, directed against the non-toxic subunit CA, was previously shown to neutralize crotoxin toxicity by dissociating the crotoxin complex. In the present report, a polypeptide sequence similarity was observed between some CDRs of mAb A-56.36 and two regions of CB (pos. 60-80 and 95-110). Phage displayed peptides corresponding to VH2 and VH3 of mAb A-56.36 and to their homologous sequences in CB bind CA to different extents. This observation shows that mAb A-56.36 interacts with a region of CA involved in its interaction with CB, therefore mimicking the binding of CB to CA. A similar approach was used to determine the regions of ammodytoxin A and of agkistrodotoxin, two phospholipase A2 neurotoxins similar to CB, which are involved in the formation of heterocomplexes with CA. The analysis of these data contributes to the determination of stretches of amino acids which could constitute the paratope of mAb A-56.36, as well as the region of association of CB with CA in crotoxin. PMID- 10384963 TI - Enzyme-linked immunosorbent assay for the measurement of JNK activity in cell extracts. AB - A colorimetric enzyme-linked immunosorbent assay (ELISA) for the measurement of kinase activity of c-Jun N-terminal kinases (JNKs) in cell extracts is described. The assay involves passive immobilisation of the substrate GST-cJun on the surface of a microtiter plate, selection of JNK protein kinases directly in substrate-coated wells, kinase reaction, and detection of substrate phosphorylation by a phosphoepitope-specific antibody. The ability of this assay to selectively measure JNK activity relies on the high-affinity interaction between JNKs and c-Jun. Accordingly, we found that JNKs could be captured on the microtiter plate surface through binding to the immobilised GST-cJun. Moreover, JNKs retained the specificity of their interaction with and phosphorylation of c Jun with respect to the dependence on both intact docking domain and the dimerisation state of c-Jun. This novel procedure represents a marked improvement on conventional radioactive assays in terms of sensitivity, accuracy of evaluation, low time consumption, high throughput and amenability to automation. It is expected to be useful forthe acceleration and facilitation of JNK activity measurement in cell extracts, in particular for large-scale screening of clinical samples. PMID- 10384964 TI - Mitochondrial DNA acts as potential promoter of the baculovirus RNA polymerase. AB - We have examined whether mitochondrial DNA could act as target of the RNA polymerase encoded by the baculovirus Autographa californica multicapsid nuclear polyhedrosis virus, because the baculovirus late promoters and the control region of host mitochondrial DNA show a high degree of sequence similarity. In vitro transcription using mitochondrial DNA from Spodoptera frugiperda cells and nuclear extracts prepared from baculovirus infected cells demonstrates that mitochondrial DNA is recognized by the viral RNA polymerase. Transcriptional initiation occurs at TAAG sequences, although not all of the six TAAG motifs present in the mitochondrial DNA fragment are recognized. The TAAG motif in the control region served as weak transcriptional start site, but some of the TAAG motifs in the coding sequences of the adjacent tRNA and rRNA genes are recognized efficiently. The sequences flanking the TAAG motifs used as transcriptional start sites have a lower helix stability than the flanking sequences of the nonfunctional TAAG motifs. These results support the view that helix stability rather than sequence specificity is an important factor for recognition of TAAG motifs by the viral RNA polymerase. PMID- 10384965 TI - In vitro phosphorylation of purified glycosylphosphatidylinositol-specific phospholipase D. AB - Glycosylphosphatidylinositol-specific phospholipase D (GPI-PLD) was phosphorylated in vitro by cAMP-dependent protein kinase (PKA) and by tyrosine kinase. Phosphorylation by PKA occurred in the 110 kDa native form of GPI-PLD as well as in multiple proteolytic degradation products and caused a significant decrease in enzyme activity. Dephosphorylation by treatment with alkaline phosphatase completely restored GPI-PLD activity. In addition, incubation of GPI PLD with trypsin, which results in the generation of distinct peptide fragments, resulted in complete dephosphorylation of radiolabeled GPI-PLD. The site of phosphorylation by PKA was assigned to Thr-286. Tyrosine phosphorylation was only observed in a proteolytically processed fragment of GPI-PLD but not in the 110 kDa native form and had no effect on GPI-PLD activity. PMID- 10384966 TI - Cathepsin S and cruzipain are inhibited by equistatin from Actinia equina. AB - Cathepsin S has been isolated for the first time from human tissue. It has a molecular mass of 24 kDa and an isoelectric point in the range of 8.2 to 8.6. The enzyme is inhibited by equistatin, which belongs to the thyropins, a new family of protein inhibitors, with an inhibition constant of Ki = 0.40 +/- 0.07 nM. Cruzipain, a cathepsin L-like enzyme sharing a 130 amino acid long C-terminal extension, is also strongly inhibited by equistatin (Ki = 0.028 +/- 0.006 nM). Together with previously reported data, these results further indicate that a functional heterogeneity exists among thyropin inhibitors, as demonstrated by their interaction with cathepsin S and cruzipain. PMID- 10384967 TI - Lipopeptides as dimerization inhibitors of HIV-1 protease. AB - In AIDS therapy, attempts have been made to inhibit the virus-encoded enzymes, e.g. HIV-1 protease, using active site-directed inhibitors. This approach is questionable, however, due to virus mutations and the high toxicity of the drugs. An alternative method to inhibit the dimeric HIV protease is the targeting of the interface region of the protease subunits in order to prevent subunit dimerization and enzyme activity. This approach should be less prone to inactivation by mutation. A list of improved 'dimerization inhibitors' of HIV-1 protease is presented. The main structural features are a short 'interface' peptide segment, including non-natural amino acids, and an aliphatic N-terminal blocking group. The high inhibitory power of some of the lipopeptides [e.g. palmitoyl-Tyr-Glu-Leu-OH, palmitoyl-Tyr-Glu-(L-thyronine)-OH, palmitoyl-Tyr-Glu (L-biphenyl-alanine)-OH] with low nanomolar Ki values in the enzyme test suggests that mimetics with good bio-availability can be derived for AIDS therapy. PMID- 10384968 TI - What's in a name? PMID- 10384969 TI - Using flavoxate as primary medication for patients suffering from urge symptomatology. AB - A drug utilization observation study collected data on a total of 1800 patients given flavoxate (Spasuret 200) over 2 weeks for urge incontinence. Efficacy and tolerance parameters were determined. A subgroup of 618 patients without urinary tract infections or benign prostatic hyperplasia were treated with flavoxate only. The subgroup (n = 618) showed a reduction of dysuria (37%), nocturia (53%), and both daytime (61%) and nighttime urge (69%). Bladder volume at first urge sensation increased by 55.1+/-58.8 ml (36%), which was comparable to data from the entire group (1800 patients). In 89.2% of all patients the residual urine volume was stable or decreased. Undesirable side effects occurred in 1.8% of cases. Both groups showed better results with flavoxate four times daily (800 mg), compared to three times daily (600 mg). Flavoxate is effective and well tolerated and causes no additional problems due to residual urine or side effects. PMID- 10384970 TI - The anatomic and functional variability of rectoceles in women. AB - Fluoroscopic parameters of the rectum in women with pelvic organ prolapse were studied. Ninety-eight consecutive women undergoing reconstructive pelvic surgery completed a urogynecologic history with physical examination and pelvic floor fluoroscopy. The presence of rectocele and contrast trapping was determined on each fluoroscopic study. Each frame of the study was measured to determine the rectal width. Seventy-eight per cent of the women had fluoroscopically demonstrated rectoceles. Their maximum and minimum rectal widths were larger than those of women without rectoceles. Contrast-retaining rectoceles were larger than non-contrast retaining rectoceles. Fluoroscopic evidence of contrast retention did not relate to patient symptoms. There was no difference in the grade of posterior wall prolapse in women with and without rectoceles. Rectoceles have anatomic and functional variability. Fluoroscopy may be a valuable adjunct to the physical examination in assisting gynecologic surgeons to refine their surgical approach for rectocele repair. PMID- 10384971 TI - Effect of a urinary control insert on quality of life in incontinent women. AB - The aim of the study was to evaluate the efficacy, safety and effect on quality of life of the Reliance urinary control insert (Uromed Corp., Needham, MA) in women with genuine stress incontinence. Efficacy was evaluated at baseline and at the end of the 12-month study period by standardized pad-weight studies and by rating scales measuring acceptability, incontinence symptom improvement, ease of learning, comfort and time to habituation, recorded in diaries at monthly intervals in 63 women. The SF-36 Health Survey questionnaire was used to assess quality of life status at baseline without the device and after 12 months of device use. A significant decrease in urine loss at 12 months compared with baseline was shown by standardized pad-weight studies, with and without the device in situ. Urine loss was reduced by more than 80% in 91% of the 63 patients, and 79% were completely dry. Patient diaries showed significant improvement in control of leakage, comfort, and ease of device use during the study period. Short-term-36 Health Status data also indicated significant improvement in the physical functioning score at 12 months. Urinary tract infection and hematuria were the most common adverse effects. The Reliance urinary control insert is an efficacious and safe means of controlling genuine stress incontinence in women. The device was perceived as easy to use and comfortable for these 63 women, and resulted in improved quality of life. PMID- 10384972 TI - Pharmacologic causes of female incontinence. AB - The etiology of female urinary incontinence is complex and multifactorial. Many medications have adverse effects on the lower urinary tract, including the promotion of incontinence in certain women. Medications may cause incontinence through three main mechanisms: decreased intraurethral pressure, increased intravesical pressure, and indirect effects on the lower urinary tract. Careful adjustments of a patient's medications based on a knowledge of pharmacologic mechanisms of action may restore continence in some women. PMID- 10384973 TI - Pad testing in incontinent women: a review. AB - This article reviews the literature on pad-weighing tests used for objectifying and quantifying incontinence in urinary incontinent women. The patients wear pads weighed before and after the test period. A weight gain is taken as a measure of the amount of urine loss. The tests are in principle of two different types: short-term office tests and long-term home tests, and measure different aspects of urinary control and dysfunction. Both have an inherent large intra- and interindividual variability. Pad weight gains obtained from patients referred for incontinence and those from self-reported continent controls overlap to a certain degree, and it is not possible to identify distinct numerical cut-off values separating continence from incontinence. This suggests that incontinence is a complex condition in which the amount of leakage, other sources of weight gain, and differences in the individual patients' personal characteristics influence the identification and quantification of the problem. In spite of the shortcomings the pad tests remain a valuable tool for both the clinician and the researcher. The home pad tests are superior to the office tests in terms of authenticity, and should be performed with a concomitant systematic registration of the participant's voidings, fluid intake and episodes of incontinence. PMID- 10384974 TI - Laparoscopic management of recurrent vesicovaginal fistula. AB - Vesicovaginal fistula repair is most commonly undertaken via a transvaginal approach. We report a recurrent case of vesicovaginal fistula which was ultimately repaired using a laparoscopic approach. The fistula followed a hysterectomy and persisted despite two operations using the Latzko partial colpocleisis and prolonged catheterization. The fistulous tract was ultimately repaired by closing the vagina and bladder with an interposing omental flap utilizing a laparoscopic approach. PMID- 10384975 TI - Evaluation of the female with neurogenic voiding dysfunction. AB - Neurological disease can have a profound effect on the lower and upper urinary tracts. Symptoms, such as incontinence and inability to void, as well as sequelae of hydronephrosis and renal insufficiency, are not uncommon. It is imperative that neurogenic voiding dysfunction by aggressively treated to avoid these problems. Care of the patient with neurogenic voiding dysfunction starts with an understanding of the neurophysiology of micturition and how this can be affected by various neurological diseases. The clinician must also be able to classify voiding dysfunction through history and physical examination and urodynamic testing so that proper treatment can be instituted. PMID- 10384976 TI - Voiding dysfunction in women with diabetes mellitus. AB - Voiding dysfunction is a common finding in women with diabetes mellitus. A full spectrum of bladder dysfunction, from areflexia to detrusor instability, can result. Urodynamics is frequently required to identify the specific disorder. Current concepts regarding the evaluation and treatment of these patients are reviewed. PMID- 10384977 TI - Voiding dysfunction in women with lumbar disc prolapse. AB - A significant proportion of women with lumbar disc prolapse experience voiding dysfunction. The most common finding is detrusor areflexia, frequently associated with impaired sensation. The pertinent neuropathophysiologic findings, clinical features and methods of evaluation and treatment are reviewed. PMID- 10384978 TI - Evaluation and management of lower urinary tract disorders in women with multiple sclerosis. AB - This paper outlines the evaluation and management of the lower urinary tract abnormalities related to voiding function in women with multiple sclerosis (MS). For the pelvic floor reconstructive surgeon, it is important to realize that every patient with MS may have voiding dysfunction unrelated to lower urinary tract symptoms, duration of disease or disability status. Proper evaluation and individualized management of the urinary tract reduces the morbidity and improves the quality of life of patients with this degenerative neurologic disorder. PMID- 10384979 TI - Female voiding dysfunction and movement disorders. AB - Voiding dysfunction associated with Parkinson's disease has been well described in male patients. Few studies detail voiding dysfunction in female Parkinson patients. Apparent differences between patients with Parkinson's subtypes have also not been sufficiently defined. The majority of female Parkinson patients who have urinary symptoms (>70%) will manifest symptomatic urgency with or without urge incontinence. The remaining patients will have mixed irritative and obstructive or purely obstructive symptoms. Urodynamic evaluation demonstrates detrusor hyperreflexia in 70%-80% of female patients. However, women with Parkinson-related syndromes demonstrate detrusor hypocontractility or areflexia in 20%-30% of cases. Electromyography reveals sphincteric dysfunction (pseudodyssynergia, bradykinesia) in 30%-50% of female Parkinson patients. Also, in patients with Parkinson-related syndromes a high prevalence of peripheral denervation can be documented on electromyographic study of the pelvic floor. Voiding dysfunction associated with Parkinson's disease in female patients is complex and not always congruent with symptoms. Urodynamic evaluation is crucial to fully elucidate lower urinary tract dysfunction in female patients with Parkinson's and Parkinson-related disorders. PMID- 10384980 TI - Localization of a centrin-like protein to higher plant plasmodesmata. AB - Antibodies against centrin, the ubiquitous calcium-binding contractile protein, recognized a 17 kDa protein in extracts of onion root tips and cauliflower florets. Using immunofluorescence microscopy, anti-centrin antibodies were localized to the developing cell plate of onion and cauliflower root tip cells. In cauliflower florets, these antibodies localized to the walls in a punctate manner, consistent with the distribution of plasmodesmata as shown by colocalization with callose. Anti-centrin antibodies were localized to plasmodesmata of onion root tips and cauliflower florets with immunogold electron microscopy. Furthermore, this label was concentrated around the necks of plasmodesmata. In contrast, an antibody against calmodulin, which is a closely related calcium-binding protein, did not label plasmodesmata. We propose that centrin is a component of calcium-sensitive contractile nanofilaments in the neck region of plasmodesmata and facilitates the calcium-induced regulation of intercellular transport. PMID- 10384981 TI - The ADP-ribosylation factor GTPase-activating protein Glo3p is involved in ER retrieval. AB - Retrograde transport of proteins from the Golgi to the endoplasmic reticulum (ER) has been the subject of some interest in the recent past. Here a new thermosensitive yeast mutant defective in retrieval of dilysine-tagged proteins from the Golgi back to the endoplasmic reticulum was characterized. The ret4-1 mutant also exhibited a selective defect in forward ER-to-Golgi transport of some secreted proteins at the non-permissive temperature. The corresponding RET4 gene was found to encode Glo3p, a GTPase-activating protein (GAP) specific for ADP ribosylation factor (ARF). In vitro, the Glo3 thermosensitive mutant showed a reduced ARF1-GAP activity. The Glo3 protein belongs to a family of zinc finger proteins that may include additional ARF-GAPs. Gene deletion experiments of other family members showed that only GLO3 deletion resulted in impaired retrieval of dilysine-tagged proteins back to the ER. These results demonstrate that Glo3p is the main ARF-GAP specifically involved in ER retrieval. PMID- 10384982 TI - Differential distribution of G-protein beta-subunits in brain: an immunocytochemical analysis. AB - Heterotrimeric G proteins play central roles in signal transduction of neurons and other cells. The variety of their alpha-, beta-, and gamma-subunits allows numerous combinations thereby confering specificity to receptor-G-protein effector interactions. Using antisera against individual G-protein beta-subunits we here present a regional and subcellular distribution of Gbeta1, Gbeta2, and Gbeta5 in rat brain. Immunocytochemical specificity of the subtype-specific antisera is revealed in Sf9 cells infected with various G-protein beta-subunits. Since Gbeta-subunits together with a G-protein gamma-subunit affect signal cascades we include a distribution of the neuron-specific Ggamma2- and Ggamma3 subunits in selected brain areas. Gbeta1, Gbeta2, and Gbeta5 are preferentially distributed in the neuropil of hippocampus, cerebellum and spinal cord. Gbeta2 is highly concentrated in the mossy fibres of dentate gyrus neurons ending in the stratum lucidum of hippocampal CA3-area. High amounts of Gbeta2 also occur in interneurons innervating spinal cord alpha-motoneurons. Gbeta5 is differentially distributed in all brain areas studied. It is found in the pyramidal cells of hippocampal CA1-CA3 as well as in the granule cell layer of dentate gyrus and in some interneurons. In the spinal cord Gbeta5 in contrast to Gbeta2 concentrates around alpha-motoneurons. In cultivated mouse hippocampal and hypothalamic neurons Gbeta2 and Gbeta5 are found in different subcellular compartments. Whereas Gbeta5 is restricted to the perikarya, Gbeta2 is also found in processes and synaptic contacts where it partially colocalizes with the synaptic vesicle protein synaptobrevin. An antiserum recognizing Ggamma2 and Ggamma3 reveals that these subunits are less expressed in hippocampus and cerebellum. Presumably this antiserum specifically recognizes Ggamma2 and Ggamma3 in combinations with certain G alphas and/or Gbetas. The widespread but regionally and cellularly rather different distribution of Gbeta- and Ggamma2/3-subunits suggests that region-specific combinations of G-protein subunits mediate signal transduction in the central nervous system. The different subcellular distribution of Gbeta subunits in cultivated neurons reflects that observed in tissue where Gbeta5 and Gbeta2 associate preferentially with the perikarya and the neuropil, respectively, and suggests an additional association of Gbeta2 with secretory vesicles. PMID- 10384983 TI - A regulatory role of fibroblast growth factor in the expression of decorin, biglycan, betaglycan and syndecan in osteoblasts from patients with Crouzon's syndrome. AB - Bone development is controlled by the autocrine and/or paracrine effects of regulatory molecules. We previously showed that the phenotype of fibroblasts obtained from patients affected by Crouzon's syndrome, an autosomal dominant disease characterized by pathological skull bone development, differed from that of normal cells and was regulated by interleukin treatments. The changes in the relative concentrations of extracellular macromolecules (glycosaminoglycans-GAG, collagen and fibronectin) were associated with abnormal interleukin secretion that affected the microenvironment where the osteogenic processes take place. Mutations in human fibroblast growth factor receptors are now thought to be involved in Crouzon's syndrome. Since coactivation of interleukins and basic fibroblast growth factor (bFGF) is probably implicated in morphogenetic and osteogenic processes and heparan sulphate proteoglycans have a critical role in regulating bFGF activity, the phenotypes of normal and Crouzon osteoblasts were studied and the effects of bFGF on the expression of bFGF, procollagen alpha1 (I), and proteoglycan (PG) genes for biglycan, decorin, betaglycan and syndecan analyzed. Specific human cDNA probes were used to screen the relative levels of mRNA by Northern analysis. Spontaneous or bFGF-modulated release of interleukins was also assayed. The bFGF gene transcript was detected only in Crouzon osteoblasts. We showed for the first time that Crouzon osteoblasts, despite a mutation in the FGF receptor, still responded to exogenous bFGE In fact, the growth factor induced changes in the GAG profile and in the levels of mRNA coding for PG and procollagen alpha1 (I) and down-regulated heparan sulfate GAG chains. ELISA showed that bFGF-induced interleukin secretion differed in normal and Crouzon osteoblasts. The observed differences in PG core protein, procollagen alpha1 (I) and bFGF could be associated with the Crouzon bone phenotype and also should provide further understanding on the molecular basis of the diseased state of bone. PMID- 10384984 TI - 45T-1, an established cell line with characteristics of Sertoli cells, forms organized aggregates in vitro after exposure to tumor necrosis factor alpha. AB - In the testis TNF is produced by germinal cells. The putative role of tumor necrosis factor alpha (TNF) in development and differentiation was investigated in 45T-1 mouse cell cultures, a cell line with characteristic markers of Sertoli cells, established from transgenic mouse families expressing the polyoma large T antigen in their testes. Exposure to TNF elicited a gradual assembly of the cells of the monolayer into highly organized spheroids. The first morphological sign of the changes was detected one week after TNF treatment by anti-desmin immunostaining which showed the formation of foci in the culture consisting of several hundred cells connected by an increasing number of cell contacts. Between days 10-20 the cells formed large ovoid or vermiform aggregates covered by several layers of flat, elongated cells. These cells extended septae into the inner mass of the spheroids consisting of loosely arranged, large polygonal or palisadic cells. The spheroids were surrounded by radially arranged elongated cells covered by small blebs. TNF treatment upregulated laminin expression in 45T 1 cell cultures, which is known to induce formation of cord-like structures by Sertoli cells in vitro. Coculturing 45T-1 cells with immortalized germinal cells or TNF-producing HeLa cells also lead to the formation of spheroids. These observations suggest that TNF production of germinal cells might contribute to the organization/differentiation of Sertoli cells. PMID- 10384985 TI - Immunological analyses of alkyl-dihydroxyacetone-phosphate synthase in human peroxisomal disorders. AB - Alkyl-dihydroxyacetonephosphate synthase (alkyl-DHAP synthase) is a peroxisomal enzyme involved in the biosynthesis of ether phospholipids. To localize the enzyme in human peroxisomal disorders, indirect immunofluorescence and immunoblot analysis was performed. In Zellweger syndrome and rhizomelic chondrodysplasia punctata fibroblast cell lines, alkyl-DHAP synthase protein levels on immunoblots were strongly decreased and residual immunofluorescence was diffusely localized throughout the cytoplasm. In a particular neonatal adrenoleukodystrophy cell line, characterized by the absence of a functional peroxisomal targeting signal 1 receptor, the precursor form of the enzyme was detected in Western blots at levels comparable to that of the mature enzyme in control fibroblasts. Similarly, fibroblasts from patients with a single deficiency in the activity of either alkyl-DHAP synthase or DHAP-acyltransferase showed normal levels of the mature alkyl-DHAP synthase protein on immunoblots. Immunofluorescence experiments revealed a peroxisomal localization of both the precursor and the mature form of the enzyme. Collectively, these results visualize the peroxisomal localization of alkyl-DHAP synthase, indicate that the enzyme is unstable outside its target organelle and explain that normal enzyme protein levels found in some peroxisomal disorders result from protection against cytoplasmic degradation through import into peroxisomes. Additionally, alkyl-DHAP synthase could be detected in rat mesangial cells and murine NIH-3R3 fibroblasts by immunofluorescence as well as immunoblot analysis. Immunoelectron microscopy showed that the enzyme is predominantly located on the lumenal side of the peroxisomal membrane in rat and guinea pig liver. PMID- 10384986 TI - Highly divergent amino termini of the homologous human ALR and yeast scERV1 gene products define species specific differences in cellular localization. AB - The yeast scERV1 gene product is involved in the biogenesis of mitochondria and is indispensable for viability and regulation of the cell cycle. Recently the general importance of this gene for the eukaryotic cell was shown by the identification of a structural and functional human homologue. The homologous mammalian ALR (Augmenter of Liver Regeneration) genes from man, mouse and rat are involved in the phenomenon of liver regeneration. A low expression rate of the genes is found in all investigated cells and mammalian tissues but it is specifically induced after damage of liver organs and is especially high during spermatogenesis. The alignment of the different proteins identifies a highly conserved carboxy terminus with more than 40% identical amino acids between yeast and mammals. The conserved carboxy terminus is functionally interchangeable between distantly related species like yeast and man. In contrast, the amino terminal parts of the proteins display a high degree of variability and significant differences even among closely related species. This finding leads to the problem whether the amino termini have comparable or divergent functions in different species. In this study we demonstrate by heterologous complementation experiments in yeast that the complete human ALR protein with its own amino terminus is not able to substitute for the yeast scERV1 protein. Fusion proteins of Alrp and scErv1p with the green fluorescence protein were created to investigate the respective subcellular localizations of these homologous proteins in yeast and human cells. In yeast cells human Alrp accumulates in the cytoplasm in contrast to yeast scErv1p that is preferentially associated with yeast mitochondria. Comparable studies with human cells clearly show that the homologous human Alrp is located in the cytosol of these cells. Fractionation experiments and antibody tests with yeast and human mitochondria and cellular extracts verify these findings. PMID- 10384987 TI - Tumors of peripheral nerves: correlation of symptoms, clinical signs, imaging features, and histologic diagnosis. AB - OBJECTIVE: To distinguish between benign and malignant tumors in the peripheral nerves. DESIGN AND PATIENTS: The clinical, imaging and histologic findings of 99 benign and 16 malignant tumors in the peripheral nerves were reviewed retrospectively. RESULTS: Preoperative motor weakness was observed in only six of 99 benign tumors and was mild, while slight to severe motor weakness was present in 15 of 16 malignant lesions. Pain at rest was present in five of 99 benign tumors and in 15 of 16 malignant tumors. All benign lesions showed a smooth tumoral margin, while half the malignant lesions showed an invasive margin on CT or MRI. Thirteen of 28 benign lesions on CT and nine of 23 on MRI showed round to geographic central enhancement, but this pattern was not seen in malignant lesions. CONCLUSION: Absence of severe motor weakness and a central enhancement pattern strongly suggest a benign nature, while severe rest pain and invasive tumor margin suggest malignant lesions in peripheral nerve tumors. PMID- 10384988 TI - Variations in meniscofemoral ligaments at anatomical study and MR imaging. AB - PURPOSE: To demonstrate variations in the meniscofemoral ligaments (ligaments of Wrisberg and Humphrey) at anatomical study and magnetic resonance (MR) imaging. DESIGN: Twenty-eight cadaveric knees were partially dissected for the examination of the meniscofemoral ligaments. One hundred knee MR examinations were reviewed by two experienced musculoskeletal radiologists. Proximal variations in the meniscofemoral ligaments at MR imaging were classified into three types according to the attachment site: type I, medial femoral condyle; type II, proximal half of the posterior cruciate ligament (PCL); type III, distal half of the PCL. Distal variations were classified into vertical or oblique types according to the orientation of the intermediate signal at the interface of the ligament and lateral meniscus. RESULTS: At anatomical study, six cases showed variations in the proximal insertion site of the meniscofemoral ligaments. At MR imaging 93 cases had one or more meniscofemoral ligaments, giving a total of 107 ligaments: 90 ligaments of Wrisberg and 17 ligaments of Humphrey. Forty-one ligaments of Wrisberg were type I, 28 type II, 19 type III, and with two indeterminate type, while 6 ligaments of Humphrey were type I and the remaining 11 were indeterminate. Seven cases showed no meniscofemoral ligament. Of the 107 meniscofemoral ligaments, the distal insertion orientation was of vertical type in 10 ligaments, oblique type in 70 and unidentified in 27. CONCLUSION: An understanding of the high incidence of meniscofemoral ligament variations may help in the interpretation of knee MR studies. PMID- 10384989 TI - The prevalence of sacroilitis in psoriatic arthritis: new perspectives from a large, multicenter cohort. A Department of Veterans Affairs Cooperative Study. AB - OBJECTIVE: To determine the prevalence of radiographic evidence of sacroiliitis in a large population of patients with psoriatic arthritis. PATIENTS AND DESIGN: Patients were recruited from 15 clinical centers. This was part of a large, multicenter study of patients with an established diagnosis of ankylosing spondylitis, psoriatic arthritis, or reactive arthritis. For this cohort, an established diagnosis of psoriatic arthritis was required, with cutaneous manifestations and involvement of at least three appendicular joints. At entry, patients were not selected for the presence of axial involvement. Radiographs - one anteroposterior view of the pelvis and one oblique view of each sacroiliac joint - were graded using the New York classification scale by a musculoskeletal radiologist masked to the specific diagnosis and clinical symptoms. Re-evaluation of 10% of the films 3 years later quantified intraobserver variability. RESULTS: Two hundred and two patients with psoriatic arthritis were studied. Duration of the disease averaged 12 years; all patients had psoriasis and peripheral arthritis. The prevalence of radiographic evidence of sacroiliitis (grade 2 or higher) was 78%; 71% of these had grade 3 disease. CONCLUSIONS: Previously reported prevalence of sacroiliitis in patients with psoriatic arthritis ranges from 30% to 50%. The prevalence of radiographic evidence of sacroiliitis in this large multicenter cohort of patients with appendicular psoriatic arthritis was substantially higher. PMID- 10384990 TI - Post-traumatic and stress-induced osteolysis of the distal clavicle: MR imaging findings in 17 patients. AB - OBJECTIVE: To describe the MR imaging findings in patients with osteolysis of the distal clavicle and to compare the MR imaging appearance of clavicular osteolysis following acute injury with that related to chronic stress. DESIGN AND PATIENTS: MR imaging examinations were reviewed in 17 patients (14 men, 3 women; ages 16-55 years) with the diagnosis of post-traumatic or stress-induced osteolysis of the clavicle. A history of a single direct injury was present in seven patients and a history of weight-lifting, participation in sports, or repetitive microtrauma was present in 10 patients. RESULTS: MR imaging showed edema in the distal clavicle in 17 patients and, of these, eight also had edema in the acromion. The edema was most evident in STIR and fat-suppressed T2-weighted pulse sequences. Other findings about the acromioclavicular (AC) joint were prominence of the joint capsule in 14, joint fluid in eight, cortical irregularity in 12, and bone fragmentation in six patients. No differences in the MR imaging features of post traumatic and stress-induced osteolysis of the distal clavicle were observed. CONCLUSION: Post-traumatic and stress-induced osteolysis of the distal clavicle have similar appearances on MR imaging, the most common and conspicuous MR imaging feature being increased T2 signal intensity in the distal clavicle. PMID- 10384991 TI - Peripheral focal low signal intensity areas in the degenerated annulus fibrosus on T2-weighted fast spin echo MR images: correlation with macroscopic and microscopic findings in elderly cadavers. AB - OBJECTIVE: To correlate the peripheral focal low signal intensity areas in the degenerated annulus fibrosus on T2-weighted fast spin echo MR images with the macroscopic and microscopic findings in cadavers derived from elderly subjects. DESIGN: Twenty-eight intervertebral disks (16 lumbar and 12 cervical) derived from four nonembalmed cadavers were examined with T1-weighted spin echo and proton density-weighted and T2-weighted fast spin echo MR imaging. The signal intensities of the annulus fibrosus were evaluated on sagittal MR images and correlated with the findings on corresponding sagittal anatomic sections. The MR imaging-histologic correlation was then studied. RESULTS: Peripheral focal low signal intensity areas and adjacent regions of high signal intensity were found in five lumbar intervertebral disks. Peripheral focal low signal intensity regions consisted of disorganized compact annular fibers, tiny fissures, and dense fibrosis. The high signal intensity regions, adjacent to the areas of low signal intensity, consisted of mucoid degeneration, tiny fissures, and chondroid metaplasia. CONCLUSIONS: Awareness of the histologic findings in regions that reveal peripheral focal low signal intensity with adjacent regions of high signal intensity in the degenerated annulus fibrosus on T2-weighted images may facilitate effective interpretation of clinical MR images of the spine. PMID- 10384992 TI - MR imaging and CT in osteoarthritis of the lumbar facet joints. AB - OBJECTIVE: To test the agreement between MR imaging and CT in the assessment of osteoarthritis of the lumbar facet joints, and thus to provide data about the need for an additional CT scan in the presence of an MR examination. DESIGN AND PATIENTS: Using a four-point scale, two musculoskeletal radiologists independently graded the severity of osteoarthritis of 308 lumbar facet joints on axial T2-weighted and on sagittal T1- and T2-weighted turbo-spin-echo images and separately on the corresponding axial CT scans. Kappa statistics and percentage agreement were calculated. RESULTS: The weighted kappa coefficients for MR imaging versus CT were 0.61 and 0.49 for readers 1 and 2, respectively. The weighted kappa coefficients for interobserver agreement were 0.41 for MR imaging and 0.60 for CT, respectively. There was agreement within one grade between MR and CT images in 95% of cases for reader 1, and in 97% of cases for reader 2. CONCLUSION: With regard to osteoarthritis of the lumbar facet joints there is moderate to good agreement between MR imaging and CT. When differences of one grade are disregarded agreement is even excellent. Therefore, in the presence of an MR examination CT is not required for the assessment of facet joint degeneration. PMID- 10384993 TI - Proximal patellar tendinosis and abnormalities of patellar tracking. AB - OBJECTIVE: To assess whether an association exists between patellar tendinosis and abnormal patellar tracking. DESIGN AND PATIENTS: The MRI examinations of 630 patients (i.e. 860 knees) referred with anterior knee pain over a 4-year period were assessed in retrospect for the presence of patellar tendinosis and abnormal patellar tracking. The images of the patients with patellar tendinosis were reviewed and the location within the patellar tendon was recorded. RESULTS: There were 44 knees with proximal patellar tendinosis. Twenty-four of these were considered to have normal patellar tracking and 20 to have abnormal patellar tracking. In the group of 816 knees without proximal patellar tendinosis, 581 were considered to have normal patellar tracking and 235 knees to have abnormal patellar tracking. When the two groups were compared there was a statistically significant difference in the ratio of patients with and without abnormal tracking. CONCLUSION: In patients referred with anterior knee pain or suspected abnormal patellar tracking there is a significant association between proximal patellar tendinosis and abnormal patellar tracking. PMID- 10384994 TI - Hypertrophic osteoarthropathy of one leg--a sign of aortic graft infection. AB - We report a rare case of hypertrophic osteoarthropathy (HOA) confined to the right leg secondary to aortic graft infection. The development of HOA exclusively localized to areas distal to a vascular prosthesis may be the presenting manifestation of graft infection and a crucial diagnostic clue in the early detection of vascular graft infection. HOA is diagnosed by its characteristic radiographic and scintigraphic pattern. Most prosthetic, especially aortic, graft infections are uniformly fatal if not treated by aggressive surgical and antibiotic therapy. Recognition of this uncommon association may facilitate an early diagnosis, which usually requires immediate surgical therapy. PMID- 10384995 TI - Hypertrophy and pseudohypertrophy of the lower leg following chronic radiculopathy and neuropathy: imaging findings in two patients. AB - Enlargement of the ipsilateral muscle compartment is an exceptional finding in patients with chronic radiculopathy, peripheral nerve injury, anterior horn cell diseases, or acquired peripheral neuropathy. We report radiographic, ultrasonographic, CT and MRI findings in a patient with chronic S1 radiculopathy and another with chronic neuropathy of the common fibular nerve (L4-S2), both presenting with painless enlargement of the calf muscles. PMID- 10384996 TI - Prenatal diagnosis of distal arthrogryposis type 1. AB - A 23-year-old woman, gravida 1 parity 0, was referred for routine ultrasound examination at 23 weeks gestation. Fetal anatomy was normal. However, both hands demonstrated clasped thumbs without extension. Repeated ultrasound examination verified the bilateral hands posture. The position of the hands did not change following sound stimulation. The child was diagnosed as distal arthrogryposis type 1. Prenatal counselling by the pediatric orthopedic surgeon, geneticist and gynecologist, was provided, to inform the parents on the probable outcome of the fetus and the hands. The parents were advised to continue with the pregnancy. A 1,975-g healthy boy was delivered by spontaneous vaginal delivery. Neonatal examination confirmed the prenatal diagnosis of arthrogryposis type 1. Following reconstructive surgery the child functions well with both his hands. PMID- 10384997 TI - Fracture of the lateral process of the talus: appearance at MR imaging and clinical significance. AB - The case of a 59-year-old man with chronic lateral ankle pain following an inversion injury is presented. MR imaging performed to evaluate for soft tissue injury revealed an unsuspected fracture of the lateral process of the talus. The patient underwent surgical exploration of the fracture with debridement of adjacent loose bodies and is currently undergoing aggressive physical rehabilitation. PMID- 10384998 TI - Primary clear cell sarcoma of bone: a unique site of origin. AB - Clear cell sarcoma is a rare soft tissue neoplasm, accounting for less than 1% of soft tissue sarcomas. We are presenting a case of a clear cell sarcoma of bone which, to our - knowledge, is the only report of a , primary clear cell sarcoma of bone. PMID- 10384999 TI - Purification and characterisation of a minor low-sulphated dermatan sulphate proteoglycan from ray skin. AB - A minor low-sulphated dermatan sulphate proteoglycan was isolated from ray skin by extraction with 2% sodium dodecyl sulphate, followed with ion-exchange chromatography, gel chromatography and density gradient centrifugation. The proteoglycan with a relative molecular mass (Mr) ranging from 70 to 120 kDa is composed of about two dermatan sulphate chains (Mr 33 kDa) bound on a protein core of Mr 27 kDa, and oligosaccharides consisting of uronic acids, hexosamines and neutral sugars. The major amino acids of the protein core were glycine (corresponding to about one-fourth of the total amino acids), serine, threonine, glutamic acid/glutamine, leucine and cysteine, together amounting to 56% of the total. The isolated proteoglycan does not interact with hyaluronic acid and does not form self-aggregates. Dermatan sulphate was rich in iduronic acid (62% of total uronic acid) and composed of non-sulphated (44%), and mono-sulphated disaccharides bearing esterified sulphate groups at positions C-4 (53%) or C-6 (3%) of the N-acetyl galactosamine. HPLC analysis of a pure preparation of dermatan sulphate, showed the presence of galactose and glucose possibly as branches on the dermatan sulphate chain. PMID- 10385000 TI - Detection of an extended-10 element in the promoter region of the pckA gene encoding phosphoenolpyruvate carboxykinase in Escherichia coli. AB - The regulation of transcription of the pckA gene coding for phosphoenolpyruvate carboxykinase in Escherichia coli was analysed by site-directed mutagenesis of the promoter region and measurement of in vitro transcription initiation. Mutation of the guanine residue at position -14 to either cytosine or thymine was found to result in a drastic decrease of transcription, even in the presence of the natural -35 hexamer TTTCCA that differs by only two nucleotides from the consensus sequence TTGACA. It was concluded that the promoter region of pckA contains an extended -10 module, 5'-TG-3', located one base upstream of the -10 hexamer, which is crucial for transcription. PMID- 10385001 TI - A Y form of hammerhead ribozyme trapped by photo-cross-links retains full cleavage activity. AB - The conformation in solution of a small bipartite I-III hammerhead ribozyme has been deduced from the photo-crosslinks formed between cleavable ribo deoxysubstrates appropriately substituted with the probe deoxy-4-thiouridine and ribozyme residues. The ribozyme-substrate complex is able to adopt a Y-like structure with stems I and II in close proximity in the presence of 400 mM Na+ only. Indeed, a cross-link joining stem I (1.6) to loop II (AL2.4) forms in significant amount under these conditions. This cross-linked complex furthermore elicits, upon Mg2+ addition, a catalytic activity similar to that exhibited by the complexes cross-linked at the distal ends of either stem I or stem III or of the non-substituted bipartite complex. This shows that the reaction mechanism is fully compatible with a strong structural constraint between stems I and II and that sodium ions at high concentration (400 mM) are able to promote a proper folding of hammerhead ribozymes. None of the multiple cross-links formed within the ribozyme core (probe in position 16.1 or 1.1) was found catalytically active. The cross-link patterns nevertheless indicate a higher flexibility of the core in Na+ than in Mg2+. While most of the cross-links can be accommodated by the Y solution structure, some of them (16.1 to U4 and 2.1) definitely can not, suggesting that additional alternative inactive conformations exist in solution. PMID- 10385002 TI - A SECIS binding protein (SBP) is distinct from selenocysteyl-tRNA protecting factor (SePF). AB - In mammals, most of the selenium contained in their body is present as an unusual amino acid, selenocysteine (Sec), whose codon is UGA. Because the UGA codon is normally recognized as a translational stop signal, it is intriguing how cells recognize and distinguish the UGA Sec codon from the UGA stop codon. In eukaryotic selenoprotein mRNAs, it has been proposed that a conserved stem-loop structure designated Sec insertion sequence (SECIS) located in the 3' untranslated regions is required for recognition of UGA as a Sec codon. Although some proteins (SBPs) have been reported to bind to SECIS, it is not clear how the SECIS element can mediate Sec insertion at UGA. Eukaryotic Sec-tRNA(Sec) is not recognized by elongation factor EF-1alpha, but is recognized specifically by a Sec-tRNA(Sec) protecting factor, SePF, in bovine liver extracts. In this study, we provide evidence that SePF is distinct from SBP by chromatography. Upon UV irradiation, the SECIS RNA was cross-linked to a 47.5 kDa protein, a likely candidate of SBP, that is contained in the complex with a molecular mass of 150 kDa. These results suggest that SBP and SePF play different roles for the Sec incorporation. To our knowledge, this is the first demonstration that SBP is discriminated from the factor which directly recognizes Sec-tRNA(Sec), providing a novel clue to the mechanism of selenocysteine decoding in eukaryotes. PMID- 10385003 TI - Analysis of the human HLA-DRA gene upstream region: evidence for a stem-loop array directed by nuclear factors. AB - Sequence analysis of the far-upstream region of the human HLA-DRA gene has revealed the presence of Y' and X' boxes, highly homologous to the well characterized Y and X boxes present within the proximal-promoter region. Comparison of Y, Y', X, and X' box sequences present within different class II MHC genes of different species demonstrates that these boxes are conserved during evolution, suggesting an important role in regulation of gene expression. The far upstream region and the proximal promoter region of the class II MHC genes could be organized in secondary structures, as suggested for the EA gene, the murine counterpart of the human HLA-DRA gene. The essential feature of this model is a dimerization of the proteins binding to X and X' and/or Y and Y' boxes resulting in a loop-out of the intervening DNA and a rapprochement of the far-upstream and proximal-promoter regions, and consequently of any proteins binding to them. We set up an in vitro approach in order to determine whether proteins bound to sequences present within far-upstream and proximal-promoter regions of the human HLA-DRA gene could direct a secondary structure assembly of regulative regions. Moreover, by gel retardation and DNase I footprinting assays, we demonstrate that similar proteins bind to Y and Y' boxes and, among these proteins, NF-Y was unambiguously identified by antibody-super shift experiments. Taken together, the data presented in this paper provide evidence supporting the hypothesis that a stem-loop array of the 5'-upstream region of the human HLA-DRA gene could be directed by nuclear factors. In this manner, additional nuclear factors bound to the far region could be driven in close proximity of the transcription initiation site. PMID- 10385004 TI - Acyl-CoA synthetase catalyzes the synthesis of diadenosine hexaphosphate (Ap6A). AB - The synthesis of diadenosine hexaphosphate (Ap6A), a potent vasoconstrictor, is catalyzed by acyl-CoA synthetase from Pseudomonas fragi. In a first step AMP is transferred from ATP to tetrapolyphosphate (P4) originating adenosine pentaphosphate (p5A) which, subsequently, is the acceptor of another AMP moiety from ATP generating diadenosine hexaphosphate (Ap6A). Diadenosine pentaphosphate (Ap5A) and diadenosine tetraphosphate (Ap4A) were also synthesized in the course of the reaction. In view of the variety of biological effects described for these compounds the potential capacity of synthesis of diadenosine polyphosphates by the mammalian acyl-CoA synthetases may be relevant. PMID- 10385005 TI - Disordered C-terminal domain of tyrosyl-tRNA synthetase: secondary structure prediction. AB - The C-terminal domain (residues 320-419) of tyrosyl-tRNA synthetase (TyrRS) from Bacillus stearothermophilus is disordered in the crystal structure and involved in the binding of the anticodon arm of tRNA(Tyr). The sequences of 11 TyrRSs of prokaryotic or mitochondrial origins were aligned and the alignment showed the existence of conserved residues in the sequences of the C-terminal domains. A consensus could be deduced from the application of five programs of secondary structure prediction to the 11 sequences of the query set. These results suggested that the sequences of the C-terminal domains determined a precise and conserved secondary structure. They predicted that the C-terminal domain would have a mixed fold (alpha/beta or alpha+beta), with the alpha-helices in the first half of the sequence and the beta-strands mainly in its second half. Several programs of fold recognition from sequence alone, by threading onto known structures, were applied but none of them identified a type of fold that would be common to the different sequences of the query set. Therefore, the fold of the C terminal, anticodon binding domain might be novel. PMID- 10385006 TI - Pyrophosphate increases the efficiency of enterobactin-dependent iron uptake in Escherichia coli. AB - Exogenous inorganic pyrophosphate increases the biomass yield of Escherichia coli. In this report, we show that the effect of pyrophosphate is related to iron uptake. We have found that addition of pyrophosphate, ammonium iron (III) citrate or iron (III) chloride, in M63 minimal medium containing 1.7 microM of iron, causes an increase in growth yield. In contrast to iron chloride or ammonium iron (III) citrate, exogenous pyrophosphate is deleterious to strains unable to synthesize enterobactin. Thus the positive effect of pyrophosphate is related to the enterobactin uptake system expressed in a low iron content medium. Pyrophosphate in minimal medium has a repressing effect on the expression of Fur regulated genes. In iron rich medium where enterobactin synthesis is strongly decreased, addition of pyrophosphate increases expression of Fur-regulated genes. Furthermore, this latter regulatory effect of pyrophosphate in iron-rich medium is enhanced in the absence of enterobactin synthesis. It has also been shown that addition of pyrophosphate protects the cell against the oxidative stress caused by the presence of hydrogen peroxide in an iron-rich containing medium. These results indicate that pyrophosphate acts as an iron-chelating agent, could trigger the enterobactin-dependent iron uptake system and could promote an increased binding of iron to enterobactin. PMID- 10385007 TI - Binding and internalization of p1,p4-diadenosine 5'-tetraphosphate by bovine aortic endothelial cells. AB - p1,p4-Diadenosine 5'-tetraphosphate (Ap4A) has been implicated as a modulator of blood vessel tone. We have recently demonstrated that the infusion of Ap4A into swine induces vasodilation (Hilderman et al., Am. J. Hypertension 10 (1997) 94A) and that Ap4A induces the release of nitric oxide (NO) from bovine aortic endothelial cells (BAEC) (Hilderman and Christensen, FEBS Lett. 427 (1998) 320 324). However, the interaction of Ap4A with endothelial cells is incompletely understood. Therefore, we determined the characteristics of [3H]-Ap4A binding to BAEC in normal and ATP-depleted cells. These binding studies demonstrate that the interaction of Ap4A with BAEC involves two distinct steps: an ATP independent step and a second ATP dependent step leading to internalization of Ap4A. The initial interaction of Ap4A with BAEC is not affected by either EGTA or iodoacetate; however, both agents block the second step. These data suggest that calcium ions and sulfhydryl groups are required for Ap4A internalization but not for an initial binding event. PMID- 10385008 TI - The amino acidic substitution of cysteine 167 by serine (C167S) in BstVI restriction endonuclease of Bacillus stearothermophilus V affects its conformation and thermostability. AB - The restriction endonuclease BstVI from Bacillus stearothermophilus V contains three cysteine residues at positions 134, 167 and 180. Titration of Cys residues with DTNB showed that none of them are involved in disulphide bond formation. Cysteine triplets 134 and 167 were modified by recombinant PCR to introduce a serine residue in each case. The mutated genes were cloned into pGEM-T vector and transformed into E. coli JM109. Even though pGEM-T is not designed for expression, the mutant proteins were efficiently expressed in E. coli. The endonuclease carrying the mutation C134S was purified to homogeneity but appeared to be very unstable. In contrast, the C167S mutant enzyme was stable when pure and was studied biochemically. This mutant enzyme was as stable and resistant to protein-denaturing agents as the wild type enzyme. The activity of both enzymes was not affected by preincubations of 2 h at 80 degrees C. A short preincubation at 95 degrees C caused a complete inactivation of the mutant enzyme while the wild type endonuclease retained 30% of its activity. Moreover, the C167S BstVI was more susceptible to be hydrolyzed by proteinase K and trypsine compared to the wild type endonuclease. These results show that the substitution Cys --> Ser at position 167 affects the configuration and thermostability of BstVI restriction endonuclease. PMID- 10385009 TI - Characterization of Aspergillus niger phosphoglucose isomerase. Use for quantitative determination of erythrose 4-phosphate. AB - Phosphoglucose isomerase (PGI) was purified from Aspergillus niger and the in vitro kinetic properties of the enzyme were related to its functioning in vivo. A new assay method was developed to study the forward reaction making use of mannitol 1-P dehydrogenase as the coupling enzyme. In this simple assay system mannitol 1-P dehydrogenase converts fructose 6-P and NADH to mannitol 1-P and NAD+, respectively. At pH 7.5 the Km for glucose 6-P was 0.48 mM, whereas the Km for fructose 6-P was 0.32 mM. The pentose phosphate pathway intermediates 6 phosphogluconate and erythrose 4-P (E4P) were competitive inhibitors of PGI with Ki values of approximately 0.2 mM and 1 microM respectively. In citric acid producing A. niger mycelium inhibition by 6-phosphogluconate is of minor physiological significance (10% inhibition). Since E4P could not be detected by an existing procedure, a novel assay was developed based on the strong inhibition of PGI by E4P. Although the new assay is very sensitive (detection limit 25 pmol), E4P could still not be detected in metabolite extracts indicating that a very low level of E4P is present in the cells. Using in vitro kinetics and concentrations of intracellular metabolites the in vivo activity of PGI was calculated and closely matched the steady state glycolytic flux observed during citric acid production. PMID- 10385010 TI - Prospective clinical and microbiological study of pleural effusions. AB - A prospective clinical microbiological study of pleural fluid samples was conducted to investigate the etiology of pleural effusions and to evaluate two different methods for transport and culture of these samples. A total of 245 pleural fluid specimens were inoculated into a transport vial, an aerobic and an anaerobic blood culture vial, and a sterile tube. One hundred nine samples were from infectious patients and 128 from noninfectious patients. Gram stain had a sensitivity of 48% and a specificity of 100% as compared to culture. Of the total, 15.5% of the samples were positive for microorganisms, and 60% of the positive samples were nonpurulent pleural fluid. Single-organism growth was found in 23 samples (60.5%). Sixty-three microorganisms were isolated: 25 (39.7%) aerobic, 22 (35%) anaerobic, 13 (20.6%) mycobacteria, and three (4.7%) fungi. Of the 25 positive samples, excluding those samples that grew mycobacteria, nine (36%) were positive exclusively in the blood culture vials. Twelve organisms were isolated, only one of which did not grow in the anaerobic vial. Two (8%) samples were positive by conventional culture only, and 14 (56%) were positive by both methods. The microorganism isolation rate obtained with use of blood culture vials was significantly greater than that obtained with the conventional method of transport and culture. Sixty-three percent of the empyema patients had an associated underlying pathology, pneumonia being the most frequent. In conclusion, for microbiological study of pleural fluid, it seems appropriate to inoculate all samples, including nonpurulent samples, into both a sterile tube and an anaerobic blood culture vial. PMID- 10385011 TI - An epidemic of pertussis among elderly people in a religious institution in The Netherlands. AB - An epidemic of pertussis is described among elderly people in a religious institution in the Netherlands in 1992. Subjects were evaluated for their vaccination status and for history and presence of respiratory symptoms. Specimens were collected for culture, polymerase chain reaction, and serological evaluation. None of the 75 residents and 19 of 24 nonresident personnel had been vaccinated against pertussis. The overall attack rate of clinical pertussis, defined as persistent cough lasting at least 2 weeks, was 49%. In five subjects with clinical pertussis, either culture or polymerase chain reaction or both were positive for Bordetella pertussis. A significant (at least 4-fold) change in specific antibody titre was observed in 85% (41/48) and 20% (10/49) of subjects with and without clinical pertussis, respectively (P < 0.0001, chi-square 41.1). The attack rate of laboratory-confirmed pertussis was 42% (41/98). This rate was 5% (1/19), 20% (1/5), and 53% (39/74) in vaccinated personnel, nonvaccinated personnel, and nonvaccinated residents, respectively (not significant). Among residents aged between 55-74 years and 75-94 years, the attack rates were 47% (17/36) and 58% (22/38), respectively (relative risk=0.8; 95% confidence interval 0.5-1.3). Four of 75 residents (5%) died from intracranial bleeding, while they were symptomatic for pertussis. It is concluded that the attack rate of pertussis was high among nonvaccinated elderly and that pertussis tended to increase with age. There may be a considerable risk of mortality from pertussis in this population. Physicians should be alert to the diagnosis of pertussis in the elderly with nocturnal and prolonged periods of coughing. PMID- 10385012 TI - Controlled clinical comparison of three commercial blood culture systems. AB - In a controlled clinical comparison, three commercial blood culture systems--the standard aerobic BacT/Alert bottle (STD), the aerobic BacT/Alert FAN bottle (FAN) and the Isolator system (ISO; Wampole Laboratories, USA) were compared for their ability to detect aerobic and facultatively anaerobic microorganisms. A total of 945 BacT/Alert (STD and FAN) blood culture sets were compared. Of these, 110 blood culture sets (11.6%) yielded growth of 116 clinically significant bacterial and fungal isolates. Microorganisms were recovered from 10.7% (101/945) of the FAN bottles compared to 8.9% (84/945) of the STD bottles. Of the significant isolates, 78 (67.2%) were recovered by both bottles, 29 (25%) by the FAN bottle only and nine (7.8%) by the STD bottle only (P<0.01). Along with 56.1% (530/945) of BacT/Alert blood culture sets, a concomitant ISO tube was obtained. Of the triple (STD + FAN + ISO) blood culture sets, 54 (10.2%) yielded growth of 59 clinically relevant isolates. Microorganisms were detected in 9.1% (48/530) of the FAN bottles, 8.3% (44/530) of the STD bottles and 4% (21/530) of the ISO tubes (P<0.001). Overall, the BacT/Alert system detected more clinically significant microorganisms than the ISO tube; the STD and the FAN bottle each recovered significantly more staphylococci (P<0.01 and P<0.001, respectively) and gram-negative rods (P<0.01, both). In conclusion, the BacT/Alert FAN bottle performed better than the BacT/Alert STD bottle; both BacT/Alert bottles, however, were superior to the ISO tube in terms of recovery of clinically significant microorganisms, including gram-positive and gram-negative bacteria. PMID- 10385013 TI - Comparison of three different commercial methods for measuring plasma viraemia in patients infected with non-B HIV-1 subtypes. AB - In order to determine whether commercial assays presently in use for the quantification of plasma human immunodeficiency virus type 1 (HIV-1) RNA levels detect different genetic viral subtypes with the same reliability, a panel of 38 samples corresponding to 22 HIV-1-infected patients, representing non-B subtypes A-F, was examined. One to three plasma samples belonging to each individual were tested by the second-generation HIV-1 branched DNA (bDNA) assay (Chiron, Spain), the Nuclisens assay (Organon-Teknika, Spain), the Amplicor Monitor reverse transcriptase polymerase chain reaction assay (Roche, Spain), and the Ultradirect Monitor (Roche) using primers specifically designed to amplify non-B HIV-1 subtypes. Each of the different methods for measuring viral load showed a distinct sensitivity for non-B HIV-1 subtypes. Values higher than the assay detection limit were obtained in 22 (57.9%), 33 (86.8%), and 37 (93.3%) samples using the bDNA, Nuclisens, and Monitor assays, respectively. Significantly different values (>0.5 logs) were found in 55.3%, 81.6%, and 71.1% of specimens comparing results provided by bDNA and Nuclisens, bDNA and Monitor, and Nuclisens and Monitor, respectively. Quantitative values provided by the Ultradirect Monitor test using non-B primers were particularly discordant with the other tests. Overall, 44.7% of samples yielded higher viral load values with this assay than with the regular Monitor assay, reflecting its enhanced sensitivity for non B subtypes; however, the reproducibility of this test was low. These results support the recommendation of always using the same assay when monitoring plasma viraemia. PMID- 10385014 TI - Molecular typing of erythromycin-resistant streptococcus pyogenes strains with the M phenotype isolated in Italy. AB - To assess the spread of the new M phenotype, various erythromycin-resistant Streptococcus pyogenes strains from three Italian cities (Verona, Monza, Florence) were characterised. Each strain was analysed for the presence of genes ermAM and mefA, for the ability to accumulate radioactive erythromycin in the absence of sodium arsenate, for the protein T serological type, and for the DNA macrorestriction profile identified by means of pulsed-field gel electrophoresis. In a number of strains, the presence of the inducible ermAM gene was demonstrated; all these strains were negative in the efflux-pump detection assay, did not possess the mefA gene, and had similar restriction profiles. The strains with the efflux mechanism and mefA gene belonged to different serotypes. Of these, only one serotype, T4, was isolated in all three cities. The restriction profile analysis with SmaI and SfiI revealed a very close correlation between strains with the same serotype. PMID- 10385015 TI - Fully automated liquid culture system compared with Lowenstein-Jensen solid medium for rapid recovery of mycobacteria from clinical samples. AB - The aim of this study was to compare the rate of recovery of mycobacteria and the time to detection in 5208 samples using the MB/BacT culture system (Organon Teknika, USA) and Lowenstein-Jensen medium. Mycobacteria were recovered from 301 (5.7%) samples. Two hundred fifty-seven (85.3%) isolates from 114 patients were Mycobacterium tuberculosis [135 (52.5%) smear-positive, 122 (47.4%) smear negative], and 44 (14.6%) were potentially pathogenic environmental mycobacteria. The yield with the MB/BacT was higher than that with Lowenstein-Jensen [287 (95.3%) vs. 200 (66.4%), P<0.001] for both Mycobacterium tuberculosis [247 (96.1%) vs. 187 (72.7%), P<0.001] and potentially pathogenic environmental mycobacteria [40 (90.9%) vs. 13 (29.5%), P<0.001], mainly at the expense of the smear-negative samples. Moreover, 70 (27.2%) samples were positive only in the MB/BacT, whereas ten (3.8%) samples were positive only in Lowenstein-Jensen. The number of patients with tuberculosis detected by the MB/BacT was higher than that detected by Lowenstein-Jensen medium [111 (97.3%) vs. 89 (78%), P<0.001]. In 25 (21.9%) patients the diagnosis was established solely by means of the MB/BacT. In smear-positive and smear-negative samples, the mean times to detection of Mycobacterium tuberculosis were 16.7 and 26.3 days, respectively, with Lowenstein Jensen and 11.5 and 19.3 days, respectively, with the MB/BacT. These results indicate that the MB/BacT is more efficient and faster than Lowenstein-Jensen for the recovery of mycobacteria. PMID- 10385016 TI - Genotypic resistance to zidovudine as a predictor of failure of subsequent therapy with human immunodeficiency virus type-1 nucleoside reverse-transcriptase inhibitors. AB - To define factors predictive of failure to respond to nucleoside reverse transcriptase inhibitors in human immunodeficiency virus type-1 (HIV-1)-infected subjects pretreated with zidovudine (ZDV), three groups of subjects shifted to double therapy with ZDV plus didanosine (ddI, n = 13), zalcitabine (ddC, n = 14), or lamivudine (3TC, n = 12) were retrospectively evaluated, with respect to addition of the second NRTI, at week 0 and week 24. Factors considered included duration of ZDV pretreatment, CD4+ cell counts, plasma HIV-1 RNA load, peripheral blood mononuclear cell HIV-1 DNA load, and HIV-1 DNA genotypic resistance to nucleoside reverse-transcriptase inhibitors. The three groups were well matched for baseline characteristics and did not differ significantly in virological and immunological response to the different combination treatments. Drug-specific resistance mutations were selected in more than half the cases by 3TC, but not by ddI and ddC. Low-level and substantial genotypic resistance to ZDV was detected 13 (33.3%) and in 19 (48.7%) patients at baseline, respectively, and evolved through week 24 in several patients. When subjects were divided into responders and nonresponders to the second nucleoside reverse-transcriptase inhibitor on the basis of a decrease of more than 0.5 log10 (n = 15) or less than 0.5 log10 (n = 21) in HIV-1 RNA, respectively, baseline genotypic ZDV resistance was the only independent predictor of failure in a logistic regression model (P = 0.003 or P = 0.024, depending on whether low-level resistance was considered or not, respectively). Thus, selection of ZDV resistance mutations may impair subsequent use of different nucleoside reverse-transcriptase inhibitor compounds. PMID- 10385017 TI - Procalcitonin concentrations in patients with neutropenic fever. AB - To assess the usefulness of markers of inflammation in distinguishing bacterial infection from severe systemic nonbacterial inflammation, concentrations of procalcitonin, neopterin, endotoxin, tumor necrosis factor, and interleukin-6 were measured in 28 neutropenic patients at the onset of fever and twice thereafter at 4 h intervals. Infection was found in 11 patients, and 17 patients had fever of undetermined origin. The procalcitonin concentration increased rapidly in patients with infection: the response was detectable within 8 h of the onset of fever. Procalcitonin is a specific but not a sensitive marker of infection in patients with neutropenic fever. Its poor sensitivity was related to an absent or delayed response in patients with gram-positive infections. Considerable overlap between infected and noninfected patients was found in levels of endotoxin, tumor necrosis factor, and interleukin-6. PMID- 10385018 TI - Outbreak of Escherichia coli O157 infection associated with a music festival. AB - Seven persons who attended the Glastonbury Music Festival were infected with Vero cytotoxin-producing Escherichia coli O157 and an eighth person had serological evidence of infection. Cases were reported from different parts of England. Patients were interviewed by telephone about clinical symptoms, festival attendance, camping details, food history, water exposure, and contact with mud and animals. The interviews identified no common food source, differing use of water sources and widely dispersed camping sites. Escherichia coli O157 strains from seven persons and from a cow belonging to a herd that had previously grazed the site all belonged to phage type 2 and possessed genes for Vero cytotoxin 2. Drug resistance and DNA-based tests showed that six patients were infected with strains indistinguishable from each other and from the bovine isolate. The most likely vehicle of infection was mud contaminated with Escherichia coli O157 from infected cattle. PMID- 10385019 TI - Staphylococcus lugdunensis endocarditis in a young, previously healthy female. AB - Reported here is a case of native valve infective endocarditis in which Staphylococcus lugdunensis was isolated from blood cultures as well as from the mitral valve. It is suggested that tube coagulase-negative staphylococci isolated from blood cultures of patients with infective endocarditis be considered carefully for possible clinical significance. The use of the ornithine carboxylase test as a simple screening method for identification of Staphylococcus lugdunensis is recommended. PMID- 10385020 TI - Evolution of resistance among clinical isolates of Acinetobacter over a 6-year period. AB - The aim of this report was to study the evolution of susceptibilities of 1532 clinical isolates of Acinetobacter recovered over a period of 6 years. The minimal inhibitory concentrations (MICs) of 15 antimicrobial agents were determined for all the isolates. The respective percentages of resistant strains in the years 1991 and 1996 were as follows: ciprofloxacin, 54.4% and 90.4%; tobramycin, 33% and 71.8%; amikacin, 21% and 83.7%; ampicillin plus sulbactam, 65.7% and 84.1%; ceftazidime, 57.4% and 86.8%; ticarcillin, 70% and 89.4%; trimethoprim plus sulfamethoxazole, 41.1% and 88.9%; and imipenem, 1.3% and 80%. The MIC90s of ciprofloxacin, sparfloxacin, biapenem, meropenem, imipenem, cefepime, cefpirome, and rifampicin against 250 imipenem-resistant Acinetobacter strains were >32, >32, 128, >256, 256, >256, 256, and 16 mg/l, respectively. With serious infections, it was necessary to resort to the use of colistin, the only antibiotic active in vitro. PMID- 10385021 TI - Short-term treatment of pertussis with azithromycin in infants and young children. AB - A prospective, open, noncomparative study was conducted to assess the efficacy and safety of azithromycin given once daily for 3 or 5 days to eradicate Bordetella pertussis from the upper respiratory tract of infants and young children. Seventeen children received azithromycin in a dose of 10 mg/kg on day 1 followed by 5 mg/kg once daily for four consecutive days, and 20 were given 10 mg/kg once daily for 3 days. Seven days after the initiation of therapy, 33 of 35 (94.3%) patients had negative cultures for Bordetella pertussis. On day 14, cultures from all 34 evaluable patients were negative. These findings suggest that a controlled, comparative study of erythromycin versus short-term administration of azithromycin is justified. PMID- 10385022 TI - Association between primary cytomegalovirus infection and severe hemolytic anemia in an immunocompetent adult. AB - Hemolysis is a rare complication of cytomegalovirus infection in the immunocompetent adult, and the mechanisms responsible for it remain obscure. Guidelines for treatment have yet to be established, and the effectiveness of antiviral therapy has not been proven. In this report, an unusual case of primary cytomegalovirus infection manifested by severe hemolysis in an immunocompetent adult is presented. Treatment with ganciclovir (5 mg/kg b.i.d.) for 10 days and prednisolone (2 mg/kg/day) for more than 3 months suggests that both virological and immunological mechanisms are probably responsible for the hemolysis. PMID- 10385023 TI - In vitro activity of the new echinocandin antifungal, MK-0991, against common and uncommon clinical isolates of Candida species. AB - A broth macrodilution method, performed as recommended by the National Committee for Clinical Laboratory Standards, was used for comparative testing of the new echinocandin antifungal agent MK-0991 and fluconazole against 50 yeast isolates belonging to 12 species of Candida. MK-0991 was shown to be highly effective against both fluconazole-susceptible and -resistant Candida spp., yielding minimum inhibitory concentrations ranging from < or = 0.19 to 0.78 microg/ml. Fungicidal activity was exerted at < or = 1.5 microg/ml for 73% of the isolates tested. This study suggests that MK-0991 has significant potential for clinical development. PMID- 10385024 TI - Antibacterial activity of moxifloxacin (Bay 12-8039) against aerobic clinical isolates, and provisional criteria for disk susceptibility tests. AB - Moxifloxacin (Bay 12-8039), ciprofloxacin, and levofloxacin were compared in vitro against 1074 clinical isolates gathered from different medical centers throughout North America during the winter months of 1997. Moxifloxacin E tests and broth microdilution tests gave comparable results. Moxifloxacin was particularly potent against respiratory pathogens such as Haemophilus influenzae and Streptococcus pneumoniae. Ciprofloxacin was the most potent study drug against the family of Enterobacteriaceae and Pseudomonas spp. For tests of 5 microg moxifloxacin disks, zone size criteria of < or = 17 mm for resistant (MIC > or = 8 microg/ml) and > or = 21 mm for susceptible (MIC < or = 2 microg/ml) are provisionally proposed for use while clinical trials are under way. PMID- 10385025 TI - Imported dengue virus type 2 infection acquired during an outbreak in India. PMID- 10385026 TI - Resistance of Salmonella and Campylobacter species to antimicrobial agents. PMID- 10385027 TI - Models for oxygen sensing in yeast: implications for oxygen-regulated gene expression in higher eucaryotes. AB - Adaptation to changes in oxygen tension in cells, tissues, and organisms depends on changes in the level of expression of a large and diverse set of proteins. It is likely that most cells and tissues possess an oxygen sensing apparatus and signal transduction pathways for regulating expression of oxygen-responsive genes. Although progress has been made in understanding the transcriptional machinery involved in oxygen-regulated gene expression of eucaryotic genes the underlying mechanism(s) of oxygen sensing and the signaling pathways that connect oxygen sensor(s) to the transcription machinery of eucaryotes are still poorly understood. The yeast Saccharomyces cerevisiae is ideal for addressing these problems. Indeed, it is well-suited for broadly based studies on oxygen sensing at the cellular level because it lends itself well to genetic and biochemical studies and because its genome has been completely sequenced. This review focuses on oxygen-regulated gene expression and current models for oxygen sensing in this yeast and then considers their applicability for understanding oxygen sensing in mammals. PMID- 10385028 TI - Carotid body glomus cells: chemical secretion and transmission (modulation?) across cell-nerve ending junctions. AB - Glomus cells of the carotid body contain and secrete chemicals during 'natural' stimulation (hypoxia, hypercapnia and acidity), thus, the birth of the 'transmitter hypothesis of chemoreception'. Released chemicals would cross the synaptic cleft between glomus cells and carotid nerve terminals to depolarize the nerve ending membrane during excitation and hyperpolarize the membrane during inhibition. The main problem with this hypothesis is that specific synaptic blockers modify but do not block the effects of natural stimulation, while blocking the effects of the putative transmitters. It is proposed in this review that the secretion of chemicals is modulated by changes in electric coupling between glomus cells and that glomus cell-nerve ending transmission is not blocked by specific blockers for two reasons. One is that multiple transmitters are released. The other, and more the likely explanation, is that there are electric connections between these elements allowing the flow of currents that are unaffected by the blockers. PMID- 10385029 TI - K+ currents of glomus cells and chemosensory functions of carotid body. AB - The mechanism by which the carotid body senses hypoxia is not resolved, but the glomus cell, a secretory cell apposed to the afferent nerve endings, is believed to play an essential role. It is proposed that hypoxia causes glomus cell depolarization, leading to activation of voltage-gated calcium influx and enhanced secretion of an excitatory transmitter. The initial step, hypoxia induced depolarization, may be mediated by several candidate K+ channels which are sensitive to hypoxia, including: (1) a transient, voltage-dependent current; (2) a calcium and voltage dependent current; and (3) a non-voltage dependent, leak K+ current. If these channels represent the initial step in the hypoxia transduction cascade then it would be expected that K+ channel blocking agents would mimic the hypoxia response, leading to glomus cell secretion and increased nerve activity. This has been tested for the first two channels which are sensitive to classical K+ channel blocking agents, and, in general, results have not borne out this prediction. At present, the pharmacology of the leak K+ channel is not determined, and the experiment has not been undertaken. Thus, at present, hypoxic inhibition to a K+ channel in the glomus cell may initiate chemotransduction but there are many unanswered questions, especially the failure of K+ channel blocking agents to emulate the hypoxic response. PMID- 10385030 TI - NO and CO as second messengers in oxygen sensing in the carotid body. AB - It is being increasingly appreciated that nitric oxide (NO) and carbon monoxide (CO) are synthesized in mammalian cells and that they function as second messengers. The purpose of this article is to highlight the current information on NO and CO in the carotid body and discuss their significance in oxygen chemoreception. The NO synthesizing enzyme, nitric oxide synthase, is localized to nerve fibers and vascular endothelium in the carotid body. In vitro biochemical assays have shown that acute hypoxia inhibits NO synthase activity in carotid body extracts. Prolonged hypoxia up-regulates mRNA's encoding neuronal and endothelial NO synthases in the carotid body. Physiological studies have shown that NO is inhibitory to the carotid body sensory activity and mediates efferent inhibition. The actions of NO are in part mediated by its effects on glomus cells, wherein NO modulates Ca2+ channel activity and affects [Ca2+]i. The carotid body also uses another highly related gas as a second messenger, carbon monoxide (CO). The enzyme responsible for CO biosynthesis, heme oxygenase-2, is localized to glomus cells. CO, like NO, also exerts an inhibitory influence on sensory activity. Some of the actions of CO are mediated by altering Ca2+ channel activity and [Ca2+]i in glomus cells. Molecular oxygen is essential for biosynthesis of NO and CO. Under normoxia, basal levels of NO and CO act as amplifiers of molecular oxygen and keep the sensory discharge low. During hypoxia, decreased synthesis of NO and CO may contribute in part to the augmentation of sensory discharge. PMID- 10385031 TI - Redox-dependent binding of CO to heme protein controls P(O2)-sensitive chemoreceptor discharge of the rat carotid body. AB - Simultaneous recordings of chemoreceptor discharge and redox state of cytochromes have been carried out on the rat carotid body in vitro under the influence of carbon monoxide (CO) in order to identify the primary oxygen sensor protein controlling transmitter release and electrical activity. CO excites in a photolabile manner chemoreceptor discharge under normoxic conditions and inhibits under hypoxic conditions probably by binding to heme proteins. We hypothesize that type I cells and adjacent nerve endings of the carotid body tissue have a different apparatus with oxygen sensing heme proteins to cooperate for the generation of peripheral chemoreceptor response. Transmitter release from type I cells might be established in a redox dependent manner whereas membrane potential of nerve endings might be controlled by a heme coupled to ion channels. PMID- 10385032 TI - Background leak K+-currents and oxygen sensing in carotid body type 1 cells. AB - One model of oxygen sensing by the carotid body is that hypoxia depolarises type 1 cells leading to voltage-gated calcium entry and the secretion of neurotransmitters which then excite afferent nerves. This paper revues the mechanisms responsible for the membrane depolarisation in response to hypoxia. It concludes that depolarisation is caused not through the inhibition of calcium activated or delayed rectifier K+-channels but through the inhibition of an entirely new type of background K+-channel. This channel lacks sensitivity to the classical K+-channel inhibitors TEA and 4-AP. New evidence does however reveal that background K+-channels in the type 1 cell can be inhibited by Ba2+ and that Ba2+ depolarises isolated type 1 cells. Intriguingly, Ba2+ is the only K+-channel inhibitor thus far reported to stimulate the carotid body. These studies therefore support the hypothesis that depolarisation of the type 1 cell is an integral part of the oxygen sensing pathway in the carotid body. PMID- 10385033 TI - Acetylcholine contributes to hypoxic chemotransmission in co-cultures of rat type 1 cells and petrosal neurons. AB - The neurotransmitter mechanisms that mediate chemosensory transmission in the mammalian carotid body (CB), i.e. the primary arterial P(O2) detector, are controversial. Given the inherent difficulty of recording from afferent terminals in situ, the authors have adopted an alternative approach based on co-culture of dissociated CB receptor (type 1) cell clusters and petrosal neurons (PN) from 8 14-day-old rat pups. Electrophysiological, perforated patch recordings from petrosal somas, juxtaposed to type 1 clusters, revealed the development of a high incidence of functional 'synapses' in vitro. Recent evidence has strengthened the case for acetylcholine (ACh) as a co-released transmitter: (i) cultured type 1 cells express several cholinergic markers including the vesicular ACh transporter (VAChT), intracellular acetylcholinesterase (AChE), and occasional clear cored vesicles (approximately 50 nm diameter); (ii) the frequency of spontaneous synaptic activity, as well as the hypoxia-induced depolarization recorded in 'juxtaposed' PN in co-culture, were partially suppressed by the nicotinic ACh receptor (nAChR) blocker, mecamylamine (2 microM); (iii) consistent with the presence of extracellular AChE, ACh-mediated membrane noise in type 1 cells as well as the hypoxia-evoked PN response in co-culture were potentiated in a few cases by the AChE inhibitor, eserine (100 microM). Thus, since many PN and type 1 cells express mecamylamine-sensitive nAChR, released ACh may act presynaptically on type 1 cell autoreceptors and/or postsynaptically on petrosal terminals. Other CB transmitter candidates (e.g. 5-HT and ATP) were found to excite PN, though their potential role as co-released sensory transmitters requires further investigation. PMID- 10385034 TI - Neuroepithelial bodies as airway oxygen sensors. AB - Since the discovery of neuroepithelial bodies (NEB) in the late 1930s, evidence has accumulated to suggest that these cells may function as hypoxia-sensitive airway sensors. Until recently, this hypothesis was based largely on morphological observations. The use of in vitro models of isolated NEB, combined with electrophysiological approaches, have provided direct evidence that NEB cells express a membrane-bound O2 sensor and are the transducers of hypoxic stimulus. Here, we review the historical evidence and current state of knowledge of the oxygen-sensing properties of NEB cells, comparison with other O2 sensing cells, as well as recent advances that have been made using molecular and electrophysiological techniques. The possible role of NEB in perinatal pulmonary pathophysiology is also discussed. PMID- 10385035 TI - K+ and Ca2+ channel activity and cytosolic [Ca2+] in oxygen-sensing tissues. AB - Ion channels are known to participate in the secretory or mechanical responses of chemoreceptor cells to changes in oxygen tension (P(O2)). We review here the modifications of K+ and Ca2+ channel activity and the resulting changes in cytosolic [Ca2+] induced by low P(O2) in glomus cells and arterial smooth muscle which are well known examples of O2-sensitive cells. Glomus cells of the carotid body behave as presynaptic-like elements where hypoxia produces a reduction of K+ conductance leading to enhanced membrane excitability, Ca2+ entry and release of dopamine and other neurotransmitters. In arterial myocytes, hypoxia can inhibit or potentiate Ca2+ channel activity, thus regulating cytosolic [Ca2+] and contraction. Ca2+ channel inhibition is observed in systemic myocytes and most conduit pulmonary myocytes, whereas potentiation is seen in a population of resistance pulmonary myocytes. The mechanism whereby O2 modulates ion channel activity could depend on either the direct allosteric modulation by O2-sensing molecules or redox modification by reactive chemical species. PMID- 10385036 TI - Roles for NAD(P)H oxidases and reactive oxygen species in vascular oxygen sensing mechanisms. AB - Observations that physiological levels of O2 control the rates of production of reactive O2 species by systems including NAD(P)H oxidases and that certain of these species have signalling mechanisms that regulate vascular tone has resulted in consideration of these systems in processes that mediate the sensing of changes in P(O2). Evidence exists for the participation of hydrogen peroxide dependent regulation of prostaglandin production and soluble guanylate cyclase activity, resulting from the metabolism of peroxide by cyclooxygenase and catalase, respectively, in P(O2)-elicited signalling mechanisms that regulate vascular force generation. A microsomal NADH oxidase whose activity is controlled by the redox status of cytosolic NAD(H) appears to function as a P(O2) sensor in bovine pulmonary and coronary arteries where changes in O2 levels control the production of superoxide anion-derived hydrogen peroxide and a cGMP-mediated relaxation response. Interactions with nitric oxide and superoxide anion, and the activity of glutathione peroxidase appear to influence the function of these O2 sensing systems, and some of these interactions, along with the activation of other oxidases, may contribute to alterations in P(O2) sensing mechanisms under pathophysiological conditions that affect vascular function. PMID- 10385037 TI - Oxygen sensing and signaling: impact on the regulation of physiologically important genes. AB - A growing number of physiologically relevant genes are regulated in response to changes in intracellular oxygen tension. It is likely that cells from a wide variety of tissues share a common mechanism of oxygen sensing and signal transduction leading to the activation of the transcription factor hypoxia inducible factor 1 (HIF-1). Besides hypoxia, transition metals (Co2+, Ni2+ and Mn2+) and iron chelation also promote activation of HIF-1. Induction of HIF-1 by hypoxia is blocked by the heme ligands carbon monoxide and nitric oxide. There is growing, albeit indirect, evidence that the oxygen sensor is a flavoheme protein and that the signal transduction pathway involves changes in the level of intracellular reactive oxygen intermediates. The activation of HIF-1 by hypoxia depends upon signaling-dependent rescue of its alpha-subunit from oxygen dependent degradation in the proteasome, allowing it to form a heterodimer with HIF-1beta (ARNT), which then translocates to the nucleus and impacts on the transcription of genes whose cis-acting elements contain cognate hypoxia response elements. PMID- 10385038 TI - Regulation of gene expression and secretory functions in oxygen-sensing pheochromocytoma cells. AB - The cellular response to hypoxia is complex. Specialized oxygen chemosensitive cells that are excitable respond to reduced O2 by membrane depolarization, altered gene expression, and neurotransmitter secretion. We have used the O2 sensitive pheochromocytoma (PC12) cell line to investigate the cellular response to hypoxia. Here, we present evidence that membrane depolarization and increased intracellular free Ca2+ are major regulatory events in these cells. Membrane depolarization is mediated by the inhibition of a slow-inactivating voltage dependent potassium (K) channel. Evidence from molecular biology and patch-clamp studies indicate that the O2-sensitive K channel is a member of the Kv1 family. We also reviewed findings on the regulation of gene expression in PC12 cells during hypoxia. An increase in intracellular free Ca2+ is required for hypoxia induced transcription of a number of genes including tyrosine hydroxylase (TH), the rate-limiting enzyme in the synthesis of catecholamine neurotransmitters, and several of the immediate early genes. We also reviewed the role of dopamine (DA) and adenosine (ADO) receptors in regulation of membrane depolarization and gene expression. PMID- 10385039 TI - Sleep, respiration and ALS. PMID- 10385040 TI - Counting muscular dystrophies in the post-molecular census. PMID- 10385041 TI - The effect of aniracetam on cerebral glucose metabolism in rats after lesioning of the basal forebrain measured by PET. AB - To evaluate the effect of aniracetam, a potent modulator of the glutamatergic and cholinergic systems, on the altered cerebral glucose metabolism after lesioning of the basal forebrain, we measured the cerebral metabolic rate of glucose (CMRGlc) with positron emission tomography and the choline acetyltransferase (ChAT) activity in the frontal cortex of the lesioned rats after treating them with aniracetam. Continuous administration of aniracetam for 7 days after the surgery prevented CMRGlc reduction in the frontal cortex ipsilateral to the lesion while the lesioned rats without aniracetam showed significant CMRGlc reduction in the frontal cortex. The level of CMRGlc in the lesion-side basal forebrain was lower in all rats regardless of the aniracetam treatment. Biochemical studies showed that aniracetam did not alter the reduction in the frontal ChAT activity. These results showed that aniracetam prevents glucose metabolic reduction in the cholinergically denervated frontal cortex with little effect on the cortical cholinergic system. The present study suggested that a neurotransmitter system other than the cholinergic system, e.g. the glutamatergic system, plays a central role in the cortical metabolic recovery after lesioning of the basal forebrain. PMID- 10385042 TI - Type IV phosphodiesterase inhibition in experimental allergic encephalomyelitis of Lewis rats: sequential gene expression analysis of cytokines, adhesion molecules and the inducible nitric oxide synthase. AB - Type IV phosphodiesterase inhibitors are able to suppress EAE. To investigate the effects of this therapy in the central nervous system, we serially analyzed from days 7 to 17 postinoculation the gene expression pattern of tumor necrosis factor (TNF), lymphotoxin, interferon-gamma, interleukin-1beta, the inducible nitric oxide synthase (iNOs), interleukin-10, the vascular cell adhesion molecule-1 (VCAM-1) and the intercellular adhesion molecule-1 (ICAM-1) in the spinal cord of Lewis rats with actively induced EAE, treated with Rolipram. Treated rats had a delayed and milder disease, and reduced numbers of infiltrates in the nervous tissue. The gene expression profile was similar to that of untreated rats, although delayed, with no evidence of IL-10 upregulation during the observation period. The delayed inflammation was not associated with changes in the expression of VCAM-1 and ICAM-1. In peripheral blood mononuclear cells, TNF mRNA levels were decreased and interleukin-10 was unchanged. This therapy did not alter the proliferative ability of T lymphocytes against myelin basic protein. The encephalitogenic potential of splenocytes from treated animals was also unaffected. The high levels of both iNOs mRNA and nitric oxide (NO) found before the appearance of clinical signs, suggests that NO generation might be a contributing factor to the therapeutic benefit achieved by Rolipram in the rat. PMID- 10385043 TI - Immunocytochemical studies of aquaporin 4 in the skeletal muscle of mdx mouse. AB - Immunostainability of anti aquaporin 4 antiserum was investigated in the muscles of dystrophin deficient mdx mice. Western blot analysis showed that the rabbit antiserum against aquaporin 4 reacted with a 28 kDa protein in extracts of normal mouse quadriceps femoris muscles but did not react with the protein in extracts of quadriceps femoris muscles of mdx mice. Immunoperoxidase staining of the muscles from normal and mdx mice revealed the positive immunoreaction at the myofiber surface of normal mice and the negative, or the faint and discontinuous immunostaining at the surface of mdx myofibers. Immunogold electron microscopy disclosed the localization of aquaporin 4 molecules at the myofiber plasma membranes of normal mice and the localization was consistent with that of orthogonal array particles in the protoplasmic face of normal muscle plasma membrane seen in freeze fracture replicas. This study demonstrated that the density of aquaporin 4 molecules was decreased in the muscle plasma membranes of mdx mice, resulting in the faulty function of mdx myofibers. PMID- 10385044 TI - Dynamic changes in glucose metabolism induced by thiamine deficiency and its replenishment as revealed by a positron autoradiography technique using rat living brain slices. AB - Dynamic changes in the cerebral glucose metabolic rate (CMRglc) before and after thiamine replenishment were investigated in living brain slices obtained from pyrithiamine-treated (PT) and pair-fed control rats by use of a positron autoradiography technique. Fresh rat brain slices (300 microm thick) were incubated with [18F]2-fluoro-2-deoxy-D-glucose ([18F]FDG) in oxygenated Krebs Ringer solution at 36 degrees C, during which serial two-dimensional images of [18F]FDG uptake in the slices were constructed on the imaging plates. The net influx constant (=K) of [18F]FDG was determined by a Patlak graphical method of the image data. Prior to thiamine pyrophosphate (TPP)-loading, the K value in the neurologically symptomatic PT was higher in all brain regions except the thalamus and mammillary body than the control, suggesting compensatory enhanced glycolysis. The rapid decrease in this heightened net influx constant immediately after TPP-loading was surmised to be due to activation of pyruvate oxidation with lactate as the substrate, with this inhibiting the glycolysis. From > or = 150 min after TPP-loading, the K value continued to show low values in the thalamus and mammillary body, which are regarded as the responsible sites for Korsakoff syndrome, whereas in all other sites recovery to control values was observed. These findings suggest that using this technique the quantitative evaluation of serial local changes in CMRglc from thiamine deficiency to after its replenishment may be useful in elucidating the pathophysiology and prognosis of Wernicke's encephalopathy. PMID- 10385045 TI - Sleep-disordered breathing at an early stage of amyotrophic lateral sclerosis. AB - Eighteen amyotrophic lateral sclerosis (ALS) patients having neither respiratory complaints nor subjective symptoms of sleep disturbance were studied by using an ambulatory multi-parameter monitoring system during sleep. They were divided into two groups: 11 patients with predominantly bulbar form and seven with non-bulbar form. After performing daytime pulmonary function tests, the presence of sleep disordered breathing (SDB) was evaluated by using a portable device. ALS patients did not show significant SDB as a whole, and the respiratory disturbance index (RDI) was not significantly different between the bulbar group and the non-bulbar group. However, three patients of the bulbar group showed significant SDB, and the patterns of apnea/hypopnea suggested that both bulbar weakness and minimal diaphragmatic weakness might cause SDB in ALS patients at an early clinical stage. Multi-parameter respiratory monitoring during sleep should be included in the routine evaluation of ALS patients at an early clinical stage, especially those with predominantly bulbar involvement, in order to predict early respiratory failure. PMID- 10385046 TI - Sarcoglycanopathies are responsible for 68% of severe autosomal recessive limb girdle muscular dystrophy in the Brazilian population. AB - Sarcoglycanopathies (SGPs) constitute a subgroup of limb-girdle muscular dystrophies (LGMD), where the primary defect in one sarcoglycan (SG) glycoprotein (alpha-SG, beta-SG, gamma-SG or delta-SG) results in a deficiency of the whole complex. Four genes, at 17q, 4q, 13q and 5q, encode the four glycoproteins, and mutations in these genes cause diseases called LGMD2D, 2E, 2C and 2F. To estimate the prevalence, relative proportions and clinical features of SGPs, we have studied the SG proteins in muscle biopsies of 140 patients (from 115 unrelated Brazilian families) with a clinical diagnosis of LGMD. Alpha-SG immunofluorescence analysis showed a positive staining pattern in 70% (80/115) of the families, a patchy pattern in 14% (16/115) and a negative pattern in 16% (19/115) of the families. All the 19 alpha-SG negative, and four of the 16 alpha SG patchy patients were also deficient for the other three SG proteins, confirming the diagnosis of SGP in 20% of the LGMD families. None of the positive alpha-SG patients were deficient for any of the other three SG proteins, supporting the view that the SG complex functions as a unit. DNA analysis for the four sarcoglycan genes showed that alpha-SG mutations accounted for 47%, beta-SG for 16%, gamma-SG for 16% and delta-SG for 21% of the cases. SG abnormalities were observed in only 8.5% of patients with milder LGMD forms, but were present in 68% of patients with a severe Duchenne-like course. The relatively high frequency of SGP among Brazilian people with LGMD may be due to the disproportionally high frequency of African Brazilian SGP patients with the same mutation (particularly among LGMD2C and 2F patients), suggesting a founder effect. Consanguinity is also common in our SGP families. PMID- 10385047 TI - Antibodies to GD3, GT3, and O-acetylated species in Guillain-Barre and Fisher's syndromes: their association with cranial nerve dysfunction. AB - We examined serum antibodies to the four fetal antigens GD3, O-acetyl GD3, GT3, and O-acetyl GT3 ganglioside in patients with Guillain-Barre syndrome (GBS) or its variant Fisher's syndrome (FS). The patients with FS more often had significant IgG antibodies against GD3, GT3, and O-acetyl GT3 than did the healthy controls. Furthermore, anti-GD3 and anti-GT3 IgG antibodies were more often significantly present in the patients with FS than in those with GBS. IgG antibody to GD3, GT3, and O-acetyl GT3 had a significant association with the presence of ophthalmoparesis. These antibodies, however, cross-reacted with GQ1b and we detected no antibodies which specifically reacted with fetal gangliosides. In addition, oculomotor involvement was more closely related to IgG antibodies to GQ1b than to those to fetal gangliosides. No evidence was obtained that the serum antibodies to these fetal gangliosides are associated with specific neurologic signs of cranial nerves. PMID- 10385048 TI - Intermittent rhythmic delta activity (IRDA) morphology cannot distinguish between focal and diffuse brain disturbances. AB - IRDA (intermittent rhythmic delta activity) is an abnormal generalized EEG pattern that is not specific to any single etiology and can occur with diffuse or focal cerebral disturbances. To determine whether different electrographic features of IRDA and associated EEG findings can differentiate underlying focal from diffuse brain disturbances, we performed a blind analysis of 58 consecutive EEGs with an IRDA pattern, recorded from 1993 until 1996, in which we evaluated posterior background activity, focal slowing and IRDA characteristics (frequency, distribution, duration, symmetry and abundance). The clinical diagnosis, state of consciousness and CT brain findings were retrieved from the patients' hospital records. There were 58 patients (33 females; mean age, 58+/-21 years). Twelve (21%) had only focal brain lesions, while 46 (79%) had diffuse brain abnormalities, (15 diffuse structural, 19 metabolic abnormalities, 12 postictal). Normal consciousness and focal EEG slowing were more frequent in patients with focal abnormalities, however, this was not statistically significant. Of the patients with focal abnormality, 11 (92%) had normal posterior background activity either bilaterally (n=4) or contralateral to the focal lesion (n=7). Bilaterally normal posterior background activity was observed in about 30% in both groups. Bilaterally abnormal posterior background activity was apparent in one patient (8%) with focal brain lesion and in 31 patients (67%) with diffuse brain abnormalities (P<0.0001). There were no significant differences in IRDA electrographic features between the focal group and the group with diffuse brain disturbances. We conclude that IRDA morphology cannot distinguish between focal and diffuse brain abnormalities. PMID- 10385049 TI - Excessive daytime sleepiness in myotonic dystrophy. AB - The aim of the present study was to assess whether or not there is any correlation between magnetic resonance imaging (MRI) abnormalities and excessive daytime sleepiness (EDS) in a consecutive series of patients with myotonic dystrophy (MD). The influences of nocturnal breathing abnormalities and sleep morphology on EDS were also evaluated. Ten MD patients were studied by means of an all-night polysomnographic recording, the multiple sleep latency test (MSLT) and MRI. Diagnosis of MD was established on the basis of the clinical and electrophysiological evidence of myotonia as well as of the characteristic genetic pattern. No patient had respiratory failure. Polysomnography and MSLT were also evaluated in ten healthy age-matched controls under the same environmental conditions. The mean MSLT value was significantly lower in patients than in controls. Five of the ten patients were found to have pathological EDS. The quantitative sleep variables and the nocturnal apnoeas in these five patients were not significantly different from those of the patients without EDS. As two patients did not undergo MRI because of claustrophobia, the MRI data were considered in eight patients. Corpus callosum (CC) atrophy was detected in four patients, whereas three patients showed hyperintense areas in the white matter. No correlation was found between EDS and MRI indexes of subcortical atrophy as well as volume of the hyperintense areas. By contrast, a correlation was found between the MSLT value and the reduction in the anterior area of the CC. Our data suggest that CC atrophy might occur in MD patients, and that the size of the CC anterior area might be associated with EDS. PMID- 10385051 TI - Rapid diagnosis of cryptococcal meningitis by microscopic examination of centrifuged cerebrospinal fluid sediment. AB - The classic India ink test is positive in only half of cryptococcal meningitis cases, and reliable, rapid cryptococcal antigen (CRAG) testing requires technical expertise and facilities not always available. We therefore examined cerebrospinal fluid (CSF) sediment using May-Giemsa, periodic acid-Schiff, and Gram stains in 16 patients with cryptococcal meningitis. The India ink test was positive in seven patients (44%), while microscopic examination of sediment revealed cryptococci in 13 (81%); in six of these 13 the India ink test was negative. Both methods failed to detect the pathogen in the remaining three patients. CRAG testing in CSF was negative in two patients (one with acquired immunodeficiency syndrome, one with diabetes mellitus) whose India ink test also was negative while cryptococci were identified in their CSF sediment. No false positives occurred with CSF May-Giemsa staining in 27 cases of aseptic meningitis with negative cultures for Cryptococcus. In all, microscopic examination of centrifuged and stained CSF sediment proved more sensitive for rapid diagnosis of cryptococcal meningitis than the India ink method, and in two of our patients cryptococci were seen in centrifuged CSF sediment despite negative CRAG and India ink tests. PMID- 10385050 TI - Clinical features and anti-neural reactivity in neuropathy associated with IgG monoclonal gammopathy of undetermined significance. AB - Neuropathy has been frequently reported in patients with IgG monoclonal gammopathy of undetermined significance (MGUS) but it is still unclear whether this association has clinical or pathogenetic relevance. In order to clarify the possible role of IgG MGUS in the neuropathy we correlated the clinical and electrophysiological features of the neuropathy with the duration and anti-neural activity of the M-protein in 17 patients with neuropathy and IgG MGUS. Ten patients (59%) had a chronic demyelinating neuropathy clinically indistinguishable from chronic inflammatory demyelinating polyneuropathy (CIDP) while 7 (41%) had a predominantly sensory axonal or mixed neuropathy. In 80% of patients in the CIDP-like and 28% in the sensory group the IgG M-protein became manifest several months to years after onset of the neuropathy. Antibodies to one or more neural antigens (including tubulin, a 35KD P0-like nerve myelin glycoprotein, GD1a, GM1 and chondrotin sulfate C) were found in 40% of patients with CIDP-like and 43% with sensory neuropathy but also in 37% patients with IgG MGUS without neuropathy. Neuropathy associated with IgG MGUS is probably less heterogeneous than previously considered suggesting that this association may not be merely casual. The evidence for primary pathogenetic role of IgG M-proteins in the neuropathy remains however elusive. PMID- 10385052 TI - Bcl-2 and Bax protein expression in human myopathies. AB - Expression of the apoptosis-related proteins Bcl-2 and Bax was analysed by means of immunohistochemistry in muscle biopsies from 13 patients suffering from different muscular diseases (inclusion body myositis n=4, polymyositis n=3, Becker muscular dystrophy n=4, necrotizing myopathy n=2, and controls n=4), in an attempt to learn about the role of these proteins in human muscle diseases associated with muscle fiber necrosis and regeneration. Increased Bcl-2 and Bax immunoreactivity was observed as fine granular precipitates in the cytoplasm or as subsarcolemmal aggregates in pathological cases. Increased Bcl-2 and Bax immunoreactivity was detected in some necrotic fibers, regenerating fibers, ring fibers and some apparently normal muscle fibers. In addition, increased Bcl-2 and Bax immunoreactivity was observed in the periphery of rimmed vacuoles and in muscle fibers with abnormal mitochondria in patients suffering from inclusion body myositis. Double-labelling immunohistochemistry disclosed co-localization of both proteins in about 50% of Bcl-2-immunoreactive fibers. Finally, double labelling immunohistochemistry using N-CAM antibodies revealed that some Bax positive fibers were regenerating fibers. Since increased Bax immunoreactivity was not restricted to necrotic muscle fibers, the present results suggest that over-expression of this protein in human myopathies is probably independent of the process of cell death. PMID- 10385054 TI - Cells from individuals with SOD-1 associated familial amyotrophic lateral sclerosis do not have an increased susceptibility to radiation-induced free radical production or DNA damage. AB - Oxidative stress may play a role in the pathogenesis of familial amyotrophic lateral sclerosis (FALS). Superoxide dismutases (SODs) are enzymes that can influence free radical processes in irradiated cells and there is some evidence that manipulation of SODs can affect survival of cells after radiation treatments. SOD-1 associated FALS mutants may have an altered radiation response due to an enhanced generation of hydroxyl radicals or a compromised ability to neutralize free radicals. We have investigated the ability of the lymphoblastoid cell lines from FALS patients with SOD-1 gene mutations, patients with sporadic ALS and controls to handle oxidative stress induced by ionising radiation by measuring levels of intracellular reactive oxygen species and production of DNA double-strand breaks. Levels of reactive oxygen species, expressed as the slope of the relative fluorescence of a radical-reactive fluorochrome, in the cells from familial ALS patients with SOD-1 gene mutations (2.14+/-1.06 Gy(-1)) and patients with sporadic ALS (1.38+/-0.21 Gy(-1)) were not significantly different from the controls (1.54+/-0.39 Gy(-1)). No significant difference was observed in the production of DNA double-strand breaks between three groups. The ability of lymphoblastoid cells from FALS patients with SOD-1 gene mutations to scavenge radiation-induced free radicals is not compromised nor is their ability to protect DNA damage induced by ionising radiation. PMID- 10385053 TI - Bipap improves survival and rate of pulmonary function decline in patients with ALS. AB - Amyotrophic Lateral Sclerosis (ALS) is a progressive motor neuron disease that frequently causes death within five years of diagnosis. The majority of deaths are due to pulmonary complications resulting from respiratory muscle weakness and bulbar involvement. A promising respiratory intervention is the recently introduced bi-level intermittent positive pressure (Bipap), which is a noninvasive ventilator modality shown to reduce the work of breathing and improve not only gas exchange, but also exercise tolerance and sleep quality. The aim of this study was to assess the utility of Bipap in prolonging survival in ALS. We retrospectively analyzed the results of Bipap use in 122 patients followed at Hahnemann University. All patients in this study were offered Bipap when their forced vital capacity (FVC) dropped below 50% of predicted value. Group 1 (n=38) accepted Bipap and used it more than 4 h/day. Group 2 (n=32) did not tolerate Bipap well and used it less than 4 h/day. Group 3 (n=52) refused to try Bipap. There was a statistically significant improvement in survival from initiation of Bipap in Group 1 (14.2 months) compared to Group 2 (7.0 months, P=0.002) or 3 (4.6 months, P<0.001) respectively. Furthermore, when the slope of vital capacity decline was examined, the group that used Bipap more than 4 h/day had slower decline in vital capacity (-3.5% change/month) compared to Group 2 (-5.9% change/month, P=0.02) and Group 3 (-8.3% change/month, P<0.001). We conclude that Bipap can significantly prolong survival and slow the decline of FVC in ALS. Our results suggest that all patients with ALS be offered Bipap when their FVC drops below 50%, at the onset of dyspnea, or when a rapid drop in %FVC is noted. PMID- 10385055 TI - Brain cancer incidence in the provinces of Zaragoza and Navarre (Spain): effect of age, period and birth cohort. AB - Several studies have detected increases in malignant brain tumour incidence and mortality rates particularly among the elderly. We analyzed time trends in malignant brain tumors incidence in Zaragoza over the period 1973-1990 and Navarre over the period 1973-1991, two Spanish provinces that have been collecting data through their respective Cancer Registries for the last 20 years, using Poisson regression analysis of age, period of diagnosis and cohort. In general, age-adjusted rates showed a steady rise in both registries, except in the case of females in Navarre, for whom a decrease in risk was observed for the last period, 1988-1991. This increase is a reflection of the rise in incidence experienced by the elderly, since the cohorts successively register rates that are stable over time, and even downward in the case of females in Navarre. The risk run by generations born circa 1920-1930 was the highest encountered. Rates were higher in Navarre in both sexes and for all but the last period in females, when rates on the two registers stood level. Increasingly generalised use of CT scanning and magnetic resonance in the 1980s in Spain, coupled with better and more effective health care access for the elderly, are factors that may well have some bearing on these findings. PMID- 10385056 TI - Conjugated bilirubin decreases the biliary excretion of phospholipids. PMID- 10385057 TI - Cholestasis of pregnancy. AB - Cholestasis of pregnancy is the commonest liver disease unique to pregnancy and is characterized by pruritus in the mother in late pregnancy, without any skin rashes. This is accompanied by an elevation of the serum bile acids. Liver function test abnormalities may occur. Abdominal pain is not a feature and liver failure does not occur. The diagnosis is made by a suggestive history and exclusion of other causes by the history, serology and an upper abdominal ultrasound. All symptoms and signs should disappear within 4 weeks post-partum; prolonged post-partum courses should prompt a search for other causes, such as primary biliary cirrhosis. The syndrome is associated with a five-fold increased incidence of stillbirth, intra-partum foetal distress and pre-term labour. The reason is not clear and not predictable. The accepted management is induction or delivery at 38 weeks, which has led to a reduction in poor foetal outcome. Preliminary studies using ursodeoxycholic acid show symptomatic and biochemical improvement in most women treated. There is also a suggestion of an improved foetal outcome and treatment should be considered in women who present with the condition earlier in pregnancy. PMID- 10385058 TI - Helicobacter pylori: evidence for spouse-to-spouse transmission. AB - BACKGROUND: Spouse-to-spouse transmission of Helicobacter pylori infection still remains controversial. METHODS: We studied the prevalence of H. pylori infection among spouses of H. pylori-positive or -negative individuals and looked for intraspousal transmission. Twenty-five couples were studied. Initially, one individual per couple was selected as the index subject. Spouses of these H. pylori-positive or -negative index individuals underwent screening for H. pylori by serology, the rapid urease test and histology. Those couples in whom only one spouse was positive were followed up and H. pylori status was re-evaluated using the above tests after approximately 1 year in the H. pylori-negative spouse. RESULTS: Of 25 randomly selected index subjects, 18 were H. pylori positive and seven were negative. There was no significant difference in age, sex, socioeconomic status, presence of dyspeptic symptoms, duration of marriage and number of children in these index subjects. Spouses of H. pylori-infected index subjects had a significantly higher prevalence of H. pylori infection than the spouses of H. pylori-negative index subjects (83.3 vs 28.5%; P < 0.01). Age, sex, socioeconomic status, dyspeptic symptoms, duration of marriage and number of children were not different in H. pylori-positive or -negative spouses of H. pylori-positive index subjects. There were five such couples in whom only one spouse was H. pylori positive initially. At follow up, three of these H. pylori negative spouses became positive. These findings suggest person-to-person transmission or common source exposure within couples. PMID- 10385059 TI - Role of anti-Helicobacter pylori treatment in H. pylori-positive and cytoprotective drugs in H. pylori-negative, non-ulcer dyspepsia: results of a randomized, double-blind, controlled trial in Asian Indians. AB - BACKGROUND: The efficacy of anti-Helicobacter pylori treatment and cytoprotective drugs in H. pylori-positive and -negative non-ulcer dyspepsia (NUD), respectively, is debatable. METHODS: In a randomized study, the efficacy of anti H. pylori treatment versus sucralphate was tested in patients with NUD. One hundred and twelve patients with NUD, 62 positive and 50 negative for H. pylori were studied. Of 62 patients positive for H. pylori, 32 were treated with triple therapy (colloidal bismuth subcitrate, tetracycline and metronidazole) for 2 weeks and the remaining 30 were treated with sucralphate (1 g, q.i.d.) for 4 weeks. Of 50 patients negative for H. pylori, 25 each were treated with either sucralphate (1 g, q.i.d.) or ranitidine (150 mg, b.d.) for 4 weeks. RESULTS: In patients with NUD and H. pylori infection, triple therapy eradicated H. pylori in 88% and was superior to sucralphate in producing symptom relief (81 vs 33%, P = 0.0003) and histological improvement in gastritis (73 vs 30%, P = 0.003). In the H. pylori-negative group, sucralphate was superior to ranitidine with regard to symptom relief (68 vs 36%, P = 0.04) and improvement in gastritis (44 vs 12%, P = 0.09). The symptomatic improvement persisted until 12 weeks after the start of treatment in triple therapy group only. CONCLUSIONS: In patients with NUD associated with H. pylori, triple therapy was better than sucralphate in terms of symptomatic and histological improvement. However, sucralphate was superior to ranitidine in providing symptom relief in patients with H. pylori-negative NUD. PMID- 10385060 TI - Effects of ammonia solution on the gastric mucosa in cirrhotic rats and therapeutic effects of geranylgeranylacetone. AB - BACKGROUND: We designed an animal model in order to clarify whether Helicobacter pylori infection causes the gastric mucosal lesion frequently seen in cirrhotic patients. METHODS: Ammonia (NH3) solution was given to rats with carbon tetrachloride-induced cirrhosis. The gastric mucosal hexosamine (Hx) content and histopathological findings in the cirrhotic rats were analysed and compared with those of the intact liver rats. Moreover, the usefulness of geranylgeranylacetone (GGA) was investigated in both rat groups. Both rat groups were subdivided according to the treatment as follows: a control group, an NH3 (0.02% solution) group, and an NH3 + GGA (400 mg/kg per day) group; and fed for 4 weeks. RESULTS: The gastric mucosal Hx content of the control group of the cirrhotic rats (16.7 +/- 5.2 microg/mg) was significantly lower than that of the control group of the intact liver rats (26.6 +/- 4.5 microg/mg, P < 0.05). In the cirrhotic rats, the Hx content of both the NH3 (31.9 +/- 13.1 microg/mg) and the NH3 + GGA group (31.9 +/- 9.8 microg/mg) was significantly higher than the Hx content of the control group (P < 0.05). Microscopically, in the cirrhotic rats, while scattered mucosal erosions were recognized in three of five rats of the NH3 group, there were no erosions in any rats of the NH3 + GGA group. CONCLUSIONS: These data suggest that the gastric mucosal defence mechanism is defective in liver cirrhosis and that NH3 enhances this defensive mechanism by acting as mild irritant; however, in some cirrhotics this results in gastric erosion due to excessive irritation. Geranylgeranylacetone protects the gastric mucosa against NH3 irritation in cirrhotics without enhancing the Hx content. Thus, H. pylori infection may be a possible cause of the gastric mucosal lesion in patients with liver cirrhosis. The mechanism of the therapeutic effect of GGA is not due to an enhancement of the gastric mucosal Hx content. PMID- 10385061 TI - Case report: A case of lymphoepithelioma-like carcinoma of the oesophagus and review of the literature. AB - A 78-year-old Japanese female was admitted to our hospital with dysphagia and weight loss. An oesophageal tumour was demonstrated radiologically and endoscopically, and was diagnosed as oesophageal cancer by biopsy. Histologically, the resected tumour showed poorly differentiated squamous cell carcinoma with prominent lymphoid stroma and was diagnosed as the so-called lymphoepithelioma-like carcinoma (LELC). Epstein-Barr virus in the tumour was negative by polymerase chain reaction and in situ hybridization. Oesophageal LELC is extremely rare. The cases in the literature, as well as the one reported here, presented with gross features of a submucosal tumour-like appearance. Although the differentiation of the tumour cells is often poor, prognosis seems to be better than for other types of oesophageal cancer. Oesophageal LELC has characteristic clinicopathological features and should be classified by criteria independent of other types of tumour. PMID- 10385062 TI - Evaluation of endoscopic ultrasonography as an indicator for surgical treatment of gastric cancer. AB - BACKGROUND AND METHODS: Clinicopathological analysis of 346 patients with gastric cancer was made retrospectively and new criteria for the indication of a limited operation using endoscopic ultrasonography (EUS) was developed. Suggested new criteria for selecting gastric cancer patients for the limited operation were: (i) the cancer is located in the mucosa and the lymph nodes are not involved as indicated by EUS examination; (ii) the maximum size of the tumour is less than 2.0 cm; (iii) there are no multiple gastric cancers or simultaneous abdominal cancers; and (iv) the mucosal cancer of elevated type less than 2.0 cm is excluded because there are good indications for endoscopic mucosal resection. RESULTS AND CONCLUSIONS: We applied these new criteria to 262 patients and found that the patients who had limited operation had the same prognosis and a better quality of life compared with patients who had the conventional operation. PMID- 10385063 TI - Calcitriol for bone disease in patients with cirrhosis of the liver. AB - BACKGROUND: Osteoporosis is associated with cirrhosis of the liver, but the effects of therapy for osteoporosis associated with cirrhosis are still controversial. METHODS: We evaluated the effects of calcitriol (1alpha,25 dihydroxyvitamin D3) on bone mineral density (BMD) in 76 patients (26 men and 50 women) with cirrhosis who were assigned randomly to receive calcitriol (0.5 mg twice per day) or not. The BMD of the lumbar vertebrae was measured by dual energy X-ray absorptiometry at least twice, 12-57 months apart. RESULTS: For men, the mean annual change in BMD was 1.1% in the treated group and -0.4% in the control group. The median (25th and 75th percentiles) annual change in BMD was 0.6 (-0.1, 2.1%) in the treated group and -1.4 (-1.9, 1.6%) in the control group. The difference in the median annual change between the two groups was significant (P = 0.013). For women, the mean annual change in BMD was -0.5% in the treated group and -2.3% in the control group. The median (25th and 75th percentiles) annual change in BMD was -0.5 (-1.8, 1.3%) in the treated group and -1.5 (-3.8, 0.7%) in the control group. This difference was significant (P = 0.011). CONCLUSIONS: Our results suggest that calcitriol can prevent bone loss and, therefore, may be useful for the treatment of bone disease in patients with cirrhosis of the liver. PMID- 10385064 TI - Case report: Hepatocellular carcinoma in type 1a glycogen storage disease with identification of a glucose-6-phosphatase gene mutation in one family. AB - A 40-year-old man with glycogen storage disease type 1a (von Gierke disease, GSD1a) developed hepatocellular carcinoma (HCC). Cold single-strand conformation polymorphism (SSCP) with 12% glycerol identified the G727T mutation in the glucose-6-phosphatase (G6Pase) gene, which has been reported to be the most common mutation in Japanese GSD1a patients. This case report is the first documentation of HCC in a case with G727T mutation. Given the prevalence of HCC in GSD1a with various germline mutations, analysis is needed to confirm that the germline mutation in this case is really related to hepatocarcinogenesis. DNA analysis of the family pedigree of this case, revealed three individuals with GSD1a and seven heterozygous carriers of the G727T mutation. As the diagnosis of GSD1a in this family was made only after these three patients reached adulthood, DNA diagnosis may help early identification of GSD1a patients and prevention of the progression of the disease. This DNA-based diagnosis permits prenatal diagnosis in at-risk patients and may facilitate screening and counselling of patients clinically suspected of having this disease. PMID- 10385065 TI - Limitations of imaging diagnosis for small hepatocellular carcinoma: comparison with histological findings. AB - BACKGROUND AND AIMS: The purpose of this study was to clarify the value and limitation of imaging modalities for diagnosing small hepatocellular carcinoma (HCC). METHODS: Nodules (n = 207) with diameters of 20 mm or less detected by periodic ultrasonography and computed tomography in 139 patients with chronic liver disease were investigated with digital subtraction angiography (DSA) and magnetic resonance imaging (MRI). These findings were compared with histological findings. RESULTS: Histological diagnoses were adenomatous hyperplasia (AH, n = 27), well-differentiated HCC (n = 99), moderately differentiated HCC (n = 79) and poorly differentiated HCC (n = 2). We compared two groups: group A (n = 62), nodules of 10 mm diameters or less; and group B (n = 145), nodules 11-20 mm. Adenomatous hyperplasia accounted for approximately 30% of group A, but was difficult to diagnose with imaging modalities alone. We diagnosed those nodules showing hypervascular staining on DSA or hyperintensity on MRI T2-weighted images as HCC. Imaging alone was sufficient to diagnose HCC in 58% of the well differentiated nodules and 87% of the moderately and poorly differentiated nodules (P < 0.01). It was possible to diagnose HCC by imaging alone in 60% of all nodules or 45% of group A and 68% of group B (A vs B, P < 0.005). CONCLUSIONS: With decreasing differentiation and increasing diameter of nodules, the use of imaging modalities to diagnose HCC improved. Tumour biopsy was required to diagnose 55% of the cases in group A and 32% of the cases in group B. PMID- 10385066 TI - Use of scintigraphy with 99mtechnetium galactosyl human serum albumin for staging of primary biliary cirrhosis and assessment of prognosis. AB - BACKGROUND: Conventional models for prediction of survival in patients with primary biliary cirrhosis (PBC) are based on the results of blood tests and on the clinical condition, which may be affected by treatment. We evaluated the usefulness of hepatic receptor imaging with [99mtechnetium] diethylenetriaminepentaacetic acid galactosyl human serum albumin (GSA) for the staging and prognosis of PBC without the need for reference to laboratory test results. METHODS: The subjects were 45 patients with PBC, 10 healthy subjects, 62 patients with chronic hepatitis and 144 patients with cirrhosis. Computer acquisition of gamma-camera data was started just before the injection of 185 MBq [99mTc]-GSA and was stopped 20 min later. Time-activity curves were generated from regions of interest (ROI) for the heart and liver. A receptor index was calculated by dividing the radioactivity of the liver ROI by that of the liver plus-heart ROI 15 min after the injection. An index of blood clearance was calculated by dividing the radioactivity of the heart ROI 15 min after the injection by that of the heart ROI 3 min after the injection. RESULTS: The median receptor index was higher in patients with PBC than in those with cirrhosis. Among patients with PBC, the receptor index was lower in those with stage IV disease than in those in stages I, II or III. The index of blood clearance was lower in patients with PBC than in those with cirrhosis. Among patients with PBC, the index of blood clearance was higher in those with stage IV disease than in those in stages I, II or III. The receptor index was correlated significantly both to the risk score of the Mayo model and to the prognostic index of the Japanese model. The index of blood clearance was also correlated significantly to this score and prognostic index. CONCLUSIONS: Hepatic receptor imaging with [99mTc]-GSA is useful for the evaluation of hepatic functional reserve, staging of PBC and assessment of prognosis. PMID- 10385067 TI - Hepatitis E infection in children: study of an outbreak. AB - BACKGROUND: Hepatitis E virus (HEV) is responsible for most of the hepatitis epidemics in the developing world and it frequently affects young adults. Therefore, common perception is that it does not affect children. METHODS: A group of 20 school children (13 years old) were possibly exposed to hepatitis E virus infection during a 2 day trekking trip. Epidemiological and clinical information was correlated to the presence of the hepatitis E virus genome and antibodies to HEV structural and non-structural proteins found in the blood of the children, using polymerase chain reaction and line immunoassay techniques. RESULTS: Ten children developed icteric hepatitis, seven prodrome-like illness without jaundice while three remained asymptomatic. Immunoglobulin M (IgM) antibodies to open reading frame (ORF)2 protein (pORF2) were detected in all 19 children tested, whereas 11 and 10 of the children were positive for IgM antibodies against ORF1 (pORF1) and ORF3 (pORF3) proteins, respectively. The rate of HEV infection was found to be 85%. Viraemia was observed in 11 children and was present in four of the seven anicteric patients (55%) compared with six of the nine (66%) icteric patients. One child without any symptom also had viraemia. CONCLUSIONS: The data obtained indicate a high susceptibility of children for HEV infection and a frequently prolonged viraemia in those infected. PMID- 10385068 TI - Effects of bilirubin ditaurate on biliary secretion of proteins and lipids: influence on the hepatic vesicle transport system. AB - BACKGROUND: Several organic anions cause dissociation of biliary lipid secretion from bile acid secretion (uncoupling). As bile lipids originate from liver microsomes and are transported by carrier proteins and/or transcytotic vesicles, such a reduction of biliary lipid secretion may lead to cytosolic accumulation of vesicles. This study investigated whether bilirubin conjugate, a physiologically important organic anion, caused uncoupling and whether hepatic retention of compounds carried by transcytotic vesicles occurred subsequently, using bilirubin ditaurate, a synthetic commercially available compound. METHODS: Cannulation of the bile duct and femoral vein was done in male Sprague-Dawley rats. Sodium taurocholate was infused intravenously at a constant rate of 100 nmol/min per 100 g bodyweight. Bilirubin ditaurate (50 nmol/min per 100 g bodyweight) was infused concomitantly, followed by periodical bile collection for analysis of lipids, total protein and immunoglobulin A. RESULTS: Biliary bile acid secretion was not changed significantly by infusion of bilirubin ditaurate. In contrast, the secretion of cholesterol, phospholipids and immunoglobulin A was decreased by 57.3, 48.7 and 44.8%, respectively. The biliary cholesterol:phospholipid ratio was increased by 19%. Uncoupling was caused by bilirubin ditaurate and biliary immunoglobulin A secretion was decreased. CONCLUSIONS: As immunoglobulin A is a major protein carried by intrahepatic transcytotic vesicles, uncoupling may involve impairment of intrahepatic vesicular transport. Also, a reduction of immunoglobulin A secretion into bile by organic anion-induced uncoupling may weaken biliary immunity. PMID- 10385069 TI - Effect of pancreastatin on cerulein-stimulated pancreatic blood flow and exocrine secretion in anaesthetized rats. AB - BACKGROUND: Pancreastatin (PST) is an inhibitor of pancreatic exocrine secretion in vivo but not in vitro, which suggests that the inhibitory effect of PST is indirect, that is, not mediated by a specific receptor on pancreatic acinar cells. In this study, we investigated the effects of PST on pancreatic exocrine secretion and local pancreatic blood flow in anaesthetized rats to elucidate the participation of PST in indirect regulation of pancreatic exocrine function through blood supply. METHODS: Pancreastatin (100, 200 or 500 pmol/kg per h) was administered intravenously under background infusion of cerulein (0.5 microg/kg per h), a cholecystokinin analogue. Pancreatic exocrine secretion was monitored by volume and protein output of the pancreatic juice and local pancreatic blood flow was measured by the hydrogen gas clearance method. RESULTS: Pancreastatin significantly reduced cerulein-induced local pancreatic blood flow in a dose dependent manner. Pancreatic exocrine secretion was also reduced significantly by PST dose-dependently. Pancreastatin did not change systemic blood pressure. These results suggested that the reduction of pancreatic blood flow is associated with the reduction of pancreatic exocrine secretion. CONCLUSIONS: We conclude that the mechanism of PST-induced inhibition of pancreatic exocrine secretion is, at least, partly mediated by the reduction of local pancreatic blood flow through blockade, caused by the action of cerulein on pancreatic blood flow. PMID- 10385070 TI - Primary sclerosing cholangitis in children. AB - Primary sclerosing cholangitis (PSC), a chronic inflammatory process affecting the extrahepatic and/or medium to large bile ducts, is not rare in children. It has features suggesting an autoimmune pathogenesis, although the mechanism of tissue damage remains unknown. The clinical presentation of childhood primary sclerosing cholangitis is highly variable and frequently without obvious features of cholestasis. Clinical similarity to autoimmune hepatitis is common. Association with chronic colitis is less common than in adults. Cholangiography is essential for the diagnosis and examination of the medium to large intrahepatic ducts is mandatory, as 40% of children lack extrahepatic duct involvement. Histological findings may help to distinguish childhood PSC from autoimmune hepatitis. In children, sclerosing cholangitis may also develop secondary to other disease processes, notably Langerhans histiocytosis, congenital immunodeficiencies and cystic fibrosis. Neonatal sclerosing cholangitis is chronic inflammatory disease of bile ducts which presents initially with neonatal cholestasis; its pathogenesis remains uncertain and may not be the same as for primary sclerosing cholangitis. Effective treatment modalities for childhood PSC remain undetermined. Liver transplantation is required for children who progress to biliary cirrhosis and hepatic decompensation. PMID- 10385071 TI - Progressive familial intrahepatic cholestasis. AB - Progressive familial intrahepatic cholestasis (PFIC), also known as Byler disease, is an inherited disorder of childhood in which cholestasis of hepatocellular origin often presents in the neonatal period and leads to death from liver failure before adolescence. The pattern of appearance of affected children within families is consistent with autosomal recessive inheritance. Several studies have provided support for the heterogeneity of this clinical entity suggesting the existence of different types due to different disorders affecting the hepatocyte and related to defects of bile acid secretion or bile acid metabolism. Recent molecular and genetic studies have identified genes responsible for three types of PFIC and have shown that PFIC was related to mutations in hepatocellular transport system genes involved in bile formation. These findings now provide specific diagnostic tools for the investigation of children with PFIC and should allow prenatal diagnosis in the future. Genotype phenotype correlations performed in patients treated with ursodeoxycholic acid or biliary diversion should allow those PFIC patients who could benefit from these therapies to be precisely identified. In the future, other therapies, such as cell and gene therapies, might be considered and could also represent an alternative to liver transplantation. PMID- 10385073 TI - Hepatobiliary and pancreatic: cystic liver lesion in a man with abdominal pain. PMID- 10385072 TI - Cardiopulmonary dysfunction in cirrhosis. AB - Cirrhosis is associated with several circulatory abnormalities. These include hyperkinetic systemic and splanchnic circulation, hepatopulmonary syndromes including pulmonary hypertension, and cirrhotic cardiomyopathy. Hepatopulmonary syndrome generally refers to hypoxaemia seen in patients with chronic liver disease and appears to be relatively common, although often subclinical. However, significant pulmonary hypertension occurs in 0.2-0.7% of cirrhotic patients. Nitric oxide and/or other vasodilators appear to be involved in the pathogenesis of hepatopulmonary syndrome through induction of pulmonary capillary dilatation which increases the alveolar-arterial oxygen gradient. Cirrhotic cardiomyopathy refers to abnormal left ventricular function which is manifested under conditions of physiological or pharmacological stress. The emergence of liver transplantation as an effective treatment for end-stage liver disease has led to recognition of previously subclinical cardiomyopathy and congestive heart failure accounts for significant morbidity and mortality after this procedure. Diminished myocardial beta-adrenergic receptor function has been shown to play an important role in the pathogenesis of this condition. The contributions of other factors including nitric oxide, catecholamines and membrane fluidity changes are under investigation. Cirrhotic patients also have an increased incidence of other cardiac abnormalities, such as endocarditis and pericardial effusions. PMID- 10385074 TI - Gastrointestinal: upper oesophageal web. PMID- 10385075 TI - Screening by tRNA primer extension analysis of porcine kidney mRNA libraries defines a novel endogenous porcine retroviral long terminal repeat. AB - BACKGROUND: We have taken advantage of the common requirement of all eukaryotic retroelements for a specific tRNA primer to initiate DNA synthesis and applied a previously described in vitro screening methodology to the analysis of in vivo porcine tissues for transcriptionally active retroviral sequences. METHODS: A series of 18-base pair (bp) 3' tRNA oligomers complementary to established primer binding sites for a variety of vertebrate retroviruses, retrotransposons, and retroposons were applied to primer extension analysis of kidney poly(A) mRNA. Primer extension products are predicted to represent "strong stop" signals characteristic of the initial stages of retroviral transcription. RESULTS: Several extension products were cloned, sequenced, and analyzed as probes for screening the porcine genome for potentially active retroviral sequences. We used this strategy to identify and clone a 655-bp 5' long terminal repeat of a porcine retrovirus with significant homology to the simian sarcoma virus. This transcriptionally active virus has an 82-bp U5 region, a conserved AATAAA polyadenylation sequence, a 39-bp repeat reminiscent of other retroviral enhancers, and a unique glycine primer binding site. CONCLUSION: Our results suggest that tRNA primer cloning can effectively identify novel retroviral elements. PMID- 10385076 TI - Elimination of Kupffer cells and nafamostat mesilate rinse prevent reperfusion injury in liver grafts from agonal non-heart-beating donors. AB - BACKGROUND: We hypothesized that microcirculatory disturbance was an obstacle to liver transplantation (LTx) from non-heart-beating donors (NHBDs) and that it was attributed mainly to a deterioration of sinusoidal endothelial cells (SECs) and sinusoidal narrowing. This study was designed to examine porcine orthotopic LTx using livers obtained from pretreated agonal NHBDs, and to determine whether the maintenance of the liver microcirculation would result in successful LTx from agonal NHBDs. METHODS: Pigs were allocated to five groups: (i) control group; (ii) NM group, in which grafts were rinsed with nafamostat mesilate (NM) rinse; (iii) LD group, in which Kupffer cells in grafts were eliminated by liposome encapsulated dichloromethylene diphosphonate (L-DMDP); (iv) LDNM group, in which grafts pretreated with L-DMDP were rinsed with NM rinse; (v) heart-beating donor (HBD) group. In all groups, but the HBD group, the livers were pretreated with FK506 and prostaglandin I2 analogue, and were preserved in University of Wisconsin solution after cardiac arrest. Thereafter orthotopic LTx was performed. RESULTS: After reperfusion, it was histologically demonstrated that elimination of Kupffer cells prevented SECs deterioration and NM rinse prevented sinusoidal narrowing. The hepatic energy charge recovered in all groups except the control group. In the LDNM group, three of four recipients survived more than 7 days. CONCLUSIONS: For a successful LTx from agonal NHBDs, it is important to prevent microcirculatory disturbance caused by SEC deterioration and sinusoidal narrowing after reperfusion. Combination therapy consisting in the elimination of Kupffer cells and NM rinse prevented primary graft non-function in liver grafts from agonal NHBDs. PMID- 10385077 TI - Evidence for epitope spreading and active suppression in skin graft tolerance after donor-specific transfusion. AB - BACKGROUND: To clarify the controversial results in the literature regarding the role of donor-specific transfusion (DST) on allograft survival, we have examined the influence of the following on DST-induced allograft survival in a 2C transgenic mouse model: varying the time between DST and transplantation; the role of MHC disparities between donor and recipient; whether tolerance induced by DST spreads to skin allografts expressing other alloantigens; and whether cyclosporine (CsA) treatment could further modulate skin allograft tolerance after DST. METHODS AND RESULTS: The studies were performed in both 2C anti-Ld (MHC class I) transgenic and normal (nontransgenic) mice. Our data demonstrate that a single infusion of Ld-mismatched lymphocytes 7 days before transplantation leads to permanent acceptance of donor-specific skin allografts in both transgenic (58/58) and nontransgenic (8/8) mice in the absence of any other nonspecific immunosuppressive treatment. Pretransplantation DST from donors mismatched for more than one MHC antigen (Ag) has no beneficial effect on subsequent donor skin allograft survival. However, Ld plus multiple minor histocompatibility (mH) Ag-mismatched DST induced permanent acceptance of donor specific skin allografts. Tolerance induced by one-locus Ld-mismatched DST spreads to skin allografts expressing either two-locus Ld or one-locus Ld plus multiple mH Ags. Administration of CsA after DST diminished skin allograft survival, rather than enhancing it, suggesting that tolerance in this model system is established by an active immunological process sensitive to CsA. CONCLUSIONS: (1) Pretransplantation infusion of Ld-mismatched lymphocytes in the presence or absence of multiple mH mismatches induces permanent survival of donor specific skin allografts. (2) CsA abrogates DST-induced transplantation tolerance. PMID- 10385078 TI - The evaluation of the safety and tolerability of two formulations of cyclosporine: neoral and sandimmune. A meta-analysis. AB - BACKGROUND: Neoral, a microemulsion formulation of cyclosporine, was approved for use in the United States in 1995. Many studies comparing Neoral and Sandimmune have been conducted, and although most state that Neoral is the superior cyclosporine formulation, results have failed to conclusively demonstrate this claim. The aim of this meta-analysis was to compare the safety and efficacy of Neoral and Sandimmune. METHODS: Publications comparing the use of Neoral and Sandimmune were reviewed for demographic variables, adverse events, rejection incidence, graft losses, and serum creatinine. Neoral and Sandimmune were compared in all patients and in the following subgroups: (1) age (adult or pediatric), (2) transplant type (kidney, liver, or heart), (3) indication (de novo or stable), and (4) study design (randomized prospective trials versus nonrandomized, blinded versus open-labeled studies). RESULTS: The rate of graft loss was similar when comparing Neoral and Sandimmune in all analyses. The incidence of rejection was lower in Neoral-treated de novo renal, liver, and cardiac transplants (P<0.05). There were significantly more adverse events in Sandimmune-treated de novo liver transplants than Neoral-treated de novo liver transplants (P<0.00001). When considering only randomized prospective trials, the incidence of rejection was lower in Neoral-treated de novo and stable patients (P<0.05). However, there were more adverse events in Neoral-treated stable patients (P<0.00001). When considering only blinded studies, there were more adverse events in Neoral-treated patients (P<0.05), whereas in open-labeled studies there was no difference in adverse events comparing Neoral and Sandimmune (P=NS). CONCLUSIONS: Considering all published trials, the data seem to indicate that Neoral therapy is preferred because of a lower rejection incidence, with a trend toward less adverse events. However, when limiting the analysis to only randomized prospective trials, and specifically assessing blinded studies, the data become less clear. Neoral use was associated with more adverse events in blinded studies, and Sandimmune use was associated with more adverse events in open-labeled studies. Careful individual consideration must be given in choosing the best possible cyclosporine formulation. PMID- 10385079 TI - Regulation of the epithelial cell-specific integrin, CD103, by human CD8+ cytolytic T lymphocytes. AB - BACKGROUND: The destruction of the graft epithelium by CD8+ cytolytic T lymphocytes (CTL) is an important aspect of organ allograft rejection. Our recent finding in a mouse model that the epithelial cell-specific integrin, CD103, defines a subset of CD8+ CTL potentially sheds new light onto such interactions. The goal of the present study was to assess the relevance of these data to the human system. METHODS: CD103 expression by human T-cell populations generated in mixed lymphocyte cultures or isolated from transplant nephrectomy specimens was quantitated using multiparameter FACS analyses. RESULTS: CD103 defined a major subset (26-76%) of CD8+ CTL generated in human mixed lymphocyte cultures; cell sorting experiments confirmed that the CD103+ and CD103- subsets both possess allospecific lytic activity. Anti-transforming growth factor (TGF)-beta blocked the appearance of the CD103+ CTL subset, and persistent expression of CD103 by CD8+ CTL was dependent on bioactive TGF-beta. Isolated CD103+ and CD103- CD8 subsets maintained their phenotypic integrity during in vitro expansion, although optimal CD103 expression on the former was TGF-beta dependent. Although CD103+ cells were rare among activated CD8 cells in peripheral lymphoid compartments (< 10%), analyses of transplant nephrectomy specimens revealed that a major subset (21-61%) of CD8 memory/effector cells that infiltrate rejecting renal allografts express high levels of CD103. CONCLUSIONS: We conclude that CD103 defines a discrete and stable subset of human CD8+ CTL and that CD103 expression by such cells is initiated and maintained by bioactive TGF-beta. These data point to the existence of a human effector subset that is uniquely specialized for the destruction of the graft epithelium. PMID- 10385080 TI - Fulminant hepatitis is significantly increased in hepatitis B carriers after allogeneic bone marrow transplantation. AB - BACKGROUND: Bone marrow transplantation (BMT) is effective treatment for many hematologic disease, but performed in a population with a high endemic hepatitis B virus carrier rate, the incidence of liver function impairment and fulminant hepatitis (FH) is expected to be raised. METHODS: Forty-three hepatitis B virus carriers received high-dose chemotherapy and BMT, 32 patients received an allogeneic graft, and 11 patients autologous marrow. Acute graft-versus-host disease prophylaxis consisted of methotrexate on day 1, 3, 6, and 11 and cyclosporine for 6 months. RESULTS: After a median follow-up period of 68 months (range: 1-11.5 years), 26 (81.3%) allogeneic BMT patients developed impaired liver function (LF), 5 progressed to FH on day 93, 169, 170, 180, and 468, respectively, and died after an average of 13.8 days (range: 1-45 days). Whereas only 4 (36.4%) autologous BMT patients developed impaired LF, and none FH. Impaired LF (P=0.026, chi-square), and FH (odds ratio=12.86, P=0.009 for coefficient) were significantly related to an allogeneic marrow graft, and the timing of liver function impairment coincided with cyclosporine withdrawal. Hepatitis B surface antigen (HbsAg) disappeared from the serum in 4/14 (28.6%) patients receiving a marrow graft from an HbsAg+ donor. HbsAg was not detected in the serum after BMT in 2/11 (18.2%) autologous BMT patients. CONCLUSIONS: Hepatitis B virus carriers receiving a marrow graft from an HbsAg+ donor have a significantly increased risk of FH. PMID- 10385082 TI - The incidence and implications of renal cell carcinoma in cadaveric renal transplants at the time of organ recovery. AB - BACKGROUND: With the exception of primary central nervous system tumors, organ recovery is no longer considered from donors with known malignancy. Because intrathoracic and intraabdominal organs are usually recovered before the kidneys, we examined the incidence of renal cell carcinoma in cadaveric donor kidneys at the time of organ recovery. This would establish the theoretical risk of transplanting donor organs from a patient with a known renal malignancy. METHODS: In cooperation with the Louisiana Organ Procurement Agency, we reviewed the records of all patients who were cadaveric kidney donors in the state of Louisiana between September 1991 and October 1997. Information was reviewed and analyzed on donor age, sex, race, past medical/surgical history, cause of death, and the findings at the time of organ recovery. RESULTS: A total of 553 consecutive cadaveric donors were identified, with 1106 kidneys recovered. Of the 553 cadaveric donors, 5 (0.9%) were noted to have an incidental renal cell carcinoma. All tumors were identified in separate donors; that is, none of the tumors were bilateral. None of the five donors had documented symptoms referable to their urinary tract. All tumors were either T1 or T2 by the tumor, node, metastasis classification system, and no evidence of nodal or distant metastatic disease was present. In one case, the contralateral kidney, heart, and liver were transplanted before the tumor was identified. In the remaining four cases, all organs (renal and nonrenal) were discarded. CONCLUSIONS: Renal cell carcinoma is rarely found during renal recovery from a cadaveric donor. However, because the kidneys are usually recovered after the intrathoracic and intraabdominal organs, careful palpation of the kidneys and exploration of any abnormalities is mandated to avoid transplanting any organs from a donor with a known renal malignancy. PMID- 10385081 TI - The results of successful penetrating keratoplasty using donor organ-cultured corneal tissue. AB - BACKGROUND: The aim of this study was identification of predictive factors for postoperative visual acuity in patients with a clear organ-cultured graft and to analyze the change in visual acuity between 12 and 24 months after transplantation. METHODS: The study design was a prospective cohort study. A total of 342 consecutive penetrating keratoplasties using donor organ-cultured grafts, performed in 324 patients, were included. Visual acuity, graft thickness, and graft endothelial cell density were recorded in patients with clear transplants. RESULTS: At 24 months postoperatively, 25 (18.7%) of 134 patients had 20/200 or worse visual acuity and 66 (49.3%) had 20/40 or better visual acuity. Graft thickness took 1 month to decrease to normal values. A temporary graft thinning occurred at 6 months postoperatively, followed by recovery of normal graft thickness by 18 months. The average postoperative endothelial cell density was 1,533+/-598 cells/mm2 during the second year. The 24-month LogMAR (logarithm of minimal angle of resolution) visual acuity correlated with preoperative LogMAR visual acuity (beta=0.26, P=0.005), postoperative lens status (beta=-0.34, P=0.008), preoperative intraocular pressure (beta=0.50, P=0.020), and postoperative astigmatism (beta=0.17, P=0.040). Visual acuity (P=0.022) significantly improved between 12 and 24 months. Preoperative diagnosis (P < 0.0001) and postoperative lens status (P < 0.0001) significantly influenced the change in LogMAR visual acuity between 12 and 24 months. CONCLUSIONS: Donor variables do not influence the visual acuity results of penetrating keratoplasty using organ-cultured donor tissue, whereas they have a strong influence on graft survival and graft endothelial cell density. Visual acuity improves during the first 2 years after transplantation. After keratoplasty, organ-cultured corneal grafts undergo dramatic modifications of their thickness and probably of their transparency. PMID- 10385083 TI - Increased usage of TCR V-beta8 in kidney transplant recipients with aberrant immune reconstitution and clinical complications. AB - BACKGROUND: The efficiency of immunosuppressive drugs prescribed after organ transplantation is mostly monitored through clinical and biological signs of organ rejection or infection. However, it may be expected that some patients develop subtle alterations of their reconstituting immune system, not immediately associated with clinical events. Identification of such anomalies could be useful to alert clinicians for possible future complications. METHODS: A systematic follow-up of peripheral lymphocyte subsets, performed in a cohort of 89 kidney transplant recipients, identified severely skewed CD4/CD8 ratios in 32 patients. For 19 patients, the expression of specific T cell receptor fragments was examined using a panel of 10 monoclonal antibodies. Abnormal control of spontaneously Epstein Barr virus-infected B cells was tested by investigating for the generation of spontaneous lymphoblastoid cell lines in 17 cases. The incidence of rejection and infectious episodes was monitored. RESULTS: A bias in T cell receptor fragments usage was detected in 14/19 cases, involving Vbeta8 in all cases. Spontaneous lymphoblastoid cell lines of Epstein Barr positive B blasts developed in 9 of 17 cases. Eleven patients had early rejection episodes and 16 presented with viral primo-infection or reactivation. The incidence of rejection and infectious episodes was higher in the group of 32 patients who developed such abnormal patterns than in the 57 who did not. CONCLUSION: Transient bias in the T cell receptor repertoire may be observed during immune reconstitution after kidney transplantation, perhaps related to abnormal lymphocyte functions and associated to an impaired control of rejection and/or infectious agents. PMID- 10385084 TI - Modification of glyceraldehyde-3-phosphate dehydrogenase in response to nitric oxide in intestinal preconditioning. AB - BACKGROUND: Previous studies have demonstrated that intestinal preconditioning is triggered by an initial increase in nitric oxide synthesis. This confers resistance to the organ in face of a subsequently sustained period of ischemia reperfusion. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a key enzyme in the glycolytic cascade that could be modulated by nitric oxide. The purpose of the present study is to evaluate a possible inhibitory effect on intestinal GAPDH by the nitric oxide generated during preconditioning. This could lead to a reduction of lactate accumulation during subsequent ischemia. METHODS: GAPDH activity was measured after intestinal preconditioning, and the effect of nitric oxide synthase inhibition was evaluated. RESULTS: Preconditioning induced a significant, but transient, decrease in GAPDH activity. This effect appears to be correlated with a reduced amount of lactate accumulation during ischemia. Inhibition of nitric oxide synthesis reversed these changes. In addition, increased synthesis of nitric oxide was detected after preconditioning. CONCLUSIONS: In summary, this study indicates that nitric oxide generated during ischemic preconditioning could act as a glycolytic modulator during subsequent ischemia, through its effect on GAPDH activity. PMID- 10385085 TI - Interleukin-12 (IL-12)-driven alloimmune responses in vitro and in vivo: requirement for beta1 subunit of the IL-12 receptor. AB - BACKGROUND: Interleukin-12 (IL-12) mediates its biologic activities via binding high-affinity receptors on T and natural killer cells. Although emphasis has been placed on the requirement for IL-12Rbeta2 in IL-12 bioactivity, the role of IL 12Rbeta1 is less well defined. The current study evaluated the effects of exogenous IL-12 on alloantigen-specific immune responses and determined the requirement for IL-12Rbeta1 in IL-12-mediated alloimmunity. METHODS: The mouse heterotopic cardiac transplant model was employed to evaluate the effects of IL 12 on alloantigen-specific immune responses in vivo. In addition, IFN-gamma production in mixed lymphocyte cultures (MLC) supplemented with IL-12 was measured to assess the effects of IL-12 on Th1 function in vitro. Mice deficient in IL-12Rbeta1 (IL-12Rbeta1-/-) were used to determine the requirement for this receptor component in IL-12-driven alloimmune responses. RESULTS: Addition of IL 12 to MLC consisting of wild-type splenocytes enhanced alloantigen-specific proliferative responses and Th1 development. In contrast, IL-12 did not alter these in vitro immune parameters in IL-12Rbeta1-/- MLC. Treatment of wild-type cardiac allograft recipients with IL-12 resulted in high concentrations of serum interferon-gamma (IFN-gamma) and a 10-fold increase in IFN-gamma production by recipient splenocytes after restimulation in vitro. However, this fulminate Th1 response did not accelerate allograft rejection. Importantly, IL-12 had no effect on serum IFN-gamma or in vivo priming of Thl in IL-12Rbeta1-/- recipients. Finally, administration of IL-12 to WT allograft recipients resulted in a bimodal alloantibody response: antibody production was suppressed at high doses of IL-12, and enhanced at lower doses. CONCLUSIONS: IL-12 markedly enhances alloantigen specific immune function; however, these exaggerated Th1-driven responses do not culminate in accelerated allograft rejection. Further, these data indicate that IL-12Rbeta1 is essential for the enhancement of both in vitro and in vivo alloimmune responses by exogenous IL-12. PMID- 10385086 TI - Dominant negative suppression of major histocompatibility complex genes occurs in trophoblasts. AB - BACKGROUND: Polymorphic class I and II major histo-: compatibility complex (MHC) genes are not transcribed in trophoblasts although many immune system cells express these genes constitutively. To study the molecular biology of MHC suppression for the purposes of potential transgenic animal development, we examined the effect on MHC expression in B cells by fusing them with trophoblasts. METHODS: Trophoblasts and B cells with separate selection markers were fused with polyethylene glycol. After growth in double selection media, the hybrids were analyzed for HLA-A, -B, -C, -DR, -DP, and -DQ expression by fluorescence-activated cell scanning and class I and II mRNA by Northern blotting. Class II promoter activity in trophoblasts was then analyzed by transfection of a lethal reporter construct and subsequently, the class II transactivator. RESULTS: Class I and II surface antigens and their corresponding mRNA were completely suppressed in the hybrids. The lethal reporter construct demonstrated that class II suppression resulted from lack of activation of the class II promoter. This in turn was caused by lack of functional class II transactivator. CONCLUSIONS: These data indicate that dominant negative trophoblast factors, either directly or indirectly, suppress expression of the MHC genes. If these factors can be cloned, the potential exists for developing transgenic animals that cannot express MHC or peptide antigen to T cell receptors through the MHC system. PMID- 10385087 TI - Biphasic mechanism for hypothermic induced loss of protein synthesis in hepatocytes. AB - BACKGROUND: A complication in liver transplantation is increased clotting times due to inhibition of protein synthesis resulting from prolonged hypothermic preservation. Protein synthesis is also blocked in cold preserved hepatocytes. In this study, the mechanism of inhibition of protein synthesis in cold preserved hepatocytes was investigated. METHODS: Hepatocytes prepared from rat liver were cold preserved in University of Wisconsin solution for 4, 24, and 48 hr. Protein synthesis was measured as incorporation of radiolabeled leucine into acid precipitable proteins. Hepatocytes were treated with antioxidants (dithiothreitol, trolox or deferoxamine, nitric oxide synthase inhibitor (N(G) monomethyl-L-arginine monoacetate), steroids (dexamethasone or methylprednisolone), methods to keep adenosine triphosphate high (aerobic storage), and cytoskeletal disrupting agents (cytochalasin D or colchicine). RESULTS: There was a 26% decrease in protein synthesis after only 4 hr of cold storage and a further 25% decrease at 24 hr. Antioxidants, elevated adenosine triphosphate, and N(G)-monomethyl-L-arginine monoacetate did not affect the rate of loss of protein synthesis. Protein synthesis was not due to inhibition of amino acid transport or lack of amino acids in the storage medium. Steroid pretreatment of hepatocytes had no effect on the loss of protein synthesis occurring in the first 4 hr of storage but did suppress the loss occurring during the next 44 hr of storage. Cytoskeletal disrupting agents, added to freshly isolated cells, inhibited protein synthesis. CONCLUSION: The mechanism of loss of protein synthesis in cold preserved liver cells is not mediated by: (1) oxygen free radical generation or improved by antioxidant therapy, (2) nitric oxide generation in hepatocytes, (3) an adenosine triphosphate-sensitive destruction of cell viability, and (4) decreased permeability of amino acids or loss of amino acids from the cells. Loss of protein synthesis due to hypothermic storage appears biphasic. The first phase, occurring within 4 hr of storage, may be the result of the effects of hypothermia on the cell cytoskeletal system and may be untreatable. The second phase, which occurs during the next 24 to 48 hr is sensitive to steroid pretreatment. This phase may be amenable to improved preservation methodology. Improved preservation of the liver may require the use of steroids to conserve protein synthetic capabilities. PMID- 10385089 TI - Pretransplant chemotherapy reduces inflammatory cytokine production and acute graft-versus-host disease after allogeneic bone marrow transplantation. AB - BACKGROUND: Tumor necrosis factor-alpha (TNF-alpha) is known to be a critical effector molecule in the pathogenesis of graft-versus-host disease (GVHD), and elevated levels during bone marrow transplantation (BMT) conditioning are associated with more severe GVHD. Many patients receive chemotherapy prior to BMT, but its effect on subsequent toxicities is controversial. METHODS: We studied the effect of prior chemotherapy on GVHD severity and inflammatory cytokine generation in a well-established murine model of allogeneic BMT (B6- >B6D2F1). RESULTS: Three weeks after a single dose of cyclophosphamide, bone marrow and splenic cellularity was reduced by 50% and the production of TNF-alpha to LPS stimulation by macrophages was also markedly impaired (both before and after total body irradiation). Allogeneic BMT recipients previously treated with cyclophosphamide had significantly less GVHD and improved survival relative to recipients previously pretreated with diluent only. This survival advantage was associated with reduced systemic levels of both TNF-alpha and interleukin-1beta 7 days after BMT. This reduction occurred despite equivalent serum levels of lipopolysaccharide, consistent with the reductions in TNF-alpha and interleukin 1beta production by host macrophages after cyclophosphamide pretreatment. CONCLUSIONS: These data support the notion that patients entering BMT conditioning without prior cytotoxic treatment (e.g., patients with chronic myeloid leukemia) may be at increased risk of posttransplant complications associated with excessive inflammatory cytokine production. PMID- 10385088 TI - Pretreatment of crude pancreatic islets with mitomycin C prolongs graft survival time in xenogeneic rat-to-mouse model. AB - BACKGROUND: Rejection of pancreatic islet grafts is still a serious problem. We evaluated the effect of mitomycin C (MMC) on the survival of crude islets grafts after xenogeneic islet transplantation. METHODS: WS (RT1k) rat islets pretreated with various concentrations of MMC (0, 1, 3.2, 10, 32, 50, 100, 320, and 1,000 microg/ml) were transplanted into C57BL/6 mice with streptozotocin-induced diabetes. In vivo graft function was assessed by a daily measurement of nonfasting blood glucose concentration in each animal. We also examined the separate effect of MMC on purified islets and contaminants present in the crude islet preparation. RESULTS: MMC at doses of 10, 32, 50, and 100 microg/ml resulted in a significant prolongation of the mean graft survival time from a control of 12.4+/-2.5 days to 23+/-7.4, 17.5+/-5.4, 25.5+/-14.7, and 26.7+/-8.9 days, respectively. Deterioration of glucose metabolism was noted when the dose exceeded 32 microg/ml, whereas at 320 microg/ ml, MMC failed to restore normoglycemia. Prolongation of survival time of crude islets was the result of its effect on islets and contaminant components of the crude islet preparation. In vitro study showed that MMC treatment at a higher concentration than 10 microg/ml reduces the stimulatory as well as proliferative capacity of lymph node cells. CONCLUSIONS: Pretreatment of pancreatic islets with MMC at 10 microg/ml prolongs xenograft survival without deterioration of in vivo graft function. This novel treatment modality represents a new strategy for the modulation of immunity of islets and contaminants in crude islet preparations. PMID- 10385090 TI - T cells are necessary and critical for xenograft rejection in new concordant cardiac xenotransplant model. AB - BACKGROUND: A new vascularized concordant xenotransplant model using the Chinese hamster as donor and mouse as recipient species is reported. This model takes advantage of the wealth of informative immune reagents and knockout and transgenic backgrounds available for the mouse. METHODS: Heterotopic auxillary cardiac transplantation was performed. The mean survival time was assessed by daily palpation. Xenoreactive antibody production was measured by flow cytometry, and cardiac xenografts were examined by light microscopy. RESULTS: The tempo of xenograft rejection in this model is consistent with concordant species combination. IgM and IgG3 responses were not critical for the concordant xenograft rejection. Long-term survival (>100 days) of the concordant cardiac xenografts was observed without any immunosuppression in nude mice. Reconstitution of nude mice with CD3+ T cells induced the xenograft rejection in 5.7 days (P<0.01). CONCLUSION: This new concordant cardiac xenotransplant model demonstrates that T-dependent xenogeneic immune response is necessary and critical for the xenograft rejection. PMID- 10385091 TI - Magnesium repletion therapy improved lipid metabolism in hypomagnesemic renal transplant recipients: a pilot study. AB - BACKGROUND: Hypomagnesemia has been associated with hypertension, abnormal glucose and lipid metabolism, and accelerated atherosclerosis in nontransplant patients. METHODS: In this prospective short-term pilot study, 14 hypomagnesemic renal transplant recipients with stable renal function were evaluated monthly over a 6-month interval. The first 3 months was the baseline observation period. During the second 3 months, MgO2 was administered to normalize the serum Mg level. Glucose tolerance, lipid levels, blood pressure, weight, and routine chemistries were assessed before and after Mg replacement. All others medications were held constant during the 6-month study. RESULTS: Serum Mg levels increased to normal range after MgO2 therapy, which was well tolerated. There were significant decreases in total cholesterol, low density lipoprotein, and total cholesterol/high density lipoprotein ratio after 3 months of MgO2 therapy. Only three patients had abnormal baseline glucose tolerance tests. All three patients showed improved glucose tolerance after MgO2, but this was not statistically significant. CONCLUSIONS: Mg repletion may be an important ancillary therapy in hypomagnesemic renal transplant patients with hyperlipidemia. PMID- 10385092 TI - Long-term complete remission and immune tolerance after intensive chemotherapy for lymphoproliferative disorders complicating liver transplant. AB - BACKGROUND: B cell lymphoproliferative disorders (LPD) and liver rejection are major lethal complications after hepatic transplantation. Reduction in immunosuppression is the treatment for the former, but is a risk factor for the latter. METHODS: Here, we report three consecutive children with monoclonal LPD complicating orthotopic liver transplantation. All of them were treated with brief (<4 months) but intensive chemotherapy. RESULTS: These three patients have remained in complete remission for LPD for 18 months to more than 3 years. Aggressive antimicrobial prophylaxis was successful in preventing life threatening infections. The patient who received the highest cumulative doses of chemotherapy may have also developed relative immune tolerance to the allograft. CONCLUSIONS: High-dose-intensity chemotherapy may be effective in the treatment of monoclonal LPD, as well as in the induction of immune tolerance for the prevention of allograft rejection and LPD recurrence. PMID- 10385093 TI - Versatility of the inferior epigastric artery as an interpositional vascular graft in living-related liver transplantation. AB - We have used the recipient inferior epigastric artery as an interpositional vascular graft in living-related liver transplantation cases with hepatic artery obstruction, enabling us to restore the arterial inflow sufficiently to the transplanted liver. The inferior epigastric artery is easy to access during abdominal surgery. Easy to harvest, it is anatomically constant and has a caliber equivalent to that of the hepatic artery. Donor site morbidity is negligible. There is no risk of rejection because of the autograft. There has been no report on the availability of the inferior epigastric artery for hepatic artery reconstruction. We consider this vessel as a good option for an arterial conduit in case of the inadequacy or thrombosis of the hepatic artery in living-related liver transplantation. PMID- 10385094 TI - Acute transjugular intrahepatic portosystemic shunt migration into pulmonary artery during liver transplantation. AB - Transjugular intrahepatic portosystemic shunt has become an accepted intervention to treat sequelae of end-stage liver disease such as refractory ascites and esophageal varices for patients awaiting liver transplantation. Technical difficulties in such patients at the time of transplantation are usually limited to malpositioning of the stent requiring modification of the usual vascular anastomoses. Migration of the stent intraoperatively has not been a reported complication in the literature. We report a case in which a patient with a previously placed transjugular intrahepatic portosystemic shunt underwent successful liver transplantation complicated by intraoperative migration of the stent into the left pulmonary artery. The stent was removed from the pulmonary artery postoperatively using interventional radiology techniques. PMID- 10385095 TI - Mycobacterium marinum infection in a renal transplant recipient. AB - BACKGROUND: Infections with atypical mycobacteria occur more frequently in patients with solid organ transplants than in the normal host. METHODS: We report a case of cutaneous Mycobacterium marinum infection in a renal transplant recipient. The patient presented with nodules on the forearm after returning from a fishing trip and was treated for cellulitis without success. RESULTS: Cultures of a biopsy of the lesion grew M. marinum. The patient was treated with ethambutol and ciprofloxacin with a good response; however, 9 months of treatment were required for complete resolution. CONCLUSION: Immunosuppressive therapy for renal transplantation increases susceptibility to a variety of opportunistic infections. A patient who presents with nodules on the extremities should be questioned regarding contact with fish, aquatic environments, or fish tank water, in which case infection with M. marinum should be considered. The diagnosis and treatment of this infection in transplant recipients is discussed. PMID- 10385096 TI - Recurrent cardiocirculatory arrest after kidney transplantation related to intravenous methylprednisolone bolus therapy. AB - BACKGROUND: Intravenous bolus therapy with steroids is often used in standard immunosuppression initially after organ transplantation and to treat acute graft rejection. Although this regimen in generally is safe, severe adverse effects can occur. METHODS: This letter gives a picture of the eventful clinical course of a patient with preexisting heard problems after renal transplantation. RESULTS: This case report proves lethal cardiopulmonary complications closely related to the recurrent intravenous administration of methylprednisolone in a risk patient. CONCLUSIONS: Severe side effects after the application of high-dose steroids are possible. If risk patients are identified, steroid bolus therapy should be avoided or, if not possible, should only be done under close monitoring. PMID- 10385098 TI - Patent rules should include a defence against monopolies. PMID- 10385097 TI - Complement causes allograft injury by cell activation rather than lysis. PMID- 10385099 TI - Boost US infrastructure. PMID- 10385100 TI - Senate seeks $750m for NIH to rebuild ageing biomedical labs. PMID- 10385101 TI - DFG urges Germany to boost its spending on genome research. PMID- 10385102 TI - South African government seeks reassurances on AIDS initiative. PMID- 10385103 TI - Patent on umbilical-cord cells rejected in Europe... PMID- 10385104 TI - As US bid to patent human-animal hybrid fails. PMID- 10385105 TI - Proposed GMO rules lack scientific sense. PMID- 10385106 TI - Others should follow the US line on bioweapons. PMID- 10385107 TI - Cultural primatology comes of age. PMID- 10385108 TI - Enzymes. Picking a winner. PMID- 10385109 TI - DNA repair. Variants on a theme. PMID- 10385110 TI - Comets. Putting the CO in coma. PMID- 10385111 TI - Turning a corner in vision research. PMID- 10385112 TI - Carbohydrate chemistry. Sugars out in the open. PMID- 10385113 TI - Developmental neurobiology. Decoding the Reelin signal. PMID- 10385114 TI - Visual kin recognition in chimpanzees. PMID- 10385115 TI - The oldest fossil ascomycetes. PMID- 10385116 TI - Topography of contextual modulations mediated by short-range interactions in primary visual cortex. AB - Neurons in primary visual cortex (V1) respond differently to a simple visual element presented in isolation from when it is embedded within a complex image. This difference, a specific modulation by surrounding elements in the image, is mediated by short- and long-range connections within V1 and by feedback from other areas. Here we study the role of short-range connections in this process, and relate it to the layout of local inhomogeneities in the cortical maps of orientation and space. By measuring correlation between neuron pairs located in optically imaged maps of V1 orientation columns we show that the strength of local connections between cells is a graded function of lateral separation across cortex, largely radially symmetrical and relatively independent of orientation preferences. We then show the contextual influence of flanking visual elements on neuronal responses varies systematically with a neuron's position within the cortical orientation map. The strength of this contextual influence on a neuron can be predicted from a model of local connections based on simple overlap with particular features of the orientation map. This indicates that local intracortical circuitry could endow neurons with a graded specialization for processing angular visual features such as corners and T junctions, and this specialization could have its own functional cortical map, linked with the orientation map. PMID- 10385117 TI - Identification of two sources of carbon monoxide in comet Hale-Bopp. AB - The composition of ices in comets may reflect that of the molecular cloud in which the Sun formed, or it may show evidence of chemical processing in the pre planetary accretion disk around the proto-Sun. As carbon monoxide (CO) is ubiquitous in molecular clouds, its abundance with respect to water could help to determine the degree to which pre-cometary material was processed, although variations in CO abundance may also be influenced by the distance from the Sun at which comets formed. Observations have not hitherto provided an unambiguous measure of CO in the cometary ice (native CO). Evidence for an extended source of CO associated with comet Halley was provided by the Giotto spacecraft, but alternative interpretations exist. Here we report observations of comet Hale-Bopp which show that about half of the CO in the comet comes directly from ice stored in the nucleus. The abundance of this CO with respect to water (12 per cent) is smaller than in quiescent regions of molecular clouds, but is consistent with that measured in proto-stellar envelopes, suggesting that the ices underwent some processing before their inclusion into Hale-Bopp. The remaining CO arises in the coma, probably through thermal destruction of more complex molecules. PMID- 10385118 TI - Laboratory evolution of peroxide-mediated cytochrome P450 hydroxylation. AB - Enzyme-based chemical transformations typically proceed with high selectivity under mild conditions, and are becoming increasingly important in the pharmaceutical and chemical industries. Cytochrome P450 monooxygenases (P450s) constitute a large family of enzymes of particular interest in this regard. Their biological functions, such as detoxification of xenobiotics and steroidogenesis, are based on the ability to catalyse the insertion of oxygen into a wide variety of compounds. Such a catalytic transformation might find technological applications in areas ranging from gene therapy and environmental remediation to the selective synthesis of pharmaceuticals and chemicals. But relatively low turnover rates (particularly towards non-natural substrates), low stability and the need for electron-donating cofactors prohibit the practical use of P450s as isolated enzymes. Here we report the directed evolution of the P450 from Pseudomonas putida to create mutants that hydroxylate naphthalene in the absence of cofactors through the 'peroxide shunt' pathway with more than 20-fold higher activity than the native enzyme. We are able to screen efficiently for improved mutants by coexpressing them with horseradish peroxidase, which converts the products of the P450 reaction into fluorescent compounds amenable to digital imaging screening. This system should allow us to select and develop mono- and di oxygenases into practically useful biocatalysts for the hydroxylation of a wide range of aromatic compounds. PMID- 10385119 TI - Cultures in chimpanzees. AB - As an increasing number of field studies of chimpanzees (Pan troglodytes) have achieved long-term status across Africa, differences in the behavioural repertoires described have become apparent that suggest there is significant cultural variation. Here we present a systematic synthesis of this information from the seven most long-term studies, which together have accumulated 151 years of chimpanzee observation. This comprehensive analysis reveals patterns of variation that are far more extensive than have previously been documented for any animal species except humans. We find that 39 different behaviour patterns, including tool usage, grooming and courtship behaviours, are customary or habitual in some communities but are absent in others where ecological explanations have been discounted. Among mammalian and avian species, cultural variation has previously been identified only for single behaviour patterns, such as the local dialects of song-birds. The extensive, multiple variations now documented for chimpanzees are thus without parallel. Moreover, the combined repertoire of these behaviour patterns in each chimpanzee community is itself highly distinctive, a phenomenon characteristic of human cultures but previously unrecognised in non-human species. PMID- 10385120 TI - Auditory cortical responses in the cat to sounds that produce spatial illusions. AB - Humans and cats can localize a sound source accurately if its spectrum is fairly broad and flat, as is typical of most natural sounds. However, if sounds are filtered to reduce the width of the spectrum, they result in illusions of sources that are very different from the actual locations, particularly in the up/down and front/back dimensions. Such illusions reveal that the auditory system relies on specific characteristics of sound spectra to obtain cues for localization. In the auditory cortex of cats, temporal firing patterns of neurons can signal the locations of broad-band sounds. Here we show that such spike patterns systematically mislocalize sounds that have been passed through a narrow-band filter. Both correct and incorrect locations signalled by neurons can be predicted quantitatively by a model of spectral processing that also predicts correct and incorrect localization judgements by human listeners. Similar cortical mechanisms, if present in humans, could underlie human auditory spatial perception. PMID- 10385121 TI - The SIL gene is required for mouse embryonic axial development and left-right specification. AB - The establishment of the main body axis and the determination of left-right asymmetry are fundamental aspects of vertebrate embryonic development. A link between these processes has been revealed by the frequent finding of midline defects in humans with left-right anomalies. This association is also seen in a number of mutations in mouse and zebrafish, and in experimentally manipulated Xenopus embryos. However, the severity of laterality defects accompanying abnormal midline development varies, and the molecular basis for this variation is unknown. Here we show that mouse embryos lacking the early-response gene SIL have axial midline defects, a block in midline Sonic hedgehog (Shh) signalling and randomized cardiac looping. Comparison with Shh mutant embryos, which have axial defects but normal cardiac looping, indicates that the consequences of abnormal midline development for left-right patterning depend on the time of onset, duration and severity of disruption of the normal asymmetric patterns of expression of nodal, lefty-2 and Pitx2. PMID- 10385122 TI - Regions of variant histone His2AvD required for Drosophila development. AB - One way in which a distinct chromosomal domain could be established to carry out a specialized function is by the localized incorporation of specific histone variants into nucleosomes. H2AZ, one such variant of the histone protein H2A, is required for the survival of Drosophila melanogaster, Tetrahymena thermophila and mice (R. Faast et al., in preparation). To search for the unique features of Drosophila H2AZ (His2AvD, also referred to as H2AvD) that are required for its essential function, we have performed amino-acid swap experiments in which residues unique to Drosophila His2AvD were replaced with equivalently positioned Drosophila H2A.1 residues. Mutated His2AvD genes encoding modified versions of this histone were transformed into Drosophila and tested for their ability to rescue null-mutant lethality. We show that the unique feature of His2AvD does not reside in its histone fold but in its carboxy-terminal domain. This C-terminal region maps to a short alpha-helix in H2A that is buried deep inside the nucleosome core. PMID- 10385124 TI - The XPV (xeroderma pigmentosum variant) gene encodes human DNA polymerase eta. AB - Xeroderma pigmentosum variant (XP-V) is an inherited disorder which is associated with increased incidence of sunlight-induced skin cancers. Unlike other xeroderma pigmentosum cells (belonging to groups XP-A to XP-G), XP-V cells carry out normal nucleotide-excision repair processes but are defective in their replication of ultraviolet-damaged DNA. It has been suspected for some time that the XPV gene encodes a protein that is involved in trans-lesion DNA synthesis, but the gene product has never been isolated. Using an improved cell-free assay for trans lesion DNA synthesis, we have recently isolated a DNA polymerase from HeLa cells that continues replication on damaged DNA by bypassing ultraviolet-induced thymine dimers in XP-V cell extracts. Here we show that this polymerase is a human homologue of the yeast Rad30 protein, recently identified as DNA polymerase eta. This polymerase and yeast Rad30 are members of a family of damage-bypass replication proteins which comprises the Escherichia coli proteins UmuC and DinB and the yeast Rev1 protein. We found that all XP-V cells examined carry mutations in their DNA polymerase eta gene. Recombinant human DNA polymerase eta corrects the inability of XP-V cell extracts to carry out DNA replication by bypassing thymine dimers on damaged DNA. Together, these results indicate that DNA polymerase eta could be the XPV gene product. PMID- 10385123 TI - G-protein-coupled receptor heterodimerization modulates receptor function. AB - The opioid system modulates several physiological processes, including analgesia, the stress response, the immune response and neuroendocrine function. Pharmacological and molecular cloning studies have identified three opioid receptor types, delta, kappa and mu, that mediate these diverse effects. Little is known about the ability of the receptors to interact to form new functional structures, the simplest of which would be a dimer. Structural and biochemical studies show that other G-protein-coupled receptors (GPCRs) interact to form homodimers. Moreover, two non-functional receptors heterodimerize to form a functional receptor, suggesting that dimerization is crucial for receptor function. However, heterodimerization between two fully functional receptors has not been documented. Here we provide biochemical and pharmacological evidence for the heterodimerization of two fully functional opioid receptors, kappa and delta. This results in a new receptor that exhibits ligand binding and functional properties that are distinct from those of either receptor. Furthermore, the kappa-delta heterodimer synergistically binds highly selective agonists and potentiates signal transduction. Thus, heterodimerization of these GPCRs represents a novel mechanism that modulates their function. PMID- 10385125 TI - A specific partner for abasic damage in DNA. AB - In most models of DNA replication, Watson-Crick hydrogen bonding drives the incorporation of nucleotides into the new strand of DNA and maintains the complementarity of bases with the template strand. Studies with nonpolar analogues of thymine and adenine, however, have shown that replication is still efficient in the absence of hydrogen bonds. The replication of base pairs might also be influenced by steric exclusion, whereby inserted nucleotides need to be the correct size and shape to fit the active site against a template base. A simple steric-exclusion model may not require Watson-Crick hydrogen bonding to explain the fidelity of replication, nor should canonical purine and pyrimidine shapes be necessary for enzymatic synthesis of a base pair if each can fit into the DNA double helix without steric strain. Here we test this idea by using a pyrene nucleoside triphosphate (dPTP) in which the fluorescent 'base' is nearly as large as an entire Watson-Crick base pair. We show that the non-hydrogen bonding dPTP is efficiently and specifically inserted by DNA polymerases opposite sites that lack DNA bases. The efficiency of this process approaches that of a natural base pair and the specificity is 10(2)-10(4)-fold. We use these properties to sequence abasic lesions in DNA, which are a common form of DNA damage in vivo. In addition to their application in identifying such genetic lesions, our results show that neither hydrogen bonds nor purine and pyrimidine structures are required to form a base pair with high efficiency and selectivity. These findings confirm that steric complementarity is an important factor in the fidelity of DNA synthesis. PMID- 10385126 TI - Basis for recognition of cisplatin-modified DNA by high-mobility-group proteins. AB - The anticancer activity of cis-diamminedichloroplatinum(II) (cisplatin) arises from its ability to damage DNA, with the major adducts formed being intrastrand d(GpG) and d(ApG) crosslinks. These crosslinks bend and unwind the duplex, and the altered structure attracts high-mobility-group domain (HMG) and other proteins. This binding of HMG-domain proteins to cisplatin-modified DNA has been postulated to mediate the antitumour properties of the drug. Many HMG-domain proteins recognize altered DNA structures such as four-way junctions and cisplatin-modified DNA, but until now the molecular basis for this recognition was unknown. Here we describe mutagenesis, hydroxyl-radical footprinting and X ray studies that elucidate the structure of a 1:1 cisplatin-modified DNA/HMG domain complex. Domain A of the structure-specific HMG-domain protein HMG1 binds to the widened minor groove of a 16-base-pair DNA duplex containing a site specific cis-[Pt(NH3)2[d(GpG)-N7(1),-N7(2)]] adduct. The DNA is strongly kinked at a hydrophobic notch created at the platinum-DNA crosslink and protein binding extends exclusively to the 3' side of the platinated strand. A phenylalanine residue at position 37 intercalates into a hydrophobic notch created at the platinum crosslinked d(GpG) site and binding of the domain is dramatically reduced in a mutant in which alanine is substituted for phenylalanine at this position. PMID- 10385127 TI - Dorzolamide, visual function and ocular hemodynamics in normal-tension glaucoma. AB - The purpose of this study was to determine how a topical carbonic anhydrase inhibitor, dorzolamide, alters visual function and ocular blood flow in persons with normal-tension glaucoma. Eighteen normal tension glaucoma patients, after washout of other ocular medications, were treated for four weeks with 2% dorzolamide, three times daily. A control group of eleven other normal-tension glaucoma patients received placebo eye drops. Patients were studied before treatment, and after two and four weeks of treatment. Each study included assessment of central visual function (contrast sensitivity), intraocular pressure (IOP), and several aspects of ocular hemodynamics, including measures of retinal arteriovenous passage time, retinal arterial and venous diameters, and flow velocities in the ophthalmic, central retinal, and posterior ciliary arteries. Dorzolamide significantly reduced IOP at two and four weeks (each p<0.01), and at the same time increased contrast sensitivity at both three and six cycles per degree (each p<0.05). Neither of these variables changed significantly in the control group. Dorzolamide also accelerated retinal arteriovenous passage time of fluorescein dye, at constant retinal arterial and venous diameters (p<0.05), but failed to change flow velocities in any retrobulbar vessel. The ability of dorzolamide to improve contrast sensitivity in persons with normal-tension glaucoma may be related to either IOP reduction or altered ocular perfusion. PMID- 10385128 TI - Topical verapamil lowers outflow facility in the rabbit eye. AB - Results of studies examining the mechanism of the ocular hypotensive effect of topical calcium channel blockers are controversial. Whereas evidence obtained in perfused human eyes indicates that these drugs lower intraocular pressure by increasing the aqueous humor outflow, tonographic studies in rabbits have revealed that they reduce both the aqueous humor outflow and inflow. In order to clarify such a discrepancy, the aim of this study was to assess whether the effect of topical verapamil on the facility of aqueous humor outflow in the rabbit eye was dose-related. Total outflow facility was determined by two-level constant pressure perfusion in anesthetized rabbits. The effect of 5 different concentrations on aqueous humor outflow at 60 minutes postdrug was studied in groups of 10 rabbits each. Baseline outflow facility was also determined in a group of 15 rabbits. In order to check the reliability of the method for detecting drug-induced changes in aqueous outflow, the effect of pilocarpine was also tested. Topical verapamil was shown to lower outflow facility in the rabbit eye in a dose-related fashion. On the contrary, topical pilocarpine was found to significantly increase outflow facility. Our data indicate that topical verapamil reduces outflow facility in the rabbit eye. PMID- 10385129 TI - Improvement of ocular blood flow and retinal functions with puerarin analogs. AB - Ischemic retinopathy and, particularly, age-related macular degeneration (AMD) are difficult eye diseases to treat. Since the etiology of these diseases is inadequate blood circulation in the retina and choroid, drugs which can improve blood circulation to these tissues should be beneficial to these diseases. Since fovea is avascular, AMD is closely related to choroidal vascular abnormalities, and drugs which show strong effects to increase choroidal blood flow would be particularly useful. Puerarin and all its derivatives, except ET (puerarin disubstituted with -CH2CH2OH), showed marked increase of choroidal blood flow at various time periods. Even ET showed a tendency to increase choroidal blood flow, though it was not statistically significant. As for b wave recovery, all puerarin analogs showed strong recovery of retinal function after ischemic insult for 30 min. These results indicate that puerarin analogs could be used for the treatment of ischemic retinopathy, and AMD in particular. PMID- 10385130 TI - Effect of topical amosulalol on tissue circulation in the optic nerve head. AB - The effect of topical 0.1% amosulalol on tissue circulation in the albino rabbit optic nerve head (ONH) was investigated using a laser speckle tissue circulation analyzer. Amosulalol was administered into one eye twice daily for 20 days, and vehicle was administered into the other eye in a masked, randomized manner. Intraocular pressure (IOP) was measured every 5 days. The normalized blur value (NB), a quantitative index of tissue blood flow velocity in the ONH, was measured before treatment and 2 hours after the last instillation on day 20. The IOP was also measured at 5-day intervals. Amosulalol decreased IOP by approximately 2 mmHg in the treated eyes (P < 0.01). There was no significant difference in NB between eyes before the first instillation, whereas NB was significantly greater (by approximately 16%) in the amosulalol-treated eye than in the control eye after completion of instillations (P < 0.01). The difference between NB after completion of instillations and that before the first instillation was significantly greater in the ONH of the amosulalol-treated eye than in the contralateral control eye (P < 0.01). Twice-daily instillation of 0.1% amosulalol for 20 days induced a significant increase in tissue blood velocity in the ipsilateral ONH in albino rabbits. PMID- 10385131 TI - Systemic beta-blockade with once daily Betimol or Timoptic-XE. AB - The purpose of this study was to evaluate the effect of timolol hemihydrate 0.5% (Betimol [THH], Ciba Vision Ophthalmics) and timolol maleate gel forming solution 0.5% (Timoptic-XE , [TXE], Merck, Inc.), when both are dosed once daily on the exercise performance. Maximum exercise heart rate reflects systemic beta-blockade activity. Fourteen healthy subjects were randomized to receive either placebo (HypoTears , Ciba Vision Ophthalmics), THH, or TXE by a Latin square technique in a three period crossover design. Subjects were dosed one drop every morning beginning three days before exercise testing. The interval between each test was one week. Exercise testing was performed two hours after dosing. Maximum exercise heart rate showed no statistical difference between TXE and THH (174 +/- 13.1 vs. 172 +/- 14.9 beats/min, respectively, P = 0.72). Both active treatments, however, decreased heart rate compared to placebo (185 +/- 7.3 beats/min, P = 0.017). Time to exhaustion showed no difference between groups (P > 0.10). The double product (product of heart rate and systolic blood pressure) did not show a difference between TXE and THH (P = 0.38) but was reduced compared to placebo (P = 0.0053). One subject on TXE was discontinued from the study after the first exercise test because of secondary heart block during the recovery period. It was concluded that TXE and timolol hemihydrate solution show similar systemic beta-blockade activity when both are dosed once a day. PMID- 10385132 TI - Ginkgo biloba extract increases ocular blood flow velocity. AB - We evaluated a possible therapeutic effect of Ginkgo biloba extract (GBE) on glaucoma patients that may benefit from improvements in ocular blood flow. A Phase I cross-over trial of GBE with placebo control in 11 healthy volunteers (8 women, 3 men: Age; 34 +/- 3 years, mean +/- SE) was performed. Patients were treated with either GBE 40 mg or placebo three times daily orally, for 2 days. Color Doppler imaging (Siemens Quantum 2000) was used to measure ocular blood flow before and after treatment. There was a two week washout period between GBE and placebo treatment. Ginkgo biloba extract significantly increased end diastolic velocity (EDV) in the ophthalmic artery (OA) (baseline vs GBE treatment; 6.5 +/- 0.5 vs 7.7 +/- 0.5 cm/sec, 23% change, p=0.023), with no change seen in placebo (baseline vs GBE-treatment; 7.2 +/- 0.6 vs 7.1 +/- 0.5 cm/sec, 3% change, p=0.892). No side effects related to GBE were found. Ginkgo biloba extract did not alter arterial blood pressure, heart rate, or IOP. Ginkgo biloba extract significantly increased EDV in the OA and deserves further investigation in ocular blood flow and neuroprotection for possible application to the treatment of glaucomatous optic neuropathy as well as other ischemic ocular diseases. PMID- 10385133 TI - Phenylarsine oxide inhibits phosphate uptake in human ciliary non-pigmented epithelial cells. AB - Phenylarsine oxide (PAO), a sulfhydryl modifying reagent and a widely used inhibitor for tyrosine phosphatases and endocytosis, was tested on the level of phosphorylation in human nonpigmented ciliary epithelial ocular (HNPE) cells. Pretreatment with (PAO, 10 microM) for 30 min followed by incubation with 32Pi to stimulate endogenous phosphorylation surprisingly resulted in a total reduction in 32Pi labeled proteins. PAO (10-50 microM) dose-dependently inhibited both sodium-dependent and -independent phosphate uptake in cells. p Hydroxymercuribenzoate (pHMB, 10 microM), another sulfhydryl modifying reagent failed to mimic PAO effects. However, metabolic inhibitors (iodoacetamide (0.1 mM) and 2,4-dinitrophenol (DNP, 0.5 mM) also mimicked PAO effects, suggesting that the inhibition of ATP production may be responsible for attenuation of both phosphate uptake mechanisms. However, sodium-dependent phosphate uptake in isolated plasma membrane vesicles pretreated with PAO was also significantly lower than control vesicles treated with dimethlysulfoxide (DMSO), suggesting that PAO may be directly targeting a component of the sodium-dependent cotransporter. It is suggested that PAO is a novel inhibitor of phosphate uptake in HNPE cells that acts indirectly by inhibiting ATP production and directly by inhibiting the Na-dependent cotransporter. PMID- 10385134 TI - The distribution of gentamicin in the rabbit cornea following iontophoresis to the central cornea. AB - The purpose of this study was to evaluate the penetration of gentamicin into the central, midperipheral and peripheral cornea of rabbits following iontophoresis to the central 3 mm of the cornea. Four groups (groups 1-4) of five rabbits (one eye per rabbit) underwent corneal iontophoresis using gentamicin dissolved in agar. Low (1 mg/ml) and high (10 mg/ml) concentrations of gentamicin in agar were used for one or ten minutes. Two control groups (groups 5 and 6) of five eyes each underwent mock iontophoresis with low and high concentrations of agar gentamicin mixture. Following sacrifice of the rabbits, the central, midperipheral and peripheral parts of each cornea were excised. Gentamicin concentration was determined in each part of every cornea. High concentrations of gentamicin (951.6 +/- 369.4 microg/ml to 26.6 +/- 41.34 microg/ml) were obtained in the central parts of all the iontophoresis-treated corneas. In each group, except group 6, central corneas had higher concentrations of gentamicin compared to midperipheral corneas (p = 0.038 to p = 0.021), and midperipheral corneas had higher levels than peripheral corneas (p = 0.038 to p = 0.021). Following iontophoresis, gentamicin is found in all portions of the corneas; however, the highest concentration of the drug remains in the central cornea. PMID- 10385135 TI - Ocular effects of antimuscarinic compounds: is clinical effect determined by binding affinity for muscarinic receptors or melanin pigment? AB - Although antimuscarinic drugs are being used with increasing frequency in clinical practice for the purposes of mydriasis and cycloplegia, the extent of their actions varies considerably between different compounds. Investigation of the binding characteristics of these agents revealed that as their reported clinical potency increased, so did their specific binding affinity for muscarinic receptors in the iris sphincter and ciliary muscle and their nonspecific binding affinity for melanin pigment. However, the affinity of each drug for melanin pigment was much lower than for the muscarinic receptors. Therefore, although binding to melanin can significantly influence the overall response, differences in the clinical effect of various compounds appear to be primarily due to their differences in specific affinity for muscarinic binding sites. PMID- 10385136 TI - Pharmacological characterization of [3H]-Ifenprodil binding to polyamine binding sites on rabbit and rat retinal homogenates: role in neuroprotection? AB - Polyamine binding sites (PBS) represent one of the modulatory sites on the N methyl-D-aspartate (NMDA) receptor-channel complex. We have characterized [3H] ifenprodil binding to the PBS on washed homogenates of rabbit and rat retinas. Specific binding of [3H]-ifenprodil (2 nM) (in the presence of 3 microM 1,3-Di [2 tolyl] guanidine HCl and 10 microM GBR12909 to block sigma sites) comprised 47 56% of the total binding. Scatchard analyses indicated interaction with apparent high- and low-affinity sites: dissociation constants (K(d)s) = 0.5-0.6 microM and apparent density of sites (Bmax) = 1.5-4.3 pmol/mg protein and K(d)s = 2.0-2.9 microM, and Bmax values = 15.8-17.8 pmol/mg protein (n = 3). Ifenprodil (Ki = 0.4 0.8 microM), eliprodil (Ki = 0.7-0.8 microM), spermine (Ki = 72-79 microM), spermidine (Ki = 283-330 microM), putrescine (Ki > 650 microM) and MK-801 (Ki > 1 mM) (n = 3-5) differentially competed for [3H]-ifenprodil binding. The biphasic competition curves for ifenprodil were resolved into two binding components: rat retinas, IC50high = 0.19 +/- 0.13 microM and IC50low = 8.7 +/- 1.3 microM; rabbit retinas, IC50high = 0.1 +/- 0.01 microM and IC50low = 16.0 +/- 7.8 microM. These studies have shown the presence of specific PBS labeled by [3H]-ifenprodil in the rabbit and rat retinas which may, in part, be responsible for mediating the neuroprotective effects of eliprodil and ifenprodil. PMID- 10385137 TI - The effect of alpha-tocopherol and beta-carotene supplementation on colorectal adenomas in middle-aged male smokers. AB - Epidemiological and experimental studies have indicated that dietary factors such as vitamin C, vitamin E, and beta-carotene are associated with the risk of colorectal cancer. This study was carried out within the Alpha-Tocopherol, Beta Carotene Cancer Prevention Study (ATBC Study), whose participants were randomly assigned to four supplementation groups: (a) alpha-tocopherol (AT), 50 mg/day; (b) beta-carotene (BC), 20 mg/day; (c) both AT and BC; and (d) placebo. We included the 15,538 ATBC Study participants who had been randomized within the areas of three major cities in southern Finland. Cases of colorectal adenoma (n = 146) were identified by the pathology laboratories in the study areas, and these participants' medical records were collected and reviewed. Alpha-tocopherol supplementation increased the risk for adenomas (relative risk, 1.66; 95% confidence interval, 1.19-2.32), whereas beta-carotene supplementation had no effect on the risk (relative risk, 0.98; 95% confidence interval, 0.71-1.35). Slightly more prediagnosis rectal bleeding and intestinal pain occurred in those adenoma cases who received alpha-tocopherol supplements than in those who did not. Thus, some bias may have resulted, with alpha-tocopherol supplementation leading to more colonoscopies and, thus, to an increased detection of incident polyps in this group. This is further supported by the trial finding that alpha tocopherol supplementation did not increase the risk of colorectal cancer. PMID- 10385138 TI - Genetic and dietary predictors of CYP2E1 activity: a phenotyping study in Hawaii Japanese using chlorzoxazone. AB - Cytochrome P4502E1 (CYP2E1) is considered to play an important role in the metabolic activation of procarcinogens such as N-nitrosoamines and low molecular weight organic compounds. An RsaI polymorphism is present in the 5'-flanking region of the CYP2E1 gene, which could possibly affect its transcription. However, the relationship between genotype and the phenotypic catalytic activity of the enzyme has not been defined. Also, the effects in humans of specific dietary factors, other than ethanol, which have been shown in animal and in vitro studies to modulate CYP2E1 activity, are unknown. Accordingly, the CYP2E1 mediated metabolism of chlorzoxazone to its 6-hydroxy metabolite was investigated in 50 healthy Japanese of both sexes in Hawaii. The oral clearance of the in vivo probe, the trait measure of CYP2E1 activity, was smaller than that reported in European-Americans. Significantly, after adjustment for age and sex, the oral clearance of chlorzoxazone decreased with the number of variant c2 alleles, and its mean in the c2/c2 genotype (147 ml/min) was statistically lower (P < or = 0.05) than that for either the homozygous wild-type (238 ml/min) or the heterozygote (201 ml/min) genotypes. Stepwise multiple regression analysis indicated that body weight was a major contributor to the interindividual variability in the oral clearance of chlorzoxazone, accounting for 43% of the variance. Consumption of lettuce, broccoli, and black tea explained additional components of the variability (7, 5, and 6%, respectively), as did medication use (3%), age (4%), and CYP2E1 genotype (5%). Overall, 73% of the variance could be accounted for by these variables. Body weight, lettuce, and use of medications were associated with increased CYP2E1 activity, and the other covariates were associated with reduced enzyme function. Because of the role that CYP2E1 plays in procarcinogen activation, especially of N-nitrosamines involved in lung cancer, the identified factors may account in part for observed differences in individual susceptibility to disease and may also have implications for cancer prevention. PMID- 10385139 TI - Aberrant methylation of p16INK4a in anatomic and gender-specific subtypes of sporadic colorectal cancer. AB - Colorectal cancer (CRC) occurring in the proximal colon and among women may represent a distinct subtype of the disease. In the present study of 120 sporadic CRCs, we used methylation-specific PCR to test whether methylation of the CpG island in the 5' region of the p16INK4a tumor suppressor gene was associated with anatomical location, gender, or other clinicopathological characteristics. Overall, 18.3% of the tumors had detectable p16INK4a methylation. A marked preponderance of methylated tumors occurred within the proximal colon; cancers occurring proximal to the sigmoid colon were 13.1 times more likely to contain methylated p16INK4a compared with distal tumors. In addition, female patients were 8.8 times more likely than males to have methylation-positive cancers, and p16INK4a methylation was also associated with poorly differentiated tumors. The localization of tumors with p16INK4a methylation within the proximal colon and among female patients specifically adds to a growing database of molecular alterations that define important subtypes of sporadic CRC. The potentially reversible nature of CpG methylation may provide novel therapeutic opportunities for this increasing subtype of the disease, which, due to anatomical location, presents a great challenge for early detection. PMID- 10385140 TI - DNA and protein adduct formation in the colon and blood of humans after exposure to a dietary-relevant dose of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine. AB - Epidemiology studies have indicated that certain dietary components, including well-cooked meat, are risk determinants for colon cancer. Cooked meat can contain significant quantities of heterocyclic aromatic amines (HCAs), which have been established as carcinogens in laboratory animals. 2-Amino-1-methyl-6 phenylimidazo[4,5-b]pyridine (PhIP) is usually the most mass-abundant HCA, with concentrations up to 480 ppb. We used accelerator mass spectrometry to establish whether DNA and protein adducts can be detected in humans exposed to a quantity of PhIP comparable with levels of exposure that occur in the diet. Five human volunteers were administered a dietary-relevant dose of [14C]PhIP (70-84 microg) 48-72 h before surgery for removal of colon tumors. Blood samples were collected at various time points, and albumin, hemoglobin, and WBC DNA were extracted for analysis by accelerator mass spectrometry. Tissue samples were collected during surgery and used to assess either tissue available doses of [14C]PhIP or adduct levels. The results of this study show: (a) PhIP is activated to a form that will bind to albumin, hemoglobin, and WBC DNA in peripheral blood. WBC DNA adducts were unstable and declined substantially over 24 h; (b) PhIP is bioavailable to the colon, with levels in normal tissue in the range 42-122 pg PhIP/g tissue; and (c) PhIP binds to both protein and DNA in the colon. DNA adduct levels in the normal tissue were 35-135 adducts/10(12) nucleotides, which was significantly lower than tumor tissue. The results of this study demonstrate that PhIP is bioavailable to the human colon following defined dietary-relevant doses and forms DNA and protein adducts. PMID- 10385141 TI - Methylenetetrahydrofolate reductase, diet, and risk of colon cancer. AB - Individuals with different forms of the 5,10-methylenetetrahydrofolate reductase (MTHFR) gene, carriers of the C677T mutation versus wild type, show differences in enzyme levels; these differences have been hypothesized to be related to DNA methylation and, perhaps, to the nucleotide pool size. Using data from an incident case-control study, we evaluated the combined effect of dietary intake of folate, methionine, vitamin B6, vitamin B12, and alcohol and various forms of the MTHFR gene on risk of colon cancer. Individuals homozygous for the variant form of the MTHFR gene (TT) had a slightly lower risk of colon cancer than did individuals who were wild type [CC, odds ratio (OR) = 0.8, 95% confidence interval (CI) = 0.6-1.1 for men; and OR = 0.9, 95% CI = 0.6-1.2 for women]. High levels of intake of folate, vitamin B6, and vitamin B12 were associated with a 30 40% reduction in risk of colon cancer among those with the TT relative to those with low levels of intake who were CC genotype. Associations were stronger for proximal tumors, in which high levels of intake of these nutrients were associated with a halving of risk among those with the TT genotype. The inverse association with high levels of these nutrients in those with the TT genotype was stronger among those diagnosed at an older age. Although imprecise, the inverse association with the low-risk diet that was high in folate and methionine and without alcohol was observed for both the TT genotype (OR = 0.4 95% CI = 0.1-0.9) and the CC/CT genotype (OR = 0.6, 95% CI = 0.4-1.0), but this association was not seen with the high-risk diet for either the TT or CC/CT genotype. Although associations were generally weak, these findings suggest that those with differing MTHFR genotypes may have different susceptibilities to colon cancer, based on dietary consumption of folate, vitamin B6, and vitamin B12. PMID- 10385142 TI - A case-control study of colorectal adenomatous polyps and consumption of foods containing partially hydrogenated oils. AB - The trans fatty acids produced by partially hydrogenating vegetable oils may cause colorectal neoplasia by interfering with cell membrane function or eicosanoid synthesis. This possibility provides a rationale for looking at the relation between colorectal adenomatous polyps and consumption of foods containing partially hydrogenated vegetable oils (PHVOs). A total of 516 cases and 551 controls who underwent screening sigmoidoscopy from 1991-1993 were recruited from a prepaid Los Angeles health plan. Subjects were interviewed and given a self-administered food frequency questionnaire. Food items containing PHVOs were divided into four groups characterized by principal ingredients and preparation methods: sweetened baked goods, candy bars, oils and condiments, and french fries and chips. After adjusting for age, sex, physical activity, body mass index, smoking, total energy, and red meat and vegetable intake, there was a positive association between polyps and sweetened baked goods [350+ versus <50 kcal/day (odds ratio, 2.1; 95% confidence interval, 1.3-3.5)]. No association was found with the other food groups after adjustment for dietary and nondietary covariates. Neither was total dietary trans fatty acid associated with adenomas after adjustment for sweetened baked goods and other covariates. These results do not support the hypothesis that eating foods containing PHVOs increases the risk of colorectal adenomas, but they are consistent with the hypothesis that foods high in fat and sugar and low in fiber and correlated micronutrients increase the risk of adenomas. PMID- 10385144 TI - Relationships among breast cancer concern, risk perceptions, and interest in genetic testing for breast cancer susceptibility among African-American women with and without a family history of breast cancer. AB - There has been very little research exploring the relationships among perceptions of, and concern about, getting breast cancer and interest in genetic testing for breast cancer among African-American women with and without a family history of breast cancer. This study explored these issues among 130 and 136 African American women with and without a family history of breast cancer, respectively. Women with a family history reported having greater perceived breast cancer risks and concerns than women without a family history of breast cancer. Knowledge of breast cancer risk factors was very poor and correlated weakly with perceptions of risk and concern. In attributional analyses, acknowledging one's family history status was the strongest predictor of perceived risk only among women with a family history. Women with a family history of breast cancer expressed greater interest in genetic testing for breast cancer susceptibility than women without a family history, although interest in testing was high overall. Increasing perceptions of breast cancer risks and concerns were related to a greater interest in genetic testing, and this relationship was not moderated by family history status. Attributions of risk and knowledge of breast cancer risk factors generally were not related to interest in testing. Overall, these results suggest that: (a) African-American women with a family history are more concerned about and do recognize their greater risk of breast cancer; (b) knowledge of risk factors and attributions of risk are not directly related to interest in genetic testing; and (c) concerns, rather than beliefs about one's risk, are more powerfully related to interest in genetic testing, independent of family history status. PMID- 10385143 TI - Serum concentrations of organochlorine compounds and the subsequent development of breast cancer. AB - A nested case-control study was conducted to examine the association between serum concentrations of 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene (DDE), the primary metabolite of 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT), and polychlorinated biphenyls (PCBs) and the development of breast cancer up to 20 years later. Cases (n = 346) and controls (n = 346) were selected from cohorts of women who donated blood in 1974, 1989, or both, and were matched on age, race, menopausal status, and month and year of blood donation. Analyses were stratified by cohort participation because median DDE and PCB concentrations among the controls were 59 and 147% higher in 1974 than 1989, respectively. Median concentrations of DDE were lower among cases than controls in both time periods [11.7% lower in 1974 (P = 0.06) and 8.6% lower in 1989 (P = 0.41)]. Median concentrations of PCBs were similar among cases and controls [P = 0.21 for 1974 and P = 0.37 for 1989 (Wilcoxon signed rank test)]. The risk of developing breast cancer among women with the highest concentrations of DDE was roughly half that among women with the lowest concentrations, whether based on concentrations in 1974 [odds ratio (OR), 0.50; 95% confidence interval (CI), 0.27-0.89; P(trend) = 0.02] or in 1989 (OR, 0.53; 95% CI, 0.24-1.17; P(trend) = 0.08). The associations between circulating concentrations of PCBs and breast cancer were less pronounced but still in the same direction (1974: OR, 0.68; 95% CI, 0.36-12.9; P(trend) = 0.2; and 1989: OR, 0.73; 95% CI, 0.37-1.46; P(trend) = 0.6). Adjustment for family history of breast cancer, body mass index, age at menarche or first birth, and months of lactation did not materially alter these associations. These associations remained consistent regardless of lactation history and length of the follow-up interval, with the strongest inverse association observed among women diagnosed 16-20 years after blood drawing. Results from this prospective, community-based nested case-control study are reassuring. Even after 20 years of follow-up, exposure to relatively high concentrations of DDE or PCBs showed no evidence of contributing to an increased risk of breast cancer. PMID- 10385145 TI - Cervical cancer screening among Cambodian-American women. AB - Southeast Asian women have higher invasive cervical cancer incidence rates and lower Pap testing frequencies than most other racial/ethnic groups in the United States. However, there is little information about the cervical cancer screening behavior of Cambodian-American women. Cambodian residents of Seattle were surveyed in person during late 1997 and early 1998. The PRECEDE model was used to guide the development of items that assessed predisposing, reinforcing, and enabling factors associated with cervical cancer screening participation. The estimated overall survey response was 72%. Four hundred thirteen women completed our questionnaire. Approximately one-quarter (24%) of the respondents had never had a Pap test, and over one-half (53%) had not been screened recently. The following variables were positively associated with a history of at least one Pap smear: younger age, greater number of years since immigration, belief about Pap testing for postmenopausal women, prenatal care in the United States, and physician recommendation. Women who believed in karma were less likely to have ever been screened for cervical cancer than those who did not. Six variables independently predicted recent screening: age; beliefs about regular checkups, cervical cancer screening for sexually inactive women, and the prolongation of life; having a female doctor; and a previous physician recommendation for Pap testing. The study findings indicate that culturally specific approaches might be effective in modifying the cervical cancer screening behavior of immigrant women. Programs targeting Cambodian-Americans are likely to be more effective if they are multifaceted and simultaneously address predisposing, reinforcing, and enabling factors. PMID- 10385146 TI - Glutathione S-transferase M1 and susceptibility to nasopharyngeal carcinoma. AB - Genetic polymorphisms for enzymes that metabolize tobacco smoke have been reported to determine susceptibility to several smoking-related cancers, including cancers of the lung, bladder, and head and neck. Glutathione S transferase M1 (GSTM1) detoxifies benzo(a)pyrene and other carcinogens in tobacco smoke. Approximately 50% of Caucasians lack the GSTM1 gene. Because the most common type of nasopharyngeal cancer (NPC), squamous cell carcinoma, is related to smoking, we sought to determine whether GSTM1 is associated with risk for NPC. Cases (n = 83) were from a population-based study conducted from 1987 to 1993 at five cancer registries in the United States. Random-digit dialing controls (n = 114) were matched to the cases for age, sex, and registry. Subjects participated in a phone interview and blood draw. Absence of GSTM1 was associated with increased risk for NPC (odds ratio = 1.9, 95% confidence interval = 1.0-3.3 for all cases; and odds ratio = 1.7, 95% confidence interval = 0.8-3.5 for squamous cell cases). This relationship was not modified by smoking history, but stronger relationships between glutathione S-transferase and NPC were suggested among subjects who used alcohol more frequently than others, older subjects (50 or more years of age), and women relative to men. These data indicate that absence of GSTM1 moderately increases risk for NPC and add to growing evidence that GSTM1 is a determinant of risk for several smoking-related cancers. PMID- 10385147 TI - Factors that influence the DNA repair capacity of normal and skin cancer-affected individuals. AB - DNA repair capacity (DRC) was studied in 49 patients affected by basal cell carcinoma (BCC) and 68 cancer-free controls belonging to a larger case-control population enrolled for studying BCC risk factors. DRC was measured in the subjects' peripheral blood lymphocytes by using a host-cell reactivation assay that measures cellular activation of a reporter gene irradiated with UV light. A statistically significant age-related decline in DRC was observed in the controls from 20 to 70 years of age but not in the BCC cases. When the DRC values of the BCC patients and controls were compared by age, young BCC cases (age, < or =40 year) repaired less than the controls, although the difference was not statistically significant. Conversely, older BCC patients (age, >40 years) presented an enhanced repair capacity (P < 0.001) as compared with their controls. The search for possible factors associated with the high repair rate of elderly BCC cases revealed that both target cell physiology and life-style habits may affect host DNA repair. Smoking was the variable that explained most of the increase in DRC among older patients. The understanding of how these factors affect host DRC will be relevant for a correct use of this biomarker. PMID- 10385149 TI - Reproducibility and validity of radioimmunoassays for urinary hormones and metabolites in pre- and postmenopausal women. AB - The reproducibility of RIAs of circulating sex hormones has been evaluated as part of recent epidemiological investigations, but none seem to have addressed the reproducibility or validity of RIAs for urinary hormones or their metabolites. As part of a case-control study of breast cancer in Asian-American women, 12-h overnight urine samples were obtained, and a methodological study was conducted to identify laboratories capable of assaying urinary hormones. For the reproducibility component of this study, two laboratories with extensive experience in hormone assays measured urinary estrone, estradiol, estriol, pregnanediol glucuronide, and estrone glucuronide using samples from 15 women (5 midfollicular, 5 midluteal, and 5 postmenopausal). Variance estimates from these measurements were used to calculate the laboratory variability (coefficient of variation) and to assess the magnitude of the biological variability among the women in relation to the total variability (intraclass correlation coefficient). For the validity component, urinary estrone, estradiol, and estriol levels were measured in the same samples by gas chromatography-mass spectroscopy in the laboratory of Dr. Herman Adlercreutz (University of Helsinki, Helsinki, Finland). We found that the degree of assay reproducibility differed between the laboratories, but that laboratory variability was usually low compared with the range of hormone values among women, particularly for the estrogens. Values for estrone and estradiol were well correlated among all of the laboratories. For estriol, the RIAs tended to overestimate levels compared with gas chromatography mass spectroscopy. In one laboratory, assays for pregnanediol glucuronide and estrone glucuronide were consistently reproduced; in the other, the reproducibility of the RIA for pregnanediol glucuronide was problematic, and estrone glucuronide was not measured. Despite some limitations, urinary hormones and their metabolites can be reliably measured by current RIAs in large investigations attempting to link hormone level to disease risk and may be particularly advantageous for studies of postmenopausal women, where serum concentrations of estrone and estradiol are low and assay measurements are not as dependable. PMID- 10385148 TI - Seasonal effect on airborne pyrene, urinary 1-hydroxypyrene, and benzo(a)pyrene diol epoxide-hemoglobin adducts in the general population. AB - Exposure to airborne polycyclic aromatic hydrocarbons (PAHs) in 65 employees (40 sampled both in summer and winter, 15 sampled in summer only, and 10 sampled in winter only) with no occupational exposure to PAHs was assessed by measuring: personal exposure to pyrene, urinary excretion of 1-hydroxypyrene (1-OHP), and benzo(a)pyrene diol epoxide adducts to hemoglobin (BPDE-Hb). Overall, office employees were exposed to significantly higher levels of pyrene in winter (4.54 +/- 2.35 ng/m3, mean +/- SD) than in summer (1.67 +/- 1.92 ng/m3, mean +/- SD; P < 0.001), but no such seasonal variability was observed in 1-OHP excretion. Tobacco smoking was the major determinant of 1-OHP excretion. BPDE-Hb adducts were measured by gas chromatography-mass spectrometry as benzo(a)pyrene tetrols (BPT) released from adducted hemoglobin. In the 65 employees analyzed, mean BPT levels +/- SD were higher in winter (0.14 +/- 0.38 fmol/mg Hb) than summer (0.031 +/- 0.022 fmol/mg Hb). This difference was not statistically significant, probably because of the small proportion of subjects with detectable adducts (11% in summer and 16% in winter). BPDE-Hb adducts were not significantly associated with sex, age, diet, smoking habits, or with pyrene levels and 1-OHP excretion. This is the first report providing reference BPDE-Hb adduct values for the general population not occupationally exposed to environmental PAHs and shows a tendency to seasonal variability, with higher BPT levels in winter when environmental PAHs are also high. PMID- 10385150 TI - Coronary stenting during rescue angioplasty after failed thrombolysis. AB - Compared with primary angioplasty [percutaneous transluminal coronary angioplasty (PTCA)], rescue PTCA is associated with lower angiographic success and higher reocclusion rates, especially after thrombolysis with tissue-type plasminogen activator (tPA). Although stent placement during primary PTCA has been demonstrated to be safe and even to improve the angiographic results achieved by balloon-alone PTCA, there are few data on stent placement during rescue PTCA after failed thrombolysis. This study sought to assess the feasibility and safety of stent implantation during rescue angioplasty in myocardial infarction after failed thrombolysis. The study population consisted of 20 patients with acute myocardial infarction referred for rescue PTCA after failed thrombolysis consecutively treated with coronary stenting. The thrombolytic agent was tPA in 15 patients (75%), streptokinase in 1 (5%), and anisoylated streptokinase plasminogen activator complex (APSAC) in 1 (5%); 3 patients (15%) were included in the INTIME II study (tPA vs. lanoteplase). After stenting, aspirin 200 mg daily plus ticlopidine 250 mg b.i.d. were administered. Thirty stents (1.5+/-1.0 per patient) were implanted. Angiographic success was achieved in 19 patients (95%). Two patients (10%) died, both because of severe bleeding complications. One patient (5%) suffered a reinfarction, but no patients suffered postinfarction angina or needed new target vessel revascularization. Eighteen patients (90%) were discharged alive and free of events. All these patients remained asymptomatic and free of target vessel revascularization at 6-month follow-up. Stent placement during rescue PTCA after failed thrombolysis is feasible and safe and is associated with a good angiographic result and clinical outcome. Bleeding complications seem to be, however, the main limitation of this reperfusion strategy. PMID- 10385151 TI - Evaluation of contrast agents for delineation of vessel wall boundary by intracoronary ultrasound after coronary angioplasty in human. AB - We evaluated the potential for improving visualization at intervention sites using contrast-enhanced intracoronary ultrasound (ICUS) and the suitable contrast agents for this procedure in humans. In 37 patients, ICUS (30 MHz) was performed with intracoronary bolus injection (3 mL) of seven different contrast preparations and without the contrast agents (control) after coronary intervention. The contrast agents used were as follows: saline solution, standard iomeprol, standard ioxaglate, sonicated iomeprol, sonicated ioxaglate, 50% Albunex, and 100% Albunex. Homogeneous and complete opacification of the vessel lumen and false lumen was observed with sonicated ioxaglate, 50% and 100% Albunex. Shadowing was not observed at all with sonicated ioxaglate and was uncommon with 50% Albunex, whereas 100% Albunex caused shadowing in all cases. The coronary delineation rate with the other contrast agents was only 60%-70%, and the homogeneity and peak intensity were relatively low. Thus, sonicated ioxaglate and 50% Albunex both achieved good visualization, but the latter is more expensive, more difficult to handle, and takes longer to prepare. Of the agents we studied, sonicated ioxaglate appears to be best suited for contrast enhanced ICUS. ICUS using suitable contrast agents could only visualize the large dissections and the strategy was changed according to the contrast-enhanced ICUS results in five cases. Thus, suitable contrast agents, e.g., sonicated ioxaglate, should be used during ICUS after intracoronary intervention. PMID- 10385152 TI - Intravascular ultrasound for evaluation of initial vessel patency and early outcome following directional coronary atherectomy. AB - Elastic recoil and thrombus formation may potentially occur following directional coronary atherectomy (DCA) confounding the assessment of late vascular remodeling. Since intravascular ultrasound (IVUS) data on early outcome of DCA is not available, we used IVUS to investigate whether elastic recoil or thrombus formation can affect early (4 hr) outcome. Quantitative coronary angiography (QCA) and IVUS were performed in high-grade coronary lesions in 32 consecutive patients before, immediately after, and 4 hr after DCA. Late clinical follow-up was obtained after a maximum interval of 2 years. Significant acute elastic recoil was observed by both IVUS (19%+/-14%) and QCA (19%+/-12%), but there was no further recoil after 4 hr. DCA reduced plaque area by 51%+/-13%, an effect that was stable after 4 hr, indicating the absence of relevant thrombus formation. Residual area stenosis by IVUS was not related to the occurrence of late clinical events (n = 8). Mechanical recoil or thrombus formation do not hamper initial lumen gain achieved by DCA. Although QCA significantly underestimated residual plaque burden after DCA when compared to IVUS, the degree of residual area stenosis did not identify patients suffering from cardiac events on follow-up. PMID- 10385153 TI - Immediate and mid-term results after MultiLink stent implantation in native coronary arteries. AB - Different stent designs have widely disparate characteristics that may exert a positive or negative impact on their early and mid-term outcomes. The MultiLink stent (Guidant/Advanced Cardiovascular Systems, Santa Clara, CA) is a new coronary stent with only very limited data. In this report, we examined the results of 50 consecutive patients treated with 57 premounted sheathless MultiLink stents in 53 native coronary arteries with reference diameter > or =2.7 mm. Successful stenting was achieved in 98% of patients, resulting in an improvement in diameter stenosis from 91%+/-11% to 1%+/-3% (P = 0.0001). At 1 month, there was no death, myocardial infarction, or stent thrombosis. Angiographic restudy at a mean of 5.0+/-1.8 months in 94% of patients revealed an in-stent restenosis rate of 20.7%. The restenosis rates for diabetic patients (vs. nondiabetic patients), type C lesions (vs. type A/B1 lesions), and the use of 35-mm-long stents (vs. 15-mm-long stents) were 45.4% (14.3%), 56% (< or =11%), and 80% (8.8%), respectively (P < 0.05). In conclusion, the present study demonstrates that the MultiLink stent has an excellent performance profile, is associated with a low risk of stent thrombosis in native coronary vessels, and yields a favorable restenosis rate, particularly after the use of short (15 mm) stents to treat simple lesions. PMID- 10385154 TI - Benefit of intravascular ultrasound in Wiktor stent implantation. AB - The goal of this study was to evaluate retrospectively the efficacy and safety of intravascular ultrasound (IVUS) during Wiktor stent implantation. Until 1996, the success of stent implantation was assessed by angiographic criteria only, but in 1997, the procedure was expanded to include pre- and postprocedural IVUS imaging. Sixty-six patients were included, 28 treated in 1996 (group A) and 38 treated in 1997 (group B). Stent size was larger in group B than in group A (3.6+/-0.4 vs. 3.1+/-0.2, P < 0.001). Acute gain was greater in group B than in group A (2.58+/ 0.61 vs. 2.11+/-0.56, P < 0.001). Restenosis rate was 31% in group A and 14% in group B. No major acute complications due to IVUS examination occurred. IVUS is helpful in choosing optimal stent size. PMID- 10385155 TI - Simultaneous stent implantation for coarctation of the aorta and closure of patent ductus arteriosus using the Amplatzer duct occluder. AB - We report on a 13-year-old girl with coarctation of the aorta and patent ductus arteriosus who underwent successful simultaneous stent implantation for the coarctation and catheter closure of the ductus using an Amplatzer duct occluder. PMID- 10385156 TI - Stents and Amplatzers: what's an interventionalist to do? PMID- 10385157 TI - Transhepatic therapeutic cardiac catheterization: a new option for the pediatric interventionalist. AB - We evaluate the efficacy and safety of percutaneous transhepatic (TH) venous access for interventional cardiac catheterization. A retrospective review of all TH therapeutic catheterizations between January 1994 and September 1998 was performed. Patient demographics, pre- and postcatheterization hemoglobin and liver function studies, and complications were evaluated. TH access was performed for 30 interventional catheterizations in 25 patients with a median age of 39 months (range, 1 day to 41 years) and weight of 13.2 kg (3.1-87.0 kg). Indications for TH access were bilateral obstructed femoral veins (n = 15), obstructed femoral veins and superior vena cava (n = 3), Greenfield filter (n = 2), and presumptive improved route for intervention via TH access (n = 5). TH interventions were successful in 29/30 procedures (97%). Interventions via TH sheath sizes of 4-14 Fr included pulmonary angioplasty +/- stent (n = 11), radiofrequency ablation (n = 4), atrial septal defect device occlusion (n = 2), coil occlusion of pulmonary artery pseduoaneurysm (n = 2), Fontan fenestration device occlusion (n = 2), pulmonary valvuloplasty (n = 2), stent dilation of the superior vena cava (n = 2), and one each of device retrieval, Fontan baffle stent placement and subsequent redilation, Fontan fenestration dilation, transseptal mitral valvuloplasty, and cardiac biopsy. There were no changes in pre- and post TH hemoglobin levels (mean +/- SD, 12.9+/-2.2 vs. 11.9+/-1.9 gm/dL; P = NS) or alanine transferase (34.0+/-27.5 vs. 43.4+/-18.2 IU/L; P = NS). One patient developed important intraperitoneal bleeding and required exploratory laporatomy. Percutaneous TH access is safe and effective as a route for interventional catheter procedures for patients with limited venous access. PMID- 10385158 TI - Transhepatic access: coming of age. PMID- 10385159 TI - Repeat balloon dilation of congenital valvar aortic stenosis: immediate results and midterm outcome. AB - While balloon dilation (BD) has become the initial treatment for congenital valvar aortic stenosis (CVAS) at many institutions, repeat BD for recurrent obstruction has been reported only in a few. Between January 1985 and December 1996, 298 patients (70 neonates) underwent BD, 34 of whom underwent a repeat BD without mortality. A greater proportion of neonates had a repeat BD (26% vs. 8%, P < 0.001). At repeat BD (1 day-7.5 years post initial BD), the mean peak-to-peak gradient was reduced from 67+/-24 to 36+/-16 mm Hg (P < 0.0001). Aortic regurgitation (AR) increased immediately in 26%, being moderate or more in 24%. During a mean follow-up of 5.2 years, there was one surgically related death. Of the 33 survivors, 6 had surgery for residual stenosis and/or AR. Among the remaining 27 patients, 96% were asymptomatic, the peak instantaneous aortic valve Doppler gradient was 50+/-15 mm Hg with AR absent in 8%, mild in 62%, and moderate or more in 31%. In conclusion, repeat BD is effective and without mortality. AR was at least moderate in 24% of patients immediately after a second BD. Repeat BD was more common in patients who underwent the initial BD as neonates. PMID- 10385160 TI - Relief of right ventricular to pulmonary artery conduit stenosis using a self expanding stent. AB - Intravascular stents have recently been used to treat vascular stenoses in congenital heart disease. Size limitations, however, may preclude their use in certain situations. We describe the successful relief of right ventricular to pulmonary artery conduit stenosis in an adult patient late after repair of truncus arteriosus using a larger, self-expanding wall stent. PMID- 10385161 TI - Non-surgical retrieval of a bullet embolus from the right heart. AB - We report on a case of a 0.38 caliber bullet embolizing from the left common iliac vein to the right atrium. The bullet was successfully retrieved with a percutaneous transvenous catheter technique. Prerequisites for missile embolization and principals of management are discussed. PMID- 10385162 TI - An unusual complication during deployment of a Multi-Link stent. AB - The use of intracoronary stents has greatly impacted on the practice of interventional cardiology. Complications due to equipment failure during deployment of stents are rare but potentially serious. We report a case of a malfunctioning Multi-Link delivery system and the successful treatment of the resulting complications. PMID- 10385163 TI - Acute alveolar hemorrhage and orthodeoxia induced by intravenous amiodarone. PMID- 10385164 TI - Platypnea-orthodeoxia syndrome: etiology, differential diagnosis, and management. PMID- 10385165 TI - Left ventricular free wall rupture during coronary intervention after acute myocardial infarction: report of two cases exhibiting fatal pseudocomplications. AB - Two cases of left ventricular free wall rupture occurring in temporal relation to interventional coronary procedures are presented as autopsy-verified pseudocomplications. The possible impact of pseudocomplications on operator specific registry data and credentialing is briefly discussed. PMID- 10385167 TI - Hemodynamic rounds series II: evaluating new hemodynamic criteria of constrictive physiology: respiratory dynamics. PMID- 10385166 TI - Pericardium-covered stent for septal myocardial ablation in hypertrophic obstructive cardiomyopathy. AB - This report describes a patient with severe hypertrophic obstructive cardiomyopathy in New York Heart Association functional class III. Complete reduction of left ventricular outflow tract gradient was achieved by the selective occlusion of three target septal arteries with a pericardium-covered stent. The patient's in-hospital course was uneventful and has improved to functional class I. PMID- 10385168 TI - Cine PACS: preprint. AB - The technology of cardiac image management is increasingly digital. This tutorial introduces some of the basic concepts relevant to the application of this technology in the cardiac catheterization laboratory. PMID- 10385169 TI - Clinical application of a new rheolytic thrombectomy catheter system for massive pulmonary embolism. PMID- 10385170 TI - Rheolytic thrombectomy: a new treatment for stent thrombosis. AB - Coronary stent thrombosis, a rare complication after stent deployment, carries major morbidity and mortality. Traditional treatments for stent thrombosis include local or systemic delivery of thrombolytic agents and balloon angioplasty, both with far from optimum results. We report on two cases of coronary stent thrombosis successfully treated with rheolytic thrombectomy as an adjunct to balloon angioplasty. PMID- 10385171 TI - Local drug delivery: impact of pressure, substance characteristics, and stenting on drug transfer into the arterial wall. AB - Injection parameters for local drug delivery are frequently determined by studies with marker substances. However, the pharmacologic properties of the actual drug may influence delivery efficiency and lead to different results. Aim of this study was to assess the delivery capacities of two device-drug combinations in order to verify this approach for further in vivo studies. Tritiated (3H) preparations (5 ml) of the hydrophylic low-molecular-weight heparin reviparin and the lipophilic taxane paclitaxel were injected into the left anterior descending artery of freshly explanted porcine hearts with the Infusasleeve II catheter system. A balloon support pressure of 6 atm and infusion pressures of 40, 60, 80, or 100 psi were used. In three additional groups, reviparin was injected following stent implantation and paclitaxel was injected prior to or following stent implantation. Arteries along with surrounding myocardium were harvested. The artery was carefully dissected, and artery and myocardium were separately homogenized, and activity was measured. Of the totally delivered activity, 0.09%+/-0.03% (40 psi) to 0.17%+/-0.13% (100 psi) of reviparin and 2.03%+/-0.67% (60 psi) to 2.68%+/-1.57% (100 psi) of paclitaxel were found in the vessel wall. The results for different injection pressures were not significantly different for either drug. The percentage activity delivered to the vessel wall was substantially larger in the paclitaxel group as compared to reviparin delivery (P < 0.01 at 60, 80, and 100 psi). The mean concentration of reviparin in the artery was 20 to 33 times higher than in the myocardium. For paclitaxel the factors were 110 to 243. Stent implantation prior to or following local delivery did not result in a different delivery efficiency. The results demonstrate that the characteristics of the delivered drug contribute largely to the delivery efficiency. Using identical injection parameters, drug concentrations in the arterial wall were significantly higher for the lipophilic paclitaxel as compared to the hydrophilic reviparin. Stenting of the artery did not influence delivery efficiency. PMID- 10385172 TI - The right stuff (to the right place, at the right dose...) PMID- 10385173 TI - Subxyphoid access of the normal pericardium: a novel drug delivery technique. AB - The pericardial space may potentially serve as a drug delivery reservoir that might be used to deliver therapeutic substances to the heart. This study describes a novel delivery technique that enables safe and rapid percutaneous subxyphoid access of the normal pericardium in a large animal model (49 Yorkshire pigs). An epidural introducer needle (Tuohy-17) is advanced gently under fluoroscopic guidance with a continuous positive pressure of 20-30 mm Hg (achieved by saline infusion using an intraflow system). The positive pressure is intended to push the right ventricle (with a lower pressure) away from the needle's path. Entry of the pericardial space is suspected after an increase in the saline flow through the intraflow system. Access to the pericardial space is confirmed by the injection of 1 ml of diluted contrast under fluoroscopy. A soft floppy-tip 0.025" guidewire is then advanced to the pericardial space and the needle is exchanged for an infusion catheter. Access of the pericardial space was achieved in all animals without any adverse events and without any hemodynamic compromise even with the delivery of fluid volumes as large as 50 ml. Histologic examination in 15 animals 4 weeks after pericardial access did not reveal any delivery-related myocardial damage. The safety, ease, and absence of hemodynamic compromise make this technique a potentially useful method for intrapericardial drug delivery and a good alternative to standard pericardiocentesis in patients with small pericardial effusions at higher risk for complications. PMID- 10385174 TI - Is transseptal left heart catheterization for bioprosthetic mitral valve evaluation really necessary? PMID- 10385175 TI - Intravascular ultrasound and electron beam tomography of late developing coronary artery aneurysms after Kawasaki disease. PMID- 10385176 TI - Extreme left atrial enlargement causing upward displacement and compression of the right pulmonary artery. PMID- 10385177 TI - Crosswire for recanalization of total occlusive coronary arteries. PMID- 10385178 TI - Single vs double transseptal puncture for balloon mitral valvuloplasty...there is always a better way! PMID- 10385179 TI - House staff teaching and nuclear cardiology. PMID- 10385180 TI - The prognostic value of a normal Tc-99m sestamibi SPECT study in suspected coronary artery disease. AB - BACKGROUND: Myocardial perfusion is widely used for risk stratification of patients with suspected or known coronary artery disease (CAD). Recent years have seen an increasing demand for screening of such patients. The value of a normal stress thallium-201 scanning is well established. The advent of technetium 99m sestamibi single photon emission computed tomography (SPECT) has enhanced the profile of nuclear cardiology even further as a reliable test for screening. However, in spite of previous reports, there is paucity of large-scale data regarding the prognostic value of a normal Tc 99m-sestamibi scanning result. METHODS: The aim of our study was to assess the incidence of cardiac death and non-fatal myocardial infarction in patients with an intermediate probability of coronary artery disease (CAD). A total of 473 patients with normal stress Tc-99m sestamibi SPECT were monitored for 30+/-16 (6 to 56) months to assess serious cardiac events. There were 272 men and 201 women, with a mean age of 56+/-2 years, of whom 89% had symptoms suggestive of CAD, 65% had an abnormal exercise electrocardiography, 6% had known CAD, and 5% had a high risk of CAD. The average workload was 9.14 metabolic equivalents, peak exercise heart rate was 93%+/-13% of the age predicted target. RESULTS: The annualized mortality rate was 0.2% (95%CI 0.02% to 0.7%) and no infarctions occurred in this group. CONCLUSIONS: A normal stress Tc-99m-sestamibi is highly predictive of a benign outcome, even in patients with intermediate probability of CAD. PMID- 10385181 TI - Interpretive reproducibility of stress Tc-99m sestamibi tomographic myocardial perfusion imaging. AB - BACKGROUND: Observer variability has been shown with interpretation of planar thallium-201 images. The interpretive reproducibility of technetium-99m sestamibi tomographic imaging is unknown. This study evaluated the interpretive reproducibility of interpretable Tc-99m sestamibi tomographic images among nuclear cardiologists with a wide range of training and experience. METHODS: Three experienced readers (EX) and 3 less-experienced readers (LEX) interpreted 138 exercise and rest Tc-99m sestamibi tomographic images (101 were abnormal in patients with coronary artery disease [CAD], 37 were normal in patients with <5% likelihood of CAD) twice in random sequence without clinical data. Images of good to excellent quality were randomly selected from a database at 2 nuclear cardiology laboratories. Intraobserver and interobserver agreement for global, left anterior descending (LAD) territory, non-LAD first (normal/abnormal) and second (normal/fixed/reversible) order, and defect extent (normal/single-vessel CAD/multi-vessel CAD) were assessed with percent agreement and Cohen's kappa (kappa) statistic. RESULTS: With regard to intraobserver agreement, first and second order ranged from 87% to 94% and 80% to 90% for global, 82% to 96% and 78% to 95% for LAD, and 88% to 91% and 80% to 90% for non-LAD, respectively. Defect extent ranged from 75% to 90%. There were no differences between EX and LEX for global and non-LAD first and second order, LAD first order, and defect extent. LAD second order was 93% for EX compared with 88% (P = .015) for LEX. With regard to interobserver agreement, first and second order ranged from 73% to 89% and 64% to 85% for global, 73% to 93% and 69% to 91% for LAD, and 76% to 88% and 68% to 84% for non-LAD, respectively. Defect extent ranged from 61% to 82%. Global first and second order ranged from 85% to 87% and 78% to 82% for EX compared with 73% to 84% and 64% to 79% for LEX. LAD first and second order ranged from 89% to 91% and 88% to 89% for EX compared with 73% to 91% and 69% to 70% for LEX. Non-LAD first and second order ranged from 82% to 86% and 76% to 77% for EX compared with 76% to 86% and 68% to 81% for LEX. Defect extent ranged from 69% to 75% for EX compared with 59% to 77% for LEX. CONCLUSIONS: There is moderate to excellent interpretive reproducibility with stress Tc-99m sestamibi SPECT imaging among nuclear cardiologists with a wide range of training and experience. PMID- 10385182 TI - Exercise-test Tc-99m tetrofosmin SPECT in patients with chronic ischemic left ventricular dysfunction: direct comparison with Ti-201 reinjection. AB - BACKGROUND: This study was designed to compare the results of exercise-rest technetium-99m tetrofosmin single photon emission computed tomography (SPECT) with those of thallium-201 reinjection at rest after exercise-redistribution imaging in the same patients with chronic ischemic left ventricular (LV) dysfunction. METHODS: Within 1 week, 33 patients with chronic myocardial infarction and LV dysfunction underwent exercise-rest tetrofosmin SPECT and Tl 201 reinjection at rest after exercise-redistribution imaging. In each patient, regional tetrofosmin and Tl-201 activity was quantitatively measured in 22 myocardial segments. Regional LV function was assessed in corresponding segments by echocardiography. RESULTS: Agreement in the evaluation of regional perfusion status between tetrofosmin and Tl-201 imaging was observed in 78% of the 726 total segments, with a kappa value of 0.61. In segments with normal function at echocardiography (n = 436), no difference between Tl-201 and tetrofosmin uptake was observed. In hypokinetic segments (n = 138), exercise tetrofosmin uptake was lower (P < .01) as compared with exercise Tl-201 activity, whereas no difference was observed between tetrofosmin uptake at rest as compared with Tl-201 activity on redistribution and reinjection images. In segments with severe functional impairment (akinetic or dyskinetic, n = 152), tetrofosmin uptake on exercise images was reduced (P < .01) as compared with exercise Tl-201 activity; furthermore, tetrofosmin uptake at rest was lower (P < .01) as compared with Tl 201 activity on both redistribution and reinjection images. In these segments, concordance in the detection of myocardial viability between tetrofosmin and Tl 201 imaging was observed in 138 (91%) of the 152 segments, with a kappa value of 0.77. CONCLUSIONS: In patients with chronic coronary artery disease and LV dysfunction quantitative exercise-rest tetrofosmin and Tl-201 reinjection SPECT provide similar information in the assessment of perfusion status and in the detection of myocardial viability. PMID- 10385183 TI - Comparison of Tc-99m sestamibi and Tl-201 gated perfusion SPECT. AB - BACKGROUND: To determine the interpretability of gated thallium-201 perfusion SPECT compared with that performed by use of technetium-99m sestamibi (MIBI), 33 patients with prior myocardial infarction were studied. Patients received 22 to 30 mCi (814 to 1110 MBq) MIBI at peak stress, and a 15-minute gated SPECT acquisition was begun 30 to 40 minutes thereafter. On a subsequent day gated Tl 201 SPECT was acquired for 15 minutes, 4 hours after a resting 3.5 mCi (130 MBq) injection. SPECT was performed over a 180-degree arc by use of a 90-degree angled 2-detector camera. RESULTS: Gated studies were interpreted independently by 4 experienced physicians. Study quality was graded (0 = uninterpretable to 4 = excellent). Wall motion (0 = normal to 2 = akinetic/dyskinetic) and wall thickening (0 = normal to 2 = absent) were graded for each of 10 segments viewed in orthogonal planes. Left ventricular ejection fraction (LVEF) was calculated by use of software thus far validated only for MIBI. The average count density of mid-ventricular end-diastolic short axis tomograms with sestamibi was 3.47 times greater than with thallium. Mean study quality was 3.4 for MIBI and 1.8 for thallium (P < 10(-6)). No gated MIBI SPECTs, but 2 gated thallium studies (6%) were judged uninterpretable. Among interpretable scans, interobserver agreement (Kendall statistic) in assessing wall motion was 0.73 for MIBI and 0.66 for thallium (P = .01). For assessing wall thickening, the Kendall statistic was 0.73 for MIBI and 0.69 for thallium (P = .05). Correlation (r) of LVEFs was 0.91, SEE = 6.4. CONCLUSIONS: We conclude that gated thallium SPECT is inferior to MIBI because of much poorer image quality and somewhat poorer interobserver agreement among experienced physicians. However, LVEF can be determined reliably from gated thallium SPECT. PMID- 10385184 TI - Segmentation of gated Tl-SPECT images and computation of ejection fraction: a different approach. AB - BACKGROUND: We describe a set of image processing algorithms and mathematical models that can be advantageously used in schemes for the segmentation of thallium-201-single photon emission computed tomography (SPECT) images and for computation of left ventricular ejection fraction (EF). METHODS: The system consists of two independent blocs for image segmentation and computation of function. The former is based on a multiresolution elliptical coordinate transformation and dynamic contour tracking. Computation of EF is formulated on the basis of both the endocardial and epicardial contours, and we compare this formulation with that using only the endocardial border for images with low signal-to-noise ratios. The accuracy of border detection was validated against manual border tracing on FDG-PET images, simulated Tl-201-SPECT images where the true underlying borders were known, and actual Tl-201-SPECT images. Finally, we compared EFs computed for FDG-PET, technetium-99m-SPECT and Tl-201-SPECT with those obtained from planar gated blood pool imaging. RESULTS: The automatically obtained results always were within the manual uncertainty range. Agreement between myocardial volumes from positron emission tomography and automatically obtained values from the simulated Tl-201-SPECT images was excellent (r = 0.95, n = 32). Agreement between EFs from planar gated blood pool imaging and the other image modalities was good (FDG-PET: y = 5.89 + 1.21x, r = 0.92, see = 6.24, n = 19, Tc-99m-SPECT: y = -3.86 + 1.06x, r = 0.88, see = 7.78, n = 9, Tl-201-SPECT: y = 17.8 + 0.81x, r = 0.77, see = 7.44, n = 26). For noisy input data the combined use of information from epicardial and endocardial contours gives more accurate EF values than the traditional formula on the basis of the endocardial contour only. CONCLUSIONS: Alternate approaches for segmentation and computation of function have been presented and validated. They might also be advantageously incorporated into other existing techniques. PMID- 10385186 TI - HL-91-technetium-99m: a new marker of viability in ischemic myocardium. AB - BACKGROUND: Technetium 99m-HL91 is a new hypoxia imaging agent that demonstrates increased uptake in ischemic, viable myocardium. This study was performed to determine whether HL91 is taken up by nonviable myocardium. METHODS: Twenty-three Krebs-Henseleit buffer-perfused, isolated rat hearts were studied. Tc-99m-HL91 300 microCi was infused over 10 minutes, followed by a 60-minute clearance. Myocardial activity was monitored by use of an NaI crystal. Four groups were studied: control (flow = 12 mL/min, n = 7), low flow (flow = 1 mL/min, n = 6), no flow/reflow (60 minutes no flow/60 minutes reflow before Tc-99m-HL91 infusion, flow = 12 mL/min, n = 5), and cyanide-treated (before Tc-99m-HL91 infusion, flow = 12 mL/min, n = 5). Injury was assessed by creatine kinase, transmission electron microscopy, and triphenyltetrazolium chloride. RESULTS: Control (no injury) and cyanide-treated (severe injury) hearts demonstrated low uptake (6.3+/ 0.5 mean+/-SEM and 5.7+/-1.2 microCi, respectively) and low 60-minute retention (13.8%+/-2.2% and 13.7%+/-3.9%, respectively). Low-flow hearts (minimal injury) demonstrated markedly increased uptake (43.5+/-2.8 microCi, P < .01) and increased 60-minute retention (33.2%+/-2.9%, P < .01) compared with control. No flow/reflow hearts (moderate injury) demonstrated intermediate uptake (8.7+/-0.5 microCi, P < .05 to control), although retention was not significantly different (18.9%+/-3.5%, P = ns). Severely and rapidly injured myocardium demonstrated Tc 99m-HL91 peak uptake and retention indistinguishable from normal. Moderately injured myocardium demonstrated uptake intermediate between severely injured and low-flow-induced ischemic, viable myocardium. CONCLUSION: Thus Tc-99m-HL91 is not taken up or retained in nonviable and irreversibly injured myocardium. PMID- 10385185 TI - Establishing an approach for patients with recent coronary occlusion: identification of viable myocardium. AB - BACKGROUND: Revascularization of occluded coronary arteries after myocardial infarction (MI) may restore flow to viable myocardium and improve ventricular function. The aim of this pilot study was to determine the potential utility of thallium-201 viability imaging for the prediction of recovery of regional ventricular function in patients undergoing revascularization of total or subtotal occlusion of infarct-related arteries (TIMI 0-2 flow) during the convalescent period after MI. METHODS: Twenty-three patients were identified < 6 weeks after MI and underwent Tl-201 viability imaging (rest imaging, n = 16; stress/reinjection imaging, n = 7) and radionuclide angiography. Patients were revascularized with percutaneous transluminal coronary artery in 10, stent in 10, and bypass in 3. Follow-up radionuclide angiography at 3 months was used to assess recovery of regional wall motion. RESULTS: Among 41 abnormal wall motion segments in the infarct territories, the sensitivity, specificity, and accuracy for Tl-201 imaging in the prediction of recovery of regional function were 89% (25/28), 54% (7/13), and 78% (32/41), respectively. When 8 segments supplied by vessels with restenosis to >70% were excluded, specificity improved to 70%. Wall motion scores improved in those with adequate revascularization (1.6+/-1.4 vs 2.7+/-1.6; P < .001) but not in those with restenosis or occlusion (1.8+/-1.0 vs 2.0+/-1.6; P = NS). CONCLUSIONS: In patients with an occluded artery after MI, Tl 201 viability imaging can detect recoverable myocardium with reasonable accuracy and may help select which patients will most benefit from revascularization. PMID- 10385187 TI - Complementary roles of antibody affinity and specificity for in vivo diagnostic cardiovascular targeting: how specific is antimyosin for irreversible myocardial damage? AB - BACKGROUND: Identification of irreversible myocyte injury with antimyosin antibody imaging depends on both antibody specificity and affinity. To characterize the role of antibody affinity, we performed studies in dogs with acute coronary occlusion followed by reperfusion using 3 monoclonal antimyosin antibodies with different affinities. METHODS AND RESULTS: Dogs with experimental reperfused acute myocardial infarction were injected with 2 high-affinity radiolabeled monoclonal antimyosin Fab fragments (R11D10 and 2G42D7), 1 low affinity antimyosin Fab (3H31E6), and a nonspecific Fab. The left lateral gamma images at 5 H were used to assess the infarct (I) to blood (B) region of interest (ROI) count density ratios by computer planimetry. All infarcts were confirmed by in vivo imaging with 201Tl for perfusion defects as well as by postmortem histochemical staining. The mean I/B ROI (+/-standard deviation [SD]) for R11D10 (1.701+/-0.376) was not significantly different from that of 2G42D7 (1.501+/ 0.267, P = NS), but both were significantly greater than that of 3H31E6 Fab (0.85+/-0.12, P = .0001 and .0012, respectively). The I/B ROI of 3H31E6 Fab was similar to that of nonspecific Fab (0.75 to 0.77 range). Radiolabeled R11D10 and 2G42D7 were unequivocally positive by gamma imaging in all infarcts by 5 H. No infarcts were visualized with 3H31E6 or nonspecific Fab. CONCLUSIONS: The low affinity antibody, despite its specificity for cardiac myosin, cannot be used to image the infarct zone. Therefore immunoscintigraphic diagnosis of irreversible myocardial injury with radiolabeled antimyosin Fab is doubly specific because in vivo visualization required both specificity and high enough affinity of the antibody. PMID- 10385188 TI - Assessment of the severity of coronary artery stenosis by the ratio of the regional washout rate determined by adenosine triphosphate stress Tl-201 SPECT. AB - BACKGROUND: Adenosine triphosphate stress thallium-201 single-photon emission computed tomography (ATP SPECT) is useful for diagnosis of coronary artery disease, but its usefulness for evaluating the severity of coronary artery stenosis has not been established. METHODS AND RESULTS: We performed region-of interest analysis of short-axis images obtained by ATP SPECT in 31 patients with single-vessel disease (>50% stenosis of the luminal diameter). We selected the lowest and highest washout rates (WR) among the anterior, lateral, and inferior WRs and calculated the ratio of the lowest WR to the highest WR (WR ratio = 0.925+/-0.027 in 14 control subjects). ATP SPECT showed positive results in 29 (94%) of 31 patients. The severity of coronary artery stenosis was inversely correlated with the WR ratio (r = -0.703, P < .0001). The sensitivity and specificity of a WR ratio < or = 0.660 for the diagnosis of severe coronary stenosis (> or =80% stenosis) were 83% and 80%, respectively. CONCLUSIONS: Results suggest that ATP SPECT may be useful for assessment of the severity of coronary artery stenosis in patients with single-vessel disease. PMID- 10385190 TI - Imaging with radiolabeled antisense oligonucleotides for the detection of intracellular messenger RNA and cardiovascular disease. PMID- 10385189 TI - Ultra-high-resolution imaging of small animals: implications for preclinical and research studies. AB - Recent developments in the use of pinhole SPECT and dedicated PET for UHR small animal imaging have identified the technology that can be used to provide images with spatial resolution of the order of 1 to 3 mm. In SPECT imaging, rotating camera pinhole SPECT has provided the means to use existing equipment to achieve UHR images. It has the disadvantages of low sensitivity and requires special image software to reconstruct tomographic slices. However, with minimal additional cost to an imaging laboratory, pinhole SPECT can provide a quantitatively accurate imaging technique for small-animal studies. New detector technology offers considerable promise; however, more studies are required before any one system can be singled out as offering major advantages over the pinhole SPECT method for general purpose small-animal SPECT imaging. The search for the means to achieve better sensitivity with UHR continues. In PET imaging, with few exceptions, the general trend has been to design systems dedicated to small animal imaging to achieve UHR images with satisfactory sensitivity for quantitative UHR imaging. Several of the ring configuration, small-animal PET imaging systems provide good sensitivity and high spatial resolution. The cost of many of these systems, however, is relatively high, and investigators continue to explore different detector materials and imaging geometries to develop an instrument with acceptable levels of sensitivity with UHR imaging capability. We believe that both small-animal SPECT and PET imaging techniques now offer practical UHR imaging methods for quantitative small-animal imaging. As these tools are implemented in the investigation of new radiopharmaceuticals, we expect the utility of in vivo small animal assays will support further research in optimizing this technology. PMID- 10385191 TI - Heart medications: indications and interactions in stress testing. PMID- 10385193 TI - Blueprint of the Accreditation program of the Intersocietal Commission for the Accreditation of Nuclear Medicine Laboratories. PMID- 10385194 TI - Identification of severe right ventricular dysfunction and pressure overload by stress radionuclide myocardial perfusion SPECT imaging with gating. PMID- 10385192 TI - The challenge of quantifying defect size and severity: reality versus algorithm. PMID- 10385195 TI - Nuclear cardiology in the literature. PMID- 10385197 TI - Prophylaxis of local vascular graft infection with levofloxacin incorporated into albumin-sealed dacron graft (LVFX-ALB Graft). AB - An animal model was used to assess the efficacy of levofloxacin (LVFX) incorporated into albumin (ALB)-sealed Dacron (LVFX-ALB) graft for the prevention of vascular graft infections caused by Staphylococcus aureus. Under general anesthetic, an interposition graft was placed into dog carotid artery. On completion of the operation, 0.1 ml of normal saline containing 10(7) colony forming units (CFU) of a slime-producing S. aureus was inoculated directly onto the graft. After 1 day, the samples were sterilely harvested. The antibacterial activity of LVFX into the LVFX-ALB graft was evaluated by colony counting in bacterial cultures and by the fluorescent antibody method staining bacteria adhesion to the grafts. LVFX-ALB grafts had a lower infection rate than the control grafts (1/4, 10(2) CFU vs 4/4, 1.50 x 10(5)+/-1.38 x 10(5)CFU (mean+/ SE)). In an immunostaining study, LVFX-ALB grafts had small fluorescent areas showing S. aureus adhesion, while fluorescence was observed over the entire surface of the control grafts. Therefore, LVFX-ALB presumably had a bactericidal action and adhesive prevention against inoculated S. aureus. LVFX-ALB may be useful in preventing graft infections during and immediately after vascular reconstruction. PMID- 10385196 TI - Antibacterial activities of new synthetic divalent cation chelators. AB - A series of new synthetic ligand compounds which chelate divalent cations was examined for the antibacterial activities of the compounds. Only 2 of 14 synthetic chelators, 9-trans-anthryl-1, 4, 8, 11-tetraaza-tetradecane (No. 6) and bis(2-pyridyl)methylamine (No. 13) showed antibacterial activities, whereas none of the diamines, hydrophilic triamines nor tetramines showed antibacterial activities. Chelators No. 6 and No. 13 inhibited the growth of both Gram-negative and -positive bacteria at doses of 25-200 microg/ml, comparable to those of common antibiotics such as polymixin B, fosfomycin and macrolides. Ethylenediaminetetraacetate (EDTA) potentiated these antibacterial activities, whereas an inhibitory effect of Mg2+ on the MICs of these chelators was observed. Moreover, these chelators enhanced the leakage of periplasmic beta-lactamase. It is therefore suggested that chelators No. 6 and No. 13 disrupt both the membranes and cytoplasmic functions of bacteria, resulting in cell death. PMID- 10385198 TI - Role of superoxide anion in the fungicidal activity of murine peritoneal exudate macrophages against Penicillium marneffei. AB - Penicillium marneffei is an important opportunistic fungal pathogen. The mechanisms of host defense against P. marneffei are not fully understood. In the present study, we, for the first time, investigated the role of superoxide anion (O2-) in the killing of two forms of P. marneffei, yeast cells and conidia, and the role of this killing mediator in the fungicidal activity of IFN-gamma stimulated murine peritoneal macrophages. P. marneffei yeast cells were susceptible to the killing effect of activated macrophages and chemically generated O2, while conidia were not. These results suggested that O2- played some role in the fungicidal activity of macrophages. However, an oxygen radical scavenger, superoxide dismutase (SOD), did not suppress, but rather enhanced the fungicidal activity of IFN-gamma-stimulated macrophages against P. marneffei yeast cells. This inconsistency was explained by the release of insufficient concentrations of O2- by activated macrophages as compared with the amount of O2- necessary for the killing of yeast cells, which was predicted in a chemical generating system. On the other hand, SOD enhanced the production of nitric oxide (NO) by IFN-gamma-activated macrophages, and their increased fungicidal activity was significantly inhibited by N(G)-monomethyl-L-arginine (L-NMMA), a competitive inhibitor of NO synthase. Our results suggested that O2- does not function as the killing mediator of macrophages against P. marneffei, but rather plays an important role in the regulation of the NO-mediated killing system by suppressing NO production. PMID- 10385199 TI - In vitro assessment of a chemically synthesized Shiga toxin receptor analog attached to chromosorb P (Synsorb Pk) as a specific absorbing agent of Shiga toxin 1 and 2. AB - A synthetic analog of Shiga toxin (Stx) receptor (Synsorb Pk) was quantitatively assessed to determine whether it can protect human renal adenocarcinoma cells (ACHN cells) from the cytotoxicity of Stx1 and Stx2 by coincubation experiments. Coincubation of 100 and 20 ng of Stxl and Stx2 with 50 mg of Synsorb Pk for 1 hr at 37 C in 1 ml of Eagle's Minimum Essential Medium supplemented with 1% (v/v) non-essential amino acid and 10% (v/v) fetal calf serum protected 50% of the cells from the cytotoxic effect. Chromosorb P, an inert matrix control, did not absorb the Stxs at all. Heat-treatment (boiled for 10 min) to Synsorb Pk caused a 50% decrease in Stx2-binding activity, but did not effect the Stx1 binding. Further, Stxs bound to Synsorb Pk could be demonstrated. When 20 mg of Synsorb Pk was coincubated for 30 min at 37 C in 1 ml of phosphate-buffered saline with 1 and 10 ng or more of Stx1 or Stx2, respectively, the toxins could be detected on the surface when the bound toxins on Synsorb Pk were used as the solid phase in enzyme immunoassay. The amount of 100 ng/ml of both Stxl and Stx2 appeared to saturate 20 mg/ml of Synsorb Pk after coincubating for 30 min at 37 C. While assessing the Stxs' binding activity to Synsorb Pk, it was demonstrated that Stxl had a higher affinity to Pk trisaccharide than Stx2. These observations provide useful information on the effectiveness of Synsorb Pk to trap and eliminate free Stxs produced in the gut of patients infected by Stx-producing Escherichia coli, and to prevent the progression of hemorrhagic colitis to hemolytic uremic syndrome. PMID- 10385200 TI - Proposal of Sphingomonas suberifaciens (van Bruggen, Jochimsen and Brown 1990) comb. nov., sphingomonas natatoria (Sly 1985) comb. nov., Sphingomonas ursincola (Yurkov et al. 1997) comb. nov., and emendation of the genus Sphingomonas. AB - Based on the results of a phylogenetic analysis of 16S rRNA and the presence of sphingoglycolipid in cellular lipids of the type strains, transfer of "Rhizomonas" suberifaciens, Blastomonas natatoria and Erythromonas ursincola to the genus Sphingomonas as Sphingomonas suberifaciens (van Bruggen et al 1990) comb. nov., Sphingomonas natatoria (Sly 1985) comb. nov., and Sphingomonas ursincola (Yurkov et al 1997) comb. nov. are herein proposed together with the emendation of genus Sphingomonas. The type strain of S. suberifaciens is van Bruggen Cal=ATCC 49382=NCPPB 3629=IFO 15211=JCM 8521, that of S. natatoria is ATCC 35951 =DSM 3183=NCIMB 12085=JCM10396, and that of S. ursincola is DSM 9006= KR-99. PMID- 10385201 TI - TTG as the initiation codon of Salmonella slyA, a gene required for survival within macrophages. AB - The slyA gene, which has been implicated in the virulence of Salmonella serovar Typhimurium and its survival in macrophages, is widely distributed among different Salmonella serovars. In this study, we cloned and sequenced the translational initiation region of the slyA gene from nine different serovars and found sequence differences in the previously proposed ATG initiation codon but not in a TTG triplet, another putative initiation codon in the slyA gene. Therefore, we determined the actual translational initiation site of the slyA gene by analyzing slyA genes with defined mutation in either the ATG or TTG sequences in an in vitro translation assay and a quantitative hemolytic assay in Escherichia coli. The replacement of TTG by TTC in the slyA gene significantly reduced both the amount of protein synthesized and the hemolytic activity of a transformed strain of E. coli, while replacement of ATG by ATC had no effect in these assays. In addition, the amino acid sequence analysis of the His-tagged SlyA protein showed that it was identical with the amino acid sequence deduced from the 5' end of the slyA gene with a TTG initiation codon. Our results suggest that TTG serves as the translational initiation codon for the slyA gene of Salmonella. PMID- 10385202 TI - PCR-RFLP analysis of cytomegalovirus infections associated with bone marrow transplantation in Japanese children. AB - In order to investigate the longitudinal molecular epidemiology of cytomegalovirus (CMV) infections associated with bone marrow transplantation (BMT) in Japanese children, we analyzed 36 CMV strains from 11 cases. Three regions (DNA polymerase, glycoprotein H, and immediate-early regions) of CMV DNA were amplified by polymerase chain reaction (PCR), and amplified products were each digested with two restriction enzymes, followed by electrophoresis. These restriction fragment length polymorphism (RFLP) analyses allowed the differentiation of 36 strains into 13 genotypes. Each patient excreted his or her own CMV with distinct genotype over the study period of up to one year. CMVs of two different genotypes were recovered during a one-month study from one recipient, who received a peripheral blood stem cell transplantation. Although the majority of patients and donors were CMV-seropositive before BMT, multiple CMV infections might not be common and the reactivation of latently infected CMV might be prominent in Japanese children receiving transplants. PMID- 10385203 TI - Overproduction of gamma interferon in B/Jas inbred rabbits with herpes simplex virus encephalitis. AB - Inbred rabbits of the B/Jas strain are highly susceptible to herpes simplex virus type-1 (HSV-1) encephalitis, developing seizures of encephalitis after intravenous injection of the KOS strain of the virus. Anti-viral interferon activity became detectable in the serum just prior to or at the onset of seizures, its level being lower in the serum than in the cerebrospinal fluid. The activity was of gamma interferon, as suggested by the acid instability and the inability of Mx protein induction. An immunohistochemical analysis of the brain tissues of encephalitic rabbits showed that MHC class I antigen was expressed on the microglia cells of inflamed lesions but not on these cells in uninflamed areas. These findings were discussed in correlation with the pathogenesis of herpetic encephalitis in the inbred rabbits. PMID- 10385204 TI - Genetic variation in the 5' end and NS5B regions of classical swine fever virus genome among Japanese isolates. AB - Sixteen clinical strains of classical swine fever virus (CSFV) isolated in Japan were subjected to analyses of nucleotide sequence variations in the 5' end and NS5B regions of the genome. These isolates were divided into three genovars, CSFV 1, CSFV-2 and CSFV-3, based on palindromic nucleotide substitutions at the three variable loci in the 5' untranslated region (UTR). Phylogenetic trees constructed from nucleotide sequences in the 5'-UTR and NS5B gene indicated that the CSFV strains were divided into three clusters, I, II and III. CSFV strains included in clusters I, II and III were identical to those in the CSFV-1, CSFV-2 and CSFV-3 genovars, respectively. PMID- 10385206 TI - A filamentous phage of Vibrio parahaemolyticus O3:K6 isolated in Laos. AB - A filamentous phage, 'lvpf5,' of Vibrio parahaemolyticus O3:K6 strain LVP5 was isolated and characterized. The host range was not restricted to serotype O3:K6, but 7 of 99 V. parahaemolyticus strains with a variety of serotypes were susceptible to the phage. The phage was inactivated by heating at 80 C for 10 min and by treating with chloroform. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the phage exhibited a 3.8 kDa protein. The amino-terminal amino acid sequence of the coat protein was determined as AEGGAADPFEAIDLLGVATL. The phage genome consisted of a single-stranded DNA molecule. The activity of the phages was inhibited by anti-Na2 pili antibody. PMID- 10385205 TI - Molecular epidemiological study of a mass outbreak caused by enteropathogenic Escherichia coli O157:H45. AB - We made a molecular analysis of O157:H45 Escherichia coli isolated from a mass outbreak that occurred in Obihiro City. Using DNA analysis, we confirmed this infection case as a mass outbreak. Although the isolates expressed O157 antigen, they did not produce Vero toxin. We concluded they were enteropathogenic E. coli (EPEC) because they had a bfp gene and an EAF plasmid, and further they exhibited local adherence to HEp-2 cells. We believe this is the first report of a mass outbreak by O157 EPEC, and we suggest that PCR using eae- and bfp-specific primers and HEp-2 adherence assay are useful to identify EPEC. PMID- 10385207 TI - Preparation, characterization and biological evaluation of copper(II) and zinc(II) complexes with cephalexin. AB - Copper(II) and zinc(II) complexes of cephalexin have been prepared and characterized by microanalysis and by thermogravimetric, magnetic and spectroscopic analysis. The complexes were found to be five-coordinate, monohydrate, and ML2 type. The electron paramagnetic resonance spectral lines revealed rhombic distortion from axial symmetry, with g(parallel) > g(perpendicular) > g(e), in the elongated-tetragonal copper(II) complex. The geometry of the zinc(II) complex seems to be square-pyramidal. On complexation with copper and zinc the antimicrobial activity of cephalexin improved significantly. The copper complex was found to be active against kaolin paw oedema whereas the parent drug was inactive. These results suggest that the metallic elements should be seriously considered during drug design, and that complexes already reported should be subjected to clinical evaluation. Their use could provide an easy way of improving the activity and reducing the toxicity of drug substances. PMID- 10385208 TI - Single- and repeated-dose local toxicity in the nasal cavity of rabbits after intranasal administration of different glycols for formulations containing benzodiazepines. AB - To furnish a systemic effect after intranasal administration, a formulation must contain the therapeutic dose in no more than 150 L, the maximum volume that can be applied as a single administration in one nostril in man. The objectives of these studies were to examine the local toxicity of formulations containing benzodiazepines and to document the effects to support clinical trials in man. After stability, pharmacological and pharmacokinetic studies of several benzodiazepine formulations, we studied nasal toxicity after single and repeated administration to rabbits of poly(ethylene glycol) 200, tetra(ethylene glycol), glycofurolum and mixtures of these vehicles both with and without benzodiazepines. Single-dose studies with examinations 5 or 10min after application were undertaken with poly(ethylene glycol), tetra(ethylene glycol), glycofurolum and tetra(ethylene glycol)-glycofurolum in the ratio 95:5; the reactions were similar to that after physiological saline. A 14-day repeated-dose study was conducted with diazepam, lorazepam and flunitrazepam formulations in poly(ethylene glycol), and flunitrazepam in poly(ethylene glycol)-glycofurolum in the ratio 70:30; the two vehicles without any benzodiazepine were also examined. Microscopic study revealed mild changes only in the treated groups. A final four week study was conducted with repeated administration of clonazepam formulated in tetra(ethylene glycol)-glycofurolum in the ratio 95:5; microscopy revealed mild changes after three 150-microL doses daily, but no abnormalities after one or three 100-microL doses daily. It was concluded that these three solvents individually or as mixtures resulted in only mild local toxicity and might be acceptable as vehicles in nasal preparations of benzodiazepines and other non irritating drugs for short-term use in man. PMID- 10385209 TI - Stability study of human serum albumin pharmaceutical preparations. AB - The influence of temperature on the stability of human serum albumin (HSA) pharmaceutical preparations has been studied by size-exclusion high-performance liquid chromatography with multi-angle laser-light-scattering detection and by particle-size analysis. The behaviour of HSA in two pharmaceutical preparations stored at different temperatures (40, 55 and 70 degrees C) followed the same pattern--an increase in the relative percentage of dimer (MW 132 000 Da) and aggregate (MW > 200 000 Da), and then a decrease in the concentration of all species and, finally, sudden protein coagulation. These results suggest a time- and temperature-dependent process. At 70 degrees C, monomer only was detected for both preparations; the amount remaining was 83 and 72% for formulations A and B, respectively. Analysis of size-distribution curves also seems to confirm these results. Initially, three distributions were observed with length-volume mean diameters (d1,v) of 1.67, 10.6 and 57 microm. After 80 days at 55 degrees C, only two distributions were observed, with d1,v of 3.07 and 76 microm. An additional study using pure HSA at different concentrations (0.3, 2.5, 5 and 10% w/v) and storage at 75 degrees C was performed to determine the influence of the concentration of auxiliary substances and of the HSA. Only when the HSA concentration was 0.3% w/v did the remaining fraction of HSA fit a Prout Thompkins nucleation model. Initially three distributions with mean sizes of 2, 20 and 40 microm were observed whereas at the end of the assay only one distribution, mean size 129 microm, was seen. The methodology used enabled us to separate the HSA degradation products and to determine the absolute molecular weight of albumin monomer and dimer. It is possible to conclude that the degradation mechanism for the formulations studied is complex, and that it is possible to fit the degradation data to Prout-Thompkins kinetics only when the concentration of HSA is low enough (0.3% w/v). PMID- 10385210 TI - Pharmacokinetics of ticlopidine in the rabbit. AB - There is no information about the pharmacokinetics of ticlopidine in rabbits. Such information is valuable in designing appropriate dosing regimens for experimental studies of the drug with ultimate applications in man. The disposition kinetics of ticlopidine at three dose levels were evaluated in three groups of six rabbits which received 10, 50 or 100 mgkg(-1) drug once daily via the oral-gastric route. Blood samples were collected at predetermined times after the third dose. Plasma concentrations of the unchanged drug were determined by a validated liquid chromatography-mass spectrometry method with a limit of detection of 5 microg L(-1). There was a disproportionate increase in the mean maximum plasma concentration (Cmax) and the area under the plasma drug concentration-time curve (AUC) for the 10 and 50 mgkg(-1) doses. The apparent terminal half-life (t1/2beta), apparent volume of distribution (Vdbeta/F), and total plasma clearance (CLp/F) of the drug were all dose-dependent. For example, t1/1beta for the 10, 50 and 100 mgkg(-1) doses were 1.04+/-0.10, 4+/-24+/-1.92 and 12.80+/-6.35 h, respectively, whereas the Vdbeta/F values for the corresponding doses were 214 31, 475 221 and 998+/-420 Lkg(-1), respectively. These results show that the 100-mgkg(-1) dose produces plasma ticlopidine concentrations similar to those found in man after administration of 250 mg of the drug. It is suggested that 100 mg kg(-1) might be the appropriate dose of ticlopidine for use in rabbit experimental studies with ultimate application to man. PMID- 10385211 TI - Influence of glycerol-induced acute renal failure on the pharmacokinetics of cyclosporin in rats. AB - Although it is widely believed that renal dysfunction has no effect on the pharmacokinetics of cyclosporin, many clinical reports suggest that renal dysfunction after renal transplantation is closely related to the pharmacokinetics of cyclosporin. To clarify the relationship between renal dysfunction and the pharmacokinetics of cyclosporin, we examined the influence of acute renal failure (ARF) on its pharmacokinetics in glycerol-induced ARF rats. The values of indicators of renal function (serum creatinine, blood urea nitrogen), but not those of indicators of hepatic function, were significantly increased in ARF rats that received glycerol compared with values for control rats. The area under the blood cyclosporin concentration-time curve after oral administration (AUCpo) were 4.976+/-0.847 mghL(-1) for ARF rats and 9.684+/-1.100 mghL(-1) for control rats; AUCpo in ARF was significantly reduced in a manner dependent on renal function. The oral clearance of cyclosporin in ARF and control rats was 1.172+/-0.207 and 0.544+/-0.062Lh(-1) kg(-1), respectively, whereas total body clearance in ARF and control rats was 0.151+/-0.008 and 0.183+/ 0.010Lh(-1)kg(-1), respectively. The relative bioavailability of cyclosporin in ARF and control rats was 0.118 and 0.336, respectively. In an in-vitro study using everted sac and liver-slice methods, the apparent first-order rate constants for cyclosporin uptake (k(uptake)) and metabolism (k(metab)) in gut tissues were reduced, whereas k(uptake) and k(metab) in liver were increased. Gastric emptying, measured by use of paracetamol, was significantly reduced in ARF rats. These results suggest that glycerol-induced ARF results in several changes in the pharmacokinetics of cyclosporin in rats. From these results, we conclude that reduction of the absorbed fraction of cyclosporin strongly contributes to the decrease in AUCpo in the presence of ARF. PMID- 10385212 TI - In-vivo and in-vitro metabolic clearance of midazolam, a cytochrome P450 3A substrate, by the liver under normal and increased enzyme activity in rats. AB - The metabolic clearance of midazolam, a cytochrome P450 (CYP) 3A substrate, by the liver under normal and increased enzyme activity in rats was determined in vivo and in-vitro to elucidate the reproducibility of the in-vivo hepatic extraction ratio of midazolam from the in-vitro study. The hepatic enzyme activity was modified by pretreating rats with a CYP inducer such as dexamethasone and clotrimazole. The in-vivo hepatic extraction ratio (ERh,obs) of midazolam under a steady-state plasma concentration (approx. 3 nmolmL(-1)) in untreated (control) rats was 0.864. This value increased to 0.984 in dexamethasone-pretreated rats and to 0.964 in clotrimazole-pretreated rats. The in-vitro hepatic intrinsic clearance (CL(int,in-vitro)), expressed as mLmin(-1) (mg microsomal protein)(-1), of midazolam was estimated as Vmax (Km)(-1) by in vitro metabolism studies using liver microsomes. The CL(int,in-vitro) value was converted to the CL(int,cal) value, expressed as mLmin(-1)kg(-1), by considering the microsomal protein content (g liver)(-1) and the microsomal protein content (g liver)(-1)kg(-1). The estimated CL(int,cal) value was then converted to the ERh value (ER(h,cal)) according to the well-stirred, the parallel-tube and the dispersion models. The ERh(h,cal) values obtained by the parallel-tube model were in good agreement with corresponding in-vivo ERh(h,obs) values. In conclusion, it was demonstrated that high hepatic clearances of midazolam under normal and increased CYP3A activity were reasonably predicted from in-vitro metabolism studies using liver microsomes. PMID- 10385213 TI - Aldehyde oxidase-catalysed oxidation of methotrexate in the liver of guinea-pig, rabbit and man. AB - Although 7-hydroxymethotrexate is a major metabolite of methotrexate during high dose therapy, negligible methotrexate-oxidizing activity has been found in-vitro in the liver in man. The goals of this study were to determine the role of aldehyde oxidase in the metabolism of methotrexate to 7-hydroxymethotrexate in the liver and to study the effects of inhibitors and other substrates on the metabolism of methotrexate. Methotrexate, (+/-)-methotrexate and (-)-methotrexate were incubated with partially purified aldehyde oxidase from the liver of rabbit, guinea-pig and man and the products analysed by HPLC. Rabbit liver aldehyde oxidase was used for purposes of comparison. In-vitro aldehyde oxidase from the liver of man catalyses the oxidation of methotrexate to 7-hydroxymethotrexate, but the turnover is low. However, formation of 7-hydroxy-methotrexate from all forms of methotrexate by the liver in guinea-pig and man was significantly inhibited in the presence of 100 microM menadione and chlorpromazine, potent inhibitors of aldehyde oxidase. Allopurinol (100 microM) had a negligible inhibitory effect on liver aldehyde oxidase from guinea-pig and man. Allopurinol is a xanthine oxidase inhibitor. The production of 7-hydroxymethotrexate was enhanced in the presence of allopurinol. Although aldehyde oxidase is also responsible for some of this conversion, it is also possible that the closely related xanthine oxidase is responsible for the formation of 7 hydroxymethotrexate. By employing potent selective inhibitors of aldehyde oxidase, menadione and chlorpromazine, we have demonstrated for the first time that liver aldehyde oxidase from man is minimally involved in methotrexate oxidation. PMID- 10385214 TI - Halofantrine metabolism in microsomes in man: major role of CYP 3A4 and CYP 3A5. AB - We have clarified the contribution of the different enzymes involved in the N debutylation of halofantrine in liver microsomes in man. The effect of ketoconazole and cytochrome P450 (CYP) 3A substrates on halofantrine metabolism has also been studied. The antimalarial drug halofantrine is metabolized into one major metabolite, N-debutylhalofantrine. In microsomes from nine livers from man, N-debutylation of halofantrine was highly variable with apparent Michaelis-Menten constant V(max) and K(m) values of 215+/-172 pmol min(-1) mg(-1) and 48+/-26 micromol L(-1), respectively, (mean+/-standard deviation). Formation of N debutylhalofantrine was cytochrome P450 (CYP)-mediated. Studies using selective inhibitors of individual CYPs revealed the role of CYP 3As in the formation of N debutylhalofantrine. alpha-Naphthoflavone, a CYP 3A activator, increased metabolite formation. In microsomes from 12 livers from man the rate of N debutylation of halofantrine correlated strongly with CYP 3A4 relative levels (P = 0.002) and less strongly, but significantly, with CYP 2C8 levels (P = 0.025). To characterize CYP-mediated metabolism of halofantrine further, incubations were performed with yeast microsomes expressing specific CYP 3A4, CYP 3A5, CYP 2D6, CYP 2C8 and CYP 2C19 from man. The rate of formation of N-debutylhalofantrine was six- and twelvefold with CYP 3A4 than with CYP 3A5 and CYP 2C8, respectively. CYP 2D6 and CYP 2C19 did not mediate the N-debutylation of halofantrine, but, because in-vivo CYP 2C8 is present at lower concentrations than CYP 3A in the liver in man, the involvement of CYP 3As would be predominant. Diltiazem, erythromycin, nifedipine and cyclosporin (CYP 3A substrates) inhibited halofantrine metabolism. Similarly, ketoconazole inhibited, non-competitively, formation of N debutylhalofantrine with an inhibition constant, K(i), of 0.05 microM. The theoretical percentage inhibition of halofantrine metabolism in-vivo by ketoconazole was estimated to be 99%. These results indicate that both CYP 3A4 and CYP 3A5 metabolize halofantrine, with major involvement of CYP 3A4. In-vivo, the other CYPs have a minor role only. Moreover, strong inhibition, and consequently increased halofantrine cardiotoxicity, might occur with the association of ketoconazole or other CYP 3A4 substrates. PMID- 10385215 TI - Inhibition of aromatase (P450Arom) by some 1-(benzofuran-2-ylmethyl)imidazoles. AB - Studies of a series of 1-(benzofuran-2-ylmethyl)imidazoles, 1-5, previously proposed as potential agents for prostatic cancer by their inhibition of 17beta hydroxylase:17,20-lyase (P450 17), have been extended to their selectivity against placental microsomal aromatase (P450(Arom)) in man. The compounds were 3 7-fold more potent than aminoglutethimide and had some selectivity for P450 17 as expressed by the ratio (IC50 P450(Arom))/(IC50 P450) 17)/17.0 (2), 10.3 (3), 34.6 (4) and 42.0 (5), where IC50 is the concentration resulting in 50% inhibition. The lower potency of 1-5 towards P450(Arom) compared with the racemic alpha phenyl-substituted compounds (6, 80-1000 x aminoglutethimide) and some racemic alpha-methyl (8.5 and 12.2 x aminoglutethimide) and alpha-ethyl (12.1 and 32.9 x aminoglutethimide) analogues has been rationalized. This work selectively extends studies of the P450 17 inhibitor 5, a potential prostatic cancer agent, towards other cytochrome P450 enzymes in the steroidogenic pathway and provides a general method for determining the relative influence of chemical manipulation of a parent inhibitor towards two enzymes in the pathway using additional literature data. PMID- 10385216 TI - Protective effects of pseudoginsenoside-F11 on scopolamine-induced memory impairment in mice and rats. AB - This study assessed the effects of pseudoginsenoside-F11, a component of Panax quinquefolium L., on scopolamine-impaired memory performance in mice and rats. In the one-trial step-down and step-through passive avoidance tests, although pseudo ginsenoside-F11 used alone did not affect passive avoidance behaviour in naive mice, the latency of avoidance shortened by intraperitoneal scopolamine (2 mg kg( 1)) was prolonged after intragastric administration of pseudoginsenoside-F11 (2 or 4 mg kg(-1), for five days) in both test systems in mice. In the water-maze test, in mice, the time taken to locate the platform after administration of pseudoginsenoside-F11 was shorter than that after administration of scopolamine (1 mg kg(-1), i.p.). In the two-way active avoidance response test, the latency of avoidance was significantly shorter for the pseudoginsenoside-F11-(1.2 or 2.4 mg kg(-1), i.g. for five days) and scopolamine-treated group than for the group of rats given scopolamine only (2 mg kg(-1), i.p.). The percentage avoidance was also reduced after intraperitoneal injection of scopolamine, but was reversed by administration of pseudo-ginsenoside-F11. These results suggest that pseudoginsenoside-F11 antagonized the memory dysfunction induced by scopolamine. However, the mechanism of the memory facilitative action of pseudoginsenoside-F11 merits further elucidation. PMID- 10385217 TI - Effects of nicardipine on collar-induced intimal thickening and vascular reactivity in the rabbit. AB - The effects of nicardipine treatment on collar-induced intimal thickening and on accompanying reactivity changes in rabbit carotid artery have been investigated. Treatment for three weeks with subcutaneous nicardipine (20 mgkg(-1) per day) significantly inhibited the intimal thickening caused by perivascular application of a silicone rubber collar. Potassium chloride (KCl), phenylephrine and 5 hydroxytryptamine (5-HT) induced concentration-dependent contractions in both sham-operated and collared arteries. Collar-induced attenuation of maximum KCl-, phenylephrine- and 5-HT-induced contraction was not affected by nicardipine. Collaring caused the means of pD2 values (the negative logarithm of EC50 values, 50% effective concentration) of 5-HT and phenylephrine to increase and decrease, respectively. Nicardipine did not affect the altered sensitivity to these agonists. Neither collar implantation nor nicardipine treatment altered the pD2 values for acetylcholine- and nitroglycerine-induced relaxations. These results demonstrate that nicardipine inhibits collar-induced intimal thickening in rabbit carotid artery without affecting the accompanying changes in vascular reactivity, indicating a possible lack of association between the development of intimal thickening and altered reactivity. PMID- 10385218 TI - The action of 5-hydroxytryptamine on normal and cystic fibrosis mouse colon: effects on secretion and intracellular calcium. AB - The ability of mouse colon to generate a secretory response to stimulation by 5 hydroxytryptamine (5-HT) was investigated in intact colonic sheets mounted in Ussing chambers. A preparation of intact isolated crypts was used to determine whether 5-HT action was associated with an elevation of cytosolic calcium levels, measured using the calcium-sensitive fluorescent dye, fura-2. 5-HT increased the short-circuit current, an effect that was inhibited by 55% in the absence of chloride and by 83% in the presence of serosal frusemide, consistent with the stimulation of electrogenic chloride secretion. This was confirmed by the observation that colonic tissue from transgenic cystic fibrosis mice (n = 4) failed to respond to 5-HT, although wild-type tissues generated an increased short-circuit current of 52.4+/-1.1 microAcm(-2) (n = 9). The electrical response to 5-HT was calcium-dependent. 5-HT action was unaffected by tetrodotoxin and was not mimicked by the 5-HT3 agonist 1-phenylbiguanide, indicating that neural mechanisms are not involved. The cyclooxygenase inhibitor indomethacin, however, reduced the 5-HT-induced rise in short-circuit current by 73%, suggesting that prostaglandin production contributes to the response. Stimulation of crypts with acetylcholine elicited an increase in cytosolic calcium levels, but no such response was detected on application of 5-HT (10(-6) to 10(-4) M), suggesting that 5-HT does not directly modulate intracellular calcium in colonic crypt cells. It is concluded that mouse colon responds to 5-HT challenge with a stimulation of electrogenic chloride secretion and that this effect is mediated by indirect mechanisms that might involve immune elements within the colonic wall. PMID- 10385219 TI - The long-lasting effect of TU-199, a novel H+, K(+)-ATPase inhibitor, on gastric acid secretion in dogs. AB - We have used Heidenhain-pouch dogs to investigate the effects of (+/-)-5-methoxy 2-{[(4-methoxy-3,5-dimethylpyrid-2-yl)methyl]sulph inyl}-1H-imidazo[4,5 b]pyridine (TU-199), an imidazopyridine derivative, on gastric acid secretion stimulated by histamine, carbachol and tetragastrin. We have also investigated the duration of the antisecretory effect of TU-199 using a measurement of intragastric pH for 24 h in gastric fistula dogs whose gastric acid secretion was stimulated by histamine. Single oral administration of TU-199 (0.1, 0.2 and 0.4mgkg(-1)) dose-dependently suppressed gastric acid secretion stimulated by histamine infusion. Oral treatment with TU-199 (0.2, 0.4 and 0.8 mg kg(-1)) also dose-dependently inhibited acid secretion induced by carbachol and tetragastrin. The inhibitory effect of TU-199 on stimulated gastric acid secretion was more potent than that of omeprazole, a well-known H+,K(+)-ATPase inhibitor in dogs. Repeated oral treatment with TU-199 at a dose of 0.2 mg kg(-1) once a day for seven days markedly suppressed histamine-stimulated gastric acid secretion in dogs. This inhibitory effect of TU-199 reached a maximum level after three or four doses and was more pronounced than that of omeprazole or lansoprazole. In gastric fistula dogs, the duration of intragastric pH-elevation by administration of TU-199 (0.3 mg kg(-1)) was much longer than that of omeprazole (0.6mgkg(-1)) or lansoprazole (0.9mgkg(-1)). The IC50 values (doses resulting in 50% inhibition) of TU-199, omeprazole and lansoprazole with regard to H+,K(+)-ATPase activity in dog gastric mucosal microsomes were 8.6, 8.8 and 9.9 microM, respectively. These results indicate that TU-199 inhibits gastric acid secretion via suppression of a H+,K(+)-ATPase activity. Our findings also suggest that TU 199 might have potent and long-lasting effects on gastric acid secretion. PMID- 10385220 TI - Effects of interferons on cortisol production in bovine adrenal fasciculata cells stimulated by adrenocorticotropin. AB - The effects of interferons (IFNs) IFN-alpha, IFN-beta and IFN-gamma on the production of cortisol in bovine adrenal fasciculata cells have been investigated. Pretreatment of the fasciculata cells with recombinant interferon alpha-2b from man (over 300 units mL(-1)), but not with fibroblast IFN-beta or recombinant IFN-gamma from man, reduced the production of cortisol in cells stimulated with adrenocorticotropin (ACTH) (1 nM). IFN-alpha-2b inhibited ACTH induced cortisol production in a concentration- (300-15000 units mL(-1)) and time (2-24h) dependent manner. The inhibitory effect of IFN-alpha-2b on the production was abolished when the cells were simultaneously treated with anti-IFN alpha antibody, and it was reversible. IFN-alpha-2b also inhibited dibutyryl cyclic AMP-induced production of cortisol but not pregnenolone-induced production. The effect of IFN-alpha-2b was not influenced by increases in external ACTH and Ca2+ concentrations and IFN-alpha-2b did not affect the ACTH induced increase in cyclic AMP level in the cells. These results strongly suggest that IFN-alpha-2b reduces ACTH-induced production of cortisol in bovine adrenal fasciculata cells by affecting the early process of cortisol synthesis. The results also indicate that IFNs might not directly affect steroidogenesis in the adrenal cortex in-vivo, because of the requirement of high concentrations of IFN alpha-2b for inhibition, and because of the ineffectiveness of IFN-beta and IFN gamma. PMID- 10385221 TI - Effect of plasma-calcium-level-responsive oestradiol release from apatitic bone cement on bone mineral density in ovariectomized rats. AB - The effects of plasma calcium levels on oestradiol release from apatite bone cement and on the bone mineral density of ovariectomized rats have been investigated. Apatite cement was prepared from an equimolar mixture of tetracalcium phosphate, dicalcium phosphate dihydrate and 0.5% beta-oestradiol bulk powder. After subcutaneous implantation of the cement, oestradiol release in diseased rats (ovariectomized rats on a low-calcium diet) was significantly higher than in normal rats. The drug levels of recovery-model rats (ovariectomized, but on a high-calcium diet) were significantly lower than those of the diseased rats. Calcium levels in diseased rats remained low during drug release but the plasma calcium levels of the recovery-model rats increased. The areas under the plasma calcium concentration-time curves (Ca-AUCs) for the recovery-model rats were higher than those for the diseased-model rats. The plasma oestradiol concentration AUCs and the Ca-AUCs were linearly related. The body weight of the recovery-model rats increased after five days, but that of the diseased-model rats did not. The bone mass of the recovery-model rats was greater after the experiment than before. The relationship between the bone mineral density and Ca-AUC of the diseased rats suggested that bone mineral density increased with increasing Ca-AUC. The results suggest that the severity of osteoporosis in this animal model is reduced by implantation of the oestradiol loaded apatite cement. PMID- 10385223 TI - Preparation of PEG-coated surfaces and a study for their interaction with living cells. AB - Cell-biomaterial interaction is of great importance for the development of bioinert as well as of hybrid surfaces. This study represents our results of human fibroblast interaction with PEG-coated surfaces of differing length and structure (linear or branched) of the oxyethylene chain. We employed three PEGs - PEG 1500 and PEG 6000, both lineal but with different chain lengths, and PEG 12500 which was branched. The PEGs were deposited on silica plates using branched poly(ethylene imine) as an anchoring polymer. Fibroblasts were plated and studied by immunofluorescence to evaluate the overall cell morphology, the organisation of the actin cytoskeleton, and the beta1-integrin (fibronectin receptor). The particular effect of fibronectin (FN) pre-adsorption was studied. Our results suggest that PEG 6000 surface is to be preferable with respect to the initial interaction with the cells. The overall cell morphology was almost normal on bare surfaces. FN pre-coating additionally improved cell adhesion and spreading as well as the organization of the actin cytoskeleton and focal adhesion formation; the PEG 12500 surface showed relatively poor initial properties. Almost no cell spreading was found on the bare surface, but FN pre-adsorption completely restored normal cell morphology. In contrast, PEG 1500 had to be considered is 'the worst' material, because of lower initial cell adhesion and spreading and FN pre-adsorption did not restore normal cell morphology. PMID- 10385222 TI - Effect of Saiboku-to, an Oriental Herbal Medicine, on gastric lesion induced by restraint water-immersion stress or by ethanol treatment. AB - The effect of saiboku-to on gastric lesions induced by restraint water-immersion stress and ethanol has been examined in rats. Thirty minutes after oral administration of saiboku-to, the rats were placed in restraint cages and immersed in water at 23 degrees C for 7 h, or orally administered 99.5% ethanol (1 mL) and placed in normal cages for 1 h. The stress for 7 h or the ethanol treatment for 1h induced erosion in the glandular area of the stomach. Histology showed that the surface epithelial cells were desquamated and part of the lamina propria mucosae was injured. The evaluation of lesion index, the cumulative length of the gastric lesion, on the gross appearance of the stomach, revealed that saiboku-to dose-dependently inhibited both the water-immersion stress induced gastric erosion and ethanol-induced gastric erosion. To determine whether the anti-erosion effect of saiboku-to was because of a mild irritant effect, saiboku-to or 20% ethanol, which is known as a typical mild irritant, were given orally. After 30 min a strong irritant, 99.5% ethanol, was given orally. Histological examination was performed 30 min after administration of saiboku-to or the mild irritant, and 1 h after administration of the strong irritant. The mild irritant induced a reduction in surface epithelial cells 30 min after administration. Furthermore, the mild irritant protected the stomach against mucosal erosion produced by the strong irritant. Saiboku-to protected the strong irritant-induced erosion without producing mild irritation as observed in stomach treated with 20% ethanol. Pretreatment with saiboku-to also inhibited the decrease in the levels of hexosamine, gastric mucus glycoprotein, induced by the strong irritant. In pylorus-ligated rats, saiboku-to dose-dependently inhibited gastric acid secretion, a gastric aggressive factor. These results suggest that the anti-erosion effect of saiboku-to which is not a mild irritant, involves both inhibition of aggressive factors, such as gastric acid secretion, and augmentation of defensive factors, such as gastric mucus cells. PMID- 10385224 TI - Synthesis and enzymatic degradation of optically active depsipeptide copolymers. AB - This paper describes the synthesis and biodegradation of copolymers of cyclic depsipeptide with epsilon-caprolactone (CL) or lactide (LA). Optically active cyclic depsipeptides, 3,6-dimethyl-2,5-morpholinediones (DMOs), were prepared by the reaction of an amino acid (D-, L-, or DL-alanine) with a hydroxy acid derivative (DL-2-bromopropionyl bromide). These isomers are abbreviated as D-DMO, L-DMO and DL-DMO respectively, according to the names of alanine isomers. Then, we have prepared the copolymers of DMO isomers with CL using tin(II) octylate as a catalyst. The NMR spectra and thermal properties of DMO/CL copolymers revealed that these copolymers exist randomly. The enzymatic degradation of the copolymers has been examined using Rhizopus delemar lipase, cholesterol esterase (from Pseudomonas sp.), and Proteinase K (from Tritirachium album). Cholesterol esterase and Proteinase K show high degradability, while the lipase shows little degradation. Among the enzymes used, only Proteinase K could recognize the isomerism of DMO, resulting in the following order of degradability: copoly(L DMO/CL) > copoly(DL-DMO/CL) > copoly(D-DMO/CL), i.e. this enzyme has the highest substrate specificity for naturally occurring L-alanine. Further, we have prepared the random copolymers of L-DMO with lactide (L-LA or DL-LA), and evaluated the enzymatic degradation of the copolymers by Proteinase K. The introduction of a small amount (up to c. 10 mol%) of L-DMO unit into LA homopolymers brought about greater degradability compared with LA homopolymers. In particular, L-DMO/L-LA copolymers with high degradability have been obtained without significant decrease in the mechanical and thermal properties of L-LA homopolymer. PMID- 10385225 TI - The effect of surface roughness of microporous membranes on the kinetics of oxygen consumption and ammonia elimination by adherent hepatocytes. AB - In membrane hybrid liver support devices (HLSDs) using isolated hepatocytes where oxygen is transported only by diffusion to the cells, about 15-40% of the cell mass is likely to be in direct contact with the semipermeable membranes used as immunoselective barriers: quantitative effects of membrane surface properties on the kinetics of hepatocyte metabolic reactions may also affect HLSD performance. In this paper, we report our investigation of the effects of surface morphology of two microporous commercial membranes on the kinetics of oxygen consumption and ammonia elimination by primary hepatocytes in adhesion culture. Isolated rat hepatocytes were cultured on polypropylene microporous membranes with different surface roughness and pore size in a continuous-flow bioreactor whose fluid dynamics was optimized for the kinetic characterization of liver cell metabolic reactions. Collagen-coated membranes were used as the reference substratum. Hepatocyte adhesion was not significantly affected by membrane surface morphology. The rates of the investigated reactions increased with ammonia concentration according to saturation kinetics: the values of kinetic parameters Vmax and K(M) increased as cells were cultured on the membrane with the greatest membrane surface roughness and pore size. For the reaction of oxygen consumption, Vmax increased from 0.066 to 0.1 pmol h(-1) per cell as surface roughness increased from 70 to 370 nm. For the kinetics of ammonia elimination. K(M) increased from 0.23 to 0.32 mM and Vmax increased from 1.49 to 1.79 pmol h(-1) per cell with membrane surface roughness increasing from 70 to 370 nm. Cells cultured on collagen-coated membranes consistently yielded the highest reaction rates. The Vmax values of 0.18 and 2.84 pmol h(-1) per cell for oxygen consumption and ammonia elimination, respectively, suggest that cell functions are also affected by the chemical nature of the substratum. PMID- 10385226 TI - Fibronectin adsorption or/and covalent grafting on chemically modified PEEK film surfaces. AB - Poly(ether ether ketone) (PEEK) films were chemically modified, by surface wet chemistry, into PEEK-OH, PEEK-NH2, and PEEK-NCO. Fibronectin (FN) adsorption, in the presence or absence of two non-ionic surfactants, was compared onto PEEK, PEEK-OH, and PEEK-NH2 on which the protein can only be adsorbed, and onto PEEK NCO on which FN could be covalently grafted. The amounts of FN present on the various supports were assayed by ELISA and LSC (with 125I-labeled FN). The remarkable effect of Pluronic F68 in preventing non-specific protein adhesion on the less hydrophilic surfaces was pointed out. Accordingly, a procedure could be proposed that allows minimal FN adhesion vs FN fixation on PEEK-NCO. The resulting PEEK-FN film, which immobilized 120-150 ng FN cm(-2), constitutes a new substratum for cell cultivation. PMID- 10385227 TI - Bacterial adhesion to functionalized polyurethanes. AB - The effect of fibrinogen and high molecular weight kininogen on bacterial adhesion to functionalized polyurethanes was studied. Glass slides were coated with different polyurethanes, including Pellethane, sulfonated Pellethane, phosphonated Pellethane, a zwitterionic phosphonated polyurethane, and quaternized amine polyurethanes. The polymer-coated glass squares were exposed to radiolabelled S. aureus. When comparing adhesion to bare polyurethanes, it was found that adhesion was lowest on the phosphonated Pellethane and the zwitterionic phosphonated polyurethane while highest on the methyl quaternized polyurethanes. Fibrinogen-mediated adhesion was studied by first exposing the polymers to increasing concentrations of canine fibrinogen before incubating them with S. aureus. All the polymers except the quaternized amine polyurethanes exhibited at least ten-fold increases in bacterial adhesion as the fibrinogen treatment concentration was increased from 0.0 to 10.0 microg ml(-1). The quaternized amine polyurethanes maintained their relatively high amount of bacterial adhesion regardless of the fibrinogen concentration. The effect of two chain high molecular weight kininogen (TCHMWK) on fibrinogen-mediated bacterial adhesion was assessed by exposing the polymers to 1.0 microg ml(-1) fibrinogen followed by two different concentrations of TCHMWK. Decreases in bacterial adhesion were observed on all the polymers except the quaternized amine polyurethanes, which again retained their relatively high amount of bacterial adhesion. PMID- 10385228 TI - Suppressive effect of distinct bradykinin B2 receptor antagonist on allergen evoked exudation and leukocyte infiltration in sensitized rats. AB - 1. Bradykinin is suggested to play a role in the pathophysiology of several acute and chronic diseases, including allergic disorders such as asthma. In the present study, we have investigated the importance of bradykinin in mediating allergic inflammation in rats. 2. To this end we have tested the effects of the B2 receptor antagonists Hoe 140, FR173657 or FR172357 on the pleural inflammatory response triggered by intrapleural (i.pl.) injection of allergen (ovalbumin, 12 microg cavity(-1)) in 14 day-actively sensitized Wistar rats. Analysis of the pleural fluid effluent revealed a sequence of mast cell-dependent inflammatory events, including early protein exudation and neutrophilia and late pleural eosinophil influx. 3. Local treatment with Hoe 140 (0.1 and 1 microg cavity(-1)), FR173657 (1 and 10 microg cavity(-1)) or FR172357 (1 and 10 microg cavity(-1)) inhibited dose-dependently allergen-induced mast cell activation with impairment of pleural plasma leakage, neutrophil accumulation and late eosinophil influx. 4. Moreover, the B2 receptor antagonists also dose-dependently inhibited the allergic like inflammatory pleurisy triggered by bradykinin (50 microg cavity( 1)), which is characterized by acute mast cell degranulation, protein leakage and pleural eosinophil infiltration. 5. Taken together, our findings provide substantial evidence to suggest that bradykinin acting on its B2 receptors play a critical role in mediating allergic mast cell-dependent inflammation in rats, and suggest that B2 receptor antagonists may be useful therapeutically to control allergic dysfunction. PMID- 10385229 TI - Effects of mitoxantrone on action potential and membrane currents in isolated cardiac myocytes. AB - 1. The effects of mitoxantrone (MTO), an anticancer drug, on the membrane electrical properties of cardiac myocytes were investigated using the whole-cell clamp technique. 2. In isolated guinea-pig ventricular myocytes, 30 microM MTO induced a time-dependent prolongation of action potential duration (APD) which was occasionally accompanied by early afterdepolarizations. APD prolongation was preserved in the presence of 10 microM tetrodotoxin and showed reverse rate dependence. 3. Both the inward rectifier K+ current (I(KI)) and the delayed rectifier K+ current (I(K)) of guinea-pig ventricular myocytes were significantly depressed by 30 microM MTO. The rapidly activating component of I(k) (I(Kr)) seemed to be preferentially blocked by MTO. The transient outward current was not affected by MTO in rat ventricular myocytes. 4. Thirty microM MTO had no direct effect on the L-type Ca2+ current (I(Ca(L))), but reversed the inhibitory effect of 1 microM carbamylcholine but not the A1-adenosine receptor agonist (-)-N6 phenylisopropyladenosine (1 microM) on I(Ca(L)) enhanced by 50 nM isoprenaline in guinea-pig ventricular myocytes. In guinea-pig atrial mycotyes, 30 microM MTO inhibited by 93% the muscarinic receptor gated K+ current (I(K,ACh)) evoked by 1 microM carbamylcholine, whereas I(K,ACh) elicited by 100 microM GTPgammaS, a nonhydrolysable GTP analogue, was only decreased by 12%. 5. The specific binding of [3H]QNB, a muscarinic receptor ligand, to human atrial membranes was concentration-dependently displaced by MTO (1-1000 microM). 6. In conclusion, MTO blocks cardiac muscarinic receptors and prolongs APD by inhibition of I(KI) and I(Kr). The occasionally observed early afterdepolarizations may signify a potential cardiac hazard of the drug. PMID- 10385230 TI - Nitric oxide synthase inhibition by dimaprit and dimaprit analogues. AB - 1. The similarity in molecular structure between the histamine H2-agonist dimaprit (3-dimethylamino-propyl-isothiourea) and the endogenous nitric oxide synthase (NOS) substrate L-arginine prompted us to study the effect of dimaprit and some dimaprit analogues on NOS activity. Dimaprit and some of its analogues were tested in an in vitro assay which measures the conversion of [3H]-L-arginine to [3H]-L-citrulline. Dimaprit inhibits rat brain NOS (nNOS) concentration dependently with an IC50 of 49+/-14 microM. 2. Removal of one or both of the methyl groups from the non-isothiourea nitrogen of dimaprit improved nNOS inhibitory properties. Aminopropylisothiourea is the most potent compound (IC50 = 4.1+/-0.9 microM) of the series followed by methylaminopropylisothiourea (IC50 = 7.6 +/- microM). 3. The observed effect of aminopropylisothiourea and methylaminopropyl-isothiourea are probably not due to the compounds themselves but to the corresponding mercaptoalkylguanidines, rearrangement products formed in aqueous solutions. This hypothesis is strengthened by the finding that aminobutylisothiourea is not active since a rearrangement to mercaptobutylguanidine does not occur. 4. Remarkably, nitrosylation of the isothiourea group of dimaprit decreases nNOS inhibitory activity, while nitrosylation of the guanidine analogue of dimaprit increases the inhibition of nNOS activity. 5. The pharmacological profile of dimaprit includes inhibition of nNOS. The nNOS inhibitory activity occurs in the same concentration range as the H2-agonist and H3-agonist activity of this compound. PMID- 10385231 TI - The attenuation of learning impairments induced after exposure to CO or trimethyltin in mice by sigma (sigma) receptor ligands involves both sigma1 and sigma2 sites. AB - 1. Sigma (sigma) receptor ligands were previously reported to alleviate learning and memory impairments on several pharmacological and pathological rodent models of amnesia. Such effect was demonstrated as involving the sigma1 subtype of sigma receptor. 2. In this study, we characterized the pharmacological effect mediated by sigma ligands on two lesional models of amnesia in mice: (1) the hypoxia related learning and memory impairment model induced by repeated exposure to carbon monoxide (CO) gas; and (2) the intoxication with trimethyltin (1 mg kg( 1)). 3. The selective sigma1 ligand PRE-084 (1 mg kg(-1)) or the non-selective sigma1/sigma2 compounds DTG (0.1 mg kg(-1)), BD1008 (3 mg kg(-1)), and haloperidol (0.1 mg kg(-1)) reversed significantly the spontaneous alternation deficits observed 7 days after exposure to CO or 14 days after intoxication with trimethyltin. 4. The selective sigma1 receptor antagonist NE-100 (1 mg kg(-1)) was ineffective by itself, but blocked completely the PRE-084 effects, partially the DTG effects, and did not affect the effects induced by BD1008 or haloperidol. 5. A similar pharmacological profile was observed in the step-down type passive avoidance test performed 8 days after exposure to CO. 6. These results show that, in contrast to the previously reported amnesia models, the impairments induced after exposure to CO or intoxication with trimethyltin could be alleviated not only by sigma1 receptor agonists but also by sigma2 agonists. The particular pattern of neurodegeneration observed in these lesional models may explain these differences. PMID- 10385232 TI - Parallel modulation of receptor for activated C kinase 1 and protein kinase C alpha and beta isoforms in brains of morphine-treated rats. AB - 1. Receptor for activated C kinase 1 (RACK1) is an intracellular receptor for protein kinase C (PKC) that regulates the cellular enzyme localization. Because opiate drugs modulate the levels of brain PKC (Ventayol et al., 1997), the aim of this study was to assess in parallel the effects of morphine on RACK1 and PKC alpha and beta isozymes densities in rat brain frontal cortex by immunoblot assays. 2. Acute morphine (30 mg kg(-1), i.p., 2 h) induced significant increases in the densities of RACK1 (33%), PKC-alpha (35%) and PKC-beta (23%). In contrast, chronic morphine (10-100 mg kg(-1), i.p., 5 days) induced a decrease in RACK1 levels (22%), paralleled by decreases in the levels of PKC-alpha (16%) and PKC beta (16%). 3. Spontaneous (48 h) and naloxone (2 mg kg(-1), i.p., 2 h) precipitated morphine withdrawal after chronic morphine induced marked up regulations in the levels of RACK1 (38-41%), PKC-alpha (51-52%) and PKC-beta (48 62%). 4. In the same brains and for all combined treatments, there were significant positive correlations between the density of RACK1 and those of PKC alpha (r=0.85, n = 35) and PKC-beta (r=0.75, n=32). 5. These data indicate that RACK1 is involved in the short- and long-term effects of morphine and in opiate withdrawal, and that RACK1 modulation by morphine or its withdrawal is parallel to those of PKC-alpha and beta isozymes. Since RACK1 facilitates the PKC substrate accessibility, driving its cellular localization, the coordinate regulation of the PKC/RACK system by morphine could be a relevant molecular mechanism in opiate addiction. PMID- 10385233 TI - NO synthase in cholinergic nerves and NO-induced relaxation in the rat isolated corpus cavernosum. AB - 1. In the rat corpus cavernosum (CC), the distribution of immunoreactivity for neuronal and endothelial NO synthase (nNOS and eNOS), and the pattern of NOS immunoreactive (-IR) nerves in relation to some other nerve populations, were investigated. Cholinergic nerves were specifically immunolabelled with antibodies to the vesicular acetylcholine transporter protein (VAChT). 2. In the smooth muscle septa surrounding the cavernous spaces, and around the central and helicine arteries, the numbers of PGP- and tyrosine hydroxylase (TH)-IR terminals were large, whereas neuropeptide Y (NPY)-, VAChT-, nNOS-, and vasoactive intestinal polypeptide (VIP)-IR terminals were found in few to moderate numbers. 3. Double immunolabelling revealed that VAChT- and nNOS-IR terminals, VAChT- and VIP-IR terminals, nNOS-IR and VIP-IR terminals, and TH- and NPY-IR terminals showed coinciding profiles, and co-existence was verified by confocal laser scanning microscopy. TH immunoreactivity was not found in VAChT-, nNOS-, or VIP IR nerve fibres or terminals. 4. An isolated strip preparation of the rat CC was developed, and characterized. In this preparation, cumulative addition of NO to noradrenaline (NA)-contracted strips, produced concentration-dependent, rapid, and almost complete relaxations. Electrical field stimulation of endothelin-1 contracted preparations produced frequency-dependent responses: a contractile twitch followed by a fast relaxant response. After cessation of stimulation, there was a slow relaxant phase. Inhibition of NO synthesis, or blockade of guanylate cyclase, abolished the first relaxant phase, whereas the second relaxation was unaffected. 5. The results suggest that in the rat CC, nNOS, VAChT and VIP-immunoreactivities can be found in the same parasympathetic cholinergic neurons. Inhibitory neurotransmission involves activation of the NO-system, and the release of other, as yet unknown, transmitters. PMID- 10385235 TI - Structural and ionic determinants of 5-nitro-2-(3-phenylprophyl-amino)-benzoic acid block of the CFTR chloride channel. AB - 1. The goals of this study were to identify the structural components required for arylaminobenzoate block of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel and to determine the involvement of two positively charged amino acid residues, found within the channel, in drug binding. 2. Wild-type and mutant CFTR chloride channels were expressed in Xenopus oocytes and CFTR currents measured using the two microelectrode voltage clamp. Block of the wild-type CFTR current by 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB) occurred in a voltage-dependent manner with preferential inhibition of the inward currents (Kd = 166 microM at -90 mV). 3. Removal of the phenyl ring from the aliphatic chain of NPPB, with the compound 2-butylamino-5-nitrobenzoic acid, caused only a small change in CFTR inhibition (Kd = 243 microM), while addition of an extra phenyl ring at this position (5-nitro-2-(3,3-diphenylpropylamino) benzoic acid) increased drug potency (Kd = 58 microM). In contrast, removal of the benzoate ring (2-amino-4-phenylbutyric acid) or the 5-nitro group (2-(3 phenylpropylamino)-benzoic acid) of NPPB severely limited drug block of the wild type channel. 4. NPPB inhibition of CFTR currents in oocytes expressing the mutants K335E and R347E also occurred in a voltage-dependent manner. However, the Kds for NPPB block were increased to 371 and 1573 microM, for the K335E and R347E mutants, respectively. 5. NPPB block of the inward wild-type CFTR current was reduced in the presence of 10 mM of the permeant anion SCN-. 6. These studies present the first step in the development of high affinity probes to the CFTR channel. PMID- 10385234 TI - Characterization of [3H]-heparin binding in human vascular smooth muscle cells and its relationship to the inhibition of DNA synthesis. AB - 1. The glycosaminoglycan heparin inhibits vascular smooth muscle cell (VSMC) proliferation and migration, but the mechanism of its antiproliferative action remains unclear. Heparin has been reported to bind to high affinity cell surface sites on animal VSMC before undergoing receptor mediated endocytosis resulting in signal transduction into the cytoplasm and modulation of genes involved in proliferation. In this study, we have characterized the binding of [3H]-heparin to human saphenous vein-derived VSMC and examined whether there is any relationship between the affinity of [3H]-heparin binding and the inhibitory effect of heparin and its structural analogues on DNA synthesis. 2. At 4 degrees C [3H]-heparin binding to human VSMC occurred in a specific, time and concentration-dependent manner and was not influenced by the removal of calcium ions. Binding of the ligand appeared to occur to the cell surface and was both saturable and reversible. Kinetic and steady state data indicated a single class of binding sites. 3. The pharmacology of [3H]-heparin binding was examined in displacement studies using unlabelled heparin and structural analogues. A comparison of the rank potencies of heparin, heparan sulphate fraction II, low molecular weight heparin and trehalose octasulphate showed that there was a marked discrepancy between their estimated affinities in the binding assays and their effect on DNA synthesis. 4. In summary, we have characterized the heparin binding site on human saphenous vein-derived VSMC. Our findings suggest that the action of heparin and its analogues on DNA synthesis does not simply reflect an interaction with the cell-associated heparin binding site defined in these studies, but may also be determined by the internalization and metabolism of the glycosaminoglycan(s). PMID- 10385236 TI - Subunit mutations affect ethanol actions on GABA(A) receptors expressed in Xenopus oocytes. AB - 1. Mutations of specific amino acids were introduced in transmembrane domains (TM) of GABA(A) receptor alpha2, beta1 and gamma2L subunits. The effects of these mutations on the action of ethanol were studied using the Xenopus oocyte expression system and two-electrode voltage-clamp recording techniques. 2. Mutant alpha2 subunits containing S270I (TM2) or A291W (TM3) made the receptor more sensitive to GABA, as compared to wild-type alpha2beta1gamma2L receptor. The mutation S265I (TM2) of beta1 and S280I (TM2) or S30IW (TM3) in gamma2L subunits did not alter apparent affinity of the receptor for GABA. M286W (TM3) in the beta1 subunit resulted in a receptor that was tonically open. 3. Using an EC5 concentration of GABA, the function of the wild-type receptor with alpha2beta1gamma2L subunits was potentiated by ethanol (50-200 mM). The mutations in TM2 or TM3 of the alpha2 subunit diminished the potentiation by ethanol. The action of ethanol was also eliminated with a mutation in the TM2 site of the beta1 subunit. Ethanol produced significant inhibition of GABA responses in receptors containing the combination of alpha2 and beta1 TM2 mutants with a wild type gamma2L subunit. A small but significant reduction in the potentiation by ethanol was observed with gamma2L TM2 and/or TM3 mutants. 4. From these results, we suggest that in heteromeric GABA(A) receptors composed of the alpha, beta and gamma subunits, ethanol may bind in a cavity formed by TM2 and TM3, and that binding to the alpha or beta subunit may be more critical than the gamma subunit. PMID- 10385237 TI - Neuropeptide Y is a prejunctional inhibitor of vagal but not sympathetic inotropic responses in guinea-pig isolated left atria. AB - 1. The effects of NPY and related peptides were examined on basal contractile force and nerve-mediated inotropic responses to electrical field stimulation of the guinea-pig isolated left atrium. 2. Electrical field stimulus (EFS)-inotropic response curves were constructed by applying 1-64 trains of four field pulses (200 Hz, 0.1 ms duration, 100 V) across isolated left atria (paced at 4 Hz, 2 ms, 1-4 V) within the atrial refractory period. Curves were constructed in presence of vehicle, propranolol (1 microM) or atropine (1 microM) to determine appropriate stimulus conditions. 3. The effects of PYY (1-10,000 nM), NPY (0.01 10 microM), N-Ac-[Leu28,31]NPY(24-36) (N-A[L]NPY(24-36); 0.01-10 microM) and clonidine (0.1-1000 nM) were examined on the positive and negative inotropic responses to EFS (eight trains, four pulses per refractory period). 4. NPY related peptides had no effect on basal force of contraction nor on the inotropic concentration-response curves to bethanechol or isoprenaline. All three peptides inhibited vagally-mediated negative inotropic responses; rank order of potency PYY>NPY> or =N-A[L]NPY(24-36) was consistent with an action at prejunctional Y2 receptors. Clonidine concentration-dependently inhibited sympathetic inotropic responses. However, PYY, NPY and N-A[L]NPY(24-36) failed to mediate any significant inhibition of the positive inotropic response to EFS. 5. These data demonstrate that NPY is an effective inhibitor of vagal but not sympathetically mediated inotropic responses in the guinea-pig isolated left atria. This may suggest that endogenously co-released NPY is important in mediating cross talk between efferent components of the autonomic nervous system modulating cardiac contractility, acting overall to sustain positive inotropic responses. PMID- 10385238 TI - Hyperosmolarity reduces the relaxing potency of nitric oxide donors in guinea-pig trachea. AB - 1. Non-responders to inhaled nitric oxide treatment have been observed in various patient groups. The bronchodilatory effect of inhaled nitric oxide was attenuated when the airway lumen was rendered hyperosmolar in an in vivo study on rabbits. We used a guinea-pig tracheal perfusion model to investigate the effects of increased osmolarity (450 mOsm, NaCl added) on the relaxing potency of the nitric oxide donors sodium nitroprusside (SNP) and (+/-)-S-nitroso-N-acetylpenicillamine (SNAP). 2. Under iso-osmolar conditions SNP relaxed the carbachol (CCh, 1 microM) contracted trachea by 83+/-3%. After pretreatment with intraluminal hyperosmolarity SNP relaxed the CCh-contracted trachea by only 31+/-7% (P<0.05). When the trachea was contracted to the same extent under untreated and hyperosmolar conditions, the untreated trachea was completely relaxed by SNP but, after hyperosmolar pretreatment, SNP could no longer relax the trachea. 3. SNAP relaxed the CCh contracted trachea by 27+/-5%. After pretreatment with intraluminal hyperosmolarity, SNAP relaxed the trachea by 11+/-4%, which was less than in the iso-osmolar control (P<0.05). 4. Extraluminal hyperosmolarity did not affect carbachol elicited contraction, and SNP administered externally during extraluminal hyperosmolarity was able to relax the trachea (P<0.05). 5. The cell permeable guanosine 3'5'-cyclic monophosphate analogue 8-Br-cGMP relaxed the CCh contracted trachea in both iso-osmolar (P<0.05) and hyperosmolar conditions (P<0.05). 6. The relaxant effect of nitric oxide donors on tracheal smooth muscle is markedly reduced when the airway epithelium is exposed to hyperosmolar solution. PMID- 10385239 TI - Antibodies and antisense oligodeoxynucleotides to mu-opioid receptors, selectively block the effects of mu-opioid agonists on intestinal transit and permeability in mice. AB - 1. We have studied the effects of mu and delta opioids on intestinal function (permeability, PER; gastrointestinal transit, GIT), and their antagonism after the intracerebroventricular (i.c.v.) administration of specific antibodies (ABs) or antisense oligodeoxynucleotides (ODN) to mu-receptors (OR). Central versus peripheral site/s of action of subcutaneous (s.c.) mu-opioids, were also assessed. 2. Male Swiss CD-1 mice were used. GIT was measured with charcoal and PER by the passage of 51Cr-EDTA from blood to lumen. 3. Morphine and fentanyl (i.c.v. and s.c.) inhibited GIT and PER in a dose-related manner; they were more potent by i.c.v. route, both on GIT and PER (70 and 17 times for morphine and fentanyl). They also had a greater effect on GIT than PER (4.3 and 1.6 times). DPDPE had a lower potency than mu-agonists in all experiments, and no dose response could be obtained after s.c. administration on GIT. 4. Pretreatment with i.c.v. ABs (24 h) or antisense ODN (5 days), decreased the effects (GIT and PER) of i.c.v. morphine and fentanyl, while those of DPDPE remained unchanged. The ABs did not alter the peripheral effects of mu-opioids. 5. The results show that (i.c.v. or s.c.) mu opioids produce dose-related inhibitions of PER and GIT, being more potent by the i.c.v. route. Delta-opioids had a greater effect on PER than GIT, while the opposite occurred for mu-agonists. Pretreatment with ABs or ODN to mu-OR, blocked the central effects of mu (but not delta) agonists on GIT and PER. PMID- 10385240 TI - Investigation of the role of 5-HT1B and 5-HT1D receptors in the sumatriptan induced constriction of porcine carotid arteriovenous anastomoses. AB - 1. It has previously been shown that the antimigraine drug sumatriptan constricts porcine carotid arteriovenous anastomoses via 5-HT1-like receptors, identical to 5-H1B/1D receptors. The recent availability of silent antagonists selective for the 5-HT1B (SB224289) and 5-HT1D (BRL15572) receptor led us to further analyse the nature of receptors involved. 2. In pentobarbitone-anaesthetized, bilaterally vagosympathectomized pigs, sumatriptan (30, 100 and 300 microg kg(-1), i.v.) dose dependently decreased carotid arteriovenous anastomotic conductance by up to 70+/ 5%. 3. The dose-related decreases in carotid arteriovenous anastomotic conductance by sumatriptan (30, 100 and 300 microg kg(-1), i.v.) remained unchanged in animals treated (i.v.) with 1 mg kg(-1) of BRL15572 (maximum decrease: 72+/-3%), but were significantly attenuated by 1 mg kg(-1) (maximum decrease: 30+/-11%) and abolished by 3 mg kg(-1) (maximum decrease: 3+/-7%) of SB224289. The highest dose of SB224289 did not attenuate the hypertension, tachycardia or increases in carotid blood flow induced by bolus injections of noradrenaline (0.1-3 microg kg(-1), i.v.). 4. The results indicate that sumatriptan constricts porcine carotid arteriovenous anastomoses primarily via 5 HT1B, but not via 5-HT1D receptors. PMID- 10385241 TI - Pharmacological characterization of the muscarinic receptor antagonist, glycopyrrolate, in human and guinea-pig airways. AB - 1. In this study we have evaluated the pharmacological profile of the muscarinic antagonist glycopyrrolate in guinea-pig and human airways in comparison with the commonly used antagonist ipratropium bromide. 2. Glycopyrrolate and ipratropium bromide inhibited EFS-induced contraction of guinea-pig trachea and human airways in a concentration-dependent manner. Glycopyrrolate was more potent than ipratropium bromide. 3. The onset of action (time to attainment of 50% of maximum response) of glycopyrrolate was similar to that obtained with ipratropium bromide in both preparations. In guinea-pig trachea, the offset of action (time taken for response to return to 50% recovery after wash out of the test antagonist) for glycopyrrolate (t1/2 [offset]=26.4+/-0.5 min) was less than that obtained with ipratropium bromide (81.2+/-3.7 min). In human airways, however, the duration of action of glycopyrrolate (t1/2 [offset]>96 min) was significantly more prolonged compared to ipratropium bromide (t1/2 [offset]= 59.2+/-17.8 min). 4. In competition studies, glycopyrrolate and ipratropium bromide bind human peripheral lung and human airway smooth muscle (HASM) muscarinic receptors with affinities in the nanomolar range (K1 values 0.5-3.6 nM). Similar to ipratropium bromide, glycopyrrolate showed no selectivity in its binding to the M1-M3 receptors. Kinetics studies, however, showed that glycopyrrolate dissociates slowly from HASM muscarinic receptors (60% protection against [3H]-NMS binding at 30 nM) compared to ipratropium bromide. 5. These results suggest that glycopyrrolate bind human and guinea-pig airway muscarinic receptors with high affinity. Furthermore, we suggest that the slow dissociation profile of glycopyrrolate might be the underlying mechanism by which this drug accomplishes its long duration of action. PMID- 10385242 TI - Role of potassium channels and nitric oxide in the relaxant effects elicited by beta-adrenoceptor agonists on hypoxic vasoconstriction in the isolated perfused lung of the rat. AB - 1. The aims of this study were to compare, in the rat isolated perfused lung preparation, the antagonist effects of a nonselective beta-adrenoceptor agonist (isoprenaline), a selective beta2-adrenoceptor agonist (salbutamol) and a selective beta3-adrenoceptor agonist (SR 59104A) on the hypoxic pulmonary pressure response, and to investigate the role of K+ channels, endothelium derived relaxing factor and prostaglandins in these effects. K+ channels were inhibited by glibenclamide, charybdotoxin or apamin, NO synthase and cyclo oxygenase were inhibited by N(G)-nitro-L-arginine methyl ester (L-NAME) and indomethacin, respectively. 2. Hypoxic ventilation produced a significant increase in perfusion pressure (+65%, P<0.001) and L-NAME significantly increased this response further (+123%, P<0.01). After apamin, L-NAME, indomethacin, post hypoxic basal pressure did not return to baseline values (P<0.001). 3. Glibenclamide partially inhibited the relaxant effects of isoprenaline (P<0.05) and salbutamol (P<0.001) but not that of SR 59104A. In contrast, charybdotoxin and apamin partially inhibited the relaxant effects of SR 59104A (P=0.053 and <0.01, respectively) but did not modify the effects of isoprenaline and salbutamol. L-NAME partially inhibited the dilator response of salbutamol (P<0.01) and SR 59104A (P<0.05) but not that of isoprenaline. 4. We conclude that (a) EDRF exerts a significant inhibition of the hypoxic pulmonary response, (b) SK(Ca) channel activation, EDRF and prostaglandins contribute to the reversal of the hypoxic pressure response, (c) the vasodilation induced by isoprenaline is mediated in part by activation of K(ATP) channels, that of salbutamol by activation of K(ATP) channels and EDRF. In contrast, SR 59104A partly operates through BK(Ca), SK(Ca), channels and EDRF activation, differing in this from the beta1 and beta2-adrenoceptor agonists. PMID- 10385243 TI - Antiarrhythmic effect and its underlying ionic mechanism of 17beta-estradiol in cardiac myocytes. AB - 1. The effects of oestrogens on action potential and membrane currents were examined in single guinea-pig atrial myocytes. 2. 17Beta-estradiol (3-10 microM) shortened the action potential duration without significant changes in the resting membrane potential. E-4031 (1 microM) markedly prolonged the action potential duration and induced early afterdepolarization, and 17beta-estradiol (10 microM) abolished it. 3. When cells were perfused in isoproterenol-containing solution, action potentials due to abnormal automaticity caused by membrane depolarization developed, and were also inhibited by 17beta-estradiol. 4. Under voltage clamp conditions, the voltage-dependent Ca2+ currents consisted of both T (I(Ca,T)) and L-type (I(Ca,L)). 17Beta-estradiol reduced I(Ca,L) concentration dependently, while it (10 microM) suppressed I(Ca,T) only by approximately 10%. 17Beta-estradiol did not affect time courses of I(Ca,L) inactivation, but it shifted the steady-state inactivation curve to more negative potentials. 5. 17Beta-estradiol (10 microM) did not affect the time-dependent K+ current (I(K)), referred to as I(Kr) and I(Ks) and inwardly rectifying K+ current. However, 17beta-estradiol (30 microM) or diethylstilbestrol (10 microM) inhibited K+ currents. 6. DES and ethinylestradiol (EES) also suppressed I(Ca,L), but testosterone and progesterone failed to inhibit I(Ca,L) The potency of the inhibitory effect on I(Ca,L) was DES> EES> 17beta-estradiol. 7. 17Beta-estradiol and DES also inhibited the cyclic AMP-enhanced I(Ca,L), but cyclic GMP in the pipette or pretreatment of L-NAME could not block the effects of oestrogen on I(Ca,L). 8 These results suggest that oestrogen specifically has antiarrhythmic effects, possibly by acting the L-type Ca2+ channels. The antiarrhythmic effects of oestrogens may contribute to the cardioprotective actions of oestrogens. PMID- 10385244 TI - Interleukin-6 production by activated human monocytic cells is enhanced by MK 571, a specific inhibitor of the multi-drug resistance protein-1. AB - 1. The intracellular transport of leukotriene C4 (LTC4) in hematopoietic cells such as human monocytes is controlled by an ATP dependent carrier encoded by the multidrug resistance protein1 (MRPI) gene whose function can be blocked by the compound MK-571. Since LTs play a major role in control of cytokine expression in monocytes, we questioned whether blocking of the MRPI mediated function by MK-571 might affect cytokine production. 2. MK-571 strongly enhanced IL-6 expression at mRNA and protein level in lipopolysaccharide (LPS) and interleukin-1 (IL-1) stimulated human monocytes giving rise to 2.0+/-0.4 (x+/-s.d.) and 5.7+/-3.5 fold induction of IL-6 protein secretion. The increase in IL-6 secretion was accompanied by an enhanced phosphorylation of p38 but not of c-Jun-N terminal kinase. 3. The involvement of the kinase signalling pathways was further analysed by using SB203580 and PD98059, specific inhibitors of the p38 and ERK1/2 signalling route. MK-571 mediated upregulation of IL-6 in the presence of IL-1 was partially attenuated by SB203580 and PD98059. Electrophoretic mobility shift assays demonstrated that MK-571 did not affect the IL-1 induced DNA binding activity of Activator Protein-1 and Nuclear Factor-kappaB but rather enhanced the transactivational activity of an IL-6 promoter construct. Finally it was shown that the MK-571 mediated effects on IL-6 secretion could not be inhibited by the LT synthesis inhibitor SB203347 or by the anti-oxidant pyrrolidine dithiocarbamate (PDTC). 4. These results indicate that the membrane transporter MRP1 is involved in the regulation of IL-6 expression in activated human peripheral blood monocytes. PMID- 10385245 TI - In vivo pathway of thermal hyperalgesia by intrathecal administration of alpha,beta-methylene ATP in mouse spinal cord: involvement of the glutamate-NMDA receptor system. AB - 1. The aim of the present study is to characterize the role of the P2X receptor in spinal nociceptive processing in vivo. We investigated the mechanisms of the P2X receptor agonist alpha,beta-methylene ATP (alpha,betameATP)-induced modulation of acute nociceptive signalling in mouse spinal cord. 2. Intrathecal administration of alpha,betameATP produced a significant and dose-dependent thermal hyperalgesic response. This response was completely blocked by intrathecal pretreatment with the non-selective P2 receptor antagonist, pyridoxal phosphate-6-azophenyl-2',4'-disulphonate (PPADS) and the selective P2X1, P2X3 and P2X2-3 receptor antagonist, 2',3'-O-(2,4,6-trinitrophenyl)adenosine 5' triphosphate (TNP-ATP). Pretreatment with alpha,betameATP 15, 30 and 60 min prior to administration of a second dose of alpha,betameATP diminished the alpha,betameATP-induced thermal hyperalgesia. 3. A potent agonist for the P2X1 receptor, beta,gamma-methylene-L-ATP, did not show the hyperalgesic response, indicating that the P2X1 receptor is not involved in the spinal nociceptive pathway. 4. In fura-2 experiments using mouse dorsal root ganglion (DRG) neurons, alpha,betameATP (100 microM) increased intracellular Ca2+ ([Ca2+]i). This was not produced by a second application of alpha,betameATP. The same DRG neurons also showed a marked [Ca2+]i increase in response to capsaicin (3 microM). 5. Intrathecal pretreatment with the Ca2+-dependent exocytosis inhibitor, botulinum neurotoxin B, abolished the thermal hyperalgesia by alpha,betameATP. Furthermore, thermal hyperalgesia was significantly inhibited by the N-methyl-D-aspartate (NMDA) receptor antagonists, 2-amino-5-phosphonopentanoate (APV), dizocilpine and ifenprodil. 6. These findings suggest that alpha,betameATP-induced thermal hyperalgesia may be mediated by the spinal P2X3 receptor subtype that causes unresponsiveness by repetitive agonist applications, and that alpha,betameATP (perhaps through P2X3 receptors) may evoke spinal glutamate release which, in turn, leads to the generation of thermal hyperalgesia via activation of NMDA receptors. PMID- 10385246 TI - Capsaicin-insensitive sensory-efferent meningeal vasodilatation evoked by electrical stimulation of trigeminal nerve fibres in the rat. AB - 1. Antidromic vasodilatation and plasma extravasation to stimulation of the trigeminal ganglion or its perivascular meningeal fibres was investigated by laser-Doppler flowmetry and 125I-labelled bovin serum albumin in the dura mater and in exteroceptive areas (nasal mucosa, upper eyelid) of anaesthetized rats pretreated with guanethidine and pipecuronium. 2 Trigeminal stimulation at 5 Hz for 20 s elicited unilateral phasic vasodilatation in the dura and lasting response in the nasal mucosa. Resiniferatoxin (1-3 microg kg(-1) i.v.), topical (1%) or systemic capsaicin pretreatment (300 mg kg(-1) s.c. plus 1 mg kg(-1) i.v.) did not inhibit the meningeal responses but abolished or strongly inhibited the nasal responses. Administration of vinpocetine (3 mg kg(-1) i.v.) increased both basal blood flow and the dural vasodilatation to perivascular nerve stimulation. 3. Dural vasodilatation to trigeminal stimulation was not inhibited by the calcitonin gene-related peptide-1 receptor (CGRP-1) antagonist hCGRP8-37 (15 or 50 microg kg(-1) i.v), or the neurokinin-1 receptor antagonist RP 67580 (0.1 mg kg(-1) i.v.) although both antagonists inhibited the nasal response. Neither mucosal nor meningeal responses were inhibited by atropine (5 mg kg(-1) i.v.), hexamethonium (10 mg kg(-1) i.v.) or the vasoactive intestinal polypeptide (VIP) antagonist (p-chloro-D-Phe6-Leul7)VIP (20 microg kg(-1) i.v.). 4. Plasma extravasation in the dura and upper eyelid elicited by electrical stimulation of the trigeminal ganglion was almost completely abolished in rats pretreated with resiniferatoxin (3 microg kg(-1) i.v.). 5. It is concluded that in the rat meningeal vasodilatation evoked by stimulation of trigeminal fibres is mediated by capsaicin-insensitive primary afferents, while plasma extravasation in the dura and upper eyelid and the vasodilatation in the nasal mucosa are mediated by capsaicin-sensitive trigeminal fibres. PMID- 10385247 TI - Differential pharmacology between the guinea-pig and the gorilla 5-HT1D receptor as probed with isochromans (5-HT1D-selective ligands). AB - 1. Both the 5-HT1D and 5-HT1B receptors are implicated in migraine pathophysiology. Recently isochromans have been discovered to bind primate 5-HT1D receptors with much higher affinity than 5-HT1B receptors. In the guinea-pig, a primary animal model for anti-migraine drug testing, however, isochromans bound the 5-HT1D receptor with lower affinity than the gorilla receptor. 2. This species-specific pharmacology was investigated, using site-directed mutagenesis on cloned guinea-pig receptors heterologously expressed in human embryonic kidney 293 cells. Mutations of threonine 100 and arginine 102 at the extracellular side of transmembrane II of the guinea-pig 5-HT1D receptor to the corresponding primate residues, isoleucine and histidine, respectively, enhanced its affinity for isochromans to that of the gorilla receptor, with little effects on its affinities for serotonin, sumatriptan and metergoline. Free energy change from the R102H mutation was about twice as much as that from the T100I mutation. 3. For G protein-coupling, serotonin marginally enhanced GTPgamma35S binding in membranes expressing the guinea-pig 5-HT1D receptor and its mutants, but robustly in membranes expressing the gorilla receptor. Sumatriptan enhanced GTPgamma35S binding in the latter nearly as much as serotonin, and several isochromans by 30 60% of serotonin. 4. We discovered key differences in the function and binding properties of guinea-pig and gorilla 5-HT1D receptors, and identified contributions of I100 and H102 of primate 5-HT1D receptors to isochroman binding. Among common experimental animals, only the rabbit shares I100 and H102 with primates, and could be useful for studying isochroman actions in vivo. PMID- 10385248 TI - Respiratory action of capsaicin microinjected into the nucleus of the solitary tract: involvement of vanilloid and tachykinin receptors. AB - 1. The respiratory response to microinjection of capsaicin into the commissural nucleus of the solitary tract (cNTS) of urethane-anaesthetized rats was investigated in the absence and presence of the competitive vanilloid (capsaicin) antagonist, capsazepine, and selective tachykinin NK1, NK2 and NK3 antagonists (RP 67580, SR 48968 and SR 142801, respectively). 2. Microinjection of capsaicin reduced respiratory frequency but not tidal volume (VT), leading to an overall reduction in minute ventilation (VE). The effect was dose-dependent between 0.5 and 2 nmol capsaicin. Doses greater than 2 nmol produced apnoea. Tachyphylaxis was observed following repeated injection of capsaicin (1 nmol, 30 min apart). 3. Capsazepine (1 nmol) had no effect on frequency or VT when injected alone but completely blocked the respiratory response to capsaicin (1 nmol). 4. RP 67580 (1 but not 5 nmol) alone depressed frequency and VT slightly. Moreover, RP 67580 appeared to potentiate the bradypnoeic effect of capsaicin. In contrast, SR 48968 and SR 142801 (1 and 5 nmol) alone had no significant effect on respiration. However, both agents significantly attenuated the reduction in frequency produced by capsaicin. 5. In conclusion, microinjection of capsaicin into the cNTS decreases overall ventilation, primarily by reducing frequency. The action of capsaicin appears from the data to be mediated by vanilloid receptors since it is blocked by the competitive vanilloid antagonist capsazepine and is subject to tachyphylaxis. However, since NK2 (SR 48968) and NK3 (SR 142801) receptor antagonists block the actions of capsaicin, we propose that capsaicin acts also by releasing tachykinins from central afferent terminals in the cNTS. PMID- 10385249 TI - Recombinant human erythropoietin inhibits iNOS activity and reverts vascular dysfunction in splanchnic artery occlusion shock. AB - 1. We investigated the effects of recombinant human erythropoietin (rh-EPO) in splanchnic artery occlusion (SAO) shock. Sham operated animals were used as controls. Survival rate, mean arterial blood pressure (MAP), serum Tumor Necrosis Factor (TNF-alpha), plasma nitrite/nitrate concentrations, red blood cell (RBC) count, blood haemoglobin (Hb), the responsiveness of aortic rings to phenylephrine (PE, 1 nM-10 microM) and the activity of inducible nitric oxide synthase (iNOS) were studied. 2. SAO shocked rats had a decreased survival rate (0% at 4 h of reperfusion, while sham shocked rats survived more than 4 h), enhanced serum TNF-alpha concentrations, increased plasma nitrite/nitrate levels (60+/-9.5 microM; sham shocked rats= 2+/-0.4 microM), decreased MAP, unchanged RBC count and blood Hb and enhanced iNOS activity in the aorta. Moreover aortic rings from shocked rats showed a marked hyporeactivity to PE. 3. Rh-EPO (25, 50 and 100 U 100 g(-1), 5 min following the onset of reperfusion) increased survival rate (70% at 4 h of reperfusion with the highest dose), reduced plasma nitrite/nitrate concentrations (10.3+/-3.3 microM), increased MAP, did not change RBC count and blood Hb, and inhibited iNOS activity in thoracic aortae. Furthermore rh-EPO, either in vivo or in vitro (10 U for 1 h in the organ bath), restored to control values the hyporeactivity to PE. Finally rh-EPO inhibited the activity of iNOS in peritoneal macrophages activated with endotoxin. 4. Our data suggest that rh-EPO protects against SAO shock by inhibiting iNOS activity. PMID- 10385251 TI - Tacrolimus suppresses tumour necrosis factor-alpha and protects against splanchnic artery occlusion shock. AB - 1. Tumour necrosis factor (TNF-alpha) is a pleiotropic cytokine which is deeply involved in the pathogenesis of splanchnic artery occlusion (SAO) shock. Tacrolimus, formerly known as FK506, is a macrolide antibiotic, that blocks the transcription of several proinflammatory cytokines including TNF-alpha. 2. Male anaesthetized rats were subjected to clamping of the splanchnic arteries for 45 min. This surgical procedure resulted in an irreversible state of shock (SAO shock). Sham operated animals were used as controls. SAO shocked rats had a decreased survival rate (0% at 4 h of reperfusion, while sham shocked rats survived more than 4 h), enhanced serum TNF-alpha concentrations (415+/-12 U ml( 1)), decreased mean arterial blood pressure (MAP), leukopenia and increased ileal leukocyte accumulation studied by means of myeloperoxidase activity (MPO=7.5+/ 0.3 U g(-1) tissue). Moreover aortic rings from shocked rats showed a marked hyporeactivity to phenylephrine (PE, 1 nM - 10 microM), reduced responsiveness to acetylcholine (ACh, 10 nM - 10 microM) and increased staining for intercellular adhesion molecule-1 (ICAM-1). Furthermore increased mRNA for TNF-alpha was observed in peritoneal macrophages of SAO shocked rats. 3. Tacrolimus (100 microg kg(-1), 5 min after splanchnic arteries occlusion) increased survival rate (SAO + Tacrolimus = 100% at 4 h of reperfusion), reverted the marked hypotension, reduced serum TNF-alpha (15+/-3 U ml(-1)), ameliorated leukopenia, reduced ileal MPO (0.9+/-0.01 U g(-1) tissue), restored to control values the hyporeactivity to PE. improved the reduced responsiveness to ACh and blunted the enhanced immunostaining for ICAM-1 in the aorta. Finally tacrolimus suppressed cytokine mRNA levels in peritoneal macrophages. 4. The data suggest that tacrolimus may represent a new therapeutic approach in circulatory shock. PMID- 10385250 TI - Synergistic effect with Phe-Gly-Leu-Met-NH2 of the C-terminal of substance P and insulin-like growth factor-1 on epithelial wound healing of rabbit cornea. AB - 1. We previously reported that substance P and insulin-like growth factor-1 (IGF 1) synergistically stimulate corneal epithelial wound healing in vitro and in vivo. We wished to identify which portion of the amino acid sequence of substance P might be responsible for this synergism. 2. Corneal epithelial migration was not affected by the addition of any one of the following factors: substance P; Phe-Gly-Leu-Met-NH2 (C-terminal of substance P); Val-Gly-Leu-Met-NH2 (C-terminal of neurokinin A, neurokinin B, and kassinin); Tyr-Gly-Leu-Met-NH2 (C-terminal of physalaemin); Ile-Gly-Leu-Met-NH2 (C-terminal of eledoisin); or Gly-Leu-Met-NH2 (common C-terminal of tachykinins). 3. In the presence of IGF-1, only substance P and Phe-Gly-Leu-Met-NH2 were synergistic in stimulating corneal epithelial migration in a dose-dependent fashion. 4. The combination of Phe-Gly-Leu-Met-NH2 and IGF-1 did not affect the incorporation of [3H]-thymidine into corneal epithelial cells. 5. Treatment with Phe-Gly-Leu-Met-NH2 and IGF-1, but not with Phe-Gly-Leu-Met-NH2 or IGF-1 alone, increased attachment of corneal epithelial cells to a fibronectin matrix. 6. The levels of alpha5 and beta1 integrin were not affected by Phe-Gly-Leu-Met-NH2 or IGF-1 alone, but they were significantly increased by the combination of Phe-Gly-Leu-Met-NH2 and IGF-1. 7. Topical application of the same combination facilitated corneal epithelial wound closure in vivo. 8. These results demonstrated that Phe-Gly-Leu-Met-NH2, a sequence of 4 amino-acids of the C-terminal of substance P, is the minimum sequence necessary to produce the synergistic effects of substance P and IGF-1 on corneal epithelial wound healing. PMID- 10385253 TI - Tonic inhibitory action by nitric oxide on spontaneous mechanical activity in rat proximal colon: involvement of cyclic GMP and apamin-sensitive K+ channels. AB - 1. The cellular mechanisms by which endogenous nitric oxide (NO) modulates spontaneous motility were investigated in rat isolated proximal colon. The mechanical activity was detected as changes in intraluminal pressure. 2. Apamin (1-100 nM) produced a concentration-dependent increase in the amplitude of the spontaneous pressure waves. The maximal contractile effect was of the same degree as that produced by Nomega-nitro-L-arginine methyl ester (L-NAME) (100 microM) and the joint application of apamin plus L-NAME had no additive effects. Apamin (0.1 microM) reduced the inhibitory effects (i.e. reduction in the amplitude of the pressure waves) induced by sodium nitroprusside (SNP) (1 nM - 10 microM) or 8 Br-cyclic GMP (1-100 microM). 3. 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) (0.1-5 microM), inhibitor of NO-stimulated guanylate cyclase, produced a concentration-dependent increase of the spontaneous contractions. ODQ (1 microM) in the presence of apamin (0.1 microM) did not produce any further increase in the contraction amplitude, whereas after L-NAME (100 microM) it decreased the spontaneous contractions. ODQ (1 microM) reduced the SNP inhibitory effects. 4. Zaprinast (1-50 microM), inhibitor of cyclic GMP phosphodiesterase, produced a concentration-dependent decrease of the spontaneous contractions. The effects of zaprinast were significantly reduced in the presence of apamin (0.1 microM) or L NAME (100 microM). 5. These results suggest that small conductance Ca2+-dependent K+ channels and cyclic GMP are involved in the modulation of the spontaneous contractile activity in rat proximal colon. Cyclic GMP production system and opening of apamin-sensitive K+ channels appear to work sequentially in transducing an endogenous NO signal. PMID- 10385252 TI - Endothelium is required in the vascular spasm induced by tetraethylammonium and endothelin-1 in guinea-pig aorta. AB - 1. To investigate the role of endothelium in vascular spasm, we studied the influence of endothelin-1 (ET-1) on the contracting and spasmogenic effect of the K+-channel blocker, tetraethylammonium (TEA), in aorta rings of reserpine-treated guinea-pigs, perfused with either control (5.5 mM) or elevated (50 mM) glucose concentration. 2. Endothelium-dependent relaxation induced by acetylcholine was lost in rings contracted by noradrenaline in the presence of elevated glucose. In control medium, TEA (1-20 mM) induced a sustained tonic contraction, followed by a phasic spasm, characterized by rhythmic contractions. Elevated glucose, ET-1 (3 nM), or both, reduced the EC50 of TEA-induced tonic contraction, without modifying the maximum contractile effect. 3. In control medium, ET-1 reduced the time before TEA-induced spasm and increased the rate of rhythmic contractions. TEA-induced spasm was abolished by elevated glucose, and restored by ET-1. The spasm induced by TEA and ET-1 was amplified by the ETA antagonist, EMD94246, and suppressed by the ET(A)-ET(B) antagonist, bosentan. In endothelium-denuded vessels incubated with high glucose and ET-1, TEA evoked only a tonic contraction. 4. In control medium, L-NAME (N(G)-nitro-L-arginine methyl ester) abolished TEA-induced rhythmic contractions. L-arginine, but not D-arginine, prevented the effect of L-NAME. In the presence of elevated glucose and ET-1, TEA induced spasm was not affected by L-NAME, whereas verapamil, indomethacin, metyrapone, glybenclamide or apamin abolished the phasic spasm, unmasking the tonic contracture. 5. In conclusion, endothelium plays a regulatory role in the genesis and maintenance of TEA-induced rhythmic contractions, through the release endothelium derived relaxing factor and vasodilating eicosanoids. PMID- 10385254 TI - Cyclic AMP suppresses interleukin-5 synthesis by human helper T cells via the downregulation of the calcium mobilization pathway. AB - 1. To delineate the mechanism by which cyclic AMP (cAMP) suppresses interleukin (IL)-5 synthesis, the effects of prostaglandin (PG) E2, forskolin, dibutyryl (db) cAMP and the Ca2+ ionophore, ionomycin on cytokine synthesis, proliferation and CD25 expression of human T cells were investigated. Further studies were performed by measurement of the intracellular concentrations of cyclic AMP ([cAMP]i) and Ca2+ ([Ca2+]i) and by electrophoretic mobility shift analysis (EMSA). 2. PGE2, forskolin and db-cAMP suppressed IL-5 production by human T cell line following T cell receptor (TCR)-stimulation. PGE2 suppressed TCR-induced messenger RNA (mRNA) expression of IL-2, IL-4 and IL-5, as well as proliferation and CD25 expression. 3. Cyclic AMP-mediated suppression of cytokine synthesis, proliferation and CD25 expression in human T cells were attenuated by ionomycin. 4. [cAMP]i was increased by PGE2 and forskolin. PGE2 suppressed the TCR-induced biphasic increase in [Ca2+]i. EMSA revealed that four specific protein-DNA binding complexes related to NF-AT were detected at the IL-5 promoter sequence located from -119 to -90 relative to the transcription initiation site. The slowest migrating complex induced by TCR stimulation was enhanced by PGE2 and further upregulated by ionomycin. Another binding which did not compete with cold AP-1 oligonucleotides, was constitutively present and was unaffected by PGE2 but enhanced by ionomycin. 5. The suppressive effect of cyclic AMP on human IL-5 synthesis is mediated by interference with intracellular Ca2+ mobilization but distinct from the NF-AT-related pathway. PMID- 10385255 TI - Pharmacological analysis of CCK2 receptor antagonists using isolated rat stomach ECL cells. AB - 1. Gastrin stimulates rat stomach ECL cells to secrete histamine and pacreastatin, a chromogranin A (CGA)-derived peptide. The present report describes the effect of nine cholecystokinin2 (CCK2) receptor antagonists and one CCK1 receptor antagonist on the gastrin-evoked secretion of pancreastatin from isolated ECL cells. 2. The CCK2 receptor antagonists comprised three benzodiazepine derivatives L-740,093, YM022 and YF476, one ureidoacetamide compound RP73870, one benzimidazole compound JB 93182, one ureidoindoline compound AG041R and three tryptophan dipeptoids PD 134308 (CI988), PD135158 and PD 136450. The CCK1 receptor antagonist was devazepide. 3. A preparation of well functioning ECL cells (approximately 80% purity) was prepared from rat oxyntic mucosa using counter-flow elutriation. The cells were cultured for 48 h in the presence of 0.1 nM gastrin; they were then washed and incubated with antagonist alone or with various concentrations of antagonist plus 10 nM gastrin (a maximally effective concentration) for 30 min. Gastrin dose-response curves were constructed in the absence or presence of increasing concentrations of antagonist. The amount of pancreastatin secreted was determined by radioimmunoassay. 4. The gastrin-evoked secretion of pancreastatin was inhibited in a dose-dependent manner. YM022, AG041R and YF476 had IC50 values of 0.5, 2.2 and 2.7 nM respectively. L-740,093, JB93182 and RP73870 had IC50 values of 7.8, 9.3 and 9.8 nM, while PD135158, PD136450 and PD134308 had IC50 values of 76, 135 and 145 nM. The CCK1 receptor antagonist devazepide was a poor CCK2 receptor antagonist with an IC50 of about 800 nM. 5. YM022, YF476 and AG041R were chosen for further analysis. YM022 and YF476 shifted the gastrin dose-response curve to the right in a manner suggesting competitive antagonism, while the effects of AG041R could not be explained by simple competitive antagonism. pK(B) values were 11.3 for YM022, 10.8 for YF476 and the apparent pK(B) for AG041R was 10.4. PMID- 10385257 TI - Selective inhibition of inducible nitric oxide synthase prevents ischaemic brain injury. AB - 1. The aim of this study was to investigate the effect of N-(3 (aminomethyl)benzyl)acetamidine (1400W), a selective inhibitor of inducible calcium-independent nitric oxide synthase (iNOS), on the functional and histopathological outcomes of experimental transient focal cerebral ischaemia in rats. 2. Transient ischaemia was produced by the occlusion for 2 h of both the left middle cerebral artery and common carotid artery. Treatments with 1400W (20 mg kg(-1)) or vehicle were started 18 h after occlusion of the arteries and consisted in seven subcutaneous injections at 8 h interval. Ischaemic outcomes and NOS activities (constitutive and calcium-independent NOS) were evaluated 3 days after ischaemia. 3. 1400W significantly reduced ischaemic lesion volume by 31%, and attenuated weight loss and neurological dysfunction. 4. 1400W attenuated the calcium-independent NOS activity in the infarct by 36% without affecting the constitutive NOS activity. 5. These findings suggest that iNOS activation contributes to tissue damage and that selective inhibitors of this isoform may be of interest for the treatment of stroke. PMID- 10385256 TI - Tranilast inhibits protein kinase C-dependent signalling pathway linked to angiogenic activities and gene expression of retinal microcapillary endothelial cells. AB - 1. Tranilast, first developed as an anti-allergic drug, has been reported to inhibit vascular endothelial growth factor (VEGF)-induced angiogenesis and vasopermeability. To further clarify the inhibitory mechanism, we investigated the effects of tranilast on VEGF binding and subsequent intracellular signalling pathway linked to angiogenic activities and gene expression of bovine retinal microcapillary endothelial cells. 2. Tranilast significantly (P<0.01) inhibited VEGF, basic fibroblast growth factor (bFGF), and hypoxia conditioned media induced BREC proliferation in a dose dependent manner with IC50's of 22, 82 and 10 microM, respectively. 3. VEGF-induced migration was also inhibited by tranilast in a dose dependent manner, with IC50 of 18 microM, and complete inhibition was observed at 300 microM (P<0.01). Tranilast suppressed VEGF-induced tube formation in a dose dependent manner with maximum (46%) inhibition observed at 300 microM (P<0.05). 4. Tranilast inhibited phorbol myristate acetate (PMA) dependent stimulation of [3H]-thymidine incorporation and VEGF- and PMA-induced gene expression of integrin alpha v and c-fos in BREC. 5. Tranilast suppressed VEGF- and PMA-stimulated PKC activity in BREC. 6. Tranilast did not affect VEGF binding or VEGF-induced phosphorylation of tyrosine residues of VEGF receptor- and phospholipase Cgamma and their associated proteins. 7. These data suggest that tranilast might prove an effective inhibitor to prevent retinal neovascularization in ischaemic retinal diseases, and that its inhibitory effect might be through suppression of PKC-dependent signal transduction in BREC. PMID- 10385258 TI - Activation of outward K+ currents: effect of VIP in oesophagus. AB - 1. Electrical field stimulations (EFS) of the opossum and canine lower oesophageal sphincters (OLOS and CLOS respectively) and opossum oesophageal body circular muscle (OOBCM) induce non-adrenergic, non-cholinergic (NANC) relaxations of any active tension and NO-mediated hyperpolarization. VIP relaxes the OLOS and CLOS and any tone in OOBCM without major electrophysiological effects. These relaxations are not blocked by NOS inhibitors. Using isolated smooth muscle cells, we tested whether VIP acted through myogenic NO production. 2. Outward currents were similar in OOBCM and OLOS and NO increased them regardless of pipette Ca2+(i), from 50-8000 nM. L-NAME or L-NOARG did not block outward currents in OLOS at 200 nM pipette Ca2+. 3. Outward currents in CLOS cells decreased at 200 nM pipette Ca2+ or less but NO donors still increased them. VIP had no effect on outward currents in cells from OOBCM, OLOS or CLOS under conditions of pipette Ca2+ at which NO donors increased outward K+ currents. 4. We conclude, VIP does not mimic electrophysiological effects of NO donors on isolated cells of OOBCM, OLOS or CLOS. VIP relaxes the OLOS and CLOS and inhibits contraction of OOBCM by a mechanism unrelated to release of myogenic NO or an increase in outward current. 5. Also, the different dependence of outward currents of OOBCM and OLOS on pipette Ca2+ from those of CLOS suggests that different K+ channels are involved and that myogenic NO production contributes to K+ channel activity in CLOS but not in OLOS or OOBCM. PMID- 10385259 TI - Expression of P2Y receptors in cell lines derived from the human lung. AB - 1. Northern blotting experiments have been performed with RNA extracted from several cell lines derived from the human lung in order to detect P2Y1, P2Y2, P2Y4 and P2Y6 mRNA. We have investigated the 1HAEo- and 16HBE14o- epithelial cell lines derived from the airway epithelium, the A549 cell line displaying properties of type II alveolar epithelial cells, the CALU-3 serous cells, the 6CFSMEo- submucosal cells and the HASMSC1 airway smooth muscle cells. We have also evaluated one pancreatic epithelial cell line called CFPAC-1. These experiments revealed that P2Y2 and P2Y6 mRNA are co-expressed in the IHAEo-, 16HBE14o- and A549 epithelial cell lines. The CFPAC-1 pancreatic cell line was strongly positive for the P2Y2 receptor. No signal was obtained for the P2Y1 and P2Y4 receptors. 2. We have then performed RT-PCR experiments with specific oligonucleotides of these last two P2Y receptors with the RNA used for the Northern blotting experiments. P2Y4 mRNA was detected in five cell lines: 1HAEo-, 16HBE14o-, 6CFSMEo-, HASMSC1 and CFPAC-1. P2Y1 mRNA was only detected in the CALU 3 cell line. 3. Inositol trisphosphates assays have identified a response typical of the P2Y2 receptor in the 1HAEo- and the 16HBE14o- airway epithelial cell lines which co-express P2Y2 and P2Y6 mRNA. By contrast, the 6CFSMEo- submucosal cells expressed a UTP-specific response which displayed pharmacological characteristics compatible with the human P2Y4 receptor: in particular, there was no response to UDP or ATP and the UTP effect was totally inhibited by pertussis toxin. PMID- 10385260 TI - Bradykinin down-regulates LPS-induced eosinophil accumulation in the pleural cavity of mice through type 2-kinin receptor activation: a role for prostaglandins. AB - 1. The role of both exogenously administered and endogenously generated bradykinin (BK) on LPS-induced eosinophil accumulation in the mice pleural cavity was investigated by means of treatment with BK selective receptor agonists/antagonists and captopril. 2. Intrathoracic (i.t.) injection of LPS (250 ng cavity(-1)) induced eosinophil influx at 24 h as previously described (Bozza et al., 1993). Pretreatment with the B1 receptor antagonist des-Arg9-[leu-8]BK (0.025 and 0.25 nmol cavity(-1)) showed no effect on this phenomenon, whereas pretreatment with the B2 receptor antagonists, NPC 17731 (0.025 and 0.25 nmol cavity(-1)) or HOE 140 (2.5 nmol cavity(-1)), increased LPS-induced eosinophil influx. Accordingly, pretreatment with captopril at 10 mg kg(-1) i.p., inhibited eosinophil infiltration induced by LPS in the pleural cavity, suggesting that endogenous BK is down-regulating LPS-induced eosinophil accumulation. 3. BK administered at 15 and 25 nmol cavity(-1), i.t. or i.p. also inhibited LPS induced eosinophil accumulation. BK alone had no effect on the basal number of leucocytes in the pleural or peritoneal cavity in doses up to 25 nmol cavity(-1). Nevertheless, when injected at doses of 50 and 100 nmol cavity(-1) BK induced leucocyte influx characterized by neutrophil and eosinophil accumulation at 24 h. 4. Similarly to what was observed with BK, a specific B2 receptor agonist, Tyr8BK, administered at 0.25 nmol cavity(-1) i.p., significantly inhibited the eosinophil influx induced by LPS. 5. The mechanism by which B2 receptor agonists inhibit LPS-induced eosinophil accumulation was investigated by pretreating the animals with indomethacin or a selective cyclooxygenase-2 inhibitor, NS-398. Pretreatment with either indomethacin or NS-398 had no effect on eosinophil influx induced by LPS alone, but those drugs were able to restore the LPS-induced eosinophil influx in Tyr8BK (0.25 nmol cavity(-1)) injected mice. 6. In conclusion, endogenously generated bradykinin seems to modulate, through activation of B2 receptors, eosinphil accumulation induced by LPS via a mechanism dependent on prostanoid synthesis. PMID- 10385261 TI - The effects of verapamil and diltiazem on N-, P- and Q-type calcium channels mediating dopamine release in rat striatum. AB - 1. The putative inhibitory effects of verapamil and diltiazem on neuronal non-L type Ca2+ channels were studied by investigating their effects on either K+- or veratridine-evoked [3H]-dopamine ([3H]-DA) release in rat striatal slices. Involvement of N-, P- and Q-type channels was identified by sensitivity of [3H] DA release to omega-conotoxin GVIA (omega-CTx-GVIA), omega-agatoxin IVA (omega Aga-IVA) and omega-conotoxin MVIIC (omega-CTx-MVIIC), respectively. 2. KCl (50 mM)-evoked [3H]-DA release was abolished in the absence of Ca2+, and was insensitive to dihydropyridines (up to 30 microM). It was significantly blocked by omega-CTx-GVIA (1 microM), omega-Aga-IVA (30 nM) and was confirmed to be abolished by omega-CTx-MVIIC (3 microM), indicating involvement of N-, P- and Q type channel subtypes. 3. Verapamil and diltiazem inhibited K+-evoked [3H]-DA release in a concentration-dependent manner. The inhibitory effects of verapamil or diltiazem (each 30 microM) were fully additive to the effect of omega-CTx-GVIA (1 microM), whereas co-application with omega-Aga-IVA (30 nM) produced similar effects to those of omega-Aga-IVA alone. 4. As shown previously, veratridine evoked [3H]-DA release in Ca2+ containing medium exclusively involves Q-type Ca2+ channels. Here, diltiazem (30 microM) did not inhibit veratridine-evoked [3H]-DA release, whereas verapamil (30 microM) partially inhibited it, indicating possible involvement of Q-type channels in verapamil-induced inhibition. However, verapamil (30 microM) inhibited this release even in the absence of extracellular Ca2+, suggesting that Na+ rather than Q-type Ca2+ channels are involved. 5. Taken together, our results suggest that verapamil can block P- and at higher concentrations possibly N- and Q-type Ca2+ channels linked to [3H]-DA release, whereas diltiazem appears to block P-type Ca2+ channels only. PMID- 10385262 TI - Differential regulation of bradykinin receptor density, intracellular Ca2+, and prostanoid release in skin and foreskin fibroblasts. Effects of cell density and interleukin-1alpha. AB - 1. Bradykinin (BK) receptors, cytosolic Ca2+, and prostanoids were studied in human skin and foreskin fibroblasts. 2. Bmax values of BK receptors were higher in foreskin than in skin fibroblasts, increasing with cell densities in both cell types. IL-1alpha-dependent receptor induction was blocked by cycloheximide. 3. BK stimulated cytosolic Ca2+ elevation was higher in confluent than in non-confluent cultures and larger in foreskin than in skin fibroblasts. Responses were not enhanced after IL-1-alpha-induced up-regulation of BK receptors. 4. Intrinsic prostanoid production was higher in foreskin than in skin fibroblasts at comparable cell densities. In foreskin, but not in skin fibroblasts, BK stimulation increased the release of PGE2 10 fold and that of 6-oxo-PGF1alpha 6-7 fold. 5. Preincubation with IL-1alpha had a marked effect on prostanoid release in foreskin fibroblasts only. Subsequent BK stimulation increased the release of PGE2 and 6-oxo-PGF1alpha 7-10 fold in skin fibroblasts while this increase was only 30% in foreskin fibroblasts. Release of TXA2 reached values up to one third of the other prostanoids. The IL-1alpha induced rise in BK-stimulated PGE2 synthesis was fully abolished by specific inhibition of cyclo-oxygenase 2. 6. IL 1alpha sensitized BK-stimulated prostanoid synthesis and modulated prostanoid patterns differently in fibroblasts from skin and foreskin. The IL-1alpha effects on prostanoid release were not related to BK receptor numbers nor to the BK stimulated Ca2+ signal but appear to be due to induction of prostanoid synthesizing enzymes. Foreskin fibroblasts seem to be unique and significantly different from fibroblasts of other skin locations in respect to their response to inflammation-associated kinins and cytokines. PMID- 10385263 TI - Comparative pharmacology of recombinant human M3 and M5 muscarinic receptors expressed in CHO-K1 cells. AB - 1. Affinity estimates were obtained for several muscarinic antagonists against carbachol-stimulated [3H]-inositol phosphates accumulation in Chinese hamster ovary (CHO-KI) cells stably expressing either human muscarinic M3 or M5 receptor subtypes. The rationale for these studies was to generate a functional antagonist affinity profile for the M5 receptor subtype and compare this with that of the M3 receptor, in order to identify compounds which discriminate between these two subtypes. 2. The rank order of antagonist apparent affinities (pK(B)) at the muscarinic M5 receptor was atropine (8.7) > or =tolterodine (8.6) = 4 diphenylacetoxy-N-methylpiperidine (4-DAMP, 8.6)> darifenacin (7.7) > or =zamifenacin (7.6)>oxybutynin (6.6)= para-fluorohexahydrosiladifenidol (p-F HHSiD, 6.6)>pirenzepine (6.4) > or = methoctramine (6.3)=himbacine (6.3)>AQ-RA 741 (6.1). 3. Antagonist apparent affinities for both receptor subtypes compare well with published binding affinity estimates. No antagonist displayed greater selectivity for the muscarinic M5 subtype over the M3 subtype, but himbacine, AQ RA 741, p-F-HHSiD, darifenacin and oxybutynin displayed between 9- and 60 fold greater selectivity for the muscarinic M3 over the M5 subtype. 4. This study highlights the similarity in pharmacological profiles of M3 and M5 receptor subtypes and identifies five antagonists that may represent useful tools for discriminating between these two subtypes. Collectively, these data show that in the absence of a high affinity M5 selective antagonist, affinity data for a large range of antagonists is critical to define operationally the M5 receptor subtype. PMID- 10385265 TI - Interactions of new and conventional H3-antagonists with non-histaminergic receptors involved in neurogenic and myogenic contractile responses of guinea-pig ileum. AB - 1. Possible effects of new and conventional H3-receptor antagonists towards various non-histaminergic receptors (alpha2-adrenergic, 5-HT3-serotonin, mu opiate, A1-adenosine, M1-and M3-muscarinic) involved in neurogenic and myogenic contractile responses of guinea-pig ileum are investigated. 2. When the isolated ileum was contracted by the 5-HT3 receptor agonist, 2-methyl-5-HT (5 x 10(-7)-8 x 10(-6) M), acetylcholine (1 x 10(-9)-1 x 10(-7) M), KCl (3 x 10(-2) M) or electrical stimulation some of the drugs, included thioperamide and clobenpropit, reduced the contractile response when tested at micromolar concentrations (1-3 x 10(-5) M) (only compound IV exhibited an M3 competitive antagonism with a pK(B) = 5.49 +/- 0.18). 3. Ileal twitch responses to electrical stimulation were dose dependently inhibited by the selective agonists clonidine (3 x 10(-10)-1 x 10(-7) M), dermorphin (1 x 10(-11)-1 x 10(-8) M), R-N6-(2-phenylisopropyl)-adenosine (1 x 10(-9)-3 x 10(-8) M) and McN-A-343 (1 x 10(-7)-1 x 10(-5) M) with different potencies and comparable efficacy (spike amplitude reduction > 85%). All the H3 antagonists under study (up to 1 x 10(-5) M) showed no or minor interactions at the neuronal sites except the compound V which behaved as a weak competitive antagonist at alpha2-adrenoreceptors (pK(B) = 5.96 +/- 0.06). 4. In conclusion, both new and conventional H3-blockers interacted at the enteric neuronal sites here studied with a 1000-30,000 fold lower antagonistic potency than that previously reported for the ileal H3 histamine receptors. Their spasmolytic activity precludes firm conclusions about the non-competitive interaction with 5 HT3 ileal receptor which requires further investigations. PMID- 10385264 TI - Modulation by nitric oxide of spontaneous mechanical activity in rat proximal colon. AB - 1. In order to examine the role of nitric oxide (NO) in the tonic neural inhibition in rat proximal colon, the effects of N(omega)-nitro-L-arginine methyl ester (L-NAME) were studied on the spontaneous contractions of circular muscle (monitored as intraluminal pressure changes) and of longitudinal muscle (detected as isometric tension changes). 2. L-NAME (3 x 10(-6)-3 x 10(-4) M) caused a concentration-dependent increase in the amplitude of circular contractions, without affecting those of longitudinal muscle. This effect was prevented by L arginine (1-5 x 10(-3) M), but not D-arginine. 3. In the presence of tetrodotoxin (10(-6) M), which per se induced increase of the pressure waves, L-NAME (10(-4) M) caused no further effects on the amplitude of the spontaneous contractions. 4. The response to L-NAME (10(-4) M) was unaffected by atropine (10(-6) M), guanethidine (10(-6) M), hexamethonium (up to 3 x 10(-4) M) or alpha-chymotrypsin (up to 5 U ml(-1)). 5. NK2 receptor antagonists, SR 48968 (3 x 10(-6) M) or MEN 10627 (10(-6) M), produced a reduction of the amplitude of the pressure waves but failed to affect the contractile response to L-NAME (10(-4) M). 6. These findings suggest that tonic production of NO from inhibitory neurones influences the degree of contractions of circular muscle. An involvement of an inhibitory peptide as well as disinhibition of cholinergic or NK2-tachykinergic excitatory neurotransmission in the mechanism of NO action can be ruled out. PMID- 10385266 TI - Altered effects of acetylcholine on cyclic AMP and GMP induced changes in O2 consumption of hypertrophic dog cardiac myocytes. AB - 1. We hypothesized that acetylcholine would attenuate the metabolic effect of increasing cAMP and decreasing cGMP on cardiac myocyte O2 consumption (VO2) in dog, and this effect would be altered in left ventricular hypertrophy (LVH) produced by aortic valve placation. 2. Steady-state VO2 of a suspension of ventricular myocytes from control (n = 7) and LVH (n = 6) dogs was measured by Clark O2 electrodes during electrical stimulation (5 ms, 1 Hz, in 2 mm Ca2+). Cyclic AMP and cyclic GMP were determined by radioimmunoassay. Cellular cAMP was increased by forskolin (adenylate cyclase stimulator) and cGMP was decreased by LY83583 (guanylate cyclase inhibitor) both at 10(-7,-6,-5,-4) M with and without 10(-6) M acetylcholine. 3. Baseline cGMP level in LVH (62 +/- 10 fmol 10(-5) myocytes) was significantly greater than that in control (20 +/- 3), although the myocyte VO2 (356 +/- 39 nL O2 min(-1) 10(-5) myocytes) and cAMP levels (3.9 +/- 0.6 nmol 10(5-1) myocytes) were similar to control (312 +/- 23 and 6.9 +/- 3.1). 4. Forskolin increased myocyte cAMP in both control and LVH myocytes and increased VO2 by 51 +/- 13 in control and 91 +/- 65 in LVH myocytes. LY83583 decreased myocyte cGMP levels in control and LVH myocytes and increased VO2 by 128 +/- 57 in control and 43 +/- 26 in LVH myocytes. 5. Acetylcholine altered the cAMP, cGMP, and VO2 levels in control to 2.4 +/- 0.4, 30 +/- 3 and 213 +/- 27 and LVH to 2.5 +/- 0.3, 85 +/- 9 and 261 +/- 32. Acetylcholine attenuated the maximal effects of forskolin on VO2 to 32 +/- 27 in control and 66 +/- 56 in LVH myocytes. Acetylcholine also decreased the maximal effects of LY83583 to 82 +/- 50 in control and 19 +/- 19 in LVH myocytes. 6. The positive metabolic effects of both increases in myocyte cAMP and decreases in cGMP were blunted by acetylcholine. There was a significant increase in myocyte cGMP with forskolin in LVH myocytes. Acetylcholine decreased the increased myocyte VO2 caused by elevated cAMP or decreased cGMP in both control and LVH myocytes, although the absolute decrease in cAMP was reduced and the absolute values of cGMP were higher in LVH myocytes. PMID- 10385267 TI - Nitrergic and purinergic mechanisms and their interactions for relaxation of the rat internal anal sphincter. AB - 1. The NANC neuronal mechanisms for relaxations of the rat internal anal sphincter in response to electrical field stimulation (EFS) were studied in isolated preparations in the presence of atropine (1 microM), propranolol (3 microM) and phentolamine (3 microM). 2. EFS-induced relaxations were abolished by tetrodotoxin (1 microM) and reduced to 64% of control by the guanylate cyclase inhibitor ODQ (1 microM), but were not significantly reduced by the nitric oxide synthase inhibitor L-NAME (100 microM) or oxyhaemoglobin (10 microM). However, in the presence of tubocurarine (10 microM) or apamin (0.1 microM), L-NAME or oxyhaemoglobin greatly reduced or abolished EFS-induced relaxations. 3. The EFS induced relaxations were mimicked by NO (10-100 microM) and by ATP (3-10 mM). The relaxations elicited by these agents were not affected by tetrodotoxin, L-NAME, tubocurarine or apamin. However, ATP-induced relaxations were reduced by the combination of L-NAME with tubocurarine or apamin. 4. Nicotine (10-100 microM) produced concentration-dependent relaxations that were abolished by tubocurarine (10 microM) or hexamethonium (200 microM). After desensitisation to nicotine (100 microM) and in its continued presence, the addition of L-NAME (100 microM) resulted in almost complete abolition of EFS-induced relaxations. 5. It is suggested that tubocurarine, hexamethonium and desensitisation to nicotine have an apamin-like action in the rat internal anal sphincter, the main effect being blockade of a purinergic component of the relaxant transmission process. 6. The findings suggest that both nitrergic and purinergic transmissions are involved in EFS-induced NANC relaxations of the rat internal anal sphincter, and there appears to be a complex interaction between these two pathways of transmission. PMID- 10385268 TI - Involvement of cyclic GMP-dependent mechanism in the nitrergic relaxation of the bovine oesophageal groove. AB - 1. The present study was designed to investigate the mechanisms involved in the relaxations to nitric oxide (NO) of bovine oesophageal groove preparations suspended in organ baths for isometric tension recordings. In preparations treated with guanethidine (10(-5) M) and atropine (10(-7) M) to block adrenergic neurotransmission and muscarinic receptors, respectively, NO released from nitrergic nerves by electrical field stimulation (EFS, 0.5-16 Hz, 1 ms duration, 20 s trains) and exogenously applied as an acidified solution of sodium nitrite (NaNO2, 10(-6)-10(-3) M) caused frequency-and dose-dependent relaxations of noradrenaline (NA, 10(-5) M)-precontracted preparations. 2. Incubation with an inhibitor of NO-stimulated soluble guanylate cyclase, 1H-[1,2,4]oxadiazolo[4,3, a]quinoxalin-1-one (ODQ, 3 x 10(-6) M, for 30 min) did not change the basal tension of oesophageal groove strips but inhibited relaxations to EFS and to exogenous NO. 3. Treatment with iberiotoxin (10(-7) M) and apamin (5 x 10(-7) M), which are blockers of large and small conductance Ca2+-activated K+ channels, respectively, did not modify basal tension or the relaxations induced by EFS and exogenous NO. Incubation with iberiotoxin (10(-7) M) or apamin (5 x 10(-7) M) plus ODQ (3 x 10(-6) M) significantly reduced the relaxations to EFS and exogenous NO. However, in both cases the reductions were similar to the inhibition caused by ODQ alone. The combined addition of charybdotoxin (3 x 10( 8) M) and apamin (5 x 10(-7) M) did not change relaxations to EFS or exogenous NO of the bovine oesophageal groove. 4. The blocker of ATP-sensitive K+ channels, glibenclamide (10(-6) M), had no effect on either resting tension or relaxations induced by both EFS and exogenous NO. Combined treatment with ODQ (3 x 10(-6) M) and glibenclamide (10(-6) M) did not produce additional inhibition compared to ODQ alone. 5. The present results indicate that NO acts as an inhibitory neurotransmitter by relaxing bovine oesophageal groove smooth muscle through a guanylate cyclase-dependent mechanism which does not appear to involve the opening of K+ channels. PMID- 10385269 TI - Blood pressure control in kidney transplant recipients: influence of immunosuppression. AB - 1. Disturbances of the blood pressure regulation, probably due to dysfunction of the autonomic nervous system, are well known complications in chronic renal failure. Haemodialysis and transplantation have been reported to ameliorate nerve dysfunction. 2. In this study, the function of the blood pressure control was investigated in kidney transplant recipients after longtime haemodialysis treated with ciclosporine A and tacrolimus and compared to healthy individuals. To investigate the influence of immunosuppression, the measurements were performed twice, at low and high whole blood concentrations of ciclosporine and tacrolimus. Besides ciclosporine, tacrolimus, azathioprine and prednisolone no other drugs were used in the group of kidney transplant recipients. 3. Kidney transplant recipients (KTR) treated with ciclosporine showed reduced blood pressure and heart rate responses to the cardiovascular stress tests (head-up tilt and cold pressure test) under basal conditions. Two hours after ciclosporine application, the differences in the responses to cardiovascular stress tests between KTR and controls were significantly more pronounced. 4. Patients with tacrolimus immunosuppression showed a similar blood pressure and heart rate response under basal conditions. Two hours after drug application, the blood pressure response following orthostatism and heart rate response to the cold pressure test were significantly higher in tacrolimus treated patients. 5. Our results indicate, that kidney transplant recipients still express an altered function of the blood pressure control. Furthermore, ciclosporine A and tacrolimus seem to contribute to dysfunction of the blood pressure regulation by their own. Tacrolimus immunosuppression does not seem to offer advantages concerning the function of the blood pressure control as compared to ciclosporine A. PMID- 10385270 TI - In vivo demonstration of alpha-adrenoceptor-mediated positive inotropy in pithed rats: evidence that noradrenaline does not stimulate myocardial alpha adrenoceptors. AB - 1. This study examines whether positive inotropy via alpha-adrenoceptors could be observed in vivo in pithed rats. Cardiac contractility was measured as the maximum rate of rise of left ventricular pressure (dP/dt(max)). Heart rate and aortic blood pressure were also recorded. 2. The selective alpha1-adrenoceptor agonists, methoxamine, cirazoline, amidephrine and phenylephrine caused dose related increases in dP/dt(max). This response was progressively reduced by increasing doses of the alpha1-adrenoceptor antagonist prazosin. However, since the concomitant increase in diastolic blood pressure (DBP) was also blocked, the changes in dP/dt(max) may have been a consequence of increased after load. 3. Adrenaline and noradrenaline also increased dP/dt(max), accompanied by pressor responses. Propranolol (1 mg kg(-1)) antagonized the increase in dP/dt(max) in response to noradrenaline, suggesting beta-adrenoceptor involvement, but not that to adrenaline. The additional presence of prazosin (1 mg kg(-1)) further shifted the dose-response curves for both noradrenaline and adrenaline to the right. 4. Analysis of the increases in dP/dt(max) at predetermined increases in DBP by each agonist revealed three groups of regression lines. Adrenaline in the presence of propranolol and the four selective alpha1-adrenoceptor agonists occupied a common central position. Above this group were adrenaline and noradrenaline in the absence of antagonists; their additional effects on contractility were beta adrenoceptor-mediated since the regression lines were lowered by propranolol. Clearly below the main group of agonists was noradrenaline in the presence of propranolol. 5. Thus, for a given increase in DBP, adrenaline (in the presence of beta-blockade) and the alpha1-adrenoceptor agonists exert an additional inotropic effect to noradrenaline (also in the presence of beta-blockade). This is concluded to be an alpha-adrenoceptor-mediated increase in cardiac contractility which is not shared by noradrenaline. PMID- 10385271 TI - Integrated obesity management: bridging the gap between primary and secondary care. PMID- 10385272 TI - Why should obesity be managed? The obese individual's perspective. AB - Weight loss can be achieved using a variety of different methods, alone or in combination, including energy restricted diets, drug treatment and surgical intervention. The difficulty is maintaining weight loss over a prolonged period. Physicians, dietitians and nurses are often pessimistic about their ability to manage obesity. Such negative attitudes, combined with the erroneous belief that obesity is not a serious medical condition, have adversely affected the level of care received by obese patients. Despite this, the family doctor is often approached for help with weight control. Obese patients are generally well informed about diet and weight issues, and are in a good position to critically assess the weight loss advice given by their doctor. The perception of such advice formed part of a questionnaire completed by obese people (346 female/24 male) who successfully reduced their weight while attending a weight loss group. Eighty per cent had previously been advised by their doctor to lose weight, but guidance on how to do this was generally judged to be poor. Only 22% of subjects received positive advice, although 23% of subjects reported that their doctor's advice was indirectly responsible for their weight loss. Patients derived evident health benefits from their weight loss and were generally given a positive response on returning to their doctor. To help patients lose weight, doctors must realise that obesity is a serious chronic medical condition. Ongoing help and support from doctors and other healthcare professionals is a key element in successful long-term weight management. PMID- 10385273 TI - How to tackle the problem early? The role of education in the prevention of obesity. AB - The major issues that confront the clinician in relation to childhood obesity are identifying children at risk, deciding the goal and focus of therapy, and determining how to maintain weight loss. The severity of obesity and the age at which it is present appear to be significant determinants of whether childhood obesity will persist into adulthood. At any age, severe obesity is more likely to persist, and obesity present in adolescents is much more likely to persist than obesity in young children. If a child has obese parents, the risk that their obesity will persist to adulthood increases, though the magnitude of that risk varies with the age of the child. The goals of therapy depend on the child's age and the severity of obesity-related complications. Assessment of the family's readiness to change represents the first focus of therapy. A reduction in time spent watching television, coupled with family involvement and a diet that aims to reduce or eliminate high caloric density foods is the best approach in most cases. Children or adolescents who have an emergent complication of obesity are candidates for aggressive weight reduction such as the protein modified fast. More aggressive therapies, such as drug therapy or gastric bypass surgery, must be considered as experimental in children and adolescents. PMID- 10385274 TI - Who should be educated? Education strategies: could children educate their parents? AB - The International Obesity Task Force (IOTF) concluded that the prevention of weight gain is easier, less expensive and more effective than treating obesity after it has fully developed. The objective of prevention programmes is to reduce the exposure of populations to the environmental causes of obesity. Public health prevention is based on education and behavioural changes aiming at promoting physical activity and a less energy dense diet. Effective management of overweight in children proved to reduce the number who carry their weight problems into adulthood. It has been proposed that school could play an important role in encouraging healthy eating habits. School-based prevention strategies consider the child as the target of the education programme. A complementary approach considers that the child could play an active role in the transmission of the message of prevention. It is the hypothesis of a prospective intervention study in northern France, the 'Fleurbaix-Laventie-Ville-Sante' Study, that nutritional education of children aged 6-12y at school may not only improve their nutritional knowledge but also influence the dietary habits of the family. Preliminary results indicate that the education programme resulted in encouraging changes in dietary habits, mainly a decreased intake of lipid-rich foods in the family. The effects on body weight evolution remain to be evaluated. The study is in progress with a 10-year follow-up. PMID- 10385275 TI - How should the obese patient be managed? Possible approaches to a national obesity management network. AB - There is clearly a need for novel approaches to obesity and its management. This has been addressed by the Czech Society for the Study of Obesity, which established a multi-level obesity management network in response to the increasing prevalence of obesity in the Czech Republic. This network includes obesity management centres attached to major teaching hospitals, combined with input from obesity specialists, other specialists, primary healthcare physicians and weight reduction groups. Such an obesity management system aims to provide appropriate diagnostic and treatment facilities for various degrees of obesity throughout the country. The proposed density of the obesity management network takes into account the limited resources in the healthcare system. The network is designed to overcome the current poor level of understanding of obesity and the lack of time and financial resources which have been the most significant barriers to more involvement of primary care physicians in obesity management. In order to implement this system, a comprehensive education programme on obesity was initiated with postgraduate courses for obesity specialists and with training for counsellors of weight reduction groups. PMID- 10385276 TI - Steps towards the prevention of obesity and associated complications. AB - The problem of obesity is often not recognised. For example, the prevalence of obesity in Sweden is estimated to be 10%, but a study of a county of 414358 inhabitants and the records from 41 primary healthcare centres found that only 949 (3.1%) of patients were registered as obese. This is alarming, since overweight and obesity can be easily identified and the prevention and treatment of obesity is crucial in order to prevent type 2 diabetes. A screening programme in Kisa, a district of southern Sweden, found that 45% of men and 32% of women were overweight (BMI 25-30 kg/m2), while 12% of men and 17% of women were obese (BMI >30 kg/m2). Among people without diagnosed diabetes, a family history of obesity emerged in 1384 subjects; 707 were overweight or obese (BMI >25 kg/m2), with 270 of these having abdominal obesity. Of 212 of these patients who agreed to an oral glucose tolerance test, 16 were found to have type 2 diabetes and 70 impaired glucose tolerance. It is vital that primary healthcare teams become more active in developing co-ordinated programmes of identification, registration and long-term management of overweight and obese people. PMID- 10385277 TI - Cosmetic surgery. Introduction. PMID- 10385278 TI - Cosmetic surgery: surgical tools--psychosocial goals. AB - What determines patients' goals for cosmetic surgery and their satisfaction with the outcome? Historical trends, body image theory, evolutionary biology, and clinical and experimental psychology each contribute answers. The physical changes that patients seek are typically a means to psychosocial goals. Individual objectives vary, but often share an origin in recurrent painful feelings, thoughts, or experiences. Surgical goals include: (1) changes in emotional states or cognitions; (2) improvement in interpersonal relationships; and (3) an altering of reactions of the larger society. Psychological studies of cosmetic surgery patients have been designed primarily to address two fundamental questions: (1) is there a preoperative psychological profile of cosmetic surgery patients; and (2) does cosmetic surgery produce enduring, beneficial psychological change? The use of specialized screening interview questions, and effective collaboration with mental health providers, help a wider range of patients achieve successful surgical outcomes. PMID- 10385279 TI - Topical agents used in association with cosmetic surgery. AB - Effective cosmetic surgery depends on proper preparation of the skin prior to the procedure, excellent wound care, and an appropriate postoperative skin maintenance program. Accomplishing this goal requires a thorough understanding of topical agents. Substances applied to the skin can alter barrier function, permeability, transepidermal water loss, immune response, wound repair, vasostability, collagen deposition, epidermal turnover, and melanin formation, to name a few. Each of these skin characteristics can affect the quality of the end surgical result. This article discusses topical agents used in association with cosmetic surgery. PMID- 10385280 TI - Recent advances in soft tissue augmentation. AB - Soft tissue augmentation has become increasingly important as more individuals seek aesthetic improvement without major surgical procedures. The choice of an appropriate subcutaneous implant, whether solid or injectable, requires a thorough understanding of the materials available. This review of the literature addresses autologous fat and dermis transplantation, autologous and allogeneic human collagen, bovine collagen, acellular dermal allograft, hyaluronic acid derivatives, expanded polytetrafluoroethylene, polymethyl-methacrylate microspheres, and other potential biomaterials. The search for the perfect material to eradicate rhytids, smooth scars, and fill traumatic detects continues. New products appear, sometimes with great fanfare, which fail to fulfill the promise of a better alternative to what we use now. For this reason, an in-depth understanding of implant materials is necessary for any physician performing soft-tissue augmentation procedures. PMID- 10385281 TI - Blepharoplasty. AB - Blepharoplasty is a popular technique for eyelid rejuvenation. Safe and successful blepharoplasty surgery requires a complete understanding of eyelid anatomy, careful preoperative evaluation, and meticulous attention to intraoperative detail. This article discusses the role of blepharoplasty in periorbital rejuvenation, provides the reader with an overview of the anatomy, patient selection, preoperative evaluation, surgical technique, and complications of blepharoplasty surgery. An overview of the educational process through which surgeons can learn to perform blepharoplasty is also presented. PMID- 10385282 TI - Advanced techniques in liposuction. AB - Liposuction is a procedure that has been widely used since the 1980s. The introduction of the tumescent technique in 1987 greatly improved the safety of the procedure. Innovative techniques and equip ment modifications have continued to evolve, broadening the scope of application and improving the results that can be obtained. This article discusses several recent advancements in liposuction techniques. Three-Dimensional Tumescent Liposculpture (William R. Cook, Coronado, CA) was developed to view contiguous areas of the body as a unit rather than to simply suction fat from one particular area. Advanced techniques have improved the ability to liposuction difficult areas, such as the arms, calves, medial thighs, and the male breast. A combination of syringe-suction and machine assisted liposculpture is sometimes used. Tumescent liposculpture has been combined with pulsed CO2 laser surgery to produce a more effective and less invasive treatment for the aging face, neck, and jowls. External ultrasound assisted liposculpture can produce excellent results. PMID- 10385283 TI - Optical hair removal. AB - Traditional methods of hair removal have proven unsatisfactory for many individuals with excessive or unwanted hair. In the last few years, several lasers and xenon flashlamps have been developed that promise to fulfill the need for a practical, safe, and long-lasting method of hair removal. Aggressive marketing of these has contributed to their popularity among patients and physicians. However, significant controversy and confusion surrounds this field. This article provides a detailed explanation of the scientific underpinnings for optical hair removal and explores the advantages and disadvantages of the various devices currently available (Nd:YAG, ruby, alexandrite, diode lasers, and xenon flashlamp). Treatment and safety guidelines are provided to assist the practitioner in the use of these devices. Although the field of optical hair removal is still in its infancy, initial reports of long-term efficacy are encouraging. PMID- 10385284 TI - Keloids and hypertrophic scars: review and treatment strategies. AB - Keloids and hypertrophic scars represent exuberant forms of scar formation that frequently are pruritic, painful, and occasionally form strictures. As well, they may result in significant cosmetic disfigurement. Recent years have seen an increased understanding in the molecular and biological mechanisms of keloidal scar formation, allowing for the development of more specific therapeutic options for these lesions. Despite these developments, keloids and hypertrophic scars remain difficult to manage. Clinical, histopathological, and biochemical features of keloids and hypertrophic scars, as well as treatment guidelines, are discussed. PMID- 10385285 TI - Hair transplantation in women. AB - Female pattern alopecia is most often caused by androgenetic alopecia, and tends to take on one of several classic patterns. Progressive hair loss in women also tends to carry with it significant emotional and social pressures, and advances in hair transplantation techniques have begun to give women increased options for its therapy. Although female pattern androgenetic alopecia is often a diagnosis of exclusion, hair transplantation surgeons are now able to more effectively treat this difficult condition. Careful patient selection is critical to the success of the procedure, as is meticulous technique and surgical planning. PMID- 10385286 TI - A comparison of graft implantation techniques for hair transplantation. AB - Modern hair restoration surgery involves moving hair-bearing grafts from the posterior scalp into areas of hair loss for correction of androgenic alopecia. Over the past decade, this procedure has been refined in multiple ways for the creation of increasingly natural results. The following is a comparison of commonly used graft harvesting, sectioning, and implantation techniques currently in use for hair restoration today. This comparison includes data on total procedure time, graft cutting time, graft insertion time, labor requirements, and cost. PMID- 10385287 TI - A theoretical study of the contrast recovery and variance of MAP reconstructions from PET data. AB - We examine the spatial resolution and variance properties of PET images reconstructed using maximum a posteriori (MAP) or penalized-likelihood methods. Resolution is characterized by the contrast recovery coefficient (CRC) of the local impulse response. Simplified approximate expressions are derived for the local impulse response CRC's and variances for each voxel. Using these results we propose a practical scheme for selecting spatially variant smoothing parameters to optimize lesion detectability through maximization of the local CRC-to-noise ratio in the reconstructed image. PMID- 10385288 TI - Enhancing the multivariate signal of [15O] water PET studies with a new nonlinear neuroanatomical registration algorithm. AB - This paper addresses the problem of neuro-anatomical registration across individuals for functional [15O] water PET activation studies. A new algorithm for three-dimensional (3-D) nonlinear structural registration (warping) of MR scans is presented. The method performs a hierarchically scaled search for a displacement field, maximizing one of several voxel similarity measures derived from the two-dimensional (2-D) histogram of matched image intensities, subject to a regularizer that ensures smoothness of the displacement field. The effect of the nonlinear structural registration is studied when it is computed on anatomical MR scans and applied to coregistered [15O] water PET scans from the same subjects: in this experiment, a study of visually guided saccadic eye movements. The performance of the nonlinear warp is evaluated using multivariate functional signal and noise measures. These measures prove to be useful for comparing different intersubject registration approaches, e.g., affine versus nonlinear. A comparison of 12-parameter affine registration versus non-linear registration demonstrates that the proposed nonlinear method increases the number of voxels retained in the cross-subject mask. We demonstrate that improved structural registration may result in an improved multivariate functional signal to-noise ratio (SNR). Furthermore, registration of PET scans using the 12 parameter affine transformations that align the coregistered MR images does not improve registration, compared to 12-parameter affine alignment of the PET images directly. PMID- 10385289 TI - Generalized likelihood ratio detection for fMRI using complex data. AB - The majority of functional magnetic resonance imaging (fMRI) studies obtain functional information using statistical tests based on the magnitude image reconstructions. Recently, a complex correlation (CC) test was proposed based on the complex image data in order to take advantage of phase information in the signal. However, the CC test ignores additional phase information in the baseline component of the data. In this paper, a new detector for fMRI based on a generalized likelihood ratio test (GLRT) is proposed. The GLRT exploits the fact that the fMRI response signal as well as the baseline component of the data share a common phase. Theoretical analysis and Monte Carlo simulation are used to explore the performance of the new detector. At relatively low signal intensities, the GLRT outperforms both the standard magnitude data test and the CC test. At high signal intensities, the GLRT performs as well as the standard magnitude data test and significantly better than the CC test. PMID- 10385290 TI - Estimation and detection of myocardial tags in MR image without user-defined myocardial contours. AB - Magnetic resonance (MR) tagging has been shown to be a useful technique for noninvasively measuring the deformation of an in vivo heart. An important step in analyzing tagged images is the identification of tag lines in each image of a cine sequence. Most existing tag identification algorithms require user defined myocardial contours. Contour identification, however, is time consuming and requires a considerable amount of user intervention. In this paper, a new method for identifying tag lines, which we call the ML/MAP method, is presented that does not require user defined myocardial contours. The ML/MAP method is composed of three stages. First, a set of candidate tag line centers is estimated across the entire region-of-interest (ROI) with a snake algorithm based on a maximum likelihood (ML) estimate of the tag center. Next, a maximum a posteriori (MAP) hypothesis test is used to detect the candidate tag centers that are actually part of a tag line. Finally, a pruning algorithm is used to remove any detected tag line centers that do not meet a spatio-temporal continuity criterion. The ML/MAP method is demonstrated on data from ten in vivo human hearts. PMID- 10385291 TI - A new approach for nonlinear distortion correction in endoscopic images based on least squares estimation. AB - Images captured with a typical endoscope show spatial distortion, which necessitates distortion correction for subsequent analysis. In this paper, a new methodology based on least squares estimation is proposed to correct the nonlinear distortion in the endoscopic images. A mathematical model based on polynomial mapping is used to map the images from distorted image space onto the corrected image space. The model parameters include the polynomial coefficients, distortion center, and corrected center. The proposed method utilizes a line search approach of global convergence for the iterative procedure to obtain the optimum expansion coefficients. A new technique to find the distortion center of the image based on curvature criterion is presented. A dual-step approach comprising token matching and integrated neighborhood search is also proposed for accurate extraction of the centers of the dots contained in a rectangular grid, used for the model parameter estimation. The model parameters were verified with different grid patterns. The distortion-correction model is applied to several gastrointestinal images and the results are presented. The proposed technique provides high-speed response and forms a key step toward online camera calibration, which is required for accurate quantitative analysis of the images. PMID- 10385292 TI - CT reconstruction by using the MLS-ART technique and the KCD imaging system--I: low-energy X-ray studies. AB - We investigated the use of the kinestatic charge detector (KCD) combined with the multilevel scheme algebraic reconstruction technique (MLS-ART) for X-ray computer tomography (CT) reconstruction. The KCD offers excellent detective quantum efficiency and contrast resolution. These characteristics are especially helpful for applications in which a limited number of projections are used. In addition, the MLS-ART algorithm offers better contrast resolution than does the conventional convolution backprojection (CBP) technique when the number of projections is limited. Here we present images of a Rando-head phantom that was reconstructed by using the KCD and MLS-ART. We also present, for comparison, the images reconstructed by using the CBP technique. The combination of MLS-ART and the KCD yielded satisfactory images after just one or two iterations. PMID- 10385293 TI - Validation of an optical flow method for tag displacement estimation. AB - We present a validation study of an optical-flow method for the rapid estimation of myocardial displacement in magnetic resonance tagged cardiac images. This registration and change visualization (RCV) software uses a hierarchical estimation technique to compute the flow field that describes the warping of an image of one cardiac phase into alignment with the next. This method overcomes the requirement of constant pixel intensity in standard optical-flow methods by preprocessing the input images to reduce any intensity bias which results from the reduction in stripe contrast throughout the cardiac cycle. To validate the method, SPAMM-tagged images were acquired of a silicon gel phantom with simulated rotational motion. The pixel displacement was estimated with the RCV method and the error in pixel tracking was <4% 1000 ms after application of the tags, and after 30 degrees of rotation. An additional study was performed using a SPAMM tagged multiphase slice of a canine left ventricle. The true displacement was determined using a previously validated active contour model (snakes). The error between methods was 6.7% at end systole. The RCV method has the advantage of tracking all pixels in the image in a substantially shorter period than the snakes method. PMID- 10385294 TI - Three-dimensional artifact induced by projection weighting and misalignment. AB - In recent years, the use of computer graphic techniques to produce three dimensional (3-D) and reformatted images from a set of axial computed tomography (CT) images has gained significant interest. In most cases, the CT images are generated with the projection data set weighted prior to reconstruction, to combat motion artifacts, data inconsistency, or redundant data samples. In this paper, we investigate the potential bias introduced to the reconstruction as a result of the interaction of the projection weights and the isocenter misalignment (ISM). We demonstrate that when the weights applied to the conjugate rays are significantly different, bias will result which favors the sample with a higher weight. Although the error is not easily detected in axial CT images, it can be quite visible in 3-D or multiplanar reformatted (MPR) images. In this paper, we first present a theoretical framework to analyze and predict the bias. The theoretical prediction is validated by both computer simulations and phantom experiments. Several schemes to combat this artifact are subsequently presented; and their effectiveness is demonstrated. PMID- 10385295 TI - Credo biotechnology. PMID- 10385296 TI - Complicated is not complex. PMID- 10385297 TI - Biosafety protocol. New confrontations. PMID- 10385298 TI - Biosafety protocol. Boon or threat to international trade in biotechnology? PMID- 10385299 TI - Biosafety protocol. Cynicism and politics dominate UN biotechnology deliberations. PMID- 10385300 TI - Biosafety protocol. "Biosafety" to assure underdevelopment. PMID- 10385301 TI - Icelandic genetics. PMID- 10385302 TI - Are sequences of plasmid DNA used in gene therapy erroneous? PMID- 10385303 TI - GM crops in Europe. PMID- 10385304 TI - LXR needs an elixir to survive. PMID- 10385305 TI - Analysts, firms pour cold water on SNP Consortium. PMID- 10385306 TI - Japan turns to functional genomics. PMID- 10385307 TI - Critics question direction of new investment group. PMID- 10385308 TI - Geron-Roslin alliance aims broadly at degenerative diseases. PMID- 10385309 TI - Value of Discovery is questioned. PMID- 10385310 TI - Bt resistance plan appraised. PMID- 10385311 TI - In silico predictions; in vivo veritas. PMID- 10385312 TI - Delivering tissue regeneration. PMID- 10385313 TI - Double-stranded DNA arrays: next steps in the surface campaign. PMID- 10385314 TI - Nurturing nature: engineering new antibiotics. PMID- 10385315 TI - Enzymes directly evolving toward commercial applications. PMID- 10385316 TI - Can biotechnology move us toward a sustainable society? PMID- 10385317 TI - Hemoglobin-based blood substitutes: oxygen carriers, pressor agents, or oxidants? AB - Hemoglobin-based blood substitutes are being developed as oxygen-carrying agents for the prevention of ischemic tissue damage and hypovolemic (low blood volume) shock. The ability of cell-free hemoglobin blood substitutes to affect vascular tone through the removal of nitric oxide has also prompted an evaluation of their usefulness for maintaining blood pressure in critically ill patients. Before the clinical potential of these substitutes can be fully realized, however, concerns remain as to the intrinsic toxicity of the hemoglobin molecule, particularly the interference of the heme prosthetic group with the tissue oxidant/antioxidant balance. This review provides some insights into the complex redox chemistry of hemoglobin and places an emphasis on how current knowledge may be exploited both to selectively enhance/suppress specific chemical reaction pathway(s) and to ultimately design safer hemoglobin-based therapeutics. PMID- 10385318 TI - DNA delivery from polymer matrices for tissue engineering. AB - We have proposed engineering tissues by the incorporation and sustained release of plasmids encoding tissue-inductive proteins from polymer matrices. Matrices of poly(lactide-co-glycolide) (PLG) were loaded with plasmid, which was subsequently released over a period ranging from days to a month in vitro. Sustained delivery of plasmid DNA from matrices led to the transfection of large numbers of cells. Furthermore, in vivo delivery of a plasmid encoding platelet-derived growth factor enhanced matrix deposition and blood vessel formation in the developing tissue. This contrasts with direct injection of the plasmid, which did not significantly affect tissue formation. This method of DNA delivery may find utility in tissue engineering and gene therapy applications. PMID- 10385319 TI - Generation of tissue-specific and promiscuous HLA ligand databases using DNA microarrays and virtual HLA class II matrices. AB - Most pockets in the human leukocyte antigen-group DR (HLA-DR) groove are shaped by clusters of polymorphic residues and, thus, have distinct chemical and size characteristics in different HLA-DR alleles. Each HLA-DR pocket can be characterized by "pocket profiles," a quantitative representation of the interaction of all natural amino acid residues with a given pocket. In this report we demonstrate that pocket profiles are nearly independent of the remaining HLA-DR cleft. A small database of profiles was sufficient to generate a large number of HLA-DR matrices, representing the majority of human HLA-DR peptide-binding specificity. These virtual matrices were incorporated in software (TEPITOPE) capable of predicting promiscuous HLA class II ligands. This software, in combination with DNA microarray technology, has provided a new tool for the generation of comprehensive databases of candidate promiscuous T-cell epitopes in human disease tissues. First, DNA microarrays are used to reveal genes that are specifically expressed or upregulated in disease tissues. Second, the prediction software enables the scanning of these genes for promiscuous HLA-DR binding sites. In an example, we demonstrate that starting from nearly 20,000 genes, a database of candidate colon cancer-specific and promiscuous T-cell epitopes could be fully populated within a matter of days. Our approach has implications for the development of epitope-based vaccines. PMID- 10385320 TI - Peptidomimetic compounds that inhibit antigen presentation by autoimmune disease associated class II major histocompatibility molecules. AB - We have identified a heptapeptide with high affinity to rheumatoid arthritis associated class II major histocompatibility (MHC) molecules. Using a model of its interaction with the class II binding site, a variety of mimetic substitutions were introduced into the peptide. Several unnatural amino acids and dipeptide mimetics were found to be appropriate substituents and could be combined into compounds with binding affinities comparable to that of the original peptide. Compounds were designed that were several hundred-fold to more than a thousand-fold more potent than the original peptide in inhibiting T-cell responses to processed protein antigens presented by the target MHC molecules. Peptidomimetic compounds of this type could find therapeutic use as MHC-selective antagonists of antigen presentation in the treatment of autoimmune diseases. PMID- 10385321 TI - Improving antibody affinity by mimicking somatic hypermutation in vitro. AB - In vivo affinity maturation of antibodies involves mutation of hot spots in the DNA encoding the variable regions. We have used this information to develop a strategy to improve antibody affinity in vitro using phage display technology. In our experiment with the antimesothelin scFv, SS(scFv), we identified DNA sequences in the variable regions that are naturally prone to hypermutations, selected a few hot spots encoding nonconserved amino acids, and introduced random mutations to make libraries with a size requirement between 10(3) and 10(4) independent clones. Panning of the hot spot libraries yielded several mutants with a 15- to 55-fold increase in affinity compared with a single clone with a fourfold increased affinity from a library in which mutagenesis was done outside the hot spots. The strategy should be generally applicable for the rapid isolation of higher-affinity mutants of Fvs, Fabs, and other recombinant antibodies from antibody phage libraries that are small in size. PMID- 10385322 TI - Quantifying DNA-protein interactions by double-stranded DNA arrays. AB - We have created double-stranded oligonucleotide arrays to perform highly parallel investigations of DNA-protein interactions. Arrays of single-stranded DNA oligonucleotides, synthesized by a combination of photolithography and solid state chemistry, have been used for a variety of applications, including large scale mRNA expression monitoring, genotyping, and sequence-variation analysis. We converted a single-stranded to a double-stranded array by synthesizing a constant sequence at every position on an array and then annealing and enzymatically extending a complementary primer. The efficiency of second-strand synthesis was demonstrated by incorporation of fluorescently labeled dNTPs (2' deoxyribonucleoside 5'-triphosphates) and by terminal transferase addition of a fluorescently labeled ddNTP. The accuracy of second-strand synthesis was demonstrated by digestion of the arrayed double-stranded DNA (dsDNA) on the array with sequence-specific restriction enzymes. We showed dam methylation of dsDNA arrays by digestion with DpnI, which cleaves when its recognition site is methylated. This digestion demonstrated that the dsDNA arrays can be further biochemically modified and that the DNA is accessible for interaction with DNA binding proteins. This dsDNA array approach could be extended to explore the spectrum of sequence-specific protein binding sites in genomes. PMID- 10385323 TI - Integration complexes derived from HIV vectors for rapid assays in vitro. AB - Of three enzymes encoded by HIV-reverse transcriptase, protease, and integrase only the first two have been exploited clinically as inhibitor targets. Efforts to develop inhibitors of purified integrase protein have yielded many compounds, but none with clinical utility. A different source of integration activity for studies in vitro is provided by replication intermediates isolated from HIV infected cells. These preintegration complexes (PICs) can direct integration of the endogenously synthesized viral cDNA into an added target DNA in vitro. Despite their authentic activities, assays of PICs have not been widely used due to technical obstacles, particularly the requirement for handling large amounts of infectious HIV. Here, we describe greatly improved methods for producing PICs using HIV-based vectors that are capable of establishing an integrated provirus but not a spreading infection. We also report the development of a PIC integration assay using DNA-coated microtiter plates, which speeds assays of PIC integration in vitro. We used this method to screen a library of chemicals related to known integrase inhibitors and found a new compound, quinalizarin sulfate, that displayed enhanced activity against PICs. PMID- 10385325 TI - Conversion of Lactococcus lactis from homolactic to homoalanine fermentation through metabolic engineering. AB - We report the engineering of Lactococcus lactis to produce the amino acid L alanine. The primary end product of sugar metabolism in wild-type L. lactis is lactate (homolactic fermentation). The terminal enzymatic reaction (pyruvate + NADH-->L-lactate + NAD+) is performed by L-lactate dehydrogenase (L-LDH). We rerouted the carbon flux toward alanine by expressing the Bacillus sphaericus alanine dehydrogenase (L-AlaDH; pyruvate + NADH + NH4+ -->L-alanine + NAD+ + H2O). Expression of L-AlaDH in an L-LDH-deficient strain permitted production of alanine as the sole end product (homoalanine fermentation). Finally, stereospecific production (>99%) of L-alanine was achieved by disrupting the gene encoding alanine racemase, opening the door to the industrial production of this stereoisomer in food products or bioreactors. PMID- 10385324 TI - Antisense oligonucleotides directed against the viral RNA polymerase gene enhance survival of mice infected with influenza A. AB - We have investigated the ability of antisense phosphorothioate oligonucleotides to enhance the survival of mice infected with influenza A virus. The oligonucleotides were complementary to sequences surrounding the translation initiation codons of the viral PB2 or PA genes (PB2-as or PA-as, respectively) of the influenza A virus RNA polymerases. Intravenous administration of PB2-as in a complex with a cationic liposome, Tfx-10, significantly prolonged the mean survival time in days and increased overall survival rates of mice infected with the influenza A virus. Liposomally encapsulated PB2-as inhibited viral growth in lung tissues and reduced pulmonary consolidations. Liposomally encapsulated PB2 as could be an effective therapeutic agent against influenza A virus. PMID- 10385326 TI - Improved stearate phenotype in transgenic canola expressing a modified acyl-acyl carrier protein thioesterase. AB - The engineering of crops for selected fatty acid production is one of the major goals of plant biotechnology. The Garm FatA1, an acyl-acyl carrier protein (ACP) thioesterase isolated from Garcinia mangostana, generates an elevated stearate (18:0) phenotype in transgenic Brassica plants. By site-directed mutagenesis, we generated seven mutants that showed up to a 13-fold increase in specific enzyme activity toward 18:0-ACP in vitro. The seed-specific expression of mutant S111A/V193A in Brassica plants results in transgenic plants that accumulate 55 68% more stearate than plants expressing the wild-type enzyme. Our results demonstrate that a thioesterase can be engineered to increase specific activity and that its improved function demonstrated in vitro is retained in vivo. PMID- 10385327 TI - Transformation of Aspergillus awamori by Agrobacterium tumefaciens-mediated homologous recombination. AB - Agrobacterium tumefaciens is known to transfer part of its tumor-inducing (Ti) plasmid to the filamentous fungus Aspergillus awamori by illegitimate recombination with the fungal genome. Here, we show that when this Ti DNA shares homology with the A. awamori genome, integration can also occur by homologous recombination. On the basis of this finding, we have developed an efficient method for constructing recombinant mold strains free from bacterial DNA by A. tumefaciens-mediated transformation. Multiple copies of a gene can be integrated rapidly at a predetermined locus in the genome, yielding transformants free of bacterial antibiotic resistance genes or other foreign DNA. Recombinant A. awamori strains were constructed containing up to nine copies of a Fusarium solani pisi cutinase expression cassette integrated in tandem at the pyrG locus. This allowed us to study how mRNA and protein levels are affected by gene copy number, without the influence of chromosomal environmental effects. Cutinase mRNA and protein were maximal with four gene copies, indicating a limitation at the transcriptional level. This transformation system will potentially stimulate market acceptance of derived products by avoiding introduction of bacterial and other foreign DNA into the fungi. PMID- 10385328 TI - Public access to data may be closer than you think. PMID- 10385329 TI - Probe generation by PCR coupled with ligation. PMID- 10385330 TI - Agricultural biotechnology. PMID- 10385331 TI - Gold--a controversial sensitizer. European Environmental and Contact Dermatitis Research Group. AB - Until recently, gold allergy was considered to be extremely rare. Gold has been used and worshipped for thousands of years without any obvious complaints of skin problems, either in those participating in mining and other ways of prospecting, or in those wearing jewellery. When studies on contact allergy to gold sodium thiosulfate were published at the beginning of the 1990s, the allergic nature of the reported positive patch test reactions to gold was questioned. The major argument for such questioning was the lack of demonstrable clinical relevance in most positive reactors. A major reason for the questioning may have been confusion in differentiating between contact allergy and allergic contact dermatitis. To arrive at a diagnosis of allergic contact dermatitis, 3 steps have, in principle, to be fulfilled: (i) establishment of contact allergy; (ii) demonstration of present exposure; (iii) assessment of clinical relevance, i.e., causing or aggravating a contact dermatitis. In this paper, these steps are discussed with regard to gold. With our present knowledge of contact allergy allergic contact dermatitis, we do not recommend including gold sodium thiosulfate in the standard series. It should be applied for scientific purposes and when allergic contact dermatitis from gold is suspected. PMID- 10385332 TI - Isolation and quantification of tuliposides and tulipalins in tulips (Tulipa) by high-performance liquid chromatography. AB - The content of tuliposides and tulipalins were determined in Tulipa species and cultivars by reversed-phase high-performance liquid chromatography (RP-HPLC), using a water:methanol gradient as mobile phase. The compounds were detected by a diode array detector employed at 208 nm. The investigation revealed, in addition to 1- and 6-tuliposide A, tuliposide D and the lactonized aglycones tulipalin A and (-)-tulipalin B, the new tuliposide F and 6-tuliposide B, the latter being a new acyl derivative of the known 1-tuliposide B. All compounds were isolated by preparative RP-HPLC and identified by NMR and mass spectroscopy. The predominant compounds were 6-tuliposide A and B present in amounts up to 1.5% and 1.3% of fresh weight, respectively. 6-Tuliposide A and tulipalin A seem to be the major allergens in tulips, although tuliposide D and F may also contribute to the allergenic properties. Tulipalin A and (-)-tulipalin B occur in intact tulips and are not only produced in response to fungal attack or after excision of the plants. A few species were found to have very low allergen content and a relatively high level of tuliposide B, indicating it should be possible to breed non-allergenic and disease-resistant tulips. PMID- 10385333 TI - Contact allergy to cosmetics: testing with patients' own products. AB - In a 2-year period, 1527 patients with contact dermatitis were investigated in the patch-test clinic. In 531 patients, allergy to cosmetics was suspected from the history and they were tested with their own cosmetic products. 40 (7.5%) (of the 531 patients) had 1 or more positive reactions, 82 (15.4%) had doubtfully positive reaction(s) and 31 (5.8%) had irritant reaction(s). Skin-care products were tested most frequently and were also found to cause most positive, doubtfully positive and irritant reactions, 80% of the patients with positive reactions to their own products had no history of contact dermatitis prior to the presenting attack, and in 92.5% of the cases, the positive reaction was considered relevant or partly relevant. In patients with a positive reaction, ingredients that could be responsible were found in 60% of the cases by testing with the European standard series and a series of common cosmetic ingredients. Fragrance mix and formaldehyde were found to be the ingredients most often responsible and were significantly more frequent in patients with positive reactions to their own products, compared to a control group of eczema patients also seen in the patch-test clinic. PMID- 10385334 TI - Irritancy ranking of anionic detergents using one-time occlusive, repeated occlusive and repeated open tests. AB - Discrepancies between the one-time patch test and the wash test regarding the ranking of irritancy of detergents have been found in the literature. The aim of the present study was to investigate the concordance of irritancy rank order of 4 anionic detergents tested by 3 different exposure methods, namely one-time occlusive, repeated short-time occlusive and repeated short-time open tests. These detergents were sodium cocoyl isethionate (ISE), sodium lauryl sulfate (SLS), soap and disodium lauryl 3-ethoxysulfosuccinate (SUC). The reactions were evaluated by visual scoring and by transepidermal water loss (TEWL) measurement. When scored visually, the rank order in the one-time test was: SOAP > or = SLS > or = ISE > SUC. The other test methods yielded a different order: SLS > ISE > or = SOAP > SUC. A similar rank order was obtained with TEWL measurement for all exposure methods. Generally, the concordance among the different exposure methods was high when evaluated by TEWL. The concordance was lower when evaluation was performed by visual scoring. The present study demonstrates that the choice of exposure model and evaluation method may be important variables influencing the outcome of irritancy testing. It is proposed that the repeated open test is the best way to simulate most in-use situations where the uncovered skin is exposed to detergents. The repeated occlusive test or the one-time patch test may be better to simulate situations in which the skin is occluded after irritation by detergents. PMID- 10385336 TI - Allergic contact dermatitis from natural rubber latex without immediate hypersensitivity. PMID- 10385335 TI - Frequency of delayed-type hypersensitivity to contact allergens in psoriatic patients. PMID- 10385337 TI - Oral allergy syndrome with contact urticaria from cosmetic creams. PMID- 10385338 TI - Contact allergy to corticosteroids in asthma/rhinitis patients. PMID- 10385339 TI - Acute irritant contact dermatitis from propionic acid used in animal feed preservation. PMID- 10385340 TI - Incidence rates of occupational contact urticaria caused by natural rubber latex. PMID- 10385341 TI - Allergic contact dermatitis from povidone-iodine. PMID- 10385342 TI - Allergic contact reactions to colophony presenting as oral disease. PMID- 10385343 TI - Oral lichen planus versus oral lichenoid eruption as a manifestation of contact allergy. PMID- 10385344 TI - Allergic contact dermatitis from colophony and Compositae in a violinist. PMID- 10385345 TI - Contact allergy to methyldibromo glutaronitrile presenting as severe scalp seborrhoeic eczema. PMID- 10385346 TI - Sensitization to cocamidopropylbetaine: an 8-year review. PMID- 10385347 TI - Eyelid dermatitis caused by delayed-type hypersensitivity to natural rubber latex. PMID- 10385348 TI - Drug eruption due to flavoxate hydrochloride. PMID- 10385349 TI - Contact dermatitis from pheniramine maleate in eyedrops. PMID- 10385350 TI - Allergic contact dermatitis due to epirubicin. PMID- 10385351 TI - Pigmented contact dermatitis due to Plathymenia foliosa dust. PMID- 10385352 TI - Piroxicam-beta-cyclodextrin and photosensitivity reactions. PMID- 10385353 TI - Determining causation in epidemiology. AB - Epidemiology uses many methods to identify the causes of disease, although it remains impossible to provide proof that any specific factor is a cause. We are only able to present supporting evidence. We subscribe to the pragmatic view that a factor is indeed a cause if its elimination improves health. We describe a deterministic causal model, where disease develops when the necessary component causes exist in one of several possible constellations of causes, each of which constitutes a sufficient set of component causes. If we accept this concept of disease causation, the notion of multifactorial disease becomes meaningless. We should rather turn our attention to component causes that we can eliminate in order to improve public health. The complex nature of diseases and the fact that it is only possible to present supporting evidence for a causal relationship are some of our reasons for basing the identification of causes on a theoretic model or framework. The purpose is to construct and present a model that is complex enough to formalize basic intuitions concerning cause and effect. Finally, conceptual frameworks provide guidance for the use of multivariable statistical techniques and may assist in the interpretation of the results. PMID- 10385354 TI - Fluoride exposure and dental fluorosis in Newburgh and Kingston, New York: policy implications. AB - OBJECTIVES: This analysis was conducted to determine the changes in the effect of exposure to fluoridation and other sources of fluoride on dental fluorosis in children attending Newburgh and Kingston school districts in New York State. METHODS: Data for this analysis were obtained from two surveys conducted in the 1986 and 1995 school years. Analyses were limited to 3500, 7-14-year-old lifelong residents of a fluoridated or a nonfluoridated community. Dean's classification and DMFS index were used for recording dental fluorosis and caries, respectively. A questionnaire was used to collect fluoride exposure data. Regression procedures were used to estimate the effect of fluoridation, fluoride supplements, and brushing before the age of 2 years on dental fluorosis. RESULTS: Children examined in 1996 were at higher risk for both questionable and very mild to severe dental fluorosis if they received fluoride from water or daily tablet use, or started brushing before the age of 2 years. The increase in risk from 1986 to 1995 was greater for African-American children. CONCLUSION: This analysis showed that the risk of developing dental fluorosis did not decline over time in these communities. Continuous exposure to water fluoridation had an observable effect on dental fluorosis. However, implementation of fluoridation in Newburgh Town did not result in an increase in dental fluorosis prevalence. PMID- 10385356 TI - Extraction of teeth over 5 years in regularly attending adults. AB - OBJECTIVES: This prospective study was conducted to describe the incidence of tooth extraction in a group of regularly attending adults and to assess factors that are predictive of tooth loss. METHODS: Baseline and annual incremental clinical data were obtained from 23 general dental practitioners on a group of their regularly attending, dentate adult patients over a 5-year period. The patients completed a postal questionnaire with questions relating to dental health behaviours, attitudes and knowledge, and social factors. RESULTS: Complete clinical data were obtained from 2799 patients. Four hundred and seventy (17%) patients underwent extractions, 72% of which were posterior teeth. The majority of extractions were for reasons other than caries (79%). Bivariate analyses revealed many significant differences between patients who underwent extractions and those who did not, with respect to the clinical, social, behavioural and attitudinal variables. The logistic regression model for tooth loss included three clinical variables, number of teeth, crowns and sites with recession. Other variables in the final model included the dentist's and patient's prediction of treatment need, having sensitive teeth, having a sweet tooth, living alone and smoking. The sensitivity for the model was 0.57 with specificity 0.72. CONCLUSIONS: This study is unique in its examination of patients and has highlighted that both clinical and other factors are important in predicting who will undergo extractions. Future investigations should assess the consequence of having extractions in terms of health benefit or detriment. PMID- 10385355 TI - Behavioural and emotional problems in children referred to a centre for special dental care. AB - This study was conducted among 203 children (103 boys) referred to a centre for special dental care because of a high level of dental fear. It was undertaken to explore to what extent behavioural and emotional problems co-exist in these children compared with children of a Dutch norm group. The children's parents filled out the parental version of the Child Behaviour Checklist (CBCL), before their first appointment at the centre. The behavioural and emotional problems of the children were assessed by this CBCL, and the mean scores of the children in the study were compared with the mean scores of the norm group. The mean scores on all scales, except on the subscale 'sex problems', of the children with a high level of dental fear proved to be significantly higher than the mean scores of the norm group (P< or =0.001). The results indicated that children referred to a special dental care centre not only suffer from high dental fear but also have problems in several other behavioural and emotional areas. These problems appear to be heterogeneous; they were found in several specific problem areas, both external and internal. PMID- 10385357 TI - A 2-year clinical study of two glass ionomer cements used in the atraumatic restorative treatment (ART) technique. AB - The purpose of the study was to evaluate, in a clinical study over 2 years, the deterioration of two glass ionomer cements used with the atraumatic restorative treatment (ART) technique or approach. Fifty-five Fuji IX and 45 ChemFil Superior restorations were placed randomly in 23 adult patients, mainly in small occlusal preparations in molar teeth. The restorations were placed in a dental hospital by one dentist using the ART technique. Photographs, radiographs and replicas were obtained at baseline and subsequent recalls. Both cements were easy to mix and place, but the radiolucency of ChemFil Superior was a disadvantage. Both cements also showed early high losses of sealant and restorative material. After 2 years, 34.5% of the sealants appeared to be completely lost, with caries recorded in 5.3% of the exposed fissures. In some instances, these small lesions may have been present, but not detected clinically, at the time of sealing. Restoration failures of 7.0% were from wear and fracture of the cements and recurrent caries. Mean cumulative wear was 83.1 microm for Fuji IX and 104.0 microm for ChemFil Superior, which was not statistically significant. The cements became darker after their placement to more closely match the restored teeth, but there were few exact matches. There was no surface staining and only minor marginal discrepancies and staining associated with the restorations. Although the short term clinical performance of the two glass ionomer cements was reasonable, the materials require further improvements in their mechanical properties, to reduce sealant losses and wear. The cements evaluated appear suitable for restricted use only, in posterior teeth. PMID- 10385358 TI - Association of edentulousness with systemic factors in elderly people living at home. AB - OBJECTIVE: To examine the association of edentulousness with systemic factors: age, gender, tobacco-smoking, alcohol intake, body mass index, functioning in daily living, cortical thickness at the mandibular angle, and systemic diseases: bone-fracture (an indicator for osteoporosis), diabetes, thyroid disease, hyperparathyroidism, asthma, heart failure, hypertension. METHODS: The study population comprised 293 elderly subjects, 124 (42%) edentulous and 169 (58%) dentate. The data from clinical and radiographic examinations and structured interviews were analyzed by multiple logistic regression. RESULTS: When edentulousness in both the mandible and the maxilla was considered, history of bone fracture and tobacco-smoking were significantly related to complete edentulousness with odds ratios (OR) of 2.51 (95% CI: 1.47-4.28) and 2.42 (95% CI: 1.32-4.43) respectively, associations independent of age and gender. A similar association was found for the edentulous mandible. In the elderly subjects with an edentulous maxilla, besides the significant factors of history of bone fracture and tobacco-smoking, asthma was also associated with edentulousness at an odds ratio of 10.81 (95% CI: 1.38-84.66), oldest subjects most often being edentulous (OR: 2.23, 95% CI: 1.13-4.39). Diabetes was not related to edentulousness either in the mandible or in the maxilla. CONCLUSIONS: The finding of associations of history of bone fracture, tobacco-smoking, and asthma with edentulousness emphasizes the association of systemic conditions with edentulousness. Advanced age was related to an edentulous maxilla. The relationship between asthma and total tooth loss in the maxilla might suggest a local oral effect of medications used by asthmatic patients. PMID- 10385359 TI - Effect of fluoridated salt intake in infancy: a blind caries and fluorosis study in 8th grade Hungarian pupils. AB - Salt fluoridation is effective at inhibiting caries, but fluorosis prevalence data are deficient. OBJECTIVES: The purpose was to undertake a blind study of caries and tooth mottling in 8th grade school pupils from south-east Hungary who had resided (test) or not resided (control), until November 1985, in a 350 ppm F /kg domestic salt-fluoridated area during their early years of life. METHODS: In Szeged, blind clinical caries and anterior tooth mottling scoring (+10% repeats) of 49 previously salt-fluoridated (mean age 14.14 years) and 59 non-salt fluoridated subjects (mean age 14.08 years) were undertaken by one examiner, in June 1997. In addition, radiographic and photographic recordings were taken. In Glasgow, four dental and two lay staff scored the projected 35 mm colour transparencies (+10% repeats) of each pupil's six upper anterior teeth, for tooth mottling. All clinical, radiographic and photographic data were then analysed. RESULTS: Mean DMFS scores were 9.18 (SD=10.72) for test users and 4.51 (SD=6.24) for control users (P<0.01) and, based on repeat observations, clinical reliability=0.99; X-ray reliability=0.95. Clinically, three test children had fluorosis of 10 teeth, with eight teeth in two controls. Photographic scoring by the clinical examiner gave a 97.2% clinical match, while photographic agreements for all four dentist pairs were 92.5%-97.2%, with lay observers' agreements at 89.8%. For both groups, 10% repeats produced 98.5% agreements. In a sole test case "fluorosis" photographic unanimity was obtained, and non-unanimous "possible fluorosis" was recorded by two to four panel members for only three other test and two control subjects. CONCLUSIONS: No evidence was found that significant anterior tooth fluorosis resulted in subjects exposed previously to 350 ppm F-/kg domestic salt from birth to 2.3-4.8 years of age. However, no caries benefit was demonstrated after the 11.5-year salt fluoridation gap. Caries differences seemed social class-related, city-based controls having less disease than rural test subjects, in spite of an identical F- tablet regimen in all schools from 1987, until subjects were 10 years old. These data emphasise (a) the superiority of sustained community-delivered fluoridation and (b) the need to maintain constant fluoride delivery to tooth surfaces, certainly well beyond 10 years of age. PMID- 10385360 TI - Attachment loss in rural Chinese children over a 3-year period. AB - OBJECTIVES: The periodontal condition of 84 children (9-14 years old, 38 males and 46 females) in a rural area of China was monitored over a 3-year period in order to determine the clinical parameters that act as risk factors for attachment loss. METHODS: Plaque and calculus accumulation (PSS and CI), modified gingival index (MGI), pocket depth (PD) and attachment level (AL) of two quadrants of each child were examined in 1993 and 1996. Following calculation of the mean of each of these parameters and statistical analysis of the change in each of the clinical parameters over the 3-year period, the relationship between attachment loss and clinical parameters was analyzed using multiple regression analysis. RESULTS: At the beginning of this study, the mean values for males and females were 3.66 and 3.58 for PSS, 1.22 and 1.17 for MGI, 1.07 and 0.90 for CI, 2.21 and 2.22 for PD, and both 0.02 for AL respectively. No significant difference between males and females was observed. After 3 years, all except mean PSS had increased significantly. Multiple regression analysis indicated that AL correlated only to age (P<0.01). In 1993, although the AL was > or =1 mm for at least one site in 21 children, none had an AL> or =3 mm. In 1996, the number of children with an AL> or =1 mm had increased to 63, and nine of these children exhibited 3 mm AL (one to six sites per child). No other significant differences were observed between the clinical data of these nine children and those of the other children. CONCLUSION: Although attachment loss tended to increase with age, no clinical parameters correlated with attachment loss in children. PMID- 10385361 TI - Understanding the intention of dentists to provide dental care to HIV+ and AIDS patients. AB - OBJECTIVE: The aim of this study was to understand the intention of dentists to provide dental care to HIV+/AIDS patients. METHODS: A representative sample of 791 dentists from the province of Quebec completed a questionnaire assessing their intention to provide dental care to individuals with HIV+/AIDS as well as their attitudes, perceived social norm, perceived behavioral control, perceived behavioral norm and personal normative belief regarding this behavior. Past experience with providing dental care (habit) to HIV+/AIDS patients, fear of AIDS and socio-demographic characteristics were also assessed. RESULTS: Overall, dentists have a strong intention to provide dental care to HIV+/AIDS patients. Nevertheless, 25% of the respondents expressed a low intention to provide dental care to these patients. The main factors explaining 71% of the variance in intention were perceived behavioral control (beta=0.52, P<0.0001), personal normative belief (beta=0.33, P<0.0001) and habit of treating HIV+/AIDS patients (beta=0.12, P<0.0001). CONCLUSION: To improve the motivation of dentists to treat HIV+/AIDS patients, emphasis should be placed on increasing self-efficacy to cope with the difficulties of providing dental care to HIV+/AIDS patients as well as on the importance of acting in agreement with the Dental Association's Code of Ethics. PMID- 10385362 TI - The provision and outcome of orthodontic services in a Norwegian community: a longitudinal cohort study. AB - In a systematic child dental care system, professionals, consumers, and purchasers have a common interest in ensuring that provision and outcome of orthodontic care fulfill the goals of the service and the public's expectations. The purpose of this prospective study was to examine whether treatment was in fact provided to children with a normative treatment need, and also to assess the outcome of treatment. The baseline data were established for a sample of 83 11 year-olds before decisions about orthodontic treatment had been made, by recording the individuals' orthodontic concern as reported in questionnaires and their occlusal condition according to an index of orthodontic treatment need (NOTI). At re-examination at the age of 16 years, it was also recorded whether the individual had received orthodontic treatment. Most individuals (83%) with a normative need had been treated as well as some individuals without need but who had expressed orthodontic concern. Treatment had resulted in occlusal improvement ('health gain') in most patients, and generally this improvement paralleled a decrease in concern. The goals of the orthodontic service seem to have been fulfilled to the extent that residual treatment need was infrequent. The high treatment rate (63%) and standard may explain why all individuals, both treated and untreated, expressed satisfaction with their dental appearance at 16 years of age. PMID- 10385363 TI - Evaluation of malignancy and the prognosis of esophageal cancer based on an immunohistochemical study (p53, E-cadherin, epidermal growth factor receptor). AB - The subjects in this study consisted of 40 preoperative untreated esophageal squamous cell carcinoma patients. While p53 did not significantly correlate with the clinicopathological factors, E-cadherin significantly correlated with lymphatic invasion, vascular invasion, the depth of invasion, the degree of lymph node metastasis, the histological stage, and the number of lymph node metastases. Epidermal growth factor receptor (EGFR) significantly correlated with age, the depth of invasion, and the number of lymph node metastases. The 5-year cumulative survival rate was 45.7% in the p53-positive cases and 61.9% in the p53-negative cases, with no significant difference, and 87.8% in the E-cadherin-positive cases and 19.1% in the -negative cases, and the difference was significant. The prognosis was significantly poor in EGFR-positive subjects: the 5-year survival rate was 38.6% in EGFR-positive cases and 68% in -negative cases. The 5-year survival rate in E-cadherin-negative, EGFR-positive cases was 0%, while it was 91.7% in the reverse pattern, and this difference was significant. These findings suggest that both E-cadherin and EGFR are important prognostic factors, and a more precise prognosis can thus be obtained by combining them. Such a combined technique may be very useful as an indicator for grading the biological malignancy of esophageal cancer. PMID- 10385364 TI - Impact of splenectomy and immunochemotherapy on survival following gastrectomy for carcinoma: covariate interaction with immunosuppressive acidic protein, a serum marker for the host immune system. Tumor Marker Committee for the Study Group of Immunochemotherapy with PSK for Gastric Cancer. AB - The role of the spleen in tumor immunology is still controversial in that it can either enhance or suppress the antitumor immune response depending on the tumor bearing host. To clarify this biphasic effect of the spleen, a clinical evaluation of splenectomy in conjunction with immunotherapy and the host immune status was performed in gastric cancer patients. The effect of splenectomy and immunotherapy in 253 gastric cancer patients enrolled in a prospective randomized trial (SIP) was analyzed using the Cox's proportional hazards model in terms of the covariate interaction of the preoperative immunosuppressive acidic protein (IAP) level. In patients with high IAP levels (>580 microg/ml) with predicted negative antitumor immune reactions, splenectomy improved the prognosis. In patients with lower IAP values, conversely, the preservation of the spleen and immunotherapy demonstrated a significant benefit to survival. The spleen was shown to have a biphasic activity in terms of its antitumor immune response depending on the IAP level of the patient. The effect of immunotherapy is significantly influenced by the activity of spleen cells. The preoperative IAP level is therefore considered to be a possible indicator for the effectiveness of splenectomy and immunotherapy in curatively resected gastric cancer patients. PMID- 10385365 TI - Co-expression of E-cadherin and alpha-catenin molecules in colorectal cancer. AB - Immunohistochemical staining for epithelial (E)-cadherin and alpha-catenin was performed using frozen sections taken from fresh operative specimens, by the avidin-biotin-peroxidase complex method. Tumors were classified into three types according to the expression modality. Cancer cells with expression at the cell cell boundaries were defined as normal; when the expression was positive, but not concentrated at the cell-cell boundaries, they were defined as cytoplasmic; and when the tumor showed no staining, they were defined as lost. The relationship between these three expression types and the clinicopathological features of colorectal cancer was investigated. In all 50 normal mucosa samples, E-cadherin and alpha-catenin were coexpressed normally. The expression type of E-cadherin and alpha-catenin was normal in 11 and 13 of the cancer tissue specimens, respectively, cytoplasmic in 26 and 29, respectively, and lost in 13 and 8, respectively. Cytoplasmic or lost expression was observed in cancer demonstrating an advanced clinical stage (E-cadherin, P = 0.0065; alpha-catenin, P = 0.0069), advanced tumor penetration (P = 0.0003, P = 0.0001), undifferentiated tumor histology (P = 0.0196, P = 0.0343), widespread lymph node involvement (P = 0.0204, P = 0.0340), and liver metastasis (P = 0.0063, P = 0.0299). In conclusion, the expression type of E-cadherin is significantly correlated to that of alpha-catenin, and the loss of their expression indicates the metastatic potentiality of colorectal cancer. PMID- 10385366 TI - Establishment and characterization of human pancreatic carcinoma lines with a high metastatic potential in the liver of nude mice. AB - To investigate of human pancreatic cancer metastasis to the liver, a pancreatic carcinoma line, HPC-3, was injected into the spleens of nude mice. The cells from a few liver metastatic foci of the mice injected with HPC-3 were expanded in vitro and subsequently injected into the spleens of nude mice. By repeating these procedures, we were able to obtain a cell line, designated HPC-3H4. The mice were observed to have liver metastasis in 6 of 6 (100%) cases injected with HPC-3H4, whereas the rate was 0% at 3 weeks after the intrasplenic injection of HPC-3. The tumorigenicity of HPC-3H4 was more rapid than that of HPC-3. The motile activity of HPC-3H4 was also stronger than that of HPC-3, and the adhesion to the extracellular matrix of HPC-3H4 was stronger than that of HPC-3. We also analyzed the cell surface expression of the metastasis-related adhesion molecules. As a result, no substantial changes were observed in the expression level of adhesion molecules. These results suggest that HPC-3H4 is useful for studies aimed at the prevention of liver metastasis. PMID- 10385367 TI - The mechanism of hepatic graft protection against reperfusion injury by prostaglandin E1. AB - The purpose of this study was to evaluate the effect of prostaglandin E1 (PGE1) on protecting against hepatic endothelial cell damage and increasing graft viability after cold preservation and reperfusion, using an isolated perfused rat liver (IPRL) model. The grafts were divided into three groups, according to the cold preservation time and PGE1 administration, namely: 4h preservation (group 1, n = 9), 6h preservation (group 2, n = 9), and 6h preservation followed by PGE1 infusion (group 3, n = 9). After cold storage, the grafts were put on the recirculating IPRL system, then reperfused for 120 min at 37 degrees C with oxygenated Krebs-Henseleit buffer containing hyaluronic acid (HA). To examine the function of the sinusoidal endothelial cells and hepatocytes, serial measurements of HA, tumor necrosis factor-alpha (TNFalpha), thromboxane B2 (TXB2), acid phosphatase, and conventional parameters in the perfusate were made. After perfusion, the trypan blue exclusion test was performed to assess the presence of any microscopic sinusoidal lining cell damage. In group 3, the bile output and HA clearance were significantly greater, while glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, TNFalpha, TXB2, and acid phosphatase in the perfusate were significantly lower than in group 2. Histologically, less endothelial cell damage and hepatocyte damage than in group 2 was also confirmed. These results therefore suggest that the improvement of hepatic graft viability by PGE1 administration is mainly due to sinusoidal endothelial cell protection. PMID- 10385368 TI - Recombinant retrovirus vectors for the expression of MHC class II heterodimers. AB - Class II antigens are critical in determining the fate of vascularized allografts across major histocompatibility differences. We have recently developed a new approach to induce transplantation tolerance in miniature swine by creating MHC class II antigen "molecular chimerism" in bone marrow cells of potential recipients through retrovirus-mediated gene transfer. As part of this project, the ability of a recombinant double-expression vector (ZQ32N) to express MHC class II DQA and DQB was investigated. Flow cytometry analyses of ZQ32N transfected virus-producer cells demonstrated the cell surface expression of DQa/DQb heterodimers, thus suggesting a correct transcription, translation, and transport of the swine polypeptides to the cell surface. The analyses of RNA isolated from virus particles produced from ZQ32N transfected virus-producer cells indicated the DQ sequences to be correctly packaged. However, the DQ negative cells transduced with the ZQ32N retrovirus did not show any DQ retrovirus surface expression. Southern and Northern blot analyses of ZQ32N transfected and transduced cells strongly suggested DNA rearrangements and deletions which could account for transgene expression loss. An analysis of transduced cell genomes suggested DNA recombinations targeted to homologous sequences within the recombinant provirus. The implications of the sequence instability in designing vectors for gene therapy of organ transplantation are discussed. PMID- 10385369 TI - Lymphoepithelioma-like esophageal carcinoma: report of a case. AB - We herein report the rare case of a patient suffering from lymphoepithelioma-like poorly differentiated squamous cell carcinoma of the esophagus. The patient was a 74-year-old woman in whom an esophageal tumor was found during an operation for thyroid cancer. After performing a subtotal thyroidectomy and cervical esophagectomy, esophageal reconstruction was performed using a free jejunal graft. Based on the results of the pathological examination, the esophageal tumor was diagnosed to be primary lymphoepithelioma-like esophageal cancer, not metastasis of either unknown nasopharyngeal cancer or thyroid cancer. Since surgery, she has survived postoperatively for more than 4 years with no evidence of recurrent disease. PMID- 10385370 TI - Esophageal carcinoma showing a long stricture due to prominent lymphatic permeation: report of a case. AB - Some esophageal diseases such as carcinoma, esophagitis, and collagen diseases have often been reported to show a diffusely thickened esophageal wall in the roentogenogram findings. In the current report, a preoperative upper gastrointestinal series and an endoscopic examination showed a diffusely infiltrative type carcinoma, but other examinations did not suggest any diseases such as esophagitis or collagen diseases which might cause a thickening of the esophageal wall or a constriction of the esophagus. A postoperative histological examination revealed the primary carcinoma to remain only within the mucosal layer, while a large degree of lymphatic vessel permeation reached the adventitia over a wide area. An extraordinary degree of lymphatic permeation spread through the esophageal wall, and stromal fibrosis developed as a result of such lymphatic permeation. These histological phenomena might thus have led to the macroscopic appearance of infiltrative type esophageal carcinoma. PMID- 10385371 TI - Gastric cancer with sarcoid reactions in the regional lymph nodes, the stomach wall, and the splenic parenchyma: report of a case. AB - A 66-year-old man was referred to our institute for investigation of heartburn and epigastralgia. Endoscopic examination demonstrated a type 4' gastric cancer which occupied the whole stomach. At laparotomy, multiple small nodules were found in the spleen which were diagnosed as metastases of the gastric cancer. Thus, total gastrectomy with distal pancreatectomy, splenectomy, cholecystectomy, and left adrenalectomy, combined with D4 lymph node dissection, was performed. Microscopic examination of the tumor revealed tubular and mucinous adenocarcinoma which invaded the muscularis propria. Sarcoid reactions were observed in the submucosa adjacent to the carcinoma tissue. Only one lymph node from station no. 8a demonstrated tumor metastasis, while those from station nos. 1, 2, 7, 8, 9, 10, 11, 13, and 16 revealed sarcoid reactions without tumor metastases. Subsequently, the multiple small nodules that had been presumed to be splenic metastases at laparotomy were found to be sarcoid reactions similar to those seen in the submucosa and regional lymph nodes. Since no skin or ocular lesions indicative of systemic sarcoidosis were seen in this patient, a diagnosis of advanced gastric cancer associated with sarcoid reactions was established. To our knowledge, there have been no previous reports regarding an association between sarcoid reactions in the spleen and gastric cancer. PMID- 10385372 TI - Laparoscopic surgery for blind pouch syndrome following Roux-en Y gastrojejunostomy: report of a case. AB - We report herein the case of a 59-year-old man in whom blind pouch syndrome was successfully treated by laparoscopic surgery. The patient had undergone distal gastrectomy and Roux-en Y gastrojejunostomy for a peptic ulcer 35 years previously, and had been suffering from watery diarrhea, anemia, weight loss, and pain in the left upper quadrant of his abdomen for several years. Long-term insufficient oral intake and the malabsorption of nutrients had resulted in severe emaciation. Gastrointestinal contrast study revealed a large blind pouch, 30 x 23cm in diameter, draining into the gastrojejunostomy. Laparoscopic resection of the blind pouch was performed. Despite the presence of dense intraabdominal adhesions, we identified the blind pouch with the help of tattoo marks that had been made at the neck of the pouch preoperatively. After thoroughly dissecting the adhesions around the pouch, we resected the pouch at the neck. The patient had an uneventful postoperative course. This case report demonstrates that large blind pouches such as this may be effectively treated using laparoscopic surgery. PMID- 10385373 TI - Mucosa-associated lymphoid tissue lymphoma of the duodenum forming multiple polypoid lesions: report of a case. AB - We report herein the case of a patient found to have mucosa-associated lymphoid tissue (MALT) lymphoma of the duodenum forming multiple polypoid lesions. Endoscopic examination revealed multiple small nodules with a yellow-white, rough surface in the duodenal bulb. Histopathological and immunological findings subsequently suggested low-grade B-cell MALT lymphoma. Cytologically, MALT lymphoma is similar to multiple lymphomatous polyposis (MLP); however, this case, which involved multiple polypoid lesions, was confirmed not to be MLP. PMID- 10385374 TI - Intrahepatic cholangiocarcinoma with extensive sarcomatous change: report of a case. AB - A 77-year-old woman was admitted to our hospital with severe upper abdominal pain. Ultrasonography showed a well-defined hypoechoic mass with heterogeneity in the left lobe of the liver, and computed tomography demonstrated a low-density mass with enhanced peripheral areas. Magnetic resonance imaging revealed a mass with iso- to low signal intensity on T1-weighted images (WI) and heterogeneous high and low signal intensity on T2 WI. The tumor was found to be hypovascular by angiography. During 5 months of observation, the tumor increased in size, which strongly suggested malignancy. A laparotomy was performed under the provisional diagnosis of a neoplasm other than hepatocellular carcinoma, revealing that the hepatic mass had invaded the gastric wall. Therefore, a left hepatic lobectomy with dissection of the lymph nodes and hemigastrectomy was carried out. Histologically, the tumor was found to be composed of a large amount of sarcomatous elements and a small amount of adenocarcinomatous elements, both of which were partly intermingled. Immunohistochemically, the sarcomatous element demonstrated the features of malignant fibrous histiocytoma (MFH). Thus, a diagnosis of intrahepatic cholangiocarcinoma with MFH-like sarcomatous change was confirmed. PMID- 10385376 TI - Primary torsion of the omentum in a 6-year-old boy: report of a case. AB - In the absence of any pathological condition in the pelvis of children who present with acute abdomen, the observation of serosanguineous intraabdominal fluid should prompt the surgeon to investigate the omentum. A 6-year-old boy who was admitted with a clinical picture of acute appendicitis, and was later diagnosed during a laparotomy to have primary torsion of the omentum, is reported. Obesity, exercise, and the presence of a bifid omentum were confirmed to be the predisposing factors. PMID- 10385375 TI - Posttraumatic intestinal stenosis presenting as a perforation: report of a case. AB - A 78-year-old woman was admitted to the hospital after falling into a ditch approximately 1 m deep and sustaining a blunt abdominal trauma with a left femur fracture. On the tenth day after admission, symptoms of a small bowel obstruction occurred. A nasogastric tube was inserted, and the symptoms thus improved. She sometimes complained of abdominal pain during the 12 months after the fall, but recovered with conservative management. The next year, she was readmitted to the hospital for a pin extraction of the left femur bone. During this admission, 15 months since her admission after her fall, she again complained of abdominal pain. Abdominal pain increased with a muscular defense, and abdominal X-rays revealed free air. She was referred to our hospital with a diagnosis of perforative peritonitis, and emergency surgery was performed. Upon laparotomy, circumferential stenoses of the small bowel were recognized in the proximal segments about 40cm, 80cm, and 100cm from the ileocecal region. In addition, a perforation and prominent dilatation of the bowel segment was observed just proximal to the stenosis about 100cm from the ileocecal region. She underwent a small intestinal resection at two sites. There were no findings of an intestinal specific ulcer, such as Crohn's disease, intestinal tuberculosis, or malignancy, based on the results of a histopathological examination. PMID- 10385377 TI - Successful resection of endotracheal papillary adenocarcinoma by endoscopic electrosurgery using a new snare: report of a case. AB - We report herein the rare case of a patient with endotracheal papillary adenocarcinoma of the goblet cell type. The tumor existed in the upper trachea, obstructing 95% of the airway, and prompt palliation was required. Endotracheal polypectomy was successfully performed in this patient using electrosurgery with a new snare designed for the respiratory tract. The procedure took only a few minutes and the dyspnea was relieved promptly without any complications. The alternative techniques for palliation of bronchial stenosis are also discussed. PMID- 10385378 TI - Extended survival of a porcine mitral bioprosthesis for 23 years: report of a case. AB - Limited durability is the major drawback of bioprosthetic valves, few of which survive for as long as 20 years. We report herein the case of a patient we recently encountered in whom a bioprothesis lasted for 23 years. To our knowledge, this is only the second case of such long survival. The patient was a 56-year-old man who was urgently admitted to our hospital with acute mitral regurgitation, 23 years after undergoing mitral valve replacement with a porcine bioprosthesis. Acute leaflet tears were found to be the cause of the mitral incompetence and the xenograft was successfully replaced with a mechanical valve. We believe that when reoperation is thought to carry a low risk, prophylactic surgery might be justified, even in patients without symptoms. PMID- 10385379 TI - Atherosclerotic superficial temporal artery aneurysm: report of a case. AB - We herein report a rare case of an atherosclerotic superficial temporal artery (STA) aneurysm. The patient was a 34-year-old Japanese man. He noticed a throbbing swelling just in front of his right ear, which had slowly increased in size. There was no history of trauma. Digital subtraction angiography and ultrasound revealed an aneurysm measuring 2cm in diameter fed by STA. A ligation and resection were performed under local anesthesia. The recovery period was uneventful. A microscopic examination revealed a moderate degree of atherosclerosis. We found 48 cases of STA aneurysm in the Japanese literature from 1969 to 1997, including our case. Thirty-six of these cases (75.0%) were considered to be traumatic in origin. Three cases (6.2%) occurred after a craniotomy. To date, only nine reported cases (18.8%) have been attributed to a spontaneous or congenital etiology. PMID- 10385380 TI - The association of cigarette smoking with a high risk of recurrence after ileocolonic resection for ileocecal Crohn's disease. AB - The purpose of this retrospective study was to examine the influence of cigarette smoking on the recurrence of Crohn's disease after resection. Between 1975 and 1990, 141 patients underwent primary ileocolonic resection for ileocecal Crohn's disease, 79 of whom were nonsmokers and 62 of whom were smokers at the time of their operation. The 5- and 10-year cumulative recurrence-free rates were 65% and 45% for the smokers, and 81% and 64% for the nonsmokers, the former values being significantly lower than the latter values (P = 0.007). The smokers were further divided into two subgroups according to the number of cigarettes smoked per day; as mild smokers who smoked fewer than 15 per day (n = 31) and heavy smokers who smoked 15 or more per day (n = 31). The cumulative recurrence-free rate was lower in the heavy smokers compared with the mild smokers. These findings strongly suggest that smoking is associated with a high recurrence rate after ileocolonic resection for ileocecal Crohn's disease. PMID- 10385382 TI - Extension of McBurney's incision: old standards versus new options. PMID- 10385381 TI - A new simple technique for performing intraoperative endoscopic resection of small-bowel polyps in patients with Peutz-Jeghers syndrome. AB - We describe herein a simple method for performing intraoperative endoscopic resection of small-bowel polyps associated with Peutz-Jeghers syndrome, using a corrugated anesthetic tube. A 34-year-old man with Peutz-Jeghers syndrome underwent emergency surgery for an ileo-ileo-colic intussusception. A sterile corrugated anesthetic tube was inserted into the small-bowel, proximal to the affected lesion. The small bowel was then telescoped sequentially over the tube using a pleating technique. Consequently, a colonoscope inserted through the tube was easily able to reach the duodeno-jejunal junction, and ten small-bowel polyps were removed using a wire snare and electrocauterization. All resected specimens were washed out by the instillation of saline through a nasogastric tube, then collected on gauze placed near the outlet of the tube. Our technique has the following merits: it is feasible even in emergency surgery; it prevents contamination of the surgical field; and it facilitates the easy collection of polypectomized specimens. PMID- 10385383 TI - Nutrition and fat cell differentiation. PMID- 10385384 TI - Modulation of CCAAT/enhancer-binding protein-alpha gene expression by metabolic signals in rodent adipocytes. AB - The transcription factor CCAAT/enhancer-binding protein-alpha (C/EBPalpha) is a positive modulator of transcription for several adipocyte-specific genes that play a role in energy metabolism. However, there is little information available regarding the regulation of its expression by metabolic signals. Exposure to insulin for 5-24 h attenuated C/EBPalpha expression when 3T3-L1 adipocytes were incubated in 24 mM glucose, but not in 5.7 mM glucose. Nuclear run-on transcription assays indicated a transcriptional repression of C/EBPalpha gene, but not that of C/EBPbeta. Glucosamine, a product of the hexosamine pathway, in the presence of low glucose mimicked high glucose's ability to reduce C/EBPalpha messenger RNA expression in insulin-treated cells. Similar results were obtained with xylitol, an activator of the pentose phosphate pathway. There was no correlation between the accumulation of hexosamine pathway metabolites (e.g. UDP N-acetylhexosamines) and/or changes in intracellular protein glycosylation with the ability of high glucose, glucosamine, or xylitol to down-regulate C/EBPalpha gene expression. None of these treatments caused a reduction in intracellular ATP levels. Stable transfection of 3T3-L1 cells with the 5'-flanking 468-bp sequence of the mouse C/EBPalpha gene fused to luciferase demonstrated that promoter activity was also reduced by these nutrients. Of interest, treatment of rats with glucose or glucosamine led to a reduction in C/EBPalpha messenger RNA levels in epididymal, but not omental, fat. Taken together, these results suggest that metabolic signals serve to down-regulate C/EBPalpha expression both in vitro and in vivo. PMID- 10385385 TI - Mammary type I deiodinase is dependent on the suckling stimulus: differential role of norepinephrine and prolactin. AB - Mammary deiodinase type I (M-D1) is present only during lactation and exhibits a clear direct correlation with lactation intensity (size of litters). The present work shows that M-D1 is suckling dependent and that intervals between suckling periods no longer than 12 h are essential to maintain this activity. Moreover, we find that with only 15 min of resuckling in 12-h nonsuckled mothers, the 50% decrease in both M-D1 messenger RNA and enzymatic activity could be restored to control values. This restorative effect by suckling may involve pre- and posttranscriptional mechanisms in which norepinephrine and PRL play important roles. Norepinephrine elicits a potent stimulatory effect on M-D1 messenger RNA and enzyme activities, whereas PRL only increases M-D 1 activity and may modulate the enzyme response to norepinephrine. Oxytocin and GH had no effect. These data suggest that the adrenergic nervous system and PRL could directly participate in mammary energetic expenditure, regulating the local T3 supply. PMID- 10385386 TI - Characterization of immortalized osteoclast precursors developed from mice transgenic for both bcl-X(L) and simian virus 40 large T antigen. AB - We recently developed an immortalized osteoclast (OCL) precursor cell line that forms large numbers of OCLs. This cell line was derived from mice doubly transgenic for bcl-X(L) and large T antigen that was targeted to cells in the OCL lineage (bcl-X(L)/Tag cells). We have now characterized these cells in terms of their surface and enzymatic phenotype, responsiveness to osteotropic factors, and differentiation potential. The bcl-X(L)/Tag cells expressed interleukin-1 receptors 1 and 2, gelatinase B (MMP9), as well as Mac-1, CD16/CD32 (Fcgamma receptors), CD45.2 (common leukocyte marker), CD86 (costimulatory molecule expressed on B cells, follicular dendritic cells, and thymic epithelium), major histocompatibility complex I, and nonspecific esterase when cocultured with MC3T3E1 cells. However, they did not express the antigens for F4/80 (mature macrophage/dendritic cell marker) by immunostaining. Treatment of bcl-X(L)/Tag cells, cocultured with MC3T3E 1 cells, with the combination of 1,25 dihydroxyvitamin D3 and dexamethasone induced high levels of OCL formation. The bcl-X(L)/Tag cells formed large numbers of OCLs when cultured with RANK ligand and macrophage colony-stimulating factor in the absence of feeder cells. In the absence of RANK ligand and a feeder cell layer, 100% of the cells differentiated into F4/80-positive cells. However, neither PTH nor PTH-related protein enhanced OCL formation by bcl-X(L)/Tag cells even when they were cocultured with primary osteoblasts, suggesting that they differ from primary mouse bone marrow cells in their responsiveness to PTH/PTH-related protein. Thus, bcl-X(L)/Tag cells have many of the properties of primary mouse OCL precursors and should be very useful for studies of OCL differentiation and divergence of OCL precursors from the macrophage lineage. PMID- 10385387 TI - Effects of L-arginine in rat adrenal cells: involvement of nitric oxide synthase. AB - The effects of L-arginine on corticosterone production, cGMP, and nitrite levels were examined in zona fasciculata adrenal cells. L-Arginine significantly decreased both basal and ACTH-stimulated corticosterone production. This effect was still evident when steroidogenesis was induced by 8-bromo-cAMP and 22(R) hydroxycholesterol, but not in the presence of exogenously added pregnenolone. L Arginine increased cGMP and nitrite levels,; these effects were blocked by the nitric oxide synthase inhibitor, N(G)-nitro-L-arginine methyl-ester. Transport of L-[3H]arginine was rapid, saturable, and monophasic, with an apparent Km of 163+/ 14 microM and a maximum velocity of 53+/-6 pmol/min x 10(5) cells. The basic amino acids L-lysine and L-ornithine, but not D-arginine or the nitric oxide synthase inhibitors N(G)-nitro-L-arginine methyl-ester and N(G)-nitro-L-arginine, impaired L-arginine uptake. Taken together, these results suggest that steroidogenesis in zona fasciculata adrenal cells may be negatively modulated by L-arginine-derived nitric oxide. PMID- 10385388 TI - Expression of peroxisome proliferator-activated receptor alpha messenger ribonucleic acid and protein in human and rat testis. AB - Peroxisome proliferator-activated receptor a (PPARalpha), a member of the steroid hormone receptor superfamily, has been linked to lipid homeostasis and tumorigenesis in tissues with high expression of receptor protein. On the other hand, the role of PPARalpha in tissues with a lower expression is not well known. Here we demonstrate the localization of PPARalpha messenger RNA (mRNA) and protein in developing and adult rat testis. Additionally, we demonstrate the expression of PPARalpha protein in adult human testis. Our experiments with Northern analysis, in situ hybridization and immunocytochemistry reveal a complex distribution of PPARalpha in tubular and interstitial cells of both adult and developing rat testis. The overall expression is rather low but may be modified by exogenous or endogenous stimuli. An up-regulation of PPARalpha mRNA could be observed after stimulation with FSH. In the developing rat testis, a clear expression of PPARalpha mRNA was present from the first days after birth. Additionally, PPARalpha mRNA and protein increased toward adulthood. In adult human testis PPARalpha immunoreactivity (IR) was present in interstitial Leydig cells and tubular cells. In the seminiferous epithelium of adult human testis the expression of PPARalpha-IR could be seen in meiotic spermatocytes, spermatids and myoid peritubular cells. The findings of our study suggest that PPARalpha may be involved in the regulation of growth and differentiation of tubular and interstitial cells in rat and human testis. PMID- 10385389 TI - Estrogen receptor regulation of the Na+/H+ exchange regulatory factor. AB - To better understand the actions of estrogens and antiestrogens in estrogen target cells, we have searched for estrogen-regulated genes in human breast cancer cells, in which the number of genes known to be directly activated by estrogen is quite small. Using differential display RNA methods, we have identified the human homolog of the Na+ -H+ exchanger regulatory factor (NHE-RF), an approximately 50-kDa protein that is also an ezrin-radixin-moesin-binding phosphoprotein, as being under rapid and direct regulation by estrogen in estrogen receptor (ER)-containing breast cancer cells. Stimulation by estrogen of NHE-RF RNA is rapid, being near maximal (approximately 6-fold) by 1 h, and is not blocked by cycloheximide, indicating that it is a primary response. Stimulation is selective for estrogen ligands, with no stimulation by other classes of steroid hormones, and stimulation by estrogen is suppressed by the antiestrogens tamoxifen and ICI 182,780. Induction is shown to require an active ER through several approaches, including the use of ER-negative breast cancer cells containing a stably integrated ER. NHE-RF protein levels, monitored using antibodies specific for this protein, increase after estrogen and reach maximal levels at 24-48 h. Interestingly, NHE-RF is a PDZ domain-containing protein that is enriched in polarized epithelia, where it is known to be localized in microvilli. Among various human tissues we have examined, we found that NHE-RF is expressed at a fairly high level in mammary tissue. NHE-RF regulates protein kinase A inhibition of the Na+ -H+ exchanger and may serve as a scaffold adaptor protein that contributes to the specificity of signal transduction events. Our findings suggest that the early, known effects of estrogen on cell cytoarchitecture (e.g. increasing microvilli on breast cancer cells) and on some cell signaling pathways (e.g. those involving cAMP) may involve rapid estrogen mediated changes in the production of NHE-RF. PMID- 10385390 TI - Transient expression of c-erbAbeta1 messenger ribonucleic acid and beta1 thyroid hormone receptor early in adipogenesis of Ob 17 cells. AB - In the murine Ob 17 preadipocyte cell line, the thyroid hormone T3 is an adipogenic factor necessary at an early stage for differentiation into adipocyte. We demonstrate here that this T3 dependence may involve a transient expression (at both the messenger RNA and the protein levels) of c-ErbA beta-type receptors (T3R), although a large body of T3R remained the product of the c-erbAalpha gene, as previously described. c-ErbAbeta1 (and not beta2) expression emerged significantly at growth arrest, peaked 2 days later, and almost disappeared in maturing adipocytes. This expression is related to the presence of T3, as total deprivation of culture medium from T3 prevented it, and the addition of 1.5 nM T3 to preconfluent cultures was able to restore it. When cells were cultured in the presence of T3 and thus were able to differentiate, the c-erbAbeta peak was accompanied by sequential rapid increases in CAAT/enhancer-binding protein delta(C/EBPdelta), peroxisome proliferator-activated-gamma receptor (PPARgamma), and C/EBPalpha gene expressions. On the contrary, under thyroid hormone-deprived culture conditions that result in nondifferentiation of the preadipocytes, c erbAbeta1, PPARgamma, and the large C/EBPalpha expressions were blunted, and a moderate early increase in c-erbAalpha1 transcripts was sustained for a longer period. Addition of T3 to T3-deprived preconfluent cells restored PPARgamma and C/EBPalpha expressions. Taken together, the results highlight the important role of T3 in the adipogenesis of Ob 17 cells through the involvement of both beta1 and alpha1 T3R subtypes. PMID- 10385391 TI - Arachidonic acid directly mediates the rapid effects of 24,25-dihydroxyvitamin D3 via protein kinase C and indirectly through prostaglandin production in resting zone chondrocytes. AB - Prior studies have shown that 24,25-dihydroxyvitamin D3 [24,25-(OH)2D3] plays a major role in resting zone chondrocyte differentiation and that this vitamin D metabolite regulates both phospholipase A2 and protein kinase C (PKC) specific activities. Arachidonic acid is the product of phospholipase A2 action and has been shown in other systems to affect a variety of cellular functions, including PKC activity. The aim of the present study was to examine the interrelationship between arachidonic acid and 24,25-(OH)2D3 on markers of proliferation, differentiation, and matrix production in resting zone chondrocytes and to characterize the mechanisms by which arachidonic acid regulates PKC, which was shown previously to mediate the rapid effects of 24,25-(OH)2D3 and arachidonic acid on these cells. Confluent, fourth passage resting zone cells from rat costochondral cartilage were used to evaluate these mechanisms. The addition of arachidonic acid to resting zone cultures stimulated [3H]thymidine incorporation and inhibited the activity of alkaline phosphatase and PKC, but had no effect on proteoglycan sulfation. In contrast, 24,25-(OH)2D3 inhibited [3H]thymidine incorporation and stimulated alkaline phosphatase, proteoglycan sulfation, and PKC activity. In cultures treated with both agents, the effects of 24,25-(OH)2D3 were reversed by arachidonic acid. The PKC isoform affected by arachidonic acid was PKCalpha; cytosolic levels were decreased, but membrane levels were unaffected, indicating that translocation did not occur. Arachidonic acid had a direct effect on PKC in isolated plasma membranes and matrix vesicles, indicating a nongenomic mechanism. Plasma membrane PKCalpha was inhibited, and matrix vesicle PKCzeta was stimulated; these effects were blocked by 24,25-(OH)2D3. Studies using cyclooxygenase and lipoxygenase inhibitors indicate that the effects of arachidonic acid are due in part to PG production, but not to leukotriene production. This is supported by the fact that H8-dependent inhibition of protein kinase A, which mediates the effects of PGE2, had no effect on the direct action of arachidonic acid but did mediate the role of arachidonic acid in the cell response to 24,25-(OH)2D3. Diacylglycerol does not appear to be involved, indicating that phospholipase C and/or D do not play a role. Gamma linolenic acid, an unsaturated precursor of arachidonic acid, elicited a similar response in matrix vesicles but not plasma membranes, whereas palmitic acid, a saturated fatty acid, had no effect. These data suggest that arachidonic acid may act as a negative regulator of 24,25-(OH)2D3 action in resting zone chondrocytes. PMID- 10385392 TI - Thyroid hormone (3,5,3'-triido-L-thyronine) masking/inversion of stimulatory effect of androgen on expression of mk1, a true tissue kallikrein, in the mouse submandibular gland. AB - We studied hormonal regulation of the expression of mkl, a true tissue kallikrein, in the submandibular gland (SMG) of ICR, C3H/ HeN, and F1 (mice from male C3H/HeN x female ICR and in the ones from male ICR x female C3H/HeN). In these mouse strains, mk1 was low in content in males, abundant in females, and increased remarkably by castration of males. In the case of ICR and both F1 mice, injection of 5alpha-dihydrotestosterone (DHT) reduced the mkl level of castrated and female mice. However, the mkl content in female C3H/ HeN mice (or castrated C3H/HeN) was further increased by DHT. To investigate the real action of DHT on mk1 expression, we examined the effects of adrenoectomy/glucocorticoid (dexamethasone, Dex) administration; DHT administration into castrated and adrenoectomized mice; ovariectomy/female hormone (17beta-estradiol, progesterone) administration; and hypophysectomy/combinatory administration of DHT, Dex, and thyroid hormone (3,5,3'-triiodo-L-thyronine, T3) on the mk1 expression in the SMG of ICR mice. Adrenoectomy or ovariectomy did not change the characteristic pattern of mk1 expression in male and female ICR mice. In hypophysectomized (Hypox) ICR male mice, the mk1 content was increased to the same level as in normal ICR females, and DHT administration into the Hypox mice further increased the mk1 level. However, combinatory administration of DHT + T3 or of DHT + T3 + Dex into the Hypox mice lowered the mkl content to the level of normal ICR males, whereas T3 single administration had no effect. Dex single administration into the Hypox mice increased the mkl level to an even higher than that observed with DHT administration. The mk1 level in Hypox mice was not significantly changed by coadministration of Dex with T3. From these results, we conclude that 1) mk1 expression is fundamentally stimulated by androgen (DHT) as are other mk isozymes, such as mk9, mk13, mk22, and mk26 in the mouse SMG, 2) the effect (stimulatory) of DHT on mk1 expression becomes, however, inverted (inhibitory) in the presence of T3. Although the serum T3 level of C3H/HeN female (0.52 ng/ml) was not significantly different from that of C3H/HeN males or ICR mice, coadministration of T3 into C3H/HeN females with a fixed amount of DHT (20 mg/kg body weight) dose dependently repressed the DHT-induced increase in mkl expression, suggesting the lower sensitivity of C3H/HeN females to T3. PMID- 10385393 TI - Regulation and expression of gonadotropin-releasing hormone gene differs in brain and gonads in rainbow trout. AB - The GnRH gene is transcribed in both the brain and gonads. GnRH in the brain is critical for reproduction, but the function and importance of GnRH in the ovary and testis is not clear. In this study we examine whether regulation of the GnRH gene is distinct in the brain and gonads, whether the regulation of the GnRH gene in the gonads is altered after genome duplication, and whether the regulatory region of the GnRH gene is tightly conserved in vertebrates. From ovary and testis, we isolated and sequenced for the first time two different genes and their complementary DNAs that encode the identical peptide known as salmon GnRH. Rainbow trout were selected because they are tetraploid due to genome duplication. A downstream promoter is used in the brain and gonads by salmon GnRH messenger RNA1 (mRNA1) and mRNA2, but mRNA2 also uses an upstream promoter only in the gonads. Two types of long mRNA2 transcripts in ovary and testis both use an alternative start site at position 323; one of these types also retains intron 1. This long 5'-untranslated region is a likely site for distinct regulation of mRNA in the gonad. Additional evidence for separate regulation is that a different expression pattern exists in brain and gonads for GnRH mRNAs during development and maturation. Gene duplication did not alter the encoded peptide, but changed the expression pattern and resulted in complete divergence of the promoter sequence from position -215. A comparison of the mammalian and trout GnRH genes reveals that the promoters are without sequence identity except for a few consensus sites in both regulatory regions. The duplicated trout genes provide a model to study a critical gene whose product controls reproduction in all vertebrates. PMID- 10385394 TI - Alpha-latrotoxin stimulates inward current, rise in cytosolic calcium concentration, and exocytosis in at pituitary gonadotropes. AB - Alpha-latrotoxin (LTX) from the black widow spider venom, stimulates neurotransmitter release from neuronal cells via Ca2+ -dependent as well as Ca2+ independent mechanisms. In some peptide-secreting endocrine cells, however, LTX stimulates hormone release mainly via a Ca2+ -independent mechanism. Here we investigated the action of LTX in rat pituitary gonadotropes that secrete the peptide, LH. Using the patch-clamp technique in conjunction with the fluorescent Ca2+ indicator (indo-1) to simultaneously measure the cytosolic Ca2+ concentration ([Ca2+]i) and ionic current, we showed that LTX elicited bursts of inward current that were accompanied by [Ca2+]i elevations. In the presence of a physiological concentration of extracellular Ca2+, the unitary conductance of the LTX-induced current was about 300 pS, and only about 6.4% of the current was carried by Ca2+. The LTX-induced current was occasionally followed by intracellular Ca2+ release. At [Ca2+]i of 1 microM or more, exocytosis (detected by membrane capacitance measurement) was consistently triggered, and it was frequently followed by endocytosis. Thus, LTX triggers Ca2+ -dependent exocytosis in gonadotropes via extracellular Ca2+ entry as well as intracellular Ca2+ release. In approximately 25% of the cells, LTX could also trigger a slow exocytosis in the absence of [Ca2+]i elevation. Therefore, LTX has both Ca2+ dependent and Ca2+ -independent actions in gonadotropes. PMID- 10385395 TI - Steroidogenic factor-1 mediates cyclic 3',5'-adenosine monophosphate regulation of the high density lipoprotein receptor. AB - The high density lipoprotein (HDL) receptor mediates the uptake of cholesterol and cholesteryl esters, substrates for steroidogenesis, from an HDL particle in the adrenal gland and gonads. We report here that treatment of rat luteal cells with 1 mM (Bu)2cAMP for 24 h dramatically induced (118-fold) HDL receptor messenger RNA levels. The rat HDL receptor promoter contains a steroidogenic factor-1 (SF-1)-binding site (SFBd; 5'-TCAAGGCC-3') through which SF-1 protein binds and activates transcription of this gene in both human HTB9 bladder carcinoma and mouse Y1 tumor cells, an effect that is enhanced by cAMP. These observations demonstrate that this motif is required for both basal and cAMP induced regulation of the HDL receptor gene. Cotransfection studies in Kin 8 cells, a Y1 cell line resistant to cAMP activation as a result of a mutation in the protein kinase A (PKA) regulatory subunit, showed that a functional PKA is required for cAMP induction of HDL receptor gene transcription. Deleting the activation function-2 domain (amino acids 448-461) or mutating Ser430, a potential consensus phosphorylation site for PKA in the SF-1 protein, decreased both basal and cAMP-induced activation of the HDL receptor promoter. These data suggest that these regions within the SF-1 protein are required for both basal and cAMP-induced regulation of the HDL receptor gene. The mediation of cAMP responsiveness of the HDL receptor gene by SF-1 suggests how important this trans acting factor is in steroid hormone synthesis by assuring that all required elements (substrate and enzymes) are present when they are needed for maximal steroid production. PMID- 10385396 TI - Dysfunction of the islet lysosomal system conveys impairment of glucose-induced insulin release in the diabetic GK rat. AB - Accumulated evidence links an important signal involved in glucose-stimulated insulin release to the activation of the islet lysosomal glycogenolytic enzyme acid glucan-1,4-alpha-glucosidase. We have analyzed the function of the lysosomal system/lysosomal enzyme activities in pancreatic islets of young (6-8 weeks), spontaneously diabetic, GK (Goto-Kakizaki) rats and Wistar control rats in relation to glucose-induced insulin release. The insulin secretory response to glucose was markedly impaired in the GK rat, but was restored by the adenylate cyclase activator forskolin. Islet activities of classical lysosomal enzymes, e.g.. acid phosphatase, N-acetyl-beta-D-glucosaminidase, beta-glucuronidase, and cathepsin D, were reduced by 20-35% in the GK rat compared with those in Wistar controls. In contrast, the activities of the lysosomal alpha-glucosidehydrolases, i.e.. acid glucan-1,4-alpha-glucosidase and acid alpha-glucosidase, were increased by 40-50%. Neutral alpha-glucosidase (endoplasmic reticulum) was unaffected. Comparative analysis of liver tissue showed that lysosomal enzyme activities were of the same magnitude in GK and Wistar rats. Notably, in Wistar rats, the activities of acid glucan-1,4-alpha-glucosidase and acid alpha glucosidase were approximately 15-fold higher in islets than in liver. Other lysosomal enzymes did not display such a difference. Normalization of glycemia in GK rats by phlorizin administered for 9 days did not influence either the lysosomal alpha-glucosidehydrolase activities or other lysosomal enzyme activities in GK islets. Finally, the pseudotetrasaccharide acarbose, which accumulates in the lysosomal system, inhibited acid glucan-1,4-alpha-glucosidase activity in parallel with its inhibitory action on glucose-induced insulin release in intact Wistar islets, whereas no effect was recorded for either parameter in intact GK islets. In contrast, acarbose inhibited the enzyme activity equally in islet homogenates from both GK and Wistar rats, showing that the catalytic activity of the enzyme itself in disrupted cells was unaffected. We propose that dysfunction of the islet lysosomal/vacuolar system is an important defect impairing the transduction mechanisms for glucose-induced insulin release in the GK rat. PMID- 10385397 TI - Tumor necrosis factor-alpha stimulates lactate dehydrogenase A expression in porcine cultured sertoli cells: mechanisms of action. AB - In the present study, we investigated the regulatory action of tumor necrosis factor-alpha (TNFalpha) on lactate dehydrogenase A (LDH A), a key enzyme involved in lactate production. To this end, use was made of a primary culture system of porcine testicular Sertoli cells. TNFalpha stimulated LDH A messenger RNA (mRNA) expression in a dose (ED50 = 2.5 ng/ml; 0.1 nM TNFalpha)-dependent manner. This stimulatory effect was time dependent, with an effect detected after 6 h of TNFalpha treatment and maximal after 48 h of exposition (5-fold; P<0.001). The direct effect of TNFalpha on LDH A mRNA could not be accounted for by an increase in mRNA stability (half-life = 9 h), but was probably due to an increase in LDH A gene transcription. Inhibitors of protein synthesis (cycloheximide), gene transcription (actinomycin D and dichlorobenzimidazole riboside), tyrosine kinase (genistein), and protein kinase C (bisindolylmaleimide) abrogated completely (actinomycin D, dichlorobenzimidazole riboside, cycloheximide, and genistein) or partially (bisindolylmaleimide) TNFalpha-induced LDH A mRNA expression. These observations suggest that the stimulatory effect of TNFalpha on LDH A mRNA expression requires protein synthesis and may involve a protein tyrosine kinase and protein kinase C. In addition, we report that LDH A mRNA levels were increased in Sertoli cells treated with FSH. However, although the cytokine enhances LDH A mRNA levels through increased gene transcription, the hormone exerts its stimulatory action through an increase in LDH A mRNA stability. The regulatory actions of the cytokine and the hormone on LDH A mRNA levels and therefore on lactate production may operate in the context of the metabolic cooperation between Sertoli and postmeiotic germ cells in the seminiferous tubules. PMID- 10385398 TI - Insulin-like growth factor I protects oligodendrocytes from tumor necrosis factor alpha-induced injury. AB - Tumor necrosis factor-alpha (TNF-alpha) has been causally implicated in several demyelinating disorders, including multiple sclerosis. Because insulin-like growth factor I (IGF-I) is a potent stimulator of myelination, we investigated whether it can protect oligodendrocytes and myelination from TNF-alpha-induced damage using mouse glial cultures as a model. Compared with controls, TNF-alpha decreased oligodendrocyte number by approximately 40% and doubled the number of apoptotic oligodendrocytes and their precursors. Addition of Boc-aspartyl(Ome) fluoromethyl ketone (BAF), an inhibitor of interleukin-1beta converting enzyme (ICE)/caspase proteases, blocked TNF-alpha-induced reductions in oligodendrocytes, indicating that the TNF-alpha-induced reduction in oligodendrocytes is, at least in part, due to apoptosis, and that ICE/caspases are one of TNF-alpha action mediators. Simultaneous addition of IGF-I to TNF alpha-treated cultures negated these TNF-alpha effects nearly completely. Furthermore, IGF-I promoted oligodendrocyte precursor proliferation and/or differentiation in TNF-alpha-treated cultures. To analyze TNF-alpha and IGF-I actions on oligodendrocyte function, we measured the abundance of messenger RNAs (mRNAs) for two major myelin-specific proteins, myelin basic protein (MBP) and proteolipid protein (PLP). While TNF-alpha decreased MBP and PLP mRNA abundance by 5- to 6-fold, IGF-I abrogated TNF-alpha-induced reductions in a dose- and time dependent manner. The changes in MBP and PLP mRNA abundance could not be completely explained by the changes in oligodendrocyte number, indicating that myelin protein gene expression is regulated by both TNF-alpha and IGF-I. These data support the hypothesis that TNF-alpha can mediate oligodendrocyte and myelin damage, and indicate that IGF-I protects oligodendrocytes from TNF-alpha insults by blocking TNF-alpha-induced apoptosis, and by promoting oligodendrocyte and precursor proliferation/differentiation and myelin protein gene expression. PMID- 10385399 TI - Growth hormone-mediated regulation of insulin-like growth factor I promoter activity in C6 glioma cells. AB - The molecular mechanisms by which GH regulates insulin-like growth factor (IGF-I) gene expression remain obscure. One difficulty has been the lack of established GH-responsive cell lines that express the IGF-I gene. To develop such a cell line, we used rat C6 glioma cells which, as determined by RNase protection assay, express the IGF-I gene but not the GH receptor gene. To confer GH responsiveness, C6 cells were cotransfected with vectors that express the GH receptor (pRc/CMV WTrGHR) and Jak2 (pRc/CMV Jak2). GH responsiveness was demonstrated using luciferase reporter genes containing either the Sis-inducible element from the c fos gene (pTK81-SIE-Luc) or 6 copies of the GH-responsive GAS-like element (GLE) from the rat spi2.1 gene (pSpi-GLE-Luc). The SIE is activated by binding of STAT1 and 3, whereas the GLE binds STAT5. In cells cotransfected with pRc/CMV WTrGHR, pRc/CMV Jak2, and either pTK81-SIE-Luc or pSpi GLE-Luc, treatment with 500 ng/ml GH for 24 h stimulated a 3.1- and 1.7-fold increase in luciferase activity, respectively. These data suggest that in C6 cells cotransfected with pRc/CMV WTrGHR and pRc/CMV Jak2, GH activates STAT1, 3, and 5. To determine whether GH responsive IGF-I promoter activity could be demonstrated, C6 cells were cotransfected with pRc/CMV WTrGHR, pRc/ CMV Jak2, and an IGF-I-luciferase fusion gene that contained a fragment of the rat IGF-I gene that extended from -412 in the 5'-flanking region of exon 1 to the Met-22 in exon 3. GH stimulated a modest, but reproducible, 1.7-fold increase in luciferase activity in these cells, suggesting that a GH-responsive element is present in this region of the IGF-I gene. To better localize the GH-responsive element, cells were cotransfected with pRc/CMV WTrGHR, pRc/CMV Jak2 plus one of several IGF-I-luciferase fusion genes containing either fragments of one of the two promoters in the IGF-I gene or a fragment of intron 2 that includes a GH-responsive DNase I hypersensitivity site. For all constructs, treatment with GH for 24 h did not stimulate a significant increase in luciferase activity, suggesting that GH-responsive sequences are not located in these specific regions of the IGF-I gene or that GH-directed transcription of the IGF-I gene is mediated via several different regions of the IGF-I gene and the effect of any one of these regions in isolation was not sufficiently robust to be detected in this model system. In summary, transient expression of the GH receptor and Jak2 in C6 cells creates a GH-responsive system that activates STAT1, 3, and 5. Moreover, a fragment of the IGF-I gene that contains exons 1 and 2, a fragment of exon 3, and introns 1 and 2 is GH responsive using this model system. PMID- 10385400 TI - Basic fibroblast growth factor induces proteolysis of secreted and cell membrane associated insulin-like growth factor binding protein-2 in human neuroblastoma cells. AB - Insulin-like growth factor (IGF) action in the brain is modulated by IGF-binding proteins (IGFBPs) whose abundance can be altered by other locally expressed growth factors. However, the mechanisms involved are unclear. We here employed the neuroblastoma cell line SK-N-MC as a model to define the mechanisms involved in modulation of IGFBPs in neuronal cells. Western ligand blotting analysis and immunoprecipitation of conditioned media (CM) from SK-N-MC cells showed that in these cells, as in the brain, the most abundantly expressed IGFBP was IGFBP-2. However, IGFBP-2 was barely detectable in CM from cells treated with basic fibroblast growth factor (bFGF) without a change in IGFBP-2 messenger RNA (mRNA) abundance. These CM contained specific IGFBP-2 proteolytic activity, resulting in two IGFBP-2 fragments of 14 and 22 kDa. The activity was inhibited by EDTA/phenylmethylsulfonyl fluoride or aprotinin. Competitive binding studies indicated that IGFBP-2 fragments had reduced binding affinity for IGF-I. bFGF induced IGFBP-3 mRNA and protein. Affinity cross-linking of [125I]IGF-I to neuroblastoma cell membranes followed by immunoprecipitation revealed a approximately 38 kDa [125I]IGF-I/IGFBP-2 complex. Cell surface-associated IGFBP-2 was also susceptible to bFGF-induced proteolysis, with the appearance of a single cross-linked 21-kDa complex with low affinity for IGF-I. These findings indicate that intact IGFBP-2 and the 14-kDa, but not the 22-kDa fragment, bind to the cell surface. Our data suggest that induction of IGFBP-2 proteolysis on neuronal cell surface is a novel mechanism whereby IGF availability is modulated by the local growth factor bFGF. PMID- 10385401 TI - Effect of retinoic acid on glucokinase activity and gene expression and on insulin secretion in primary cultures of pancreatic islets. AB - Retinoic acid has manifold effects on pancreatic beta-cells. Previously we reported that retinoic acid increases glucokinase activity and messenger RNA (mRNA) levels in the insulinoma cell line RIN-m5F; however, we could not rule out the possibility that the effect of retinoic acid on RIN-m5F glucokinase was inherent to the cell line or related to its differentiating capacity. In this report, we demonstrate that physiologic concentrations of retinoic acid stimulate glucokinase activity in both fetal islets and differentiated adult islets in culture. In the adult tissue, the response to the retinoid was less pronounced, achieving about half of the maximal effect produced on the fetal tissue. Using the branched DNA (bDNA) assay, a sensitive signal amplification technique, we detected relative increases in glucokinase mRNA levels of 51.8+/-13.3% and 62.8+/ 16.1% at 12 and 24 h, respectively, in adult islets treated with] 10(-6) M retinoic acid. In fetal islets, increases of 55+/-14.9% and 107+/-30.5% at 12 and 24 h, respectively, were observed. In transfected fetal islets, retinoic acid increased the activity of the -1000 kb rat glucokinase promoter by 51.3%. Because glucokinase activity controls insulin secretion, we also investigated the effect of retinoic acid on insulin secretion. Treatment with 10(-6) M retinoic acid for 24 h increased insulin secretion in both fetal and adult islets; however, the increases on insulin secretion were more pronounced in the mature islets; in contrast, retinoic acid produced higher levels of insulin mRNA in the fetal islets. These data show that retinoic acid increases pancreatic glucokinase in cultured islets and that the mechanism may involve a stimulatory effect on the glucokinase promoter. PMID- 10385402 TI - Transcription activating and repressing functions of the androgen receptor are differentially influenced by mutations in the deoxyribonucleic acid-binding domain. AB - Despite the wide spectrum of androgen receptor (AR) mutants described in androgen insensitivity syndromes (AIS), their influence on transactivating and, in particular, transrepressing functions of AR are poorly defined. Rat AR mutants with substitutions in the DNA-binding domain, corresponding to several mutations in AIS patients, were examined for these activities. AR variants (G551V and C562G) with mutations in the first zinc finger (ZF) exhibited reduced DNA binding activity and attenuated transactivation. An R590Q substitution in the second ZF diminished transcriptional activity only from a promoter with a single androgen response element, whereas activation at multiple androgen response element sites was unaffected, despite the poor DNA-binding affinity of R590Q. Another substitution in the second ZF, A579T, yielded similar findings. In comparison to wild-type AR, G551V, and C562G variants had markedly reduced ability to repress an NF-kappaB/RelA-activated promoter but R590Q behaved like the native receptor. AP1 function was repressed not only by wild-type AR but also by the transactivating mutants A579T and R590Q as well as by the transcriptionally inactive mutants G551V and C562G. Furthermore, a Lys-to-Ala substitution in codon 563 of the first ZF switched AR into a ligand-dependent activator at AP1 sites but maintained the ability to repress NF-kappaB/RelA function. Taken together, DNA-binding domain mutations in AIS patients influence transcriptional activating and repressing functions of AR in a selective fashion, which probably contributes to the complexity in the presentation of the AIS phenotype. PMID- 10385403 TI - Hepatocyte growth factor and c-MET are expressed in rat prepuberal testis. AB - The hepatocyte growth factor (HGF) receptor (c-MET) is present in different mammalian tissues and transduces multiple biological effects. The HGF is known to regulate many fundamental cellular functions, such as cell growth, movement and differentiation, and is involved in embryonal morphogenesis. We have studied HGF and c-MET expression in prepuberal rat testis. c-MET gene expression was found in total testis and in homogeneous cell populations, as demonstrated by Northern blotting. In the seminiferous tubules, c-MET gene was only expressed in the myoid cells. In these cells, c-MET was detectable and constantly expressed for at least six days of culture. The interstitial tissue was also c-MET positive. The protein encoded by the MET proto-oncogene was detected in myoid cells, and HGF administration to these cells induced morphological changes in the cells. HGF expression was not detected by Northern blotting using RNA extracted from total testis. By contrast, when homogenous cell populations were used, HGF expression was detectable and exclusively localized in myoid cells. Myoid cell-conditioned medium was able to induce scattering of canine kidney epithelial (MDCK) cells, and the scatter effect of a 3-days conditioned medium was evident even after 7 fold dilution of the medium. Our findings demonstrate that HGF and its receptor are present in rat prepuberal testis. The coexpression of factor and receptor in the myoid cells suggests a new role for HGF as autocrine regulator of myoid cell function and, possibly, as regulator of mammalian testicular function. PMID- 10385404 TI - Homologous androgen receptor up-regulation in osteoblastic cells may be associated with enhanced functional androgen responsiveness. AB - Although androgens have myriad effects on the skeleton, the regulation of androgen action in bone is not well understood. Androgen receptors (ARs) are known to play an important role in mediating androgen action. We have examined the effects of androgens and other sex steroids on AR levels in osteoblastic cells in vitro using two clonal human cell lines, SaOS-2 and U-2 OS. AR protein levels were quantitated both by specific androgen binding studies and Western analyses, and AR messenger RNA was measured with RNase protection assays. Potential changes in AR functionality was assessed by reporter assays. Treatment of osteoblastic cells with the nonaromatizable androgen 5alpha dihydrotestosterone (DHT) increased specific androgen binding 2-to 4-fold. Similar increases in AR protein levels were documented by Western analysis in both cell lines. The androgen-mediated increase in receptor levels was time and dose dependent as well as androgen specific. Steady-state AR messenger RNA levels were also increased by DHT. When AR concentrations in osteoblastic cells were elevated with exogenous receptor, there was an enhancement of DHT responsiveness, measured by increased trans-activation of an androgen-responsive promoter. Thus, androgen exposure increased androgen receptor protein levels and specific androgen binding in osteoblastic cells. Androgen action as measured by androgen mediated transcriptional activation is enhanced in the presence of elevated AR levels. Consequently, these studies have revealed an additional means by which androgens may modulate skeletal metabolism. PMID- 10385405 TI - Interruption of activin A autocrine regulation by antisense oligodeoxynucleotides accelerates liver tumor cell proliferation. AB - Administration of activin A, a member of the transforming growth factor-beta superfamily inhibits hepatocyte proliferation in vitro and reduces liver mass in vivo. However, a role of endogenous activin A in local growth modulation has not been established in any system. The aim of this study was to examine the production of activin A in the human hepatoma cell line HLF and to explore a possible autocrine role of activin as a cell growth inhibitor by blocking production of endogenous activin using antisense oligodeoxynucleotides. Administration of exogenous activin A suppressed HLF cell growth, and immunoreactive activin A was shown to be produced in the cells at confluency by Western blotting analysis. Cells were exposed to phosphorothioate-modified oligodeoxynucleotides, synthesized with antisense or randomly shuffled base sequences of activin betaA subunit messenger RNA, under serum-free conditions. Uptake of the oligodeoxynucleotides into the cells was confirmed by use of fluorescein isothiocyanate-labeled oligodeoxynucleotides. Administration of antisense oligodeoxynucleotides reduced activin A production as confirmed by both competitive PCR and Western blotting. Activin betaA antisense oligodeoxynucleotides significantly increased cell proliferation compared with controls. These findings are consistent with the existence of an autocrine role of activin A as an inhibitor of hepatocyte proliferation. PMID- 10385406 TI - Short photoperiods evoke testicular apoptosis in white-footed mice (Peromyscus leucopus). AB - Many small, nontropical mammals stop breeding during winter. Chronic exposure of males to short days (<12.5 h light/day) causes the testes to atrophy and both steroidogenesis and gametogenesis to decrease. Male white-footed mice (Peromyscus leucopus) exposed to inhibitory short day lengths provide a natural animal model to study the cellular mechanisms regulating testicular regression. In the present study, the possible role of apoptosis was assessed during naturally occurring, short day-induced gonadal regression in white-footed mice by in situ terminal transferase-mediated end labeling (TUNEL), quantitative DNA 3'-end-labeling autoradiography (laddering) of DNA fragments, and quantification of Fas protein expression, an early initiator of apoptosis. Sexually mature male mice were exposed to short (8 h of light, 16 h of darkness) or long (16 h of light, 8 h of darkness) day lengths for 2, 4, 6, 8, or 10 weeks; gonads were then removed and processed for detection of apoptotic activity. In common with previous studies, the first significant reduction in relative testis mass was observed at week 10 of short day exposure. A 2- to 3-fold increase in apoptotic (TUNEL-positive) germ cells per seminiferous tubule was observed in the testes of mice exposed to short days for 4, 6, 8, or 10 weeks compared with the testes of long day animals. The extent of 3'-end labeling of low mol wt DNA increased with 4-8 weeks of short day exposure. Western blot analysis revealed an up-regulation of the Fas protein in the testes of short day males at 4, 8, and 10 weeks. Fas staining was primarily localized to spermatocytes and spermatids. Plasma testosterone concentrations decreased in short compared with long day animals after 6, 8, or 10 weeks. The increase in TUNEL positive-labeled germ cells, testicular DNA fragmentation, and up-regulation of the Fas protein before short day reductions of testis mass and function suggest that apoptosis is important for the mediation of photoperiod induced testicular regression in white-footed mice. PMID- 10385407 TI - FKHR binds the insulin response element in the insulin-like growth factor binding protein-1 promoter. AB - The insulin response element (IRE) in the IGFBP-1 promoter, and in other gene promoters, contains a T(A/G)TTT motif essential for insulin inhibition of transcription. Studies presented here test whether FKHR may be the transcription factor that confers insulin inhibition through this IRE motif. Immunoblots using antiserum to the synthetic peptide FKHR413-430, RNase protection, and Northerns blots show that FKHR is expressed in HEP G2 human hepatoma cells. Southwestern blots, electromobility shift assays, and DNase I protection assays show that Escherichia coli-expressed GST-FKHR binds specifically to IREs from the IGFBP-1, PEPCK and TAT genes; however, unlike HNF3beta, another protein proposed to be the insulin regulated factor, GST-FKHR does not bind the insulin unresponsive G/C-A/C mutation of the IGFBP-1 IRE. When HEP G2 cells were cotransfected with FKHR expression vectors and with IGFBP-1 promoter plasmids containing either native or mutant IREs, FKHR expression induced a 5-fold increase in activity of the native IGFBP-1 promoter but no increase in activity of promoter constructs containing insulin unresponsive IRE mutants. These data suggest that FKHR, and/or a related family member, is the important T(G/A)TTT binding protein that confers the inhibitory effect of insulin on gene transcription. PMID- 10385408 TI - The effect of hypoxia from birth on the regulation of aldosterone in the 7-day old rat: plasma hormones, steroidogenesis in vitro, and steroidogenic enzyme messenger ribonucleic acid. AB - Adaptation to hypoxia in the neonate requires an appropriate adrenocortical response. The purpose of this study was to examine the adaptation of the aldosterone pathway in rat pups exposed to hypoxia in vivo from birth to 7 days of age. Neonatal rats (with their lactating dams) were exposed to normoxia (21% O2) or hypoxia (12% O2) continuously for 7 days from birth. Trunk blood was collected, and entire adrenal glands were processed from 7-day-old rats to study the activity of the steroidogenic pathway in dispersed cells and isolated mitochondria, for measurement of expression of the steroidogenic enzyme messenger RNAs (mRNAs) by RT-competitive PCR and in situ hybridization histochemistry, for measurement of zona glomerulosa width by immunohistofluorescent staining for P450c11AS protein, and for measurement of mitochondrial number and distribution by transmission electron microscopy. Exposure to hypoxia for 7 days from birth resulted in a marked increase in plasma ACTH, corticosterone, and aldosterone with no change in PRA. Aldosteronogenesis and P450c11AS activity were both augmented in dispersed cells; this effect was lost in isolated mitochondria (from entire adrenal glands) using a permeable substrate for P450c11AS. There was no significant effect of hypoxia on expression of the steroidogenic enzyme mRNAs measured by RT-competitive PCR or in situ hybridization histochemistry. Finally, hypoxia had no effect on mitochondrial number or stereology as assessed by transmission electron microscopy or on zona glomerulosa width as assessed by staining for P450c11AS protein. We conclude that, as opposed to that in adults, hypoxia in the neonate results in an augmentation of aldosteronogenesis. This effect is not accounted for by a change in steroidogenic enzyme mRNA expression, zona glomerulosa width (i.e. hyperplasia), or mitochondrial number or distribution. This functional augmentation of aldosteronogenesis may be due to a change in mitochondrial permeability to steroid substrates and/or the effect of cytosolic factors that control mitochondrial steroidogenesis. PMID- 10385409 TI - 3',5'-Cyclic adenosine monophosphate activation in osteoblastic cells: effects on parathyroid hormone-1 receptors and osteoblastic differentiation in vitro. AB - PTH has anabolic and catabolic effects in bone through activation of the PTH-1 (PTH/PTHrP) receptor and the cAMP/protein kinase A pathway. The effects of agents that regulate cAMP in nontransformed osteoblasts in relation to cell differentiation have not been described. The purpose of this study was to determine the effects of PTH fragments with differing cAMP-stimulating activity, and nonPTH cAMP regulators on PTH-1 receptor expression and activity, and osteoblast differentiation in vitro using MC3T3-E1 and primary rat calvarial cells. PTH (1-34), but not PTH (53-84), (7-34), or PTHrP (107-139) treatment (24 h) resulted in down-regulation of steady-state messenger RNA for the PTH-1 receptor. Forskolin (a stimulator of cAMP accumulation) also down regulated the PTH-1 receptor, whereas 9-(tetrahydro-2-furyl) adenine (THFA) (an inhibitor of adenylyl cyclase) had no effect. Similarly, PTH (1-34) treatment for 48 h abolished PTHrP binding to cell surface receptors; however, neither the PTH analogs nor the cAMP regulating agents altered PTH binding or numbers of binding sites on osteoblastic cells. Basal levels of cAMP were reduced in cultured cells treated for 6 days with PTH (7-34) or THFA compared with controls. In contrast, PTH-stimulated cAMP levels were significantly increased in cultures treated with PTH (7-34) and THFA for 6 days during osteoblast differentiation and were decreased in cultures treated with PTH (1-34) and forskolin compared with controls. To evaluate effects of the cAMP pathway on osteoblast differentiation, cultures were treated continuously with PTH analogs and cAMP regulators during an 18-day differentiation regime, total RNA was isolated at multiple time points, and Northern blot analysis for osteocalcin (OCN) was performed. THFA and PTH (7 34)-treated cultures had increased OCN expression; whereas, PTH (1-34) and forskolin reduced OCN expression. Interestingly, PTH (7-34) and THFA-treated cultures had increased mineralized nodule formation, in contrast to PTH (1-34) and forskolin treatment, which reduced nodule formation. Similarly, calcium accumulation in cultures was significantly increased in the PTH (7-34) and THFA treated cultures and reduced in the PTH (1-34) and forskolin-treated cultures. These data demonstrate that agents that increase cAMP down regulate PTH-1 receptor messenger RNA and inhibit osteoblast differentiation in vitro. Agents that reduce or block adenylyl cyclase or cAMP activity do not alter PTH-1 receptor expression or binding, but have striking effects on promoting osteoblast differentiation. We conclude that many effects of PTH on osteoblasts may be mimicked or antagonized by agents that alter cAMP activity and bypass the PTH-1 receptor. PMID- 10385410 TI - Prolonged activation of ERK2 by epidermal growth factor and other growth factors requires a functional insulin-like growth factor 1 receptor. AB - We have investigated the activation of ERK2, a serine/threonine kinase necessary for transmission of mitogenic signals, in cells derived from mouse embryos homozygous for a null mutation of the insulin-like growth factor (IGF)-1R gene (R cells) and from wild-type littermates (W cells), respectively. Stimulation of quiescent W cells with IGF-1, epidermal growth factor (EGF), or with a combination growth factors induced both a maximal transient and a prolonged activation of ERK2, whereas platelet-derived growth factor or a combination of platelet-derived growth factor and EGF resulted only in transient activation of ERK2. In contrast, stimulation of R cells with IGF-1, EGF, or combinations of growth factors resulted in a transient and submaximal activation of ERK2. Reintroduction of a wild-type human IGF-1R or of a C-terminus IGF-1R mutant, but not of a juxtamembrane mutant IGF-1R, into R- cells was able to restore ERK2 activation to wild-type levels. Thus, prolonged ERK2 activation in mouse embryo fibroblasts stimulated with purified growth factors is largely dependent on a signal generated by the IGF-1R. PMID- 10385411 TI - Regulation of rat cardiac Kv1.5 gene expression by thyroid hormone is rapid and chamber specific. AB - Thyroid hormone affects the contractile and electrophysiological properties of the cardiac myocyte that result in part from changes in the expression of thyroid hormone-responsive cardiac genes, including those that regulate membrane ion currents. To determine the molecular mechanisms underlying this effect, expression of a voltage-gated K+ channel, Kv1.5, was measured in response to thyroid hormone. Using quantitative RT-PCR methodology, the content of Kv1.5 messenger RNA (mRNA) in left ventricles of euthyroid rats was 4.25+/-0.6x10(-20) mol/microg total RNA and was decreased by 70% in the hypothyroid rat ventricle to 1.27+/-0.80x10(-20) mol/microg RNA (P<0.01). Administration of T3 to hypothyroid animals restored ventricular Kv1.5 mRNA to control levels within 1 h of treatment, making this the most rapid T3-responsive cardiac gene reported to date. The half-life of Kv1.5 mRNA was 1.9 h and 2.0 h in euthyroid and hypothyroid ventricles, respectively, and T3 treatment of the rats did not alter its half-life. In atrial myocardium, expression of Kv1.5 mRNA (6.10+/-0.37x10( 20) mol/microg RNA) was unaltered by thyroid hormone status. The myocyte-specific and chamber-selective expression of Kv1.5 mRNA was confirmed in primary cultures of rat atrial and ventricular myocytes. PMID- 10385412 TI - Regulation of alkaline phosphatase activity by p38 MAP kinase in response to activation of Gi protein-coupled receptors by epinephrine in osteoblast-like cells. AB - The signaling mechanisms responsible for the regulation of alkaline phosphatase (ALP) activity by exogenous factors in osteoblast-like cells remain poorly understood. Among various agents, epinephrine was recently found to increase ALP activity in differentiating MC3T3-E1 cells by stimulating alpha1 adrenergic receptors coupled to Gi proteins. In the present study, we investigated the role of both ERK2 and p38 mitogen-activated protein (MAP) kinases in mediating this response in MC3T3-E1 cells. Our results indicate that both MAP kinases are transiently stimulated by epinephrine in differentiating cells via a pertussis toxin sensitive mechanism. The role of each MAP kinase pathway in mediating the stimulation of ALP activity by epinephrine was investigated using specific inhibitors. The MEK inhibitor PD98059, blocked ERK2 activity induced by epinephrine but had no effect on the stimulation of ALP activity. In contrast, low concentrations of SB203580, a specific inhibitor of the p38 MAP kinase, completely blunted this cellular response. However, this inhibitor had no influence on the stimulation of ALP activity induced by ascorbic acid. In conclusion, the results of this study suggest distinct roles for ERK and p38 MAP kinase pathways in regulating activity of MC3T3-E1 osteoblastic cells. The ERK pathway is likely involved in the control of cell proliferation whereas the p38 MAP kinase pathway regulates ALP activity in response to activation of Gi protein coupled receptors. PMID- 10385413 TI - Selective dependence of intracerebroventricular neuropeptide Y-elicited effects on central glucocorticoids. AB - It has been reported that hyperphagia and excessive body weight gain of genetically obese rodents were abolished by adrenalectomy. High hypothalamic levels of neuropeptide Y (NPY) were found in obese rodents. A chronic intracerebroventricular (icv) infusion of NPY in normal rats was shown to produce most hormono-metabolic abnormalities of genetically obese animals, and to be inefficient in doing so in adrenalectomized (ADX) rats. The combined presence of NPY and of glucocorticoids thus appeared to be necessary for inducing obesity. This study, therefore, was aimed at determining the consequences of a chronic i.c.v. NPY infusion in ADX rats receiving or not i.c.v. glucocorticoids. It was found that the combined i.c.v. infusion of NPY and dexamethasone in ADX rats increased food intake, body weight, plasma insulin, leptin, and triglyceride levels relative to vehicle-infused ADX controls. The infusion of NPY alone, or of dexamethasone alone in ADX rats failed to produce these effects. In contrast, the icv infusion of NPY alone greatly decreased the expression of brown adipose tissue uncoupling protein-1 and -3. This was not modified by the superimposed infusion of dexamethasone. It is concluded that, although many of centrally elicited NPY effects require the central presence of glucocorticoids, those bearing on the inhibition of uncoupling proteins expression (energy dissipation) do not require central glucocorticoids. PMID- 10385414 TI - Differentiation of adipose stromal cells: the roles of glucocorticoids and 11beta hydroxysteroid dehydrogenase. AB - Glucocorticoids play an important role in determining adipose tissue distribution and function, with glucocorticoid excess states such as Cushing's syndrome resulting in central obesity. We have investigated the functional significance of local generation of cortisol within adipose tissue from inactive cortisone through the activity of the NADP(H)-dependent enzyme, 11beta-hydroxysteroid dehydrogenase type 1 (11betaHSD1). In primary cultures of paired omental (om) and sc human adipose stromal cells (ASC; n = 34), 11betaHSD1 oxo-reductase activity was significantly higher in om ASC (median, 40.2 pmol/mg protein x h; 95% confidence interval, 1.8-105) compared with sc ASC (median, 11.4; 95% confidence interval, 0-48.1; P<0.001) despite similar endogenous NADPH/NADP concentrations. Both cortisol and insulin increased the differentiation of ASC to adipocytes (as assessed by glycerol-3-phosphate dehydrogenase expression), but only cortisol increased 11betaHSD1 activity and messenger RNA levels in a dose-dependent fashion. Cortisone (500 nM) was as effective as 500 nM cortisol in inducing ASC differentiation, but this stimulatory effect was inhibited by coincubation with the 11betaHSD1 inhibitor, glycyrrhetinic acid. The higher local conversion of cortisone to active cortisol through expression of 11betaHSD1 in om compared with sc ASC may explain the specific action of glucocorticoids on different adipose tissue depots. 11betaHSD1 expression in om ASC is regulated at a transcriptional level and is increased by glucocorticoids, but is not entirely dependent upon ASC differentiation. Inhibition of 11betaHSD1 within om ASC inhibits cortisone induced ASC differentiation. These findings indicate that local metabolism of glucocorticoid may control differentiation of adipose tissue in a site-specific fashion. Specific inhibitors of 11betaHSD1 may offer a novel approach for the treatment of patients with central obesity. PMID- 10385415 TI - Effect of diazoxide on brain capillary insulin receptor binding and food intake in hyperphagic obese Zucker rats. AB - Insulin is believed to act as a central adiposity signal by binding to hypothalamic and other brain insulin receptors. Entry of circulating insulin into the brain is accomplished by a saturable receptor-mediated transendothelial transport system and is believed to be impaired in hyperinsulinemic, insulin resistant, and hyperphagic obese Zucker rats. Theoretically, if hyperinsulinemia is decreased simultaneously while brain capillary insulin binding is increased, uptake of insulin into the brain can be enhanced leading to reduced food intake. To test this hypothesis, we administered diazoxide (DZ, 150 mg/kg/day), an inhibitor of glucose-mediated insulin secretion, or vehicle (control) to 7-week old female obese and lean Zucker rats for 4 weeks (n = 24-28/subgroup-strain). Animals were assigned to either fasted (FD) or free-fed (FF) protocol for determination of plasma and cerebrospinal fluid (CSF) insulin and brain capillary insulin binding at the end of 4 weeks. DZ obese consumed fewer calories (P<0.01) and gained less weight than control obese (P<0.01), whereas DZ lean had similar amounts of caloric intake and weight gain compared with lean controls. DZ obese had lower fasting and random plasma glucose than control obese (P<0.05). FD and FF DZ-treated obese and lean rats had lower plasma insulin than their respective obese (P<0.01) and lean (P<0.01) controls. FD and FF DZ-treated obese rats demonstrated higher CSF insulin (P<0.05) and CSF/ plasma insulin ratio (P<0.01) than their controls, while only FF DZ lean animals showed higher CSF/plasma insulin ratio (P<0.01) than their controls (P<0.05). This was associated with enhanced brain capillary insulin binding in FD and FF DZ-treated obese (P<0.01) and lean (P<0.05) animals compared with their respective controls. It was concluded that DZ treatment in obese Zucker rats caused a decrease in insulin secretion and partially reversed impaired insulin binding to brain capillaries, leading to enhanced brain insulin uptake, and resulted in reduced food intake and weight gain observed in these animals. PMID- 10385416 TI - Growth hormone (GH) and prolactin receptors in human peripheral blood mononuclear cells: relation with age and GH-binding protein. AB - GH receptors (GHRs) and PRL receptors (PRLRs) were studied in human peripheral blood mononuclear cells (PBMC) using flow cytometry, biotinylated anti-GH receptor monoclonal antibody 10B8, and biotinylated human PRL. Variations of GHR and PRLR expression and the relationship of plasma GHBP and GH receptor in PBMC subsets were examined as a function of age and sex. By double immunofluorescence staining, we show that about 30% of total cells express GH receptors, with a low expression in T cells, whereas almost all B cells and monocytes are GH receptor positive. Four age groups were defined among the 64 normal volunteers, aged 12 to 85 yr, who were included in the study. The percentage of PBMC expressing GH receptors is significantly lower in group 2 (20-40 yr) than in group 1 (12-20 yr) and group 4 (>60 yr). In T cells, monocytes and B cells, no significant changes are detected in either the percentage of GH receptor positive cells or in the GH receptor level per cell. The level of PRLRs expressed in PBMC is significantly higher in age group 2 than in age group 4. A negative correlation is observed between plasma GHBP and the percentage of PBMC expressing GH receptors. These results suggest that regulation of GH receptors in lymphocytes and in other target cells could be different. PMID- 10385417 TI - Progesterone- and dexamethasone-dependent osteoprogenitors in bone cell populations derived from rat vertebrae are different and distinct. AB - Previous experiments have demonstrated that bone cell populations derived from explants of lumbar vertebral bone of adult female rats contain osteoprogenitors that require dexamethasone (Dex) or progesterone (Prog) to proliferate and differentiate into fully differentiated bone-forming osteoblasts. We now show that the Prog-dependent population cannot be detected in male rats after sexual maturation, but is present in prepubertal rats of both sexes and can be induced in adult male-derived populations by culturing the explants in medium containing 17beta-estradiol (10(-9)-10(-8) M). This suggested that the Prog- and Dex dependent osteoprogenitors in adult female-derived populations were probably distinct populations and that the survival of the Prog-dependent osteoprogenitors and/or their ability to proliferate are dependent on the presence of estrogen. We then proceeded to prove this by using replica plating. When one of the paired colonies duplicated was cultured in medium containing Dex (10(-8) M) and the other in medium containing Prog (10(-5) M), 5.0% of duplicates formed bone in Prog only, 11.1% formed bone in Dex only, and 3.4% formed bone in both Prog and Dex. In all cases the size of the bone-forming colonies in Dex-treated cultures was larger than that in Prog-treated cultures, indicating that the effects of Dex on osteoprogenitor proliferation are greater than those of Prog. The results demonstrate the existence of three classes ofosteoprogenitors in adult female rat derived bone cell populations: a class responding to Dex only, a class responding to Prog only, and a class responding to both Dex and Prog. The results also indicate that the effects of Prog are not mediated by Prog binding to the glucocorticoid receptor and imply that Prog plays an important role in maintaining bone mass through regulating the class of osteoprogenitors responsive to Prog. PMID- 10385418 TI - Tumor necrosis factor-alpha-activated cell death pathways in NIT-1 insulinoma cells and primary pancreatic beta cells. AB - Tumor necrosis factor-alpha (TNFalpha) is a potential mediator of beta cell destruction in insulin-dependent diabetes mellitus. We have studied TNF responsive pathways leading to apoptosis in beta cells. Primary beta cells express low levels of the type I TNF receptor (TNFR1) but do not express the type 2 receptor (TNFR2). Evidence for TNFR1 expression on beta cells came from flow cytometry using monoclonal antibodies specific for TNFR1 and TNFR2 and from RT PCR of beta cell RNA. NIT-1 insulinoma cells similarly expressed TNFR1 (at higher levels than primary beta cells) as detected by flow cytometry and radio-binding studies. TNF induced NF-kappaB activation in both primary islet cells and NIT-1 cells. Apoptosis in response to TNFalpha was observed in NIT-1 cells whereas apoptosis of primary beta cells required both TNFalpha and interferon-gamma (IFNgamma). Apoptosis could be prevented in NIT-1 cells by expression of dominant negative Fas-associating protein with death domain (dnFADD). Apoptosis in NIT-1 cells was increased by coincubation with IFNgamma, which also increased caspase 1 expression. These data show that TNF-activated pathways capable of inducing apoptotic cell death are present in beta cells. Caspase activation is the dominant pathway of TNF-induced cell death in NIT-1 cells and may be an important mechanism of beta cell damage in insulin-dependent diabetes mellitus. PMID- 10385419 TI - Selective up-regulation of phosphodiesterase-4 cyclic adenosine 3',5' monophosphate (cAMP)-specific phosphodiesterase variants by elevated cAMP content in human myometrial cells in culture. AB - In human myometrium, the modulation of intracellular cAMP content resulting from agonist-mediated stimulation of the receptor-adenylyl cyclase complex is largely influenced by the rate of cAMP hydrolysis by phosphodiesterase (PDE) isoenzymes. We have previously shown that the PDE4 family contributes to the predominant cAMP hydrolyzing activity in human myometrium and that elevation of the PDE4B2 messenger RNA steady state level occurs in pregnant myometrial tissue. In the present study, we used a model of human myometrial cells in culture to determine whether an elevated cAMP concentration could influence PDE expression. As in myometrial tissue, high levels of PDE4 activity were detected in these smooth muscle cells. Long term treatment with 8-bromo-cAMP or forskolin resulted in a selective induction of PDE4B and of PDE4D short form messenger RNA variants. Concurrently, an increased immunoreactive signal for the PDE4B- and PDE4D-related isoenzymes was detected. This induction was consistent with an observed significant up-regulation of PDE4 activity. Accordingly, our results demonstrate that in human cultured myometrial cells, cAMP-elevating agents manipulate PDE4 activity through selective induction of synthesis of PDE4B and PDE4D short forms. Such a mechanism might have physiological importance during pregnancy by dampening hormonal stimulation and could thereby be involved in tolerance to the tocolytic effect of beta-adrenoceptor agonists. PMID- 10385420 TI - c-fos activity in Rana esculenta testis: seasonal and estradiol-induced changes. AB - Estradiol-17beta (E2) is suspected to exert a role in the regulation of testicular activity. Using a nonmammalian vertebrate model (the frog, Rana esculenta), we have investigated whether c-fos activity is detectable in the testis during the annual sexual cycle and whether E2 exerts a regulatory role on spermatogenesis through fos activity. FOS protein is available in testicular nuclear extracts (about 60 kDa) and, surprisingly, also in cytosolic extracts (about 60, 80, and 100 kDa). Estradiol induces primary spermatogonia (ISPG) proliferation [this effect is counteracted by antiestrogens (Tamoxifen and ICI 182-780)] and FOS appearance in testicular cytosolic extracts as well as c-fos transcription. Also, this effect is counteracted by ICI 182-780. Interestingly, the number of FOS immunopositive nuclei of ISPG strongly increases after E2 treatment, whereas a great increase of immunopositivity in the cytoplasm of ISPG is observed with the contemporaneous treatment with antiestrogens. In conclusion, our results demonstrate that E2 induces ISPG multiplication in the frog, R. esculenta, and, for the first time in a vertebrate species, that it triggers c fos activity in the testis. Moreover, E2 may be involved in mechanisms related to FOS transport in the nucleus of ISPG to induce the mitotic activity. PMID- 10385421 TI - Thiazolidinedione effects on glucocorticoid receptor-mediated gene transcription and differentiation in osteoblastic cells. AB - The glucocorticoid receptor (GR) and peroxisome proliferator-activated receptors (PPARs) play important roles in the differentiation of mesenchymal cells. Glucocorticoids acting via the GR promote osteoblastic differentiation of bone marrow stromal cells, whereas PPAR ligands induce these cells to become adipocytes. To explore potential interactions between PPAR and GR pathways in osteoblasts, we studied the interaction between PPAR subtype-selective ligands and dexamethasone (DEX) in a murine calvaria-derived osteoblastic cell line (MB 1.8) that expresses endogenous GR and PPARs. In ligand-dependent transcription assays, the PPARgamma-selective ligand TZD [(5-(4-N-methyl-N(2 pyridyl)amino)ethoxy)benzyl)thiazolidine-2,4-dione], a thiazolidinedione antidiabetic, enhanced the effect of DEX to stimulate transcription of a glucocorticoid-inducible reporter gene (mouse mammary tumor virus-luciferase). No effect was seen with PPARalpha- or hNUC1/PPARdelta-selective ligands. The GR antagonist RU-486 inhibited the DEX and TZD responses, suggesting that the effects were mediated through endogenous GR. TZD also enhanced glucocorticoid mediated transcription in SaOS-2/B10 human osteosarcomatous cells, but not in CV 1 cells, even though both cell lines were transfected with GR plasmid and expressed significant levels of endogenous PPARgamma messenger RNA. In MB 1.8 cells, TZD decreased alkaline phosphatase activity and the expression of osteoblast-associated genes while it up-regulated the adipocyte fatty acid binding protein. DEX counteracted the effects of TZD on alkaline phosphatase enzyme activity and osteoblastic gene expression, but enhanced the actions of TZD on adipocyte fatty acid-binding protein. Interestingly, TZD inhibited in vitro bone nodule formation and mineralization, and DEX counteracted this effect. Thus, depending on the promoter context, TZD and DEX can oppose or enhance each other's actions on gene transcription. Collectively, these results point to a complex interaction between PPAR and GR signaling pathways that regulates the effects of TZD and DEX on osteoblastic differentiation. The mechanism of this interaction is still under investigation, but might involve PPAR -dependent and -independent pathways. As thiazolidinediones represent an important new class of drugs, our findings also raise the need for further studies in bone. PMID- 10385422 TI - Fluctuating estrogen and progesterone receptor expression in brainstem norepinephrine neurons through the rat estrous cycle. AB - Norepinephrine (NE) neurons within the nucleus tractus solitarii (NTS; A2 neurons) and ventrolateral medulla (A1 neurons) represent gonadal steroid dependent components of several neural networks regulating reproduction. Previous studies have shown that both A1 and A2 neurons express estrogen receptors (ERs). Using double labeling immunocytochemistry we report here that substantial numbers of NE neurons located within the NTS express progesterone receptor (PR) immunoreactivity, whereas few PRs are found in ventrolateral medulla. The evaluation of ERa and PR immunoreactivity in NE neurons through the estrous cycle revealed a fluctuating pattern of expression for both receptors within the NTS. The percentage of A2 neurons expressing PR immunoreactivity was low on metestrus and diestrus (3-7%), but increased significantly to approximately 24% on proestrous morning and remained at intermediate levels until estrus. The pattern of ERalpha immunoreactivity in A2 neurons was more variable, but a similar increment from 11% to 40% of NE neurons expressing ERa was found from diestrus to proestrus. Experiments in ovariectomized, estrogen-treated and estrogen-plus progesterone-treated rats revealed that PR immunoreactivity in A2 neurons was induced strongly by estrogen treatment, whereas progesterone had no significant effect. The numbers of ERalpha-positive NE neurons were not influenced by steroid treatment. These observations provide direct evidence for PRs in NE neurons of the brainstem and show that cyclical patterns of gonadal steroid receptor expression exist in A2, but not A1, neurons through the rat estrous cycle. The expression of PR in A2 neurons appears to be driven principally by circulating estrogen concentrations. The fluctuating levels of ERalpha and PR expression in these brainstem NE neurons may help generate cyclical patterns of biosynthetic and electrical activity within reproductive neural networks. PMID- 10385423 TI - Involvement of protein kinase C and protein tyrosine kinase in thyrotropin releasing hormone-induced stimulation of alpha-melanocyte-stimulating hormone secretion in frog melanotrope cells. AB - We have previously shown that the stimulatory effect of TRH on alpha-MSH secretion from the frog pars intermedia is associated with Ca2+ influx through voltage-dependent Ca2+ channels, activation of a phospholipase C and mobilization of intracellular Ca2+ stores. The aim of the present study was to investigate the contribution of protein kinase C (PKC), adenylyl cyclase (AC), Ca2+/calmodulin dependent protein kinase II (CAM KII), phospholipase A2, and protein tyrosine kinase (PTK) in TRH-induced alpha-MSH release. Incubation of frog neurointermediate lobes (NILs) with phorbol 12-myristate-13-acetate (24 h), which causes desensitization of PKC, or with the PKC inhibitor NPC-15437, reduced by approximately 50% of the effect of TRH on alpha-MSH release. In most melanotrope cells, TRH induces a sustained and biphasic increase in cytosolic Ca2+ concentration ([Ca2+]i). Preincubation with phorbol 12-myristate-13-acetate or NPC-15437 suppressed the plateau phase of the Ca2+ response. Incubation of NILs with TRH (10(-6) M; 20 min) had no effect on cAMP production. In addition, the AC inhibitor SQ 22,536 did not affect the secretory response of NILs to TRH. These data indicate that the phospholipase C/PKC pathway, but not the AC/protein kinase A pathway, is involved in TRH-induced alpha-MSH release. The calmodulin inhibitor W-7 and the CAM KII inhibitor KN-93 did not significantly reduce the response to TRH. Similarly, the phospholipase A2 inhibitors quinacrine and 7-7'-DEA did not impair the effect of TRH on alpha-MSH secretion. The PTK inhibitors ST638 and Tyr A23 had no effect on TRH-induced [Ca2+]i increase but inhibited in a dose dependent manner TRH-evoked alpha-MSH release (ED50 = 1.22x10(-5) M and ED50 = 1.47x10(-5) M, respectively). Taken together, these data indicate that, in frog melanotrope cells, PKC and PTK are involved in TRH-induced alpha-MSH secretion. Activation of PKC is responsible for the sustained phase of the increase in [Ca2+]i, whereas activation of PTK does not affect Ca2+ mobilization. PMID- 10385424 TI - 3,5 cyclic adenosine monophosphate mediates the salmon calcitonin-induced increase in hypothalamic tyrosine hydroxylase activity. AB - This study examined the effect of salmon calcitonin (sCT) on hypothalamic tyrosine hydroxylase (TH) activity and evaluated the cellular signaling mechanisms involved in the response. Fetal hypothalamic cells were cultured in a defined medium and treated with sCT and/or specific protein kinase inhibitors on day 14 in vitro. sCT (0.1-10 nM) increased both TH activity and cellular cAMP content in a concentration-dependent manner. sCT (10 nM) increased TH activity to 150-175% of control values and resulted in a 10-fold increase in cellular cAMP content. Both the C1a and C1b CT receptor isoforms were present in the cultures, as assessed by RT-PCR. Rp-adenosine 3',5'-cyclic monophosphothioate (Rp-cAMPS), a cAMP antagonist, and H-8, a cyclic nucleotide kinase inhibitor, blocked the sCT induced increase in TH activity, with complete abolition of the response observed at concentrations of 1 mM and 5 microM, respectively. sCT (10 nM) increased radiolabeled phosphate incorporation into TH protein to 169% of control values and 1 mM Rp-cAMPS completely blocked this effect. In contrast, neither Calphostin C, a protein kinase C inhibitor, nor U-73122, a phospholipase C inhibitor, significantly altered the ability of sCT to increase TH activity. Likewise, the sCT-induced increase in TH activity was observed after pretreating the cells with either BAPTA/AM, an intracellular calcium chelator, or thapsigargin, an inhibitor of the endoplasmic reticulum calcium pump. These data indicate that sCT has a profound stimulatory effect on TH activity in fetal hypothalamic cells and that enhanced phosphorylation of TH coincides with the sCT-induced increase in enzyme activity. Moreover, CT receptors, which are linked to cAMP production, are expressed in the hypothalamic cells and a cAMP-dependent mechanism mediates the sCT-induced activation and phosphorylation of TH. PMID- 10385425 TI - Differential posttranscriptional regulation of androgen receptor gene expression by androgen in prostate and breast cancer cells. AB - Androgens, via the androgen receptor (AR), modulate the growth and proliferation of prostate and breast cancer cells. However, the molecular mechanisms underlying the regulation of AR gene expression by androgen in these cells remain to be fully elucidated. To explore differences in AR gene expression between these hormone-responsive tumor cell types, we studied androgen-responsive LNCaP prostate cancer and AR positive MDA453 breast cancer cells. Dihydrotestosterone (DHT) 10 nM increased LNCaP cell proliferation and the proportion of LNCaP cells in S-phase of the cell cycle but inhibited MDA453 cell proliferation and reduced the proportion of MDA453 cells in S-phase of cell cycle. In both these cell lines, DHT decreased total AR messenger RNA (mRNA) but increased AR protein. In LNCaP cells, DHT down-regulated AR mRNA transcription but stabilized AR mRNA. In contrast, in MDA453 cells, DHT had no effect on AR mRNA transcription but destabilized AR mRNA. In summary, transcriptional down-regulation induced by androgens in LNCaP cells results in down-regulation of steady-state AR mRNA despite an androgen-induced increase in AR mRNA stability. However, in MDA453 cells, posttranscriptional destabilization of AR mRNA appears to be the predominant mechanism resulting in down-regulation of AR mRNA by androgen. These results demonstrate cell-specific and divergent regulation of AR mRNA turnover by androgen and identify a novel pathway of androgen-induced posttranscriptional destabilization and down-regulation of AR mRNA in human breast cancer cells. Furthermore, these data establish an important role for posttranscriptional pathways in the regulation of AR gene expression by androgen in human prostate and breast cancer cells. PMID- 10385426 TI - The monkey and human uridine diphosphate-glucuronosyltransferase UGT1A9, expressed in steroid target tissues, are estrogen-conjugating enzymes. AB - Considering the physiologic importance of the steroid response, which is regulated in part by steroid levels in a given tissue, relatively little is known about steroid glucuronidation, which is widely accepted as a major pathway involved in the catabolism and elimination of steroid hormones from the human body. In a previous study, it was ascertained that the monkey may be the most appropriate model in which to examine the role of steroid glucuronidation. Northern blot analysis of simian RNA, hybridized with human UGT complementary DNA (cDNA) probes demonstrate the similarity of the transcripts. The simian UGT1A09 cDNA isolated from a liver library is 2396 bp and contains an open reading frame encoding 530 amino acids. The predicted primary structure is most homologous to the human UGT1A9 (hUGT1A9) enzyme, which share 93% identity. Stable transfection of the monkey UGT1A09 (monUGT1A09) cDNA into HK293 cells, expresses a microsomal protein with an apparent molecular mass of 55 kDa. Of the more than 30 endogenous substrates tested, both proteins show the highest activity on 4-hydroxyestradiol and 4-hydroxyestrone, followed by 2-hydroxyestradiol and estradiol. RT-PCR analysis demonstrate that UGT1A9 transcript is expressed in several tissues, which include the prostate, testis, breast, ovary, and skin of the monkey and humans. The expression of UGT1A9 in extrahepatic estrogen-responsive tissues, and its high activity on estrogens is consistent with this enzyme having a role in estrogen metabolism. PMID- 10385427 TI - Expression and activity of vitamin D-metabolizing cytochrome P450s (CYP1alpha and CYP24) in human nonsmall cell lung carcinomas. AB - Extrarenal 25-hydroxyvitamin D3-1alpha-hydroxylase is believed to play a major role in the pathogenesis of hypercalcemia associated with various types of granulomatous and lymphoproliferative diseases and certain solid tumors. In this paper, we describe the cloning of the cytochrome P450 component of the extrarenal enzyme from a human nonsmall cell lung carcinoma, SW 900. The cytochrome P450 for the extrarenal 1alpha-hydroxylase has an amino acid sequence identical to that of the cytochrome P450 component of the CYP1alpha, the renal form of the enzyme, and appears to be a product of the same gene. CYP1alpha messenger RNA (mRNA) and 1alpha-hydroxylase enzyme activity were detected in two (SW 900, SK-Luci-6) of a series of five nonsmall cell lung carcinoma cell lines. All five lung cell lines were cultured with the same medium under the same conditions, but only two of the five expressed 1alpha-hydroxylase enzyme; two others (WT-E, Calu-1) expressed high levels of the reciprocally regulated enzyme, 25-hydroxyvitamin D3-24 hydroxylase, with its specific cytochrome P450 component, CYP24. Although under basal conditions the lung cell line SW 900 expressed only CYP1alpha and showed 1alpha-hydroxylase enzyme activity, when treated with small concentrations of 1alpha,25-dihydroxyvitamin D3 or high concentrations of 25-hydroxyvitamin D3, it began to express CYP24 and exhibit 24-hydroxylase enzyme activity. Somewhat surprisingly, SW 900 cells still had detectable CYP1alpha mRNA some 24 h after vitamin D treatment despite the fact that 1alpha-hydroxylase enzyme activity was unmeasurable. These data are consistent with the emerging hypothesis that vitamin D through its active form does not directly turn off CYP1alpha mRNA production but, rather, strongly stimulates CYP24, thereby masking CYP1alpha activity. The factor(s) responsible for the basal expression of CYP1alpha in SW 900 and SK-Luci 6 is currently unknown. PMID- 10385428 TI - Increase in PDX-1 levels suppresses insulin gene expression in RIN 1046-38 cells. AB - RIN1046-38 cells (RIN-38) exhibit a passage-dependent reduction in both basal and glucose-regulated insulin secretion, accompanied by decreased insulin content. In an attempt to explain the mechanism of the gradual decrease in insulin production in cultured cells, we analyzed the insulin promoter activity and the levels of an important trans-activator of the insulin gene, PDX-1, as a function of aging in culture. We demonstrate that the decrease in insulin content and secretion is reflected in decreased promoter activity and is associated with a decrease in E47 and BETA2 nuclear factors, but with a paradoxical 3-fold increase in PDX-1 protein levels. To dissect the effect of increased PDX-1 from the decrease in the additional transcription factors on insulin promoter activity, we overexpressed PDX-1 protein in low passage RIN-38 cells by recombinant adenovirus technology. PDX-1 overexpression did not reduce E47 and BETA2 levels, but was sufficient to suppress rat insulin promoter activity in a dose-dependent manner. The fact that PDX-1 levels participate in trans-activation of insulin promoter activity was demonstrated in HIT-T15 cells. Treating HIT-T15 cells with 1-2 multiplicity of infection of AdCMV-PDX-1 increased rat insulin promoter activity, whereas higher doses repressed insulin promoter activity in these cells as in RIN-38 cells. Our data demonstrate that PDX-1 regulates transcription of the insulin gene in a dose dependent manner. Depending on its nuclear dosage and the levels of additional cooperating transcription factors, PDX-1 may act as an activator or a repressor of insulin gene expression, such that low as well as high doses may be deleterious to insulin production. PMID- 10385429 TI - P34H sperm protein is preferentially expressed by the human corpus epididymidis. AB - During epididymal transit, mammalian spermatozoa acquire new surface proteins that are necessary for gamete interaction. We have previously described a 34-kDa human epididymal sperm protein, P34H, that has been shown to be involved in sperm zona pellucida interaction. In the present study, we report the cloning and characterization of the full-length complementary DNA encoding human P34H. The predicted amino acid sequence revealed 65% identity with P26h, the hamster counterpart of the P34H. The deduced P34H amino acid sequence revealed a 71% similarity with a pig lung tetrameric carbonyl reductase, a member of the short chain dehydrogenase/ reductase family proteins. Northern blot analysis revealed that P34H messenger RNA (mRNA) was highly expressed in the human epididymis, principally in the corpus region. A single 912-bp P34H transcript was detected. In situ hybridization experiments showed that the P34H mRNA was predominantly expressed in the proximal and distal sections of the corpus epididymidis. The staining was restricted to the principal cells of the epididymal epithelium. The localization of P34H mRNA was in agreement with the appearance of P34H protein along the male reproductive tract. Western blot analysis revealed that recombinant P34H expressed by a yeast expression system, is antigenically related to the native P34H sperm protein. Based on its pattern of expression and its function in one of the key steps leading to fertilization, P34H can be considered as a marker of epididymal sperm maturation in humans. PMID- 10385430 TI - Expression of p190A during apoptosis in the regressing rat ventral prostate. AB - After hormonal ablation, 90% of the secretory epithelial cells of the prostate undergo apoptosis, and the remaining cells are reorganized as the tissue is remodeled. Using differential display RT-PCR of total RNA extracted from the rat ventral prostate before and 4 days after castration, we have cloned and sequenced a number of complementary DNAs whose cognate messenger RNAs (mRNAs) may be either up- or down-regulated during prostatic regression. One sequence of particular interest, 25.2, is up-regulated after castration and is homologous to p190, a protein associated with cytoskeletal reorganization. RT-PCR has confirmed that the steady state level of p190A mRNA is increased in the rat ventral prostate after castration, and Western blot analysis indicates that the protein levels for p190A also increase. The steady state level of p190B mRNA, the second isoform of p190, does not appear to change significantly after hormone ablation. Immunohistochemical analysis demonstrates that p190A is up-regulated primarily in the columnar epithelial cells that actively undergo cell death after hormone ablation. As Rho-GAP signaling had been shown to be influenced by p190 levels, leading to the disassembly of focal adhesion contacts and the loss of cytoskeletal architecture, we also measured the changes in Rho-GAP during prostate regression. Rho-GAP levels do not change significantly, suggesting that changes in stoichiometry of the interaction between p190A and Rho-GAP may be a prerequisite for the initiation of cytoplasmic condensation. These intracellular events coupled with the proteolytic degradation of the extracellular matrix appear to be integral to the apoptotic process in glandular epithelia. PMID- 10385431 TI - Characterization of molecular and catalytic properties of intact and truncated human 17beta-hydroxysteroid dehydrogenase type 2 enzymes: intracellular localization of the wild-type enzyme in the endoplasmic reticulum. AB - Human 17beta-hydroxysteroid dehydrogenase (17HSD) type 2 is a widely distributed enzyme that primarily converts the highly active 17beta-hydroxysteroids to their inactive keto forms. In the present study, full-length human 17HSD type 2 was localized in the endoplasmic reticulum using a double immunofluorescence labeling technique. As a consequence of its strong membrane interaction, full-length human 17HSD type 2 could not be solubilized as a biologically active form in vitro. However, by deleting the first 29 amino acids from the N-terminus, we were able to purify a catalytically active enzyme from the cytosolic fraction of Sf9 insect cells. Biochemical and catalytic properties of the purified truncated human 17HSD type 2 protein confirm its suitability for structure-function analyses of the enzyme. Both intact and truncated 17HSD type 2 enzymes efficiently catalyzed the oxidation of estradiol, testosterone, dihydrotestosterone, androstenediol, and 20alpha-dihydroprogesterone. The oxidation of estradiol brought about by human 17HSD type 2 was effectively inhibited by several other steroidal compounds, such as 2-hydroxyestradiol, 5beta-androstan-3alpha,17beta-diol, 5alpha-androstan 3alpha,17beta-diol, and 5alpha-androstan-3beta,17beta-diol. The broad substrate specificity of human 17HSD type 2 together with its predominant oxidative activity and intracellular location, as observed in this study, indicate the physiological role of the enzyme to be primarily an inactivator of highly active 17beta-hydroxysteroids. PMID- 10385432 TI - Development of functional zonation in the rat adrenal cortex. AB - In an attempt to elucidate the mechanism(s) through which the functional adrenal cortex is established, we analyzed immunohistochemically the expression of various markers for the adrenocortical zones, i.e. the zona glomerulosa (zG), the zona fasciculata (zF), and the zona reticularis (zR), as well as markers for the medulla, and further examined the distribution and behavior of DNA-synthesizing cells in rat adrenal glands during development. The results showed that 1) separation of the cortex and medulla, and the development of functional zonation in the cortex began at around the time of birth, 2) at fetal stages when cortical zonation was not established, DNA-synthesizing cells were found scattered throughout the gland, where they proliferated without significant migration, and 3) after birth in the adrenal cortex with established cortical zonation, DNA synthesizing cells were localized near the undifferentiated zone between zG and zF, and then they migrated centripetally. Cell death appeared to occur in the innermost portion of the cortex, where many resident macrophages are present. These findings illustrate basic processes underlying adrenal development and suggest that the undifferentiated region is apparently the stem cell zone of the adrenal cortex that maintains the cortical zonation. PMID- 10385433 TI - Prenatal dexamethasone treatment does not prevent alterations of the hypothalamic pituitary adrenal axis in steroid 21-hydroxylase deficient mice. AB - A major difficulty in the clinical management of congenital adrenal hyperplasia (CAH) is adjustment of glucocorticoid doses to suppress ACTH and androgens without causing iatrogenic hypercortisolism. The possibility that structural alterations of the adrenal or a dysfunction of the hypothalamic pituitary adrenal (HPA) axis caused by glucocorticoid deficiency during fetal life contribute to this problem was studied in 21-hydroxylase deficient mice caused by deletion of the cytochrome P-450 21-hydroxylase gene. Homozygotes showed about 200-fold elevations in plasma progesterone, hyperplastic adrenal cortices lacking zonation, and structural alterations of adrenocortical mitochondria. Histochemical studies showed increases in hypothalamic CRH messenger RNA (mRNA) and immunoreactive (ir) CRH, and pituitary POMC mRNA in homozygous mice. VP mRNA levels in PVN perikarya were normal, but irVP in parvicellular terminals of the median eminence was increased in homozygotes. Prenatal dexamethasone treatment (0.5 to 2 microg/day) prevented the increases in CRH mRNA, whereas dexamethasone only partially decreased POMC mRNA levels, and had no effect on serum progesterone levels. The data suggest that intrauterine glucocorticoid deficiency in CAH causes hyperactivity of the hypothalamic-pituitary-corticotroph axis and insensitivity to glucocorticoid feedback. These studies in 21-hydroxylase deficient mice may provide new insights on the mechanism, clinical manifestations and management of some types of human CAH. PMID- 10385434 TI - Distribution of the parathyroid hormone 2 receptor in rat: immunolocalization reveals expression by several endocrine cells. AB - The PTH2 receptor is a G protein-coupled receptor selectively activated by PTH. We are studying the receptors distribution to guide the investigation of its physiological function. We have now generated an antibody from a C-terminal peptide sequence of the PTH2 receptor and used this to study its cellular distribution. Labeling with the antibody identified a number of endocrine cells expressing the PTH2 receptor, including thyroid parafollicular cells, pancreatic islet D cells, and some gastrointestinal peptide synthesizing cells. There was complete overlap of PTH2 receptor labeling with somatostatin in pancreatic islets, and partial overlap with somatostatin in thyroid parafollicular cells and in the gastrointestinal tract. Furthermore, observations made previously by in situ hybridization histochemistry, including expression throughout the cardiovascular system, as well as by discrete populations of cells within the gastrointestinal tract and reproductive system were confirmed. These data suggest a broad role for the PTH2 receptor, especially within the endocrine system, and provide a basis for experimental exploration of its physiology. PMID- 10385435 TI - Localization of bradykinin B2 receptors in the endometrium and myometrium of rat uterus and the effects of estrogen and progesterone. AB - In the uterus, bradykinin is a potent inducer of smooth muscle contraction, which is mediated by the bradykinin B2 receptor subtype. However, little is known about the distribution or regulation of this receptor in this tissue. The aim of this study was to localize the B2 receptor in the uterus and determine whether the levels of this receptor were altered during the estrous cycle and modulated by estrogen and/or progesterone in ovariectomized rats. At diestrus, uterine B2 receptors were localized to both the circular and longitudinal smooth muscle layers of the myometrium, the endometrial stroma, the glandular epithelium, and the layer subjacent to the luminal epithelium. B2 receptor levels in both myometrium and endometrium were lowest during early proestrus, when estrogen levels are low, whereas myometrial B2 receptor protein and messenger RNA levels were highest during late proestrous, when estrogen levels peak. Similar findings were observed for the estrogen-supplemented group after ovariectomy, with progesterone appearing to inhibit the estrogen-induced rise in bradykinin B2 receptor density in estrogen/progesterone-treated animals. Using in vitro receptor autoradiography employing the specific B2 receptor antagonist analog, HPP-HOE140, immunostaining with specific antipeptide antibodies generated against the B2 receptor, and in situ hybridization using a specific bradykinin B2 receptor riboprobe, our findings show a discrete distribution of the bradykinin B2 receptor throughout the different layers of the uterus and suggest that bradykinin B2 receptor levels in the rat uterus are regulated by estrogen, and possibly progesterone, in both myometrium and endometrium. PMID- 10385436 TI - Androstenedione effects on the vasopressin innervation of the rat brain. AB - The steroid hormone androstenedione profoundly influences the development and expression of sexual and aggressive behavior. The neural basis of these effects are, however, poorly understood. In this study we evaluated androstenedione's ability to maintain vasopressin peptide levels in the gonadal steroid-responsive vasopressin cells of the bed nucleus of the stria terminalis and the centromedial amygdala, and their projections. Adult male rats were castrated and given testosterone, androstenedione or no hormonal treatment for five weeks. Their brains were then processed for vasopressin immunoreactivity. Androstenedione and testosterone treatment were equally effective in preventing the reduction of vasopressin immunoreactivity associated with castration. Androstenedione may therefore be able to mimic the effects of testosterone on testosterone-responsive neural systems. PMID- 10385437 TI - Recombinant E-peptides of pro-IGF-I have mitogenic activity. AB - In mammals, the post-translational proteolytic modification of many pro-hormones generates multiple peptides with similar or distinct biological activities. The production of mature insulin-like growth factor I (IGF-I) involves the cleavage of an E-peptide from pro-IGF-I. Although the IGF-I E-peptides are conserved among vertebrate species, their fate and biological roles have not been identified. To test whether the E-peptides possess any biological activity, three recombinant E peptides of pro-IGF-I, namely Ea-2-, Ea-3- and Ea-4-peptides of rainbow trout, Oncorhynchus mykiss, were produced in vitro with a His-tag expression system and partially purified with an affinity Ni++ column. The mitogenic activity of each E peptide was determined by (1) the stimulation of total DNA content increase as measured by a fluorometric method and/or (2) stimulation of [3H]-thymidine incorporation into DNA. Recombinant Ea-4-peptide elicited a dose-related increase in both mitogenic assays in NIH3T3 cells. To further test the specificity of the mitogenic activity of Ea-4-peptide, three other cell lines were used: retroviral transformed human embryonic kidney cells (293GP), human mammary gland tumor cells (MCF-7) and caprine mammary epithelial cells (CMEC). Similar levels of mitogenic activity were observed in all cell lines tested for Ea-4-peptide. Mitogenic activity was also observed with recombinant Ea-2- and Ea-3-peptides when assayed in NIH3T3 cells. These results suggest that E-peptides of rainbow trout pro-IGF-I possess mitogenic activity in heterologous systems. PMID- 10385438 TI - Effects of Igf1 gene deletion on postnatal growth patterns. AB - This study documents the temporal and organ-specific effects of Igf1 gene deletion on postnatal growth patterns. Igf1-/- mice are 63+/-4% the size of wildtype (wt) littermates at birth and this ratio persists through postnatal day 20 (P20). After P20, Igf1-/- mice virtually stop growing, while wt littermates double in size from P20 to P40, after which their growth markedly decelerates. As a result, 'full-grown' Igf1-/- mice are less than one third the size of wt littermates. Igf1 gene deletion has disproportionate effects on organ growth. For example, at P10 and P40, Igf1-/- body weights are 63% and 31% of wt, respectively, while Igf1-/- lungs weigh only 34% and 22% of wt at these ages. In contrast, the Igf1-/- heart is disproportionately enlarged, representing approximately 85% of wt at P10 and approximately 56% at P40. Igf1-/- kidney, spleen and liver are slightly but significantly increased in size relative to the degree of reduction in Igf1-/- body weight. These data demonstrate that Igf1 has two major phases or modes of growth promotion. There is an early, growth hormone (GH)-independent Igf1 growth augmentation during perinatal development, responsible for about 35% of growth prior to P20. Then there are later effects due to GH-induced Igf1, which are responsible for increasing animal size by approximately 100% between P20 and 40. The fact that there is virtually no GH induced growth in the Igf1-/- mice supports the view that Igf1 mediates GH's major effects on somatic growth. Finally, this study shows that Igf1 has discordant effects on pulmonary and cardiac growth parameters, with relative hypoplasia of Igf1-/- lungs and hypertrophy of Igf1-/- hearts. PMID- 10385439 TI - Reflections on adequacy in cervical/vaginal cytology. PMID- 10385440 TI - ThinPrep Pap Test: performance and biopsy follow-up in a university hospital. AB - BACKGROUND: The ThinPrep Pap Test (TP), a liquid-based cervical cytology preparation, was approved for use in the U.S. in 1996. The purpose of this study was to compare TP performance and biopsy follow-up studies with a similar population of high risk patients sampled by conventional Papanicolaou (Pap) smear (CS). METHODS: Diagnostic and specimen adequacy interpretations for 2727 TP direct-to-vial Pap tests from a high risk university hospital practice were compared with 5000 CS preparations from the same physicians taken 1 year previously. Biopsy follow-up studies for the categories of squamous intraepithelial lesion (SIL), carcinoma, and atypical squamous cells of undetermined significance (ASCUS) for each time period and technique were contrasted. RESULTS: The SIL/carcinoma detection rate increased from 7.7% to 10.5% (P < 0.01) and the ASCUS rate decreased from 12.5% to 6.9% (P < 0.01); the percentage of satisfactory but limited specimens decreased from 19.4% to 10.5% (P < 0.01). Low grade SIL cases increased by 57% (P < 0.01) whereas the 26% increase in high grade SIL cases was not statistically significant. Greater than 90% of ungraded SIL, high grade SIL, and carcinoma cases had abnormal biopsies by both the TP and CS methods. The number of biopsy-confirmed high grade dysplasias and carcinomas was similar in the two groups. A low grade SIL detected by TP was less likely to have an abnormal biopsy (70% vs. 85% for CS). Nevertheless, the 57% increase in low grade SIL diagnoses by TP resulted in more TP patients with dysplastic biopsy diagnoses. Follow-up studies for ASCUS cases diagnosed by either TP or CS were similar, and 21-24% of patients eventually were found to have dysplasia. CONCLUSIONS: The TP technique appears to lead to the increased detection of low grade SIL lesions, decreased satisfactory but limited samples, and fewer equivocal specimens. No increase in biopsy-confirmed high grade dysplasias and carcinomas was found. Follow-up studies for the ASCUS category were nearly identical to those for CS. PMID- 10385442 TI - The accuracy of urinary cytology in daily practice. AB - BACKGROUND: Most studies of urinary cytology have been research analyses designed to test the method itself, and many claim that the high diagnostic yields in these studies cannot be achieved in daily practice. The authors examined the clinical and pathologic records in three hospital pathology practice settings- academic, community, and cancer referral settings--to determine the diagnostic yield of urinary cytology under daily clinical conditions. METHODS: Records of consecutive urinary cytology specimens from 1672 patients reported from the years 1990-1994 were reviewed and correlated with histologic and clinical information. Initial analyses were based on the records themselves, without review of pathologic specimens. Subsequently, a subset of specimens was reviewed to determine reasons for noncorrelations. RESULTS: Results confirmed that the diagnostic sensitivity and specificity of urinary cytology for high grade transitional cell neoplasms, as reported in the daily practice of pathology, are very high (79% and >95%, respectively). Disaggregated cells from low grade transitional cell neoplasms usually lack recognizable features of neoplasia, and attempts to classify such lesions cytologically result in low diagnostic yield, with an overall sensitivity of 26%. Of these 1672 patients, 707 had insufficient clinical information for analysis, despite diligent and persistent efforts to acquire it. CONCLUSIONS: The diagnostic yield of consultations based on urinary cytology in the daily practice of pathology is high, regardless of whether the practice setting is referral-based or community-based. The available information indicates that in approximately 79% of patients with high grade transitional cell neoplasms, the neoplasms can be detected using urinary cytology. Conversely, a negative result is predictive of no cancer in more than 90% of cases. Sensitivity for detecting low grade urothelial lesions is low; however, most of these are easily detected cystoscopically. The authors' inability to acquire sufficient information to determine diagnostic yield in a large percentage of their cases was disturbing to them. Not only did this deficiency render their analyses incomplete, but lack of easily accessible follow-up and the apparent low priority for quality assurance activities among pathologists in all types of practice settings is likely to significantly reduce the feedback required for pathologists to acquire and maintain expertise in this very difficult area. PMID- 10385441 TI - The effect of the quality of Papanicolaou smears on the detection of cytologic abnormalities. AB - BACKGROUND: Controversy continues regarding the relation between the quality of Papanicolaou (Pap) smears, especially the presence of endocervical cells (ECC), with the finding of cytologic abnormalities. METHODS: As part of a study regarding performance feedback on the quality of Pap smears, data from 56,475 Pap smears obtained by 176 participating clinicians over a 20-month period were analyzed to assess the relation between the presence of ECC, the categorization of global specimen adequacy as "satisfactory" or "satisfactory with limitations," and the prevalence of atypia and squamous intraepithelial lesions (SILs). RESULTS: Atypia was less likely to be found in "satisfactory" Pap smears than in "satisfactory with limitations" quality Pap smears (odds ratio [OR], 0.6; 95% confidence interval [CI], 0.5-0.6; P < 0.001), even though the latter could contain ECC. No association was found between satisfactory Pap smears and cytologic abnormalities. Compared with specimens with no ECC, an ECC count of > or = 50 on a slide was associated positively with the detection of atypia (OR, 2.1; 95% CI, 1.8-2.4; P < 0.001) or SILs (OR, 1.7; 95% CI, 1.3-2.2; P < 0.001). A similar relation existed between ECC counts of 25-50 (OR, 1.9; 95% CI, 1.1-2.2; P = 0.01) and the detection of SILs. No relation was found between specimens with < 25 ECC and the presence of atypia or abnormalities. CONCLUSIONS: The global adequacy criterion of "satisfactory" assigned to a Pap smear does not indicate that there is a greater likelihood of detecting cytologic abnormalities compared with lower quality Pap smears. To the authors' knowledge, previous studies regarding the link between ECC in the Pap smear and cytologic abnormalities have not addressed the relevance of how many ECC are needed to maximize the identification of abnormalities. The data from the current study support the value of obtaining at least 25 ECC as a quality indicator of sampling. PMID- 10385443 TI - A prospective comparison of fiberoptic transbronchial needle aspiration and bronchial biopsy for bronchoscopically visible lung carcinoma. AB - BACKGROUND: Fiberoptic bronchoscopy is the most common modality used to diagnose endobronchial carcinoma. The authors prospectively compared the sensitivity of endobronchial needle aspiration (EBNA) and immediate cytologic assessment with bronchial biopsy and bronchial washing in the diagnosis of endobronchial malignancy. METHODS: A prospective trial comparing the sensitivity of EBNA, bronchial biopsy, and bronchial washings during fiberoptic bronchoscopy for endobronchially visible lung tumor was conducted. The authors enrolled 65 consecutive patients with endobronchial abnormalities identified during bronchoscopy. All patients in the study underwent fiberoptic bronchoscopy that included EBNA, bronchial biopsy, and bronchial wash. The sensitivities of the individual techniques were compared. The sensitivities of bronchoscopy were also prospectively compared when multiple sampling techniques were employed. RESULTS: Malignancy was present in 57 of 65 study patients. Cancer was diagnosed in 47 patients by EBNA, 42 patients by bronchial biopsy, and 36 patients by bronchial washing. The sensitivity of a strategy employing bronchial biopsy and bronchial washings was 0.82 (95% CI, 0.70-0.90). The addition of EBNA to bronchial biopsy and bronchial washings significantly increased the sensitivity to 0.95 (95% CI, 0.85-0.98; McNemar P = 0.02). Subset analysis revealed that this strategy was especially useful in cases in which lesions were submucosal or causing extrinsic compression. CONCLUSIONS: There is a modest increase in the sensitivity of fiberoptic bronchoscopy in diagnosing endobronchial cancer with the addition of EBNA to bronchial biopsy and bronchial washings, especially for patients with submucosal abnormalities. Collection of EBNA, followed by biopsy and washings only if immediate interpretation of EBNA is negative or inadequate, may be the most effective bronchoscopy strategy for evaluating visible endobronchial abnormalities. PMID- 10385444 TI - Cytologic analysis of renal angiomyolipoma: a comparison of radiologically classic and challenging cases. AB - BACKGROUND: Angiomyolipoma is a benign kidney tumor with distinctive pathologic and clinical features. Because the prognosis differs from many other renal and retroperitoneal tumors, accurate diagnosis on cytologic material may be important for appropriate management. METHODS: A retrospective analysis of cytologic material from eight patients with histologically confirmed angiomyolipomas was performed. The findings are described. RESULTS: Three cases were diagnosed radiologically as angiomyolipoma, three were diagnosed as renal cell carcinoma, one was diagnosed as a "renal mass," and one was diagnosed as a fat-containing adrenal tumor. Compared with the tumors that were suggestive radiologically of angiomyolipoma, tumors that were suspicious radiologically for renal cell carcinoma tended to contain less fat on cytologic examination. One fat-containing tumor was not diagnosed as angiomyolipoma because the tumor was believed to be adrenal in origin. Cytologically, cohesive stromal fragments were comprised of cells ranging from epithelioid to elongate. Stromal atypia was present in 7 of 8 cases (88%) but was marked in only 2 cases (25%). Thick-walled vessels were noted in 3 of 8 cases (38%). CONCLUSIONS: The cytologic diagnosis of angiomyolipoma is difficult in cases in which the radiologic diagnosis is not clear. The liberal use of immunostains is advised in the evaluation of stromal renal tumors. PMID- 10385445 TI - The cytology of extraskeletal Ewing sarcoma. AB - BACKGROUND: Extraskeletal Ewing sarcoma (EES) shares histologic, immunohistochemical, and molecular findings with ES of bone. The authors' goal in conducting this study was to examine the cytomorphologic features of EES. In addition, immunocytostaining for CD99/O13 was performed in all cases, and cytogenetic and molecular data were available in about half of the cases. METHODS: The authors studied 20 aspiration cases, all with histopathologic confirmation, and also conducted immunohistochemistry and/or molecular studies. RESULTS: All cases had cellular smears with many single cells and focal clustering. Numerous naked nuclei and focal crush artifacts were seen. Mitosis and necrosis were rare. Four cases had cytoplasmic vacuoles. Five cases showed nuclear molding. Seventeen cases (85%) exhibited small cells with scanty cytoplasm and nuclei with fine chromatin and small nucleoli, representing the so called typical variant. One case (5%) revealed cells with abundant cytoplasm, large nuclei, and large eosinophilic nucleoli, an example of the atypical or large cell variant. Two cases (10%) had features in between, with cells showing a fair amount of cytoplasm and medium-sized nucleoli, representing the intermediate variant. Nuclear grooves, described as common in the latter, were rare. In all cases, in either cytologic or corresponding histologic material, CD99/O13 immunocytostaining showed strong membranous reactivity. In addition, cytogenetic and/or molecular evidence of ES specific chromosomal translocation was demonstrated in histologic or cytologic material in 10 cases. CONCLUSIONS: EES shows cytologic features similar to ES of bone, with a spectrum of changes ranging from the typical appearance in a majority of cases to intermediate and atypical variants in a minority of cases. CD99/O13 immunocytostaining and/or molecular studies, particularly in the intermediate and atypical variants, may help in establishing a definitive fine-needle aspiration diagnosis, thus avoiding an open surgical biopsy. PMID- 10385446 TI - Utility of punch biopsy for lesions that are hard to aspirate by conventional fine-needle aspiration. AB - BACKGROUND: Fine-needle aspiration (FNA) is a fast, reliable, and cost-efficient technique for diagnosing palpable masses. However, when the lesion is small, dermal in location, shallow in depth, or fibrotic, the cellular yield by FNA may be limited and thus hinder an accurate diagnosis. The authors examined the value of punch biopsy (PB) in diagnosing such hard-to-aspirate lesions. METHODS: The authors reviewed 49 PB specimens from 47 patients who presented in their FNA clinic from June 1994 to July 1997. RESULTS: The lesions were typically described as ill-defined erythematous skin lesions or as papules or small, firm, subcutaneous nodules (average size, 0.7 cm). Patients' previous history included breast carcinoma (in 36 cases), nonmammary carcinoma (in 3 cases), melanoma (in 2 cases), squamous carcinoma of the skin (in 2 cases), cutaneous T-cell lymphoma (in 2 cases), small lymphocytic lymphoma (in 1 case), and no history of malignancy (in 1 case). PB sites included chest wall, breast, extremities, abdominal wall, neck, back, scalp, and forehead. Of the 37 cases in which FNA was performed before PB, 21 aspirates (57%) were nondiagnostic because of scant cellularity, 11 aspirates (31%) were positive (9) or suspicious/atypical (2) for malignancy, and 5 aspirates (14%) were negative for malignancy. Seventeen (81%) of the 21 nondiagnostic aspirates and 10 of the 11 suspicious/atypical aspirates were positive for malignancy on PB specimens. Twelve PBs were done without prior FNA, 8 (67%) were positive for malignancy, and 4 (33%) were negative. In 7 patients, the findings from the PB specimens (new diagnosis of malignancy in 5 cases and recurrence of disease in 2 cases) led to surgical excision of the lesion. CONCLUSIONS: PB is a valuable adjunct to FNA for diagnosing hard-to aspirate lesions. PMID- 10385447 TI - Image analysis of papillary thyroid carcinoma fine-needle aspirates: significant association between aneuploidy and death from disease. AB - BACKGROUND: Papillary thyroid carcinoma is the most common thyroid malignancy in the U.S. As many as half of patients with papillary carcinoma present with cervical lymph node metastases at the time of diagnosis. Metastatic disease involving cervical lymph node tissue has not historically been proven to correlate with a more aggressive course; however, distant metastases worsen prognosis. METHODS: Diagnostic fine-needle aspiration (FNA) smears from 26 primary and metastatic papillary carcinomas underwent Feulgen reaction and were studied by image analysis to determine DNA pattern, proliferation index, and the percentage of cells with DNA content >5C. The medical records of all the patients were reviewed for metastatic disease pattern and survival data. For metastatic pattern, two groups were defined: 1) confined to thyroid/local lymph node metastases/soft tissues of the neck involved by tumor, and 2) distant metastases. RESULTS: Among the 26 patients, 16 had "nonaggressive" DNA patterns described as diploid, abnormal diploid, or tetraploid, and 10 had "aggressive" DNA patterns described as aneuploid. Only 2 of the 16 patients in the "nonaggressive" DNA pattern group developed distant metastases, whereas 5 of the 10 patients in the aneuploid group developed distant metastatic disease. In addition, none of the 16 patients with "nonaggressive" DNA patterns died of disease, whereas 3 of the 10 individuals with DNA histograms interpreted as aneuploid did die of metastatic disease complications. CONCLUSIONS: Aneuploidy identified by image analysis of FNA of papillary thyroid carcinoma is significantly associated with death from papillary carcinoma (log rank test, P=0.027). PMID- 10385448 TI - Fine-needle aspiration biopsy of chromophobe renal cell carcinoma and oncocytoma: comparison of cytomorphologic features. AB - BACKGROUND: Chromophobe renal cell carcinoma (ChRCC) is a distinct tumor with a prognosis intermediate between renal oncocytoma (RO) and clear cell renal cell carcinoma. To our knowledge the cytologic features of only a limited number of ChRCC have been described to date. A retrospective review of the cytomorphologic features of ChRCC and a comparison with RO was performed. METHODS: Fine-needle aspiration biopsies (FNABs) of six cases of histopathologically proven ChRCC were reviewed. The material examined was comprised of smears, cytospins, Thin Prep Pap Test preparations, and cell block sections stained with Diff-Quik, Papanicolaou, and hematoxylin and eosin. Six FNABs of ROs were examined similarly. The cytomorphology of each tumor was studied and particular attention was paid to features differentiating the two entities. RESULTS: The characteristic cytomorphologic features of ChRCC (present in all cases) included round/oval, occasionally polygonal, moderately pleomorphic large cells present singly and in small clusters. The abundant cytoplasm was variegated, ranging from dense to flocculent to vacuolated, with prominent cytoplasmic membranes. The nuclei were large and hyperchromatic, with nuclear membrane irregularities and grooves present at least focally. Frequent binucleation was observed. Small nucleoli were present in many cells, but rarely prominent. In contrast, RO showed large cells with homogenous granular cytoplasm. The nuclei showed minimal to no nuclear membrane irregularities, tiny nucleoli, mild pleomorphism, and only an occasional large, more hyperchromatic nucleus was observed. CONCLUSIONS: ChRCC has a distinct combination of cytomorphologic features. Careful attention to cytoplasmic and nuclear features allows for the distinction between ChRCC and RO in cytologic preparations. PMID- 10385449 TI - Anti-alpha-inhibin: marker of choice for the consistent distinction between adrenocortical carcinoma and renal cell carcinoma in fine-needle aspiration. AB - BACKGROUND: Anti-alpha-inhibin, an antibody directed against a peptide hormone, has been shown to be a useful diagnostic aid in surgical pathology material for the identification of sex cord-stromal neoplasms and recently has been described in adrenocortical carcinoma (ACC). The diagnosis of ACC versus renal cell carcinoma (RCC) may be difficult morphologically, particularly in fine-needle aspiration (FNA) material. To date, the immunohistochemical distinction of ACC from RCC is based on a panel of antibodies that include vimentin, cytokeratins, and epithelial membrane antigen. However, the reliability of this panel is weakened by inconsistent staining patterns. METHODS: Archival formalin fixed, paraffin embedded cell block sections from 45 FNAs of known primary and metastatic ACC and RCC as well as benign adrenocortical nodules were stained with anti-alpha-inhibin using an avidin-biotin procedure. All samples were microwave pretreated and a biotin block was performed to reduce the background stain due to the high endogenous biotin often present in these types of samples. RESULTS: All cases of ACC (n = 7; 100%) and benign adrenocortical cells (n = 15; 100%) were immunoreactive with the a-inhibin antibody, showing a diffuse cytoplasmic and granular staining pattern. The staining intensity and number of immunoreactive cells varied within each sample, with the cases of ACC having the greatest proportion of immunoreactive cells and the strongest intensity. None of the cases of RCC (n = 23; 0%) were immunoreactive with anti-alpha-inhibin. CONCLUSIONS: The morphologic distinction of ACC versus RCC in FNA material from renal, adrenal, and metastatic neoplasms is not always feasible based on cytology alone. However, due to the advent of the alpha-inhibin antibody, the reliable distinction of these entities now may be possible. The intense and specific immunostaining pattern for cells of adrenal origin, even in paucicellular samples, suggests potential for the widespread clinical utility of this marker by cytopathologists. PMID- 10385450 TI - Y2K repairs unfinished in health care sector. PMID- 10385451 TI - Amprenavir approved for HIV treatment. PMID- 10385452 TI - Are you ready for January 1, 2000? PMID- 10385453 TI - Providing timely discharge counseling. PMID- 10385454 TI - Responsible use of analgesics. PMID- 10385455 TI - Treatment of acetaminophen overdose. AB - The therapeutic management of patients with acetaminophen overdose is reviewed. Acetaminophen overdose results in more calls to poison control centers in the United States than overdose with any other pharmacologic substance. Although the optimal management strategy remains controversial, the literature suggests a general approach that can be followed until there is evidence supporting a different strategy. A single dose of activated charcoal should be administered within one hour of acetaminophen overdose. Other means of gastric decontamination are not warranted. Acetylcysteine should be given if the acetaminophen concentration exceeds the treatment line in the Rumack-Matthew nomogram. If a patient is treated within 10 hours of acetaminophen ingestion, the risk of hepatoxicity is low. In patients 10-24 hours after ingestion, a 72-hour oral or 48-hour i.v. acetylcysteine regimen should be used. Among patients with fulminant hepatic failure, acetylcysteine should be given until recovery or death occurs. In patients who have taken extended-release acetaminophen, the acetaminophen concentration should be measured at four hours and, if this level exceeds the treatment line, acetylcysteine should be started immediately. If the concentration is below the treatment line, a second acetaminophen concentration should be determined four to six hours later. If this level is above the treatment line, acetylcysteine therapy should be started. Cimetidine appears to have no role in the management of acetaminophen overdose. Children should be diagnosed and treated the same way as adults, and pregnant patients should be managed no differently than nonpregnant patients. An evaluation of the literature on acetaminophen poisoning verifies the usefulness of acetylcysteine as a hepatoprotective agent. A single dose of activated charcoal may also be useful if given within one hour of acetaminophen ingestion. PMID- 10385456 TI - Infant acceptance and effectiveness of a new oral liquid medication delivery system. AB - A medication acceptance scale (MAS) for pediatric oral liquids was developed and used to evaluate effectiveness and infant acceptance of a medication delivery system. The MAS incorporated five behavioral elements associated with pediatric drug administration: cry, facial expression, body movement or level of agitation, reaction to placement of medication in the mouth, and swallowing. A score of 1-10 was possible, with 10 indicating the highest level of infant acceptance. Preliminary field testing was conducted. In an open-label clinical study, a single dose of acetaminophen was administered to 20 infants with approximately one fluid ounce of infant formula or apple juice by pediatric nurses using the Rx medibottle (The Medicine Bottle Company). Past medication acceptance was rated on an infant global acceptance scale. The intended dose, the amount consumed, and the time taken to administer the dose were recorded. Infant acceptance was independently scored by a nurse and two pharmacists. A high preliminary estimate of internal consistency reliability of the MAS was found. Interrater reliability was high, with the highest correlation between the two pharmacists. Sixteen (80%) of the infants received 100% of the intended dose; it took 0.5-9 minutes to administer these doses. The median MAS score was 9 for each of the three raters. Mean MAS scores for the three raters were 7.85 and 7.45 (pharmacists) and 8.50 (nurse). There was a strong correlation between MAS scores and infant global acceptance scale scores. A pediatric oral liquid MAS that had content validity, concurrent validity, high internal consistency reliability, and high interrater reliability was developed; the Rx medibottle was an effective oral liquid medication delivery system and had a high level of infant and rater acceptance. PMID- 10385457 TI - Evaluating a benchmarking database and identifying cost reduction opportunities by diagnosis-related group. AB - Pharmacy cost data from the University HealthSystem Consortium (UHC) Clinical Database for specific diagnosis-related groups (DRGs) were reviewed to assess their applicability to a university medical center and to identify opportunities to reduce costs. UHC headquarters was contacted by telephone to determine UHC's data collection methods. Pharmacy costs for DRG 302 (kidney transplant) at the University of Kansas Medical Center (KUMC) were compared with the costs shown in the UHC Clinical Database. Appropriate drug use for DRGs 302 and 480 (liver transplant) was assessed by contacting transplant pharmacists and pharmacy administrators at the five top-performing hospitals (in terms of cost per DRG) as listed in the UHC database to find opportunities for reducing pharmacy costs. KUMC's actual pharmacy costs for DRG 302 ($4635) were 46% lower than those listed in the UHC Clinical Database ($8546). There was a disparity between the amount of both intravenous immune globulin (IVIG) and lymphocyte immune globulin used by KUMC and the top-performing hospitals. Guidelines for use of IVIG, acyclovir, and azathioprine in liver transplant patients at KUMC were revised. A potential cost saving of $53,000 was identified in relation to the use of lymphocyte immune globulin in kidney transplant patients. Data in the UHC Clinical Database were not representative of pharmacy costs at a university medical center for DRG 302 (kidney transplant), overstating pharmacy costs by 46%; benchmarking was found to be a useful tool for identifying opportunities for reducing costs. PMID- 10385458 TI - Rationale, design, and baseline results of the Pravastatin-to-Simvastatin Conversion Lipid Optimization Program (PSCOP). AB - A program designed to increase the percentage of patients at a Department of Veterans Affairs health system who meet their cholesterol goals as recommended by the National Cholesterol Education Program (NCEP) is described, and baseline results are reported. Patients with an active prescription for pravastatin between February 4 and June 4, 1997, were identified for conversion to simvastatin by means of the Pravastatin-to-Simvastatin Conversion Lipid Optimization Program; 1361 patients were eligible for conversion. Each patient was mailed a survey for determining risk factors for coronary heart disease (CHD) and NCEP-recommended low-density lipoprotein (LDL) cholesterol goal and was asked to provide a fasting blood sample for determination of lipid profile, liver function, and serum creatine phosphokinase concentration. The patients were asked to make a follow-up laboratory visit six to seven weeks after they had started taking simvastatin. The percentage change from baseline and the percentage of patients who meet their LDL cholesterol goal before and after the conversion will be determined. A total of 1115 patients were converted to simvastatin. Only 35.4% of patients taking pravastatin to prevent a second CHD-related event met or exceeded their LDL cholesterol goal. Only 36.2% of patients with two or more CHD risk factors who were taking pravastatin for primary prevention met or exceeded their LDL cholesterol goal. In a veterans population, less than half of patients receiving a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor had LDL cholesterol concentrations that met goals recommended by the NCEP. PMID- 10385459 TI - Prolonged neuromuscular blockade associated with rocuronium. PMID- 10385460 TI - Cancer pain management through a pharmacist-based analgesic dosing service. PMID- 10385462 TI - Pharmacy and therapeutics committee 'virtual' meetings. PMID- 10385461 TI - ASHP therapeutic position statement on the safe use of oral nonprescription analgesics. American Society of Health-System Pharmacists. AB - Nonprescription analgesics are generally safe and effective when used correctly for mild to moderate pain or fever. However, these agents do have risks, and selection of the best agent for a given patient requires consideration of a number of medication and patient factors. Consumers are encouraged to read and follow label instructions and to consult with a qualified health care provider, such as their pharmacist, for help in sorting out these factors and ensuring the best possible outcomes from nonprescription analgesic therapy. PMID- 10385463 TI - Stability of cefepime hydrochloride in polypropylene syringes. PMID- 10385465 TI - Journal references. PMID- 10385464 TI - What you need to know about...oral nonprescription pain relievers. PMID- 10385466 TI - Misquoting and quoting out of context. PMID- 10385467 TI - Certainty, leaps of faith and tradition: rethinking clinical interventions. PMID- 10385468 TI - Sailing beyond: nursing theory and the person. AB - This article explores nursing's conceptualization of the person in terms of holism and uniqueness. These concepts raise concerns from postmodernist and feminist perspectives in terms of the unitary self, essentialism, culturalism, agency, and subjectivity. The use of holism and uniqueness in nursing theory appeals to the generic person, disregards interactions among the individual's race, class, and sex, and, thus, leaves larger institutional and societal issues unacknowledged, unexplored, and unchallenged. Nursing theories are needed that have social justice as their goal. PMID- 10385469 TI - Caring and the science of unitary human beings. AB - The purpose of this article is to clarify the ambiguity surrounding the concept of caring through situating it within one conceptual system, the Science of Unitary Human Beings. An analysis of the dialogue on caring in nursing is presented. A process of concept clarification was developed to examine points of congruence between existing literature on caring and theoretical niches expressing similar meanings in the Science of Unitary Human Beings. The process resulted in the synthesis of five constitutive meanings of caring in the Science of Unitary Human Beings: manifesting intentions, appreciating pattern, attuning to dynamic flow, experiencing the infinite, and inviting creative emergence. Narratives were developed to ground the abstract meanings in concrete human experience. PMID- 10385470 TI - The ecologies of community caring. AB - Caring has been called the interactive exemplar of nursing in that it relates other foundational concepts of person, health, and environment within the profession and discipline. However, to reflect the dynamic praxis of knowing, being, and doing that is community health nursing, caring must include a focus on communities, the environment, and the global society. The concepts of community and community interventions remain complex and difficult, thus, community caring remains unclear. This article will explore the concept and offer a model of caring praxis in community health nursing. PMID- 10385471 TI - Critical nursing scholarship: exploring critical social theory with African American studies. AB - There is a need for nursing research that applies ideas gained from critical nursing scholarship, yet attends to the historical, cultural, and social context of clients analyzed through those frameworks. The revision of critical nursing scholarship to address multicultural perspectives on critical thought will significantly transform nursing knowledge development. This article first provides overviews about critical nursing scholarship and critical social theory. It then explores the union of critical social theory and African American studies. Ideas gained from the combination of critical social theory and African American studies are incorporated subsequently in discussions about the role of theory in representations of social identity, in the development of knowledge, and in the development of nursing research. PMID- 10385472 TI - Toward an understanding of art in nursing. AB - The art of nursing is a term on which there is little definitional agreement. Nursing includes both artistic and scientific traditions; at times, the balance between these two approaches is an uneasy one. An examination of aesthetics offers a perspective to ground practice in nursing art through John Dewey's philosophy of art as experience. Differences and similarities between science, art, and craft are examined. Nursing art is defined as helping patients create coherence in lives threatened by illness and change. PMID- 10385473 TI - Multiple paradigms of nursing science. AB - Of the many controversies that have accompanied the growth of nursing as a discipline, few have been debated as long and as vigorously as the question of which paradigm should guide nursing science. Despite more than 20 years of discussion, the question remains unresolved. This article discusses the concept of paradigm, explores the paradigms that influence nursing science, and presents a comparison of the advantages and disadvantages of theoretical unification and mutiparadigmism. Additionally, the implications and consequences of multiparadigmism for the present and future development of nursing as a science within a practice discipline are presented. PMID- 10385474 TI - Middle range theory: spinning research and practice to create knowledge for the new millennium. AB - The foundation of middle range theory reported during the past decade was described and analyzed. A CINAHL search revealed 22 middle range theories that met selected criteria. This foundation is a firm base for new millennium theorizing. Recommendations for future theorizing include: clear articulation of theory names and approaches for generating theories; clarification of concept linkages with inclusion of diagrammed models; deliberate attention to research practice connections of theories; creation of theories in concert with the disciplinary perspective; and, movement of middle range theories to the front lines of nursing research and practice for further analysis, critique, and development. PMID- 10385475 TI - It's as simple as black and white! Race and ethnicity as categorical variables. PMID- 10385476 TI - Chemokines and renal inflammation in proteinuric disorders: searching for the inciting stimulus. PMID- 10385477 TI - Benign ethnic neutropenia: what is a normal absolute neutrophil count? AB - Approximately 25% to 50% of persons of African descent and some ethnic groups in the Middle East have benign ethnic neutropenia, with low leukocyte and neutrophil counts. It is important to recognize the existence of this condition, the most common form of neutropenia throughout the world, and thus avoid both under-and overevaluation. Although there is no scientific basis for an absolute neutrophil count of 1.5x10(9)/L to be considered minimal, counts below this level are empirically regarded as inadequate in persons of all ethnic groups who are above the age of 1 year. Many individuals, however, maintain consistently low absolute neutrophil counts without evidence of increased susceptibility to infection or any other adverse effect. The important determination is not how many neutrophils are present in the peripheral blood, but whether the bone marrow is able to produce enough normally functioning cells when needed. A description of benign ethnic neutropenia, as set forth in this review, suggests that the lower limit now considered acceptable for the absolute neutrophil count should be readjusted downward for all ethnic groups. PMID- 10385478 TI - Genomic imprinting of the X chromosome: a novel mechanism for the evolution of sexual dimorphism. AB - Genomic imprinting is the differential marking of maternally and paternally inherited alleles of specific genes or chromosome regions during gametogenesis. The imprint silences the allele from 1 parent. A number of imprinted genes that are expressed in the brain have been identified in humans. They control the actions of other genes or regulate their products. Sexual dimorphism in the vertebrate brain is conventionally thought to be due to the epigenetic action of gonadal hormones. Sex differences could also reflect the actions of an imprinted X-linked locus. Until very recently no imprinted gene had been described on the X chromosome in humans. Here the implications of such a mechanism for the evolution of sexual dimorphism are discussed. PMID- 10385479 TI - Ankle brachial index as a predictor of outcomes in peripheral arterial disease. PMID- 10385480 TI - Gene expression of CC chemokines in experimental acute tubulointerstitial nephritis. AB - The infiltration of mononuclear leukocytes into glomeruli or the interstitium is a feature in most forms of glomerular diseases. CC chemokines, mostly chemoattractants for mononuclear leukocytes, are molecules that are potentially responsible for the recruitment of these cells in the kidney. We previously reported that the gene expression of six CC chemokines-MCP-1, MCP-3, MIP-1alpha, MIP-1beta, RANTES, and TCA3-was enhanced in a rat model of crescentic glomerulonephritis, the most severe form of glomerulonephritis. In this study we analyzed their gene expression in a model of another type of kidney disease, acute nephrosis accompanied by tubulointerstitial lesions, which is induced by an injection of puromycin aminonucleoside. Because leukocyte infiltration in this model is much more prominent in the interstitium than in glomeruli, we analyzed their gene expression in the renal cortex. On day 3, when the level of urinary protein was slightly but significantly increased but the number of interstitial leukocytes was unchanged, the enhanced expression of mRNAs for MCP-1, MCP-3, and TCA3 was observed. On day 5, the numbers of interstitial monocytes and lymphocytes significantly increased, and the levels of the mRNA expression of the above chemokines were still higher than the control animals, whereas the levels of mRNAs for MIP- 1alpha, MIP-1beta, and RANTES were not higher or were only slightly higher than the control ones. These results suggest that multiple CC chemokines may play a role in the recruitment of leukocytes in this model and that the expression pattern of CC chemokines depends on the type of kidney injury. PMID- 10385481 TI - Acute infusion of nicotine potentiates norepinephrine-induced vasoconstriction in the hamster cheek pouch. AB - Although cigarette smoking and the components of cigarette smoke appear to alter nitric oxide synthase-dependent dilation of blood vessels, the effect of these substances on constrictor responses of resistance arterioles has not been examined. Thus the goal of this study was to examine the effect of a major component of cigarette smoke-that is, nicotine-on constrictor responses of cheek pouch arterioles. The diameter of cheek pouch arterioles (approximately 50 microm in diameter) was measured by using intravital microscopy. We examined the responses of arterioles to angiotensin II, arginine vasopressin, norepinephrine, and the thromboxane analog U-46619 before and after treatment with vehicle (saline solution), N(G)-monomethyl-L-arginine (L-NMMA; 1.0 micromol/L), or nicotine (2.0 microg/kg/min i.v. for 30 minutes followed by a maintenance dose of 0.35 microg/kg/min for 30 minutes). Topical application of angiotensin II (0.01 and 0.1 nmol/L), arginine vasopressin (1.0 and 10 pmol/L), norepinephrine (1.0 and 10 nmol/L), and U-46619 (0.01 and 0.1 nmol/L) produced marked reproducible constriction of cheek pouch arterioles in hamsters treated with vehicle. Topical application of L-NMMA potentiated constrictor responses of arterioles to the high dose of arginine vasopressin (28%+/-4% versus 36%+/-4%; P<.05) and to both doses of norepinephrine (14%+/-1% and 24%+/-2% versus 19%+/-1% and 31%+/-3%; P<.05). The infusion of nicotine did not alter responses to angiotensin II, arginine vasopressin, or U-46619 but modestly potentiated vasoconstriction in response to norepinephrine (12%+/-2% and 22%+/-2% versus 14%+/-2% and 26%+/-2%; P<.05). These findings suggest that the synthesis/release of nitric oxide may modulate constrictor responses of cheek pouch resistance arterioles to selected agonists. In addition, nicotine, at levels observed in smokers, may potentiate norepinephrine-induced vasoconstriction. We suggest that preservation/potentiation of vasoconstrictor responses may contribute to the pathogenesis of vascular abnormalities associated with cigarette smoking. PMID- 10385482 TI - Inhibition of activated human mesangial cell proliferation by the natural product of Cordyceps sinensis (H1-A): an implication for treatment of IgA mesangial nephropathy. AB - Cordyceps sinensis (CS) is a parasitic fungus that has been used as a Chinese medicine for a long time in the treatment of nephritis. Today, the hypothesis about the pathogenesis of immunoglobulin A nephropathy (IgAN) is that nephritogenic IgA immune complexes (IgAIC) go to the kidney to stimulate resting mesangial cells to release cytokines and growth factors. These cytokines and growth factors cause mesangial cell proliferation and release matrix, chemical mediators that lead to the glomerular injury. However, nephritogenic IgAIC in humans is still unknown. To solve this problem previously, we established an in vitro model that showed that cultured human mesangial cells (HMC) stimulated with interleukin-1 (IL-1) plus IL-6 can cause mesangial cell proliferation, increasing production of chemical mediators and superoxide anion. An in vivo model also proved that this culture medium may lead to renal injury with hematuria and proteinuria. Therefore, to fractionate the crude components that can be used in the treatment of patients with IgAN, we cultured HMC, and then an HMC activating model with HMC incubated with IL-1 and IL-6 was established. We fractionated the crude methanolic extracts from fruiting bodies of CS with the use of this in vitro inhibition of HMC activation model as our assay method. In brief, the fruiting bodies were extracted by silica gel column chromatography. One out of 6 column fractions, F-2, significantly inhibited the HMC activation by IL-1 plus IL 6. The acute toxicity test with male Institute of Cancer Research mice showed no liver toxicity or mutagenicity. Then we established an IgAN animal model with R36A (Pneumococcal C-polysaccharide purified from Streptococcus pneumoniae) as antigen and anti-R36A IgA monoclonal antibody to form nephritogenic IgA-IC, which can induce hematuria and proteinuria in mice with IgA deposition in the mesangial area. The mice in the IgAN model fed with 1% F-2 in diet had significant reduction of hematuria and proteinuria together with histopathologic improvement. Therefore this fraction was then purified by silica gel column chromatography and high-performance liquid chromatography, which got a purified compound H1-A, which can suppress the activated HMC and alleviate IgAN (Berger's disease) with clinical and histologic improvement. These results give us a new regimen for the treatment of patients with IgAN in the future. PMID- 10385484 TI - Impaired metoclopramide-induced pituitary prolactin release in men with human immunodeficiency virus infection. AB - To investigate whether metoclopramide-induced prolactin release is impaired in HIV-infected men, we studied 10 clinically healthy HIV-negative adult men (group 1) and 10 consecutive HIV-positive adult men (group 2) with anti-HIV antibodies confirmed by Western blot analysis and a CD4 cell count from 13 to 570x10(6)/L. After a 10- to 12-hour overnight fast, three basal blood samples were obtained at 15-minute intervals (-30, -15, and 0 minutes) and thereafter at 15, 30, 60, 90, 120, 180, and 240 minutes after a 10-mg intravenous bolus of metoclopramide. Duplicate serum prolactin concentrations were measured in each sample with commercially available radioimmunoassay kits. No significant differences between groups were observed in basal prolactin levels. Group 2 had lower serum prolactin concentrations than group 1 throughout the test (P< or =.002). The area under the prolactin curve (mean +/- SD) was also lower in group 2 than in group 1 (7156+/ 1433 ng/mL/240 min vs. 12430+/-2454 ng/mL/240 min, P<.0001), and the area under the prolactin curve had a significant correlation with the CD4 cell counts (r = 0.7912, P<.001). These findings suggest that the hypothalamic dopaminergic tone, although present, was clearly diminished in these HIV-positive men regardless of their clinical stage. PMID- 10385483 TI - Deferiprone therapy in homozygous human beta-thalassemia removes erythrocyte membrane free iron and reduces KCl cotransport activity. AB - Deposition of free iron is a characteristic feature of beta-thalassemia (beta thal) red blood cells believed to play an important role in the generation of oxidative injury to the cell membrane. Increased red blood cell KCI cotransport, reduced K content, and cell dehydration are also found in beta-thal red blood cells. It is not known, however, whether deposition of free iron plays a role in these membrane transport changes. To explore this issue, we studied-both in vitro and in vivo-the effect on KCI cotransport of removing red blood cell membrane free iron from beta-thal erythrocytes. Eleven patients with beta-thal major who underwent long-term transfusion and were treated with deferiprone (75 mg/kg/day) for 9 months participated in the study. Deferiprone therapy removed membrane free iron from beta-thal erythrocytes, which was followed by reduced KCI cotransport activity. The reduced KCI cotransport activity was accompanied by an increase in the red blood cell K content. These data suggest that the increased activity of KCI cotransport in beta-thal red blood cells is mediated by the deposition of membrane free iron, a mechanism that may be attenuated by deferiprone therapy. PMID- 10385485 TI - Plasma leptin concentrations in lean and obese human subjects and Prader-Willi syndrome: comparison of RIA and ELISA methods. AB - Immunoassays for circulating leptin are important research tools for examining the role and regulation of leptin expression in human obesity. However, uncertainty exists regarding the comparability between studies of reported plasma or serum leptin concentrations. The purpose of the present study was to directly compare plasma leptin concentrations by using two of the most widely reported immunoassay methods-namely, a commercially available radioimmunoassay (RIA) and a proprietary enzyme-linked immunosorbent assay (ELISA). Plasma leptin concentrations were measured in healthy lean and obese volunteers and in patients with Prader-Willi syndrome (PWS). Over a wide range of plasma concentrations (2 to 70 ng/mL), leptin measurements obtained with the RIA and ELISA methods were highly correlated (r = 0.957, P<.0001) and were essentially indistinguishable. Leptin levels measured by RIA and ELISA were highly correlated with body mass index (BMI) overall (r = 0.784, P<.0001 and r = 0.732, P<.0001, respectively) and in the lean and obese subgroups. When compared with the results in the lean individuals (mean +/- SEM, 11.6+/-3.2 ng/mL), plasma leptin was significantly higher in both the obese (35.5+/-4.0 ng/mL, P<.0001) and the PWS subjects (30.7+/ 6.9 ng/mL, P<.05). However, after we controlled for differences in BMI, the leptin levels were similar in all three groups. In conclusion, we found that the RIA and ELISA used in the present study yield plasma leptin concentrations that are essentially indistinguishable. Our findings should facilitate comparisons of leptin levels measured by these two widely used immunoassays in previous and future studies that examine the role of leptin in body weight regulation. PMID- 10385486 TI - Uptake of heparin cofactor II and antithrombin into the aorta wall after a deendothelializing injury in vivo: comparison with the behaviors of prothrombin and fibrinogen. AB - The initiation of a denuding injury to the vascular endothelium rapidly leads to a deposition of platelets and fibrin at the site of injury. We have measured previously the responses of rabbit fibrinogen, prothrombin, and antithrombin to a deendothelializing balloon-catheter injury to the rabbit aorta in vivo. In this study, rabbit iodine 125-labeled HCII and iodine 125-labeled AT were coinjected intravenously into anesthetized rabbits 5 minutes before deendothelialization of the thoracic aorta. The rabbit was exsanguinated at 5 to 60 minutes after injury, the aorta was excised, and the accumulation of each radiolabeled protein in each layer of aorta wall was determined relative to the concentration of the respective native protein in circulating blood at exsanguination. The maximum flux rates into the aorta wall (i.e., platelet layer and intima-media) in the first minute after injury were calculated from the uptake data; approximately 2.8 molecules of AT accumulated for each HCII molecule. By comparison with previous measurements, the maximum flux rate of AT was similar to that of prothrombin. Further, the molar ratio of accumulated prothrombin/AT + HCII) in the aorta wall was 0.75. Detergent extracts of the injured aorta intima-media contained unreacted HCII and HCII complexes; the uninjured aorta contained only unreacted HCII. By contrast, high molecular weight AT complexes and unreacted AT were extracted from the uninjured, and in greater quantity from the injured, aorta wall. We conclude that, of the plasma antithrombins, AT accumulated more rapidly than HCII in vivo and appeared to be the more active inhibitor at the site of vascular injury. HCII may play a relatively minor role as an antithrombin and possibly only after injury. PMID- 10385488 TI - Circulation online only : june 29, 1999 PMID- 10385489 TI - Polymorphism of the lipopolysaccharide receptor (CD14) and myocardial infarction. New evidence for a role of gram-negative bacterial infection? PMID- 10385490 TI - TNF-alpha and heart failure. The difference between proof of principle and hypothesis testing. PMID- 10385491 TI - Cholesterol lowering. Should it continue to be the last thing we do? PMID- 10385492 TI - C(-260)-->T polymorphism in the promoter of the CD14 monocyte receptor gene as a risk factor for myocardial infarction. AB - BACKGROUND: The CD14 receptor of monocytes is an important mediator for the activation of monocytes/macrophages by endotoxins from the envelope of Gram negative bacteria (lipopolysaccharides). We identified a polymorphism in the CD14 receptor and examined whether this genetic marker influenced the expression of the CD14 receptor on monocytes and affected the predisposition to myocardial infarction. METHODS AND RESULTS: We identified a C(-260)-->T nucleotide change, creating a HaeIII polymorphism in the promoter of the CD14 gene. The polymorphism was determined in 178 male patients <65 years old (cases; average age, 55.9+/-6.3 years) at the time of their first myocardial infarction and in 135 representative selected male control subjects (controls; average age, 55.2+/-11.5 years). The frequency of the T allele (absence of the cutting site) was 0.49 in cases and 0.35 in controls (P=0.0005; OR, 1.781; 95% CI, 1.286 to 2.465). Subsequently, we measured the expression of monocyte CD14 by flow cytometry in 18 volunteers with different CD14 genotypes. A significantly higher density of the CD14 receptor was shown in the T/T homozygotes than in the others (P=0.0028). CONCLUSIONS: A higher frequency of allele T(-260) in the promoter of the CD14 receptor gene was found in myocardial infarction survivors than in controls. At the same time, this variation was associated with a higher density of CD14 receptors in healthy volunteers. Therefore, we can conclude that in addition to the well-established risk factors, a genetically determined reaction of monocytes/macrophages to infectious stimuli could play an important role in the process of atherosclerosis. PMID- 10385493 TI - Prevention of distal embolization during saphenous vein graft lesion angioplasty. Experience with a new temporary occlusion and aspiration system. AB - BACKGROUND: Repeat coronary artery bypass graft surgery (CABG) is associated with a high morbidity and mortality, rendering percutaneous treatment of saphenous vein graft (SVG) lesions an attractive alternative. However, percutaneous interventions of degenerated SVGs carries high risk of distal embolization. METHODS AND RESULTS: This study reports our initial experience with the PercuSurge GuardWire, a new device developed to prevent embolization during treatment of degenerated SVG. This device consists of a 190-cm-long, hollow 0.014 in guidewire with a central lumen connected to a distal occlusion balloon. A dedicated inflation device (the MicroSeal Adapter) was used to inflate the distal balloon and maintain complete lumen occlusion during balloon dilatation and stent implantation. A monorail aspiration catheter, connected to a vacuum syringe, was used to evacuate atherosclerotic and thrombotic debris. Angioplasty with stent implantation was performed in 15 degenerated SVGs (18 lesions). Procedural success was achieved in all patients with normal postprocedure flow (Thrombolysis in Myocardial Infarction grade 3). No distal embolization was observed. There were no major in-hospital adverse clinical events, including Q-wave or non-Q-wave myocardial infarction, emergency CABG, or death. All patients were asymptomatic at discharge. CONCLUSIONS: This preliminary series supports the feasible use of the PercuSurge GuardWire for retrieval of plaque debris and prevention of embolization in degenerated SVGs. The good tolerance of temporary occlusions without angiographic or clinical evidence of distal embolization represents encouraging early findings. PMID- 10385494 TI - Safety and efficacy of a soluble P75 tumor necrosis factor receptor (Enbrel, etanercept) in patients with advanced heart failure. AB - BACKGROUND: Although previous studies suggested that TNF may contribute to heart failure progression, it is unclear whether antagonizing TNF is beneficial in heart failure patients. METHODS AND RESULTS: Eighteen NYHA class III heart failure patients were randomized into a double-blind dose-escalation study to examine the safety and potential efficacy of etanercept, a specific TNF antagonist (Enbrel). Patients received placebo (6 patients) or an escalating dose (1, 4, or 10 mg/m2) of etanercept (12 patients) given as a single intravenous infusion. Safety parameters and patient functional status were assessed at baseline and at days 1, 2, 7, and 14. There were no significant side effects or clinically significant changes in laboratory indices. There was, however, a decrease in TNF bioactivity and a significant overall increase in quality-of-life scores, 6-minute walk distance, and ejection fraction in the cohort that received 4 or 10 mg/m2 of etanercept; there was no significant change in these parameters in the placebo group. CONCLUSIONS: A single intravenous infusion of etanercept was safe and well tolerated in patients with NYHA class III heart failure. These studies provide provisional evidence that suggests that etanercept is sufficient to lower levels of biologically active TNF and may lead to improvement in the functional status of patients with heart failure. PMID- 10385495 TI - Cholesterol reduction rapidly improves endothelial function after acute coronary syndromes. The RECIFE (reduction of cholesterol in ischemia and function of the endothelium) trial. AB - BACKGROUND: Cholesterol lowering reduces coronary events. One mechanism could be improvement of endothelial function. In line with this hypothesis, this study investigates whether cholesterol reduction can result in rapid improvement of endothelial function after acute coronary syndromes. METHODS AND RESULTS: Patients with acute myocardial infarction or unstable angina and total cholesterol levels at admission >/=5.2 mmol/L or LDL >/=3.4 mmol/L were randomized to placebo (n=30) or pravastatin 40 mg daily (n=30) for 6 weeks. Brachial ultrasound was used to measure endothelium-dependent flow-mediated dilatation (FMD) and response to endothelium-independent nitroglycerin. Changes in the levels of markers of platelet activation, coagulation factors, and plasma endothelin levels were also assessed. Total and LDL cholesterol levels were similar at admission and before randomization in both groups. With pravastatin, but not with placebo, they decreased by 23% (P<0.05) and 33% (P<0.01), respectively. FMD was unchanged with placebo, 5.43+/-0.74% (mean+/-SEM) to 5.84+/ 0.81%, but increased with pravastatin, 4.93+/-0.81% to 7.0+/-0.79% (P=0.02), representing a 42% relative increase. Responses to nitroglycerin were similar during the time course of the study in the 2 groups. Markers of platelet activity, coagulation factors, and endothelin levels were not affected by pravastatin. CONCLUSIONS: Cholesterol reduction with pravastatin initiated early after acute coronary syndromes rapidly improves endothelial function after 6 weeks of therapy. PMID- 10385496 TI - Long-term ascorbic acid administration reverses endothelial vasomotor dysfunction in patients with coronary artery disease. AB - BACKGROUND: Loss of endothelium-derived nitric oxide (EDNO) contributes to the clinical expression of coronary artery disease (CAD). Increased oxidative stress has been linked to impaired endothelial vasomotor function in atherosclerosis, and recent studies demonstrated that short-term ascorbic acid treatment improves endothelial function. METHODS AND RESULTS: In a randomized, double-blind, placebo controlled study, we examined the effects of single-dose (2 g PO) and long-term (500 mg/d) ascorbic acid treatment on EDNO-dependent flow-mediated dilation of the brachial artery in patients with angiographically established CAD. Flow mediated dilation was examined by high-resolution vascular ultrasound at baseline, 2 hours after the single dose, and 30 days after long-term treatment in 46 patients with CAD. Flow-mediated dilation improved from 6.6+/-3.5% to 10.1+/ 5.2% after single-dose treatment, and the effect was sustained after long-term treatment (9. 0+/-3.7%), whereas flow-mediated dilation was 8.6+/-4.7% at baseline and remained unchanged after single-dose (7.8+/-4.4%) and long-term (7.9+/-4.5%) treatment with placebo (P=0.005 by repeated-measures ANOVA). Plasma ascorbic acid concentrations increased from 41.4+/-12. 9 to 115.9+/-34.2 micromol/L after single-dose treatment and to 95. 0+/-36.1 micromol/L after long term treatment (P<0.001). CONCLUSIONS: In patients with CAD, long-term ascorbic acid treatment has a sustained beneficial effect on EDNO action. Because endothelial dysfunction may contribute to the pathogenesis of cardiovascular events, this study indicates that ascorbic acid treatment may benefit patients with CAD. PMID- 10385497 TI - Aggressive cholesterol lowering delays saphenous vein graft atherosclerosis in women, the elderly, and patients with associated risk factors. NHLBI post coronary artery bypass graft clinical trial. Post CABG Trial Investigators. AB - BACKGROUND: The NHLBI Post Coronary Artery Bypass Graft trial (Post CABG) showed that aggressive compared with moderate lowering of low-density lipoprotein cholesterol (LDL-C) decreased obstructive changes in saphenous vein grafts (SVGs) by 31%.1 Using lovastatin and cholestyramine when necessary, the annually determined mean LDL-C level ranged from 93 to 97 mg/dL in aggressively treated patients and from 132 to 136 mg/dL in the others (P<0.001). METHODS AND RESULTS: The present study evaluated the treatment effect in subgroups defined by age, gender, and selected coronary heart disease (CHD) risk factors, ie, smoking, hypertension, diabetes mellitus, high-density lipoprotein cholesterol (HDL-C) <35 mg/dL, and triglyceride serum levels >/=200 mg/dL at baseline. As evidenced by similar odds ratio estimates of progression (lumen diameter decrease >/=0.6 mm) and lack of interactions with treatment, a similar beneficial effect of aggressive lowering was observed in elderly and young patients, in women and men, in patients with and without smoking, hypertension, or diabetes mellitus, and those with and without borderline high-risk triglyceride serum levels. The change in minimum lumen diameter was in the same direction for all subgroup categories, without significant interactions with treatment. CONCLUSIONS: Aggressive LDL-C lowering delays progression of atherosclerosis in SVGs irrespective of gender, age, and certain risk factors for CHD. PMID- 10385498 TI - Visualization and functional assessment of proximal and middle left anterior descending coronary stenoses in humans with magnetic resonance imaging. AB - BACKGROUND: Coronary artery bypass grafting improves survival in patients with >70% luminal diameter narrowing of the 3 major epicardial coronary arteries, particularly if there is involvement of the proximal portion of the left anterior descending (LAD) coronary artery. Measurement of coronary flow reserve can be used to identify functionally important luminal narrowing of the LAD artery. Although magnetic resonance imaging (MRI) has been used to visualize coronary arteries and to measure flow reserve noninvasively, the utility of MRI for detecting significant LAD stenoses is unknown. METHODS AND RESULTS: Thirty subjects (23 men, 7 women, age 36 to 77 years) underwent MRI visualization of the left main and LAD coronary arteries as well as measurement of flow in the proximal, middle, or distal LAD both at rest and after intravenous adenosine (140 microgram/kg per minute). Immediately thereafter, contrast coronary angiography and when feasible, intracoronary Doppler assessments of coronary flow reserve, were performed. There was a statistically significant correlation between MRI assessments of coronary flow reserve and (a) assessments of coronary arterial stenosis severity by quantitative coronary angiography and (b) invasive measurements of coronary flow reserve (P<0.0001 for both). In comparison to computer-assisted quantitative coronary angiography, the sensitivity and specificity of MRI for identifying a stenosis >70% in the distal left main or proximal/middle LAD arteries was 100% and 83%, respectively. CONCLUSIONS: Noninvasive MRI measures of coronary flow reserve correlated well with similar measures obtained with the use of intracoronary Doppler flow wires and predicted significant coronary stenoses (>70%) with a high degree of sensitivity and specificity. MRI-based measurement of coronary flow reserve may prove useful for identification of patients likely to obtain a survival benefit from coronary artery bypass grafting. PMID- 10385499 TI - Five-year outcome in patients with isolated proximal left anterior descending coronary artery stenosis treated by angioplasty or left internal mammary artery grafting. A prospective trial. AB - BACKGROUND: Percutaneous transluminal coronary angioplasty (PTCA) and coronary artery bypass surgery (CABG) improve the clinical status of patients with isolated proximal left anterior descending coronary artery stenosis. At 2 years, only additional revascularization was more frequently required after PTCA. METHODS AND RESULTS: We monitored 134 patients randomized to PTCA (n=68) or CABG (n=66) for /=45 mm) showed significantly greater expression of TACE (P=0.02) and TNF-alpha (P=0. 001) than did the low LVESD subgroup (<45 mm). In addition, the DCM subgroup with lower LVEF (<40%) showed higher expression of TACE (P=0.006) and TNF-alpha (P=0.01) than did the subgroup with high LVEF (>/=40%). CONCLUSIONS: This study has shown that increased myocardial TACE expression is associated with elevated myocardial TNF alpha expression in both mRNA and protein levels in clinically advanced DCM. PMID- 10385501 TI - Continuous infusion of epoprostenol improves the net balance between pulmonary endothelin-1 clearance and release in primary pulmonary hypertension. AB - BACKGROUND: Primary pulmonary hypertension results from progressive narrowing of the precapillary pulmonary vasculature. A variety of endothelial abnormalities have been identified, including a net reduction in pulmonary clearance of the vasoconstrictor and smooth muscle mitogen endothelin-1. In many patients, net pulmonary release of endothelin-1 is observed. Chronic infusions of epoprostenol (prostacyclin) improve functional capacity, survival, and hemodynamics in patients with advanced primary pulmonary hypertension. We hypothesized that the epoprostenol infusions, as compared with conventional therapy, might alter the abnormal pulmonary endothelin-1 homeostasis. METHODS AND RESULTS: Using a subset of patients from a larger randomized study comparing epoprostenol plus conventional therapy (n=11 in the present study) with conventional therapy alone (n=7 in the present study), we determined the ratio of plasma endothelin-1 levels in systemic arterial blood leaving the lung to levels in mixed venous blood entering the lung both before randomization and after 88 days of continuous therapy. There were no differences between the 2 groups before therapy, but by day 88, the epoprostenol-treated group had a greater proportion of patients (82%) with an arterial/venous ratio <1 than did the conventional therapy group, in which only 29% of patients had a ratio <1 (P<0.05). CONCLUSIONS: These results suggest that continuous epoprostenol therapy may have a beneficial effect on the balance between endothelin-1 clearance and release in many patients with primary pulmonary hypertension and may provide one explanation for the salutary effect of epoprostenol in this disease. PMID- 10385502 TI - Late results of percutaneous mitral commissurotomy in a series of 1024 patients. Analysis of late clinical deterioration: frequency, anatomic findings, and predictive factors. AB - BACKGROUND: The optimal use of percutaneous mitral commissurotomy (PMC) in a wide range of patients requires accurate evaluation of late results and identification of their predictors. METHODS AND RESULTS: Late results of PMC were assessed in 1024 patients whose mean age was 49+/-14 years. Echocardiography showed that 141 patients (14%) had pliable valves and mild subvalvular disease, 569 (55%) had extensive subvalvular disease, and 314 (31%) had calcified valves. A single balloon was used in 26 patients, a double balloon in 390, and the Inoue Balloon in 608. Good immediate results were defined as valve area >/=1.5 cm2 without regurgitation >2/4 (Sellers' grade) and were obtained in 912 patients. Median duration of follow-up was 49 months. The 10-year actuarial rate of good functional results (survival with no cardiovascular death and no need for surgery or repeat dilatation and in New York Heart Association [NYHA] class I or II) was 56+/-4% in the entire population. Follow-up echocardiography was available in 90% of the patients who experienced poor functional results after good immediate results and showed restenosis in 97% of these. In multivariate analysis, the predictors of poor functional results were old age (P=0.0008), unfavorable valve anatomy (P=0.003), high NYHA class (P<0.0001), atrial fibrillation (P<0.0001), low valve area after PMC (P=0.001), high gradient after PMC (P<0.0001), and grade 2 mitral regurgitation after PMC (P=0.04). CONCLUSIONS: PMC can be performed with good late results in a variety of patient subsets. Prediction of late events is multifactorial. Knowledge of these predictors can improve patient selection and follow-up. PMID- 10385503 TI - Origins of heart rate variability. Inducibility and prevalence of a discrete, tachycardic event. AB - BACKGROUND: We propose that heart period sequences are linearly organized, like sentences, and that there is a lexicon of recurrent, similarly shaped transient structures like words. Each word (or lexon) has a characteristic physiological basis. One potential lexon is the transient, reversible tachycardia that is induced by exercise initiation under laboratory conditions. We hypothesized that this lexon was inducible and observable on ambulatory ECGs of most or all subjects, was morphologically similar in both induced and detected bursts, and shared a plausible origin in both circumstances. METHODS AND RESULTS: Ten healthy subjects (mean age, 36 years) underwent a protocol in which subjects rolled themselves from supine to lateral decubitus positions and back. Transient tachycardias ("bursts") were seen in 36 of 40 rollovers. Bursts were characterized by an initial monoexponential heart period decay (K=0.39+/-0.23 s 1), a maximum heart period decrease of 277+/-109 ms after 10.8+/-4.5 seconds, and a subsequent return to baseline 23.3+/-10.8 seconds after roll initiation. The roll-induced bursts were detected with 97% sensitivity and 99% specificity with a search algorithm that incorporated morphological parameters. In 24-hour ambulatory ECGs of 10 healthy subjects (mean age, 38 years; range, 17 to 69 years), 117+/-59 bursts were detected. Induced and detected bursts were similar in most morphological parameters. Finally, many bursts occurred at night, when rolling over also occurs. CONCLUSIONS: Bursts are inducible, transient tachycardias that occur clinically and constitute a lexon with an understandable physiology. PMID- 10385504 TI - Isoproterenol to evaluate resumption of conduction after right atrial isthmus ablation in type I atrial flutter. AB - BACKGROUND: After radiofrequency (RF) ablation of atrial flutter (AFL), the demonstration of bidirectional isthmus conduction (BIC) block is considered the hallmark of a successful procedure. The purpose of our study was to test the persistence of BIC block after isoproterenol administration and to evaluate the importance of this finding with regard to AFL recurrences. METHODS AND RESULTS: RF ablation of AFL was performed in 44 consecutive patients with type I AFL by linear ablation of the posterior isthmus (n=29 patients), septal isthmus (n=4 patients), or both right atrial (RA) isthmi (n=11 patients). The procedural end point was complete BIC block and noninducibility of AFL. In case of noninducibility and apparent BIC block, the pacing protocol was repeated under isoproterenol infusion (1 to 3 microgram/min). Reversal of apparent BIC block occurred in 7 (15.9%) of 44 patients. Six patients had bidirectional and 1 had unidirectional resumption of isthmus conduction. Counterclockwise AFL could be reinduced in 4 of these patients. Two to 24 (median, 4) additional RF applications were required to achieve permanent BIC block. At a mean follow-up of 7.3+/-7.6 months (range, 2 to 31 months), 2 (4.5%) of 44 patients had AFL recurrences. CONCLUSIONS: Partial linear RF ablation could possibly aggravate preexisting nonuniform anisotropic conduction in the RA isthmus, resulting in profound conduction slowing and apparent BIC block. Isoproterenol can unmask apparent BIC block, thus providing an opportunity to assess the possibility of reversal of BIC block and completeness of isthmus ablation during the same procedure. The low incidence (4.5%) of AFL recurrences at follow-up suggests that noninducibility and BIC block under isoproterenol infusion may be a better end point for successful AFL ablation. PMID- 10385505 TI - Restenosis following angioplasty in the swine coronary artery is inhibited by an orally active PDGF-receptor tyrosine kinase inhibitor, RPR101511A. AB - BACKGROUND: Platelet-derived growth factor (PDGF), a purported mediator of arterial response to injury, stimulates proliferation, chemotaxis, and matrix production by activation of its membrane receptor tyrosine kinase. Because these activities underlie restenosis, inhibition of the PDGF-receptor tyrosine kinase (PDGFr-TK) is postulated to decrease restenosis. METHODS AND RESULTS: RPR101511A is a novel compound which selectively and potently inhibits the cell-free and in situ PDGFr-TK and PDGFr-dependent proliferation and chemotaxis in vascular smooth muscle cells (VSMC). To evaluate the effect of RPR101511A (30 mg. kg-1. d-1 BID for 28 days following PTCA) on coronary restenosis, PTCA was performed in hypercholesterolemic minipigs whose left anterior descending (LAD) coronary artery had been injured by overdilation and denudation, yielding a previously existing lesion. Angiographically determined prePTCA minimal lumen diameters (MLD) were similar in vehicle and RPR101511A-treated pigs (1.98+/-0.09 versus 2.01+/-0.08 mm) and increased to the same extent in the 2 groups following successful PTCA (2.30+/-0.06 versus 2.52+/-0.13). At termination, there was an average 50% loss of gain in the vehicle-treated group but no loss of gain with RPR101511A (2.16+/-0. 05 versus 2.59+/-0.11, P<0.001). Morphometric analysis of the LAD showed that RPR101511A caused a significant decrease in total intimal/medial ratio (0.96+/-0.58 versus 0.67+/-0.09, P<0.05). CONCLUSIONS: RPR101511A, which acts by inhibition of the PDGFr-TK, completely prevented angiographic loss of gain following PTCA and significantly reduced histological intimal hyperplasia. PMID- 10385506 TI - Additive effects of late preconditioning produced by monophosphoryl lipid A and the early preconditioning mediated by adenosine receptors and KATP channel. AB - BACKGROUND: The cardioprotective effect of preconditioning can be exerted within 1 to 2 hours after initial ischemia, termed classical or early preconditioning, or can reappear 24 hours later as second window or late preconditioning. The objective of this study was to study the interaction between late and early preconditioning and to determine the potential underlying mechanism. METHODS AND RESULTS: Adenosine receptor agonists and a KATP channel opener were used to achieve early preconditioning, and Monophosphoryl lipid A (MLA) was used to induce late preconditioning. Cultured chick ventricular myocytes were used as a myocyte model of simulated ischemia and preconditioning. Prior treatment of the myocyte with MLA caused a dose-dependent decrease in the ischemia-induced myocyte injury 24 hours later, consistent with a late preconditioning effect. L-NMMA, glibenclamide, or 5-hydroxydecanoic acid administered during the ischemia blocked the MLA effect. Twenty four hours after MLA treatment, a 5-minute exposure to ischemia, adenosine, adenosine A1 agonist CCPA, or A3 agonist resulted in less myocyte injury during the subsequent prolonged ischemia, as compared with cells pretreated with the vehicle and subsequently exposed to the same early preconditioning stimuli. In addition to its ability to enhance the early preconditioning effect by A1 and A3 agonists, MLA pretreatment also increased the phorbol ester- and pinacidil-mediated early preconditioning effect. CONCLUSIONS: This study defined a novel interaction in which the cardioprotective effect of early preconditioning is additive to that of late preconditioning and raised the possibility that both agents can be used as combined therapy in the treatment of ischemic heart disease. PMID- 10385507 TI - Thrombin-induced platelet activation is inhibited by high- and low-molecular weight heparin. AB - BACKGROUND: Thrombin binds to platelet glycoprotein Ib (Gp Ib), and this interaction contributes to platelet activation. Thrombin ligation to Gp Ib was recently shown to be inhibited by heparin, thus raising the hypothesis, investigated in this article, that heparin might inhibit thrombin-induced platelet activation. METHODS AND RESULTS: Aggregation of gel-filtered platelets by 1 nmol/L thrombin was reduced by both high-molecular-weight (MW) (14 500-Da) and low-MW (4500-Da) heparin, with IC50 values of 1.65+/-0.26 and 5.13+/-0.8 micromol/L, respectively. Homogeneous-MW fractions (16 000- to 13 000-Da range) were used to evaluate the heparin effect on intracytoplasmic calcium release by thrombin. Calcium mobilization by 1 nmol/L thrombin was reduced as a function of heparin concentration, and the inhibitory effect was correlated to the MW of heparin fractions (IC50 values were 1.9+/-0.39, 6.07+/-0.83, and 14. 8+/-0.43 micromol/L for 16 000-, 9000-, and 3000-Da heparin, respectively). Platelet aggregation and calcium mobilization by ADP and by the thrombin receptor activating peptide were not affected by heparin. The activation of Gp Ib-depleted platelets by alpha-thrombin was not inhibited by heparin. Moreover, platelet stimulation by heparin binding site phosphopyridoxylated thrombin, which has a severe impairment of Gp Ib ligation, was not affected by heparin. Finally, heparin did not interfere with the hydrolysis by thrombin of the protease activated receptor 1. CONCLUSIONS: These results demonstrated that heparin, by inhibiting the thrombin-Gp Ib interaction, is able to interfere with thrombin induced platelet activation. The extent of the inhibitory effect is directly related to the MW of heparin fractions. PMID- 10385508 TI - Urokinase receptor (uPAR, CD87) is a platelet receptor important for kinetics and TNF-induced endothelial adhesion in mice. AB - BACKGROUND: Urokinase plasminogen activator receptor (uPAR, CD87) is a widely distributed 55-kD, glycoprotein I-anchored surface receptor. On binding of its ligand uPA, it is known to increase leukocyte adhesion and traffic. Using genetically deficient mice, we explored the role of uPAR in platelet kinetics and TNF-induced platelet consumption. METHODS AND RESULTS: Anti-uPAR antibody stained platelets from normal (+/+) but not from uPAR-/- mice, as seen by fluorescence activated cell sorter analysis. 51Cr-labeled platelets from uPAR-/- donors survived longer than those from +/+ donors when injected into a +/+ recipient. Intratracheal TNF injection induced thrombocytopenia and a platelet pulmonary localization, pronounced in +/+ but absent in uPAR-/- mice. Aprotinin, a plasmin inhibitor, decreased TNF-induced thrombocytopenia. TNF injection markedly reduced the survival and increased the pulmonary localization of 51Cr-labeled platelets from +/+ but not from uPAR-/- donors, indicating that it is the platelet uPAR that is critical for their response to TNF. As seen by electron microscopy, TNF injection increased the number of platelets and polymorphonuclear neutrophils (PMNs) in the alveolar capillaries of +/+ mice, whereas in uPAR-/- mice, platelet trapping was insignificant and PMN trapping was slightly reduced. Platelets within alveolar capillaries of TNF-injected mice were activated, as judged from their shape, and this was evident in +/+ but not in uPAR-/- mice. CONCLUSIONS: These results demonstrate for the first time the critical role of platelet uPAR for kinetics as well as for activation and endothelium adhesion associated with inflammation. PMID- 10385509 TI - Images in Cardiovascular Medicine. Myocardial [18F]fluorodeoxyglucose uptake after heterotopic cardiac transplantation assessed by positron emission tomography. PMID- 10385510 TI - Platelet activation with unfractionated heparin at therapeutic concentrations and comparison with low-molecular-weight heparin and with a direct thrombin inhibitor. PMID- 10385511 TI - Paradoxical embolism. PMID- 10385512 TI - Rapport? PMID- 10385513 TI - Myocardial bridging. PMID- 10385514 TI - Physician noncompliance with the 1993 National Cholesterol Education Program (NCEP-ATPII) guidelines. PMID- 10385515 TI - Intracoronary ultrasound longitudinal reconstruction of a postangioplasty coronary artery dissection. PMID- 10385517 TI - Combined biochemical and electron microscopic analyses reveal the architecture of the mammalian U2 snRNP. AB - The 17S U2 small nuclear ribonucleoprotein particle (snRNP) represents the active form of U2 snRNP that binds to the pre-mRNA during spliceosome assembly. This particle forms by sequential interactions of splicing factors SF3b and SF3a with the 12S U2 snRNP. We have purified SF3b and the 15S U2 snRNP, an intermediate in the assembly pathway, from HeLa cell nuclear extracts and show that SF3b consists of four subunits of 49, 130, 145, and 155 kD. Biochemical analysis indicates that both SF3b and the 12S U2 snRNP are required for the incorporation of SF3a into the 17S U2 snRNP. Nuclease protection studies demonstrate interactions of SF3b with the 5' half of U2 small nuclear RNA, whereas SF3a associates with the 3' portion of the U2 snRNP and possibly also interacts with SF3b. Electron microscopy of the 15S U2 snRNP shows that it consists of two domains in which the characteristic features of isolated SF3b and the 12S U2 snRNP are conserved. Comparison to the two-domain structure of the 17S U2 snRNP corroborates the biochemical results in that binding of SF3a contributes to an increase in size of the 12S U2 domain and possibly induces a structural change in the SF3b domain. PMID- 10385516 TI - Large-scale chromatin unfolding and remodeling induced by VP16 acidic activation domain. AB - Analysis of the relationship between transcriptional activators and chromatin organization has focused largely on lower levels of chromatin structure. Here we describe striking remodeling of large-scale chromatin structure induced by a strong transcriptional activator. A VP16-lac repressor fusion protein targeted the VP16 acidic activation domain to chromosome regions containing lac operator repeats. Targeting was accompanied by increased transcription, localized histone hyperacetylation, and recruitment of at least three different histone acetyltransferases. Observed effects on large-scale chromatin structure included unfolding of a 90-Mbp heterochromatic chromosome arm into an extended 25-40 micrometers chromonema fiber, remodeling of this fiber into a novel subnuclear domain, and propagation of large-scale chromatin unfolding over hundreds of kilobase pairs. These changes in large-scale chromatin structure occurred even with inhibition of ongoing transcription by alpha-amanitin. Our results suggest a functional link between recruitment of the transcriptional machinery and changes in large-scale chromatin structure. Based on the observed long-range propagation of changes in large-scale chromatin structure, we suggest a possible rationale for the observed clustering of housekeeping genes within Mbp-sized chromosome bands. PMID- 10385518 TI - Assembly of 5S ribosomal RNA is required at a specific step of the pre-rRNA processing pathway. AB - A collection of yeast strains surviving with mutant 5S RNA has been constructed. The mutant strains presented alterations of the nucleolar structure, with less granular component, and a delocalization of the 25S rRNA throughout the nucleoplasm. The 5S RNA mutations affected helix I and resulted in decreased amounts of stable 5S RNA and of the ribosomal 60S subunits. The shortage of 60S subunits was due to a specific defect in the processing of the 27SB precursor RNA that gives rise to the mature 25S and 5.8S rRNA. The processing rate of the 27SB pre-rRNA was specifically delayed, whereas the 27SA and 20S pre-rRNA were processed at a normal rate. The defect was partially corrected by increasing the amount of mutant 5S RNA. We propose that the 5S RNA is recruited by the pre-60S particle and that its recruitment is necessary for the efficient processing of the 27SB RNA precursor. Such a mechanism could ensure that all newly formed mature 60S subunits contain stoichiometric amounts of the three rRNA components. PMID- 10385519 TI - Sli15 associates with the ipl1 protein kinase to promote proper chromosome segregation in Saccharomyces cerevisiae. AB - The conserved Ipl1 protein kinase is essential for proper chromosome segregation and thus cell viability in the budding yeast Saccharomyces cerevisiae. Its human homologue has been implicated in the tumorigenesis of diverse forms of cancer. We show here that sister chromatids that have separated from each other are not properly segregated to opposite poles of ipl1-2 cells. Failures in chromosome segregation are often associated with abnormal distribution of the spindle pole associated Nuf2-GFP protein, thus suggesting a link between potential spindle pole defects and chromosome missegregation in ipl1 mutant cells. A small fraction of ipl1-2 cells also appears to be defective in nuclear migration or bipolar spindle formation. Ipl1 associates, probably directly, with the novel and essential Sli15 protein in vivo, and both proteins are localized to the mitotic spindle. Conditional sli15 mutant cells have cytological phenotypes very similar to those of ipl1 cells, and the ipl1-2 mutation exhibits synthetic lethal genetic interaction with sli15 mutations. sli15 mutant phenotype, like ipl1 mutant phenotype, is partially suppressed by perturbations that reduce protein phosphatase 1 function. These genetic and biochemical studies indicate that Sli15 associates with Ipl1 to promote its function in chromosome segregation. PMID- 10385520 TI - Chromosomal influence on meiotic spindle assembly: abnormal meiosis I in female Mlh1 mutant mice. AB - In mouse oocytes, the first meiotic spindle is formed through the action of multiple microtubule organizing centers rather than a pair of centrosomes. Although the chromosomes are thought to play a major role in organizing the meiotic spindle, it remains unclear how a stable bipolar spindle is established. We have studied the formation of the first meiotic spindle in murine oocytes from mice homozygous for a targeted disruption of the DNA mismatch repair gene, Mlh1. In the absence of the MLH1 protein meiotic recombination is dramatically reduced and, as a result, the vast majority of chromosomes are present as unpaired univalents at the first meiotic division. The orientation of these univalent chromosomes at prometaphase suggests that they are unable to establish stable bipolar spindle attachments, presumably due to the inability to differentiate functional kinetochore domains on individual sister chromatids. In the presence of this aberrant chromosome behavior a stable first meiotic spindle is not formed, the spindle poles continue to elongate, and the vast majority of cells never initiate anaphase. These results suggest that, in female meiotic systems in which spindle formation is based on the action of multiple microtubule organizing centers, the chromosomes not only promote microtubule polymerization and organization but their attachment to opposite spindle poles acts to stabilize the forming spindle poles. PMID- 10385521 TI - Nuclear import of the TATA-binding protein: mediation by the karyopherin Kap114p and a possible mechanism for intranuclear targeting. AB - Binding of the TATA-binding protein (TBP) to the promoter is the first and rate limiting step in the formation of transcriptional complexes. We show here that nuclear import of TBP is mediated by a new karyopherin (Kap) (importin) family member, Kap114p. Kap114p is localized to the cytoplasm and nucleus. A complex of Kap114p and TBP was detected in the cytosol and could be reconstituted using recombinant proteins, suggesting that the interaction was direct. Deletion of the KAP114 gene led to specific mislocalization of TBP to the cytoplasm. We also describe two other potential minor import pathways for TBP. Consistent with other Kaps, the dissociation of TBP from Kap114p is dependent on RanGTP. However, we could show that double stranded, TATA-containing DNA stimulates this RanGTP mediated dissociation of TBP, and is necessary at lower RanGTP concentrations. This suggests a mechanism where, once in the nucleus, TBP is preferentially released from Kap114p at the promoter of genes to be transcribed. In this fashion Kap114p may play a role in the intranuclear targeting of TBP. PMID- 10385522 TI - A complex web of signal-dependent trafficking underlies the triorganellar distribution of P-selectin in neuroendocrine PC12 cells. AB - By analyzing the trafficking of HRP-P-selectin chimeras in which the lumenal domain of P-selectin was replaced with horseradish peroxidase, we determined the sequences needed for targeting to synaptic-like microvesicles (SLMV), dense core granules (DCG), and lysosomes in neuroendocrine PC12 cells. Within the cytoplasmic domain of P-selectin, Tyr777 is needed for the appearance of P selectin in immature and mature DCG, as well as for targeting to SLMV. The latter destination also requires additional sequences (Leu768 and 786DPSP789) which are responsible for movement through endosomes en route to the SLMV. Leu768 also mediates transfer from early transferrin (Trn)-positive endosomes to the lysosomes; i.e., operates as a lysosomal targeting signal. Furthermore, SLMV targeting of HRP-P-selectin chimeras, but not the endogenous SLMV protein synaptophysin/p38, previously shown to be delivered to SLMV directly from the plasma membrane, is a Brefeldin A-sensitive process. Together, these data are consistent with a model of SLMV biogenesis which involves an endosomal intermediate in PC12 cells. In addition, we have discovered that impairment of SLMV or DCG targeting results in a concomitant increase in lysosomal delivery, illustrating the entwined relationships between routes leading to regulated secretory organelles (RSO) and to lysosomes. PMID- 10385523 TI - Three v-SNAREs and two t-SNAREs, present in a pentameric cis-SNARE complex on isolated vacuoles, are essential for homotypic fusion. AB - Vacuole SNAREs, including the t-SNAREs Vam3p and Vam7p and the v-SNARE Nyv1p, are found in a multisubunit "cis" complex on isolated organelles. We now identify the v-SNAREs Vti1p and Ykt6p by mass spectrometry as additional components of the immunoisolated vacuolar SNARE complex. Immunodepletion of detergent extracts with anti-Vti1p removes all the Ykt6p that is in a complex with Vam3p, immunodepletion with anti-Ykt6p removes all the Vti1p that is complexed with Vam3p, and immunodepletion with anti-Nyv1p removes all the Ykt6p in complex with other SNAREs, demonstrating that they are all together in the same cis multi-SNARE complex. After priming, which disassembles the cis-SNARE complex, antibodies to any of the five SNARE proteins still inhibit the fusion assay until the docking stage is completed, suggesting that each SNARE plays a role in docking. Furthermore, vti1 temperature-sensitive alleles cause a synthetic fusion defective phenotype in our reaction. Our data show that vacuole-vacuole fusion requires a cis-SNARE complex of five SNAREs, the t-SNAREs Vam3p and Vam7p and the v-SNAREs Nyv1p, Vti1p, and Ykt6p. PMID- 10385524 TI - Molecular characterization of caveolin association with the Golgi complex: identification of a cis-Golgi targeting domain in the caveolin molecule. AB - Caveolins are integral membrane proteins which are a major component of caveolae. In addition, caveolins have been proposed to cycle between intracellular compartments and the cell surface but the exact trafficking route and targeting information in the caveolin molecule have not been defined. We show that antibodies against the caveolin scaffolding domain or against the COOH terminus of caveolin-1 show a striking specificity for the Golgi pool of caveolin and do not recognize surface caveolin by immunofluorescence. To analyze the Golgi targeting of caveolin in more detail, caveolin mutants were expressed in fibroblasts. Specific mutants lacking the NH2 terminus were targeted to the cis Golgi but were not detectable in surface caveolae. Moreover, a 32-amino acid segment of the putative COOH-terminal cytoplasmic domain of caveolin-3 was targeted specifically and exclusively to the Golgi complex and could target a soluble heterologous protein, green fluorescent protein, to this compartment. Palmitoylation-deficient COOH-terminal mutants showed negligible association with the Golgi complex. This study defines unique Golgi targeting information in the caveolin molecule and identifies the cis Golgi complex as an intermediate compartment on the caveolin cycling pathway. PMID- 10385525 TI - Integrin-mediated activation of focal adhesion kinase is required for signaling to Jun NH2-terminal kinase and progression through the G1 phase of the cell cycle. AB - The extracellular matrix exerts a stringent control on the proliferation of normal cells, suggesting the existence of a mitogenic signaling pathway activated by integrins, but not significantly by growth factor receptors. Herein, we provide evidence that integrins cause a significant and protracted activation of Jun NH2-terminal kinase (JNK), while several growth factors cause more modest or no activation of this enzyme. Integrin-mediated stimulation of JNK required the association of focal adhesion kinase (FAK) with a Src kinase and p130(CAS), the phosphorylation of p130(CAS), and subsequently, the recruitment of Crk. Ras and PI-3K were not required. FAK-JNK signaling was necessary for proper progression through the G1 phase of the cell cycle. These findings establish a role for FAK in both the activation of JNK and the control of the cell cycle, and identify a physiological stimulus for JNK signaling that is consistent with the role of Jun in both proliferation and transformation. PMID- 10385527 TI - Thin filament protein dynamics in fully differentiated adult cardiac myocytes: toward a model of sarcomere maintenance. AB - Sarcomere maintenance, the continual process of replacement of contractile proteins of the myofilament lattice with newly synthesized proteins, in fully differentiated contractile cells is not well understood. Adenoviral-mediated gene transfer of epitope-tagged tropomyosin (Tm) and troponin I (TnI) into adult cardiac myocytes in vitro along with confocal microscopy was used to examine the incorporation of these newly synthesized proteins into myofilaments of a fully differentiated contractile cell. The expression of epitope-tagged TnI resulted in greater replacement of the endogenous TnI than the replacement of the endogenous Tm with the expressed epitope-tagged Tm suggesting that the rates of myofilament replacement are limited by the turnover of the myofilament bound protein. Interestingly, while TnI was first detected in cardiac sarcomeres along the entire length of the thin filament, the epitope-tagged Tm preferentially replaced Tm at the pointed end of the thin filament. These results support a model for sarcomeric maintenance in fully differentiated cardiac myocytes where (a) as myofilament proteins turnover within the cell they are rapidly exchanged with newly synthesized proteins, and (b) the nature of replacement of myofilament proteins (ordered or stochastic) is protein specific, primarily affected by the structural properties of the myofilament proteins, and may have important functional consequences. PMID- 10385526 TI - The zinc finger protein A20 inhibits TNF-induced NF-kappaB-dependent gene expression by interfering with an RIP- or TRAF2-mediated transactivation signal and directly binds to a novel NF-kappaB-inhibiting protein ABIN. AB - The zinc finger protein A20 is a tumor necrosis factor (TNF)- and interleukin 1 (IL-1)-inducible protein that negatively regulates nuclear factor-kappa B (NF kappaB)-dependent gene expression. However, the molecular mechanism by which A20 exerts this effect is still unclear. We show that A20 does not inhibit TNF- induced nuclear translocation and DNA binding of NF-kappaB, although it completely prevents the TNF- induced activation of an NF-kappaB-dependent reporter gene, as well as TNF-induced IL-6 and granulocyte macrophage-colony stimulating factor gene expression. Moreover, NF-kappaB activation induced by overexpression of the TNF receptor-associated proteins TNF receptor-associated death domain protein (TRADD), receptor interacting protein (RIP), and TNF recep- tor-associated factor 2 (TRAF2) was also inhibited by expression of A20, whereas NF-kappaB activation induced by overexpression of NF-kappaB-inducing kinase (NIK) or the human T cell leukemia virus type 1 (HTLV-1) Tax was unaffected. These results demonstrate that A20 inhibits NF-kappaB-dependent gene expression by interfering with a novel TNF-induced and RIP- or TRAF2-mediated pathway that is different from the NIK-IkappaB kinase pathway and that is specifically involved in the transactivation of NF-kappaB. Via yeast two-hybrid screening, we found that A20 binds to a novel protein, ABIN, which mimics the NF-kappaB inhibiting effects of A20 upon overexpression, suggesting that the effect of A20 is mediated by its interaction with this NF-kappaB inhibiting protein, ABIN. PMID- 10385530 TI - Overview of sleep disorders: where does obstructive sleep apnea syndrome fit in? AB - Otolaryngologists deal primarily with the disorders of obstructive sleep apnea and primary snoring. It is important to realize that although these two disorders are common in the general population, they make up only a small segment of the entire field of sleep disorders medicine. This article attempts to introduce the otolaryngologist to the complexity of this field, help to gain respect and understanding of those practitioners dealing with this entire field, and learn why there is such a broad appeal of this field of medicine to so many subspecialists. Also presented are a table describing the classification of sleep disorders and a short tribute to those individuals who founded this relatively new field of medicine. PMID- 10385531 TI - Sleep studies. Current techniques and future trends. AB - Diagnosis of obstructive sleep apnea has been termed a laboratory diagnosis rather than a clinical diagnosis because one may not be able to make the diagnosis based on the history and physical examination alone. The polysomnogram was developed to give clinicians and researchers objective data on physiologic events occurring during the patient's sleep. From this, obstructive breathing patterns can be diagnosed and if pathologic, appropriate treatment can be instituted. Although the polysomnogram has been the gold standard for diagnosis for more than two decades, it is an expensive and time-consuming procedure. Current technologies for polysomnogram are reviewed, as well as proposals for alternatives that may be more cost and time effective. PMID- 10385529 TI - The dynamics of protein kinase B regulation during B cell antigen receptor engagement. AB - This study has used biochemistry and real time confocal imaging of green fluorescent protein (GFP)-tagged molecules in live cells to explore the dynamics of protein kinase B (PKB) regulation during B lymphocyte activation. The data show that triggering of the B cell antigen receptor (BCR) induces a transient membrane localization of PKB but a sustained activation of the enzyme; active PKB is found in the cytosol and nuclei of activated B cells. Hence, PKB has three potential sites of action in B lymphocytes; transiently after BCR triggering PKB can phosphorylate plasma membrane localized targets, whereas during the sustained B cell response to antigen, PKB acts in the nucleus and the cytosol. Membrane translocation of PKB and subsequent PKB activation are dependent on BCR activation of phosphatidylinositol 3-kinase (PI3K). Moreover, PI3K signals are both necessary and sufficient for sustained activation of PKB in B lymphocytes. However, under conditions of continuous PI3K activation or BCR triggering there is only transient recruitment of PKB to the plasma membrane, indicating that there must be a molecular mechanism to dissociate PKB from sites of PI3K activity in B cells. The inhibitory Fc receptor, the FcgammaRIIB, mediates vital homeostatic control of B cell function by recruiting an inositol 5 phosphatase SHIP into the BCR complex. Herein we show that coligation of the BCR with the inhibitory FcgammaRIIB prevents membrane targeting of PKB. The FcgammaRIIB can thus antagonize BCR signals for PKB localization and prevent BCR stimulation of PKB activity which demonstrates the mechanism for the inhibitory action of the FcgammaRIIB on the BCR/PKB response. PMID- 10385528 TI - Direct involvement of ezrin/radixin/moesin (ERM)-binding membrane proteins in the organization of microvilli in collaboration with activated ERM proteins. AB - Ezrin/radixin/moesin (ERM) proteins have been thought to play a central role in the organization of cortical actin-based cytoskeletons including microvillar formation through cross-linking actin filaments and integral membrane proteins such as CD43, CD44, and ICAM-2. To examine the functions of these ERM-binding membrane proteins (ERMBMPs) in cortical morphogenesis, we overexpressed ERMBMPs (the extracellular domain of E-cadherin fused with the transmembrane/cytoplasmic domain of CD43, CD44, or ICAM-2) in various cultured cells. In cultured fibroblasts such as L and CV-1 cells, their overexpression significantly induced microvillar elongation, recruiting ERM proteins and actin filaments. When the ERM binding domains were truncated from these molecules, their ability to induce microvillar elongation became undetectable. In contrast, in cultured epithelial cells such as MTD-1A and A431 cells, the overexpression of ERMBMPs did not elongate microvilli. However, in the presence of EGF, overexpression of ERMBMPs induced remarkable microvillar elongation in A431 cells. These results indicated that ERMBMPs function as organizing centers for cortical morphogenesis by organizing microvilli in collaboration with activated ERM proteins. Furthermore, immunodetection with a phosphorylated ERM-specific antibody and site-directed mutagenesis suggested that ERM proteins phosphorylated at their COOH-terminal threonine residue represent activated ERM proteins. PMID- 10385532 TI - Disordered breathing during sleep in newborns, infants, and children. Symptoms, diagnosis, and treatment. AB - Although the polysomnographic findings of sleep-disordered breathing in children are similar to those in adults, the underlying causes will vary significantly from adults, depending on whether one is dealing with a newborn, infant, or child. How they react to the disease process is also at times different than seen in the adult and subsequent testing and treatment will also vary considerably. These differences and similarities are reviewed in this article. PMID- 10385533 TI - Complications of snoring, upper airway resistance syndrome, and obstructive sleep apnea syndrome in adults. AB - The complications of sleep-disordered breathing can be separated into two categories. First are those disorders that primarily are brought on by the sleep disorder itself. The second category is those pre-existing medical problems that are aggravated by the sleep disorder. This article examines the consequences of obstructive breathing disorders during sleep and reviews some of the current theories as to the pathophysiology of those problems directly resulting from the sleep disorder. PMID- 10385534 TI - Oral and maxillofacial surgery for the management of obstructive sleep apnea syndrome. AB - The initial reports of treating obstructive sleep apnea using the uvulopalatopharyngoplasty were encouraging; however, as further trials of this procedure were reported, it began to show disappointing results. It was found that the retropalatal airway was not the only site of obstruction and procedures would need to be developed that would address obstruction in the other portions of the airway involved, notably in the retrolingual or hypopharyngeal portion of the airway. It was first reported by oral surgeons that mandibular surgery could also improve sleep apnea and through their work and the work of others, techniques have been developed using skeletal surgery to enhance the patency of the airway during sleep. This article describes some of these techniques and their indications, complications, and results. PMID- 10385535 TI - The nasal airway and obstructed breathing during sleep. AB - Nasal obstruction whether partial or complete can influence the quality of sleep and has been strongly linked to the genesis of obstructed breathing during sleep (OBS). The relationship between nasal airflow and the process of upper airway collapse is complex. The first part of this article reviews the nasal anatomy with an emphasis on the sites of nasal obstruction, the effect of the nasal reflexes on the pulmonary system, and the pathophysiology of the development of OBS. The second part reviews the common causes of increased nasal resistance, the assessment of nasal passages, and the treatment options. This article also includes literature in support of and refuting the postulated mechanisms by which nasal obstruction can effect the respiratory system during sleep. PMID- 10385536 TI - Oropharyngeal surgery in the management of upper airway obstruction during sleep. AB - In the surgical management of snoring and sleep apnea, surgery to the oropharynx was the initial procedure used to treat sleep-related disorders. This article reviews both the various procedures available for this and the benefits and drawbacks of these procedures so the practitioner may be able to choose which type would be most beneficial for a particular patient. PMID- 10385537 TI - Suspension sutures for the treatment of obstructive sleep apnea and snoring. AB - Treatment of airway collapse in the retrolingual airway for obstructive sleep apnea syndrome and snoring has been a frequently frustrating exercise. There are several procedures that have been used with varying degrees of success for some time. These procedures include genioglossus advancement and hyoid suspension, as well as various forms of lingual plasty and lingual reduction. A new technique was introduced at the 1998 meeting of the American Academy of Otolaryngology-Head and Neck Surgery in San Antonio that consisted of using a suspension screw to support the hypopharyngeal soft tissues, specifically, the base of the tongue, to prevent its posterior displacement during sleep. Some of the initial results of these studies have been promising and are reviewed here. PMID- 10385538 TI - Effects of drugs on sleep AB - Many commonly prescribed medications and substances of abuse can have significant effects on sleep and wakefulness. Chronic use or abuse of certain drugs may lead to the development of substance-related sleep disorders. Primary sleep disorders, such as apnea, periodic movement disorders, and parasomnias, may be exacerbated by various drugs. This article summarizes the effects of widely used medications and recreational drugs on sleep. PMID- 10385539 TI - Recognition and consequences of obstructive sleep apnea hypopnea syndrome AB - There is a growing recognition of sleep-disordered breathing (SDB) in patient groups and in the general population. This article reviews issues related to recognizing the disorder, including the problems of relying on narrowly defined polysomnographic data for case findings and for assessment of disease severity. The distributions of symptoms and physiologic measurements of SDB in the population and their inter-relationships are reviewed. The epidemiological data that address risk factors and consequences of sleep apnea hypopnea syndrome (SAHS) also are discussed, with recommendations regarding recognition priorities. PMID- 10385540 TI - Decision making in obstructive sleep-disordered breathing: putting It all together AB - Obstructive sleep-disordered breathing consists of a spectrum ranging from the upper airway resistance syndrome to complete apnea. Although this disorder is relatively common, it is still under-recognized, resulting in a significant increase in morbidity and mortality. This article describes the rationale for treating this disorder. Then, using the best available evidence, develops a systemic approach to the problem, covering recognition, diagnosis, and treatment. PMID- 10385541 TI - Current and future methodology for monitoring sleep AB - Sleep is much more than simply decreased consciousness, and the distinctions among the stages of non-REM and REM sleep are of significance for the polysomnographer because of their implications for physiologic events. The rules for staging sleep are essentially unchanged since their original description in 1968. This article describes this system for sleep staging, some of its weaknesses, and potential new approaches. PMID- 10385542 TI - A look toward the future AB - There will be many changes in the sleep field in the next 5 to 10 years. These will include increments in our knowledge of basic neurobiologic mechanisms driving sleep and the impact of sleep loss on general health. The technology used in the sleep laboratory will likely change as well, leading to a larger range of available tests and new ways to conduct standard ones. Finally, as the knowledge base in sleep increases, the expertise required to practice sleep medicine will rise, leading to a better trained, more focused practitioner. PMID- 10385543 TI - The importance of grading in endometrial cancer. PMID- 10385544 TI - An analysis of two versus three grades for endometrial carcinoma. AB - Introduction. The current grading of uterine endometrioid adenocarcinoma utilizes a three-grade system based on the amount of nonsquamous solid histologic architecture. Of these three grades, we questioned the practical clinical utility of the intermediate grade. Methods. We retrospectively reviewed endometrial biopsy and uterine histology specimens, quantifying the percentage amount of nonsquamous solid tumor by intervals of 10. We then compared these percentage values to other histopathologic prognostic variables. Results. Eighty-five Stage I and II endometrioid adenocarcinoma patients had their preoperative endometrial curettings and operative hysterectomy pathology reviewed independently by two gynecologic pathologists for surgical staging and outcome with a mean follow-up of 6 years. Using a two-tiered system for assessing uterine tumor grade with a delineating value of 20% nonsquamous solid tumor, we found less interobserver variation (kappa = 0.966) compared to the current three-tiered grading system (kappa = 0.526). There were no differences between the two- and three-tiered grading systems regarding myometrial invasion, lymph vascular space invasion, and survival. In the diagnosis of endometrial biopsies, the two-tiered system also improved the prediction of uterine histology grade over the three-tiered system, 90 and 63%, respectively. Conclusion. A two-grade architecture system with a delineation value of 20% would be more reliable and less cumbersome and would have the same or better prognostic significance as the currently used three-grade system. PMID- 10385545 TI - Pelvic and paraortic lymph nodal status in advanced ovarian cancer and survival. AB - BACKGROUND: In order to analyze the prognostic role of node involvement in advanced ovarian cancer, we have analyzed data from a randomized clinical trial on advanced ovarian cancer. METHODS: Cases were 456 women who entered a randomized multicentric clinical trial comparing two cisplatin-based schemes of treatment after cytoreductive surgery for advanced stage III-IV ovarian cancer. They underwent selective pelvic and/or paraortic lymphadenectomy. RESULTS: A total of 161 (35.3%) cases had positive nodes. The frequency of positive nodes was statistically significantly higher in FIGO stage IV than in stage III. Also grade 3 tumors were more likely to have positive nodes than grade 1-2 tumors. No association was observed between nodal status and response to chemotherapy. The 3 year survival was 46.2 (standard error (SE) = 3.4 based on 147 deaths) and 44.6 (SE = 4.4, based on 84 deaths), respectively, in negative and positive node groups. The corresponding values, when the analysis was performed considering only subjects with residual tumor <1 cm or absent, after first-line cytoreductive surgery were 66.2 (SE = 5.7) and 62.4 (SE = 9.6). CONCLUSIONS: We did not find any association between nodal status and survival. Particularly, nodal status was not a prognostic factor for survival in the subgroup of women with residual tumor <1 cm or absent after cytoreductive surgery. PMID- 10385546 TI - Assessment of biological variation and analytical imprecision of CA 125, CEA, and TPA in relation to monitoring of ovarian cancer. AB - OBJECTIVES: Changes in serial tumor marker results during monitoring of patients with ovarian cancer are due not only to deterioration or amelioration of the patient's condition, but also to preanalytical sources of variation (CPP), total random analytical error, and within-subject normal biological variation. The aim of the study was to assess (i) the analytical imprecision (CVA) and the average inherent intra- and interindividual biological variation (CVTI and CVG, respectively) for CA 125, CEA, and TPA in a group of healthy women; (ii) the significance of changes in serial results of each marker; and (iii) the index of individuality. METHODS: The study group consisted of 31 healthy women. Sixteen blood samples from each subject were collected in four series over a period of approximately 1 year. Data analysis was based on ANOVA. The index of individuality was calculated as ((CV2A + CV2TI)/CV2G)1/2 and the critical difference for a change between two consecutive concentrations as radical2xZx(CV2P + CV2A + CV2TI)1/2 (Z = 1.65 for unidirectional and 1.96 for bidirectional changes, P A transition at nt 7520 (100%, 27/27), A-->C transition at nt 7729 (70%; 19/27), and G-->A transition at nt 7841 (78%; 21/27). Selective mutations were observed at the YY1 binding sites of HPV-16 URR in the 3 patients with invasive cervical cancer who have the episomal forms of HPV-16 DNA: A-->C transition at nt 7484 and G-->A transition at nt 7488 (YY1-binding site 2; from 7481 to 7489). Additionally, C-->T transition at nt 7785 (YY1-binding site 3; from 7781 to 7790) was found in 2 of 3 patients. No YY1 site mutations were detected in the 12 CIN patients and in the HPV-integrated invasive cancer patients. To determine whether these mutations have effects on the expression of HPV E6/E7 genes driven by URR, the transient transfection assay was employed using URR-CAT reporter plasmid. The relative activities of three URR mutants from episomal HPV-16 DNA of cervical cancers were two- to fourfold higher than that of the HPV-16 URR prototype. In contrast, the URRs from integrated HPV-16 DNA in cervical cancer and from episomal HPV-16 DNA in CIN, where no mutation of the YY1 binding site was detected, showed similar levels of promoter activity to that of the URR prototype. CONCLUSIONS: Our results support the hypothesis that the mutation at the YY1 binding site is functionally related to the development of cervical neoplasia caused by episomal HPV-16 DNA in Korean cervical cancer patients. Thus, mutation in the YY1 site of episomal HPV-16 URR may play a corresponding role of HPV integration in the progression of cervical cancer. PMID- 10385548 TI - Pilot study of concurrent cisplatin, 5-fluorouracil, and external beam radiotherapy prior to radical surgery +/- intraoperative electron beam radiotherapy in locally advanced cervical cancer. AB - PURPOSE: The purpose of this study was to describe the feasibility of a combined preoperative chemoradiation program followed by radical surgery in advanced cervical cancer. MATERIALS AND METHODS: From February 1988 to April 1997, 40 patients with carcinoma of the cervix were treated with preoperative external beam radiotherapy to 45 Gy in 5 weeks. Patients received concurrent continuous infusion cisplatin (20 mg/m2) and 5-fluorouracil (1500 mg) chemotherapy during the first (days 1-4) and fifth (days 22-25) weeks of the radiation course. Radical surgery was performed 4-6 weeks after the completion of the preoperative treatment. Intraoperative radiotherapy was given to 20 patients, based on intraoperative assessment. RESULTS: Toxicity associated with chemoradiation was usually mild except in two patients who presented WHO grade 4 bone marrow aplasia. Three patients developed postoperative ureterovaginal fistula, and five patients developed long-term hydronephrosis that needed ureteral stenting. Clinical response was observed in 95% of the patients (55% complete response). The analysis of the surgical specimens revealed complete pathological response in 67.5% of the cases and partial pathological response in 32.5%. As expected, the degree of pathological response was predicted by the degree of clinical response (P = 0.001). Nine-year local control, distant metastases-free survival, disease free survival, and overall survival were 86, 84, 81, and 85%, respectively. Patients displaying a complete pathological response had statistically significant improved local control (P = 0.004), distant metastases-free survival (P = 0.009), disease-free survival (P = 0.002), and overall survival (P = 0.038). CONCLUSIONS: Cisplatin plus 5-fluorouracil preoperative chemoradiation is active and usually well tolerated in locally advanced carcinoma of cervix, inducing a high rate of clinical and pathological complete responses. When this therapy is followed by radical surgery, the local control rates are excellent, even in patients with advanced stages or poor response. These improved local control rates may be achievable only through extensive surgical resection, with a parallel increase in the complication rates. PMID- 10385549 TI - Expression of estrogen receptor alpha mRNA and protein variants in human endometrial carcinoma. AB - Breast cancer tissue has been shown to contain alternatively spliced estrogen receptor alpha (ER-alpha) mRNA variants, which have altered biological activities compared to the full-length ER-alpha. The development of endometrial cancer, as well as drug resistance in breast cancer patients undergoing tamoxifen therapy, may represent altered ER-alpha function secondary to specific exon deletions. While the literature is replete with ER mRNA variant data, little information is available regarding the presence and function of endometrial ER variant proteins. We evaluated the presence of human ER-alpha mRNA and protein variants in six premenopausal, six postmenopausal, and six endometrial carcinoma samples. Reverse transcription-polymerase chain reaction, DNA hybridization, and sequencing techniques identified exon 4, exon 5, and exon 7 mRNA splice variants in all patients as well as MCF-7 and Ishikawa cell lines. Presence of translated proteins for full-length ER-alpha, as well as splice variants, was investigated by Western blot analysis using antibodies directed against the N-terminus, hinge region, and C-terminus portions of the ER. These experiments confirmed the presence of immunopositive protein bands of approximately 64-66 kDa in all patients corresponding to wild-type ER-alpha. A protein band migrating at 41 kDa, consistent with an exon 5 splice variant, was only seen in endometrial adenocarcinoma samples. Premenopausal and postmenopausal endometrial samples did not contain detectable amounts of ER splice variant protein. Human ER-alpha mRNA variants are present in all human endometrial samples, but detectable levels of variant proteins are only observed in patients with endometrial adenocarcinoma. PMID- 10385550 TI - Intraperitoneal alpha-interferon alternating with cisplatin in residual ovarian carcinoma: a phase II Gynecologic Oncology Group study. AB - OBJECTIVE: The aim of this study was to study the combination of intraperitoneal alpha-interferon and cisplatin administered second-line in an alternating sequence in small volume residual epithelial ovarian cancer after second-look surgery and the activity of this combination based on prior response to first line platinum compounds. METHODS: Sixty-two patients with minimal residual (<0.5 cm) epithelial ovarian cancer at reassessment laparotomy were entered into a multicenter trial of intraperitoneal alpha-interferon alternating with cisplatin given for eight cycles unless disease progression or unacceptable toxicity occurred. The patients were considered favorable if they were platinum-sensitive and/or relapsed 6 months or longer after completing treatment. Another reassessment laparotomy was performed within 12 weeks of completion of treatment in patients who were in clinical remission. RESULTS: Fifty-four patients were clinically evaluable and 18 were surgically reassessed, 5 of whom had a negative reassessment operation (20% complete response and 8% partial response). Of the 54 patients evaluable for toxicity, the most common adverse effects of more than grade 2 were gastrointestinal in 13 (47%), neutropenia in 9 (17%), and leukopenia in 6 (12%). Grade 4 toxicity was seen in 10 instances: 4 gastrointestinal, 2 neutropenia, 2 thrombocytopenia, 1 wound infection, and 1 allergic reaction. CONCLUSIONS: alpha-Interferon and cisplatin are active agents in favorable patients with minimal residual epithelial ovarian cancer at second-look. The combination of the two drugs administered in an alternating sequence appears to be associated with more side effects than when either drug is administered alone. The combination produced response rates similar to those seen when either drug is given alone. PMID- 10385552 TI - Squamous cell carcinoma of the vulva in Brazil: prognostic importance of host and viral variables. AB - BACKGROUND: Certain clinicopathologic features of vulvar squamous cell carcinoma have been correlated with adverse prognosis. However, few large-scale studies have addressed their role in patient survival. This study examined the relationship between multiple variables and prognosis in a large group of vulvar cancers in Brazil. METHODS: One hundred eighty-four Brazilian women with vulvar carcinoma were studied and the following variables recorded: age, pathologic TNM stage, survival, histologic grade, tumor histologic pattern, invasion pattern, tumor thickness, and tissue stromal and inflammatory response. Human papillomavirus (HPV) was detected by polymerase chain reaction amplification of extracted archival DNA. Data were analyzed using Cox proportional hazards modeling. RESULTS: After controlling for age, the probability of cancer survival decreased with increasing age, stage, grade, and tumor thickness, a fibromyxoid stromal response, infiltrative growth pattern, and basaloid histologic pattern. With the exception of fibromyxoid stromal response, each of these variables remained prognostically significant after adjustment for several other predictors in a multivariate model. Women whose tumors displayed a basaloid pattern were 3.5 times as likely to die from cancer than those with keratinizing tumors [hazard ratio (HR) = 3.5, 95% CI(1.3-9.2)]. An infiltrative invasion pattern strongly increased the probability of cancer death [HR = 4.6, 95% CI(1.9,11.4)]. HPV status did not influence survival, despite its association with basaloid histology. CONCLUSIONS: Previously reported associations of negative HPV status and fibromyxoid response with adverse prognosis in vulvar cancer were not confirmed by multivariate analysis. Basaloid variants, and particularly diffusely infiltrative tumors, carry an adverse prognosis. PMID- 10385551 TI - An immunohistochemical analysis of heat shock protein 70, p53, and estrogen receptor status in carcinoma of the uterine cervix. AB - OBJECTIVES: It has been shown that heat shock proteins (HSPs) protect cells from death caused by various noxious stimuli. Overexpression of HSP70 seems to be related to hormonal regulation of cell proliferation and/or down-regulation of sex steroid receptors. Wild-type p53 has been reported to repress HSP70 gene expression. It has been shown that mutant p53-HSP70 complex is highly expressed in cancer. However, the relationship between HSPs and steroid receptors or tumor suppressor gene products has not been well understood in uterine cervical carcinoma. This study was undertaken to examine the expression of HSP70, estrogen receptor (ER), and p53 in carcinoma of the uterine cervix. In addition, we analyzed HPV infection status and compared it to such immunohistochemical parameters. We also analyzed the relationship between these biological products and their clinicopathologic characteristics. METHODS: Paraffin-embedded tissue sections were obtained from 84 patients with carcinoma of the uterine cervix. Expression of HSP70, p53, and ER was evaluated by immunohistochemical staining using anti-HSP70 monoclonal antibody (SPA810), anti-p53 (BP53.12), and ER1D5 antibody, respectively. PCR HPV detection was done by dot hybridization method. RESULTS: Positive staining of HSP70 was detected in 73% of the cases. HSP70 positivity was significantly higher in stage I cervical cancer than in stages II IV (P = 0.02). This was associated with neither tumor size, lymph node status, parametrial involvement status, nor tumor markers (TA-4). Furthermore, there was no significant correlation between HSP70 positivity and the expression of p53 or ER or HPV infection status. CONCLUSION: These data suggested that HSP70 positivity was frequent in uterine cervical cancer, especially in the early stages. However, this was not significantly correlated with clinicopathologic characteristics nor with the expression of p53 or ER nor with HPV infection in carcinoma of the uterine cervix. PMID- 10385553 TI - Corticosteroids in the management of bowel obstruction on a gynecological oncology unit. AB - The development of gastrointestinal obstruction commonly occurs as a complication of advanced gynecological cancer. While surgery remains the mainstay of treatment for these patients, it is not always feasible, and when it is performed, it does not always resolve the obstruction. In this prospective study of patients presenting to a gynecologic oncology unit, 13 patients were administered 8 mg of dexamethasone subcutaneously or intravenously for a minimum of 3 days to manage the symptoms of bowel obstruction. Nine patients (69%) had a response to this therapy with decreased pain, nausea, and vomiting and improved oral intake. This response was maintained for a median of 31 days, with 7 of the 9 patients maintaining this symptomatic response until death. Mean survival of those responding was 39 days, including a subgroup of patients with extremely limited prognosis who, at their request, were discharged from the hospital in order to die at home. This subgroup had a mean survival of 20 days. The mean survival for nonresponders was 54 days. In patients for whom surgery is not contemplated, corticosteroids may provide a palliative treatment for bowel obstruction secondary to malignancy, provided there are no contraindications. PMID- 10385554 TI - The significance of adnexal involvement in endometrial carcinoma. AB - OBJECTIVE: To evaluate the prognostic significance of and predictive factors for adnexal involvement (AI) in patients with endometrial carcinoma. METHODS: We retrospectively reviewed the pathological features and outcomes of endometrial carcinoma patients. The prognostic significance of AI was examined by univariate and multivariate analyses. Median follow-up was 30.7 months. RESULTS: Of the 382 cases reviewed, 40 (10.5%) had AI. Patients with AI had a worse 5-year disease free (DFS) survival (73.1 vs 37.1%, P < 0.0001) than patients without AI. However, patients with AI had multiple adverse features, including high grade disease, lymphovascular invasion, and additional sites of extrauterine disease. After controlling for these factors on multivariate analysis, AI lost its prognostic significance (P = 0.56). The 12 AI patients without other extrauterine disease had a favorable outcome (5-year DFS of 70.9%). Factors predictive of AI on logistic regression were metastatic disease, positive peritoneal washings, cervical involvement, and unfavorable histology. CONCLUSION: Endometrial carcinoma patients with AI have relatively poor prognoses. However, AI per se has little, if any, independent prognostic significance. The poor outcomes seen in these patients appear to result from the preponderance of other adverse pathologic factors. PMID- 10385555 TI - Survival of women with surgical stage II endometrial cancer. AB - OBJECTIVE: The aim of this study was to report survival and determine prognostic factors and results of therapy in women with surgical stage II endometrial cancer. METHODS: Forty-eight consecutive women with surgical stage II endometrial cancer treated at the University of Vermont between March 1984 and March 1998 were reviewed. Patients' characteristics, surgical procedure, postoperative treatment and its complications, and tumor recurrence and its treatment were recorded. In addition, a formal review of their pathological material for confirmation of the diagnosis was performed. RESULTS: The median duration of follow-up was 6.2 years. Three patients (6.3%) had tumor recurrence and two (4.2%) died of their disease. The estimated 5-year overall survival and disease free survivals were 92.1% (SE = 5.5%, 95% confidence interval: 81.3, 100%) and 89.9% (SE = 5.8%, 95% confidence interval: 78.5%, 100%), respectively. None of the patients treated by total abdominal hysterectomy followed by both whole pelvic and vaginal cuff radiation therapy (the main line of treatment for patients in whom cervical involvement was diagnosed following hysterectomy, n = 20) or by radical hysterectomy (the main line of treatment for patients in whom cervical involvement was known before hysterectomy, n = 11) had tumor recurrence. Three of 17 (17.6%) patients treated with total abdominal hysterectomy followed by either whole pelvic (n = 13) or vaginal cuff (n = 4) radiation therapy had tumor recurrence. The difference between those two groups was statistically significant (0/31 versus 3/17, P = 0.02). There was no difference in survival among women with stage IIA and IIB or women who underwent radical abdominal hysterectomy and those who underwent total abdominal hysterectomy with postoperative pelvic and vaginal cuff radiation. Morbidity secondary to therapy was mild. Age, depth of myometrial invasion, tumor histology, and grade were not significantly related to recurrence. CONCLUSIONS: Survival of women with surgical stage II endometrial cancer is excellent especially among those treated with total abdominal hysterectomy followed by both pelvic and vaginal cuff radiotherapy or by radical abdominal hysterectomy. PMID- 10385556 TI - Clonal expansion of T cells that are specific for autologous ovarian tumor among tumor-infiltrating T cells in humans. AB - OBJECTIVE: The purpose of this study was to determine whether oligoclonally expanding tumor-infiltrating lymphocytes (TIL) were tumor-specific. STUDY DESIGN: Peripheral blood lymphocytes (PBL) from an ovarian tumor-bearing patient were stimulated in vitro with an autologous cancer cell line (SMOV-2). Then genes coding for the third complementarity-determining region of the T cell receptor (TCR) beta chain were amplified by reverse transcription-polymerase chain reaction and separated by single-strand conformation polymorphism. Accumulated TCR clonotypes in vitro and in vivo in TIL were compared. RESULTS: Clonal expansion of T cells was generated from PBL by stimulation with SMOV-2. A portion of the proliferated clonotypes was found to be identical to those accumulated in TIL in vivo. CONCLUSION: This is the first demonstration that accumulating T cell clones in TIL recognize antigen(s) on an autologous tumor. Further characterization of such T cell clonotypes may lead to tumor antigen-specific immunotherapy. PMID- 10385557 TI - Secretion of activin A in recurrent epithelial ovarian carcinoma. AB - OBJECTIVES: Activin A is a dimeric protein, composed of two beta-A subunits, that belongs to the TGF-beta family of growth factors. Most primary epithelial ovarian tumors (96%) synthesize and secrete activin protein in vitro and preliminary studies show that serum levels of activin are frequently elevated in women with epithelial ovarian cancer. Our objectives were to expand on studies of serum activin A levels in women with epithelial ovarian cancer and to determine whether levels of activin A correlate with the clinical course of disease. METHOD: Preoperative serum activin A levels were measured in 41 patients with epithelial ovarian cancer. In addition, serum activin A levels were measured in all available postoperative samples from the subset of these patients (n = 26) who had an elevated preoperative serum activin A level. Medical record information was used to compare each patient's serum levels of activin A to the clinical course of disease. RESULTS: Seventy-two percent of the stage III and IV patients (26/36), and none (0/5) of the stage I patients, had an elevated preoperative serum activin level. In postoperative samples, activin A levels were increased with persistent or recurrent (n = 9) stage III or IV ovarian cancer. Activin A levels dropped postoperatively and remained at or below the control level in patients in remission. CONCLUSION: Serum activin A levels correlate with recurrent or persistent disease in patients with epithelial ovarian cancer. PMID- 10385558 TI - Allelic loss on chromosome arm 8p: analysis of sporadic epithelial ovarian tumors. AB - OBJECTIVE: Our objective was to determine the frequency of allelic loss at 8p21 in sporadic epithelial ovarian cancer. We recently described allelic loss at this locus in 7/9 ovarian cancers from patients with BRCA1 gene mutations. METHODS: We anonymously obtained and examined 40 unselected invasive epithelial ovarian cancers and 5 low-malignant-potential (LMP) ovarian tumors for loss of heterozygosity (LOH) at 8p12-22. Pure epithelial and stromal cell populations were procured selectively by laser capture microdissection and extracted DNA was amplified with polymorphic microsatellite markers spanning the region of interest. RESULTS: LOH was highest (50%) at marker D8S136 located at 8p21 with 15 of 30 informative cases exhibiting an allelic deletion. None of the LMP tumors evaluated showed LOH at 8p12-22. A trend toward more frequent LOH at 8p12-22 was identified with increasing disease aggressiveness from LMP to early stage invasive ovarian cancer to advanced stage invasive ovarian cancer (Lehman's test, P2 < 0.024). CONCLUSIONS: Fifty percent allelic loss at the distal portion of 8p21 has not been reported to date for sporadic epithelial ovarian carcinomas. The higher rate of loss in our cohort, in contrast to previous allelotyping studies, is due likely to analysis from homogenous cell populations. These results, in concert with our previous study of BRCA1 mutation-positive patients, suggest a tumor suppressor gene locus at 8p21 for epithelial ovarian cancer. PMID- 10385559 TI - Epidemiologic differences between women with borderline ovarian tumors and women with epithelial ovarian cancer. AB - OBJECTIVE: The aim of this study was to study the relationship between borderline ovarian tumors (BLOT) and epithelial ovarian cancer (EOC) by comparing the epidemiologic features of women with BLOT with those of women with EOC of similar histology. MATERIAL AND METHODS: The epidemiologic features of 32 women with serous and mucinous BLOT were compared with those of 273 women with primary serous or mucinous EOC. We included all women with the documented respective histologic diagnoses admitted to Roswell Park Cancer Institute between 1982 and 1996 who returned a self-administered epidemiologic questionnaire which contained 44 items pertaining to reproductive, contraceptive, medical, social, dietary, occupational, and family histories of cancer. Individual variables between both groups were compared using the Student t test, chi2 analysis, the Mantel-Haenszel test, and the Wilcoxon nonparametric test. Two-tailed P < 0.05 was considered significant. RESULTS: The response rate to the questionnaire was 63% in the BLOT group and 60% in the EOC group. There was no significant difference between the two groups in geographic location, race, education, income, smoking, marital status, age at first pregnancy, age at first birth, history of hysterectomy, history of infertility, history of tubal surgery, use of hormone replacement therapy, or history of diaphragm or intrauterine contraceptive device use. There were no significant differences in family history of malignancy between women with BLOT and those with EOC. Women with BLOT were significantly younger than those with EOC (mean age 47 +/- 14.0 versus 56 +/- 13.7, P < 0.01). There was an apparent difference in oral contraceptive pill use between both groups. However, when we adjusted for age by stratification this difference was not significant (P = 0.089). CONCLUSIONS: The epidemiologic features of women with BLOT are similar to those of women with EOC with the exception of an earlier age of onset. These findings might be consistent with one etiology for both conditions. PMID- 10385561 TI - Pelvic abscess with fistula to the abdominal wall due to verrucous carcinoma. AB - The case report of a 38-year-old woman with a pelvic abscess resulting from verrucous carcinoma of the uterine cervix is presented. This case is remarkable because the abscess formed a fistula through the anterior abdominal wall and because there was no visible lesion on the cervix. The patient underwent a total abdominal hysterectomy, left salpingectomy, fistulectomy, and removal of the abscess. Diagnosis was made on pathologic examination of the extirpated specimen. Genital tract verrucous carcinoma and genitocutaneous fistulae are reviewed. PMID- 10385560 TI - Survival probability in ovarian clear cell adenocarcinoma. AB - OBJECTIVE: The aim of this study was to evaluate the 5-year survival probability (SP) of patients treated for ovarian clear cell adenocarcinoma (OCCA) at a single tertiary institution and to compare it to the 5-year SP of patients with other histologic subtypes of epithelial ovarian cancer. METHODS: Sixty-four patients with pure OCCA treated at the Cleveland Clinic Foundation from 1981 to 1996 were retrospectively identified and clinical information was abstracted. All histologic materials were reviewed by a single gynecologic pathologist. SP was calculated by the Kaplan-Meier method. SPs for OCCA patients were compared to that of other high-grade epithelial ovarian cancer patients in the gynecologic tumor registry. Cox proportional hazards modeling was used to identify varibles associated with decreased SP. RESULTS: The FIGO stages of OCCA study patients were Stage I, 31 (50%), Stage II, 6 (10%), Stage III, 17 (27%), and Stage IV, 8 (13%) (2 patients unstaged). Forty-four patients had no gross residual cancer at the completion of initial surgery while 9 patients had 1 cm residual. Forty-five (73%) received postoperative chemotherapy. The median follow-up for surviving patients is 97 months (range 38 to 209 months). The overall 5-year SP of OCCA patients is 50% with limited disease (Stages I and II) patients having a 5-year SP of 72% versus 17% 5-year SP in patients with advanced disease (P < 0.001). FIGO stage was most predictive of outcome. The overall 5-year SP of OCCA patients (50%) differed significantly (P < 0.05) from that of other ovarian cancer registry patients (30%). OCCA patients with limited cancer survived similarly to registry patients (72 vs 72%) as did patients with advanced OCCA compared with registry patients (17 vs 22%). CONCLUSIONS: When controlled for grade and stage, the overall survival with OCCA is identical to that of other high-grade epithelial ovarian cancers. Factors that account for the better overall survival of OCCA patients are more favorable disease stage, younger age, and improved debulking status. PMID- 10385562 TI - Endometrial carcinoma associated with pregnancy: A report of three cases and review of the literature. AB - Endometrial carcinoma associated with pregnancy is uncommon. In case 1, a 40-year old gravida 2, para 2, was diagnosed with focal well-differentiated papillary adenocarcinoma 4 months postpartum. In case 2, a 35-year-old gravida 1, para 0, was diagnosed with a well-differentiated papillary adenocarcinoma of the endometrium after a D&C for an incomplete abortion at 7 weeks gestation. In case 3, a 32-year-old gravida 2, para 1, was diagnosed with a moderately differentiated adenocarcinoma with squamous metaplasia 4 months postpartum. All are without evidence of disease more than 2 years after therapy. A literature review shows 24 previous cases of pregnancy associated with endometrial cancer. These cases demonstrate the importance of endometrial sampling for abnormal postpartum bleeding despite the protective effects of pregnancy. PMID- 10385564 TI - Contralateral pelvic and aortic lymph node metastasis in clinical stage I epithelial ovarian cancer. AB - Bilateral pelvic and aortic node lymphadenectomy is recommended for clinically localized unilateral epithelial ovarian adenocarcinoma (International Federation of Gynecologists and Obstetricians stage IA). The laterality of nodal metastasis in clinical stage I disease is rarely documented in the literature. Some authors have reported that ipsilateral node dissection is adequate for staging. A patient with contralateral pelvic and aortic lymph node metastasis and clinical stage I epithelial ovarian adenocarcinoma is presented. Pathologic findings were consistent with contralateral-only lymph node metastasis. This case illustrates the importance of bilateral lymph node sampling for appropriate staging of clinically localized epithelial ovarian cancer. PMID- 10385563 TI - Extraovarian granulosa cell tumor. AB - OBJECTIVE: The aim of this study was to report a case of extraovarian granulosa cell tumor and to describe its relevance to the histologic origin of granulosa cell tumors and to clinical practice. METHODS: The clinical course and histopathology of the case were reviewed, and a literature search for other reported cases was performed. RESULTS: A 67-year-old woman presented with postmenopausal bleeding and a pelvic mass. Laparotomy revealed a 16-cm mass arising from the right pelvic sidewall, filling the pelvis, and involving the bladder and rectosigmoid colon. Both ovaries appeared normal and were separate from the mass. Pathologic examination revealed granulosa cell tumor. A literature search revealed no recently reported cases of extraovarian granulosa cell tumor. CONCLUSIONS: Granulosa cell tumors can arise in locations other than the ovary and may be derived from the mesenchyme of the genital ridge. Women who have undergone oophorectomy may have the potential to develop granulosa cell tumors. PMID- 10385565 TI - Port-site metastasis following laparoscopic lymphadenectomy for adenosquamous carcinoma of the cervix. AB - Although incisional metastases following surgery for cervical cancer are extremely rare, port-site disease following minimal-access surgery is becoming increasingly reported. We report a case of a metastasis which occurred at a port site following laparoscopic removal of lymph nodes affected by cervical adenosquamous carcinoma. This report adds to the literature suggesting that cutaneous tumor deposition may be enhanced by this method of surgery. PMID- 10385566 TI - Well-differentiated mucinous carcinoma of the ovary and a coexisting Brenner tumor both exhibit amplification of 12q14-21 by comparative genomic hybridization. AB - Although the coexistence of mucinous ovarian neoplasms and Brenner tumors is well established, the histogenesis and developmental relationship between the two remain unknown. We used comparative genomic hybridization to analyze two such tumors occurring simultaneously, one in each ovary, in a patient. Amplification of 12q14-21 sequences was found in both tumors; in addition, both tumors also had other, different changes, four identified in the Brenner tumor and six in the mucinous carcinoma. The occurrence of the same genetic alteration in both tumors in this woman suggests that the mucinous carcinoma and Brenner tumor may be clonally related, i.e., one arose from the other by means of metastatic spreading of transformed cells from one ovary to the other. An alternative explanation is that some unknown, putative tumorigenic agent induced similar and synchronous pathogenetic changes in the epithelium of both ovaries. The phenotypic differences between the tumors are presumably attributable to the other unique genetic abnormalities identified in both tumor types. PMID- 10385567 TI - A case of clitoral metastasis without skin involvement from rectal cancer. AB - An 84-year-old woman visited a private dermatologist and gynecologist due to pain in the external genitals. However, no abnormality was found. She was referred to a surgeon in our hospital to clarify the etiology of the pain. Rectal cancer and liver metastatic tumor were detected, and the rectal cancer was resected. However, the pain increased after the operation and she was referred to our department. No macroscopic abnormalities of the external genitals were found. However, a vaginal examination could not be performed due to severe pain. By local examination under anesthesia, enlargement of the clitoris was detected. A simple clitoridectomy was performed. Histological examination revealed that the clitoral tumor was metastatic cancer originating from rectal cancer. PMID- 10385568 TI - Life-threatening tracheal metastasis complicating ovarian cancer--a case report. AB - BACKGROUND: Tracheal metastasis is a rare manifestation of recurrent ovarian cancer. CASE: We describe tracheal metastasis causing increasing respiratory distress in a patient with progressive stage IIIc undifferentiated serous papillary cancer involving the peritoneum and pleura and the retroperitoneal, diaphragmatic, parahilar, mediastinal, pretracheal, paratracheal, and supraclavicular lymph nodes. The situation necessitated rapid endoscopic laser ablation. CONCLUSION: Malignant tracheal obstruction should be considered in the differential diagnosis of patients with advanced ovarian cancer and respiratory distress. PMID- 10385569 TI - Relapse of acute lymphoblastic leukemia in pregnancy: survival following chemoirradiation and autologous transfer of interleukin-2-activated stem cells. AB - Four cases of relapse of acute lymphoblastic leukemia (ALL) in pregnancy have been reported previously. During the past 2 decades, ALL has become curable in a majority of children, many of whom have entered their reproductive years. Thus, additional occurrences of relapsing ALL during pregnancy can be anticipated. We present the fifth case in the English-language medical literature of recurrent ALL in pregnancy. A 20-year-old woman with ALL experienced a relapse during the third trimester of her first pregnancy. Reinduction therapy was started with vincristine and prednisone and the baby was delivered 3 weeks later. Umbilical cord blood was collected and stored. The patient then received intensive chemotherapy with whole body radiotherapy and autologous peripheral blood stem cell rescue. The ALL has been in second remission for 22 months. Our patient is the only current survivor of a relapse of ALL during pregnancy. In addition, the collection of umbilical cord blood from a pregnant woman with leukemia has not been reported previously. PMID- 10385570 TI - Do human normal somatic cells lack telomerase activity? PMID- 10385572 TI - Reply PMID- 10385571 TI - Correlation between in vitro drug response in the EDR assay and response to primary paclitaxel and cisplatin. PMID- 10385573 TI - Serum concentrations of endostatin in patients with vulvar cancer. PMID- 10385574 TI - Letter PMID- 10385575 TI - Letter PMID- 10385576 TI - How many subjects constitute a study? AB - In fMRI there are two classes of inference: one aims to make a comment about the "typical" characteristics of a population, and the other about "average" characteristics. The first pertains to studies of normal subjects that try to identify some qualitative aspect of normal functional anatomy. The second class necessarily applies to clinical neuroscience studies that want to make an inference about quantitative differences of a regionally specific nature. The first class of inferences is adequately serviced by conjunction analyses and fixed-effects models with relatively small numbers of subjects. The second requires random-effect analyses and larger cohorts. PMID- 10385578 TI - Functional specialization for semantic and phonological processing in the left inferior prefrontal cortex. AB - Neuroimaging and neuropsychological studies have implicated left inferior prefrontal cortex (LIPC) in both semantic and phonological processing. In this study, functional magnetic resonance imaging was used to examine whether separate LIPC regions participate in each of these types of processing. Performance of a semantic decision task resulted in extensive LIPC activation compared to a perceptual control task. Phonological processing of words and pseudowords in a syllable-counting task resulted in activation of the dorsal aspect of the left inferior frontal gyrus near the inferior frontal sulcus (BA 44/45) compared to a perceptual control task, with greater activation for nonwords compared to words. In a direct comparison of semantic and phonological tasks, semantic processing preferentially activated the ventral aspect of the left inferior frontal gyrus (BA 47/45). A review of the literature demonstrated a similar distinction between left prefrontal regions involved in semantic processing and phonological/lexical processing. The results suggest that a distinct region in the left inferior frontal cortex is involved in semantic processing, whereas other regions may subserve phonological processes engaged during both semantic and phonological tasks. PMID- 10385577 TI - The effect of normal aging on the coupling of neural activity to the bold hemodynamic response. AB - The use of functional neuroimaging to test hypotheses regarding age-related changes in the neural substrates of cognitive processes relies on assumptions regarding the coupling of neural activity to neuroimaging signal. Differences in neuroimaging signal response between young and elderly subjects can be mapped directly to differences in neural response only if such coupling does not change with age. Here we examined spatial and temporal characteristics of the BOLD fMRI hemodynamic response in primary sensorimotor cortex in young and elderly subjects during the performance of a simple reaction time task. We found that 75% of elderly subjects (n = 20) exhibited a detectable voxel-wise relationship with the behavioral paradigm in this region as compared to 100% young subjects (n = 32). The median number of suprathreshold voxels in the young subjects was greater than four times that of the elderly subjects. Young subjects had a slightly greater signal:noise per voxel than the elderly subjects that was attributed to a greater level of noise per voxel in the elderly subjects. The evidence did not support the idea that the greater head motion observed in the elderly was the cause of this greater voxel-wise noise. There were no significant differences between groups in either the shape of the hemodynamic response or in its the within-group variability, although the former evidenced a near significant trend. The overall finding that some aspects of the hemodynamic coupling between neural activity and BOLD fMRI signal change with age cautions against simple interpretations of the results of imaging studies that compare young and elderly subjects. PMID- 10385579 TI - The critical relationship between the timing of stimulus presentation and data acquisition in blocked designs with fMRI. AB - This paper concerns the experimental design and statistical models employed by fMRI activation studies which block presentation of linguistic stimuli. In particular, we note that the relationship between the timing of stimulus presentation and data acquisition can have a substantial impact on the ability to detect activations in critical language areas, even when the stimuli are presented in blocks. Using a blocked word rhyming paradigm and repeated investigations on a single subject, activation was observed in Broca's area (left inferior frontal cortex) and Wernicke's area (left posterior temporoparietal cortex) when (i) the timing of data acquisition was distributed throughout the peristimulus time and (ii) an event-related analysis was used to model the phasic nature of the hemodynamic response within each block of repeated word stimuli. In contrast, when the timing of data acquisition relative to stimulus presentation was fixed, activation was detected in Broca's area but not consistently in Wernicke's area. Our results indicate that phasic responses to stimuli occur even in a blocked design and that the sampling and proper modeling of these responses can have profound effects on their detection. Specifically, distributed sampling over peristimulus time is essential in order to detect small activations particularly when they are transient. These findings are likely to generalize to the detection of transient signals in any cognitive paradigm. PMID- 10385580 TI - Effective auditory-verbal encoding activates the left prefrontal and the medial temporal lobes: A generalization to illiterate subjects. AB - Recent event-related FMRI studies indicate that the prefrontal (PFC) and the medial temporal lobe (MTL) regions are more active during effective encoding than during ineffective encoding. The within-subject design and the use of well educated young college students in these studies makes it important to replicate these results in other study populations. In this PET study, we used an auditory word-pair association cued-recall paradigm and investigated a group of healthy upper middle-aged/older illiterate women. We observed a positive correlation between cued-recall success and the regional cerebral blood flow of the left inferior PFC (BA 47) and the MTLs. Specifically, we used the cued-recall success as a covariate in a general linear model and the results confirmed that the left inferior PFC and the MTL are more active during effective encoding than during ineffective encoding. These effects were observed during encoding of both semantically and phonologically related word pairs, indicating that these effects are robust in the studied population, that is, reproducible within group. These results generalize the results of Brewer et al. (1998, Science 281, 1185-1187) and Wagner et al. (1998, Science 281, 1188-1191) to an upper middle aged/older illiterate population. In addition, the present study indicates that effective relational encoding correlates positively with the activity of the anterior medial temporal lobe regions. PMID- 10385581 TI - The sensory somatotopic map of the human hand demonstrated at 4 Tesla. AB - Recent attempts at high-resolution sensory-stimulated fMRI performed at 1.5 T have had very limited success at demonstrating a somatotopic organization for individual digits. Our purpose was to determine if functional MRI at 4 T can demonstrate the sensory somatotopic map of the human hand. Sensory functional MRI was performed at 4 T in five normal volunteers using a low-frequency vibratory stimulus on the pad of each finger of the left hand. A simple motor control task was also performed. The data were normalized to a standard atlas, and individual and group statistical parametric maps (SPMs) were computed for each task. Volume of activation and distribution of cluster maxima were compared for each task. For three of the subjects, the SPMs demonstrated a somatotopic organization of the sensory cortex. The group SPMs demonstrated a clear somatotopic organization of the sensory cortex. The thumb to fifth finger were organized, in general, with a lateral to medial, inferior to superior, and anterior to posterior relationship. There was overlap in the individual SPMs between fingers. The sensory activation spanned a space of 12-18 mm (thumb to fifth finger) on the primary sensory cortex. The motor activation occurred consistently at the superior-most extent of the sensory activation within and across subjects. The sensory somatotopic map of the human hand can be identified at 4 T. High-resolution imaging at 4 T can be useful for detailed functional imaging studies. PMID- 10385582 TI - Areas 3a, 3b, and 1 of human primary somatosensory cortex. AB - This study defines cytoarchitectonic areas 3a, 3b, and 1 of the human primary somatosensory cortex by objective delineation of cytoarchitectonic borders and ensuing cytoarchitectonic classification. This avoids subjective evaluation of microstructural differences which has so far been the only way to structurally define cortical areas. Ten brains were fixed in formalin or Bodian's fixative, embedded in paraffin, sectioned as a whole in the coronal plane at 20 microm, and cell stained. Cell bodies were segmented from the background by adaptive thresholding. Equidistant density profiles (125 microm wide, spacing 300 or 150 microm) were extracted perpendicularly to the pial surface across cortical layers II-VI and processed with multivariate statistical procedures. Positions of significant differences in shape between adjacent groups of profiles were correlated with the cytoarchitectonic pattern. Statistically significant borders can be reproduced at corresponding positions across a series of nearby sections. They match visible changes in cytoarchitecture in the cell-stained sections. Area 3a lies in the fundus of the central sulcus, and area 3b in the rostral bank of the postcentral gyrus. Area 1 lies on its crown and reaches down into the postcentral sulcus. Interareal borders, however, do not match macrostructural landmarks of the postcentral gyrus, and they considerably vary in their positions relative to these landmarks across different brains. Hence, only genuine microstructural analysis can define the borders between these cortical areas. Additional significant borders which do not correlate with visible changes in cytoarchitecture can be found within areas 3b and 1. They may represent somatotopy and/or cortical representations of different somatosensory receptors. PMID- 10385583 TI - Cortical activation during perception of a rotating wide-field acoustic stimulus. AB - We describe sound stimuli that produce the perception of complete rotation around the head. Such stimuli are analogous to wide-field motion stimuli used in visual research, though auditory stimuli, unlike visual stimuli, can be perceived at any point around the head; they are the only cues for spatial perception behind the subject. Using PET on six subjects, we have compared regional brain activity during the perception of such motion stimuli, with the perception of a control stimulus producing equivalent amplitude changes without rotation. Rotation produced activation of the premotor cortex bilaterally and the right superior parietal cortex. The premotor activation involved the frontal eye fields and ventral premotor areas. The bifrontal and right parietal activation is consistent with previous demonstrations of activation within a frontoparietal network of areas during perception of a linear motion stimulus. The inferior premotor activation in this experiment may reflect preparation for head turning in response to auditory targets that cannot be tracked visually. PMID- 10385584 TI - Notice to subscribers and contributors PMID- 10385585 TI - Immunotherapy with antigens and epitopes: pro or con. PMID- 10385586 TI - Nucleosides and nucleotides in the lung: role in asthma. PMID- 10385587 TI - Immune therapy for lung cancer: are we getting closer? PMID- 10385588 TI - CD10/neutral endopeptidase inhibition augments pulmonary neuroendocrine cell hyperplasia in hamsters treated with diethylnitrosamine and hyperoxia. AB - In previous studies, we demonstrated that pulmonary neuroendocrine cell (PNEC) hyperplasia in hamsters treated with diethylnitrosamine (DEN) plus 65% hyperoxia (DEN/O2) reflects predominantly neuroendocrine cell differentiation. Several peptides implicated in non-neoplastic PNEC hyperplasia are hydrolyzed by CD10/neutral endopeptidase 24.11 (CD10/NEP), an enzyme known to downregulate neurogenic inflammation of the lung by modulating locally effective concentrations of multiple bioactive peptides. In fetal mice, we observed that CD10/NEP inhibition by SCH32615 potentiates cell proliferation and type II cell differentiation in the lung in utero. Further, CD10/NEP messenger RNA levels parallelled relative PNEC numbers in DEN/O2-treated hamster lung, suggesting that the enzyme might mediate spontaneous regression of PNEC hyperplasia. The goals of the present study were: (1) to determine whether CD10/NEP inhibition would alter the extent of PNEC hyperplasia occurring in these hamsters, and (2) to analyze cellular mechanisms potentially involved in altering numbers of PNECs in this model. We administered SCH32615 chronically to a subset of DEN/O2-treated hamsters. Immunostaining of lungs from the CD10/ NEP-inhibited subset demonstrated significant acceleration of the development of PNEC hyperplasia, increased PNEC proliferation, and diminished PNEC apoptosis as compared with animals receiving no SCH32615. These observations indicate that PNEC hyperplasia can occur as a result of multiple cellular processes, including increased neuroendocrine cell differentiation, proliferation, and survival. CD10/NEP modulates PNEC numbers primarily by promoting cell differentiation and proliferation during lung injury, probably via increasing the half-life of bioactive peptides in the lung. PMID- 10385589 TI - Opposite effects of immunotherapy with ovalbumin and the immunodominant T-cell epitope on airway eosinophilia and hyperresponsiveness in a murine model of allergic asthma. AB - In the present study, we investigated immunotherapy using an entire protein or an immunodominant epitope in a murine model of allergic asthma. Immunotherapy was performed in ovalbumin (OVA)-sensitized mice before OVA challenge. Mice were treated subcutaneously with OVA, the immunodominant epitope OVA323-339, or vehicle. In vehicle-treated animals, repeated OVA challenge induced increased serum levels of OVA-specific immunoglobulin (Ig)G1, IgE, airway eosinophilia, and hyperresponsiveness, compared with saline-challenged animals. In addition, interleukin (IL)-4 and IL-5 production upon OVA restimulation of lung-draining lymph node cells in vitro were significantly increased in OVA-challenged animals. Immunotherapy using OVA significantly reduced airway eosinophilia and hyperresponsiveness. This finding was accompanied by significantly reduced OVA specific IL-4 and IL-5 production. Further, OVA immunotherapy induced increased serum levels of OVA-specific IgG1, whereas OVA-specific IgG2a and IgE levels were not affected. In contrast to OVA immunotherapy, immunotherapy with OVA323-339 aggravated airway eosinophilia and hyperresponsiveness. OVA-specific IgG1, IgG2a, and IgE serum levels, and in vitro IL-4 and IL-5 production, were not affected. Thus, immunotherapy with protein resulted in beneficial effects on airway eosinophilia and hyperresponsiveness, which coincided with a local reduced T helper 2 (Th2) response. In contrast, peptide immunotherapy aggravated airway hyperresponsiveness and eosinophilia, indicating a local enhanced Th2 response. PMID- 10385590 TI - Mechanisms of the potentiation by adenosine of adenosine triphosphate-induced calcium release in tracheal smooth-muscle cells. AB - The effects of concomitant P1-receptor stimulation on peak intracellular Ca2+ release by extracellular adenosine 5'-triphosphate (ATP) and 5-hydroxytryptamine (5-HT) were investigated in cultured airway smooth-muscle (ASM) cells. The results show that peak Ca2+ release to ATP is enhanced by preincubation with adenosine (ADO) and with the specific A3 receptor agonist 1-Deoxy-1-(6-([(3 iodophenyl)methyl] amino)-9H-purin-9-yl)-N-methyl-beta-D-ribofuranuronamide (1B MECA). The response to 5-HT, a smooth-muscle contractile agonist, was also enhanced after preincubation with ADO. Further measurements showed that this enhancement of the response to ATP was dependent on extracellular calcium because it was abolished by the removal of Ca2+ from the extracellular fluid and by incubation with the calcium channel blocker nifedipine. In addition, there was no difference between the levels of total inositol phosphates measured in the presence of ATP alone or of ADO + ATP. AACOCF3, a specific blocker of phospholipase A2, decreased the peak Ca2+ response to ATP and abolished the enhanced response to ATP and 5-HT produced by ADO. We conclude that stimulation of P1 and P2 receptors in ASM cells activates not only phospholipase C but also phospholipase A2. The enhancement of ATP-induced and 5-HT-induced Ca2+ release is due to Ca2+ influx from the extracellular fluid through a Ca2+ channel presumably modulated by arachidonic acid. These data show that endogenous ADO may modulate airway hyperresponsiveness by enhancing the ASM response to contractile agonists. PMID- 10385591 TI - DNA vaccination against HuD antigen elicits antitumor activity in a small-cell lung cancer murine model. AB - There is a clinically significant correlation between the presence of an antibody against the paraneoplastic encephalomyelitis antigen HuD and the limitation of tumor spread in patients with small-cell lung cancer (SCLC). This suggests that HuD is a possible target molecule for antitumor immunotherapy against SCLC. We have hypothesized that anti-HuD immunity suppresses in vivo growth of HuD expressing tumor cells. In this study, Colon 26, a murine adenocarcinoma cell line, stably transfected with the HuD gene (Colon 26/HuD cell) was used as a target cell, and the immunity against HuD was evoked by intramuscular injection of a HuD-expressing plasmid, a technique of DNA vaccination previously used in BALB/c mice. Colon 26/HuD cells were injected subcutaneously and tumor size was calculated as a product of width and length. Antitumor activity was investigated by using two different lots of Colon26/HuD cells in two protocols: Protocol 1, in which either Colon 26/HuD or Colon 26 cells were injected in each side, and Protocol 2, in which Colon 26/HuD cells alone were injected. The size of Colon 26/HuD tumors obtained from mice vaccinated with HuD-expressing plasmid was significantly smaller than those from negative control plasmid-vaccinated mice (86.6 +/- 29.9 versus 195.3 +/- 48.1 mm2, P < 0.05 in Protocol 1; 107.7 +/- 12.8 versus 156.6 +/- 22.8 mm2, P < 0.05 in Protocol 2). Moreover, the de novo DNA synthesis of spleen cells obtained from HuD-vaccinated mice was significantly enhanced. In addition, anti-HuD antibody was found in individual sera obtained from HuD-vaccinated mice. DNA vaccination with mouse HuD antigen suppressed HuD expressing tumor growth in a murine SCLC model. PMID- 10385592 TI - Early events in naphthalene-induced acute Clara cell toxicity: comparison of membrane permeability and ultrastructure. AB - Naphthalene causes severe dose- and site-selective injury to mouse nonciliated bronchiolar (Clara) epithelial cells. Toxicity is characterized by exfoliation of injured Clara cells into the airway lumen 24 h after exposure. The purpose of this study was to define the temporal pattern of intracellular changes immediately following naphthalene treatment, with the goal of identifying critical early events involved in cytotoxicity. Mice were injected with naphthalene or carrier and were killed 1, 2, 3, and 6 h after treatment (PT). Loss of membrane integrity was assessed by ethidium homodimer-1 permeability and confocal microscopy. Cell morphology and ultrastructure were evaluated using high resolution light and electron microscopy. Permeable cells were found only in terminal bronchioles and increased in abundance with time PT. At 2 and 3 h PT, when most Clara cells had early signs of injury, few permeable cells were detected. Many Clara cells had apical membrane blebs that contained abundant, swollen, smooth endoplasmic reticulum (SER) and few other organelles. By 6 h PT many Clara cells were membrane-permeable. However, many permeable Clara cells lacked apical blebs and SER was less abundant in these cells. Cytoplasmic blebbing may be a mechanism to protect the cell by isolating and removing damaged SER. We conclude that the early stages of injury include SER swelling and bleb formation which precede increases in cell membrane permeability after acute naphthalene injury to bronchiolar Clara cells in vivo. PMID- 10385593 TI - Thiol depletion induces apoptosis in cultured lung fibroblasts. AB - Thiol antioxidants are implicated in the protection of cells from oxidative injury. We studied the role of thiols in the regulation of apoptosis in cultured lung fibroblasts. Thiol depletion by culturing fibroblasts in cystine-free medium or with thiol-depleting agents induced oxidant accumulation and cell death by apoptosis. The cell death was prevented by the antioxidants ascorbic acid (AA) and catalase. Thiol depletion also induced leukotriene (LT) C4, LTD4, and LTE4 production and selective phosphorylation of p38-mitogen-activated protein kinase (MAPK) and its nuclear substrate ATF2. LT production and p38-MAPK phosphorylation were required for induction of apoptosis because thiol depletion-induced apoptosis was completely blocked by the 5-lipoxygenase inhibitor AA861, the LT antagonists FPL55712 and ONO1078, and the p38-MAPK inhibitor SB203580. LT production was inhibited by AA and p38-MAPK phosphorylation was inhibited by AA, AA861, and FPL55712. In an in vitro scratch wound model, repopulating fibroblasts at the wound margin, but not quiescent cells at the intact site, selectively underwent thiol depletion- induced apoptosis that was completely blocked by AA861, FPL55712, and SB203580. Thus, thiol depletion induces apoptosis through an ordered pathway involving oxidant accumulation, LT production, and p38-MAPK activation. Apoptosis of wound fibroblasts may be responsible for impaired wound healing in various organs, including the lung. PMID- 10385594 TI - Lung growth response after tracheal occlusion in fetal rabbits is gestational age dependent. AB - In utero tracheal occlusion (TO) is a potent stimulus of fetal lung growth, and is currently being applied in clinical trials to treat severe forms of pulmonary hypoplasia. The aim of this study was to examine the effect of timing of TO on pulmonary growth and maturation rates. Fetal rabbits (term = 31 d) were subjected to in utero tracheal clipping at 24 (late pseudoglandular stage) or 27 d of gestation (late canalicular/early terminal sac stage). Sham-operated littermates served as controls (C). Animals were killed at time intervals ranging from 1 to 6 d (early group) or 1 to 3 d (late group) after occlusion. Lung growth was measured by computerized stereologic volumetry and 5'-bromo-2'-deoxyuridine (BrdU) pulse labeling. Pneumocyte II population kinetics were analyzed using a combination of anti-surfactant protein-A and BrdU immunohistochemistry and computer-assisted morphometry. Statistical analysis was performed using unpaired Student's t test. Early TO was followed by an initial 3-d stagnation of growth and subsequently a dramatic acceleration of growth (BrdU-labeling index [LI] 10.1 +/- 0. 6% in TO versus 2.7 +/- 0.5% in C at 29 d, P < 0.001). In contrast, late TO induced an immediate and sustained moderate increase of lung growth (BrdU-LI 2.8 +/- 0.9% in TO versus 1.1 +/- 0.2% in C at 30 d, P < 0.05), associated with relatively more pronounced air-space distension. Whereas late TO caused no significant alterations in type II cell density or proliferation, early TO was followed by a marked increase in type II cell proliferation, paradoxically associated with dramatic reduction of type II cell density after 29 d. The effects of intrauterine TO on fetal lung growth and type II cell kinetics critically depend on the gestational age, and thus on the maturity of the lungs at the time of surgery. These findings have important clinical implications with respect to the timing of fetal interventions aimed at promoting lung growth. The fetal rabbit provides an invaluable model to study the mechanics and age dependency of TO-induced lung growth. PMID- 10385595 TI - Glucocorticoids inhibit proliferation, cyclin D1 expression, and retinoblastoma protein phosphorylation, but not activity of the extracellular-regulated kinases in human cultured airway smooth muscle. AB - We have previously shown that glucocorticoids inhibit mitogen-stimulated proliferation of human cultured airway smooth muscle (ASM) cells. The present study analyzed the effect of glucocorticoids on key regulatory pathways leading to passage of cells through the restriction point of the cell cycle, including those mediated by extracellular-regulated kinases (ERK) 1 and 2; the ERK upstream regulator MAPK kinase (MEK1); cyclin D1 levels; and levels and phosphorylation of retinoblastoma protein (pRb). Fluticasone propionate, a new inhaled glucocorticoid, was at least 10-fold more potent than dexamethasone in inhibiting thrombin-stimulated DNA synthesis and increases in cell number. Thrombin stimulated increases in the levels and hyperphosphorylation of pRb were inhibited by glucocorticoids, which also reduced thrombin-stimulated cyclin D1 protein and messenger RNA (mRNA) levels. PD98059 (10 microM), an inhibitor of MEK1 activation, markedly attenuated thrombin stimulation of ERK activity and phosphorylation, DNA synthesis, and cyclin D1 levels. However, glucocorticoids had no effect on ERK activity or phosphorylation at 5 min, 2 h, or 12 h after addition of thrombin. In conclusion, glucocorticoid-induced reduction of cyclin D1 mRNA and protein levels, and of pRb phosphorylation, is sufficient to account for inhibition of ASM proliferation. Furthermore, these inhibitory effects of glucocorticoids on cyclin D1 and pRb occur on a component of the mitogen signaling cascade that is either downstream of or parallel to the ERK pathway. PMID- 10385596 TI - Mild vitamin A deficiency delays fetal lung maturation in the rat. AB - During late pregnancy, the fetal lung stores surfactant in preparation for extrauterine life. Surfactant deficiency, most often due to prematurity, precipitates respiratory distress syndrome (RDS) of the neonate. Although vitamin A (retinol) and retinoic acid have been shown to enhance the synthesis of phospholipid surfactant components, their effect on surfactant-specific proteins is unclear. No attempt has been made to evaluate the consequences of vitamin A restriction on surfactant phospholipid storage or on the expression of the life essential surfactant protein-B (SP-B). We induced in rats a partial vitamin A deficiency leading to a 30-60% reduction in blood retinol, a status compatible with maintenance of gestation and absence of gross abnormalities in offspring. At term, lung surfactant phospholipids were reduced by 21%, and the major surfactant phospholipid, disaturated phosphatidylcholine (DSPC), was reduced by 27% in vitamin A-deficient (VAD) fetuses. The decrease in surfactant phospholipids and DSPC correlated linearly with plasma retinol, and reached about 50% in fetuses with the lowest retinol concentrations; it was accompanied by reduced expression of the gene for fatty acid synthase, a key enzyme in the synthetic pathway for surfactant-phospholipid lipid precursors. The amounts of SP-A, SP-B, and SP-C messenger RNAs were decreased by 46%, 32%, and 28%, respectively, in VAD fetuses. Consistently, amounts of SP-A and SP-B proteins were diminished as assessed by Western blotting. The proportion of type II cells determined after SP-B labeling was unchanged in VAD as compared with control lungs. Vitamin A deficiency is therefore a cause of lung maturational delay. In view of its rather large incidence in human populations, it may represent an increased risk for RDS and an aggravating factor for prematurity. PMID- 10385597 TI - 5-Oxo-6,8,11,14-eicosatetraenoic acid induces important eosinophil transmigration through basement membrane components: comparison of normal and asthmatic eosinophils. AB - Basement membrane transmigration is an important step in tissue recruitment of eosinophils into inflamed tissue. Recent reports showed that this phenomenon is modulated by platelet-activating factor (PAF) in combination with cytokines and proteinases. We investigated the in vitro efficacy of 5-oxo-6,8,11, 14 eicosatetraenoic acid (5-oxo-ETE), a metabolite of arachidonic acid and known as a potent eosinophil chemotactic factor, in promoting the transmigration of blood eosinophils from normal and asthmatic subjects through a Matrigel basement membrane. 5-Oxo-ETE proved to be a more potent (> 10-fold) inducer of eosinophil transmigration than PAF, and this effect was similar in cells from normal and asthmatic subjects (82.0 +/- 3.7% and 88.1 +/- 3.7%, respectively). Moreover, 5 oxo-ETE was active in the absence of interleukin (IL)-5, although this cytokine amplified the effect of 5-oxo-ETE from 61.3 +/- 3.3% to 92.8 +/- 1.8% (p = 0.003). The membrane receptor for urokinase plasminogen activator (CD87), a serine protease, was observed on eosinophils, and its expression was increased by IL-5. The inhibition of both metalloproteinases (MMP) and plasmin/plasminogen complex with inhibitor or monoclonal antibodies decreased cell transmigration by about 50%. Combination of an MMP inhibitor with anti-CD87 antibodies had no additive effect. These data show that 5-oxo-ETE is an efficient promoter of eosinophil transmigration in vitro, and is much more potent in this respect than PAF. The data suggest that 5-oxo-ETE could play an important role in eosinophil recruitment in vivo. Moreover, they demonstrate that in addition to MMP, the plasmin/plasminogen system could be involved in eosinophil transmigration. PMID- 10385598 TI - Autocrine function of inducible nitric oxide synthase and cyclooxygenase-2 in proliferation of human and rat pulmonary artery smooth-muscle cells: species variation. AB - Pulmonary hypertension is characterized by hypertrophy and hyperplasia of vascular smooth muscle occurring via an unknown mechanism. Cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) are expressed under inflammatory conditions and produce mediators that regulate growth in some tissues. We have therefore addressed the question of COX-2 and iNOS involvement in proliferation of human and rat pulmonary artery (PA) smooth-muscle cells (SMC). Interleukin (IL)-1beta suppressed proliferation of both human and rat PA SMC. Moreover, IL 1beta induced COX-2 expression in both cell types. By contrast, IL-1beta stimulated the expression of iNOS protein in rat cells only. COX-2 induced in human cells inhibited proliferation, whereas COX-2 products in rat cells were without affect. However, iNOS activity in rat cells suppressed their proliferation. We conclude that human and rat evolution has diverged such that COX-2 and iNOS, although induced by the same mediator, have different levels of activity and functions in the two species. In humans, induction of COX-2 during pulmonary hypertension may be beneficial for long-term treatment of this disease. PMID- 10385599 TI - beta-hexosaminidase-induced activation of p44/42 mitogen-activated protein kinase is dependent on p21Ras and protein kinase C and mediates bovine airway smooth muscle proliferation. AB - Late-phase and sustained activation of p44/42(MAPK) has been reported to be a critical factor in cell mitogenesis. We therefore hypothesized that p44/42(MAPK) is involved in mannosyl-rich glycoprotein-induced mitogenesis in bovine airway smooth-muscle cells (ASMC). Treatment of adherent ASMC with beta-hexosaminidase A (Hex A, 50 nM), an endogenous mannosyl-rich glycoprotein, resulted in a late onset (30-min) activation of p44/42(MAPK) that lasted for 4 h. Activation of p44/42(MAPK) induced by Hex A was inhibited by an 18-mer phosphorothioate derivatized antisense oligonucleotide (1-5 microM) directed to human p44(MAPK); the mitogen-activated protein kinase kinase (MEK1) inhibitor PD98059 (5 microM); the p42(MAPK) inhibitor Tyrphostin AG-126 (0.2 microM); the farnesyl transferase inhibitors SCH-56582 (10 microM) and FPT III (10 miroM), which inhibit p21Ras activation; and Calphostin C (0.2 microM), an inhibitor of protein kinase C. These agents also inhibited Hex A-induced cell proliferation in bovine ASMC. These data suggest that Hex A activates p44/42(MAPK) in a p21Ras- and PKC dependent manner and that this activation mediates Hex A- induced mitogenesis in bovine ASMC. PMID- 10385600 TI - Hypoxic modulation of manganese superoxide dismutase promoter activity and gene expression in lung epithelial cells. AB - We investigated the effects of hypoxia (< 2.5% O2) on rat manganese superoxide dismutase (MnSOD) gene promoter-luciferase reporter constructs in transiently transfected lung epithelial cells (A549, L2, and E1A-T2) and fibroblasts (R9Ab). We cloned MnSOD promoter-luciferase reporter constructs (numbers refer to length in base pairs [bp] in the 5' direction from the transcription initiation site): 2,505, 1,064, 507, 405, and 289 into pGL2-Basic, a promoterless, firefly luciferase vector. Lung cells were transfected with MnSOD promoter-reporter constructs with or without thymidine kinase-driven Renilla luciferase (pRL-TK), and were exposed to air/5% CO2 or hypoxia (2.5% O2/5% CO2/balance N2) for 24 h. Hypoxia caused a significant (by two-way analysis of variance) consistent increase in luciferase in the A549 cell (human lung carcinoma) line. Greatest expression (> 3-fold increase) in hypoxia was associated with the 2,505-bp MnSOD promoter (normalized to cellular protein). Azide (10 microM) did not increase expression of the MnSOD reporter constructs. The 289-bp promoter was sufficient to express the reporter in air and to increase its expression in hypoxia. Promoter activity of the rat MnSOD 5' region, assessed by luciferase reporter constructs in A549 cells, increased in hypoxia. The increase was exclusive to A549 cells and did not occur in other cells. PMID- 10385602 TI - Soluble tumor necrosis factor (TNF) receptors p55 and p75 and interleukin-10 downregulate TNF-alpha activity during the lung response to silica particles in NMRI mice. AB - We have found reduced activity of tumor necrosis factor (TNF)-alpha accompanying resolving and fibrosing alveolitis induced in NMRI mice by mineral particles (MnO2 and SiO2, respectively), which is in apparent contradiction to the well recognized proinflammatory and profibrotic activities of this cytokine. The objective of this study was to examine the mechanisms involved in this paradoxical response in NMRI mice. Although lung tissue messenger RNA (mRNA) levels for TNF-alpha were transiently (up to 15 d) and persistently (up to 120 d) upregulated in the resolving and fibrosing models, respectively, these changes were not accompanied by a parallel release of TNF-alpha protein, which was respectively transiently and persistently downregulated in bronchoalveolar lavage fluid and bronchoalveolar lavage cell cultures. The downregulation of the TNF alpha protein was concurrent with the accumulation of recruited polymorphonuclear neutrophils (PMNs) in alveoli, and coculture experiments showed that PMN explanted from the lungs of mice treated with silica particles were able to downregulate the expression of TNF-alpha protein by naive alveolar macrophages. In addition, PMN depletion prevented the downregulation of TNF-alpha induced by silica, further establishing the role of PMNs in the downregulation of TNF-alpha. The possible degradation of TNF-alpha by proteolytic enzymes could be excluded. Marked increases in soluble p55 and p75 TNF receptors (sTNF-R), as well as in interleukin (IL)-10, paralleled the downregulation of TNF-alpha protein. The role of these mediators in the observed reduction of TNF-alpha activity was confirmed by immunoneutralizing the activity of p55 and p75 sTNF-R and by using IL-10 deficient animals. Because IL-10 also exerts profibrotic activity in addition to its antiinflammatory activity, the protracted overproduction of IL-10 observed in fibrosing alveolitis may help the understanding of why, in NMRI mice treated with silica particles, lung fibrosis develops in association with a downregulation of TNF-alpha. PMID- 10385601 TI - Reactive oxygen and nitrogen intermediates increase transforming growth factor beta1 release from human epithelial alveolar cells through two different mechanisms. AB - Transforming growth factor (TGF)-beta1 is a growth factor involved in the mechanisms of lung repair and fibrosis that follow inflammatory processes. We sought to examine the link between the generation of reactive oxygen intermediates (ROI) or reactive nitrogen intermediates (RNI) by inflammatory cells and the expression of TGF-beta1 by alveolar epithelial cells. Exposure of the A549 lung epithelial cell line to either an ROI generating system (xanthine and xanthine oxidase) or an RNI donor (S-nitroso-N-acetyl-penicillamine [SNAP]) promoted a time- and dose-dependent increase in TGF-beta1 release, as measured by a specific enzyme-linked immunosorbent assay. At the peak, the levels of TGF beta1 were twice the control values. The induction of TGF-beta1 release by ROI was blunted by catalase and unaffected by superoxide dismutase, indicating the involvement of hydrogen peroxide. The response was also blunted by 5, 6-dichloro 1-beta-D-ribofuranosyl benzimidazole (DRB), a specific RNA polymerase II inhibitor, and accompanied by a corresponding increase in TGF-beta1 messenger RNA, as measured by quantitative/competitive reverse transcription polymerase chain reaction, suggesting the involvement of transcriptional mechanisms and possibly other downstream mechanisms. In contrast, RNI-induced TGF-beta1 release was unaffected by DRB and blunted by the protein synthesis inhibitor cycloheximide, suggesting the involvement of translational and post-translational mechanisms. This response required cyclic guanosine monophosphate (cGMP)- mediated processes because (1) immunoreactive cGMP accumulated in the culture medium of SNAP-treated cells; (2) SNAP-induced TGF-beta1 release was blunted by KT 5823, an inhibitor of cGMP-dependent protein kinase; and (3) similar increase in TGF-beta1 release was obtained by cell exposure to membrane-permeable dibutyryl-cGMP or to atrial natriuretic factor, a known agonist of particulate guanylate cyclase. These data suggest that in vitro exposure of human alveolar epithelial cells to ROI and RNI enhances TGF-beta1 release through different mechanisms. In vivo, this control may constitute a molecular link between inflammatory and fibrotic processes. PMID- 10385603 TI - Mass spectrometric quantification of amino acid oxidation products in proteins: insights into pathways that promote LDL oxidation in the human artery wall. AB - Oxidatively damaged low density lipoprotein (LDL) may play an important role in atherogenesis, but the physiologically relevant pathways have proved difficult to identify. Mass spectrometric quantification of stable compounds that result from specific oxidation reactions represents a powerful approach for investigating such mechanisms. Analysis of protein oxidation products isolated from atherosclerotic lesions implicates tyrosyl radical, reactive nitrogen species, and hypochlorous acid in LDL oxidation in the human artery wall. These observations provide chemical evidence for the reaction pathways that promote LDL oxidation and lesion formation in vivo.--Heinecke, J. W. Mass spectrometric quantification of amino acid oxidation products in proteins: insights into pathways that promote LDL oxidation in the human artery wall. PMID- 10385604 TI - Effects of nitric oxide and nitric oxide-derived species on prostaglandin endoperoxide synthase and prostaglandin biosynthesis. AB - Prostaglandins and NO. are important mediators of inflammation and other physiological and pathophysiological processes. Continuous production of these molecules in chronic inflammatory conditions has been linked to development of autoimmune disorders, coronary artery disease, and cancer. There is mounting evidence for a biological relationship between prostanoid biosynthesis and NO. biosynthesis. Upon stimulation, many cells express high levels of nitric oxide synthase (NOS) and prostaglandin endoperoxide synthase (PGHS). There are reports of stimulation of prostaglandin biosynthesis in these cells by direct interaction between NO. and PGHS, but this is not universally observed. Clarification of the role of NO. in PGHS catalysis has been attempted by examining NO. interactions with purified PGHS, including binding to its heme prosthetic group, cysteines, and tyrosyl radicals. However, a clear picture of the mechanism of PGHS stimulation by NO. has not yet emerged. Available studies suggest that NO. may only be a precursor to the molecule that interacts with PGHS. Peroxynitrite (from O2.-+NO.) reacts directly with PGHS to activate prostaglandin synthesis. Furthermore, removal of O2.- from RAW 267.4 cells that produce NO. and PGHS inhibits prostaglandin biosynthesis to the same extent as NOS inhibitors. This interaction between reactive nitrogen species and PGHS may provide new approaches to the control of inflammation in acute and chronic settings. PMID- 10385605 TI - Is NF-kappaB the sensor of oxidative stress? AB - NF-kappaB is a dimeric transcription factor that is involved in the regulation of a large number of genes that control various aspects of the immune and inflammatory response. It is activated by a variety of stimuli ranging from cytokines, to various forms of radiation, to oxidative stress (such as exposure to H2O2). Recent studies have advanced our understanding of the signal transduction pathway leading to NF-kappaB activation by cytokines and will provide insights for the mechanism by which NF-kappaB is regulated by oxidative stress. An important question that is yet to be answered is whether reactive oxygen species play a physiological role in NF-kappaB activation. PMID- 10385606 TI - Vitamin E: function and metabolism. AB - Although vitamin E has been known as an essential nutrient for reproduction since 1922, we are far from understanding the mechanisms of its physiological functions. Vitamin E is the term for a group of tocopherols and tocotrienols, of which alpha-tocopherol has the highest biological activity. Due to the potent antioxidant properties of tocopherols, the impact of alpha-tocopherol in the prevention of chronic diseases believed to be associated with oxidative stress has often been studied, and beneficial effects have been demonstrated. Recent observations that the alpha-tocopherol transfer protein in the liver specifically sorts out RRR-alpha-tocopherol from all incoming tocopherols for incorporation into plasma lipoproteins, and that alpha-tocopherol has signaling functions in vascular smooth muscle cells that cannot be exerted by other forms of tocopherol with similar antioxidative properties, have raised interest in the roles of vitamin E beyond its antioxidative function. Also, gamma-tocopherol might have functions apart from being an antioxidant. It is a nucleophile able to trap electrophilic mutagens in lipophilic compartments and generates a metabolite that facilitates natriuresis. The metabolism of vitamin E is equally unclear. Excess alpha-tocopherol is converted into alpha-CEHC and excreted in the urine. Other tocopherols, like gamma- and delta-tocopherol, are almost quantitatively degraded and excreted in the urine as the corresponding CEHCs. All rac alpha-tocopherol compared to RRR-alpha-tocopherol is preferentially degraded to alpha-CEHC. Thus, there must be a specific, molecular role of RRR-alpha-tocopherol that is regulated by a system that sorts, distributes, and degrades the different forms of vitamin E, but has not yet been identified. In this article we try to summarize current knowledge on the function of vitamin E, with emphasis on its antioxidant vs. other properties, the preference of the organism for RRR-alpha tocopherol, and its metabolism to CEHCs. PMID- 10385608 TI - Regulation of hepatic glutathione synthesis: current concepts and controversies. AB - Glutathione (GSH) is an important intracellular peptide with multiple functions ranging from antioxidant defense to modulation of cell proliferation. GSH is synthesized in the cytosol of all mammalian cells in a tightly regulated manner. The major determinants of GSH synthesis are the availability of cysteine, the sulfur amino acid precursor, and the activity of the rate-limiting enzyme, gamma glutamylcysteine synthetase (GCS). In the liver, major factors that determine the availability of cysteine are diet, membrane transport activities of the three sulfur amino acids cysteine, cystine and methionine, and the conversion of methionine to cysteine via the trans-sulfuration pathway. Many conditions alter GSH level via changes in GCS activity and GCS gene expression. These include oxidative stress, activators of Phase II detoxifying enzymes, antioxidants, drug resistant tumor cell lines, hormones, cell proliferation, and diabetes mellitus. Since the molecular cloning of GCS, much has been learned about the regulation of this enzyme. Both transcriptional and post-transcriptional mechanisms modulate the activity of this critical cellular enzyme.--Lu, S. C. Regulation of hepatic glutathione synthesis: current concepts and controversies. PMID- 10385607 TI - New developments in the isoprostane pathway: identification of novel highly reactive gamma-ketoaldehydes (isolevuglandins) and characterization of their protein adducts. AB - The bicyclic endoperoxide prostaglandin (PG) H2 undergoes nonenzymatic rearrangement not only to PGE2 and PGD2, but also to levuglandins (LG) E2 and D2, which are highly reactive gamma-ketoaldehydes. Isoprostanes (IsoPs) are PG-like compounds that are produced by nonenzymatic peroxidation of arachidonic acid. PGH2-like endoperoxides are intermediates in this pathway. Therefore, we explored whether the IsoP endoperoxides also undergo rearrangement to form IsoLGs. Oxidation of arachidonic acid in vitro resulted in the formation of abundant quantities of compounds that were established to be IsoLGs by using mass spectrometric analyses. However, the formation of IsoLGs could not be detected in biological systems subjected to an oxidant stress. We hypothesized that this was due to extremely rapid adduction of IsoLGs to proteins. This notion was supported by the finding that LGE2 adducted to albumin at a rate that exceeded that of 4 hydroxynonenal by several orders of magnitude: >50% of LGE2 had adducted within 20 s. We therefore undertook to characterize the nature of LG adducts. Using liquid chromatography electrospray tandem mass spectrometry, we established that LGs form oxidized pyrrole adducts (lactams and hydroxylactams) with the epsilon amino group of lysine. Oxidation of low density lipoprotein resulted in readily detectable IsoLG adducts on apolipoprotein B after enzymatic digestion of the protein to individual amino acids. These studies identify a novel class of ketoaldehydes produced by the IsoP pathway that form covalent protein adducts at a rate that greatly exceeds that of other known aldehyde products of lipid peroxidation. Elucidation of the nature of the adducts formed by IsoLGs provides the basis to explore the formation of IsoLGs in vivo and investigate the potential biological ramifications of their formation in settings of oxidant injury. PMID- 10385609 TI - Adrenomedullary function is severely impaired in 21-hydroxylase-deficient mice. AB - Deficiency of 21-hydroxylase (21-OH), one of the most common genetic defects in humans, causes low glucocorticoid and mineralocorticoid production by the adrenal cortex, but the effect of this disorder on the adrenomedullary system is unknown. Therefore, we analyzed the development, structure, and function of the adrenal medulla in 21-OH-deficient mice, an animal model resembling human congenital adrenal hyperplasia. Chromaffin cells of 21-OH-deficient mice exhibited ultrastructural features of neuronal transdifferentiation with reduced granules, increased rough endoplasmic reticulum and small neurite outgrowth. Migration of chromaffin cells in the adrenal to form a central medulla was impaired. Expression of phenylethanolamine-N-methyltransferase (PNMT) was reduced to 27 +/- 9% (P<0.05), as determined by quantitative TaqMan polymerase chain reaction, and there was a significant reduction of cells staining positive for PNMT in the adrenal medulla of the 21-OH-deficient mice. Adrenal contents of epinephrine were decreased to 30 +/- 2% (P<0. 01) whereas norepinephrine and dopamine levels were reduced to 57 +/- 4% (P<0.01) and 50 +/- 9% (P<0.05), respectively. 21-OH deficient mice demonstrate severe adrenomedullary dysfunction, with alterations in chromaffin cell migration, development, structure, and catecholamine synthesis. This hitherto unrecognized mechanism may contribute to the frequent clinical, mental, and therapeutic problems encountered in humans with this genetic disease. PMID- 10385610 TI - Neogenesis vs. apoptosis As main components of pancreatic beta cell ass changes in glucose-infused normal and mildly diabetic adult rats. AB - We have investigated in adult rats made mildly diabetic by a low dose of streptozotocin (35 mg/kg; STZ rats) and in nondiabetic rats (ND rats) the mechanisms leading to adaptive changes in the beta cell mass, during glucose infusion and several days after stopping infusion. As early as 24 h of glucose infusion, the beta cell mass was maximally increased in ND and STZ rats. In both groups, this increase was due mainly to a rapid activation of neogenesis of new endocrine cells rather than to an increase in beta cell proliferation. Seven days after stopping glucose infusion, the beta cell mass returned to basal values in both groups as a result of stimulation of beta cell apoptosis and a decrease in beta cell replication rate. In glucose-infused ND rats, changes in the beta cell mass were correlated to insulin secretion, whereas in STZ rats, insulin secretion in response to glucose was still impaired whatever the beta cell mass. In conclusion, the data stress the impressive plasticity of the endocrine pancreas of adult rats. They also show that changes in beta cell mass in ND and STZ rats resulted from a disruption in the balance between neogenesis and apoptosis. PMID- 10385611 TI - Differential T cell response in central and peripheral nerve injury: connection with immune privilege. AB - The central nervous system (CNS), unlike the peripheral nervous system (PNS), is an immune-privileged site in which local immune responses are restricted. Whereas immune privilege in the intact CNS has been studied intensively, little is known about its effects after trauma. In this study, we examined the influence of CNS immune privilege on T cell response to central nerve injury. Immunocytochemistry revealed a significantly greater accumulation of endogenous T cells in the injured rat sciatic nerve than in the injured rat optic nerve (representing PNS and CNS white matter trauma, respectively). Use of the in situ terminal deoxytransferase-catalyzed DNA nick end labeling (TUNEL) procedure revealed extensive death of accumulating T cells in injured CNS nerves as well as in CNS nerves of rats with acute experimental autoimmune encephalomyelitis, but not in injured PNS nerves. Although Fas ligand (FasL) protein was expressed in white matter tissue of both systems, it was more pronounced in the CNS. Expression of major histocompatibility complex (MHC) class II antigens was found to be constitutive in the PNS, but in the CNS was induced only after injury. Our findings suggest that the T cell response to central nerve injury is restricted by the reduced expression of MHC class II antigens, the pronounced FasL expression, and the elimination of infiltrating lymphocytes through cell death. PMID- 10385612 TI - Functional heterogeneity of osteoclasts: matrix metalloproteinases participate in osteoclastic resorption of calvarial bone but not in resorption of long bone. AB - Data in the literature suggest that site-specific differences exist in the skeleton with respect to digestion of bone by osteoclasts. Therefore, we investigated whether bone resorption by calvarial osteoclasts (intramembranous bone) differs from resorption by long bone osteoclasts (endochondral bone). The involvement of two major classes of proteolytic enzymes, the cysteine proteinases (CPs) and matrix metalloproteinases (MMPs), was studied by analyzing the effects of selective low molecular weight inhibitors of these enzymes on bone resorption. Mouse tissue explants (calvariae and long bones) as well as rabbit osteoclasts, which had been isolated from both skeletal sites and subsequently seeded on bone slices, were cultured in the presence of inhibitors and resorption was analyzed. The activity of the CP cathepsins B and K and of MMPs was determined biochemically (CPs and MMPs) and enzyme histochemically (CPs) in explants and isolated osteoclasts. We show that osteoclastic resorption of calvarial bone depends on activity of both CPs and MMPs, whereas long bone resorption depends on CPs, but not on the activity of MMPs. Furthermore, significantly higher levels of cathepsin B and cathepsin K activities were expressed by long bone osteoclasts than by calvarial osteoclasts. Resorption of slices of bovine skull or cortical bone by osteoclasts isolated from long bones was not affected by MMP inhibitors, whereas resorption by calvarial osteoclasts was inhibited. Inhibition of CP activity affected the resorption by the two populations of osteoclasts in a similar way. We conclude that this is the first report to show that significant differences exist between osteoclasts of calvariae and long bones with respect to their bone resorbing activities. Resorption by calvarial osteoclasts depends on the activity of CPs and MMPs, whereas resorption by long bone osteoclasts depends primarily on the activity of CPs. We hypothesize that functionally different subpopulations of osteoclasts, such as those described here, originate from different sets of progenitors. PMID- 10385613 TI - Leptin induces oxidative stress in human endothelial cells. AB - Human umbilical vein endothelial cells (HUVEC) express functional receptors to leptin, the product of the ob gene. As human obesity is associated with atherosclerosis and hyperleptinemia, we investigated whether leptin, in addition to its angiogenic properties, exerts atherogenic effects through the generation of oxidative stress in endothelial cells. In HUVEC leptin increased the accumulation of reactive oxygen species (ROS), as assessed by the oxidation of 2', 7'- dichlorodihydrofluorescein, in a time- and concentration-dependent manner. In addition, leptin activated the NH2-terminal c-Jun kinase/stress activated protein kinase pathway as demonstrated by enhanced JNK activity and AP 1 DNA binding. Both effects were sensitive to antioxidant treatment with N acetylcysteine. NF-kappaB, another redox-sensitive transcription factor, was also activated by leptin stimulation in an oxidant-dependent manner. Finally, activation of both AP-1 and NF-kappaB was associated with an enhanced expression of the monocyte chemoattractant protein-1 in HUVEC. These findings demonstrate that ROS are second messengers involved in leptin-induced signaling in endothelial cells. Thus, chronic oxidative stress in endothelial cells under hyperleptinemia may activate atherogenic processes and contribute to the development of vascular pathology. PMID- 10385614 TI - Inhibition of caspase activity prevents CD95-mediated hepatic microvascular perfusion failure and restores Kupffer cell clearance capacity. AB - Using a murine model, we studied the effect of agonistic anti-CD95 antibodies (aCD95) on sinusoidal lining cells and a potential protection by caspase inhibition. C3H/HeN mice were intravenously administered aCD95 (10 microgram/mouse) or unspecific IgG (control) in the presence or absence of the caspase inhibitor z-VAD-fmk. Analysis of hepatic microcirculation using intravital fluorescence microscopy revealed severe (P<0.01) sinusoidal perfusion failure and reduced (P<0.05) phagocytic activity of Kupffer cells (KC) within 2 h. Transmission electron micrographs demonstrated loss of integrity of sinusoidal endothelial cells as early as 1 h after aCD95 application, whereas histological manifestation of hepatocellular apoptosis and hemorrhagic necrosis was most pronounced at 6 h. Blocking of caspase activity attenuated (P<0.01) both hepatic microvascular perfusion failure and KC dysfunction. Accordingly, full protection of the liver from apoptotic damage and intact microarchitecture was observed in histological sections after z-VAD-fmk treatment. Mortality rate was 40% 6 h after aCD95 administration, whereas all animals survived in the z-VAD-fmk group (P<0.05). The activation of caspases through CD95 may primarily lead to damage of sinusoidal endothelial cells and hepatic microvascular perfusion failure. Moreover, reduced phagocytic capacity of KC may contribute to accumulation of toxic metabolites released by dying cells at the local site of inflammation, further aggravating liver injury. PMID- 10385615 TI - A Sec7-related protein in Paramecium. AB - We have cloned and sequenced a SEC7-related gene in Paramecium tetraurelia that contains an open reading frame for 1135 amino acids encoding a 133 kDa protein, PSec7. Sec7, first identified in vesicular trafficking mutants in yeast, and its phylogenetic homologues function as guanine-nucleotide exchange factors for small G-proteins such as ARF (ADP-ribosylation factor). The deduced amino acid sequence in PSec7 for the motifs that form the ARF binding site are more than 70% identical to yeast Sec7 and similarly identical to ARNO, the human ARF exchange factor, with correct positioning of the critical glutamic acid residue within the motif region. Overall, the identity of PSec7 to yeast Sec7 is 32%. The deduced amino acid sequence also has five sequences that resemble IQ motifs, EF hand binding domains found in all myosins, and two pleckstrin homology domains. Similar sequences are present in yeast Sec7 and other Sec7-related molecules. A protein kinase A phosphorylation site may also be present. Southern blots suggest that a single gene encodes PSec7. Northern blots show that the message encoding PSec7 is induced on deciliation, followed by ciliogenesis, which suggests a role for PSec7 in cilia such as transport or targeting of ciliary membrane components. PMID- 10385616 TI - Central injection of nicotine increases hepatic and splenic interleukin 6 (IL-6) mRNA expression and plasma IL-6 levels in mice: involvement of the peripheral sympathetic nervous system. AB - Accumulating evidence suggests that plasma levels of interleukin 6 (IL-6), a major cytokine stimulating the synthesis of acute-phase proteins, are intimately regulated by the central nervous system. Nicotine, one of the major drugs abused by humans, has been shown to affect immunological functions. In the present study, effects of intracerebroventricular (i.c.v.) injection of nicotine on plasma IL-6 levels were investigated in mice. Nicotine administered i.c.v. dose dependently increased plasma IL-6 levels; the lowest effective dose was 0.3 ng/mouse and the maximal effect was attained with the dose of 105 ng/mouse. The nicotine (105 ng/mouse, i.c.v.)-induced plasma IL-6 levels peaked at 3 h and approached basal levels 6 h after injection. Mecamylamine, a nicotinic receptor antagonist, blocked nicotine-induced plasma IL-6 levels. Depletion of peripheral norepinephrine with 6-hydroxydopamine [100 mg/kg, intraperitoneal (i. p.)] inhibited the nicotine-induced plasma IL-6 levels by 57%, whereas central norepinephrine depletion with 6-hydroxydopamine (50 microgram/mouse, i.c.v.) had no effect. Pretreatment with prazosin (alpha1-adrenergic antagonist; 1 mg/kg, i.p.), yohimbine (alpha2-adrenergic antagonist; 1 mg/kg, i.p.), and ICI-118,551 (beta2-adrenergic antagonist; 2 mg/kg, i.p.), but not with betaxolol (beta1 adrenergic antagonist; 2 mg/kg, i.p.), inhibited nicotine-induced plasma IL-6 levels. Among the peripheral organs, including the pituitary, adrenals, heart, lung, liver, spleen, and lymph nodes, nicotine (105 ng/mouse, i.c.v.) increased IL-6 mRNA expression only in the liver and spleen, which was inhibited by peripheral norepinephrine depletion. These results suggest that stimulation of central nicotinic receptors induces plasma IL-6 levels and IL-6 mRNA expression in the liver and spleen via the peripheral sympathetic nervous system, alpha1-, alpha2-, and beta2-adrenoreceptors being involved. PMID- 10385617 TI - Porcine relaxin, a 500 million-year-old hormone? the tunicate Ciona intestinalis has porcine relaxin. AB - The fossil record of tunicates reaches back to the upper Cambrian period. Ascidians have mobile, tadpole-like juvenile forms with a notochord, which inspired the classification of tunicates as Urochordata, i.e., predecessors of vertebrates. The genome of the tunicate Ciona intestinalis contains a relaxin coding region that is organized like a mammalian gene, i.e., signal peptide, B chain domain, connecting peptide domain, followed by the A-chain domain with a stop codon after cysteine A-22. RNA-derived cDNA encodes a relaxin that is identical to the circulating form of the porcine hormone. In contrast to the porcine gene, the ascidian gene has no intron in the C-peptide domain, and in that respect is similar to the bombyxin gene of the silkworm. During the spawning period, only enough relaxin could be extracted and isolated from gonads of C. intestinalis for a partial sequence analysis. Remarkable as it may be, these findings suggest that relaxin is identical in pigs, whales, and the tunicate C. intestinalis. PMID- 10385618 TI - CDK inhibitors: positive and negative regulators of G1-phase progression. PMID- 10385619 TI - Developmental regulation of SR protein phosphorylation and activity. AB - Serine/arginine-rich splicing factors (SR proteins) are substrates for serine phosphorylation that can regulate SR protein function. We have observed gross changes in SR protein phosphorylation during early development coincident with major zygotic gene activation in the nematode Ascaris lumbricoides. These differences correlate with large-scale changes in SR protein activity in promoting both trans- and cis-splicing. Importantly, inactive early stage extracts can be made splicing competent on addition of later stage SR proteins. These data suggest that changes in SR protein phosphorylation have a role in the activation of pre-mRNA splicing during early development. PMID- 10385620 TI - Brachyury downstream notochord differentiation in the ascidian embryo. AB - The ascidian tadpole represents the most simplified chordate body plan. It contains a notochord composed of just 40 cells, but as in vertebrates Brachyury is essential for notochord differentiation. Here, we show that the misexpression of the Brachyury gene (Ci-Bra) of Ciona intestinalis is sufficient to transform endoderm into notochord. Subtractive hybridization screens were conducted to identify potential Brachyury target genes that are induced upon Ci-Bra misexpression. Of 501 independent cDNA clones that were surveyed, 38 were specifically expressed in notochord cells. These potential Ci-Bra downstream genes appear to encode a broad spectrum of divergent proteins associated with notochord formation. PMID- 10385621 TI - Targeting genes for self-excision in the germ line. AB - A procedure is described that directs the self-induced deletion of DNA sequences as they pass through the male germ line of mice. The testes-specific promoter from the angiotensin-converting enzyme gene was used to drive expression of the Cre-recombinase gene. Cre was linked to the selectable marker Neor, and the two genes flanked with loxP elements. This cassette was targeted to the Hoxa3 gene in mouse ES cells that were in turn used to generate chimeric mice. In these chimeras, somatic cells derived from the ES cells retained the cassette, but self excision occurred in all ES-cell-derived sperm. PMID- 10385622 TI - ACF consists of two subunits, Acf1 and ISWI, that function cooperatively in the ATP-dependent catalysis of chromatin assembly. AB - The assembly of core histones and DNA into periodic nucleosome arrays is mediated by ACF, an ISWI-containing factor, and NAP-1, a core histone chaperone, in an ATP dependent process. We describe the isolation of Drosophila acf1 cDNA, which encodes the p170 and p185 forms of the Acf1 protein in ACF. Acf1 is a novel protein that contains two PHD fingers, one bromodomain, and two new conserved regions. Human WSTF, which is encoded by one of multiple genes that is deleted in Williams syndrome individuals, is the only currently known mammalian protein with each of the conserved motifs in Acf1. Purification of the native form of Acf1 led to the isolation of ACF comprising Acf1 (both p170 and p185 forms) and ISWI. Native Acf1 did not copurify with components of NURF or CHRAC, which are other ISWI-containing complexes in Drosophila. Purified recombinant ACF, consisting of Acf1 (either p185 alone or both p170 and p185) and ISWI, catalyzes the deposition of histones into extended periodic nucleosome arrays. Notably, the Acf1 and ISWI subunits function synergistically in the assembly of chromatin. ISWI alone exhibits a weak activity that is approximately 3% that of ACF. These results indicate that both Acf1 and ISWI participate in the chromatin assembly process and suggest further that the Acf1 subunit confers additional functionality to the general 'motor' activity of ISWI. PMID- 10385623 TI - A protein phosphatase functions to recycle RNA polymerase II. AB - Transcription is regulated by the state of phosphorylation of a heptapeptide repeat known as the carboxy-terminal domain (CTD) present in the largest subunit of RNA polymerase II (RNAPII). RNAPII that associates with transcription initiation complexes contains an unphosphorylated CTD, whereas the elongating polymerase has a phosphorylated CTD. Transcription factor IIH has a kinase activity specific for the CTD that is stimulated by the formation of a transcription initiation complex. Here, we report the isolation of a cDNA clone encoding a 150-kD polypeptide, which, together with RNAPII, reconstitutes a highly specific CTD phosphatase activity. Functional analysis demonstrates that the CTD phosphatase allows recycling of RNAPII. The phosphatase dephosphorylates the CTD allowing efficient incorporation of RNAPII into transcription initiation complexes, which results in increased transcription. The CTD phosphatase was found to be active in ternary elongation complexes. Moreover, the phosphatase stimulates elongation by RNAPII; however, this function is independent of its catalytic activity. PMID- 10385624 TI - Activation of topoisomerase II-mediated excision of chromosomal DNA loops during oxidative stress. AB - Hydrogen peroxide (H2O2), a reactive oxygen species (ROS), is known to induce oxidative stress and apoptosis. U937 cells treated with H2O2 were shown to produce high molecular weight (HMW) DNA fragments approximately 50 to 100 kb in size in <1 min. The formation of these HMW DNA fragments is reversible and shown to be mediated by DNA topoisomerase II (TOP2). Following this initial event, formation of irreversible HMW DNA fragments and nucleosomal ladders occurs. Our results thus demonstrate a potential role of TOP2 in oxidative damage of DNA and apoptotic cell death. PMID- 10385626 TI - Asymmetric and node-specific nodal expression patterns are controlled by two distinct cis-acting regulatory elements. AB - The TGFbeta-related molecule Nodal is required for establishment of the anterior posterior (A-P) and left-right (L-R) body axes of the vertebrate embryo. In mouse, several discrete sites of nodal activity closely correlate with its highly dynamic expression domains. nodal function in the posterior epiblast promotes primitive streak formation, whereas transient nodal expression in the extraembryonic visceral endoderm is essential for patterning the rostral central nervous system. Asymmetric nodal expression in the developing node and at later stages in left lateral plate mesoderm has been implicated as a key regulator of L R axis determination. We have analyzed the cis-regulatory elements controlling nodal expression domains during early development. We show that the regulatory sequences conferring node-specific expression are contained in an upstream region of the locus, whereas early expression in the endoderm and epiblast and asymmetric expression at later stages on the left side of the body axis are controlled by a 600-bp intronic enhancer. Targeted deletion of a 100-bp subregion of this intronic enhancer eliminates nodal expression in the early epiblast and visceral endoderm and disrupts asymmetric expression in the node and lateral plate mesoderm. Thus, developmentally regulated nodal expression at distinct tissue sites during A-P and L-R axis formation is potentially controlled by common transcriptional activators. PMID- 10385625 TI - Cyclo-oxygenase-2-derived prostacyclin mediates embryo implantation in the mouse via PPARdelta. AB - We have demonstrated previously that cyclo-oxygenase-2 (COX2), the rate-limiting enzyme in the biosynthesis of prostaglandins (PGs), is essential for blastocyst implantation and decidualization. However, the candidate PG(s) that participates in these processes and the mechanism of its action remain undefined. Using COX2 deficient mice and multiple approaches, we demonstrate herein that COX2-derived prostacyclin (PGI2) is the primary PG that is essential for implantation and decidualization. Several lines of evidence suggest that the effects of PGI2 are mediated by its activation of the nuclear hormone receptor PPARdelta, demonstrating the first reported biologic function of this receptor signaling pathway. PMID- 10385627 TI - Determination of left/right asymmetric expression of nodal by a left side specific enhancer with sequence similarity to a lefty-2 enhancer. AB - The nodal gene is expressed on the left side of developing mouse embryos and is implicated in left/right (L-R) axis formation. The transcriptional regulatory regions of nodal have now been investigated by transgenic analysis. A node specific enhancer was detected in the upstream region (-9.5 to -8.7 kb) of the gene. Intron 1 was also shown to contain a left side-specific enhancer (ASE) that was able to direct transgene expression in the lateral plate mesoderm and prospective floor plate on the left side. A 3. 5-kb region of nodal that contained ASE responded to mutations in iv, inv, and lefty-1, all genes that act upstream of nodal. The same 3. 5- kb region also directed expression in the epiblast and visceral endoderm at earlier stages of development. Characterization of deletion constructs delineated ASE to a 340-bp region that was both essential and sufficient for asymmetric expression of nodal. Several sequence motifs were found to be conserved between the nodal ASE and the lefty-2 ASE, some of which appeared to be essential for nodal ASE activity. These results suggest that similar transcriptional mechanisms underlie the asymmetric expression of nodal and of lefty-2 as well as the earlier expression of nodal in the epiblast and endoderm. PMID- 10385628 TI - Interaction between FGF and BMP signaling pathways regulates development of metanephric mesenchyme. AB - Nephrogenesis in the mouse kidney begins at embryonic day 11 and ends approximately 10 days postpartum. During this period, new nephrons are continually being generated from a stem-cell population-the nephrogenic mesenchyme-in response to signals emanating from the tips of the branching ureter. Relatively little is known about the mechanism by which the nephrogenic mesenchyme cell population is maintained at the tips of the ureter in the presence of signals promoting tubulogenesis. Previous studies have shown that a loss of Bmp7 function leads to kidney defects that are a likely result of progressive loss of nephrogenic mesenchyme by apoptosis. The studies presented here demonstrate that BMP7 signaling can prevent apoptosis in explants of metanephric mesenchyme. The surviving mesenchyme cell population, however, is not competent to respond to signals promoting tubulogenesis. In conjunction with FGF2, BMP7 promotes growth and maintains competence of the mesenchyme in vitro. In addition, FGF2 and BMP7 signaling, both independently and in combination, inhibit tubulogenesis. Interestingly, FGF2 and BMP7 also promote expansion of the stromal progenitor cell population in whole kidney culture. Because BMP7 is not produced by stromal progenitor cells, these data suggest a novel interaction between the nephrogenic mesenchyme and stromal progenitor cell populations. A model for the regulation of nephrogenesis by FGF and BMP signaling is presented. PMID- 10385629 TI - Cdc53/cullin and the essential Hrt1 RING-H2 subunit of SCF define a ubiquitin ligase module that activates the E2 enzyme Cdc34. AB - SCFCdc4 (Skp1, Cdc53/cullin, F-box protein) defines a family of modular ubiquitin ligases (E3s) that regulate diverse processes including cell cycle, immune response, and development. Mass spectrometric analysis of proteins copurifying with Cdc53 identified the RING-H2 finger protein Hrt1 as a subunit of SCF. Hrt1 shows striking similarity to the Apc11 subunit of anaphase-promoting complex. Conditional inactivation of hrt1(ts) results in stabilization of the SCFCdc4 substrates Sic1 and Cln2 and cell cycle arrest at G1/S. Hrt1 assembles into recombinant SCF complexes and individually binds Cdc4, Cdc53 and Cdc34, but not Skp1. Hrt1 stimulates the E3 activity of recombinant SCF potently and enables the reconstitution of Cln2 ubiquitination by recombinant SCFGrr1. Surprisingly, SCF and the Cdc53/Hrt1 subcomplex activate autoubiquitination of Cdc34 E2 enzyme by a mechanism that does not appear to require a reactive thiol. The highly conserved human HRT1 complements the lethality of hrt1Delta, and human HRT2 binds CUL-1. We conclude that Cdc53/Hrt1 comprise a highly conserved module that serves as the functional core of a broad variety of heteromeric ubiquitin ligases. PMID- 10385631 TI - Nitric oxide in liver injury. PMID- 10385630 TI - Somatic pairing of homologs in budding yeast: existence and modulation. AB - FISH analysis of well-spread chromosomes reveals that homologs are paired in vegetatively growing budding yeast diploid cells, via multiple interstitial interactions, and independent of recA homologs and mating type heterozygosity. Pairing is present during G1 and G2, and in cells arrested at G1 by mating pheromone, but is disrupted during S phase. Thus, somatic pairing is qualitatively analogous to premeiotic and early meiotic pairing. S-phase pairing disruption occurs by a complex intranuclear program involving regional, nucleus wide, and temporal determinants. Pairing is also disrupted in two G2-arrest conditions (cdc13ts and nocodazole). Together these findings suggest that cell cycle signals may provoke pairing disruption by modulating underlying chromosome and/or chromatin structure. Whether the cell chooses to disrupt pairing contacts or not (e.g., S phase and G2 arrest, but not G1 arrest or normal G1 or G2), could be dictated by functional considerations involving homolog/sister discrimination. PMID- 10385632 TI - Long-term ursodeoxycholic acid therapy is associated with reduced risk of biliary pain and acute cholecystitis in patients with gallbladder stones: a cohort analysis. AB - Whether ursodeoxycholic acid (UDCA) therapy alters the long-term clinical course of gallstones (GS) without stone dissolution remains unknown. We aimed to clarify the relationship between long-term UDCA therapy and risks of biliary pain or acute cholecystitis in GS patients. We also aimed to identify factors affecting the natural course, and to explore a simple patient selection criteria for UDCA therapy. A cohort of 527 uncomplicated GS patients with or without UDCA (600 mg/d) followed for up to 18 years was analyzed. Patients who had frequent attacks or were complicated with cholecystitis were converted to cholecystectomy. History and UDCA therapy were identified on Cox analysis as 2 factors affecting the long term clinical course. In patients without therapy, history was the only predictor of biliary pain among various patient or stone characteristics; biliary pain was rare in asymptomatic patients, while frequent in symptomatic patients (P <.001). UDCA therapy was associated with reduced risk for biliary pain in both symptomatic (62% vs. 92% in untreated patients at 10 years; P <.001; relative risk, 0.19; 95% CI, 0.10-0.34) and asymptomatic patients (6% vs. 12% in untreated patients at 10 years; P =.037; relative risk, 0.19; 95% CI, 0.04-0.91). Risk for the conversion was also reduced in UDCA-treated symptomatic patients (26% vs. 88% in untreated patients at 10 years, P <.001; relative risk, 0.08; 95% CI, 0.03 0.22). These effects were independent of stone dissolution. Three factors were identified on Cox analysis as affecting GS dissolution: radiolucency, small size (<10 mm) of stones, and visualized gallbladder (GB) on cholecystogram. A selection criteria based on these appears to exhibit high sensitivity (74%) and specificity (95%) for dissolution. UDCA therapy might be considered in symptomatic patients fulfilling these criteria, and also in patients who have significant surgical risk, because the longterm therapy is clearly associated with reduced risk of biliary pain and acute cholecystitis. PMID- 10385633 TI - The pravastatin-induced decrease of biliary cholesterol secretion is not directly related to an inhibition of cholesterol synthesis in humans. AB - 3-Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors have been reported to suppress biliary cholesterol secretion and saturation. It remains unproven whether this is mediated by inhibition of cholesterol synthesis. Therefore, the effect of a long-term administration of pravastatin on cholesterogenesis and on biliary lipid secretion was investigated in seven healthy volunteers. Placebo or 40 mg of pravastatin were taken daily at bedtime for 5 weeks using a double-blind crossover design. During the last week, 12 hours after the last drug intake 0.04 mmol [1-13C]acetate/kg. h and 0.5 g polyethylene glycol 4,000/h were infused intraduodenally for 15 hours. Plasma and duodenal bile samples were collected hourly. Thereafter, the decay of [13C]labeled plasma cholesterol was measured during the following 3 days. The fractional and absolute syntheses of plasma and biliary cholesterol were determined by gas chromatography mass spectrometry using mass isotopomer distribution analysis. At the end of the pravastatin period plasma total and low-density lipoprotein (LDL) cholesterol had decreased by 20% and 24%, respectively. Similarly, pravastatin suppressed biliary secretion rates of cholesterol, total bile acids and phospholipids (P <.05) by 46%, 36%, and 51%. As a consequence, cholesterol saturation index remained unchanged. However, fractional syntheses of cholesterol were comparable (P >.05) on placebo compared with pravastatin with 3.1% versus 4.0% in plasma and 4.3% versus 5.2% in bile after 15 hours, respectively. The mean absolute synthesis rates amounted to 0.3 mg/kg/h on placebo versus 0.4 on pravastatin (P >. 05). In conclusion, the pravastatin-induced reduction of biliary cholesterol secretion is not directly related to an inhibition of cholesterol synthesis. PMID- 10385634 TI - Characterization of the autoantibody responses to recombinant E3 binding protein (protein X) of pyruvate dehydrogenase in primary biliary cirrhosis. AB - Autoantibodies to the pyruvate dehydrogenase complex (PDC) are present in the serum of more than 95% of patients with primary biliary cirrhosis (PBC), the major epitope being the inner lipoyl domain of the E2 component. Immunoblotting suggests a similar prevalence of antibodies to a tightly associated lipoic acid containing protein, E3 binding protein (E3BP). Attempts to resolve E3BP from E2 have been unsuccessful, restricting study of the nature and significance of antibody responses to the individual proteins. In particular, it is unclear (1) whether there is true cross-reactivity between E3BP and E2 and, if so, which is the originating response and (2) whether autoantibodies preferentially bind a lipoylated epitope on E3BP as is the case with PDC-E2. In this study, complementary DNAs encoding rE2, full-length rE3BP, its single lipoyl domain (rLip), and core domain (rE3BPCore) were cloned, and the proteins were expressed in Escherichia coli. Sera from 47 PBC patients were studied by immunoblotting and enzyme-linked immunosorbent assay (ELISA) against rE2, rE3BP, rE3BPCore, and both unlipoylated (U) and lipoylated (L) rLip. All sera were reactive by ELISA to some degree with all recombinant proteins except rE3BPCore, to which only 6 of 47 showed any reactivity. Significant correlations (P <.0001) were observed when comparing absorbance values for rE3BP with both rLip (U) (r = 0.793) and (L) (r = 0.963). The mean absorbance for rLip (U, 0.26 +/- 0.05) was, however, significantly lower than the absorbance for rLip (L) (0.78 +/- 0.12; P <.0001). After probing by immunoblotting and elution of antibodies from rE2 and rE3BP, subsequent reprobing against the components in whole PDC revealed true cross reactivity. In summary, the response to E3BP is primarily directed against the lipoylated domain of the protein. It still remains unclear, however, whether the initial breakdown of tolerance is to E2 or E3BP. PMID- 10385635 TI - Liver and bile duct pathology following Cryptosporidium parvum infection of immunodeficient mice. AB - Patients with acquired immune deficiency syndrome (AIDS) and boys with mutations of the CD154 gene (causing congenital X-linked immunodeficiency with hyper-IgM [XHIM]) are susceptible to chronic infections of the biliary tract with Cryptosporidium parvum (CP) that may lead to biliary sclerosis and ultimately to cholangiocarcinoma. To determine whether the CP infection and the consequent immune response contribute independently to this morbidity, we infected mice with severe combined immunodeficiency (SCID) or with disrupted genes for CD154, CD40, or interferon gamma (IFN-gamma) with CP. Even when CP infection persisted for 16 weeks, the SCID mice developed only mild triaditis, without apoptosis of biliary epithelial cells (BEC). Fifty percent of the CD154 knockout mice developed lobular hepatitis with acute and chronic triaditis. The CD40 knockout mice developed marked triaditis, and the IFN-gamma knockouts either succumbed to enteritis or survived to develop marked triaditis, portal fibrosis, biliary sclerosis, necrosis with dilation of duct-like structures within the porta hepatis, and dysplastic changes. CP-infected SCID mice reconstituted with T cells from IFN-gamma knockout donors either developed severe enteritis or survived to develop triaditis, cholangitis, lobular hepatitis with periductular sclerosis, and scarring. Mice with disruptions of both the CD40 and IFN-gamma genes remained infected by CP and developed bile duct and liver disease, but not enteritis. Our results suggest that T-cell cytokines are required for the inflammatory and sclerosing responses to CP infection in immunodeficient animals. The response of immunodeficient mice to CP infection may model at least the initial steps toward the development of sclerosing cholangitis or bile duct cancers in XHIM patients. PMID- 10385636 TI - In situ nucleic acid detection of PDC-E2, BCOADC-E2, OGDC-E2, PDC-E1alpha, BCOADC E1alpha, OGDC-E1, and the E3 binding protein (protein X) in primary biliary cirrhosis. AB - The characteristic serological feature of primary biliary cirrhosis (PBC) is the presence of antimitochondrial antibodies (AMAs), and the major proteins recognized by AMAs are subunits of the 2-oxo acid dehydrogenase complexes (2 OADC), including the E2 components of the pyruvate dehydrogenase complex (PDC), the 2-oxo-glutarate dehydrogenase complex (OGDC), the branched-chain 2-oxoacid dehydrogenase complex (BCOADC), the E3 binding protein (E3BP or protein X) and the E1a component of mammalian PDC. Previous work has postulated that either E3BP, or a molecule cross-reactive with the PDC-E2 molecule, is uniquely expressed on the surface of biliary epithelial cells in PBC. To address this issue, we performed in situ hybridization for all of the major 2-OADC components at the mRNA level, including PDC-E2, BCOADC-E2, OGDC-E2, PDC-E1a, BCOADC-E1a, OGDC-E1, and E3BP using 13 PBC and 9 control livers using 7 mitochondrial antisense probes. In both PBC and controls, the expression of all 2-OADC component mRNA studied herein were found in hepatocytes and infiltrating mononuclear cells, without significant differences. Interestingly, however, despite published data on immunohistochemical staining, interlobular bile ducts including the injured bile ducts in PBC were generally negative or only faintly positive, with the exception of 1 bile duct in 1 of 13 cases of PBC and 1 of 9 control liver specimens. Moreover, confocal microscopic examination and image analysis revealed that the mRNA signal intensity of each of the 2-OADC components in the bile ducts of PBC was relatively lower in comparison with control liver diseases. These data suggest that continuous enhanced synthesis of the 2-OADC components is not likely to be occurring in the biliary epithelial cells in PBC, and that an increase of PDC-E2 or E3BP immunoreactivity in PBC is caused by exogenous imported or cross-reactive molecules. PMID- 10385637 TI - The effect of mechanically enhancing portal venous inflow on hepatic oxygenation, microcirculation, and function in a rabbit model with extensive hepatic fibrosis. AB - Enhancing the portal venous blood flow (PVBF) has been shown to reduce portal pressure and intrahepatic vascular resistance and to improve liver function in isolated cirrhotic rodent livers in vitro. The aim of this study was to assess the short-term effect of mechanically pumping the portal inflow on hepatic microcirculation (HM), oxygenation, and function in an animal model of extensive hepatic fibrosis. New Zealand white rabbits underwent laparotomy and exposure of the liver: group 1 (n = 7) were normal controls; group 2 (n = 7) had hepatic fibrosis. Total hepatic blood flow (THBF) and HM was measured along with continuous monitoring of intrahepatic tissue oxygenation using near infrared spectroscopy (NIRS). Baseline hepatic hemodynamics and liver function were measured in both groups. PVBF was then increased by 50% over a 3-hour period in the hepatic fibrosis group using a miniature portal pump designed for human implantation, and the hemodynamics were monitored continuously. Liver function tests were repeated after portal pumping. In comparison with normal controls, animals with hepatic fibrosis had a higher portal pressure (13.0 +/- 3.6 vs. 3.7 +/- 1.4 mm Hg, P <.001, mean +/- SD vs. controls), reduced PVBF (52.4 +/- 24.6 vs. 96.9 +/- 21.1 mL/min, P =.003), and increased portal vascular resistance (P =. 001). THBF and flow in the HM was lower than in controls, and liver function tests were abnormal. After a 3-hour period of enhanced portal flow in animals with hepatic fibrosis, the portal pressure greatly reduced (13.0 +/- 3.6 to 2.5 +/- 1.1 mm Hg, P <.001) as did the intrahepatic portal resistance (0.32 +/- 0.18 to 0.04 +/- 0.03 mm Hg/mL/min, P =.006). Flow in the HM improved (143 +/- 16 to 173 +/- 14 flux units, P =.006) and was associated with improved hepatic tissue oxygenation, tissue oxy-hemoglobin (HbO2) and cytochrome oxidase being increased by 24.4 +/- 7.5 and 5.65 +/- 2.30 micromol/L above the baseline value (P <.001), respectively. A 3-hour period of mechanical portal pumping produced a dramatic improvement in liver function, bilirubin (41.1 +/- 25.9 to 10.0 +/- 5.9 micromol/L, P =. 040), aspartate transaminase (AST) (135.5 +/- 52.3 to 56.3 +/- 19.8 U/L, P =.006) and lactate dehydrogenase (LDH) (2,030.1 +/- 796.3 to 1,309.8 +/- 431.6 IU/L, P =.006; prepumping vs. postpumping, all P <. 050). In conclusion, portal pumping in this rabbit model with extensive hepatic fibrosis improved liver parenchymal perfusion, oxygenation, and function. PMID- 10385638 TI - Development of autoimmune hepatitis following liver transplantation for primary biliary cirrhosis. AB - Two patients undergoing liver transplantation for classical end-stage primary biliary cirrhosis (PBC) are described, who went on to develop de novo autoimmune hepatitis (AIH) in the transplanted liver. The presentation, in both instances, was with malaise and lethargy. Markedly elevated serum transaminases were found, together with a raised serum IgG and/or globulin fraction and histological features on liver biopsy typical of AIH. Both cases had had changes in their immunosuppressive therapy before the onset of AIH episodes, and both rapidly responded to reinstitution of steroid therapy. The finding, in each case, of a coincidental multiple HLA class I allele match between the recipient and their liver donor suggests that HLA class I-restricted mechanisms may play an important role in the pathogenesis of AIH. PMID- 10385639 TI - Systemic and splanchnic hemodynamic changes after liver transplantation for cirrhosis: a long-term prospective study. AB - The effect of orthotopic liver transplantation (OLT) on the systemic and splanchnic hemodynamic alterations of cirrhosis is still largely unknown. The aim of this study was to prospectively investigate the long-term changes induced by OLT on several hemodynamic parameters. In 28 patients undergoing OLT for cirrhosis, the following parameters were measured before surgery and subsequently at 6-month intervals (mean follow-up period, 17 months): cardiac index, mean arterial pressure (MAP), heart rate, total peripheral resistance (TPR), portal vein flow velocity and flow volume, spleen size, and Doppler ultrasound resistance or pulsatility indexes (RI or PI) in the: 1) interlobular renal, 2) superior mesenteric, 3) splenic, and 4) hepatic arteries. The same parameters were measured in 10 healthy controls. After OLT, cardiac index and heart rate significantly decreased (P <.01), while MAP and TPR increased (P <.001), so that any significant difference from controls disappeared. Renal RI progressively decreased, achieving a significant reduction (P <.05) to normal values at the 12th month of follow-up. Portal flow velocity and hepatic and splenic RI returned to values not significantly different from controls. Portal flow volume increased over normal values after OLT (P <.001), and SMA PI, lower than normal before OLT, did not show any statistically significant increase thereafter. Spleen size decreased significantly, but persisted to be larger than in controls. In conclusion, systemic, renal, and most, but interestingly not all, splanchnic circulatory alterations of cirrhosis are restored to normal after OLT. PMID- 10385640 TI - Breakdown of tolerance to pyruvate dehydrogenase complex in experimental autoimmune cholangitis: a mouse model of primary biliary cirrhosis. AB - The autoimmune liver disease primary biliary cirrhosis (PBC) is characterized by autoreactive responses to a highly conserved self-antigen, pyruvate dehydrogenase complex (PDC). We recently reported the development of PBC-like lesions in SJL mice sensitized with PDC and have named this model disease experimental autoimmune cholangitis (EAC). In the present study, the breakdown of tolerance to PDC has been investigated in animals sensitized for EAC. Splenic mononuclear cells from SJL mice sensitized with bovine heart PDC (bPDC) in adjuvant showed T cell proliferative and mixed Th1/Th2 cytokine secretory responses following in vitro stimulation with bPDC. Despite the likelihood of extensive sequence homology with mouse PDC (there is a greater than 95% sequence identity between rat and human PDC-E2 subunits), bPDC was highly immunogenic inducing significant T- and B-cell responses in the absence of any form of adjuvant. The multi-subunit quaternary structure of intact PDC was critical for this immunostimulatory activity because no response was produced by sensitization with monomeric recombinant PDC-E2 inner lipoyl domain. Mice sensitized with bPDC and CFA developed, within 2 weeks of sensitization, high-titer antibody responses reactive with bPDC that were fully cross-reactive with the murine homologue. Breakdown of T-cell tolerance to self-PDC took significantly longer, not being seen until 20 weeks postsensitization; a similar length of time to that previously shown to be required for EAC lesion development. Conclusions drawn from these data may have important implications for our understanding, and therapeutic manipulation, of PBC in humans. PMID- 10385641 TI - Ursodeoxycholic acid inhibits eosinophil degranulation in patients with primary biliary cirrhosis. AB - Eosinophilia is a distinctive feature of primary biliary cirrhosis (PBC), especially in its early stages. Intriguingly, treatment with ursodeoxycholic acid (UDCA) ameliorates eosinophilia as well as liver tests in patients with PBC. It remains unknown, however, whether eosinophils in PBC patients are functionally activated and whether UDCA inhibits eosinophil activation. In the present study, we systematically examined eosinophil dynamics in the blood and liver in patients with stage I to II PBC before and after UDCA treatment. We determined serum concentrations of eosinophil granule proteins (major basic protein [MBP] and eosinophil-derived neurotoxin [EDN]) by radioimmunoassay and quantitated eosinophil degranulation using computer-assisted morphometry after MBP immunohistochemistry. Before UDCA treatment, patients with PBC (n = 25) showed significantly higher circulating eosinophil counts (P <. 05) and serum concentrations of MBP (P <.0005) and EDN (P <.02) compared with patients with chronic viral hepatitis (n = 22), autoimmune hepatitis (n = 10), and obstructive jaundice (n = 12). Four-week UDCA treatment significantly reduced blood eosinophil counts (P <.0001) and serum MBP (P <.0001) and EDN (P <.0001) levels in PBC patients. MBP immunohistochemistry and computer-assisted quantitative morphometry showed infiltration and degranulation of eosinophils in the portal tract in patients with PBC and significant reductions in the number of sites and the area occupied by extracellular MBP deposits after UDCA treatment for 2 years (P <.02) but not in placebo-treated patients. Our results suggest that eosinophils in patients with PBC are not only increased in number, but also release granule proteins, and that UDCA treatment inhibits this eosinophil activation/degranulation. PMID- 10385642 TI - Carvedilol, a new nonselective beta-blocker with intrinsic anti- Alpha1 adrenergic activity, has a greater portal hypotensive effect than propranolol in patients with cirrhosis. AB - Only some patients show a substantial hepatic venous pressure gradient (HVPG) reduction after propranolol, which makes it desirable to investigate drugs with greater portal hypotensive effect. The aim of this study was to investigate whether carvedilol, a nonselective beta-blocker with anti-alpha1-adrenergic activity, may cause a greater HVPG reduction than propranolol. Thirty-five cirrhotic patients had hemodynamic measurements before and after the random administration of carvedilol (n = 14), propranolol (n = 14), or placebo (n = 7). Carvedilol markedly reduced HVPG, from 19.5 +/- 1.3 to 15.4 +/- 1 mm Hg (P <.0001). This HVPG reduction was greater than after propranolol (-20.4 +/- 2 vs. 12.7 +/- 2%, P <.05). Moreover, carvedilol decreased HVPG greater than 20% of baseline values or to . 1). TTV group 2 infection was identified in 16 cases (61.5%) and 4 controls (36.4%) (P >.1) using a type-specific PCR method. Sequence analysis of 222 nt of TTV DNA demonstrated that the remaining 10 cases and 7 controls were all infected by group 1. The odds ratio (OR) for TTV-DNA positivity, adjusted for demographic variables, hepatitis B surface antigen (HBsAg), hepatitis C virus (HCV) RNA, and heavy alcohol intake was 1.8 (95% CI: 0.7-4.8; P >.1). The OR did not change when the analysis was restricted to 14 HCC cases and 56 controls who were negative for each known risk factor for HCC (OR = 1.7; 95% CI: 0.8-4.0). TTV-DNA positivity was not associated with transfusion history. The prevalence of TTV DNA was higher among HCC cases positive for HBsAg (10 of 38 [26.3%]) than among those positive for HCV RNA (8 of 62 [12.9%]) or negative for hepatitis B virus (HBV), HCV, and hepatitis G virus (HGV) infections (5 of 62 [8. 1%]) (P =.02). This study does not support the hypothesis of an association between TTV infection and HCC. PMID- 10385671 TI - Intracellular single-chain antibody against hepatitis B virus core protein inhibits the replication of hepatitis B virus in cultured cells. AB - Hepatitis B virus (HBV) is one of the major causes of chronic liver diseases and hepatocellular carcinoma. In this study, we used a single chain antibody (sFv), which is a man-made antibody with a strong affinity of immunoglobulin, to inhibit HBV replication. Because HBV replication can only take place in the viral nucleocapsid made of HBV core protein (HBc), we generated anti-HBc sFv and examined whether intracellular anti-HBc sFv could inhibit viral replication in the human hepatoblastoma-derived cell line that produces HBV (HB611). With respect to HBV replication intermediates, both single-stranded and partially double-stranded DNA intermediates were markedly suppressed in the cells expressing anti-HBc sFv, although HBV RNA intermediates were not affected. This suggested that intracellular anti-HBc sFv inhibited HBV DNA replication by inhibiting reverse transcription from HBV pregenome RNA to single-stranded DNA. Because the sFv-HBc complex was detected in the cells expressing anti-HBc sFv by immunoprecipitation analysis but the quantity of intracellular HBc was not affected, the anti-HBc sFv was suggested to inhibit HBV DNA replication by interfering with the function of HBc. These results indicate that intracellular sFv against HBc might be effective as a novel active molecule for gene therapy of hepatitis B. PMID- 10385672 TI - Dominant negative mutants of the duck hepatitis B virus core protein interfere with RNA pregenome packaging and viral DNA synthesis. AB - Dominant negative (DN) mutants of the hepadnaviral core protein are potent inhibitors of viral replication. We have previously shown that fusion of sequences derived from the duck hepatitis B virus (DHBV) polymerase (Pol), DHBV small surface protein (S), bacterial beta-galactosidase (lacZ), or green fluorescent protein (GFP) to the carboxy terminus of the DHBV core protein yields DN mutants that inhibit viral replication at the posttranslational level. To elucidate the mechanism(s) of their antiviral action, we analyzed the effect of the DN mutants on RNA pregenome packaging and nucleocapsid assembly. Core-Pol and core-S, but not core-lacZ or core-GFP, markedly interfered with RNA pregenome packaging. Nucleocapsid formation was not affected by any of the mutants. The DN core-GFP fusion protein formed mixed particles with wild-type core protein in the cytoplasm of cotransfected cells and interfered with reverse transcription of the viral pregenome. A subpopulation of chimeric nucleocapsids, however, was shown to overcome the block in DNA synthesis and produce mature viral DNA. Thus, at least 2 steps within the viral life cycle can be targeted by DN DHBV core proteins: 1) packaging of the viral pregenome; and 2) reverse transcription within mixed particles. The fact that some mixed particles retain replication competence demonstrates a high structural flexibility of nucleocapsids and indicates a possible mechanism of viral escape. PMID- 10385673 TI - An infectious molecular clone of a Japanese genotype 1b hepatitis C virus. AB - We describe an infectious molecular clone of a Japanese genotype 1b strain of hepatitis C virus (HCV-N). The molecularly cloned sequence of HCV-N was compared with alignments of other HCV sequences, leading to the identification of 15 unique, nonconservative amino acid substitutions within the HCV-N open reading frame (ORF). These were repaired to the consensus genotype 1b residue, and the infectivity of RNA transcribed from the repaired clone was assessed by intrahepatic inoculation of a chimpanzee. Viral RNA was first detected in the serum of this chimpanzee 3 weeks following inoculation, and was intermittently present over the next 14 weeks. A strong and persistent anti-HCV serological response developed 13 weeks following inoculation, with seroconversion in the recombinant immunoblot assay (RIBA). A weaker, transient serological response, characterized by seroconversion in a third-generation enzyme-linked immunosorbent assay (ELISA) but not RIBA, occurred between weeks 1 and 5. This may have represented an anamnestic response to HCV antigens translated directly from the intrahepatically inoculated RNA, because the animal previously had undergone 2 unsuccessful attempts at rescue of HCV by intrahepatic RNA inoculation. There was neither biochemical nor histological evidence of liver disease. Although this is within the range of expected outcomes in an HCV-naive chimpanzee, prior immunologic priming may have modified the infection in this animal. The HCV-N clone is the first infectious molecular clone of HCV that is comprised entirely of genotype 1b sequence, and it contains an ORF sequence that is significantly divergent from that of a previously described genotype 1a/1b chimera. PMID- 10385674 TI - Primary sclerosing cholangitis. PMID- 10385675 TI - Hepatocyte growth factor for liver disease. PMID- 10385676 TI - Eosinophils and primary biliary cirrhosis-stoking the fire? PMID- 10385677 TI - Prednisone withdrawal advocates: beware of recurrent autoimmune hepatitis. PMID- 10385678 TI - Selective substrates for non-neuronal monoamine transporters. AB - The recently identified transport proteins organic cation transporter 1 (OCT1), OCT2, and extraneuronal monoamine transporter (EMT) accept dopamine, noradrenaline, adrenaline, and 5-hydroxytryptamine as substrates and hence qualify as non-neuronal monoamine transporters. In the present study, selective transport substrates were identified that allow, by analogy to receptor agonists, functional discrimination of these transporters. To contrast efficiency of solute transport, stably transfected 293 cell lines, each expressing a single transporter, were examined side by side in uptake experiments with radiolabeled substrates. Normalized uptake rates indicate that tetraethylammonium, with a rate of about 0.5 relative to 1-methyl-4-phenylpyridinium (MPP+), is a good substrate for OCT1 and OCT2. It was not, however, accepted as substrate by EMT. Choline was transported exclusively by OCT1, with a rate of about 0.5 relative to MPP+. Histamine was a good substrate with a rate of about 0.6 relative to MPP+ for OCT2 and EMT, but was not transported by OCT1. Guanidine was an excellent substrate for OCT2, with a rate as high as that of MPP+. Transport of guanidine by OCT1 was low, and transport by EMT was negligible. With the guanidine derivatives cimetidine and creatinine, a pattern strikingly similar to guanidine was observed. Collectively, these substrates reveal key differences in solute recognition and turnover and thus challenge the concept of "polyspecific" organic cation transporters. In addition, our data, when compared with previous studies, suggest that OCT2 corresponds to the organic cation/H+ antiport mechanism in renal brush-border membrane vesicles, and that EMT corresponds to the guanidine/H+ antiport mechanism in membrane vesicles from placenta and intestine. PMID- 10385679 TI - Functional alpha6-containing nicotinic receptors are present in chick retina. AB - Despite the fact that the neuronal chick alpha6 subunit was first cloned several years ago and recently has been shown to form acetylcholine (ACh)-activated channels in heterologous systems, no information is yet available concerning the structure and function of the alpha6-containing nicotinic receptors in neuronal tissues. Using subunit-specific antibodies directed against two different epitopes of the chick alpha6 subunit, we performed immunoprecipitation experiments on immunopurified alpha6-containing receptors radiolabeled with the nicotinic agonist [3H]epibatidine (Epi): almost all of the alpha6 receptors contained the beta4 subunit, 51% the beta3 subunit, 42% the alpha3 subunit, and 7.5% the beta2 subunit. Western blot analyses of the purified receptors confirmed the presence of the alpha3, beta3, beta2, and beta4 subunits, and the absence of the alpha4, alpha5, and alpha7 subunits. The alpha6-containing receptors bind [3H]Epi (Kd = 35 pM) and a number of other nicotinic agonists with very high affinity, the rank order being Epi >> cytisine > nicotine > 1, 1-dimethyl-4 phenylpiperazinium > acetylcholine > carbamylcholine. The alpha6 receptors also have a distinct antagonist pharmacological profile with a rank order of potency of alpha-conotoxin MII > methyllycaconitine > dihydro-beta-erythroydine > MG624 > d-tubocurarine > decamethonium > hexamethonium. When reconstituted in lipid bilayers, the alpha6-containing receptors form functional cationic channels with a main conductance state of 48 pS. These channels are activated by nicotinic agonists in a dose-dependent manner, and blocked by the nicotinic antagonist d tubocurarine. PMID- 10385680 TI - DNA interactions of new antitumor aminophosphine platinum(II) complexes. AB - Mechanistic studies are presented of a novel class of aminophosphine platinum(II) complexes as potential anticancer agents. These new agents, which have demonstrated activity against murine and human tumor cells including those resistant to cisplatin are cis-[PtCl2(Me2N(CH2)3PPh2-P)2] (Com1) and cis [PtCl(C6H11NH(CH2)2PPh2-N,P)(C6H11NH(CH2) 2PPh2-P)] (Com2). We studied modifications of natural and synthetic DNAs in cell-free media by Com1 and Com2 by various biomedical and biophysical methods and compared the results with those obtained when DNA was modified by cisplatin. The results indicated that Com1 and Com2 coordinated to DNA faster than cisplatin. Bifunctional Com1 formed DNA adducts coordinating to single adenine or guanine residues or by forming cross links between these residues. In comparison with cisplatin, Com1 formed the adducts more frequently at adenine residues and also formed fewer bidentate lesions. The monofunctional Com2 only formed DNA monodentate adducts at guanine residues. In addition, Com1 terminated DNA synthesis in vitro more efficiently than cisplatin whereas Com2 blocked DNA synthesis only slightly. DNA unwinding studies, measurements of circular dichroism spectra, immunochemical analysis, and studies of the B-Z transition in DNA revealed conformational alterations induced by the adducts of Com1, which were distinctly different from those induced by cisplatin. Com2 had little influence on DNA conformation. It is suggested that the activity profile of aminophosphine platinum(II) complexes, which is different from that of cisplatin and related analogs, might be associated with the specific DNA binding properties of this new class of platinum(II) compounds. PMID- 10385681 TI - Characterization and implications of estrogenic down-regulation of human catechol O-methyltransferase gene transcription. AB - Catechol-O-methyltransferase (COMT, EC 2.1.1.6) is a ubiquitous enzyme that is crucial to the metabolism of carcinogenic catechols and catecholamines. Regulation of human COMT gene expression may be important in the pathophysiology of various human disorders including estrogen-induced cancers, Parkinson's disease, depression, and hypertension. The gender difference in human COMT activity and variations in rat COMT activity during the estrous cycle led us to explore whether estrogen can regulate human COMT gene transcription. Our Northern analyses showed that physiological concentrations of 17-beta-estradiol (10(-9) 10(-7) M) could decrease human 1. 3-kilobase COMT mRNA levels in MCF-7 cells in a time- and dose-dependent manner through an estrogen receptor-dependent mechanism. Two DNA fragments immediately 5' to the published human COMT gene proximal and distal promoters were cloned. Sequence analyses revealed several half-palindromic estrogen response elements and CCAAT/enhancer binding protein sites. By cotransfecting COMT promoter-chloramphenicol acetyltransferase reporter genes with human estrogen receptor cDNA and pSV-beta-galactosidase plasmids into COS-7 cells, we showed that 17-beta-estradiol could down-regulate chloramphenicol acetyltransferase activities, and COMT promoter activities dose-dependently. Functional deletion analyses of COMT promoters also showed that this estrogenic effect was mediated by a 280 base pair fragment with two putative half palindromic estrogen response elements in the proximal promoter and a 323-base pair fragment with two putative CCAAT/enhancer binding protein sites in the distal promoter. Our findings provide the first evidence and molecular mechanism for estrogen to inhibit COMT gene transcription, which may shed new insight into the role of estrogen in the pathophysiology of different human disorders. PMID- 10385682 TI - Inhibition of calcium/calmodulin-dependent protein kinase II in rat hippocampus attenuates morphine tolerance and dependence. AB - Learning and memory have been suggested to be important in the development of opiate addiction. Based on the recent findings that calcium/calmodulin-dependent protein kinase II (CaMKII) is essential in learning and memory processes, and morphine treatment increases CaMKII activity in hippocampus, the present study was undertaken to examine whether inhibition of hippocampal CaMKII prevents morphine tolerance and dependence. Here, we report that inhibition of CaMKII by intrahippocampal dentate gyrus administration of the specific inhibitors KN-62 and KN-93 to rats significantly attenuated the tolerance to the analgesic effect of morphine and the abstinence syndrome precipitated by opiate antagonist naloxone. In contrast, both KN-04 and KN-92, the inactive structural analogs of KN-62 and KN-93, failed to attenuate morphine tolerance and dependence, indicating that the observed effects of KN-62 and KN-93 are mediated through inhibition of CaMKII. Furthermore, administration of CaMKII antisense oligonucleotide into rat hippocampal dentate gyrus, which decreased the expression of CaMKII specifically, also attenuated morphine tolerance and dependence, while the corresponding sense oligonucleotide of CaMKII did not exhibit such inhibitory effect. Moreover, the KN-62 treatment abolished the rewarding properties of morphine as measured by the conditioned place preference. These results suggest that hippocampal CaMKII is critically involved in the development of morphine tolerance and dependence, and inhibition of this kinase may have some therapeutic benefit in the treatment of opiate tolerance and dependence. PMID- 10385683 TI - Mimicry in primary rat hepatocyte cultures of the in vivo perivenous induction by phenobarbital of cytochrome P-450 2B1 mRNA: role of epidermal growth factor and perivenous oxygen tension. AB - Treatment of male rats with phenobarbital (PB) results in a perivenous and mid zonal pattern of cytochrome P-450 (CYP)2B1 mRNA expression within the liver acinus. The mechanism of this zonated induction is still poorly understood. In this study sinusoidal gradients of oxygen and epidermal growth factor (EGF) besides those of the pituitary-dependent hormones growth hormone (GH), thyroxine (T4), and triiodothyronine (T3) were considered to be possible determinants for the zonated induction of the CYP2B1 gene in liver. Moreover, heme proteins seem to play a key role in oxygen sensing. Therefore, the influence of arterial (16% O2) and venous (8% O2) oxygen tension (pO2), and of the heme synthesis inhibitors CoCl2 and desferrioxamine (DSF) on PB-dependent CYP2B1 mRNA induction as well as the repression by EGF and, for comparison, by GH, T4, and T3, of the induction under arterial and venous pO2 were investigated in primary rat hepatocytes. Within 3 days, phenobarbital induced CYP2B1 mRNA to maximal levels under arterial pO2 and to about 40% of maximal levels under venous pO2. CoCl2 annihilated induction by PB under both oxygen tensions, whereas desferrioxamine and heme abolished the positive modulation by O2, suggesting that heme is a necessary component for O2 sensing. EGF suppressed CYP2B1 mRNA induction by PB only under arterial but not under venous pO2, whereas GH, T4, and T3 inhibited induction under both arterial and venous pO2. Thus, in hepatocyte cultures, an O2 gradient in conjunction with EGF mimicked the perivenous induction by PB of the CYP2B1 gene observed in the liver in vivo. PMID- 10385684 TI - Humanization of mouse 5-hydroxytryptamine1B receptor gene by homologous recombination: in vitro and in vivo characterization. AB - We replaced the coding region of the murine 5-hydroxytryptamine (5-HT)1B receptor by the human 5-HT1B receptor using homologous recombination in embryonic stem cells and generated and characterized homozygous transgenic mice that express only the human (h) 5-HT1B receptor. The distribution patterns of h5-HT1B and murine (m) 5-HT1B receptor mRNA and binding sites in brain sections of transgenic and wild-type mice were identical as measured by in situ hybridization histochemistry and radioligand receptor autoradiography. When measured in parallel under identical conditions, the h5-HT1B receptor expressed in mouse brain had the same pharmacological characteristics as that in human brain. Stimulation by 5-HT1B agonists of [35S]guanosine-5'-O-(3-thio)triphosphate binding in brain sections demonstrated the functional coupling of the h5-HT1B receptor to G proteins in mouse brain. In tissue slices from various brain regions, electrically stimulated [3H]5-HT release was not modified by 5-HT1B agonists in tissue from either transgenic and wild-type mice; a 5-HT1B antagonist enhanced electrically stimulated [3H]5-HT release in wild-type mouse brain, but was ineffective in the transgenics. The centrally active 5-HT1A/5-HT1B agonist RU24969 induced hypothermia but did not increase locomotor activity in the transgenic mice. The ineffectiveness of RU24969 in the transgenic mice could be due to the lower affinity of the compound for the h5-HT1B receptor compared with the m5-HT1B receptor. The present study demonstrates a complete replacement of the mouse receptor by its human receptor homolog and a functional coupling to G proteins. However, modulation of [3H]5-HT release could not be shown. Furthermore, behavioral effects were not clearly observed, which may be due to a lack of appropriate tools. PMID- 10385685 TI - Pattern of mutations that results in loss of reduced folate carrier function under antifolate selective pressure augmented by chemical mutagenesis. AB - Chemical mutagenesis with N-methyl-N-nitrosourea was employed to study the pattern of mutations in the reduced folate carrier (RFC1) that results in transport-related methotrexate resistance and to identify amino acid residues that are critical to carrier structure and/or function. Thirty-four methotrexate transport-defective L1210 leukemia cell lines were isolated with folic acid as the sole folate source under antifolate selective pressure. The RFC1 mRNA levels were comparable with, or not substantially decreased, in most of these cell lines relative to wild-type L1210 cells. The molecular basis for the transport defects was investigated by sequencing multiple RFC1 cDNA clones isolated from these mutants by reverse transcription-polymerase chain reaction, which encompassed the entire coding region. The mutations identified were further confirmed either by direct sequencing or, when applicable, by restriction analysis of total reverse transcription-polymerase chain reaction products. The majority of mutations (21) led to single amino acid substitutions that were in, or near, 9 of 12 predicted transmembrane domains, with the highest frequencies in the first, fifth, and eighth. There were no mutations in the sixth, ninth, and twelfth transmembrane domains. Glycine, serine, and arginine were the most frequently mutated residues. These data suggest that several transmembrane domains, rather than the amino- and carboxyl-termini, and the large intracellular loop between the sixth and seventh transmembrane domains play key roles as sites for RFC1 inactivation because of single point mutations. This panel of mutated cell lines offers an important resource for studies on RFC1 structure-function and for the evaluation of transport-related cross-resistance patterns with new-generation antifolate inhibitors of tetrahydrofolate cofactor-dependent enzymes. PMID- 10385686 TI - Topoisomerase poisoning activity of novel disaccharide anthracyclines. AB - Doxorubicin and idarubicin are very effective anticancer drugs in the treatment of human hematological malignancies and solid tumors. These agents are well known topoisomerase II poisons; however, some anthracycline analogs recently have been shown to poison topoisomerase I. In the present work, we assayed novel disaccharide analogs and the parent drug, idarubicin, for their poisoning effects of human topoisomerase I and topoisomerases IIalpha and IIbeta. Drugs were evaluated with a DNA cleavage assay in vitro and with a yeast system to test whether the agents were able to poison the enzymes in vivo. We have found that the test agents are potent poisons of both topoisomerases IIalpha and IIbeta. The axial orientation of the second sugar relative to the first one of the novel disaccharide analogs was shown to be required for poisoning activity and cytotoxicity. Interestingly, idarubicin and the new analogs stimulated topoisomerase I-mediated DNA cleavage at low levels in vitro. As expected, the cytotoxic level of the drug was highly affected by the content of topoisomerase II; nevertheless, the test agents had a yeast cell-killing activity that also was weakly dependent on cellular topoisomerase I content. The results are relevant for the full understanding of the molecular mechanism of topoisomerase poisoning by anticancer drugs, and they define structural determinants of anthracyclines that may help in the rational design of new compounds directed against topoisomerase I. PMID- 10385687 TI - Ethanol inhibition of synaptically evoked kainate responses in rat hippocampal CA3 pyramidal neurons. AB - Many studies have demonstrated that intoxicating concentrations of ethanol (10 100 mM) can selectively inhibit the component of glutamatergic synaptic transmission mediated by N-methyl-D-aspartate (NMDA) receptors while having little or no effect on excitatory synaptic transmission mediated by non-NMDA receptors [i.e., alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) and/or kainate (KA) receptors]. However, until the recent development of highly selective AMPA receptor antagonists, it was not possible to assess the relative contribution of AMPA and KA receptors to non-NMDA receptor-mediated synaptic transmission or to determine whether these glutamate receptor subtypes differed in their sensitivity to ethanol. In the present experiments, we used the highly selective AMPA receptor antagonist LY 303070 to pharmacologically isolate KA receptor-mediated excitatory postsynaptic currents (EPSCs) in rat hippocampal CA3 pyramidal neurons and tested their sensitivity to ethanol. Concentrations of ethanol as low as 20 mM significantly and reversibly depressed KA EPSCs. Ethanol also inhibited KA currents evoked by direct pressure application of KA in the presence of LY 303070, suggesting that this inhibition was mediated by a postsynaptic action. In contrast, ethanol had no effect on AMPA EPSCs in these cells, even at the highest concentration tested (80 mM). Ethanol significantly inhibited NMDA EPSCs in these neurons, but these responses were less sensitive to ethanol than KA EPSCs. These results suggest that in addition to its well described depressant effect on NMDA receptor-mediated synaptic transmission, ethanol has an even greater inhibitory effect on glutamatergic synaptic transmission mediated by KA receptors in rat hippocampal CA3 pyramidal neurons. PMID- 10385688 TI - Endrin inhibits adipocyte differentiation by selectively altering expression pattern of CCAAT/enhancer binding protein-alpha in 3T3-L1 cells. AB - The effects of selected chlorinated cyclodiene pesticides on the adipocyte differentiation process were examined using the 3T3-L1 adipocyte model in vitro. Endrin was found to cause a dose-dependent inhibition of adipocyte differentiation in 3T3-L1 cells. Aldrin and dieldrin were less potent than endrin in interfering with the adipogenic process. Endrin's inhibitory effect was effective only when the pesticide was present in the medium during the first 48 h after exposure of 3T3-L1 cells to adipogenic inducers. Immunoblots analysis revealed that endrin caused a dose-dependent, selective inhibition of the intracellular levels of CCAAT enhancer binding protein (C/EBP)alpha without altering the expression patterns of C/EBPbeta or C/EBPdelta along the differentiation. Supershift analysis showed that DNA-binding capacity of C/EBPalpha was affected most by endrin treatment. Endrin also caused a decrease in the elevation of the adipogenic factor peroxisome proliferator-activated receptor (PPAR)gamma elicited by the adipogenic inducers. However, the cotreatment with troglitazone, a thiazolidinedione known to activate PPARgamma, did not suppress the antiadipogenic action of endrin, indicating that its direct action site is not PPARgamma receptor. Endrin also altered the pattern of activation of nuclear factor-kappaB, a factor activated by 12-O tetradecanoylphorbol-13-acetate and tumor necrosis factor-alpha, which are known to interfere with adipocyte differentiation. Thus, endrin inhibited the normal decrease in nuclear factor-kappaB-DNA binding observed as cells are acquiring the adipocyte phenotype at a late stage of differentiation. Our results suggest that endrin inhibits adipocyte differentiation through the specific suppression of C/EBPalpha. PMID- 10385689 TI - Dithiothreitol enhances arsenic trioxide-induced apoptosis in NB4 cells. AB - Recently, arsenic trioxide (As2O3) was reported to induce clinical remission in patients with acute promyelocytic leukemia. Modulation of protein phosphorylation by binding to the vicinal thiols has been suggested as a possible mechanism. We found that phenylarsine oxide, a strong vicinal thiol-binding agent, neither induced nuclear fragmentation or DNA laddering nor increased caspase activity in NB4 cells; however, As2O3 and a weak thiol-binding agent, dimethylarsinic acid, did increase activity. Dithiothreitol (DTT) effectively suppressed the phenylarsine oxide-inhibited cellular reductive capacity, but unexpectedly, enhanced As2O3-induced apoptosis in NB4 cells. As2O3-induced and As2O3-plus-DTT induced apoptosis in NB4 cells was modulated by oxidant modifiers, but not by nitric oxide synthase inhibitors. These results demonstrate that DTT, a dithiol agent and known antidote for trivalent inorganic arsenic, enhances the toxicity of As2O3, thereby opening a new research direction for the mechanisms of arsenic toxicity and perhaps also helping in the development of new therapeutic strategies for treating leukemias. PMID- 10385690 TI - Preferential coassembly of alpha4 and delta subunits of the gamma-aminobutyric acidA receptor in rat thalamus. AB - Pharmacological study of rat thalamic gamma-aminobutyric acidA (GABAA) receptors revealed the presence of two distinct populations, namely, diazepam-sensitive and diazepam-insensitive [3H]Ro15-4513 binding sites accounting for 94 +/- 2% (1339 +/- 253 fmol/mg protein) and 6 +/- 2% (90 +/- 44 fmol/mg protein) of total sites, respectively. Thalamic diazepam-insensitive sites exhibited a pharmacology that was distinct from diazepam-sensitive sites but comparable to that of the alpha4beta3gamma2 subtype of the GABAA receptor stably expressed in L(tk-) cells. Immunoprecipitation experiments with a specific anti-alpha4-antiserum immunoprecipitated 20 and 7% of total thalamic [3H]muscimol and [3H]Ro15-4513 sites, respectively. Combinatorial immunoprecipitation using antisera against the alpha4, gamma2, and delta subunit revealed that alpha4delta- and alpha4gamma2 containing receptors account for 13 +/- 2 and 8 +/- 3% of [3H]muscimol sites from thalamus, respectively. It also indicated that all delta subunits coexist with an alpha4 subunit in this brain region. In conclusion, our results show that in rat thalamus both alpha4betagamma2 and alpha4betadelta subtypes are expressed but alpha4betadelta is the major alpha4-containing GABAA receptor population. PMID- 10385691 TI - Modulation of expression of endothelial nitric oxide synthase by nordihydroguaiaretic acid, a phenolic antioxidant in cultured endothelial cells. AB - Retrospective epidemiological studies have suggested that antioxidant therapy may decrease cardiovascular morbidity and mortality rates, although the mechanisms for this effect remain unclear. In the present study, we demonstrate that selective antioxidants can enhance expression of endothelial nitric oxide synthase (eNOS). We found that the antioxidants nordihydroguaiaretic acid (NDGA), catechol, glutaryl probucol, and N-acetylcysteine increased eNOS expression in cultured bovine aortic endothelial cells (BAECs). NDGA seemed to be the most potent of the phenolic antioxidants, producing a 3-fold increase in eNOS mRNA. This effect of NDGA was enhanced by nonphenolic antioxidants such as N acetylcysteine and ascorbic acid. Nuclear run-on studies indicated that NDGA increased eNOS transcription. A similar increase in eNOS protein content was observed with Western blot analysis after treating BAECs or human aortic endothelial cells with NDGA. Exposure of BAECs to NDGA enhanced NO production, as measured by electron paramagnetic resonance spin trapping and eNOS activity, as measured by [14C]arginine-to-[14C]citrulline assay. Methylation of the phenolic hydroxyl groups completely inhibited the NDGA effect on eNOS mRNA levels. This effect of NDGA was not due to inhibition of lipoxygenase because cis-5,8,11,14 eicosatetraynoic acid did not alter eNOS expression. We conclude that antioxidants may not only increase the bioactivity of nitric oxide but also enhance expression of the eNOS enzyme. Such an effect may prove useful in conditions such as hypertension and atherosclerosis, in which nitric oxide production and/or biological activity is impaired. PMID- 10385692 TI - Inhibition of cyclic GMP-binding cyclic GMP-specific phosphodiesterase (Type 5) by sildenafil and related compounds. AB - The cGMP-binding cGMP-specific phosphodiesterase (PDE5) degrades cGMP and regulates the intracellular level of cGMP in many tissues, including the smooth muscle of the corpus cavernosum of the penis. Sildenafil (Viagra), a specific PDE5 inhibitor, promotes penile erection by blocking the activity of PDE5, which causes cGMP to accumulate in the corpus cavernosum. In the present study, sildenafil, like other PDE5 inhibitors, stimulates cGMP binding to the allosteric sites of PDE5 by interacting at the catalytic site of this enzyme, but the drug does not compete with cGMP for binding at the allosteric sites. Both sildenafil and zaprinast are competitive inhibitors of PDE5, and double-inhibition analysis shows that these two inhibitors added together interact with the catalytic site of PDE5 in a mutually exclusive manner. After site-directed mutagenesis of each of 23 conserved amino acid residues in the catalytic domain of PDE5, the pattern of changes in the IC50 values for sildenafil or UK-122764 is similar to that found for zaprinast. However, among the three inhibitors, sildenafil exhibits the most similar pattern of changes in the IC50 to that found for the affinity of cGMP, implying similar interactions with the catalytic domain. This may explain in part the stronger inhibitory potency of sildenafil for wild-type PDE5 compared with the other inhibitors [sildenafil (Ki = 1 nM) > UK-122764 (Ki = 5 nM) > zaprinast (Ki = 130 nM)]. The affinity of each of these inhibitors for PDE5 is much higher than that of cGMP itself (Km = 2000 nM). It is concluded that residues such as Tyr602, His607, His643, and Asp754 may form important interactions for sildenafil in PDE5, but because these amino acids are conserved in all mammalian PDEs, the selectivity and potency of sildenafil is likely to be provided by a nonconserved residue or residues in the PDE5 catalytic domain. PMID- 10385693 TI - Farnesol and geranylgeraniol prevent activation of caspases by aminobisphosphonates: biochemical evidence for two distinct pharmacological classes of bisphosphonate drugs. AB - Recently, advances have been made in understanding the molecular mechanisms by which bisphosphonate drugs inhibit bone resorption. Studies with the macrophage like cell line J774 have suggested that alendronate, an amino-containing bisphosphonate, causes apoptosis by preventing post-translational modification of GTP-binding proteins with isoprenoid lipids. However, clodronate, a nonaminobisphosphonate, does not inhibit protein isoprenylation but can be metabolized intracellularly to a cytotoxic, beta-gamma-methylene (AppCp-type) analog of ATP. These observations raise the possibility that bisphosphonates can be divided into two groups with distinct molecular mechanisms of action depending on the nature of the R2 side chain. We addressed this question by directly comparing the ability of three aminobisphosphonates (alendronate, ibandronate, and pamidronate) and three nonaminobisphosphonates (clodronate, etidronate, and tiludronate) to inhibit protein isoprenylation and activate caspase-3-like proteases or to be metabolized to AppCp-type nucleotides by J774 cells. All three aminobisphosphonates inhibited protein isoprenylation and activated caspase-3 like proteases. Apoptosis and caspase activation after 24-h treatment with the aminobisphosphonates could be prevented by addition of farnesol or geranylgeraniol, confirming that these bisphosphonates inhibit the metabolic mevalonate pathway. No AppCp-type metabolites of the aminobisphosphonates could be detected by mass spectrometry. The three nonaminobisphosphonates did not inhibit protein isoprenylation or cause activation of caspase-3-like proteases, but were incorporated into AppCp-type nucleotides. Taken together, these observations clearly demonstrate that bisphosphonate drugs can be divided into two pharmacological classes: the aminobisphosphonates, which act by inhibiting protein isoprenylation, and the less potent nonaminobisphosphonates, which act through the intracellular accumulation of AppCp-type metabolites. PMID- 10385694 TI - Cross-resistance to ionizing radiation in a murine leukemic cell line resistant to cis-dichlorodiammineplatinum(II): role of Ku autoantigen. AB - cis-Dichlorodiammineplatinum(II) (CDDP; cisplatin) is commonly used in combination with ionizing radiation (IR) in the treatment of various malignancies. In vitro, many observations suggest that acquisition of CDDP resistance in cell lines confers cross-resistance to IR, but the molecular mechanisms involved have not been well documented yet. We report here the selection and characterization of a murine CDDP-resistant L1210 cell line (L1210/3R) that exhibits cross-resistance to IR because of an increased capacity to repair double-strand breaks compared with parental cells (L1210/P). In resistant cells, electrophoretic mobility shift assays revealed an increased DNA end binding activity that could be ascribed, by supershifting the retardation complexes with antibodies, to the autoantigen Ku. The heterodimeric Ku protein, composed of 86-kDa (Ku80) and 70-kDa (Ku70) subunits, is the DNA-targeting component of DNA-dependent protein kinase (DNA-PK), which plays a critical role in mammalian DNA double-strand breaks repair. The increased Ku-binding activity in resistant cells was associated with an overexpression affecting specifically the Ku80 subunit. These data strongly suggest that the increase in Ku activity is responsible for the phenotype of cross-resistance to IR. In addition, these observations, along with previous results from DNA-PK- mutant cells, provide evidence in favor of a role of Ku/DNA-PK in resistance to CDDP. These results suggest that Ku activity may be an important molecular target in cancer therapy at the crossroad between cellular responses to CDDP and IR. PMID- 10385695 TI - Induction of alkylator (melphalan) resistance in HL60 cells is accompanied by increased levels of topoisomerase II expression and function. AB - Human leukemic HL60 cells were selected for resistance to alkylating agents by stepwise exposure to increasing concentrations of L-phenylalanine mustard (melphalan). The resulting resistant cell line (R-HL60) was 4-fold resistant (melphalan IC50 value, 27.84 +/- 4.2 microM) to melphalan compared with parental HL60 cells (melphalan IC50 value, 6.9 +/- 1.78 microM). Nuclear extracts from R HL60 cells possess a approximately 4-fold increase in DNA topoisomerase II activity compared with parental HL60 cells. As determined using Western blot analysis, the level of topoisomerase IIalpha protein expressed in R-HL60 cells was approximately 3-fold that of parental HL60 cells. However, there were no differences observed in the level of topoisomerase IIbeta protein, in the topoisomerase I activity, or in the level of topoisomerase I protein expression comparing the two cell lines. R-HL60 cells were 5-fold more sensitive than parental HL60 cells to the cytotoxic effect of the topoisomerase II inhibitor doxorubicin. The sensitivity to the cytotoxic effects of the topoisomerase I inhibitor camptothecin did not differ in R-HL60 and parental HL60 cell lines. Preincubation with doxorubicin significantly increased melphalan-induced interstrand DNA cross-link formation and cytotoxicity in R-HL60 cells compared with the parental HL60 cells. The affinity of topoisomerase II for UV-irradiated cross-linked HL60 DNA was increased by approximately 2.5-fold compared with that of HL60 native DNA. The affinity of topoisomerase II for both UV-irradiated (cross-linked) and native DNA was significantly decreased after doxorubicin pretreatment. Elevated topoisomerase II activity and the increased affinity of topoisomerase II for cross-linked DNA in melphalan-resistant cells seems to contribute to alkylator resistance by changing DNA topology, thereby facilitating DNA repair. PMID- 10385696 TI - Abnormal regulation of the sympathetic nervous system in alpha2A-adrenergic receptor knockout mice. AB - alpha2-Adrenergic receptors (ARs) play a key role in regulating neurotransmitter release in the central and peripheral sympathetic nervous systems. To date, three subtypes of alpha2-ARs have been cloned (alpha2A, alpha2B, and alpha2C). Here we describe the physiological consequences of disrupting the gene for the alpha2A AR. Mice lacking functional alpha2A subtypes were compared with wild-type (WT) mice, with animals lacking the alpha2B or alpha2C subtypes, and with mice carrying a point mutation in the alpha2A-AR gene (alpha2AD79N). Deletion of the alpha2A subtype led to an increase in sympathetic activity with resting tachycardia (knockout, 581 +/- 21 min-1; WT, 395 +/- 21 min-1), depletion of cardiac tissue norepinephrine concentration (knockout, 676 +/- 31 pg/mg protein; WT, 1178 +/- 98 pg/mg protein), and down-regulation of cardiac beta-ARs (Bmax: knockout, 23 +/- 1 fmol/mg protein; WT, 31 +/- 2 fmol/mg protein). The hypotensive effect of alpha2 agonists was completely absent in alpha2A-deficient mice. Presynaptic alpha2-AR function was tested in two isolated vas deferens preparations. The nonsubtype-selective alpha2 agonist dexmedetomidine completely blocked the contractile response to electrical stimulation in vas deferens from alpha2B-AR knockout, alpha2C-AR knockout, alpha2AD79N mutant, and WT mice. The maximal inhibition of vas deferens contraction by the alpha2 agonist in alpha2A AR knockout mice was only 42 +/- 9%. [3H]Norepinephrine release studies performed in vas deferens confirmed these findings. The results indicate that the alpha2A AR is a major presynaptic receptor subtype regulating norepinephrine release from sympathetic nerves; however, the residual alpha2-mediated effect in the alpha2A AR knockout mice suggests that a second alpha2 subtype (alpha2B or alpha2C) also functions as a presynaptic autoreceptor to inhibit transmitter release. PMID- 10385698 TI - Phosphodiesterase 4B2 is the predominant phosphodiesterase species and undergoes differential regulation of gene expression in human monocytes and neutrophils. AB - The type 4 phosphodiesterase (PDE4) is the predominant PDE isozyme in various leukocytes and plays a key role in the regulation of inflammatory cell activation. There are four PDE4 subtypes (A, B, C, and D), and within each subtype, there are multiple variants. Very recently, we found in monocytes that PDE4B gene expression is selectively induced by lipopolysaccharide (LPS) and that the induction is inhibited by interleukin (IL)-10 and IL-4. In this study, we show that the PDE4B gene is constitutively expressed in neutrophils and that this expression remains unaffected by LPS or IL-10. PDE4B is the predominant subtype in neutrophils and in unstimulated or LPS-stimulated monocytes, and in these cells, the PDE4B2 variant is the only detectable molecular species of PDE4B. Therefore, PDE4B2 is the predominant PDE isoform in human neutrophils and monocytes, and its expression is regulated differently by these two cell types. Furthermore, leukocytes are the most dominant source of PDE4B2, suggesting that PDE4B2 is a relatively specific target for discovering anti-inflammatory drugs. PMID- 10385697 TI - Both inducible and constitutive activator protein-1-like transcription factors are used for transcriptional activation of the galanin gene by different first and second messenger pathways. AB - We investigated trans-acting factors mediating galanin (GAL) gene activation by protein kinase-dependent signal transduction pathways in chromaffin cells. GAL mRNA up-regulation via the protein kinase A (PKA) pathway (25 microM forskolin) required new protein synthesis. Stimulation via protein kinase C (0.1 microM phorbol myristate acetate) did not. The involvement of activator protein-1(AP-1) and cAMP response element-binding protein (CREB) in serine/threonine protein kinase activation of GAL gene transcription was assessed. Cotransfection of a GAL reporter gene along with expression plasmids encoding c-Jun plus c-Fos, or the catalytic subunit of PKA (PKAbeta), resulted in a 4- to 8-fold enhancement of GAL reporter gene transcription. Transcriptional activation required the galanin 12-O tetradecanoylphorbol-13-acetate (phorbol-12-myristate-13-acetate) response element (GTRE) octamer sequence (TGACGCGG) in the proximal enhancer of the GAL gene, previously shown to confer phorbol ester responsiveness in chromaffin cells. CREB coexpression did not stimulate GAL gene transcription or increase transcriptional activation by PKAbeta. The GTRE preferentially bound in vitro synthesized Jun and Fos-Jun, compared with CREB, in electrophoretic mobility shift assays. The GTRE preference for binding AP-1-immunoreactive protein compared with CREB was even more pronounced in chromaffin cell nuclear extracts, in which the majority of GTRE-bound protein in electrophoretic mobility shift assays was supershifted with anti-Fos and anti-Jun antibodies. Thus, GAL gene regulation mediated by protein kinase activation appears to involve both constitutively expressed and inducible AP-1-related proteins. Elevated potassium stimulation of GAL mRNA was completely blocked, but pituitary adenylyl cyclase activating polypeptide and histamine stimulations were only partially blocked, by cycloheximide. Both inducible and constitutive pathways are therefore used by physiologically relevant first messengers that stimulate GAL biosynthesis in vivo. PMID- 10385699 TI - Mutation of a highly conserved aspartic acid in the beta2 adrenergic receptor: constitutive activation, structural instability, and conformational rearrangement of transmembrane segment 6. AB - Movements of transmembrane segments (TMs) 3 and 6 play a key role in activation of G protein-coupled receptors. However, the underlying molecular processes that govern these movements, and accordingly control receptor activation, remain unclear. To elucidate the importance of the conserved aspartic acid (Asp-130) in the Asp-Arg-Tyr motif of the beta2 adrenergic receptor (beta2AR), we mutated this residue to asparagine (D130N) to mimic its protonated state, and to alanine (D130A) to fully remove the functionality of the side chain. Both mutants displayed evidence of constitutive receptor activation. In COS-7 cells expressing either D130N or D130A, basal levels of cAMP accumulation were clearly elevated compared with cells expressing the wild-type beta2AR. Incubation of COS-7 cell membranes or purified receptor at 37 degrees C revealed also a marked structural instability of both mutant receptors, suggesting that stabilizing intramolecular constraints had been disrupted. Moreover, we obtained evidence for a conformational rearrangement by mutation of Asp-130. In D130N, a cysteine in TM 6, Cys-285, which is not accessible in the wild-type beta2AR, became accessible to methanethiosulfonate ethylammonium, a charged, sulfhydryl-reactive reagent. This is consistent with a counterclockwise rotation or tilting of TM 6 and provides for the first time structural evidence linking charge-neutralizing mutations of the aspartic acid in the DRY motif to the overall conformational state of the receptor. We propose that protonation of the aspartic acid leads to release of constraining intramolecular interactions, resulting in movements of TM 6 and, thus, conversion of the receptor to the active state. PMID- 10385700 TI - The staurosporine-like compound L-753,000 (NB-506) potentiates the neurotrophic effects of neurotrophin-3 by acting selectively at the TrkA receptor. AB - K-252b, a member of the staurosporine family of protein kinase inhibitors, selectively potentiates the activation of the nerve growth factor receptor, TrkA, by a nonpreferred ligand, neurotrophin-3 (NT-3), in a variety of cell types. At higher (micromolar) concentrations of K-252b, an inhibitory effect occurs because of the inhibitory action of K-252b on the Trk kinase. By examining analogs of K 252b, we identified the compound L-753,000 (NB-506), which potentiates the action of NT-3 on TrkA but is devoid of the inhibitory action of K-252b. L-753,000 was effective at nanomolar concentrations in a Chinese hamster ovary cell line that expressed TrkA but was devoid of p75, the low-affinity neurotrophin receptor. L 753,000 also potentiated the activation of mitogen-activating protein kinase signaling (downstream from Trk activation) by NT-3 in this cell line. Although L 753,000, like K-252b, had a negligible effect in the absence of NT-3, the compound was found to potentiate NT-3-induced survival in both rat and chick primary cultures of dissociated dorsal root ganglia (DRG) and on neurite outgrowth of chick DRG explants. Unlike K-252b, which at micromolar concentrations inhibits the survival response of NT-3 in dissociated rat DRG, L 753,000 continued to potentiate the actions of NT-3 up to a concentration of 10 microM. Furthermore, the compound, unlike K-252b, did not inhibit an unrelated protein kinase, protein kinase C, at concentrations up to 10 microM. Because L 753, 000 selectively potentiates the NT-3-induced stimulation of TrkA without inhibiting Trks and other protein kinases, it represents a novel class of selective modifiers of neurotrophin actions. PMID- 10385701 TI - The inhibition of mammalian 15-lipoxygenases by the anti-inflammatory drug ebselen: dual-type mechanism involving covalent linkage and alteration of the iron ligand sphere. AB - Mammalian lipoxygenases have been implicated in inflammation and atherosclerosis and, thus, lipoxygenase inhibitors may be of pharmacological interest. In cells, lipoxygenases occur in a catalytically silent ground state that requires activation to become active. We found that the seleno-organic drug ebselen [2 phenyl-1, 2-benzisoselenazol-3(2H)-one], which exhibits anti-inflammatory properties, irreversibly inhibited pure rabbit 15-lipoxygenase, with an IC50 in the nM range when preincubated with the enzyme in the absence of fatty acid substrates. Subsequent dialysis, gel filtration, or substrate addition did not restore the enzyme activity, and experiments with [14C]ebselen indicated a covalent linkage of the drug. The presence of sulfhydryl compounds in the incubation mixture prevented both enzyme labeling and inactivation, but we did not see any reactivation when sulfhydryl compounds were added afterward. X-ray absorption studies indicated that ebselen did alter the geometry of the iron ligand sphere, and the data are consistent with an iron complexation by the drug. When fatty acid substrate was present during lipoxygenase-ebselen interaction, the inhibitory potency was strongly reduced and a competitive mode of action was observed. These data suggest that ebselen inactivated the catalytically silent ground-state lipoxygenase irreversibly by covalent linkage and alteration of the iron ligand sphere. In contrast, it functions as a competitive inhibitor of the catalytically active enzyme species. The pharmacological relevance of ebselen as a potential in vivo lipoxygenase inhibitor will be discussed. PMID- 10385702 TI - Modulation of fibroblast growth factor-2 receptor binding, signaling, and mitogenic activity by heparin-mimicking polysulfonated compounds. AB - Basic fibroblast growth factor (FGF-2) interacts with high-affinity tyrosine kinase fibroblast growth factor receptors (FGFRs) and low-affinity heparan sulfate proteoglycans (HSPGs) in target cells. Both interactions are required for FGF-2-mediated biological responses. Here we report the FGF-2 antagonist activity of novel synthetic sulfonic acid polymers with distinct chemical structures and molecular masses (MMs). PAMPS [poly(2-acrylamido-2-methyl-1-propanesulfonic acid)], (MM approximately 7,000-10,000), PAS [poly(anetholesulfonic acid)], (MM approximately 9,000-11,000), PSS [poly(4-styrenesulfonic acid)], (MM = 70,000), and poly(vinylsulfonic acid) (MM = 2,000), inhibited FGF-2 binding to HSPGs and FGFRs in fetal bovine aortic endothelial GM 7373 cells. They also abrogated the formation of the HSPG/FGF-2/FGFR ternary complex, as evidenced by their capacity to prevent FGF-2-mediated cell-cell attachment of FGFR-1-overexpressing, HSPG deficient Chinese hamster ovary cells to wild-type HSPG-bearing cells. Direct interaction of the polysulfonates with FGF-2 was demonstrated by their ability to protect the growth factor from proteolytic cleavage. Accordingly, molecular modeling, based on the crystal structure of the interaction of FGF-2 with a heparin hexamer, showed the feasibility of docking PAMPS into the heparin-binding domain of FGF-2. In agreement with their FGF-2-binding capacity, PSS, PAS, and PAMPS inhibited FGF-2-induced cell proliferation in GM 7373 cells and murine brain microvascular endothelial cells. The antiproliferative activity of these compounds was associated with the abrogation of FGF-2-induced tyrosine phosphorylation of FGFR-1. Moreover, the polysulfonates PSS and PAS inhibited FGF 2-induced activation of mitogen-activated protein kinase-1/2, involved in FGF-2 signal transduction. In conclusion, sulfonic acid polymers bind FGF-2 by mimicking heparin interaction. These compounds may provide a tool to inhibit FGF 2-induced endothelial cell proliferation in angiogenesis and tumor growth. PMID- 10385703 TI - Activation mechanism of human oxytocin receptor: a combined study of experimental and computer-simulated mutagenesis. AB - The aim of this study was to investigate the molecular changes associated with the transition of the human oxytocin receptor from its inactive to its active states. Mutation of the conserved arginine of the glutamate/aspartate-arginine tyrosine motif located in the second intracellular domain gave rise to the first known constitutively active oxytocin receptor (R137A), whereas mutation of the aspartic acid located in the second transmembrane domain led to an inactive receptor (D85A). The structural features of the constitutively active and inactive receptor mutants were compared with those of the wild type in its free and agonist-bound states. The results suggest that, although differently triggered, the activation process induced by the agonist and the activating mutation are characterized by the opening of a solvent exposed site formed by the 2nd intracellular loop, the cytosolic extension of helix 5, and the 3rd intracellular loop; on the contrary, the D85A mutation prevents oxytocin from triggering the opening of a cytosolic site. On the basis of these findings, we hypothesize that this cytosolic crevice plays an important role in G protein recognition. Finally, comparative analysis of the free- and agonist-bound forms of the wild-type oxytocin receptor and alpha1B adrenergic receptor suggests that the highly conserved polar amino acids and the seven helices play similar mechanistic roles in the different G protein-coupled receptors. PMID- 10385704 TI - Human and rat liver UDP-glucuronosyltransferases are targets of ketoprofen acylglucuronide. AB - Acylglucuronides formed from carboxylic acids by UDP-glucuronosyltransferases (UGTs) are electrophilic metabolites able to covalently bind proteins. In this study, we demonstrate the reactivity of the acylglucuronide from the nonsteroidal anti-inflammatory drug, ketoprofen, toward human and rat liver UGTs. Ketoprofen acylglucuronide irreversibly inhibited the glucuronidation of 1-naphthol and 2 naphthol catalyzed by human liver microsomes or by the recombinant rat liver isoform, UGT2B1, which is the main isoform involved in the glucuronidation of the drug. A decrease of about 35% in the glucuronidation of 2-naphthol was observed when ketoprofen acylglucuronide was produced in situ in cultured V79 cells expressing UGT2B1. Inhibition was always associated with the formation of microsomal protein-ketoprofen adducts. The presence of these covalent adducts within the endoplasmic reticulum of cells expressing UGT2B1 was demonstrated following addition of ketoprofen to culture medium by immunofluorescence microscopy with antiketoprofen antibodies. Immunoblots of liver microsomes incubated with ketoprofen acylglucuronide and probed with antiketoprofen antibodies revealed the presence of several protein adducts; among those was a major immunoreactive protein at 56 kDa, in the range of the apparent molecular mass of UGTs. The adduct formation partially prevented the photoincorporation of the UDP-glucuronic acid (UDP-GlcUA) analog, [beta-32P]5N3UDP-GlcUA, on the UGTs, suggesting that ketoprofen glucuronide covalently reacted with the UDP-GlcUA binding domain. Finally, UGT purification from rat liver microsomes incubated with ketoprofen glucuronide led to the isolation of UGT adducts recognized by both anti-UGT and antiketoprofen antibodies, providing strong evidence that UGTs are targets of this metabolite. PMID- 10385705 TI - Multiple amylin receptors arise from receptor activity-modifying protein interaction with the calcitonin receptor gene product. AB - Receptor activity-modifying proteins (RAMPs) are single-transmembrane proteins that transport the calcitonin receptor-like receptor (CRLR) to the cell surface. RAMP 1-transported CRLR is a calcitonin gene-related peptide (CGRP) receptor. RAMP 2- or RAMP 3-transported CRLR is an adrenomedullin receptor. The role of RAMPs beyond their interaction with CRLR, a class II G protein-coupled receptor, is unclear. In this study, we have examined the role of RAMPs in generating amylin receptor phenotypes from the calcitonin (CT) receptor gene product. Cotransfection of RAMP 1 or RAMP 3 with the human CT receptor lacking the 16 amino acid insert in intracellular domain 1 (hCTRI1-) into COS-7 cells induced specific 125I-labeled rat amylin binding. RAMP 2 or vector cotransfection did not cause significant increases in specific amylin binding. Competition-binding characterization of the RAMP-induced amylin receptors revealed two distinct phenotypes. The RAMP 1-derived amylin receptor demonstrated the highest affinity for salmon CT (IC50, 3.01 +/- 1.44 x 10(-10) M), a high to moderate affinity for rat amylin (IC50, 7.86 +/- 4.49 x 10(-9) M) and human CGRPalpha (IC50, 2.09 +/- 1.63 x 10(-8) M), and a low affinity for human CT (IC50, 4.47 +/- 0.78 x 10(-7) M). In contrast, whereas affinities for amylin and the CTs were similar for the RAMP 3-derived receptor, the efficacy of human CGRPalpha was markedly reduced (IC50, 1.12 +/- 0.45 x 10(-7) M; P <.05 versus RAMP 1). Functional cyclic AMP responses in COS-7 cells cotransfected with individual RAMPs and hCTRI1- were reflective of the phenotypes seen in competition for amylin binding. Confocal microscopic localization of c-myc-tagged RAMP 1 indicated that, when transfected alone, RAMP 1 almost exclusively was located intracellularly. Cotransfection with calcitonin receptor (CTR)I1- induced cell surface expression of RAMP 1. The results of experiments cross-linking 125I-labeled amylin to RAMP 1/hCTR transfected cells with bis succidimidyl suberate were suggestive of a cell surface association of RAMP 1 and the receptors. Our data suggest that in the CT family of receptors, and potentially in other class II G protein-coupled receptors, the cellular phenotype is likely to be dynamic in regard to the level and combination of both the receptor and the RAMP proteins. PMID- 10385706 TI - Levels of retinoic acid and retinaldehyde dehydrogenase expression in eyes of the Mitf-vit mouse model of retinal degeneration. AB - PURPOSE: Several reports have characterized the retinal degeneration observed in the Mitf(vit) mutant mouse. Despite these reports, the factor(s) that may cause or modulate the degeneration still are not well defined; however, it is known that the photoreceptors of Mitf(vit) mice die through an apoptotic mechanism. We reported previously that retinoid metabolism in the RPE of Mitf(vit)++ mice is perturbed. Retinoids regulate genes via the RAR and RXR nuclear receptor pathway that are involved in numerous cellular responses including apoptosis. It is possible that retinoic acid (RA) modulates the retinal degeneration observed in the Mitf(vit) mice. The purpose of this study was to evaluate the levels of RA in whole eyes, as well as its distribution between neural retina and RPE, of the Mitf(vit) mutant mouse model. An additional purpose was to examine the expression of the RA generating enzyme, retinaldehyde dehydrogenase (AHD2), in the eyes of mutant and control mice. METHODS: The distribution of AHD2 in eyes of pre- and postnatal Mitf(vit) and C57BL/6 wild-type mice was determined immunohistochemically. Quantitative and qualitative analyses of RA were performed using reversed-phase high performance liquid chromatography (HPLC). RESULTS: The distribution of AHD2 in ocular tissues was similar between pre- and postnatal Mitf(vit) and C57BL/6 control mice. At postnatal week 10, however, a marked increase in AHD2 immunoreactivity was noted in the central dorsal neural retina of Mitf(vit) mice. No differences in the level of total RA in whole eyes were noted between Mitf(vit) and control mice at early postnatal ages. By 10 weeks of age there was a significant elevation of RA that was localized to the neural retina. CONCLUSIONS: In this study, we show a high level of AHD2 and RA in the neural retina of Mitf(vit) mice relative to control mice. It is possible that this elevation of RAs contributes to the retinal degeneration observed in Mitf(vit) mice either by inducing apoptosis or by enhancing the effect of some other factor(s) involved in the apoptotic pathway. PMID- 10385707 TI - A systems approach in hepatology. AB - Systems analysis has been applied to hepatology with the aim of providing a reasonable organisation of domain knowledge and supporting the improvement of clinical performance. To this extent liver structures and functions have been classified and defined, clinical parameters have been carefully selected and suitably associated to describe individual liver functions, and methodological criteria for clinical evaluation have been assessed. Three major outcomes of such an approach, respectively concerning the development of a shareable clinical database, the application of a suitable methodology for clinical reasoning, and the computer-based support to medical decision-making, have been discussed. PMID- 10385708 TI - Non-invasive diagnosis and management of chronic liver diseases. AB - Advances in computer-aided diagnosis, imaging techniques, DNA mutation analysis, virology, immunology and biochemistry have improved our understanding of chronic liver diseases and the possibilities for non-invasive diagnosis. Various medical therapies for chronic liver diseases and their complications have been developed recently, and their monitoring has also improved. This review focuses on these recent advances in non-invasive diagnosis and management of chronic liver diseases. PMID- 10385709 TI - Scoring procedures in clinical hepatology. AB - A major problem in the assessment of liver function is represented by the quantification of the different aspects on which it relies (biosynthesis, drug metabolism, bile secretion, etc.) and of the clinical severity, with important prognostic implications. Another field that can be supported by quantification procedures is the histological evaluation of chronic hepatitis (necro inflammatory activity and fibrosis). Finally, scoring systems can be usefully applied in clinical practice as a tool which supports medical decisions in very difficult diagnostic processes. In all the above considered fields, the scoring procedures have the important advantage to allow the standardisation of clinical procedures as well as to facilitate the statistical manipulation of data in controlled clinical trials. This paper reviews numerical scoring systems utilised in hepatology and their clinical applications. PMID- 10385710 TI - Liver transplantation: who to refer and when. AB - Liver transplantation is a surgical procedure offered to individuals with irreversible, near fatal liver disease. The timing of both transplantation listing and surgical engraftment are critical factors in the success of this endeavour. To accomplish each and maintain surgical survival rates without prematurely transplanting individuals to achieve excellent outcome statistics is an art that requires knowledge about the procedure and the natural history of the specific liver disease in question. Herein are the views of the transplant team at Loyola University of Chicago as to how this can be accomplished within the framework of the American experience, and the rules and regulations governing donor organ procurement and allocation in the United States. PMID- 10385711 TI - Thyrotropin receptor: a role for thyroid tumourigenesis? AB - Human thyroid tumours represent an example of the interplay of genetic and non genetic carcinogenesis. Recently, genetic abnormalities in the elements of the Thyrotropin receptor (TSH-R) dependent cAMP regulatory cascade have been found to be involved both in benign and malignant thyroid tumours. The presence of activating mutations has been demonstrated in the TSH-R gene as well as in the Gs alpha protein gene in thyroid toxic adenoma resulting in the constitutive activation of the cAMP pathway and it has been hypothesised that these genetic alterations may play a causative role in the disease. However, recent observations suggest more caution in accepting such a hypothesis. The presence of activating TSH-R mutations has also been demonstrated in differentiated thyroid carcinomas. At present, the percentage of such a modification is low, unless referred to selected series of tumours. Activating mutations of the TSH-R gene have been detected in a group of differentiated carcinomas with high basal adenylyl cyclase activity, and in a few cases of hyperfunctioning thyroid carcinoma. However, the role of the TSH-R-related cAMP pathway alterations in thyroid transformation remains to be elucidated. In this review, the role of TSH R gene alterations in benign and malignant thyroid neoplasia is examined. PMID- 10385712 TI - Relationship between ideology and science. AB - After defining the characteristics of ideological knowledge and knowledge based on research, the experimental work on illusory correlations, serial effects, difficulties in "grasping counter-examples" and prejudiced pseudo-knowledge is reported. This proves how ideology can develop from the very functioning of the cognitive processes (perception, thought) when it is not kept under critical scrutiny. The difference between ideological and scientific thought reflects the different social conditions behind the production of the two types of knowledge. The production of scientific knowledge is regulated by specific rules such as the logic of the experimental method and empirical references, and is animated by a depressive attitude ("I am responsible for matters within the confines of rules set by the research community") while the propositions of ideology are anti empirical, shy away from counter-examples, are confusional, and are underpinned by an attitude that is potentially maniacal and omnipotent. The ideological factors that can have an effect on professional research are listed and it is shown how the results of this, once controlled, lose all ties to the ideology they may have been inspired by, to the extent that they constitute another sphere that is completely autonomous and independent. PMID- 10385713 TI - Randomized trial of ligation versus combined ligation and sclerotherapy for bleeding esophageal varices. AB - BACKGROUND: Endoscopic band ligation combined with sclerotherapy has been postulated to be superior to ligation alone for the treatment of esophageal variceal bleeding. METHODS: A randomized trial of ligation versus combined ligation and sclerotherapy was designed to determine whether combined therapy results in faster eradication of varices compared to ligation alone. Sixty patients were randomized to undergo band ligation or ligation combined with injection of 1 to 2 mL of polidocanol (1%) into each variceal column immediately proximal to the previously placed bands. Therapy was repeated at 1- or 2-week intervals until variceal eradication was achieved. Follow-up endoscopy was performed at 3 months and then at 6-month intervals. RESULTS: The demographic and clinical characteristics of the 31 patients who underwent ligation were similar to those of the 29 who received combined treatment. Sixty percent of the patients had cirrhosis due to viral hepatitis. No significant differences were found between the combined and ligation alone groups in arresting active bleeding [9 of 9 (100%) vs. 6 of 7 (86%)], units of blood transfusion (3 +/- 0.8 vs. 2 +/- 0.6), number of sessions required to eradicate varices (3.8 +/- 0.5 vs. 3.6 +/- 0.4), treatment failure [2 (17%) vs. 4 (14%)], esophageal varix recurrence [6 (21.%) vs. 2 (6%)], gastric varices formation [4 (14%) vs. 1 (3%)], stricture [1 (3%) vs. 0 (0%)], recurrent bleeding [5 (17%) vs. 7 (23%)], other complications [10 (34%) vs. 9 (29%)], or death [3 (10%) vs. 7 (23%)] during a follow-up period of up to 36 months. CONCLUSIONS: Combined ligation and sclerotherapy does not reduce the number of endoscopic treatment sessions required for variceal eradication and offers no benefit over ligation alone. Because of the lack of benefit, the added procedure time, and the cost, we do not advocate combination therapy, and ligation alone remains the best endoscopic treatment. PMID- 10385715 TI - A randomized prospective study comparing rigid to balloon dilators for benign esophageal strictures and rings. AB - BACKGROUND: The optimum choice of dilator (rigid vs. balloon) for benign esophageal strictures has not been well studied. The aim of this study was to compare the immediate relief of dysphagia and the incidence of repeat dilatation within the first year with the use of either a rigid (Savary) dilator or balloon dilator for benign lower esophageal strictures. METHODS: Patients with dysphagia found to have benign esophageal strictures during endoscopy were randomized to undergo dilation with a rigid (Savary) or a balloon dilator (Microvasive or Bard). The 1-year incidence of repeat dilatation was estimated by the Kaplan Meier method. RESULTS: A total of 251 subjects were stratified at entry according to the type of stricture (peptic vs. Schatzki ring) and severity of stricture (mild vs. moderate/severe) and then randomized to either a Savary (n = 88), Microvasive (n = 81), or Bard (n = 82) dilator. There were no significant differences between the rigid dilator or the two balloons with regard to immediate relief of dysphagia or the need for repeat dilatation at one year. Patients with moderate/severe strictures required repeat dilatation at one year twice as often as those with mild strictures. There were no significant complications reported in these patients. CONCLUSIONS: Both rigid and balloon dilators are equally effective and safe in the treatment of benign lower esophageal strictures caused by acid reflux and Schatzki rings. PMID- 10385714 TI - Sclerotherapy plus ligation versus ligation for the treatment of esophageal varices: a prospective randomized study. AB - BACKGROUND: We devised a new combined method of endoscopic variceal ligation and injection sclerotherapy, namely, endoscopic scleroligation, for the treatment of esophageal varices. The aim of this prospective randomized trial was to compare endoscopic scleroligation with endoscopic variceal ligation alone with regard to efficacy, complications, variceal recurrence, and survival. METHODS: Fifty-one patients with cirrhosis and esophageal varices were randomly assigned to be treated by endoscopic scleroligation (n = 25) or endoscopic variceal ligation (n = 26). In the initial session in the endoscopic scleroligation group, endoscopic injection sclerotherapy was performed with injection of 5% ethanolamine oleate around the lower esophagus to obliterate the feeding veins. This was followed by endoscopic variceal ligation from the injection site to the most orad varix. In subsequent sessions, endoscopic injection sclerotherapy was performed with 1% polidocanol. In the endoscopic variceal ligation group, that procedure was performed in all treatment sessions. RESULTS: Both methods were equally effective in achieving complete eradication of esophageal varices. Among the cases in which complete eradication was achieved, the 1- and 3-year cumulative recurrence rates in the endoscopic scleroligation group (9.5%, 22.1%) were significantly lower than those in the endoscopic variceal ligation group (61.9%, 72.2%) (p < 0.01). The survival rates and incidences of treatment-related complications have been similar among patients treated by both methods. CONCLUSIONS: Endoscopic scleroligation is superior to endoscopic variceal ligation in preventing variceal recurrence. PMID- 10385716 TI - The efficacy and safety of argon plasma coagulation therapy in Barrett's esophagus. AB - BACKGROUND: Thermoablation is being used to eliminate the metaplastic epithelium of Barrett's esophagus and allow its reversal into squamous epithelium in an acid controlled environment. This study assessed the efficacy and safety of a new thermoablation technique, argon plasma coagulation. METHODS: Patients with circumferential Barrett's esophagus 2 to 5 cm long were enrolled. Acid suppression was accomplished with lansoprazole. One-half the circumference of Barrett's mucosa was treated with argon plasma coagulation, and the other half served as an internal control. After macroscopic squamous re-epithelialization occurred, biopsy specimens were obtained from both areas systematically. RESULTS: Nine patients, all men with a mean age of 51.1 years, completed the study. During 24-hour esophageal pH monitoring a pH less than 4 occurred on average 2.8% of the time with a mean dose of lansoprazole of 70 mg/day. Squamous re-epithelialization developed in treated areas in all 9 patients. Biopsy showed that 7 of 9 patients (77.8%) had squamous re-epithelialization without intestinal metaplasia. Biopsy showed that 2 of 9 patients (22.2%) had squamous re-epithelialization with evidence of underlying intestinal metaplasia. There were no serious complications. CONCLUSIONS: Argon plasma coagulation in an acid-controlled environment was both efficacious and safe in the treatment of Barrett's esophagus. However, the reappearance of squamous epithelium after therapy did not exclude the presence of underlying intestinal metaplasia. PMID- 10385717 TI - Variable pathologic interpretation of columnar lined esophagus by general pathologists in community practice. AB - BACKGROUND: Pathologic interpretation of biopsy specimens of columnar lined esophagus guides subsequent endoscopic surveillance and/or surgical intervention. The aim of this study was to evaluate pathologic interpretation of columnar lined esophagus by general pathologists in community practice. METHODS: Five histologic slides representing different types of columnar lined esophagus were submitted for review by 20 randomly selected general pathologists in community practice. There were three cases with intestinal metaplasia (one with no dysplasia, one with low-grade dysplasia, and one with high-grade dysplasia) and two cases of gastric metaplasia (one fundic-type and one cardia-type). RESULTS: High-grade dysplasia was identified as such by 30% of pathologists and was called invasive adenocarcinoma by 20%, low-grade dysplasia by 30%, and moderate dysplasia by the remaining 20%. Low-grade dysplasia was identified as such by 35% of pathologists and was called high-grade dysplasia by 20%, moderate dysplasia by 20%, and no dysplasia by 25%. Specialized columnar epithelium with no dysplasia was identified as such by 35%, called low-grade dysplasia by 35%, moderate dysplasia by 15%, indeterminate for dysplasia by 10%, and invasive adenocarcinoma by 5%. Gastric metaplasia without specialized columnar epithelium was identified as Barrett's esophagus in 38% of cases. CONCLUSIONS: Pathologic interpretation of columnar lined esophagus by community pathologists may be subject to marked interobserver variation. The term Barrett's esophagus is often used to describe columnar lined esophagus without goblet cells. Because this finding is not clearly associated with an increased risk of cancer, these data support recent suggestions that the term Barrett's esophagus be abandoned. Interpretations of both high-grade and low-grade dysplasia should be considered for review by experts in esophageal pathology. PMID- 10385719 TI - Role of EUS in the management of pancreatic and ampullary carcinoma: a prospective study assessing resectability and prognosis. AB - BACKGROUND: Endoscopic ultrasonography (EUS) is highly accurate for the staging of tumors, but its role in the management of periampullary carcinoma is still being defined. METHODS: Seventy-nine patients with pancreatic (n = 73) or ampullary (n = 6) carcinoma underwent prospective evaluation by means of assessment of resectability and survival according to the following three-step staging algorithm: (1) ultrasonography and computed tomography; (2) if tumor appears resectable, EUS; (3) if criteria of resectability are found at EUS, laparotomy for curative resection. RESULTS: The first step of the algorithm helped predict unresectability of tumors and need for palliative treatment for 36 patients. Among the other 43 patients EUS revealed signs of unresectability in 20 additional patients who then underwent palliative surgical or medical treatment (median survival time 7 to 8 months). Twenty-three carcinomas were considered resectable according to EUS findings: Palliative surgery was performed in 9 cases (survival time 6 months), and 14 tumors could be resected in a curative way with a median survival period of 15 (pancreatic) to 16 months (ampullary). In evaluation of resectability, EUS had a 50% sensitivity (positive examination), 100% specificity, 100% positive predictive value, 61% negative predictive value, and 72% accuracy. CONCLUSIONS: EUS is accurate for evaluating resectability of ampullary and pancreatic cancer. EUS staging can prevent unnecessary surgery, and the findings correlate well with prognosis. The management of ampullary and pancreatic cancer could be improved with EUS. PMID- 10385718 TI - EUS compared with CT, magnetic resonance imaging, and angiography and the influence of biliary stenting on staging accuracy of ampullary neoplasms. AB - BACKGROUND: Computerized tomography (CT), magnetic resonance imaging (MRI), and transabdominal ultrasound frequently fail to detect ampullary lesions. Endoscopic ultrasound (EUS) is a sensitive modality for detecting and staging ampullary tumors. Accurate staging may be affected by biliary stenting, which is frequently performed in these patients with obstructive jaundice. The present study assessed the accuracy of ampullary tumor staging with multiple imaging modalities in patients with and those without endobiliary stents. METHODS: Fifty consecutive patients with ampullary neoplasms from two endosonography centers were preoperatively staged by EUS plus CT (37 patients), MRI (13 patients), or angiography (10 patients) over a 3(1/2) year period. Twenty-five of the 50 patients had a transpapillary endobiliary stent present at the time of endosonographic examination. Accuracy of EUS, CT, MRI, and angiography was assessed with the TNM classification system and compared with surgical-pathologic staging. The influence of an endobiliary stent present at the time of EUS on staging accuracy of EUS was also evaluated. RESULTS: EUS was more accurate than CT and MRI in the overall assessment of the T stage of ampullary neoplasms (EUS 78%, CT 24%, MRI 46%). No significant difference in N stage accuracy was noted between the three imaging modalities (EUS 68%, CT 59%, MRI 77%). EUS T stage accuracy was reduced from 84% to 72% in the presence of a transpapillary endobiliary stent. This was most prominent in the understaging of T2/T3 carcinomas. CONCLUSIONS: EUS is superior to CT and MRI in assessing T stage but not N stage of ampullary lesions. The presence of an endobiliary stent at EUS may result in underestimating the need for a Whipple resection because of tumor understaging. PMID- 10385720 TI - Catheter probe assisted endoluminal US in inflammatory bowel disease. AB - BACKGROUND: Use of an echocolonoscope to examine patients with inflammatory bowel disease is technically difficult. Catheter probe assisted endoluminal ultrasonography (US) may be a feasible alternative. METHODS: Determination of demographic information and clinical disease activity was followed by colonoscopy with biopsy. Catheter probe assisted endoluminal US was performed with measurements of thickness of the intestinal wall and evaluation of the structure of the sonographic layers. RESULTS: Twenty-eight patients, 7 with ulcerative colitis, 11 with Crohn's disease, and 10 healthy control subjects participated in a prospective study. Mean colonic wall thickness was 2.2 +/- 0.1 mm (controls) compared with 4. 1 +/- 0.4 mm (ulcerative colitis) (p < 0.001) and 4.4 +/- 0.4 mm (Crohn's disease) (p < 0.001). Among patients with ulcerative colitis, colonic wall thickness correlated with severity of colonoscopic changes (r = 0.84, p = 0.02). Among patients with Crohn's disease, loss of endosonographic layer structure correlated with disease activity score (r = 0.8, p = 0.003), and colonic wall thickness correlated with the severity of histologic changes (r = 0. 62, p = 0.04). CONCLUSIONS: Catheter probe assisted endoluminal US is technically feasible in the care of patients with inflammatory bowel disease. Endosonographic measurements of colonic wall thickness and layer structure provide clinically significant information. PMID- 10385722 TI - Increased rate of complete EUS staging of patients with esophageal cancer using the nonoptical, wire-guided echoendoscope. AB - BACKGROUND: Incomplete endoscopic ultrasound (EUS) staging procedures in patients with esophageal cancer due to obstructing malignant strictures are prone to underestimate T stage and cannot detect celiac adenopathy. EUS staging in the setting of stenotic malignancies using the large caliber echoendoscope has been complicated by esophageal perforation. We report on the clinical utility of a newly developed, wire-guided echoendoscope for the complete staging of patients with esophageal cancer. METHODS: Pretreatment EUS examinations performed for esophageal cancer staging were evaluated and the ability to traverse the esophagus and examine the celiac axis were documented. Outcomes before and after the availability of the wire-guided echoendoscope were compared. RESULTS: One hundred thirty consecutive examinations were evaluated 100 before and 30 after the introduction of the wire-guided echoendoscope. Complete staging was accomplished in 60 of 100 (60%) cases before and 27 of 30 (90%) after its introduction (p = 0.002). The wire-guided echoendoscope was used in 14 of the 30 cases. Despite a trend toward fewer stage T4 tumors, metastatic disease was documented significantly more frequently after the introduction of the esophagoprobe (34% vs. 11%, p = 0.002). There were no complications. CONCLUSIONS: The introduction of the wire-guided echoendoscope markedly reduced the occurrence of incomplete esophageal cancer staging and improved the detection of metastatic disease. PMID- 10385721 TI - Use of color Doppler EUS in assessing azygos blood flow for patients with portal hypertension. AB - BACKGROUND: Azygos blood flow is an index of blood flow through gastroesophageal collateral vessels and varices in portal hypertension. Conventional measurement of azygos blood flow involves catheterization of the azygos vein. We studied the feasibility of assessing azygos blood flow with color Doppler endosonography and of monitoring the effects of vasoactive agents on azygos blood flow. METHODS: Patients with portal hypertension were examined by means of linear array color Doppler endoscopic ultrasonography (EUS). Patients who had taken propranolol or nitrates in the 4 weeks before the day of measurement of azygos blood flow were excluded. After identification of the azygos vein and recording of baseline readings of mean arterial blood pressure, pulse rate, and azygos blood flow, patients were selected in a random manner to receive a bolus injection of 2 mg terlipressin, 250 microg somatostatin, or saline solution (control). Azygos blood flow was measured 1, 5, and 10 minutes after injection (AzBF-1, AzBF-5, AzBF-10). RESULTS: Six patients were recruited in each treatment group. Basal azygos blood flow showed a positive association with the Child-Pugh grade of cirrhosis (p < 0.005). After bolus injection of terlipressin and somatostatin, there was a marked decrease in AzBF-1 (24% and 37%), AzBF-5 (42% and 19%), and AzBF-10 (40% both) compared with baseline. The control group showed no significant change in azygos blood flow. CONCLUSIONS: Color Doppler EUS is useful in assessing azygos blood flow in portal hypertension and in monitoring the effects of vasoactive agents. PMID- 10385723 TI - Helicobacter pylori status and endoscopy follow-up of patients having a history of perforated duodenal ulcer. AB - BACKGROUND: The aim of this study was to determine whether the recurrence of symptoms or ulcer disease in patients with a history of perforated duodenal ulcer is related to Helicobacter pylori infection. METHODS: One hundred sixty-three consecutive patients with history of perforated duodenal ulcer unrelated to nonsteroidal anti-inflammatory drugs underwent upper endoscopy. Any recurrent symptoms or complications were documented. Regardless of the endoscopic findings, three antral biopsy specimens were taken for histologic examination and a rapid urease test. RESULTS: There was a preponderance of men (male/female = 5.3:1). The mean age was 55.9 years. Sixty-seven (41.1%) patients gave a history of recurrent epigastric pain, seven of whom also had a history of bleeding ulcer. Upper endoscopy was performed at a mean of 74.5 +/- 7.1 months after operation. Positive endoscopic findings were noted in 68 (41.7%) patients; H. pylori was found in the biopsy specimens from 77 (47.2%) patients. Recurrent duodenal ulcer was found in 29 (17.8%) patients and was significantly related to male gender, recurrent epigastric pain, bleeding ulcer, longer interval from previous operation, and positive H. pylori status. Positive H. pylori status and male gender were independent factors associated with recurrent duodenal ulcer. CONCLUSIONS: Recurrent ulcer disease in patients with a history of perforated duodenal ulcer is related to H. pylori infection. PMID- 10385724 TI - Outpatient therapeutic ERCP with endobiliary stent placement for malignant common bile duct obstruction. AB - BACKGROUND: Since 1996 patients in stable condition who need therapeutic endoscopic retrograde cholangiopancreatography (ERCP) at our institution have been treated as outpatients whenever possible. We reviewed our institution's experience and compared outpatient versus inpatient therapeutic ERCP for endobiliary stent placement in the care of patients with malignant common bile duct obstruction. METHODS: A retrospective review of all therapeutic ERCPs for the palliation of malignant common bile duct obstruction with endobiliary stents was performed from March 1, 1996, through December 1, 1997. RESULTS: One hundred nine therapeutic ERCPs were performed on 84 patients to place a polyethylene endobiliary stent for malignant common bile duct obstruction. Forty-three procedures were performed on 31 outpatients, 66 on 53 inpatients. There was no significant difference between outpatient and inpatient groups with regard to age, gender, procedure success rate, complication rate, need for endoscopic sphincterotomy, or whether the procedure was for initial stent placement or stent exchange. Inpatients had no procedure-related complications; outpatients had two. There was no procedure-related mortality in either group. CONCLUSION: Therapeutic ERCP for palliation of malignant common bile duct obstruction can be safely and successfully performed on an outpatient basis for selected patients. This should result in better quality of life for these patients with advanced cancer and substantial cost savings. PMID- 10385725 TI - The role of choledochoscopy in the diagnosis and management of biliary tract diseases. AB - BACKGROUND: The diagnosis and management of biliary tract disorders in certain cases may be incomplete without direct visualization of the bile ducts. METHODS: We report our experience of 61 choledochoscopies (33 women, 27 men, mean age 44.6 years). Twenty patients had previously undergone orthotopic liver transplantation. All except two choledochoscopies were performed via the transpapillary route. Indications included suspected large bile duct stones in 18 patients, anastomotic strictures in 16, abnormal cholangiograms in 5, elevated liver function tests in 7, suspected cholangiocarcinoma in 4, occluded biliary metallic stent in 4, hemobilia in 4, primary sclerosing cholangitis in 2 and ischemic bile duct injury in 1 patient. RESULTS: Choledochoscopy confirmed the anticipated diagnosis in 36 of 61 (59%) patients. Importantly, it provided additional unsuspected diagnostic information in 18 of the 61 (29.5%) patients. In addition, for patients in whom standard cholangiography was deemed abnormal, choledochoscopy demonstrated normal results in 7 (11.4%) patients. Fifty-two choledochoscopies were performed with therapeutic intentions, and the procedure was helpful in providing targeted treatment in 27 (44.2%) patients. CONCLUSIONS: Choledochoscopy is a safe and useful endoscopic modality that can provide specific diagnoses and direct treatment in various biliary tract diseases. The additional information provided by choledochoscopy may change overall patient management and outcome. PMID- 10385726 TI - Does endoscopic papillary balloon dilation affect gallbladder motility? AB - BACKGROUND: Endoscopic papillary balloon dilation for treatment of bile duct stones is likely to preserve papillary function. However, endoscopic papillary balloon dilation may affect gallbladder motility. We investigated the effects of endoscopic papillary balloon dilation on gallbladder motility. METHODS: Ten patients with an intact gallbladder (six with and four without gallbladder stones) who underwent endoscopic papillary balloon dilation for choledocholithiasis were studied. Gallbladder motility was examined before and 7 days and 1 month after endoscopic papillary balloon dilation. Gallbladder volume, while fasting and after dried egg yolk ingestion, was determined by ultrasonography. RESULTS: Before endoscopic papillary balloon dilation, particularly in patients with gallbladder stones, the gallbladder showed significantly larger fasting volume and lower yolk-stimulated maximum contraction compared with control subjects. Seven days after endoscopic papillary balloon dilation, fasting volume was decreased and maximum contraction was increased, regardless of the presence of gallbladder stones, with significant differences from the values before endoscopic papillary balloon dilation. One month after endoscopic papillary balloon dilation, these changes were reduced and gallbladder function did not differ significantly from baseline. CONCLUSIONS: After endoscopic papillary balloon dilation, gallbladder motility improves transiently at 7 days but returns to baseline at 1 month. In terms of gallbladder motility, endoscopic papillary balloon dilation does not seem to increase the subsequent risk of acute cholecystitis. PMID- 10385727 TI - Iodine absorption in patients undergoing ERCP compared with coronary angiography. AB - BACKGROUND: Systemic absorption of iodinated contrast material occurs during endoscopic retrograde cholangiopancreatography (ERCP), the clinical significance of which has not yet been determined. METHODS: Urinary iodine excretion was measured before and after coronary angiography (n = 20) and ERCP (n = 12). Thyroid hormone levels were determined before iodine load and after 6 and 24 weeks. RESULTS: Before coronary angiography, iodine excretion was 101 +/- 38.3 micromol/mol creatinine and increased to 865. 10(5) +/- 721. 10(5) micromol/mol on the next day (p 60 years of age, patients with diabetes, patients with congestive heart failure, and patients taking angiotensin converting enzyme inhibitors. Mean follow-up period was 16 months. Ten patients died during follow-up: 9 of cardiac causes and 1 of stroke. Nine of these 10 patients had heart rate variability <10 beats/min (P =.004). The sensitivity and specificity of this test for cardiovascular mortality is 90.0% and 68.0%, respectively. The negative predictive value is 99.2% and the relative risk is 16.6. Heart rate variability <10 beats/min remained a significant predictor of death after adjusting for clinical, demographic, and left ventricular function with an odds ratio of 1.38 (95% confidence interval, 1.13-1.63). CONCLUSIONS: This simple, brief bedside deep breathing test of heart rate variability in patients after myocardial infarction appears to be a good predictor for all-cause mortality and sudden death. It may be used as a clinical test for risk stratification after myocardial infarction. PMID- 10385762 TI - Prehospital testing for troponin T in patients with suspected acute myocardial infarction. AB - BACKGROUND: Cardiac troponin T (TnT) is a highly sensitive and specific marker for myocardial damage and can be detected early after myocardial injury. Our hypothesis was to use TnT as an objective marker to verify acute myocardial infarction before hospital admission. METHODS AND RESULTS: We evaluated the sensitivity of a rapid qualitative assay for serum TnT for the detection of acute myocardial infarction in the ambulance and assessed the predictive value of a positive prehospital TnT test for death and myocardial infarction during 6-months of follow-up. The study, conducted in an urban area, included 158 consecutive patients with suspected acute myocardial infarction (93 men aged 69 +/- 13 years). A myocardial infarction was confirmed in 40 and excluded in 118 patients. The prehospital TnT test was positive in 11 patients, of whom 7 had acute myocardial infarction. Fifty-three patients had a positive test result at hospital admission, with evidence of myocardial infarction in 39 of them. The sensitivity to acute myocardial infarction was 18% for the prehospital and 98% for the in-hospital test with 78% and 88% specificity, respectively. During follow-up, patients with a positive prehospital TnT test result had cardiac events more often (9 of 11) than patients with a negative result (26 of 147; P <.0001). CONCLUSIONS: In areas with short transport times to the patient the rapid TnT test performed at the point of care identified only a minority of the patients with acute myocardial infarction. A positive prehospital TnT test result seems to be an objective marker for a worse outcome in patients presenting with suspected acute myocardial infarction. PMID- 10385761 TI - Hemodynamic effects of double bolus reteplase versus alteplase infusion in massive pulmonary embolism. AB - BACKGROUND: Thrombolytic agents are given in massive pulmonary embolism to dissolve or reduce the clot and normalize hemodynamics. Comparative clinical studies have shown that administration of a 2-hour infusion of alteplase is more effective than urokinase over a 12-hour period. Reteplase is a new generation thrombolytic with a longer half-life that can be administered more conveniently as a double bolus. We compared efficacy and safety of reteplase with the approved regimen of alteplase in massive pulmonary embolism. METHODS: Thirty-six patients were enrolled and randomly assigned: 23 received reteplase and 13 received alteplase along with intravenous heparin. Reteplase was administered as 2 intravenous bolus injections of 10 U 30 minutes apart, and alteplase was administered as an intravenous infusion of a total dose of 100 mg over a 2-hour period, including an initial 10-mg bolus. Diagnosis of pulmonary embolism was confirmed by selective pulmonary angiography. Hemodynamic monitoring was conducted during the first 24 hours after administration. The primary end point was change in total pulmonary resistance. Secondary variables were pulmonary pressure, cardiac index, clinical parameters, and adverse events. RESULTS: The primary parameter of total pulmonary resistance showed a significant decrease after just 0.5 hours in the reteplase group and after 2 hours in the alteplase group, with a further decrease persisting for up to 24 hours in both treatment groups. A similar pattern was seen in other directly measured hemodynamic parameters, especially mean pulmonary artery pressure and cardiac index; there was no significant difference between reteplase and alteplase. There was also no apparent difference between the treatment groups with respect to safety, and no stroke or intracranial hemorrhage occurred. The rate of bleedings and the incidence of nonhemorrhagic adverse events were as expected for patients with pulmonary embolism treated with a thrombolytic agent. CONCLUSIONS: Reteplase is suitable for treatment of massive pulmonary embolism with a standard double bolus 10 + 10 U. Efficacy of reteplase appeared to be at least as good at decreasing pulmonary vascular resistance as that of the approved alteplase regimen of 100 mg infusion over a 2-hour period. PMID- 10385763 TI - Effects of platelet glycoprotein IIb/IIIa inhibition with abciximab on thrombin generation and activity during percutaneous coronary intervention. AB - BACKGROUND: Antagonists of the platelet glycoprotein IIb/IIIa decrease acute ischemic complications after percutaneous coronary interventions (PCI). Abciximab (c7E3 Fab, ReoPro) has been reported to decrease thrombin generation in vitro. We investigated in vivo the effect of abciximab therapy on thrombin generation, thrombin activity, and the activated clotting time (ACT) during PCI. METHODS: We studied 32 consecutive patients who underwent PCI for unstable coronary syndromes. Group I (n = 11) was treated with heparin plus aspirin, and group II (n = 21) was treated with heparin plus aspirin plus standard-dose abciximab, administered 5 minutes after the initial heparin bolus. Patients received a standardized heparin bolus at time 0, and arterial blood specimens for prothrombin fragment F1.2, fibrinopeptide A (FPA), and ACT were obtained from the guiding catheter at 5 minutes, 10 minutes (ACT only), 20 minutes, and at the end of the PCI. Standard-dose abciximab was administered in group II only. Each patient served as his or her own control, and the changes against the baseline were compared between the 2 groups. RESULTS: There were no significant differences between the 2 groups regarding baseline characteristics, hematocrit, and platelet count. Group I patients had higher ACT and lower F1.2 and FPA compared with group II at baseline. Subsequent measurements demonstrated a gradual decrease in FPA and F1.2 in group II; the end of procedure versus baseline changes that occurred in F1.2 were significantly different compared with group I (decrease of 0.59 +/- 0.22 nmol/L in group II vs increase of 0.22 +/- 0.3 nmol/L in group I, P =.04), and a trend in the same direction was evident for FPA changes (decrease of 1.46 +/- 1.16 ng/mL in group II vs increase of 2.25 +/- 1.58 ng/mL in group I, P =.07). The ACT response to abciximab was variable, but a 6.3% increase (+20 sec) in ACT was documented 5 minutes after abciximab bolus in group II compared with the 3.4% decrease (-10 sec) observed in group I at the same time point (P =.1). CONCLUSION: Addition of abciximab to heparin plus aspirin during PCI was associated with a significant decrease in thrombin generation and a borderline decrease in thrombin activity. PMID- 10385764 TI - Direct coronary stenting without balloon predilation in acute coronary syndromes. AB - BACKGROUND: This prospective, observational study was designed to evaluate the feasibility and the safety of a new strategy for stenting in acute coronary syndromes: direct stent implantation without predilation of the culprit lesion. This strategy might reduce both the cost of the procedure and the rate of no reflow, a phenomenon that is more frequently observed during dilation of unstable plaques. METHODS AND RESULTS: Between September 1997 and March 1998, 122 carefully selected patients with unstable angina or acute myocardial infarction were included in this study. Highly calcified lesions and vessels with excessive proximal tortuosity were excluded. The procedure was successful in 96% of cases. In 5 cases the stent failed to pass through the stenosis and was successfully retrieved in the guiding catheter in 3 cases. In 2 cases the stent was lost in the peripheral circulation. Transient no-reflow occurred in only 3 cases and was rapidly reversed by rescue use of an intracoronary bolus injection of a glycoprotein IIb/IIIa receptor inhibitor in 2 cases. A patient treated by primary angioplasty with cardiogenic shock on admission died 48 hours after the initial procedure because of irreversible cardiac failure. One-month clinical follow-up was obtained by telephone for all patients; no major coronary events occurred during this period. CONCLUSIONS: Direct coronary stenting without balloon predilation can be safely performed in acute coronary syndrome-related lesions in selected patients. A randomized, controlled study is warranted to confirm the promising results of this pilot study, especially regarding the low rate of the no-reflow phenomenon. PMID- 10385765 TI - Prospective randomized trial of corticosteroids for the prevention of restenosis after intracoronary stent implantation. AB - BACKGROUND: Inflammatory responses have been implicated as one of the major contributors to neointimal hyperplasia after coronary stenting. The aim of this study was to evaluate the effect of pretreatment with single-dose intravenous methylprednisolone on preventing in-stent restenosis. METHODS: One hundred and forty consecutive patients for elective coronary stenting (focal, de novo lesion and reference diameter >/=3 mm) were randomly assigned to either a methylprednisolone or a placebo group. Either 1 g methylprednisolone or placebo was intravenously infused 6 to 12 hours before stenting with one of two types of stents. Follow-up angiography was performed at 6 months and clinical evaluation made at regular intervals. RESULTS: Baseline characteristics were similar between both groups. Stenting was successful in all patients, and in-hospital events did not occur in any patients. Follow-up angiography was performed in 127 patients (follow-up rate of 91.4%). The minimal lumen diameter increased from 0.86 +/- 0.50 mm before intervention to 3.34 +/- 0.42 mm after intervention (P =.02). At follow-up, minimal lumen diameter decreased to 2.14 +/- 0.78 mm (P <. 01). Angiographic restenosis rate was 17.5% in the steroid group and 18.8% in the placebo group (P =.85), with no differences between the 2 types of stent. Clinical follow-up was available in all patients (10.3 +/- 2.5 months) and clinical events during the follow-up period were similar in both groups. CONCLUSIONS: Single-dose pretreatment with intravenous methylprednisolone before coronary stenting had no effect on the change in minimal lumen diameter at a mean follow-up time. PMID- 10385766 TI - Administration of protamine after coronary stent deployment. AB - BACKGROUND: Prompt reversal of anticoagulation by protamine administration could be an important therapeutic option to treat serious procedural complications such as vessel rupture or major bleeding from the puncture site during coronary stent implantation. However, this therapeutic option is rarely used because of the possible risk of stent thrombosis. METHODS: We retrospectively analyzed the incidence of acute and subacute stent thrombosis and vascular complications in 90 patients who received protamine (protamine group) and 1763 patients who did not receive protamine (control group) after successful coronary stent implantation. The 2 groups were matched for clinical, angiographic, and procedure characteristics. RESULTS: No patients in the protamine group had adverse effects such as hypotension or vascular collapse during protamine administration. Acute stent thrombosis did not occur in any protamine group patient but did occur in 12 patients in the control group (0.7%) (P =.47). Subacute stent thrombosis occurred in 2 patients in the protamine group (2.1%) and in 15 in the control group (0.8%) (P =.20). By logistic regression analysis, protamine was not a determinant of stent thrombosis. CONCLUSIONS: Reversal of anticoagulation by protamine after stent implantation does not predispose to stent thrombosis. This result has important clinical consequences because it allows the use of protamine in the treatment of coronary perforation and serious bleeding complications that may occur during coronary stent deployment. PMID- 10385767 TI - Effect of clinical factors on length of stay after coronary artery bypass surgery: results of the cooperative cardiovascular project. AB - BACKGROUND: Rising health care costs have prompted careful review of comparative hospital resource use. Length of stay after bypass surgery has received particular attention. However, many providers assert that these variations are caused by differences in the clinical mix of patients treated. Our goals were to identify the major clinical predictors of postoperative length of stay (PLOS) after coronary artery bypass graft surgery (CABG), document variations in PLOS among 28 hospitals, and assess the degree to which patient characteristics account for hospital variations in PLOS. METHODS: Detailed clinical data on 3605 Medicare patients undergoing CABG in 28 Alabama and Iowa hospitals were analyzed by stepwise linear regression to identify significant clinical predictors of PLOS. Analysis of variance was used to compare hospitals' PLOS while controlling for significant patient risk factors. RESULTS: The mean age was 72.1 years, 34.7% were female, and the in-hospital mortality rate was 5.6%. The median and mean PLOS were 8 and 11.1 days, respectively. Significant predictors of longer PLOS included increasing age, female sex, history of chronic obstructive pulmonary disease, cerebrovascular disease, or mitral valve disease, elevated admission blood urea nitrogen, and preoperative placement of an intraaortic balloon pump. Hospitals varied significantly (P =.0001) in their unadjusted PLOS. These hospital-level variations persisted despite adjustment for both preoperative patient characteristics (P =.0001) and postoperative complications and death (P =.0001). CONCLUSIONS: This study found significant between-hospital variations in PLOS that were not explained by patient factors. This finding suggests the potential for increased efficiency in the care of patients undergoing CABG at many institutions. Further research is needed to determine the practice patterns contributing to variations in length of stay after bypass surgery. PMID- 10385768 TI - Continuous intravenous dobutamine is associated with an increased risk of death in patients with advanced heart failure: insights from the Flolan International Randomized Survival Trial (FIRST). AB - OBJECTIVE: To evaluate clinical characteristics and outcomes of patients with advanced heart failure receiving intravenous continuous dobutamine in the FIRST Trial (Flolan International Randomized Survival Trial). METHODS: Four hundred seventy-one patients with class IIIb to IV heart failure who were enrolled in the FIRST trial were included. Eighty patients treated with dobutamine at FIRST randomization were compared with 391 patients not treated with dobutamine at randomization. The occurrence of worsening heart failure, need for vasoactive medications, resuscitated cardiac arrest, myocardial infarction, and total mortality were compared between the 2 groups. RESULTS: The dobutamine group had a higher occurrence of first event (85.3% vs 64. 5%; P =.0006) and a higher mortality rate (70.5% vs 37.1%; P =.0001) compared with the no dobutamine group. Intravenous continuous dobutamine was an independent risk factor for death after adjusting for baseline differences. CONCLUSIONS: Dobutamine use at the time of randomization was associated with a higher 6-month mortality rate. This effect persisted after adjustment for baseline differences. This analysis challenges the concept that continuous intravenous dobutamine is beneficial to advanced heart failure patients with respect to survival. PMID- 10385769 TI - Insights into the contemporary epidemiology and outpatient management of congestive heart failure. AB - OBJECTIVES: To evaluate the epidemiology, prognosis, and patterns of practice in patients with chronic congestive heart failure (CHF) treated and followed at a specialized clinic. METHODS: Prospective cohort study of consecutive patients referred to and followed up in a specialized heart failure clinic between September 1989 and March 1996. RESULTS: Of the 628 patients referred, 566 were confirmed to have CHF. Mean duration of follow-up was 518 +/- 490 days (range 1 to 2192 days). Vital status was available for 99.3% of patients. Mean age at enrollment was 66 years, 68% were men, 67% had an ischemic cause of heart disease, and 78% had systolic dysfunction. Patients with preserved systolic function were older, more often female, had higher mean systolic blood pressures, and a lower prevalence of ischemic heart disease, ventricular arrhythmias, or impaired renal function when compared with those with systolic dysfunction (all P 45% (P =.04). There was also a negative correlation between cardiac troponin T and LVEF (R = -0.41, P =.01). CONCLUSIONS: These data show that cardiac troponin T is increased in patients with congestive heart failure and that the level parallels the severity of the disease. We conclude that cardiac troponin T is a suitable candidate-marker molecule to monitor congestive heart failure from a structural perspective. PMID- 10385771 TI - Self-reported differences between cardiologists and heart failure specialists in the management of chronic heart failure. AB - BACKGROUND: Heart failure (HF) is responsible for considerable mortality morbidity rates and resource utilization. Recently, several studies have reported improved outcomes when patients are managed by special HF clinics, but it is uncertain whether this improvement reflects differences in physician practices or other aspects of the operation of these clinics. OBJECTIVES: This study was designed to identify differences in HF management practices between general cardiologists and cardiologists specializing in the treatment of patients with HF. METHODS: A survey examining diagnostic and treatment practices in patients with HF was sent to a sample of cardiologists derived from the American Medical Association Masterfile and to HF specialists who were members of the Society of Transplant Cardiologists or principal investigators in HF trials. Responses were examined in relation to guidelines issued by the Agency for Health care Policy and Research released 9 months previously. RESULTS: In general both groups practice in conformity with published guidelines. However, there were important differences between the practice patterns of general cardiologists and HF specialists. For instance, in patients being evaluated for the first time, cardiologists reported using a chest radiograph to assist in the diagnosis more than did HF specialists (47% vs 12%), whereas HF specialists were more likely to use an echocardiogram (73% vs 48%). Both groups were likely to evaluate their patients for ischemia and possible revascularization, even in patients not having angina. However, HF specialists tended to use coronary angiography as the initial diagnostic test, whereas cardiologists were more likely to use stress testing. HF specialists more often used angiotensin-converting enzyme inhibitors as part of their initial therapy in patients with mild to moderate HF (94% vs 86%) and during maintenance therapy (91% vs 80%). Also, HF specialists were more likely than cardiologists to titrate angiotensin-converting enzyme inhibitors to higher doses (75% vs 35%), even in the presence of renal dysfunction. CONCLUSION: Cardiologists and HF specialists generally manage their patients in conformity with guidelines. However, in many areas, such as angiotensin-converting enzyme inhibitor use, HF specialists do so more aggressively. These approaches may, in part, explain the success of the HF clinic model and raise the possibility that some portion of the HF population may be more optimally managed by cardiologists with a special interest in and additional training or experience with this condition. PMID- 10385772 TI - Actual failure rates: A method of assessing tissue valve reoperation rates. AB - BACKGROUND: Patients older than age 70 with coronary disease undergoing mitral valve replacement have a limited survival rate, suggesting that few of these patients will require reoperation if they receive a tissue mitral valve. However, traditional actuarial analysis of valve durability censors patients who die before valve failure and doesn't reflect the lower frequency of reoperation in patient subgroups with limited survival rates. METHODS: Actual or cumulative reoperation rates represent the rates of reoperation in patients without censoring for death and estimate the risk that the valve will fail before the patient dies. The actual rates of valve rereplacement were determined in 255 recipients of tissue mitral valves, categorizing patients by age and presence of coronary artery disease at 8 years after surgery and compared with standard actuarial estimates of valve durability. RESULTS: Operative mortality rates were 17.5% for patients with coronary artery disease and 6.0% for those without (P <.001). At 8 years, the actual reoperation rates for patients younger than age 70 with and without coronary artery disease were 9.2% (n = 76) and 10% (n = 90), respectively. In patients older than age 70 without coronary artery disease the reoperation rate was 9.4% (n = 32). In contrast, the actual reoperation rate was only 1.7% (n = 58) in patients older than age 70 with coronary artery disease. This rate was significantly lower (P =.05) than the other groups. The difference in reoperation rates was not significant if standard actuarial analysis was used. CONCLUSIONS: Actual reoperation rates are significantly lower in patients older than 70 years with CAD receiving mitral tissue valves than in younger patients or in patients without coronary artery disease. Calculation of actual reoperation rates provides a unique and clinically useful perspective in evaluating data on prosthetic valve reoperation rates. PMID- 10385773 TI - Complications of Inoue balloon mitral commissurotomy: impact of operator experience and evolving technique. AB - BACKGROUND: There have been no single-center studies that have systematically addressed the acute outcome of Inoue balloon mitral commissurotomy (BMC) performed in a large series of patients. Accordingly, this study sought to examine the impact of operator experience and continuing technical modifications on the success and complication rates of BMC. METHODS: BMC was performed in 799 patients: 469 patients with pliable mitral valves (group 1) and 330 patients with calcified valves and/or severe subvalvular disease (group 2). Acute complications were examined and compared between groups before and after modifications in BMC techniques. Major modifications included the use of a height-derived balloon sizing method for the selection of an appropriate balloon catheter, a cautionary stepwise dilation technique, and avoidance of traction on the interatrial septum during balloon inflations. RESULTS: Technical failures were encountered in 4 (0.5%) patients in our early experience. One patient sustained cardiac perforation and tamponade and was the only case requiring emergency surgery. There were no deaths. Systemic embolic events were observed in 11 (1.4%), all among the first 353 patients before the routine use of pre-BMC transesophageal echocardiography. Severe postprocedure angiographic (>/=3+) mitral regurgitation occurred in 4% of patients, 2% in group 1 versus 9% in group 2 (P =.0001). With increased operator experience and technical modifications, this complication was significantly reduced from 5% (7 of 150 patients) to 0% in the last 316 patients in group 1 (P =.0001) and from 11% (26 of 228 patients) to 3% (3 of 101 patients) in group 2 (P =.031). The incidence of significant interatrial shunting (pulmonary-to-systemic flow ratio >/=1.3) was also significantly reduced from 12% to 6% (P =.0034). CONCLUSION: Incremental operator experience and ongoing technical refinements in BMC techniques have resulted in a 100% technical success rate and a significant diminution in complications in patients with a wide spectrum of stenotic mitral valve morphologic features. PMID- 10385775 TI - Electrophysiologic procedures and activation of the hemostatic system. AB - BACKGROUND: Thromboembolism occurs in 0.4% to 2% of the subjects undergoing radiofrequency ablation (RFA), but its mechanisms remain unclear. Our aim was to evaluate several parameters of the hemostatic system in relation to the electrophysiologic procedure. METHODS: Thirty consecutive patients were enrolled in the study. Fifteen underwent electrophysiologic study and 15 underwent radiofrequency ablation. Before the ablation procedure, all subjects were given an intravenous heparin bolus (2500 IU). Blood samples were drawn immediately before, at the end of, and 24 hours after the procedures. Spontaneous platelet aggregation in whole blood and in platelet-rich plasma, markers of clotting activation (prothrombin fragment 1+2 and the thrombin-antithrombin complex) and the fibrinolytic system (plasminogen activator inhibitor and D-dimer) levels were evaluated. RESULTS: At the end of the procedure, spontaneous platelet aggregation in whole blood, prothrombin fragment 1+2, thrombin-antithrombin complex, and D dimer levels increased significantly in all patients. The hemostatic changes were more marked after RFA than after electrophysiology. Spontaneous aggregation in whole blood, prothrombin fragment 1+2, and thrombin-antithrombin complex levels at 24 hours after the procedure were similar to those observed before the procedure in both groups; D -dimer levels were still elevated with respect to preprocedure levels, with a trend toward higher levels in patients undergoing RFA rather than electrophysiology. A significantly more marked activation of coagulation (prothrombin fragment 1+2, P <.005) was found in patients in whom the mean duration of energy application was higher than 23.5 seconds. CONCLUSIONS: Our data suggest that antithrombotic prevention with a prolonged administration of heparin and/or the association of antiplatelet agents should be considered in patients undergoing RFA. PMID- 10385774 TI - Association of fasting blood sugar level, insulin level, and obesity with left ventricular mass in healthy children and adolescents: The Bogalusa Heart Study. AB - BACKGROUND: Insulin resistance, often associated with obesity, is hypothesized to be involved in the pathogenesis of essential hypertension and may relate to increased left ventricular mass (LVM). METHODS: We examined correlations between echocardiographic LVM and fasting blood glucose and insulin levels in a cross section of 216 black and white healthy children and young adults aged 13 to 27 years in Bogalusa, Louisiana. Anthropometric measurements and blood pressure readings were also obtained. RESULTS: Positive bivariate correlation was found between fasting blood glucose level and LVM corrected for growth (LVMC) (LVMC = LVM/Height2.7) with all race/sex groups combined (r = 0.17, P 8 seconds) was observed in 41% of patients with 60 series leads compared with only 11% with dedicated bipolar leads (P <.01). No significant effects on redetection were noted with an integrated lead with greater spacing between the tip and coil (70 series). CONCLUSIONS: Delayed redetection is frequently noted with an integrated lead with close spacing between the tip and coil. Detailed evaluation of detection and redetection of these leads should be performed at the time of pulse generator replacement. PMID- 10385777 TI - Alternate-day dosing of aspirin in atrial fibrillation. LASAF Pilot Study Group. AB - BACKGROUND: In inhibiting platelet function, aspirin seems to reduce the risk of cerebrovascular accidents, death, and acute coronary events in patients with nonrheumatic atrial fibrillation. Aspirin given on alternate days might have the advantage of not hindering prostacyclin synthesis. Thus a study was performed to evaluate whether aspirin used in this way might improve the results reported with daily treatment. METHODS: To test this hypothesis 285 patients (age range 40 to 82 years) with primary atrial fibrillation were randomly allocated in an open multicenter trial to 3 groups: (1) group A1, treated with 125 mg aspirin daily (n = 104); (2) group A2, treated with 125 mg aspirin on alternate days (n = 90), (3) group C (controls), who were not treated with anticoagulants or platelet inhibitors (n = 91). RESULTS: Inclusion took place from January 1990 to July 1994, and follow-up ended in May 1996 (range 1 to 62 months). Sudden cardiac death in association with heart failure or angina was the most common final event: 4.8%, 1.1%, and 6.6% in groups A1, A2, and C, respectively. Both cardiovascular mortality rate and the incidence of main events were reduced, in relative terms, by 80% (1. 1% in group A2 vs 6.6% in group C). The differences were smaller between group A1 and C but did not reach statistical significance. The reduction of main cardiovascular events between groups A1 and A2 was statistically significant (7.7% vs 2.2% = 5.5%; 95% confidence limits 1%, 11%; P <.05). The difference did not reach statistical significance when other end points were analyzed. CONCLUSION: In this trial low-dose aspirin given on alternate days seemed to be an efficient intervention in preventing major cardiovascular events. Regarding strokes, however, aspirin was less efficient. Mortality rate in the 3 groups as a whole was associated with heart failure and the development of ischemic heart disease. PMID- 10385778 TI - Prophylactic oral amiodarone compared with placebo for prevention of atrial fibrillation after coronary artery bypass surgery. AB - BACKGROUND: Postoperative atrial fibrillation occurs in 20% to 40% of patients undergoing coronary artery bypass grafting (CABG) and contributes to delayed recovery, increased length of stay, and increased hospital cost. Measures at preventing postoperative atrial fibrillation have had mixed results. We report a double-blind trial comparing oral amiodarone with placebo for the prevention of atrial fibrillation after CABG. METHODS AND RESULTS: All patients undergoing CABG were considered eligible. Exclusion criteria included bradycardia (<50 beats/min), prior Atrial fibrillation, concurrent therapy with antiarrhythmic drugs, or concomitant valve surgery. Patients were given 2 g of amiodarone (73 patients) or placebo (70 patients) in divided doses 1 to 4 days before surgery and 400 mg daily for 7 days postoperatively. Atrial fibrillation occurred in 24.7% (18 of 43) of patients receiving amiodarone and 32. 8% (23 of 70) of patients receiving placebo (P =.30). Heart rate at onset of atrial fibrillation was 133.4 +/- 26.6 beats/min for amiodarone compared with 152.9 +/- 31.6 beats/min for placebo (P =. 04). Duration of atrial fibrillation was 10.2 +/- 8.1 hours for amiodarone compared with 16.2 +/- 27.5 hours for placebo (P =.67). Patients receiving both beta-blockade and amiodarone had a 16.7% incidence of atrial fibrillation compared with 31.9% in the remaining patients (P =.10). Atrial fibrillation was associated with an increased cost of $7011 compared with those who remained in sinus rhythm ($23,869 +/- $20,894 vs $16,857 +/- $5401 in sinus rhythm). Hospital cost of those taking amiodarone was $18,895 +/- $13,267 compared with $18,839 +/- $11,537.18 for placebo (P =.42). CONCLUSION: Postoperative CABG atrial fibrillation is associated with prolonged hospital stay and increased cost. Prophylactic oral amiodarone did not statistically alter the incidence or duration of atrial fibrillation after CABG, although favorable trends were noted. Hospital cost was not affected by therapy with amiodarone. PMID- 10385779 TI - Long-term safety and efficacy of combination gemfibrozil and HMG-CoA reductase inhibitors for the treatment of mixed lipid disorders. AB - BACKGROUND: Combinations of gemfibrozil and a 3-hydroxy-3-methylglutaryl (HMG) coenzyme A reductase inhibitor show promise in treating mixed lipid abnormalities. However, concern regarding the risk of myopathy and hepatic toxicity has limited the use of this combination. To determine the long-term safety and efficacy of this combination, we prospectively identified all patients placed on a combination of gemfibrozil and any HMG reductase inhibitor. METHODS: Pravastatin, simvastatin, fluvastatin, lovastatin, or atorvastatin at incremental doses was combined with gemfibrozil (600 mg twice daily). Lipid profiles, creatine kinase levels, and aminotransferase levels were monitored. Two hundred fifty-two patients with established atherosclerosis receiving combination therapy for a mean of 2.36 +/- 1.52 years spanning a total of 593.6 patient-years were monitored. RESULTS: In 148 patients, gemfibrozil was started before an HMG was added. The pretreatment total cholesterol level fell from 222 +/- 34 mg/dL to 181 +/- 26 mg/dL (P <.001) on combination therapy. HDL cholesterol level rose from 30 +/- 5 mg/dL to 36 +/- 7 mg/dL (P <.01), triglyceride level fell from 361 +/- 141 mg/dL to 212 +/- 101 mg/dL (P <.03). The ratio of total cholesterol to HDL fell from 7.6 +/- 1. 7 to 5.3 +/- 1.6 (P <.001). In 104 patients an HMG was begun before gemfibrozil was added. Pretreatment total cholesterol level fell from 246 +/- 54 mg/dL to 192 +/- 40 mg/dL on combination therapy (P <.01). HDL level rose from 33 +/- 9 mg/dL to 38 +/- 9 mg/dL (P <.03) and triglyceride level fell from 314 +/- 183 mg/dL to 183 +/- 93 mg/dL (P <.001). The ratio of total cholesterol to HDL fell from 7.9 +/- 3.6 to 5.2 +/- 1.4 (P <.001). In both groups the lipid profile on combination therapy was significantly better than that obtained on single-agent therapy. One episode of myopathy (0.4%) and one episode of aminotransferase level elevation (0.4%) of greater than 3 times upper limit of normal occurred. Both resolved with cessation of therapy without consequence. CONCLUSIONS: Combinations of gemfibrozil and an HMG, compared with either agent alone, results in improved long-term control of lipid abnormalities in mixed lipid disorders. The low incidence of toxicity permits the use of combination therapy in patients at high risk of atherosclerotic complications. PMID- 10385780 TI - Comparison of the hypolipidemic effect of gemfibrozil versus simvastatin in patients with type III hyperlipoproteinemia. AB - BACKGROUND: Type III hyperlipoproteinemia is characterized by the accumulation of chylomicron and very low density lipoprotein (VLDL) remnants. Individuals with this disorder have a high risk of premature atherosclerosis, and hypolipidemic drugs are useful in their management. METHODS: We compared, in a double-blind, placebo-controlled, randomized crossed study, the effects of gemfibrozil (1200 mg/day) and simvastatin (20 mg/day) on lipids, apolipoprotein AI, apolipoprotein B, and apolipoprotein E and on lipids and apolipoprotein B content in VLDL, intermediate-density lipoprotein (IDL), low-density lipoprotein (LDL), and high density lipoprotein (HDL) in 10 patients with type III hyperlipoproteinemia. RESULTS: Levels of total cholesterol, VLDL cholesterol, IDL cholesterol, and apolipoprotein B decreased with both drugs. Larger reductions in triglycerides (109 +/- 28.2 mg/dL, P =.005), VLDL cholesterol (24.7 +/- 10.9 mg/dL, P =.05), and VLDL triglycerides (86.3 +/- 20.2 mg/dL, P =.003) were obtained with gemfibrozil compared with simvastatin. LDL cholesterol reduction was more effective with simvastatin than with gemfibrozil (44.3 +/- 17.1 mg/dL, P =.03). HDL cholesterol after gemfibrozil was 5.71 +/- 2.37 mg/dL higher than after simvastatin. CONCLUSIONS: In patients with type III hyperlipoproteinemia gemfibrozil is more effective in reducing total triglyceride and VLDL lipid levels than simvastatin, and simvastatin is better in reducing LDL cholesterol than gemfibrozil is. IDL and apolipoprotein E levels were reduced similarly with both drugs. PMID- 10385781 TI - Improved endocardial visualization with second harmonic imaging compared with fundamental two-dimensional echocardiographic imaging. AB - BACKGROUND: Endocardial visualization is suboptimal by fundamental imaging in at least 30% of patients. Second harmonic imaging was developed for visualization of myocardial contrast agents. We have hypothesized that endocardial visualization may improve with harmonic imaging compared with fundamental imaging. METHODS AND RESULTS: Accordingly, 40 consecutive patients with poor endocardial visualization by conventional echocardiography in at least 1 left ventricular segment (22 segment model) in the 4 standard views randomly underwent fundamental and harmonic imaging without contrast. The images were separately and randomly analyzed by 2 observers. Endocardial visualization was scored as 0, not visualized; 1, poorly visible; and 2, well visualized. Endocardial visualization indexes were also calculated. More segments were assigned a score of 0 (P <. 001) and 1 (P <.001) by fundamental compared with harmonic imaging, whereas harmonic imaging demonstrated more segments with a score of 2 (P <.001) compared with fundamental imaging. Endocardial visualization indexes were significantly better by harmonic imaging in the parasternal long axis (P <.005), short axis (P <.001), and apical 4- (P <.0001) and 2-chamber views (P <.0001). Similar results were obtained by a second observer. Agreement between the 2 observers regarding improvement, deterioration, or no change in score between harmonic and fundamental imaging was 88% (kappa = 0. 76). Interobserver and intraobserver agreements for systolic wall thickening scores also significantly improved with harmonic compared with fundamental imaging (P <.001). CONCLUSION: Second harmonic imaging is superior to fundamental imaging for endocardial visualization in patients with suboptimal fundamental imaging. PMID- 10385782 TI - Effects of endurance exercise training on left ventricular systolic performance and ventriculoarterial coupling in patients with coronary artery disease. AB - BACKGROUND: Endurance exercise training can increase left ventricular (LV) ejection fraction during dynamic exercise in coronary artery disease. This adaptation may be mediated by altered cardiac loading conditions rather than an improvement in intrinsic LV systolic function. To minimize these confounding effects, we used isometric handgrip exercise to assess the training-induced changes in LV systolic function and ventriculoarterial coupling. METHODS: Twenty six patients (52 +/- 2 years of age) trained for 12 months. LV function was assessed with radionuclide ventriculograpy. RESULTS: LV systolic reserve (the change in LV ejection fraction from rest to handgrip exercise) increased from 7.32 +/- 1.2 to -3.4 +/- 1.1 (P =. 033) without acute changes in end-diastolic volume or the effective arterial load. LV end-systolic elastance increased 37% (P =.039) during handgrip exercise. Resting end-diastolic volume increased and the effective arterial load decreased after training. CONCLUSIONS: Data suggest that in coronary artery disease adaptations to exercise training include a lower effective arterial load and an increase in EDV at rest, with an improvement in LV systolic function detectable only during afterload stress. PMID- 10385783 TI - Highlights from the American College of Cardiology 48th Annual Scientific Sessions: March 7 to 10, 1999. PMID- 10385785 TI - Risk stratifying acute coronary syndromes: gradient of risk and benefit. AB - The pathophysiology of acute coronary syndromes, ranging from unstable angina to non-Q-wave to Q-wave myocardial infarction (MI) has been elucidated over the past 2 to 3 decades. Treatments involving clot lysis and antiplatelet therapy have reduced the morbidity and mortality rates of these syndromes. In patients with persistently elevated ST segments, treatment with thrombolytic agents is well established. In patients with unstable angina and non-Q-wave MI, there is ongoing investigation of the use of antithrombins and antiplatelet agents. Unfortunately, these treatments do not come without risk. The use of cardiac troponins is currently under intensive investigation because of their specificity to cardiac muscle and sensitivity in the determination of minimal myocardial injury. Troponin T and troponin I are more sensitive than CK-MB and are likely able to detect microinfarctions in patients with unstable angina. Most important is the ability of troponin T and troponin I to identify patients with unstable angina without ST elevations who are at high risk for cardiac events, including MI and cardiac death. Early risk stratification in the emergency department with the cardiac troponins allows for more appropriate decisions for admission and therapeutic triage of admitted patients. One such use currently being studied is the ability of the troponins to identify patients who will benefit from glycoprotein IIb/IIIa receptor inhibitors. PMID- 10385784 TI - Potential non-glycoprotein IIb/IIIa effects of abciximab. AB - The antithrombotic effect of abciximab is believed to be primarily due to its blockade of platelet glycoprotein IIb/IIIa receptors, leading to the inhibition of platelet aggregation. Studies have, however, identified that antibody 7E3, the parent molecule of abciximab, and/or abciximab itself, binds to both "activated" alphaMbeta2 receptors and alphaVbeta3 receptors. Because alphaMbeta2 receptors are present on granulocytes and monocytes, cells that have been implicated in contributing to atherosclerosis, intimal hyperplasia after vascular injury, reperfusion injury, and thrombin generation, it is possible that some of abciximab's effects relate to this reactivity. Similarly, because alphaVbeta3 has been implicated in platelet adhesion to osteopontin, intimal hyperplasia after vascular injury, and platelet-mediated thrombin generation, it is possible that some of abciximab's beneficial effects relate to this reactivity. Blockade of alphaVbeta3 receptors may also be beneficial in other disease states because, in animal models, such blockade inhibits tumor angiogenesis and sickle cell adhesion to blood vessel endothelium. Despite these intriguing observations, there are no direct data to support any beneficial roles or any unwanted side effects related to the reactivities of abciximab with "activated" alphaMbeta2 or alphaVbeta3 receptors. PMID- 10385786 TI - Glycoprotein IIb/IIIa blockade and thrombolytics: early lessons from the SPEED and GUSTO IV trials. AB - Reperfusion with a regimen of thrombolytic therapy, aspirin, and unfractionated heparin is limited by a number of factors. Only 50% to 60% of patients achieve early thrombolysis in myocardial infarction grade-3 flow within 90 minutes with the most effective thrombolytic regimens. Even after initial reperfusion is achieved, transient and permanent reocclusion occurs too often and is associated with high mortality rates. As more older patients are treated, intracranial hemorrhage is becoming more common. Finally, the risk of bleeding and procedural failure has been high in patients who received an acute percutaneous interventional procedure shortly after treatment with thrombolytic therapy. Given the important role of platelets in the thrombotic process and the relatively weak inhibitory effect of aspirin, it is reasonable to seek agents that will provide more profound platelet inhibition. Early studies with full-dose fibrinolytic and glycoprotein IIb/IIIa inhibitors have been promising, but concern about bleeding has hindered this strategy. Several recent trials have evaluated full-dose abciximab with reduced-dose fibrinolytic therapy and have yielded promising results. PMID- 10385787 TI - Platelet glycoprotein IIb/IIIa antagonists in percutaneous coronary revascularization. AB - Coronary dissections and intracoronary thrombosis are the most important determinants of acute and subacute complications occurring during coronary interventions. Stents have proved efficacious to repair coronary dissections. The formation of intracoronary thrombi during percutaneous coronary intervention (PCI), the other main pathogenic mechanism for acute complications of PCI, has also been implicated in the pathogenesis of long-term ischemic outcomes. To reduce intracoronary thrombosis, all PCIs have been historically performed during intense short-term anticoagulation with the combination of unfractionated heparin plus aspirin. Compelling data on the central role of platelets in arterial thrombosis have made apparent the need for more potent antiplatelet agents for the treatment of patients undergoing PCI. PMID- 10385788 TI - Economic issues in glycoprotein IIb/IIIa receptor therapy. AB - Efficacy, safety, and cost will determine the use of glycoprotein IIb/IIIa therapy in patients with acute coronary syndromes or those patients undergoing percutaneous coronary intervention (PCI). Prospective randomized studies with abciximab, eptifibatide, and tirofiban have demonstrated the superior efficacy and relative safety of IIb/IIIa therapy in these 2 broad patient groups. In medical practice, we by necessity make decisions to administer or withhold therapies based on implicit concepts of cost-effectiveness and efficacy and safety. We herein review available economic data on IIb/IIIa therapy to assist in this decision-making process. The procurement costs of the IIb/IIIa receptor antagonists vary considerably for both acute coronary syndrome and patients undergoing PCI. In PCI, these procurement costs range from $436 to $1407 per patient treated with commonly used regimens. Economic substudies of PCI trials with abciximab and tirofiban demonstrate medical cost savings that partially offset drug procurement costs. The number of dollars spent on IIb/IIIa agents per death or myocardial infarction prevented in patients undergoing PCI ranges from $13,000 to $37,000. Abciximab has cost-effectiveness ratios of $4000 to $7000 per life-year saved in patients undergoing PCI. The incremental cost-effectiveness of IIb/IIIa blockade in the setting of planned stenting is unknown. In patients with acute coronary syndrome, procurement costs range from $1050 to $1548 per patient treated. Expenditures per death or myocardial infarction prevented in patients with acute coronary syndrome range from $32,000 to $82, 000. Inadequate direct cost data exist to calculate cost effectiveness ratios for this group, but only high-risk patients will likely have cost-effectiveness ratios that most Western health-care systems can afford. PMID- 10385789 TI - Readministration of abciximab: interim report of the ReoPro readministration registry. AB - Even with continued improvements in the technology of percutaneous coronary intervention (PCI), approximately 10% to 20% of patients undergoing PCI will require repeat procedures within 1 year. Furthermore, because of the chronic nature of coronary artery disease, many patients will require additional treatment with PCI well after an initial episode of care. Abciximab (ReoPro), a chimeric (murine/human) monoclonal antibody fragment (c7E3 Fab), has been shown to significantly improve periprocedural and long-term outcomes associated with PCI and to reduce the need for repeat target vessel revascularization. However, because the structure of abciximab is derived from an antibody, concern has been raised about subsequent repeat administration. To prospectively evaluate the safety and efficacy of abciximab readministration, we established the ReoPro Readministration Registry with the intent to determine the efficacy, human antichimeric antibody response and rates of thrombocytopenia, bleeding, intracranial hemorrhage, and anaphylaxis in at least 500 patients being retreated with abciximab. The study was conducted at 19 centers beginning in March 1997. This article details interim data that are based on the first 329 patients. Data to date indicate that readministration with abciximab is safe and efficacious and that the same indications for first-time use should apply to subsequent readministration. PMID- 10385791 TI - Letter PMID- 10385790 TI - Oral blockade of the platelet glycoprotein IIb/IIIa receptor: fact or fancy? AB - Currently available antiplatelet agents are limited in the scope and magnitude of platelet inhibition. Orally active platelet glycoprotein IIb/IIIa antagonists are currently in clinical testing. These agents may extend platelet inhibition and clinical benefit from parenteral glycoprotein IIb/IIIa antagonists. The most common adverse side effect is mucocutaneous bleeding, which is related to the magnitude and duration of platelet inhibition. Point-of-care monitoring of platelet function may enhance safety and efficacy of oral platelet GP IIb/IIIa blockade. Because aspirin, ticlopidine, and clopidogrel have proved beneficial in reducing vascular ischemic events, oral platelet glycoprotein IIb/IIIa inhibitors, which provide more marked platelet inhibition, are positioned to provide even greater clinical benefit if tolerated. PMID- 10385792 TI - Gastrointestinal toxicity is not a major complication of standard dose paclitaxel therapy. PMID- 10385793 TI - [Emergency contraception--the sooner the better]. PMID- 10385794 TI - [Parallel import of drugs]. PMID- 10385795 TI - [Can back pain be the cause of tender toes?]. PMID- 10385796 TI - [Rheumatoid arthritis]. PMID- 10385797 TI - [Well-educated and healthy]. PMID- 10385798 TI - [Social inequalities in cancer survival]. AB - Social differentials in survival from 12 common types of cancer are assessed by estimating an additive-multiplicative hazard model on the basis of individual register and census data for the whole Norwegian population. The excess all-cause mortality among cancer patients compared with similar persons without a cancer diagnosis is significantly related to education, occupation and income. It is, on the whole, about 15% lower for men or women with a completed post-secondary education than for those with only compulsory schooling, with age, period, histology and stage at the time of diagnosis taken into account. The prognosis is even better for those with an education at Master level, who can expect to live more than one year longer after the diagnosis than those with only compulsory education, as a rough average over the cancer sites. The data do not provide clear indications of whether differences in host factors, such as comorbidities, immune functions and lifestyle, or differences in treatment are primarily responsible for these inequalities in survival. PMID- 10385799 TI - [Pharmacists' and general practitioners' views on parallel import of drugs]. AB - Since 1995, parallel import of drugs to Norway has been allowed under the European Economic Area Agreement. The health authorities have stated that there are no concerns connected with the use of parallel-imported drugs. This study is an interview survey among general practitioners, pharmacy staff and pharmacists working in pharmacies. The findings show that 91% of the pharmacies dispense parallel-imported drugs and that there is a certain amount of scepticism regarding the use of parallel-imported drugs. Most respondents feel that parallel imported pharmaceuticals may have financial advantages for the individual patient and economic advantages for society at large, but 50% of the pharmacists and 54% of the physicians were of the opinion that parallel-imported pharmaceuticals represented an increased medical risk for the patients. Approximately 15% of doctors and pharmacy staff had knowledge of either incorrect treatment or adverse drug reactions due to the use of parallel-imported drugs. The time used for prescribing and dispensing parallel-imported drugs is longer than for directly imported preparations. The survey shows that approximately every fifth doctor will use the right of reservation in connection with the dispensing of parallel imported drugs, whereas approximately every fourth pharmacist will disregard the right of reservation. PMID- 10385800 TI - [Leg cramps in pregnancy--how common are they?]. AB - Leg cramps in pregnancy, defined as painful spasms of the calf, were investigated among women giving birth at the maternity ward at Baerum Hospital from 1 October to 20 October 1997. A questionnaire distributed to 120 women three days after parturition revealed that 45% had suffered from leg cramps during pregnancy. Among 54% of them the cramps appeared after the 25th week of pregnancy. 76% of the women had experienced the symptoms twice per week or less often; 81% of them suffered from painful cramps only during night-time. We conclude that leg cramps are still a common symptom in pregnancy and may compromize sleep and hence the ability to work. PMID- 10385801 TI - [X-linked recessive bulbospinal neuronopathy--Kennedy's syndrome]. AB - Kennedy's syndrome is an inherited disease which was probably first described 100 years ago. Although rare, a recent report suggests that the prevalence may show considerable regional differences. A review of 30 different names of the disease is given. Originally, the disorder was regarded as a spinal and bulbar muscular atrophy but it is now obvious that there is severe axonal degeneration, also of the sensory fibres with the pattern of a central-peripheral distal axonal neuropathy. This was also present in the two recognized cases presented here. The sensory symptoms develop slowly and it is suggested that a peripheral sprouting compensates for the loss not only of motor, but also sensory fibres. It is important to distinguish the disease from motor neuron diseases since the progression is slow and the expected life span is normal. The clinical presentation with facial palsy and perioral contraction-fasciculations is pathognomonic. However, demonstration of increased (CAG)n repeat size in the androgen receptor gene is diagnostic. A normal (CAG)n repeat size excludes the diagnosis, since the abnormal expansion is the only mutation associated with the disease. Other types of mutations in the androgen receptor gene lead to a different clinical presentation, testicular feminization. PMID- 10385802 TI - [Cerebral visual impairment in children]. AB - Visual handicaps in children are often difficult to diagnose in early age and several etiological possibilities have to be ruled out. Cerebral visual impairments are of greater importance than most epidemiological studies indicate. Children with low birth weight are at special risk for visual disturbancies. Children with developmental dysfunctions and retardation and with neurohandicaps may have combined anterior ocular pathology and cerebral visual impairment. Health professionals responsible for the investigation of children with learning disabilities and visuomotor problems, must have a good understanding of visual development and processing. PMID- 10385803 TI - [Atrial fibrillation--epidemiological and electrophysiological aspects]. AB - Atrial fibrillation is a common and therapy-requiring cardiac arrhythmia. The chronic form becomes identified in population studies from the age of 50 and increases with age. The number of Norwegian individuals with this arrhythmia is estimated to be slightly in excess of 40,000 and it will increase approximately by another 5,000 until the year 2010. Mechanisms responsible for initiation and maintenance of the arrhythmia are increasingly better understood. The paroxysmal form often has a focal origin, allowing curative treatment, but can also be associated with signs of deficient interatrial conduction. The chronic form is perpetuated partly by a shortening of atrial myocardial refractoriness, attributed to a failure in the intracellular turnover of Ca-ions due to the high excitation rate. Ongoing studies are expected to illustrate the clinical benefit of Ca-blockers prior to cardioversion of chronic atrial fibrillation. The further relapse rate is low in patients who have maintained sinus rhythm more than 1-2 months following conversion. It is therefore possible that, following a successful conversion to sinus rhythm, aggressive antiarrhythmic treatment should be given during a limited period only--a strategy which has to be evaluated in prospective studies. PMID- 10385804 TI - [An elderly woman with arthritis, liver disease and later muscular paralysis]. PMID- 10385805 TI - [Spinal disorders in Norway--an epidemiological report]. AB - Low back disorders are prevalent and induce large costs to the health services, the national insurance system and employers. This paper describes the prevalence of low back pain and low back work disability in the Norwegian population, and the incidence and duration of low back work disability in Aust-Agder county. Data from the Norwegian Health Survey 1995 and the registers of the National Insurance Administration are presented. 45% of women and 38% of men reported low back pain within a 14 day period. The prevalence of low back work disability was 1.9%. Low back disorders caused 13-17% of sickness absence, rehabilitation allowance and disability pensions in 1995. In Aust-Agder, the incidence of sickness absence caused by low back disorders was higher among men. In 16% of persons sick-listed due to low back pain without radiating symptoms, the sickness absence episode lasted for more than seven weeks. When radiating symptoms were present, the corresponding figure was 35%. The number of persons with low back disorders with radiating symptoms seems to have increased the most. Further research, preventive measures and guidelines for clinical work should focus on the group of persons who develop chronic low back work disability. PMID- 10385806 TI - [The pain takes hold of life. A qualitative study of how patients with chronic back pain experience and cope with their life situation]. AB - Living with daily pain during a long period of time is a considerable strain which affects many aspects of life. The aim of this study was to explore the experiences and coping strategies of patients with chronic low back pain. A strategic sample of 22 patients with chronic low back pain were interviewed according to a semistructured qualitative method: 14 women and eight men, with a mean age of 46 and a mean duration of illness of 15 years. 18 had been treated with low back surgery. All patients had pain every day. 19 were never without pain. They reported disturbed sleep, irritable mood and depression; 15 had thought of suicide. They felt constantly worn out. The pain influenced their everyday life and their total situation. The lives of patients with intermittent daily pain were less affected. The patients were asked what advice they would give to others in a similar situation. Their main point was not to give up, to focus less on the pain and the limitations and more on everything that could bring dedication and joy to everyday life. PMID- 10385808 TI - [The vision of zero traffic accidents in Norway]. PMID- 10385807 TI - [Being believed is what counts. A qualitative study of experiences with the health service among patients with chronic back pain]. AB - The article presents a qualitative study of patient experiences with the health service and their general practitioners. 22 patients with daily pain were interviewed. They represent a strategic sample of patients with chronic low back pain, but without paresis. Their statements were not as critical as those voiced in the public debate. 18 patients had a continuing patient-doctor relationship. They were all satisfied with their doctor. A need for more information and a better understanding of the patient was present in all interviews. Many of the patients stressed the importance of feeling that the doctor honestly understood their problems. In addition to practical help, the patients also wanted the opportunity to talk to the doctor about their difficult life situation, however without actually expecting that much could be done about it. The most important aspect was to be able to talk about their life situation with a competent person. Kindliness and thoughtful consideration from the doctor meant a lot to the patients in what they experienced as a very difficult life situation. PMID- 10385809 TI - [Spinal medicine--reliability of the specialty is suffering]. PMID- 10385810 TI - [Non-smoking environment is a human right]. PMID- 10385811 TI - [Serotonergic syndrome. Serotonergic syndrome caused by paroxetine]. PMID- 10385812 TI - [University function at the central hospital in Akershus]. PMID- 10385813 TI - [Villonodular synovitis--a rare cause of knee joint locking]. PMID- 10385814 TI - [Research philosophy and practical research--new knowledge about alendronate and fractures]. PMID- 10385815 TI - [On hemoglobin, venesection, statins and mortality]. PMID- 10385816 TI - [Tampons]. PMID- 10385818 TI - [Insufficient knowledge about Karl Evang]. PMID- 10385819 TI - [Contact lenses]. PMID- 10385820 TI - Effective immediately Injury Prevention expands to all age groups. PMID- 10385821 TI - Dumpers, rips, and other hazards for the environmental surfer. PMID- 10385822 TI - Reducing the burden of injury. PMID- 10385823 TI - A mother's tale: escalator amputates four fingers. PMID- 10385824 TI - One adolescent's injury--a continuing saga. PMID- 10385825 TI - My sister's death. PMID- 10385826 TI - Alcohol and other psychoactive drugs in trauma patients aged 10-14 years. AB - OBJECTIVE: To examine the prevalence of alcohol and/or other psychoactive drugs, such as marijuana and cocaine (AODs), involved in preteen trauma patients. METHODS: Toxicological testing results were analyzed for 1356 trauma patients aged 10-14 years recorded in the National Pediatric Trauma Registry for the years 1990-95. RESULTS: Of the 1356 patients who received toxicological screening at the time of admission, 116 (9%) were positive for AODs. AOD involvement increased with age. Patients with pre-existing mental disorders were nearly three times as likely as other patients to be AOD positive (23% v 8%, p < 0.01). AOD involvement was more prevalent in intentional injuries and in injuries that occurred at home. CONCLUSIONS: AODs in preteen trauma are of valid concern, in particular among patients with mental disorders or intentional injuries. The role of AODs in childhood injuries needs to be further examined using standard screening instruments and representative study samples. PMID- 10385827 TI - Unintentional injury mortality in children: a priority for middle income countries in the advanced stage of epidemiological transition. AB - OBJECTIVES: To examine the relationship between the magnitude, and the relative importance of unintentional child injury mortality with socioeconomic development, and to conceptualise the dynamic changes in injury mortality within the framework of epidemiological transition. DESIGN: Ecological cross sectional study using data on 51 countries. MAIN OUTCOME MEASURES: The relationship between total mortality rates, unintentional injury mortality rates, and percentage in children 1-14 years of age with gross national product (GNP) per capita. RESULTS: Unintentional injury mortality rates in children were negatively correlated with GNP per capita. However, by categorising the data, we found some areas of non correlation: in children 5-14 years in low income versus lower middle income countries, and in all age and gender groups in lower high income versus higher high income countries. A high percentage of total deaths due to injuries was clearest in the lower middle income countries in all age and gender groups. CONCLUSIONS: The changes in child injury mortality in relation to socioeconomic development could be conceptualised as three stages: a stage of high magnitude; a stage of high priority; and a stage of improvement. Most middle income countries are in the high priority stage where both injury mortality rates and injury percentage of total deaths are high. PMID- 10385828 TI - "First aid for scalds" campaign: reaching Sydney's Chinese, Vietnamese, and Arabic speaking communities. AB - OBJECTIVES: As a serious yet preventable problem, scald injuries in children have been a priority for prevention in Australia and other developed countries. Not only can the occurrence of scalds be prevented, but immediate first aid treatment offers an effective method for secondary prevention, reducing the severity of scalds. Despite the success of scald prevention initiatives, local evidence suggested that first aid knowledge was lacking in some minority ethnic groups. To redress this gap, the "First Aid for Scalds" campaign for those from a non English speaking background was specifically targeted to three ethnic groups (Vietnamese, Chinese, and Arabic), with the aim of increasing the proportions of parents and caregivers who had correct knowledge of first aid treatment for scalds. The primary strategy was a media campaign, including advertisements on ethnic radio and in ethnic newspapers. METHODS: The evaluation design included formative research and impact evaluation. The impact evaluation study involved random population based telephone surveys with each of the three language groups, before and after the campaign, to assess the reach and effectiveness of the campaign. RESULTS: After the campaign, there were significant increases in the proportion of people who knew the correct first aid treatment for scalds. There were substantial variations in campaign recall and knowledge between each of the three language groups. The largest improvement was found in the Vietnamese group. CONCLUSION: The association between campaign recall and increase in correct knowledge, and the absence of any similar interventions during the campaign period, give credence to the conclusion that the changes observed were a result of the campaign. The results demonstrate the value of community based injury prevention campaigns specifically targeting linguistically diverse communities. PMID- 10385829 TI - Evaluation of a drowning prevention campaign in King County, Washington. AB - OBJECTIVES: A three year drowning prevention campaign focused on increasing the use of life vests among children 1-14 years old. An evaluation was conducted to determine campaign awareness, change in ownership and use of life vests by children, and predictors of life vest use. SETTING: King County, Washington. METHODS: Four telephone surveys were conducted with parents before, during, and after the campaign. RESULTS: The campaign was recalled by 50% of families surveyed. From before to after the campaign, reported life vest use by children on docks, beaches, or at pools increased from 20% to 29% (p < 0.01) and life vest ownership for children increased from 69% to 75% (p = 0.06). Among parents aware of the campaign, reported child life vest use increased from 20% to 34% (p < 0.001) and ownership increased from 69% to 80% (p < 0.01). Among families unaware of the campaign, neither life vest use nor ownership changed significantly. Children were more often reported to wear life vests if a parent knew of the campaign, was confident fitting the vest, was younger than 40 years, felt the child could not swim well, and owned a life vest for the child. CONCLUSIONS: A community-wide drowning prevention campaign resulted in a significant, although modest, increase in reported life vest use and ownership among children. PMID- 10385830 TI - Achieving compliance with pool fencing legislation in New Zealand: a survey of regulatory authorities. AB - OBJECTIVES: To identify the status of compliance and enforcement of New Zealand's Fencing of Swimming Pools Act (FOSP Act), 10 years after its introduction, and to identify methods for improving both compliance with the act and the process of enforcement. METHODS: A postal questionnaire was sent to all 74 authorities in New Zealand in which they were asked questions about their enforcement of the FOSP Act. Semistructured telephone interviews were conducted with 12 authorities to supplement the data obtained in the postal survey. RESULTS: Based on responses to the survey, it was estimated that there are over 59,000 domestic swimming pools in New Zealand, giving rates of 46 pools/1000 dwellings and 16 pools/1000 persons. The authorities reported that 44% of pools complied with the act, and a further 4% had been granted exemptions. Nineteen per cent of pools were reported to not comply with the act, and the compliance status of a further 33% was not known, or not stated by the authority. Only 9% of authorities had procedures for locating and inspecting pools, while 28% had a programme of reinspection to ensure that pools continued to comply. Pool owner resistance was considered to be the main difficulty with enforcing the act, and nearly half of the authorities believed publicity or education was needed to overcome these barriers. Fifty two per cent of authorities had publicized the act during the 12 months preceding the survey. CONCLUSIONS: Due to ambiguities within the legislation, and differing levels of commitment by authorities to locate pools and monitor compliance, compliance with the FOSP Act is not consistent nationally. If the act were less ambiguous, there would be greater consistency and more enforcement. PMID- 10385831 TI - Injury hospitalizations among American Indian youth in Washington. AB - OBJECTIVE: To determine the rate and causes of hospitalizations for injury among American Indian and Alaska Native (AI/AN) youth in the state of Washington, and to compare this with the rate of hospitalizations for injury among youth of all races. METHODS: Subjects were aged 0-19 years and were admitted to civilian hospitals for care of an injury (International Classification of Diseases N codes 800-995) in Washington between 1990 and 1994. Deaths occurring in the prehospital setting and emergency department are not included. Using several fields of identifying information, the Washington state hospital discharge database was linked with the Indian Health Service (IHS) patient registration database to identify AI/AN youth. Denominator data included the total age specific IHS user population for American Indians and US Census derived population estimates. Incidence ratios (IRs) were calculated to compare rates of hospitalization between AI/AN youth and all youth in Washington. RESULTS: A total of 694 and 29,048 hospitalizations for injury were identified for AI/AN youth and all races, respectively. The rate of hospitalization for injuries among AI/AN youth was 507 discharges per 100,000 youth (IR = 1.30; 95% confidence interval (CI) 1.20 to 1.40. The leading mechanism of injury was motor vehicles (IR 1.73, CI 1.49 to 2.01), followed by falls (IR 0.95, CI 0.79 to 1.15), and poisoning (IR 1.20, CI 0.80 to 1.78). The disparity was greater for intentional injuries (IR 1.71, CI 1.44 to 2.04). The highest IR for all unintentional injuries was for injuries from fire (IR 2.35, CI 1.42 to 3.87). AI/AN children aged 15-19 had the greatest disparity for rates of injury hospitalization (IR 1.4, CI 1.25 to 1.56). CONCLUSION: AI/AN youth in Washington had a higher hospitalization rate for injury compared with all youth in the state. Disparities were greatest for injuries related to motor vehicles and assaults. When linked, hospital discharge data can be used for surveillance of AI/AN hospitalizations. PMID- 10385832 TI - An intervention to reduce playground equipment hazards. AB - OBJECTIVES: A community intervention trial was carried out to evaluate the relative effectiveness of two methods of reducing playground hazards in schools. The study hypotheses were: (1) a health promotion programme addressing barriers to implementing the New Zealand Playground Safety Standard will reduce playground hazards and (2) the intervention programme will be more successful than providing information alone. METHODS: Twenty four schools in Wellington, New Zealand were randomly allocated into two groups of 12 and their playgrounds audited for hazards. After the audit, the intervention group received a health promotion programme consisting of information about the hazards, an engineer's report, regular contact and encouragement to act on the report, and assistance in obtaining funding. The control group only received information about hazards in their playground. RESULTS: After 19 months, there was a significant fall in hazards in the intervention schools compared with the control schools (Mann Whitney U test, p = 0.027). No intervention schools had increased hazards and eight out of 12 had reduced them by at least three. In contrast, only two of the control schools had reduced their hazards by this amount, with three others increasing their hazards in that time. CONCLUSIONS: It is concluded that working intensively with schools to overcome barriers to upgrading playground equipment can lead to a reduction in hazards, and that this form of intensive intervention is more effective than providing information alone. PMID- 10385833 TI - Children's fractures: a population based study. AB - OBJECTIVE: To measure the incidence of childhood fractures in a defined population. SETTING: Accident and emergency (A&E) departments covering the Swansea and Neath Port Talbot areas of South Wales in 1996. METHODS: Linkage of data from A&E departments with population data to produce fracture incidence rates by anatomical site and cause in children aged 0-14 years. RESULTS: During 1996, 2463 new fractures occurred in 2399 residents yielding a fracture rate of 36.1/1000 children. Fractures were more common in boys than girls and increased with age in both groups. Sports and leisure activities accounted for 36% of fractures, assaults for 3.5%, and road traffic accidents 1.4%. Fractures of the radius/ulna were most frequent (36%). CONCLUSIONS: The fracture rate in South Wales children is twice the rate reported in previous studies. Further research is required to elucidate the reasons behind this high rate. Many fractures could be prevented by the use of safer surfaces in school playgrounds, and wrist protection in in-line skaters and possibly in soccer players. PMID- 10385834 TI - Graduated licensing comes to the United States. AB - OBJECTIVE: To describe the young driver problem and the emergence of graduated licensing as a way to address it. METHODS: Literature review and commentary. RESULTS: Twenty-four states in the United States adopted versions of graduated licensing in 1996-98; initial results show positive effects. CONCLUSIONS: A major public health movement is under way that can be expected to produce significant reductions in crashes and injuries involving young drivers. PMID- 10385835 TI - Comparing pediatric intentional injury surveillance data with data from publicly available sources: consequences for a public health response to violence. AB - OBJECTIVE: A hospital based intentional injury surveillance system for youth (aged 3-18) was compared with other publicly available sources of information on youth violence. The comparison addressed whether locally conducted surveillance provides data that are sufficiently more complete, detailed, and timely that clinicians and public health practitioners interested in youth violence prevention would find surveillance worth conducting. SETTING: The Boston Emergency Department Surveillance (BEDS) project was conducted at Boston Medical Center and the Children's Hospital, Boston. METHOD: MEDLINE and other databases were searched for data sources that report separate data for youth and data on intentional injury. Sources that met these criteria (one national and three local) were then compared with BEDS data. Comparisons were made in the following categories: age, gender, victim-offender relationship, injury circumstance, geographic location, weapon rates, and violent injury rates. RESULTS: Of 14 sources dealing with violence, only four met inclusion criteria. Each source provided useful breakdowns for age and gender; however, only the BEDS data were able to demonstrate that 32.6% of intentional injuries occurred among youth aged 12 and under. Comparison data sources provided less detail regarding the victim offender relationship, injury circumstance, and weapon use. Comparison of violent injury rates showed the difficulties for practitioners estimating intentional injury from sources based on arrest data, crime victim data, or weapon related injury. CONCLUSIONS: Comparison suggests that surveillance is more complete, detailed, and timely than publicly available sources of data. Clinicians and public health practitioners should consider developing similar systems. PMID- 10385836 TI - Validity of self reported crashes and injuries in a longitudinal study of young adults. AB - OBJECTIVES: The aim of this study was to determine the validity of self report as a source of information on crashes and injuries. SETTING: This study was part of the Dunedin Multidisciplinary Health and Development Study (DMHDS), which is a longitudinal study of the health, development, and behaviour of a cohort of young New Zealanders. METHOD: At the age 21 assessment DMHDS study members were asked to report serious injury and motor vehicle traffic crashes experienced over the previous three years. The self reported injuries were compared with the New Zealand Health Information Service (NZHIS) public hospital discharge file to determine the completeness of the self reported data. The traffic crashes were compared with the police traffic crash reports to determine the accuracy of self reported crash details. RESULTS: Twenty five (86%) of the 29 unintentional injuries, six (67%) of the nine assaults, and one (14%) of the six self inflicted injuries on the NZHIS file were self reported. The level of agreement between the self reported crash details and those recorded on the traffic crash report was high. CONCLUSIONS: The results show that self reports can be a useful and valid source of injury and crash data. PMID- 10385837 TI - House fire injury prevention update. Part I. A review of risk factors for fatal and non-fatal house fire injury. AB - OBJECTIVE: To summarize house fire injury risk factor data, using relative risk estimation as a uniform method of comparison. METHODS: Residential fire risk factor studies were identified as follows: MEDLINE (1983 to March 1997) was searched using the keywords fire*/burn*, with etiology/cause*, prevention, epidemiology, and smoke detector* or alarm*. ERIC (1966 to March 1997) and PSYCLIT (1974 to June 1997) were searched by the above keywords, as well as safety, skills, education, and training. Other sources included: references of retrieved publications, review articles, and injury prevention books; Injury Prevention journal hand search; government documents; and internet sources. When not provided by the authors, relative risk (RR), odds ratio, and standardized mortality ratios were calculated, to enhance comparison between studies. RESULTS: Fifteen relevant articles were retrieved, including two case-control studies. Non modifiable risk factors included young age (RR 1.8-7.5), old age (RR 2.6-3.6), male gender (RR 1.4-2.9), non-white race (RR 1.3-15.0), low income (RR 3.4), disability (RR 2.5-6.5), and late night/early morning occurrence (RR 4.1). Modifiable risk factors included place of residence (RR 2.1-4.2), type of residence (RR 1.7-10.5), smoking (RR 1.5 to 7.7), and alcohol use (RR 0.7-7.5). Mobile homes and homes with fewer safety features, such as a smoke detector or a telephone, presented a higher risk of fatal injury. CONCLUSIONS: Risk factor data should be used to assist in the development, targeting, and evaluation of preventive strategies. Development of a series of quantitative systematic reviews could synthesize existing data in areas such as house fire injury prevention. PMID- 10385838 TI - Handguns as a pediatric problem. 1986. AB - Handgun injury is a major cause of morbidity and mortality in American society, particularly for young people. Large numbers of children are affected by handgun violence through the loss of fathers, brothers, and other relatives. Young children are injured and sometimes killed in handgun accidents. Some children and many adolescents are murdered with handguns. Because of their great lethality and very limited ability to provide personal protection, the great burden of handgun injury can best be reduced by making handguns less available. Handgun control cannot reduce rates of crime or interpersonal assault, but it can be expected to reduce the frequency and severity of injury which grows out of these situations, to levels closer to the much lower ones found in other countries. Pediatricians can contribute to this effort, as they have to the efforts to reduce the morbidity and mortality from poisonings and motor vehicle passenger injury. PMID- 10385839 TI - From theory to practice. PMID- 10385840 TI - Behavioral and electrophysiologic predictors of treatment response to stimulants in children with attention disorders. AB - Behavioral and quantitative electroencephalography (EEG) techniques were used to evaluate treatment response to stimulant therapy in children with attention disorders. A sample of 130 children with attention disorders were evaluated with Conners and Diagnostic and Statistical Manual of Mental Disorders--III rating scales, and with neurometric quantitative EEG before and 6 to 14 months after treatment with stimulants. Significant quantitative EEG differences were found between the normal control population (N = 31) and the children with attention problems. Quantitative EEG abnormalities involved increased theta or alpha power, greatest in frontal regions, frontal theta/alpha hypercoherence, and posterior interhemispheric power asymmetry. Behavioral improvement after stimulant treatment was seen in 81.5% of the children with attention-deficit hyperactivity disorder and 44.7% of the children with attention-deficit disorder, with the degree of correspondence between behavioral and quantitative EEG changes at 78.5%. Pretreatment clinical and quantitative EEG features could predict treatment response with a sensitivity of 83.1% and a specificity of 88.2%. A combined behavioral and quantitative EEG approach can be useful for following and predicting treatment response to stimulants in children with attention disorders. PMID- 10385841 TI - Spontaneous partial regression of low-grade glioma in children with neurofibromatosis-1: a real possibility. AB - At the age of 41 and 31 months, respectively, a boy and a girl affected by neurofibromatosis-1 were diagnosed with a visual pathway glioma during surveillance contrast-enhanced head magnetic resonance imaging (MRI). In the first child, the initial MRI showed that the entire optic chiasm, the intracranial tract of the left optic nerve, and hypothalamus were grossly enlarged and enhanced in the post-gadolinium T1-weighted images. Ten months later, the hypothalamic component of the lesion had regressed markedly and there were no more areas of contrast enhancement. In the second child, the initial MRI showed that the optic chiasm, the right optic tract, and geniculate body were enlarged and enhanced after gadolinium injection. At 6-month follow-up, the MRI showed that the right optic tract and the anterior aspect of the optic chiasm decreased in size and the contrast enhancement of the entire lesion was reduced dramatically. These findings, as indicated by other similar reports, confirm that spontaneous regression of visual pathway glioma is a rare but real possibility in children with neurofibromatosis-1. Therefore, clinicians need to be aware of visual pathway glioma's erratic behavior in children with neurofibromatosis-1 with special attention given to the importance of a very conservative attitude toward any type of treatment for such patients. PMID- 10385842 TI - Carbamazepine withdrawal in children with previous symptomatic partial epilepsy: effects on neuropsychologic function. AB - Neurocognitive performance was evaluated in seven children with symptomatic partial epilepsy prior to, and at least 12 months after, discontinuation of carbamazepine. The patients treated with carbamazepine monotherapy were seizure free for at least 2 years and without electroencephalographic anomalies for at least 1 year. Results indicated that carbamazepine at therapeutic levels does not affect intellectual, memory, or attentional functions, or more complex frontal functions. Nevertheless, after therapy withdrawal scores on frontal function tests used in this study improved significantly. This suggests that these functions could have been better without carbamazepine therapy. The fact that carbamazepine decreases neuron membrane excitability and could reduce the information circuity, particularly in the frontal areas, is offered as a possible explanation. Further studies on larger samples using the same design are required to validate these results. PMID- 10385843 TI - Reflex sympathetic dystrophy in children. AB - Reflex sympathetic dystrophy is a syndrome characterized by pain in one or more extremities, usually associated with vasomotor changes. Its occurrence in childhood has long been thought to be rare. We describe six cases of pediatric reflex sympathetic dystrophy and suggest that this syndrome could be underdiagnosed in children and adolescents. Psychologic problems frequently play a role in this disorder, which often can be treated conservatively. We also point out that the diagnosis is mainly clinical. An early diagnosis can avoid unnecessary tests and potentially can improve response to treatment, and prognosis. PMID- 10385844 TI - Molar tooth sign in Joubert syndrome: clinical, radiologic, and pathologic significance. AB - Joubert syndrome is a rare autosomal-recessive condition characterized by early hyperpnea and apnea, developmental delay, and truncal ataxia. We previously described key ocular motor signs in Joubert syndrome and the molar tooth sign resulting from dysplasia of the isthmic segment of the brain stem, superior cerebellar peduncles, and vermis. In this study, we obtained clinical and developmental data in 61 cases, and radiologic data in 46 of these, to determine the prevalence of the molar tooth sign in a large sample, and to ensure that magnetic resonance images obtained for study were representative of the Joubert syndrome population at large. We studied the morphology of the isthmic segment of the pontomesencephalic junction, the segment of the brain stem derived from the primitive isthmus. Portions of the cerebellum analyzed included the superior cerebellar peduncles, the anterior and posterior lobes of the vermis, and the flocculonodular lobe. In one case, autopsy of the brain was performed. The average age at diagnosis was 33 months. All patients were hypotonic and developmentally delayed. The molar tooth sign was present in 85% of cases with 13% of these showing additional malformations. All patients without the molar tooth sign had other mimicking conditions such as neocerebellar dysgenesis, isolated vermian atrophy, cerebellar aplasia, and cystic dilation of the cisterna magna. Autopsy showed aplasia of the cerebellar vermis with dysplasia of the dentate nucleus, elongated locus coeruleus, and marked dysplasia of the caudal medulla. A better understanding of the clinical, radiologic, and pathologic features of Joubert syndrome should help uncover the genetic basis for the syndrome. PMID- 10385845 TI - Expression of transforming growth factor-beta 1 in periventricular leukomalacia. AB - The expression of transforming growth factor-beta 1, which has neurotrophic effects, was investigated in 25 neonates with periventricular leukomalacia using immunohistochemistry. In controls, transforming growth factor-beta 1 immunoreactivity was not detected in the cerebral white matter or cortex. Of the 25 cases of periventricular leukomalacia, transforming growth factor-beta 1 immunoreactivity was found in 16, and was distributed mainly in the cytoplasm of astrocytes, being prominent around necrotic foci in the white matter. The immunoreactivity was negative or weak at the acute stage of periventricular leukomalacia, and increased at the subacute stage and then decreased or was absent at the chronic stage. Astrocytes that were moderately or markedly positive for transforming growth factor-beta 1 were not found before 27 weeks' gestation, but were observed after 32 weeks' gestation in the white matter of the brains of neonates with periventricular leukomalacia. Transforming growth factor-beta 1 expression tended to be more obvious in focal periventricular leukomalacia than in widespread or diffuse periventricular leukomalacia. Our results suggest that transforming growth factor-beta 1 could be involved in the delayed glial response rather than the initial glial activation, and could play a role in the inhibition and repair of injury in periventricular leukomalacia. Exogenous transforming growth factor-beta 1 could have therapeutic potential for periventricular leukomalacia. PMID- 10385846 TI - Racial differences in free radical scavenging enzyme activity in children. AB - Oxygen-derived free radicals play an important role in multiple pediatric neurologic diseases. Five intracellular free radical scavenging enzymes and three trace elements provide a significant portion of the body's defenses against free radical-mediated injury. Although the effects of age, sex, and ethnicity on the body's antioxidant defenses have been described, no study has examined whether racial differences exist. This pilot study sought to determine the effect of racial differences on the activity of five free radical scavenging enzymes and the concentrations of three associated trace elements in normal, healthy American children. The erythrocyte and plasma activities of five major free radical scavenging enzymes (glutathione peroxidase, glutathione reductase, glutathione-S transferase, catalase, and superoxide dismutase) and plasma concentrations of three associated trace elements (selenium, copper, and zinc) were determined for 83 healthy American children, aged 1 to 18 years. One- and two-way interactions of race, age, and sex with each dependent variable were analyzed. African Americans had higher erythrocyte glutathione peroxidase activity, erythrocyte superoxide dismutase activity, and selenium and copper concentrations than Caucasians. Racial inequalities do exist in free radical scavenging enzyme activity and trace element concentration in healthy children. African-American children had higher activity in the two most important free radical scavenging enzymes used by the brain compared to age- and sex-matched Caucasian children. Future clinical research in free radical-mediated pediatric neurologic diseases needs to consider race along with age and sex in both study design and data analysis. PMID- 10385847 TI - Familial clustering of autoimmune disorders and evaluation of medical risk factors in autism. AB - Autism is an age-dependent neurologic disorder that is often associated with autoimmune disorders in the patients' relatives. To evaluate the frequency of autoimmune disorders, as well as various prenatal and postnatal events in autism, we surveyed the families of 61 autistic patients and 46 healthy controls using questionnaires. The mean number of autoimmune disorders was greater in families with autism; 46% had two or more members with autoimmune disorders. As the number of family members with autoimmune disorders increased from one to three, the risk of autism was greater, with an odds ratio that increased from 1.9 to 5.5, respectively. In mothers and first-degree relatives of autistic children, there were more autoimmune disorders (16% and 21%) as compared to controls (2% and 4%), with odds ratios of 8.8 and 6.0, respectively. The most common autoimmune disorders in both groups were type 1 diabetes, adult rheumatoid arthritis, hypothyroidism, and systemic lupus erythematosus. Forty-six percent of the autism group reported having relatives with rheumatoid diseases, as compared to 26% of the controls. Prenatal maternal urinary tract, upper respiratory, and vaginal infections; asphyxia; prematurity, and seizures were more common in the autistic group, although the differences were not significant. Thirty-nine percent of the controls, but only 11% of the autistic, group, reported allergies. An increased number of autoimmune disorders suggests that in some families with autism, immune dysfunction could interact with various environmental factors to play a role in autism pathogenesis. PMID- 10385848 TI - Ethosuximide is effective in the treatment of epileptic negative myoclonus in childhood partial epilepsy. AB - The aim of our study was to evaluate the effectiveness of ethosuximide in the treatment of epileptic negative myoclonus, a motor disorder that can occur in childhood partial epilepsy. We introduced ethosuximide in nine patients with partial epilepsy of varying etiology (idiopathic, cryptogenic, symptomatic) who presented with epileptic negative myoclonus. The drug was added to the patients' preexisting antiepileptic drugs, which were maintained unchanged for the following 6 months. Epileptic negative myoclonus disappeared in all patients 15 to 30 days after ethosuximide was started. Plasma ethosuximide levels ranged from 55 to 89 micrograms/mL. The clinical response was not influenced by the patients' preexisting treatment or by the etiology of the epilepsy. No side effects were observed, and none of the patients presented a recurrence of epileptic negative myoclonus during follow-up. Furthermore, in five patients we observed the disappearance of partial seizures; in the remaining patients seizures were reduced by more than 75%. Electroencephalograms showed a decrement or disappearance of focal paroxysmal abnormalities. Our results suggest that ethosuximide is effective in the treatment of epileptic negative myoclonus and that it should be considered as a first-choice drug in the treatment of this motor disorder. PMID- 10385849 TI - Tuberous Sclerosis Consensus Conference: recommendations for diagnostic evaluation. National Tuberous Sclerosis Association. AB - At the recent Tuberous Sclerosis Consensus Conference, a subcommittee proposed recommendations to guide the rational use of diagnostic studies in patients with tuberous sclerosis complex. Recommendations were made for diagnostic evaluation at the time of diagnosis, when testing helps both to establish the diagnosis and to identify potential complications. Additional guidelines were proposed for the ongoing surveillance of established patients to detect later complications of tuberous sclerosis complex. In the absence of comprehensive population studies to govern the use of diagnostic studies in individuals with tuberous sclerosis complex, the panel developed guidelines based on the disorder's natural history, concentrating on complications that are common, clinically significant, and more easily managed when found early. Finally, the group made suggestions for the use of diagnostic tests to identify family members who have tuberous sclerosis complex. Although these recommendations should standardize and improve our use of diagnostic studies in individuals with tuberous sclerosis complex, the clinical approach in a given patient must remain flexible enough to meet the needs of individual patients and families. PMID- 10385851 TI - Monitoring with our good senses. PMID- 10385852 TI - Electronic availability of anesthesia records. PMID- 10385850 TI - Plasma baclofen levels in children receiving continuous intrathecal baclofen infusion. AB - To determine the plasma baclofen concentrations of children undergoing continuous intrathecal baclofen infusion for treatment of cerebral spasticity, we assayed plasma samples from six children, 8 to 18 years of age, who were receiving intrathecal baclofen at constant rates of 77 to 400 micrograms/day. Plasma levels were at or below the limit of quantification (10 ng/mL) in all patients. PMID- 10385853 TI - Stabilization and stability of twitch force during mechanomyography of the adductor pollicis muscle. AB - OBJECTIVE: In order to study the stabilization time, the increase in twitch force during stabilization and the maintenance of stability during mechanomyography of the adductor pollicis muscle, neuromuscular function was monitored in 20 patients anaesthetized without the use of a neuromuscular blocking agent. The effect of the type of stimulation (single twitch [ST; 0.1 Hz], or train-of-four [TOF; 4 stimuli at 2 Hz, repeated every 12 s]) on these variables was studied. When applying TOF stimulation, the variables were also investigated for the TOF percentage [quotient of fourth and first twitch of a TOF stimulus x 100%]. METHODS: Two groups of ten patients were monitored with either ST or TOF stimulation. Measurements continued for at least 45 minutes. Multiple linear regression analysis was applied to examine the effect of stimulation on the stabilization time and the increase in twitch force. RESULTS: According to our criteria for stability, we found that the stabilization time did not differ for ST (13.7 [10.2] min; mean [sd]) and TOF stimulation (18.1 [9.6] min) (p > 0.10). Stabilized twitch forces were larger during TOF than during ST stimulation (134% [19] and 113% [11]; p = 0.01). In both groups of stimulation, six patients showed an interruption of stability. The TOF percentage was stable throughout the measurement period in all patients. CONCLUSIONS: Stabilization of twitch force takes too long for many studies of neuromuscular function in the clinical research setting. Therefore, we do not recommend its routine use when performing mechanomyography of the adductor pollicis muscle. PMID- 10385854 TI - A method for assessment of standards of care of anesthesia services in departments with different levels of resources. AB - In developing countries the standards of anesthesia care vary greatly between hospitals. In order to identify the urgent needs of disadvantaged hospitals, we compared three index hospitals in the greater Cairo area, one of which has excellent (category I), on intermediate (category II), and one with severely restricted resources (category III). Standards of care published by the American Society of Anesthesiologists (ASA) were used to develop a spreadsheet for documenting features of pre-, intra- and post-anesthetic care in patients undergoing tonsillectomies, a procedure commonly performed in all three hospitals. The spreadsheet enabled us to document all equipment, supplies and personnel engaged from pre-anesthetic evaluation to discharge. Analysis of the data revealed that the service provided by the category I hospital approached the ASA standards. In the category II hospital the patients did not go through a pre anesthetic evaluation; instead they were seen for the first time in the operating room. No premedication was given. Intravenous access was established with the help of a needle (rather than a catheter). Monitoring consisted occasionally of a finger on the pulse. Sterilization was accomplished by boiling. Air-conditioning was not available. No records were kept and no recovery room was available. The same deficiencies existed in the category III hospital, which did not even have oropharyngeal airways, antiarrhythmic or inotropic medications, and sterile techniques were completely ignored. Despite these stark differences in care, the patients or their parents in all three hospitals appeared satisfied with the level of care they received. Much has to be done to improve anesthesia care in less fortunate departments in developing nations. Urgent help does not mean the need for sophisticated monitors or equipment only, but the establishment of practice standards first. Applying the priciples of modern management, we need to evaluate the structure, processes and outcome of anesthetic practice in developing countries in order to reengineer the way we provide help to anesthetic departments in developing nations. In this modest study we are presenting a means to evaluate the features and processes of the anesthesia services in developing countries. PMID- 10385855 TI - Perioperative tissue thickness measurement by a new miniature ultrasound device. AB - INTRODUCTION: A recently developed mini ultrasound device for measurement of peripheral tissue thickness is now available for use in clinical practice. Whether this device allows a better guidance of perioperative fluid therapy has to be investigated. Therefore, it is necessary to get basic data on the parameter tissue thickness in otherwise healthy patients during surgery. The aim of the present study was to evaluate differences in tissue thickness change between patients in supine and head down position with a novel handheld ultrasound device during the perioperative course of healthy surgical patients under a standardized fluid regimen. METHODS: After obtaining ethics committee approval and informed consent we studied 19 ASA 1-2 female patients undergoing gynecological procedures in supine (SUP, n = 11) or in 30 degrees head down position (HD, n = 8) in general anesthesia. Preoperative NPO status was comparable in both groups. Lactated Ringer's solution (LR) was continuously infused at a rate of 8 ml/kg b.w./h over 90 min and tissue thickness (TT) was determined by ultrasound before induction (t0) and in 30 min intervals (t30, t60, t90) at the forehead. Simultaneously plasma viscosity (PV) was evaluated. RESULTS: Group SUP presented at t0 a forehead TT of 5.3 mm (SD +/- 0.5), at t30 TT was unchanged. At t60 mean TT increased significantly to 5.6 mm, (+/- 0.6). At t90 mean TT remained stable at 5.7 mm (+/- 0.5). Group HD presented at t0 a mean TT of 4.6 mm (+/- 0.7), at t30 mean TT was 4.9 mm (+/- 0.7) and at t60 mean TT of the forehead skin was measured as 5.3 mm (+/- 0.6). Significance to t0 was reached at t90 with a mean TT of 5.4 mm (+/- 0.7). Group HD showed a steeper increase and a parallel stabilization phase at the end. Differences between t0 and t90 have been significant. Mean PV in the SUP group at t0 (1.361 mPa*s, SD: +/- 0.045) decreased under the infusion therapy to 1.276 mPa*s (+/- 0.04) at t90. Mean PV in the HD group was determined 1.351 mPa*s (+/- 0.06) at t0 and declined to 1.274 mPa*s (+/- 0.03) at t90. CONCLUSIONS: The findings suggest that fluid replacement after an NPO period and the expected changes of forehead TT due to positioning of the patient are detectable by this new ultrasound device. PMID- 10385856 TI - Measurement of blood concentration of indocyanine green by pulse dye densitometry -comparison with the conventional spectrophotometric method. AB - OBJECTIVE: Pulse dye densitometry (PDD) uses two wavelengths (805 and 890 nm) in association with pulse oximetry to compute the arterial blood concentration ratio of indocyanine green (ICG) to hemoglobin (Hb). When Hb is measured in the usual way, this permits the PDD to compute cardiac output, plasma or blood volume, and liver blood flow following an intravenous injection of ICG. In this study, we evaluate the accuracy of the PDD calculation of dye concentration by comparing it with measurement of the dye concentration in blood (Cb) measured by the spectrophotometric cuvette method during dye clearance in patients. METHODS: In 25 patients receiving major abdominal surgery, ICG (10, 20, or 40 mg) was injected into a central vein and arterial ICG concentration was continuously and simultaneously monitored at nose and finger by PDD; concurrently, ICG concentrations were measured by a spectrophotometer at 805 nm in 4 radial arterial blood samples. Repeated measures or one-way ANOVA were used for comparison of ICG concentrations and percent errors by PDD at the nose, finger, and Cb. RESULTS: The percent error (bias) of calculated dye concentration and its standard deviation (precision) was -3.9 +/- 16.8% (p < 0.01) with the probe on a nostril and 3.4 +/- 12.6% using the finger probe. These errors were found to be greatest when the mean transit time of the dye was rapid (-20.7 +/- 6.8% at nose p < 0.01 and -8.5 +/- 2.5% at finger p < 0.05) due to factors other than the time delay of blood sampling. CONCLUSION: These errors are of similar size to those associated with thermal cardiac output measurement, suggesting that PDD should be valuable clinically as a noninvasive tool especially since it provides values for blood volume and liver blood flow. PMID- 10385857 TI - The fast flush test--is the clinical comparison equivalent to its in vitro simulation? AB - OBJECTIVE: An in vitro simulation of the fast flush (FL) test has previously been used to prove that the FL test-measures the dynamic response of entire the blood pressure monitoring system. This simulation has also been used to confirm that the FL test is equivalent to the "gold standard" test for determining dynamic response, namely the square wave (SW) test. The conditions of the in vitro simulation can be reproduced in vivo during cardiopulmonary bypass (CPB) and circulatory arrest. Therefore the present objective was to verify that the previous conclusions about the validity of the FL test, obtained from an in vitro model, are equally valid when applied to in vivo clinical conditions. A secondary objective was to determine whether the patient's arterial tree has any affect on the dynamic characteristics of fluid-filled manometers. METHODS: Fourteen patients were studied during surgery that required CPB. We measured the dynamic response of the fluid filled arterial manometer during pulsatile conditions prior to the initiation of CPB, and then repeated the measurements during non-pulsatile CPB. In four of the fourteen patients we measured the dynamic response during circulatory arrest. A manometer, consisting of a fluid-filled tubing component, measured the patient's arterial blood pressure as well as the damped sinusoidal wave form created by the fast flush tests. The fluid-filled tubing was connected to a transducer (Utah Medical Products, Inc., Midvale, UT). The arterial pressures and the results of flush testing were recorded and displayed by a monitor (Marquette 7010, Marquette Electronics Inc., Milwaukee, WI). In an additional three patients we measured the dynamic response of the manometer in vitro and then in vivo. RESULTS: The dynamic response of the arterial pressure measuring system was the same during normal pulsatile flow, CPB and circulatory arrest. In addition, the dynamic response of the fluid-filled manometer was the same in vivo as in vitro. CONCLUSIONS: The clinical conditions during CPB and particularly during circulatory arrest duplicate the in vitro FL test simulation model. These results confirm the validity of the FL test in vivo as well as proving that the dynamic characteristics of a fluid-filled manometer are independent of the patient's vasculature. PMID- 10385859 TI - Use of medical simulators subject of international study. PMID- 10385858 TI - Use of an automated anesthesia information system to determine reference limits for vital signs during cesarean section. AB - INTRODUCTION: We evaluated whether automated anesthesia information systems can be used to calculate reference limits (population-based "normal values") for vital signs. We considered four populations of women undergoing cesarean section: healthy under spinal anesthesia, healthy under general anesthesia, pre eclamptic/eclamptic under spinal anesthesia, and pre-eclamptic/eclamptic under general anesthesia. METHODS: Reference limits were calculated for each of the study populations by determination of percentiles for: minimum heart rate, maximum heart rate, minimum arterial oxyhemoglobin saturation (SaO2), minimum mean arterial pressure (MAP), maximum MAP, decrease in MAP, and increase in MAP. RESULTS: There was one adverse anesthetic outcome among the 1,300 women in the study; the woman sustained a post-dural puncture headache. The 5th percentiles of SaO2 were at least 95% saturation under spinal versus 90% under general. Under spinal anesthesia, 95th percentiles for decreases in MAP from baseline were 63 mmHg for healthy and 75 mmHg for pre-eclamptic/eclamptic women. Under general anesthesia, the 95th percentiles for maximum MAP were 161 and 177 mmHg, respectively. Two women of the 1,300 patients experienced simultaneously a minimum SaO2 < 92% and minimum MAP < 50 mmHg. DISCUSSION: Automated anesthesia information systems can be used to determine reference limits for vital signs during anesthesia. Reference limits may play a role in malpractice cases when an expert claims that care by an anesthesiologist was sub-standard as shown by vital signs that were not maintained within the normal range during the critical portions of an anesthetic. Automated anesthesia information systems may enhance expert witnesses' clinical judgment. PMID- 10385860 TI - Low molecular weight heparin and epidurals. PMID- 10385861 TI - What is your diagnosis? Synovial sarcoma. PMID- 10385862 TI - Hypofractionated radiation therapy for invasive thyroid carcinoma in dogs: a retrospective analysis of survival. AB - Thirteen dogs with invasive thyroid carcinoma (WHO classification T2b or T3b) seen between January 1991 and October 1997 were treated by external beam irradiation. Four once-weekly fractions of 9 gray of 4 MeV X-rays were administered. Four of the dogs died of progression of the primary disease and four from metastatic spread. Of the remaining dogs, three died of unrelated problems, although two were still alive at the time of the censor. Kaplan-Meier analysis of the survival time from first dose to death from either primary or metastatic disease gave a median survival time of 96 weeks (mean 85 weeks, range six to 247 weeks). Radiographic evidence of pulmonary metastatic disease at presentation had no prognostic value whereas crude growth rate was a highly significant factor. The present series indicates that radiation therapy should be considered an important modality for the control of invasive thyroid carcinoma in the dog. PMID- 10385863 TI - Ultrasonographic appearance of primary gastric neoplasia in 21 dogs. AB - The ultrasonographic findings in 21 dogs with histologically confirmed primary gastric neoplasia were reviewed. Location, shape of the gastric lesion, evidence of gastric wall thickening, wall layers affected, presence of ulceration, evidence of extension through the gastric wall and lymphadenopathy were recorded. Twelve dogs with carcinoma shared many ultrasonographic features with six dogs that had lymphoma, the majority having sessile masses that appeared to involve all layers of the gastric wall; many also had evidence of ulceration and lymphadenopathy. Signs of extension of the lesion through the serosal surface of the stomach were identified ultrasonographically only in dogs with carcinoma. In contrast, three dogs with leiomyoma or leiomyosarcoma each had a focal mass affecting the gastric antrum, and lymphadenopathy was not identified ultrasonographically in these dogs. Even without any specific patient preparation, ultrasonography enables a morphological assessment of gastric neoplasms that may prompt a tentative diagnosis of gastric neoplasia and stimulate further investigation. PMID- 10385864 TI - Intestinal leiomyosarcoma in a cat. AB - Intramural ileocaecocolic leiomyosarcoma is described in an elderly neutered male domestic shorthaired cat with poor appetite and weight loss. Histological examination of the resected lesion revealed a poorly differentiated soft tissue sarcoma and a diagnosis of leiomyosarcoma was confirmed by positive immunohistochemical staining for vimentin and a-smooth muscle actin. Postoperative survival time was 102 days before local recurrence justified euthanasia. PMID- 10385865 TI - Leporacarus gibbus and Spilopsyllus cuniculi infestation in a pet rabbit. AB - A one-year-old male, chequered giant rabbit had a simultaneous infestation with both Leporacarus gibbus and Spilopsyllus cuniculi. Recurrent episodes of mild to severe pruritus had been noted over a period of two months. On clinical examination, partial alopecia and slightly erythematous skin with flea faeces was evident, although microscopic and cultural examinations of skin scrapings were negative for fungi. Parasitological examination, including adhesive tape strips of the rabbit's skin and fur, revealed L gibbus surface dwelling mites and the rabbit flea S cuniculi. The rabbit was successfully treated against both parasites with topical pyrethrin applied three times over a three-week period, and the clinical signs resolved in four weeks. PMID- 10385866 TI - Rectal prolapse associated with urinary bladder neoplasia in a cat. AB - A case of feline rectal prolapse which appeared to be secondary to transitional cell carcinoma of the urinary bladder is described. The cat was reported to be incontinent and treatment was declined by the owners. Euthanasia was performed and necropsy revealed an extensive nodular thickening of the entire urinary bladder wall. A diagnosis of urinary bladder transitional cell carcinoma was made on histopathological examination. PMID- 10385867 TI - Bilateral hydroureter and hydronephrosis in a nine-year-old female German shepherd dog. AB - A nine-year-old female German shepherd dog was presented in severe renal failure. Clinical and ultrasonographic examination revealed the presence of adrenal neoplasia, bilateral hydroureter and hydronephrosis but no evidence of urolithiasis or bladder neoplasia. In the absence of anuria, therapy for the renal failure was attempted but the azotaemia did not improve. Remarkably, bilateral hydroureter appeared to have been induced by a past routine surgical procedure--ovariohysterectomy. PMID- 10385868 TI - Immune-mediated thrombocytopenia associated with Angiostrongylus vasorum infection in a dog. AB - A three-year-old weimaraner was presented with lethargy, anorexia, neck pain and a soft fluctuant swelling in the thoracic inlet. A cough had been noted previously. Clinical examination revealed tachycardia, tachypnoea, pallor and a large subcutaneous swelling, with bruising, suggestive of a haematoma in the thoracic inlet. Thoracic radiographs revealed a cranial mediastinal mass which had the ultrasonographic appearance of fluid, and there was also a marked generalised interstitial lung pattern. Routine haematology revealed severe anaemia and thrombocytopenia, although coagulation tests were within normal limits. A diagnosis of immune-mediated thrombocytopenia was however made on the basis of a positive antiplatelet antibody test and a rapid response to prednisolone therapy. Furthermore, a tentative diagnosis of Angiostrongylus vasorum infection was suggested on the basis of clinical and radiographic findings, although no lungworm larvae were identified on faecal analysis. Despite initiating treatment with fenbendazole, the dog died suddenly. Postmortem examination revealed myocarditis, thrombosing arteritis, pneumonia and chronic membranoproliferative glomerulonephritis associated with A vasorum infection. PMID- 10385869 TI - Pathological fracture of the femur secondary to haematogenous osteomyelitis in a weimaraner. AB - A seven-year-old male weimaraner developed an acute-onset non-weightbearing pelvic limb lameness without a history of significant trauma. Radiographs demonstrated an oblique fracture of the femur associated with a lytic bone lesion, while cytology and histopathology of the lesion were consistent with osteomyelitis. A pure growth of Streptococcus intermedius was obtained from culture of affected tissue samples. The fracture healed without complication after rigid internal fixation and antibiotic therapy. PMID- 10385871 TI - Auditory processing disorders in children. PMID- 10385872 TI - Differential diagnosis and management of central auditory processing disorder and attention deficit hyperactivity disorder. AB - Children diagnosed with attention deficit hyperactivity disorder (ADHD) frequently present difficulties performing tasks that challenge the central auditory nervous system. The relationship between ADHD and central auditory processing disorder (CAPD) is examined from the perspectives of cognitive neuroscience, audiology, and neuropsychology. The accumulating evidence provides a basis for the overlapping clinical profiles yet differentiates CAPD and ADHD as clinically distinct entities. Common and distinctive management strategies are outlined. PMID- 10385873 TI - Acoustic-phonetic approach toward understanding neural processes and speech perception. AB - This review paper describes an "acoustic-phonetic" experimental approach aimed at understanding normal and abnormal speech perception processes from both a behavioral and an electrophysiologic perspective. First, we consider the relevant acoustic characteristics of speech and identify a set of acoustic-phonetic classes that represent the parameters most important for making an acoustic signal sound like speech. Second, we review what is known about the neurophysiologic representation of acoustic-phonetic speech parameters in animal and human subjects. Third, we describe how an acoustic-phonetic approach has been useful in understanding the biologic basis of some auditory learning problems in children and in characterizing the behavioral and neurophysiologic changes resulting from speech-sound training. Finally, we discuss these findings and how they may expand the diagnostic and rehabilitative repertoire of practicing audiologists. PMID- 10385874 TI - Multidimensional approach to the differential diagnosis of central auditory processing disorders in children. AB - Central auditory processing disorder (CAPD) may be viewed as a multidimensional entity with far-reaching communicative, educational, and psychosocial implications for which differential diagnosis not only is possible but also is essential to an understanding of its impact and to the development of efficacious, deficit-specific management plans. This paper begins with a description of some behavioral central auditory assessment tools in current clinical use. Four case studies illustrate the utility of these tools in clarifying the nature of auditory difficulties. Appropriate treatment options that flow logically from the diagnoses are given in each case. The heterogeneity of the population presenting with auditory processing problems, not unexpected based on this model, is made clear, as is the clinical utility of central auditory tests in the transdisciplinary assessment and management of children's language and learning difficulties. PMID- 10385875 TI - Habilitation and management of auditory processing disorders: overview of selected procedures. AB - This article describes three management approaches that can be used with children with auditory processing difficulties and learning disabilities. These approaches were selected because they can be applied in a variety of settings by a variety of professionals, as well as interested parents. The vocabulary building procedure is one that potentially can increase the ability to learn new words but also can provide training on contextual derivation of information, which is key to auditory closure processes. This procedure also helps increase language base, which can also enhance closure abilities. Auditory memory enhancement is a simple technique that involves many complex brain processes. This procedure reduces detailed information to a more gestalt representation and also integrates the motor and spatial processes of the brain. This, in turn, more fully uses working memory and helps in formulization and recall of important concepts of the sensory input. Finally, several informal auditory training techniques are discussed that can be readily employed in the school or home setting. These auditory training techniques are those that are most relevant to the kinds of deficits most often observed in our clinic. PMID- 10385876 TI - Auditory gap detection, perceptual channels, and temporal resolution in speech perception. AB - This article overviews some recent advances in our understanding of temporal processes in auditory perception. It begins with the premise that hearing is the online perceptual elaboration of acoustic events distributed in time. It examines studies of gap detection for two reasons: first, to probe the temporal acuity of auditory perception in its own right and, second, to show how studies of gap detection have provided new insights into the processes involved in speech perception and into the architecture of auditory spatial perceptual mechanisms. The implications of these new data for our comprehension of some central auditory processing disorders are examined. PMID- 10385877 TI - Molecular epidemiology in cancer research. AB - The use of molecular biomarkers in epidemiological investigations brings clear advantages of economy, speed and precision. Epidemiology--the study of the factors that control the patterns of incidence of disease--normally requires large numbers of subjects and/or long periods of time, because what is measured (the occurrence of disease) is a rare event. Biomarkers are measurable biological parameters that reflect, in some way, an individual's risk of disease-because they indicate exposure to a causative (or protective) agent, or because they represent an early stage in development of the disease, or because they allow an assessment of individual susceptibility. Biomarkers must be usable on one of the few materials available for biomonitoring of humans, i.e. blood, urine, exfoliated epithelial cells and, with some difficulty, biopsies. The approach of molecular epidemiology has a great potential is several areas of cancer research: investigating the aetiology of the disease; monitoring cancer risk in people exposed to occupational or environmental carcinogens; studying factors that protect from cancer; and assessing intrinsic factors that might predispose to cancer. The biomarkers most commonly employed in cancer epidemiology include: measurements of DNA damage--DNA breaks, altered bases, bulky adducts--in lymphocytes; the surrogate marker of chemical modifications to blood proteins, caused by agents that also damage DNA; the presence of metabolites of DNA damaging agents (or the products of DNA damage themselves) in urine; chromosome alterations, including translocations, micronuclei and sister chromatid exchange, resulting from DNA damage; mutations in marker genes; DNA repair; and the differential expression of a variety of enzymes, involved in both activation and detoxification of carcinogens, that help to determine individual susceptibility. The molecular approach has been enthusiastically employed in several studies of occupational/environmental exposure to carcinogens. While the estimation of biological markers of exposure has certainly shown the expected effects in terms of DNA damage and adducts, the detection of the biological effects of exposure (e.g. at the level of chromosome alterations) has not been so clear-cut. This is true also when smokers are examined as a group compared with non-smokers. Several markers (especially of chromosome damage and mutation) show a strong correlation with age-indicating either an increasing susceptibility to damage with age, or an accumulation of long-lived changes. DNA repair--a crucial player in the removal of damage before it can cause mutation--may vary between individuals, and may be modulated by intrinsic or extrinsic factors, but limited data are available because of the lack of a reliable assay. Information on other enzymes determining individual susceptibility does exist, and some significant effects of phenotypic or genotypic polymorphisms have emerged, although the interactions between various enzymes make the situation very complex. The important question of whether oxidative DNA damage in normal cells is decreased by dietary antioxidants (vitamin C, carotenoids etc., from fruit and vegetables) has been tackled in antioxidant supplementation experiments. The use of poorly validated assays for base oxidation has not helped us to reach a definitive answer; it seems that, in any case, the level of oxidative damage has been greatly exaggerated. DNA damaging agents lead to characteristic kinds of base changes (transitions, transversions, deletions). The investigation of the spectrum of mutations in cancer-related genes studied in tumour tissue should lead to a better understanding of the agents ultimately responsible for inducing the tumour. Similarly, studying mutations in a neutral marker gene (not involved in tumorigenesis) can tell us about the origins of the 'background' level of mutations. So far, interpretation of the growing databases is largely speculative. (ABSTRACT PMID- 10385878 TI - Environmental effects and exposures to manganese from use of methylcyclopentadienyl manganese tricarbonyl (MMT) in gasoline. AB - Methylcyclopentadienyl Manganese Tricarbonyl (MMT) has been used since the 1970s in the U.S. as a gasoline octane enhancer Extensive testing of the effects of MMT on regulated gaseous emissions carried out on a wide variety of automobiles showed that use of MMT resulted in significantly lower NOx emissions Tests showed that less than 15% of the manganese from MMT combustion was emitted from the tailpipe, mostly in the PM2.5 fraction as manganese phosphate, with some manganese sulfate and a very small amount of manganese oxide. MMT has been used in Canada in virtually all unleaded gasoline for about 20 years. A probability based study involving over 900 personal exposure samples in Toronto confirmed exposures to airborne PM2.5 Mn in the general population are quite low (.008 microgram/m3-median). Ambient levels of airborne manganese in Toronto are about the same as those in areas where MMT is not used. Exposures to manganese among the general population in Toronto are well within safe limits determined by the U.S. EPA and other standard setting bodies around the world. PMID- 10385879 TI - Airborne manganese particulates and methylcyclopentadienyl manganese tricarbonyl (MMT) at selected outdoor sites in Montreal. AB - This study aims to assess the atmospheric concentrations of methylcyclopentadienyl manganese tricarbonyl (MMT), respirable manganese (MNR) and total manganese (MnT) in certain specific microenvironments and to provide an estimation of human exposure to MnR. Sampling was carried out in five microenvironments: a gas station, an underground car park, downtown Montreal, near an expressway and near an oil refinery. The samples were collected using Gil Air portable pumps during three days and were analyzed by instrumental neutron activation analysis (INAA). The mean concentrations of MnR, MnT and MMT were 0.036 microgram m-3, 0.103 microgram m-3 and 0.005 microgram m-3 respectively. The MnR/MnT ratios vary from 25% to 43% (mean 35%) while the MMT/MnT ratios averaged about 5%. Furthermore, the mean concentration of the MnR measured near the expressway (0.053 microgram m-3) is similar to the United States Environmental Protection Agency (U.S. EPA) reference concentration (RfC = 0.05 microgram m-3). The average daily environmental exposure dose to MNR is estimated at 0.010 microgram kg-1 d-1 and its contribution to the multimedia exposure (air, food and water) is low. The overall results show a lack of potential exposure to MMT and substantial concentrations of MnR near an expressway. PMID- 10385880 TI - Pharmacokinetic data needs to support risk assessments for inhaled and ingested manganese. AB - Manganese (Mn)-deficiency or Mn-excess can lead to adverse biological consequences. Central nervous system tissues, rich in dopaminergic neurons, are the targets whether the Mn gains entrance by inhalation, oral ingestion, or intravenous administration. Risk assessments with Mn need to ensure that brain concentrations in the globus pallidus and striatum stay within the range of normal. This paper first provides a critical review of the biological factors that determine the disposition of Mn in tissues within the body. Secondly, it outlines specific data needs for developing a physiologically based pharmacokinetic (PBPK) model for Mn to assist in conducting risk assessments for inhaled and ingested Mn. Uptake of dietary Mn appears to be controlled by several dose-dependent processes: biliary excretion, intestinal absorption, and intestinal elimination. Mn absorbed in the divalent form from the gut via the portal blood is complexed with plasma proteins that are efficiently removed by the liver. Absorption of Mn via inhalation, intratracheal instillation or intravenous infusions bypasses the control processes in the gastrointestinal tract. After absorption into the blood system by these alternate routes, Mn is apparently oxidized by ceruloplasmin and the trivalent Mn binds to the iron carrying protein, transferrin. Brain uptake of Mn occurs via transferrin receptors located in various brain regions. Transferrin-bound trivalent Mn is not as readily removed by the liver, as are protein complexes with divalent Mn. Thus, Mn delivered by these other dose routes would be available for uptake into tissues for a longer period of time than the orally administered Mn, leading to quantitative differences in tissue uptake for different dose routes. Several important data gaps impede organizing these various physiological factors into a multi-dose route PK model for Mn. They include knowledge of (1) oxidation rates of Mn in blood, (2) uptake rates of protein-bound forms of Mn by the liver, (3) neuronal transfer rates within the CNS, and (4) quantitative analyses of the control processes that regulate uptake of ingested Mn by the intestines and liver. These data gaps are the main obstacles to developing a risk assessment strategy for Mn that considers contributions of both inhalation and ingestion of this essential nutrient in determining brain Mn concentrations. PMID- 10385882 TI - Uptake of metals in the brain via olfactory pathways. AB - In the olfactory epithelium the dendrites of the primary olfactory neurons are in contact with the nasal lumen, and via the axons these neurons are also connected to the olfactory bulbs of the brain. Materials which come into contact with the olfactory epithelium can be taken up in the primary olfactory neurons and be transported to the olfactory bulbs and even further into other areas of the brain. The present review deals with the mechanism of uptake and transport of metals in the olfactory system. Metals discussed are mainly manganese, cadmium, nickel and mercury. Among the metals so far examined, manganese has been found to have a unique capacity to be taken up via the olfactory pathways and pass transneuronally to other parts of the brain. It is considered that the occupational neurotoxicity of inhaled manganese may be related to an uptake of the metal into the brain via the olfactory pathways. Studies with nickel indicate that this metal, following a transport to the terminal parts of the primary olfactory neurons in the glomeruli of the bulbs, slowly passes to secondary and tertiary olfactory neurons. Cadmium and mercury are transported along the primary olfactory neurons to their terminations in the olfactory bulbs, but these metals appear unable to continue along secondary olfactory neurons. Occupational inhalation of nickel or cadmium can be toxic to the olfactory sense. It is not yet known whether mercury is toxic to the olfactory system in mammals, but this metal is known to alter olfaction and olfactory-related behaviour in fish. Data in the literature dealing with a potential olfactory-related neurotoxicity of aluminum are also discussed in the paper. PMID- 10385881 TI - Manganese uptake and distribution in the central nervous system (CNS). AB - Information about the nature of manganese (Mn)-binding ligands in plasma and serum, and its transport mechanism across the blood-brain barrier (BBB) is sparse. Most studies to date have focused on distribution, excretion, and accumulation of intravenous and intraperitoneal solutions of soluble divalent salts of Mn. Mn is transported in the blood primarily in the divalent oxidation state (Mn2+) and crosses the BBB via specific carriers at a rate far slower than in other tissues. Mn transport across the BBB occurs both in the 2+ and 3+ oxidation state. Within the CNS, Mn accumulates primarily within astrocytes, presumably because the astrocyte-specific enzyme, glutamine synthetase (GS), represents an important regulatory target of Mn. Compared to Mn2+, Mn3+ has a slower elimination rate and therefore, may have a greater tendency to accumulate in tissues. Furthermore, in view of the dependence of Mn accumulation within the CNS on iron (Fe) homeostasis, the oxidation state of Mn may represent a key determinant in the differential distribution, accumulation and secretion profiles of Mn, a fact that has received little attention in experimental biology toxicology. Accordingly, the distribution and membrane transport of Mn emphasizes the importance of: 1) the oxidation state of Mn, as it governs the affinity of Mn to endogenous ligands, and 2) the reaction of Mn3+ with transferrin, the plasma iron-carrying protein. This review will focus on transport kinetics of Mn across the BBB (both in the 2+ and 3+ oxidation state), the putative role of transferrin in the transport of Mn across the BBB, the transport of Mn by astrocytes, as well as the physiological significance of Mn to the function GS. PMID- 10385883 TI - Distribution of manganese in development. AB - Elimination of manganese is closely related to uptake in the normal adult and is believed to play a critical role in maintaining manganese homeostasis in the face of changing manganese intake. Data from immature rats, mice and cats have suggested that elimination of manganese undergoes a period of maturation with adult patterns of excretion developing at about the time of weaning. In addition, the uptake of manganese from the intestine appears to be more efficient in young animals than in adults. These two sets of findings raise the possibility that exposure to elevated manganese levels during the perinatal period might yield excessive concentrations of this metal in the developing organism. Such an outcome might lead to manganese accumulations in organ systems where subsequent mobilization might be difficult and might produce permanent toxic injury. This review evaluates the patterns of manganese uptake and distribution following prenatal and pre-weaning exposure using a variety of model systems. The data demonstrate that manganese does cross the placenta and enter fetal tissue although the extent of material crossing the placenta appears to be limited. The issue of neonatal manganese elimination following tracer and toxic exposure levels to manganese is addressed. The data show that that the neonatal rodent is significantly more effective in eliminating manganese than previously believed based upon tracer studies. Finally, data are presented on regional brain manganese distribution. These data highlight the lack of agreement on whether manganese is concentrated in specific brain areas. PMID- 10385884 TI - Nutrition versus toxicology of manganese in humans: evaluation of potential biomarkers. AB - Manganese intake can vary greatly with food choices, water composition, and supplement use. Thus, individuals consuming Western diets consume from < 1 to > 10 mg Mn/d. The levels of manganese intake associated with adverse effects (both deficient and toxic) are debatable. Moreover, many of the symptoms of manganese deficiency (growth retardation, changes in circulating HDL cholesterol and glucose levels, reproductive failure) and manganese toxicity (growth depression, anemia) are non-specific. The bone deformities observed in manganese-deficient animals and neurological symptoms of individuals who have inhaled excess manganese are permanent and illustrate the need to identify sensitive biomarkers of manganese status that appear before these symptoms. Manganese balance and excretion data are not useful biomarkers of manganese exposure but demonstrate that the body is protected against manganese toxicity primarily by low absorption and/or rapid presystemic elimination of manganese by the liver. Serum manganese concentrations in combination with lymphocyte manganese-dependent superoxide dismutase (MnSOD) activity and perhaps blood arginase activity, appear to be the best ways to monitor ingestion of insufficient manganese. Serum manganese concentrations in combination with brain MRI (magnetic resonance imaging) scans, and perhaps a battery of neurofunctional tests, appear to be the best ways to monitor excessive exposure to manganese. PMID- 10385885 TI - Nutritional aspects of manganese from experimental studies. AB - In experimental animals, dietary manganese deficiency can result in numerous biochemical and structural abnormalities. Deficient animals can be characterized by impaired insulin production, alterations in lipoprotein metabolism, an impaired oxidant defense system, and perturbations in growth factor metabolism. If the deficiency occurs during early development, there can be pronounced skeletal abnormalities and an irreversible ataxia. Several lines of evidence suggest that manganese deficiency may be a problem in some human populations. Manganese toxicity can also pose a significant health risk. In experimental animals, acute manganese toxicity can result in numerous biochemical pathologies. However, the above occurs typically when the manganese is given via injection; most animals show considerable resistance to dietary manganese toxicosis. Similarly, confirmed cases of manganese toxicity in humans are currently restricted to cases of exposure to high levels of airborne manganese, and to cases when manganese excretory pathways are compromised. PMID- 10385886 TI - Manganese neurotoxicity: a review of clinical features, imaging and pathology. AB - Manganese intoxication can result in a syndrome of parkinsonism and dystonia. If these extrapyramidal findings are present, they are likely to be irreversible and even progress after termination of the exposure to manganese. Clinical features are usually sufficient to distinguish these patients from those with Parkinson's disease. The neurological syndrome does not respond to levodopa. Imaging of the brain may reveal MRI signal changes in the globus pallidus, striatum, and midbrain. Positron emission tomography reveals normal presynaptic and postsynaptic nigrostriatal dopaminergic function. The primary site of neurological damage has been shown by pathological studies to be the globus pallidus. The mechanism of toxicity is not clear. PMID- 10385888 TI - Idiopathic parkinsonism with superimposed manganese exposure: utility of positron emission tomography. AB - It is difficult to distinguish manganism from idiopathic parkinsonism by clinical signs only. Case history and examination: A 48-year-old welder for over 10 years complained of masked face, right side (arm and leg) resting tremor, and bradykinesia for over one year. Magnetic resonance imaging (MRI) findings showed symmetrical high signal intensities in the globus pallidus on T1 weighted image. These intensities disappeared almost completely six months after cessation of exposure. 18F-6-fluorodopa (18F-dopa) positron emission tomography (PET) findings showed reduced 18F-dopa uptake in the left putamen, findings which appear in idiopathic parkinsonism. A PET study is necessary to distinguish manganism from idiopathic parkinsonism, especially in a working environment with elevated Mn concentrations, such as welding. PMID- 10385887 TI - Occupational exposure to manganese, copper, lead, iron, mercury and zinc and the risk of Parkinson's disease. AB - A population-based case-control study was conducted in the Henry Ford Health System (HFHS) in metropolitan Detroit to assess occupational exposures to manganese, copper, lead, iron, mercury and zinc as risk factors for Parkinson's disease (PD). Non-demented men and women 50 years of age who were receiving primary medical care at HFHS were recruited, and concurrently enrolled cases (n = 144) and controls (n = 464) were frequency-matched for sex, race and age (+/- 5 years). A risk factor questionnaire, administered by trained interviewers, inquired about every job held by each subject for 6 months from age 18 onward, including a detailed assessment of actual job tasks, tools and environment. An experienced industrial hygienist, blinded to subjects' case-control status, used these data to rate every job as exposed or not exposed to one or more of the metals of interest. Adjusting for sex, race, age and smoking status, 20 years of occupational exposure to any metal was not associated with PD. However, more than 20 years exposure to manganese (Odds Ratio [OR] = 10.61, 95% Confidence Interval [CI] = 1.06, 105.83) or copper (OR = 2.49, 95% CI = 1.06,5.89) was associated with PD. Occupational exposure for > 20 years to combinations of lead-copper (OR = 5.24, 95% CI = 1.59, 17.21), lead-iron (OR = 2.83, 95% CI = 1.07,7.50), and iron-copper (OR = 3.69, 95% CI = 1.40,9.71) was also associated with the disease. No association of occupational exposure to iron, mercury or zinc with PD was found. A lack of statistical power precluded analyses of metal combinations for those with a low prevalence of exposure (i.e., manganese, mercury and zinc). Our findings suggest that chronic occupational exposure to manganese or copper, individually, or to dual combinations of lead, iron and copper, is associated with PD. PMID- 10385889 TI - Prospective study on the reversibility of neurobehavioral effects in workers exposed to manganese dioxide. AB - In 1987, a cross-sectional study in a dry-alkaline battery plant in Belgium revealed subclinical neurobehavioral dysfunctions associated with inhalation exposure to manganese dioxide (MnO2) particulate. The overall geometric mean of the time-weighted average concentration of manganese (Mn) in "total" dust (MnT) amounted, at that time, to 1 mg Mn/m3 and the duration of exposure was 5.5 years on average. An 8-year longitudinal investigation was conducted in this cohort (n = 92) in order to find out whether early effects on eye-hand coordination (EHC), hand steadiness (HST), and simple visual reaction time (VRT) were reversible when the airborne manganese concentration at the workplace was abated. During the observation period from 1988 to 1995, MnT monitoring was implemented on a monthly basis producing more than 1300 personal air samples, EHC tests were given yearly to assess the precision of the hand-forearm movement (PN1), and HST and VRT tests were carried out yearly since 1991. By the end of the study, the cohort size had dropped to 34 subjects. The model of unbalanced repeated measurements with unstructured covariance matrix and a time-varying covariate (log MnT) was the most appropriate to analyze the data. Wald chi 2 statistic was used for testing time-trends. The reduction of MnT over time was significantly associated with an improvement of the PN1 values (total cohort: Wald chi 2 = 8.5, p = 0.004; beta log MnT = -6.098 +/- 2.096). Like in the total cohort, time-trends were also found in the three exposure subgroups which could be identified in the cohort (average MnT over 1987-1992 were about 400, 600, and 2000 micrograms Mn/m3 for the low, medium, and high exposure subgroups, respectively). Only in the low exposure subgroup the PN1 value normalized when MnT (provisional estimates) decreased from about 400 to 130 micrograms Mn/m3 by the end of the study. Solely the reduction in MnT explained these findings on PN1, while a "healthy-worker effect" mechanism was unlikely to have operated. The prognosis for the medium and high exposure subgroups remains uncertain as the improvement of their EHC performance may have been affected by past MnO2 exposure to such an extent that the persistence of a partial loss of EHC ability is suggested. The time courses of the HST and VRT test results, however, indicated the absence of any improvement, suggesting irreversible impairment of hand stability (postural tremor) and simple visual reaction time. A separate examination in a group of 39 control subjects, re-tested 10 years after the first test in 1987, virtually precluded age as confounding factor in this prospective study. The findings of the longitudinal study are corroborated by the outcome of a separate follow-up study in a group of 24 ex-Mn employees, who showed in 1996 a significant improvement of eye-hand coordination after at least three years with no MnO2 exposure; as to HST and VRT, there was no significant change in the deficit of these two neurobehavioral markers. PMID- 10385890 TI - Results from eleven years of neurological health surveillance at a manganese oxide and salt producing plant. AB - In 1983, Roels et al. (1987a, b) collected blood and urine samples and conducted neurological testing of workers at a manganese oxide and salt producing plant in Belgium, and at a nearby chemical plant. Workers from the manganese plant performed significantly worse than workers from the chemical plant on tests of short-term memory capacity, eye-hand coordination, hand steadiness, and visual reaction time. Between 1985 and 1996, workers at the manganese plant were tested routinely using the same battery of neurological tests and biological sampling that were employed by Roels et al. Blood and urine Mn levels remained comparable throughout the eleven years of testing to those measured in 1983 by Roels et al. On a gross basis, neurological test results during this period were comparable or superior to results obtained by Roels et al., despite the fact that workers were older and had been exposed longer. Large year-to-year differences were observed in some neurological test outcomes that could not be explained by age or Mn exposure. Older age was significantly associated with poorer performance on tests of short-term memory and eye-hand coordination. After controlling for age and year of testing, reduced hand steadiness was significantly associated with blood Mn and (Marginally) urine Mn, and both reaction time and one measure of hand steadiness were significantly associated with years of Mn exposure. No significant associations were found between any measure of Mn exposure and results from either short-term memory or eye-hand coordination tests. Investigations regarding whether neurological scores of individual workers studied by Roels et al. continued to worsen with continued occupational Mn exposure were hampered by lack of a suitable comparison group. However, there was no evidence that neurological effects seen earlier in these workers by Roels et al. were progressing towards clinical detectable signs. PMID- 10385891 TI - Long-term exposure to "low levels" of manganese oxides and neurofunctional changes in ferroalloy workers. AB - Occupational exposure to manganese can cause early neurobehavioral effects in low or a-symptomatic workers. A battery of neuropsychological tests was administered to a group of 61 ferroalloy male workers and 87 controls. The average (geometric mean) manganese concentrations in total dust at the plant have changed from 1981 to 1997 respectively from 1597.03 micrograms/m3 to 239 micrograms/m3 at the furnace area; from 151.53 to 255.76 micrograms/m3 at the casting area; from 167 to 54.7 micrograms/m3 at the maintenance (welding operations), yielding a current overall value of 54.25 micrograms/m3. A cumulative exposure index was calculated for each alloy worker and the average value (geometric mean) resulted to be 1204.87 micrograms/m3 x years, which divided by the average length of exposure (15.17 years), showed the concentration of 70.83 micrograms/m3 of manganese in total dust. Blood and urinary manganese geometric means resulted significantly higher in the exposed workers (9.18 micrograms/l and 1.53 micrograms/g creatinine, respectively) than in controls (5.74 micrograms/l and 0.40 microgram/g creatinine, respectively). A positive correlation was observed between the airborne manganese concentrations in total dust and blood manganese (n = 55; R = 0.36; R2 = 0.13; p = 0.0068), whereas no association resulted between cumulative exposure index and both blood manganese and urinary manganese. Higher prevalence of symptoms reporting was observed in the alloy workers concerning irritability, loss of equilibrium and rigidity. Tremor parameters including the central frequency and its dispersion, resulted to be statistically different in the exposed workers compared to the controls. Motor functions exploring the coordination of rapid and alternating movements and memory functions resulted to be impaired in the manganese workers. Dose-effect relationships were observed between the cumulative exposure index and some of the test results, whereas no relationship was found with the airborne manganese concentrations and the biological indicators of exposure. These findings are consistent with the existing knowledge of a cumulative mechanism of action of manganese, which must be carefully considered when setting safe exposure levels. In order to be protective for the entire working life, the average annual exposure level should be lower than 100 micrograms/m3. PMID- 10385892 TI - Focused medical surveillance: a search for subclinical movement disorders in a cohort of U.S. workers exposed to low levels of manganese dust. AB - Seventy-five workers with recent and/or historical exposure to manganese (Mn) at a metal producing plant in northern Mississippi were closely matched with 75 control workers who had no known history of occupational exposure to Mn. Both plants are OSHA STAR work sites and share common medical, safety, and industrial hygiene services. Airborne Mn levels were assessed for each of twelve job categories at the Mn facility by collecting 63 side-by-side full-shift personal samples of both total and respirable Mn dust. Exposures of workers currently working with Mn averaged 0.066 mg/3 respirable and 0.18 mg/3 total Mn. An assessment of major equipment and work practice changes over the past several years and estimates of the resultant relative impacts on exposure was made. Based on this information and individual employment information, each worker's cumulative exposure to respirable and total Mn was estimated for the preceding 30 days, preceding year, and for the worker's entire employment history. Both Mn and control workers were administered multiple neuropsychological tests including tests of hand-eye coordination, hand steadiness, complex reaction time, and rapidity of finger tapping. A questionnaire was used to evaluate a worker's neuropsychological status. Performance decreased significantly with increasing age in tests of hand-eye coordination, complex reaction time and finger tapping speed. No effect of Mn exposure was found on the results of the questionnaire or any neuropsychological test. PMID- 10385893 TI - Manganese neurotoxicity in industrial exposures: proof of effects, critical exposure level, and sensitive tests. AB - Manganese neurotoxicity has been known for more than 150 years, since Couper (1837) described a syndrome, similar to Parkinson's disease, in Scottish workers exposed to high levels of dust while grinding "black oxide of manganese" at a chemical industry. Since then, the syndrome has been described in several groups of highly exposed miners and other workers. A thorough review of manganese neurotoxicity was provided by the WHO (1981) and a recent update was written by Mergler and Baldwin (1997). From these reviews it is evident that the critical effect from manganese exposure is damage to the central nervous system, and that the effects, once established, are generally irreversible. Therefore, the early detection of symptoms of manganese neurotoxicity in populations at risk is of the utmost importance. In spite of this fact, only about a dozen studies of manganese exposed groups of workers have been performed using psychological test methods. These studies are briefly presented, the preponderance of proof for Mn neurotoxicity even in present industrial settings is demonstrated, the critical exposure level is briefly discussed, the test methods are evaluated, and recommendations for a test battery useful for studies of manganese neurotoxicity, are presented. PMID- 10385894 TI - Manganese neurotoxicity, a continuum of dysfunction: results from a community based study. AB - Excessive manganese (Mn) has been associated with neurobehavioral deficits and neurological and/or neuropsychiatric illness, but the level at which this metal can cause adverse neurotoxic effects, particularly with long-term exposure, is still unknown. The objective of the present study was to assess nervous system functions in residents exposed to manganese from a variety of environmental sources. A random stratified sampling procedure was used to select participants; persons with a history of workplace exposure to Mn and other neurotoxic substances were excluded. A self-administered questionnaire provided data on socio-demographic variables. Blood samples were analyzed for total manganese (MnB), lead, mercury and serum iron. Nervous system assessment included computer and hand-administered neurobehavioral tests, computerized neuromotor tests, sensory evaluation and a neurological examination. The present analyses include 273 persons (151 women and 122 men); MnB range: 2.5 micrograms/L-15.9 micrograms/L (median: 7.3 micrograms/L). Multivariate analyses were used and neuro-outcomes were examined with respect to MnB, taking into account potential confounders and covariables. Results were grouped according to neurofunctional areas and MANOVA analyses revealed that higher MnB (7.5 micrograms/L) was significantly associated with changes in coordinated upper limb movements (Wilks' lambda = 0.92; p = 0.04) and poorer learning and recall (men: Wilks' lambda = 0.77; p = 0.002; women: Wilks' lambda = 0.86; p = 0.04). Further analyses revealed that with increasing log MnB (Simple regression: p < 0.05) performance on a pointing task was poorer, frequency dispersion of hand-arm tremor decreased, while harmonic index increased, and the velocity of a pronation/supination arm movement was slower. An Mn-age interaction was observed for certain motor tasks, with the poorest performance observed among those _50 y and in the higher MnB category. Differences between genders suggest that men may be at greater risk than women, although effects were also observed in women. These findings are consistent with the hypothesis that Mn neurotoxicity can be viewed on a continuum of dysfunction, with early, subtle changes at lower exposure levels. PMID- 10385895 TI - Bioindicator and exposure data for a population based study of manganese. AB - Exposure data and bioindicators were obtained for a study whose objective was detection of early manifestations of manganese (Mn) neurotoxicity in a population with potential environmental exposure. The study included persons with no history of neurotoxic workplace exposure in Southwest Quebec, drawn from seven postal code regions, defining a set of geographically contiguous zones. Blood samples were analyzed for total Mn (MnB), lead (PbB), total mercury (HgT) and serum iron (FeS). Drinking water samples from participants' residences were analyzed for manganese (MnW). At 4 sites, limited 24-hour high volume air samples for total particulates (TP) and PM10, were analyzed for Mn and Pb. Sociodemographic and dietary information was obtained by self-administered questionnaire. The geometric mean (GM) for MnB values (n = 297) was 7.14 micrograms/L. Levels of MnB in women (n = 156; GM 7.50 micrograms/L) were significantly higher than in men (n = 141; GM 6.75 micrograms/L). No relationship was found between MnB and PbB or HgT. FeS was significantly higher in men (GM 18.38 mumol/L) than women (GM 15.0 mumol/L). For women, MnB was correlated to FeS, with a tendency to decrease with increasing age. For men, no relationship was found between MnB levels and either FeS or age, although FeS showed a strong inverse relationship with age. The 24 hour mean levels of MnTP at the 4 sites varied between 0.009 microgram/m3 and 0.035 microgram/m3; intersite differences were not significant. For Mn in PM10 (MnPM10), mean values ranged from 0.007 microgram/m3 to 0.019 microgram/m3; intersite differences were significant. A total of 278 MnW samples were obtained, 16 from residences served by wells. The GM for MnW was 4.11 micrograms/L (range: 0.50-71.1 micrograms/L, excluding wells; MnW for wells ranged from non-detectable to 158.9 micrograms/L. Individually, there was no relation between MnW and MnB. Geographic analysis of the MnB and MnW data by an algorithm grouping contiguous postal code zones, combined with air data, lead to definition of a geographic parameter, distinguishing two regions relative to a former manganese alloy plant, which contributed significantly to MnB. A multiple regression model was developed, explaining 6.7% of the variability in MnB (F = 5.12; p < 0.001); when controlling for gender, geographic region with higher levels of airborne manganese and the frequency of consumption of cereals and leaf vegetables contributed positively to MnB levels, while serum iron was negatively related. PMID- 10385896 TI - Quantification of neuromotor function for detection of the effects of manganese. AB - The effect of low level exposure to manganese (Mn) was examined in 297 subjects from southwest Quebec. Blood manganese (MnB) levels as well as other possibly relevant variables were obtained. We tested equipment and analysis procedures that we have developed to quantify aspects of motor function thought to be affected by exposure to toxins, in particular, rapid alternating movements, rapid and precise pointing movements, and tremor. (1) The eurythmokinesimeter measures timing and precision of contacts between a hand-held stylus and a pair of metal targets (proximal/distal). This roughly approximates the finger-to-nose test of the UPDRS. Characteristics quantifying speed, precision and regularity of the movements were calculated, as well as multiple contacts due to tremor and an index based on Fitts' Law eliminating the effect of the trade-off between speed and precision. (2) The diadochokinesimeter accurately measures rapid rotation of the forearms (pronation/supination). Characteristics quantifying the range, speed, period, shape and regularity of the oscillatory movements were calculated, as well as the smoothness of the movement on a fine scale and the coordination between the two hands. (3) Postural tremor of the arm and hand was measured using the accelerometry-based "TREMOR" system of Danish Product Development. We used the amplitude and frequency characteristics provided by the TREMOR system: intensity, center frequency, dispersion and harmonic index. Previous studies have shown that these tests are sufficiently sensitive to detect small differences in performance of different groups of subjects, with indications that some characteristics are also specific to particular conditions. In this study, significant though small effects related to age and gender were found in many of the characteristics. When effects of other variables are removed, low-level exposure to Mn was found to be associated with a decrease in ability to perform regular, rapid and precise pointing movements, a decrease in ability to attain high maximum rotation speeds in rapid alternating movements, and an increase in regularity of tremor oscillations. Moreover, the effects are age-related for levels of MnB 7.5, micrograms/L. PMID- 10385897 TI - Neuropsychiatric effects of manganese on mood. AB - Adverse mood effects of overexposure to Manganese (Mn) have been described in 15 studies which frequently report an association of Mn exposure with adverse effects in six dimensions of mood: 1) anxiety, nervousness, irritability; 2) psychotic experiences; 3) emotional disturbance; 4) fatigue, lack of vigor, sleep disturbance; 5) impulsive/compulsive behavior; 6) aggression hostility. Only 1:15 studies used a standardized psychological measure of mood, while the current study of environmental Mn exposure used two standardized mood scales in evaluating low levels of Mn exposure and mood sequelae. The Profile of Moods State (POMS) and Brief Symptom Inventory (BSI) were used, and results indicate that men who are older and have higher Mn levels show significant disturbances on four of the six mood dimensions. Increased scores were seen in the anxiety, nervousness, irritability; emotional disturbance; and aggression, hostility dimensions relative to those who had lower levels of Mn. The BSI and POMS are useful adjuncts in the assessment of mood/Mn effects. PMID- 10385898 TI - Effects from environmental Mn exposures: a review of the evidence from non occupational exposure studies. AB - OBJECTIVE: The risk posed to human health by environmental manganese exposure is unknown. Occupational-exposure outcomes may not extrapolate to environmental exposures due to the healthy worker effect and differences in dosage parameters which may affect the biological response. This paper attempts to combine the existing literature on non-occupational Mn exposures with results from our current study in SW Quebec on environmental Mn exposure (Mergler et al., this issue) within the framework of a biologically-based, dose-response (BBDR) model. BBDR MODEL: The basic BBDR model consists of seven stages relating exposure to health effects. The stages are: 1) sources, 2) applied dose, 3) absorbed dose, 4) target-site dose, 5) toxic event, 6) measurable change, and 7) health outcome. RESULTS: Several air monitoring programs, such as the PTEAM study (Riverside, CA, 1990, mean PM10 Mn outdoor-airborne 24 h average = 0.045 microgram/m3), provided data relevant to the estimation of Mn applied dose, but did not include measures of body burden. Data from the SW Quebec study showed a mean total-particulate airborne Mn concentration of 0.022 microgram/m3 with a range of 0.009 to 0.035 microgram/m3 across four sampling sites, whereas the EPA reference concentration (RfC) is 0.05 microgram/m3. EPA has considered tap water levels to be safe below 200 micrograms/l Mn, and mean Mn tap-water (MnW) level in the participants' homes was 6.38 +/- 11.95 micrograms/l with a range from 0.1 to 158.9 micrograms/l Mn. A previous study of MnW exposure in Greece reported Mn levels in areas with low, medium and high MnW ranging from 4 to 2,300 micrograms/l and a significant association with Mn in hair but not Mn in blood (MnB). The mean absorbed dose of the SW Quebec study participants, as indicated by MnB, was 7.5 +/- 2.3 micrograms/l with a range of 2.5 to 15.9 micrograms/l. Our study and others on environmental Mn exposure did not provide an estimate of target-site dose. However, a significant correlation (r = 0.65) between MnB and signal intensity in T1-weighted MRI images has been reported in liver-disease patients with Parkinson like signs who had MnB levels as low as 6.6 micrograms/l. Only animal and in vitro studies have provided evidence on the mechanisms of toxicity caused by Mn in the CNS. Several studies reported measurable changes in endpoints suggestive of a Parkinson-like syndrome in subjects with MnB levels ranging from 7.5 to 25.0 micrograms/l. Among other effects on neurobehavioral function observed in the current study was a significant relationship between MnB and the direction and speed of body-sway in men. The effects observed in these participants are sub clinical and no health outcomes have been diagnosed. However, the Parkinson's disease incidence in the study area was previously reported to be 2-5 times higher than in the rest of Quebec, and several studies indicate that 25-35% of idiopathic Parkinson disease diagnoses are incorrect. Our study, the Greek study, and some clinical studies suggest that the risk of a Parkinson-like syndrome diagnosis may increase with continued Mn exposure and aging. CONCLUSION: The limited data available for the BBDR model point to the need for evidence, particularly on relationships between Mn species, exposure route, MnB with chronic environmental exposure, ageing, and susceptibility factors, to improve human-health risk assessments for chronic, environmental Mn exposure. PMID- 10385899 TI - Peripheral biomarkers and exposure to manganese. AB - Biochemical mechanisms underlying manganese (Mn) toxicity include dopamine (DA) auto-oxidation and free radical generation with subsequent neuronal damage. A neuroendocrine approach based on the measurement of serum prolactin (PRL) has been proposed to assess the tonic inhibition of pituitary lactotrope cells by the tubero-infundibular DA system. Low level exposure to Mn oxides in industrial settings is associated with a shift in the distribution of serum PRL towards higher levels as compared to matched controls. The follow-up of a small cohort of workers from a ferro-manganese plant showed that the increased prevalence of abnormally high PRL values is stable over time. Although the mechanistic basis for their application is less straightforward, other biochemical markers such as dopamine beta hydroxylase and monoamine oxidase Type B, have also been assessed. Contrary to PRL levels, these markers cannot be recommended to monitor early biochemical effects of manganese exposure at the workplace. Early biochemical events can be modified by genetically determined individual differences. Owing to the possible role of a reduced capacity of glutathione conjugation as a risk factor increasing the susceptibility to the action of free radicals generated in the presence of Mn, the class mu glutathione S transferase (GSTM1) genotype has also been assessed in workers occupationally exposed. However, the GSTM1 null genotype does not appear to play an important role in the susceptibility to biochemical effects of Mn. A logistic model of the dose-response relationship based on urinary Mn as marker of exposure indicates that the benchmark dose corresponds to Mn levels as low as 0.4 microgram/l. This would imply that environmental exposure to Mn may contribute to abnormally high serum PRL in the general population. PMID- 10385900 TI - The feasibility of measuring manganese concentrations in human liver using neutron activation analysis. AB - Manganese is an element which is required by the human body. However, as with most metals, in large amounts manganese can be toxic. People who suffer from severe manganese intoxication have symptoms similar to those of Parkinson's disease. Preclinical symptoms of manganese intoxication have recently been detected in individuals working in industries which have manganese dioxide dust in the air. The concentration of many toxic elements can be measured in vivo using neutron activation. A small dose of neutrons is delivered to the organ of interest, the neutrons are readily captured by the target nuclei, and the gamma rays given off can be detected outside of the body. A neutron activation analysis system is being developed to measure manganese concentrations in humans. The McMaster KN-accelerator supplies the neutron beam and the thermal neutron capture reaction 55Mn(n,gamma)56Mn is used. The half-life of 56Mn is 2.58 hr and thus counting can occur after irradiation. The 847 keV gamma ray given off when 56Mn decays is detected using a Nal detector. Calibration curves are made using phantoms with known concentrations of Mn. This system will be used to monitor manganese levels in individuals who have occupational exposure to the element. Preliminary measurements, using liver phantoms, give a minimum detectable limit for Mn in the liver of less than one part per million, which is well below normal levels. PMID- 10385901 TI - Animal models of manganese's neurotoxicity. AB - Manganese's neurotoxicity continues to present a puzzling array of differences across individuals and across published reports in the profile of effects seen in humans and nonhuman species, but some of the sources of individual variability are becoming clear from studies of animals. The kinetics of manganese is a critical component of any assessment of risk associated with exposure. After inhalation, the uptake of manganese into and elimination from the central nervous system are slow and some manganese remains in the nervous system a year after inhalation. Comparison with other parenteral routes suggests that manganese depots in lung prolongs exposure even after environmental exposure has ended. Manganese's neurotoxicity is associated with its appearance in basal ganglia structures, especially the globus pallidus. Manganese also appears in the pituitary gland but the functional consequences of this are not well understood. Other critical components in characterizing manganese's neurotoxicity appear to be the behavioral endpoints used, the species studied, and the exposure rate. Over neurological signs and excitability are associated with high exposure rates and the appearance of manganese throughout basal ganglia and basal forebrain regions. More focused behavioral endpoints are required to detect the subtle signs associated with slow exposure rates low exposure levels, but when such designs are used the effect is unequivocal. At lower exposure levels, doses of 5 mg/kg and greater, deficits in a task in which a monkey executed a rowing type motion against a spring approximating its body weight were clearly related to manganese exposure while other traditional measures of response patterns under schedules of reinforcement remained intact. Excitability and other signs of emotionality have not been reported at low exposure rates. In rodents, manganese accumulation and alterations in the function or concentration of neurotransmitters have been reported. Investigations of behavioral effects in these species, which usually involved locomotor activity, have resulted in less consistent results. Manganese produces a constellation of neurotoxic signs whose appearance and detection are influenced by dose and exposure rate. Despite investigations of manganese's neurotoxicity in animals over a wide range of exposure levels, a NOAEL has not been identified. PMID- 10385902 TI - Manganese mineral interactions in brain. AB - Manganese (Mn) is an essential mineral but is toxic when taken in excess. However, whether its interactions with other minerals in organs and cells are involved in mechanisms underlying Mn toxicity is poorly understood. We designed a developmental rat model of chronic Mn treatment (Group A: 1 mg MnCl2.4H2O per ml of drinking water; Group B: 10 mg MnCl2.4H2O per ml of drinking water; Group C: 20 mg MnCl2.4H2O per ml of drinking water; Control Group given water without manganese addition). Employing the model and instrumental neutron activation analysis, we investigated two hypotheses: (i) chronic manganese treatment alters the brain regional distribution of manganese and this altered manganese distribution also leads to region-specific changes of other metals; (ii) chronic manganese treatment induces differential changes in subcellular distributions of metals and electrolytes. In the treated rats, brain Mn level showed dose-related increases, the most pronounced being noted in striatum, hypothalamus, and hippocampus: these increases also led to alterations in regional distribution pattern of Mn. In the treated rats, Fe level was increased in hypothalamus, cerebellum, hippocampus, pons and medulla, and striatum. Cu level was increased in pons and medulla, hippocampus, midbrain, and striatum. Se level was increased in cerebellum, striatum, midbrain, hypothalamus, and pons and medulla. Zn level was increased in hypothalamus and striatum. Ca level was increased in midbrain but decreased in cerebellum; however, Mg and Al levels were not markedly affected. In brains of Mn-treated rats, Mn levels in subcellular fractions were all increased, being especially marked in nuclei, mitochondria, and synaptosomes; the subcellular distributions of Fe, Cu, Zn, and Mg were differentially altered although those of Al and Ca were minimally affected. These results are consistent with our hypotheses and may have implications in manganese neurotoxicity. The cellular and molecular mechanisms underlying manganese-mineral interactions in brain are still poorly defined and merit further investigation. PMID- 10385903 TI - Manganese and calcium transport in mitochondria: implications for manganese toxicity. AB - Mn2+ is sequestered by liver and brain mitochondria via the mitochondrial Ca2+ uniporter. The mitochondrial Ca2+ uniporter is a cooperative transport mechanism possessing an external activation site and a transport site. Ca2+ binding to the activation site greatly increases the velocity of uptake of both Ca2+ and Mn2+. Electron paramagnetic resonance (EPR) shows that over 97% of the Mn2+ in the mitochondrial matrix is normally bound to the membrane or to matrix proteins. EPR measurements of manganese within living isolated mitochondria can be repeated for hours, and during this time most of the manganese remains in the Mn2+ state. Mn2+ is transported out of mitochondria via the very slow Na(+)-independent efflux mechanism, which is an active (energy-requiring) mechanism. Mn2+ is not significantly transported over the Na(+)-dependent efflux mechanism, which is the dominant efflux mechanism in heart and brain mitochondria. Mn2+ inhibits the efflux of Ca2+ through both of these efflux mechanisms, having an apparent Ki of 7.9 nmol/mg protein on the Na(+)-independent efflux mechanism and an apparent Ki of 5.1 nmol/mg on the Na(+)-dependent efflux mechanism. Mn2+ inhibition of Ca2+ efflux may increase the probability of the mitochondria undergoing the mitochondrial permeability transition (MPT). Intramitochondrial Mn2+ also inhibits State 3 mitochondrial respiration using either succinate or malate plus glutamate as substrate. The data suggest that Mn2+ depletes cellular energy supplies by interfering with oxidative phosphorylation at the level of the F1ATPase and at much higher concentrations, at Complex I. Effects such as these could lead to apoptosis in active neurons. PMID- 10385904 TI - Implications for atypical antioxidative properties of manganese in iron-induced brain lipid peroxidation and copper-dependent low density lipoprotein conjugation. AB - Our group recently observed that manganese prevents oxidative brain injury in the iron-induced parkinsonian animal model. It has also been suggested that manganese retards while copper promotes the development of atherosclerosis. In this report, we provide further evidence to support a controversial notion that manganese is an atypical antioxidant. Among transition metals, Cu2+ and Fe2+ (0.1 to 125 microM), but not Mn2+, converted hydrogen peroxide to reactive hydroxyl radicals via the Fenton reaction at pH 7.4. Iron's pro-oxidative rate is relatively slow, but it is accelerated further by ascorbate (50 microM) in 37 degrees C Dulbecco's phosphate buffered saline. Moreover, Mn2+ (0-80 microM) concentration dependently retarded diene conjugation of human low density lipoproteins stimulated by 5 microM Cu2+. This new result is consistent with our recent finding that Mn2+ (0 to 20 microM) does not initiate brain lipid peroxidation while it inhibits iron induced peroxidation of polyunsaturated fatty acids. These unexpected manganese results are somewhat at odds with a prominent theory that manganese is a prooxidative transition metal. Furthermore, iron and copper induced free radical generation and lipid peroxidation are suppressed by lowering the incubation temperature; this suggests that hypothermia may decrease the oxidative stress and damage in vivo. In conclusion, normal dietary intake of manganese may protect cells and neurons from oxidant stress through the inhibition of propagation of lipid peroxidation caused by hydroxyl radicals generated by pro-oxidative transition metals such as iron and copper. Potential therapeutical uses of manganese, manganese SOD mimetics and hypothermia for protecting brain neurons and vascular endothelial cells against oxidative stress and damage have been successfully demonstrated in both animal models and clinical trials. PMID- 10385905 TI - Central nervous system toxicity of manganese. II: Cocaine or reserpine inhibit manganese concentration in the rat brain. AB - Manganese concentrates in the ventral mesencephalon of male Sprague-Dawley rats after intrathecal administration of MnCl2. We tested the hypothesis that Mn concentration in the central nervous system (CNS), particularly in the ventral mesencephalon, is decreased by inhibiting dopamine reuptake using cocaine or by decreasing dopamine concentrations using reserpine. The intrathecal administration of Mn (250 micrograms Mn/rat as MnCl2) caused the Mn concentration in the ventral mesencephalon to increase from 0.57 to 31.8 micrograms Mn/g. Cocaine administration (8.6 mg/kg i.p.) thirty minutes prior to MnCl2 decreased ventral mesencephalon Mn to 3.3 micrograms Mn/g. By giving reserpine (5 mg/kg i.p.) 24 hours prior to MnCl2 the ventral mesencephalon Mn concentration was decreased from 29.9 micrograms Mn/g to 3.7 micrograms Mn/g. Intrathecal MnCl2 decreased the dopamine concentration in the caudate putamen by 40% six hours after administration. Cocaine or reserpine decreased the Mn concentration in the ventral mesencephalon, occipital pole, frontal lobe and caudate putamen but did not change the Mn concentration in the cerebellum. The results indicate that the mechanism(s) by which Mn is concentrated in many brain regions can be inhibited by cocaine, a dopamine reuptake inhibitor, or by reserpine, a dopamine depleter, and suggest that the Mn concentration in the CNS is related to dopamine reuptake and/or concentration. PMID- 10385906 TI - Manganese-induced developmental neurotoxicity in the CD rat: is oxidative damage a mechanism of action? AB - Inhalation of high concentrations of manganese (Mn) is associated with an extrapyramidal motor disorder in humans. Oxidative damage, mediated by increased levels of Mn in dopaminergic brain regions and mitochondria, is a hypothesized mechanism of action for Mn-induced neuronal degeneration and loss. To test this proposed mechanism, developing CD rats, which may be at an increased risk for Mn induced neurotoxicity, were exposed orally to 0, 25, or 50 mg/kg/day of MnCl2 from postnatal day (PND) 1 to 49 Brain regional and mitochondrial Mn levels, brain regional reactive oxygen species (ROS) levels, and whole-brain nuclear and mitochondrial 8-OHdG levels were used to evaluate Mn-mediated oxidative damage. High-dose Mn exposure was associated with increased spontaneous motor activity on PND 21 and decreased body weights on PND 49. On PND 21, Mn concentrations were increased in brain regions and mitochondrial fractions in both low- and high-dose groups. ROS levels were elevated in cerebellum but not striatum. On PND 49, Mn concentrations in brain regions and mitochondrial fractions were increased only in the high-dose group. Mn exposure did not significantly alter 8-OHdG levels in either mitochondrial or nuclear DNA. Selective uptake of Mn by the striatum or mitochondrial fraction was not demonstrated at either time point. These data allow us to conclude that oral exposure to high levels of Mn in developing CD rats resulted in increased brain regional and mitochondrial Mn levels, increased motor activity, and decreased body weights but not in selective accumulation of Mn in the striatum or mitochondrial fraction of any brain region or elevations in striatal ROS or whole-brain 8-OHdG levels. These findings do not support the hypothesis that oxidative damage, as assessed by ROS and 8-OHdG levels, is a mechanism of action in Mn-induced developmental neurotoxicity in the CD rat. PMID- 10385908 TI - A brief history of the neurobehavioral toxicity of manganese: some unanswered questions. AB - It was observed by Couper in 1837 that manganese dust produces a neurological syndrome characterized by muscle weakness, tremor, bent posture, whispered speech and excess salivation. The similarity of these symptoms to those of Parkinson's disease were not recognized for many years. In addition to its Parkinson-like effects, manganese produces behavioral symptoms in humans including nervousness, hallucinations, memory loss, cognitive problems, bizarre behaviors and flight of ideas. Despite these signs and symptoms, there have been few systematic attempts to study the effects of manganese on behavior using animal models. The need to better understand the effects of manganese on behavior is becoming more important due to the potential of increased environmental exposure to manganese due to its use, or proposed use as a gasoline additive in a number of countries. However, there is debate as to which manganese compounds should receive priority for testing, what route of administration should be used in this testing, what dosing regimens should be used, what species are appropriate for behavioral testing, and what behavioral tests should be selected. Research to answer these questions is needed so that the behavioral effects of manganese can be described comprehensively and the mechanisms underlying these effects can be understood. PMID- 10385907 TI - Manganese neurotoxicity: a mechanistic hypothesis. AB - This review provides a summary of the presentations and abstracts presented at the 15th International Neurotoxicology Conference which may contribute to an understanding of the mechanism and pathogenesis of manganese (Mn2+) neurotoxicity. We propose that an understanding of the pathogenesis of Mn2+ neurotoxicity must incorporate data on (1) the factors controlling Mn2+ uptake and distribution within the CNS, (2) account for the apparent selectivity of dopaminergic neurons, (3) analyze the role of mitochondrial dysfunction and (4) provide data to support or refute the role of oxidative injury in the genesis of toxicity. We propose a multifactor hypothesis coupling Mn2+ uptake with coincident transport of aluminum and iron. Selectivity of dopaminergic neurons is dependent upon interactions of Mn2+ with dopamine transport and the role of Mn2+ as a pro-oxidative toxicant in conjunction with changes in iron concentration. Within the synaptic milieu, Mn(2+)-mitochondrial interaction will influence mitochondrial--Ca2+ transport kinetics leading to defective mitochondrial function, decreased oxidative phosphorylation, decreased ATP and accumulation of reactive oxygen species. Under the influence of excessive depolarization, energy failure will occur leading to secondary activation of an excitotoxic state. These conceptual ideas provide for mechanistic based hypotheses and testing and are likely to lead to rational therapeutic avenues directed against Mn2+ neurotoxicity. PMID- 10385909 TI - Inhalation health risks of manganese: an EPA perspective. AB - In 1994, the U.S. Environmental Protection Agency (EPA) denied a petition by Ethyl Corporation to allow the use of methylcyclopentadienyl manganese tricarbonyl (MMT) in unleaded gasoline, because of health concerns related to the inhalation of manganese (Mn) particulate emissions from combusted MMT. Although Ethyl successfully challenged EPA's denial of the petition on legal grounds, issues raised in EPA's health risk assessment have not been resolved to date. This paper summarizes features of the EPA health risk characterization, which included the use of various statistical techniques to derive several estimates of inhalation reference concentration (RfC) values for Mn as alternatives to the established value of 0.05 microgram Mn/m3. An exposure assessment projected distributions of personal exposure levels to particulate Mn if MMT were used in all unleaded gasoline. It was estimated that exposure levels of 5-10% of the modeled population might exceed a possible alternative RfC value of 0.1 microgram Mn/m3. However, due to data limitations, the risk characterization for Mn/MMT could raise only qualitative concerns about potential public health impacts and was unable to provide a quantitative estimate of risk. To improve the risk characterization, better information on Mn/MMT population exposures and health effects is needed. Much of this information is expected to be obtained under provisions of Section 211 of the Clean Air Act. Among the specific issues that remain to be resolved are the form or forms of Mn emitted from the combustion of MMT in gasoline and the potentially different toxic properties of Mn in different forms. PMID- 10385910 TI - Manganese in the context of an integrated risk and decision process. AB - Current approaches to risk assessment regard it as a process that should embody both health and ecological risks, societal values, and cost-benefit analysis, that should seek the views of affected parties, and that should examine available options more holistically than in the past. Even with a single agent, manganese, the process requires a great breadth of information and keen attention to how all of its different components fit together. An evaluation of exposure variables alone needs to consider contributions from multiple media, their physical forms and pathways such as inhaled fumes and particles, and ingestion of water, food, soil, and dust (especially by children). Endpoints need also to be broadened, especially to include susceptibility across the life cycle and the impact of low level neurotoxicity on rate of aging. Finally, the pursuit of risk reduction options for manganese should be embedded in a process that clarifies all), the consequences of a particular option, including the raising or lowering of other risks and the full economic consequences. PMID- 10385911 TI - Genetic and epigenetic control of the proliferation and differentiation of mouse epidermal melanocytes in culture. AB - Serum-free culture of epidermal cell suspensions from neonatal skin of mice of strain C57BL/10JHir (B10) showed that alpha-melanocyte-stimulating hormone (alpha MSH) was involved in regulating the differentiation of melanocytes by inducing tyrosinase activity, melanosome formation, and dendritogenesis. Dibutyryl adenosine 3':5'-cyclic monophosphate (DBcAMP) similarly induced the differentiation of melanocytes. On the other hand, DBcAMP induced the proliferation of epidermal melanocytes in culture in the presence of keratinocytes. Basic fibroblast growth factor (bFGF) was also shown to stimulate the sustained proliferation of undifferentiated melanoblasts in the presence of DBcAMP and keratinocytes. These results suggest that the proliferation and differentiation of mouse epidermal melanoblasts and melanocytes in culture are regulated by the three factors; namely, cAMP, bFGF, and keratinocyte-derived factors. Moreover, serum-free primary culture of mouse epidermal melanocytes derived from B10 congenic mice, which carry various coat color genes, showed that the coat color genes were involved in regulating the proliferation and differentiation of mouse epidermal melanocytes by controlling the proliferative rate, melanosome formation and maturation, and melanosome distribution. PMID- 10385912 TI - An ex vivo study of congenital pigmented nevi in epidermal reconstructs. AB - In order to study morphologic and functional characteristics of pigment cells in congenital pigmented nevi, autologous or heterologous reconstructs have been made using normal keratinocytes and nevus cells from the dermal-epidermal junction or from the dermis. All these cells, keratinocytes and nevus cells, were used as cell suspensions immediately after dissociation from the tissues or after subsequent brief cultivation in a serum-free medium. Reconstructed epidermis were cultured for 15 days at the air-liquid interface with or without ultraviolet (UV) B exposure. The reconstructs were examined macroscopically (formation of hyperpigmented macules), histologically (pigment cell nesting) and ultrastructurally (pigment structure and transfer). Typical nesting of nevus cells was observed in the dermal-epidermal junction or in the superficial dermis associated with macroscopically detectable small pigmented macules. UVB exposure induced an upward migration of nevus cells in the suprabasal layers of the epidermis. This tissue model can be considered as an excellent system for the ex vivo reproduction of pigmented nevi and as an assay of the sensitivity of nevus cells towards UVB irradiation. PMID- 10385913 TI - Horizontal and vertical pigment spread into surrounding piebald epidermis and hair follicles after suction blister epidermal grafting. AB - Following the earlier description of Carnot and Deflandre in 1896, pigment spread phenomenon in mammals was investigated using immunogenetically marked melanocytes (Billingham and Silver, Quart. Rev. Biol. 1960 35: 1-40; Billingham and Silver, Ann. N.Y. Acad. Sci. 1963 100: 348-363). In spite of a number of similar studies on vitiligo lesions, detailed evaluation of pigment spread in piebald lesions has not been reported. To gain further insight into the pigment spread phenomenon in human skin, five piebald patients were studied, on whom suction blister epidermal grafting therapy onto piebald patches was performed. In the present study, pigmentation of all epidermal grafts from normally pigmented areas spread horizontally. It was also found that pre-existing white hairs in recipient sites became pigmented within 1 year after epidermal grafting. Immunofluorescence studies using melanocyte-specific antibody NKI/beteb revealed the newly induced presence of melanocytes in the newly pigmented hair follicles. Further, to study the possible mechanisms inhibiting melanocyte migration from normal skin into piebald lesions, epidermis was grafted from border zones (containing both normal and piebald skin) into the center of hypopigmented lesions. Melanocytes clearly migrated through the border zone of grafted epidermis into surrounding recipient hypopigmented sites. PMID- 10385914 TI - The patchwork mouse phenotype: implication for melanocyte replacement in the hair follicle. AB - Mice homozygous for the recessive patchwork (pwk) mutation are characterized by a variegated pigment pattern with a mixture of unpigmented and normally pigmented hairs. The pigmented hair bulbs contain functional melanocytes. By contrast, the unpigmented hair bulbs contain no melanocytes. This lack results from the death of melanoblasts in the hair follicle at the end of embryogenesis. Here, we report that melanoblasts and melanocytes are found in the epidermis of pwk/pwk mice. Furthermore, these epidermal pigment cells are able to colonize new hair follicles after skin wounding. Despite the presence of epidermal pigment cells with a colonization potential, a follicle that had produced an unpigmented hair produces a new unpigmented hair during the successive hair growth cycles. This hair color continuity is also true for the pigmented hair follicles. Thus, in normal conditions, the hair acts as an independent functional unit as regards its pigment cells population. PMID- 10385915 TI - Microphthalmia-associated transcription factor (MITF) locus lacks linkage to human vitiligo or osteopetrosis: an evaluation. AB - The microphthalmia-associated transcription factor (MITF) locus has been mapped to human chromosome 3p12-p14.1, and encodes a basic helix-loop-helix zipper (bHLH ZIP) protein homologous to a number of transcription factors. Numerous mutations at the mouse microphthalmia (mi) locus have been described, and all have reduced or absent pigmentation of the eyes, ears, and/or pelage, with some genotypes exhibiting small or absent eyes and osteopetrosis. The mivit/vit mutation at the mouse mi locus produces a postnatal depigmentation that resembles human vitiligo. The mice homozygous for this mi allele show a progressive loss of cutaneous, hair and ocular pigmentation with age. Vitiligo, an acquired depigmentary disorder, is characterized by patchy depigmentation of skin that generally begins around puberty and tends to become more progressive over time. There is suggestive evidence that human vitiligo may be inherited; however, the mode of inheritance is still debated and the pathogenesis is not clearly delineated. The human disorder osteopetrosis is characterized by a generalized net accumulation of skeletal mass and results from reduced osteoclast function in the bone. This is an inherited disorder and has been associated with mi in a mutant mouse. Therefore, the possible involvement of the MITF locus in the pathogenesis of either familial vitiligo or osteopetrosis was investigated. Linkage analysis was performed using microsatellite polymorphic markers D3S2465, D3S1261, and D3S1766 on genomic DNA from 26 families with vitiligo/osteopetrosis. D3S1261 is physically located at or near the MITF locus, while D3S2465 and D3S1766 are flanking the locus at about 17.5 cM genetic distance each side. Evidence from LOD score analysis surprisingly indicated that none of the families with vitiligo or osteopetrosis are linked to these short tandem repeat polymorphisms (STRPs). Thus, the human homolog (MITF) of the mouse mi gene, a good candidate gene at the phenotypic level, may not be involved in the pathogenesis of familial human vitiligo or osteopetrosis. PMID- 10385916 TI - The effect of temperature on the light-induced pigment movement in fish corneal chromatophores. AB - Corneal chromatophores of unusual morphology were used for studies on the influence of temperature on the intracellular pigment movement in two species of marine fish from different temperature zones: the tropical puffer, Canthigaster cinctus, and boreal whitespotted greenling, Hexagrammos stelleri. It was shown that both dispersion under bright illumination and aggregation at darkening are slower or decrease at lower temperatures when examined in the range of 12-27 degrees C. The mean speed of the pigment translocations in the individual cell process was 0.38 micron/s at the highest temperature examined, with a range of 0.17-1.0 micron/s. Near the middle of the temperature range, the dynamic characteristics of cell pigment movement in tropical and boreal species were rather close, suggesting that there would be little divergent adaptations with respect to the mechanisms of the pigment transport. Corneal chromatophores are considered as a new promising model for cell motility studies. PMID- 10385917 TI - Effects of fluorescent light on growth of bovine retinal pigment epithelial cells in vitro incubated with linoleic acid or linoleic acid hydroperoxide. AB - Light-induced peroxidation of polyunsaturated fatty acids (PUFA) may generate lipid hydroperoxides, which may have toxic effects on retinal pigment epithelial (RPE) cells in vitro. We investigated the effects of cool-white fluorescent light on the RPE cells incubated with linoleic acids (LA) or linoleic acid hydroperoxides (LHP) and the influence of antioxidative enzymes. We measured the bovine RPE cell number after exposure to fluorescent light (610 and 1,200 lux) in the presence of LA or LHP. Furthermore, the effects of superoxide dismutase (SOD) and catalase on LA- or LHP-treated RPE cells were also examined. Both LA and LHP treatment increased RPE cell number under weak illumination (610 lux), but dose dependently decreased the number of cells exposed to strong illumination (1,200 lux). With exposure to strong illumination, LA caused a greater reduction in RPE cell number than LHP. Multiple linear regression analysis showed that the number of RPE cells was significantly decreased in a manner dependent on the interactions of the illuminance of light and the concentrations of LA or LHP. The antioxidative enzymes significantly ameliorated the damage to RPE cells from LA or LHP and exposure to light. Therefore, the exposure to fluorescent light augmented the cytotoxic effects of LA and LHP on RPE cells, and this effect is likely to be mediated by reactive oxygen species. PMID- 10385918 TI - beta-Adrenergic receptor subtypes in melanophores of the marine gobies Tridentiger trigonocephalus and Chasmichthys gulosus. AB - The subtype of beta-adrenergic receptors in melanophores of the marine gobies Tridentiger trigonocephalus and Chasmichthys gulosus was studied. Pigment of denervated melanophores in isolated, split caudal fins was preliminarily aggregated by incubating the specimens in a physiological saline containing 10 microM phentolamine and 30-100 microM verapamil or 2-10 nM melatonin, and the responses of the melanophores to a beta-adrenergic agonist added to the incubating medium were recorded photoelectrically. The beta-adrenergic agonists noradrenaline, adrenaline, isoproterenol, salbutamol and, dobutamine were all effective in evoking a dispersion of melanophore pigment in the presence of phentolamine and verapamil or melatonin. The pigment-dispersing effect of noradrenaline (beta 1-selective agonist) was inhibited by metoprolol (beta 1 selective antagonist), propranolol,- and butoxamine. Whereas, the effect of salbutamol (beta 2-selective agonist) was hardly inhibited by metoprolol, though it was considerably inhibited by propranolol and ICI-118551. It was estimated that beta 1- and beta 2-adrenergic receptors coexist at ratios of 8.6:91.4, in the melanophore of Tridentiger trigonocephalus, and 25:75, in the melanophore of Chasmichthys gulosus, through the analyses of Hofstee plots of the effects of the beta-adrenergic drugs. It was suggested that the relation between the pigment dispersing effect of a beta-adrenergic agonist on the melanophores and the concentration of the drug follows mass action kinetics, when the effect is mainly caused by the activation of beta 2-adrenergic receptors of the melanophores. However, when it is mainly caused by the activation of beta 1-adrenergic receptors of the melanophores, the relation does not follow mass action kinetics. PMID- 10385919 TI - New vistas in immunopsychopharmacology. AB - The fifties of this century were a time of expansion of psychophysiology, psychopharmacology as well as immunology. But the immune system was considered by several years as completely autonomous, indetifying foreign elements to the host, and clearing them from the body. Selye was the first who put forward the hypothesis that the thymus takes part in stress phenomena and the pituitary adrenal-endocrine system mediates the relations between stress and immune system. Then many reports on the immunological consequences of behavior were published. Herbert Novera Spector coined the term neuroimmunomodulation (NIM), and used it in the paper published in 1983. First International Workshop on NIM was held in 1985. Advances in research of the past decade brought psychoimmunopharmacology out of the folklore into the science. The perspectives of the development of understanding further details of NIM depend upon application of antisense nucleotides in further experiments, estimation of cytokine level in cerebro spinal fluid, observations of central effects in patients treated by cytokines, measurements of several immunological, behavioral biochemical and neuroendocrine variables. PMID- 10385920 TI - Normal and pathological distribution of nitric oxide in the cardiovascular system. AB - Using microsensors, it is possible to quantify the amount and concentration of nitric oxide (NO) release throughout the cardiovascular system in veins, arteries and the heart. Under normal physiological conditions a well defined distribution of NO is maintained. This concentration depends++ on the laminar, turbulent, or pulsatile flow rate of blood. Significantly reduced production of NO is observed in the pathogenesis of cardiovascular disorders like hypertension, atherosclerosis and diabetes. This is due to increased generation of superoxide by a dysfunctional endothelium and the rapid formation of peroxynitrite followed by formation of peroxynitrite followed by the formation of highly reactive OH and NO2 radicals and NO2+. Elevated concentration or improved mass transport of L arginine and (6)-5,6,7,8-tetrahydrobiopterin can be applied to increase/decrease NO/superoxide release by the dysfunctional endothelium. PMID- 10385921 TI - Endogenous drug-like factors. AB - The discovery of opioid receptors and endogenous substances capable of specific binding to these receptors, i.e. endorphin and enkephalin, is one of the most spectacular indications suggesting that the presence of a receptor for a certain drug in the organism authenticates searching for an endogenous substances with high affinity at this receptor. Later, further studies were undertaken to detect other endogenous drug-like factors. Some experiments led to the discovery of digoxin-like factor in blood which could bind to a specific receptor on Na+, K(+) ATPase subunit, showing also the affinity for cardiac glucosides. Digoxin-like factor was detected in blood of healthy people who did not receive any drug treatments. It has been estimated to be present in 15% of the population but in some diseases this value is much higher, e.g. digoxin-like factor was detected in 90% of patients with IDDM, and it can be used as a risk factor of the occurrence of vascular complications. In cases with NIDDM, the digoxin-like factor was detected in patients with insulin-resistance. The presence of digoxin-like factor was ascertained in patients with heart failure who did not take digitalis preparations. Endogenous digoxin-like factor can contribute to the detection of falsely increased digoxin blood concentrations during the monitoring of drug level in the course of the therapy. In our studies we ascertained the presence of the quinidine-, cyclosporin-, theophylline- and phenytoin-like substances in the blood of the healthy people who did not receive any drugs. It seems that these endogenous substances resembling drugs are synthesized in human organism when they are needed for maintaining the physiological equilibrium. We can suggest that stimulation of the production of drug-like factors in the organism can be used in the future in the therapy of some diseases. PMID- 10385922 TI - Effect of local intracerebral administration of EMD 57445, a selective sigma receptor ligand, on the locomotor activity of the rat. AB - EMD 57445 is a new compound which has been characterized by high affinity for sigma receptor sites. It has not been shown to bind to any other receptors, including dopamine ones. However, hitherto existing data (behavioral as well as biochemical) have suggested that this drug exhibits functional antidopaminergic activity. Therefore, in the present study, the local cerebral administration of EMD 57445 has been used in order to better elucidate the actual site of action of this compound in the central nervous system. EMD 57445 given unilaterally into the nucleus accumbens, striatum and lateral ventricle decreases the locomotor activity of the rats, the effect being the most pronounced in the case of administration into the nucleus accumbens. Moreover, unilateral intraaccumbal injection of EMD 57445 significantly diminishes apomorphine (given peripherally) induced hyperactivity. Local administration of EMD 57445 into the prefrontal cortex of the rats also attenuated the locomotor activity but the effect was statistically significant only after bilateral administration of the compound. Additionally, the lack of influence of EMD 57445, administered po, on the dopamine D1 and D2 receptor binding in the striatal membranes was observed in these studies. However, in limbic forebrain the density of D2 receptors decreased after the higher dose of EMD 57445. In conclusion, the results obtained in this paper support the hypothesis that EMD 57445 exerts antidopaminergic activity through indirect inhibition of dopamine system. PMID- 10385923 TI - Differential effects of intrathecally and intracerebroventricularly administered nitric oxide donors on noxious mechanical and thermal stimulation. AB - Involvement of nitric oxide (NO) in nociceptive transmission is well documented. However, there is controversy concerning the exact role of NO in mediation of nociception at different levels of the nervous system. Most studies agree that NO promotes hyperalgesia at the level of the spinal cord. Conversely, at supraspinal sites exogenously applied NO has been found to be both pro- and antinociceptive. In light of this discrepancy, the aim of the present study was to compare the effects of NO donors on nociceptive transmission at spinal and supraspinal sites of the central nervous system using mechanical (paw pressure; PP) and thermal (tail-flick; TF) noxious stimulation. Four NO donors which release NO through different mechanisms were used: S-nitrosoglutathione (SNOG; 3-600 nmol), S nitroso-N-acetylpenicillamine (SNAP; 0.18-4.5 nmol), hydroxylamine (HYD; 60-1200 nmol) and 3-morpholino-sydnonimine (SIN-1; 490-970 nmol). They were injected intrathecally (i.t.) or intracerebroventricularly (i.c.v.) to male Wistar rats and nociceptive thresholds were evaluated in TF and PP tests. It was found that NO donors administered i.t. or i.c.v. produced a dose-dependent hyperalgesia in the PP test. The hyperalgesia induced by mechanical stimuli was stronger after i.t. than after i.c.v. administration of NO donors. The SIN-1-induced hyperalgesia, as evaluated by teh PP test, was reversed by i.t. pretreatment with haemoglobin (1.5-4 nmol) a NO scavenger, and methylene blue (267-1070 nmol) a guanylate cyclase and NO synthase inhibitor, suggesting that NO exerts its action by facilitating cyclic guanosine 3',5'-monophosphate (GMP) formation. Unlike in the PP test, SNAP and SNOG had no effect on the nociceptive threshold in the TF test, and only SIN-1 administered i.t. produced a weak hyperalgesia in that test, while HYD caused a mild but significant prolongation of the TF reflex. The above data show that NO produces hyperalgesia principally in response to noxious mechanical stimuli. This effect seems to be predominantly mediated in the spinal cord, however, it occurs at both levels of the central nervous system. PMID- 10385924 TI - Modulation of neutrophil activity by submandibular gland peptide-T (SGP-T). AB - The heptapeptide submandibular gland peptide-T (SGP-T; sequence = Thr-Asp-Ile-Phe Glu-Gly-Gly) was isolated from rat submandibular glands based on its ability to reduce endotoxic hypotension. Since these glands also modulate neutrophil function, the effects of SGP-T on neutrophil function were investigated. I.v. SGP T (35 and 100 micrograms/kg), when administered prior to and after the s.c. implantation of a carrageenan soaked sponge into rats, significantly reduced the accumulation of neutrophils in the sponge. Neutrophils retrieved from saline soaked sponges generated substantial amounts of superoxide anion in response to both phorbol myristate acetate (PMA) and N-fornyl-methionyl-leucyl-phenylalanine (fMLP), whereas those obtained from carrageenan impregnated sponges were refractory to these oxidative stimuli. Treatment with SGP-T promoted a dose dependent recovery in the ability of neutrophils obtained from carrageenan soaked sponges to generate superoxide anion. This study, by identifying SGP-T as a regulator of neutrophil function, supports the concept that salivary glands are involved in the regulation of inflammatory responses. PMID- 10385925 TI - Postmortem instability of dopamine and its metabolites in rat striatum and limbic forebrain. AB - The level of dopamine (DA) and its metabolites 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA) and 3-methoxytyramine (3-MT) were determined in the brains of rats kept 24 h after death at two different temperatures, 4 degrees C and 22 degrees C. The estimations were carried out in the striatum and limbic forebrain containing: nucleus accumbens, septum, limbic cortex, amygdala, tuberculum olfactorium. Brain tissue of control rats was dissected immediately after decapitation, frozen over solid CO2 and stored at -70 degrees C until assayed. DA and its metabolites were measured, using high-performance liquid chromatography (HPLC) with electrochemical detection. The levels of DA, DOPAC and HVA in the striatum were significantly decreased (from 50% to 80%) when rats were kept 24 h after death. The changes were more pronounced at 22 degrees C than at 4 degrees C. As the decrease in DA concentration was stronger than that of its final metabolite HVA, the ratio of HVA/DA concentration measured as an index of the rate of DA metabolism was even increased (from 8 to 11). Different changes occurred in the limbic region, where the levels of DA and HVA did not change neither at 4 degrees C nor 22 degrees C. The level of intraneuronally formed DA metabolite-DOPAC was elevated (by about 60%). The level of 3-MT, extraneuronally formed DA metabolite, was significantly increased both in the striatum (200%) and limbic DA structures (500%). These data demonstrate regional postmortal differences in stability of DA and its metabolite levels, which are in the striatum temperature-, time-, and storage-dependent. That implicates a careful assessment of postmortem studies when measuring the neurotransmitter dynamics in human necropsy material. PMID- 10385927 TI - Reserpine induces increase in neuropeptide Y immunoreactivity in rat amygdala neurons. AB - The effect of reserpine, a monoamine-depleting agent, on the neuropeptide Y (NPY) immunoreactivity was studied immunohistochemically in the amygdaloid complex (amygdala) of rat brain. It was found that reserpine, at a dose of 10 mg/kg i.p., after 24 h, enhanced the NPY immunoreactivity in amygdala neurons, which was expressed as an increase in the staining intensity and in the number of immunoreactive neurons visible in that structure. The obtained results suggest that monoamines take part in the regulation of NPY expression in amygdala neurons of rat brain, and that elimination of these monoaminergic regulation leads to an increase in the NPY content in that structure. PMID- 10385926 TI - Pharmacokinetics of phenothiazine neuroleptics after chronic coadministration of carbamazepine. AB - The aim of the present study was to assess the influence of carbamazepine on the pharmacokinetics of the two phenothiazine neuroleptics thioridazine and perazine in rats. The obtained results are compared with the results of analogical experiments concerning promazine. Thioridazine or perazine (10 mg/kg i.p.) were administered twice a day for two weeks alone or jointly with carbamazepine (15 mg/kg i.p. during the 1st week, and 20 mg/kg i.p. during the 2nd week of treatment). Concentrations of the neuroleptics and their main metabolites in the plasma and brain were measured at 30 min, 6 and 12 h after the last dose of the drugs. Carbamazepine decreased the concentrations of thioridazine and its metabolites (especially mesoridazine and sulforidazine) in plasma at 30 min and 6 h after the last dose of the drugs. Similar changes in the concentrations of thioridazine and its metabolites were observed at 6 h in the brain. Carbamazepine did not significantly influence the pharmacokinetics of perazine. In vitro studies with liver microsomes of control rats revealed that carbamazepine added to the incubation mixture inhibited N-demethylation of thioridazine via mixed mechanism, but it did not influence significantly 2- or 5-sulfoxidation of the neuroleptic. In the case of perazine, no distinct inhibition of its N demethylation or sulfoxidation by carbamazepine was observed. Neither carbamazepine nor the neuroleptics, administered separately or jointly for two weeks, significantly influenced the concentrations of cytochromes P-450 and b-5 in the liver. Carbamazepine++ given chronically decreased the rate of N demethylation and had a tendency to accelerate 2-sulfoxidation of thioridazine, both when given alone (as compared to the control) and when coadministered with thioridazine (as compared to the thioridazine-treated group). In contrast, chronic treatment with carbamazepine alone, significantly increased the rate of perazine N-demethylation. When carbamazepine was coadministered with perazine, the effect was less pronounced. In conclusion, carbamazepine given jointly with thioridazine or promazine at pharmacological doses to rats accelerates the metabolism of the neuroleptics, which is not the case with perazine. The observed induction proceeds by metabolic pathways other than N-demethylation or sulfoxidation. The different effect of carbamazepine on the N-demethylation of thioridazine and perazine in liver microsomes of control and carbamazepine treated rats implicates that the two reactions are not catalyzed by the same enzyme. Such an induction of neuroleptic metabolism by carbamazepine in patients may worsen psychotic symptoms. PMID- 10385928 TI - Opposite effects of inhibitory and excitatory neurosteroids on [3H]dopamine release from rat nucleus accumbens. AB - Neurosteroids with GABAA receptor antagonistic properties increase K(+)-evoked [3H]dopamine release from rat nucleus accumbens slices, whereas neurosteroid positive modulators of GABAA exert an opposite effect. PMID- 10385929 TI - Mechanical reactivity of isolated human and guinea-pig portal veins to spasmogens. AB - The pattern of mechanical reactivity to prostaglandin F2 alpha and endothelin-1 was different in circular muscle preparations of the extra-hepatic portal vein of humans and guinea-pigs. PMID- 10385930 TI - Nosology and epidemiology of addictive disorders and their comorbidity. AB - Diagnostic classification systems have developed to the point at which the DSM-IV and ICD-10 are nearly identical, so that researchers and clinicians in different parts of the world have a common language for substance-dependence diagnoses. Despite the differences in nosology, the demographic correlates of alcohol and drug dependence are strikingly similar. Lifetime and 12-month prevalences are generally higher in men than in women, whites compared with nonwhites, younger compared with older cohorts, those with lower income levels and lower educational attainment, and those who have not been stably married (including those who have cohabited). NCS data indicated differences in risk factors for stages of drug use, arguing for separation of these in future analyses. Alcoholics are more likely to have another psychiatric disorder compared with their nonalcoholic counterparts, and ASPD, mania, and other drug dependence rank among those disorders most strongly associated with alcohol and drug dependence. Analyses from the NCS examining temporal ordering of diagnoses have focused attention on anxiety disorders in the cause of alcohol dependence. Depression, although not so strongly associated with substance dependence as clinical studies had led researchers to believe, nonetheless seems to be of etiologic interest as well, according to the NCS analyses. The cross-sectional results of the NLAES data on comorbidity between depression and alcohol and drug dependence have uncovered important new associations between gender, age, and depression and may yield further etiologic insights when age-of-onset data are taken into account. PMID- 10385931 TI - Gender differences in substance use disorders. AB - Despite the fact that the rate of substance abuse and dependence is higher among men than it is among women, the prevalence rates, especially the more recent ones, indicate that a diagnosis of substance abuse is not gender specific. From the emerging literature on gender differences over the past 25 years, male and female substance abusers are clearly not the same. Women typically begin using substances later than do men, are strongly influenced by spouses or boyfriends to use, report different reasons for maintaining the use of the substances, and enter treatment earlier in the course of their illnesses than do men. Importantly, women also have a significantly higher prevalence of comorbid psychiatric disorders, such as depression and anxiety, than do men, and these disorders typically predate the onset of substance-abuse problems. For women, substances such as alcohol may be used to self-medicate mood disturbances, whereas for men, this may not be true. Although these comorbid disorders might complicate treatment for women, women are, in fact, responsive to treatment and do as well as men in follow-up. Gender differences and similarities have significant treatment implications. This is especially true for the telescoping phenomenon, in which the window for intervention between progressive landmarks is shorter for women than for men. This is also true for the gender differences in physical and sexual abuse, as well as other psychiatric comorbidity that is evident in female substance abusers seeking treatment. The barriers to treatment for women are being addressed in many treatment settings to encourage more women to enter treatment, and family and couples therapy are standard therapeutic interventions. Negative consequences associated with substance abuse are different for men and women, and gender-sensitive rating instruments must be used to measure not only the severity of the problem but also to evaluate treatment efficacy. To determine whether gender differences observed over the past 25 years become less demarcated in comparisons of younger cohorts of substance abusers in the future will be interesting. Changing societal roles and attitudes toward women, the increase in women entering the workplace, in general, and into previously male-dominated sports and professions, in particular, may influence not only opportunities to drink but also drinking culture. Some gender differences likely will remain, but other gender differences will probably also emerge. The comparison of male and female substance abusers promises to be a fruitful one for researchers. The translation if the research findings to the treatment community to improve treatment outcome for both sexes will be an equally exciting challenge for the field. PMID- 10385932 TI - The role of cultural and social factors in the cause of addictive disorders. AB - For many centuries, generations of young people were protected from the early onset of addictive disorders. Although addiction to drugs and alcohol had been well known for centuries, widespread addiction has occurred only in recent centuries. Because the human gene pool or human biochemistry did not likely change suddenly to produce this result, social and cultural factors likely have produced widespread addiction. From another perspective, the sociocultural factors that once protected our societies against widespread addiction may have become weakened or inoperative. Our social institutions--our families, schools, religions, neighborhoods, and governments--no longer protect us and our young from addiction as they once did. The failure of traditional social institutions to protect us from addiction does not mean that we must seek drug panaceas only in nonsocietal venues, such as medications and psychotherapies. Rather, we should look to those elements of our institutions that have failed us and seek to bolster them. A gradually evolving body of literature on this topic demonstrates that institutional changes can serve to reduce widespread addiction among us. Moreover, these changes can be implemented at many levels: within our families, schools, friendship groups, workplaces, churches, neighborhoods, and legislatures. PMID- 10385933 TI - Addictive disorders in adolescents. AB - Physicians should recognize the importance of individual differences in the etiologic pathway to drug abuse. Drug use in most adolescents subsides or stops by adulthood; however, adolescents with behavioral or affective dysregulation, poor social skills, a limited social network, and substance abuse during late adolescence are at increased risk for substance dependence in adulthood. Research is needed, however, to clarify the developmental emergence and interaction between individual and contextual risk factors. Understanding person-environment processes within a developmental perspective not only yields a better understanding of the causes but also informs about taxonomy, prevention, and readiness to change and compliance in treatment and after-care. Treatment outcome research suggests that (1) relapse is likely to occur within the first 3 months after treatment completion and, to a lesser extent, over the year following treatment completion; (2) relapse is more likely in adolescents who have comorbid psychiatric disorders and other problems, such as high stress, low social skills, lack of involvement in productive activities or active leisure, and no follow-up intervention; (3) continued after-care treatment may maintain treatment gains; (4) the effectiveness of treatment and aftercare is likely to vary by the amount, mode, and the consistency with which it is delivered; (5) gender differences might have an impact on treatment outcome; and (6) adolescents presenting for treatment are likely to respond well to interventions based on family therapy and CBT approaches. PMID- 10385934 TI - Genetics of alcoholism and substance abuse. AB - Twin studies have demonstrated that addictive disorders are genetically and environmentally influenced. Our knowledge of behavioral differences predisposing to addiction is advancing rapidly, particularly in alcoholism but also in the other addictions, through studies on animals and humans. Recently, linkage analyses in humans and rodents have pointed to genomic regions harboring genes which influence addiction or drug-associated behaviors. There is increasing evidence that the addictions have common as well as distinct neurobiological pathways. These advances in the understanding of the genetics of addictive disorders should facilitate the development of specific pharmacotherapies and the more accurate targeting of therapies using molecular diagnostic approaches. PMID- 10385935 TI - Neurobiology of tobacco smoking and other addictive disorders. AB - Advances in research of the neurobiology of addictive disorders have provided clinicians with an evolving perspective on addiction. All drugs of abuse seem to share a common neurobiologic substrate involving the mesocorticolimbic system. Considerable evidence shows that these dopaminergic projections are involved in the positive brain reward, which drives addictive disorders; however, recent studies also implicate the neurotransmitters glutamate and serotonin in learning and sensitization to drug use. A review of the neurobiology of tobacco smoking provides further examples of the mechanisms for reinforcing tobacco use, including the enhancement of memory and treatment of depression with nicotine and MAO-inhibiting chemicals in tobacco smoke respectively. The relevance of these advances may be realized through the destigmatization of addictive disorders and the development of new and improved treatment strategies. PMID- 10385936 TI - Structural and functional neuroimaging findings in substance-related disorders. AB - Intoxication with alcohol results in depressed global glucose metabolism that continues into the stages of withdrawal and abstinence. The decrease in metabolism, however, is not equal across the brain, with certain regions more affected than others. Such a pattern of disturbance suggests that the effect of alcohol on the brain cannot simply be a nonspecific depressant effect secondary to decreased blood flow or glucose transport into the cells but may be related to the dysfunction of the various neurotransmitter systems. Different authors have suggested the dysfunction to be related to the GABAergic, cholinergic, and dopaminergic systems. Long-term alcoholism is associated with atrophy of several brain regions. The frontal lobes and limbic structures seem to be most vulnerable. The data are encouraging with regard to the normalization in brain metabolism and in size of vulnerable brain regions with continued abstinence. In addition to findings of improvement in cognitive functioning and many health parameters, these findings arm clinicians with further data on the benefits of abstinence in the struggle to aid patients in maintaining their sobriety. Several areas remain to be addressed. In particular, clinicians are in need of data, neuroimaging and otherwise, that serve as prognostic indicators, thus allowing patients at higher risk for relapse to be identified and provided with more intensive treatment. A similar need exists for indicators of diagnostic heterogeneity that would guide the development of more highly tailored treatment regimens for identified subgroups of patients. Currently, we have rudimentary knowledge of the gender differences of the effects of alcohol and cocaine on the brain. PMID- 10385937 TI - Overview of psychiatric comorbidity. Practical and theoretic considerations. AB - Psychiatric comorbidity is a common clinical challenge for addiction therapists. The clinician is challenged by the assessment as well as the ongoing management of such cases. Stabilization is complicated by the comorbid problems. Careful use of psychiatric medications can be helpful and collaboration with other health care professionals is usually an important aspect of treatment with this population. There is much evidence that treatment of addiction is cost-effective and some evidence that treatment of dual conditions is also cost-effective. PMID- 10385938 TI - Medical consequences of substance abuse. AB - Given the preceding review, differentiating the complications of parenteral drug use, HIV disease, and the toxicity of the drugs such as alcohol or cocaine may be a difficult matter for clinicians. The risk for coexisting morbidities is high. Thus, obtaining accurate and complete medical histories is of paramount importance. Drug-abuse treatment and follow-up medical care after an acute complication often involves multiple health care providers. The integration of primary prevention plans with the reinforcement of drug abstinence requires time, commitment, and the coordination of services. This integration should be a priority for individual patients as well as for public health planning. PMID- 10385939 TI - Mortality risks in alcoholism and effects of abstinence and addiction treatment. AB - The mortality rate from alcoholism and related comorbidities is high. Studies show multiple causes of premature death from alcoholism. Several studies showed that abstinence had a positive effective on the overall survival of alcoholics. Alcoholics who abstained from alcohol, particularly continuously, showed reduced mortality rates and increased years of longevity than alcoholics who relapsed to alcohol consumption. The sources of the findings tend to be derived from treatment populations, in which abstinence is expected to occur in higher rates than in the general population. PMID- 10385940 TI - Cost-effectiveness of substance abuse services. Implications for public policy. AB - Cost-effectiveness analysis, a technique for allocating resources, examines the relationship between the cost of providing treatment and resulting improvement in health measured in a single, numerical scale. In applying this concept to substance abuse services, the authors expressed effectiveness in terms of additional "abstinent years." To control for differences in clients across modalities, the authors used multivariate cost-effectiveness analysis, estimating results for a typical client at each of three alternative severity levels. PMID- 10385941 TI - Pharmacologic treatments for drug and alcohol dependence. AB - Pharmacotherapy remains a relatively underused strategy for the treatment of patients with psychoactive substance-use disorders. This is partly because of a widely held view among the public and the substance-abuse treatment community that substance-use disorders are non-medical and should be treated through nonpharmacologic means. Furthermore, the pharmaceutical industry has been slow in developing medications for the treatment of patients with these disorders because of skepticism over the potential profitability of such medications. These factors have limited research activity in this area, with much of the impetus for study of these medications coming from the National Institute on Drug Abuse and the National Institute on Alcohol Abuse and Alcoholism, where the substantial public health implications of medications development for substance-use disorders have been recognized. Despite limitations, considerable advances in medication development for patients with substance-use disorders have occurred in recent years, and more can be expected in the near future. This is most evident in the treatment of patients with nicotine and opioid dependence, for whom several pharmacologic options exist. Recently renewed interest in the pharmacologic treatment of patients with alcohol dependence is likely to advance that therapeutic area substantially with time. Ongoing efforts should focus on identifying new compounds, systematically assessing them for activity in specific substance-use disorders, and educating the public and the treatment community about the substantial benefits that can accrue from medication development for patients with substance-use disorders. PMID- 10385942 TI - Twelve-step and mutual-help programs for addictive disorders. AB - Psychiatrists may wonder why both addiction treatment and the 12-step programs recommend abstinence. In his 50-year follow-up of two groups of alcoholics, Vaillant compared those who established secure abstinence with those who continued to drink. Secure abstinence was associated with: Living longer Better mental health Better marriages Being more responsible parents Being successful employees In considering the various routes to secure recovery, Vaillant recommended that clinicians: Offer the patient a nonchemical substitute for alcohol Remind the patient ritually that even one drink can lead to pain and relapse Repair the social and medical damage that the patient has experienced Restore the patient's self-esteem The preponderance of the research data now available indicates that the 12-step programs of AA, NA, Cocaine Anonymous, and Al-Anon are most helpful for alcohol-dependent and other drug-addicted patients as they seek to achieve secure, long-term abstinence. A growing number of clinicians is recommending that physicians become more knowledgeable and skilled in referring and supporting patients in working 12-step programs of recovery. Specific recommendations include: 1. Be familiar with 12-step activities and tools. These include meetings, home groups, sponsors, the Twelve Steps and Twelve Traditions, books, pamphlets, and slogans. To be able to discuss the meanings and applications of these tools for recovery is useful. Physicians can select those that are most suitable for the individual, recognizing that meeting attendance might not be the most important activity. 2. Support referral by facilitating a meeting between the patient and a temporary contact from the 12-step program. This means becoming familiar with local 12-step programs. Phoning the local AA or NA central office or hot line makes connecting patients to someone who will take them to a meeting that same day possible. AA and NA have committees whose members are interested in working with physicians to help get patients to meetings and to get information to physicians. These are the Cooperation with the Professional Community, Treatment Facilities, and Hospitals and Institutions committees. 3. Work with the resistance of patients. Many addicted patients are resistant to the idea of attending 12-step or mutual-help programs. Reminders of their painful personal database associated with the use of alcohol or other drugs can help break through denial. Involvement of family members and friends in the network therapy developed by Galanter can be effective in reducing resistance. Being patient and persistent in developing the therapeutic alliance helps to maintain contact during the first difficult year of recovery. Physicians should be prepared to work with patients as long as necessary to stabilize their sobriety. Zweben has suggested ways psychotherapy can help deepen work with the steps. 4. Help dual diagnosis patients understand AA's and NA's singleness of purpose. These programs work only with addiction; they do not try in any way to deal with other mental disorders. All patients have to say is, "I want to stop drinking or using drugs," and they will be welcomed and accepted at meetings (see Tradition 3). If they talk only about their psychiatric symptoms or medications, someone may suggest that they go elsewhere for help. Occasionally, well-intentioned AA or NA members tell patients to stop taking their medications. The authors always direct patients to the pamphlet The AA Member: Medications and Other Drugs. This pamphlet tells AA members not to play doctor and to take the medications their doctors prescribe. Copies of the pamphlet are widely available at many AA meetings, or they can be ordered by physicians from Alcoholics Anonymous World Services, General Service Office, Box 459, Grand Central Station, New York, NY 10163 (212-870-3400). 5. Get comfortable with the spiritual dimensions of healing. Zweber and Brown offer good suggestions for getting com PMID- 10385943 TI - Coping and social skills training for alcohol and cocaine dependence. AB - Coping and Social Skills Training (CSST) has been in the forefront of the addictions literature for over two decades. In this review, we provide an outline of empirically validated CSST assessment measures and treatment protocols for alcohol dependence and cocaine dependence. Key elements covered in CSST include communication skills training, cue exposure treatment with urge coping skills, and cocaine-specific coping skills training. We conclude with a summary of the research to date in support of CSSt and future directions for the treatment of addictive behaviors. PMID- 10385944 TI - Education and training in addictive diseases. AB - Addiction to alcohol and other drugs is a serious public health problem that is one of the most common disorders seen in medical practice. Although it is an extremely common disorder, it is poorly diagnosed and treated by physicians. Training about addictions must begin early in the medical student's career and continue in a vertically integrated way throughout medical school and residency. The notion of addiction as a disease process must be introduced and integrated into course materials in the preclinical years. Careful attention to the development of positive views toward working with addicted patients must be paid, and students must be indoctrinated early with the idea that physicians have a responsibility to diagnose and manage addicted patients. PMID- 10385945 TI - Postgraduate education and training in addiction disorders. Defining core competencies. AB - The rising prevalence of substance abuse disorders in the general medical population makes it essential that more active training be initiated during general residency training across all specialties. Core competencies for general residencies are outlined and the need for advocacy for continuation of postgraduate fellowship in Addiction Psychiatry is recommended as a means of increasing the numbers of faculty to provide leadership teaching and research in the area of Addiction Psychiatry. PMID- 10385946 TI - The role of the physician in addiction prevention and treatment. AB - With increasing pressure on general physicians by managed care organizations and the public to treat and advocate for drug and alcohol addicted patients, it is more necessary than ever that physicians have the knowledge and skills to appropriately address this segment of the population. Specifically, physicians need a better understanding of the prevalence of alcohol and drug dependence in a variety of populations, along with increased awareness of the economic impact of addictive illnesses on our society. Routine screening questions should be incorporated into patient encounters, and physicians should be able to identify environments that may pose a risk for the development of addiction. Physicians need training and practice in referring patients to treatment teams, monitoring patients in recovery, and providing interventions that will eliminate or reduce substance abuse before it becomes addiction. The treatment outcomes in abstinence based programs, particularly those combined with referral to AA, have been encouraging, demonstrating that addiction is a treatable illness and not a character defect. In addition, several studies provide evidence that addiction treatment is cost-beneficial, resulting in reduced medical costs, lowered absenteeism, and increased productivity. Despite these encouraging results, there is still room for improvement. Treatment is not always effective, and it is not sufficiently available to everyone who needs it. Addicted individuals are both stigmatized and marginalized, and many are too ill to advocate for themselves. Widespread recognition in the medical community of addiction as a treatable illness will contribute to a greater understanding of addictive disorders and reduce the stigma attached to the diagnosis and treatment of addiction. For this to occur, better training for physicians in the recognition and management of addictive disorders, starting at the medical school level, is necessary. The approval of addiction medicine as a clinical specialty by the American Medical Association also has helped to advance the legitimacy of addiction as a treatable illness, and provides a focal point for the synthesis and integration of clinical, teaching, and research activities central to addiction medicine. The combination of knowledge, skills, and attitudes outlined in the article will go a long way toward increasing physicians' abilities to assist their patients with recovery from addiction. PMID- 10385947 TI - History of intraoperative ultrasound. AB - Intraoperative ultrasound (IOUS) using A-mode or non-real-time B-mode imaging started in the 1960s; however, it was not widely accepted mainly because of difficulty in image interpretation. In the late 1970s, IOUS became one of the topics in the surgical communities upon the introduction of high-frequency real time B-mode ultrasound. Special probes for operative use were developed. In the 1980s, all over the world the use of IOUS spread to a variety of surgical fields, such as hepatobiliary pancreatic surgery, neurosurgery, and cardiovascular surgery. IOUS changed hepatic surgery dramatically because IOUS was the only modality that was capable of delineating and examining the interior of the liver during surgery. After 1990, color Doppler imaging and laparoscopic ultrasound were incorporated into IOUS. Currently, IOUS is considered an indispensable operative procedure for intraoperative decision-making and guidance of surgical procedures. For better surgical practice, education of surgeons in the use of ultrasound is the most important issue. PMID- 10385948 TI - Three-dimensional ultrasound imaging. AB - The objective of this article is to provide scientists, engineers and clinicians with an up-to-date overview on the current state of development in the area of three-dimensional ultrasound (3-DUS) and to serve as a reference for individuals who wish to learn more about 3-DUS imaging. The sections will review the state of the art with respect to 3-DUS imaging, methods of data acquisition, analysis and display approaches. Clinical sections summarize patient research study results to date with discussion of applications by organ system. The basic algorithms and approaches to visualization of 3-D and 4-D ultrasound data are reviewed, including issues related to interactivity and user interfaces. The implications of recent developments for future ultrasound imaging/visualization systems are considered. Ultimately, an improved understanding of ultrasound data offered by 3 DUS may make it easier for primary care physicians to understand complex patient anatomy. Tertiary care physicians specializing in ultrasound can further enhance the quality of patient care by using high-speed networks to review volume ultrasound data at specialization centers. Access to volume data and expertise at specialization centers affords more sophisticated analysis and review, further augmenting patient diagnosis and treatment. PMID- 10385949 TI - Pre-ejectional left ventricular wall motions studied on conscious dogs using Doppler myocardial imaging: relationships with indices of left ventricular function. AB - Duration of the pre-ejection period is a sensitive index of myocardial function. Our purpose was to document normal pre-ejectional left ventricular (LV) wall motions at rest and under dobutamine using Doppler myocardial imaging (DMI), and to correlate posterior wall velocities with indices of LV systolic function. M mode recordings of both walls were imaged on eight conscious dogs chronically instrumented. Subendocardial pre-ejectional velocities were digitized and measured every 3.8 ms. DMI analysis consisted of sign recognition, velocity measurement, duration and timing from the Q wave of the electrocardiogram. Isovolumic contraction time (Ict) was represented by the time interval from onset to peak of the first derivative of LV pressure. Conventional Doppler labelling of velocity signs, positive toward and negative away from the transducer, was applied to the direction of encoded wall motions. For physiological understanding, wall motions of both walls were also labelled inward and outward with respect to the left ventricular cavity center. In each wall, PEP was shown as several colored strips, each strip representing the period of time that the wall was moving in one direction. Changes in velocity sign corresponding to changes in direction of motion were opposed in each wall (p < 0.001), featuring successive inward and outward wall motions. There was a markedly sustained inward motion during Ict. Its velocity amplitude increased with dobutamine. There was a positive correlation between velocities of the inward motion contemporaneous of Ict and ejection fraction (r = 0.72, p < 0.003). Values of Ict respectively drawn from DMI and from hemodynamics were also significantly correlated (r = 0.85, p < 0.007). Thus, the inward motion evidenced by DMI during Ict appears promising to assess myocardial function and effect of drugs. PMID- 10385950 TI - Effect of timing of blood pressure measurement in the assessment of arterial stiffness: the SMART Study. AB - In the assessment of arterial stiffness, pulse pressure is measured. Presently, there is no consensus on how pulse pressure should be measured. Distensibility of the left and right common carotid arteries was measured noninvasively in 224 patients participating in the Second Manifestations of ARTerial disease (SMART) study. Blood pressure was recorded every 4 min, using a semiautomatic oscillometric device. Distensibility coefficients (DC) were calculated with pulse pressure obtained as an average of (A) all measurements during the session; (B) the second, third, and fourth measurement; (C) measurements before and after distensibility assessment; and (D) three measurements nearest to distensibility assessment. Associations of cardiovascular risk factors with the four calculated DCs were evaluated with linear regression analysis. DC estimates were slightly more precise with methods A and B than with C or D. The magnitude of the associations showed a slight trend to higher precision for methods A and B. Pulse pressures obtained as an average of all or the second, third, and fourth blood pressure measurements during an arterial stiffness measurement session yield slightly more precise estimates of DC. However, the differences between the methods are small; therefore, we suggest that pragmatic arguments dominate the choice between the methods. PMID- 10385951 TI - Determination of carotid plaque risk by ultrasonic tissue characterization. AB - This in vitro study investigated the ability of ultrasonic tissue characterization (UTC) to discriminate between plaques from asymptomatic and symptomatic patients and to compare UTC findings with quantitative measurements of plaque morphology. A total of 34 plaque specimens removed at carotid endarterectomy were scanned transversely at intervals of 1 mm, and compared to tissue cross-sections examined by optical microscopy employing computer-assisted planimetry. UTC was performed by spectral analysis of backscattered radiofrequency signals. The slope, intercept and total power parameters of the spectrum were evaluated. Discriminant analysis was used to compare the ability of the UTC spectral parameters and morphological constituents to correctly classify plaques according to their symptom group membership. UTC correctly classified 88.2% of the plaques. Thrombus was present in 93.9% of the plaques, and there was little difference in the morphological constituents of plaques from asymptomatic and symptomatic patients. Morphological constituents correctly classified 60.7% of the plaques. We conclude, in this preliminary study, that UTC can discriminate between carotid plaques from asymptomatic and symptomatic patients with moderate accuracy, despite a similarity in their morphological composition. UTC discrimination is not related to differences in the type or amount of morphological constituents in the plaques. PMID- 10385952 TI - Blood velocity profile in the ductus venosus inlet expressed by the mean/maximum velocity ratio. AB - Mean blood velocity (Vmean) is needed for calculating blood flow and possibly the pressure gradient across the ductus venosus. Interference from low velocities from neighbouring vessels makes the direct Doppler measurement of Vmean unreliable. Therefore, it is suggested that Vmean can be derived more reliably from the maximum velocity (Vmax) once the velocity profile, expressed as the ratio Vmean/Vmax, is known. To determine this ratio, ultrasound was performed in 10 fetal sheep during acute experiments under general anaesthesia to ensure good recording control and optimal insonation. Based on 33 Doppler measurements at the ductus venosus inlet, the ratio Vmean/Vmax was determined to be 0.69 (SD +/- 0.07) regardless of Vmax, pulsatility index, vessel diameter, or angle of insonation. These results confirm the previous prediction based on a computational model that the velocity profile is partially blunted. The equation Vmean = 0.7Vmax is recommended for determining Vmean in the ductus venosus. PMID- 10385953 TI - Color Doppler sonography of normal breasts: detectability of arterial blood vessels and typical flow patterns. AB - In a prospective study, 200 healthy female breasts were examined using color Doppler sonography to study the detectability and the resistive indices (RIs) of arterial vessels. In each breast, we attempted to detect two to three vessels and recorded the frequency spectrum with RI of each vessel. Blood vessels (n = 522) could be demonstrated in 196 (98%) breasts. Continuous diastolic flow (RI < 1) was found in 520 (99.6%) vessels. The mean RI of premenopausal women was 0.64; that of postmenopausal women was 0.70. This difference is highly statistically significant (p < 0.0001), but there is a marked overlap between the RIs of both groups. The variation in RI values of all women (up to 0.45), as well as in the breasts of the same woman (up to 0.31), was considerable. We conclude that modern color Doppler devices permit the detection of blood flow in the breast with regularity. Continuous diastolic flow (RI < 1) is a typical flow pattern. The variations of RI between women, and even for the same woman, are remarkable. The mean RI of premenopausal women is lower than the value for postmenopausal women. PMID- 10385954 TI - Vein graft surveillance with scanhead tracking duplex ultrasound imaging: a preliminary report. AB - A severe arterial occlusion in the leg usually is bypassed by implanting a saphenous vein harvested from the limb. Once implanted, the vein functions well but over time may develop stenoses that may lead to occlusion. In order to detect and correct the stenoses that may lead to graft failure, frequent surveillance of the vein graft is required. A new ultrasound imaging method was developed to display the panoramic view of the vein graft in combination with its blood flow velocity waveform for surveillance. The panoramic view is the projection (ray casting) image of multiple B-mode images with sequential longitudinal view of the vein graft. The velocity waveform also is recorded along the vessel with pulsed Doppler ultrasound. The acquired images and waveforms from the ultrasound scanner are registered individually in three-dimensional space with an electromagnet based position and orientation sensor located on the scanhead. A prominent point on the scar from the surgery is used as the fiducial mark for spatial registration, so that the same lesion in the vein graft can be tracked automatically at each visit for retrospective study. PMID- 10385955 TI - A noninvasive method to estimate pulse wave velocity in arteries locally by means of ultrasound. AB - Noninvasive evaluation of vessel wall properties in humans is hampered by the absence of methods to assess directly local distensibility, compliance, and Young's modulus. Contemporary ultrasound methods are capable of assessing end diastolic artery diameter, the local change in artery diameter as a function of time, and local wall thickness. However, to assess vessel wall properties of the carotid artery, for example, the pulse pressure in the brachial artery still must be used as a substitute for local pulse pressure. The assessment of local pulse wave velocity as described in the present article provides a direct estimate of local vessel wall properties (distensibility, compliance, and Young's modulus) and, in combination with the relative change in artery cross-sectional area, an estimate of the local pulse pressure. The local pulse wave velocity is obtained by processing radio frequency ultrasound signals acquired simultaneously along two M-lines spaced at a known distance along the artery. A full derivation and mathematical description of the method to assess local pulse wave velocity, using the temporal and longitudinal gradients of the change in diameter, are presented. A performance evaluation of the method was carried out by means of experiments in an elastic tube under pulsatile pressure conditions. It is concluded that, in a phantom set-up, the assessed local pulse wave velocity provides reliable estimates for local distensibility. PMID- 10385956 TI - Sonographic observations of the gastroduodenal junction in neonatal piglets. AB - Knowledge of the function of the gastroduodenal junction is important, as changes in its motility are associated with gastrointestinal disorders. Sonographic observations were made of the stomach and duodenum of 19 neonatal piglets, 2-6 d of age. Contractions of the stomach and duodenum were identified clearly; the overall rate of gastric contractions was about 4 min-1. The percentage of contractions in which there was a closure of the terminal pyloric antrum and pyloric canal varied, being 57.2% +/- 4.6% in the first postprandial hour and 43.1% +/- 3.0% in the third. Antegrade flow of digesta principally occurred preceding a closure of the pyloric antrum and canal. During contractions of the pyloric antrum, the torus pyloricus moved caudally to fill the lumen of the pyloric canal. Our sonographic method provided a noninvasive technique for studying the form and function of gastroduodenal motility in the neonate, suitable for investigating factors that alter gastric emptying. PMID- 10385957 TI - Improved investigation of portal-hepatic veins by echo-enhanced Doppler sonography. AB - Microbubbles of air released from a galactose vehicle (Levovist) amplify the intensity of Doppler signals. They survive both pulmonary and systemic capillary passage, leading to echo enhancement in the entire vascular system. The aim of this study was to investigate this agent in patients with liver disease and insufficient Doppler signals. A total of 275 Doppler examinations were performed in 176 patients; 20 of these patients could not be studied conventionally due to bowel gas, obesity or noncompliance. They received Levovist to examine portal or hepatic veins or TIPS patency. Angiography, computed tomography (CT) scan or magnetic resonance tomographic angiography (MRTA) was performed subsequently as a control. After administration of Levovist, portal or hepatic veins and TIPS patency could be unequivocally assessed in 18 of the 20 patients. In two patients, suspected occlusion of the portal vein was disproved because the diagnosis was not confirmed later. Only minor adverse effects were encountered. Echo-enhanced Doppler sonography with Levovist is well tolerated. Further study of the value of Levovist for the assessment of portal-hepatic vessels not amenable to conventional Doppler sonography is justified. PMID- 10385958 TI - Duodenal meal stimulation leads to coeliac artery vasoconstriction and superior mesenteric artery vasodilatation: an intra-abdominal ultrasound study. AB - To evaluate the influence of duodenal feeding on splanchnic blood flow, 14 patients with normal coeliac and superior mesenteric arteries underwent intra abdominal duplex scanning of the systemic and splanchnic circulation under standardised basal and meal-stimulated conditions. Doppler samples and diameter measurements were taken of the aorta, coeliac artery, common hepatic artery, splenic artery, superior mesenteric artery, and inferior mesenteric artery. Duodenal meal stimulation has no systemic effects (p > 0.4). However, duodenal meal stimulation results in coeliac artery vasoconstriction (p < 0.06) and superior mesenteric artery vasodilatation (p < 0.05). This study supports other reported results that gastrointestinal blood flow is dependent on the site of food stimulation. PMID- 10385959 TI - Estimation of the human liver volume and configuration using three-dimensional ultrasonography: effect of a high-caloric liquid meal. AB - The aim of this study was to investigate whether or not a magnetic position sensing system for free-hand acquisition of 3-D ultrasound images could be used to estimate liver volumes, and to study the effect of a high-caloric meal on these volumes in healthy subjects. In vitro accuracy was evaluated by scanning porcine and rabbit livers. Ten healthy subjects were examined fasting and 30 min after ingesting a high-caloric liquid meal. Portal and hepatic vein blood flow were measured by 2-D duplex sonography. The 3-D system yielded a strong correlation (r = 0.99) between true and estimated volumes in vitro. No significant increase in liver volume in response to the meal was seen. However, portal and hepatic vein flow volume increased significantly. Experience in human subjects suggests that a complete 3-D study of liver volumes can be obtained from multiple acoustic windows. In healthy subjects, no significant increase in liver volume was seen in response to ingestion of a high-caloric liquid meal. PMID- 10385960 TI - Automatic three-dimensional reconstruction and characterization of articular cartilage from high-resolution ultrasound acquisitions. AB - Three-dimensional (3D) high-resolution ultrasonography has proved to be useful for in vitro assessment of cartilage remodeling due to osteoarthritis. The diagnosis is performed by computation of the mean thickness of the cartilage, which reveals hypertrophy or thinning, and by 3D reconstruction of the data, which provides essential information about the size, extent, and localization of the lesion. In both cases, preliminary segmention of the cartilage is necessary. This article proposes an algorithm for automatic segmentation of the cartilage from 3D ultrasonic acquisitions of the rat patella, which includes the detection of the cartilage surface and the cartilage/bone interface. The method was designed on the assumption of regularity and smoothness of the interfaces. The use of a global threshold was sufficient to separate the patella area from the background. The cartilage/bone interface was detected by selection of regions of interest (ROIs) encompassing the interface, followed by the detection of the interface within these ROIs using the graph theory. The method was applied to 162 samples. The detection accuracy was judged to be very good or good in 99% of the cases for the cartilage surface and in 86% of the cases for the cartilage/bone interface. The mean cartilage thickness value in the central part of the patella obtained from the automatic detection method was compared to that obtained manually. The coefficient of correlation between the two measurements was 0.92. These results show that our method is reliable. Thus, fast processing of a large number of acquisitions and a more complete analysis of the cartilage surface become possible. PMID- 10385961 TI - Experimental verification of the correlation behavior of analytic ultrasound radiofrequency signals received from moving structures. AB - Conventional pulsed ultrasound systems are only able to detect motion along the ultrasound beam (i.e., axial motion). If the angle between the actual motion direction and the ultrasound beam is known, then the magnitude of the actual motion can be derived. This technique can be applied for laminar blood-flow measurements in straight vessels, but for tissue motion it is inadequate because the local tissue motion direction is unknown and may be position-dependent. Assessment of both the axial motion and the lateral motion (i.e., in the direction perpendicular to the ultrasound beam) makes angle-independent assessment of the magnitude of the actual motion feasible. Information about the axial and lateral motion is available in a set of radiofrequency (RF) signals obtained along the same line of observation (M-mode). The experiments described in the present paper show that axial and lateral motion can be estimated from the shape of the envelope of the 2-D (spatial and temporal) correlation function of analytic M-mode RF signals. Furthermore, it is demonstrated that the shape is also affected by the Band width of the received RF signals, signal-to-noise ratio, and local amplitude and phase characteristics of the ultrasound beam. PMID- 10385962 TI - Evaluation of acoustic properties of the live human smooth-muscle cell using scanning acoustic microscopy. AB - This study was performed to measure the acoustic propagation speed in live human aortic smooth-muscle cells (HASMC), using scanning acoustic microscopy (SAM) and a novel measurement theory that permits the measurement of the acoustic propagation speed in biological samples of unknown thickness. C-mode and X-Z-mode images of HASMC under three different conditions: growing (G); differential (D); and on hypotonic loading (H), were acquired using 100-MHz, 450-MHz and 600-MHz ultrasound. The images exhibit features related to the cell surface curvature and intracellular structure. The theory supporting the methodology is derived in this article and makes use of the interference fringes within the focusing lens of the high-frequency transducer. The propagation speed in the cells was calculated from the location of the interference fringe on the C-mode images and the fringe shift on the X-Y-mode images with 450-MHz ultrasound. The propagation speed in D (1624 +/- 16 m/s) was significantly higher than those in G (1571 +/- 14 m/s, p < 0.05) and H (1585 +/- 8 m/s, p < 0.05). Scanning acoustic microscope measurements, along with the described theory, are useful for studying the acoustic properties of live cells ex vivo and have applications in both pathophysiology and biomechanics. PMID- 10385963 TI - 40-MHZ echocardiography scanner for cardiovascular assessment of mouse embryos. AB - Congenital heart disease results from genetic defects that are manifested at early stages of embryogenesis. The mouse is the preferred animal model for studies of mammalian embryonic development and for an increasing number of human disease models. A number of genes identified in the mouse are critical for normal cardiovascular development, but an understanding of the underlying mechanisms regulating heart development is still incomplete, in part because of the lack of methods to measure hemodynamics in live mouse embryos. We describe the development of a 40-MHz ultrasound scanner, which allows image-guided continuous wave and pulsed Doppler blood flow measurements in mouse embryos, in utero, at the critical early developmental stages. Doppler waveforms acquired from mouse embryonic umbilical vessels, descending aorta, and cardiac ventricles are presented to demonstrate the utility of the method. By combining image-guided ultrasound Doppler with the many available mouse mutants, this approach should lead to new insights into embryonic cardiovascular structure-function relationships. PMID- 10385964 TI - Shear wave elasticity imaging: a new ultrasonic technology of medical diagnostics. AB - Shear wave elasticity imaging (SWEI) is a new approach to imaging and characterizing tissue structures based on the use of shear acoustic waves remotely induced by the radiation force of a focused ultrasonic beam. SWEI provides the physician with a virtual "finger" to probe the elasticity of the internal regions of the body. In SWEI, compared to other approaches in elasticity imaging, the induced strain in the tissue can be highly localized, because the remotely induced shear waves are attenuated fully within a very limited area of tissue in the vicinity of the focal point of a focused ultrasound beam. SWEI may add a new quality to conventional ultrasonic imaging or magnetic resonance imaging. Adding shear elasticity data ("palpation information") by superimposing color-coded elasticity data over ultrasonic or magnetic resonance images may enable better differentiation of tissues and further enhance diagnosis. This article presents a physical and mathematical basis of SWEI with some experimental results of pilot studies proving feasibility of this new ultrasonic technology. A theoretical model of shear oscillations in soft biological tissue remotely induced by the radiation force of focused ultrasound is described. Experimental studies based on optical and magnetic resonance imaging detection of these shear waves are presented. Recorded spatial and temporal profiles of propagating shear waves fully confirm the results of mathematical modeling. Finally, the safety of the SWEI method is discussed, and it is shown that typical ultrasonic exposure of SWEI is significantly below the threshold of damaging effects of focused ultrasound. PMID- 10385965 TI - Vibration sonoelastography and the detectability of lesions. AB - Vibration sonoelastography has been developed for the detection of hard lesions in relatively soft tissue. The basic concept is to propagate low-amplitude and low-frequency shear waves (with displacements below 0.1 mm and frequencies typically below 1000 Hz) through deep organs, and displaying the vibration response in real-time using advanced color Doppler imaging techniques. A hard inhomogeneity, such as a tumor, will produce a localized disturbance in the vibration pattern, forming the basis for detection even when the tumor is isoechoic on B-scan images. This paper focuses on the important quantitative issues concerning the detectability or inherent contrast of lesions. The specific factors of lesion size, relative stiffness and vibration frequency are studied using theoretical models, finite element methods and experimental measurements on tissue-mimicking materials. The results indicate that detectability increases with vibration (shear wave) frequency; however, loss mechanisms ultimately limit the penetration of higher vibration frequencies (in the kHz range). PMID- 10385966 TI - Elastographic imaging of thermal lesions in soft tissue: a preliminary study in vitro. AB - The use of elastography for the visualization of thermal lesions in biological soft tissue in vitro was investigated. Thermal lesions were created in samples of postmortem ovine kidney using a surgical neodymium: YAG (Nd:YAG) laser. The kidney samples were cast in gel, and elastographic images of the lesions were constructed using sonographic information and external markers to locate the region of interest. Gross pathology of the kidney samples confirmed the dimensions of the lesions. Good correlation between the lesion length along the laser fiber axis and maximum diameter measured off of the fiber axis determined from elastographic images and gross pathology photographs was found. PMID- 10385967 TI - Innovative technology for tissue disruption by explosive-induced shock waves. AB - We have developed a novel, less invasive, shock wave source that can be introduced into an arbitrary position in a human body percutaneously. Using this technique we can disrupt cells locally. The shock wave source consists of an explosive, an optical fiber, a balloon catheter, and a Nd:YAG laser, which generates a spherical explosive shock wave. The destructive potential of the present source for injuring tissue was confirmed and the subsequent cell elongation and split in the direction of the shock wave has been observed. PMID- 10385968 TI - In vivo heating of the guinea-pig fetal brain by pulsed ultrasound and estimates of thermal index. AB - Temperature was measured in the brain in live near-term fetal guinea pigs (62-66 d gestational age), during in utero exposure to a fixed beam of pulsed ultrasound at intensity ISPTA 2.82 W/cm2. Mean temperature increases of 4.3 degrees C close to parietal bone and 1.1 degrees C in the mid-brain were recorded after 2-min exposures. These values were lower (12%) than those obtained for ultrasound induced heating near the bone in dead fetuses insonated in utero. A significant cooling effect of vascular perfusion was observed only when guinea pig fetuses reached late gestation, near term, when the cerebral vessels were well developed. The estimated value for the thermal index (TIB), as used in AIUM/NEMA output display standard, underestimated the measured temperature increase at the bone brain interface. The ratio of measured temperature to the TIB is 1.3. A modification of the cranial thermal index provided a more reasonable, conservative, estimate of the temperature increase at a biologically significant point of interest at the brain-bone interface. PMID- 10385969 TI - Treatment of implanted liver tumors with focused ultrasound. AB - This article reports treatment of implanted liver tumors (HSN fibrosarcoma) with focused ultrasound (FUS). Experiments were carried out on implanted liver tumors in vivo. In order to determine the optimum treatment conditions, various combinations of exposure parameters were investigated. The results showed that it is possible to achieve total destruction of tumor cells in the treatment volume using an FUS system with a frequency of 1.7 MHz, with in situ ISAL of 261 W/cm2, 5-s exposure duration, and 1.5-mm exposure separation, with an in situ ISAL of 266 W/cm2, 10-s duration, and 2-mm separation, or with in situ ISAL of 213 W/cm2, 8-s duration, and 1.5-mm separation. Fifteen selected tumors were treated with these experimentally determined "optimum" exposure conditions. All the tumors were destroyed completely. Assessment of tumor viability in the treated volume was performed using both histologic and tissue culture methods. The mechanism of tumor damage, the limitations of the tumor model, and the effect of exposure parameters and liver blood flow on the treatment are discussed. PMID- 10385970 TI - A 3-D finite-element model for computation of temperature profiles and regions of thermal damage during focused ultrasound surgery exposures. AB - Although there have been numerous models implemented for modeling thermal diffusion effects during focused ultrasound surgery (FUS), most have limited themselves to representing simple situations for which analytical solutions and the use of cylindrical geometries sufficed. For modeling single lesion formation and the heating patterns from a single exposure, good results were achieved in comparison with experimental results for predicting lesion size, shape and location. However, these types of approaches are insufficient when considering the heating of multiple sites with FUS exposures when the time interval between exposures is short. In such cases, the heat dissipation patterns from initial exposures in the lesion array formation can play a significant role in the heating patterns for later exposures. Understanding the effects of adjacent lesion formation, such as this, requires a three-dimensional (3-D) representation of the bioheat equation. Thus, we have developed a 3-D finite-element representation for modeling the thermal diffusion effects during FUS exposures in clinically relevant tissue volumes. The strength of this approach over past methods is its ability to represent arbitrarily shaped 3-D situations. Initial simulations have allowed calculation of the temperature distribution as a function of time for adjacent FUS exposures in excised bovine liver, with the individually computed point temperatures comparing favorably with published measurements. In addition to modeling these temperature distributions, the model was implemented in conjunction with an algorithm for calculating the thermal dose as a way of predicting lesion shape. Although used extensively in conventional hyperthermia applications, this thermal dose criterion has only been applied in a limited number of simulations in FUS for comparison with experimental measurements. In this study, simulations were run for focal depths 2 and 3 cm below the surface of pig's liver, using multiple intensity levels and exposure times. The results also compare favorably to published in vitro experimental measurements, which bodes well for future application to more complex problems, such as the modeling of multiple lesion arrays within complex anatomical geometries. PMID- 10385971 TI - Ultrasound-induced temperature increase in guinea-pig fetal brain in utero: third trimester gestation. AB - Temperature increase was measured at various depths in the brain of living fetal guinea pigs during in utero exposure to unscanned pulsed ultrasound at ISPTA 2.8 W/cm2. Mean temperature increases of 4.9 degrees C close to parietal bone and 1.2 degrees C in the midbrain were recorded after 2-min exposures. When exposures were repeated on the same sites in each fetus after death, the corresponding mean temperature increases were 4.9 degrees C and 1.3 degrees C, respectively. Cerebral blood perfusion had little cooling effect on ultrasound-induced heating in the guinea pig fetus of 57-61 days gestational age. PMID- 10385972 TI - NMR study of a Lewis(X) pentasaccharide derivative: solution structure and interaction with cations. AB - The structure and conformation of the synthetic pentasaccharide Gal(beta 1 4){Fuc(alpha 1-3)}GlcNAc(beta 1-3)Gal(beta 1-4)Glc-beta OMe of the Lewis(X) family has been determined by NMR spectroscopy in dimethyl sulfoxide and methanol. In these solvents, the binding constants with calcium have been evaluated as 9.5 and 29.6 M-1, respectively. Study of the interaction sites has been achieved through the use of paramagnetic divalent cations and distance triangulation methods. Two regions have been found, the first one in the vicinity of the fucose unit, the second one closer to the lactose part. PMID- 10385974 TI - A highly convergent and effective synthesis of the phytoalexin elicitor hexasaccharide. AB - The peracetylated hexasaccharide 1,2,4-tri-O-acetyl-3-O-(2,3,4,6-tetra-O-acetyl beta-D-glucopyranosyl)-6- O- (2,3,4-tri-O-acetyl-6-O-(2,4-di-O-acetyl-3,6-di-O (2,3,4,6-tetra-O-acety l- beta-D-glucopyranosyl)-beta-D-glucopyranosyl)-beta-D glucopyranosyl)-alp ha, beta-D-glucopyranose 21 was synthesized in a blockwise manner, employing trisaccharide trichloroacetimidate 2,4-di-O-acetyl-3,6-di-O (2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl)- alpha-D-glucopyranosyl trichloroacetimidate 17 as the glycosyl donor, and trisaccharide 4-O-acetyl-3-O (2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl)-6-O-(2,3,4 -tri -O-acetyl-beta-D glucopyranosyl)-1,2-O-(R,S)ethylidene-alpha-D-glucopyra nose 18 as the acceptor. The donor 17 and acceptor 18 were readily prepared from trisaccharides 3-O (2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl)-6-O-(2,3,4-tri-O-acet yl- 6-O chloroacetyl-beta-D-glucopyranosyl)-1,2-O-(R,S)ethylidene-alpha-D- glucopyranose 10 and 3,6-di-O-(2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl)-1,2-O-(R,S) ethylidene-alpha-D-glucopyranose 11, respectively, which were obtained from rearrangement of orthoesters 3,4-di-O-acetyl-6-O-chloroacetyl-alpha-D glucopyranose 1,2-(3-O-(2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl)-1,2-O-(R,S) ethylidene-alpha-D-glucopyranosid-6-yl orthoacetate) 8 and 3,4,6-tri-O-acetyl alpha-D-glucopyranose 1,2-(3-O-(2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl)-1,2 O-(R,S) ethylidene-alpha-D-glucopyranosid-6-yl orthoacetate) 9, respectively. The orthoesters were prepared from selective coupling of the disaccharide 3-O (2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl)-1,2-O-(R,S) ethylidene-alpha-D glucopyranose 4 with 'acetobromoglucose' (tetra-O-acetyl-alpha-D-glucopyranosyl bromide) and 6-O-chloroacetylated 'acetobromoglucose', respectively. To confirm the selectivity of the orthoester formation and rearrangement, the disaccharide 4 O-acetyl-3-O-(2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl)-1,2-O-(R,S ) ethylidene-alpha-D-glucopyranose 7 was prepared from 4 by selective tritylation, acetylation and detritylation. The title compound, an elicitor-active D glucohexaose 3-O-(beta-D-glucopyranosyl)-6-O-(6-O-(3,6-di-O-(beta-D glucopyranosyl)-b eta -D-glucopyranosyl)-beta-D-glucopyranosyl)-alpha,beta-D glucopyranose 1, was finally obtained by Zemplen deacetylation of 21 in quantitative yield. PMID- 10385973 TI - Conformational analysis of an alpha-galactosyl trisaccharide epitope involved in hyperacute rejection upon xenotransplantation. AB - alpha-Galactosyl epitopes are carbohydrate structures bearing an alpha-Gal-(1- >3)-Gal terminus (alpha-Gal epitopes). The interaction of these epitopes on the surface of animal cells with anti alpha-Gal antibodies in human serum is believed to be the main cause in antibody-mediated hyperacute rejection in xenotransplantation. In this paper, conformational analysis of an N-linked alpha D-Galp-(1-->3)-beta-D-Galp-(1-->4)-beta-D-Glcp trisaccharide epitope was conducted in terms of each monosaccharide residue conformation, primary hydroxymethyl group configuration, and interglycosidic conformations. Selective 2D J-delta INEPT experiments have been carried out at three different temperatures to evaluate three-bond, long-range 13C-1H coupling constants for the crucial alpha-(1-->3) linkage. The NMR experimental data were complemented by theoretical calculations. The flexibility and dynamics of the trisaccharide have been studied by Metropolis Monte Carlo simulations. Ensemble-averaged three-bond, long-range 13C-1H coupling constants and nuclear Overhauser effects were in good agreement with the experimental data. The alpha-(1-->3) glycosidic linkage has shown a restricted flexibility as indicated by NMR spectroscopy and molecular modeling. PMID- 10385975 TI - A highly efficient and convergent synthesis of a hexasaccharide, a dimer of the repeating unit of the antigen O2 polysaccharide of Stenotrophomonas maltophilia. AB - A highly efficient and convergent synthesis of a hexasaccharide, which is a dimer of the repeating unit of the antigen O2 polysaccharide of Stenotrophomonas maltophilia, was achieved via coupling of 2,3,4-tri-O-acetyl-alpha-L xylopyranosyl bromide with the tetrasaccharide, allyl 4-O-{3-O-[4-O-(3,4-di-O benzoyl-alpha-L-rhamnopyranosyl)-2,3,6-tri-O-ben zoyl -alpha-D-mannopyranosyl]-4 benzoyl-alpha-L-rhamnopyranosyl}-2,3,6-tri-O- benzoyl-alpha-D-mannopyranoside (18) by the Koenigs-Knorr method followed by deacylation. Compound 18 was readily prepared from the coupling of the disaccharide trichloroacetimidate, 4-O-(2-O acetyl-3,4-di-O-benzoyl-alpha-L-rhamnopyranosyl)-2,3,6-tri-O- benzoyl-alpha-D mannopyranosyl trichloroacetimidate (8) with the disaccharide acceptor, allyl 4-O (2-O-acetyl-4-O-benzoyl-alpha-L-rhamnopyranosyl)-2,3,6-tri-O-benzoyl - alpha-D mannopyranoside (16), and both 8 and 16 were prepared via the trichloroacetimidate method from simple starting materials. The sole use of acyl protecting groups substantially simplified protection and deprotection, and the allyl group at the reducing end of allyl 4-O-{2-O-[2,3,4-tri-O-acetyl-beta-L xylopyranosyl]-3-O-[4-O-(2-O-(2,3,4- tri-O-acetyl-beta-L-xylopyranosyl)-3,4-di-O benzoyl-alpha-L-rhamnopyrano syl) -2,3,6-tri-O-benzoyl-alpha-D-mannopyranosyl]-4 O-benzoyl-alpha- L-rhamnopyranosyl}-2,3,6-tri-O-benzoyl-alpha-D-mannopyranoside 19 allowed further chemical transformation. PMID- 10385976 TI - Enzymatic synthesis of Kdn oligosaccharides by a bacterial alpha-(2-->6) sialyltransferase. AB - Synthesis of CMP-deaminoneuraminic acid (CMP-beta-D-Kdn) and its enzymatic transfer reaction using bacterial alpha-(2-->6)-sialyltransferase were examined. CMP-beta-D-Kdn was prepared from methyl 4,5,7,8,9-penta-O-acetyl-3-deoxy-D glycero-beta-D-galacto-2- nonulopyranosonate (2) in 24% overall yield. Enzymatic synthesis of Kdn oligosaccharide with CMP-beta-D-Kdn (10.2 mumol), methyl beta-D lactosaminide (7, 8.1 mumol) and purified sialyltransferase (80 munits) afforded Kdn-alpha-(2-->6)-Gal-beta-(1-->4)-GlcNAc-beta-1-OMe in 77% yield. PMID- 10385977 TI - Isolation and characterization of new limonoid glycosides from Citrus unshiu peels. AB - Three limonoid glycosides were isolated from Citrus unshiu peels, and their structures were determined based on MS and NMR spectroscopic data as nomilinic acid 17-O-beta-D-glucopyranoside (1), methyl nomilinate 17-O-beta-D glucopyranoside (2), and obacunone 17-O-beta-D-glucopyranoside (3). In particular, the location of the sugar moiety was clearly determined by the B/E constant linked scan FABMS method. No limonoid glycosides obtained here were found to have antitumor activity in NCI-H292 and EL-4 cell lines. PMID- 10385979 TI - Structural determination of the acidic exopolysaccharide produced by a Pseudomonas sp. strain 1.15. AB - Pseudomonas strain 1.15 was isolated from a freshwater biofilm and shown to produce considerable amounts of an acidic polysaccharide which was investigated by methylation analysis, NMR spectroscopy and ionspray mass spectrometry (ISMS). The polysaccharide was depolymerised by a bacteriophage-associated endoglucosidase and by autohydrolysis, and the resulting oligosaccharides were investigated by NMR spectroscopy and mass spectrometry. The resulting data showed that the parent repeating unit of the 1.15 exopolysaccharide (EPS) is a branched hexasaccharide. The main chain is constituted of the trisaccharide -->4)-alpha-L Fucp-(1-->4)-alpha-L-Fucp-(1-->3)-beta-D-Glcp- (1--> and the side chain alpha-D Galp-(1-->4)-beta-D-GlcAp-(1-->3)-alpha-D-Galp-(1-->is linked to O-3 of the first Fuc residue. The terminal non-reducing Gal carries a 1-carboxyethylidene acetal in the R configuration at the positions 4 and 6. Of the four different O-acetyl groups present in non-stoichiometric amounts, two were established to be on O-2 of the 3-linked Gal and on O-2 of the 4-linked Fuc. PMID- 10385978 TI - A simple synthesis of 8-(methoxycarbonyl)octyl 3,6-di-O-(alpha-D-mannopyranosyl) alpha-D-mannopyranoside and derivatives and their use in the preparation of neoglycoconjugates. AB - 8-(Methoxycarbonyl)octyl 3,6-di-O-(alpha-D-mannopyranosyl)-alpha-D mannopyranoside (10) was synthesized in 54% yield by regioselective diglycosylation of unprotected mannoside 4, employing the trichloroacetimidate donor 1, followed by debenzoylation. Derivatives of compounds 4 and 10 were used to prepare conjugates containing fluorochromes for the study of carbohydrate lectin interactions, as well as conjugates with phospholipids for the preparation of liposomes. PMID- 10385980 TI - Processing linguistic and perceptual dimensions of speech: interactions in speeded classification. AB - Performance on selective-attention and divided-attention tasks shows strong and consistent interactions when participants rapidly classify auditory stimuli whose linguistic and perceptual dimensions (the words low vs. high, low and high pitch, low and high position in space) share common labels. Compared with baseline performance, response times were greater when one or two irrelevant dimensions varied (Garner interference) and when combinations of attributes were incongruent rather than congruent (congruence effects). Performance depended only on the congruence relationships between the relevant dimension and each of the irrelevant dimensions and not on the congruence relationships between the irrelevant dimensions themselves. In selective attention, an additive multidimensional model accounts well for the patterns of both Garner interference and congruence effects. PMID- 10385981 TI - Auditory objects of attention: the role of interaural time differences. AB - The role of interaural time difference (ITD) in perceptual grouping and selective attention was explored in 3 experiments. Experiment 1 showed that listeners can use small differences in ITD between 2 sentences to say which of 2 short, constant target words was part of the attended sentence, in the absence of talker or fundamental frequency differences. Experiments 2 and 3 showed that listeners do not explicitly track components that share a common ITD. Their inability to segregate a harmonic from a target vowel by a difference in ITD was not substantially changed by the vowel being placed in a sentence context, where the sentence shared the same ITD as the rest of the vowel. The results indicate that in following a particular auditory sound source over time, listeners attend to perceived auditory objects at particular azimuthal positions rather than attend explicitly to those frequency components that share a common ITD. PMID- 10385982 TI - Development and phonetic differentiation of speech movement patterns. AB - It is often hypothesized that speech production units are less distinctive in young children and that generalized movement primitives, or templates, serve as a base on which distinctive, mature templates are later elaborated. This hypothesis was examined by analyzing the shape and stability of single close-open speech movements of the lower lip recorded in 4-year-old, 7-year-old, and adult speakers during production of utterances that varied in only a single phoneme. To assess the presence of a generalized template, lower lip movement sequences were time and amplitude normalized, and a pattern recognition procedure was implemented. The findings indicate that speech movements of children already converged on phonetically distinctive patterns by 4 years of age. In contrast, an index of spatiotemporal stability demonstrated that the stability of underlying patterning of the movement sequence improves with maturation. PMID- 10385983 TI - On the distinction between visual salience and stimulus-driven attentional capture. AB - It is often assumed that the efficient detection of salient visual objects in search reflects stimulus-driven attentional capture. Evidence for this assumption, however, comes from tasks in which the salient object is task relevant and therefore may elicit a deliberate deployment of attention. In 9 experiments, participants searched for a nonsalient target (vertical among tilted bars). In each display, 1 bar was highly salient in a different dimension (e.g., color or motion). When the target and salient elements coincided only rarely, reducing the incentive to attend deliberately to the salient stimuli, response times depended little on whether the target was salient, although some interesting exceptions were observed. It is concluded that efficient selection of an element in visual search does not constitute evidence that the element captures attention in a purely stimulus-driven fashion. PMID- 10385984 TI - Sequential priming in hierarchically organized figures: effects of target level and target resolution. AB - Three experiments are reported in which participants identified target letters that appeared at either the global or local level of hierarchically organized stimuli. It has been previously reported that response time is facilitated when targets on successive trials appear at the same level (L. M. Ward, 1982; L. C. Robertson, 1996). Experiments 1 and 2 showed that this sequential priming effect can be mediated by target-level information alone, independent of the resolution, or actual physical size, of targets. Target level and resolution were unconfounded by manipulating total stimulus size, such that global elements of the smaller stimuli subtended the same amount of visual angle as local elements of the larger stimuli. Experiment 3, however, showed that when level information is less useful than resolution in parsing targets from distractors, resolution does become critical in intertrial priming. These data are discussed as they relate to the role of attention in local vs. global (part vs. whole) processing. PMID- 10385985 TI - Manual laterality and hitting performance in major league baseball. AB - Asymmetrical hand function was examined in the context of expert sports performance: hitting in professional baseball. An archival study was conducted to examine the batting performance of all Major League Baseball players from 1871 to 1992, focusing on those who batted left (n = 1,059) to neutralize the game asymmetry. Among them, left-handers (n = 421) were more likely to hit with power and to strike out than right-handers (n = 638). One possible account, based on the idea of hand dominance and an analogy to tennis, is that batting left involves a double-handed forehand for left-handers and a weaker and more reliable double-handed backhand for right-handers. The results are also interpretable in the light of Y. Guiard's (1987) kinematic chain model of a between-hands asymmetrical division of labor, which provides a detailed account of why left batting is optimal for left-handers. PMID- 10385986 TI - Manipulation with no or partial vision. AB - The present research investigated the role of vision in closed- and open-loop processing during manipulation. In Experiment 1, participants performed common manipulatory tasks with 100% accuracy in less than 1 s without vision. In Experiment 2, the effects of extensive practice of a peg-in-hole task were examined within 4 functionally significant stages of manipulation. Performance was consistently faster with than without vision in the prereach, grasp, and transport + insert stages; reverse effects were observed during the reach stage. In Experiment 3, the effects of practice with partial vision were examined: Participants initially learned the peg-in-hole task with full vision and then transferred to learning the same task with vision available only during 1 functional stage. Overall, performance was fastest when vision was limited to the prereach and reach stages. PMID- 10385987 TI - Identity priming in English is compromised by phonological ambiguity. AB - If it takes longer to achieve a single phonological representation for inconsistent words (e.g., BOWL) than for consistent words (e.g., BENT), and if phonological coherence is pivotal to visual word recognition, then identity priming should depend on consistency. This hypothesis was evaluated in naming and lexical decision within a 4-field presentation sequence of mask-prime-mask target. The prime-target stimulus onset asynchrony (SOA) was either 114 or 244 ms (with prime durations, respectively, of 43 and 129 ms). Four experiments compared identity primes such as BOWL and BENT, which were equated, on average, for total number of friendly and unfriendly neighbors, bigram frequency, and number of 1 letter-different neighbors. In both tasks, BENT primed itself better than BOWL primed itself, with the difference being larger at the shorter SOA. Word processing is constrained primarily by the rate of achieving a coherent phonological code. PMID- 10385988 TI - Perceptual-motor sequence learning of general regularities and specific sequences. AB - Participants viewed digit strings and typed them on a computer keyboard. When they used the same key configuration across training and test, they typed test strings that adhered to the same sequence rule as training strings faster than test strings that adhered to the opposite rule (general-regularity [GR] learning), and they typed test strings that were processed repeatedly during training faster than test strings that were not (specific-sequence [SS] learning). However, when they used different key configurations at training and at test, GR learning, but not SS learning, was observed. Conversely, when they did not type but spoke the strings aloud during training, SS learning, but not GR learning, was observed. Results suggest that in addition to declarative memory for specific sequences, relatively independent subsystems underlie procedural learning of perceptual-motor sequence components (producing GR effects) and sequence wholes (producing SS effects). PMID- 10385989 TI - Noise, information transmission, and force variability. AB - This study was designed to test the hypothesis derived from information theory that increases in the variability of motor responses result from increases in perceptual-motor noise. Young adults maintained isometric force for extended periods at different levels of their maximum voluntary contraction. Force variability (SD) increased exponentially as a function of force level. However, the signal-to-noise ratio (M/SD), an index of information transmission, as well as measures of noise in both the time (approximate entropy) and frequency (power spectrum) domains, changed according to an inverted U-shaped function over the range of force levels. These findings indicate that force variability is not directly related to noise but that force output noisiness is positively correlated with the amount of information transmitted. PMID- 10385990 TI - Haptic perception of the distance walked when blindfolded. AB - The ability to detect the distance walked when blindfolded using only haptic information generated by the walking activity was investigated. Participants walked a straight path until told to stop, turned, and attempted to return to their starting point. The path was completely featureless. Counting was prevented. Blindfolded, sighted participants traveled with a long cane or a trained sighted guide. Gait was varied or distorted. In all experiments the return distance was a linear function of the set distance, with some participants giving and some conditions resulting in remarkably sensitive performances. The magnitude of errors was a linear function of step length. PMID- 10385991 TI - Customisation of AFLP analysis for cassava varietal identification. AB - Amplified fragment length polymorphism (AFLP) markers were used in the characterization of eight cassava varieties. This nonradioactive AFLP system was customized in terms of the choice of restriction enzymes used and the selection of nucleotides added to the 3' end of primers. EcoRI/MseI and HindIII/MseI fragments generally gave monomorphic profiles while ApaI/TaqI fragments produced polymorphic profiles suggesting a genome with high G + C content. It was possible to identify the eight cassava varieties used in this study using CTG as selective bases at the TaqI primer. For cassava, the AFLP system provided a higher number of loci detected per run when compared to RAPD. The reliability accompanying AFLP analysis would thus make it suitable for the characterization of cassava varieties. PMID- 10385992 TI - Cytochrome P450 monooxygenases of DIBOA biosynthesis: specificity and conservation among grasses. AB - DIBOA and DIMBOA are secondary metabolites of grasses which function as natural pesticides. The four maize genes BX2 through BX5 encode cytochrome P450-dependent monooxygenases that catalyse four consecutive reactions in the biosynthesis of these secondary products. Although BX2-BX5 share significant sequence homology, the four enzymes have evolved into specific enzymes each catalysing predominantly only one reaction in the pathway. In addition to these natural reactions, BX3 hydroxylates 1,4-benzoxazin-3-one and BX2 shows pCMA demethylase activity. With respect to DIBOA biosynthesis, identical enzymatic reactions have been found in rye as compared to maize, indicating early evolution of the P450 enzymes in the grasses. PMID- 10385993 TI - Proteolytic activity in relation to seasonal cambial growth and xylogenesis in Pinus banksiana. AB - Proteolytic activity in the cambial zone and developing xylem of Pinus banksiana Lamb. was investigated over an annual cycle of growth and dormancy. Highest proteolytic activity was associated with the most active period of primary-wall radial expansion of cambial derivatives, in early spring, before protoplasmic autolysis was initiated in developing earlywood. Three pH maxima of proteolytic activity, near pH 3.0, 6.5 and 9.5, were observed at that time. In general, activities measured at pH values below 7.0 were greater than those determined above pH 7.0 at all stages in the annual cycle, in both cambial zone and developing xylem, although elevated activity at alkaline pH was also observed during springtime growth. Polyvinylpolypyrollidone (PVP) treatment markedly enhanced pH 7.5 but not pH 4.0 proteolytic activity in the cambial zone, but not in developing xylem, indicating the presence of PVP-binding proteinase regulators in the cambium. By fractionation and effector studies total proteolysis was determined to comprise interactions between serine, cystine, aspartate and metallo-proteases having MWs, by gel chromatography, between 10 and 100 kDa. The observations point to a complex regulatory mechanism controlling the presence and catalytic rates of the distinct types of proteases in the cambial region throughout an annual cycle of growth and dormancy. PMID- 10385994 TI - A novel elicitin necrotic site revealed by alpha-cinnamomin sequence and site directed mutagenesis. AB - Elicitins are 10 kDa proteins secreted by Phytophthora fungi, that elicit resistance against certain plant pathogens. Various natural molecules, mutated recombinant elicitins and synthetic peptides were previously shown to differentially induce in tobacco leaf necrosis and defence genes, activities borne by several sites which were identified. We report a novel necrosis determining residue at position 25, revealed by the comparison of the necrotic activity and sequence of alpha-cinnamomin with those of other known elicitins. Using a modified recombinant beta-cryptogein, expressed in Pichia pastoris, we show that the substitution of asparagine 25 by a serine leads to a significant enhancement of the necrotic activity. PMID- 10385995 TI - Synthesis and evaluation of 4-(3-methyl-2-butenoxy) isonitrosoacetophenone, a radiation-induced stress metabolite in Citrus. AB - The time- and dose-dependent occurrence of 4-(3-methyl-2 butenoxy)isonitrosoacetophenone, a gamma-irradiation-induced stress metabolite was investigated. The chemical synthesis of the compound is reported. The compound exhibits antifungal activity, as well as antioxidant activity, as indicated by its ability to scavenge reactive oxygen radicals in a chemiluminescence assay. PMID- 10385997 TI - Immunological and molecular comparison of polyphenol oxidase in Rosaceae fruit trees. AB - An antibody raised against apple polyphenol oxidase (PPO) cross-reacted with PPOs from Japanese pear (Pyrus pyrifolia), pear (Pyrus communis), peach (Prunus persica), Chinese quince (Pseudocydonia sinensis) and Japanese loquat (Eriobotrya japonica). Core fragments (681 bp) of the corresponding PPO genes were amplified and characterized. The deduced protein sequences showed identities of 85.3 to 97.5%. Chlorogenic acid oxidase activity of these PPOs showed higher activities when assayed at pH 4 than at pH 6. These results indicate that PPOs in Rosaceae plants are structurally and enzymatically similar. PMID- 10385998 TI - Asitrilobins A and B: cytotoxic mono-THF annonaceous acetogenins from the seeds of Asimina triloba. AB - The seeds of Asimina triloba have yielded two novel cytotoxic mono tetrahydrofuran (THF) Annonaceous acetogenins, asitrilobins A (1) and B (2). In addition, annonacin, asimin and asiminacin, which are known, and annomontacin and xylomaticin, which are known but are new in this species, were obtained. Compounds 1 and 2 have a relative stereochemical relationship of erythro/cis/threo across the mono-THF ring with its two flanking hydroxyls and they, thus, represent a new type of acetogenin. Their structures were established on the basis of chemical and spectral evidence. 1 and 2 showed potent bioactivities in the brine shrimp lethality test (BST) and among six human solid tumor cell lines with notable selectivity for the pancreatic cell line (MIA PaCa 2) at ten to one-hundred times the potency of adriamycin. PMID- 10385999 TI - Growth factors and their receptors in the olfactory system. AB - The olfactory epithelium is unique in the mammalian nervous system as it is a site of continual neurogenesis. Constant turnover of primary sensory neurons in the periphery results in continuous remodeling of neuronal circuits and synapses in the olfactory bulb throughout life. Most of the specific mechanisms and factors that control and modulate this process are not known. Recent studies suggest that growth factors, and their receptors, may play a crucial role in the development and continuous regeneration of olfactory neurons, i.e. particularly in neuronal proliferation, neurite outgrowth, fasciculation and synapse formation of the olfactory system. The potential role of the following factors and their receptors in different species are reviewed: Nerve growth factor (NGF); insulin like growth factors (IGFs); fibroblast growth factors (FGFs); epidermal growth factor (EGF); transforming growth factor alpha (TGF alpha); amphiregulin (AR) and transforming growth factors beta (TGFs beta). PMID- 10386000 TI - An immunohistochemical survey of catecholamine-synthesizing enzyme-immunoreactive nerves and endocrine cells in the bovine pancreas. AB - The distribution of catecholamine-synthesizing enzyme-immunoreactive nerves and endocrine cells in the pancreas of the calf and cow was studied immunohistochemically using antisera against tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH). TH- and DBH-immunoreactive nerve fibres were abundant both within and around the islet of Langerhans. A few TH- and DBH immunoreactive nerve fibres were seen around the large islets characteristic of calf pancreas, but the majority of cells in the large islets, and some in islets of Langerhans, showed TH immunoreactivity. In the exocrine pancreas, both TH- and DBH-immunoreactive nerve fibres were distributed randomly among the acini, with the DBH-immunoreactive fibres being more numerous. Abundant TH- and DBH immunoreactive nerve fibres were seen in close association with blood vessels and in the connective tissue around the interlobular duct. Immunoreactivity for both enzymes was also observed in the nerve cell bodies and fibres of the intrapancreatic ganglia. The findings suggest an important role for catecholamines in the regulation of bovine pancreatic function. PMID- 10386001 TI - Morphological study of mast cell localization in the wall of the proximal tubule in the domestic swine kidney. AB - Morphological studies to localize mast cells in the wall of the convoluted and straight parts of the proximal tubules of the domestic swine kidney were performed. Kidney perfusion was carried out in 8-month-old pigs and semi-thin and ultrathin sections were prepared after a routine treatment. A light microscopic study showed that mast cells were not frequently found. Most were localized in the wall of the proximal convoluted tubule, while in the proximal straight tubule wall they were fewer in number. Most of the mast cells were oval in shape, their size being typical for this species, and the granules were reddish in colour. Three types of granule containing an evenly distributed granular-fibrous substance were found by TEM examination: the first type had a single electron optically dense belt under the membrane; the second showed optically empty spaces between the substance and the cell membrane; the granules from the third type had two parallel dense belts below the granular membrane and were the smallest in size. Some of the granules of the first type were connected with the granular endoplasmic reticulum. A very narrow intercellular area and single desmosome-like junctions were found between the mast cells and the epithelial cells. Protrusions of plasmalemma and granules outside the cells were not established. On the basis of these results and data from the scientific literature, a comparative analysis was carried out. This showed certain differences between the results obtained and previous findings on the morphology of domestic swine mast cells and granules. PMID- 10386002 TI - Anatomicohistological characteristics of the ovary of the coypu (Myocastor coypus). AB - Using histological, histochemical and macroscopic and microscopic measurement techniques, the macroscopic and microscopic structures of coypu ovaries were studied in sexually mature virgin females. The mature ovaries of the coypu were ovoid or elongated bodies, not encapsulated, covered by a single layer of epithelium. They had a parenchyma formed by follicles at different stages of evolution and true and accessory corpora lutea. The interstitial tissue was a prominent and permanent structure in the ovaries. Some ovaries contained a few rete ovarii in the hilus. PMID- 10386003 TI - Macroscopic cryosectioning: a simple new method for producing digital, three dimensional databases in veterinary anatomy. AB - Using a new method derived from the 'visible human project' (Spitzer et al., 1996, Journal of the American Medical Informatics Association, 3, 118-130), we were able to establish a simple and low-cost tool which produces high-quality cryosections of macroscopic specimens down to 1-mm slice thickness, based on a milling process. For the first time, a macroscopic cryotome is available to veterinary anatomists, which can be used on cutting faces up to 25 cm high and 50 cm wide and with a minimal slice thickness of 1 mm without any gap. The method employs a modified wood circular saw. Recording of the cutting faces is carried out 'online' by a high-resolution digital camera. The process has been tested extensively and produces high-quality sections of very hard material (teeth) as well as of very soft tissues (brain). It is now possible in veterinary medicine to provide three-dimensional anatomical databases of high resolution and of tissue-specific colour as an additional tool for high-quality two- and three dimensional anatomical reconstructions for use in science and education. PMID- 10386004 TI - How structures in bovine hoof epidermis are influenced by nutritional factors. AB - The structure of the hoof epidermis is the link between nutrition and horn quality. The aim of this study was to demonstrate the relationship of single structures in the process of keratinization and cornification of bovine hoof epidermis to certain nutritional factors such as lipids, minerals and vitamins. Furthermore, we wanted to show the structural changes in the dyskeratotic epidermis caused by an insufficient supply of keratinizing epidermal cells. For our study we used samples of hoof epidermis from 25 dual-purpose dairy cattle, with ages ranging between 2.5 and 4 years. We also obtained a complete set of hooves from a biotin-deficient calf. All samples were investigated by light and transmission electron microscopy, using routine methods as well as histochemical and enzyme-histochemical techniques. We focused on epidermal structures that have a major influence on horn quality and are known to be related to single nutritional factors. The strength of the keratin filament bundles is determined by their cross-linking via sulphur-containing amino acids. Essential fatty acids are required for the synthesis of an intercellular cementing substance connecting the horn cells and establishing a permeability barrier in the stratum corneum. Minerals, in particular calcium, are essential for activation of enzymes that are a prerequisite for physiological keratinization and cornification. Furthermore, vitamins such as biotin are essential in the metabolism of the keratinizing epidermal cells. PMID- 10386005 TI - Further observations on the presence of ganglion cells in the oculomotor nerves of mammals and fish: number, origin and probable functions. AB - The oculomotor nerves (3rd, 4th and 6th) of some species of fish and mammals have been studied to establish the presence, number, true topography and probable functional role of the ganglion cells located along the trunk. The finding of typical pseudo-unipolar ganglion cells is always unpredictable and extremely variable, from an inter- and intra-specific point of view, in members of the two zoological classes studied. PMID- 10386006 TI - [Evolution and Darwinian theory]. AB - The Darwinian theory today is broadly accepted as one cornerstone of science. However, isn't biology as a life science both, material- and art subject? PMID- 10386007 TI - Expression of angiotensinogen in the uterus induced by coagulating gland renin in mice. AB - Coagulating gland (CG) renin is one of the components of the local renin angiotensin system (RAS), which plays important roles in the maintenance of homeostasis in several tissues. Although the existence of local renin has been reported in many tissues, its function is not yet well understood. In the present study, the relationship between CG renin and uterine angiotensinogen was investigated by immunohistochemical and in situ hybridization techniques. In mating experiments with male and female C57BL/6 mice, renin was demonstrated immunohistochemically on the epithelial cells of the uterus on the first day after coitus; however, it was not detected on the second and third days after coitus. Neither immunoreactive cells nor messenger signals for renin were demonstrated on the epithelial cells of the uterus throughout the experiments. On the first day after mating, it was noted that positive signals for angiotensinogen mRNA were expressed on the epithelial cells of the uterus in situ hybridization, but not on the second and third days after coitus. These results suggest the possibility that CG renin is transferred to the uterine epithelium at mating and temporarily activates the expression of angiotensinogen in the uterus. Angiotensin II produced by angiotensinogen may act as an important mediator for vascularization of the first step of pregnancy. PMID- 10386008 TI - Immunohistochemical study of steroidogenic enzymes in the ovary and placenta during pregnancy in the dog. AB - Using the immunohistochemical technique, we attempted to identify the source of secretion of steroid hormones between the mid- and late-terms of gestation in dogs by investigating steroid converting enzymes such as cholesterol side-chain cleavage enzyme (SCC), 3 beta-hydroxysteroid dehydrogenase/isomerase (3 beta HSD), 17 alpha-hydroxylase/C17, 20lyase (c17), and aromatase in the ovaries and placenta. Aromatase positive cells were slightly confirmed in luteal cells in the mid-term of gestation (day 40), whereas, in the late-stage (day 50 and 60), the number of aromatase positive cells had increased. However, the oestrogen precursor (c-17 positive cells), could barely be identified in the marginal regions of the corpora lutea (CL) and completely disappeared in the late-stage of gestation. The androgen precursors, convertase SCC and 3 beta-HSD, were confirmed in all regions of the CL during the mid-stage of gestation (day 40), showing particularly strong cell reactions in the marginal region of the CL. Yet, these positive reactions of SCC and 3 beta-HSD in the marginal region of the CL disappeared in the late-stage of gestation. Moreover, it was discovered that the number of SCC and 3 beta-HSD positive cells had decreased in all regions of the CL. None of the enzymes were detected in the placenta. The above results indicated that the source of oestrogen secretion in pregnant dogs is considered to be the CL, and that, compared with the mid-stage of gestation, there was an increased number of oestrogen synthesizing cells within the CL in the late-stage. However, the biosynthetic site of oestrogen precursors from the luteal cells during the late-stage of gestation is still unknown. PMID- 10386010 TI - [Comparative histological study of the female reproductive system in the llama (Lama guanicoe glama). II. Oviduct, uterus, cervix, vagina]. AB - The histological characteristics of the oviduct, uterus, cervix and vagina of the llama are described and compared with those of domestic mammals. There are differences between llama and domestic mammals in the type of placentation and the epithelium of the vagina, which seems to be a transitional epithelium. PMID- 10386009 TI - [Pulmonary blood vessels in goats]. AB - The blood vessels in the lung of the goat, which until now have received little attention, are described in detail for the first time. With regard to the segments of the lung, blood vessels are bronchovascular units in the lobi craniales, lobus medius and lobus accessorius, but bronchoartery units in the lobi caudales. We investigated the types of branches of the Aa. pulmonales dextra et sinistra, the inter- and intraspecific principles of the outlet of the pulmonary veins and the importance of bronchopulmonary segmentation of the lungs. PMID- 10386011 TI - Isolation and purification of penicillin G acylase obtained from Escherichia coli (NCIM-2400) and immobilisation on Eupergit C for the production of 6 amino penicillanic acid. AB - Penicillin G-Acylase is produced by submerged cultivation of E. Coli (NCIM-2400) and extracted from the harvested fermented broth, purified (affinity chromatography) and immobilised on Eupergit C (Synthetic polymer in bead form). The immobilised penicillin G acylase properties are studied and compared with soluble penicillin G-acylase. The control parameters for conversion of penicillin G-K to 6 APA are optimised [e.g. substrate (Pen G-K) concentration ratio to immobilised penicillin G-acylase, temperature, pH etc.] in a stirred tank reactor. Our findings suggest that immobilised penicillin G-acylase can be used commercially and the productivity of 1 kg. of immobilised enzyme is around 400 kg of 6 APA under given desired stipulated conditions. PMID- 10386012 TI - Auxotrophic requirement for sporangiospore--yeast transformation of Dimorphomyces diastaticus strain C 12. AB - The filamentous microorganism tentatively known as Dimorphomyces diastaticus originally isolated from fermenting soursop extract, in contradistinction grew in the considiogeneous form in glucose-ammonium sulphate-basal salts buffered medium which induced polar budding yeast cells of another dimorphic microorganism, D. pleomorphis (1). Further experiments using this medium showed that a number of growth factors including inositol, thymine and uracil separately incorporated inducted yeast morphology. Carbon substrates gave rise to polar budding yeast cells in the following order: galactose, malic acid, maltose, but mycelial fragments were preponderant in the reverse order. There was no growth with citric acid as substrate. With maltose-substrate and uracil incorporation in buffered basal salts medium, only polar budding yeast cells were induced, while granular particles abound with thymine supplementation. It was concluded that D. diastaticus strain C12 auxotrophically required a dissacharide, maltose, and pyrimidine base, uracil, for sporangiospore-yeast transformation in buffered ammonium sulphate-basal salts medium, pH 3.5. PMID- 10386013 TI - Affinity chromatography of maltase on aliphatic amines as ligands. AB - The primary amino group, the competitive inhibitor of maltase, was used as ligand and the enzyme was purified to homogeneity in a single step. The amino group affiants of ethylene (C2-NH2) and hexamethylene (C6-NH2) diamines were prepared by coupling to cyanogen bromide activated Sepharose CL-4B. The enzyme was quantitatively adsorbed at alkaline pH (pH 8.2), while the elution could be effected only in presence of maltose at acidic pH. The elution of enzyme by maltose was independent of spacer arms (C2 and C6) which suggests specific binding of the enzyme through inhibitor site. PMID- 10386014 TI - Microbial production of L-lysine: a review. AB - Microbial production of L-lysine has been reviewed with 251 references. The review includes different lysine producing microorganisms, their optimum cultural conditions, yield, assay and process of product recovery. It includes a discussion on the pathway of aspartate family of amino acid's biosynthesis in bacteria and its regulation. Achievements in this regard made through genetic engineering have also been included. PMID- 10386015 TI - A new variety of Chainia olivacea from marine sediment off Gulf of Mannar. AB - A new variety of Chainia olivacea was isolated from marine sediment off Gulf of Mannar. The morphological, cultural, physiological and biochemical characters were studied, compared to known species and identified as a new variety of Chainia olivacea. Antibiotic activity of the strain was tested against both Gram positive and Gram negative bacteria as well as fungi and yeasts. Sodium chloride tolerance was also tested. PMID- 10386016 TI - In vitro evaluation of inhibitory nature of extracts of 18-plant species of Chhindwara against 3-keratinophilic fungi. AB - Effect of extract of 18 plant species, viz., Acorus calamus, Adhatoda vasica, Amomum subulatum, Andrographis paniculata, Boerhaavia diffusa, Cassia occidentalis, Centella asiatica, Cymbopogon citratus, Hemidesmus indicus, Hyptis suaveolens, Malvestrum sp., Passiflora edulis, Pergularia daemia, Peristrophe bicalyculata, Shuteria hirsuta, Solanum nigrum, Tecoma stans, and Verbascum chinense on the growth of Microsporum gypseum, Chrysosporium tropicum and Trichophyton terrestre was evaluated and discussed. The sensitivity of the keratinophilic fungi was evaluated by dry-weight method. The maximum inhibition of mycelial growth was shown by M. gypseum (86.62%) followed by T. terrestre (81.86%) and C. tropicum (74.06%) when treated with S. hirsuta whereas the minimum inhibition was exhibited by M. gypseum (0.29%), C. tropicum (0.16%) and T. terrestre (1.76%) when tested with the extract of P. edulis, A. vasica and B. diffusa respectively. PMID- 10386017 TI - Antimycotic effects of some antibiotics on the growth of some dermatophytes and other keratin degrading fungi. AB - Four antibiotics have been tested against the growth of some dermatophytes and keratin degrading fungi. A gradual decrease in growth was observed with increase in concentration of all antibiotics. All, but griseofulvin observed to inhibit > 50% mycelial weight even at a lower concentration of 50 ppm. Azole derivatives were most toxic to the growth of M. gypseum at all concentrations, whereas, to that of C. tropicum at above 100 ppm. Mycostatin was the most toxic antibiotic to the growth of M. gypsea at all concentrations. PMID- 10386018 TI - Optimisation of cultural conditions for antifungal antibiotic accumulation by Streptomyces rochei G164. AB - To induce higher amount of antifungal antibiotic production by variation of cultural parameters has been studied. The maximum effectivity was found in sucrose as carbon source, peptone as nitrogen source and at pH 7.0. The effect of other selected factors were also evaluated in order to judge the variables that influenced antibiotic production. PMID- 10386019 TI - [Characterization of enterohemorrhagic Escherichia coli O26 and development of its isolation media]. AB - We studied 101 strains of Enterohemorrhagic Escherichia coli (EHEC) O26 isolated from diarrhea patients in six prefectural institutes of public health in Japan during June 1996 and December 1997 and tried to establish an isolation medium for EHEC O26. None of the 101 EHEC O26 strains fermented rhamnose; Whereas all of the other EHEC including O157 and non-EHEC (166 strains) fermented rhamnose except 1 strain of non-EHEC. All of the randomly selected EHEC O26 (14 strains of O26:H11.2 strains of O26:H-) showed a very high resistance to potassium tellurite (Minimal Inhibitory Concentration (MIC) > or = 50 micrograms/ml), whereas all of the randomly selected non-EHEC (26 strains) but 1 showed a high sensitivity (MIC < or = 6.25 micrograms/ml) to this compound. On the basis of these results, we developed a Rhamnose MacConkey (RMAC) medium in which lactose in the MacConkey medium was replaced by rhamnose, and Cefixime-Potassium Tellurite-RMAC (CT-RMAC) medium in which Cefixime (0.05mg/l) and Potassium Tellurite (25mg/l) was added to RMAC for the isolation of EHEC O26 strains. We then evaluated the specifcity of these selective media by growing a selected number of O26 (24 strains) and 9 selected strains of bacteria. All of the EHEC O26 strains generated rhamnose non fermented colonies (white color) on both media. In contrast to the EHEC O26, the vast majority of E. coli strains (166/167 = 99.4%) other than EHEC O26 were theoretically assumed to generate red colonies on the RMAC medium because of their rhamnose fermenting character and most of them were assumed not to grow on CT-RMAC medium because of their sensitivity to potassium tellurite. These findings and results indicate that EHEC O26 can be easily distinguished from other strains of E. coli including O157. Although EHEC O26 strains showed somewhat poor growth on CT-RMAC medium compared with that on RMAC medium, these O26 showed almost the same degree of growth on CT-RMAC as they showed on DHL media. The results of the present study demonstrated that the use of RMAC and CT MRAC media for the isolation of EHEC O26 is very reliable and efficient with RMAC having good sensitivity and CT-RMAC having a better specificity for the isolation of this strain of EHEC. PMID- 10386020 TI - [Strains of Shigella sonnei recently isolated in Tokyo]. AB - A total of 341 Shigella sonnei strains consisting of 94 domestic strains and 247 imported strains isolated during 1990-1997 in Tokyo, were examined regarding their colicine-type, drug-resistance and ornithine-utilization. The colicine typing results showed that the domestic strains were classified into 7 types, and the imported strains were classified into 13 types. Among the colicine-types identified, 8-type, 0-type, 6-type and 12-type were predominant in the domestic strains, whereas 6-type, 0-type, 8-type, 9A-type and 12-type were predominant in the imported strains. The drug-resistance test using 9 drugs (CP, TC, SM, KM, ABPC, ST, NA, FOM and NFLX) showed that 89.4% of the domestic strains and 85.4% of the imported strains were resistant to some of the drugs except FOM and NFLX. Drugs with a high resistant rate were SM, TC and ST for both groups. Drug resistance patterns of the resistant strains varied in 22 types. Among those, a triple drug-resistance type with TC.SM.ST was found in the most frequent pattern in both groups. The results of the ornithine-utilization test revealed that 28.7% of the domestic strains and 8.1% of the imported strains were negative. The ornithine-negative strains in the same source had a similar plasmid-profile, but generally there was no correlation between the different sources. PMID- 10386021 TI - [Evaluation of urinary antigen detection methods for rapid diagnosis of Legionella pneumonia]. AB - We have evaluated urine specimens of presumptive cases of legionnaires' disease (110 cases, 173 sample), collected in the past eight years (April, 1990-August, 1998) with the Binax EIA kit which detects the soluble antigen of Legionella pneumophila serogroup (SG) 1, and the Biotest EIA kit which detects Legionella species. Seven cases (19 specimens) were positive for the Binax EIA kit, and nine cases (22 specimens) were positive for the Biotest EIA kit. The sensitivity for culture, PCR, IFA method were 100%, 100%, and 50%, the specificity for these method were 93%, 97.1%, and 90% respectively. Overall agreements for these method were 93.5%, 97.4%, 86.8%, these results suggested that the urinary antigen detection test had high sensitivity and specificity. Our study indicated that concentrated urine samples increase sensitivity. We also evaluated the capabilities of both EIAs to detect soluble antigens were extracted from bacterial suspension of 18 strains of 5 Legionella species by heating. Both assays detected L. pneumophila serogroups 1 to 14, L. bozemanii. The Binax EIA proved to be useful as the Biotest EIA for diagnosis of legionellosis caused by Legionella species and serogroups other than L. pneumophila serogroup 1. Some cases have been shown to excrete antigen for prolonged period of times despite recovery from infection, so that the patient's history should be sought. The urine antigen detection EIA methods proved to be rapid and easy to use, detect antigen in the early stage of the disease with high sensitivity and specificity. Its use for the definition of legionellosis should be considered in Japan. PMID- 10386022 TI - [A trial of povidone-iodine (PVP-I) nasal inhalation and gargling to remove potentially pathogenic bacteria colonized in the pharynx]. AB - OBJECTIVE: Aspiration of potentially pathogenic bacteria (PPB) colonized in the upper airway is a major cause of bacterial pneumonia. We hypothesized that PVP-I nasal inhalation is effective in removing PPB from the upper airway. The aim of this study was to investigate the effectiveness and safety of PVP-I nasal inhalation. METHODS: Patients with asymptomatic PPB (MRSA and/or aerobic GNB i.e. Pseudomonas aeruginosa, Enterobacteriaceae) colonization in the pharynx were enrolled in this study. These patients were divided randomly into two groups as follows: a PVP-I nasal inhalation group (N group) which was asked to inhale 1% PVP-I solution x 2/day nasally by a jet nebulizer and gargling with PVP-I solution x 2/day, and a control group (C group), which was asked to gargle with PVP-I solution x 2/day. The study period was 2 weeks in both groups. RESULTS: Group N consisted of 16 cases, which included 9 (56%) cases with chronic respiratory complications and group C consisted of 14 cases which included 6 (43%) cases with complications. In N and C group, PPB disappearance from the pharynx was observed in 44% and 14% of patients after the study period, respectively. In the patients of group N, without chronic respiratory complication, PPB disappeared in 86% ot the cases. There was no adverse effect correlated with PVP-I nasal inhalation. CONCLUSION: We conclude that PVP-I nasal inhalation is a safe procedure for removing PPB from the upper airway, and this method may contribute to preventing bacterial pneumonia. PMID- 10386023 TI - [Biochemical and molecular characterization of Salmonella ser. enteritidis phage type 1 isolated from food poisoning outbreaks in Tokyo]. AB - Since the first outbreak in 1990, the incidence of Salmonella ser. Enteritidis (S. Enteritidis) phage type (PT) 1 food poisoning has gradually increased in Tokyo and has reached approximately 30% of the total S. Enteritidis outbreaks reported. To characterise these S. Enteritidis PT1 food poisoning, a total of 198 strains obtained from 44 outbreaks between 1990 and 1996 were examined for antimicrobial resistance, acid producibility from glycols (propylene and ethylene glycol) and plasmid DNA profiles. The 44 PT1 outbreaks analysed were further subdivided into 11 types by epidemiological markers. The most common patterns were type A (plasmid profile carrying only one plasmid (60 kb). SM and TC resistance and non producibility from glycols), and type B (plasmid profile carrying two plasmids (60 and 20 kb), SM resistance and no producibility from glycols) and were responsible for 21 (47.7%) and 15 (34.1%) outbreaks, respectively. In 11 of 44 outbreaks, strains carrying identical epidemiological markers were isolated both from patients and vehicle foods, environments, and/or food-handlers. Similar to PT4 and PT34 outbreaks reported in Japan, egg and egg related foods were also suspected in 8 of these 11 outbreaks. Of interest, chicken which were not pointed out in PT4 and PT 34 outbreaks was also suspected as a vehicle of transmission in two outbreaks. PMID- 10386025 TI - [Detection of bactericidal antibody in the breast milk of a mother infected with enterohemorrhagic Escherichia coli O157:H7]. AB - A 21 years-old pregnant woman developed diarrhea, fresh bloody stools and abdominal pain on April 6th 1997 at 32 weeks of gestation, and was admitted to the hospital on April 11th. The stool culture on admission was positive for enterohemorrhagic Escherichia coli (EHEC) O157:H7 (Stx1 and 2). Clinical laboratory data during admission showed only slight elevation of beta microglobulin and N-acetyl glucosaminidase in the urine, and no neurological or hemolytic symptoms were seen. After the antibiotic and lactobacillus administration, all her symptoms were relieved and no abnormal findings in pregnancy were observed. She delivered a baby girl normally on May 30th. Serum (between 41 and 120 days from the onset) and milk (between 4 and 64 days post partum) samples from the mother, and serum (64 days of age) from a baby and cord blood were obtained to monitor the immune status against EHEC O157:H7 and against Shiga toxins (Stx). Anti-E. coli O157 LPS antibodies (IgA, G and M) were assayed by the ELISA method. Neutralizing anti-Stx antibodies were measured by using ACHN cell cytotoxicity assay. In the colostrum and mature milk, high levels of IgA and IgM, and no IgG antibodies against EHEC O157 LPS were detected. In one of the control colostrum samples obtained from 4 healthy mothers IgA antibody against EHEC O157 LPS was detected. To assess the potency of protection against EHEC O157:H7 by the breast milk, we monitored it by the bactericidal activity for the organism under complement-coincubation experiment, and by the neutralization test for the Stx cytotoxicity. As a result, breast milk samples (both colostrum and mature milk) from a patient were demonstrated to kill the organisms. One of 4 healthy milk samples, showed bactericidal activity though it was negative in O157 LPS antibody. This bactericidal activity seen in one healthy colostrum is possibly due to a nonspecific reaction caused by non-O157 E. coli infection. From these observations, it was suggested that the bactericidal activity was due to the IgM class antibody against EHEC O157:H7. However, the neutralizing antibody against Stx1 and 2 could not be detected in any sample. EHEC infection at late gestation did not cause adverse effects to a fetus, and breastfeeding may have advantage for the protection of a baby against EHEC infection. PMID- 10386024 TI - [Prevalence and serotypes of verocytotoxin-producing Escherichia coli (VTEC) isolates from dairy cattle]. AB - To clarify the source of infection and route of transmission of Verocytotoxin producing Escherichia coli (VTEC) in humans, we collected fresh feces from healthy dairy cattle reared in Hokkaido, Fukushima, Kanagawa and Okinawa prefectures between June 1996 and March 1997, and attempted to isolate VTEC. The results are described below. 1) VTEC was isolated from 68 (27.1%) of 251 fecal samples tested. VTEC was isolated from 14 (28.0%) of 50 in Hokkaido, 13 (26.0%) of 50 in Fukushima, 20 (39.2%) of 51 in Kanagawa and 21 (21.0%) of 100 in Okinawa. There were no difference in the prevalence among the prefectures. 2) Toxin type and serotype of 85 isolates were determined. Thirty-three isolaties (38.8%) were classified into VT1 toxin and VT2 toxin, respectively, and 19 isolates (22.4%) were classified as the strain that produces both VT1 and VT2 toxins. The toxin types of these isolates were divided by serotypes. The VT1 producing isolates were the most frequent among O111:H-. The VT2-producing isolates included O2:H12, O2:H29, O2:H-, O82:H8, O82:HUT, O153:H19, O153:H42 and O153:H-. Among the isolates producing both VT1 and VT2 toxins, O153:H19 was relatively frequent. Based on findings that many bacterial strains coinciding with toxin types and serotypes of human-derived VTEC isolated from dairy cattle, it was suggested that dairy cattle are closely related to VTEC infection in human as a source of infection. PMID- 10386026 TI - [Assay of specific anti-Chlamydia pneumoniae antibodies by ELISA method. 3. Setting of serological criteria]. AB - "HITAZYME C. pneumoniae" (or "HITAZYME CPN", for short) is a diagnostic reagent that has been recently developed by adopting an ELISA method for detection of anti-Chlamydia pneumoniae (C. pneumoniae) antibodies. When this reagent is used under a current diagnostic standard that has been set as a provisional standard, however, high antibody positive rates are often produced for both IgG and IgA even using the specimens of healthy persons. So, it is difficult to distinguish C. pneumoniae-infected patients from healthy persons. Therefore, this time, we tried to establish a new diagnostic standard by setting up of special cut-off values for a single serum and rise rates of antibody titers for paired sera to improve the accuracy for diagnosis of C. pneumoniae infection. For a single serum testing, we set a special cut-off value at ID 3.00 for both IgG and IgA, so that most healthy persons fall within the range of the "negative" zone. This value was based on the calculation of "Mean+2SD" using measurement results (or IDs) of healthy persons. When this cut-off value was applied, the rate of > or = ID 3.00 for either IgG or IgA was 7.6% for healthy persons, and 64.9% for infected patients. (The rate reached 76.4% when the highest IDs of multiple specimens taken from each patient for this test were used in calculation) As a diagnostic standard for a single serum, therefore, it was defined that: "If ID is 3.00 or greater for IgG and/or IgA, it is highly likely that the case has an acute or a present infection." Using paired sera, we could confirm almost a linear relationship between the results by HITAZYME CPN and those by micro-IF method. Under micro-If method, if the antibody titer increases four times or greater using paired sera, acute infection is diagnosed. As it was found that the four fold increase in antibody titer corresponds to the increase of 1.35 in ID for IgG and 1.00 for IgA, we defined a diagnostic standard for paired sera as follows: "If ID increases by 1.35 or greater for IgG, and/or if ID increases by 1.00 or greater for IgA, the case may be diagnosed as acute infection." PMID- 10386027 TI - [Detection of Clostridium difficile toxin A from stool specimens by an enzyme immunoassay kit]. AB - Toxin detection from stool specimens is prerequisite for Clostridium difficile associated diarrhea and colitis. However, in Japan only one toxin detection kit is commercially available, which requires computerized VIDAS fluorescence reader. In this study we evaluated ImmunoCard Toxin A, which is an enzyme immunoassay with a format of individual cassette and needs no special equipment to perform, by comparing with the VIDAS CDA kit. Of 61 stool specimens 12 were positive and 39 were negative by both assays, 7 were VIDAS positive-ImmunoCard negative, 2 were VIDAS invalid-ImmunoCard negative, and 1 was invalid by both assays. Stool specimens which gave inconsistent results between two assays were subjected to cell culture assay to detect toxin B and culture for C. difficile followed by testing of toxin producibility of isolates. Of 7 VIDAS positive-ImmunoCard negative specimens 5 were negative for cell culture assay and toxigenic C. difficile culture and 2 were positive for cell culture assay. By omitting 3 VIDAS invalid specimens and counting 5 specimens, which were cell culture negative but VIDAS positive, as negative and 2, which were cell culture positive and VIDAS positive, as positive, 12 of the 14 VIDAS positive specimens were ImmunoCard positive (sensitivity, 85.7%) and all 44 VIDAS negative were ImmunoCard negative (specificity, 100%). These results indicate that although in comparison with VIDAS CDA, ImmunoCard Toxin A is slightly less sensitive, ImmunoCard is a rapid, simple, and reliable C. difficile toxin A detection kit, which has the advantage of requiring no special equipment to perform and no centrifuge step, as small as 25 muL of a specimen, and as short as approximately 15 min of time to complete the whole testing. PMID- 10386028 TI - [A case of lymphocyst infection caused by vanB type VRE]. AB - A case of post-operative abdominal lymphocyst infection caused by vanB type vancomycin resistant enterococcus (VRE) in reported. A 27-year-old female was diagnosed as pregnancy with uterine cervical carcinoma and underwent Cesarean section and radical hysterectomy. After discharge, she developed a high fever which was diagnosed as a lymphocyst infection. Microbiological examination demonstrated the presence of vanB type VRE in the cyst fluid. Cyst cleaning and minocyclin injection were effective. This is the first case of VRE infection in Japan. PMID- 10386030 TI - [Pulmonary infection caused by Mycobacterium gordonae]. AB - A 57-year-old woman who had been operated on for colon cancer and given chemotherapy, presented in September 1995 with worsening cough and abnormalities on her chest X-ray film. Acid-fast bacilli were isolated from the sputum. The organism was classified as M. gordonae by biochemical tests and DNA/DNA hybridization. The patient was treated with rifampicin and clarithromycin. Subsequently, sputum cultures became negative and the chest x-ray film showed a decrease infiltration. The findings in the present case suggest that M. gordonae may cause pulmonary infection and should be considered as an opportunistic pathogen. PMID- 10386029 TI - [A case of systemic lupus erythematosus complicated by Nocardia farcinica]. AB - We report a patient with systemic lupus erythematosus (SLE) complicated with nocardiosis. This case is very important that the complication of nocardiosis in SLE is very rare and the treatment to both SLE and nocardiosis is very difficult. A twenty-one-year old female was admitted to our hospital because of thoracic empyema and active lupus nephritis. Her medical history revealed that the diagnose of SLE was made when she was 18 with lymphocytopenia, proteinuria, positive antinuclear antibodies, and high titer of antibodies to native DNA. She was treated with prednisolne 60 mg daily and became better. Proteinuria appeared again in September 1995 and she was admitted to the former hospital. Renal biopsy proved diffuse proliferative glomeluronephritis (WHO IVb). She was treated with 1 g per day of methylprednisolone for 3 days and succeeded with 60 mg day of prednisolone. In early November she developed left chest pain and fever and chest X-ray demonstrated left pleural effusion. Antibiotics, antituberculosis, and antifungal therapy failed to subside her pleuritis and it turned to empyema. Then she was transferred to our hospital for further treatment. Nocardia farcinica was detected from the aspirated pleural fluid obtained at the former hospital. Drainage and intrathoracic impenem injection were effective. While long usage of minocycline was continued for the nocardiosis, 500 mg of cyclophosphamide pulse therapy to lupus nephritis was administrated. Two weeks later a new pulmonary lesion with left chest pain and liver abscess developed. Administration of trimethoprim-sulfamethoxazole subsided the nocardiosis. She was discharged with 1 g per day of proteinuria the prescribed 13 mg per day of prednisolone and continuous TMP-SMZ intake for nocardial infection. When immunosuppressive therapy must be given to the immunocompromised host, a more potent therapy must be added to avoid infection. PMID- 10386031 TI - [The prevalence survey on chronic tuberculosis patients with bacilli discharge in Osaka prefecture]. AB - This survey was made at the end of 1996 in Osaka prefecture including three ordinance designated cities of Osaka, Sakai and Higashiosaka. As of December 31 1996, 109 or 3.3% of active tuberculosis cases were found to be chronic tubercle bacilli excreters in Osaka city, and 128 or 3.6% in Osaka prefecture other than Osaka city, respectively. In the area called Airin at Nishinari-ward, Osaka, which has been an area with high prevalence of the disease, 33 or 5.8% were chronic tubercle bacilli excreters, and the rate was slightly higher than the other areas. Compared to the survey conducted 10 years ago, while the number of the chronic tubercle bacilli excreters decreased by half to 109 from 200 in Osaka city, to 128 from 211 in Osaka prefecture excluding Osaka city, the rate of chronic excreters to total active tuberculosis showed almost no change. Various factors such as difficult living conditions often attributed to defaulting of active tuberculosis patients from their treatment, thus resulting to development of chronic excretion. It is highly recommended to apply DOTS strategy for the completion of their treatment. Public health centers that have personal clinical records of each patient should be familiar with the results of their bacillus examinations. They should also be required to cooperate with medical institutions to cope with those who need retreatment. PMID- 10386032 TI - [Evaluations of MTD and Amplicor Mycobacterium for direct detection of Mycobacteria from clinical specimens]. AB - MTD (GEN-PROBE AMPLIFIED MYCOBACTERIUM TUBERCULOSIS DIRECT TEST) for Mycobacterium tuberculosis, and Amplicor Mycobacterium for Mycobacteria (AMP-M. tb for M. tuberculosis, AMP-M. av for M. avium and AMP-M. in for M. intracellulare) were used for the detection of relevant Mycobacterium. Their sensitivity and specificity were evaluated. Total 244 clinical specimens including 164 sputa were examined by the above two tests. The results were compared with those obtained by the conventional methods. Of 244 samples, number of the M. tuberculosis positive samples by microscopy, cultural test, MTD and AMP M. tb were 32, 33, 38 and 35, respectively. Among 33 culture positive samples, 25 were MTD positive and 26 were AMP-M. tb positive. Therefore, sensitivity of MTD and AMP-M. tb were 75.8% and 78.8%, and their specificity were 93.8% and 95.7%, respectively. When only sputa were used for the tests as the clinical specimens, both sensitivity of MTD and AMP-M. tb were increased to 94.4%. For MAC, positive samples of M. avium complex by culture, M. avium by AMP-M. av and M. intracellulare by AMP-M. in were 13, 16, and 8, respectively. Sensitivity and specificity of AMP-M. av/M. in were 100% and 95.2%, respectively. Clinical findings of the patients whose MTD tests were positive but negative by culture were reexamined. Three of 9 specimens were also positive in AMP-M. tb. From the records of the isolations of tubercle bacilli or other important pathogens from the other kind of clinical specimens, smear tests and patients' response to tuberculosis chemotherapy, four of 9 specimens were confirmed as true positive, three were suspected as positive, and two other specimens were false positive which might be caused by contamination. From these observations, it could be concluded that MTD and AMP-M. tb are more sensitive than conventional culture method, and MTD is more sensitive than AMP-M. tb but needs more careful treatment to avoid the contamination. PMID- 10386033 TI - [pncA gene mutations in clinical isolates of tubercle bacillus by polymerase chain reaction-direct sequencing method: in relationship to pyrazinamide resistance]. AB - We screened clinical isolates of tubercle bacillus for mutations in the pncA gene, which encodes pyrazinamidase (PZase), by polymerase chain reaction (PCR) direct sequencing method. Sixty-eight strains of tubercle bacillus were isolated from 32 patients with pulmonary tuberculosis. The patients were treated with antituberculous agents including pyrazinamide (PZA) for 2 months. Thirty-two of the 68 strains were isolated from sputum samples collected from the patients before treatment; 29 strains and 7 strains were collected after 1 month and 2 months of treatment, respectively. The pncA genes in these strains, were assessed for mutations by direct sequencing of PCR products using an automated sequencer. Similarly, we examined two clinical isolates (ka567 and minami22) of tubercle bacillus, determined to be deficient in PZase activity by the Wayne method. A PZA sensitive strain (H37Rv, ATCC27294), and a PZA-resistant strain (H37Rv-PZA-R, ATCC35828) were used as negative and positive controls for mutations in the pncA gene, respectively. None of the 68 strains demonstrated any mutations in the pncA gene; however, the 2 PZase-deficient strains had missense mutations in the pncA gene resulting in an amino acid substitution from His82 to Arg in clone ka567, and from Ala171 to Val in clone minami22. PMID- 10386034 TI - [A study of beta-lactamase activity of mycobacteria and clinical trial of penicillin/beta-lactamase inhibitor combinations in the treatment of drug resistant Mycobacterium tuberculosis]. AB - Beta-lactamase activity was determined using a nitrocefin disc method on 34 Mycobacterium tuberculosis (M. tuberculosis) strains and 13 nontuberculous mycobacteria strains. In the 34 M. tuberculosis strains, 23 strains showed beta lactamase activity. In 10 Mycobacterium avium complex strains, no beta-lactamase activity was detected. In the Mycobacterium chelonae strains, all three strains examined showed strong beta-lactamase activity. No correlation was found between beta-lactamase activity and resistance to anti-tuberculous chemotherapeutic agents. Four patients who were persistently positive for multi-drug-resistant M. tuberculosis (MDR-TB) on sputum and positive in beta-lactamase activity, were treated with penicillin/beta-lactamase inhibitor combinations. In two cases, the trials were discontinued because of diarrhea; the trials were continued in the remaining two for four months, but the MDR-TB was positive during the course of the therapy. Effectiveness of the therapy with penicillin/beta-lactamase inhibitor combinations against M. tuberculosis was obscure, although many of M. tuberculosis examined showed beta-lactamase activity. PMID- 10386035 TI - [A case of middle ear tuberculosis; PCR of the otorrhea was useful for the diagnosis]. AB - A 26-year-old female was admitted to our hospital with complaints of fever, cough, otorrhea and otalgia and progressive hearing loss of her left ear. Smears of her sputum were positive for acid-fast bacilli. Smears of her otorrhea were negative for acid-fast bacilli but PCR of her otorrhea was positive. Chest X-ray showed infiltrative shadows with the cavity. She was diagnosed as middle ear tuberculosis associated with pulmonary tuberculosis. After anti-tuberculous chemotherapy, fever, cough, otorrhea and pain of her left ear were improved, but her hearing level was not improved. In the case of middle ear tuberculosis, it is necessary to make an early diagnosis and treatment. This is the first reported case in Japan in which PCR of the otorrhea is positive. PMID- 10386036 TI - [The combination therapy of clarithromycin and sparfloxacin for pulmonary Mycobacterium gordonae infection]. AB - Seventy years old woman had fever and hemosputum at May 1997. She was diagnosed as mycobacteriosis because of the positive acid fast bacilli smear from sputum. Mycobacterium gordonae was isolated from sputum, gastric juice, and bronchial aspirate. The combination therapy of isoniazid, rifampicin, ethambutol, and clarithromycin was administrated; however, M. gordonae was not eradicated from sputum. Sparfloxacin was administered instead of isoniazid based on the result of drug susceptibility test. The smear became negative and M. gordonae was eradicated from sputum one month after the initiation of treatment with the combination of clarithromycin and sparfloxacin. PMID- 10386037 TI - [Serum angiotensin converting enzyme in patients with primary liver carcinoma]. AB - Recent studies have shown that serum activity of angiotensin-converting enzyme (ACE; EC 3.4.15.1) significantly decreases in patients with carcinoma of different localizations. There is no information in literature about measuring this enzyme in primary liver carcinoma patients. The serum activity of ACE has been examined on 15 primary liver carcinoma patients, 10 patients with cirrhosis, and 26 healthy subjects. Serum activity has been determined by spectrophotometric method using synthetic substrate Hip-His-Leu. The results were given in units which correspond to one nmol of hippuric acid released by enzymatic hydrolyze of Hip-His-Leu substrate in one minute on serum milliliter. The results have shown that serum activity of ACE increased in patients with cirrhosis (37.06 +/- 2.9; X +/- SEM; p < 0.05), and decreased in primary liver carcinoma patients (23.44 +/- 1.87; p < 0.01), what was statistically significant in comparison with the activity of the same enzyme in healthy subjects (29.90 +/- 2.72). These results point out the possibility of clinical application of measuring serum ACE activity as one of primary liver carcinoma marker in differential diagnosis of the disease. PMID- 10386038 TI - [Spontaneous abortion as an indication of degree of risk in pregnancy]. AB - The spontaneous abortions have a complex range of causes and consequences in connection with numerous biological and nonbiological factors. There are certain groups of women with the increased risk of unsuccessful reproduction. Some zygotes are genetically predetermined for abortion immediately or shortly after the conception. The cause, mechanism, pathology and frequency of such cases remain unknown, for the most part. PMID- 10386039 TI - [Endonuclease activity of recombinant pancreatic nuclease (A-K7H)]. AB - Pancreatic ribonuclease A (RNase A) is a endonuclease that catalyzes depolymerization of ribonucleic acid (RNA) releasing oligonucleotides. In the process of binding enzyme with substrate are involved several non-catalytic phosphate binding subsites, one of them is p2, additional to main catalytic site p1. RNaza A prefers binding and cleavage of longer substrate molecules, and 3',5' phosphodiester bond should be some six-seven residues apart from the end of molecules of the chain of RNA. In this work is analysed endonuclease activity of recombinant pancreatic RNase A (K7H), that in position seven instead of a lysine there is a histidine, amino acid residue that participates in main catalytic site p1. Mutant enzyme is obtained by site-directed mutagenesis by Kunkel. Results of this investigation have shown that substitution of lysine by histidine in position seven of RNase A has produced total deletion of p2 subsite, and K7H has lost endonuclease activity, and has become exonuclease. These results confirm central role of Lys-7 in establishing p2 subsite and endonuclease activity of pancreatic RNase A. PMID- 10386040 TI - [Optimization of HIV diagnosis using the RT-PCR (gag) method with various dilutions of cDNA and MgCl2 molarity]. AB - Lack of optimum conditions for PCR can lead to absence of desired PCR products, undefined multiplication and appearance of unwanted products. So, the use of PCR aiming to generate large amounts of target nucleic acid sequences, may be so called "double-edged sword". The important parameters in optimisation of PCR methodology are annealing temperatures, Mg++ concentration and different dilutions of target sequences. In our optimization experiments of HIV-RT-PCR (GAG) method we used HIV positive plasma specimens for extraction of RNA and production of cDNA by reverse transcriptase. Different cDNA dilution (10(-1)-10( 10)) and MgCl2 molarity (1.25 mM; 1.5 mM; 5.0 mM) we used for first round (GAG1 and GAG4 outer primers) and second round PCR (GAG2 and GAG3 inner primers). Optimal results after 3% NuSieve agarose gel electrophoresis and detection of 413 pb PCR products were obtained with 1.25 mM MgCl2 and cDNA dilution 10(-1) and 10( 2). So the main aim of PCR optimisation is the achievement of optimal primer template binding and primer extension. PMID- 10386041 TI - Late diastolic tumor "plop" in an asymptomatic case of right atrial myxoma. AB - Myxomas of the right atrium are rare tumors of the heart. They have been almost always described as symptomatic tumors. Auscultation and phonocardiography have been revealed tumor "plop" (P) as an early diastolic phenomenon, but the late "plop" has not be described. The aim of the case report is to present late diastolic tumor "plop" in an asymptomatic case of right atrial myxoma. A thirty year old man without any subjective symptoms was admitted to the clinic because of a murmur found during a routine examination when applying for a new job. Two dimensional echocardiography showed slightly enlarged right atrium with a myxoma in it. M-mode echocardiogram taken from parasternal short axis plane revealed a wide cluster of echoes in the right atrium moving into the right ventricle inflow tract in diastole. Simultaneous phonocardiogram showed splitting of the first sound (0.06 sec.). The tumor "plop" occurred in late diastole, 0.22 seconds after the second sound (S2) and coincided with maximum tumor protrusion into the right ventricle. After successful operation of the tumor without catheterisation, echophonocardiographic finding has become quite normal. PMID- 10386042 TI - [Trends in resistant bacteria isolated from a tube smear in intubated patients in intensive care]. AB - Long standing antibiotics therapy has resulted in growing bacteria resistance. We took a tube smear and prepared culture with antibiogram from the fifty intubated patients in the Intensive care unit in the war period. Gram-negative germs were the dominant ones in total sum, and among them the Acinetobacter calcoaceticus (No 21), Pseudomonas aeruginosa (No 18), Klebsiella pneumoniae (No 18), were isolated most frequently. Pseudomonas aeruginosa showed high resistance to Gentamicin (64%), Amikacin (35%), Trimethoprim (66%), Pefloxacin (20%), Ofloxacin (25%), Ciprofloxacin (25%). Klebsiella pneumoniae is resistant to Gentamicin (68%), Amikacin (22%), Cephalosporin (100%), Trimethoprim (31%), but it showed no resistance to chinolones. Acinetobacter calcoaceticus is resistant to Gentamicin (73%), Amikacin (36%), Cephalosporin (100%), Trimethoprim (63%), Pefloxacin (33%), Ofloxacin (67%), Ciprofloxacin (46%). Staphylococcus aureus was the most frequent among Gram-positive germs and it was resistant to Penicillin (100%), Gentamicin (40%), Lincocin (18%), Trimethoprim (5%), Pefloxacin (13%), Methicillin (21%), Cephalosporin (8%). The appearance of resistance on the antibiotics demands attentive follow-up aiming to influence the empirical application of antibiotics schemes depending on resistance. PMID- 10386043 TI - [Clinical aspects of multiple myeloma]. AB - This paper is a review of the patients with multiple myeloma, hospitalized at Clinic of Haematology in Sarajevo during the period from 1993 to 1998. This study encircles 45 patients; 18 males (40%) and 27 females (60%). Clinical and laboratory records, etiology, cytomorphology and radiography were analyzed in detail. The age of patients was 59.4 years (both males and females). The trends of disease showed increasing in 1994, 1997 and 1998, comparing it with other haematological malignancies (in 1993 the percent was 5.41, in 1998 was 15.83, and the average level, during the analyzed period, was 11.34%). The patients usually came in the terminal phase and that is the reason why median survival was 12 months. According to the results, the authors make a conclusion that there are some characteristics of this disease in individuals, comparing them with the results shown in relevant studies. They find the explanation in the exposure of the population to the chronic stress and deficit of energy caused by malnutrition during the aggression against Bosnia and Herzegovina. PMID- 10386044 TI - [Bronchobos in the therapy of chronic secretory otitis in children]. AB - In this study we showed our results in carbocisteine therapy of secretory otitis media in children. 66% of patients had successful treatment. We would recommend to make multicentric study with bigger number of patients to evaluate our results. PMID- 10386045 TI - [Subacromial pain syndromes as a possible results of errors in the initial diagnosis and therapy of the shoulder joint]. AB - The author in this paper shows possible diagnostic mistakes in the development of subacromial painful arc syndrome (cases with inadequate diagnostics). A hundred cases with acute shoulder's injury (without fractures and luxationes) who were admitted at the Clinic for bone surgery and Emergency Department of Clinical Centre, CUC Sarajevo in period between 1 January 1998-30 June 1998. Only clinical and X-ray examination in part were performed. A very interesting case with subacromial impingement as a consequence of such kind of treatment is presented in this paper. It can be concluded that it is necessary to use as a routine a comparative advantages of Echosonography, CT, MRI, as non invasive diagnostic methods at clinically suspected intra or extra articular lesions. PMID- 10386046 TI - [Results of in vitro fertilization therapy methods in Germany in 1996--the German In Vitro Fertilization Registry]. AB - We believe that on the IVF subject, which is nowadays applied exept a standard method in the world. In the next years we could expect further improvement in our success++. It is also to anticipate that next changes in the development of assistance-reproduction will be: 1. Freezing of ovums, 2. To get in vitro-meture of the immature cells, 3. Transplantation of ovarium tissue, 4. Transfer of the blastocyst, 5. Transfer of cytoplasma or the cell nucleus. We recommend that the IVF/ICSI--therapy and birth of children has to be registrated in one central register. PMID- 10386047 TI - [Protocol for management of normal pregnancy]. AB - On the basis of their experience and literature data the authors recommend "Protocol for antenatal care in normal pregnancy" with clear and detailed instructions for every examination during pregnancy. Protocol is based on 12 control examinations. At the first examination, when pregnancy is determined with certainty, blood pressure, body weight, pelvic diameters are to be measured and detailed data about personal and reproductive health history are to be taken and written in pregnancy card and patient's file. Specific attention is given to uterus size, cervix length and its dilatation. For the following check-up blood and urine samples, blood group, blood glucose level, Wassermann test, ultrasound examination are to be requested. Period between two check-ups is 4 weeks until 28th gestation weeks, than 3 weeks until 34th, 2 weeks until 38th and 1 weeks until labour. The last examination is planed for 40th week. Vaginal smear and urine sample testing should be controlled on every check-up. Blood picture and blood glucose level are to be checked three times in pregnancy. Ultrasound examination is planned for the first trimester, 20th and 34th gestation week. PMID- 10386048 TI - [The prevalence of smoking, alcohol consumption and drug abuse in school children and adolescents]. AB - In this work the authors have examined through the survey, the prevalence of smoking, alcohol consumption and drug abuse among the school children and youth. Subjects in this research were 208 pupils in Primary school and 232 pupils in Secondary school. It has been found that in the group of Primary school children 0.9% smoke, 5.83% consume alcohol and 0.9% use narcotics. Out of total number of subjects from Secondary school 27.97% smoke, 28.81% consume alcohol and 10.59% use narcotics. The problem is especially prominent among the boys and girls of the last two years of Secondary school: 42.86% smoke; 41.35% consume alcohol and 13.53 use narcotics. PMID- 10386049 TI - [Health education as a factor in decreasing traffic accidents]. AB - Traffic security in post-war Bosnia and Herzegovina is getting worse. Use of motor vehicles and frequency of roads are in constant increase. Import of cars of different types, models, ages and quality caused that we are among the countries with insufficiently developed traffic security. Critical group of the traffic participants consists of young people and children. Their behaviour in traffic depends on their knowledge and attitude gained at home and school. Goal of this paper is to point out the importance of health-education activities in traffic security development. Evaluation was performed among the school children in a transit area in Bosnia and Herzegovina. Results show that school children have no enough knowledge about traffic rules; 13.4% of them don't know a single traffic sign. Even 19.9% of them got injured in traffic accidents, out of which 41.4% got injured while riding a bicycle and 22.4% as pedestrians. These initial results show that level of children's traffic culture can be raised only through systematic and permanent education within regular curriculum. Topics on traffic security should be an integral part of education programme, and presented through various subjects. PMID- 10386050 TI - [Information technology as a support in the area of education]. AB - Decision Support Systems (DSS) are more and more used and developed, mainly within the management area. It is used at strategic level for global planning and managing the organization; at tactical level for short-term planning, process realization and control. Although many authors state that DSS are used only at global and tactical level, some of them--through practical applications--show that specific DSS can be created in order to support decision in other areas than classical management. Paper describes computer application that efficiently supports organisation of exams at The Department for Medical Informatics of Medical Faculty in Tuzla. PMID- 10386052 TI - [Molecular genetic mechanism of hereditary human kidney cancer development]. AB - Here we reviewed the molecular genetic mechanism in the development of 4 types of human hereditary kidney cancers. These include von Hippel-Lindau (VHL) disease, hereditary papillary renal carcinoma, familial renal cancers with translocation of chromosome 3, and Tuberous sclerosis. Loss of function of the VHL disease gene is responsible for the von Hippel-Lindau disease and major portion of sporadic clear cell renal carcinoma. Activated c-Met oncogene is responsible for the development in some cases of hereditary papillary renal cell carcinomas and sporadic papillary renal carcinomas. There are several cases of familial renal carcinoma in that translocations of chromosome 3 p are demonstrated. The molecular genetic mechanism of this disease is not known. Several reports show the development of renal cell carcinoma in Tuberous sclerosis patients. TSC 1 or TSC 2 gene may be responsible for these tumors. The detail in this disease not well known. Molecular genetic analyses for hereditary renal cancer identified several oncogenes and tumor suppressor genes in hereditary as well as sporadic renal carcinomas. Future studies may reveal new category of oncogenes or tumor suppressor genes that are involved in the human kidney cancer development. PMID- 10386051 TI - [Al-Biruni--a universal scientist]. AB - Al-Biruni's was of Persian descent. He was born in Horesmiya and had studied mathematics, history and medicine. Acquiring knowledge from these sciences, he wrote an outstanding work on chronology of several nations and devoted it to Ziyarit ruler Kabus. He made a chronological overview of calendars from many nations, including Persians, Greeks, Egyptians, Jews, Melkitian and Nestorian Christians, Sabeyaans as well as the old Arabs. Data presented in the work, according to the later authors, were taken from very reliable sources. He was contemporary of Ibn-Sina, and thanks to their friendship, they have discussed very much miscellaneous topics. He belonged to the group of scholars, taken by Gaznevian Soultan Mahmud to a long journey to India. Afterwards Al-Biruni wrote and published detailed work "Description of India"--a work on cultural history of India. Due to excellent abilities of Al-Biruni as a philosopher and scholar, there are still significant and reliable notes about buddhistic philosophy, structure of castes and Brahmans' life style. In this Al-Biruni's masterpiece, there are many comparative analysis of Suffism and certain Indian philosophical methods. Al-Biruni's most important work is "Pharmacopoeia"--"Kitab al-saydala", which brilliantly describes all medicaments. This work has been published in many languages. He also wrote few works on astronomy and astrology. In those works he has explained some astrological events through scientific approach in a such peculiar way that nobody has ever explained before. He was also interested in sciences like geology, mineralology, geography, mathematics, psychology and many others. PMID- 10386053 TI - [Clinical study of radical retropubic prostatectomy for prostate cancer]. AB - PURPOSE: To evaluate the results of radical retropubic prostatectomy in patients treated at a single institution. MATERIALS AND METHODS: Between April 1985 and July 1997, 76 patients with prostate cancer underwent radical retropubic prostatectomy, including 73 receiving pelvic lymphadenectomy. The median age and follow-up time were 68 years old and 44 months, respectively. The pathological stage was pT0 in 6 patients, pT2 in 29, pT3 in 39, pT4 in 2, and pN+ in 22. RESULTS: The surgical margin was positive in 10% of the pT2 patients and 61% of the pT3 patients. Twelve patients had recurrence. Recurrence was shown by biological failure in 4 patients and clinical failure in 8. The disease-free 5 year survival rates (Kaplan-Meier) were 100% in pT0 patients, 87% in pT2, 72% in pT3, 50% in pT4, 77% in pN-, 75% in pN+, 73% for a positive surgical-margin, and 83% for a negative surgical-margin. There were no statistical differences between any of these factors. However, the disease-free survival rate in pT3 patients with poorly differentiated adenocarcinoma (PDA) who received postoperative radiotherapy combined with hormonal therapy was significantly superior to that in patients with the same characteristics who received hormonal therapy (100% vs 27%; p = 0.011). The cause-specific 5-year survival rates were 100% in pT0, 100% in pT2, 92% in pT3, 50% in pT4, 94% in pN-, 93% in pN+, 93% for a positive surgical-margin, 98% for a negative surgical-margin, 100% in the aforementioned pT3 patients with PDA and postoperative radiotherapy combined with hormonal therapy and 86% in pT3 patients with PDA and postoperative hormonal therapy. There were no statistical differences between any of these factors. CONCLUSIONS: Our results suggest that radical prostatectomy is available for both organ confined and non organ-confined advanced prostate cancer. Postoperative radiotherapy combined with hormonal therapy is especially useful for patients in pT3 with PDA. PMID- 10386055 TI - [The natural history of renal angiomyolipoma]. AB - PURPOSE: To make the policy of treatment with angiomyolipomas (AML) more clear, we discussed the natural history of angiomyolipomas retrospectively. PATIENTS AND METHODS: Between May 1982 and December 1997, 14 patients with AML in 18 kidneys were followed, who were 2 men in 2 kidneys and 12 women in 16 kidneys, 27 to 80 years old. No patients suffered from tuberous sclerosis. Symptoms, initial sizes and changes of the size were evaluated for these patients. RESULTS: Ten patients with AML in 14 kidneys were asymptomatic and four patients were symptomatic. But one of the 4 patients had symptoms of abdominal pain and palpable mass which were due to contralateral AML that were treated with nephrectomy, so symptoms due to small AML were seen in 3 cases (2.0 cm, 3.5 cm, 3.8 cm). Among 11 patients in 15 kidneys followed radiologically for more than 6 months, the tumors were unchanged in size in 7 kidneys, which were in all of 6 cases with unilateral solitary tumor and in 1 with bilateral multiple tumors. In other 8 kidneys the sizes of the tumors were increased, which were in the cases with multiple tumors in one kidney or in bilateral cases. Compared to the cases of unilateral solitary AML, the size of AML with multiple tumors in one kidney or in bilateral kidneys significantly increased (p < 0.01). Embolization were performed for 4 kidneys, which were in 2 cases with increased tumor in size to more than 4 cm in following period, in 1 with dull flank pain, and in 1 with the tumor more than 4 cm at diagnosis that grew to more than 5 cm. CONCLUSIONS: Unilateral solitary AML was appeared to be hard to increase in size and to have a different natural history from bilateral or multiple tumors. PMID- 10386054 TI - [Detection of prostate-specific antigen mRNA in preoperative peripheral blood of patients with prostate cancer: relationship to pathological parameters of the surgical specimens]. AB - PURPOSE: Up to 40% of surgically treated prostate cancers are bound to be understaged by using current diagnostic modalities. To improve on current staging methods for prostate cancer, we evaluated whether nested reverse transcriptase polymerase chain reaction (RT-PCR) for prostate-specific antigen (PSA) mRNA can improve preoperative staging of prostate cancer. MATERIALS AND METHODS: From May 1996 through November 1997, 30 patients with prostate cancer (T1-T3) were evaluated for PSAmRNA in their peripheral blood by nested RT-PCR before radical retropubic prostatectomy. RESULTS: A highly sensitive RT-PCR assay employed detected one PSA-expressing cell (LNCaP) diluted into ten million mononuclear cells. All 15 controls, including 9 women, showed negative PSAmRNA in their peripheral blood, whereas 16 of 30 (53%) patients with prostate cancer showed positive PSAmRNA in their preoperative peripheral blood. Interestingly, 7 patients with positive PSAmRNA had pathologically organ-confined prostate cancer. No significant relationship was demonstrated between positive PSA-PCR results and clinico-pathological parameters such as clinical stage, pretherapeutic serum PSA, pathological stage, seminal vesicle invasion, lymph node metastasis, vascular invasion, or Gleason sum. CONCLUSION: Our present study showed that the results of PCR-based PSAmRNA assay had no significant association with clinico pathological parameters. However, the prognostic significance of detecting the circulating prostate-specific signals requires a longer follow-up, which is currently under study. PMID- 10386056 TI - [Effects of ischemia on voiding function and nerve growth factor of the rat urinary bladder]. AB - PURPOSE: Nerve growth factor (NGF) is synthesized in the target organs innervated by autonomic and sensory nerves so as to grow, maintain and/or repair the neurons. The present study evaluated the effects of ischemia on NGF synthesis and voiding function of the rat urinary bladder. MATERIALS AND METHODS: Bladder ischemia was induced by ligating of bilateral internal iliac arteries in the female rats. We examined the changes in the blood flow, histological structure, voiding function and NGF content of the bladder immediately, 1, 7, 14 and 28 days after the surgery. Blood flow was estimated by measuring absorbance of homogenized bladder tissue after dye injection into the abdominal aorta. Voiding function was assessed by continuous cystometry under an awake restrained condition. NGF was quantified by enzyme immunoassay (ELISA method). RESULTS: Blood flow decreased to 18% of the control immediately after the vascular ligation, and gradually recovered to 66% of the control on day 28. Histologically, epithelial ablation and thinning of muscle layer were observed on days 1 and 7. These histological disorders gradually improved to normal appearance on day 14. On day 1, while the maximum contraction pressure significantly decreased, the contraction frequency and small prevoiding contraction increased. On the other hand, the voiding efficacy markedly decreased on day 7. These functional changes recovered nearly to the control levels after day 14. NGF content transiently increased 2.4 times as the control on day 1. CONCLUSION: The present results will indicate that the voiding function deteriorated by acute ischemia is temporarily compensated by detrusor hyperrefrexia, which may be attributable to an enhanced NGF synthesis, and then improves by the development of collateral blood circulation. PMID- 10386057 TI - [Long-term follow-up of a bladder exstrophy case after successful repair]. AB - We present a female case of bladder exstrophy where the patient was followed up for 15 years. In 1982 primary closure of the bladder and urethra with bilateral iliotomy by the posterior approach was performed at the age of 4 months. However, realignment was necessary, since the wound split open postoperatively. We used a corset specially prepared for this patient to prevent wound dehiscence during the subsequent postoperative course. Four years later, VUR was surgically eradicated. At the age of 9 years old, her bladder function was satisfactory, demonstrating a normal CM pattern with synergia and Pmax 128 cm H2O on UPP. She had no urge or stress incontinence. The most recent IVP and DMSA renal scan revealed almost normal findings. VCUG showed no VUR and renal function tests (PSP, Ccr 24) also confirmed no interval deterioration. We believe that the approximation of the intersymphyseal band at the time of bladder neck closure is the single most important factor for ensuring urinary continence. PMID- 10386058 TI - [Screening of prostate cancer with PSA and transperineal six sextant biopsy]. AB - OBJECT: The objectives of this study are to examine how many cancer patients we can detect among the outpatients whose PSA values are above 4.0 ng/ml, and to compare the usefulness of transperineal six sextant biopsy (ss-biopsy) with that of transrectal one. METHODS: All the male outpatients (above 50 years old) were inspected Tandem-R PSA levels and digital rectal examination (DRE). Among them, 129 patients showed more than 4.0 ng/ml of PSA values and/or positive finding of DRE, and underwent subsequent transperineal ss-biopsy. RESULTS: Cancers were detected in 52 patients (40.3%) without major complications. Among 64 gray zone (PSA 4.1-10.0 ng/ml) patients, 17 (26.6%) were found to be cancer by ss-biopsy, meanwhile only 2 cancer patients (8.9%) were detected from 23 gray zone ones by traditional directed biopsy. Application of PSA density could not be found practicable to eliminate unnecessary biopsies in the gray zone group. CONCLUSION: Prostate cancer could be found nearly a fourth in the gray zone group of the outpatients. To enhance the detection rate, obtaining at least 6 core samples are recommended from either perineal or rectal root. PMID- 10386059 TI - [Dermoid cyst with thyroid follicle perforated into bladder and ileum: a case report]. AB - A 64-year-old woman was admitted to our hospital on August 19, 1996 with the chief complaint of microscopichematuria. A solid mass was found in her lower abdomen. An abdominal CT scan suggested a large intrapelvic cystic mass and the existence of a fistulous connection between the mass and the small intestine. The existence of a fistula was confirmed by a preoperative barium enema and a cystscopic study. On September 25, 1996, a suprapubic partial cystectomy and total hysterectomy were performed since an intrapelvic abscess was suspected. The cystic mass was observed to adhere to the hollow viscus (uterus, rectum, appendix, ileum and bladder). As a result a part of the ileum and bladder were also removed with the mass. The contents of the cyst included a foul smelling gas and white-green pus. Using a probe, we found two fistulous openings to the ileum and bladder. The histopathological findings indicated a dermoid cyst of the left ovary with thyroid follicles, which are known as "Struma ovarii". PMID- 10386060 TI - [Fatal hepatic failure following hepatitis caused by flutamide: a case report]. AB - A seventy-three year-old patient with prostate cancer underwent radical prostatectomy, followed by total androgen brocking therapy using flutamide and LH RH agonist. As Hepatic dysfunction (GPT = 3.045 IU/l) was noticed by periodic blood analysis, flutamide was stopped and he was hospitalized immediately without any subjective symptoms. Ten days after the admission, he developed massive bleeding from duodenal ulcer, resulting in duodenal perforation. Following the emergency operation, plasma exchange therapy was repeated against serious hepatic dysfunction. However, he was dead of pneumonia two months after the admission. Autopsy revealed biliary congestion in a small liver, although it was not cirrhotic. In our patient, hepatic dysfunction was irreversible and prolonged. We strongly recommend to perform serial liver function test from the start of treatment with flutamide, especially during the initial three months. Flutamide should be stopped promptly it significant liver abnormalities are detected. PMID- 10386061 TI - [In focus: Standard combination in asthma therapy. RPR-Press Conversation at the 40th Congress of the German Society for Pneumonology, Bad Reichenhall, 19 March 1999]. PMID- 10386062 TI - Basic principles of gene transfer in hematopoietic stem cells. PMID- 10386063 TI - Optimizing conditions for gene transfer into human hematopoietic cells. PMID- 10386064 TI - Transfer of the MDR-1 gene into hematopoietic cells. PMID- 10386065 TI - O6-benzylguanine-resistant mutant MGMT genes improve hematopoietic cell tolerance to alkylating agents. PMID- 10386066 TI - Use of variants of dihydrofolate reductase in gene transfer to produce resistance to methotrexate and trimetrexate. PMID- 10386067 TI - Augmentation of methotrexate resistance with coexpression of metabolically related genes. PMID- 10386068 TI - Protection of bone marrow cells from toxicity of chemotherapeutic agents targeted toward thymidylate synthase by transfer of mutant forms of human thymidylate synthase cDNA. PMID- 10386069 TI - Dihydropyrimidine dehydrogenase and resistance to 5-fluorouracil. PMID- 10386070 TI - Chemoprotection against cytosine nucleoside analogs using the human cytidine deaminase gene. PMID- 10386072 TI - In vivo selection of genetically modified bone marrow cells. PMID- 10386071 TI - In vivo selection of hematopoietic stem cells transduced with DHFR-expressing retroviral vectors. PMID- 10386073 TI - Gene transfer in the nonmyeloablated host. PMID- 10386074 TI - Methods in outcomes research in hepatology: definitions and domains of quality of life. AB - The health care system in the United States is at a turning point. We are experiencing a shift in focus from structure and process to outcomes, and from specific clinical outcomes to generic outcomes. It is no longer sufficient to document where and how care is delivered and what clinical indicators were changed through various interventions. Rather, payers, employers, and patients themselves demand evidence that the health care system produces patients who feel better, can do more, and are confident about their health. These concepts, collectively referred to as Health-Related Quality of Life (HQL), are an important aspect of the natural history of liver diseases and an important means of assessing the results of therapeutic interventions. PMID- 10386075 TI - Validation of health-related quality of life instruments. AB - The development of instruments to quantitate health-related quality of life (HRQOL) includes taking the appropriate and, in fact, essential steps to ensure that the designated tool measures what it purports to measure and does it accurately and reliably. Such steps are not only critical in the development of appropriate tools, but, when clearly presented, also serve to prove to the reader (and reviewers) that the instrument is a legitimate one. Key to the development of any HRQOL instrument are three fundamental concepts: validity, reliability, and responsiveness. Validity proves that the instrument measures what it claims to measure. Reliability proves that the results are reproducible in a stable setting. Responsiveness proves that the instrument is sensitive enough to record important differences or changes. Each of these principles is discussed, including some examples of the application of these principles in previously published HRQOL studies. The studies referenced are excellent examples of the methods used in the creation of a HRQOL scale, and the reader is strongly encouraged to review these in their entirety to fully grasp the concepts. PMID- 10386076 TI - Strategies and pitfalls in quality of life research. AB - QOL research is an important element of outcomes research and can provide an important adjunct to clinical trials in hepatology. The principles of QOL research are similar to all outcomes research methods. Pitfalls in QOL research include poor study design, choosing the wrong instrument, and inadequate interpretation of design. These pitfalls can be avoided with a well-thought-out strategy including a well-designed hypothesis to test, a validated instrument, careful data collection and analysis, and collaboration with biostatisticians and experts in health-utility research when necessary. QOL research is best suited to questions where survival differences between the two groups studied are modest or to balance economic outcomes, but it can be a useful part of virtually any clinical question. PMID- 10386077 TI - Economic considerations for the hepatologist. AB - This is an era of rapid change in health care systems and clinical practice. In the face of increasing national health care expenditures, physicians are confronted with an increased demand to justify practices and to show the value of their services. Hepatologists are being required to show that their practices are both effective and cost-effective. This has led to an expanding body of literature examining the cost-effectiveness of medical practices. To evaluate these economic analyses the reader must be familiar with the concepts used in economic analysis and have a clear understanding of both how these analyses are performed and how the results can be applied to clinical practice. The purpose of this article is to provide the reader with the essential concepts for evaluating economic analyses in the medical literature and to provide published criteria for performing and critiquing an economic analysis. The terms used in economic analysis are outlined and defined. The criteria for performing an economic analysis are listed. Examples are given to emphasize the key points. PMID- 10386078 TI - Analytic approaches for the evaluation of costs. AB - Currently, economic evaluation of new medical therapies is conducted routinely. Development of an analysis plan before performance of the analysis is a first step in the analysis of data from such evaluations. Univariate analysis of costs can be performed with both parametric and nonparametric tests. Potential multivariable analyses include ordinary least squares regression, nonparametric hazard models, parametric failure time models, Cox semiparametric regression, and joint distributions of survival and cost. In addition to developing point estimates for economic outcomes, 95% confidence intervals for cost-effectiveness ratios should be developed to evaluate the level of uncertainty that surrounds these estimates. Sensitivity analysis should be used to address other sources of uncertainty. PMID- 10386079 TI - Challenges in economic outcomes research. AB - In light of current pressures to limit health care expenditures, economic evaluations will continue to be conducted to help assess the value of new therapies or treatment programs. Although guidelines for the conduct of economic evaluations have been proposed, it is unlikely that a standard/common methodology will (or should) be used. Consequently, it is important that the reader/reviewer of such analyses be aware of the potential challenges or difficulties when evaluating these assessments. While not providing a comprehensive list of these challenges, it is hoped that this article will serve to bring awareness to some of these challenges in the design, analysis and interpretation of these evaluations. PMID- 10386080 TI - Meta-analysis: when and how. AB - Systematic reviews have a central role in evidence-based medicine. The quantitative systematic review, also known as meta-analysis, provides a logical structure for quantifying evidence and for exploring bias and diversity in research systematically. It is essential that clinicians, educators, and researchers understand the methods that comprise this research tool, particularly the basic step-by-step process, and know when numerical pooling of data is appropriate. The essay describes how systematic reviews are best conducted and when statistical pooling of data is appropriate. Systematic reviews are scientific investigations with planned methods that use original studies as subjects and synthesize the results of multiple studies using strategies to limit bias and random error. This process requires judgments to be made explicit, and should be question driven, protocol based, reproducible, and comprehensive in scope. Meta-analysis provides a framework for research synthesis, increases power and precision, provides an overall estimate and range of effect, and identifies greater-than-expected variability among study results (heterogeneity). Meta analysis does not remove subjectivity from the process of synthesis, identify sources of variability among studies, or obviate the need for sound, compassionate clinical reasoning. Statistical heterogeneity should be anticipated and welcomed. It forces a consideration of clinical heterogeneity as well as variation in study protocol and quality. Statistical tests for homogeneity are insensitive and do not indicate sources of heterogeneity, making such consideration imperative. The most common and popular measures of efficacy for a meta-analysis are the standardized difference between two means, the relative risk, and the odds ratio. An additional measure, the number needed to treat, with its 95% confidence interval is the most clinically useful measure of the effects of an intervention and is useful for comparing the relative effectiveness of different interventions for the same condition. Important parts of meta-analysis and sensitivity and subgroup analyses are best considered a priori and should be used to explore heterogeneity and to test for publication bias and variation in study quality. PMID- 10386081 TI - Neural networks in outcomes research. AB - Neural networks (NNs) are being used in the areas of prediction and classification of outcomes in medicine--areas in which regression models have traditionally been used. In this report, we summarize the steps in developing and testing an NN. Through applications in clinical research, we present several examples of NN development and evaluation. Through these applications we show that the performance of the NNs matched or exceeded the performance of traditional methods. We then discuss the advantages and disadvantages of NN models in comparison to traditional regression methods. PMID- 10386082 TI - Using outcomes data to identify best medical practice: the role of policy models. AB - Increasingly, physicians are attempting to incorporate best evidence into their clinical decision making. However, best evidence takes a variety of forms, including clinical trials, cohort studies, administrative data, and patient preference data. Incorporating multiple data sources in a way that informs complex clinical decisions is a substantial analytical challenge. One approach to this challenge is to develop a simulation/decision model that explicitly represents the natural history of disease and the impact of treatments on that natural history. The model should be requisite--that is, sufficient in form to address the decision problem--but not overly complex. Such a model can be of value because it (1) allows a variety of viewpoints to be considered, (2) incorporates the best scientific evidence, and (3) permits sensitivity analyses to evaluate the impact of alternative clinical scenarios and uncertainty in model inputs. The Stroke Prevention Policy Model (SPPM) illustrates this approach. The SPPM is a simulation model designed to predict the best among various treatment alternatives for preventing strokes. Similar models can be applied to treatment outcomes for liver disease. PMID- 10386083 TI - Ensuring the success of URM medical students. PMID- 10386084 TI - Ensuring routine attention to advance directives. PMID- 10386085 TI - Leadership experiences of conditional early-acceptance students. PMID- 10386086 TI - Learning disabilities, medical students, and common sense. PMID- 10386087 TI - Physicians and attorneys: a non-meeting of the minds. PMID- 10386088 TI - Mission-based budgeting: removing a graveyard. AB - Many activities in today's medical schools no longer have medical students' education as their central reason for existence. Faculty are hired primarily to provide clinical service or to make discoveries, with the role of educator of secondary importance. Budgeting in medical schools has not evolved in concert with these changing roles of faculty. The cost of medical students' education is still calculated as if all faculty were hired primarily to teach medical students and their other activities were to support this "central" mission. Most medical schools still mix revenues without regard to intent and cannot accurately determine costs because they confuse expenses with costs. At the University of Florida College of Medicine, a group of administrators, chairpersons, and faculty developed a budgeting process now called mission-based budgeting. This is a three step process: (1) revenues are prospectively identified for each mission and then aligned with intended purposes; (2) faculty productivity, i.e., faculty effort and its quality, is measured for each of the missions; and (3) productivity is linked to the prospective budget for each mission. This process allows the institution to understand the intent of its revenues, to measure how productive its faculty are, to learn the true costs of its missions, to make wise investment decisions (subsidies), and to justify to various constituents its use of revenues. The authors describe this process, focusing particularly on methods used to develop a comprehensive database for assessment of faculty productivity in education. PMID- 10386089 TI - One hospital's successful 20-year experience with physician assistants in graduate medical education. AB - The downsizing of residencies and the migration of residents to outpatient settings create an increasing need to protect residents' educational experiences and to maintain standards of hospital care. Some hospitals have solved this dilemma by using mid-level practitioners (MLPs), including physician assistants (PAs), to augment the diminished staffs of residents in their surgical residencies. The authors describe how their hospital has done so. Their surgical PA program, begun in 1979, seeks to meet the hospital's expectations for in-house coverage of surgical patients, to protect the educational integrity of the physician residency program in surgery, to allow protected time for residents' conferences and clinics, and to prepare residents for future practice in multidisciplinary teams. The PA and residents' services are partly separated, which reduces the potential for resident-PA conflict. Responsibilities for both residents and PAs are stratified (junior vs senior status). Both services are teaching services, which helps motivate PAs to be committed to the service and helps foster the equality between residents and PAs that the program strives for. The residents have come to value the PAs, and the program's goals have been achieved, including protecting time for residents' education and maintaining humane on-call schedules for residents. The authors discuss job satisfaction, turnover, and the hard financial realities of paying for PAs' salaries, benefits, and educational programs, as well as the loss of Medicare DME and IME reimbursements when a PA replaces a resident. Ways some of these costs can be recovered are outlined. The authors conclude with recommendations on how to deal with six key issues of PA or other MLP programs: need for institutional commitment; importance of local circumstances; emphasis on partnership, not competition, between PAs and residents; value of an education component; need to build a cohesive program, and the importance of effective PA leadership. PMID- 10386090 TI - Scholarship, humanism, and the young physician. AB - It is time to understand the value of a broad liberal education for those college students who aim to be physicians, both because the medical curriculum is becoming more humanistic (which a liberal education would support) and because three enormous challenges confront physicians and educators alike: the relentless tide of biomedical discoveries, the great financial burden that medical care imposes, and the public's desperate plea for physicians who are more caring and communicative. A liberal education--meaning a course of study that is largely unrestricted and that attempts to sample the entire breadth of human knowledge- can help the premedical student cultivate, ripen, and enrich fundamental proficiencies such as accurate recording of observations, communicating ideas well, dealing with human emotions and becoming sensitive to human frailties, learning to listen and respond appropriately, learning to make sound judgments, and cultivating empathy and compassion. These are all skills that a liberal education can help the young student learn early rather than late, skills that prepare the student for dealing later with complex social, ethical, and clinical issues as a physician. A liberal education also can help prepare the student to take advantage of other general educational opportunities that are available in the small, closed community of a residency, such as learning to both assume and delegate responsibility, to participate in rational debate while respecting the opinions of others, and to exercise mature judgment, civility, empathy, and compassion. While a liberal education will not necessarily make the student a more technically proficient doctor, for some it will be essential to awaken and sharpen those essential skills that a physician needs to rise to the top of a profession that never fails to recognize excellence and humanity. PMID- 10386091 TI - Rating educational quality: factors in the erosion of professional standards. AB - Changes in the health care environment are putting increasing pressure on medical schools to make faculty accountable and to document the quality of the medical education they provide. Faculty's ratings of students' performances and students' ratings of faculty's teaching are important elements in these efforts to document educational quality. This article discusses selected research related to factors affecting raters' judgments, analyzes how changes in the health care environment are influencing such judgments, and links these influences to the system that upholds professional standards. Ratings are known to have a positive bias (generosity error), provide limited discrimination, and often fail to document serious deficits. The potential sources of these problems relate to the mechanics of the rating task, the system used to obtain ratings, and factors affecting rater judgment. As managed care demands reduce the time faculty have for teaching, as system-wide disincentives to provide negative ratings proliferate, and as social engineering challenges, such as the Americans with Disabilities Act, impose differential standards for students, the natural tendency to avoid giving negative ratings becomes even harder to resist. Ultimately, these forces compromise the capability of faculty to uphold the standards of the profession. The author calls for a national effort to stem the erosion of those standards. PMID- 10386093 TI - Bench to bedside in medical education. AB - In spite of the significant resources devoted to medical education at all levels, the study of educational process has received relatively little attention. Most studies of learning have been observational, with little reference to recent advances in the neurobiology of learning and memory. In this article, the author briefly reviews developments in our understanding of the neurobiology of learning and memory and suggests ways they could contribute to the future evolution of medical education. PMID- 10386092 TI - Interdisciplinary ambulatory education and service in primary care at the University of New Mexico. AB - To compete and survive in a managed care market, academic health centers must develop integrated delivery systems in general and an integrated primary care system in particular. However, the departmental-based structure at most academic health centers is ill-suited to this purpose. Service and education are usually segregated by department, and the professional activities of primary care faculty in different departments are fragmented, leaving them weakened as a political force within the institution. The University of New Mexico established a model of integrated primary care education and service by creating three interdisciplinary primary care clinics staffed by primary care residents and faculty. The clinics attracted a substantial portion of each department's faculty and residents. The clinics united primary care providers from different departments as a stronger, more unified voice in negotiating with the hospital and in fostering needed changes for primary care in the institution. Interdisciplinary teams require considerable time and labor both in planning (because of joint decision making) and in operation. Better staff structures and staff development must be learned through trial and error because there are not established benchmarks for interdisciplinary teams. Governance presents problems, primarily because loyalties to departments may supersede those to the clinic practice, and sometimes the departments' teaching priorities are challenged by clinic directors' need to ensure filling their interdisciplinary staff needs. These obstacles to collaboration can be addressed creatively, and ultimately the comprehensiveness and quality of care convinces providers and the institution. PMID- 10386094 TI - The Visible Human Project: a resource for education. AB - Throughout recorded history, the relationships between biological structure and function have been central to the understanding of health and disease. For many centuries, the anatomy illustrations originally created during the medieval period formed the basis for the study of medicine. But learning and understanding anatomic structures is limited by the fundamentally two-dimensional images traditionally used to teach them. The digital computer now allows scientists to acquire, store, manipulate, and display complex images. In 1989, the National Library of Medicine (NLM) began a project to use computer technologies to build a prototype digital image library of data representing a complete normal adult human male and female. In this paper, the author describes the beginnings of this project, the Visible Human Project (VHP), the digital images currently available in the VHP database, ongoing research and development, and plans for the future of the VHP. PMID- 10386095 TI - Can virtue be taught? AB - Applying standards of virtue that define the "good doctor" in a complex and technologically sophisticated health care system is often challenging and sometimes confusing. What are the characteristics of a "good doctor," who wishes to live up to high ethical and professional standards but who also must live and work in a health care system in which moral ambiguity is pervasive? Medical educators are urgently faced with such questions as their schools try to equip students with the skills and capacities required of the virtuous physician. The author describes how Aristotelian concepts of virtue can be used to guide medical educators in defining and teaching virtue. He then discusses how such traits as the ability to tolerate moral differences and ambiguity, the ability to develop thoughtful individual moral positions, and the capacity to respect and understand various cultural traditions may be what might be considered virtues in today's health care system. A "good" doctor, then, would be someone who is thoughtful, fair-minded, respectful of differences, and committed to his or her professional values. PMID- 10386096 TI - Curriculum reform at the University of Otago Medical School. AB - The University of Otago Medical School, the older of the two medical schools in New Zealand, identified during the 1980s many of the same problems with its undergraduate curriculum as were reported in the United States, Canada, and the United Kingdom. An early, overly ambitious attempt to introduce a full problem based learning curriculum at Otago failed; however, many piecemeal changes that embodied some of the principles of problem-based learning were successfully implemented. Subsequently, as desire for more coordinated and substantive change grew, Otago's faculty used what they had learned from their first effort to successfully introduce a modular systems-based preclinical curriculum in 1997. The authors describe the features of the new curriculum and discuss two components (a systems-integration course and a large-scale program of computerized in-course testing) that are particularly innovative. The new curriculum is already achieving one of its main goals (increasing the perceived relevance of preclinical teaching) and other outcomes are being evaluated. PMID- 10386097 TI - Lessons for the new millennium from two ancient traditions. PMID- 10386098 TI - Streamlining the transfer of research tools. PMID- 10386099 TI - Graduate medical education training in clinical epidemiology, critical appraisal, and evidence-based medicine: a critical review of curricula. AB - PURPOSE: To systematically review the published literature on graduate medical education (GME) curricula in clinical epidemiology, critical appraisal, and evidence-based medicine (EBM). METHOD: The author searched the Medline and Educational Resources Information Center (ERIC) databases from 1973 through 1998, and also searched the references of the captured papers. The author reviewed all peer-reviewed reports of GME curricula (with or without effectiveness studies) in critical appraisal, clinical epidemiology, or evidence-based medicine, extracting objectives, formats, and evaluations (including effectiveness, process, and satisfaction). For effectiveness evaluations, he also identified the outcomes, outcome measures, methodologic characteristics, and results. RESULTS: The search produced 18 reports. The most common objective of the curricula described in the reports was improving critical skills; the most common format was resident directed small-group seminar. The most common outcome-evaluation measure was a multiple-choice examination. Only seven of the reports evaluated the curricula's effectiveness, and only four met a minimum methodologic standard of a pretest posttest controlled trial. The impacts on critical appraisal skills of the curricula in those four reports ranged from no effect to a 23% net absolute increase in test scores. CONCLUSION: These reports provide useful guides for medical educators, but many suffered from incomplete descriptions and inadequate evaluations of their curricula. The curricula themselves often focused on critical appraisal to the exclusion of other EBM skills and had limited effectiveness. In addition to increased methodologic rigor, future studies should focus on more meaningful outcome evaluations. Curricula should use residents' actual clinical experiences and teach EBM skills in real time in existing clinical and educational venues. PMID- 10386100 TI - Prevalences and correlates of ethnic harassment in the U.S. Women Physicians' Health Study. AB - PURPOSE: To describe lifetime prevalences and correlates of ethnic harassment in U.S. women physicians. METHOD: The authors analyzed responses to questions about ethnic harassment that appeared in the Women Physicians' Health Study, a 1993-94, nationally distributed survey of 4,501 female physicians. RESULTS: Of the responding physicians, 62% of blacks reported having experienced ethnic harassment, twice the rate of Asians and "others," three times that of Hispanics, and ten times that of whites. Twenty-five percent of black physicians reported experiencing harassment in at least three phases of their careers (before medical school, during medical school, during training, in practice), compared with 6% of "others," 2% of Hispanics and Asians, and less than 0.5% of whites. U.S.-born and foreign-born doctors reported similar rates of harassment before and during medical school, while foreign-born doctors reported significantly more harassment during training and practice. Reports of harassment during medical school were higher for blacks under 50 than for those over 50 (38% vs 10%, p = .0101). In white physicians, harassment was significantly associated with religion. For certain ethnic groups, control of work environment, dissatisfaction with profession, and stress at work and home were associated with reported harassment. CONCLUSIONS: The prevalences of ethnic harassment at various stages of medical training and practice are high, and not decreasing. This has serious potential ramifications for the medical profession's goal of a diverse physician workforce. PMID- 10386101 TI - Changing physicians' practices: the effect of individual feedback. AB - OBJECTIVE: To determine whether physicians who received feedback from six peers, six referring/referral physicians, six co-workers, and 25 patients about 55 aspects of their medical practices (e.g., able to reach doctor by phone after office hours) would make changes to their practices based on that feedback. METHOD: In an earlier study, 308 physicians were given feedback about 106 aspects of their practices in the form of mean Likert-scale ratings that (1) the peers made on 26 aspects; (2) the referring/referral physicians made on 23 aspects; (3) the co-workers made on 17 aspects; and (4) the patients made on 40 aspects. Three months later 255 of these physicians responded when asked to indicate whether they had contemplated or initiated changes, or whether no change had been necessary, regarding 31 practice aspects, each of which was a summary of one or more of 55 of the original 106 aspects on which they had received ratings. These 55 were considered the aspects most amenable to change over a short period. The physicians were also asked about the educational interventions that they felt would help them make changes. Multivariate analysis of variance was used to see whether the types of changes reported for the specific aspects of practice were associated with the feedback ratings received for those aspects. RESULTS: An examination of the responses showed that 83% of the 255 physicians reported having contemplated a change, and 66% reported having initiated a change for at least one aspect of practice. Changes were contemplated most frequently for aspects of practice associated with clinical skills and resource use. Changes were initiated most frequently for aspects of practice associated with communication with patients and support of patients. Physicians who contemplated or initiated changes had lower (i.e., more negative) mean ratings than did physicians who reported that no change was necessary, which suggests that the physicians did use their feedback ratings to decide about changes, although their qualitative comments indicated other sources as well. Printed material was chosen most often as a method of receiving continuing medical education related to making changes in the practice areas examined. PMID- 10386102 TI - What students value: learning outcomes in a required third-year ambulatory primary care clerkship. AB - PURPOSE: To determine learning outcomes from the students' perspective on the clinical portion of a third-year primary care ambulatory clerkship. METHOD: Over 18 months (December 1994 to June 1996), students at the Medical College of Wisconsin identified what they had learned during the clerkship in each of seven learning settings. Responses were transcribed and a coding dictionary developed. Response frequencies were compared by logistic regression analysis over time and between rural and urban sites. Course goals set by faculty were compared to learning outcomes reported by students. RESULTS: The authors coded 3,030 student outcomes into 48 categories. The top ten learning outcomes by frequency are reported. Logistic regression analysis revealed no significant difference by time of year or by rural versus urban clerkship experiences. Twenty-six of 29 original course goals were congruent with the student-generated outcomes. CONCLUSION: Basic professional knowledge, skills, and attitudes were the learning outcomes most valued throughout the year. Being alone with patients and working with their preceptors were the students' most valued learning settings. PMID- 10386103 TI - A comparison of outcomes for walk-in clinic patients who see interns and those who see staff physicians. AB - PURPOSE: To collect pilot data on the effect of interns' involvement on patient care outcomes. METHOD: Between January 1995 and August 1998, 750 patients at a walk-in clinic completed pre-visit questionnaires on symptom-related expectations and functional status. Three follow-up surveys (immediately after the visit, at two weeks, and at three months) assessed symptom outcomes, satisfaction, illness worry, and unmet expectations. Physicians were surveyed about their perceptions of the "difficulty" of each patient encounter. RESULTS: During the study period, 195 patients (26%) were seen by interns and 555 (74%) by staff physicians. The patient groups did not differ in illness worry, unmet expectations, or satisfaction immediately after the visit, at two weeks, or at three months. Patients seen by interns were more satisfied with the time they had spent with their clinicians (p = .007). Interns were more likely to experience their patient encounters as difficult. There was no difference in visit costs, subspecialty referrals, health utilization, or hospitalization rates. CONCLUSION: Patients who are seen by interns in an ambulatory clinic are similar to those who are seen by staff physicians in terms of post-visit satisfaction, residual expectations, symptom resolution, and functional status improvement. PMID- 10386104 TI - Educating new resident physicians in death notification. PMID- 10386105 TI - Results of the National Resident Matching Program for 1999. PMID- 10386106 TI - The changing landscape for clinical research. AB - The authors review the history of U.S. clinical research and identify the profound changes stemming from advancements in the biomedical sciences, the recent transformation in the organization and financing of health care delivery, and the increasing application of information technologies. They observe that the enterprise must reorganize to account for the changed landscape, but there is a lack of the data necessary to monitor change and determine the extent to which clinical research is successfully realigning and sustaining itself. The authors discuss the evolving definition, scope, and venues for clinical research, and review previous analyses of clinical research's difficulties and remedies proposed: shared responsibility in the financing of academic medicine, support by federal and private health insurers for routine costs of patient care in clinical trials, and strengthened collaboration between and among industry, academia, insurers, and government. The authors conclude by describing two major initiatives to foster clinical investigation in the new landscape. The first is the Clinical Research Summit Project, a convocation of representative stakeholders from the health care system with an interest in clinical research, whose charge will be to formulate a national agenda for clinical research that has the broad-based support of the stakeholders. Among the challenges of this undertaking are the needs to identify new and stable sources of support for clinical research infrastructure, assess the future workforce needs for clinical investigation, and devise new methods to ensure the continued vitality and account-ability of clinical research. The second is the Clinical Research Task Force, an initiative of the Association of American Medical Colleges (AAMC), which is already exploring and advising on how AAMC member organizations can best strengthen their capacity to support clinical research programs in the current scientific, health care delivery, and financial environment. PMID- 10386107 TI - Molecular medicine [is] for clinicians. PMID- 10386108 TI - Chest pain and abnormal T waves while at rest. PMID- 10386110 TI - What will the future hold for health care? PMID- 10386109 TI - A veteran with acute mental changes years after combat. AB - A 49-year-old man presented with a five-week history of worsening confusion, agitation, and bizarre behavior. According to his mother and sister, who live with him, he had inexplicably jumped out of bed one day and complained of injuring his back. The pain apparently resolved within several days. Two weeks later, again just after awakening, he had experienced a period of confusion, lasting about 15 min. The latest episode occurred three days previously and included vivid hallucinations--at various times, he seemed to believe that he was talking to his brother on the telephone, drinking a glass of water, emptying the refrigerator, jumping into a foxhole, and stomping on rattlesnakes. He was disoriented to time as well as environment. PMID- 10386111 TI - The human genome: information content and structure. AB - At a length of two meters and a width of two billionths of a meter, the genome amounts to a library whose code letters carry complete instructions for building and maintaining a human. Until recently, the code was modified only by mutation and natural selection. Today, however, we possess the technology to read and write DNA. The consequent knowledge is already changing medical practice. PMID- 10386112 TI - Manipulating angiogenesis against vascular disease. AB - Gene therapy to stimulate the growth of new blood vessels is proving to be an effective way of bypassing occluded arteries and reestablishing blood flow to ischemic tissues. It could eventually replace surgical revascularization and angioplasty, which are not only more invasive but also plagued with restenosis, problems limiting long-term management of coronary artery and peripheral vascular disease. PMID- 10386113 TI - Gout: beyond the stereotype. AB - Not all gout presents with involvement of the big toe, and not all gout patients are middle-aged men. Chronic gout may mimic rheumatoid arthritis; hyperuricemia may develop in postmenopausal women and in organ transplant recipients who are being treated with immunosuppressive agents. Both classic and nonclassic cases may benefit from new therapeutic agents. PMID- 10386114 TI - Medicinal herbs: a primer for primary care. AB - Used for centuries as folk remedies, herbs are enjoying a surge of public interest. Some empiric findings have been supported by formal research results. In order to advise patients about such preparations, physicians need to be aware of the indications, contraindications, drug interactions, and potential side effects. Eight popular herbs are reviewed. PMID- 10386115 TI - Primary care and pain medicine. A community solution to the public health problem of chronic pain. AB - The author emphasizes that pain is an important public health problem that demands attention. He discusses ineffective management and its causes, administrative and socioeconomic problems perpetuating poor care, problems in technology transfer, organizational models, specialists and subspecialists, and other topics. PMID- 10386117 TI - Postinjury neuropathic pain syndromes. AB - Pain is clearly one of the most daunting problems of modern medicine. Posttraumatic neuropathic pain syndromes are a major component of the clinical problem. Structural lesions affecting roots, nerves, the plexi, and central structures can be imaged noninvasively. The molecular biology of the intraneural cascades that cause sensitization of the central pain-projecting neurons of the dorsal horn and subsequent allodynia, hyperalgesia, and hyperpathia is a subject of intense inquiry. The role of the clinician in identifying and eliminating the source of the pain is crucial before the effects of excitotoxicity and central sensitization permanently alter the physiology of the central pain-projecting neurons and make treatment ineffectual. PMID- 10386116 TI - Central mechanisms in pain. AB - Nociceptive input into the central nervous system is not simply passively received but rather is subject to modulation through spinal cord neuroplasticity and descending influences from supraspinal sites activated by a variety of environmental signals, including the acute or persistent nociceptive input itself and behavioral and emotional stimuli. The significant role of NMDA receptors and production of NO. in central sensitization, hyperalgesia, and chronic pain has been demonstrated in numerous models of peripheral injury. It has been shown that persistent nociceptive input is also subject to centrifugal descending modulation through activation of both prominent facilitatory and masked inhibitory influences from supraspinal sites (e.g., RVM) likely involving a spino-bulbar spinal loop. These descending modulatory influences from the RVM appear to contribute selectively to hyperalgesia observed in uninjured tissue, distant from the site of insult (secondary hyperalgesia), and involve mechanisms similar to those found in the spinal cord (i.e., NMDA receptors and production of NO.). The significant role that modulatory influences in the central nervous system have in the development and maintenance of chronic pain and hyperalgesia clearly supports continued investigation into the physiologic mechanisms contributing to these events. PMID- 10386118 TI - Painful peripheral neuropathies. AB - This article reviews the current pathophysiology of painful peripheral neuropathies, their differential diagnosis, and management. PMID- 10386120 TI - Headache disorders. AB - Head, neck, and facial pain disorders possess characteristic features that, in some ways, distinguish them from other painful disorders. Generally speaking, the headache disorders can be reconciled within the same model of assessment as that of other painful conditions. PMID- 10386119 TI - Low back pain. Evaluation and management in the primary care setting. AB - The primary care physician plays a major role in the identification of low back pain and the entry of the patient into the health care system. Acute low back pain remits within a short period of time in most patients, and major diagnostic studies are not required. If the pain persists beyond the treatment parameters of the primary care physician, consultation is necessary. A basic component of the initial evaluation is the identification of myofascial syndromes that mimic so called root syndromes. Further, low back pain in the population at large is not usually a surgical problem, and the chances of there being significant pathology requiring surgical or other forms of intervention may be less than 1% of those affected. When the initial attempts at treatment fail, the patient should be referred to a multidisciplinary comprehensive pain center so as to avoid or limit chronicity, the earlier, the better. Practitioners should feel comfortable in asking the centers to which they make a referral for outcome data. If these are not available, the choice should be made elsewhere. Low back pain per se is in the majority not a neurologic problem, an orthopedic problem, or a neurosurgical problem, so that consultation with these groups, unless there are strong suspicions otherwise, has limited value. The criteria for selection and referral of patients to multidisciplinary pain centers have been presented, including specific considerations for the geriatric age group. The overwhelming cost of low back pain to the economy can be decreased along with suffering and the adverse impact that pain has on all social strata. PMID- 10386122 TI - Cancer pain management. AB - Pain control is vital to the quality of life of patients with cancer. From pain assessment and pharmacologic management to interventional management, special features of cancer pain are outlined in this article. Many evidence-based national and societal guidelines were developed during the past decade on cancer pain management. Proper and adequate management of cancer pain is strongly recommended by all the clinical practice guidelines. PMID- 10386121 TI - Medically unexplained chronic orofacial pain. Temporomandibular pain and dysfunction syndrome, orofacial phantom pain, burning mouth syndrome, and trigeminal neuralgia. AB - Orofacial pain syndromes pose a dilemma for physicians. Even when the patient is referred, quality medical care requires that the physician be acquainted with current evidence-based practice. Such practice may be radically different from the traditional view. This article reviews the differential diagnosis and treatment of the most common medically unexplained orofacial syndromes. PMID- 10386123 TI - Approaches to treatment decisions for psychiatric comorbidity in the management of the chronic pain patient. AB - A number of different types of comorbidities have been described within psychiatric patients. These comorbidity types are reviewed and their application to the chronic pain population is discussed. These various types of comorbidities are then utilized to generate an approach for treatment decisions in the management of the chronic pain patient. PMID- 10386124 TI - Opioid use in the management of chronic pain. AB - Opioids are a necessary and effective component of the management of chronic non cancer-related pain in some patients. Careful structuring, monitoring, and documentation of care are important, but the therapeutic use of opioids is uncomplicated in the majority of patients using opioids and is gratifying for both the patient and the treating physician when it results in significant reduction in pain, improvement in level of function, and a higher quality of life. Opioid therapy is most often successful when combined with other pharmacologic and nonpharmacologic interventions as indicated by the type of pain and the context in which it occurs. PMID- 10386125 TI - Interventional pain management. Appropriate when less invasive therapies fail to provide adequate analgesia. AB - Unrelieved chronic pain is costly to patients and society. Noninvasive and less costly therapies should be used before more invasive and more costly therapies. Therapies for pain control should be used according to a pain treatment continuum. Nerve-blocking techniques, neurolytic techniques, and implantable neuromodulatory technologies, such as SCC and spinal delivery of analgesics, are cost-effective when less invasive therapies fail to provide adequate analgesia. PMID- 10386126 TI - Roadblocks to effective pain treatment. AB - Managed care "backlash" rhetoric to the contrary, roadblocks to effective pain treatment occur both intrinsic and extrinsic to the healthcare system. Pain medicine, an emerging, formally recognized specialty, and the special population of patients which it serves, experience additional discreet barriers. Chief among these is a lack of clear identity and recognition of the specialty and the disenfranchisement of many of the patients it serves in the American healthcare system. Special problems within various healthcare financing environments is discussed. PMID- 10386127 TI - Treatment planning in pain medicine. Integrating medical, physical, and behavioral therapies. AB - This article addresses a systematic approach to the treatment of chronic pain. The first section presents a biopsychosocial model of pain. The second section presents an application of the biopsychosocial approach to the clinical assessment and management of clinical cases with chronic pain. PMID- 10386128 TI - [The tuberculosis situation in the world in 1996]. PMID- 10386130 TI - [The results of using a simplified questionnaire for determining the prevalence of bronchial asthma]. AB - The aim of the study was to assign the asthma prevalence in a Romanian region, by using a simplified questionnaire, easy to fill in by every subject without specialized help. The study included 508 subjects, 280 women (55.1%) and 238 men (44.9%). The subjects proceeded from different industrial areas (cement factory, rubber processing factory, fur and leather manufacture) and from a village (214 subjects). 203 subjects, having symptoms compatible with asthma, performed lung function tests with a Flow Screen Jaeger device, determining VC, FEV1, FEV1% VC, MEF50, MEF25, and pharmacodynamic test if needed (bronchoconstrictor or bronchodilator). RESULTS: about 40% of the subjects mentioned the wheezing, among them 18% wheezing and dyspnea. "Asthmatic bronchitis" is present at 14.8% of the subjects, including 4.5% patients with bronchial asthma diagnosed by a physician (underdiagnosis). The asthma symptoms are more frequent in the factories with exposure to inhaled allergens and air pollution. The most discriminative symptom association for asthma was: wheezing and breathlessness and a history of dyspnea in the last year. These symptoms suggest that the prevalence of asthma could be 10.43%. The positive bronchodilator or bronchoconstrictor function test associated with wheezing, present at 7.48% of the population, seems to better evaluate the prevalence of asthma, which approaches other data already published. The questionnaire we used proved that it can select with acceptable accuracy the individuals to be further investigated with lung function tests. PMID- 10386129 TI - [A survey of recommended measures in COPD (II)]. AB - The authors aimed to test the doctors' opinion-general practitioners (Gen), internal medicine specialists (Int) and pulmonologists (PNF) about the therapeutic approach in COPD. 300 questionnaires including 53 questions were mailed; 103 filled-in questionnaires were recovered (33.34%). Most of the doctors consider necessary the initial diagnosis in hospital. The Gen send the patients mostly to the Int and less to the PNF. Most doctors accept that the treatment should be started in pulmonology clinics. All the doctors prescribe teophillins; the beta-agonists are prescribed twice as much by the PNF than Int, and very little by Gen. The oral corticosteroids are used by half of the doctors; the inhaled corticoids are prescribed seldom and only by PNF. The i.v. corticoids are used by half of Int. All the doctors recommend smoking cessation. Only the PNF drag the attention upon the bacterial infections. The kinesitherapy is seldom recommended, even by PNF. PMID- 10386131 TI - The relationship between serum ACE activity and total IgE levels in patients with bronchial asthma. AB - Pulmonary endothelium takes part in the metabolization of some products such as prostaglandins, norepinefrine, serotonin, bradikinin and angiotensin I. Angiotensin I is the substrate for Angiotensin Converting Enzyme (ACE). It is known that greatest production site of ACE is the pulmonary endothelium. A lot of studies have been done for determining the levels of ACE in various lung diseases. It has been observed that serum ACE activity is increased in granulomatous diseases such as sarcoidosis and berylliosis. In patients with bronchial carcinoma, serum ACE activity is found to be decreased. There are some papers about the significant changes in serum ACE activities in asthmatic patients. We determined the activity of ACE and total IgE levels at the serum of 40 asthmatic patients and 20 healthy subjects. Among 40 patients 20 of them were mild asthmatic and they developed symptoms only during the attacks. The remainder 20 patients were chronic asthmatics and all the time they had the symptoms of dyspnea, wheezing and coughing. Serum ACE activity was found 25.5 +/- 11.77 U in the control group, 22.8 +/- 8.04 U in mild asthmatic group and 16.6 +/- 6.13 U in chronic asthmatic group. When compared with control group serum ACE levels found to be significantly decreased in chronic asthmatic group. Also a weak but significant correlation was found between serum total IgE levels and ACE activity in the chronic asthmatic group. These findings suggest that there is a relation between ACE and Total IgE production. PMID- 10386132 TI - [The characteristics of a case of Churg-Strauss syndrome (CSS)]. AB - The authors present the case of a 35-years-old female with severe bronchial asthma non-responding to treatment, mobile infiltrative pulmonary images on chest X-ray considered and treated first as pulmonary tuberculosis smear-negative, maculo-papular skin lesions on the limbs and a high percentage of eosinophils in peripheral blood. The diagnosis was settled by histologic examination of the skin lesions. The evolution was very good with high doses of oral corticosteroids. PMID- 10386133 TI - [Diagnostic difficulties--salvage diagnostic thoracotomies]. AB - The authors are presenting the difficulties of diagnosis, in clearing up the etiology in two cases, where the acceptance of the exploring thoracotomy, influenced the prognostic and permitted to settle up the diagnosis. We are passing through, at this occasion, also the indications of this kind of operation. PMID- 10386134 TI - [The electrophoretic study of serum proteins in pneumophthisiology]. PMID- 10386135 TI - [Oxygen therapy and ventilation or the long road from discovery to prescription]. PMID- 10386136 TI - [Antitubercular drugs]. PMID- 10386137 TI - [Tuberculosis at the Global Congress on Lung Health and the 1997 annual meeting of the International Union against Tuberculosis and Lung Disease (UICTMR), Paris, France, 1-4 October 1997]. PMID- 10386140 TI - [Youth and the temptation to smoke]. PMID- 10386143 TI - [The evolution of the incidence of tuberculosis in Romania in the 1st 6 months of 1998]. PMID- 10386144 TI - [The prevalence of respiratory symptoms, bronchial asthma and chronic bronchitis (simple and obstructive) in a representative sample of the adult rural population]. AB - A representative sample of an adult population living in a rural area (1001 adult subjects aged 18 and above) was submitted to the ECHRS standardised questionnaire (administrated by interview) and performed lung function tests (ventilatory). The most frequent respiratory symptoms were chronic cough (especially in males and smokers) and intermittent dyspnoea (in females more prevalent than in males). All symptoms showed a higher frequency after 50 years. Obstructive ventilatory impairment was found in 12.29% of the subjects, more frequently in males, ex smokers and above 50 years of age. The prevalence of asthma bronchiale was 4.09%; the subjects over 40 years of age, the non-smokers and the females showed higher figures. Chronic simple bronchitis was found in 5.69% of examined subjects, especially in males and smokers. The prevalence of chronic obstructive bronchitis was as high as 2.29%, more prevalent in males, smokers, over 40 years. The prevalence rates found in this study are higher than those observed in a rural population living in an unpolluted area: this fact may be ascribed to the noxious effect of the air pollution due to the chemical industrial plants situated in the vicinity of the studied population. PMID- 10386145 TI - [The bacterial spectrum in bronchopulmonary infections]. AB - This study aimed to establish the bacterial profile of bronchopulmonary infections confirmed by the cytobacteriological examination of sputum, in order to find therapeutical guidelines for empirical treatment. We included in the study 408 patients with clinical signs of bronchopulmonary infection (cough and mucopurulent sputum, fever) among which 294 hospitalised patients (5.5% of the 5280 admitted in 1997) and 114 outpatients. The sputum samples collected respecting the decontamination methods were examined cytobacteriologically (smear, culture and antibiogram). The spectrum of isolated bacteria was the following: H. influenzae--198 cases, anaerobes--54, Kl. pneumoniae--53, Ps. aeruginosa--50, S. pneumoniae--17. Analyzing the diseases for which the bacterial examination was performed, we found the following distribution: COPD--66, bacterial infections in TB patients--61, chronic suppurations--33, bronchiectasis -37, pulmonary abscess--24. We noticed the high frequency of H. influenzae and important numbers of anaerobes, Kl. pneumoniae and Ps. aeruginosa especially in COPD patients and patients with chronic suppurations. We performed antibiograms for establishing the sensitivity of bacterial samples (S. aureus, Kl. pneumoniae, Ps. aeruginosa, beta-lactamase-positive H. influenzae). Most of them were multidrug-resistant. CONCLUSIONS: 1. The cytobacteriological study of sputum may be useful for choosing the right treatment, especially for the patients with multiple antibiotic treatments and infected with multidrug resistant bacteria; 2. Knowing the bacterial spectrum in certain respiratory diseases allows the choice of empirical treatment for bronchopulmonary infections in uncomplicated cases. PMID- 10386147 TI - [The incidence and prevalence of chronic pulmonary tuberculosis in Galati County]. AB - In Galati County, the tuberculosis is augmenting constantly, the incidence increased from 46.6/100,000 in 1985 to 64.5/100,000 in 1990 and 112.2/100,000 in 1997. The incidence (new and readmitted cases) increased by 140% (2.4 times) in 1997 compared to 1985, the level in 1997 being higher than the mean level of the whole country. About 80% of the patients eliminated mycobacteria at the time of diagnosis, and 80% of these had positive smears. Thus, the periodic prevalence of the smear positive increased, and the children's risk also. An important factor for supplying the secondary pool with new infected individuals are the chronic and hyper-chronic patients (group IB and IC). In the last 3 years 281 chronic patients were registered, 218 in group IB and 63 in IC. The main cause for the failure of treatment are incomplete, irregular therapy and uncooperative patients. Chronic patients are also the source of infection with multidrug resistant germs. The high prevalence of chronic patients justifies also the increase in TB mortality in the last 7 years: 4.2/100,000 in 1990 and 12.9/100,000 in 1996. Only by increasing the curing rate and reducing the therapeutic failure can the chronic TB problem be solved. PMID- 10386146 TI - [The distribution of tuberculin allergy among students in their 5th and 6th years at the University of Medicine and Pharmacy Iasi]. AB - The authors studied the distribution of tuberculin allergy in a group of high TB risk, the students in last years of study of the Iasi University of Medicine. They are already BCG vaccinated. An infection prevalence of 18.6% was found- reactions over 18 mm, also a reaction of 10-17 mm was found at 62.2% of the students having 2-3 BCG scars. In the time of the present national BCG vaccination program there is need for new criteria for separating the two types of allergy: post BCG and post infection. PMID- 10386148 TI - [The clinico-epidemiological aspects of tuberculosis occurring in medical personnel in pneumophthisiology units in Bucharest (1992-1996)]. AB - The authors studied 38 TB cases diagnosed between 1992 and 1996 among the 1092 employees (medical and nonmedical) of the pneumophthisiology departments in Bucharest (hospitals and ambulatory units). Most cases had pulmonary involvement (76.3%), 85.7% were bacteriologically tested for bK. Among them, 66.7% were confirmed. The therapeutic results evaluated after 1 year, showed just 1 failure. The medium annual risk of tuberculosis at the PF personnel is corresponding to an incidence of 700/100,000 (6.36 times higher than TB incidence in general population). This conclusions calls for emergency measures for protecting and compensating this personnel. PMID- 10386149 TI - [Fiber bronchoscopy in the diagnosis of bronchial asthma]. PMID- 10386150 TI - [Respiratory involvement in neutropenic patients]. PMID- 10386151 TI - [Pulmonary aspergilloma successfully operated on in a patient with the pentalogy of Fallot]. AB - Fallot disease occurs in 10% of congenital heart diseases. This case showed a clinical association between Fallot disease and a pulmonary aspergilloma. A 44 years male, with a history of Fallot disease (diagnosed in childhood) and a normal life, was admitted for repeated hemoptysis during last three months. Clinical examination revealed signs of Fallot disease: cyanosis, finger clubbing. CT scan and chest X-ray revealed a cavitary image in the right upper lobe. The CT image revealed a typical aspect of cavitary aspergilloma. The patient was referred to the thoracic surgeon, a right upper lobectomy was performed, which confirmed the presence of aspergilloma. Postoperative evolution was good. PMID- 10386152 TI - [A case of tracheobronchopathia chondroplastica]. AB - A new case of tracheobronchopathia osteochondroplastica is reported, demanding for the diagnosis besides the radio-clinical investigations, also the pathologic proof. The etiology is unknown. PMID- 10386153 TI - [Preoperative evaluation in thoracic surgery]. PMID- 10386154 TI - [Rapid diagnosis in sero-hemorrhagic pleurisy]. PMID- 10386155 TI - [Little "tricks" in the practice of isolated fiber bronchoscopy]. PMID- 10386156 TI - [A "pseudodictionary" of bronchial asthma]. PMID- 10386157 TI - [Acquaintance with the National Antituberculosis Program 1997-2000?]. PMID- 10386158 TI - Predictable new developments in occupational health activities. AB - Current environmental problems are discussed in reference to the Japan Medical Association's Committee report on Environmental Health. Attentions are particularly placed on transition in environmental problems, trends of global environmental changes, chemicals like dioxines, exogenous endocrine disrupting chemicals, and role of medical associations coping with health problems accompanied with environmental changes. PMID- 10386159 TI - Platelet membrane protein CD36. AB - CD36 is one of the major glycoproteins of platelets and known as GPIV. Besides platelets, CD36 is distributed in megakaryocytes, monocytes, capillary endothelium and mammary epithelial cells. In vitro analyses, CD36 is reported to act as receptors to a variety of ligands including collagen, thrombospondin, malaria-infected erythrocytes and oxidized LDL. However, it remains unclear to which of these functions CD36 is critical in vivo. CD36-deficient individuals can be the key to answer this question. In calcium-deficient state, CD36-deficient platelets exhibited a delay and decline of irreversible aggregation on agonist stimulation. Irreversible aggregation of platelets depends on intake of arachidonic acid once-secreted from platelets and production of its metabolite Eps/TxA2. The calcium influx in response to U46619 (TxA2 analogue) of CD36 deficient platelets was not different from normal platelets in the presence of indomethacin and ETYA. Defective aggregation of CD36-deficient platelets in calcium-deficient state seemed to be derived from defective intake of arachidonic acid. This assumption was verified by our results that inhibitory effect of arachidonic acid in aggregation depended on the presence of platelet CD36. Intake of arachidonic acid through CD36 may have an effect in low concentration state of arachidonic acid. The CD36 deficiency is present in several % in Japanese and approximately 0.3% in Caucasians and is divided in type I (deficient in platelets and monocytes) and type II (deficient only in platelets). Analyses of CD36 cDNA revealed that codon 90 (proline/serine) was critical as to the surface expression of CD36 protein. By analyses of CD36 genomic DNA, the CD36 gene could be classified; 1) serine90 type that was not translated as CD36 protein, 2) proline90 type that was not transcribed to mRNA, 3) proline90 type that was transcribed only in monocytes and not in platelets, 4) proline90 type that was transcribed in platelets but in very small amounts and 5) wild type proline 90. The results of family studies were consistent with the assumption described above. PMID- 10386160 TI - Non-surgical treatments for chronic pancreatitis. AB - Non-surgical, less invasive treatment has been required for chronic pancreatitis. We attempted endoscopic approach and extracorporeal shock wave lithotripsy (ESWL) for the treatment of 54 patients with chronic pancreatitis. Endoscopic procedures, such as endoscopic pancreatic sphincterotomy, cystenterostomy and endoprosthesis, were performed in selected 11 patients with pancreatic stones, protein plugs, pseudocysts and main pancreatic duct stenosis. Successful results were obtained in all 11 patients. ESWL was used for the treatment of 43 patients with pancreatic stones. Combined with ESWL, endoscopic treatments were also used in 7 patients. Stone disintegration was achieved in 36/43 patients (83.8%), and complete clearance of main pancreatic duct was obtained in 19 (44.1%). Pain relief was observed in 27/29 (93%). Improvement of exocrine pancreatic function, evaluated by PFD test, was recognized in 10/19 (52.6%). ESWL is a safe and effective treatment for pancreatolithiasis. Endoscopic approach is, when it will be carefully performed, a less invasive and useful treatment for main pancreatic duct stenosis or pancreatic pseudocyst. PMID- 10386161 TI - The reconsideration of natural history of echinococcosis at Rebun Island. AB - It has been believed that the outbreak of echinococcosis at Rebun Island had ceased by 1970. The first patient was diagnosed in 1936 and 131 patients have been authorized as echinococcosis so far. The conference of measures against the outbreak had been organized in 1948 and started to eradicate Echinococcus multilocularis from the Island. Medical examination to detect the patients and the capture and autopsy of dogs and cats had been carried out hard till 1970. At that time, foxes imported from Simusiru Island in the middle Kuriles during the years 1924 to 1926 had already disappeared and it has seemed to be sure that stray dogs and cats might carry E. multilocularis and excrete infectious eggs in stead of foxes. Since we have had no real data concerning the natural history of patients with echinococcosis without any treatments, it can not be recognized the time of infection and the role of dogs or cats on the spread of echinococcosis at Rebun Island. From the new data, it is concluded that the active life cycle of E. multilocularis between foxes and vole might be closed by 1940, since the last patient infected with E. multilocularis was born in 1940 and died in 1945. Furthermore, it is estimated that more than 200 patients (3 to 4% of people at the island) might die from echinococcosis, because of the fact of the unusual increase of mortality of liver disorders and oldness observed during the years of 1940 to 1960. 81 patients with the high possibility of echinococcosis detected from 1937 to 1963 can be added to 131 authorized patients. Surprisingly, it is noticed that the standard deviations of ages of death of 94 patients born in Meiji era (1880-1912) and 59 in Taisho and Showa eras (1912-1940) are 63.16 +/- 11.68, and 34.32 +/- 11.87, respectively. It means that both old and young people might be infected simultaneously but for the long period. There was no difference between the susceptibility of young and old men to E. multilocularis. The numbers of male patients died were more than those of female patients at the ages from 30s to 60s while the number of female died was predominant after 70s. All 13 familial cases of echinococcosis represent that men might bring the infectious eggs into their houses and died earlier by the infection. Thus, the sexual difference might be due to the life style of men who preferred hunting beside fishing. From these results, it is conceivable that the heavy infection of E. multilocularis excreted from foxes might occur from 1925 to 1940 and the peak of the death might be formed during 1940 to 1965. The eradication of foxes might be done by poachers after 1935 and the adaptation of E. multilocularis from fox to dog or cat might not occur readily at Rebun Island. PMID- 10386162 TI - The role of gamma-aminobutyric acid (GABA)-benzodiazepine neurotransmission in an animal model of methamphetamine-induced psychosis. AB - Repeated administration of amphetamine or methamphetamine (MA) results in an augmentation of its locomotor-activating effects, which is a phenomenon known as behavioral sensitization. In humans, chronic use of the drug elicits a progressive augmentation in paranoid symptoms that closely resemble schizophrenia. Behavioral sensitization has some common properties with other forms of neural plasticity such as kindling, learning and long-term potentiation (LTP). The author examined in the present study whether behavioral sensitization could be blocked by GABA-benzodiazepine agonists, known to inhibit kindling, learning as well as LTP. Rats (Male Wistar-King rats) treated with MA (1 mg/kg, s.c.) for 10 days displayed significantly enhanced motor activity when tested with MA (1 mg/kg) after a 7-8 day withdrawal period indicating the acquisition of behavioral sensitization. Treatment with clonazepam (CZP) (0.5 and 2.0 mg/kg), a GABA-benzodiazepine agonist, prior to MA administration prevented the acquisition of sensitization. In contrast, treatment with flumazenil (Flu) (10 mg/kg), a GABA benzodiazepine antagonist, prior to MA administration did not affect the acquisition of sensitization. And treatment with Flu prior to CZP administration suppressed the inhibitory effect of CZP. CZP had no effect on the expression of sensitization in the sensitized rats, when given prior to the MA readministration. These results suggest that stimulation of GABA-benzodiazepine receptors plays a role in the acquisition but not in the expression of behavioral sensitization. PMID- 10386163 TI - Analysis of matrix metalloproteinases and related tissue inhibitors in cystic fluids of ovarian tumors. AB - Proteolytic activity of cystic neoplasms of the ovary appears to play a role in destruction of the extracellular matrix and tumor invasion. The purpose of this study was to determine whether the enzymatic activities reflect the degrees of tumor malignancy. The author examined the activity and quantity of matrix metalloproteinases (MMPs) and tissue inhibitors of MMP (TIMPs) in cystic fluids of both benign and malignant epithelial ovarian tumors. The concentration of MMP 9 was statistically higher in mucinous carcinomas (p < 0.05) than in benign ones. TIMP-1, which combines with MMP-9, was also higher (p < 0.05) in malignancies than in benign ones. The ratios of MMP-9/MMP-2 and the concentrations of activated forms of MMPs well associated with the degrees of malignancy, while the mol ratios of TIMP-1/MMP-9 and TIMP-2/MMP-2 inversely correlated. Expressions of MMP-3 and/or trypsin in the fluids were frequently associated with activation of MMP-7 and MMP-9. These observations support the concept that the imbalance of TIMPs/MMPs and the activation of MMPs correlate with the biological malignancy of ovarian cystic tumors. PMID- 10386164 TI - Analysis of hematopoietic cell specific protein, M-DOCK. AB - Full length cDNA of M-DOCK was isolated at Kazusa DNA Institute and its sequence has been deposited as KIAA0209. The cDNA sequence of M-DOCK contains a 5490 bp open reading frame that encodes a 1830-aa protein with a calculated molecular mass of 212 kDa. The amino acid sequence of M-DOCK has 62.3% identity with DOCK180, excluding the carboxyl terminal variable regions. DOCK180, which was originally identified as a major protein bound to Crk oncogene product, is an archetype of a family of proteins including Ced-5 of C. elegans and Mbc of D. melanogaster. DOCK180 is localized at focal adhesions and regulates cell morphology through the activation of Rac. Expression of DOCK180 is ubiquitous except in hematopoietic cells. Here, the author reports on the characterization of M-DOCK protein, which is closely related to DOCK180 but expressed only in the hematopoietic cells. We immunized rabbits with bacterially-expressed GST-M-DOCK protein, and obtained an anti-serum specific to M-DOCK. Tissue distribution of M DOCK was examined by Northern blotting and Western blotting. M-DOCK was expressed abundantly in peripheral blood leukocytes. M-DOCK expression was detected also in the cell lines derived from T-cells, B-cells, and monocytes, but never in the adherent cells. Most of the lymphocytes and macrophages in human organs expressed M-DOCK diffusely in the cytoplasm, which was analyzed by immunohistochemistry using an anti-M-DOCK antibody. We previously reported that membrane-targeted form of DOCK180 induced spreading of NIH 3T3 cells. By contrast, overexpression of M DOCK rounded up flat NRK cells. This suggests that M-DOCK may regulate cell detachment of adherent cells. These results demonstrate that M-DOCK is a new member of DOCK180-family proteins and regulates cell shape in suspension cells. PMID- 10386165 TI - Long telomeres and well preserved proliferative vigor in cells from centenarians: a contribution to longevity? PMID- 10386166 TI - Postoperative experiences of pain and distress in elderly patients. An explorative study. AB - The aim of this study was to explore postoperative experiences of pain and distress in elderly patients, as well as interventions aimed at reducing these conditions, on three occasions. The study group was composed of 100 patients who had undergone elective surgery in two orthopedic and two general surgical units. Of the 50 patients in the orthopedic units, 26 had undergone hip arthroplasty and 24 knee arthroplasty: of the 50 patients in the general surgical units, 23 had had breast cancer surgery, and 27 abdominal surgery. The patients were interviewed, using a structured interview format, on three occasions; at the ward on the first and second day after surgery, and by telephone about ten days after discharge from hospital. Within both the sensory and the emotional dimensions, logistic regression analyses showed that the dependent variables of pain and distress were significantly related above all to type of surgery and sense of coherence (SOC). In a cluster analysis, three meaningful clusters of patients were obtained. The patients in the different profiles showed variations in their experiences of pain and distress. The 12 patients with the least favorable scores had weaker SOC than the patients in the other profiles. It is concluded that type of surgery and psychological factors influenced patients' experiences of pain and distress after undergoing surgery. These experiences should be reduced by identifying risk patients, and improving assistance and support in the nursing ward, and also when patients have returned home. PMID- 10386167 TI - Targeting and quality of nursing home care. A five-nation study. AB - The objective of this study was to demonstrate that appropriate targeting and quality monitoring of institutional care of the elderly is possible using person based information on residents of nursing homes. This cross-sectional study used Minimum Data Set (MDS) assessments of nursing home residents in 6 US states, Copenhagen, Reykjavik, and selected locations in Italy and Japan. The outcome measures were life expectancy at age 65, population over 65, percentage over 65's in nursing homes, and clinical characteristics of nursing home residents from a multinational database of RAI/MDS assessments. We found that Japan has the highest life expectancy, and the second lowest expenditure on health care. The United States has the highest expenditure on health care and intermediate life expectancy. Italy has the highest proportion of population over 65 and the lowest proportion of over 65's in nursing homes. Iceland, a relatively young country, has the highest proportion of over 65's in nursing homes. Residents in Italy and the United States had the most severe physical, cognitive and clinical characteristics, those in Iceland the least. There was wide variation in markers of quality of care, with no country either uniformly good or bad across multiple measures. In conclusion, headline statistics comparing nations' percentage of Gross Domestic Product (GDP) spent on health care, age structure of the population, percentage of over 65's in nursing homes and clinical characteristics bear no consistent relationship. Local policy and practice also affect quality of care. Standardized assessment enables comparisons at local, national and international levels making possible further research on targeting and the appropriate use of institutional care, thus permitting a range of efficiency measures to be developed to inform policy. PMID- 10386168 TI - Coexistence of lowered mood and cognitive impairment of elderly people in five birth cohorts. AB - The prevalences of lowered mood and cognitive impairment, and their combination were investigated in 1993 random subjects of five birth cohorts (at age of 65, 70, 75, 80 and 85 years). The frequency of a high Zung-score (> 45), indicating depressive symptoms, in the five age groups was 11%, 13%, 20%, 16%, and 36%, respectively. The corresponding figures for a low MMSE-score (Mini Mental State Examination < 24) were 11%, 9%, 25%, 46%, and 60%; the respective frequencies of subjects fulfilling both criteria simultaneously were 2%, 3%, 8%, 12% and 24%, respectively. Overall, about 30% of the subjects with a low MMSE-score had a high Zung-score. However, more than half of the old subjects (over 75 years) with a high Zung-score also had low MMSE-scores. The data indicate that the combination of impaired cognition and lowered mood doubles in frequency by five-year intervals after the age of 70 years in the general aged population, and that this condition is present in one of four subjects at the age of 85 years. PMID- 10386169 TI - Mental health in senior citizens in the metropolitan zone of Guadalajara. AB - Using Goldberg's General Health Questionnaire (GHQ) to identify potential cases of mental disorders, we screened 246 randomly selected persons among the 116,616 people older than 65 in the metropolitan zone of Guadalajara; 86 subjects (35%) qualified as "cases"; this figure is much higher than that reported (20.8%) in the adult population in a previous study. Among the cases, 69% were female, 66% were widowed, and 50% were divorced; 44% had not finished gradeschool, 42% had no scholastic education, 54% were unemployed, and 40% worked at home. Factors associated with anxiety and severe depression, sleep disorders, psychosomatic symptoms, and problems in interpersonal relations were studied. PMID- 10386170 TI - Reduced water intake but normal response to acute water deprivation in elderly rhesus monkeys. AB - The ability to compensate for acute water deprivation was studied in young adult (YA, 7-9 years), middle aged (MA, 13-17 years), and older adult (OA, 20-36 years) rhesus monkeys of both sexes (N = 6/group). Water intake and urine volume were measured during three 7-day trials: 3 days of baseline measurement, 1 day of deprivation and 3 days of compensation. OA drank less during baseline (380 +/- 63 mL/day) than MA (679 +/- 92 mL/day, p < 0.05) or YA (750 +/- 128 mL/day, p < 0.01). All groups drank more following deprivation than at baseline and the OA drank significantly less than the younger groups (both p < 0.01), but the increase above baseline did not differ among groups when expressed as a cumulative percentage of baseline (89% for OA; 77% for MA; 83% for YA). Urine volume of all groups decreased by similar percentages on the day of deprivation (56% overall) and this reduction represented a similar proportion (58% overall) of baseline water intake. Urine concentration increased significantly during deprivation (p < 0.05) and returned to baseline values during compensation with no differences among age groups. OA water balance appears to have been maintained at lower levels of intake and excretion. In conclusion, responses to acute hydrational challenges in the elderly should be interpreted in the context of customary fluid intake. PMID- 10386171 TI - Contractile properties and protein isoforms of single skeletal muscle fibers from 12- and 30-month-old Fischer 344 brown Norway F1 hybrid rats. AB - This study characterizes single skeletal muscle fiber contractile properties and myosin heavy chain (MHC) isoform compositions from the soleus (SOL) and deep portion of the lateral head of the gastrocnemius (RG) muscles of 12- and 30-month old Fischer 344 Brown Norway F1 hybrid rats (FBN F1). Thirty months of age is approximately the age of 50% survival for the FBN F1 rat. For type I MHC individual fibers from the SOL of 30-month-old animals, the diameter was 88 +/- 2 microns, peak active force was 4.4 +/- 0.2 x 10(-4) N, peak specific tension (P0) was 76 +/- 5 kN/m2, and maximal unloaded shortening velocity (V0) was 0.98 +/- 0.09 fl/s. The type I MHC fibers from the SOL of 12-month-old animals had similar properties with the exception of P0 which was 92 +/- 4 kN/m2 and V0 which was 1.65 +/- 0.12 fl/s. Contractile properties of the RG MHC type I fibers were not significantly different from MHC type I fibers from the SOL in both age groups. The V0 of the RG type IIa MHC fibers from the 12-month animals (4.05 +/- 0.36 fl/s) and 30-month animals (3.55 +/- 0.41 fl/s) and of fibers co-expressing type I MHC and type IIa MHC from 12-month animals (4.21 +/- 0.55 fl/s) and 30-month animals (2.22 +/- 0.27 fl/s) were significantly faster than that of MHC type I fibers of the respective age group. In conclusion, skeletal fibers from the 12- and 30-month-old FBN F1 rats demonstrate fiber-type specific properties with a close relationship between the MHC isoform composition and the V0. PMID- 10386172 TI - Changes in the composition of human unstimulated whole saliva with age. AB - The aim of this study was to evaluate changes in the concentration of certain components of human unstimulated whole saliva during aging, in order to better understand the role played by aging in oral health. In particular, we studied total protein concentration, alpha-amylase activity, sialic acid content and calcium and phosphorus concentrations in 100 healthy subjects of both genders, aged between 10 and 80 years, who were subdivided into four groups according to their age: 10-25 years, 26-40 years, 41-65 years, and 66-80 years. Other than sialic acid, the concentrations of the components studied were not affected by age. There was a significant negative correlation between sialic acid content and age. Our data indicate the presence of a decreased submandibular/sublingual function with aging, thus suggesting the possibility of a concomitant reduction in the modulating action of unstimulated whole saliva on the oral flora. PMID- 10386173 TI - Evaluation of complex activities in daily living of elderly Japanese with visual impairment. AB - This study was conducted to determine whether elderly subjects with visual impairment differ in the performance of complex activities in daily living from those without visual impairment. The study subjects were residents in two homes for the aged in Japan, and consisted of 37 elderly people with visual impairment, and 42 elderly people, serving as controls; ages ranged from 64 to 95 years. Complex activities of the subjects were ascertained by interview using a 46-item questionnaire. The visually impaired elderly had lower performance levels for telephone use (p = 0.007), shopping (p = 0.049), cleaning up one's room (p = 0.001), and utilization of medical facilities (p = 0.001) in instrumental ADL (IADL); for interest in TV or radio (p = 0.004) and religious faith (p = 0.042) in "enriching activities"; and for visiting behaviors (p < 0.05) in "social role". The performances of complex activities by the elderly with visual impairment were diminished in specific categories, but not overall, and this may be attributable to poor mobility and/or more passive attitudes in their daily activities. PMID- 10386174 TI - Effects of partial hepatectomy on the plasma membrane status and the invertor mechanism of the hepatocyte Na,K-ATPase activity regulation in rats of various age. AB - The experiments were performed on adult (6-8 months) and old (22-24 months) Wistar rats. Insulin induced plasma membrane hyperpolarization and hepatocyte Na,K-ATPase activation in adult but not in old sham-operated rats. Partial hepatectomy had no effect on the invertor mechanism of Na,K-ATPase activity regulation in the liver of adult rats, while pronounced changes took place in old animals 4 weeks after partial hepatectomy. Insulin induced hyperpolarization in hepatocyte plasma membrane and activation of Na,K-ATPase both in old and adult hepatectomized rats. Invertors, intracellular regulators of the plasma membrane status, played an important role in the mechanism of this insulin-induced hyperpolarization. Four weeks after partial hepatectomy in old animals, the invertor mechanism of hepatocyte plasma membrane regulation appeared again, as well as membrane Na,K-ATPase capability to react to insulin action. PMID- 10386175 TI - Thalamic metabolic rate predicts EEG alpha power in healthy control subjects but not in depressed patients. AB - BACKGROUND: EEG alpha power has been demonstrated to be inversely related to mental activity and has subsequently been used as an indirect measure of brain activation. The hypothesis that the thalamus serves as a neuronal oscillator of alpha rhythms has been supported by studies in animals, but only minimally by studies in humans. METHODS: In the current study, PET-derived measures of regional glucose metabolism, EEG, and structural MRI were obtained from each participant to assess the relation between thalamic metabolic activity and alpha power in depressed patients and healthy controls. The thalamus was identified and drawn on each subject's MRI. The MRI was then co-registered to the corresponding PET scan and metabolic activity from the thalamus extracted. Thalamic activity was then correlated with a 30-min aggregated average of alpha EEG power. RESULTS: Robust inverse correlations were observed in the control data, indicating that greater thalamic metabolism is correlated with decreased alpha power. No relation was found in the depressed patient data. CONCLUSIONS: The results are discussed in the context of a possible abnormality in thalamocortical circuitry associated with depression. PMID- 10386176 TI - The efficacy of lamotrigine in rapid cycling and non-rapid cycling patients with bipolar disorder. AB - BACKGROUND: Patients with bipolar disorder (BD) who have rapid cycling features are often treatment refractory. Clear and conclusive evidence regarding effective treatments for this group is not available. METHODS: Patients with diagnoses of refractory bipolar disorder who were currently experiencing manic, mixed, depressive, or hypomanic episodes were treated with lamotrigine as add-on therapy (60 patients) or monotherapy (15 patients). We compared the efficacy of lamotrigine in the 41 rapid cycling and 34 non-rapid cycling patients with BD. RESULTS: Improvement from baseline to last visit was significant among both rapid cycling and non-rapid cycling patients for both depressive and manic symptomatology. For patients entering the study in a depressive episode, improvement in depressive symptomatology was equivalent in the two groups. Among patients entering the study in a manic, mixed, or hypomanic episode, those with rapid cycling improved less in manic symptomatology than did non-rapid cycling patients. Among rapid cycling patients with initial mild-to-moderate manic symptom severity, improvement was comparable to that in non-rapid cycling subjects; however, the subset of rapid cycling patients with severe initial manic symptomatology had little improvement in mania. Rapid cycling patients had earlier onset and more lifetime episodes of mania, depression, and mixed mania. CONCLUSIONS: Lamotrigine was generally effective and well tolerated in this group of previously non-responsive, rapid cycling bipolar patients. PMID- 10386177 TI - Donepezil in treatment-resistant bipolar disorder. AB - BACKGROUND: A considerable percentage of patients with bipolar disorder do not respond or do not tolerate conventional treatment. Cholinesterase (ChE) inhibitors have been suggested to possess depressogenic and antimanic properties. METHODS: We report a case series of treatment-resistant bipolar patients (n = 11) to whom we administered the ChE inhibitor donepezil. Four patients met criteria for current manic episode, 5 for mixed episode, 1 for hypomanic episode, and 1 for major depressive episode. Donepezil was added to current medication on an openlabel basis. Ratings were based on a retrospective chart review. RESULTS: Of the 11 patients, 6 (54.5%) demonstrated marked improvement (improvement in CGI-S > or = 2), 3 (27.2%) demonstrated slight improvement, 1 did not respond, and 1 did not tolerate the medication. Among those patients who had marked improvement (i.e., responders, n = 6), improvement was observed within 2 weeks or less in 5 of them (83%). Patients experienced only minor side effects. CONCLUSIONS: These pilot data suggest the efficacy and safety of donepezil in the treatment of bipolar disorder. To our knowledge this is the first published report on the use of donepezil in the treatment of mood disorders. Controlled, randomized, double blind studies are necessary to validate these preliminary observations. PMID- 10386178 TI - Hyperintense lesions on magnetic resonance images in bipolar disorder. AB - BACKGROUND: To examine the magnetic resonance (MR) images of bipolar patients across a wide age range for the presence of hyperintense lesions compared to age- and gender-matched control subjects. METHODS: Consecutive admissions to a mood disorders unit over a 2-year period were evaluated retrospectively for the presence of bipolar disorder by DSM-III-R criteria and whether they received an MR scan. Bipolar patients (n = 70, mean age = 49.9 +/- 19.7 years) were age- and gender-matched to control subjects (n = 70, mean age = 53.2 +/- 18.1 years) and the MR scans were rated to assess for the presence of hyperintensites. RESULTS: Compared to control subjects, the bipolar patients demonstrated hyperintense lesions in the subependymal region, subcortical gray nuclei, and the deep white matter. CONCLUSIONS: Hyperintense lesions in bipolar patients are found in both the subcortical white matter and gray nuclei and may play an important role in the etiology of bipolar illness. PMID- 10386179 TI - Effect of catecholamine depletion on lithium-induced long-term remission of bipolar disorder. AB - BACKGROUND: This study investigated the effects of catecholamine depletion with alpha-methylparatyrosine (AMPT) on mood indices in patients with bipolar disorder who were in long-term remission with lithium therapy. METHODS: Eight subjects with DSM-IV bipolar disorder currently in remission for > 3 months on lithium were included in the study. Subjects were given either AMPT or placebo, in a randomized double-blind manner, in two test sessions of 4 days each. RESULTS: Subjects did not have any significant changes in mood during AMPT or placebo administration; however, 24-48 hours after the last active AMPT dose subjects had a transient relapse of hypomanic symptoms. Relapse of hypomanic symptoms did not correlate with increases in serum levels of homovanillic acid or 3-methoxy-4 hydroxyphenylglycol. CONCLUSIONS: These findings suggest that the mechanism of prevention of manic relapse by long-term lithium therapy may be dependent on stability of the catecholamine system. PMID- 10386180 TI - Psychomotor performance and monoamine function in bipolar and unipolar affective disorders. AB - BACKGROUND: Affective disorders are associated with prominent psychomotor abnormalities that may be related to changes in arousal or motivation due to altered catecholamine function. METHODS: We investigated relationships between performance on psychomotor tests of motor speed (reaction time and tapping speed) and visual tracking (trail making and dot placement) and catecholamine system function including cerebrospinal fluid (CSF) or urinary concentrations of catecholamines or their metabolites. Subjects were medicine-free inpatients with unipolar depression or with manic, depressive, or mixed episodes of bipolar disorder, and healthy controls matched by gender and stratified by age. RESULTS: Unipolar and bipolar depressed patients were impaired in motor speed, dexterity, and visual tracking, whereas manic and mixed patients did not differ from controls. Tapping speed correlated positively with CSF 3-methoxy-4 hydroxyphenylglycol in healthy controls and with CSF homovanillic acid in bipolar depressed subjects. Increased catecholamine function correlated with slowing in all other measures for patients with bipolar disorder. Relationships between catecholamines and psychomotor function were weaker in unipolar depressed subjects. Psychomotor function was related to severity of depression in bipolar, but not in unipolar, patients. CONCLUSIONS: These data suggest that catecholamine systems are associated with increased arousal and psychomotor impairment in patients with bipolar disorder. Similar behavioral changes have different neurotransmitter relationships in unipolar disorder. PMID- 10386181 TI - Activation of indices of cell-mediated immunity in bipolar mania. AB - BACKGROUND: Evidence supports that macrophages as well as lymphocytes and their products may be involved in the pathophysiology of psychiatric disorders. Whether patients with bipolar disorder have activation or reduction of immunity during a manic episode remains unclear. METHODS: The purpose of this case-control study was to investigate the lymphocyte proliferation to phytohemagglutinin (PHA), concanavalin A, and pokeweed mitogen, and plasma levels of soluble interleukin-2 receptor (sIL-2R) and sIL-6R in patients with bipolar mania (DSM-III-R). The subjects were 23 physically healthy patients with Young Mania Rating Scale (YMRS) scores > or = 26 as well as aged < or = 45 years and 23 age- and gender-matched normal control subjects. The above immune variables were measured in acute mania and consequent remission (YMRS scores < or = 12) among bipolar patients. RESULTS: The lymphocyte proliferation to PHA and the plasma sIL-2R levels, but not sIL-6R, of bipolar patients were significantly higher in acute mania than in consequent remission. These elevations were not due to differences in medication status. Only in acute mania were the plasma sIL-2R levels of patients significantly higher than control subjects. A positive correlation between the changes of manic severity and plasma sIL-2R levels was observed. Remitted bipolar patients and normal control subjects did not differ in any of these measures. CONCLUSIONS: Cell-mediated immunity activation in bipolar mania was demonstrated and may be through a specifically state-dependent immune response. PMID- 10386182 TI - Lithium reduces tau phosphorylation: effects in living cells and in neurons at therapeutic concentrations. AB - BACKGROUND: The mechanism of action of lithium remains to be determined satisfactorily. Recent studies suggested a possible role in inhibiting glycogen synthase kinase-3 (GSK-3), previously shown to phosphorylate the protein tau. Tau is expressed mainly in neurons, where it functions to stabilize microtubules in a phosphorylation-dependent manner. METHODS: Neurons and transfected non-neuronal cells were treated with lithium and the phosphorylation of tau at multiple epitopes examined by western blotting and by immunocytochemistry. Using green fluorescent protein as a tag we examined the effects of lithium on phosphorylated tau in living cells. RESULTS: Lithium reversibly reduced tau phosphorylation at therapeutic concentrations, and even at high concentrations did not alter neuronal morphology. Green fluorescent protein tagged-tau when phosphorylated by GSK-3 was diffusely distributed; treatment with lithium resulted in association with microtubules and then bundle formation. Removing lithium allowed observation of the dissolution of bundles and gradual dissociation of tau from microtubules in living cells. CONCLUSIONS: Lithium may have multiple effects in brain, but at least one action is demonstrated to be a relative inhibition of GSK-3-induced tau phosphorylation. These results carry implications for future studies of the actions of mood-stabilizing drugs and indeed of the molecular mechanisms of affective disorders. PMID- 10386183 TI - Differential expression of thyroid hormone receptor isoforms by thyroid hormone and lithium in rat GH3 and B103 cells. AB - BACKGROUND: The interaction between lithium, a mood stabilizer, and the thyroid axis has been extensively studied; however, the regulation of thyroid hormone receptors by lithium is yet to be investigated. METHODS: To test whether lithium affects thyroid hormones at the receptor level, we examined the effects of lithium in combination with triiodothyronine (T3) on gene expression of thyroid hormone receptor isoforms in GH3 and B103 cells. RESULTS: The pattern of expression as well as the magnitude of regulation of the different thyroid hormone receptor isoforms appeared to be cell line specific. Whereas T3 regulated all four isoforms in GH3 cells at both time points, T3 did not alter thyroid hormone receptor TR alpha 1 and TR alpha 2 mRNA in B103 cells. Addition of lithium to thyroid hormone-deficient GH3 cells decreased TR alpha 1, alpha 2, and beta 2 expression without affecting TR beta 1 expression at 2 but not 5 days. Addition of lithium to T3-treated GH3 cells did not further modulate gene expression of TR alpha 1, alpha 2, beta 1, or beta 2 when compared to cells treated with T3 alone. The effects of lithium in B103 cells appeared to be isoform specific as well as time dependent, since TR alpha 1 expression was selectively decreased in B103 cells, when treated with T3 in the presence of lithium. CONCLUSIONS: The present study provides direct evidence that T3 and/or lithium regulate TR gene expression in vitro in a both time-dependent and cell line-specific manner. PMID- 10386184 TI - The prevalence of seasonal affective disorder in The Netherlands: a prospective and retrospective study of seasonal mood variation in the general population. AB - BACKGROUND: The aim of the present study was to assess the prevalence of seasonal affective disorder (SAD) in The Netherlands. METHODS: The subjects (n = 5356), randomly selected from community registers, were given the Seasonal Pattern Assessment Questionnaire and the Centre for Epidemiological Studies Depression Scale over a period of 13 months. The response rate was 52.6%. RESULTS: Three percent of the respondents met the criteria for winter SAD, 0.1% for summer SAD. The criteria for subsyndromal SAD, a milder form of SAD, were met by 8.5%, 0.3% of whom showed a summer pattern. Younger women received a diagnosis of SAD more often than men or older women. CONCLUSIONS: SAD subjects were significantly more often unemployed or on sick leave than other subjects. Respondents who met winter SAD criteria were significantly more depressed than healthy subjects, in both winter and summer. Finally, month of completion had no influence on the number of subjects meeting the SAD criteria. PMID- 10386185 TI - Patterns of DST positivity in remitted affective disorders. AB - BACKGROUND: While the Dexamethasone Suppression Test (DST) has been extensively used in cross-sectional observations of patients with major affective disorders, studies have tended to ignore the longitudinal application of the DST in patients stabilized on long-term prophylactic medication. METHODS: Monthly DST's were performed on 19 patients, 16 with bipolar disorder and 3 with recurrent major depression. All cases had an excellent response to lithium treatment, and family history positive for bipolar disorder. The average duration of observation was 4 years. RESULTS: All patients remained clinically stable throughout the period of observation. Eleven patients showed intermittent DST positivity ranging from 10% to 60% of tests, and 2 patients exhibited no positivity. Six patients had fewer than 10% positive DST's. Females showed significantly higher positivity than males. The frequency of positivity did not correlate with current age, age of illness onset, duration of illness, duration of lithium treatment, or season. The risk of primary affective disorders in first-degree relatives was also unrelated to the frequency of positivity. CONCLUSIONS: While the highly selected and small sample population limits generalizability, our observations suggest that clinically sufficient lithium prophylaxis does not automatically prevent intermittent HPA dysregulation. We hope that a better understanding of this phenomenon will offer new approaches to the long-term management of mood disorders. PMID- 10386186 TI - Evidence of a possible role of altered angiotensin function in the treatment, but not etiology, of depression. AB - BACKGROUND: Angiotensin-converting enzyme inhibitors are reportedly effective in the treatment of depression; furthermore, antidepressants decrease angiotensin function. It appears therefore that reduced angiotensin function may be important in the treatment of depression. The aims of this study were to elucidate the actions of antidepressants on angiotensin receptors; to investigate the antidepressant potential of an angiotensin antagonist; and to study angiotensin receptors in depressed puerperal women. METHODS: The effects of antidepressant drugs on angiotensin receptors and the relationship between mood and platelet receptors in puerperal women were investigated using radioligand binding. The antidepressant potential of the angiotensin antagonist losartan was assessed using the mouse forced swim test. RESULTS: Desipramine, but neither fluoxetine nor tranylcypromine, displaced angiotensin from its receptor; however, there was no significant relationship between receptor number and depressed mood. In the forced swim test losartan was shown to possess antidepressant like activity. CONCLUSIONS: These findings indicate that antidepressants differ in the mechanism by which they reduce angiotensin function, but the link between antidepressants and angiotensin is reiterated by the demonstration that losartan possesses antidepressant like activity. There is, however, no evidence of abnormal angiotensin receptors in women with depressed mood postpartum. PMID- 10386187 TI - Platelet cytosolic calcium hyperresponsivity to serotonin in patients with hypertension and depressive symptoms. AB - BACKGROUND: Data from recent studies indicate that the presence of depression is an independent risk factor for cardiovascular and cerebrovascular events. The mechanism by which depression increases the morbidity and mortality risks in patients with comorbid vascular disease is currently the object of considerable research interest. Platelets may be involved in this pathological process. Although many investigators have extensively evaluated platelet biochemistry in depressed patients, there currently exists very little information regarding how the biochemical alterations might relate to an increased risk of cardiovascular events. In this study, we examined the responsivity of platelet cytosolic calcium concentrations ([Ca++]i) to serotonin stimulation in populations of hypertensive patients with or without comorbid depressive symptoms. METHODS: We utilized Fura 2 loaded platelets to compare changes in intracellular calcium levels (delta [Ca++]i) following serotonin stimulation among 48 patients with hypertension and varying degrees of depressive symptomatology. RESULTS: We found that those patients with higher scores on standardized depression rating scales showed significantly greater [Ca++]i (82.82 +/- 15.88 mmol/L) increase compared with [Ca++]i (60.10 +/- 22.65 mmol/L) patients with lower depression scores. CONCLUSIONS: The results of this study support the hypothesis that the enhanced platelet reactivity seen in patients with depressive symptoms may mediate the deleterious effects of depression on cardiovascular disease. PMID- 10386188 TI - Serotonin-induced platelet intracellular Ca2+ responses in untreated depressed patients and imipramine responders in remission. AB - BACKGROUND: Intracellular Ca2+ metabolism in platelets has been investigated as a peripheral marker of affective disorders. METHODS: We investigated the intracellular free Ca2+ concentration in platelets in both untreated depressed patients with no medications and patients in remission who were treated by imipramine (IMI) (IMI responders) using a Ca(2+)-sensitive fluorescent probe fura 2. RESULTS: The increases in intracellular free Ca2+ concentration in platelets induced by stimulation with serotonin (5-HT) ([Ca2+] delta) were significantly higher in both the untreated patients and the IMI responders compared with healthy controls; however, there were no significant differences in the basal Ca2+ levels in the platelets ([Ca2+]B) among the three groups. On the other hand, in the IMI responders, we observed positive correlations between the duration of the remission and [Ca2+]B, but not [Ca2+] delta. CONCLUSIONS: Our present data suggest that the enhancement of 5-HT2A-induced Ca2+ responses persisted after remission in depressed patients, and that the duration of the remission is a factor varying the intracellular basal Ca2+ levels. PMID- 10386189 TI - Lack of correlation between cerebrospinal fluid thyrotropin-releasing hormone (TRH) and TRH-stimulated thyroid-stimulating hormone in patients with depression. AB - BACKGROUND: It has been proposed that elevated central thyrotropin-releasing hormone (TRH) is associated with the blunted thyroid-stimulating hormone (TSH) response to TRH in patients with depression. Few studies have directly evaluated this relationship between central nervous system and peripheral endocrine systems in the same patient population. METHODS: 15 depressed patients (4 male, 11 female, 12 bipolar, and 3 unipolar) during a double-blind, medication-free period of at least 2 weeks duration, underwent a baseline lumbar puncture followed by a TRH stimulation test. Cerebrospinal fluid (CSF) TRH and serial serum TSH, free thyroxine, triiodothyronine, prolactin, and cortisol were measured. A blunted response to TRH was defined as a delta TSH less than 7 microU/mL. RESULTS: There was no significant difference in mean CSF TRH between "blunters" (2.82 +/- 1.36 pg/mL) and "non-blunters" (3.97 +/- 0.62 pg/mL, p = .40). There was no evidence of an inverse relationship between CSF TRH and baseline or delta TSH. There was no correlation between CSF TRH and the severity of depression or any other endocrine measure. CONCLUSIONS: These data are not consistent with the prediction of hypothalamic TRH hypersecretion and subsequent pituitary down-regulation in depression; however, CSF TRH may be from a nonparaventricular nucleus hypothalamic source (i.e., limbic area, suprachiasmatic nucleus, brain stem dorsal raphe) and thus, not necessarily related to peripheral neuroendocrine indices. PMID- 10386190 TI - Peripheral thyroid hormone levels in treatment resistant depression. AB - BACKGROUND: Various abnormalities of thyroid function have been inconsistently reported in major depression. The inconsistency between studies may be due to several factors including the stage of treatment resistance. METHODS: One hundred and one patients with major depressive disorder receiving their first antidepressant for their current major depressant episode had baseline thyroid function test performed. On completion of treatment, their stage of antidepressant resistance was determined. RESULTS: Severity of depression but not any peripheral thyroid hormone level was associated with stage of anti-depressant treatment resistance. CONCLUSIONS: Stage of treatment resistance does not appear to be a factor in the variability in peripheral thyroid hormone levels in unipolar major depression. PMID- 10386191 TI - First-cycle REM density in never-depressed subjects with borderline personality disorder. AB - BACKGROUND: There is much interest in the identification of polysomnographic markers of liability to the mood disorders that may predate the onset of illness in high-risk subjects, and/or remain altered after remission. One such putative marker is rapid eye movement (REM) density during the first REM period. METHODS: Never-depressed subjects with borderline personality disorder (BPD) as a group at high risk for the mood disorders were compared by continuous 48-hour ambulatory electroencephalographic monitoring to age- and gender-matched controls. RESULTS: Subjects with BPD had significantly higher REM density during the first REM period. One man with BPD who later committed suicide had REM density values exceeding the mean value of his group by 2 SD. CONCLUSIONS: These data extend the view that REM density in the first REM period can be a marker of liability to the mood disorders, as it is present in a group of young subjects at heightened risk for depression. PMID- 10386192 TI - A study of platelet serotonin receptor in mania. AB - BACKGROUND: Serotonergic (5-HT) dysfunction has been hypothesized in mania; however platelet studies on the 5-HT uptake rate and the 5-HT transporter have revealed inconsistent results. To the best of our knowledge no studies have been conducted on the 5-HT2 receptor status in mania. METHODS: We determined density (Bmax) and dissociation constant (Kd) of 5-HT2 receptors in the platelets of 29 normal control and 29 manic subjects using 125I-ketanserin as the binding radioligand. The manic patients were assessed for the same after 14 days of treatment with lithium (n = 14) and after return to premorbid levels of functioning (n = 5). RESULTS: There were no significant differences in the Bmax (3.51 +/- 3.04 vs. 3.14 +/- 3.44 fmol/mg protein, p = ms) values between normal control and manic subjects. In comparison to the baseline values Bmax at day 14 (3.49 +/- 3.68 vs. 2.18 +/- 1.90 fmol/mg protein, p = ns) and following recovery (1.17 +/- 0.85 vs. 1.29 +/- 1.13 fmol/mg protein, p = ns) did not show any significant difference. CONCLUSIONS: Our findings preliminarily suggest that the platelet 5-HT2 receptor is neither a state marker nor a trait marker in mania; however studies on the 5-HT2 receptor using positron-emission tomography ligands will help in conclusively ruling out the involvement of this receptor in mania. PMID- 10386193 TI - Brain computed tomography in geriatric manic disorder. AB - BACKGROUND: Excess brain changes in geriatric manic patients have been hypothesized. Few neuroimaging studies are available. METHODS: Brain computed tomography scans in geriatric patients with manic disorder (n = 30) were compared to those in same-age control subjects (n = 18). Ratings of cortical sulcal widening (CSW), lateral ventricle-brain ratio (VBR), and related linear measures were determined. RESULTS: Patients had greater CSW scores (Exact p = .002) and VBR (t = 2.51, df = 46, p < .02) compared to control subjects. CSW was positively associated with age at illness onset (rs = .46, p < .01) and age at first manic episode (rs = .53, p < .005). VBR was poorly correlated with CSW and was not associated with these indices of illness course. CONCLUSIONS: These findings support the need for further investigation of relationships between brain structure and clinical features in geriatric mania. PMID- 10386194 TI - Low blood cholesterol and low platelet serotonin levels in violent suicide attempters. AB - AIM OF THE STUDY: This study investigated the possible connection between serum cholesterol levels and platelet serotonin (5-HT) content in violent suicide attempters and matched controls. METHODS: Blood samples for cholesterol and platelet 5-HT levels were obtained from 17 drug-free patients within 3 days after the suicide attempt. RESULTS: Serum cholesterol and platelet 5-HT levels in the suicide attempters were significantly lower than in the controls; however, we did not find any significant correlation between these two variables. Indeed, three clinical dimensions are present in this patient group: suicidality, violence, and impulsiveness. Because we did not find a difference in cholesterol and platelet 5 HT levels between impulsive and nonimpulsive patients, these two indexes may more reflect the dimension of suicidality and/or violence. CONCLUSIONS: Further investigation is necessary to study the dependence of these two peripheral abnormalities within the context of violent suicidal behavior. PMID- 10386195 TI - Antiglucocorticoid treatment of depression: double-blind ketoconazole. AB - BACKGROUND: Hypercortisolemia is frequently observed in major depression but its pathophysiologic significance is unknown. In patients in whom hypercortisolism contributes to depressive symptomatology, antiglucocorticoid agents should have antidepressant effects. METHODS: Twenty medication-free depressed patients (eight of whom were hypercortisolemic and twelve of whom were not) received either the cortisol biosynthesis inhibitor, ketoconazole (400-800 mg/d p.o.) or placebo for 4 weeks in a double-blind manner, and behavioral ratings were performed weekly. RESULTS: Ketoconazole, compared to placebo, was associated with improvements in depression ratings in the hypercortisolemic, but not in the non-hypercortisolemic patients. The hormonal changes seen (decreased dehydroepiandrosterone and testosterone levels and increased pregnenolone and pregnenolone-sulfate levels) are consistent with enzymatic blockade of C17,20-lyase, 11-hydroxylase, and 17 hydroxylase. Ketoconazole was generally well tolerated with no occurrence of significant side effects or laboratory abnormalities. CONCLUSIONS: This small scale double-blind study suggests that antiglucocorticoids have antidepressant activity in hypercortisolemic depressed patients. The data are consistent with a causal role of adrenocortical dysfunction in some depressed patients and suggest the need for larger-scale trials. PMID- 10386196 TI - A rapid-cycling bipolar patient treated with long nights, bedrest, and light. AB - BACKGROUND: Stabilization of rapid-cycling bipolar disorder is extremely difficult. METHODS: A refractory bipolar I rapid-cycling patient on valproate was treated with long "nights" (extended sleep in darkness) and daytime light therapy. RESULTS: Rapid cycling immediately stopped on initiation of a 10 hour dark/rest period. This was extended to 14 hours (plus a self-selected 1 hour midday nap) without problems. Depression gradually improved when midday light therapy was added; near-euthymia was attained after light therapy was shifted to the morning. CONCLUSIONS: Nonpharmacological chronobiological treatments may be a means to interrupt rapid cycling. PMID- 10386197 TI - Treatment response to prophylactic lithium and family history of psychiatric disorders. PMID- 10386198 TI - Family history of affective disorders and the significance for prophylactic effect of lithium treatment. PMID- 10386199 TI - Occupational asthma. PMID- 10386200 TI - Documenting informed consent for antipsychotic medication. PMID- 10386201 TI - Need for an Institute of Primary Care Research within the Canadian Institutes of Health Research. PMID- 10386202 TI - Some provincial chapters adopting a more political stance. PMID- 10386203 TI - Unsympathetic to physician shortage. PMID- 10386204 TI - Teaching activities for aboriginal health issues. PMID- 10386205 TI - Safe use of valproic acid during pregnancy. AB - QUESTION: I have an epileptic patient who plans pregnancy. It took us years to control her seizures, and valproic acid seems to be the only way to control them. What is the risk to her fetus? ANSWER: Neural tube defects (NTDs) are the most common of the major anomalies associated with in utero exposure to valproic acid. About 1% to 2% of exposed fetuses suffer adverse effects. PMID- 10386206 TI - Ophthaproblem. Purtscher's disease. PMID- 10386207 TI - Dermacase. Crusted Norwegian scabies. PMID- 10386208 TI - Increased risk of mortality in people older than 65. PMID- 10386209 TI - Radiology rounds. Bilateral spondylolysis at L5 with associated spondylolisthesis. PMID- 10386210 TI - Practice tips. Simple method for curing hiccups. PMID- 10386211 TI - Outcome of pregnancy following mothers' use of new SSRIs. PMID- 10386212 TI - Is carotid endarterectomy beneficial for patients with stenosis? PMID- 10386213 TI - Zolmitriptan. AB - *Zolmitriptan (Zomig) is an antimigraine drug similar to sumatriptan.*The clinical file mainly comprises placebo-controlled, dose-finding studies recommending an optimal oral dose of 2.5 mg.*Zolmitriptan has been compared with sumatriptan in a trial that showed no difference in efficacy. In particular, the recurrence rate of headache after initial relief was not lower on zolmitriptan than on sumatriptan.*The safety profile of zolmitriptan is similar to that of sumatriptan. The contraindications relating to a history of cardiovascular disease must be respected because of the vasoconstrictive effect of the drug.*Zolmitriptan has the same drug interactions as sumatriptan. Zolmitriptan should not be used during migraine attacks by patients using propranolol. PMID- 10386214 TI - Informed consent for antipsychotic medication. AB - OBJECTIVE: To determine family physicians' attitudes and practices regarding documentation of informed consent for antipsychotic medication. DESIGN: Pilot cross-sectional study. SETTING: Teaching and non-teaching hospitals in Toronto, Ont. PARTICIPANTS: Thirty family physicians were selected in equal numbers from teaching and non-teaching hospitals with no more than five physicians from a given hospital. Participants were treating at least 10 patients with antipsychotic medication. Participants' mean age was 44.3 years; 83% were men. MAIN OUTCOME MEASURES: Documentation of consent and of disclosure of consent for antipsychotic medication in patients' charts. RESULTS: Documentation was found in only 13% of charts. Whether it was there or not did not correlate with information disclosed, score on an attitude scale, or demographics. Physicians who found documentation time-consuming were less likely to document. Most physicians disclosed reasons for antipsychotic medication, but less than half described tardive dyskinesia, a potentially irreversible movement disorder that affects about 25% of patients on long-term treatment. CONCLUSIONS: The low rate of documentation observed in this sample was consistent with reports of similar samples and might indicate that family physicians are unaware of recommendations for documentation or simply do not have time to keep abreast of current recommendations. Many physicians thought signed consent forms unnecessary for psychotic patients, and even more believed seeking consent for antipsychotic medications would increase patient anxiety. PMID- 10386215 TI - Peer consultation reflection exercise. AB - OBJECTIVE: To explore participants' overall perception of the value of the Peer Consultation Reflection Exercise (PCRE); of barriers and facilitators to participation and learning during a PCRE; and of the transferability of the experience to participants' own settings. DESIGN: This study used the qualitative techniques of key informant interviews and a focus group. SETTING: Focus group and key informant interviews at the 1996 Annual Meeting of the College of Family Physicians of Canada's Section of Teachers. PARTICIPANTS: Family medicine teachers attending a PCRE. METHOD: Five key informant interviews and one focus group composed of five participants were conducted to explore participants' experience of participating and learning during a PCRE. MAIN FINDINGS: Participants viewed the PCRE as a valuable opportunity to interact and learn from colleagues a were especially impressed with the opportunity to listen. Confidentiality and the important role of the facilitator were identified as key components. The greatest perceived barrier was the formal structure of the PCRE. CONCLUSIONS: The PCRE is an innovative strategy for personal and professional development. It could be used in other settings. PMID- 10386216 TI - Occupational allergies and asthma. AB - OBJECTIVE: To review aspects of occupational allergies and asthma for primary care physicians recognizing, diagnosing, and managing patients with these conditions. QUALITY OF EVIDENCE: Studies in the medical literature mainly provide level 2 evidence, that is, from at least one well-designed clinical trial without randomization, from cohort or case-control analytical studies, from multiple time series, or from dramatic results in uncontrolled experiments. MAIN MESSAGE: Occupational allergies and asthma have the best prognosis with an early, accurate diagnosis and subsequent avoidance of exposure to the relevant sensitizer. These diagnoses can normally be suspected from the clinical history. Primary care physicians can also initiate investigations to make an objective diagnosis, can assess workplace exposure agents from the history, and can review appropriate data sheets on material safety. Specialist evaluation is likely to be needed for skin tests, however, and for other specialized tests (such as pulmonary function assessments at work and off work or specific challenges with the suspected workplace agent). Patients with a confirmed diagnosis need appropriate medical management of their allergic manifestations or asthma, but also often require psychosocial support during the period of investigation and management, especially in relation to required changes in their work and to compensation or insurance claims. CONCLUSIONS: Consider workplace exposure as a source of patients' allergies or asthma and aim to make an early, accurate diagnosis. PMID- 10386217 TI - Case report: pneumothorax and asthma. PMID- 10386218 TI - Managing elderly people's osteoporosis. Why? Who? How? AB - OBJECTIVE: To guide family physicians through assessment of why treating elderly people's osteoporosis is necessary, who to treat, and how to treat in a practical way. QUALITY OF EVIDENCE: Evidence of the efficacy of treatment for osteoporosis is shown by a reduced probability of fracture. This can be ascertained by direct evaluation for bisphosphonates, calcium, and calcitonin, or indirectly by ascertaining benefit to bone mineral density for hormone replacement therapy (HRT) and exercise. MAIN MESSAGE: Unless medically contraindicated, all elderly people should take supplementary vitamin D (800 IU/d) and calcium (1500 mg/d). Those with risk factors for osteoporosis (e.g., smoking, thinness, previous fracture when older than 50 years, fracture in first-degree relatives older than 50 years, and steroid use) should have a bone density measurement. Those meeting World Health Organization criteria for osteoporosis should also be treated with HRT or bisphosphonates or possibly with selective estrogen receptor modulators. CONCLUSIONS: Good evidence indicates that adequate treatment of osteoporosis can prevent fractures and thus reduce associated morbidity and mortality among vulnerable elderly people. Because of the prevalence of osteoporosis, the onus falls on family physicians to be the front-line managers. PMID- 10386219 TI - Highlights of the Canadian Thoracic Society guidelines for occupational asthma. PMID- 10386220 TI - Phaseout of chlorofluorocarbons (CFCs) in metered-dose inhalers. Highlights of the Canadian initial transition strategy. PMID- 10386221 TI - Millennium malady. Part 1: Commentary. PMID- 10386222 TI - Millennium malady. Part 2: Diagnostic and treatment plan. PMID- 10386223 TI - Urologist warns against automatic prescriptions. PMID- 10386224 TI - Good sex is more than just a good erection. PMID- 10386225 TI - Biological and clinical roles of adrenomedullin in circulation control and cardiovascular diseases. AB - 1. Adrenomedullin (AM) is found ubiquitously in tissues and organs, especially in cardiovascular tissues and in the kidney, lung and endocrine glands. It has multifunctional biological properties, of which, its effects on the control of circulation and body fluid volume regulation seem to be the most outstanding and characteristic. 2. Acute administration of a high dose of AM induces a vasodilator depressor response, cardiac inotropic effects, diuresis and suppression of aldosterone secretion in experimental animals. 3. Long-term continuous administration of a very low dose of AM causes vasodilation in sheep (0.5 microgram/kg per h) and hypotension in rats (0.8 microgram/kg per h). 4. The plasma concentration of AM increases under pathological conditions such as congestive heart failure, myocardial infarction and hypertensive and renal diseases. Under these disease conditions, AM may be produced in vascular endothelial and smooth muscle cells and in cardiac myocytes in response to volume expansion, hypertension and activated humoral factors, such as catecholamine and the renin-angiotensin system. 5. Increased AM in the circulating blood and cardiovascular tissues may counteract pathological deviation in the system that controls circulation and body fluid volume, acting against cardiovascular damage and disease. 6. Because of these beneficial properties in the cardiovascular system, AM and its pharmaceutical ligands should prove useful in the treatment of cardiovascular diseases. PMID- 10386226 TI - Kinetic aspects of drug disposition in the lungs. AB - 1. The pharmacokinetic role of the lungs has been extensively studied using in vitro preparations, but this information has not been well integrated into many systemic pharmacokinetic models. 2. The lung is characterized by short diffusion distances, extremely high relative perfusion and heterogeneous cell types. Anionic and neutral lipophilic drugs have relatively small distribution volumes in the lungs due to their low lipid content. Cationic lipophilic drugs can accumulate in the lungs, probably due to trapping in mitochondria and lysosomes, forming very slowly eluting pools. 3. Drug metabolism in the lungs is possible, but not universal. The lung, generally, has a low activity for many of the metabolic enzymes found in the liver, although this activity is relatively more inducible. The resultant drug extraction would be 'enzyme limited', variable and flow dependent. 4. Double indicator studies of first-pass lung kinetics can characterize short-term distribution in the lungs, but not longer-term distribution or metabolism; the converse applies for studies of drug concentration gradients across the lungs. No single study or model has adequately defined the short- and long-term kinetics of drugs in the lungs. 5. Drug clearance in the lungs can contribute to an apparent total body clearance in excess of hepatic blood flow and cardiac output. The lung is a first pass filter for any drug administered on the venous side of the circulation and can act as a 'capacitor' that damps the first-pass concentration peak in the blood after intravenous bolus injection. PMID- 10386227 TI - Actions of endothelin-1 on cultured rat renomedullary interstitial cells are modulated by nitric oxide. AB - 1. Cultured renomedullary interstitial cells (RMIC) isolated from 4-week-old Sprague-Dawley rat kidneys possess ETA receptors, as identified by reverse transcription-polymerase chain reaction (RT-PCR). 2. Treatment with endothelin (ET)-1 (10(-6) mol/L) increases the intracellular inositol 1,4,5-trisphosphate concentrations within 10 s and intracellular calcium concentrations after 7 s. 3. Endothelin-1 (10(-7) and 10(-10) mol/L) induced increases in intracellular cAMP concentrations, but only in the presence of N omega-nitro-L-arginine, a nitric oxide synthase (NOS) inhibitor. Addition of ET-1 (10(-10) mol/L) to the RMIC culture led to increases in intracellular cGMP concentrations through activation of NOS. 4. In the presence of ET-1 (10(-7) and 10(-10) mol/L) and during NOS inhibition, RMIC responded with increased cell proliferation and extracellular matrix (ECM) synthesis. These responses were abolished by BQ-123 (10(-6) mol/L), suggesting mediation via the ETA receptor subtype. The proliferative effect of ET 1 was also abolished by atrial natriuretic peptide (10(-6) mol/L). 5. The present study provides evidence that binding of ET-1 to ETA receptors on RMIC activates several intracellular second messenger systems that mediate cell proliferation and ECM synthesis. 6. These results also highlight an important interaction between ET-1 and nitric oxide in the control of RMIC function. PMID- 10386228 TI - Pharmacological profile of orally administered YM087, a vasopressin antagonist, in conscious rats. AB - 1. YM087 is a newly synthesized non-peptide arginine vasopressin (AVP) antagonist that shows high affinity for both V1A and V2 receptors. In the present study, the V1A and V2 receptor antagonist effects of orally administered YM087 were assessed in conscious rats. 2. In conscious rats, orally administered YM087 (0.1, 0.3 and 1.0 mg/kg) did not affect basal blood pressure, but YM087 dose-dependently inhibited 30 mU/kg, i.v., AVP-induced pressor responses. This inhibition lasted for over 8 h following the oral administration of the highest dose of YM087 (1 mg/kg). 3. In rats deprived of water and food for 16-18 h, oral administration of YM087 (0.1, 0.3, 1 and 3 mg/kg) dose-dependently increased urine volume and reduced urine osmolality, with associated increases in urinary sodium and potassium excretion. However, these increases in electrolyte excretion were lower than those seen at comparable diuretic doses of furosemide (3, 10, 30 and 100 mg/kg, p.o.). 4. Oral administration of YM087 (0.3, 1 and 3 mg/kg) produced a dose-dependent increase in urine volume in rats allowed free access to water, with the diuretic effect peaking 2-4 h post-dosing at all dose levels. The diuretic effect of YM087 was sustained 8-10 h after a dose of 3 mg/kg; this is in contrast with the transient diuresis seen after furosemide (100 mg/kg, p.o.) dosing. 5. The present results demonstrate that YM087 is an orally active AVP antagonist with potent and long-lasting effects. YM087 suppressed V1A receptor mediated pressor responses to AVP with minimal effects on basal haemodynamics and exerted a diuretic effect without increased electrolyte excretion by inhibiting V2 receptor-mediated water reabsorption. PMID- 10386229 TI - Sublingual nifedipine in elderly patients: even a low dose induces myocardial ischaemia. AB - 1. Low doses of sublingual nifedipine are still used for the treatment of hypertensive crises, although recent studies have raised concerns that sublingual nifedipine may cause serious dose-dependent adverse effects. The present study was performed to test the safety of low-dose sublingual nifedipine administered to elderly hypertensive patients. 2. Systemic blood pressure measurements and electrocardiographic (ECG) examinations were performed before and 45-60 min after a 5 mg dose of sublingual nifedipine in 93 consecutive hypertensive patients, 65 years of age or older, who were without coronary artery disease. In 33 patients, the effects of nifedipine on myocardial lactate metabolism were studied during cardiac catheterization. 3. In all patients, following nifedipine administration, blood pressure (BP) decreased significantly, while heart rate (HR) increased, and symptoms associated with elevated BP disappeared. However, changes consistent with myocardial ischaemia appeared on the ECG in six of 55 patients with left ventricular hypertrophy (LVH) and in one of 38 patients without LVH, although only two of these seven patients experienced angina-like precordial tightness. Sublingual nifedipine decreased myocardial lactate extraction from 52 +/- 13 to 38 +/- 19% in 20 patients with LVH (P = 0.02), but myocardial lactate extraction remained stable in 13 patients without LVH (49 +/- 7 to 50 +/- 5%; NS). The change in lactate extraction was significantly correlated with the percentage change in diastolic arterial pressure (r = 0.77, P < 0.001). 4. These results suggest that sublingual nifedipine, even at the low dose of 5 mg, may cause myocardial ischaemia in some elderly patients with LVH that is associated with a marked reduction in coronary perfusion pressure. PMID- 10386230 TI - Effects of heparin on vascular dysfunction in diabetic rats. AB - 1. Treatment with heparin has beneficial effects in diabetic nephropathy. The occurrence of increased urinary albumin excretion in diabetic patients reflects general vascular dysfunction, including increased transcapillary permeability of macromolecules. The aim of the present study was to evaluate the effects of heparin on vascular dysfunction in diabetic rats. 2. Male Sprague-Dawley rats were used in two studies. Diabetes was induced by 65 mg/kg, i.v., streptozotocin. In one study, diabetic rats were dosed subcutaneously with different heparin fractions for 8 months and the transcapillary escape rate of albumin (TERalb) was measured in anaesthetized animals. In the other study, heparin was given for 6 weeks, followed by tissue albumin clearance measurements in awake rats. Normal and diabetic rats receiving saline served as controls. 3. Blood glucose did not differ among the diabetic groups and ranged from 22 to 26 mmol/L. The mean (+/- SD) TERalb was increased by diabetes compared with values in normal rats (17.5 +/ 3 vs 14.1 +/- 3.3%/h, respectively). Neither unfractionated nor low molecular weight heparin significantly affected this increase. [131I]-Albumin clearance was significantly increased in diabetic rats in the eye, skin and skeletal muscle tissues compared with normal rats (0.17-0.40 vs 0.1-0.23 microL plasma/g per min). Low molecular weight heparin treatment did not affect the increased organ albumin clearance. 4. In conclusion, heparin treatment does not affect diabetes induced vascular dysfunction as expressed by increased TERalb and clearance of albumin in rats. PMID- 10386231 TI - Effects of mild aerobic exercise and a mild hypocaloric diet on plasma leptin in sedentary women. AB - 1. The present study was conducted to investigate whether mild aerobic exercise and a mild hypocaloric diet, instead of severe restrictions on caloric intake, would affect weight reduction and plasma leptin concentrations. 2. Forty-one middle-aged sedentary women (15 obese and 26 non-obese) participated in a 12 week lifestyle-modification programme to reduce cardiovascular risk factors. Bodyweight, body composition, plasma leptin concentrations, serum lipid profiles, fasting plasma glucose and fasting plasma insulin were measured before and after the 12 week intervention. The intervention consisted of aerobic exercise, corresponding to approximately 50% of maximal oxygen consumption, and personal diet counselling. 3. Bodyweight decreased by (mean +/- SD) 3.9 +/- 3.4 kg in the obese group (P < 0.05) and by 1.7 +/- 1.8 kg in the non-obese group (P < 0.05). The plasma leptin concentration decreased significantly from 14.7 +/- 5.3 to 8.9 +/- 3.6 ng/mL in the obese group (P < 0.001) and from 7.6 +/- 3.9 to 5.6 +/- 2.2 ng/mL in the non-obese group (P < 0.01). 4. Overall, for all subjects, both pre- and postintervention, the plasma leptin concentration was significantly correlated with body mass index (BMI; pre-intervention: r = 0.73, P < 0.0001; postintervention: r = 0.67, P < 0.0001), fat mass (FM; pre-intervention: r = 0.74, P < 0.0001; postintervention: r = 0.63, P < 0.0001) and fasting plasma insulin (pre-intervention: r = 0.66, P < 0.001; postintervention: r = 0.45, P < 0.01). The change in plasma leptin concentration was significantly correlated with the respective changes in BMI (r = 0.64, P < 0.0001), FM (r = 0.48, P < 0.01) and fasting plasma insulin (r = 0.58, P < 0.0001). Interestingly, the ratio of plasma leptin concentration to BMI or FM diminished significantly after intervention. In addition, we found that the plasma leptin concentration decreased in participants whose FM did not decrease. These results suggest that the production of leptin per unit FM decreased after intervention. 5. Mild aerobic exercise and a mild hypocaloric intake decreased body mass and the plasma leptin level in Japanese middle-aged sedentary women. This decrease in plasma leptin levels was likely to be associated with weight reduction plus some unknown factor(s). PMID- 10386232 TI - Effects of anti-Parkinsonian drugs on neurobehavioural changes induced by bilateral 6-hydroxydopamine lesions in rats. AB - 1. Effects of anti-Parkinsonian drugs on neurobehavioural changes induced by bilateral lesions of dopaminergic neurons were investigated in rats. 2. Dopaminergic neurons in rats were lesioned bilaterally by injection of 6 hydroxydopamine (6-OHDA; 8 micrograms) into the medial forebrain bundle at the level of the posterolateral hypothalamus. As a result, a decrease in locomotor activity and marked catalepsy and prolongation of grasping time were observed. 3. Levodopa, talipexole, bromocriptine and theophylline dose-dependently antagonized the decrease in locomotor activity induced by bilateral 6-OHDA lesions. These drugs also showed antagonistic effects on the appearance of catalepsy and prolongation of grasping time induced by bilateral 6-OHDA lesions. In contrast, trihexyphenidyl showed no antagonizing effect on the neurobehavioural changes induced by 6-OHDA lesions at any concentration tested. 4. Combined treatment with levodopa and talipexole antagonized the neurobehavioural changes induced by bilateral 6-OHDA lesions, whereas no marked changes were observed when either drug was administered separately. The same findings were noted with the simultaneous use of either levodopa (2 mg/kg) and theophylline (2 mg/kg) or talipexole (0.005 mg/kg) and theophylline (2 mg/kg). 5. These results indicate that this model may be useful for estimating the effects of drugs in the treatment of Parkinson's disease. PMID- 10386233 TI - Age-related changes in platelet microviscosity and aggregation in rats. AB - 1. To study possible changes in platelet microviscosity in aged animals, 1,6 diphenyl-1,3,5-hexatriene (DPH) was used as a nonpolar probe embedded in thrice washed platelets from young, adult and aged rats. With the known values of maximum limiting anisotropy and the structural parameter of DPH and by estimating the steady state of fluorescence anisotropy and the average fluorescence of lifetime, we applied the Perrin equation to calculate the microviscosity. 2. We measured platelet aggregation, platelet lipids and platelet polyunsaturated fatty acids to determine any causal relationship between these parameters. Platelet aggregation, the platelet molar ratio of cholesterol to phospholipid ([C]/[PL]) and platelet microviscosity increased with age (P < 0.05) and were correlated with one another (P < 0.05). 3. Age-dependent increases in the steady state of fluorescence anisotropy, order parameters and the short component of fluorescence lifetime of the fluorophore were expressed as functions of variables, such as microviscosity or the [C]/[PL] ratio. 4. Platelet concentrations of arachidonic and eicosapentaenoic acids increased with age, but were not associated with aggregation. Age-related changes in microviscosity and the [C]/[PL] ratio seemed to be determinants affecting biophysical properties of platelet aggregation. PMID- 10386234 TI - Effects of naloxone, glycyrrhizic acid, dexamethasone and deoxycorticosterone in repetitive stress. AB - 1. The present study examined the effect of naloxone (NAL), glycyrrhizic acid (GCA), deoxycorticosterone (DOC) and dexamethasone (DEX) on daily repeated 2 h chronic restrained stress (RS) on the locomotor activity (LA) of rats tested in the open field arena to elucidate the possible roles of opioids, glucocorticoids and mineralocorticoids in response to stress. 2. Intact and adrenalectomized (ADX) rats were either injected with 0.1 mL of NAL (0.32 microgram/100 g BW), 2.4 mg/kg DOC or 120 micrograms/kg DEX or had 1.0 mg/mL GCA dissolved in their drinking water or normal saline (for the ADX group) dissolved in their drinking water. 3. In intact groups, treatment with NAL completely blocked the stress response and treatment with GCA, DOC and DEX partially prevented the stress response. Adaptation occurred on either days 4, 5, 6 or 7 for intact rats treated with DEX, DOC, GCA or control rats, respectively. All ADX control rats died following the first 2 h RS. Adrenalectomized rats treated with DEX or DOC adapted later compared with intact rats, while rats given either GCA or NAL were unable to block or adapt to chronic RS. 4. These findings demonstrate that the stress response is primarily mediated by endogenous opioids, in that it is blocked by NAL. Both mineralocorticoids and glucocorticoids, which can act centrally to inhibit endorphins, partially blocked the stress response. The effect of GCA in intact rats was similar to that of both DEX and DOC in intact rats. Adrenalectomized rats treated with GCA (despite their lack of endogenous corticosterone) showed a stress response that was significantly different from the other ADX groups, implying that GCA had effects independent of endogenous corticosterone. PMID- 10386235 TI - Examination of adenosine receptor-mediated relaxation of the pig coronary artery. AB - 1. The adenosine receptors mediating relaxation of porcine isolated left anterior descending coronary arteries (LAD) and the effects of the level and type of preconstriction on the responses to adenosine analogues were examined in the present study. 2. Relaxation responses to the non-selective adenosine receptor agonist N-ethylcarboxamidoadenosine (NECA) were endothelium independent. N Ethylcarboxamidoadenosine, GR 79236 (A1 receptor selective) and 8-cyclopentyl-1,3 dipropylxanthine (CGS 21680) (A2A receptor selective) produced full relaxation in LAD precontracted to 50% of the response to potassium depolarization with the thromboxane receptor agonist U46619. The order of potency was CGS 21680 = NECA > GR 79236, consistent with that defining the A2A receptor subtype. 3. 3,7-Dimethyl 1-propargylxanthine (DMPX; A2 receptor selective) competitively antagonized NECA and CGS 21680 with pKB values of 4.95 +/- 0.09 and 5.06 +/- 0.22, respectively. The A1 receptor selective antagonist 1,3-[3H]-dipropyl-8-cyclopentylxanthine (DPCPX) had no effect on NECA relaxation, even in the presence of DMPX. 4. The sensitivity to relaxation by NECA was dependent on the precontracting agent. Arteries precontracted with endothelin (ET)-1 were most sensitive to NECA, U46619 precontracted arteries were intermediate and KCl-precontracted arteries were least sensitive. 5. The potency of NECA was reduced when the preconstriction level was increased from 50 to 90% of maximum in U46619-precontracted arteries (pEC50 7.94 +/- 0.12 and 7.35 +/- 0.04, respectively) and, in KCl-precontracted arteries, both the potency and maximum effect of NECA were reduced when the preconstriction level increased from 50 to 80% of maximum (pEC50 7.52 +/- 0.13 and 6.91 +/- 0.26, respectively; maximum responses 82.5 +/- 10.2 and 23.9 +/- 3.6%, respectively, of the preconstricted tone). Relaxation responses to NECA were independent of the level of precontraction in ET-1-precontracted arteries. 6. In porcine LAD, relaxation responses to adenosine analogues were endothelium independent and were mediated via A2A adenosine receptors. Responses to NECA were dependent on both the level and type of preconstriction. PMID- 10386236 TI - Short- and long-term effects of glycyrrhizic acid in repetitive stress. AB - 1. This study was carried out to determine the effect of short-term and long-term ingestion of glycyrrhizic acid on the response to 2 h of restraint stress by measuring locomotor activity and plasma corticosterone levels. 2. Male Sprague Dawley rats were randomly assigned into four groups, each group having eight rats. Group 1 (control) was given ordinary tap water, while groups 2 (short term), 3 and 4 (both long term) were given tap water containing 1 mg/mL glycyrrhizic acid to drink for 10 days, 4 weeks and 9 weeks, respectively. All the rats were subjected to 2 h of restraint stress and the locomotor activity assessed using an activity test in an open field arena followed by blood sampling to determine the plasma corticosterone level. These procedures were repeated daily for 14 days. 3. The basal locomotor activity scores for rats given glycyrrhizic acid for 10 days or 4 weeks were similar to those of controls; however, that of the rats treated long term with glycyrrhizic acid was significantly lower (21.0 +/- 3.0 squares crossed; P < 0.0005). Following the first period of restraint stress there was a highly significant decrease in locomotor activity, which remained significantly lower until the seventh and subsequent periods, indicating an adaptation to the repeated stress had occurred. Although the decrease in locomotor activity was partially blocked and adaptation to repetitive stress was enhanced in the rats given glycyrrhizic acid for 10 days, this was not seen in rats treated with glycyrrhizic acid for 4 or 9 weeks. The corticosterone levels in control rats were significantly elevated for 4-5 days following the exposure to repetitive stress but decreased gradually from day 7 onwards. However, both short- and long-term glycyrrhizic acid-treated rats had higher plasma corticosterone levels than the controls (P < 0.05). 4. In conclusion, repetitive restraint stress caused decreased locomotor activity associated with increased plasma corticosterone levels, both of which, in normal rats, decreased with adaptation to stress. The stress response was partially blocked and adaptation enhanced in rats given glycyrrhizic acid for 10 days, but not in rats given glycyrrhizic acid for 4 and 9 weeks. Glycyrrhizic acid ingestion caused high plasma corticosterone. PMID- 10386237 TI - Pertussis toxin-sensitive G-proteins and regulation of blood pressure in the spontaneously hypertensive rat. AB - 1. Increased Gi-protein-mediated receptor-effector coupling in the vasculature of the spontaneously hypertensive rat (SHR) has been proposed as a contributing factor in the maintenance of elevated blood pressure. If increased Gi-protein mediated activity plays an important role in hypertension in SHR, then inhibition of Gi-proteins by pertussis toxin would be expected to decrease blood pressure in this genetic hypertensive model. To address this hypothesis, studies were undertaken comparing the cardiovascular effects of pertussis toxin in SHR and normotensive Wistar-Kyoto (WKY) rats. 2. Spontaneously hypertensive and WKY rats were instrumented with radiotelemetry devices and blood pressure measurements were recorded in conscious rats. Following a single injection of pertussis toxin (10 micrograms/kg, i.v.), mean arterial blood pressure fell from 161 +/- 3 to 146 +/- 1 mmHg in the SHR and the effect was sustained for more than 2 weeks. In contrast, 10 micrograms/kg, i.v., pertussis toxin produced no significant effect on blood pressure in WKY rats (103 +/- 4 vs 101 +/- 5 mmHg). 3. In a separate study, SHR and WKY rats were administered 30 micrograms/kg, i.v., pertussis toxin or 150 microL/kg, i.v., saline and, 3-5 days later, rats were anaesthetized and instrumented to permit measurement of blood pressure and renal function. At this higher dose, pertussis toxin reduced blood pressure in both strains of rat, although the effect was markedly greater in SHR (approximately 40 mmHg decrease) compared with WKY rats (approximately 15 mmHg decrease). In SHR, pertussis toxin increased renal blood flow (from 5.7 +/- 0.3 to 7.5 +/- 0.8 mL/min per g kidney) and decreased renal vascular resistance (from 31 +/- 2 to 19 +/- 2 mmHg/mL per min per g kidney). In WKY rats, pertussis toxin had no significant effect on renal parameters. 4. Results from these studies indicate that a pertussis toxin sensitive Gi-protein-mediated pathway contributes to the maintenance of hypertension and elevated renal vascular tone in the SHR. PMID- 10386238 TI - Bradykinin-induced vascular responses in dog isolated lingual artery. AB - 1. Kinin-induced vascular responses were studied and kinin receptor subtypes were characterized in canine isolated and preconstricted lingual arteries. 2. A low dose of bradykinin (BK; < 3 x 10(-14) mol) induced only vasodilation, while a higher dose of BK (> 3 x 10(-13) mol) frequently induced a biphasic response: a transient constriction followed by dilation. 3. The BK-induced vasodilation was mostly endothelium dependent but was also partly endothelium independent because although the dilation response was greatly reduced after removal of the endothelium, it was not completely abolished. 4. The dilation response to BK was significantly inhibited by the B2 kinin receptor antagonist HOE 140 and was partly reduced by indomethacin (10 mumol/L) (P < 0.05). 5. Bradykinin-induced vasoconstriction was enhanced in endothelium-denuded preparations. The constriction was significantly inhibited by HOE 140 (10(-10) mol/L). The BK induced responses were not affected by the B1 kinin receptor antagonist des-Arg9 [Leu8]-BK (3 x 10(-11) mol/L). 6. The B1 kinin receptor agonist des-Arg9-BK (> 10(-12) mol/L) produced vasodilation in 60% of endothelium-intact preparations. In 20% of the endothelium-intact preparations des-Arg9-BK produced a biphasic response: weak vasoconstriction followed by weak vasodilation. The des-Arg9-BK induced dilation and constriction were significantly inhibited by des-Arg9-[Leu8] BK (3 x 10(-11) mol/L), but were not affected by HOE 140 (10(-10) mol/L). 7. In conclusion, it appears that both B1 and B2 kinin receptors are present in the dog lingual artery. Both receptor subtypes mediate either vasodilation or vasoconstriction and BK-induced vasodilation is mostly endothelium dependent, although it may also be partially prostaglandin dependent. PMID- 10386239 TI - The 5-HT2 receptor antagonist sarpogrelate reduces urinary and plasma levels of thromboxane A2 and urinary albumin excretion in non-insulin-dependent diabetes mellitus patients. AB - 1. Therapeutic effects of a 5-HT2 receptor antagonist sarpogrelate on microalbuminuria and thromboxane (TX)A2 biosynthesis were examined in non-insulin dependent diabetes mellitus (NIDDM) patients. 2. In protocol I, the ankle brachial pressure index (API; an indicator of peripheral blood flow) and urinary albumin excretion (UalbV; an indicator of renal function) were determined in 42 NIDDM patients who had been treated with 300 mg/day sarpogrelate for 8 weeks. In an analysis of the results, the NIDDM patients were divided into four groups based on the severity of either vasculopathy or nephropathy as follows: group A, API < 0.9, UalbV > or = 100 mg/day; group B, API < 0.9, UalbV < 100 mg/day; group CAPI > or = 0.9, UalbV > or = 100 mg/day; and group D, API > or = 0.9, UalbV < 100 mg/day. 3. In protocol II, 10 NIDDM patients with UalbV values > 100 mg/day were divided into two groups to further confirm the effect of sarpogrelate on albuminuria: group E, the sarpogrelate treatment group (n = 5); and group F, the no treatment group (n = 5). 4. In protocol I, the incidence of a cold sensation in the lower extremities was reduced from 45.2 to 21.4% following sarpogrelate treatment. In patients with UalbV > or = 100 mg/day (groups A and C), UalbV was significantly decreased independent of API, while it did not change in patients with UalbV < 100 mg/day (groups B and D). Plasma TXB2 levels were significantly decreased following sarpogrelate treatment, whereas plasma 6-keto-prostaglandin F1 alpha levels were not. 5. In protocol II, in the sarpogrelate treatment group (group E), albuminuria was significantly improved and both plasma levels TXB2 and urinary TXB2 excretion were significantly decreased. In contrast, in the untreated group (group F), neither plasma levels TXB2 nor urinary TXB2 excretion was changed. 6. In conclusion, microalbuminuria was improved by treatment with the 5-HT2 receptor antagonist sarpogrelate independent of latent vasculopathy. Blockade of 5-HT2 receptors is suggested to be beneficial for the treatment of nephropathy in NIDDM patients. It is possible that the inhibition of TXA2 biosynthesis is involved in the therapeutic effect of 5-HT2 receptor antagonists. PMID- 10386240 TI - Both extracellular ATP and shear stress regulate the release of nitric oxide in rat caudal artery. AB - 1. To elucidate the physiological role of nitric oxide (NO) in regulating vascular tone, the effects of NG-nitro-L-arginine methyl ester (L-NAME), an NO synthase inhibitor, on the vasoconstrictor response to noradrenaline (NA) in rat caudal artery was examined. 2. NG-Nitro-L-arginine methyl ester significantly potentiated the NA-induced increase in perfusion pressure in the perfused caudal artery, but did not affect the NA-induced contraction in caudal artery ring preparations. In addition, an increase in perfusion pressure mechanically produced by a stepwise increase in flow rate was not affected by L-NAME. 3. Noradrenaline evoked a significant increase in the release of endogenous ATP and its metabolites from the perfused artery, whereas increased perfusion pressure as a result of increased flow rate did not evoke release of endogenous ATP. 4. In the presence of exogenously applied ATP, L-NAME significantly potentiated the increase in perfusion pressure produced by increased flow rate. 5. These results indicate that perfused vascular tone is regulated by endogenous NO and suggest that extracellular ATP may participate in the synthesis and release of NO by shear stress in endothelial cells in the rat caudal artery. PMID- 10386241 TI - Role of nitric oxide and endothelium in the flow-induced dilation of rat coronary arteries under two preconstriction conditions. AB - 1. Pressure-induced tone and flow-induced dilations were studied in a rat perfused epicardial coronary artery mounted in an arteriograph. Spontaneous tone was assessed in arteries submitted either to 60 or 90 mmHg intraluminal pressure either under control conditions, after incubation with NG-nitro-L-arginine methyl ester (L-NAME; 100 mumol/L) or after endothelial denudation. Flow-induced dilation was quantified under these conditions in preparations either submitted to 60 mmHg and preconstricted with 10 mumol/L 5-hydroxytryptamine (5-HT) or exhibiting spontaneous tone at 90 mmHg. 2. Spontaneous tone was greater at 90 mmHg compared with tone obtained at 60 mmHg (21 +/- 2 vs 10 +/- 2% reduction of the fully dilated diameter after sodium nitroprusside incubation, respectively). Incubation with L-NAME or removal of the endothelium significantly increased spontaneous tone at both pressures compared with control. 3. In arteries submitted to 60 mmHg and preconstricted with 10 mumol/L 5-HT, flow (0-800 microL/min) induced a continuous dilation (maximal value 63 +/- 4%). As a function of flow, shear stress first increased and then plateaued at values of approximately 76 +/- 6 dyn/cm2. After L-NAME incubation or endothelial denudation, the flow-induced dilation was reduced to the same extent and was obtained for higher values of shear stress (172 +/- 14 and 150 +/- 14 dyn/cm2, respectively). 4. In arteries exhibiting spontaneous tone, starting flow led, first, to a constriction followed by a dilation up to 76 +/- 4% of the initial tone. Incubation with L-NAME greatly altered flow-induced dilation. Endothelium removal further reduced the dilation obtained for very high values of shear stress (up to 300 dyn/cm2). 5. The present study shows that different patterns of vasodilation induced by flow can be observed, depending on the initial vasoconstrictor stimulus. In 5-HT-preconstricted arteries, flow-induced dilation appears to be fully dependent on the synthesis and release of nitric oxide. In arteries with spontaneous tone, a vasoconstrictor substance could be released for low values of flow. Nitric oxide is mainly, but not exclusively, responsible for the vasodilation. For both experimental conditions, removal of the endothelium greatly reduced the response, but a dilation was still observed. PMID- 10386242 TI - Comparison of oscillometric blood pressure measurements at the wrist with an upper-arm auscultatory mercury sphygmomanometer. AB - 1. Oscillometric devices for blood pressure (BP) measurement at the wrist are becoming more widely used in clinical practice. However, systematic comparisons with standard auscultatory BP measurement at the brachial artery are scarce. Therefore, we compared two such devices, the Boso-Mediwatch (Bosch & Sohn GmbH U. Co., Jungingen, Germany) and the Omron R3 (Omron Corp., Tokyo, Japan), with upper arm auscultatory mercury sphygmomanometry. 2. In 20 normotensive subjects and 20 treated hypertensive subjects, the Boso-Mediwatch was applied to the left wrist by observer 1 and was compared with mercury sphygmomanometry of the right upper arm by observer 2. Each observer swapped sides and the procedure was repeated. The Boso-Mediwatch was then applied to the right wrist by observer 1 and was compared with mercury sphygmomanometry of the left upper arm by observer 2. Each observer once again swapped sides and the procedure was repeated. An identical protocol was followed for the Omron R3 in a further 20 treated hypertensive subjects and 20 normotensive subjects. 3. There were no significant differences between observers or left versus right arm for either oscillometric device or when measurements were performed by mercury sphygmomanometry. In normotensive subjects, the Boso-Mediwatch readings were higher than mercury sphygmomanometer readings, with mean differences (95% confidence intervals) of 3.9 (0.1, 7.6; P = 0.045) and 7.0 mmHg (4.7, 9.2; P < 0.001) for systolic and diastolic BP, respectively. In hypertensive subjects, the Boso-Mediwatch readings were lower for systolic BP (mean difference -6.0 mmHg (-11.6, -0.3; P = 0.04) but were higher for diastolic BP (mean difference 3.8 mmHg (1.4, 6.3; P < 0.01). 4. In normotensive subjects, the Omron R3 readings were higher, with mean differences of 3.2 (0.6, 5.8; P = 0.018) and 4.2 mmHg (1.6, 6.7; P = 0.003) for systolic and diastolic BP, respectively. In hypertensive subjects, the Omron R3 readings were lower for both systolic and diastolic BP (mean differences: -5.8 (-8.8, -2.8; P = 0.001) and -5.5 (-9.3, -1.6; P = 0.008), respectively). 5. For the Boso Mediwatch, weighted least products analysis confirmed the presence of both fixed and proportional error for systolic BP but not for diastolic BP. For the Omron R3, fixed or proportional error was not detected for either systolic or diastolic BP. 6. These wrist oscillometric devices, although offering portability and convenience, give BP measurements that frequently differ substantially (by at least 5 mmHg) from readings simultaneously measured at the upper arm by a mercury sphygmomanometer. The magnitude and direction of differences detected are dependent on both the device used and the underlying level of BP. PMID- 10386243 TI - Increased oxidative DNA damage in stroke-prone spontaneously hypertensive rats. AB - 1. The amount of urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG), a biomarker of the total systemic oxidative stress in vivo, in stroke-prone spontaneously hypertensive rats (SHRSP) was not different from that in control normotensive Wistar-Kyoto (WKY) rats at 6 weeks of age, but became higher than control values after the development of severe hypertension at 14-17 weeks of age. 2. The amount of urinary 8-OHdG was not significantly different between SHRSP treated with anti hypertensive agents and those not treated at 14 weeks of age. 3. Stroke-prone spontaneously hypertensive rats were exposed to DNA damage by oxidative stress at the early stage when they developed severe hypertension, but this increase in DNA damage was not the secondary effect of hypertension. PMID- 10386244 TI - Cerebrovascular contractility after clot removal in monkey subarachnoid haemorrhage. AB - 1. The effects of mechanical clot removal during early surgery on pharmacological cerebrovascular reactivity after subarachnoid haemorrhage (SAH) were investigated in the monkey. 2. Contractions to potassium chloride, 5-hydroxytryptamine and noradrenaline in rings of proximal parts of middle cerebral arteries (MCP), surrounded with clot, and basilar arteries (BAP), far from the clot, were examined 7 days after SAH, in which an autologous blood clot was bilaterally placed around major cerebral arteries. 3. Compared with the sham-operated group, contractions in the clot removal groups at 48 and 72 h after SAH were reduced in MCP and enhanced in BAP. 4. These results suggest that divergent vascular contractility may occur according to the distance between artery and clot if the clot is removed later than 48 h after SAH. PMID- 10386245 TI - Transcriptional control by corticosteroids of CHIF gene expression in the rat distal colon. AB - 1. Previous studies have shown that levels of CHIF mRNA are increased in the distal colon of the rat in response to corticosteroids. We have recently reported that this response occurs within 2 h of a single dose of either dexamethasone or aldosterone and that the response is mediated via both the mineralocorticoid and glucocorticoid receptors. 2. In the present study we sought to further define the nature of the various transcripts detected by a CHIF coding region cRNA probe in northern blot analysis of corticosteroid-stimulated colonic RNA. The identification of an intronic sequence was used to synthesize an intron-specific cDNA probe to characterize the transcripts. 3. The presence of an intronic sequence in the originally published sequence was confirmed using coupled reverse transcriptase-polymerase chain reactions with primers spanning and within the intronic sequence. The intronic cDNA probe hybridized to the higher molecular weight transcripts detected by the cRNA probe. These transcripts are induced in response to both corticosteroids. 4. Taken together with our observations that the increase in CHIF mRNA levels in the distal colon in response to corticosteroids is not blocked by prior cycloheximide treatment, the increase in the levels of the primary transcript and partially spliced forms argues that this is a primary transcriptional response. This is the first clear demonstration of an aldosterone-induced gene in vivo in a mammalian system. PMID- 10386246 TI - The moving finger writes ... 20 years of peripheral dopamine research. PMID- 10386247 TI - Agonism and inverse agonism at dopamine D2-like receptors. AB - 1. The processes that follow the binding of ligands to receptors are critical for their physiological functions. In the present paper I intend to review our own work and the work of other laboratories attempting to understand these processes for the dopamine D2-like receptors (D2, D3, D4) and how they contribute to the mechanisms of drug action. It is thought that the key event in agonist action for these receptors is the stabilization, by the agonist, of the agonist-receptor-G protein ternary complex. The majority of the work I shall describe has been performed using recombinant receptors expressed in cell lines and the mechanisms of receptor action have been probed using ligand binding (competition vs [3H] spiperone), the stimulation of [35S]-GTP gamma S binding and inhibition of adenylyl cyclase. 2. Measures of the ability of agonists to stabilize the agonist receptor-G-protein ternary complex may be obtained in ligand-binding studies using the ratio of dissociation constants for the higher and lower affinity states (KI/KH ratio). The stimulation of [35S]-GTP gamma S binding provides a very convenient assay for agonist action and allows the determination of agonist potency and maximal response. Estimates of these quantities may also be obtained from the inhibition of adenylyl cyclase. For a range of agonists at the D2 receptor, there is a tendency for high values of KI/KH to predict high maximal activity and vice versa, but there is no general correlation. This suggests that the simple scheme of agonist action depending on the stabilization of the ternary complex is an over-simplification and further efficacy determining steps need to be included. For a number of receptors, including the D2 and D3 receptors, it has now been shown that there is activity in the absence of agonist (so-called constitutive activity). This agonist-independent activity can be inhibited by compounds previously considered to be antagonists (e.g. the antipsychotic drugs). Therefore, these compounds are inverse agonists rather than antagonists. The mechanism of this inverse agonist effect is unclear and we are examining this using a variety of biochemical approaches, including the use of constitutively active mutants. 3. The mechanisms of agonism and inverse agonism may be probed using biochemical assays and these studies are of great relevance to the understanding of drug action. PMID- 10386248 TI - Role of dopamine in the pathogenesis of hypertension. AB - 1. Dopamine, via different dopamine receptor subtypes, regulates cardiovascular functions by actions on the central and peripheral nervous systems, vascular smooth muscle, the heart and the kidney. The dopaminergic system in the central nervous system (CNS) may participate in the regulation of systemic blood pressure. 2. Dopamine 'D2-like' (D2, D3 and D4) receptors, rather than 'D1-like' (D1 and D5) receptors, are involved in the CNS regulation of blood pressure; post synaptic D2-like receptors increase blood pressure, while presynaptic D2-like receptors (the predominant action) produce the opposite effect. 3. Outside the CNS, dopamine may regulate blood pressure via pressure controls that act with intermediate rapidity (e.g. stress relaxation, arginine vasopressin and renin angiotensin vasoconstriction), as well as those systems related to the long-term control of body fluid volume. 4. Dopamine D1- and D2-like receptors have been described in resistance vessels, such as the renal, mesenteric, coronary, pulmonary and cerebral arteries. The ability of D1-like receptors to inhibit renal smooth muscle hypertrophy indicates their importance in longer-term regulation of blood pressure. 5. Aberrant dopaminergic regulation of aldosterone secretion, via D2-like receptors, has been reported to be involved in some forms of hyperaldosteronism and hypertension. Some forms of hypertension may also be caused by an aberrant renal dopaminergic system. Abnormalities of three aspects of the renal dopaminergic system may lead to hypertension: (i) renal production of dopamine; (ii) transduction of the renal vascular dopamine signal; and (iii) transduction of the renal tubular dopamine signal. 6. Thus, increased blood pressure occurs after either blockade of D1-like receptors or of dopamine production in rats or disruption of the D1 receptor or the D3 receptor gene in mice. PMID- 10386249 TI - Non-neuronal dopamine in the gastrointestinal system. AB - 1. Dopamine (DA) is a protective agent in the gastrointestinal (GI) tract in both rats and humans. Therefore, we have studied the site of DA production in rat and human GI tract using a variety of techniques, including immunocytochemistry (ICC), in situ hybridization histochemistry, reverse transcription-polymerase chain reaction, HPLC, western blotting and immunoelectron microscopy. 2. We found very high concentrations of DA that persisted after chemical sympathectomy (CS) in the gastric juice, the stomach mucosa and in the pancreas. Both the stomach mucosa and the pancreas also had tyrosine hydroxylase (TH) activity, most of which remained after CS. Double-labelling ICC showed that acid-producing parietal cells and the exocrine pancreas must also be capable of producing DA. 3. We isolated rat stomach parietal cells by cell fractionation and found that both DA and TH activity are present in isolated (denervated) parietal cells. These cells also have other features of aminergic cells: they are immuno- (and mRNA) positive for the DA plasma membrane transporter and vesicular monoamine transporter(s). In both gastric and duodenal mucosa, we demonstrated the presence of significant amounts of the D5 receptor that could serve as a target for locally produced DA. 4. Because DA, its biosynthetic enzymes and its transporters are also found in parietal cells in the human stomach, a mucosal protective system involving DA could be important clinically. PMID- 10386250 TI - Renal-dose (low-dose) dopamine for the treatment of sepsis-related and other forms of acute renal failure: ineffective and probably dangerous. AB - 1. Low-dose ('renal-dose') dopamine (i.e. 1-3 micrograms/kg per min) is used widely for the treatment of acute renal failure induced by ischaemia, toxins and/or sepsis. Here we review the scientific rationale, experimental studies and clinical trials evaluating its use in these settings. 2. Renal-dose dopamine augments renal blood flow, sodium excretion and probably glomerular filtration rate in healthy humans and experimental animals and limits ATP utilization and oxygen requirements in nephron segments at risk of ischaemic injury. Renal-dose dopamine is renoprotective in several ischaemic and nephrotoxic models of acute renal failure. 3. However, most studies in humans have not demonstrated prevention of acute renal failure in high-risk patients or improved outcome in those with established acute renal failure. While the safety profile of dopamine in these settings has not been extensively defined, it is known the drug may precipitate serious cardiovascular and metabolic complications in the critically ill. Therefore, we suggest that renal-dose dopamine should not be used for selective renal vasodilatory and natriuretic actions in those patients with acute renal failure until its efficacy is established in randomized control trials. 4. Renal-dose dopamine may be most valuable when combined with agents targeting other events in acute renal failure, such as cast formation, epithelial cell injury and tubule regeneration. These recommendations should not preclude the use of dopamine for its systemic effects in heart failure and septic shock. PMID- 10386251 TI - Physiological importance of dopamine as a noradrenaline precursor in the corpus luteum. AB - 1. Both in vivo and in vitro studies have demonstrated that the adrenergic innervation of the ovary affects corpus luteum (CL) secretory function in many species. 2. In cattle, ovarian noradrenergic stimulation or the administration of noradrenaline (NA) to the ovary increases ovarian oxytocin (OT) secretion and post-translational processing of OT synthesis within a few minutes. Furthermore, NA affects both progesterone release and its synthesis by increasing cytochrome P450sec and 3 beta-hydroxysteroid dehydrogenase activity. This effect is mediated via luteal cell beta 1- and beta 2-adrenoceptors. 3. The total number of luteal beta-adrenoceptors correlates with peripheral progesterone concentrations during the luteal phase. Conversely, ovarian denervation causes a decrease of steroidogenic activity in the corpus luteum, an increase in beta-adrenoceptors on luteal cells, a delay in follicular development and the disruption of cyclicity, as well as other effects. Moreover, brief pharmacological blockade of ovarian beta-adrenoceptors in the mid-cycle of cattle decreases progesterone secretion by 20-40%. 4. We conclude that tonic beta-adrenoceptor stimulation of the CL ensures the basal secretion of progesterone, whereas acute noradrenergic activation supports the CL during stressful situations that could impair its function. Dopamine concentrations within the CL are highly correlated with those of NA during the oestrous cycle and are higher in the newly formed compared with the developed CL, the regressed CL or the CL of pregnant females. PMID- 10386252 TI - Anti-atherosclerotic action of vascular D1 receptors. AB - 1. Vascular smooth muscle cell (VSMC) migration and proliferation are believed to play key roles in atherosclerosis. To elucidate the role of vascular dopamine D1 like (D1 and D5) receptors in atherosclerosis, the effects of dopamine and the specific D1-like receptor agonists SKF 38393 and YM 435 on platelet-derived growth factor (PDGF)-BB-mediated VSMC migration, proliferation and hypertrophy were investigated. 2. We observed that cell stimulated by 5 ng/mL PDGF-BB showed increased migration, proliferation and hypertrophy. These effects were prevented by co-incubation with dopamine, SKF 38393 or YM 435 at 1-10 mumol/L and this prevention was reversed by Sch 23390 (1-10 mumol/L), a specific D1-like receptor antagonist. These actions of D1-like receptor agonists were mimicked by 1-10 mumol/L forskolin, a direct activator of adenylate cyclase, and 0.1-1 mmol/L 8 bromo-cAMP. The actions were blocked by the specific protein kinase A (PKA) inhibitor N-[2-(p-bromocinnamylamino) ethyl]-5-isoquinoline-sulphonamide (H 89), but were not blocked by its negative control N-[2-(N-formyl-p chlorocinnamylamino) ethyl]-5-isoquinoline sulphonamide (H 85). Platelet-derived growth factor-BB (5 ng/mL)-mediated activation of phospholipase D (PLD), protein kinase C (PKC) and mitogen-activated protein kinase (MAPK) activity was significantly suppressed by co-incubation with dopamine. 3. These results suggest that vascular D1-like receptor agonists inhibit migration, proliferation and hypertrophy of VSMC, possibly through the activation of PKA and the suppression of activated PLD, PKC and MAPK activity. PMID- 10386253 TI - Dopamine sulphate: an enigma resolved. AB - 1. The source and physiological significance of dopamine (DA) sulphate, which exists in plasma at much higher concentrations than free DA, have long been a puzzle. The present article reviews how the convergence of modern molecular and traditional clinical approaches is shedding new light on the origins and meaning of DA sulphate. 2. The sulphotransferase isoenzyme responsible for production of DA sulphate in humans (SULT1A3) has been cloned and shown to be expressed in large quantities in the gastro-intestinal tract, but not in liver. No orthologue of SULT1A3 has yet been identified in other species, consistent with the greater importance of sulphate conjugation of DA in humans than in most animals. 3. Diet has a major impact on plasma DA sulphate, with dramatic increases after ingestion of meals and foods rich in biogenic amines; however, substantial amounts of DA sulphate remaining after prolonged fasting indicate the presence of a mainly endogenous source. The lack of influence of acute or chronic changes in sympathetic outflow or of sympathoneural degeneration on plasma DA sulphate indicates that DA sulphate does not derive from sympathetic nerve. Relatively low rates of production from intravenously infused DA indicate that very little DA sulphate (< 2%) derives from metabolism of circulating DA, such as in red cells or platelets. 4. Consistent increments in DA sulphate from arterial to the outflowing venous plasma draining mesenteric organs, without increments across other organs or tissues (e.g., heart, lungs, liver), indicate that the gastrointestinal tract is a major source of more than 75% of DA sulphate produced in the body. The gastro-intestinal tract is also the site of a novel DA autocrine/paracrine system that produces nearly 50% of the DA in the body. Therefore, production of DA sulphate appears to reflect an enzymatic 'gut-blood' barrier for detoxifying dietary biogenic amines and delimiting autocrine/paracrine effects of endogenous DA generated in a novel 'third catecholamine system'. PMID- 10386254 TI - Pulmonary complications of mechanical ventilation. AB - Although life-saving, mechanical ventilation may be associated with many complications, including consequences of positive intrathoracic pressure, the many aspects of volutrauma, and adverse effects of intubation and tracheostomy. Optimal ventilatory care requires implementing mechanical ventilation with attention to minimizing adverse hemodynamic effects, averting volutrauma, and effecting freedom from mechanical ventilation as quickly as possible so as to minimize the risk of airway complications. PMID- 10386255 TI - Complications of hemodynamic monitoring. AB - The uses of the pulmonary artery catheter have been expanded from its original use, helping to assess the cardiac output and left ventricular filling pressure of patients with cardiac disease, to include the management of patients with trauma, septic shock, respiratory failure, and those undergoing high-risk surgeries. Although more than 1 million pulmonary artery catheters are inserted each year in the United States, clear evidence establishing that they improve outcome remains hard to find. This article discusses the complications of invasive hemodynamic monitoring. PMID- 10386256 TI - Cardiac complications in the intensive care unit. AB - Advances in the care of critically ill patients has been startling, especially in patients with acute coronary syndromes. With new therapies and procedures, however, have come new complications. On balance, our patients are better off, but the stakes are now higher and the complications more serious. The need for constant vigilance has never been greater. PMID- 10386257 TI - Respiratory infectious complications in the intensive care unit. AB - Ventilator-associated pneumonia is the most common infectious respiratory complication in intensive care unit patients, particularly those needing mechanical ventilation. Ventilator-associated pneumonia represents a challenging problem in terms of diagnosis, treatment, and prevention. Nosocomial sinusitis is another respiratory infection, not uncommon in mechanically ventilated patients. This type of infection has to be suspected in nasally intubated patients and may be a hidden focus of fever and sepsis. PMID- 10386258 TI - Antimicrobial resistance in intensive care units. AB - The unique nature of the intensive care unit (ICU) environment makes this part of the hospital a focus for the emergence and spread of many antimicrobial-resistant pathogens. There are ample opportunities for the cross-transmission of resistant bacteria from patient to patient, and patients are commonly exposed to broad spectrum antimicrobial agents. Rates of resistance have increased for most pathogens associated with nosocomial infections among ICU patients, and rates are almost universally higher among ICU patients compared with non-ICU patients. There are many opportunities, however, to prevent the emergence and spread of these resistant pathogens through improved use of established infection control measures (i.e., patient isolation, hand washing, glove use, and appropriate gown use), and implementation of a systematic review of antimicrobial use. PMID- 10386259 TI - Pleural complications in the intensive care unit. AB - In summary pleural complications in the ICU are common. Pneumothorax in a mechanically ventilated patient is a medical emergency that requires prompt diagnosis and therapy. Correct diagnosis and therapy of pleural effusions will assist in improving pulmonary physiology and outcome in the ICU patient. PMID- 10386260 TI - Gastrointestinal complications in the intensive care unit. AB - Pathologic conditions affecting the abdomen are a significant cause of morbidity and mortality in the intensive care unit, but their importance is not widely recognized. This article presents several aspects of abdominal pathology that can occur in intensive care unit patients. This pathology may have a considerable impact on the prognosis and survival of the critically ill patient. The diagnostic contribution of laboratory tests and imaging is discussed. Conditions such as the abdominal compartment syndrome, acute mesenteric ischemia, gastrointestinal bleeding, diarrhea, abdominal sepsis, complications of entereal and parenteral nutrition, and ileus in critically ill patients are also reviewed. PMID- 10386261 TI - Acute renal failure in the intensive care unit. Therapy overview, patient risk stratification, complications of renal replacement, and special circumstances. AB - This article provides a basic definition of severity scoring among patients with acute renal failure and extends the definition into the types of dialysis support that are generally used in intensive care unit acute renal failure. Acute dialysis dosing and the problems that create a difference between chronic renal failure and acute renal failure support are described, the dialytic techniques and side effects and complications of each are compared, and nonrenal-based special situations in which extracorporeal therapy has been found to be helpful are defined. PMID- 10386262 TI - Venous thromboembolism in the intensive care unit. AB - Venous thromboembolic disease has emerged as a significant cause of morbidity and mortality in hospitalized patients. This article reviews the salient features of venous thromboembolism as they pertain to the critically ill. Emphasis is placed on identifying risk factors, diagnostic strategies, prophylaxis, and treatment of this disorder. Deep venous thrombosis and pulmonary embolism, both being manifestations of the same disease processes, are considered together in this discussion of venous thromboembolism. PMID- 10386263 TI - Drug interactions in the intensive care unit. AB - Adverse drug reactions are a major source of complications in the intensive care unit. Drug interactions contribute significantly to the incidence of adverse drug reactions. The intensive care unit clinician must remain aware of the major mechanisms for drug interactions, which are reviewed in this article. PMID- 10386264 TI - Endocrinologic and metabolic complications in the intensive care unit. AB - Critical illness provides major stresses on all body systems, including those serving important regulatory functions. Endocrinologic and metabolic abnormalities are common on presentation and during hospitalization in the intensive care unit. Some of these abnormalities are the focus of this article. The authors review abnormalities of the adrenal and thyroid glands and in the metabolism of glucose, and include a brief review of abnormalities of sodium and calcium metabolism. PMID- 10386265 TI - Neurologic complications in the intensive care unit. AB - Neurologic complications resulting from critical illness and intensive care unit therapies are common, but frequently unrecognized because these patients are often intubated, sedated, and, occasionally, receiving neuromuscular blocking agents. Neurologic complications are associated with an increased intensive care unit mortality. This article discusses central nervous system complications that are secondary to critical illness or to therapeutic interventions in the critically ill patient. PMID- 10386266 TI - Muscle dysfunction in the intensive care unit. AB - Muscle weakness, particularly impairment of the respiratory muscles, is a frequent abnormality in ICU patients. This is more relevant in some clinical situations--for example, in weaning patients from mechanical ventilation. Intensive care procedures that are designed to "rest" respiratory muscles, such as mechanical ventilation, may also contribute to impaired muscle function. Pharmacologic administration of glucocorticoids, several antibiotics, NMB agents, and so on has the potential to cause untoward effects. The development of myopathy and prolonged paresis has been increasingly recognized after prolonged use of these drugs in the ICU. Pathophysiologic changes in the nerve, muscle, or neuromuscular junction associated with the patient's underlying condition may also play a role in the development of impaired function. The assessment of muscle function is difficult and inaccurate. The techniques developed have a poor predictive value because of the difficulty in making the measurements in uncooperative patients and the lack of standardization. Furthermore, it is likely that some voluntary maneuvers underestimate muscle strength. Invasive procedures such as phrenic nerve stimulation or EMG recording are also of limited value. PMID- 10386267 TI - Skin complications in the intensive care unit. AB - Critically ill patients usually have multiple risk factors for the development of pressure ulcers. Pressure ulcers involve all levels of tissue from bone to skin, and result from excessive pressure and shearing. Control of incontinence, maintenance of adequate oxygen delivery and nutritional support is the key to minimizing the effects of skin breakdown in the intensive care unit. Consistent assessment and surveillance of skin for early signs of pressure ulcer development are essential, as is an interdisciplinary approach with nursing specialists and surgical consultants for pressure ulcers that have developed. PMID- 10386268 TI - The molecular basis of genetics and inheritance. AB - This article reviews the molecular basis of genetic disorders. It is presented at an introductory level, assuming that the reader has a good physiologic background but has little expertise in the fields of molecular biology and molecular genetics. It addresses the following questions: (1) What is DNA? (2) What are genes and chromosomes? (3) How are genes expressed and how is gene expression regulated? (4) How is DNA replicated? (5) How is genetic material inherited? (6) How is phenotype determined? (7) How are genetic diseases inherited? The goal of this article is to provide vocabulary and concepts that are key for understanding the substantive articles that follow on the subjects of clinical genetics, gene therapy, ethical issues in genetics, and the molecular genetics of cardiovascular disorders. Included in this article is a list of key terms with a corresponding page number where the term is defined or discussed. This should be used as a resource for reading the other articles. In addition, further readings, programmed CDs, and web sites in the areas of molecular biology and molecular genetics are suggested. PMID- 10386269 TI - Clinical genetics: an overview. AB - It is imperative that clinicians understand basic human genetic concepts. In this article, the patterns of inheritance for single gene disorders are described and include the traditional Mendelian, nontraditional, and multifactorial patterns. Next, chromosomal and structural abnormalities are described, followed by a description of the current measures by which genes are detected, including gene mapping and polymorphisms, and concluding with cytogenetic, molecular, and biochemical testing. PMID- 10386271 TI - Familial hypertrophic cardiomyopathy. AB - Familial hypertrophic cardiomyopathy (FHC) is a cardiomyopathy that occurs in 0.2% of the general population. It is characterized by asymmetrical hypertrophy of the ventricle, predominantly the intraventricular septum. FHC is caused by genetic mutations in several of the sarcomeric proteins, such as myosin heavy chain, troponin T, troponin I, alpha-tropomyosin, essential and regulatory light chains of myosin, and the cardiac myosin-binding protein C. FHC is genetically heterogeneous, and, therefore, it is associated with a very diverse clinical presentation in terms of altered cardiac structure and clinical manifestations. The most severe manifestation is sudden death. The purpose of this article is to provide the reader with new insights into the genetic mutations that give rise to FHC and to discuss risk factors that are associated with severe hypertrophy and sudden death in this population. PMID- 10386270 TI - The genetic basis for cardiac dysrhythmias and the long QT syndrome. AB - Cardiac muscle excitation is the result of ion fluxes through cellular membrane channels. Any alterations in channel proteins that produce abnormal ionic fluxes will change the cardiac action potential and the pattern of electrical firing within the heart. The idiopathic long QT syndrome (LQTS) is an inherited cardiac pathology localized to mutated genes encoding for myocardial, voltage-activated sodium and potassium ion channels. The expression of abnormal sodium and potassium channels results in aberrant ionic fluxes that produce a prolonged ventricular repolarization. This prolonged time to repolarization is the electrophysiologic basis for prolongation of the QT interval. Individuals with LQTS are at significant risk for developing lethal ventricular dysrhythmias due to an abnormal pattern of cardiac excitation. Identification of a genetic basis for LQTS has had significant implications for genetic counseling, the development of effective antidysrhythmic drug therapies, and nursing interventions. PMID- 10386272 TI - Genetic determinants of blood pressure regulation. AB - Blood pressure homeostasis in humans reflects the coordinate interactions of cardiac output, peripheral vascular resistance, renal volume control, and CNS integration in response to short- and long-term environmental stimuli. Variations in mean arterial pressure within the population include a significant hereditary component. The clearest examples of this genetic contribution occur in rare forms of monogenic hypertension (glucocorticoid remediable aldosteronism, apparent mineralocoid excess, Liddle's syndrome) or hypotension (pseudohypoaldosteronism type I, Bartter's syndrome, Gitelman's syndrome). Primary hypertension, which comprises approximately 95% of hypertensives and is a major risk factor for coronary heart disease, stroke, and renal disease in the U.S., represents a multifactorial and polygenic disease with incremental contributions from genetic and environmental determinants. Efforts to date have identified several candidate genes involved in primary hypertension, including angiotensinogen (AGT), a vasoactive peptide; alpha-adducin, a protein that regulates sodium transport; and the G protein beta 3 subunit, a protein involved in intracellular signal transduction. Advances in knowledge and technology associated with the Human Genome Project, combined with continuing basic research on the physiologic and biochemical causes of hypertension, offer promise for improved diagnosis and therapy of this prevalent disease. PMID- 10386273 TI - The use of molecular technologies for the detection of enteroviral ribonucleic acid in myocarditis. AB - The introduction of molecular technology to the field of cardiovascular research has revolutionized the diagnosis and determination of the pathogenesis of diseases. This has been the case for viral myocarditis. Although rapid identification and specific treatment for viral disorders such as myocarditis continue to challenge researchers, molecular detection techniques have provided an insight into the role of viral genomes in this disorder. Although in situ hybridization (ISH) continues to be an effective detection method and is utilized in many laboratories, polymerase chain reaction (PCR) techniques are fast becoming the standard for molecular analysis in patients with viral myocarditis. Following a review of viral myocarditis, the roles of ISH and PCR will be discussed. Lastly, clinical relevance and areas of future research will be presented. PMID- 10386274 TI - Gene therapy as a therapeutic intervention for vascular disease. AB - Gene therapy for the treatment of many medical problems, including vascular disease, has become the subject of increasing discussion in both the scientific literature and the national press over the past decade. This review will examine the history and current status of gene therapy for vascular proliferative disorders and advanced chronic peripheral and cardiac ischemia. PMID- 10386275 TI - Genetic testing, screening, and counseling issues in cardiovascular disease. AB - Genetic risk assessment for cardiovascular disease is less advanced and less widely performed to date than it is for cancer. Yet it is no less important. Alert clinicians should "think genetically" and follow up appropriately when confronted with a client having a family history of heart disease, early heart disease themselves, a known genetic disorder in which cardiac problems may be a component, or signs and symptoms indicative of a familial component to the heart problem observed. It is important for the clinician to know how, when, and to whom referral for further genetic evaluation and counseling should be made. Genetic testing and screening in children or adolescents for conditions such as hypertrophic cardiomyopathy (HCM), familial hypercholesterolemia (FH), and long QT (LQT) syndrome, when indicated, can help to save lives through preventive treatment and therapeutic interventions. Preparticipation sports physicals are one means of providing such screening and are important to conduct properly under guidelines recommended by the American Heart Association. Genetic testing for relatives of persons already identified to have heritable cardiac conditions is becoming more and more integral to mainstream primary health care but engender controversy when testing of children is involved. Clinicians must know how to interpret the results of such tests. Appropriate genetic counseling must accompany risk assessment, genetic testing, and screening for cardiovascular disease. PMID- 10386276 TI - Welcome to the Society for Education in Anesthesia. PMID- 10386277 TI - Painless intravenous catheterization by intradermal jet injection of lidocaine: a randomized trial. AB - STUDY OBJECTIVE: To compare efficacy and cost of lidocaine cutaneous anesthesia by two jet injectors to routine needle infiltration for pain relief of intravenous (i.v.) catheterization, hypothesizing that jet injection of lidocaine is less painful than its needle infiltration. DESIGN: Randomized, prospective, controlled trial. SETTING: University hospital outpatient surgical unit. PARTICIPANTS: 75 surgical patients ASA I and II. INTERVENTIONS: Three groups of 25 patients each were given intradermal lidocaine anesthesia via conventional 25 gauge needle/syringe; by MedEJet or Biojector jet injector prior to IV catheterization with an 18-gauge Jelco catheter. MEASUREMENTS AND MAIN RESULTS: Visual analogue pain scores (VAS) (0 = no pain, 10 = intolerable pain) and subjective pain intensity scores (PIS) (0 = not painful, 4 = intolerable pain) at lidocaine application and at i.v. catheterization, were recorded. Cost assessment of each method was made. At local anesthetic application, no pain by proportion of VAS = 0 with MedEJet: 25/25 (confidence interval [CI]: 0.868, 0.999) and Biojector: 24/25 (CI 0.804, 0.991) was noted, but-22 of 25 patients experienced pain with needle administration: (with VAS = 0; 3/25 [CI: 0.044, 0.302]) (posterior probability [PP] > 0.999). The corresponding VAS scores (means +/- SD) were 0.00 +/- 0.00, 0.04 +/- 0.20, and 2.4 +/- 2.23 (p < 0.001). No pain by proportion of PIS = 0 with MedEJet: 25/25 (CI: 0.868, 0.999 and Biojector: 23/25 (0.749, 0.976) was noted, but pain in 20/25 was felt with the needle: 5/25 (CI: 0.090, 0.394) (PP > 0.999). The corresponding PIS scores were 0.00 +/- 0.00, 0.16 +/- 0.55, and 1.24 +/- 1.00 (p < 0.001). At i.v. catheterization, no pain by proportion of VAS = 0 with MedEJet: 22/25 (CI: 0.698, 0.956) or Biojector: 21/25 (CI: 0.651, 0.934) was noted; but pain in 19/25 with needle administration was experienced: 6/25 (CI: 0.116, 0.436) (PP > 0.999). The corresponding scores were 0.12 +/- 0.33, 0.44 +/- 0.20, and 1.64 +/- 1.50 (p < 0.001). No pain by proportion of PIS = 0 with MedEJet: 24/25 (CI: 0.804, 0.991) or Biojector: 24/25 (CI: 0.804, 0.991) was noted, but pain was apparent in 12/25 with needle administration: 13/25 (CI: 0.334, 0.701) (PP > 0.999). The corresponding scores were 0.00 +/- 0.00, 0.00 +/- 0.00, and 0.76 +/- 0.88 (p < 0.001). Cost per application: MedEJet = $0.13; needle/syringe = $0.50; Biojector = $0.94. CONCLUSIONS: Almost completely painless i.v. catheterization was carried out by jet injection of lidocaine, but needle infiltration produced discomfort or pain and did not significantly reduce discomfort or pain at the i.v. needle insertion. PMID- 10386278 TI - Intraocular pressure changes during rapid sequence induction and intubation: a comparison of rocuronium, atracurium, and succinylcholine. AB - STUDY OBJECTIVE: To compare changes in intraocular pressure (IOP) during rapid sequence induction and intubation following rocuronium, succinylcholine, and atracurium. DESIGN: Open-label, prospective, randomized study. SETTING: Operating room at the Eye Foundation Hospital (University of Alabama at Birmingham) PATIENTS: 45 ASA physical status I, II, and III patients, aged 18 to 65 years, scheduled for elective eye surgery with general anesthesia. INTERVENTIONS: Anesthesia was rapidly induced in unpremedicated patients with a fixed combination of midazolam 0.025 mg/kg, alfentanil 0.025 mg/kg, and propofol 1.5 mg/kg. Intubation was performed, as clinically indicated, approximately 60 seconds following administration of rocuronium 0.6 mg/kg, atracurium 0.5 mg/kg, or succinylcholine 1 to 1.5 mg/kg. MEASUREMENTS AND MAIN RESULTS: Intraocular pressure was measured before induction of anesthesia (baseline), following anesthesia induction and administration of muscle relaxant (before intubation), and after intubation. The percent change in IOP from baseline was significantly decreased in the rocuronium group compared with the succinylcholine group (p = 0.046) before intubation. This trend continued after intubation, but the difference was no longer significant (p = 0.070). Intubation scores for rocuronium and succinylcholine groups were similar, and both scores were superior to that for the atracurium group (p = 0.002). CONCLUSION: Intraocular pressure can be controlled during emergency induction of anesthesia and intubation with adequate depth of anesthesia and muscle relaxation. Rocuronium, succinylcholine, and atracurium all provided sufficient muscle relaxation to achieve successful intubation and no increase in IOP. However, rocuronium 0.6 mg/kg provided significantly better intubating conditions compared with atracurium, and it resulted in a significantly greater decrease in IOP compared with baseline than succinylcholine. PMID- 10386279 TI - Renin-aldosterone system and atrial natriuretic peptide during anesthesia in orthopedic patients over 80 years of age. AB - STUDY OBJECTIVES: To investigate the changes in plasma atrial natriuretic peptide, renin activity, and aldosterone during isoflurane anesthesia in patients over 80 years. DESIGN: Prospective, randomized, controlled study. SETTING: Operating rooms and postanesthesia recovery room of Hirosaki University Hospital and Hakodate Watanabe Hospital. PATIENTS: 36 patients undergoing reduction of femur neck fracture (18 patients ranging in age from 80 to 99 years and 18 patients ranging in age from 40 to 59 years as control). INTERVENTION: In all patients, anesthesia was induced with intravenous (i.v.) thiopental sodium 3 to 5 mg/kg succinylcholine 0.5 to 1.0 mg/kg for facilitating tracheal intubation and was maintained with 1.2% to 2.0% isoflurane in 50% oxygen. MEASUREMENTS AND MAIN RESULTS: Plasma atrial natriuretic peptide (ANP), plasma renin activity (PRA), and plasma aldosterone (PA) levels were assayed. Blood samples were obtained on the following occasions: before the induction of anesthesia, 15 minutes after skin incision, 90 minutes after anesthesia induction, and the 60 minutes after the end of surgery. Plasma renin activity and PA levels in patients from 40 to 59 years increased significantly 90 minutes after induction, whereas PRA and PA levels in patients over 80 years were unchanged. There were significant differences in PRA and PA levels between both groups at any time of measurements. Plasma ANP levels of patients over 80 years were significantly elevated at 90 minutes induction. Plasma ANP levels in patients over 80 years at 90 minutes after the induction and 60 minutes after the end of surgery were significantly higher than those of patients from 40 to 59 years. Plasma renin activity in hypertensive patients over 80 years at 90 minutes after the induction was significantly lower than that observed in normotensive patients. The renal loss of sodium was increased in the hypertensive patients. CONCLUSIONS: Orthopedic patients over 80 years of age have decreased PRA and PA, increased ANP, and renal loss of sodium as compared with patients 40 to 59 years, during isoflurane anesthesia. Plasma renin activity at 90 minutes after induction was decreased in hypertensive patients over 80 years, but PA and ANP were not affected by hypertension during anesthesia. PMID- 10386280 TI - Combination of granisetron and droperidol in the prevention of nausea and vomiting after middle ear surgery. AB - STUDY OBJECTIVES: To evaluate the efficacy and safety of granisetron-droperidol combination for the prevention of postoperative nausea and vomiting (PONV) after middle ear surgery. DESIGN: Prospective, randomized, double-blind study. SETTING: University hospital. PATIENTS: 150 ASA physical status I patients (108 females, 42 males) scheduled for elective middle ear surgery. INTERVENTIONS: Patients received granisetron 40 micrograms/kg (n = 50), droperidol 20 micrograms/kg (n = 50), or granisetron 40 micrograms/kg plus droperidol 20 micrograms/kg (n = 50) intravenously immediately before induction of anesthesia. MEASUREMENTS AND MAIN RESULTS: A standard general anesthetic technique and postoperative analgesia were used throughout the study. A complete response, defined as no PONV and no need for another rescue antiemetic, from 0 to 3 hours after anesthesia occurred in 78%, 56%, and 94% of patients who had received granisetron, droperidol, and granisetron plus droperidol, respectively. The corresponding incidence between 3 and 24 hours after anesthesia was 80%, 52% and 94%. Thus, a complete response within the first 24-hour postanesthetic period was greater in patients receiving granisetron-droperidol combination than in those receiving granisetron alone or droperidol alone (p < 0.05). No clinically adverse events were observed in any of the groups. CONCLUSIONS: A combination of granisetron and droperidol is more effective than droperidol or granisetron alone for the prevention of PONV after middle ear surgery. PMID- 10386281 TI - Delays in the detection of hypoxemia due to site of pulse oximetry probe placement. AB - STUDY OBJECTIVES: To determine if there were any differences in the time to detect hypoxemia related to the site of peripheral pulse oximetry (ear, hand, and foot) during the rapid induction of hypoxemia in healthy volunteers. DESIGN: Repeated-measures, longitudinal, observational study. SETTING: Anesthesia clinical research area of the Department of Anesthesiology. PATIENTS: 13 healthy volunteers, aged 18 to 44 years. INTERVENTIONS: Nellcor N-200 (Nellcor, Inc., Pleasanton, CA) oximeter probes were placed at the ear, hand, and foot. All units were turned on simultaneously with averaging times set for 5 seconds and signals sampled at 2 Hz. A computer-controlled anesthesia circuit was employed to induce mild hypercapnia and hyperoxia (end-tidal gas partial pressures: PETCO2 = 42 +/- 2 mmHg and PETO2 = 130 mmHg) for 5 minutes. PETO2 was then decreased to 45 +/- 2 mmHg over 60 seconds and held at that value for 5 minutes. MEASUREMENTS AND MAIN RESULTS: The mean differences in time (sec) for pulse oximeters to detect hypoxemia (read less than 90%) between probe sites were determined and compared. The following mean differences in time (sec) for pulse oximeters to detect hypoxemia (read less than 90%) between probe sites were found: ear-hand = 6; hand foot = 57; ear-foot = 63. Paired t-tests revealed statistically significant mean time delay differences of 51 seconds (p < 0.005) and 57 seconds (p < 0.005) for ear-hand versus hand-foot and for ear-hand versus ear-foot, respectively. CONCLUSIONS: In healthy volunteers, significant delays in the detection of acute hypoxemia by pulse oximetry occur when pulse oximeters are placed at the toe as compared with probes at either the ear or hand. PMID- 10386282 TI - Treating "rebound" emesis following outpatient gynecologic laparoscopy: the efficacy of a two-dose regimen of droperidol and ondansetron. AB - STUDY OBJECTIVE: To evaluate the efficacy of a two-dose combination of droperidol and ondansetron as compared with single-dose droperidol alone, single-dose combined droperidol and ondansetron, and two-dose droperidol alone, for management of postoperative nausea and vomiting (PONV) among gynecologic laparoscopy outpatients. DESIGN: Randomized, double-blind comparison trial. SETTING: Tertiary outpatient gynecologic unit. PATIENTS: A total of 120 female patients scheduled for gynecologic laparoscopy were enrolled. Patients who had experienced nausea or vomiting, or who had taken drugs with antiemetic action in the 24-hour period prior to the study, as well as breast-feeding mothers, were excluded from participation. INTERVENTIONS: Patients were assigned to four treatment groups: i) single dose of droperidol 1.25 mg, ii) two doses of droperidol 1.25 mg, iii) single dose of droperidol 1.25 mg and ondansetron 4 mg in combination, and iv) two doses of droperidol 1.25 mg and ondansetron 4 mg in combination. The first dose of antiemetic was administered prior to induction and the second dose was given by infusion 4 hours later, prior to discharge. MEASUREMENTS AND MAIN RESULTS: A visual analogue scale (VAS, 10 cm) was used to obtain patients' experience of nausea, vomiting, and pain at 0.5, 1.5, 2.5, and 3.5 hours after arrival at the postanesthetic care unit (PACU). Following discharge, approximately 24 hours after arrival at the PACU, the same measures were obtained by a follow-up interview using a verbal 10-point scale. No significant differences in incidence of PONV were noted among the four treatment groups (p = 0.419). However, both single- and two-dose droperidol and ondansetron combination therapy demonstrated attenuation of PONV severity in the 3.5- to 24 hour postinduction period (p < 0.05). CONCLUSIONS: The findings of this study suggest that prophylactic two-dose combined ondansetron and droperidol offers no added benefit over single-dose therapy for routine use in the gynecologic outpatient population. PMID- 10386283 TI - A reaction to tape after tracheal extubation in a patient with systemic amyloidosis. AB - A reaction to tape after tracheal extubation in a patient with systemic amyloidosis is reported. A patient underwent a right thoracotomy with general anesthesia. A double-lumen tube secured with 1-inch adhesive tape (3M Blenderm, 3M Health Care, St Paul, MN) was used. The same kind of tape was used to cover the eyelids. On removal of the tape, hemorrhagic and purpuric lesions appeared on the skin in regions corresponding to the tape contact area. A diagnosis of amyloidosis was made based on large tongue, lip edema, purpuric and hemorrhagic spots on the skin, congestive heart failure, and skin biopsy. Amyloidosis and its anesthetic implications are discussed. PMID- 10386284 TI - Subdural cannulation and local anesthetic injection as a complication of an intended epidural anesthetic. AB - We report a 52-year-old woman scheduled for laparotomy with combined epidural general anesthesia who experienced abnormal responses to local anesthetic injections administered via the epidural catheter. The catheter subsequently was found to be in the subdural space. A review of the literature is provided. PMID- 10386285 TI - Systolic pressure variation in hemodynamic monitoring after severe blast injury. AB - Fluid management in patients following blast injury is a major challenge. Fluid overload can exacerbate pulmonary dysfunction, whereas suboptimal resuscitation may exacerbate tissue damage. In three patients, we compared three methods of assessing volume status: central venous (CVP) and pulmonary artery occlusion (PAOP) pressures, left ventricular end-diastolic area (LVEDA) as measured by transesophageal echocardiography, and systolic pressure variation (SPV) of arterial blood pressure. All three patients were mechanically ventilated with high airway pressures (positive end-expiratory pressure 13 to 15 cm H2O, pressure control ventilation of 25 to 34 cm H2O, and I:E 2:1). Central venous pressure and PAOP were elevated in two of the patients (CVP 14 and 18 mmHg, PAOP 25 and 17 mmHg), and were within normal limits in the third (CVP 5 mmHg, PAOP 6 mmHg). Transesophageal echocardiography was performed in two patients and suggested a diagnosis of hypovolemia (LVEDA 2.3 and 2.7 cm2, shortening fraction 52% and 40%). Systolic pressure variation was elevated in all three patients (15 mmHg, 15 mmHg, and 20 mmHg), with very prominent dDown (23, 40, and 30 mmHg) and negative dUp components, thus corroborating the diagnosis of hypovolemia. Thus, in patients who are mechanically ventilated with high airway pressures, SPV may be a helpful tool in the diagnosis of hypovolemia. PMID- 10386286 TI - Use of electronic mail for postoperative follow-up after ambulatory surgery. AB - The authors report on a patient who used electronic mail to report satisfactory recovery from ambulatory surgery and anesthesia. The potential benefits and pitfalls of using electronic mail for patient follow-up and communication, as well as research purposes, are reviewed. Potential benefits include cost savings, ease in collecting quality improvement data, and the potential for increased reporting of unpleasant events. Potential pitfalls include lack of universal access (with racial and socioeconomic differentials), privacy and security concerns, and potential slow responses to messages that might require emergent responses or actions. PMID- 10386287 TI - Subcutaneous lidocaine misidentified as cerebrospinal fluid. PMID- 10386288 TI - Back skin movement also causes "walking" epidural catheter. PMID- 10386289 TI - Relationship between systolic pressure variation and left ventricular preload. PMID- 10386291 TI - A "virtual" electronic journal: interactive, innovative, interdisciplinary, and international. PMID- 10386290 TI - New directions in perioperative medicine? PMID- 10386292 TI - Recruitment of house staff into anesthesiology: factors responsible for house staff selecting anesthesiology as a career and individual training program. AB - STUDY OBJECTIVE: To identify factors responsible in the selection of anesthesiology as a career by Mayo Clinic house staff (i.e., residents and clinical fellows); to evaluate their level of satisfaction with their choice of career and training program, and their perceptions of the future for anesthesiology trainees. DESIGN: Cross-sectional analysis using a questionnaire survey of 67 house staff enrolled in the anesthesiology training program during the 1995-1996 academic year. SETTING: Mayo Clinic, Rochester, MN. MEASUREMENTS AND MAIN RESULTS: Forty-eight (72%) of those surveyed responded to the questionnaire. Data were analyzed using the Chi-square and Mann-Whitney rank sum tests. A p-value less than or equal to 0.05 was considered statistically significant. The most frequently cited reasons for selecting anesthesiology as a career included the following: it is a "hands-on" specialty, it involves clinical application of physiology and pharmacology, and it provides immediate gratification in one's work. The most frequently cited reasons for selecting our training program were the diversity of training experience, prestige associated with Mayo Clinic, and employment opportunities following training. Forty-four (92%) felt downsizing of anesthesiology training programs was a national trend, 26 (54%) anticipated difficulty obtaining a job following training, and 16 (33%) felt they had future job security. Overall, 47 (98%) were happy with their career choice, and 40 (83%) would choose anesthesiology as a career if they were now graduating from medical school. All 1996 graduates found suitable employment without difficulty. CONCLUSIONS: Our data indicate that selection of a career in anesthesiology and training program are strongly associated with concerns regarding educational experiences and postgraduate employment opportunities. PMID- 10386293 TI - Continuing medical education and the anesthesiologist. AB - There are a large variety of scheduled activities and courses available to meet the continuing medical education (CME) needs of anesthesiologists. The presentation of CME material varies in format and delivery style. The reasons for attending CME activities include licensure requirements, participation in state and national societies, keeping current with technology, review of old subject material, participation as a lecturer, and other personal reasons. Funding occurs via personal funds, employer support, commercial support, or by research grants. External bodies, such as the American Council of Continuing Medical Education and the American Medical Association, have imposed guidelines in these areas. Methods to evaluate CME activities include retrospective needs analysis based on exit interviews, prospective needs assessment, focus groups, and complex systems such as the CRISIS criteria. Self-directed CME can be evaluated by data collection that identifies how quickly information is received and by the effect of this data on measurable outcome. In the future, CME will increasingly utilize simulators and multimedia computers. Multimedia can bring CME to the physician as opposed to the physician traveling to a CME site. Virtual reality and artificial intelligence are on the horizon and may interface well with the field of anesthesiology due to the technical nature of the discipline and the increasing use of computers and electronic data collection already occurring in clinical practice. PMID- 10386294 TI - Production of probiotic cheese (cheddar-like cheese) using enriched cream fermented by Bifidobacterium infantis. AB - Probiotic cheeses (Cheddar-like cheese) were produced with microfiltered milk standardized with cream enriched with native phosphocaseinate retentate and fermented by Bifidobacterium infantis. During the manufacture and storage of cheeses, viability of the bifidobacteria was determined. Biochemical changes such as proteolysis, sugar metabolism, and organic acids production were estimated. No bifidobacteria growth was observed during cheese-making steps. Bifidobacteria survived very well in cheeses packed in vacuum sealed bags kept at 4 degrees C for 84 d and remained above 3 x 10(6) cfu/g of cheese. No significant difference was observed between cheeses produced with or without bifidobacteria for fat, protein, moisture, salt, ash, or pH. After 12 wk of storage, more than 56% of the as1-CN was hydrolyzed in cheeses that were produced with bifidobacteria and inoculated at 10(8) cfu/g in the cream, and > 45% of hydrolysis was observed in the control cheese. However, no significant differences in the electrophoretic sodium dodecyl sulfate-PAGE patterns were observed in cheeses at any period of storage. At the first day after manufacture, lactose was completely hydrolyzed in cheeses made with bifidobacteria, which suggested high beta-galactosidase activity by B. infantis. Small quantities of acetic acid were detected in bifidus cheeses. The results indicated that B. infantis introduced into hard pressed cheese exhibited excellent viability during storage for 12 wk and could be metabolically active. PMID- 10386295 TI - Sensitivity of Staphylococcus aureus and Lactobacillus helveticus in ovine milk subjected to high hydrostatic pressure. AB - Ovine milk, standardized to 6% fat, was inoculated with Staphylococcus aureus CECT 534 and Lactobacillus helveticus CECT 414 at a concentration of 10(7) cfu/ml and treated by high hydrostatic pressure. Treatments consisted of combinations of pressure (200, 300, 400, 450, and 500 MPa), temperature (2, 10, 25, and 50 degrees C), and time (5, 10, and 15 min). Staphylococcus aureus was highly resistant to pressure; only pressurizations at 50 degrees C of 500 MPa for 15 min achieved reductions of > or = 7.3 log units. For L. helveticus, the number of surviving cells was reduced considerably at pressures of 400 MPa or more (up to 4.5 log units at 50 degrees C for 15 min), and pressure was more effective at low (2 and 10 degrees C) and moderately high (50 degrees C) temperatures than at room temperature (25 degrees C). Both species showed first-order kinetics of destruction in the range 0 to 60 min. The D values for S. aureus were 20 min (2 degrees C at 450 MPa) and 16.7 min (25 degrees C at 450 MPa), and D values for L. helveticus were 7.1 min (2 degrees C at 450 MPa) and 9.1 min (25 degrees C at 450 MPa). Lactobacillus helveticus showed higher rates of survival of pressure than those reported in previous studies for other Lactobacillus spp. PMID- 10386296 TI - Study of the possible mechanisms involved in the mucosal immune system activation by lactic acid bacteria. AB - The induction of a mucosal immune response is not easy due to the development of oral tolerance, but under some conditions, bacteria can activate this immune system. Antigens administered orally can interact with M cells of Peyer's patches or bind to the epithelial cells. We have demonstrated that certain lactic acid bacteria are able to induce specific secretory immunity, and others will enhance the gut inflammatory immune response. The aim of this work was to establish the reason for these different behaviors and to define possible mechanisms involved in the interaction of lactic acid bacteria at the intestinal level. We studied IgA+ and IgM+ B cells comparatively in bronchus and intestine and CD4+ T cells and IgA anti-lactic acid bacteria antibodies in the intestinal fluid, induced by oral administration of Lactobacillus casei, Lb. delbrueckii ssp. bulgaricus, Lb. acidophilus, Lb. plantarum, Lb. rhamnosus, Lactococcus lactis, and Streptococcus salivarius ssp. thermophilus. The increase in the IgA+ B cells in the bronchus means that these lactic acid bacteria were able to induce the IgA cycle by interaction with M cells from Peyer's patches or intestinal epithelial cells. The IgM+ cells increased when the stimulus did not induce the switch from IgM+ to IgA+. The increase in the CD4+ cells suggests interaction of Peyer's patches and enhancement of the B- and T-cell migration. The anti-lactic acid bacteria antibody is related to the processing and presentation of the microorganisms to the immune cells. We demonstrated that Lb. casei and Lb. plantarum were able to interact with Peyer's patch cells and showed an increase in IgA-, CD4+ cells, and antibodies specific for the stimulating strain. Lactobacillus acidophilus induced gut mucosal activation by interaction with the epithelial cells without increase in the immune cells associated with the bronchus. Although Lb. rhamnosus and Strep. salivarius ssp. thermophilus interact with epithelial cells, they also induced an immune response against their epitopes. Lactococcus lactis and Lb. delbrueckii ssp. bulgaricus induced an increase of IgA+ cells entering the IgA cycle but not CD4+ cells; thus, these bacteria would have been bound to epithelial cells that activated B lymphocytes without processing and presenting of their epitopes. We did not determine specific antibodies against Lc. lactis or Lb. bulgaricus. PMID- 10386297 TI - Some practical implications of the milk mucins. AB - Two mucins, large carbohydrate-rich proteins, enter milk from the lactating cell surface by way of the milk fat globule membrane. These mucins relate to a number of practical considerations including physical and flavor properties of milk, mastitis and economically important traits of cattle, and diarrheal disorders of humans and calves (scours). A greater understanding and more effective use of the milk mucins will require additional research. PMID- 10386298 TI - Metabolic responses of lactating dairy cows to 14-day intravenous infusions of glucagon. AB - Twenty cows were assigned at parturition to two groups to study metabolic effects of continuous intravenous infusions of glucagon. Groups were control cows and cows treated with glucagon at 10 mg/d for 14 d starting at d 21 postpartum. Daily blood samples and nine liver biopsies were taken from d 7 to 49 postpartum. Plasma glucagon increased six- to seven-fold during infusions of treated cows. Plasma insulin was increased heterogeneously by glucagon infusions. Plasma glucose increased 11.5 and 9.0 mg/dl during wk 1 and 2 of glucagon infusions. No other plasma metabolites tested (nonesterified fatty acids, beta-hydroxybutyrate, and urea N) were affected by glucagon infusions. Liver glycogen decreased by d 2 of glucagon infusion but was repleted to preinfusion values by d 7 and increased to 169% of the preinfusion baseline values at 3 d after cessation of glucagon. Milk production decreased transiently during glucagon infusions. Both milk production and milk protein percentage decreased during glucagon infusion, which could imply a decreased availability of amino acids for milk protein synthesis. Feed intakes did not increase during glucagon infusions, which was in contrast to the control group. Results indicated that glucagon infusions caused liver glycogenolysis initially and probably enhanced gluconeogenesis but glucagon did not appear to increase lipolysis from adipose tissue in these early lactating dairy cows. PMID- 10386299 TI - Metabolic responses of dairy cows and heifers to various intravenous dosages of glucagon. AB - To evaluate the ability of glucagon to improve carbohydrate status in dairy cows without an increase in blood lipids, glucagon was infused intravenously for 48 h into lactating cows and spayed heifers in three crossover experiments. During Experiment 1, glucagon (5 and 20 mg/d) was infused into four midlactation cows. Experiment 2 involved the infusion of 0, 2.5, 5.0, or 10 mg/d of glucagon into eight heifers; each heifer received two of the dosages. In Experiment 3, four early lactation cows were treated with 5 and 10 mg/d of glucagon. Glucagon consistently increased plasma glucose concentrations in a dose-dependent fashion throughout the 48-h periods. Plasma insulin was increased in a nondose-dependent manner by glucagon in Experiment 1. Plasma urea N was increased when glucagon was administered at 5 mg/d during Experiment 2 and tended to be decreased during Experiment 3. Nonesterified fatty acids in plasma were, in most cases, not affected; however, they were increased by glucagon at 10 mg/d during Experiment 2. Concentrations of beta-hydroxybutyrate were increased only by the 20-mg/d dosage. During Experiment 1, liver glycogen concentrations decreased by 2.1% (wet weight basis) for both dosages of glucagon, and concentrations of total lipid in the liver were increased by 0.6% (wet weight basis) by 20 mg/d of glucagon. Milk fat percentage was increased by glucagon, but milk volume and milk protein production were decreased during Experiment 1. Glucagon improved carbohydrate status over the 48-h periods in all experiments but did not increase plasma nonesterified fatty acids except at the 10-mg/d dosage in Experiment 2. PMID- 10386300 TI - Alleviation of fatty liver in dairy cows with 14-day intravenous infusions of glucagon. AB - Twenty multiparous cows were fed additional concentrate during the final 30 d prepartum to cause susceptibility to fatty liver. From 14 to 42 d postpartum, all cows were subjected to a protocol to induce fatty liver and ketosis. To test glucagon as a treatment for fatty liver, either glucagon at 10 mg/d or excipient was infused via the jugular vein from 21 to 35 d postpartum. All cows had fatty liver at 14 d postpartum and became ketonemic and hypoglycemic during the induction of ketosis. Glucagon increased plasma glucose to 142% of that of controls throughout the 14-d treatment. The hypoinsulinemia present in cows with fatty liver was not affected by glucagon. Plasma beta-hydroxybutyrate and nonesterified fatty acids were decreased by glucagon. At 6 d postpartum, liver triacylglycerol averaged 12.9% of liver (wet weight basis). Glucagon had decreased triacylglycerol content of livers by 71% at d 35. Glycogen was 1.0% of the wet weight of livers at 6 d in milk, but it was decreased by glucagon to 0.5% at 2 d after glucagon began. Glycogen then increased in cows treated with glucagon until at 38 d in milk liver glycogen was 3.7% versus 1.6% in controls. Our results document that glucagon decreases the degree of fatty liver in early lactation dairy cows, which also decreases the incidence of ketosis after alleviation of fatty liver. PMID- 10386301 TI - Regulation of messenger ribonucleic acid expression for gluconeogenic enzymes during glucagon infusions into lactating cows. AB - The effects of glucagon infusions on expression of mRNA for enzymes that regulate gluconeogenesis were studied in lactating cows. Normal cows and cows with fatty liver that were susceptible to ketosis were assigned to either glucagon-treated or control groups. Glucagon at 0 or 10 mg/d was infused for 14 d beginning at d 21 postpartum. In normal cows, glucagon infusions increased concentrations of both plasma glucagon and glucose, which caused plasma insulin to increase. Consequently, hepatic phosphoenolpyruvate carboxykinase mRNA decreased during wk 1 of glucagon infusions. Glucagon infusions into cows with fatty liver also increased plasma glucagon and glucose, but concentrations of plasma insulin and hepatic phosphoenolpyruvate carboxykinase mRNA did not change. More phosphoenolpyruvate carboxykinase mRNA was present in the livers of cows with fatty liver than in livers of normal cows. In a follow-up experiment with midlactation cows, 3.5-h infusions of glucagon at 14 mg/d increased plasma glucose and insulin and decreased plasma nonesterified fatty acids and hepatic glycogen. Hepatic phosphoenolpyruvate carboxykinase mRNA was decreased 41%, pyruvate carboxylase mRNA was increased 50%, but fructose-1,6-bisphosphatase mRNA did not change. We conclude that the expression of the hepatic phosphoenolpyruvate carboxykinase gene in normal cows is inhibited by insulin to balance elevated carbohydrate status during glucagon infusions; however, inhibited expression of hepatic phosphoenolpyruvate carboxykinase mRNA probably is not involved in the pathogenesis of lactation ketosis. PMID- 10386302 TI - Effect of overfeeding during the dry period on the rate of esterification in adipose tissue of dairy cows during the periparturient period. AB - The in vitro rate of esterification of fatty acids in adipose tissue was compared between cows that were fed at restricted energy intake and cows that were overfed during the dry period. Subcutaneous adipose tissue was biopsied at -1, 0.5, 1, and 3 wk from parturition. The basal in vitro rate of esterification was quantified, as well as the rate of esterification after the addition of glucose or glucose plus insulin. The basal rate in adipose tissue from overfed cows at -1 wk was higher than in adipose tissue from cows that were fed at restricted energy intake and indicated enhanced storage of triacylglycerols in adipose tissue of overfed cows at that time. The rate of esterification after the addition of glucose or glucose plus insulin was increased in both groups at each sampling time, but the mean rates, expressed as a percentage of the basal rates, were lower for overfed cows than for cows that were fed at restricted energy intake at 0.5 and 1 wk. Although the addition of glucose or glucose plus insulin increased esterification rates in adipose tissue from both groups of cows, adipose tissue from overfed cows was less sensitive to the addition of these compounds. In conclusion, overfeeding during the dry period predisposed cows to accumulate fat in adipose tissue during the prepartum period. The smaller increase in esterification rate after the addition of glucose or glucose plus insulin in adipose tissue of overfed cows indicates a lower ability of the adipose tissue to esterify circulating fatty acids or to reesterify mobilized fatty acids, which, combined with higher rates of lipolysis postpartum, contributes to continuously elevated concentrations of circulating nonesterified fatty acids postpartum, leading to a more severe hepatic lipidosis in overfed cows. PMID- 10386303 TI - Diagnostic methods for the detection of subacute ruminal acidosis in dairy cows. AB - Two experiments were conducted 1) to validate a field protocol for the determination of ruminal pH and 2) to develop a strategy to interpret ruminal pH data from groups of cows. In the first experiment, ruminal fluid was collected from 30 lactating dairy cows. Ruminal fluid pH was 0.28 pH units lower for fluid collected by rumenocentesis than for fluid collected through a ruminal cannula. Concentrations of volatile fatty acids were correspondingly higher in samples collected by rumenocentesis. A portable pH meter capable of measuring pH of a very small volume of ruminal fluid yielded very similar pH readings as did a standard meter with a pH probe. Filtration or aspiration of ruminal fluid had no effect on pH. In the second experiment, a strategy was developed to use ruminal pH values from a subsample of cows to distinguish between groups fed either a low or higher forage diet. Groups could be distinguished using a cut point of 5.5 ruminal pH, a sample size of 12 cows, and a critical value of 3 or more cows below the cut point. This strategy had the lowest theoretical error rate for herds with either a high or low prevalence of cows with a low ruminal pH. PMID- 10386304 TI - A noninvasive radiotelemetry system to monitor heart rate for assessing stress responses of bovines. AB - A noninvasive radiotelemetry system was developed to monitor heart rates of cows and to view and analyze data. The system was validated by comparing heart rate data of two restrained heifers collected simultaneously using telemetric and direct electrocardiogram measurements and by acquiring data over 72 h from two dry cows housed in an experimental handling facility consisting of a free-stall pen, a holding pen, a pass-through stall, and a second holding pen. Telemetric and direct measurements in response to pharmacological elevation of heart rates were essentially identical. For cows in the experimental facility, peristimulus time histograms indexed to standing or lying showed that average heart rates for cows increased 4.0 +/- 1.4 beats/min after cows stood and decreased 4.8 +/- 1.0 beats/min after cows lay. Similarly, the average heart rate for the cow naive to the facility increased from 60 to 86 beats/min and remained elevated for 6.3 min when heart rate was indexed to maximal heart rate within +/- 3 min of entry into the pass-through stall. Heart rate for the naive cow increased consistently from around 60 to over 160 beats/min during repeated agonistic encounters between animals. Heart rate for the other cow was not affected by the encounters. These results show clearly that heart rate can be used to monitor animal anxiety. PMID- 10386305 TI - Effects of diseases on test day milk yield and body weight of dairy cows from Danish research herds. AB - The pre- and postdisease interrelationships of energy corrected test day milk yield and body weight of dairy cows caused by mastitis, three reproductive disorders (retained placenta, metritis, cystic ovaries), and seven metabolic disorders (milk fever, ketosis, decreased rumen motility, enteritis, left displaced abomasum, right displaced abomasum, and off feed) were quantified by using mixed models analysis with repeated measures of continuous data. The data were weekly recordings from 4414 lactations collected in three Danish research herds. High milk yield was a risk factor for ketosis and enteritis. Heavier primiparous cows were more likely to contract mastitis. Milk yield was decreased for a disease-specific period for all study diseases except cystic ovaries and right displaced abomasum. Metabolic disorders had a detrimental effect on body weight. The highest weight loss (69 kg) was associated with left displaced abomasum. The persistence of the weight loss differed considerably among study diseases. Almost all weight loss occurred up to and including the initial week after diagnosis, which emphasized the detrimental effect of the subclinical stage. However, weekly measured body weight seemed superior to weekly energy corrected test day milk yield for disease detection only for decreased rumen motility and left displaced abomasum. This study demonstrates the importance of the predisease level for accurate estimation of the loss of milk yield and body weight from disease. PMID- 10386306 TI - Intracellular accumulation, subcellular distribution, and efflux of tilmicosin in bovine mammary, blood, and lung cells. AB - Tilmicosin is a semisynthetic macrolide antibiotic currently approved for veterinary use in cattle and swine to combat respiratory disease. Because the concentrations of tilmicosin are generally low in bovine serum, the interaction of tilmicosin with three types of bovine phagocytes (monocyte-macrophages, macrophages, and neutrophils from blood, lungs, and mammary gland, respectively) and mammary gland epithelial cells was evaluated to provide an understanding of potential clinical efficacy. After incubation with radiolabeled tilmicosin, uptake was determined and expressed as the ratio of the intracellular to the extracellular drug concentration. Accumulation of tilmicosin at 4 h of incubation by the alveolar macrophages (Cc/Ce 193) was 4 to 13 times more than that observed in monocyte-macrophages (Cc/Ce 43), neutrophils, (Cc/Ce 13), or mammary epithelial cells (Cc/Ce 20). Subcellular distribution showed that 70 to 80% of tilmicosin was localized in the lysosomes. Uptake in mammary gland cells was dependent on cell viability, temperature, and pH, but was not influenced by metabolic inhibitors or anaerobiosis. However, lipopolysaccharide exposure increased tilmicosin uptake by the bovine mammary macrophages and epithelial cells. When neutrophils and epithelial cells were incubated in the presence of tilmicosin and extracellular tilmicosin was then removed, 40% of the intracellular tilmicosin remained cell associated after 4 h of incubation (i.e., 60% effluxed), but only 25% remained in macrophages. These in vitro interactions of tilmicosin with bovine phagocytes and epithelial cells suggest an integral role in effecting clinical efficacy. PMID- 10386307 TI - Effects of clinical mastitis on milk yield in dairy cows. AB - The effect of clinical mastitis on milk yield was studied in 24,276 Finnish Ayrshire cows that calved in 1993 and were followed for one lactation (i.e., until culling or the next calving). Cows that had only mastitis, but no other diseases, and cows that had no diseases (healthy cows) during the lactation were included in the study. Monthly test day milk yields were treated as repeated measurements within an animal in a mixed model analysis. Mastitis index categories were created to relate the timing of mastitis to the test day milk measures. Statistical models (a separate model for each parity) included fixed effects of calving season, stage of lactation, and mastitis index. An autoregressive correlation structure was used to model the association among the repeated measurements. The effect of mastitis occurring at different periods during the lactation was studied. The daily loss during the first 2 wk after the occurrence of mastitis varied from 1.0 to 2.5 kg, and the total loss over the entire lactation varied from 110 to 552 kg and depended on parity and the time of mastitis occurrence. Regardless of the time of occurrence during the lactation, mastitis had a long-lasting effect on milk yield; cows with mastitis did not reach their premastitis milk yields during the remainder of the lactation after onset of the disease. PMID- 10386308 TI - Determinants of success or failure in the elimination of major mastitis pathogens in selective dry cow therapy. AB - Discriminant factors for the elimination of major pathogens (mainly Staphylococcus aureus and Streptococcus dysgalactiae) were identified from a randomized, double-blind, field study on dry cow therapy. From an original study of 686 cows, 269 fulfilled the criteria for this analysis: 93 from the control group, 83 from group C (treated with Benestermycin vet. "LEO") and 93 from group D (treated with Leocillin with Dihydrostreptomycin vet. "LEO"). A "success" cow was defined as a cow that had all quarters healthy in two tests after dry period. Isolation of a major pathogen (mainly Staph. aureus or Strep. dysgalactiae) in any quarter in both samples after the dry period was defined as "failure." Better elimination was associated with short-acting preparations (therapy D) rather than with long-acting therapy C (odds ratio = 0.32), as was a smaller mean value of the last three composite milk somatic cell counts (CMSCC) before dry cow therapy. Cows with a major pathogen in the right hind quarter at drying off or at least one case of acute clinical mastitis during the previous lactation were more likely to have a major pathogen in the next lactation (odds ratio = 4.1 and 3.6, respectively). Young cows without major pathogens at drying off should not be recommended for dry cow therapy, and cows with high CMSCC should be considered for culling if their profiles also show previous acute clinical mastitis in combination with generally high CMSCC and low cure rates in the herd. PMID- 10386309 TI - Effect of milking frequency and pasture intake on milk yield and composition of late lactation cows. AB - Twenty-four monozygous twinsets in late lactation (> 210 d in milk) were used to examine the effects of feed restriction and milking frequency prior to drying off on milk yield and composition in a pastoral dairying system. Cows were assigned to one of four treatment groups for 26 d and were milked either twice or once daily and given either unrestricted or restricted access to feed. Dry matter intakes averaged 16 or 8 kg per cow per day, and diets comprised ryegrass and white clover pasture supplemented with 15% pasture silage. Feed restriction and once daily milking reduced milk yield and increased concentrations of milk fat and protein. Somatic cell count was increased by feed restriction only. Production losses caused by feed restriction were nearly threefold higher than were those for once daily milking. Yields of components that were mammary synthesized and serum derived were reduced by feed restriction, in accordance with milk volume reduction. Plasma lactose concentration increased with once daily milking only and indicated enhanced permeability of mammary tight junctions. Both feed restriction and once daily milking compromised milk quality, but increased leakage of serum components into milk via mammary tight junctions was deemed to occur only for once daily milking. PMID- 10386310 TI - Long-term effects of feeding gossypol and vitamin E to dairy calves. AB - Male Holstein calves were used to test the effect of feeding 400 mg of free gossypol/kg of diet and to determine whether vitamin E could counteract gossypol toxicity. Fifty-two calves were allotted to treatments as follows: 1) soybean meal-based starter; 2) cottonseed meal-based starter; 3) cottonseed meal-based starter + 2000 IU of vitamin E/d per calf, and 4) cottonseed meal-based starter + 4000 IU of vitamin E/d per calf. Vitamin E supplementation (treatments 3 and 4) improved weight gain and feed intake over calves on treatment 1. Gossypol concentrations in plasma were higher in calves on treatments 2, 3, and 4 than in calves on treatment 1; however, no differences were observed among animals receiving the three cottonseed meal diets. Hemoglobin and hematocrit were decreased in calves receiving treatment 2, and vitamin E supplementation counteracted this effect (treatments 3 and 4). Plasma alpha-tocopherol concentrations were not affected by gossypol intake and followed the vitamin E supplementation pattern During the experimental period, 10 calves died, six from treatment 2 and two each from treatments 3 and 4. Necropsy findings from 4 of 10 calves were suggestive of gossypol toxicity. Histopathological examination revealed centrilobular necrosis in the liver and atrophy and vacuolation of cardiocytes. Feeding cottonseed meal caused death of some calves with gossypol related toxicity signs, but did not decrease plasma alpha-tocopherol; however, vitamin E supplementation increased performance and may have conferred some protection against gossypol toxicity. PMID- 10386311 TI - Effects of extrusion of grain and feeding frequency on rumen fermentation, nutrient digestibility, and milk yield and composition in dairy cows. AB - The effect of corn extrusion and feeding frequency on ruminal and postruminal digestibility and milk yield was studied in cows fed a high concentrate diet. Four Israeli Holstein cows fitted with rumen and abomasal cannulas were used. The experiment was arranged as a 2 x 2 factorial design, with two diets and two feeding frequencies (two or four meals per day). One diet contained 40% ground corn. In the second diet, half of the ground corn was replaced with extruded corn. Feeding cows the extruded versus ground corn diet decreased ruminal ammonia N and plasma urea N concentrations, increased postruminal digestibility of nonstructural carbohydrates, reduced dry matter intake, decreased yield of milk and milk components, and increased efficiency of milk energy and milk protein synthesis. The inclusion of extruded corn in the diet did not affect ruminal volatile fatty acid. Increasing the feeding frequency reduced the diurnal variation in ruminal pH, ruminal ammonia, and plasma urea, and increased dry matter intake--considerably more in the cows fed ground versus extruded corn--and improved postruminal organic matter, nonstructural carbohydrate, and crude protein digestibility. Total tract digestibility of organic matter and crude protein and milk yield and composition were also increased when cows were fed four versus two meals. Concurrent with the feeding frequency and grain processing effect, an increase in rumen-undegradable protein flow was related to increased digestion of nonstructural carbohydrate postruminally (r = 0.54). We concluded that for cows fed high-starch diets more frequent meals are useful for improving postruminal digestibility and milk yield and composition. PMID- 10386312 TI - Milk urea nitrogen target concentrations for lactating dairy cows fed according to National Research Council recommendations. AB - The objectives of this study were to develop and evaluate a mathematical model to predict milk urea N and to use this model to establish target concentrations. A mechanistic model to predict milk urea N was developed using raw data from 3 studies (10 diets, 40 cows, and 70 observations) and was evaluated with 18 independent studies (89 treatment means). For the independent literature data set, the model prediction error was approximately 35%; the majority of the error was due to variation among experiments. A mean of at least 25 cows was determined to be necessary for reliable model predictions. This model, which uses such data as protein intake and milk production, was used to predict milk urea N concentrations when cattle are fed according to National Research Council recommendations. Target values calculated in this manner for a typical lactation were 10 to 16 mg/dl, depending on days in milk. Target concentrations were sensitive to changes in milk production and amount of N intake and were relatively insensitive to body weight, parity, and grouping strategy. Analysis of data from the Lancaster Dairy Herd Improvement Association (n = 133,057) indicated that cows in the region were being fed diets containing approximately 17% crude protein, regardless of parity. A comparison to target milk urea N concentrations for this data indicated that cows were being fed 8 to 16% more protein than recommended by the National Research Council. Target milk urea N concentrations have been established, and dairy farmers now have a definitive way to interpret milk urea N concentrations. PMID- 10386313 TI - Glucose kinetic responses to protein supplementation and exogenous somatotropin in late gestation dairy cows. AB - Glucose kinetics were measured in late gestation multiparous Holstein dairy cows fed diets with different amounts of dietary crude protein (13.3 vs. 17.8%), with and without exogenous somatotropin. The trial was conducted as a completely randomized design; 35 cows were used in the final analysis. Kinetic measurements were made using the single injection technique with uniformly labeled 13C-labeled glucose. A diet that contained 17.8% crude protein appeared to increase glucose utilization without a corresponding increase in supply. The evidence was an increased rate of glucose disposal. In contrast, exogenous bovine somatotropin (Posilac, Monsanto Co., St. Louis, MO) appeared to enhance glucose conservation, as indicated by a tendency for a decreased fractional catabolic rate and an increased glucose pool size. Somatotropin appeared to modify glucose metabolism in a pattern favorable for supporting terminal fetal development and lactogenesis and for maintaining or enhancing maternal glycemia. The latter pattern could have implications for improving the health of periparturient dairy cows. PMID- 10386314 TI - Effect of energy and protein density of prepartum diets on fat and protein metabolism of dairy cattle in the periparturient period. AB - To determine if increased nutrient density in prepartum diets improves nutrient balance of peripartum cows, we blocked 40 Holstein cows and 40 heifers by expected date of parturition and assigned them randomly within blocks to one of four treatment diets varying in density of net energy for lactation (NEL) and crude protein (CP). Diets were 1.30 Mcal of NEL/kg and 12.2% CP, 1.49 Mcal of NEL/kg and 14.2% CP, 1.61 Mcal of NEL/kg and 15.9% CP, and 1.48 Mcal of NEL/kg and 16.2% CP. These diets were fed ad libitum from 25 d prepartum until parturition, and all cows were fed the same diet after calving. Increased nutrient-density of prepartum diets did not decrease feed intake. Compared to animals fed the lowest density, those fed the highest density consumed more NEL (20 vs. 14 Mcal/d) and gained more body condition, backfat, and body weight. They also had less nonesterified fatty acids in plasma (176 vs. 233 microM) and more insulin-like growth factor-I in plasma (472 vs. 390 ng/ml) during the last 2 wk prepartum and less triglyceride in liver at parturition (0.9 vs. 1.5%, wet tissue basis). Quadratic effects of energy density were not observed, and the addition of protein in the medium energy diet had no effect. Prepartum diets did not alter any variables during lactation. In conclusion, increasing the energy and protein density up to 1.6 Mcal of NEL/kg and 16% CP in diets during the last month before parturition improves nutrient balance of cattle prepartum and decreases hepatic lipid content at parturition. PMID- 10386315 TI - Effect of coating whole cottonseed on performance of lactating dairy cows. AB - Thirty-six lactating Jersey cows were used in a randomized block design to determine the effect of coating whole fuzzy cottonseed to improve handling characteristics on intake, milk yield, apparent digestibility of nutrients, and blood gossypol concentrations. Treatments included whole cottonseed at 15% of dietary dry matter either as whole cottonseed, whole cottonseed coated with 5% gelatinized corn starch, or whole cottonseed coated with 5% corn starch plus 10% maltodextrin sugar. Dry matter intake, milk yield, percentage of milk protein and lactose, and yield of milk components were not different among treatments; however, the percentage of milk fat was depressed when maltodextrin sugar was included in the coating. When in vitro fermentations of mixed ruminal microorganism were conducted, final pH was lower and concentrations of total fatty acids, propionate, and L-lactate were higher for whole cottonseed coated with starch and sugar compared with uncoated cottonseed. Nutrient intake was similar among treatments, but the apparent digestibility of acid and neutral detergent fiber was reduced when coated cottonseed were fed. Total plasma gossypol concentration was higher for the cottonseed coated with starch compared with cottonseed coated with starch and sugar, but the difference was not of biological significance. Results of this study indicate that coating whole cottonseed with starch does not alter its palatability or nutrient value for supporting milk yield, but a reduction in fiber digestibility was observed. Inclusion of 10% maltodextrin sugar in the coating altered ruminal fermentation and resulted in a depressed percentage of milk fat. PMID- 10386316 TI - Performance of lactating dairy cows fed whole cottonseed coated with gelatinized cornstarch. AB - The handling characteristics of whole cottonseed are improved by coating with gelatinized cornstarch, but limited information is available on the effects of feeding the coated cottonseed to lactating dairy cows. Thirty-six lactating Jersey cows were used in a crossover design trial with 4-wk experimental periods to evaluate the influence of coating whole cottonseed with 2.5% gelatinized cornstarch on dry matter intake, milk yield, and composition. Cows were fed diets containing 10.2% alfalfa-orchardgrass hay, 45.2% corn silage, 15.0% coated or uncoated whole cottonseed, and 29.6% concentrate for ad libitum consumption. Coating whole cottonseed with gelatinized cornstarch tended to reduce dry matter intake, which averaged 16.2 and 15.9 kg/d for uncoated and coated cottonseed, respectively. Milk yield and composition were similar for uncoated and coated cottonseed. The yield of energy-corrected milk per unit of dry matter consumed was greater with coated cottonseed. Cows fed coated cottonseed gained body weight, but cows fed uncoated cottonseed lost weight. Concentrations of plasma urea were similar among treatments; however, NEFA concentrations were lower for cows fed coated whole cottonseed. Results of this trial indicate that coating whole cottonseed with 2.5% gelatinized cornstarch does not alter its feeding value for lactating dairy cows. PMID- 10386317 TI - The influence of acidic diets on the acid-base balance of dry cows and the effect of fertilization on the mineral content of grass. AB - To investigate the safety and practicality of an acidic concentrate in milk fever prevention, the pH, carbon dioxide, standard bicarbonate, and base excess of whole blood and the pH in the urine were measured in three treatment groups of dry cows after 14 and 21 d of feeding an acidogenic diet (experiment 1). The dietary cation-anion differences (DCAD) of cows on treatments 1 (n = 11), 2 (n = 13), and 3 (n = 12) were +2275, -262, and -1185 meq/d, respectively. No changes in any parameters were found from the beginning to the end of the experiment in cows on treatment 1. In cows on treatment 2, a significant reduction in urine pH was observed, and in cows on treatment 3 significant decreases in all parameters except blood pH were observed. Mineral analyses of grass samples from fields fertilized with N from NH4NO3, Ca(NO3)2, or (NH4)2SO4, with S from (NH4)2SO4, and with different amounts of K from KCl or K2SO4 and of Cl from KCl revealed DCAD ranging from -14 to +726 meq/kg of dry matter (experiment 2). Fertilization with Cl increased the chloride concentration in the crop and had the largest effect on DCAD. The results indicate that the use of acidic concentrates is not a health hazard for dry cows, at least not when the DCAD is greater than about -1200 meq/d or about -140 meq/kg of dry matter. PMID- 10386319 TI - Clinical mastitis in Norwegian cattle: frequency, variance components, and genetic correlation with protein yield. AB - Records of clinical mastitis in first lactation Norwegian Cattle from 1978 onward were analyzed. Variance components for clinical mastitis were estimated with a linear sire model using records of more than 1.2 million cows from 2043 sires, resulting in heritability estimates of 0.035. Different strategies for extracting data gave very similar results, and estimated heritability for mastitis was the same with univariate and bivariate (with protein yield) analyses, which indicates that selection bias caused by correlated responses from other traits in the breeding goal is not a problem with this data set. The estimated genetic correlation between clinical mastitis and protein yield is 0.25. PMID- 10386318 TI - Hypocalcemia induced by intravenous administration of disodium ethylenediaminotetraacetate and its effects on excretion of calcium in urine of cows fed a high chloride diet. AB - Evidence supports the theory that a diet that is rich in nonmetabolizable anions fed to dairy cows during the dry period reduces the risk of hypocalcemic paresis puerperalis. When cows are fed a diet that is rich in anions instead of cations, more Ca is absorbed in the intestine and excreted in urine. We hypothesized that, in cows fed a diet that was rich in anions, the increased flow of Ca through the body could be drained to support the maintenance of plasma Ca concentration around parturition. The hypothesis was tested by binding plasma Ca through intravenous administration of Na2-EDTA and measuring excretion of Ca in urine. In a 2-period x 14-d crossover study, six, nonpregnant, nonlactating, multiparous cows were fed either a diet that was rich in cations (dietary cation-anion difference = +332 meq/kg of dry matter) or rich in anions (dietary cation-anion difference = -230 meq/kg of dry matter). On the last day of each feeding period, Na2-EDTA was infused intravenously until the amount of plasma Ca that was not bound to EDTA reached approximately 1 mmol/L. The amount of EDTA that could be infused was significantly greater when the cows were fed the diet that was rich in anions. During the infusion of Na2-EDTA the rate of Ca excretion in urine dropped to almost 0 when the diet that was rich in anions was fed. After feeding the diet that was rich in cations, excretion of Ca in urine was negligible and was not reduced further by Na2-EDTA infusion. Thus, in cows fed a diet that was rich in anions, the Ca intended for excretion with urine can be used when plasma Ca is under stress as would occur at the onset of lactation. However, the amount of Ca derived from plasma, interstitial fluid, and the skeleton during Na2-EDTA infusion was quantitatively much more important to the supply of Ca than was the reduction in excretion of Ca in urine. Most likely, this relationship would also be true when the production of colostrum begins. PMID- 10386320 TI - Genetic correlations among protein yield, productive life, and type traits from the United States and diseases other than mastitis from Denmark and Sweden. AB - Sire genetic evaluations for protein yield, productive life, and selected type traits from the US were correlated with sire evaluations for disease from Denmark and Sweden and were then adjusted to approximate genetic correlations. Disease categories from Denmark included reproductive diseases, foot and leg diseases, metabolic and digestive diseases, and all diseases other than mastitis. Genetic evaluations for Denmark were from separate analyses for each disease category using a multiple-trait sire model with first, second, and third lactations handled as multiple traits. Evaluations from Sweden for all diseases other than mastitis were from a single-trait sire model using only first lactations. In addition, Danish and Swedish genetic evaluations were regressed on US type evaluations to test for quadratic relationships. Relationships were based on 104 bulls with US and Danish evaluations (88 with US type) and 84 bulls with US and Swedish evaluations (83 with US type). Genetic correlations between US protein yield and diseases were unfavorable, but correlations were favorable between productive life and disease. Genetic correlations among US type and diseases were around zero, except for correlations with US dairy form (range -0.34 to -0.73). Genetic correlations calculated from residual correlations (adjusted for predicted transmitting abilities for milk) between productive life and diseases were favorable (range 0.29 to 0.51). Genetic correlations calculated from residual correlations (adjusted for predicted transmitting abilities for milk) between dairy form and diseases ranged from -0.10 to -0.53. Selection for increased productive life may reduce disease occurrences, but selection for higher dairy form scores will increase disease occurrences. PMID- 10386321 TI - Conjugated linoleic acid and other anticarcinogenic agents of bovine milk fat. AB - Prevention is an important strategy for conquering cancer. Milk fat contains a number of components, such as conjugated linoleic acid, sphingomyelin, butyric acid, ether lipids, beta-carotene, and vitamins A and D that have anticancer potential. Conjugated linoleic acid inhibits the growth of a number of human cancer cell lines and suppresses chemically-induced tumor development at a number of sites in animal models. As little as 0.1% of dietary conjugated linoleic acid inhibits the development of rat mammary tumors, independent of the amount and type of fat in the diet. Sphingomyelin, through its metabolites ceramide and sphingosine, participates in multiple antiproliferative pathways associated with suppression of carcinogenesis. Dietary sphingomyelin inhibits murine colon tumor development. Butyric acid, uniquely present in ruminant milk, is a potent antineoplastic agent and may ameliorate its potency through synergy with other milk fat components. Dietary butyric acid inhibits mammary carcinoma development in rats. In humans, ether lipids, beta-carotene, and vitamins A and D are associated with anticancer effects. Cows have the ability to extract anticarcinogenic components from pasture and feed and transfer them to milk. Use of genetic engineering and other techniques to increase the range and level of anticarcinogens in pasture and supplements may increase the anticancer potential of milk. PMID- 10386322 TI - Animal model genetic evaluation of type traits for five dairy cattle breeds. PMID- 10386323 TI - Motivation, mobilization, and monitoring: the role of groups in health policy. PMID- 10386324 TI - Social movements as catalysts for policy change: the case of smoking and guns. AB - Social movements organized around perceived threats to health play an important role in American life as advocates for change in health policies and health behaviors. This article employs a framework drawn from social movement and related sociological theories to compare two such movements: the smoking/tobacco control movement and the gun control movement. A major purpose of the article is to identify specific social movement ideologies and actions that are more or less likely to facilitate achievement of the movement's health policy objectives. The article concludes that the success of health-related social movements is associated with (1) the articulation of a socially (as well as scientifically) credible threat to the public's health, (2) the ability to mobilize a diverse organizational constituency, and (3) the convergence of political opportunities with target vulnerabilities. PMID- 10386325 TI - The missing millions: organized labor, business, and the defeat of Clinton's Health Security Act. AB - During the battle over comprehensive health care reform in the early 1990s, organized labor was not only unable to put together a winning coalition but also found itself divided and on the defensive as it struggled to prevent any further erosion of the private-sector safety net of the U.S. welfare state. Labor's relative ineffectiveness has deep institutional and political roots and was not merely a consequence of its dwindling membership base. Several key institutions of the private welfare state, notably the Taft-Hartley health and welfare funds and the Employment Retirement Income Security Act (ERISA) preemption, brought the interests of organized labor more closely in line with those of large employers and commercial insurers and aggravated divisions within organized labor and between unions and public interest groups. In addition, several political factors conspired to reinforce labor's tendency to stick to a policy path on health care issues that was predicated on an employer-mandate solution and that had been charted primarily by business and leading Democrats. As a result, organized labor did not emerge from the 1993-1994 struggle with its political base fortified nor with a viable long-term political strategy to achieve universal health care and to shift the political debate over health policy in a more desirable direction. PMID- 10386326 TI - The new NIH and FDA medical research policies: targeting gender, promoting justice. AB - The National Institutes of Health (NIH) and Food and Drug Administration (FDA) have both recently revised their policies regarding the inclusion of women in clinical trials. Pressured by women's health activists and members of Congress, the NIH has vastly improved its policies; it now requires that women and minorities the included in clinical trials and that an analysis of gender and racial differences be performed. The FDA policy states that women and men should be included in clinical trials if both would receive the drug when marketed and that it expects a gender analysis to be performed. The FDA also lifted its 1977 ban on including women of childbearing potential in the early phases of drug studies. Analyzing these NIH and FDA policies according to a gender justice framework, I find that the NIH has moved significantly toward the institution of gender justice as it applies to medical research policies and that the FDA has taken only small steps toward this goal and lags behind the NIH. PMID- 10386327 TI - The implementation and enforcement of tobacco control laws: policy implications for activists and the industry. AB - We examine the process by which antitobacco laws and ordinances were implemented and enforced in seven states and nineteen localities. Our findings indicate that state- and local-level clean indoor air laws were rarely enforced by governmental agencies. Instead, these laws were largely self-enforcing in that changed social norms regarding appropriate smoking behavior led to generally high compliance rates. In contrast, teen access laws were not self-enforcing, but were often enforced through periodic vendor compliance checks. We also found that antitobacco forces did not devote a significant amount of attention of implementation and enforcement issues. Their focus was primarily on enacting new legislation and fighting tobacco industry attempts to weaken existing laws. Our results do not augur well for public health measures that require state-level enforcement and that are opposed by powerful and politically well-connected interests. For tobacco control laws to be effective, public health advocates need to consider the locus of enforcement responsibility and the sanctions available to the enforcement agency, such as license removal by local authorities. These results suggest that failure to specify such mechanisms in the legislation will lead to delays in implementing and enforcing the laws as well as to a number of compliance problems. Antitobacco coalitions will also need to become more actively involved in the implementation and enforcement process. PMID- 10386328 TI - Regulating managed care: interest group competition for control and behavioral health care. AB - In this essay I identify how historic patterns of competition among health care interest groups have simultaneously retained their past contours and also changed significantly as a result of the jolt created by the rise of managed care. I explain why it is that I and other executives of for-profit managed behavioral health care organizations, traditionally advocates of market-based approaches to accountability, have begun to support some versions of regulation by the national government in order to restore consumer confidence in the credibility of managed health plans. Key concepts that are addressed include notions of public accountability, control of the purse strings, consumer protections, and provider privileges. PMID- 10386329 TI - The International Conference on Harmonization Good Clinical Practice guideline. AB - The purposes of the International Conference on Harmonization (ICH) Good Clinical Practice (GCP) Guideline are to protect the rights of human subjects participating in clinical trials and to ensure the scientific validity and credibility of the data collected in human clinical studies. The guiding principle in the guideline is that the rights, safety, and well-being of the trial subject are the most important considerations and should prevail over the interests of science and society. The guideline will have an important and beneficial impact on the clinical trials conducted in the three participating regions (the United States, Europe, and Japan) as well as many other regions throughout the world. In the years to come, it should fulfill its intended purpose of providing for a more economical use of human, animal, and material resources and the elimination of unnecessary delays in the global development and availability of new medicines, and at the same time maintaining safeguards on quality, safety, and efficacy and regulatory obligations to protect public health. PMID- 10386330 TI - Quality assurance reviews: how they differ from peer reviews. AB - Research papers and reports written by scientists and engineers in the United States Environmental Protection Agency are reviewed by the agency's quality assurance staff. EPA papers and reports are subjected to peer reviews that check for the validity of conclusions and the general agreement with the body of technical knowledge in the subject area. Quality assurance reviews differ from peer reviews in that the focus of the quality assurance review is on the following criteria: Consistency: Were reasonable and consistent units of measurement and generally acceptable formulas used throughout? Are the appropriate number of significant figures reported? Correctness: Were matrix compatible methods used? Were measurements within the working range of the method? Can measurements be traced to a recognized standard or source (e.g., the National Institute of Standards and Technology)? Can calculations be verified, starting from representative raw data and proceeding to the summary data presented in the paper or report? Coherence: Do the stated conclusions follow from the data presented? Are the assumptions clearly stated? Are inconsistencies between data and conclusions discussed? Clarity: Are special terms and acronyms defined? Can a person with a general technical background in the subject understand the paper or report? Conformance: Did the study follow the test/quality assurance plan, with appropriate calibrations and other quality control checks, audits, and data validations? If not, is there a discussion of problems? Concordance: Were data quality objectives met? Were the data quality indicator goals achieved for precision, accuracy, representativeness, comparability, and completeness? The importance of these quality assurance review criteria are discussed along with examples from current work. PMID- 10386331 TI - Improving the process quality using statistical design of experiments: a case study. AB - A technique known as Statistical design of experiments is a powerful technique for process characterization, optimization, and modeling. It has been widely accepted in manufacturing industry for improving product performance and reliability, process capability, and yield. This article illustrates the application of statistical design of experiments based on the Taguchi approach in a certain company that manufactures electromagnetic clutch coils. The objective of the study was to improve the quality of the existing process and thereby achieve heightened customer satisfaction for the product. An eight-trial experiment was conducted with the aim of reducing the number of rejects from the process. The expected savings per month was estimated to be over $11,500. The results of the study have provided a greater stimulus for the wider application of statistical design of experiments in other core processes within the company. PMID- 10386332 TI - A general guide for conducting in-process inspections. AB - Quality-assurance inspections are necessary to assure that studies are conducted according to protocol, standard operating procedures, (SOPs) and government regulations. This article describes observation methods used when conducting inspections, the basic steps of an inspection, and the seven inspection principles (protocol, SOPs, reagents/drugs/test article, equipment, personnel, facility, and management) used to complement and standardize an in-process inspection. In addition, differences among preclinical, clinical, and analytical study inspections are discussed and helpful tips are identified. PMID- 10386333 TI - The costs and benefits of getting the ISO 9000 certification in the manufacturing sector in Saudi Arabia. AB - Many Saudi companies, in their journey to improve quality, efficiency and competitiveness, are pursuing and obtaining the ISO 9000 certificate. Many studies have evaluated how to implement ISO 9000 in different sectors, but none have analyzed the effectiveness of ISO 9000 certification (costs and benefits) on improving the overall quality and on meeting expectations. This study addressed these issues by investigating manufacturing organizations in Saudi Arabia that have the ISO 9000 certification. A survey questionnaire was distributed to firms throughout the kingdom. Thirty-two firms participated in the study. Results indicate that increased consistency of operations, improved service, and product quality are among the top motivators for pursuing the ISO certificate. The benefits most often experienced were improved awareness of procedural problems, better management control, keeping existing customers, increased customer satisfaction, and improved customer service. Difficulties experienced during the certification process involved time and cost, but these were not considered to be major problems. A high volume of paperwork was the main problem experienced following initial certification. Respondents in general said that the ISO 9000 certification met their expectations and that their level of satisfaction regarding the impact of ISO 9000 was high. Most recommended that other organizations pursue the certificate. PMID- 10386334 TI - Time to consider the concept of a commensal virus? PMID- 10386335 TI - Specific cleavage of simian virus 40 DNA by restriction endonuclease of Hemophilus influenzae. 1971. PMID- 10386336 TI - Paramyxovirus replication and pathogenesis. Reverse genetics transforms understanding. AB - A recent breakthrough in the field of nonsegmented negative strand RNA viruses (Mononegavirales), including paramyxoviruses, is the establishment of a system to recover an infectious virus entirely from complementary DNA and hence allow reverse genetics. Mutations can now be introduced into viral genomes at will and the resulting phenotypes studied as long as the introduced mutations are not lethal. This technology is being successfully applied to answer outstanding questions regarding the roles of viral components in replication and their contribution to pathogenicity, which are difficult to address using conventional virology. For instance, how the paramyxovirus accessory proteins V and C contribute to actual viral replication and pathogenesis has remained unanswered since their first description more than 20 years ago. Using Sendai virus, which causes fatal pneumonia in mice, it has been shown that the V protein is completely dispensable for viral replication in cell cultures but encodes a luxury function required for pathogenesis in vivo. The Sendai virus C proteins were also defined to be nonessential gene products which greatly contributed to replication both in vitro and in vivo. It is also now possible to design live vaccines by introducing predetermined or plausible attenuating mutations. In addition, the use of paramyxoviruses to express foreign genes has also become feasible. Paramyxovirus reverse genetics is thus renovating our understanding of viral replication and pathogenesis and will further mark an era in recombinant technology for disease prevention and gene therapy. PMID- 10386337 TI - Blood-borne virus infections in dialysis units--a review. AB - Hepatitis outbreaks in haemodialysis unit patients and staff were reported in the late 1960s. In 1972, the Rosenheim report in the UK established guidelines which included routine tests for hepatitis B surface antigen and isolation facilities for dialysing patients with hepatitis B virus which resulted in a dramatic fall in cases of hepatitis. However, since these guidelines were introduced, other blood-borne viruses, notably HCV and HIV have been discovered, and failures of infection control practices still lead to outbreaks of HBV in haemodialysis units. The prevalence of HCV in dialysis patients varies considerably throughout the world, with reported prevalence ranging from 3.9% to 71%. The number of blood transfusions and the length of time on dialysis have consistently been associated with HCV prevalence. Several reports provide evidence of patient-to-patient HCV transmission with environmental blood contamination the most significant factor in intra-unit transmission. There is no evidence that HCV has been transmitted by re-use of dialysis machines but being dialysed next to an HCV positive patient is associated with a significant risk of HCV acquisition. Several studies have shown that dialysing HCV positive patients in a separate unit or in a defined sector of a dialysis unit significantly reduces nosocomial HCV infection. HGV is prevalent in dialysis units where there is evidence of transmission to patients but no evidence of associated symptoms. HIV is infrequently transmitted in dialysis units and several units treating many HIV-positive patients have shown no evidence of transmission. Careful attention needs to be paid to infection control procedures and regular virological testing. PMID- 10386338 TI - The Nef protein of primate lentiviruses. AB - The Nef protein of primate lentiviruses acts as an important virulence factor in vivo both in monkeys and in humans. Among a human cohort of long-term non progressors, several Nef defective HIV1 viruses have been isolated, indicating that Nef may accelerate HIV progression and disease in humans. Additionally, a Nef-deleted SIV virus has low titres in rhesus monkeys and the animals develop AIDS at a much slower rate. In vitro, Nef can exert at least three kinds of effects: it downregulates CD4 and MHC class I, it stimulates virion infectivity and it alters signal transduction pathways. To accomplish this, Nef interacts with a series of cellular partners including CD4, components of the adaptor complexes AP-1 and AP-2, and several protein kinases, Nef often functioning as a connector between targets and effectors. The high degree of understanding of at least some aspects of Nef action, as well as the importance of this viral gene product for disease induction, identify Nef as a valuable target for the development of novel antiviral therapies. Moreover, the possibility of developing vaccines using attenuated viruses with deletions in nef and other crucial genes raises the possibility that the AIDS epidemic might one day be restrained. PMID- 10386339 TI - Immunisation strategies for viral diseases in developing countries. AB - In just under a quarter of a century, the Expanded Programme on Immunisation has been associated with an increase in infant immunisation coverage from around 5% to 80%, and the prevention of at least 3 million deaths annually, at very low cost. The global target of poliomyelitis eradication by the year 2000 appears feasible. Measles is the next likely target for eradication via immunisation, through 'catch-up', 'keep up' and 'follow-up' strategies which have proven highly effective in the Americas. Yet much needs to be done in order to extend readily achievable immunisation benefits equitably to all the world's people and to realise the potential of existing and soon to be available vaccines for disease control and eradication, as experience with yellow fever and hepatitis B vaccines demonstrates. Unsafe injection practices are widespread, have received inadequate attention, and cause a substantial global burden of blood-borne infections. The risk of increasing global inequity in immunisation highlights the centrality of resource allocation priorities in determining the extent to which the benefits of immunisation will be realised, particularly for new vaccines which are significantly more costly than established EPI vaccines. WHO/UNICEF strategies to target more effectively immunisation support to the neediest countries, to prioritise new vaccines, and to target carefully vaccine procurement and encourage sharply tiered vaccine pricing support both equity and sustainability. However, increasing the resources available to immunisation is vital and requires powerful advocacy on public health, moral, cost-effectiveness and legal grounds. More appropriate resource allocation priorities could readily provide the means necessary to address both technical and operational immunisation challenges. PMID- 10386340 TI - [Uniting basic and applied research]. PMID- 10386341 TI - Use of combined intraoperative radiotherapy and 125I brachytherapy in incompletely resected recurrent colorectal carcinoma. AB - This technique paper describes a new treatment strategy which involves the combination of 125I brachytherapy and intraoperative radiotherapy (IOERT) in the treatment of recurrent colorectal adenocarcinoma. IOERT is used to encompass the areas of presumed microscopic disease with the IOERT dose being kept below the threshold for severe neuropathy. Brachytherapy with 125I is used to boost areas of gross disease within the IOERT treated area that would otherwise require potentially neurotoxic IOERT doses to achieve local control. The outcome results of patients treated with this technique will be reported at a later date after further patient actual and longer follow-up. PMID- 10386342 TI - [Chronically administered midazolam does not have neurodegenerative effects]. AB - The effects of midazolam on the dorso- and ventromedial nuclei of the hypothalamus, administered during 120 days via gastric intubation, was studied in two groups of Wistar rats. The rats (50) of one of the groups were 2 months old, and those of the other (50) 24 months, 20 rats of both groups received 1 mg/kg of midazolam, and the other 20, 3 mg/kg As controls 20 rats of the same strain and age (10 for each group) received only saline. Neuronal count and karyometry did not revealed significant differences between controls and experimental rats. Only the group of old rats showed a slight increase in the number of dark neurons, with a decrease in the karyometric index. PMID- 10386343 TI - [New neuronal tracers and their combined use]. AB - Neuroanatomical tract-tracing methods are powerful tools for the study of brain circuits. The use of axonal tracers has become very popular during the past few years. Tract-tracing allows us to study the way in which two or more brain areas are connected and can be used to obtain detailed data on the processing of information within a particular area. The recent development of protocols combining several tracers has resulted in an important breakthrough. Although technically very demanding, these multitracer procedures have become state of the art protocols in several laboratories, rendering a broad range of possibilities for their application in Neurobiology. PMID- 10386345 TI - [Imaging a problem case]. PMID- 10386344 TI - [Regulation of cardiac output;an approximation at 3 levels: organic, cellular, and protein]. AB - The heart is the central point for adaptation of the organism to physical exercise because it is the center of the energy support system. Its activity is regulated at three levels; organ, cells and molecular and genetic components. During the development of the heart, the organ adapts in response to chronic and acute overloads by instantaneous functional and chronic changes, leading to a variable degree of cardiac growth. Physical exercise (acute and chronic) is the main example of physiologic overload. The acute response of the heart means a mechanical-hemodynamical and energetic modulation, driving to a final point where oxygen supply fits the increased need. Training, as response to chronic exercise, promotes an increase in energetic capacity (heart rate and stroke volume), structurally reflected in the physiological cardiac hypertrophy. Global functional and structural changes express what is happening at the cellular level. Different stimuli signal through specific receptors and second messengers to the nucleus, regulating gene expression and conditioning structural (size) and functional (contractile) changes. Changes in cellular size explain, by Starling mechanism, the increase in individual contractile strength and in reduction of the ventricular cavity in the systolic period. Other structural changes refer to the interstitium, myocardial vasculature and vascular reactivity. Changes in contractility affect the composition of the contractile elements (isoforms of heavy myosin, light myosin and/or modulatory proteins) and sarcoplasmic Ca2+ regulation, through the increase in Ca2+ flow. Many of the adaptations to chronic exercise studied in vivo in intact heart, isolated heart (Langendorf) or papillary muscle (multicellular preparation), are retained in the cardiomyocyte. Isolated cardiomyocytes can be precisely through the medium, temperature, ionic composition, active substances, etc. Shortening speed without load (Vmax), considered an inotropic index (Sonnenblick) can be measured independently of the initial length. Myocytes shorten against an internal load (restoration force) with viscous and elastic components, although they cannot be loaded externally (stretching is difficult). Cardiomyocyte isolation and maintenance requires strict and controlled conditions. This model offers many possibilities for studying dimensions, contraction-relaxation mechanics, Ca2+ and pH dynamics, beta adrenergic receptors, electrophysiology, pharmacology, genetics, etc. This kind of studies can deal with normal myocytes or myocytes from trained animals, cardiomyopathies, etc. PMID- 10386347 TI - [The doctor as seen by the literati]. PMID- 10386346 TI - [Trovafloxacin]. PMID- 10386348 TI - "The way we were". PMID- 10386349 TI - Update on permeation testing. PMID- 10386350 TI - Update on permeation testing. PMID- 10386351 TI - Investigation of bites and itching in a word processing department. AB - This study investigated arthropod and nonarthropod sources of reported bites and itching in a word processing division of a St. Louis, Mo., municipal department. Bird and rodent mites were suspected as causes of the bites because of the large population of pigeons around window ledges on some floors and the sighting of mice in the office. No mites or other arthropods were found to be responsible for the problem. Air samples were negative for fiber glass. Surface-vacuum samples collected around desks contained small quantities or traces of fiber glass or mineral wool. Humidity in the occupied space was considered low, about 35% relative humidity, with carbon dioxide measurements exceeding 1000 ppm. A single cause of the bites was not identified; however, a combination of surface-borne dust on desk tops and floors, the presence of minute quantities of mineral wool and fiber glass, relatively dry conditions, little or no outdoor air supplied to the work space, evidence of seasonally associated high work load, labor/management strife, and the presence of over 17 computers being used on a 24 hour basis (possibly leading to high levels of static electricity) were suspected as multiple causes of most of the "bites." After removal of loose mineral wool and dirt from an air handling unit and implementation of an aggressive cleaning routine, no more bites or itching were reported after a 6-month, 1- and 2-year follow-up period. Further research is needed to determine the relative importance of surface-borne dust and fibers, work stress, psychosocial support, and static electric fields, to produce bite-like sensations. PMID- 10386352 TI - Variabilities in aerosolizing activities and airborne fungal concentrations in a bakery. AB - Concentrations of airborne culturable fungi were measured in the kitchen of a bakery in Boston, Mass., to evaluate variabilities associated with common worker activities, outdoor aerosol distributions, and season. Activities were categorized as early morning preparation, cornmeal sifting and tossing, flour dumping and mixing, sweeping, and low activity. Sets of measurements were taken over 1 day in spring and 1 day in summer. Fungal concentrations were measured using a one-stage culture plate impactor, and bulk samples were taken from suspected fungal reservoirs within the bakery and subsequently cultured. Compared with the low activity category, elevated levels of total culturable fungi were found during all other activities, with the amount of increase closely related to individual worker activity as well as outdoor concentrations and initial bakery conditions. In the spring, Penicillium was the dominant genus showing activity related elevations in concentrations, while Cladosporium was the dominant genus during the summer. Clearly, due to variabilities in worker activities and ambient fungal concentrations, a standardized sampling protocol involving a large sample size over multiple days is needed to estimate accurately exposure to either total airborne fungi or specific fungal taxa. PMID- 10386353 TI - The use of composite dust wipe samples as a means of assessing lead exposure. AB - This study investigated two methods for analyzing composite dust wipes for lead. The term composite means two or more wipes collected from common components in a dwelling that are combined in the field and analyzed as a single sample. Two methods--a modified Environmental Protection Agency (EPA) Method 3050A and a Wisconsin Occupational Health Laboratory (WOHL) method--were selected based on their anticipated ability to handle the added mass of materials and dust expected in a composite. The study used off-the-shelf wipes to prepare single-, two-, and four-wipe samples. Wipes were spiked with a standard reference material at either a low dust loading level or a high level, and three laboratories analyzed the samples using both methods and both flame atomic absorption spectrometry and inductively coupled plasma-atomic emission spectrometry techniques (ICP). Good agreement with known spiked levels was possible using either method; the modified EPA 3050A showed particular promise. When up to four wipes were combined, all three laboratories found that modified EPA Method 3050A resulted in recoveries between 89 and 101% of the known standard. Although it was possible to achieve good agreement with spiked levels using the WOHL method, some difficulties were encountered, particularly when followed by ICP analysis and when using four wipes. The increased time required to digest the multiwipe composites was not proportional to the number of wipes in a composite: the two- and four-wipe composites did not take two to four times as long as a single-wipe sample. Laboratory analysis of a four-wipe sample cost an average of 65% less than analysis of four single-wipe samples for each method. PMID- 10386354 TI - Experimental deposition of environmental tobacco smoke submicrometer particulate matter in the human respiratory tract. AB - Measurements of 15 nonsmokers and 3 smokers breathing environmental tobacco smoke (ETS), were conducted to study particle deposition within the human respiratory tract. The subjects inhaled ETS of count median diameter (CMD) of about 0.2 micron and geometric standard deviation (GSD) of 1.7 The particle size distribution in the submicrometer range in the inhaled and exhaled air from the subjects was measured using a scanning mobility particle sizer (SMPS). A deposition of 56.0 +/- 15.9% was observed for nonsmokers while breathing ETS through the nose and 48.7 +/- 11.6% while breathing ETS through the mouth. One individual tested four times gave an average deposition of 57.4 +/- 11.5%, providing an indication of intraindividual variation. Such a variation is expected since the breathing rate was not controlled in order that an indication of the deposition experienced on a day-to-day basis could be obtained. For nonsmokers the deposition while breathing through the mouth was lower than through the nose and the variability within the measurements was also lower for mouth breathing. The latter could be due to the variation in individual size and shape of the nasal passage. Smokers had, on average, a higher rate of deposition but also a higher interindividual variability making it difficult to draw conclusions with respect to the affect of smoking on ETS particle deposition. The average deposition of the three smokers was 65.3 +/- 24.1% for nasal breathing and 66.1 +/- 17.6% for mouth breathing. PMID- 10386355 TI - Workplace use of an adjustable keyboard: adjustment preferences and effect on wrist posture. AB - This study presents an evaluation of an adjustable keyboard based on subjective preference and wrist joint motion during typing. Thirty-five computer users used the adjustable split design keyboard for 7-14 days during their usual work and were instructed to adjust the keyboard to the opening angle they preferred. At the end of this period, three-dimensional motion analysis was performed to compare the distribution of wrist joint angles while subjects typed on a conventional keyboard and the adjustable keyboard adjusted to the subject's preferred angle. The mean preferred opening angle was 14 degrees +/- 10. The mean ulnar deviation of the subjects who selected the opening angles between 21 and 28 degrees (n = 12) decreased from 18 degrees +/- 5 on the flat to 14 degrees +/- 5 on the adjustable (p < 0.05), while those who selected 0 to 10 degrees (n = 6) and 11 to 20 degrees (n = 17) split angles showed no significant differences in ulnar deviation. Mean wrist extension on the adjustable keyboard was 17 degrees +/- 5 and was significantly less than the 24 degrees +/- 5 observed on the conventional keyboard and most likely due to the presence of palm support. On average, subjects reported that the adjustable keyboard was more comfortable (0.5 +/- 0.5) (worse = -1, same = 0, better, = 1) in comparison with the conventional keyboard. PMID- 10386356 TI - Mineralogy of lung tissue in dental laboratory technicians' pneumoconiosis. AB - This article reports on a case of pneumoconiosis in a dental laboratory technician with a history of respiratory exposure to dental materials. Special attention is paid to the mineralogical analysis of the lung biopsy. The abundance of chromium, cobalt, and silica particles suggests that the dental technician's pneumoconiosis is the result of the combined effects of hard metal dusts and silica particles generated during finishing dental frameworks. Adequate technical protection such as a local ventilation system should be considered in dental laboratories to prevent respiratory exposure of dental technicians to airborne contaminants. PMID- 10386357 TI - Comparison of six respirator fit-test methods with an actual measurement of exposure in a simulated health care environment: Part III--Validation. AB - This article, the last in a series of three, describes the validation phase of a study conducted to test the correlation of respirator fit factors to the subject's actual exposure using biological sampling. The study consisted of three phases: protocol development, method comparison testing, and validation. Six quantitative fit-test methods were evaluated in the method comparison testing phase. The two fit methods with the highest correlation with the wearers' measured exposure were a corn oil method (R2 = 0.81) and an ambient aerosol method (R2 = 0.78). Because the ambient aerosol method is more commonly used in the workplace, it was selected for further analysis. In this validation phase, the fit factors measured during the ambient aerosol fit-test were used to calculate the exposures to Freon-113 by using the model determined in the method comparison testing phase of the study. The actual Freon-113 exposures were then measured and compared with the predicted exposures. The results verified that the ambient aerosol method fit factors are highly correlated to the total Freon-113 exposure dose and thus that the model had a predictive ability. PMID- 10386358 TI - Experimental investigation of power loss coefficients and static pressure ratios in an industrial exhaust ventilation system. AB - A study tested whether measures of equivalent resistance (X values) and ratios of static pressure (SPratio) for given ducts of contaminant control exhaust ventilation systems were independent of substantial changes to airflow level and to changes to resistance of other ducts within the same full-scale five-branch system. In a factorial study design, four airflow levels were achieved by changing fan rotation rate while resistances to flow for specific branch ducts were changed independently by adjusting slidegate dampers to various settings. For each damper insertion depth (including fully open), the results demonstrated substantial invariance for branch X values (few greater than 5%), SPratio (few greater than 3%), and fraction of airflow to each duct (few greater than 2%). X values for submains were much less stable, changing by 20% or more with changes to other parts of the system. For the same conditions, hood static pressures changed by as much as 96% (with standard deviation of 40%). The results suggest that before and after values of X and SPratios should be more reliable bases for indicating alterations than comparison of observed static pressures. The stability of airflow distributions with substantial changes in airflow suggests that one could adjust airflow distribution (e.g., with dampers) without considering whether the fan speed was set correctly, leaving fan adjustments for a final step. PMID- 10386359 TI - Exposures from thorium contained in thoriated tungsten welding electrodes. AB - Information provided in this article can be used for estimating the radiation dose associated with the use of thoriated tungsten electrodes in tungsten inert gas welding. Area and breathing zone concentrations of 232Th generated by welding and electrode sharpening along with particle size information; isotopic composition of electrodes from two domestic manufacturers and one European manufacturer; and process variables and estimates on the number of thoriated tungsten electrodes manufactured are presented. Past literature is reviewed and compared with the results of this study. Isotopic analysis of a nominal 2% thoriated electrode found 0.6 ppm +/- 0.4 ppm 230Th and less than 0.1 ppm 228Th. Analysis of a ceriated tungsten electrode and a lanthanated tungsten electrode for 232Th found 124 ppm and 177 ppm, respectively. Electrode consumption during welding was primarily the result of tip sharpening. Less than 3% of the weight loss was attributable to the welding process. The in-mask concentration of respirable thorium particulate in the welder's breathing zone was 0.002 x 10(-12) microCi 232Th/mL. The concentration of respirable thorium particulate from electrode sharpening was 1.3 x 10(-12) microCi 232Th/mL. The measured sharpening time was 20 sec per electrode. Estimates of the activity median aerodynamic diameters for the respirable fraction of the welding and electrode sharpening aerosols were 3.5 and 5 microns, respectively, when measured in the breathing zone at 0.3 m (12 inches) from the point of operation. The respirable fraction of the total welding and sharpening aerosols was 45 and 60%. PMID- 10386360 TI - An ergonomic analysis of premixing and compounding processes in an animal health plant. AB - This study is based on an ergonomic job analysis designed to develop a hazard prevention program for the premixing and compounding processes in animal health products plants. Animal health products plants are "pharmaceutical facilities" for livestock or domestic animals. A hazardous aerosol (highly concentrated antibiotics, anthelmintics, mineral oil, and animal hormones) is generated in the premixing or compounding processes. The animal health premixing jobs are heavy duty jobs and have high potential for chemical exposure, heat stress, and ergonomic hazards. Ergonomic job analysis was used to recognize, identify, and evaluate actuarial and potential risks of injures or irritations. Chemical hazards and eight ergonomic factors were discussed: (1) forceful exertions, (2) awkward postures, (3) localized contract stresses, (4) vibration, (5) noise, (6) temperature extremes, (7) repetitive activities, and (8) prolonged activities. The results show that (1) current practices do not violate occupational safety and health regulations or recommended guidelines, but that hazards should be identified to protect worker health and safety; (2) for chemical hazards prevention, operators wear whole-body protection equipment, which also causes heat stress and increases the noise level in the work zone; and (3) the loading weight of the bags used needs to be reduced. PMID- 10386361 TI - Issues and controversy: the measurement of crystalline silica; review papers on analytical methods. AB - This article reviews the analytical methods for crystalline silica polymorphs and summarizes promising techniques for compliance with health-related regulations. X ray diffraction analysis appears to be the most promising method of determining quartz and cristobalite content at this threshold in many bulk mineral and chemical systems. Other analytical techniques can be used in some mineral and chemical assemblages, but usually lack polymorph specificity or sensitivity. All analytical methods benefit from concentration techniques that do not alter crystalline properties of silicas. National Institute of Occupational Safety and Health (NIOSH) Analytical Method 7601 as written suffers from destruction of the mineral residue containing crystalline silica polymorphs after dissolution of many silicate minerals in phosphoric acid and may also include digestion resistant silicate minerals, but it is a promising concentration method for other analytical methods such as NIOSH Analytical Method 7500 (X-ray diffraction). PMID- 10386362 TI - Simulated blood levels of CF3I in personnel exposed during its release from an F 15 jet engine nacelle and during intentional inhalation. AB - Of the agents under consideration for protecting unoccupied areas from fire, CF3I (trifluoroiodomethane) has physicochemical properties that give it potential as a "drop-in" replacement for halon 1301. One of the issues concerning the use of CF3I is the potential hazard to ground crews should an inadvertent discharge occur while workers are in or near an engine nacelle. A discharge test of CF3I was conducted on an F-15A jet to record CF3I concentration time histories at locations near the aircraft. The conditions of the discharges simulated an inadvertent ground discharge with the engine nacelle doors open and also with the doors closed. The use of three types of gas analysis instrumentation allowed gas sampling from several locations during the discharge tests. Concentrations measured at selected sensor locations were used as the input to a physiologically based pharmacokinetic model to simulate blood levels that would be attained by individuals inhaling CF3I at sensor locations. Blood levels reached during these exposures were compared with the blood level associated with the lowest observable adverse effect level (LOAEL) for cardiac sensitization to evaluate the possibility of safe egress. The highest blood concentrations simulated were twice the target blood concentration associated with cardiac sensitization. However, simulated blood concentrations of subjects who actually inhaled CF3I reached levels that were 100 times the target level without reported adverse effect. Thus, actual human data may supersede the use of the cardiac sensitization LOAEL obtained from animal studies. PMID- 10386363 TI - Whole-body vibration of locomotive engineers. AB - Vibration of the seat and the body of a diesel locomotive and an electric locomotive were measured while driving on the railways of Eastern Finland. At the speed of 120 km/h for the diesel locomotive and 140 km/h for the electric locomotive (the greatest permissible speeds) the vibration of the seat was tangent to the "fatigue-decreased proficiency boundary" of the international standard ISO 2631/1(1) in the side-to-side direction in one-third octave bands of 1-1.6 Hz. The frequency response measurements between the body of the locomotive and the seat indicated that the seat did not reduce side-to-side vibration of the body at low frequencies where the vibration was most harmful. PMID- 10386364 TI - Molecular biology of mycotoxin biosynthesis. AB - Mycotoxins are secondary metabolites produced by many important phytopathogenic and food spoilage fungi including Aspergillus, Fusarium and Penicillium species. The toxicity of four of the most agriculturally important mycotoxins (the trichothecenes, and the polyketide-derived mycotoxins; aflatoxins, fumonisins and sterigmatocystin) are discussed and their chemical structure described. The steps involved in the biosynthesis of aflatoxin and sterigmatocystin and the experimental techniques used in the cloning and molecular characterisation of the genes involved in the pathway are described in detail. The biosynthetic genes involved in the fumonisin and trichothecene biosynthetic pathways are also outlined. The potential benefits gained from an increased knowledge of the molecular organisation of these pathways together with the mechanisms involved in their regulation are also discussed. PMID- 10386365 TI - Regulation of the transcription of genes encoding different virulence factors in Helicobacter pylori by free iron. AB - Since free iron possesses a poor solubility under physiologic conditions and thus becomes a limiting nutrient for growth, a shift from high- to low-iron environmental conditions is an important signal for bacteria to coordinate the regulation of gene expression. Here, we studied and compared the level of transcripts corresponding to the vacA (cytotoxin), ureA (urease), cagA (cytotoxin associated antigen) and fur (ferric uptake regulator) genes of Helicobacter pylori, grown under iron-sufficient and iron-restricted conditions. A significant increase in the accumulation of vacA and fur transcripts was observed under iron restricted conditions. This up-regulation by low levels of iron seems to be not directly regulated by Fur, and certainly requires other regulatory factors. No statistical difference was defined in the accumulation of cagA and ureA. PMID- 10386366 TI - Nucleotide sequence of the streptococcin A-FF22 lantibiotic regulon: model for production of the lantibiotic SA-FF22 by strains of Streptococcus pyogenes. AB - Streptococcin A-FF22 (SA-FF22) is a type AII linear lantibiotic produced by Streptococcus pyogenes strain FF22. Sequence analysis of an approximate 10 kb region of DNA showed it to contain nine open reading frames arranged in three operons responsible for regulation, biosynthesis and immunity of SA-FF22. This region is organized similarly to the Lactococcus lactis lacticin 481 region, however, unlike lacticin 481, a two-component regulatory system is essential for SA-FF22 production. Located immediately downstream of the scn region is a putative transposase gene, the presence of which supports earlier data that indicated a mobile nature to this region. PMID- 10386367 TI - Isolation of the murI gene from Brevibacterium lactofermentum ATCC 13869 encoding D-glutamate racemase. AB - The murI gene encoding D-glutamate racemase plays an important role in the biosynthesis of D-glutamic acid, an essential component of cell wall peptidoglycan of almost all eubacteria. A DNA fragment that could rescue the auxotrophy of D-glutamic acid in the Escherichia coli murI mutant strain WM335 was isolated from Brevibacterium lactofermentum ATCC 13869 belonging to the coryneform bacteria. DNA sequencing reveals that it encodes a protein of 284 amino acid residues, which shows a high level of homology with D-glutamate racemases from several other bacteria. PMID- 10386368 TI - Deletions in the repeating sequences of the toxin A gene of toxin A-negative, toxin B-positive Clostridium difficile strains. AB - The repeating sequences of the toxin A gene from toxin A-negative, toxin B positive (toxin A-, toxin B+) strains of Clostridium difficile which were isolated in geographically separated facilities in Japan and Indonesia were determined. All six strains tested had identical repeating sequences with two deletions (1548 and 273 nucleotides in size) in the toxin A gene. A PCR method was designed to detect the deletions and the deletions were confirmed in all 50 toxin A-, toxin B+ strains examined by this method. Western immunoblot analysis revealed that polyclonal antiserum against native toxin A did not react with the concentrated culture filtrates of the toxin A-, toxin B+ strains. These results may suggest that toxin A-, toxin B+ strains have deletions of the two thirds of the repeating regions of the toxin A gene, which encodes the epitopes fully responsible for the reaction with the polyclonal antiserum. PMID- 10386369 TI - Purification and characterization of triheme cytochrome c7 from the metal reducing bacterium, Geobacter metallireducens. AB - A soluble c-type cytochrome was first purified from Geobacter metallireducens to an electrophoretically homogeneous state. The purified cytochrome c showed absorption peaks at 530 and 409 nm in the oxidized form and 552, 522, and 418 nm in the reduced form. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate allowed us to calculate the molecular mass at 9.5 kDa. It contained 3 mol of heme c per molecule of the protein on the basis of heme c and protein concentration. The mid-point redox potential at pH 7.0 was determined to be -190 mV. Although the N-terminal amino acid sequence of the first 17 residues was similar to that of Desulfuromonas acetoxidans cytochrome c7, G. metallireducens cytochrome c did not show Fe(III)-reducing activity. PMID- 10386370 TI - Inhibitory effect of lipopolysaccharide on apoptotic cell death in macrophages infected with Actinobacillus actinomycetemcomitans. AB - We have reported that the macrophage-like cell line J774.1, when infected with the periodontopathic bacterium Actinobacillus actinomycetemcomitans, undergoes apoptosis. In this study, we examined whether stimulation of J774.1 cells with lipopolysaccharide (LPS) before the infection affects the subsequent apoptosis. Cytotoxicity on the LPS-stimulated cells was about half of the unstimulated control cells. DNA fragmentation in the LPS-stimulated cells was also significantly lower than in the control cells, whereas it was increased to a level similar to that of the control cells by addition of a nitric oxide (NO) inhibitor. In addition, significantly smaller numbers of live A. actinomycetemcomitans were recovered from the LPS-stimulated macrophages at 8 h after the infection as compared with the control cells. These findings suggest that the inhibitory effect of LPS on apoptosis results from an enhanced NO mediated bactericidal activity. PMID- 10386371 TI - Mercaptopyridine-N-oxide, an NADH-fumarate reductase inhibitor, blocks Trypanosoma cruzi growth in culture and in infected myoblasts. AB - The enzyme NADH-fumarate reductase is not found in mammalian cells but it is present in several parasitic protozoa including Trypanosoma cruzi, the parasite that causes Chagas' disease. This study shows that the drug 2-mercaptopyridine-N oxide (MPNO) inhibits NADH-fumarate reductase purified from T. cruzi (ID50 = 35 microM). When added to intact cells, MPNO inhibited the growth of T. cruzi epimastigotes in culture (ID50 = 0.08 microM) as well as the infection of mammalian myoblasts by T. cruzi trypomastigotes (ID50 = 20 microM). At a concentration of 2.4 microM, MPNO also inhibited the growth of amastigotes (intracellular dividing forms) in cultured mammalian myoblasts. Supplementation of culture media with 5 mM succinate, the product of fumarate reductase, partially protected against the inhibition of the growth of epimastigotes by MPNO. Moreover, MPNO inhibited the accumulation of succinate in cultures of epimastigotes, as measured by high performance liquid chromatography. Although MPNO may have other intracellular targets in addition to fumarate reductase, these results support the hypothesis that compounds which inhibit the enzyme fumarate reductase may be potential chemotherapeutic agents against Chagas' disease. PMID- 10386373 TI - Chromosome mapping in Cryptosporidium parvum and establishment of a long-range restriction map for chromosome VI. AB - We used contour-clamped homogeneous electric field (CHEF) gel electrophoresis and Southern blot hybridization to analyze the molecular karyotype of Cryptosporidium parvum and establish the chromosomal location of 12 single copy genes. In agreement with previous studies, the molecular karyotype of C. parvum was found to consist of partially co-migrating chromosomes ranging in size from 0.97 to 1.55 Mb and segregating into five distinct electrophoretic bands. Hybridization results allowed the definition of a linkage group comprised of five distinct loci located on chromosome VI. Southern hybridization and restriction analysis of total C. parvum chromosomes or isolated chromosome VI using gene-specific probes and an oligonucleotide specific for C. parvum telomeres allowed the development of a long-range restriction map of chromosome VI. PMID- 10386372 TI - IST1 insertional inactivation of the resB gene: implications for phenotypic switching in Thiobacillus ferrooxidans. AB - Thiobacillus ferroxidans ATCC 19859 undergoes rapid phenotypic switching between a wild-type state characterized by the ability to oxidize ferrous iron (FeII) and reduced sulfur compounds and a mutant state where it has lost the capacity to oxidize FeII but retains the ability to oxidize sulfur. The mutant has also gained the capacity to swarm. It is proposed that loss of FeII oxidation is due to the reversible transposition of the insertion sequence IST1 into resB encoding a putative cytochrome c-type biogenesis protein. Downstream from resB and co transcribed with it is resC, encoding another putative cytochrome biogenesis protein. IST1 insertional inactivation of resB could result in the loss of activity of its target c-type cytochrome(s). This putative target cytochrome(s) is proposed to be essential for FeII oxidation but not for sulfur oxidation. Curiously, resB and resC pertain to the proposed system II cytochrome biogenesis pathway whereas gamma Proteobacteria, of which T. ferrooxidans is a member, normally use system I. This could represent an example of lateral gene transfer. PMID- 10386374 TI - Structure and expression properties of the endo-beta-1,4-glucanase A gene from the filamentous fungus Aspergillus nidulans. AB - Endo-beta-1,4-glucanase A (EG A) of Aspergillus nidulans was purified to homogeneity, and its genomic gene (eglA) was cloned based on partial amino acid sequences of the purified enzyme and sequenced. The eglA gene comprised 1228 bp with four putative introns and encoded a polypeptide of 326 amino acids bearing high homology to the family A cellulases. The eglA promoter activity in A. nidulans was examined using the A. oryzae Taka-amylase A gene as a reporter. Expression of the reporter gene was induced by carboxymethylcellulose and cellobiose, and repressed by glucose, galactose, mannose, xylose, sorbitol, glycerol and succinate. Lactose neither induced nor repressed the expression. PMID- 10386375 TI - Non-curliation of Escherichia coli O78:K80 isolates associated with IS1 insertion in csgB and reduced persistence in poultry infection. AB - The elaboration of curli fimbriae by Escherichia coli is associated with the development of a lacy colony morphology when grown on colonisation factor antigen agar at 25 degrees C. Avian colisepticaemia E. coli isolates screened for curliation by this culture technique showed lacy and smooth colonial morphologies and the genetic basis of the non-curliated smooth colonial phenotype was analysed. Two smooth E. coli O78:K80 isolates possessed about 40 copies of the IS1 element within their respective genomes of which one copy insertionally inactivated the csgB gene, the nucleator gene for curli fibril formation. One of these two isolates also possessed a defective rpoS gene which is a known regulator of curli expression. In the day-old chick model, both smooth isolates were as invasive as a known virulent O78:K80 isolate as determined by extent of liver and spleen colonisation post oral inoculation but were less persistent in terms of caecal colonisation. PMID- 10386376 TI - Green fluorescent protein as a detection marker for Coxiella burnetii transformation. AB - The molecular biological study of the obligate intracellular bacterium Coxiella burnetii is hampered because of the lack of an efficient DNA transformation system. We used expression of the green fluorescent protein (GFP) in addition to ampicillin resistance as a selection marker for detection of transformed C. burnetii cells. Fluorescent microscopy studies revealed that transformed C. burnetii cells can be detected easily inside the host cell line. A high level of GFP expression was reached with the strong Escherichia coli trc (trp/lac) promoter. The use of GFP not only provides a convenient marker for transformation of C. burnetii, but also allows detection of this obligate intracellular pathogen inside host eukaryotic cells. Possible applications for GFP in the study of host pathogen interactions are discussed. PMID- 10386377 TI - Development of a new PCR-ribotyping method for Clostridium difficile based on ribosomal RNA gene sequencing. AB - PCR-ribotying, a typing method based on polymorphism in the 16S-23S intergenic spacer region, has been recently used to investigate outbreaks due to Clostridium difficile. However, this method generates bands of high and close molecular masses which are difficult to separate on agarose gel electrophoresis. To improve reading of banding patterns of PCR-ribotyping applied to C. difficile, a partial sequencing of the rRNA genes (16S and 23S) and intergenic spacer region has been performed, then a new set of primers located closer to the intergenic spacer region has been defined. The new PCR gave reproducible patterns of bands easy to separate on agarose gel electrophoresis. Each of the 10 serogroups and 11 subgroups of serogroup A produced a different pattern. This typing method has evidenced major qualities such as easiness, rapidity and reproducibility. However, its discriminatory power has to be evaluated to validate its importance as a typing tool for C. difficile. PMID- 10386378 TI - Specific interactions between Porphyromonas gingivalis fimbriae and human extracellular matrix proteins. AB - The interactions of the extracellular matrix (ECM) proteins (laminin, elastin, fibronectin, type I collagen, thrombospondin and vitronectin) with the fimbriae of Porphyromonas gingivalis were analyzed based on surface plasmon resonance (SPR) spectroscopy using a biomolecular interaction analyzing system (BIAcore). The BIAcore profiles demonstrated that fimbriae specifically bound to all of the ECM proteins with significant association constants (Ka). Vitronectin showed the highest affinity to fimbriae (Ka = 3.79 x 10(6) M-1), while the affinity of laminin was lowest (Ka = 2.15 x 10(6) M-1). A synthetic peptide which is a potent inhibitor of fimbrial binding to salivary proteins was not significantly effective on the fimbrial interactions with the ECM proteins. Using polystyrene microtiter plates revealed that P. gingivalis fimbriae bound markedly to immobilized fibronectin and type I collagen, while the interaction of fimbriae with the other ECM proteins was not clearly demonstrated. These results suggest that interactions between fimbriae and the ECM proteins occur with specific affinities which are not mediated by mechanisms identical to those of salivary proteins. It was also shown that SPR spectroscopy is a useful method to analyze these specific interactions. PMID- 10386379 TI - Partial nucleotide sequencing of the NS3/helicase region of hepatitis G virus to prove vertical transmission. AB - To study non-parental transmission of hepatitis G virus and/or GB virus C (HGV/GBV-C), we sequenced and compared the NS3/helicase region of the virus for five HGV/GBV-C RNA-positive mothers and their 11 children who had experienced neither blood transfusion nor overt hepatitis and were negative for HBV, HCV and HIV, except in one mother coinfected with HCV. The nucleotide sequences of the familial HGV/GBV-C isolates showed high similarity of 99-100% (mean 99.8%, 100% at the deduced amino acid level) between mother and her child(ren) in each family. These findings strongly suggest the spontaneous occurrence of mother-to child transmission of HGV/GBV-C as reported previously. They also suggest that nucleotide sequence analysis on the NS3/helicase region of HGV/GBV-C may be a useful tool to study HGV/GBV-C transmission. PMID- 10386380 TI - Physical and genetic map of the Listeria monocytogenes EGD serotype 1/2a chromosome. AB - Listeria monocytogenes is a facultative intracellular pathogen responsible for both invasive and non-invasive food-borne illness in animals and humans. In this study, macrorestriction analysis following pulsed-field gel electrophoresis was used to show that Listeria monocytogenes serovar 1/2a strain EGD has a single chromosome containing eight NotI fragments of 1100, 850, 365, 320, 275, 40, 30 and 20 kb in size and 11 AscI fragments of 860, 470, 410, 360, 320, 250, 110, 80, 50, 30 and 20 kb. The total genome therefore comprises 3000 +/- 50 kb. The creation of a physical and genetic map of the Listeria genome was achieved by generating NotI linking clones and their use in subsequent hybridisation analysis. Using isogenic mutants harbouring additional artificial NotI restriction sites, we were able to precisely map the positions of all currently known virulence genes on the chromosome. PMID- 10386381 TI - Isolation and expression of a nitrogen regulatory gene, nmc, of Penicillium roqueforti. AB - The nmc gene, encoding a global nitrogen regulator, has been cloned and characterized from Penicillium roqueforti, a fungus used in the dairy industry. The deduced amino acid sequence predicts a protein of 860 amino acids in length whose zinc finger DNA binding domain is at least 94% identical to those of the homologous fungal proteins. Northern blot analysis showed that nmc expression is induced by nitrogen starvation and not repressed by variation of the external pH. PMID- 10386382 TI - Olfactory cues associated with the major histocompatibility complex. AB - Besides its immunological function of self/non-self discrimination the major histocompatibility complex (MHC) has been recognized as a possible source of individual specific body odors. Dating back to speculations on the role of the extraordinary polymorphism of the MHC as background of an individual chemosensory identity and to early observations of MHC-dependent mate choice in inbred strains of mice, systematic experimental studies revealed a first evidence for H-2 related body odors in this species. Meanwhile a large number of animal studies with rodents and a series of field studies and experiments with humans have extended our knowledge of MHC-related odor signals and substantiated the hypothesis of immunogenetic associated odor types. These results suggest that the most prominent feature of the MHC, its extraordinary genetic diversity, seems in part to be selectively maintained by behavioral mechanisms which operate in contemporary natural populations. The high degree of heterozygosity found in natural populations of most species seems to be promoted by non-disease-based selection such as mating preferences and selective block of pregnancy. PMID- 10386383 TI - Developmental aspects of kin recognition. AB - The ability to recognise kin requires the individual to possess a variety of abilities. Individuals must produce a cue which indicates relatedness, they must process this cue to determine relatedness and then must act on this cue. Research has concentrated on the first and final stage of this process, i.e., the cues of kinship and kin correlated behaviour. Little attention has been paid to how individuals process cues to determine relatedness. This paper discusses how individuals 'recognise' kin, the exhibition of kin correlated behaviour and considers the role of the MHC in these processes. Two broad theories have emerged to explain how individuals recognise their kin: either a recognition gene(s) or some experiential mechanism. In mammals there is no evidence to suggest that recognition (the processing of the cue) is under genetic control but rather is the result of experience, learning about related individuals during development. Moreover studies on kin recognition in the domestic dog suggest that all kin are not recognised by the same process but different classes of kin, parents, siblings may well be recognised using different means. Studies of kin correlated behaviour suggest that the behaviour exhibited towards kin by Mongolian gerbils is mediated by the environment. Factors of environmental familiarity, sex and developmental age all affect the response of individuals to kin and non-kin. In these situations the ability to recognise kin does not change but the exhibition of kin correlated behaviour changes according to environmental conditions. These studies indicate that kin recognition may not be the 'unified' process previously thought and thus any explanations of the proximate and ultimate causation of kin recognition need to encompass this complexity. The question remains of whether the MHC is complex enough to do so. PMID- 10386384 TI - Balancing selection and MHC. AB - The MHC is highly polymorphic in most vertebrates and the suggested selective mechanisms responsible for the maintenance of this variation are several, including maternal-fetal interaction, parasite resistance, and negative assortative mating. Evidence for these mechanisms is reviewed and estimates of the amount of selection in a number of studies are given. Although there is much yet to be understood about the mechanism and extent of balancing selection at MHC, new advances in molecular genetic technology and increasing interest in MHC from many types of biologists promise answers in the near future. PMID- 10386385 TI - The new heterozygosity theory of mate choice and the MHC. PMID- 10386386 TI - Soluble MHC antigens and olfactory recognition of genetic individuality: the mechanism. PMID- 10386387 TI - The present status of the 'carrier hypothesis' for chemosensory recognition of genetic individuality. AB - The hypothesis that soluble MHC Class I molecules could act as the vector which transports MHC-specific odours from the blood into the urine was put forward some ten years ago (Singh, Brown & Roser, 1987). Here, I summarise new evidence in favour of the hypothesis. 'We propose that the ability of MHC Class I molecules to associate in a selective way with other small molecules could also be the mechanism by which a unique mixture of volatile, endogenous metabolites is transported by [soluble] class I MHC glycoproteins from the blood into the urine' (Singh, Brown & Roser, 1987). PMID- 10386388 TI - Origin, functions and chemistry of H-2 regulated odorants. AB - Genes located within the major histocompatibility complex (MHC) of mice are responsible for individual differences in body odor (odor types). In this review we suggest that the MHC genes themselves are responsible for odor differences among MHC-congenic mice. Recent studies indicating that volatile carboxylic acids are at least in part responsible for the individual odors and what this finding implies about the pathway from gene to odorant are also reviewed. We suggest that odorants or their precursors are bound directly by MHC products and are released into serum and concentrated in urine. Finally, possible functions of MHC odor types in mice are enumerated and important future research questions are raised. PMID- 10386389 TI - A review of MHC-based mate preferences and fostering experiments in two congenic strains of mice. AB - We review our studies of mate choice with two MHC-congenic strains of mice. This work was stimulated by findings from Yamazaki and colleagues showing that male mice exhibited mate preferences for females whose MHC-haplotype was different from their own, while female mice exhibited either no preference or a weak preference for males of a particular MHC-haplotype (see Beauchamp et al., 1988). Since these findings were unexpected (mate choice theory predicts that females will be more selective than males), we studied the preferences of mice from two additional MHC-congenic strains to assess the generality of the previous findings. Specifically, the goals of our research were: (1) to determine the mate preferences of congenic mice with MHC-haplotypes derived from wild populations, (2) to compare the mate preferences of male and female mice in a test situation where each sex has a clear opportunity to make a choice, and (3) to estimate effects of cross-fostering on each sex. PMID- 10386390 TI - Use of the MHC for mate choice in wild house mice (Mus domesticus). AB - The mechanisms maintaining natural diversity at the major histocompatibility complex (MHC) are not well understood. To increase knowledge of one potential mechanism, I examined the use of MHC genes for mate choice by wild house mice in a controlled laboratory setting. Three rearing groups of wild test mice were produced: non-fostered control mice, mice fostered into families of an inbred laboratory mouse strain, and mice fostered into families of a second, MHC congenic mouse strain. Mature test mice were given a choice of two opposite-sex stimulus mice from the two MHC-congenic strains used for fostering, and were scored for several measures of preference. The results were non-significant in general, but females of two rearing groups spent significantly more time with mice of one MHC-type, and in most rearing groups, mice tended to spend more time with this same MHC-type. Other results showed that male test mice ejaculated indiscriminantly and that female wild mice mated to ejaculation more often in longer length trials, but showed no significant preferences. In this study, fostering seemed to have little or no effect on MHC-based mate preferences of wild house mice, and wild mice did not appear to be using the MHC to avoid inbreeding. However, some wild female mice used the MHC to choose potential mates. PMID- 10386391 TI - Behavioural studies of MHC-congenic mice. AB - Behavioural studies of MHC-congenic mice and rats have focused primarily on mate choice and the ability to discriminate between strains by their urine odours, but these strains may differ in other behaviours, such as activity and ultrasonic vocalizations. Ivanyi (1978, Proc. Roy. Soc. Lord. 202, 117-158) has reviewed the physiological differences associated with the MHC, many of which could influence behaviour. We have started a systematic study of behavioural development and adult behaviour in MHC-congenic mice. A developmental test battery (growth, rate, locomotion, grooming, eye opening, ultrasonic vocalizations, etc.) was used to examine differences between C57BL/6J vs. B6-H-2bml and C57BL/10SnJ vs. B10.BR/sgSnJ mice. A test battery of spontaneous behaviours (activity, exploration, ultrasonic vocalizations, etc.) was used to examine behavioural differences between adult C57BL/6J vs. B6-H-2bml; and C57BL/10SnJ vs. B10.BR/sgSnJ mice. Differences in development and in adult behaviours between these MHC-congenic strains is discussed in relation to possible neural, endocrine and immune system differences. Future studies will compare MHC-congenic mice on levels of anxiety, sociosexual behaviour and on learning paradigms. PMID- 10386392 TI - Olfaction and human kin recognition. AB - Humans, like other mammals, are capable of discriminating between kin and non-kin by olfactory cues alone. Shortly after birth, breastfed infants become familiar with, and respond preferentially to, their mother's unique odor signature. Mothers likewise recognize the characteristic scent of their newborn infant. Close biological relatives share somewhat similar odor signatures (presumably resulting from genetically mediated similarities in bodily biochemistry and metabolism) that could facilitate kin recognition. PMID- 10386393 TI - The major histocompatibility complex and the chemosensory signalling of individuality in humans. AB - The chemosensory identity of mice and rats is determined partly by polymorphic genes of the major histocompatibility complex (MHC). In inbred strains of mice, as well as in seminatural populations, MHC-associated mating preferences selectively influence reproductive success, thus serving to promote heterozygocity in the MHC. In order to determine whether MHC-associated chemosignals are present in humans, two studies were conducted. In a first study, olfactory identification of MHC-associated chemosignals was conducted on 12 trained rats' responses to the urine odors of humans. In a second study, MHC associated olfactory cues in humans were analyzed by means of gas chromatography. The results indicate that the urine odors of humans are associated with the MHC and demonstrate that the profile of volatile components in the urine odors shows some association with the MHC. Furthermore, results show that a profile of some specific components, as well as a few ubiquitous volatiles, constitutes MHC associated odor signals in humans. PMID- 10386394 TI - Molecular forms of soluble HLA in body fluids: potential determinants of body odor cues. AB - The major histocompatibility complex (MHC) has been linked to encoding for individual olfactory identity. Experiments in mice and rats proved that behavior and mating were, at least in part, determined by genes within the MHC. This study was aimed at investigating whether sHLA are excreted in human urine, saliva and sweat. In particular examination of the molecular forms in these fluids would give clues to whether break down forms of soluble MHC molecules might participate in shaping behavior. Major bands of 45, 40, and 23 kD were detectable. Increased levels of sHLA were measured using a quantitative ELISA in urine shortly before ovulation decreasing to normal levels thereafter. In animal models strain specific MHC-linked odor cues have been detected in urine. Thus, excretion of sHLA in urine might indicate a similar role for these molecules in humans. PMID- 10386395 TI - Body odor evoked potentials: a new method to study the chemosensory perception of self and non-self in humans. AB - A new method will be presented which allows the perception of body odors in humans to be studied objectively. The analysis of body odor-evoked potentials was used to investigate if and how the human brain is able to differentiate self from non-self body odor for the first time. Six subjects (three females) participated in two experimental sessions. In each session, two body odors (axillary hair) were presented within an olfactory oddball paradigm. One of the odors was collected from the subject and the other from an odor donor of the same sex. In the first session the subjects' attention was distracted to a secondary task (passive paradigm), in the second session the subjects were asked to actively differentiate the odors (active paradigm). For the EEG recordings the odors were presented within a constantly flowing airstream. The results show that the subjects could hardly differentiate the body odors subjectively. However, it could be demonstrated that the central nervous processing of one's own odor was faster than the processing of the chemosensory non-self signal. Moreover, in the active paradigm, the potentials appeared to be larger when the subjects perceived their own body odor. The conclusion is reached that the measurement of chemosensory event-related potentials (CSERP) is the method of choice for the investigation of HLA-associated body odors. PMID- 10386396 TI - MHC variation in birds and reptiles. AB - The major histocompatibility complex (MHC) has been studied in a multitude of mammals by now, but much less is known about its organisation and variation in other vertebrate species. The mammalian MHC is organised as a single gene cluster, but recent studies on birds suggest that this paradigm of MHC organisation has to be supplemented. The domestic chicken thus possesses two separate gene clusters which both contain MHC class I and class II B genes, and we have shown that the ring-necked pheasant Phasianus colchicus also has two unlinked clusters of class II B genes. We are studying the effect of the MHC on mate choice, survival and reproductive success in natural populations of birds and reptiles. For this reason, we are developing DNA techniques to determine the animals' MHC genotype. The amplification of the hypervariable exon 3 of the class I gene from songbirds and reptiles has provided us with species specific probes that can be used in Southern blot analysis. The first results indicate very extensive variation in all studied species, that is starlings Sturnus vulgaris, great reed warblers Acrocephalus arundinaceus and water pythons Liasis fuscus. The restriction fragment length polymorphism (RFLP) analysis also suggests that the number of MHC genes is significantly larger in these species than in pheasants and domestic chickens. PMID- 10386397 TI - Conservative treatment of the aortic root in acute type A dissection. AB - OBJECTIVE: In acute type A dissection long-term results of conservative aortic root surgery were compared with the outcome of primary valve and/or root replacement. METHODS: Between 1985 and 1995, 199 patients (mean age 59 years, 154 men) were operated on. The aortic root was involved in the dissection process and valve incompetence of varying degree was present without exception. Replacement of a proximal aortic segment was standard procedure in all patients. The aortic valve was preserved in 126 patients: commissural suture resuspension (12 patients), root reconstruction with GRF-glue (gelatine-resorcin formaldehyde/glutaraldehyde-glue) (114 patients). Valve replacement was performed in 73 patients (50 composite grafts, 23 valve prostheses with separate supracoronary grafts). Preoperative risk factors (valve replacement vs. preservation): coronary artery disease (11 vs. 8%, NS), tamponade (18 vs. 17%, NS), unstable hemodynamics (22 vs. 15%, NS), renal failure (4 vs. 6%, NS), neurologic disorder (19 vs. 32%, NS). RESULTS: The overall early mortality was 23.6% (47/199 patients) and increased after commissural suture resuspension compared with GRF-glue reconstruction (P = NS). Parameters of the early postoperative period did not differ between conservative treatment and root/valve replacement: low cardiac output, 34 versus 38% (P = NS); myocardial infarction, 10 versus 11% (P = NS); hemorrhage, 25 versus 23% (P = NS); duration of intensive care (P = NS). Survival was 61% after 8 years without difference between the two principal treatment groups (P = NS) and between the two conservative subgroups (P = NS). At 2 years, GRF-glue reconstruction had an increased freedom from reoperation on the aortic root (92 vs. 70%, P = 0.0253) and event free survival (77 vs. 41%, P = 0.0224) compared with suture resuspension. Commissural suture resuspension was an independent, significant predictor for reoperation (P = 0.0221, relative risk = 4.7130). CONCLUSION: Surgery for acute type A dissection still carries a considerable early risk. Preservation of the aortic root is safe in the absence of Marfan or annuloaortic ectasia, but a certain incidence of reoperations on the aortic valve and the aortic root has to be accepted. Root reconstruction using GRF-glue is the method of choice and is superior to suture resuspension, with a significantly better reoperation-free and event-free survival. PMID- 10386398 TI - The use of gelatin-resorcin-formalin glue in acute aortic dissection type A. AB - OBJECTIVES: The Gelatin-resorcin-formalin (GRF) glue is widely used in the surgical treatment of dissecting aneurysms. This paper is focused on our experience with the GRF glue in cases, operated for acute aortic dissection type A. METHODS: Between September 1990 and December 1997, 164 patients were operated on for acute aortic dissection type A. In 148 patients GRF was used to reinforce the dissected layers proximal (n = 106) or distal (n = 144) of the grafted aortic segment. An intervention at the aortic valve was necessary in 93 instances. In 111 patients, an open distal anastomosis for replacement of the proximal aortic arch was performed. Thirty-seven additional patients underwent subtotal or total aortic arch replacement. RESULTS: Early postoperative mortality was 26.2% (43/164 patients). Another 16 patients died late postoperatively. Actuarial survival rates are 69.9% at 1 year, 62.5% at 3 years, 59.4% at 5 years and 56.1% at 7 years, post-operatively. Twenty-two reoperations were performed in 20 patients (16.5%). Nine of these patients had developed complications in aortic segments that underwent reconstruction by use of GRF during the primary intervention. Aortic root redissection was found in 7/9 patients intraoperatively, whereas 1/9 patients presented with a rupture near the distal graft to aortic anastomosis. CONCLUSIONS: The introduction of GRF glue has greatly facilitated the reconstruction of dissected aortic wall layers adjacent to the vascular graft. However, the use of the adhesive for aortic root reconstruction in acute aortic dissection type A may bear a significant risk of late postoperative proximal aortic redissection. Complications associated with the GRF glue are likely to be due to the toxic effects of the formalin component. Therefore, care should be taken that the amount of formalin administered to the glue components remains as low as possible. PMID- 10386399 TI - Comparative clinical study between retrograde cerebral perfusion and selective cerebral perfusion in surgery for acute type A aortic dissection. AB - OBJECTIVE: Selection of a brain protection method is a primary concern for aortic arch surgery. We performed a retrospective study to compare the respective advantages and disadvantages of retrograde cerebral perfusion (RCP) and selective cerebral perfusion (SCP) in patients who underwent surgery for acute type A aortic dissection. METHODS: The study reviewed 166 patients who underwent surgery at Nagoya University or its eight branch hospitals between January 1990 and August 1996. There were 91 patients who received SCP and 75 patients who underwent RCP. Results for these two groups were compared. RESULTS: There were no significant differences in age, gender, Marfan syndrome rate, DeBakey classification, or emergency operation rate. Rates of various preoperative complications were similar except for aortic valve regurgitation. Arch replacement was performed more often in SCP than in RCP patients (49% vs. 27%, P = 0.0028). There were no significant differences between groups in cardiac ischemic time or visceral organ ischemic time. However, RCP group showed shorter cardio-pulmonary bypass time (297+/-99 vs. 269+/-112 min, P = 0.013) and lower the lowest core temperature (21.6+/-3.1 degrees C vs. 18.7+/-2.1 degrees C, P = 0.0001). SCP duration was longer than RCP duration (103+/-56 vs. 54+/-24 min, P < 0.0001). Despite these differences, RCP patients were not significantly different from SCP patients with regard to any postoperative complication, neurological dysfunction (16 vs. 19%), or operative mortality (all deaths within the hospitalization; 24 vs. 21%). Regarding neurologic dysfunction, there were six cases of coma, six of motor paralysis, two of paraplegia and one of visual loss among SCP patients, and eight cases of coma, three of motor paralysis, and three of convulsion in the RCP group. The incidence of motor paralysis was higher in the SCP group, while the incidence of coma was higher in the RCP group. CONCLUSIONS: RCP can be performed without clamping or cannulation of the cervical arteries, which is an advantage in reducing the chances of arterial injury or cerebral embolization. RCP is comparable to SCP in terms of clinical outcome. PMID- 10386400 TI - Long-term results after thromboendarterectomy for chronic pulmonary embolism. AB - OBJECTIVE: In patients with chronic thromboembolic pulmonary hypertension, pulmonary vascular resistance (PVR) can be reduced by pulmonary thromboendarterectomy (PTE). In this study, long-term symptomatic and hemodynamic effects were investigated. METHODS: Twenty-two patients (12 female, 10 male, mean age 40 years, preoperative NYHA functional class II/III/IV: n = 1/12/9) were re evaluated 48-72 months (mean 60 months) after surgery. In addition to clinical assessment, radiologic, hemodynamic and echocardiographic investigations were performed. RESULTS: All patients reported a marked improvement of their clinical condition. At follow-up, 11 patients were identified as NYHA class I, 10 as NYHA class II and one patient was in class III. PVR and mean pulmonary artery pressure (mPAP) were significantly reduced (preoperative PVR 800+/-274 dynes/s per cm(-5), follow-up PVR 180+/-28.3 dynes/s per cm(-5); P < 0.001; preoperative mPAP 48.5+/ 7.4 mmHg, follow-up mPAP 27.5+/-4.9 mmHg; P < 0.001). There was also a significant increase in arterial blood oxygen tension (preoperative PaO2 59+/-10 mmHg; follow-up PaO2 84+/-12 mmHg; P < 0.001). Chest roentgenograms and echocardiographic examinations revealed significantly decreased right heart dimensions and a recovery of right heart function. CONCLUSION: In patients with severe chronic thromboembolic pulmonary hypertension, persistent symptomatic and hemodynamic improvements can be achieved by PTE. PMID- 10386401 TI - Lung volume reduction surgery combined with cardiac interventions. AB - OBJECTIVE: Postoperative course and functional outcome were evaluated in patients who underwent lung volume reduction surgery (LVRS) or in combination with valve replacement (VR), percutaneous transluminal coronary angioplasty (PTCA), placement of a stent, or coronary artery bypass grafting (CABG). METHODS: Patients with severe bronchial obstruction and hyperinflation due to pulmonary emphysema were evaluated for lung volume reduction surgery. Cardiac disorders were screened by history and physical examination and assessed by coronary angiography. Nine patients were accepted for LVRS in combination with an intervention for coronary artery disease (CAD). In addition, three patients with valve disease and severe emphysema were accepted for valve replacement (two aortic-, one mitral valve) only in combination with LVRS. Functional results over the first 6 months were analysed. RESULTS: Pulmonary function testing demonstrates a significant improvement in postoperative FEV1 in patients who underwent LVRS combined with an intervention for CAD. This was reflected in reduction of overinflation (residual volume/total lung capacity (RV/TLC)), and improvement in the 12-min walking distance and dyspnea. Median hospital stay was 15 days (10-33). One patient in the CAD group died due to pulmonary edema on day 2 postoperatively. One of the three patients who underwent valve replacement and LVRS died on day 14 postoperatively following intestinal infarction. Both survivors improved in pulmonary function, dyspnea score and exercise capacity. Complications in all 12 patients included pneumothorax (n = 2), hematothorax (n = 1) and urosepsis (n = 1). CONCLUSION: Functional improvement after LVRS in patients with CAD is equal to patients without CAD. Mortality in patients who underwent LVRS after PTCA or CABG was comparable to patients without CAD. LVRS enables valve replacement in selected patients with severe emphysema otherwise inoperable. PMID- 10386402 TI - Excision of pulmonary metastases of osteogenic sarcoma of the limbs. AB - OBJECTIVE: The combination of surgery and chemotherapy improves the prognosis of patients with osteogenic sarcoma of the limbs without detectable metastases at presentation. However, lung metastases are a frequent complication. To evaluate the role of the resection of pulmonary metastases of osteogenic sarcoma of the limbs, we have reviewed our experience with this type of surgery, combined with a multidrug chemotherapy protocol. PATIENTS AND METHODS: From January 89 to December 97, 198 patients operated on for osteogenic sarcomas of the limbs were followed in our centre. Of these, 31 patients (15.7%), with a mean age of 25 years (range 10-54 years), developed lung metastases and had undergone 45 thoracotomies. All patients received chemotherapy, followed by resection of metastatic lesions and additional chemotherapy. The mean time interval between resection of the primary tumour and the diagnosis of lung metastases was 22 months (4-122 months). Eight patients (25.8%) needed more than one (2-4) thoracotomy. The mean time interval between the first and second thoracic surgeries was 9.2 months (2-14 months). RESULTS: There was no operative mortality or major morbidity. During the 45 thoracotomies, five lobectomies and 40 wedge resections were necessary. The mean number of metastases resected per thoracotomy was 3.4 (range 1-10). The degree of necrosis was evaluated by seriated sections for a histologic study. In the end the mean necrotic volume was calculated. A strong correlation was found between the degree of necrosis of the metastases and the need for reoperation for new metastatic lesions, because all the patients who needed more than one operation had less than 80% of necrosis of metastases. The patients were followed for a mean period of 28 months (6-72 months). Ten patients (32.2%) died of related causes at a mean of 19.4 months after thoracic surgery, three of whom had more than one operation. The 3-year survival after metastasectomy was 61%. Patients without pulmonary metastases had a 3-year survival of 79%. CONCLUSIONS: In patients with lung metastases of an osteogenic sarcoma, the combination of chemotherapy and surgery improves the outcome. In our series the mortality was not influenced by the number or thoracotomies required. PMID- 10386403 TI - Is computed tomography guided biopsy still necessary for the diagnosis of adrenal masses in patients with resectable non-small-cell lung cancer? AB - OBJECTIVES: This study was undertaken: (1) to evaluate the usefulness of unenhanced computed tomography (CT), magnetic resonance imaging (MRI) and CT guided biopsy for the characterization of adrenal masses in patients with operable non-small-cell lung cancer (NSCLC) and (2) to evaluate the situations in which CT guided biopsy is absolutely necessary before potentially curative resection of NSCLC. METHODS: Consecutive patients with operable NSCLC underwent unenhanced adrenal CT with density measurements of any adrenal mass over 1 cm in diameter. An adrenal mass was considered as an adenoma when its density was below 10 Hounsfield Units and a metastasis when its density exceeded 10 Hounsfield Units. Then patients underwent MRI, the signal on the T2 weighted images from the enlarged gland was classified adenoma or metastasis in comparison with that from the liver parenchyma. CT guided biopsy was performed after a pheochromocytoma was eliminated. Unenhanced CT attenuation values and signal intensity values on MRI were correlated with histopathologic results. RESULTS: Of the 443 patients, 32 had an adrenal mass consisting of adrenal metastases in 18 cases and adenomas in 14 cases. On CT, 3/14 (21%) of the adenomas were misdiagnosed as metastases (their densities exceeded 10 Hounsfield Units) and 2/18 (11%) of the metastases were misdiagnosed as adenomas (their densities were below 10 Hounsfield Units). On MRI, none of the metastases were misdiagnosed as an adenoma (100% sensitivity) but 7/14 (50%) of the adenomas were misdiagnosed as metastases (signal superior to that of liver). Overall, a diagnostic certainty of metastasis could not be obtained in 25/32 patients (78%). CT guided biopsy with 100% sensitivity and specificity corrected all the inaccurate results of CT and MRI without any morbidity. CONCLUSION: Despite extensive morphological evaluation with unenhanced CT and conventional MRI, CT guided biopsy is necessary for most patients referred to surgery for an operable NSCLC and an adrenal mass. PMID- 10386404 TI - Improvement of pulmonary function after lobectomy for non-small cell lung cancer in emphysematous patients. AB - OBJECTIVE: Pulmonary emphysema is frequently associated with lung cancer and, because of the impaired pulmonary function involved, it may contraindicate surgical treatment. However, improvement of pulmonary function has been observed after surgical resection in patients with advanced emphysema. The aim of this study was to evaluate whether pulmonary emphysema, as assessed by pulmonary function tests and radiological evaluation, can influence postoperative respiratory function after lobectomy for non-small cell lung cancer (NSCLC). METHODS: Respiratory function was evaluated before and after lobectomy for NSCLC. Radiological evaluation of emphysema was performed on chest X-ray and CT scan. Patients that had undergone chemo- or radiotherapy or had segmental or lobar atelectasis were excluded from the study. RESULTS: Thirty-five patients entered the study. A decrease in static lung volumes was observed after surgery. Total lung capacity (TLC) decreased from 6.58+/-0.92 to 5.46+/-0.77 l; functional residual capacity (FRC) from 3.70+/-0.88 to 2.96+/-0.73 1 and residual volume (RV) from 2.93+/-0.78 to 2.2+/-0.53 l. However, in a subgroup of 10 patients (Group 1), dynamic volumes after surgery were unchanged or slightly increased (forced vital capacity (FVC) from 3.23+/-0.65 to 3.3+/-0.68 l; forced expiratory volume in 1 s (FEV1) from 2.14+/-0.51 to 2.25+/-0.54 l), and airway resistances (sRaw) decreased from 15.58+/-5.18 to 11.42+/-5.25 cm H2O/s. Preoperative data showed that these patients had a greater obstruction, with FEV1 changing from 69+/-12.42 to 72.70+/-13.72% of predicted, as compared with a change from 87+/ 12.7 to 72.08+/-13.10% in the other group of 25 patients (Group 2). Correlation analysis reached statistical significance between FEV1% variation (deltaFEV1%) and preoperative FEV1 and FVC% (r = -0.49, P = 0.002 and r = -0.5, P = 0.001, respectively) and between delta (FEV1)% and radiological scores for 3-level CT (r = 0.39, P = 0.04) and the sum of chest X-ray, single and 3-level CT scores (r = 0.49, P = 0.01). CONCLUSIONS: Pulmonary function may remain unchanged or even increase after lobectomy in patients with a pronounced emphysematous component of airway obstruction. The identification of preoperative parameters that identify this group of patients could extend the indications for the treatment of lung cancer in patients with pulmonary emphysema. PMID- 10386406 TI - Tumors of the ribs: experience with 47 cases. AB - OBJECTIVES: To emphasise the existing difficulties in differentiating benign from malignant rib tumours, and especially the problems that a clinical doctor encounters when dealing with a hyperplastic rib. METHODS: Forty-seven patients with rib tumour underwent surgery in a period of 12 years (1984-1996). In 40 cases (85%), the lesion was benign and in seven (15%) was malignant. Twenty-one benign tumours originated from cartilage and bone, seven were inflammatory, six originated from the bone marrow, and minor percentages (2.5-5%) had vascular, neurogenous, degenerative or miscellaneous origin. Three of the malignant tumours were primary chondrosarcomas and two were metastatic from kidney. The rest were metastatic from stomach (adeno-Ca), and skin (melanoma). The mean age in the benign group was 25.2 years and in the primary malignant group was 20.7 years. Related symptoms were pain (47%) and swelling (42.5%). One-third (32%) of the patients were asymptomatic and the lesion was accidentally found during routine chest radiography. All patients were treated surgically with wide excision of the tumour and the diagnosis was established histologically. RESULTS: Resection was complete and curative in all cases without recurrence. CONCLUSIONS: Since the likelihood of malignancy cannot be excluded, all rib tumours should be considered malignant until proven otherwise. Therefore, prompt intervention is necessary and wide and radical initial excision of the involved rib is advocated. PMID- 10386405 TI - Result of induction chemotherapy followed by surgery in patients with stage IIIA N2 NSCLC: importance of pre-treatment mediastinoscopy. AB - OBJECTIVE: Data from the literature indicate that chemotherapy prior to resection may improve the results. However, only few and conflicting data are reported regarding the correlation between downstaging of mediastinal nodes and outcome. The aim of this study was to look at the correlation between downstaging, survival and pre-treatment staging. MATERIAL AND METHODS: Between March 1995 and August 1998, 46 consecutive patients with pathology proven N2 disease were treated with three cycles of vindesine-ifosfamide-platinum (VIP). All patients underwent a rigorously performed cervical mediastinoscopy. Patients with at least partial response (n = 26) were surgically explored. RESULTS: The clinical response rate to chemotherapy was 57% (26 patients). Resection was complete in 23 patients (88.5%). Pneumonectomy was performed in 16 patients. In 11 patients (42.9%) the mediastinal nodes (which were positive at mediastinoscopy) had become negative (downstaging group). The projected 2-year survival of resected patients is 41%. Patients with downstaging of nodes had no better survival compared to patients with no downstaging. Patients with involved subcarinal nodes at mediastinoscopy and patients with involvement of more than one level had a worse survival. CONCLUSION: Surgery in N2-patients responsive to induction chemotherapy resulted in a high complete resectability rate. Findings at pre-treatment mediastinoscopy proved to be the most important prognostic factor. PMID- 10386407 TI - Thymectomy for myasthenia gravis: a 27-year experience. AB - OBJECTIVE: Thymectomy is considered an effective therapeutic option for patients with myasthenia gravis (MG). We reviewed our 27-year experience with surgical treatment of MG with respect to long-term results and factors affecting outcome. METHODS: Between 1970 and 1997, we performed 232 thymectomies for MG. Fifteen patients were lost to follow-up; the remaining 217 form the object of our study. Sixty-two patients (28.4%) had thymoma. Myasthenia was graded according to a modified Osserman classification: 51 patients (23.5%) were in class I, 81(37.3%) in class IIA, 52 (24%) in class IIB, 26 (12%) in class III and seven (3.2%) in class IV. Mean duration of symptoms before the operation was 12+/-10 months. Fifty-eight thymectomies for thymoma were performed through a median sternotomy and four through a clamshell incision. Forty-six thymectomies for non-thymomatous MG were performed through a standard cervicotomy, 101 procedures through a partial upper sternal-splitting incision and eight through a complete median sternotomy. RESULTS: Operative mortality was 0.92% (two patients). After a mean follow-up of 119 months, 71% of all patients improved their clinical status (25% without medications and asymptomatic; 46% with a reduction of medications and/or clinically improved); 39 (18%) have a stable disease with no clinical modifications; 12 (5%) presented a deterioration of their clinical status with worse symptoms, required more medications, or both. Thirteen patients (6%) died because of MG (mean survival 34.3+/-3.6 months). The presence of a thymoma negatively influenced the prognosis. Younger patients showed a more favorable outcome as well as patients with a shorter duration of symptoms before the operation; patients with lower classes of myasthenia showed a higher rate of remission. CONCLUSIONS: Thymectomy is effective in the management of patients with MG at all stages with low morbidity. Patients with thymoma present a less favorable outcome. PMID- 10386408 TI - Oesophagectomy for carcinoma of the oesophagus and oesophagogastric junction. AB - OBJECTIVE: Oesophageal carcinoma has a poor prognosis; surgical resection remains the only chance of cure but is still associated with a significant morbidity and mortality. The aim of this study was to review the results of one surgeon for oesophageal resection for carcinoma of the oesophagus and oesophagogastric junction over a 23 year period. METHODS: Between January 1974 and December 1996, 591 patients (408 males; 183 females; mean age 66 years) underwent an oesophageal resection for carcinoma of the oesophagus or oesophagogastric junction. RESULTS: In hospital mortality was 8.8% (52/591). This has decreased to less than 5% for resections between 1985 and 1996. Non-fatal complications occurred in 21% of patients (123/591). Survival, including in hospital mortality (+/-SEM), was 53.98% (+/-2), 31.77% (+/-2) and 15.3% (+/-2) at 1, 2 and 5 years respectively. CONCLUSION: Early mortality following oesophageal resection has fallen in recent years. Despite considerable experience, long term survival remains disappointingly low. PMID- 10386409 TI - Resection of subaortic stenosis; can a more aggressive approach be justified? AB - OBJECTIVES: Discrete subaortic stenosis causes left ventricular outflow tract (LVOT) obstruction and often produces aortic regurgitation (AR) which alone may precipitate surgical intervention. Conventional resection relieves the obstruction, but the recurrence rate is high, and the AR is little changed as the thick fibrous membrane which extends onto the valve leaflets remains. We studied whether an aggressive surgical approach could reduce both the severity of AR and rate of recurrence of obstruction associated with discrete subaortic stenosis, and whether this aggressive approach could be justified. METHODS: Between June 1992 and April 1996, 37 patients aged 0.5-35 years (median 7.5) underwent resection of a discrete subaortic membrane. Ten underwent re-operation for recurrent obstruction and eight followed previous ventricular septal defect closure. LVOT gradient was measured using the modified Bernoulli equation and AR was graded on a scale of 0-4 (0 = none, 4 = severe). Postoperative assessment was performed early (<7 days) and at mid-term (27.0 months; range 2-59 months). RESULTS: There was significant improvement in AR from mild/moderate to none/trivial (P = 0.019) immediately postoperatively and LVOT gradient from 66.9+/-30.4 to 15.1+/-12.2 mmHg (P < 0.0001). By stepwise logistic regression preoperative gradient correlated significantly with postoperative mild/moderate AR (P = 0.015) and LVOT gradient (P = 0.0036). Preoperative mild/moderate AR also correlated with postoperative mild/moderate AR (P = 0.034). Five patients developed complete heart block, four undergoing reoperation for recurrent obstruction, and one preoperatively had right bundle branch block from previous ventricular septal defect repair. At mid-term follow-up there was no increase in AR or LVOT gradient (14.8+/-12.8 mmHg). Early post-operative AR was the strongest predictor of late mild/moderate AR (P = 0.02). Early post-operative gradient was a weaker predictor (P = 0.04). Pre-operative and early post-operative gradient were significant predictors of late gradient (P = 0.0038; <0.0001, respectively). No patient required reoperation for recurrent obstruction; one underwent late aortic valve replacement for severe AR. CONCLUSIONS: An aggressive surgical approach to discrete subaortic stenosis produces excellent relief of obstruction and frees the valve leaflets, significantly reducing associated AR at early and mid-term follow-up with low morbidity for primary operation. Long-term follow-up is required to confirm whether this early benefit is maintained. PMID- 10386410 TI - Explanted cryopreserved allografts: a morphological and immunohistochemical comparison between arterial allografts and allograft heart valves from infants and adults. AB - OBJECTIVE: Life expectancy of cryopreserved allografts implanted in infants is different from those implanted in adults. A morphological study of explanted allograft heart valves was performed to determine the mechanism of deterioration and to compare cryopreserved arterial and heart valve allografts from adult patients with those explanted from infants. METHOD: Between 1987 and 1996, 209 cryopreserved allografts were implanted: 125 valved conduits or monocusps to reconstruct the right ventricular outflow tract in congenital heart disease, 50 allograft heart valves to treat native aortic and prosthetic aortic valve endocarditis and 34 cryopreserved arterial allografts to replace mycotic aortic aneurysms or infected aortic prosthetic grafts. Two months to 8 years after implantation, 23 heart valve allografts, 11 right-sided and 12 left-sided, and four arterial allografts had to be explanted for reasons such as degeneration, recurrent infection, aneurysm formation or rupture. Besides conventional staining, immunohistochemical detection of cell populations was performed as follows: CD45RO, CD3 and CD43 for T lymphocytes, CD20 for B lymphocytes, CD68 for macrophages, protein S100 for Langerhans-cells, vimentin for fibroblasts, alpha actin for smooth muscle cells and factor VIII for endothelial cells. RESULTS: Explanted cryopreserved allografts were all fibrotic, acellular, non-vital and without endothelial cells. The fibrous tissue was preserved. T lymphocytes, indicating rejection, were found in all right-sided allografts from the paediatric population, but only in 9% of left-sided valves explanted from adults and in one of the four of arterial allografts. Macrophages and Langerhans-cells were found only in right-sided allografts from paediatric patients. CONCLUSION: Right-sided cryopreserved allografts from a paediatric population showed ongoing cellular rejection. By contrast, there was only a weak T-cell mediated rejection to adult heart valve and arterial allografts. Therefore, similar long-term results can be expected in adult arterial and heart valve allografts, whereas longevity of right-sided heart valve allograft in the paediatric age group seems endangered by cellular rejection. PMID- 10386411 TI - Percutaneous mitral commissurotomy versus open mitral commissurotomy: a comparative study. AB - OBJECTIVE: Although many studies in medical literature are comparing percutaneous trans-septal mitral commissurotomy (PTMC) and open mitral commissurotomy (OMC), very few long-term comparative follow-ups are available. METHODS: Between January 1991 and December 1997, 193 patients with isolated mitral stenosis were assigned either to PTMC (111 cases) or to OMC (82 cases). PTMC was performed in all cases with Inoue Ballon, OMC was performed with standard techniques. Categorical values were compared by chi square analysis, whereas continuous data were compared by Mann-Whitney test. Univariate survival and event free analysis (Kaplan-Meier+/-SE and log rank) were performed. Recurrent stenosis was classified any mitral valve area (MVA) less than 1.2 cm2 and whenever post-op. echo showed a loss more than 50% of the initial gain. Data were reported as mean+/-SD. Data concerning late echocardiographic assessment were studied with linear and logistic regression analysis. RESULTS: The two groups were homogenous as far preoperative variables as sex, mean age, MVA, echo score and incidence of left atrial thrombosis were concerned. Mean NYHA was preoperatively higher in OMC (2.79+/-0.58) versus PTMC (2.42+/-0.5) (P = 0.001). There was no hospital mortality in both groups. Incidence of hospital complications was similar (4/ 111 after PTMC and 1/82 after OMC; P = 0.3). Seven year survival: 95.41+/-0.02 (PTMC) and 98.05+/-0.01 (OMC) (P = 0.3) and freedom from late complications did not show statistical differences: Embolism 98.78+/-0.01 in PTMC and 98.78+0.01 in OMC (P = 0.8); Recurrent stenosis 71.89+/-0.13 in PTMC versus 82.89+/-0.08 in OMC (P = 0.2); Reoperation 88.43+/ 0.08 in PTMC versus 96.25+/-0.02 in OMC (P = 0.4). A larger MVA was found in patients undergone to OMC (2.05+/-0.35) versus PTMC (1.81+/-0.33) (P = 0.001). Furthermore mean NYHA was lower in OMC (1.14+/-0.3) versus PTMC (1.39+/-0.7) (P = 0.001). CONCLUSIONS: Both techniques achieve with a low operative risk and low incidence of complications a good palliation of rheumatic mitral stenosis. Incidence of complications in the follow-up is similar. OMC allows a larger mitral valve area, a better functional recovery and a lower incidence of late mitral regurgitation. PMID- 10386412 TI - Ischemic preconditioning improves preservation with cold blood cardioplegia in valve replacement patients. AB - OBJECTIVE: The purpose of this study was to test the hypothesis that ischemic preconditioning improves myocardial protection in valve replacement patients undergoing cold-blood cardioplegic arrest and to study the mechanisms of human myocardial ischemic preconditioning initially. METHODS: Forty patients who required double valve replacement were studied. After the institution of cardiopulmonary bypass, 20 patients were preconditioned with two cycles of 3 min of aortic cross-clamping and 2 min of reperfusion before cardioplegic arrest (group IP). Twenty patients were not preconditioned as controls (group C). All hearts were arrested with 4 degrees C cold-blood cardioplegic solution. During perioperation, the blood samples were collected from coronary sinus and radial artery, which were used to measure calcitonin gene-related peptide (CGRP) and creatine kinase-MB (CK-MB). The right atrial myocardial tissue was collected to measure superoxide dismutase/malondialdehyde (T-SOD/MDA) and to observe myocardial ultrastructure. Hemodynamic date were measured. RESULTS: After reperfusion for 30 min, myocardial MDA was significantly lower in group IP than in group C (2.6+/-0.2 vs. 3.8+/-0.3 nM/mg) and T-SOD was significantly higher in group IP than in group C (13.1+/-12.1 vs. 9.2+/-1.2 IU/mg). Ischemic preconditioning significantly increased the production of myocardial CGRP just after preconditioning (92.0+/-4.1 vs. 52.3+/-4.5 pg/ml) and the begin of reperfusion (95.3+/-3.8 vs. 61.2+/-4.9 pg/ml), and deduced the release of CK-MB at 12 h post-reperfusion (77.5+/-9.2 vs. 136.5+/-8.9 IU/l). Preconditioning also improved cardiac function at 30 min and 12 h after reperfusion (cardiac index 2.8+/-0.3 vs. 2.3+/-0.2 l/min per m2 and 2.9+/-0.1 vs. 2.4+/-0.2 l/min per m2). CONCLUSIONS: Ischemic preconditioning enhance cardioplegic protection in valve replacement patients. The possible protective mechanism was that ischemic preconditioning decreased the production of oxygen free radicals. PMID- 10386413 TI - Bilateral lung transplantation via two sequential anterolateral thoracotomies. AB - OBJECTIVE: Bilateral anterior trans-sternal thoracotomy (clam shell incision) is the standard approach used for bilateral sequential lung transplantation (BLTX). The morbidity of this large incision can be considerable. Two separate sequential anterolateral thoractomies represent a less invasive approach. METHODS: The value of this approach was investigated in a prospective series of 22 consecutive patients who received BLTX between June 1997 and July 1998. Their underlying diseases were COPD (n = 16), cystic fibrosis (n = 4) and other (n = 2). All patients underwent BLTX through two anterolateral thoracotomies, without the use of cardiopulmonary bypass. The anterior mediastinum and the sternum with all the surrounding tissue were left completely intact. Twenty-one patients underwent spirometrical examination during the postoperative in-hospital stay. Follow-up is 7+/-4 months (range: 3 to 15). RESULTS: The only intraoperative complication was severe reperfusion edema of the first transplanted lung seen in one patient at the end of the operation, which required pneumonectomy during the same session. All other operations were uneventful. The difference between the cold ischemic time of the first and second transplanted lung was 83+/-17 min. Median intubation duration, ICU- and in-hospital-stay were 1.5, 5 and 20 days, respectively (ranges: 1 to 96, 2 to 96 and 15 to 96, respectively). One major perioperative complication occurred and was due to gross donor/recipient size mismatch: the patient required lobectomy of the consolidated right upper lobe 11 days after transplantation. In 19 patients (86.4%), this less extensive incision allowed early postoperative mobilization, which resulted in good ventilatory performance, with VC of 53+/-15 and FEV1 of 60+/-20% of the predicted, respectively, at the first spirometry, 3 weeks after the operation. Three months survival was 100%. CONCLUSION: The bilateral sequential anterolateral thoracotomy represents a safe and minimal invasive approach for BLTX compared with the clam shell incision. It minimizes the operative trauma, improves postoperative functional recovery and prevents the potential spread of unilateral complications to the other pleural cavity. PMID- 10386414 TI - Intermediate term results of total lymphoid irradiation for the treatment of non specific graft dysfunction after heart transplantation. AB - BACKGROUND: A proportion of heart transplant recipients develop poor graft function in the absence of cellular infiltrate in endomyocardial biopsies or transplant associated coronary artery disease. The condition has a poor prognosis and its aetiology is poorly understood. We report encouraging intermediate term results with total lymphoid irradiation (TLI) in the management of this condition. METHODS: Eleven adult cardiac transplant recipients who developed severe allograft dysfunction (NYHA class-4) at a median period of 4 months after orthotopic heart transplantation were successfully treated with TLI. Endomyocardial biopsies and coronary angiography were normal in each patient and biventricular failure developed in spite of immunosuppression with Cyclosporin-A, Azathioprine, oral Prednisolone, Cyclophosphamide and intravenous Methylprednisolone therapy. Total lymphoid irradiation was given with standard Mantle and inverted Y-fields over ten treatments to achieve a cumulative dose of 8 Gy. RESULTS: Each patient had a significant improvement in clinical response and in ventricular performance within 2 months of commencing TLI. Nine patients are currently well (four NHYA class-1, five NHYA class-2) at 4-48 (median 26) months following TLI. Two patients died; one from bacterial septicaemia and one as a consequence of chronic renal failure. Three patients developed opportunistic infection which was successfully treated with appropriate antimicrobial agents. An Ebstein-Barr virus associated lymphoproliferative disorder occurred in one patient and was successfully treated by reduction in immunosuppression and high dose Acyclovir. Two patients developed transient bone marrow suppression. CONCLUSION: The intermediate term results of TLI in the management of poor graft function in cardiac transplant recipients with normal endomyocardial biopsies and coronary angiography are encouraging. Although complications of opportunistic infection, bone marrow suppression and lymphoproliferative disorder occurred, treatment was successful in each case. PMID- 10386415 TI - Celsior solution for improvement of currently used clinical standards of lung preservation in an ex vivo rat model. AB - OBJECTIVE: The introduction of Euro-Collins solution with its intracellular electrolyte composition has allowed for clinically accepted pulmonary preservation for up to 7 h of ischemic time. In recent years several alternative solutions have been developed for the improvement of pulmonary preservation. Celsior is an extracellular solution which has significantly reduced the ischemia/reperfusion (IR)-induced pulmonary damage in animal studies. So far, no larger experimental studies exist concerning the influence of Celsior on pulmonary gas exchange following IR. METHODS: In an extracorporeal rat lung model ten lungs, were each preserved with Celsior (CE) and Celsior/ prostacycline (CEPC, 6 mg/100 ml) at 4 degrees C and compared with preservation with low potassium-Euro-Collins solution (LPEC, 40 mmol/l of potassium). After 2 h of ischemia the lungs were re-ventilated and reperfused using a roller-pump. Relative oxygenation capacity (ROC), pulmonary vascular resistance (PVR), peak inspiratory pressure (PIP) and wet/dry ratio were monitored for 50 min. Statistical analysis was performed using ANOVA. RESULTS: ROC was increased in all Celsior preserved organs compared with the EC group (P < 0.032). Though the CEPC group was found to have the lowest PIP and the least amount of lung water as assessed by wet/dry ratio, PVR was highest after 30-50 min. The significantly lowest PVR was determined in the CE group (P < 0.02). CONCLUSIONS: Celsior provides better lung preservation than LPEC solution, as demonstrated by a significantly increased oxygenation ability, a lower PVR and a decreased wet/ dry ratio. In vivo experiments and additional histological analysis are warranted for further evaluation of Celsior in lung preservation. PMID- 10386416 TI - The influence of antibody and complement removal with a Ig-Therasorb column in a xenogeneic working heart model. AB - OBJECTIVE: Organ transplantation is limited by the number of brain-dead human donors. Xenotransplantation could be an alternative to guarantee a constant supply of organs. A major problem of xenotransplantation are xenogeneic natural antibodies (XNAb) directed against species-specific antigens of a discordant donor species (e.g. pig). They trigger the hyperacute xenograft rejection (HXR). Re-usable immunoapheresis (LA)-columns Ig-Therasorb (Therasorb, Baxter) were used to adsorb these XNAb. The effect of immunoapheresis of the perfusing human blood was investigated in ex vivo working pig hearts. METHODS: Hearts of 12 landrace pigs (body weight 14-31 kg) were explanted after inducing cardiac arrest with 4 degrees C Celsior solution. Human blood (500 ml, heparinized) was obtained from healthy volunteers. In group 1 (G1, n = 6), blood as perfusate remained untreated. In group 2 (G2, n = 6), native blood was separated by plasmapheresis into cellular components and plasma. The latter passed through the Ig-Therasorb column for removal of immunoglobulins (so-called immunoadsorption or immunoapheresis). After back-table preparation the hearts were mounted to the working heart model. After 20 min of reperfusion in Langendorff mode, the working heart mode was established. Blood samples were taken isochronously for measurement of: CK(-MB), LDH, ASAT, troponin, immunoglobulins, complement activity, anti-pig antibodies and others. After cessation of the heart, atrial and ventricular tissue samples were taken for histological examinations (light/electron microscopy and immunohistochemistry). RESULTS: Two cycles of immunoapheresis reduced the levels of IgG by 84%, IgM by 83.3% and IgA by 76%. In G2, the antibody immunoadsorption of blood prolonged the duration of the working heart mode significantly to 335+/-37.5 min. In contrast, hearts of group 1 (control) failed after 125+/-31.3 min. Heart rate was significantly different between both groups (G1, 77.3+/-6.1 beats/min; G2, 86.5+/-5.5 beats/min). In G2 cardiac output was 118% and mean coronary flow was 154.6% higher than in G1. CK, LDH and ASAT showed no differences in the two groups. Heart weight increased significantly more in group 1 than in G2. Histological examination indicated specific signs of HXR in G1 after 1.5 h, whereas in G2 only slight unspecific damages were found after 6 h. CONCLUSION: Antibody removal by means of immunoapheresis results in a significantly improved xenogeneic cardiac function. Immunoapheresis may, therefore, become an important adjunct in future pig-to-man clinical xenotransplantation. PMID- 10386417 TI - Angiographic results after minimally invasive coronary bypass grafting using the minimally invasive direct coronary bypass grafting (MIDCAB) approach. AB - OBJECTIVE: The aim of the study was to evaluate the early and mid-term angiographic results after minimally invasive coronary bypass grafting using an 'off-pump' technique via a lateral minithoracotomy. METHODS: In 221 out of 271 patients (81.5%) who underwent minimally invasive direct coronary bypass grafting (MIDCAB) the quality of the internal thoracic artery (ITA)-graft and the anastomosis was evaluated by conventional coronary angiography between the 2nd and 6th postoperative day (POD). A subgroup of 130 patients (47.9%) of the initial cohort were repeatedly controlled by angiography 6 months later. RESULTS: The early postoperatively patency rate of the grafts was (96.8%). Moderate anastomotic stenosis between 50 and 75% was found in 13/221 (5.8%) patients, whereas severe stenosis of more than 75% was seen in 10/221 (4.5%) and occlusion of the graft in 3/221 (1.3%) patients. A stress-ECG was performed in patients with a severe stenosis to provoke ST-segment changes or clinical findings of myocardial ischemia. A positive stress test was found in 4/221 patients (1,8%). Early re-intervention was required in 7/221 (3.1%) patients. After 6 months, angiographic follow-up revealed a patency rate of (95.4%). Of 130 patients 5 (3.8%) presented with moderate anastomotic stenosis, whereas 3/130 (2.0%) patients showed a severe stenosis with one patient (0.7%) having myocardial ischemia during stress test. Occlusion of the graft was seen in 3/130 patients (2.3%). During follow-up, 4/130 (3.0%) patients underwent re-intervention. A comparison between early postoperative and 6-months angiogram revealed a decrease or a disappearance of the severity of the stenosis in 4/15 patients (26.6%). CONCLUSION: Since stenosis of the anastomosis may occur after minimally invasive, beating heart coronary bypass grafting, postoperative angiography should be performed to provide quality control and to guide appropriate further treatment. The latter is necessary if the stenosis is accompanied by reduced run-off and evidence of myocardial ischemia during stress test. An improvement of early stenosis at the anastomosis may be expected in more than 25%. PMID- 10386418 TI - Beating versus arrested heart revascularization: evaluation of myocardial function in a prospective randomized study. AB - OBJECTIVE: Conventional coronary artery bypass grafting (CABG) is both safe and effective. Nevertheless, the use of cardiopulmonary bypass (CPB) and cardioplegic arrest are associated with several adverse effects. Over the last 2 years there has been a revival of interest in performing CABG on the beating heart. In this prospective randomized study we evaluated the efficacy and safety of on and off pump coronary revascularization on myocardial function. METHODS: Eighty patients (65 males, mean age 61+/-9.7 years) undergoing first time CABG were prospectively randomized to: (i) conventional revascularization with CPB at normothermia and cardioplegic arrest with intermittent warm blood cardioplegia (on pump) or (ii) beating heart revascularization (off pump). Troponin I (Tn I) release was serially measured as a specific marker of myocardial damage. Haemodynamic measurements as well as inotropic requirement, incidence of arrhythmia and postoperative myocardial infarction were also recorded. RESULTS: There were no significant differences between the two groups in terms of age, sex, extent of disease, left ventricular function and number of grafts. There were no deaths or intraoperative myocardial infarctions in either group. Tn I release was constantly lower in the off pump group and this was significant at 1, 4, 12 and 24 h postoperatively. Furthermore, in this group there was a significantly reduced incidence of arrhythmias. Inotropic requirements were less in the off pump group but this did not reach statistical significance. CONCLUSION: These results suggest that off pump coronary revascularization is a safe and effective strategy for myocardial revascularization. Myocardial injury as assessed by Tn I release is also reduced when compared with conventional coronary revascularization with CPB and cardioplegic arrest. PMID- 10386419 TI - Early and long term results of coronary artery bypass grafts in patients with dialysis dependent renal failure. AB - OBJECTIVE: Coronary artery disease is the main cause of mortality and morbidity in patients on renal therapy replacement. The aim of this study was to define peri-operative risk and long term results of coronary artery bypass grafts (CABG) in dialysis patients. METHODS: this retrospective study included 82 patients in chronic dialysis who underwent CABG between 1978 and 1997. The mean age was 61+/ 10 years (range 28-81 years), 84% of the patients were male and the average duration of dialysis was 57 months (range 1-148 months). Combined procedures were carotid endarterectomy in one case, left ventricular aneurysm resection in one and valvular replacement in 10 (nine aortic and one mitral replacements). The operation was elective in 42 patients (51 %) and urgent in the others. Previous myocardial infarction was found in 37 patients (45%) and left ventricular ejection fraction (LVEF) at less than 45% in 15 patients (18%); 23 patients (28%) were in NYHA class III or IV and regarding angina functional status, 77% in CCS class 3 or 4. Follow-up was complete. Statistical analysis included 30 and pre and peri-operative data. Statistical analysis used Chi-square analysis or Fisher's exact test, and the Mann-Whitney test when appropriate. The estimated probability of survival, including postoperative mortality, was calculated by the method of Kaplan-Meyer, and the Log-Rank test used to compare the results. RESULTS: the hospital mortality was 14.6 % (n = 12). Ischemic time and ECC time were significantly lengthened in dead patients (P = 0.01). Moreover, use of internal mammary artery was directly related to lower hospital mortality (P = 0.02). For previous myocardial infarction, LVEF at less than 45%, diabetes and combined procedure, a P-value of < or = 0.1 was calculated. The follow-up ranged from 1 to 140 months (mean 36 months). There were 39 late deaths. The survival rates (included hospital mortality) were 71+/-5%, 56+/-6% and 39+/-6% at 1, 3 and 5 years, respectively. All surviving patients improved their functional status and had symptomatic relief. Statistical analysis showed significant difference in favor of long term survival for patients younger than 60 years, LVEF > 45% and NYHA class I or II. CONCLUSION: these data confirm that CABG in patients with renal replacement therapy is associated with an high operative and long term mortality. However it allows an improvement of functional status, and so, let possible duration of dialysis. It may be expected that more active prevention and detection of coronary disease might improve these results. PMID- 10386420 TI - Coronary artery bypass grafting in patients with systemic lupus erythematosus. AB - OBJECTIVE: Few reports exist on the results of coronary artery bypass grafting (CABG) in patients with systemic lupus erythematosus (SLE). METHODS: We retrospectively reviewed eight CABG in seven SLE patients. In early and late postoperative angiography, all grafts were evaluated for occlusion, development of string sign, or presence of significant stenosis. The early and late results were compared. The pathological studies were performed on the segments of the internal thoracic artery (ITA) and saphenous vein collected from each patient. Atherosclerosis of the ITA was analyzed using the subjective evaluation proposed by Kay et al. (Kay HR, Korns ME, Flemma RJ, Tector AJ, Lepley D. Atherosclerosis of the internal mammary artery. Ann Thorac Surg 21;1976:504-507) scale 0-4 (0 = normal, 1 = minimal disease, 2 = less than 25% luminal narrowing, 3 = 25-50% narrowing, and 4 = greater than 50% narrowing). RESULTS: The patients consisted of three men and four women with a mean age of 59.8 years. Co-morbid diseases were frequent and there were three patients (37.5%) with renal failure (two dialysis patients, one with renal dysfunction) and two patients with severe atherosclerosis of the aorta. The ITA was used in four patients. Saphenous vein graft was used in seven patients. Concomitant procedures included aortic valve replacement and mitral annuloplasty, mitral valvuloplasty and tricuspid annuloplasty, mitral valve replacement and tricuspid annuloplasty (TAP). There was one hospital death (12.5%). Early patency rates were 87.5% (21/24). No other atherosclerotic changes or stenosis suggesting vasculitis were noted. In pathological studies, there was no significant atherosclerosis in the six ITA specimens from four patients, although three patients had degree two atherosclerosis. No vasculitis was found in ITA or saphenous vein grafts. During the mean follow-up period of 35.3 months (range, 5-91 months), there was one non cardiac late death. Late restudy (in three patients, 12, 57 and 64 months later respectively) revealed no deterioration in either ITA or vein grafts. Overall prognosis after the operation in SLE patients appears to be good. No other cardiac events were observed, and patients demonstrated marked clinical improvement. CONCLUSIONS: CABG in SLE patients can be performed with acceptable morbidity and mortality. Our data so far reveals no evidence to preclude the use of ITA and vein grafts in SLE patients. PMID- 10386421 TI - Transmyocardial laser revascularization in the acute ischaemic heart: no improvement of acute myocardial perfusion or prevention of myocardial infarction. AB - OBJECTIVE: Transmyocardial laser revascularization (TMLR) has been used to provide enhanced myocardial perfusion in patients not suitable for coronary revascularization or angioplasty. This study investigates the acute changes in myocardial perfusion after TMLR with a Holmium:Yttrium-Aluminium-Garnet (YAG) laser with a thermal imaging camera in a model of acute ischaemia, and confirms its midterm effects by post-mortem investigation of magnetic resonance imaging and histopathological examination. METHODS: Acute myocardial ischaemia was induced by occlusion of the dominant diagonal branch in ten sheep. Perfusion measurements were undertaken first in the unaffected myocardium, then after temporary occlusion of the coronary to obtain a control measurement for ischaemic myocardium. Myocardial perfusion was then evaluated during reperfusion after release of coronary occlusion. Then the coronary was permanently occluded and 20.5+/-2 channels were drilled with the Holmium:YAG laser and perfusion was measured again. The other four sheep served as control with untreated ischaemia. All animals were sacrificed after 28 days following administration of gadolinium i.v. to serve as contrast medium for magnetic resonance tomography. The hearts were subjected to magnetic resonance tomography and histopathological examination. RESULTS: Intraoperative perfusion measurements revealed a decreased perfusion after temporary occlusion and an increased perfusion in reperfused myocardium. After TMLR, no improvement of myocardial perfusion above the ischaemic level could be shown. Magnetic resonance images could neither confirm patent laser channels nor viable myocardium within ischaemic areas. On histology no patent endocardial laser channel could be detected. The transmural features were myocardial infarct with scar tissue. CONCLUSIONS: In the presented sheep model with acute ischaemia, TMLR with a Holmium:YAG laser did not provide acute improvement of myocardial perfusion as assessed by a thermal imaging camera. This would suggest no direct contribution of newly created laser channels to myocardial perfusion. As chronic effects are concerned, no perfused laser channels could be identified by later magnetic resonance imaging or histology. PMID- 10386422 TI - Analysis of the new indirect revascularization method by determining objective parameters of clinical chemistry, histo-chemistry and histology. AB - OBJECTIVE: This experimental study was initiated to determine whether transmyocardial laser revascularization (TMLR) after acute myocardial ischemia may improve clinical chemistry and diminish the amount of necrosis. In addition, the influence of TMLR on healthy myocardium was analyzed. METHODS: The prolonged short-term effectiveness of TMLR was evaluated in 44 open-chest anesthetized pigs with (n = 21) or without (n = 23) the setting of acute myocardial ischemia (observation period 6 h): seven pigs served as controls (thoracotomy only). An additional seven pigs had left anterior descending artery (LAD) occlusion only (ischemia group). A subsequent 14 pigs were treated by TMLR (CO2) prior to LAD occlusion: Seven pigs received one laser channel/cm2 (group 1) and in seven pigs two channels/cm2 in the LAD territory (group 2) were performed. In addition, 16 pigs underwent TMLR without ischemia: Eight pigs received one channel/cm2 (group 3) and eight pigs two channels/cm2 (group 4). Clinical chemistry, histo-chemical assessment and histology were performed. RESULTS: TMLR limits the expansion of the myocardial infarction zone: laser group 2 indicated a significant smaller area of necrosis in the area at risk (ischemic group (31%) vs. laser group 1 (19%), P = ns; laser group 2 (7%) vs. ischemic group, P < 0.01; laser group 1 vs. 2, P < 0.01). The amount of the area of necrosis and ischemia of laser groups 3 and 4 compared with control did not differ significantly (P = ns). Preventive creation of microchannels before ischemia did not diminish ischemic parameters (P = ns). The myocardial water content-measurements (MWC) in the ischemia, laser 1 and 2 groups did not show any difference at the end of the experiment, except higher values of laser group 2 (P < 0.05). Laser groups 3 and 4 revealed significantly higher MWC values compared with control (P < 0.001). CONCLUSIONS: This prolonged acute study demonstrates that CO2-TMLR significantly reduces the amount of necrosis in the area at risk, but does not reduce cardiac ischemic markers. In healthy myocardium, TMLR significantly increases myocardial water content and ischemic parameters and induces small ischemic and very small necrotic areas surrounding open laser channels. Generally, the elevated cardiac enzymes and proteins are mainly attributed to the expected rise caused by vaporization of myocardial tissue in all laser groups. PMID- 10386423 TI - The effect of 'renal-dose' dopamine on renal tubular function following cardiac surgery: assessed by measuring retinol binding protein (RBP). AB - OBJECTIVE: Acute renal failure complicating open heart surgery is not uncommon. Dopamine infusion (2.5-4.0 microg/kg per min) has often been advocated for prophylactic 'renal protection' in this setting despite little objective evidence of real benefit. We aimed to investigate whether dopamine offers any 'renal protection' in patients with normal heart and kidney functions undergoing routine coronary artery bypass grafting (CABG). Urinary excretion of retinol-binding protein (RBP), previously validated as a sensitive and accurate marker of early renal tubular injury, was used to assess the renal effects of dopamine during the first postoperative week. METHODS: Forty consecutive patients from the elective waiting list were prospectively randomized into two equal groups: those in Group A received dopamine infusion at 'renal dose' (2.5-4.0 microg/kg per min) starting from induction of anaesthesia for 48 h, whereas those in Group B served as untreated controls. Daily measurements were made of weight-adjusted urine output (ml/kg), fluid balance (input/output), serum creatinine, blood urea and urinary RBP. Statistical comparisons were made using Mann-Whitney U-test. RESULTS: The two groups matched in terms of age, time and temperature on cardiopulmonary bypass, number of grafts performed and perioperative haemodynamic status. No differences were detected in the weight-adjusted urine output, fluid balance, serum creatinine and blood urea between the groups. Control subjects (Group B) showed an increase in urinary RBP during the first and second postoperative days (323+/-4 microg/ mmolCr and 50+/-3 microg/mmolCr; mean+/-SD). However, patients treated with dopamine (Group A) demonstrated much greater urinary excretion of RBP over the same period (1257+/-15 microg/mmolCr and 449+/-21 microg/mmolCr; P = 0.0006 and 0.03) than those in Group B. CONCLUSIONS: Dopamine given at 'renal dose' appears to offer no renal protection in patients with normal heart and kidney functions undergoing elective coronary surgery. On the contrary, it exacerbates the severity of renal tubular injury during the early postoperative period. Based on these findings we do not recommend the use of dopamine for routine renal prophylaxis in this group of patients. PMID- 10386424 TI - Desmoid fibromatosis of the shoulder and of the upper chest wall following a clavicular fracture. AB - A desmoid tumor of the shoulder girdle infiltrating the upper chest wall and weighing 1500 g was almost completely removed in an 18-year-old man, 27 months after a bifocal fracture of the clavicule. Thirteen years later, the patient was free of recurrence. The interval time between trauma and diagnosis, as the particular characteristics of aggressive fibromatosis, strongly support a major causal role of the clavicular fracture in the occurrence of this tumor. PMID- 10386425 TI - Metachronous pulmonary and oesophageal neoplasia. AB - Primary carcinomas of the lung and oesophagus are common, surgical resection offers the only hope of long-term survival with both conditions. We present the unusual case of a patient who underwent transhiatal oesophagectomy for an adenocarcinoma carcinoma of the oesophagus, 5 years after left pneumonectomy for small cell carcinoma of the lung. PMID- 10386426 TI - Coronary artery bypass graft after esophagogastrectomy. AB - A 71-year-old male with a history of retrosternal gastric bypass, after a resected esophageal carcinoma, developed angina pectoris due to stenosis of the left main trunk and the left anterior descending artery. The patient was treated with off-pump beating-heart coronary artery bypass approached via left thoracotomy. Two free conduits arising from the left internal mammary artery were utilized for this particular case, since the aortocoronary bypass was impossible due to the severely calcified aorta. Postoperative angiography confirmed good coronary flow and the patient has been symptom free for 6 months. PMID- 10386427 TI - This wormy world. 1947. PMID- 10386428 TI - How much human helminthiasis is there in the world? PMID- 10386429 TI - Strongyloides ratti: migration study of third-stage larvae in rats by whole-body autoradiography after 35S-methionine labeling. AB - In order to clarify the migration pathway of Strongyloides ratti, Wistar rats were given 5,000 35S-labeled infective larvae subcutaneously and killed at 10, 15, 20, 25, 30, 40, and 50 hr postinfection. Prior to inoculation, the specific radioactivity level was assessed in the labeled larvae using a scintillation counter. The frozen rat specimens were sectioned at 50 microm, and the sections were freeze-dried and mounted on X-ray film in darkness. The labeled larvae appeared as dark spots on the film after 14 days of exposure. The infected larvae remained at the inoculated site (lower abdomen) until 10 hr after infection. Some larvae were found in the head portion, whereas others existed sporadically in the skin, liver, and lungs at 15 hr. After 20 and 25 hr, the majority of larvae had accumulated in the head portion. Many larvae appeared in the cranial and nasal cavities; however, no larvae were found in any other organs or tissues. At 30 hr, most larvae had begun to accumulate in the ethmoid region again. At 40 and 50 hr, some larvae were recognized in the ethmoid region, and most had already reached the small intestine. This suggests that the larvae directly move to the nasofrontal portion through the subcutis, rather than migrating to the head through either the viscera, ascending vessels, or the foramen occipital magnum. PMID- 10386430 TI - Histogenesis in the metacestode of Echinococcus vogeli and mechanism of pathogenesis in polycystic hydatid disease. AB - Histogenesis of the metacestode of Echinococcus vogeli was traced mainly in rodents inoculated intraperitoneally with finely minced infective vesicles. The fragments aggregated in the peritoneal cavity and coalesced, forming structures (plaques) from which primary vesicles arose. From primordia in their germinal tissue, exogenous vesicles developed, enlarged, and migrated outward to the surface of the laminated membrane, where they remained attached and proliferated. Each unit of vesicles so formed retained discrete identity and, within 6-8 mo, acquired an adventitia; thereafter, exogenous multiplication ceased and endogenous proliferation supervened. Large numbers of daughter cysts arose in the germinal tissue lining chambers within the units; endogenous proliferation also finally ceased, and the daughter cysts produced brood capsules containing protoscoleces. Primordia of exogenous vesicles were not observed in the walls of daughter cysts. Production of protoscoleces involved 3 processes: they developed in typical brood capsules, singly in minute brood capsules, or directly from germinal tissue. Exogenous proliferation is not characteristic in the natural intermediate host of E. vogeli, the paca. Evidently in primates, the initial proliferation in the liver is followed by extension of the metacestode into the peritoneal cavity and eventual invasion of abdominal and thoracic organs. Exogenous proliferation by a process unique to E. vogeli accounts for the clinical course of polycystic hydatid disease. PMID- 10386431 TI - Trematode accumulation by the estuarine gastropod Ilyanassa obsoleta. AB - The accumulation of larval trematodes by Ilyanassa obsoleta (Gastropoda) at 2 estuarine sites in Delaware was studied. Initial infection status of snails was assayed by looking for shed cercariae. Native snails (most already infected) were deployed at sites A and B, and sentinel snails (putatively uninfected) were deployed at site B. All were individually marked and, if found, reassessed for infection after being free 1-3 summers. Himasthla quissetensis, Lepocreadium setiferoides, Zoogonus rubellus, Austrobilharzia variglandis, and Gynaecotyla adunca infections were observed in recovered snails. At site A, in 1993, 62 natives were recovered. Among the 26 initially testing uninfected, 15 had infections at recovery. Of 36 that released cercariae, 26 had the infection initially indicated. Of sentinels released at site B in 1996, 3 of 16 had infections when recollected. One probably was infected before transplant; at least 2 were infected at site B during 1996-1997. Among site B natives, 26 were later examined by dissection in 1996; 22 had the infection status initially revealed. Some site B natives (n = 35) escaped recapture in 1996 but were found in 1997, or 1998, or both, and were reexamined, most by cercarial release. The same cercariae were produced by 30. Among natives (both sites, n = 123), 27.6% exhibited some difference in infection status compared to the initial assay. This probably overestimates changes. Some differences were real but most can be discounted as cases where initial infection status was misrepresented. PMID- 10386432 TI - Investigation of venereal, transplacental, and contact transmission of the Lyme disease spirochete, Borrelia burgdorferi, in Syrian hamsters. AB - A hamster was inoculated with the SI-1 strain of Borrelia burgdorferi and subsequently served as a host to larval Ixodes scapularis Say. Approximately 68% of the nymphs resulting from the fed larvae were infected. Nymphs from this group were fed on uninfected hamsters, and 3 of 4 males and 6 of 6 females became infected. The infected hamsters were allowed to mate with uninfected partners to test for venereal transmission. Six infected females were mated with 6 uninfected males, whereas 3 infected males were mated with 6 uninfected females. None of the uninfected hamsters became infected after mating. Two protocols were used to determine if transplacental transmission of B. burgdorferi occurred. One group included 6 nonpregnant infected females that were subsequently mated and became pregnant. Three of the females were allowed to carry to full term, whereas the other 3 were killed prior to parturition. All fetuses and offspring were negative for B. burgdorferi based on cultures and monoclonal antibody assays. Another group of 6 females was infected via tick bite after becoming pregnant; those females were allowed to carry fetuses to birth and all were negative. Attempts at contact transmission of B. burgdorferi from 2 infected females to 2 uninfected male and 2 uninfected female hamsters and from 2 infected males to 2 uninfected male and uninfected female hamsters via urine or feces failed. PMID- 10386433 TI - Differences in the second internal transcribed spacer (ITS-2) of eight species of gastrointestinal nematodes of ruminants. AB - Genetic differences in the nucleotide sequence of the second internal transcribed spacers (ITS-2) among Trichostrongylus axei, Trichostrongylus colubriformis, Ostertagia ostertagi, Cooperia oncophora, Cooperia punctata, Nematodirus helvetianus, Nematodirus filicollis, and Haemonchus contortus are described. The ITS-2 sequences of the 8 species ranged between 230 and 241 base pairs in length. Sequence similarities between the different genera varied between 60% and 80%. Identities between the different species within a genus varied between 99% for C. oncophora and C. punctata, 95% for T. axei and T. colubriformis, and 89% for N. helvetianus and N. filicollis. The ITS-2 sequences proved to be useful for species differentiation. Except for the species of Cooperia (2.07% intraspecific variations for C. oncophora and 0.83% for C. punctata) the degree of intraspecific variations (N. filicollis 0.85%, T. colubriformis 1.26%, T. axei 1.27%, H. contortus 2.60%, O. ostertagi, and N. helvetianus no variation) was markedly lower than the interspecific variations allowing a reliable differentiation within the ITS-2 region between single species. PMID- 10386434 TI - Genetic diversity and recruitment pattern of Schistosoma mansoni in a Biomphalaria glabrata snail population: a field study using random-amplified polymorphic DNA markers. AB - Random-amplified polymorphic DNA markers have been used to assess the amount and the distribution of the genetic diversity of Schistosoma mansoni within a natural population of Biomphalaria glabrata at a transmission site of the murine schistosomiasis focus of Guadeloupe. Despite high infection rate and heavy schistosome load within the definitive hosts (Ratus rattus), prevalences within intermediate snails ranged from 0.2 to 4.8%. Whatever the transmission season may be (rainy vs. dry), most of the infected snails were spatially aggregated and 88.4% of them harbored a single parasite genotype indicative of a monomiracidial infection; 4.7% had dual sex infections and a parasite intensity not exceeding 3 miracidia per snail. A substantial resistance level toward the parasite and recruitment regulatory process within snails may explain in part the observed low parasite prevalences and intensities. Considering such a distribution pattern of larval S. mansoni genetic diversity among B. glabrata, mobility of the definitive hosts, or rapid turnover of infected snails, or both, are required to maintain genetic heterogeneity within adult schistosome populations. PMID- 10386435 TI - Cold stress-induced modulation of cell immunity during acute Toxoplasma gondii infection in mice. AB - Infection with Toxoplasma gondii in the acute phase results in nonspecific suppression of immunologic function in mice and humans. The present study examined the effects of a physical stressor, i.e., cold stress (CS), on macrophage function (nitrite production, parasite survival) and splenic blastogenesis in the acute phase of murine T. gondii infection. In our stress paradigm, female BALB/c mice were placed in cold water (1 +/- 0.5 C), 5 min each day for 8 days. Nitrite production and parasite survival were measured in cultured peritoneal macrophages obtained from mice subjected to CS after in vivo activation with interferon-gamma/lipopolysaccharide (CS + ACT), and in vitro infection with T. gondii tachyzoites. Peritoneal macrophages from CS + ACT mice showed decreased nitrite production compared to control but activated cells (ACT). Spleen cell proliferation to in vitro stimulation with the mitogens concanavalin A (Con A) and anti-CD3, and Toxoplasma lysate antigen (TLA) was measured in splenocytes obtained from BALB/c mice during the acute phase of infection with T. gondii. Mice subjected to CS and infection (CS + INF) had maximum splenocyte proliferation on days 8 and 15 followed by a subsequent decline on day 28 postinoculation (PI). In contrast, infected mice not subjected to stress (INF) showed decreased splenocyte proliferation on days 8 and 15 followed by an increase on day 28 PI. The rate of mortality was decreased in the CS + INF compared to the INF group during acute infection. These results suggest that CS may alter the pathogenesis of T. gondii infection by modulating acute phase responses, provoking a state of transient disequilibrium between the host and parasite. PMID- 10386436 TI - Characterization of glutathione S-transferase of Taenia solium. AB - A Taenia solium glutathione-S-transferase fraction (SGSTF) was isolated from a metacestode crude extract by affinity chromatography on reduced glutathione (GSH) sepharose. The purified fraction displayed a specific glutathione S-transferase (GST) activity of 2.8 micromol/min/mg and glutathione peroxidase selenium independent activity of 0.22 micromol/min/mg. Enzymatic characterization of the fraction suggested that the activity was closer to the mammalian mu-class GSTs. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, gel filtration, and enzyme activity analysis showed that the fraction was composed of a major band of Mr = 26 kd and that the active enzyme was dimeric. Immunohistochemical studies using specific antibodies against the major 26-kd band of the SGSTF indicated that GST protein was present in the tegument, parenchyma, protonephridial, and tegumentary cytons of the T. solium metacestode. Antibodies generated against the SGSTF tested in western blot showed cross-reactivity against GSTs purified from Taenia saginata, T. taeniaeformis, and T. crassiceps, but did not react with GSTs from Schistosoma mansoni, or mice, rabbit, and pig liver tissue. Furthermore, immunization of mice with SGSTF reduced the metacestode burden up to 74.2%. Our findings argue in favor of GST having an important role in the survival of T. solium in its hosts. PMID- 10386437 TI - Plagiorchis elegans (Digenea: Plagiorchiidae) infections in Stagnicola elodes (Pulmonata: Lymnaeidae): host susceptibility, growth, reproduction, mortality, and cercarial production. AB - Eggs of Plagiorchis elegans were readily ingested by Stagnicola elodes of all ages, but were more infective to immature than mature snails. Infection enhanced the growth of the host in a dose-dependent manner. The number of cercariae released by immature snails increased with the age of the snail host; mature snails yielded fewer cercariae. Heavily infected snails tended to die prematurely, thereby reducing their total production of cercariae to levels below those of more lightly infected individuals. Even light infections castrated the snail host. Snails that acquired the infection as juveniles never produced eggs. Actively reproducing snails ceased egg laying within days of infection and never recovered. All parasite effects on the growth and reproduction of the snail host first manifested themselves during the early stages of infection, long before the development of daughter sporocysts and cercariae, and are therefore attributable to the effects of mother sporocysts. The study provides insight into how this natural enemy of mosquito larvae may be established in natural snail populations by means of strategically timed introductions of parasite eggs, with a goal of maximizing cercarial production for the biological control of sympatric mosquito larvae. PMID- 10386438 TI - The oral route as a potential way of transmission of Schistosoma bovis in goats. AB - The infectivity of Schistosoma bovis cercariae administered orally was evaluated in Sahelian goats. Compared to the percutaneous route, a single massive oral dose resulted in a worm burden and in fecal egg excretion reduced by one-half. Surprisingly, tissue egg counts were increased by more than 4-fold. Fecundity of individual female schistosomes was, therefore, markedly increased. When infective doses were administered weekly for 20 wk, both worm and egg burdens were doubled without modification of the individual worm pair fecundity. Repeated oral infections seem to have induced an acquired tolerance toward parasite antigens. These results confirm the epidemiologic relevance of the oral route in a host species inclined to become infected through drinking water rather than percutaneous exposures. PMID- 10386439 TI - Susceptibility and carrier status of impala, sable, and tsessebe for Cowdria ruminantium infection (heartwater). AB - Three species of wild African ruminants, impala (Aepyceros melampus), sable (Hippotragus equinus), and tsessebe (Damaliscus lunatus), were experimentally inoculated with in vitro culture-derived Cowdria ruminantium organisms, the tick borne causative agent of heartwater in domestic ruminants, to determine their susceptibility to infection. No clinical disease was observed in any of the ruminants. However, C. ruminantium was detected in the sable by the transmission of heartwater to susceptible sheep, through the tick vector Amblyomma hebraeum, at 10 and 37 days postinfection (PI). Attempts to detect infection in the impala and tsessebe by tick transmission at 54 days PI failed. The impala and tsessebe were reinoculated with C. ruminantium organisms at 146 days after the first inoculation; however, a tick transmission attempt at 66 days after the reinoculation also failed. Seroconversion, as detected by immunoblotting, was demonstrated in the sable and the tsessebe but not in the impala. The results demonstrate that sable can be carriers of C. ruminantium. The susceptibility of tsessebe and impala, however, remains undetermined. PMID- 10386440 TI - Bombesin-like neuropeptides in nematodes. AB - This paper reports evidence of members of the bombesin-like peptide family in a number of nematodes, including Caenorhabditis, Panagrellus, Dirofilaria, Onchocerca, Brugia, Haemonchus, Ostertagia, Toxocara, and Ascaris. One of these (Ostertagia) secretes the bombesin-like material into its environment. Specific binding of gastrin-releasing peptide to the hypodermis consistent with the presence of receptors was demonstrated. These data suggest that this class of peptides may play an important role in nematode hypodermal physiology. PMID- 10386441 TI - Endothelial cell changes are associated with pulmonary edema and respiratory distress in mice infected with the WA1 human Babesia parasite. AB - A C3H/HeN mouse model was established to study the pathogenesis of the human babesial parasites, WA1 and Babesia microti. To evaluate the course of parasitemia and the associated lesions, mice were inoculated intraperitoneally with either WA1-infected, B. microti-infected, or uninfected hamster red blood cells. WA1-infected mice developed dyspnea and moderate parasitemias, after which death occurred. Babesia microti-infected mice experienced low parasitemias with no apparent morbidity or mortality. WA1-infected mice were thrombocytopenic but not anemic. Hemograms for B. microti-infected mice were similar to controls. Postmortem examination of WA1-infected mice revealed prominent lesions in the lungs, including pulmonary edema and intravascular margination of leukocytes. No pulmonary changes were detected in B. microti-infected mice. Blood gas measurements of WA1-infected mice showed reduced oxygen saturation and pH, and increased carbonic acid compared to controls, indicating hypoxia and respiratory acidosis. Ultrastructure studies of WA1-infected lungs showed hypertrophied endothelial cells containing transcellular channels associated with protein-rich intra-alveolar fluid. Endothelial cell activation was demonstrated by an upregulation of intercellular adhesion molecule-1 in the lungs of WA1-infected mice. The results suggest that recruitment of inflammatory cells to the lungs in WA1-infected mice induces endothelial cell alterations, leading to pulmonary edema and acute respiratory failure. PMID- 10386442 TI - Scyphophyllidium uruguayense n. sp. (Eucestoda: Tetraphyllidea) in Mustelus mento (Cope, 1877) (Chondrichthyes: Carcharhiniformes: Triakidae) from La Paloma, Uruguay. AB - A new species of Scyphophyllidium inhabits Mustelus mento near La Paloma, Uruguay. It resembles Scyphophyllidium giganteum from the Atlantic Ocean and specimens identified as S. giganteum from California by having anapolytic strobilae 155-258 mm long, 250-300 craspedote proglottids, scoleces 1.2-1.4 mm wide, necks 34-41 mm long, immature and mature proglottids wider than long, gravid proglottids wider than long to longer than wide, genital pores averaging 28% of proglottid length from the anterior end, relatively flat ovaries with digitiform lobes reaching the lateralmost extent of the testicular field, vitellaria in 2 fields converging toward the proglottid midline, straight and short cirrus sacs, and postvaginal vas deferens. The bothridia of the new species have accessory bothridial suckers that are smaller than those of California specimens; European specimens reportedly lack accessory bothridial suckers. The new species possesses a uterine duct that joins the uterus at the level of the genital atrium and ventral osmoregulatory ducts medial rather than lateral to the dorsal ducts, an arrangement described for Californian but not European specimens. It differs from both European and Californian specimens by having longer cirri, more testes per proglottid, prominent scales covering the neck, and vaginae and uterine ducts coiled immediately preovarially. Pithophorus, Marsupiobothrium, and Scyphophyllidium may form a clade. PMID- 10386443 TI - Six new Eimeria species from vespertilionid bats of North America. AB - Twenty species of bats (Molossidae, Vespertilionidae) were collected from California, New Mexico, Oregon, South Carolina, Utah, and Baja California Norte (Mexico), and 29 of 404 (7%) animals, including Antrozous pallidus, Eptesicus fuscus, Myotis auriculus, Myotis californicus, Myotis ciliolabrum, Myotis evotis, Myotis lucifugus, Myotis thysanodes, Myotis vivesi, Myotis volans, Myotis yumanensis, and Nycticeius humeralis were infected with Eimeria spp., which represent 6 new species. Sporulated oocysts of a new species from A. pallidus are subspheroidal, 24.8 x 21.6 (22-27 x 19-24) microm with a polar granule and a large globular residuum. The oocyst wall is sculptured, with 2 layers, approximately 1.5 thick. Ovoidal sporocysts are 11.5 x 7.8 (9-13 x 7-10) microm, with Stieda body and residuum of many large granules. Sporulated oocysts of a new species from M. californicus are subspheroidal, 20.7 x 18.2 (19-23 x 16-20) microm, with 1-7 tiny polar granules, but without oocyst residuum. The oocyst wall is rough, with 2 layers, approximately 1.4 thick. Ovoidal sporocysts are 11.2 x 7.3 (10-12 x 7-8) microm, with Stieda body and a globular residuum. Sporulated oocysts of a second new species from M. californicus are subspheroidal, 23.1 x 20.7 (20-26 x 19-23) microm, with residuum and 1 polar granule, but a micropyle is absent. The oocyst wall is rough with 2 layers, approximately 1.5 thick. Ovoidal sporocysts are 12.5 x 7.2 (11-14 x 7-8) microm, with a Stieda body and residuum. Sporulated oocysts of a new species from M. ciliolabrum are subspheroidal, 24.9 x 20.1 (18-27 x 17-23) microm, with 1-2 polar granules, but without micropyle and residuum. The oocyst wall is rough with 2 layers, approximately 1.5 thick. Ellipsoidal sporocysts are 12.5 x 9.0 (8-14 x 7 10) microm, with Stieda and substieda bodies and residuum. Sporulated oocysts of a new species from M. evotis are subspheroidal, 21.3 x 18.6 (20-24 x 15-20) microm, with a prominent polar granule, but without micropyle and residuum. The oocyst wall is smooth with 2 layers, approximately 1.0 thick. Ovoidal sporocysts are 12.2 x 8.0 (11-13 x 7.5-9) microm, with Stieda and substieda bodies and residuum. Sporulated oocysts of the new species from N. humeralis are subspheroidal, 22.4 x 18 (21-24 x 17-20) microm, with 1-3 polar granules, but residuum and micropyle are absent. The oocyst wall is lightly sculptured with 2 layers, approximately 1.4 thick. Ovoidal sporocysts are 10.9 x 7.7 (9-12 x 6-8) microm, with Stieda body and residuum. Sporulated oocysts of E. pilarensis Scott and Duszynski, 1997 and those of at least 12 other morphological forms were seen in the other infected bats; these latter forms were seen in too few numbers to be adequately described as new species. PMID- 10386444 TI - Eimeria from bats of Bolivia: two new species from vespertilionid bats. AB - Between 1985 and 1987, fecal samples were collected from 71 bats representing 14 species (Desmodontidae, Molossidae, Noctilionidae, Phyllostomidae, Vespertilionidae) from 8 localities in 3 states (Beni, Pando, Santa Cruz) in Bolivia, South America. Of these, 2 black myotid bats (Vespertilionidae), Myotis nigricans, and 1 tent-making bat (Phyllostomidae), Uroderma magnirostrum, had oocysts in their feces that represent undescribed species of Eimeria. The new species from M. nigricans (2/4, 50%) has sporulated oocysts that are subspheroidal, 18.9 x 16.9 (17-23 x 14-20) microm, without a micropyle; oocyst residuum of 6-8 spheroidal globules and 1 highly refractile polar granule are present. The oocyst wall has 2 layers (approximately 1.3 microm thick), with a rough outer layer. Ovoidal sporocysts are 10.1 x 7.4 (7-14 x 5-10) microm, with a Stieda body, substieda body, and a sporocyst residuum. The new eimerian species from U. magnirostrum (1/2, 50%) has sporulated oocysts that are subspheroidal to ellipsoidal, 23.8 x 20.8 (20-26 x 19-24) microm, without micropyle or oocyst residuum, but 1-3 polar granules are present. The oocyst wall has 2 layers (approximately 1.5 microm thick), with a rough, mammilated outer layer. Ovoidal sporocysts are 11.6 x 8.6 (10-12 x 7-10) microm, with a Stieda body, substieda body and a sporocyst residuum. PMID- 10386445 TI - Larval nematodes (Spiruroidea: Cystidicolidae) in Octopus vulgaris (Mollusca: Cephalopoda: Octopodidae) from the northeastern Atlantic Ocean. AB - Larval nematode parasites (Spiruroidea: Cystidicolidae) are recorded for the first time in Octopus vulgaris Cuvier, 1797 in the northeastern Atlantic Ocean. Prevalence was 16% and mean intensity was 1.46 worms/host. Body length of larval nematodes ranges from 8.3 to 9.3 mm, with a distance from the anterior end to nerve ring from 187.5 to 200 microm, and to excretory pore 194.6-350 microm. Anatomical characteristics, such as deirid, nerve ring, cephalic alae, excretory pore, pseudolabia amphids, sclerotized protuberance, and anus, examined using light microscopy (LM) or scanning electron microscopy (SEM), are illustrated. The nematode was designed as a cystidicolid "Type A" larva. The hemocytic infiltration present in the host tissue around the nematode capsule and the mechanical compression in the infected organs denote the pathogenicity of this nematode. In the study area, O. vulgaris may play the role of an intermediate or paratenic host in the nematode life cycle. PMID- 10386446 TI - Six new nematodes of the Heligmonellidae (Trichostrongylina) collected from endemic murines of Sulawesi, Indonesia. AB - Nematodes in Odilia and Paraheligmonelloides (Trichostrongylina: Heligmonellidae) are first recorded from Sulawesi, Indonesia, with 6 new species from the small intestine of endemic murines: Odilia sulawesiensis n. sp. and Odilia moatensis n. sp. from Rattus xanthurus; Odilia mamasaensis n. sp. and Odilia maxomyos n. sp. from Maxomys musschenbroekii; Paraheligmonelloides eropeplios n. sp. from Eropeplus canus; Paraheligmonelloides paruromyos n. sp. from Paruromys dominator. The 6 species are readily distinguished from congeners in the arrangement and number of the synlophe ridges, dilatation of cuticle, shape of the bursa copulatrix and the spicules, and length ratio of the spicules or the ovejector to body. The intestinal heligmonellid fauna of Sulawesi rats shows affinity to both Sundaland and Australian representatives, reflecting dispersal and speciation history of the nematodes and their hosts. PMID- 10386447 TI - Phylogenetic analysis of Cryptosporidium isolates from captive reptiles using 18S rDNA sequence data and random amplified polymorphic DNA analysis. AB - Sequence alignment of a polymerase chain reaction-amplified 713-base pair region of the Cryptosporidium 18S rDNA gene was carried out on 15 captive reptile isolates from different geographic locations and compared to both Cryptosporidium parvum and Cryptosporidium muris isolates. Random amplified polymorphic DNA (RAPD) analysis was also performed on a smaller number of these samples. The data generated by both techniques were significantly correlated (P < 0.002), providing additional evidence to support the clonal population structure hypothesis for Cryptosporidium. Phylogenetic analysis of both 18S sequence information and RAPD analysis grouped the majority of reptile isolates together into 1 main group attributed to Cryptosporidium serpentis, which was genetically distinct but closely related to C. muris. A second genotype exhibited by 1 reptile isolate (S6) appeared to be intermediate between C. serpentis and C. muris but grouped most closely with C. muris, as it exhibited 99.15% similarity with C. muris and only 97.13% similarity with C. serpentis. The third genotype identified in 2 reptile isolates was a previously characterized 'mouse' genotype that grouped closely with bovine and human C. parvum isolates. PMID- 10386448 TI - Revision of Bursacetabulus (Diplostomidae: Diplostominae) with the proposal of Bursatintinnabulus n. gen., and description of Bursatintinnabus bassanus n. sp. and Bursacetabulus morus n. sp. from northern gannet, Morus bassanus (Aves), from the Texas gulf coast. AB - During a study of digeneans of shorebirds from the Texas gulf coast, 2 undescribed species of diplostomes were found in northern gannet, Morus bassanus. Bursatintinnabulus n. gen. (Diplostomidae) is established with reassignment of Bursacetabulus macrobursus Dronen et al., 1999, as type species, and the second species in the proposed genus is described, Bursatintinnabulus bassanus n. sp. Generic diagnosis of Bursacetabulus Dronen et al., 1999, is emended to include a conical hindbody, an inconspicuous pouchlike or conspicuous well-developed tribocytic organ, and digitiform vitellaria distributed mainly in the hindbody with processes extending into the tribocytic organ and ventrolaterally in the hindbody to the level of the testes. Bursacetabulus morus n. sp. is described as the second species in that genus. In Diplostominae, Bursacetabulus and Bursatintinnabulus n. gen. are most similar to Tylodelphys Diesing 1850, but can be distinguished by having smooth testes and vitellaria that extend ventrolaterally into the hindbody to the level of the posterior testis; and the absence of a genital cone, an acetabulum, and a prepharynx. Bursatintinnabulus n. gen. is different from Bursacetabulus and all other genera of Diplostominae by a well-developed, bell-like skirt surrounding the bursa. PMID- 10386449 TI - Pararhinebothroides hobergi n. gen. n. sp. (Eucestoda: Tetraphyllidea) in Urobatis tumbesensis (Chondrichthyes: Myliobatiformes) from coastal Ecuador. AB - A new species of tetraphyllidean eucestode inhabiting Urobatis tumbesensis from inshore waters of southeastern Ecuador shares 3 synapomorphies with Rhinebothroides spp.: apical bothridial suckers poorly differentiated from the marginal loculi, internal seminal vesicles, and insertion of the vas deferens dorsally closer to the poral than the aporal end of the cirrus sac. The new species differs from Rhinebothroides spp. by lacking medial bothridial septa and loculi and having symmetrical ovarian arms, and possesses an apparent autapomorphic trait by having the vas deferens tapering to a narrow tube before entering the cirrus sac, extending posteriorly to the posterior end of the cirrus sac where it expands into an external seminal vesicle running ventral to the cirrus sac anteriorly to anterior to the vagina. In Rhinebothroides spp., the vas deferens is expanded into an external seminal vesicle near the insertion into the cirrus sac As the sister group of Rhinebothroides, we propose a new genus to accommodate the new species. Phylogenetic evaluation of phyllobothriids recently assigned to Anthocephalum shows that they represent a paraphyletic assemblage of species of varying degrees of relatedness to Rhinebothroides spp. and the new species. Uncovering the relationships of the new species and the various species assigned to Anthocephalum permitted reevaluation of character transformations used in previous phylogenetic analysis of Rhinebothroides. Transformation series for 3 characters, previously based on functional outgroup comparisons, changed and a new character, length of cirrus sac, was added. The new phylogenetic analysis differs from the previous hypothesis only in placing R. scorzai as the sister species of R. circularisi + R. venezuelae + R. moralarai rather than of R. freitasi + R. glandularis + R. mclennanae. The occurrence of the sister species of Rhinebothroides in a Pacific Ocean stingray adds additional support to the hypothesis of Pacific origins of South American freshwater stingrays. PMID- 10386450 TI - Macroevolutionary patterns of male reproductive investment in a clade of parasitic hermaphrodites. AB - The Eucestoda is particularly relevant for questions concerning reproductive investment in male gametes because no other parasitic group displays such diversity in testis size and number within and among species. This diversity has long been used as a valuable taxonomic character, but few researchers have ever investigated its evolutionary significance. In this paper we investigate the evolution of testis number and size within Rhinebothroides (Platyhelminthes: Eucestoda). Our comparative, phylogenetic analysis revealed that overall allocation to male functions, as measured by relative testicular area, does not change within the clade, even though the packaging of that investment in numerous testes is highly variable within, and diverse among, members of the group. PMID- 10386451 TI - Flow cytometric quantification of Toxoplasma gondii cellular infection and replication. AB - The invasion and replication of Toxoplasma gondii are usually analyzed through either optical microscopy or incorporation of tritiated uracil. A new method has been developed using flow cytometric analysis to examine the entry and replication of T. gondii RH strain in Saimiri brain endothelial cells. After cell fixation and permeabilization using saponin, intracellular T. gondii were labeled with a monoclonal antibody against T. gondii SAG-1 (P30; the major cell-surface antigen) followed by fluorescein-conjugated rabbit anti-mouse IgG. The percentage of infected cells obtained using flow cytometry correlated directly with that obtained by UV light microscopy (r = 0.97). The mean fluorescence intensity of infected cells reflects intracellular P30 and assesses intracellular replication. The distribution of fluorescence per infected cell, considered with the percentage of infected cells, also allows a qualitative analysis of replication. Such a method is rapid, easy, and does not require specialized equipment for radioactive labeling. PMID- 10386452 TI - Vertical transmission of Neospora caninum in BALB/c mice determined by polymerase chain reaction detection. AB - Vertical transmission of Neospora caninum was evaluated in BALB/c mice using an N. caninum-specific polymerase chain reaction (PCR) assay as a means of detecting parasite transmission to offspring. BALB/c mice were infected with the NC-1 isolate of N. caninum during pregnancy (days 8-15 gestation). Transmission of parasite, detected by PCR, was determined in 2- to 23-day-old offspring. When dams were infected on days 13-15 of gestation, transfer of parasites was detected in only a proportion of the litter. Infection between days 8 and 12 of gestation resulted in a high frequency of parasite transmission; every offspring from all litters was infected. The tissue locations of parasites in pups of different ages were determined. In young pups (2- to 4-days-old), the predominant sites of infection were the lungs and the brain. In older pups (7- and 23-days-old) the predominant site of infection was the brain. This study shows that PCR may be useful for evaluation of candidate vaccines against horizontal N. caninum infection, vertical transmission, or both. PMID- 10386453 TI - Differentiation of Trichinella genotypes by polymerase chain reaction using sequence-specific primers. AB - A method was developed to identify species and genotypes within the genus Trichinella using polymerase chain reaction (PCR) and specific primers. Enzymatic amplification of 2 partially conserved and repetitive genomic DNA sequences that have been shown to be variable in length within the different Trichinella genotypes form the basis of this test. Within these regions of the genome, 4 sets of primers were evaluated from which 2 were chosen for their ability to differentiate among the genotypes under stringent primer annealing conditions while maintaining high yields of amplification product. Differences in the size of PCR products from multiple isolates of each genotype indicate sufficient variation to identify 7 of the 8 parasite groups within this genus. One primer set can differentiate among some genotypes working from a single larva. Identification of Trichinella genotypes will assist in distinguishing between sylvatic and synanthropic life cycles. Such information will be critical in tracing sources of trichinellosis by easily and unambiguously identifying likely host reservoirs and will provide valuable information for instituting methods of control. PMID- 10386454 TI - Molecular composition of the louse sheath. AB - Flash pyrolysis-gas chromatography/mass spectrometry was used to assess the chemical composition of the head louse's nit sheath. The pyrolyzate of the female insect's secretions, which form a cement-like cylinder holding the egg onto the hair, is dominated by amino acid derivatives and fatty acids. No chitin-specific compounds were detected in the sheath. These results, contrary to previous reports, show that the polymeric complex of the sheath is composed of proteinaceous moieties, possibly cross-linked to aliphatic components. This study constitutes the first chemical characterization of the pyrolysis products of insect (louse) glue and unequivocally confirms that louse sheaths are not chitinous, as suggested by earlier histochemical studies. Development of agents that might loosen nits from the hair shaft is dependent on research that addresses the chemical composition of the nit sheath. PMID- 10386455 TI - Survey for Cyclospora cayetanensis in domestic animals in an endemic area in Haiti. AB - From January 1997 through July 1998, we examined stool samples from 327 domestic animals, including pigs, cattle, horses, goats, dogs, cats, guinea pigs, chicken, ducks, turkeys, and pigeons in Leogane, Haiti, for the presence of Cyclospora cayetanensis infection. No coccidian oocysts morphologically compatible with C. cayetanensis were detected in any of the animal samples, despite their living in, or near, households with infected individuals. These results suggest that domestic animals are not reservoir hosts for C. cayetanensis and that in this endemic area, humans are the only natural host for this parasite. PMID- 10386456 TI - Minicircle variable region probes for characterization of Leishmania (Viannia) species. AB - The minicircle molecules present in the kinetoplast DNA (kDNA) network constitute a particularly useful molecular tool because they are a multicopy target and present a variable region that differs among minicircle classes in the same network. Using the polymerase chain reaction (PCR) and a set of primers directed outwardly from the minicircle conserved region, it is possible to prepare molecular probes representing the pool of variable regions from the different minicircle classes in the kDNA. In order to examine the specificity of the minicircle variable region as hybridization probes in Leishmania (Viannia) species, such fragments were amplified from reference strains and from a panel of isolates representing the zymodeme diversity of Leishmania (Viannia) in Colombia. The size of the amplified products was conserved in Leishmania (Viannia) braziliensis, Leishmania (Viannia) guyanensis, and Leishmania (Viannia) panamensis (650 bp) and diverged in Leishmania (Viannia) equatorensis and Leishmania (Viannia) colombiensis (850 bp). The amplified products were further hybridized to variable region pools of Leishmania braziliensis, Leishmania panamensis, Leishmania guyanensis, and Leishmania equatorensis reference strains. The results obtained from the hybridization experiments support this approach as a means of defining relationships among strains. Hybridization allowed homologies to be perceived, whereas restriction fragment length analysis of the amplified products yielded strain-specific profiles. Apparently, L. (V.) equatorensis and L. (V.) colombiensis minicircle variable regions have no or only low homology with those of other Leishmania (Viannia) species, showing the divergence of those species within the subgenus. PMID- 10386457 TI - Experimental infection of striped marshfrog tadpoles (Limnodynastes peronii) by Ichthyophthirius multifiliis. AB - Ichthyophthirius multifiliis, or white spot, is a well known and widely distributed parasite of freshwater fish. However, it is not know whether it can infect other aquatic vertebrates such as amphibians. This study uses a series of laboratory-based experiments to demonstrate that I. multifiliis can infect the tadpole stage of an amphibian, the striped marshfrog (Limnodynastes peronii) of Eastern Australia. The tadpoles did not appear to develop ichthyophthiriasis at low parasite levels (200 parasites per tadpole), but at high parasite levels (2,000 parasites per tadpole) 100% of the tadpoles developed ichthyophthiriasis. This is the first time that it has been demonstrated that I. multifiliis can infect a nonpiscine vertebrate host. PMID- 10386458 TI - Splenic and hepatic hemozoin in mice after malaria parasite clearance. AB - Hemozoin (malaria pigment) is found in many tissues during malaria infections. In mice that have self-cured from Plasmodium yoelii and Plasmodium chabaudi infections, liver hemozoin concentration and total content decreased for 6-9 mo after parasite clearance. However, both spleen hemozoin concentration and total hemozoin content increased dramatically during this time period. Thus, hemozoin or hemozoin-laden macrophages continue to accumulate in murine spleens for at least several months after malaria parasitemia becomes undetectable. PMID- 10386459 TI - A method for isolation and purification of nematode larvae. AB - A new apparatus for isolating, purifying, and collecting larvae of nematodes is described. Results of collecting larvae with Baermann's traditional method and the pasteur pipette cotton plug method are compared to those obtained with the device in this research note. It is simple, easy to use, and a more effective apparatus for the isolation, purification, and collection of nematode larvae. PMID- 10386460 TI - Rare quadruple malaria infection in Irian Jaya Indonesia. AB - We report an exceptional finding from a blood slide collected in a remote area in the western half of New Guinea Island (Irian Jaya Province, Indonesia). One adolescent patient was found patently coinfected with all 4 known human malaria species, Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, and Plasmodium ovale. Diagnostic erythrocytic stages of all 4 species were clearly seen in the peripheral blood. A nested polymerase chain reaction, using species specific primer pairs to detect DNA, helped substantiate this finding. Previous reports from Africa, Thailand, and New Guinea have detected all 4 species in a population but not simultaneously in an individual with a patent, microscopically detectable infection. We believe this quadruple infection represents the first reported natural case of all 4 human malaria parasites observed concurrently in the peripheral blood from a single Giemsa-stained slide. PMID- 10386461 TI - Hammondia hammondi organelle proteins are recognized by monoclonal antibodies directed against organelles of Toxoplasma gondii. AB - Hammondia hammondi and Toxoplasma gondii, 2 closely related coccidia of cats, are known to share many antigenic molecules as shown by serologic cross reactivity. Monoclonal antibodies (MAbs) directed against the internal organelles of Toxoplasma gondii were tested by immunofluorescence assay and immunoelectron microscopy on the tachyzoites of H. hammondi. The MAbs anti-apex, anti-dense granules, anti-micronemes, and anti-rhoptries recognized, although weakly, the corresponding antigens on H. hammondi. This finding demonstrates that organelles of the 2 parasites are not only morphologically, but also antigenically, similar. PMID- 10386462 TI - Induction of Toxoplasma gondii cystogenesis and multiplication arrest by treatments with a phosphatidylcholine-specific phospholipase C inhibitor. AB - A model of tissue cyst formation was developed using D609, a specific inhibitor of phosphatidylcholine specific-phospholipase C. The phospholipase inhibitor induced a decrease in Toxoplasma gondii multiplication and several successive treatments could lead to an arrest in parasite multiplication and full encystment of the parasites. This could be a first step towards an in vitro model of T. gondii reactivation. PMID- 10386463 TI - Humoral immune response and antigenemia in sheep experimentally infected with Schistosoma bovis. Cross-reactivity with Fasciola hepatica antigens. AB - Circulating antigen level, IgG antibody response to worm antigens and to excretory/secretory products (ES), and specificity to Fasciola hepatica antigens were determined in 6 Schistosoma bovis-infected sheep at weekly intervals for 15 wk. A noninfected control group was included. An enzyme-linked immunosorbent assay (ELISA) sandwich and a double-antibody ELISA test was used for antibody and antigen detection, respectively. The infection induced an early and relatively low IgG response to adult worm extract. This response was significantly elevated by 3 wk postinfection (PI), reached its maximum level at 9 wk PI, and was followed by a subsequent decrease. The response to ES antigens was slightly higher than that to adult worms, although the response started later, at 8 wk PI, and remained at its maximum level until 15 wk. A remarkable level of cross reactivity was observed when adult F. hepatica extract was used. However, a low degree of cross-reactivity was found with ES antigen. The ELISA for circulating antigens was performed at weekly intervals for 8 wk. Antigens were detected as early as the first week of infection, although differences were statistically significant from week 5 onward. The highest values were observed at 7 week PI. PMID- 10386464 TI - Increased parasite abundance associated with reproductive maturity of the clam Anodonta piscinalis. AB - Several studies on vertebrates have demonstrated that reproductive activities may increase the parasite load, but this has not been shown in invertebrate hosts. We studied abundance of a potentially harmful gill parasite, the ergasilid copepod Paraergasilus rylovi, from the freshwater bivalve host Anodonta piscinalis in relation to reproductive maturity of the host in the field. Prevalence of this previously unstudied parasite varied from 90 to 100%, and the mean parasite abundance from 16.3 to 28.8 among 3 study populations. Abundance of P. rylovi increased with host size. In the maturating age groups (3-5 yr) the length adjusted mean parasite abundance among mature, reproducing female clams that brooded glochidia larvae was 2 times higher than in nonreproducing females, the observed pattern being consistent among the 3 study lakes. Alternative, mutually nonexclusive explanations may be found for the result. For example, changes in clam behavior or filtration activity accompanying maturation can increase host exposure to parasites, or reproduction may decrease energy available to host immunologic defense. However, the present result indicates that maturation, and reproduction, is associated with increased parasite abundance in A. piscinalis, an invertebrate host. PMID- 10386465 TI - Robust estimation of parasite component community richness. AB - The performance of 2 nonparametric estimators of species richness, the bootstrap (S(B)) and kth-order jackknife (S(Jk)), are compared using simulated parasite communities. The parameters of the simulation match those of an earlier comparison that favored S(B) as an estimator but did not include S(Jk). S(Jk) is the least biased of the 2 estimators. Whereas the bias of S(B) is significantly affected by true species richness and the proportion of rare species, the bias of S(Jk) is relatively insensitive to changes in these parameters, and is, therefore, recommended as a robust estimator of species richness. PMID- 10386466 TI - Pre- and postnatal testosterone administration induces proliferative epithelial lesions with neuroendocrine differentiation in the dorsal lobe of the rat prostate. AB - BACKGROUND: Androgens are implicated in the pathogenesis of prostatic carcinoma. We have elucidated the role of pre- and postnatal testosterone administration in the occurrence of proliferative lesions as well as neuroendocrine (NE) cells in the rat prostatic complex. METHODS: Female rats were given a single dose of 9 mg testosterone enantate i.m. on day 15 of pregnancy; it gave a high testosterone exposure to the fetus in the early organogenetic period of the rat prostatic complex. One group of the male offspring was followed without further testosterone treatment; a second group received testosterone only in the pubertal period; a third group was given testosterone from puberty and throughout life (46 weeks). These groups were compared to parallel groups (1A-1C) of male offspring without a testosterone supplement in pregnancy. RESULTS: The serum testosterone concentrations in the rats receiving testosterone were significantly higher than those of control rats. Histopathologically, the testosterone-induced proliferative lesions, mainly hyperplastic, were almost exclusively located in the dorsal lobe. Chromogranin A-immunoreactive (CgA-IR) cells were rarely found normally, but occurred more often in the proliferative lesions (P < 0.001). CONCLUSIONS: The incidence of proliferative lesions in rats exposed to testosterone only in puberty was comparable to the incidence found in those rats receiving testosterone in puberty and throughout life. This finding may have clinical implications for young athletes, who use testosterone as an anabolic drug. The occurrence of CgA-IR cells increased in proliferative lesions in the dorsal lobe of the rat prostatic complex. PMID- 10386467 TI - Time for revival of estrogens in the treatment of advanced prostatic carcinoma? Pharmacokinetics, and endocrine and clinical effects, of a parenteral estrogen regimen. AB - BACKGROUND: The present pilot study tested the clinical performance of a new pharmacokinetically guided dosing regimen of parenteral estrogen in patients with advanced prostatic carcinoma. The aim was to accelerate endocrine effects and to avoid cardiovascular side effects. METHODS: Seventeen patients were randomized to intramuscular injections of 240 mg polyestradiol phosphate (PEP) every second week for the first 8 weeks (five doses), followed by a maintenance dose of 240 mg every month; and 16 patients were randomized to bilateral orchidectomy. The estrogen dosing was calculated by pharmacokinetic modelling to achieve a rapid increase in serum estradiol and thereby a fast decrease in testosterone. RESULTS: The predicted increment in serum estrogen was achieved, together with a subsequent decrease in testosterone in the PEP group. In addition, there were no signs of an increased cardiovascular morbidity. This was probably due to a minimal estrogenic influence on the liver and was reflected by unchanged levels of coagulation factor VII. Clinical effects, during the first 2 years of treatment, were similar in the two treatment arms, with 12 patients in the orchidectomy group and 14 patients in the PEP group responding to treatment. CONCLUSIONS: The present parenteral regimen is an efficient and time-saving estrogen regimen with a favorable side-effect profile. PEP seems to offer a potential for revival of the most cost-effective endocrine treatment of cancer of the prostate, i.e., estrogen. PMID- 10386468 TI - Expression of basic fibroblast growth factor and its receptors FGFR1 and FGFR2 in human benign prostatic hyperplasia treated with finasteride. AB - BACKGROUND: The development of benign prostatic hyperplasia (BPH) is an androgen dependent process which may be mediated by a number of locally produced growth factors. One of these, the basic fibroblast growth factor (bFGF or FGF2), has a mitogenic effect on prostatic stroma. High expression levels of bFGF have been reported in BPH. FGFR1 and FGFR2 receptors, that exhibit affinity for bFGF, have been identified in normal and hyperplastic prostate. Finasteride, a 5alpha reductase inhibitor, is an effective drug in the treatment of BPH, inducing regressive changes in the prostate of treated patients, even though its mechanisms of action are not yet completely elucidated. This study was designed to assess the effects of finasteride on the expression levels of bFGF, FGFR1, and FGFR2 in patients with BPH. METHODS: The expression levels of bFGF, FGFR1, and FGFR2 in 9 patients with prostatic hyperplasia treated with finasteride were assessed by immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR) analysis of mRNA expression and were compared with those of 9 control patients with untreated BPH. RESULTS: Immunohistochemistry showed strong bFGF immunoreactivity in the prostatic stroma of untreated patients, this being somewhat weaker in the epithelium. In treated patients, epithelial immunoreactivity was practically negative, and a considerable reduction in stromal immunoreactivity was seen. These findings were also confirmed by RT-PCR. FGFR1 showed a weak immunoreactivity in the stroma and in basal epithelial cells. FGFR1 showed a weak immunoreactivity in the stroma and in basal epithelial cells. FGFR2 exhibited strong stromal immunoreactivity, becoming weaker in the basal epithelium. No differences were seen in the expression of both receptors between the groups of treated and untreated patients. CONCLUSIONS: A marked reduction in bFGF levels is seen in BPH treated with finasteride in comparison to untreated BPH. In our opinion, finasteride may act as a negative regulator of bFGF expression, counteracting the role of bFGF in the development of BPH. PMID- 10386469 TI - Fas antigen/CD-95 upregulation and activation during castration-induced regression of the rat ventral prostate gland. AB - BACKGROUND: Fas antigen/CD 95 is a 45-kDa transmembrane protein that can initiate intracellular signaling pathways, leading to apoptosis when it is clustered on the cell surface. A recent report claiming that the ventral prostate glands of lpr -/- mutant mice (lacking functional fas antigen) do not regress following castration prompted our analysis of the regressing rat ventral prostate gland for evidence that fas antigen might participate in the molecular process leading to prostate cell apoptosis after castration. METHODS: An RNase protection assay and Western blotting analysis were used to quantify fas antigen mRNA and protein expression in the regressing rat ventral prostate gland. Immunoprecipitates of fas antigen from membrane preparations made from control or castrated rat prostates were analyzed for coprecipitation of FADD and RIP proteins to assess the activation state of the fas antigen before and after castration. Finally, prostate tissues obtained from two different strains of lpr -/- mutant mice were analyzed for induced apoptosis after castration by the TUNEL staining method. RESULTS: Rat ventral prostate gland fas antigen mRNA and protein expression was upregulated approximately 3-5-fold in the 3-day castrated rat as compared to hormonally intact rats. Immunoprecipitates of fas antigen from membranes of ventral prostates from castrated rats contained significantly increased amounts of both FADD and RIP proteins when compared to those of intact or control operated rats. However, counts of TUNEL-labeled cells in the ventral prostate glands of castrated lpr -/- mice were not significantly different from those in castrated, genetically normal controls. Likewise, the morphology of apoptotic bodies formed in the prostates of castrated lpr -/- mice was indistinguishable from that in control animals. CONCLUSIONS: Fas antigen/CD-95, a protein that is involved in some forms of apoptosis, is upregulated during regression of the rat ventral prostate gland and becomes functionally "activated." However, our inability to distinguish any difference in the apoptosis rate or in the morphology of the apoptotic bodies formed in response to castration between lpr /- mice and genetically normal controls indicates that, contrary to the prior report, functional fas protein is not required for castration-induced prostate cell apoptosis. PMID- 10386470 TI - Advanced prostate cancer: course, care, and cost implications. AB - BACKGROUND: If prostate cancer screening proves to be effective, some cases will be prevented from reaching the advanced stage. In order to evaluate screening programs thoroughly, it is important to quantify course, care, and accompanying costs of advanced disease. METHODS: We studied 70 files of patients in two hospitals, who had received a diagnosis of distant metastases of prostate cancer and who had died in the years 1994-1998. The total healthcare received by these patients, including symptoms and complaints, was recorded. RESULTS: The most frequently reported symptoms were pain (42%), urogenital symptoms (25%), and malaise (20%). Eighty-nine percent of all patients were hormonally treated (either by orchidectomy and/or chemical castration), and 47% received one or more series of radiation therapy. Sixty-nine percent of all patients were treated with pain medication. The average duration of advanced disease in all patients was 24 months. Average costs of advanced disease were estimated at $11,182 over the total period: $1,547 (14%) was allocated to assessment and outpatient care, and $9,635 (86%) to treatment and costs of hospital stay. Almost half of the total costs were determined by hospital stay. CONCLUSIONS: These data give a better understanding of the course, care, and costs of advanced prostate cancer. These estimates, together with the effects of advanced prostate cancer on quality of life, will be used for the evaluation of prostate cancer screening. PMID- 10386471 TI - Finasteride in association with either flutamide or goserelin as combination hormonal therapy in patients with stage M1 carcinoma of the prostate gland. International Prostate Health Council (IPHC) Trial Study Group. AB - BACKGROUND: It was very reasonable to consider that the combination of the 5alpha reductase, finasteride, and a pure antiandrogen such as flutamide should provide an effective form of maximal androgen blockade (MAB). Finasteride decreases intraprostatic levels of 5alpha-dihydrotestosterone (DHT), and the antiandrogen would restrain the biological action of the residual DHT by interfering with its association with androgen receptor. This form of MAB should sustain the concentration of testosterone in plasma, thereby maintaining sexual function and reasonable quality of life. In order to investigate this, a randomized multicenter phase II clinical trial of patients with untreated M1 cancer of the prostate was developed and undertaken. METHODS: Patients were randomly allocated to one of three treatment schedules: 1) goserelin, 3.6 mg, s.c., monthly in combination with flutamide, 250 mg., t.i.d. and a placebo, daily, in the image of 2 x 5 mg finasteride; 2) goserelin, 3.6 mg., s.c., monthly in combination with finasteride, 10 mg (2 x 5 mg, daily) and a placebo (t.i.d.) in the image of flutamide; and 3) finasteride, 10 mg (2 x 5 mg, daily) in combination with flutamide (250 mg, t.i.d.). The reduction in concentration of serum PSA at 24 weeks was the endpoint of interest. RESULTS: Baseline prostate-specific antigen (PSA) levels of the patients in the three groups were very similar. There was a substantial decrease in levels of PSA in the three groups prior to the end of the study, the percent decrease in the groups being: 1) goserelin and flutamide combination, 99.1% (95% Confidence interval (CI), 97.7, 99.6); 2) goserelin and finasteride combination, 98.75% (95% CI, 97.1, 99.5); and 3) finasteride and flutamide combination, 97.6%, 95% CI, 94.5, 98.9). In the Generalized linear model (GLM) analysis, there was no center by treatment group interaction (P = 20), and there were no significant differences between centers (P = 0.059) nor among the three treatment groups (P = 0.16). CONCLUSIONS: The decrease in levels of PSA in such a group of patients with M1 cancer of the prostate over a 24-week period was surprisingly large, and the differences in these decreased levels between the three treatment arms were remarkably small. There were no apparent differences in bone scan scores, World Health Organization (WHO) performance status, and pain scores between the arms. With regard to sexual function associated with quality of life, there were the understandable difficulties of data collection from patients treated with goserelin. PMID- 10386472 TI - Estrogen receptor-beta: implications for the prostate gland. AB - Estrogens can have profound effects on prostate growth and differentiation. These effects were thought to be mediated by the classical estrogen receptor; however, the discovery of a second estrogen receptor has redefined the estrogen signaling pathway and may have broad implications on estrogen-responsive tissues, including the prostate. The new estrogen receptor, named estrogen receptor-beta (ERbeta), is preferentially expressed in the prostate and maintains some characteristics that are different from ERalpha. Establishing the distribution and function of ERbeta in the various estrogen-responsive tissues is critical to defining its pharmacological and physiological impact. Differential expression of ERbeta may facilitate development of tissue-specific estrogen agonists and antagonists, a goal in the treatment of diseases in estrogen-sensitive tissues such as breast cancer. This article reviews the current knowledge on ERbeta and its potential impact on the prostate. PMID- 10386473 TI - Follow-up evaluation of a phase II prostate cancer vaccine trial. AB - BACKGROUND: A phase II trial, involving infusions of autologous dendritic cells (DC) and two human histocompatibility antigen (HLA-A2)-specific prostate-specific membrane antigen (PSMA) peptides, was recently completed. Thirty percent of the participants, including subjects with hormone-refractory metastastic disease, and those with suspected local recurrence of prostate cancer, were identified as clinical responders. This report describes the follow-up evaluation of 19 responders in the two study groups. METHODS: After conclusion of the study, study participants were subjected to follow-up evaluations at 6-8-week intervals. Each responder was reevaluated for response status, and duration of response was determined. RESULTS: Subjects were observed for an average of 291 days (metastastic group, group A-2) and 557 days (local recurrence group, group B), which included the treatment and follow-up periods. The average duration of response was 149 days for group A-2, and 187 days for group B. A majority of responders (11/19; 58%) were still responsive at the end of the current follow up. CONCLUSIONS: The responses observed may be significant and relatively durable. This study suggests that DC-based cancer vaccines in the future may provide an additional therapy for advanced prostate cancer. PMID- 10386474 TI - Quebec randomized controlled trial on prostate cancer screening shows no evidence for mortality reduction. PMID- 10386475 TI - Reply to Labrie et al. Results of the mortality analysis of the Quebec Randomized/controlled trial (RCT) PMID- 10386476 TI - Determining the risk of bleeding in warfarin therapy. PMID- 10386477 TI - The effect of ACE inhibitors on cardiovascular morbidity and mortality. PMID- 10386478 TI - Salmeterol for nocturnal asthma. PMID- 10386479 TI - An alternative treatment for low back pain. PMID- 10386480 TI - Preventing delirium in hospitalized older patients. PMID- 10386481 TI - A meta-analysis of the treatment of intermittent claudication. PMID- 10386483 TI - Does sibutramine keep the weight off? PMID- 10386482 TI - Bupropion or patch for smoking cessation? PMID- 10386484 TI - On studying effectiveness. PMID- 10386485 TI - Zafirlukast in clinical practice: results of the Accolate Clinical Experience and Pharmacoepidemiology Trial (ACCEPT) in patients with asthma. AB - BACKGROUND: Zafirlukast is an oral leukotriene receptor antagonist used in the treatment of patients with mild to moderate asthma. To investigate its effects in a clinical practice setting, we evaluated zafirlukast in a heterogeneous group of patients who had asthma of different degrees of severity and who were receiving concomitant asthma medications. METHODS: A total of 3759 patients were enrolled at 924 sites. Patients received zafirlukast 20 mg twice a day for 4 weeks. Pulmonary function was measured twice a day, and overall asthma symptom scores, number of nighttime awakenings, severity of morning asthma symptoms, and beta2 agonist use were recorded daily. RESULTS: In the efficacy analysis (3207 evaluable patients), all parameters showed statistically significant improvement that continued throughout the 4 weeks of the trial. A total of 71% of patients had improved pulmonary function and 72% had improved asthma symptoms. Improvement was consistent regardless of asthma severity category and regardless of concomitant asthma medication category. More than 70% of both physicians and patients indicated there was clinical improvement in pulmonary measures as well as in asthma symptoms. Common adverse events reported were headache (3.7%), nausea (1.4%), pharyngitis (1.4%), and sinusitis (1.1%). CONCLUSIONS: Zafirlukast 20 mg twice a day is well tolerated and improves pulmonary function and asthma symptoms, regardless of asthma severity category and regardless of concomitant asthma medication category. PMID- 10386486 TI - Families at the bedside: an ethnographic study of vigilance. AB - BACKGROUND: Vigilance, the close protective involvement of family members with hospitalized relatives, is a relatively recent phenomenon in the hospital setting. Before the 1960s, hospital visiting policies restricted the presence of family members at the bedside. Policies changed during the 1960s and 1970s when health care professionals recognized that parents' staying with their hospitalized children was beneficial for both the parents and the children. Vigilance later became a phenomenon that included family members staying with adult patients. METHODS: Two ethnographic studies were conducted to examine the meanings, patterns, and day-to-day experience of vigilance. Sixteen family members, described by the nursing staff as staying with the patient, participated in informal semistructured interviews. Participant observation was also used in data collection. RESULTS: Data analysis yielded 5 categories of meaning that describe the experience of vigilance: commitment to care, emotional upheaval, dynamic nexus, transition, and resilience. CONCLUSIONS: Managed care, shortened hospital stays, and cost containment make early involvement of the family in the patient's care imperative. An understanding of the family's needs and experiences is prerequisite to that involvement. The categories of meaning discovered in this research can help health care providers understand family members' experience of vigilance. The implications for the family physician include sensitization and awareness of family members' experiences and the developing of specific actions and interactions fostering a commitment to family-centered care that extends to the hospital setting. PMID- 10386487 TI - Intimate partner violence against women: do victims cost health plans more? AB - BACKGROUND: Previous studies of intimate partner violence have not compared the health care costs of female victims with those of a general female population. METHODS: Our study is an analysis of the computerized cost data for 126 identified victims of intimate partner violence in a large health plan in Minneapolis and St. Paul, Minnesota, in 1994. Data were compared with a random sample of 1007 general female enrollees (aged 18 to 64 years) who used health care services in the same year. RESULTS: We found that an annual difference of $1775 more was spent for victims of intimate partner violence than on a random sample of general female enrollees. Regression analyses found that victims of intimate partner violence were significantly younger and had more hospitalizations, general clinic use, mental health services use, and out-of-plan referrals. Use of emergency room services was the same across groups. CONCLUSIONS: Women who were victims of intimate partner violence cost this health plan approximately 92% more than a random sample of general female enrollees. Contrary to the findings of other studies, use of emergency room services was not a driving factor in the higher costs. Findings of significantly higher mental health service use are supported by other studies. PMID- 10386488 TI - Advertisement-induced prescription drug requests: patients' anticipated reactions to a physician who refuses. AB - BACKGROUND: Drug manufacturers increasingly encourage patient prescription drug demand through the use of direct-to-consumer (DTC) advertisements. We describe patients' forecasts of their reactions if their doctor were to deny an advertisement-motivated drug request and then identify significant predictors of these reactions. METHODS: We conducted a random phone survey of 329 Sacramento adults (response rate = 69%). Key outcomes were respondents' perceived likelihood of reacting to the nonfulfillment of a prescription request by becoming disappointed, trying to persuade the physician to reconsider, seeking a prescription from a different physician, and changing physicians. We also assessed associations between the likelihood of these reactions and respondents' evaluations of their physician's communication skills; attitudes toward, assumptions about the regulation of, and past responses to DTC advertising; health status; and demographic characteristics. RESULTS: Disappointment was the most likely reaction (46%). One fourth of the respondents anticipated resorting to persuasion and seeking the prescription elsewhere, while only 15% considered terminating their relationship with their physicians. Subjects who anticipated reacting in these 4 ways reported lower satisfaction with their physicians, evaluated DTC advertising more favorably, and possessed more confidence in the government's regulation of these advertisements. CONCLUSIONS: A sizable fraction of patients believed they would react negatively if their physician refused to provide a prescription for a drug advertised in the general media. Avenues for dealing effectively with patients' advertising-induced requests for prescription drugs are needed. PMID- 10386489 TI - Disclosing complementary and alternative medicine use in the medical encounter: a qualitative study in women with breast cancer. AB - BACKGROUND: Despite recent findings that patients who use complementary and alternative medicine (CAM) typically choose not to mention this to their physicians, little is known about the reasons for this lack of communication. Understanding the reasons for nondisclosure of CAM use is critical to improving physician-patient communication and patient care. METHODS: We are conducting a 5 year prospective cohort study consisting of 4 interview cycles. The multiethnic, population-based sample consists of 86 San Francisco residents with recently diagnosed breast cancer (response rate = 87%). Findings are determined using qualitative analysis of transcribed interviews. RESULTS: At initial contact, 72% of the participants were using at least 1 CAM therapy for breast cancer. Six months later, 65% of participants were using CAM. Of the women being treated by an alternative practitioner, 54% disclosed their CAM use to their physicians. Conversely, 94% discussed details of their biomedical treatments with their alternative practitioner. Reasons for not disclosing CAM use included anticipating the physician's disinterest, negative response, or unwillingness or inability to contribute useful information; the perception that the CAM therapies used were irrelevant to the biomedical treatment course; and the patients' views regarding the appropriate coordination of disparate healing strategies. CONCLUSIONS: Discussions of patients' CAM use are more poorly integrated into the medical encounter than discussions of biomedical treatment are with alternative practitioners. Patients' disclosure is often cautiously modulated, even by those who would welcome an open discussion with their physicians. Specific suggestions for discussing CAM use with patients are presented. PMID- 10386490 TI - What physicians can learn from consumers of dietary supplements. AB - BACKGROUND: Many Americans consume dietary supplements, including vitamins, minerals, herbs, and amino acids. Government regulation of dietary supplements is limited, and patients typically do not consult with their physicians regarding the use of supplements. METHODS: We conducted a qualitative study to describe patients' decisions about the use of dietary supplements and the communication they have with their physicians about such use. Four focus groups of customers from 3 local suburban health food stores were interviewed. RESULTS: The customers in the health food stores we investigated were motivated to pursue wellness and wanted to take responsibility for their health. They would welcome a partnership with their physicians, but generally believed that physicians are closed-minded and have little knowledge about dietary supplements. These consumers determined the effectiveness of dietary supplements through personal study and subjective experimentation. CONCLUSIONS: The health food store customers in our study were self-informed consumers who did not consult their physicians about their use of dietary supplements, because they did not believe that physicians were knowledgeable about or interested in supplements. An open-minded patient-centered approach would help physicians provide better care for patients who use dietary supplements. PMID- 10386491 TI - Interactive computer technology, behavioral science, and family practice. AB - In this paper we discuss conceptual and practical uses for interactive computer applications (ICAs) for family practice, with an emphasis on implications for patient self-management, physician-patient relationships, primary care research, and health care systems quality improvement. We discuss recent behavioral science advances in patient self-management and the advantages and potential limitations of ICAs for medicine. We describe the benefits and risks of using ICAs for providing information, coping-skills training, and social support for patients and for improving the consistency and quality of care given by physicians. There are currently many effective ICAs, and they will play a central role in future health care. There is also the risk of inappropriate use of ICAs. We provide a summary of the empirical literature examining the use of ICAs to aid patients and providers in behavioral change and guidelines adherence efforts. We advise those people researching and applying ICAs in health care to be bold in what they attempt, but cautious in what they claim. Rigorous scientific evaluation and standardized reporting criteria can help quicken this advance, and there are important policy and ethical issues to consider. We conclude with a list of issues for family practices to consider when selecting and using ICAs. PMID- 10386492 TI - Diagnosis of acute bronchitis. PMID- 10386493 TI - Comparing the US and UK health care systems. PMID- 10386494 TI - Advances in the approach to gastroesophageal reflux (GER) and asthma. PMID- 10386495 TI - Gastric asthma: an unrecognized disease with an unsuspected frequency. AB - Bronchial asthma is a disease that has been recognized for centuries, which is influenced mainly by genetic and environmental factors. The current interest of bronchial asthma is focused to ascertain the causes and the mechanisms that induce bronchoconstriction. Recently, abnormalities of the esophageal and gastric tracts have become important related areas for research. In predisposed individuals, these abnormalities can trigger or worsen the particular syndrome better known as "gastric asthma." In bronchial asthma the disorder of gastroesophageal reflux (GER) occurs more often than would be expected by chance. The neurogenic mechanism is considered to be the main cause of bronchoconstriction. The diagnosis of gastric asthma is particularly difficult and it should be considered also when GER is less evident or not recognized. In asthmatic patients the recognition of gastric abnormalities is very relevant for therapeutic problems also when GER is in a subclinical stage. In fact, many drugs used in the treatment of bronchial asthma can promote or enhance GER and subsequently they can worsen the symptoms of gastric asthma. PMID- 10386496 TI - Pulse oximetry in the evaluation of the severity of acute asthma and/or wheezing in children. AB - We evaluated 174 children with acute asthma and/or wheezing attending two different settings, the allergy clinic (AC) and the emergency room (ER), and compared clinical symptoms and spirometric findings with arterial oxygen saturation as measured by pulse oximetry (SpO2). Seventy-four children (4 months to 15 years old) seen at the AC and 100 children (2 months to 14 years old) seen at the ER for the treatment of acute asthma and/or wheezing were evaluated and a clinical score was attributed on the basis of their symptoms. In addition, the heart rate (HR) was recorded and SpO2 was measured. Among the children seen at the AC, 58 were able to perform pulmonary function tests, and the forced respiratory volume in 1 sec (FEV1) and forced expiratory flow between 25% and 75% of the forced vital capacity (FEF(25-75)) were determined. Children from both groups underwent treatment with a nebulized beta2-agonist (Fenoterol 0.5% solution, 0.08 mg/kg/dose, maximum 2.5 mg) and were re-evaluated after 30 min. Our results showed a significant correlation between decrease in clinical scores and increase of SpO2 following treatment with bronchodilator in both groups of children. SpO2 levels correlated positively with FEV1 and FEF(25-75) values, and negatively with clinical scores and heart rate. The data revealed that a clinical score greater than 3 and an SpO2 < 94% were associated with increased severity of the asthma attack. In addition, SpO2 levels < or = 92% were associated with a 6.3 fold greater relative risk for requiring additional treatment. We concluded that determination of oxygen saturation by pulse oximetry is helpful in monitoring the severity of an acute exacerbation of asthma and/or wheezing, and has a prognostic value. PMID- 10386497 TI - Sputum induction as a method of analyzing pulmonary cells: reproducibility and acceptability. AB - Sputum induction has been proposed as a noninvasive method of sampling airway cells for assessing airway inflammation in asthma. Although useful in the research setting, the applicability of this technique to a respiratory clinic, where it might prove useful for clinical management of anti-inflammatory therapy, has not been assessed. We therefore studied the effect of sputum induction in terms of patient acceptability, effect upon airway caliber, and reproducibility of the total cell and differential cell count at 2 weeks in 20 asthmatic subjects first attending an asthma clinic. We compared such patients with normal controls. Thirty-seven subjects underwent sputum induction on two occasions (20 asthmatics and 17 normal subjects) separated by a 2-week interval, using a standardized protocol. Acceptability was assessed by questionnaire. Airway caliber was measured by serial spirometry, using albuterol premedication for asthmatic subjects. Sputum was induced by inhalation of 3.5% saline over 12 min. Total cell and differential counts on induced sputum were assessed and correlated with bronchial hyperresponsiveness, as well as reproducibility of the technique in the clinical setting. All subjects found the process acceptable, although mild side effects occurred in more than 90% of subjects. No differences in acceptability were found between asthmatic and normal subjects. Sputum induction was associated with a significant decrease in forced expiratory volume in 1 sec (FEV1) in normal subjects (from 4.1 +/- 0.17 to 4.02 +/- 0.19 L [p < 0.01] on visit 1 and on visit 2 from 4.01 +/- 0.15 to 3.90 +/- 0.16 L [p < 0.01]), but not in asthmatic subjects after albuterol premedication. Total pulmonary cell yield on the first and second sputum induction days was 1.97 +/- 0.06 x 10(6) cells/mL of sputum and 2.01 +/- 0.05 x 10(6) cells/mL of sputum, respectively, giving a reliability coefficient of 0.77. Less agreement was seen between individual cell differential counts within subjects, but most fell within the expected range on Bland-Altman plots. Sputum induction appears to be safe and acceptable in both normal and asthmatic subjects. A small decrease in FEV1 occurs in normal subjects, which is prevented by albuterol premedication in asthmatics. Reproducibility at 2 weeks yielded similar total numbers of pulmonary cells, but in a clinic population some variability was seen in the number of inflammatory cells, probably because of the small numbers of these cells. This technique may be less valuable in a clinical than a research setting. PMID- 10386498 TI - Socioeconomic factors and asthma hospitalization rates in New York City. AB - Asthma morbidity and mortality are not distributed homogeneously among populations. To assess the relationship between asthma hospitalization rates and socioeconomic factors, we conducted an ecologic analysis using small geographical areas defined by postal zip code in New York City. Asthma hospitalization rates correlated with low median family income, percentage of minorities in the population, and percentage of children under the age of 18. Lack of access to preventive health care, poor housing conditions, environmental exposures, and genetic susceptibility may contribute to high incidence of asthma in some neighborhoods. This report supports the role of socioeconomic factors in asthma and serves to provide data for regulatory and health agencies to concentrate their efforts on neighborhoods most in need. PMID- 10386499 TI - Characteristics and diagnostic significance of spontaneous wheezing in children with asthma: results of continuous in vivo sound recording. AB - The characteristics and diagnostics of wheezing during induced airway obstruction are well documented. The present study addressed (a) the characteristics of spontaneous wheezing with respect to a possible distinction between wheezes during in vivo versus induced airway obstruction, and (b) the relationship between in vivo wheezing and fluctuations in peak expiratory flow (PEF). Tracheal sounds were continuously recorded from 50 children and adolescents with asthma and 10 without asthma in the home environment. Wheezes underwent a qualitative analysis, including their concomitant sound frequencies. Presence of wheezing was scored by two examiners independently and was related to PEF. Spontaneous wheeze varied from solitary rhonchi to prolonged rhythms of loud stridor, and resembled the "induced" wheezes recorded previously. Power spectra showed that the spectral contents (frequency distribution) were comparable, although the in vivo patterns were more prolonged in duration. The diagnostic sensitivity and specificity of wheezing for a reduction in PEF of >20% were 88% and 92%, respectively. It was concluded that in vivo wheeze resembled induced wheeze and was a diagnostically reliable symptom with respect to asthma exacerbations. PMID- 10386500 TI - Computer-based models to identify high-risk adults with asthma: is the glass half empty of half full? AB - This study developed and evaluated the performance of prediction models for asthma-related adverse outcomes based on the computerized hospital, clinic, and pharmacy utilization databases of a large health maintenance organization. Prediction models identified patients at three- to four-fold increased risk of hospitalization and emergency department visits, and were valid for test samples from the same population. A model that identified 19% of patients as high risk had a sensitivity of 49%, a specificity of 84%, and a positive predictive value of 19%. We conclude that prediction models that are based on computerized utilization data can identify adults with asthma at elevated risk, but may have limited sensitivity and specificity in actual populations. PMID- 10386501 TI - Unlimited opportunities for environmental interventions with inner-city asthmatics. AB - The purpose of this study was to identify the asthmatics living in the Lower West Side (LWS) of Buffalo, New York, and then explore the relationship between urban asthmatic and nonasthmatic exposures to many common household aeroallergens. Eight hundred twenty-eight households were visited and 167 asthmatics and 161 nonasthmatics were identified for comparison. Specific self-reported household exposure prevalences were identified for environmental tobacco smoke, sources of molds, household pets, rats, cockroaches, and sources of dust. Sources of molds, pets, and cockroaches were more likely to be found in the homes of asthmatics compared to nonasthmatics (p < 0.05). Other aeroallergens studied, although highly prevalent, were not more likely to be found in either asthmatic or nonasthmatic homes. PMID- 10386502 TI - Adherence to asthma guidelines in general practices. AB - Adherence to asthma practice guidelines is low. Improved compliance could potentially improve care of patients with asthma. The purpose of this study was to determine if patients managed in a general practice with an associated asthma clinic are more likely to use asthma medications according to clinical practice guidelines than patients managed in the general surgery of the practice. A cross sectional study of adult asthmatics, aged 18-55 years, was conducted in six British general practices. Prescription data on all asthma medication was collected for a 6-month period. Information on asthma clinic attendance, age, sex, employment status, other medical illness, and how patients used their inhaled beta2-agonist was collected through questionnaire. The prescription data for asthma medication and patient use of inhaled beta2-agonist were compared to the British Thoracic Society's (BTS) Guidelines for Management of Asthma in Adults to determine if the patient's asthma medication regimen was appropriate. There was no significant association found between appropriate asthma medication and asthma clinic attendance or other patient characteristics. Adherence to the BTS guidelines was low. Fifty-eight percent of the asthma patients used asthma medication regimens that were not consistent with the BTS guidelines published 1 year earlier. Adherence to the BTS guidelines was low regardless of patient characteristics, including asthma clinic attendance, age, sex, employment status, other medical illness, or individual practice. These findings underscore the need to document the utility of clinical practice guidelines which may improve physician compliance. PMID- 10386503 TI - Antioxidant therapy for coronary artery disease: don't paint the walls without treating the termites! PMID- 10386504 TI - Treatment and prevention of sudden cardiac death: effect of recent clinical trials. AB - Tremendous strides have been made in recent years in the treatment and prevention of sudden cardiac death. Large scale trials have now established several interventions that may improve survival in patients susceptible to sudden cardiac death. In patients who have had a sustained ventricular tachyarrhythmia, the current therapy of choice is an implantable cardioverter defibrillator. For prophylaxis of sudden cardiac death in patients without a previous event, several approaches should be considered. Beta-Adrenergic blocking agents are an effective pharmacologic therapy in patients following myocardial infarction, and their efficacy has also most recently been demonstrated in patients with congestive heart failure. There is no Vaughan Williams class I or III antiarrhythmic drug that has demonstrated efficacy as a prophylactic agent to reduce mortality in these populations, with the possible exception of amiodarone. The best therapeutic approach for prophylactic therapy to prevent sudden cardiac death appears to be the implantable cardioverter defibrillator; however, its use can be justified only in patients at high risk for developing sudden cardiac death. Further work is needed to identify the high risk populations in which this therapy is warranted. PMID- 10386506 TI - Pulmonary mucormycosis: the last 30 years. AB - Pulmonary mucormycosis is relatively uncommon but an important opportunistic fungal infection in immunocompromised persons. The literature on the subject is sparse. We describe a recent case and review the literature to delineate the clinical characteristics of this infection. We searched the MEDLINE database for articles published in the English-language literature since 1970 and carefully analyzed 87 cases. The main risk factors were diabetes mellitus, hematologic cancers, renal insufficiency, and organ transplantation. Several patients had no apparent immune compromise. There was a predilection for involvement of the upper lobes. Air crescent signs on chest x-ray films were predictors of pulmonary hemorrhage and death from hemoptysis. Fiberoptic bronchoscopy was a useful diagnostic method, and histopathologic examination was more sensitive than fungal cultures. The overall survival rate was 44%. Patients treated with a combined medical-surgical approach had a better outcome than patients who did not undergo surgery. Thus, this relatively rare but often fatal disease should be suspected in immunocompromised patients who fail to respond to antibacterial therapy. Early recognition and aggressive management are warranted to maximize chances for cure. Optimal therapy requires systemic antifungal therapy, surgical resection, and, when possible, control of the patient's underlying disease. PMID- 10386505 TI - Laboratory diagnosis of vitamin B12 and folate deficiency: a guide for the primary care physician. AB - At one time, the diagnosis of a deficiency of vitamin B12 or folate was considered to be relatively straightforward. As knowledge has accumulated, the limitations of such tests as serum vitamin level measurements and the Schilling test have become apparent. With the development of newer tests, atypical and subclinical deficiency states have been recognized. In this review, available tests used in the diagnosis of vitamin B12 and folate deficiency are discussed, and a rational approach to the diagnosis of these deficiency states is presented. PMID- 10386507 TI - Vitamin E and coronary artery disease. AB - Various studies have evaluated the antioxidant effects of vitamin E in the prevention or treatment of coronary artery disease (CAD). In vitro data suggest that vitamin E protects against oxidation of low-density lipoprotein and decreases the deposition of atherogenic oxidized low-density lipoprotein in arterial walls. Various observational and epidemiological studies also suggest a relationship between vitamin E serum concentrations or intake and CAD. One prospective, randomized trial suggested that low-dosage vitamin E supplementation (50 IU/d) decreases the risk of angina in patients without previously diagnosed CAD. Another study, using high-dosage vitamin E supplementation (400 or 800 IU/d), demonstrated a decrease in the combined end point of nonfatal myocardial infarction and cardiovascular death in patients with established CAD. Discordant data, however, have been published that imply no cardiovascular benefit of low dosage vitamin E supplementation (50 IU/d) and detrimental effects if vitamin E is combined with beta carotene. At this point, clinicians should emphasize a low fat diet with high intake of fruits and vegetable sources containing vitamin E. Supplemental vitamin E may be considered in patients at high risk for CAD or with documented CAD, but the potential beneficial effects should be weighed against possible long-term adverse effects. If vitamin E supplementation is initiated, the literature suggests dosages of 100 to 400 IU/d, with the higher dosage considered in patients with documented CAD. Additional investigation is warranted to further define the role of vitamin E supplementation in CAD and to critically evaluate the optimal dosage, duration of use, and method of consumption (dietary vs supplemental). PMID- 10386508 TI - Bleeding during warfarin and aspirin therapy in patients with atrial fibrillation: the AFASAK 2 study. Atrial Fibrillation Aspirin and Anticoagulation. AB - BACKGROUND: Treatment with warfarin sodium is effective for stroke prevention in atrial fibrillation but many physicians hesitate to prescribe it to elderly patients presumably because of the associated risk for bleeding and the inconvenience of frequent blood tests for the patients. METHODS: In the Second Copenhagen Atrial Fibrillation, Aspirin, and Anticoagulation (AFASAK 2) Study, we studied the rate of bleeding events associated with the incidence of thromboembolic events in patients receiving warfarin sodium, 1.25 mg/d; warfarin sodium, 1.25 mg/d, plus aspirin, 300 mg/d; aspirin, 300 mg/d; or adjusted-dose warfarin therapy aiming at an international normalized ratio of the prothrombin time ratio (INR) of 2.0 to 3.0. The study was scheduled for 6 years from May 1, 1993, but owing to evidence of inefficiency of low-intensity therapy plus aspirin from another study it was prematurely terminated on October 2, 1996. Minor and major bleeding events were recorded prospectively. The rate of bleeding was calculated using the Kaplan-Meier method and risk factors were identified by the Cox proportional hazards model. RESULTS: Of 677 included patients, 130 (median age, 77 years; range, 67-89 years) experienced bleeding. One woman and 12 men experienced major bleeding. Four had intracranial bleeding: 2 cases were fatal and 2 were nonfatal. During treatment with mini-dose warfarin, warfarin plus aspirin, aspirin, and adjusted-dose warfarin, the annual rate of major bleeding was 0.8%, 0.3%, 1.4%, and 1.1%, respectively (P = .20). After 3 years of treatment the cumulative rate of any bleeding was 24.7%, 24.4%, 30.0%, and 41.1% (P = .003), respectively. Increasing INRvalue (P<.001) and prior myocardial infarction (P = .001) were independent risk factors for bleeding, whereas increasing age was not. CONCLUSIONS: Fixed mini-dose warfarin and aspirin alone or in combination were associated with both minor and major bleeding. The small number of major bleeding events in patients receiving adjusted-dose warfarin therapy as compared with those receiving less intensive antithrombotic treatments and the finding of no significant influence of age on the risk for bleeding indicate that even elderly patients with atrial fibrillation tolerate adjusted dose warfarin therapy (INR, 2.0-3.0). PMID- 10386509 TI - Comparison of the effects of lean red meat vs lean white meat on serum lipid levels among free-living persons with hypercholesterolemia: a long-term, randomized clinical trial. AB - BACKGROUND: Patients with hypercholesterolemia are often counseled to limit or eliminate intake of red meats, despite evidence that lean red meats (LRMs) are not hypercholesterolemic in comparison with lean white meats (LWMs). The objective of this study was to evaluate the long-term effects on serum lipids of incorporating LRM (beef, veal, and pork) vs LWM (poultry and fish) into a National Cholesterol Education Program (NCEP) Step I diet in free-living individuals with hypercholesterolemia. METHODS: Subjects included 191 men and women with a serum low-density lipoprotein cholesterol level of 3.37 to 4.92 mmol/L (130-190 mg/dL) and triglyceride level less than 3.96 mmol/L (350 mg/dL). After a 4-week baseline phase, subjects were counseled to follow an NCEP Step I diet including 170 g (6 oz) of lean meat per day, 5 to 7 days per week. Based on random assignment, subjects were instructed to consume at least 80% of their meat in the form of LRM or LWM. Fasting serum lipid levels were assessed 4, 12, 20, 28, and 36 weeks after randomization. RESULTS: After randomization, mean concentrations of total cholesterol (6.09 mmol/L [235.7 mg/dL] vs 6.08 mmol/L [235.2 mg/dL]) and low-density lipoprotein cholesterol (3.99 mmol/L [154.1 mg/dL] vs4.01 mmol/L [154.7 mg/dL]) were nearly identical in the LRM and LWM groups (1% 3% below baseline) during treatment. Mean triglyceride levels remained similar to baseline values and high-density lipoprotein cholesterol concentrations increased by approximately 2% in both groups. CONCLUSIONS: The NCEP Step I diets containing primarily LRM or LWM produced similar reductions in low-density lipoprotein cholesterol and elevations in high-density lipoprotein cholesterol levels, which were maintained throughout 36 weeks of treatment. PMID- 10386510 TI - Traditional risk factors and subclinical disease measures as predictors of first myocardial infarction in older adults: the Cardiovascular Health Study. AB - BACKGROUND: Risk factors for myocardial infarction (MI) have not been well characterized in older adults, and in estimating risk, we sought to assess the individual and joint contributions made by both traditional risk factors and measures of subclinical disease. METHODS: In the Cardiovascular Health Study, we recruited 5888 adults aged 65 years and older from 4 US centers. At baseline in 1989-1990, participants underwent an extensive examination that included traditional risk factors such as blood pressure and fasting glucose level and measures of subclinical disease as assessed by electrocardiography, carotid ultrasonography, echocardiography, pulmonary function, and ankle-arm index. Participants were followed up with semiannual contacts, and all cardiovascular events were classified by the Morbidity and Mortality Committee. The main analytic technique was the Cox proportional hazards model. RESULTS: At baseline, 1967 men and 2979 women had no history of an MI. After follow-up for an average of 4.8 years, there were 302 coronary events, which included 263 patients with MI and 39 with definite fatal coronary disease. The incidence was higher in men (20.7 per 1000 person-years) than women (7.9 per 1000 person-years). In all subjects, the incidence was strongly associated with age, increasing from 7.8 per 1000 person-years in subjects aged 65 to 69 years to 25.6 per 1000 person-years in subjects aged 85 years and older. Glucose level and systolic blood pressure were associated with the incidence of MI, but smoking and lipid measures were not. After adjustment for age and sex, the significant subclinical disease predictors of MI were borderline or abnormal ejection fraction by echocardiography, high levels of intimal-medial thickness of the internal carotid artery, and a low ankle-arm index. Forced vital capacity and electrocardiographic left ventricular mass did not enter the stepwise model. Excluding subjects with clinical cardiovascular diseases such as prior angina or congestive heart failure at baseline had little effect on these results. Risk factors were generally similar in men and women. CONCLUSIONS: After follow-up of 4.8 years, systolic blood pressure, fasting glucose level, and selected subclinical disease measures were important predictors of the incidence of MI in older adults. Uncontrolled high blood pressure may explain about one quarter of the coronary events in this population. PMID- 10386511 TI - Postural hypotension and postural dizziness in patients with non-insulin dependent diabetes. AB - BACKGROUND: Postural hypotension with a decline of 20 mm Hg or more in systolic blood pressure on standing is considered a potentially dangerous hypotensive response. Postural dizziness is often strongly associated with postural hypotension. However, there is conflicting evidence, and previous studies have been confined to the elderly, not specifically to patients with diabetes. Thus, we evaluated the association between postural hypotension and postural dizziness, and determined the factors most likely related to postural hypotension in patients with diabetes. METHODS: The subjects were 204 consecutive noninsulin dependent patients with diabetes and 408 age- and sex-matched control subjects. Postural hypotension was defined as a decline of 20 mm Hg or more in systolic blood pressure 1 minute after standing. Postural dizziness was any feelings of dizziness, lightheadedness, or faintness that occurred while standing during the examination. RESULTS: The prevalence of postural hypotension and postural dizziness in patients with diabetes was higher than in control subjects. Those patients with both diabetes and postural hypotension were older and had higher supine systolic blood pressures and higher plasma glycosylated hemoglobin and fasting glucose levels. They had higher prevalence of postural dizziness, hypertension, and cerebrovascular disease, and lower standing systolic blood pressures than those without postural hypotension. They also were more often being treated with antihypertensive agents. Only 32.8% of patients with diabetes with postural hypotension suffered from postural dizziness. Postural dizziness, hypertension, cerebrovascular disease, and plasma glycosylated hemoglobin levels were independently associated with postural hypotension in patients with diabetes. CONCLUSIONS: Postural dizziness, glycemic control, hypertension, and cerebrovascular disease were important determinants of postural hypotension in patients with diabetes. Postural hypotension was associated with postural dizziness, but it cannot be determined clinically just from the presence of postural dizziness because the sensitivity for diagnosis of postural hypotension is low. PMID- 10386512 TI - Dry eye and dry mouth in the elderly: a population-based assessment. AB - BACKGROUND: Symptoms of dry eye and dry mouth are common in the elderly and are often debilitating. Previous research on small populations has been inconsistent regarding the contribution to sicca symptoms of autoimmune markers, medication use, and other factors. The objective of this study was to determine the population prevalence of symptoms of dry eye and dry mouth and to evaluate possible risk factors. METHODS: This is a population-based study of 2481 individuals, aged 65 to 84 years, residing in Salisbury, Md, and identified by the Health Care Financing Medicare database. The main outcome measures included information on sicca symptoms, medical history, medication use, and joint examination results collected in a standardized manner. Autoimmune status was assessed in 1200 individuals by measuring antinuclear antibody, rheumatoid factor, and autoantibodies to the soluble nuclear antigens Ro/SS-A and La/SS-B by double immunodiffusion. RESULTS: Approximately 27% of the population reported dry eye or dry mouth symptoms to be present often or all the time and 4.4% reported both. The prevalence of dry mouth (but not dry eye) symptoms increased with age, female sex, and white race. No association of sicca symptoms was found with rheumatoid arthritis, smoking, alcohol consumption, reproductive hormonal status, or the presence of autoantibodies. A strong, dose-response relationship was observed between sicca symptoms and the use of certain medication classes. The proportion of the population prevalence of sicca symptoms attributable to the use of drying medications was estimated at 62% for dry eye and dry mouth and 38% for dry eye or dry mouth symptoms. CONCLUSIONS: Sicca symptoms are common in the elderly, and medication side effects appear to be a major underlying factor. Our results do not indicate an association between autoimmune status and sicca symptoms and do not support immunologic testing in persons with sicca symptoms in the absence of other important systemic features. PMID- 10386513 TI - Radioiodine therapy for multinodular toxic goiter. AB - BACKGROUND: Radiolabeled iodine 131 therapy is used for treatment of multinodular toxic goiter, but long-term follow-up studies are lacking. METHODS: A prospective study of 130 consecutive patients (115 women) treated with 131I for multinodular toxic goiter and followed by evaluation of thyroid volume (determined using ultrasound) and thyroid function variables. RESULTS: The patients were observed for a median of 72 months (range, 12-180 months). Sixty-six patients received antithyroid drug pretreatment; 64 did not. Iodine 131 treatment (3.7 MBq/g thyroid tissue corrected to a 100% 24-hour 131I uptake) was given as a single dose in 81 patients, 2 doses in 38, and 3 to 5 doses in 11. One or 2 treatments cured 119 patients (92%), and 68 (52%) became euthyroid within 3 months after 131I treatment. The median 131I dose was 370 MBq (range, 93-1850 MBq). Forty-nine patients needing more than 131I dose had a reduction in median thyroid volume from 56 mL (range, 21-430 mL) to 44 mL (range, 15-108 mL), representing a 24% reduction related to the insufficient 131I dose. In all patients, the initial median thyroid volume of 44 mL (range, 16-430 mL) decreased to 25 mL (range, 8 120 mL) (P<.005), representing a median reduction of 43%, 24 months after the last 131I dose. Hypothyroidism evaluated using life-table analysis developed in 6% of patients who did not receive antithyroid pretreatment and 20% who did (P<.005) after a median of 42 months (range, 3-60 months), the total hypothyroidism frequency being 14% within 5 years of treatment. CONCLUSIONS: Ninety-two percent of patients with multinodular toxic goiter were cured with 1 or 2 treatments. The thyroid volume was reduced by 43%, with few side effects. Iodine 131 should be the choice of treatment in patients with multinodular toxic goiter. PMID- 10386514 TI - Being sensitive to the specifics of predictive values in the diagnosis of tuberculous pleuritis. PMID- 10386515 TI - Proactive measures needed for health care reform. PMID- 10386516 TI - Work-related carpal tunnel syndrome: fix the jobs; don't blame the workers. PMID- 10386517 TI - Culture clash. PMID- 10386518 TI - The underuse of warfarin treatment in the elderly. PMID- 10386519 TI - Ethics and managed care can coexist with a free market. PMID- 10386520 TI - Monitoring adherence to HIV therapy. PMID- 10386521 TI - Consequences of cortical dysplasia during development in rats. AB - PURPOSE: To determine whether focal cortical dysplasia alters excitability in regions distant to the region of the dysplasia. METHODS: We studied the physiological consequences of cortical dysplasia induced by either one or three freeze lesions at birth. Seizure susceptibility was assessed at age 35 days by amygdala kindling. c-fos immunostaining was performed after kainic acid-induced seizures at 10, 20, or 30 days to evaluate the patterns of neuronal activation. RESULTS: Freeze lesions consistently produced uniform regions of dysplasia. No significant differences in seizure susceptibility, as measured by afterdischarge threshold and kindling rate, were seen between controls and rats receiving either one or three freeze lesions. c-fos activation after kainic acid injection was not observed in the region of the dysplasia. However, rats with freeze lesions at age 30 days demonstrated asymmetric c-fos staining with greater staining in CA1 ipsilateral, than contralateral, to the lesion. CONCLUSIONS: Focal cortical dysplasia results in enhancement of c-fos activation in regions outside the borders of the dysplasia. However, as indicated by kindling rate, the functional consequences of these alterations do not appear to be robust. PMID- 10386522 TI - Structure-pharmacokinetic-pharmacodynamic relationships of N-alkyl derivatives of the new antiepileptic drug valproyl glycinamide. AB - PURPOSE: The purpose of this study was to evaluate the structure-pharmacokinetic pharmacodynamic relationships of a series of N-alkyl and N,N-dialkyl derivatives of the new antiepileptic drug (AED), valproyl glycinamide (VGD). METHODS: The following compounds were synthesized: N-methyl VGD (M-VGD), N,N-dimethyl VGD, N ethyl VGD, N,N-diethyl VGD (DE-VGD), and N,N-diisopropyl VGD. These compounds were evaluated for anticonvulsant activity, neurotoxicity, and pharmacokinetics. RESULTS: After i.p. administration to mice in the maximal electroshock seizure test (MES), DE-VGD had an ED50 value comparable to that of VGD (145 and 152 mg/kg, respectively), whereas in the subcutaneous metrazol test (sc Met) model, M VGD had a slightly lower ED50 than VGD (108 and 127 mg/kg, respectively). After oral administration to rats, M-VGD had an MES-ED50 similar to that of VGD (75 and 73 mg/kg, respectively). Of the N-alkyl VGD derivatives studied, M-VGD had the best pharmacokinetic profile: the lowest clearance (5.4 L/h), the longest half life (1.8 h), and the lowest liver-extraction ratio (14%). N,N-dialkylated VGD derivatives underwent two consecutive N-dealkylations, whereas N-alkylated derivatives underwent a single N-dealkylation process, yielding VGD as a major active metabolite. CONCLUSIONS: M-VGD had the most favorable pharmacodynamic and pharmacokinetic profile of the investigated N-alkyl VGD derivatives. VGD was found to be a major active metabolite of M-VGD and to be less neurotoxic than M VGD. Therefore VGD rather than one of the investigated N-alkyl VGD derivatives should be considered for development as a new AED. PMID- 10386523 TI - Surgical outcome in a group of low-IQ patients with focal epilepsy. AB - PURPOSE: Because a low IQ score indicates global brain damage, several authors consider it a contraindication for resective epilepsy surgery. This study reports the postoperative results of a small group of subaverage-intelligence patients with epilepsy who underwent focal resections. METHODS: We report on 16 patients who underwent focal resections (no callosotomy or hemispherectomy). All had IQ's <85 and were >13 years of age at the time of surgery. Low IQ was psychometrically assessed (mean IQ = 70) and confirmed by the patients' educational/occupational status. Clinical characteristics, findings from the preoperative workup, and the surgical treatment are described in detail. Postoperative outcome was evaluated with respect to seizure relief and cognitive/ socioeconomic development. RESULTS: Three months after surgery, 14 (87%) of 16 patients were completely seizure free, and nine (64%) of 14 were seizure free at the 1-year follow-up. Patients' cognitive abilities and socioeconomic status were mostly unchanged and in some cases improved. Seizure outcome was not related to IQ level, and there was no evidence of multiple epileptic foci in the patients with continued seizures. CONCLUSIONS: A low IQ level does not entail the presence of extended epileptogenic regions or multiple epileptic foci. Seizure-relief rates in our group concurred with the rates in patients of average intelligence, and the cognitive/socioeconomic outcome was favorable. We conclude that focal surgery in intellectually impaired patients can be recommended if the preoperative diagnostics confirm a circumscribed seizure onset. PMID- 10386524 TI - Outcome of epilepsy surgery in the first three years of life. AB - PURPOSE: We analyzed our experience over a 6-year period with early-childhood patients who had undergone epilepsy surgery, and investigated the surgical outcomes. METHOD: We reviewed the medical records of 23 children, ages 0-3 years, who underwent epilepsy surgery between 1991 and 1996. RESULTS: Twenty children had partial seizures; two had infantile spasms; and one had generalized tonic clonic seizures at onset. The mean age at onset of seizures was 4.7 months, and the mean age at time of surgery was 15.3 months. A total of 32 operations (21 focal cortical resections and 11 hemispherectomies) was performed. Five of 12 children with seizures secondary to a neuronal migration disorder had reoperations, including three who ultimately underwent complete hemispherectomy. The pathology consisted of hemimegalencephaly in three patients, focal cortical dysplasia (FCD) in eight, tuberous sclerosis in one, Sturge-Weber syndrome (SWS) in five, infarction in two, low-grade glioma (LGG) in three, and post-herpes simplex virus encephalitis (HSE) in one. The follow-up period ranged from 1 to 6.5 years (mean, 3.2 years) from patients' last operation. The seizure outcome according to Engel's criteria was class I in 12 patients, class II in three, class III in six and class IV in two. CONCLUSIONS: Seizure outcomes after surgery were less favorable in infants with FCD than in those with SWS and LGG. Seizure outcome for the patients with hemispherectomies was excellent, compared with those who had focal cortical resections. PMID- 10386525 TI - [11C]Flumazenil PET in patients with epilepsy with dual pathology. AB - PURPOSE: Coexistence of hippocampal sclerosis and a potentially epileptogenic cortical lesion is referred to as dual pathology and can be responsible for poor surgical outcome in patients with medically intractable partial epilepsy. [11C]Flumazenil (FMZ) positron emission tomography (PET) is a sensitive method for visualizing epileptogenic foci. In this study of 12 patients with dual pathology, we addressed the sensitivity of FMZ PET to detect hippocampal abnormalities and compared magnetic resonance imaging (MRI) with visual as well as quantitative FMZ PET findings. METHODS: All patients underwent volumetric MRI, prolonged video-EEG monitoring, and glucose metabolism PET before the FMZ PET. MRI-coregistered partial volume-corrected PET images were used to measure FMZ binding asymmetries by using asymmetry indices (AIs) in the whole hippocampus and in three (anterior, middle, and posterior) hippocampal subregions. Cortical sites of decreased FMZ binding also were evaluated by using AIs for regions with MRI verified cortical lesions as well as for non-lesional areas with visually detected asymmetry. RESULTS: Abnormally decreased FMZ binding could be detected by quantitative analysis in the atrophic hippocampus of all 12 patients, including three patients with discordant or inconclusive EEG findings. Decreased FMZ binding was restricted to only one subregion of the hippocampus in three patients. Areas of decreased cortical FMZ binding were obvious visually in all patients. Decreased FMZ binding was detected visually in nonlesional cortical areas in four patients. The AIs for these nonlesional regions with visual asymmetry were significantly lower than those for regions showing MRI lesions (paired t test, p = 0.0075). CONCLUSIONS: Visual as well as quantitative analyses of FMZ-binding asymmetry are sensitive methods to detect decreased benzodiazepine receptor binding in the hippocampus and neocortex of patients with dual pathology. MRI-defined hippocampal atrophy is always associated with decreased FMZ binding, although the latter may be localized to only one sub-region within the hippocampus. FMZ PET abnormalities can occur in areas with normal appearance on MRI, but FMZ-binding asymmetry of these regions is lower when compared with that of lesional areas. FMZ PET can be especially helpful when MRI and EEG findings of patients with intractable epilepsy are discordant. PMID- 10386526 TI - Function-specific high-probability "nodes" identified in posterior language cortex. AB - PURPOSE: Posterior, "Wenicke's," language areas have a high degree of between subject variability, as shown by electrical-stimulation mapping. We investigated the possibility of an organized structure in the distribution of posterior language areas. METHODS: Extraoperative subdural grid stimulation was performed on 67 left hemisphere-dominant patients before resective epilepsy surgery during counting, naming, and reading. Intersubject-averaged language maps were generated in which stimulation disrupted only one language function and not the others, or combinations of language functions. RESULTS: Language sites, although highly variable between subjects, were not organized randomly and appeared to be arranged into several focal, non-contiguous, higher probability "nodes" devoted to different aspects of language processing. Speech-arrest sites were concentrated in classic Wernicke's area. Areas where stimulation induced only reading errors were found in the posterior middle temporal gyrus and the inferior parietal lobule. These regions may correspond with an orthographic input lexicon. Areas eliciting only naming errors were found in the posterior inferior temporal gyrus extending into the mid-middle temporal gyrus and may represent a visual representation input lexicon. Sites where stimulation elicited errors in both naming and reading were more variable in location than sites devoted to only one function, extended farther anteriorly along the temporal neocortex, and may correspond with a semantic lexicon. CONCLUSIONS: The existence of high probability nodes in posterior language cortex supports a modality-specific modular architecture and the possibility of a conserved, universal structure. PMID- 10386527 TI - Early and late posttraumatic seizures in traumatic brain injury rehabilitation patients: brain injury factors causing late seizures and influence of seizures on long-term outcome. AB - PURPOSE: To demonstrate risk factors involved in the origin of late posttraumatic seizures (LPTSs) in civilian traumatic brain injury (TBI) rehabilitation patients and the occurrence of LPTSs in this population, as well as the time of the first late seizures, and influence of these seizures on functional and occupational long-term outcome. METHODS: A consecutive sample of 490 patients (age range, 0.8 71 years) with TBI, and with postinjury problems in their education and employment, were followed up for > or =5 years from the time of injury in a rehabilitation and reemployment program. The study was carried out at the outpatient neurologic clinic of the Kauniala Hospital, which specializes in brain injuries in Finland and works in close cooperation with the Department of Clinical Neurosciences at the Helsinki University Central Hospital. Main outcome measures were functional outcome, as measured on the Glasgow Outcome Scale (GOS), and the capacity for employment at the end of follow-up. Outcomes were studied separately among patients with late seizures and for the nonseizure group. RESULTS: Children age 7 years or younger at time of injury more often had early posttraumatic seizures (EPTSs), than did adolescents or adults. The time elapsed between brain injury and the first late seizure also was longer in older age groups. EPTSs and depressed skull fracture had a statistically significant relation to the origin of LPTSs. Permanent posttraumatic neurologic deficit, linear skull fracture, and permanent local brain lesion documented on a computed tomography (CT) scan appeared clinically important as risk factors. Late seizures did worsen the functional outcome but had no significant influence on reemployment at the end of follow-up. CONCLUSIONS: Young children are more prone to early seizures, and adolescents and adults, to late seizures. The main risk factors for LPTSs are early seizures and depressed skull fracture. Severity of brain injury, as measured by a low GCS score, prolonged unconsciousness, and posttraumatic amnesia (PTA) without local brain lesion, should not be considered risk factor for LPTSs. Thorough follow-up of patients with TBI with seizures and adequate antiepileptic therapy may help attain rehabilitation goals and reemployment. PMID- 10386528 TI - New antiepileptic drugs: comparison of key clinical trials. AB - PURPOSE: Data accrued from clinical trials of five new antiepileptic drugs (AEDs) are compared for efficacy in reducing seizures and self-reported adverse events as a basis of selection among new AEDs. Drawbacks to use of these data also are demonstrated. METHODS: A review of double-blind, placebo-controlled clinical trials of a new AED or placebo added to a standard AED provided data on reduction of complex partial seizures (CPSs). Success is > or =50% fewer CPSs with a new AED or placebo; Overall Improvement is the success rate with drug minus the success rate with placebo. Adverse events were tabulated from product-labeling lists of COSTART items (incidence, > or =5%). The Summary Complaint score is the total number of reports of individual events for each AED. RESULTS: Efficacy data demonstrate differences in Overall Improvement rates among five new AEDs and placebos (p = 0.001). However, rates of response to placebo also differed significantly among trials (p = 0.01). Adverse events predominantly affect central nervous system, psychiatric, and general body systems. However, patients in the placebo control groups did not consistently report adverse effects. Summary Complaint scores differ among the five new AEDs, but variability in use of COSTART terms nullifies comparisons. CONCLUSIONS: Comparisons of data for five new AEDs provide information for selection among treatments when a second drug is needed to improve control of CPSs. However, significant differences among the control groups and other problems make comparisons between trials problematic. The final choice should be based on the need of the individual patient for superior seizure control versus minimal adverse effects. PMID- 10386530 TI - Confirmation of two magnetoencephalographic epileptic foci by invasive monitoring from subdural electrodes in an adolescent with right frontocentral epilepsy. AB - PURPOSE: To report our evaluation of interictal two epileptic spike fields on magnetoencephalography (MEG) by using invasive intracranial monitoring in a patient without lesion on magnetic resonance imaging (MRI). METHODS: A 15-year old left-handed boy with a 9-year history of refractory simple partial seizures, secondarily generalized, and a normal MRI, was studied with MEG to define magnetic spike sources, followed by invasive intracranial monitoring with subdural electrodes to delineate the epileptogenic zone and eloquent function pursuant to focal cortical excision. RESULTS: MEG demonstrated two spike foci on the right middle frontal and inferior rolandic areas adjacent to the sensory area. Ictal recordings during prolonged invasive monitoring from subdural electrodes revealed two epileptogenic zones in the same locations as those defined by MEG. Focal cortical excision was performed of each epileptogenic zone. The patient has been seizure free for 24 months without neurologic deficit. CONCLUSIONS: Magnetic source imaging is a valuable adjunct in the planning of subdural grid placement in epilepsy surgery, particularly in patients in whom conventional imaging fails to reveal a lesion. PMID- 10386529 TI - Lamotrigine monotherapy in newly diagnosed untreated epilepsy: a double-blind comparison with phenytoin. AB - PURPOSE: Lamotrigine is an effective add-on therapy against a range of epileptic seizure types. Comparative studies with carbamazepine (CBZ) as monotherapy in newly diagnosed epilepsy suggest similar efficacy. In this study, lamotrigine (LTG) and phenytoin (PHT) are compared. METHODS: In a double-blind parallel groups study, 181 patients with newly diagnosed untreated partial seizures or secondarily or primary generalised tonic-clonic seizures were randomised to two treatment groups. One group (n = 86) received LTG titrated over 6 weeks from a starting dose of 100 mg/day. The other (n = 95) received PHT titrated from 200 mg/day. Treatment continued for < or =48 weeks. RESULTS: The percentages of patients remaining on each treatment and seizure free during the last 24 and 40 weeks of the study, and times to first seizure after the first 6 weeks of treatment (dose-titration period), did not differ significantly between the treatment groups. These were measures of efficacy. Time to discontinuation, a composite index of efficacy and safety, likewise did not distinguish between treatments. Adverse events led to discontinuation of 13 (15%) patients from LTG and 18 (19%) from PHT. The adverse-event profile for LTG was dominated by skin rash [discontinuation of 10 (11.6%) patients compared with five (5.3%) from PHT] rather than central nervous system side effects: asthenia, somnolence, and ataxia were each significantly more frequent in the PHT group. The high rate of rash with LTG was probably due to the high starting dose and may be avoidable. A quality-of-life instrument, the SEALS inventory, favoured LTG. Patients taking PHT showed the biochemical changes expected of an enzyme-inducing drug, whereas those taking LTG did not. CONCLUSIONS: LTG and PHT monotherapy were similarly effective against these seizure types in patients with newly diagnosed epilepsy. LTG was better tolerated, more frequently causing rash, but with a lower incidence of central nervous system side effects. PMID- 10386531 TI - Idiopathic rolandic epilepsy with "interictal" facial myoclonia and oromotor deficit: a longitudinal EEG and PET study. AB - PURPOSE: The prognosis of benign epilepsy with centrotemporal spikes (BECTS) is always favorable as far as the epilepsy is concerned. However, some data suggest that affected children may be at risk for minor cognitive impairment. We report here the longitudinal study of a young girl demonstrating that BECTS also may be associated with severe motor disturbances. METHODS: BECTS (rare left oromotor seizures, right rolandic spike-waves activated during sleep) started at the age of 3 years 6 months in a girl with normal initial psychomotor development. Her clinical, neuropsychological, and EEG status was assessed every 3-6 months. Regional cerebral glucose metabolism was measured by using the [18F]fluorodeoxyglucose-positron emission tomography (FDG-PET) method. RESULTS: Between the age of 5 and 6 years, the girl had (a) increased seizure frequency; (b) brief perioral and palpebral myoclonic jerks, concomitant with the spike component of interictal spike-waves, and (c) persistent but fluctuating oromotor deficits (drooling, dysarthria, dysphagia). The EEG showed a marked increase in abundance and amplitude of wake and sleep interictal abnormalities, which became bilateral. Awake FDG-PET revealed a bilateral increase of glucose metabolism in opercular regions. A complete and definitive EEG and clinical remission occurred at age 5 years 11 months and has persisted since (present age, 7 years 9 months). CONCLUSIONS: This case confirms that during BECTS, epileptiform dysfunctions within rolandic areas may induce "interictal" positive or negative oromotor symptoms, independent of classic seizures. PMID- 10386532 TI - Early-onset benign childhood occipital seizure susceptibility syndrome: a syndrome to recognize. AB - Early-onset benign childhood occipital seizures (EBOS) described by Panayiotopoulos constitute the commoner after the rolandic phenotype of a childhood seizure susceptibility syndrome. EBOS are the clinical representative of occipital spikes. Their cardinal features are infrequent (often single) partial seizures manifested with deviation of the eyes and vomiting, frequently evolving to hemi- or generalized convulsions. Ictal behavioral changes, irritability, pallor, and rarely cyanosis, and eyes wide open are frequent. Retching, coughing, aphemia, oropharyngolaryngeal movements, and incontinence may occur. Consciousness is usually impaired or lost, either from the onset or the course of the fits, but in a few children, it may be preserved. Duration varies from a few minutes to hours (partial status epilepticus). Seizures are usually nocturnal, but semiology is similar in nocturnal or diurnal fits. Onset is between 1 and 12 years with a peak at 5 years. One third of children have a single seizure, the median total number of fits is two to three, and the prognosis is invariably excellent, with remission usually occurring within 1 year from onset. A few children may later develop rolandic or other benign partial seizures. The likelihood to have seizures after age 12 years is exceptional and rarer than that of febrile convulsions. EEG shows occipital paroxysms demonstrating fixation-off sensitivity, but random occipital spikes, occipital spikes in sleep EEG alone, or normal EEG may occur. Centrotemporal and other spike foci may appear in the same or more frequently in subsequent EEGs. The EEG does not reflect clinical course and severity. PMID- 10386533 TI - Prevalence and pattern of epilepsy in India. AB - PURPOSE: To estimate the prevalence of epilepsy in India by meta-analysis of previously published and unpublished studies and to determine patterns of epilepsy by using community-based studies. METHODS: We attempted to identify as many previously published and unpublished studies as possible on the prevalence of epilepsy in India. The studies were assessed with regard to methods and definitions. The prevalence rates for rural and urban populations and for men and women were calculated with a 95% confidence interval (CI). The studies that provided details on age structure, age-specific rates, and patterns of epilepsy were chosen for meta-analysis. Both crude values and age-standardized prevalence rates were calculated after accounting for heterogeneity. RESULTS: Twenty studies were found involving a sample population of 598,910, among whom 3,207 had epilepsy. This resulted in a crude prevalence of 5.35/1,000. After a correction for heterogeneity due to interstudy variation, the overall prevalence per 1,000 (and its 95% CI) was 5.33 (4.25-6.41); with urban areas at 5.11 (3.49-6.73); rural areas, 5.47 (4.04-6.9); men, 5.88 (3.89-7.87); and women 5.51 (3.49-7.53). After correction for the variability in estimates of heterogeneity, age standardized rates (from five studies) revealed that the prevalence rates per 1,000 (and the 95% CI), were as follows: overall, 5.59 (4.15-7.03); men, 6.05 (3.79-8.31); women, 5.18 (3.04-7.32); urban, 6.34 (3.43-9.25); rural, 4.94 (3.12 6.76). Urban men and women had a higher prevalence of epilepsy compared with rural ones, however the difference was not statistically significant. Age specific prevalence rates were higher in the younger age group, with the onset of epilepsy reported mostly in the first three decades of the sample population's lives. The treatment gap (i.e., the percentage of those with epilepsy who were receiving no or inadequate treatment) was more than 70% in the rural areas. CONCLUSIONS: Based on the total projected population of India in the year 2001, the estimated number of people with epilepsy would be 5.5 million. Based on a single study on the incidence of epilepsy, the number of new cases of epilepsy each year would be close to half a million. Because rural population constitutes 74% of the Indian population, the number of people with epilepsy in rural areas will be approximately 4.1 million, three fourths of whom will not be getting any specific treatment as per the present standard. PMID- 10386534 TI - Prevalence of epilepsy in Silivri, a rural area of Turkey. AB - PURPOSE: To learn the prevalence of epilepsy in Silivri, a western town of Turkey, a randomized door-to-door survey was conducted using a standard questionnaire. The method of the study was adopted from the suggestions of the World Health Organization (WHO) for prevalence studies in developing countries, and the criteria were derived from Guidelines for Epidemiologic Studies on Epilepsy proposed by the Commission on Epidemiology and Prognosis, the International League Against Epilepsy (ILAE) 1993. METHODS: From June 1 to October 1, 1994, 4,803 people out of a total population of 70,394 were surveyed. The questionnaire, which was administered by practitioners and intern doctors, consisted of 15 questions, with a sensitivity of 99.9% and a specificity of 76%. After the survey, neurologists examined all of the 415 people suspected of having epilepsy and classified the seizures of the active cases. RESULTS: Of the 415 suspected cases, 49 people (24 women, 25 men) were determined as having epilepsy on the assessment day of October 1, 1994. The crude point prevalence of active epilepsy was 10.2 of 1,000 for the region. The prevalence of active epilepsy among women was 10.01 of 1,000 and among men was 10.39 of 1,000. Of the 49 cases, 40.8% had generalized seizures, 53.1% had partial onset seizures, and 6.1% could not be classified. Only 7.7% of the cases with partial onset seizures were defined as probable symptomatic cases. CONCLUSIONS: Onset of the disease peaked at the first decade of life. On the assessment day, 44.9% of those with epilepsy were receiving treatment, and 65.1% had visited religious figures at the onset or during the course of the disease, a figure that reveals the high prevalence of mystical beliefs about the disease in the study area. PMID- 10386535 TI - Physical exercise in outpatients with epilepsy. AB - PURPOSE: To compare the exercise habits in a sample of adult outpatients with epilepsy with those of a general population of the same age and sex and furthermore to study physical exercise as a seizure precipitant and the risk of sustaining seizure-related injuries while exercising. METHODS: Two hundred four adult outpatients with active epilepsy responded to two questionnaires. The first one, addressing exercise habits, was a selected part of a broad self-assessing screening used every second year by a marketing and media research institute to reveal changes in the average Norwegian's lifestyle. The exercise habits of the epilepsy population were compared with those of the average population. The other questionnaire, addressing seizures and injuries related to physical exercise, consisted of eight sections and was developed at the National Center for Epilepsy in Norway. RESULTS: The portion of those never exercising was significantly higher among the patient group compared with the average population. Otherwise, the exercise patterns were very similar in the two populations. However, the patients exercised more often in fitness centers and together with friends, whereas individual activities like skiing and swimming were more often preferred by the average Norwegian. Of the 204 patients, 53 and 63% had never experienced seizures during or immediately after exercise, respectively. About 10% of the patients claimed that they had seizures quite often in connection with exercise. However, only 2% had genuine exercise-induced seizures, here arbitrarily defined as having seizures in >50% of the training sessions. Among those prone to have exercise-related seizures, there was a predominance of patients with symptomatic localization-related epilepsy (i.e., with an underlying structural brain lesion). Most exercise-related seizures occurred during strenuous activity. About 38% of the patients claimed to have personal experience regarding whether regular physical exercise influenced their seizure disorder; of these, 53% claimed there was no influence, 36% claimed there was a positive influence, and 10% reported a negative influence. Thirty-six percent of the patients had experienced injuries in connection with physical exercise, but in only 10% were these injuries associated with seizures. The injuries were mostly mild. CONCLUSIONS: The surveyed sample of epilepsy outpatients was more active than expected, and their exercise pattern closely resembled that of the average Norwegian population. In the majority of the patients, physical exercise had no adverse effects, and a considerable proportion (36%) claimed that regular exercise contributed to better seizure control. However, in approximately 10% of the patients, exercise appeared to be a seizure precipitant, and this applied particularly to those with symptomatic partial epilepsy. The risk of sustaining serious seizure-related injuries exercising seemed modest. PMID- 10386536 TI - Partial seizure with aphasic speech arrest caused by watching a popular animated TV program. AB - On the evening of December 16, 1997, about 700 children across Japan were hospitalized because of convulsive seizures or vomiting experienced while watching a popular animated TV program that included blue and red stimuli that alternated at 12 flashes per second. In one case, an 11-year-old girl developed a hallucination in the right visual field and a subsequent cramp on the right side of her face, with aphasic speech arrest. She had no history of seizures. Her electroencephalogram (EEG) showed normal background activity and no epileptiform discharges. Intermittent photic stimulation provoked a photoparoxysmal response. Her main clinical manifestation was a TV-induced left occipital lobe seizure spreading toward the left inferior frontal lobe. This suggested a functional link from the occipital lobe to the frontal operculum. PMID- 10386537 TI - Opercular reflex seizures: a case report with stereo-electroencephalographic demonstration. AB - Reflex epileptic seizures of opercular origin have been described previously based on video-electroencephalographic monitoring, but very few patients have been explored with depth electrodes. We report a woman with late-onset epilepsy who had intractable seizures despite trials of several antiepileptic drugs. At the time of the depth-electrode recordings, seizures were usually continuous and occurred either spontaneously or were induced by movements of the jaw and mouth. The seizures originated in the deep central opercular cortex; localization was confirmed by a good surgical outcome. PMID- 10386539 TI - Lithium: proving its mettle for 50 years. PMID- 10386538 TI - Stress-induced immunomodulation: implications for infectious diseases? PMID- 10386540 TI - Probing informed consent in schizophrenia research. PMID- 10386542 TI - Governance plan paves way for search for JAMA editor. PMID- 10386541 TI - Prostate detection possibility. PMID- 10386543 TI - From the Centers for Disease Control and Prevention. Prenatal discussion of HIV testing and maternal HIV testing--14 states, 1996-1997. PMID- 10386544 TI - From the Centers for Disease Control and Prevention. Determination of nicotine, pH, and moisture content of six US commercial moist snuff products--Florida, January-February 1999. PMID- 10386545 TI - Anonymous HIV testing and medical care. PMID- 10386546 TI - Clinical crossroads: a 45-year-old woman with premenstrual dysphoric disorder. PMID- 10386547 TI - Patient self-management of oral anticoagulation. PMID- 10386549 TI - Damages and the expert witness. PMID- 10386548 TI - Abdominal adiposity and risk of heart disease. PMID- 10386550 TI - Poetry and medicine. PMID- 10386551 TI - Meta-analysis of tacrine for Alzheimer disease: the influence of industry sponsors. PMID- 10386552 TI - Relationship of ascorbic acid to blood lead levels. AB - CONTEXT: Some animal studies suggest that orally administered ascorbic acid may chelate lead and decrease the risk of the toxic effects of lead. However, results from several small studies in humans have yielded inconclusive evidence of a beneficial effect of ascorbic acid on lead toxicity. OBJECTIVE: To examine the relationship between serum ascorbic acid levels and prevalence of elevated blood lead levels. DESIGN, SETTING, AND PARTICIPANTS: Cross-sectional analysis of a probability sample of the US population enrolled in the Third National Health and Nutrition Examination Survey, 1988-1994 (4213 youths aged 6-16 years and 15365 adults aged > or =17 years) without a history of lead poisoning. MAIN OUTCOME MEASURES: Elevated and log blood lead levels by serum ascorbic acid level. RESULTS: A total of 22 youths (0.5%) and 57 adults (0.4%) had elevated blood lead levels (defined as > or =0.72 micromol/L [15 microg/dL]) and > or =0.97 micromol/L [20 microg/dL], respectively). After controlling for the effects of age, race, sex, income level, and dietary energy, fat, calcium, iron, and zinc intake, youths in the highest serum ascorbic acid tertile had an 89% decreased prevalence of elevated blood lead levels compared with youths in the lowest serum ascorbic acid tertile (odds ratio, 0.11; 95% confidence interval, 0.04-0.35; P for trend = .002). Adults in the highest 2 serum ascorbic acid tertiles had a 65% to 68% decreased prevalence of elevated blood lead levels compared with adults in the lowest serum ascorbic acid tertile (P for trend = .03). As a continuous predictor, serum ascorbic acid level was independently associated with decreased log blood lead levels among adults (P<.001), but not among youths (P=.14). CONCLUSIONS: Our data suggest that high serum levels of ascorbic acid are independently associated with a decreased prevalence of elevated blood lead levels. If these associations are related causally, ascorbic acid intake may have public health implications for control of lead toxicity. PMID- 10386553 TI - Association of dental caries and blood lead levels. AB - CONTEXT: Experiments show that dental caries rates are higher among lead-exposed animals, but this association has not been established in humans. OBJECTIVE: To examine the relationship between blood lead levels and dental caries. DESIGN: Cross-sectional survey conducted from 1988 to 1994 that included a dental examination and venipuncture blood lead assay. SETTING AND PARTICIPANTS: A total of 24901 persons aged 2 years and older who participated in the Third National Health and Nutrition Examination Survey, which assessed the health and nutritional status of children and adults in the United States. MAIN OUTCOME MEASURES: For children aged 2 to 11 years, the sum of decayed and filled deciduous or primary surfaces; for persons aged 6 years and older, the sum of decayed and filled permanent surfaces; for those 12 years and older, the sum of decayed, missing, and filled surfaces. RESULTS: The log of blood lead level was significantly associated with the number of affected surfaces for both deciduous and permanent teeth in all age groups, even after adjusting for sociodemographic characteristics, diet, and dental care. Among children aged 5 to 17 years, a 0.24 micromol/L (5-microg/dL) change in blood lead level was associated with an elevated risk of dental caries (odds ratio, 1.8; 95% confidence interval, 1.3 2.5). Differences in blood lead level explained some of the differences in caries prevalence in different income levels and regions of the United States. We estimated the population attributable risk of lead exposure to be 13.5% and 9.6% of dental caries occurring in 5- to 17-year-olds exposed to the high and moderate levels, respectively. CONCLUSIONS: Environmental lead exposure is associated with an increased prevalence of dental caries in the US population. Findings may help explain the distribution of caries by income and region of the United States. PMID- 10386554 TI - Chronic hyponatremic encephalopathy in postmenopausal women: association of therapies with morbidity and mortality. AB - CONTEXT: Chronic hyponatremia in postmenopausal women is a common clinical problem often viewed as benign. Fluid restriction is usually the recommended therapy, largely because the extent of morbidity is unknown and because it has been postulated that intravenous (IV) sodium chloride may cause brain damage. OBJECTIVE: To compare IV sodium chloride with fluid restriction in the treatment of postmenopausal women with chronic symptomatic hyponatremia. DESIGN: Nonrandomized prospective study. SETTING: Two university medical centers and affiliated community hospitals. PATIENTS: A total of 53 postmenopausal women with chronic symptomatic hyponatremia (chronic plasma sodium <130 mmol/L in the presence of central nervous system manifestations) treated consecutively from 1988-1997 and followed up for 1 year. The mean (SD) age of the patients was 62 (11) years. INTERVENTIONS: The therapeutic interventions were IV sodium chloride before respiratory insufficiency (n = 17), IV sodium chloride after respiratory insufficiency (n = 22), and fluid restriction only (n = 14). MAIN OUTCOME MEASURES: Morbidity and neurological outcome at 4 months or longer as assessed by cerebral performance category (CPC) in relation to the therapy, initial plasma sodium level, and rate of correction. RESULTS: Chronic symptomatic hyponatremia (mean [SD] sodium level 111 [12] mmol/L) was present for 5.2 [4.5] days. Death or major morbidity occurred in 44 (83%) of 53 patients, including 10 with orthopedic injury. Twelve patients had hypoxemia (PO2 = 63 [25] mm Hg) and cerebral edema. Among patients who received IV sodium chloride before respiratory insufficiency, plasma sodium levels were increased by 22 (10) mmol/L in 35 hours and patients had a CPC of 1.0 (normal or slight disability). Among patients who received IV sodium chloride after respiratory insufficiency, plasma sodium levels were increased by 30 (6) mmol/L in 41 hours and patients had a CPC of 3.0 (1.2) (severe disability). Among patients who had fluid restriction only, plasma sodium levels were increased by 3 (2) mmol/L in 41 hours and patients had a CPC of 4.6 (0.7) (4 = persistent vegetative state; 5 = death). The outcomes did not correlate with either the initial plasma sodium level (r=0.05, P>.12) or the rate of correction (r=0.31, P>.10). CONCLUSIONS: Chronic symptomatic hyponatremia in postmenopausal women can be associated with major morbidity and mortality. Therapy with IV sodium chloride was associated with significantly better outcomes than fluid restriction. PMID- 10386555 TI - Variations in the care of HIV-infected adults in the United States: results from the HIV Cost and Services Utilization Study. AB - CONTEXT: Studies of selected populations suggest that not all persons infected with human immunodeficiency virus (HIV) receive adequate care. OBJECTIVE: To examine variations in the care received by a national sample representative of the adult US population infected with HIV. DESIGN: Cohort study that consisted of 3 interviews from January 1996 to January 1998 conducted by the HIV Cost and Services Utilization Consortium. PATIENTS AND SETTING: Multistage probability sample of 2864 respondents (68% of those targeted for sampling), who represent the 231400 persons at least 18 years old, with known HIV infection receiving medical care in the 48 contiguous United States in early 1996 in facilities other than emergency departments, the military, or prisons. The first follow-up consisted of 2466 respondents and the second had 2267 (65% of all surviving sampled subjects). MAIN OUTCOME MEASURES: Service utilization (<2 ambulatory visits, at least 1 emergency department visit that did not lead to hospitalization, at least 1 hospitalization) and medication utilization (receipt of antiretroviral therapy and prophylaxis against Pneumocystis carinii pneumonia). RESULTS: Inadequate HIV care was commonly reported at the time of interviews conducted from early 1996 to early 1997 but declined to varying degrees by late 1997. Twenty-three percent of patients initially and 15% of patients subsequently had emergency department visits that did not lead to hospitalization, 30% initially and 26% subsequently of those who had CD4 cell counts below 0.20 x 10(9)/L did not receive P carinii pneumonia prophylaxis, and 41% initially and 15% subsequently of those who had CD4 cell counts below 0.50 x 10(9)/L did not receive antiretroviral therapy (protease inhibitor or nonnucleoside reverse transcriptase inhibitor). Inferior patterns of care were seen for many of these measures in blacks and Latinos compared with whites, the uninsured and Medicaid-insured compared with the privately insured, women compared with men, and other risk and/or exposure groups compared with men who had sex with men even after CD4 cell count adjustment. With multivariate adjustment, many differences remained statistically significant. Even by early 1998, fewer blacks, women, and uninsured and Medicaid-insured persons had started taking antiretroviral medication (CD4 cell count adjusted P values <.001 to <.005). CONCLUSIONS: Access to care improved from 1996 to 1998 but remained suboptimal. Blacks, Latinos, women, the uninsured, and Medicaid-insured all had less desirable patterns of care. Strategies to ensure optimal care for patients with HIV requires identifying the causes of deficiency and addressing these important shortcomings in care. PMID- 10386556 TI - Novel hMLH1 and hMSH2 germline mutations in African Americans with colorectal cancer. AB - CONTEXT: Germline mutations of the DNA mismatch repair (MMR) genes hMLH1 and hMSH2 have been shown to cosegregate with the colorectal cancer phenotype in multiple hereditary nonpolyposis colorectal cancer (HNPCC) pedigrees. However, the frequency of these mutations among African American patients with colorectal cancer is unknown. OBJECTIVE: To investigate the frequency of germline alterations of the DNA MMR genes hMLH1 and hMSH2 among African Americans affected by HNPCC and early-age onset colorectal cancer. DESIGN, SETTING, AND PATIENTS: Forty unrelated African American HNPCC and early-age onset colorectal cancer patients (8 women, 3 men) were identified from the cancer registry at a National Cancer Institute-designated referral center, 11 of whom were available for and agreed to study participation from January 1997 to February 1998. The mean age of the subjects was 44 years. An additional 50 age- and sex-matched African Americans without personal or family history of colorectal, endometrial, ovarian, urinary tract, or upper gastrointestinal tract malignancy were also studied as a polymorphism control population. In all subjects, genomic DNA was amplified by polymerase chain reaction for all hMLH1 and hMSH2 exons and screened using single strand conformation polymorphism (SSCP) analysis. Samples demonstrating significant SSCP shifts underwent automated nucleotide sequencing analysis. MAIN OUTCOME MEASURE: Frequency of hMLH1 and hMSH2 germline alterations in the affected and control subjects. RESULTS: Germline hMLH1 and hMSH2 mutations were detected in 3 (27%) of the African American colorectal cancer probands studied. Each mutation was novel. Two hMLH1 (an A-->T transversion at codon 26 and a GG- >AT substitution across codons 177 and 178) mutations and 1 hMSH2 mutation (a C- >T transition at codon 389) were identified in 3 female study subjects. Six other hMLH1 and hMSH2 alterations were detected but were presumed to be polymorphisms. Neither missense mutation (at codons 26 and 389) was detected in the control population. CONCLUSIONS: The results of our analysis support an association between the 3 mutations reported and predisposition to colorectal cancer. Further studies are needed to define DNA MMR gene-associated colorectal cancer in African Americans, an understudied population at increased risk of fatal colorectal cancer. PMID- 10386557 TI - A 75-year-old man with congestive heart failure. PMID- 10386558 TI - A 29-year-old man with multiple sclerosis, 1 year later. PMID- 10386559 TI - Risk of transmission of bovine spongiform encephalopathy to humans in the United States: report of the Council on Scientific Affairs. American Medical Association. AB - CONTEXT: The risk of possible transmission of bovine spongiform encephalopathy (BSE) in the United States is a substantial public health concern. OBJECTIVE: To systematically review the current scientific literature and discuss legislation and regulations that have been implemented to prevent the disease. METHODS: Literature review using the MEDLINE, EMBASE, and Lexis/Nexis databases for 1975 through 1997 on the terms bovine spongiform encephalopathy, prion diseases, prions, and Creutzfeldt-Jakob syndrome. The Internet was used to identify regulatory actions and health surveillance. DATA EXTRACTION: MEDLINE, EMBASE, and Lexis/Nexis databases were searched from 1975 through 1997 for English-language articles that provided information on assessment of transmission risk. RESULTS: Unique circumstances in the United Kingdom caused the emergence and propagation of BSE in cattle, including widespread use of meat and bonemeal cattle feed derived from scrapie-infected sheep and the adoption of a new type of processing that did not reduce the amount of infectious prions prior to feeding. Many of these circumstances do not exist in the United States. In the United Kingdom, new variant Creutzfeldt-Jakob disease probably resulted from the ingestion of BSE contaminated processed beef. The United Kingdom and the European Union now have strong regulations in place to stop the spread of BSE. While BSE has not been observed in the United States, the US government has surveillance and response plans in effect. CONCLUSIONS: Current risk of transmission of BSE in the United States is minimal because (1) BSE has not been shown to exist in this country; (2) adequate regulations exist to prevent entry of foreign sources of BSE into the United States; (3) adequate regulations exist to prevent undetected cases of BSE from uncontrolled amplification within the US cattle population; and (4) adequate preventive guidelines exist to prevent high-risk bovine materials from contaminating products intended for human consumption. PMID- 10386560 TI - Reducing blood lead levels: benefits and strategies. PMID- 10386561 TI - Hypoxia is the cause of brain damage in hyponatremia. PMID- 10386562 TI - Independence, governance, and trust: redefining the relationship between JAMA and the AMA. PMID- 10386563 TI - JAMA Patient Page: lead poisoning. PMID- 10386564 TI - Comparing outcomes: comparing systems. PMID- 10386565 TI - Increasing use of Medicare services by veterans with acute myocardial infarction. AB - OBJECTIVES: Some of the nation's 26 million veterans have two government-financed health care entitlements: Medicare and the Department of Veterans Affairs (VA). The aims of this investigation were to examine trends where Medicare-eligible VA users are initially hospitalized for acute myocardial infarction (AMI) and then to assess rates of cardiac procedure use and mortality for veterans initially admitted to each system of care. METHODS: We used VA and HCFA national databases to identify VA users (age range, > or = 65 years) who were initially admitted to a VAMC or Medicare financed hospital (Medicare hospital) with a primary diagnosis of AMI between January 1, 1992, and December 31, 1995, (n = 47,598). We examined the use of cardiac procedures (cardiac catheterization [CC], coronary artery bypass surgery [CABG], and coronary angioplasty [CA] and mortality (30-day and 1 year) by the type of initial admitting hospital within each system of care. RESULTS: Almost 70% of VA users hospitalized for AMI were initially admitted to Medicare hospitals versus VAMCs between 1992 (64%) and 1995 (72%). After adjusting for patient characteristics in logistic models, VA users initially hospitalized in Medicare hospitals were significantly more likely to undergo cardiac procedures than were VA users hospitalized in VAMCs. Differences in the odds of receiving a procedure were most significant when comparing Medicare hospitals with on-site cardiac technology to VA hospitals without on-site cardiac technology (CC: OR 4.34, 95% CI 3.98-4.73; CABG: OR 2.16, 95% CI 1.92-2.43; CA: OR 4.56, 95% CI 3.98-5.25). We found no significant differences in 30-day and 1 year adjusted mortality rates between VA users initially admitted to VAMCs or Medicare hospitals. CONCLUSIONS: Medicare-eligible VA users are increasingly hospitalized in Medicare hospitals for AMI. VA users cared for in Medicare hospitals receive more cardiac procedures but have the same survival as VA users cared for in VAMCs. These findings have policy implications for access, quality, and costs in both systems of care. PMID- 10386566 TI - Satisfaction with care among patients with diabetes in two public health care systems. AB - OBJECTIVES: We compared patient satisfaction among adults with diabetes treated in a Veterans Affairs (VA) health care system with the satisfaction of patients treated in a county-funded health care system. We also examined whether satisfaction differences reflected differences in the process of patient care. DESIGN: Cross-sectional telephone survey of patients recruited from outpatient clinics. SUBJECTS: Five hundred and thirty eight adults, including 310 patients from 4 VA clinics and 228 patients from 2 county clinics. MEASURES: Overall satisfaction with care and satisfaction with 6 separate dimensions of care were measured using the Employee Health Care Value Survey. RESULTS: VA patients were more satisfied than were county patients overall and with 5 of 6 dimensions of their care. These differences increased when we adjusted for patients' sociodemographic and clinical characteristics. VA patients reported more diabetes counseling and shorter waiting times to see their doctor. Each of these self reported process measures was positively correlated with satisfaction and, when taken into account, reduced the differences in satisfaction between the two systems. However, even when we controlled both for patient characteristics and the process of care, VA patients still were more satisfied than were county patients with their care overall as well as with their access to care, communication with providers, and the quality of their health outcomes. CONCLUSIONS: In this study, VA patients with diabetes were more satisfied with their health care than were county patients. Perceived diabetes-related counseling and shorter waiting times contributed to differences between the systems in patient satisfaction but did not explain them completely. PMID- 10386567 TI - The relationship between continuity of care and the health behaviors of patients: does having a usual physician make a difference? AB - BACKGROUND: Implicit in "any willing provider" and "freedom of choice" legislation is the assumption that ongoing provider relationships lead to better patient outcomes on average. Although previous studies have identified associations of usual source of care with medical utilization, its relationship to patient lifestyle has not been examined. OBJECTIVE: To determine the effect of having a usual physician on health behaviors. METHODS: Data on 3,140 adults from the 1995 Mid-Life in the US study were used to estimate logistic regressions of the effect of having a usual physician on exercise, obesity, vitamin-taking, smoking quits, substance abuse behaviors, preventive medical visits, and respondent assessments of the ability to affect one's own health and risk of heart attacks and cancer. RESULTS: Respondents with a usual physician were 3 times as likely to have had a preventive medical visit during the past year. Among lower-income respondents, those with usual physicians were one-half as likely to report substance abuse behaviors. Instrumenting reduced the magnitude of the former but not latter effect. No other significant differences were found. CONCLUSIONS: Strategies designed to foster regular patient-provider relationships may affect certain health behaviors, such as preventive care visits and substance abuse. Yet in the absence of interventions to improve the effectiveness of these relationships, they are unlikely to be a powerful policy instrument for achieving widespread improvements in patient lifestyle choices. PMID- 10386568 TI - The impact of re-engineering and other cost reduction strategies on the staff of a large teaching hospital: a longitudinal study. AB - OBJECTIVES: To examine changes over time in the hospital staff's perceptions of how rapid organizational change, caused by fiscal constraints imposed by governments, affects them, their work environment, and the quality of care and services that they provide. METHODS: A random sample of hospital employees (n = 900) of a large Ontario teaching hospital participated in a longitudinal study which involved surveys at 3 measurement periods over a 2-year period. The questionnaire used in this study included scales reflecting work environment, emotional distress, personal resources, spillover from work to home and vice versa, and perceptions regarding patient care and the hospital as an employer. RESULTS: Significant increases in depression, anxiety, emotional exhaustion, and job insecurity were seen among employees, particularly during the first year of the change process. By the end of the second year, employees reported deterioration in team work, increased unclarity of role, and increased use of distraction to cope. Job demands increased throughout the period whereas little change occurred in the employee's job influence or decision latitude. Overall, the work environment was negatively affected. Although patient care was unaffected in the first year, a significant decline in perceptions of patient care, attention to quality improvement, and overall quality of care were later seen. CONCLUSIONS: This study raises questions about whether hospital re engineering and mergers will be able to achieve the cost reductions sought without sacrificing quality of work life. Along with the rapid change, there was increase in emotional distress among staff and a deterioration in their relationship with their employer. PMID- 10386569 TI - Stability of nursing home quality indicators over time. AB - BACKGROUND: Nursing home quality indicators (QIs) provide a way to support quality assurance and improvement activities and to help ensure that cost savings are based on increased efficiency and not on decreased quality of care. OBJECTIVES: QIs values are expected to change over time. However, to be good indicators of quality, they should be reasonably stable over "short" periods. This paper discusses theoretical and measurement issues affecting stability and examines the stability of QIs over each of two 3-month periods and one 6-month period. SUBJECTS: The study sample included 512 nursing facilities from two states, Kansas and South Dakota. QIs were measured for the first 3 quarters of 1996. MEASURES: Facility level QIs were constructed using three different metrics that each provide a unique perspective of facility performance as follows: the proportion of residents in the facility with the QI condition; the facility's percentile rank in its state; and a variable indicating whether the facility's rank exceeded the 90th percentile in its state. QI stability was assessed using Pearson correlation coefficients, Spearman rank order correlation coefficients, and Cohen's Kappa, as appropriate for the metric. RESULTS: Results indicated high levels of stability for most QIs, with lower levels of stability found to be in keeping with theoretical and measurement considerations. CONCLUSIONS: QIs are reasonably stable over short periods of time. Quality improvement efforts may best be focused on facilities that are consistently poor performers over time, and those that show a large decrease in quality from one quarter to the next. PMID- 10386570 TI - Assessing the performance of utility techniques in the absence of a gold standard. AB - BACKGROUND: Utility techniques are the most commonly used means to assess patient preferences for health outcomes. However, whether utility techniques produce valid measures of preference has been difficult to determine in the absence of a gold standard. OBJECTIVE: To introduce and demonstrate two methods that can be used to evaluate how well utility techniques measure patients' preferences. SUBJECTS AND DESIGN: Patients treated for advanced prostate cancer (n = 57) first ranked eight health states in order of preference. Four utility techniques were then used to elicit patients' utilities for each health state. MEASURES: The rating scale, standard gamble, time trade-off, and a modified version of willingness-to-pay techniques were used to elicit patients' utilities. Technique performance was assessed by computing a differentiation and inconsistency score for each technique. RESULTS: Differentiation scores indicated the rating scale permitted respondents to assign unique utility values to about 70% of the health states that should have received unique values. When the other techniques were used, about 40% or less of the health states that should have received unique utility scores actually did receive unique utility scores. Inconsistency scores, which indicate how often participants assign utility scores that contradict how they value health states, indicated that the willingness-to-pay technique produced the lowest rate of inconsistency (10%). However, this technique did not differ significantly from the rating scale or standard gamble on this dimension. CONCLUSIONS: Differentiation and inconsistency offer a means to evaluate the performance of utility techniques, thereby allowing investigators to determine the extent to which utilities they have elicited for a given decision problem are valid. In the current investigation, the differentiation and inconsistency methods indicated that all four techniques performed at sub-optimal levels, though the rating scale out-performed the standard gamble, time trade-off, and willingness-to-pay techniques. PMID- 10386571 TI - Physician experiences with, and ratings of, managed care organizations in Massachusetts. AB - BACKGROUND: Physicians can provide important information about how managed care plans affect the delivery of health care. Assessments of the quality of managed care plans have rarely used physician evaluations. OBJECTIVES: To elicit physician evaluations of managed care plans and to determine factors associated with those evaluations. RESEARCH DESIGN: Physicians were asked in a mail survey to evaluate a managed care plan they were associated with. SUBJECTS: Probability sample of 1,336 physicians associated with the five largest managed care health plans in Massachusetts. MEASURES: Physicians were asked about the extent to which the management strategies used by a plan influenced their clinical behavior and about the quality of care available to their patients. RESULTS: Evaluations of the plans were significantly different among the eight units evaluated. Some differences between divisions within plans were as large as differences among plans. Physicians reported that the use of education and peer influence influenced their clinical behavior and facilitated the provision of high quality care more than did rules and regulations or financial incentives. Physicians evaluated most positively plans, which they said used educational strategies more than other plans and which used rules and regulations and financial incentives less. Physicians tended to rate staff and group model plans more positively than did other plans. CONCLUSIONS: Physicians can provide important information about the extent to which the organization and operation of managed care plans affect the provision of high quality care. PMID- 10386572 TI - A comprehensive prognostic index to predict survival based on multiple comorbidities: a focus on breast cancer. AB - BACKGROUND: The presence of comorbidities often influences clinical decision making, although many studies exclude patients with comorbid disease for the sake of analysis. OBJECTIVES: The purpose of this study was to develop a Comprehensive Prognostic Index (CPI), designed specifically for breast cancer patients. RESEARCH DESIGN: This study linked Medicare claims with the Kentucky Cancer Registry and developed two models based on 1 year survival; one focused on deaths caused by breast cancer and the other on deaths from all causes. Comorbidities were derived from inpatient and ambulatory claims for up to 2 years before the diagnosis of breast cancer. SUBJECTS: Subjects included a cohort of 848 elderly women first diagnosed with breast cancer in the state of Kentucky in 1993. MEASURES: Each model identified the comorbidities specific to breast cancer that were detrimental to survival, and generated a refined comorbidity index. The CPI integrated these measures with age and stage of cancer into a comprehensive prognostic index. RESULTS: Nearly two-thirds of the patients had evidence of at least one comorbidity. Survival rates decreased with age, more advanced stage, and increased comorbidity burden, as expected. The interaction of comorbidity burden with either age or stage was particularly strong for the older and more advanced stage of cancer. CONCLUSIONS: The CPI could be a useful tool in breast cancer intervention studies and a prognostic aid for clinicians. PMID- 10386573 TI - An analysis of smoking patterns among older adults. AB - BACKGROUND, SUBJECTS, AND METHODS: The 1990 Health Promotion and Disease Prevention Supplement to The National Health Interview Survey was used to develop point-prevalence data about smoking for four age groups, 55 to 64, 65 to 74, 75 to 84, and over 84 and to assess the association of sociodemographics, health status, and health beliefs with a respondent's smoking profile. RESEARCH DESIGN: Chi-square and Cohran-Mantel-Haenszel tests were used to investigate prevalence patterns. Odds ratios generated from logistic regressions were used to indicate degree of association. RESULTS: Fifty-three percent of individuals above the age of 54 smoked in the past and 17% smoked in 1990. Among these smokers, 61% tried to quit and 36% noted that their physicians never advised them to quit. Significant age group differences were noted on the various measures of smoking prevalence. Beliefs about the adverse health effects of smoking were associated with a greater likelihood of never smoking, and among smokers, a greater likelihood of being a former smoker. CONCLUSIONS: Analyses of health behaviors among older adults must recognize the diversity within this age group, and measures of health beliefs should be included in subsequent studies of health behaviors among older adults. Physicians must also play a greater role in discussing smoking with their patients and advocating smoking cessation. PMID- 10386574 TI - Action of thyroid hormones at the cellular level: the mitochondrial target. AB - Thyroid hormones exert profound effects on the energy metabolism. An inspection of the early and more recent literature shows that several targets at the cellular level have been identified. Since their effects on the nuclear signalling pathway have already been well-defined and extensively reviewed, this article focuses on the regulation of mitochondrial activity by thyroid hormones. Mitochondria, by virtue of their biochemical functions, are a natural candidate as a direct target for the calorigenic effects of thyroid hormones. To judge from results coming from various laboratories, it is quite conceivable that mitochondrial activities are regulated both directly and indirectly. Not only triiodo-L-thyronine, but also diiodothyronines are active in regulating the energy metabolism. They influence the resting metabolism in rats with 3,5-diiodo L-thyronine seeming to show a clearer effect. PMID- 10386575 TI - H+-proton-pumping inorganic pyrophosphatase: a tightly membrane-bound family. AB - The earliest known H+-proton-pumping inorganic pyrophosphatase, the integrally membrane-bound H+-proton-pumping inorganic pyrophosphate synthase from Rhodospirillum rubrum, is still the only alternative to H+-ATP synthase in biological electron transport phosphorylation. Cloning of several higher plant vacuolar H+-proton-pumping inorganic pyrophosphatase genes has led to the recognition that the corresponding proteins form a family of extremely similar proton-pumping enzymes. The bacterial H+-proton-pumping inorganic pyrophosphate synthase and two algal vacuolar H+-proton-pumping inorganic pyrophosphatases are homologous with this family, as deduced from their cloned genes. The prokaryotic and algal homologues differ more than the H+-proton-pumping inorganic pyrophosphatases from higher plants, facilitating recognition of functionally significant entities. Primary structures of H+-proton-pumping inorganic pyrophosphatases are reviewed and compared with H+-ATPases and soluble proton pumping inorganic pyrophosphatases. PMID- 10386576 TI - X-ray structural analysis of compensating mutations at the barnase-barstar interface. AB - The crystal structure of the barstar mutants (Y29P) and (Y29D, Y30W) as well as that of the complexes of barstar(Y29P) with wild-type barnase and barnase(H102K) have been determined. These barstar mutants compensate for the dramatic loss of barnase-barstar interaction energy caused by a single mutation of the barnase active site His-102 to a lysine. The latter introduces an uncompensated charge in the pocket at the surface of barstar where Lys-102 is located. The analysis of the structures suggests a mechanism for this compensation based on the solvation of the charge of Lys-102. Additional compensation occurs through the formation of a hydrogen bond. PMID- 10386577 TI - Adaptive hypoxic tolerance in the subterranean mole rat Spalax ehrenbergi: the role of vascular endothelial growth factor. AB - Spalax ehrenbergi has evolved adaptations that allow it to survive and carry out normal activities in a highly hypoxic environment. A key component of this adaptation is a higher capillary density in some Spalax tissues resulting in a shorter diffusion distance for oxygen. Vascular endothelial growth factor (VEGF) is an angiogenic factor that is critical for angiogenesis during development and in response to tissue ischemia. We demonstrate here that VEGF expression is markedly increased in those Spalax tissues with a higher capillary density relative to the normal laboratory rat Rattus norvegicus. Upregulation of VEGF thus appears to be an additional mechanism by which Spalax has adapted to its hypoxic environment. PMID- 10386578 TI - Surfactant protein A and D expression in the porcine Eustachian tube. AB - Surfactant proteins A and D are collectins which are considered to play an important role in the innate immunity of lungs. Our aim was to investigate whether surfactant protein A or D is expressed in the porcine Eustachian tube originating from the upper airways. Both surfactant proteins A and D were present in the epithelial cells of the Eustachian tube, as shown by strong immunostaining. Using RT-PCR and Northern hybridization, these collectins were detected in the Eustachian tube. The present study is the first report demonstrating surfactant protein gene expression in the Eustachian tube. Surfactant proteins A and D may be important in the antibody-independent protection of the middle ear. PMID- 10386579 TI - Cloning of a cDNA encoding diacylglycerol acyltransferase from Arabidopsis thaliana and its functional expression. AB - Triacylglycerols are the most important storage lipids in most plants and animals. Acyl-CoA:diacylglycerol acyltransferase (EC 2.3.1.20) catalyzes the final step of the pathway of triacylglycerol synthesis and is the only step which is unique to this process. Diacylglycerol acyltransferase is required for the synthesis of storage oil in a wide range of oil-bearing seeds and fruits and in floral structures such as petals, anthers and pollen. We describe the first cloning and functional expression of a cDNA encoding diacylglycerol acyltransferase from a plant. The cDNA, cloned from Arabidopsis thaliana, encodes a 520 amino acid protein with a predicted molecular mass of 59.0 kDa which shares 38% amino acid sequence identity with diacylglycerol acyltransferase from mouse. When expressed in insect cell cultures, the protein catalyzes the synthesis of [14C]triacylglycerol from [14C]diacylglycerol and acyl-CoA. Primer extension analysis revealed that the transcription begins 225 bases before the translation start site, yielding an unusually long 5' untranslated region. The gene is expressed in a wide range of tissues but most strongly in developing embryos and petals of flowers. PMID- 10386580 TI - The phosphoinositide 3-kinase/Akt pathway is activated by daunorubicin in human acute myeloid leukemia cell lines. AB - Daunorubicin induces apoptosis in myeloid leukemia cells by activation of neutral sphingomyelinase and ceramide generation occurring 4-10 min after daunorubicin addition. We show here that daunorubicin is able to increase the phosphoinositide 3-kinase activity and enhance intracellular phosphoinositide 3-kinase lipid products prior to ceramide generation. Daunorubicin activates Akt, a downstream phosphoinositide 3-kinase effector. Interestingly, the phosphoinositide 3-kinase inhibitors wortmannin and LY294002 accelerate daunorubicin-induced apoptosis in U937 cells. The phosphoinositide 3-kinase/Akt pathway has been involved in cell survival following serum deprivation, tumor necrosis factor alpha, anti-Fas and UV radiations. Our results suggest that anti-tumor agents such as daunorubicin may also activate anti-apoptotic signals that could contribute to drug resistance. PMID- 10386581 TI - Domain IV of elongation factor G from Thermus thermophilus is strictly required for translocation. AB - Two truncated variants of elongation factor G from Thermus thermophilus with deletion of its domain IV have been constructed and the mutated genes were expressed in Escherichia coli. The truncated factors were produced in a soluble form and retained a high thermostability. It was demonstrated that mutated factors possessed (1) a reduced affinity to the ribosomes with an uncleavable GTP analog and (2) a specific ribosome-dependent GTPase activity. At the same time, in contrast to the wild-type elongation factor G, they were incapable to promote translocation. The conclusions are drawn that (1) domain IV is not involved in the GTPase activity of elongation factor G, (2) it contributes to the binding of elongation factor G with the ribosome and (3) is strictly required for translocation. These results suggest that domain IV might be directly involved in translocation and GTPase activity of the factor is not directly coupled with translocation. PMID- 10386582 TI - 3-Hydroxy-3-methylglutaryl CoA reductase inhibitors reduce serum triglyceride levels through modulation of apolipoprotein C-III and lipoprotein lipase. AB - Statins are hypolipidemic drugs which not only improve cholesterol but also triglyceride levels. Whereas their cholesterol-reducing effect involves inhibition of de novo biosynthesis of cellular cholesterol through competitive inhibition of its rate-limiting enzyme 3-hydroxy-3-methylglutaryl CoA reductase, the mechanism by which they lower triglycerides remains unknown and forms the subject of the current study. Treatment of normal rats for 4 days with simvastatin decreased serum triglycerides significantly, whereas it increased high density lipoprotein cholesterol moderately. The decrease in triglyceride concentrations after simvastatin was caused by a reduction in the amount of very low density lipoprotein particles which were of an unchanged lipid composition. Simvastatin administration increased the lipoprotein lipase mRNA and activity in adipose tissue and heart. This effect on lipoprotein lipase was accompanied by decreased mRNA as well as plasma levels of the lipoprotein lipase inhibitor apolipoprotein C-III. These results suggest that the triglyceride-lowering effect of statins involves a stimulation of lipoprotein lipase-mediated clearance of triglyceride-rich lipoproteins. PMID- 10386583 TI - Isoforms of cytochrome P450 on organic nitrate-derived nitric oxide release in human heart vessels. AB - Glutathione S-transferases and the cytochrome P450 system have been proposed for the vascular biotransformation systems in the metabolic activation of organic nitrates. The present study was designed to elucidate the role of human cytochrome P450 isoforms on nitric oxide formation from organic nitrates using lymphoblast microsomes transfected with human CYP isoforms cDNA. CYP3A4 transfected microsomes had the most effective potential of nitric oxide formation from isosorbide dinitrate. Anti-CYP3A2 antibody (which cross-reacts with CYP3A4) or ketoconazole (an inhibitor of the CYP3A superfamily) inhibited nitric oxide formation from isosorbide dinitrate in rat heart microsomes. Immunohistochemistry of human heart also showed intense bindings of CYP3A4 antibody in the endothelium of the endocardium and coronary vessels. These results suggest that the CYP3A4 NADPH-cytochrome P450 reductase system specifically participates in nitric oxide formation from isosorbide dinitrate. PMID- 10386584 TI - Cloning and characterization of hGMEB1, a novel glucocorticoid modulatory element binding protein. AB - A 21-bp element called glucocorticoid modulatory element (GME) modulates the glucocorticoid receptor-mediated responses via the binding of an as yet poorly characterized transacting complex of proteins containing the 88-kDa GMEB1 and the 67-kDa GMEB2. Using heat shock protein 27 (HSP27) as bait in the yeast two-hybrid assay, we cloned a 1.83-kb cDNA encoding a novel 573-amino acid protein called human GMEB1 (hGMEB1). hGMEB1 possesses a KDWK domain, contains sequences almost identical (36/38) to three tryptic peptides of rat GMEB1 and shares 38% identity with rat GMEB2. hGMEB1 is ubiquitously expressed as a 85-kDa protein in all cell lines and tissues examined. In vitro translated hGMEB1 bound specifically to GME oligonucleotides yielding a complex of similar size to the complex obtained using rat liver nuclear extracts. Both complexes were supershifted with an antibody specific to hGMEB1. Co-immunoprecipitation experiments confirmed the in vivo interaction of HSP27 with hGMEB1. PMID- 10386585 TI - Localization of the human caveolin-3 gene to the D3S18/D3S4163/D3S4539 locus (3p25), in close proximity to the human oxytocin receptor gene. Identification of the caveolin-3 gene as a candidate for deletion in 3p-syndrome. AB - Caveolin-3, a muscle-specific caveolin-related protein, is the principal structural protein of caveolae membrane domains in striated muscle cell types (cardiac and skeletal). Recently, we identified an autosomal dominant form of limb girdle muscular dystrophy in humans that is due to mutations within exon 2 of the caveolin-3 gene (3p25). However, the detailed location of the human caveolin-3 gene and its position with regard to neighboring genes remains unknown. Here, we have isolated three independent BAC clones containing the human caveolin-3 gene. Using a PCR-based approach, we determined that these clones contain both exons 1 and 2 of the human caveolin-3 gene. In addition, we performed microsatellite marker analysis of these BAC clones, using a panel of 13 markers that are known to map within the 3p25 region. Our results indicate that these BAC clones contain the following three markers: D3S18, SHGC-1079 (also known as D3S4163) and D3S4539. Interestingly, D3S18 is a marker for two known human diseases, von Hippel-Lindau disease and 3p-syndrome. As D3S4163 and D3S4539 are known to map in the vicinity of the 3' end of the human oxytocin receptor gene, we determined if these caveolin-3 positive BACs also contain the oxytocin receptor gene. We show that (i) these BACs contain all four exons of the oxytocin receptor gene and (ii) that the genes encoding caveolin-3 and the oxytocin receptor are located approximately 7-10 kb apart and in the opposite orientation. As 3p-syndrome is characterized by cardiac septal defects and caveolin-3 is expressed primarily in the heart and skeletal muscle, caveolin-3 is a candidate gene that may be deleted in 3p-syndrome. PMID- 10386586 TI - A novel anti-hypertensive peptide derived from ovalbumin induces nitric oxide mediated vasorelaxation in an isolated SHR mesenteric artery. AB - In this report, we deal with the isolation of a novel vasorelaxing peptide from a chymotryptic digest of ovalbumin and its vasorelaxing activities. This peptide is composed of Arg-Ala-Asp-His-Pro-Phe (RADHPF) in its sequence, corresponding to residues 359-364 of ovalbumin. This peptide (30-300 microM) exerted a dose dependent vasodilation in an isolated mesenteric artery from a spontaneously hypertensive rat which was pre-constricted by phenylephrine, besides the relaxation being endothelium-dependent. It is noteworthy that the nitric oxide synthase inhibitor N(G)-nitro-L-arginine methyl ester inhibited this relaxation, implying involvement of nitric oxide in its mechanism of action. Following oral administration of RADHPF at a dose of 10 mg/kg, the systolic blood pressure in a spontaneously hypertensive rat was significantly lowered. PMID- 10386587 TI - Demonstration of a novel sulfotransferase in fetal bovine serum, which transfers sulfate to the C6 position of the GalNAc residue in the sequence iduronic acid alpha1-3GalNAc beta1-4iduronic acid in dermatan sulfate. AB - A novel sulfotransferase activity was discovered in fetal bovine serum using pig skin dermatan sulfate as an acceptor and [35S]3'-phosphoadenosine 5' phosphosulfate as a sulfate donor. The enzyme was separated from chondroitin:GalNAc 6-O-sulfotransferase by chromatographic techniques. Enzymatic analysis of the reaction products demonstrated that the enzyme transferred sulfate to the C6 position of the GalNAc residue in the sequence -iduronic acid alpha1-3GalNAc beta1-4iduronic acid-. Thus, the enzyme has been identified as a hitherto unreported dermatan sulfate:GalNAc 6-O-sulfotransferase. The finding is in sharp contrast to the current concept that in dermatan sulfate biosynthesis GalNAc 4-O-sulfation is a prerequisite for iduronic acid formation by C5 epimerase. PMID- 10386588 TI - Probing the reactivity of nucleophile residues in human 2,3 diphosphoglycerate/deoxy-hemoglobin complex by aspecific chemical modifications. AB - The use of aspecific methylation reaction in combination with MS procedures has been employed for the characterization of the nucleophilic residues present on the molecular surface of the human 2,3-diphosphoglycerate/deoxy-hemoglobin complex. In particular, direct molecular weight determinations by ESMS allowed to control the reaction conditions, limiting the number of methyl groups introduced in the modified globin chains. A combined LCESMS-Edman degradation approach for the analysis of the tryptic peptide mixtures yielded to the exact identification of methylation sites together with the quantitative estimation of their degree of modification. The reactivities observed were directly correlated with the pKa and the relative surface accessibility of the nucleophilic residues, calculated from the X-ray crystallographic structure of the protein. The results here described indicate that this methodology can be efficiently used in aspecific modification experiments directed to the molecular characterization of the surface topology in proteins and protein complexes. PMID- 10386589 TI - Fibrillin degradation by matrix metalloproteinases: identification of amino- and carboxy-terminal cleavage sites. AB - Fibrillin molecules form the structural framework of elastic fibrillin-rich microfibrils of the extracellular matrix. We have investigated the proteolysis of recombinant fibrillin molecules by five matrix metalloproteinases. Cleavage sites were defined at the carboxy-terminal end of the fibrillin-1 proline-rich region and the corresponding fibrillin-2 glycine-rich region (exon 10), and within exon 49 towards the carboxy-terminus of fibrillin-1. Cleavage at these sites is predicted to disrupt the structure and function of the fibrillin-rich microfibrils. PMID- 10386590 TI - Identification of RPE65 in transformed kidney cells. AB - The protein RPE65 has an important role in retinoid processing and/or retinoid transport in the eye. Retinoids are involved in cell differentiation, embryogenesis and carcinogenesis. Since the kidney is known as an important site for retinoid metabolism, the expression of RPE65 in normal kidney and transformed kidney cells has been examined. The RPE65 mRNA was detected in transformed kidney cell lines including the human embryonic kidney cell line HEK293 and the African green monkey kidney cell lines COS-1 and COS-7 by reverse transcription PCR. In contrast, it was not detected in human primary kidney cells or monkey kidney tissues under the same PCR conditions. The RPE65 protein was also identified in COS-7 and HEK293 cells by Western blot analysis using a monoclonal antibody to RPE65, but not in the primary kidney cells or kidney tissues. The RPE65 cDNA containing the full-length encoding region was amplified from HEK293 and COS-7 cells. DNA sequencing showed that the RPE65 cDNA from HEK293 cells is identical to the RPE65 cDNA from the human retinal pigment epithelium. The RPE65 from COS-7 cells shares 98 and 99% sequence identity with human RPE65 at the nucleotide and amino acid levels, respectively. Moreover, the RPE65 mRNA was detected in three out of four renal tumor cultures analyzed including congenital mesoblastic nephroma and clear cell sarcoma of the kidney. These results demonstrated that transformed kidney cells express this retinoid processing protein, suggesting that these transformed cells may have an alternative retinoid metabolism not present in normal kidney cells. PMID- 10386591 TI - Contribution of separate tryptophan residues to intrinsic fluorescence of actin. Analysis of 3D structure. AB - The location of tryptophan residues in the actin macromolecule was studied on the basis of the known 3D structure. For every tryptophan residue the polarity and packing density of their microenvironments were evaluated. To estimate the accessibility of the tryptophan residues to the solvent molecules it was proposed to analyze the radial dependence of the packing density of atoms in the macromolecule about the geometric center of the indole rings of the tryptophan residues. The proposed analysis revealed that the microenvironment of tryptophan residues Trp-340 and Trp-356 has a very high density. So these residues can be regarded as internal and inaccessible to solvent molecules. Their microenvironment is mainly formed by non-polar groups of protein. Though the packing density of the Trp-86 microenvironment is lower, this tryptophan residue is apparently also inaccessible to solvent molecules, as it is located in the inner region of macromolecule. Tryptophan residue Trp-79 is external and accessible to the solvent. All residues that can affect tryptophan fluorescence were revealed. It was found that in the close vicinity of tryptophan residues Trp 79 and Trp-86 there are a number of sulfur atoms of cysteine and methionine residues that are known to be effective quenchers of tryptophan fluorescence. The most essential is the location of SG atom of Cys-10 near the NE1 atom of the indole ring of tryptophan residue Trp-86. On the basis of microenvironment analysis of these tryptophan residues and the evaluation of energy transfer between them it was concluded that the contribution of tryptophan residues Trp-79 and Trp-86 must be low. Intrinsic fluorescence of actin must be mainly determined by two other tryptophan residues--Trp-340 and Trp-356. It is possible that the unstrained conformation of tryptophan residue Trp-340 and the existence of aromatic rings of tyrosine and phenylalanine and proline residues in the microenvironments of tryptophan residues Trp-340 and Trp-356 are also essential to their blue fluorescence spectrum. PMID- 10386592 TI - Mistletoe lectin dissociates into catalytic and binding subunits before translocation across the membrane to the cytoplasm. AB - Hybridomas producing monoclonal antibodies (mAbs) against the mistletoe lectin A chain (MLA) were obtained to investigate the intracellular routing and translocation of ribosome-inactivating proteins. Anti-MLA mAb MNA5 did not bind the holotoxin but interacted with isolated MLA. This epitope was not recognized upon MLA denaturation or conjugation of MLA with the ricin binding subunit (RTB). Furthermore, the mAbs did not appreciably react with a panel of MLA synthetic octapeptides linked to the surface of polyethylene pins. A study of the cytotoxicity of mistletoe lectin, ricin, and chimeric toxin MLA/RTB for the hybridomas revealed that interchain disulfide bond reduction and subunit dissociation are required for cytotoxic activity of mistletoe lectin. PMID- 10386593 TI - Chromatin structural transitions in Drosophila embryo cell-free extract result in a high conformational flexibility of nucleosomal DNA. AB - DNA within chromatin has considerably more restricted flexibility in comparison with naked DNA. This raises the main question of how the functioning multi-enzyme complexes overcome the nucleosomal level of DNA packaging. We studied the DNA conformational flexibility of reconstituted chromatin in a cell-free system derived from Drosophila embryo extracts. Using this system, we have found evidence for a energy-independent chromatin remodelling process that efficiently destabilizes the nucleosome structure resulting in a high conformational flexibility of nucleosomal DNA. The described chromatin remodelling process may lay on the basis of defined molecular principles governing the molecular heterogeneity of chromatin structures in vivo. PMID- 10386594 TI - Mildly oxidized GAPDH: the coupling of the dehydrogenase and acyl phosphatase activities. AB - The hydrogen peroxide-induced 'non-phosphorylating' activity of D-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is shown to be a result of the successive action of two forms of the enzyme subunits: one catalyzing production of 1,3 bisphosphoglycerate, and the other performing its hydrolytic decomposition. The latter form is produced by mild oxidation of GAPDH in the presence of a low hydrogen peroxide concentration when essential Cys-149 is oxidized to the sulfenate derivative. The results obtained with a C153S mutant of Bacillus stearothermophilus GAPDH rule out the possibility that intrasubunit acyl transfer between Cys-149 and a sulfenic form of Cys-153 is required for the 'non phosphorylating' activity of the enzyme. PMID- 10386595 TI - The structure of chromatophores from purple photosynthetic bacteria fused with lipid-impregnated collodion films determined by near-field scanning optical microscopy. AB - Lipid-impregnated collodion (nitrocellulose) films have been frequently used as a fusion substrate in the measurement and analysis of electrogenic activity in biological membranes and proteoliposomes. While the method of fusion of biological membranes or proteoliposomes with such films has found a wide application, little is known about the structures formed after the fusion. Yet, knowledge of this structure is important for the interpretation of the measured electric potential. To characterize structures formed after fusion of membrane vesicles (chromatophores) from the purple bacterium Rhodobacter sphaeroides with lipid-impregnated collodion films, we used near-field scanning optical microscopy. It is shown here that structures formed from chromatophores on the collodion film can be distinguished from the lipid-impregnated background by measuring the fluorescence originating either from endogenous fluorophores of the chromatophores or from fluorescent dyes trapped inside the chromatophores. The structures formed after fusion of chromatophores to the collodion film look like isolated (or sometimes aggregated, depending on the conditions) blisters, with diameters ranging from 0.3 to 10 microm (average approximately 1 microm) and heights from 0.01 to 1 microm (average approximately 0.03 microm). These large sizes indicate that the blisters are formed by the fusion of many chromatophores. Results with dyes trapped inside chromatophores reveal that chromatophores fused with lipid-impregnated films retain a distinct internal water phase. PMID- 10386596 TI - The novel transmembrane Escherichia coli proteins involved in the amino acid efflux. AB - A novel gene of Escherichia coli, rhtB, has been characterized. Amplification of this gene provides resistance to homoserine and homoserine lactone. Another E. coli gene, rhtC, provides resistance to threonine. The homologues of RhtB are widely distributed among various eubacteria and archaea, from one to 12 copies of family members that differ in their primary structure were found in the genomes. Most of them are genes that encode hypothetical transmembrane proteins. Experimental data that indicate participation of the rhtB product in the excretion of homoserine have been obtained. PMID- 10386597 TI - Kinetics of peroxidases in guinea pig bone marrow under immunostimulation. AB - Eosinophil peroxidase and myeloperoxidase play an important role in the host defense. Both enzymes are present in bone marrow, synthesized by blood progenitor cells. This research investigated the kinetic properties of peroxidases under immunostimulation in guinea pig bone marrow. Results suggest that there are at least two myeloperoxidase isozymes and at least three eosinophil peroxidase isozymes in guinea pig bone marrow and that some of these isozymes are expressed upon immunostimulation. PMID- 10386598 TI - Alpha1,3-fucosyltransferase IX (Fuc-TIX) is very highly conserved between human and mouse; molecular cloning, characterization and tissue distribution of human Fuc-TIX. AB - The amino acid sequence of Fuc-TIX is very highly conserved between mouse and human. The number of non-synonymous nucleotide substitutions of the Fuc-TIX gene between human and mouse was strikingly low, and almost equivalent to that of the alpha-actin gene. This indicates that Fuc-TIX is under a strong selective pressure of preservation during evolution. The human Fuc-TIX (hFuc-TIX) showed a unique characteristics, i.e. hFuc-TIX was not activated by Mn2+ and Co2+, whereas hFuc-TIV and hFuc-TVI were activated by the cations. The hFuc-TIX transcripts were abundantly expressed in brain and stomach, and interestingly were detected in spleen and peripheral blood leukocytes. PMID- 10386599 TI - Helicobacter pylori lipopolysaccharide enhances the expression of NADPH oxidase components in cultured guinea pig gastric mucosal cells. AB - Recently, we showed that cultured guinea pig gastric pit cells possess a phagocyte NADPH oxidase-like activity, which was up-regulated by Helicobacter pylori lipopolysaccharide. We demonstrate here that these cells express all of the phagocyte NADPH oxidase components (gp91-, p22-, p67-, p47-, and p40-phoxes). Treatment with lipopolysaccharide increased the expression of gp91-, p22-, and p67-phoxes, but not that of p47- and p40-phoxes. Intriguingly, the p67-phox expression consistently correlated with up-regulation of superoxide anion producing ability. Thus, the gastric pit cell NADPH oxidase may play an important role in regulation of the inflammatory response associated with H. pylori infection. PMID- 10386600 TI - Skp2 induction and phosphorylation is associated with the late G1 phase of proliferating rat hepatocytes. AB - The changes in phosphoproteins purified with the affinity peptide p9CKShs1 were analyzed from extracts of regenerating rat livers in order to define some G1 and G1/S regulations characteristic of mature hepatocytes stimulated to proliferate. We observed a 47 kDa phosphoprotein that occurred first at the end of G1 before peaking in the S phase. P47 was also found to be phosphorylated in late G1 in primary hepatocyte cultures stimulated with mitogens. P47 was still phosphorylated in extracts depleted of Cdc2, but to a lesser extent after Cdk2 depletion. This phosphoprotein was identified as Skp2. (i) P47 shared the same electrophoretic mobility than Skp2, a cell cycle protein essential for S phase entry in human fibroblasts; (ii) Skp2, like P47, started to be expressed and was highly phosphorylated during the G1/S transition of hepatocytes stimulated to proliferate in vivo and in vitro; (iii) P47 was specifically immunoprecipitated by an antibody directed against Skp2. In addition, cyclin A/Cdk2 complexes from regenerating liver clearly interacted with Skp2. This is the first demonstration that Skp2 is induced and phosphorylated in the late G1 and S phase of hepatocytes in vivo in regenerating liver as well as in vitro in mitogen-stimulated hepatocytes. PMID- 10386601 TI - Evidence against the regulation of caldesmon inhibitory activity by p42/p44erk mitogen-activated protein kinase in vitro and demonstration of another caldesmon kinase in intact gizzard smooth muscle. AB - The effect of direct phosphorylation by recombinant p44erk1 mitogen-activated protein kinase on the inhibitory activity of caldesmon and its C-terminal fragment H1 was studied in vitro. Neither inhibition of actin-tropomyosin activated ATPase of heavy meromyosin by caldesmon or H1, nor inhibition of the actin-tropomyosin motility over heavy meromyosin by H1 was significantly affected by the phosphorylation while only a moderate effect on the actin-activated component of heavy meromyosin ATPase inhibition was observed. Phosphopeptide mapping of caldesmon immunoprecipitated from [32P]PO4-labelled intact gizzard strips revealed that it is predominantly phosphorylated at mitogen-activated protein kinase sites in unstimulated tissue and that it is stimulated for 1 h with phorbol 12,13-dibutyrate. We find that phorbol 12,13-dibutyrate also induces a transitory phosphorylation of caldesmon peaking at 15 min after addition and this phosphorylation is not attributed to mitogen-activated protein kinase, protein kinase C, Ca2+/calmodulin-dependent kinase II or casein kinase II. We suggest that a yet unidentified kinase, rather than mitogen-activated protein kinase, may be involved in regulation of the caldesmon function in vivo. PMID- 10386602 TI - Anisoosmotic regulation of the Nopp140 mRNA in H4IIE rat hepatoma cells and primary hepatocytes. AB - Using the differential display polymerase chain reaction osmosensitive regulation of mRNA levels of the nucleolar phosphoprotein of 140 kDa (Nopp140) was found in H4IIE rat hepatoma cells. These levels were downregulated after hypoosmotic exposure in H4IIE cells and primary rat hepatocytes. Hyperosmotic incubation increased Nopp140 mRNA levels in H4IIE cells but not in hepatocytes. Inhibition of p38MAPK or MAP kinase kinase upstream of Erk-1 and Erk-2 decreased Nopp140 mRNA levels but did not prevent their osmosensitivity. Because Nopp140 is involved in the regulation of transcriptional activity it could play a role in the osmosignalling pathway towards gene expression in H4IIE cells and hepatocytes. PMID- 10386603 TI - The jasmonate-induced 60 kDa protein of barley exhibits N-glycosidase activity in vivo. AB - Upon jasmonate treatment barley leaf segments express a putative ribosome inactivating protein (JIP60). The influence of this protein on translation in planta has been analysed by using barley plants and tobacco plants transformed with a barley cDNA encoding JIP60. In both plant systems JIP60 exhibited N glycosidase activity in vivo. The depurination of the 25S rRNA of tobacco and barley ribosomes led to accumulation of translationally inactive polysomes. PMID- 10386604 TI - Atomic force microscopy sees nucleosome positioning and histone H1-induced compaction in reconstituted chromatin. AB - We addressed the question of how nuclear histones and DNA interact and form a nucleosome structure by applying atomic force microscopy to an in vitro reconstituted chromatin system. The molecular images obtained by atomic force microscopy demonstrated that oligonucleosomes reconstituted with purified core histones and DNA yielded a 'beads on a string' structure with each nucleosome trapping 158 +/- 27 bp DNA. When dinucleosomes were assembled on a DNA fragment containing two tandem repeats of the positioning sequence of the Xenopus 5S RNA gene, two nucleosomes were located around each positioning sequence. The spacing of the nucleosomes fluctuated in the absence of salt and the nucleosomes were stabilized around the range of the positioning signals in the presence of 50 mM NaCl. An addition of histone H1 to the system resulted in a tight compaction of the dinucleosomal structure. PMID- 10386605 TI - Biosynthesis of sialylated and fucosylated selectin ligands of HL-60 cells in vitro. Midchain alpha3-fucose units inhibit terminal alpha6-sialylation but not alpha3-sialylation of polylactosamines. AB - Polylactosamines Neu5Ac alpha2-3'Lex beta1-3'Lex beta1-3'Lex and Neu5Ac alpha2 3'LNbeta1-3'Lex beta1-3'Lex [Lex, Gal beta1-4(Fuc alpha1-3)GlcNAc; LN, Gal beta1 4GlcNAc] decorate selectin counterreceptors in human HL-60 cells. Here, we show that HL-60 cell lysates catalyze distal alpha3-sialylation of LNbeta1-3'LNbeta1 3'LN and LNbeta1-3'Lex beta1-3'Lex efficiently, outlining two potential sets of biosynthetic pathways leading to the selectin ligands. In one set, alpha3 sialylation precedes internal fucosylation of the polylactosamine backbone, whereas in the other one, internal fucosylation is initiated before alpha3 sialylation. In contrast to alpha3-sialylation, LNbeta1-3'Lex beta1-3'Lex was alpha6-sialylated much less efficiently than LNbeta1-3'LNbeta1-3'LN by HL-60 cell lysates. Hence, internal fucosylation may regulate the extent of alpha6 sialylation of polylactosamines in these cells. PMID- 10386606 TI - Troglitazone inhibits angiotensin II-induced extracellular signal-regulated kinase 1/2 nuclear translocation and activation in vascular smooth muscle cells. AB - The thiazolidinedione troglitazone inhibits angiotensin II-induced extracellular signal-regulated kinase 1/2 mitogen-activated protein kinase activity in vascular smooth muscle cells. Activation of extracellular signal-regulated kinase 1/2 by angiotensin II is a multistep process involving both its phosphorylation by mitogen-activated protein kinase extracellular signal-regulated kinase kinase in the cytoplasm and a subsequent translocation to the nucleus. The cytoplasmic activation of extracellular signal-regulated kinase 1/2 in vascular smooth muscle cells proceeds through the protein kinase Czeta --> mitogen-activated protein kinase extracellular signal-regulated kinase kinase --> extracellular signal regulated kinase pathway. Troglitazone did not affect the angiotensin II-induced activation of protein kinase Czeta or its downstream signaling kinases extracellular signal-regulated kinase 1/2 in the cytosol. In contrast, angiotensin II-induced activation of protein kinase Czeta and extracellular signal-regulated kinase 1/2 in the nucleus were both inhibited by troglitazone. Nuclear translocation of extracellular signal-regulated kinase 1/2 induced by angiotensin II was completely blocked by troglitazone. Protein kinase Czeta, however, did not translocate upon angiotensin II stimulation. Troglitazone, therefore, inhibits both angiotensin II-induced nuclear translocation of extracellular signal-regulated kinase 1/2 and the nuclear activity of its upstream signaling kinase protein kinase Czeta. Since extracellular signal regulated kinase 1/2 nuclear translocation may be a critical signaling step for multiple growth factors that stimulate vascular smooth muscle cells proliferation and migration, troglitazone may provide a new therapeutical approach for the prevention and treatment of atherosclerosis and restenosis. PMID- 10386607 TI - Predicting the protein folding nucleus from sequences [correction of a sequence]. AB - Understanding the mechanism of protein folding would allow prediction of the three-dimensional structure from sequence data alone. It has been shown that small proteins fold in a small number of kinetic steps and that significantly populated intermediate states exist for some of them. Studies of these intermediates have demonstrated the existence of specific interactions established during the initial stages of folding. Comparison of the amino acids participating in these specific and essential interactions and constituting the folding nucleus with conserved hydrophobic positions of a given fold shows a striking correspondence. This finding opens the perspective of predicting the folding nucleus knowing only a set of divergent sequences of a protein family. PMID- 10386608 TI - Nitric oxide mediates brain mitochondrial maturation immediately after birth. AB - The possible role of nitric oxide (*NO) in brain mitochondrial maturation was studied. Within the first 5 min after birth, a sharp increase in ATP concentrations was observed, coinciding with an increase in mitochondrial complex II-III (succinate-cytochrome c reductase) activity, while complex I (NADH-CoQ1 reductase) and complex IV (cytochrome c oxidase) activities remained unchanged. Under the same circumstances, cGMP concentrations were increased by 5 min after birth, correlating significantly with ATP concentrations. Since ATP concentrations also correlated significantly with mitochondrial complex II-III activity, these three parameters may be associated. Inhibition of *NO synthase activity brought about by the administration of N(omega)-nitro-L-arginine monomethyl ester to mothers prevented the postnatal increase in cGMP and ATP levels and complex II-III activity. These results suggest that early postnatal mitochondrial maturation in the brain is a *NO-mediated process. PMID- 10386609 TI - Hypochlorite modification of high density lipoprotein: effects on cholesterol efflux from J774 macrophages. AB - The present study was aimed at investigating effects of hypochlorite (HOCl) modification of high density lipoproteins subclass 3 (HDL3) on their ability for cellular cholesterol removal from permanent J774 macrophages. Our findings indicate that HOCl (added as reagent or generated enzymatically by the myeloperoxidase/H2O2/Cl- system) damages apolipoprotein A-I, the major protein component of HDL3. Fatty acid analysis of native and HOCl-modified HDL3 revealed that unsaturated fatty acids in both major lipid subclasses (phospholipids and cholesteryl esters) are targets for HOCl attack. HOCl modification resulted in impaired HDL3-mediated cholesterol efflux from J774 cells, regardless of whether reagent or enzymatically generated HOCl was used to modify the lipoprotein. Decreased cholesterol efflux was also observed after HOCl modification of reconstituted HDL particles. Impairment of cholesterol efflux from macrophages was noticed at low and physiologically occurring HOCl concentrations. PMID- 10386610 TI - Thiazolidinedione inhibits production of RANTES in a cytokine-treated human lung epithelial cell line. AB - The chemokine RANTES is a potent chemoattractant for eosinophils. RANTES is produced by lung epithelial cells during eosinophil-rich inflammatory diseases such as asthma. In this study, we examined the effects of thiazolidinediones (TZD) on RANTES expression in a human lung epithelial cell line, A549. In A549 cells, interleukin-1beta and tumor necrosis factor-alpha induced endogenous RANTES protein secretion, mRNA expression, and promoter activity. The TZD inhibited these effects. Our data indicate that the suppression of the expression of RANTES can be accomplished by TZD treatment, raising the possibility that TZD might be of therapeutic value in diseases such as asthma. PMID- 10386611 TI - The Drosophila gene 2A5 complements the defect in mitochondrial F1-ATPase assembly in yeast lacking the molecular chaperone Atp11p. AB - Assembly of mitochondrial F1-ATPase in Saccharomyces cerevisiae requires the molecular chaperone, Atp11p. Database searches have identified protein sequences from Schizosaccharomyces pombe and two species of Drosophila that are homologous to S. cerevisiae Atp11p. A cDNA encoding the putative Atp11p from Drosophila yakuba was shown to complement the respiratory deficient phenotype of yeast harboring an atp11::HIS3 disruption allele. Furthermore, the product of this Drosophila gene was shown to interact with the S. cerevisiae F1 beta subunit in the yeast two-hybrid assay. These results indicate that Atp11p function is conserved in higher eukaryotes. PMID- 10386612 TI - Elafin (elastase-specific inhibitor) has anti-microbial activity against gram positive and gram-negative respiratory pathogens. AB - Elafin (elastase-specific inhibitor) is a low molecular weight inhibitor of neutrophil elastase which is secreted in the lung. Using synthetic peptides corresponding to full-length elafin (H2N-1AVT.....95Q-OH), the NH2-terminal domain (H2N-1AVT.....50K-OH) and the COOH-terminal domain (H2N-51PGS.....95Q-OH), we demonstrate that elafin's anti-elastase activity resides exclusively in the COOH-terminus. Several characteristics of elafin suggest potential anti-microbial activity. The anti-microbial activity of elafin, and of its two structural domains, was tested against the respiratory pathogens Pseudomonas aeruginosa and Staphylococcus aureus. Elafin killed both bacteria efficiently, with 93% killing of P. aeruginosa by 2.5 microM elafin and 48% killing of S. aureus by 25 microM elafin. For both organisms, full-length elafin was required to optimise bacterial killing. These findings represent the first demonstration of co-existent anti proteolytic and anti-microbial functions for elafin. PMID- 10386613 TI - The effects of the selective ROCK inhibitor, Y27632, on ET-1-induced hypertrophic response in neonatal rat cardiac myocytes--possible involvement of Rho/ROCK pathway in cardiac muscle cell hypertrophy. AB - A small GTPase, Rho, participates in agonist-induced cytoskeletal organization and gene expression in many cell types including cardiac myocytes. However, little is known about the functions of Rho's downstream targets in cardiac myocytes. We examined the role of ROCK, a downstream target of Rho, in ET-1 induced hypertrophic response. Y27632, a selective ROCK inhibitor, inhibited ET-1 induced increases in natriuretic peptide production, cell size, protein synthesis, and myofibrillar organization. In addition, a dominant-negative mutant of p160ROCK suppressed ET-1-induced transcription of the BNP gene. These findings suggest that the Rho/ROCK pathway is an important component of ET-1-induced hypertrophic signals in cardiac myocytes. PMID- 10386614 TI - Effect of mutations found in carbohydrate-deficient glycoprotein syndrome type IA on the activity of phosphomannomutase 2. AB - Seven mutant forms of human phosphomannomutase 2 were produced in Escherichia coli and purified. These mutants had a Vmax of 0.2-50% of the wild enzyme and were unstable. The least active protein (R141H) bears a very frequent mutation, which has never been found in the homozygous state whereas the second least active protein (D188G) corresponds to a mutation associated with a particularly severe phenotype. We conclude that total lack of phosphomannomutase 2 is incompatible with life. Another conclusion is that the elevated residual phosphomannomutase activity found in fibroblasts of some patients is contributed by their mutated phosphomannomutase 2. PMID- 10386615 TI - PCR random mutagenesis into Escherichia coli serine acetyltransferase: isolation of the mutant enzymes that cause overproduction of L-cysteine and L-cystine due to the desensitization to feedback inhibition. AB - PCR random mutagenesis in the cysE gene encoding Escherichia coli serine acetyltransferase was employed to isolate the mutant enzymes that, due to a much less feedback inhibition by L-cysteine, cause overproduction of L-cysteine and L cystine in the recombinant strains. The L-cysteine auxotrophic and non-utilizing E. coli strain was transformed with plasmids having the altered cysE genes. Then, several transformants overproducing L-cysteine were selected by detecting the halo formation of the L-cysteine auxotroph. The production test of amino acids and analysis of the catalytic property on the mutant enzymes suggest that the carboxy-terminal region of serine acetyltransferase plays an important role in the desensitization to feedback inhibition and the high level production of L cysteine and L-cystine. PMID- 10386616 TI - The human cadherin-10 gene: complete coding sequence, predominant expression in the brain, and mapping on chromosome 5p13-14. AB - In a quest for novel cadherin gene family members in the human dbEST database, an interesting EST clone was identified and chosen for subsequent analysis. Using the technique of 5' rapid amplification of cDNA ends, we isolated the complete coding sequence and a large part of the UTRs of a novel gene. The sequence appeared to correspond to the human cadherin-10 gene, whose sequence was only partially known before. The expression pattern of this cadherin was found to be largely brain-specific, with additional expression in both adult and fetal kidney, and with minor expression in prostate and fetal lung. By FISH analysis the genomic location was determined at human chromosome 5p13-14, which is nearby the reported positions of the human cadherin-6, -12, and cadherin-14 (CDH18) genes. Cadherin-10 shows high relationship to the human cadherin-6 gene. PMID- 10386618 TI - Effect of mutagenesis at serine 653 of Arabidopsis thaliana acetohydroxyacid synthase on the sensitivity to imidazolinone and sulfonylurea herbicides. AB - Resistance to sulfonylurea and imidazolinone herbicides can occur by mutations in acetohydroxyacid synthase (EC 4.1.3.18). Changing serine 653 to asparagine is known to cause insensitivity to imidazolinones but not to sulfonylureas. Here, S 653 of the Arabidopsis thaliana enzyme was mutated to alanine, threonine and phenylalanine. The purified mutated enzymes resemble wild-type in their enzymatic properties. The threonine and phenylalanine mutants are imidazolinone-resistant and the latter is also slightly sulfonylurea-resistant. The alanine mutant remains sensitive to both herbicides. The results suggest that the beta-hydroxyl group is not required for imidazolinone binding and that the size of the side chain determines resistance. PMID- 10386617 TI - Nuclear and nucleolar localization of Saccharomyces cerevisiae ribosomal proteins S22 and S25. AB - Nuclear import usually relies on the presence of nuclear localization sequences (NLSs). NLSs are recognized by NLS receptors (importins), which target their substrates to the nuclear pore. We identified the NLSs of the yeast ribosomal proteins S22 and S25 and studied the former by mutational analysis. Furthermore, in S25 the nucleolar targeting information was found to overlap with its NLS. Comparison with previously published data on yeast ribosomal protein NLSs and computer analysis indicates the existence of a novel type of ribosomal protein specific NLS that differs from the classical Chelsky and bipartite NLSs. The existence of such a ribosomal protein-specific NLS is in accordance with the recent identification of ribosomal protein-specific importins. PMID- 10386619 TI - Alcoholysis reactions from starch with alpha-amylases. AB - The ability of alpha-amylases from different sources to carry out reactions of alcoholysis was studied using methanol as substrate. It was found that while the enzymes from Aspergillus niger and Aspergillus oryzae, two well-studied saccharifying amylases, are capable of alcoholysis reactions, the classical bacterial liquefying alpha-amylases from Bacillus licheniformis and Bacillus stearothermophilus are not. The effect of starch and methanol concentration, temperature and pH on the synthesis of glucosides with alpha-amylase from A. niger was studied. Although methanol may inactivate alpha-amylase, a 90% substrate relative conversion can be obtained in 20% methanol at a high starch concentration (15% w/v) due to a stabilizing effect of starch on the enzyme. As the products of alcoholysis are a series of methyl-oligosaccharides, from methyl glucoside to methyl-hexomaltoside, alcoholysis was indirectly quantified by high performance liquid chromatography analysis of the total methyl-glucoside produced after the addition of glucoamylase to the alpha-amylase reaction products. More alcoholysis was obtained from intact soluble starch than with maltodextrins or pre-hydrolyzed starch. The biotechnological implications of using starch as substrate for the production of alkyl-glucosides is analyzed in the context of these results. PMID- 10386620 TI - A new phagemid vector for positive selection of recombinants based on a conditionally lethal barnase gene. AB - A new phagemid cloning vector for positive selection of recombinants, pBa-7, was constructed which contains an active barnase gene encoding the cytotoxic ribonuclease from Bacillus amyloliquefaciens, under control of the lac promoter. PBa-7 is a derivative of the high-copy number pBluescript II KS+ phagemid in which the modified barnase killer gene has been fused downstream from the lac promoter of the pBluescript II KS+ multiple restriction site. When a lacIq negative Escherichia coli strain is transformed by this vector, the active barnase blocks bacterial growth by massive RNA destruction [1]. However, if barnase is inactivated by insertion of a foreign DNA fragment into the multirestriction site of the vector, this recombinant plasmid no longer interferes with the host viability. The positive selection of recombinant clones is highly efficient and bench manipulations are considerably simplified. When E. coli transformants are plated out on rich medium with ampicillin, only cells containing recombinant plasmids give rise to colonies. In a lacIq-positive host, the positive selection is IPTG-dependent. Therefore, pBa-7 phagemid can be amplified and prepared in large quantities from lacIq-positive E. coli hosts. Hence, pBa-7 seems to be suitable for most genetic engineering manipulations. PMID- 10386621 TI - Arachidonic acid induces the activation of the stress-activated protein kinase, membrane ruffling and H2O2 production via a small GTPase Rac1. AB - Arachidonic acid (AA) is generated via Rac-mediated phospholipase A2 (PLA2) activation in response to growth factors and cytokines and is implicated in cell growth and gene expression. In this study, we show that AA activates the stress activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK) in a time- and dose dependent manner. Indomethacin and nordihydroguaiaretic acid, potent inhibitors of cyclooxygenase and lipoxygenase, respectively, did not exert inhibitory effects on AA-induced SAPK/JNK activation, thereby indicating that AA itself could activate SAPK/JNK. As Rac mediates SAPK/JNK activation in response to a variety of stressful stimuli, we examined whether the activation of SAPK/JNK by AA is mediated by Rac1. We observed that AA-induced SAPK/JNK activation was significantly inhibited in Rat2-Rac1N17 dominant-negative mutant cells. Furthermore, treatment of AA induced membrane ruffling and production of hydrogen peroxide, which could be prevented by Rac1N17. These results suggest that AA acts as an upstream signal molecule of Rac, whose activation leads to SAPK/JNK activation, membrane ruffling and hydrogen peroxide production. PMID- 10386622 TI - Genomic organization of three neurotoxins active on small conductance Ca2+ activated potassium channels from the scorpion Buthus martensi Karsch. AB - According to the known primary sequences of three neurotoxins active on small conductance Ca2+-activated potassium channels from the scorpion Buthus martensi Karsch, their corresponding cDNAs were cloned and sequenced using 3'- and 5' RACE. All of them encoded a signal peptide composed of 28 residues and a mature toxin of 29, 28 and 33 residues, respectively. Their cDNA deduced sequences were totally consistent with those determined, and the C-terminal amidation of one neurotoxin was confirmed. The genomic DNAs of these three toxins were also amplified by PCR, cloned and sequenced. They all consisted of two exons disrupted by a small single intron. All of these introns were inserted within the signal peptide at the same -10 position upstream from the mature toxin, consisting of 94, 78 and 87 bp, respectively. PMID- 10386623 TI - In vivo synthesis of complex N-glycans by expression of human N acetylglucosaminyltransferase I in the filamentous fungus Trichoderma reesei. AB - The human N-acetylglucosaminyltransferase I gene was introduced in the genome of Trichoderma reesei strain VTT-D-80133. Expression was studied after induction from the cellobiohydrolase I promoter. Successful in vivo transfer of GlcNAc was demonstrated by analyzing the neutral N-glycans which were synthesized on cellobiohydrolase I. Final proof of the formation of GlcNAcMan5GlcNAc2 was obtained by NMR analysis. PMID- 10386624 TI - Structure-function analysis of a double-mutant cystic fibrosis transmembrane conductance regulator protein occurring in disorders related to cystic fibrosis. AB - A number of disorders related to cystic fibrosis have been described since the cloning of the cystic fibrosis gene, including infertility due to the congenital bilateral absence of the vas deferens. We have identified, in several patients, complex cystic fibrosis transmembrane conductance regulator genotypes like double mutant alleles. We have now analyzed the structure-function relationships of one of these mutants, R74W-D1270N cystic fibrosis transmembrane conductance regulator, expressed in HeLa cells, to evaluate the contribution of each mutation in the phenotype. We found that R74W cystic fibrosis transmembrane conductance regulator appears to be a polymorphism, while D1270N cystic fibrosis transmembrane conductance regulator could be responsible for the congenital bilateral absence of the vas deferens phenotype. The combination of the two produced a more severe effect on the chloride conductance pathway as well as on the phenotype. PMID- 10386625 TI - Cell-free synthesis of the Ras-binding domain of c-Raf-1: binding studies to fluorescently labelled H-ras. AB - It has previously been shown that the transient kinetics of the interaction between the Ras-binding domain of c-Raf-1 and the proto-oncoprotein Ras can be followed by stopped-flow measurements using the 2',3'-(N-methylanthraniloyl) fluorescence of 2',3'-(N-methylanthraniloyl) guanyl-5'-yl-imidodiphosphate labelled Ras. In continuation of this work, we demonstrate that the His-tagged Ras-binding domain of c-Raf-1 can also be synthesized in a cell-free expression system. After purification by Ni2+ affinity chromatography, His-tagged Ras binding domain of c-Raf-1 could be isolated in sufficient amounts for biochemical and biophysical investigations. The results obtained describe the first example of a cell-free synthesized protein which has been used for stopped-flow measurements to determine the transient kinetics of protein-protein interactions with an effector. PMID- 10386626 TI - Crystal structure of the alpha1beta1 integrin I-domain: insights into integrin I domain function. AB - The alpha1beta1 integrin is a major cell surface receptor for collagen. Ligand binding is mediated, in part, through a 200 amino acid inserted 'I'-domain contained in the extracellular part of the integrin alpha chain. Integrin I domains contain a divalent cation binding (MIDAS) site and require cations to interact with integrin ligands. We have determined the crystal structure of recombinant I-domain from the rat alpha1beta1 integrin at 2.2 A resolution in the absence of divalent cations. The alpha1 I-domain adopts the dinucleotide binding fold that is characteristic of all I-domain structures that have been solved to date and has a structure very similar to that of the closely related alpha2beta1 I-domain which also mediates collagen binding. A unique feature of the alpha1 I domain crystal structure is that the MIDAS site is occupied by an arginine side chain from another I-domain molecule in the crystal, in place of a metal ion. This interaction supports a proposed model for ligand-induced displacement of metal ions. Circular dichroism spectra determined in the presence of Ca2+, Mg2+ and Mn2+ indicate that no changes in the structure of the I-domain occur upon metal ion binding in solution. Metal ion binding induces small changes in UV absorption spectra, indicating a change in the polarity of the MIDAS site environment. PMID- 10386627 TI - Efficient amplification and direct sequencing of mouse variable regions from any immunoglobulin gene family. AB - We have designed two original sets of oligonucleotide primers hybridizing the relatively conserved motifs within the immunoglobulin signal sequences of each of the 15 heavy chain and 18 kappa light chain gene families. Comparison of these 5' primers with the immunoglobulin signal sequences referenced in the Kabat database suggests that these oligonucleotide primers should hybridize with 89.4% of the 428 mouse heavy chain signal sequences and with 91.8% of the 320 kappa light chain signal sequences with no mismatch. Following PCR amplification using the designed primers and direct sequencing of the amplified products, we obtained full-length variable sequences belonging to major (V(H)1, V(H)2, V(H)3, Vkappa1 and Vkappa21) but also small-sized (V(H)9, V(H)14, Vkappa2, Vkappa9A/9B, Vkappa12/13, Vkappa23 and Vkappa33/34) gene families, from nine murine monoclonal antibodies. This strategy could be a powerful tool for antibody sequence assessment whatever the V gene family before humanization of mouse monoclonal antibody or identification of paratope-derived peptides. PMID- 10386628 TI - The negative charge of Glu-127 in protein kinase A and its biorecognition. AB - A set of mutants of protein kinase A (PKA) in which Gln-127 was replaced by Gln, Asp, Asn, and Arg was prepared. Their Km and Vmax values show that the negative charge of Glu-127 (not merely its hydrogen bonding capacity) is indispensable for the kinase activity, since Glu-127/Gln is inactive, in spite of the fact that it can form hydrogen bonds and is very similar in bulkiness and conformation to wt PKA. Glu-127 is involved in the biorecognition of PKA, interacting ionically with the positively charged guanido group of Arg P-3 (a major recognition element in the consensus sequence of PKA). In support of this conclusion, it is shown that a regression of the Glu-127 carboxylate by 1.54 A (as in Glu-127/Asp) results in an active kinase with a similar thermal stability and susceptibility to conformation dependent proteolysis, a similar Vmax, an identical Km for ATP, but a > 20-fold higher Km for kemptide. The two inactive mutants of PKA, Glu-127/Gln and Glu 127/Asn, are potentially useful for studying protein-protein interactions of PKA, e.g. for monitoring enzymatically the displacement of active PKA from its complexes. PMID- 10386629 TI - Characterization of elicitin-like phospholipases isolated from Phytophthora capsici culture filtrate. AB - The phytopathogenic oomycete Phytophthora capsici secretes in culture a phospholipase activity. Two enzyme isoforms exhibiting a high phospholipase B activity were isolated by chromatography and electrophoresis. They differ in their apparent molar masses (22 and 32 kDa). Both proteins are glycosylated and share the same N-terminal amino acid sequence up to the 39th residue with a high homology with capsicein, the P. capsici elicitin. Although devoid of phospholipase activity, capsicein was shown by circular dichroism to specifically interact with negatively charged phospholipids, suggesting that the membrane lipids could be a potential target for elicitins. PMID- 10386630 TI - What is the alpha/beta ratio for prostate cancer? Rationale for hypofractionated high-dose-rate brachytherapy. PMID- 10386631 TI - Influence of the hypoxic subvolume on the survival of patients with head and neck cancer. AB - PURPOSE: Tumor hypoxia is regarded as an important factor influencing radiation response, disease-free, and overall survival of patients with squamous cell carcinoma of the head and neck (SCCHN). This study was performed to reevaluate the prognostic significance of the "classical oxygenation parameters" hypoxic fraction (percentage of pO2 values < 5 mmHg or < 2.5 mmHg, respectively) and median pO2, and to determine the influence of a new radiobiological factor. This factor was termed the "hypoxic subvolume" (HSV) and was defined as percentage of pO2-values below 5 mmHg multiplied by the total tumor volume. The rationale of this parameter was to quantify approximately the amount of hypoxic tissue which should be correlated to the number of hypoxic cells in the tumor. It is obvious that a tumor of 100 cm3 with a hypoxic fraction of 20% (HSV = 20 cm3) contains more hypoxic cells than a tumor of 1 cm3 with a hypoxic fraction of 50% (HSV = 0.5 cm3). METHODS AND MATERIALS: Pretreatment pO2 was assessed in 59 patients with SCCHN with the Eppendorf histograph, and pretreatment volume was determined by ultrasonography (lymphnode metastases) and computer tomography (primaries). All patients were referred to our departments for radiotherapy (n = 27, median dose 70 Gy) or radiochemotherapy (n = 32; 5-FU, mitomycin C, median dose 70 Gy), respectively. All parameters were evaluated using the Kaplan-Meier analysis, and significance was assumed at a p-value of < 0.05 (log-rank test, Cox-Mantel). A multivariate analysis was performed to control for confounding factors. The median follow-up was 233 days. At the time of the evaluation, 34 of the 59 patients were dead. RESULTS: In univariate analyses, the hypoxic fraction (pO2 < 5 mmHg, PO2 < 2.5 mmHg [p < 0.05]), the hemoglobin concentration (p < 0.05), and the hypoxic subvolume (p < 0.01) were of prognostic significance for overall survival. In multivariate analysis, the hemoglobin concentration and the hypoxic subvolume (p = 0.01) were significant prognosticators. We found no significant correlation between tumor volume or median pO2 and overall survival. No clear correlation was found between tumor volume and hypoxic fraction. CONCLUSION: These data suggest that the total amount of hypoxic tissue, as determined by the hypoxic subvolume, influences the prognosis of patients suffering from SCCHN. In addition, our data confirm the statements of previous studies that low pretherapy pO2-values indicate a worse prognosis. PMID- 10386632 TI - Value of computed tomography as outcome predictor of supraglottic squamous cell carcinoma treated by definitive radiation therapy. AB - PURPOSE: To investigate the value of several CT-derived tumor parameters as predictors of local outcome of supraglottic squamous cell carcinoma treated by definitive radiation therapy. METHODS AND MATERIALS: The pretreatment CT studies of 103 patients with supraglottic squamous cell carcinoma were reviewed for tumoral involvement of specific laryngeal anatomic subsites and extralaryngeal tumor spread. After redigitizing the films, tumor volume was calculated with the summation-of-areas technique. Mean follow-up time was 3.4 years. Actuarial statistical analysis of local and locoregional outcome was done for each of the covariates; multivariate analysis was performed using Cox's proportional hazards model. RESULTS: In the actuarial analysis CT-determined primary tumor volume was significantly correlated with local recurrence rate (p < 0.001). Degree of involvement of the paraglottic space at the level of the true vocal cord (p < 0.05) and subglottic extension (p < 0.001) were also significantly correlated with local recurrence rate. In the multivariate analysis, only degree of involvement of the preepiglottic space (p < 0.01) and subglottic extension (p < 0.01) were found to be independent predictors of local recurrence. Total tumor volume was the strongest independent predictor of locoregional failure (p < 0.01). CONCLUSIONS: CT-determined tumor parameters are strong predictors of local and locoregional outcome of supraglottic carcinoma treated by definitive irradiation. PMID- 10386633 TI - Interstitial brachytherapy for stage I and II squamous cell carcinoma of the oral tongue: factors influencing local control and soft tissue complications. AB - PURPOSE: Our aim was to study the treatment parameters that influence local control and soft tissue complications (STC) in a series of 207 Stage I and II squamous cell carcinomas of the oral tongue treated by interstitial brachytherapy (BRT) alone (127 patients), or by a combination using external beam irradiation (EBI) (80 patients) between 1980 and 1993. METHODS AND MATERIALS: The patient distribution was 93 T1, 72 T2a, and 42 T2b. The prescribed BRT dose at the plane 5 mm from the plane of the radioactive sources was 65-70 Gy in BRT alone, and 50 60 Gy in the combined treatment using EBI. Generally, an EBI dose of 30 Gy was used. No prophylactic neck treatment was performed. RESULTS: The 5-year local recurrence-free rate for T1, T2a, and T2b was 92.9%, 81.9%, and 71.8%, respectively (p < 0.05). The lesions of endophytic appearance and those located in the posterior half of the mobile tongue had a significantly lower local control rate than those of other macroscopic appearances (p = 0.02) and those in other localizations (p < 0.01). Most local recurrences (66.7%) occurred within 2 years after treatment. However, 8 of 14 recurrences of T1 and 6 of 15 recurrences among patients treated by BRT alone occurred after 5 years. Statistical analysis showed that, in BRT alone treatment, a dose rate < = 1.0 Gy/h was related to better local control (p = 0.04). There was no significant relationship between BRT dose and local control; however, the incidence of local recurrence was lowest in a BRT dose 65-70 Gy. In the combined treatment, a total dose > 85 Gy (p = 0.01), BRT dose > 55 Gy (p = 0.04), and a dose rate < 0.55 Gy/h (p = 0.03) were significantly related to better local control. The incidence of more severe STC were 11.5% and was significantly higher in T2a (p = 0.03) and T2b (p < 0.01) than in T1. Statistical analysis revealed that a dose rate > = 0.6 Gy/h was significantly related to more STC in BRT alone (p = 0.03), and that a dose rate > = 0.55 Gy/h (p < 0.03) and a BRT dose > 70 Gy ( < 0.05) and a total dose > 100 Gy (p < 0.05) were significantly related to more STC in the combined treatment. Neck metastases occurred in 25% in T1N0, 27% in T2aN0, and 31% in T2bN0 (NS). Eighty eight percent were found within 12 months. Thirty-three secondary cancers including 12 head and neck, 8 esophageal, and 3 gastric were found after treatment. The 5-year crude survival rate for T1, T2a, and T2b was 83.4%, 66.0%, and 70.9%, respectively. CONCLUSION: To acheive better local control and fewer STC, we recommend the following relationships between dose and dose rate. In BRT alone, dose rate should be maintained at < 0.6 Gy/h with a preferable BRT dose 65 70 Gy. In the combined treatment, total dose, BRT dose and dose rate should be kept between > 85 Gy and < = 100Gy, between > 55 Gy and < = 70 Gy, and < 0.55 Gy/h, respectively. We also recommend longer follow-up periods; more than 5 years might be necessary for late local recurrences and for secondary cancers. PMID- 10386634 TI - Carcinoma of the external auditory canal and middle ear. AB - PURPOSE: To evaluate therapeutic modalities used at our institutions regarding local control, disease-free survival and actuarial survival in carcinoma of the external auditory canal and middle ear, in an attempt to provide guidelines for therapy. METHODS AND MATERIALS: A series of 27 patients with carcinoma of the external auditory canal and middle ear treated between 1978 and 1997 in our institutions were analyzed with particular reference to tumor size and its relation to surrounding tissues, patterns of neck node involvement, surgical procedures, and radiation techniques employed. Clinical endpoints were freedom from local failure, overall survival, and disease-free survival. The median follow-up was 2.7 years (range 0.1-17.9 years). RESULTS: Treatment by surgery and radiotherapy resulted in an overall 5-year survival rate of 61%. According to the Pittsburgh classification, the actuarial 5-year survival rate for early disease (T1 and T2 tumors) was 86%, for T3 tumors 50%, and T4 stages 41%. Patients with tumors limited to the external auditory canal had a 5-year survival rate of 100%, patients with tumor invasion of the temporal bone 63%, and patients with tumor infiltration beyond the temporal bone 38%. The rate of freedom from local recurrence was 50% at 5 years. Unresectability by dural and cerebral infiltration, and treatment factors such as complete resection or resection with tumor beyond surgical margins are of prognostic relevance. All patients with dural invasion died within 2.2 years. The actuarial 5-year survival rate of patients with complete tumor resection was 100%, but 66% in patients with tumor beyond surgical margins. 192Iridium high-dose-rate (HDR) afterloading brachytherapy based on three-dimensional computed tomography (3D CT)-treatment planning was an effective tool in management of local recurrences following surgery and a full course of external beam radiotherapy. CONCLUSION: Surgical resection followed by radiotherapy adapted to stage of disease and grade of resection is the preferred treatment of cancer of the external auditory canal and middle ear. PMID- 10386635 TI - American Brachytherapy Society (ABS) recommendations for transperineal permanent brachytherapy of prostate cancer. AB - PURPOSE/OBJECTIVE: To develop and disseminate the American Brachytherapy Society (ABS) recommendations for the clinical quality assurance and guidelines of permanent transperineal prostate brachytherapy with 125I or 103Pd. METHODS AND MATERIALS: The ABS formed a committee of experts in prostate brachytherapy to develop consensus guidelines through a critical analysis of published data supplemented by their clinical experience. The recommendations of the panels were reviewed and approved by the Board of Directors of the ABS. RESULTS: Patients with high probability of organ-confined disease are appropriately treated with brachytherapy alone. Brachytherapy candidates with a significant risk of extraprostatic extension should be treated with supplemental external beam radiation therapy (EBRT). Patient selection guidelines were developed. Dosimetric planning of the implant should be carried out for all patients before seed insertion. A modified peripheral loading is preferred. The AAPM TG-43 recommendations requiring a change in prescription dose for 125I sources should be universally implemented. The recommended prescription doses for monotherapy are 145 Gy for 125I and 115-120 Gy for 103Pd. The corresponding boost doses (after 40-50 Gy EBRT) are 100-110 Gy and 80-90 Gy, respectively. Clinical evidence to guide selection of radionuclide (103Pd or 125I) is lacking. Post implant dosimetry and evaluation must be performed on all patients. It is suggested that the dose that covers 90% (D90) and 100% (D100) of the prostate volume and the percentage of the prostate volume receiving the prescribed dose (V100) be obtained from a dose-volume histogram (DVH) and reported. CONCLUSION: Guidelines for appropriate patient selection, dose reporting, and improved quality of permanent prostate brachytherapy are presented. These broad recommendations are intended to be technical and advisory in nature, but the ultimate responsibility for the medical decisions rests with the treating physician. This is a constantly evolving field, and the recommendations are subject to modifications as new data becomes available. PMID- 10386637 TI - Conventional external-beam radiation therapy alone or with androgen ablation for clinical stage III (T3, NX/N0, M0) adenocarcinoma of the prostate. AB - PURPOSE: To evaluate the outcome of clinical Stage III (T3, N0/NX, M0) prostate cancer treated by conventional radiation alone or with adjuvant androgen ablation. METHODS AND MATERIALS: Three hundred forty-four men with T3, N0/NX, M0 adenocarcinoma of the prostate who received conventional radiation alone (260) or with androgen ablation (84) were analyzed for relapse or rising prostate-specific antigen (PSA), using univariate and multivariate techniques. RESULTS: With a median follow-up of 68 months, the 260 men treated with radiation alone had a 10 year actuarial rate of relapse or rising PSA of 76%. Pretreatment PSA level (< or = 10 ng/ml vs. > 10 < or = 20 ng/ml vs. > 20 ng/ml) and radiation dose (< 68 Gy vs. > or = 68 Gy) were the only independently significant determinants of biochemical failure; Gleason score (2-7 vs. 8-10) was an additional determinant of metastatic relapse. Patients treated to doses < 68 Gy experienced 6-year failure rates exceeding 50% regardless of PSA level. Patients with PSA < or = 10 ng/ml and receiving 68-70 Gy had a 6-year failure of 24%, but those with PSA > 10 ng/ml had relapse rates exceeding 50% even at doses of 70 Gy. At a median follow up of 44 months, the 84 patients treated with radiation and androgen ablation had a 6-year biochemical failure rate of 22%. The only significant determinant of outcome in this group was pretreatment PSA; patients with PSA < or = 80 ng/ml had a 6-year failure rate of only 12% compared to a failure rate of 53% for those with PSA > 80 ng/ml. The outcome for those treated with combined modalities was significantly better than for those treated with radiation alone in all PSA strata. CONCLUSION: Conventional radiation alone has little curative potential for Stage III disease. Doses < 68 Gy are particularly ineffective. Patients with PSA < or = 10 ng/ml may be candidates for conventional radiation to a dose of 70 Gy. Other patients are probably best served by combined radiation-androgen ablation or high-dose conformal radiation. PMID- 10386638 TI - Radiation therapy and patient age in the survival from early-stage breast cancer. AB - PURPOSE: To analyze the use of radiation therapy following local excision of invasive localized breast cancer and subsequent survival by 5-year age category. METHODS: Data for 27, 399 women diagnosed with localized stage of breast cancer and treated with local excision surgery from 1983 through 1992 were collected and provided by the national Surveillance, Epidemiology, and End Results (SEER) program. Use of radiation therapy was analyzed by race, ethnic background, geographic location, and age at diagnosis. Survival for women treated with local excision plus radiation therapy was compared to that of women treated with local excision alone for each 5-year age category. RESULTS: Subjects in older age groups were significantly less likely (p < 0.001) to receive radiation following local excision compared to younger age groups. Statistically significant survival advantages were conferred on women receiving radiation therapy in each 5-year age category from age 35 to 84 years (ranging from p = 0.02 to p < 0.0001). CONCLUSION: While the use of radiation therapy following local excision of early stage breast tumors drops significantly in older age groups, women aged 35-84 years receiving radiation therapy had significant reductions in mortality. These results did not appear to be influenced by the presence of mortal comorbid conditions. These results strongly suggest the need to consider carefully patient characteristics other than age in deciding the course of treatment for early stage breast cancer. PMID- 10386636 TI - Intraoperative optimized inverse planning for prostate brachytherapy: early experience. AB - PURPOSE: To demonstrate the feasibility of an intraoperative inverse planning technique with advanced optimization for prostate seed implantation. METHODS AND MATERIALS: We have implemented a method for optimized inverse planning of prostate seed implantation in the operating room (OR), based on the genetic algorithm (GA) driven Prostate Implant Planning Engine for Radiotherapy (PIPER). An integrated treatment planning system was deployed, which includes real-time ultrasound image acquisition, treatment volume segmentation, GA optimization, real-time decision making and sensitivity analysis, isodose and DVH evaluation, and virtual reality navigation and surgical guidance. Ten consecutive patients previously scheduled for implantation were included in the series. RESULTS: The feasibility of the technique was established by careful monitoring of each step in the OR and comparison with conventional preplanned implants. The median elapsed time for complete image capture, segmentation, GA optimization, and plan evaluation was 4, 10, 2.2, and 2 min, respectively. The dosimetric quality of the OR-based plan was shown to be equivalent to the corresponding preplan. CONCLUSION: An intraoperative optimized inverse planning technique was developed for prostate brachytherapy. The feasibility of the method was demonstrated through an early clinical experience. PMID- 10386639 TI - Quality control in health care: an experiment in radiotherapy planning for breast cancer patients after mastectomy. AB - PURPOSE: The importance of evaluating and improving quality in clinical practice is now generally acknowledged. In this study we estimated different sources of variation in radiotherapy planning for breast cancer patients after mastectomy and sought to test the applicability of a reproducibility and repeatability (R&R) study in a clinical context. METHODS: Eleven radiation oncologists planned radiotherapy three times for three different kinds of breast cancer patients without knowing they were handling the same patient three times. Variation was divided into different components: physicians as operators, patients as parts, and repeated measurements as trials. Variation due to difference across trials (repeatability), that across the physicians (reproducibility), and that across the patients (variability) were estimated, as well as interactions between physicians and patients. Calculation was based on the sum of squares, and analysis was supported by various graphical presentations such as range charts and box plots. RESULTS: Some parts of the planning process were characterized by higher and different kinds of variation than the others. Interphysician variation (i.e., reproducibility) was not high but there were some clearly outlying physicians. The highest variation was in repeatability (= intraphysician variation). The major part of the variation was, however, that from patient to patient: 33% of the total in Parameter 1 and 85% of the total in Parameter 2. CONCLUSIONS: R&R studies are applicable and are needed to evaluate and improve quality in clinical practice. This kind of analysis provides opportunities to establish which kinds of patients require particularly careful attention, which points in the process are most critical for variation, which are the most difficult aspects for each physician and call for more careful description in documents, and which physicians need further training. PMID- 10386640 TI - Rectal cancer and inflammatory bowel disease: natural history and implications for radiation therapy. AB - PURPOSE: There exists little information concerning the natural history of rectal cancer in patients with inflammatory bowel disease (IBD). In addition, the tolerance of pelvic irradiation in these patients is unknown. We analyzed the largest series of patients with IBD and rectal cancer in order to determine the natural history of the disease as well as the effect and tolerance of pelvic irradiation. METHODS AND MATERIALS: A retrospective analysis of 47 patients with IBD and rectal cancer treated over a 34-year period (1960-1994) was performed. Thirty-five patients had ulcerative colitis and 12 patients had Crohn's disease. There were 31 male patients and 16 female patients. The stage (AJC) distribution was as follows: stage 0 in 5 patients, stage I in 13 patients, stage II in 7 patients, stage III in 13 patients, and stage IV in 9 patients. Surgical resection was performed in 44 patients. In two of these patients, preoperative pelvic irradiation was given followed by surgery. Twenty of these patients underwent postoperative adjuvant therapy (12 were treated with chemotherapy and pelvic irradiation and 8 with chemotherapy alone). Three patients were found to have unresectable disease and were treated with chemotherapy alone (2 patients) or chemotherapy and radiation therapy (RT) (1 patient). Radiation complications were graded using the RTOG acute and late effects scoring criteria. Follow-up ranged from 4 to 250 months (median 24 months). RESULTS: The 5-year actuarial results revealed an overall survival (OS) of 42%, a disease-free survival (DFS) of 43%, a pelvic control rate (PC) of 67% and a freedom from distant failure (FFDF) of 47%. DFS decreased with increasing T stage with a 5-year rate of 86% for patients with Tis-T2 disease compared to 10% for patients with T3-T4 disease (p < 0.0001). The presence of lymph node metastases also resulted in a decrease in DFS with a 5-year rate of 67% for patients with NO disease compared to 0% for patients with N1-N3 disease (p < 0.0001). DFS decreased with increasing histopathologic grade with 5-year DFS rates of 71%, 52%, and 24% for grades 1, 2, and 3 respectively (p = 0.03). The T and N stages showed a statistically significant effect on pelvic control, with 5-year PC rates of 60% for Tis-2 versus 26% for T3-4 (p = 0.002) and 79% for NO versus 51% for N1-3 (p = 0.007). The histopathologic grade of the tumor did not significantly affect pelvic control. An analysis of high-risk patients (30) with T3-T4 or N1-N3 disease revealed at 5 years an OS of 9%, a DFS of 10%, a PC rate of 26%, and FFDF of 20%. In this subset of patients, there was a trend toward improved pelvic control in patients receiving RT (14 patients) with a 5-year PC of 60% compared to a rate of 23% for those patients not irradiated (16 patients). Acute complications (grade 3 or >) were noted in three patients (20%) receiving pelvic irradiation +/- chemotherapy and these included two cases of grade 3 skin reactions and one case of grade 4 gastrointestinal toxicity. Two patients (13%) developed small bowel obstruction at 2 and 4 months, respectively, postirradiation which were managed conservatively. There were no long-term complications in patients irradiated. CONCLUSION: Treatment results are comparable to those historically reported for non-IBD-related rectal cancer although the subset of high-risk patients appeared to have a poorer outcome. In light of this finding and the ability of these patients to tolerate chemotherapy and pelvic irradiation, aggressive adjuvant therapy should be given to IBD-associated rectal cancer patients with high-risk features. PMID- 10386641 TI - Conservative management of rectal cancer with local excision and postoperative adjuvant therapy. AB - BACKGROUND: To determine the local control, survival, and functional outcome of local excision plus postoperative therapy for patients with rectal cancer. METHODS: A total of 39 patients underwent a local excision (2 with snare excision of a T1 polyp and 37 with full-thickness local excision) followed by postoperative radiation therapy +/- 5-FU-based chemotherapy. The median follow-up was 41 months, and 11 patients had positive margins. RESULTS: The 5-year actuarial colostomy-free survival was 87% and overall survival was 70%. Crude local failure increased with T stage: 0% T1, 24% T2, and 25% T3. Of the 8 patients (21%) who developed local failure, 5 underwent salvage APR and were locally controlled. Actuarial local failure at 5 years was 31% for T2 disease and 27% for the total patient group. In the 32 patients with an intact sphincter, 94% had good to excellent sphincter function. CONCLUSION: Although local failure in patients with T2 tumors has increased since our prior report, the survival, sphincter function, and local salvage rates are acceptable. Local excision and postoperative therapy remains a reasonable alternative to APR in selected patients. PMID- 10386642 TI - Is prolonged survival possible for patients with supraclavicular node metastases in non-small cell lung cancer treated with chemoradiotherapy?: Analysis of the Radiation Therapy Oncology Group experience. AB - PURPOSE: To determine if patients with non-small cell lung carcinoma (NSCLC) and positive supraclavicular nodes (SN+) have a similar outcome to other patients with Stage IIIB NSCLC (SN-) when treated with modern chemoradiotherapy. METHODS AND MATERIALS: Using the Radiation Therapy Oncology Group (RTOG) database, data were retrospectively analyzed from five RTOG trials studying chemoradiotherapy for NSCLC: 88-04, 88-08 (chemo-RT arm), 90-15, 91-06, 92-04. Comparisons were made between the SN+ and SN- subgroups with respect to overall survival, progression-free survival (PFS), and metastases-free survival (MFS) using the log rank test. Cox multivariate proportional hazards regression analysis was used to determine the effect of several potential confounding variables, including histology (squamous vs. nonsquamous), age (>60 vs. < or = 60), Karnofsky Performance Status (KPS) (<90 vs. > or = 90), weight loss (> or = 5% vs. <5%), and gender. RESULTS: A total of 256 Stage IIIB patients were identified, of whom 47 had supraclavicular nodes (SN+) and 209 did not (SN-). Statistically significantly more SN+ patients had nonsquamous histology (p = 0.05); otherwise, known prognostic factors were well balanced. The median survival for SN+ patients was 16.2 months, vs. 15.6 months for SN- patients. The 4-year actuarial survival rates were 21% and 16% for SN+ and SN- patients respectively (p = 0.44). There was no statistically significant difference in the 4-year PFS rates (19% vs. 14%, p = 0.48). The Cox analysis did not show the presence or absence of supraclavicular nodal disease to be a prognostic factor for survival, MFS, or PFS. The only statistically significant factor on multivariate analysis was gender, with males having a 40% greater risk of mortality than females (p = 0.03). There were no clinically significant differences in toxicity when comparing SN+ vs. SN- patients. Among the 47 SN+ patients, there were no reported cases of brachial plexopathy or other > or = Grade 2 late neurologic toxicity. CONCLUSIONS: When treated with modern chemoradiotherapy, the outcome for patients with supraclavicular metastases appears to be similar to that of other Stage IIIB patients. SN+ patients should continue to be enrolled in trials studying aggressive chemoradiotherapy regimens for locally advanced NSCLC. PMID- 10386643 TI - Radiation therapy morbidity in carcinoma of the uterine cervix: dosimetric and clinical correlation. AB - PURPOSE: To quantitate the impact of total doses of irradiation, dose rate, and ratio of doses to bladder or rectum and point A on sequelae in patients treated with irradiation alone for cervical cancer. METHODS AND MATERIALS: Records were reviewed of 1456 patients (Stages IB-IVA) treated with external-beam irradiation plus two low-dose rate intracavitary insertions to deliver 70 to 90 Gy to point A. Follow-up was obtained in 98% of patients (median, 11 years; minimum, 3 years; maximum, 30 years). The relationships among various dosimetry parameters and Grade 2 or 3 sequelae were analyzed. RESULTS: In Stage IB, the frequency of patients developing Grade 2 morbidity was 9%, and Grade 3 morbidity, 5%; in Stages IIA, IIB, III, and IVA, Grade 2 morbidity was 10% to 12% and Grade 3 was 10%. The most frequent Grade 2 sequelae were cystitis and proctitis (0.7% to 3%). The most common Grade 3 sequelae were vesicovaginal fistula (0.6% to 2% in patients with Stage I-III tumors), rectovaginal fistula (0.8% to 3%), and intestinal obstruction (0.8% to 4%). In the bladder, doses below 80 Gy correlated with less than 3% incidence of morbidity and 5% with higher doses (p = 0.31). In the rectosigmoid, the incidence of significant morbidity was less than 4% with doses below 75 Gy and increased to 9% with higher doses. For the small intestine, the incidence of morbidity was less than 1% with 50 Gy or less, 2% with 50 to 60 Gy, and 5% with higher doses to the lateral pelvic wall (p = 0.04). When the ratio of dose to the bladder or rectum in relation to point A was 0.8 or less, the incidence of rectal morbidity was 2.5% (8 of 320) vs. 7.3% (80 of 1095) with higher ratios (p < or = 0.01); bladder morbidity was 2.3% (7 of 305) and 5.8% (64 of 1110), respectively (p = 0.02). The incidence of Grade 2 and 3 bladder morbidity was 2.9% (10 of 336) when the dose rate was less than 0.80 Gy/h, in contrast to 6.1% (62 of 1010) with higher dose rates (p = 0.07). Rectal morbidity was 2% to 5% in Stage IB, regardless of dose rate to the rectum; in Stages IIA-B and III, morbidity was 5.2% (28 of 539) with a dose rate of 0.80 Gy or less and 10.7% (37 of 347) with higher dose rates (p < 0.01). Multivariate analysis showed that dose to the rectal point was the only factor influencing rectosigmoid sequelae, and dose to the bladder point affected bladder morbidity. CONCLUSIONS: Various dosimetric parameters correlate closely with the incidence of significant morbidity in patients treated with definitive irradiation for carcinoma of the uterine cervix. Careful dosimetry and special attention to related factors will reduce morbidity to the lowest possible level without compromising pelvic tumor control. PMID- 10386644 TI - Long-term follow-up of patients treated with primary radiotherapy for supradiaphragmatic Hodgkin's disease at St. Jude Children's Research Hospital. AB - OBJECTIVE: To assess disease control, patterns of relapse, factors predictive of relapse, and late effects of treatment, we reviewed all cases of supradiaphragmatic (SD) Hodgkin's disease (HD) treated with primary radiation therapy (RT) at our institution. METHODS: We retrospectively reviewed the disease characteristics, treatment history, and long-term outcome of the 106 patients with Stage I and II supradiaphragmatic HD who received definitive irradiation at St. Jude Children's Research Hospital between 1970 and 1995. As of the date of analysis, 95 patients are alive, with a median follow-up of 13.3 years (range, 1.9-24.2 years). RESULTS: The median age at diagnosis was 14.7 years (range, 3.7 22.7). Involved-field RT was given to 13 patients (12%), whereas 37 (35%) had mantle RT, 51 patients (48%) had subtotal nodal irradiation, and 5 (5%) had total nodal irradiation. Relapsed disease developed in 26 patients at a median of 1.8 years (range, 0.2-9.3 years). The 5- and 10-year estimated cumulative incidences of relapse were 20.9% +/- 4.0% and 25.1% +/- 4.3%, respectively. With a median dose of 36 Gy (range, 32-40), in-field failure rate was 6.2%, whereas subdiaphragmatic relapse in sites irradiated prophylactically was 1.5%. There was a trend toward an increased incidence of relapse with higher ESR (p = 0.088) and greater number of sites of disease (p = 0.087). Age, stage, histology, nodal disease > or = 6 cm, the presence of bulky mediastinal disease, and the method of staging did not affect the incidence of relapse. The pattern of failure could not be predicted based on the stage of disease, the extent of subdiaphragmatic staging, the extent of radiation therapy, or the sequence of RT fields-"ping pong" vs. sequential. Subset analysis of Stage II patients revealed significantly more relapses in clinically staged patients. Excluding Stage IA patients with high cervical disease or peripheral nodal disease, nodal extension failures were more common for patients undergoing limited-volume RT, whereas extranodal relapses were likely after STNI or TNI. The estimated 10- and 15-year cumulative incidences of second malignancies were 2.9% +/- 1.6% and 7.9% +/- 3.3%, respectively. Our patients are at increased risk of second malignancies (11 fold), and fatal cardiac (68-fold) and infectious (33-fold) complications. Overall survival at 10 years was 90.8% +/- 3.2%; event-free survival was 72.1% +/ 5.0%. CONCLUSIONS: The current analysis confirms the curative potential of RT for HD in children and adolescents. Despite successful salvage therapy, relapsed disease remained the principal cause of death in our cohort. Excess risk of septic death in asplenic patients, fatal heart disease, and second malignancies may further compromise the ultimate cure of HD in long-term survivors. PMID- 10386645 TI - Postoperative radiotherapy in the management of adult soft tissue sarcoma of the extremities: results with two different total dose, fractionation, and overall treatment time schedules. AB - PURPOSE: This retrospective study was performed to evaluate two postoperative radiotherapy schedules in terms of dose, fractionation, and overall treatment time in soft tissue sarcoma (STS) of the extremities. METHODS AND MATERIALS: Between January 1984 and December 1993, 62 patients with newly diagnosed localized STS of the extremities were treated with maximal conservative surgery and postoperative radiotherapy (RT). Forty-five patients received 50 Gy with conventional fractionation plus a boost dose (5 to 20 Gy). Seventeen patients had hyperfractionated accelerated radiotherapy (HFART) up to a dose of 45 Gy in 3 weeks. RESULTS: With a median follow-up of 72 months, the 5-year local failure rate was 25%, the 5-year disease-free and overall survival rates were respectively 42% and 62%. The 3-year local relapse, disease-free, and overall survival rates were respectively 16%, 44%, and 70% in the conventional radiotherapy group, and 36%, 47%, and 82% in the HFART group (NS). No factor significantly influenced local control with a trend, however, in favor of conventional RT (p = 0.10). CONCLUSION: HFART at the dose of 45 Gy does not seem to be superior to the standard RT schedule, neither in terms of local control, survival, nor in terms of long-term side effects. However this dose could be considered too low as well as the power of comparison between the two groups to draw definitive conclusions. PMID- 10386646 TI - Diode-light transillumination for ophthalmic plaque localization around juxtapapillary choroidal melanomas. AB - PURPOSE: An evaluation of plaque-mounted diode-light transillumination (DLT) for localization of episcleral plaques beneath juxtapapillary tumors. METHODS AND MATERIALS: Two patients scheduled for radiotherapy for juxtapapillary melanomas were offered DLT as an additional method of ophthalmic plaque localization. Plaques were constructed by affixing 4 non-heat producing, light-emitting diodes with their apertures flush with the episcleral outer surface of the plaque's rim. Bio-implantable epoxy was used to encapsulate the electronic components. Then the plaques were loaded with 103Pd seeds. After the eye-plaques were sewn to the episclera covering the base of the intraocular tumors; the diode-lights were illuminated, viewed and recorded. Photodocumentation of the relative position of the 4 lights around tumor's base was obtained in both cases. RESULTS: Digital images of plaque-mounted diode retro-transillumination were obtained. No evidence of diode-light toxicity was noted. Both tumors were found to be covered by the ophthalmic plaques. CONCLUSION: Juxtapapillary tumors are often difficult or impossible to visualize with standard transillumination techniques and have been associated with poor local control rates. We have developed plaque-mounted DLT in an effort to improve ophthalmic plaque localization. Retrobulbar transillumination was viewed by indirect ophthalmoscopy and recorded with video imaging. This technique provides unique photographic documentation of episcleral plaque localization beneath juxtapapillary tumors. PMID- 10386647 TI - Acute central nervous system (CNS) toxicity of total body irradiation (TBI) measured using neuropsychological testing of attention functions. AB - PURPOSE: The purpose of this study was to investigate acute normal tissue damage of low irradiation doses to the healthy, adult central nervous system (CNS) using neuropsychological testing of attention functions. METHODS AND MATERIALS: Neuropsychological testing (IQ, attention [modified Trail-Making Test A, Digit Symbol Test, D2 Test, Wiener Determination Machine]) was used to examine 40 patients (43 +/- 10 years) before and immediately after the first fraction (1.2 Gy) of hyperfractionated total body irradiation (TBI) at the University of Heidelberg. The patients received antiemetic premedication. Test results are given as mean percentiles +/- standard deviation, with 50 +/- 34 being normal. Thirty-eight control patients (53 +/- 15 years) were studied to quantify the influence of hospitalization, stress, and repeated testing. RESULTS: The patients showed normal baseline test results (IQ = 101 +/- 14, attention = 54 +/- 28) and no decrease in test results after 1.2 Gy TBI. Attention functions improved (66 +/ 25) corresponding to a practice effect of repeated testing that was seen in the control group, although alternate versions of the tests were used (IQ = 104 +/- 10, attention before = 42 +/- 29, attention after = 52 +/- 31). CONCLUSION: Our data show no deterioration of neuropsychologic test results acutely after 1.2 Gy whole body exposure in adult patients without CNS disease receiving antiemetic medication. PMID- 10386648 TI - Microvascular and tumor cell alterations during continuous hyperfractionated irradiation: an electron microscopic investigation on the rat R1H rhabdomyosarcoma. AB - PURPOSE: Conventionally fractionated y-irradiation results in severe damage of tumor capillaries associated with decreasing oxygen partial pressure within the tumor. The present study was undertaken to assess whether vasculo-connective changes are less pronounced after continuous hyperfractionated irradiation, implying better tumor oxygenation and improved radiosensitivity. MATERIALS AND METHODS: Twenty rats with an isotransplanted R1H rhabdomyosarcoma were irradiated for 12 days with 2 daily fractions of 2.5 Gy (delta t = 6 h). After 0, 15, 30, 45, and 60 Gy, tumor tissue of 4 rats each was analyzed histologically and electron-microscopically. RESULTS: Untreated rhabdomyosarcomas were composed of spindle-shaped tumor cells with numerous mitoses. There were many apoptotic nuclei and a large central necrosis. Tumor capillaries showed a continuous lining of flattened endothelial cells with broad overlapping cell contacts overlying a delicate continuous basal lamina. During irradiation, mean tumor volume declined from 1.9 cm3 to 1.2 cm3. The number of atypical mitoses and apoptoses increased and numerous giant tumor cells appeared. The proportion occupied by necrotic tumor tissue rose from 30% to 60%. After 15 Gy (3 days), a marked vasodilatation was apparent accompanied by an interstitial edema. Occasionally, endothelial cells were rounded up and showed indented nuclei, with the underlying basal lamina disintegrated. These changes progressed with increasing radiation doses. After 30 Gy (6 days), leukocytes started to adhere to the endothelial wall. Electron-dense fine fibrillar and basal lamina-like deposits appeared in the perivascular space. Endothelial cell edema was only observed after 60 Gy (12 days). Cell contact areas were shortened, however, the endothelial lining was not interrupted. No signs of radiation fibrosis were observed. CONCLUSION: Continuous hyperfractionated irradiation induces relatively discrete alterations of the vasculo-connective tumor tissue as compared to conventional irradiation. This may be an advantage with respect to tumor blood flow, oxygenation, and thus, radiosensitivity. PMID- 10386649 TI - Correlation between gamma-ray-induced G2 arrest and radioresistance in two human cancer cells. AB - PURPOSE: The correlation between radioresistance and gamma-ray-induced G2 arrest was examined in two human cancer cell lines, HeLa (cervical carcinoma) and MeWo (melanoma). METHODS AND MATERIALS: Cellular radioresistance was examined by a colony formation assay and Hoechst 33342 staining. G2 arrest induced by gamma rays was examined by flow cytometry, and the accumulation of cyclin B1 and cdc2 proteins was analyzed using Western blotting. RESULTS: HeLa was more resistant (10% survival dose[D10] = 10 Gy) than MeWo (D10 = 4 Gy) to gamma-rays. In HeLa, cell cycle analysis showed that G2 arrest was induced 10 or 24 h after irradiation of 10 or 4 Gy, respectively. In contrast, no clear G2 arrest in MeWo was observed after irradiation. Western blot analysis showed that cell cycle regulators, cyclin B1 and cdc2, were accumulated in HeLa but not in MeWo. The accumulation of cyclin B1 and cdc2 reached peak levels 24-34 h after irradiation of 10 Gy, and 24 h after irradiation of 4 Gy. In addition, Hoechst staining revealed similar increase in apoptotic bodies with time after irradiation in HeLa and MeWo at isosurvival doses. CONCLUSION: Radioresistance of these human cancer cells is closely correlated with gamma-ray-induced G2 arrest, and cyclin B1 and cdc2 are possible regulators of G2 arrest. PMID- 10386650 TI - The use of active breathing control (ABC) to reduce margin for breathing motion. AB - PURPOSE: For tumors in the thorax and abdomen, reducing the treatment margin for organ motion due to breathing reduces the volume of normal tissues that will be irradiated. A higher dose can be delivered to the target, provided that the risk of marginal misses is not increased. To ensure safe margin reduction, we investigated the feasibility of using active breathing control (ABC) to temporarily immobilize the patient's breathing. Treatment planning and delivery can then be performed at identical ABC conditions with minimal margin for breathing motion. METHODS AND MATERIALS: An ABC apparatus is constructed consisting of 2 pairs of flow monitor and scissor valve, 1 each to control the inspiration and expiration paths to the patient. The patient breathes through a mouth-piece connected to the ABC apparatus. The respiratory signal is processed continuously, using a personal computer that displays the changing lung volume in real-time. After the patient's breathing pattern becomes stable, the operator activates ABC at a preselected phase in the breathing cycle. Both valves are then closed to immobilize breathing motion. Breathing motion of 12 patients were held with ABC to examine their acceptance of the procedure. The feasibility of applying ABC for treatment was tested in 5 patients by acquiring volumetric scans with a spiral computed tomography (CT) scanner during active breath-hold. Two patients had Hodgkin's disease, 2 had metastatic liver cancer, and 1 had lung cancer. Two intrafraction ABC scans were acquired at the same respiratory phase near the end of normal or deep inspiration. An additional ABC scan near the end of normal expiration was acquired for 2 patients. The ABC scans were also repeated 1 week later for a Hodgkin's patient. In 1 liver patient, ABC scans were acquired at 7 different phases of the breathing cycle to facilitate examination of the liver motion associated with ventilation. Contours of the lungs and livers were outlined when applicable. The variation of the organ positions and volumes for the different scans were quantified and compared. RESULTS: The ABC procedure was well tolerated in the 12 patients. When ABC was applied near the end of normal expiration, the minimal duration of active breath-hold was 15 s for 1 patient with lung cancer, and 20 s or more for all other patients. The duration was greater than 40 s for 2 patients with Hodgkin's disease when ABC was applied during deep inspiration. Scan artifacts associated with normal breathing motion were not observed in the ABC scans. The analysis of the small set of intrafraction scan data indicated that with ABC, the liver volumes were reproducible at about 1%, and lung volumes to within 6 %. The excursions of a "center of target" parameter for the livers were less than 1 mm at the same respiratory phase, but were larger than 4 mm at the extremes of the breathing cycle. The inter-fraction scan study indicated that daily setup variation contributed to the uncertainty in assessing the reproducibility of organ immobilization with ABC between treatment fractions. CONCLUSION: The results were encouraging; ABC provides a simple means to minimize breathing motion. When applied for CT scanning and treatment, the ABC procedure requires no more than standard operation of the CT scanner or the medical accelerator. The ABC scans are void of motion artifacts commonly seen on fast spiral CT scans. When acquired at different points in the breathing cycle, these ABC scans show organ motion in three-dimension (3D) that can be used to enhance treatment planning. Reproducibility of organ immobilization with ABC throughout the course of treatment must be quantified before the procedure can be applied to reduce margin for conformal treatment. PMID- 10386651 TI - Static field intensity modulation to treat a dominant intra-prostatic lesion to 90 Gy compared to seven field 3-dimensional radiotherapy. AB - PURPOSE/OBJECTIVE: Recent studies supported by histopathological correlation suggest that the combined use of endorectal magnetic resonance imaging (MRI) and magnetic resonance spectroscopy (MRS) allows differentiation of normal and carcinomatous prostate. The goal of this study was to use static field intensity modulated three-dimensional conformal radiotherapy (SF-IMRT) to treat the entire prostate to a total dose of >70 Gy, while concurrently treating a dominant intraprostatic lesion (DIL) defined by MRI+MRS to 90 Gy while not exceeding normal tissue tolerances. MATERIALS AND METHODS: For the example chosen, the DIL consisted of a large portion of the peripheral zone of the left lobe of the prostate. University of Michigan (UM-PLAN) three-dimensional treatment planning software was used to design a partially shielded 7 field conformal isodose plan that would treat the entire prostate to >70 Gy at 1.8 Gy per day (80% isodose line), while concurrently treating the DIL to 2.25 Gy per day for a total dose of 90 Gy. Dose volume histograms (DVH) were used to compare the rectal doses to rectum and other adjacent normal tissues using these two techniques. RESULTS: SF IMRT as described, allowed a total dose of 90 Gy to encompass the DIL, while the rectal dose was slightly lower than that using the standard 7 field technique to the prostate alone. For example, the dose to 30 cm3 of the rectum was 40 Gy using SF-IMRT and 48 Gy for the standard 7 field technique. Because of differences in the dose per fraction the biologic advantages of the SF-IMRT technique are likely to be even greater. CONCLUSIONS: This study demonstrates the feasibility of using SF-IMRT to treat a DIL involving a single lobe of the prostate, as defined by MRI/MRS, to 90 Gy, while simultaneously treating the prostate to >70 Gy without increasing the dose to surrounding normal tissues. A similar approach could be used to treat multifocal disease. This method of treatment is an alternative to dynamic intensity modulation. It is less expensive, and can be adapted to any radiation therapy department without the use of an inverse treatment planning programs. PMID- 10386652 TI - Variation of clinical target volume definition in three-dimensional conformal radiation therapy for prostate cancer. AB - PURPOSE: Currently, three-dimensional conformal radiation therapy (3D-CRT) planning relies on the interpretation of computed tomography (CT) axial images for defining the clinical target volume (CTV). This study investigates the variation among multiple observers to define the CTV used in 3D-CRT for prostate cancer. METHODS AND MATERIALS: Seven observers independently delineated the CTVs (prostate +/- seminal vesicles [SV]) from the CT simulation data of 10 prostate cancer patients undergoing 3D-CRT. Six patients underwent CT simulation without the use of contrast material and serve as a control group. The other 4 had urethral and bladder opacification with contrast medium. To determine interobserver variation, we evaluated the derived volume, the maximum dimensions, and the isocenter for each examination of CTV. We assessed the reliability in the CTVs among the observers by correlating the variation for each class of measurements. This was estimated by intraclass correlation coefficient (ICC), with 1.00 defining absolute correlation. RESULTS: For the prostate volumes, the ICC was 0.80 (95% confidence interval [CI]: 0.56-0.96). This changed to 0.92 (95% CI: 0.75-0.99) with the use of contrast material. Similarly, the maximal prostatic dimensions were reliable and improved. There was poor agreement in defining the SV. For this structure, the ICC never exceeded 0.28. The reliability of the isocenter was excellent, with the ICC exceeding 0.83 and 0.90 for the prostate +/- SV, respectively. CONCLUSIONS: In 3D-CRT for prostate cancer, there was excellent agreement among multiple observers to define the prostate target volume but poor agreement to define the SV. The use of urethral and bladder contrast improved the reliability of localizing the prostate. For all CTVs, the isocenter was very reliable and should be used to compare the variation in 3D dosimetry among multiple observers. PMID- 10386653 TI - Impact of collimator leaf width on stereotactic radiosurgery and 3D conformal radiotherapy treatment plans. AB - PURPOSE: The authors undertook a study to analyze the impact of collimator leaf width on stereotactic radiosurgery and 3D conformal radiotherapy treatment plans. METHODS AND MATERIALS: Twelve cases involving primary brain tumors, metastases, or arteriovenous malformations that had been planned with BrainLAB's conventional circular collimator-based radiosurgery system were re-planned using a beta version of BrainLAB's treatment planning software that is compatible with MRC Systems' and BrainLAB's micro-multileaf collimators. These collimators have a minimum leaf width of 1.7 mm and 3.0 mm, respectively, at isocenter. The clinical target volumes ranged from 2.7-26.1 cc and the number of static fields ranged from 3-5. In addition, for 4 prostate cancer cases, 2 separate clinical target volumes were planned using MRC Systems' and BrainLAB's micro-multileaf collimators and Varian's multileaf collimator: the smaller clinical target volume consisted of the prostate gland and the larger clinical target volume consisted of the prostate and seminal vesicles. For the prostate cancer cases, treatment plans were generated using either 6 or 7 static fields. A "PITV ratio," which the Radiation Therapy Oncology Group defines as the volume encompassed by the prescription isodose surface divided by the clinical target volume, was used as a measure of the quality of treatment plans (a PITV ratio of 1.0-2.0 is desirable). Bladder and rectal volumes encompassed by the prescription isodose surface, isodose distributions and dose volume histograms were also analyzed for the prostate cancer patients. RESULTS: In 75% of the cases treated with radiosurgery, a PITV ratio between 1.0-2.0 could be achieved using a micro-multileaf collimator with a leaf width of 1.7-3.0 mm at isocenter and 3-5 static fields. When the clinical target volume consisted of the prostate gland, the micro-multileaf collimator with a minimum leaf width of 3.0 mm allowed one to decrease the median volume of bladder and rectum within the prescription isodose surface by 26% and 17%, respectively, compared to the multileaf collimator with a leaf width of 10 mm. Use of the 1.7 mm leaf width micro-multileaf collimator allowed one to decrease the median volume of bladder and rectum within the prescription isodose surface by 48% and 39%, respectively, compared to the multileaf collimator with a leaf width of 10 mm. CONCLUSIONS: For most lesions treated with radiosurgery, the use of a micro-multileaf collimator with a leaf width of 1.7-3.0 mm at isocenter and 3-5 static fields allows one to meet the Radiation Therapy Oncology Group guidelines for treatment planning. Both planning and treatment are relatively straightforward with a micro-multileaf collimator, allowing for efficient treatment of non-spherical targets with either stereotactic radiosurgery or fractionated stereotactic radiotherapy. When the clinical target volume consists of the prostate gland, micro-multileaf collimators with a minimum leaf width of 1.7-3.0 mm allow one to spare more bladder and rectum than one can with a multileaf collimator that has a 10-mm leaf width based on an analysis of PITV ratios, isodose distributions, and dose volume histograms. PMID- 10386654 TI - A reduction in the AAPM TG-36 reported peripheral dose distributions with tertiary multileaf collimation. American Association of Physicists in Medicine Task Group 36. AB - PURPOSE: The American Association of Physicists in Medicine Task Group 36 (AAPM TG-36) data can be used to estimate peripheral dose (PD) distributions outside the primary radiation field. However, the report data apply to linear accelerators not equipped with tertiary multileaf collimators (MLCs). Peripheral dose distributions consist of internal scatter, collimation scatter, transmission through collimation, head leakage, and room scatter. Tertiary MLCs may significantly reduce the PD due to a reduction in collimation scatter, transmission through collimation, and head leakage. Measurements were performed on a multimodality linear accelerator, equipped with a tertiary MLC, to determine PD distributions as a function of energy, field size, distance from the primary radiation field edge, MLC position, and collimator orientation. METHODS AND MATERIALS: Measurements were made using an ionization chamber embedded in a 20 x 40 x 120-cm3 water-equivalent plastic phantom with the secondary collimator and MLC settings of 10 x 10, 15 x 15, 20 x 20, 25 x 25 cm2, and with the MLC fully retracted. Data were taken along the longitudinal axis of the machine for 6 and 18 MV photons. Peripheral dose distributions were evaluated with the collimator set to 180 and 90 degrees. Rotation of the collimator allowed measurements parallel and orthogonal to the direction of motion of the MLC. RESULTS: For both photon energies, peripheral doses measured on a MLC machine were lower than the TG-36 data. When the collimator is rotated by 90 degrees, placing the lower jaws and the MLC leaves along the plane of interest, PD was reduced by as much as a factor of three compared with PDs measured with the MLC fully retracted and the collimator rotated to 180 degrees. PDs measured with the MLC fully retracted and collimator rotated to 180 degrees were comparable to the TG-36 data. Measured PDs were lower when the MLC was used to shape the field than when the MLC was fully retracted. CONCLUSION: A strategic orientation of the collimator with a tertiary MLC can reduce PD distributions by more than a factor of two. This decrease significantly lessens or eliminates the need for external lead shielding to reduce the critical organ dose. This method can be used even when Lipowitz metal blocking (such as for mantle fields) is used, with the MLC leaves oriented along the longitudinal plane. PMID- 10386655 TI - A quality assurance phantom for three-dimensional radiation treatment planning. AB - PURPOSE: Three-dimensional (3D) radiation treatment planning is facilitated through the use of computerized radiation treatment planning systems (RTPSs) and CT simulators (CT-sims). Quality assurance (QA) of these systems is necessary for ensuring that they fulfill their potential. However, comprehensive tools for the systematic QA of these systems have not been developed. We present a phantom that facilitates the evaluation of a large number of nondosimetric functions. These include CT image acquisition and transfer, graphical displays of 3D radiation beams, multiplanar CT image reconstructions, digitally reconstructed radiographs, the representation and manipulation of contoured patient anatomy, dose volume histograms, and the conversion of CT numbers to relative electron densities. METHODS AND MATERIALS: A phantom was constructed which contains materials and geometries that are appropriate for the routine QA of the features described above. The anatomy of the phantom is used as a standard against which the performance of the 3D-RTPS or CT-sim is evaluated. The phantom was used to evaluate three different 3D-RTPSs and a CT-sim at four institutions. RESULTS: Using this phantom, clinically significant errors and limitations in commercially available 3D treatment planning software were discovered. No errors were discovered in the beam display or image reconstructions in the systems examined. Problems were found in the anatomy display, automatic tools, and the CT number to relative electron density conversion data used in some of the systems. CONCLUSION: This phantom is a unique tool designed explicitly for the QA of 3D treatment planning software. Errors and limitations discovered through its use indicate that the QA of commercial treatment planning software is necessary, and that this phantom is an effective device for this task. PMID- 10386656 TI - The optimal radiotherapy schedule for T1 glottic cancers? PMID- 10386657 TI - Anorectal dysfunction following irradiation for uterine carcinoma: comment on Kim et al. IJROBP 41:835-841; 1998. PMID- 10386658 TI - Effects of morphine on metabolism of dopamine and serotonin in brains of alcohol preferring AA and alcohol-avoiding ANA rats. AB - Morphine induces a larger locomotor stimulation in the alcohol-preferring AA rats than in the alcohol-avoiding ANA rats. We have now studied the acute effects of morphine (1 and 3 mg/kg) on metabolism of dopamine and serotonin (5-HT) in the dorsal and ventral striatum of the AA and ANA rats. The basal level of dopamine release, as reflected by the concentration of dopamine metabolite 3 methoxytyramine (3-MT), was lower in the caudate-putamen and nucleus accumbens of the AA rats than in the ANA rats. In the caudate-putamen, morphine increased dopamine metabolism and release more in the AA than in the ANA rats. In the nucleus accumbens and olfactory tubercle, the effects of morphine on dopamine metabolism and release did not differ between the rat lines. Morphine elevated the metabolism of 5-HT in the caudate-putamen and nucleus accumbens of the AA but not in those of the ANA rats. The results suggest that the larger morphine induced psychomotor stimulation of the AA rats in comparison with the ANA rats is associated with the larger effect of morphine on dopamine metabolism in the caudate-putamen and 5-HT metabolism in the caudate-putamen and nucleus accumbens. Furthermore, low basal dopamine release may play a role in the high alcohol preference of AA rats. PMID- 10386659 TI - Influence of alcohol on electrophysiological responses to facial stimuli. AB - A facial discrimination task adapted for use in an event-related potential paradigm was administered to 15 male subjects following oral administration of placebo and 0.56 g/kg alcohol. The stimuli (digital photographs of males and females with happy, sad and neutral facial expressions) generated a series of waves including a prominent positive potential with a latency between 400-550 msec, designated the P450. Three factor repeated measures multivariate analysis of variance was used to evaluate the effect of alcohol on the amplitudes and latencies of the P450 component to the happy and sad faces. As compared to placebo, following alcohol ingestion, male subjects had decreased P450 amplitudes but only to male happy faces compared to female happy faces. These data suggest that this ERP paradigm may be sensitive to detecting subtle effects of alcohol on brain responses to gender-related affective stimuli. PMID- 10386660 TI - Effects of protein kinase C inhibitors on ethanol-induced contractions in isolated rat aorta. AB - The activation of intracellular contractile proteins induces vascular contraction mediated through signal transduction mechanisms. Protein kinase C (PKC) is involved in this signal transduction. The purpose of the present study was designed to investigate the role of PKC on EtOH-, KCl- and phorbol 12, 13 dibutyrate (PDBu)-induced contractions in isolated rat aorta through the use of several different PKC inhibitors. Prior exposure to staurosporine inhibited both EtOH- and KCl-induced contractions in a concentration-dependent manner. The EtOH induced contractions were completely inhibited by staurosporine (5 x 10(-6) M) but complete inhibition of KCl-induced contractions was not observed. Staurosporine (10(-7) M) also significantly inhibited the contractile response to single doses of both EtOH and PDBu. Bisindolylmaleimide (10(-6) M) effectively inhibited contractile responses to both EtOH- and KCl, added cumulatively, and single doses of PDBu. Chelerythrine (10(-7) M) inhibited maximal EtOH-induced contractions. These results suggest that PKC activation plays an important role in the mechanism(s) involved in the contractile activation of rat aorta smooth muscle by EtOH, KCl and PDBu. However, further work is required to elucidate the precise molecular mechanism. PMID- 10386662 TI - [3H]ethylketocyclazocine binding to brain opioid receptor subtypes in alcohol preferring AA and alcohol-avoiding ANA rats. AB - We measured brain regional patterns of [3H]ethylketocyclazocine binding to brain opioid receptors in ethanol-naive alcohol-preferring Alko, Alcohol (AA) and alcohol-avoiding Alko, Non-Alcohol (ANA) rats, by using quantitative autoradiography. This agonist ligand labels all opioid receptor subtypes. The proportions of mu- and delta-opioid receptor binding were evaluated by displacing the mu- and delta-opioid receptor components by the peptides Tyr-D-Ala-Gly N(Me)Phe-Gly-ol (DAMGO, 100 nM) and Tyr-D-Pen-Gly-Phe-D-Pen (DPDPE, 100nM), respectively, the K-component being the naltrexone-sensitive binding left after removal of the above two components. The labeling patterns in the brains of the AA and ANA rats were consistent with the well-known distributions of the opioid receptor subtypes in nonselected rat strains and there was no major difference between the lines. The mu-opioid receptor binding was greater in the AA than ANA rats in several brain regions, most interestingly in the substantia nigra pars reticulata and striatal clusters with elevated shell/core ratios in the nucleus accumbens. The delta-opioid receptor binding did not differ between the lines, whereas the AA rats had more K-opioid receptors than the ANA rats in several brain regions, including limbic areas and basal ganglia. The observed results might indicate altered action of the opioidergic system on dopaminergic pathways in rats with differential alcohol preference. PMID- 10386661 TI - Developmental changes in seizure susceptibility during ethanol withdrawal. AB - It has recently been established that adolescence may represent a developmentally sensitive period with respect to the effects of ethanol, particularly within the NMDA neurotransmitter system. However, the same may also be true of the GABA system. There is evidence to suggest that the number of GABA receptors and their kinetics may change across development. The purpose of this study was to determine if GABA-mediated seizure susceptibility during ethanol withdrawal differed between adolescent and adult animals. Results indicate that adult animals pretreated with ethanol were more likely to achieve maximal tonic-clonic seizure activity and spent more time in stage 3 (or higher) seizure activity than all other groups. These data lend additional support to the contention that adolescent and adult animals differ in their susceptibility to the effects of ethanol and that this susceptibility is transmitter dependent. PMID- 10386663 TI - Prediction of single episodes of drinking during the treatment of alcohol dependent patients. AB - Drinking episodes during the treatment (relapses or lapses) of alcohol-dependent patients is predicted from clinical ratings of patients and individual background data such as alcohol drinking history and social status. The probability of these relapses (or lapses) is determined up to three days in advance using a logistic regression procedure. The study group consisted of 33 male alcohol-dependent persons, who participated in a treatment program. Clinical ratings were performed three times a week by a trained person during a visit to the clinic. The questionnaire contained 23 different items about irritation, craving for alcohol. sleep disturbances, etc. The relapses were either self-reported or detected by a biochemical marker in a urine sample that was taken daily. The most important factor for a relapse in alcohol drinking was shown to be if the patient already had had one relapse during the treatment. Other important clinical factors were the levels of irritation and autonomic disturbances. None of the variables measuring mood shifts was significant. Family conditions during childhood were the most important background variables. The predictions turned out to have a rather high specificity, but the sensitivity was lower. Half of the relapses were not predicted by an increased probability for relapse. Self-reported relapses were predictable from preceding interviews and were also less frequent compared to those detected objectively by the biochemical markers. PMID- 10386665 TI - Infantile handling eliminates reversal learning deficit in rats prenatally exposed to alcohol. AB - Prenatal exposure to ethanol results in learning deficits and alters physiological response to stress. Neonatal handling and stimulation. on the other hand, produce long-lasting physiological and behavioral changes in response to stress. To determine whether early handling, consisting of daily separation and tactile stimulation for the first 3 weeks, can modify fetal alcohol effects on learning ability of young adult rats, offspring of rats chronically exposed to ethanol throughout pregnancy and control animals were trained in a T-maze to learn a position response and then to reverse the learned response. The nonhandled, ethanol-treated rats were deficient on reversal, but the ethanol treated rats that were handled during the first 3 weeks of postnatal development showed no deficit in learning to reverse their previously learned responses. Postnatal handling had no effect on acquisition in alcohol-treated rats. Neither reversal nor acquisition was affected by infantile handling in pair-fed or normal control animals. Early handling may have eliminated the reversal deficit in the ethanol-treated offspring by altering their physiological and behavioral reactivity to stress. PMID- 10386664 TI - Ethanol and lactation: effects of milk lipids and serum constituents. AB - To determine how chronic alcohol administration during lactation affects milk composition and the nutritional status of the dam, EtOH (3 g/kg) as a 20% solution was administered by intubation to Sprague-Dawley rats from days 2 through 15 of lactation. Control dams were pair fed to account for the reduction in food intake observed in the alcohol group, while another control group maintained ad lib food intake. Dams and their litters were weighed daily throughout the study. On day 16, dams were sacrificed and samples taken for further analysis. Blood alcohol levels as well as serum levels of calcium, cholesterol, glucose, iron, lipids, phosphorous, and triglycerides were measured. Liver lipid levels and the total composition and fatty acid profile of the phospholipids in milk were also measured. Results indicate that EtOH administration and pair feeding reduced dam body weight, but not litter growth. Serum iron levels was increased in both EtOH-exposed and pair-fed controls, whereas serum cholesterol was elevated only in EtOH-exposed dams. Finally, of the phospholipids in milk, only one, phosphatidylserine, was slightly but significantly increased by EtOH. If and how these changes impact the development of the offspring remain to be studied. PMID- 10386666 TI - Differential effects of monoaminergic agonists on alcohol intake in rats fed a tryptophan-enhanced diet. AB - The goal of the present study was to determine if enhancement of tryptophan levels in a nutritionally balanced liquid diet would affect alcohol intake in a two-bottle choice procedure. Furthermore. the monoaminergic agonists amphetamine, phentermine (dopaminergic- and noradrenergic-releasing drugs), and fenfluramine (a serotonin releaser) were administered to determine if these drugs reduced alcohol intake in animals fed the tryptophan-enhanced diet compared to those fed an alcohol-containing diet without added tryptophan. Amphetamine 0.5 and 2 mg/kg and phentermine 4 mg/kg selectively reduced alcohol intake in animals fed the tryptophan-enhanced diet; higher doses also reduced alcohol intake in animals fed the control alcohol diet. Three hours after drug administration, phentermine 2 and 4 mg/kg produced increases in consumption of the nonalcoholic diet in animals fed the control diet without affecting consumption in animals fed the tryptophan enhanced diet. Finally, animals in the tryptophan-enhanced group gained less weight than those animals fed an identical diet without the added tryptophan. Neurochemical analysis revealed that the tryptophan-fed groups showed increased 5 HIAA concentrations and serotonin turnover in the striatum. hypothalamus, and frontal cortex compared to animals fed the control diet. The tryptophan-alcohol group also showed almost double the tryptophan levels in the hypothalamus compared to the tryptophan-isocaloric group. These results indicate that, whereas increasing tryptophan levels by itself was not sufficient to alter consumption of an alcohol-containing diet, the administration of monoaminergic agonists significantly interacted with tryptophan in a dose-dependent manner to reduce intake of an alcohol-containing diet without reducing intake of an isocaloric diet. PMID- 10386667 TI - Salvia miltiorrhiza extract inhibits alcohol absorption, preference, and discrimination in sP rats. AB - Experiment 1 of the present study investigated the ability of a standardized extract of Salvia miltiorrhiza in reducing voluntary ethanol intake in ethanol preferring rats of the sP line. Ethanol intake occurred under the two-bottle free choice regimen between 10% (v/v) ethanol and water in daily 4-h scheduled access periods; water was present 24 h/day. Intragastric administration of 200 mg/kg Salvia miltiorrhiza extract resulted in approximately 40% reduction in ethanol intake and preference throughout the 4-day treatment. This effect of Salvia miltiorrhiza extract was likely due to its ability of altering ethanol absorption from the gastrointestinal tract. Indeed, Experiments 2 and 3 of this study demonstrated that 200 mg/kg Salvia miltiorrhiza extract reduced blood ethanol levels (BELs) up to 60% in comparison to control rats, when ethanol was given IG, whereas it failed to modify BELs when ethanol was injected IP. The reducing effect of Salvia miltiorrhiza extract on ethanol absorption may have therefore resulted in an attenuated perception of the psychoactive effects of ethanol sought by ethanol-drinking rats. Consistently, the results of Experiment 4 of the present study demonstrated that a combination of 200 mg/kg Salvia miltiorrhiza extract IG and 1 or 2 g/kg ethanol IG resulted in a partial blockade of the discriminative stimulus effects of ethanol in sP rats trained to discriminate these doses of ethanol from water in a drug discrimination procedure. Collectively, the results are discussed as being suggestive that drugs curbing ethanol absorption from the gastrointestinal tract may constitute a novel strategy for controlling excessive alcohol consumption in human alcoholics. PMID- 10386668 TI - Ethanol intake during lactation impairs milk production in rats and affects growth and metabolism of suckling pups. AB - From parturition, lactating Wistar rats were given 20% alcohol in drinking water and fed a solid diet ad lib (group AL). Pair-fed (PF) and control (C) rats were fed solid diet and given water ad lib (C). All animals were sacrificed on the 12th day of lactation. Ethanol treatment decreased food intake and milk production in lactating rats to a greater level than in PF rats, and a greater reduction in body weight of the AL pups was noted. Brain weight, protein concentration, and DNA content were also lower in pups of AL dams than of PF dams, whereas liver glycogen concentration was higher in the former. Pups from AL dams had higher circulating levels of beta-OH-butyrate, triglyceride, and free fatty acids than those from either C or PF dams. Plasma glucose concentration was lower in both PF and AL than in C pups, whereas the AL group had lower plasma protein concentration than any of the other groups. We conclude that maternal alcohol intake during lactation greatly impairs milk production, and although the known increase of lipid content in milk in rats studied under similar conditions allows an enhanced lipidic components in the pups, this adaptation does not allow normal growth and brain development. PMID- 10386669 TI - Acamprosate decreases the hypermotility during repeated ethanol withdrawal. AB - One of the known behavioral actions of acamprosate is to prevent relapse in weaned alcoholics. However, the mechanism underlying this effect remains unclear. In this study, the motility of Wistar male rats, which were either alcoholized by ethanol inhalation or simultaneously alcoholized and treated orally by acamprosate (400 mg/kg/ day) for 4 weeks, was measured during four episodes of the ethanol withdrawal. The concentrations of excitatory and inhibitory amino acids were also assayed by the microdialysis technique with OPA/BME precolumn derivatisation and electrochemical detection in the nucleus accumbens. Acamprosate reduces both the motility and the glutamate microdialysate content during the first 12 h of ethanol withdrawal in comparison to the alcoholized untreated group. The basal concentration of the sulfonated amino acid taurine increased significantly in alcoholized acamprosate-treated rats compared to alcoholized untreated rats. These results suggest that acamprosate is able to reduce the hypermotility during ethanol withdrawal syndrome directly by reducing the nucleus accumbens glutamate concentration or indirectly by increasing the taurine and GABA brain level. PMID- 10386670 TI - Tetrahydropapaveroline injected in the ventral tegmental area shifts dopamine efflux differentially in the shell and core of nucleus accumbens in high-ethanol preferring (HEP) rats. AB - Since the 1970s tetrahydropapaveroline (THP) and other tetrahydroisoquinoline alkaloids have been implicated in the etiology of alcoholism. When injected into the cerebral ventricle or at specific sites in the mesolimbic system such as the ventral tegmental area (VTA), THP evokes spontaneous and intense intake of alcohol in the nondrinking animal. Further, THP evokes the extracellular efflux of dopamine in the nucleus accumbens (NAC), which comprises, in part, the postulated alcohol drinking "circuit" of neurons. The purpose of this study was to characterize the action of a THP reactive structure, the VTA, on the activity of dopamine and its metabolism in the NAC. In the anesthetized high-ethanol preferring (HEP) rat, artificial CSF was perfused for 10 min at a rate of 10 microl per min specifically in either the core or shell of the NAC. A microbore push-pull cannula system was selected over a microdialysis probe because of its superior recovery of neurotransmitters and tip exposure of less than 1.0 mm. After a series of 5-min perfusions, a single microinjection of 5.0 microg/microl of THP was made in the ipsilateral VTA while the NAC was perfused simultaneously. Sequential samples of the NAC perfusate were assayed by an HPLC coulometric system to quantitate the concentrations of dopamine and its metabolites, DOPAC and HVA, as well as the 5-HT metabolite, 5-HIAA. The results showed that THP injected in the VTA caused a significant increase by 94 +/- 23% in the efflux of dopamine from the core of the NAC. Conversely, the THP injected identically in the VTA suppressed the efflux of dopamine within the shell of the NAC by 51 +/- 10%. The levels of DOPAC, HVA and 5-HIAA within the core and shell of the NAC generally paralleled the increase and decrease in efflux, respectively, of dopamine. CSF control injections in the VTA as well as injections outside of the VTA failed to alter dopamine or metabolite activity in the NAC. These results demonstrate that the presence of THP in the VTA alters directly the function of the pathway of mesolimbic neurons generally and the dopaminergic system specifically. That such a perturbation could account for the induction of alcohol preference is proposed in relation to a reinforcing mechanism involving opioidergic and dopaminergic elements. PMID- 10386671 TI - Emergency department categorization: why SAEM? Why now? Society for Academic Emergency Medicine. PMID- 10386672 TI - Emergency department categorization: solid first steps. PMID- 10386673 TI - Are outpatient admission sources truly a risk factor for appendiceal rupture? PMID- 10386674 TI - In-vivo myocardial substrate alteration during perfused ventricular fibrillation. AB - OBJECTIVES: Earlier work suggests the in-vivo heart alters its substrate utilization as a function of cardiac work. Previous work has also demonstrated the high oxygen requirements of the heart during ventricular fibrillation (VF). The authors hypothesized that myocardial substrate utilization during VF with perfusion is similar to the normal beating heart under conditions of increased workload. METHODS: Myocardial substrate selection was studied in the in-vivo porcine myocardium using 13carbon nuclear magnetic resonance (13C NMR) under conditions of increased cardiac work (dobutamine group) and VF with extracorporeal perfusion (VF group). Once the animal preparation was completed, metabolic steady state was achieved with the infusion of unlabeled acetate into the left anterior descending (LAD) coronary artery. The infused substrate was then changed to [2-13C] acetate and glutamate pool labeling was monitored by 13C NMR. The glutamate C4 resonance areas at baseline and after intervention of either increased workload (dobutamine group) or perfused VF (VF group) were compared within groups using paired t-tests. RESULTS: Baseline aortic and great cardiac vein lactates, glucose levels, blood gases, hemoglobin levels, and temperatures were similar between groups. In both groups, there was a significant decrease from baseline in the labeling of C4 glutamate peaks (dobutamine group: 20.2+/-14.9 vs 84.7+/-32.7, p = 0.002; and VF group: 49.8+/-24.4 vs 83.9+/-24.4, p = 0.02), indicating selection against acetate oxidation in favor of other endogenous substrates. CONCLUSIONS: In the in-vivo heart, despite the absence of functional contractions, changes in substrate utilization during perfused VF are similar to changes that occur with increased workload in the normal beating heart. PMID- 10386675 TI - Alterations in hepatic gluconeogenesis, prostanoid, and intracellular calcium during sepsis. AB - OBJECTIVE: The metabolic alterations observed during sepsis may be associated with changes in local concentrations of intracellular calcium (Ca2+) and prostanoid synthesis in the liver. The authors studied hepatocyte intracellular Ca2+ and the release of glucose and prostanoid in an in-vivo murine liver perfusion model. METHODS: Sepsis was induced in anesthetized, fasted rats by cecal ligation and puncture (CLP, n = 42). Hepatic glucose release was studied in control (n = 10) and CLP (n = 10) groups using a non-recirculating liver perfusion model with and without lactate as gluconeogenic substrate. Hepatocyte intracellular Ca2+ (n = 11) was measured using the selective indicator Fura-2 under basal and epinephrine (10(-5) M) stimulated conditions. 6-Keto prostaglandin F1alpha (6-Keto) and thromboxane B2 (TxB2) were determined from liver perfusate by radioimmunassay (n = 11). Data were analyzed using t-tests and repeated-measures ANOVA. RESULTS: Plasma glucose was significantly lower in CLP groups compared with controls (74.9+/-6.6 vs 115.7+/-4.6 mg/dL, p < 0.05). Plasma lactate was significantly higher in CLP vs controls (3.7+/-0.4 vs 1.4+/-0.1 mM, p < 0.05). Glucose release in isolated perfused livers was significantly lower in CLP vs controls (8.5 vs 16+/-1.2 microM/g/hr, p < 0.001). With the addition of lactate + pyruvate to the perfusate, glucose output in CLP livers was significantly lower following 5 (9.9+/-0.7 vs 17.7+/-1.1 microM/g/hr, p < 0.05) and 10 (11.9+/-1.2 vs 20.6+/-1.3 microM/g/hr, p < 0.001) minutes of perfusion. The basal level of intracellular calcium ([Ca2+]i) in CLP rats (460.1+/-91.6 nM) was significantly higher than in control rats (196.3+/-35.5 nM) (p < 0.05). A significant increase (p < 0.05) in [Ca2+]i occurred after the addition of epinephrine in hepatocytes in control (196.3+/-35.5 vs 331.8+/-41.4 nM) but not CLP (460.1+/-91.6 vs 489.4+/-105 nM) rats. 6-Keto was significantly lower in CLP compared with controls at 30 minutes (25.7+/-3.9 vs 33.4+/-5.5 pg/mL, p < 0.05), whereas TxB2 was not significantly altered (52.1+/-34.7 vs 87.5+/-43.2 pg/mL). CONCLUSION: These results demonstrate that CLP sepsis is associated with an increase in hepatocyte intracellular free Ca2+ concentration along with attenuation of hormone-mediated Ca2+ mobilization and hepatic gluconeogenesis. PMID- 10386676 TI - The risk of appendiceal rupture based on hospital admission source. AB - OBJECTIVE: To determine whether admission source is a potential risk factor for appendiceal rupture. METHODS: Administrative data were obtained from the California Office of Statewide Health Planning and Development for all patients in San Diego County with the primary diagnosis of appendicitis during 1993. The appendiceal rupture ratio was defined as those coded as ruptured (ICD-9-CM codes 540.0 and 540.1) divided by both ruptured and non-ruptured cases (540.9). The odds ratio of appendiceal rupture from routine outpatient office or clinic venues vs those admitted through the ED were calculated using multivariate logistic regression analysis to adjust for age, sex, race, comorbidity, insurance status, and home address to hospital proximity. RESULTS: There were a total of 1,906 patients, of whom 663 (34.8%) had appendiceal ruptures. Of the 1,360 (71.4%) admitted from the ED, 422 (31.0%) had ruptures, compared with 211 (43.3%) of the 487 admitted from outpatient sources (p < 0.0001). Patients with appendicitis directly admitted from outpatient sources were more likely to be complicated by appendiceal rupture than were those admitted through the hospital ED (adjusted odds ratio 1.62, 95% CI = 1.28 to 2.05, p < 0.0001). CONCLUSION: Patients with appendicitis admitted from outpatient sources are more likely to have appendiceal rupture than are those admitted from the ED. PMID- 10386677 TI - The predictive value of endometrial stripe thickness in patients with suspected ectopic pregnancy who have an empty uterus at ultrasonography. AB - Prior research suggests that, in patients with empty uteri at ultrasonography, endometrial stripe thickness may be predictive of ectopic pregnancy or the likelihood of obtaining chorionic villi after a dilatation and evacuation procedure (D+E). However, it is unclear whether the predictive value of endometrial stripe thickness is confined to patients with low beta-human chorionic gonadotropin (beta-hCG) values. OBJECTIVE: To determine whether endometrial stripe thickness is predictive of the risk of ectopic pregnancy or the likelihood of obtaining chorionic villi after D+E in patients with beta-hCG values >1,000 mIU/mL or < or =1,000 mIU/mL. METHODS: In an urban academic ED, the authors conducted a retrospective chart review of consecutive ED patients from August 1991 to August 1997 with abdominal pain or vaginal bleeding, a positive beta-hCG value, and an empty uterus by transvaginal ultrasound examination. Patients were divided into four groups-group 1: endometrium thin, beta-hCG value < or =1,000 mIU/mL; group 2: endometrium thick, beta-hCG value < or =1,000 mIU/mL; group 3: endometrium thin, beta-hCG value >1,000 mIU/ mL; and group 4: endometrium thick, beta-hCG value >1,000 mIU/mL. The secondary analysis was limited to patients who had a D+E performed within 48 hours of the ED visit. The risks of ectopic pregnancy and the likelihoods of obtaining chorionic villi after D+E were compared using chi-square or Fishers' exact test where appropriate, with a p-value of 0.05 being significant. RESULTS: 224 patients were enrolled in the initial analysis. Intergroup differences in the frequency of ectopic pregnancy were of borderline significance (p = 0.08). However, when the comparison was limited to the groups with beta-hCG values < or =1,000 mIU/mL, the predictive value of endometrial stripe thickness reached statistical significance (group 1: 27/99 [27%], group 2: 2/28 [7%], p = 0.05). 79 patients had a D+E performed. Intergroup differences in the rate of obtaining chorionic villi were significant (p = 0.002). Group 1 had the lowest frequency of having chorionic villi identified (4/26 [15%]) and was the only group in which villi were obtained in fewer than 50% of cases. CONCLUSION: Endometrial stripe thickness may be predictive of the risk of ectopic pregnancy and the likelihood of obtaining chorionic villi at D+E. However, its predictive value appears to be confined to patients with beta-hCG values < or =1,000 mIU/mL. PMID- 10386678 TI - Prevalence, therapeutic response, and outcome of ventricular tachycardia in the out-of-hospital setting: a comparison of monomorphic ventricular tachycardia, polymorphic ventricular tachycardia, and torsades de pointes. AB - OBJECTIVE: To investigate out-of-hospital ventricular tachycardia (VT) cardiac arrest patients, comparing the prevalences and outcomes of the following VT subtypes among this population: monomorphic VT (MVT), polymorphic VT (PVT), and torsades de pointes (TdP, PVT with a prolonged QT interval). METHODS: This was a retrospective review from a fire department-based paramedic system of nontraumatic VT cardiac arrest patients (January 1991 to December 1994) with a supraventricular perfusing rhythm (SVPR) at some time during out-of-hospital care, with a measurable QT interval. QT interval was measured from an SVPR, and corrected QT interval (QTc) was calculated and considered prolonged if > 0.45 sec. VT was classified as polymorphic or monomorphic. TdP was defined as PVT with a prolonged QT interval. RESULTS: 196 patients were identified; six were excluded due to incomplete medical records, leaving 190 who met inclusion criteria and were used for data analysis. 117 (62%) patients had MVT, while 73 (38%) patients had PVT; of the 73 patients with PVT, 37 (51%) had normal QTc (non-TdP PVT) and 36 (49%) had prolonged QTc (TdP PVT). 97 (51%) patients had prolonged QTc (PQTc). Regardless of VT type (i.e., MVT vs PVT), 97 (51%) patients had prolonged QTc, with a mean QTc of 0.476+/-0.15 seconds prearrest and 0.464+/-12 seconds postarrest. Patients with PQTc were not more likely to have PVT (70 [37%] vs 76 [40%]; p = 0.705). No significant difference with respect to paramedic-witnessed arrests in each VT morphology group and each QT group was found. The overall hospital discharge rate was 28.4%. Regardless of VT type, patients had similar rates of out-of-hospital return of spontaneous circulation (ROSC) and hospital discharge; patients with PQTc were less likely to be discharged from the hospital (19.6% vs 37.6%; p = 0.01). 27.8% of TdP and 26.8% of non-TdP patients were discharged (p = 0.912). CONCLUSIONS: In this population of out-of-hospital VT arrest patients, MVT is the most common form of VT encountered; PVT and the subtype TdP are also seen in this population with approximately equal frequencies. All three rhythm types demonstrate similar responses to standard Advanced Cardiac Life Support therapy with equal rates of out-of-hospital ROSC and hospital discharge. PQTc may be a marker of poor clinical outcome in patients with out-of-hospital VT arrest. PMID- 10386679 TI - Emergency physician treatment of acute stroke with recombinant tissue plasminogen activator: a retrospective analysis. AB - Stroke teams are advocated for the rapid treatment of patients who have acute ischemic stroke (AIS) with recombinant tissue plasminogen activator (rt-PA). An alternate model uses existing ED resources with specialist consultation as needed. OBJECTIVES: To evaluate the treatment of AIS with rt-PA in this alternate ED model. METHODS: A retrospective observational review was performed of consecutive patients with AIS treated with rt-PA at four hospitals affiliated with an emergency medicine residency. Emergency physicians (EPs) were directly responsible for the treatment of all patients according to predefined guidelines. Records were evaluated from the implementation of the guidelines through December 15, 1997. RESULTS: 37 patients with AIS received rt-PA. Mean age+/-SD was 63+/-16 years (range 22-87), with 25 (68%) male. Patients presented 67+/-29 minutes after stroke onset. After ED arrival, they were seen by the EP in 14+/-13 minutes, had CT in 46+/-22 minutes, and were treated in 97+/-35 minutes. Neurologist consultation occurred in the department for nine patients (24.3%), and by telephone for 14 (37.8%). Symptomatic intracerebral hemorrhage (ICH) occurred in four (10.8%, 95% CI = 0.8% to 20.8%). There were two deaths, neither associated with ICH. Neurologic outcome at discharge compared with presentation in survivors was normal for four patients (11.4%), improved for 16 (45.7%), unchanged for ten (28.6%), and worse for five (14.3%). CONCLUSIONS: In this analysis, EPs, with specialty consultation as required, successfully identified patients with AIS and delivered rt-PA with satisfactory outcomes. Important elements of this model include early patient identification, preestablished protocols, and rapid access to CT scanning and interpretation. PMID- 10386681 TI - A comparison of two automated external defibrillator algorithms. AB - OBJECTIVE: To compare the interval to delivery of the first shock by first responders in mannequin-based cardiac arrest scenarios using two automated external defibrillator (AED) algorithms. METHODS: Thirty-six (18 pairs) of Toronto firefighters (FFs) trained in two AED algorithms, algorithm I (A-I) and algorithm II (A-II), were studied. A-II mandates the immediate application of the AED once pulselessness is established. In contrast to A-I, A-II dictates that no CPR be initiated until it is required by the AED voice prompts. Each FF pair alternated roles while performing "shock-indicated," mannequin-based scenarios according to A-I and A-II. The interval from mannequin contact to delivery of the first shock was recorded. Five pairs were videotaped. The intervals to complete predetermined steps were compared between algorithms to determine in which step(s) time saving occurred. RESULTS: The mean (+/-SD) interval to the first shock in A-I was 80.7 seconds (+/-10.5 sec) (95% CI = 77.2 to 84.2 sec) vs 61.1 seconds (+/-8.75 sec) (95% CI = 58.2 to 64.0 sec) in A-II (p < 0.001). A-II shortened the interval to the first shock by 19.6 sec (+/-11.5) (95% CI = 15.8 to 23.4 sec). The time saving was a direct result of delaying CPR in A-II. CONCLUSION: A-II reduced the interval from mannequin contact to the first shock in standard training scenarios. PMID- 10386680 TI - Adrenal dysfunction in hemodynamically unstable patients in the emergency department. AB - OBJECTIVE: Adrenal failure, a treatable condition, can have catastrophic consequences if unrecognized in critically ill ED patients. The authors' objective was to prospectively study adrenal function in a case series of hemodynamically unstable (high-risk) patients from a large, urban ED over a 12 month period. METHODS: In a prospective manner, critically ill adult patients presenting to the ED were enrolled when presenting with a mean arterial blood pressure < or =60 mm Hg requiring vasopressor therapy for more than one hour after receiving fluid resuscitation (central venous pressure of 12-15 mm Hg or a minimum of 40 mL/kg of crystalloid). Patients were excluded if presenting with hemorrhage, trauma, or AIDS, or if steroids were used within the previous six months. An adrenocorticotropic hormone (ACTH) stimulation test was performed and serum cortisol was measured. Treatment for adrenal insufficiency was not instituted. RESULTS: A total of 57 consecutive patients were studied. Of these, eight (14%) had baseline serum cortisol concentrations of <20 microg/dL (<552 nmol/L), which was considered adrenal insufficiency (AI). Three additional patients (5%) had subnormal 60-minute post-ACTH-stimulation cortisol responses (<30 microg/dL) and a delta cortisol < or =9 microg/dL, which is the difference between the baseline and 60-minute levels. This is functional hypoadrenalism (FH). There were no laboratory abnormalities that distinguished patients with AI or FH from those with preserved adrenal function (PAF). Rates of survival to discharge did not differ between the AI group (7 of 8) and PAF patients (21 of 46; p = 0.052). CONCLUSIONS: Adrenal dysfunction is common in high-risk ED patients. Overall, it has a frequency of 19% among a homogeneous population of hemodynamically unstable vasopressor-dependent patients. The effect of physiologic glucocorticoid replacement in this setting remains to be determined. PMID- 10386682 TI - Ulcers on the fingers. PMID- 10386684 TI - Facilitating the use of the erythrocyte sedimentation rate in the emergency department. PMID- 10386683 TI - Emergency center categorization standards. AB - The SAEM EC Categorization Task Force was developed in response to the 1994 Macy Foundation's recommendation that emergency medicine (EM) organizations "should revise the classification of emergency departments ... to reflect the level of care available in emergency departments, and indicate whether or not facilities are adequate and whether appropriately qualified and credentialed emergency physicians are available 24 hours a day." By holding Level 1 emergency centers (ECs) to objective standards based on the quality of care delivered as well as administrative, research, and educational efforts, SAEM hopes to improve patient care. The SAEM EC Categorization Task Force is now beginning the process of reviewing ECs that provide comprehensive emergency care and serve as regional resources for education, research, and administration in EM. This standards document describes relative and critical criteria to be met in order to receive designation as a Level 1 emergency center. Such centers must meet all critical criteria, and be in sufficient compliance with most or all relative criteria, in order to achieve this designation. This process is entirely voluntary. Any EC is eligible for review. Any institution can initiate the review process by applying. Application materials and further information, including the policies and procedures of the SAEM EC Categorization Task Force, are available from SAEM. PMID- 10386685 TI - Management of a tetraplegic patient in a small rural hospital in Uganda. PMID- 10386686 TI - Case report of an intraperitoneal ruptured abdominal aortic aneurysm diagnosed with bedside ultrasonography. PMID- 10386687 TI - Kidney rupture following extracorporeal shock wave lithotripsy. PMID- 10386688 TI - Reporting clinical prediction rules. PMID- 10386689 TI - NHAMCS: quality of a national emergency department-based information system questioned. National Hospital Ambulatory Medical Care Survey. PMID- 10386691 TI - Male domestic violence victims study: analytical flaws. PMID- 10386690 TI - Intravenous ketorolac vs intravenous prochlorperazine for the treatment of migraine headaches. PMID- 10386692 TI - Rheumatic fever. PMID- 10386693 TI - Complex pulmonary atresia in an adult: natural history, unusual pathology and mode of death. AB - A patient with unrepaired complex pulmonary atresia had a normal life, achieving two successful pregnancies, until the age of 44 years. Confluent central pulmonary arteries were supplied by a fistuious communication from the left coronary artery, and from other collateral arteries arising from the underside of the aortic arch. Unusual aneurysms were present. Death at the age of 46 resulted from dissection and rupture of an aneurysmal dilation of the pulmonary trunk. PMID- 10386694 TI - Experience with the Glenn anastomosis in the adult with cyanotic congenital heart disease. AB - A clinical study on the outcomes of Glenn anastomoses performed since 1987 in eight consecutive patients aged > or = 16 years, and in two performed earlier, showed poor results. One badly selected patient died early as a consequence of high venous pressure, while a further seven had early complications. Seven of eight hospital survivors were followed for 1-10 (median 4.2) years with two deaths (1 and 4 years later). Of the remaining five patients, two improved temporarily, but increased arterial oxygen saturation was not maintained after 6 months. The two patients who had undergone a Glenn anastomosis 10 and 34 years earlier were shown to have pulmonary arteriovenous fistulas. The Glenn anastomosis in these older patients is associated with high rates of complication and appears not to give adequate palliation, particularly when it is the only source of pulmonary blood supply. In the adult, the Glenn anastomosis can be used as a staging procedure for Fontan-type surgery, but must be combined with another source of pulmonary arterial supply. Any adult having a Glenn anastomosis, particularly without another source of pulmonary arterial supply, should be warned of the possibility of worsening of cyanosis and symptoms. The second stage of the procedure may need to be performed soon after the first should the hypoxia prove intolerable. PMID- 10386695 TI - Acute complications in the current era of therapeutic cardiac catheterization for congenital heart disease. AB - The acute complications of therapeutic cardiac catheterization for congenital heart disease as performed currently in a small unit were reviewed. In recent years, there has been a significant increase in the number of lesions thought amenable to catheter therapy. Only a few reports, however, have addressed the overall incidence of acute complications of therapeutic cardiac catheterization, all representing the experience of centres performing moderate-to-large numbers of procedures. A retrospective review was performed of 425 therapeutic catheter procedures performed at our institution between May 1993 and November 1997. Acute complications were retrieved from the database. This included all adverse events that were clinically recognized at the time of or within 2 weeks after the procedure and which, to the best of the authors' clinical judgement, were related to the catheterization and not part of the natural history of the child's illness. All patients were observed overnight following the procedure, and stayed in hospital if a complication developed. There were 49 acute complications (11.5%), of which 43 (10.1%) were deemed minor and 6 (1.4%) were considered major. The rate was low in patients with valvar pulmonary stenosis, including three neonates (3/45, 6.7%), for those undergoing angioplasty of native co arctation (1/15, 6.7%) and pulmonary arteries (2/27, 7.4%); and for coil embolization of systemic to pulmonary collateral arteries (1/16, 6.3%). The rate was high in patients with valvar aortic stenosis, including 12 neonates (9/37, 24.3%), and for angioplasty of re-coarctation (4/23, 21.7%). There were more overall complications in neonates (25.6%) than in older patients (10.1%) (p < 0.01). Two patients died (0.5%), but no patient required emergency surgical intervention. In spite of the introduction of many new therapeutic modalities with greater intrinsic risk, and the fact that patients with more complex lesions and who are more acutely ill are being treated, the overall rate of complications remains relatively low. This probably reflects improvements in pericatheterization medical management, in selection of patients, in procedural techniques, and in the experience of operators. PMID- 10386696 TI - Tachycardias in children originating in the right ventricular outflow tract: lack of clinical features predicting the presence and severity of the histopathological substrate. AB - The aim was to determine whether the clinical features of tachycardias originating from the right ventricular outflow tract in children with an apparently normal heart could predict the presence and the severity of the histopathological substrate. Thirteen children (median age 6 years; range 6 months-12 years) with tachycardia originating from the right ventricular outflow tract of apparently normal hearts, were assessed by echocardiography, heart catheterization with angiography, endomyocardial biopsy (13 patients) and magnetic resonance imaging (MRI) (nine patients). Tachycardia was symptomatic in six and sustained in nine. Endomyocardial biopsy and MRI revealed acute myocarditis in five patients (38%), fatty infiltration of the right ventricle in two (15%), and minor histologic abnormalities in three (23%). Myocarditis was diagnosed in three of nine patients with sustained ventricular tachycardia, as opposed to two of four with non-sustained tachycardia (p = NS); in three of six symptomatic versus two of seven asymptomatic patients (p = NS); and in two of eight patients in whom ventricular tachycardia was induced during exercise testing as opposed to one of three in which it was not inducible (p = NS). A histopathological substrate was found in six of nine patients with sustained ventricular tachycardia, and in all four with non-sustained tachycardia (p = NS); in five of six patients with symptoms versus five of seven asymptomatic patients (p = NS); and in five of eight with inducible ventricular tachycardia during exercise testing versus all three in whom it was not inducible (p = NS). The mean rate of tachycardia was 184+/-39 beats min(-1) in patients with myocarditis, as opposed to 171+/-48 in patients without myocarditis (p = NS); and 163+/-33 in patients with a histopathological substrate compared with 210+/-65 in patients without a histopathological substrate (p = NS). It is concluded that a histopathological substrate is present in the greater majority of children affected by the so-called right ventricular outflow tract tachycardia, but that the clinical features of the tachycardia do not predict the presence and the severity of this histopathological substrate. PMID- 10386697 TI - Sternal wound and mediastinal infections in infants with congenital heart disease. AB - The objective was to describe the epidemiologic, clinical, bacteriologic and therapeutic features of seven infants who developed sternal wound and mediastinal infections following palliation and/or repair procedures for congenital heart disease. A retrospective chart review was used. All infants with sternal wound and mediastinal infections were < 30 days of age at the initial operative procedure. Six of the infants had hypoplastic left heart syndrome, and one had complete transposition. Two infants required delayed closure of their chest wound. Three infants had superficial sternal infections and presented at a mean of 12 days postoperatively. Four infants had infection of the deep mediastinal structures: they were all asymptomatic and had purulent collections in their mediastinum at their second palliative operation, which was performed at a mean of 120 days after the initial surgery. Staphylococcus aureus, or coagulase negative Staphylococcus, was isolated from the wound and/or blood of six infants. All infants with mediastinal infections were managed with operative debridement. Infants with superficial infections underwent local debridement. All infants received long-term intravenous antibiotics. Mediastinal infections in infants undergoing palliative staged procedures for congenital heart lesions may be chronic and indolent, resulting in delayed repair of congenital heart lesions. PMID- 10386698 TI - Persistence of the left superior caval vein: can it potentiate obstructive lesions of the left ventricle? AB - Recent evidence has suggested that persistence of the left superior caval vein is associated with a high incidence of obstructive lesions of the left heart. To shed more light on this issue 1085 patients with congenital heart disease were studied retrospectively, with the aim of estimating the prevalence of a persistent left superior caval vein and its associated anomalies, focusing attention on obstructive lesions in the left and right ventricles. Patients with isomerism of the atrial appendages, or hypoplastic left heart syndrome, were excluded. A persisting left superior caval vein was present in 57 patients (5.2%). The overall incidence of obstructive lesions of the left heart was higher in patients with than in those without a persistent left superior caval vein (31.6 versus 7.8%,p < 0.001). Relative hypoplasia of the left ventricle was also higher in patients with persistent left superior caval vein (14 versus 0.8%, p < 0.001). The obstructive lesions found in the left heart, compared with the number in those without a left caval vein, were: mitral stenosis, 5.2 versus 0.7%; subaortic stenosis, 5.3 versus 0.9%; aortic coarctation, 17.5 versus 5.8% (p < 0.01); all of these in association, 3.5 versus 0.4%. In contrast, the incidence of obstructive lesions of the right heart was similar in the two groups of patients. It is concluded that persistence of the left superior caval vein can perturb the normal development of the left ventricle, being strongly associated with obstructions to left ventricular inflow and outflow. PMID- 10386699 TI - The Italian Multicentric Study on Epidemiology of Congenital Heart Disease: first step of the analysis. Working Party of the Italian Society of Pediatric Cardiology. AB - We present the aims, methodology and initial results from the Italian Multicentric Study for the registration and follow-up of congenital heart disease. The general aims are to measure the prevalence of congenital heart disease in different geographic areas of Italy, and to assess the survival and outcome of affected babies. During the years 1992 and 1993, eighteen centers for Pediatric Cardiology spread all over the Country enrolled 1445 new babies with congenital cardiac malformations from a population of 341,647 surveyed livebirths. The new cases were registered using the same methodologic criteria of the EUROCAT study in order to evaluate differences and/or similarities between the studies. The prevalence varied between 1.8% and 8.1%; the average being 4.6%. The large range in prevalence is presumed to be related to different customs and hierarchies in flow and referral of patients. We provide total prevalence of individual lesions, and distribution of sentinel cardiac anomalies, in the Italian study and compare them with EUROCAT. Isolated ventricular septal defect is the most common lesion (39%); followed by atrial septal defect (7.5%); pulmonary valvar stenosis (7.3%); atrioventricular septal defects (5.4%); patency of the arterial duct (3.8%); complete transposition (3.7%); tetralogy of Fallot (3.3%); aortic coarctation (2.4%); aortic valvar stenosis (2.2%); and left heart hypoplasia (1.8%). The echographic stratification of ventricular and atrial septal defects, by location and size, was in keeping with the findings of the EUROCAT study. Because of the recent widespread availability of color-Doppler techniques, the stratification of aortic and pulmonary valvar stenosis was an innovative approach in our study. Among the complex cardiovascular anomalies, double inlet ventricle and pulmonary atresia had a proportion of about 2% each; with double outlet right ventricle, common arterial trunk, Ebstein's malformation, tricuspid atresia, interrupted aortic arch and totally anomalous pulmonary venous connection having a proportion ranging from 0.5 to 0.8%. We discuss clinical features, such as frequency of extracardiac anomalies and familial aggregation of congenital heart disease, in comparison with the EUROCAT data. PMID- 10386700 TI - Asymmetrically short tendinous cords causing congenital tricuspid regurgitation: improved understanding of tricuspid valvar dysplasia in the era of color flow echocardiography. AB - BACKGROUND: Tricuspid regurgitation as a manifestation of an isolated congenital anomaly of the tricuspid valve is rare. Cross-sectional and color Doppler echocardiography allow improved evaluation of tricuspid valvar function. As a result, the heterogeneous category of congenital tricuspid valvar dysplasia may be better understood from a functional point of view. We are reporting a distinct entity in which tricuspid valvar regurgitation results from failure of coaptation due to short tendinous cords tethering the septal leaflet. PATIENTS AND RESULTS: Three children with significant primary tricuspid regurgitation were evaluated, treated, and followed. On echocardiographic evaluation, a central regurgitant jet of moderate or severe degree was directed toward the atrial septum through poorly coapting tricuspid valvar leaflets, which did not approximate due to tethering of the septal leaflet by abnormally short cords. In one patient, the tricuspid valve was otherwise normal; in the other two the leaflets and cords were also thickened. Two patients underwent surgery at 9 and 11 years of age. The cords tethering the septal leaflet were augmented by interposing appropriate lengths of expanded polytetrafluoroethylene suture and performing commissural annuloplasty. Both patients are asymptomatic 33 and 42 months postoperatively, with mild residual tricuspid regurgitation that has not changed since surgery. The other patient, an 8 month-old infant, has not yet undergone surgery. CONCLUSIONS: Asymmetric tendinous cords of the tricuspid valve causing tethering of the septal leaflet is a distinct cause of tricuspid regurgitation that can be recognized with echocardiography. Although rare, the importance of recognizing this lesion lies in its being readily amenable to surgical repair. PMID- 10386701 TI - Echocardiographic diagnosis of totally anomalous pulmonary venous connection to the azygos vein. AB - Totally anomalous pulmonary venous connection to the azygos vein is a rare congenital heart malformation in which all the pulmonary venous blood returns anomalously to the azygos vein. Among 111 consecutive patients with totally anomalous pulmonary venous connection undergoing surgical correction at our institution between June 1982 and September 1997, this malformation was present in seven cases. By echocardiography, using a subxyphoid short-axis view at the atrial level and a modified suprasternal sagittal view, the malformation was diagnosed when the pulmonary venous confluence was traced posteriorly and superiorly relative to the right pulmonary artery and right bronchus, finally reaching reach the superior caval vein. Totally anomalous pulmonary venous connection to the azygos vein was misdiagnosed in the first two patients, both by echocardiography and angiocardiography. In the subsequent five patients, a precise diagnosis was obtained by echocardiography. Echocardiography, therefore, can be considered an accurate diagnostic tool permitting recognition of totally anomalous pulmonary venous connection to the azygos vein, and permitting corrective surgery without recourse to catheterization and angiography. PMID- 10386702 TI - Isolated connection of the right superior caval vein to the left atrium: non invasive neonatal diagnosis. AB - Isolated anomalous drainage of the right superior caval vein to the left atrium is a very rare cause of cyanosis in the newborn. Herein, the cross-sectional echocardiography and colour Doppler findings of this malformation are described. PMID- 10386703 TI - Palliative arterial switch for complete transposition with ventricular septal defect. AB - A 28-year-old female patient with complete transposition, ventricular septal defect and persistence of the arterial duct underwent a palliative arterial switch procedure in 1976 at 7 years of age. Therefore, she has survived for 22 years and lives a near normal life. She is married, has been counselled against pregnancy and has increasing cyanosis with the typical features of the Eisenmenger syndrome. PMID- 10386704 TI - Double-switch operation for congenitally corrected transposition and Ebstein's malformation. AB - An infant is described with congenitally corrected transposition and Ebstein's malformation. Banding of the pulmonary trunk had been previously performed because of a muscular ventricular septal defect. The patient underwent the double switch procedure with the intention of unloading the morphologically right ventricle and the malformed tricuspid valve. This resulted in prompt postoperative functional and haemodynamic improvement. PMID- 10386705 TI - An unusual type of combined aortic coarctation due to fibromuscular dysplasia. AB - A 3-year-old patient is described with an unusual form of co-arctation due to hypoplasia of the transverse arch and fibromuscular dysplasia involving a long segment of the thoracic aorta. Surgical repair required resection of the aorta from the distal transverse arch to the mid-descending thoracic aorta, and replacement with a 16-mm Dacron interposition graft. This case demonstrates the importance of preoperative evaluation of the entire aorta in the presence of co arctation due to fibromuscular dysplasia. PMID- 10386706 TI - Lobar emphysema due to anomalous aortic origin of the left pulmonary artery. AB - The unusual case of an infant with aortic origin of the left pulmonary artery is presented. The patient developed a rare complication of lobar emphysema due to bronchial compression from the enlarged right pulmonary artery. Operative anastomosis of the left pulmonary artery to the pulmonary trunk was successful, with subsequent resolution of the lobar emphysema. PMID- 10386707 TI - Hypoplastic left heart syndrome with right aortic arch, bilateral arterial ducts and origin of the left subclavian artery from the left pulmonary artery. AB - The rare association, in a left-sided heart with hypoplastic left heart syndrome, of right aortic arch, bilateral patent arterial ducts and origin of the left subclavian artery from the left pulmonary artery are described. Cardiac catheterization was performed because of the abnormal anatomy of the arch noted at echocardiographic examination. This abnormality is of surgical importance when planning the Norwood operation. PMID- 10386708 TI - Ascites and weight loss in a child: due to congenital division of the right atrium. AB - Congenital division of an atrial chamber is a very rare congenital malformation that more commonly affects the left atrium but which may, in rare circumstances, involve the right atrium. Such a divided right atrium may present with symptoms consistent with increased portal venous pressure. Reported is a case with unusual clinical presentation. The patient underwent resection of the dividing shelf with good postoperative results. PMID- 10386709 TI - Absence of aortic valvar leaflets with normally related great arteries and stiff left ventricle. PMID- 10386710 TI - Sentinel node biopsy as a surgical staging method for solid cancers. AB - The sentinel node is the first lymph node that drains a primary tumour. If this lymphatic drainage occurs in a step-wise fashion, this lymph node reflects the pathological status of the remaining lymph node basin. The day before the operation, a total dose of 60 MBq 99mTc nanocolloid is injected around the primary tumour for lymphoscintigraphy. On the day of surgery, 1 ml of blue dye is injected around the primary tumour to facilitate sentinel lymph node detection. After making a small incision over the regional lymph node region, the sentinel node can be detected using a hand-held gamma ray detection probe; the sentinel lymph node and the afferent lymphatic vessels will be stained blue. Sentinel node biopsy has proved useful for malignant melanoma, breast cancer, penile cancer, vulvar cancer, Merkel cell carcinoma and thyroid cancer. New studies are described on breast cancer and malignant melanoma. Gamma-probe-guided localization of radiolabelled lymph nodes can direct the surgeon non-invasively to the exact location of the sentinel node. Once localized with a gamma probe, it is quick and easy to remove the sentinel node through a small incision. Discriminating the node from other tissue can be aided by blue dye which stains the lymph node. It appears that both radioactivity and blue dye are complementary for locating the sentinel node. PMID- 10386711 TI - How can laparoscopic management assist radiation treatment in cervix carcinoma? AB - PURPOSE: To determine the role of laparoscopic lymphadenectomy (pelvis +/- para aortic nodes) and laparoscopic hysterectomy in cervical cancer compared to 'classic radical surgery' in patients undergoing surgery in comparison with modern imaging in patients treated with radiotherapy alone. MATERIALS AND METHODS: The limitations of modern imaging are presented as well as how complication rates can be increased when classic laparotomy is followed by radiation therapy. Laparoscopic procedures are described with particular emphasis on how to provide information on lymph node metastases with the risk of overlooking microscopic involvement. A number of clinical experiences are cited to illustrate this problem and show how treatment approaches can be adapted. RESULTS: The role of laparoscopy is evaluated according to different clinical situations and treatment protocols emphasizing the possibilities offered by this method to the radiotherapist. CONCLUSION: When developing laparoscopic techniques for the management of cervical carcinoma, caution must be exercised to ensure that these techniques are not detrimental to the prognosis. PMID- 10386712 TI - Long-term results of preoperative intra-arterial doxorubicin combined with neoadjuvant radiotherapy, followed by extensive surgical resection for locally advanced soft tissue sarcomas of the extremities. AB - BACKGROUND AND PURPOSE: In the 1980s a combined modality therapy of intraarterial doxorubicin, neoadjuvant radiotherapy and surgery was initiated at the Groningen University Hospital as a limb-saving treatment for locally advanced, primarily irresectable high-grade soft tissue sarcomas (STS) of the extremities. This study presents the short- and long-term results. PATIENTS AND METHODS: Between 1983 and 1987, 11 patients were treated with intraarterial doxorubicin, preoperative radiotherapy (10 x 3.5 Gy) and surgical resection. Non-radical resections received additional postoperative radiotherapy of 20-30 Gy. RESULTS: The limb salvage rate was 91%, without local recurrences during a median follow-up of 84 months. Six patients died (55%); five from metastatic disease (45%). There were five long-term survivors with a median follow-up of 10 years. Three patients (60%) suffered serious late complications, resulting in disabilitating limb function. CONCLUSION: Although this approach is feasible as a limb-saving treatment for these unfavorable STS, long-term morbidity is high. PMID- 10386713 TI - Late course accelerated fractionation in radiotherapy of esophageal carcinoma. AB - PURPOSE: To evaluate the efficacy of adding accelerated fractionation after completing two thirds of routine fractionated radiotherapy in esophageal carcinoma. METHODS AND MATERIALS: From April 1988 to April 1990, 85 patients with histologically confirmed carcinoma of the esophagus were randomized into two groups. (1) The conventional fractionation (CF) group, received 1.8 Gy per day five times a week to a total dose of 68.4 Gy in 7-8 weeks, and (2) the late course accelerated hyperfractionated (LCAF) group which received the same schedule as the CF group during the first two thirds of the course of radiotherapy to a dose of 41.4 Gy/23 fx/4 to 5 weeks. This was then followed by accelerated hyperfractionation using reduced fields. In the LCAF portion of the radiotherapeutic course, the irradiation schedule was changed to 1.5 Gy twice a day, with an interval of 4 h between fractions, to a dose of 27 Gy/18 fx. Thus the total dose was also 68.4 Gy, the same as the CF group, but the course of radiotherapy was shorter, being only 6.4 weeks. The same Cobalt 60 teletherapy unit was used to treat all the cases. RESULTS: The 5 year actuarial survival and disease-free survival rates in the LCAF group were 34% and 42%, as compared to 15% and 15% respectively in the CF group, all statistically significant. Better local control was seen in the LCAF group than in the CF group, the 5 year control rates being 55% versus 21% (P = 0.003). The acute reactions were increased but acceptable in the LCAF patients, the radiation treatments could be completed without any breaks. The late reactions as observed after 5 years were not increased in comparison with the CF patients. CONCLUSIONS: The results from this study show that the late course accelerated hyperfractionated radiotherapy regime can improve results in esophageal carcinoma, with acceptable acute reactions as compared to conventional radiotherapy. PMID- 10386714 TI - Pre-irradiation carboplatin and etoposide and accelerated hyperfractionated radiation therapy in patients with high-grade astrocytomas: a phase II study. AB - PURPOSE: To investigate feasibility, activity and toxicity of pre-irradiation chemotherapy (CHT) in patients with newly diagnosed high-grade astrocytoma. MATERIAL AND METHODS: Thirty-five patients with glioblastoma multiform (GBM) and ten patients with anaplastic astrocytoma (AA) entered into this study. Three weeks after surgery patients started their CHT consisting of two cycles of carboplatin (CBDCA) (C) 400 mg/m2, day 1 and etoposide (VP 16) (E) 120 mg/m2, days 1-3, given in a 3-week interval. One week after the second cycle of CE, accelerated hyperfractionated radiation therapy (ACC HFX RT) was introduced with tumor dose of 60 Gy in 40 fractions in 20 treatment days in 4 weeks, 1.5 Gy b.i.d. fractionation. RESULTS: Responses to two cycles of CE could be evaluated in 29 (67%) of 43 patients who received it. Fourteen patients were found impossible to determine radiographic response due to an absence of post-operative contrast enhancement because they were all grossly totally resected. There were 7, 24% (95% confidence intervals - CI, 9-40%), PR (2 AA and 5 GBM), 19 SD, and 3 PD. After RT, of those 29 patients, there were 3 CR and 11 PR (overall objective response rate was 48% (95% CI, 30-67%)), 12 SD, and 3 PD. Median survival time (MST) for all 45 patients is 14 months (95% CI, 11-20 months, while median time to progression (MTP) for all patients is 12 months (95% CI, 8-16 months). Toxicities of this combined modality approach were mild to moderate, with the incidences of CHT-induced grade 3 leukopenia, being 5% (95% CI, 0-11%), and grade 3 thrombocytopenia being 7% (95% CI, 0-15%). Of RT-induced toxicity, grade 1 external otitis was observed in 26% (95% CI, 13-39%), while nausea, vomiting and somnolence were each observed in 5% (95% CI, 0-11%) patients. CONCLUSION: Pre irradiation CE and ACC HFX RT was a feasible treatment regimen with mild to moderate toxicity, but failed to improve results over what usually would be obtained with 'standard' approach in this patient population. PMID- 10386715 TI - Ischemic heart disease after mantlefield irradiation for Hodgkin's disease in long-term follow-up. AB - BACKGROUND AND PURPOSE: In patients with Hodgkin's disease treated by radiotherapy with a moderate total dose and a low (mean) fraction dose to the heart, the risk of ischemic heart disease was investigated during long-term follow-up. MATERIALS AND METHODS: The medical records of 258 patients treated in the period 1965-1980 with radiotherapy alone as the primary treatment were reviewed. The median follow-up was 14.2 years (range 0.7-26.2). The mean total dose and fraction dose to the heart were 37.2 Gy (SD 2.9) and 1.64 Gy (SD 0.09), respectively. The impact on the development of ischemic heart disease of treatment-related parameters, such as the applied (fraction) dose, irradiation technique (one or two fields per day), and chemotherapy in case of a relapse, was investigated. The incidence of ischemic heart disease in this patient population was compared with the expected incidence based on gender, age and calendar period specific data for the Dutch population. RESULTS: Thirty-one patients (12%) experienced ischemic heart disease (actuarial risk at 20-25 years: 21.2% (95% C.I. 15-30). Twenty-five of them were hospitalized. When compared with the expected incidence, the relative risk (RR) of hospital admission for ischemic heart disease was 2.7 (95% C.I. 1.7-4.0). There were 12 deaths (4.7%) due to ischemic myocardial or sudden death (actuarial risk at 25 years: 10.2% (95% C.I. 5.3-19), compared to 2.3 cases that were expected to have died from these causes, yielding a standardized mortality ratio (SMR) of 5.3 (95% C.I. 2.7-9.3). Gender (male), pretreatment cardiac medical history and increasing age appeared to be the only significant factors for the development of ischemic heart disease. CONCLUSIONS: Despite the moderate total dose and the low (mean) fraction dose to the heart, the observed incidence of ischemic heart disease is high, especially after long follow-up periods. Treatment related cardiac disease in patients treated for Hodgkin's disease has only been reported for doses above 30 Gy. Although the optimum curative dose is still under debate, some studies recommend a dose as low as 32.5 Gy. The observed high rate of severe heart complications in this study advocates a dose reduction to this level, particularly in the regions where the coronary arteries are located. PMID- 10386716 TI - Is there more than one late radiation proctitis syndrome? AB - PURPOSE: To investigate the significance of the various late rectal symptoms that appear after radical prostatic irradiation. PATIENTS AND METHODS: Patients with localised prostate cancer treated between 1987 and 1994 at the Mater Hospital, Newcastle with radical megavoltage irradiation were recalled for examination and to complete a detailed questionnaire concerning late radiation-induced symptoms and their effects on normal daily life. The influence of patient age treatment related variables and acute proctitis symptoms occurring during therapy or the late symptoms recorded were assessed and the relationship between late symptoms and late EORTC/RTOG score and impact on normal daily life were studied. RESULTS: The presence of symptoms of acute proctitis was the only factor to predict any of three late symptoms (urgency, frequency and diarrhoea) and late EORTC/RTOG score in this series (odds ratios: 1.7-2.57, P-values: 0.009-0.0007). Cluster and discriminant function analyses revealed the presence of five subgroups of patients with varying permutations of different late rectal symptoms, including one group with minimal symptoms (P < 0.0001). While bleeding and rectal discharge were the major contributors to late EORTC/RTOG score (P < 0.0001 and 0.04), faecal urgency and bleeding were the most important factors to impact on normal daily life (P < 0.0001 and P < 0.0003). A relatively low concordance was found between late EORTC/RTOG score and the patients' self assessment on the effect of their symptoms on their normal daily lives. Some late symptoms, including bleeding and rectal discharge become less prevalent after 3 years of follow-up with a resulting improvement in EORTC/RTOG score. CONCLUSIONS: There may be more than one late (chronic) proctitis syndrome which may be linked in greater or lesser degrees to acute proctitis symptoms occurring during therapy. Urgency is a common late symptom which often has an important impact on normal daily life and deserves recognition in late normal tissue scoring systems. Assessment of the incidence of bleeding as a measure of late rectal morbidity following prostate irradiation may underestimate the impact of these chronic effects. Confirmatory studies are necessary. PMID- 10386717 TI - A correlation between residual radiation-induced DNA double-strand breaks in cultured fibroblasts and late radiotherapy reactions in breast cancer patients. AB - BACKGROUND AND PURPOSE: Prediction of late normal tissue reactions to radiotherapy would permit tailoring of dosage to each patient. Measurement of residual DNA double strand breaks using pulsed field gel electrophoresis (PFGE) shows promise in this field. The aim of this study was to test the predictive potential of PFGE in a group of retrospectively studied breast cancer patients. MATERIALS AND METHODS: Thirty nine patients, treated uniformly for breast cancer 9-15 years previously, with excision of the tumour and radiotherapy to the breast and drainage areas, were assessed clinically using the LENT SOMA scale, and a 5 mm punch biopsy taken from the buttock. Fibroblast cell strains were established and used to study residual DNA double strand breaks, using PFGE. RESULTS: There were significant correlations between the DNA assay results and the fibrosis score (r(s) = 0.46; P = 0.003), the combined fibrosis and retraction score (r(s) = 0.45, P = 0.004) and the overall LENT score (r(s) = 0.43; P = 0.006). Using polychotomous logistic regression, the fibroblast DNA assay result was an independent prognostic factor for fibrosis severity. CONCLUSIONS: There is a relationship between residual radiation-induced DNA damage in fibroblasts and the severity of the late normal tissue damage seen in the patients from whom the cells were cultured. PMID- 10386718 TI - Functional assessment of cutaneous microvasculature after radiation. AB - BACKGROUND AND PURPOSE: To determine if laser Doppler flowmetry could be used to non-invasively evaluate microvasculature function after radiation therapy (RT), we assessed blood flow response to heating in women following RT after breast conservation. MATERIALS AND METHODS: Forty women with unilateral stage I/II breast cancer treated with conservative surgery and RT were evaluated at varying intervals post RT. Ten patients were retested after an interval of 55 to 57 months to assess reproducibility of the control data. A laser Doppler probe fitted into a heat source was used to non-invasively measure blood flow in a small area of skin on the treated breast and a matched area on the untreated side. The heating element increased skin surface temperature to 40 degrees C, permitting assessment of heat stress induced changes in blood flow. RESULTS: Blood flow increased in response to heating in the untreated and treated breast skin, however the magnitude of the increase was significantly greater in the non irradiated skin. The difference in relative blood flow to the heat stress was found to be greatest in patients < or =6 months post RT. In the patients who were >36 months post RT, there was no significant difference seen in relative blood flow between the irradiated and non-irradiated sides. Cutaneous blood flow response to the heat stress was very reproducible when women were reassessed 55 to 57 months after initial testing. CONCLUSIONS: Laser Doppler flowmetry can quantify the reduced response of irradiated microvasculature to a heat stress. The difference in relative blood flow to a heat stress is greatest in patients < or =6 months post RT, and normalized in patients >36 months post RT. PMID- 10386719 TI - The benefit of Beam's eye view based 3D treatment planning for cervical cancer. AB - PURPOSE: The aim of this study was to evaluate the possibility of Beam's eye view (BEV) based three dimensional (3D) treatment planning, to reduce portions of organs at risk included in the treated volume without increasing the risk of geographical miss in external beam therapy of cervical cancer. MATERIALS AND METHODS: Three dimensional dose distribution of BEV based 3D treatment plans was compared to the 3D dose distribution derived from a four-field-box-technique using standard portals. A total of 20 patients with cervical cancer stage FIGO IIB and FIGO IIIB was included. Dose distribution in the target volumes and in the organs at risk of BEV based treatment planning, was compared to the dose distribution of the standard field technique using dose-volume-histograms. RESULTS: In 4/20 patients (20%) a geographical miss at the cervix uteri was observed for the standard field technique. The BEV based treatment planning resulted in an adequate coverage of target volume and additionally in a reduction of portions of bladder and bowel volume included in the treated volume (-13.5, 10%). In contrast the BEV based technique resulted in an increase of portions of the rectum volume included in the treated volume compared to standard portals due to a shift of the rectum by the enlarged cervix uteri from its posterior to a lateral position. An overall 7% reduction of treated volume was observed, although the maximum width of lateral fields increased for the BEV technique. Moreover, we have found a remarkable impact of bladder fillings on the amount of bowel and bladder volume included in the treated volume. CONCLUSION: BEV based 3D treatment planning for external beam therapy of cervical cancer offers a possibility to avoid geographical miss of part of the CTV with reduced portions of bladder and bowel volume included in the treated volume. PMID- 10386720 TI - Patterns of radiotherapy for early breast cancer in Northern Italy compared with European and national standards. AB - PURPOSE: To assess the current practice of early breast cancer (EBC) post operative irradiation in Northern Italy and to evaluate its conformance with European standards and recently defined national guidelines. MATERIALS AND METHODS: Fifty Radiotherapy departments in Northern Italy received a questionnaire assessing parameters on pre-treatment evaluation of patients, on preparation, prescription and execution phases of irradiation (XRT), on surgery XRT-chemotherapy integration and on follow-up. The analysis of collected information was compared with both the 1991 EORTC-EUSOMA guidelines and the 1997 AIRO (Italian Association for Radiation Oncology) minimal requirements on EBC post-operative irradiation. RESULTS: Thirty-nine out of 50 (78%) departments answered the questionnaire. All treat T1-T2 tumours, after tumourectomy or, mostly, quadrantectomy. The mean interval between surgery and XRT is 45 +/- 14 days. Chemotherapy is delivered concurrently in 70% of departments, CTV is represented by residual mammary gland in 100% of cases, while 38% and 52% of departments occasionally treat internal mammary and axillary or supra-clavicular nodes, respectively. Total dose delivered to the whole breast is 46-50 Gy in 98% (1.8-2 Gy/fraction). The tumour bed is boosted in 79% of cases. An immobilization device is used in 28% of cases CTV is clinically localized in 62% of patients. Tangential fields are simulated in 85% of centres, with film storage in 78% of cases. Co-60 units are used in 58% and/or 4-6 MV X-rays in 70% of centres, mostly utilizing beam modifiers. Computerized treatment planning is performed in 95% of cases. Fifty-five percent of departments prescribe the dose at the ICRU point. Portal films are routinely taken in 50% of cases. Boost irradiation is mainly performed using external XRT. Lastly, acute and late side effects and cosmesis are respectively evaluated in 100%, 98% and 90% of centres. CONCLUSIONS: Results on current practice in Northern Italy generally show a good conformance with European standards. However, some variables related to treatment prescription, simulation and treatment planning need to be standardized. This set of information was largely utilized by the AIRO to define national guidelines adapted to the Italian resources and situation. PMID- 10386721 TI - External audit on the clinical practice and medical decision-making at the departments of radiotherapy in Budapest and Vienna. AB - PURPOSE: To present an example of how to study and analyze the clinical practice and the quality of medical decision-making under daily routine working conditions in a radiotherapy department, with the aims of detecting deficiencies and improving the quality of patient care. METHODS: Two departments, each with a divisional organization structure and an established internal audit system, the University Clinic of Radiotherapy and Radiobiology in Vienna (Austria), and the Department of Radiotherapy at the National Institute of Oncology in Budapest (Hungary), conducted common external audits. The descriptive parameters of the external audit provided information on the auditing (auditor and serial number of the audit), the cohorts (diagnosis, referring institution, serial number and intention of radiotherapy) and the staff responsible for the treatment (division and physician). During the ongoing external audits, the qualifying parameters were (1) the sound foundation of the indication of radiotherapy, (2) conformity to the institution protocol (3), the adequacy of the choice of radiation equipment, (4) the appropriateness of the treatment plan, and the correspondence of the latter with (5) the simulation and (6) verification films. Various degrees of deviation from the treatment principles were defined and scored on the basis of the concept of Horiot et al. (Horiot JC, Schueren van der E. Johansson KA, Bernier J, Bartelink H. The program of quality assurance of the EORTC radiotherapy group. A historical overview. Radiother. Oncol. 1993,29:81-84), with some modifications. The action was regarded as adequate (score 1) in the event of no deviation or only a small deviation with presumably no alteration of the desired end-result of the treatment. A deviation adversely influencing the result of the therapy was considered a major deviation (score 3). Cases involving a minor deviation (score 2) were those only slightly affecting the therapeutic end results, with effects between those of cases with scores 1 and 3. Non-performance of the necessary radiotherapeutic procedures was penalized by the highest score of 4. Statistical evaluation was performed with the BMDP software package, using variance analysis. RESULTS: Bimonthly audits (six with a duration of 4-6 h in each institution) were carried out by three auditors from the evaluating departments; they reviewed a total of 452 cases in Department A, and 265 cases in Department B. Despite the comparable staffing and instrumental conditions, a markedly higher number (1.5 times) of new cases were treated in Department A, but with a lower quality of radiotherapy, as adequate values of qualifying parameters (1-6) were more frequent for the cases treated in Department B (85.3%, 94%, 83.4%, 28.3%, 41.9% and 81.1%) than for those in Department A (67%, 83.4%, 87.8%, 26.1%, 33.2% and 17.7%). The responsible division (including staff and instrumentation), the responsible physician and the type of the disease each exerted a highly significant effect on the quality level of the treatment. Statistical analysis revealed a positive influence of the curative (relative to the palliative/symptomatic) intention of the treatment on the level of quality, but the effect of the first radiotherapy (relative to the second or further one) was statistically significant in only one department. At the same time, the quality parameters did not vary with the referring institution, the auditing person or the serial number of the audit. CONCLUSION: The external audit relating to the provision of radiotherapeutic care proved feasible with the basic conformity and compliance of the staff and resulted in valuable information to take correction measures. PMID- 10386722 TI - Tumor size in cervix cancer and metastases. PMID- 10386723 TI - Review of the assessment of single level and multilevel arterial occlusive disease in lower limbs by duplex ultrasound. AB - The purpose of this article is to review the performance of duplex ultrasound scanning in assessing lower limb arterial disease with emphasis on patients with multisegmental occlusive lesions. Several studies have reported that duplex scanning can be as accurate as angiography to localize arterial stenoses. In spite of these promising results, there still remain some difficulties and controversies. Among them, it has been reported that multisegmental disease may affect the accuracy of duplex scanning. Indeed, some studies have indicated a lower sensitivity for detecting significant stenoses distal to severe or total occlusions. It also was demonstrated that second-order stenoses were detected with lower sensitivity compared to first-order stenoses. The main reason proposed to explain this lower sensitivity is that the highly reduced flow distal to occluded or highly stenotic segments increases the difficulty of detecting significant Doppler velocity changes in the distal or secondary stenoses. The intrinsic limitations of the peak systolic velocity ratio used as a classification criterion are presented. Finally, new and promising developments in power Doppler imaging and ultrasound contrast agents are discussed, because they may allow expansion of the capabilities of current ultrasound scanning systems and provide more accurate diagnosis of patients with multiple disease. PMID- 10386724 TI - Cerebral arteriovenous transit time (CTT): a sonographic assessment of cerebral microcirculation using ultrasound contrast agents. AB - Transcranial color-coded sonography (TCCS) has been used to investigate major brain-supplying arteries, draining veins and brain parenchyma. Here, we describe a contrast-enhanced TCCS analysis of cerebral arteriovenous transit time (cTT) as a measure of cerebral microcirculation. We evaluate its reproducibility and its correlation with clinical impairment of brain function and neuropsychological tests. A total of 27 patients with cerebral microangiopathy and 30 healthy controls were examined. CTT is defined by the time an ultrasound contrast agent requires to pass from the P2-segment of the posterior cerebral artery to the vein of Galen. This was measured by comparison of power Doppler intensity in two off line defined regions of interest. Serial intraindividual cTT measurements within several min showed a good reproducibility of this parameter. cTT was significantly longer in patients with cerebral microangiopathy than in controls (Mann-Whitney U test,p < 0.001) and related to cognitive impairment measured by the Mini-Mental-State examination. We conclude that it is a quick and reliable parameter related to increased vascular resistance of the microcirculation or a rarefaction of microvessels. Further studies are needed to show the sensitivity and specificity of cTT in the diagnosis of small vessel disease and the interference of important circulation factors, such as heart failure or blood viscosity. PMID- 10386725 TI - Clinical implications of online and off-line interobserver variability in intracoronary ultrasound-guided interventions. AB - Intracoronary ultrasound (ICUS)-guided interventions imply online decision making. We investigated the on- and off-line interobserver variability in ICUS measurements and evaluated the clinical implication of the interobserver variability for ICUS-guided interventions in 40 lesions (38 patients). On the same recorded ICUS images, an online and an off-line observer independently selected and analyzed the proximal and distal reference images, the most severe stenosis image before intervention and the minimal lumen area (MLA) image after intervention. In addition, the off-line observer analyzed the online selected images. The interobserver variability (percentage difference between the two observers) of ICUS measurements was determined on images independently selected by both observers (Analysis A) and on images selected by the online observer (Analysis B). The balloon size was determined from ICUS measurements according to the CLOUT trial. After intervention, the CLOUT and MUSIC criteria for MLA by ICUS had to be fulfilled for PTCA and stent procedures, respectively. In Analysis A, the on- and off-line interobserver variability in diameter and area measurements was maximally 9% and 18%, respectively. In Analysis B, the on- and off-line interobserver variability was maximally 6% and 11%, respectively. The off-line observer would have chosen a balloon size differing by more than 0.25 mm in 25 % (10 of 40) and 12.5% (5 of 40) of the procedures in Analysis A and B, respectively. After PTCA or stenting, the off-line observer would have taken different procedural decisions in 19% or 15% and in 13% or 7% of the procedures in Analysis A and B, respectively. IN CONCLUSION: when the same location was selected, the on- and off-line interobserver variability was low. When different locations were selected, however, the interobserver variability resulted in different balloon sizing in 25 % of the cases. Interobserver variability may be a confounding factor in the analysis of ICUS-guided interventions. PMID- 10386726 TI - Application of a semiautomatic boundary detection algorithm for the assessment of amniotic fluid quantity from ultrasound images. AB - The aim of this study was to develop a computer-assisted method to evaluate amniotic fluid volume (AFV). This was done by automatically detecting the boundaries of the amniotic fluid portion in 2-D ultrasonographic images. The study population consisted of 36 low-risk patients that were selected at random from a healthy population undergoing routine pregnancy follow-up. For each patient, images of the four quadrants of the uterus were digitized into a PC. The amniotic fluid portion in each ultrasonographic image was automatically detected, and its area was calculated. Its area was also manually determined by an expert physician (R. T.). The areas automatically detected by the algorithm were highly correlated with the areas manually delimited by the expert: r2 = 0.9722 (p < 0.01). The areas calculated by the program provide a good measure for the areas determined by the expert and may, therefore, be used for calculating the actual amniotic fluid volume. PMID- 10386727 TI - Quantitative ultrasonography of the periventricular white and grey matter of the developing brain. AB - This study addresses the value of operator-independent computer processing of ultrasonograms of the developing brain. With this aim, routine cranial ultrasonograms obtained from 39 term and preterm infants without clinical or sonographic evidence of brain damage were analyzed by five observers. The procedure, respectively, included: 1. the definition of four regions of interest (ROI), one white matter and one grey matter area on each side of the brain; 2. digitization of the sonogram data within these ROIs; 3. correction for the equipment settings, using data from a tissue-mimicking phantom as a reference; and 4. calculation of four sonogram characteristics (i.e., mean echo level, MEAN, signal-to-noise ratio, SNR, and axial and lateral correlation, CORAX and CORLAT, of the echo level co-occurrence matrix). Significant differences between both sides of the brain or a significant influence of ROI size were not found. The interobserver spread was considerable, but less than the intersubject spread. Two sonogram characteristics seemed strongly correlated in white and grey matter (CORAX and CORLAT) and another only in white matter (SNR with CORAX and CORLAT). MEAN seemed not to be correlated with any other characteristic. Furthermore, it was found that maturation equally decreases white and grey matter MEAN and, thus, hardly affects the ratio between the two. An effect on the other sonogram characteristics was only found in the white matter (i.e., an increase of SNR and a decrease of CORAX and CORLAT). Except for MEAN, the grey matter sonogram characteristics seem hardly affected by maturation. In view of these findings, we conclude that quantitative ultrasonography reveals white and grey matter maturation and, furthermore, provides a conceptional-age-independent reference (MEAN white:grey matter ratio) that might be found to facilitate the detection of pathologic brain alterations. PMID- 10386728 TI - Middle cerebral artery anatomy and characteristics of embolic signals: a dual gate computer simulation study. AB - In terms of microembolic signal (MES) detection, the anatomy of the middle cerebral artery (MCA) mainstem has only scarcely been considered. The vessel itself, however, could be at least partly responsible for the enormous variation when calculating the essential time difference (deltat) values of MES using the dual-gate technique. Therefore, we studied the time characteristics of MES in a computer simulation applying an anatomically realistic vessel and a dual-gate TCD approach. Three different MCA anatomies and two MES to blood intensities were simulated as well as two different sample volume settings. The MES length (proximal sample volume t1; distal sample volume t2) and deltat were calculated for different angles of insonation and sample volume depths. The calculations of the time characteristics of MES showed extreme variation, with only modest changes of the insonation angle (t1 4-34 ms; deltat 9-27 ms) or the sample volume depth (t1 7-27 ms; deltat 6-32 ms). The variation could be considerably reduced with modified TCD settings i.e., a shorter gate separation combined with a shorter receiver gate time in the distal sample volume (deltat with changing insonation angles 6-19 ms; deltat with changing insonation depths 13-17 ms). These results not only urge us to a cautious interpretation of the properties of single MES, but also contribute to an understanding of the marked deltat variation using the dual-gate technique. PMID- 10386729 TI - Sensitivity of color Doppler sonography: an experimental approach. AB - The purpose of this study was to estimate the size required for small vessels to become detectable with color Doppler sonography. A murine experimental tumor was examined with color Doppler sonography after injection of 1.5 mL of the contrast medium Levovist. Histologically, we measured vessel diameters inside the tumor, as well as in its direct neighborhood. With color Doppler at a transmit frequency of 7 MHz, vessels were only detected in the tumor's environment, but not inside. By histology, the 95% quantile of the vessel diameter distribution was found to be 21 microm inside the tumor, 37 microm in the underlying muscle, and 73 microm in the directly adjacent connective tissue. Vessels in the upper range of the size distribution in the muscle and connective tissue are probably detectable. Using the 95% quantile as an estimate, and correcting the values for possible shrinkage, using a factor of 1.91 reported in the literature, vessels in the 74 134 microm range may be detected under the given conditions, whereas vessels measuring 38 microm or less are inaccessible to color Doppler. PMID- 10386730 TI - The effect of hemodynamics, vessel wall compliance and hematocrit on ultrasonic Doppler power: an in vitro study. AB - Previous in vitro studies in rigid tubes under pulsatile flow conditions have reported a lack of a cyclic variation in blood echogenicity that contradicts in vivo results. To investigate whether or not these variations can be attributed to the compliance of the vessel wall, a series of in vitro experiments with compliant tubes, under pulsatile flow conditions, was performed. Two important factors that may affect the Doppler power were investigated: 1. the dependence on hematocrit and 2. the effect of the vessel wall elasticity. In the present study, it is shown that, at the low beat rates, the peak of the mean Doppler power within the flow cycle depends on the vessel wall compliance. When the vessel becomes more compliant, the peak is shifted from the early to the late systole. Additionally, there is a correlation between the power peak and hematocrit that is more evident in compliant vessels. At a higher pulsation rate of 37 beats/min, a different variation is observed. A drop in the power occurs near peak systole in compliant tube experiments and is more pronounced as the vessel becomes more constricted. The observed power drop agrees with previously reported in vivo results, but is not seen in rigid tube experiments. The results of this study suggest that proper interpretation of cyclic variations in Doppler power requires a knowledge of hemodynamic parameters, such as the modulus of elasticity of the vessel wall, propagation velocity or, possibly, the phase angle of input impedance. PMID- 10386731 TI - The progression of thrombus in an ex-vivo shunt model evaluated by intravascular ultrasound radiofrequency analysis. AB - We tested the ability of ultrasound radiofrequency (RF) signal analysis to characterize thrombus accumulation in a Dacron graft incorporated into the exteriorized arteriovenous shunt in 3 baboons with constant blood flow for 60 min. Thrombus formation was quantified by sequential measurements of 111Indium labeled platelet deposition. RF signals were acquired every 15 min at 2 sites in the graft, using a 2.9 Fr intravascular ultrasound catheter-based transducer (30 MHz) and digitized at 250 MHz in 8-bit resolution. Regions of interest were placed within a 0.5-mm perimeter adjacent to the graft wall. Integrated backscatter increased significantly (p < 0.001) with increasing platelet deposition. However, mean-to-standard deviation ratio of the RF envelope showed no significant change and the distribution pattern of the RF probability function remained constant and consistent with a Rayleigh scattering process. These results provide a basis for using RF analysis to monitor the time-course of thrombus formation. PMID- 10386732 TI - Modeling geometric artefacts in intravascular ultrasound imaging. AB - A model has been developed for estimating the geometric distortions in intravascular ultrasound (IVUS) imaging caused by the position of the ultrasound catheter within the artery. Geometric distortion causes degradation on cross sectional images of the vessel wall where, for characteristic positioning of the transducer within the vessel, a circular artery is seen on IVUS images as a noncircular vessel represented by more or less complex shapes. Artefacts, therefore, have a clinical impact on the accuracy of qualitative and quantitative intravascular analyses. The main distortions are due to the inclination and the off-centered position of the transducer within the vessel. These effects are increased by two factors: first, the point of origin of the ultrasound beam does not coincide with the rotation axis of the catheter; second, in the case of a mechanical rotating transducer, the ultrasound beam is not perpendicular to the long axis of the catheter, but has an inclination such that the transducer looks forward from the emitting point. All these parameters are taken into account in the three-dimensional (3-D) geometric model developed in this paper. The model was formulated to predict the geometric deformation for artery contour of various shapes and can model artefacts during stent implantation (Finet et al. 1998). Simulations were made for various geometric configurations and compared to in vitro and in vivo IVUS images. The model results are consistent with the experimental results. Finally, the model was used for estimating the values of the geometric parameters that cause distortions on ultrasonic images. PMID- 10386733 TI - Time-averaged mean velocity for volumetric blood flow measurements: an in vitro model validation study using physiological femoral artery flow waveforms. AB - This study assesses the accuracy of the volume flow measurement of the ATL HDI 3000 duplex ultrasound scanner using a model of the femoral arterial circulation. The beam profile of the transducer was measured, and used to identify regions of the beam where there may be poor insonation characteristics. The flow measurement accuracy was not found to be influenced by the vessel depth between 1.0 cm and 8.0 cm in a 0.7 cm diameter vessel. Overall accuracy was 3%+/-9%. Vessels in excess of 0.9 cm produced larger errors. In the model system, pulse rates between 60 bpm and 120 bpm had no significant effect on the measurement accuracy (p > 0.01). The results of this study suggest that accurate measurements of femoral arterial blood flow are possible. Further work will be required to assess the accuracy of the technique in vivo. PMID- 10386734 TI - A comparison of single- and dual-beam methods for maximum velocity estimation. AB - The purpose of this study was to compare the precision and accuracy of maximum velocity estimation when the target direction is known (a string phantom), and when the target direction is unknown (a flow model of arterial stenosis with stenoses of 0-80% by area). Maximum velocity was estimated using single- and dual beam methods. A linear-array system was used to acquire Doppler spectra from a single-beam direction. The same array was used for sequential acquisition of Doppler spectra from 2 beam directions; the velocity estimates from these were then compounded in a vector manner. The variation of estimated maximum velocity with beam-string angle over the range 40-80 degrees was 27% for conventional Doppler, 2.6% for angle correction from the edge of the array and 1.6% for the vector Doppler. In the stenosis model, for the single-beam methods, the highest frequency shift was obtained just prior to the point of minimum lumen. At this location, the variation with beam-vessel angle over the range 40-80 degrees was 35% for conventional Doppler, 7.4% for the correction factor method and 6.9% for correction from the edge of the array. For the vector method, the maximum velocity is obtained from within the poststenotic jet, the variation was 2% over the range 40-80 degrees. It is recommended that existing Doppler systems use the correction-factor method to reduce variation in measured maximum velocity. The use of the vector technique by future generations of Doppler systems may lead to angle-independent velocity estimation. PMID- 10386735 TI - Experimental investigation of the acoustic nonlinearity parameter tomography for excised pathological biological tissues. AB - The acoustic nonlinearity parameter B/A tomography is proposed as a novel imaging method in this paper. By using this method, the B/A tomography for various kinds of normal and pathological porcine tissues in vitro, including eight kinds of diseased livers, four kinds of diseased kidney and one kind of diseased spleen, are studied. Results indicate that all diseased tissues with changes in tissue composition and structural features exhibit higher grey-scale (higher B/A values) than corresponding normal tissues. In addition, the acoustic nonlinearity parameter is more sensitive to pathological states than acoustic linear parameters. To make a comparison with the acoustic nonlinearity parameter tomography, the acoustic linear parameter images, including two B-scan images for two kinds of diseased specimens, are also presented. Results of this paper show that the acoustic nonlinearity parameter tomography may be a novel method for tissue characterization in ultrasonic medical diagnosis. PMID- 10386736 TI - Wide-band acoustic spectroscopy of biological material based on a laser-induced grating technique. AB - A laser-induced transient grating technique enables fast noncontact acoustic measurements on transparent biological materials in a frequency range from tens of megahertz to 1 GHz. We have applied this method to the characterization of bovine vitreous and found high-frequency acoustic attenuation values to be close to those of water, with a quadratic dependence on frequency, in contrast to low frequency data. The potential of the technique for studying other biological materials, such as human stratum corneum, is demonstrated. PMID- 10386737 TI - Angular directivity of thermal coagulation using air-cooled direct-coupled interstitial ultrasound applicators. AB - The performance characteristics and thermal coagulation of tissue produced by directional air-cooled, direct-coupled interstitial ultrasound (US) applicators were evaluated. Prototype applicators (2.2 mm o.d.) were constructed using cylindrical transducers sectored into angular active zones of 90 degrees, 200 degrees, 270 degrees, and 360 degrees. Acoustic characterization of the applicators showed the beam output to be angularly directed from the active sector of the transducer and collimated within the axial extent. Empirical determination of the average convective heat transfer coefficient, resulting from airflow cooling the inner surface of the transducer, showed significantly high levels of transfer (> 700 W m(-2) degrees C(-1)) with a flow rate of 5.6 L min( 1). Thermal performance of the applicators was characterized through high temperature heating in vivo (porcine thigh muscle, 11 trials) and in vitro (bovine liver, 46 trials). Results demonstrated directional coagulation of tissue, with good correlation between the angular extent of the lesions and the active acoustic sector. Radial depth of coagulation with a 200 degrees applicator extended 8-17 mm, with a heating time of 1-10 min, respectively. Angular and axial lesion shape remained similar over the course of 1-10 min heating trials. Implementation of air-cooling within direct-coupled interstitial US applicators provided enhanced directivity of heating in angular and axial dimensions, and significantly increased the power handling and radial depth of tissue coagulation. PMID- 10386738 TI - A comparison of AIUM/NEMA thermal indices with calculated temperature rises for a simple third-trimester pregnancy tissue model. American Institute of Ultrasound in Medicine/National Electrical Manufacturers Association. AB - Temperature rises due to diagnostic ultrasound exposures have been calculated for a simple third-trimester pregnancy tissue model. This consisted of a layer of soft tissue representing the abdominal/uterine wall, a layer of liquid and a layer of fetal bone. The ultrasound field parameter used in the calculations was the temporal average of the square of the acoustic pressure (p2TA), measured in water but corrected for attenuation in the tissue model. The three-dimensional (3 D) distribution of p2TA was measured for five probes operating in B-mode, and four probes operating in pulsed Doppler and color flow imaging modes. The calculated temperature rises were compared to the AIUM/NEMA-defined thermal indices appropriate to third-trimester scanning. In B-mode, the ratio of calculated temperature rise to thermal index varied between 0.62 and 1.25, with calculated temperature rises as high as 1.4 degrees C. In color-flow imaging mode, this ratio varied between 1.26 and 2.45 and, in pulsed Doppler mode, between 1.46 and 2.92, with calculated temperature rises as high as 1.8 degrees C and 5.8 degrees C, respectively. These results indicate that, for scanning situations where bone is insonated through an overlying low attenuation liquid layer, the thermal index may substantially underestimate the maximum temperature rise that could occur. PMID- 10386739 TI - Intercomparison of acoustic output measurements of a diagnostic ultrasound device. AB - An intercomparison of acoustic output measurements with 9 participants from four countries was performed. A commercial diagnostic ultrasound device including a 3.5-MHz M-mode transducer was circulated for that purpose. The acoustic output of the device was measured and the temporal-average power output, peak-negative acoustic pressure, output beam intensity, spatial-peak temporal average derived intensity and arithmetic-mean acoustic-working frequency were determined. Although the majority of results were in good agreement with one another, there were also some significant deviations from the corresponding grand mean values, and the overall ranges of the participants' reported measurement results were as follows. Temporal-average power output: 3.8 mW to 9.0 mW; peak-negative acoustic pressure: 1.50 MPa to 2.86 MPa; output beam intensity: 7.3 mW/cm2 to 13.2 mW/cm2; spatial-peak temporal-average derived intensity: 72 mW/cm2 to 176 mW/cm2; arithmetic-mean acoustic-working frequency: 3.27 MHz to 3.33 MHz. The reasons for the occurrence of significantly low or high values could be identified by the participants concerned. The results show that correct ultrasonic measurements are not easy to conduct and strongly advise periodically checking the measurement tools and procedures. PMID- 10386740 TI - Does venous microemboli detection add to the interpretation of D-dimer values following orthopedic surgery? AB - The identification of risk factors for deep venous thrombosis (DVT) following orthopedic surgery remains unclear. We have investigated the relationship between plasma levels of D-dimer (DD), the presence or absence of microemboli 1 day after surgery, and the occurrence of DVT 7 days after total hip or knee replacement. The prevalence of DVT was 25 (13.3%) among 188 patients and was lower in 112 patients with DD < 2808 ng mL(-1) than in the 56 patients with higher DD levels: respectively, 8.0% vs. 21.4% (p < 0.05). D-dimer is not suitable for individual estimation of DVT risk. Microemboli were found in 112 (60%) of 186 subjects. The presence/absence or the frequency of the microemboli showed no relationship with the occurrence of DVT. Last, when evaluating the risk of DVT in orthopedic surgery, microemboli detection does not add to the interpretation of DD concentration. PMID- 10386741 TI - The feasibility of elastographic visualization of HIFU-induced thermal lesions in soft tissues. Image-guided high-intensity focused ultrasound. AB - The potential for visualizing high-intensity focused ultrasound (HIFU)-induced thermal lesions in biological soft tissues in vitro using elastography was investigated. Thermal lesions were created in rabbit paraspinal skeletal muscle in vivo. The rabbits were sacrificed 60 h following the treatment and lesioned tissues were excised. The tissues were cast in a block of clear gel and elastographic images of the lesions were acquired. Gross pathology of the tissue samples confirmed the characteristics of the lesions. PMID- 10386742 TI - The use of the Fogarty catheter in 1998. AB - Catheter-mediated thromboembolectomy has evolved over the last three decades along with changes in vascular disease patterns. As in situ arterial thrombosis has become more common both in natural arteries and prosthetic grafts, changes in mechanical thromboembolectomy have been necessary to optimize treatment strategies. A variety of devices and techniques are now available for transluminal clot removal including the original thromboembolectomy catheter, newer adherent clot catheters, graft thrombectomy catheters, thru-lumen balloon catheters and methods that involve fragmentation and removal of fresh thrombus by hydrodynamic means. PMID- 10386743 TI - Health-related quality of life and functional outcome following arterial reconstruction for limb salvage. AB - Vascular surgery outcomes have traditionally been measured by limb salvage and graft patency. However, as health care resources are rationed, the patient's functional outcome and quality of life will require assessment. The in situ saphenous vein graft has proven successful in achieving long-term limb salvage for patients with critical ischemia, with the expectation of preserving a life style and sense of well-being that would be lost with limb amputation. This study was conducted to measure functional capacity and quality of life in these patients. Seventy patients with successful in situ saphenous vein bypass grafts constructed for limb-threatening ischemia, followed for a mean of 45.6 months in a surveillance program with normal graft flow characteristics, were compared with a group of age and gender-matched controls with normal limb pressures and no history of vascular occlusive disease. A questionnaire was designed from standardized health status scales and administered to the two groups to assess symptoms, health perceptions, physical functioning and life quality. When comparing the groups of revascularized and control patients, symptoms and perceptions about their health were similar. However, the revascularized patients had significantly decreased functional capacity in their ability to walk various distances (P< or =0.005), perform household tasks (P< or =0.001) and bathe (P< or =0.001). The patient group with vascular grafts functioned as well as the controls only in activities of dressing and using the toilet. Indicators of life quality that rate independence and mobility, including the ability to procure groceries (P< or = 0.001), prepare meals (P< or =0.005) participate in social activities (P< or =0.001) and drive an automobile (P< or =0.01), were also significantly limited in the patients with successful vascular reconstructions. Although achieving long-term limb salvage and graft patency, the patients in this group of successful vascular reconstructions retain functional disabilities that require significant care. Despite these physical handicaps, these patients have a remarkably similar sense of well-being and lack of somatic complaints compared with the control group. This medical outcome study identifies the functional capacity and lifetime needs for vascular surgery patients that will provide useful data for those responsible for allocating health care resources. PMID- 10386744 TI - The blood supply of the hypoglossal nerve and its relevance to carotid endarterectomy. AB - Since the hypoglossal nerve is liable to injury during carotid endarterectomy and similar procedures, its blood supply was examined in microinjection studies of human cadavers. The nerve is supplied by arteries that arise from the ascending pharyngeal artery as it exits from the hypoglossal canal, the occipital artery as the nerve passes under its branch to the sternomastoid muscle, direct branches from the external carotid artery, and branches from the ascending pharyngeal artery just near the bifurcation of the common carotid artery. Within and close to the tongue, the nerve is supplied by branches from the lingual artery. Damage to the vessels supplying the nerve may account for some cases of hypoglossal palsy after carotid endarterectomy. Possible mechanisms are ischaemia, thermal or electrical injury from diathermy current conducted to the nerve, or intraneural haematoma from rupturing one or more of these fine vessels. PMID- 10386745 TI - Decreasing carotid endarterectomy length of stay at a university hospital. AB - In 1995, a clinical pathway for carotid endarterectomy patients was instituted at the authors' institution. The effect of this program on length of stay and patient outcomes was investigated. Records of 152 consecutive carotid endarterectomies performed by a single surgeon over a 45-month period with identical technique (general anesthesia, routine shunting, closure with a dacron patch) were reviewed. Comparison of patients treated under the pathway (n = 119) and those prior to that policy (n = 33) revealed no significant differences (P>0.05) in age, sex, co-morbid conditions, or surgical indication. No difference (P>0.05) was found for occurrence of complications, which included two fatal perioperative strokes (1.3%) and two myocardial infarctions (1.3%) (one fatal). No complications occurred after discharge and no patients required readmission to the hospital. Average length of stay was reduced from 6.0 to 3.3 days, with 78% of patients discharged within 48 h. Preoperative hospitalization decreased from 100 to 21%. A decrease in the use of preoperative arteriography from 100 to 10% was noted. The cost of vascular studies decreased from $2451 to $1228. Cost saving measures, including early discharge of stable patients, elimination of preoperative hospitalization and decreased use of arteriography, can be accomplished while maintaining acceptable complication rates following carotid endarterectomy in a university hospital setting. PMID- 10386746 TI - The significance of early postoperative duplex studies following carotid endarterectomy. AB - This study was conducted to evaluate the significance of duplex ultrasound performed soon after carotid endarterectomy. The records of patients with 150 carotid endarterectomies and postoperative duplex ultrasound within 24 h were reviewed. Eleven (7.3%) had abnormal studies with > or =50% stenosis. Two patients with abnormal studies sustained a perioperative stroke and three patients underwent reoperation for persistent lesions (P<0.0001). Preoperative and postoperative cerebral imaging studies (computed tomography (CT) or magnetic resonance imaging (MRI)) were performed on 114 patients. Seven of these demonstrated areas of infarction and all seven had abnormal duplex ultrasound studies. Twenty-six CT scans were performed with two positive for cerebral infarction in the two patients with clinical stroke. In the 88 MRI studies, five demonstrated small, subcortical focal areas of ischemia, which were clinically silent. The relationship of infarction on postoperative cerebral studies and abnormal postoperative duplex ultrasound was significant (P<0.0001). It was concluded that early postoperative duplex ultrasound studies of > or =50% stenosis demonstrate significant association with postoperative stroke or reoperation, as well as with ischemic changes on brain imaging studies. Earlier detection with intraoperative duplex would probably be more advantageous than postoperative duplex ultrasound. PMID- 10386747 TI - Screening for asymptomatic carotid stenosis in patients with peripheral vascular disease: a prospective study and risk factor analysis. AB - Screening for asymptomatic carotid artery stenosis using color flow duplex scan was performed on 186 Chinese patients with peripheral vascular disease. They consist of 121 male and 65 females, with a mean age of 70.6 years. A carotid bruit was present in 43 (23.1%) of the patients. Internal carotid artery stenosis of 70% or greater was detected in 46 patients (24.7%) including six total occlusions. Another 79 patients (42.5%) had internal carotid artery stenosis in the 30-69% range. Significant internal carotid artery stenosis was associated with age, male sex, the quantity and duration of smoking and a carotid bruit, and inversely with cholesterol, triglyceride and VLDL. Age, the number of cigarettes consumed per day, and carotid bruit were independent significant predictors of > or =70% internal carotid artery stenosis on logistic regression analysis. The degree of internal carotid artery stenosis is more severe in patients with a carotid bruit, and correlated positively with age (P<0.01), the number of cigarettes smoked (P = 0.04), and the duration of smoking (P = 0.03). Multiple linear regression analysis showed that only the age of the patient bears a significant relationship with the degree of internal carotid artery stenosis (P<0.01). There was no relationship between the degree of lower limb ischemia with carotid stenosis. The prevalence of > or =70% internal carotid artery stenosis in a high risk population with peripheral vascular disease was 24.7%. Routine duplex screening is worthwhile in this group of patients, particularly in male, elderly smokers. PMID- 10386748 TI - Subclavian artery aneurysm: an unusual manifestation of Takayasu's arteritis. AB - Complications of Takayasu's arteritis are typically ischemic in nature because of progressive arterial narrowing, with aneurysm formation occurring as a late sequela. A 30-year-old Black woman with Takayasu's arteritis presented with a progressively enlarging and tender pulsatile mass at the base of the right neck. Upper extremity pulses were intact. Chest computed tomography and aortography demonstrated a 6-cm aneurysm of the right subclavian artery, which originated at the takeoff from the innominate artery, which was also ectatic. There was no evidence of occlusive disease. An operation was performed via the median sternotomy with transverse extension into the supraclavicular area. The distal innominate artery, proximal common carotid artery and entire subclavian artery were resected and replaced with a bifurcated stretch ePTFE graft. The aneurysm was without thrombus or atherosclerosis and all vessels were extremely thick walled. Pathology revealed healed/healing nonspecific arteritis. Aneurysm formation is an unusual complication of Takayasu's arteritis. Previously reported sites of aneurysm formation include the thoracic and abdominal aorta, the innominate, carotid and superior mesenteric arteries, but not the subclavian artery. Of 28 patients enrolled in a recent clinical protocol at the National Institutes of Health with Takayasu's arteritis, none had aneurysm formation. The authors report surgical repair of a large aneurysm of the right subclavian artery in a young Black woman with Takayasu's arteritis. PMID- 10386749 TI - Femoral anastomotic aneurysms: pathogenic factors, clinical presentations and treatment. A study of 142 cases. AB - In this study, the files of 112 patients with a total of 142 femoral anastomotic aneurysms were reviewed. Eighty-five patients (76%) were initially operated upon for obstructive aorto-iliac disease, while the remaining 27 (24%) had abdominal aortic aneurysms repaired. The majority of the patients (104/112) were male and their mean age was 64.5 years (range 45-88). Ninety-three per cent of the subjects were smokers prior to the first operation and 43% continued to smoke at the time of their femoral anastomotic aneurysms operation. The mean delay between the initial surgery and the repair of the femoral anastomotic aneurysms was 74.5 months (range 1-228). The diagnosis was made because of a painless pulsatile mass (91/142), acute leg ischaemia (27/142), a painful pulsatile mass (12/142), haemorrhage (10/142), pseudo-post-phlebitic oedema (1/142) and microemboli of the toes (1/142). The operative mortality was 2.7% (3/112) of which two-thirds were patients with infected grafts. Two subgroups were distinguished: 10 patients with an infected femoral anastomotic aneurysm and 12 patients with recurrent femoral anastomotic aneurysms, 11 with a single recurrence and one with a double recurrence. In the infected group, the time to development of anastomotic aneurysm was shorter than for the group with non-infected femoral anastomotic aneurysms (41 versus 74.5 months) and the operative mortality was 20% (2/10). One patient developed a recurrent femoral anastomotic aneurysm and another was lost to follow-up. Two subsequent deaths occurred, which were unrelated to the femoral anastomotic aneurysms. In the group of recurrent femoral anastomotic aneurysms one patient was lost to follow-up and two patients died, but not as a result of recurrent femoral anastomotic aneurysms. A total of 122 cases underwent interposition of a new prosthetic segment between the proximal prosthesis and the distal artery (89 at the common femoral, 21 at the femoral profundis, eight at the superficial femoral and four at an existing femoro-popliteal graft). PMID- 10386750 TI - Arteriography in chronic renal failure: a case for carbon dioxide. AB - PURPOSE: The aim of this study was to assess the utilisation of carbon dioxide arteriography, performed with a simple injection system, as the imaging technique of choice in patients with chronic renal failure. METHODS: Patients with chronic renal impairment who required arterial imaging or intervention were recruited for carbon dioxide angiography. Demographic data were prospectively recorded and pre- and post-arteriogram renal function was quantified. Radiographic images were graded by an independent radiologist. RESULTS: Twenty-eight patients underwent renal or aorto-femoral studies with only one failure. There were no cases of contrast-induced nephropathy. Twenty-two of the films (79%) were graded as excellent or good, four as acceptable and two were considered to be poor (non diagnostic). CONCLUSIONS: This study has demonstrated that carbon dioxide angiography is a safe and clinically effective procedure in patients with chronic renal failure. PMID- 10386751 TI - Estrogen and progesterone receptors in normal and varicose saphenous veins. AB - The presence of estrogen and progesterone receptors was investigated in the walls of normal and varicose veins. Cryostat sections from the saphenous veins of 29 normal individuals, and varicose and normal vein segments of 32 patients with varicose veins, were stained with anti-estrogen or anti-progesterone receptor antibodies. Nuclear stain intensity was scored by three independent observers. Receptors to both hormones were detected in the nuclear regions of the intima and media in females and males. In the adventitia, estrogen and the progesterone receptors were found only in nuclei of the vasa vasorum. Estrogen receptor levels were lower in non-varicose segments of varicose veins compared with normal veins. In varicose segments, estrogen receptors were more abundant than in the non varicose parts of the same vein, especially in females. Similarly, progesterone receptor levels in the non-varicose portions were higher in females. These gender differences may be related to hormonal action. However, these differences may also be age related. These findings may be related to the involvement of sex hormones in varicosis, by mechanisms as yet unknown. PMID- 10386752 TI - Recurrent varicose veins: patterns of reflux and clinical severity. AB - Duplex scanning was used to determine patterns of recurrent varicose veins in 264 limbs and to relate these to clinical factors. All limbs had previously undergone sapheno-femoral ligation in the groin. A recurrent sapheno-femoral junction was present in 172 (65.2%). Incompetence was found in long or short saphenous veins in 232 limbs (87.9%), perforators in 176 (66.7%), and deep veins in 156 (59.1%). Residual long saphenous veins were present in 43.4% and 73.6% of limbs that were with and without stripped long saphenous veins, respectively. An incompetent thigh perforator was present in 14.0% and 15.3% of these two groups, respectively. Multiple sites of incompetence were observed in the majority (75.4%). In general, no particular reflux pattern in the groin was related to an increased incidence of ulceration. However, ulceration was more frequent in limbs with deep reflux to knee or below-knee levels. None of those with isolated reflux in the groin that was unrelated to the common femoral vein had ulceration. The pattern of reflux was unrelated to striping or non-striping of the long saphenous veins and the time since initial surgery. A history of deep vein thrombosis was invariably associated with some degree of deep reflux. A system of recurrent patterns in the groin is described for the purpose of surgical audit. In 15.1%, recurrence was attributed with some confidence to inadequate surgery. These results indicate that the pattern of recurrence is highly variable and often with multiple sites of incompetence. In a few instances, the pattern of recurrence was associated with specific clinical factors. A full work-up including duplex scanning is recommended. PMID- 10386753 TI - Diagnostic and surgical approach of innominate artery saddle embolus. AB - Patients with an acute arterial occlusion of the right upper extremity and absent axillary pulse should have a Doppler scan examination before a balloon catheter embolectomy is performed. If there is no arterial pulse detectable, an angiography should be performed afterwards to localize the embolus. In the case of a proximal arterial occlusion of the right arm, the authors recommend this procedure to prevent an embolus dislocation by catheter embolectomy and subsequent cerebral embolization. For direct surgical embolectomy the authors recommend a supraclavicular incision. PMID- 10386754 TI - Inflammatory pseudotumor of carotid artery: a case report. AB - Inflammatory pseudotumor is an uncommon round and spindle cell proliferative lesion of unknown etiology that occurs most commonly in the lung. But it also occurs in diverse extrapulmonary locations such as the abdomen, retroperitoneum, pelvis, heart, head and neck, upper respiratory tract, trunk, bladder and extremities. The extrapulmonary inflammatory pseudotumor is often larger, less well circumscribed and multinodular. Proximity of the tumor to vital structures or involvement of vital organs compromises the opportunity for complete resection, thus higher recurrence rates are often reported even after surgical treatment. The authors report a case of inflammatory pseudotumor originating from the common carotid artery in a 42-year-old female patient with a rapidly growing neck mass, treated by en-bloc resection of inflammatory pseudotumor and a long segment of common carotid artery followed by PTFE graft interposition. PMID- 10386755 TI - Advantages of surgical closure of patent ductus arteriosus without tube thoracostomy. AB - Between June 1994 and June 1995, 30 patients aged 6 months to 21 years (mean age, 6.38 years) underwent surgical closure of patent ductus arteriosus without tube thoracostomy. Comparison of surgical results with the results of 143 previous consecutive cases of patent ductus arteriosus performed with the conventional technique using chest tube thoracostomy revealed a statistically significant difference in postoperative temperature, need for narcotics, hospital stay and certain complications such as pneumothorax. There was no mortality and late results were excellent. It is concluded that patent ductus arteriosus closure without tube thoracostomy is feasible and can be safe, more comfortable and more cost effective than the standard procedure. PMID- 10386756 TI - Left subclavian-aortic bypass grafting in primary isolated adult coarctation. AB - In the adult patient, bypassing the coarcted segment with a tube graft has been described, among others, as a method of repair in re-do cases and in high-risk patients. Since 1992, and owing to its simplicity, it has become our elected approach in all adult cases. Twenty-two patients (mean age 22.8+/-7.18 years) with isolated aortic coarctation distal to the left subclavian artery were primarily treated with left subclavian-lower descending thoracic aorta bypass using a Hemashield woven double velour graft. There was no hospital mortality nor major postoperative complications. The patients were followed-up for a mean period of 2.36+/-1.29 years (range 1-5 years). Systolic blood pressure as well as the pressure gradient across the coarcted segment dropped significantly from 181.82+/-15.7/65.7+/-13.3 mm Hg to 124+/-13.63/7.41+/-6.49 mm Hg (P = 0.009 and 0.001). Sixteen patients (72.6%) were recorded to be symptom-free and normotensive and seven patients (31.8%) did not show any residual pressure gradient when last seen. The postoperative systolic pressure correlated positively with its preoperative value (P = 0.017) as well as with patient age (P = 0.015). Partial correlation, however, suggested that advanced age upon surgery was the determinant factor responsible for residual postoperative systemic hypertension (P = 0.007). Besides being simple, the procedure is low-risk, permits a significant drop in pressure gradient and improves systolic hypertension through an intermediate follow-up period. PMID- 10386757 TI - Aortic valvular replacement in octogenarians. Short-term and mid-term results in 140 patients. AB - Aortic valvular replacements were performed between 1986 and 1995 at Rouen University Hospital on 140 octogenarians (52 male and 88 female). Pure or predominant aortic stenosis was present in 115 patients, 25 had associated aortic stenosis and insufficiency or predominant aortic insufficiency. Significant coronary lesions were present in 42% of patients. An isolated aortic valvular replacement was performed in 74% of patients, associated with a bypass in 23% and a bioprosthesis was used in 90%. Valvular lesions were mainly caused by Monckeberg disease. Thirteen operative deaths occurred (9.3%). Functional recovery was satisfactory in 78%, mean hospital stay was 12 days. All well-known risk factors for aortic valvular replacement: age, coronary lesions, cardiac insufficiency, impaired ejection fraction and aortic insufficiency, led to an increase in operative mortality but were not statistically significant. Late mortality occurred in 28 patients, 99 patients are still alive at 4-91 months after surgery. The actuarial survival curve shows a 56.5% probability of surviving 5 years. Eighty per cent of survivors are able to live independently at home. PMID- 10386758 TI - Reinterventions for recurrent ischemic heart disease following a successful first re-do myocardial revascularization: predictors, indications and results. AB - Predictors for a reintervention following a successful first re-do surgical revascularization (CABG) were examined. Success and limitations of the reintervention procedures were evaluated. Between 3/88 and 3/95, 16.81% (302/1796) patients who had undergone a first re-do CABG surgery in the authors' center, required a reintervention. Graft angioplasty was performed in 158 (52.32%) patients and a second re-do CABG in 47.68% (n = 144). Graft angioplasty was preferred over surgery in patients aged 70 years or older (43% versus 24.3%, P<0.001) and in patients with unstable angina (55.6% versus 33.3%, NS) or a Left Ventricular Ejection Fraction (LVEF) <30% (34.8% versus 20%, P<0.05). Re-do CABG was preferred over graft angioplasty for multivessel revascularization (3+/-0.3 versus 1+/-0.6, P<0.001), proximal occlusive disease (P<0.001) and for graft disease of a longer duration (7.18+/-1.7 years versus 3+/-0.6 years, P<0.01). The independent predictors of a reintervention were (i) lack of arterial revascularization and (ii) inability to achieve a complete revascularization in a previous operation. The predictors of a failed graft angioplasty were diameter stenosis >70%, long occlusive lesions (multivariate), angulation, calcification and asymmetrical lesions (univariate). Failed graft angioplasty required a re-do CABG (n = 48: early 21, late 27), repeat graft angioplasty (n = 34: early 8, late 26) or transplant (n = 1). Recurrent symptoms following a second re-do CABG required a graft angioplasty (n = 6: early 2, late 4), a subsequent re-do CABG (n = 32) or a transplant (n = 4). Cumulative incidence of cardiac events at 1 month, and 1 and 8 years were: 20, 40.45 and 66.44% following graft angioplasty and 5.5, 10 and 56.55% following a second re-do CABG, respectively (P<0.05). Actuarial survival at 1 month and 6 years following graft angioplasty were 97.15 and 77.22%, and 94.7 and 83.26% after a second re-do CABG, respectively (NS). Re-do CABG was more effective and durable. Graft angioplasty provided a good palliation in suitable cases and also postponed the need for a high-risk surgical intervention for more favorable conditions. PMID- 10386759 TI - Influence of normothermic systemic perfusion temperature on cold myocardial protection during coronary artery bypass surgery. AB - OBJECTIVE: To determine the effect of normothermic systemic perfusion on myocardial injury when using cold cardioplegic techniques in patients undergoing coronary artery bypass surgery. METHOD: Sixty six patients with stable angina pectoris were prospectively randomized into three groups according to cardiopulmonary bypass temperature: hypothermia (28 degrees C, n = 22), moderate hypothermia (32 degrees C, n = 22) and normothermia (37 degrees C, n = 22). All patients received cold antegrade crystalloid cardioplegia and topical cooling with saline at 4 degrees C. Serum samples were collected for troponin T and I estimation preoperatively, 4 hours after removal of the aortic cross clamp, and 12, 24, 36 and 48 hours postoperatively. In addition, serial electrocardiographic studies were undertaken on days 1, 3 and 5. RESULTS: Patients were similar with regard to preoperative and intraoperative characteristics Four patients showed ECG changes typical of perioperative myocardial infarction but remained clinically well (28 degrees C, one; 32 degrees C, one; 37 degrees C, two). In the remaining 62 patients, serum troponin T increased significantly from a mean baseline value of 0.02 ng/ml to 1.5+/-0.9 ng/ml 4 hours after removal of the aortic cross-clamp (P<0.0001). Similarly, troponin I increased from 0.06 ng/ml to 0.63+/-0.47 ng/ml 12 hours after reperfusion (P<0.0001). Serum concentrations of both markers subsequently declined with time but remained higher than preoperative values at 48 hours. There were no differences between the three groups with respect to peak and cumulative serum troponin release. Normothermic cardiopulmonary bypass did not compromise the efficacy of cold myocardial protection when assessed by serum troponin concentrations in low risk patients undergoing coronary revascularization. PMID- 10386760 TI - Retrocaval passage of coronary bypass grafts to the inferior left ventricle wall. AB - Revascularization of the inferior side of the left ventricle is performed most often with aortocoronary free grafts. This article describes a technical improvement for anatomical fit and gain of length of these grafts by directing them to the right side of the heart after a passage behind the inferior vena cava. PMID- 10386761 TI - Third degree atrio-ventricular-block caused by malignant non-Hodgkin's lymphoma: an unusual indication for epicardial pacing. PMID- 10386762 TI - Pseudoxanthoma elasticum of the internal mammary artery. AB - A case of pseudoxanthoma elasticum of the left internal mammary artery from a 61 year-old male who underwent coronary artery bypass grafting is reported. Intraoperative evaluation of the left internal mammary artery revealed partial stenosis. Histologically, the stenotic portion showed pseudoxanthoma elasticum. This represents the first reported case of pseudoxanthoma elasticum in the internal mammary artery. PMID- 10386763 TI - Training vascular surgeons and protecting the patients' interests. PMID- 10386764 TI - Robust pupil center detection using a curvature algorithm. AB - Determining the pupil center is fundamental for calculating eye orientation in video-based systems. Existing techniques are error prone and not robust because eyelids, eyelashes, corneal reflections or shadows in many instances occlude the pupil. We have developed a new algorithm which utilizes curvature characteristics of the pupil boundary to eliminate these artifacts. Pupil center is computed based solely on points related to the pupil boundary. For each boundary point, a curvature value is computed. Occlusion of the boundary induces characteristic peaks in the curvature function. Curvature values for normal pupil sizes were determined and a threshold was found which together with heuristics discriminated normal from abnormal curvature. Remaining boundary points were fit with an ellipse using a least squares error criterion. The center of the ellipse is an estimate of the pupil center. This technique is robust and accurately estimates pupil center with less than 40% of the pupil boundary points visible. PMID- 10386765 TI - CT data sets surface extraction for biomechanical modeling of long bones. AB - In modelling applications such as custom-made implants design is useful to have a surface representation of the anatomy of bones rather than the voxel-based representation generated by tomography systems. A voxel-to-surface conversion process is usually done by a 2D segmentation of the images stack. However, other methods allow a direct 3D segmentation of the CT or MRI data set. In the present work, two of these methods, namely the Standard Marching Cube (SMC) and the Discretized Marching Cube (DMC) algorithms, were compared in terms of local accuracy when used to reconstruct the geometry of a human femur. The SMC method was found to be more accurate than the DMC method. The SMC method was capable of reconstructing the inner and outer geometry of a human femur with a peak error lower than 0.9 mm and an average error comparable to the pixel size (0.3 mm). However, the large number of triangles generated by the algorithm may limit its adoption in many modelling applications. The peak error of the DMC algorithm was 1.6 mm but it produced approximately 70% less triangles than the SMC method. From the results of this study, it may be concluded that three dimensional segmentation algorithms are useful not only in visualisation applications but also in the creation of geometry models. PMID- 10386767 TI - A comparative study for stepwise correlated binary regression. AB - Real-time monitored binary data are often recorded along with a large amount of associated covariates for biomedical image processing. Serially measured binary outcomes and covariates could be autocorrelated. Appropriate variable selection schemes are necessary to find a set of influential covariates on the changes in the correlated binary outcomes. Selected variables can be used as feedback information to reduce the dimension of the database. In this context, we examine the performance of the stepwise correlated binary regression. Several realistic situations of the real-time monitored binary data are considered in Monte-Carlo simulation. Results of a simulation study are discussed. PMID- 10386766 TI - Image processing techniques for quantitative analysis of skin structures. AB - Computer-based image processing and analysis techniques were developed for quantitative analysis of skin structures in color histological sections. Performance was compared with traditional non-image processing counting methods. Skin sections were stained with Masson's trichrome, hematoxylin and eosin, picrosirius red, or one of several elastin stains. The image processing software identified the top of the cellular epidermis and the dermal-epidermal junction and then calculated the volume of the cellular layer of the epidermis, epidermal thickness, and the ratio of the dermal-epidermal junction surface area to the in plane surface area. It also identified cells and collagen and calculated cellular densities and collagen densities in the papillary and reticular layers of the dermis. Attempts to computationally process elastin-stained sections to determine elastin density were unsuccessful. The described techniques were used in a preliminary study to compare mechanically stressed skin with control skin. Results showed significant differences in cellular density in the papillary dermis and collagen density in the reticular dermis for skin subjected to combined shear/compression or tension compared with an unstressed control. Measurements made with the computer technique and traditional technique showed comparable results; the mean difference in measurements for epidermal features was 5.33% while for dermal features it was 2.76%. Significance testing between control and experimental groups showed similar results, though for three of the 28 comparisons the computer method identified a significant difference while the traditional method did not. The computer method took longer to conduct than the traditional method, though with recent advances in computer hardware this time difference would be eliminated. PMID- 10386768 TI - Automatic finite element mesh generation for maxillary second premolar. AB - Developing three dimensional finite element mesh models for irregular geometric objects requires a large amount of manual efforts, hence limiting the three dimensional approach for dental structure analyses. An automatic procedure which can be used to generate a three dimensional finite element mesh for the maxillary second premolar was developed in this study. Firstly, a embedded second premolar was sliced and scanned parallel to the occlusal surface. A self-developed image processing system was employed to detect the boundaries of different materials within each section. An automatic mesh generation program was used on these boundaries to create tetrahedral elements based on moving nodes of uniform cube approach. Six mesh models of the second premolar with different element sizes using linear and quadratic elements were analyzed. Strain energy and von Mises stresses were reviewed for convergence in the crown regions. PMID- 10386769 TI - A graphic orientated data analysis system for hemodynamic research. AB - A software program to process and to extract physiological functional hemodynamics data has been developed and reported. The purpose of this software system is to process and capture cardiovascular hemodynamics and physiological functional data after data acquisition. The system utilized an interactive graphic display and script control to extract the data. With a minimum interface, it is capable of analyzing multiple channels of data and simultaneously obtaining the results. The extracted data includes global cardiovascular functional parameters and with script process the software will calculate stroke work from the pressure length relationship. The results are stored in files for further statistical analysis. The procedures are reliable and readily applicable to examine and analyze the acquired data with minimal observer bias. PMID- 10386770 TI - Changes in muscle mass and phenotype and the expression of autocrine and systemic growth factors by muscle in response to stretch and overload. AB - The study of the underlying mechanisms by which cells respond to mechanical stimuli, i.e. the link between the mechanical stimulus and gene expression, represents a new and important area in the morphological sciences. Several cell types ('mechanocytes'), e.g. osteoblasts and fibroblasts as well as smooth, cardiac and skeletal muscle cells are activated by mechanical strain and there is now mounting evidence that this involves the cytoskeleton. Muscle offers one of the best opportunities for studying this type of mechanotransduction as the mechanical activity generated by and imposed upon muscle tissue can be accurately controlled and measured in both in vitro and in vivo systems. Muscle is highly responsive to changes in functional demands. Overload leads to hypertrophy, whilst decreased load force generation and immobilisation with the muscle in the shortened position leads to atrophy. For instance it has been shown that stretch is an important mechanical signal for the production of more actin and myosin filaments and the addition of new sarcomeres in series and in parallel. This is preceded by upregulation of transcription of the appropriate genes some of which such as the myosin isoforms markedly change the muscle phenotype. Indeed, the switch in the expression induced by mechanical activity of myosin heavy chain genes which encode different molecular motors is a means via which the tissue adapts to a given type of physical activity. As far as increase in mass is concerned, our group have cloned the cDNA of a splice variant of IGF-1 that is produced by active muscle that appears to be the factor that controls local tissue repair, maintenance and remodelling. From its sequence it can be seen that it is derived from the IGF-1 gene by alternative splicing but it has different exons to the liver isoforms. It has a 52 base insert in the E domain which alters the reading frame of the 3' end. Therefore, this splice variant of IGF-1 is likely to bind to a different binding protein which exists in the interstitial tissue spaces of muscle, neuronal tissue and bone. This would be expected to localise its action as it would be unstable in the unbound form which is important as its production would not disturb the glucose homeostasis unduly. This new growth factor has been called mechano growth factor (MGF) to distinguish it from the liver IGFs which have a systemic mode of action. Although the liver is usually thought of as the source of circulating IGF-1, it has recently been shown that during exercise skeletal muscle not only produces much of the circulating IGF-1 but active musculature also utilises most of the IGF-I produced. We have cloned both an autocrine and endocrine IGF-1, both of which are upregulated in cardiac as well as skeletal muscle when subjected to overload. It has been shown that, in contrast to normal muscle, MGF is not detectable in dystrophic mdx muscles even when subjected to stretch and stretch combined with electrical stimulation. This is true for muscular dystrophies that are due to the lack of dystrophin (X-linked) and due to a laminin deficiency (autosomal), thus indicating that the dystrophin cytoskeletal complex may be involved in the mechanotransduction mechanism. When this complex is defective the necessary systemic as well as autocrine IGF-1 growth factors required for local repair are not produced and the ensuing cell death results in progressive loss of muscle mass. The discovery of the locally produced IGF-1 appears to provide the link between the mechanical stimulus and the activation of gene expression. PMID- 10386772 TI - Organisation of the chondrocyte cytoskeleton and its response to changing mechanical conditions in organ culture. AB - Articular cartilage undergoes cycles of compressive loading during joint movement, leading to its cyclical deformation and recovery. This loading is essential for chondrocytes to perform their normal function of maintenance of the extracellular matrix. Various lines of evidence suggest the involvement of the cytoskeleton in load sensing and response. The purpose of the present study is to describe the 3-dimensional (3D) architecture of the cytoskeleton of chondrocytes within their extracellular matrix, and to examine cytoskeletal responses to experimentally varied mechanical conditions. Uniformly sized explants of articular cartilage were dissected from adult rat femoral heads. Some were immediately frozen, cryosectioned and labelled for filamentous actin using phalloidin, and for the focal contact component vinculin or for vimentin by indirect immunofluorescence. Sections were examined by confocal microscopy and 3D modelling. Actin occurred in all chondrocytes, appearing as bright foci at the cell surface linked to an irregular network beneath the surface. Cell surface foci colocalised with vinculin, suggesting the presence of focal contacts between the chondrocyte and its pericellular matrix. Vimentin label occurred mainly in cells of the deep zone. It had a complex intracellular distribution, with linked networks of fibres surrounding the nucleus and beneath the plasma membrane. When cartilage explants were placed into organ culture, where in the absence of further treatments cartilage imbibes fluid from the culture medium and swells, cytoskeletal changes were observed. After 1 h in culture the vimentin cytoskeleton was disassembled, leading to diffuse labelling of cells. After a further hour in culture filamentous vimentin label reappeared in deep zone chondrocytes, and then over the next 48 h became more widespread in cells of the explants. Actin distribution was unaffected by culture. Further experiments were performed to test the effects of load on the cytoskeleton. Explants were placed in culture and immediately subjected to static uniaxial radially unconfined compressive loads of 0.5, 1, 2 or 4 MPa for 1 h using a pneumatic loading device. Loads greater than 0.5 MPa maintained the vimentin organisation over the culture period. At 0.5 MPa, the chondrocytes within the explant behaved as in free swelling culture. The rapid change in vimentin organisation probably relates to rapid swelling of the explants--under free-swelling conditions, these reached their maximum swollen size in just 15 min of culture. The chondrocytes' response to change in tissue dimensions, and thus to their relationship to their immediate environment, was to disassemble their vimentin networks. Loading probably counteracts the swelling pressure of the tissue. Overall, this work suggests that chondrocytes maintain their actin cytoskeleton and modify their vimentin cytoskeleton in response to changing mechanical conditions. PMID- 10386771 TI - Release of vasoactive substances from endothelial cells by shear stress and purinergic mechanosensory transduction. AB - The evidence for release of vasoactive substances from endothelial cells in response to shear stress caused by the viscous drag of passing fluids is reviewed and, in particular, its physiological significance both in short-term regulation of blood vessel tone and in long-term regulation of cell growth, differentiation, proliferation, and cell death in pathophysiological conditions is discussed. A new concept of purinergic mechanosensory transduction, particularly in relation to nociception, is introduced. It is proposed that distension of tubes (including ureter, vagina, salivary and bile ducts, gut) and sacs (including urinary and gall bladders, and lung) leads to release of ATP from the lining epithelium, which then acts on P2X2/3 receptors on subepithelial sensory nerves to convey information to the CNS. PMID- 10386773 TI - Regional differences in fibre type composition in the human temporalis muscle. AB - Anatomical and electromyographic studies point to regional differences in function in the human temporalis muscle. During chewing and biting the anterior portions of the muscle are in general more intensively activated and they are capable of producing larger forces than the posterior portions. It was hypothetised that this heterogeneity in function is reflected in the fibre type composition of the muscle. The composition and surface area of different fibre types in various anteroposterior portions of the temporalis muscle were investigated in 7 cadavers employing immunohistochemistry with a panel of monoclonal antibodies against different isoforms of myosin heavy chain. Pure slow muscle fibres, type I, differed strongly in number across the muscle. In the most posterior portion of the muscle there were 24% type I fibres, in the intermediate portion 57%, and in the most anterior portion 46%. The mean fibre cross-sectional area (m-fcsa) of type I fibres was 1849 microm2, which did not differ significantly across the muscle. The proportion of pure fast muscle fibres, type IIA and IIX, remained more or less constant throughout the muscle at 13% and 11% respectively; their m-fcsa was 1309 microm2 and 1206 microm2, respectively, which did not differ significantly throughout the muscle. Pure type IIB fibres were not found. The relative proportion of hybrid fibres was 31% and did not differ significantly among the muscle portions. Fibre types I + IIA and cardiac alpha + I + IIA were the most abundant hybrid fibre types. In addition, 5% of the type I fibres had an additional myosin isoform which has only recently been described by means of electrophoresis and was named Ia. In the present study they were denoted as hybrid type I + Ia muscle fibres. It is concluded that intramuscular differences in type I fibre distribution are in accordance with regional differences in muscle function. PMID- 10386774 TI - Distribution of fast myosin heavy chain-based muscle fibres in the gluteus medius of untrained horses: mismatch between antigenic and ATPase determinants. AB - The distribution of muscle fibres classified on the basis of their content of different myosin heavy chain (MHC) isoforms was analysed in muscle biopsies from the gluteus medius of adult untrained horses by correlating immunohistochemistry with specific anti-MHC monoclonal antibodies and standard myofibrillar ATPase (mATPase) histochemistry. Percutaneous needle biopsies were taken at 3 depths (20, 40 and 60 mm) from 4 4-y-old Andalusian stallions. The percentage of 'pure' I MHC fibres increased whereas that for pure IIX MHC fibres decreased from the most superficial to the deepest sampling site. Within the fast fibres, types IIA and IIAX MHC-classified fibres were proportionately more abundant in the deepest sampling site than in the superficial region of the muscle. The immunohistochemical and histochemical characterisation of a large number of single fibres (n = 1375) was compared and correlated on a fibre-to-fibre basis. The results showed that 40% of the fibres analysed were pure type I (expressing only MHC-I); they showed correct matching between their antigenic and mATPase determinants. In contrast, within the fast fibres, a considerable proportion of fibres were found showing a mismatch between their immunohistochemical and mATPase profiles. The most common mismatched fibre phenotypes comprised fibres displaying coexpression of both fast MHCs when analysed by immunocytochemistry, but showing an mATPase profile similar to typical IIX fibres (moderate mATPase reaction after preincubation at pH 4.4). Considered altogether, the total mismatched fibres represented only 4.2% of the whole fast fibre population in the superficial region of the muscle, but their proportion increased to 15.6% and 38.4% in the middle and deep regions, respectively, of gluteus medius. It is concluded that a considerable number of hybrid fast MHC IIAX fibres are present in the gluteus medius of untrained horses, suggesting that equine type II fibres have probably been misclassified in numerous previous publications based on the use of histochemistry alone. This has important implications in attempts to study the physiological properties of fast fibre types adequately in horses. PMID- 10386775 TI - The structural effect of systemic NGF treatment on permanently axotomised dorsal root ganglion cells in adult rats. AB - The effect of systemic NGF treatment on loss and shrinkage of dorsal root ganglion cells was studied in adult male rats after permanent axotomy. Nineteen 16 to 18-wk-old rats had their right 5th lumbar spinal nerve ligated and cut approximately 7 mm peripheral to the ganglion. Two days before the operation, treatment with subcutaneous injections of human recombinant NGF (1.0-0.5 mg/kg/day) was started in 9 test rats; 10 controls were given saline injections. After 1 mo the levels of substance P (SP) and calcitonin gene related peptide (CGRP) were significantly increased in intact sciatic nerve. The number and mean volume of perikarya were estimated using assumption-free stereological techniques including vertical sections, the Cavalieri principle, optical disectors, the planar rotator and systematic sampling techniques. Systemic NGF administration had no influence on survival of primary sensory neurons after axotomy. The number of perikarya was 14300 (S.D. = 1800) in axotomised ganglia in control rats versus 14700 (S.D. = 2100) in axotomised ganglia of NGF treated rats. The reduction of perikarya volume after axotomy was significantly less after NGF treatment (11600 microm3 in the control group versus 8000 microm3 in the NGF treated group). However, the apparent protection of NGF-treatment on perikaryal volume is explained by a hitherto unrecognised size effect on nonaxotomised dorsal root ganglion cells. The untreated rats had a mean volume of 24700 microm3 (S.D. = 2700 microm3) whereas rats treated with NGF had a volume of 20400 microm3 (S.D. = 1700 microm3) on the nonaxotomised side. In conclusion, systemic NGF treatment in adult rats has no effect on dorsal root ganglion cell loss in permanent axotomy whereas perikaryal size of intact nonaxotomised cells is reduced. PMID- 10386776 TI - Structure, distribution and innervation of muscle spindles in avian fast and slow skeletal muscle. AB - Muscle spindles in 2 synergistic avian skeletal muscles, the anterior (ALD) and posterior (PLD) latissimus dorsi, were studied by light and electron microscopy to determine whether morphological or quantitative differences existed between these sensory receptors. Differences were found in the density, distribution and location of muscle spindles in the 2 muscles. They also differed with respect to the morphology of their capsules and intracapsular components. The slow ALD possessed muscle spindles which were evenly distributed throughout the muscle, whereas in the fast PLD they were mainly concentrated around the single nerve entry point into the muscle. The muscle spindle index (number of spindles per gram wet muscle weight) in the ALD was more than double that of its fast-twitch PLD counterpart (130.5+/-2.0 vs 55.4+/-2.0 respectively, n = 6). The number of intrafusal fibres per spindle ranged from 1 to 8 in the ALD and 2 to 9 in the PLD, and their diameters varied from 5.0 to 16.0 microm and 4.5 to 18.5 microm, respectively. Large diameter intrafusal fibres were more frequently encountered in spindles of the PLD. Unique to the ALD was the presence of monofibre muscle spindles (12.7% of total spindles observed in ALD) which contained a solitary intrafusal fibre. In muscle spindles of both the ALD and PLD, sensory nerve endings terminated in a spiral fashion on the intrafusal fibres in their equatorial regions. Motor innervation was restricted to either juxtaequatorial or polar regions of the intrafusal fibres. Outer capsule components were extensive in polar and juxtaequatorial regions of ALD spindles, whereas inner capsule cells of PLD spindles were more numerous in juxtaequatorial and equatorial regions. Overall, muscle spindles of the PLD exhibited greater complexity with respect to the number of intrafusal fibres per spindle, range of intrafusal fibre diameters and development of their inner capsules. It is postulated that the differences in muscle spindle density and structure observed in this study reflect the function of the muscles in which they reside. PMID- 10386777 TI - Identification and localisation of glycoconjugates in the olfactory mucosa of the armadillo Chaetophractus villosus. AB - Conventional histochemistry and the binding patterns of 22 biotinylated lectins were examined for characterisation of glycoconjugates in the components of the olfactory mucosa of the armadillo Chaetophractus villosus. The mucous lining the olfactory epithelium showed binding sites for DSL, WGA, STL, LEL, PHA-E and JAC. Only the basilar processes of the supporting cells stained for Con-A and S-Con A. The olfactory receptor neurons stained with LEL, LCA, Con A, S-Con A, JAC and PNA. The layer of basal cells did not react with any of the lectins studied. Bowman's glands in the lamina propria showed subpopulations of acinar cells reacting with SBA, S-WGA, WGA, STL, Con A, PSA, PNA, SJA, VVA, JAC and S-Con A, but in our optical studies with lectins we were unable to differentiate between mucous and serous cells in the way that is possible on electron microscopy. The ducts of Bowman's glands were labelled with S-WGA, STL, LEL, PHA-E, BSL-I and JAC. This histochemical study on the glycoconjugates of the olfactory mucosa in the order Xenarthra provides a basis for further experimental investigations. PMID- 10386778 TI - The impact of mammalian reproduction on cancellous bone architecture. AB - Pregnancy and lactation make demands on maternal calcium homeostasis which may affect bone strength. Recently, changes in cancellous architecture have been described in iliac crest bone biopsies from normal pregnant women but the rarity of such human material means an animal model is essential. The microanatomy of cancellous bone was compared in uniparous and multiparous rats using undecalcified histological sections of lumbar and caudal vertebrae and also proximal femora. An automated trabecular analysis system (TAS) measured a comprehensive range of structural variables including the trabecular number, connectivity and width. In the first pregnancy cycle an early stimulation of bone formation (which quadrupled at some sites) was indicated by an increase in the skeletal uptake and spacing of double calcein labels and the immediate generation of thicker more numerous and interconnected trabeculae. A 40% increase in cancellous bone volume was observed in the lumbar spine in comparison with age matched virgin controls. In contrast, a rapid succession of 3 pregnancy cycles (including lactation) culminated in cancellous atrophy of 15% at the same site, with a loss in trabecular number ranging from 20% (caudal vertebra) to 30% (lumbar vertebrae). In comparison, the proximal femur lost 40% of its struts but, nevertheless, uniquely sustained its cancellous bone volume. When lactation was excluded the number of struts lost was halved although trabecular thinning then took place which was sufficient to maintain the previous 15% deficit in bone volume. It was concluded that a single pregnancy strengthens the cancellous component of the maternal skeleton while a quick succession of pregnancies weakens it. Lactation influences the pattern of bone loss but not its amount. PMID- 10386779 TI - The effect of the timing of ethanol exposure during early postnatal life on total number of Purkinje cells in rat cerebellum. AB - We have previously shown that exposing rats to a high dose of ethanol on postnatal d 5 can affect Purkinje cell numbers in the cerebellum whilst similar exposure on d 10 had no such effect. The question arose whether a longer period of ethanol exposure after d 10 could produce loss of Purkinje cells. We have examined this question by exposing young rats to a relatively high dose (approximately 420-430 mg/dl) of ethanol for 6 d periods between the ages of either 4 and 9 d or 10 and 15 d of age. Exposure was carried out by placing the rats in an ethanol vapour chamber for 3 h per day during the exposure period. Groups of ethanol-treated (ET), separation controls (SC) and mother-reared controls (MRC) were anaesthetised and killed when aged 30 d by perfusion with buffered 2.5% glutaraldehyde. Stereological methods were used to determine the numbers of Purkinje cells in the cerebellum of each rat. MRC, SC and rats treated with ethanol between 10-15 d of age each had, on average, about 254-258 thousand cerebellar Purkinje cells; the differences between these various groups were not statistically significant. However, the rats treated with ethanol vapour between 4-9 d of age had an average of only about 128000+/-20000 Purkinje cells per cerebellum. This value was significantly different from both the MRC and group matched SC animals. It is concluded that the period between 4 and 9 d of age is an extremely vulnerable period during which the rat cerebellar Purkinje cells are particularly susceptible to the effects of a high dose of ethanol. However, a similar level and duration of ethanol exposure commencing after 10 d of age has no significant effect on Purkinje cell numbers. PMID- 10386780 TI - Immunohistochemical evidence suggests intrinsic regulatory activity of human eccrine sweat glands. AB - Immunohistochemistry of normal eccrine sweat glands was performed on paraffin sections of human skin. Immunoreactivity (ir) for neuron specific enolase, S100 protein (S100), regulatory peptides, nitric oxide synthase type I (NOS-I) and choline-acetyltransferase (ChAT) was found in small nerve bundles close to sweat glands. In the glands, secretory cells were labelled with anticytokeratin antibody. Using antibodies to S100, calcitonin gene-related peptide (CGRP) and substance P (SP) a specific distribution pattern was found in secretory cells. Granulated (dark) and parietal (clear) cells were immunopositive for CGRP, and S100 and SP, respectively. Immunoreactivity was diffuse in the cytoplasm for CGRP and S100, and peripheral for SP. Myoepithelial cells were not labelled. Electron microscopy revealed electron dense granules, probably containing peptide, in granulated cells. Using antibodies to NOS-I and ChAT, ir was exclusively found in myoepithelial cells. Immunoreactivity for the atrial natriuretic peptide was absent in sweat glands. These results provide evidence for the presence of both regulatory peptides involved in vasodilation and key enzymes for the synthesis of nitric oxide and acetylcholine in the secretory coil of human sweat glands. It is suggested that human sweat glands are capable of some intrinsic regulation in addition to that carried out by their nerve supply. PMID- 10386782 TI - Dynamic injury tolerances for long bones of the female upper extremity. AB - This paper presents the dynamic injury tolerances for the female humerus and forearm derived from dynamic 3-point bending tests using 22 female cadaver upper extremities. Twelve female humeri were tested at an average strain rate of 3.7+/ 1.3%/s. The strain rates were chosen to be representative of those observed during upper extremity interaction with frontal and side airbags. The average moment to failure when mass scaled for the 5th centile female was 128+/-19 Nm. Using data from the in situ strain gauges during the drop tests and geometric properties obtained from pretest CT scans, an average dynamic elastic modulus for the female humerus was found to be 24.4+/-3.9 GPa. The injury tolerance for the forearm was determined from 10 female forearms tested at an average strain rate of 3.94+/-2.0%/s. Using 3 matched forearm pairs, it was determined that the forearm is 21% stronger in the supinated position (92+/-5 Nm) versus the pronated position (75+/-7 Nm). Two distinct fracture patterns were seen for the pronated and supinated groups. In the supinated position the average difference in fracture time between the radius and ulna was a negligible 0.4+/-0.3 ms. However, the pronated tests yielded an average difference in fracture time of 3.6+/-1.2 ms, with the ulna breaking before the radius in every test. This trend implies that in the pronated position, the ulna and radius are loaded independently, while in the supinated position the ulna and radius are loaded together as a combined structure. To produce a conservative injury criterion, a total of 7 female forearms were tested in the pronated position, which resulted in the forearm injury criterion of 58+/-12 Nm when scaled for the 5th centile female. It is anticipated that these data will provide injury reference values for the female forearm during driver air bag loading, and the female humerus during side air bag loading. PMID- 10386781 TI - Concerning the ultrastructural origin of large-scale swelling in articular cartilage. AB - The swelling behaviour of the general matrix of both normal and abnormally softened articular cartilage was investigated in the context of its relationship to the underlying subchondral bone, the articular surface, and with respect to the primary structural directions represented in its strongly anisotropic collagenous architecture. Swelling behaviours were compared by subjecting tissue specimens under different modes of constraint to a high swelling bathing solution of distilled water and comparing structural changes imaged at the macroscopic, microscopic and ultrastructural levels of resolution. Near zero swelling was observed in the isolated normal general matrix with minimal structural change. By contrast the similarly isolated softened general matrix exhibited large-scale swelling in both the transverse and radial directions. This difference in dimensional stability was attributed to fundamentally different levels of fibril interconnectivity between the 2 matrices. A model of structural transformation is proposed to accommodate fibrillar rearrangements associated with the large-scale swelling in the radial and transverse directions in the softened general matrix. PMID- 10386783 TI - Abnormal disposition of a branch of the ulnar nerve in the flexor retinaculum. PMID- 10386784 TI - Bilateral superficial median arteries. PMID- 10386785 TI - Cloning, functional expression and characterization of a human olfactory receptor. AB - The human olfactory system can recognize and discriminate a large number of different odorant molecules. The detection of chemically distinct odorants begins with the binding of an odorant ligand to a specific receptor protein on the olfactory neuron cell surface. To address the problem of olfactory perception at a molecular level, we have cloned, functionally expressed and characterized the first human olfactory receptor (OR 17-40). Application of a mixture of hundred different odorants elicited a transient increase in intracellular calcium at HEK 293-cells which were transfected with a plasmid containing the receptor encoding DNA and a membrane import sequence. By subdividing the odorant mixture in smaller groups we could identify a single component which represented the only effective substance: helional. Testing some structurally closely related molecules we found only one other compound which also could activate the receptor: heliotropyl acetone. All other compounds tested were completely ineffective. These findings represent the beginning of molecular understanding of odorant recognition in humans. PMID- 10386786 TI - Transregulation of erbB expression in the mouse olfactory bulb. AB - Previously, we have shown that erbB-3 expression is restricted to the ensheathing cells of the olfactory nerve layer, while erbB-4 is found in the periglomerular and mitral/tufted cells of the olfactory bulb and in cells coming out from the rostral migratory stream of the subependymal layer. In the present work, we have treated adult mice with zinc sulfate intranasal irrigation and analyzed erbB-3 and erbB-4 expression in the deafferented olfactory bulb. Following treatment, olfactory axons undergo degeneration, as indicated by the loss of OMP expression in the deafferented olfactory bulb. The thickness of the olfactory nerve layer is reduced, but the specific intensity of erbB-3 labeling in the remaining olfactory nerve layer is increased with respect to control. Interestingly, following deafferentation, erbB-4 immunoreactivity decreases specifically in cell types that normally make synaptic contacts with primary olfactory neurons in the glomeruli, i.e. periglomerular and mitral/tufted cells. Partial lesion of the olfactory epithelium allows regenerative axon growth of olfactory neurons to the olfactory bulb. Following olfactory axon regeneration, erbB-3 and erbB-4 immunoreactivity in the olfactory bulb is similar to control. Thus, like tyrosine hydroxylase, the down regulation of erbB-4 expression in the periglomerular cells is reversible. PMID- 10386787 TI - Distribution of vimentin in the developing chick taste bud during the perihatching period. AB - The tissue environment within which taste bud cells develop has not been wholly elaborated. Previous studies of taste bud development in vertebrates, including the avian chick, have suggested that taste bud cells could arise from one, or several tissue sources (e.g. crest-mesenchyme, local ectoderm or endoderm). Thus, molecular markers which are present in gemmal as well as interfacing (peribud epithelium; mesenchyme-epithelium) regions, and their degree of expression during stages of taste bud development, are of special interest. The intermediate filament protein, vimentin, occurs in mesenchymal and mesodermally-derived (e.g. endothelial, fibroblast) cells as well as highly proliferating epithelium (e.g. tumors). The present study in chick gustatory tissue utilized antibodies against vimentin and the avidin-biotin-peroxidase technique to evaluate vimentin immunoreactivity (IR) within a timeframe which includes: 1) early stages of the taste bud primordium [embryonic days (E)17-E18)]; 2) the beginning of an accelerated bud cell proliferation at the time of initial, taste bud pore opening [around E19]; 3) attaining the adult complement of taste buds [around posthatch (H) day 1], and 4) completed organogenesis (H 17). During this time span, vimentin-IR was characterized in a region including and sometimes bridging taste bud and subepithelial connective tissue, whereas non-gustatory surrounding epithelium and salivary glands were vimentin-immuno-negative. Intragemmally, the proportion of vimentin-IR cells as related to total taste bud cells peaked at E19. These results indicate that vimentin expression, in part, is related to the onset of taste bud cell proliferation and suggest that mesenchyme could be one source of taste bud cells. Secondly, fibronectin, an extracellular matrix component of the epithelial basement membrane interface with mesenchyme, was expressed at or near the apical surfaces of taste bud cells projecting into the bud lumen, and in the basal gemmal region suggesting the possible role of fibronectin as a chemotactic anchor for differentiating and migrating taste bud receptor cells. Lastly, neuron-specific enolase-IR indicates that axonal varicosities are already present intragemmally at E17-E18, that is, during the incipient period of identifiable taste bud primordia. PMID- 10386788 TI - Patch clamp recording of the responses to three bitter stimuli in mouse taste cells. AB - Although several pathways of bitter taste signal transduction have been proposed in taste cells, these mechanisms have not been elucidated in detail. To investigate the diversity of responses to bitter stimuli, we recorded the electrophysiological responses to quinine, denatonium and naringin using whole cell patch clamp technique in isolated taste cells of C57BL/6J mice. Ten mM quinine induced depolarizing response under the current clamp mode, and inward current response under the voltage-clamp mode (holding potential -80 mV) using both K+ (with pseudo intracellular solution) and Cs+ (K+ was substituted by Cs+ in the pseudo intracellular solution) pipettes. However, when the K+ pipette was used, the membrane conductance was suppressed and activated in succession. On the other hand, the membrane conductance was only activated when the Cs+ pipette was used. Half to one mM denatonium induced depolarizing response under the current clamp mode, and outward current response under the voltage clamp mode with both pipettes. Using these pipettes, the membrane conductance was activated or suppressed in the individual case. Naringin-induced responses were not detected in these measurements. These electrophysiological recordings suggest that multiple transduction mechanisms are involved in bitter taste perception in mouse taste cells. PMID- 10386789 TI - Simultaneous recordings from two physiologically different types of relay neurons, mitral cells and ruffed cells, in the olfactory bulb of goldfish. AB - Anatomical differences characterizing mitral cells and ruffed cells were published by Kosaka and Hama in three teleost species. Physiological responses from both different types of relay neurons were recorded extracellularly and simultaneously in the plexiform layer using a single tungsten microelectrode. During interstimulus intervals mitral cells responded with higher, frequently burst-like impulse rates triggered by the activity of epithelial receptor neurons. The mitral cell activity could be totally suppressed during local anesthesia of the olfactory epithelium. Ruffed cell impulse rates were low, and each action potential triggered a long-lasting (3-5 ms), continuously variable, summed up granule cell potential. In contrast to mitral cells, blockade of epithelial receptor cells significantly increased the activity of ruffed cells. I.e., the ruffed cells, which have no input from the olfactory epithelium, are spontaneously active, and are laterally inhibited by granule cells activated by mitral cells. During olfactory stimulation contrasting interactions between mitral cells and ruffed cells resulting in a drastic intensification of centrally transmitted information, frequently were recorded. An excitation of mitral cells activity via granule cells laterally inhibited the ruffed cells activity, and an inhibition of mitral cells activity simultaneously "released" an excitation of ruffed cells. This is the first physiological determination of different types of relay neurons in the olfactory bulb of fish. PMID- 10386790 TI - Selective and reversible blockage of a fatty acid odour response in the olfactory bulb of the frog (Rana temporaria). AB - The lectin concanavalin A (ConA) when applied to the olfactory mucosa (OM) of frog and rat, is reported to partially inhibit electro-olfactogram (EOG) responses to fatty acid odours. Control odours like isoamyl acetate were not affected. We have now studied in the frog whether this treatment affects the corresponding olfactory bulb (OB) response. The OB surface was impregnated with a voltage-sensitive dye (RH 414). Spatial and temporal patterns of odour response were measured by changes in dye fluorescence that occur when OB neurons fire. The apparatus, consisted of an epi-fluorescent microscope coupled to a 64 x 64 pixel CCD photodetection camera. This allowed imaging over an 0.9 mm2 area of the OB glomerular layer to high resolution. When the frog OM was bathed with 5 mg ml(-1) ConA in Ringer's solution, the n-butyric acid odour response in the OB largely disappeared while the isoamyl acetate response did not change. When this experiment was repeated in the presence of 20 mM methyl alpha-D mannopyranoside (a ConA inhibitor), ConA failed to inhibit the n-butyric acid response. Moreover the ConA effect was partially reversible. A Ringer's wash of the OM after ConA treatment, partially restored the OB response to n-butyric acid. Thus the olfactory bulb results seem compatible with the EOG results and reinforce the notion that ConA selectively prevents n-butyric acid sensitive olfactory receptor neurons from firing. Chemical modification of the OM and their effect on OB response patterns may provide a useful approach to investigate olfactory quality coding. PMID- 10386791 TI - Overexpression, purification and photoaffinity labeling with a 3H-analogue of norfloxacin, of the GyrA and GyrB subunits of the DNA gyrase. AB - In spite of much work on DNA gyrase and quinolones for many years, our knowledge of the molecular basis of quinolone-gyrase action is still incomplete. We designed a photoaffinity labeling reagent for the quinolone target, and synthesized a norfloxacin analogue with an azide function which, under UV irradiation, becomes covalently linked to its target. For that, a large amount of purified gyrase was needed. Both subunits were purified using exclusion and affinity chromatography. A plasmid was used that allowed the overproduction of GyrA as a fusion-protein with six histidine residues at its carboxy-terminal domain. GyrA-(His)6 was purified after chromatography on a nickel-containing column, and native GyrB after chromatography on immobilized novobiocin. Reconstituted DNA gyrase (A2B2) had supercoiling activity. Photoaffinity labeling showed covalent binding of the 3H-photoaffinity analogue of norfloxacin to the gyrase-DNA complex, and mainly to the GyrA. The specific binding site remains to be explored. PMID- 10386792 TI - Pressure-sensitivity of endoplasmic reticulum membrane and nucleolus as revealed by electron microscopy. AB - Murine erythroleukemia cells submitted to high hydrostatic pressure (up to 110 MPa, 17 min. at room temperature) remain viable (Mentre et al., 1997) but present, at the ultrastructure level, perturbations which are documented in this work. In cells submitted to 50 MPa, endoplasmic reticulum membranes displayed a characteristic rigid aspect, whereas plasma membrane remained unaffected up to 110 MPa. This result is in agreement with: i) the liquid-crystalline --> gel state transition undergone by phospholipid bilayers under pressure, which involves the structuration of water at the polar-apolar interface; ii) the low concentration in cholesterol of endoplasmic reticulum membranes compared to plasma membrane, and iii) the ability of cholesterol to protect membranes against the effects of high hydrostatic pressure. Nucleoli displayed a remarkable compact aspect above 80 MPa, involving the disappearance of vacuoles and the diminution of fibrillar component, but the retention of granular component. Pressure inhibition of translation is advanced as a possible cause of this perturbation. PMID- 10386793 TI - Population pharmacokinetics of methotrexate in the guinea pig. AB - The population pharmacokinetics of an antitumoral and antiinflammatory agent, methotrexate (MTX), a folic acid antagonist, was studied in guinea pigs. Animals received an acute intraperitoneal injection of 0.25, 1 or 5 mg/kg MTX. Blood sampling was carried out for 12 hrs. after MTX administration and plasma drug concentrations were measured by fluorescence polarization immunoassay. The pharmacokinetic (PK) parameters were computed using the bayesian population model. MTX reached the level of detection at 3 hrs. for the animals injected with the lowest dose (0.25 mg/kg), at 3.5 hrs. for those animals which had the intermediate dose (1 mg/kg) and more than 6 hrs. for animals having received the highest dose (5 mg/kg). Each kinetic parameter (half life, total clearance - CLt, volume of distribution at steady state - VDSS, mean residence time - MRT - and area under curve - AUC) didn't show any significant difference between doses. MTX kinetic was linear for the first two doses (0.25 and 1 mg/kg MTX) and non-linear thereafter. MTX presented a one compartment distribution. PMID- 10386794 TI - Clinical trials for piriformis syndrome. PMID- 10386795 TI - Fact, myth, or common sense: anterior cruciate ligament reconstruction graft selection. PMID- 10386796 TI - Clinical application of femoral neck fracture fixation technique. PMID- 10386797 TI - Component removal is not mandatory in the presence of periacetabular osteolysis. PMID- 10386798 TI - Periacetabular osteolysis demands component removal. PMID- 10386799 TI - In vitro comparison of elongation of the anterior cruciate ligament and single- and dual-tunnel anterior cruciate ligament reconstructions. AB - This study evaluated strain in the normal anterior cruciate ligament (ACL) and compared it to four different double-strand hamstring tendon reconstructive techniques. Seventeen fresh-frozen knees from 11 cadavers were tested. The strain in the anteromedial and posterolateral bands of the native ACL and their equivalents in four autograft techniques were measured using differential variable reluctance transducers. The anteromedial band of the intact ACL shortened from 0 degree -30 degrees of flexion, then lengthened to 120 degrees; the posterolateral band of the intact ACL shortened from 0 degree - 120 degrees of flexion. Following ACL excision, these knees underwent reconstruction with double-strand hamstring tendons with either single tibial and femoral tunnels, single tibial and dual femoral tunnels, dual tibial and single femoral tunnels, or dual tibial and dual femoral tunnels. With the exception of the dual-band, dual-tunnel technique, all of the procedures placed greater strain on the reconstructive tissues than was observed on the native ACL, after approximately 30 degrees of flexion. These results indicate that dual-band hamstring tendon reconstructions placed with single tibial and femoral tunnels do not address the complexity of the entire ACL. Rather, these procedures appear to only duplicate the effect of the anteromedial band, while perhaps overconstraining the joint as a result of its inability to reproduce the function of the posterolateral band. During rehabilitation following ACL reconstruction, therefore, only from 0 degree - 30 degrees of the graft tissues are not significantly strained. Dual tibial and femoral tunnel techniques should be evaluated further to more closely recreate knee kinematics following ACL reconstruction. PMID- 10386800 TI - Comparison of bone-patellar tendon-bone interference screw fixation and hamstring transfemoral screw fixation in anterior cruciate ligament reconstruction. AB - While bone-patellar tendon-bone (BPTB) interference screw anterior cruciate ligament (ACL) reconstruction is a biomechanically sound construct, alternative techniques have been developed secondary to potential donor site morbidity. This study evaluates a system designed to address this problem that involves a transfemoral screw fixation device and stapling of hamstring tendons. Seven pairs of cadaveric knees underwent ACL reconstruction using either BPTB interference screw technique or semitendinosus gracilis (STG) transfemoral screw fixation and stapling. Tensile testing was performed. There was no significant difference between the two fixation types with regard to stiffness, maximum load to failure, elongation, energy to failure and yield load, displacement, and energy. PMID- 10386801 TI - Use of autologous grafts for reconstruction of osteochondral defects of the knee. AB - This study reports on 13 patients (mean age: 31 years) with a femoral condyle defect >1.5 cm2 who underwent treatment with an osteochondral graft of the same size obtained from the superior aspect of the lateral condyle, preserving the patellar groove. Mean follow-up was 61.5 months (range: 13-141 months). Twelve results were rated clinically as satisfactory with patients able to resume their normal pre-injury level of activity, and 1 case was rated as poor. No patient reported any patellar problems. Radiographic and computed tomographic evaluation demonstrated good integration of the graft in the host bone. The results of this technique at relatively long-term follow-up are encouraging, with a high percentage of subjective satisfaction. This technique appears to be reliable and provides a valid solution for treatment of wide cartilage defects when other techniques are too complex or inadequate. PMID- 10386802 TI - Sterilization of contaminated bone-tendon autografts using 10% povidone-iodine solution. AB - This study evaluates methods of sterilizing contaminated bone-tendon autografts using 10% povidone-iodine solution. Sterile grafts were prepared from human cadavers. Grafts were immersed in a suspension of either Staphylococcus aureus or Pseudomonas aeruginosa, and three sets of sterilization experiments were performed in 10% povidone-iodine for 30 minutes: one each with S. aureus and P. aeruginosa by static soaking and a third with S. aureus by serial washing with agitation. Of grafts inoculated with S. aureus, six of six grew the test organism after soaking at room temperature, as did five of six after soaking at 36 degrees C and also eight of nine after washing with agitation. Of grafts inoculated with P. aeruginosa, five of six grew the test strain after soaking at room temperature, as did six of six after soaking at 36 degrees C. Thirty minutes of exposure to aqueous 10% povidone-iodine does not adequately sterilize an inoculated graft. PMID- 10386803 TI - Elbow ligament strain under valgus load: a biomechanical study. AB - This study assessed the importance of the anterior and posterior bundles of the medial collateral ligament in the elbow by measuring in situ strain in response to valgus loads at three positions of forearm rotation throughout a full arc of motion. Strain in the anterior bundle was significantly greater than in the posterior bundle and increased with more flexion. The anterior bundle developed strain at a lower flexion angle (30 degrees) than the posterior bundle (60 degrees). Strain ratio increased with load increase for all flexion angles. Forearm position minimally affected strain. These results indicate that the anterior bundle is important in resisting a valgus load, particularly in mid flexion, while the importance of the posterior bundle increases as the elbow approaches full flexion. PMID- 10386804 TI - Periarticular neural elements in the shoulder joint. AB - Due to its unconstrained nature, the glenohumeral joint must necessarily have several mechanisms to regulate its position in space. The neural mechanisms associated with this positioning have not been fully evaluated anatomically. In this study, three fresh-frozen human cadaveric adult shoulders were dissected. Specimens were excised from the proximal biceps insertion, the superior, middle, and inferior glenohumeral ligaments, and the capsule superior to the glenohumeral ligaments. In two specimens, a portion of glenoid labrum was analyzed using a modified gold chloride staining method and light microscopy. A portion of mid biceps tendon was used as a control. In the superior glenohumeral ligament, 45% of sections contained neural elements consisting of Golgi's, Ruffini's, and Pacini's corpuscles as well as free nerve endings. The predominant types were Ruffini's and Golgi's. The middle glenohumeral ligament sections revealed all four receptor types in 42%, with the most common elements being Pacini's and Ruffini's receptors. The inferior glenohumeral ligament specimens contained the four receptor types in 48% of sections, with Ruffini's, Pacini's, and Golgi's types equally distributed. The shoulder capsule specimens revealed Ruffini's and Pacini's receptors in 47.5% of sections. Only free nerve endings were identified in the biceps tendon and glenoid labral tissue. These findings suggest that the pattern of neural elements does not appear to be random in nature and may have some correlation with the specific functions of some of the glenohumeral ligaments. PMID- 10386805 TI - Throwing-induced humeral shaft fracture in skeletally immature adolescents. PMID- 10386806 TI - Achilles tendon injury in a professional basketball player. PMID- 10386807 TI - Large autogenous osteochondral graft for replacing knee cartilage defect. PMID- 10386808 TI - Anterior cruciate ligament (ACL) ganglion cyst. PMID- 10386809 TI - Cellular and molecular causes of male infertility in spinal cord injury. PMID- 10386810 TI - Bioethics and Law Forum. PMID- 10386811 TI - Swimming upstream: views on the ethics of preconception gender selection. PMID- 10386812 TI - Detection of alpha-fetoprotein mRNA in seminoma. AB - The classic testicular tumor marker alpha-fetoprotein (AFP) is associated with nonseminomatous germ cell tumors, including embryonal carcinoma, yolk sac tumor, and teratoma. AFP is not considered to be produced by pure seminoma. However, postmortem studies have demonstrated that 30 to 45% of patients who died of seminoma initially diagnosed harbored nonseminomatous metastases and had an elevated serum AFP. We analyzed AFP expression by immunohistochemistry and by nested reverse transcription-polymerase chain reaction (RT-PCR) in 10 seminomas, 3 embryonal carcinomas, and 1 immature teratoma, diagnosed by traditional clinical methods. Positive immunohistochemical staining was observed in all embryonal carcinomas and in the teratoma but not in the seminomas. AFP mRNA, however, was found in 6 of 10 seminomas, in all embryonal carcinomas, and in the teratoma. The nucleotide sequence of PCR products was identical with that of the AFP gene. We conclude that the analysis of AFP gene expression by nested RT-PCR would be useful for detecting minute quantities of nonseminomatous germ cell elements in classic seminoma. Moreover, the existence of AFP mRNA suggests the possibility that seminoma cells can differentiate into nonseminomatous germ cells. PMID- 10386813 TI - Evidence for catecholaminergic, neuronlike cells in the adult human testis: changes associated with testicular pathologies. AB - Neuronlike, catecholaminergic cells expressing tyrosine-hydroxylase (TH) have recently been found in the testis of a nonhuman primate species, the rhesus monkey. We examined whether neuronlike cells are present in the human testis. To this end, we first determined if the genes for TH and for a voltage-activated sodium channel (NaCh), a prerequisite for neuronal excitability, are expressed in normal adult testes. Using an RT-PCR approach, cDNA clones, identical to the sequences of human TH and to the alpha subunit of a NaCh type, were isolated. Immunohistochemical methods localized the corresponding proteins in testicular biopsies from adult men (age range, 28-44 years) without testicular pathologies and from infertile patients with either Sertoli cell only (SCO) syndrome or severe hypospermatogenesis and germ cell arrest (GA). TH and NaCh antibodies, as well as antibodies recognizing dopamine-transporter protein, identified immunoreactive cells of mainly bipolar or occasionally multipolar, elongated phenotype in most, but not all, biopsies of each group (12 out of 23). The results were corroborated by identification of TH gene expression by RT-PCR approaches in biopsies. Immunoreactive cell bodies, as well as nerve fibers, were more readily detected in SCO and GA biopsies. This was quantified after immunohistochemically visualizing all testicular neuronal elements, cell bodies, and fibers, with a neurofilament 200 (NF-200) monoclonal antibody in one set of randomly selected sections from all biopsies. We found significantly increased NF 200-immunoreactive cell bodies and fibers in SCO-syndrome and GA biopsies. These results show the existence of an as yet unknown testicular catecholaminergic neuronlike cell type in the human testis. This cell type may complement and act in concert with the well-known testicular sympathetic innervation. The increase of both "intrinsic" (neuronal cells) and "extrinsic" (nerve fibers) neuronal elements in pathological testicular biopsies suggests that the two parts of the human testicular nervous system may be involved in pathogenesis and/or maintenance of GA and SCO syndromes. PMID- 10386814 TI - Measurement of prostate-specific antigen and human glandular kallikrein 2 in different body fluids. AB - It has been demonstrated that prostate-specific antigen (PSA), in spite of its name, can be detected in body fluids and tumors from a variety of organs. Investigations have shown that human glandular kallikrein 2 (hK2), a related prostate-secreted protease, can activate the zymogen form of PSA, suggesting that the two enzymes might work as a functional unit, with hK2 as the activator molecule and PSA as the effector molecule. If this is true, then hK2 should be found together with PSA in body fluids other than seminal plasma, as well. Recently, a sensitive and specific assay was devised for hK2, enabling its measurement in picogram quantities. With this assay, the concentration of hK2 was determined in samples of seminal plasma, amniotic fluid, breast milk, and saliva. Simultaneously, the samples were assayed for molecular forms of PSA. In seminal plasma, the mean PSA concentration was 0.82 mg/ml, while the hK2 level was around two orders of magnitude lower: mean value, 6.4 microg/ml. Approximately the same ratio of PSA to hK2 as in seminal plasma was found in amniotic fluid and breast milk, but in most samples, the hK2 values were too low for direct measurements and had to be concentrated prior to analysis. Measurable levels of PSA, all in the free form, were detected in amniotic fluid at the thirteenth week of gestation and then gradually increased to levels around and over 1 microg/L from the twentieth week. Significant levels of PSA were detected in amniotic fluid collected at delivery, also. Measurable levels of mammary PSA were primarily detected in colostrum, with a range from less than 0.03 microg/L to 2.1 mg/L. Around half of the molecules were in complex with protease inhibitor. Most surprisingly, determinations on saliva samples showed that none of them had detectable PSA levels but had measurable concentrations of hK2 with a mean value, 0.09 microg/L. The presence in saliva suggests that hK2 can be the human equivalent to one of the mouse salivary kallikreins with important biological function, like the epidermal growth factor-binding protein or the gamma subunit of nerve growth factor. However, this was ruled out, as a phylogenetic analysis showed that the human and mouse glandular kallikreins evolved independently from a common precursor after the separation of the primate and rodent lineages. In conclusion, the measurements show that in addition to the previously known secretion in seminal plasma, hK2 is secreted in amniotic fluid, breast milk, and saliva. Furthermore, the concerted expression of PSA and hK2 in seminal plasma, amniotic fluid, and breast milk suggests that the two proteases might form a functional unit but not always as demonstrated by the sole presence of hK2 in saliva. PMID- 10386815 TI - The effects of aging on the seminiferous epithelium and the blood-testis barrier of the Brown Norway rat. AB - Steroidogenesis and spermatogenesis decrease in aging Brown Norway rats. We therefore hypothesized that there must be accompanying morphological changes taking place in the seminiferous tubules of the aging testis. The testes of Brown Norway rats ranging in age from 3 to 24 months were prepared for light and electron microscopy. To assess the integrity of the blood-testis barrier with age, a lanthanum nitrate study was done. The normal seminiferous tubules present in rats at 3 and 12 months of age were largely replaced at 24 months by fully regressed tubules that were virtually devoid of germ cells and contained large intercellular spaces. An electron-microscopic study of these regressed tubules showed a complete loss of cyclical variations of the organelles of the Sertoli cells. The nucleus was more irregularly shaped and was present at various levels in the epithelium. The endoplasmic reticulum was a loose, vesiculated network that was unlike the elaborate, tubular, anastomotic network noted in young animals. The lysosomes were large, oddly-shaped, and contained lipidic inclusions, in contrast to the distinct membrane-bound lysosomes and dense core bodies found in the young animals. Adjacent Sertoli cell processes encompassed large, empty intercellular spaces, possibly occupied previously by germ cells. The typical Sertoli-Sertoli junctions of the blood-testis barrier in the young animal were rarely seen at 24 months and were replaced by focal contact points, usually between three Sertoli cell processes. In the aged animals, lanthanum nitrate permeated the basal and adluminal compartments, extending between Sertoli cell processes and entering the intercellular spaces and lumen. In summary, during aging, there is a breakdown of the blood-testis barrier, and there are striking changes in the appearance of Sertoli cells. These results suggest a possible intrinsic limitation that prevents stem cells from renewing themselves, whether because of a degeneration of immunological origin or because of a lack of Sertoli cell support. PMID- 10386816 TI - Pituitary adenylate cyclase-activating polypeptide (PACAP): effects on blood flow in the testis and caput epididymidis of the rat. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) is synthesized in developing germ cells in the testis and may act as a paracrine modulator of spermatogenesis and/or participate in tubule-interstitial interactions. Despite the abundance of PACAP in the organ, its role in testicular function has not yet been studied in vivo. Using laser Doppler flowmetry, the effects of PACAP on blood flow in the testis and caput epididymidis were studied on anesthetized adult rats. When given intratesticularly as 5- and 50-ng doses, PACAP increased blood flow by 55+/-21% (mean +/- SEM, P < 0.05) and by 68+/-11% at 5 mm from the injection site, respectively. Whereas 5 ng PACAP did not influence blood flow 15 mm from the site of injection, flow was reduced (-7+/-3; P < 0.05) at this site following treatment with 50 ng. Injection of 50 ng PACAP into the caput epididymidis increased epididymal blood flow by 18+/-4% (P < 0.05) at 1 mm from the injection site. None of the treatments above significantly affected the mean arterial blood pressure. Using immunohistochemistry, PACAP was observed in elongated spermatids and in the acrosomes of round spermatids in some, but not all, seminiferous tubules. Also, distinct PACAP immunoreactivity was seen in epithelial cells, particularly in clear cells, of the caput epididymidis. In conclusion, PACAP can induce vasodilatation in both testicular and epididymal microvessels and may be involved in regulating blood flow in these organs. Whereas the vasodilatory effect of PACAP is strong in the testis, the epididymal response appears to be more moderate. PMID- 10386817 TI - TGF-beta could be involved in paracrine actions in the epididymis of the marmoset monkey (Callithrix jacchus). AB - The transforming growth factor-beta1 (TGF-beta1) and the transforming growth factor-beta receptor type II (TGF-betaRII) were studied in the epididymis of sexually mature marmoset monkeys (Callithrix jacchus) by immunohistochemical localization of the protein and by polymerase chain reaction (PCR) analysis of the mRNA level. In order to specify reactive cell types, the morphology of all three segments (caput, corpus, and cauda epididymidis) was evaluated by light microscopy. Six different cell types could be distinguished: principal, basal, apical, and clear cells, as well as intraepithelial lymphocytes and macrophages. Using immunohistochemistry, specific staining for TGF-beta1 in the caput was found in 47% of the apical cells, whereas the TGF-betaRII was located in the apical portion of 91% of all principal cells. In the corpus epididymidis, 20% of the apical cells were immunopositive for TGF-beta, and binding of the receptor antibody occurred in 17% of the principal cells (all numbers based on counts of counterstained nuclei). All differences between percentages in the caput and corpus were significant as determined by chi-square test. PCR analysis revealed detectable levels of TGF-beta1 mRNA in the marmoset epididymis. Our results indicate for the first time that TGF-beta1 is synthesized in the marmoset epididymis, possibly in a different subpopulation of epididymal cells than the TGF-beta receptor type II. Thus, TGF-beta might be of functional relevance in the primate epididymis. PMID- 10386818 TI - Cloning and characterization of an androgen-dependent acidic epididymal glycoprotein/CRISP1-like protein from the monkey. AB - A cDNA encoding an acidic epididymal glycoprotein (AEG)-like, CRISP1 (cysteine rich secretory protein) protein from the monkey (Macaca mullata) epididymis has been cloned and sequenced. The monkey AEG (mAEG) has an open reading frame that encodes a protein containing 249 amino acids with a deduced molecular mass of 28 kDa. The mAEG protein sequence is 85% identical to human and 44% identical to mouse CRISP1, including all 16 conserved cysteine residues. mAEG also shows a significant amino acid homology with other CRISP proteins, rat AEG/DE, human TPX1/CRISP2, and guinea pig acrosomal autoantigen 1 (AA1). In addition, mAEG shows somewhat less homology to a toxin from the Mexican beaded lizard and to a human glioma pathogenesis-related protein. Northern blot analysis shows that the mRNA for mAEG is expressed in all the regions of the epididymis except the caput and was not detected in the testis, prostate, seminal vesicle, and brain. In castrated animals, mAEG gene expression in the epididymis is significantly diminished; however, testosterone enanthate replacement restored the normal level of expression, demonstrating that expression of mAEG is androgen dependent. Western blot analysis of monkey epididymal regions using mouse antirecombinant human AEG identified a 28-kDa protein only in the caudal region. Immunohistochemical analysis identified mAEG only in the principal cells of the cauda epididymal epithelium. Immunofluorescence analysis identified mAEG on the principal piece of the sperm tail and as small patches over the middle piece and head regions. The results described in the present study suggest that mAEG (CRISP1) is secreted in the monkey epididymis, regulated by androgens and present on epididymal spermatozoa. PMID- 10386819 TI - Three-generation evaluation of Y-chromosome microdeletion. AB - Sperm cells can be retrieved directly from the testis (testicular sperm extraction [TESE] procedure) and used for intracytoplasmic sperm injection (ICSI), circumventing underlying spermatogenetic defects. Thus, it is important that added information be available on the genetic defects in men undergoing TESE for the ICSI procedure and on the transmission of genetic factors associated with infertility to the offspring. We report a three-generation genetic analysis of a family with a case of male factor infertility. The proband, previously diagnosed as infertile, was physically examined and laboratory tested for gonadotrophic hormones, semen analysis, karyotype and Y-chromosome microdeletion screening in the blood and testis. The Y-chromosome microdeletion screening was performed by multiplex polymerase chain reaction with 20 Y-chromosome sequenced, tagged sites located at the Y chromosome. A microdeletion including the AZF-c region was detected in the azoospermic patient. His father, four brothers, and three offspring born after ICSI also underwent Y-chromosome microdeletion screening. The genetic analysis of the male members of the patient's family did not reveal similar microdeletions. The newborn male was found to bear a Y-chromosome microdeletion similar to that of his father. The fertilization capacity of the proband testicular microdeleted spermatozoa by the ICSI procedure is described. The transfer of the genetic defect raises the possibility that the son will have the same fertility problem as his father. PMID- 10386820 TI - Follicle-stimulating hormone and testosterone stimulation of immature and mature Sertoli cells in vitro: inhibin and N-cadherin levels and round spermatid binding. AB - The in vitro response of Sertoli cells isolated from adult rat testes to testosterone (T) and follicle-stimulating hormone (FSH) treatment was investigated. Sertoli cells from >70-day-old Sprague-Dawley rats were isolated by a combined enzymatic treatment followed by the removal of the majority of contaminating germ cells with immobilized peanut agglutinin lectin. Sertoli cells were then cultured for 6-10 days, forming a confluent layer with a cell viability of >83% and 74-77% purity. The contaminating cells were peritubular cells (4-6%), pachytene spermatocytes (4-5%), round spermatids (<2%), elongated spermatids (<1%), and degenerating germ cells (14.8%). The proportion of degenerating germ cells decreased from 14.8% to 8.6% between days 6 and 10 in culture. After a prestimulation culture period of 4 days, FSH treatment over a 2-day period resulted in a dose-related increase of inhibin with a median effective dose (ED50) value of 36.7+/-20.4 ng/ml in comparison with an ED50 value of 4.4+/-0.9 ng/ml obtained with immature Sertoli cell cultures from 20-day-old rats. Mature Sertoli cells, in contrast to immature Sertoli cells, were unresponsive to combined FSH + T treatment for the production of the cell adhesion protein N cadherin. FSH treatment promoted the in vitro binding of round spermatids isolated from adult testis to adult Sertoli cells in coculture. It is concluded that mature Sertoli cells in culture are responsive to FSH in terms of inhibin production and round-spermatid binding. The lack of an FSH + T-induced increase in N-cadherin or round spermatid binding is attributed to either a reduced sensitivity, or an alteration in the regulation of mature Sertoli cells by FSH + T. PMID- 10386821 TI - A lower dosage levonorgestrel and testosterone combination effectively suppresses spermatogenesis and circulating gonadotropin levels with fewer metabolic effects than higher dosage combinations. AB - Studies using exogenous high-dosage testosterone (T) or a combination regimen of physiologic T plus high-dosage levonorgestrel (LNG) administration in normal men have shown that oligoazoospermia (<3 million/mL) or azoospermia can be achieved in the majority of the men. However, these hormonal regimens have been associated with significant weight gain and suppression of serum high-density lipoprotein (HDL) cholesterol levels. We hypothesized that a combination of physiologic exogenous testosterone and lower dosage LNG would result in uniform severe oligoazoospermia or azoospermia in normal men but would cause fewer adverse metabolic side effects. We conducted a randomized, placebo-controlled, single blind trial comparing 6 months of T enanthate (100 mg IM, weekly) plus LNG, 125 microg by mouth, daily (LNG 125; n = 18) or LNG, 250 microg by mouth, daily (LNG 250; n = 18) and compared these regimens with our previous study of the same dosage of T enanthate combined with placebo LNG (LNG 0; n = 18) or with 500 mg of LNG (LNG= 500; n = 18). All three combination regimens of T enanthate and LNG suppressed spermatogenesis more rapidly and resulted in significantly more uniform severe oligoazoospermia (<1 million/mL) than the T-alone regimen. Severe oligoazoospermia was achieved in 89% of the LNG 125, 89% of the LNG 250, and 78% of the LNG 500 groups, respectively, versus 56% of the men in LNG 0 (P < 0.05 for the combination groups vs. LNG 0), but there were no significant differences between the combination regimens (P = NS). All four groups gained significant weight compared with their baselines, although the gain tended to be greater as the dosage of LNG increased (2.0+/-0.9, 2.9+/-1.1, 3.6+/-1.0, and 5.4+/-1.0 kg gained, compared with baseline in the LNG 0, 125, 250, and 500 groups respectively; P < 0.05 compared with baseline). Serum levels of HDL cholesterol decreased in all of the groups, but the effect was larger as the dosage of LNG increased (4+/-4% vs. 13+/-4%, 20+/-3%, and 22+/-4% decrease in HDL levels from baseline in the LNG 0, LNG 125, LNG 250, and LNG 500 groups respectively; P = 0.06 for LNG 125 compared with LNG 0, and P < 0.05 for LNG 250 and LNG 500 compared with LNG 0). We conclude that 1) the combination of physiologic exogenous T enanthate and LNG suppresses spermatogenesis more effectively than T enanthate alone and that 2) the combination regimen of T enanthate plus lower dosage LNG suppresses sperm production comparably to T enanthate plus higher dosage LNG, while causing less weight gain and HDL cholesterol suppression. A combination regimen of physiologic testosterone plus a low dosage of levonorgestrel offers great promise as a safe and effective male contraceptive regimen. PMID- 10386823 TI - Evidence that in a physiological setting Sertoli cell number is the major determinant of circulating concentrations of inhibin B in the adult male rhesus monkey (Macaca mulatta). AB - The relationship between changes in Sertoli cell number and function and changes in circulating inhibin B concentrations was investigated following unilateral orchidectomy (UO) in the adult rhesus monkey. As expected, the 50% loss in Sertoli cells resulting from UO on day 0 was associated with a rapid and corresponding decline in plasma concentrations of inhibin B. The decrease in inhibin B levels was sustained until the remaining testis was removed on day 44, at which time a compensatory 50% increase (P < 0.05) in the number of round spermatids was evident in the absence of a change in Sertoli cell number. Moreover, Sertoli cell number and inhibin B levels among individual monkeys were highly correlated (r2 = 0.65, P < 0.002). Round spermatid number and inhibin B, however, were poorly correlated (r2 = 0.37, P < 0.04). These findings indicate that, in a physiological setting where the negative feedback control system governing the adult primate testis is operational, Sertoli cell number, rather than function, is the primary determinant of circulating inhibin B levels. PMID- 10386822 TI - Cell- and region-specific localization of lysosomal and secretory proteins and endocytic receptors in epithelial cells of the cauda epididymidis and vas deferens of the adult rat. AB - The epithelial cells lining the cauda epididymidis and vas deferens are active in endocytosis and have an abundance of lysosomes and a well-characterized secretory apparatus. However, little is known about the nature of lysosomal proteins contained within lysosomes, the types of receptors on the cell surface, and the types of proteins secreted by these cells. In the present study, cathepsins A, D, B, and sulfated glycoprotein (SGP)-1, well-characterized lysosomal proteins, as well as SGP-2, a secretory protein and low-density lipoprotein receptor-related protein-2 (LRP-2), an endocytic receptor, were immunolocalized at the light microscopic level within epithelial cells of the cauda epididymidis and vas deferens. Principal cells showed numerous intensely reactive lysosomes for cathepsins A, D, and SGP-1 in all regions of the cauda and vas deferens and for cathepsin B only in the cauda epididymidis. Basal cells were intensely reactive for cathepsin A, unreactive for cathepsins D and B, and weakly reactive for SGP-1 in the cauda region. In the vas deferens, these cells were intensely reactive for cathepsin A and SGP-1 and unreactive for cathepsin B; in the case of cathepsin D, basal cells were weakly reactive in the proximal vas deferens but intensely reactive in the middle and distal vas deferens. Clear cells, present in the cauda region and proximal vas deferens, were intensely reactive for cathepsin A, weakly reactive for SGP-1, and unreactive for cathepsins D and B, while narrow cells found mainly in the proximal vas deferens were intensely reactive for cathepsins A, D, and SGP-1 and unreactive for cathepsin B. Thus, the expression of different lysosomal enzymes in the cauda epididymidis and vas deferens is not only cell- but also region-specific, suggesting differences in the type of substrates internalized by these cells. SGP-2, a secretory protein, showed a checkerboardlike staining pattern in the cytoplasm of principal cells of the cauda epididymidis, while the cytoplasm of all principal cells were intensely reactive in the vas deferens. This type of reaction, as well as staining of sperm, suggests that SGP-2 is secreted into the lumen, where it functions in relation to sperm. The endocytic receptor LRP-2 was noted only on the apical surface of principal cells of the cauda and vas deferens and in spherical structures indicative of endosomes suggestive of their role in the uptake of various ligands, including SGP-2, for which it has a high binding affinity. Thus SGP-2 in the cauda and vas deferens is not only secreted but endocytosed by principal cells, suggestive of an active turnover in the lumen. In summary, the epithelial cells of the cauda and vas deferens show marked differences in expression of lysosomal proteins, SGP-2, and LRP-2 suggestive of differences in their functional activity while sperm are stored and protected in these regions. PMID- 10386824 TI - Pfiesteria toxin and learning performance. AB - Pfiesteria piscicida is an estuarine dinoflagellate involved with fish kills along the east coast of the United States. We previously documented a radial-arm maze learning deficit in rats exposed to Pfiesteria that may be related to cognitive deficits seen in humans after accidental Pfiesteria exposure. The current study elucidated important behavioral parameters of this deficit. There were six dose groups. Forty (10/group) adult female Sprague-Dawley rats were injected (s.c.) with a single dose of Pfiesteria taken from aquarium-cultured Pfiesteria (35,600, 106,800, or 320,400 Pfiesteria cells/kg of rat body weight or a cell-free filtrate of the 106,800 cells/kg dose). One control group (N = 10) was injected with saline and one (N = 10) with aquarium water not containing Pfiesteria. Half of the rats in each group were tested on an 8-arm radial maze in a standard test room, and the other half were tested on the radial maze in a sound-attenuating chamber. In the standard maze room, there was a significant effect of Pfiesteria (p < 0.05) impairing choice accuracy improvement over the first six sessions of training among rats administered 106,800, 320,400, and the 106,800 cells/kg filtered sample. In contrast, there was no indication of an effect of Pfiesteria when the rats were tested on the same configuration radial maze in the sound-attenuating chamber. After 18 sessions of training in one room, the rats were switched for six sessions of testing in the other room and finally were switched back to their original room for three sessions. There was a significant Pfiesteria-induced deficit when the rats were tested in the standard test room but not when they were tested in the sound-attenuating chamber. When the Pfiesteria-exposed rats were initially switched from the sound-attenuating chamber to the standard test room they performed significantly worse than controls, whereas Pfiesteria-treated rats switched from the standard test room to the sound-attenuating chamber did not perform differently from controls. These results suggest that the Pfiesteria-induced learning impairment may result from the negative impact of distracting stimuli. At the time of the learning impairment, no overt Pfiesteria-related effects were seen using a functional observational battery and no overall response latency effects were seen, indicating that the Pfiesteria-induced choice accuracy deficit was not due to generalized debilitation. In the initial use of the figure-8 maze in this line of research, the rats in the same Pfiesteria treatment groups that showed significant deficits in the radial-arm maze showed greater declines in activity rates in a 1-h figure-8 locomotor activity test. Both the 106,800 and 320,400 Pfiesteria cells/kg groups showed significantly greater linear trends of activity decline relative to tank water-treated controls. This reflected an initial slight hyperactivity in the Pfiesteria-treated animals followed by a decrease to control levels. Pfiesteria effects in the figure-8 maze and in early radial-arm maze training may be useful in a rapid screen for identifying the critical toxin(s) of Pfiesteria in future studies. PMID- 10386825 TI - Operant test battery performance in children: correlation with IQ. AB - The relationship between intelligence and money-(nickel-)reinforced operant behaviors were compared in 115 six year old children. The Operant Test Battery (OTB) consists of tasks thought to engender responses dependent upon specific brain functions that include motivation, color and position discrimination, learning, short-term memory, and time estimation. OTB endpoints were compared with Full Scale, Verbal and Performance IQ scores. Highly significant correlations were noted between several OTB measures (e.g., color and position discrimination accuracy) and IQ scores, but not in others (e.g., motivation task response rate). The results demonstrate the relevance of these measures as metrics of important brain functions. Additionally, since laboratory animals can readily perform these same tasks, these kinds of behaviors in laboratory animals should be useful in studying the effects of neuroactive/neurotoxic compounds on aspects of cognitive function in animals and in predicting adverse effects of such agents on related brain functions in humans. PMID- 10386826 TI - Learning and memory in rats gestationally and lactationally exposed to 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD). AB - Recently we reported that in utero and lactational exposure to 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) or coplanar polychlorinated biphenyls (PCBs) resulted in a reduction of errors on a radial arm maze (RAM) working memory task. The effect was more pronounced in males than in females. In this study, we further investigated the effects of in utero and lactational exposure to TCDD on learning and memory by testing male and female TCDD-exposed rats on three different spatial learning and memory tasks: the RAM, the Morris water maze (MWM), and spatial discrimination-reversal learning (RL), as well as on a nonspatial learning task, visual RL. Time-mated Sprague-Dawley rats were gavaged with either TCDD (0.1 microg/kg/day) or corn oil vehicle on gestation days 10-16. Litters were culled to eight on day 2 and weaned on day 21. Beginning on day 80, one male and one female from each litter were tested on the same RAM working memory task used in the previous study. Again, the TCDD-exposed male rats displayed a pronounced decrease in errors relative to control males. Following the RAM testing, the same animals were tested on the MWM, but no differences between the exposed and control rats were observed. Another male and female from each litter were tested on spatial RL on a T-maze. There were no differences between the exposed and control rats on this task. Following spatial RL, the same rats were tested on visual RL on the same maze. The exposed animals did not differ from controls on original learning, but took more trials to reach criterion on the first and second reversals. These results demonstrate a reliable, but task-specific, facilitation of spatial learning and memory in male rats exposed to TCDD during gestation and lactation. In contrast, both male and female TCDD-exposed rats showed a deficit in learning on the visual RL task. This pattern is consistent with that seen in earlier monkey studies. Perinatally TCDD exposed monkeys were facilitated on certain spatial tasks, but impaired on visual RL tasks. PMID- 10386827 TI - Effects of prenatal exposure to toluene on postnatal development and behavior in rats. AB - Development and neurobehavioral effects of prenatal exposure to toluene (CAS 108 88-3) were studied after exposing pregnant rats (Mol:WIST) to 1800 ppm of the solvent for 6 h daily on days 7-20 of gestation. Body weights of exposed offspring were lower until day 10 after parturition. Neurobehavioral evaluation of the pups revealed no effects on motor function (rotarod), activity level (open field), acoustic startle, and prepulse inhibition. Measurements of hearing function using auditory brain stem response revealed small effects in male exposed offspring. Performance in a Morris water maze during initial learning gave some indications of impaired cognitive functions, which was confirmed during further testing, especially in reversal and new learning. Effects on cognitive functions seemed most marked in female offspring. PMID- 10386828 TI - Effect of chronic prenatal ethanol exposure on nitric oxide synthase I and III proteins in the hippocampus of the near-term fetal guinea pig. AB - Chronic prenatal ethanol exposure suppresses nitric oxide synthase (NOS) enzymatic activity, in the hippocampus of the near-term fetal guinea pig at gestational day (GD) 62. The objective of this study was to determine if this decrease in NOS activity is the result of decreased NOS I and NOS III protein expression. Pregnant guinea pigs received oral administration of 4 g ethanol/kg maternal body weight/day (n = 8), isocaloric-sucrose/pair feeding (n = 8), or water (n = 8) from GD 2 to GD 61. The NOS I and NOS III protein expression and localization in the hippocampus were determined using Western blot analysis and immunohistochemistry, respectively. The chronic ethanol regimen produced fetal body, brain, and hippocampal growth restriction compared with the isocaloric sucrose/pair fed and water groups but did not affect the expression or localization of NOS I and NOS III proteins in the hippocampus. The decrease in NOS enzymatic activity induced by chronic prenatal ethanol exposure may be the result of posttranslational modification of NOS I and/or NOS III protein in the hippocampus of the near-term fetal guinea pig. PMID- 10386829 TI - Neurodevelopment after in utero amiodarone exposure. AB - It is not known whether amiodarone is neurotoxic to the fetus, as it is to adults. We evaluated neurodevelopment of a historical cohort (N = 10) of children exposed transplacentally to amiodarone. Scores on standardized tests of cognitive and language skills were compared (by Wilcoxon signed rank test) between eight toddlers and matched controls. It was not possible to obtain controls for older amiodarone-exposed children (aged 9.7 and 12.0 years), whose test results were compared descriptively with normative data. There was no difference in IQ scores between amiodarone-exposed toddlers and controls. All had favorable temperaments. However, amiodarone-exposed toddlers showed expressive language skills that were relatively poorer than verbal skills, when compared with controls (p = 0.046). One amiodarone-exposed toddler exhibited global developmental delay. The older amiodarone-exposed children had well-developed social competence, favorable global IQ scores, but problems with reading comprehension, written language, and arithmetic. This picture is reminiscent of the Nonverbal Learning Disability Syndrome. There may be neurotoxicity associated with transplacental exposure to amiodarone. Follow-up is warranted, although most mothers were happy with the development of their children. PMID- 10386830 TI - Toluene ototoxicity in rats: assessment of the frequency of hearing deficit by electrocochleography. AB - To identify the frequency range most sensitive to toluene-induced auditory damage, the auditory function of adult Long-Evans rats exposed to 1750 ppm of toluene (6 h/day, 5 days/week, 4 weeks), was tested by recording auditory-evoked potentials directly from the round window of the cochlea. The present electrocochleographic findings do not support a specific mid- to high-frequency loss of auditory sensitivity. On the contrary, the electrophysiologic data, obtained for audiometric frequencies ranging from 2 to 32 kHz, showed a hearing deficit not only in the mid-frequency region (12-16 kHz), but also in the mid-low frequency region (3-4 kHz). Actually, the effect of toluene was independent of the frequency in our experimental conditions. Histological analysis was consistent with electrophysiologic data because a broad loss of outer hair cells occurred in both mid- and mid-apical coil of the organ of Corti. PMID- 10386831 TI - Perinatal delta9-tetrahydrocannabinol exposure augmented the magnitude of motor inhibition caused by GABA(B), but not GABA(A), receptor agonists in adult rats. AB - We have extensively reported that delta9-tetrahydrocannabinol (delta9-THC) exposure results in changes in the adult functionality of dopaminergic neurons, in particular, mesotelencephalic pathways, although some changes are evident only after pharmacological challenges. In the present study, we have examined whether similar changes might be observed in gamma-aminobutyric acid (GABA) activity, in particular, in those regions where cannabinoid receptors have been reported to be located in GABA-containing neurons. To this end, we first examined GABA content and glutamic acid decarboxylase (GAD) activity in several brain regions of adult male and female rats that had been perinatally exposed to delta9-THC or oil. Delta9-THC exposure did not modify either GAD activity or GABA content in the ventral-tegmental area, nucleus accumbens, substantia nigra, caudate-putamen, and globus pallidus, thus suggesting no changes in the basal presynaptic activity of GABA-containing neurons. Second, we tested the motor response in the open-field test of these animals after a single injection of muscimol, a GABA(A) receptor agonist, baclofen, a GABA(B) receptor agonist, or vehicle. We observed that the motor inhibition caused by baclofen, in terms of decreased ambulation and stereotypy and increased inactivity, was more marked in magnitude in delta9-THC exposed males and females. This was not observed for the GABA(A) receptor agonist, muscimol, indicating a receptor specificity. To extend this observation, we also examined whether the potential differences in the behavioral response found in the above experiment might be due to changes at the level of the efficiency of the activation of these receptors by measuring basal and baclofen stimulated [35S]-guanylyl-5'-O-(gamma-thio)-triphosphate ([35S]-GTPgammaS) binding in adult male and female rats that had been perinatally exposed to delta9 THC or oil. However, our results were negative, because perinatal delta9-THC exposure did not increase baclofen-stimulated [35S]-GTPgammaS binding in the areas studied; in particular, in the substantia nigra, an area of interest for the interactions GABA(B) receptor/cannabinoid receptor. Collectively, the present results indicate that although perinatal delta9-THC did not produce any changes in GABA content and GAD activity in limbic and motor areas in adulthood, it did increase the behavioral response to GABA(B) receptor agonists. However, this increase was not due to changes in GABA(B) receptor activation of signal transduction mechanisms, as revealed the analysis of the percentage of stimulation by baclofen of [35S]-GTPgammaS binding in the substantia nigra and other structures of males and females perinatally exposed to delta9-THC. PMID- 10386832 TI - Prenatal binge-like alcohol exposure in the rat results in region-specific deficits in brain growth. AB - Children of women who abuse alcohol during pregnancy may be affected by varying degrees of neurological abnormality, even if they are not diagnosed with Fetal Alcohol Syndrome. The extent of the behavioral deficits of the affected offspring may be a function of several factors, such as the differential vulnerability of the various regions of the brain-to-alcohol insult. In this study, groups of timed-pregnant rats were exposed to different doses of alcohol (EtOH 2.25, EtOH 4.5, EtOH 6.5 g/kg/day) or control conditions (maltose dextrin solution or no treatment) from embryonic day 1 (E1: sperm positive) to E20. On E33 (usually postnatal day 10), all pups were perfused. Their brains were removed, dissected into forebrain, cerebellum, and brainstem, and weighed. Blood alcohol concentrations (BACs) were measured on 4 different days of gestation, but the peak BACs across gestation for the three alcohol-treated groups averaged 142, 294, and 413 mg/dl for the EtOH 2.25, EtOH 4.5, and EtOH 6.5 g/kg groups, respectively. Analysis of the body weight data indicated that pups in the EtOH 6.5 g/kg group had a greater somatic growth deficit than pups from all other groups. Although the whole brain, forebrain, cerebellum, and brainstem weights of pups in the EtOH 6.5 g/kg group were significantly smaller than those in the control groups, within-treatment group analyses indicated that the cerebella of pups in the EtOH 6.5 g/kg group were more severely affected than were their forebrains or brainstems. The analyses of the brain region to body weight ratios revealed again that the cerebellum-to-body-weight ratio of pups in the EtOH 6.5 g/kg group was more severely affected than the forebrain or brainstem to body weight ratios. Collectively, these data lend support to the view that gross regions of the brain are differentially vulnerable to alcohol insult during the first two trimesters equivalent, and suggest that the cerebellum is vulnerable to injury from exposure to high BACs during a developmental period other than the third trimester equivalent. PMID- 10386833 TI - Effects of cypermethrin on the electroencephalographic activity of the rat: a model of chemically induced seizures. AB - Cypermethrin is a potent representative member of the type II pyrethroid insecticides. This pyrethroid is used worldwide and has become a part of our environment. Until the present study, little information about its toxic effects in the central nervous system (CNS) was available. The aim of this study was, then, to determine the effects of repeated exposure to cypermethrin by means of assessing the electroencephalographic (EEG) activity in the rat. Cypermethrin was administered daily in a 300 mg/kg i.p. dose, below the LD50 value. After daily administration, the EEG activity was recorded and evaluated for 30 min. Paroxysmal epileptic activity appeared after the first and second days of cypermethrin administration. Frequency and numbers of bursts of epileptic activity also increased throughout the days of exposure to cypermethrin. Some of the paroxysmal events were present with behavioral anomalies, such as generalized tonic-clonic seizures. These effects are important because they could be related to the incidence of epileptic activity in humans chronically exposed to cypermethrin. PMID- 10386834 TI - Low-frequency hearing loss following perinatal exposure to 3,3',4,4',5 pentachlorobiphenyl (PCB 126) in rats. AB - Previous research has demonstrated the sensitivity of the developing rat to the ototoxic effects of exposure to Aroclor 1254. In this study we assessed the effects of developmental exposure to an individual PCB congener (3,3',4,4',5 pentachlorobiphenyl; PCB 126) on auditory function. Nulliparous Long Evans rats received either 0, 0.25, or 1.0 microg/kg/day (5 days/week) for 35 days prior to breeding and throughout gestation and lactation. Auditory thresholds for 0.5-, 1 , 4-, 8-, 16-, 32-, and 40-kHz tones were assessed in offspring on postnatal days (PND) 76-90. Perinatal maternal PCB 126 exposure caused low-frequency hearing deficits. Elevated auditory thresholds occurred in the 1.0 microg/kg/day treated group for 0.5- and 1-kHz tones, whereas thresholds were not significantly affected at any higher frequencies. These results are important in that the data implicate, at least partially, the coplanar PCBs in the developmental ototoxicity induced by Aroclor 1254. PMID- 10386835 TI - A stereotaxic atlas of the forebrain of the bank vole (Clethrionomys glareolus). AB - In this article part of the forebrain of the bank vole (Clethrionomys glareolus) is presented in stereotaxic coordinates. The stereotaxic procedure was performed as follows. With the vole's head mounted in a stereotaxic adaptor, internal reference tracks were made with a 0.5-mm diameter microdialysis cannula and India ink, 2 mm in front and 2.6 mm behind the skull landmark bregma. Brains were fixed for 72 h in 4% commercial formaldehyde in sodiumcacodylate buffer containing 1% CaCl2. To determine shrinkage they were weighed before and after fixation. After embedding in paraffin they were sectioned at 25 microm and stained with Nissl. Photomicrographs were taken from the brain of one animal while its frontal (antero-posterior) coordinates of five neural structures were compared with those of 12 other voles. Variability was also checked in lateral and vertical directions at frontal level -1.0 mm (relative to bregma). The results show that the distance between the two skull landmarks bregma and lambda correlates significantly and negatively with the antero-posterior position of each of the brain areas. On the basis of these results an equation is proposed to improve accuracy in locating neural structures that deviate due to biological variability. PMID- 10386836 TI - Time-dependent GABA-ergic activity in olfactory bulb and hypothalamus of proestrous rats. AB - Exposure of proestrous rats in the afternoon (1700h) or night (0100h) to intact and castrated male pheromones induced changes in olfactory bulbs and hypothalamic glutamic acid decarboxylase activity. Compared to the values found in control rats, the enzyme activity in the main olfactory bulb measured in the afternoon did not discriminate between the odor of intact or castrated males in the first hour of exposure, whereas in the accessory olfactory bulb, the enzyme activity was increased specifically by castrated male odor. At night, main bulb-enzyme activity was reduced by the intact male pheromone 1 h later, whereas accessory bulb-enzyme activity was increased by castrated male odor. Preoptic area enzyme activity was only modified by castrated male odor in the afternoon, whereas intact male pheromone exposure for 2 h decreased enzyme activity specifically at night. In the medial basal hypothalamus, both olfactory stimuli decreased afternoon enzyme activity, and on the contrary no changes were seen by nighttime. It is conclude that, in proestrous rats, the activity changes of main bulb and preoptic area GABA-ergic system evoked by intact male pheromone may be related to a facilitatory neural mechanism involved in the rat sexual motivation during the night. PMID- 10386837 TI - Glial cells in degenerating and regenerating optic nerve of the adult rat. AB - The glial cell reaction both in degenerating and regenerating adult rat optic nerve was studied by immunohistochemistry and electron microscopy. Degeneration in the optic nerve was achieved by complete transection, and the retinal stump was then analyzed. The regeneration was observed by autotransplantation of a sciatic nerve segment to the transected retinal stump. In both cases, optic nerve axons were labeled anterogradely with rhodamine, followed by immunohistochemical staining. Glial fibrillary acidic protein-positive astrocytes covered the transected end of degenerating optic nerve, whereas in the regenerating optic nerve they enwrapped axonal bundles emerging from the optic nerve stump and migrated together into the transitional zone intervening between the retinal stump and graft. In electron microscopy, direct attachment of astrocyte and Schwann cell was found within the transitional zone, whereby these cells were holding axons between them. Decrease of 04 immunoreactivity, which labels oligodendrocytes, was apparent in the transected end of retinal stump during the regeneration. The ED1 -positivity, which labels microglia/macrophages, was found in cells accumulated in the transitional zone of degenerating optic nerve, whereas during regeneration, ED1-immunoreactive cells were also distributed in the retinal stump. These results suggest that astrocytes, usually considered to interfere with optic nerve regeneration, change their characteristics in the presence of peripheral nerve graft and guide the regenerating axons in cooperation with Schwann cells. The response of oligodendrocytes and microglia/macrophages may also be modulated by peripheral nerve. PMID- 10386838 TI - Molecular cloning, expression, and activity of zebrafish semaphorin Z1a. AB - Semaphorins/collapsins are a large family of secreted and cell surface molecules that are thought to guide growth cones to their targets. Although some members are clearly repulsive to specific growth cones in vitro, the in vivo role of many of these molecules in vertebrate embryos is still unclear. As a first step towards clarifying the in vivo role of semaphorins/collapsins, we analyzed semaZ1a in the simple and well-characterized zebrafish embryo. SemaZ1a is a secreted molecule that is highly homologous to Sema III/D/collapsin-1, and it can collapse chick dorsal root ganglion growth cones in vitro. It is expressed in highly specific patterns within the developing embryo, which suggests that it influences outgrowth by a variety of growth cones including those of the posterior lateral line ganglion. Consistent with this hypothesis, the peripherally extending growth cones of posterior lateral line neurons retract and partially collapse during normal outgrowth. PMID- 10386839 TI - Individual differences in behavioral reactivity: correlation with stress-induced norepinephrine efflux in the hippocampus of Sprague-Dawley rats. AB - The present studies investigate the hypothesis that the locus coeruleus norepinephrine (LC-NE) system plays a role in the neural substrates underlying individual differences in behavioral reactivity to stress. Individuals were selected from a random sample of Sprague-Dawley rats and categorized as a high responder (HR), middle responder (MR), or low responder (LR) based on the initial locomotor response to a novel open field. Rats with behavioral scores at least 1 SD away from the mean for the subject sample were categorized as HR or LR rats. Middle responder rats exhibited locomotor scores representative of the mean locomotor activity of the population sample. Locomotor activity scores measured 6 days after the initial determination were similar to scores obtained in the original screening, suggesting that the locomotor response to novelty is a stable individual trait. Additionally, locomotor activity during the dark phase of the diurnal cycle was not different among the groups, suggesting that differences in locomotor activity in response to a novel open field are an index of behavioral reactivity to the stressful situation rather than an indicator of global differences in motoric activity. In vivo microdialysis was used to measure extracellular levels of hippocampal NE in the hippocampus. During baseline conditions, the efflux of hippocampal NE was similar among HR, MR, and LR rats. In response to tail-pinch stress, hippocampal NE release was elevated in all groups. This response was significantly greater in HR compared to LR rats. Across all groups, locomotor response in the novel open field was significantly correlated with the magnitude of NE release in response to subsequent application of tail-pinch stress. In contrast, administration of 1.5 mg/kg, i.p., amphetamine resulted in a similar elevation of extracellular NE level among HR, LR, and MR rats. These data suggest that activation of the LC-NE system may be involved in determining the behavioral response of individuals to environmental stress. PMID- 10386840 TI - Changes in hippocampal theta following intrahippocampal corticotropin-releasing hormone (CRH) infusions in the rat. AB - Hippocampal theta activity is a large amplitude, sinusoidal wave that occurs during attentive immobility and exploratory behaviour in the rat, and it is thought to be involved in memory formation. Recent reports suggest that corticotropin-releasing hormone (CRH) has pro-mnemonic effects in rodents. Because memory-enhancing substances/manipulations generally alter either theta frequencies or amplitudes, these variables were monitored in urethane anaesthetised rats following intrahippocampal infusions of CRH. Adult male, Lister hooded rats were implanted with a hippocampal recording electrode and a guide cannula, both aimed at the dentate gyrus. When CRH was infused into the hippocampus, the main change in the hippocampal EEG was a slow onset increase in the amplitude of spontaneous theta and, paradoxically, a significant decrease in the amount of time spent displaying theta. These data suggest that CRH has the ability to modulate ongoing hippocampal theta, but, considering the slow effect, the involvement of hippocampal CRH receptors is suspect. Regardless of locus, the described electrophysiological changes suggest that hippocampal cholinergic systems may play a role in the memory-enhancing effects of CRH. PMID- 10386841 TI - The effect of morphine on responses of nucleus ventroposterolateralis neurons to colorectal distension in the rat. AB - In 71 halothane-anesthetized rats, we characterized the responses of single neurons in the nucleus ventroposterolateralis (VPL) of the thalamus to a noxious visceral stimulus (colorectal balloon distension; CRD) and studied the effects of intravenous morphine on these responses using standard extracellular microelectrode recording techniques. One hundred nine neurons were isolated on the basis of spontaneous activity. Sixty-four (59%) responded to CRD, of which 52 (81 %) had excitatory and 12 (19%) had inhibitory responses. Neurons showed graded responses to graded CRD pressures (20-100 mmHg), with maximum excitation or inhibition occurring at 80 mmHg. Responses to noxious (pinch, heat) and innocuous (brush, tap) cutaneous stimuli were studied in 95 of the VPL neurons isolated. Eighty-three of these neurons (48 CRD responsive and 35 CRD nonresponsive) (87%) had cutaneous receptive fields, of which 96% were small and contralateral and 4% were large and contralateral or bilateral. Ninety-four percent of these neurons responded to both noxious and innocuous cutaneous stimulation, and 6% responded to only noxious stimulation. No neurons responded solely to innocuous stimulation. Cumulative doses of morphine (0.125, 0.25, 0.5, 1, and 2 mg/kg, i.v) produced statistically significant dose-dependent attenuation of neuronal responses to CRD. Naloxone (0.4 mg/ kg, i.v.) reversed the effects of morphine. Morphine and naloxone had no significant effects on spontaneous activity. These data support the involvement of VPL neurons in visceral nociception and are consistent with a role of VPL in sensory discriminative aspects of nociception. PMID- 10386842 TI - The developmental changes of the "paraclaustral reservoir" of migrating cells in the rat brain: a study using morphometric and in situ DNA end labeling techniques. AB - A distinct group of small cells lying in the ventral part of the external capsule in the rat brain is clearly visible at birth. On the basis of its location (medially to the prepiriform claustrum) and probably its function (as a source of neurons for adjacent structures), we define this nucleus as the "paraclaustral reservoir". The present study reveals the cellular changes of the paraclaustral reservoir during postnatal development of the rat brain using unbiased morphometry and in situ DNA end labeling. During the first 4 days after birth the density and total number of cells in the paraclaustral reservoir were stable; after this period a decrease of these parameters was observed until the complete disappearance of this structure at the end of first postnatal week. The rather low number of TUNEL (TdT mediated dUTP nick end labeling of fragmented DNA) positive nuclei in the paraclaustral reservoir suggests that apoptosis is not a crucial mechanism leading to decay of this structure. PMID- 10386843 TI - Modulation of synaptic transmission by nicotine and nicotinic antagonists in hippocampus. AB - Using rat hippocampal slices, we studied the effects of nicotine and three antagonists of neuronal nicotinic receptors on excitatory and inhibitory transmission. We report that nicotine at concentrations between 0.5 and 100 microM enhanced excitatory synaptic responses and increased the size of the presynaptic fiber volley. This effect was reproduced by three neuronal nicotinic receptor antagonists: dihydro-beta-erythroidine, methyllycaconitine and mecamylamine. In contrast, nicotine, but not nicotinic antagonists, produced a dual effect on inhibition: nicotine enhanced gamma-aminobutyric-acid A (GABA(A)) receptor-mediated synaptic responses at low concentration (0.5 microM) and blocked them at high concentration (100 microM). We conclude that the excitatory effects of nicotine are reproduced by nicotinic receptor antagonists, thereby suggesting that these effects might be mediated through receptor desensitization. These results also indicate that nicotine differentially affects GABAergic inhibition at low and high concentrations-effects that are not reproduced by antagonists. PMID- 10386844 TI - Chemical stimulation of the laryngopharynx increases Fos-like immunoreactivity in the rat hypothalamus and amygdala. AB - Using immunohistochemical detection of the Fos protein as a cellular marker of neuronal activation, we examined forebrain areas that may be activated upon chemical stimulation of the laryngeal opening. Anesthetized rats were subject to multiple infusions of a chemical solution into the laryngopharynx. These animals were compared to two control groups: a surgical control group in which the animals were subject to the surgical procedure but received no stimulus infusions and a flow control group in which physiological saline replaced the chemical stimulus. Comparing the numbers of Fos-like-immunoreactive neurons in regions of the forebrain across groups revealed that infusing the chemical stimulus solution into the laryngopharyngeal opening selectively increased the number of Fos-like immunoreactive nuclei in the paraventricular nucleus of the hypothalamus and the central nucleus of the amygdala, two autonomic-visceral related forebrain regions. Within the paraventricular nucleus of the hypothalamus, Fos-like immunoreactive nuclei were significantly increased in the parvocellular subdivision while in the central nucleus of the amygdala, significant increases in Fos-like-immunoreactive nuclei were limited to the lateral capsular subdivision. These data suggest that in the rat laryngopharyngeal chemosensory stimulation activates forebrain regions that receive oral sensory information and are involved in visceral and autonomic functions. PMID- 10386845 TI - The effects of ibogaine on dopamine and serotonin transport in rat brain synaptosomes. AB - Ibogaine has been shown to affect biogenic amine levels in selected brain regions. Because of the involvement of these neurotransmitters in drug addiction, the effects of ibogaine on biogenic amine transport may contribute to the potential anti-addictive properties of ibogaine in vivo. With rat brain synaptosomes as our experimental system, we measured the effects of ibogaine on the uptake and release of dopamine (DA) and serotonin (5-HT). Ibogaine competitively blocked both DA and 5-HT uptake with IC50 values of 20 microM at 75 nM 3H-DA and 2.6 microM at 10 nM 3H-5-HT. Ibogaine had no effect on K+-induced release of 3H-DA from preloaded synaptosomes, but 20 microM and 50 microM ibogaine inhibited roughly 40% and 60%, respectively, of the K(+)-induced release of 3H-5-HT from preloaded synaptosomes. In the absence of a depolarizing stimulus, ibogaine evoked a small release of 3H-DA but not 3H-5-HT. These relatively low-potency effects of ibogaine on DA and 5-HT uptake in synaptosomes are consistent with the low binding affinity of ibogaine that has been previously reported for DA and 5-HT transporters. Our results show that if ibogaine modulates DA and 5-HT levels in the brain by directly blocking their uptake, then a concentration of ibogaine in the micromolar range is required. Furthermore, if the anti-addictive effects of ibogaine require this concentration, then ibogaine likely exerts these effects through a combination of neurotransmitter pathways, because binding affinities and functional potencies of ibogaine in the micromolar range have been reported for a variety of neuronal receptors and transporters. PMID- 10386846 TI - Tolerable hearing aid delays. I. Estimation of limits imposed by the auditory path alone using simulated hearing losses. AB - OBJECTIVE: When people who wear hearing aids speak, there are three paths by which they hear their own voices: 1) through the air and leakage around the earmold; 2) via the solid structures of their head; 3) through the air to the hearing aid microphone, and then through the aid circuitry. These paths involve different time delays. Digital processing introduces delays in path 3 from a few to several tens of milliseconds, which could lead to a range of disturbing effects. We examined one purely auditory effect, namely hearing speech through all three of these paths. Subjective disturbance was measured as a function of delay in path 3 using simulations of hearing loss and a simulated hearing aid. With increasing hearing loss, the loudness of sound heard via paths 1 and 2 decreases, and the aid user relies more on path 3. The disturbance produced by the delay then might be less perceptible. To test this idea, four different hearing losses were simulated, varying from mild to moderately severe. DESIGN: Each of two talkers was fitted with a closed earmold, and simultaneous above-ear and in-ear recordings were made of each talker reading prose. The above-ear signal was amplified using a simulated hearing aid with 4-channel full dynamic range compression; compression ratios and gains were selected using an algorithm based on the absolute thresholds used in the simulations of hearing loss. The resultant output was then mixed with the in-ear signal with one of five values of delay, and the combined signal was processed using the four simulations of hearing loss. The resulting stimuli simulated for normal-hearing listeners the experience of having a hearing impairment and listening through a hearing aid while talking, except that the talker's voice was not that of the listener. Twenty normally hearing subjects gave subjective ratings of the disturbance of the echo for each delay and each simulated hearing loss. RESULTS: Disturbance ratings generally increased monotonically with increasing delay. Average results show that delays are rated as "disturbing" for values between 20 and 30 msec for mild to moderate losses. For a moderately severe loss, the rating "disturbing" was not quite achieved at 40 msec. For moderate losses, a speaker with low fundamental frequency (f0)(70 to 140 Hz) was less disturbing than a speaker with a medium f0, (100 to 180 Hz). This effect reversed for the mildest loss for low values of delay. CONCLUSIONS: The auditory effects of delays between bone conducted sound and aid-conducted sound are likely to become disturbing for delays exceeding 20 msec. Somewhat longer delays may be tolerable for moderate to severe hearing losses. These delays are smaller than the delays at which audio visual integration is disrupted. PMID- 10386847 TI - Cross-modality matching: a tool for measuring loudness in sensorineural impairment. AB - OBJECTIVE: The main goal of this study was to establish the viability of cross modality matching (CMM) for the measurement of individual loudness functions in sensorineural-impaired hearing. To achieve this goal, CMM was tested rigorously to assess four measurement requirements: 1) internal consistency; 2) small relative variance across listeners; 3) test-retest reliability; and 4) data validity. DESIGN: The measurements involved two sensory continua: perceived length and loudness. Sensation-magnitude functions were generated for all listeners from absolute magnitude estimation (AME) of perceived length, from CMM between loudness and perceived length, and from AME and absolute magnitude production (AMP) of loudness. A total of 211 listeners, 83 with normal hearing at the stimulus frequency and 128 with a diagnosis of cochlear impairment of long duration, performed all four magnitude-scaling tasks. Supplementary loudness matches also were obtained. RESULTS: Based on the analysis of data, the following results were obtained. First, in accord with loudness measures in normal hearing, loudness measures in cochlear-impaired hearing showed that individuals with bilateral impairments can produce internally consistent loudness data. Second, over the stimulus range where cochlear impairment steepens the loudness function, in a log-log plot loudness slopes derived from CMM, like those obtained from AME and AMP of loudness, were larger in cochlear-impaired hearing than in normal hearing. However, the results of CMM were typically less variable than those obtained from AME and AMP of loudness, permitting a clear-cut distinction between loudness growth rates (slopes) in normal and cochlear-impaired hearing. Third, the results showed that within a cochlear-impaired population, much of the intersubject variability of the slope of the loudness function can be ascribed to the heterogeneity of individual thresholds. Consistent with loudness matching, the size of the slopes increased with the degree of hearing loss. The dependence of the size of the slopes on the degree of hearing loss was observed for hearing losses as large as 75 dB. Fourth, test-retest reliability data for 36 listeners showed that CMM can yield reliable and stable loudness-growth measures in cochlear-impaired hearing over the long term. Finally, equal-sensation matches obtained directly from loudness matching closely agreed with those obtained indirectly from magnitude scaling, indicating that CMM is a valid method for the measurement of loudness magnitudes. CONCLUSIONS: Taken together, the results demonstrate that CMM can yield stable, accurate, and robust loudness growth measures in cochlear-impaired hearing. Given its apparent reliability, validity, and ease of application, CMM has the potential to become a powerful tool for assessing the growth of loudness in a clinical population. Loudness-level functions derived from CMM may well be important for determining the frequency gain response of a hearing aid that most closely compensates for the distorted input-output function of the impaired auditory system. PMID- 10386848 TI - Aided growth of masking for speech and nonspeech signals. AB - OBJECTIVE: Growth of masking (GOM) functions for speech and nonspeech signals were obtained in normal and impaired ears, with an emphasis on aided listening conditions. The purpose of the investigation was to determine whether electroacoustic contrasts between hearing aid circuits would manifest themselves in aided GOM functions and to determine whether amplification alters the slopes of GOM functions in impaired ears. DESIGN: Five normal-hearing and 10 hearing impaired listeners with sloping high-frequency losses participated. GOM functions were obtained in the sound field using a two-interval forced choice forward masking paradigm. Four sets of test signals consisted of narrowband noise maskers and sinusoidal probes. Two additional stimulus sets consisted of phonemes used as masker and probe. Both on-frequency and off-frequency masking conditions were examined. Test circuits included wide dynamic range compression (low compression threshold and low compression ratio) and output compression limiting (high compression threshold and high compression ratio), each adjusted to the DSL[i/o] prescriptive target. GOM slopes were calculated for normal-hearing and hearing impaired subjects unaided, and for impaired subjects aided, in an effort to determined whether amplification compensates for abnormal growth of masking and whether electroacoustic contrasts between circuits differentially affect level dependent masker effectiveness. RESULTS: Results indicated large differences between normal and impaired functions unaided. Under aided conditions, narrowband noise masked thresholds were reduced re: unaided at lower masker levels but not at higher masker levels. Masked thresholds for phoneme maskers were reduced at all masker levels. In general, aided GOM slopes changed in the direction of normal but did not match normal slopes. Aided results were highly dependent on stimulus test conditions and were consistent with the compression thresholds and release times of the aids under test. Circuit contrasts were more apparent when phonemes were used as masker and probe than when conventional signals were used. CONCLUSION: Results suggest that GOM functions can be altered by amplification in ways that are consistent with the electroacoustic characteristics of the circuits being used. Although these data address only a small number of conditions, they suggest a possible application for aided growth of masking in the evaluation of hearing aid performance. PMID- 10386849 TI - Within-patient longitudinal speech reception measures with continuous interleaved sampling processors for ineraid implanted subjects. AB - OBJECTIVE: To assess within-subject changes in speech reception over time in a group of Ineraid subjects fitted with continuous interleaved sampling (CIS) wearable processors fabricated in Geneva. To compare asymptotic performance between CIS and Ineraid processors for the same subjects. DESIGN: Twelve patients, all users of the 4-channel Ineraid cochlear implant system for several years and with no previous experience of CIS processors in daily life, were equipped with Geneva Wearable Processors programmed to implement a high-rate CIS sound processing strategy using four to six channels. Their speech reception performance with CIS processors was monitored over a period of 1 yr with consonant and vowel identification tests. For comparison, speech reception performance also was measured with Ineraid processors before switching to CIS and after 6 mo of non-use of Ineraid processors. RESULTS: At fitting, CIS processors produced significantly better consonant identification but no better vowel identification. Subsequently, consonant and vowel scores with CIS processors improved progressively to asymptote after 6 mo of daily use. At 6 mo and beyond, performance with CIS processors was significantly superior to that obtained with Ineraid processors on both consonant and vowel identification tests. Control tests made with Ineraid processors after 6 mo of non-use of the device yielded results that were indistinguishable from those obtained before the study. CONCLUSIONS: The full potential of the CIS strategy is not revealed at fitting. Accumulation of daily experience provides significant improvements, asymptotic performance being reached after about 4 to 6 mo of use. All Ineraid users might greatly benefit from CIS processors. PMID- 10386850 TI - Clinical findings for a group of infants and young children with auditory neuropathy. AB - OBJECTIVE: To examine the prevalence of auditory neuropathy in a group of infants at risk for hearing impairment and to present an overview of the clinical findings for affected children. DESIGN: Results for 20 subjects who showed repeatable cochlear microphonic potentials in the absence of click-evoked auditory brain stem responses are included in this study. Behavioral and steady state evoked potential thresholds were established in each case. Where possible, otoacoustic emission and speech perception results (unaided and aided) also were obtained. RESULTS: One in 433 (0.23%) of the children in our series had evidence of auditory neuropathy. The audiometric findings for these subjects varied significantly, with behavioral thresholds ranging from normal to profound levels. Discrimination skills were also variable. Approximately half of the subjects showed little understanding, or even awareness, of speech inputs in both the unaided and aided conditions. There were, however, a number of children who could score at significant levels on speech discrimination tasks and who benefited from the provision of amplification. CONCLUSION: The results suggest that auditory neuropathy is more common in the infant population than previously suspected. The effects of neuropathy on auditory function appear to be idiosyncratic, producing significant variations in both the detection and discrimination of auditory signals. As such, the management of children with this disorder must allow for individual differences. PMID- 10386851 TI - Median averaging of auditory brain stem responses. AB - OBJECTIVE: The primary aim of this study is to demonstrate the feasibility of acquiring auditory evoked potentials (AEPs) by median averaging and study its performance under various recording conditions. The auditory brain stem response (ABR) was used as the AEP of choice because it has the poorest signal to noise ratio (SNR) with inherent high susceptibility to extraneous noise. Secondary aim is to evaluate the characteristics of the median ABRs in comparison with the conventional mean averaged ABRs. DESIGN: Single sweep responses to clicks obtained from four subjects at 5 dB steps were saved in hard disk and used for off-line mean and median averaging. The characteristics of the median averaging technique were investigated by manipulating the averaging procedure using the same set of single sweep recordings and comparing them with the mean averaged responses. The effects of analog to digital input resolution (bit size) was simulated computationally by increasing quantization. RESULTS: The results showed that AEPs with low SNRs such as the ABR can be successfully acquired using median averaging with about the same number of sweeps as was required for mean averaging, provided the EEG signal is digitized with a high number of bits. The resulting waveform generally contained more identifiable waves than the corresponding mean average and had a high-frequency noise superimposed on it. This high-frequency noise was successfully filtered out using a digital, running mean smoothing filter. The median average showed an advantage over the mean average when occasional artifacts were recorded. CONCLUSION: The results showed that ABRs can be acquired successfully by median averaging provided EEG is digitized with high bit size. Compared with conventional mean averaging, median averaging is less sensitive to infrequent, externally and internally generated noise that plagues conventional techniques and may help improve wave identification. PMID- 10386852 TI - Automatic detection of frequency changes depends on auditory stimulus intensity. AB - OBJECTIVE: A cortical cognitive auditory evoked potential, mismatch negativity (MMN), reflects automatic discrimination and echoic memory functions of the auditory system. For this study, we examined whether this potential is dependent on the stimulus intensity. DESIGN: The MMN potentials were recorded from 10 subjects with normal hearing using a sine tone of 1000 Hz as the standard stimulus and a sine tone of 1141 Hz as the deviant stimulus, with probabilities of 90% and 10%, respectively. The intensities were 40, 50, 60, 70, and 80 dB HL for both standard and deviant stimuli in separate blocks. RESULTS: Stimulus intensity had a statistically significant effect on the mean amplitude, rise time parameter, and onset latency of the MMN. CONCLUSION: Automatic auditory discrimination seems to be dependent on the sound pressure level of the stimuli. PMID- 10386853 TI - Neuroaudiological effects in a case of fatal dimethylmercury poisoning. AB - OBJECTIVE: The audiological examination of this patient was to determine the degree and type of hearing loss that was incurred from apparent dimethylmercury poisoning. DESIGN: This was a single subject case study design. Audiologic tests were selected to help determine sensory from neural and/or central auditory system dysfunction. RESULTS: This patient demonstrated an inability to understand speech, both in formal and informal assessments, yet relatively good hearing sensitivity for pure tones bilaterally. Distortion product otoacoustic emissions showed only minimal deficits in each ear. The auditory brain stem response was abnormal bilaterally, indicating neural and/or central involvement. CONCLUSION: Dimethylmercury poisoning, in this case, resulted in compromise of the auditory neural system with little effect on the sensory (cochlea) mechanism. PMID- 10386854 TI - Regulation of interleukin-8 gene expression. AB - Interleukin-8 (IL-8), a member of the CXC chemokine family, is an important activator and chemoattractant for neutrophils and has been implicated in a variety of inflammatory diseases. IL-8 is secreted in a stimulus-specific manner by a wide variety of cell types and is regulated primarily at the level of gene transcription. Functional studies indicate that IL-8 transcriptional responses to proinflammatory mediators are rapid and require only 100 nucleotides of 5' flanking DNA upstream of the TATA box. Within the IL-8 promoter sequence are DNA binding sites for the inducible transcription factors AP-1, NF-IL-6, and NF kappaB. Transcription factors in these families bind the IL-8 promoter as dimers, and several distinct subunit combinations have been identified as important for IL-8 transcription. In addition, these factors can act in concert to synergistically activate the IL-8 promoter. AP-1 and NF-IL-6 physically interact with NF-kappaB, and functional cooperativity among the factors appears to be critical for optimal IL-8 promoter activity in different cell types. IL-8 transcription appears to be activated by a promoter recruitment mechanism where inducible transcription factor binding to the IL-8 promoter is required for binding of constitutively active TATA box-binding proteins and formation of a stable preinitiation complex. This review discusses the regulatory role these higher-order synergistic interactions play in IL-8 transcription and in generation of the stimulus-specific and cell type-specific patterns of IL-8 expression. PMID- 10386855 TI - A novel mechanism for cytokine regulation: screening, selection, and characterization of anticytokine monoclonal and polyclonal autoantibodies. AB - We have recently described an immunoregulatory mechanism involving release of neutralizing autoantibodies (Aab) to cytokines during bacterial infections. Intraperitoneal inoculation of Haemophilus influenzae type b (Hib) into Sprague Dawley rats resulted in high levels of inflammatory mediators early after infection. Increased titers of cytokine Aab were observed, with a peak at day 7. We cloned Aab-producing B cells. Screening of the clones with five different cytokines resulted in detection of Aab-producing clones reactive with each cytokine. After repeated subcloning, monoclonal Aab (mAab) were selected and characterized for their specificity, isotypes, and affinities. To elucidate regulatory importance, mAab to interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) dose-dependently inhibited IFN-gamma-induced MHC expression by peritoneal macrophages and TNF-alpha-induced thymocyte proliferation, respectively. Fab fragments exhibited binding and neutralizing effects, confirming specificities. Cross-reactivity with other rat cytokines was excluded. Pools of clones containing several mAab to each cytokine were obtained and served as polyclonal Aab. The relative affinity of the Aab was determined and found to be of high index. The characterized Aab were tested in methodologic assays for cytokine detection, revealing that some Aab were useful in a cell release capturing (CRC) ELISA. PMID- 10386856 TI - Interferon enhances the activity of the anticancer ribonuclease, onconase. AB - Interferons (IFN) are biologic agents involved in the antiviral response and the inhibition of tumor growth. Biochemical pathways of IFN action include the double stranded RNA-activated oligoadenylate synthetase, RNase L, and double-stranded RNA-dependent protein kinase (PKR). Extracellular ribonucleases, especially onconase, also display antiviral and antitumor properties and involve degradation of RNA. We find that IFN increases the anticancer activity of onconase. These two agents work synergistically, and the effect is seen at the level of translation probably because of the degradation of tRNA. PMID- 10386857 TI - Induction of mucosal immune responses by administration of liposome-antigen formulations and interleukin-12. AB - We examined the effect of interleukin-12 (IL-12) on the induction of mucosal immune responses following intranasal immunization with liposome-antigen formulations. We assessed the immune response to two recombinant glycoproteins (gD and gB) from bovine herpesvirus type 1 (BHV-1). Positively charged liposomes induced significantly higher gD-specific IgA titers than did immunization with antigen alone. This liposome formulation was selected to further assess the ability of IL-12 to influence mucosal immune responses. Intranasal immunization with IL-12 gD-liposome formulations did not alter the induction of mucosal immune responses. However, a significant increase in anti-gD antibody responses was induced in serum after intranasal immunization with IL-12 gD-liposome when compared with animals immunized with gD-liposomes. Mucosal antibody responses induced by a subcutaneous priming followed by an intranasal boost were significantly higher than those induced by two intranasal immunizations with the same IL-12 liposome-gD formulations. Furthermore, this immunization protocol resulted in the induction of high levels of interferon-gamma (IFN-gamma) in the lungs of subcutaneously primed mice. These findings indicate that the immunomodulatory effects of IL-12 influenced immune responses to a vaccine antigen when delivered intranasally and that these responses can be further enhanced by subcutaneous priming. PMID- 10386858 TI - Failure to detect antiviral activity in serum and plasma of healthy individuals displaying high activity in ELISA for IFN-alpha and IFN-beta. AB - The presence of constitutively produced interferon (IFN)-alpha in the blood of healthy individuals has been the subject of contradictory discussions for years. Immunologic as well as biologic test procedures have demonstrated striking differences regarding serum IFN-alpha under physiologic conditions. We investigated the presence of immunoreactive IFN-alpha in serum samples of 923 healthy blood donors by means of a widely used commercially available ELISA. Of these, 254 (27.5%) exhibited detectable serum IFN-alpha levels. The sera of 85.1% of these people also contained IFN-beta. Both IFN were also demonstrated in EDTA anticoagulated plasma. However, none of these samples exhibited any antiviral effect on human A549 lung carcinoma cells challenged with encephalomyocarditis virus. Samples with high IFN-alpha ELISA activity did not abolish the antiviral action of added natural IFN-alpha, thus excluding IFN-alpha inhibitory factors. The experiments suggest that the detected compounds probably did not represent IFN-alpha but were the result of a cross-reaction with unknown serum components. A variety of disorders has been associated with elevated serum IFN-alpha levels that in most cases were detected by ELISA. In view of our data, these findings need to be carefully reevaluated. For the purpose of monitoring IFN-alpha levels in therapy of atopic, autoimmune, or malignant disorders, an appropriate detection system for IFN-alpha is advisable. PMID- 10386859 TI - IFN-beta interferes with the differentiation of dendritic cells from peripheral blood mononuclear cells: selective inhibition of CD40-dependent interleukin-12 secretion. AB - We studied the effects of interferon-beta (IFN-beta) on the differentiation of dendritic cells (DC) obtained by culturing plastic-adherent peripheral blood mononuclear cells (PBMC) from a total of 30 healthy volunteers in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL 4). First, we found that the addition of IFN-beta at the initiation of the culture did not modify DC morphology but caused a reproducible and statistically significant upregulation of HLA-DR, CD86, and CD80 surface expression. CD1a expression was significantly reduced, and CD40 expression was unchanged. We then determined the influence of IFN-beta on the production of cytokines by DC. DC differentiated in the presence of IFN-beta secreted significantly less IL-12 (p40 and p70) both spontaneously and on activation by fibroblasts transfected with the CD40L gene. This effect of IFN-beta was dose dependent and selective, as it was not observed for IL-6, IL-8, and tumor necrosis factor-alpha (TNF-alpha). As a consequence, DC differentiated in the presence of IFN-beta induced significantly less IFN-gamma secretion by alloreactive T cells, whereas they were more efficient than control DC in eliciting IL-5 secretion. We conclude that the direct action of IFN-beta on DC causes inhibition of their ability to secrete IL 12 in response to CD40 ligation and to elicit Th1 type responses. PMID- 10386860 TI - Augmentation of verotoxin-induced cytotoxicity/apoptosis by interferon is repressed in cells persistently infected with mumps virus. AB - Verotoxin type 2 (VT2) produced by enterohemorrhagic Escherichia coli (EHEC) has been shown to have high cytotoxic potency toward several human B lymphoid cell lines with and without Epstein-Barr virus (EBV). Cell death, apoptosis induced by VT2, is closely correlated with the expression of receptor molecule Gb3/CD77, recognized by the toxin, but not with the infection or presence of EBV. Pretreatment of cells with interferon-alpha (IFN-alpha) for 24 h resulted in augmentation of apoptosis by VT2. Pretreatment within 8 h, however, was not effective. It has been reported that IFN-alpha-induced apoptosis is correlated with the induction of the 2',5'-OAS/RNase L system or dsRNA-activated protein kinase (PKR) or both. We have established persistent infection in both Akata and P3HR-1 cells with mumps virus. The persistently infected cell lines, P3HR-MP2 and Akata-MP2, showed poor induction of 2',5'-OAS and PKR in response to IFN-alpha. Augmentation of VT2-induced apoptosis by IFN-alpha was not found in the cell lines P3HR-MP2 and Akata-MP2. Therefore, these findings were interpreted to indicate that augmentation of VT2-induced apoptosis by IFN-alpha may be mediated by PKR and the 2',5'-OAS/RNaseL system. It is also suggested that mumps virus can suppress apoptosis and establish persistent infection. PMID- 10386861 TI - Genomic organization and chromosomal localization of a new member of the murine interferon-induced guanylate-binding protein family. AB - An RNA species has been identified whose nucleotide sequence is closely related to the mRNA encoding the murine interferon (IFN)-induced guanylate-binding protein-1 (mGBP1) and an mRNA encoding an isoprenylated protein that is constitutively expressed in various organs in the rat. Sequence analysis of the gene encoding this newly identified RNA reveals that in its 5'-region it is identical to a DNA fragment reported to represent the 5'-region of a gene termed mGBP2. In light of this homology, we term this newly identified gene product mGBP2. mGBP2 is inducible following IFN treatment in animals bearing Gbp1a alleles, in which mGBP1 is transcriptionally upregulated by IFN treatment, as well as in animals bearing Gbp1b alleles, in which mGBP1 is not induced in response to IFN treatment. The genomic organizations of the genes encoding mGBP1 and mGBP2 are similar, and the nucleotide sequences of their IFN-responsive-like elements and their relative locations are conserved. Gbp1 and Gbp2 map to a genetically indistinguishable site on the distal arm of chromosome 3. PMID- 10386862 TI - Extensive alternative splicing of interleukin-7 in malignant hematopoietic cells: implication of distinct isoforms in modulating IL-7 activity. AB - Interleukin-7 (IL-7) plays a pivotal role in early stages of normal B and T cell development. In addition, IL-7 stimulates the proliferation of both antitumor reactive cells and a number of T and B cell malignancies, underlining its significance for leukemogenesis. However, its exact role in the process of pathologic maturation of lymphocytes and regulation of the immune response is not completely understood. As alternative splicing of pre-mRNA has been shown to be involved in the control of gene expression, and splicing-derived protein isoforms with antagonistic activity have been found, we assessed the mRNA-expression of IL 7 and its previously described alternative splice variant lacking exon 4, IL 7delta4, in leukemic cells from children with acute lymphoblastic leukemia (ALL). PCR of full-length IL-7 cDNA enabling the competitive amplification of both variants led to the amplification of diverse unexpected PCR products. The sequence data demonstrated the existence of three additional in-frame splice variants resulting from exon skipping of exon 3 or exon 5 or both in combination with exon 4. We named these IL-7delta3/4, IL-7delta4/5, and IL-7delta3/4/5. Furthermore, three out-of-frame splice variants were identified, IL-7( 56bpExon2), IL-7delta4(-56bpExon2), and IL-7delta3/4/5(-56bpExon2), in which, in addition to the aforementioned exon skipping, 56 bp of the 3' end of exon 2 are omitted. Our results led us to assume that splicing-derived IL-7 isoforms play a potential role in modulating IL-7-mediated biologic effects. Further studies are required to clarify the significance of the diverse IL-7 protein isoforms for the regulation of IL-7 function and the pathogenesis of leukemia. PMID- 10386863 TI - Structure and expression of the human small cytokine B subfamily member 11 (SCYB11/formerly SCYB9B, alias I-TAC) gene cloned from IFN-gamma-treated human monocytes (THP-1). AB - Among CXC chemokines, monokine induced by interferon-gamma (IFN-gamma) (MIG) and IGN-gamma-inducible protein, 10 kDa (INP10), constitute a distinct group because of their sequence and function. We studied genomic structure and expression of a third, recently identified member of this group named small inducible cytokine B subfamily member 11 (SCYB11, formerly SCYB9B) or IFN-inducible T cell alpha chemoattractant (I-TAC). The cDNA (1445 bp) for this 94 amino acid protein (Mr 10,364) was cloned from IFN-gamma-treated human myelomonocytic cells (THP-1). The reading frame of SCYB11 is distributed to 4 exons spanning 1197 bp of the genomic sequence. In vitro transcription/translation yielded a single protein of about 10 kDa, indicating that the deduced reading frame is translated by eukaryotic ribosomes. The recombinant 73 amino acid mature protein overexpressed in Escherichia coli was chemotactic for interleukin-2 (IL-2)-selected T memory cells. Studying various cytokines and lipopolysaccharide in THP-1 cells identified IFN-gamma as the major stimulus for SCYB11 mRNA expression, followed by IFN-alpha and IFN-beta, which were about 25 times less effective. Of a panel of different human cells tested, SCYB11 mRNA was also induced in umbilical vein endothelial cells, dermal fibroblasts, and tumor cell lines from various organs, whereas it was not found in T lymphocytes activated via anti-CD3 antibodies or via IL-2. PMID- 10386864 TI - Production and in vitro characterization of recombinant chicken interleukin-2. AB - Mammalian interleukin-2 (IL-2) is a well-characterized cytokine that plays key roles in T cell differentiation and activation, B cell development, and natural killer (NK) cell stimulation. Chicken IL-2, which is the first nonmammalian IL-2 cloned, differs substantially from mammalian IL-2 molecules. We undertook to study the functions of chicken IL-2 by producing recombinant molecules in prokaryotic and eukaryotic expression systems, determining the in vitro properties of these molecules, and examining the kinetics of endogenous IL-2 production in vitro. Recombinant chicken IL-2 (rChIL-2) produced in prokaryotic and eukaryotic expression systems induced proliferation of chicken splenocytes in vitro, demonstrating that glycosylation is not required for this activity. Polyclonal antibodies generated against prokaryotically produced rChIL-2 inhibited proliferation of splenocytes induced by eukaryotically and prokaryotically produced rChIL-2, as well as endogenously produced cIL-2 obtained from ConA-stimulated splenocytes. Human IL-2 or IL-15-induced CTLL proliferation was not blocked by rChIL-2 or polyclonal anti-rChIL-2 antibodies, indicating that chicken IL-2 cannot act as an antagonist of the mammalian IL-2 response. Endogenous chicken IL-2 appears to occur in vitro as a monomer of about 14.2 kDa and is secreted within 4 h after ConA stimulation. The production of rChIL-2 provides us with a useful tool for studying avian immunology as well as a potential vaccine-enhancing agent. PMID- 10386865 TI - The IL-6/sIL-6R fusion protein hyper-IL-6 promotes neurite outgrowth and neuron survival in cultured enteric neurons. AB - The undisturbed development of the enteric nervous system depends on the supply of various neurotrophic factors during ontogenesis. Besides glial cell line derived neurotrophic factor (GDNF), leukemia inhibitory factor (LIF) and ciliary neurotrophic factor (CNTF) take part in its development. CNTF and LIF belong to the interleukin-6 (IL-6) family of cytokines. The combination of IL-6 and the soluble IL-6 receptor accelerates peripheral nerve regeneration. In this study, we examined the effect of the fusion protein Hyper-IL-6, which consists of IL-6 and the soluble receptor sIL-6R, on neurite outgrowth and neuronal survival in vitro. Myenteric plexus of newborn rats was dissected and dissociated. Cells were grown in either serum-free chemically defined medium alone or medium supplemented with sIL-6R, IL-6, sIL-6+IL-6, Hyper-IL-6, CNTF, LIF, or GDNF. Average neurite outgrowth per neuron was highest in GDNF-treated and Hyper-IL-6-treated cultures. The number of neurite-bearing neurons was reduced in GDNF cultures compared with Hyper-IL-6-treated cells, so that the total neurite outgrowth was maximal after Hyper-IL-6 stimulation. Hyper-IL-6 furthermore stimulated neuronal survival and morphologic differentiation of the enteric glia. PMID- 10386866 TI - A novel immunodeficient mouse model--RAG2 x common cytokine receptor gamma chain double mutants--requiring exogenous cytokine administration for human hematopoietic stem cell engraftment. AB - Gene transduction into immature human hematopoietic cells collected from umbilical cord blood, bone marrow, or mobilized peripheral blood cells could be useful for the treatment of genetic and acquired disorders of the hematopoietic system. Immunodeficient mouse models have been used frequently as recipients to assay the growth and differentiation of human hematopoietic stem/progenitor cells. Indeed, high levels of human cell engraftment were first reported in human/murine chimeras using NOD/SCID mice, which now are considered as the standard for these types of experiments. However, NOD/SCID mice have some clear disadvantages (including spontaneous tumor formation) that limit their general use. We have developed a new immunodeficient mouse model by combining recombinase activating gene-2 (RAG2) and common cytokine receptor gamma chain (gamma c) mutations. The RAG2-/-/gamma c- double mutant mice are completely alymphoid (T-, B-, NK-), show no spontaneous tumor formation, and exhibit normal hematopoietic parameters. Interestingly, human cord blood cell engraftment in RAG2-/-/gamma c- mice was greatly enhanced by the exogenous administration of human cytokines interleukin-(IL-3) granulocyte-macrophage colony-stimulating factor, (GM-CSF), and erythropoietin in contrast to the NOD/SCID model. This unique feature of the RAG2-/-/gamma c- mouse model should be particularly well suited for assessing the role of different cytokines in human lymphopoiesis and stem/progenitor cell function in vivo. PMID- 10386867 TI - Polymer principles and protein folding. AB - This paper surveys the emerging role of statistical mechanics and polymer theory in protein folding. In the polymer perspective, the folding code is more a solvation code than a code of local phipsi propensities. The polymer perspective resolves two classic puzzles: (1) the Blind Watchmaker's Paradox that biological proteins could not have originated from random sequences, and (2) Levinthal's Paradox that the folded state of a protein cannot be found by random search. Both paradoxes are traditionally framed in terms of random unguided searches through vast spaces, and vastness is equated with impossibility. But both processes are partly guided. The searches are more akin to balls rolling down funnels than balls rolling aimlessly on flat surfaces. In both cases, the vastness of the search is largely irrelevant to the search time and success. These ideas are captured by energy and fitness landscapes. Energy landscapes give a language for bridging between microscopics and macroscopics, for relating folding kinetics to equilibrium fluctuations, and for developing new and faster computational search strategies. PMID- 10386869 TI - The 2.1 A structure of an elicitin-ergosterol complex: a recent addition to the Sterol Carrier Protein family. AB - Elicitins, produced by most of the phytopathogenic fungi of the genus Phytophthora, provoke in tobacco both remote leaf necrosis and the induction of a resistance against subsequent attack by various microorganisms. Despite the recent description of the three-dimensional crystal structure of cryptogein (CRY), the molecular basis of the interactions between Phytophthora and plants largely remains unknown. The X-ray crystal structure, refined at 2.1 A, of a ligand complexed, mutated CRY, K13H, is reported. Analysis of this structure reveals that CRY is able to encapsulate a ligand that induces only a minor conformational change in the protein structure. The ligand has been identified as an ergosterol by gas chromatographic analysis coupled with mass spectrometry analysis. This result is consistent with biochemical data that have shown that elicitins are a distinct class of Sterol Carrier Proteins (SCP). Data presented here provide the first structural description of the pertinent features of the elicitin sterol interaction and permit a reassessment of the importance of both the key residue 13 and the mobility of the omega loop for the accessibility of the sterol to the cavity. The biological implications thereof are discussed. This paper reports the first structure of a SCP/sterol complex. PMID- 10386868 TI - Folding funnels, binding funnels, and protein function. AB - Folding funnels have been the focus of considerable attention during the last few years. These have mostly been discussed in the general context of the theory of protein folding. Here we extend the utility of the concept of folding funnels, relating them to biological mechanisms and function. In particular, here we describe the shape of the funnels in light of protein synthesis and folding; flexibility, conformational diversity, and binding mechanisms; and the associated binding funnels, illustrating the multiple routes and the range of complexed conformers. Specifically, the walls of the folding funnels, their crevices, and bumps are related to the complexity of protein folding, and hence to sequential vs. nonsequential folding. Whereas the former is more frequently observed in eukaryotic proteins, where the rate of protein synthesis is slower, the latter is more frequent in prokaryotes, with faster translation rates. The bottoms of the funnels reflect the extent of the flexibility of the proteins. Rugged floors imply a range of conformational isomers, which may be close on the energy landscape. Rather than undergoing an induced fit binding mechanism, the conformational ensembles around the rugged bottoms argue that the conformers, which are most complementary to the ligand, will bind to it with the equilibrium shifting in their favor. Furthermore, depending on the extent of the ruggedness, or of the smoothness with only a few minima, we may infer nonspecific, broad range vs. specific binding. In particular, folding and binding are similar processes, with similar underlying principles. Hence, the shape of the folding funnel of the monomer enables making reasonable guesses regarding the shape of the corresponding binding funnel. Proteins having a broad range of binding, such as proteolytic enzymes or relatively nonspecific endonucleases, may be expected to have not only rugged floors in their folding funnels, but their binding funnels will also behave similarly, with a range of complexed conformations. Hence, knowledge of the shape of the folding funnels is biologically very useful. The converse also holds: If kinetic and thermodynamic data are available, hints regarding the role of the protein and its binding selectivity may be obtained. Thus, the utility of the concept of the funnel carries over to the origin of the protein and to its function. PMID- 10386871 TI - Diversity of functions of proteins with internal symmetry in spatial arrangement of secondary structural elements. AB - We carry out a systematic analysis of the correlation between similarity of protein three-dimensional structures and their evolutionary relationships. The structural similarity is quantitatively identified by an all-against-all comparison of the spatial arrangement of secondary structural elements in nonredundant 967 representative proteins, and the evolutionary relationship is judged according to the definition of superfamily in the SCOP database. We find the following symmetry rule: a protein pair that has similar folds but belong to different superfamilies has (with a very rare exception) certain internal symmetry in its common similar folds. Possible reasons behind the symmetry rule are discussed. PMID- 10386870 TI - Identifying the structural boundaries of independent folding domains in the alpha subunit of tryptophan synthase, a beta/alpha barrel protein. AB - Two equilibrium intermediates have previously been observed in the urea denaturation of the alpha subunit of tryptophan synthase (alphaTS) from Escherichia coli, an eight-stranded beta/alpha barrel protein. In the current study, a series of amino-terminal fragments were characterized to probe the elementary folding units that may be in part responsible for this complex behavior. Stop-codon mutagenesis was used to produce eight fragments ranging in size from 105-214 residues and containing incremental elements of secondary structure. Equilibrium studies by circular dichroism indicate that all of these fragments are capable of adopting secondary structure. All except for the shortest fragment fold cooperatively. The addition of the fourth, sixth, and eighth beta-strands leads to distinct increases in structure, cooperativity, and/or stability, suggesting that folding involves the modular assembly of betaalphabeta supersecondary structural elements. One-dimensional NMR titrations at high concentrations of urea, probing the environment around His92, were also performed to test for the presence of residual structure in the fragments. All fragments that contained the first four betaalpha units of structure exhibited a cooperative unfolding transition at high concentrations of urea with significant but reduced stability relative to the full-length protein. These results suggest that the residual structure in alphaTS requires the participation of hydrophobic residues in multiple beta-strands that span the entire sequence. PMID- 10386872 TI - Sulfolobus acidocaldarius inorganic pyrophosphatase: structure, thermostability, and effect of metal ion in an archael pyrophosphatase. AB - The first crystal structure of an inorganic pyrophosphatase (S-PPase) from an archaebacterium, the thermophile Sulfolobus acidocaldarius, has been solved by molecular replacement and refined to an R-factor of 19.7% at 2.7 A. S-PPase is a D3 homohexameric protein with one Mg2+ per active site in a position similar to, but not identical with, the first activating metal in mesophilic pyrophosphatases (PPase). In mesophilic PPases, Asp65, Asp70, and Asp102 coordinate the Mg2+, while only Asp65 and Asp102 do in S-PPase, and the Mg2+ moves by 0.7 A. S-PPase may therefore be deactivated at low temperature by mispositioning a key metal ion. The monomer S-PPase structure is very similar to that of Thermus thermophilus (T-PPase) and Escherichia coli (E-PPase), root-mean-square deviations around 1 A/Calpha. But the hexamer structures of S- and T-PPase are more tightly packed and more similar to each other than they are to that of E PPase, as shown by the increase in surface area buried upon oligomerization. In T PPase, Arg116 creates an interlocking ionic network to both twofold and threefold related monomers; S-PPase has hydrophilic interactions to threefold related monomers absent in both E- and T-PPase. In addition, the thermostable PPases have about 7% more hydrogen bonds per monomer than E-PPase, and, especially in S PPase, additional ionic interactions anchor the C-terminus to the rest of the protein. Thermostability in PPases is thus due to subtle improvements in both monomer and oligomer interactions. PMID- 10386873 TI - The molecular structure of an unusual cytochrome c2 determined at 2.0 A; the cytochrome cH from Methylobacterium extorquens. AB - Cytochrome cH is the electron donor to the oxidase in methylotrophic bacteria. Its amino acid sequence suggests that it is a typical Class 1 cytochrome c, but some features of the sequence indicated that its structure might be of special interest. The structure of oxidized cytochrome cH has been solved to 2.0 A resolution by X-ray diffraction. It has the classical tertiary structure of the Class 1 cytochromes c but bears a closer gross resemblance to mitochondrial cytochrome c than to the bacterial cytochrome c2. The left-hand side of the haem cleft is unique; in particular, it is highly hydrophobic, the usual water is absent, and the "conserved" Tyr67 is replaced by tryptophan. A number of features of the structure demonstrate that the usual hydrogen bonding network involving water in the haem channel is not essential and that other mechanisms may exist for modulation of redox potentials in this cytochrome. PMID- 10386874 TI - The Schiff base complex of yeast 5-aminolaevulinic acid dehydratase with laevulinic acid. AB - The X-ray structure of the complex formed between yeast 5-aminolaevulinic acid dehydratase (ALAD) and the inhibitor laevulinic acid has been determined at 2.15 A resolution. The inhibitor binds by forming a Schiff base link with one of the two invariant lysines at the catalytic center: Lys263. It is known that this lysine forms a Schiff base link with substrate bound at the enzyme's so-called P site. The carboxyl group of laevulinic acid makes hydrogen bonds with the side chain-OH groups of Tyr329 and Ser290, as well as with the main-chain >NH group of Ser290. The aliphatic moiety of the inhibitor makes hydrophobic interactions with surrounding aromatic residues in the protein including Phe219, which resides in the flap covering the active site. Our analysis strongly suggests that the same interactions will be made by P-side substrate and also indicates that the substrate that binds at the enzyme's A-site will interact with the enzyme's zinc ion bound by three cysteines (133, 135, and 143). Inhibitor binding caused a substantial ordering of the active site flap (residues 217-235), which was largely invisible in the native electron density map and indicates that this highly conserved yet flexible region has a specific role in substrate binding during catalysis. PMID- 10386875 TI - A functional protein pore with a "retro" transmembrane domain. AB - Extended retro (reversed) peptide sequences have not previously been accommodated within functional proteins. Here, we show that the entire transmembrane portion of the beta-barrel of the pore-forming protein alpha-hemolysin can be formed by retrosequences comprising a total of 175 amino acid residues, 25 contributed by the central sequence of each subunit of the heptameric pore. The properties of wild-type and retro heptamers in planar bilayers are similar. The single-channel conductance of the retro pore is 15% less than that of the wild-type heptamer and its current-voltage relationship denotes close to ohmic behavior, while the wild type pore is weakly rectifying. Both wild-type and retro pores are very weakly anion selective. These results and the examination of molecular models suggest that beta-barrels may be especially accepting of retro sequences compared to other protein folds. Indeed, the ability to form a retro domain could be diagnostic of a beta-barrel, explaining, for example, the activity of the retro forms of many membrane-permeabilizing peptides. By contrast with the wild-type subunits, monomeric retro subunits undergo premature assembly in the absence of membranes, most likely because the altered central sequence fails to interact with the remainder of the subunit, thereby initiating assembly. Despite this difficulty, a technique was devised for obtaining heteromeric pores containing both wild-type and retro subunits. Most probably as a consequence of unfavorable interstrand side-chain interactions, the heteromeric pores are less stable than either the wild-type or retro homoheptamers, as judged by the presence of subconductance states in single-channel recordings. Knowledge about the extraordinary plasticity of the transmembrane beta-barrel of alpha-hemolysin will be very useful in the de novo design of functional membrane proteins based on the beta-barrel motif. PMID- 10386876 TI - The Ubp6 family of deubiquitinating enzymes contains a ubiquitin-like domain: SUb. AB - A sequence motif that is Similar to Ubiquitin (SUb) has been identified in the Saccharomyces cerevisiae ubiquitin-specific protease Ubp6. SUb is conserved in all known Ubp6 homologues from a spectrum of eukaryotic species and is also present in a group of hypothetical proteins of unknown function (Unk1-3) present in sequence databases. An N-terminal deletion mutant of Ubp6 that lacks SUb is still capable of cleaving alpha-linked ubiquitin fusions, suggesting that SUb forms a separate domain to the catalytic core of Ubp6 and demonstrating that it is not required for in vitro cleavage activity. A homology model of the 78 N terminal amino acids of human Ubp6, based on the known fold of ubiquitin, is presented. In human Ubp6, SUb shares only 20% sequence identity with ubiquitin. Even weaker similarity occurs between S. cerevisiae SUb and ubiquitin. The homology model supports a ubiquitin-like fold for SUb and suggests that two conserved Lys residues, corresponding to Lys48 and Lys63 of ubiquitin, are functionally important. PMID- 10386878 TI - Real-time NMR studies on a transient folding intermediate of barstar. AB - The refolding of barstar, the intracellular inhibitor of barnase, is dominated by the slow formation of a cis peptidyl prolyl bond in the native protein. The triple mutant C40/82A P27A in which two cysteine residues and one trans proline were replaced by alanine was used as model system to investigate the kinetics and structural consequences of the trans/cis interconversion of Pro48. One- and two dimensional real-time NMR spectroscopy was used to follow the trans/cis interconversion after folding was initiated by rapid dilution of the urea denatured protein. Series of 1H, 15N HSQC spectra acquired with and without the addition of peptidyl prolyl isomerase unambiguously revealed the accumulation of a transient trans-Pro48 intermediate within the dead time of the experiment. Subtle chemical shift differences between the native state and the intermediate spectra indicate that the intermediate is predominantly native-like with a local rearrangement in the Pro48 loop and in the beta-sheet region including residues Tyr47, Ala82, Thr85, and Val50. PMID- 10386877 TI - Probing the role of water in the tryptophan repressor-operator complex. AB - The Escherichia coli tryptophan repressor protein (TR) represses the transcription of several genes in response to the concentration of tryptophan in the environment. In the co-crystal structure of TR bound to a DNA fragment containing its target very few direct contacts between TR and the DNA were observed. In contrast, a number of solvent mediated contacts were apparent. NMR solution structures, however, did not resolve any solvent mediated bonds at the complex interface. To probe for the role of water in TR operator recognition, the effect of osmolytes on the interactions between TR and a target oligonucleotide bearing the operator site was examined. In the absence of specific solvent mediated hydrogen bonding interactions between the protein and the DNA, increasing osmolyte concentration is expected to strongly stabilize the TR operator interaction due to the large amount of macromolecular surface area buried upon complexation. The results of our studies indicate that xylose did not alter the binding affinity significantly, while glycerol and PEG had a small stabilizing effect. A study of binding as a function of betaine concentration revealed that this osmolyte at low concentration results in a stabilization of the 1:1 TR/operator complex, but at higher concentrations leads to a switching between binding modes to favor tandem binding. Analysis of the effects of betaine on the 1:1 complex suggest that this osmolyte has about 78% of the expected effect. If one accepts the analysis in terms of the number of water molecules excluded upon complexation, these results suggest that about 75 water molecules remain at the interface of the 1:1 dimer/DNA complex. This value is consistent with the number of water molecules found at the interface in the crystallographically determined structure and supports the notion that interfacial waters play an important thermodynamic role in the specific complexation of one TR dimer with its target DNA. However, the complexity of the effects of betaine and the small or negligible effects of the other osmolytes could also arise from osmolyte induced competition between antagonistic coupled reactions. PMID- 10386879 TI - Study of the stability and unfolding mechanism of BBA1 by molecular dynamics simulations at different temperatures. AB - BBA1 is a designed protein that has only 23 residues. It is the smallest protein without disulfide bridges that has a well-defined tertiary structure in solution. We have performed unfolding molecular dynamics simulations on BBA1 and some of its mutants at 300, 330, 360, and 400 K to study their kinetic stability as well as the unfolding mechanism of BBA1. It was shown that the unfolding simulations can provide insights into the forces that stabilize the protein. Packing, hydrophobic interactions, and a salt bridge between Asp12 and Lys16 were found to be important to the protein's stability. The unfolding of BBA1 goes through two major steps: (1) disruption of the hydrophobic core and (2) unfolding of the helix. The beta-hairpin remains stable in the unfolding because of the high stability of the type II' turn connecting the two beta-strands. PMID- 10386880 TI - The 80s loop of the catalytic chain of Escherichia coli aspartate transcarbamoylase is critical for catalysis and homotropic cooperativity. AB - The X-ray structure of the Escherichia coli aspartate transcarbamoylase with the bisubstrate analog phosphonacetyl-L-aspartate (PALA) bound shows that PALA interacts with Lys84 from an adjacent catalytic chain. To probe the function of Lys84, site-specific mutagenesis was used to convert Lys84 to alanine, threonine, and asparagine. The K84N and K84T enzymes exhibited 0.08 and 0.29% of the activity of the wild-type enzyme, respectively. However, the K84A enzyme retained 12% of the activity of the wild-type enzyme. For each of these enzymes, the affinity for aspartate was reduced 5- to 10-fold, and the affinity for carbamoyl phosphate was reduced 10- to 30-fold. The enzymes K84N and K84T exhibited no appreciable cooperativity, whereas the K84A enzyme exhibited a Hill coefficient of 1.8. The residual cooperativity and enhanced activity of the K84A enzyme suggest that in this enzyme another mechanism functions to restore catalytic activity. Modeling studies as well as molecular dynamics simulations suggest that in the case of only the K84A enzyme, the lysine residue at position 83 can reorient into the active site and complement for the loss of Lys84. This hypothesis was tested by the creation and analysis of the K83A enzyme and a double mutant enzyme (DM) that has both Lys83 and Lys84 replaced by alanine. The DM enzyme has no cooperativity and exhibited 0.18% of wild-type activity, while the K83A enzyme exhibited 61% of wild-type activity. These data suggest that Lys84 is not only catalytically important, but is also essential for binding both substrates and creation of the high-activity, high-affinity active site. Since low-angle X-ray scattering demonstrated that the mutant enzymes can be converted to the R-structural state, the loss of cooperativity must be related to the inability of these mutant enzymes to form the high-activity, high-affinity active site characteristic of the R-functional state of the enzyme. PMID- 10386881 TI - The paradox between m values and deltaCp's for denaturation of ribonuclease T1 with disulfide bonds intact and broken. AB - Urea-induced denaturations of RNase T1 and reduced and carboxyamidated RNase T1 (RTCAM) as a function of temperature were analyzed using the linear extrapolation method, and denaturation m values, deltaCp, deltaH, deltaS, and deltaG quantities were determined. Because both deltaCp and m values are believed to reflect the protein surface area newly exposed on denaturation, the prediction is that the ratio of m values for RNase T1 and RTCAM should equal the deltaCp ratio for the two proteins. This is not the case, for it is found that the m value of RTCAM is 1.5 times that of RNase T1, while the denaturation deltaCp's for the two proteins are identical. The paradox of why the two parameters, m and deltaCp, are not equivalent in their behavior is of importance in the interpretations of their respective molecular-level meanings. It is found that the measured denaturation deltaCp's are consistent with deltaCp's calculated on the basis of empirical relationships between the change in surface area on denaturation (deltaASA), and that the measured m value of RNase T1 agrees with m calculated from empirical data relating m to deltaASA. However, the measured m of RTCAM is so much out of line with its calculated m as to call into question the validity of always equating m with surface area newly exposed on denaturation. PMID- 10386882 TI - Autonomous folding of a peptide corresponding to the N-terminal beta-hairpin from ubiquitin. AB - The N-terminal 17 residues of ubiquitin have been shown by 1H NMR to fold autonomously into a beta-hairpin structure in aqueous solution. This structure has a specific, native-like register, though side-chain contacts differ in detail from those observed in the intact protein. An autonomously folding hairpin has previously been identified in the case of streptococcal protein G, which is structurally homologous with ubiquitin, but remarkably, the two are not in topologically equivalent positions in the fold. This suggests that the organization of folding may be quite different for proteins sharing similar tertiary structures. Two smaller peptides have also been studied, corresponding to the isolated arms of the N-terminal hairpin of ubiquitin, and significant differences from simple random coil predictions observed in the spectra of these subfragments, suggestive of significant limitation of the backbone conformational space sampled, presumably as a consequence of the strongly beta-structure favoring composition of the sequences. This illustrates the ability of local sequence elements to express a propensity for beta-structure even in the absence of actual sheet formation. Attempts were made to estimate the population of the folded state of the hairpin, in terms of a simple two-state folding model. Using published "random coil" values to model the unfolded state, and values derived from native ubiquitin for the putative unique, folded state, it was found that the apparent population varied widely for different residues and with different NMR parameters. Use of the spectra of the subfragment peptides to provide a more realistic model of the unfolded state led to better agreement in the estimates that could be obtained from chemical shift and coupling constant measurements, while making it clear that some other approaches to population estimation could not give meaningful results, because of the tendency to populate the beta-region of conformational space even in the absence of the hairpin structure. PMID- 10386883 TI - A novel recombinant single-chain hepatitis C virus NS3-NS4A protein with improved helicase activity. AB - Hepatitis C virus (HCV) nonstructural protein 3 (NS3) has been shown to possess protease and helicase activities and has also been demonstrated to spontaneously associate with nonstructural protein NS4A (NS4A) to form a stable complex. Previous attempts to produce the NS3/NS4A complex in recombinant baculovirus resulted in a protein complex that aggregated and precipitated in the absence of nonionic detergent and high salt. A single-chain form of the NS3/NS4A complex (His-NS4A21-32-GSGS-NS3-631) was constructed in which the NS4A core peptide is fused to the N-terminus of the NS3 protease domain as previously described (Taremi et al., 1998). This protein contains a histidine tagged NS4A peptide (a.a. 21-32) fused to the full-length NS3 (a.a. 3-631) through a flexible tetra amino acid linker. The recombinant protein was expressed to high levels in Escherichia coli, purified to homogeneity, and examined for NTPase, nucleic acid unwinding, and proteolytic activities. The single-chain recombinant NS3-NS4A protein possesses physiological properties equivalent to those of the NS3/NS4A complex except that this novel construct is stable, soluble and sixfold to sevenfold more active in unwinding duplex RNA. Comparison of the helicase activity of the single-chain recombinant NS3-NS4A with that of the full-length NS3 (without NS4A) and that of the helicase domain alone suggested that the presence of the protease domain and at least the NS4A core peptide are required for optimal unwinding activity. PMID- 10386884 TI - The crystal structure of a bacterial, bifunctional 5,10 methylene tetrahydrofolate dehydrogenase/cyclohydrolase. AB - The structure of a bifunctional 5,10-methylene-tetrahydrofolate dehydrogenase/cyclohydrolase from Escherichia coli has been determined at 2.5 A resolution in the absence of bound substrates and compared to the NADP-bound structure of the homologous enzyme domains from a trifunctional human synthetase enzyme. Superposition of these structures allows the identification of a highly conserved cluster of basic residues that are appropriately positioned to serve as a binding site for the poly-gamma-glutamyl tail of the tetrahydrofolate substrate. Modeling studies and molecular dynamic simulations of bound methylene tetrahydrofolate and NADP shows that this binding site would allow interaction of the nicotinamide and pterin rings in the dehydrogenase active site. Comparison of these enzymes also indicates differences between their active sites that might allow the development of inhibitors specific to the bacterial target. PMID- 10386886 TI - Simple sequence is abundant in eukaryotic proteins. AB - All proteins of Saccharomyces cerevisiae have been compared to determine how frequently segments from one protein are present in other proteins. Proteins that are recently evolutionarily related were excluded. The most frequently present protein segments are long, tandem repetitions of a single amino acid. For some of these segments, up to 14% of all proteins in the genome were found to have similar peptides within them. These peptide segments may not be functional protein domains. Although they are the most common shared feature of yeast proteins, their ubiquity and simplicity argue that their probable function may be to simply serve as spacers between other protein motifs. PMID- 10386885 TI - Formation of amyloid fibrils by peptides derived from the bacterial cold shock protein CspB. AB - Three peptides covering the sequence regions corresponding to the first two (CspB 1), the first three (CspB-2), and the last two (CspB-3) beta-strands of CspB, the major cold shock protein of Bacillus subtilis, have been synthesized and analyzed for their conformations in solution and for their precipitation behavior. The peptides are nearly insoluble in water, but highly soluble in aqueous solutions containing 50% acetonitrile (pH 4.0). Upon shifts of the solvent condition toward lower or higher acetonitrile concentrations, the peptides all form fibrils resembling those observed in amyloid associated diseases. These fibrils have been identified and characterized by electron microscopy, binding of the dye congo red, and X-ray fiber diffraction. Characterization of the peptides in solution by circular dichroism and NMR spectroscopy shows that the formation of these fibrils does not require specific preformed secondary structure in the solution state species. While the majority of the soluble fraction of each peptide is monomeric and unstructured, different types of structures including alpha-helical, beta sheet, and random coil conformations are observed under conditions that eventually lead to fibril formation. We conclude that the absence of tertiary contacts under solution conditions where binding interactions between peptide units are still favorable is a crucial requirement for amyloid formation. Thus, fragmentation of a sequence, like partial chemical denaturation or mutation, can enhance the capacity of specific protein sequences to form such fibrils. PMID- 10386887 TI - New insight into the pH-dependent conformational changes in bovine beta lactoglobulin from Raman optical activity. AB - We have studied the conformation of beta-lactoglobulin in aqueous solution at room temperature over the pH range approximately 2.0-9.0 using vibrational Raman optical activity (ROA). The ROA spectra clearly show that the basic up and down beta-barrel core is preserved over the entire pH range, in agreement with other studies. However, from the shift of a sharp positive ROA band at approximately 1268 to approximately 1294 cm(-1) on going from pH values below that of the Tanford transition, which is centered at pH approximately 7.5, to values above, the Tanford transition appears to be associated with changes in the local conformations of residues in loop sequences possibly corresponding to a migration into the alpha-helical region of the Ramachandran surface from a nearby region. These changes may be related to those detected in X-ray crystal structures which revealed that the Tanford transition is associated with conformational changes in loops which form a doorway to the interior of the protein. The results illustrate how the ability of ROA to detect loop and turn structure separately from secondary structure is useful for studying conformational plasticity in proteins. PMID- 10386888 TI - Selective association of protein molecules followed by mass spectrometry. AB - Nanoflow electrospray mass spectrometry was used to monitor the formation of protein heterodimers of HU proteins from Bacillus stearothermophilus and Bacillus subtilis. This has enabled us to analyze both thermodynamic and kinetic features associated with the dissociation of homodimeric HU proteins. The results obtained correlate well with the kinetics of the protein dissociation process and the free energy difference between homo- and heterodimeric species anticipated from other studies. We suggest that this approach will have general applicability in studying protein association and dissociation under near-equilibrium conditions and will be relevant to a wide range of biological systems. PMID- 10386889 TI - Simple synthesis of sialyllactose-carrying polystyrene and its binding with influenza virus. AB - Glycoconjugate polystyrenes bearing sialyllactose moieties were prepared via a simple method from a mixture of alpha2-6 and a2-3 linked sialyllactose isomers of bovine milk origin. The reducing end of sialyllactose was converted to an amino function with ammonium hydrogen carbonate and then coupled with p-vinylbenzoyl chloride. The resulting styrene derivative substituted with sialyllactose via an amide linkage was polymerized with ammonium peroxodisulfate and N,N,N',N tetramethylethylenediamine in water at 30 degrees C. The interaction of the glycopolymer with influenza A and B viruses was investigated by three different methods. The glycopolymer inhibited the hemagglutination of influenza A virus (PR/8/34) and its activity was 10(3) times higher than that of the oligosaccharide itself. The cytopathic effect of virus-infected MDCK (Madine Darby canine kidney) cells was inhibited by the glycopolymer. The homopolymer showed 10(2) times higher inhibitory activity than naturally-occurring fetuin. It was also found that various viruses could be trapped by the glycopolymer adsorbed on a polystyrene surface. The inhibitory and trapping activities of the glycopolymers were correlated with the sialyl linkage specificities of the virus strains. PMID- 10386891 TI - Characterization of a low-sulfated chondroitin sulfate from the body of Viviparus ater (mollusca gastropoda). Modification of its structure by lead pollution. AB - A chondroitin sulfate was purified from the body of Viviparus ater(Mollusca gastropoda) and analyzed for molecular mass, constituent disaccharides, and structure by 1H NMR and 1H 2D NMR. A quite unique glycosaminoglycan species was isolated having a high molecular mass (greater than 45,000) and low charge density, about 0.60, due to the presence of 42% non-sulfated disaccharide, 5% 6 sulfated disaccharide, 48% 4-sulfated disaccharide, and 5% 4,6-disulfated disaccharide. Specimens of Mollusca were also submitted to lead exposure for different times, and the effect on chondroitin sulfate structure was studied. After 96 h treatment a strong decrease in chondroitin sulfate content was observed with a significant modification of its structure producing a more desulfated polymer, in particular in position 4 of the galactosamine unit. Simultaneously, the amount of unsaturated non-sulfated disaccharide increased with an overall decrease of the charge density. PMID- 10386890 TI - Structural analysis of N-glycans from allergenic grass, ragweed and tree pollens: core alpha1,3-linked fucose and xylose present in all pollens examined. AB - The N-glycans from soluble extracts of ten pollens were examined. The pyridylaminated oligosaccharides derived from these sources were subject to gel filtration and reverse-phase HPLC, in conjunction with exoglycosidase digests, and in some cases matrix-assisted laser desorption-ionisation mass spectrometry. In comparison to known structures, it was possible to determine the major structures of the N-glycans derived from Kentucky blue grass (Poa pratensis), rye (Secale cerale), ryegrass (Lolium perenne), short ragweed (Ambrosia elatior), giant ragweed (Ambrosia trifida), birch (Betula alba), hornbeam (Carpinus betulus), horse chestnut (Aesculus hippocastanum), olive (Olea europaea) and snake-skin pine (Pinus leucodermis) pollen extracts. For grass pollens the major glycans detected were identical in properties to: [structure in text] Grass pollens also contained some minor structures with one or two non-reducing terminal N-acetylglucosamine residues. In the ragweed pollens, the major structures carried core alpha1,3-linked fucose with or without the presence of xylose. In tree pollen extracts, the major structures were either xylosylated, with or without fucose and terminal N-acetylglucosamine residues, with also significant amounts of oligomannose structures. These results are compatible with the hypothesis that the carbohydrate structures are another potential source of immunological cross-reaction between different plant allergens. PMID- 10386892 TI - Panosialins, inhibitors of an alpha1,3-fucosyltransferase Fuc-TVII, suppress the expression of selectin ligands on U937 cells. AB - Panosialins A and B were isolated as inhibitors of an alpha1,3 fucosyltransferase, Fuc-TVII, which is a key enzyme in the biosynthesis of selectin ligands, from culture broth of Streptomyces sp. Panosialins A and B inhibited the Fuc-TVII activity with IC50 values of 4.8 and 5.3 microg/ml, respectively. Panosialin A suppressed expression of selectin ligands on U937 cells, and inhibited the cell adhesion to immobilized E-selectin-immunoglobulin. Panosialins are the first reported Fuc-TVII inhibitors which can suppress the biosynthesis of selectin ligands and then inhibit selectin-mediated cell adhesion. PMID- 10386893 TI - Lectin histochemistry study in the human vas deferens. AB - The oligosaccharide sequences of glycoconjugates in the normal human vas deferens and the nature of the saccharide linkage were studied by lectin histochemistry. The cytoplasm of all epithelial cell types (principal cells, basal cells, and mitochondria-rich cells) and luminal contents reacted positively with WGA, MAA, PNA, DSA, LTA, UEA-I, AAA, and ConA. The reaction was more intense in the stereocilia of principal cells. Cytoplasmic staining was diffuse except for PNA and DSA labeling which was limited to the apical cytoplasm and stereocilia of columnar cells. The cytoplasm of all cell types also reacted diffusely with HPA, although staining was weak and was not observed in the stereocilia. Positive reaction with SBA only was encountered in the stereocilia of principal cells. SNA, LTA, and DBA were unreactive. GNA-labeling showed a granular distribution in the supranuclear cytoplasm of columnar epithelial cells. Reactions with MAA, PNA, DSA, AAA, HPA and SBA disappeared after the beta-elimination reaction. Reactions with WGA and UEA-I decreased after beta-elimination or Endo-F digestion. Reactions with ConA and GNA were suppressed by Endo-F digestion. Reactions with PNA, HPA, and SBA increased after desialylation. Of all the lectins that label the luminal contents of the vas deferens, only UEA-I was not found in the luminal contents of seminiferous tubules and epididymis and, thus, this lectin would probably bind to glycoproteins secreted by the vas deferens. The chemical treatments used suggest that this secretion contains fucose residues located in both N- and O-linked oligosaccharides. The other lectins may label secreted proteins, but also structural proteins or proteins reabsorbed from the luminal fluid. The lectin- binding pattern of mitochondria-rich cells in the vas deferens differed from that found in the epididymis. PMID- 10386894 TI - B lymphocyte galactosyltransferase protein levels in normal individuals and in patients with rheumatoid arthritis. AB - We have quantified the level of beta4-galactosyltransferase protein in human B lymphocytes using an ELISA-based assay. Between 1-10ng of beta4 galactosyltransferase was detected per mg total cellular protein, indicating that this enzyme constitutes <0.001% of B lymphocyte cellular protein. Akin to previous studies, individuals with rheumatoid arthritis exhibited reduced lymphocytic galactosyltransferase enzyme activity compared with normal controls when using ovalbumin as the acceptor substrate. The levels of enzyme protein present in B lymphocytes from patients with rheumatoid arthritis was, however, not reduced suggesting that the B lymphocyte galactosyltransferase catalytic activity may be regulated post-translationally. PMID- 10386895 TI - Synthetic sialylphosphatidylethanolamine derivatives bind to human influenza A viruses and inhibit viral infection. AB - We synthesized the sialylphosphatidylethanolamine (sialyl PE) derivatives Neu5Ac PE, (Neu5Ac)2-PE, Neu5Ac-PE (amide) and Neu5Ac-PE (methyl). We examined the anti viral effects of the derivatives on human influenza A virus infection by ELISA/virus-binding, hemagglutination inhibition, hemolysis inhibition and neutralization assays. The sialyl PE derivatives that we examined bound to A/Aichi/2/68, A/Singapore/1/57 and A/Memphis/1/71 strains of H3N2 subtype, but not to A/PR/8/34 strain of H1N1 subtype. The derivatives inhibited viral hemagglutination and hemolysis of human erythrocytes with A/Aichi/2/68 and A/Singapore/1/57 (H3N2), but not with A/PR/8/34 (H1N1). The inhibitory activity of the (Neu5Ac)2-PE derivative was the strongest of all sialyl PE derivatives (IC50, 35 microM to 40 microM). Sialyl PE derivatives also inhibited the infection of A/Aichi/2/68 in MDCK cells. Complete inhibition was observed at a concentration between 0.3 to 1.3 mM. IC50 of (Neu5Ac)2-PE was 15 microM in A/Aichi/2/68 strain. Taken together, the synthetic sialyl PE derivatives may be effective reagents against infection of some types of influenza A viruses. PMID- 10386897 TI - Lack of influence of hysterectomy on meal size and meal number in Fischer-344 rats. AB - Based on our previous observation that, when eating the same amount of food per 100 g b.wt., male rats gain five to seven times more weight than females who have an estrous cycle every 4 to 5 days, we questioned whether lower weight gain seen in female rats could be the result of increased energy cost in preparing endometrium for anticipated fertilization. Because the uterus modulates estrogenic effects on other hormone-dependent behaviors, for example, sexual receptivity and lordosis, we performed this study to determine if estrogen mediated cyclical changes in food intake and feeding pattern occur after hysterectomy. Fifteen female Fischer 344 rats were randomized during the estrous phase to either hysterectomy with ovarian preservation or sham operation. A rat eater meter was used to continuously measure food intake, meal number, and meal size for two estrous cycles before and four cycles after surgery. Both groups showed the estrous phase linked cycling in meal number, meal size, and food intake. No differences existed between the two groups in these indices either before or after surgery. No differences existed between groups in rate of body weight gain after surgery, 0.95 +/- 0.13 g/day in hysterectomized and 0.77 +/- 0.1 g/day in sham-operated rats. We conclude that hysterectomy has no effect on rate of weight gain, food intake, and estrus linked cyclical feeding pattern in Fischer 344 rats. PMID- 10386898 TI - Influence of age on behavioural response in the light/dark paradigm. AB - We have compared the performance of male Swiss mice at different ages (correlated with different body weight; 12-34 g) in the light/dark test of anxiety. Mice received saline only. The best age at which control values were optimum was that of 4 weeks old. Mice at this age spent 58% of the total test duration in the dark compartment. The oldest mice (i.e., 8 weeks old) exhibited an increase in total activity characterised by increase in movements in each compartment, together with an increase in the number of transitions. An age-related effect was found suggesting caution when interpreting the results of mice in the light/dark paradigm, the best period being that of 4 weeks. PMID- 10386899 TI - Fetal alcohol exposure blocks full masculinization of the dorsolateral nucleus in rat spinal cord. AB - Male rats prenatally exposed to a combination of stress and ethanol show severely impaired ejaculatory patterns. This study examined two sexually dimorphic nuclei in the lumbar spinal cord implicated in the control of male copulatory reflexes in rats whose mothers were exposed to alcohol, to stress, or to both treatments during pregnancy. Alcohol exposure led to a marked decrease (22%) in the number of motor neurons in the dorsolateral nucleus (DLN) of the adult male offspring, but no significant change in cell count was detectable in the sexually dimorphic nucleus of the bulbocavernosus (SNB). The combination of alcohol and stress did not enhance the effect on the DLN above that produced by alcohol alone. Somal sizes in the DLN and SNB were not altered by any of the treatment conditions. Alcohol exposure probably leads to incomplete masculinization of the DLN in male rats by decreasing testicular steroidogenesis during the fetal stage(s) when sexual differentiation is ongoing in that CNS structure. PMID- 10386896 TI - Group and individual gustatory reaction times and Pieron's law. AB - Simple reaction times (SRT) to eight substances belonging to the four classical taste families were evaluated. The same eight subjects participated in all experiments. The functional relationship between SRT and concentration for group and for individual data were examined. Equations presented by different authors to describe RT data are discussed. The Pieron function [(SRT - t0) = betaI-alpha] best fits the gustatory data collected in the present experiments. These results, together with others taken from previous studies, show that the exponent of salt and acid taste functions is lower than 1.0 with a relatively short t0. Sweet and bitter exponents were equal t0 or higher than 1, with a larger t0. Individual performances correlated with taste families for salt and acid. However, the limited samples of some solutions sets some limits to the interpretation of RT to taste substances. PMID- 10386900 TI - Protective effect of acidic fibroblast growth factor against ischemia-induced learning and memory deficits in two tasks in gerbils. AB - The influence of transient forebrain ischemia on behavioral performance, and the effect of intracerebroventricular (i.c.v.) injection of acidic fibroblast growth factor (aFGF) on such ischemia-induced deficits were examined in Mongolian gerbils by assessing learning and memory in two tasks: passive avoidance and Morris water maze. A 5-min period of forebrain ischemia led to learning and memory deficits in both tasks, and also to neuronal death in the hippocampal CA1 region. Continuous i.c.v. infusion of aFGF bilaterally into the lateral ventricules by osmotic minipumps over 2 days before, and 5 days after the ischemia (a total of 3.6 microg/gerbil) largely prevented both the ischemia induced behavioral deficits and the neuronal death in the hippocampus. These observations suggest that the hippocampus is a critical site for the performance of the two tasks, and that aFGF has a protective effect against such ischemia induced learning and memory deficits in gerbils. PMID- 10386901 TI - Thermal preference behavior following clonidine, norepinephrine, isoproterenol, and ephedrine. AB - A thermal gradient (temperature range 7-45 degrees C) was used to assess ambient temperature (Ta) preferences of rats following treatment with clonidine (25 microg/kg), norepinephrine (NE, 250 microg/kg), isoproterenol (ISO, 50 microg/kg), and ephedrine (EPH, 10 mg/kg). Clonidine produced a preference for a temperature (31.5 degrees C) slightly warmer than that preferred after saline (28.3 degrees C), but this resulted in no significant change in posttest colonic temperature (Tc). NE, ISO and EPH produced a preference for a colder region of the gradient (20-22 degrees C) compared to saline (24.5-28.9 degrees C). Posttest Tc was reduced significantly from 37.7-37.9 degrees C after saline to 37.2 degrees C (NE), 37.3 degrees C (ISO), and 36.8 degrees C (EPH). Thus, given the opportunity to select an environmental temperature, the animals selected a Ta that resulted in significantly lower body temperatures after NE, ISO, and EPH. This suggests that paradoxical thermoregulatory effects of these thermogenic adrenergic agonists are due, at least in part, to a preference for a lower body temperature. PMID- 10386902 TI - Effects of mating on c-fos expression in the brains of male macaques. AB - The c-fos polyclonal anti-c-fos antibody was used to examine the effects of mating on Fos expression in brain neurons of 11 male macaques. Behavior tests were for 30 min, five males were unmated, four were mated, and two were social controls. Mated males were killed 60 min after ejaculation. Social controls were paired with females, but mating did not occur. Fos immunoreactive (Fos-ir) neuronal nuclei were counted in nine brain regions extending from the medial preoptic to the mammillary body area of all males. In contrast to previous reports on nonprimate laboratory species, overall there was as much Fos-ir in unmated as in mated males. Moreover, there was significantly less Fos expression in four brain regions (known to contain steroid receptors), namely, ventromedial hypothalamus, arcuate nucleus, lateral mammillary area, and bed nucleus of stria terminalis, of mated than of unmated males. There were no significant differences between mated and unmated males in the 5 other brain regions studied. These findings may reflect taxonomic differences between primates and nonprimates, or result from greater neural activation in feral animals maintained in a laboratory than in domesticated, inbred laboratory species. The simplest interpretation would be that neural activity in the male primate is turned off by mating in some brain sites but not in others. PMID- 10386903 TI - Temporal and behavioral patterning of parturition in rabbits and rats. AB - Although the rabbit (Oryctolagus cuniculus) continues to play an important role in the study of parturitional processes, a detailed behavioral description of birth in this species, necessary for accurately assessing the effects of experimental manipulation, is lacking. It is the aim of this report to provide such a description and to compare it with corresponding behavior in the better studied rat. Ten pregnant chinchilla-breed rabbits and 10 pregnant Wistar rats were placed in glass-bottomed observation cages 2 days before term, and their behavior recorded on closed-circuit video, viewing the animals from below. All aspects of parturition were accomplished much faster in rabbits than rats; latency to birth of first pup, rate of delivery, duration of vaginal retention, time spent by mothers eating placentas, and in licking and nursing pups. In contrast to rat pups, rabbits were usually born separated from the placenta and already free from membranes. They were much more active, and well able to cast off any remaining membranes, suckle, and survive, whether directly attended to by the mother or not. We conclude that the tight temporal organization of events in the rabbit provides an unusually sensitive assay for investigating mechanisms underlying mammalian parturition. PMID- 10386904 TI - Daily activity and body temperature rhythms do not change simultaneously with age in laboratory mice. AB - Daily rhythms of locomotor activity (AR) and body temperature (TR) were investigated in juvenile, adult, and senile female laboratory mice (5, 16, or 65 weeks old). All daily patterns were bimodal, with a main maximum in the dark and a secondary one immediately following lights on. The juvenile mice showed the highest magnitude of oscillation of the AR but the lowest magnitude of the TR; the magnitudes of the TR of adult and senile animals were not different, whereas those of AR in senile mice approached zero. For the AR, but not the TR, a phase advance with age was observed. The effect of locomotor activity on the body temperature was higher during the light time (minimum of motor activity) than during the dark time (maximum activity), and was least in juvenile mice. The calculated daily temperatures corresponding to zero activity gave rhythms that showed no age-dependent differences in daily mean or magnitude. This implies that the age-dependent changes of the TR were due mainly to masking effects. PMID- 10386905 TI - Endocrine and psychophysiological aspects of human adaptation to the extreme. AB - Human beings need to adapt to any extreme, unknown, or isolated environment. This adaptation requires changes in the normal regulation of psychophysiological homeostasis, as described in terms of stress reaction. The aim of the present study was to monitor the processes of human adaptation to cold and isolated areas in Antarctica during the 12th expedition of the Italian National Research Program. Nine healthy subjects (experimental subjects), members of the expedition, and nine controls in Italy, were studied over a period of 2 months. Anterior pituitary hormone secretion, insulin, and melatonin, plus routine blood test, blood pressure, and ECG were performed. In addition, psychophysiological correlates were also recorded before and after the expedition period. In experimental subjects results of metabolic data suggested the presence of an increased peripheral insulin sensitivity at the end of the permanence in the station and a significant increased of total cholesterol. Hematocrit also significantly increased due to the conditions of hypobaric hypoxia. Results of endocrine data showed a significant decrease (p < 0.05) of hormone levels, which was associated with a significant decrement of the Galvanic Skin Response (GSR) activity to a standardized cognitive stress. No significant differences were reported in the controls. The data suggest that the exposure to the extreme environment develops a possible psychophysiological mechanism(s) that decreases the individual arousal. PMID- 10386906 TI - Differential satiating effects of fats in the small intestine of obesity resistant and obesity-prone rats. AB - The effects of duodenal infusions of fats on sham feeding was measured in two strains of rats that differ in their susceptibility to fat-induced obesity. Osborne-Mendel rats are prone to developing obesity on a high-fat diet and preferentially choose fats over carbohydrates in macronutrient selection paradigms. In contrast, S 5B/PL rats are resistant to developing obesity when eating a high-fat diet, and preferentially choose carbohydrates in macronutrient selection paradigms. To test the hypothesis that differences in the satiating potency of fats in the small intestine contributed to these differences between the two strains, we measured the effects of duodenal infusions of Intralipid and sodium linoleate on sham-feeding intakes. The results were consistent with the hypothesis. Duodenal infusions of either of these fats decreased intake significantly more in S5B/PL rats than in Osborne-Mendel rats. Both rat strains sham fed similar amounts when intestinally infused with 0.15 M NaCl. These results suggest that differences in responses to intestinal satiating mechanisms may contribute to the differences in susceptibility to fat-induced obesity in these rat strains. PMID- 10386908 TI - High-frequency ultrasonic vocalizations index conditioned pharmacological reward in rats. AB - We have proposed that short (<0.5 s), high-frequency (approximately 50 kHz) ultrasonic vocalizations ("50-kHz USVs") index a positive affective state in adult rats, because they occur prior to rewarding social interactions (i.e., rough-and-tumble play, sex). To evaluate this hypothesis in the case of nonsocial stimuli, we examined whether rats would make increased 50-kHz USVs in places associated with the administration of rewarding pharmacological compounds [i.e., amphetamine (AMPH) and morphine (MORPH)]. In Experiment 1, rats made a greater percentage of 50-kHz USVs on the AMPH-paired side of a two-compartment chamber than on the vehicle-paired side, even after statistical correction for place preference. In Experiment 2, rats made a higher percentage of 50-kHz USVs on the MORPH-paired side than on the vehicle-paired side, despite nonsignificant place preference. These findings support the hypothesis that 50-kHz USVs mark a positive affective state in rats and introduce a novel and rapid marker of pharmacological reward. PMID- 10386907 TI - Long-term behavioral effects of repetitive pain in neonatal rat pups. AB - Human preterm neonates are subjected to repetitive pain during neonatal intensive care. We hypothesized that exposure to repetitive neonatal pain may cause permanent or long-term changes because of the developmental plasticity of the immature brain. Neonatal rat pups were stimulated one, two, or four times each day from P0 to P7 with either needle prick (noxious groups N1, N2, N4) or cotton tip rub (tactile groups T1, T2, T4). In groups N2, N4, T2, T4 stimuli were applied to separate paws at hourly intervals;each paw was stimulated only once a day. Identical rearing occurred from P7 to P22 days. Pain thresholds were measured on P16, P22, and P65 (hot-plate test), and testing for defensive withdrawal, alcohol preference, air-puff startle, and social discrimination tests occurred during adulthood. Adult rats were exposed to a hot plate at 62 degrees C for 20 s, then sacrificed and perfused at 0 and 30 min after exposure. Fos expression in the somatosensory cortex was measured by immunocytochemistry. Weight gain in the N2 group was greater than the T2 group on P16 (p < 0.05) and P22 (p < 0.005); no differences occurred in the other groups. Decreased pain latencies were noted in the N4 group [5.0 +/- 1.0 s vs. 6.2 +/- 1.4 s on P16 (p < 0.05); 3.9 +/- 0.5 s vs. 5.5 +/- 1.6 s on P22 (p < 0.005)], indicating effects of repetitive neonatal pain on subsequent development of the pain system. As adults, N4 group rats showed an increased preference for alcohol (55 +/- 18% vs. 32 +/- 21%; p = 0.004); increased latency in exploratory and defensive withdrawal behavior (p < 0.05); and a prolonged chemosensory memory in the social discrimination test (p < 0.05). No significant differences occurred in corticosterone and ACTH levels following air-puff startle or in pain thresholds at P65 between N4 and T4 groups. Fos expression at 30 min after hot-plate exposure was significantly greater in all areas of the somatosensory cortex in the T4 group compared with the N4 group (p < 0.05), whereas no differences occurred just after exposure. These data suggest that repetitive pain in neonatal rat pups may lead to an altered development of the pain system associated with decreased pain thresholds during development. Increased plasticity of the neonatal brain may allow these and other changes in brain development to increase their vulnerability to stress disorders and anxiety-mediated adult behavior. Similar behavioral changes have been observed during the later childhood of expreterm neonates who were exposed to prolonged periods of neonatal intensive care. PMID- 10386909 TI - Inhibition of food intake by CRF in chickens. AB - The effect of intracerebroventricular (i.c.v.) injection of corticotrophin releasing factor (CRF) on food and water intake and on body temperature in chickens was determined. Both broiler and Leghorn type chickens were utilized in this experiment. A stainless steel guide cannula was surgically implanted into the right lateral ventricle of each bird. The i.c.v. injection of CRF significantly decreased food intake in both fed and overnight-fasted broilers and Leghorns. Water intake was decreased by CRF in Leghorns but not broilers. When CRF was injected into Leghorns given access to water, but not food, water intake was not affected. Body temperature was not affected by the i.c.v. injection of CRF. These results suggest that CRF acts within the central nervous system of chickens to decrease food intake while having no affect on water intake or body temperature. PMID- 10386910 TI - Age-induced cognitive alterations in OF1 mice. AB - Female OF1 mice aged 17-18 months were compared with female OF1 mice aged 7-11 weeks for locomotor activity, pain sensitivity, and cognitive performance using the Morris water maze, passive and active avoidance, and the elevated plus-maze learning protocol. Performance of old mice was impaired compared to those of young mice for both locomotor activity, pain sensitivity, and the four cognitive tests including the elevated plus-maze not previously used in studies on aging. Using complementary experiments and a detailed analysis of the results, we have shown that the reduction of learning and memory do not result from a decline of sensory and motor capacities. We conclude that female OF1 mice aged 17-18 months show true cognitive deficits. PMID- 10386911 TI - Control of reproductive and energetic status by environmental cues in a desert rodent, Shaw's jird. AB - The photoperiod is the controller of reproductive cycles in temperate climates for most mammalian species. Several nonphotoperiodic cues appear to control reproductive status at lower latitudes. We tested the roles of the photoperiod or water availability on the reproductive status of the desert-dwelling Shaw's jird (Meriones shawi) trapped from a moderately temperate climate (approximately 30 degrees N in Egypt). Males and females were transported to the laboratory and, in Experiment 1, were housed in either the longest (LDs) or shortest (SDs) photoperiod that occurs naturally at this latitude (14 h light, 10 h dark, and 10 h light, 14 h dark, respectively). In Experiment 2, LD-housed male jirds were subjected to a water availability schedule that inhibits reproductive status in a closely related species (Meriones unguiculatus). Specifically, one group had no free water, but had lettuce available once a week for 24 h (control jirds received free water for 10-60 min/day). Neither photoperiod nor free-water deprivation affected reproductive status of male or female jirds. That is, neither testes mass nor spermatogenetic activity (males), nor uterine mass nor folliculogenesis (females) were affected by either condition. In addition, photoperiod did not affect body or white adipose tissue (WAT) masses, although SDs decreased carcass lipid in males. Free-water deprivation decreased body and WAT pad masses, and all carcass components. Collectively, these results suggest that changes in day length or water availability alone do not affect reproductive status in Shaw's jird. PMID- 10386912 TI - Effects of prenatal morphine exposure on rat heterotypical sexual behavior. AB - Prenatal exposure to morphine inhibits ovarian steroid-dependent lordosis behavior in female rats, and enhances certain components of male sexual behavior in male rats. In the present study, the effects of mid to late gestational morphine exposure on male sexual behavior in females and on female sexual behavior in males were examined in adult offspring. Gonadectomized male rats were injected at weekly intervals with 30 or 60 microg estradiol benzoate and 1.0 mg progesterone and tested for female sexual behavior with stimulus males on 2 consecutive weekly tests. Ovariohysterectomized (OVX) females were injected with 500 microg testosterone propionate (TP) daily for 15 days and tested for male sexual behavior with stimulus females on the last day of TP injection and 1 week later, after TP withdrawal. Prenatal morphine exposure increased the expression of male sexual behaviors in female rats, but it did not increase lordosis behavior in male rats. Thus, exposure to morphine during gestation alters male and female sexual behavior in young adult animals. Because prenatal morphine exposure both defeminized and masculinized adult sexual behavior in female rats, it is possible that female brain development is more vulnerable to prenatal insult such as opiate exposure. PMID- 10386913 TI - Behavior of adult and aged mice before and after central injection of interleukin 1beta. AB - The level of locomotor activity, body temperature (T(B)), and feeding for adult (3-5-month old) and aged (22-24-month old) male BALB/c mice was determined and the sensitivity of the two age groups to the anorectic, febrile, and behavioral properties of interleukin-1beta (IL-1beta) in the brain was examined. Baseline locomotor activity and T(B) were markedly lower in aged mice than in adults and the circadian rhythm for both activity and T(B) were disrupted in the aged. Adult and aged mice consumed similar amounts of food during the daytime and nighttime, but aged mice made longer, less frequent visits to the feed cup. To determine if aging affects the responsiveness to central IL-1beta, adult and aged mice were injected intracerebroventricularly with PBS or IL-1beta. Compared to age-matched PBS controls, IL-1beta increased T(B) in both adult and aged mice. The peak deltaT(B) was greater in aged mice than in adults, but because of a lower baseline T(B) in aged mice, peak T(B) after IL-1beta was not different between groups. Locomotor activity of aged mice receiving PBS was about half that of PBS injected adults and was not depressed further by IL-1beta. However, compared to age-matched PBS controls, centrally administered IL-1beta depressed food intake more in aged mice than in adults. These data indicate that even though feeding, locomotor activity, and T(B) are affected by aging, the central component of the inflammatory response mediated by IL-1beta is retained. PMID- 10386914 TI - Palatability affects satiation but not satiety. AB - The present study was designed to investigate the effect of the pleasantness of a food on satiation (meal termination) and satiety. It was also studied whether or not the subsequent availability of other attractive foods affected the effect of palatability on intake. In a within-subjects repeated-measures design, 35 (26 female and 9 male) young healthy nonrestrained subjects consumed at lunchtime a preload consisting of tomato soup, and a buffet/test meal consisting of many attractive food items. Three factors were manipulated. The palatability of the preload was manipulated by varying the citric acid concentration of the soup at three levels: 0 (pleasant), 7.5 (less pleasant), and 15 (unpleasant) g citric acid/kg soup. Intake of the soup was either ad lib (for investigation of satiation), or standardized (350 g for women, and 500 g for men; for investigation of satiety). The third factor was the availability of other foods, manipulated by the amount of time between start of preload and start of the test meal (intermeal interval = IMI), which was set at two levels: 15 and 90 min. Subjects rated hunger and satiety feelings, before the preload, and in between preload and test meal. The results showed that the ad lib intakes of the less pleasant and unpleasant soups were about 65 and 40% of the intake of the pleasant soup. Subjects ingested about 20% more soup when the subjects had to wait for the test meal about 90 min, compared to the 15 min IMI condition. The availability of other foods had no effect on the effect of pleasantness on ad lib intake. There was also no effect of the pleasantness on subsequent satiety: hunger ratings and test meal intake were similar after the three standardized soups. One conclusion is that pleasantness of foods has an effect on satiation but not on subsequent satiety. A second conclusion is that people eat more of a food when they know that they have no access to other foods for a particular amount of time. PMID- 10386915 TI - A demonstration of classical conditioning of the human eyeblink to an olfactory stimulus. AB - While acquisition of the eyeblink conditioned response to a variety of stimuli has been widely studied, it has yet to be established that humans will demonstrate a conditioned response to an olfactory stimulus. In this study we present data to show that humans will demonstrate a classically conditioned eyeblink response to an olfactory stimulus. Ten participants were tested in a delay conditioning procedure with an olfactory stimulus presented in a heated, humidified stream of air via an olfactometer, allowing the precise control over stimulus duration necessary for delay conditioning. Trials on which odor alone was presented were administered to four additional participants. Establishing that humans will demonstrate an eyeblink conditioned response to an olfactory stimulus will allow further exploration of the pathways involved in classical conditioning and associative learning, as well as an analysis of conditioning pathways across sensory modalities. PMID- 10386916 TI - Scent-marking and cortisol response in the small-eared bushbaby (Otolemur garnettii). AB - Among prosimians, some types of scent-marking may serve as displacement activities that reduce physiological arousal in stressful situations. Type and frequency of scent-marking was measured for 22 male small-eared or Garnett's bushbabies (Otolemur garnettii) exposed to a novel open field environment, with and without novel objects. Rates of foot rubbing, chest rubbing, urine washing, flank rubbing, and ano-genital marking were measured. Foot and chest rubbing constituted 92.5% of responses. Type and frequency of scent-marking was compared to the magnitude of the animals' cortisol responses in a separate test of restraint stress. Only foot and chest rubbing were systematically related to cortisol levels. The animals that performed these behaviors more in the novel environment also exhibited lower cortisol responses to restraint stress. These results suggest that bushbabies that characteristically employ behavioral coping strategies have a reduced physiological response to psychological stressors. PMID- 10386917 TI - Generalization between binary odor mixtures and their components in the rat. AB - We have adopted a conditioning paradigm to investigate generalization between odor mixtures and components. Rats were conditioned to find a reward buried in odor-scented cups. The conditioned odor was either a mixture (O1 + O2) or a pure component (O1). Once they learned the task to criterion, they were tested in random sequence for response to that O1, O1 + O2 and to an unrelated odor (O3). Generalization was consistently the strongest from O1 to O1 + O2 or from O1 + O2 to O1. Furthermore. the degree of generalization depended on the odorants used as O1, O2, and O3. This latter finding in a particular indicates that this assay can be used to assess properties of mixtures, which could arise at either peripheral or more central locations. PMID- 10386918 TI - 2-Deoxy-D-glucose and mercaptoacetate induce different patterns of macronutrient ingestion. AB - 2-Deoxy-D-glucose (2DG) and mercaptoacetate (MA) are antimetabolic agents that reduce the metabolism of glucose and fatty acids, respectively, and stimulate feeding. The present study compared the effects of MA and 2DG on macronutrient self-selection. Because 2DG and MA have different metabolic actions and appear to activate different neural pathways, our hypothesis was that 2DG and MA would elicit different patterns of macronutrient selection. The first experiment examined macronutrient selection in response to 2DG, MA, and 0.9% saline in rats maintained on a three-macronutrient self-selection diet consisting of cornstarch, casein, and vegetable oil. Subsequently, one macronutrient source was replaced in each of three similar experiments with Polycose, albumin, or solid vegetable shortening. Finally, 2DG and MA tests were conducted in which only one macronutrient (cornstarch, casein, or oil) was available during the test. Results show that MA and 2DG elicit different macronutrient preferences. 2DG elicits intake of all three macronutrients in the same relative proportion consumed during spontaneous feeding across a number of dietary conditions, suggesting that glucoprivation activates interoceptive signals and neural pathways similar to those involved in normal hunger. MA elicits a selective intake of protein. Conditions in which carbohydrate palatability is enhanced or protein palatability is diminished lead to a relative increase in carbohydrate intake in response to MA. However, MA did not increase the intake of fat. Results suggest that intake of each macronutrient is subject to separate neural or endocrine control, and that these controls are linked to metabolic cues. PMID- 10386920 TI - Uridine phosphorylase inhibitors: chemical modification of benzyloxybenzyl barbituric acid and its effects on urdpase inhibition. AB - 5-(o-Benzyloxy)benzylbarbituric acid (6) and 5-(p-benzyloxy)benzylbarbituric acid (7) were prepared and their inhibitory activities compared to 5-(m-benzyloxy) benzylbarbituric acid (BBB) a known, potent inhibitor of uridine phosphorylase (UrdPase). Compounds 6 and 7 were 18-fold and 51-fold less active, respectively, than BBB in inhibiting UrdPase. These data provide solid evidence that the 5 benzylbarbituric acids possessing meta substituents are the most active inhibitors. In addition, 2-thioBBB (11) was synthesized and it was shown to be as active an inhibitor as BBB. PMID- 10386919 TI - The tachykinin NK-1 receptor antagonist, RP-67580, infused into the ventral tegmental area prevents stress-induced analgesia in the formalin test. AB - Substance P (SP) receptors in the ventral tegmental area (VTA) play a critical role in mediating the stress-induced activation of midbrain ascending dopamine (DA) neurons. Interestingly, SP acting in the VTA induces analgesia in the formalin test for tonic pain. Because exposure to stress inhibits pain in this test, we speculated that SP receptors in the VTA might mediate stress-induced analgesia. The present study explored this idea by examining the effect of blocking tachykinin NK-1 receptors in the VTA on footshock stress-induced analgesia in the formalin test. Intra-VTA infusions of the novel tachykinin NK-1 receptor antagonist, RP-67580, prevented this response. This finding suggests that exposure to stress inhibits tonic pain through the release of endogenous SP in the VTA. PMID- 10386921 TI - Nonpeptidic HIV protease inhibitors: 6-alkyl-5,6-dihydropyran-2-ones possessing achiral 3-(4-amino/carboxamide-2-t-butyl,5-methylphenyl thio) moiety: antiviral activities and pharmacokinetic properties. AB - Dihydropyran-2-ones possessing amino and carboxamide functionalities on 3-SPh (2 tert-butyl, 5-methyl) ring were synthesized and evaluated for their antiviral activities. Both the enantiomers of inhibitor 15 were synthesized. The in vitro resistance profile, inhibitory activities against cytochrome P450 isozymes and pharmacokinetic properties of inhibitor 15S will be discussed. PMID- 10386922 TI - A new monoclonal anti-idiotypic catalytic antibody with a CPA-like activity. AB - IIF9D8, a new monoclonal anti-idiotypic catalytic antibody with a CPA esterase like activity was elicited by ID11D7, the monoclonal competitive inhibitory antibody to CPA. The hydrolysis of hippuryl-DL-phenyllactic acid by McAb IIF9D8 follows the Michaelis-Menten kinetics. The Km value and kcat are 0.036 M and 0.598 min(-1), respectively, and the rate acceleration (kcat/kuncat) is 30500. Compared with the previous McAb 32C3 induced by polyclonal antibodies to CPA, McAb IIF9D8 shows higher catalytic efficiency The catalytic antibodies with the catalytic properties similar to natural enzymes could be obtained by this approach. PMID- 10386923 TI - Synthesis and evaluation of glucocerebrosidase inhibitory activity of anhydro deoxyinositols from (+)-epi- and (-)-vibo-quercitols. AB - Twelve 1,2- and 2,3-anhydro-1,2,3,4,5-cyclohexanepentols were synthesized from (+)-epi- and (-)-vibo-quercitols, readily available by bioconversion of myo inositol, and assayed for inhibitory activity against glucocerebrosidase (mouse liver). Among them 1L-1,2-anhydro-1,2,4/3,5-cyclohexanepentol, the 3-deoxy derivative of the irreversible inhibitor conduritol B epoxide (CBE), has been demonstrated to be a highly potent and specific inhibitor, almost comparable to the parent compound. PMID- 10386924 TI - Synthesis and biological evaluation of alpha-mannosidase inhibitory activity of three deoxy derivatives of mannostatin A. AB - Three deoxy derivatives of alpha-mannosidase inhibitor mannostatin A have been synthesized and their inhibitors activity for Jack beans alpha-mannosidase evaluated in order to elucidate roles of each hydroxyl groups of the inhibitor The 1- and 2-deoxy derivatives have preserved inhibitory potentials although they lowered the activity one-hundred fold compared to the parent, but the 3-deoxy derivative lost activity. PMID- 10386925 TI - Synthesis of phosphonate 3-phthalidyl esters as prodrugs for potential intracellular delivery of phosphonates. AB - A new prodrug approach for intracellular delivery of phosphonates was developed via the synthesis of 3-phthalidyl esters of 1-naphthalenemethylphosphonate. This approach is advantageous over the traditional acyloxymethyl phosphonate prodrugs, because these prodrugs do not generate formaldehyde and have improved plasma half lives. PMID- 10386926 TI - Design of the first highly potent and selective aminopeptidase N (EC 3.4.11.2) inhibitor. AB - A series of phosphinic compounds mimicking the transition state of substrates hydrolysed by aminopeptidase N (EC 3.4.11.2) were synthesized. These new compounds have potent inhibitory activities with Ki values in the nanomolar range. These derivatives behave as the most potent APN inhibitors designed to date. PMID- 10386927 TI - Liquid phase parallel synthesis of guanidines. AB - Combinatorial synthesis of N,N'-di(Boc)-Protected guanidines containing piperazine and pyrrolidine scaffolds has been developed. We initiate a preliminary study on the reactivity of several guanylating reagents with soluble polymer-bound diamines in liquid phase. Guanidines are liberated from the polymer support under mild conditions in high yields and high purity by simple precipitation and washings. This combinatorial liquid-phase methodology proves to be a useful tool for constructing guanidine libraries containing diamine scaffolds. PMID- 10386928 TI - Isolation and synthesis of a novel immunosuppressive 17alpha-substituted dammarane from the flour of the Palmyrah palm (Borassus flabellifer). AB - The novel triterpene 1 with a dammarane skeleton and a hitherto unknown 17alpha substitution pattern has been isolated from the Palmyrah palm in low yield and prepared by synthesis in larger quantities. 1 was shown to be an extremely potent immunosuppressant in vitro (MLR; IC50 = 10 ng/ml) and in vivo (DTH; ED50 = 0.01 mg/kg p.o.). A glucocorticoid like activity is excluded. PMID- 10386929 TI - The sulfonimidamide as a novel transition state analog for aspartic acid and metallo proteases. AB - We have developed a novel strategy for the preparation of tetrahedral transition state analogs for aspartic acid and metallo-proteases based upon the sulfonimidamide functional group. Our best alpha-des-amino dipeptide analog binds at least 100-fold tighter than the corresponding ground state structure (i.e., amide). A previously unpublished five-membered cyclic sulfonimidamide was also synthesized. PMID- 10386930 TI - Isolation and characterization of an active-site peptide from a sterol methyl transferase with a mechanism-based inhibitor. AB - Chemical affinity labeling of pure sterol methyl transferase (SMT) from Saccharomyces cerevisiae using the mechanism-based irreversible inhibitor, [3 3H]26,27-dehydrozymosterol, inhibited the SMT with an apparent Ki of 1.1 microM and k(inact) of 1.52 min(-1). The protein-inhibitor adduct was subjected to cleavage with trypsin and the resulting covalently modified peptide was analyzed by Edman sequencing from the N-terminus. The radiochemically labeled ca. 5.0 kDa peptide fragment of the cleavage mixture was shown to be contiguous through 17 residues to a segment that includes a highly conserved hydrophobic motif (Region I, stretching between T78 and F91) characteristic of SMT enzymes. The results confirm that Region I is the sterol binding/active site. PMID- 10386931 TI - Liquid phase synthesis of arylamines and its application to the benzimidazolone via nucleophilic aryl substitution. AB - A method for soluble, inexpensive polymer-supported synthesis of aryl amines and benzimidazolone on the basis of nucleophilic aryl substitution (S(N)Ar) is described. This method involves a direct coupling reaction between resin bound aryl fluoride and amines at ambient temperature. The products are isolated in quantitative yields and excellent purity by simple precipitation and washing. This liquid phase method proves to be a useful tool for constructing combinatorial arylamine and benzimidazolone libraries. PMID- 10386932 TI - Synthesis and evaluation of hapalosin and analogs as MDR-reversing agents. AB - The marine natural product hapalosin and 22 analogs, which incorporated systematic substituent deletions or variations, were prepared. These compounds were evaluated in a cell-based assay for both MDR-reversing activity and general cytotoxicity. Some substituent modifications resulted in lower cytotoxicities, but most structural changes were either detrimental to or did not seriously alter the MDR-reversing activity. PMID- 10386934 TI - Dual-acting agents with alpha1-adrenoceptor antagonistic and steroid 5alpha reductase inhibitory activities. Synthesis and evaluation of arylpiperazine derivatives. AB - A series of arylpiperazine derivatives were prepared and evaluated for their alpha1-adrenoceptor antagonistic activities and 5alpha-reductase inhibitory activities. SAR study led to the identification of the potent dual-acting compound 2f, which had a pA2 value of 7.5 for alpha1-adrenoceptor antagonism and an IC50 value of 1.5 nM for 5alpha-reductase inhibition. PMID- 10386933 TI - Potential anxiolytic agents. 3. Novel A-ring modified pyrido[1,2 a]benzimidazoles. AB - A variety of pyrido[1,2-a]benzimidazoles (PBIs) modified on the A-ring were prepared and evaluated for affinity to the benzodiazepine binding site on the GABA-A receptor and in animal models predictive of anxiolytic activity in humans. A-ring benzo-fused derivative 7 exhibited potent activity, as did the 6- and 7 pyrido compounds 3 and 4. PMID- 10386935 TI - Preparation of crystalline p-nitrobenzyl 2-formyl carbapenems by oxidative cleavage. AB - Crystalline 1beta-methyl-2-formyl carbapenem pNB esters were prepared by osmium mediated oxidative cleavage of the corresponding 2-vinyl derivatives. Reduction of the 2-formyl compounds gave the corresponding 2-hydroxymethyl derivatives, which are key intermediates for the anti-MRS carbapenem candidate (1). PMID- 10386936 TI - 5-Substituted pyrimidine 1,5-anhydrohexitols: conformational analysis and interaction with viral thymidine kinase. AB - Conformational analysis of anhydrohexitol nucleosides using a combination of experimental (X-ray crystallography) and computational methods indicates that those antiviral compounds occur in an equilibrium between two forms, one conformation being adopted in solid phase and in solution, the other found when the nucleosides are in complex with HSV-1 thymidine kinase. The conformational change induced by the enzyme has been investigated. PMID- 10386937 TI - Synthesis of polyamine derivatives for the preparation of affinity chromatography columns for the search of new Trypanosoma cruzi targets. AB - The most potent trypanocidal compound of a series of symmetrically substituted 1,4-bis(3-aminopropylpiperazines) which displayed an IC50 value of 5 microM on Trypanosoma cruzi trypomastigotes, was inactive on trypanothione reductase. Two derivatives 6 and 12 of this compound, one symmetrical and one dissymmetrical, were synthesized via a reductive amination reaction, to prepare affinity chromatography columns, which allowed us to isolate three parasitic proteins. Among these, the major ligand 6- and 12-binding protein having an apparent molecular weight of 52 kDa has been identified as the thiol-disulfide oxido reductase Tc52, previously characterized in Trypanosoma cruzi. PMID- 10386938 TI - Structure-activity relationships of substituted 5H-thiazolo[3,2-a]pyrimidines as group 2 metabotropic glutamate receptor antagonists. AB - A series of 5H-thiazolo[3,2-a]pyrimidine derivatives 1 was studied with respect to the inhibition of 1S,3R-ACPD (10 microM)-stimulated GTP gamma35S binding on rat mGlu2 receptor transfected cell membranes. The influence of substituents at position 6 and 7 as well as the substitution pattern of the two phenyl-rings in position 2 and 5 on the activity is discussed. PMID- 10386939 TI - Lipase-catalyzed protection of the hydroxy groups of the nucleosides inosine and 2'-deoxyinosine: a new chemoenzymatic synthesis of the antiviral drug 2',3' dideoxyinosine. AB - The selective acylation of the hydroxy groups of the nucleosides inosine 1a and 2'-deoxyinosine 1b has been achieved in the presence of Candida antarctica and Pseudomonas sp. lipases in organic solvents; starting from the 5'-acetyl derivative of 2'-deoxyinosine, compound 5a, an efficient chemoenzymatic synthesis of the antiviral drug 2',3'-dideoxyinosine 1c has been achieved. PMID- 10386941 TI - An efficient stereoselective synthesis of [3S(1S,9S)]-3-[[[9-(benzoylamino) octahydro-6,10-dioxo-6H-pyridazino-(1,2-a)(1,2)-diazepin-1-yl]-carbonyl ]amino]-4 oxobutanoic acid, an interleukin converting enzyme (ICE) inhibitor. AB - The title compound 1 is a potent interleukin-1beta-converting enzyme (ICE) inhibitor. Recently, an efficient chiral synthesis of compound 1 has been accomplished in our labs. The overall yield of this 18-step stereoselective synthesis was 9.8%. PMID- 10386940 TI - Novel derivatives of 3-(dipropylamino)chroman. Interactions with 5-HT1A and D2A receptors. AB - Novel 8-aryl and 8-aroyl substituted derivatives of 3-(dipropylamino)chroman are described. The compounds have been prepared by a palladium catalyzed reaction of iodoarenes and a stannylated derivative of [eta6-3 (dipropylamino)chroman]Cr(CO)3. Several of the compounds have high affinity for 5 HT1A receptors whereas the affinity for D2A receptors is lower, the 8-arylated derivatives being slightly more potent than the 8-aroylated analogues. PMID- 10386942 TI - Rational design of N-[2-(2,5-dimethoxyphenylethyl)]-N'-[2-(5-bromopyridyl)] thiourea (HI-236) as a potent non-nucleoside inhibitor of drug-resistant human immunodeficiency virus. AB - The novel thiourea compound N-[2-(2,5-dimethoxyphenylethyl)]-N'-[2-(5 bromopyridyl)]-thi ourea (HI-236) targeting the non-nucleoside inhibitor (NNI) binding pocket of HIV-1 reverse transcriptase (RT) was rationally designed using a computer model of the NNI binding pocket. The NNI binding pocket model takes into consideration changes in binding pocket size, shape, and changes in residue character that result from clinically-observed NNI resistance-associated mutations of HIV RT. RT assays revealed that HI-236 was not only more potent than trovirdine, MKC-442, and AZT against the drug-sensitive HIV-1 strain HTLV(IIIB), it was also 50-100 times more effective than delavirdine or nevirapine and twice as effective as our recently reported lead compound N-[2-(2-fluorophenethyl)]-N' [2-(5-bromopyridyl)]-thiourea (HI-240) against the NNI-resistant Y181C mutant HIV 1 strain A17. Most importantly, HI-236 was highly effective against the multidrug resistant HIV-1 strain RT-MDR with multiple mutations involving the RT residues 74V, 41L, 106A, and 215Y. The activity of HI-236 against RT-MDR was superior to that of other anti-HIV agents tested, which are listed in the following order: HI 236 (IC50: 5 nM) > HI-240 (IC50: 6 nM) > trovirdine (IC50: 20 nM) > AZT (IC50: 150 nM) > MKC-442 (IC50: 300 nM) > delavirdine (IC50: 400 nM) > nevirapine (IC50: 5 microM). PMID- 10386943 TI - Synthesis of 25-aminosterols, new antifungal agents. AB - 25-aminolanostenol 1 and 25-aminocholesterol 2 were hemisynthesized from natural sterols and tested in vitro against Candida albicans. The biological activity of compound 1 was rather weak, whereas 2 exhibited in vitro antifungal activity with MIC value of 4 microM. PMID- 10386944 TI - N-substituted 4-(5-indolyl)benzoic acids. Synthesis and evaluation of steroid 5alpha-reductase type I and II inhibitory activity. AB - The synthesis of N-alkyl and N-arylalkyl substituted 4-(5-indolyl)benzoic acid derivatives as inhibitors of steroid 5alpha-reductases is described. For the human type II isozyme a benzyl substituent (IC50 6.20 microM) and for the human type I isozyme a cyclohexanemethyl substituent (IC50 2.10 microM) on the indole nitrogen proved to be most efficacious, thus providing interesting leads for the development of drugs for the treatment of benign prostatic hyperplasia (BPH). PMID- 10386945 TI - Design of scytalone dehydratase inhibitors as rice blast fungicides: (N phenoxypropyl)-carboxamides. AB - Insights gained from a crystal structure of scytalone dehydratase led to the design of carboxamide inhibitors with a phenoxypropyl group substituted on the nitrogen atom Potent enzyme inhibitors were synthesized around this motif, the best of which provided excellent control of rice blast disease in greenhouse assays and outdoor field trials. PMID- 10386946 TI - Design of scytalone dehydratase inhibitors as rice blast fungicides: derivatives of norephedrine. AB - Five X-ray crystal structures of scytalone dehydratase complexed with different inhibitors have delineated conformationally flexible regions of the binding pocket. This information was used for the design and synthesis of a norephedrine derived cyanoacetamide class of inhibitors leading to potent fungicides. PMID- 10386947 TI - Structure-activity relationship for a series of 2-substituted 1,2,3,4-tetrahydro 9H-pyrido[3,4-b]indoles: potent subtype-selective inhibitors of N-methyl-D aspartate (NMDA) receptors. AB - A series of 2-substituted 1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indoles was synthesized as potential antagonists for the NR1A/2B subtype of N-methyl-D aspartate (NMDA) receptors. Assayed by electrical recording under steady-state conditions, 7-hydroxy-2-(4-phenylbutyl)- 1,2,3,4-tetrahydropyrido-[3,4-b]indole (30) was the most potent compound in the series having an IC50 value of 50 nM at the NR1A/2B receptors. PMID- 10386948 TI - Amino-substituted thalidomide analogs: potent inhibitors of TNF-alpha production. AB - Thalidomide, (1), is a known inhibitor of TNF-alpha release in LPS stimulated human PBMC. Herein we describe the TNF-alpha inhibitory activity of amino substituted analogs of thalidomide (1) and its isoindolin-1-one analog, EM-12 (2). The 4-amino substituted analogs were found to be potent inhibitors of TNF alpha release in LPS stimulated human PBMC. PMID- 10386949 TI - Glycolipid-enriched caveolae and caveolae-like domains in the nervous system. AB - Recent years have been characterized by a booming interest in research on caveolae and caveolae-like membrane domains. The interest in this subject grew further, when their involvement in fundamental membrane-associated events, such as signal transmission and lipid/protein sorting, was postulated. Substantial progress has been reached in understanding the biological role of membrane domains in eukaryotic cells. The neuron, however, which perhaps represents one of the greatest challenges to research on membrane traffic and function, has only been partially investigated. The purpose of the present review is to survey this issue in the nervous system. We confine ourselves to the presence of membrane domains in the nervous system and discuss this in the context of three facts: first, glycolipids are peculiarly enriched in both caveolae and caveolae-like domains and are particularly abundant in the nervous system; second, the neuron is characterized by a basic dual polarity, similar in this respect to other polarized cells, where the role of glycolipid-enriched domains for lipid/protein sorting has been better ascertained; and third, neurons evolved from, and are related to, simpler eukaryotic cells, allowing us to find analogies with more investigated nonneuronal cells. PMID- 10386950 TI - Molecular cloning of testican-2: defining a novel calcium-binding proteoglycan family expressed in brain. AB - We have screened a human cDNA library using an expressed sequence tag related to the BM-40/secreted protein, acidic and rich in cysteine (SPARC)/osteonectin family of proteins and isolated a novel cDNA. It encodes a protein precursor of 424 amino acids that consists of a signal peptide, a follistatin-like domain, a Ca2+-binding domain, a thyroglobulin-like domain, and a C-terminal region with two putative glycosaminoglycan attachment sites. The protein is homologous to testican-1 and was termed testican-2. Testican-1 is a proteoglycan originally isolated from human seminal plasma that is also expressed in brain. Northern blot hybridization of testican-2 showed a 6.1-kb mRNA expressed mainly in CNS but also found in lung and testis. A widespread expression in multiple neuronal cell types in olfactory bulb, cerebral cortex, thalamus, hippocampus, cerebellum, and medulla was detected by in situ hybridization. A recombinant fragment consisting of the Ca2+-binding EF-hand domain and the thyroglobulin-like domain of testican 2 showed a reversible Ca2+-dependent conformational change in circular dichroism studies. Testican-1 and -2 form a novel Ca2+-binding proteoglycan family built of modular domains with the potential to participate in diverse steps of neurogenesis. PMID- 10386951 TI - A PC12 variant lacking regulated secretory organelles: aberrant protein targeting and evidence for a factor inhibiting neuroendocrine gene expression. AB - A variant of the PC12 pheochromocytoma cell line (termed A35C) has been isolated that lacks regulated secretory organelles and several constituent proteins. Northern and Southern blot analyses suggested a block at the transcriptional level. The proprotein-converting enzyme carboxypeptidase H was synthesised in the A35C cell line but was secreted by the constitutive pathway. Transient transfection of A35C cells with cDNAs encoding the regulated secretory proteins dopamine beta-hydroxylase and synaptotagmin I resulted in distinct patterns of mistargeting of these proteins. It is surprising that hybrid cells created by fusing normal PC12 cells with A35C cells exhibited the variant phenotype, suggesting that A35C cells express an inhibitory factor that represses neuroendocrine-specific gene expression. PMID- 10386952 TI - Processing of wild-type and mutant familial Alzheimer's disease-associated presenilin-1 in cultured neurons. AB - Mutations of presenilin (PS)-1, an endoplasmic reticulum/Golgi transmembrane protein, have been associated with early-onset familial Alzheimer's disease (FAD). In mammalian brain, PS1 exists primarily as its processed fragments; however, the role of this cleavage event in PS1 function remains unclear. Although some investigators have shown that mutant PS1 processing is unaltered (with the exception of PS1-deltaE9, which lacks the cleavage site) in stably transfected cells and PS1-FAD transgenic mice, other investigators have reported altered FAD mutant PS1 and PS2 protein processing in transiently transfected cells and human FAD patients. The present study uses recombinant replication defective adenoviral vectors to transiently express wild-type (WT) or mutant PS1 in various cells, including primary cultured hippocampal neurons. We show that in contrast to PS1-WT, overexpression of mutant PS1 results in an increased ratio of mutant holoprotein to endoproteolytic products that is dependent on cell type and differentiation state. In addition, mutant PS1 overexpression leads to an increase in caspase-type protease derived fragments above that seen with PS1-WT overexpression. Furthermore, overexpression of at least one mutant significantly alters the processing of coexpressed PS1-WT, suggesting that mutant PS1 may affect PS1-WT function. These findings suggest that a defect in PS1 holoprotein stability may be a general defect seen in cells expressing mutant PS1, especially neuronal cells, and may play a critical role in the pathogenesis of FAD. PMID- 10386953 TI - Brain-derived neurotrophic factor stimulates interactions of Shp2 with phosphatidylinositol 3-kinase and Grb2 in cultured cerebral cortical neurons. AB - Shp2, a protein tyrosine phosphatase possessing SH2 domains, is utilized in the intracellular signaling of various growth factors. Shp2 is highly expressed in the CNS. Brain-derived neurotrophic factor (BDNF), a member of the neurotrophin family, which also shows high levels of expression in the CNS, exerts neurotrophic and neuromodulatory effects in CNS neurons. We examined how BDNF utilizes Shp2 in its signaling pathway in cultured cerebral cortical neurons. We found that BDNF stimulated coprecipitation of several tyrosine-phosphorylated proteins with anti-Shp2 antibody and that Grb2 and phosphatidylinositol 3-kinase (PI3-K) were coprecipitated with anti-Shp2 antibody in response to BDNF. In addition, both anti-Grb2 and anti-PI3-K antibodies coprecipitated Shp2 in response to BDNF. The BDNF-stimulated coprecipitation of the tyrosine phosphorylated proteins, Grb2, and PI3-K with anti-Shp2 antibody was completely inhibited by K252a, an inhibitor of TrkB receptor tyrosine kinase. This BDNF stimulated Shp2 signaling was markedly sustained as well as BDNF-induced phosphorylation of TrkB and mitogen-activated protein kinases. In PC12 cells stably expressing TrkB, both BDNF and nerve growth factor stimulated Shp2 signaling similarly to that by BDNF in cultured cortical neurons. These results indicated that Shp2 shows cross-talk with various signaling molecules including Grb2 and PI3-K in BDNF-induced signaling and that Shp2 may be involved in the regulation of various actions of BDNF in CNS neurons. PMID- 10386954 TI - Cyclic AMP regulates the expression of neurokinin1 receptors by neonatal rat spinal neurons in culture. AB - Neurokinin1 (NK1) receptors are up-regulated in the spinal cord during peripheral inflammation, but the biochemical mediators regulating this change have not been resolved. The promoter region of the gene encoding the NK1 receptor contains a cyclic AMP (cAMP)-responsive element. Therefore, we used primary cultures of neonatal rat spinal cord to test whether increasing intracellular cAMP can increase expression of NK1 receptors. Treatment with dibutyryl-cAMP (dbcAMP) resulted in a time-dependent increase in 125I-Bolton-Hunter-substance P (BHSP) binding in the cultures; treatment with dibutyryl-cyclic GMP did not. Treatment with forskolin plus 3-isobutyl-1-methylxanthine mimicked the increase in binding, providing further evidence for the involvement of cAMP in this effect. Scatchard analyses indicated that the increase in BHSP binding was due to an increase in binding capacity. The cAMP-induced increase in BHSP binding was preceded by an increase in levels of mRNA for NK1 receptor and was attenuated by pretreatment with cycloheximide. These data indicate that the cAMP-induced increase in binding was due to increased synthesis of NK1 receptors. Comparison of substance P (SP) induced production of inositol phosphates between cultures pretreated with dbcAMP and controls suggested that increased expression of NK1 receptors did not result in increased generation of second messenger by NK1 receptor activation. Together, these data indicate that a persistent increase in intracellular cAMP increases expression of NK1 receptors. Because NK1 receptor activation contributes to increased excitability of spinal neurons, the increased expression of NK1 receptors may be important in maintaining responsiveness of spinal neurons to SP in central mechanisms underlying hyperalgesia. PMID- 10386955 TI - Alpha1-antichymotrypsin-like proteins I and II purified from bovine adrenal medulla are enriched in chromaffin granules and inhibit the proenkephalin processing enzyme "prohormone thiol protease". AB - Proteolytic processing of inactive proenkephalin and proneuropeptides is essential for the production of biologically active enkephalins and many neuropeptides. The incomplete processing of proenkephalin in adrenal medulla suggests that endogenous protease inhibitors may inhibit proenkephalin processing enzymes. This study demonstrates the isolation and characterization of two isoforms of adrenal medullary alpha1-antichymotrypsin (ACT), referred to as ACT like proteins I and II, which are colocalized with enkephalin in chromaffin granules and which inhibit the proenkephalin processing enzyme known as prohormone thiol protease (PTP). Subcellular fractionation demonstrated enrichment of 56- and 60-kDa ACT-like proteins I and II, respectively, to enkephalin-containing chromaffin granules (secretory vesicles). Immunofluorescence cytochemistry of chromaffin cells indicated a discrete, punctate pattern of ACT immunostaining that resembles that of [Met]enkephalin that is stored in secretory vesicles. Chromatography of adrenal medullary extracts through DEAE-Sepharose and chromatofocusing resulted in the separation of ACT-like proteins I and II that possess different isoelectric points of 5.5 and 4.0, respectively. The 56-kDa ACT-like protein I was purified to apparent homogeneity by Sephacryl S200 chromatography; the 60-kDa ACT-like protein II was isolated by butyl-Sepharose, Sephacryl S200, and concanavalin A-Sepharose columns. The proenkephalin processing enzyme PTP was potently inhibited by ACT like protein I, with a K(i,app) of 35 nM, but ACT-like protein II was less effective. ACT-like proteins I and II had little effect on chymotrypsin. These results demonstrate the biochemical identification of two secretory vesicle ACT like proteins that differentially inhibit PTP. The colocalization of the ACT-like proteins and PTP within chromaffin granules indicates that they could interact in vivo. Results from this study suggest that these ACT-like proteins may be considered as candidate inhibitors of PTP, which could provide a mechanism for limited proenkephalin processing in adrenal medulla. PMID- 10386956 TI - Differential effects of glial cell line-derived neurotrophic factor and neurturin on developing and adult substantia nigra dopaminergic neurons. AB - Neurturin (NTN) and glial cell line-derived neurotrophic factor (GDNF), two members of the GDNF family of growth factors, exert very similar biological activities in different systems, including the substantia nigra. Our goal in the present work was to compare their function and define whether nonoverlapping biological activities on midbrain dopaminergic neurons exist. We first found that NTN and GDNF are differentially regulated during postnatal development. NTN mRNA progressively decreased in the ventral mesencephalon and progressively increased in the striatum, coincident with a decrease in GDNF mRNA expression. This finding suggested distinct physiological roles for each factor in the nigrostriatal system. We therefore examined their function in ventral mesencephalon cultures and found that NTN promoted survival comparable with GDNF, but only GDNF induced sprouting and hypertrophy of developing dopaminergic neurons. We subsequently examined the ability of NTN to prevent the 6-hydroxydopamine-induced degeneration of adult dopaminergic neurons in vivo. Fibroblasts genetically engineered to deliver high levels of GDNF or NTN were grafted supranigrally. NTN was found to be as potent as GDNF at preventing the death of nigral dopaminergic neurons, but only GDNF induced tyrosine hydroxylase staining, sprouting, or hypertrophy of dopaminergic neurons. In conclusion, our results show selective survival promoting effects of NTN over wider survival, neuritogenic, and hypertrophic effects of GDNF on dopaminergic neurons in vitro and in vivo. Such differences are likely to underlie unique roles for each factor in postnatal development and may ultimately be exploited in the treatment of Parkinson's disease. PMID- 10386957 TI - CDK-5-mediated neurofilament phosphorylation in SHSY5Y human neuroblastoma cells. AB - Cyclin-dependent kinase-5 (CDK-5) has been shown to play important roles in neuronal development and neurogenesis. In vitro studies indicate a role of CDK-5 in phosphorylation of neurofilaments (NFs). In this study, we have chosen the human neuroblastoma cell line SHSY5Y as a model system to study the in vivo phosphorylation of NF proteins by CDK-5. Upon differentiation of SHSY5Y cells with retinoic acid, we found that the phosphorylation of high molecular mass (NF H) and medium molecular mass (NF-M) NFs increased, whereas the CDK-5 protein level and kinase activity were unaffected. The role of CDK-5 in the phosphorylation of cytoskeletal proteins was studied by using antisense oligonucleotides (ONs) to inhibit the expression of the CDK-5 gene. We found that inhibition of CDK-5 levels by antisense ON treatment resulted in a decrease in phosphorylation of NF-H that correlated with a decline in neurite outgrowth. These results demonstrate that CDK-5 is a major proline-directed kinase phosphorylating the human NF-H tail domain. PMID- 10386958 TI - Evidence for phosphatidylinositol 4-kinase and actin involvement in the regulation of 125I-beta-nerve growth factor retrograde axonal transport. AB - The signaling events regulating the retrograde axonal transport of neurotrophins are poorly understood, but a role for phosphatidylinositol kinases has been proposed. In this study, we used phenylarsine oxide (PAO) to examine the participation of phosphatidylinositol 4-kinases in nerve growth factor (NGF) retrograde axonal transport within sympathetic and sensory neurons. The retrograde transport of 125I-labeled betaNGF was inhibited by PAO (0.5-2 nmol/eye), and this effect was diminished by dilution. Coinjection of 2,3 dimercaptopropanol with PAO reduced its ability to inhibit 125I-betaNGF retrograde transport. PAO (20 nM to 200 microM) also inhibited NGF-dependent survival of both sympathetic and sensory neuronal populations. F-actin staining in sympathetic and sensory neuronal growth cones was disrupted by PAO at 10 and 2 nM, respectively, and occurred within 5 min of exposure to the drug. The actin inhibitor latrunculin A also rapidly affected F-actin staining in vitro and reduced 125I-betaNGF retrograde axonal transport in vivo to the same extent as PAO. These results suggest that both phosphatidylinositol 4-kinase isoforms and the actin cytoskeleton play significant roles in the regulation of 125I-betaNGF retrograde axonal transport in vivo. PMID- 10386959 TI - Leukemia inhibitory factor is an autocrine survival factor for Schwann cells. AB - Schwann cells play a major role in promoting nerve survival and regeneration after injury. Their activities include providing neurotrophic factors and increasing the production of extracellular matrix components and cell surface adhesion molecules to promote axon regeneration. Following nerve transection, leukemia inhibitory factor (LIF) is up-regulated by Schwann cells at the injury site. LIF receptors are also up-regulated at the nerve injury site, but their cellular localization and function have not been fully characterized. We demonstrate that Schwann cells express mRNAs for LIF and the LIF receptor components LIF receptor subunit beta and glycoprotein 130 in vitro. We also show that although LIF is not required for the genesis of Schwann cells, it can potentiate the survival of differentiated Schwann cells in the context of neuregulin support. Not only does exogenous LIF promote survival under these conditions, but addition of the soluble LIF receptor (LIF binding protein) and anti-LIF antibodies significantly reduced cell survival, suggesting that LIF exerts autocrine effects. These results suggest that Schwann cell survival following nerve injury is potentially modulated by LIF. PMID- 10386960 TI - Effect of dibutyryl cyclic AMP on the kinetics of myo-inositol transport in cultured astrocytes. AB - Dibutyryl cyclic AMP (dBcAMP) is known to induce maturation and differentiation in astrocytes. As myo-inositol is an important osmoregulator in astrocytes, we examined the effects of maturation and biochemical differentiation on the kinetic properties of myo-inositol transport. Treatment of astrocytes with dBcAMP significantly decreased the Vmax of myo-inositol uptake, but the effect on Km was not significant. The myo-inositol content of astrocytes was significantly decreased in cells treated for 5 days with dBcAMP as compared with untreated controls. Maximum suppression of myo-inositol uptake occurred 7 days after exposure of astrocytes to dBcAMP; this was gradually reversible when dBcAMP was removed from the medium. After exposure to hypertonic medium for 6 h, mRNA expression of the myo-inositol co-transporter was diminished by approximately 36% in astrocytes treated with dBcAMP as compared with untreated cells. It appears that myo-inositol transporters in astrocytes treated with dBcAMP are either decreased in number or inactivated during maturation and differentiation, suggesting that the stage of differentiation and biochemical maturation of astrocytes is an important factor in osmoregulation. PMID- 10386961 TI - Glial cells mediate toxicity in glutathione-depleted mesencephalic cultures. AB - We have examined the role of glial cells in the toxicity that results from inhibition of reduced glutathione (GSH) synthesis by L-buthionine sulfoximine (BSO) in mesencephalic cell cultures. We show that GSH depletion, to levels that cause total cell loss in cultures containing neurons and glial cells, has no effect on cell viability in enriched neuronal cultures. An increase in the plating cell density sensitizes glia-containing cultures to GSH depletion-induced toxicity. This suggests that cell death in this model is the consequence of events that are induced by GSH depletion and are mediated by glial cells. The antioxidant ascorbic acid and the lipoxygenase (LOX) inhibitor nordihydroguaiaretic acid (1-10 microM) provide full protection from BSO toxicity, indicating that arachidonic acid metabolism through the LOX pathway and the generation of reactive oxygen species play a role in the loss of cell viability. In contrast, inhibition of nitric oxide (NO) synthase affords only partial protection from BSO toxicity, suggesting that increased NO production cannot entirely account for cell death in this model. Our data provide evidence that GSH depletion in the presence of glial cells leads to neuronal degeneration that can be prevented by inhibition of LOX. This may have relevance to the pathogenesis of Parkinson's disease, where glial activation and depletion of GSH have been found in the substantia nigra pars compacta. PMID- 10386962 TI - Analysis of cis-acting sequences from the myelin oligodendrocyte glycoprotein promoter. AB - Myelin oligodendrocyte glycoprotein (MOG), a minor component of the myelin sheath, appears to be implicated in the late events of CNS myelinogenesis. To investigate the transcriptional regulation of MOG, 657 bp of the 5'-flanking sequence of the murine MOG gene, previously shown to induce the highest level of transcription in an oligodendroglial cell line, was analyzed by in vitro footprinting and electrophoretic mobility shift assays. This region contains at least three sites that contact nuclear proteins in vitro. Each region described in this study binds specific nuclear proteins and enhances transcription in the OLN-93 glial cell line. More specifically, a region located at position -93 to 73 bp, which displays 100% homology in mouse and human MOG promoters, presents distinct binding affinities between brain and liver nuclear proteins. The results obtained by supershift assay and site-directed mutagenesis reveal that this region contains an essential positive element (TGACGTGG) related to the cyclic AMP-responsive element CREB-1 and are additional evidence for the involvement of the cyclic AMP transduction pathway in oligodendrocyte development. PMID- 10386963 TI - Cyclic AMP-dependent activation of the proenkephalin gene requires phosphorylation of CREB at serine-133 and a Src-related kinase. AB - The transcription factor CREB [cyclic AMP response element (CRE)-binding protein] is activated by several kinase pathways on phosphorylation of serine-133. Phosphorylation of CREB at serine-133 is required for the induction of target gene expression. The proenkephalin gene is a target of cyclic AMP-dependent agonists like forskolin, and its expression is driven by the enhancer element CRE 2. It has been shown that CREB binds CRE-2 in extracts from striatum and hypothalamus. However, these studies did not show a functional requirement for CREB serine-133 phosphorylation in CRE-2 function. We demonstrate that CREB binds CRE-2 in primary astrocyte cultures and that transcriptional activation of CRE-2 requires CREB phosphorylation at serine-133. In addition, it has recently been shown that, at least in some contexts, CREB phosphorylation is not sufficient to activate target gene expression and that another intracellular signal seems to be required. Therefore, we also sought to determine if another signaling event, in addition to CREB phosphorylation, might be involved in cyclic AMP-mediated induction of the proenkephalin gene. We have found that the inhibition of src related nonreceptor tyrosine kinases blocks forskolin-induced proenkephalin gene expression without having any effect on serine-133-phosphorylated CREB levels and that constitutively activated src kinase can activate the proenkephalin promoter. PMID- 10386964 TI - Neurotrophins differentially regulate the survival and morphological complexity of human CNS model neurons. AB - To determine the effect of neurotrophins on the survival and morphological differentiation of CNS neurons, we examined NT2-N cells, which provide a unique culture model for terminally differentiated and polar human neurons. Here we report the development of conditions for the long-term culture of NT2-N cells in low density and in chemically defined medium. We show that NT2-N cells express rRNAs for TrkA, TrkB, and TrkC tyrosine kinase receptors and the low-affinity nerve growth factor receptor (p75NTR). All members of the nerve growth factor related family of neurotrophic factors promote neuronal survival in long-term cultures with approximately 1 ng/ml for half-maximal survival. At high concentrations (>20 ng/ml), the neurotrophins reversed the survival-promoting effect as judged by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] conversion. In contrast to the uniform effect of all neurotrophins on neuronal survival, brain-derived neurotrophic factor selectively induced an increased dendritic complexity. These results demonstrate that NT2-N cells provide a useful model to analyze the effect of neurotrophins on the survival and morphological differentiation of CNS neurons in vitro. In addition, the data indicate that neuronal survival and the development of morphological complexity are differentially regulated in a multireceptor context. PMID- 10386965 TI - Calcium regulation of cyclic nucleotide signaling in lobster olfactory receptor neurons. AB - An elevated free Ca2+ concentration reduces odor-stimulated production of cyclic AMP (cAMP) in the outer dendritic membranes of lobster olfactory receptor neurons in vitro. This effect can occur within 50 ms of odor stimulation. The effect is concentration-dependent at submicromolar concentrations of free Ca2+. An elevated free Ca2+ concentration also reduces basal and forskolin-stimulated cAMP levels in a concentration-dependent manner, suggesting that Ca2+ is not targeting the activation of the odor receptor/G protein complex. The degradation of synthetic cAMP by phosphodiesterases is not enhanced by an increased free Ca2+ concentration, suggesting that Ca2+ acts by down-regulating the olfactory adenylyl cyclase. Western blot analysis of the lobster olfactory sensilla that contain the outer dendrites reveals a protein in the transduction zone with a molecular mass of approximately 138 kDa that is immunoreactive to an antiserum against adenylyl cyclase type III. Given earlier evidence that Ca2+ potentially enters the receptor cell through odor-activated inositol 1,4,5-trisphosphate gated channels, our results suggest a possible route for cross talk between the cyclic nucleotide and the inositol phospholipid signaling pathways in lobster olfactory receptor neurons. PMID- 10386966 TI - Regulation of tyrosine hydroxylase gene expression by muscarinic agonists in rat adrenal medulla. AB - Tyrosine hydroxylase (TH) gene expression in the adrenal medulla is regulated by numerous stimuli via transsynaptic mechanisms. The adrenal chromaffin cell receptors that mediate this transsynaptic response remain unidentified. In this report we demonstrate that the muscarinic acetylcholine receptor agonist bethanechol stimulates the TH gene transcription rate in both innervated and denervated adrenal glands. Hence, this muscarinic response is not dependent on transsynaptic influences, suggesting that agonist occupation of adrenal chromaffin cell muscarinic receptors is sufficient to activate intracellular signaling pathways that stimulate the TH gene. When bethanechol is administered repeatedly over a 3-h interval (four injections spaced 1 h apart), TH mRNA levels are increased two- to threefold at 6 and 12 h after the initial injection of drug. It is surprising that this induction of TH mRNA does not lead to increases in TH activity or TH protein level. These results are consistent with the hypothesis that both transcriptional and posttranscriptional mechanisms must be regulated to induce TH protein and that muscarinic agonists activate only a subset of these mechanisms. PMID- 10386967 TI - Differential coupling of serotonin 5-HT1A and 5-HT1B receptors to activation of ERK2 and inhibition of adenylyl cyclase in transfected CHO cells. AB - Although the subtypes of serotonin 5-HT1 receptors have distinct structure and pharmacology, it has not been clear if they also exhibit differences in coupling to cellular signals. We have sought to compare directly the coupling of 5-HT1A and 5-HT1B receptors to adenylyl cyclase and to the mitogen-activated protein kinase ERK2 (extracellular signal-regulated kinase-2). We found that 5-HT1B receptors couple better to activation of ERK2 and inhibition of adenylyl cyclase than do 5-HT1A receptors. 5-HT stimulated a maximal fourfold increase in ERK2 activity in nontransfected cells that express endogenous 5-HT1B receptors at a very low density and a maximal 13-fold increase in transfected cells expressing 230 fmol of 5-HT1B receptor/mg of membrane protein. In contrast, activation of 5 HT1A receptors stimulated only a 2.8-fold maximal activation of ERK2 in transfected cells expressing receptors at 300 fmol/mg of membrane protein but did stimulate a 12-fold increase in activity in cells expressing receptors at 3,000 fmol/mg of membrane protein. Similarly, 5-HT1A, but not 5-HT1B, receptors were found to cause significant inhibition of forskolin-stimulated cyclic AMP accumulation only when expressed at high densities. These findings demonstrate that although both 5-HT1A and 5-HT1B receptors have been shown to couple to G proteins of the Gi class, they exhibit differences in coupling to ERK2 and adenylyl cyclase. PMID- 10386968 TI - Protein synthesis blockade differentially affects the degradation of constitutive and nicotinic receptor-induced tyrosine hydroxylase protein level in isolated bovine chromaffin cells. AB - Continuous incubation of bovine adrenal chromaffin cells with the nicotinic receptor agonist 1,1-dimethyl-4-phenylpiperazinium (DMPP) causes a twofold increase in the steady-state level of catalytically active tyrosine hydroxylase (TH) protein by 3-4 days. The present study examined the processes that control the time course of enzyme induction. In cells exposed to DMPP for 36 or 54 h, incorporation of [3H]leucine into TH was increased 1.9- and 2.2-fold, respectively, compared with control (non-DMPP-treated) cells. The increase correlated with a twofold rise in TH mRNA level, indicating the absence of translational control of TH synthesis by DMPP. Also absent was an effect by DMPP on the rate of degradation of TH protein because pulse-chase analysis estimated a half-life for TH of 26 +/- 5 h in DMPP-treated cells, a value that was (a) essentially the same as that estimated in control cells (29 +/- 3 h), (b) within the same range as that estimated by approach to steady state (t(1/2) = 19 +/- 4 h), which measured the decline of TH protein content from the DMPP-induced steady state level back to the basal value during deinduction with the nicotinic antagonist hexamethonium, and (c) consistent with the time course of accumulation of TH protein to a new steady-state level in response to DMPP. However, different rates of degradation for TH protein were observed in control and DMPP-treated cells under conditions in which protein synthesis was blocked. In control cells incubated with 100 microM puromycin or 20 microM cycloheximide for 3 days, the level of catalytically active TH protein failed to decline and exhibited a half life of > or = 250 h. This finding indicated that TH protein was stabilized. TH protein level also failed to decline when cells were incubated for 3 days with a concentration of the transcription inhibitor alpha-amanitin that caused a >90% loss of TH mRNA. Thus, degradation of constitutively expressed TH protein appears to be controlled by processes dependent on ongoing transcription and translation. In contrast, the increased amount of TH induced by DMPP was not stabilized but instead underwent a decline to the basal level following addition of puromycin or cycloheximide. It is important to note, however, that the decline occurred at a slower rate (t(1/2) > or = 45 h) than that measured during deinduction. Taken together, these data suggest that alterations in the rate of degradation of TH protein may play a role in controlling TH level when TH synthesis is blocked but not when TH synthesis is increased, such as during nicotinic receptor stimulation. PMID- 10386969 TI - Orphanin FQ/nociceptin modulation of mesolimbic dopamine transmission determined by microdialysis. AB - Orphanin FQ has been reported to suppress extracellular dopamine levels in the nucleus accumbens after intracerebroventricular administration. This study sought to provide evidence for an intra-ventral tegmental site of action for this effect using a dual-probe microdialysis experimental design. Orphanin FQ was applied to the ventral tegmental area of anesthetized rats by reverse dialysis while extracellular dopamine was sampled with a second dialysis probe in the nucleus accumbens. Orphanin FQ at a probe concentration of 1 mM (but not at 0.1 mM) significantly reduced nucleus accumbens dialysate dopamine levels. The receptor inactive analogue, des-Phe1-orphanin FQ (1 mM), produced a small but significant increase in nucleus accumbens dialysate dopamine levels. Simultaneous measurement of ventral tegmental area dialysate amino acid content revealed significant increases in both GABA and glutamate during infusion of orphanin FQ (1 mM). To determine if increased GABA overflow mediates the action of orphanin FQ on mesolimbic neurons, orphanin FQ (10 nmol) was microinjected directly into the ventral tegmental area in the presence or absence of the GABA(A) receptor antagonist, bicuculline (1 nmol). Bicuculline transiently blocked the suppressive action of orphanin FQ on accumbens dialysate dopamine levels. These data indicate that orphanin FQ decreases dopamine transmission in the nucleus accumbens by inhibiting dopamine neuronal activity in the ventral tegmental area through a mechanism that may involve an increased overflow of GABA. PMID- 10386970 TI - Increased AP-1 DNA binding activity in PC12 cells treated with lead. AB - The possibility that the mechanism of lead neurotoxicity may be at the level of transcription was investigated in PC12 cells. In electrophoretic mobility gel shift assays Pb2+ was found to increase activator protein-1 complex (AP-1) DNA binding activity in PC12 cells; the increase was time- and concentration dependent. Exposure to Pb2+ also resulted in an increase in AP-1-driven transcription in cerebellar granule cells transfected with a luciferase gene reporter construct. The increase in AP-1 DNA binding activity by Pb2+ required protein synthesis. The increase was mediated by protein kinase C because depletion of protein kinase C and an inhibitor of protein kinase C prevented the increase in AP-1 DNA binding activity by Pb2+. Fra-2 and JunD were found in supershift assays to be the major components of the AP-1 that was increased by Pb2+. In summary, our studies indicate that Pb2+ increases AP-1 DNA binding activity in PC12 cells by a pathway that requires protein kinase C and new protein synthesis. PMID- 10386971 TI - Alzheimer's disease: correlation of the suppression of beta-amyloid peptide secretion from cultured cells with inhibition of the chymotrypsin-like activity of the proteasome. AB - Peptide aldehyde inhibitors of the chymotrypsin-like activity of the proteasome (CLIP) such as N-acetyl-Leu-Leu-Nle-H (or ALLN) have been shown previously to inhibit the secretion of beta-amyloid peptide (A beta) from cells. To evaluate more fully the role of the proteasome in this process, we have tested the effects on A beta formation of a much wider range of peptide-based inhibitors of CLIP than published previously. The inhibitors tested included several peptide boronates, some of which proved to be the most potent peptide-based inhibitors of beta-amyloid production reported so far. We found that the ability of the peptide aldehyde and boronate inhibitors to suppress A beta formation from cells correlated extremely well with their potency as CLIP inhibitors. Thus, we conclude that the proteasome may be involved either directly or indirectly in A beta formation. PMID- 10386972 TI - Temporal patterns of poly(ADP-ribose) polymerase activation in the cortex following experimental brain injury in the rat. AB - The activation of poly(ADP-ribose) polymerase, a DNA base excision repair enzyme, is indicative of DNA damage. This enzyme also undergoes site-specific proteolysis during apoptosis. Because both DNA fragmentation and apoptosis are known to occur following experimental brain injury, we investigated the effect of lateral fluid percussion brain injury on poly(ADP-ribose) polymerase activity and cleavage. Male Sprague-Dawley rats (n = 52) were anesthetized, subjected to fluid percussion brain injury of moderate severity (2.5-2.8 atm), and killed at 30 min, 2 h, 6 h, 24 h, 3 days, or 7 days postinjury. Genomic DNA from injured cortex at 24 h, but not at 30 min, was both fragmented and able to stimulate exogenous poly(ADP-ribose) polymerase. Endogenous poly(ADP-ribose) polymerase activity, however, was enhanced in the injured cortex at 30 min but subsequently returned to baseline levels. Slight fragmentation of poly(ADP-ribose) polymerase was detected in the injured cortex in the first 3 days following injury, but significant cleavage was detected at 7 days postinjury. Taken together, these data suggest that poly(ADP-ribose) polymerase-mediated DNA repair is initiated in the acute posttraumatic period but that subsequent poly(ADP-ribose) polymerase activation does not occur, possibly owing to delayed apoptosis-associated proteolysis, which may impair the repair of damaged DNA. PMID- 10386973 TI - Effect of chronic angiotensin-converting enzyme inhibition on striatal dopamine content in the MPTP-treated mouse. AB - We have previously shown that chronic treatment with the angiotensin-converting enzyme inhibitor perindopril increased striatal dopamine levels by 2.5-fold in normal Sprague-Dawley rats, possibly via modulation of the striatal opioid or tachykinin levels. In the present study, we investigated if this effect of perindopril persists in an animal model of Parkinson's disease, the 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated mouse. C57BL/6 mice were treated with the neurotoxin (30 mg/kg/day intraperitoneally) for 4 days and then left for 3 weeks to allow the degeneration of striatal dopaminergic terminals. At this time, the mice exhibited a 40% decrease in striatal dopamine content and an accompanying 46% increase in dopamine D2 receptor levels compared with control untreated mice. The dopamine content returned to control levels, and the increase in dopamine D2 receptor levels was attenuated in mice treated with perindopril (5 mg/kg/day orally for 7 days) 2 weeks after the last dose of MPTP. When the angiotensin-converting enzyme inhibitor was administered (5 mg/kg/day for 7 days) immediately after the cessation of the MPTP treatment, there was no reversal of the effect of the neurotoxin in decreasing striatal dopamine content. Our results demonstrate that perindopril is an effective agent in increasing striatal dopamine content in an animal model of Parkinson's disease. PMID- 10386974 TI - Depolarization of in situ mitochondria due to hydrogen peroxide-induced oxidative stress in nerve terminals: inhibition of alpha-ketoglutarate dehydrogenase. AB - Mitochondrial membrane potential (delta psi(m)) was determined in intact isolated nerve terminals using the membrane potential-sensitive probe JC-1. Oxidative stress induced by H2O2 (0.1-1 mM) caused only a minor decrease in delta psi(m). When complex I of the respiratory chain was inhibited by rotenone (2 microM), delta psi(m) was unaltered, but on subsequent addition of H2O2, delta psi(m) started to decrease and collapsed during incubation with 0.5 mM H2O2 for 12 min. The ATP level and [ATP]/[ADP] ratio were greatly reduced in the simultaneous presence of rotenone and H2O2. H2O2 also induced a marked reduction in delta psi(m) when added after oligomycin (10 microM), an inhibitor of F0F1-ATPase. H2O2 (0.1 or 0.5 mM) inhibited alpha-ketoglutarate dehydrogenase and decreased the steady-state NAD(P)H level in nerve terminals. It is concluded that there are at least two factors that determine delta psi(m) in the presence of H2O2: (a) The NADH level reduced owing to inhibition of alpha-ketoglutarate dehydrogenase is insufficient to ensure an optimal rate of respiration, which is reflected in a fall of delta psi(m) when the F0F1-ATPase is not functional. (b) The greatly reduced ATP level in the presence of rotenone and H2O2 prevents maintenance of delta psi(m) by F0F1-ATPase. The results indicate that to maintain delta psi(m) in the nerve terminal during H2O2-induced oxidative stress, both complex I and F0F1-ATPase must be functional. Collapse of delta psi(m) could be a critical event in neuronal injury in ischemia or Parkinson's disease when H2O2 is generated in excess and complex I of the respiratory chain is simultaneously impaired. PMID- 10386975 TI - Responses of heat shock proteins hsp27, alphaB-crystallin, and hsp70 in rat brain after kainic acid-induced seizure activity. AB - We determined the changes in the levels of the mammalian small heat shock protein of 25-28 kDa (hsp27) and the hsp alphaB-crystallin in various regions of rat brain after kainic acid-induced seizure activity by means of specific immunoassays. The levels of hsp27 in the hippocampus and entorhinal cortex were markedly increased and reached a maximum (1.5-2 microg/mg of protein) 2-4 days after the seizure. The levels of hsp27 in these regions were considerably high even 10 days after the seizure. A marked increase in levels of mRNA for hsp27 was also observed in the hippocampus of rats 1-2 days after the seizure. A severalfold increase in the levels of alphaB-crystallin was observed in the hippocampus and entorhinal cortex of rats 2 days after the seizure. However, the maximum levels were <50 ng/mg of protein. The levels of protein sulfhydryl group and glutathione were significantly reduced in the hippocampus of rats at 24 h after the seizure, which might have enhanced the expressions of hsp27 and alphaB crystallin. The expression of inducible mammalian hsp of 70 kDa (hsp70) was also enhanced in the hippocampus of rats after the seizure, as detected by western and northern blotting analyses. Immunohistochemically, an intensive staining of hsp27 was observed in both glial cells and neurons in the hippocampus, piriform cortex, and entorhinal cortex of rats with kainic acid-induced seizure. However, in the cerebellum, where the receptors for kainic acid are also rich, hsp27 was barely induced in the same rats. This might be due to high levels of the cerebellar calcium-binding proteins parvalbumin and 28-kDa calbindin-D, which might have a protective effect against the kainic acid-inducible damage. PMID- 10386976 TI - Glutamate neurotoxicity in rat cerebellar granule cells involves cytochrome c release from mitochondria and mitochondrial shuttle impairment. AB - To gain some insight into the mechanism by which glutamate neurotoxicity takes place in cerebellar granule cells, two steps of glucose oxidation were investigated: the electron flow via respiratory chain from certain substrates to oxygen and the transfer of extramitochondrial reducing equivalents via the mitochondrial shuttles. However, cytochrome c release from intact mitochondria was found to occur in glutamate-treated cells as detected photometrically in the supernatant of the cell homogenate suspension. As a result of cytochrome c release, an increase of the oxidation of externally added NADH was found, probably occurring via the NADH-b5 oxidoreductase of the outer mitochondrial membrane. When the two mitochondrial shuttles glycerol 3 phosphate/dihydroxyacetone phosphate and malate/oxaloacetate, devoted to oxidizing externally added NADH, were reconstructed, both were found to be impaired under glutamate neurotoxicity. Consistent early activation in two NADH oxidizing mechanisms, i.e., lactate production and plasma membrane NADH oxidoreductase activity, was found in glutamate-treated cells. In spite of this, the increase in the cell NADH fluorescence was found to be time-dependent, an index of the progressive damage of the cell. PMID- 10386977 TI - Inhibition of peroxynitrite-mediated oxidation of dopamine by flavonoid and phenolic antioxidants and their structural relationships. AB - The interaction between peroxynitrite and dopamine and the inhibition of this reaction by plant-derived antioxidants have been investigated. Peroxynitrite promoted the oxidation of dopamine to 6-hydroxyindole-5-one as characterised by HPLC and photodiode array spectra, akin to the products of the tyrosinase dopamine reaction, but no evidence of dopamine nitration was obtained. Although peroxynitrite did not cause nitration of dopamine in vitro, the catecholamine is capable of inhibiting the formation of 3-nitrotyrosine from peroxynitrite mediated nitration of tyrosine. The plant-derived phenolic compounds, caffeic acid and catechin, inhibited peroxynitrite-mediated oxidation of dopamine. This effect is attributed to the ability of catechol-containing antioxidants to reduce peroxynitrite through electron donation, resulting in their oxidation to the corresponding o-quinones. The antioxidant effect of caffeic acid and catechin was comparable to that of the endogenous antioxidant, glutathione. In contrast, the structurally related monohydroxylated hydroxycinnamates, p-coumaric acid and ferulic acid, which inhibit tyrosine nitration through a mechanism of competitive nitration, did not inhibit peroxynitrite-induced dopamine oxidation. The findings of the present study suggest that certain plant-derived phenolics can inhibit dopamine oxidation. PMID- 10386978 TI - Expression of mouse sialic acid on gangliosides of a human glioma grown as a xenograft in SCID mice. AB - Ganglioside sialic acid content was examined in the U87-MG human glioma grown as cultured cells and as a xenograft in severe combined immunodeficiency (SCID) mice. The cultured cells and the xenograft possessed N-glycolylneuraminic acid (NeuGc)-containing gangliosides, despite the inability of human cells to synthesize NeuGc. Human cells express only N-acetylneuraminic acid (NeuAc) containing gangliosides, whereas mouse cells express both NeuAc- and NeuGc containing gangliosides. Small amounts of NeuGc ganglioside sialic acid (2-3% of total ganglioside sialic acid) were detected in the cultured cells, whereas large amounts (66% of total ganglioside sialic acid) were detected in the xenograft. The NeuGc in gangliosides of the cultured cells was derived from gangliosides in the fetal bovine serum of the culture medium, whereas that in the U87-MG xenograft was derived from gangliosides of the SCID host. The chromatographic distribution of U87-MG gangliosides differed markedly between the in vitro and in vivo growth environments. The neutral glycosphingolipids in the U87-MG cells consisted largely of glucosylceramide, galactosylceramide, and lactosylceramide, and their distribution also differed in the two growth environments. Asialo-GM1 (Gg4Cer) was not present in the cultured tumor cells but was expressed in the xenograft, suggesting an origin from infiltrating cells (macrophages) from the SCID host. The infiltration of mouse host cells and the expression of mouse sialic acid on human tumor cell glycoconjugates may alter the biochemical and immunogenic properties of xenografts. PMID- 10386979 TI - Receptor-mediated transcytosis of cyclophilin B through the blood-brain barrier. AB - Cyclophilin B (CyPB) is a cyclosporin A (CsA)-binding protein mainly located in intracellular vesicles and secreted in biological fluids. In previous works, we demonstrated that CyPB interacts with T lymphocytes and enhances in vitro cellular incorporation and activity of CsA. In addition to its immunosuppressive activity, CsA is able to promote regeneration of damaged peripheral nerves. However, the crossing of the drug from plasma to neural tissue is restricted by the relative impermeability of the blood-brain barrier. To know whether CyPB might also participate in the delivery of CsA into the brain, we have analyzed the interactions of CyPB with brain capillary endothelial cells. First, we demonstrated that CyPB binds to two types of binding sites present at the surface of capillary endothelial cells from various species of tissues. The first type of binding sites (K(D) = 300 nM; number of sites = 3 x 10(6)) is related to interactions with negatively charged compounds such as proteoglycans. The second type of binding sites, approximately 50,000 per cell, exhibits a higher affinity for CyPB (K(D) = 15 nM) and is involved in an endocytosis process, indicating it might correspond to a functional receptor. Finally, the use of an in vitro model of blood-brain barrier allowed us to demonstrate that CyPB is transcytosed by a receptor-mediated pathway (flux = 16.5 fmol/cm2/h). In these conditions, CyPB did not significantly modify the passage of CsA, indicating that it is unlikely to provide a pathway for CsA brain delivery. PMID- 10386980 TI - Alterations in ionized and total blood magnesium after experimental traumatic brain injury: relationship to neurobehavioral outcome and neuroprotective efficacy of magnesium chloride. AB - Experimental evidence suggests that magnesium plays a role in the pathophysiological sequelae of brain injury. The present study examined the variation of blood ionized and total magnesium, as well as potassium, sodium, and ionized calcium, after experimental fluid percussion brain injury in rats. Blood ionized magnesium concentration significantly declined from 0.45 +/- 0.02 to 0.32 +/- 0.02 mM by 30 min postinjury and stayed depressed for the 24-h study period in vehicle-treated rats. Blood total magnesium concentration was 0.59 +/- 0.01 mM and remained stable over time in brain-injured vehicle-treated animals. When magnesium chloride (125 micromol/rat) was administered 1 h postinjury, ionized magnesium levels were restored by 2 h postinjury and remained at normal values up to 24 h following brain trauma. Magnesium treatment also significantly reduced posttraumatic neuromotor impairments 1 and 2 weeks after the insult, but failed to attenuate spatial learning deficits. A significant positive and linear correlation could be established between ionized magnesium levels measured 24 h postinjury and neuromotor outcome at 1 and 2 weeks. We conclude that acute ionized magnesium measurement may be a predictor of long-term neurobehavioral outcome following head injury and that delayed administration of magnesium chloride can restore blood magnesium concentration and attenuate neurological motor deficits in brain-injured rats. PMID- 10386981 TI - The cholecystokininB receptor is coupled to two effector pathways through pertussis toxin-sensitive and -insensitive G proteins. AB - Previous binding studies have suggested the existence of two affinity states for type B cholecystokinin receptors (CCK(B)R), which could correspond to different coupling states of the receptor to G proteins. To test this hypothesis, we have further investigated signal transduction pathways coupled to rat CCK(B)R stably transfected in Chinese hamster ovary cells. We show that CCK(B)R are coupled to two distinct transduction pathways involving two different G proteins, a pertussis toxin-insensitive/phospholipase C pathway leading to the production of inositol phosphate and arachidonic acid, and a pertussis toxin sensitive/phospholipase A2 pathway leading to the release of arachidonic acid. We further demonstrate that the relative degree of activation of each effector pathway by different specific CCK(B)R agonists is the same, and that a specific CCK(B)R antagonist, RB213, can differentially antagonize the two signal transduction pathways elicited by these agonists. Taken all together, these data could be explained by the recently proposed theory assuming that the receptor can exist in a three-state model in which two active conformations corresponding to the complex formed by the receptor with two different G proteins coexist. According to this model, agonists or antagonists could recognize preferentially either conformation of the activated receptor, leading to variable behavior in a system containing a single receptor type. PMID- 10386982 TI - Membrane microviscosity modulates mu-opioid receptor conformational transitions and agonist efficacy. AB - The influence of membrane microviscosity on mu-opioid agonist and antagonist binding, as well as agonist efficacy, was examined in membranes prepared from SH SY5Y cells and from a C6 glioma cell line stably expressing the rat mu-opioid receptor (C6mu). Addition of cholesteryl hemisuccinate (CHS) to cell membranes increased membrane microviscosity and reduced the inhibitory effect of sodium and guanine nucleotides on the affinity of the full agonists sufentanil and [D-Ala2,N MePhe4,Gly-ol5]enkephalin (DAMGO) for the mu-opioid receptor. Binding of the antagonists [3H]naltrexone and [3H]diprenorphine and the partial agonist nalbuphine was unaffected by CHS. The effect of CHS on agonist binding was reversed by subsequent addition of cis-vaccenic acid, suggesting that the effect of CHS is the result of increased membrane microviscosity and not a specific sterol-receptor interaction. CHS addition increased the potency of DAMGO to stimulate guanosine-5'-O-(3-[35S]thio)triphosphate binding by fourfold, whereas the potency of nalbuphine was unaffected. However, nalbuphine efficacy relative to that of the full agonist DAMGO was strongly increased in CHS-treated membranes compared with that in control membranes. Membrane rigidification also resulted in an increased efficacy for the partial agonists meperidine, profadol, and butorphanol relative to that of DAMGO as measured by agonist-stimulated GTPase activity in control and CHS-modified membranes. These findings support a regulatory role for membrane microviscosity in receptor-mediated G protein activation. PMID- 10386983 TI - A globin fragment, LVV-hemorphin-7, induces [3H]thymidine incorporation in a neuronal cell line via the AT4 receptor. AB - The AT4 receptor was characterized initially as a specific binding site for angiotensin IV, a C-terminal fragment of the vasoactive peptide angiotensin II. Recently, we found that LVV-hemorphin-7, a fragment of beta globin, is an abundant peptide in the brain and binds to the AT4 receptor with high affinity and specificity. In the neuroblastoma/glioma hybrid cell line, NG108-15, LVV hemorphin-7 and angiotensin IV competed for 125I-angiotensin IV binding in a biphasic fashion with IC50 values of 1.2 x 10(-10) and 1.1 x 10(-9) M for the high-affinity site, respectively, and 6.7 x 10(-8) and 1.5 x 10(-8) M for the low affinity site, respectively. Both peptides were internalized rapidly by the cells. However, LVV-hemorphin-7, but not angiotensin IV, elicited a 1.8-fold increase in DNA synthesis in a dose-dependent manner. Furthermore, co-incubation of the cells with an excess of angiotensin IV (10(-6) M) inhibited LVV-hemorphin 7-stimulated DNA synthesis. Therefore, whereas LVV-hemorphin-7 and angiotensin IV were capable of binding to the AT4 receptor, only LVV-hemorphin-7 elicited [3H]thymidine incorporation in NG108-15 cells. In contrast, angiotensin IV behaved as an antagonist. The current finding suggests that LVV-hemorphin-7 is a functional peptide in the central nervous system and in view of its abundance in neural tissue, compared with angiotensin IV, may be of significant physiological importance. PMID- 10386984 TI - Effects of systemic immunogenic insults and circulating proinflammatory cytokines on the transcription of the inhibitory factor kappaB alpha within specific cellular populations of the rat brain. AB - Expression of the inhibitory factor kappaB alpha (IkappaB alpha) reflects the activity of nuclear factor kappaB(NF-kappaB) and is a powerful tool to investigate the regulation of the transcription factor within the CNS. IkappaB alpha mRNA was evaluated in the rat brain by means of in situ hybridization following different immunogenic stimuli; i.e., intraperitoneal (i.p.) and intravenous (i.v.) lipopolysaccharide (LPS), i.v. recombinant rat interleukin (IL) 1beta, IL-6, or tumor necrosis factor-alpha (TNF-alpha), and intramuscular (i.m.) turpentine injection, used here as a model of systemic localized inflammatory insult. Systemic LPS, IL-1beta, and TNF-alpha caused a rapid and transient transcriptional activation of IkappaB alpha along the blood vessels of the entire brain; the signal was very intense 30-60 min after the i.v. injections and returned to undetectable levels from 2 to 12 h depending on the challenge. Double-labeling procedure provided the anatomical evidence that IkappaB alpha expressing cells within the microvasculature were essentially of the endothelial type, as they were immunoreactive to the von Willebrand factor. Scattered small cells were also found across the brain of LPS-, IL-1beta-, and TNF-alpha-injected rats at time 1-3 h, and microglial (OX-42)-immunoreactive cells were positive for the transcript. Such expression within parenchymal microglia was nevertheless not observed in the brain following a localized and sterile inflammatory insult. Indeed, i.m. turpentine administration stimulated IkappaB alpha transcription quite uniquely within the endothelium of the brain capillaries, an effect that paralleled the swelling of the injection site and lasted up to 24 h after the aggression. In contrast to these immunogenic challenges, i.v. IL-6 injection failed to activate the gene encoding IkappaB alpha in the rat brain. These results indicate that NF-kappaB may play a crucial role in specific cellular populations of the CNS to trigger transcription of immune-related genes and that IkappaB alpha resynthesis may act as a dynamic intracellular inhibitory feedback to avoid exaggeration of the response. It is possible that IkappaB alpha expression in cells of the blood-brain barrier is a general mechanism that takes place during systemic inflammation, whereas the participation of NF-kappaB related molecules within parenchymal cells of the CNS is solicited during more severe conditions such as blood sepsis and endotoxemia. PMID- 10386985 TI - Activation of caspase-3 in developmental models of programmed cell death in neurons of the substantia nigra. AB - Programmed cell death has been proposed to play a role in the death of neurons in acute and chronic degenerative neurologic disease. There is now evidence that the caspases, a family of cysteine proteases, mediate programmed cell death in various cells. In neurons, caspase-3 (CPP32/Yama/apopain), in particular, has been proposed to play a role. We examined the expression of caspase-3 in three models of programmed cell death affecting neurons of the substantia nigra in the rat: natural developmental neuron death and induced developmental death following either striatal target injury with quinolinic acid or dopamine terminal lesion with intrastriatal injection of 6-hydroxydopamine. Using an antibody to the large (p17) subunit of activated caspase-3, we have found that activated enzyme is expressed in apoptotic profiles in all models. Increased p17 immunostaining correlated with increased enzyme activity. The subcellular distribution of activated caspase-3 differed among the models: In natural cell death and the target injury model, it was strictly nuclear, whereas in the toxin model, it was also cytoplasmic. We conclude that p17 immunostaining is a useful marker for programmed cell death in neurons of the substantia nigra. PMID- 10386986 TI - ATP-stimulated Ca2+ influx and phospholipase D activities of a rat brain-derived type-2 astrocyte cell line, RBA-2, are mediated through P2X7 receptors. AB - This study characterizes and examines the P2 receptor-mediated signal transduction pathway of a rat brain-derived type 2 astrocyte cell line, RBA-2. ATP induced Ca2+ influx and activated phospholipase D (PLD). The ATP-stimulated Ca2+ influx was inhibited by pretreating cells with P2 receptor antagonist, pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS), in a concentration dependent manner. The agonist 2'- and 3'-O-(4-benzoylbenzoyl)adenosine 5' triphosphate (BzATP) stimulated the largest increases in intracellular Ca2+ concentrations ([Ca2+]i); ATP, 2-methylthioadenosine triphosphate tetrasodium, and ATPgammaS were much less effective, whereas UTP, ADP, alpha,beta-methylene ATP, and beta,gamma-methylene-ATP were ineffective. Furthermore, removal of extracellular Mg2+ enhanced the ATP- and BzATP-stimulated increases in [Ca2+]i. BzATP stimulated PLD in a concentration- and time-dependent manner that could be abolished by removal of extracellular Ca2+ and was inhibited by suramin, PPADS, and oxidized ATP. In addition, PLD activities were activated by the Ca2+ mobilization agent, ionomycin, in an extracellular Ca2+ concentration-dependent manner. Both staurosporine and prolonged phorbol ester treatment inhibited BzATP stimulated PLD activity. Taken together, these data indicate that activation of the P2X7 receptors induces Ca2+ influx and stimulates a Ca2+-dependent PLD in RBA 2 astrocytes. Furthermore, protein kinase C regulates this PLD. PMID- 10386988 TI - Activation of nuclear factor kappaB by nitric oxide in rat striatal neurones: differential inhibition of the p50 and p65 subunits by dexamethasone. AB - Nitric oxide (NO), an intercellular messenger in the brain, has been implicated in both neuronal plasticity and neurotoxicity. It has been suggested that NO can activate the DNA binding activity of nuclear factor kappaB (NF-kappaB) family proteins in some cell types while having an inhibitory effect in others. In this study we have investigated the effect of acute NO in primary neuronal cultures of rat striatum using immunohistochemistry. Exposure of neurones to the NO-mimetic S nitroso-n-acetylpenicillamine (SNAP; 200 microM) and to bacterial lipopolysaccharide (LPS; 10 microg/ml) for 30 min increased nuclear protein expression of the p50 subunit of NF-kappaB. SNAP also enhanced nuclear protein expression of the p65 subunit of NF-kappaB. Simultaneously, the cytoplasmic expression of phosphorylated inhibitory protein IkappaB alpha was dramatically increased by SNAP (200 microM), LPS (10 microg/ml), and kainate (50 microM) treatment. In the adult rat, stimulation with NOR-3 (2 mg/kg), a NO donor, increased NF-kappaB DNA binding activity in the striatum after 45 min. Because glucocorticoids inhibit NF-kappaB activity, primary cultures were pretreated with dexamethasone (50 microM) before SNAP, LPS, and kainate treatment, and the effect on the protein expression level of the individual subunits p50 and p65 present in the classical form of the transcription factor NF-kappaB was assessed. Dexamethasone pretreatment resulted in a marked reduction of p65 protein in striatal neurones after SNAP, LPS, and kainate, whereas p50 expression was reduced by dexamethasone pretreatment only after an LPS stimulus. This study indicates that NO-releasing compounds can directly induce nuclear NF-kappaB subunit expression in rat striatum and that glucocorticoids selectively inhibit p65 subunit expression following exposure to NO. PMID- 10386987 TI - Systemic administration of NMDA and AMPA receptor antagonists reverses the neurochemical changes induced by nigrostriatal denervation in basal ganglia. AB - In Parkinson's disease, nigrostriatal denervation leads to an overactivity of the subthalamic nucleus and its target areas, which is responsible of the clinical manifestations of the disease. Because the subthalamic nucleus uses glutamate as neurotransmitter and is innervated by glutamatergic fibers, pharmacological blockade of glutamate transmission might be expected to restore the cascade of neurochemical changes induced by a dopaminergic denervation within the basal ganglia. To test this hypothesis, two types of glutamate antagonists, the NMDA receptor antagonist MK-801 and the alpha-amino-3-hydroxy-5-methylisoxazole-4 propionate (AMPA) receptor antagonist LY293558, were administered systemically, either alone or in combination with L-DOPA, in rats with a unilateral 6 hydroxydopamine lesion of the nigrostriatal dopamine pathway. The effect of treatment was assessed neurochemically by analyzing at the cellular level the functional activity of basal ganglia output structures and the subthalamic nucleus using the expression levels of the mRNAs coding for glutamic acid decarboxylase and cytochrome oxidase, respectively, as molecular markers of neuronal activity. The present study shows that treatment with glutamate antagonists, and particularly with AMPA antagonists, alone or in combination with L-DOPA, reverses the overactivity of the subthalamic nucleus and its target areas induced by nigrostriatal denervation. These results furnish the neurochemical basis for the potential use of glutamate antagonists as therapeutic agents in Parkinson's disease. PMID- 10386989 TI - Biochemical and neurotoxicological effects of L-2-chloropropionic acid on rodent brain. AB - L-2-Chloropropionic acid (L-CPA) is selectively toxic to cerebellar granule cells; necrosis is first observed in rats 36 h after L-CPA administration (750 mg/kg p.o.) and becomes marked by 48 h. L-CPA has also been shown to activate the mitochondrial pyruvate dehydrogenase (PDH) complex in fasted adult rats, resulting in reduced blood glucose and lactate levels. This study aimed to investigate the biochemical and neurotoxicological effects of L-CPA on the brain. Extracts, prepared from guinea-pig cerebellar and cerebral cortex slices incubated in the presence of L-CPA, were analysed using 1H magnetic resonance spectroscopy, 31P magnetic resonance spectroscopy, and amino acid analysis. Glucose metabolism was studied by monitoring the metabolism of [1-(13)C]glucose using gas chromatography/mass spectrometry. Increased glucose metabolism and decreases in the pool sizes of lactate and alanine were observed in both tissues, demonstrating activation of the PDH complex. Extracts were also prepared from the forebrain and cerebellum of animals that had been treated in vivo with L-CPA and analysed as described for the in vitro studies. Similar evidence for PDH activation was demonstrated at 2 and 24 h after dosing in both tissues. At 48 h after dosing, when signs of toxicity are observed, an increase in the lactate concentration and a decrease in N-acetylaspartate in the cerebellum but not in the forebrain confirmed the selective neurotoxic action of L-CPA. These results suggest that activation of the PDH complex does not directly lead to the delayed selective neurotoxicity of L-CPA. PMID- 10386990 TI - Internalization and proteolytic action of botulinum toxins in CNS neurons and astrocytes. AB - Tetanus and botulinum toxins bind and are internalized at the neuromuscular junction. Botulinum neurotoxins (BoNTs) enter the cytosol at the motor nerve terminal; tetanus neurotoxin (TeNT) proceeds retroaxonally inside the motor axon to reach the spinal cord inhibitory interneurons. Although the major target of BoNTs is the peripheral cholinergic terminals, CNS neurons are susceptible to intoxication as well. We investigated the route of entry and the proteolytic activity of BoNT/B and BoNT/F in cultured hippocampal neurons and astrocytes. We show that, differently from TeNT, which enters hippocampal neurons via the process of synaptic vesicle (SV) recycling, BoNTs are internalized and cleave the substrate synaptobrevin/VAMP2 via a process independent of synaptic activity. Labeling of living neurons with Texas Red-conjugated BoNTs and fluoresceinated dextran revealed that these toxins enter hippocampal neurons via endocytic processes not mediated by SV recycling. Botulinum toxins also exploit endocytosis to enter cultured astrocytes, where they partially cleave cellubrevin, a ubiquitous synaptobrevin/VAMP isoform. These results indicate that, in spite of their closely related protein structure, TeNT and BoNTs use different routes to penetrate hippocampal neurons. These findings bear important implications for the identification of the protein receptors of clostridial toxins. PMID- 10386991 TI - Molecular characterization and imidacloprid selectivity of nicotinic acetylcholine receptor subunits from the peach-potato aphid Myzus persicae. AB - The recent introduction of the chloronicotinyl insecticide imidacloprid, targeting insect nicotinic acetylcholine receptors (nAChRs), emphasises the importance of a detailed molecular characterisation of these receptors. We are investigating the molecular diversity of insect nAChR subunit genes in an important agricultural pest, the peach-potato aphid Myzus persicae. Two M. persicae alpha-subunit cDNAs, Mp alpha1 and Mp alpha2, have been cloned previously. Here we report the isolation of three novel alpha-subunit genes (Mp alpha3-5) with overall amino acid sequence identities between 43 and 76% to characterised insect nAChR subunits. Alignment of their amino acid sequences with other invertebrate and vertebrate nAChR subunits suggests that the insect alpha subunits evolved in parallel to the vertebrate neuronal nAChRs and that the insect non-alpha subunits are clearly different from vertebrate neuronal beta and muscle non-alpha subunits. The discovery of novel subtypes in M. persicae is a further indicator of the complexity of the insect nAChR gene family. Heterologous co-expression of M. persicae nAChR alpha-subunit cDNAs with the rat beta2 in Drosophila S2 cells resulted in high-affinity binding of nicotinic radioligands. The affinity of recombinant nAChRs for [3H]imidacloprid was influenced strongly by the alpha subtype. This is the first demonstration that imidacloprid selectively acts on Mp alpha2 and Mp alpha3 subunits, but not Mp alpha1, in M. persicae. PMID- 10386992 TI - GSH transport in immortalized mouse brain endothelial cells: evidence for apical localization of a sodium-dependent GSH transporter. AB - We have previously shown GSH transport across the blood-brain barrier in vivo and expression of transport in Xenopus laevis oocytes injected with bovine brain capillary mRNA. In the present study, we have used MBEC-4, an immortalized mouse brain endothelial cell line, to establish the presence of Na+-dependent and Na+ independent GSH transport and have localized the Na+-dependent transporter using domain-enriched plasma membrane vesicles. In cells depleted of GSH with buthionine sulfoximine, a significant increase of intracellular GSH could be demonstrated only in the presence of Na+. Partial but significant Na+ dependency of [35S]GSH uptake was observed for two GSH concentrations in MBEC-4 cells in which gamma-glutamyltranspeptidase and gamma-glutamylcysteine synthetase were inhibited to ensure absence of breakdown and resynthesis of GSH. Uniqueness of Na+-dependent uptake in MBEC-4 cells was confirmed with parallel uptake studies with Cos-7 cells that did not show this activity. Molecular form of uptake was verified as predominantly GSH, and very little conversion of [35S]cysteine to GSH occurred under the same incubation conditions. Poly(A)+ RNA from MBEC expressed GSH uptake with significant (approximately 40-70%) Na+ dependency, whereas uptake expressed by poly(A)+ RNA from HepG2 and Cos-1 cells was Na+ independent. Plasma membrane vesicles from MBEC were separated into three fractions (30, 34, and 38% sucrose, by wt) by density gradient centrifugation. Na+-dependent glucose transport, reported to be localized to the abluminal membrane, was found to be associated with the 38% fraction (abluminal). Na+-dependent GSH transport was present in the 30% fraction, which was identified as the apical (luminal) membrane by localization of P-glycoprotein 170 by western blot analysis. Localization of Na+-dependent GSH transport to the luminal membrane and its ability to drive up intracellular GSH may find application in the delivery of supplemented GSH to the brain in vivo. PMID- 10386994 TI - A novel two-site enzyme immunoassay reveals the regional distributions of and developmental changes in GluR1 and NMDAR1 protein contents in the rat brain. AB - Glutamate receptors, including the alpha-amino-3-hydroxy-4-methylisoxazole-4 propionic acid (AMPA) and NMDA receptors, play an important role in neural development and synaptic plasticity in the brain. To date, it has been difficult to correlate accurately individual biochemical phenomena with quantitative and qualitative changes in receptors occurring in specific neurons or synapses. In the present study, we established a two-site enzyme immunoassay for two key subunits of the AMPA and NMDA receptors. Its sensitivities were extremely high, 30 pg for GluR1 and 15 pg for the NMDAR1 receptor containing the C2 exon [NMDAR1(C2)], which enabled us to measure their contents in a few milligrams of hippocampal tissue. Regional and developmental variations in receptor protein levels were much more marked than those reported for mRNA: The absolute GluR1 protein content was highest in the rat hippocampus, whereas the NMDAR1(C2) content was high in all the forebrain regions examined. GluR1 protein levels increased most markedly during the second and third weeks of postnatal life, whereas NMDAR1(C2) content increased during the first postnatal week. In the adult rat brain, the ratio of GluR1 protein to NMDAR1 protein was markedly lower in neocortical regions (approximately 2%) and the highest in cerebellum (22%). Therefore, this two-site enzyme immunoassay is a specific and unique method that enables us to measure absolute tissue contents of the glutamate receptors and will lead to further important discoveries on the biochemical alterations of these receptors. PMID- 10386993 TI - L-Glutamate and insulin enhance glycogen synthesis in cultured astrocytes from the rat brain through different intracellular mechanisms. AB - The effects of L-glutamate and insulin on glycogen synthesis in astrocytes were examined. L-Glutamate and insulin both stimulated glycogen synthesis in primary cultures of rat astrocytes in a dose-dependent manner, as measured by the incorporation of 14C from [14C]glucose into glycogen. D-Aspartate also increased the incorporation of 14C into glycogen. When insulin and L-glutamate were added together, the glycogen synthesis as well as glycogen content of the cells was additively increased. Wortmannin, an inhibitor of phosphatidylinositol 3-kinase, had little effect on glycogen synthesis induced by L-glutamate, whereas it suppressed the insulin-induced glycogen synthesis. These results suggest that the insulin- and L-glutamate-induced glycogen syntheses are mediated by different intracellular mechanisms. In fact, insulin stimulated the conversion of glycogen synthase b to glycogen synthase a, which was suppressed by wortmannin. L Glutamate and D-aspartate, however, did not increase the level of glycogen synthase a activity. By contrast, L-glutamate increased 2-deoxy-D-[3H]glucose uptake by the astrocytes, whereas insulin did not affect the uptake. These results suggest that insulin stimulates glycogen synthesis in astrocytes by activating glycogen synthase, which is dependent on a wortmannin-sensitive signaling pathway. L-Glutamate, however, enhances the glucose uptake, which contributes to the increase in glycogen synthesis in the cells. PMID- 10386995 TI - Glycosylation sites flank phosphorylation sites on synapsin I: O-linked N acetylglucosamine residues are localized within domains mediating synapsin I interactions. AB - Synapsin I is concentrated in nerve terminals, where it appears to anchor synaptic vesicles to the cytoskeleton and thereby ensures a steady supply of fusion-competent synaptic vesicles. Although phosphorylation-dependent binding of synapsin I to cytoskeletal elements and synaptic vesicles is well characterized, little is known about synapsin I's O-linked N-acetylglucosamine (O-GlcNAc) modifications. Here, we identified seven in vivo O-GlcNAcylation sites on synapsin I by analysis of HPLC-purified digests of rat brain synapsin I. The seven O-GlcNAcylation sites (Ser55, Thr56, Thr87, Ser516, Thr524, Thr562, and Ser576) in synapsin I are clustered around its five phosphorylation sites in domains B and D. The proximity of phosphorylation sites to O-GlcNAcylation sites in the regulatory domains of synapsin I suggests that O-GlcNAcylation may modulate phosphorylation and indirectly affect synapsin I interactions. With use of synthetic peptides, however, the presence of an O-GlcNAc at sites Thr562 and Ser576 resulted in only a 66% increase in the Km of calcium/calmodulin-dependent protein kinase II phosphorylation of site Ser566 with no effect on its Vmax. We conclude that O-GlcNAcylation likely plays a more direct role in synapsin I interactions than simply modulating the protein's phosphorylation. PMID- 10386996 TI - Expression of the SM-20 gene promotes death in nerve growth factor-dependent sympathetic neurons. AB - Sympathetic neurons undergo apoptosis when deprived of nerve growth factor (NGF). Inhibitors of RNA or protein synthesis block this death, suggesting that gene expression is important for apoptosis in this system. We have identified SM-20 as a new gene that increases in expression in sympathetic neurons after NGF withdrawal. Expression of SM-20 also increases during neuronal death caused by cytosine arabinoside or the phosphatidylinositol 3-kinase inhibitor LY294002. In addition, SM-20 protein synthesis is elevated in NGF-deprived neurons compared with neurons maintained with NGF. Importantly, expression of SM-20 in sympathetic neurons causes cell death in the presence of NGF. These results suggest that SM 20 may function to regulate cell death in neurons. PMID- 10386997 TI - Sp1/Egr1 motif: a new candidate in the regulation of rat tyrosine hydroxylase gene transcription by immobilization stress. AB - The rat tyrosine hydroxylase (TH) promoter contains a GC box (Sp1 motif) whose function is currently unclear. In this study, we examined the effect of immobilization (IMO) stress on binding of adrenomedullary transcription factors to that site. DNase I footprinting analysis reveals the binding of proteins to the Sp1 motif. Extracts from the adrenal medulla of IMO-treated rats generate a footprint on the Sp1 motif that is smaller than that generated by control extracts. Electrophoretic mobility shift assays using an oligonucleotide containing the Sp1 region show that two Sp1 protein-containing complexes (I and III) form with control extracts. In contrast, extracts from the adrenal medulla of IMO-treated rats form a novel complex (II) that contains the Egr1 protein. This is the first report to reveal changes in the binding pattern on the TH Sp1 motif in response to stress and to identify an overlapping Egr1 site. PMID- 10386998 TI - Antibodies to CSF tau. PMID- 10387000 TI - Effect of protein hydration on receptor conformation: decreased levels of bound water promote metarhodopsin II formation. AB - Neutral solutes were used to investigate the effects of osmotic stress both on the ability of rhodopsin to undergo its activating conformation change and on acyl chain packing in the rod outer segment (ROS) disk membrane. The equilibrium concentration of metarhodopsin II (MII), the conformation of photoactivated rhodopsin, which binds and activates transducin, was increased by glycerol, sucrose, and stachyose in a manner which was linear with osmolality. Analysis of this shift in equilibrium in terms of the dependence of ln(Keq) on osmolality revealed that 20 +/- 1 water molecules are released during the MI-to-MII transition at 20 degrees C, and at 35 degrees C 13 +/- 1 waters are released. At 35 degrees C the average time constant for MII formation was increased from 1.20 +/- 0.09 ms to 1.63 +/- 0.09 ms by addition of 1 osmolal sucrose or glycerol. The effect of the neutral solutes on acyl chain packing in the ROS disk membrane was assessed via measurements of the fluorescence lifetime and anisotropy decay of 1,6-diphenyl-1,3,5-hexatriene (DPH). Analysis of the anisotropy decay of DPH in terms of the rotational diffusion model showed that the angular width of the equilibrium orientational distribution of DPH about the membrane normal was progressively narrowed by increased osmolality. The parameter fv, which is proportional to the overlap between the DPH orientational probability distribution and a random orientational distribution, was reduced by the osmolytes in a manner which was linear with osmolality. This study highlights the potentially opposing interplay between the effect of membrane surface hydration on both the lipid bilayer and integral membrane protein structure. Our results further demonstrate that the binding and release of water molecules play an important role in modulating functional conformational changes for integral membrane proteins, as well as for soluble globular proteins. PMID- 10386999 TI - The A beta peptide of Alzheimer's disease directly produces hydrogen peroxide through metal ion reduction. AB - Oxidative stress markers characterize the neuropathology both of Alzheimer's disease and of amyloid-bearing transgenic mice. The neurotoxicity of amyloid A beta peptides has been linked to peroxide generation in cell cultures by an unknown mechanism. We now show that human A beta directly produces hydrogen peroxide (H2O2) by a mechanism that involves the reduction of metal ions, Fe(III) or Cu(II), setting up conditions for Fenton-type chemistry. Spectrophotometric experiments establish that the A beta peptide reduces Fe(III) and Cu(II) to Fe(II) and Cu(I), respectively. Spectrochemical techniques are used to show that molecular oxygen is then trapped by A beta and reduced to H2O2 in a reaction that is driven by substoichiometric amounts of Fe(II) or Cu(I). In the presence of Cu(II) or Fe(III), A beta produces a positive thiobarbituric-reactive substance (TBARS) assay, compatible with the generation of the hydroxyl radical (OH.). The amounts of both reduced metal and TBARS reactivity are greatest when generated by A beta 1-42 >> A beta 1-40 > rat A beta 1-40, a chemical relationship that correlates with the participation of the native peptides in amyloid pathology. These findings indicate that the accumulation of A beta could be a direct source of oxidative stress in Alzheimer's disease. PMID- 10387001 TI - Nonenzymatic reduction of nitro derivative of a heterocyclic amine IQ by NADH and Cu(II) leads to oxidative DNA damage. AB - Nitro derivative (nitro-IQ) of a carcinogenic heterocyclic amine 2-amino-3 methylimidazo[4,5-f]quinoline (IQ) is known to be a potent mutagen as well as IQ, and nitro-IQ is believed to be activated enzymatically by nitroreductase. We investigated nonenzymatic reduction of nitro-IQ by an endogenous reductant NADH and the ability of inducing DNA damage by nitro-IQ. Nitro-IQ caused DNA damage including 8-oxo-7,8-dihydro-2'-deoxyguanosine in the presence of NADH and Cu(II). Catalase and bathocuproine, a Cu(I)-specific chelator, inhibited the DNA damage, suggesting the involvement of H2O2 and Cu(I). Nitro-IQ induced DNA cleavage frequently at thymine and cytosine residues in the presence of NADH and Cu(II). UV-vis spectroscopic study showed that no spectral change of Nitro-IQ and NADH was observed in the absence of Cu(II), while rapid spectral change was observed in the presence of Cu(II), suggesting that Cu(II) mediated redox reaction of nitro-IQ and NADH. These results suggest that nitro-IQ can be reduced nonenzymatically by NADH in the presence of Cu(II), and the redox reaction resulted in oxidative DNA damage due to the copper-oxygen complex, derived from the reaction of Cu(I) with H2O2. We conclude that nonenzymatic reduction of nitro IQ and resulting in oxidative DNA damage can play a role in carcinogenesis of IQ. PMID- 10387002 TI - Structure of tropinone reductase-II complexed with NADP+ and pseudotropine at 1.9 A resolution: implication for stereospecific substrate binding and catalysis. AB - Tropinone reductase-II (TR-II) catalyzes the NADPH-dependent reduction of the carbonyl group of tropinone to a beta-hydroxyl group. The crystal structure of TR II complexed with NADP+ and pseudotropine (psi-tropine) has been determined at 1.9 A resolution. A seven-residue peptide near the active site, disordered in the unliganded structure, is fixed in the ternary complex by participation of the cofactor and substrate binding. The psi-tropine molecule is bound in an orientation which satisfies the product configuration and the stereochemical arrangement toward the cofactor. The substrate binding site displays a complementarity to the bound substrate (psi-tropine) in its correct orientation. In addition, electrostatic interactions between the substrate and Glu156 seem to specify the binding position and orientation of the substrate. A comparison between the active sites in TR-II and TR-I shows that they provide different van der Waals surfaces and electrostatic features. These differences likely contribute to the correct binding mode of the substrates, which are in opposite orientations in TR-II and TR-I, and to different reaction stereospecificities. The active site structure in the TR-II ternary complex also suggests that the arrangement of the substrate, cofactor, and catalytic residues is stereoelectronically favorable for the reaction. PMID- 10387003 TI - Interchange of catalytic activity within the 2-enoyl-coenzyme A hydratase/isomerase superfamily based on a common active site template. AB - The structures and chemical pathways associated with the members of the 2-enoyl CoA hydratase/isomerase enzyme superfamily are compared to show that a common active site design provides the members of this family with a CoA binding site, an expandable acyl binding pocket, an oxyanion hole for binding/polarizing the thioester C=O, and multiple active site stations for the positioning of acidic and basic amino acid side chains for use in proton shuttling. It is hypothesized that this active site template can be tailored to catalyze a wide range of chemical transformations through strategic positioning of acid/base residues among the active site stations. To test this hypothesis, the active site of one member of the 2-enoyl-CoA hydratase/isomerase family, 4-chlorobenzoyl-CoA dehalogenase, was altered by site-directed mutagenesis to include the two glutamate residues functioning in acid/base catalysis in a second family member, crotonase. Catalysis of the syn hydration of crotonyl-CoA, absent in the wild type 4-chlorobenzoyl-CoA dehalogenase, was shown to occur with the structurally modified 4-chlorobenzoyl-CoA dehalogenase at kcat = 0.06 s-1 and Km = 50 microM. PMID- 10387004 TI - Disulfide bridges in interleukin-8 probed using non-natural disulfide analogues: dissociation of roles in structure from function. AB - The structural and functional roles of the two disulfide bridges in interleukin-8 (IL-8) were addressed using IL-8 analogues with covalently modified disulfide bridges. The analogues were prepared using chemical synthesis by replacement of a cysteine for either homocysteine, penicillamine, or selenocysteine and on folding resulted in a covalently modified disulfide. Deletion of either of the two disulfide bridges by replacement of either cysteine pair with alanine resulted in loss of both structure and function. In contrast, all of the analogues with modified disulfide bridges had native tertiary fold as determined by nuclear magnetic resonance spectroscopic methods. Their structural similarity provided a rational basis for assessing the functional effects of the changes to the disulfide. Modification to the disulfide bridge between cysteines 9 and 50 had only a modest effect on IL-8 function. In contrast, alterations to the 7-34 disulfide bridge resulted in a dramatic reduction in biological potency. Thus, although both disulfide bridges are required for maintenance of the native tertiary fold, their role in determining IL-8 activity is distinct. We propose that 7-34 disulfide has a direct role in determining receptor binding and activation, whereas the 9-50 was not directly involved. The synthesis of non natural disulfide analogues is a novel general approach to structure-activity relationships of disulfide bridges. The demonstration that the participation of disulfide bridges in function can be dissociated from their effects on the stability of the tertiary structure suggests that this method will lead to increased understanding of the roles of disulfide bridges in proteins. PMID- 10387005 TI - Evidence that pcpA encodes 2,6-dichlorohydroquinone dioxygenase, the ring cleavage enzyme required for pentachlorophenol degradation in Sphingomonas chlorophenolica strain ATCC 39723. AB - An enzyme that catalyzes an Fe2+-dependent reaction of 2, 6-dichlorohydroquinone with O2 has been isolated from Sphingomonas chlorophenolica sp. strain ATCC 39723, a soil microorganism capable of complete mineralization of pentachlorophenol. The product of the reaction is too unstable to allow spectroscopic characterization, but is apparently negatively charged and retains the two chlorine atoms of the substrate. The enzyme was partially sequenced using electrospray LC-MS, and one peptide was used to search the NCBInr database. This peptide matched a part of PcpA, a protein of unknown function that is induced in S. chlorophenolica in response to pentachlorophenol. Several other peptides could also be mapped onto the sequence of PcpA, suggesting that the enzyme is encoded by pcpA. PcpA has low but significant sequence similarity to an unusual class of extradiol dioxygenases. On the basis of the sequence analysis, the Fe2+ and O2 dependence of the enzyme, and the characteristics of the product, the enzyme is proposed to be a 2,6-dichlorohydroquinone dioxygenase. The position of ring cleavage has not yet been identified. PMID- 10387006 TI - The role of beta-Arg-182, an essential catalytic site residue in Escherichia coli F1-ATPase. AB - Beta-Arg-182 in Escherichia coli F1-ATPase (beta-Arg-189 in bovine mitochondrial F1) is a residue which lies close to catalytic site bound nucleotide (Abrahams et al. (1994) Nature 370, 621-628). Here we investigated the role of this residue by characterizing two mutants, betaR182Q and betaR182K. Oxidative phosphorylation and steady-state ATPase activity of purified F1 were severely impaired by both mutations. Catalytic site nucleotide-binding parameters were measured using the fluorescence quench of beta-Trp-331 that occurred upon nucleotide binding to purified F1 from betaR182Q/betaY331W and betaR182K/betaY331W double mutants. It was found that (a) beta-Arg-182 interacts with the gamma-phosphate of MgATP, particularly at catalytic sites 1 and 2, (b) beta-Arg-182 has no functional interaction with the beta-phosphate of MgADP or with the magnesium of the magnesium-nucleotide complex in the catalytic sites, and (c) beta-Arg-182 is directly involved in the stabilization of the catalytic transition state. In these features the role of beta-Arg-182 resembles that of another positively charged residue in the catalytic site, the conserved lysine of the Walker A motif, beta-Lys-155. A further role of beta-Arg-182 is suggested, namely involvement in conformational change at the catalytic site beta-alpha subunit interface that is required for multisite catalysis. PMID- 10387007 TI - Escherichia coli methionine aminopeptidase: implications of crystallographic analyses of the native, mutant, and inhibited enzymes for the mechanism of catalysis. AB - By improving the expression and purification of Escherichia coli methionine aminopeptidase (eMetAP) and using slightly different crystallization conditions, the resolution of the parent structure was extended from 2.4 to 1.9 A resolution. This has permitted visualization of the coordination geometry and solvent structure of the active-site dinuclear metal center. One solvent molecule (likely a mu-hydroxide) bridges the trigonal bipyramidal (Co1) and octahedral (Co2) cobalt ions. A second solvent (possibly a hydroxide ion) is bound terminally to Co2. A monovalent cation binding site was also identified about 13 A away from the metal center at an interface between the two subdomains of the protein. The first structure of a substrate-like inhibitor, (3R)-amino-(2S)-hydroxyheptanoyl-L Ala-L-Leu-L-Val-L-Phe-OMe, bound to a methionine aminopeptidase, has also been determined. This inhibitor coordinates the metal center through four interactions as follows: (i) ligation of the N-terminal (3R)-nitrogen to Co2, (ii, iii) bridging coordination of the (2S)-hydroxyl group, and (iv) terminal ligation to Co1 by the keto oxygen of the pseudo-peptide linkage. Inhibitor binding occurs with the displacement of two solvent ligands and the expansion of the coordination sphere of Co1. In addition to the tetradentate, bis-chelate metal coordination, the substrate analogue forms hydrogen bonds with His79 and His178, two conserved residues within the active site of all MetAPs. To evaluate their importance in catalysis His79 and His178 were replaced with alanine. Both substitutions, but especially that of His79, reduce activity. The structure of the His79Ala apoenzyme and the comparison of its electronic absorption spectra with other variants suggest that the loss in activity is not due to a conformational change or a defective metal center. Two different reaction mechanisms are proposed and are compared to those of related enzymes. These results also suggest that inhibitors analogous to that reported here may be useful in preventing angiogenesis in cancer and in the treatment of microbial and fungal infections. PMID- 10387008 TI - Circadian synthesis of a nuclear-encoded chloroplast glyceraldehyde-3-phosphate dehydrogenase in the dinoflagellate Gonyaulax polyedra is translationally controlled. AB - The circadian clock has previously been shown to restrict synthesis of several proteins in the dinoflagellate Gonyaulax polyedra to only a few hours each day. We have identified one of these proteins as glyceraldehyde-3-phosphate dehydrogenase. Two nuclear genes encoding the enzyme have been cloned, one corresponding to a cytoplasmic isoform and the other to a plastid targeted protein. On the basis of protein microsequence data, we conclude that the synthesis of the plastid isoform is clock-regulated. This regulation is not related to mRNA levels, which remain constant throughout the cycle, suggesting a translational control mechanism, in contrast to the transcriptional regulation of GAPDH that has been demonstrated in Neurospora. Although the rhythm of synthesis has a high amplitude, the abundance and activity rhythms are greatly attenuated, which is attributed to the long half-life of the protein. PMID- 10387009 TI - Sliding clamp of the bacteriophage T4 polymerase has open and closed subunit interfaces in solution. AB - The sliding clamps of bacteriophage T4 (gp45), Escherichia coli (beta clamp), and yeast (PCNA) are required for processive DNA synthesis by their cognate DNA polymerases. The X-ray crystal structures of all three of these clamps have been shown to be closed, circular complexes. This paper reports investigations of the solution structure of bacteriophage T4 gp45 by analytical ultracentrifugation, fluorescence, and hydrodynamic modeling. Mutants of gp45 with inter- and intrasubunit disulfide bonds were created to alter the solution structure of gp45, with additional mutagenesis used to investigate the importance of the proline-rich loop region found between the two domains of each gp45 monomer. The wild-type gp45 trimer assembles from monomers cooperatively with a dissociation constant of 0.21 microM2 and values between 0.088 and 0. 32 microM2 for the mutants. Velocity ultracentrifugation experiments showed that wild-type gp45 possesses a sedimentation coefficient strongly dependent on concentration, typical of asymmetric or elongated molecules, that when extrapolated to zero concentration yields a sedimentation coefficient of 4.0 S. The loop and the disulfide mutants exhibited sedimentation coefficients with little concentration dependence, typical of symmetric or spherical molecules, that when extrapolated to zero concentration yielded sedimentation coefficients of 4.4-4.8 S. The lower sedimentation coefficient in the former case is consistent with wild-type gp45 being more asymmetric or elongated than the mutant forms. Fluorescence-resonance energy-transfer experiments were used to measure the distance between two amino acids (W91 and V162C-coumarin) on opposite sides of the gp45 subunit interface. For an intrasubunit disulfide mutant, the distance between these two amino acids was determined to be 19 A (14 A in the X-ray crystal structure), consistent with a closed complex. For the mutants without intrasubunit disulfides, the efficiency of fluorescence-resonance energy transfer was in accord with a model of gp45 being an open complex composed of two closed subunit interfaces and a third open interface separated by a distance of 35-38 A. The collective data supplemented with hydrodynamic modeling were consistent with gp45 subunit separation achieved within the plane of the gp45 ring. PMID- 10387010 TI - Critical nature of a specific uridine O2-carbonyl for cleavage by the hammerhead ribozyme. AB - Three modified hammerhead ribozyme/substrate complexes have been prepared in which individual uridine O2-carbonyls have been eliminated. The modified complexes were chemically synthesized with the substitution of a single 2 pyridone (2P) base analogue for residues U4, U7, and U16.1. Steady-state kinetic analyses indicate that the cleavage efficiencies for the U7 and U16.1 complexes were not significantly reduced relative to the native complex as measured by kcat/KM. The cleavage efficiency for the 2P4 complex, with the analogue present within the uridine loop, was reduced by greater than 2 orders of magnitude. This significant reduction in catalytic efficiency was due primarily to a decrease in kcat. The pH vs cleavage rate profile suggests that the O2-carbonyl of the U4 residue of the hammerhead complex is critical for transition state stabilization and efficient cleavage activity. The results of a Mg2+ rescue assay do not implicate the O2-carbonyl of U4 in an interaction with a divalent metal ion. In addition, the results of a ribozyme folding assay suggest that the presence of the 2P4 within the uridine loop does not alter the folding pathway (relative to the native sequence) both in the absence and in the presence of Mg2+. The O2 carbonyl of U4 appears oriented toward the interior of the catalytic pocket where it may be involved in a critical hydrogen bonding interaction necessary for transition state stabilization. PMID- 10387012 TI - Arg304 of human DNA primase is a key contributor to catalysis and NTP binding: primase and the family X polymerases share significant sequence homology. AB - Comparison of the amino acid sequences of eucaryotic DNA primase and the family X polymerases indicates that primase shares significant sequence homology with this family. With the use of DNA polymerase beta (pol beta) as a paradigm for family X polymerases, these homologies include both the catalytic core domain/subunit of each enzyme (31 kDa domain of pol beta and p49 subunit of primase) as well as the accessory domain/subunit (8 kDa domain of pol beta and p58 subunit of primase). To further explore these homologies as well as provide insights into the mechanism of primase, we generated three mutants (R304K, R304Q, and R304A) of the p49 subunit at an arginine that is highly conserved between primase and the eukaryotic family X polymerases. These mutations significantly decreased the rate of primer synthesis, due primarily to a decreased rate of initiation, and the extent of impairment correlated with the severity of the mutation (A > Q > K). R304 also contributes to efficient utilization of the NTP that will become the 5' terminus of the new primer, and these effects are at least partially mediated through interactions with the phosphates of this NTP. The implications of these results with respect to the structure and biological role of primase, as well as its relationship to the family X polymerases, are discussed. PMID- 10387011 TI - Stimulation of topoisomerase II-mediated DNA cleavage by three DNA-intercalating plant alkaloids: cryptolepine, matadine, and serpentine. AB - Cryptolepine, matadine, and serpentine are three indoloquinoline alkaloids isolated from the roots of African plants: Cryptolepis sanguinolenta, Strychnos gossweileri, and Rauwolfia serpentina, respectively. For a long time, these alkaloids have been used in African folk medicine in the form of plant extracts for the treatment of multiple diseases, in particular as antimalarial drugs. To date, the molecular basis for their diverse biological effects remains poorly understood. To elucidate their mechanism of action, we studied their interaction with DNA and their effects on topoisomerase II. The strength and mode of binding to DNA of the three alkaloids were investigated by spectroscopy. The alkaloids bind tightly to DNA and behave as typical intercalating agents. All three compounds stabilize the topoisomerase II-DNA covalent complex and stimulate the cutting of DNA by topoisomerase II. The poisoning effect is more pronounced with cryptolepine than with matadine and serpentine, but none of the drugs exhibit a preference for cutting at a specific base. Cryptolepine which binds 10-fold more tightly to DNA than the two related alkaloids proves to be much more cytotoxic toward B16 melanoma cells than matadine and serpentine. The cellular consequences of the inhibition of topoisomerase II by cryptolepine were investigated using the HL60 leukemia cell line. The flow cytometry analysis shows that the drug alters the cell cycle distribution, but no sign of drug-induced apoptosis was detected when evaluating the internucleosomal fragmentation of DNA in cells. Cryptolepine treated cells probably die via necrosis rather than via apoptosis. The results provide evidence that DNA and topoisomerase II are the primary targets of cryptolepine, matadine, and serpentine. PMID- 10387013 TI - Synthetase recognition determinants of E. coli valine transfer RNA. AB - We have studied the interactions between Escherichia coli tRNAVal and valyl-tRNA synthetase (ValRS) by enzymatic footprinting with nuclease S1 and ribonuclease V1, and by analysis of the aminoacylation kinetics of mutant tRNAVal transcripts. Valyl-tRNA synthetase specifically protects the anticodon loop, the 3' side of the stacked T-stem/acceptor-stem helix, and the 5' side of the anticodon stem of tRNAVal against cleavage by double- and single-strand-specific nucleases. Increased nuclease susceptibility at the ends of the anticodon- and T-stems in the tRNAVal.ValRS complex is indicative of enzyme-induced conformational changes in the tRNA. The most important synthetase recognition determinants are the middle and 3' anticodon nucleotides (A35 and C36, respectively); G20, in the variable pocket, and G45, in the tRNA central core, are minor recognition elements. The discriminator base, position 73, and the anticodon stem also are recognized by ValRS. Replacing wild-type A73 with G73 reduces the aminoacylation efficiency more than 40-fold. However, the C73 and U73 mutants remain good substrates for ValRS, suggesting that guanosine at position 73 acts as a negative determinant. The amino acid acceptor arm of tRNAVal contains no other synthetase recognition nucleotides, but regular A-type RNA helix geometry in the acceptor stem is essential [Liu, M., et al. (1997) Nucleic Acids Res. 25, 4883-4890]. In the anticodon stem, converting the U29:A41 base pair to C29:G41 reduces the aminoacylation efficiency 50-fold. This is apparently due to the rigidity of the anticodon stem caused by the presence of five consecutive C:G base pairs, since the A29:U41 mutant is readily aminoacylated. Identity switch experiments provide additional evidence for a role of the anticodon stem in synthetase recognition. The valine recognition determinants, A35, C36, A73, G20, G45, and a regular A-RNA acceptor helix are insufficient to transform E. coli tRNAPhe into an effective valine acceptor. Replacing the anticodon stem of tRNAPhe with that of tRNAVal, however, converts the tRNA into a good substrate for ValRS. These experiments confirm G45 as a minor ValRS recognition element. PMID- 10387014 TI - Influence of the physical state of the membrane on the enzymatic activity and energy of activation of protein kinase C alpha. AB - The activation of protein kinase C alpha was studied by using a lipid system consisting of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)/1-palmitoyl 2-oleoyl-sn-glycero-3-phosphoserine (POPS) (molar ratio 4:1) and different proportions of 1-palmitoyl-2-oleoyl-sn-glycerol (POG). The phase behavior of the lipidic system was characterized by using differential scanning calorimetry and 31P NMR, and a phase diagram was elaborated. The results suggested the formation of two diacylglycerol/phospholipid complexes, one at 15 mol % of POG and the second at 30 mol % of POG. These two complexes would define the three regions of the phase diagram: in the first region (concentrations of POG lower than 15 mol %) there is gel-gel immiscibility at temperatures below that of the phase transition between C1 and pure phospholipid, and a fluid lamellar phase above of the phase transition. In the second region (between 15 and 30 mol % of POG), gel gel immiscibility between C1 and C2 with fluid-fluid immiscibility was observed, while inverted hexagonal HII and isotropic phases were detected by 31P NMR. In the third region (concentrations of POG higher than 30 mol %), gel-gel immiscibility seemed to occur between C2 and pure POG along with fluid-fluid immiscibility, while an isotropic phase was detected by 31P NMR. When PKC alpha activity was measured, as a function of POG concentration, maximum activity was found at POG concentrations as low as 5-10 mol %; the activity slightly decreased as POG concentration was increased to 45 mol % at 32 degrees C (above Tc) whereas activity did not change with increasing concentrations of POG at 5 degrees C (below Tc). When the activity was studied as a function of temperature, at different POG concentrations, and depicted as Arrhenius plots, it was found that the activity increased with increasing temperatures, showing a discontinuity at a temperature very close to the phase transition of the system and a lower activation energy at the upper slope of the graph, indicating that the physical state of the membrane affected the interaction of PKC alpha with the membrane. PMID- 10387015 TI - Suppression of GTP/T alpha-dependent activation of cGMP phosphodiesterase by ADP ribosylation by its gamma subunit in amphibian rod photoreceptor membranes. AB - Our previous study has shown that P gamma, the regulatory subunit of cGMP phosphodiesterase (PDE), is ADP-ribosylated by endogenous ADP-ribosyltransferase when P gamma is free or complexed with the catalytic subunits of PDE in amphibian rod photoreceptor membranes. The P gamma domain containing ADP-ribosylated arginines was shown to be involved in its interaction with T alpha, a key interaction for PDE activation. In this study, we describe a possible function of the P gamma ADP-ribosylation in the GTP/T alpha-dependent PDE activation. When rod membranes were preincubated with or without NAD and washed with a buffer containing GTP, the PDE activity of NAD-preincubated membranes was increased by the GTP-washing only to approximately 50% of that of membranes preincubated without NAD. The P gamma release by the GTP-washing from these NAD-preincubated membranes was also suppressed to approximately 50% of that preincubated without NAD. Taking into consideration that approximately 50% of P gamma is ADP ribosylated under these conditions, these observations suggest that the ADP ribosylated P gamma cannot interact with GTP/T alpha. We have also shown that a soluble fraction of ROS contains an enzyme(s) to release the radioactivity of [32P]ADP-ribosylated P gamma in concentration- and time-dependent manners, suggesting that the P gamma ADP-ribosylation is reversible. Rod ADP ribosyltransferase solubilized from membranes by phosphatidylinositol-specific phospholipase C was separated into two fractions by ion-exchange columns. Biochemical characterization of these two fractions, including measurement of the Km for NAD and P gamma, estimation of their molecular masses, ADP-ribosylation of P gamma arginine mutants, effects of ADP-ribosyltransferase inhibitors on the P gamma ADP-ribosylation, and effects of salts and pH on the P gamma ADP ribosylation, indicates that rod ADP-ribosyltransferase contains two isozymes, and that these two isozymes have similar properties for the P gamma ADP ribosylation. Our observations strongly suggest that the negative regulation of PDE through the reversible P gamma ADP-ribosylation may function in the phototransduction mechanism. PMID- 10387016 TI - Biochemical analysis of the Saccharomyces cerevisiae SEC18 gene product: implications for the molecular mechanism of membrane fusion. AB - The SEC18 gene product is 48% identical to mammalian NSF (N-ethylmaleimide sensitive fusion protein), and both proteins encode cytoplasmic ATPases which are essential for membrane traffic in yeast and mammalian cells, respectively. A wealth of biochemical analysis has led to the description of a model for the action of NSF; through its interaction with SNAPs (soluble NSF attachment proteins), NSF can associate with SNAP receptors (SNAREs) on intracellular membranes, forming 20S complexes. SNAPs then stimulate the intrinsic ATPase activity of NSF, leading to the disassembly of the 20S complex, which is essential for subsequent membrane fusion. Although this model is based almost entirely on in vitro studies of the original clones of NSF and alpha-SNAP, it is nevertheless widely assumed that this mechanism of membrane fusion is conserved in all eukaryotic cells. If so, the crucial biochemical properties of NSF and SNAPs should be shared by their yeast homologues, Sec18p and Sec17p. Using purified recombinant proteins, we report here that Sec18p can specifically interact not only with Sec17p but also with its mammalian homologue, alpha-SNAP. This interaction leads to a stimulation of Sec18p D1 domain ATPase activity, with kinetics similar to those of alpha-SNAP stimulation of NSF, although differences in temperature and N-ethylmaleimide sensitivity were observed between NSF and Sec18p. Furthermore, Sec18p can interact with synaptic SNARE proteins and can synergize with alpha-SNAP to stimulate regulated exocytosis in mammalian cells. We conclude that the mechanistic properties of NSF and SNAPs are shared by Sec18p and Sec17p, thus demonstrating that the biochemistry of membrane fusion is conserved from yeast to mammals. PMID- 10387017 TI - Modulation of the affinity and selectivity of RGS protein interaction with G alpha subunits by a conserved asparagine/serine residue. AB - The crystal structure of the complex between a G protein alpha subunit (Gi alpha 1) and its GTPase-activating protein (RGS4) demonstrated that RGS4 acts predominantly by stabilization of the transition state for GTP hydrolysis [Tesmer, J. J., et al. (1997) Cell 89, 251]. However, attention was called to a conserved Asn residue (Asn128) that could play a catalytic role by interacting, directly or indirectly, with the hydrolytic water molecule. We have analyzed the effects of several disparate substitutions for Asn128 on the GAP activity of RGS4 toward four G alpha substrates (Go, Gi, Gq, and Gz) using two assay formats. The results substantiate the importance of this residue but indicate that it is largely involved in substrate binding and that its function may vary with different G alpha targets. Various mutations decreased the apparent affinity of RGS4 for substrate G alpha proteins by several orders of magnitude, but had variable and modest effects on maximal rates of GTP hydrolysis when tested with different G alpha subunits. One mutation, N128F, that differentially decreased the GAP activity toward G alpha i compared with that toward G alpha q could be partially suppressed by mutation of the nearby residue in G alpha i to that found in G alpha q (K180P). Detection of GAP activities of the mutants was enhanced in sensitivity up to 100-fold by assay at steady state in proteoliposomes that contain heterotrimeric G protein and receptor. PMID- 10387018 TI - Comparison of in vivo and in vitro phosphorylation of the exocytosis-sensitive protein PP63/parafusin by differential MALDI mass spectrometric peptide mapping. AB - PP63 (parafusin) is a 63 kDa phosphoprotein, which exists in at least two different isoforms. It is very rapidly (80 ms) dephosphorylated during triggered trichocyst exocytosis. This occurs selectively in exocytosis-competent Paramecium tetraurelia strains. At least two protein kinases isolated from Paramecium, casein kinase type II kinase and cGMP-dependent kinase, are able to phosphorylate the two recombinant PP63/parafusin isoforms, both with phosphoglucomutase activity, in vitro. By performing mass spectrometric peptide mapping, we have investigated in vitro phosphorylation of recombinant PP63/parafusin by these kinases in comparison to in vivo phosphorylation of native PP63/parafusin isolated from Paramecium homogenates. Low picomolar quantities of proteolytic digests of recombinant and native PP63/parafusin, prior to and following alkaline phosphatase treatment, were directly analyzed by MALDI mass spectrometry. In native PP63-1/parafusin-1, six of 64 serine and threonine residues (S-196, T-205, T-280, T-371, T-373, and T-469) were found definitely, 27 were found possibly phosphorylated, 28 were identified as nonphosphorylated, and three were not covered by mapping. Three of the six certainly phosphorylated amino acids represent consensus phosphorylation sites for casein kinase II or cGMP-dependent protein kinase. In vitro phosphorylation studies of recombinant PP63/parafusin confirm that some of the sites found were used in vivo; however, also significant differences with respect to in vivo phosphorylation of native PP63/parafusin were observed. The two Paramecium protein kinases that were used do not preferably phosphorylate expected consensus sites in vitro. Homology structure modeling of PP63/parafusin with rabbit phosphoglucomutase revealed that the majority of residues found phosphorylated is located on the surface of the molecule. PMID- 10387019 TI - Stages in iron storage in the ferritin of Escherichia coli (EcFtnA): analysis of Mossbauer spectra reveals a new intermediate. AB - Iron uptake into the nonheme ferritin of Escherichia coli (EcFtnA) and its site directed variants have been investigated by Mossbauer spectroscopy. EcFtnA, like recombinant human H chain ferritin (HuHF), oxidized Fe(II) at a dinuclear ferroxidase center situated at a central position within each subunit. As with HuHF, Mossbauer subspectra observed between 1 min and 24 h after Fe(II) addition were assigned to Fe(III) monomers, "c", mu-oxo-bridged dimers, "b", and clusters, "a", the latter showing magnetically split spectra, "d", at 4.1 K. Like those of HuHF, the mu-oxo-bridged dimers were formed at the ferroxidase centers. However, the analysis also revealed the presence of a new type of dimer, "e" (QS1 = 0.38 mm/s, IS1 = 0.51 mm/s and QS2 = 0.72 mm/s, IS2 = 0.50 mm/s), and this was also assigned to the ferroxidase center. Dimers "b" appeared to be converted to dimers "e" over time. Subspectra "e" became markedly asymmetric at temperatures above 90 K, suggesting that the two Fe(III) atoms of dimers "e" were more weakly coupled than in the mu-oxo-bridged dimers "b", possibly due to OH- bridging. Monomeric Fe(III), giving relaxation spectra "c", was assigned to a unique site C that is near the dinuclear center. In EcFtnA all three iron atoms seemed to be oxidized together. In contrast to HuHF, no Fe(III) clusters were observed 24 h after the aerobic addition of 48 Fe(II) atoms/molecule in wild-type EcFtnA. This implies that iron is more evenly distributed between molecules in the bacterial ferritins, which may account for its greater accessibility. PMID- 10387020 TI - Differential interfacial and substrate binding modes of mammalian pancreatic phospholipases A2: a comparison among human, bovine, and porcine enzymes. AB - To identify the residues essential for interfacial binding and substrate binding of human pancreatic phospholipase A2 (hpPLA2), several ionic residues in the putative interfacial binding surface (R6E, K7E, K10E, and K116E) and substrate binding site (D53K and K56E) were mutated. Interfacial affinity of these mutants was measured using anionic polymerized liposomes, and their enzymatic activity was measured using various substrates including phospholipid monomers, zwitterionic and anionic micelles, and anionic polymerized mixed liposomes. Similar mutations (R6E, K10E, K56E, and K116E) were made to porcine pancreatic phospholipase A2 (ppPLA2), and the properties of mutants were measured by the same methods. Results indicate that hpPLA2 and ppPLA2 have similar interfacial binding mechanisms in which cationic residues in the amino terminus and Lys-116 in the carboxy terminus are involved in binding to anionic lipid surfaces. Small but definite differences between the two enzymes were observed in overall interfacial affinity and activity and the effects of the mutations on interfacial enzyme activity. The interfacial binding of hpPLA2 and ppPLA2 is distinct from that of bovine pancreatic phospholipase A2 in that Lys-56 is involved in the interfacial binding of the latter enzyme. The unique phospholipid headgroup specificity of hpPLA2 derives from the presence of Asp-53 in the substrate binding site. This residue appears to participate in stabilizing electrostatic interactions with the cationic ethanolamine headgroup, hence the phosphatidylethanolamine preference of hpPLA2. Taken together, these studies reveal the similarities and the differences in the mechanisms by which mammalian pancreatic phospholipases A2 interact with lipid aggregates and perform interfacial catalysis. PMID- 10387022 TI - Role of the distal phenylalanine 54 on the structure, stability, and ligand binding of Coprinus cinereus peroxidase. AB - Resonance Raman and electronic absorption spectra obtained at various pH values for the Fe3+ form of distal F54 mutants of Coprinus cinereus peroxidase are reported, together with the Fe2+ form and fluoride and imidazole adducts at pH 6.0, 5.0, and 10.5, respectively. The distal phenylalanine residue has been replaced by the small aliphatic residues glycine and valine and the hydrogen bonding aromatic residues tyrosine and tryptophan (F54G, -V, -Y, and -W, respectively). These mutations resulted in transitions between ferric high-spin five-coordinate and six-coordinate forms, and caused a decrease of the pKa of the alkaline transition together with a higher tendency for unfolding. The mutations also alter the ability of the proteins to bind fluoride in such a way that those that are six-coordinate at pH 5.0 bind more strongly than both wild-type CIP and F54Y which are five-coordinate at this pH value. The data provide evidence that the architecture of the distal pocket of CIP is altered by the mutations. Direct evidence is provided that the distal phenylalanine plays an important role in controlling the conjugation between the vinyl double bonds and the porphyrin macrocycle, as indicated by the reorientation of the vinyl groups upon mutation of phenylalanine with the small aliphatic side chains of glycine and valine residues. Furthermore, it appears that the presence of the hydrogen-bonding tyrosine or tryptophan in the cavity increases the pKa of the distal histidine for protonation compared with that of wild-type CIP. PMID- 10387021 TI - A structural determinant of the unique interfacial binding mode of bovine pancreatic phospholipase A2. AB - The catalytic steps of the phospholipase A2 (PLA2)-catalyzed hydrolysis of phospholipids are preceded by interfacial binding. Among various pancreatic PLA2s, bovine pancreatic PLA2 (bpPLA2) has a unique interfacial binding mode in which Lys-56 plays an important role in its binding to anionic lipid surfaces. To identify the structural determinant of this unique interfacial binding mode of bpPLA2, we systematically mutated bpPLA2 and measured the effects of mutations on its interfacial binding and activity. First, different cationic clusters were generated in the amino-terminal alpha-helix by the N6R, G7K, and N6R/G7K mutations. These mutations enhanced the binding of bpPLA2 to anionic liposomes up to 15-fold. For these mutants, however, the K56E mutation still caused a large drop in interfacial affinity for and activity toward anionic liposomes, indicating that the generation of a cationic patch in the amino-terminal alpha helix of bpPLA2 did not change its interfacial binding mode. Second, residues 62 66 that form a part of the pancreatic loop were deleted. For this deletion mutant (Delta62-66), which was as active as wild-type toward anionic liposomes, the K56E and K116E mutations (Delta62-66/K56E and Delta62-66/K116E) did not have significant effects on interfacial affinity. In contrast, the K10E mutation showed a much larger decrease in interfacial affinity (10-fold), indicating the deletion of residues 62-66 caused a major change in the interfacial binding mode. Finally, hydrophobic residues in positions 63 and 65 were replaced by bulkier ones (V63F and V63F/V65L) to pinpoint the structural determinant of the interfacial binding mode of bpPLA2. The effects of K10E and K56E mutations on the interfacial affinity and activity of these mutants showed that Val-63 and Val-65 of bpPLA2 are the structural determinant of its unique interfacial binding mode and that relatively conservative substitutions at these positions result in large changes in the interfacial binding mode among mammalian pancreatic PLA2s. Taken together, this study reveals how minor structural differences among homologous PLA2s can lead to distinct interfacial binding behaviors. PMID- 10387023 TI - Comparison of the substrate specificities of human liver cytochrome P450s 2C9 and 2C18: application to the design of a specific substrate of CYP 2C18. AB - A series of 2-aroylthiophenes derived from tienilic acid by replacement of its OCH2COOH substituent with groups bearing various functions have been synthesized and studied as possible substrates of recombinant human liver cytochrome P450s 2C9 and 2C18 expressed in yeast. Whereas only compounds bearing a negative charge acted as substrates of CYP 2C9 and were hydroxylated at position 5 of their thiophene ring at a significant rate, many neutral 2-aroylthiophenes were 5 hydroxylated by CYP 2C18 with kcat values of >2 min-1. Among the various compounds that were studied, those bearing an alcohol function were the best CYP 2C18 substrates. One of them, compound 3, which bears a terminal O(CH2)3OH function, appeared to be a particularly good substrate of CYP 2C18. It was regioselectively hydroxylated by CYP 2C18 at position 5 of its thiophene ring with a KM value of 9 +/- 1 microM and a kcat value of 125 +/- 25 min-1, which are the highest described so far for a CYP 2C. A comparison of the oxidations of 3, by yeast-expressed CYP 1A1, 1A2, 2C8, 2C9, 2C18, 2C19, 2D6, 2E1, 3A4, and 3A5, showed that only CYP 2C8, 2C18, and 2C19 were able to catalyze the 5 hydroxylation of 3. However, the catalytic efficiency of CYP 2C18 for that reaction was considerably higher (kcat/KM value being 3-4 orders of magnitude larger than those found for CYP 2C8 and 2C19). Several human P450s exhibited small activities for the oxidative O-dealkylation of 3. The four recombinant CYP 2Cs were the best catalysts for that reaction (kcat between 1 and 5 min-1) when compared to all the P450s that were tested, even though it is a minor reaction in the case of CYP 2C18. All these results show that compound 3 is a new, selective, and highly efficient substrate for CYP 2C18 that should be useful for the study of this P450 in various organs and tissues. They also suggest some key differences between the active sites of CYP 2C9 and CYP 2C18 for substrate recognition. PMID- 10387024 TI - On the mechanism of inhibition of the nicotinic acetylcholine receptor by the anticonvulsant MK-801 investigated by laser-pulse photolysis in the microsecond to-millisecond time region. AB - The mechanism of inhibition of the muscle nicotinic acetylcholine receptor is of interest because of the many drugs which are known to modify its function. The laser-pulse photolysis technique, using a photolabile, biologically inert ligand (caged carbamoylcholine) for the nicotinic acetylcholine receptor, and BC3H1 cells have been used to investigate the mechanism of inhibition of the receptor by MK-801 [(+)-dizocilpine] in the microsecond-to-millisecond time region. MK-801 is an anticonvulsant and a known inhibitor of the N-methyl-D-aspartate and nicotinic acetylcholine receptors. Both the chemical kinetic and the single channel current-recording measurements reported here indicate the existence of two inhibition processes, one occurring within 50 ms and the other within about 1 s of equilibration of the receptor with the inhibitor. Unless stated otherwise, here we characterize the receptor inhibition observed when MK-801 is equilibrated with the receptor for only 50 ms. We determined the effect of MK-801 on the concentration of the open receptor-channels and the apparent dissociation constant of the inhibitor from the closed-channel (KI(obs) = 180 microM) and open channel ( = 950 microM) forms. Within a few milliseconds after inhibitor binding, decreases to about 100 microM, due to an inhibitor-induced isomerization to an inactive receptor form. A mechanism that incorporates the new results is proposed. It includes the formation of an ion-conducting receptor:inhibitor complex with a channel-opening equilibrium constant that is unfavorable compared to the open-channel receptor form in the absence of inhibitor. In the MK-801 concentration range of 0-500 microM, this mechanism accounts for the observed MK 801-induced decrease in the concentration of open channels. At high concentrations of carbamoylcholine, when the receptor is mainly in the open channel form, the conducting receptor:inhibitor complex isomerizes to a nonconducting state with a rate constant of about 2400 s-1 for the forward reaction and 230 s-1 for the back reaction. It is shown that the proposed new mechanism, based on transient kinetic measurements, also accounts for the results of previous investigations with other inhibitors (procaine, cocaine), which were carried out under both pre-steady-state and equilibrium conditions. A compound that binds to the same regulatory site on the receptor as MK-801 but does not affect the channel-opening equilibrium constant may have considerable use in protecting an organism from the effects of abused drugs. PMID- 10387025 TI - The functional role of positively charged amino acid side chains in alpha bungarotoxin revealed by site-directed mutagenesis of a His-tagged recombinant alpha-bungarotoxin. AB - A polyhistidine tag was added to the N-terminus of alpha-bungarotoxin (Bgtx) recombinantly expressed in E. coli. The His-tagged Bgtx was identical to native, venom-derived Bgtx in its apparent affinity for the nicotinic acetylcholine receptor (nAChR) in Torpedo electric organ membranes. Furthermore, in a physiological assay involving mouse muscle nAChR expressed in Xenopus oocytes, the His-tagged Bgtx was as effective as authentic Bgtx at blocking acetylcholine evoked currents. Ala-substitution mutagenesis of His-tagged Bgtx was used to evaluate the functional contribution of Arg36, a residue that is invariant among all alpha-neurotoxins. Replacement with Ala resulted in a 90-fold decrease in the apparent affinity for the Torpedo nAChR and a corresponding 150-fold increase in the IC50 for block of heterologously expressed mouse muscle nAChR, demonstrating the critical importance of this positive charge for the binding and functional activity of a long alpha-neurotoxin. The observed decrease in affinity corresponds to a DeltaDeltaG of 2.7 kcal/mol and indicates that Arg36 makes a major contribution to complex formation. This finding is consistent with the proposal that Arg36 mimics the positive charge found on acetylcholine and directs the toxin to interact with receptor sites normally involved in acetylcholine recognition. In comparison, Ala-substitution of the highly conserved Lys26 resulted in only a 9-fold decrease in apparent affinity. Truncation of the His tagged Bgtx following residue 67 produces a toxin lacking the seven C-terminal residues including the two positively charged residues Lys70 and Arg72. Truncation leads to a 7-fold decrease in apparent binding affinity. PMID- 10387026 TI - The arginine 276 anchor for NADP(H) dictates fluorescence kinetic transients in 3 alpha-hydroxysteroid dehydrogenase, a representative aldo-keto reductase. AB - Fluorescence stopped-flow studies were conducted with recombinant rat liver 3 alpha-HSD, an aldo-keto reductase (AKR) that plays critical roles in steroid hormone inactivation, to characterize the binding of nicotinamide cofactor, the first step in the kinetic mechanism. Binding of NADP(H) involved two events: the fast formation of a loose complex (E.NADP(H)), followed by a conformational change in enzyme structure leading to a tightly bound complex (E.NADP(H)), which was observed as a fluorescence kinetic transient. Binding of NAD(H) was not characterized by a similar kinetic transient, implying a difference in the mode of binding of the two cofactors. Unlike previously characterized AKRs, the rates associated with the formation and decay of E.NADP(H) and E.NADP(H) were much faster than kcat for the oxidoreduction of various substrates, indicating that binding and release of cofactor is not rate-limiting overall in 3 alpha-HSD. Mutation of Arg 276, a highly conserved residue in AKRs that forms a salt bridge with the adenosine 2'-phosphate of NADP(H), resulted in large changes in Km and Kd for NADP(H) that were not observed with NAD(H). The loss in free energy associated with the increase in Kd for NADP(H) is consistent with the elimination of an electrostatic link. Importantly, this mutation abolished the kinetic transient associated with NADPH binding. Thus, anchoring of the adenosine 2' phosphate of NADPH by Arg 276 appears to be obligatory for the fluorescence kinetic transients to be observed. The removal of Trp 86, a residue involved in fluorescence energy transfer with NAD(P)H, also abolished the kinetic transient, but mutation of Trp 227, a residue on a mobile loop associated with cofactor binding, did not. It is concluded that in 3 alpha-HSD, the time dependence of the change in Trp 86 fluorescence is due to cofactor anchoring, and thus, Trp 86 is a distal reporter of this event. Further, the loop movement that accompanies cofactor binding is spectrally silent. PMID- 10387027 TI - Thermal denaturation of human gamma-interferon. A calorimetric and spectroscopic study. AB - The thermal denaturation of a recombinant human gamma-interferon has been studied as a function of pH in the range from 2 to 10 and buffer concentration in the range from 5 to 100 mM by differential scanning calorimetry, circular dichroism, fluorescence, 1H NMR, and biological activity measurements. The thermal transitions are irreversible at high buffer concentrations at all pH values studied, although they are reversible between pH 3.5 and 5.4 at low buffer concentrations. The denaturation enthalpy, DeltaH(Tm), at denaturation temperature Tm was a function of both Tm and the buffer concentration, and this resulted in heat capacity changes decreasing with buffer concentration. When the denaturation enthalpies were corrected for Tm dependence, they did not appear to change versus pH. The denaturation entropies, however, appeared to decrease with pH, leading to a small but appreciable increase in the stability of the protein with pH. The difference between the number of moles of protons stoichiometrically bound to a mole of protein in the native and thermally denatured state, was calculated from the variation of Tm versus pH at each buffer concentration. The values obtained appear to depend on pH alone rather than upon temperature or buffer concentration, a result which agrees with the invariance of the denaturation enthalpies with pH. This dependence was fitted to the titration curve of a group with a pK of 5.4. PMID- 10387028 TI - Redox properties of mesophilic and hyperthermophilic rubredoxins as a function of pressure and temperature. AB - The formal equilibrium reduction potentials of recombinant electron transport protein, rubredoxin (MW = 7500 Da), from both the mesophilic Clostridium pasteurianum (Topt = 37 degrees C) and hyperthermophilic Pyrococcus furiosus (Topt = 95 degrees C) were recorded as a function of pressure and temperature. Measurements were made utilizing a specially designed stainless steel electrochemical cell that easily maintains pressures between 1 and 600 atm and a temperature-controlled cell that maintains temperatures between 4 and 100 degrees C. The reduction potential of P. furiosus rubredoxin was determined to be 31 mV at 25 degrees C and 1 atm, -93 mV at 95 degrees C and 1 atm, and 44 mV at 25 degrees C and 400 atm. Thus, the reduction potential of P. furiosus rubredoxin obtained under standard conditions is likely to be dramatically different from the reduction potential obtained under its normal operating conditions. Thermodynamic parameters associated with electron transfer were determined for both rubredoxins (for C. pasteurianum, DeltaV degrees = -27 mL/mol, DeltaS degrees = -36 cal K-1 mol-1, and DeltaH degrees = -10 kcal/mol, and for P. furiosus, DeltaV degrees = -31 mL/mol, DeltaS degrees = -41 cal K-1 mol-1, and DeltaH degrees = -13 kcal/mol) from its pressure- and temperature-reduction potential profiles. The thermodynamic parameters for electron transfer (DeltaV degrees, DeltaS degrees, and DeltaH degrees ) for both proteins were very similar, which is not surprising considering their structural similarities and sequence homology. Despite the fact that these two proteins exhibit dramatic differences in thermostability, it appears that structural changes that confer dramatic differences in thermostability do not significantly alter electron transfer reactivity. The experimental changes in reduction potential as a function of pressure and temperature were simulated using a continuum dielectric electrostatic model (DELPHI). A reasonable estimate of the protein dielectric constant (epsilonprotein) of 6 for both rubredoxins was determined from these simulations. A discussion is presented regarding the analysis of electrostatic interaction energies of biomolecules through pressure- and temperature-controlled electrochemical studies. PMID- 10387029 TI - Guanidine hydrochloride exerts dual effects on the tryptophan synthase alpha 2 beta 2 complex as a cation activator and as a modulator of the active site conformation. AB - To characterize the conformational transitions that regulate the activity and specificity of the tryptophan synthase alpha 2 beta 2 complex, we have determined some effects of low concentrations of guanidine hydrochloride (GuHCl) and of urea on functional properties. We report the novel finding that GuHCl at low concentrations (0. 02-0.08 M) is a cation activator of the tryptophan synthase alpha 2 beta 2 complex. Molecular modeling studies show that GuH+ could bind at a previously identified cation binding site in the tryptophan synthase beta subunit. Addition of increasing concentrations of GuHCl has strikingly different effects on the rates of different reactions with L-serine or beta-chloro-L alanine in the presence or absence of indole. Spectroscopic studies demonstrate that GuHCl alters the equilibrium distribution of pyridoxal 5'-phosphate intermediates formed in reactions at the active site of the beta subunit. Data analysis shows that GuHCl binds preferentially with the conformer of the enzyme that predominates when the aldimine of L-serine is formed and shifts the equilibrium in favor of this conformer. These results provide evidence that GuHCl exerts dual effects on tryptophan synthase as a cation, stimulating activity, and as a chaotropic agent, altering the distribution of conformational states that exhibit different reaction specificities. Our finding that the nonionic urea stabilizes the aldimine of L-serine in the presence, but not in the absence, of NaCl shows that cation binding plays an important role in the conformational transitions that regulate activity and the transmission of allosteric signals between the alpha and beta sites. PMID- 10387030 TI - The amidotransferase family of enzymes: molecular machines for the production and delivery of ammonia. AB - The amidotransferase family of enzymes utilizes the ammonia derived from the hydrolysis of glutamine for a subsequent chemical reaction catalyzed by the same enzyme. The ammonia intermediate does not dissociate into solution during the chemical transformations. A well-characterized example of the structure and mechanism displayed by this class of enzymes is provided by carbamoyl phosphate synthetase (CPS). Carbamoyl phosphate synthetase is isolated from Escherichia coli as a heterodimeric protein. The smaller of the two subunits catalyzes the hydrolysis of glutamine to glutamate and ammonia. The larger subunit catalyzes the formation of carbamoyl phosphate using 2 mol of ATP, bicarbonate, and ammonia. Kinetic investigations have led to a proposed chemical mechanism for this enzyme that requires carboxy phosphate, ammonia, and carbamate as kinetically competent reaction intermediates. The three-dimensional X-ray crystal structure of CPS has localized the positions of three active sites. The nucleotide binding site within the N-terminal half of the large subunit is required for the phosphorylation of bicarbonate and subsequent formation of carbamate. The nucleotide binding site within the C-terminal domain of the large subunit catalyzes the phosphorylation of carbamate to the final product, carbamoyl phosphate. The three active sites within the heterodimeric protein are separated from one another by about 45 A. The ammonia produced within the active site of the small subunit is the substrate for reaction with the carboxy phosphate intermediate that is formed in the active site found within the N terminal half of the large subunit of CPS. Since the ammonia does not dissociate from the protein prior to its reaction with carboxy phosphate, this intermediate must therefore diffuse through a molecular tunnel that connects these two sites with one another. Similarly, the carbamate intermediate, initially formed at the active site within the N-terminal half of the large subunit, is the substrate for phosphorylation by the ATP bound to the active site located in the C-terminal half of the large subunit. A molecular passageway has been identified by crystallographic methods that apparently facilitates diffusion between these two active sites within the large subunit of CPS. Synchronization of the chemical transformations is controlled by structural perturbations among the three active sites. Molecular tunnels between distant active sites have also been identified in tryptophan synthase and glutamine phosphoribosyl pyrophosphate amidotransferase and are likely architectural features in an expanding list of enzymes. PMID- 10387031 TI - Thermodynamics of the alkaline transition of cytochrome c. AB - The apparent equilibrium constant (Kapp) of the alkaline transition (AT) of beef heart cytochrome c, obtained from pH titrations of the current intensities in cyclic voltammetry experiments, has been measured as a function of the temperature from 5 to 65 degrees C, at different ionic strength (I = 0.01-0.2 M). The temperature profile of the pKapp values is biphasic and yields two distinct sets of DeltaH degrees 'AT and DeltaS degrees 'AT values below and above approximately 40 degrees C. In the low-temperature range, the process is endothermic and is accompanied by a small positive entropy change, while at higher temperatures it becomes less endothermic and involves a pronounced entropy loss. The temperature dependence of the transition thermodynamics is most likely the result of the thermal transition of native ferricytochrome c from a low-T to an high-T conformer which occurs at alkaline pH values at a temperature comparable with above (Ikeshoji, T., Taniguchi, I., and Hawkridge, F. M. (1989) J. Electroanal. Chem. 270, 297-308; Battistuzzi, G., Borsari, M., Sola, M., and Francia, F. (1997) Biochemistry 36, 16247-16258). Thus, it is apparent that the transitions of the two native conformers to the corresponding alkaline form(s) are thermodynamically distinct processes. It is suggested that this difference arises from either peculiar transition-induced changes in the hydration sphere of the protein or to the preferential binding of different lysines to the heme iron in the two temperature ranges. Extrapolation of the Kapp values at null ionic strength allowed the determination of the thermodynamic equilibrium constants (Ka) at each temperature, hence of the "true" standard thermodynamic parameters of the transition. The pKa value at 25 degrees C was found to be 8.0. A pKapp value of 14.4 was calculated for the alkaline transition of ferrocytochrome c at 25 degrees C and I = 0.1 M. The much greater relative stabilization of the native state in the reduced as compared to the oxidized form turns out to be almost entirely enthalpic in origin, and is most likely due to the greater affinity of the methionine sulfur for the Fe(II) ion. Finally, it is found that the Debye Huckel theory fits the ionic strength dependence of the pKapp values, at least qualitatively, as observed previously for the ionic strength dependence of the reduction potential of this protein class. It is apparent that the increase in the pKapp values with increasing ionic strength is for the most part an entropic effect. PMID- 10387032 TI - Substitution of the sixth axial ligand of Rhodobacter capsulatus cytochrome c1 heme yields novel cytochrome c1 variants with unusual properties. AB - The cytochrome (cyt) c1 heme of the ubihydroquinone:cytochrome c oxidoreductase (bc1 complex) is covalently attached to two cysteine residues of the cyt c1 polypeptide chain via two thioether bonds, and the fifth and sixth axial ligands of its iron atom are histidine (H) and methionine (M), respectively. The latter residue is M183 in Rhodobacter capsulatus cyt c1, and previous mutagenesis studies revealed its critical role for the physicochemical properties of cyt c1 [Gray, K. A., Davidson, E., and Daldal, F. (1992) Biochemistry 31, 11864-11873]. In the homologous chloroplast b6f complex, the sixth axial ligand is provided by the amino group of the amino terminal tyrosine residue. To further pursue our investigation on the role played by the sixth axial ligand in heme-protein interactions, novel cyt c1 variants with histidine-lysine (K) and histidine histidine axial coordination were sought. Using a R. capsulatus genetic system, the cyt c1 mutants M183K and M183H were constructed by site-directed mutagenesis, and chromatophore membranes as well as purified bc1 complexes obtained from these mutants were characterized in detail. The studies revealed that these mutants incorporated the heme group into the mature cyt c1 polypeptides, but yielded nonfunctional bc1 complexes with unusual spectroscopic and thermodynamic properties, including shifted optical absorption maxima (lambdamax) and decreased redox midpoint potential values (Em7). The availability and future detailed studies of these stable cyt c1 mutants should contribute to our understanding of how different factors influence the physicochemical and folding properties of membrane-bound c-type cytochromes in general. PMID- 10387033 TI - Structural features of the C-terminal domain of bovine rhodopsin: a site-directed spin-labeling study. AB - Eleven single-cysteine substitution mutants have been prepared in the sequence 325-340 of rhodopsin, corresponding to the C-terminal domain. Each of the cysteine mutants was modified with a selective nitroxide reagent to introduce a spin-labeled side chain. The electron paramagnetic resonance spectra of the labeled proteins were analyzed in terms of side chain dynamics. At all sites, the spectra reflected the presence of two populations of different mobility, although one was always dominant. The mobility of the dominant population increased in a regular fashion from the palmitoylation sites at 322C and 323C to the C-terminus, where the spectra resembled those of an unfolded protein. This apparent mobility gradient is only slightly affected in mutants lacking the palmitoyl groups, suggesting that they are not responsible for physically anchoring the C-terminal peptide at one end. Binding of a monoclonal antibody to its epitope at the C terminus dramatically reduces the mobility of nearby residues, creating a local mobility gradient opposite that in the absence of the antibody. These results indicate that the C-terminal domain of rhodopsin, beyond the palmitoylation sites, is highly disordered and dynamic, resembling an unfolded peptide tethered at one end. PMID- 10387034 TI - Single-cysteine substitution mutants at amino acid positions 306-321 in rhodopsin, the sequence between the cytoplasmic end of helix VII and the palmitoylation sites: sulfhydryl reactivity and transducin activation reveal a tertiary structure. AB - As sensors for structure at the cytoplasmic face of rhodopsin, single-cysteine substitution mutants have been previously studied in the regions connecting helices III and IV and helices V and VI. In this paper we report on single cysteine substitution mutants at amino acid positions 306-321, comprising the cytoplasmic sequence between helix VII and the palmitoylation sites in rhodopsin. The cysteine opsin mutants were expressed in COS-1 cells and on treatment with 11 cis-retinal all formed the characteristic rhodopsin chromophore. Cysteines at positions 306-316 and 319 reacted in the dark with the thiol-specific reagent 4, 4'-dithiodipyridine (4-PDS) but showed a wide variation in reactivity. Cysteines at positions 317, 318, 320, and 321 showed no reaction with 4-PDS. The mutants on illumination also showed wide variations in activating GT. The mutant Y306C showed almost no GT activation, I307C and N310C were poor, and the activity of the mutants M309C, F313C, and M317C was also reduced relative to WT. The results suggest that the region comprising amino acids 306-321 is a part of a tertiary structure and that specific amino acids in this region on light-activation participate in the interaction with GT. PMID- 10387035 TI - Structural features and light-dependent changes in the sequence 306-322 extending from helix VII to the palmitoylation sites in rhodopsin: a site-directed spin labeling study. AB - Sixteen single-cysteine substitution mutants of rhodopsin were prepared in the sequence 306-321 which begins in transmembrane helix VII and ends at the palmitoylation sites at 322C and 323C. The substituted cysteine residues were modified with a selective reagent to generate a nitroxide side chain, and the electron paramagnetic resonance spectrum of each spin-labeled mutant was analyzed in terms of residue accessibility and mobility. The periodic behavior of these parameters along the sequence indicated that residues 306-314 were in a regular alpha-helical conformation representing the end of helix VII. This helix apparently extends about 1.5 turns above the surface of the membrane, with one face in strong tertiary interaction with the core of the protein. For the segment 315-321, substituted cysteine residues at 317, 318, 320, and 321 had low reactivity with the spin-label reagent. This segment has the most extensive tertiary interactions yet observed in the rhodopsin extra-membrane sequences at the cytoplasmic surface. Previous studies showed the spontaneous formation of a disulfide bond between cysteine residues at 65 and 316. This result indicates that at least some of the tertiary contacts made in the 315-321 segment are with the sequence connecting transmembrane helices I and II. Photoactivation of rhodopsin produces changes in structure detected by spin labels at 306, 313, and 316. The changes at 313 can be accounted for by movements in the adjacent helix VI. PMID- 10387036 TI - Single-cysteine substitution mutants at amino acid positions 55-75, the sequence connecting the cytoplasmic ends of helices I and II in rhodopsin: reactivity of the sulfhydryl groups and their derivatives identifies a tertiary structure that changes upon light-activation. AB - Cysteines were introduced, one at a time, at amino acid positions 55-75 in the cytoplasmic region connecting helices I and II in rhodopsin. In each of the 21 cysteine mutants, the reactive native cysteine residues (C140 and C316) were replaced by serine. Except for N55C, all mutants formed rhodopsin-like chromophores and had normal photobleaching characteristics. The efficiency of GT activation was reduced only for K66C, K67C, L68C, and P71C. The reactivity of the substituted cysteine in each mutant toward 4, 4'-dithiodipyridine (4-PDS) was investigated in the dark. The mutants F56C to L59C and I75C were unreactive to 4 PDS under the conditions used, suggesting that they are embedded in the micelle or protein interior. The mutants V63C, H65C-T70C, and N73C reacted rapidly, while the remainder of the mutants reacted more slowly, and varied in reactivity with sequence position. For the mutants derivatized with 4-PDS, the rate of release of thiopyridone from the resulting thiopyridinyl-cysteine disulfide bond by dithiothreitol was investigated in the dark and in the light. Marked changes in the rates of thiopyridone release in the light were found at specific sites. Collectively, the data reveal tertiary interactions of the residues in the sequence investigated and demonstrate structural changes due to photoactivation. PMID- 10387037 TI - Structural features and light-dependent changes in the sequence 59-75 connecting helices I and II in rhodopsin: a site-directed spin-labeling study. AB - Twenty-one single-cysteine substitution mutants were prepared in the sequence 56 75 between transmembrane helices I and II at the cytoplasmic surface of bovine rhodopsin. Each mutant was reacted with a sulfhydryl-specific reagent to produce a nitroxide side chain. The electron paramagnetic resonance of the labeled proteins in dodecyl maltoside solution was analyzed to provide the relative mobility and accessibility of the nitroxide side chain to both polar and nonpolar paramagnetic reagents. The results indicate that the hydrophobic-water interface of the micelle intersects helices I and II near residues 64 and 71, respectively. Thus, the sequence 64-71 is in the aqueous phase, while 56-63 and 72-75 lie in the transmembrane helices I and II, respectively. The lipid-facing surfaces on transmembrane helices I and II near the cytoplasmic surface correspond to approximately 180 degrees and 90 degrees of arc on the helical surfaces, respectively. Photoactivation of rhodopsin produced changes in structure in the region investigated, primarily around helix II. However, these changes are much smaller than those noted by spin labels in helix VI (Altenbach, C., Yang, K., Farrens, D., Farahbakhsh, Z., Khorana, H. G., and Hubbell, W. L. (1996) Biochemistry 35, 12470). PMID- 10387039 TI - Electrostatic and aromatic microdomains within the binding-site crevice of the D2 receptor: contributions of the second membrane-spanning segment. AB - The binding-site of the dopamine D2 receptor, like that of other homologous G protein-coupled receptors, is contained within a water-accessible crevice formed among its seven membrane-spanning segments. Using the substituted cysteine accessibility method (SCAM), we previously mapped the residues in the third, fifth, sixth, and seventh membrane-spanning segments that contribute to the surface of this binding-site crevice. We have now mutated to cysteine, one at a time, 22 consecutive residues in the second membrane-spanning segment (M2) and expressed the mutant receptors in HEK 293 cells. Eleven of these mutants reacted with charged, hydrophilic, lipophobic, sulfhydryl-specific reagents, added extracellularly, and 9 of these 11 were protected from reaction by a reversible dopamine antagonist, sulpiride. We infer that the side chains of the residues at the 11 reactive loci (D80, L81, V83, V87, P89, W90, V91, V92, L94, E95, V96) are on the water-accessible surface of the binding-site crevice and that 9 of these are occluded by bound antagonist. The pattern of accessibility suggests an alpha helical conformation. A broadening of the angle of accessibility near the binding site is consistent with the presence of a kink at Pro89. On the basis of the enhanced rates of reaction of positively charged sulfhydryl reagents, we infer the presence of an electrostatic microdomain composed of three acidic residues in M2 and the adjacent M3 that could attract and orient cationic ligands. Furthermore, based on the enhanced reactivity of the hydrophobic cation containing reagent, we infer the presence of an aromatic microdomain formed between M2, M3, and M7. PMID- 10387038 TI - Molecular determinants of the reversible membrane anchorage of the G-protein transducin. AB - Transducin is a heterotrimer formed by a fatty acylated alpha-subunit and a farnesylated betagamma-subunit. The role of these two covalent modifications and of adjacent hydrophobic and charged amino acid residues in reversible anchoring at disk model membranes is investigated at different pH values, salt concentrations, and lipid packing densities using the monolayer expansion technique and CD spectroscopy. The heterotrimer only binds if the acetylated alpha-subunit is transformed into its surface-active form by divalent cations. In the presence of salts the alpha(GDP)-subunit, the betagamma-complex, and the heterotrimer bind to POPC monolayers at 30 mN/m, estimated to mimic the lateral packing density of disk membranes, with apparent binding constants of Kapp = (1.1 +/- 0.3) x 10(6) M-1 (reflecting the penetration of the fatty acyl chain together with approximately three adjacent hydrophobic amino acid residues), Kapp = (3.5 +/- 0.5) x 10(6) M-1 (reflecting the penetration of the farnesyl chain), and Kapp = (1.6 +/- 0.3) x 10(6) M-1 (reflecting a major contribution of the alpha(GDP) subunit with only a minor contribution from the betagamma-complex). The apparent binding constant of the alpha(GTP)-subunit is distinctly smaller than that of the alpha(GDP)-subunit. Binding to negatively charged POPC/POPG (75/25 mole/mole) monolayers is reinforced by 2-3 cationic residues for the betagamma-complex. The alpha-subunit shows no electrostatic attraction and the heterotrimer shows even a slight electrostatic repulsion which becomes the dominating force in the absence of salts. PMID- 10387040 TI - Crystal structures of thrombin complexed to a novel series of synthetic inhibitors containing a 5,5-trans-lactone template. AB - The binding modes of four active site-directed, acylating inhibitors of human alpha-thrombin have been determined using X-ray crystallography. These inhibitors (GR157368, GR166081, GR167088, and GR179849) are representatives of a series utilizing a novel 5, 5-trans-lactone template to specifically acylate Ser195 of thrombin, resulting in an acyl complex. In each case the crystal structure of the complex reveals a binding mode which is consistent with the formation of a covalent bond between the ring-opened lactone of the inhibitor and residue Ser195. Improvements in potency and selectivity of these inhibitors for thrombin are rationalized on the basis of the observed protein/inhibitor interactions identified in these complexes. Occupation of the thrombin S2 and S3 pockets is shown to be directly correlated with improved binding and a degree of selectivity. The binding mode of GR179849 to thrombin is compared with the thrombin/PPACK complex [Bode, W., Turk, D., and Karshikov, A. (1992) Protein Sci. 1, 426-471] as this represents the archetypal binding mode for a thrombin inhibitor. This series of crystal structures is the first to be reported of synthetic, nonpeptidic acylating inhibitors bound to thrombin and provides details of the molecular recognition features that resulted in nanomolar potency. PMID- 10387041 TI - Molecular recognition of macrocyclic peptidomimetic inhibitors by HIV-1 protease. AB - High-resolution crystal structures are described for seven macrocycles complexed with HIV-1 protease (HIVPR). The macrocycles possess two amides and an aromatic group within 15-17 membered rings designed to replace N- or C-terminal tripeptides from peptidic inhibitors of HIVPR. Appended to each macrocycle is a transition state isostere and either an acyclic peptide, nonpeptide, or another macrocycle. These cyclic analogues are potent inhibitors of HIVPR, and the crystal structures show them to be structural mimics of acyclic peptides, binding in the active site of HIVPR via the same interactions. Each macrocycle is restrained to adopt a beta-strand conformation which is preorganized for protease binding. An unusual feature of the binding of C-terminal macrocyclic inhibitors is the interaction between a positively charged secondary amine and a catalytic aspartate of HIVPR. A bicyclic inhibitor binds similarly through its secondary amine that lies between its component N-terminal and C-terminal macrocycles. In contrast, the corresponding tertiary amine of the N-terminal macrocycles does not interact with the catalytic aspartates. The amine-aspartate interaction induces a 1.5 A N-terminal translation of the inhibitors in the active site and is accompanied by weakened interactions with a water molecule that bridges the ligand to the enzyme, as well as static disorder in enzyme flap residues. This flexibility may facilitate peptide cleavage and product dissociation during catalysis. Proteases [Aba67,95]HIVPR and [Lys7,Ile33,Aba67,95]HIVPR used in this work were shown to have very similar crystal structures. PMID- 10387042 TI - Mechanistic insights into the inhibition of serine proteases by monocyclic lactams. AB - Although originally discovered as inhibitors of pencillin-binding proteins, beta lactams have more recently found utility as serine protease inhibitors. Indeed through their ability to react irreversibly with nucleophilic serine residues they have proved extraordinarily successful as enzyme inhibitors. Consequently there has been much speculation as to the reason for the general effectiveness of beta-lactams as antibacterials or inhibitors of hydrolytic enzymes. The interaction of analogous beta- and gamma-lactams with a serine protease was investigated. Three series of gamma-lactams based upon monocyclic beta-lactam inhibitors of elastase [Firestone, R. A. et al. (1990) Tetrahedron 46, 2255 2262.] but with an extra methylene group inserted between three of the bonds in the ring were synthesized. Their interaction with porcine pancreatic elastase and their efficacy as inhibitors were evaluated through the use of kinetic, NMR, mass spectrometric, and X-ray crystallographic analyses. The first series, with the methylene group inserted between C-3 and C-4 of the beta-lactam template, were readily hydrolyzed but were inactive or very weakly active as inhibitors. The second series, with the methylene group between C-4 and the nitrogen of the beta lactam template, were inhibitory and reacted reversibly with PPE to form acyl enzyme complexes, which were stable with respect to hydrolysis. The third series, with the methylene group inserted between C-2 and C-3, were not hydrolyzed and were not inhibitors consistent with lack of binding to PPE. Comparison of the crystal structure of the acyl-enzyme complex formed between PPE and a second series gamma-lactam and that formed between PPE and a peptide [Wilmouth, R. C., et al. (1997) Nat. Struct. Biol. 4, 456-462.] reveals why the complexes formed with this series were resistant to hydrolysis and suggests ways in which stable acyl-enzyme complexes might be obtained from monocyclic gamma-lactam-based inhibitors. PMID- 10387043 TI - Crystal structure of substrate complexes of methylmalonyl-CoA mutase. AB - X-ray crystal structures of methylmalonyl-CoA mutase in complexes with substrate methylmalonyl-CoA and inhibitors 2-carboxypropyl-CoA and 3-carboxypropyl-CoA (substrate and product analogues) show that the enzyme-substrate interactions change little during the course of the rearrangement reaction, in contrast to the large conformational change on substrate binding. The substrate complex shows a 5'-deoxyadenine molecule in the active site, bound weakly and not attached to the cobalt atom of coenzyme B12, rotated and shifted from its position in the substrate-free adenosylcobalamin complex. The position of Tyralpha89 close to the substrate explains the stereochemical selectivity of the enzyme for (2R) methylmalonyl-CoA. PMID- 10387045 TI - Low-temperature stabilization and spectroscopic characterization of the dioxygen complex of the ferrous neuronal nitric oxide synthase oxygenase domain. AB - Nitric oxide (NO), an intercellular messenger and an immuno-cytotoxic agent, is synthesized by the family of nitric oxide synthases (NOS), which are thiolate ligated, heme-containing monooxygenases that convert L-Arg to L-citrulline and NO in a tetrahydrobiopterin (BH4)-dependent manner, using NADPH as the electron donor. The dioxygen complex of the ferrous enzyme has been proposed to be a key intermediate in the NOS catalytic cycle. In this study, we have generated a stable ferrous-O2 complex of the oxygenase domain of rat neuronal NOS (nNOS) by bubbling O2 through a solution of the dithionite-reduced enzyme at -30 degrees C in a cryogenic solvent containing 50% ethylene glycol. The most stable dioxygen complex is obtained using the oxygenase domain which has been preincubated for an extended length of time at 4 degrees C with BH4/dithiothreitol and NG-methyl-L arginine, a substrate analogue inhibitor. The O2 complex of the nNOS oxygenase domain thus prepared exhibits UV-visible absorption (maxima at 419 and 553 nm, shoulder at approximately 585 nm) and magnetic circular dichroism spectra that are nearly identical to those of ferrous-O2 cytochrome P450-CAM. Our spectral data are noticeably blue-shifted from those seen at 10 degrees C for a short lived transient species (lambdamax = 427 nm) for the nNOS oxygenase domain using stopped-flow rapid-scanning spectroscopy [Abu-Soud, H. M., Gachhui, R., Raushel, F. M., and Stuehr, D. J. (1997) J. Biol. Chem. 272, 17349], but somewhat similar to those of a relatively stable O2 adduct of L-Arg-free full-length nNOS (lambdamax = 415-416.5 nm) generated at -30 degrees C [Bec, N., Gorren, A. C. F., Voelder, C., Mayer, B., and Lange, R. (1998) J. Biol. Chem. 273, 13502]. Compared with ferrous-O2 P450-CAM, however, the ferrous-O2 adduct of the nNOS oxygenase domain is considerably more autoxidizable and the O2-CO exchange reaction is noticeably slower. The generation of a stable ferrous-O2 adduct of the nNOS oxygenase domain, as described herein, will facilitate further mechanistic and spectroscopic investigations of this important intermediate. PMID- 10387044 TI - Binding of biliverdin, bilirubin, and thyroid hormones to lipocalin-type prostaglandin D synthase. AB - Lipocalin-type prostaglandin D synthase is a major protein of the cerebrospinal fluid and was originally known as beta-trace. We investigated the binding ability of prostaglandin D synthase toward bile pigments, thyroid hormones, steroid hormones, and fatty acids in this present study. We found that the recombinant enzyme binds bile pigments and thyroid hormones, resulting in quenching of the intrinsic tryptophan fluorescence, the appearance of induced circular dichroism of the lipophilic ligands, and a red shift of the absorption spectra of bilirubin and biliverdin. The binding of prostaglandin D synthase to lipophilic ligands was also demonstrated by the resonant mirror technique and surface plasmon resonance detection. The dissociation constants were calculated to be 33 nM, 37 nM, 660 nM, 820 nM, and 2.08 microM for biliverdin, bilirubin, L-thyroxine, 3,3',5'-triiodo-L thyronine, and 3,3', 5-triiodo-L-thyronine, respectively. Biliverdin and bilirubin underwent a shift in their absorption peaks from 375 to 380 nm and from 439 to 446 nm, respectively, after binding to prostaglandin D synthase. Bilirubin bound to the enzyme showed a bisignate CD spectrum with a (-) Cotton effect at 422 nm and a (+) Cotton effect at 472 nm, indicating a right-handed chirality. The ligands also inhibited prostaglandin D synthase activity noncompetitively in a concentration-dependent manner, with IC50 values between 3.9 and 10. 9 microM. Epididymal retinoic acid-binding protein and beta-lactoglobulin, two other lipocalin proteins that bind retinoids such as prostaglandin D synthase, did not show any significant interaction with bile pigments or thyroid hormones. These results show that prostaglandin D synthase binds small lipophilic ligands with a specificity distinct from that of other lipocalins. PMID- 10387046 TI - Solid state NMR studies of hydrogen bonding in a citrate synthase inhibitor complex. AB - The ionization state and hydrogen bonding environment of the transition state analogue (TSA) inhibitor, carboxymethyldethia coenzyme A (CMX), bound to citrate synthase have been investigated using solid state NMR. This enzyme-inhibitor complex has been studied in connection with the postulated contribution of short hydrogen bonds to binding energies and enzyme catalysis: the X-ray crystal structure of this complex revealed an unusually short hydrogen bond between the carboxylate group of the inhibitor and an aspartic acid side chain [Usher et al. (1994) Biochemistry 33, 7753-7759]. To further investigate the nature of this short hydrogen bond, low spinning speed 13C NMR spectra of the CMX-citrate synthase complex were obtained under a variety of sample conditions. Tensor values describing the chemical shift anisotropy of the carboxyl groups of the inhibitor were obtained by simulating MAS spectra (233 +/- 4, 206 +/- 5, and 105 +/- 2 ppm vs TMS). Comparison of these values with our previously reported database and ab initio calculations of carbon shift tensor values clearly indicates that the carboxyl is deprotonated. New data from model compounds suggest that hydrogen bonds in a syn arrangement with respect to the carboxylate group have a pronounced effect upon the shift tensors for the carboxylate, while anti hydrogen bonds, regardless of their length, apparently do not perturb the shift tensors of the carboxyl group. Thus the tensor values for the enzyme inhibitor complex could be consistent with either a very long syn hydrogen bond or an anti hydrogen bond; the latter would agree very well with previous crystallographic results. Two-dimensional 1H-13C heteronuclear correlation spectra of the enzyme-inhibitor complex were obtained. Strong cross-peaks were observed from the carboxyl carbon to proton(s) with chemical shift(s) of 22 +/- 5 ppm. Both the proton chemical shift and the intensity of the cross-peak indicate a very short hydrogen bond to the carboxyl group of the inhibitor, the C.H distance based upon the cross-peak intensity being 2.0 +/- 0.4 A. This proton resonance is assigned to Hdelta2 of Asp 375, on the basis of comparison with crystal structures and the fact that this cross-peak was absent in the heteronuclear correlation spectrum of the inhibitor-D375G mutant enzyme complex. In summary, our NMR studies support the suggestion that a very short hydrogen bond is formed between the TSA and the Asp carboxylate. PMID- 10387047 TI - A novel, definitive test for substrate channeling illustrated with the aspartate aminotransferase/malate dehydrogenase system. AB - A novel method is presented that establishes definitively the existence or nonexistence of direct metabolite transfer between consecutive enzymes in a metabolic sequence. The procedure is developed with the specific example of channeling of oxaloacetate between Escherichia coli aspartate aminotransferase (AATase) and malate dehydrogenase (MDH). The assay is carried out in the presence of a large excess of inactive variants of AATase. These mutants would outcompete the much smaller quantities of wild-type AATase for any docking sites on MDH and thus decrease the rate of the coupled L-aspartate to oxaloacetate to malate sequence only if the direct metabolite transfer mechanism is operative. The results show that oxaloacetate is not transferred directly from AATase to MDH because no decrease in rate was observed in the presence of approximately 100 microM inactive mutants. This concentration is 10 times the physiological AATase concentration, which was determined in this work. The methodology can be applied generally. PMID- 10387048 TI - Direct measurement of the pKa of aspartic acid 26 in Lactobacillus casei dihydrofolate reductase: implications for the catalytic mechanism. AB - The ionization state of aspartate 26 in Lactobacillus casei dihydrofolate reductase has been investigated by selectively labeling the enzyme with [13Cgamma] aspartic acid and measuring the 13C chemical shifts in the apo, folate enzyme, and dihydrofolate-enzyme complexes. Our results indicate that no aspartate residue has a pKa greater than approximately 4.8 in any of the three complexes studied. The resonance of aspartate 26 in the dihydrofolate-enzyme complex has been assigned by site-directed mutagenesis; aspartate 26 is found to have a pKa value of less than 4 in this complex. Such a low pKa value makes it most unlikely that the ionization of this residue is responsible for the observed pH profile of hydride ion transfer [apparent pKa = 6.0; Andrews, J., Fierke, C. A., Birdsall, B., Ostler, G., Feeney, J., Roberts, G. C. K., and Benkovic, S. J. (1989) Biochemistry 28, 5743-5750]. Furthermore, the downfield chemical shift of the Asp 26 (13)Cgamma resonance in the dihydrofolate-enzyme complex provides experimental evidence that the pteridine ring of dihydrofolate is polarized when bound to the enzyme. We propose that this polarization of dihydrofolate acts as the driving force for protonation of the electron-rich O4 atom which occurs in the presence of NADPH. After this protonation of the substrate, a network of hydrogen bonds between O4, N5 and a bound water molecule facilitates transfer of the proton to N5 and transfer of a hydride ion from NADPH to the C6 atom to complete the reduction process. PMID- 10387049 TI - New strategy for the conformational analysis of carbohydrates based on NOE and 13C NMR coupling constants. Application to the flexible polysaccharide of Streptococcus mitis J22. AB - For complex oligosaccharides, which are relatively rigid with modest excursions from a single minimum energy conformation, it is straightforward to build conformational models from NOE data. Other oligosaccharides are more flexible with transitions between distinct minima separated by substantial energy barriers. We show that modeling based on scalar coupling data is superior to NOE based modeling for the latter case. Long range 13C-13C and 13C-1H coupling constants measured for the heptasaccharide repeating subunit of the cell wall polysaccharide from Streptococcus mitis J22 are correlated with individual glycosidic dihedral angles, effectively uncoupling the degrees of freedom of the oligosaccharide and allowing a search for combinations of dihedral angles which are energetically reasonable, i.e., with no bad van der Waals contacts, and which can be combined to satisfy all the measured J values. Allowed values of the individual angles can then be combined to search for overall oligosaccharide conformations which contribute to the ensemble. We show that while the polysaccharide from S. mitis J22 is flexible, requiring multiple conformations, most of the flexibility is localized to a few bonds and only a rather small number of conformations is required to reproduce the experimental NOE and scalar coupling data. PMID- 10387050 TI - Quantitative analysis of phospholipids in functionally important membrane domains from RBL-2H3 mast cells using tandem high-resolution mass spectrometry. AB - We recently showed that ligand-mediated cross-linking of FcepsilonRI, the high affinity receptor for immunoglobulin E, on RBL-2H3 mast cells results in its co isolation with detergent-resistant membranes (DRM) and its consequent tyrosine phosphorylation by the co-localized tyrosine kinase Lyn that is a critical early event in signaling by this receptor [Field et al. (1997) J. Biol. Chem. 272, 4276 4280]. As part of efforts to determine the structural bases for these interactions, we examined the phospholipid composition of DRM vesicles isolated from RBL-2H3 cells under conditions that preserve FcepsilonRI association. We used positive and negative mode electrospray Fourier transform ion cyclotron resonance mass spectrometry to compare quantitatively the phospholipid composition of isolated DRM to that of total cell lipids and to a plasma membrane preparation. From these analyses, over 90 different phospholipid species were spectrally resolved and unambiguously identified; more than two-thirds of these were determined with a precision of +/-0.5% (absolute) or less. Quantitative characterization of lipid profiles shows that isolated DRM are substantially enriched in sphingomyelin and in glycerophospholipids with a higher degree of saturation as compared to total cellular lipids. Plasma membrane vesicles isolated from RBL-2H3 cells by chemically induced blebbing exhibit a degree of phospholipid saturation that is intermediate between DRM and total cellular lipids, and significant differences in the headgroup distribution between DRM and plasma membranes vesicles are observed. DRM from cells with cross-linked FcepsilonRI exhibit a larger ratio of polyunsaturated to saturated and monounsaturated phospholipids than those from unstimulated cells. Our results support and strengthen results from previous studies suggesting that DRM have a lipid composition that promotes liquid-ordered structure. Furthermore, they demonstrate the potential of mass spectrometry for examining the role of membrane structure in receptor signaling and other cellular processes. PMID- 10387051 TI - Amino acid sequences within the alpha subunit of integrin alpha M beta 2 (Mac-1) critical for specific recognition of C3bi. AB - Phagocytosis of opsonized particles by neutrophils and monocytes plays a central role in host defense mechanisms against foreign pathogens. This process depends on the interaction between C3bi, a degradation product derived from activation of the complement system, and the alpha M beta 2 (CD11b/CD18, Mac-1) receptor, the major integrin on neutrophils. Previous studies had established a central role for the I domain, a stretch of approximately 200 amino acids within the alpha M subunit in the binding of C3bi, as well as many other alpha M beta 2 ligands. The present study was undertaken to establish the molecular basis of C3bi recognition by alpha M beta 2. The strategy employed the use of a series of mutant receptors in which short segments of the I domain of alpha M were switched to the corresponding segments of alpha L, which is structurally very similar but does not bind C3bi. We report three major findings: (1) The C3bi binding pocket is composed of three regions, P147-R152, P201-K217, and K245-R261 of alpha M, which surround the cation binding site within the MIDAS motif of the I domain. (2) Within the latter segment, K245 plays a critical role in mediating C3bi binding to alpha M beta 2. Mutation of K245 to Ala significantly reduced C3bi binding but had no effect on binding of another alpha M beta 2 I domain ligand, NIF. (3) Blocking of C3bi binding to alpha M beta 2 by monoclonal antibodies is achieved through two different mechanisms: direct competition for the ligand binding site or induction of conformational changes. Overall, these studies support the hypothesis that many of the ligands of alpha M beta 2 bind to overlapping but not identical sites within the I domain. Although the same short structural segments within the I domain may be involved in binding, different amino acids within these segments may contact different ligands. PMID- 10387052 TI - Interaction of the vaccinia virus nucleoside triphosphate phosphohydrolase I with linear oligonucleotides. AB - Vaccinia virus nucleoside triphosphate phosphohydrolase I (NPH I) serves as the ATPase activity employed in early gene transcription termination [Deng, L., and Shuman, S. (1998) Genes Dev. 12, 538-546; Christen, L. M., et al. (1998) Virology 245, 360-371]. Since ATPase activity requires binding of single-stranded DNA, a full understanding of the mechanism of oligonucleotide activation is essential for the elucidation of its role in transcription termination. To initiate detailed structure-function studies of NPH I, we undertook combined kinetic and binding analyses of the interaction of linear oligonucleotides with NPH I. In the presence of single-stranded DNA, ATP exhibits complex saturation kinetics. The apparent Km for ATP is independent of DNA concentration, demonstrating that ssDNA binding alters the kcat for the reaction. Linear ssDNA oligonucleotides from 18 to 48 nucleotides in length stimulated activity in a saturatable fashion. As the oligonucleotide length increases, the Kact decreases and the Vmax increases. The increase in affinity is paralleled by an increase in the level of binding as measured by EMSA. The kinetic activation observed for 36-nucleotide ssDNA is dependent upon ATP concentration. At low ATP levels, sigmoidal saturation kinetics are observed, while at saturating ATP levels, near-hyperbolic kinetics are seen, suggesting that NPH I may adopt two conformational states. Linear oligonucleotides 18, 24, and 36 bases in length bind one, two, and three molecules of NPH I maximally, respectively, indicating that the NPH I binding site is no more than 12 bases in length. In contrast, single-stranded RNA does not stimulate ATPase activity, yet RNA binds as well as DNA of a similar length. Both RNA and DNA can be photo-cross-linked to NPH I by UV light. ssDNA and ssRNA cross-compete in UV photo-cross-linking to NPH I, indicating that both oligonucleotides share a common binding site. ssRNA prevents ssDNA activation of ATPase activity, confirming that both oligonucleotides bind to the kinetically important oligonucleotide activation site on NPH I. ssDNA inhibits transcription termination in vitro. Inhibition is overcome by adding NPH I, demonstrating that oligonucleotide inhibition is mediated through NPH I. PMID- 10387053 TI - Binding kinetics and footprinting of TaqI endonuclease: effects of metal cofactors on sequence-specific interactions. AB - Restriction endonucleases achieve sequence-specific recognition and strand cleavage through the interplay of base, phosphate backbone, and metal cofactor interactions. In this study, we investigate the binding kinetics of TaqI endonuclease using the wild-type enzyme and a binding proficient, catalysis deficient mutant TaqI-D137A both in the absence of a metal cofactor and in the presence of Mg2+ or Ca2+. As demonstrated by gel mobility shift analyses, TaqI endonuclease requires a metal cofactor for achieving high-affinity specific binding to its cognate sequence, TCGA. In the absence of a metal cofactor, the enzyme binds all DNA sequences (TaqI cognate site, star site, and nonspecific site) with essentially equal affinity, thereby exhibiting little discrimination. The dissociation constant of the cognate sequence in the presence of Mg2+ at 60 degrees C is 0. 26 nM, a value comparable to our previously reported Km of 0.5 nM measured under steady-state conditions. The TaqI-TCGA-Mg2+ complex is stable, with a half-life of 21 min at 60 degrees C. The boundary of the protein-DNA interface is approximated to be about 18 bp as determined by DNase I footprinting. Data from this study support the notion that a metal cofactor plays a critical role for achieving sequence-specific discrimination in a subset of nucleases, including TaqI, EcoRV, and others. PMID- 10387054 TI - Misacylation of tRNALys with noncognate amino acids by lysyl-tRNA synthetase. AB - Lysyl-tRNA synthetase (LysRS), a class II enzyme whose major function is to provide Lys-tRNALys for protein synthesis, also catalyzes aminoacylation of tRNALys with arginine, threonine, methionine, leucine, alanine, serine, and cysteine. The limited selectivity in the tRNA aminoacylation reaction appears to be due to inefficient editing of some amino acids (Met, Leu, Cys, Ala, Thr) by a pre-transfer mechanism or the absence of editing of other amino acids (Arg and Ser). Purified Arg-tRNALys, Thr-tRNALys, and Met-tRNALys were essentially not deacylated by LysRS, indicating that the enzyme does not possess a post-transfer editing mechanism. However, LysRS possesses an efficient pre-transfer editing mechanism which prevents misacylation of tRNALys with ornithine. A novel feature of this editing reaction is that ornithine lactam is formed by the facile cyclization of ornithyl adenylate. PMID- 10387055 TI - Steady-state kinetic characterization of RB69 DNA polymerase mutants that affect dNTP incorporation. AB - The function of six highly conserved residues (Arg482, Lys483, Lys486, Lys560, Asn564, and Tyr567) in the fingers domain of bacteriophage RB69 DNA polymerase (RB69 gp43) were analyzed by kinetic studies with mutants in which each of these residues was replaced with Ala. Our results suggest that Arg482, Lys486, Lys560, and Asn564 contact the incoming dNTP during the nucleotidyl transfer reaction as judged by variations in apparent Km and kcat values for dNTP incorporation by these mutants compared to those for the exonuclease deficient parental polymerase under steady-state conditions. On the basis of our studies, as well as on the basis of the crystal structure of RB69 gp43, we propose that a conformational change in the fingers domain, which presumably occurs prior to polymerization, brings the side chains of Arg482, Lys486, Lys560, and Asn564 into the vicinity of the primer-template terminus where they can contact the triphosphate moiety of the incoming dNTP. In particular, on the basis of structural studies reported for the "closed" forms of two other DNA polymerases and from the kinetic studies reported here, we suggest that (i) Lys560 and Asn564 contact the nonbonding oxygens of the alpha and beta phosphates, respectively, and (ii) both Arg482 and Lys486 contact the gamma phosphate oxygens of the incoming dNTP of RB69 gp43 prior to the nucleotidyl transfer reaction. We also found that Ala substitutions at each of these four RB69 gp43 sites could incorporate dGDP as a substrate, although with markedly reduced efficiency compared to that with dGTP. In contrast in the parental exo- background, the K483A and Y567A substituted enzymes could not use dGDP as a substrate for primer extension. These results, taken together, are consistent with the putative roles of the four conserved residues in RB69 gp43 as stated above. PMID- 10387056 TI - Influence of proline residues on the antibacterial and synergistic activities of alpha-helical peptides. AB - To investigate the influence of proline residues on the activity of alpha-helical peptides, variants were synthesized with insertions of proline residues to create peptides without proline, or with one or two prolines. The influence of the proline-induced bends was assessed by circular dichroism in the presence of liposomes, and the ability of the peptides to kill microorganisms, to permeabilize the outer and cytoplasmic membranes of Escherichia coli, to bind to liposomes, to form channels in planar lipid bilayers, and to synergize with conventional antibiotics. Representative peptides adopted alpha-helical conformations in phosphatidylcholine/phosphatidylglycerol (POPC/POPG, 7:3) liposomes as well as in 60% trifluoroethanol solution, as revealed by circular dichroism (CD) spectroscopy. However, the percent of helicity decreased as the number of proline residues increased. Tryptophan fluorescence spectroscopy showed that all of these peptides inserted into the membranes of liposomes as indicated by a blue shift in the emission maximum and an increase in the fluorescence intensity of the single tryptophan at residue 2. Quenching experiments further prove that the tryptophan residue was no longer accessible to the aqueous quencher KI. The peptide that lacked proline exhibited the highest activity [minimal inhibitory concentrations (MICs) of 0.5-4 microg/mL] against all tested Gram-negative and Gram-positive bacteria, but was hemolytic at 8 microg/mL. The single-proline peptides exhibited intermediate antibacterial activity. Peptides with two proline residues were even less active with moderate MICs only against E. coli. With only one exception from each group, the peptides were nonhemolytic. The ability of the peptides to demonstrate synergy in combination with conventional antibiotics increased as the antibacterial effectiveness decreased. All peptides bound to bacterial lipopolysaccharide and permeabilized the outer membrane of E. coli to similar extents. However, their ability to permeabilize the cytoplasmic membrane of E. coli as assessed by the unmasking of cytoplasmic beta-galactosidase decreased substantially as the number of proline residues increased. Correspondingly, increasing the number of proline residues caused a decreased ability to form channels in planar lipid bilayers, and the hemolytic, proline-free peptide tended to cause rapid breakage of planar membranes. Thus, the number of bends created by insertion of proline residues is an important determinant of antimicrobial, hemolytic, and synergistic activity. PMID- 10387057 TI - Unusual electrostatic effects on binding of C1q to anionic liposomes: role of anionic phospholipid domains and their line tension. AB - The binding of 125I-C1q to anionic liposomes was studied as a function of protein concentration, pH, ionic strength, and anionic lipid composition. The maximum amount of protein bound per micromole of lipid was very sensitive to electrostatic factors, increasing strongly with decreased pH and ionic strength or increased anionic lipid content. The apparent association constant was independent of these electrostatic factors, however, in marked contrast to studies on basic peptide binding to anionic lipid vesicles. Microscopic observations of large unilamellar liposomes containing fluorescently labeled C1q or phosphatidylglycerol demonstrated, under conditions causing strong electrostatic interactions, that C1q and anionic lipids colocalized into domains whose radii of curvature were higher than that of the surrounding lipid. These domains were observed to bud and pinch off into brightly fluorescent vesicles. We propose a model for all of these observations in which the line tension or edge energy at the boundary of the domain resists its increase in circumference as the domain grows by electrostatic effects on binding, eventually resulting in vesiculation. We propose that under favorable electrostatic conditions, as larger domains form the edge energy balances the increases in the electrostatic contribution to binding, resulting in a net binding energy independent of electrostatic factors. PMID- 10387058 TI - Use of free energy relationships to probe the individual steps of hydroxylation of p-hydroxybenzoate hydroxylase: studies with a series of 8-substituted flavins. AB - We report Hammett correlations, using 8-substituted flavins, to clarify the mechanism of hydroxylation by p-hydroxybenzoate hydroxylase (PHBH). The 8 position of the FAD isoalloxazine ring was chosen for modifications, because in PHBH it has minimal interactions with the protein, and it is accessible to solvent and away from the site of hydroxylation. Although two intermediates, a flavin-C4a-hydroperoxide and a flavin-C4a-hydroxide, are known to participate in hydroxylation, the mechanism of oxygen transfer remains controversial. Mechanisms as diverse as electrophilic aromatic substitution, diradical formation, and isoalloxazine ring opening have been proposed. In the studies reported here, it was possible to monitor spectrally each of the individual steps involved in hydroxylation, because the FAD cofactor acts as a reporter group. Thus, with PHBH, substituted separately with nine derivatives of FAD altered in the 8 position, quantitative structure-reactivity relationships (QSAR) have been applied to probe the mechanisms of formation of the flavin-C4a-hydroperoxide, the conversion to the flavin-C4a-hydroxide with concomitant oxygen transfer to the substrate, and the dehydration of the flavin-C4a-hydroxide to form oxidized FAD. The individual chemical steps in the mechanism of PHBH were not altered when using any of the modified flavins, and normal products were obtained; however, the rates of individual steps were affected, and depended on the electronic properties of the 8-substituent. Increased hydroxylation rates were observed when a more electrophilic flavin-C4a-hydroperoxide (i.e., with an electron-withdrawing substituent at the 8-position) is bound to PHBH. On the basis of QSAR analysis, we conclude that the mechanism of the hydroxylation step is best described by electrophilic aromatic substitution. PMID- 10387059 TI - Thermal versus guanidine-induced unfolding of ubiquitin. An analysis in terms of the contributions from charge-charge interactions to protein stability. AB - We have characterized the guanidine-induced unfolding of both yeast and bovine ubiquitin at 25 degrees C and in the acidic pH range on the basis of fluorescence and circular dichroism measurements. Unfolding Gibbs energy changes calculated by linear extrapolation from high guanidine unfolding data are found to depend very weakly on pH. A simple explanation for this result involves the two following assumptions: (1) charged atoms of ionizable groups are exposed to the solvent in native ubiquitin (as supported by accessible surface area calculations), and Gibbs energy contributions associated with charge desolvation upon folding (a source of pK shifts) are small; (2) charge-charge interactions (another source of pK shifts upon folding) are screened out in concentrated guanidinium chloride solutions. We have also characterized the thermal unfolding of both proteins using differential scanning calorimetry. Unfolding Gibbs energy changes calculated from the calorimetric data do depend strongly on pH, a result that we attribute to the pH dependence of charge-charge interactions (not eliminated in the absence of guanidine). In fact, we find good agreement between the difference between the two series of experimental unfolding Gibbs energy changes (determined from high guanidine unfolding data by linear extrapolation and from thermal denaturation data in the absence of guanidine) and the theoretical estimates of the contribution from charge-charge interactions to the Gibbs energy change for ubiquitin unfolding obtained by using the solvent-accessibility-corrected Tanford Kirkwood model, together with the Bashford-Karplus (reduced-set-of-sites) approximation. This contribution is found to be stabilizing at neutral pH, because most charged groups on the native protein interact mainly with groups of the opposite charge, a fact that, together with the absence of large charge desolvation contributions, may explain the high stability of ubiquitin at neutral pH. In general, our analysis suggests the possibility of enhancing protein thermal stability by adequately redesigning the distribution of solvent-exposed, charged residues on the native protein surface. PMID- 10387060 TI - Rearrangement of domain elements of the Ca-ATPase in cardiac sarcoplasmic reticulum membranes upon phospholamban phosphorylation. AB - Phospholamban (PLB) is a major target of the beta-adrenergic cascade in the heart, and functions to modulate rate-limiting conformational transitions involving the transport activity of the Ca-ATPase. To investigate structural changes within the Ca-ATPase that result from the phosphorylation of PLB by cAMP dependent protein kinase (PKA), we have covalently bound the long-lived phosphorescent probe erythrosin isothiocyanate (Er-ITC) to cytoplasmic sequences within the Ca-ATPase. Under these labeling conditions, the Ca-ATPase remains catalytically active, indicating that observed changes in rotational dynamics reflect normal conformational transitions. Two major Er-ITC labeling sites were identified using electrospray ionization mass spectrometry (ESI-MS), corresponding to Lys464 and Lys650, which are respectively located within the phosphorylation and nucleotide binding domains of the Ca-ATPase. Frequency-domain phosphorescence measurements of the rotational dynamics of Er-ITC bound to these cytoplasmic sequences within the Ca-ATPase permit the resolution of the dynamic structure of individual domain elements relative to the overall rotational motion of the entire Ca-ATPase polypeptide chain. We observe a significant decrease in the rotational dynamics of Er-ITC bound to the Ca-ATPase upon phosphorylation of PLB by PKA, as evidenced by an increase in the residual anisotropy. These results suggest that phosphorylation of PLB results in a structural reorientation of the phosphorylation or nucleotide binding domains with respect to the membrane normal. In contrast, calcium activation of the Ca-ATPase in the presence of dephosphorylated PLB results in no detectable change in the rotational dynamics of Er-ITC, suggesting that calcium binding and PLB phosphorylation have distinct effects on the conformation of the Ca-ATPase. We suggest that PLB functions to alter the efficiency of phosphoenyzme formation following calcium activation of the Ca-ATPase by modulating the spatial arrangement between ATP bound in the nucleotide binding domain and Asp351 in the phosphorylation domain. PMID- 10387061 TI - Protein and nonprotein cysteinyl thiol modification by N-acetyl-p-benzoquinone imine via a novel ipso adduct. AB - N-acetyl-p-benzoquinone imine (NAPQI), a reactive metabolite of acetaminophen (APAP), can arylate and oxidize protein and nonprotein thiols in the pathogenesis of APAP-induced hepatotoxicity. We report the first direct evidence for the formation of a labile ipso adduct between glutathione (GSH) and NAPQI using a combination of techniques including liquid chromatography/tandem mass spectrometry and liquid chromatography/NMR spectroscopy. Decomposition kinetics of the GSH-NAPQI ipso adduct and product ratios suggested that the ipso adduct was readily reversible back to NAPQI under neutral and basic conditions. The significance of the ipso adduct is that it may migrate from its site of formation to other cell compartments where it can either oxidize protein thiols or covalently modify them. Ipso adduct formation with protein thiols was demonstrated with a cysteine protease, papain, whose catalytic activity relies on the presence of an active site cysteinyl thiol. The formation and reactions of cysteinyl thiol ipso adducts of NAPQI provides significant new insights into possible reactions of quinone imines with cellular peptides and proteins. PMID- 10387062 TI - Secondary structure extensions in Pyrococcus furiosus ferredoxin destabilize the disulfide bond relative to that in other hyperthermostable ferredoxins. Global consequences for the disulfide orientational heterogeneity. AB - The single cubane cluster ferredoxin (Fd) from the hyperthermophilic archaeon Pyrococcus furiosus (Pf) possesses several unique properties when compared even to Fds from other hyperthermophilic archaea or bacteria. These include an equilibrium molecular heterogeneity, a six- to seven-residue increase in size, an Asp rather than the Cys as one cluster ligand, and a readily reducible disulfide bond. NMR assignments and determination of both secondary structure and tertiary contacts remote from the paramagnetic oxidized cluster of Pf 3Fe Fd with an intact disulfide bond reported previously (Teng Q., Zhou, Z. H., Smith, E. T., Busse, S. C., Howard, J. B. Adams, M. W. W., and La Mar, G. (1994) Biochemistry 33, 6316-6328) are extended here to the 4Fe oxidized cluster WT (1H and 15N) and D14C (1H only) Fds with an intact disulfide bond and to the 4Fe oxidized WT Fd (1H and 15N) with a cleaved disulfide bond. All forms are shown to possess a long (13-member) alpha-helix, two beta-sheets (one double-, one triple-stranded), and three turns outside the cluster vicinity, each with tertiary contacts among themselves as found in other Fds. While the same secondary structural elements, with similar tertiary contacts, are found in other hyperthermostable Fds, Pf Fd has two elements, the long helix and the triple-stranded beta-sheet, that exhibit extensions and form multiple tertiary contacts. All Pf Fd forms with an intact disulfide bond exhibit a dynamic equilibrium heterogeneity which is shown to modulate a hydrogen-bonding network in the hydrophobic core that radiates from the Cys21-Cys48 disulfide bond and encompasses residues Lys36, Val24, Cys21, and Cys17 and the majority of the long helix. The heterogeneity is attributed to population of the alternate S and R chiralities of the disulfide bond, each destabilized by steric interactions with the extended alpha-helix. Comparison of the chemical shifts and their temperature gradients reveals that the molecular structure of the protein with the less stable R disulfide resembles that of the Fd with a cleaved disulfide bond. Both cluster architecture (3Fe vs 4Fe) and ligand mutation (Cys for Asp14) leave the disulfide orientational heterogeneity largely unperturbed. It is concluded that the six- to seven-residue extension that results in a longer helix and larger beta-sheet in Pf Fd, relative to other hyperthermostable Fds, more likely serves to destabilize the disulfide bond, and hence make it more readily reducible, than to significantly increase protein thermostability. PMID- 10387063 TI - Phosphate release during microtubule assembly: what stabilizes growing microtubules? AB - The molecular mechanism underlying microtubule dynamic instability depends on the relationship between the addition of tubulin-GTP to a growing microtubule and its hydrolysis in the microtubule lattice to tubulin-GDP, with release of inorganic phosphate (Pi). Since this relationship remains controversial, we have re examined the release of Pi upon microtubule assembly using a fluorometric assay for Pi, based on the phosphate-binding protein of Escherichia coli [Brune M., Hunter, J. L., Corrie, J. E. T., and Webb, M. R. (1994) Biochemistry 33, 8262 8271]. Microtubule assembly and Pi release were monitored simultaneously in a standard fluorimeter as an increase in the turbidity and fluorescence, respectively, in tubulin-GTP solutions assembled under conditions supporting dynamic instability. At the steady state of assembly, Pi release is nonlinear with respect to time, proceeding at a rate determined by the following: (a) the intrinsic GTPase activity of the nonpolymerized tubulin-GTP, and (b) the microtubule number concentration, which decreases progressively. Direct observation of the time course of nucleated microtubule assembly indicates that Pi release is closely coupled to microtubule elongation, even during the initial stages of assembly when uncoupling of tubulin-GTP addition and GTP hydrolysis would be most evident. Studies of the inhibition and reversal of the growth phase by cytostatic drugs show no evidence of a burst of Pi release. We conclude that nucleotide hydrolysis can keep pace with tubulin-GTP addition rates of 200 molecules per second per microtubule and that extended caps of tubulin-GTP or tubulin-GDP-Pi are not generated in normal assembly, nor are they required to stabilize growing microtubules or to support the phenomenon of dynamic instability of microtubules at the steady state. PMID- 10387064 TI - Carotenoid oxidation in photosystem II. AB - The oxidation of carotenoid upon illumination at low temperature has been studied in Mn-depleted photosystem II (PSII) using EPR and electronic absorption spectroscopy. Illumination of PSII at 20 K results in carotenoid cation radical (Car+*) formation in essentially all of the centers. When a sample which was preilluminated at 20 K was warmed in darkness to 120 K, Car+* was replaced by a chlorophyll cation radical. This suggests that carotenoid functions as an electron carrier between P680, the photooxidizable chlorophyll in PSII, and ChlZ, the monomeric chlorophyll which acts as a secondary electron donor under some conditions. By correlating with the absorption spectra at different temperatures, specific EPR signals from Car+* and ChlZ+* are distinguished in terms of their g values and widths. When cytochrome b559 (Cyt b559) is prereduced, illumination at 20 K results in the oxidation of Cyt b559 without the prior formation of a stable Car+*. Although these results can be reconciled with a linear pathway, they are more straightforwardly explained in terms of a branched electron-transfer pathway, where Car is a direct electron donor to P680(+), while Cyt b559 and ChlZ are both capable of donating electrons to Car+*, and where the ChlZ donates electrons when Cyt b559 is oxidized prior to illumination. These results have significant repercussions on the current thinking concerning the protective role of the Cyt b559/ChlZ electron-transfer pathways and on structural models of PSII. PMID- 10387065 TI - Ultraviolet resonance Raman examination of horse apomyoglobin acid unfolding intermediates. AB - We have used UV resonance Raman spectroscopy to study the acid-induced denaturation of horse apomyoglobin (apoMb) between pH 7. 0 and 1.8. The 206.5 nm excited Raman spectra are dominated by amide vibrations, which are used to quantitatively determine the apoMb secondary structure. The 229 nm excited Raman spectra are dominated by the Tyr and Trp Raman bands, which are analyzed to examine changes of Tyr and Trp environments and solvent exposures. We observe two partially unfolded apoMb intermediates at pH 4 and pH 2, while we observe only one partially unfolded holoMb intermediate at 2, in which the G and H helices are mainly intact, while the rest of protein is unfolded. This partially unfolded holoMb intermediate at pH 2 is essentially identical to the pH 2 apoMb intermediate. The partially unfolded pH 4 apoMb intermediate is composed of the three folded A, G, and H helices and contains 38% helical structure. The changes in the Trp Raman cross sections during the acid-induced denaturation indicates that Trp 7 is likely to be fully exposed in the apoMb pH 4 intermediate and that the A helix melts with a pKa approximately 3.5. PMID- 10387066 TI - An unexpected role for the active site base in cofactor orientation and flexibility in the copper amine oxidase from Hansenula polymorpha. AB - The role of the active site aspartate base in the aminotransferase mechanism of the copper amine oxidase from the yeast Hansenula polymorpha has been probed by site-directed mutagenesis. The D319E mutant catalyzes the oxidation of methylamine and phenethylamine, but not that of benzylamine. kcat/Km for methylamine is found to be 80-fold reduced compared to that of the wild type. Viscosogen and substrate and solvent deuteration have no effect on this parameter for D319E, which is suggestive of limitation of kcat/Km by a conformational change. This conformational change is proposed to be the movement of the cofactor into a productive orientation upon the binding of substrate. In the absence of substrate, a flipped cofactor orientation is likely, on the basis of resonance Raman evidence that the C5 carbonyl of the cofactor is less solvent accessible than the C3 hydrogen. kcat for D319E methylamine oxidase is reduced 200-fold compared to that of the wild type and is unaffected by substrate deuteration, but displays a substantial solvent isotope effect. A 428 nm absorbance is evident under conditions of saturating methylamine and oxygen with D319E. The D319N mutant is observed to produce a similar absorbance at 430 nm when treated with ammonia despite the fact that this mutant has no amine oxidase activity. Resonance Raman spectroscopy indicates the formation of a covalent ammonia adduct and identifies it as the deprotonated iminoquinone. In contrast, when the D319E mutant is reacted with ammonia, it gives predominantly a 340-350 nm species. This absorbance is ascribed to a localization of the cofactor oxyanion induced by binding of the cation at the active site and not to covalent adduct formation. Resonance Raman spectroscopic examination of the steady state species of D319E methylamine oxidation, in combination with the kinetic data, indicates that the 428 nm species is the deprotonated iminoquinone produced upon reoxidation of the reduced cofactor. A model is proposed in which a central role of the active site base is to position the free cofactor and several enzyme intermediates for optimal activity. PMID- 10387068 TI - Oxidized and reduced Azotobacter vinelandii ferredoxin I at 1.4 A resolution: conformational change of surface residues without significant change in the [3Fe 4S]+/0 cluster. AB - The refined structure of reduced Azotobacter vinelandii 7Fe ferredoxin FdI at 100 K and 1.4 A resolution is reported, permitting comparison of [3Fe-4S]+ and [3Fe 4S]0 clusters in the same protein at near atomic resolution. The reduced state of the [3Fe-4S]0 cluster is established by single-crystal EPR following data collection. Redundant structures are refined to establish the reproducibility and accuracy of the results for both oxidation states. The structure of the [4Fe 4S]2+ cluster in four independently determined FdI structures is the same within the range of derived standard uncertainties, providing an internal control on the experimental methods and the refinement results. The structures of the [3Fe-4S]+ and [3Fe-4S]0 clusters are also the same within experimental error, indicating that the protein may be enforcing an entatic state upon this cluster, facilitating electron-transfer reactions. The structure of the FdI [3Fe-4S]0 cluster allows direct comparison with the structure of a well-characterized [Fe3S4]0 synthetic analogue compound. The [3Fe-4S]0 cluster displays significant distortions with respect to the [Fe3S4]0 analogue, further suggesting that the observed [3Fe-4S]+/0 geometry in FdI may represent an entatic state. Comparison of oxidized and reduced FdI reveals conformational changes at the protein surface in response to reduction of the [3Fe-4S]+/0 cluster. The carboxyl group of Asp15 rotates approximately 90 degrees, Lys84, a residue hydrogen bonded to Asp15, adopts a single conformation, and additional H2O molecules become ordered. These structural changes imply a mechanism for H+ transfer to the [3Fe-4S]0 cluster in agreement with electrochemical and spectroscopic results. PMID- 10387067 TI - The active site base controls cofactor reactivity in Escherichia coli amine oxidase: x-ray crystallographic studies with mutational variants. AB - Amine oxidases utilize a proton abstraction mechanism following binding of the amine substrate to the C5 position of the cofactor, the quinone form of trihydroxyphenylalanine (TPQ). Previous work [Wilmot, C. M., et al. (1997) Biochemistry 36, 1608-1620] has shown that Asp383 in Escherichia coliamine oxidase (ECAO) is the catalytic base which performs the key step of proton abstraction. This paper explores in more depth this and other roles of Asp383. The crystal structures of three mutational variants are presented together with their catalytic properties, visible spectra, and binding properties for a substrate-like inhibitor, 2-hydrazinopyridine (2-HP), in comparison to those of the wild type enzyme. In wild type ECAO, the TPQ is located in a wedge-shaped pocket which allows more freedom of movement at the substrate binding position (C5) than for TPQ ring carbons C1-C4. A role of Asp383, whose carboxylate is located close to O5, is to stabilize the TPQ in its major conformation in the pocket. Replacement of Asp383 with the isostructural, but chemically distinct, Asn383 does not affect the location or dynamics of the TPQ cofactor significantly, but eliminates catalytic activity and drastically reduces the affinity for 2-HP. Removal of the side chain carboxyl moiety, as in Ala383, additionally allows the TPQ the greater conformational flexibility to coordinate to the copper, which demonstrates that Asp383 helps maintain the active site structure by preventing TPQ from migrating to the copper. Glu383 has a greatly decreased catalytic activity, as well as a decreased affinity for 2-HP relative to that of wild type ECAO. The electron density reveals that the longer side chain of Glu prevents the pivotal motion of the TPQ by hindering its movement within the wedge-shaped active site pocket. The results show that Asp383 performs multiple roles in the catalytic mechanism of ECAO, not only in acting as the active site base at different stages of the catalytic cycle but also in regulating the mobility of the TPQ that is essential to catalysis. PMID- 10387069 TI - Probing the unfolding region in a thermolysin-like protease by site-specific immobilization. AB - Protein stabilization by immobilization has been proposed to be most effective if the protein is attached to the carrier at that region where unfolding is initiated. To probe this hypothesis, we have studied the effects of site-specific immobilization on the thermal stability of mutants of the thermolysin-like protease from Bacillus stearothermophilus (TLP-ste). This enzyme was chosen because previous studies had revealed which parts of the molecule are likely to be involved in the early steps of thermal unfolding. Cysteine residues were introduced by site-directed mutagenesis into various positions of a cysteine-free variant of TLP-ste. The mutant enzymes were immobilized in a site-specific manner onto Activated Thiol-Sepharose. Two mutants (T56C, S65C) having their cysteine in the proposed unfolding region of TLP-ste showed a 9- and 12-fold increase in half lives at 75 degrees C due to immobilization. The stabilization by immobilization was even larger (33-fold) for the T56C/S65C double mutant enzyme. In contrast, mutants containing cysteines in other parts of the TLP-ste molecule (N181C, S218C, T299C) showed only small increases in half-lives due to immobilization (maximum 2.5-fold). Thus, the stabilization obtained by immobilization was strongly dependent on the site of attachment. It was largest when TLP-ste was fixed to the carrier through its postulated unfolding region. The concept of the unfolding region may be of general use for the design of strategies to stabilize proteins. PMID- 10387070 TI - Water-soluble beta-sheet models which self-assemble into fibrillar structures. AB - Self-assembly of beta-sheet domains resulting in the formation of pathogenic, fibrillar protein aggregates (amyloids) is a characteristic feature of various medical disorders. These include neurodegenerative diseases, such as Alzheimer's, Huntington's, and Creutzfeldt-Jacob's. A significant problem in studying such aggregation processes is the poor solubility of these beta-sheet complexes. The present work describes water-soluble de novo beta-sheet peptides which self assemble into fibrillar structures. The model peptides enable studies of the relationship between beta-sheet stability and association behavior. The peptides [DPKGDPKG-(VT)n-GKGDPKPD-NH2, n = 3-8] are composed of a central beta-sheet forming domain (VT-sequence), and N- and C-terminal nonstructured octapeptide sequences which promote water solubility. Conformational analyses by circular dichroism and Fourier transform infrared spectroscopy indicate the influence of peptide length, D-amino acid substitution, and concentration on the ability of the peptides to form stable beta-sheet structures. The association behavior investigated by analytical ultracentrifugation and dynamic light scattering was found to correlate strongly with the stability of a beta-sheet conformation. Model peptides with n >/= 6 form stable, water-soluble beta-sheet complexes with molecular masses of more than 2000 kDa, which are organized in fibrillar structures. The fibrils examined by Congo Red staining and electron microscopy show some similarities with naturally occurring amyloid fibrils. PMID- 10387071 TI - Calculated protein and proton motions coupled to electron transfer: electron transfer from QA- to QB in bacterial photosynthetic reaction centers. AB - Reaction centers from Rhodobacter sphaeroides were subjected to Monte Carlo sampling to determine the Boltzmann distribution of side-chain ionization states and positions and buried water orientation and site occupancy. Changing the oxidation states of the bacteriochlorophyll dimer electron donor (P) and primary (QA) and secondary (QB) quinone electron acceptors allows preparation of the ground (all neutral), P+QA-, P+QB-, P0QA-, and P0QB- states. The calculated proton binding going from ground to other oxidation states and the free energy of electron transfer from QA-QB to form QAQB- (DeltaGAB) compare well with experiment from pH 5 to pH 11. At pH 7 DeltaGAB is measured as -65 meV and calculated to be -80 meV. With fixed protein positions as in standard electrostatic calculations, DeltaGAB is +170 meV. At pH 7 approximately 0.2 H+/protein is bound on QA reduction. On electron transfer to QB there is little additional proton uptake, but shifts in side chain protonation and position occur throughout the protein. Waters in channels leading from QB to the surface change site occupancy and orientation. A cluster of acids (GluL212, AspL210, and L213) and SerL223 near QB play important roles. A simplified view shows this cluster with a single negative charge (on AspL213 with a hydrogen bond to SerL233) in the ground state. In the QB- state the cluster still has one negative charge, now on the more distant AspL210. AspL213 and SerL223 move so SerL223 can hydrogen bond to QB-. These rearrangements plus other changes throughout the protein make the reaction energetically favorable. PMID- 10387072 TI - Cloning, sequence analysis, and expression of active Phrixothrix railroad-worms luciferases: relationship between bioluminescence spectra and primary structures. AB - Phrixothrix railroad-worms emit yellow-green light through 11 pairs of lateral lanterns along the body and red light through two cephalic lanterns. The cDNAs for the lateral lanterns luciferase of Phrixothrix vivianii, which emit green light (lambda max= 542 nm), and for the head lanterns of P. hirtus, which emit the most red-shifted bioluminescence (lambda max= 628 nm) among luminescent beetles, were cloned. Positive clones which emitted green (PvGR: lambda max= 549 nm) and red (PhRE: lambda max= 622 nm) bioluminescence were isolated. The lucifereases coded by PvGR (545 amino acid residues) and PhRE (546 amino acid residues) cDNAs share 71% identity. PvGR and PhRE luciferases showed 50-55% and 46-49% identity with firefly luciferases, respectively, and 47-49% with click beetle luciferases. PhRE luciferase has some unique residues which replace invariant residues in other beetle luciferases. The additional residue Arg 352 in PhRE, which is deleted in PvGR polypeptide, seems to be another important structural feature associated with red light production. As in the case of other railroad-worms and click-beetle luciferases studied, Phrixothrix luciferases do not undergo the typical red shift suffered by firefly luciferases upon decreasing pH, a property which might be related to the many amino acid residues shared in common between railroad-worm and click-beetle luciferase. PMID- 10387073 TI - Divalent cations stabilize the alpha 1 beta 1 integrin I domain. AB - Recent structural and functional analyses of alpha integrin subunit I domains implicate a region in cation and ligand binding referred to as the metal ion dependent adhesion site (MIDAS). Although the molecular interactions between Mn2+ and Mg2+ and the MIDAS region have been defined by crystallographic analyses, the role of cation in I domain function is not well understood. Recombinant alpha 1 beta 1 integrin I domain (alpha1-I domain) binds collagen in a cation-dependent manner. We have generated and characterized a panel of antibodies directed against the alpha1-I domain, and selected one (AJH10) that blocks alpha 1 beta 1 integrin function for further study. The epitope of AJH10 was localized within the loop between the alpha 3 and alpha 4 helices which contributes one of the metal coordination sites of the MIDAS structure. Kinetic analyses of antibody binding to the I domain demonstrate that divalent cation is required to stabilize the epitope. Denaturation experiments demonstrate that cation has a dramatic effect on the stabilization of the I domain structure. Mn2+ shifts the point at which the I domain denatures from 3.4 to 6.3 M urea in the presence of the denaturant, and from 49.5 to 58.6 degrees C following thermal denaturation. The structural stability provided to the alpha1-I domain by divalent cations may contribute to augmented ligand binding that occurs in the presence of these cations. PMID- 10387074 TI - Binding of cardiac troponin-I147-163 induces a structural opening in human cardiac troponin-C. AB - The interaction of troponin-C (TnC) with troponin-I (TnI) plays a central role in skeletal and cardiac muscle contraction. We have recently shown that the binding of Ca2+ to cardiac TnC (cTnC) does not induce an "opening" of the regulatory domain in order to interact with cTnI [Sia, S. K., et al. (1997) J. Biol. Chem. 272, 18216-18221; Spyracopoulos et al. (1997) Biochemistry 36, 12138-12146], which is in contrast to the regulatory N-domain of skeletal TnC (sTnC). This implies that the mode of interaction between cTnC and cTnI may be different than that between sTnC and sTnI. In sTnI, a region downstream from the inhibitory region (residues 115-131) has been shown to bind the exposed hydrophobic pocket of Ca2+-saturated sNTnC [McKay, R. T., et al. (1997) J. Biol. Chem. 272, 28494 28500]. The present study demonstrates that the corresponding region in cTnI (residues 147-163) binds to the regulatory domain of cTnC only in the Ca2+ saturated state to form a 1:1 complex, with an affinity approximately six times weaker than that between the skeletal counterparts. Thus, while Ca2+ does not cause opening, it is required for muscle regulation. The solution structure of the cNTnC.Ca2+.cTnI147-163 complex has been determined by multinuclear multidimensional NMR spectroscopy. The structure reveals an open conformation for cNTnC, similar to that of Ca2+-saturated sNTnC. The bound peptide adopts a alpha helical conformation spanning residues 150-157. The C-terminus of the peptide is unstructured. The open conformation for Ca2+-saturated cNTnC in the presence of cTnI (residues 147-163) accommodates hydrophobic interactions between side chains of the peptide and side chains at the interface of A and B helices of cNTnC. Thus the mechanistic differences between the regulation of cardiac and skeletal muscle contraction can be understood in terms of different thermodynamics and kinetics equilibria between essentially the same structure states. PMID- 10387075 TI - The HELLGH motif of rat liver dipeptidyl peptidase III is involved in zinc coordination and the catalytic activity of the enzyme. AB - The role of the HELLGH (residues 450-455) motif in the sequence of rat dipeptidyl peptidase III (EC 3.4.14.4) was investigated by replacing Glu451 with an alanine or an aspartic acid residue and by replacing His450 and His455 with a tyrosine residue by site-directed mutagenesis. Mutated cDNAs were expressed three or four times in Escherichia coli, and the resulting proteins were purified to apparent homogeneity. None of the expressed mutated proteins exhibited DPP III activity. The mutants of Glu451 contained 1 mol of zinc per mole of protein, but mutants His450 and His455 did not contain significant amounts of zinc as determined by atomic absorption spectrometry. The Leu453-deleted enzyme (having the zinc aminopeptidase motif HExxH-18-E) had almost the same order of binding affinity (for Arg-Arg-2-naphthylamide) as the wild-type enzyme, but the specificity constant was about 10%. These results provide evidence that the suitable number of amino acids included between Glu451 and His455 is three residues for the enzyme activity and confirm that residues His450, His455, and Glu451 are involved in zinc coordination and catalytic activity. PMID- 10387076 TI - Solution structure of the N-terminal F1 module pair from human fibronectin. AB - Multiple sites within the N-terminal domain (1-5F1) of fibronectin have been implicated previously in fibronectin matrix assembly, heparin binding, and binding to cell surface proteins of pathogenic bacteria. The solution structure of 1F1(2)F1, the N-terminal F1 module pair from human fibronectin, has been determined using NMR spectroscopy. Both modules in the pair conform to the F1 consensus fold. In 4F1(5)F1, the only other F1 module pair structure available, there is a well-defined intermodule interface; in 1F1(2)F1, however, there is no detectable interface between the modules. Comparison of the backbone 15N-{1H} NOE values for both module pairs confirms that the longer intermodule sequence in 1F1(2)F1 is flexible and that the stabilization of the 4F1 C-D loop observed in 4F1(5)F1, as a result of the intermodule interface, is not observed in 1F1(2)F1. PMID- 10387078 TI - Crystal structure of S-adenosylhomocysteine hydrolase from rat liver. AB - The crystal structure of rat liver S-adenosyl-L-homocysteine hydrolase (AdoHcyase, EC 3.3.1.1) which catalyzes the reversible hydrolysis of S adenosylhomocysteine (AdoHcy) has been determined at 2.8 A resolution. AdoHcyase from rat liver is a tetrameric enzyme with 431 amino acid residues in each identical subunit. The subunit is composed of the catalytic domain, the NAD+ binding domain, and the small C-terminal domain. Both catalytic and NAD+-binding domains are folded into an ellipsoid with a typical alpha/beta twisted open sheet structure. The C-terminal section is far from the main body of the subunit and extends into the opposite subunit. An NAD+ molecule binds to the consensus NAD+ binding cleft of the NAD+-binding domain. The peptide folding pattern of the catalytic domain is quite similar to the patterns observed in many methyltransferases. Although the crystal structure does not contain AdoHcy or its analogue, there is a well-formed AdoHcy-binding crevice in the catalytic domain. Without introducing any major structural changes, an AdoHcy molecule can be placed in the catalytic domain. In the structure described here, the catalytic and NAD+-binding domains are quite far apart from each other. Thus, the enzyme appears to have an "open" conformation in the absence of substrate. It is likely that binding of AdoHcy induces a large conformational change so as to place the ribose moiety of AdoHcy in close proximity to the nicotinamide moiety of NAD+. A catalytic mechanism of AdoHcyase has been proposed on the basis of this crystal structure. Glu155 acts as a proton acceptor from the O3'-H when the proton of C3' H is abstracted by NAD+. His54 or Asp130 acts as a general acid-base catalyst, while Cys194 modulates the oxidation state of the bound NAD+. The polypeptide folding pattern of the catalytic domain suggests that AdoHcy molecules can travel freely to and from AdoHcyase and methyltransferases to properly regulate methyltransferase activities. We believe that the crystal structure described here can provide insight into the molecular architecture of this important regulatory enzyme. PMID- 10387077 TI - Solution structures of the C-terminal domain of cardiac troponin C free and bound to the N-terminal domain of cardiac troponin I. AB - The N-terminal domain of cardiac troponin I (cTnI) comprising residues 33-80 and lacking the cardiac-specific amino terminus forms a stable binary complex with the C-terminal domain of cardiac troponin C (cTnC) comprising residues 81-161. We have utilized heteronuclear multidimensional NMR to assign the backbone and side chain resonances of Ca2+-saturated cTnC(81-161) both free and bound to cTnI(33 80). No significant differences were observed between secondary structural elements determined for free and cTnI(33-80)-bound cTnC(81-161). We have determined solution structures of Ca2+-saturated cTnC(81-161) free and bound to cTnI(33-80). While the tertiary structure of cTnC(81-161) is qualitatively similar to that observed free in solution, the binding of cTnI(33-80) results mainly in an opening of the structure and movement of the loop region between helices F and G. Together, these movements provide the binding site for the N terminal domain of cTnI. The putative binding site for cTnI(33-80) was determined by mapping amide proton and nitrogen chemical shift changes, induced by the binding of cTnI(33-80), onto the C-terminal cTnC structure. The binding interface for cTnI(33-80), as suggested from chemical shift changes, involves predominantly hydrophobic interactions located in the expanded hydrophobic pocket. The largest chemical shift changes were observed in the loop region connecting helices F and G. Inspection of available TnC sequences reveals that these residues are highly conserved, suggesting a common binding motif for the Ca2+/Mg2+-dependent interaction site in the TnC/TnI complex. PMID- 10387079 TI - Investigation of the local structure and dynamics of the H subunit of the mitochondrial glycine decarboxylase using heteronuclear NMR spectroscopy. AB - The lipoate-dependent H protein plays a pivotal role in the catalytic cycle of the glycine decarboxylase complex (GDC), undergoing reducing methylamination, methylene transfer, and oxidation. The local structure and backbone dynamics of the methylamine-loaded H (Hmet), oxidized H (Hox), and H apoprotein (Hapo) have been investigated in solution. Filtered NOESY experiments using a [13C]Hmet as well as comparison of the heteronuclear shifts between the Hox and Hmet proteins demonstrate that the methylamine group is located inside a cleft of the protein. Furthermore, this group appears to be locked in this configuration as indicated by the high value of the activation energy (37 kcal/mol) of the global unloading reaction and by its restricted mobility, deduced from 13C relaxation measurements. Comparisons of the 1H and 15N chemical shifts and 15N relaxation in the three forms suggest that part of the lipoyl-lysine arm interacts with the protein polypeptide in the Hox and Hmet. The major change induced by the loading of the methylamine group concerns the C-terminal helix whose mobility becomes completely restricted compared to those of the Hox and Hapo. This C-terminal helix exhibits different reorientational characteristics in the three forms, which can be explained in the Hapo by a model consisting of a twisting motion about an axis passing through the helix. Our results indicate that the model of a freely swinging arm proposed for other lipoate-containing proteins is not acceptable in solution for the GDC. The implication of this observation in terms of the mechanism of the interaction of the H protein with the T protein, its physiological partner during the catalytic cycle, is discussed. PMID- 10387080 TI - Crystal structure of rabbit cytosolic serine hydroxymethyltransferase at 2.8 A resolution: mechanistic implications. AB - Serine hydroxymethyltransferase (SHMT) catalyzes the reversible cleavage of serine to form glycine and single carbon groups that are essential for many biosynthetic pathways. SHMT requires both pyridoxal phosphate (PLP) and tetrahydropteroylpolyglutamate (H4PteGlun) as cofactors, the latter as a carrier of the single carbon group. We describe here the crystal structure at 2.8 A resolution of rabbit cytosolic SHMT (rcSHMT) in two forms: one with the PLP covalently bound as an aldimine to the Nepsilon-amino group of the active site lysine and the other with the aldimine reduced to a secondary amine. The rcSHMT structure closely resembles the structure of human SHMT, confirming its similarity to the alpha-class of PLP enzymes. The structures reported here further permit identification of changes in the PLP group that accompany formation of the geminal diamine complex, the first intermediate in the reaction pathway. On the basis of the current mechanism derived from solution studies and the properties of site mutants, we are able to model the binding of both the serine substrate and the H4PteGlun cofactor. This model explains the properties of several site mutants of SHMT and offers testable hypotheses for a more detailed mechanism of this enzyme. PMID- 10387081 TI - Nonaggregating mutant of recombinant human hexokinase I exhibits wild-type kinetics and rod-like conformations in solution. AB - Hexokinase I governs the rate-limiting step of glycolysis in brain tissue, being inhibited by its product, glucose 6-phosphate, and allosterically relieved of product inhibition by phosphate. On the basis of small-angle X-ray scattering, the wild-type enzyme is a monomer in the presence of glucose and phosphate at protein concentrations up to 10 mg/mL, but in the presence of glucose 6 phosphate, is a dimer down to protein concentrations as low as 1 mg/mL. A mutant form of hexokinase I, specifically engineered by directed mutation to block dimerization, remains monomeric at high protein concentration under all conditions of ligation. This nondimerizing mutant exhibits wild-type activity, potent inhibition by glucose 6-phosphate, and phosphate reversal of product inhibition. Small-angle X-ray scattering data from the mutant hexokinase I in the presence of glucose/phosphate, glucose/glucose 6-phosphate, and glucose/ADP/Mg2+/AlF3 are consistent with a rodlike conformation for the monomer similar to that observed in crystal structures of the hexokinase I dimer. Hence, any mechanism for allosteric regulation of hexokinase I should maintain a global conformation of the polypeptide similar to that observed in crystallographic structures. PMID- 10387082 TI - NMR structure and functional studies of the Mu repressor DNA-binding domain. AB - The repressor protein of bacteriophage Mu establishes and maintains lysogeny by shutting down transposition functions needed for phage DNA replication. It interacts with several repeated DNA sequences within the early operator, preventing transcription from two divergent promoters. It also directly represses transposition by competing with the MuA transposase for an internal activation sequence (IAS) that is coincident with the operator and required for efficient transposition. The transposase and repressor proteins compete for the operator/IAS region using homologous DNA-binding domains located at their amino termini. Here we present the solution structure of the amino-terminal DNA-binding domain from the repressor protein determined by heteronuclear multidimensional nuclear magnetic resonance spectroscopy. The structure of the repressor DNA binding domain provides insights into the molecular basis of several temperature sensitive mutations and, in combination with complementary experiments using flourescence anisotropy, surface plasmon resonance, and circular dichroism, defines the structural and biochemical differences between the transposase and repressor DNA-binding modules. We find that the repressor and enhancer domains possess similar three-dimensional structures, thermostabilities, and intrinsic affinities for DNA. This latter result suggests that the higher affinity of the full-length repressor relative to that of the MuA transposase protein originates from cooperative interactions between repressor protomers and not from intrinsic differences in their DNA-binding domains. In addition, we present the results of nucleotide and amino acid mutagenesis which delimits the minimal repressor DNA binding module and coarsely defines the nucleotide dependence of repressor binding. PMID- 10387083 TI - Effects of glycosylation on the structure and dynamics of eel calcitonin in micelles and lipid bilayers determined by nuclear magnetic resonance spectroscopy. AB - The three-dimensional structures of eel calcitonin (CT) and two glycosylated CT derivatives, [Asn(GlcNAc)3]-CT (CT-GlcNAc) and [Asn(Man6-GlcNAc2)3]-CT (CT-M6), in micelles were determined by solution NMR spectroscopy. The topologies of these peptides associated with oriented lipid bilayers were determined with solid-state NMR. All of the peptides were found to have an identical conformation in micelles characterized by an amphipathic alpha-helix consisting of residues Ser5 through Leu19 followed by an unstructured region at the C-terminus. The overall conformation of the peptide moiety was not affected by the glycosylation. Nevertheless, comparison of the relative exchange rates of the Leu12 amide proton might suggest the possibility that fluctuations of the alpha-helix are reduced by glycosylation. The presence of NOEs between the carbohydrate and the peptide moieties of CT-GlcNAc and CT-M6 and the amide proton chemical shift data suggested that the carbohydrate interacted with the peptide, and this might account for the conformational stabilization of the alpha-helix. Both the unmodified CT and the glycosylated CT were found to have orientations with their helix axes parallel to the plane of the lipid bilayers by solid-state NMR spectroscopy. PMID- 10387084 TI - X-ray structure of Novamyl, the five-domain "maltogenic" alpha-amylase from Bacillus stearothermophilus: maltose and acarbose complexes at 1.7A resolution. AB - The three-dimensional structure of the Bacillus stearothermophilus "maltogenic" alpha-amylase, Novamyl, has been determined by X-ray crystallography at a resolution of 1.7 A. Unlike conventional alpha-amylases from glycoside hydrolase family 13, Novamyl exhibits the five-domain structure more usually associated with cyclodextrin glycosyltransferase. Complexes of the enzyme with both maltose and the inhibitor acarbose have been characterized. In the maltose complex, two molecules of maltose are found in the -1 to -2 and +2 to +3 subsites of the active site, with two more on the C and E domains. The C-domain maltose occupies a position identical to one previously observed in the Bacillus circulans CGTase structure [Lawson, C. L., et al. (1994) J. Mol. Biol. 236, 590-600], suggesting that the C-domain plays a genuine biological role in saccharide binding. In the acarbose-maltose complex, the tetrasaccharide inhibitor acarbose is found as an extended hexasaccharide species, bound in the -3 to +3 subsites. The transition state mimicking pseudosaccharide is bound in the -1 subsite of the enzyme in a 2H3 half-chair conformation, as expected. The active site of Novamyl lies in an open gully, fully consistent with its ability to perform internal cleavage via an endo as opposed to an exo activity. PMID- 10387085 TI - Orientation of the headgroup of phosphatidylinositol in a model biomembrane as determined by neutron diffraction. AB - Derivatives of the sodium salt of dimyristoylphosphatidylinositol (DMPI) have been synthesized specifically deuterated in the headgroup. A 50:50 (molar) mixture of DMPI with dimyristoylphosphatidylcholine (DMPC) hydrated to the level of 16 waters/lipid gives a biomembrane-like Lalpha phase at 50 degrees C. Comparison of the neutron diffraction scattering profiles for deuterated and undeuterated membranes allowed the depth of each deuterium (hydrogen) within the bilayer to be determined to +/-0.5 A. This gave the orientation of the inositol ring which lies more-or-less along the bilayer normal projecting directly out into the water. This orientation is similar to that of the sugar residue in glycolipids and confirms previous models for PI. On the assumption that the (P)O DAG bond is more-or-less parallel to the bilayer normal, it is consistent with a roughly trans, trans, trans, gauche- conformation for the glyceryl-phosphate inositol link. In the case of DMPI, it is the C4-hydroxy group which is most fully extended into the water layer, but when this is phosphorylated, the inositol ring turns over and tilts so that the C5-hydroxy group is now the one furthest extended into the water layer. Hence, at each stage in the pathway PI - > PI-4P --> PI-4,5-P2, it is the hydroxy position most exposed to the water which undergoes phosphorylation. Whereas the orientation of the inositol ring in DMPI can be seen simply as maximizing its hydration, the tilt of the ring in DMPI-4P cannot be explained in this way. It is suggested that it is due to an electrostatic interaction. PMID- 10387086 TI - Interaction kinetics of tetramethylrhodamine transferrin with human transferrin receptor studied by fluorescence correlation spectroscopy. AB - We applied fluorescence correlation spectroscopy (FCS) to characterize the interaction dynamics of fluorescence-labeled transferrin with transferrin receptor (hTfR) associates isolated from human placenta. The dissociation constant for the equilibrium binding of TMR-labeled ferri-transferrin to hTfR in detergent free solution was determined to be 7 +/- 3 nM. Binding curves were compatible with equal and independent binding sites present on the hTfR associates. Under pseudo-first-order conditions, with respect to transferrin, complex formation is monophasic. From these curves, association and dissociation rate constants for a reversible bimolecular binding reaction were determined, with (1.1 +/- 0.1) x 10(4) M-1 s-1 for the former and (6 +/- 4) x 10(-)4 s-1 for the latter. In dissociation exchange experiments, biphasic curves and concentration-independent reciprocal relaxation times were determined. From isothermal titration calorimetry experiments, we obtained an enthalpy change of 44.4 kJ/mol associated with the reaction. We thus conclude that the reaction is mainly enthalpy driven. PMID- 10387087 TI - Enthalpy and heat capacity changes for formation of an oligomeric DNA duplex: interpretation in terms of coupled processes of formation and association of single-stranded helices. AB - The thermodynamics of self-assembly of a 14 base pair DNA double helix from complementary strands have been investigated by titration (ITC) and differential scanning (DSC) calorimetry, in conjunction with van't Hoff analysis of UV thermal scans of individual strands. These studies demonstrate that thermodynamic characterization of the temperature-dependent contributions of coupled conformational equilibria in the individual "denatured" strands and in the duplex is essential to understand the origins of duplex stability and to derive stability prediction schemes of general applicability. ITC studies of strand association at 293 K and 120 mM Na+ yield an enthalpy change of -73 +/- 2 kcal (mol of duplex)-1. ITC studies between 282 and 312 K at 20, 50, and 120 mM Na+ show that the enthalpy of duplex formation is only weakly salt concentration dependent but is very strongly temperature-dependent, decreasing approximately linearly with increasing temperature with a heat capacity change (282-312 K) of 1.3 +/- 0.1 kcal K-1 (mol of duplex)-1. From DSC denaturation studies in 120 mM Na+, we obtain an enthalpy of duplex formation of -120 +/- 5 kcal (mol of duplex) 1 and an estimate of the corresponding heat capacity change of -0.8 +/- 0.4 kcal K-1 (mol of duplex)-1 at the Tm of 339 K. van't Hoff analysis of UV thermal scans on the individual strands indicates that single helix formation is noncooperative with a temperature-independent enthalpy change of -5.5 +/- 0.5 kcal at 120 mM Na+. From these observed enthalpy and heat capacity changes, we obtain the corresponding thermodynamic quantities for two fundamental processes: (i) formation of single helices from disordered strands, involving only intrastrand (vertical) interactions between neighboring bases; and (ii) formation of double helices by association (docking) of single helical strands, involving interstrand (horizontal and vertical) interactions. At 293 K and 120 mM Na+, we calculate that the enthalpy change for association of single helical strands is approximately -64 kcal (mol of duplex)-1 as compared to -210 kcal (mol of duplex) 1 calculated for duplex formation from completely unstructured single strands and to the experimental ITC value of -73 kcal (mol of duplex)-1. The intrinsic heat capacity change for association of single helical strands to form the duplex is found to be small and positive [ approximately 0.1 kcal K-1 (mol of duplex)-1], in agreement with the result of a surface area analysis, which also predicts an undetectably small heat capacity change for single helix formation. PMID- 10387088 TI - hSWI/SNF disrupts interactions between the H2A N-terminal tail and nucleosomal DNA. AB - We have employed a site-specific core histone-DNA cross-linking approach to investigate the mechanism of hSWI/SNF remodeling of a nucleosome. Remodeling results in the complete loss of canonical contacts between the N-terminal tail of H2A and DNA while new interactions are detected between this domain and DNA near the center of the original nucleosome. The data are consistent with a model in which remodeling results in the unraveling of a region of DNA from the edge of the nucleosome, leading to a repositioning of the H2A/H2B dimer to a noncanonical position near the center of the remodeled complex. Additionally, we find that prior cross-linking of the H2A N-terminal region to nucleosomal DNA does not restrict hSWI/SNF remodeling of the remainder of the nucleosome. Thus, disruption of both H2A-DNA interactions near the edge of the nucleosome is not an obligatory step in remodeling of the remainder of the complex. PMID- 10387089 TI - Divalent metal dependence of site-specific DNA binding by EcoRV endonuclease. AB - Measurements of binding equilibria of EcoRV endonuclease to DNA, for a series of base-analogue substrates, demonstrate that expression of sequence selectivity is strongly enhanced by the presence of Ca2+ ions. Binding constants were determined for short duplex oligodeoxynucleotides containing the cognate DNA site, three cleavable noncognate sites, and a fully nonspecific site. At pH 7.5 and 100 mM NaCl, the full range of specificity from the specific (tightest binding) to nonspecific (weakest binding) sites is 0.9 kcal/mol in the absence of metal ions and 5.8 kcal/mol in the presence of Ca2+. Precise determination of binding affinities in the presence of the active Mg2+ cofactor was found to be possible for substrates retaining up to 1.6% of wild-type activity, as determined by the rate of phosphoryl transfer. These measurements show that Ca2+ is a near-perfect analogue for Mg2+ in binding reactions of the wild-type enzyme with DNA base analogue substrates, as it provides identical DeltaDeltaG degrees bind values among the cleavable noncognate sites. Equilibrium dissociation constants of wild type and base-analogue sites were also measured for the weakly active EcoRV mutant K38A, in the presence of either Mg2+ or Ca2+. In this case, Ca2+ allows expression of a greater degree of specificity than does Mg2+. DeltaDeltaG degrees bind values of K38A toward specific versus nonspecific sites are 6.1 kcal/mol with Ca2+ and 3.9 kcal/mol with Mg2+, perhaps reflecting metal-specific conformational changes in the ground-state ternary complexes. The enhancement of binding specificity provided by divalent metal ions is likely to be general to many restriction endonucleases and other metal-dependent nucleic acid-modifying enzymes. These results strongly suggest that measurements of DNA binding affinities for EcoRV, and likely for many other restriction endonucleases, should be performed in the presence of divalent metal ions. PMID- 10387090 TI - Structure-activity analysis of the effects of lysophosphatidic acid on platelet aggregation. AB - Lysophosphatidic acid (1-acyl-sn-glycero-3-phosphate or LPA) is a phospholipid mediator displaying numerous and widespread biological activities and thought to act via G-protein-coupled receptors. Here we have studied the effects on human platelets of a number of LPA analogues, including two enantiomers of both N palmitoyl-(L)-serine-3-phosphate ((L) and (D)NAPS for N-acyl-phosphoserine) and 2 (R)-N-palmitoyl-norleucinol-1-phosphate ((R) and (S)PNPA), cyclic analogues of 1 acyl-sn-glycero-3-phosphate (cPA) and of 1-O-hexadecyl-sn-glycero-3-phosphate (cAGP), sphingosine-1-phosphate (SPP), as well as two palmitoyl derivatives of dioxazaphosphocanes bearing either a P-H or a P-OH bond (DOXP-H and DOXP-OH, respectively). Nine of these compounds induced platelet aggregation with the following order of potency: SPP < cAGP < DOXP-OH < (L)NAPS = (D)NAPS < (R)PNPA = (S)PNPA < LPA < AGP, EC50 varying between 9.8 nM and 8.3 microM. Two of these compounds (SPP and cAGP) appeared as weak agonists inducing platelet aggregation to only 33% and 41%, respectively, of the maximal response attained with LPA and other analogues. In cross-desensitization experiments, all of these compounds specifically inhibited LPA-induced aggregation, suggesting that they were all acting on the same receptor(s). In contrast, cPA and DOXP-H did not trigger platelet aggregation but instead specifically inhibited the effects of LPA in a concentration-dependent manner. The inhibitory action of cPA did not vary with the acyl chain length or the presence of a double bond and did not involve an increase in cAMP. These data thus confirm the lack of stereospecificity of platelet LPA receptor(s). In addition, since the order of potency of some analogues is different from that described in other cells, our results suggest that platelets contain (a) pharmacologically distinct receptor(s) whose molecular identity still remains to be established. Finally, this unique series of compounds might be used for further characterization of other endogenous or recombinant LPA receptors. PMID- 10387091 TI - Inhibition of neutrophil cathepsin G by oxidized mucus proteinase inhibitor. Effect of heparin. AB - Oxidation of mucus proteinase inhibitor (MPI) transforms Met73, the P'1 residue of its active center into methionine sulfoxide and lowers its affinity for neutrophil elastase [Boudier, C., and Bieth, J. G. (1994) Biochem. J. 303, 61 68]. Here, we show that the oxidized inhibitor has also a decreased affinity for neutrophil cathepsin G and pancreatic chymotrypsin. The Ki of the oxidized MPI cathepsin G complex (1.2 microM) is probably too high to be compatible with significant inhibition of cathepsin G in inflammatory lung secretions. Stopped flow kinetics shows that, within the inhibitor concentration range used, the mechanism of inhibition of cathepsin G and chymotrypsin by oxidized MPI is consistent with a one-step reaction, [equation in text] whereas the inhibition of elastase takes place in two steps, [equation in text]. Heparin, which accelerates the inhibition of the three proteinases by native MPI, also favors their interaction with oxidized MPI. Flow calorimetry shows that heparin binds oxidized MPI with Kd, Delta H degrees, and Delta S degrees values close to those reported for native MPI. In the presence of heparin, oxidized MPI inhibits cathepsin G via a two-step reaction characterized by Ki = 0.22 microM, k2 = 0.1 s-1, k-2 = 0.023 s-1, and Ki = 42 nM. Under these conditions, in vivo inhibition of cathepsin G is again possible. Heparin also improves the inhibition of chymotrypsin and elastase by oxidized MPI by increasing their kass or k2/Ki and decreasing their Ki. Our data suggest that oxidation of MPI during chronic bronchitis may lead to cathepsin G-mediated lung tissue degradation and that heparin may be a useful adjuvant of MPI-based therapy of acute lung inflammation in cystic fibrosis. PMID- 10387092 TI - Divalent cations stimulate preferential recognition of a viral DNA end by HIV-1 integrase. AB - In the presence of a divalent metal cofactor (Mg2+ or Mn2+), retroviral-encoded integrase (IN) catalyzes two distinct reactions: site-specific cleavage of two nucleotides from both 3' ends of viral DNA, and sequence-independent joining of the recessed viral ends to staggered phosphates in a target DNA. Here we investigate human immunodeficiency virus type 1 (HIV-1) IN-DNA interactions using surface plasmon resonance. The results show that IN forms tight complexes both with duplex oligonucleotides that represent the viral DNA ends and with duplex oligonucleotides with an unrelated sequence that represent a target DNA substrate. The IN-DNA complexes are stable in 4.0 M NaCl, or 50% (v/v) methanol, but they are not resistant to low concentrations of SDS, indicating that their stability is highly dependent on structural features of the protein. Divalent metal cofactors exert two distinct effects on the IN-DNA interaction. Mn2+ inhibits IN binding to a model target DNA with the apparent Kd increasing approximately 3-fold in the presence of this cation. On the other hand, Mn2+ (or Mg2+) stimulates the binding of IN to a model viral DNA end, decreasing the apparent Kd of this IN-viral DNA complex approximately 6-fold. Such metal mediated stimulation of the binding of IN to the viral DNA is totally abolished by substitution of the subterminal conserved CA/GT bp with a GT/CA bp, and is greatly diminished when the viral DNA end is recessed or "pre-processed." IN binds to a viral duplex oligonucleotide whose end was extended with nonviral sequences with kinetics similar to the nonviral model target DNA. This suggests that IN can distinguish the integrated DNA product from the viral donor DNA in the presence of divalent metal ion. Thus, our results show that preferential recognition of viral DNA by HIV-1 IN is achieved only in the presence of metal cofactor, and requires a free, wild-type viral DNA end. PMID- 10387093 TI - Characterization and crystallization of soluble human Fc gamma receptor II (CD32) isoforms produced in insect cells. AB - Fc gamma RII (CD32), the receptor for the Fc part of IgG, is responsible for the clearance of immunocomplexes by macrophages and plays a role in the regulation of antibody production by B cells. To investigate the process of immunocomplex binding in terms of stoichiometry and stability of the Fc gamma RII:IgG complex, we produced both Fc gamma RII isoforms (Fc gamma RIIa and Fc gamma RIIb) as soluble proteins in insect cells. The expressed proteins could be purified in high yields and were biologically active as judged by their ability to bind IgG. Thus, the minor glycosylation performed by the insect cells is not crucial for the binding of the usually highly glycosylated Fc gamma RII to IgG. The dissociation constant of the sFc gamma RIIa:IgG-hFc complex was determined by fluorescence titration (KD = 2.5 x 10(-)7 M). Complementary sFc gamma RIIa antagonizes immunocomplex binding to B cells. Here sFc gamma RIIa showed a comparable dissociation constant (KD = 1.7 x 10(-)7 M) which was almost 10-fold lower than the constant for Fc gamma RIIb. The stoichiometry of the FcRIIa:IgG hFc complex was determined by equilibrium gel filtration and shows that IgG is able to bind alternatively one or two Fc gamma RII molecules in a noncooperative manner. Furthermore, in an ELISA-based assay the isotype specificity of various anti-Fc gamma RII monoclonal antibodies was measured as well as their ability to interfere with the IgG recognition through its receptors. To further investigate the molecular basis of the Fc gamma RII-ligand interaction, we crystallized Fc gamma RIIb. Trigonal crystals diffracted to 3 A and the structure solution is in progress. PMID- 10387094 TI - Chimeric and truncated forms of human complement protein C8 alpha reveal binding sites for C8 beta and C8 gamma within the membrane attack complex/perforin region. AB - Human C8 is one of five components of the membrane attack complex of complement. It is an oligomeric protein composed of three subunits (C8 alpha, C8 beta, and C8 gamma) that are derived from different genes. C8 alpha and C8 beta are homologous and both contain a pair of tandemly arranged N-terminal modules [thrombospondin type 1 (TSP1) + low-density lipoprotein receptor class A (LDLRA)], an extended middle segment referred to as the membrane attack complex/perforin region (MACPF), and a pair of C-terminal modules [epidermal growth factor (EGF) + TSP1]. During biosynthetic processing, C8 alpha and C8 gamma associate to form a disulfide-linked dimer (C8 alpha-gamma) that binds to C8 beta through a site located on C8 alpha. In this study, the location of binding sites for C8 beta and C8 gamma and the importance of the modules in these interactions were investigated by use of chimeric and truncated forms of C8 alpha in which module pairs were either exchanged for those in C8 beta or deleted. Results show that exchange or deletion of one or both pairs of modules does not abrogate the ability of C8 alpha to form a disulfide-linked dimer when coexpressed with C8 gamma in COS cells. Furthermore, each chimeric and truncated form of C8 alpha gamma retains the ability to bind C8 beta; however, only those containing the TSP1 + LDLRA modules from C8 alpha are hemolytically active. These results indicate that binding sites for C8 beta and C8 gamma reside within the MACPF region of C8 alpha and that interaction with either subunit is not dependent on the modules. They also suggest that the N-terminal modules in C8 alpha are important for C9 binding and/or expression of C8 activity. PMID- 10387095 TI - Molecular mechanism of VanHst, an alpha-ketoacid dehydrogenase required for glycopeptide antibiotic resistance from a glycopeptide producing organism. AB - The vancomycin resistance enzyme VanH is an alpha-ketoacid dehydrogenase that stereospecifically reduces pyruvate to D-lactate, which is required for the synthesis of the depsipeptide D-alanine-D-lactate. This compound then forms an integral part of the bacterial cell wall replacing the vancomycin target dipeptide D-alanine-D-alanine, thus the presence of VanH is essential for glycopeptide resistance. In this work, the VanH homologue from the glycopeptide antibiotic producing organism Streptomyces toyocaensis NRRL 15009, VanHst, has been overexpressed in Escherichia coli and purified, and its substrate specificity and mechanism were probed by steady-state kinetic methods and site directed mutagenesis. The enzyme is highly efficient at pyruvate reduction with kcat/Km = 1.3 x 10(5) M-1 s-1 and has a more restricted alpha-ketoacid substrate specificity than VanH from vancomycin resistant enterococci (VRE). Conversely, VanHst shows no preference between NADH and NADPH while VanH from VRE prefers NADPH. The kinetic mechanism for VanHst was determined using product and dead-end inhibitors to be ordered BiBi with NADH binding first followed by pyruvate and products leaving in the order D-lactate, NAD+. Site-directed mutagenesis indicated that Arg237 plays a role in pyruvate binding and catalysis and that His298 is a candidate for an active-site proton donor. Glu266, which has been suggested to modulate the pKa of the catalytic His in other D-lactate dehydrogenases, was found to fulfill a similar role in VanHst, lowering a pKa value of kcat/Km nearly 2 units. These results now provide the framework for additional structure and inhibitor design work on the VanH family of antibiotic resistance enzymes. PMID- 10387096 TI - ATP nucleotidylation of creatine kinase. AB - Creatine kinase (CK) will autoincorporate radiolabel from [gamma32P]ATP and has thus been reported to be autophosphorylated. Also, in contrast to normal brain enzyme, CK in Alzheimer-diseased brain homogenate shows greatly decreased activity, abolished photolabeling with [32P]8N3ATP, and no detectable autoincorporation of radiolabel by [gamma32P]ATP. Surprisingly, our studies with both human brain and purified CK showed that [alpha32P]ATP, [gamma32P]ATP, [alpha32P]ADP, [2,8H3]ATP, [gamma32P]2',3'-O-(2,4, 6-trinitrophenyl)-ATP, and [gamma32P]benzophenone-gammaATP all autoincorporate radiolabel into CK with good efficiency. This demonstrates that the gamma-phosphate and the 2' and 3' hydroxyls are not involved in the covalent linkage and that all three phosphates, the ribose and base of the ATP molecule are retained upon autoincorporation (nucleotidylation). Treatment with NaIO3 to break the 2'-3' linkage effected total loss of radiolabel indicating that nucleotidylation resulted in opening of the ribose ring at the C1' position. Nucleotidylation with increasing [alpha32P]ATP at 37 degrees C gives an approximate k0.5 of 125 microM and saturates at 340 microM nucleotide. Modification of 8-10% of the copy numbers occurs at saturation, and CK activity is inhibited to approximately the same degree. Low micromolar levels of native substrates such as ADP, ATP, and phosphocreatine substantially reduce [alpha32P]ATP nucleotidylation. In contrast, AMP, GTP, GMP, NADH, and creatine did not effectively reduce nucleotidylation. When [alpha32P]ATP-nucleotidylated or [alpha32P]8N3ATP-photolabeled CK is treated with trypsin a single, identical radiolabeled peptide (V279-R291) is generated that comigrates on reverse phase HPLC and Tris-tricine electrophoresis. Nucleotidylation into this peptide was prevented 86% by the presence of ATP. We conclude that CK is nucleotidylated within the active site by modification at the C1'position and that autophosphorylation of this enzyme does not occur. PMID- 10387097 TI - Reactions of dimethylsulfoxide reductase from Rhodobacter capsulatus with dimethyl sulfide and with dimethyl sulfoxide: complexities revealed by conventional and stopped-flow spectrophotometry. AB - Improved assays for the molybdenum enzyme dimethylsulfoxide reductase (DMSOR) with dimethyl sulfoxide (DMSO) and with dimethyl sulfide (DMS) as substrates are described. Maximum activity was observed at pH 6.5 and below and at 8.3, respectively. Rapid-scan stopped-flow spectrophotometry has been used to investigate the reduction of the enzyme by DMS to a species previously characterized by its UV-visible spectrum [McAlpine, A. S., McEwan, A. G., and Bailey, S. (1998) J. Mol. Biol. 275, 613-623], and its subsequent reoxidation by DMSO. Both these two-electron reactions were faster than enzyme turnover under steady-state conditions, indicating that one-electron reactions with artificial dyes were rate-limiting. Second-order rate constants for the two-electron reduction and reoxidation reactions at pH 5.5 were (1.9 +/- 0.1) x 10(5) and (4.3 +/- 0.3) x 10(2) M-1 s-1, respectively, while at pH 8.0, the catalytic step was rate-limiting (62 s-1). Kinetically, for the two-electron reactions, the enzyme is more effective in DMS oxidation than in DMSO reduction. Reduction of DMSOR by DMS was incomplete below approximately 1 mM DMS but complete at higher concentrations, implying that the enzyme's redox potential is slightly higher than that of the DMS-DMSO couple. In contrast, reoxidation of the DMS-reduced state by DMSO was always incomplete, regardless of the DMSO concentration. Evidence for the existence of a spectroscopically indistinguishable reduced state, which could not be reoxidized by DMSO, was obtained. Brief reaction (less than approximately 15 min) of DMS with DMSOR was fully reversible on removal of the DMS. However, in the presence of excess DMS, a further slow reaction occurred aerobically, but not anaerobically, to yield a stable enzyme form having a lambdamax at 660 mn. This state (DMSORmod) retained full activity in steady-state assays with DMSO, but was inactive toward DMS. It could however be reconverted to the original resting state by reduction with methyl viologen radical and reoxidation with DMSO. We suggest that in this enzyme form two of the dithiolene ligands of the molybdenum have dissociated and formed a disulfide. The implications of this new species are discussed in relation both to conflicting published information for DMSOR from X-ray crystallography and to previous spectroscopic data for its reduced forms. PMID- 10387098 TI - Cryoenzymic studies on an organized system: myofibrillar ATPases and shortening. AB - We have exploited cryoenzymology, first, to probe the product release steps of myofibrillar ATPase under relaxing conditions and, second, to define the conditions for studying the contractile process in slow motion. Cryoenzymology implies perturbation by temperature and by the antifreeze added to allow for work at subzero temperatures. Here, we studied myofibrillar shortening and ATPases by the rapid quench flow method over a wide temperature range (-15 to 30 degrees C) in two antifreezes, 40% ethylene glycol and 20% methanol. The choice of solvent and temperature was dictated by the purpose of the experiment. Ethylene glycol (40%) is suitable for investigating the kinetics of the products release steps which is difficult in water. In this cryosolvent, the myofibrillar ATPase is not activated by Ca2+ nor is there shortening, except under special conditions, i.e., Ca2+ plus strong rigor bridges [Stehle, R., Lionne, C., Travers, F., and Barman, T. (1998) J. Muscl. Res. Cell Motil. 19, 381-392]. By the use of the glycol, we show that at low Ca2+ the kinetics of the ADP release are much faster with myofibrils than with S1. On the other hand, the kinetics of the Pi release were very similar for the two materials. Therefore, we suggest that, upon Ca2+ activation, only the Pi release kinetics are accelerated. In 20% methanol, in the presence of Ca2+, myofibrils shortened at temperatures above -2 degrees C but not below. At a given temperature above -2 degrees C, both the shortening and ATPase rates were reduced by the methanol. The temperature dependences of the myofibrillar ATPases (+/-Ca2+) converged with a decrease in temperature: at 20 degrees C, Ca2+ activated 30-fold, but at -15 degrees C, only about 5-fold. We suggest that studies in methanol may open the way for an investigation of muscle contraction in slow motion and, further, to obtain thermodynamic information on the internal forces involved in the shortening process. PMID- 10387099 TI - AE2 anion exchanger polypeptide is a homooligomer in pig gastric membranes: a chemical cross-linking study. AB - Although considerable information is available on the oligomeric states of the AE1 (band 3) anion exchanger, little is known about the physiological state of the polypeptides encoded by the nonerythroid AE genes, AE2 and AE3. We have previously characterized the proteolytic susceptibility of native pig gastric AE2. In the course of studies in which pig gastric membranes were treated with the AE2 transport antagonist, DIDS, we noted evidence for cross-linking of AE2 proteolytic fragments to higher-order oligomeric forms. We have characterized the ability of DIDS and of selected N-hydroxysuccinimide cross-linking agents to increase the proportion of SDS-resistant oligomers of pig gastric AE2 and its proteolytic fragments. Cross-linking exhibited time and concentration dependence. N-Terminal protein sequencing proved that DIDS treatment created AE2 homodimers. Putative homotetramers were also observed. Protomers were cross-linked via residues within the C-terminal 40 kDa of AE2. Prior proteolytic cleavage of AE2 in membranes resulted in decreased yield of subsequently cross-linked products. AE2 cross-linking could not be detected in membranes pretreated by hypotonic wash and freeze-thaw. The results are interpreted in light of the deduced amino acid sequence of the transmembrane domain of pig AE2. PMID- 10387100 TI - Apocalmodulin and Ca2+ calmodulin bind to the same region on the skeletal muscle Ca2+ release channel. AB - The skeletal muscle Ca2+ release channel (RYR1) is regulated by calmodulin in both its Ca2+-free (apocalmodulin) and Ca2+-bound (Ca2+ calmodulin) states. Apocalmodulin is an activator of the channel, and Ca2+ calmodulin is an inhibitor of the channel. Both apocalmodulin and Ca2+ calmodulin binding sites on RYR1 are destroyed by a mild tryptic digestion of the sarcoplasmic reticulum membranes, but calmodulin (either form), bound to RYR1 prior to tryptic digestion, protects both the apocalmodulin and Ca2+ calmodulin sites from tryptic destruction. The protected sites are after arginines 3630 and 3637 on RYR1. These studies suggest that both Ca2+ calmodulin and apocalmodulin bind to the same or overlapping regions on RYR1 and block access of trypsin to sites at amino acids 3630 and 3637. This sequence is part of a predicted Ca2+ CaM binding site of amino acids 3614-3642 [Takeshima, H., et al. (1989) Nature 339, 439-445]. PMID- 10387101 TI - Quantitative assessment of mRNA cap analogues as inhibitors of in vitro translation. AB - Fifty-eight analogues of the 5'-terminal 7-methylguanosine-containing cap of eukaryotic messenger RNA were synthesized and tested for their ability to inhibit in vitro protein synthesis. A new algorithm was developed for extracting KI, the dissociation constant for the cap analogue.eIF4E complex, from protein synthesis data. The results indicated that addition of a methyl group to the N2 of guanine produced more inhibitory compounds, but addition of a second methyl group to N2 decreased the level of inhibition dramatically. Aryl substitution at N7 improved the efficacy of guanine nucleoside monophosphate analogues. Substitution of the aromatic ring at the para position with methyl or NO2 groups abolished this effect, but substitution with Cl or F enhanced it. By contrast, aryl substitution at N7 in nucleoside di- or triphosphate analogues produced only minor effects, both positive and negative. By far the strongest determinants of inhibitory activity for cap analogues were phosphate residues. The beneficial effect of more phosphate residues was related more to anionic charge than to the number of phosphate groups per se. The second nucleotide residue in analogues of the form m7GpppN affected inhibitory activity in the order G > C > U > A, but there was no effect of 2'-O-modification. Opening the first ribose ring of m7GpppG analogues dramatically decreased activity, but alterations at the 2'-position of this ribose had no effect. Non-nucleotide-based cap analogues containing benzimidazole derivatives were inhibitory, though less so than those containing 7 methylguanine. PMID- 10387102 TI - Specificity in the binding of aminoglycosides to HIV-RRE RNA. AB - Quantitative studies of the binding of neomycin B to RRE constructs are carried out to determine the relationship between non-Watson Crick base-paired elements in the RNA and aminoglycoside binding. The RRE region contains two unpaired domains containing a single base bulge and a bubble structure, respectively. Deletion of the single base bulge has no effect on neomycin binding as the site of aminoglycoside binding is localized to the bubble region. Converting the bubble region into an A-form duplex gradually abolishes neomycin B binding in 3-5 fold steps in affinity over a 75-fold range. Thus, the binding of aminoglycoside is favored at domains in RNA that are nonduplex in nature, but aminoglycoside binding is only graded-specific in that affinities are enhanced gradually as the structure further deviates from a duplex form. It is likely that high-affinity aminoglycoside binding does not occur in duplex RNA because the major groove is too narrow to allow for aminoglycoside access and that structural perturbations that allow widening of the groove facilitate access. However, these interactions are only graded-specific with respect to both aminoglycoside structure and RNA domain structure. PMID- 10387103 TI - A pre-steady-state kinetic analysis of substrate binding to human recombinant deoxycytidine kinase: a model for nucleoside kinase action. AB - Deoxycytidine kinase (dCK) is an enzyme with broad substrate specificity which can phosphorylate pyrimidine and purine deoxynucleosides, including important antiviral and cytostatic agents. In this study, stopped-flow experiments were used to monitor intrinsic fluorescence changes induced upon binding of various phosphate donors (ATP, UTP, and the nonhydrolyzable analogue AMP-PNP) and the acceptor dCyd to recombinant dCK. Monophasic kinetics were observed throughout. The nucleotides as well as dCyd bound to the enzyme by a two-step mechanism, involving a rapid initial equilibrium step, followed by a protein conformational change that is responsible for the fluorescence change. The bimolecular association rate constants for nucleotide binding [(4-10) x 10(3) M-1 s-1] were 2 3 orders of magnitude lower than those for dCyd binding [(1.3-1.5 x 10(6) M-1 s 1]. This difference most likely is due predominantly to the large difference in the forward rate constants of the conformational changes (0.04-0.26 s-1 vs 560 710 s-1). Whereas the kinetics of the binding of ATP, UTP, and AMP-PNP to dCK showed some differences, UTP exhibiting the tightest binding, no significant differences were observed for the binding of dCyd to dCK in the presence or absence of phosphate donors. However, the binding of dCyd to dCK in the presence of ATP or UTP was accompanied by a 1.5- or 3-fold higher quenching amplitude as compared with dCyd alone or in the presence of AMP-PNP. We conclude that ATP and UTP induce a conformational change in the enzyme, thereby enabling efficient phosphoryl transfer. PMID- 10387104 TI - Rapid folding of calcium-free subtilisin by a stabilized pro-domain mutant. AB - In vitro folding of mature subtilisin is extremely slow. The isolated pro-domain greatly accelerates in vitro folding of subtilisin in a bimolecular reaction whose product is a tight complex between folded subtilisin and folded pro-domain. In our studies of subtilisin, we are trying to answer two basic questions: why does subtilisin fold slowly without the pro-domain and what does the pro-domain do to accelerate the folding rate? To address these general questions, we are trying to characterize all the rate constants governing individual steps in the bimolecular folding reaction of pro-domain with subtilisin. Here, we report the results of a series of in vitro folding experiments using an engineered pro domain mutant which is independently stable (proR9) and two calcium-free subtilisin mutants. The bimolecular folding reaction of subtilisin and proR9 occurs in two steps: an initial binding of proR9 to unfolded subtilisin, followed by isomerization of the initial complex into the native complex. The central findings are as follows. First, the independently stable proR9 folds subtilisin much faster than the predominantly unfolded wild-type pro-domain. Second, at micromolar concentrations of proR9, the subtilisin folding reaction becomes limited by the rate at which prolines in the unfolded state can isomerize to their native conformation. The simpliest mechanism which closely describes the data includes two denatured forms of subtilisin, which form the initial complex with proR9 at the same rate but which isomerize to the fully folded complex at much different rates. In this model, 77% of the subtilisin isomerizes to the native form slowly and the remaining 23% isomerizes more rapidly (1.5 s-1). The slow-folding population may be unfolded subtilisin with the trans form of proline 168, which must isomerize to the cis form during refolding. Third, in the absence of proline isomerization, the rate of subtilisin folding is rapid and at [proR9] 3 s-1. The implications of these results concerning why subtilisin folds slowly without the pro-domain are discussed. PMID- 10387105 TI - Nature of oxygen activation in glucose oxidase from Aspergillus niger: the importance of electrostatic stabilization in superoxide formation. AB - Glucose oxidase catalyzes the oxidation of glucose by molecular dioxygen, forming gluconolactone and hydrogen peroxide. A series of probes have been applied to investigate the activation of dioxygen in the oxidative half-reaction, including pH dependence, viscosity effects, 18O isotope effects, and solvent isotope effects on the kinetic parameter Vmax/Km(O2). The pH profile of Vmax/Km(O2) exhibits a pKa of 7.9 +/- 0.1, with the protonated enzyme form more reactive by 2 orders of magnitude. The effect of viscosogen on Vmax/Km(O2) reveals the surprising fact that the faster reaction at low pH (1.6 x 10(6) M-1 s-1) is actually less diffusion-controlled than the slow reaction at high pH (1.4 x 10(4) M-1 s-1); dioxygen reduction is almost fully diffusion-controlled at pH 9.8, while the extent of diffusion control decreases to 88% at pH 9.0 and 32% at pH 5.0, suggesting a transition of the first irreversible step from dioxygen binding at high pH to a later step at low pH. The puzzle is resolved by 18O isotope effects. 18(Vmax/Km) has been determined to be 1.028 +/- 0.002 at pH 5.0 and 1.027 +/- 0.001 at pH 9.0, indicating that a significant O-O bond order decrease accompanies the steps from dioxygen binding up to the first irreversible step at either pH. The results at high pH lead to an unequivocal mechanism; the rate limiting step in Vmax/Km(O2) for the deprotonated enzyme is the first electron transfer from the reduced flavin to dioxygen, and this step accompanies binding of molecular dioxygen to the active site. In combination with the published structural data, a model is presented in which a protonated active site histidine at low pH accelerates the second-order rate constant for one electron transfer to dioxygen through electrostatic stabilization of the superoxide anion intermediate. Consistent with the proposed mechanisms for both high and low pH, solvent isotope effects indicate that proton transfer steps occur after the rate limiting step(s). Kinetic simulations show that the model that is presented, although apparently in conflict with previous models for glucose oxidase, is in good agreement with previously published kinetic data for glucose oxidase. A role for electrostatic stabilization of the superoxide anion intermediate, as a general catalytic strategy in dioxygen-utilizing enzymes, is discussed. PMID- 10387106 TI - Identification of the two zinc-bound cysteines in the ferric uptake regulation protein from Escherichia coli: chemical modification and mass spectrometry analysis. AB - Selective chemical modification of thiol groups combined with mass spectrometry analysis was used to characterize cysteine ligands in the zinc-binding site of the Fur protein. Fur is a metalloregulatory protein involved in the regulation of almost all bacterial genes related to iron uptake in Gram-negative bacteria such as Escherichia coli. In addition to the iron site, Fur also possesses a tight binding zinc site that likely comprises two cysteines. Using a new procedure, we confirm the involvement of two cysteines in zinc binding and identify them within the two pairs of cysteines present in the protein. The protein was treated under nondenaturing conditions with iodoacetamide, and the progressive alkylation of the thiol groups monitored by quenching the reaction at different times and measuring the extent of alkylation by mass spectrometry. Complementary experiments were carried out in the absence or presence of EDTA, a strong zinc chelator, to determine which of the cysteines were protected from alkylation by the zinc atom. Enzymatic digestion of the modified protein and analysis of the peptide mixture by mass spectrometry enabled fast identification of reactive and protected thiol groups. Two cysteines, Cys92 and Cys95, were thus assigned as zinc ligands. Examination of the sequence comprising the zinc site indicates that it may belong to a new type of structural zinc site. Furthermore, Cys132 was shown to be the fastest reacting cysteine, implying it is a surface-exposed residue. PMID- 10387108 TI - Temperature and pH dependence of the metarhodopsin I-metarhodopsin II equilibrium and the binding of metarhodopsin II to G protein in rod disk membranes PMID- 10387107 TI - Estimating loop-helix interfaces in a polytopic membrane protein by deletion analysis. AB - Insertions of amino acids into transmembrane helices of polytopic membrane proteins disrupt helix-helix interactions with loss of function, while insertions into loops have little effect on transmembrane helices and therefore little effect on activity [Braun, P., Persson, B., Kaback, H. R., and von Heijne, G. (1997) J. Biol. Chem. 272, 29566-29571]. Here the inverse approach, amino acid deletion, is utilized systematically to approximate loop-helix boundaries in the lactose permease of Escherichia coli. Starting with deletion mutants in the periplasmic loop between helices VII and VIII (loop VII/VIII), which has been defined by immunological analysis and nitroxide-scanning electron paramagnetic resonance spectroscopy, it is shown that mutants with single or multiple deletions in the central portion of the loop retain significant transport activity, while deletion of amino acid residues near the loop-helix boundaries or within the flanking helices leads to complete inactivation. Results consistent with hydropathy analysis are obtained with loops VI/VII, VIII/IX, and IX/X and the flanking helices. In contrast, deletion analysis of loops III/IV, IV/V, and V/VI and the flanking helices indicates that this region of the permease differs from hydropathy predictions. More specifically, evidence is presented supporting the contention that Glu126 and Arg144 which are charge paired and critical for substrate binding are within helices IV and V, respectively. PMID- 10387109 TI - Kinetic properties of purified Pseudomonas aeruginosa phosphorylcholine phosphatase indicated that this enzyme may be utilized by the bacteria to colonize in different environments. AB - Pseudomonas aeruginosa phosphorylcholine phosphatase from periplasmic extracts of bacteria grown on choline as the sole carbon and nitrogen source was purified to homogeneity. The enzyme represented nearly 1% of the total protein found in the periplasmic space and is a monomer of approximately 53 kDa with an isoelectric point of 7.5. The optimum pH with phosphorylcholine was in the range of 5-8; with phosphorylethanolamine there was a peak at pH 6, and with p-nitrophenyl-phosphate (p-NPP) the optimum was at pH 5. Studies carried out at pH 5 indicated: i) For the three substrates above, Mg2+, Zn2+, or Cu2+ was necessary for maximal activity. ii) With p-NPP, these cations bound to the free enzyme in an ordered bireactant system. iii) With phosphorylethanolamine, Mg2+, Zn2+, or Cu2+ bound to the free enzyme in an at random bireactant system. iv) With phosphorylcholine, maximal activity was obtained with cation concentrations as low as 100 nM. v) Al3+ ions were inhibitors of the enzyme activity. The n (Hill coefficient) values for the inhibition by Al3+ with phosphorylcholine or p-NPP were 1 or 4, respectively. vi) The enzyme exhibited two affinity sites for phosphorylcholine. With Mg2+, a site with a Km value of 0.5 mM was detected; the corresponding Vmax was 25 micromol min-1 (mg protein)-1; without Mg2+, the enzyme displayed Km and Vmax values of 0.09 mM and 4.2 micromol min-1 (mg protein)-1, respectively. Studies carried out at pH 7.4 indicated: i) The enzyme could not catalyze the hydrolysis of p-NPP, and phosphorylethanolamine was a poor substrate in either the presence or absence of divalent cations. ii) The enzyme activity measured with phosphorylcholine was independent of divalent cations or was not inhibited by Al3+ ions. iii) With or without Mg2+, the enzyme exhibited two affinity sites for phosphorylcholine; the Km values were 0.05 mM and 0.5 mM with their corresponding Vmax of 5.6 and 25 micromol min-1 (mg protein)-1, respectively. PMID- 10387111 TI - Binding of phylogenetically distant Bacillus thuringiensis cry toxins to a Bombyx mori aminopeptidase N suggests importance of Cry toxin's conserved structure in receptor binding. AB - We investigated the binding proteins for three Cry toxins, Cry1Aa, Cry1Ac, and the phylogenetically distant Cry9Da, in the midgut cell membrane of the silkworm. In a ligand blot experiment, Cry1Ac and Cry9Da bound to the same 120-kDa aminopeptidase N (APN) as Cry1Aa. A competition experiment with the ligand blot indicated that the three toxins share the same binding site on several proteins. The values of the dissociation constants of the three Cry toxins and 120-kDa APN are as low as the case of other Cry toxins and receptors. These results suggest that distantly related Cry toxins bind to the same site on the same proteins, especially with APN. We propose that the conserved structure in these three toxins includes the receptor-binding site. PMID- 10387110 TI - Expression of orf1 from the Bacillus thuringiensis NRD-12 cry2Aa1 operon. AB - The 5' untranslated region and the orf1 sequence from the cry2Aa1 operon from Bacillus thuringiensis subsp. kurstaki NRD-12 were sequenced and compared to that from strain HD-1. The start codon described in HD-1 does not yield in NRD-12 a protein of the expected size of 20 kDa, but a 10-amino acid peptide. A second, highly conserved start codon is located 25 bp downstream from the first one and corresponds to an open reading frame of the same size in all known orf1-related sequences. Expression of lacZ gene fusions created at the level of the first ATG, second ATG, and stop codon of the NRD-12 orf1 sequence showed that orf1 is translated from the second ATG. The expected protein is 19 kDa in size. The expression starts at t2, which is in agreement with the presence of a BtI promoter in the cry2Aa1 operon. PMID- 10387112 TI - Sanitation of wallboard colonized with Stachybotrys chartarum. AB - Sections (8 cm2) of unused, nonsterile gypsum wallboard (dry wall) were inoculated with varying densities (10(4) to approximately 10(8)/ml) of conidia from 14- to 21-day cultures of Stachybotrys chartarum grown on cellulose agar. The sections were permitted to air dry and were placed into vessels with 86% or 92% RH and incubated at 22-25 degrees C for up to 12 weeks. The moisture content of the dryboard increased from near 10% to over 35%. Selected sections with confluent surface growth, mainly of S. chartarum, were obtained within 3 weeks. Sections were cleaned with a quaternary or quaternary and chlorine dioxide or a concentrated oxygen-saline solution and treated, in some cases, with a preservative system and returned to humidity vessels. Reemergence of S. chartarum from inoculated and treated surfaces occurred within 5 weeks only with sections treated with the quaternary alone. Other fungi, mostly species of Aspergillus, Chaetomium and Penicillium, slowly colonized (between 9-12 weeks) at least some areas of most treated surfaces and most uninoculated control surfaces. Stachybotrys chartarum was also found on several sections of uninoculated controls. Sections treated with a quaternary/acrylic and placed in a dynamic challenging chamber remained visually free of colonized fungi for over 90 days. These studies indicate that control samples of uninstalled wallboard, available from local distributors, can contain a baseline bioburden, including S. chartarum, that will colonize surfaces under high humidity conditions. Sanitation and preservation treatment of the wallboard can markedly delay regrowth of these fungi, particularly of S. chartarum. PMID- 10387113 TI - Presence of Na+-stimulated P-type ATPase in the membrane of a facultatively anaerobic alkaliphile, Exiguobacterium aurantiacum. AB - It was found that a facultatively anaerobic alkaliphile, Exiguobacterium aurantiacum, possesses a membrane-bound ATPase, which was activated specifically by Na+. The Na+-stimulated ATPase activity reached a maximum value at 200 mM NaCl. In the presence of 200 mM NaCl, the activity was drastically reduced by vanadate, a potent inhibitor of P-type ATPase, with a half-maximal inhibition at 1 microM. Incubation of the membranes with [gamma-32P]ATP followed by acidic lithium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated the existence of two phosphorylated intermediates with apparent molecular masses of 60 and 100 kDa. Only phosphorylation of the 100-kDa polypeptide was inhibited by vanadate. The membrane extract containing Na+-stimulated ATPase, when reconstituted into soybean phospholipid vesicles, exhibited 22Na+ transport by the addition of ATP, which was inhibited by vanadate and gramicidin. It is likely that the Na+-stimulated ATPase belongs to P-type and is involved in Na+ transport. PMID- 10387114 TI - Cloning, sequencing, and chromosomal location of a putative class-II aldolase gene from Streptococcus pneumoniae. AB - The nucleotide sequence of a 1620-bp chromosomal fragment from Streptococcus pneumoniae, containing a putative class-II aldolase gene, has been determined. The N-terminal amino acid (aa) sequence of S. pneumoniae class-II aldolase protein allowed us to determine the initiation site for the putative aldolase gene, and a molecular weight of 31,274 Da was predicted for the protein, after removal of the N-terminal methionine. Northern hybridization and primer extension analysis showed a 1100-nucleotide transcript with a transcription start site located 43 or 42 bp upstream of the start codon. Southern hybridization studies indicated that the putative class-II aldolase gene was in the ApaI fragment 6, SmaI fragment 9, and SacII fragment 12 or 13 of the physical map of S. pneumoniae chromosome. Southern hybridization analysis and partial sequencing performed in another eight streptococcus species, belonging to six different phylogenetic groups, suggested that a class-II aldolase gene with a considerable DNA homology to that of the S. pneumoniae, could exist in these streptococcal species. PMID- 10387115 TI - The LHIalpha and LHIIalpha complexes in association with phospholipids are able to be inserted in heavy membranes of Rhodobacter capsulatus B10. AB - We show in this paper that a complex constituted by phospholipids and LHI and LHII alpha polypeptides was inserted in a heavy membrane fraction in a nonextractable form, indicating a transmembrane localization. The best accepting membranes originated from aerobically grown cells. Addition of ATP during the insertion inhibited this reaction 25 to 30% in heavy membranes isolated from aerobically grown cells (HMaer) and a higher inhibition (60 to 65%) was detected when using heavy membranes isolated from photosynthetically grown cells (HMpho). Purification by gel filtration of a crude Na2CO3 extract yielded three phosphate labeled fractions. Two of them contained protein and phospholipids in a stable association. However, only fractions containing phosphatidylethanolamine were shown to be reconstituted. The third radioactive fraction contained labeled ATP and protein, but no phospholipids and could not be reassociated to the heavy membranes of any origin. A model for the insertion of the LH polypeptides is presented in which the recently synthesized polypeptides are phosphorylated and become associated to anionic phospholipids. The interaction of this complex to the membrane spontaneously leads to stable insertion. PMID- 10387116 TI - Characterization, production, and purification of leucocin H, a two-peptide bacteriocin from Leuconostoc MF215B. AB - Leuconostoc MF215B was found to produce a two-peptide bacteriocin referred to as leucocin H. The two peptides were termed leucocin Halpha and leucocin Hbeta. When acting together, they inhibit, among others, Listeria monocytogenes, Bacillus cereus, and Clostridium perfringens. Production of leucocin H in growth medium takes place at temperatures down to 6 degrees C and at pH below 7. The highest activity of leucocin H in growth medium was demonstrated in the late exponential growth phase. The bacteriocin was purified by precipitation with ammonium sulfate, ion-exchange (SP Sepharose) and reverse phase chromatography. Upon purification, specific activity increased 10(5)-fold, and the final specific activity was 2 x 10(7) BU/OD280. Amino acid composition analyses of leucocin Halpha and leucocin Hbeta indicated that both peptides consisted of around 40 amino acid residues. Their N-termini were blocked for Edman degradation, and the methionin residues of leucocin Hbeta did not respond to Cyanogen Bromide (CNBr) cleavage. Absorbance at 280 nm indicated the presence of tryptophan residues and tryptophan-fracturing opened for partial sequencing by Edman degradation. From leucocin Halpha, the sequence of 20 amino acids was obtained; from leucocin Hbeta the sequence of 28 amino acid residues was obtained. No sequence homology to other known bacteriocins could be demonstrated. It also appeared that the two peptides themselves shared little or no sequence homology. The presence of soy oil did not affect the activity of leucocin H in agar. PMID- 10387117 TI - Sorption and desorption of cobalt by Oscillatoria anguistissima. AB - Oscillatoria anguistissima rapidly adsorbs appreciable amounts of cobalt from the aqueous solutions within 15 min of initial contact with the metal solution. O. anguistissima showed a high sequestration of cobalt at low equilibrium concentrations, and it followed the Freundlich model of adsorption. The adsorption is a strongly pH-dependent and temperature-independent phenomenon. The presence of Mg2+ and Ca2+ (100-200 ppm) resulted in decline in Co2+ adsorption capacity of Oscillatoria biomass. Sulphate and nitrate (0. 75-10 mM) drastically reduced the extent of Co2+ biosorption. The biosorption of cobalt is an ion exchange process as the Co2+ binding was accompanied by release of a large amounts of Mg2+ ions. Na2CO3 (1.0 mM) resulted in about 76% desorption of Co2+ from the loaded biomass. PMID- 10387119 TI - Management of Carotid Artery Stenosis: A Review. AB - Carotid endarterectomy clearly benefits high stroke-risk patients, but its value for asymptomatic patients is still being debated. If a high exposure is necessary for redo procedures or distal aneurysms, mandibular subluxation and styloidectomy may be required. Perioperative mortality and morbidity are acceptably low. Restenosis occurs in few patients.http://link.springer ny.com/link/service/journals/00547/bibs/8n3p139.html PMID- 10387118 TI - Biomineralization of carbonates by Marinococcus albus and Marinococcus halophilus isolated from the Salar de Atacama (Chile). AB - We studied the precipitation of carbonates in 17 strains of moderately halophilic, Gram-positive cocci belonging to two species: Marinococcus halophilus and Marinococcus albus, isolated from the Salar de Atacama (Chile). They were cultivated in solid and liquid laboratory media for 42 days at salt concentrations (wt/vol) of 3%, 7.5%, 15%, and 20%. The bioliths precipitated were studied by X-ray diffraction and scanning electron microscopy. M. halophilus formed crystals at each of the salt concentrations, with a maximum number of strains capable of precipitating carbonates at 7.5% and 15% salt concentrations. M. albus did not precipitate at 20% and showed a maximum at 7.5%. This behavior is similar to that of other gram-positive bacteria and differs from that found in gram-negative bacteria. The bioliths precipitated were spherical, generally isolated, with a size of 10-100 microm, varying with salinity. They were of magnesium calcite (CO3 Ca1-x Mgx) with Mg content increasing with increasing salinity and Mg/Ca molar ratio of the culture medium. These results demonstrate the active role played by M. halophilus and M. albus in the precipitation of carbonates. PMID- 10387120 TI - The Effect of FR167653 on Cerebral Ischemia-Reperfusion Injury After Retrograde Cerebral Perfusion in a Canine Model. AB - Retrograde cerebral perfusion (RCP) has recently been reported to be useful for the repair of aortic arch aneurysms. However, there is a possibility that RCP supplies a limited amount of blood to the brain [1] and ischemia-reperfusion injury may occur after RCP. FR167653 (FR) is characterized as a potent suppressant of interleukin-1beta and tumor necrosis factor-alpha. We investigated the role of FR in preventing cerebral ischemia-reperfusion injury after RCP in a canine model. A total of 12 mongrel dogs was divided into two groups: in the FR group (n = 6), FR167653 (1 mg/kg/hour) was continuously administered during the period of RCP and rewarming; in the control group (n = 6), a physiological saline solution was administered at the same dosage as the FR167653 during the same period. Following hypothermia (20 degrees C) using cardiopulmonary bypass and circulatory arrest, RCP was performed by infusing oxygenated blood via the bilateral internal maxillary veins for 60 minutes at a perfusion pressure of 25 mmHg. The cerebral blood flow (CBF), cerebral metabolic rate for glucose (CMRGlu) and oxygen (CMRO2), and excretion of carbon dioxide (ExCO2) were measured. These results were expressed as the percentage of change from baseline values established immediately after anesthesia. CBF was significantly (p < 0.05) higher in the FR group than in the control group at 40 (159 +/- 25% and 82 +/- 21%, respectively) and 60 minutes (177 +/- 30% and 83 +/- 14%, respectively) after RCP. The lactate/pyruvate ratio of blood returned from the brain tissues was significantly (p < 0.05) lower in the FR group than in the control group at 40 and 60 minutes after RCP. CMRGlu was significantly (p < 0.05) higher in the FR group than in the control group 60 minutes after RCP. There was no significant difference in CMRO2 and ExCO2 between the two groups. It is concluded that FR167653 appears to be effective in protecting the brain from ischemia reperfusion injury after RCP.http://link.springer ny.com/link/service/journals/00547/bibs/8n3p143.html PMID- 10387121 TI - A Comparison of Safety and Efficacy of Sublingual Captopril with Sublingual Nifedipine in Hypertensive Crisis. AB - Sublingual nifedipine is commonly used in hypertensive crisis, however, it may result in several adverse effects such as reflex tachycardia, headache, and flushing. Research is continuing to find a new drug that has the same efficiency and fewer side effects. Sublingual captopril, a new preparation of angiotensin converting enzyme inhibitor, lowers blood pressure. It is not known whether it is effective in these emergent clinical settings. Therefore we designed a randomized, double-blind study to compare the efficacy and safety of those two drugs in hypertensive crisis. Eighty patients (32 male and 48 female) with hypertensive crisis were included in the study; their mean age was 43.4 +/- 7.9 years. Nifedipine 10 mg was given sublingually to 34 and captopril 25 mg to 46 patients randomly. There was no difference between the two drugs with respect to their antihypertensive effect. Heart rate significantly dropped (p < 0.01 and p < 0.001) in the patients taking captopril, but no changes were observed in the patients taking nifedipine. Twenty-three of 34 patients taking nifedipine encountered adverse effects. Adverse effects were observed in only three patients taking captopril (p < 0.001). Sublingual captopril is as effective as and has less side effects than sublingual nifedipine. Because sublingual captopril has fewer side effects, it may be safer than nifedipine in the treatment of hypertensive crisis.http://link.springer ny.com/link/service/journals/00547/bibs/8n3p147.html PMID- 10387122 TI - Recurrent Celiac Artery Compression Syndrome. AB - The celiac artery compression syndrome (CACS) is an infrequently described clinical condition with poorly defined diagnostic criteria and an obscure pathophysiology. It is usually associated with an extrinsic compression upon the celiac axis near its takeoff from the aorta by fibrous diaphragmatic bands or sympathetic neural fibers. We present a patient with CACS who suffered a recurrence of her original abdominal complaints.http://link.springer ny.com/link/service/journals/00547/bibs/8n3p150.html PMID- 10387123 TI - Lowered Total Intracellular Magnesium Status in a Subgroup of Hypertensives. AB - A new method to determine total Mg++ content in lymphocytes was developed, offering advantages for routine measurements as compared to fluorescence methods. Intracellular total Mg++ measurements were performed in lymphocytes of 18 healthy subjects and 19 untreated essential hypertensive patients. Mg++ content was referred to lymphocytic and membrane protein, which was determined according to Bradford's method. Mg++ measurements were performed by atomic absorption spectroscopy using a Video 12 apparatus of Thermo Electron Instrumentation Laboratory, Andover, USA. The results show that in patients with essential hypertension total intralymphocytic Mg++ content is significantly lower (0.07 +/- 0.05 mmol/g lymphocytic protein, mean +/- s.d.) as compared to controls (0.11 +/- 0.04 mmol/g lymphocytic protein, mean +/- s.d., p < 0.05). Free intracellular Mg++ content was measured in lymphocytes by the fluorescent indicator mag-fura II, showing no significant differences in normotensives and hypertensives (0.30 +/- 0.16 versus 0.38 +/- 0.17 mmol/l). Additionally, in platelets free intracellular Mg++ concentrations were not found of significant difference in normotensives and hypertensives (0.52 +/- 0.23 versus 0.47 +/- 0.27 mmol/l) using mag-fura-II. In plasma Mg++ concentrations there was no significant difference in the normotensive and hypertensive group (0.92 +/- 0.07 versus 0.88 +/- 0.07 mmol/l). There was no correlation between plasma or free or total cellular magnesium concentrations in both groups. Furthermore this method seems also suitable for routine measurements of total intracellular Mg++ concentrations in even larger measurements like mag-fur-II. Lowered total intracellular Mg++ concentrations in a subgroup of primary hypertensives may contribute to the development of this disorder, perhaps due to different buffering systems.http://link.springer-ny.com/link/service/journals/00547/bibs/8n3p154.html PMID- 10387124 TI - Unilateral Agenesis of Internal Carotid Artery with Subarachnoid Hemorrhage: Report of Two Cases. AB - Internal carotid artery (ICA) is a rare anomaly of embryologic development. Digital subtraction angiography examination showed no visualization of the ICA on the right side in a 30-year-old male patient and on the left side in a 47-year old female patient. Computed tomography (CT) revealed the absence of the corresponding bony carotid canal. Doppler examinations of the common carotid and external carotid arteries on the affected sides demonstrated high-resistance flow characteristics. Two cases of ICA agenesis with subarachnoid hemorrhage were presented and the literature is reviewed.http://link.springer ny.com/link/service/journals/00547/bibs/8n3p157.html PMID- 10387125 TI - Asymptomatic Pericardial Hydatid Cyst. AB - Cardiac involvement is rare in hydatid disease, but it carries a significant risk of potentially lethal complications. Cardiac hydatid cysts are mostly intramyocardial. Pericardial hydatid cysts without myocardial involvement are much less frequent. Cardiac imaging techniques, particularly two-dimensional echocardiography, are more useful in the detection of cardiac cysts. We present an incidentally detected, asymptomatic, pericardial hydatid cyst.http://link.springer-ny.com/link/service/journals/00547/bibs/8n3p161.html PMID- 10387126 TI - Ventricular Arrhythmia Suppression by Magnesium Treatment after Coronary Artery Bypass Surgery. AB - Ventricular arrhythmias occur frequently shortly after coronary artery bypass grafting (CABG), and their occurrence coincides with the postoperative decline in serum magnesium (Mg) levels. To examine if this decline causes ventricular arrhythmias and if their appearance could be reduced by intravenous Mg administration, 140 consecutive CABG patients were randomized to receive 70 mmol of Mg sulphate (N = 69) or placebo (N = 71) over two days. Serum Mg concentration fell to 0.77 mmol/l in the control group but rose to 1.09 mmol/l in the Mg group (p < 0.001). On 48 h Holter, the number of ventricular premature complexes (VPC) on the third postoperative day was reduced in the Mg group (4 +/- 5 vs 12 +/- 21 VPCs/h; p < 0.05) and the incidence of complex ventricular arrhythmias (Lown grade 2-5) was significantly diminished (19% vs 41% of the patients; p < 0.05). In multivariate analysis, high risk ventricular arrhythmias (repetitive polymorphic ventricular complexes, couplets, R-on-T complexes or operative tachycardia) were independently predicted by high number of bypassed vessels (p = 0.01), poor NYHA functional class (p = 0.06), preoperative diuretic use (p = 0.07), and low postoperative Mg levels (p = 0.08). In conclusion, correction of the postoperative decline in serum Mg concentration decreases the occurrence of early VPCs and complex ventricular arrhythmias. Patients with extensive underlying coronary artery disease and prior diuretic therapy appear to benefit greatest from Mg treatment.http://link.springer ny.com/link/service/journals/00547/bibs/8n3p165.html PMID- 10387127 TI - End-Stage Renal Disease and Coronary Artery Bypass Grafting. PMID- 10387129 TI - 18(th) Annual CongressThe Phlebology Society of AmericaDenver, Colorado * April 18-21, 1999Abstracts and Posters Presented. PMID- 10387130 TI - Erratum. PMID- 10387128 TI - Re: Implications of End-Stage Renal Disease on Cardiac Surgery. PMID- 10387131 TI - Electrophysiologic study in patients with atrial fibrillation: an idea whose time has come yet again. PMID- 10387132 TI - Radiofrequency catheter ablation of common atrial flutter: role of the eustachian valve. AB - INTRODUCTION: During radiofrequency catheter ablation of a common atrial flutter between the tricuspid annulus and the Eustachian valve "septal isthmus", double potentials were recorded along the Eustachian valve, previously described as an anatomical line of conduction block between the coronary sinus ostium and the inferior vena cava. RESULTS: Just before flutter termination, lengthening and beat to beat delay variations between the 2 components of the double potentials were correlated with simultaneous modifications of the flutter cycle length. CONCLUSION: The "septal isthmus" is a common pathway for the flutter wavefront and the impulse generating the second component of the double potential. It is also a good target for flutter ablation. PMID- 10387133 TI - Safety and efficacy of outpatient transseptal radiofrequency ablation of atrioventricular accessory pathways. AB - A retrospective analysis of 60 consecutive patients who underwent outpatient transseptal radiofrequency ablation of left sided accessory pathways at Westchester County Medical Center/New York Medical College from September 1994 to December 1997 was performed. Patients were followed for a mean duration of 22 months. No complications either local or related to the transseptal method were observed. All patients had successful ablation of the accessory pathway. One patient had a recurrence of symptoms. This study suggests transseptal radiofrequency ablation of the left sided accessory pathways to be safe, feasible and an effective procedure when performed in an outpatient setting. These results were obtained at a high volume center with experience using the transseptal technique. PMID- 10387134 TI - Outpatient transseptal radiofrequency ablation of atrioventricular accessory pathways-ready for prime time? PMID- 10387135 TI - Low incidence of significant valvar insufficiency following retrograde aortic radiofrequency catheter ablation in young patients. AB - The incidence of significant valvar insufficiency at late (<6 month) follow-up was retrospectively evaluated in 27 young patients (age 4. 0-18.0 years) undergoing 29 ablation procedures via the retrograde aortic approach for left sided accessory connections in whom pre-ablation and post-ablation echocardiograms were available for review. Valvar insufficiency was graded using color flow techniques as absent, trivial, mild, moderate, or severe by blinded reviewers. Ablation was acutely successful via the retrograde approach in 25 of 29 procedures among these 27 patients. Successful ablation was ultimately achieved in all 27 patients. At baseline, 7 patients had evidence of trivial or mild mitral insufficiency, and no patient had aortic insufficiency. Three patients had evidence of impaired left ventricular systolic performance in the presence of manifest pre-excitation. At follow-up, pre-existing mitral insufficiency resolved in 5/7 patients, and persisted in 2 patients. New mitral insufficiency was evident in 3 patients, and new aortic insufficiency was transiently evident in 1 patient following ablation (all trivial). Institutional experience (mean rank 10 cases vs. 33 cases, p <.0005), and lower patient weight (29.7 vs. 56.3 kilograms, p =.01) were the only factors associated with the development of new valvar insufficiency. Valvar insufficiency could not be detected by careful auscultation in any patient and was deemed clinically insignificant in all patients. We conclude that ablation of left-sided accessory connections can be performed via the retrograde aortic approach without creating clinically significant valvar insufficiency. PMID- 10387136 TI - Right posterior atrioventricular ring: a location for different types of atrioventricular accessory connections. AB - We present an unusual case of a 28-year-old female patient with recurrent episodes of tachycardias due to participation of two accessory connections located in the posterior tricuspid annulus. Both connections were of the atrioventricular type, the one with non decremental fast conducting properties at the right posteroseptal area, the other with node-like properties at the posterolateral tricuspid ring. Both pathways were successfully ablated transvenously with radiofrequency energy application at the same session. Implications about a common embryological origin of the two pathways as well as review of the literature for similar cases are presented. PMID- 10387138 TI - Unipolar recording in cardiac electrophysiologic studies. PMID- 10387139 TI - The effect of intravenous procainamide on the HV interval at electrophysiologic study. AB - The His bundle electrogram recorded at electrophysiologic study clearly differentiates atrioventricular (AV) node disease from distal conduction system disease. The distal conduction system may be tested further by infusing procainamide (10-15 mg/kg) intravenously. High-grade distal AV block or prolongation of the HV interval <80 ms was defined as an abnormal response to this test. We retrospectively reviewed the medical records of 79 patients who underwent electrophysiologic study with intravenous procainamide. An abnormal response to procainamide was observed in only 3% of 37 patients with a normal baseline HV ( or = 1.45%) or 6 (upper quartile, > or = 10%) months. However, MAWT scores were related only to expected waiting time (r = 0.47; P < 0.0001). CONCLUSIONS: Most patients reject waiting 6 months for elective CABG, even if offered along with a halving in surgical mortality (from 2% to 1%). Intolerance for further delay seems to be determined primarily by patients' attachment to their scheduled surgical dates. Many also have severely inflated perceptions of their risk of myocardial infarction in the queue. These results suggest a need for interventions to modify patients' inaccurate risk perceptions, particularly if a scheduled surgical date must be deferred. PMID- 10387410 TI - Open heart surgery in public and private practice. AB - OBJECTIVES: To compare open heart surgery services provided by public and private hospitals in Catalonia (Spain) according to case mix, procedures undergone and surgical mortality. METHODS: Data on all adult patients undergoing open heart surgery procedures were collected prospectively in a sample of public and privately owned centres for a 6.5-month period in 1994. Sociodemographic, clinical and procedural variables were collected. A predictive model stratifying patients according to their surgical mortality risk was used to adjust for differences in case mix between providers. RESULTS: Included were 1287 open heart surgery procedures. Public and private patients differed significantly in terms of gender, clinical history (e.g. hypertension, pulmonary disease, recent infarction) and procedural variables (e.g. reoperation, type of intervention). There were also statistically significant differences related to educational level, with better educated patients more likely to be treated in private centres. Crude surgical mortality rates differed between providers, although public centres operated on higher-risk patients. After adjusting for differences in case mix, the association between the type of provider and surgical mortality was not statistically significant (odds ratio 1.68; 95% CI from 0.94 to 3.0). CONCLUSIONS: Although crude mortality rates differ between public and private providers, there is a significant trend towards higher surgical risk in public centres. After adjusting for surgical risk, differences between types of provider decreased and were no longer statistically significant. The importance of other social and health-related factors, such as educational level, may explain differences between providers in their patients' surgical risk and in their performance in open heart surgery. PMID- 10387411 TI - Government funding of the UK National Health Service: what does the historical record reveal? AB - OBJECTIVES: To examine the historic funding record of the UK National Health Service (NHS) by year (1948-1997), political administration and political party. METHODS: Construction of a deflated expenditure series between 1948 and 1997 from published sources of cash spending for each UK country and by four main NHS budget heads using extrapolated NHS-specific inflation measures for each budget head. Analysis of the resultant real funding record for the UK NHS by year, political administration, political party and pre-general election years. RESULTS: A historical funding record constructed from a number of official sources appears to show noticeable differences in volume levels of government spending on the NHS in the UK between political administrations and between political parties. All administrations (apart from the 1951-1955 Churchill/Eden government) have increased funds to the NHS over and above the level of NHS specific inflation during their periods of office. Labour administrations have increased average annual real percentage funding by around 3.75% compared with an average increase of 2.33% for Conservative administrations. It does not appear that incumbent governments spend more than the long-run trend in pre-election (or, indeed, election) years. The economic difficulties of the mid 1970s (primarily the oil price shocks) appear to have realigned NHS spending at a lower level compared with spending rates in the 1950s, 1960s and early 1970s. Between 1950 and 1997, NHS cash spending as a percentage of gross domestic product increased by around 0.06% per year, with decreases in this proportion in 25 out of the 48 years examined. A comparatively crude analysis of changes in productive efficiency in the hospital and community health services sector between 1951 and 1991 suggests that there is no significant relationship between financial inputs (adjusted for NHS-specific inflation) and outputs (discharges and deaths). One explanation is that the NHS copes (at unknown cost) in times of financial stringency, but, conversely, does not systematically respond (at least, not in terms of increased output) in times of financial plenty. In policy terms, a very tentative interpretation of these findings would be that improvements in productivity can be brought about by restricting financial inputs and at the same time applying managerial pressure to the NHS to at least maintain (or improve) output levels (through, for example, improvements in medical practice). CONCLUSIONS: As a guide to voting, this analysis may confirm some prejudices. However, judging the performance of political administrations in relation to the NHS is rather more complex than a macro analysis of financial inputs alone suggests. The apparently weak relationship between inputs and outputs and the possible ability of governments to increase productivity by restricting inputs (and hence partially to deflect criticism of their funding policy) perhaps confirms other prejudices about the productive slack of large organisations. Again, however, care should be taken in the interpretation of the macro analysis, since the potential costs (e.g. reduced quality) arising from parsimonious funding are not captured by the global output measure used in this analysis. PMID- 10387412 TI - The influence of health needs assessment on health care decision-making in London health authorities. AB - OBJECTIVES: Health needs assessment gained prominence under the model of health care purchasing developed to support the 1991 reforms of the UK National Health Service (NHS). The objectives of this paper are to determine how needs assessment has been used in the NHS, to assess the influence it has had on decision-making, and to relate the observed uses of needs assessment to competing theoretical models of health care policy-making. METHODS: A survey of needs assessment activity in 14 London health authorities identified 217 needs assessments conducted between 1993 and 1996. Semi-structured interviews were conducted with public health and commissioning staff in each authority. RESULTS: The survey indicated that needs assessment directly supported decision-making and action in two-thirds of the studies identified, but up to 20% of needs assessments had no impact on service provision. Four key functions of health needs assessment were observed: identifying a problem; planning detailed changes to services; providing post hoc justification for earlier decisions; and using participation in needs assessment to build 'ownership' of subsequent decisions. CONCLUSIONS: The survey suggests that needs assessment is, in practice, consistent with a 'mixed scanning' model of decision-making. Needs assessment is used to help select issues for detailed investigation and to direct analytical and decision-making resources. However, certain key areas are not amenable to technical analysis and solution, and are resolved through bargaining. PMID- 10387413 TI - Are women more ready to consult than men? Gender differences in family practitioner consultation for common chronic conditions. AB - BACKGROUND: When consultations for all reasons are combined, women are seen to consult their general practitioners more than men through most of adult life. It is, therefore, often assumed that women are more likely to consult for every condition. OBJECTIVES: To examine whether women report being more likely to consult a general practitioner than men when taking account of the underlying condition and various aspects of the experience of the condition consulted for. METHODS: Home-based nurse-interviews with 852 people in early middle age (39 years) and 858 in late middle age (58 years) sampled from the general population in the West of Scotland. Detailed information about current chronic conditions included general practitioner consultation and reported experience of pain frequency, pain severity, limitation to normal activities and restricted activity in the previous four weeks. RESULTS: Women were no more likely than men to consult a general practitioner in the previous year when experiencing the five most common groups of conditions; in addition, women were no more likely than men to consult at a given level of severity for a given condition type, except in the case of one aspect of reported experience of mental health problems. CONCLUSIONS: The results argue against the most widely accepted explanation for gender differences in consulting, namely, that women are simply more likely to consult a general practitioner than men irrespective of underlying morbidity. Reasons for the higher rates of women consulting observed in general practice-based studies are discussed in relation to these data. PMID- 10387414 TI - Data-hungry physicians drive unique outcomes program. PMID- 10387415 TI - Health system taps decision support tools to plan new facility, new oncology unit. AB - Decision support "consultant on a disk." Health care executives are now using decision support tools to analyze community discharge data, competitor performance, and other key performance measures. Read about a New York system that used computer-generated reports to justify ambulatory care facilities and a formal oncology program. PMID- 10387416 TI - Physician urges providers to use report cards to compare performance of health plans. AB - Turning the tables on managed care organizations. Health systems are devising increasingly sophisticated report cards for physicians that examine a range of indicators. But when it comes to comparing health systems and plans, physicians are practically in the dark. One prominent managed care physician proposes the creation of a standardized report card for providers to rate managed care organizations. PMID- 10387417 TI - Successful medical groups set benchmarking standards for performance, practices. AB - Data Library: providers in the nation's most successful medical groups perform more procedures, use their physical space more efficiently, and have more support staff than those in peer organizations, according to these study results recently released by the Medical Group Management Association. PMID- 10387418 TI - HCIA study concludes top hospitals more likely to use low-cost first-line antibiotics. AB - When it comes to presurgical antibiotic prophylaxis, less is more. A benchmarking survey of America's top health systems shows that better-performing hospitals use fewer antibiotics, with no untoward effects on patients. Review the survey results. PMID- 10387419 TI - Report describes benchmarks for administrative expenses. AB - How does your HMO stack up against its peers where administrative costs are concerned? Data from a survey conducted by Sherlock Company of six large HMOs was broken out into six product lines and 15 major functions, and then analyzed three ways. Check out the results. PMID- 10387420 TI - HCFA outcomes improvement program for home health agencies raises privacy concerns. PMID- 10387421 TI - Beneficiary knowledge of the Medicare program. PMID- 10387422 TI - Changes to Medicare home health (as contained in the Omnibus Consolidated and Emergency Supplemental Appropriations Act for Fiscal Year 1999, Public Law 105 277). PMID- 10387423 TI - Choice of health plan: implications for access and satisfaction. AB - In this article, the authors examine why low-income persons choose a managed care plan and the effects of choice on access and satisfaction, using data from the 1995-96 Kaiser/Commonwealth Five-State Low-Income Survey. Two-thirds of those choosing a managed care plan cited costs or benefits as their primary reason. Logistic regressions indicate that choice of plan had a neutral or positive effect on access and satisfaction. Medicaid enrollees with choice were less likely than those without to have difficulty obtaining particular services, more likely to rate plan quality highly, and less likely to report major problems with plan rules. PMID- 10387424 TI - Urban health care in transition: challenges facing Los Angeles County. AB - The authors examine the Medicaid Section 1115 Demonstration Project currently underway in Los Angeles County. The waiver was designed as part of a response to a financial crisis the Los Angeles County Department of Health Services (LACDHS) faced in 1995. It provides financial relief to give the county time to restructure its system for serving the medically indigent population. Los Angeles County's goal is to reduce its traditional emphasis on emergency room and hospital care by building an integrated system of community-based primary, specialty, and public health care. This case study describes activities completed through the spring of 1997, approximately 1 year after the waiver was approved. PMID- 10387425 TI - Impact of report cards on employees: a natural experiment. AB - To determine the effect of survey-based, health plan report cards on employees as they selected their 1995 health plan, the authors surveyed two groups of Minnesota State employees, one of which received the report card and one that did not. Both groups were surveyed before and after their enrollment. The authors looked for report card effects on relative changes in the employees' knowledge of health plan benefits and their ratings of quality and cost attributes, as well as their plan choice, rates of switching plans, and willingness to pay higher premiums. The only report card effect found was an increase in perceived knowledge for employees with single coverage. PMID- 10387426 TI - Managed care's impact on Medicaid financing for early intervention services. AB - Medicaid has been a major source of financing for early intervention services since the inception of the Infants and Toddlers with Disabilities Program in 1986. In this article, the authors analyze Medicaid financing of early intervention services in 39 States before and after the introduction of managed care. The association between level of Medicaid financing and program characteristics, provider arrangements, managed care carve-out policies, and managed care contract requirements is assessed. The authors discuss the reduction of Medicaid financing after managed care and its implications for State Infants and Toddlers with Disabilities Programs, State Medicaid agencies, and managed care organizations. PMID- 10387427 TI - Serving rural Medicare risk enrollees: HMOs' decisions, experiences, and future plans. AB - This article identifies factors that influence health maintenance organizations' (HMOs) decisions about offering a Medicare risk product in rural areas; describes HMOs' recent experiences serving rural Medicare risk enrollees; and assesses the potential impact of Medicare program changes on the future willingness of HMOs to offer a Medicare risk product in rural areas. Data for the analysis were collected through interviews with a national sample of 27 HMOs. The results underscore the importance of adjusted average per capita cost (AAPCC) rates in HMOs' decisions to offer Medicare risk products in rural areas, but also indicate that other factors influence these decisions. PMID- 10387429 TI - NHS values. I want to tell you a story. AB - Many NHS staff feel that they are unable to put their values into action because of the constraints on the service. They want a working environment where trust is possible and mistakes can be made without retribution. The reality is often bullying and attempts at innovation being met with reproach. PMID- 10387428 TI - National health expenditures, 1997. AB - In 1997 health spending in the United States increased just 4.8 percent to $1.1 trillion. As a share of gross domestic product (GDP), national health expenditures (NHE) absorbed 13.5 percent of the country's output in 1997--a share that has remained relatively constant for 5 years. Despite the relative stability in recent years, signs of changing trends are emerging. PMID- 10387430 TI - Joint working. Out of step. PMID- 10387431 TI - Information technology. Looping the loop. AB - The demands of clinical governance will require major re-orientation of information systems in some hospitals. It is important that information systems provide clinical staff with access to relevant data from many sources. There are major issues of accuracy and security to be addressed in order to provide clinicians with the information they need. PMID- 10387432 TI - Drug management. Making introductions. PMID- 10387433 TI - Getting into the swing. PMID- 10387434 TI - On the evidence. Managing change. PMID- 10387435 TI - The current state of risk adjustment technology for capitation. AB - Risk adjustment for the purposes of making capitated payments better reflect the expected costs of medical care is a technology that is now being applied in the public and private sector. This article reviews the characteristics of many of the risk adjuster methods that have been put forward in recent years. Included are models based on diagnoses from inpatient hospital data, and from inpatient and ambulatory data. Models for the general population, Medicaid, and Medicare are discussed. Caveats in comparing models are also presented. PMID- 10387436 TI - The development of a risk-adjusted capitation payment system: the Maryland Medicaid model. AB - This article describes the risk-adjusted payment methodology employed by the Maryland Medicaid program to pay managed care organizations. It also presents an empirical simulation analysis using claims data from 230,000 Maryland Medicaid recipients. This simulation suggests that the new payment model will help adjust for adverse or favorable selection. The article is intended for a wide audience, including state and national policy makers concerned with the design of managed care Medicaid programs and actuaries, analysts, and researchers involved in the design and implementation of risk-adjusted capitation payment systems. PMID- 10387437 TI - Development of brief pictorial instruments for assessing spirituality in primary care. AB - The relationship between spirituality and health is an emerging area of study. However, spirituality assessment instruments with clinical utility for busy health care practices are lacking. The article describes research directed at developing and validating brief pictorial measures of spirituality as instruments to measure the relationship between reported spirituality and health. The instruments presented are patterned after the Dartmouth Medical School Primary Care Cooperative (COOP) charts. The charts also provide a nonthreatening way to enhance communication between patients and physicians in this important area. PMID- 10387438 TI - The incremental cost of second and third ambulatory procedures. AB - The discount factor applied to payments for second and subsequent ambulatory procedures in a payment system based on ambulatory patient groups (APGs) is important in determining the financial incentives of the system and the adequacy of the payment rates. A 1995 empirical study of data from all ambulatory surgery cases done in acute general hospitals in Maryland suggests that the incremental charges associated with an APG when it is a second procedure APG are about 24% of the charges that are incurred when the APG is the only procedure APG in an encounter. For the third procedure APG, the percentage drops to 16%. This 24% factor is much lower than the discount factor generally used in APG payment systems. The article presents APG-adjusted charges by payer to show differences in resource use by payer. These results will be useful for organizations developing APG-based payment systems for ambulatory surgery or desiring to use APGs for benchmarking purposes. PMID- 10387439 TI - Physician profiling using outpatient pharmacy data as a source for case mix measurement and risk adjustment. AB - Pharmacy data are a potential source of information on the health risks of members in a managed care organization (MCO). Reasons why these data can serve as a basis for case mix measurement of MCO populations are reviewed. Several pharmacy-based case mix measurement systems (CDS, KPARx) are summarized. Applications of pharmacy-based case mix systems for profiling of primary physician panels in an MCO are discussed and illustrated with several examples. PMID- 10387440 TI - Seeking asylum within ambulatory care. PMID- 10387441 TI - Urgent care and the emergency department: providing the right ambulatory care settings. AB - Emergency services are asked to treat a significant volume of patients that require nonemergent care. Alternative strategies have been proposed for managing this patient population, ranging from free-standing urgent care centers to the integration of urgent care patients with other patients treated in the ED. This article explores issues and physical options surrounding the provision of patient care of urgent care patients in the emergency services. PMID- 10387442 TI - New markets for ambulatory care: wellness center + physicians' office building + urgent care/diagnostic center. AB - As the shift to ambulatory care increases, many established acute care facilities are searching for ways to serve their communities more effectively, to maintain their position in the market--and to ensure continuing, healthy revenue streams. This article explores some of the issues faced by such an institution in a rapidly growing market in the Southeast. For this article, we refer to it as Metropolitan Hospital. PMID- 10387443 TI - Creating a new environment of ambulatory care: community health centers and the Planetree philosophy. AB - This article provides an overview of the approach, programming, and methodology for the renovation of the Peekskill Area Health Center, an urban based non-profit community health center in the Hudson Valley. The impetus of the renovation is in response to changes in the health care environment and overcrowding of the existing facility. The model development, which is reflected in the physical renovation, employees key elements of the Planetree approach to patient-centered care. Consistent with the national Community Health Center philosophy, the project is dedicated to vitalizing an economically depressed area through construction opportunities and links with local commercial businesses. PMID- 10387444 TI - New approaches to ambulatory care facilities in the United Kingdom--an investor developer's perspective. AB - The purpose of this article is to describe the strategic context within which ambulatory care facilities are being developed, to consider a range of models of care facilities available, and, by drawing on the experience of the authors, to comment on some of the investment and development issues arising from two projects in progress--the community hospitals in Richmond, Yorkshire, in rural north England, and Thames Ditton on the borders of outer south London. In the final section, we consider the possible future of ambulatory care development in the light of government policy. PMID- 10387445 TI - Small rural ambulatory care facilities in Middle America--Titusville Area Hospital, Titusville, Pennsylvania. AB - This case study is focused on Titusville Area Hospital because of current/recent activities at the hospital, but it also reflects previous experience with other facilities. The rural midwestern hospital no longer needs to be a single facility located in the largest community in the area. In fact, it can no longer be that single facility; it must service its population(s) at locations convenient to patients with the services appropriate to them. It is important that the hospital not only recognize its outreach areas through a satellite facility but also build a strong outpatient and support base at the hospital. PMID- 10387446 TI - Ambulatory care transitioning for the rural hospital. AB - As rural community hospitals continue their transition of health care delivery to predominantly outpatient and ambulatory services, many factors are being considered in design, planning, and operations to enact this change both efficiently and effectively. The following examines strategies that allow these changes to be incorporated successfully while maintaining the flexibility for the continued transition that these organizations will experience in the future. PMID- 10387447 TI - Innovation and customer focus drive successful ambulatory care programs. AB - The author identifies shortcomings in traditional hospital based ambulatory care programs. Characteristics of innovative, customer focused ambulatory care programs are delineated, and two examples in breast care and wound care are described. PMID- 10387448 TI - Organize for success: seven steps to a successful building project. AB - Intended for ambulatory care administrators considering a building project, the article discusses seven key steps to a successful project: Avoid common pitfalls: allow enough time, avoid over-delegation, focus staff on issues rather than egos, require staff "buy-ins" to project concepts and design, and understand the realties of construction. Select the right team: how to interview, assess qualifications, and make a sound decision. Understand all the project costs; construction cost is only one of many. Understand the various construction methods and pricing alternatives; become a sophisticated consumer. Understand the nature and scope of the risks inherent in building projects. Understand all the management issues. Start your project off right through producing. Identify strategic goals and needs, then quantify and qualify your project's intent, scope, and basic financial assumptions. PMID- 10387449 TI - Interior design for ambulatory care facilities: how to reduce stress and anxiety in patients and families. AB - The following article illustrates some important factors to consider when designing ambulatory care facilities (ACFs), and focuses on how wayfinding, noise control, privacy, security, color and lighting, general ambience, textures, and nature can have a profound influence on patient and family stress, consumer satisfaction, health and well-being. Other important design issues: convenience and accessibility, accommodation to various populations, consumer and family focus, patient education, image, as well as current equipment needs and future growth are examined in light of the prevailing trends in health care delivery. In sum, this feature explores the important stress-reducing and health-promoting elements involved in successful ACF design. PMID- 10387450 TI - Planning ahead: practical hints for designing ambulatory care facilities. AB - This article discusses a variety of topics that need to be addressed during the design of an ambulatory care facility, particularly those areas that need to be discussed at the beginning of the project. Topics include life safety/code issues involving building classification, employee safety, and accessibility; regulatory agencies; design issues including design sources, touring comparable facilities, room mock-ups, staff evaluation/wish list, setting standards, and determining responsibility; and medical equipment including planning and current trends. The time you spend planning ahead is time well spent. PMID- 10387451 TI - History of ambulatory care facilities from a roving hospital administrator's point of view. AB - Over the past four decades, the delivery of health care services has seen dramatic changes. Medicare coverage in 1966 expanded the ability of the health care industry to treat a growing segment of the population. Technology improvements have made great strides in the treatment of disease. Public policy decisions expanded health care coverage from an employee benefit to entitlement for large segments of the population. Managed care growth has reorganized the basic health care delivery system. The aggregate effect of all these items was a dramatic increase in the cost of providing health care in the United States. Beginning in the late 1960s, and escalating in the 1970s and 1980s, the increase of the cost of care was the primary issue for the industry. Only in the 1990s has the rate of increase modified through the impact of managed care, and now there is significant competition for the health care dollar among many competing providers. PMID- 10387452 TI - Health professionals and the international fight to ban land mines. PMID- 10387453 TI - PacifiCare rushes communications to its physicians, members and the community. AB - PacifiCare of Colorado educates key audiences on the company's physician contract negotiations and its desire to balance physician financial success with consumers' need for affordable health care premiums and to reassure members of its desire and goal to maintain the existing physician network. PacifiCare created two advertorials for the local newspaper to communicate its goal to the community members and physicians. PMID- 10387455 TI - 435 Project advocates more dollars for medical research. Pilot effort wins recognition from health marketers. Research!America, Alexandria, VA. PMID- 10387454 TI - Trident Health extends outreach in Charleston, S.C. Mall wellness center proves popular among lowcountry residents. PMID- 10387456 TI - "Grow up with me" slogan focuses on adult involvement with children. Kansas Health Foundation. PMID- 10387457 TI - "Control your diabetes" campaign encourages a healthy lifestyle. National Diabetes Education Program. PMID- 10387459 TI - America's Doctor Online provides easy access for consultations. AB - America's Doctor Online is offering a web site accessible to consumers, who want to engage in discussions with real physicians. In exchange, the company is partnering with hospitals and medical groups. If an Internet user needs a referral, the doctor will take the consumer's name and phone number and have a representative from the local-area sponsoring hospital call within 24 hours. PMID- 10387458 TI - Historic photos become hospital's PR, marketing tool. Washington Hospital, Fremont, CA. AB - In conjunction with a 50th anniversary, Washington Hospital in Fremont, Calif., unveiled a permanent exhibit of 145 photographs that chronicles the history of the Alameda County region east of San Francisco Bay. PMID- 10387460 TI - "Brooklyn attitude" grabs attention for hospital's ad campaign. Brooklyn Hospital Center, NY. PMID- 10387462 TI - Bibliography. Current world literature. Corneal and external disorders and refractive surgery. PMID- 10387461 TI - Losing the battle of the bugs. PMID- 10387463 TI - Management of postkeratoplasty astigmatism. AB - Many factors have led to the improved success rate for clear corneal grafts after penetrating keratoplasty. Unfortunately, postoperative corneal astigmatism commonly occurs and can produce significant visual impairment. Astigmatic correction may include spectacle correction or contact lenses, but if this fails, then surgical options are considered. Refractive surgical techniques such as suture removal or adjustment, relaxing incisions, wedge resections, and photorefractive keratectomy or laser in situ keratomileusis can dramatically reduce postoperative astigmatism after penetrating keratoplasty and lead to improved, functional vision. However, significant variability between results in individual patients can occur. So although general guidelines are useful, it is important to individualize and modify the planned surgery based on qualitative keratoscopy and corneal topography for the initial and subsequent astigmatic corrections. PMID- 10387464 TI - Informed consent in refractive surgery. AB - The informed consent process represents a key component in providing the highest quality of care for our patients and minimizing liability risk. Patients undergoing refractive surgery have high expectations for an excellent outcome and may become easily dissatisfied and upset if this is not achieved. Careful attention by the surgeon and supporting staff to educate the patient in a thorough and supportive fashion is paramount to this process. Breakdown in communication between physicians and patients combined with a poor outcome and feelings of mistrust often leads patients to consider malpractice litigation. The importance of understanding the key components and principles of the informed consent process cannot be over-emphasized and are presented in detail in this review. PMID- 10387465 TI - Indications, results, and complications of LASIK. AB - The technique of laser in situ keratomileusis (LASIK) has been used with very encouraging results in the treatment of all degrees of myopia and also shows considerable promise in the treatment of hyperopia. Compared with photorefractive keratectomy, LASIK is advantageous in causing minimal postoperative discomfort, in its rapid recovery of clear vision and stabilization of refractive change, in the infrequent occurrence of haze, and in its greater facility in correcting high degrees of myopia. However, LASIK is the more surgically demanding technique. We discuss our own experience with LASIK as well as published data from other centers. We anticipate that LASIK will continue to increase in importance in the surgical correction of refractive error. PMID- 10387466 TI - Antimetabolites in ocular surface neoplasia. AB - Malignant neoplasms of the ocular surface are uncommon but consist of squamous and melanocytic tumors. Surgical excision has been the mainstay of treatment and is perhaps the "gold standard." However, in those circumstances in which surgical intervention is not feasible, adjunctive treatments have been advocated. Recent attention has been given to the use of mitomycin C and 5-fluorouracil for the treatment of ocular surface squamous neoplasia. This article reviews the major publications relating to the use of antimetabolites in ocular surface neoplasia and highlights the most recent contributions to the ophthalmic literature. PMID- 10387467 TI - Topical nonsteroidal anti-inflammatory agents in ophthalmology. AB - Nonsteroidal anti-inflammatory agents or NSAIDs are potent inhibitors of prostaglandin synthesis. As such, they have found many useful roles in ophthalmology. NSAIDs are approved by the FDA to prevent intraoperative miosis during cataract surgery, reduce postoperative inflammation following cataract surgery, and control symptoms of allergic conjunctivitis and pain following refractive surgery. In addition, they have been shown to be effective in preventing cystoid macular edema following cataract surgery or treating cystoid macular edema once it occurs. PMID- 10387468 TI - Current diagnosis and treatment of corneal ulcers. AB - Successful treatment for a corneal ulcer requires proper diagnosis and antibiotic selection. The management should be guided by the severity of the clinical presentation, the ophthalmologist's confidence in making the proper diagnosis of bacterial keratitis, and the level of trust in the antibiotic agents chosen for the causative organisms. Universal standards include pretreatment cultures and dual broad-spectrum fortified antibiotics. Recent changes in practice suggest that empirical monotherapy treatment with a fluoroquinolone antibiotic may be appropriate for certain cases of bacterial keratitis. This article reviews the various diagnostic methods and treatment options currently practiced in the ophthalmic community. PMID- 10387469 TI - Advances in the diagnosis and management of keratoconjunctivitis sicca. AB - Keratoconjunctivitis sicca is a common ocular surface disease that develops in patients with aqueous tear deficiency. Recent advances have been made in diagnosis, pathogenesis, and therapy of this condition. Advances in diagnosis include improved understanding of the specificity of the tests used for diagnosis, elucidation of the mechanism of the ocular surface rose bengal and fluorescein staining that occurs in this condition, and the expanded use of impression cytology. Advances in pathogenesis include the concept that keratoconjunctivitis sicca is a condition of abnormal growth and differentiation and immune activation of the ocular surface epithelium. These findings indicate that keratoconjunctivitis sicca may represent a chronic wound-healing response to a poorly lubricated and inflamed ocular surface. Advances in therapy include improved nonpreserved artificial tears and therapies targeted at decreasing ocular surface inflammation. PMID- 10387470 TI - The current and future therapy of allergic conjunctivitis. AB - A wealth of antiallergic drugs is available for ocular allergy, and many new drugs will soon be approved. Pharmaceutical companies frequently seek approval of anti-inflammatory drugs for allergic indications, because it is relatively easy to perform clinical trials for ocular allergy. Extremely safe drugs for mild to moderate degrees of allergic conjunctivitis include antihistamines, mast cell stabilizers, and nonsteroidal anti-inflammatory agents. Topical corticosteroids, preferably those with reduced side effects, are available for more severe forms of ocular allergy. The choice of an antiallergic drug may be guided by the indication for which the drug was approved. The ultimate selection will be made based on the patient's symptoms, the drug's availability, and its cost. PMID- 10387471 TI - Clinical confocal microscopy. AB - Because it provides much higher magnification and better optical sectioning than a slit-lamp biomicroscope, confocal microscopy is ideally suited for clinical imaging of the cornea. One important clinical application of confocal microscopy has been the early detection and diagnosis of a number of infectious conditions, including infection with Acanthamoeba and microsporidium species, fungal keratitis, and contact lens-associated bacterial keratitis. Confocal microscopy has also been used for temporal evaluation of corneal wound healing following refractive surgery and penetrating keratoplasty. With the development of the new technique of quantitative confocal microscopy through-focusing, confocal microscopy can be used to measure epithelial, stromal, and corneal thickness accurately and reproducibly in human patients. Furthermore, conofocal microscopy through-focusing can be used to determine the initial photoablation depth, changes in epithelial, stromal, and corneal thickness, and subepithelial haze following photorefractive keratectomy. PMID- 10387472 TI - The complications of contact lens wear. AB - Out of the approximately 30 million contact lens wearers in North America, about 6%--approximately 1.8 million individuals--develop a complication each year [Acta Ophthalmol 1984, 62:556-565]. These problems range from self-limiting to sight threatening, and require rapid diagnosis and treatment to prevent vision loss. Contact lens complications are as varied as they are common, involving the lids, conjunctiva, and all layers of the cornea (epithelium, stroma, and endothelium). PMID- 10387473 TI - Intrastromal corneal ring technology: results and indications. AB - Intracorneal ring technology has shown rapid development in the past 10 years, and clinical results are confirming outstanding results for the correction of low to moderate refractive myopias. Two of these device designs, the Intrastromal Corneal Ring and the Intrastromal Corneal Ring Segments (both manufactured by KeraVision, Inc., Fremont, CA) are currently undergoing rigorous investigation in US Food and Drug Administration-regulated clinical trials. Results to date indicate the surgical procedure is safe and easily performed, the visual results are excellent (preserving best spectacle-corrected visual acuity), and the procedure provides stable and predictable correction through several years postoperatively. Preliminary results show that enhancements can be easily performed by device exchange, and the device can be removed, reversing the refractive effect. PMID- 10387474 TI - Detection, prevention, and rehabilitation of amblyopia. AB - Although there is a continuing effort to develop effective measures for detecting amblyopia, the debate continues in many countries as to the real value of population-wide pediatric vision screening programs. Nevertheless, amblyopia still represents a significant and treatable clinical problem for the practicing ophthalmologist. In this past year's literature, we can find that many visual functions are abnormal in amblyopia, but only a few are measured to reach a diagnosis or monitor treatment. Unfortunately, the wide variety of the measurement methods and the lack of definition for a cure of amblyopia are the cause of serious criticism of our "scientific" literature on the subject. All the while, old methods of treatment are resurfacing again: optical and pharmacological penalizations. The search for a possible "pill" to either replace or at least complement conventional methods of treatment also has continued in 1997 and 1998. As well, there have been quite a few papers on the long-term management of amblyopia. Finally, this review would not be complete without reporting on the "controversy du jour:" a report that challenges the need to cure amblyopia before operating on esotropia. PMID- 10387475 TI - Comitant strabismus. AB - Proprioceptive receptors have long been known anatomically to be present in extraocular muscles, specifically at the myotendinous junction. Their function in regulating smooth pursuits is experimentally demonstrated. The clinical significance of this for strabismus is still unknown. Esotropia surgery before resolution of moderate amblyopia is not detrimental. Botulinum toxin injections will correct infantile esotropia but require more anesthesia sessions overall than does conventional surgery, increasing cost. Late-onset comitant esotropia is usually refractive in nature and rarely neurologic. Comitant esodeviation is also prevalent in children with known neurologic insults. In both situations, it is the associated neurologic signs that point to the underlying neurologic cause. Posterior fixation suture will correct a high accommodative convergence/accommodation ratio esotropia. Exotropia negatively affects patients' quality of life. Surgical outcomes differ in patients whose angle of deviation increases with 1 hour of occlusion testing at extreme distances ("outdoor sensitivity"). Recession resection influences the distance and near angles of deviation equally whereas bilateral lateral rectus resection influences the distance deviation more than the near. Spray administration of cycloplegic agents to closed eyelids has been shown to be as effective as administration of eye drops. The spray is much better tolerated by patients and easier to administer. Photorefraction, although not yet effective as a screening tool, is useful to document alignment and refractive errors. PMID- 10387476 TI - Paralytic strabismus. AB - The use of sophisticated imaging techniques has led to a greater understanding of the mechanisms of some forms of strabismus. In particular, studies of strabismus in high myopia have demonstrated abnormal muscle courses and provided a basis for corrective surgery. Although injection of botulinum toxin has a place in modern strabismus management, there have been few controlled studies validating its use. An important paper in the review period directly compares botulinum with surgery to confirm its effectiveness for at least one indication. PMID- 10387477 TI - Neuro-ophthalmologic manifestations of systemic and neurologic disease. AB - Neuro-ophthalmology is a difficult and challenging field for both ophthalmologists and neurologists. Unfortunately, signs and symptoms of these patients can be very subtle and easily overlooked. Although neuro-ophthalmologic cases are rare in the practice of a general ophthalmologist, missing the diagnosis can have a severe impact on the patient's health. This paper reviews last year's publications on neuro-ophthalmologic manifestations of systemic and neurologic disease. PMID- 10387478 TI - Development of refraction and strabismus. AB - In the past year a number of studies have provided insights into the mechanisms whereby the eye can maintain coordinated growth to achieve emmetropia. Research into factors that may lead to a failure to emmetropize also has been promising. The effect of early spectacle intervention has been debated, with some evidence from animal studies suggesting that lenses may interfere with emmetropization. Human data on this topic are limited but do not appear to show the deleterious effects of lenses reported in the animal studies. The role of early astigmatism in the emmetropization process is not clear. Myopia research continues to hold promise for the eventual discovery of treatments to slow progression. With respect to the development of strabismus, there are many mechanisms for its induction. The problem is to identify the primary ones and their interactions. This article reviews some of the newer candidates, including pulleys that affect extraocular-muscle action and the role of nasally biased monocular optokinetic nystagmus. An understanding of the critical periods of the various visual dimensions involved in the development of strabismus is also crucial. PMID- 10387479 TI - Nystagmus. AB - This article reviews some of the past year's important papers, with emphasis on early onset and acquired neurological nystagmus. Advances in understanding the mechanisms of suppression of oscillopsia, the evolution of nystagmus, and the treatment of periodic alternating nystagmus and of nystagmus in albinism have been made in early-onset nystagmus. Successful pharmacological treatment for acquired neurological nystagmus has been demonstrated with the gamma aminobutyric acid agonist gabapentin and with memantine, a glutamate antagonist. PMID- 10387480 TI - Ocular and orbital trauma: preventive medicine in ophthalmology. PMID- 10387481 TI - Orbital infections and inflammations. AB - Orbital infections and inflammations present to the clinician with similar findings: periorbital edema, erythema, proptosis, and pain. History and clinical examination determine the work-up required to better define the disease process. Orbital infections continue to be associated primarily with diseases of the paranasal sinuses. Haemophilus influenza type B is no longer a significant pathogen, because of an effective vaccine. Fungal infections extending to the orbit are becoming more frequent due to the prevalence of immunocompromised patients. Orbital inflammations continue to be poorly understood, and an adequate classification scheme does not exist. Corticosteroids continue to be the preferred initial treatment, with the roles of radiation and nonsteroidal antiinflammatory medications to be determined. Specific causes of orbital inflammation such as Wegener granulomatosis must be considered to prevent potentially life-threatening complications. PMID- 10387482 TI - Eyelid cancers. AB - Eyelid cancers, like most malignancies, are on the rise, creating an ever enlarging population of patients with these diseases. Trends in eyelid cancer diagnosis, prognostic evaluation, prevention, and management are reviewed. Special emphasis is placed upon understanding perineural invasion by squamous cell carcinoma, the role of genetic mutations in eyelid cancer development and prognosis, and new techniques for total upper eyelid reconstruction. PMID- 10387483 TI - Aesthetic oculoplastic surgery. AB - Important concepts and advances continue to evolve in the field of aesthetic oculoplastic surgery. A keen interest in aesthetics exists, both on the part of the youth-oriented, aging baby-boomer population and on the part of many physicians and surgeons in a variety of different specialities. This review article highlights recent, important developments regarding cosmetic Botox injections for lines of facial expression, laser facial-resurfacing techniques, endoscopic and updated browlift techniques, lower-lid blepharoplasty with suborbicularis oculi fat lift, and subcutaneous Gore-Tex implants for treatment of deep facial rhytids. Because of the space limitations of this review, it is not possible to include a thorough step-by-step description of all surgical techniques. Rather, important concepts, references, and updates are emphasized. The information presented should be of interest to readers of varying levels of expertise. PMID- 10387484 TI - Orbital trauma. AB - In this review, a number of current issues on the management of orbital trauma are discussed including the indications for orbital exploration in zygomatic complex fractures, the utility of transantral endoscopy in orbital trauma surgery, and preferred implant materials in orbital reconstruction. We emphasize the need for a classification system to define some of the clinical terms used in describing orbital trauma, to improve communication between clinical services, and to standardize the clinical literature on orbital trauma. PMID- 10387485 TI - Biomaterials in ophthalmic plastic and reconstructive surgery. AB - Over the past several years, new biomaterials have been developed or modified for use in ophthalmic plastic and reconstructive surgery. Hydroxyapatite and high density porous polyethylene are extensively utilized in enucleation and evisceration surgery as well as in orbit reconstruction. Absorbable plates and screws as well as cyanoacrylate glue are at the forefront of fracture repair. Silicone and polytetrafluoroethylene eyelid implants are important adjuncts for adult and congenital ptosis repair. As technology advances, new materials will be developed and new applications identified. An understanding of biomaterials and their use is essential for every oculoplastic surgeon. PMID- 10387486 TI - Complications of postenucleation/evisceration implants. AB - Surgical correction of the anophthalmic socket is intended to alleviate a loss and fill a void both real and perceived. Despite advances in surgical technique and material, postoperative complications continue to occur at an unacceptable rate, from 10% to 28% depending on the series. This review details the complications of the various implants and the efficacy of the proposed management of said complications. The discussion is limited to a review of literature in the past year. PMID- 10387487 TI - Serum lipid concentrations correlate with the progression of chronic renal failure. AB - OBJECTIVE: To explore the distribution pattern for serum lipid concentrations among patients with different degrees of chronic renal failure; to study the characteristics of abnormal lipid metabolism for chronic renal failure patients when the disease progress further. SETTING: No. 255 Hospital of PLA, Tangshan, Hebei, China; No. 281 Hospital of PLA, Beidanhe, Hebei, China; and the General Hospital of Beijing Military Region, Beijing, China. PRACTICE DESCRIPTION: A total of 240 serum/urine samples from 50 healthy volunteers and from 190 patients with different degrees of chronic renal failure, which fall into four groups according to their glomerular filtration rates, were measured for serum levels of triglyceride, lipoprotein(a), lipoprotein(a) cholesterol, total cholesterol, apolipoprotein A1, apolipoprotein B100, low density lipoprotein cholesterol, high density lipoprotein cholesterol, and for urine albumin concentrations; the levels of these criteria were compared between the control group and diseased groups; the mean concentrations of different lipid variables were paired and subjected to linear regression analysis. MAIN OUTCOME MEASUREMENTS: Glomerular filtration rates were estimated by the iohexol clearance method, in which plasma content of iohexol was measured with high performance liquid chromatography; concentrations of triglyceride, lipoprotein(a), lipoprotein(a) cholesterol, total cholesterol, apolipoprotein A1, apolipoprotein B100, low density lipoprotein cholesterol, high density lipoprotein cholesterol, and albumin were assayed according to standard protocols. RESULTS: Serum levels of triglyceride, lipoprotein(a), lipoprotein(a) cholesterol, total cholesterol, apolipoprotein A1, apolipoprotein B100, low density lipoprotein cholesterol, and urine albumin contents were significantly higher, whereas those of high density lipoprotein cholesterol were lower, in diseased groups than that of the control (p < 0.05, p < 0.01). When the disease progressed, concentrations of these criteria increased or decreased further (p < 0.01, p < 0.05). Significant correlations were found between a few lipid criteria for their mean concentrations in diseased groups. CONCLUSION: The study demonstrates a correlation between abnormalities of lipid metabolism and the degrees of kidney insufficiency, and a correlation within certain kinds of lipid criteria in patients with different degrees of renal damage. The results suggest the existence of multi-correlations in vivo in catabolism and metabolism of lipid, lipoprotein, apolipoprotein, and protein in the patients. The exact mechanism responsible for the association and correlation remains to be clarified. PMID- 10387488 TI - Autoimmune hemolytic anemias: characteristics and classification. AB - AIHAs may be caused by either IgG or IgM class antibodies which affects the presentation and severity of the hemolytic process. In addition, the IHA may be idiopathic or secondary in etiology. Resolution of underlying diseases in the secondary type will often resolve the hemolysis; however, treatment may require more than one course of action, especially in the WAIHAs. PMID- 10387489 TI - Drug-induced hemolytic anemias: increasing complications to therapeutic interventions. AB - There are recent reports of severe drug-induced immune hemolysis caused by several different classes of drugs. Second and third generation cephalosporins, diclofenac, fludarabine, carboplatin, and beta-lactamase inhibitors are among the drugs associated with severe or fatal hemolysis. Studies on patients who exhibit hemolysis after ingesting these drugs indicate that the four classical mechanisms of drug-induced hemolytic anemia may overlap. These studies appear to support the unified theory for induction of drug-induced immune hemolytic anemia. PMID- 10387490 TI - Laboratory investigation of autoimmune hemolytic anemias. AB - The investigation of suspected autoimmune hemolytic anemias includes not only the laboratory investigation but also the patient's presenting symptoms and a complete medical history. A drug history is especially important if drug-induced hemolytic anemia is suspected. The patient's direct antiglobulin test results, both polyspecific and monospecific, guide the remainder of the workup. Serum and eluate testing may include untreated reagent red cells with and without the presence of unbound drug(s) and drug-treated reagent red cells. If autoantibodies are demonstrable in the serum, removal of those autoantibodies and subsequent testing for underlying alloantibodies is essential in the provision of safe blood for transfusion. PMID- 10387491 TI - Human influenza: viral mutations and altered tropisms. AB - Influenza is a virus that is capable of causing a pandemic of the human race. Influenza has the ability to infect humans by mutating and altering its pathogenic characteristics. Efforts must be made worldwide to educate people about the possibilities of a potential outbreak. Awareness of optimal conditions which could lead to viral mutation and human to human transmission of a neogenetic strain of influenza appears to be a key deterrent against future cases. PMID- 10387492 TI - Ethical responsibility. American Society for Clinical Laboratory Science. AB - In order to acknowledge the rights of the patient and the needs of the laboratory within a facility, ASCLS believes that telling the truth and respect for individuals be guiding principles. We acknowledge that this will not be easy; old protective habits of institutions and individuals die hard. Some may be unwilling to pay the price for abiding by these principles and will take another view. And this is their choice. The choice of ethical systems is, after all, the right of the individual. PMID- 10387494 TI - Clinical laboratory scientists' view of the competencies needed for current practice. AB - OBJECTIVE: To provide a current description of competent clinical laboratory scientists (CLSs) that can be used as a guide for educators, practitioners, and students. DESIGN: A survey of clinical laboratory science (CLS) practitioners was developed to assess current work settings and important competencies in those settings. The survey also addressed graduate school enrollment and the impact of multi-skilling on current practice. SETTINGS AND PARTICIPANTS: 135 graduates of the Division of Clinical Laboratory Science at the University of North Carolina at Chapel Hill from 1987 to 1996. MAIN OUTCOME MEASURES: Participants' responses to questions about their current job titles, clinical specialty, type of institution, multi-skilling, graduate school enrollment, and the competencies they considered important for their jobs were analyzed to provide a description of current practice. RESULTS: The response rate for the survey was 73%. The majority of the respondents were employed as staff CLSs in medium to large hospitals. Thirteen percent of the respondents indicated that they had graduate degrees and an additional 13% were currently enrolled in a graduate or professional program. Fourteen percent of the respondents reported that they were working in the laboratory profession and were performing some health care skills not included in the CLS program. The graduates described 15 major areas of competence important for current clinical practice. The skill or competency mentioned most frequently by the respondents was interpersonal skills followed by flexibility. Competence in technical and scientific skills was ranked third and problem solving abilities was ranked fourth. CONCLUSION: The results of this study describe a CLS practitioner who is able to communicate well with others as a team member; flexible and open to change in the work environment; technically component; able to solve problems and correlate clinical information; organized; and involved in the management and leadership of the clinical laboratory. This description can help educators design curricula, guide practitioners' self assessment, and inform students who are considering a career in CLS. PMID- 10387493 TI - Turnaround times in the laboratory: a review of the literature. AB - OBJECTIVE: To conduct a review of the literature for current information pertaining to turnaround times in the clinical laboratory. To evaluate the methods previously used for improving turnaround times and provide a reference for laboratories. DESIGN: The literature was reviewed for information related to turnaround times in the laboratory. Information was limited to literature published from 1989 to present in order to cover the more technological innovations. RESULTS: Several methods have been used in recent years for the improvement of turnaround times in the clinical laboratory. Among these are pneumatic tubing systems, satellite laboratories, point-of-care testing, and computer technology. CONCLUSIONS: There is still the need for faster turnaround times. Technological advances are enabling laboratories to better meet these needs leading to improved user satisfaction. PMID- 10387495 TI - Health and the Celtic tiger: progress of health promotion in modern Ireland. PMID- 10387496 TI - Formative research for developing targeted skin cancer prevention programs for children in multiethnic Hawaii. AB - Skin cancer is a significant and increasing public health problem. Improvement in sun protection practices among children holds great promise for prevention, and parents and caregivers play important roles. Health promotion programs are most likely to succeed when based on a systematic planning process including an understanding of current practices, beliefs, social norms and environments. This article describes formative research used to help develop the SunSmart skin cancer prevention program in Hawaii. Group discussions and interviews were conducted with 216 children in grades 1, 2 and 3, 15 parents, and 27 recreation staff. Children's discussion groups took place in intact classrooms. A combination of quantitative and qualitative methods was used. Multiple raters and an iterative process were used to analyze data from survey forms, observer impressions and audio tapes, and to draw the main conclusions. Sun protection practices in all groups were inconsistent, though general awareness about prevention was widespread. Children reported a reluctance to cover up with long pants and sleeves, and wide-brim hats, and did not understand what skin cancer was. Parents and recreation staff were supportive of education and policy supports, to improve both their own and the children's prevention habits. They were enthusiastic about interactive and creative activities. We conclude that targeted skin cancer prevention messages and strategies for Hawaii's children should promote gradual changes, provide environmental supports, and involve parents and recreation staff. Both the findings and procedures have implications for prevention elsewhere. PMID- 10387497 TI - The use of observational methods for monitoring sun-protection activities in schools. AB - Evaluation of health promotion interventions aimed at behavioural or environmental change involves assessing change that occurs as a result of the program. Direct observational methods can be used for this purpose and this paper describes three such methods that we pilot tested for use in a 5-year intervention study aimed at reducing sun exposure in primary school children. (1) Monitoring 'No hat, no play' policies. This method involved video taping children in selected school play areas during lunch time and analysing the content of the videos to assess the proportion of children wearing various types of hats. (2) Assessing shade provision in the playground. This method involved taking aerial photographs of each school and using them to estimate the proportion of shade in play areas available to children at lunchtime. (3) Shade use. This involved children wearing polysulphone film badges to measure the amount of UV-B exposure they received during one lunch period, relative to the total possible dose registered on index badges. Each method was implemented successfully, and we demonstrated that the video and aerial photography methods produced highly reproducible results and that all three methods were feasible. These three methods will be used in our intervention study to assess longitudinal change in schools' sun-protection policy and practice. PMID- 10387498 TI - Transgender HIV prevention: implementation and evaluation of a workshop. AB - Virtually no HIV prevention education has specifically targeted the transgender community. To fill this void, a transgender HIV prevention workshop was developed, implemented and evaluated. A 4 h workshop, grounded in the Health Belief Model and the Eroticizing Safer Sex approach, combined lectures, videos, a panel, discussion, roleplay and exercises. Evaluation using a pre-, post- and follow-up test design showed an increase in knowledge and an initial increase in positive attitudes that diminished over time. Due to the small sample size (N = 59) and limited frequency of risk behavior, a significant decrease in unsafe sexual or needle practices could not be demonstrated. However, findings suggested an increase in safer sexual behaviors such as (mutual) masturbation. Peer support improved significantly. Future prevention education should make special efforts to target the more difficult-to-reach, high-risk subgroups of the transgender population. PMID- 10387499 TI - Knowledge, risk perception of AIDS and reported sexual behaviour among students in secondary schools and colleges in Tanzania. AB - A questionnaire survey was carried out among 1041 students in secondary schools and colleges in Dar-es-Salaam, Tanzania to evaluate the relationship between HIV risky sexual behaviour and anti-condom bias, as well as with AIDS-related information, knowledge, perceptions and attitudes. Self-reportedly, 54% of students (75% of the boys and 40% of the girls) were sexually active, 39% had a regular sexual partner and 13% had multiple partners in the previous year. The condom use rate was higher than previous reports. However, 30% of sexually active respondents did not always use condoms (Risk-1 behaviour) and 35% of those with multiple partners in the previous year did not always use condoms (Risk-2 behaviour). Multiple logistic regression analyses indicated that 'sex partner hates condom' had association with both Risk-1 behaviour (OR 2.47; 95% CI 1.58 3.85) and Risk-2 behaviour (OR 2.47; 95% CI 1.10-5.48). 'Use of condom prevents HIV infection' also had association with both Risk-1 behaviour (OR 2.09; 95% CI 1.19-3.67) and Risk-2 behaviour (OR 3.73; 95% CI 1.28-11.03). Students engaging in risky behaviour were aware of the risk, even though they failed to change their behaviour. Reasons for the AIDS epidemic among Tanzanian students and the importance of more effective AIDS education are also discussed. PMID- 10387500 TI - 'Fatalism', accident causation and prevention: issues for health promotion from an exploratory study in a Yoruba town, Nigeria. AB - As countries experience the 'epidemiological transition' with a relative decline in infectious diseases, accident rates tend to increase, particularly road traffic accidents. The health promotion interventions intended to prevent or minimize the consequences of accidents have been developed in predominantly Western, industrialized countries. Although some of these solutions have been applied with success to less developed countries, there are also good reasons why such solutions are ineffective when tried in a different context. Health promotion as developed in the West has a particular ideological bias, being framed within a secular, individualist and rationalist culture. Different cosmologies exist outside this culture, often described as 'fatalist' by Western commentators and as obstructing change. Changing these cosmologies or worldviews may not fit with the ethic of paying due respect to the cultural traditions of the 'target group'. Health promotion is therefore faced with a dilemma. In addition to different worldviews, the different levels of development also mean that solutions formulated in richer countries do not suit poorer countries. This paper uses a small exploratory study in a Yoruba town in Nigeria to examine these points. Interviews with key informants were held in March 1994 in Igbo-Ora and data were extracted from hospital records. Levels of accidents from available records are noted and people's ideas about accident prevention are discussed. Recommendations as to the way forward are then proposed. PMID- 10387501 TI - Predictors of the prevalence of tobacco use among Francophones and Anglophones in the province of Ontario. AB - This study examines Francophone/Anglophone differences in levels and patterns of tobacco use and associated risk factors in the province of Ontario, Canada. Estimates are derived from the self-administered portion of the 1990 Ontario Health Survey, a random probability survey of Ontario residents. The sample consists of 1127 Francophones and a random subset of 4023 Anglophones. Evidence, unique to Francophones, indicates a steady age-related decline in the median age of onset of daily cigarette consumption. Unlike Anglophones, multivariate results reveal that Francophones age 35-44 are significantly more likely than all other age groups to smoke cigarettes daily and to smoke a pack or more daily. Sub groups within Ontario's Francophone community may be experiencing health-related risks associated with frequent and heavy consumption of cigarettes, and thus may be in need of addiction-related services. PMID- 10387502 TI - Sport activity in adolescence: associations with health perceptions and experimental behaviours. AB - Despite the relevance of this research topic from a public health perspective, there is currently a lack of objective data on European adolescents' sport activity, notably the associations between their sport habits and their health attitudes and behaviours, which may have important consequences both in terms of somatic (cardiovascular) health and mental health. The objective of the present study was to determine the direction and strength of the associations between the frequency of sport and health variables; in particular, perceptions of health, self image, substance use and experimental behaviours. Data were collected as part of the 1993 Swiss Multicentric Adolescent Survey on Health. In this survey, anonymous self-administered questionnaires were distributed to a national representative sample of 10,000 in-school adolescents (15-20 years of age). Univariate analyses explored the relationships between the level of sport activity and health variables; then logistic regression analyses examined the strength of these relationships. According to the results, half of the sample do sports more than twice a week, boys more often as part of a sports club. Differences between non-athletic and athletic adolescents describe the latter as having less somatic complaints, more confidence in their future health, a better body image, a lesser tendency to attempt suicide, a higher frequency of use of the car seat belt, and a lower use of tobacco, wine and marijuana. Links between the frequency of sport activity and the locus of control related to health, general satisfaction with life or sexual behaviours are less strong. It must be noticed that the cross-sectional data collection precludes the establishment of a causal relationship between exercise and health behaviours. However, the existing links underline the coexistence of positive health characteristics and sport activity, suggesting that an incitement to get involved in physical activity may be a necessary component of a comprehensive prevention approach among adolescents. PMID- 10387503 TI - A method in search of a theory: peer education and health promotion. AB - Peer education has grown in popularity and practice in recent years in the field of health promotion. However, advocates of peer education rarely make reference to theories in their rationale for particular projects. In this paper the authors review a selection of commonly cited theories, and examine to what extent they have value and relevance to peer education in health promotion. Beginning from an identification of 10 claims made for peer education, each theory is examined in terms of the scope of the theory and evidence to support it in practice. The authors conclude that, whilst most theories have something to offer towards an explanation of why peer education might be effective, most theories are limited in scope and there is little empirical evidence in health promotion practice to support them. Peer education would seem to be a method in search of a theory rather than the application of theory to practice. PMID- 10387504 TI - Computer-tailored nutrition education: differences between two interventions. AB - The impact of two computer-tailored nutrition education interventions was assessed and compared in a randomized trial among 315 subjects with a pre-test post-test comparison group design. Respondents in both the experimental and the comparison group received feedback tailored to their consumption of fat, fruit and vegetables. Respondents in the experimental group received additional psychosocial feedback tailored to their attitudes, perceived social support and self-efficacy expectations towards reducing their fat consumption and increasing their consumption of fruit and vegetables. A significant reduction in fat consumption and increase in the consumption of fruit and vegetables were found in both the experimental and the comparison group between pre-test and post-test. Respondents in the experimental group more often indicated that the feedback they received was interesting and easy to understand. Respondents in the comparison group more often reported having reduced their fat consumption because of the feedback they received. No significant differences in consumption of fat, fruit and vegetables were found at post-test between the experimental group and the comparison group. These results do not support the hypothesis that additional psychosocial information is an essential component of effective tailored feedback. The results indicate that tailored feedback might be effective in inducing dietary changes. PMID- 10387505 TI - A tailored multimedia nutrition education pilot program for low-income women receiving food assistance. AB - This article describes the development and pilot evaluation of a tailored multimedia program to improve dietary behavior among 378 low-income women enrolled in the Food Stamp program in Durham, North Carolina. After randomization to intervention or control groups, participants completed a baseline survey and were resurveyed 1-3 months post-intervention. Measures included dietary fat intake assessed using a brief food-frequency questionnaire, stage of change, knowledge of low-fat foods, self-efficacy and eating behavior questions. The computer-based intervention consisted of a tailored soap opera and interactive 'info-mercials' that provided individualized feedback about dietary fat intake, knowledge and strategies for lowering fat based on stage of change. At follow-up, intervention group participants had improved significantly in knowledge (P < 0.001), stage of change (P < 0.05) and certain eating behaviors (P < 0.05) compared to the control group. Both study groups had lowered their reported fat intake markedly at follow-up (P < 0.001), but did not differ significantly from each other. A majority of participants rated the program as very helpful and were interested in using a similar program in the future. The findings of this pilot study suggest that computerized tailored self-help health promotion programs may be effective educational interventions for lower income and minority populations. PMID- 10387506 TI - Health promotion in couples adapting to a shared lifestyle. AB - In a pilot health promotion program for couples, we aimed to build on re evaluation of attitudes to health occurring early in marriage, and social support provided by partners, to address the weight gain and physical inactivity which may follow marriage. A randomized controlled trial lasting 16 weeks used six modules focusing on nutrition and physical activity but including information about alcohol and smoking. Thirty-four of 39 couples enrolled completed the study. Self-efficacy for diet and physical activity increased significantly in the program group while ranking of barriers to healthy behaviours decreased and ranking of beliefs about the benefits of health behaviours increased relative to controls. Intake of fat, take-away foods and alcohol decreased, and consumption of fruit, vegetables and reduced-fat foods increased significantly in the program group. Physical activity in the program group increased by the equivalent of 50 min of brisk walking weekly but did not differ significantly from controls. Cholesterol fell significantly by 6% more in the program group than controls. In focus groups, participants unanimously found the program valuable. Health promotion programs designed for couples can achieve short-term changes in behaviour and risk factors. Larger trials with longer-term monitoring, incorporating feedback from focus groups and cost-benefit analysis, are in progress. PMID- 10387507 TI - Key informant surveys as a tool to implement and evaluate physical activity interventions in the community. AB - Key informant surveys are important tools for planning and evaluating community health programs. A survey was conducted to gather views on policies toward physical activity from four sets of key informants: physicians, church leaders, business leaders and civic leaders. Surveys were mailed to 797 key informants who were selected from 12 southeastern Missouri counties. For comparison, data from a telephone survey of 2106 persons in the general population were also analyzed. The majority (> 85%) in all four key informant groups were very supportive of required physical education in schools, but less supportive (< 69%) of government funding for places where community members can exercise. Physicians perceived community members as having somewhat greater access to places to exercise relative to the other key informant groups. Comparisons of the key informant surveys to the population survey indicated similar levels of support for physical activity policy. The information from this survey has been useful in identifying support for physical activity policy and gaining access to potential influences for community change. Since key informant research in the area of physical activity policy and cardiovascular disease prevention is sparse, there is a need for future studies. PMID- 10387508 TI - Talking trash. Health care's got a radical new attitude about waste disposal. PMID- 10387509 TI - Wired e-commerce is booming. Now it's about to burst onto the health care scene- or so we've been hearing. PMID- 10387510 TI - Round 'em up. PMID- 10387511 TI - You can get there from here. PMID- 10387512 TI - Befriend a doc, gain an ally. PMID- 10387513 TI - Stats. The managed care challenge. Don't let reduced revenue put you in the red. PMID- 10387514 TI - Problems for everyone in IT staffing needs. PMID- 10387515 TI - Zero tolerance for billing errors. PMID- 10387516 TI - Rethinking the CPR: is perfect the enemy of the good? PMID- 10387517 TI - The key to IS technology: re-engineering versus replacement. PMID- 10387518 TI - Workflow tools cut costs for high quality care. PMID- 10387519 TI - Physician data entry is the solution. PMID- 10387520 TI - What works. Wireless networks allow unprecedented response time. PMID- 10387521 TI - What works. ATM network increases performance: services for medical staff and patients are available without delay. PMID- 10387523 TI - HotList: CPR/EMR. PMID- 10387522 TI - What works. Data warehouse: decision support solution reduces patient admissions, saves payer millions. PMID- 10387524 TI - Focusing on the patient: keeping peripheral vision on organization. PMID- 10387525 TI - GenXers: will the workplace ever be the same? PMID- 10387526 TI - Erase the battle lines: how to cut out conflicts with MCO case managers. AB - With managed care penetration increasing, it's more important than ever for hospital case managers to find ways to resolve the inevitable conflicts that arise with their managed care-based counterparts. Typical conflicts include struggles over authorization, vendor selection, lack of contact, and access to the patient. Some conflicts can be resolved simply by increasing the level of communication--usually by having managed care case managers stationed in the hospital itself. But even when contact is only by telephone, there are steps you can take to ease the tension. One way is simply to keep managed care case managers informed regarding such things as return admissions by problem patients. Effective discharge planning practices also can strengthen bonds, especially when it comes to patients with complex care needs. PMID- 10387527 TI - CMs fight for acceptance of premature infant path. AB - Responding to benchmarking data indicating that their length of stay for premature infants was above the national average, case managers at University Hospital-University of Colorado Health Sciences Center in Denver formed a multidisciplinary team to develop a pathway designed to address the problem. But they encountered stiff initial resistance from both staff nurses and physicians, many of whom regarded the pathway as "cookbook medicine." The main appeal to nurses was that the pathway's preprinted orders would help standardize care--a key selling point at this teaching hospital, where attending physicians and interns rotate in and out on a monthly basis. Case managers also changed the pathway's documentation requirements at the suggestion of the nurses, adopting a charting-by-exception approach. Physicians have been slower to come around, but the case managers are optimistic that outcomes data on the revised pathway, due within six months, will demonstrate the pathway's benefits. PMID- 10387528 TI - Continuum improves CHF patient education. PMID- 10387529 TI - Hospital brings social work into case management fold. AB - To prepare for a wave of managed care penetration and to reduce duplication of work, 240-bed Jefferson Memorial Hospital in Crystal City, MO, integrated its social work and utilization review departments. The resulting case management services department features seven case coordinators and three social workers who often work out individual responsibilities for a given case among themselves. The newly hired case coordinators received extensive training in utilization review and discharge planning. The training curriculum involved trips to home health, hospice, a rehabilitation hospital, and a larger case management department. PMID- 10387530 TI - JCAHO's proposed rules on clinical practice guidelines. PMID- 10387531 TI - Congress approves FY 2000 budget, sets aside money for Medicare reform. PMID- 10387532 TI - Coalition seeks additional exclusions from SNF financial responsibility. PMID- 10387533 TI - Medicare claims now required to be Y2K compliant. PMID- 10387534 TI - Subacute SNFs struggle to meet demands of Medicare PPS uncertainty of ground rules, payment cuts pose biggest challenges. Part II: Managing costs, MDS scheduling, PPS billing, and the future. PMID- 10387535 TI - Sun Healthcare incurs $754 million loss for 1998. PMID- 10387536 TI - The media, the public and the inside story. PMID- 10387537 TI - The future of ethical thinking in healthcare management. PMID- 10387538 TI - Stereotactic breast biopsy units in the United States. PMID- 10387539 TI - Year around management magic. PMID- 10387540 TI - From prophets to profits. PMID- 10387541 TI - Disease prevention among postmenopausal women in the managed care setting. Based on a presentation by Neal M. Friedman, MD. PMID- 10387542 TI - The role of the ECHO model in outcomes research and clinical practice improvement. AB - In the articles that follow in this Special Report supplement to The American Journal of Managed Care, the reader will note that various types of outcomes- humanistic and economic, as well as clinical--are measured to achieve a balanced picture of the comprehensive impact of the healthcare interventions implemented. Clinical research has only recently evolved to a state in which a balanced systems approach to outcomes measurement is moving to the forefront. It is therefore important to understand the historical context of the outcomes movement within which the multidimensional approach developed. A description of this historical context is provided below, followed by an overview of the ECHO (Economic, Clinical, and Humanistic Outcomes) Model and case examples of the balance-of-outcomes model as implemented in a large integrated delivery system. PMID- 10387543 TI - Economic outcomes of a targeted intervention program: the costs of treating allergic rhinitis patients. AB - OBJECTIVES: To determine the annual costs of treating allergic rhinitis patients in a managed care environment and to assess the effect of a treatment intervention program on direct and indirect costs. DESIGN: Two arms of an economics study were designed to calculate annual costs of treating allergic rhinitis in Lovelace Health Systems. Direct and indirect costs were also reviewed for patients participating in an intervention program designed to improve patient outcomes during the 1996 fall allergy season. PATIENTS AND METHODS: Annual medical costs of treating allergic rhinitis within the Lovelace system were reviewed using a patient database. A total of 7936 patients with allergic rhinitis symptoms were identified in the database using a case-finding algorithm. An equal number of patients without allergy conditions were selected for the comparison group. In addition to calculating annual costs of treating allergic rhinitis, direct and indirect costs were reviewed for patients participating in a rhinitis intervention program to determine differences in cost between the treatment and control groups. An intervention group of 247 patients was selected to receive care at clinics randomized to use practice guidelines to improve treatment, while 255 patients were treated in the control group clinics, which did not alter treatment practices. RESULTS: Annual expenditures were nearly $2 million more for the allergic rhinitis group than for the control group. In the intervention study, treatment and control groups expended the same in direct costs, but the intervention group showed a trend toward decreased indirect costs. CONCLUSION: Costs of allergic rhinitis are not trivial to a managed care organization; a specifically designed intervention program shows potential for minimizing the costs associated with the ailment. PMID- 10387544 TI - Diagnosis and treatment of allergic rhinitis: primary care in an integrated health system setting. AB - The first point of contact for many patients presenting with allergy symptoms is the primary care physician. And in the managed care system, this initial primary care visit is essential. Guidelines for the primary care physician in diagnosing and treatment rhinitis as well as referring patients to allergy specialists are described. In addition, optimal medication usage for treating mild, moderate, and severe rhinitis is detailed. PMID- 10387545 TI - Putting spotlight on HR saves system $400,000 a year. PMID- 10387546 TI - Bold new purchasing program to save NJ system $25 million. AB - Bold new purchasing program will be big cost saver for health system. Saint Barnabas Health Care System in West Orange, NJ, is withdrawing from its GPOs and taking advantage of its economies of scale by purchasing pharmaceutical and medical/surgical supplies on its own. Here's how Saint Barnabas created its own GPO. PMID- 10387547 TI - Top-performing physician groups focus on innovative cost control. AB - DATA BENCHMARKS: Top-performing physician group practices reveal why they are head and shoulders above their peers. A new study released by the Medical Group Management Association offers benchmark data that shows how leading group practices in various specialties are faring compared with their colleagues nationwide. PMID- 10387548 TI - Adding pediatric hospitalists speeds discharge process, improves continuity of care. AB - Pediatric hospitalists are an essential ingredient in some revamped pediatric units. Lehigh Valley Hospital in Allentown, PA, attributes reduced length of stay in its pediatrics unit in part to its use of a group of pediatric hospitalists. Find out the benefits of such a program and how it works. PMID- 10387549 TI - Use of digital technology improves documentation and decreases costs in radiation oncology. PMID- 10387550 TI - Advances in patient positioning. AB - New technologies and processes have allowed for advances in patient positioning. Indexed immobilization allows clinicians to enhance treatment effectiveness and maximize clinical efficiency. When the same patient positioning system and couch are used in simulation and in the treatment room it is feasible to ensure identical patient setups between simulation and treatment. The emergence of 3-D conformal therapy and IMRT have lead the radiotherapy community to place increased importance and need for accurate patient positioning. As a result, radiotherapy vendors, as well as many radiation oncology departments, have developed their own immobilization devices to reduce patient setup time and ensure exact repeat positioning. Indexed immobilization is intended to facilitate precise repeatable patient positioning both in moving from simulation to treatment and in standardizing day-to-day set-ups. In addition to precise patient positioning, indexed immobilization can significantly reduce the length of treatment sessions by reducing patient movement and by standardizing the placement and set-up of immobilization devices. Coupled with the use of 3D treatment planning capability, precise positioning can facilitate treatments for flat or wedged conformal therapy and Intensity Modulated Radiation Therapy (IMRT). This article summarizes the features and operational benefits of indexed immobilization and then outlines the implications for clinical and departmental processes. The most significant benefits come when the user's processes are modified to take advantage of the enabling technology. PMID- 10387552 TI - Management zones--are they safe? PMID- 10387551 TI - Palliative radiotherapy for skeletal metastases: cost-substitution analyses and economic impact. PMID- 10387553 TI - Medicare solvent until 2015, trustees say: key senators set to introduce reform legislation. PMID- 10387554 TI - Feds kick fraud and abuse program into high gear: holder defends False Claims Act guidelines. PMID- 10387555 TI - Bipartisanship is key to health care reform. PMID- 10387556 TI - The broken rhetoric of the Medicare Trust Fund debate. PMID- 10387557 TI - Tobacco settlements: an update. PMID- 10387558 TI - Privacy legislation hits Capitol Hill radar screen. PMID- 10387559 TI - Career advancement for rising hospital executives. PMID- 10387560 TI - A look to the future. Home health PPS. AB - HCFA continues to move toward its goal of implementing a per-episode prospective payment system for home health care. What is in store for agencies, and how can they be prepared for all of the changes that will come when the shift from the current interim payment system to the new model is complete? If they are to thrive, agencies must understand the background behind IPS and PPS and what the future holds. PMID- 10387561 TI - The legal considerations surrounding OASIS (Outcome Assessment and Information Set) reporting requirements. AB - Effective February 24, 1999, all home health agencies that participate in the Medicare program implemented comprehensive assessments that contain a core data set known as OASIS for all adult, nonmaternity patients. The completion of the comprehensive assessment, using OASIS, also requires agencies to report OASIS data to designated regulatory bodies at least every 30 days. Both requirements are new Conditions of Participation for the Medicare program. PMID- 10387562 TI - Home health medical review: responding to requests for information. PMID- 10387563 TI - Overpayment guidance for home health agencies. AB - IPS brought many changes for home health providers, including intensified claims review and audits, resulting in overpayments. Agencies have several options despite the limited protection they have under Medicare, including challenging determinations, rebutting the decision, requesting a waiver, and appealing decisions. PMID- 10387564 TI - OIG releases compliance guidelines for HME providers. PMID- 10387565 TI - Mergers, affiliations and networks: the biggest challenge. PMID- 10387566 TI - OASIS (Outcome and Assessment Information Set) requirements need refining. PMID- 10387568 TI - The role of the notified body in the CE marking of medical devices. PMID- 10387567 TI - Care and trembling. AB - Medicare's antifraud program and its stepped-up focus on home health care have created a climate of intimidation, fear, and guilt. Increasingly, paid caregivers feel that this climate contradicts their sense of common decency and morality, makes them feel like criminals for caring as they watch their clients get hurt by the system. What hurts them most is that they are forced to participate in this withdrawal of care. This discussion showcases the dangers of making caring work formal and public. PMID- 10387569 TI - Natural and mechanical smoke control systems. PMID- 10387570 TI - Year 2000 project map. PMID- 10387571 TI - Special twin socket for IT equipment. PMID- 10387572 TI - Electromagnetic interference problems in the healthcare environment. PMID- 10387573 TI - Firing up the front line. AB - For many organizations, achieving competitive advantage means eliciting superior performance from employees on the front line--the burger flippers, hotel room cleaners, and baggage handlers whose work has an enormous effect on customers. That's no easy task. Front line workers are paid low wages, have scant hope of advancement, and--not surprisingly--often care little about the company's performance. But then how do some companies succeed in engaging the emotional energy of rank-and-file workers? A team of researchers at McKinsey & Company and the Conference Board recently explored that question and discovered that one highly effective route is demonstrated by the U.S. Marine Corps. The Marines' approach to motivation follows the "mission, values, and pride" path, which researchers say is practical and relevant for the business world. More specifically, the authors say the Marines follow five practices: they over-invest in cultivating core value; prepare every person to lead, including front line supervisors; learn when to create teams and when to create single-leader work groups; attend to all employees, not just the top half; and encourage self discipline as a way of building pride. The authors admit there are critical differences between the Marines and most businesses. But using vivid examples from companies such as KFC and Marriott International, the authors illustrate how the Marines' approach can be translated for corporate use. Sometimes, the authors maintain, minor changes in a company's standard operating procedure can have a powerful effect on front line pride and can result in substantial payoffs in company performance. PMID- 10387574 TI - From spare change to real change. The social sector as beta site for business innovation. AB - Corporations are continually looking for new sources of innovation. Today several leading companies are beginning to find inspiration in an unexpected place: the social sector. That includes public schools, welfare-to-work programs, and the inner city. Indeed, a new paradigm for innovation is emerging: a partnership between private enterprise and public interest that produces profitable and sustainable change for both sides. In this article, the author shows how some companies are moving beyond corporate social responsibility to corporate social innovation. Traditionally, companies viewed the social sector as a dumping ground for their spare cash, obsolete equipment, and tired executives. But that mind-set hardly created lasting change. Now companies are viewing community needs as opportunities to develop ideas and demonstrate business technologies; find and serve new markets; and solve long-standing business problems. They focus on inventing sophisticated solutions through a hands-on approach. This is not charity; it is R & D, a strategic business investment. The author concedes that it isn't easy to make the new paradigm work. But she has found that successful private-public partnerships share six characteristics: a clear business agenda, strong partners committed to change, investment by both parties, rootedness in the user community, links to other organizations, and a commitment to sustain and replicate the results. Drawing on examples of successful companies such as IBM and Bell Atlantic, the author illustrates how this paradigm has produced innovations that have both business and community payoffs. PMID- 10387575 TI - The smart-talk trap. AB - In today's business world, there's no shortage of know-how. When companies get into trouble, their executives have vast resources at their disposal: their own experiences, colleagues' ideas, reams of computer-generated data, thousands of publications, and consultants armed with the latest managerial concepts and tools. But all too often, even with all that knowledge floating around, companies are plagued with an inertia that comes from knowing too much and doing too little -a phenomenon the authors call the knowing-doing gap. The gap often can be traced to a basic human propensity: the willingness to let talk substitute for action. When confronted with a problem, people act as though discussing it, formulating decisions, and hashing out plans for action are the same as actually fixing it. And after researching organizations of all shapes and sizes, the authors concluded that a particular kind of talk is an especially insidious inhibitor of action: "smart talk." People who can engage in such talk generally sound confident and articulate; they can spout facts and may even have interesting ideas. But such people often exhibit the less benign aspects of smart talk as well: They focus on the negative, and they favor unnecessarily complex or abstract language. The former lapses into criticism for criticism's sake; the latter confuses people. Both tendencies can stop action in its tracks. How can you shut the smart-talk trap and close the knowing-doing gap? The authors lay out five methods that successful companies employ in order to translate the right kind of talk into intelligent action. PMID- 10387576 TI - A road map for natural capitalism. AB - No one would run a business without accounting for its capital outlays. Yet most companies overlook one major capital component--the value of the earth's ecosystem services. It is a staggering omission; recent calculations place the value of the earth's total ecosystem services--water storage, atmosphere regulation, climate control, and so on--at $33 trillion a year. Not accounting for those costs has led to waste on a grand scale. But now a few farsighted companies are finding powerful business opportunities in conserving resources on a similarly grand scale. They are embarking on a journey toward "natural capitalism," a journey that comprises four major shifts in business practices. The first stage involves dramatically increasing the productivity of natural resources, stretching them as much as 100 times further than they do today. In the second stage, companies adopt closed-loop production systems that yield no waste or toxicity. The third stage requires a fundamental change of business model--from one of selling products to one of delivering services. For example, a manufacturer would sell lighting services rather than lightbulbs, thus benefitting the seller and customer for developing extremely efficient, durable lightbulbs. The last stage involves reinvesting in natural capital to restore, sustain, and expand the planet's ecosystem. Because natural capitalism is both necessary and profitable, it will sub-sume traditional industrialism, the authors argue, just as industrialism sub-sumed agrarianism. And the companies that are furthest down the road will have the competitive edge. PMID- 10387577 TI - Will this open space work? AB - Northern Oil is moving offices, and CEO Fritz Schumacher wants to make the most of the move in this fictional case study. He believes that adopting an open-plan work space will reinvent how the company works, not to mention cut costs. Facilities manager Sasha Pasternak also supports the open plan. Her job would be easier, and her budget would stretch further, if Northern had standardized workstations and used partitions, not walls. And she likes the way the new design flattens the organization: everyone has the same amount of space and the same ergonomically sound furniture. The new building would have more conference rooms and just-in-time work spaces for employees who worked mostly off-site. And although she knew that initial meetings between the architects and Northern employees hadn't yielded much support for open space--people were attached to their private offices--she expected that people would warm to the idea. But when the new design was unveiled, employees were less than enthusiastic. They hurled questions like, How will workers concentrate if they can't shut their office doors? How will people have confidential meetings with their boss? And why would people stay at Northern when the competition offers them private offices? There was even talk of circulating a petition refusing to move to the new space. A week later, the architect presented revised plans to the project group. The new options would add costs and reduce the amount of space savings, but offering a choice to employees might make them feel less threatened. What should the project team do? Five commentators offer advice. PMID- 10387578 TI - Turning negotiation into a corporate capability. AB - Every company today exists in a complex web of relationships formed, one at a time, through negotiation. Purchasing and outsourcing contracts are negotiated with vendors. Marketing arrangements are negotiated with distributors. Product development agreements are negotiated with joint-venture partners. Taken together, the thousands of negotiations a typical company engages in have an enormous effect on both its strategy and its bottom line. But few companies think systematically about their negotiating activities as a whole. Instead they take a situational view, perceiving each negotiation to be a separate event with its own goals, tactics, and measures of success. Coordinating them all seems an overwhelming and impracticable job. In reality, the author argues, it is neither. A number of companies are successfully building coordinated negotiation capabilities by applying four broad changes in practice and perspective. First, they've established a company-wide negotiation infrastructure to apply the knowledge gained from forging past agreements to improve future ones. Second, they've broadened the measures they use to evaluate negotiators' performance beyond matters of cost and price. Third, they draw a clear distinction between the elements of an individual deal and the nature of the ongoing relationship between the parties. Fourth, they make their negotiators feel comfortable walking away from a deal when it's not in the company's best interests. These changes aren't radical steps. But taken together, they will let companies establish closer, more creative relationships with suppliers, customers, and other partners. PMID- 10387580 TI - How risky is your company? AB - In boom times, it is easy for managers to forget about risk. And not just financial risk, but organizational and operational risk as well. Now there's the risk exposure calculator, a new tool that will help managers determine exactly where and how much internal risk is mounting in their companies. The risk calculator is divided into three parts: The first set of "keys" alerts managers to the pressures that come from growth. Now that the company has taken off, are employees feeling increased pressure to perform? Is the company's infrastructure becoming overloaded? And are more new employees coming on board as the company rushes to fill positions? If the answer is yes to any one of those questions, then risk may be rising to dangerous levels. The second set of keys on the calculator highlights pressures that arise from corporate culture. Are too many rewards being given for entrepreneurial risk taking? Are executives becoming so resistant to bad news that no one feels comfortable alerting them to problems? And is the company's level of internal competition so high that employees see promotion as a zero-sum game? The final set of pressures, the author says, revolves around information management. When calculating these pressures, managers should ask themselves, what was the company's complexity, volume, and velocity of information a year ago? Have they risen? By how much? How much of the time am I doing the work that a computer system should be doing? High pressure on many or all of these points should set off alarm bells for managers. To control risk, managers have four levers of control at their disposal that will show them where they need to make organizational adjustments. PMID- 10387579 TI - Patching. Restitching business portfolios in dynamic markets. AB - In turbulent markets, businesses and opportunities are constantly falling out of alignment. New technologies and emerging markets create fresh opportunities. Converging markets produce more. And of course, some markets fade. In this landscape of continuous flux, it's more important to build corporate-level strategic processes that enable dynamic repositioning than it is to build any particular defensible position. That's why smart corporate strategists use patching, a process of mapping and remapping business units to create a shifting mix of highly focused, tightly aligned businesses that can respond to changing market opportunities. Patching is not just another name for reorganizing; patchers have a distinctive mindset. Traditional managers see structure as stable; patching managers believe structure is inherently temporary. Traditional managers set corporate strategy first, but patching managers keep the organization focused on the right set of business opportunities and let strategy emerge from individual businesses. Although the focus of patching is flexibility, the process itself follows a pattern. Patching changes are usually small in scale and made frequently. Patching should be done quickly; the emphasis is on getting the patch about right and fixing problems later. Patches should have a test drive before they're formalized but then be tightly scripted after they've been announced. And patching won't work without the right infrastructure: modular business units, fine-grained and complete unit-level metrics, and companywide compensation parity. The authors illustrate how patching works and point out some common stumbling blocks. PMID- 10387581 TI - Changing leaders: the board's role in CEO succession ... roundtable discussion. AB - The selection of a CEO is one of the most important- and risky-events in the life of any company. Yet the way CEOs are chosen remains little discussed and little understood. The succession process has traditionally unfolded behind closed doors -some observers have even likened it to the election of a pope. To shed light on what works and what doesn't in CEO succession, the authors lead a roundtable discussion with five distinguished corporate directors: Philip Caldwell, George D. Kennedy, G. G. Michelson, Henry Wendt, and Alfred M. Zeien. Collectively, the five directors have participated in dozens of successions, either as board members or as CEOs. In a lively and frank exchange of views and experiences, the roundtable participants explore a broad range of questions: What can a company do to ensure a successful succession? How should management-development and succession processes be managed? How should the board work with the sitting chief executive during the process? What makes for a strong CEO candidate? When should outside candidates be considered? How much competition should be encouraged among potential CEO candidates? What role should executive search firms play? What role should former CEOs play after they are succeeded? Their conversation illuminates a corporate challenge that is as difficult as it is important. PMID- 10387582 TI - Concurrent improvements in ambulatory cardiac catheterization practices following inpatient interventions. AB - Questions have been increasingly raised about the value of performing right heart catheterization. A preliminary analysis done in 1992 revealed significant interhospital variation in the frequency of the procedure among Medicare Part A and Medicaid patients in New York State, and it also suggested that the procedure was being performed routinely in some hospitals. In 1993, IPRO initiated a cooperative health care quality improvement program involving the state's 53 catheterization laboratories. As a result of this educational intervention, the rate of bilateral catheterization among Medicare Part A patients fell from 89/100,000 beneficiaries in 1992 to 65/100,000 in 1996, and the overall percentage of catheterized Medicare patients undergoing bilateral catheterization fell from 30.5% in 1992 to 17.4%. A major question was whether a corresponding decrease had occurred among ambulatory patients (Medicare Part B). To determine the answer, the Medicare Part B database was analyzed for the identical period of time. It was found that the percentage of ambulatory Medicare patients who underwent bilateral catheterization at the 53 laboratories fell from 37.6% in 1992 to 17.0% in 1996, paralleling the decline observed among inpatients. The results of this quality improvement study show that an educational intervention directed at inpatient practice patterns can have a similar impact on outpatient patterns. PMID- 10387583 TI - Primary care physicians' participation in managed care networks. AB - The study described in this article explored the relationships between primary care physician characteristics and patterns of managed care affiliation in a single region. Secondary data sources were used to investigate the affiliations of all primary care physicians in King County, Washington (Seattle and environs) with 29 managed care products in 1996. Descriptive findings indicate that specialty, board certification, and experience all are associated with the managed care affiliations held by physicians. Differences between managed care product provider lists suggest that there are different strategies for the design and management of provider networks. PMID- 10387584 TI - The prospective risk adjustment system. AB - The Episode Classification System is intended to perform two tasks. First, it will be a prospective capitation risk adjuster and predict future health care costs. It will do this by assigning each individual a single capitation risk adjustment category based on an analysis of the medical history and of health care services rendered during a specific period of time. Second, the Episode Classification System will create retrospective severity adjusted Episodes of illness or Episodes of Care. These latter Episodes will provide a framework for relating patient characteristics to the amount, type, and duration of services provided during the treatment of a specific disease. These Episodes will give users the ability to understand past costs and the risk of mortality. As such they will form the basis for provider profiling by allowing users to analyze a complete clinical episode. PMID- 10387585 TI - A way of obtaining isoresource consumption care episodes. AB - The purpose of this study is to identify groups of care episodes that involve pneumologic and cardiologic problems and that exhibit homogeneous patterns of frequentation and diagnostic test consumption in an ambulatory consulting room. A 1-year prospective study of care content was done, and the episodes were retrospectively analyzed 2 years later. Data were collected in an ambulatory cardiologic and pneumologic consulting room in Spain. Nonlinear principal components analysis was applied before cluster analysis. Five typologies with a homogeneous pattern of resource consumption were obtained: three related to acute episodes of care and two to chronic ones. PMID- 10387586 TI - Forging the link between health and human rights: celebrating over 50 years of the Universal Declaration of Human Rights. PMID- 10387587 TI - Assessing patient satisfaction across the continuum of ambulatory care: a revalidation and validation of care-specific surveys. AB - The need to develop reliable and valid measures of patient satisfaction in ambulatory care settings is underscored by the rapid growth and changes in this health care arena as well as the requirement to monitor and gauge quality of care. The purpose of this article is to provide evidence for the reliability and validity of patient satisfaction questionnaires designed specifically for three points of care across the ambulatory care continuum. These points are outpatient testing and physical therapy services, outpatient surgery, and home health care. The present effort represents an evaluation of revised questionnaires for the first two points and an initial assessment of the psychometric properties of the questionnaire for the third. The present results support the internal reliability and construct validity of each questionnaire. In addition, differences were found on select facility characteristics based on each questionnaire. PMID- 10387588 TI - Momma, oh Momma, I can't remember. AB - Institutionalized persons with dementia present many challenges to pastoral caregivers. The loss of memory and cognitive functions in these persons requires that caregivers value the immediate moment of pastoral contact and ground their ministry in relevant theological foundations. Examples of these challenges are provided and theological concepts are discussed. PMID- 10387589 TI - In memory of loved ones who have enriched our lives: helping staff create and hold memorial services in nursing homes. AB - This article describes the introduction of periodic memorial services in a nursing home for residents who died. The planning process as well as objections and concerns are discussed. A sample service is described, including the letter of invitation to family members and friends of deceased residents. The article concludes with three sample "tributes" to those whose lives were celebrated in a memorial service. PMID- 10387590 TI - "To see things as God sees them": theological reflections on pastoral care to persons with dementia. AB - Although theology is often seen as impractical speculation on unimportant matters, it serves as a necessary foundation--and provides valuable guidance--for chaplains who must provide pastoral care to persons with dementia and their families. Theology can help us "to see things as God sees them." Among the theological doctrines found in the Hebrew-Christian scriptures and traditions that are particularly helpful are the following: human creation "in the image of God"; human nature as a psychophysical unity; the dependence of all persons upon God's mercy; the centrality of community; and God's judgment of personal worth by standards very different from those of "the world." A model for applying these concepts and some thoughts of the importance of chaplains are offered. PMID- 10387591 TI - Spirituality, religion, and Alzheimer's disease. AB - The chaplain's ministry to persons with dementia, often of the Alzheimer's type, is vitally relevant to their clinical well-being. No chaplain should even think that because someone is demented, they can no longer be reached spiritually. While few scientific studies exist, clinical experience and anecdotal accounts suggest that selected pastoral interventions can enhance the quality of life of the mildly, moderately, and even severely demented individual. PMID- 10387592 TI - Pastoral care of problematic Alzheimer's disease and dementia affected residents in a long-term care setting. AB - Pastoral caregivers face many challenges in providing ministry to institutional persons with dementia. This article describes the psychosocial perspective of Bowlby concerning the management of persons with dementia and a pastoral care ministry based on it. Specific pastoral programs and interventions are described. The article contains four case studies and concludes with reflections concerning the chaplain's ministry. PMID- 10387593 TI - Pastoral care for the person with dementia. AB - We discuss the various stages of Alzheimer's disease and related disorders (ADRD) and present a psychosocial model which spiritual caregivers can use in their ministry, the Progressively Lowered Stress Threshold (PLST) model. We argue that religious activities are very important to these patients and that spiritual caregivers can make an important contribution. PMID- 10387594 TI - Forget me not: the spiritual care of people with Alzheimer's Disease. AB - This article observes that many clergy do not seem to understand the importance of ministry to persons with dementia. New understandings about the relationship between dementia and spirituality are presented and theological foundations explored. The article ends with a discussion of pastoral strategies that are important in this ministry. PMID- 10387595 TI - Assuring professional pastoral care for every nursing home resident. AB - Ministry to persons in nursing homes is built on two mandates: "... He has sent me to bring good news to the oppressed, to bind up the brokenhearted, to proclaim liberty to the captives, and release to the prisoners; ... to comfort all who mourn ..." (Isaiah 61:1-3). The federal government provides the second: "Quality of Life. A facility must care for its residents in a manner and in an environment that promotes maintenance or enhancement of each resident's quality of life" (OBRA '87, Guidance to Surveyors in Long Term Care Facilities, Code of Federal Regulations, Health Care Financing Administration, 1995, section 483.15, F240). This article discusses both the religious and the U.S. political history of caring for the old and frail. It concludes by describing political efforts in one state to increase the quality of that care and pastoral efforts to support the nursing assistants in long-term care facilities. PMID- 10387596 TI - The unique benefits of religious support during cardiac bypass surgery. AB - Compares the self-reports of family members waiting during the cardiac artery grafting surgery of a loved one and explores whether they make distinctions between the contributions of nonreligious and religious support. Results from regression analyses suggests that the use of religious sources of support was associated with both more positive religious and nonreligious psychosocial adjustment scores after the influences of nonreligious support were statistically removed. Notes that among the 13 religious support activities identified, family members reported using prayer most frequently. Concludes that using religious support sources to cope with this surgically related stress is associated with distinct subjective benefits beyond those contributed by nonreligious sources. PMID- 10387597 TI - Pastoral counseling and the changing times. AB - Recalls that although long-term psychotherapy was the underpinning of pastoral counseling in its early days of development and remains so for many practitioners today, recent outcome studies demonstrate that brief counseling is what in fact counselees prefer and is as effective as long-term therapy. It is what congregational pastors traditionally practiced for generations. Suggests that pastoral care-givers practice short-term counseling, arguing that the dissonance that occurs when one believes in the superiority of long-term counseling but nevertheless engages in primarily short-term counseling disrupts therapeutic ends. Proposes that brief pastoral counseling needs to be the model of choice for contemporary pastoral counselors, not only for practical reasons but for moral reasons as well. PMID- 10387598 TI - Pastoral care and the market economy: time-limited psychotherapy, managed care, and the pastoral counselor. AB - Discusses the development and popularity of short-term psychotherapy in relationship to the burgeoning field of managed health care. Views the role of pastoral counseling, pastoral counseling training, and the pastoral counselor in the context of the market economy of managed care. Claims that there is an incompatibility of pastoral counseling with managed behavioral health care, and calls for the return of pastoral counseling to the church. PMID- 10387599 TI - Why start from scratch? Starting with an empty office building, a New York company has created a modern nursing facility at a bargain price. PMID- 10387600 TI - Quest for smaller, homelike settings spurs suits. ADA used in court cases seeking alternatives to ICFs/MR. PMID- 10387601 TI - Long-term care plan gets rolling. PMID- 10387602 TI - Providing comfort to dying residents. PMID- 10387603 TI - Cultivating physician referral resources. PMID- 10387604 TI - Keeping time with fair labor standards. PMID- 10387605 TI - Assisted living funding poised for a streak. PMID- 10387606 TI - Caps leave providers with difficult choice. PMID- 10387607 TI - When the job market is tight ... ICFs/MR need creative recruiting strategies to be successful in finding and keeping staff. PMID- 10387609 TI - 1999 corporate profiles. PMID- 10387608 TI - A positive approach to resident concerns. PMID- 10387610 TI - Use and recognition of consensus standards in US premarket submissions. AB - In the autumn of 1997, the United States Center for Devices and Radiological Health published a draft guidance document on the use of IEC 60,601 standards in the evaluation of pre-market submissions for electromedical devices. One year later, an important legislative reform act caused this draft to be withdrawn because it allowed the Food and Drug Administration (FDA) to formally recognize standards covering all types of medical devices and not just those related to electrical products. This article discusses the benefits to manufacturers of this new FDA policy and the associated guidance documents that FDA has made available. PMID- 10387611 TI - Self-diagnostics and home monitoring: exploring new business opportunities. AB - The world is still shrinking. Advances in telecommunications are turning the concept of a global community into a reality. In the medical industry, this has translated into increased self-diagnostics and home monitoring. This article discusses a number of strategies for successful product positioning amidst technical revolution. PMID- 10387612 TI - Using lasers in medical device manufacturing. A technological evolution. AB - Laser technology has led to a number of valuable medical treatments, yet its greatest contributions are in the industrial and scientific sectors of health care. This article reviews a variety of applications where the laser tool has been successful in increasing the precision and reliability of industrial processes and medical instrumentation. Although many laser-based processes are replacements of conventional techniques, processes such as excimer photoablation offer new possibilities and are likely to define their own niches in the market place. PMID- 10387613 TI - Device-related nosocomial infection. Reducing infection with antimicrobial materials and coatings. AB - The occurrence of drug-resistant infections from hospital-based devices has soared in recent years. In response, industry has sought to develop a prophylactic approach with the materials used to construct devices. This article discusses one approach, a process that can be applied to a variety of materials and compares its performance with alternative methodologies. PMID- 10387614 TI - Know your intellectual property rights. AB - Dismissing intellectual property rights as someone else's responsibility may not be prudent. It can jeopardize a company's future business because any new product could be unprotected, or even worse, belong to someone else. This article explains how a little strategic planning and due diligence can help prevent unnecessary legal costs. Much of a company's value can be tied up in intangible assets such as patents, trademarks and copyright, and an intellectual property audit helps assess its true worth. PMID- 10387616 TI - Strength in diversity. PMID- 10387615 TI - A stain on the stainless? The role of steels in the new age. AB - Some recent events have suggested that the best days of steels have gone and that disposable plastics would be better for many applications. Although it cannot be denied that plastics have taken over many of the roles of steel during the last few decades and will, quite rightly, continue to do so in certain areas, this article argues the case for proper consideration for steels in other areas, where their properties are still of the greatest relevance. PMID- 10387617 TI - Market gains, but with some pain. PMID- 10387618 TI - Foresight begins with FMEA. Delivering accurate risk assessments. AB - If sufficient factors are taken into account and two- or three-stage analysis is employed, failure mode and effect analysis represents an excellent technique for delivering accurate risk assessments for products and processes, and for relating them to legal liability. This article describes a format that facilitates easy interpretation. PMID- 10387620 TI - Twenty-first century health care. AB - A dynamic, proactive health-care environment is beckoning. Fueled by consumer-led awareness, digital television, the Internet and a preoccupation with preventative health maintenance, it will define a new genre of products. In a series of provocative statements, this visionary article explores what the future may hold for diagnostics and medical devices. PMID- 10387621 TI - Which bar-code standard will best serve the medical device industry, EAN 128 or HIBC? PMID- 10387619 TI - An improved draft guidance to process validation. AB - The Global Harmonization Task Force has just released a new revised draft guidance on process validation. Although it contains much of the information included in the previous draft, some points have been added, and some that were confusing or not helpful have been clarified or eliminated. This article discusses the contents and usefulness of the latest draft process validation guidance document. PMID- 10387622 TI - Noxious chemical contamination risk. PMID- 10387623 TI - Materials for preformed rigid-tray packages. AB - This article assesses the materials used for medical device packaging that employs rigid preformed trays. It lists the basic requirements for these materials and summarizes the properties of each that make them useful for specific medical-packaging applications. A cost model illustrates how to properly select materials of similar properties on a cost-per-part basis. PMID- 10387624 TI - Medical packaging: more for less? AB - The market for medical devices is governed by the need to have validated products and processes, clean manufacturing environments and proven shelf-life. All this, often for products that are disposable, low-unit-cost items and, as such, highly price sensitive. This, in turn, has an impact on the businesses supplying the market. This article considers one such supply market, packaging, and the steps taken by packaging manufacturers to respond to these pressures while maintaining long-term, viable products. PMID- 10387625 TI - Packaging regulation update. Getting to grips with the unavoidable. PMID- 10387626 TI - A gym for the lungs. PMID- 10387627 TI - Novel screw caps for use in orthopaedic and trauma surgery. PMID- 10387628 TI - Fast-track development of a hand-held photometer. PMID- 10387629 TI - Small is beautiful: microparticle and nanoparticle technology in medical devices. AB - An increasing number of technologies require the production of ultra-small spherical particles, often described as micro-spheres or nano-spheres. They may be used for a variety of medical applications from cell encapsulation to serological diagnostic procedures. This article discusses some of the principles of this technology. PMID- 10387630 TI - Revisiting asbestos compliance. PMID- 10387631 TI - Safety metrics. PMID- 10387632 TI - The top 10 ways to achieve fund-raising success in the 21st century. PMID- 10387633 TI - Anatomy of a successful employee campaign. Securing the 21st century. PMID- 10387634 TI - The top three secrets of fund-raising success: communications, communications, communications! PMID- 10387635 TI - Relationship management--CEO involvement is key. PMID- 10387636 TI - The five delicate 'C's: keys to making the CEO-fund development relationship work. PMID- 10387637 TI - 1848-1998. Turning old into gold: anniversary marketing and corporate sponsorship opportunities. AB - The sesquicentennial was a unique opportunity to let the community know about the hospitals proud history. The hospital was featured throughout the year in articles and editorials in the local papers, professional hospital publications, and even nationally by Willard Scott on the "Today" show. The whirlwind of activities and events surrounding the anniversary created a spirit of pride in the hospital that will be invaluable for future fund-raising efforts. PMID- 10387638 TI - A sense of wonder. Raising money in the 21st century will be part of a greater task--teaching philanthropy. PMID- 10387639 TI - Practice brief. Managing health information relating to infection with the human immunodeficiency virus (HIV) (updated). American Health Information Management Association. PMID- 10387640 TI - Senate group keeps eye on Y2K. PMID- 10387641 TI - Honest mistake or fraud? Meeting the coding compliance challenge. PMID- 10387642 TI - Implementing information systems: a system development overview. PMID- 10387643 TI - Productivity: how do you measure up? PMID- 10387644 TI - Changes for the better: implementing four best practices. PMID- 10387645 TI - Navigating from data to excellence. PMID- 10387647 TI - Simple strategies for beating stress. PMID- 10387646 TI - A home health balancing act. PMID- 10387648 TI - OASIS (Outcomes and Assessment Information Set) cracks down on home care. PMID- 10387649 TI - Influence: making things happen. PMID- 10387650 TI - United we stand: professional alliances and networking. PMID- 10387651 TI - Alternative medicine: growing trend for the new millennium, Part II. PMID- 10387652 TI - Barriers to the adoption of computerized technology in health care systems. AB - The increased emphasis on national health care plans, cost reduction, and additional recordkeeping has given impetus to the adoption of computerized information technologies in hospitals. A series of case studies performed in large, multihospital health care systems revealed ten important barriers to the adoption of information technologies grouped as follows: knowledge problems, approval problems, design problems, and implementation problems. These aspects were uncovered by using focus studies and interviews with chief information officers, physicians, consultants, and medical staff and by consulting numerous journals in the field. The article describes the barriers that arise because of the special conditions in hospitals and shows how some institutions are working to eliminate these barriers. The strategic issues that should be studied to overcome these barriers are also discussed. PMID- 10387653 TI - Physician acceptance of telemedicine technology: an empirical investigation. AB - Fast-growing interest in telemedicine and increased investment in its enabling technology have made physician technology acceptance a growing concern for development and management of telemedicine. At the dawn of large-scale technology implementation by health care organizations around the globe, it is essential to understand physicians' attitudes toward use of telemedicine technology and their intention to use the technology. In this study, we used Theory of Planned Behavior to investigate technology acceptance among physicians who practiced in public tertiary hospitals in Hong Kong. Our data supported the investigated theory and the results suggest that attitude and perceived behavioral control are crucial to physician technology acceptance. Overall, physicians showed positive attitudes toward use of telemedicine technology and exhibited moderate intention to use the technology, primarily for clinical purposes. Implications for development and management of telemedicine also are discussed. PMID- 10387654 TI - Data and simulation modeling to determine the estimated number of primary care beds in Oklahoma. AB - Growth in managed care has resulted in an increased need for studies in health care planning. The article focuses on the estimated number of primary care beds needed in the state to provide quality services to all Oklahoma residents. Based on the 1996 population estimations for Oklahoma and its distribution to the 77 counties, a space-filling approach is used to determine 46 service areas, each of which will provide primary care service to its area population. Service areas are constructed such that residents of an area are within an acceptable distance to a primary care provider in the area. A simulation model is developed to determine the number of primary care beds needed in each service area. Statistical analysis on actual hospital data is used to determine the distributions of inpatient flow and length of stay. The simulation model is validated for acute care hospitals before application to the service areas. Sensitivity analysis on model input parameter values is performed to determine their effect on primary care bed calculations. The effect of age distribution on the bed requirement is also studied. The results of this study will assist the Oklahoma Health Care Authority in the development of sound health care policy decisions. PMID- 10387655 TI - Planning and managing computerized order entry: a case study of IT-enabled organizational transformation. AB - This article uses a model for technology-enabled organizational transformation to analyze a case study of computerized order entry at a medium-sized, urban hospital. The hospital achieved initial goals for direct order entry by physicians and improvements in patient care and is now using the system to implement disease management policies. The authors use the model to examine decisions and actions that facilitated or constrained effective implementation of the system and discuss the model's implications for managing implementation of computerized order entry technologies in health care systems. PMID- 10387656 TI - A framework for strategic information systems implementation in the United Kingdom health sector. AB - The implementation of information technologies in the United Kingdom health sector is a relatively recent phenomenon. Many of the developments have followed the patterns in the United States. One such example is that of Case Mix, introduced strategically as part of the Resource Management Initiative and aimed at the facilitation of both clinical and financial audit. Moreover, Case Mix was implemented alongside significant changes in hospital structure and culture, requiring clinicians to get involved in management tasks and decision making within the structure of the hospital, supported by a new information infrastructure. The success of such systems has varied significantly. A number of lessons can be learned from the way that the implementation was approached. This article stems from a research project focusing longitudinally on the implementation of Case Mix in four UK hospitals. It draws a number of findings from the cases, and importantly, explicates a framework for strategic information systems implementation, as generated from the cases and supported by the extant literature. Such a framework has implications for both theory and practice, and assists in the understanding of what is often a dynamic and poorly understood situation. PMID- 10387657 TI - Matching system requirements to organizational function. AB - The Organizational Behavior discipline suggests that there are various functions of management within organizations. The functions require that people make decisions as part of the ordinary operations of the organization. These decisions are dependent on timely, accurate, and reliable information. Because the functions of management vary, the information requirements to support those functions also vary. An information system that is designed on the basis of these varying information needs for different parts of the management structure thereby producing an information system that is integrated rather than fragmented, promotes the probability of better decisions and thus enhances the organization's competitive position in its environment. PMID- 10387658 TI - Strategic relevance and accountability expectations: new perspectives for health care information technology design. AB - In this article, we discuss the traditional systems analysis perspective on end user information requirements analysis and extend it to merge with the new accountability expectations perspective to guide the future planning and design of health organization information systems. Underlying the strategic relevance of health care information technology (HCIT) are three critical questions: (1) What is the ideal HCIT model for the health organization in terms of achieving strategic expertise and competitive advantage? Specifically, how does this model link industry performance standards with organizational performance and accountability expectations? (2) How should the limitations of past HCIT models be reconciled to the benefits presented by the superior arrangement of the ideal model in the context of changing accountability expectations? (3) How should alternative HCIT solutions be evaluated in light of evidence-based accountability and organizational performance benchmarking? Insights into these questions will ensure that health care managers, HCIT practitioners and researchers can continue to focus on the most critical issues in harnessing today's fast-paced changing technologies for evolving strategically relevant, performance-based health organization systems. PMID- 10387659 TI - The hospitalist trend keeps on growing but physician resistance, though diminished, remains a challenge. PMID- 10387660 TI - Quality initiative combs the nation for innovative practices. PMID- 10387661 TI - Who's a hospitalist--and why? PMID- 10387662 TI - A new model emerges as primary care physicians and mental health professionals collaborate. PMID- 10387663 TI - Process waste mapping: a pain-free way to get rid of the "CRUD" in your organization. PMID- 10387664 TI - The power of the purse. More and more, it's women who control the charity. PMID- 10387665 TI - The grief brigade. PMID- 10387666 TI - Beyond depression. What do those "mood drugs" really do? PMID- 10387667 TI - The new mister natural. PMID- 10387668 TI - Homeopathic e-mail. PMID- 10387669 TI - Thrombolytic therapy for acute ischemic stroke. PMID- 10387671 TI - Taxing away the uninsured. PMID- 10387670 TI - Business principles for air medical professionals. Part 5: Choosing a consultant. PMID- 10387672 TI - Managed care profitability on the way up. PMID- 10387673 TI - What have they done for you lately? PMID- 10387674 TI - Tackling costs one disease at a time. PMID- 10387675 TI - The trouble with medical innovation. PMID- 10387676 TI - Weighing in on LTC insurance. PMID- 10387677 TI - Who's minding the PPOs? PMID- 10387678 TI - Data watch. Pressure on partnering. PMID- 10387679 TI - Rise & fall of MedPartners. PMID- 10387680 TI - Specialist collaboration strategies--roundtable discussion. PMID- 10387681 TI - A rally call to universal health care management. PMID- 10387682 TI - Australia's health system: the search for a cure. The Australian Democrats' discussion paper on Australia's health system. AB - The Australian Democrats' believe that Australia is currently at a critical time in the development of its health care system. The twin pressures of higher demands and inadequate funding have led to alarmingly heavier pressure on the public health infrastructure. Increasingly, the choices confronting society is one of how to allocate limited resources in the health sector. Awareness is growing on the impact wider issues such as unemployment, fragmentation of communities are having on society's health. In the face of new challenges confronting Australia's health system of the needs of an aging population, adoption of new developments in medical treatment and technology and new relationships which are developing between the public and private health sector, the Democrats acknowledge that although there are no easy responses, it is important, nonetheless to recognise their complexities and work collaboratively towards viable solutions. Through this discussion paper, based on consultation with a wide range of stakeholder groups and individuals, the Australian Democrats' have sought to outline the options for addressing the challenges and debate which will serve to contribute to the development of a fair, efficient and equitable health care system for all Australians, now and into the future. PMID- 10387683 TI - The Internet and poverty: real help or real hype? Panos Briefing No. 28. AB - Governments, donors and development organisations are rushing to realise the benefits that Internet access promises in the fight against poverty. But are the benefits it has brought so far merely isolated examples or are they signs that a revolution is underway? Access to information is an essential condition of development. The Internet has prompted a change in development thinking and many donor and multilateral lending organisations are radically reshaping their policies for the new information age. But is the enthusiasm among donors for spending on Internet development diverting funds from more traditional forms of development assistance? In terms of its most adaptive component, the World Wide Web, the Internet is still only four years old. Real hype or real help? The jury is still out. Quite simply, it is still too soon to tell. PMID- 10387684 TI - Intelligent buildings: a self regulating environment. AB - Developments in technology mean that "intelligent buildings" are now fact, not fiction. There is a fine line that must be followed when developing an intelligent building. The occupier must understand what is being provided and what is expected from the completed building. Careful observation of the proposed occupiers operating regime is a must. The most elegant solution to designing an intelligent building lies in the design of its fabric, so called passive engineering. Passive engineering emulates what happens in the natural world: a true intelligent building will provide a self-regulating internal environment by purely natural means. PMID- 10387685 TI - Speech to the Fifty-First World Health Assembly--Geneva, 13 May 1998. PMID- 10387686 TI - The challenges in a volatile marketplace. PMID- 10387688 TI - Help your patients light up ... their lives. PMID- 10387689 TI - Physician supply and demand in the new millennium. PMID- 10387690 TI - How to navigate employee relations. PMID- 10387687 TI - Giving back to the community. Interview by Jane Byrne. PMID- 10387691 TI - A warning for hospitals. A challenge for hospital trustees. PMID- 10387692 TI - Adding some "cheer" to the ER. PMID- 10387693 TI - Overcoming hurdles. AB - Human resource professionals in Michigan face significant hurdles in balancing the rights of employees under federal and state employment laws. Perhaps none cause more difficulties than the Americans with Disabilities Act (ADA), the Family Medical Leave Act (FMLA), and Michigan's Workers' Disability Compensation Act. Employees frequently have conflicting rights under these statutes so that you cannot assume that since you have complied with one set of laws, you have complied with the others. While these three laws appear to address different employment-related concerns, difficulties arise when employers attempt to juggle them. PMID- 10387694 TI - Partners in change. PMID- 10387695 TI - Managing the uncompensated care problem, Part III. AB - Six Community Initiatives of a Comprehensive, Uncompensated Care Program. 1. expanded hospitalization coverage. 2. managed seamless continuum of care. 3. humane, disciplined collection of bad debts. 4. advocacy for potential recipients of uncompensated care. 5. philanthropic support. 6. increased community focus on improved health status. PMID- 10387696 TI - Employees invest and everyone profits. PMID- 10387697 TI - E-mail's from hell: don't get burned. AB - E-mail's growing popularity and unmonitored use is putting employers at risk. E mail messages are not the personal property of the employee who generates them, and therefore, workers have no right to privacy regarding them. The National Institute of Business Management recommends that employers monitor employees' e mail messages for evidence of discriminatory material. PMID- 10387699 TI - The human resources journey. PMID- 10387698 TI - Don't volunteer for trouble. AB - Volunteers have always played an important role in the continuum of care a hospital provides to its patients and the community, and the reliance on hospital volunteers appears to be on the rise. This may be one of many responses to widespread cost containment efforts due primarily to an increase in reimbursement pressures. The use of volunteers raises unique liability issues, which warrant periodic review within the overall risk management plan of a hospital. A recent Michigan case has brought renewed attention to the use of volunteers. PMID- 10387700 TI - Managing data, managing care. How Aetna U.S. Healthcare is using a massive data warehouse to better manage health care. PMID- 10387701 TI - Are these cards a smart move? PMID- 10387702 TI - Records on the Net: an update. PMID- 10387703 TI - A man on a mission. Interview by John McCormack. PMID- 10387704 TI - Why opt for thin clients? A nursing home selects the technology to control its long-term maintenance costs. PMID- 10387705 TI - Winning support for physician order entry. PMID- 10387706 TI - Hospitals weigh a growing number of hardware options. Choosing the right devices can determine the success of inpatient point-of-care computing initiatives. PMID- 10387707 TI - The patient as the record builder. PMID- 10387708 TI - But are they satisfied? Technology enables organizations to get faster and more accurate results from patient satisfaction surveys. PMID- 10387709 TI - Telemedicine providers' progress impeded at the border. The need for physicians to obtain a medical license in every state in which they practice slows the growth of telemedicine. PMID- 10387710 TI - Integrating physicians in the "post-PPM" era: ten innovative models for managing physician organizations in the 21st century. PMID- 10387711 TI - Group practice is the future of medicine, but who will own and manage the groups? PMID- 10387712 TI - Innovative models: ten options for managing physician organizations. PMID- 10387713 TI - Pinching pennies in the ED: impress administrators with creative solutions. PMID- 10387714 TI - Patient care, efficiency helps you cut costs. PMID- 10387715 TI - Get an edge by giving MCOs a report card. PMID- 10387716 TI - Patient transport codes give paramedics options. PMID- 10387717 TI - Contact capitation: boon or bane? AB - Capitation has shown itself to be a challenging form of payment to work with from a number of standpoints. Not only does it require the alignment of relationships between and among primary care physicians and specialists via explicit contracting and financial terms, but it requires organization of all types of physicians across entire health-care markets, and sophisticated data analysis for success. Now, a still-new form of capitation called "contact capitation" is evolving. It seems to hold promise in terms of resolving some short-term administrative problems. But can it offer improvement in alignment of incentives long-term? This article looks at the growth of contact capitation as a payment distribution strategy, and its opportunities, pitfalls, and issues in terms of physician organization and integrated delivery objectives. PMID- 10387718 TI - Pacific Business Group on Health spotlights MD group quality. AB - For the first time ever, a powerful California-based purchasing coalition goes directly to the provider level to spotlight quality achievement, hoping to stimulate broader purchaser and consumer interest in physician group quality measurement. PMID- 10387719 TI - Managed care and charity care: on a collision course? PMID- 10387720 TI - HCFA guidance places limitations on distinct parts, new provider certification. PMID- 10387721 TI - Discounted ancillary contracts could violate anti-kickback law, says OIG. PMID- 10387722 TI - Seventh circuit appeals court rules that LPNs are not supervisors. PMID- 10387723 TI - Fiscal 1998 annual financial results of post acute/subacute companies. PMID- 10387724 TI - Intermediaries to begin random medical reviews of SNF PPS bills; memo revisits RUGs deeming issue. PMID- 10387725 TI - Industry warns of financial crisis during HCFA town hall meeting, urges immediate changes to PPS. PMID- 10387726 TI - Supreme Court agrees to rule on challenge to Medicare regulations. Illinois Council on Long-Term Care v. Shalala. PMID- 10387727 TI - How to avoid legal troubles with your facility's staff. PMID- 10387728 TI - An attitude adjustment: handling difficult employees. PMID- 10387729 TI - It's show time! How to plan and execute successful culinary demonstrations. PMID- 10387730 TI - Problem-based learning: a method that encourages critical thinking. PMID- 10387731 TI - Applying the principles of problem-based learning: two examples. PMID- 10387732 TI - More tips from the trayline doctor. Part 2: How to ensure HACCP (Hazard Analysis and Critical Control Point) compliance. PMID- 10387733 TI - Helping your staff become more receptive to change. PMID- 10387734 TI - Calculating the true cost of employee turnover. PMID- 10387735 TI - The Supreme Court opens the door wide for harassment claims. PMID- 10387736 TI - Blending a diverse health care team into a rose garden. PMID- 10387738 TI - Self-managed work teams. What are they and how do they work? PMID- 10387737 TI - Developing admission agreements. Market your assisted living community and avoid litigation. PMID- 10387739 TI - Prepare now for Medicare full encounter data reporting. PMID- 10387740 TI - Accurate encounter data is effective clinical tool to provide better patient care. AB - Consider using encounter data for more than risk-adjusted Medicare or Medicaid payment. Providers with a New Jersey-based Medicaid plan and with a Medicare risk organization in Media, PA, expect to use encounter information to profile physicians and improve processes of care. Find out what they're planning. PMID- 10387741 TI - Use these 5 tips to mitigate exposure when assuming risk for Medicare pharmacy. AB - Should providers take on Medicare pharmacy risk in today's environment? With drug costs shooting out of sight, experts say no. But what if you're already stuck? Mitigate your risk with these five tips. PMID- 10387742 TI - More providers taking risk for Medicaid behavioral health. AB - Providers making inroads into managed Medicaid behavioral health. A new study shows that traditional public agencies and providers, not commercial managed care organizations, are running behavioral health programs in managed Medicaid. Even Medicaid HMOs with behavioral health benefits integrated in their program are turning behavioral health over to providers, putting them at full risk. See how one plan's behavioral health contractor is rising to the challenge. PMID- 10387743 TI - State rules, changing local markets create chance for PSOs to take over Medicaid risk. AB - Data File: Are provider organizations taking over Medicaid risk? The answer depends on the market, as demonstrated in these three analyses of enrollment trends in New York, Florida, and Ohio. PMID- 10387744 TI - Look beyond the sales pitch when evaluating capitation IS. PMID- 10387745 TI - How much data do you need to predict variability? Maybe less than you think. AB - How much data do you need to predict variability in managing risk? By using run charting and control charting, your cycle time needed to predict utilization and cost may be much shorter than commonly believed necessary. PMID- 10387746 TI - Higher profitability, lower operating costs are emblems of better performing practices. PMID- 10387747 TI - Physician compensation package ramps up to meet marketplace changes. AB - A small multispecialty group had to retool its compensation system "practically overnight" when the state unveiled a managed Medicaid plan and capitation began to account for nearly half its revenues. Here's how they did it while enhancing relationships with specialists and PCPs alike. PMID- 10387748 TI - Despite growing popularity, pain management not yet ready for capitation. PMID- 10387749 TI - Perspectives. Indefinite results in ABMT (autologous bone marrow transplantation) trials add to challenges for practice standards, quality assurance in cancer care. PMID- 10387750 TI - Perspectives. Policymakers grapple with foundations of process for coverage decisions, appeals. PMID- 10387751 TI - Marketplace. GM applies auto plant techniques to help hospitals improve efficiency. PMID- 10387752 TI - Perspectives. Crisis or crying game? SNFs protest BBA cuts, but others warn that new PPS might still result in overpayment. PMID- 10387753 TI - Perspectives. Reports from the field suggest some quality tools work best at local level. PMID- 10387754 TI - Partners in HF (heart failure) care: the new paradigm. PMID- 10387755 TI - Optimizing the inpatient management of diabetes. PMID- 10387756 TI - Doctors unions, in accelerating trend, refigure the managed care equation. PMID- 10387757 TI - Harnessing the vitality and power of intimacy. PMID- 10387758 TI - Gift averaging: don't even think about it! PMID- 10387759 TI - Do it right: selecting the right architect and negotiating the right relationship. PMID- 10387760 TI - Preparing for organizational change. PMID- 10387761 TI - Getting past first base with your new donor: three rules for getting a second date. PMID- 10387762 TI - Your newsletters: are they worth their weight in gold? PMID- 10387763 TI - Wanted: a new breed of physician drivers for healthcare's nitroglycerin trucks. PMID- 10387764 TI - Challenges in developing physician leadership and management. AB - Many of the issues confronting healthcare organizations require physician involvement and understanding, and the physician executive is a tool to achieving physician participation. Physician leaders can become the mediators between physicians and organizational management, minimizing miscommunication and maximizing agreement and understanding. Yet few doctors seem willing to stand up and speak positively for the plans and proposals that will move the institution forward, and healthcare executives are often frustrated by physician leadership that fails to articulate and implement the vision and objectives of the organization. Understanding physician leadership and exploring the challenges in managing and leading physicians require an understanding of the physician mindset -a completely different mindset than that of the typical healthcare executive. Beginning with a discussion of the unique situation faced by physicians in leadership positions, this article attempts to define the obstacles faced by both the physician and the executive in developing the role of physician executive. After reviewing the opportunities open to physician executives for improving leadership ability, the author presents the essential characteristics and core skills for effective leadership. The second half of the article suggests ways in which an organization can reemphasize physician leadership development within an organization from selection of potential candidates to creating training and networking opportunities and offering appropriate incentives. PMID- 10387765 TI - Transitional leaders for transitional times. PMID- 10387766 TI - Developing physician leaders in today's hospitals. PMID- 10387767 TI - What's your strategy for managing knowledge? AB - The rise of the computer and the increasing importance of intellectual assets have compelled executives to examine the knowledge underlying their businesses and how it is used. Because knowledge management as a conscious practice is so young, however, executives have lacked models to use as guides. To help fill that gap, the authors recently studied knowledge management practices at management consulting firms, health care providers, and computer manufacturers. They found two very different knowledge management strategies in place. In companies that sell relatively standardized products that fill common needs, knowledge is carefully codified and stored in databases, where it can be accessed and used- over and over again--by anyone in the organization. The authors call this the codification strategy. In companies that provide highly customized solutions to unique problems, knowledge is shared mainly through person-to-person contacts; the chief purpose of computers is to help people communicate. They call this the personalization strategy. A company's choice of knowledge management strategy is not arbitrary--it must be driven by the company's competitive strategy. Emphasizing the wrong approach or trying to pursue both can quickly undermine a business. The authors warn that knowledge management should not be isolated in a functional department like HR or IT. They emphasize that the benefits are greatest--to both the company and its customers--when a CEO and other general managers actively choose one of the approaches as a primary strategy. PMID- 10387768 TI - Competing with giants. Survival strategies for local companies in emerging markets. AB - The arrival of a multinational corporation often looks like a death sentence to local companies in an emerging market. After all, how can they compete in the face of the vast financial and technological resources, the seasoned management, and the powerful brands of, say, a Compaq or a Johnson & Johnson? But local companies often have more options than they might think, say the authors. Those options vary, depending on the strength of globalization pressures in an industry and the nature of a company's competitive assets. In the worst case, when globalization pressures are strong and a company has no competitive assets that it can transfer to other countries, it needs to retreat to a locally oriented link within the value chain. But if globalization pressures are weak, the company may be able to defend its market share by leveraging the advantages it enjoys in its home market. Many companies in emerging markets have assets that can work well in other countries. Those that operate in industries where the pressures to globalize are weak may be able to extend their success to a limited number of other markets that are similar to their home base. And those operating in global markets may be able to contend head-on with multinational rivals. By better understanding the relationship between their company's assets and the industry they operate in, executives from emerging markets can gain a clearer picture of the options they really have when multinationals come to stay. PMID- 10387769 TI - Unbundling the corporation. AB - No matter how monolithic they may seem, most companies are really engaged in three kinds of businesses. One business attracts customers. Another develops products. The third oversees operations. Although organizationally intertwined, these businesses have conflicting characteristics. It takes a big investment to find and develop a relationship with a customer, so profitability hinges on achieving economies of scope. But speed, not scope, drives the economics of product innovation. And the high fixed costs of capital-intensive infrastructure businesses require economies of scale. Scope, speed, and scale can't be optimized simultaneously, so trade-offs have to be made when the three businesses are bundled into one corporation. Historically, they have been bundled because the interaction costs--the friction--incurred by separating them were too high. But we are on the verge of a worldwide reduction in interaction costs, the authors contend, as electronic networks drive down the costs of communicating and of exchanging data. Activities that companies have always believed were central to their businesses will suddenly be offered by new, specialized competitors that won't have to make trade-offs. Ultimately, the authors predict, traditional businesses will unbundle and then rebundle into large infrastructure and customer relationship businesses and small, nimble product innovation companies. And executives in many industries will be forced to ask the most basic question about their companies: What business are we really in? Their answer will determine their fate in an increasingly frictionless economy. PMID- 10387771 TI - Tailored, not benchmarked. A fresh look at corporate planning. AB - In today's competitive markets, every company has an action plan. Yet for most managers, the processes used to create these plans don't work. The root of the problem, suggests Campbell, may be that too many companies benchmark their processes and by doing so, prevent managers from focusing on what is unique to their situation. Good planning processes, the author argues, are not generic processes but ones in which both analytic techniques and organizational processes are carefully tailored to the needs of individual businesses and to the skills of corporate managers. The author cites examples of three companies that have successfully individualized their processes: Granada, Dow Chemical Company, and Emerson Electric. A mature electrical-products business such as Emerson, he says, has different planning needs than a fast-growing entertainment business like Granada or a highly cyclical chemicals business like Dow. Different chief executives may have different insights about how to go about adding value. Take the CEOs of Granada and Dow. Both set tough targets to stretch their businesses, but the way each CEO gets his managers to commit to his targets differs considerably. Bad planning can actively destroy value, the author says. It wastes people's time and money. It sends the wrong signals to managers. It can even lead managers to follow bad advice. That's why managers should go to the effort of reexamining and possibly changing their company's planning process. PMID- 10387770 TI - Web site blues. AB - So far, Rachel Soltanoff's instincts had been right. As CEO in this fictional case study, she had successfully navigated TradeRite Software's transition from a news service for stockbrokers to a $70 million provider of shrink-wrapped software geared toward both brokers and the growing day-trader market. Now a well financed start-up, Stock-net.com, was testing a very competitive product that traders could download directly over the Web. And TradeRite's Web site was nothing more than a collection of elaborate marketing brochures. Rachel knew she needed to start selling over the Web. But the e-commerce consultants she had hired to set up her Web store were behind schedule, and their 21-year-old CEO had just resigned. Her product manager, Lisa Bandini, was working overtime to transform TradeRite's entire product line into Web-aware applications to match Stocknet's, and Rachel had $2.5 million to launch them. But the consultants said it would take $5 million just to rent e-commerce capabilities. Ace sales VP Brian Rockart thought the company had already wasted too much time and money--money from his budget--on its Web site. Marketing VP Rob Collins thought TradeRite should focus on its core stockbroker customers. Chief Technical Officer Joe Martinez doesn't want to go ahead without a pilot project. Should Rachel try to convince Brian, Rob, and the rest of the senior management team that e-commerce is the way to go? Four commentators offer advice. PMID- 10387773 TI - Driving change: an interview with Ford Motor Company's Jacques Nasser. Interview by Suzy Wetlaufer. AB - What happens when the world is changing but your organization isn't? And what if that organization has 340,000 employees in 200 countries? In this interview, Jacques Nasser, the new CEO of Ford Motor Company, talks with HBR editor Suzy Wetlaufer about these challenges and explains how his company is overcoming them through a unique education program. Since its very beginnings, says Nasser, Ford has comprised dozens of far-flung divisions and units, each with its own "fiefdom" mind-set. The fiefdoms didn't share information, let alone great ideas. Such behavior stifled creativity and drove up costs. Today's global environment demands a new and different way of doing business, says Nasser, and to that end, Ford has launched a multifaceted teaching initiative that will reach every one of Ford's employees by year-end. The goal of the program: to help employees view the company in its entirety as shareholders do, and then act that way too. At the heart of the initiative is the teachable point of view, a five-part written explanation of what a person knows and believes about what it takes to succeed in business. It is more than just a document to be discussed and then filed. It has proven to be a powerful tool for organizational transformation, and not only at Ford. In a commentary accompanying Nasser's interview, Noel Tichy, leadership expert and consultant to Ford describes the building blocks of the teachable point of view and explores how it can be implemented in any organization determined to change for the better. PMID- 10387772 TI - Managing oneself. AB - Throughout history, people had little need to manage their careers--they were born into their station in life or, in the recent past, they relied on their companies to chart their career paths. But times have drastically changed. Today, we must all learn to manage ourselves. What does that mean? According to Peter Drucker, it means we have to learn to develop ourselves. We have to place ourselves where we can make the greatest contribution to our organizations and communities. And we have to stay mentally alert and engaged during a 50-year working life, which means knowing how and when to change the work that we do. It may seem obvious that people achieve results by doing what they are good at and by working in ways that fit their abilities. But, Drucker says, very few people actually know--let alone take advantage of--their unique strengths. He challenges each of us to ask ourselves fundamental questions: What are my strengths? How do I perform? What are my values? Where do I belong? What should my contribution be? Don't try to change yourself, cautions Drucker. Instead, concentrate on improving the skills you have and accepting assignments that are tailored to your individual way of working. If you do that, you can transform yourself from an ordinary worker into an outstanding performer. Successful careers today are not planned out in advance. They develop when people are prepared for opportunities because they have asked themselves those questions, and they have rigorously assessed their unique characteristics. This article challenges readers to take responsibility for managing their futures, both in and out of the office. PMID- 10387774 TI - New thinking on how to link executive pay with performance. AB - As the stock market began its ascent in the mid-1990s, executive pay--always the subject of heated debate--mounted along with it. That's because among the largest U.S. companies, stock options now account for more than half of total CEO compensation and about 30% of senior operating managers' pay. One problem became particularly clear during the bull market's astonishing run: even below-average performers reap huge gains from stock options when the market is rising rapidly. The author proposes steps to close the gap between existing compensation practices and those needed to promote higher levels of achievement at all levels of the corporation. For top managers, he recommends replacing conventional stock options with options that are tied to a market or peer index. Below-average performers would not be rewarded under such plans; superior performers could, depending on the way plans were structured, receive even more. He notes that managers at the business unit level should not be judged on the company's stock price--over which they have little control--and advocates an approach that accurately measures the value added by each unit. Finally, he suggests how certain indicators of value can be used to measure the contribution of frontline managers and employees. The concept of pay for performance has gained wide acceptance, but the link between incentive pay and superior performance is still too weak. Reforms must be adopted at all levels of the organization. Shareholders will applaud changes in pay schemes that motivate companies to deliver more value. PMID- 10387775 TI - Quick and continuous improvement through kaizen blitz. AB - It is our objective to provide you with a step-by-step approach to conducting a kaizen blitz within two days and describe how to achieve dramatic performance improvement with employee buy-in through this process. Kaizen blitz has been used dozens of times by the authors, and in some instances the same area has been blitzed as many as four times, with significant improvements each and every time. Employees have even taken it on themselves to conduct informal blitzes as a continuing improvement effort after a formal blitz has been conducted in their area. Blitzes can succeed in a variety of environments. The morning after the employees of one company attended this presentation, they self initiated a mini blitz and discovered opportunities for improvement that they enthusiastically presented to management. PMID- 10387776 TI - Rapid-fire improvement with short-cycle kaizen. AB - Continuous improvement is an attractive idea, but it is typically more myth than reality. SCK is no myth. It delivers dramatic improvements in traditional measures quickly. SCK accomplishes this via kaizens: rapid, repeated, time compressed changes for the better in bite-sized chunks of the business. PMID- 10387777 TI - Case study on the 5S program: the five pillars of the visual workplace. AB - This case study will explain the 5S concepts and how The Clarkson Company has begun implementing such a program, and how it is a foundation for continuous improvement. PMID- 10387778 TI - Baseline budgeting for continuous improvement. AB - This article is designed to introduce the techniques used to convert traditionally maintained department budgets to baseline budgets. This entails identifying key activities, evaluating for value-added, and implementing continuous improvement opportunities. Baseline Budgeting for Continuous Improvement was created as a result of a newly named company president's request to implement zero-based budgeting. The president was frustrated with the mind-set of the organization, namely, "Next year's budget should be 10 to 15 percent more than this year's spending." Zero-based budgeting was not the answer, but combining the principles of activity-based costing and the Just-in-Time philosophy of eliminating waste and continuous improvement did provide a solution to the problem. PMID- 10387779 TI - Creative problem-solving: an approach to generating ideas. AB - An excellent starting point for exercising creativity is the area of problem solving. With a bag of creative problem solving tools and techniques, problems will no longer represent setbacks but instead, opportunities to introduce innovations that will support the company's initiative of continuous improvement. PMID- 10387780 TI - Transforming your manufacturing organization into a learning organization. AB - Many experts believe the only sustainable advantage an organization will have in the future is its ability to learn faster than its competitors. This competitive advantage can be achieved by transforming the organization into a learning organization. This article describes the basic elements of a learning organization and how to transform an organization to focus on learning. The article also describes methods for evaluating the training and learning that occurs within the organization to ensure that training dollars are wisely spent. Finally, the article describes what individuals can do to focus on their own learning and personal development within an organization. PMID- 10387781 TI - Testing a model of long-term alliance success. AB - The remarkable consistency of buyer and supplier perspectives was the most striking feature of this research. Although significant differences did exist, the dominant perspective illustrated a strong cohesion between partners. AS such, it would appear that these alliances do represent "best in class" relationships and thus may provide valuable insight for academics and practitioners into what makes alliances succeed--or fail. PMID- 10387782 TI - Proactive performance measures. AB - Adoption of a process of careful selection and implementation generating the appropriate proactive (leading) performance measures will provide an important tool with which to progress. When used in combination with traditional financial performance measures, proactive performance measures contribute to our cache of competitive weapons. PMID- 10387783 TI - Breaking through barriers to successful empowerment. AB - In today's ever-changing environment, most companies are undergoing some kind of transformation, whether it is implementing new information systems, reengineering manufacturing methods, rightsizing, tackling paradigms, or simply dealing with demand changes. This transformation often is difficult based on old ways of managing the most important corporate asset-people. Many companies have initiated the processes of team building and people empowerment, but few are able to permeate an entire organization with these concepts. This presentation will provide the successful methods that certain organizations are using to create empowered individuals and teams, and discuss proven techniques for implementing these ideas and ensuring success. PMID- 10387784 TI - The "special K" in kaizen. AB - Do you want a strategy that guarantees the success of kaizen in your organization? This article will provide insights as to why one organization may succeed and another fails. Learn why the "Special K" is the difference. Idea will be presented on how to develop the "Special K" in others and ourselves. You will be challenged to become a "kaizener". PMID- 10387785 TI - Is anybody listening? PMID- 10387786 TI - Sharp interpersonal skills: your key to business success. AB - In today's more participative work environments, it is more important than ever to have strong interpersonal skills. Several recent studies cite interpersonal skills as a critical element in the selection of leader's in today's organizations. No longer are we relying upon power and control, but rather on empowerment and commitment. This article deals with building interpersonal working relationships, the type that helps to create synergy and teamwork within a workgroup or organization. PMID- 10387787 TI - Business writing: a foolproof system for getting started. AB - Writing well is a valuable skill, but one often lacking among higher and lower level employees. Most people recognize good writing when they see it, but are at a loss to define exactly what it is. I define it as writing that's easy to follow and understand. PMID- 10387789 TI - Radio frequency solutions to shorten cycle time and increase accuracy. AB - As a result of the continuous improvement using RF bar code technology, significant gains in inventory accuracy and cycle time reductions have been achieved. PMID- 10387788 TI - What? Me speak in public: what if somebody sees me? AB - The purpose of this paper is to highlight the specific critical areas of public speaking. It deals with the important and often forgotten areas of the presentation: the objective statement and lesson plan, key platform skills, and the motivational environment. PMID- 10387790 TI - Telephone services. A year down the line. PMID- 10387791 TI - Primary care groups. Alter ego. PMID- 10387792 TI - Primary care groups. Chemistry lessons. PMID- 10387793 TI - Never mind the quality... PMID- 10387794 TI - NHS reorganisation. A rose-tinted spectacle. AB - The 1974 reorganisation of the NHS was the most radical to date. It abolished the involvement of local authorities in health, set up community health councils, introduced area health authorities and changed the management of family doctor services. The changes increased the power of hospitals. PMID- 10387795 TI - The book of revelations. PMID- 10387796 TI - Mental health services. Poor relations. AB - The case for London requiring greater resources for mental health services than other parts of the country has not been proved. Liverpool, Birmingham and Manchester are among the six most deprived areas in England. Spending per capita on mental health services in inner London is double that in Birmingham and Liverpool and 40 per cent higher than in Manchester. A national strategy is needed to address inequities in funding. PMID- 10387797 TI - Underperforming doctors. Back from the brink. AB - The UK has a poor record on retraining underperforming doctors and rehabilitating sick ones. But that is set to change. Wendy Moore examines new attitudes and plans of action aimed at meeting the demands of the clinical governance revolution. PMID- 10387798 TI - Underperforming doctors. Recovery services. AB - In the US, state medical licensing boards have the power to prevent sick doctors from practising if they do not undergo treatment. Every state has a programme for impaired doctors, most including a two-week hospital stay involving intensive psychotherapy. Research on recovery is limited, but some studies have found recovery rates of 75 per cent. PMID- 10387799 TI - Finance managers--figure heads. Interview by Ann Dix. PMID- 10387800 TI - Genetics services. Inheritance tacks. PMID- 10387801 TI - Genetics services. Briefing encounters. AB - Developments in genetic science have huge implications for NHS services. Demand for genetic testing is set to increase. The Public Health Genetics Unit aims to link academic research, policy making and clinical practice, and stimulate debate about the provision of genetic services. PMID- 10387802 TI - Pharmaceuticals. Plunder and pillage. AB - The government has proposed a statutory scheme to regulate prices and profits on the sale of drugs to the NHS. Drug companies which don't cooperate could be liable to heavy fines, but the government has failed to spell out the precise conditions under which such penalties would apply. Defining what comprises excessive prices or profits is likely to present a number of empirical problems. Experience overseas suggests that the scheme will fail to halt the inexorable rise in the NHS drugs bill. PMID- 10387803 TI - You ain't seen nothing yet. PMID- 10387804 TI - Primary care trusts. States of flux. AB - Primary care trusts are similar to US health maintenance organisations in that they act as both commissioners and providers of services. There are already innovative primary care organisations in the NHS and the private sector that can be classed as embryonic PCTs. Experience of these organisations, assessed in relation to HMOs, can inform the development of PCTs. PMID- 10387805 TI - Primary care managers. New men. Interview by Jeremy Davies. PMID- 10387806 TI - Data briefing. Health risks. PMID- 10387807 TI - Private clinics for employees as a Dutch solution for waiting lists: economic and legal arguments. AB - Private clinics for employees have emerged on a small scale during the last few years in Dutch health care. They represent new possibilities for ill employees who have to wait for medical treatments on waiting lists. Earlier proposals to allow employers in close co-operation with health insurers to start initiatives for special clinics for employees have been confronted with all kind of arguments which were considered to form serious obstacles, from theoretical, legal and economic perspectives. The European Court of Justice plays a decisive role in deciding whether free establishment and access is approved according to the rules of the EC Treaty. Private clinics can be legally allowed in the interests of the common good. However, priority treatment of a person can only be justified if the intended purpose is justified and if the priority treatment is suited as a means to (partly) realise this purpose. Furthermore, possible negative consequences may not be unreasonable in the light of the intended consequences. In this article it will be argued that both from an economic and from a legal perspective, based on national and European law, the introduction of special clinics for employees could be allowed. The main argument is that they could be introduced on a just and equitable basis. PMID- 10387808 TI - Are market-oriented health insurance reforms possible in Latin America? The cases of Argentina, Chile and Colombia. AB - The process of health care reform benefits tremendously from comparing characteristics and performance across nations. This paper studies market oriented health insurance reforms in three Latin American countries: Argentina, Chile and Colombia. Chile allowed private health insurers to compete for workers payroll contributions in the 1980s, permitting the modernization of the private health sector but relatively impoverishing the public health sector as a consequence of selection practices by private carriers. In the 1990s, Argentina and Colombia started liberalizing the health insurance sector but using policies to avoid the adverse effects encountered in the Chilean experience. These policies are scrutinized while challenges for these and future health insurance reform processes are discussed. PMID- 10387809 TI - A framework for incorporating cost-effectiveness in evidence-based clinical practice guidelines. AB - In England, recent health care reforms emphasise the role of clinical guidelines in promoting effective and efficient health care. Introducing economic data into guidelines raises some methodological issues: specifically, the provision of valid and generalisable cost estimates, the weight placed upon cost 'evidence', and the presentation of cost-effectiveness information in a manner accessible to clinicians. A series of primary care guidelines, explicitly including consideration of health economic information, have recently been published, intended to help clinicians to aggregate the attributes of treatment choices to derive treatment recommendations consistent with both the clinical decision making process and social objectives. Clinicians involved in developing guidelines responded well to the process and consistently managed to agree treatment recommendations, often after considerable debate about the evidence for treatment. In none of the guideline areas, all of which addressed common diseases, was there adequate information to estimate a cost per quality-adjusted life-year, and it is unclear how helpful this approach would have been had it been possible. The implications of this method are discussed, guidance offered for economists new to guideline development and future areas of work identified. PMID- 10387810 TI - Constraints on the retreat from a welfare-orientated approach to public health care in Malaysia. AB - Both in its articulation of values and through incremental changes, the Malaysian government has signalled a change in attitude towards the welfare approach which had hitherto characterized public health care policy. This change envisions an end to reliance upon the state for the provision and financing of health services and the fostering of a system of family-based welfare. In the future citizens should finance their own health care through savings, insurance or as part of their terms of employment. While the state will still accept a degree of responsibility for those unable to pay for their health care, it wishes to share this burden with the corporate sector and non-government organizations as part of a national policy of the 'Caring Society'. In this article the retreat from a commitment to a welfare model of public health care is documented and some of the serious obstacles to such a policy are discussed. It is concluded that the government's aspirations for reforming the welfare model will need to be tempered by both practical and political considerations. Moreover, the socio-economic consequences of the Asian currency crisis of 1997 are likely to increase the need for government welfare action. PMID- 10387812 TI - Facility profile. Noises off: nursery pumps down the volume. PMID- 10387811 TI - Health sector reform and the interpretation of policy context. AB - Health sector reform is on the policy agenda of national governments and international agencies. This paper focuses on the policy process of health sector reform and, more specifically, the policy context. The importance of understanding policy context is emphasised and the elements of policy context are discussed. These are: demographic and epidemiological change; processes of social and economic change; economic and financial policy; politics and the political regime; ideology, public policy and the public sector. The paper then discusses the means for, and methods of, interpretation of the policy context. Particular emphasis is placed on linking policy context with an overall understanding of the policy process, the 'messiness' of policy-making, the interrelationship between the contextual factors, the contextual factors interpreted by policy actors, the use of analytical categories in the policy analysis. The paper concludes on the importance of strengthening policy analysis for effective health sector reform. PMID- 10387813 TI - Join the 'in' crowd. Want to keep your department in-house? Act like an outsourcer. PMID- 10387814 TI - Dare to compare. Do your housekeeping costs measure up? PMID- 10387815 TI - Reading the future. Two books will impact 21st century design. PMID- 10387816 TI - Drug prices edging upward again. PMID- 10387817 TI - Latex-free means costly; glove buy a balancing act. PMID- 10387818 TI - Give logistics its own place in the price equation. PMID- 10387819 TI - Mercy eyes better use of Premier's dual-source deals. PMID- 10387820 TI - Hospitals start to hoard supplies as Y2K looms. PMID- 10387821 TI - Clamp it! PMID- 10387822 TI - 1998 JEMS salary survey. Wages & benefits continue to decline. AB - Overall, EMS agencies are offering lower wages and fewer benefits today than they did five years ago, but overtime opportunities have increased for those willing to put in the extra time. And hard-working dispatchers can take heart: More systems are finally recognizing the key role dispatchers play in the delivery of high-quality EMS. PMID- 10387823 TI - Tactical medics. Front-line medicine evolves as a specialty. PMID- 10387825 TI - Whose license is it anyway? PMID- 10387824 TI - Letting go. Is it time to release your certification? PMID- 10387826 TI - Small claim, big malpractice headache. PMID- 10387827 TI - Keeping satellite-office doctors in the loop. PMID- 10387828 TI - Specialty carveouts: how big a threat to primary care? PMID- 10387829 TI - We need a new ICD-9: disease and death by tobacco. PMID- 10387830 TI - Could you be blamed for a patient's suicide? PMID- 10387831 TI - Resident matches. Is the primary care bandwagon running out of gas? PMID- 10387832 TI - Don't let this revolution leave you behind. PMID- 10387833 TI - "Doc, I saw this great new drug on TV...". PMID- 10387834 TI - Wrestling with the managed care octopus, Part 2. Stay up to date on patients' health benefits. PMID- 10387835 TI - Hospitals criticize HMO bailout plan. PMID- 10387836 TI - Upping the ante. Health plan betting on joint venture with Blues to change its luck in Atlantic City market. PMID- 10387837 TI - Feds: price of privacy may be too high. FBI, Justice Department say patient confidentiality bills would threaten fraud crackdown. PMID- 10387838 TI - Columbia hits snag in sale of Fla. hospitals. Buyer's rival asks state attorney general to review deal for possible antitrust violations. PMID- 10387839 TI - Going up. Prices rise after hospitals merge, no matter the ownership. PMID- 10387840 TI - Trial of Columbia executives to begin. PMID- 10387841 TI - We're Main Street, not Rodeo Drive. Rehab medicine is not a 'boutique' service; even bottom-line benefits were evident long ago. PMID- 10387842 TI - Launching the baby Columbias. Birth of spinoff chains LifePoint and Triad marks company's new focus on core operations. AB - This week marks a turning point for Columbia/HCA Healthcare Corp. The company that just a few years ago was gobbling up hospitals is now set to shed 56 facilities through two spinoffs-Triad Hospitals and LifePoint Hospitals. But observers say the spinoffs are good news for both Columbia and its offspring. PMID- 10387843 TI - Teaching hospitals bemoan lower margins. AAMC study shows Medicare spending limits will hit another healthcare provider group. PMID- 10387844 TI - Integration sacrificed for Y2K preparation. PMID- 10387845 TI - CHA guide gives members PR blueprint. PMID- 10387846 TI - Hospital companies bouncing back. Stock prices and investor confidence climb after chains release favorable earnings reports. PMID- 10387847 TI - Battle lines drawn. Medicare fight may hinge on whether private plans can win market share from government. PMID- 10387848 TI - Quality a concern with assisted living. PMID- 10387849 TI - Laying blame. GAO faults managed-care plans for Medicare problems; plans disagree, predict more pullouts. PMID- 10387850 TI - Medicare shifting costs to VA ... so VA seeking money from Medicare. PMID- 10387851 TI - Contract rehab firms closing up shop. PMID- 10387852 TI - Hospital lobbies vie for legislative relief. PMID- 10387854 TI - Quarterly mergers and acquisitions decline. PMID- 10387853 TI - Columbia business practices go on trial. Case begins against four execs accused of fraud for overcharging on Medicare cost reports. PMID- 10387855 TI - Premium jumps boost managed-care giants. PMID- 10387856 TI - Leading jointly. Two executives at Minnesota's Allina Health System show that co leadership can work. PMID- 10387857 TI - Fla. CEO, hospital in hot water. Exec fired over illegal campaign contributions; hospital could lose tax-exempt status. PMID- 10387858 TI - TB outbreak fallout. ER doc who contracted disease sues hospital for lack of warning, precautions. PMID- 10387859 TI - A dealmaker's biggest test. Democrat Breaux aims to shepherd 'premium support' plan through a skeptical Congress. Interview by Jonathan Gardner. PMID- 10387860 TI - Time for the what-if strategies. Still trying to eradicate remnants of the millennium bug, hospitals have some old-fashioned backup plans. AB - With time running out for healthcare providers to eradicate the millennium bug from their operations, they're now making sure contingency plans are in place so that regardless of the failures they encounter the week of Jan. 1, they'll still be able to remain open for business. PMID- 10387861 TI - Naming casualties. AHA ad campaign aims to show impact of Medicare spending restraints on local hospitals. PMID- 10387862 TI - A river runs through it. In the Richmond, Va., healthcare market, it's still north vs. south as for-profits line up against not-for-profits. PMID- 10387863 TI - Drug wars. Health plans blast Calif. regulators for requiring them to retain medications on their formularies. PMID- 10387864 TI - Tenn. system undergoes CEO exodus. Former hospital chief execs say they were casualties of NetCare's strategy to impress investors. PMID- 10387865 TI - MedPartners regains California network. PMID- 10387866 TI - Study: doc groups make Y2K progress. PMID- 10387868 TI - Georgetown settles PATH (Physicians at Teaching Hospitals) investigation. PMID- 10387867 TI - All quiet on the senior front. A powerful lobby is skeptical about providers' campaign to increase federal payments. PMID- 10387869 TI - Estimated losses from outpatient PPS rise. PMID- 10387870 TI - Medicare-risk retreats. Cigna to exit six HMO urban markets; Humana slows marketing. PMID- 10387871 TI - Slow growing for inpatient payments. PMID- 10387872 TI - Brazilian healthcare at a crossroads. Private sector flourishes as the government's program buckles under heavy demand, lack of funding. AB - In Brazilian healthcare today, almost anything goes. The government has a mandate to provide healthcare for all, but a dire lack of funding means coverage goes to only a few. Private managed-care plans have stepped in, turning the market wild and woolly. This story leads off a special international section. PMID- 10387873 TI - Brazil's info systems need a lift. Public health system reform and other changes are creating opportunities for new marketplace. PMID- 10387875 TI - Catholic leaders worldwide to meet. PMID- 10387874 TI - PPO deals in U.K. draw scrutiny. London hospital says it was "blacklisted" from Columbia insurance joint venture in Britain. PMID- 10387877 TI - Canadian icebreaker. Acute-care system succeeds with private bond issue. PMID- 10387876 TI - A changing long-term-care culture. To U.S. developers, Chile is a country prime for more eldercare facilities. PMID- 10387878 TI - What's your backup plan for turn of the new year? PMID- 10387879 TI - Recruiting for OR tougher as nursing shortage looms. PMID- 10387880 TI - GPOs offering reuse agreements. PMID- 10387881 TI - Culture, teamwork help better performers shine. PMID- 10387882 TI - Top performance across a system. PMID- 10387883 TI - Safety in laparoscopic electrosurgery. PMID- 10387884 TI - The use of 'surgical glue' spreads. PMID- 10387885 TI - New ORs built to adapt to change. PMID- 10387886 TI - Should OSHA issue latex bulletin? PMID- 10387887 TI - HCFA issues payment instructions for teleconsultations in rural HPSAs (health professional shortage areas). PMID- 10387888 TI - Collect what you're due under special managed care contract clauses. PMID- 10387889 TI - Several cancer procedures approved for Medicare coverage. PMID- 10387891 TI - What's up, doc.com? Health care moves closer to the digital age with the merger of WebMD and Healtheon. PMID- 10387890 TI - Duke's hazards. Did medical experiments put patients needlessly at risk? PMID- 10387892 TI - Do livestock breed drug-resistant bugs? Antibiotics on farms may threaten humans. PMID- 10387893 TI - The promise and peril of stem cell research. Scientists confront thorny ethical issues. PMID- 10387894 TI - Keeping breast cancer at bay. Women at risk are asked to weigh the pros and cons of a prevention pill. PMID- 10387896 TI - A national day only surgery benchmarking basket. AB - The efficient management of day surgery facilities benefits both patients and health administrators. Patients can benefit through minimisation of hospital stay while day surgery has the potential to increase elective surgery throughput and to reduce waiting times. This paper explores whether routinely collected morbidity data from Queensland public hospitals can be used to benchmark levels of day only surgery between hospitals. Thirteen procedures were identified that met criteria for inclusion in a day only surgery benchmarking basket. Queensland public hospitals and individual procedures were benchmarked against one another and analysed to determine whether hospitals performing the 13 procedures demonstrate the same rates of day only surgery. With the development of a clinically meaningful and administratively simple tool for comparing hospital day surgery rates using routinely collected morbidity data, the opportunity now exists for health services to compare the performance of clinical services both within and between hospitals. It is also suggested that the basket of procedures identified in this study could form the basis of a national day only surgery benchmarking process. PMID- 10387895 TI - Nursing as a career choice: perceptions of school students speaking Arabic, Serbo Croatian, Spanish, Turkish or Vietnamese at home. AB - Australia is a multicultural society and nowhere is this more evident than in Sydney where 25% of the population speaks a language other than English. In one of the largest area health services in New South Wales, the five most frequently spoken languages at home are Arabic, Serbo-Croatian, Spanish, Turkish or Vietnamese, with these language groups comprising 12% of Sydney's population. Yet nurses speaking one of these five languages comprise less than 1% of the nursing workforce. A cost-effective method of addressing the shortage of nurses speaking languages other than English is to recruit students who already speak another language into the profession. This study examined high school students' perceptions of nursing in order to determine appropriate methods of recruiting students speaking one of these languages. Implications for the design of recruitment campaigns are also discussed. PMID- 10387897 TI - Forecasting hospital expenditure in Victoria: lessons from Europe and Canada. AB - This paper specifies an econometric model to forecast State government expenditure on recognised public hospitals in Victoria. The OECD's recent cross country econometric work exploring factors affecting health spending was instructive. The model found that Victorian Gross State Product, population aged under 4 years, the mix of public and private patients in public hospitals, introduction of casemix funding and funding cuts, the proportion of public beds to total beds in Victoria and technology significantly impacted on expenditure. The model may have application internationally for forecasting health costs, particularly in short and medium-term budgetary cycles. PMID- 10387898 TI - Out with the old, in with the young: lifetime community rating. AB - Lifetime community rating has some potential benefits to private insurers, but they can only be realised if there is much greater control over private care providers than is currently the case. There is reason to fear that insurers' initial gains will disappear through increased provision of marginal care. Some members will gain through reduced premiums, and the main benefits will be derived by people who continue to maintain insurance. Most members will benefit hardly at all, and some (and particularly those who were unwilling or unable to take out insurance when they were young) will be significant losers. The public health care sector will remain under pressure at best, and it is more likely that the pressures will increase. The majority of Australians who do not have insurance will tend to lose. The obvious winners are the private care providers. The overall revenues of private health insurers will be relatively higher than if lifetime community rating were not introduced, and most of that revenue ultimately finds its way into the private care providers' pockets. Assuming they are able to increase the level of marginally useful care, there could be an increase in profitability to the extent that marginally useful care is actually less expensive to deliver. Finally, the government will derive another Pyrrhic victory. It will reduce its own outlays, but cause a decline in overall cost effectiveness of the health system. We have been here before, most recently in the period leading up to passage of the 30% rebate. There is good reason, therefore, to expect that lifetime community rating will be implemented. At least, the government will be able to claim it is defending Medicare from the more extreme privatisation ideas of Premier Kennett. This kind of argument will probably be sufficient. If so, the government will no doubt be stimulated to move to the next stage of dismantling of Medicare (which will presumably be something like means-testing of public hospital services). Many people believe that this is not an achievable goal in the near future. However, there was a popular view that the GST was not implementable after it lost the Coalition one election and led to Prime Minister Howard stating that he would 'never ever' raise the possibility again. The electorate is a sleeping giant, as is the public health care sector. It would be useful to know what could possibly serve as a wake-up call. Lifetime community rating is a small matter in the general trend towards killing off Medicare. But it is never too soon to send a message. PMID- 10387899 TI - National indicators for monitoring diabetes mellitus. PMID- 10387900 TI - Better theatre management through intelligent reporting: the TIME (theatre information, management and efficiency) system. AB - Hospital managers, clinicians or their colleges, and the government departments of health are interested in a variety of information for understanding the performance of the health care system and making informed decisions. Intelligent reporting aims to provide the most relevant and reliable information to major stakeholders to facilitate evidence-based practice. The key element in the reporting system is its ability to identify unsatisfactory practice. The TIME (theatre information, management and efficiency) system developed at the North Queensland Clinical School and James Cook University aims to provide intelligible reports for better theatre management. It reflects an effective amalgamation of surgical expertise and systems management principles. PMID- 10387901 TI - Episiotomy in NSW hospitals 1993-1996: towards understanding variations between public and private hospitals. AB - Episiotomy rates for women experiencing childbirth in New South Wales (NSW) hospitals are another indicator that private insurance may be a risk factor for obstetric intervention. A recent comparison of episiotomy rates in NSW public and private hospitals between 1993 and 1996 revealed that episiotomy rates were 12 to 15 percentage points higher in NSW private hospitals than in public hospitals studied. Rates also appear to be declining in NSW public hospitals, yet this trend is not evident in the NSW private hospitals studied. Although private hospital patients were almost twice as likely to experience forceps or vacuum delivery (often associated with episiotomy), this leaves a 6 to 8 percentage point difference unexplained. Given the potential health-related quality of life issues associated with perineal trauma during childbirth, further analysis of the clinical make-up of privately insured women may help determine the extent to which clinical explanations exist to support the differences in this childbirth intervention. PMID- 10387902 TI - Operating theatre information systems. PMID- 10387903 TI - Sorting out health financing: on personal choice and community good. PMID- 10387904 TI - When knowing the answer is only half the solution: introducing changes to a mental health rehabilitation service. AB - An Action Research methodology was used to revise the St George Mental Health Rehabilitation Service. A review committee found several limitations in the existing system, and a new model to address these problems was devised. During an 18-month period, the new model was implemented. Obstacles to change, such as staff resistance, were not systematically encountered due to the method of change used. The organisational changes that emerged concurrently improved the standard of service, raised staff morale, resulted in the routine use of standardised client outcome measures, enhanced the professional status of rehabilitation workers and led to empowerment of staff and consumers. PMID- 10387905 TI - Unintended outcomes of health care delivery and the need for a national risk management approach. AB - Unlike the situation in occupational health and safety, there is no nationally coordinated approach to risk management to prevent unintended outcomes of health care provision and improve health care quality. There is a related absence of linkages between quality assurance process, other programs aimed at patient injury prevention and professional indemnity insurance systems. This article discusses the Australian health policy direction and argues for a coordinated national health risk management approach developed through contract requirements which include duty of care and information provision, nationally approved standards and codes, professional liability requirements, and supporting health education, research and information technology development. PMID- 10387906 TI - Occupational stress in the operating theatre suite: should employers be concerned? AB - Research conducted amongst perioperative nurses during 1996 investigated both the causes of occupational stress and nurses' perceptions of the effects of modern medical technologies on several aspects of their work life during the preceding three years. It found that there was a strong perception amongst the 433 nurses in the study that medical technologies had contributed to their increased workloads and higher levels of stress. This article presents the key findings on occupational stress and discusses some of their implications for health service managers who have responsibility for the occupational health and safety of nurses working in the operating theatre environment. PMID- 10387907 TI - Personal choices on private health insurance. PMID- 10387908 TI - Health sector liability under the Trade Practices Act. AB - Following the implementation of the national competition policy and the consequent exposure of unincorporated businesses to trade practices regulation, the health sector has faced increasing exposure to fair trading and competition issues. This article examines the rights and the obligations of health sector participants under the consumer protection and the restrictive trade practices provisions of the Trade Practices Act 1974 (Cwlth). The article outlines the relevant provisions and identifies examples of conduct that has breached the Act or that has the potential to breach the Act. The author notes that the Act has been applied to the health sector in the same way as it has been applied to all other sectors of the economy. PMID- 10387909 TI - The effects of income inequality on health. AB - Much of the discussion about individual and group differences in illness and life expectancy has focused on the effects of individual characteristics, both status and behavioural. This is also characteristic of much of the literature, which attempts to explain why men have higher rates of disease and lower life expectancy than women. After a period in which 'social policy was no longer such an important part of preventive health policy', there is now renewed interest in the influence of the socioeconomic environment on health. Indeed, recently compiled evidence indicates that increasing income inequality is likely to have adverse effects on the community's health. These findings highlight the potential dangers of policy changes which accelerate social and economic divisions. PMID- 10387910 TI - Leadership for a healthy 21st century. AB - Every economic institution finds itself caught on the horns of a dilemma: Competing sets of values strike a conflict between social good and economic wealth, regardless of whether organizations articulate it. The struggle in U.S. health care, however, is both more acute and poignant. On the one hand, ethical and cultural values require societal commitments to the well-being of the individual. Who among us would want to refuse help to someone sick or injured? On the other hand, market forces require an economic accounting of health care. Social trends emphasize a mission to provide care for all, while managed care promotes the industry's fidelity to a balanced ledger. U.S. health care is thus defined by paradox. The nation spends more than $1.3 trillion annually on health care--a national line-item larger than the economies of all other nations except two (Germany and Japan). A new study by the Health Care Financing Administration warns that health care spending may nearly double to $2.1 trillion by 2007. Yet the industry is perceived to be too "resource-constrained" to assure health care services for all citizens. This poses a key question: Are there too few resources, or are we simply not allocating them in the best ways possible? Health care's "double bottom line"--social and economic accountability--typifies the social and economic milieu of health care as the final pages turn on the 20th Century. And, it is this duality that forms the underlying theme for the landmark study--Leadership for a Healthy 21st Century--conducted over the course of the past year by Arthur Andersen and The Healthcare Forum Foundation, with primary research support from DYG, Inc. and Baruch Lev, professor at the Stern School of Business, New York University. The study was designed to investigate a new economic model emerging in the information economy and its impact on health care; the evolving values of consumers in relation to business, health and health care; and the values of corporate leaders in health care and other industries. The overall objective: development of a 21st Century leadership model in health care capable of resolving the conflicts inherent in serving two masters: (1) compassion and (2) wealth and value creation. PMID- 10387911 TI - Sixteen ways to blur your organization: a conversation with Chris Meyer. Interview by Joe Flower. PMID- 10387912 TI - Seven practices of successful organizations. Part 1: Employment security, selective hiring, self-managed teams, high compensation. PMID- 10387913 TI - A six-step model for today's healthcare competitors. PMID- 10387914 TI - Will you be ready for HMOs 'R'Us? PMID- 10387916 TI - Painting the future picture: changing an organization by moving beyond its comfort zone. PMID- 10387915 TI - Second-wave relationships: the quest for revenue. PMID- 10387917 TI - The crash of ValuJet 592: implications for healthcare. PMID- 10387918 TI - Therapeutic breakthroughs in the new millennium: what to look for in the next two decades. Part 1: New answers for cancer, drug resistance, food poisoning. PMID- 10387919 TI - Truth or consequences. PMID- 10387920 TI - The NHS Estates Healthcare Estate Management Awards 1998. PMID- 10387921 TI - An end to final salary pension plans in the public sector? PMID- 10387922 TI - Flat demand for private medical insurance in 1997. PMID- 10387923 TI - Hospital occupancy costs. PMID- 10387924 TI - Filtration for hospital and medical applications. PMID- 10387925 TI - Microbiologically induced corrosion. PMID- 10387926 TI - The existence of the 4S polycyclic aromatic hydrocarbon-protein binding in 14-day old chick embryo liver. AB - Cytochrome P-450IA1, the isozyme most closely associated with aryl hydrocarbon hydroxylase (AHH), is regulated by two high-affinity binding proteins, the 4S polycyclic aromatic hydrocarbon (PAH)-binding protein which primarily binds PAHs and the 8S Ah (dioxin) receptor which binds 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and like congeners. The present study was conducted to determine whether the 4S protein existed in 14-day-old chick embryo liver when AHH activity is maximal to determine if they are linked as is the 8S Ah receptor and to confirm the existence of the dioxin receptor by investigating their ligand binding characteristics in the presence and absence of sodium molybdate, an agent that stabilizes steroid hormone receptors and partially stabilizes the dioxin receptor. Competitive ligand binding studies were performed with liver cytosol from livers of male 14-day-old chick embryos using [3H]-benzo[a]pyrene (B[a]P) or [3H]-TCDD in the presence and absence of a 200-fold excess of B[a]P, benzo[e]pyrene (B[e]P), 3-methylcholanthrene (3-MC), and tetrachlorodibenzofuran (TCDBF). Specific PAH-binding activity was assayed using sucrose gradient analysis. In the absence of molybdate, the 4S PAH-binding protein had high affinity for B[a]P, B[e]P, 3-MC, but very low affinity for TCDBF; the Ah receptor exhibited high affinity for TCDBF. In the presence of sodium molybdate, the Ah receptor was stabilized while the 4S PAH-binding protein was relatively unaffected. These results affirm the existence of two distinct PAH-binding proteins in 14-day-old chick embryo liver cytosol and suggest a linkage of the 4S protein to AHH. PMID- 10387927 TI - Enzymatic methylation of arsenic compounds. VII. Monomethylarsonous acid (MMAIII) is the substrate for MMA methyltransferase of rabbit liver and human hepatocytes. AB - Inorganic arsenite is methylated by some, but not all, animal species to dimethylarsinic acid (DMA). The monomethyl compound containing arsenic in an oxidation state of +3 has been proposed as an intermediate. Using highly purified arsenic methyltransferase from rabbit liver and the partially purified enzyme from Chang human liver hepatocytes, the activity of methylarsonic acid (MMAV) and methylarsonous acid (MMAIII) as a substrate has been characterized by Michaelis Menten kinetics. The rabbit liver enzyme has a greater affinity for MMAIII (Km = 0.92 x 10(-5) M) than MMAV (Km = 7.0 x 10(-5) M) since the smaller the Km the greater the affinity. In addition, a dithiol, reduced lipoic acid or dithiothreitol, appears to be more active than GSH in satisfying the thiol requirement of the enzyme. Although investigators have been unable to detect the arsenic methyltransferase in surgically removed human liver, its presence in Chang human hepatocytes now has been established. The Km for MMAIII, 3.04 x 10( 6), using MMAIII methyltransferase from Chang human hepatocytes was not greatly different from that of the rabbit liver enzyme. PMID- 10387928 TI - Gestational exposure to chlorpyrifos: comparative distribution of trichloropyridinol in the fetus and dam. AB - Chlorpyrifos (O,O'-diethyl O-[3,5,6-trichloro-2-pyridyl] phosphorothionate) is a commonly used anticholinesterase insecticide, and therefore the potential for human exposure is high. The present time course and dose response studies were conducted to delineate the toxicokinetics of chlorpyrifos and its metabolites in the pregnant rat and fetus. Time-pregnant, Long-Evans rats were treated orally with chlorpyrifos during late gestation (Gestational Days 14-18). Following euthanasia the level of chlorpyrifos and its metabolites, chlorpyrifos-oxon and 3,5,6-trichloro-2-pyridinol (TCP), were measured in both fetal and maternal brain and liver (limits of quantitation: 59.2, 28.8, and 14.0 ng/g tissue, respectively). In addition, cholinesterase inhibition was also measured in the same tissues for comparison. TCP was the only component detected. The highest level of TCP and the lowest level of cholinesterase activity showed the same time of peak effect: 5 h after the last dose. The concentration of TCP in the maternal liver was approximately fivefold higher than the TCP concentration in fetal liver, but, paradoxically, the concentration of TCP in the fetal brain was two- to fourfold higher than the TCP concentration in the maternal brain. The half life of the TCP was identical in all tissues examined (12-15 h). These toxicokinetic results suggest that the fetal nervous system may be exposed to a higher concentration of chlorpyrifos than the maternal nervous system when the dam is orally exposed to chlorpyrifos during late gestation. PMID- 10387929 TI - Uterotrophic effects of tamoxifen, toremifene, and raloxifene do not predict endometrial cell proliferation in the ovariectomized CD1 mouse. AB - The uterotrophic responses of ovariectomized CD1 mice to tamoxifen, toremifene, and raloxifene have been compared to 17beta-estradiol after a treatment period of 72 h. Uterine and vaginal weight, luminal epithelial thickening, and 5 bromodeoxyuridine (BrdU) labeling index in the endometrial stroma were examined. All three pharmaceuticals, as well as 17beta-estradiol, produced increases in the classic estrogen-dependent variables of uterine and vaginal weights after the 3 day treatment period. Tamoxifen, toremifene, raloxifene, and estradiol all increased luminal epithelial thickness, and increased the BrdU labeling index in the endometrial stroma of the uterus. Although the dose response for the uterotrophic effect and the vaginal weight increases for toremifene differed from tamoxifen and raloxifene, in that there was no dose at which these effects were maximal, the stimulation of BrdU labeling index in the endometrial stroma was dose dependent and very similar for all three, at the clinically relevant doses. Treatment-related hypertrophic effects were estimated by examination of the nuclear profile density in the endometrial stroma. Estradiol and tamoxifen caused a greater hypertrophic effect than toremifene and raloxifene, indicating that factors other than an increase in cell number contribute to the overall uterotrophic effect. This demonstrates that the use of uterine weight to estimate the relative estrogenicity of drugs could give a misleading impression of the response of the uterus to estrogen agonists. Variables, such as increased DNA replication, which may be more important to a subsequent potential carcinogenic process in the uterus, for a particular drug, requires separate evaluation. PMID- 10387930 TI - A chimeric aryl hydrocarbon receptor knockout mouse model indicates that aryl hydrocarbon receptor activation in hematopoietic cells contributes to the hepatic lesions induced by 2,3,7, 8-tetrachlorodibenzo-p-dioxin. AB - Pathologic changes associated with 2,3,7, 8-tetrachlorodibenzo-p-dioxin (TCDD) exposure have been reported in the livers of a wide range of species. While these changes have been extensively described, the mechanisms of toxic interaction(s) that produce these lesions remain unclear. Using an aryl hydrocarbon receptor (Ahr) knockout male mouse chimeric model, we investigated whether the presence of this receptor in hematopoietic and/or parenchymal cells affects TCDD-induced hepatotoxicity. Bone marrow chimeras were produced by hematopoietic reconstitution of irradiated mice. Specifically, chimeras were generated with aryl hydrocarbon receptor (AHR) positive hematopoietic and parenchymal cells (Ahr+/+ animal bone marrow cells into irradiated Ahr+/+ animals), AHR positive hematopoietic and negative parenchymal cells (Ahr+/+ into Ahr-/-), AHR negative hematopoietic and positive parenchymal cells (Ahr-/- into Ahr+/+), and AHR negative hematopoietic and parenchymal cells (Ahr-/- into Ahr-/-). Male wild-type (Ahr+/+) and knockout (Ahr-/-) animals were used as nonchimeric controls. Following TCDD treatment (30 microg/kg body wt), liver sections from mice in each control and chimeric group were histologically evaluated for necrotic and inflammatory changes. TCDD treatment produced moderate inflammation in Ahr+/+ controls and Ahr+/+ into Ahr+/+ chimeras. This response was mild in TCDD-treated Ahr-/-, Ahr-/- into Ahr-/-, Ahr+/+ into Ahr-/-, and Ahr-/- into Ahr+/+ animals and was not different from the corresponding vehicle-treated groups. Moderate necrosis was observed in all TCDD-treated controls or chimeras with AHR-positive parenchyma. No or mild necrosis was observed in TCDD- and vehicle-treated animals containing AHR-negative parenchyma. These data indicate that the presence of AHR in hepatic parenchyma alone is sufficient for TCDD induction of hepatic necrosis, and its presence in hematopoietic cells is necessary for the inflammatory response to TCDD-induced hepatic lesions. PMID- 10387931 TI - Effect of lead exposure and accumulation on copper homeostasis in cultured C6 rat glioma cells. AB - C6 rat glioma cells resemble rat astroglia in culture in that both cell types accumulate lead (Pb) intracellularly from the medium. As such, C6 cells are a model for Pb accumulation by the brain. In this study, an increase in intracellular Pb accumulation induced by p-chloromercuribenzoate (PCMB) after exposure to 10 microM Pb acetate suggests a role for sulfhydryl groups in Pb retention. Stimulation of Pb accumulation by nifedipine suggests the entry of Pb into these cells by a novel path. Most of the intracellular Pb from exposure for 7 days to 1 microM Pb was associated with high-molecular weight components in cytosol. Pb exposure increased the abundance of three proteins with the following characteristics on two-dimensional gels: 81 kDa with pI of 5.6, 81 kDa with pI of 4. 9, and 71 kDa with pI of 5.6. The levels of five other proteins, ranging in size from 37-41 kDa with pIs of 6.0-6.8 declined. Exposed C6 cells accumulated copper (Cu) intracellularly, and Cu accumulation after Pb exposure was shown by kinetic analysis with 67Cu to result from an increased uptake and a decreased efflux for Cu. Pb-exposed cells also showed increased Cu binding to membranes, which is consistent with the increase of Cu uptake. These data indicate that intracellular Pb interacts with high molecular weight proteins in C6 cells, and exposure also alters membrane transport properties for copper. PMID- 10387932 TI - Structure-hepatic disposition relationships for phenolic compounds. AB - Phenolic compounds are widely used in therapeutic, environmental, and industrial applications. The present work seeks to define the hepatic disposition of 11 phenolic compounds with varying lipophilicities and molecular weights. The hepatic disposition kinetics were studied in a once-through in situ rat liver perfusion preparation in order to avoid extra-hepatic metabolism and recirculation effects. The phenols were administered using the impulse-response technique and the time course of hepatic venous effluent concentration was examined by moments and a two-compartment dispersion model. While the extraction of the phenolic compounds was relatively independent of lipophilicity, the estimated permeability-surface area (PS) product for influx of solutes into the hepatocytes could be related to the compounds' octanol-buffer partition coefficients (log Papp). This log PS-logPapp relationship was consistent with that reported earlier for another series of solutes with a wide range of lipophilicity. The metabolites produced from each of the phenolic compounds used in this study had mean transit times similar to those of their corresponding parent phenols, suggesting that the metabolites were not trapped in the liver as a consequence of their higher polarity. It is concluded that the strong solute lipophilicity-toxicity and lipophilicity-skin penetration relationships often seen for aqueous solutions of phenols are not evident for the hepatic extraction of these compounds. Such a conclusion is consistent with the hepatic extraction of phenolic compounds being mainly determined by a blood flow limitation in delivery of the phenol to the liver, rather than the intrinsic liver metabolic enzyme activities at the doses injected. PMID- 10387933 TI - Quantitative measurement of acute corneal injury in rabbits with surfactants of different type and irritancy. AB - We have hypothesized that differences in ocular irritancy are related to differences in extent of initial injury and that, regardless of the processes leading to tissue damage, extent of injury is the primary factor that determines the final outcome of ocular irritation. In previous in vivo confocal microscopic (CM) studies we identified quantifiable differences in the extent of corneal injury occurring with four surfactants (three anionic, one cationic) known to cause different levels of ocular irritation and demonstrated that extent of initial corneal injury was related to the magnitude of cell death. The purpose of this study was to assess the applicability of this hypothesis to a broad sampling of surfactants. Specifically, initial corneal changes induced by seven different surfactants (one anionic, three cationic, three nonionic) were measured by in vivo CM and cell death was measured by an ex vivo live/dead assay. The right eye of each rabbit was treated by placing 10 microl of a surfactant directly on the cornea. Eyes were examined macroscopically and scored for irritation at 3 h and 1 day. At 3 h and 1 day, in vivo CM was used to examine the corneas and quantitate epithelial cell size, epithelial thickness, corneal thickness, and depth of stromal injury. At 3 h and/or at 1 day, corneas were removed and excised regions were placed in culture media containing 2 microM calcein AM and 4 microM ethidium homodimer. Using laser scanning CM, the number of dead epithelial and/or stromal cells in a 300 x 300 x 170-microm3 (xyz) volume of the cornea was determined. In vivo CM and live/dead assay findings revealed three surfactants to affect only the epithelium, three surfactants to affect the epithelium and superficial stroma, and one surfactant to affect the epithelium and deep stroma. Extent of initial corneal injury reflected level of ocular irritation, and magnitude of cell death was related to the extent of initial corneal injury. These findings are consistent with those for known slight, mild, and moderate to severe irritants, respectively. They suggest that our hypothesis is broadly applicable to surfactants. Additionally, we believe these surfactants should be included as part of a new "gold standard" for use in developing and validating in vitro tests to replace the use of animals in ocular irritancy testing. PMID- 10387934 TI - Peroxisome-proliferator regulates key enzymes of the tryptophan-NAD+ pathway. AB - Structually diverse peroxisome-proliferators (PPs) were investigated regarding their effects on NAD+ level and two key enzyme activities in the tryptophan (Trp) NAD+ pathway in the liver of rats (Sprague-Dawley male) fed PP-containing diets freely for 2 weeks. All PPs, except for thyroxine, significantly increased hepatic NAD+ level in concert with hepatic hypertrophy. Activity of quinolinate phosphoribosyltransferase (QAPRTase), one of the key enzymes in the Trp-NAD+ pathway, was increased by the PPs which caused significant increase in the hepatic NAD+. On the other hand, alpha-amino-beta-carboxymuconate-epsilon semialdehyde decarboxylase (ACMSDase), another key enzyme in the Trp-NAD+ pathway, was drastically inhibited by all PPs except for linolenic acid, which was only slightly inhibitory. Most PPs investigated activated peroxisomal marker enzymes such as palmitoyl-CoA oxidase, catalase, and PPAR-alpha(peroxisome proliferator activated receptor-alpha)-dependent enzymes, such as malic enzyme and l-3-glycerophosphate dehydrogenase. NAD+ was also increased in the rat hepatocytes cultured in the medium supplemented with PPs. These data suggested that regulation of the key enzymes in the Trp-NAD+ pathway was associated with PPAR-alpha directly or indirectly, and as a consequence the hepatic NAD+ was increased by PPs. PMID- 10387935 TI - Prevalence and clinical outcome of mitral-valve prolapse. AB - BACKGROUND: Mitral-valve prolapse has been described as a common disease with frequent complications. To determine the prevalence of mitral-valve prolapse in the general population, as diagnosed with the use of current two-dimensional echocardiographic criteria, we examined the echocardiograms of 1845 women and 1646 men (mean [+/-SD] age, 54.7+/-10.0 years) who participated in the fifth examination of the offspring cohort of the Framingham Heart Study. METHODS: Classic mitral-valve prolapse was defined as superior displacement of the mitral leaflets of more than 2 mm during systole and as a maximal leaflet thickness of at least 5 mm during diastasis, and nonclassic prolapse was defined as displacement of more than 2 mm, with a maximal thickness of less than 5 mm. RESULTS: A total of 84 subjects (2.4 percent) had mitral-valve prolapse: 47 (1.3 percent) had classic prolapse, and 37 (1.1 percent) had nonclassic prolapse. Their age and sex distributions were similar to those of the subjects without prolapse. None of the subjects with prolapse had a history of heart failure, one (1.2 percent) had atrial fibrillation, one (1.2 percent) had cerebrovascular disease, and three (3.6 percent) had syncope, as compared with unadjusted prevalences of these findings in the subjects without prolapse of 0.7, 1.7, 1.5, and 3.0 percent, respectively. The frequencies of chest pain, dyspnea, and electrocardiographic abnormalities were similar among subjects with prolapse and those without prolapse. The subjects with prolapse were leaner (P<0.001) and had a greater degree of mitral regurgitation than those without prolapse, but on average the regurgitation was classified as trace or mild. CONCLUSIONS: In a community based sample of the population, the prevalence of mitral-valve prolapse was lower than previously reported. The prevalence of adverse sequelae commonly associated with mitral-valve prolapse in studies of patients referred for that diagnosis was also low. PMID- 10387936 TI - Lack of evidence of an association between mitral-valve prolapse and stroke in young patients. AB - BACKGROUND: Previous studies have reported a high prevalence of mitral-valve prolapse among patients with embolic stroke (28 to 40 percent), especially among young patients (those < or =45 years old); this finding has practical implications for prophylaxis. However, diagnostic criteria for prolapse have changed and are now based on three-dimensional analysis of the shape of the valve; use of the current criteria reduces markedly the frequency of such a diagnosis and increases its specificity. Previously described complications must therefore be reconsidered. METHODS: In a case-control study, we reviewed data on 213 consecutive patients 45 years old or younger with documented ischemic stroke or transient ischemic attack between 1985 and 1995; they underwent complete neurologic and echocardiographic evaluations. The prevalence of prolapse in these patients was compared with that in 263 control subjects without known heart disease, who were referred to our institution for assessment of ventricular function before receiving chemotherapy. RESULTS: Mitral-valve prolapse was present in 4 of the 213 young patients with stroke (1.9 percent), as compared with 7 of the 263 controls (2.7 percent); prolapse was present in 2 of 71 patients (2.8 percent) with otherwise unexplained stroke. The crude odds ratio for mitral-valve prolapse among the patients who had strokes, as compared with those who did not have strokes, was 0.70 (95 percent confidence interval, 0.15 to 2.80; P=0.80); after adjustment for age and sex, the odds ratio was 0.59 (95 percent confidence interval, 0.12 to 2.50; P=0.62). CONCLUSIONS: Mitral-valve prolapse is considerably less common than previously reported among young patients with stroke or transient ischemic attack, including unexplained stroke, and no more common than among controls. Using more specific and currently accepted echocardiographic criteria, therefore, we could not demonstrate an association between the presence of mitral-valve prolapse and acute ischemic neurologic events in young people. PMID- 10387938 TI - Occult hepatitis B virus infection in patients with chronic hepatitis C liver disease. AB - BACKGROUND: Hepatitis B virus (HBV) infections in patients who lack detectable hepatitis B surface antigen (HBsAg) are called occult infections. Although such infections have been identified in patients with chronic hepatitis C liver disease, their prevalence and clinical significance are not known. METHODS: With the polymerase chain reaction, we searched for HBV DNA in liver and serum samples from 200 HBsAg-negative patients with hepatitis C virus (HCV)-related liver disease (147 with chronic hepatitis, 48 with cirrhosis, and 5 with minimal histologic changes). One hundred of the patients had detectable antibodies to the HBV core antigen (anti-HBc); 100 were negative for all HBV markers. Eighty-three were treated with interferon alfa. We also studied 50 patients with liver disease who were negative both for HBsAg and for HCV markers. In six patients found to have occult HBV infection, we evaluated possible genomic rearrangements through cloning or direct sequencing procedures. RESULTS: Sixty-six of the 200 patients with chronic hepatitis C liver disease (33 percent) had HBV sequences, as did 7 of the 50 patients with liver disease unrelated to hepatitis C (14 percent, P=0.01). Among the 66 patients, 46 were anti-HBc-positive and 20 were negative for all HBV markers (P<0.001). Twenty-two of these 66 patients (33 percent) had cirrhosis, as compared with 26 of the 134 patients with hepatitis C infection but no HBV sequences (19 percent, P=0.04). HBV sequences were detected in 26 of the 55 patients in whom interferon therapy was ineffective and 7 of the 28 patients in whom interferon therapy was effective (P=0.06). None of the sequenced HBV genomes had changes known to interfere with viral activity and gene expression. CONCLUSIONS: Occult hepatitis B infection occurs frequently in patients with chronic hepatitis C liver disease and may have clinical significance. PMID- 10387937 TI - Long-term survival and late deaths after allogeneic bone marrow transplantation. Late Effects Working Committee of the International Bone Marrow Transplant Registry. AB - BACKGROUND AND METHODS: It is uncertain whether mortality rates among patients who have undergone bone marrow transplantation return to the level of the mortality rates of the general population. We analyzed the characteristics of 6691 patients listed in the International Bone Marrow Transplant Registry. All the patients were free of their original disease two years after allogeneic bone marrow transplantation. Mortality rates in this cohort were compared with those of an age-, sex-, and nationality-matched general population. Cox proportional hazards regression was used to identify risk factors for death more than two years after transplantation (late death). RESULTS: Among patients who were free of disease two years after transplantation, the probability of living for five more years was 89 percent (95 percent confidence interval, 88 to 90 percent). Among patients who underwent transplantation for aplastic anemia, the risk of death by the sixth year after transplantation did not differ significantly from that of a normal population. Mortality remained significantly higher than normal throughout the study among patients who underwent transplantation for acute lymphoblastic leukemia or chronic myelogenous leukemia and through the ninth year among those who underwent transplantation for acute myelogenous leukemia. Recurrent leukemia was the chief cause of death among patients who received a transplant for leukemia, whereas chronic graft-versus-host disease was the chief cause among those who received a transplant for aplastic anemia. Advanced, long standing disease before transplantation and active chronic graft-versus-host disease were important risk factors for late death. CONCLUSIONS: In patients who receive an allogeneic bone marrow transplant as treatment for acute myelogenous or lymphoblastic leukemia, chronic myelogenous leukemia, or aplastic anemia and who are free of their original disease two years later, the disease is probably cured. However, for many years after transplantation, the mortality among these patients is higher than that in a normal population. PMID- 10387940 TI - Shattuck lecture--medical and societal consequences of the Human Genome Project. PMID- 10387939 TI - Images in clinical medicine. A grade 6 systolic murmur. PMID- 10387941 TI - Occult gastrointestinal bleeding. PMID- 10387942 TI - Primary Care Review Articles -- Practical Information for Generalist Physicians. PMID- 10387943 TI - Mechanisms of a phosphorothioate oligonucleotide delivery by skin electroporation. AB - Skin electroporation has great potential for topical delivery of oligonucleotides. Controled therapeutic levels of an intact phosphorothioate oligonucleotide (PS) can be reached in the viable tissue of the skin. The aim of this work was to investigate the transport mechanisms of a PS in hairless rat skin by electroporation, and hence to allow optimization of oligonucleotides (ONs) topical delivery. The pulsing condition used was five exponentially decaying pulses of 100 V and 500 ms pulse time. The main mechanism of PS transport in the skin viable tissues during pulsing was electrophoresis. The electroosmosis contribution was negligible. Electrophoresis created within minutes a reservoir of PS in the skin viable tissues, which persisted within a therapeutic range of hours. A strong PS/stratum corneum interaction occurred. PMID- 10387944 TI - Inclusion complex of piroxicam with beta-cyclodextrin and incorporation in hexadecyltrimethylammonium bromide based microemulsion. AB - The interaction of piroxicam with beta-cyclodextrin (beta-CD), hexadecyltrimethylammonium bromide-based microemulsion (ME), and ME in the presence of beta-CD aimed at the optimization of topical drug delivery was studied. UV-VIS absorption spectra at pH 5.5 were obtained with and without beta CD and ME. The stability constant (K) values for the piroxicam/beta-CD complex in the pH range 4.5-6.0 varied from 87 to 29 M-1. The cationic microemulsion was characterized by pseudo-ternary phase diagram. The association constant (Ks) of piroxicam/ME was determined using the framework of the pseudophase model. The value of Ks obtained for piroxicam at pH 5.5 was 132 M-1. At the same pH, the value of Ks for the incorporation of piroxicam/beta-CD complex in the ME was 150 M-1. PMID- 10387945 TI - Incorporating batch effects in the estimation of drug stability parameters using an Arrhenius model. AB - The nonlinear estimation of drug stability parameters (energy of activation Ea and shelf-life tY) by conventional approaches employs equations relating drug content determination C at time t and temperature T. The identification procedures lead to the determination of only one initial drug content C0 for several different experiments. However, it is well known that because of experimental concentration variation or of intentional modification of the experimental schedule, there are as many initial drug contents as experiments. For these reasons, a method which takes into account batch effects is proposed to determine stability parameters and also all initial drug contents C0j where j is the index of experiment in one step. This method is more accurate from a statistical viewpoint and is suitable for data treatment in pharmaceutical industries where the initial drug content of each batch entering the stability program can be checked a posteriori. The application of this method is shown on real kinetic data from the hydrolysis of acetylsalicylic acid (ASA). PMID- 10387946 TI - New cytokine dressings. I. Kinetics of the in vitro rhG-CSF, rhGM-CSF, and rhEGF release from the dressings. AB - Wound repair-stimulatory activities of various cytokines and growth factors depend on successful delivery of these factors to the injured sites. Here were present the design and preparation of the new collagen- and polyurethane-based dressings containing the recombinant human cytokines-rhG-CSF, rhGM-CSF or rhEGF. To test the efficacy of the retrieval of the incorporated cytokines, their controlled release from the dressings was carried out over three consecutive days using polyurethane sponge as a collector of the extracts. The maximum quantities of the released rhG-CSF, rhGM-CSF and rhEGF reached approximately 25, 50, and 10%, respectively, of the total cytokine contents of the dressings, as assessed by the specific ELISA tests. These data indicate that collagen- and polyurethane dressings containing rhGM-CSF and rhG/CSF may serve as effective tools for the topical delivery of cytokines to wounded tissues. PMID- 10387947 TI - New cytokine dressings. II. Stimulation of oxidative burst in leucocytes in vitro and reduction of viable bacteria within an infected wound. AB - Recently, we have developed the new dressings containing rhG-CSF or rhGM-CSF. In the present study we investigated either in vitro or in vivo biological activity of the dressings. Human whole blood samples were incubated with extracts from the collagen- or polyurethane-based dressings containing rhG-CSF or/and rhGM-CSF and phagocytic and oxidative metabolic activities were quantitated using Phagotest or Bursttest kits. The results indicate that both the number of phagocyting cells and the intensity of phagocytosis per cell, as well as the level of the oxidative burst in particular, were stimulated by one or both of the cytokines extracted. Next, the experimental skin wounds in mice were infected with 107 CFU of Pseudomonas aeruginosa strain ATCC 27853 and treated locally with the rhG-CSF containing dressing. The analysis of the biopsies taken from the wounds indicated that in mice treated with the cytokine-containing dressing on the third day the log of CFU per biopsy was 5.0 vs. 6.2 in the control (P<0.001), and on the 8th day was lower than 4 vs. 5.4 in control (P<0.0001). Our findings clearly suggest that the newly designed dressings containing the incorporated CSFs can be used as effective topical cytokine-delivery system in the treatment of bacterial infections in wounds. PMID- 10387948 TI - Absorption of insulin from pluronic F-127 gels following subcutaneous administration in rats. AB - The main objective of this work was to evaluate the use of Pluronic (PF127) gels, polylactic-co-glycolic acid (PLGA) nanoparticles and their combination for parenteral delivery of peptides and proteins having short half-lives using insulin as a model drug. The in vitro insulin release profiles of various PF127 formulations were evaluated at 37 degrees C using a membraneless in vitro model. In vivo evaluation of the serum glucose and insulin levels was performed following subcutaneous administration of various insulin formulations in normal rats. The in vitro results demonstrated that the higher the concentration of PF127 in the gel, the slower the release of insulin from the matrices, independent of the vehicle used. The acute hypoglycemic peak resulting from administration of an insulin solution between 0.5 and 2.0 h after administration (peak at 1 h) is replaced after administration of insulin-PLGA nanoparticles by an almost constant hypoglycemic effect with a slower recovery of the serum glucose levels at about 2 h after administration. By loading insulin into PF127 gels, a slower and more prolonged hypoglycemic effect of insulin was obtained in inverse proportion to the polymer concentration. PF127 gel formulations containing insulin-PLGA nanoparticles had the most long-lasting hypoglycemic effects of all formulations. From the current in vitro and in vivo study, we concluded that PF127 gel formulations containing either drug or drug nanoparticles could be useful for the preparation of a controlled delivery system for peptides and proteins having short half-lives. PMID- 10387949 TI - The use of isothermal heat conduction microcalorimetry to evaluate drug stability in tablets. AB - Isothermal heat conduction microcalorimetry was used to evaluate chemical stability of a solid drug in tablets. A variety of mixtures were compressed to flat faced tablets of 300 mg weight and 10 mm diameter. The content of drug amounted to 10%. Besides drug containing tablets, also placebo tablets as well as the non compressed mixtures were examined by microcalorimetry at 80 degrees C. The excipient Emcompress exhibited a substantially high exothermic heat flow that was due to a change in crystallinity. For Emcompress containing tablets this interfering signal resulted in such a way that the calorimetric data did not reflect the drug decomposition with sufficient accuracy. In the case of the other preparations the heat flow of the excipients were low, and the calorimetric data did reflect the drug decomposition. The stability increased with increasing content of CaHPO4, respectively, with decreasing content of water. PMID- 10387950 TI - An investigation into interactions between polyacrylic polymers and a non-ionic surfactant: an emulsion preformulation study. AB - The aim of this study was to investigate possible interactions between a polymeric emulsifier and a non-ionic surfactant, with a view of achieving better understanding of emulsion stabilisation mechanisms. The polymeric emulsifier used was acrylates/C10-30 alkyl acrylate crosspolymer (Pemulen TR-2(R)), while Polyoxyethylene 20 sorbitan mono-oleate (Polysorbate 80) has been chosen as a model surfactant. Both materials were used within the concentration range relevant for their practical application. A 0.2%w/w aqueous dispersion of polymeric emulsifier, containing various amounts of surfactant (from 0.01 to 1.0% w/w) was used throughout the study. Interfacial aspects of the proposed polymer/surfactant interactions were analysed by means of surface tension measurements. Changes in the network structure of the test dispersions were quantified by continuous shear rheometry, supported by the texture analysis. To analyse the influence of hydrophobic alkyl groups present on the Pemulen TR-2(R) chains, an unmodified, hydrophilic polyacrylic acid polymer, Carbopol 934P(R), was assessed under the same conditions. The results obtained by both surface tension and rheological studies have revealed large differences in behaviour of the two polymers in the presence of the model surfactant. Pemulen TR-2(R) was shown to desorb the surfactant from the surface, within the whole concentration range studied. Furthermore, an increase in viscosity and texture profile parameters with increasing Polysorbate 80 concentration up to 0.3% w/w was evident in the case of Pemulen TR-2(R) dispersions. This was followed by a decrease in the gel network strength at higher surfactant concentrations. On the other hand, Carbopol 934P(R) has shown no signs of surfactant desorption and only small changes in the network structure with the increasing concentration of surfactant. It is shown in this study that an interaction between a polymeric emulsifier Pemulen TR-2(R) and a non-ionic surfactant Polysorbate 80 does occur in their aqueous dispersion, and that it is: (a) hydrophobic in nature; (b) concentration-dependent; and (c) has an impact on the rheological properties of dispersion. PMID- 10387951 TI - Analysis of carboxyl content in oxidized celluloses by solid-state 13C CP/MAS NMR spectroscopy. AB - A noninvasive method to determine the carboxyl content in oxidized celluloses, using solid-state carbon-13 cross-polarization-magic angle spinning nuclear magnetic resonance (13C CP/MAS NMR) spectroscopy, has been developed. Standard samples containing 0, 4, 8, 12, 16, and 20% carboxyl content were prepared by mixing appropriate amounts of powdered cellulose, a non-oxidized cellulose standard prepared from cotton linter by ball-milling for 96 h, and a commercial oxidized cellulose sample that had a 20% carboxyl content. Standard curves were constructed by plotting the percent carboxyl content against the peak area at 171 ppm, normalized with (i) a peak area at 104 ppm and (ii) the sum of peak areas at 171, 62, and 65 ppm. The regression analysis of the curves yielded a linear relationship with correlation coefficient (R2) values of 0.9868 and 0.9863, respectively. To validate the methods, five new samples of oxidized cellulose were prepared and analyzed. The values obtained were comparable to those determined by the calcium acetate method described in the United States Pharmacopoeia (USP), indicating that the solid-state 13C CP/MAS NMR can be used to analyze carboxyl content in oxidized celluloses. PMID- 10387952 TI - Surfactant-induced leakage from liposomes: a comparison among different lecithin vesicles. AB - The interactions, at sublytic concentration, of Triton X-100 and sodium cholate with sonicated and extruded liposomes of egg and soya lecithins were considered to analyze the integrity and/or the barrier efficiency of liposomal membranes. Results are discussed in terms of surfactant partition between the aqueous and the lipid phases and of the release of a fluorescent hydrophilic probe. Phospholipid nature and liposome size influence detergent partition, whereas the content release is mainly affected by the surfactant mole fraction in the bilayer, and by the liposome size. PMID- 10387953 TI - Effect of a new chemoprotective agent, 2-(allylthio)pyrazine, on the pharmacokinetics of intravenous theophylline in rats. AB - The effect of 2-(allylthio)pyrazine (2-AP) pretreatment on the pharmacokinetics of theophylline and its metabolites was investigated after 1-min intravenous administration of aminophylline, 5 mg/kg as theophylline, to rats pretreated with three consecutive daily oral administration of 2-AP, 100 mg/kg. The AUC0-2 h of a metabolite of theophylline, 1,3-dimethyluric acid, 1,3-DMU (62.3 versus 106 microg/min per ml), and the percentages of intravenous dose of theophylline excreted in 24-h urine as 1,3-DMU (12.4% versus 20.8%, expressed in terms of theophylline) decreased significantly in 2-AP-pretreated rats when compared with those in control rats. Since CYP1A2 and CYP2E1 are involved in the formation of 1,3-DMU from theophylline, and 2-AP considerably suppressed CYP2E1 and tended to suppress CYP1A2 in rats, decreased formation of 1,3-DMU in 2-AP-pretreated rats could be mainly due to suppression of the CYP2E1 expression by pretreatment with 2-AP. PMID- 10387954 TI - Importance of the test medium for the release kinetics of a somatostatin analogue from poly(D,L-lactide-co-glycolide) microspheres. AB - The determination of in vitro release kinetics of peptides from poly(d,l-lactide co-glycolide) (PLGA) microspheres generally requires optimization of the test conditions for a given formulation. This is particularly important when in vitro/in vivo correlation should be determined. Here, the somatostatin analogue vapreotide pamoate, an octapeptide, was microencapsulated into PLGA 50:50 by spray-drying. The solubility of this peptide and its in vitro release kinetics from the microspheres were studied in various test media. The solubility of vapreotide pamoate was approximately 20-40 microg/ml in 67 mM phosphate buffer saline (PBS) at pH 7.4, but increased to approximately 500-1000 microg/ml at a pH of 3.5. At low pH, the solubility increased with the buffer concentration (1-66 mM). Very importantly, proteins (aqueous bovine serum albumin (BSA) solution or human serum) appeared to solubilize the peptide pamoate, resulting in solubilities ranging from 900 to 6100 microg/ml. The release rate was also greatly affected by the medium composition. Typically, in PBS of pH 7.4, only 33+/-1% of the peptide were released within 4 days, whereas 53+/-2 and 61+/-0.9% were released in 1% BSA solution and serum, respectively. The type of medium was found critical for the estimation of the in vivo release. The in vivo release kinetics of vapreotide pamoate from PLGA microspheres following administration to rats were qualitatively in good agreement with those obtained in vitro using serum as release medium. Finally, sterilization by gamma-irradiation had only a minor effect on the in vivo pharmacokinetics. PMID- 10387955 TI - Organic acids as excipients in matrix granules for colon-specific drug delivery. AB - Interest exists in developing site-specific systems for release of a drug in the lower part of the small intestine or in the colon. The aim of this study was to investigate whether drug release rates from enteric matrix granules could be influenced by using organic acids as excipients. Ibuprofen was used as a model drug and Eudragit S and Aqoat AS-HF as enteric polymers. The dissolution rates of the drug were investigated at different levels of pH (5.8, 6.8 and 7. 4). Drug absorption was studied in bioavailability tests in healthy volunteers. In vitro/in vivo correlation was also investigated. It was concluded that although inclusion of an organic acid in a formulation retarded in vitro release of the model drug, no corresponding effect was evident in in vivo studies. Bioavailability tests are therefore important early on during development of new dosage forms or formulations. Although no correlation between in vitro and in vivo results was generally evident correlation could be demonstrated for individual formulations following mathematical transformation of data. PMID- 10387956 TI - Anogenital human papilloma virus and the problem of persistence. AB - It has been known since antiquity that genital warts (condylomata acuminata) are a sexually transmitted disease. Since the late 1970s it has become increasingly clear that infections by the same virus, human papilloma virus (HPV) are closely implicated in the aetiology of anogenital squamous cell carcinomas and in their precursors, known as dysplasia, intraepithelial neoplasia (CIN) or squamous intraepithelial lesions (SIL). The natural history of these cancer precursors is characterised by regression, persistence or progression [1]. This review examines the natural history of anogenital HPV infections, especially persistence of the virus. Other topics covered include viral transmission, pathogenesis, the natural history of the lesions and treatment strategies. PMID- 10387957 TI - The immunology of genital human papilloma virus infection. AB - This paper reports a presentation by Margaret Stanley, Reader in Epithelial Biology, at the University of Cambridge, in which she reviews the evidence to date regarding the immunology of human papilloma virus (HPV) infection in genital warts. In this she explains that investigations into the immunology of genital wart infections indicate that the replication cycle of papilloma viruses is tightly linked to keratinocyte differentiation - a strategy for immune evasion. While the papilloma virus infects primitive basal cells, viral replication and viral assembly are confined to differentiating superficial epithelial cells. Viral replication and release are confined to cells destined for death and are not associated with inflammation. Such findings suggest that the immune system is ignorant or indifferent to the infection. Evidence from regressing genital warts in humans and animal models suggests that HPV is a cell-mediated immune response of the Th1 type offering a strategy for immunotherapy in benign disease. This is supported by evidence from trials with immunomodulatory agents. While strategies to elicit cytotoxic responses are required for malignant HPV associated lesions, the problems of immune evasion associated with these approaches should not be underestimated. Present optimal therapeutic strategies for genital human papilloma viruses infection would therefore appear to require the induction of a virus specific immune response, either by immunomodulatory agents and/or immunisation with the relevant viral antigens. PMID- 10387958 TI - Cytokine induction and modifying the immune response to human papilloma virus with imiquimod. AB - Imiquimod is the newest in a class of drugs known as immune response modifiers. In preclinical studies, imiquimod induced the production of cytokines - the principal cytokine for antiviral activity being interferon alpha. Imiquimod does not inhibit viruses directly, nor does it cause direct, non-specific cytolytic destruction [1]. Preclinical studies suggest that its antiviral action results from in vivo cytokine-induced activation of the immune system. This paper reviews a recent double blind, placebo-controlled study designed to evaluate this hypothesis. The results of this study showed that wart regression following treatment with imiquimod strongly correlated with a decrease in viral DNA and gene transcripts; a decrease in mRNA expression for proteins associated with cellular proliferation, and an increase in keratinocyte markers. These results support the hypothesis that stimulation of local cytokines by imiquimod leads to a reduction in human papilloma virus (HPV) load; to wart regression and to the normalisation of keratinocyte proliferation without evidence of scarring. PMID- 10387959 TI - Imiqimod in clinical practice. AB - Imiquimod 5% cream is a new compound which modifies the immune response by stimulating the production of interferon alpha and other cytokines. It has shown remarkable promise in the treatment of external genital and perianal warts when applied overnight, three times a week. It is also associated with a lower recurrence than those found with other current treatments. This paper reviews two of the pivotal multi-centre studies which confirm its efficacy in human papilloma virus (HPV) infections. These studies compared the effectiveness and safety of imiquimod 1% cream, imiquimod 5% cream and vehicle cream in the treatment of external anogenital warts. PMID- 10387960 TI - Questions from the audience AB - As part of the "Recurring Genital Warts" seminar at 6th Congress of European Academy of Dermatology and Venereology, audience members had the opportunity to question the speakers further about their clinical research. Issues raised focused mainly on the clinical efficacy, side effects and recurrence of imiquimod as an immune modifier, and about its potential future application. The questions and answers from this session are presented here. PMID- 10387961 TI - Perspectives on mitral-valve prolapse. PMID- 10387962 TI - We've Got a Treatment, but What's the Disease? Or A Brief History of Hypofractionation and Its Relationship to Stereotactic Radiosurgery. AB - Hypofractionation has been a recurring issue during the near century-long history of radiation oncology. Coutard first introduced protracted dose-fraction regimens that uniquely allowed for the control of "deep" tumors. Subsequent studies have consistently shown that hypofractionation leads to an increase in complication rates and a paradoxical decrease in cure rates. There have, nonetheless, been several resurgences of interest in hypofractionation, based on titration of treatment for acute isoeffects and for the accommodation of an adjuvant treatment relating to presumed hypoxia-induced resistance, and more recently, stereotactic radiosurgery. In final analysis of the earlier studies, the same effects on cure and complication were noted and there was a return to multi-fractionation. Stereotactic radiosurgery is now being evaluated. Stereotactic radiosurgery takes hypofractionation to an extreme by use of a single, large fraction of radiation therapy. In doing so, the late effects radiation oncologists ordinarily strive to avoid are brought about intentionally, minimizing or even eliminating any therapeutic index within the treatment volume. Stereotactic radiosurgery has been used successfully for treatment of benign lesions such as arteriovenous malformations in which total volume necrosis of small dimensions appears to be efficacious therapy. Stereotactic radiosurgery has also recently been extrapolated to malignant tumors in the brain which require larger treatment volumes, but the data on outcome following such treatment remain sparse. Therapeutic index must be preserved to obviate an intolerable volume of necrosis and other late effects. The single fraction approach to stereotactic radiosurgery for cancer is vulnerable to the same radiobiological criticisms that have been a recurrent theme with hypofractionation. Fractionated stereotactic radiosurgery is far more consistent with the principles of conventional radiobiology and oncology and represents the quintessential application of three-dimensional treatment planning. Stereotactic radiosurgery is really stereotactic radiotherapy, and when applied in single fraction to the treatment of cancer, it is suboptimal radiation oncology. Its utilization is virtually predicated on the ability to perform another craniotomy to remove focal necrosis. PMID- 10387963 TI - Locally Advanced Breast Cancer. AB - Locally advanced breast cancer encompasses a heterogeneous collection of breast neoplasms and constitutes approximately 10%-20% of the newly diagnosed breast cancers. These cancers may have widely different clinical and biological characteristics. Patients with these tumors may be classified as stage IIB, III or IV breast cancer according to the American Joint Committee for Cancer Staging and End Results Reporting (TNM classification). Multidisciplinary therapy has become the treatment of choice for these patients. Primary or neoadjuvant chemotherapy followed by locoregional therapy, either surgery and/or radiotherapy, and postoperative systemic chemotherapy is now an accepted strategy. More than 70% of patients achieve an objective response (including pathological complete remission in 10%-25% of cases), and many patients experience downstaging through primary chemotherapy. Breast conservation is possible in 10%-40% of patients with locally advanced breast cancer; almost all patients initially are rendered disease-free, and long-term local control is achieved in over 70% of these patients. Primary chemotherapy is the initial choice of treatment for patients with locally advanced tumors, but it is unclear what the optimal sequence of subsequent therapies should be, whether one or two local treatment modalities are necessary, and whether any or different postoperative chemotherapy is needed. The efficacy of primary chemotherapy was demonstrated in several large prospective studies in patients with locally advanced breast cancer. The natural history of this disease was changed dramatically by the introduction of these combined modality therapies. Five-year disease-free survival rates of 35%-70% are commonly reported, and about 25%-40% of patients will survive beyond 10 years without recurrence. In summary, multidisciplinary therapy that includes primary chemotherapy provides appropriate local control and the possibility of breast conservation therapy; it increases surgical resectability and survival rates in patients with locally advanced breast cancer. The role of new innovative therapeutic strategies such as high dose chemotherapy, with hematopoietic stem cell rescue, new cytotoxic agents and higher dose-intensity therapy is currently under evaluation in patients with locally advanced breast cancer. PMID- 10387964 TI - Intraperitoneal Therapy of Ovarian Cancer. AB - PURPOSE: This is a review of the rationale for the intraperitoneal administration of antineoplastic agents in the management of ovarian cancer. PATIENTS AND METHODS: Patients have been treated in a number of clinical trials to define the toxicity profile and efficacy of intraperitoneal therapy in this clinical setting. RESULTS: Phase I-II trials have confirmed that a number of cytotoxic and biological agents can be administered into the peritoneal cavity as treatment of ovarian cancer with an acceptable toxicity profile and with the attainment of surgically documented responses (including complete responses). In addition, a recently reported phase III trial comparing initial treatment of small-volume residual advanced ovarian cancer with either intravenous or intraperitoneal cisplatin concluded that the intraperitoneal route of drug administration was associated with a longer survival and less toxicity. DISCUSSION: In the salvage (second-line) setting, responses to intraperitoneally administered antineoplastic agents are seen principally in individuals with small-volume residual disease (largest tumor mass /= 6 percentage points, compared to corresponding quality-of-life changes in placebo-treated patients. rHuEPO was well tolerated compared to placebo. The above results suggest that rHuEPO may be a useful agent to palliate the morbid consequences of the anemia that is often found in association with advanced cancer. PMID- 10387981 TI - What Does Multidrug Resistance (MDR) Expression Mean in the Clinic? AB - For 15 years, an overflowing literature has been published about MDR-1 gene expression in tumor cell lines and in cancerous tissues at various stages of disease and treatment (chemotherapy-naive, during treatment and at relapse). However, the clinical significance of this particular feature, if it seemed obvious in the 1980s as a factor responsible for the development of chemoresistance, is currently reconsidered. MDR-1 gene expression seems to be, at least in some instances, a hallmark of tumor cell aggressiveness and of chemoresistance rather than its cause, the mechanisms of which are probably far more complex. The failure of MDR reversal trials might result from the misunderstood or overvalued role of MDR expression in cancer cells rather than from a lack of control of pharmacological parameters. This review summarizes recent data and hypothesis about the expression of P-170 and its clinical significance in some important human tumor types, suggesting that it should rather be considered in the future as an adverse prognostic factor. PMID- 10387982 TI - Bone Marrow Transplantation for Cancer - An Update. AB - The number of allogeneic and autologous bone marrow transplants continues to grow worldwide. Bone marrow transplantation (BMT) has become standard therapy for many patients with leukemia, lymphoma, multiple myeloma and testicular cancer. Encouraging results of autologous BMT in treating patients with poor-risk breast cancer have led to this approach being tested in nationwide randomized trials. In order to increase availability and efficacy of BMT, other sources of hematopoietic cells are explored for transplantation, such as from HLA-matched unrelated volunteer donors, partially matched related donors, placental/umbilical cord blood and allogeneic peripheral blood. Relapse of original malignancy remains the main obstacle for the success of BMT. Recent clinical investigations have demonstrated that donor-derived peripheral blood leukocytes are effective in inducing remissions in patients with hematological malignancies who relapse after allogeneic BMT. BMT procedures are associated with significant complexity and should be carried out only in transplant units that meet adequate standards. In order to better define the role of BMT in treating cancer, more phase III clinical trials are needed. The future of BMT will depend on further improvements in its efficacy and economic constraints. PMID- 10387983 TI - Pediatric Clinical Trials. AB - Clinical trials specifically tailored to the unique tumors and leukemias of children have resulted in increased survival rates approaching or exceeding 70% for most diseases. These studies have been carried out by investigators at large, independent institutions or through the auspices of the Children's Cancer Group or Pediatric Oncology Group. The National Cancer Institute has also supported pediatric disease-focused efforts for rhabdomyosarcoma, Wilms tumor, Ewing's sarcoma, osteosarcomas and other diseases. The present-day training of pediatric hematology/oncology fellows assures continuing contributions to the biology of childhood malignant lesions through applications of translational research. PMID- 10387984 TI - Alternative and Complementary Cancer Treatments. AB - Alternative and complementary therapies differ importantly, and the distinction between the two is crucial for clinical oncologists. "Alternative" or unproven therapies are treatments used independent of surgery, radiation and chemotherapy. They can be dangerous directly and also by delaying patients' receipt of mainstream care. In contrast, complementary therapies typically are adjuncts to mainstream medicine. They can provide symptom control and noninvasive palliation with minimal side effects, improve patients' well-being and enhance cancer medicine. Complementary therapies represent a desired addition and balance to technologically sophisticated cancer care. PMID- 10387985 TI - Anticancer Drug Development: The Way Forward. AB - Cancer chemotherapy celebrated its fiftieth anniversary last year. It was in 1945 that wartime research on the nitrogen mustards, which uncovered their potential use in the treatment of leukaemias and other cancers, was first made public. Fifty years later, more than sixty drugs have been registered in the USA for the treatment of cancer, but there are still lessons to be learnt. One problem, paradoxically, is that many anticancer agents produce a response in several different classes of the disease. This means that once a new agent has been shown to be effective in one cancer, much effort is devoted to further investigations of the same drug in various combinations for different disorders. While this approach has led to advances in the treatment of many childhood cancers and some rare diseases, a plethora of studies on metastatic colon cancer, for example, has yielded little benefit. 5-fluorouracil continues to be used in trials, yet there is no evidence for an increase in survival. The lesson to be learnt is that many common cancers are not adequately treated by present-day chemotherapy, and most trials of this sort are a waste of time. Significant increases in survival will only occur if the selectivity of present-day anticancer agents can be increased or new classes of more selective agents can be discovered. There are two fundamental problems in drug development: a lack of suitable laboratory tests and the difficulty of conducting early clinical trials. Firstly, no existing laboratory method can accurately predict which chemical will be effective against a particular class of human cancer. At best, tests can demonstrate a general 'anticancer' property. This is well exemplified by the discovery of cisplatin. The fact that cisplatin caused regression in a number of transplanted rodent tumours created no great excitement amongst chemotherapists. It was only later when it was tested clinically against ovarian cancer that results were sufficiently positive to encourage others to investigate. Only then was it discovered that metastatic teratoma was extraordinarily sensitive to the drug. This finding was made as a result of phase II trials and no laboratory model could have predicted it. The lesson to be learnt is that new drugs should be tested extensively in phase II trials before they are discarded. The second problem concerns early clinical trials. Because new drugs can only be tested against advanced and usually heavily pretreated disease, it is unlikely that dramatic responses will occur. The methods used to detect responses in solid tumours and metastases are crude, and it is likely that many useful drugs are missed. New techniques are needed to detect small but important responses. In addition to these technical problems, clinical trials are expensive and the time required for preclinical pharmacology and toxicology is lengthy. In the early days, drugs could enter clinical trials after fairly simple toxicological studies. The thalidomide disaster in the 1960s, however, led to the setting up of regulatory bodies to scrutinize drugs before clinical trials. This proved detrimental for cancer drug development because a series of fairly long-term tests is now required. These must be carried out in both rodents and one other species, usually the dog. This approach was probably a good thing for most medicines where a large margin of safety is required between the therapeutic dose and the dose which causes side effects, but was inappropriate for anticancer agents which are tested at the maximum possible dose which gives manageable side effects. These new regulations meant that the cost of one clinical trial after the 1970s was equivalent to the cost of ten before that time. Solutions to these problems are available, although to put them into practice would require the cooperation of government regulatory authorities, the pharmaceutical industry and other organisations such as the US National Cancer Institute (NCI), the UK Cancer Research Campaign (CRC) and the European Organisation for Research and Treatment of Cancer (EORTC). Firstly, it is important to switch from clinical trials of analogues and combinations of standard drugs to trials of new classes of anticancer agents. Further, an international effort should be launched whereby these new agents can be rapidly tested in phase II trials against common solid cancers using new techniques to detect small but significant tumour responses. Lead chemicals discovered in this way could then be taken back to the laboratory for further development. There is no shortage of new drugs which act by mechanisms quite different from present-day agents, and new approaches can greatly increase the amount of cytotoxic agents delivered to solid tumours. As long ago as 1980, the CRC introduced protocols which enabled early clinical trials to be carried out rapidly and with minimal cost. These procedures used short-term tests only in rodents to determine a safe starting dose. The test can be completed within six months and around fifty clinical trials using this protocol have been successfully carried out in collaboration with the EORTC. Despite this, the American Food and Drug Administration (FDA), regulatory authorities in many other countries and many drug companies still insist on using a second animal species before a phase I clinical trial is permitted instead of using the money spent to develop several agents with minimal toxicology testing. The EORTC and CRC also plan to introduce positron emission tomographic scanning into early clinical trials as a highly sensitive method of measuring tumour response. Cancer mortality has changed little over the past forty years, mainly because of our failure to develop curative chemotherapy for the common solid cancers. The way forward is to carry out extensive phase I and II clinical trials of the many new types of anticancer agent that have become available as a result of increased knowledge about cancer cells and how they differ from normal tissues. In order to do this, the regulatory authorities must recognize that minimal toxicology protocols are adequate, and drug companies must be persuaded to give more emphasis to the search for new chemotherapeutic agents. A coordinated effort to achieve these aims would be a wonderful way to mark the fiftieth anniversary of modern chemotherapy. Unfortunately the regulatory authorities find it less risky to stick with extensive safety testing rather than to use shortcuts, however well-validated clinically. Many but not all drug companies, mindful of profits, prefer the easy way out and concentrate on analogues, while most clinicians opt for trials of combinations of known agents, being aware that they are worth a publication or two. Reprinted with permission from Helix, Volume V, Issue 1, 1996, pp. 20-21. PMID- 10387986 TI - Special Education: Aplastic Anemia. AB - WHAT IS HYPOPLASTIC ANEMIA? Aplastic anemia is a hematological disease characterized by pancytopenia and bone marrow hypoplasia. Acquired cases of aplastic anemia are almost all idiopathic and arise from unknown causes. Other cases of aplastic anemia are secondary and are caused by radiation, chemicals or viruses. PATHOPHYSIOLOGY: Aplastic anemia is manifested as a marked reduction in the number of pluripotent hematopoietic stem cells, but why this occurs is still uncertain. Some of the proposed causes include abnormalities of the hematopoietic stem cells, abnormalities in the hematopoietic microenvironment, and immunologically mediated damage to the hematopoietic stem cells (Figure 1). ABNORMALTIES OF THE HEMATOPOIETIC STEM CELLS: Patients with aplastic anemia, and long-term survivors in particular, are at increased risk of developing paroxysmal nocturnal hemoglobinuria (PNH), myelodysplastic syndrome (MDS), or acute myelocytic leukemia. This suggests that, in at least some of these patients, the hematopoietic stem cells themselves are abnormal. It also suggests that in some of these patients the blood cells are clonal (that is, all the blood cells are derived from a single pluripotent stem cell). In short, what these findings imply is that aplastic anemia may be caused by the emergence of an abnormal clone. Clonal hematopoiesis, however, can also be considered nothing more than a consequence. In other words, it is possible that hematopoiesis in this kind of patient is performed by a lone pluripotent stem cell that somehow managed to survive eradication. No definitive interpretation of clonal hematopoiesis has been agreed upon, and it is still a topic for future research. ABNORMAL HEMATOPOIETIC MICROENVIRONMENT: The presence of stromal cells, which form the microenvironment of bone marrow, is very important in hematopoiesis. Hematopoietic stem cells proliferate and differentiate either by adhering to stromal cells or by being stimulated by the various hematopoietic factors that stromal cells produce. Therefore, it is quite possible that aplastic anemia is caused by abnormalities in the hematopoietic microenvironment. However, many separate studies have demonstrated that the hematopoietic microenvironment in the vast majority of aplastic anemia cases is normal. IMMUNE MECHANISMS: Immunosuppressive agents are often effective in treating aplastic anemia, and therefore it is believed that immunological mechanisms contribute to the disease in more than half the cases. The following mechanisms have been proposed as causes for the onset of immunologically mediated aplastic anemia: * Decreases in Hematopoietic Factors Produced by Monocytes and Lymphocytes. Some patients with aplastic anemia show decreased production of interleukin 1 (IL-1) by peripheral blood monocytes, and it is possible that a drop in the concentration of this factor is linked to the onset of the disease [1]. It is also possible, however, that decreased IL-1 production by monocytes is not a cause of the disease, but merely a consequence. Moreover, no cases have been reported that exhibit reduced production of hematopoietic factors produced by lymphocytes such as GM-CSF, IL-3, or IL-6. * Damage by Cytokines that Suppress Hematopoiesis. It has been reported that increased levels of interferon &ggr; (IFN-&ggr;), which is produced by lymphocytes, and tumor necrosis factor &agr; (TNF-&agr;), which is produced by monocytes and macrophages, are found in the bone marrow and peripheral blood of aplastic anemia patients [2, 3]. These two factors act as suppressors of hematopoiesis, and it is possible that they contribute to the disease. The increase of these inflammatory cytokines in the bone marrow strongly suggests the presence of either specific or non-specific destruction of the hematopoietic stem cells by immunoregulatory cells. * Suppression of Hematopoiesis by Cytotoxic T Cells (Killer T Cells). Cases have been reported in which cytotoxic T cell clones that damage the autologous hematopoietic precursor cells are present [4]. Therefore, we can easily conceive of a mechanism in which these cytotoxic T cells specifically destroy the hematopoietic stem cells and cause aplastic anemia. * Suppression of Hematopoiesis by Natural Killer (NK) Cells. NK activity of aplastic anemia patients is depressed, and, generally speaking, it is highly unlikely that NK cells contribute to this condition. However, it has been reported that clonal NK cells are thought to cause the disease in patients exhibiting pancytopenia and bone marrow hypoplasia. Therefore, when this disease is diagnosed, a peripheral blood granular lymphocyte count and NK cell surface marker analysis should always be performed. DIAGNOSIS: A necessary condition for the diagnosis of aplastic anemia is the presence of pancytopenia. Moreover, it is necessary to rule out all other causes of pancytopenia. It is especially important in differential diagnosis to look for PNH and MDS. In cases of aplastic anemia there are patients that exhibit PNH during the course of the disease, and this condition is called aplastic anemia-PNH syndrome. It has recently been shown that bone marrow and peripheral blood cells in some patients diagnosed with aplastic anemia are partially lacking GPI anchor proteins (CD16, CD55, and CD59) [5]. Whether such patients become to exhibit aplastic anemia-PNH syndrome in the future remains to be elucidated. In MDS the bone marrow generally exhibits normoplasia or hyperplasia, and only in rare cases does it exhibit hypoplasia. This condition is referred to as hypoplastic MDS. Hypoplastic MDS can be differentiated from aplastic anemia by the presence of abnormal cell morphology that is sometimes accompanied by chromosomal abnormalities. TREATMENT:Aplastic anemia is treated with androgens, high-dose methylprednisolone, cyclosporin A (CyA), antithymocyte globulin (ATG), antilymphocyte globulin (ALG), hematopoietic growth factors such as G-CSF, and bone marrow transplantation. Interestingly, patients who require continuous CyA administration to maintain stable hematopoiesis have a specific HLA class II haplotype (DRB1*1501-DQA1*0102 DQB1*0602) [6]. Recent reports from EBMT SAA Working Party showed the excellent therapeutic result (response rate 82%) when severe cases were treated with ALG, CyA and G-CSF in combination [7]. PMID- 10387987 TI - Cancer Drug Development: New Targets for Cancer Treatment. AB - There is often a considerable lapse of time between the definition of what causes a disease in the laboratory and the development of successful therapy. However, the history of medicine teaches us that the need to understand the scientific basis of disease before the discovery of new treatments is both essential and inevitable. During the middle of the 19th century, the work of the great German pathologist, Rudolf Virchow, defined disease as having an anatomic or histologic basis. In the clinic, this scientific perspective would lead to increasingly effective and, often, increasingly aggressive surgical approaches to disease. Later in the 19th century, Koch's discovery of the tubercle bacillus (a discovery Virchow disbelieved and publication of which he thwarted, since he hypothesized that cancer, not microbes, caused consumption!), would define a microbiological basis for disease. With bacteria defined as a major cause of human suffering, the stage was set for the development of the discovery of effective antibiotics. In the early 20th century, the pioneering work of Banting, Best and others would show that disease can also have an endocrine or metabolic basis. This new body of scientific knowledge would lead not only to the specific discovery of insulin as an effective treatment for diabetes but also to a more general understanding of the role of hormones, vitamins and co-factors in human health and disease. Basic medical research and its successful translation into effective treatments has fundamentally altered the cause of human death. In the developed world, where access to the benefit of this work is available, infectious disease is not the problem it was in the days of Pasteur, Metchnikoff and Ehrlich. As we approach the millennium, science is now teaching us that diseases, particularly cancer, can have a molecular or genetic basis. Can successful application of this new knowledge be far behind? We are already seeing the application of this new knowledge in cancer drug screening and cancer drug development. At the NCI, for example, the old in vivo mouse screen using mouse lymphomas has been shelved; it discovered compounds with some activity in lymphomas, but not the common solid tumors of adulthood. It has been replaced with an initial in vitro screen of some sixty cell lines, representing the common solid tumors-ovary, G.I., lung, breast, CNS, melanoma and others. The idea was to not only discover new drugs with specific anti-tumor activity but also to use the small volumes required for in vitro screening as a medium to screen for new natural product compounds, one of the richest sources of effective chemotherapy. The cell line project had an unexpected dividend. The pattern of sensitivity in the panel predicted the mechanism of action of unknown compounds. An antifolate suppressed cell growth of the different lines like other antifolates, anti-tubulin compounds suppressed like other anti-tubulins, and so on. It now became possible, at a very early stage of cancer drug screening, to select for drugs with unknown-and potentially novel-mechanisms of action. The idea was taken to the next logical step, and that was to characterize the entire panel for important molecular properties of human malignancy: mutations in the tumor suppressor gene p53, expression of important oncogenes like ras or myc, the gp170 gene which confers multiple drug resistance, protein-specific kinases, and others. It now became possible to use the cell line panel as a tool to detect new drugs which targeted a specific genetic property of the tumor cell. Researchers can now ask whether a given drug is likely to inhibit multiple drug resistance or kill cells which over-express specific oncogenes at the earliest phase of drug discovery. In this issue of The Oncologist, Tom Connors celebrates the fiftieth anniversary of cancer chemotherapy. His focus is on the importance of international collaboration in clinical trials and the negative impact of unnecessary bureaucracy and regulation. As a student of Tom's in the 1970s in London, working on hepatoma-specific alkylating agents at Charing Cross Hospital in collaboration with his lab on the other side of town, I can attest to the fact that the regulatory hurdles to cancer drug development just twenty years later have added immeasurably to the effort and cost of cancer drug development. However, I look with optimism to the future of cancer diagnosis, prevention and treatment. It is a future where what we are learning now about the molecular and genetic basis of cancer will find their clinical outlet just as surely as the anatomic, microbial, metabolic and endocrine basis for disease has in the past. This new knowledge will provide new techniques in molecular diagnosis, which will allow us to predict which in situ cancers are destined for malignant behavior, and which can be safely watched without the need for intervention. Individual patient risk for particular cancers will be accurately predictable, so that patients can alter lifestyle habits or begin other prevention strategies. Oncogenes and growth suppressor genes give us new targets to inhibit or replace. Tumor-specific kinases will meet their inhibitors. The oncologist will play a leading role in understanding, applying and interpreting this new information in the clinic-an exciting and challenging future! PMID- 10387988 TI - Interleukin 1 Trials in Cancer Patients: A Review of the Toxicity, Antitumor and Hematopoietic Effects. AB - Clinical trials of interleukin 1&agr; (IL-1&agr;) and IL-1&bgr; have been completed that assess the toxicities of these cytokines as well as their hematopoietic and antitumor effects. Both forms of IL-1 recognize the same cell surface receptors and have similar toxicities and similar biological activities. Toxicities including fever, flu-like symptoms and dose-limiting hypotension can be severe yet manageable, and IL-1 can be given safely to human cancer patients. Most toxicities and biological effects appear to be dose-related. IL-1 alone has little antitumor activity against melanoma, renal cell carcinomas or other malignancies. The hematopoietic effects, including megakaryocytopoietic effects, are modest and are probably not worth the toxicity necessary to achieve them. However, IL-1 seems to endow certain progenitor cells with responsiveness to other hematopoietic cytokines including colony-stimulating factors and IL-3. One potential application of IL-1 is to help expand bone marrow ex vivo following stem cell harvest, which could allow further chemotherapy dose escalations in chemotherapy-sensitive tumors. PMID- 10387989 TI - Diagnosis and Treatment of Early-Stage Non-Small Cell Lung Cancer. AB - Current recommendations for the diagnostic work-up and treatment of early-stage non-small cell lung cancer are presented, and the rationale behind these recommendations is reviewed. Early-stage disease is found in approximately 30% of patients at initial presentation. Surgeons continue to be uncertain with regard to how extensively they should look for metastatic disease, especially in asymptomatic patients with newly diagnosed lung cancer. While it is generally agreed that surgery is an important component of treatment for stage I and II non small cell lung cancer, the role of adjuvant therapies in early-stage disease merits further study. Stage IIIa lung cancer is evolving as a disease for which multimodality therapy is likely to play a role, but the timing and sequence of treatment is an area of intense investigation. The recommendations made in this article are based upon the results of randomized clinical trials whenever possible. PMID- 10387990 TI - Multimodality Therapy for Esophageal Cancer. AB - Adjuvant and neoadjuvant therapeutic principles have in recent years received increasing attention in the management of patients with esophageal cancer. A series of randomized prospective trials has convincingly demonstrated that adjuvant postoperative radiation or chemotherapy does not result in a survival advantage after a complete tumor resection. The available data on the role of neoadjuvant preoperative therapy in patients with adenocarcinoma or squamous cell carcinoma of the esophagus are not yet conclusive. While neoadjuvant therapy may undoubtedly reduce the tumor mass in a substantial portion of patients, a series of randomized controlled trials has shown that compared with primary resection, a multimodal approach does not result in a survival benefit in patients with locoregional, i.e., potentially resectable, tumors. In contrast, in patients with locally advanced tumors, i.e., tumors in which a complete tumor removal with primary surgery appears unlikely, neoadjuvant therapy allows a marked downstaging of the primary tumor and thus significantly increases the chance for complete tumor removal on subsequent surgery. However, only patients with objective clinical or histopathological response to preoperative therapy appear to benefit from this approach. Compared with preoperative chemotherapy alone, combined radiochemotherapy increases the rate of response but may also increase postoperative morbidity and mortality. Neoadjuvant therapy should therefore currently be performed only in experienced centers within the context of clinical trials. The identification of factors which would facilitate prediction of the response to neoadjuvant therapy is currently the focus of several studies. Furthermore, more effective and less toxic preoperative therapy regimens are required to increase the response rates and combat systemic recurrences. PMID- 10387991 TI - A Tribute to Bruce Chabner. PMID- 10387992 TI - Resistance Mechanisms to Methotrexate in Tumors. AB - The mechanisms of intrinsic and acquired resistance to methotrexate (MTX) in human tumors are reviewed herein. In blasts from patients with acute lymphocytic leukemia, resistance mechanisms found are decreased uptake and increased dihydrofolate reductase (DHFR) activity. A major cause of intrinsic resistance to MTX in soft tissue sarcoma cells and in acute myelocytic leukemia appears to be a lack of drug retention, due mainly to low levels of polyglutamylation. A novel association between lack of the retinoblastoma protein and intrinsic MTX resistance has been found. This has been attributed to an increase in DHFR activity, due to an increased rate of transcription of this gene, stimulated by an increase in levels of free E2F, not sequestered by hypophosphorylated retinoblastoma protein. PMID- 10387993 TI - Modulating the Radiation Response. AB - This review describes some current and future approaches designed to modulate the response of tumors and normal tissues to cell killing by ionizing radiation. The emerging knowledge of tumor, cellular and molecular biology is providing a better understanding of the clinical results with the hypoxic cell sensitizers and novel approaches to radiation sensitization and protection. PMID- 10387994 TI - From Plasma Kinetics to Cellular Pharmacology. AB - This paper primarily summarizes the work done under the guidance of Dr. Bruce Chabner in 1975 and 1976. During these years I studied the role of drug concentration, duration of exposure, and endogenous metabolites in determining methotrexate (MTX) cytotoxicity to the bone marrow stem cell (CFU-C). We found that the rate of loss of CFU-C during continuous intravenous infusion of methotrexate was related to the duration of exposure until the nadir in cell count was reached at 24 hours. Depletion of nucleated cells was mitigated, probably as a result of recruitment of previously uncommitted precursor cells to CFU-C, even while the MTX infusion was continued up to 48 hours. In vitro it was shown that methotrexate and leucovorin were transported competitively in the CFU C, which was in clear contrast with rescue agents such as thymidine and nucleoside analogs. After my training my work has continued in close contact with Dr. Chabner, while the scope of both our interests broadened to include the field of multidrug resistance (MDR). My own interest focused on functional studies in MDR. PMID- 10387995 TI - The Importance of Drug Scheduling in Cancer Chemotherapy: Etoposide as an Example. AB - Etoposide is a drug whose antineoplastic activity is dependent on the schedule of drug administration. This article reviews the rationale for a prolonged schedule of etoposide administration and the therapeutic results of use of such a schedule in the treatment of cancer. The pharmacology of etoposide is also reviewed, with particular attention paid to the pharmacokinetics of oral etoposide and etoposide plasma concentrations associated with cytotoxicity. PMID- 10387996 TI - The Clinical Development of Paclitaxel: A Successful Collaboration of Academia, Industry and the National Cancer Institute. AB - The successful development of paclitaxel as an important new antineoplastic agent with the potential to have an impact on a number of human cancers was possible as a result of significant contributions from individuals and groups with diverse areas of interest and expertise. The advancement of paclitaxel through the preclinical and clinical evaluation which ultimately led to its approval, as well as surmounting the regulatory hurdles which were faced required the close collaboration of individual investigators at academic institutions, the pharmaceutical industry (Bristol-Myers Squibb) and the National Cancer Institute. The latter stages of this developmental effort can be viewed as a prime example of the potential of the Cooperative Research and Development Agreement mechanism to bring novel therapies to patients with serious illnesses in a timely fashion. It is also tangible evidence of the vision and perseverance of a number of members of the Division of Cancer Treatment under the direction of Dr. Bruce Chabner, in whose honor this symposium is given. PMID- 10387997 TI - Methotrexate: An Effective Agent for Treating Cancer and Building Careers. The Polyglutamate Era. AB - This paper chronicles developments in the laboratory of Dr. Bruce Chabner during the period 1978-1981. Initial work demonstrated that methotrexate is taken up by human breast cancer cells by a high-affinity, carrier-mediated, energy-dependent transport system similar to that described in murine leukemia cells. Conversion of methotrexate to a high molecular weight polyglutamate metabolite was also demonstrated to occur in human breast cancer cells. Polyglutamates became the predominant form of intracellular drug, both free in the cytosol and bound to dihydrofolate reductase, during a 24 h exposure to clinically achievable methotrexate concentrations. Intracellular retention of polyglutamates led to prolonged suppression of thymidine synthesis and loss of cell viability after removal of extracellular drug. This work identified methotrexate polyglutamates as biologically active enzyme inhibitors in human tumor cells and launched a series of investigations on the interaction of these derivatives with folate requiring enzymes. PMID- 10387998 TI - Biochemical and Molecular Studies of Human Methenyltetrahydrofolate Synthetase. AB - 5-FormylH(4)folate is administered clinically under the name Leucovorintrade mark in association with the antineoplastic agent 5-fluorouracil (5-FU) to enhance the cytotoxic effects of 5-FU. The combination has been shown to be superior to 5-FU alone in the treatment of patients with metastatic colorectal carcinoma. Methenyltetrahydrofolate synthetase (MTHFS) catalyzes the transformation of 5 formyltetrahydrofolate to methenylH(4)folate, which is the obligatory initial metabolic step prior to the intracellular conversion of 5-formylH(4)folate to other reduced folates and the increase in intracellular folate pools required for 5-FU potentiation. In the following paper, we will summarize results of biochemical and molecular studies of human MTHFS. PMID- 10387999 TI - New Concepts for the Development and Use of Antifolates. AB - Approximately one-third of all cases of colorectal carcinoma present in an advanced and, therefore, incurable stage. For these patients, the development of new chemotherapeutic strategies is of central importance. Biochemical modulation of 5-fluorouracil (5-FU) has resulted in approximately a twofold increase in activity of 5-FU. Recent preclinical investigations suggest that interferon can also modulate the activity of 5-FU and may result in enhanced response rates in patients. One of the critical mechanisms of resistance to 5-FU appears to be the acute induction in thymidylate synthase (TS) levels following therapy with inhibitors of this enzyme. This mechanism is based on a novel autoregulatory feedback pathway wherein the TS protein regulates its own translational efficiency. Regulatory function of the enzyme is dependent on its state of occupancy by either the physiologic ligands or inhibitors, including fluoropyrimidines and antifolates. Ongoing efforts are directed toward utilizing knowledge of this protein/messenger RNA interaction for therapeutic benefit. Given the importance of TS, our laboratory has developed antibodies capable of quantitating the levels of this enzyme in fresh or paraffin-embedded tissues. Preliminary investigations suggest that the level of TS has prognostic importance in patients with rectal carcinoma and may be used to predict responsiveness to fluoropyrimidine agents. Novel strategies utilizing dual modulation of 5-FU with leucovorin and interferon are under investigation in both the advanced and adjuvant disease settings. Emerging mechanistic concepts regarding TS, along with the development of new, more potent inhibitors will hopefully result in future therapeutic gains. PMID- 10388000 TI - P-Glycoprotein, Multidrug Resistance and Protein Kinase C. AB - The multidrug resistant (MDR) phenotype is a well-studied subject that has been recognized as a determinant underlying specific types of drug resistance in human cancer. Although it is clear that the P-glycoprotein plays a major role in MDR, it is not clear whether post-translational modifications such as phosphorylation have any major impact on its modulation. The laboratory of Dr. Bruce Chabner was one of the first to describe increased expression and activity of protein kinase C (PKC) associated with the MDR phenotype. Since that time, a similar correlation has been observed in many other MDR cell lines. Most of these studies have been performed with doxorubicin-selected cells that have acquired MDR and have shown increased PKC activity, mainly for PKC-a isoenzyme. Intrinsic MDR in human renal cell carcinoma lines has been shown to correlate directly with PKC activity, but further studies with intrinsic MDR cell lines are needed before any conclusions can be drawn. More recent evidence suggests that there is a complex biochemical process by which PKC isoenzymes differentially phosphorylate specific serine residues in the linker region of P-glycoprotein which may lead to alterations in P-glycoprotein ATPase and drug-binding functions. To further complicate matters, PKC plays an important role in anti-apoptotic pathways, which can confound the dissection and elucidation of drug-resistance mechanisms. However, these areas are still under active investigation and not fully answered. Further studies are needed to specifically answer the question of whether PKC directly modulates basal and/or drug-stimulated P-glycoprotein function. This manuscript reviews the majority of the literature on PKC and MDR, as well as offers caveats for interpretation of these studies to answer the above questions. PMID- 10388001 TI - Clinical Reversal of Multidrug Resistance. AB - Reversal of drug resistance offers the hope of increasing the efficacy of conventional chemotherapy. We tested dexverapamil as a P-glycoprotein antagonist in combination with EPOCH chemotherapy in refractory non-Hodgkin's lymphoma. In a cross-over design, dexverapamil was added to EPOCH after disease stabilization or progression occurred. Objective responses were observed in 10 of 41 assessable patients. Biopsies for mdr-1 were obtained before EPOCH treatment and at the time of cross-over to dexverapamil. Levels of mdr-1 were low before EPOCH, but increased fourfold or more in 42% of patients in whom serial samples were obtained. Pharmacokinetic analysis revealed median peak concentrations of dexverapamil and its metabolite, nor-dexverapamil, of 1.66 umol/l and 1.58 umol/l, respectively. Since both are comparable antagonists, a median peak total reversing concentration of 3.24 umol/l was achieved. Pharmacokinetic analysis of doxorubicin and etoposide levels confirmed a delay in the clearance of doxorubicin ranging from 5% to 24%; no change in the pharmacokinetics of etoposide was observed. This study provides sufficient rationale for testing dexverapamil in a randomized clinical trial. PMID- 10388002 TI - Closing Remarks. AB - About twenty years ago, the leaders of the National Cancer Institute (NCI) decided to start a new branch in the Clinical Oncology Program of the Division of Cancer Treatment. That new entity was named the Clinical Pharmacology Branch (CPB), and its first leader was a brilliant, young, promising investigator named Bruce A. Chabner. Chabner was educated at Yale and Harvard, and appeared to have an extraordinary grasp of novel concepts that were being developed in the emerging area of cancer chemotherapy. What the NCI leaders did not fully appreciate at the time was that they had just given birth to one of the most extraordinary careers in academic medicine. From the early seventies through the early eighties, Bruce Chabner developed a strong laboratory program that was based on scientific discovery and on the development of new talent. The CPB focused on new drug development, elucidation of drug mechanism(s) of action, and the development of new ways to use drugs that were already available. Concurrent with this laboratory effort was active participation in the development of clinical treatment regimens for Hodgkin's disease, non-Hodgkin's lymphoma, and other malignancies. Individuals who trained under Chabner are now cancer center directors, department heads, laboratory chiefs, and hold many other high-profile positions. From 1981 to 1995 Bruce Chabner was Director of the Division of Cancer Treatment (DCT) of the NCI. In that capacity he was Scientific Director of the Intramural Program within DCT, and he had oversight responsibility for the direction of extramural studies that were funded through the NCI, which were focused on the development of new treatments for human malignant disease. The NCI has five divisions for which the NCI Director has ultimate responsibility. While working with one NCI Director from 1981 to 1988, and with another from 1988 to 1995, and during the transition year of 1988, Bruce Chabner provided stability for the DCT while many changes were occurring throughout the five divisions of the NCI. How does one assess the impact of a career on a discipline such as cancer treatment? It's not easy! Each of the articles contributed to this tribute were written by a person who trained directly with Bruce Chabner, or was otherwise directly impacted by Bruce's guidance. As can be seen from the list of contributors to these Proceedings, each individual has made major contributions to the area of cancer treatment in his or her own right. However, Bruce's contribution to cancer treatment goes far beyond the individuals he trained. The many thousands of human lives who have benefited from his efforts cannot be accurately estimated, because his contributions have been so wide-ranging, as indicated below. Being "Scientific Director" is similar in a number of ways to being a football quarterback. One of those ways is that when things go well the quarterback may get a little too much credit, and when things go not-so-well the quarterback may get too much blame. However, it is the quarterback who "calls the plays." With that in mind, a partial list of the accomplishments of the Intramural Program of the DCT while Bruce Chabner was "quarterback" includes the following: * The first human retroviruses, HTLV-1 and HTLV-2, were discovered and shown to be directly linked to the development of specific human malignancies. * Adoptive immunotherapy for human cancer was developed, offering exciting new additions to the anticancer armamentarium. * Paclitaxel (Taxol®) was developed, and shown to be the most important new anticancer agent in the past two decades. * The human genes responsible for the development of several specific malignancies were discovered, such as those for kidney cancer. * Development of blood tests to detect HIV-tainted blood. * Treatment strategies for pediatric AIDS were developed. * The AIDS Drug Development Program within the NIH was established. * New drugs for the treatment of AIDS and AIDS-related conditions were developed. * The only three drugs to date that have been specifically approved for the treatment of AIDS-AZT, DDI, and DDC-were developed under the guidance of the DCT, with Bruce Chabner as Scientific Director. * The first clinical trials conducted with each of these agents-AZT, DDI, and DDC-were performed in the Intramural Program of the DCT. * Concurrently, many of the exciting findings reported by the National Surgical Adjuvant Breast and Bowel Project over the past 10 years (as well as other cooperative groups) were a direct result of the strong support shown by Bruce Chabner during his tenure as Director of the Division of Cancer Treatment. Further, the list above does not include his personal labortory and clinical accomplishments, some of which are: * Development of the principles of use of important antimetabolites, such as methotrexate. * Elucidation of biochemical pathways affected, and the mechanisms of action, of antifols and other antimetabolites. * The conduct of seminal studies in the clinical staging of non-Hodgkin's lymphomas, using laparoscopy as a primary tool. * Important contributions to the development of multiagent regimens in the clinical treatment of lymphomas, and of Hodgkin's disease. * Developed and is editor of the textbook which is considered to be the primary reference source for anticancer chemotherapeutic agents [1]. With all of these accomplishments, his career is long from over. Having just become the Medical Director of the Cancer Center at the Massachusetts General Hospital, Bruce Chabner is uniquely poised to have an even more far-reaching impact on a discipline in which he has played such a strong seminal role. This author was never a postdoctoral fellow in Bruce Chabner's laboratory. However, more than any other single person, he has played a central role in my professional development. I know of many others for whom the same statement would be true. It is a pleasure for me to witness the launching of the second phase of an already tremendous career. From Advances in Cancer Treatment: The Chabner Symposium. Stem Cells 1996;14:64-65. PMID- 10388003 TI - Regarding: Valero V, Buzdar AU, Hortobagyi GN. "Locally Advanced Breast Cancer," The Oncologist 1996;1:8-17. AB - To the Editors: I read with interest Valero et al.'s review showing a lack of any increased cure rate over surgery from use of neoadjuvant combination therapy for locally advanced breast cancer, although a definite gain from breast conservation. Missing was any mention of hormone therapy [1] despite some of the best neoadjuvant results being from the use of Tamoxifen® in the elderly [2]. Interestingly, the highest complete remission rate in Valero et al.'s review (52%) was in a regimen including Tamoxifen® [3]. In addition, Bartlett et al. have reported that hormone therapy is superior to chemotherapy as adjuvant in estrogen receptor-positive tumors [4]. With new data from the study of radiotherapy demonstrating prostate tumor cell kill to be far higher when radiation treatment follows hormone therapy rather than when given concurrently or after therapy [5], there is a need for studies to investigate whether the same applies to hormone therapy when combined with chemotherapy in breast cancer. Further support for this comes from the latest overview of the effect of menstrual cycle timing and breast cancer surgery. The greatest gain from operating on premenopausal women occurs when the operation takes place during the progestergenic phase of the menstrual cycle, particularly in advanced node positive patients and those with high levels of progesterone pre-op attributed to high survival [6]. With increasing recognition that trauma-induced cytokine release can be a factor in tumor acceleration [7], there is a case to consider hormones followed by chemotherapy plus radiation before any major surgical interference [1], most particularly in the rare subgroup of poor-risk tumors arising during pregnancy [8]. AUTHORS' RESPONSE: We appreciate the comments of Professor Oliver regarding our review. It has been recognized for decades that breast cancer is a systemic disease. Regional and distant micro- metastases occur early, and are part of the natural history of this illness. Adjuvant chemotherapy is an integral part of the curative management of patients with primary breast cancer, including locally advanced breast cancer (LABC). The benefit derived from this therapeutic approach is well documented, and persists beyond 10 years of follow-up [1, 2]. While we recognize the paucity of studies in direct support of this statement in patients with operable LABC, a few randomized trials have been completed and published [3, 4], and the world overview of randomized trials also demonstrated this benefit [1, 2]. Several randomized studies are under way to assess the relative efficacy of primary (or neoadjuvant) versus adjuvant chemotherapy in patients with early breast cancer, as well as operable LABC. For patients with inoperable LABC the window of opportunity to perform randomized trials with local therapy only as a control arm was lost many years ago. However, the results of phase II trials compare favorably with outcomes of historical control series. We agree that adjuvant hormonal therapy is also an integral part of optimal management of selected patients with LABC (those >50 years old, and with estrogen receptor-positive tumors). The world overview demonstrated an additive effect in women treated with both systemic modalities. Therefore, for this high-risk group of patients, combined hormone and chemotherapy for those who present with hormone-responsive breast cancer might be the treatment of choice. On the other hand, the role of neoadjuvant hormonal therapy alone remains to be defined. When we last reviewed this issue [5], only limited data were available, and the results did not appear superior to local treatments without systemic therapy. We (and others) are prospectively assessing the role of neoadjuvant tamoxifen in selected patients with LABC (those who are elderly and those patients unfit for chemotherapy). Professor Oliver's comments about other important areas in breast cancer research were beyond the scope of our review. PMID- 10388004 TI - Physician Education: Myelodysplastic Syndrome. AB - CHARACTERISTICS AND PATHOLOGY OF MYELODYSPLASTIC SYNROME: Myelodysplastic syndrome (MDS) is a disease of the blood whose etiology is unclear. There is little that can be done therapeutically, and the prognosis for patients with this disease is poor. The main hematologic finding is anemia, but MDS responds poorly to the various kinds of drugs used to treat anemia, and in the past it was called refractory anemia. Moreover, 25% to 40% of MDS patients develop acute leukemia, so MDS has also been referred to as preleukemia or a preleukemic condition. When blood diseases are classified as either erythrocytic or leukocytic, it is often unclear into which category MDS falls. Although MDS sometimes occurs in young adults and children, it most often appears in older patients. Diagnosis is confirmed in laboratory tests by a reduction in peripheral blood cells, an abundance of cells in the bone marrow (cellular marrow), abnormal cellular morphology, and chromosomal abnormalities. In primary cases there is no history of underlying disease or administration of drugs that is toxic to the marrow. The course of the disease is chronic but irreversible, and in a high percentage of cases it either develops into acute leukemia or the patient succumbs to infection or hemorrhage (death due to bone marrow failure). In general, all blood cells arise from a single type of pluripotent hematopoietic stem cell in the marrow. In MDS, the hematopoietic stem cells acquire mutations and cannot produce sufficient numbers of mature blood cells (Fig. 1). In aplastic anemia the hematopoietic stem cells are also abnormal, and blood cell production in the marrow generally declines. In MDS, however, there are sufficient numbers of blood cells of each lineage along the path from hematopoietic stem cell to mature blood cell, but the cells do not completely mature and differentiate. Because of this deficiency in the differentiation process, the cells die in the marrow without maturing and differentiating (ineffective hematopoiesis). Furthermore, blood cells that escape death in the bone marrow and are released into the peripheral blood have both morphological and functional abnormalities compared with normal blood cells. In other words, MDS is an abnormality at the hematopoietic stem cell level, and it is characterized by the presence of clonal blood cells that are abnormal both in quality (morphology, function, differentiation) and quantity (cytopenia) [1]. Because these abnormalities are found in multiple blood cell lineages, they are believed to be clonal abnormalities that originate in the pluripotent hematopoietic stem cells. The reason why the stem cells become abnormal is still unclear. However, MDS can arise following treatment with antineoplastic agents such as alkylating agents or radiation treatments (therapy-related MDS), and it has been proposed that MDS is caused by cumulative DNA damage in stem cells from mutagenic substances such as antitumor drugs [2]. DIAGNOSIS AND DIFFERENTIAL DIAGNOSIS: Screening for MDS should begin with the fact that there is chronic, progressive cytopenia, the marrow is normal or hyperplastic, and there is no underlying disease (such as disseminated intravascular coagulation [DIC], portal hypertension, collagen disorder, etc.) that could otherwise cause these conditions (Table 1). MDS is most likely to occur in middle-aged and elderly patients, but because it can also occur in the young, age is not a determining factor for diagnosis. Diagnosis is verified by ineffective hematopoiesis and blood cells with morphological abnormalities in the marrow and peripheral blood. Although there are, for example, ferrokinetic studies for erythroid cells, etc., it is impossible to make an accurate evaluation of ineffective hematopoiesis based only on abnormal laboratory test results. Therefore, if chronic cytopenia and cellular marrow are both present, then abnormal morphology of blood cells becomes the deciding factor in diagnosis. Typical morphological abnormalities in MDS include megaloblasts (photo 1), dissociated maturation of the nucleus and cytoplasm (photo 2), abnormal multinucleated erythroblasts with three or more nuclei (photo 1), and ringed sideroblasts (photo 3) in the erythrocytic lineage; hypersegmented (photo 4) or hyposegmented neutrophils (pseudo Pelger-Huet nuclear anomaly, photo 5), reduced or missing granules (photos 4 and 5), and peroxidase negative neutrophils in the granulocytic lineage; and micromegakaryocytes (photo 6), megakaryocytes with multiple, isolated disc-shaped nuclei (photo 7) and giant platelets (photo 8) in the megakaryocytic lineage. However, these morphological abnormalities are not specific to MDS, and they are also seen in pernicious anemia, acute myelocytic leukemia, etc. Therefore, a diagnosis of MDS must exclude these other diseases with which we are already familiar. Once MDS is confirmed, then the type of MDS is determined in accordance with FAB classification [1] (Fig. 2). Differential diagnosis applies to all cases presenting with cytopenia. Various types of anemia such as aplastic anemia, hemolytic anemia, secondary anemia, etc., blood disorders such as idiopathic thrombocytopenic purpura, chronic neutropenia, etc., as well as collagen diseases, portal hypertension, DIC, etc., can all be differentiated from MDS based on their characteristic symptoms and laboratory test results. However, atypical forms of MDS [3, 4] also occur, such as hypoplastic marrow MDS, MDS with minimal dysplasia, amegakaryocytic MDS, etc. Meticulous microscopic examination of blood cell morphology and careful observation of the clinical course are essential, in addition to bone marrow biopsy, chromosomal studies of marrow cells, blood cell clonality analysis, etc. RECENT DEVELOPMENTS: APOPTOSIS: One biological characteristic of MDS is the presence of blood cells of abnormal clones derived from abnormal hematopoietic stem cells. These abnormal clones demonstrate ineffective hematopoiesis, which is reflected in the contradictory phenomena of normal or hyperplastic bone marrow concurrent with cytopenia in the peripheral blood. This is a result of premature cell death in the bone marrow that accompanies the abnormal blood cell differentiation found in MDS. Therefore, it has been proposed that it is very likely this early cell death takes the form of apoptosis, and, little by little, experimental results supporting this view have been published [5-7]. The development of MDS into acute leukemia is thought to be due to the survival of immature cells (blast cells) that have escaped apoptosis and have acquired the ability to proliferate [6]. Antileukemic drugs act by inducing apoptosis in leukocytes, and it is likely that acute leukemia from MDS is intractable because it has managed to bypass the mechanism of apoptosis. Research is now focused on the detection of excessive apoptosis in vivo in MDS patients and the relationship between apoptosis and the development of MDS. ORIGIN OF BLOOD CELL CLONING: Chromosomal analysis of bone marrow cells is effective as an everyday laboratory test to verify clonality, but it lacks sensitivity. The FISH method is useful for determining clonality at the level of individual blood cells, but cannot be used in patients with no chromosomal abnormalities. DNA polymorphism of enzymes mapped on the X chromosome can only be used in females, but interesting research is being conducted on the clonality of lymphocytes and the possible survival of normal hematopoietic clones. ONSET AND PROGRESSION: Unstable clones with functional deficiencies are produced by abnormal stem cells. From the standpoint of chromosomal research, it is believed that MDS occurs not from a single type of stem cell damage, but from an accumulation of multiple and random stem cell damage. MDS is a prime candidate for research on the onset of human leukemia, and when we combine what we know about MDS with its development into leukemia, we can understand the development of MDS from the standpoint of apoptosis, genetic abnormalities, chromosomal abnormalities and progression of cloning. RISK FACTORS: Many risk factors for MDS have been proposed, and it has been confirmed internationally that the four major risk factors are the blast cell ratio in the bone marrow, advanced age, chromosomal abnormalities, and thrombocytopenia. PMID- 10388005 TI - Beware the Medical-Industrial Complex. AB - ". we must guard against the acquisition of unwarranted influence, whether sought or unsought, by the military industrial complex." Dwight D. Eisenhower, 34th President of the United States (1953-1961). Farewell Address, January 17, 1961. If Ike were with us today, he might well expand his views on power and influence to include modern American medicine. The corporatization of health care in the United States has moved rapidly in recent years. Physicians are now in a position that requires us to adapt to an increasingly Darwinian existence. Years of training to be "rugged individualists" pushing the frontiers of medical knowledge have not equipped us to fight corporate battles, nor to justify our treatment decisions to bean counters. When the most important consideration becomes the bottom line, then innovation, creativity, and research diminish in importance. They will, in fact, be selected against because they cost money. Up to now, these have been the hallmarks of American medicine, and we must strive to maintain our position of American leadership in biotechnology. New developments in cancer treatment include expensive technological "bells and whistles" which physicians must ultimately evaluate objectively, despite lush advertisements from companies with obvious vested interests, and authoritative testimonials from biased investigators who presumably believe in their own work to the point of straining credulity and denying common sense. The 3-D image that was created by a computer may look beautiful (and cost accordingly), but it is hard to believe that it can fundamentally change the outcome of patients when it does not add any new data that bear on basic issues. For example, where is the exact edge of the tumor? If one pays through the nose for increasing precision where there is no new accuracy, the purchase appears less attractive, perhaps, than the hype of the salesman or the enthusiasm of a neurosurgeon or a "stereotactic" radiation oncologist (showing biased data, if any at all). For radiation therapy, the 20th century has largely represented progress by creating larger, higher energy machines for treatment. Now, with the 21st century on the horizon, x-ray treatment parameters have probably been optimized over the past 10 years or so. We see no obvious advantage in an x-ray beam beyond about 18 MeV, and none for electrons beyond 20-25 MeV. Exotic particles such as protons, neutrons, and negative pions, though expensive and difficult to deliver, have not yielded yet significant gains in either local control or survival. A variety of new afterloading machines, such as pulsed high dose rate machines, have also been developed with no clear biologic advantage over more standard remote afterloaders. Thus, new equipment will be exploiting issues of convenience, efficiency, and increased throughput (translate: economic improvement, not biological superiority). Today's technology is vastly ahead of our biologic understanding of malignant cells. Our true challenge for the 21st century is to understand the biology of malignant cells and to bring our technology to bear on the biological aspects of cancer. To improve results, cellular manipulations of some sort will probably be necessary. Perhaps these will be mediated through gene therapy, although the manipulation of some genes, to the exclusion of all others, in only tumor cells and in all tumor cells may be a biological challenge beyond our limitations. One is reminded of another time, a decade or two ago, when some tumor immunologists were predicting monoclonal antibodies would soon replace other modalities. Eventually, over time, the immunologists began to appreciate the enormous adaptability that cancer cells possess; the cells are much more than passive receptacles of antigens simply waiting to be destroyed by antibodies. Drugs which affect the function of specific oncogenes, such as the farnesyl transferase inhibitor effect on ras genes, are also quite promising. Clearly, however, there are not "magic bullets" for most cancers. The effects of gene manipulation on patient outcome, if any, are likely to be found only in the setting of combined modality therapy. The most promising clinical research from the last decade or so reinforces the utility of a combined approach in treating cancers. Unfortunately, combined treatments and the development of new combined treatments are expensive. In today's world of corporate medicine and managed care, academic centers are under considerable pressures. They are perceived as being too expensive, and thus they are in danger of being shut out of contractual arrangements with third-party representatives. If these centers are to survive, they must reform themselves: One, they must establish meaningful relationships with community hospitals and community physicians. Two, the academic programs must learn to minimize charges and deliver a true multidisciplinary service to patients in an efficient way. Three, the centers must learn to invest wisely in new technologies that community hospitals cannot and will not be expected to support. This "wisdom" refers to selection of technology that truly may have impact on the outcome of patients' lives by early detection or by treatment. The euphoria associated with projected gains of some investigational treatments can be misleading: randomized prospective trials have shown in the past that postoperative radiation following a complete resection of lung cancer, breast cancer, or rectal cancer adds a major improvement to local control, but with relatively little improvement in survival. How many times will it be necessary for companies and self-impressed investigators to rediscover this particular wheel? We must remember that every new therapy costs money, so we must focus our research time and money in promising areas. If cost is allowed to be the most important mitigator of health care, research as we know it will end. The relative lack of new therapies means that some people will die prematurely because of our lack of foresight. As scientists, we must be seen as providers of a value-added product. Improvement in cancer cure rates has been frustratingly slow. We work against a clever, tenacious adversary - both in the clinic and in the corporate board room. It is our responsibility to tout our accomplishments, admit our failures, and provide progressively better basic and clinical research with an eye toward future improvements in outcome. We must not be seen as yet another special interest come to drink at the well of public spending, but as advocates for the public good. If we fail to become important to those who control medical spending, we will be unable to make any important long-term contribution to those who matter most - our patients. PMID- 10388006 TI - Surgical Sphincter Preservation in Rectal Cancer. AB - Improved understanding of the biological features and advances in diagnostic and surgical procedures have been the basis for increased applications of sphincter preserving operations in lower rectal tumors. The relevant treatment strategies along with their indications will be presented and analyzed. New operative procedures comprise transanal excision of early rectal cancer or trans sphincteric resection or ultralow anterior resection with colon-pouch creation to improve continence. Recently, for cases in which the removal of the rectal sphincter is indispensable for oncological reasons, a continent perineal colostomy has been developed. Reconstruction of the sphinter function is achieved using a seromuscular cuff. This procedure avoids an abdominal colostomy. The neosphincter can also be formed secondarily, after a prior abdomino-perineal excison with transabdominal colostomy. PMID- 10388007 TI - Dose Intensity of Chemotherapy for Childhood Cancers. AB - Since the formulation of the "dose-intensity" concept of anticancer therapy in the mid-1980s, the concept that "more is better" has gained general acceptance among pediatric oncologists. However, recently published clinical trials results for adults with breast cancer, germ-cell tumors, and ovarian cancer raise questions about the value of dose intensification in improving outcome. Given the differences in sensitivity between pediatric and adult tumors to cytotoxic agents, the results from these adult trials suggest a need for caution but do not suggest that evaluations of dose intensity for pediatric tumors are unwarranted (especially for a tumor such as Ewing's sarcoma, which is especially sensitive to alkylating agents). Since dose-intensive therapies have significant short- and long-term costs for the patient, it is important to obtain reliable data concerning possible benefits of this strategy. Toward this end, NCI-sponsored randomized clinical trials evaluating the role of dose intensification have been initiated for several tumors of children (including neuroblastoma, germ-cell tumors, Ewing's sarcoma, and brain tumors in infants). These trials should be completed and reported in the next two to three years, and they may make unique contributions in defining the benefits and limitations of dose intensity as a cancer treatment strategy. In the long term, however, the utility of dose intensification is limited for children with cancer because of the inherent toxicities associated with its application. Identification of agents that more specifically target tumor cells is essential. Fortunately, pediatric tumor cells do have unique biological characteristics that may be susceptible to targeting for therapeutic benefit. PMID- 10388008 TI - All-Trans-Retinoic Acid Pharmacology and Its Impact on the Treatment of Acute Promyelocytic Leukemia. AB - The approach to the treatment of acute promyelocytic leukemia (APL) has changed dramatically over the past decade and, as a result, the long-term event-free survival for patients has improved significantly. The addition of the vitamin A derivative, all-trans-retinoic acid (ATRA), to treatment regimens has been responsible for this improvement in survival. Although ATRA is a potent remission induction agent in APL, continuous administration of ATRA as a single agent does not maintain patients in remission. Although lower plasma concentrations were initially noted at the time of relapse in patients with APL, subsequent studies have demonstrated that the decline in plasma drug concentrations occurs within one to two weeks of initiation of treatment, and possibly as early as three days. The inability to maintain adequate plasma concentrations of ATRA because of rapid upregulation of its catabolism is an attractive hypothesis to explain the inevitable recurrences in patients with initially responsive disease, but more recent data suggest that this mechanism alone is unlikely to be responsible for drug resistance. Cellular retinoic acid binding proteins (CRABPs) play a critical role in regulating the amount of free retinoic acid capable of reaching and activating nuclear receptors. Recent studies using leukemic blasts obtained at the time of relapse have demonstrated a shift in the ATRA dose-response curve in vitro. In addition, there is an upregulation in the expression of CRABP in leukemic blasts obtained at relapse. These observations suggest that ATRA resistance is not simply an inability to maintain therapeutic plasma concentrations of drug, but rather may be linked to the intracellular regulation of drug. The intricate nature of the homeostatic mechanisms that maintain tight control over retinoids, combined with the multiplicity of retinoid receptors and signaling pathways, leave open the possibility of a yet-to-be-defined mechanism of resistance that is independent of the clinical pharmacology of ATRA. PMID- 10388009 TI - Multiple Myeloma: An Overview in 1996. AB - Multiple myeloma (MM) has an incidence of approximately four per 100,000 per year. Ninety-nine percent of patients with MM have a monoclonal (M-) protein in the serum or urine during the course of their disease. MM must be differentiated from smoldering multiple myeloma (SMM), which has an M-protein value of more than 30 g/l and more than 10% plasma cells in the bone marrow, but no other features of MM. The plasma cell labeling index (PCLI) and the presence of circulating plasma cells in the peripheral blood help to differentiate monoclonal gammopathy of undetermined significance and SMM from MM. The current median duration of survival with chemotherapy is about three years. Patients with low PCLI and low &bgr; subset2-microglobulin values have a median duration of survival of approximately six years. Melphalan and prednisone produce an objective response in 50% to 60% of patients. Combinations of chemotherapy produce a higher response rate, but the survival rate is not different. Allogeneic bone marrow transplantation is associated with a mortality rate of 25% within six months and an actuarial survival rate of 28% at seven years. Autologous peripheral stem cell transplantation is applicable to more patients and is reported to produce a higher response rate and longer survival than chemotherapy, but most patients will eventually have relapse. PMID- 10388010 TI - In Response To: Professor Cassileth's manuscript on "Alternative and Complementary Cancer Treatments," Featured in The Oncologist 1996;1:173-179. AB - COMMENTS FOR PROFESSOR BARRIE R. CASSILETH: I have enjoyed reading the two issues of The Oncologist I've received so far. I would like to make some comments on Dr. Cassileth's article. I'm a pediatric oncologist at the National University Hospital in Singapore. Singapore is an interesting place to study people, as ours is a multi-racial country, with 75% Chinese, 16% Malays and 7% Indians, plus a significant expatriate population (Americans, British, Australians, etc.). I've been very interested in the influence of different ethnic and social backgrounds on how our patients cope with their diseases, especially families of children with cancer. We did a survey of 20 patients and found that nine of the children are given bird's nest, nine are given ginseng, and five had been given Chinese medicinal herbs. I've been doing a bit of literature search, and found that there's some evidence that Chinese medicine may help to "boost the immune system," enhancing the ability of the patients to undergo conventional cancer therapy. However, most traditional Chinese medicine practitioners would recommend that these patients continue with their "Western treatment." I have no objection to patients availing themselves of these complementary approaches, though we do not encourage it either. We are in the unenviable situation of trying to help and give advice to our patients about things about which we know little, as there is so little hard evidence. Of all the literature I've gone through so far, I find the recent Choices in Healing by Michael Lerner (MIT Press) the most helpful. PROFESSOR CASSILETH'S RESPONSE: Dr. Quah raises a number of important issues. I share his interest in the influence of culture and ethnicity on how patients and families cope with cancer. The cultural meaning of malignant disease, in fact, has long shaped not only individual reactions, but also how societies have approached research, treatment, and communication. In the United States, for example, the word "cancer" literally was banned from public print until the mid 1920s [1], and only now are physicians beginning to discuss cancer with their patients in Russia [2]. The survey of 20 patients conducted by Dr. Quah represents a substantial portion of the literature on the subject of alternative therapy use for children with cancer. During the decade of laetrile popularity in the United States, a study at a major pediatric oncology center found that 17 of 106 patients (16%) received alternative therapy, although eight different types of alternative therapies were known by 50% of parents. That study was conducted in 1977-1978 [3]. An Australian study published in 1994 [4] found that approximately 46% of children had been given alternative treatments; like Dr. Quah's patients and those in the Australian study, they remained simultaneously under mainstream care. One suspects that by now, close to 100% of parents as well as adult patients could name at least eight different types of alternative cancer therapies. I concur with Dr. Quah's assessment of Choices in Healing. It differs strikingly from other publications about alternative cancer medicine, almost all of which are proponent books that extoll unproven or discredited methods and decry a government/pharmaceutical industry conspiracy to withhold cancer cures (read "alternative, unproven methods") from the public. A visit to the health and medicine section of any large bookstore provides an eye-opening display of books on dozens of methods promoted as cancer cures. Can Chinese medicine "boost the immune system?" Claims for the mechanisms by which alternatives of the past were said to work typically reflected mainstream science of the day. The most common claim across the variety of today's popular alternative cancer therapies is that they enhance immune function. Many herbal remedies, including those from China, are sold for this purpose in America. Several Chinese medicines, such as polysaccharide from a root used in traditional Chinese medicine, Six Flavor Tea and Golden Book Tea used in conjunction with chemotherapy and radiation therapy, and Mylabris - dried Chinese beetle - have been studied for their utility against cancer in recent years. Because studies such as these are reported almost exclusively in Chinese journals, and because they tend to be preliminary, they are not well known in other countries. Dr. Quah's own professional home, the National University of Singapore (NUS), is one of the first medical schools in Asia to use the World Wide Web to distribute health information. NUS has the major cancer databases from the United States and many other important international databases. It intends to become a global health information hub on the Internet. Hopefully this resource will help fill in some of the information gaps. But most oncologists, like Dr. Quah, indeed are faced with trying to advise patients about therapies for which there is little hard evidence. The best guiding principles at this point are to discourage remedies that promise cancer cure or are promoted for use instead of mainstream treatment, encourage non invasive, comforting, complementary (adjunctive) therapies such as massage, green tea and qi gong, and check medical journals and newspapers for warnings such as those issued recently for Ma Huang (ephedrine), a still-common ingredient in herbal remedies widely available through catalogs and in health food stores. PMID- 10388011 TI - Prophylactic Oophorectomy: Reducing the U.S. Death Rate from Epithelial Ovarian Cancer. A Continuing Debate. AB - If instead of the title "Prophylactic Oophorectomy: Reducing the U.S. Death Rate from Epithelial Ovarian Cancer," the title were "Drug X Reducing the U.S. Death Rate from Epithelial Ovarian Cancer," there would be great media and medical attention worldwide to such a report. Correctly so. Regrettably, there probably is no new Drug X in the foreseeable future that will significantly reduce the death rate from ovarian cancer, be it Taxol®, taxotere, topotecan, gemcitabine, or liposomal doxorubicin-although each may result in significant responses and some prolongation of median survival. Epithelial ovarian cancer is a much more complex disease than anyone envisioned, when it was believed that extensive debulking surgery and the newest cytotoxic chemotherapy would radically reduce the death rate from ovarian cancer in the United States. Over 20 years after the first patient was treated with cisplatin for epithelial ovarian cancer, the annual death rate from ovarian cancer continued to increase. Just in the past decade, the number of women in the United States dying from ovarian cancer has increased 18% (Fig. 1) [1]. Although ovarian cancer is estimated to account for 26,700 cases and 14,800 deaths in 1996, it is a low-prevalence disease in comparison with breast cancer, which in 1996 is estimated to account for 185,700 cases and 44,560 deaths. Inexplicably, similar to breast cancer, the lifetime risk for ovarian cancer in the United States continues to increase. The most recent Surveillance, Epidemiology and End Results (SEER) calculations of lifetime risk for ovarian cancer are that 1 in 55 women will develop ovarian cancer over their lifetime, or 1.8%, up from the 1970 figures of 1 in 70, or 1.4% [2]. The 1.8% baseline lifetime risk for the general population is used to estimate the lifetime risk of known ovarian cancer risk factors (Table 1). Even utilizing what are now believed to be two of the most effective cytotoxic drugs against stage III and IV epithelial ovarian cancer, Taxol® and cisplatin, researchers reported that this resulted in an increase in the median disease-free survival of only five months, as compared with those women allocated to receive cisplatin and cyclophosphamide (median disease-free survival of 18 and 12.9 months, respectively), felt then to be the standard therapy [3]. Patients treated with Taxol® and cisplatin survived a median of 14 months longer than those treated with cisplatin and cyclophosphamide. These results may improve in women whose cancers were optimally debulked to 3.7 million donors worldwide, some patients cannot be transplanted because of the lack of a suitable HLA-identical donor. New approaches have been investigated, including the use of HLA partially mismatched, T cell-depleted, mobilized peripheral blood HSC or umbilical cord HSC. PMID- 10388068 TI - The Cure for Cancer: Not If but When. AB - As a physician specializing in the treatment of cancer, I have had to watch hundreds of patients die from cancer because current treatments have limited benefit. I know of no disease that can strike a patient more tragically than cancer. However, I can say with absolute certainty that the medical and scientific communities are on the verge of major breakthroughs in our ability to control this dreadful disease. Unfortunately, I can also say with absolute certainty that tens of thousands of Americans will die because critical research remains unfunded or underfunded, slowing the development of new lifesaving treatments by years. Never in the history of mankind have we had such an opportunity in medicine. We are literally witnessing an explosion in our understanding of cancer. Every week, new genes which regulate the process of cancer are being discovered. We now have an incredible knowledge of cancer-what makes a cancer cell a cancer cell, what cancer cells need to thrive, and what signals cancer cells to self-destruct and die. With more research our knowledge will be even greater. We also now have the remarkable ability to manipulate genes within cells, to actually direct cells to do what we want them to do. This process, one of the many new treatments we now have, is called gene therapy. This combination of increased knowledge and powerful new techniques to manipulate cells provides opportunities we could only dream of just five years ago. Let me give you just one example of how we are now using this new knowledge. I specialize in the treatment of malignant brain tumors. The most common brain tumor, the malignant glioma, is responsible for 15,000 deaths in the United States each year. The median survival time from diagnosis to death without treatment is 12 to 16 weeks. With conventional cancer treatments, including surgery, radiation and chemotherapy, the median survival is 28 weeks. Because of new research findings, we now know that malignant brain tumors are able to grow in the brain and escape destruction by our immune system because they release a protein into the brain which suppresses or "turns off" the immune system. This protein is called transforming growth factor beta (TGF&bgr;). We are able to take tumor cells and genetically engineer them in our lab so that they can no longer make TGF&bgr;, thereby uncloaking these tumor cells to the immune system. We've shown in lab experiments that rats with untreated brain tumors all died. However, rats with brain tumors treated with the genetically modified vaccine all survived. We found that rats given the vaccine were able to develop immunity against these tumors and their brain cancer was completely eradicated. Based on these studies, we now have a clinical trial where tumor cells are removed from patients during surgery, genetically engineered to make a cancer vaccine, and then re-injected into patients with brain cancer. Six weeks ago we treated the first patient with the vaccine, a 36-year-old man with three young children whose brain tumor was growing despite two brain surgeries and radiation therapy. His tumor appeared to have stabilized after his first vaccine injection. This is just one of literally hundreds of novel approaches now under development for the treatment of cancer. When plans to start this experimental trial were first announced a year ago, my office received over two thousand phone calls, faxes, and e-mails from desperate patients hoping to participate in this trial because they had failed conventional treatments. Due partially to limited funding the trial started just six weeks ago. I would venture that most of the patients who called my office have now died. Even with the study under way, only twelve patients can be entered into the trial, out of potentially thousands who could be treated. This is the most painful reality, knowing that our patients will die because our country has not made cancer research a higher priority. To continue rapid progress requires increased funding for not only basic research to continue our understanding of cancer, but also for the translation of research into clinical trials for patient care. Our national budget for cancer research should be at least twice the current funding levels. We no longer wonder if we will find a cure for cancer but when. America has an incredible opportunity to conquer this deadly disease. Increasing funds for cancer research could now accelerate by years the development of new and more effective treatments for cancer, literally saving tens of thousands of American lives. PMID- 10388069 TI - Adjuvant Drug Treatment for Resectable Breast Cancer. AB - Breast cancer is the most common life-threatening malignancy in Western women and the second most common cause of cancer-related death. A paradox in the care of patients with breast cancer is the observation that the majority appear to be curable at the time of initial surgery, yet a large number later experience relapse followed by death from disease. To combat this problem, systemic drug therapy in conjunction with surgery and radiation therapy is now standard for many patient subgroups. Standard medical treatment options include up to five years of tamoxifen for receptor positive amenorrheic patients, approximately six months of combination chemotherapy for younger patients and those who are receptor negative, and both in some patient subgroups. These interventions have a profound public health impact even though the majority of patients destined to recur do so even following "optimal" treatment. Further improvements are hoped for as a result of ongoing research involving the use of high-dose chemotherapy, dose-dense chemotherapy, and newer agents. Clinical trials testing these approaches offer the best chance to continue our progress and help to better define appropriate and standard treatment strategies for the future. PMID- 10388070 TI - Topoisomerase I Inhibitors. AB - Topoisomerase I inhibitors are a new class of anticancer agents with a mechanism of action aimed at interrupting DNA replication in cancer cells, the result of which is cell death. Most if not all Topoisomerase I inhibitors are derivatives of the plant extract camptothecin. Irinotecan (CPT-11), a semi-synthetic derivative of camptothecin, is approved in the United States for the treatment of colorectal cancer. Ongoing clinical trials with CPT-11 show a 13% to 32% response rate when it is used singly or in combination with other chemotherapeutic agents such as 5-fluorouracil. The major dose-limiting toxicities of CPT-11 are myelosuppression and a dual phase diarrhea. Topotecan is another semi-synthetic analogue of camptothecin. It is approved for use in the United States for the treatment of cisplatin refractory ovarian carcinoma. Current clinical trials suggest antitumor activity against a variety of human tumor types. There is significant interindividual variability in the plasma disposition of this drug. The main dose-limiting toxicity is myelosuppression. There are other derivatives of camptothecin, as well as new formulations of the parent plant extract, that are in various stages of clinical trials. Some of these clinical trials are aimed at increasing the therapeutic benefits of the agents when used singly or in combination with other chemotherapeutic agent(s) or treatment modalities. The dose-limiting toxicity observed in most of these clinical trials is myelosuppression. PMID- 10388071 TI - Outpatient Management of Febrile Neutropenia: Should We Change the Standard of Care? AB - The syndrome of fever and neutropenia is an iatrogenic complication of cytotoxic therapy for cancer. Because febrile neutropenia is associated with serious infection, patients with the syndrome are treated emergently with broad-spectrum, high-dose antibiotics. Recently, a differentiated approach to febrile neutropenia has been explored, based on assessment of risk. Prediction rules to identify low risk patients were developed, and outpatient management of low-risk patients has been explored. Based on pilot studies and early randomized trials, some have called for a new standard of outpatient care. This article describes the scientific rationale for the current standard of care for febrile neutropenia; reviews the risk assessment studies; discusses the issues of safety, quality of life, and shifting of economic burdens arising when outpatient care is substituted for inpatient care-a change in practice patterns sweeping through acute care medicine with little scrutiny; and critically reviews the published trials of outpatient treatment of febrile neutropenia. PMID- 10388072 TI - Childhood Acute Lymphoblastic Leukemia. AB - The cure rate for childhood acute lymphoblastic leukemia (ALL) now exceeds 70%. This success has been achieved in part by improvements in the biologic characterization of newly diagnosed patients. Modern treatment protocols rely on this information to tailor therapy to a patient's risk of relapse. Patients with favorable genetic features, such as hyperdiploidy or the TEL-AML1 fusion, can be treated with conventional antimetabolite-based therapy to minimize long-term side effects. By contrast, extremely high-risk patients, such as infants with MLL gene rearrangements and cases with BCR-ABL fusion and poor early response, are candidates for allogeneic hematopoietic stem cell transplantation in first remission. Future areas of research include the identification of new genetic subgroups of ALL and the development of novel therapies. PMID- 10388073 TI - Positron Emission Tomography: Current Role for Diagnosis and Therapy Monitoring in Oncology. AB - High-resolution cross-sectional imaging improved dramatically the diagnosis and therapy management of oncological patients, although several questions remained open, primarily concerning the exact initial staging, the differential diagnosis of recurrent tumors, and therapy management. Positron emission tomography (PET) is a quantitative, functional imaging modality from the field of nuclear medicine which has the potential to yield physiological information. The diagnosis of tumors with PET is based on the increased regional glucose metabolism. Furthermore, PET can serve as a valuable tool for monitoring therapeutic effects. The most common tracer used for oncological studies is F-18-deoxyglucose (FDG), a glucose analog. FDG-PET has been evaluated in different tumor types such as colorectal cancer, malignant lymphomas, melanomas, soft-tissue sarcomas, and lung tumors for both the diagnosis of primary tumors and recurrent lesions. The sensitivity of PET-FDG studies exceeds 85%, dependent on tumor type, size, and location. The diagnosis of viable tumor tissue following chemotherapy is another application of PET. A limitation of FDG-PET studies is false positive results, e.g., due to inflammation. This problem may be overcome by the use of multitracer studies and/or labeled amino acids. Different procedures can be used to evaluate therapeutic effects. FDG follow-up studies are used to assess early effects and to predict long-term response. Pharmacokinetic studies of labeled therapeutic agents such as F-18-fluorouracil or C-11-ethanol provide exclusively quantitative data about transport and elimination of a drug. PET with labeled cytostatic drugs permits a prognosis prior to onset of chemotherapy. This procedure is recommended for monochemotherapy. In patients receiving polychemotherapy, the evaluation of different resistance mechanisms is needed and new approaches using suitable substrates, e.g., for the P-glycoprotein, are being developed or are already in use for scientific purposes. PMID- 10388074 TI - Understanding the Myelodysplastic Syndromes. AB - The myelodysplastic syndrome (MDS) remains challenging to the clinician in terms of diagnosis and management. The diagnosis is essentially one of exclusion in first ruling out other disorders that can also cause peripheral blood/bone marrow cell dysplasia and cytopenias. The distinguishing biological characteristic of MDS is that it is a clonal disorder of the marrow with impaired differentiation. Recent studies implicate extensive apoptosis as the explanation of the paradoxical observation of marrow hyperplasia but peripheral blood cytopenia. Neutropenia and/or neutrophil dysfunction account for the primary clinical manifestation of MDS in terms of an increased risk for infection, which is the leading cause of death in MDS. The clonal nature of MDS places it also at continual risk for transformation to acute leukemia. Predicting overall survival as well as the risk of AML transformation has been improved by the recent development of a scoring system (International Prognostic Scoring System) that incorporates three laboratory variables: percent of marrow blasts, degree of cytopenias, and presence of chromosomal abnormalities. Based on these variables, four prognostic subgroups can be delineated ranging from low risk with a median survival of 5.7 years, to high risk with a median survival of 0.4 years. Management of MDS can now be based on the patient's respective prognostic subgrouping, with low-risk patients being considered for hematopoietic growth factor singly or in combination if at the point of red cell transfusion dependence and/or neutropenia with recurrent infections, while high-risk patients should be offered AML-induction therapy or novel agents such as topotecan. One must individualize further in patients in the remaining intermediate groups, I and II, in choosing the most appropriate therapy. Future advances upon understanding the molecular details of the MDS clone should ultimately improve the care of patients with MDS. PMID- 10388075 TI - Clinical Use of Irinotecan: Current Status and Future Considerations. AB - Irinotecan (CPT-11, Camptosar), a semisynthetic, water-soluble derivative of the plant alkaloid camptothecin, is a drug which has undergone extensive clinical investigation worldwide. It is, at this time, commercially available in the United State for the treatment of fluorouracil-refractory colorectal cancer. In this review, I will discuss the current approved clinical use, discus the issues of toxicity and its management, and consider some of the ongoing clinical investigations which are exploring possible future uses for this agent. PMID- 10388076 TI - CD34+ Selection: The Basic Component for Graft Engineering. AB - The hematopoietic system consists of a network of stem and progenitor cells of varying degrees of maturity interacting with other cells that act in supportive and regulatory capacities. Many cell types have been identified by their in vivo or ex vivo growth potential, i.e., colony assays, or by physical characteristics like cell-surface markers which can be identified by monoclonal antibodies. One of the most studied antibody markers is the CD34 antigen which is present on the most primitive hematopoietic progenitor cells. PMID- 10388077 TI - The Cost of Biological Terrorism. AB - Most Americans are or will be facing a threat more real than crimes or terrorism it is the threat of cancer. Indeed, one-fourth of all Americans alive today will ultimately die from cancer. Yet the level of funding for cancer research in 1998 and beyond remains in doubt. The Senate Appropriations Committee has proposed a higher funding figure than the House and the difference will be resolved in negotiations this September. President Clinton has recommended a meager 2.5% increase in spending on cancer. This sum is simply not enough. Although Americans may fool themselves into thinking the government has been at war against cancer, the current funding ceiling for the National Institutes of Health (NIH) budget demonstrates that this so-called offensive is little more than a skirmish. Careful scrutiny of this budget reveals that every time a citizen pays ten dollars in taxes, only one penny goes to cancer research. What the government is spending in cancer research would not buy or maintain two stealth bombers-hardly evidence of a major military strike. For those of us fighting the deadly scourge of cancer, the 1,550 Americans killed each day by this disease are painful and enduring casualties. Imagine five fully loaded jumbo jets crashing with no survivors on the same day. These headlines would generate fast and effective calls for funding for improved safety regulations, and yet cancer takes this number of lives daily-and in one year more lives than all the U.S. combat fatalities in this century. And yet there is a virtual silence as Congress meets to determine the level of monies to direct to cancer research efforts which might halt this carnage. In the past, medical research stopped the horrors of pain associated with amputations and operations conducted without anesthesia, and research stopped the epidemic of polio and the massive deaths from typhoid fever. Soon research will stop the deaths from AIDS. Will cancer be prevented or cured within your lifetime? It should and can be, but only if Americans speak up and demand Congress do its part to advance promising research by supporting NIH and efforts to control cancer. Cancer has stricken almost every family and we urgently need more defense. We must not sit waiting quietly any longer. It is time for a wake-up call to arms, to declare a war effort that demands results. The U.S. paid thirty times more for the Gulf War than the annual budget allotment for cancer research, and five times more to maintain the space program. Today, three-fourths of all cancer research grants approved by critical scientific review still go inactive. These instances represent more lost opportunities, more lost time, more lost lives. We must do better than this. Cancer strikes at the heart and fabric of our society. For the 1.3 million Americans diagnosed with cancer this year, the cost to our economy will exceed $100 billion. Thus, the $2.4 billion now being spent on research is insufficient medically and morally. Several clear-sighted congressional leaders who understand the critical need for increased funding have worked to assure expanded resources for the NIH and for cancer research. Unfortunately, their voices are too often drowned out by legislators with different agendas. Even with limited funds, America's past 25 years of cancer research has paid off. We have already cured some types of cancers, especially those that strike our young people. From 1973 to 1990, the cancer death rate from birth to 19 years of age decreased 38%; from 20 to 40 years, 20%; and from 45 to 54 years, 10%. Over this same period, the government invested $56 million on testicular cancer research. This effort yielded a 91% cure rate and produced an annual savings of $166 million that will last forever. The financial investment was repaid in six months, and the victims have an increased life expectancy of 40 years. These examples and others are proof of the principle that the support of cancer research pays off. However, we have not yet won such hard-fought victories on the more prevalent forms of cancers such as breast, prostate, and lung. Whether it is by cure or prevention, cancers must also be controlled and it can only be accomplished through research. On December 23, 1971, President Richard Nixon signed the National Cancer Act that was to provide, ".a total commitment of Congress and the president. to provide the funds. for the conquest of cancer." Somewhere this contract with America got lost. We are now faced with more losses. The new upheaval in American medicine threatens clinical research in cancer investigation to test new therapies and to support the training of new research soldiers to join the battle. The bottom-line approach of insurance and managed care policies no longer cares about these essential components; they say it is no longer their responsibility. This loss of financial support, combined with the tepid endorsement of funding of research from the government, occurs just when there is an explosion of new discoveries and opportunities becoming available to attack cancer. It is time to dramatically increase our efforts and no time to retreat. If we really want to defend against the terrorism of cancer, we need to attack it with a real war effort. If we can go to Mars, we can go to war on cancer, but only if Americans can speak louder than their elected government leaders. They need to hear our opinion, now. PMID- 10388078 TI - Topotecan: Incorporating It Into the Treatment of Solid Tumors. AB - This issue of The Oncologist provides the reader with two useful reviews of the new chemotherapeutic agent topotecan, one of a class of topoisomerase I inhibitors that is being studied and incorporated into the treatment of various malignancies. Topotecan was approved for the treatment of refractory ovarian cancer in 1996, and has shown promising activity against a variety of solid tumors, as well as hematologic malignancies. One paper discusses clinical guidelines for managing topotecan-related hematologic toxicities, and centers on data derived from ovarian cancer studies. The other focuses on the role of topotecan in the treatment of small cell lung cancer (SCLC), where it has consistently shown encouraging results and for which definitive trials are now being conducted. As front-line therapy for ovarian cancer and small cell lung cancer is dominated by platinum-based regimens, the dosing guidelines and management issues discussed are pertinent for both tumor types. PMID- 10388079 TI - Clinical Guidelines for Managing Topotecan-Related Hematologic Toxicity. AB - PURPOSE: Topotecan, a semisynthetic water-soluble camptothecin analog, was recently approved as a second-line treatment for women with ovarian cancer. In clinical trials, hematologic toxicity has been the predominant toxicity associated with its use. The purpose of this article is to provide guidelines on the clinical management of these toxicities. METHODS: The guidelines on the management of hematologic toxicities associated with topotecan therapy for advanced ovarian cancer patients were established through a review and analysis of phase I, II, and III clinical trials. RESULTS: In phase I studies, noncumulative neutropenia was the predominant toxicity associated with topotecan therapy. In subsequently conducted phase II trials, thrombocytopenia related to prior carboplatin and alkylating agent therapies has become a prominent toxicity, and neutropenia has been more severe than anticipated from phase I studies. The risk for both toxicities relates to the extent of prior myelosuppressive chemotherapy and to renal impairment. These toxicities can be managed through the identification of high-risk patients and implementation of appropriate prophylactic measures. Such measures include dose reductions or the use of hematopoietic growth factors. For patients with persistently low blood cell parameters, transfusion therapy remains a viable option. CONCLUSION: Hematologic toxicities associated with topotecan therapy are noncumulative. Consequently, once a dosing regimen is established, toxicity patterns are predictable. Pretreatment assessment of the nature and toxicities of prior therapy and renal function should assist the clinician in preventing complications caused by the myelosuppressive effects of topotecan therapy. PMID- 10388080 TI - The Role of Topotecan in the Treatment of Small Cell Lung Cancer. AB - Topotecan is a chemotherapeutic agent that is active in the treatment of small cell lung cancer (SCLC). As a first-line agent in chemotherapy-naive patients with extensive disease SCLC, topotecan has a 39% response rate. As a second-line drug in SCLC patients with "sensitive" disease and "refractory" disease, the response rate is greater than 38% and less than 10%, respectively. The combination of topotecan and paclitaxel exhibits a promising overall response rate of 92% in chemotherapy-naive patients with extensive disease SCLC. Further studies are warranted with topotecan used in combination with other agents, including radiation therapy in patients with SCLC. PMID- 10388081 TI - New Perspectives on an Old Friend: Optimizing Carboplatin for the Treatment of Solid Tumors. AB - BACKGROUND. Since its clinical introduction in 1981, carboplatin has proved a feasible alternative to cisplatin for the treatment of many solid tumors, especially ovarian cancer. Because the pharmacokinetics and, ultimately, the pharmacodynamics of carboplatin are highly dependent on the status of renal function, fixed dosing based on body surface area has led to carboplatin overdosing or, especially, underdosing. This has resulted in less than optimal treatment results compared with cisplatin in a variety of solid tumor types. Only in the past few years has the optimal dosing method for carboplatin individualizing the dose (area under the concentration-versus-time curve [AUC]) rather than conventional use of body surface area-been adopted by clinical oncologists. METHODS. An extensive review of the oncology literature has been performed to update both carboplatin dosing guidelines as well as its present role in solid tumor chemotherapy. Initial efforts to devise a dosing formula for carboplatin focused on reducing myelotoxicity (especially thrombocytopenia). Subsequently, a simple formula was developed to adjust the carboplatin dose according to renal function. By targeting a carboplatin AUC rather than empirically dosing according to body surface area, doses of carboplatin can be individualized to fall within the drug's therapeutic index. RESULTS. The use of carboplatin dosing guidelines based on renal function has led to optimization of its pharmacodynamic effects both with respect to its safety profile and its ultimate impact on solid tumor response and patient survival. Since carboplatin has little neurotoxicity, it has become the platinum agent of choice in combination with paclitaxel for therapy of previously untreated ovarian cancer. Carboplatin plus etoposide and carboplatin plus paclitaxel have been studied in phase II and III trials, with the latter combination demonstrating improved activity against advanced non-small cell lung cancer. Additional trials in patients with other solid tumors have shown that carboplatin is more cost effective and less toxic than cisplatin. CONCLUSIONS. Dosing based on renal function and a targeted serum AUC, rather than on body surface area, has resulted in the optimal utilization of carboplatin in cancer chemotherapy. Its predictable toxicity and clinical efficacy equivalent to cisplatin make carboplatin the drug of choice for selected tumor types. PMID- 10388082 TI - Adjuvant Chemotherapy for Resected Non-Small Cell Carcinoma of the Lung: Why We Still Don't Know. AB - The role of adjuvant chemotherapy in resected non-small cell carcinoma of the lung (NSCLC) remains controversial. A critical review of the 11 cisplatin-based randomized trials addressing this issue was performed using methodology adapted from the CONSORT statement. The 11 trials were divided into those that included predominantly node-negative patients (n = 5) and those that included predominantly node-positive patients (n = 6). In the node-negative trials, which included 1,084 evaluable patients, no evidence of heterogeneity of treatment effect between the trials was found. The composite five-year survival rate for chemotherapy patients was 61% versus 55% for control patients, which reached marginal significance (p = 0.06). In the node-positive trials, which included 880 evaluable patients, a marginal lack of homogeneity of treatment effect between trials was found (p =.013). The composite two-year survival rate for chemotherapy patients was 48% versus 40% for control patients, which reached marginal significance (p = 0.06). Although not definitive, these trials suggest a benefit to postoperative adjuvant chemotherapy in resected NSCLC. The cumulative experience of the 11 trials is notable for: A) the small number of patients; B) heterogenous patient populations; C) substandard chemotherapy regimens; D) inadequate chemotherapy delivery, and E) less than optimally executed clinical trials. The 11 trials are discussed in detail, and recently completed and ongoing trials of adjuvant chemotherapy in resected NSCLC are reviewed. PMID- 10388083 TI - Core Biopsy as Alternative to Fine-Needle Aspiration Biopsy in Diagnosis of Breast Tumors. AB - In spite of the widespread use of cytological smears for diagnosis of breast cancer lesions, many surgeons are still reluctant to accept the cytological report as the only criterion for performing definitive surgery. Modern surgical strategy requires a preoperative planning of the surgical treatment, possible through the use of core biopsy, which provides a diagnosis based on tissue specimens, thus permitting the study of both the architectural and cytological patterns. The authors report their five-year experience with this technique and evaluate its diagnostic usefulness and ability to reduce intraoperative biopsy procedures. The histological examination of 92 palpable breast lesions, clinically and mammographically detected, was performed with core biopsy, and diagnosis was confirmed with the surgical sample in 80 cases. A definitive histological diagnosis was obtained with core biopsy in 90% of cases. Only nine cases required confirmation with frozen section diagnosis at the time of definitive surgery. The sensibility of core biopsy was 92%, specificity and predictive value of positive result were 100%, and diagnostic efficiency was 86%. This study confirms the usefulness of systematic use of core biopsy for definitive preoperative diagnosis of breast cancer; the simplicity, safety and low cost of this method also make ultrasound-guided core biopsy applicable to nonpalpable lesions. PMID- 10388084 TI - Octreotide Acetate in the Treatment of Fluorouracil-Induced Diarrhea. AB - Cytotoxic chemotherapy, particularly the regimens that contain 5-fluorouracil (5 FU), can produce diarrhea. Octreotide acetate appears to have a major therapeutic effect in the management of 5-FU-induced diarrhea. A prospective study was conducted to investigate the efficacy of two different doses of octreotide acetate, 100 ug and 500 ug three times daily, for the treatment of severe 5-FU induced diarrhea refractory to loperamide, and also to evaluate whether the higher dose is more effective in the management of this complication. Fifty-nine patients with tissue-documented colorectal and head and neck carcinoma were enrolled in this study, 28 in the 100 ug arm and 31 in the 500 ug arm of octreotide acetate which was administered s.c. three times daily. Patients were required to have National Cancer Institute Common Toxicity Criteria 90% of cases. The morbidity of sentinel lymphadenectomy is minimal, considerably less than the 15%-20% rate of complications associated with axillary lymph node dissection. Moreover, excision of the sentinel node provides a specimen for focused histopathologic analysis and experimental studies using sensitive immunohistochemical techniques and even reverse transcriptase polymerase chain reaction, which may improve detection of axillary metastases. Intraoperative mapping of the lymphatic tract draining to the sentinel node may use vital blue dye and/or radioactive tracer. The rate of sentinel node detection exceeds 90% with either agent alone or in combination. Because definitive follow-up data are not yet available, intraoperative lymphatic mapping and sentinel lymphadenectomy should be considered an experimental staging adjunct rather than a therapeutic modality. PMID- 10388101 TI - Systemic Chemotherapy in Gastric Cancer: Where Do We Stand Today? AB - Gastric cancer is one of the major causes of cancer-related mortality worldwide. Its prognosis is poor, and surgery offers the only realistic chance of cure. Nevertheless, most of the patients present with inoperable tumors, while the recurrence rate after potentially curable resections is high. In these patients, systemic chemotherapy has been used for palliation of symptoms and possibly for prolongation of survival. 5-fluorouracil (5-FU) is the most widely used agent in chemotherapy of gastric cancer alone or combined with other cytotoxic drugs. Until recently, combination chemotherapy produced modest results, with no significant impact on survival. Progress in research studying the mechanisms of action of various chemotherapeutic agents led to the design of more active chemotherapy regimens. Combinations of 5-FU and cisplatin and the use of modulators of 5-FU activity have produced high response rates, including complete responses in more than 10% of patients with advanced gastric cancer, and, in certain studies, a small but significant survival benefit over older regimens. Adjuvant chemotherapy has not generally produced a significant survival benefit in patients undergoing curative resection. The use of newer, more effective regimens is currently being investigated and might prove useful in certain high risk groups. Neoadjuvant chemotherapy, chemoradiotherapy, and chronomodulated administration of 5-FU, along with the use of novel chemotherapeutic agents, represent exciting areas for clinical research which might further improve the role of systemic chemotherapy in gastric carcinoma. PMID- 10388102 TI - Pancreatic Cancer: Local Success and Distant Failure. AB - The cure rate for pancreatic cancer remains less than 5% despite more than 20 years of clinical trials. Nevertheless, a select group of patients benefit from therapy at all stages of disease and important concepts regarding patient care have emerged. The development of agents such as gemcitabine and docetaxel have spurred a new generation of clinical trials in pancreatic cancer. An appreciation for the results of the many adjuvant and neoadjuvant trials and the application of lessons learned in the care of these patients is necessary to design the new trials. PMID- 10388103 TI - Prognostic Factors in Aggressive Non-Hodgkin's Lymphomas. AB - Aggressive non-Hodgkin's lymphoma (NHL) is a biologically heterogeneous disease that can be cured with aggressive chemotherapy treatment. Different clinical, biological, cellular and molecular features have been identified as having prognostic significance on the outcome of NHL patients. The knowledge of these prognostic features can be used in everyday practice in order to predict the prognosis of every new NHL patient and tailor his or her treatment accordingly. PMID- 10388104 TI - Gene Regulation and Clinical Roles for Interferons in Neoplastic Diseases. AB - OVERVIEW: Clinical indications for the use of interferons (IFNs) for cancer continue to expand and will likely continue to do so. IFNs have been approved for clinical use by the United States Food and Drug Administration for chronic myelogenous leukemia (CML), hairy cell leukemia, follicular lymphomas, Kaposi's sarcoma in the setting of AIDS, and melanoma for patients at high risk for recurrence after surgery. In addition, as a result of their antiviral activity, IFNs result in control of chronic active hepatitis and recurring papillomas that may reduce cancer development resulting from these processes and their underlying viruses. For almost all of these indications, therapeutic activity has been established from well-conducted, international phase III clinical trials. IFNs were the first successful biological therapy for human malignancy; they can synergize to produce tumor regression with surgery and chemotherapy and can potentiate other cytokines and monoclonal antibodies. IFNs and cytokines can modulate gene expression, resulting in enhanced immune effector-cell number, cytotoxicity, antigen expression, and production of other cytokines. IFNs have pleiotropic effects on cellular function, including influences on growth, differentiation, and immunologic function. For greatest effects, IFNs are used in combination with other modalities of therapy. This increases the effect of IFNs or allows IFNs to increase the effects of other therapies. Cytosine arabinoside improves the therapeutic effectiveness of IFNs in CML, and IFN-&agr;2b improves the prognosis, survival, and quality of life after surgery for high-risk patients with melanoma. Gene modulation by IFN-&agr; or IFN-&bgr; of thymidine phosphorylase, an enzyme important in DNA synthesis, has been suggested to be the basis for enhancing 5-fluorouracil (5-FU) effectiveness in preclinical models and may augment effectiveness in adenocarcinomas. IFNs increase the expression of some tumor-associated antigens that could be of benefit for combination use with monoclonal antibodies for imaging or therapy. PMID- 10388105 TI - "Just Another Statistic" AB - On returning from a medical meeting, we learned that sadly a patient, "Mr. B.," had passed away. His death was a completely unexpected surprise. He had been doing well nine months after a course of intensive radiotherapy for a locally advanced head and neck cancer; in his most recent follow-up notes, he was described as a "complete remission." Nonetheless, he apparently died peacefully in his sleep from a cardiac arrest one night and was found the next day by a concerned neighbor. In our absence, after Mr. B. expired, his death certificate was filled out by a physician who didn't know him in detail, but did know why he recently was treated in our department. The cause of death was listed as head and neck cancer. It wasn't long after his death before we began to receive those notorious "requests for additional information," letters from the statistical office of a well-known cooperative group. Mr. B., as it turns out, was on a clinical trial, and it was "vital" to know further details of the circumstances of his passing. Perhaps this very large cancer had been controlled and Mr. B. succumbed to old age (helped along by the tobacco industry). On the other hand, maybe the residual "fibrosis" in his neck was actually packed with active tumor and his left carotid artery was finally 100% pinched off, or maybe he suffered a massive pulmonary embolism from cancer-related hypercoagulability. The forms and requests were completed with a succinct "cause of death uncertain," adding, "please have the Study Chairs call to discuss this difficult case." Often clinical reports of outcomes utilize and emphasize the endpoint "disease specific survival" (DSS). Like overall survival (OS), the DSS can be calculated by actuarial methods, with patients who have incomplete follow-up "censored" at the time of last follow-up pending further information. In the DSS, however, deaths unrelated to the index cancer of interest are censored at the time of death; thus, a death from intercurrent disease is considered a "success" (to the investigator, that is; obviously, not to the patient and his or her family). The DSS rate will always be superior to the OS rate. Obviously, for any OS curve, if one waits long enough it will ultimately come to zero. There is thus a very logical rationale for reporting the DSS separately, particularly in diseases where death from intercurrent disease is expected to be common. Analyzing the DSS allows researchers to better compare the biologic efficacy of two or more cancer treatments, since it does not necessarily come to zero. Unlike some other endpoints, including local-regional control or freedom from progression, it takes into account the possibility of salvage therapy. DSS also focuses on an endpoint of interest to the public-death from cancer. In a recent popular media survey in which people were asked how they would choose to die if they could, 0% selected cancer. However, there are two serious potential problems with heavy dependence on the DSS. First, since patients who die from intercurrent disease are considered "cured," it seriously inflates the apparent effectiveness of a cancer treatment. Given the same biologic disease and the same treatment, the DSS as calculated in an old, sick population at high risk of intercurrent death will be better than the DSS in a younger, healthier population whose major risk is from their cancer. This problem has been discussed with respect to early stage prostate cancer, in which the conservative approach of observation has been criticized. The studies at issue rely heavily on the DSS, suggesting a comparable DSS (90% at 10 years) with "watchful waiting" to other researchers' results with aggressive therapy. The problem is that these series of conservative management focus on a patient population (as opposed to individuals) with a high risk of competing causes of mortality, which is very different from the population of patients generally treated with aggressive therapy (in which some have shown overall survivals superior to age-matched controls). It is fallacious and illogical to compare nonrandomized series of observation to those of aggressive therapy. In addition to the above problem, the use of DSS introduces another potential issue which we will call the bias of cause-of-death-interpretation. All statistical endpoints (e.g., response rates, local-regional control, freedom from brain metastases), except OS, are known to depend heavily on the methods used to define the endpoint and are often subject to significant interobserver variability. There is no reason to believe that this problem does not occasionally occur with respect to defining a death as due to the index cancer or to intercurrent disease, even though this issue has been poorly studied. In many oncologic situations-for example, metastatic lung cancer-this form of bias does not exist. In some situations, such as head and neck cancer, this could be an intermediate problem (Was that lethal chest tumor a second primary or a metastasis?.Would the fatal aspiration pneumonia have occurred if he still had a tongue?.And what about Mr. B. described above?). In some situations, particularly relatively "good prognosis" neoplasms, this could be a substantial problem, particularly if the adjudication of whether or not a death is cancer-related is performed solely by researchers who have an "interest" in demonstrating a good DSS. What we are most concerned about with this form of bias relates to recent series on observation, such as in early prostate cancer. It is interesting to note that although only 10% of the "observed" patients die from prostate cancer, many develop distant metastases by 10 years (approximately 40% among patients with intermediate grade tumors). Thus, it is implied that many prostate cancer metastases are usually not of themselves lethal, which is a misconception to anyone experienced in taking care of prostate cancer patients. This is inconsistent with U.S. studies of metastatic prostate cancer in which the median survival is two to three years. It is possible that many deaths attributed to intercurrent disease in "watchful waiting" series were in fact prostate cancer related, perhaps related to failure to thrive, urosepsis, or pulmonary emboli. We will not know without an independent review of the medical records of individual patients; in some cases, even the most detailed review, sometimes even an autopsy, will not be conclusive. There are only a few data available describing the problems created by cause-of-death-interpretation bias. One small study, presented only in abstract form, assessed the cause of death in 50 randomly selected prostate cancer patients who died. Five experts in prostate cancer were asked to assign the cause of death as due to or not due to prostate cancer. The DSS varied from 21% to 35% among the five reviewers, a relative difference of 66%. Studies of autopsies, which are now rarely done in the U.S., have shown that fatal malignant tumors were occasionally missed by clinicians and-even more sobering-an occasional patient thought to have died from metastatic cancer is found to have no tumor but to have died from a "benign" cause such as TB. One study suggested an error rate of approximately 8%. Clearly the use of DSS is here to stay and is a useful adjunct to OS in analyzing randomized trials. There needs to be more research on the validity and interobserver reproducibility of the DSS. In the meantime, researchers should not report DSS without reporting OS and the reasons for intercurrent deaths should be described-peer reviewers should enforce this. As with so many other problems with statistics in the medical literature, it is the job of the reader to remain skeptical. The rate of intercurrent deaths in a study should reflect the age and demographics of the study population. If the DSS is far superior to the OS, the population being studied may be unusually sick (and thus unrealistic), or there may be a bias in classifying the causes of death. Similarly, if the DSS and OS are identical (unless a highly virulent malignancy is being studied), it may suggest the researchers have only included an unusually healthy (and thus unrealistic) patient population. Finally, we would also be a bit suspicious of a sizeable series that did not have any deaths that were considered of "uncertain" cause, unless the researchers specifically included them as being due to the cancer. We honestly think that everybody has a few patients like Mr. B. PMID- 10388106 TI - Arsenal of Hope: Revolution in Genetics Creates New Weapons in the War on Cancer. AB - [The following article appeared in The Wall Street Journal, issue of May 6, 1998. Blocking a 'Helper Protein'] The war on cancer, fought for three decades marked by failure and frustration, suddenly is in overdrive. Just a few weeks ago, reports rocked the medical world about two drugs that show promise in preventing breast cancer. This week, Wall Street and Main Street alike went wild over word of a bold experimental drug that wiped out tumors in mice. These approaches are in very early stages of development and, even if all goes well, will require years of human testing before they can move into widespread usage. But they underscore a much bigger story: A quiet revolution in genetics has brought scientists closer than ever before to finding an actual cure for cancer. The drugs that made headlines this week use the promising approach of blocking a tumor's blood vessels. Even farther along in development, however, is a whole new generation of gene-based drugs aimed at a strikingly broad range of cancers. Human testing is already under way for this new arsenal, which looks to be far more powerful and far less toxic than anything tried before. PMID- 10388107 TI - Some Aim to Fix the Genes Themselves. AB - [The following article appeared in The Wall Street Journal, issue of May 6, 1998.] Ever since scientists learned over a decade ago that cancer is the result of defective genes, they have dreamed of shutting down tumor growth simply by replacing the bad genes with good ones. Researchers at two biotech companies and a pharmaceutical giant believe they are close to making that dream come true, at least for some patients. In two weeks, scientists will present results of several studies showing, for the first time, that cancer growth in severely sick patients can be stalled through an innovative method of repairing damaged genes. If the initial studies of this "cancer gene therapy" in about a hundred patients hold up, one of the companies, closely held Introgen Therapeutics Inc. of Austin, Texas, believes its technique may be available to doctors in two years. PMID- 10388108 TI - p53: The Challenge of Linking Basic Science and Patient Management. AB - Abnormalities of the p53 tumor suppressor gene are the single most common molecular abnormality seen in human cancer, being found in more than 50% of malignancies. Considerable evidence indicates that the product of this gene has critical roles in coordinating the response of cells to a diverse range of environmental stresses. Loss of p53 function is associated with loss of normal cell cycle control, diminished apoptosis, and genomic instability and is strongly associated with the neoplastic phenotype. We have a detailed knowledge of the biochemical properties of p53 and its activity as a transcription factor regulating diverse aspects of cellular function, but the in vivo physiological relevance of many of these remains uncertain. Nevertheless, p53 represents a highly significant potential target for novel therapeutic intervention; however, the further development of clinical applications and novel therapeutic strategies utilizing our knowledge of p53 is absolutely contingent upon bridging the gap between our biochemical understanding and our much less well-developed insights into the role of p53 in relevant physiological systems in vivo. PMID- 10388109 TI - Gene Transfer Technology in Therapy: Current Applications and Future Goals. AB - Gene therapy has attracted much interest since the first submissions of phase I clinical trials in the early 1990s, for the treatment of inherited genetic diseases. Preliminary results were very encouraging and prompted many investigators to submit protocols for phase I and phase II clinical trials for the treatment of inherited genetic diseases and cancer. The possible application of gene transfer technology to treat AIDS, cardiopathies, and neurologic diseases is under evaluation. Some viral vectors have already been used to deliver HIV-1 subunits to immunize volunteers who are participating in the AIDS vaccine programs in the USA. However, gene delivery systems still need to be optimized in order to achieve effective therapeutic interventions. The purpose of this review is to summarize the latest achievements in improving gene delivery systems, their current application in preclinical studies and in therapy, and the most pressing issues that must be addressed in the area of vector design. PMID- 10388110 TI - The HER-2/neu Oncogene in Breast Cancer: Prognostic Factor, Predictive Factor, and Target for Therapy. AB - The HER-2/neu oncogene encodes a transmembrane tyrosine kinase receptor with extensive homology to the epidermal growth factor receptor. HER-2/neu has been widely studied in breast cancer. In this review, the association of HER-2/neu gene and protein abnormalities studied by Southern and slot blotting, immunohistochemistry, enzyme immunoassays, and fluorescence in situ hybridization with prognosis in breast cancer is studied in depth by review of a series of 47 published studies encompassing more than 15,000 patients. The relative advantages of gene amplification assays and frozen/fresh tissue immunohistochemistry over paraffin section immunohistochemistry are discussed. The significance of HER 2/neu overexpression in ductal carcinoma in situ and the HER-2/neu status in uncommon female breast conditions and male breast cancer are also considered. The potential value of HER-2/neu status for the prediction of response to therapy in breast cancer is presented in the light of a series of recently published studies showing a range of impact on the outcome of patients treated with hormonal, cytotoxic, and radiation therapies. The evidence that HER-2/neu gene and protein abnormalities in breast cancer predict resistance to tamoxifen therapy and relative sensitivity to chemotherapy regimens including adriamycin is presented. The review will also evaluate the status of serum-based testing for circulating the HER-2/neu receptor protein and its ability to predict disease outcome and therapy response. In the final section, the review will briefly present preliminary data concerning the use of antibody-based therapies directed against the HER-2/neu protein and their potential to become a new modality for breast cancer treatment. The recently presented phase III clinical trial evidence that systemic administration of anti-HER2 antibodies (Herceptin®), alone and in combination with cytotoxic chemotherapy in patients with HER-2/neu overexpressing primary tumors, can increase the time to recurrence and overall response rates in metastatic breast cancer is reviewed. PMID- 10388111 TI - Current Management of Neuroblastoma. AB - Neuroblastoma is one of the more common pediatric cancers. Although there have undoubtedly been major advances in therapy over recent decades, there is still room for significant improvements in outcome to be made. Neuroblastoma is a disease with a variable outlook, encompassing a spectrum-from patients with disease which may undergo spontaneous regression, through those who may be cured with limited treatment, to others whose disease is refractory even to the most aggressive management strategies. Great progress has been made toward understanding the basic biology of the disease, which allows reliable allocation of individual patients to good, intermediate, or poor prognostic groups and aids selection of appropriate therapeutic approaches. Unfortunately, attempts to improve the outcome of patients with adverse prognostic factors have been less effective. This paper reviews the clinical and biological features of neuroblastoma which determine outcome, and outlines current management policies. Some experimental approaches involving translational research which seek to exploit the unique biology of neuroblastoma are discussed. Although currently unproven, such new treatments are undergoing clinical evaluation and may yet offer new hope for patients with this enigmatic disease. PMID- 10388112 TI - ASCO 1998: A Commentary. AB - The American Society of Clinical Oncology (ASCO) occupies a central place in the professional and social life of cancer specialists. In this once-a-year happening we have the opportunity to see close colleagues from the past and reflect on the state of our profession, including clinical research and the more practical aspects of our existence and survival as practitioners. Robert Mayer, the outgoing ASCO President, and the Program Chair, Margaret Shipp, did a masterful job of creating a well-organized, informative, and exciting four days in Los Angeles. ASCO is an organization in evolution. While clinical research is still its main mission, the financial and organizational aspects of cancer care occupy an increasingly important place in this meeting, as reflected in the program itself and in the dominating presence of the drug companies on the exhibit runways. Nonetheless, the quality of the scientific sessions was outstanding. PMID- 10388113 TI - 10th NCI-EORTC Symposium on New Drugs in Cancer Therapy: Dutch Treat Revisited. AB - Scientific meetings, like living organisms, tend to evolve over time. For example, the yearly ASCO meeting, which began as a small American academic oncology gathering emphasizing clinical research, has grown into a large, international forum which increasingly emphasizes best standard of care and reimbursement issues [Editor's note: see ASCO review by Chabner, Friedberg in this issue, page 263]. One meeting which has remained true to its initial mission is the NCI-EORTC Symposium on New Drugs in Cancer Therapy which occurs every two years in Amsterdam. Originally organized to help coordinate phase I cancer drug development between the United States and Europe, the meeting fills an important scientific niche. Largely because of the clinical trials standards which have been shared in this forum, phase I data from Europe are now used to launch phase II cancer trials in the United States. Today, the meeting focus-really, the meeting's challenge-remains cancer drug development in Europe and the United States. PMID- 10388114 TI - The NCI-EORTC Symposia on New Drugs in Cancer Therapy: A Brief History. AB - HISTORY OF THE SERIES: Two organizations that made a major contribution to the discovery and development of potential new anticancer agents are the NCI (United States National Cancer Institute) and the EORTC (European Organization for Research and Treatment of Cancer). A series of specialized symposia, the NCI EORTC Symposia on New Drugs in Cancer Therapy, has been dedicated to this theme. The first four symposia were held in Brussels, Belgium, and were organized by Dr. Marcel Rozencweig. In 1984, after Dr. H.M. Pinedo had established the EORTC New Drug Development Office (NDDO) in Amsterdam, the symposium moved to Amsterdam as well. All subsequent symposia were organized by the NDDO, with Dr. Pinedo as symposium President, starting with the 5th NCI-EORTC Symposium on New Drugs in Cancer Therapy in October 1986. Since then, the symposium has seen a strong growth to over 1,750 participants from 50 countries at this year's meeting. This growth has been achieved not only by increased participation from European countries, but there has been an even stronger increase in the number of participants from other continents, notably North America and the Far East. Approximately one-third of the participants were non-European. The series has achieved a truly global scale and provides an excellent forum to everyone involved in the research and development of new anticancer agents which nowadays tends to occur in a worldwide setting. PMID- 10388115 TI - New Drugs on the Horizon: Matrix Metalloproteinase Inhibitors. AB - Matrix metalloproteinases (MMPs) are a family of enzymes involved in a number of normal cellular processes, such as regulation of endometrial growth and menstruation, and abnormal processes, such as tumor growth, invasion, and metastasis. There are now 18 distinct enzymes that fall into three functional classifications based on their substrate target: collagenases which degrade fibrillar collagen; gelatinases which degrade denatured and basement membrane collagens, and stromelysins which degrade proteoglycans and glycoproteins. The MMPs can also be separated into five categories based on structural and functional similarities. PMID- 10388116 TI - Supplemental Iron: A Key to Optimizing the Response of Cancer-Related Anemia to rHuEPO? AB - About 50% of cancer patients develop anemia; this incidence rises dramatically in patients with more advanced cancer or in those receiving chemotherapy or radiation therapy. Since the late 1980s, recombinant human erythropoietin (rHuEPO) has provided a safe and effective option for treating cancer-related anemia and fatigue. However, only about 50% of patients treated with rHuEPO adequately respond to therapy. In the chronic renal failure (CRF) population, true iron deficiency is the most common cause of an inadequate response to rHuEPO. Functional iron deficiency occurs when iron cannot be provided rapidly enough to meet the demands of rHuEPO-induced erythropoiesis, despite the presence of adequate bone marrow iron stores. It is hypothesized that functional iron deficiency can also occur in cancer patients receiving rHuEPO and may account for the lack of response in a proportion of the oncology population. Studies in CRF patients have shown that the administration of i.v. iron can correct functional iron deficiency more effectively than oral iron and may improve rHuEPO response. Therefore, it is important to monitor iron status and to address either true or functional iron deficiency prior to and during rHuEPO therapy to optimize the effect of rHuEPO in cancer patients. Studies are currently under way to determine the role of i.v. iron in treating cancer-related anemia. PMID- 10388117 TI - AIDS-Related Malignancies. AB - OVERVIEW OF MALIGNANCY IN PATIENTS WITH AIDS: Despite the advent of highly active antiretroviral therapy, the incidence of human immunodeficiency virus (HIV) associated malignancies has not decreased. The United States Centers for Disease Control (CDC) has determined that Kaposi's sarcoma, non-Hodgkin's lymphoma (including primary central nervous system lymphoma), and cervical carcinoma define the acquired immune deficiency syndrome (AIDS). Some literature reports include Hodgkin's disease, anal carcinoma, lung cancer, and non-melanomatous skin cancers as ones commonly found in people infected with HIV. The oncologist is further challenged treating a patient with malignancy whose malignancy is complicated by a concurrent compromised marrow function. This article will review the clinical presentation and current treatment for the HIV-associated malignancies and selected other tumors in the HIV-infected patient. Furthermore, HIV-associated myelosuppression is a common problem in antiviral-undertreated or antiviral-resistant patients. PMID- 10388118 TI - The Goal of Cancer Treatment. AB - As clinical oncologists, our ultimate goal in treating patients with cancer is to be able to cure their disease with a combination of treatment modalities directed at the primary tumor (surgery or radiation), and potential metastases (chemotherapy). The validity of this multimodality approach to treating cancer was initially demonstrated with the successful treatment and cure of highly chemosensitive childhood cancers, such as Wilms' tumor, and these cures were only realized when adjuvant chemotherapy was included with local control measures. We attribute our treatment successes in childhood cancers to the use of cytotoxic chemotherapy, and we attribute our inability to cure many adults with more common forms of solid tumors to the ineffectiveness of chemotherapy in these diseases. Curing disease is not the goal of most pharmacological interventions in nonmalignant diseases. With the exception of antimicrobial and anticancer chemotherapy, most of the common classes of drugs are administered with the intent of controlling the disease or the symptoms caused by disease. We administer antihypertensive agents to control blood pressure, but the underlying cause of the hypertension is not cured by this therapy. If the hypertension recurs after antihypertensive therapy is stopped, we would conclude that the therapy was successful at controlling the disease. However, if a patient's tumor relapses after completing anticancer chemotherapy, the anticancer therapy would be considered to be unsuccessful. By setting lofty goals for our therapy, we increase the probability that the treatment will not meet our own and our patient's expectations. Schipper et al. [J Clin Oncol 1995;13:801-805] proposed that we abandon the "killing paradigm," which dictates that the treatment of cancer is directed toward eradication of all cancer cells, and that we adopt a "regulatory model" of cancer. This model views cancer as a maladaptive, constantly evolving process in which cancer cells differ only slightly from normal cells as a result of a few critical genetic changes that lead to dysregulation of growth. The treatment approach under this new paradigm is debulking of tumor burden with standard multimodality therapy followed by control of residual disease by "reregulation" of the remaining cancer cells. Controlling growth and spread of this residual disease would be accomplished with non cytotoxic agents which target pathways that are responsible for the dysregulation in cancer cells. We are now on the verge of having the capacity to test this new paradigm of cancer. Advances in our understanding of the pathogenesis of many common forms of cancer at a molecular level have led to a revolution in anticancer drug development. A number of new agents that target a variety of critical molecular targets, such as the farnesyl transferase inhibitors that block ras oncogene activation, the matrix metalloproteinase inhibitors that block the enzymes involved in tissue invasion and metastasis [Editor's note: please see "New Drugs on the Horizon, page 271], and the angiogenesis inhibitors that block new vessel formation in growing tumors, are now being clinically tested. These new classes of anticancer drugs are aimed at regulating or controlling cancers rather than killing them. The potential utility of targeting the critical molecular lesion in tumor cells is illustrated by the efficacy of all-trans retinoic acid in acute promyelocytic leukemia (APL). Although the capacity of all trans-retinoic acid to induce complete remissions by inducing terminal differentiation of leukemic blasts was discovered empirically, the subsequent demonstration that the pathognomonic 15:17 translocation that is present in up to 90% of cases of APL results in the production of a dysfunctional retinoid receptor appears to explain the specificity and high level of activity of retinoid therapy in this disease. This is the first example of a cancer that can be treated by specifically targeting therapy to a pathogenetic molecular lesion. Retinoids are now being used in combination with standard chemotherapy for the treatment of APL, an example of the successful application of combining a molecularly targeted agent with conventional cytotoxic chemotherapy. The development and use of molecularly targeted agents for the treatment of cancer may require us to view cancer in a new light and to adjust our goals and expectations of its treatment as well as the endpoints of our clinical trials. However, pharmacologically controlling cancer may result in an equally acceptable outcome for our patients if it leads to what Schipper et al. termed a "functional cure." PMID- 10388119 TI - Breast Cancer: High-Dose Therapy. AB - Breast cancer has become the leading indication for high-dose chemotherapy (HDC) with autologous stem cell rescue (ASCR) in North America. The rapid increase in HDC/ASCR for breast cancer has been driven by belief in the response rates and survival times demonstrated in phase II studies, which have been higher than that of historical controls. However, there is a growing body of data to suggest that selection bias has had a significant impact on the outcome of non-randomized studies of HDC. Few randomized comparisons of HDC to standard-dose chemotherapy exist. Early studies of dose-intensification and phase II studies of HDC/ASCR for patients with metastatic and high-risk disease are reviewed here, with emphasis on those studies with long-term follow-up. Studies demonstrating the selection bias present in phase II studies of HDC are also discussed. Finally, randomized studies of HDC/ASCR are reviewed, including the National Cancer Institute high priority trials. Currently, HDC/ASCR for the treatment of breast cancer can only be recommended in the context of a clinical trial. Results of several large randomized trials are awaited to determine the future of HDC/ASCR for breast cancer. PMID- 10388120 TI - Integration of Systemic Chemotherapy in the Management of Primary Breast Cancer. AB - Breast cancer cells can metastasize early in the development of primary tumors. Adjuvant chemotherapy improves disease-free survival and overall survival (OS) in patients with early-stage breast cancer, both in premenopausal and postmenopausal women. Tamoxifen improves OS in patients whose tumors are estrogen-receptor positive, regardless of age. Although the relative risk reduction with these interventions is the same for all patients, the absolute benefit is more prominent in patients at the highest risk of relapse. All patients with involved lymph nodes should receive adjuvant chemotherapy if no contraindications occur. In patients with negative lymph nodes, adjuvant chemotherapy is recommended if the invasive tumor is moderately or poorly differentiated, the tumor size is larger than 1 or 2 cm, or hormone receptors are negative. Other prognostic and predictive indicators of response to chemotherapy and hormonal therapy remain investigational. Models that combine several factors to determine a patient's risk of relapse and death are presented. Neoadjuvant chemotherapy in conjunction with radiation therapy and surgery is the treatment of choice for patients with locally advanced breast cancer. Recently, this approach has been extended to patients with small, operable tumors. Neoadjuvant chemotherapy can downstage tumors effectively, leading to improved cosmetic results and possibly to a reduction in local recurrence rate. Another advantage of neoadjuvant therapy is the in vivo assessment of tumor sensitivity to chemotherapy, which allows optimization of available therapeutic agents. One disadvantage of neoadjuvant chemotherapy is that preoperative treatment causes an alteration of information regarding lymph node status prior to systemic treatment. Although the optimal chemotherapy regimen has not been established, doxorubicin-containing regimens are considered superior to non-doxorubicin-containing regimens. The role of high dose chemotherapy is not well defined in the adjuvant setting and remains investigational. The taxanes paclitaxel and docetaxel are moving rapidly from the metastatic setting to the adjuvant setting. A better understanding of the biology of breast cancer is providing molecular tools to study novel treatment approaches. Oncogenes and tumor suppressor genes are emerging as important indicators of response to chemotherapy. These molecules, in turn, become targets for therapy. Novel agents under development are presented. PMID- 10388121 TI - Strategies for the Use of Epoetin Alfa in Breast Cancer Patients. AB - Anemia is a common complication in cancer patients undergoing chemotherapy, and its severity depends on both the type of antineoplastic drugs and the clinical status of the patient. Breast cancer patients undergoing standard chemotherapy develop clinically significant anemia in up to 25% of cases. This percentage, moreover, increases up to 63% when more intensive chemotherapy regimens are used. The therapeutic use of erythropoietin in anemic patients, i.e., in patients with hemoglobin levels below 9-10.5 g/dl, is able to correct the anemic status in nearly 40%-80% of such patients, but it does not completely eliminate the need of blood transfusions: 20%-40% of patients need to be transfused despite the erythropoietin treatment. An alternative strategy for optimizing the erythropoietin treatment is its use in the prevention of anemia, i.e., in patients with normal hemoglobin values but at high risk of becoming anemic. In a phase III study, we evaluated the role of erythropoietin in the prevention of anemia in breast cancer patients undergoing dose-intensive chemotherapy. Clinically significant anemia occurred in 52% (95% CI = 33-69) of control patients and in no patient (95% CI = 0-14) in the erythopoietin arm (p =.00001). After six cycles of chemotherapy the mean hemoglobin decrease was 3.05 g/dl (± 1.0, 95% CI = 2.6-3.5) in the control arm and 0.8 g/dl (± 1.4, 95% CI = 0.3-1.4) in the erythropoietin arm. Moreover, 6.4% of control patients needed blood transfusion compared to no patients in the erythropoietin arm. Erythropoietin is active in both the treatment and the prevention of anemia in cancer patients undergoing chemotherapy. Due to its high economic cost, efforts should be made to identify subsets of patients in whom the preventive use could be cost-effective. Patients undergoing chemotherapy associated with a high risk of anemia could benefit from preventive use of erythropoietin in special circumstances, such as presence of risk of myocardial or cerebral ischemia, uncommon blood group, or religious beliefs hindering blood transfusions. Moreover, anemia prevention could be considered in patients at high risk of requiring blood transfusions, such as patients with low baseline value of hemoglobin or with a hemoglobin decrease of >/=2 g/dl after the first cycle of chemotherapy. PMID- 10388122 TI - Hereditary Factors in Gynecologic Cancer. AB - Cancer predisposition in some families is known to be the result of germ-line mutations. The most noteworthy hereditary gynecologic cancer syndromes include hereditary breast-ovarian cancer (HBOC) syndrome, wherein BRCA1 and BRCA2 germ line mutations have been identified, and hereditary nonpolyposis colorectal cancer (HNPCC) of the Lynch syndrome II variant, wherein hMSH2, hMLH1, hPMS2, hMSH3, and hMSH6 germ-line mutations have been identified. DNA testing for specific cancer-associated germ-line mutations is now available for HBOC and HNPCC syndrome family members who are in the direct line of inheritance. Genetic counseling is mandatory prior to DNA testing and at the time of disclosure of findings. A patient found to be negative for the family's particular cancer associated germ-line mutation can revert to general population screening recommendations. When a deleterious mutation is identified, the physician is able to predict a patient's lifetime susceptibility to breast and ovarian carcinomas in the HBOC syndrome or the cancers which characterize the Lynch syndrome II variant of HNPCC, particularly carcinomas of the colon, endometrium, and ovary. Management strategies can be offered which are designed to take advantage of the natural history of that distinct hereditary cancer syndrome. We discuss the unfolding developments concerning familial and heritable susceptibilities, molecular genetics, and possible carcinogenic co-factors of the three most common gynecologic cancers: carcinomas of the uterine cervix, endometrium, and ovary. We offer rationales for management based on current epidemiologic and clinical data and emerging technologies. PMID- 10388123 TI - Carcinoid Tumors: Current Concepts in Diagnosis and Treatment. AB - Neuroendocrine tumors of the gut, carcinoids have been a diagnostic and therapeutic challenge over the years. The primary diagnostic work-up includes biochemical testing, particularly analysis of chromogranin A and urinary 5-HIAA. The most sensitive localization procedure is somatostatin receptor scintigraphy, which will be supplemented by ultrasonography for liver metastases and concomitant biopsy for histopathological verification. The treatment needs a multimodal approach, including surgery, embolization, tumor-targeted radiotherapy, and biotherapy. Chemotherapy plays a small role in the treatment of classical midgut carcinoids. Two decades ago, the median survival of patients with malignant carcinoid tumors and the carcinoid syndrome was only two years, but today, using this multimodal approach including biotherapy, it is more than eight years for the same category of patients. This may not only reflect the most effective treatment, but also a more active attitude to therapy among physicians. Future therapy will be tumor-biology-based and "tailor-made" for each patient. PMID- 10388124 TI - The Rationale for Performing Autologous Peripheral Blood Stem Cell Transplants in Community Cancer Centers. AB - There is debate over whether or not the technology of peripheral blood stem cell (PBSC) support of high-dose chemotherapy (HDC) should be disseminated to practicing oncologists or continue to be administered only in academic referral centers. High-dose therapy with stem cell support is now the standard of care for selected patients with lymphoma, multiple myeloma and possibly breast cancer. Such therapies, delivered in a clinical trials setting, need to be more widely available to eligible patients. This manuscript presents the rationale for performing HDC with PBSC support in community cancer centers. The availability of PBSC has made the delivery of well-established HDC regimens safe and effective in an outpatient setting. Delivery of such therapy where the patient lives has many economic and social advantages to the patient compared to referral to a transplant center. In addition, more patients can be treated more cost effectively if such therapy is administered locally where the patient lives. From the scientific point of view, improved access to HDC in the community should increase accrual to clinical trials, allowing the generation of outcome data more rapidly. PMID- 10388125 TI - Research and Quality Medicine in the Community: The Paradigm of Marrow Transplantation. AB - INTRODUCTION: Bone marrow transplantation (BMT), including the use of peripheral blood progenitor cells (PBPC), is a promising new technology for cancer treatment. BMT is defined here as the use of hematopoietic progenitor cells to support intensive cancer treatment. When obtained from the peripheral blood, these cells are called PBPC. Randomized trials suggest superiority of autologous BMT compared to conventional chemotherapy for patients with relapsed, responsive intermediate grade lymphoma or responding myeloma. Large phase II studies suggest benefit for selected patients with metastatic and primary breast cancer. These and additional data suggest an emerging role for BMT in cancer treatment. Counterbalancing these encouraging data, however, are the high cost and morbidity of BMT treatments. Rapid changes in technology also make it likely that major alterations in treatment delivery and cost improvements will occur within the next few years. How should society decide when this and other expensive and risky high-technology treatment should be performed in the community? The critical concepts which underlie this decision are quality and research. PMID- 10388126 TI - Schwartz Center Rounds. A Staff Dialogue on Phase I Trials: Psychosocial Issues Faced by Patients, Their Families, and Caregivers. AB - Shortly before his death in 1995, Kenneth B. Schwartz, a cancer patient at Massachusetts General Hospital, founded The Kenneth B. Schwartz Center to be housed at Massachusetts General Hospital (MGH). He created this center to advance the hopes, goals, and ideas expressed in his article, "A Patient's Story," published in the July 16, 1995 issue of the Boston Globe Magazine. The Schwartz Center is a non-profit organization dedicated to strengthening the relationship between patients and caregivers and to supporting and advancing "compassionate health care delivery in which caregivers, patients and their families relate meaningfully to one another in a way that provides hope to the patient, support to caregivers and sustenance to the healing process." One of the Center's major projects is the sponsoring of the Schwartz Center Rounds, a monthly, multidisciplinary forum in which caregivers discuss a specific cancer patient and the important psychosocial issues faced by the patient, family and caregivers. The forum allows caregivers to reflect on their experiences with patients and to gain support and insight from fellow staff members. The following case discussion was addressed at the January 1998 Schwartz Center Rounds. In this article, the case will be presented, followed by verbatim dialogue from the rounds and a subsequent discussion of the relevant issues with emphasis on staff psychosocial issues. J.T. was a 43-year-old man who developed adenocarcinoma of the lung and was treated at MGH. He died while participating in a phase I trial, resulting in marked frustration and distress among his caregivers. Staff questioned whether cancer patients entering phase I trials and their families receive unbiased information about the possible risks and benefits of the trial. They were also concerned about whether or not patients and their families really understand the physical and emotional risks of a trial. Moreover, they addressed whether patients are presented with alternatives to enrolling in a phase I trial, such as palliative care. Despite all these concerns, caregivers are reconciled to the belief that patients do value the opportunity to participate in phase I trials, in that they can contribute hope and meaning to other patients' struggles with cancer. PMID- 10388127 TI - The Importance of Planned Dose of Chemotherapy on Time: Do We Need to Change Our Clinical Practice? AB - INTRODUCTION: Dose-limiting hematological toxicity is encountered with many chemotherapy regimens; it is common clinical practice to reduce the dosage or delay the administration of the next cycle of chemotherapy in response to toxicity [1, 2]. Reducing or delaying the dose, however, will reduce the dose intensity of treatment. This practice may achieve some reduction in toxicity, but may also decrease the therapeutic effect of the treatment. This review will examine the evidence for a relationship between reductions in dose intensity of treatment and suboptimal outcome, as distinct from studies seeking to improve survival by using dose-intensified chemotherapy regimens. PMID- 10388128 TI - The 'Sentinel Node' Concept: More Questions Raised than Answers Provided? AB - The prognosis of malignant disease is essentially determined by the metastatic potential of the primary tumor. In the past, scientific attention was chiefly directed to systemic metastasis. A multitude of biological and molecular tumor markers and mechanisms has been uncovered enabling a better contemporary understanding of the process of hematogenic metastasis. This is in contrast with our knowledge of the mechanisms and pathways of lymphatic tumor spread, which is rather limited. We do know, however, that adequate surgical clearance of the regional lymphatics improves treatment results of many tumors. How far this lymph node dissection is directly therapeutic is a source of controversy. While in some instances, a stage-adjusted survival benefit was demonstrated, this may very well be attributable at least in part to the phenomenon of stage migration (Will Rodgers phenomenon) through better staging. However, it is uncontested that an established diagnosis of regional lymphatic spread is prognostically significant and should influence the indication for additional therapy and eventually for an intensive follow-up. For many tumors, the indication for adjuvant chemotherapy depends on the nodal status. On the other hand, it is equally well known that aggressive lymphatic dissection increases perioperative morbidity and even mortality. Long-term sequellae from regional lymphatic dissection are common and the effect on the local, maybe even the systemic immunological response to the malignant disease, remains unclear. To incur such risk seems especially problematic in those patients without any lymphatic spread at the time of the pathologist's work-up. Thus, there is ongoing debate about the rationale, value, extent, advantage, or disadvantage of regional surgical lymph node dissection or even radiotherapy of the regional lymphatic drainage area for many different tumors. A considerable step forward could be made if there was any diagnostic modality enabling a reliable preoperative lymph node staging. However, there is none. General criteria like size, shape, structure, or texture in variable imaging modalities are unreliable. While it is still too early to definitely evaluate in this context new diagnostic modalities like PET, immunoscintigraphy, or contrast-enhanced MRI, the initial results do not provoke clear enthusiasm toward the development of a sensitive and specific staging tool with regard to the nodal status. Adequate specificity may be obtained by external or endoluminal ultrasound-guided fine needle biopsies. However, uncertainty arises from eventually unrepresentative tissue sampling. The sentinel lymphonodectomy technique may remedy the dilemma, enabling a risk-adapted, individual indication for regional lymphatic dissection. This concept, first introduced in 1977 by Cabanas into the treatment of penis carcinoma, is based on the evidence of orderly and predictable lymphatic drainage pathways. Tumor cell progression within the lymphatic system seems to follow a sequential pattern. Primary draining lymph nodes possess the structural and functional capability to retain and to fight tumor cells efficiently. The 'sentinel node' is defined as the first tumor draining filter, and, if uninvolved, should thus adequately predict the nodal status of the disease. Skip metastases beyond an uninvolved sentinel node are supposed to be a very rare event. The reliability of the 'Cabanas approach', however, was limited by its relatively poor localization technique, and therefore failed to gain widespread acceptance. Unfortunately, the significance of the concept was not fully appreciated at the time. It is to Morton's credit that the procedure was reinstituted in malignant melanoma through a dye injection technique at the primary tumor site. This led to a rapid development and refinement of intraoperative lymphatic mapping. One major step in this process was to use radiolabeled colloids in conjunction with gamma-camera imaging or gamma probe-guided detection of the sentinel node. At present, a multitude of studies are conducted in a variety of tumors and sites, aiming at further refinements of the technique or at clinical evaluation in comparison with established lympadenectomy. The results may well change many aspects of our operative strategy in the near future. However, assuming a technically optimized procedure, will this solve the underlying tumor biological and clinical problem with respect to the necessity and efficacy of a regional lymph node dissection in node-positive cases? This is not the case; moreover, there are additional questions raised and left unanswered so far. Without any doubt, the rate of unnecessary diagnostic lymph node dissections can be considerably reduced as soon as the sentinel node concept is sufficiently validated for general use outside clinical trials. This would be a clear step forward. It is undetermined, however, how far a cancer patient with a positive sentinel node-thus already proven lymphatic metastases-would still profit from a more or less extensive lymph node dissection. It might be sufficient to use the staging information obtained through the sentinel node's status alone to decide upon adjuvant therapies. A further aspect arises from the possibility for investigating this single and supposedly most representative lymph node in far more detail than it would be possible for the large number of nodes previously sampled in conventional lymphatic dissections. This more extensive work-up may include serial sectioning, immunological and molecular techniques to enhance the sensitivity for micrometastases detection. However, very little is known about the true prognostic significance of such conventionally occult micrometastases, and even less experience exists as to the value of adjuvant therapies in those cases. Thus, while the sentinel node procedure will probably enable a more precise though less invasive lymphatic staging of malignant disease, it raises a number of important questions, as well. The general principles of multimodal treatment will have to be redefined with regard to the new diagnostic tool, which will require extensive prospective and randomized testing before a safe and reliable advantage for the patients may be established. PMID- 10388129 TI - Paclitaxel in Breast Cancer. AB - Paclitaxel has emerged as an important agent in the treatment of breast cancer. The efficacy and tolerability of this agent, as well as its lack of cross resistance with anthracyclines, have spurred intensive clinical investigation worldwide. Optimization of paclitaxel dose and scheduling and evaluation of the drug in combination regimens are a central focus of investigations. Recent clinical evidence suggests that optimal dose of single-agent paclitaxel by 3-h infusion is 175 mg/m². Trials evaluating administration schedule have not found either a 24-h or 96-h infusion to be superior to a 3-h infusion. Weekly moderate-dose paclitaxel administration is also generating much interest, given the high relative dose intensity and dose density delivered, yet very modest myelosuppression and manageable neurotoxicity observed. As first-line therapy in metastatic disease, multiple studies have documented overall response rates in the range of 30%-60%. As second-line or salvage single-agent therapy in metastatic patients, paclitaxel generally affords an overall response rate of 20% 40%, even in anthracycline-resistant patients. The novel mechanism of action and manageable toxicity of paclitaxel has led to successful incorporation into combination chemotherapy regimens. The combination of paclitaxel and doxorubicin has been the most extensively studied, with the role of this regimen continuing to evolve. Other combination regimens that appear to hold substantial promise as first-line metastatic treatment are paclitaxel with carboplatin and paclitaxel with trastuzumab (anti-HER2 antibody). The favorable results obtained in the metastatic setting have prompted phase II and phase III investigations of paclitaxel in the adjuvant and neoadjuvant settings. In the adjuvant setting, a recent phase III study has indicated that the addition of sequential paclitaxel to standard therapy affords both disease-free and overall survival benefits. Current investigations with paclitaxel will continue to optimize the role of this agent in the treatment of early- and advanced-stage breast cancer, addressing not only response rates but also survival and quality-of-life issues. The use of paclitaxel on a weekly schedule or with new therapeutic modalities, such as monoclonal antibodies, is also receiving much attention. While it is clear that paclitaxel is a very active agent in the treatment of breast cancer, it is hoped that these innovative trials will further maximize the potential of this agent in patients with breast cancer. PMID- 10388130 TI - Topotecan: An Oncologist's View. AB - Topotecan (Hycamtin(R)) is a topoisomerase I inhibitor which demonstrated a wide spectrum of antitumor activity in preclinical models. During phase I assessment, evidence of activity was most promising when topotecan was administered on an i.v. daily x 5 schedule and a dose of 1.5 mg/m(2)/day was selected for phase II/III evaluation. This regimen has been shown to have activity in a wide range of tumor types, including recurrent ovarian cancer, relapsed small cell lung cancer (SCLC), non-small cell lung cancer, colon cancer, and breast cancer, as well as hematological malignancies. In patients with ovarian cancer who had failed standard therapy, topotecan demonstrated response rates of 13% to 25%, with median times to progression of 12 to 19 weeks. Compared with paclitaxel, the response rates were similar, 20.5% and 14.0%, respectively, as were median times to progression (19 weeks for topotecan versus 15 weeks for paclitaxel). Results in recurrent SCLC have also been encouraging. Patients sensitive to previous chemotherapy have shown response rates of 19% and 39%, and even patients resistant or refractory to previous chemotherapy have had responses of 3% and 7%. Survival ranged from 20 weeks in refractory disease to 12 months in both sensitive and resistant/refractory disease combined. The safety profile of topotecan is well established. The principal toxicity is noncumulative myelosuppression, and serious sequelae are uncommon. Nonhematological toxicities are generally mild. The use of topotecan in combination regimens is promising, although clinical results are currently at an early stage. To date, topotecan has demonstrated its activity in recurrent ovarian cancer and offers a valuable addition to treatment options in relapsed SCLC. PMID- 10388131 TI - Hereditary Risk of Breast and Ovarian Carcinoma: The Role of the Oncologist. AB - The American Society of Clinical Oncology has affirmed the role of clinical oncologists in identifying and managing patients with familial cancer risk. Inherited mutations in the genes BRCA1 and BRCA2 are responsible for the majority of hereditary breast and ovarian cancers, and these mutations also increase the risk of second cancers in women already diagnosed with breast malignancy. Understanding the likelihood of breast and ovarian cancer associated with mutations in BRCA1 and BRCA2 begins with consideration of the biological basis of hereditary cancer risk. Identifying patients with hereditary risk requires documentation of appropriate family history, and recent studies have characterized criteria for identifying women most likely to have inherited mutations in these genes. Options for women with inherited mutations in BRCA1 and BRCA2 include surveillance, chemoprevention and prophylactic surgery, which must be considered separately for the management of the risk of breast cancer and of ovarian cancer. Knowledge of the hallmarks of hereditary risk, options for medical intervention, possible results of BRCA1 and BRCA2 laboratory analysis and the psychological concerns of patients about hereditary risk evaluation enables oncologists and other health care providers to effectively counsel and manage women with hereditary risk of breast and ovarian cancer. PMID- 10388132 TI - Combined Modality Treatment of Anal Carcinoma. AB - Within the framework of two phase III clinical trials, the superior results of concomitant chemotherapy and radiotherapy in the treatment of patients with locally advanced anal carcinoma were demonstrated. A further phase III clinical trial showed that the role of mitomycin C as part of the concomitant chemotherapy in combination with 5-fluorouracil appeared to be essential in obtaining a higher local control rate. All three of these randomized trials have shown that this improved local control rate results in a reduction of the colostomy rate. The standard treatment for locally advanced anal carcinoma is therefore the concomitant use of chemotherapy and radiotherapy. Future issues to be clarified include a search for the optimal radiation dose and the use of less toxic agents. For smaller tumors, the addition of chemotherapy still has to be assessed. PMID- 10388133 TI - Intermittent Androgen Suppression as a Treatment for Prostate Cancer: A Review. AB - Prostate cancer continues as the most common malignancy in men in the United States, with a large number of patients presenting with advanced disease. The current treatment for metastatic prostate cancer, permanent androgen withdrawal, is palliative. Patients treated with permanent androgen withdrawal usually relapse and die secondary to prostate cancer's ability to progress to an androgen independent state of growth. Based on preclinical studies and phase II trials, intermittent androgen suppression (IAS) appears to be a potential alternative to permanent androgen withdrawal. IAS may be a feasible alternative for the treatment of metastatic prostate cancer. In this paper, preclinical studies that form the basis of IAS and the clinical studies to date concerning IAS are reviewed. Through the cycling of reversible androgen suppression, there appears to be recovery of apoptosis and subsequent slower progression to an androgen independent state. The small clinical studies conducted to date show the feasibility of IAS, and, in a few studies, there appears to be a survival advantage over historic controls. A prospective randomized trial which is currently under way will test IAS as a treatment alternative in stage D&sub2; prostate cancer. PMID- 10388134 TI - The Prognostic Value of Thymidylate Synthase and p53 Expression in Patients Treated with Induction Chemotherapy for Squamous Cell Carcinoma of the Head and Neck. AB - Thymidylate synthase (TS) and p53 are central molecules in the regulation of cell growth. Differences in the intracellular expression of these proteins by tumor cells may have predictive value for response to chemotherapy and early failure in patients with squamous cell cancer of the head and neck (SCCHN). Immunohistochemistry was used to assess the tumor cell expression of TS and p53 in pre-therapy biopsies from patients with advanced SCCHN treated with an induction chemotherapy protocol, PFL. Samples were available from 11 of 16 nonresponders, 13 of 19 early failures with progression within 24 months of treatment, and a random selection of 13 from 45 long-term, disease-free survivors (LTS). High TS expression was seen in the majority of samples from all three groups, 67% versus 78% versus 93%, respectively; however, only one of seven (14%) samples with low TS was from a LTS patient. TS expression did not differ in patients by sex, age, site of primary tumor, differentiation or stage. p53 was expressed in 33% of patient samples and did not predict response or correlate with sex, age, site of primary tumor, differentiation, or stage. Small primary tumors with extensive nodal disease were less likely to express p53 than larger primary tumors with or without nodal involvement. The data suggest that TS and p53 content have a limited prognostic value in patients treated with PFL, although tumors with lower TS expression appeared to be less likely to respond. Differences between this study and other investigations of TS and p53 may be disease site- and regimen-specific. Statistically significant differences between response groups may emerge from larger, site-specific, protocol-driven studies of TS and p53. PMID- 10388135 TI - A Comparison of Oral Ondansetron and Intravenous Granisetron for the Prevention of Nausea and Emesis Associated with Cisplatin-Based Chemotherapy. AB - PURPOSE: To compare the efficacy and safety of oral ondansetron with i.v. granisetron each given as a single dose prior to administration of highly emetogenic cisplatin chemotherapy. PATIENTS AND METHODS: Chemotherapy-naive patients with histologically confirmed malignancies were randomized to receive a single 24 mg ondansetron hydrochloride tablet plus a 50 ml i.v. infusion of normal saline, or a single 10 ug/kg (50 ml) i.v. infusion of granisetron plus a placebo tablet in this multicenter, double-blind, parallel-group trial. Study drug was administered 30 min prior to a single i.v. infusion of cisplatin (50-75 mg/m²), given over a period of or = 8 years. The incidence increased at 12 years and peaked at 16 to 18 years of age. A majority (81.6%) of the affected children had nocturnal leg cramps 1 to 4 times per year. The mean duration of episodes was 1.7 minutes. Leg cramps were unilateral in 98.9% of cases and the ipsilateral foot also was involved in 18.9% of cases. One hundred thirty-five (73%) children had leg cramps while asleep, and the remaining 23 (12.4%) children had leg cramps in either state. Fifty-seven (30.8%) children had residual tenderness in the affected muscles. The mean duration of residual tenderness was 33.2 minutes (range: 2 minutes-1 day). We conclude that nocturnal leg cramps are common in children aged > 12 years. A majority of the affected children have leg cramps 1 to 4 times per year. The cramps are usually unilateral and occur when the children are asleep. Normal duration of the leg cramp is < 2 minutes. Residual tenderness is present in approximately 30% of the affected children. Residual tenderness, if present, usually lasts for half an hour. PMID- 10388259 TI - Migraine complicated by brachial plexopathy as displayed by MRI and MRA: aberrant subclavian artery and cervical ribs. AB - This article describes migraine without aura since childhood in a patient with bilateral cervical ribs. In addition to usual migraine triggers, symptoms were triggered by neck extension and by arm abduction and external rotation; paresthesias and pain preceded migraine triggered by arm and neck movement. Suspected thoracic outlet syndrome was confirmed by high-resolution bilateral magnetic resonance imaging (MRI) and magnetic resonance angiography (MRA) of the brachial plexus. An unsuspected aberrant right subclavian artery was compressed within the scalene triangle. The aberrant subclavian artery splayed apart the recurrent laryngeal and vagus nerves, displaced the esophagus anteriorly, and effaced the right stellate ganglia and the C8-T1 nerve roots. Scarring and fibrosis of the left scalene triangle resulted in acute angulation of the neurovascular bundle and diminished blood flow in the subclavian artery and vein. A branch of the left sympathetic ganglia was displaced as it joined the C8-T1 nerve roots. Left scalenectomy and rib resection confirmed the MRI and MRA findings; the scalene triangle contents were decompressed, and migraine symptoms subsequently resolved. PMID- 10388260 TI - A preliminary study of African-American physician involvement in the care of HIV infected patients. AB - In 1995, questionnaires were sent to the mailing list of the National Medical Association (NMA). The mail responses were supplemented by questionnaires distributed at the NMA annual meeting. Of the 709 respondents, approximately 63% were primary care providers, pediatricians, or obstetrician-gynecologists; 72% were treating from zero to 10 human immunodeficiency virus (HIV) patients while 9% were treating > 90 HIV patients; and 12% had been treating HIV patients > 10 years. The majority of these patients were African American; male-to-male sex and injecting drugs were the two major risk factors. Complexity of HIV care and lack of reimbursement were the principal barriers to providing HIV care. The burden of providing HIV care is borne by a relatively small number of physicians, and African-American physicians are actively involved in this care. Programs are needed to increase the number of African-American providers treating HIV patients and to provide appropriate reimbursement for providing this care. PMID- 10388261 TI - Preadmissions programs and enrollment of underrepresented minority students before and during successful challenges to affirmative action. AB - The association between the percent change in first-year and total underrepresented minority student enrollment and the presence of preadmission programs at Liaison Committee on Medical Education (LCME) accredited medical schools was assessed before and during successful legal and legislative challenges to affirmative action. The percent change in under-represented minority student enrollment was determined by comparing enrollment data for the academic years 1993-94 and 1996-97. Schools were categorized as having either a negative or positive percent change in their enrollment of underrepresented minority students. Logistic regression was used to determine the association of the percent change in under-represented minority student enrollment and the presence of a preadmission program while controlling for schools' financial support and the presence of postbaccalaureate programs. Fifty-six percent of the included medical schools had preadmission programs. Schools with a positive percent change were significantly more likely to have preadmission programs compared with schools with a negative percent change. There was no association between the presence of preadmission programs and the percent change in total enrollment. These results indicate that the presence of preadmission programs is positively associated with increases in first-year underrepresented minority student enrollment during the successful challenges to affirmative actions. PMID- 10388262 TI - Spontaneous pneumomediastinum in an 18-year-old black Sudanese high school student. AB - Spontaneous pneumomediastinum (SPM) is defined as pneumomediastinum in the absence of an underlying lung disease. It is the second most common cause of chest pain in young, healthy individuals (< 30 years) necessitating hospital visits. It is surpassed in frequency in this setting only by spontaneous pneumothorax. These two conditions may coexist in 18% of patients. The incidence of spontaneous pneumomediastinum varies in different communities and generally is relatively uncommon. Inhalational drug use (cocaine and cannabis) have been associated with a significant number of cases, although cases with no apparent etiologic or incriminating factors are well recognized. Also its recurrence, though uncommon, is worthy of note. It is a benign clinical condition with diverse clinical presentations. Physicians' knowledge of the presentation, treatment, and prognosis of SPM will guard against the need for expensive radiologic and laboratory tests. The differential diagnosis of chest pain, shortness of breath, and dysphagia include cardiac, pulmonary, and esophageal diseases. The tendency to pursue these entities may lead to laboratory investigations such as electrocardiograms, arterial blood gases, ventilation/perfusion scans, and contrast radiographic studies of the esophagus. PMID- 10388263 TI - Infection status of dragonflies with Plagiorchis muris metacercariae in Korea. AB - Plagiorchis muris has been found in both house and field rats as well as in humans. The infection status of the second intermediate hosts of P. muris is prerequisite in understanding their biological features in an ecosystem. Six species of dragonflies were caught in a wide range of areas in Korea; and they were Sympetrum darwinianum, S. eroticum, S. pedomontanum, S. infuscatum, Pantala flavoscens, Calopteryx atrata, and Orthetrum albistylum speciosum. The occurrence of P. muris metacercariae in dragonflies was nationwide with various infection rates. The metacercarial burden of P. muris in the surveyed areas was the highest in S. eroticum followed by S. darwinianum, S. pedomontanum, and C. atrata. The highest infection rate by P. muris metacercariae was found in S. darwinianum followed by S. eroticum. The metacercarial burden was particularly heavy in the dragonflies found in Hamyang-gun and Kosong-gun, Kyongsangnam-do. It is, therefore, likely that dragonflies play a significant role as the second intermediate host in the life cycle of P. muris in Korea. PMID- 10388264 TI - Border malaria characters of reemerging vivax malaria in the Republic of Korea. AB - Since 1993, the number of vivax malaria cases has increased every year in the northern part of the Republic of Korea (ROK). This study was designed to characterize factors related to the reemergence of malaria in the ROK. A total of 21 cases diagnosed in 1993 and 1994 distributed sporadically in the narrow zone along the demilitarized zone (DMZ). Of total 317 civilian inhabitant cases reported in 1994-1997, 287 cases were studied and 80.8% of them resided within 10 km from the southern border of the DMZ. The frequency distribution of anti Plasmodium vivax antibody titers using indirect fluorescent antibody test was compared in three villages in relation with distance from the DMZ. The number of inhabitants with high antibody titers was larger in the village nearest to the border than that in more distant villages. The present results highly suggested that the reemerging vivax malaria start in the border area, most possibly caused by infected mosquitoes which flew across the border. This pattern of transmission repeated year after year. PMID- 10388265 TI - Long-lasting sonographic and histopathological findings in cured clonorchiasis of rabbits. AB - To ascertain residual sonographic and histopathological findings of clonorchiasis after treatment, the present study evaluated sonographic findings in rabbits which were infected with 500 metacercariae of C. sinensis every 6 months for 18 months after treatment with praziquantel. The sonographic findings were analyzed in terms of intrahepatic bile duct dilatation and periductal echogenicity, and histopathological findings were observed after the last sonographic examination. Compared with the sonographic findings before treatment, dilatation of the intrahepatic bile ducts became mild to some degree in four of the seven cases and increased periductal echogenicity resolved in four of them. The histopathological specimens after 18 months showed that periductal inflammation has almost resolved but moderate dilatation of the intrahepatic ducts and mucosal hyperplasia persisted. The periductal fibrosis minimally resolved. The long-lasting sonographic findings in cured clonorchiasis make sonography less specific. PMID- 10388266 TI - Pathogenicity of Korean isolates of Acanthamoeba by observing the experimental infection and zymodemes of five isoenzymes. AB - To determine the pathogenicity of Acanthamoeba spp. isolated in Korea and to develop a isoenzymatic maker, the mortality rate of infected mice, in vitro cytotoxicity against target cells and isoenzyme band patterns were observed. Five isolates of Acanthamoeba spp. (YM-2, YM-3, YM-4, YM-5, and YM-7) were used in this study as well as three reference Acanthamoeba spp. (A. culbertsoni, A. hatchetti, and A. royreba). According to the mortality rate of infected mice, Korean isolates could be categorized into three groups high virulent (YM-4), low virulent (YM-2, YM-5, YM-7) and the nonpathogenic group (YM-3). In addition, the virulence of Acanthamoeba spp. was enhanced by brain passage in mice. In the cytotoxicity assay against chinese hamster ovary cells, especially, the cytotoxicity of brain-passaged amoebae was relatively higher than the long-term cultivated ones. The zymodeme patterns of glucose-6-phosphate dehydrogenase (G6PD), malate dehydrogenase (MDH), hexokinase (HK), glutamate oxaloacetate transaminase (GOT) and malic enzyme (ME) of Acanthamoeba spp. were different among each isolate, and also between long-term cultured amoebae and brain passaged ones. In spite of the polymorphic zymodemes, a slow band of G6PD and HK, and an intermediate band of MDH were only observed in pathogenic Acanthamoeba spp., which should be used as isoenzymatic makers. PMID- 10388267 TI - Eosinophil and IgE responses of IL-5 transgenic mice experimentally infected with Nippostrongylus brasiliensis. AB - Eosinophil and IgE responses of interleukin (IL)-5 transgenic and normal C3H/HeN mice were studied after experimental infection with Nippostrongylus brasiliensis (Nb). Intestinal worms were recovered at day 5 post-infection (PI), and numbers of total white blood cells (WBC) and eosinophils, and total serum IgE and anti hapten (dinitrophenyl) (DNP) specific IgE titers, were measured at days 0, 14 and 21 PI. IL-5 mice appeared resistant to Nb infection showing a significantly lower worm recovery rate than normal mice (P < 0.05). Total WBC and eosinophil counts (/mm3) were significantly increased in Nb infected normal mice (P < 0.05), but unchanged (total WBC) or decreased (eosinophils) in IL-5 mice at day 21 PI. The total serum IgE level remarkably increased in normal mice, but only a little in IL-5 mice at days 14 and 21 PI. Priming with DNP brought about more remarkable increases of the total and anti-DNP specific IgE in normal mice than in IL-5 mice. The results show that IL-5 mice are resistant to Nb infection, and that eosinophil and IgE responses in these mice are not augmented by Nb infection. PMID- 10388268 TI - Partial cross-resistance between Strongyloides venezuelensis and Nippostrongylus brasiliensis in rats. AB - Rats were immunized through an initial infection with 1,000 filariform larvae (L3) of Nippostrongylus brasiliensis and after complete expulsion of worms they were challenged with 1,000 L3 of Strongyloides venezuelensis to investigate whether cross-resistance developed against a heterologous parasite. Nippostrongylus brasiliensis-immunized rats developed a partial cross-resistance against S. venezuelensis migrating larvae (MSL3) in the lungs and adult worms in the small intestine. The population of MSL3 in the lungs were significantly lower (P < 0.05) in immunized rats (22.0 +/- 7.4) compared with controls (105.0 +/- 27.6). The populations of adult worms, egg output and fecundity were initially decreased but from day 14 post-challenge they did not show any significant difference between immunized and control rats. However, the length of worm in immunized rat was revealed as retardation. Peripheral blood eosinophilia was significantly decreased (P < 0.05) on day 7 post-challenge and then gradually increased, which peaked on day 42 post-challenge when most of the worms were expelled. These results suggest that peripheral blood eosinophilia is strongly involved in the worm establishment and expulsion mechanisms. PMID- 10388269 TI - Variation of antigenicity and serological reaction to Pneumocystis carinii in Korea. AB - The present study observed the variation of antigenicity of Pneumocystis carinii and serum IgG antibody reaction to the antigens from different localities in Korea. Antigens of rat P. carinii and sera of inhabitants were collected at Chunchon. Chungju, Kwangju, and Seoul during 1995-1996. Enzyme-linked Immunosorbent Assay and immunoblot were used for immune reaction. Absorbance of 1,294 human sera ranged between 0.01 and 0.93. Sera from Chunchon showed higher absorbances than those from other areas. Immunoblotting revealed IgG antibody reactions to 116, 100, and 45-55 kDa antigenic bands of rat P. carinii, but the frequencies of positive reaction to individual bands were variable by localities. Total 62.6% of the sera showed the reaction to 116 kDa band while 37.7% reacted to 100 kDa band and 32.0% to 45-55 kDa bands. For the reaction to 116 kDa, the reaction rate was 60.0% to 82.6% by localities. It is found that the reaction rates of the human sera to rat P. carinii antigen are variable according to the localities. Also, the high molecular antigen of 116 kDa of rat P. carinii is the most frequent antigenic band reacting to human sera. PMID- 10388270 TI - A case of Strongyloides stercoralis infection. AB - Strongyloidiasis has been recognized as one of the life-threatening parasitic infections in the immunocompromised patients. We report an intestinal infection case of Strongyloides stercoralis in a 61-year-old man. Rhabditiform larvae were detected in the stool examination and developed to filariform larvae having a notched tail through the Harada-Mori filter paper culture. The patient received five courses of albendazole therapy but not cured of strongyloidiasis. PMID- 10388271 TI - Axenic cultivation and characterization of Giardia lamblia isolated from humans in Korea. AB - Inoculating of human fecal cysts to suckling Mongolian gerbils, two Giardia lamblia isolates, K1 and K2, were established as axenic cultures. Using this in vitro culture, both isolates were characterized as a "medium-rate grower" upon its growth pattern. These two Giardia isolates were grouped by using two genetic analysis. With genetic analysis of SSU-rDNA sequences, both K1 and K2 were found as members of Hopkins' group 1, despite some nucleotide differences noticed in K2 (5 differences/292 bases). The other genetic study used PCR-RFLP of the tim (triose phosphate isomerase) gene. Both of K1 and K2 were found to belong to Nash's group 2. Our results suggest that Nash's group 2 can not be a separate group, but a part of Hopkins' group 1. PMID- 10388272 TI - [Quality development in health care--time for strategic choices! Open account challenging but inevitable]. PMID- 10388274 TI - [Considerations about sick-listing]. PMID- 10388273 TI - ["Negligence" in connection with incidents?]. PMID- 10388275 TI - [Quality of life--a useful measure of quality of health care]. PMID- 10388276 TI - [Drug therapy of obesity will come later]. PMID- 10388277 TI - [Linne was right in many things]. PMID- 10388278 TI - [Make the medical risk management a subject in medical training]. PMID- 10388279 TI - [Cardiovascular treatment potentials. P2 receptors important for future drugs]. AB - Several receptors activated by extracellular nucleotides (ATP, ADP, UTP and UDP) have recently been cloned. These P2 receptors mediate a multitude of cardiovascular effects such as positive inotropic effects in the heart, platelet aggregation, release of endothelial factors, growth stimulation of vascular smooth muscle cells, vasomotor effects and blood pressure regulation. The physiological and pathophysiological importance of P2 receptors is established, and the first P2 receptor antagonists (ticlopidine and clopidrogel) are already in clinical use as inhibitors of platelet aggregation in the prevention of ischaemic heart disease and stroke. The development of selective antagonists for other P2 receptor subtypes will lead to better understanding of cardiovascular disease processes and yield new therapeutic options. PMID- 10388280 TI - [Dyspepsia--can we follow dogmatic guidelines in a nuanced reality? Individual management of patients with dyspepsia with risk of stomach ulcer]. AB - A review of subject literature suggests management strategies for the treatment of dyspepsia to be characterised by marked differences. In some strategies the possibility of indirect Helicobacter pylori testing (e g, breath tests) is ignored and endoscopy recommended as the only appropriate investigation; only those with verified disease are treated, and the risk of antibiotic overuse is minimised. In other strategies, indirect H pylori testing is recommended for younger patients (< 45 years of age) without alarming symptoms, either to select patients for endoscopy or to eradicate the infection (i.e., irrespective of whether the patient has peptic ulcer disease or not, on the grounds that the risk of having or developing such disease is thus minimised. The article consists in discussion of the different strategies, and suggests a choice of investigations tailored to the needs of the individual patient to be preferable to dogmatic adoption of either approach, though endoscopy should be available without delay if required. Moreover, the accuracy of the various indirect H pylori tests needs to be considered. To be acceptable for use in primary care, it is suggested that recommendations regarding interventions for the various disorders associated with dyspepsia should be characterised by reasonably comparable risk levels. An algorithm with alternative strategies suited to available facilities and the patient's needs and wishes is also presented. PMID- 10388281 TI - [LGL syndrome can imitate Felty's syndrome. The diagnosis can be established by a simple test]. AB - The article consists in a discussion of neutropenia caused by large granular lymphocytes (LGLs), illustrated by a review of the literature and case reports of five patients with LGL syndrome and one patient whose clinical characteristics were more consistent with classic Felty's syndrome. Recent years have witnessed advances in our knowledge of clonal expansions of suppressor-type T-cells and their capacity to induce neutropenia. The phenotypes of such cells are CD3+, CD8+ and CD57+. The syndrome is often seen in patients with rheumatoid arthritis, and if they also manifest splenomegaly it may be confused with Felty's syndrome. Appropriate evaluation and treatment of the condition are also discussed, and an attempt made to clarify the confusing terminology. PMID- 10388282 TI - [Fulminant acute pancreatitis caused by a large parathyroid adenoma. Hyperparathyroidism was diagnosed after 5 years]. PMID- 10388283 TI - [A case report: mesenterial vein thrombosis behind diffuse abdominal symptoms]. PMID- 10388284 TI - [Nicotine replacement always better than further smoking or using snuff]. PMID- 10388285 TI - [Management of PTSD among Bosnia refugees. Competent personnel when it comes to language and culture--an important resource]. PMID- 10388286 TI - [A reply about milk porridge. Infant food is also a question of nutritional physiology]. PMID- 10388287 TI - [Clarence Crafoord--one of the great pioneer surgeons of the century]. PMID- 10388288 TI - [Apropos of unsolicited medical intervention....]. PMID- 10388289 TI - [Students do not return home to Tanzania after completing their medical training. Barefoot physicians have to work in the countryside]. PMID- 10388290 TI - [Early surgery in small abdominal aortic aneurysm? Guidelines based on an English study]. PMID- 10388291 TI - [Riks-Stroke with data on stroke shows the benefit of quality registries. Regional initiatives can make it even better]. PMID- 10388292 TI - [The mouth is a part of the body--yes!]. PMID- 10388293 TI - [Has the professor lost contact with reality?]. PMID- 10388295 TI - [Increasing drug costs--physicians' responsibility?]. PMID- 10388294 TI - [Does physical exercise during pregnancy prevent permanent weight gain?]. PMID- 10388296 TI - [A pseudo-debate that diverts the focus from important research]. PMID- 10388297 TI - [Spiral computed tomography--solution to diagnostic problems in pulmonary embolism?]. AB - Clinical diagnosis of pulmonary embolism is difficult and often dependent on radiological methods. In Sweden scintigraphy has hitherto been the method most commonly used, though all too often it leaves the diagnosis in doubt. Spiral computed tomography, performed during continuous infusion of contrast medium, clearly depicts the pulmonaries arteries, emboli appearing as filling defects. The examination takes less than one minute. Although the method has yet to be fully evaluated, it is already available at most Swedish hospitals. Results presented so far suggest that the technique has great potential, and may become the method of choice in the diagnosis of pulmonary embolism. PMID- 10388298 TI - [Stroke unit care saves lives. The Swedish national quality assessment registry of stroke care is the first of its kind in the world]. AB - Meta-analyses of randomised trials of acute stroke treated in specialised stroke units have yielded convincing evidence of benefits in terms of reduced mortality rates, as compared with treatment in a general ward. However, no studies had been performed to ascertain whether the promising results could be reproduced in routine clinical practice. Accordingly, a comparison of routine care of acute stroke patients in stroke units (SUs) with that in general wards (GWs) was made on the basis of data for the 14,300 cases of acute stroke from 87 units in 80 Swedish hospitals registered in 1996 at the Swedish national stroke registry, the first of its kind in the world. Among patients capable of independent daily life and fully conscious at admission, the mortality rate was lower in the SU than in the GW subgroup, both at discharge from hospital and three months after the stroke event; and three months after stroke, a greater proportion of SU patients had been discharged to their homes, and a smaller proportion were in long-term care. However, no such subgroup differences were found among patients with impaired consciousness at admission. Thus, the promising results of the randomised trials of SU treatment would appear to be reproducible in routine clinical practice, though the beneficial effect is smaller in magnitude. PMID- 10388299 TI - [New article series: the first call duty--the first, difficult experiences]. PMID- 10388300 TI - [Difficult for a young colleague--the second call should have come]. PMID- 10388301 TI - [A study in eight counties: can closure of infrequently used emergency service improve quality of care?]. PMID- 10388302 TI - [Unchanged life situation of the mentally ill in southern Bohus county]. PMID- 10388305 TI - [Food as complement to drugs. Special diet with cereals for relief in inflammatory bowel disease]. PMID- 10388303 TI - [Experiences from a center for asylum-seeking persons: consultation is necessary to be able to cope]. PMID- 10388304 TI - [Modified cost-benefit analysis takes equity into consideration. Treatment of brain tumors is more cost-effective than hip replacement]. AB - As health care resources are held to be insufficient to permit free choice of available treatment options, prioritizing is necessary. A precondition of careful prioritizing is comprehensive knowledge of the consequences, both for the patient and the community, of adopting each of the available options in a given case. A tool used in setting priorities is cost-benefit analysis, based on the utilitarian principle of health maximisation. However, it is evident from findings in recent studies that equity is widely considered a factor which ought to be included in the analysis. The article presents a method for correcting the variable, quality-adjusted life years (QALY), by introducing an equity factor, yielding a new variable, equity-adjusted QALY (EQALY). Use of the method is illustrated by comparison of the cost-effectiveness of two procedures, hip replacement and the treatment of malignant glioma, and showing how priority ranking of the two procedures is reversed if the EQALY variable is used instead of QALY. PMID- 10388306 TI - [Radiographic diagnosis of cardiogenic pulmonary edema]. AB - Development of pulmonary edema (increased extravascular lung water) is a common and sometimes life-threatening clinical problem in critical-care unit patients. There are three principal causes: cardiac failure, overhydration, and increased pulmonary capillary permeability. Among these, cardiogenic edema consists of left heart failure and overhydration. Determining the specific cause of any given case of pulmonary edema is important and leads to more rapid and definitive treatment. A plain chest film can often explicate the cause of edema with a high degree of accuracy if careful attention is given to certain radiographic features. The principal features useful for correctly determining the cause of edema in a high percentage of cases are the distribution of pulmonary blood flow, distribution of pulmonary edema, and vascular pedicle width. Ancillary features are pulmonary blood volume, bronchial cuffing, septal lines, pleural effusion, and air bronchograms. Cardiac size and shape as well as specific intracardiac calcifications could also help distinguish cardiogenic from noncardiogenic pulmonary edema. PMID- 10388307 TI - [CT diagnosis of solitary pulmonary nodule]. AB - The CT (including HRCT) findings of solitary pulmonary nodule (SPN) were reviewed. CT currently is the imaging modality of choice for the evaluation of SPN. Important roles of CT are detecting pulmonary nodules and distinguishing malignant nodules from other benign tumors or inflammatory masses. To differentiate malignancy from benignancy, it is necessary to evaluate the CT findings of SPN, including morphology using HRCT, attenuation of the nodules using thin-section CT, and enhancement effect on contrast-enhanced CT. Also important in this distinction is the evaluation of satellite lesions around SPN and the relationships between bronchus, artery, vein, pleura, and interlobular septum. Spiral CT has greatly expanded the usefulness of CT in the evaluation of SPN and has become the imaging modality of choice for SPN by combining the advantages of a single breath-hold acquisition and improved MPR and three dimensional reconstruction capabilities. MPR and three-dimensional images of spiral CT can also be used to display the three-dimensional relationship between SPNs and bronchus, vessels, or pleura. PMID- 10388308 TI - [Phyllodes tumor of the breast with cystic portion: MR imaging with histopathological correlation]. AB - The purpose of this study was to reassess the MR imaging appearance and significance of dynamic MR imaging in phyllodes tumor with cystic portion. MR imaging in four patients with surgically proven phyllodes tumor was reviewed both radiologically and histopathologically. In all cases, T2WI showed inhomogeneous signal intensity with a hypointense area and internal septation in the solid portion. At histopathological examination, collagenous fibers and hemorrhage were seen in the area corresponding to hypointensity in the solid portion on T2WI. In the dynamic study, three cases showed the gradual type and one the rapid type. The percentage of cystic portion in the gradual type was over 30%, and in the rapid type, 6%. Cases of the gradual type, histopathologically, showed apparent hemorrhage, necrosis, cystic dilatation of the duct, sclerotic change, and a less densely stromal component than the rapid type. In conclusion, we speculate that the dynamic pattern may reflect changes in tumor angiogenesis that do not correlate with either malignancy or benignancy. On T2WI, internal septation and a hypointense area are considered useful for diagnosis. PMID- 10388309 TI - [Optimal dosage of contrast material in brain enhanced CT]. AB - PURPOSE: To assess the optimal contrast dosage for brain enhanced CT. MATERIALS AND METHODS: A total of 150 patients were randomly assigned to two groups. The patients were also divided by body weight into three groups (< 50 kg, 50-59.9 kg and > or = 60 kg). A total of 100 ml (in 77 patients) or 50 ml (in 73 patients) of contrast material (iopamidol 300 mgI/ml) was intravenously administrated at a speed of 1 ml/sec. Three diagnostic radiologists evaluated image quality by five- or two-stage scoring systems with special attention given to enhancement of the anterior and middle cerebral arteries. CT values of the basilar artery were also measured. RESULTS: In visual evaluation by the five-stage scoring system, image quality with 100 ml of contrast material was better than that with 50 ml in all body-weight groups, although no difference was observed with the two-stage scoring system in patients weighing less than 60 kg. CT values of the basilar artery were higher in patients given 100 ml than in those given 50 ml regardless of weight groups. CONCLUSION: The image quality of brain CT using 50 ml of contrast material is sufficient for correct diagnosis in patients with a body weight of less than 60 kg. PMID- 10388310 TI - [Usefulness of tumor size on MR imaging in assessing the prognosis of uterine cervical cancer treated with radiation]. AB - The purpose of this study was to assess the usefulness of MR imaging (MRI) in evaluating the primary tumor and predicting the prognosis after radiotherapy for uterine cervical carcinoma. MRI was performed before radiotherapy in 25 patients with squamous cell carcinoma. According to the staging of FIGO, 3 patients were classified as stage Ib, 1 as IIa, 5 as IIb, 13 as IIIb, 2 as IVa, and 1 as IVb. Three-dimensional diameters (transverse, anteroposterior, and craniocaudal) of the primary tumor were evaluated on T2-weighted images. For patients with tumors < or = 4 cm in craniocaudal diameter, i.e., the length of the tumor parallel to the long axis of uterine body, five-year disease-free survival (DFS) was 70%. For patients with tumors > 4 cm in craniocaudal diameter, the 5-year DFS was 37%. The difference between the two groups was statistically significant. For patients with tumors < or = 4 cm and > 4 cm in transverse or anteroposterior diameter, 5 year DFS was 63% and 50% respectively. There was no statistically significant difference between the two groups. The results showed the craniocaudal diameter of the tumor to be the most critical factor in predicting prognosis after radiation therapy in uterine cervical cancer. Moreover, MRI was an important means of evaluating the depth of uterine cervical carcinoma. In conclusion, MRI is useful in evaluating the effect of radiotherapy and predicting prognosis in uterine cervical cancer. PMID- 10388311 TI - [Natural history of intracranial meningioma after radiotherapy]. AB - The author examined the natural history of intracranial meningioma after radiotherapy using CT or MR imaging. Twenty patients with intracranial meningioma received radiotherapy from a high-energy linear accelerator (4-10 MV X rays) from 1980 to 1996. The total doses were 50 Gy to the tumor bed in single doses of 2 Gy in 5 weekly fractions. Meningiomas in 10 of 20 patients were reduced within 1 to 38 months after radiotherapy, the average being 11 months. The tumors were controlled for a median of 60 months after radiotherapy (range 19-126 months). Four other patients have shown no change in tumor size after radiotherapy. The tumors were controlled for a median of 70 months after radiotherapy (range 37-127 months). The other six patients have shown tumor growth within 3 to 25 months after radiotherapy, after which the tumors stopped growing for a median of 71 months (range 2-181 months). Neither tumor size nor histological type was related to response. The growth of tumors was controlled by radiotherapy for a median duration of 43 months in the meningothelial type, 52 months in the fibroblastic type, and 61 months in the transitional type. The median duration for all benign tumors was 52 months. A moderate correlation was noted between tumor response and functional outcome after radiotherapy in 9 patients with neurological deficits. The natural histories of intracranial meningiomas after radiotherapy were grouped into three categories. Some tumors showed no change in size over a long period. This was a characteristic response after radiotherapy that differed from that of other brain tumors. The results of this study provide important information for the follow-up of intracranial meningiomas after radiotherapy. PMID- 10388312 TI - [Quantitative study of 99mTc-Technegas SPECT for ventilatory impairment in pulmonary emphysema]. AB - 99mTc-Technegas scintigraphy is used to evaluate ventilation abnormalities in patients with pulmonary emphysema. Although abnormalities of ventilation distribution are easy to find, no objective index exits. Evaluation is subjective and differs with each radiologist. Thus, it is difficult to compare cases and the clinical course in the same case. The present study for quantitative evaluation demonstrated an excellent correlations between mean voxel values of the lung and stage classification. Furthermore, a correlation was observed between the mean and FEV1.0%. These findings indicate that the quantitative analysis of SPECT data is useful for classifying clinical stage and comparing cases. PMID- 10388313 TI - [Uptake of 201TlCl and 99mTc-tetrofosmin in neurofibroma: a case report]. AB - A 34-year-old man with neurofibromatosis type 1 (von Recklinghausen's disease) was examined by 201TlCl and 99mTc-tetrofosmin scintigraphies. Hypervascularity was observed in the largest tumor of the right thigh. The tumor was also depicted on both scintigrams. Tumorectomy was performed and the pathologic examination revealed it to be a neurofibroma measuring 10 x 7 x 4 cm in size. These findings suggested that 201TlCl and 99mTc-tetrofosmin scintigraphies generally were unable to distinguish malignant from benign neurofibromas with certainty. PMID- 10388314 TI - [Labyrinthine fistulas in cholesteatoma]. AB - The purpose of the present study was to investigate the clinical features of cases of cholesteatoma with labyrinthine fistulas, and in particular the pre and post-operative bone-conduction (BC). Cholestatoma patients with bone erosion or a defect found in the first stage operation were analyzed. The operations were conducted between 1992 and 1996. The patients were classified into four types, I, IIa, IIb, and III, according to Dornhoffer and Milewski's classification, which is based on different stages in the bone defect. A type I fistula is an erosion of the bony labyrinth with an intact endosteum. Type IIa is accompanied by an opened perilymphatic space with undisturbed perilymph while type IIb has a disturbed perilymph. A Type III fistula is an opened perilymphatic space with a disturbance of the underlying membranous labyrinth. Only 24 patients with type II and type III fistulas were included in this study. The location of the fistulas was the semicircular canals (SCCs) or/and the vestibula in 21 patients and in the cochlea in 3 patients. We examined the fistula by high-resolution computed tomography scan (CT scan) with 1 mm slice and 1 mm width axial-sections in 14 patients. A bone defect in the labyrinth was detected in 10 cases (71.5%) pre operatively. Pre-operative BC was worse in the patients with cochlear fistulas than in those with fistulas located in SCCs or the vestibula. Within this latter group there were 13 type IIa (group IIa), patients and 8 type IIb or III (group IIb or III) patients. However, there was no difference in the pre-operative BC between these two sub-groups Tympanoplasty was conducted in all 24 patients. The postoperative BC of group IIa and group IIb or III were compared. Two of the 13 patients in group II a (15.4%) and 3 of 8 in group IIb or III (37.5%) had a deteriorated postoperative BC. Statistical analysis revealed that the postoperative BC was more inclined to become worse in patients with advanced stage IIb or III fistulas. PMID- 10388316 TI - [A study of the movement of the articulatory organs in Japanese geminate production ANX-ray microbeam analysis]. AB - The pattern of movements of the articulatory organs, particularly the tongue and lips, in the production of Japanese geminate was analyzed using an X-ray microbeam system. Special attention was paid to clarify the difference in the pattern of movements in geminate production from that in simple and long vowel production. The subject was a Japanese male who spoke the Tokyo dialect of Japanese. Goldpellets were attached to the uppersurface of the tongue body and dorsum, lowerlip and lower Jaw using dental adhesive. Another reference pellet was placed on the nasal dorsal, and a goldfilling in the left first upper incisor was also used as a reference. The movement of each pellet during the production of test utterances was tracked using an X-ray microbeam system developed at Wisconsin University and the data output was read into a computer memory core for further analysis. During recording sessions, the subject uttered a series of nonsense test words/papiH/./paHpiH/ and /paQpiH/, where /Q/ indicates a geminate and /H/ indicates a long vowel. A comparison was also made between the utterances produced at the normal speed and those produced at the maximal speed. The movement data were analyzed using a Sun computer system. It was found that the velocity of vertical lip movement was almost constant, regardless of the type and speed of utterance and that the variation in its movement pattern was relatively small. On the other hand, the movement of the tongue body and dorsum was significantly slower and its pattern was more variable in the production of geminate compared with simple and long vowel production. It was concluded that the tongue body and dorsum movements during Japanese geminate production can be regarded as a feature-specified articulator representation called phonological underspecification. PMID- 10388315 TI - [Loss of heterozygosity of 3p21 and 9p21 in head and neck squamous cell carcinomas and its prognostic implication]. AB - To examine whether genetic factors influence the prognosis of cancer patients, several microsatellite markers were used to determine the allelic loss of certain areas of the genome. Three microsatellite markers, D3S1067, IFNA and D9S171 were used to study the loss of heterozygosity (LOH) of 3p21 and 9p21 in 93 head and neck squamous cell carcinomas. Of 57 informative cases, LOH was detected in 27 of 57 (47%) DNA samples obtained from cancer specimens when at least one marker was used. The frequency of LOH was not correlated with the clinical factors. However, the frequency of LOH was significantly higher in the recurrent cases than in the non-recurrent cases, and patients with 3p21 and/or 9p21 LOH tended to survive for a shorter period of time. These results suggested that the allelic loss at 3p21 and/or 9p21 could be correlated with the prognosis of the patients, and that it was a novel prognostic factor independent of other clinical factors concerning head and neck cancers. LOH at 3p21 and/or 9p21 may help to identify head and neck cancer patients with a poor prognosis, who need an intensive postoperative follow up protocol, or who are suitable for novel investigational therapeutic approaches. PMID- 10388317 TI - [The ratio of cervical subacute necrotizing lymphadenitis occupying superficial lymphadenopathy and its clinical findings]. AB - INTRODUCTION: Diseases in which cervical lymphadenopathy is a chief complaint are commonly observed. These cases are associated with a good prognosis, high fever and pain which usually recovers without medication. This condition is referred to as subacute necrotizing lymphadenitis (SNL). We investigated cases of SNL that were correctly diagnosed by biopsy. OBJECT AND METHODS: We examined cases of SNL that were correctly diagnosed by biopsy in the Naha Prefectural hospital between April 1987 and March 1997. We statistically analyzed the ratio of occurrence and clinical findings (age, sex, season of occurrence, physical characteristics, clinical progress, blood findings, therapy, and prognosis). RESULTS: In a total of 629 cases, a biopsy specimen from the body surface of the area affected by lymphadenopathy was obtained. Among these cases, SNL was diagnosed in 54 accounting for 9% of the total body surface biopsies and 13% of the cervical lymphadenopathic biopsies. Sex: Of the 54 subjects, 18 were males and 36 females. AGE: Most of the subjects (87%) ranged from 10 to 30 years of age. Season of occurrence: The number SNL cases decreased from 1993. Many cases occurred in the cold season, from October to March. CLINICAL FINDINGS: Forty cases were investigated as fever, swelling, pain and complications. FEVER: Six cases (15%) were not associated with fever and 34 cases (85%) exhibited fever. Swelling: All subjects demonstrated swelling for at least one week and the longest duration of swelling was six months. Swelling continued for two to three months on average. Thirty-five cases (88%) showed swelling on one side only, left or right, and five cases (12%) showed swelling on both sides. PAIN: Ten cases (25%) were without pain and 30 cases (75%) with some pain. COMPLICATIONS: Twelve cases (30%) had complications including six of drug allergy, four of dermatitis, and some cases of diabetes mellitus and hyperthyroidism. Eleven of 54 cases (20%) were admitted to the hospital. Blood findings: The white blood cell level decreased in 30 of 37 cases (82%). As shown below, increased levels of CRP (6/34), ESR (4/31) and LDH (17/31) were observed. THERAPY: Steroids were administered in 24 of 36 cases and were effective in all cases. Antibiotics were administered in 25 of 31 cases and were effective in six cases (19%)) and ineffective in 10 cases (32%). The condition in nine cases (29%) worsened. Pain killers were employed in 26 of 32 cases. They were effective 18 cases (56%) and ineffective in eight cases (25%). No subjects died. The prognoses were good and all patients recovered without sequela. CONCLUSION: SNL was detected in a large number of patients with cervical lymphadenopathy who visited our hospital, if patients who were not diagnosed correctly by biopsy were included. Many patients exhibited lymphadenopathy on one side (88%). This result was slightly higher than that previously reported. SNL is considered to be related to allergy or upper respiratory infections. This disease often occurs in cold seasons and patients often exhibit complications such as drug allergies or antoimmune diseases. PMID- 10388318 TI - [Influence of proprioceptive input from leg, thigh, trunk and neck muscles on the equilibrium of standing]. AB - To investigate and compare the roles of proprioceptive input from the leg, thigh, trunk and neck muscles on equilibrium, we performed static posturography in 50 normal subjects in the standing position on a force platform by applying vibratory stimulations to the muscles. The length of the displacement of the center of gravity, maximum sway length and sway area were measured. The amplitude of the body sway was maximum when the stimulation was applied to the dorsal neck. The forward shift of the center of gravity was also marked by stimulation applied to the dorsal neck. The amplitude of the body sway on stimulation of the leg muscles was also marked, although less than that of dorsal neck stimulation. The backward shift during stimulation of the gastrocnemius and the forward shift during stimulation of the anterior tibialis were remarkable. The results indicate that the leg muscles, which directly regulate the movement of the ankle joint, and the dorsal neck muscles, which change the static equilibrium through the central nervous system, are important for maintaining the standing posture. PMID- 10388319 TI - [Effect of inflation of the eustachian tube on tinnitus]. AB - The purposes of this study were to evaluate the effect of inflating the Eustachian tube in patients with tinnitus, and to identify diseases in which tubal inflation is indicated. Fifty-four ears of as many patients complaining of tinnitus were examined by pure tone audiometry and decreases in pitch and loudness associated with tinnitus were also evaluated by tinnitus audiometry (Danac-100, DanaJapan). Tinnitus was associated with sensorineural hearing loss (SNHL) in 44 ears and not associated in the remaining 10. The subjects were classified further into two groups: the higher tone group showed a pitch range with tinnitus of 1000 Hz or higher, and the lower tone group showed a pitch range of less than 1000 Hz. In the higher tone tinnitus group with SNLH, the tubal inflation was effective in 3 of 31 ears (10%), and in the lower tone group, 10 of 13 ears (77%). On the other hand, in the tinnitus group with no hearing loss, the method relieved tinnitus in 6 of 10 ears (60%). In the higher tone tinnitus group with no hearing loss, the tubal inflation was effective in 3 of 6 ears (50%), and in the lower tone group, 3 of 4 ears (75%). In this study, however, no ears were permanently relieved of tinnitus with tubal inflation. In the higher tone group, the duration of the reduced tinnitus was less than 10 minutes. In 69% of the lower tone group, the reduction was from 20 minutes to 2 hours. The effect did not continue for more than 2 hours at the longest. In conclusion, Eustachian tubal inflation is indicated in diseases with tinnitus as follows: 1. Lower tone tinnitus with SNHL, particularly in Meniere's disease and acute onset low-tone type SNHL, may be temporarily relieved with tubal inflation. 2. Tinnitus in an ear without SNHL that may gain transitory relief from ringing with the tubal inflation. PMID- 10388320 TI - [Report of two rare cases of fungal sinusitis]. AB - Two cases of aspergillosis of the paranasal sinuses are reported. The first case was a 30-year-old man who had a 5-month history of bilateral proptosis. Physical examination revealed nasal polyps in both middle meatus. A skin test for Aspergillus was positive. Laboratory study showed levels of serum IgE and IgE specific for Aspergillus level to be elevated significantly. Computed tomography (CT) and magnetic resonance imaging (MRI) showed pansinusitis with some bone erosion. The patient underwent bilateral Caldwell-Luc procedures and external sinus surgery (frontal, ethmoid and sphenoid sinuses). Histopathological examination showed thin septate hyphae in allergic mucin. The patient is now being treated with sinus irrigation and oral administration of fluconazole and suplatast tosilate. The second case was a 78-year-old man who had a 2-month history of nasal obstruction and a 3-week history of headaches. He also had a history of diabetes mellitus. Physical examination showed swelling of the nasal septum due to abscess. CT showed an abscess in the nasal septum and opacification of the left sphenoid sinus. There was no bone destruction. The patient underwent left sphenoid sinus surgery, and histopathological examination revealed aspergillosis of the sphenoid sinus. He presented with left visual disturbance and blepharoptosis 2 months after surgery. Ocoulusion of the internal carotid artery was revealed by MR angiography and it was thought to be caused by intracranial invasion of aspergillus. Loss of consciousness and right hemiplegia ensued despite antifungal chemotherapy. The patient died about 1 year after the onset of symptoms. Case 1 was thought to involve allergic aspergillus sinusitis, and Case 2 invasive aspergillus sinusitis. We emphasize the significance of headache, diabetes mellitus and lesion in the sphenoid sinus as a sigh of intracranial aspergillus invasion, based on our experience as well as findings reported by other clinicians in the Japanese literature. PMID- 10388321 TI - [Recent advances in geriatric rehabilitation]. PMID- 10388322 TI - [Receptors and signal transduction for the TGF-beta related factors]. PMID- 10388324 TI - [Drug therapy in elderly patients and patient's compliance]. PMID- 10388323 TI - [Clinical pharmacokinetics and pharmacodynamics in the elderly]. PMID- 10388325 TI - [Notice and contrivance on multiple drug prescription for elderly patients]. PMID- 10388326 TI - [Adverse drug reaction in the elderly]. PMID- 10388327 TI - [Quality of life in the pharmacologically treated elderly patients]. AB - The quality of life has been shown to decline with advancing age, being affected by health status, physical symptoms due to multiple chronic diseases, and the level of remaining activities of daily living of the elderly. In the pharmacologic treatment of the elderly, attention must be paid to such backgrounds and adverse side effects of drugs must be considered in order to maintain the QOL of the elderly which has already been jeopardized. Therefore, prescriptions should be consist of drugs concerning the effects on the QOL have been established. Physicians and medical personnels treating or caring for the elderly should take into account the balance between the quantity and the quality of their remaining life. PMID- 10388328 TI - [Relationships among health-promoting activities, going out and perceived transportation problems of elderly people living in a small town far from the nearest train station]. AB - To estimate the change in health-promoting activities among elderly people affected by community organizing environments, we examined the relationships among health-promoting activity, going out and perceived transportation problems. A questionnaire was sent to 567 men and women aged 60 years old and over living in a small town in Kanagawa prefecture between July 27 and August 12 in 1995. The questionnaire consisted of 42 items concerning health, social ability of daily living (including the desire to participate in social activities), attitude toward health-promoting activities, and perceived transportation problems. A total of 397 people responded and the answers from 368 people were analyzed after excluding responses from those unable to go out by themselves and those who seldom went out. Single regression analysis and multiregression analysis were used with the sum of responses for each question representing factors related to health-promoting behavior. A probability level of 5 percent was considered significant. The reliability of the data was examined with Cronbach's coefficient alpha. Coefficients of determination for health promoting behavior were 42% in men and 48% in women. In both men and women, age, social ability of daily living and attitudes toward health-promoting behavior were related to health-promoting activity. In women, more actively going out was related to more active health promoting activity. Higher perception of transportation problems had a negative effect on going out. In men, neither of these factors had any relationship with health-promoting activity. In men, poorer health conditions were related to more active health-promoting activity, but in women, there was no relationship between those factors. These results show that there are gender differences in the relationships among the factors related to health-promoting activities in elderly people. In women, a higher perception of transportation problems restrained actively going out and health-promoting activity. PMID- 10388329 TI - [Life satisfaction and help needs in post-stroke patients]. AB - The purpose of this study was to investigate the life satisfaction and the help needs in post-stroke patients. A totaled 109 post-stroke patients were discharged from the rehabilitation ward of the Dokkyo University Hospital during two years from April 1995 to March 1997. The postal questionnaire was sent to 104 patients of them. The questionnaire was composed of two parts, one for the patients and one for their family members. The patients were asked about perceived improvement after discharge, outdoor activities, locomotor activities, and life satisfaction with their present state. Life satisfaction was assessed by using a visual analogue scale (VAS). The family members were asked about objective improvement of the patients after their discharge and their help needs. The distribution of the patients' life satisfaction showed a peak at around 50% by the VAS. While the perceived improvement and life satisfaction showed a significantly positive correlation, 9 patients (15%) recognized some improvement but marked their life satisfaction less than 50%. In help needs assessment, most family members classified them at a level of situational dependence or more. Only three cases were classified as dependence on individuals other than their family such as home help, which may suggest lack of social resources in their community. Logistic regression analysis revealed perceived improvement, going outdoors and age as significant adherent factors of life satisfaction, and objective improvement and age as those of help needs. Statistical analysis revealed a close association between perceived improvement and life satisfaction which have been suggested useful for QOL evaluation, but they must be interpreted as independent indicators. PMID- 10388330 TI - [The radar chart method and its analysis as a comprehensive geriatric assessment system for elderly disabled patients]. AB - In order to simply express the results of comprehensive geriatric assessment (CGA) for elderly disabled patients, we tried to develop a CGA system using a radar chart method in 50 patients (age 73-101, mean 85 +/- 5.4) admitted to our hospital during May 1997. Our clinical database for CGA included 7 major factors (diagnosis, mental function, physical function, nutritional state, complication, coronary risk factors, social background). Finally, the radar chart was made from the results of 6 scored factors other than diagnosis and the correlation was examined statistically between these factors. This study suggests that: (1) the radar chart method display of CGA is useful for all medical staff to understand the results of CGA for elderly disabled patients and the characteristic patterns of each disease, (2) because significant positive correlations were found between 3 factors (mental, physical, nutritional) in patients with cerebrovascular disease (CVD), a more global strategy for medical care planning, especially for treatment, nursing care and rehabilitation program is necessary in patients with CVD, (3) in patients with Alzheimer's disease (AD), significant positive correlation was found only between physical and nutritional factors; mental factors showed significant negative correlation only with the duration of morbidity and as a result, quality of life is a more important problem for planning care of patients with AD, (4) for elderly disabled patients, nutritional assessment and nutritional care planning are very important as well as mental and physical care planning. PMID- 10388331 TI - [Levofloxacin-induced neurological adverse effects such as convulsion, involuntary movement (tremor, myoclonus and chorea like), visual hallucination in two elderly patients]. AB - Levofloxacin-induced-neurological adverse events such as convulsion, involuntary movement (tremor, myoclonus and chorea-like) and visual hallucination in two elderly patients are reported. A 67-year-old man with minor alcoholism and a past history of gastrectomy and cholecystectomy was given 300 mg/day of oral levofloxacin and fulfenamic acid for an upper respiratory infection. On the 4th day, he reported gradual exacerbation of hand tremor which resembled chorea-like involuntary movement and gait disturbance. He also experienced visual hallucinations. On the 7th day, he suffered generalized convulsions and was admitted. Serum concentration of levofloxacin at this time (3 hours after last administration of a 100 mg tablet of levofloxacin) was 3.6 micrograms/ml. Cessation of the agents promoted complete recovery of these neurological adverse effects within a week. Another 85-year-old man with chronic bronchitis and slight renal impairment received long term administration of 200 mg/day of levofloxacin. On the 68th day of administration, gradual exacerbation of gait disturbance, dysarthria and chorea-like involuntary movement occurred. On the day of admission, 76 days after the start of administration, the serum level of levofloxacin was 2.55 micrograms/ml and that of spinal fluid was 1.12 micrograms/ml (3 hours after the last administration of a 100 mg tablet of levofloxacin). Cessation of the agents promoted complete recovery of these neurological adverse effects within the next two weeks. Both patients had no apparent neurological disorders except age-related brain atrophy. Age-related renal and brain impairment might have contributed to the neurological adverse effects of levofloxacin. PMID- 10388333 TI - Improving client outcomes through differentiated practice: a rural nursing center model. AB - The TriCounty Community Health Center (the Center) was created in 1994 with federal grant monies to increase access and to provide outreach and primary health care services for rural residents. The Center employs a differentiated practice model of nursing care in which all nurses use the nursing process targeted to client systems that match the nurse's level of educational preparation and competence. The model allows nurses to intervene with various client systems, including the individual, family, aggregate, and community. Program outcomes for the Center suggest that using a differentiated nursing practice model for outreach and primary care services appears to have a positive impact on the health of individuals, families, and aggregates in rural settings, using the Omaha Classification System as a framework for evaluation. PMID- 10388332 TI - A cholesterol intervention program for public health nurses in the rural southeast: description of the intervention, study design, and baseline results. AB - Residents of the rural South are at high risk for heart disease and are frequently identified as having high blood cholesterol, but sources for nutrition counseling in rural areas are often limited. To increase the availability of high quality nutrition counseling, the Food for Heart Program was developed for public health nurses and is designed to circumvent many of the obstacles common to dietary counseling. We conducted a randomized trial to assess the effectiveness of this program to lower blood cholesterol. In this report, we describe the study design, intervention program, and baseline characteristics of participants. Nurses at 17 health departments screened 781 subjects to enroll 468 with high blood cholesterol: three-quarters of the subjects were female, the mean age was 55, and 80% were white. Participants were at high risk for heart disease: 60% had two or more risk factors for coronary disease, the majority were overweight with a mean BMI of 29, and the mean cholesterol was 257 mg/dL. Reported baseline dietary intake included relatively modest consumption of high fat meats and snack foods, excessive consumption of sweets, modest intake of complex carbohydrates, and inadequate consumption of fruits and vegetables. PMID- 10388334 TI - Residential status and birth outcomes: is the rural/urban distinction adequate? AB - In studies comparing the birth outcomes of rural and urban women, residency status is frequently defined dichotomously as either rural or urban. Since residency status appears to be a continuum, however, the usefulness of other categorization systems needs to be explored. The purpose of this study was to compare birth outcomes using a three-level variable for residency status (rural, rural adjacent to urban, and urban). The study population was comprised of women who delivered by cesarean section over an 18-month period (N = 263) at a tertiary care hospital. Data were collected from patient charts, interviews, and the hospital information system. Residency status was determined by county of residence. Birth outcomes examined included gestational age, birthweight, Apgar scores, maternal complications, length of hospital stay, and costs of hospital care. Rural women had worse birth outcomes overall and traveled the greatest distance for delivery. Rural-adjacent women had the best birth outcomes of the three groups, yet were the youngest, least educated, least likely to be married, and the least likely to be privately insured. By using a nondichotomous three level variable for residency status, two distinct groups of rural women were identified whose maternal health care needs may differ from each other. PMID- 10388335 TI - The adolescent parenting program: improving outcomes through mentorship. AB - Adolescent parents and their infants are a population at risk. Infant mortality, low-birthweight, and child maltreatment are inordinately higher within this population than within slightly older cohorts. The purpose of this one group pretest-posttest intervention study was to analyze the efficacy of a program designed to improve infant outcomes through the enhancement of health practices and parenting skills in a sample of 137 low-income, pregnant and parenting adolescents who reside in an urban area and who screened positive for risk of child maltreatment. Based on theories of mentorship and social support, the program provided intensive home visitation by nursing paraprofessionals, indigenous to the community, for the 2 year study period. Program outcomes were compared to local and national data. Findings revealed only 4.6% of program infants were low-birthweight compared to local and national percentages of 13.5% and 9.42%. The mean length of gestation was 39.27 weeks (SD = 1.55). The incidence of infant mortality was zero, comparing favorably with national data as well as the local infant mortality rate (almost twice the state average). There were only four cases of child neglect, representing only 2.91% of the sample. This finding also compares favorably with national data. PMID- 10388336 TI - Conducting focus groups cross-culturally: experiences with Pacific northwest Indian people. AB - Many disciplines have used focus groups in research and the use has increased in the past 15 years (Smith, 1995). Procedural concerns have been explored, such as the selection of the participants, the location, and the size of the group, but little attention has been given to the consideration of cultural influences. The purpose of this paper is to focus attention on the impact of culture in conducting focus groups. Experiences from 15 focus groups conducted in two qualitative research studies with two Washington state Indian tribes over a 5 year period are presented and illustrate the importance of culture in conducting focus groups. Communication patterns, roles, relationships, and traditions were found to be important elements that must be considered in conducting focus groups cross-culturally. While some strategies discovered were found to be helpful, additional research is needed. PMID- 10388337 TI - Alternative measures of resource consumption in home care episodes. AB - Expected changes in home health care reimbursement will require a shift in focus from a visit-based unit to some other yet-to-be-defined unit of resource consumption. Little research has been done to understand other measures of resource consumption, however, especially those examining disciplinary differences. The purpose of this study was to provide empirical evidence on other measures of resource consumption as a way to frame discussions on alternative measures. Information is presented from a study of 102 home health care patients from 10 agencies in Ohio who completed an episode of care and remained at home. While the mean time per visit was similar for all disciplines (46 to 55 minutes), there were differences in the number of visits provided by various disciplines (home care aide services had the highest mean number of visits with 11.8). The mean cost per day for all services was $43.80 while the mean cost per episode was $1,160. Recommendations for further research include similar examinations using a more rigorous sampling methodology and including disparate populations of patients. PMID- 10388338 TI - A model for building collective capacity in community-based programs: the Elderly in Need Project. AB - As the focus of health promotion moves from individuals to organizations, communities and broader social policy, the models that guide public health program planning and development need reexamination. Public health nurses are gaining experience in strengthening and supporting the ability of communities to grow and change. This study aimed to illuminate the process. Data, gathered as part of an action research project to develop individual and community-based strategies to decrease isolation in frail older adults, provided a rich source of qualitative data. Analysis was directed toward identifying the factors and processes of capacity-building. The emerging model was clarified and partially validated with academics and practitioners from health promotion programs across the age span. Four stages of building collective capacity were identified: identifying common ground, working cooperatively, working in partnership, and working across the community. At each stage, processes of relationship building, project management and capacity-building resulted in stage specific products. A model of building collective capacity, grounded in community health practice and supported by the literature provides a base for developing practice indicators, and practice guidelines which will strengthen the ability to reach health goals. PMID- 10388339 TI - Articulating the culture and tradition of community health nursing. AB - While community health nursing (CHN) leaders speculate about the future, nurses on the front lines care for vulnerable families and populations in the midst of diminishing resources, radical changes in health care delivery systems, and unwieldy bureaucracies. Narrative data from a recent interpretive study provided an unexpected opportunity to explore how CHN practice in diverse settings is evolving in response to such changes. Data consisted of interviews and observations of 25 nurses in their practice setting. Several clinical stories or exemplars are selected to highlight how the "culture" of agency settings shapes public health nursing (PHN) practice in ways that need to be recognized and strengthened or affirmed. Clinical storytelling can play a crucial role in preserving the PHN tradition and restoring and transforming local cultures when PHNs, administrators, educators, and researchers commit to PHN excellence. PMID- 10388340 TI - Use of the SF-36 to identify community dwelling rural elderly at risk for hospitalization. AB - The SF-36 has been identified as a generic measure of health-related quality of life outcomes which is not age, disease, or treatment specific and is deemed appropriate for monitoring the results of care as well as a measure of outcomes from the patient's perspective. The purpose of this study was to use the SF-36 to assess the general health status and health promotion activity of rural elderly 1 year following participation in a community-based health promotion project. Eighty participants from the original sample of 222 (190 community-based and 32 home-based) were contacted by telephone and the SF-36 was administered. Results found that 76% of the study participants were correctly predicted for past hospitalization. While this project studied elderly in a rural community, the administration of this instrument can be useful in identifying a variety of populations at risk for hospitalization. PMID- 10388341 TI - Expanding urban learning experiences for non-traditional students. AB - Educating nurses in the motivation and ability to provide appropriate and quality health care to urban inner-city residents with complex and multiple health problems has been a continuing challenge to academic institutions. Recruiting appropriate students and providing meaningful learning experiences is the first of many challenges. Understanding and addressing the many barriers to accessing health services is an important learning outcome. Successful providers with underserved populations have been found to have a strong sense of service to humanity and pride in making a difference and have thrived on the challenges of creatively using limited resources to deal with their patients' complex needs. Establishing a Returned Peace Corps Fellows program and a community health nursing track within the undergraduate program in nursing has provided some successes and additional answers. While studying for a professional degree, the Fellows are placed in a service position to integrate their Peace Corps experiences into new professional learning as it is taking place and to earn a stipend to assist with the cost of their education. This has led to the development of a community health nursing track in the undergraduate program, a combination of required-for credit courses, credit-earning enrichment and independent study experiences, and stipend-earning clinical experiences outside the curriculum. PMID- 10388342 TI - Collective conscience: the ethics committee and community. PMID- 10388343 TI - Inborn defects of fatty acid oxidation: a preventable cause of SIDS. AB - Inborn errors of fatty acid oxidation, including medium chain acyl CoA dehydrogenase (MCAD) deficiency are readily detectable and treatable metabolic disorders in which recognition of symptoms is important. Symptoms occur when there is fasting, often associated with illness. If not diagnosed, these inborn errors of metabolism can result in sudden death classified as SIDS. These disorders can be diagnosed by ordering plasma or blood spot acylcarnitine profiles. PMID- 10388344 TI - Chronic acalculous cholecystitis: reproduction of pain with cholecystokinin and relief of symptoms with cholecystectomy. AB - Over 500,000 patients undergo cholecystectomy annually in the United States for symptoms of upper abdominal discomfort and pain ascribed to gallbladder disease. However, approximately 5%, or 25,000 of these cases do not have gallstones on ultrasound examination but typically present with chronic symptoms of biliary colic. These patients often present as challenging diagnostic dilemmas and are often treated as if their symptoms are secondary to peptic ulcer disease or other gastrointestinal-related disorders. In 1992, we began to use the cholecystokinin (CCK) challenge test on patients with normal ultrasound examinations of the gallbladder but who had chronic symptoms resembling biliary colic. The CCK test was considered positive if the identical symptoms of discomfort or pain, usually in the right upper quadrant of the abdomen, were reproduced. This study describes the first 24 patients who had a positive CCK challenge test and chose to undergo cholecystectomy for relief of their symptoms. No patient was lost to follow-up evaluation at 1 to 24 months after operation. PMID- 10388345 TI - Selective serotonin reuptake inhibitors (SSRIs) and falls in the elderly depressed patient. PMID- 10388346 TI - [Prevention of tuberculosis in Denmark]. PMID- 10388347 TI - [The global epidemiology of tuberculosis]. PMID- 10388348 TI - [Early diagnosis of tuberculosis. Proposed paradigmatic shift]. PMID- 10388349 TI - [Tuberculosis research. How far have we advanced and where does it lead?]. PMID- 10388351 TI - [Tuberculous meningitis in children]. AB - Medical records of six children with tuberculous meningitis were reviewed. The patients were admitted to the paediatric departments in the city and country of Copenhagen between 1983 and 1997. Interpretation of cerebrospinal fluid findings, and difficulties in early recognition of tuberculous meningitis are discussed. PMID- 10388350 TI - [Tuberculosis and migration]. AB - The decline of tuberculosis in many western countries has faded off in many countries in recent years because of migration of people from high-prevalence to low-prevalence areas. Tuberculosis in immigrants mirrors tuberculosis in the country of origin with a predominance of young people with a high incidence of extrapulmonary tuberculosis. Typically, the tuberculosis appears in the first five years after arrival. The impact of immigrant tuberculosis has been discussed in a number of recent publications. In Denmark the incidence of tuberculosis in immigrants now outnumbers the incidence in Danes. Screening for tuberculosis in immigrants is not obligatory and is only offered to asylum seekers under the care of the Danish Red Cross but could be considered in all immigrants from high prevalence countries. PMID- 10388352 TI - [Occurrence of tuberculosis among children in the area of Copenhagen 1984-1993]. AB - This study was undertaken to describe the epidemiology, clinical manifestations and prognosis of childhood tuberculosis in Copenhagen, with special attention to differences between Danish children and children of foreign origin. Medical records for all children with tuberculosis cared for in the hospitals of the Copenhagen area 1984-1993 were reviewed. Sixty-six patients were identified. Sixteen of 20 Danish patients (80%) and 67% of foreign children had respiratory tuberculosis. Tuberculosis located in cervical lymph nodes was found only in children of foreign origin. Five patients had meningitis. The high incidence among foreign children reflects the incidence in their home countries, but poorer living conditions among ethnic minorities in Denmark may facilitate transmission of tuberculosis. Severe manifestations of tuberculosis still occur, even in a low incidence country. PMID- 10388353 TI - [Tuberculosis in Denmark 1972-1996]. AB - The present study is based on notified cases of tuberculosis (TB) in the National tbc. register 1972-1996. A decline in Tb incidence was seen from 1972 and until the mid-1980's. Subsequently the trend has reversed due to an increasing number of TB cases in foreigners. In 1996, 60% of all cases of TB in Denmark were found in foreigners reflecting the rising number of refugees and their families arriving in Denmark from highly endemic areas, mainly Somalia. Among native Danes the TB incidence fell from 14 per 100,000 in 1972 to 4 per 100,000 in the 1980's and stabilized at this very low level. The unchanged incidence in Danes covers a falling incidence in the older and a rising incidence in the younger and middle aged adult population, mainly in the capital. Approximately half of the cases occur in high-risk groups. The TB-epidemic is close to elimination in the indigenous Danish population, but the disease is maintained at a low level probably due to increased patient and doctor delay and resulting microepidemics primarily in high-risk populations. PMID- 10388354 TI - [An outbreak of Pontiac fever among children and adults following a whirlpool bath]. AB - We investigated an outbreak of fever most likely due to a contaminated whirlpool among nine adults and six children visiting a holiday home. The outbreak was characterized by a high attack rate, short incubation periods, influenza-like symptoms and rapid recoveries typical of Pontiac fever. The children, however, experienced less characteristic symptoms and no sequelae compared to the adults. Evidence and presumptive evidence of Legionella (L) infection was found in eleven cases; in one case by isolation of L. pneumophila serogroup 1, in two cases by positive test for Legionella by PCR and in eleven cases with seroconversion. In contrast, two adult non-users of the whirlpool had no symptoms and no serological evidence of infection. This investigation demonstrates differences between adults and children in the clinical picture of Pontiac fever, furthermore it shows that culture and PCR of tracheal aspirate for legionellae can be used in a hospital setting for rapid diagnosis although their sensitivities are low. PMID- 10388355 TI - [Hereditary neuropathy with liability to pressure palsies]. AB - Hereditary neuropathy with liability to pressure palsies (HNPP) is an autosomal dominant disorder characterized by recurrent transient pressure palsies of peripheral nerves and slowing of nerve conduction velocity of the peripheral nerves at common sites of compression. In most cases the molecular basis of the disease is a 1.5 Mb deletion on chromosome 17p11.2. We report four members of a family with different clinical phenotypes. Electrophysiological and genetic studies were consistent with the diagnosis of HNPP. Nerve biopsy is only necessary in patients with a normal result of the molecular genetic analysis. The variability of the clinical phenotype along with asymptomatic individuals could account for an under-recognition of this inherited neuropathy. PMID- 10388356 TI - [Picture of the month. Pulmonary tuberculosis]. PMID- 10388357 TI - [Pet animals and allergy]. PMID- 10388358 TI - [Meeting around a blackboard]. PMID- 10388359 TI - [Environment and cancer. Necessary prevention of cancer]. PMID- 10388360 TI - [Is radical prostatectomy to be introduced?]. PMID- 10388361 TI - [Barefoot treatment of depression?]. PMID- 10388362 TI - [Criticism of the treatment of varicose veins should have a valid basis]. PMID- 10388363 TI - [On the treatment of chronically incapacitated]. PMID- 10388364 TI - [Cerebral or mental diseases]. PMID- 10388365 TI - [Acupuncture therapy of back pain seen from a perspective of MTV]. PMID- 10388366 TI - [Myoses]. PMID- 10388367 TI - Effect of topical fluoride application before and after amalgam restoration placement on recurrent caries inhibition. AB - PURPOSE: To evaluate the in vitro caries inhibition effects of 1.23% APF foam topical fluoride treatment of cavity preparations, prior to restoration placement and after restoration placement. MATERIALS AND METHODS: Sixty standardized Class V preparations were placed in molars. Randomly, 40 teeth received an amalgam restoration: Twenty other teeth had 1cc 1.23% APF foam applied to preparation surfaces for 1 minute, then an amalgam restoration was placed. The APF was not rinsed away prior to restoration, it was displaced by the pressure of the amalgam being condensed into the preparation. Twenty of the initial 40 amalgam restorations had 1 cc 1.23% APF foam applied to the external tooth/restoration margins for 4 minutes, the remaining 20 amalgam restorations acted as the controls. Acid-resistant varnish was placed, leaving 1 mm of tooth adjacent to restoration margins exposed. All specimens were subjected to an artificial caries challenge (pH 4.4) for 5 days. Sections of 100 microns were cut longitudinally through the restored margins, photographed under polarized light microscopy, then demineralized areas adjacent to restoration margins were quantitated. RESULTS: Results demonstrated the mean (+/- S.D.) area (microns 2) of demineralization 100 microns from the restoration margins to be: amalgam 10,221 +/- 524, amalgam with 1-minute APF over preparation 529 +/- 557, amalgam with 4-minute APF over external surface 736 +/- 359. ANOVA and Duncan's (P < 0.05) indicated both 1.23% APF foam topical fluoride treatment regimens exhibited significantly less demineralization at restoration margins than the non-treated amalgam control. There was no significant difference between the two fluoride placement regimens examined in this study. PMID- 10388368 TI - Bonding to dentin. Clues to the mechanism of adhesion. AB - PURPOSE: To investigate the effect on shear bond strength of the presence or absence of the demineralized collagen layer on both wet and dried etched dentin surfaces. MATERIALS AND METHODS: Composite cylinders were mounted on dentin surfaces after the following treatments; 37% etch/wet, 37% etch/dry, 37% etch/NaOCl/wet, 37% etch/NaOCl/dry, No-etch/wet, No-etch/dry, No-etch/NaOCl/wet, No-etch/NaOCl/dry. 24-hr shear bond strength data was obtained. SEM examinations of various surfaces were also conducted. RESULTS: Wet bonded specimens had significantly higher shear bond strengths, with or without the presence of the demineralized collagen layer, whether they were acid-treated or not. Acid-etched specimens had significantly higher shear bond strengths than unetched specimens. PMID- 10388369 TI - Microleakage of Class II box-type composite restorations. AB - PURPOSE: To investigate whether in small box-type Class II preparations the use of glass ionomer cement, a dentin primer or a different type of conditioner had an effect on the microleakage compared to a more simple total-etch system using a phosphonated bonding agent. MATERIALS AND METHODS: In extracted premolars, 84 box type Class II composite resin restorations with margins in enamel were made following a standardized protocol. A transparent matrix system was used and the composite resin was applied in two layers. The teeth were restored using a phosphonated total-etch bonding system as a control (Photo Bond). In the experimental groups a glass ionomer lining-cement (Vitrebond), a total-etch adhesive including a primer (Scotchbond Multi-Purpose) and a total-etch adhesive using a self-etching primer (Liner Bond 2) were used. The teeth were thermocycled, immersed in a dye solution and sectioned. The cervical and occlusal dye penetration was assessed. RESULTS: Preventing microleakage was not more effective in the experimental groups than in the control group (P < 0.05). Scotchbond MP groups using maleic acid showed statistically significant more microleakage than the control group (Photo Bond) at the cervical side. Liner Bond 2 and Scotchbond MP groups using maleic acid showed statistically significant more microleakage than the control group at the occlusal side. The use of a lining cement did not improve the quality of the restoration. From this study it was concluded that in this type of restoration, total-etch systems using a primer are not always an improvement. PMID- 10388370 TI - Directed polymerization shrinkage versus a horizontal incremental filling technique: interfacial adaptation in vivo in Class II cavities. AB - PURPOSE: (1) To evaluate the interfacial adaptation to dentin and enamel of Class II composite resin restorations placed in vivo with the directed shrinkage technique, a combination of a self-curing (Bisfil 2B) and a light-curing composite resin (AElitefil); (2) To compare this technique with a horizontal incremental filling technique, where the gingival layer of the light-curing composite resin was cured with a transparent light-tip; (3) To evaluate the effect of a hydrophilic bonding system (All-Bond 2) on the marginal adaptation of both application techniques. MATERIALS AND METHODS: In each of 34 premolars, scheduled to be extracted for orthodontic reasons, a mesial and a distal cavity was restored with composite resin filling material using the directed shrinkage technique (Bisfil 2B/AElitefil) and a horizontal incremental filling technique (AElitefil). In six groups, a hydrophilic dentin bonding system (All-Bond 2) was applied. As control, an enamel bonding agent (Gluma 4) was used in one of the directed shrinkage and in one of the horizontal incremental filling groups. For conditioning of the cavities a 10% phosphoric acid gel was used in six groups and a 32% acid gel in the other two groups. The teeth were extracted after 1 month, sectioned and replicas of the sections were made. Quality of the interfacial adaptation was studied with a scanning electron microscope. RESULTS: On the pairwise comparisons between the two application groups, no significant differences were found between the directed shrinkage technique and the horizontal incremental filling technique. The groups using the hydrophilic bonding system showed a significant better adaptation, with gap-free attachment in 77%-87% of the length of the dentin margins investigated. No significant differences for adaptation to enamel were seen between the investigated groups. The adhesive failures were found mostly between the hybrid layer and the composite resin, while the dentin was still sealed. No significant difference in interfacial quality was seen between the cavities using the 10% or the 32% phosphoric acid conditioning. A relative high frequency of enamel fractures, parallel to the interfaces, was found in the 10% phosphoric acid-conditioned cavities, especially in the cervical enamel interfaces. PMID- 10388371 TI - Soluble calcium/SMFP dentifrice: effect on enamel fluoride uptake and remineralization. AB - PURPOSE: To evaluate in situ the effect of soluble calcium on fluoride uptake and remineralization by human dental enamel from a sodium monofluorophosphate (SMFP) dentifrice. MATERIALS AND METHODS: Eight volunteers took part in this cross-over, double-blind design study performed in three phases of 45 days. During each phase of the experiment, the subjects wore an acrylic resin appliance containing four blocks of human dental enamel with artificial caries to evaluate fluoride uptake and remineralization by three dentifrices: SMFP containing CaCl2, SMFP, and placebo. After each phase, the enamel blocks were removed and the total fluoride uptake (ppm F) and microhardness (Knoop) were measured. Statistical analyses (ANOVA and Turkey's test) were done. RESULTS: There was no statistically significant difference between SMFP-CaCl2 and SMFP treatments regarding the amount of fluoride and microhardness data of treated enamel blocks as well as their areas under the curves, although both differed significantly (P < 0.05) when they were compared with placebo. However, the SMFP-CaCl2 treatment demonstrated the highest values on fluoride uptake and microhardness data. Furthermore, the microhardness data demonstrated 50% and 40% of remineralization for SMFP-CaCl2 and SMFP dentifrices, respectively. PMID- 10388372 TI - Short-term clinical evaluation of post-operative sensitivity with bonded amalgams. AB - PURPOSE: To compare the in vivo short-term post-operative sensitivity of teeth restored with amalgam using a bonded resin liner vs. teeth restored using a copal varnish liner. MATERIALS AND METHODS: 20 patients received Class I or Class II contralaterally paired restorations which were placed at the same appointment. All restorations were placed by the same operator using an identical technique except that, in each randomized pair, one was lined with an adhesive resin (Scotchbond Multi-Purpose Plus) while the other was lined with copal varnish. (Plastodent) Patients were provided visual analog scale response forms, instructed in their use, and requested to complete and return a form reporting their degree of sensitivity at baseline and on days 1, 3, 7, 14, and 30 post operatively. Data from the response forms were analyzed for differences using a paired t-test. RESULTS: A response rate of 90% (18/20) was achieved for the complete 30-day assessment. Increases in thermal sensitivity beyond baseline were seen in 13 of the 18 subjects involving 12 restorations lined with copal varnish and 10 lined with adhesive resin. Typically, sensitivity peaked on day 1 or day 3 and diminished to pre-operative levels by day 30. Only three subjects reported greater sensitivity at day 30 than at baseline. No significant difference in post operative sensitivity was found between the two cavity lining materials at any post-operative interval. PMID- 10388373 TI - Fracture strength of weakened human premolars restored with amalgam with and without cusp coverage. AB - PURPOSE: To determine the effect of cusp coverage with amalgam restorations on the fracture strength of weakened human maxillary premolars. MATERIALS AND METHODS: 30 extracted human maxillary premolars were divided into three groups; Group A (control): uncut teeth, Group B: wide MOD cavities prepared and the pulp chamber's roof removed. The teeth were restored with amalgam without cusp coverage, Group C: same as Group B plus cusp reduction and restored with amalgam. The teeth were included in metal rings with self-curing polystyrene resin and stored in water for 24 hrs. The restorations were refined with rubber points, polished and further stored in water for 24 hrs before being subjected to a compressive axial load in a Universal testing machine at 0.5 mm/min. RESULTS: The mean fracture strength obtained was: Group A: 173.3 Kgf, Group B: 47.3 Kgf and Group C: 127.7 Kgf. All values were statistically significantly different (P < 0.05). The amalgam restoration of weakened human premolars with cusp coverage amalgam significantly increased the fracture strength of the teeth (63%) as compared to teeth restored without cusp coverage. The results showed that cusp coverage with amalgam might be an option for restoring weakened endodontically treated teeth. PMID- 10388374 TI - Effect of ultrasonic cleaning on microorganisms. AB - PURPOSE: To establish a method to measure microbial kill caused by ultrasonic cleaning. Secondarily, to estimate the escape of bacteria from the ultrasonic cleaning solutions during operation of the unit. MATERIALS AND METHODS: Three commercial enzymatic detergents and saline were used as cleaners. Depending on detergent, initial operational temperature was 21 degrees C, 37 degrees C or 60 degrees C. Streptococcus mutans ATCC 25175 (S. mutans suspensions) was adjusted to a final concentration of 1.0 x 10(3) cells/mL in saline. Suspensions (2000 mL) at the desired temperatures were added to the cleaner. Aliquots were removed, serially diluted in letheen broth and spread plated over mitis salivarius agar. Appropriate amounts of detergent solutions were added to S. mutans suspensions and the cleaner operated for 20 minutes. Aliquots were then removed and plated. The process was repeated twice. Plates were aerobically incubated at 37 degrees C for 7 days and the colonies counted. The procedure was repeated using three temperatures of S. mutans suspensions (21 degrees C, 37 degrees C or 60 degrees C), but without detergent or ultrasound. Also, detergents were added to 21 degrees C S. mutans suspensions and allowed to sit for 20 minutes without ultrasonic cleaning. Microbial sampling was done as previously described. RESULTS: Results when ultrasound was used indicated that little kill (5-15%) occurred in 21 degrees C or 37 degrees C detergent solutions. Greater kill (25 35%) was noted with 21 degrees C and 37 degrees C saline. Complete kill was accomplished with 60 degrees C saline or the 60 degrees C detergent solution. When ultrasound and detergent were not used, there was no kill in 21 degrees C and 37 degrees C saline, but complete kill in 60 degrees C saline. In the absence of ultrasound no kill was noted in 21 degrees C S. mutans suspensions to which detergent had been added. Total kill of S. mutans was observed in 60 degrees C saline or 60 degrees C detergent with ultrasound or after a 20-minute exposure in 60 degrees C saline without ultrasonic cleaning. Very few bacteria escaped from the ultrasonic cleaning solutions into the air during the cleaning process. Placement of the unit lid effectively reduced emissions to zero. PMID- 10388375 TI - Extension for prevention: is it relevant today? AB - Extension for prevention has been an integral part of dentistry for over 100 years. Because this concept advocated the removal of sound tooth structure, it was not totally accepted at the turn of the century. The advent of the gold casting catapulted extension for prevention into general acceptance. In 1883, Webb presented a concept of "prevention of extension of decay". This concept advocated a proximal cavity preparation extending toward the buccal and lingual aspects of the tooth so that contact with adjacent teeth would not be at the margins. The separation of the margins, along with proper restoration contours, was thought to promote natural cleansing of the embrasures with saliva and fluids in the diet. GV Black's 1891 idea of "extension for prevention" was to provide extension of the preparation to the facial and lingual line angles in order to bring about "self-cleansing" margins via food excursion. Black's concept also included extending preparations through fissures to allow cavosurface margins to be on non-fissured enamel. Black integrated the extension of the proximal margins with his concept of an occlusal isthmus for a Class II amalgam preparation one third the faciolingual width of the occlusal surface. Challenges to this concept of extension for prevention were immediate; and, by the 1950's, narrower, more conservative preparations were seen by a few as being more effective in preserving teeth. Not only occlusal width was reassessed, but the need to routinely extend proximal margins to the buccal and lingual line angles was also questioned. By the mid-1960's and early 1970's a more conservative approach to amalgam preparation was advocated and was being taught in some dental schools. Today, a standardized outline form should not be used or taught as a principle of cavity preparation. In areas where fissure caries has necessitated a preparation extending into dentin, a composite resin or dental amalgam restoration should be placed, and a fissure sealant should be used to protect remaining susceptible fissures from carious attack. This current form of the concept of extension for prevention, which is supported by clinical research, preserves sound tooth structure that, using outdated concepts, would have been cut away. Placing proximal margins in sound tooth structure that just clears an adjacent tooth is also strongly advocated. Sound enamel margins in certain areas may occasionally be left in contact with adjacent teeth for amalgam preparations. For Class II preparations for composite resin, facial or lingual proximal bevels will usually suffice to separate the margins from the adjacent tooth to allow finishing and polishing at the margins. Preventing unnecessary extension and allowing sounder tooth structure to remain is one important aspect of helping patients to maintain their teeth for their lifetimes. PMID- 10388376 TI - Designs for cast metal restorations. AB - This article reviewed clinical conditions that assist selection of specific design decisions for the tooth preparation for cast metal restorations. This decision should result in a conservative restoration for a given clinical situation. A paradigm is included to assist the dentist in the decision-making process. PMID- 10388377 TI - Influence of different etching times on hybrid layer formation and tensile bond strength. AB - PURPOSE: To correlate the thickness of the dentin/resin hybrid layer and the influence of different etching times to the composite resin tensile bond strength. MATERIALS AND METHODS: 775 human molars were prepared by removing the occlusal cusps to expose middle dentin using a microtome saw. Five commercially available dentin bonding agents were applied according to the manufacturers' instructions. Only the etching times were varied. For confocal laser scanning microscopy (CLSM) measurements, 325 teeth were used and the etching times were varied (not etched, 15, 30, 60 or 120 seconds). The primer components or self priming adhesive, respectively, were fluorescence-labeled by rhodamine B isothiocyanate. In each case, the prepared surfaces were then covered by a layer of composite resin, sectioned and examined using a CLSM in fluorescent mode. The extension of hybrid layers was quantified by measuring the distances between the dentin-composite junction to a visible boundary. For the tensile bond strength measurements (Zwick testing machine), 450 molars were prepared, using different dentin etching times (not etched, 15, 30, 60, 120 and 180 seconds). RESULTS: A hybrid layer was found in all etched teeth. The CLSM data gave evidence for an exponent-function relationship between the periods of etching and the thickness of the hybrid layer. The highest tensile bond strengths were achieved after 15 s of etching, followed by 30 s and 60 s. Under these etching conditions and irrespective of the bonding agent, bond strengths were significantly higher (P < or = 0.05) than without etching or after 120 and 180 s of etching. There was no linear correlation between the thickness of the hybrid layer and the bond strength. PMID- 10388378 TI - Effect of a re-wetting agent on the performance of acetone-based dentin adhesives. AB - PURPOSE: To compare the in vitro bond strengths of two acetone-based one-bottle dentin adhesives applied to four surface moisture conditions. The tested hypothesis was that wetting a dried dentin surface with an aqueous HEMA solution would result in bond strengths similar or higher than those obtained by leaving the surface moist as per manufacturers' instructions. MATERIALS AND METHODS: Eighty flat dentin bonding sites were polished to 600-grit on middle dentin of the labial surface of bovine incisors mounted in acrylic resin. The specimens were equally and randomly divided between two acetone-based dentin adhesives (One Step and Prime & Bond 2.1) and four different levels of surface moisture (moist dentin, dentin dried for 1 s, dentin dried for 5 s, and dentin dried for 5 s followed by re-wetting with Aqua-Prep, an aqueous HEMA solution). A composite post was then adapted to the treated area and light-cured. After thermocycling, the bond strengths were determined by testing the specimens in shear. Field Emission SEM examinations were carried out to evaluate the effects of different treatments on the dentin-resin interface. RESULTS: Statistical analysis revealed that the application of One-Step resulted in similar mean shear bond strengths for the groups in which moisture was present on the dentin surface (12.0-14.2 MPa). The mean shear bond strengths for the group in which One-Step was applied to a dried dentin surface was significantly lower (6.0 MPa). For Prime & Bond 2.1, the application of a re-wetting solution significantly increased mean shear bond strengths (13.9 MPa). The remaining three Prime & Bond 2.1 groups yielded statistically similar mean bond strengths, regardless of the surface condition (6.6-8.1 MPa). PMID- 10388379 TI - Evaluation of cure, properties and wear resistance of Artglass dental composite. AB - PURPOSES: (1) To evaluate the degree of conversion (DC) and the physical properties of a new dental composite, Artglass (Kulzer), designed primarily as a replacement material for porcelain in PFMs; and (2) to compare the efficacy of two different curing units for two dissimilar composites (Artglass and Charisma, Kulzer). MATERIALS AND METHODS: Specimens (n = 10) were prepared by curing either in a continuous light exposure unit (Triad II, Dentsply) or in a high intensity strobe unit (Uni XS, Kulzer) for 180 s. Specimens were aged 24 hrs in water at 37 degrees C and tested for fracture toughness (FT-MPa m1/2; x-head = 0.13 mm/min), flexural modulus (E, GPa), flexural strength (FS, MPa; x-head = 0.5 mm/min), hardness (KHN, kg/mm2), and in vitro wear resistance (OHSU oral wear simulator). DC (%) was determined by transmission micro-FTIR. Results were compared by ANOVA/Turkey's test (P < or = 0.05). RESULTS: DC, FT, E and FS were improved for Artglass and DC and E were improved for Charisma when the strobe curing unit was used, probably due to its increased intensity. Artglass showed a greater DC and FT but a lower E, KHN and wear resistance compared to Charisma when similar curing methods were used. PMID- 10388380 TI - Effect of wet and dry bonding techniques on marginal leakage. AB - PURPOSE: To evaluate the effect of wet and dry bonding on microleakage of Class V restorations bonded with three bonding agents. MATERIALS AND METHODS: 60 extracted human premolars and molars were randomly assigned to three groups for bonding with Gluma CPS, EBS (ESPE) and Prime & Bond 2.1. Cavities were cut in both the buccal and lingual surfaces. Half of each preparation was in enamel and the other was in cementum/dentin. The cavities were restored with composite after the application of dentin bonding agents using a wet and dry technique for each material. The teeth were stored in distilled water for 6 days at 37 degrees C, thermocycled, and the restorations examined microscopically for leakage using Procion Brilliant Red as a marker. RESULTS: All groups showed microleakage at both the enamel and dentin margins. At the gingival margin, there was a significant difference between the groups for both wet-bonding (P = 0.039) and dry-bonding (P = 0.024). There was no significant difference between the groups at the enamel margin (wet bonding: P = 0.179, dry-bonding: P = 0.357). The wet bonding technique was compared with the dry-bonding technique for each material at both the gingival and enamel margins and no significant differences were observed (in all cases P > 0.47). SEM showed that in dye-labeled areas debonding occurred mostly, but not always, near the resin-hybrid interface. PMID- 10388381 TI - Effect of primer solvent, primer agitation, and dentin dryness on shear bond strength to dentin. AB - PURPOSE: To evaluate the shear bond strength of acetone- or water-based primers applied with or without agitation to either wet or dry dentin. MATERIALS AND METHODS: Forty-eight caries-free extracted human premolars were embedded in gypsum and flat dentin surfaces were prepared. The teeth were randomly divided in eight groups with six specimens each. After acid etching and rinsing, either a HEMA and acetone (Ac) or a HEMA and water (Wa) primer were applied to dry (D) or wet (W) dentin. The primer was placed with (Y) or without (N) agitation according to the following combinations: Group 1: D-Ac-Y; Group 2: D-Ac-N; Group 3: D-Wa-Y; Group 4: D-Wa-N; Group 5: W-Ac-Y; Group 6: W-Ac-N; Group 7: W-Wa-Y; Group 8: W-Wa N. The primed surface was dried, covered with a bonding resin and light-cured, whereupon a composite cylinder was bonded to the dentin surface. The shear bond strength was determined after 30 days in water. RESULTS: The mean and standard deviations, expressed in MPa, were: Group 1: 13.1 +/- 3.42; Group 2: 15.9 +/- 4.84; Group 3: 19.8 +/- 6.64; Group 4: 18.6 +/- 5.18; Group 5: 16.5 +/- 5.25; Group 6: 25.5 +/- 4.79; Group 7: 21.9 +/- 4.94; Group 8: 17.4 +/- 4.50. The results were analyzed using ANOVA and Waller-Duncans K-ratio t-test. The acetone based primer gave the highest bond strength to wet dentin without agitation. When the acetone-based primer was used on dry dentin a significant decrease in bond strength occurred (P < 0.05). No significant difference (P > 0.05) was found for the water-based primer on wet or dry dentin. Agitation improved the bond strength for the water-based primer, but decreased the bond strength for the acetone-based primer. PMID- 10388382 TI - Class I occlusal composite resin restorations: in vivo post-operative sensitivity, wall adaptation, and microleakage. AB - PURPOSE: To investigate the effect of restoration technique and adhesive system on the post-operative sensitivity and marginal adaptation of Class I occlusal composite resin restorations placed in vivo. MATERIALS AND METHODS: 48 Class I cavities were restored in vivo according to one of three protocols: (1) Scotchbond Multi-Purpose/P50 placed in increments; (2) Scotchbond Multi Purpose/P50 placed in bulk, and (3) Clearfil Liner Bond 2/Clearfil Ray Posterior placed in bulk. Post-operative sensitivity and sensitivity on loading were recorded 5-7 weeks after placement of the restorations; the teeth were cautiously extracted, immersed in a dye solution and sectioned. SEM observations were made from epoxy resin replicas. Microleakage and gap formation was assessed. RESULTS: No differences among adhesive systems or restoration procedures were found for microleakage. Post-operative sensitivity was reported in 14% of all teeth but was absent in the Clearfil Liner Bond 2 group. Sensitivity on loading was experienced by patients in 56% of the restorations. Group 1: nine teeth; Group 2: 15 teeth; Group 3: three teeth. Differences were statistically significant for all three groups. The SEM analysis showed that restorations placed in two layers showed less gaps than restorations placed in bulk. PMID- 10388383 TI - Bonding mechanism of Ketac-Molar Aplicap and Fuji IX GP to enamel and dentin. AB - PURPOSE: To evaluate the bonding mechanism to enamel and dentin of two chemically cured restorative glass ionomer cements (GICs). MATERIALS AND METHODS: Buccal surfaces of six caries-free premolars stored in a 1% chloramine solution were ground and then polished with 600 grit SiC paper to expose both enamel and dentin. Ketac-Molar Aplicap (after conditioned with Ketac-Conditioner) or Fuji IX GP (after conditioned with Cavity Conditioner) was applied on the enamel and dentin according to the manufacturers' instructions. Ketac-Molar Aplicap was coated with Ketac-Glaze and Fuji IX GP with GC Fuji Varnish. After 24-hr immersion in water, all teeth were sectioned bucco-lingually in the center of the bonded surface. Bonded interfaces of one of the halves of the sectioned specimens were etched with 35% phosphoric acid gel for 10 s and then soaked in 5% sodium hypochlorite solution for 5 min. All specimens were dehydrated with ethyl alcohol and dried with HMDS. Bonding interfaces were coated with gold-palladium and observed under the SEM. RESULTS: For both Ketac-Molar Aplicap and Fuji IX GP, an intimate adaptation between the material and the enamel was observed. Both materials bonded to dentin without gap formation. For both Ketac-Molar Aplicap and Fuji IX GP, neither resin tags nor hybrid layer was observed in any specimen. PMID- 10388384 TI - Effect of cement space and delayed placement on the seating of crowns luted with Vitremer, Fuji Duet and Dyract Cem. AB - PURPOSE: To determine the effects of cement space and delayed placement on the post-cementation elevation of crowns luted with two resin-modified glass ionomer luting cements (Vitremer, Fuji Duet) and a self-curing polyacid-modified composite resin luting cement (Dyract Cem). MATERIALS AND METHODS: The investigation was conducted in two parts: (1) A loading apparatus was employed to simulate the seating of a crown on a die. Six dies, differing in the amount of cement space allowed from 6-52 microns, were used. The loading apparatus allowed for a test cement to be placed in the crown, and subsequent activation of the apparatus lead to seating of the die into the crown. After seating, post cementation crown elevation was measured. The cements Phosphacap, Vitremer, Dyract Cem and Fuji Duet were used. Each cement was tested 10 times at each of the six cement spaces. The post-cementation crown elevation data was analyzed by one- and two-way ANOVA tests, and Tukey's HSD test. (2) The cements Vitremer, Fuji Duet and Dyract Cem, were tested in a similar fashion to Part 1, except activation of the loading apparatus was delayed. Three cement spaces (6, 29 and 52 microns) and three delay periods (60, 120 and 180 seconds) were investigated. Each cement was tested five times. The post-cementation crown elevation data was analyzed by one- and two-way ANOVA tests, and Tukey's HSD test. RESULTS: The amount of cement space provided and the cement used significantly affected post cementation crown elevation (P = 0.001). Where cementation was delayed, post cementation crown elevation was significantly affected by both the cement used and the amount of cement space provided (P = 0.001). Delayed placement of Dyract Cem had no significant effect on post-cementation crown elevation (P = 0.01). Post-cementation crown elevation when Vitremer was used was significantly affected by both the amount of cement space and delayed placement (P = 0.01). A 180-second delay in placement of Vitremer resulted in significantly worse post cementation crown elevation (P = 0.01). Seating was not possible 120 seconds after Fuji Duet had been mixed. PMID- 10388386 TI - Caries removal techniques and instrumentation: a review. AB - The invention of rotary instruments not only improved the speed of caries removal but also the destruction of sound tooth substance. Hence, as early as the 1950s, there were attempts to develop a less invasive technique, such as the air abrasive and ultrasonic technique, for the purpose of caries removal. The proposed use of air-polishing was published in the early 1980s. Subsequent better understanding of the carious process saw the introduction of the enzyme technique in the late 1980s. Other techniques, such as chemomechanical caries removal and laser systems, have also been attempted and researched during the last four decades to minimise the unnecessary removal of sound tooth substance, although these and other techniques reviewed in this article have not yet superseded the use of rotary instruments. Furthermore, the concept of micro-cavity preparation developed in recent years and the introduction of acid-etch techniques, resin bonding and the use of glass-ionomer cements have also revolutionised the principles of cavity preparation in conservative dentistry. This article reviews the development of these various caries removal techniques and instrumentation and the evolutionary philosophies of cavity preparation promulgated over the last century or so. PMID- 10388385 TI - Dentin demineralization inhibition at restoration margins of Vitremer, Dyract and Compoglass. AB - PURPOSE: To examine the in vitro caries inhibition of a resin-modified glass ionomer cement (Vitremer-3M) and two compomers (Dyract-Dentsply; Compoglass Ivoclar). MATERIALS AND METHODS: Standardized Class V preparations were placed in 40 molars, the gingival margin placed below the cementoenamel junction. Randomly, 10 Vitremer, 10 Dyract and 10 Compoglass restorations were placed according to manufacturer's instructions, in 30 teeth. Ten teeth received P-50 composite resin (3M) restorations and acted as the control. All teeth had an acid-resistant varnish placed to within 1 mm of restoration margins and they were then placed into artificial saliva for 4 weeks, the saliva being replenished every 48 hours. All teeth were subjected to an artificial caries challenge (pH 4.4) for 5 days. Sections of 100 microns were obtained, photographed under polarized light microscopy, and then digitized to quantitate demineralized areas adjacent to the restoration. RESULTS: The mean (+/- S.D) area (microns 2) demineralization 100 microns from the dentin/gingival margin was: Vitremer 4965 +/- 841, Compoglass 4981 +/- 2209, Dyract 5375 +/- 516, P-50 8088 +/- 2083. ANOVA and Duncan's (P < 0.05) indicated all three fluoride-releasing materials examined in this study had significantly less demineralization adjacent to restoration margins than the P-50 composite resin control. Seventy percent of glass ionomer cement restorations demonstrated adjacent dentin inhibition zones, while no dentin inhibition zones were demonstrated with the compomer restorations. PMID- 10388387 TI - Risk periods in the development of dental fluorosis. AB - In order to study the age-related susceptibility to dental fluorosis, 40 children who had been lifelong consumers of moderate- to high-fluoride water (0.55-8.48 mg F/l) were examined, as well as a group of older siblings (n = 40) who were born 6 months to 6 years before the fluoride-containing drinking water was introduced to the household. Background information was obtained through a structured questionnaire. Dental fluorosis was scored according to the TF index. Among the 80 children examined, the permanent incisors were erupted in 66, while 67 had permanent first molars present. As compared to their older siblings, the prevalence of dental fluorosis was significantly higher in the children who had consumed moderate-to high-fluoride water throughout their lives. In a multiple regression analysis, the variable "age when introduced to moderate- to high fluoride water" came out as the only significant risk factor associated with dental fluorosis. This variable was divided into three categories according to the first exposure to moderate- to high-fluoride drinking water (1) 0-12 months of age, (2) 13-24 months of age and (3) after 24 months of age. Category 3 was used as the reference group. Fluoride exposure starting during the 1st year of life showed the highest odds ratio as compared to exposure only after 2 years of age. The findings indicate that early mineralizing teeth (central incisors and first molars) are highly susceptible to dental fluorosis if exposed to fluoride from the first and--to a lesser extent--also from the 2nd year of life. PMID- 10388388 TI - Retrospective clinical investigation and survival analysis on ceramic inlays and partial ceramic crowns: results up to 7 years. AB - This study retrospectively evaluated the clinical performance of 287 all-ceramic restorations placed during routine patient care in the University setting in the past 7 years. All patients (n = 106) with ceramic inlays or partial ceramic crowns (PCC), placed during 1988-1994 (n = 327) by five experienced dentists were asked to take part in a clinical investigation, and 92 patients with 287 restorations (232 inlays, 55 PCC) agreed to do so. The following ceramics were used: 44 (15.3%) Dicor (Dentsply), 126 (43.9%) IPS-Empress (Ivoclar), 82 (28.7%) Mirage II, 33 (11.5%) Cerec-Vita-Mark 1 (Vita), and 2 (0.7%) Duceram LFC (Ducera) restorations. The restorations were placed using the following luting composites: 73 (25.4%) Dual Cure Luting Cement (Optec), 81 (28.3%) Variolink high viscosity (Ivoclar), 32 (11.1%) Microfill Pontic C (Kulzer), 51 (17.8%) Dual Zement (Ivoclar), 40 (13.9%) Dicor Light Activated Cement (Dentsply), and 10 (3.5%) Vita Cerec Duo Cement (Vita). Restorations were evaluated according to the modified USPHS criteria. Kaplan-Meier analysis was used to calculate the probability of survival. Of the 287 restorations 270 (94.2%) were still in function without any need of intervention. Fourteen restorations (4.8%) had failed before starting the clinical investigation, and in three a fracture was found during the investigation. These 17 failed restorations consisted of 14 PCC and 3 ceramic inlays. The results of the clinical investigation revealed 59.2% Alpha-ratings for marginal adaptation. Only one restored tooth showed recurrent caries. The probability of survival (95% confidence interval) for 7 years was 98% (97.99 98.01%) for ceramic inlays and 56% (46-66%) for PCC. Our findings show that ceramic inlays can be regarded as an acceptable alternative to cast gold restorations within the methodological limitations of the present study. For PCC further experience with more recent ceramics is warranted. PMID- 10388389 TI - Retention and marginal adaptation of a compomer placed in non-stress-bearing areas used with the total-etch technique: a 3-year retrospective study. AB - The aim of this clinical study was to evaluate class V and class III cavities restored with a polyacid-modified resin composite (compomer) restorative material in association with two different dentin-enamel bonding systems: Dyract-PSA (Primer Sealer Adhesive-DentSply, Germany) and Prime & Bond 2.0 (DentSply, Germany). The control group was a hybrid composite used with ProBond bonding system (DentSply, Germany). A total of 116 restorations (79 class V, 37 class III) were made and reevaluated after 1, 2 and 3 years in 55 patients in two private practices and in a university department. Class V nonretentive cavities were located at the CEJ level and class III at interproximal level close to CEJ. Each cavity was prepared using a water-cooled, high-speed handpiece with a fine diamond burr. A small bevel was prepared along enamel margin. Cavity dimensions were no more than 3.5 x 3.5 mm (using burr as reference point). Each restoration was finished immediately with fine diamond burrs and Sof-Lex disks (3 M, USA). The criteria that were evaluated by the USPHS method included: retention, color match, marginal integrity, marginal discoloration, and secondary caries. Results indicated that all compomer restorations were fully retained at 3 years, and that no secondary caries detected. Seven composite restorations were lost during the 3 year study. No statistical differences were observed between class III and class V or among other conditions (e.g., upper-lower arc, sex, age). This study demonstrates that compomers are suitable restorative materials for class III-V restorations. They may represent a clinical alternative to composites in class V and III restorations. PMID- 10388390 TI - Three-dimensional reconstruction of approximal subsurface caries lesions in deciduous molars. AB - The present study examines the three-dimensional (3D) morphology of early approximal subsurface enamel caries lesions and subjacent dentin reactions in deciduous molars. Twenty-three extracted primary molars were embedded in Technovit 9100 and serial sections were cut using a saw microtome. Forty approximal lesions were found and investigated using polarized light microscopy for the identification of the different zones of the caries lesion. These zones were then reconstructed three-dimensionally using computer-aided 3D reconstruction methods and the dimensions and volumes of the translucent zone, the body of the lesion, and the dentin lesion were calculated. The enamel demineralization index was defined as the ratio between the translucent zone and the body of the lesion, whereas the enamel-dentin demineralization index was defined as the volumetric ratio of the early dentin lesion to the body of the enamel lesion. The 3D reconstruction of the lesions showed extremely heterogeneous micromorphological features of zone profiles. In enamel lesions, the results demonstrated a decreasing enamel demineralization index with increasing size of the lesion, which indicates a high risk of further caries progression. The enamel-dentin demineralization index indicated, in 5 out of 17 dentin lesions, a high risk of further caries progression. Computer-assisted 3D reconstruction and volumetric assessment of initial caries lesions in deciduous molars represents a valuable methodology in pathogenesis studies, which may lead to a better clinical understanding of caries progression. PMID- 10388391 TI - Development of gingivitis around aged restorations of resin-modified glass ionomer cement, polyacid-modified resin composite (compomer) and resin composite. AB - Resin-modified glass ionomer cements (RMGIC) and polyacid-modified resin composites (PMC, compomers) are two recently introduced material groups supposed to replace traditional cements in operative dentistry. The new restoratives release initially fluoride in different relatively high concentrations, which decrease gradually during the first weeks in vivo. Earlier studies showed a stronger subclinical inflammatory reaction around different conventional tooth colored restorative materials than around intact enamel. The aim of this study was to compare intra-individually the initiation of gingival inflammation around, aged RMGIC, PMC and resin composite restorations. Subgingivally located Class III restorations were placed in 17 patients. Each patient received one of each of the experimental materials. All patients were placed on an oral hygiene regime 1-year after finishing of the restorations. Gingivitis was induced during a one-week period without oral hygiene. The gingival condition was assessed by sampling of gingival crevicular fluid (GCF), registration of the amount of bacterial plaque and by registration of bleeding after gentle probing of the entrance of the gingival sulcus (SBI) on the experimental filling- and control-enamel surfaces at days 0 and 7. No differences were seen in plaque and gingival index scores between the materials at both days. The GCF increased significantly for all surfaces during the experimental gingivitis period. At day 7 significantly lower GCF was sampled around the enamel surfaces. In conclusion, the differences between the materials did not result in measurable differences concerning clinical or subclinical signs of gingivitis. PMID- 10388392 TI - Interfacial adaptation of in vivo aged polyacid-modified resin composite (compomer) restorations in primary molars. A SEM evaluation. AB - Polyacid-modified resin composite (PMC) restorations are being increasingly used in class II cavities in primary teeth. The aim of this study was to evaluate the interfacial adaptation of 1-month- and 30-month-old in vivo restorations by quantitative scanning electron microscopy (SEM) evaluation. Twelve PMC restorations were performed under clinically controlled conditions in primary molars planned for extraction 1 month later for orthodontic reasons. Eleven other PMC restorations, aged 30 months (range 1.5-3 years) and part of a multicenter study, were collected after exfoliation. To observe the interfacial adaptation of each restoration at several levels a thin layer of the proximal surface was ground off 2-3 times. Replica impressions of each level were prepared for SEM. The interfaces of the replicas were evaluated at x200 and x1000. In the 1-month old restorations gap-free adaptation to enamel was found in 87% and to dentin in 84% of the total interfacial length investigated. For the 30-month-old restorations gap-free adaptation was registered in 59 and 63%, respectively. The interfacial quality was significantly better in the 1-month-old restorations than in those 30 months of age. Adaptation to enamel was significantly better in the cervical part of the 1-month-old restorations than in the axial walls, whereas there was no significant difference in dentin. No significant difference was found between the cervical and axial cavity walls of the 30-month-old restorations. Enamel fractures were registered in 31 and 24%, respectively of the interfacial length of the 1-month- and 30-month-old restorations. The corresponding findings in dentin were 0 and 0.9%. It can be concluded that restorations aged for a short time showed a high percentage of sealing, which decreased significantly for the 30-month-old ones. PMID- 10388393 TI - s-IgA and cytokine levels in whole saliva of Sjogren's syndrome patients before and after oral pilocarpine hydrochloride administration: a pilot study. AB - Previous investigations have found elevated levels of s-IgA in the parotid saliva and normal levels in submandibular saliva of patients with Sjogren's syndrome (SS). Fox et al. also found elevated levels of cytokines (i.e., IL-2 and IL-6) in serum, salivary epithelial cells and parotid saliva of patients with SS. The oral administration of pilocarpine hydrochloride stimulates whole and parotid salivary flow. The purpose of this study was to determine the levels of s-IgA and IL-2 and IL-6 in whole saliva before and after administration of pilocarpine hydrochloride in SS subjects. Ten definitively diagnosed SS subjects were enrolled in the study, as were ten controls (C). The mean age was 57.2 years and all subjects were female. Whole unstimulated saliva (WUS) was collected by standard techniques for 5 min, after which the volume and flow rate were determined (mean WUS: SS = 0.047 vs C = 0.480 ml/min). Samples were centrifuged and the immunoglobulin analysis performed on the supernatants by immunoreactivity in a double-sandwich technique as previously described by Rudney et al. Cytokine analysis was performed similarly utilizing commercially available kits from R&D Systems. The results as analyzed by pairwise t-tests revealed comparable levels of s-IgA in the saliva of the SS patients, as compared to controls at baseline (means +/- SEM: SS-IgA = 348.1 +/- 82.0 vs C-IgA = 284.0 +/- 65.1 micrograms/ml; NS). Whole salivary flow was significantly increased (328%) in the SS subject group 60 min after the administration of 5 mg pilocarpine hydrochloride (means +/- SEM: 0.0472 +/- 0.017 vs 0.1546 +/- 0.054 ml/min; P < 0.01). There was no significant change in the concentration of s-IgA in the SS subject group following the pilocarpine dose (means +/- SEM: SS-IgA = 439.9 +/- 121.2 microliters/ml; P = NS). There were elevated levels of IL-2 in the saliva of four out of the ten and IL-6 in two out of the ten SS patients, as compared to controls (means +/- SEM: SS-IL-2 = 127.8 +/- 11.4 vs C-IL-2 = 30.8 +/- 1.6 pg/ml and SS-IL-6 = 41.4 +/- 7.1 vs C-11.6 +/- 2.8 pg/ml). There was also a significant decrease in the concentration of IL-2 in the same four out of ten SS subjects following the pilocarpine dose (means +/- SEM: SS-IL-2 = 32.4 +/- 10.3; P < 0.01). These preliminary results indicate that s-IgA levels do not change with increased salivary flow following the administration of pilocarpine hydrochloride in patients with Sjogren's syndrome. While cytokines are elevated in the whole saliva of some SS patients, a decrease in IL-2 concentration may occur with increased salivary flow. PMID- 10388394 TI - Digital photography in the orthodontic practice. PMID- 10388395 TI - Ectopic eruption of mandibular incisors. PMID- 10388396 TI - Management service organization contracts: tips for successful negotiations. PMID- 10388397 TI - A rationale for removable retainers. PMID- 10388398 TI - The Damon low-friction bracket: a biologically compatible straight-wire system. PMID- 10388399 TI - Maintaining anchorage with a combination Nance-Goshgarian transpalatal arch. PMID- 10388401 TI - Where are the assistants? PMID- 10388400 TI - Marking patient casts. PMID- 10388402 TI - The Connecticut Intrusion Arch. PMID- 10388403 TI - Segmental lingual orthodontics in preprosthetic cases. PMID- 10388404 TI - The effectiveness of an elastomeric module dispenser in cross-infection control. PMID- 10388405 TI - Orthodontic fees. PMID- 10388406 TI - Clinical use of the Churro jumper. PMID- 10388407 TI - Treatment of mandibular alveolar prognathism by a lower anterior subapical osteotomy. PMID- 10388408 TI - An outlook on the numbers game: a qualitative assessment. PMID- 10388409 TI - Increased intracranial pressure after coronal suturectomy in craniosynostotic rabbits. AB - It has been suggested that the complications associated with intracranial hypertension in craniosynostotic infants may be managed with surgical release of the synostosed sutures. However, both postoperative increases and decreases in intracranial pressure (ICP) have been reported in heterogeneous samples of infants with syndromic and nonsyndromic craniosynostoses. The present study was designed to describe longitudinal changes in ICP in a homogeneous sample of rabbits with uncorrected and corrected familial coronal suture synostosis and compare them with age-matched normal control rabbits. Fifty-three rabbits were divided into three groups: normal rabbits (n = 28), rabbits with uncorrected bilateral coronal suture synostosis (n = 9), and rabbits with bilateral coronal suture synostosis with coronal suturectomy at 25 days of age (n = 16). ICP was measured at 25 and 42 days of age using a Codman epidural microtransducer. Results revealed that rabbits with uncorrected craniosynostosis had significantly (P < 0.05) higher ICP at 25 days of age than normal control rabbits by approximately 86%. However, by 42 days of age, ICP in normal rabbits increased by 75%, whereas ICP in rabbits with uncorrected craniosynostosis decreased by 69% over the same time. Synostotic rabbits with coronal suturectomy showed a 50% decrease in ICP immediately after surgical release and then followed the normal, age-related ICP pattern, which significantly increased by 75% at 42 days of age. Results suggest that, in the rabbit model, the postsuturectomy rise in ICP may simply be normal, age-related changes, although a longer follow-up will be needed to determine the recurrence of pathological ICP. Possible multifactorial explanations for intracranial decompression and compensation in the craniosynostotic rabbit model are also discussed. PMID- 10388410 TI - Craniofacial correction of giant frontoethmoidal encephalomeningocele. AB - The surgical treatment of a very large anterior encephalocele in an infant is presented. Because of the large size of the encephalocele, a combined transfacial transcranial approach was used for correction of the associated intracranial, cranioorbitonasal bone, and facial skin deformities. PMID- 10388411 TI - Maxillary distraction in cleft lip palate patients: a review of six cases. AB - Cleft lip and palate patients can present with a maxillary retrusion with tendency to Class III malocclusion after cleft repair. Maxillary distraction osteogenesis is a technique that provides simultaneous skeletal advancement and expansion of the soft tissues. Six nonsyndromic cleft lip and palate patients, ages 12 to 16 years (mean, 13.8 years), underwent maxillary distraction; four had a unilateral and two a bilateral cleft lip and palate. After an incomplete LeFort I osteotomy; a latency period of 3 days was respected. On Postoperative Day 4, distraction was initiated through anterior traction on a Delaire facial mask using distraction forces of 900 gm. Photographs and lateral cephalometric radiographs were obtained preoperatively and 4 months after distraction. A cephalometric analysis was performed to compare the sagittal dentocraniofacial morphology before and after distraction. The aesthetic improvement obtained by maxillary distraction osteogenesis during the permanent dentition to correct maxillary retrusion in our cleft lip and palate patients was impressive. Skeletal advancement varying from 1 to 3.5 mm (mean, 1.7 mm) was found. However, significant dentoalveolar compensations occurred in three patients. This was due to the dental anchorage of the distraction device and can be avoided only by the use of skeletal fixation. PMID- 10388412 TI - Distraction osteogenesis of free interpositional membranous bone: experimental design. AB - The purpose of this study is to explore the possibility of bone formation in distraction osteogenesis of the free interpositional membranous bone. Three canine dogs were used as subjects. Two mandibular osteotomies were made in the mandibular body. The oral soft tissue and periosteum of the segment between the two osteotomies were freed from periosteum and the surrounding soft tissue as a free interpositional bone graft 2 cm wide was created. The free interpositional bone was fixed to the proximal mandibular body with a miniplate. The external fixation device was applied to the proximal mandibular body and the distal mandibular body. Mandibular distractions were performed postoperatively at a rate of 1 mm/day for a total of 10-mm distraction for 10 days. The latency period was 1 week in Dog 1, 2 weeks in Dog 2, and 3 weeks in Dog 3. Three dogs were killed 6 weeks after distraction, and interpositional bone specimens were obtained. In Dogs 1 and 2, the free interpositional bone showed severe resorption and had no new bone formation at the distracted area. However, in Dog 3, new bone developed along the distracted gap. Our study demonstrated the possibility of distraction osteogenesis in the free interpositional membranous bone and suggested that free interpositional membranous bone be allowed, under rigid fixation device, to have enough revascularization from surrounding tissue to have osteogenesis for at least 3 weeks or even more. PMID- 10388413 TI - Surgical anatomy of the orbit of Korean adults. AB - When operating in and around the orbit, the key to a successful result is precise anatomical localization. However, there is no precise study about the localization of vital orbital structures from reliable periorbital bony anatomy of the Korean adult. This study was constructed to give pertinent anatomical measurements to which the plastic and maxillofacial surgeon may refer. The 82 orbits obtained from 41 skulls of adult Koreans were measured with Vernier calipers and Marshac calipers. Superiorly, the supraorbital fissure was 40.0 +/- 2.5 mm from the supraorbital notch. Medially, the posterior ethmoidal foramen was 31.7 +/- 3.0 mm from the anterior lacrimal crest. Inferiorly, the infraorbital fissure where the infraorbital groove started was 26.4 +/- 2.6 mm from the infraorbital foramen. Laterally, the supraorbital fissure was 34.3 +/- 2.7 mm from the frontozygomatic suture. These distances are suggested as appropriate safe distances from each periorbital bony landmark. Dissection beyond that distance should be done with great caution. PMID- 10388414 TI - Management of frontoethmoidal (sincipital) encephalocele. AB - Frontoethmoidal encephaloceles are congenital malformations that cause complex deformities in the frontal, orbital, and nasal regions. As the term implies, with frontoethmoidal encephaloceles, intracranial material has herniated through the dural and skull defect. In this report, 21 patients with frontoethmoidal encephalocele operated by a craniofacial team are presented, and accompanying anomalies, results, and complications are discussed. PMID- 10388415 TI - Retrospective study of nonsyndromic craniosynostosis treated over a 10-year period. AB - Since the first operation for premature suture closure in North America in 1888, there have been some fundamental changes in the treatment of these sutures, the latest being the U.S. Food and Drug Administration's 1996 approval of a bioabsorbable fixation device. This retrospective study documents our experience with procedures performed primarily by Bennie J. van R. Zeeman for isolated craniosynostosis over a 10-year period. It was an attempt to evaluate factors affecting outcome and to determine the safety of the techniques used to correct these congenital defects. Diagnoses included plagiocephaly (116) and sagittal (44), metopic (17), and bilateral coronal (12) synostosis. All patients underwent fronto-orbital advancement or calvarial vault remodeling, or both. The average patient age at time of sagittal synostosis surgery was 13.4 months; unilateral coronal synostosis, 12.2 months; deformational plagiocephaly, 9.8 months; metopic synostosis, 8.6 months; and bilateral coronal synostosis, 10.4 months. Perioperative complications were minimal, with one mortality. Postoperative complications included three cases involving infection. The problem of reoperation for the removal of wires and plates remains the greatest postoperative complication. Because of poor patient compliance, no accurate postoperative follow-up has been recorded. On the basis of our experience, we wish to point out some problems inherent in this surgery and also the complications that can occur despite careful coordinated planning and team effort. PMID- 10388416 TI - Spontaneous closure of bony defect in a frontoethmoidal encephalomeningocele patient. AB - The frontoethmoidal encephalomeningocele (FEEM) is a congenital herniation of meninges and brain tissue through the skull bony defect at the foramen cecum. The size of the defect may vary from a few millimeters to many. Those patients with a small defect may not always require a risky operation during childhood. We report on an infant whose bony defect has closed spontaneously with definite clinical evidence. It is proved that the skull defect and brain herniation are able to heal naturally, and this affirms an existence of the abortive subtype of FEE. Conservative treatment may be considered in those with a small bony defect, and surgery can be considered later when it is required. PMID- 10388417 TI - Orthognathic surgery in craniomaxillofacial fibrous dysplasia. AB - In craniomaxillofacial fibrous dysplasia, jaw involvement often causes facial asymmetry, an occlusal cant, and loss of teeth. Although conservative management of fibrous dysplasia affecting the jaws is widely practiced, orthognathic surgery is indicated in such cases to restore occlusion and correct dentofacial deformity brought on by the disease process. Since 1981, the Craniofacial Center at Chang Gung Memorial Hospital in Taiwan has treated a total of 84 patients with craniomaxillofacial fibrous dysplasia. Of these, 55 (65%) had fibrous dysplasia affecting the jaws (Zone 4). Between 1988 and 1997, orthognathic surgery was performed on 1 male and 4 female patients with fibrous dysplasia involving the teeth-bearing jaws. One patient had localized fibrous dysplasia that involved the mandible. The other 4 patients had polyostotic craniofacial involvement of Zones 1, 2, or 3 and 4A. The patient with isolated mandibular involvement and 2 patients with maxillary fibrous dysplasia had single-jaw surgery. The other 2 patients with maxillary involvement required simultaneous two-jaw surgery to correct the dentofacial deformities resulting from the disease process. Follow-up ranged from 12 months to 9 years. All the patients had stable occlusion, good facial aesthetics, and no further recurrence after surgery. The long-term stability of the achieved occlusion and facial appearance confirms that adequate healing in fibrodysplastic bone is to be expected using the standard fixation. PMID- 10388418 TI - Hallermann-Streiff syndrome: experience with 15 patients and review of the literature. AB - Hallermann-Streiff syndrome is rare, with approximately 150 case reports in the world literature. The syndrome consists of proportionate nanism; hypotrichosis; atrophy and extreme thinness of the skin, particularly over the facial area; an unusual "bird-like" face with mandibular hypoplasia; a prominent thin, pointed nose; congenital cataracts; and severe dental abnormalities. This appears to be a sporadic mutation, and the inheritance pattern is unknown. Clinical management must focus on the more life-threatening and developmental issues early on, and aesthetic deformities can be addressed after the adolescent growth period is complete. Surgical correction of cataracts should be undertaken early in life to preserve vision. Airway issues need to be addressed early. Other reconstructive procedures, including rhinoplasty, facial augmentation, and mandibular surgery, have been successful and can be performed later in life. We report on our clinical findings in 15 patients with this condition, our attempts at reconstruction, and complications we have encountered in treating this patient population. Five of our patients had produced normal chromosome studies, and none have had similarly affected siblings. Four have had normal, unaffected children. Most of our patients have undergone multiple reconstructive procedures and have done relatively well. Eleven of our patients, however, have encountered significant intermittent respiratory difficulty manifested as early feeding difficulty, recurrent upper respiratory tract infection, sleep apnea, and respiratory arrest. Three patients required tracheostomy because of respiratory difficulty, and one child died of postoperative respiratory compromise. The management of these complicated and difficult patients is discussed. PMID- 10388419 TI - The brave new digital world. PMID- 10388420 TI - Analysis of workforce, distribution of care, and practice preference in pediatric plastic surgery. AB - To determine the future needs in manpower for pediatric care as it relates to pediatric specialists, a study was conducted by the American Academy of Pediatrics to see the needs of manpower that will provide access of pediatric care to all. A pediatric plastic surgery survey was set in the form of a list of questions that was mailed to the respective societies with pediatric plastic surgeons as members. The survey was reviewed, and the results were studied. The outcome is presented in the form of findings related to the overall practice of plastic surgery. Based on the percentage of pediatric care that is provided, there were two types of pediatric plastic surgeons. Those with the high percentage of pediatric care tend to stay near health science centers; however, both groups tend to spend time (each to a different extent) tending to other plastic surgery problems. Today we have adequate access to care in the health system for pediatric plastic surgery problems despite the shift in the health care environment. Managed care continues to use the pediatrician as a "gatekeeper" in determining the overall access for patients with problems related to pediatric plastic surgery. PMID- 10388421 TI - Correction of the contracted eye socket and orbitozygomatic hypoplasia using postauricular skin flap and temporal fascial flap. AB - The authors corrected a contracted eye socket and orbitozygomatic hypoplasia simultaneously, secondary to previous surgery and radiotherapy. A one-stage surgical reconstruction was undertaken using both a postauricular skin flap and a temporal fascial flap, which were rotated for eye socket reconstruction and bone graft coverage. Hydroxyapatite was inserted as an onlay bone graft substitute to enlarge the orbitozygomatic region. The authors' experience with 12 patients resulted in contours that were satisfying and encouraging. f1 PMID- 10388422 TI - Comparison of anthropometric and cephalometric measurements of the adult face. AB - To tabulate and assess quantitative differences between anthropometric and corresponding radiographic cephalometric measurements obtained from the same persons, 19 projective linear measurements were taken from the surface of the heads and faces of 41 patients with cleft lip, cleft palate, or both, all of whom were white North Americans aged 14 to 29 years. They underwent radiographic examination shortly afterward, and corresponding cephalometric measurements were obtained. Differences between the methods were assessed by the numeric differences between the mean values of concurrent measurements. Statistical difference was assessed by paired t test, Pearson product-moment correlations, and intraclass index for degree of agreement between findings. By numeric difference, 6 of 19 anthropometric measurements were similar to (within 2% of) those taken from radiographs. Paired t tests disclosed significant differences between 16 of 19 measurements. Half of the six surface measurements similar to their skeletal counterparts showed no statistical difference; the other half showed only moderately significant differences. Differences between the 13 "dissimilar" measurement pairs (differences > 2%) were highly significant. Good correlations were found in five of the six similar measurements, which may have an important prognostic value in understanding changes in the craniofacial measurements of the face. Knowledge of the correlations between all major measurements of the head and face on the surface and skeleton is essential for anticipating changes in the morphologic characteristics of the growing face. PMID- 10388423 TI - Coronal suture response to distraction osteogenesis in rabbits with delayed-onset craniosynostosis. AB - Recent studies have identified a subpopulation of persons with craniosynostosis who exhibit progressive or delayed-onset synostosis and mild cranial vault deformities. These persons may be good candidates for nonextirpation distraction osteogenesis. The present studies were designed to determine force-displacement parameters and assess the effects of distraction osteogenesis on coronal suture growth and morphologic characteristics in a rabbit model with congenital, delayed onset craniosynostosis. Data were collected from a total of 178 rabbits: 71 normal controls; 16 normal controls with distraction; 72 with delayed-onset coronal suture synostosis; and 19 with delayed-onset coronal suture synostosis and distraction. At 10 days of age, all rabbits had amalgam markers placed on both sides of the coronal suture. In the force-displacement study, force displacement distractors were placed across the coronal suture and distracted acutely for 1.0 mm at 42 days of age. Force-displacement curves for the coronal suture were best described by a third-order polynomial regression equation for both normal and synostosed groups. Significant differences (P < 0.05) were found in the mean force necessary to distract a normal suture 1 mm in distance (13.72 kg) compared with a suture with delayed-onset synostosis (48.39 kg). A significant (P < 0.05) relationship was also found between the extent of synostosis and the distractive force in rabbits with delayed-onset synostosis. In the distraction study, internal distractors were fixed across the coronal suture at 25 days of age and percutaneously and intermittently activated at an average of 0.11 mm/day for 42 days (4.54 mm total). Serial radiographs were taken at 10, 25, 42, and 84 days of age. Results revealed that rabbits with delayed-onset synostosis and distraction had significantly (P < 0.01) more coronal suture growth rates compared with rabbits with delayed-onset synostosis and no distraction. Coronal sutures were harvested at 84 days of age for qualitative histologic examination. Normal, distracted coronal sutures showed widened sutural ligaments and thin, active osteogenic fronts. In contrast, distracted coronal sutures from rabbits with delayed-onset synostosis showed narrowed sutural ligaments, thickened and blunt osteogenic fronts, and increased collagen and bony matrix deposition compared with controls. Results suggest that distraction osteogenesis without corticotomy may be a treatment alternative in persons with progressive, delayed-onset synostosis. However, these preliminary data also suggest that distractive forces may accelerate or stimulate osteogenesis differentially in persons with craniosynostosis, possibly through an underlying genetic disorder of bone and cytokine regulation. These differential osteogenic responses to distraction, if validated clinically, will need to be taken into account when planning distraction rate and rhythm protocols for patients with craniosynostosis. PMID- 10388424 TI - Positron emission tomography studies confirm the need for early surgical intervention in patients with single-suture craniosynostosis. AB - Craniosynostosis, the premature fusion of one or more cranial sutures, may occur in isolation or in association with a syndromic constellation. Multiple-suture synostosis has consistently been associated with brain compression and increased intracranial pressure, and frequently decreased cognitive development. Single suture craniosynostosis, however, has been thought by some to be an aesthetic problem with infrequent consequences on brain function and development. Some studies have disputed this concept and have argued a correlation between single suture craniosynostosis and abnormalities in development. The purpose of this study was to determine, using an objective radiographic tool, positron emission tomography scans, if patients with single-suture craniosynostosis had any abnormalities in cerebral glucose metabolism that would indicate changes in local brain function. A total of 10 children with single-suture craniosynostosis, eight males and two females, ranging in age from 0.1 to 3.2 years, were enrolled in this prospective study approved by the Internal Review Board. Six of the children had sagittal synostosis, three had metopic synostosis, and one had coronal craniosynostosis. Each of the patients had preoperative positron emission tomography scans performed 1 to 5 weeks before cranial reconstructive surgery and postoperative scans at 6 to 12 weeks after surgery. Surgical treatment consisted of cranial vault remodeling in eight of the children and strip craniectomy with cranial expansion in two of the children. After surgery, the two scans were compared qualitatively and quantitatively by a single radiologist. The results demonstrated variable regional increases and decreases in local post-operative cerebral glucose metabolism. However, in the posterior occipital region, the area of visual development and visual spatial coordination, there was a consistent postoperative increase in all 10 patients. Maximum glucose metabolic rate was increased up to 30.2% with a mean of 9.9%, and average glucose metabolic rates demonstrated a maximum increase of up to 18.8%. The results of this study indicate cerebral glucose metabolism consistently increases in the posterior occipital cortex after surgical release of single-suture craniosynostosis. Future developmental studies are being performed to examine the functional consequences of these metabolic changes. PMID- 10388425 TI - Commentary on avoidance of implicit hazards: the realignment of maxillary and mandibular arches in comminuted and facial fractures. PMID- 10388427 TI - A management strategy for palatal fractures: a 12-year review. AB - Fractures of the palate are frequently associated with the more common and well described Le Fort fractures. Palatal fractures may present diagnostic and exposure challenges and, if not satisfactorily treated, will result in occlusal problems after surgery. From 1986 through 1998, 116 complex maxillary fractures were treated at the authors' center. Among these, 13 patients were diagnosed with fracture of the palate. Patients with gunshot wounds to the face were excluded from the present study. Open reduction and internal fixation of the palatal fractures were achieved through elevation of the entire palatal mucoperiosteal flap to avoid late hardware exposure. This paper presents a unique approach to visualizing the whole bony palatal surface for accurate reduction and internal fixation of fractures. PMID- 10388426 TI - Prophylactic use of ondansetron for emesis after craniofacial operations in children. AB - Children who undergo craniofacial operations are especially at risk of postoperative nausea and vomiting. These operations are more complex than the craniotomies for resective procedures. Postoperative vomiting is a common occurrence that can delay recovery and result in cerebrospinal fluid leak and fistula formation in these patients. Ondansetron, a selective serotonergic antagonist, is effective in reducing postoperative nausea and vomiting in several high-risk populations. In a randomized, double-blind, placebo-controlled study, the authors compared the prophylactic use of intravenous ondansetron 0.15 mg/kg with induction of anesthesia versus a placebo of normal saline 0.3 ml/kg with induction. A second dose was given 8 hours after the first dose. After surgery, episodes of vomiting were recorded separately in 0 to 2 hours, 2 to 6 hours, 6 to 12 hours, 12 to 24 hours, and 24 to 48 hours. Postoperative vomiting is significantly reduced in the ondansetron group compared with the placebo group (P = 0.000258). Ondansetron is effective in the prevention of postoperative vomiting in the pediatric population undergoing craniofacial operations. PMID- 10388428 TI - Primary alveolar cleft bone grafting in unilateral cleft lip and palate: arch dimensions at age 8. AB - The purpose of this investigation is to determine whether primary alveolar cleft bone grafting in infants with unilateral cleft lip and palate (N = 17) leads to less favorable dental arch dimensions at age 8 when compared with other 8-year old patients with unilateral cleft lip and palate who received no alveolar bone grafting procedures (N = 49). Dental casts were obtained for the primary grafted group, and arch lengths and widths were digitally recorded with a reflex microscope. These arch dimensions were then compared with the reported data for a nongrafted group and a noncleft group of 8-year-old children. The major findings were: 1) that the dental arches of both cleft groups generally demonstrated a significant diminution in length and width (P < 0.05) compared with the noncleft groups, and 2) that the patients who underwent primary alveolar cleft bone grafting showed no statistically significant difference for any arch dimension (P < 0.05) when compared with the nongrafted group lacking this additional surgical procedure. PMID- 10388429 TI - Primary alveolar cleft bone grafting in unilateral cleft lip and palate: craniofacial form at age 8. AB - Counterpart analysis can be advantageous for the clinician interested in the underlying determinants of the craniofacial form for any given person. This analysis was performed for a group of patients who underwent primary alveolar cleft bone grafting (N = 18) and a group of patients who did not undergo grafting (N = 19) who were 8 years of age (+/- 6 months). The primary grafting group more frequently noted maxillary retrusion, but of a nonsignificant magnitude. Also, the primary grafting group had greater mean magnitudes of mandibular opening as a compensatory adjustment in some patients, but this could not be generalized to all patients who had underdone primary grafting. The mean magnitude of craniofacial vertical shortening was also greater for some patients who had undergone primary grafting, but it, too, did not exhibit a generalized pattern for all patients who had undergone primary alveolar cleft bone grafting procedures. This study emphasizes the great diversity of craniofacial skeletal adjustments made within each group of patients with unilateral cleft lip and palate and cautions the clinician against generalizations concerning a particular treatment protocol. PMID- 10388430 TI - Stage grouping of oropharyngeal cancer: evaluation of three systems by means of survival analysis. AB - Stage grouping is a method of summarizing multiple categories generated from the tumor, node, metastasis (TNM) classification system. Recently three different systems, the T and N integer score (TANIS), Hart, and 1997 Union Internationale Contre le Cancer systems, have been proposed. To verify their correlation to survival, a series of 64 patients affected by primary squamous cell carcinoma of the oropharynx was considered in this retrospective study. The data set was classified according to Union Internationale Contre le Cancer T stage and then grouped as recommended by the three systems. Results showed a crude survival rate of 28.13%. Univariate analysis by means of the logrank test yielded significant P values for the T and N integer score and Hart systems (0.0452 and 0.0179, respectively) and a borderline P value (0.0728) for the stratification based on the 1997 Union Internationale Contre le Cancer system. Multivariate analysis (Cox regression adjusted for age and sex) showed a significant correlation between the three staging systems and the mortality rate. Odds ratios were 1.36 (95% confidence interval, 1.12-1.66), 1.58 (95% confidence interval, 1.18-2.12), and 1.63 (95% confidence interval, 1.08-2.48) for the T and N integer score, Hart, and 1997 Union Internationale Contre le Cancer systems, respectively. The T and N integer score system showed the best statistical correlation, but a conclusive result could not be achieved because of the low number of patients in this study. PMID- 10388431 TI - The role of transforming growth factor-beta, insulin-like growth factor I, and basic fibroblast growth factor in distraction osteogenesis of the mandible. AB - Distraction osteogenesis is a viable method for regenerating large amounts of bone. In contrast to fracture healing, the mode of bone formation in distraction osteogenesis is primarily intramembranous ossification. The basic biology of the process is still not well understood. The growth factor cascade is likely to play an important role in distraction. This study examines the growth factor cascade in a lengthened ovine mandible model. Twenty-four animals were divided into four groups with varying rates of distraction (1, 2, 3, and 4 mm/day). A unilateral distractor at the angle of the mandible was used. The mandibles were lengthened to 24 mm and fixed for a period of 5 weeks, after which the animals were killed. The sections were probed for transforming growth factor-beta, basic fibroblast growth factor, and insulin-like growth factor I. The growth factors studied were present in all four groups. Transforming growth factor-beta, basic fibroblast growth factor, and insulin-like growth factor I were present in both the bony matrix of the sections and the cytoplasm of the cells, osteoblasts, and a small number of mesenchymal cells. The sections obtained from groups distracted at faster rates showed stronger presence of the growth factors examined by more intense staining. In fracture healing, the localization of transforming growth factor-beta in stage I of healing corresponded with the precise region of intramembranous ossification in stage II. Diffuse presence of transforming growth factor-beta throughout the lengthened region corresponded with the process of intramembranous ossification observed in distraction. In fracture healing, insulin-like growth factor I and basic fibroblast growth factor have been shown to promote proliferation and differentiation of osteoblasts from precursor cells. The intense presence of insulin-like growth factor I and basic fibroblast growth factor in the distracted region may account for osteoblast proliferation and formation from precursor mesenchymal cells. Mechanical strain has been shown to increase the expression of transforming growth factor-beta and insulin-like growth factor I. Distraction may serve as a source of mechanical strain, which may explain, in part, the expression of these growth factors, particularly in the faster groups. PMID- 10388432 TI - Condylar fractures during growth: follow-up of 16 patients. AB - Fractures of the mandibular condyle represent 20% to 35% of all mandibular fractures. There are several clinical variants of this type of fracture that give rise to different problems in relation to their classification and treatment. A sample of 16 patients (of a total of 280 patients examined and treated from 1985 through 1995) with mono- and bilateral, displaced and decomposed, condylar fractures that occurred during growth were examined by the authors, who assessed, by a 2-year follow-up, the relevant clinical, functional, and instrumental parameters. On the basis of the data gathered by this study, a plan was drawn up for treating these patients that takes into account the different situations, such as either a nonsurgical or surgical treatment (by the use of condylectomy or external rigid fixation), and points out the advantages and disadvantages of each method. PMID- 10388433 TI - Moebius syndrome: the new finding of hypertrophy of the coronoid process. AB - The first detailed description of congenital facial paralysis was reported by Moebius in 1888. It is characterized by either unilateral or bilateral paralysis of the facial muscles and an associated abducens palsy. The present report is of two patients with Moebius syndrome, who were also diagnosed with trismus at birth. Each patient also demonstrated bilateral hypertrophy of the coronoid process of the mandible. In effect, the zygoma obstructed the excursion of the mandible because of a "coronoid block." A three-dimensional computed tomography scan demonstrated normal temporomandibular joints but bilateral hypertrophy of the coronoid processes and micrognathia. Both patients demonstrated less than 10 mm of oral excursion. Bilateral coronoidectomies were performed through an intraoral approach. The oral excursions after surgery increased to at least 20 mm. In each of these patients, the coronoid process was enlarged relative to the zygoma, which was of normal size and configuration. The trismus was associated with blocking of the coronoid by the anterior zygoma, preventing open or full excursion of the hypoplastic mandibles. Moebius syndrome can have a variable presentation at birth. In two patients, the authors describe a new finding of hypertrophy of the coronoid process and trismus secondary to obstruction of the coronoid by the hypertrophic zygomas during oral excursions. Each patient is described, and a review of the literature is discussed. PMID- 10388434 TI - The CDA family. PMID- 10388435 TI - Challenges and opportunities. PMID- 10388436 TI - Impaired dentists have a new place to find help. PMID- 10388437 TI - Dental management service organizations (DMSOs). PMID- 10388438 TI - New treatments hold perio promise. PMID- 10388439 TI - An oasis of development. PMID- 10388440 TI - 21st century dentistry. PMID- 10388441 TI - Microscope-assisted precision (MAP) dentistry. A challenge for new knowledge. AB - The purpose of this paper is to acquaint the dentist with the learning needs associated with the use of microscope-assisted precision dentistry (MAP) and identify certain concepts that will assist in the education process. Although the learning curve is considered to be lengthy and often difficult, the authors believe that the clinical benefits of MAP dentistry are well worth the necessary effort to achieve a level of competency with this methodology. PMID- 10388442 TI - The integration of filmless radiology in a restorative general practice. AB - A new faster, safer era has evolved for radiology in the dental office. This paper is a review of the latest technology that enables dentists to integrate filmless radiology into their practices. PMID- 10388443 TI - Digital tools for clinical dentistry. An Internet tutorial. AB - Computer technology has expanded the scope of digital tools that play a significant role in the practice of dentistry. In the search for current information and references about these devices, the Internet has become an essential resource. This paper will discuss some of the available digital adjuncts and their useful application in the clinical practice of dentistry, and will point the reader to web sites that are pertinent and informative. PMID- 10388445 TI - Current views on periodontal therapy. PMID- 10388444 TI - Children reap benefits from one man's desire to help. PMID- 10388446 TI - Periodontal regeneration: myth or reality? AB - One of the goals of periodontal therapy is regeneration. During the past 20 years, several materials and techniques have been developed and tested for enhancing periodontal regeneration. This paper evaluates flap debridement, allogenic and alloplastic grafting, and the use of nonresorbable and resorbable barrier membranes as regenerative techniques. One of the most predictable regenerative therapies is treatment of the three-walled intrabony defect. This defect can be repaired with 2 to 2.5 mm of bone fill and results in significant gains in clinical probing attachment and decreases in probing depths. There is a slightly greater improvement in periodontal measures with barrier membranes. Commercial preparations of allogenic bone and alloplastic fillers have a long, safe history of use and are primarily osteoconductive. They decrease probing depths and provide short-term gains in clinical attachment levels. Barrier membranes provide short-term evidence of improving Class II furcation invasions, however there is insufficient evidence that these improvements are sustained long term. Class III furcations are not predictably treated by regenerative therapies. To date, there is an absence of clinical evidence that regenerative therapy increases the long-term life span of teeth. PMID- 10388447 TI - A review of osseous resective surgery. AB - The treatment of periodontal diseases associated with attachment loss has involved a variety of approaches. While the goal of periodontal surgical treatments is to access the root surfaces for proper debridement, the decision to remove or reshape the supporting bone has been controversial. This paper will address the controversy as well as discuss surgical pocket therapy directed toward pocket reduction through recontouring the underlying bone. PMID- 10388448 TI - Periodontal medicine: assessment of risk factors for disease. AB - The approach to the diagnosis and treatment of periodontal disease is changing. The disease has not changed, but dentistry's understanding of the pathogenesis and appreciation for the influence of host factors has improved. As a result, the approach to the management of the disease is evolving. This paper reviews some of the host risk factors that have been linked to an increased severity of periodontal disease and briefly highlights some of the evidence that has led to the current belief that periodontal disease may be a risk factor for adverse systemic health conditions. PMID- 10388449 TI - Crown lengthening surgery: a restorative-driven periodontal procedure. AB - Improper management of the periodontal tissues during restorative procedures is a common, but often overlooked, cause of failure. When a restoration is placed, the preservation of an intact, healthy periodontium is necessary to maintain the tooth or teeth being restored. Predictable long-term restorative success requires a combination of restorative principles with the correct management of the periodontal tissues. PMID- 10388451 TI - Dentures made of Sevres Porcelain. PMID- 10388450 TI - Dentistry on stamps. PMID- 10388452 TI - Hal Harrison Ramsey: pioneer Texas dentist of the Big Country. AB - Smaller cities and rural communities often have limited information regarding their early health care practioneers. This is even more evident for dentists, especially those on the frontier. The obstacles those dentists overcame while practicing their profession are interesting by today's standards and their historical contributions to the profession and their local communities are worth preserving. Hal Harrison Ramsey was a circuit riding dentist during the settlement days of the West Central region of Texas often referred to as the Big Country by its present day residents. His story begins in the 1880s and covers the evolution of dentistry practiced by preceptor trained individuals to a profession of graduates from accredited dental institutions of the early 20th century. A legacy of dentists and physicians have continued through each generation of the Ramsey family and will do so into the 21st century. Dr. Ramsey's dentistry exemplifies the profession as practiced on the frontier of the late 19th century, and the heritage of contributions to the Big Country by his family are noteworthy. PMID- 10388453 TI - Driving out the 'toothworm' in today's China. AB - For thousands of years people have tried to explain toothache as the result of a worm residing in a carious tooth. Fumigation of the carious tooth, with the smoke from burning henbane or leek seeds, was used to drive the "toothworm" out of the tooth. A quack in modern China adopted the same ruse and supposedly drove the worms out of student's tooth. This duplicitous procedure was captured on film by a dentist who witnessed it. PMID- 10388454 TI - Dentistry in the literature of the XVIth and XVIIth centuries. AB - It is possible to get more than a glimpse of the dental condition of the English population in the XVIth and XVIIth centuries just by reading the works of the writers of the Elizabethan, Jacobean, and Caroline ages. Several dental references are included. PMID- 10388455 TI - Early dental instruments which are still in use today. PMID- 10388456 TI - Gleanings about dentistry from the world of literature (seventeenth in a series). PMID- 10388457 TI - [Chronic HCV hepatopathy and cryoglobulinemia. The associated clinical spectrum]. AB - BACKGROUND AND AIM: The hepatitis C infection (HCV) has numerous extrahepatic manifestations owing to the systemic nature of the infection itself. HCV infects the cells that carry a CD 81 receptor and show a marked tropism for hepatocytes, bone marrow staminal cells and circulating lymphomonocytes. One consequence of this tropism is the activation of B lymphocyte clones with the consequent production of autoantibodies and cryoglobulins. The secondary event is the formation of circulating immune complexes which, having precipitated at an intravascular level, may cause part of the extrahepatic manifestations associated with these infections. METHODS: This retrospective study evaluated the manifestations correlated and/or associated with HCV hepatitis and mixed cryoglobulinaemia. RESULTS: This analysis showed that 75% of consecutively studied patients reveal clinically important extrahepatic manifestations. CONCLUSIONS: This underlines the "broad spectrum" action played by the hepatitis C virus in the host organism. PMID- 10388458 TI - [Results of a survey on knowledge and habits among hospital doctors concerning primary prevention of venous thromboembolism]. AB - BACKGROUND: The purpose of this study was to analyse the attitudes towards and knowledge of primary prevention of venous thromboembolism (VTE) among hospital doctors in Calabria. METHODS: The survey was based on theoretical knowledge and practical management of hortopedics, surgeons, gynecologists and internists working in 14 hospitals. RESULTS: Out of a total of 340 physicians contacted, 154 (45%) agreed to take part in the survey. 82% of those who responded used VTE prophylaxis on a routine basis. Unfractioned heparin (71%) was the most frequently used methods; early deambulation (55%), low molecular weight heparin (49%) and elastic stocking (49%) were less frequently employed. Surprisingly, one third used aspirin. 75% of those contacted had modified their approach to prevention during the last few years, in particular owing to improvements in pharmacological therapy and increased awareness of the problem. In the survey of clinical practice, 80% of those who took part correctly identified the VTE risk, 86% suggested the best treatment, but only 27% assessed the exact frequency rate of deep venous thrombosis (DVT) and pulmonary embolism (PE) in the absence of prophylaxis. CONCLUSIONS: Most of the doctors contacted showed scant interest in the primary prevention of VTE. The 45% who agreed to be interviewed revealed a good practical approach but were not sufficiently aware of the real incidence of DVT and PE in a clinical risk context without prophylaxis. Although they must be interpreted with caution, these findings allow the real behaviour of hospital physicians in this region to be assessed with regard to the primary prevention of VTE and suggest the need for more correct information about this aspect of venous thromboembolic disease which is not yet sufficiently well known. PMID- 10388459 TI - [Vascular endothelial growth factor. From basic research to clinical application]. AB - There is increasing evidence to support the concept that growth and metastasis of solid tumors, including those of gastrointestinal tract, is facilitated by neoangiogenesis. Vascular Endothelial Growth Factor (VEGF) is one of the most powerful known inducer of endothelial cell growth. Therefore, VEGF is likely to contribute to tumor growth by promoting angiogenesis and stroma formation both directly, through its neovascularization inducing activity, and indirectly, by increasing vascular permeability. In addition, VEGF facilitates tumor diffusion favouring metastatic spread of cancer cells. In view of these implications, it is important to understand the physiopathological role played by this factor. In this review the authors present the accumulating body of data on the biological and functional properties of VEGF, paying special reference to new evidence on its contribution in tumor immune escape, through a marked inhibition of differentiation and activity of the professional antigen presenting cells (APC), namely dendritic cells (DC). As the molecular and cellular events that underlie the functional role of VEGF in tumor angiogenesis and immune suppression become better defined, rational pharmacological and/or gene therapies can be derived in order to treat those neoplasms, such as pancreatic adenocarcinoma, not well amenable to chemo- and radiotherapy or immunotherapy. PMID- 10388460 TI - Tissue factor pathway inhibitor. AB - The role of Tissue Factor (TF, thromboplastin) as the major factor in initiation of the blood coagulation process has been known for more than 100 years. Its importance for the development of clinical thrombosis, be it venous or arterial, and the complexity of the cell biology of TF, have however been increasingly appreciated over the past 15-20 years. Acting as a cofactor for coagulation factor VIIa, it is now clear that TF is able to activate factor IX. Aberrant expression of TF seems to play a major role in the intravascular coagulations disorders linked to endotoxemia, malignancies, immunological diseases and atherosclerosis. Tissue Factor Pathway Inhibitor (TFPI) is the natural direct inhibitor of TF/FVIIa complex. In this study the physiological role, mechanism of action and pharmacological potential of TFPI are discussed. PMID- 10388462 TI - [A new approach to the treatment of fibromyalgia syndrome. The use of Telo Cypro]. AB - BACKGROUND: The therapy of fibromyalgia (SF), with pharmacological and non pharmacological treatments, is not always effective and the benefits obtained can be unsteady or non-lasting. The aim of this study was to evaluate the effect of a pure copper wire sheet ("Telo Cypro") used as bedsheet, on sleep quality as well as spontaneous and provoked pain. METHODS: The study was double-blind, with two parallel groups, versus placebo. Forty patients with fibromyalgia were enrolled, thirty-eight females and two males, with a mean age of 48.8 years and without any current pharmacological treatment. RESULTS: The results obtained show how the use of "Telo Cypro" is extremely beneficial in subjects with fibromyalgia, in reducing painful symptomatology at the tender point level and improving sleep quality, with a positive effect on the patients' cenesthesia at awakening. CONCLUSIONS: In conclusion, the use of "Telo Cypro" can be a valid help in the treatment of fibromyalgia. PMID- 10388461 TI - [Pefloxacin in the treatment of the bone and joint infections]. AB - BACKGROUND: To assess efficacy and tolerability of pefloxacin in association with other antibiotics in the treatment of acute and chronic bone and joint infections. METHODS: From January to December 1997, all the outpatients with diagnosis of acute or chronic bone and joint infections have been enrolled in a perspective study. If possible a cultural or histopathological study was performed. Treatment response was evaluated with radiological and clinical chemistry parameters. RESULTS: Fifteen patients [10 males, 5 females; mean age 40.7 +/- 15 years (range 15-71)] have been studied. They had 5 knee septic arthritis, 1 sacroileitis, 1 hip septic arthritis, 4 long bone osteomyelitis, 1 sterum osteomyelitis, 3 spondilitis. Three patients were HIV infected. Twelve were acute infections, 3 chronic ones. Overall, 7 were hematogenous infections, 6 subsequent to elective surgery, 1 post-traumatic thighbone osteomyelitis, 1 osteomyelitis by external fixation device. Isolates were S. aureus in 5 cases, P. mirabilis in 1 case, S. aureus+ Serratia marcescens in 1. In the remaining part cultural tests were negative. Pefloxacin was administered i.v. or orally at the dose of 400 mg/bid for a mean time of 114 +/- 74.5 days (range 30-270) in association with other chemotherapic agents. Ten good recoveries, 3 partial and 2 no responses were observed. CONCLUSIONS: Pefloxacin resulted to be useful in the treatment of acute and chronic bone and joint infections. No severe side effect was observed during the treatment. PMID- 10388463 TI - [Choristomatous and amartomatous malformations: which is the practical advantage of distinguishing them?]. PMID- 10388464 TI - [What is the role of the electromyographic test exactly?]. PMID- 10388465 TI - Cystic fibrosis syndrome: a new paradigm for inherited disorders and implications for molecular diagnostics. PMID- 10388466 TI - Porphyria conundrum. PMID- 10388467 TI - Prevalence-value-accuracy plots: a new method for comparing diagnostic tests based on misclassification costs. AB - The clinical accuracy of diagnostic tests commonly is assessed by ROC analysis. ROC plots, however, do not directly incorporate the effect of prevalence or the value of the possible test outcomes on test performance, which are two important factors in the practical utility of a diagnostic test. We describe a new graphical method, referred to as a prevalence-value-accuracy (PVA) plot analysis, which includes, in addition to accuracy, the effect of prevalence and the cost of misclassifications (false positives and false negatives) in the comparison of diagnostic test performance. PVA plots are contour plots that display the minimum cost attributable to misclassifications (z-axis) at various optimum decision thresholds over a range of possible values for prevalence (x-axis) and the unit cost ratio (UCR; y-axis), which is an index of the cost of a false-positive vs a false-negative test result. Another index based on the cost of misclassifications can be derived from PVA plots for the quantitative comparison of test performance. Depending on the region of the PVA plot that is used to calculate the misclassification cost index, it can potentially lead to a different interpretation than the ROC area index on the relative value of different tests. A PVA-threshold plot, which is a variation of a PVA plot, is also described for readily identifying the optimum decision threshold at any given prevalence and UCR. In summary, the advantages of PVA plot analysis are the following: (a) it directly incorporates the effect of prevalence and misclassification costs in the analysis of test performance; (b) it yields a quantitative index based on the costs of misclassifications for comparing diagnostic tests; (c) it provides a way to restrict the comparison of diagnostic test performance to a clinically relevant range of prevalence and UCR; and (d) it can be used to directly identify an optimum decision threshold based on prevalence and misclassification costs. PMID- 10388468 TI - Human anti-animal antibody interferences in immunological assays. AB - PURPOSE: The scope and significance of human anti-animal antibody interference in immunological assays is reviewed with an emphasis on human anti-animal immunoglobulins, particularly human anti-mouse antibodies (HAMAs). ISSUES: Anti animal antibodies (IgG, IgA, IgM, IgE class, anti-isotype, and anti-idiotype specificity) arise as a result of iatrogenic and noniatrogenic causes and include human anti-mouse, -rabbit, -goat, -sheep, -cow, -pig, -rat, and -horse antibodies and antibodies with mixed specificity. Circulating antibodies can reach gram per liter concentrations and may persist for years. Prevalence estimates for anti animal antibodies in the general population vary widely and range from <1% to 80%. Human anti-animal antibodies cause interferences in immunological assays. The most common human anti-animal antibody interferent is HAMA, which causes both positive and negative interferences in two-site mouse monoclonal antibody-based assays. Strategies to prevent the development of human anti-animal antibody responses include immunosuppressant therapy and the use of humanized, polyethylene glycolylated, or Fab fragments of antibody agents. Sample pretreatment or assay redesign can eliminate immunoassay interferences caused by anti-animal antibodies. Enzyme immunoassays, immunoradiometric assays, immunofluorescence, and HPLC assays have been designed to detect HAMA and other anti-animal antibodies, but intermethod comparability is complicated by differences in assay specificity and lack of standardization. CONCLUSIONS: Human anti-animal antibodies often go unnoticed, to the detriment of patient care. A heightened awareness on the part of laboratory staff and clinicians of the problems caused by this type of interference in routine immunoassay tests is desirable. Efforts should be directed at improving methods for identifying and eliminating this type of analytical interference. PMID- 10388469 TI - Detection of five rare cystic fibrosis mutations peculiar to Southern Italy: implications in screening for the disease and phenotype characterization for patients with homozygote mutations. AB - BACKGROUND: The search for the eight most frequent mutations (i.e., DeltaF508, G542X, W1282X, N1303K, 1717-1G-->A, R553X, 2183AA-->G, and I148T) by allele specific oligonucleotide dot-blot analysis revealed 78% of 396 cystic fibrosis alleles in Southern Italy. The observation of frequent haplotypes on the unidentified cystic fibrosis alleles suggested that a few mutations could account for a large number of unidentified alleles. METHODS: We screened most of the coding sequence of the cystic fibrosis transmembrane regulator gene by denaturing gradient gel electrophoresis to determine the spectrum of these mutations in 68 unrelated cystic fibrosis patients bearing one or both unidentified mutations. RESULTS: The screening revealed five mutations, R1158X, 711+1G-->T, 4016insT, L1065P, and G1244E, each of which had a frequency of 1.3-1.8% (7% collectively). The 7% increase in the detection rate (85% vs 78%) reduces by >50% the residual risk of being cystic fibrosis carriers for couples who had tested negative by molecular analysis. We therefore designed a second allele-specific oligonucleotide set to analyze the five mutations. Among the patients analyzed, one patient homozygous for the L1065P mutation expressed a mild pulmonary and intestinal form of the disease with pancreatic insufficiency. Two other patients, homozygous for mutations R1158X and 4016insT, both expressed a severe cystic fibrosis phenotype. CONCLUSIONS: Five cystic fibrosis mutations are peculiar to patients from Southern Italy. The method described for their analysis is efficient, inexpensive, and can be semi-automated by use of a robotic workstation. The results obtained in patients from Southern Italy may have an impact on laboratories in other countries, given the large migrations of populations from Southern Italy to other countries in the last two centuries. PMID- 10388470 TI - The lipoprotein lipase HindIII polymorphism: association with total cholesterol and LDL-cholesterol, but not with HDL and triglycerides in 342 females. AB - BACKGROUND: Lipoprotein lipase (LPL) is the rate-limiting enzyme in the hydrolysis of core triglycerides in chylomicrons and VLDL. METHODS: We investigated the association between the HindIII polymorphism of the LPL gene and fasting glucose, lipid, and lipoprotein concentrations in 683 Caucasians. We first stabilized the study subjects, using an 8-day diet and exercise intervention program before obtaining blood samples. The use of this standardization period reduced the variance of all glucose and lipid concentrations. RESULTS: In our study, the HindIII allele frequencies for females and males were 0.29 and 0.34 for H- and 0.71 and 0.66 for H+, respectively. We found in females, but not in males, a significant association between the HindIII genotype and total cholesterol (P = 0.007) and LDL-cholesterol (P = 0.018), with females homozygous for the rare H- allele having the lowest, heterozygotes (H-/+) having intermediate, and women homozygous for the common H+ allele having the highest of each of these lipid traits. With regard to triglycerides, HDL cholesterol, and glucose, no significant effect of the HindIII genotype was noted in either gender. CONCLUSIONS: These results suggest that in a gender-specific manner, the rare LPL HindIII H- allele has a cholesterol-lowering and, therefore, potentially cardioprotective effect compared with the common H+ allele. PMID- 10388471 TI - Evaluation of cation-exchange HPLC compared with isoelectric focusing for neonatal hemoglobinopathy screening. AB - BACKGROUND: Central Middlesex Hospital, in northwest London, has screened neonates for hemoglobinopathies, using the established manual technique of isoelectric focusing (IEF) since 1989. Recently, this laboratory has faced a large increase in the number of samples tested per year. This study compared the detection of hemoglobin abnormalities between the existing manual IEF method and that of automated cation-exchange HPLC to determine the reliability of HPLC and whether an automated system would save time in the laboratory. METHODS: Over a 15 month period, 25 750 blood samples, collected by heel prick onto filter paper, were tested using HPLC, and the results were compared with those obtained with IEF. RESULTS: HPLC and IEF each identified 568 patients with FAS, 151 with FAC, 49 with FAD-Punjab, 23 with FS, 3 with FC, 6 with FSC, 5 with FE, and 1 with FD. IEF detected 62 patients with FAE, whereas HPLC detected 63. This additional FAE was observed on repeat IEF. One additional heterozygote detected by HPLC was initially not observed by IEF, but was detected on repeat IEF. HPLC detected all but six cases of Hb Barts observed by IEF. One double heterozygote and four heterozygotes were detected by IEF, but not by HPLC. The detection of hemoglobin variants expressed at low concentrations was comparable for the two methods, and carryover was not observed in routine analysis on HPLC. CONCLUSIONS: HPLC is a sensitive, efficient, and time-saving alternative to IEF for the neonatal screening of common hemoglobinopathies. PMID- 10388472 TI - BRCA1 gene mutations in sporadic ovarian carcinomas: detection by PCR and reverse allele-specific oligonucleotide hybridization. AB - BACKGROUND: Although germline mutations in BRCA1 play a central role in familial breast and ovarian cancers, to date, no somatic mutations in BRCA1 have been reported in sporadic breast cancer, and only five somatic mutations have been identified in the sporadic ovarian carcinomas. Because loss of heterozygosity appears frequently at the BRCA1 locus in nonfamilial breast and ovarian carcinomas, we searched for mutations in the BRCA1 gene in sporadic ovarian tumors. METHODS: We developed a detection system based on PCR and reverse allele specific oligonucleotide hybridization on membrane strips for the simultaneous detection of 17 frequently occurring mutations in the BRCA1 gene. RESULTS: As little as 2% mutant DNA in a sample could be detected. Two of 122 DNA samples isolated from sporadic ovarian tumor biopsies contained the Cys61Gly mutation. Both mutations were germline mutations. One of these was an ovarian metastasis of a primary fallopian tube carcinoma. The tubal carcinoma was also confirmed to contain the Cys61Gly mutation. CONCLUSIONS: This is the first report that a germline BRCA1 mutation is associated with primary tubal carcinoma. The 17 specific mutations in the BRCA1 gene do not play a major role in the tumorigenesis and progression of sporadic ovarian cancer. PMID- 10388473 TI - TaqMan PCR-based gene dosage assay for predictive testing in individuals from a cancer family with INK4 locus haploinsufficiency. AB - BACKGROUND: A genetic syndrome of cutaneous malignant melanoma and nervous system tumors recently has been characterized and shown to be linked to the INK4 locus in the 9p21 region. Hemizygosity at adjacent physically mapped microsatellite markers indicated deletion of p16, p19, and p15 clustered tumor suppressors. Because individuals from this family could benefit from predictive testing in terms of cancer prevention, we developed a direct test without need to analyze parental DNAs to comply with the rules of individual consent and secrecy. METHODS: We developed an assay using TaqManTM real-time quantitative PCR, with p15 as the test sequence and albumin (ALB) as the reference gene. The normalized ratio of p15/ALB is expected to yield a value of approximately 1 in individuals without the deletion, whereas a ratio of approximately 0.5, indicating p15 haploinsufficiency, is expected in predisposed individuals. RESULTS: All patients harboring the previously defined at-risk haplotype were correctly identified using this approach. In six individuals with deletions, the p15/ALB ratios were 0.472-0.556 (SD, 0.013-0.078). In the five individuals without deletions, the ratios were 0.919-1.019 (SD, 0.006-0.075). CONCLUSIONS: This is the first report of a high-throughput, automatable gene dosage assay successfully applied to the identification of a germ-line deletion. This approach, not limited by marker informativeness or the need for harvesting live cells, can be applied to any condition with haploinsufficiency and extended to the characterization of most abnormalities of the ploidy. PMID- 10388474 TI - Estimation of prostate cancer probability by logistic regression: free and total prostate-specific antigen, digital rectal examination, and heredity are significant variables. AB - BACKGROUND: Despite low specificity, serum prostate-specific antigen (PSA) is widely used in screening for prostate cancer. Specificity can be improved by measuring free and total PSA and by combining these results with clinical findings. Methods such as neural networks and logistic regression are alternatives to multistep algorithms for clinical use of the combined findings. METHODS: We compared multilayer perceptron (MLP) and logistic regression (LR) analysis for predicting prostate cancer in a screening population of 974 men, ages 55-66 years. The study sample comprised men with PSA values >3 microg/L. Explanatory variables considered were age, free and total PSA and their ratio, digital rectal examination (DRE), transrectal ultrasonography, and a family history of prostate cancer. RESULTS: When at least 90% sensitivity in the training sets was required, the mean sensitivity and specificity obtained were 87% and 41% with LR and 85% and 26% with MLP, respectively. The cancer specificity of an LR model comprising the proportion of free to total PSA, DRE, and heredity as explanatory variables was significantly better than that of total PSA and the proportion of free to total PSA (P <0.01, McNemar test). The proportion of free to total PSA, DRE, and heredity were used to prepare cancer probability curves. CONCLUSION: The probability calculated by logistic regression provides better diagnostic accuracy for prostate cancer than the presently used multistep algorithms for estimation of the need to perform biopsy. PMID- 10388476 TI - Quantitative bedside assay for cardiac troponin T: a complementary method to centralized laboratory testing. AB - BACKGROUND: In the evaluation of chest pain patients, whole blood bedside assays of highly specific cardiac molecules may be an attractive alternative to centralized clinical chemistry testing. We now report on an optimized test strip device for cardiac troponin T (cTnT) that can be analyzed by a cardiac reader for quantitative assessment of the test result. METHODS AND RESULTS: The cTnT test strip reader measures, via a CCD camera, the reflectance of the signal line. For quantitative analysis, a calibration curve was constructed from 1030 samples of 252 consecutive patients with acute coronary syndromes. In a method comparison of 140 samples, the quantitative results of the cTnT test strip reader correlated closely with the results of the cTnT ELISA (r = 0.98; y = 0.85x + 0. 002). Within run and day-to-day (n = 10) mean CVs were between 11% and 16%, respectively. The cross-reactivity with skeletal troponin T was <0.02%. In patients with myocardial infarction, 45% and 91% of all samples were positive on admission and at 4-8 h after the onset of symptoms, respectively. ROC curve analysis demonstrated a comparable efficiency of the cTnT test strip reader and the laboratory-based cTnT ELISA in patients with suspected myocardial infarction. CONCLUSIONS: It is now possible to quantitatively determine cTnT at the patient's bedside with a rapid and convenient test device. This will facilitate the diagnostic work up of patients with suspected myocardial cell necrosis. PMID- 10388475 TI - Optimization of allopurinol challenge: sample purification, protein intake control, and the use of orotidine response as a discriminative variable improve performance of the test for diagnosing ornithine carbamoyltransferase deficiency. AB - BACKGROUND: The diagnosis of heterozygosity for X-linked ornithine carbamoyltransferase (OCT) deficiency has usually been based on measurement of the increase of orotate and orotidine excretion after an allopurinol load. We examined the choices of analyte, cutoff, and test conditions to obtain maximal test accuracy. METHODS: Urine orotate/orotidine responses to allopurinol load in 37 children (13 OCT-deficient and 24 non-OCT-deficient) and 24 women (7 at risk for carrier status and 17 not related to OCT-deficient children) were analyzed by liquid chromatography after sample purification by anion-exchange chromatography. Diagnostic accuracy was evaluated by nonparametric ROC curves. RESULTS: Sample purification was necessary to prevent interferences. Orotate and orotidine excretion increased with increased protein intake during the test. At a cutoff of 8 mmol orotidine/mol creatinine, sensitivity was 1.0 and specificity was 0. 92 in mild forms of OCT deficiency. Results in monoplex carrier women may differ greatly from those expected because of the genetics of this deficiency. CONCLUSIONS: Standardization of protein intake is required in the allopurinol loading test. A negative response in the face of clinical suspicion should be followed with a repeat test during a protein intake not <2.5 g x kg-1 x day-1. Measurements of orotidine provide better clinical sensitivity than measurements of orotate. PMID- 10388477 TI - Use of bone alkaline phosphatase to monitor alendronate therapy in individual postmenopausal osteoporotic women. AB - BACKGROUND: Biochemical bone markers are sensitive to the changes in bone turnover that result from treatment of postmenopausal osteoporotic women with antiresorptive therapies. Although information is available on the use of bone markers in monitoring therapy in groups of subjects, less is known regarding how these markers perform in individual patients. METHODS: Serum bone alkaline phosphatase (bone ALP) concentrations, measured with the Tandem(R) Ostase(R) assay, were used to monitor the biochemical response of bone in postmenopausal women with osteoporosis receiving either 10 mg/day alendronate therapy (n = 74) or calcium supplementation (n = 148) for 24 months. RESULTS: Bone ALP decreased significantly from baseline at 3 months (P Trp) associated with two new haplotypes and evidence that apo B-100 (Glu3405-->Gln) diminishes receptor-mediated uptake of LDL. AB - BACKGROUND: Ligand-defective apolipoprotein (apo) B-100 is a major cause of hypercholesterolemia. For many years, apo B-100 (Arg3500-->Gln) has been the only mutation known to cause ligand-defective apo B-100. METHODS: Using temperature gradient gel electrophoresis, we screened 297 unrelated individuals with LDL cholesterol >1.55 g/L and triglycerides <2.0 g/L for sequence variants of the putative LDL receptor-binding domain of apo B-100. RESULTS: We found apo B-100 (Arg3500-->Gln) in 21 individuals (7.1%). When extrapolated to the general population, this corresponds to the highest prevalence of apo B-100 (Arg3500- >Gln) reported to date. Furthermore, we identified three unrelated carriers (1%) of a silent substitution (CTG-->CTA) affecting the codon for leucine3350, four carriers (1.3%) of apo B-100 (Glu3405-->Gln), and two subjects (0.7%) with apo B 100 (Arg3500-->Trp). apo B-100 (Arg3500-->Trp) was assigned to two different, previously unknown haplotypes. The binding, uptake, and degradation of apo B-100 (Arg3500-->Trp) was lower than that of normal LDL, but higher than with apo B-100 (Arg3500-->Gln), implying that the substitution of Trp3500 for Arg may cause less severe reduction of binding than the substitution of Gln. LDL from individuals heterozygous for apo B-100 (Glu3405-->Gln) bound to LDL receptors at the same rate as normal LDL, but was taken up and degraded at significantly reduced rates, suggesting that domains of apo B-100 involved in binding and uptake do not completely overlap. CONCLUSIONS: In Germany, apo B-100 (Arg3500-->Gln) may be more frequent than previously assumed. Both apo B-100 (Arg3500-->Trp) and apo B 100 (Glu3405-->Gln) may contribute to the phenotype of ligand-defective LDL. These variants will be missed if screening is confined to apo B-100 (Arg3500- >Gln) only. PMID- 10388480 TI - Lipoprotein(a)-cholesterol and coronary heart disease in the Framingham Heart Study. AB - BACKGROUND: Increased plasma lipoprotein(a) [Lp(a)] concentrations have been reported to be an independent risk factor for coronary heart disease (CHD) in some prospective studies, but not in others. These inconsistencies may relate to a lack of standardization and the failure of some immunoassays to measure all apolipoprotein(a) isoforms equally. METHODS: We measured plasma Lp(a)-cholesterol [Lp(a)-C] in a Caucasian population of offspring and spouses of the Framingham Heart Study participants, using a lectin-based assay (LipoproTM). We compared the prevalence of increased Lp(a)-C to the presence of sinking pre-beta-lipoprotein (SPB). We also related Lp(a)-C concentrations to the prevalence of CHD risk in the entire population. RESULTS: The mean (+/- SD) Lp(a)-C concentration in the Framingham population (n = 3121) was 0.186 +/- 0.160 mmol/L, with no significant gender or age differences. The mean Lp(a)-C concentrations in the absence or presence of SPB were 0.158 +/- 0. 132 mmol/L and 0.453 +/- 0.220 mmol/L, respectively (P <0.0001). The mean Lp(a)-C concentration in men with CHD (n = 156) was 0.241 +/- 0. 204 mmol/L, which was significantly (P <0.001) higher, by 34%, than in controls. The odds ratio for CHD risk in men with Lp(a)-C >/=0. 259 mmol/L (>/=10 mg/dL), after adjusting for age, HDL-cholesterol, LDL-cholesterol, smoking, diabetes, blood pressure, and body mass index, was 2.293 (confidence interval, 1.55-3.94; P <0.0005). Lp(a)-C values correlated highly with a Lp(a) mass immunoassay [ApotekTM Lp(a); r = 0.832; P <0.0001; n = 1000]. CONCLUSIONS: An increased Lp(a)-C value >/=0.259 mmol/L (>/=10 mg/dL) is an independent CHD risk factor in men with a relative risk of more than 2, but was inconclusive in women. Lp(a)-C measurements offer an alternative to Lp(a)-mass immunoassays and can be performed on automated analyzers. PMID- 10388481 TI - The apparent inhibition of inosine monophosphate dehydrogenase by mycophenolic acid glucuronide is attributable to the presence of trace quantities of mycophenolic acid. AB - BACKGROUND: Mycophenolic acid glucuronide, the primary metabolite of the immunosuppressive agent mycophenolic acid, affords weak inhibition of proliferating and resting lymphocytes and recombinant human inosine monophosphate dehydrogenase in comparison to the active drug. We evaluated the hypothesis that mycophenolic acid is a trace contaminant of the glucuronide metabolite preparation and that this accounts for the observed effects of mycophenolic acid glucuronide on human inosine monophosphate dehydrogenase catalytic activity both in lymphocytes and the pure enzyme. METHODS: We used negative ion electrospray HPLC-mass spectrometry (HPLC-MS) and HPLC-tandem MS (HPLC-MS-MS) to identify mycophenolic acid as a contaminant of mycophenolic acid glucuronide. Quantification of the mycophenolic acid contaminant was achieved using a negative ion electrospray HPLC-MS method in the selected-ion monitoring mode. RESULTS: Trace amounts of mycophenolic acid were detected and definitively identified in the mycophenolic acid glucuronide preparation by the HPLC-MS-MS analysis. In addition to having identical HPLC retention times, pure mycophenolic acid and the contaminant produced the following major fragments upon HPLC-MS-MS analysis: deprotonated molecular ion, m/z 319; and fragment ions, m/z 275, 243, 205, and 191 (the most abundant fragment ion). Using the negative ion electrospray HPLC-MS procedure in the selected-ion monitoring mode, the quantity of the contaminant mycophenolic acid was determined to be 0.312% +/- 0.0184% on a molar basis. CONCLUSION: These data provide strong support for the proposal that the apparent inhibition of the target enzyme inosine monophosphate dehydrogenase by mycophenolic acid glucuronide is attributable to the presence of trace amounts of contaminant mycophenolic acid. PMID- 10388482 TI - Adulteration of urine by "Urine Luck". AB - BACKGROUND: In vitro adulterants are used to invalidate assays for urine drugs of abuse. The present study examined the effect of pyridinium chlorochromate (PCC) found in the product "Urine Luck". METHODS: PCC was prepared and added to positive urine controls at concentrations of 0, 10, 50, and 100 g/L. The controls were assayed for methamphetamine, benzoylecgonine (BE), codeine and morphine, tetrahydrocannabinol (THC), and phencyclidine (PCP) with the Emit II (Syva) and Abuscreen Online (Roche) immunoassays, and by gas chromatography/mass spectrometry (GC/MS). Two tests were also developed to detect PCC in urine: a spot test to detect chromate ions using 10 g/L 1,5-diphenylcarbazide as the indicator, and a GC/MS assay for pyridine. We tested 150 samples submitted for routine urinalysis, compliance, and workplace drug testing for PCC, using these assays. RESULTS: Response rates decreased at 100 g/L PCC for all Emit II drug assays and for the Abuscreen morphine and THC assays. In contrast, the Abuscreen amphetamine assay produced apparently higher results, and no effect was seen on the results for BE or PCP. The PCC did not affect the GC/MS recovery of methamphetamine, BE, PCP, or their deuterated internal standards, but decreased GC/MS recovery of the opiates at both intermediate (50 g/L) and high (100 g/L) PCC concentrations and apparent concentrations of THC and THC-d3 at all PCC concentrations. Two of 50 samples submitted for workplace drug testing under chain-of-custody conditions were positive for PCC, whereas none of the remaining 100 specimens submitted for routine urinalysis or compliance drug testing were positive. CONCLUSIONS: PCC is an effective adulterant for urine drug testing of THC and opiates. Identification of PCC use can be accomplished with use of a spot test for the oxidant. PMID- 10388483 TI - Stereospecific analysis and enantiomeric disposition of 3, 4 methylenedioxymethamphetamine (Ecstasy) in humans. AB - BACKGROUND: Little is known concerning the enantioselective disposition of 3,4 methylenedioxymethamphetamine (MDMA; ecstasy) in humans. In addition, the potential of utilizing the stereochemical composition of an analyte in biological media for forensic purposes requires investigation. METHODS: The enantiomers of MDMA and its demethylated metabolite, 3,4-methylenedioxyamphetamine (MDA), present in plasma and urine extracts were derivatized with (-)-(R)-alpha-methoxy alpha-trifluoromethylphenylacetyl chloride and analyzed by gas chromatography mass spectrometry and gas chromatography, respectively. The enantioselective disposition of MDMA and MDA was determined following oral administration of racemic MDMA (40 mg) to eight male volunteers. RESULTS: The plasma concentrations of (R)-MDMA exceeded those of the S-enantiomer [ratio R:S of the area under the curve (AUC), 2.4 +/- 0.3], and the plasma half-life of (R)-MDMA (5.8 +/- 2.2 h) was significantly longer than that of the S-enantiomer (3.6 +/- 0.9 h). The majority of the recovered material in urine was excreted within 24 h after dosing, with the recovery of (R)-MDMA (21.4% +/- 11.6%) being significantly greater than that of (S)-MDMA (9.3% +/- 4.9%), and with (S)- and (R)-MDA accounting for 1.4% +/- 0.5% and 1.0% +/- 0.3% of the dose, respectively. Mathematical modeling of plasma enantiomeric composition vs sampling time demonstrated the applicability of using stereochemical data for the prediction of time elapsed after drug administration. CONCLUSIONS: Analytical methods for determining the enantiomeric composition of MDMA and MDA in plasma and urine were developed. The disposition of MDMA in humans is stereoselective, with the more active S-enantiomer having a reduced AUC and shorter half-life than (R)-MDMA. The determination of stereochemical composition may be applicable for forensic purposes. PMID- 10388484 TI - Plasma porphyrins in the porphyrias. AB - BACKGROUND: As an aid in the diagnosis and management of porphyria we have developed a method to fractionate and quantify plasma porphyrins and have evaluated its use in various porphyrias. METHODS: We used HPLC with fluorometric detection to measure plasma concentrations of uroporphyrin I and III, heptacarboxyl III, hexacarboxyl III, pentacarboxyl III, and coproporphyrin I and III. We studied 245 healthy subjects, 32 patients with classical porphyria cutanea tarda (PCT), 12 patients with PCT of renal failure, 13 patients with renal failure, 8 patients with pseudoporphyria of renal failure, 3 patients with acute intermittent porphyria, 5 patients with variegate porphyria, 5 patients with hereditary coproporphyria, and 4 patients with erythropoietic protoporphyria. RESULTS: Between-run CVs were 5.4-13%. The recoveries of porphyrins added to plasma were 71-114% except for protoporphyrin, which could not be reliably measured with this technique. Plasma porphyrin patterns clearly identified PCT, and its clinical sensitivity equaled that of urine porphyrin fractionation. The patterns also allowed differentiation of PCT of renal failure from pseudoporphyria of renal failure. CONCLUSIONS: The assay of plasma porphyrins identifies patients with PCT and appears particularly useful for differentiating PCT of renal failure from pseudoporphyria of renal failure. PMID- 10388485 TI - The use of infrared spectrophotometry for measuring body water spaces. AB - BACKGROUND: The conventional method of measuring total body water by the deuterium isotope dilution method uses gas isotope ratio mass spectrometry (IRMS), which is both expensive and time-consuming. We investigated an alternative method, using Fourier transform infrared spectrophotometry (FTIR), which uses less expensive instrumentation and requires little sample preparation. METHOD: Total body water measurements in human subjects were made by obtaining plasma, saliva, and urine samples before and after oral dosing with 1.5 mol of deuterium oxide. The enrichments of the body fluids were determined from the FTIR spectra in the range 1800-2800 cm-1, using a novel algorithm for estimation of instrumental response, and by IRMS for comparison. RESULTS: The CV (n = 5) for repeat determinations of deuterium oxide in biological fluids and calibrator solutions (400-1000 micromol/mol) was found to be in the range 0.1-0.9%. The use of the novel algorithm instead of the integration routines supplied with the instrument gave at least a threefold increase in precision, and there was no significant difference between the results obtained with FTIR and those obtained with IRMS. CONCLUSION: This improved infrared method for measuring deuterium enrichment in plasma and saliva requires no sample preparation, is rapid, and has potential value to the clinician. PMID- 10388486 TI - Automated albumin method underestimates pharmaceutical-grade albumin in vivo. PMID- 10388487 TI - Thiols as a measure of plasma redox status in healthy subjects and in patients with renal or liver failure. PMID- 10388488 TI - Pediatric reference intervals for serum thyroxine, triiodothyronine, thyrotropin, and free thyroxine. PMID- 10388489 TI - Serum potassium is unreliable as an estimate of in vivo plasma potassium. PMID- 10388490 TI - Genotyping method for point mutation detection in the intestinal fatty acid binding protein, using fluorescent probes. PMID- 10388491 TI - High-speed apolipoprotein E genotyping and apolipoprotein B3500 mutation detection using real-time fluorescence PCR and melting curves. PMID- 10388492 TI - Biological day-to-day variation and reference change limits of serum cortisol and aldosterone in healthy young men on unrestricted diets. PMID- 10388493 TI - LUC, what is that? Large unstained cells. PMID- 10388494 TI - Serum gamma-glutamyltransferase isoform complexed to LDL in the diagnosis of small hepatocellular carcinoma. PMID- 10388495 TI - Prostate-specific antigen expression in normal human bone marrow cells. PMID- 10388497 TI - Compiled by david E. Bruns, editor (dbruns@aacc.org) PMID- 10388496 TI - National Academy of Clinical Biochemistry Standards of Laboratory Practice: recommendations for the use of cardiac markers in coronary artery diseases. AB - The Sixth Conference on the "Standards of Laboratory Practice Series", sponsored by the National Academy of Clinical Biochemistry (NACB), was held on August 4-5, 1998, at the Annual Meeting of the American Association for Clinical Chemistry, in Chicago, IL. An expert committee was assembled to write recommendations on the use of cardiac markers in coronary artery diseases. The NACB Committee prepared a preliminary draft of the guidelines, made them available on the World Wide Web (www.nacb.org), and distributed them before the presentations. The recommendations were divided into four areas: the use of markers in the triage of patients with chest pain, acute coronary syndromes, clinical applications other than acute myocardial infarction and research, and assay platforms and markers of acute myocardial infarction. The recommendations were revised and subsequently re presented in part at the "Biomarkers in Acute Cardiac Syndromes Conference", sponsored by the Jewish Hospital Heart and Lung Institute, Louisville KY, on October 16-17, 1998. This report lists each recommendation, its scientific justification, and a summary of discussions from conference participants and reviewers. Approximately 100 individuals responded to various versions of these recommendations via direct correspondences, telephone calls to Committee members, electronic mail correspondence to the Committee Chairman, or oral questions and comments raised during one of the two conference presentations. Some of the recommendations were changed to reflect the consensus opinion. In cases in which there was no consensus, the Committee included pertinent discussion without necessarily changing the original recommendations. At times, the Committee members felt that although a particular recommendation might not be the current standard of care today, they anticipate that it likely will be adopted in the near future. PMID- 10388498 TI - Banff schema for grading liver allograft rejection: utility in clinical practice. AB - An accurate and functional system for grading acute liver allograft rejection is important for patient management, research, and communication. The Banff schema is a consensus document designed to provide an internationally accepted standard for this purpose. The aim of this study is to determine if application of the Banff schema would significantly alter the grading of acute liver allograft rejection compared with the Birmingham system. One hundred twenty-four post-liver transplantation biopsies performed by the Western Australian Liver Transplantation Service between 1992 and 1997 were retrospectively analyzed by a pathologist and a hepatologist. Each was supplied with a brief clinical history before applying the Banff and Birmingham criteria. Results were compared with each other and to the diagnosis made at the time of the biopsy, which was based on the European grading system. Rejection was diagnosed by the reviewers in 61 of 124 biopsy specimens according to the criteria of Snover. The Banff schema and Birmingham system agreed on the grade of rejection in 22 of the 61 biopsy specimens. The Banff schema elevated the grade of rejection in 39 specimens by an increment of one. In no instance did the Banff schema reduce the grade. Comparison between the Banff schema and diagnosis made at the time of biopsy showed agreement in 39 specimens, whereas the Banff schema elevated the grade in 15 specimens and reduced the grade in 23 specimens. In comparison to the Birmingham system, the Banff schema elevated the grade of liver allograft rejection in the majority of biopsy specimens, and this has the potential to alter clinical management with the adoption of the Banff schema or if the systems are used interchangeably. PMID- 10388500 TI - Endoscopic stenting of the gallbladder for symptomatic gallbladder disease in patients with end-stage liver disease awaiting orthotopic liver transplantation. AB - Cholecystectomy in patients with advanced cirrhosis is associated with excessive morbidity and mortality. Because open cholecystectomy in patients with Child's class C cirrhosis has a reported mortality rate as high as 83%, symptomatic gallbladder disease in patients awaiting orthotopic liver transplantation (OLT) poses a unique clinical problem. The goal of this study is to determine whether the treatment of symptomatic gallbladder disease with endoscopic stenting of the gallbladder effectively reduces biliary symptoms and complications or the need for cholecystectomy. Thirteen patients with symptomatic gallbladder disease with and without cholelithiasis and advanced cirrhosis who were candidates for OLT underwent placement of a biliary stent from the gallbladder to the duodenum at endoscopic retrograde cholangiography. In each patient, biliary symptoms and complications ceased after stent placement. Seven patients underwent successful OLT 1 to 24 months after the procedure. One patient subsequently became a noncandidate for OLT and died of diabetes complications 3 years after the procedure. Five others are awaiting OLT (6 to 28 months postprocedure). One patient had recurrent pericholecystic fluid collection requiring percutaneous drainage and antibiotic therapy 8 months after the procedure. No patient has had recurrent symptoms, and currently all patients are free of complications. None required surgical intervention of the gallbladder or biliary tree. We conclude that endoscopic stenting of the gallbladder is the preferred treatment for symptomatic gallbladder disease in patients with end-stage liver disease awaiting OLT. This approach is noninvasive, safe, and effective in preventing potential morbidity and mortality. PMID- 10388499 TI - Effect of pretransplantation ursodeoxycholic acid therapy on the outcome of liver transplantation in patients with primary biliary cirrhosis. AB - As ursodeoxycholic acid (UDCA) delays the need for transplantation, this could result in patients with more comorbid disease, therefore more likely to have a worse outcome posttransplantation. The aim of this study is to compare posttransplantation outcome in patients who received UDCA versus placebo who subsequently required a liver transplant. Data on all trial patients referred for transplantation were retrospectively collected from three randomized controlled trials of UDCA in patients with primary biliary cirrhosis (PBC). An intent-to treat analysis was conducted with patients assigned to their original treatment allocation. UDCA and placebo groups were compared at trial entry, transplant referral, just before transplantation, and 1 month and 1 year posttransplantation. From 1987 to 1996, 37 UDCA-treated and 53 placebo patients were referred for transplantation; their age, sex, and serum bilirubin levels were similar at study entry. Immediately before transplantation, these two groups were again similar with respect to age, bilirubin level, Mayo risk score, and serum creatinine level. Posttransplantation survival rates at 1 month were 93.9% in the UDCA group and 88.4% in the placebo group, and 1 year survival rates were 90.3% and 78.4%, respectively (not significant). Posttransplantation, the two groups had similar rates of infection (53.9% v 58%); however, rejection occurred significantly less often in the UDCA group; 42.9% versus 68. 8% in the placebo group (P =.04). The posttransplantation outcome of UDCA-treated patients with PBC is no different from those who were administered placebo. There is no evidence to suggest the beneficial effect of UDCA in delaying the need for transplantation is associated with a worse outcome once liver transplantation becomes necessary. PMID- 10388501 TI - Activation of nuclear factor-kappaB during orthotopic liver transplantation in rats is protective and does not require Kupffer cells. AB - Reperfusion after liver transplantation results in the induction of tumor necrosis factor-alpha (TNFalpha) as well as activation of the stress-associated signaling proteins, c-Jun N-terminal kinase (JNK), activating protein-1 (AP-1), and nuclear factor-kappaB (NF-kappaB). To test the hypothesis that Kupffer cells are involved in the activation of signal transduction cascades during rat liver transplantation, Kupffer cells were depleted from donor liver using gadolinium chloride (GdCl3), and then the activation of JNK, AP-1, and NF-kappaB were assessed after transplantation. The results showed that GdCl3 treatment did not inhibit the activation of these stress signals, although transplanted livers were depleted of Kupffer cells and partially protected from reperfusion injury. Interleukin-6 (IL-6) and IL-10 messenger RNAs (mRNAs) were induced by transplantation, and the induction was suppressed by Kupffer cell depletion. The induction of TNFalpha mRNA and serum protein during liver transplantation was unaffected by GdCl3. These results show that Kupffer cells are not a major source of TNFalpha production after liver transplantation and that stress-signaling protein activation occurs independently of Kupffer cells. Transplantation strongly activates the transcription factor NF-kappaB, which blocks TNFalpha mediated apoptosis in hepatocytes in vitro. To assess the role of NF-kappaB activation during liver transplantation, the IkappaBalpha superrepressor was expressed in donor livers using adenoviral-mediated gene transfer. Inhibition of NF-kappaB resulted in increased serum alanine aminotransferase levels after 3 hours of transplantation. In addition, the blockade of NF-kappaB resulted in increased histological tissue injury and increased hepatic terminal deoxyribonucleotide transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) staining, indicating apoptosis. These results show that NF kappaB activation has a protective role in the transplanted liver. PMID- 10388502 TI - Fibrosing cholestatic hepatitis in renal transplant recipients with hepatitis C virus infection. AB - Fibrosing cholestatic hepatitis (FCH) has been described as a specific manifestation of hepatitis B virus (HBV) infection in liver allograft recipients characterized by a rapid progression to liver failure. Only sporadic cases have been reported in other immunocompromised groups infected with HBV and in a few transplant recipients with hepatitis C virus (HCV) infection. We present the occurrence of FCH in 4 HCV-infected renal transplant recipients within a series of 73 renal transplant recipients with HCV infection followed up closely serologically and with consecutive liver biopsies. All 4 patients received the triple-immunosuppressive regimen (azathioprine, cyclosporine A, methylprednisolone). The interval from transplantation to the appearance of liver dysfunction was 1 to 4 months and to histological diagnosis, 3 to 11 months. The biochemical profile was analogous to a progressive cholestatic syndrome in 3 patients, whereas the fourth patient had only slightly increased alanine aminotransferase and gamma-glutamyl transferase (gammaGT) levels. Liver histological examination showed the characteristic pattern of FCH in 2 patients, whereas the other 2 patients had changes compatible with an early stage. All patients were anti-HCV negative at the time of transplantation, whereas 2 patients, 1 with incomplete and 1with complete histological FCH features, seroconverted after 3 and 31 months, respectively. The patients were HCV RNA positive at the time of the first liver biopsy and showed high serum HCV RNA levels (14 to 58 x 10(6) Eq/mL, branched DNA). HCV genotype was 1b in 3 patients and 3a in 1 patient. After histological diagnosis, immunosuppression was drastically reduced. Two patients died of sepsis and liver failure 16 and 18 months posttransplantation, whereas the seroconverted patients showed marked improvement of their liver disease, which was histologically verified in 1 patient. In conclusion, FCH can occur in HCV-infected renal transplant recipients. It seems to develop as a complication of a recent HCV infection during the period of maximal immunosuppression and is associated with high HCV viremia levels. There are indications that drastic reduction of immunosuppression may have a beneficial effect on the outcome of the disease. PMID- 10388503 TI - Improved physical performance after orthotopic liver transplantation. AB - Orthotopic liver transplantation (OLT) has become a frequently used treatment for end-stage liver disease and acute liver failure, and liver function is markedly improved after transplantation. However, no studies have investigated the development in physical capacity after OLT. On this basis, the aim of the present study is to study the influence of OLT on physical fitness during the first postoperative year. Twenty-three men with a mean age of 45.1 years (range, 24 to 62 years) and 15 women with a mean age of 44.6 years (range, 21 to 62 years) were included in the study. Preoperative maximal oxygen uptake (VO2max) during graded ergometer bicycling, isokinetic knee extension/flexion moments, and functional performance (i.e., 6-minute walking distance and standardized transfers and squats) was measured. Preoperative fitness and strength was 40% to 50% less than expected in the age-matched general population. Post-OLT, all patients underwent a supervised exercise program for 8 to 24 weeks. Follow-up data showed a significant increase in all tested physical performance parameters after OLT. Six months post-OLT, VO2max had increased 43%; knee strength, 60% to 100%; and functional performance, 22% to 27%. One year postsurgery, general health was improved and perceived as excellent or good in all patients. All patients were independent in activities of daily living, and the level of physical activity increased after OLT. No further improvement in either physical performance parameters or self-assessed parameters was seen beyond 6 months after OLT. In conclusion, these findings indicate that OLT combined with a supervised post-OLT exercise program improves physical fitness, muscle strength, and functional performance in individuals with chronic liver disease. PMID- 10388504 TI - Reconstitution of the actin-scavenger system after orthotopic liver transplantation for end-stage liver disease: a prospective and longitudinal study. AB - Serum levels of the actin scavenger Gc-globulin (group-specific component, vitamin D-binding protein), a member of the albumin multigene family, are decreased in severe liver disease but have not been evaluated in relation to liver transplantation. We measured Gc-globulin and Gc-globulin-actin complex ratio daily for 2 weeks after transplantation in 17 patients with end-stage liver disease. Before transplantation, Gc-globulin levels were significantly less in the patients than in healthy controls (235 +/- 106 v 340 +/- 35 mg/L, respectively; P<.001), whereas complex ratio level was in the normal range. Five patients (group N) had pretransplantation Gc-globulin values within the normal range (mean +/- 2 SD), and 12 patients had subnormal values (group S). In group N, mean Gc-globulin levels posttransplantation remained stable at a lower level than before transplantation but still within normal range. In this group, cold ischemia time correlated inversely with Gc-globulin levels on day 2 (r = -0.88; P <.05). In group S, normal mean levels were reached at a mean of 11 days after transplantation. However, almost half these patients had subnormal Gc-globulin levels at day 14. Complex ratio levels remained normal in the study period in both groups. Prothrombin index levels (plasma coagulation factors II, VII, and X) were identical in both groups and returned to normal 7 days posttransplantation, whereas plasma albumin levels were less than normal in both groups and further decreased after transplantation. In conclusion, the maintenance (group N) or reestablishment (group S) of serum Gc-globulin to normal levels occurred in the early posttransplantation course in the same time frame as the prothrombin index. Gc-globulin synthesis seems unrelated to albumin synthesis. A prolonged cold ischemia time may cause reduced Gc-globulin levels early after transplantation. PMID- 10388505 TI - Health-related quality of life after liver transplantation: a meta-analysis. AB - The goal of this study is to assess health-related quality of life (HRQL) after orthotopic liver transplantation (OLT). Structured MEDLINE and Embase literature searches identified 5473 potentially relevant articles. Thirty-two additional references were collected from the bibliographies. Of the 5505 identified articles, 49 studies reporting data on 3576 transplant recipients met our inclusion criteria, which were an assessment of quality of life (QOL) in adult patients reported as either pretransplantation and posttransplantation data or with a comparison group and written in English. We combined posttransplantation QOL scores from 15 studies that reported data from the same QOL scales to assess the magnitude of the effect of OLT on QOL scales. We also performed a sign test on the 49 studies to evaluate the direction (positive or negative) of the effect of transplantation on QOL. Transplantation resulted in an improvement of 32% in Karnofsky scores, 11% in Sickness Impact Profile scores, and 20% to 50% in the domains of the Nottingham Health Profile. The sign test showed significant improvement in posttransplantation physical health (P <.0004), sexual functioning (P <.008), daily activities (P <.02), general HRQL (P <.02), and social functioning (P <.05), but not psychological health (P <.08). In general, the HRQL of the 3576 patients was impaired pretransplantation and improved posttransplantation. Transplant recipients reported large gains in those aspects of QOL most affected by physical health and smaller improvements in areas affected by psychological functioning. PMID- 10388506 TI - Rapid progression to high-grade dysplasia in Barrett's esophagus after liver transplantation. AB - There is an increased incidence of malignancies in transplant recipients. Accelerated progression from a premalignant lesion to carcinoma has been reported in transplant recipients with skin cancer and colon cancer. Whereas Barrett's esophagus is a common premalignant condition in the normal population, rapid progression to severe dysplasia or carcinoma has not been widely reported in transplant recipients. We report on a liver transplant recipient who developed rapid progression from Barrett's esophagus without dysplasia to high-grade dysplasia within 9 months after transplantation. PMID- 10388507 TI - Pretransplant ursodeoxycholic acid therapy and liver transplantation in patients with primary biliary cirrhosis: win, win, win? PMID- 10388508 TI - Concepts and controversies in perioperative care for liver transplantation: proceedings of a conference sponsored by the International Liver Transplantation society, Orlando, FL, October 1998. PMID- 10388509 TI - Transesophageal echocardiography and orthotopic liver transplantation: general concepts. PMID- 10388511 TI - Anesthesia for liver transplantation: is this generalist or specialist care? PMID- 10388510 TI - Complications of liver transplantation: a perioperative perspective. PMID- 10388512 TI - Basic principles of cerebral protection in humans. PMID- 10388513 TI - Cerebral protection and resuscitation in the liver transplant recipient. PMID- 10388514 TI - Introduction AB - Anogenital warts have become one of the most common sexually transmitted diseases reported in the Western World. The frustration of treatment for both patient and carer is well recognised. Current available methods rely principally on ablation of visible lesions, with hospital-based treatments often requiring multiple attendance by out-patients. Overall, the current failure rate, recurrence rate, and side-effects of these treatments are highly unsatisfactory. Imiquimod, recently launched in the US under the brand name Aldaratrade mark cream, represents the most interesting and innovative approach to therapy to become available in many years. Imiquimod is an immune response modifier, therefore this symposium report addresses the vital issues of the immune based response to human papilloma virus (HPV) infection, as well as the problems of persistence caused by HPV disease. The mechanisms by which imiquimod can induce an inflammatory and cell-mediated response are discussed. Also reviewed are the consequent imiquimod clinical results and the reasons why they show great cause for optimism in HPV treatment. As a home-based effective treatment with a low recurrence rate, imiquimod has already generated enthusiasm on an international scale. This symposium report presents the thoughts and experiences of medical specialists from various countries regarding the treatment of genital HPV infections and the place for imiquimod in clinical practice. PMID- 10388515 TI - Closing remarks AB - Drawing the meeting to a close, co-chairman Dr. Selim Aractingi said that it is now clear that local immune factors are important in determining the regression of warts, and that imiquimod appears to mimic the mechanism of immunity occuring in people in whom warts regress without treatment. He summarised the main presentation points, stating that topical imiquimod 5% - when applied three times each week overnight at home - is an effective therapy with acceptable side effects. Dr. Aractingi noted that the recurrence rate is lower than those reported with most other available wart therapies. Further studies to specifically compare recurrence rates are warranted, he concluded. PMID- 10388516 TI - The HNF-3alpha transcription factor is a primary target for retinoic acid action. AB - We have previously demonstrated that gene expression of the hepatocyte nuclear factor 3alpha (HNF-3alpha) transcription factor is activated during retinoic-acid induced differentiation of F9 embryonal carcinoma cells (A. Jacob et al. (1994). Nucleic Acids Res. 22, 2126-2133). We have extended these studies and now show that HNF-3alpha mRNA is induced approximately 6 h after addition of retinoic acid to the cells, peaks at 1 day postdifferentiation, and then declines to undetectable levels. Furthermore, HNF-3alpha induction occurs in the absence of de novo protein synthesis, suggesting that it is a primary target for retinoic acid action. In order to corroborate this hypothesis, we have mapped the cis acting HNF-3alpha promoter site that mediates the retinoic acid response. DNA sequence analysis indicates that the HNF-3alpha promoter contains an authentic retinoic acid response element (RARE) of the DR5 class. As expected, this element is able to confer retinoic acid responsiveness to a heterologous promoter. In addition, the HNF-3alpha-specific RARE is able to interact with various retinoic acid receptor heterodimers of the RAR/RXR type. Since HNF-3alpha is induced early during mammalian neurogenesis, our data shed new light on the connection between retinoic-acid-mediated HNF-3alpha activation and establishment of the neuronal phenotype. PMID- 10388517 TI - FGF receptor availability regulates skeletal myogenesis. AB - Fibroblast growth factors (FGFs) and their receptors are critical participants in embryonic development, including the genesis of skeletal, cardiac, and smooth muscle. FGF signaling is mediated through interactions between multiple FGF ligands and transmembrane tyrosine kinase receptors, resulting in activation of a number of signal transduction pathways. Skeletal myocytes express FGF ligands and receptors in a coordinated fashion, suggesting that these molecules participate in autocrine signaling in the myocyte. Endogenously produced FGF has been shown to inhibit myogenesis, but the role of FGF receptor availability in directing myocyte proliferation and differentiation has not been established. To determine the contribution of receptor availability to the regulation of myogenesis, receptor availability was either increased by expressing a full-length FGF receptor-1 or decreased by expressing a truncated FGF receptor-1 in cultured skeletal myocytes. Constitutive expression of a full-length FGF receptor-1 increased myocyte proliferation and delayed differentiation. Conversely, a reduction in functional FGF receptor signaling by expression of a truncated FGF receptor-1 decreased proliferation and enhanced differentiation of myocytes. These data demonstrate that FGF receptor availability plays a critical regulatory role in skeletal myogenesis. PMID- 10388518 TI - Conjugated linoleic acid inhibits proliferation and induces apoptosis of normal rat mammary epithelial cells in primary culture. AB - The trace fatty acid conjugated linoleic acid (CLA) inhibits rat mammary carcinogenesis when fed prior to carcinogen during pubertal mammary gland development or during the promotion phase of carcinogenesis. The following studies were done to investigate possible mechanisms of these effects. Using a physiological model for growth and differentiation of normal rat mammary epithelial cell organoids (MEO) in primary culture, we found that CLA, but not linoleic acid (LA), inhibited growth of MEO and that this growth inhibition was mediated both by a reduction in DNA synthesis and a stimulation of apoptosis. The effects of CLA did not appear to be mediated by changes in epithelial protein kinase C (PKC) since neither total activity nor expression nor localization of PKC isoenzymes alpha, beta II, delta, epsilon, eta, or zeta were altered in the epithelium of CLA-fed rats. In contrast, PKCs delta, epsilon, and eta were specifically upregulated and associated with a lipid-like, but acetone-insoluble, fibrillar material found exclusively in adipocytes from CLA-fed rats. Taken together, these observations demonstrate that CLA can act directly to inhibit growth and induce apoptosis of normal MEO and may thus prevent breast cancer by its ability to reduce mammary epithelial density and to inhibit the outgrowth of initiated MEO. Moreover, the changes in mammary adipocyte PKC expression and lipid composition suggest that the adipose stroma may play an important in vivo role in mediating the ability of CLA to inhibit mammary carcinogenesis. PMID- 10388519 TI - Autocrine stimulation of human mammary carcinoma cell proliferation by human growth hormone. AB - Here we have investigated the role of autocrine production of human growth hormone (hGH) in the proliferation of mammary carcinoma cells (MCF-7) in vitro. MCF-7 cells were stably transfected with an expression plasmid encoding the hGH gene, and these cells (designated MCF-hGH) synthesized hGH in the cell and secreted hGH to the medium. For control purposes, a MCF cell line was generated (MCF-MUT) in which the start codon of the hGH gene was disabled, and these cells transcribed the hGH gene without translation to hGH protein. The MCF-hGH cell number increased at a rate significantly greater than that of MCF-MUT under serum free conditions. Autocrine hGH also synergized with 10% serum and insulin-like growth factor-1 but not 17-beta-estradiol to increase cell number. The increased proliferation of MCF-hGH cells in both serum-free and serum-containing media could be completely abrogated by the use of the nonreceptor dimerizing hGH antagonist, hGH-G120R. Increased mitogenesis as a consequence of autocrine production of hGH was prevented by inhibition of either the p38 MAPK or p42/44 MAPK pathways. MCF-hGH cells also possessed a higher level of STAT5 (but not STATs 1 and 3) mediated transcriptional activation in both serum-free and serum containing conditions than MCF-MUT cells. Thus we conclude that hGH can act in an autocrine/paracrine manner in human mammary carcinoma cells to promote cell proliferation and transcriptional activation. PMID- 10388520 TI - Identification of a 55-kDa ezrin-related protein that induces cytoskeletal changes and localizes to the nucleolus. AB - Normal and transformed human cells when stained for ezrin, an F-actin-binding ERM (ezrin/radixin/moesin) family protein, revealed a faint and intense immunofluorescence, respectively. Surprisingly, nuclear staining that was assigned to the nucleolus by confocal laser and immunoelectron microscopy was detected in both cell types and was more prominent in normal cells due to the absence of glistering cytoplasmic fluorescence. By Western analysis the nuclear fraction was seen to have a 55-kDa ezrin-reactive protein that did not react to the antibodies raised against the C-terminus of the protein, suggesting that it may correspond to an endogenously cleaved N-terminus of the protein. Transfections of cells with a cDNA encoding full-length ezrin tagged with green fluorescent protein (GFP) at its N-terminus indeed resulted in two GFP-tagged products corresponding to full-length and 55-kDa endogenously cleaved forms. Transfection with a cDNA encoding approximately 55 kDa of the ezrin N-terminus (N ezrin) showed that it can translocate to the nucleus. N-ezrin transfected cells exhibited irregular cell edges and collapse of actin fibers. Similar changes were seen following microinjection of anti-p81/ezrin antibody, suggesting that N-ezrin may function as a dominant negative competitor of ezrin. These data demonstrate the existence of an N-terminal cleavage form of ezrin that localizes to the nucleolus and that its overexpression induces cytoskeletal changes. PMID- 10388521 TI - Colocalization within the nucleolus of two highly related IFN-induced human nuclear phosphoproteins with nucleolin. AB - We have previously reported the identification of two interferon (IFN)-induced cDNAs which code for two proteins, named 41 and 75, which have homology to a number of proteins involved in regulating gene expression. Here we establish that these cDNAs correspond to in vivo synthesized mRNAs. Expression of the 41 and 75 cDNAs, both in vitro and in vivo, generated proteins of 30 and 68 kDa, respectively. In a variety of mammalian cells, 41 and 75 were found to be located in the nucleus, with 41 being localized to the nucleolus, whereas 75, although it is mainly concentrated at the periphery of the nucleolus, is also found throughout the nucleoplasm. Treatment with interferon results in a translocation of 41 to the periphery of the nucleolus and it is in this region that the two proteins colocalize. 41 and 75 were found to colocalize with nucleolin but not with B23 or fibrillarin, three nucleolar proteins involved in ribosome synthesis. This colocalization was not affected by low concentrations of actinomycin D. In view of this and since 41 and 75 have homology to proteins involved in regulating gene expression, we suggest that, in association with nucleolin, they play a role in ribosome biogenesis. PMID- 10388522 TI - MPP+ inhibits proliferation of PC12 cells by a p21(WAF1/Cip1)-dependent pathway and induces cell death in cells lacking p21(WAF1/Cip1). AB - The molecular and biochemical mode of cell death of dopaminergic neurons in Parkinson's disease (PD) is uncertain. In an attempt at further clarification we studied the effects of 1-methyl-4-phenylpyridinium (MPP+), the active metabolite of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), on dopaminergic PC12 cells. In humans and nonhuman primates MPTP/MPP+ causes a syndrome closely resembling PD. MPP+ toxicity is thought to be mediated by the block of complex I of the mitochondrial electron transport chain. Treatment of undifferentiated PC12 cells with MPP+ primarily inhibited proliferation of PC12 cells and secondarily led to cell death after the depletion of all energy substrates by glycolysis. This cell death showed no morphological characteristics of apoptosis and was not blocked by treatment with caspase inhibitors. The inhibition of cell growth was not dependent on an inhibition of complex I activity since MPP+ also inhibited cell proliferation in SH-SY5Y cells lacking mitochondrial DNA and complex I activity (p0 cells). As shown by flow cytometric analysis, MPP+ induced a block in the G0/G1 to S phase transition that correlated with increased expression of the cyclin-dependent kinase inhibitor p21(WAF1/Cip1) and growth arrest. Since treatment with 1 microM MPP+ caused apoptotic cell death in p21(WAF1/Cip1) deficient (p21(-/-)) but not in parental (p21(+/+)) mouse embryo fibroblasts, our data suggest that in an early phase MPP+-induced p21(WAF1/Cip1) expression leads to growth arrest and prevents apoptosis until energy depletion finally leads to a nonapoptotic cell death. PMID- 10388523 TI - A src-related kinase in the brush border membranes of gastrointestinal cells is regulated by c-met. AB - Hepatocyte growth factor (HGF) elicits pleiotropic cellular responses by binding to c-met, a PTK transmembrane receptor. The recent identification of HGF in fluids which enter the gut lumen suggests a mechanism by which c-met molecules are accessible to ligand that is present near the apical surfaces of polarized enterocytes. A subset of c-met molecules was detected, by confocal and immunoelectron microscopic analysis, which colocalizes with a recently identified src-related gastrointestinal tyrosine kinase (gtk) in the brush border membranes of enterocytes. Furthermore, treatment of c-met/gtk-transfected cells with a chemical cross-linking agent revealed that c-met forms a physical complex with gtk, in vivo. Not surprisingly, activation of the receptor molecules with HGF rapidly stimulated gtk enzymatic activity. Similarly, the stimulation of gtk activity occurred when nontransfected primary hepatocytes were exposed to ligand. These findings suggest a model in which HGF binding to luminally accessible c-met stimulates gtk activity. This brush border-associated c-met-linked pathway may be associated with a defined set of epithelial cell responses. PMID- 10388524 TI - Differential effects of the insulin-like growth factor I receptor on radiosensitivity and spontaneous necrosis formation of human glioblastoma cells grown in multicellular spheroids. AB - The purpose of this study is to investigate how the insulin-like growth factor I receptor (IGF-IR) affects cellular radiosensitivity when cells are cultured under different growth conditions. For this, A7(R) and A7(puro) cells were established from human glioblastoma GB A7 cells. The former were derived from the parent cells by stable cotransfection with plasmids carrying human IGF-IR cDNA and a puromycin resistance gene and the latter had the marker gene alone. The cells were either grown exponentially in monolayer cultures or grown in multicellular spheroids as an in vitro model for solid tumors. Spheroids were formed in the two different methods, liquid-overlay (LOC) and spinner (SPC) cultures. Although the growth rate of both cell lines in monolayer was exactly the same, the growth rate of A7(R) spheroids formed in LOC was higher than that of A7(puro) spheroids. A central necrosis region was histologically observed in A7(puro) spheroids, but the corresponding region in A7(R) spheroids was almost completely filled with intact cells in both LOC and SPC spheroids. Both cell lines showed the same radiosensitivity in monolayer cultures in terms of cell viability and clonogenic cell survival. When the spheroids formed in LOC were X-irradiated, the radiosensitivity of A7(R) and A7(puro) cells assayed for cellular clonogenicity was also the same. However, in the spheroids formed in SPC, A7(R) cells were significantly more radiosensitive than A7(puro) cells. The results indicate that overexpression of the IGF-IR could induce radiosensitization of human tumor cells in spheroids while inhibiting spontaneous necrosis formation. This may open a possibility to explore the novel function of the IGF-IR. PMID- 10388525 TI - Induction of interleukin-15 production by HIV-1 nef protein: a role in the proliferation of uninfected cells. AB - Several recent reports have provided evidence that Nef enhances human immunodeficiency virus HIV infectivity, and in vitro experiments with the nef gene have demonstrated the possible role of Nef in modulating immune responses. Exogenous Nef has been demonstrated to induce proliferation of normal human peripheral blood mononuclear cells (PBMC) and to enhance HIV-1 replication. The aim of this study was to evaluate the biological mechanisms by which Nef, used as exogenous protein, modulates cellular activation. We showed that exogenous Nef protein induces the proliferation of unstimulated and suboptimally stimulated normal human PBMC, while it has no effect on the proliferation of optimally stimulated PBMC. Moreover, the activating effect of exogenous Nef on PBMC proliferation was associated with an increase of IFN-gamma, TNF-alpha, and IL-6 production, while, surprisingly, IL-2 production was not affected by Nef. More importantly we showed, for the first time, that Nef exerts its activating effects on PBMC proliferation through IL-15 synthesis induction by monocyte/macrophage population. In conclusion, we found that exogenous Nef protein (i) induces activation of normal PBMC, increasing their proliferative response; (ii) modulates cytokine production; (iii) exerts its activating effects through IL-15 synthesis induction; and (iv) exerts these effects entering monocyte/macrophages. Our results might suggest that Nef enhances the rate of viral replication by a novel mechanism involving the production of IL-15. PMID- 10388526 TI - Matrix metalloproteinase-19 in capillary endothelial cells: expression in acutely, but not in chronically, inflamed synovium. AB - Matrix metalloproteinase-19 (MMP-19), originally isolated as an autoantigen from the synovium of a patient suffering from rheumatoid arthritis (RA), is expressed in smooth muscle cells of the tunica media of large blood vessels of an RA patient, but not in the endothelial cell layer. By contrast, in acutely inflamed tissue, synovial capillaries strongly express MMP-19 in the cytoplasm, as shown by immunofluorescence of cryostat sections. In MMP-19-producing capillaries the beta3 integrin chain was found at the endothelial cell surface, as was the vascular endothelial cell growth factor receptor-2 (KDR). The specific tissue inhibitor of metalloproteinases TIMP-1 was absent or faintly stained in MMP-19 expressing capillaries, whereas TIMP-1, but not TIMP-2, was strongly expressed in large vessels and in MMP-19-negative capillaries of RA synovia. In the spontaneously transformed human umbilical vein endothelial cell line ECV304 neither MMP-19 transcripts nor protein could be detected. By contrast, primary cultures of human endothelial cells of either dermal or adipose tissue origin produced MMP-19 mRNA and protein. The results strongly suggest the regulated induction of matrix metalloproteinase-19 in capillary endothelial cells during acute inflammation and hint at a role of MMP-19 in angiogenesis. PMID- 10388527 TI - Differential responses of proliferating versus quiescent cells to adriamycin. AB - The relative sensitivity of proliferating and quiescent cells to DNA-damaging agents is a key factor for cancer chemotherapy. Here we undertook a reevaluation of the way that proliferating and quiescent cells differ in their responses and fate to adriamycin-induced damage. Distinct types of assays that measure membrane integrity, metabolic activity, cell size, DNA content, and the ability to proliferate were used to compare growing and quiescent Swiss3T3 fibroblasts after adriamycin treatment. We found that immediately after adriamycin treatment of growing cells, p53 and p21(Cip1/Waf1) were induced but the cells remained viable. In contrast, less p53 and p21(Cip1/Waf1) were induced in quiescent cells after adriamycin treatment, but the cells were more prone to immediate cell death, possibly involving apoptosis. Adriamycin induced a G2/M cell cycle arrest in growing cells and a concomitant increase in cell size. In contrast, adriamycin induced an increase in sub-G1 DNA content in quiescent cells and a decrease in cell size. In contrast to the short-term responses, adriamycin-treated quiescent cells have a better long-term survival and proliferation potential than adriamycin-treated growing cells in colony formation assays. These data suggest that proliferating and resting cells are remarkably different in their short-term and long-term responses to adriamycin. PMID- 10388528 TI - Characterization of NF-L and betaIISigma1-spectrin interaction in live cells. AB - Neurofilaments (NFs) are neuron-specific intermediate filaments (IFs) composed of three different subunits, NF-L, NF-M, and NF-H. NFs move down the axon with the slow component of axonal transport, together with microtubules, microfilaments, and alphaII/betaII-spectrin (nonerythroid spectrin or fodrin). It has been shown that alphaII/betaII-spectrin is closely associated with NFs in vivo and that betaII-spectrin subunit binds to NF-L filaments in vitro. In the present study we seek to elucidate the relationship between NF-L and betaII-spectrin in vivo. We transiently transfected full-length NF-L and carboxyl-terminal deleted NF-L mutants in SW13 Cl.2 Vim- cells, which lack an endogenous IF network and express alphaII/betaIISigma1-spectrin. Double-immunofluorescence and electron microscopy studies showed that a large portion of betaIISigma1-spectrin colocalizes with the structures formed by NF-L proteins. We found a similar association between NF-L proteins and actin. However, coimmunoprecipitation experiments in transfected cells and the yeast two-hybrid system results failed to demonstrate a direct interaction of NF-L with betaIISigma1-spectrin in vivo. The presence of another protein that acts as a bridge between the membrane skeleton and neurofilaments or modulating their association may therefore be required. PMID- 10388529 TI - A role for gelsolin in stress fiber-dependent cell contraction. AB - Gelsolin is an abundant actin binding protein that mediates the rapid remodeling of cortical actin filaments through severing, capping, and nucleating activities. Most of the attention on the intracellular function of gelsolin has focused on the remodeling of the cortical actin meshwork but the localization of gelsolin to other regions of the cell suggests that it may have other important functions elsewhere. In cultured fibroblasts, gelsolin is heavily enriched in stress fibers, where its function in these contractile organelles is unknown. To study gelsolin function during stress fiber formation and cell contraction, we first assessed gelsolin levels in stress fiber preparations from lysophosphatidic acid (LPA)-treated human fibroblasts. LPA induced a large, time-dependent, calcium independent increase of actin, gelsolin, alpha-actinin, and tropomyosin in stress fiber preparations. A microinjected gelsolin antibody that inhibits severing by gelsolin reduced stress fibers. Anti-sense-transfected gelsolin-depleted 3T3 cell lines treated with LPA after serum starvation required approximately 6 h to form stress fibers and focal adhesions, in contrast to control lines transfected with vector only, which formed stress fibers 15 min after addition of LPA. In cells microinjected with the gelsolin antibody that inhibits severing, Mg-ATP-induced cell contraction was greatly reduced in approximately 90% of injected cells compared to cells injected with an irrelevant antibody. Gelsolin-depleted cells were incapable of collagen gel contraction and showed no apparent Mg-ATP-induced cell contraction compared to cell lines transfected with vector only. The involvement of gelsolin in cell contraction and remodeling of collagen gels suggests a novel role for gelsolin in stress fiber-dependent cell function. PMID- 10388530 TI - Gp38k, a protein synthesized by vascular smooth muscle cells, stimulates directional migration of human umbilical vein endothelial cells. AB - Gp38k is a 383-amino-acid secreted glycoprotein expressed by cultured vascular smooth muscle cells during the time of transition from a proliferating monolayer culture to a nonproliferating multilayered (differentiated) culture. Expression continues as the cell culture forms multicellular nodules. Because this transition period involves active cell migration, we evaluated the effects of exogenously added gp38k on vascular endothelial cell (HUVEC) migration and chemotaxis. Here we demonstrate that gp38k acts as a chemoattractant for HUVECs and stimulates cell migration in Boyden chambers at a level comparable to that achieved with the known endothelial cell chemoattractant bFGF. The migration effect is neutralized by the presence of a polyclonal anti-gp38k antibody. Because gp38k expression is also correlated with changes in culture morphology, we also assessed its ability to act as an agonist of HUVEC morphology using cultures growing on Matrigel. We report that gp38k stimulates endothelial cell tubulogenesis in this assay system. These results provide the first evidence that gp38k may function in angiogenesis by stimulating the migration and reorganization of vascular endothelial cells. PMID- 10388531 TI - Phenotype dictates the growth response of vascular smooth muscle cells to pulse pressure in vitro. AB - The objective of this study was to determine the effect of phenotype on pulse pressure-induced signaling and growth of vascular smooth muscle cells in vitro. Using a perfused transcapillary culture system, cells were exposed to increases in pulsatile flow and hence pulse pressure and maintained for 72 h before cells were harvested. Cell proliferation was determined by cell number, DNA synthesis, and proliferating cell nuclear antigen expression. Mitogen-activated protein kinase (MAPK) levels were determined by immunoblot and kinase activity by phosphorylation of myelin basic protein. Cell phenotype was determined by immunoblot and immunocytofluorescence using antisera specific for the differentiation markers alpha-actin, myosin, calponin, osteopontin, and phospholamban. In cells that highly expressed these differentiation markers, there was a significant increase in cell growth in response to chronic increases in pulse pressure without a significant change in MAPK activity in these cells. In contrast, in cells that weakly expressed SMC differentiation markers, there was a significant decrease in cell growth concomitant with a significant decrease in MAPK signaling in these cells. We conclude that SMC phenotype dictates the growth response of SMC to mechanical force in vitro. PMID- 10388532 TI - Role of epidermal growth factor receptor in basal and stimulated colonic epithelial cell migration in vitro. AB - Colonic mucosal wounds are repaired, in part, by epithelial migration. Signaling mechanisms regulating this migration are poorly characterized. This study aimed to examine the role that the epidermal growth factor (EGF) receptor (EGF-R) and its ligands, EGF and transforming growth factor-alpha (TGF-alpha), play in migration in wounded in vitro models of colonic epithelium. Migration was assessed over 24 h in circular wounds made in confluent monolayers of LIM1215 human colon cancer cells. EGF and TGF-alpha stimulated migration twofold from 4 h after wounding. Basal migration and the motogenic effects of short chain fatty acids and hepatocyte growth factor were mediated through enhanced binding of TGF alpha to EGF-R, while trefoil peptide-mediated motogenesis required EGF-R activation independently of TGF-alpha binding. Activation of protein kinase C (PKC) stimulated migration, an effect more potent than, and independent of, EGF-R activation. However, neither inhibition of PKC by Ro 31-8220 nor depletion of PKC by pretreatement with phorbol myristate acetate attenuated EGF-R-mediated motogenesis. In conclusion, EGF-R activation via TGF-alpha binding, or intracellularly, mediates basal LIM1215 migration and the effects of several motogens, with the exception of PKC activators. Since EGF-R and PKC have physiological activators in vivo, they may control colonic mucosal repair processes following injury. PMID- 10388533 TI - Apoptosis of Caco-2 intestinal cells invaded by Listeria monocytogenes: protective effect of lactoferrin. AB - The apoptosis of infected hepatocytes is a critical step in nonspecific defense against Listeria monocytogenes infection. We have observed that infection by L. monocytogenes in enterocyte-like cells (Caco-2) results in apoptosis. However, a large fraction of infected intestinal epithelial cells escape from cellular condensation and fragmentation, typical of programmed cell death, and become necrotic. The balance between apoptosis and necrosis seems to be influenced by the number of internalized bacteria. The presence of 1 mg/ml of bovine lactoferrin, an iron-binding protein, added to monolayers before the bacterial infection, decreases the number of internalized bacteria and therefore the overall number of dead cells, and, more importantly, all dead cells are killed by apoptosis and not necrosis. PMID- 10388534 TI - Caspase-8 mediates caspase-3 activation and cytochrome c release during singlet oxygen-induced apoptosis of HL-60 cells. AB - We reported previously that singlet oxygen, generated by irradiation of rose bengal with visible light, induced apoptosis in human promyelocytic leukemia HL 60 cells. However, the mechanism of apoptosis caused by this reactive oxygen species is unclear. In this study, we demonstrate that singlet oxygen induced caspase-3 activation and Z-DEVD-FMK, a caspase-3 inhibitor, blocked apoptosis induction, while caspase-1 activity was not detectable and the caspase-1 inhibitor Z-YVAD-FMK had a very limited effect on apoptosis. This suggests that the activation of caspase-3 by singlet oxygen is essential for the commitment of cells to undergo apoptosis. Further studies showed that singlet oxygen induced an increase in caspase-8 activity and a reduction in mitochondrial cytochrome c. Time course analysis indicated that the cleavage of caspase-8 precedes that of caspase-3. In addition, blockade of caspase-8 by Z-IETD-FMK inhibited cleavage of pro-caspase-3 and prevented loss of mitochondrial cytochrome c. These results suggest that caspase-8 mediates caspase-3 activation and cytochrome c release during singlet oxygen-induced apoptosis in HL-60 cells. PMID- 10388535 TI - TGF-beta1 inhibits NF-kappaB activity through induction of IkappaB-alpha expression in human salivary gland cells: a possible mechanism of growth suppression by TGF-beta1. AB - Transforming growth factor (TGF)-beta is the prototype of a large superfamily of signaling molecules involved in the inhibition of proliferation of multiple epithelial cell types. Although accumulated evidence indicates the mechanisms of the antimitogenic effect of TGF-beta in a variety of cell types, the signal transduction mechanism underlying the regulation of NF-kappaB transcription factor by TGF-beta is largely unknown. Because NF-kappaB is not only involved in inflammatory responses but also mediates cell growth, we have investigated the effect of TGF-beta1 on the activity of NF-kappaB and the role of the inhibitory IkappaB-alpha protein in the growth of the human salivary gland cell clones NS-SV AC, HSGc, and cl-1. NF-kappaB, which is usually maintained in an inactive state by protein-protein interaction with IkappaB, was found to be constitutively active in salivary gland cell lines. Upon treatment of cell clones with TGF beta1, the NF-kappaB activity in NS-SV-AC and HSGc, but not in cl-1, which lacks the expression of TGF-beta type II receptor, was suppressed. In NS-SV-AC and HSGc, this inhibition was mediated by the induction of IkappaB-alpha at the mRNA and protein levels. The blocking of NF-kappaB subunit with a specific antisense oligonucleotide reduced the growth rate of all of the cell clones, including cl 1. Introduction of a mutated form of IkappaB-alpha cDNA into NS-SV-AC suppressed the growth rate of this cell clone. These results indicate that TGF-beta1 downregulates NF-kappaB activity through the induction of IkappaB-alpha expression in human salivary gland cells and that inhibition of NF-kappaB activity suppresses the growth rate of these cells. PMID- 10388536 TI - Increased phosphorylation of eukaryotic initiation factor 2alpha at the G2/M boundary in human osteosarcoma cells correlates with deglycosylation of p67 and a decreased rate of protein synthesis. AB - The rate of protein synthesis in higher eukaryotes is largely regulated at the level of eIF2alpha phosphorylation by its kinases. A cellular glycoprotein, p67, protects eIF2alpha from phosphorylation. An enzyme, p67-deglycosylase, when active, removes the carbohydrate moieties from p67 and inactivates it. Subsequently, protein synthesis is inhibited. During mitosis the overall rate of protein synthesis sharply declines. To understand the molecular mechanism underlying this inhibition of protein synthesis, we have examined the phosphorylation of eIF2alpha and the activity of p67. We find that the phosphorylation of eIF2alpha increases at the G2/M border of cycling U2-OS cells, and p67 is deglycosylated at the same period of the cell cycle. In addition, the level and the activity of p67-deglycosylase also increase at the G2/M boundary of cycling U2-OS cells. These results thus provide an important in vivo correlation between the increased phosphorylation of eIF2alpha and deglycosylation of p67 by p67-deglycosylase at the G2/M boundary of cycling U2-OS cells. This may explain in part the inhibition of protein synthesis in U2-OS cells approaching mitosis. PMID- 10388537 TI - Urokinase-type plasminogen activator binding to its receptor stimulates tumor cell migration by enhancing integrin-mediated signal transduction. AB - Urokinase-type plasminogen activator (uPA) and its receptor (uPAR) participate in matrix degradation and cell migration by focusing proteolysis and functioning as a signaling ligand/receptor complex. uPAR, anchored by a lipid moiety in the membrane, is thought to require a transmembrane adapter to transduce signals into the cytoplasm. To study uPAR signaling, we transfected the prostate carcinoma cell line LNCaP, which does not express endogenous uPA or uPAR, with a uPAR encoding cDNA, resulting in high-level surface expression. We studied migration of these cells on fibronectin, which is mediated by the integrin alpha5beta1. Ligation of uPAR with uPA or its amino-terminal fragment enhanced haptotactic migration to fibronectin. In cells on fibronectin, but not on poly-l-lysine, ligation of uPAR also resulted in tyrosine phosphorylation of several proteins, including two proteins involved in integrin signaling, focal adhesion kinase and the crk-associated substrate p130(Cas). Furthermore, after uPAR ligation, uPAR was co-immunoprecipitated with beta1 integrins from the detergent-insoluble fraction of cell lysates. Thus, our data suggest that uPAR occupancy results in an interaction between uPAR and integrins and a potentiation of integrin-mediated signaling, which leads to enhanced cell migration. PMID- 10388538 TI - Interferon-gamma inhibits CD44-hyaluronan interactions in normal human B lymphocytes. AB - The interaction of CD44 with its ligand hyaluronan (HA) plays a vital role in lymphopoiesis, lymphocyte homing, T cell activation, and metastasis. This study addresses the effect of cytokines involved in B cell growth on CD44-HA interactions in normal human B cells. Activation of B lymphocytes with LPS, pokeweed mitogen, or anti-IgM antibodies with or without IL-2 or IL-4 failed to induce HA adhesion. Stimulation of B cells with the phorbol ester PMA, however, induced strong HA recognition, which was inhibited by IFN-gamma and to some extent by IL-4. Investigation of the potential molecular mechanism involved revealed that PMA-induced HA adhesion correlated with enhanced expression of CD44 H- and V6-containing isoforms, as determined by flow cytometry, and the differential induction of V4- and V5-containing isoforms, as determined by reverse transcriptase-based polymerase chain reaction analysis. The inhibition of PMA-induced adhesion by IFN-gamma and IL-4 correlated with the downregulation of CD44 H expression and altered usage of exons V4 and V5. However, changes in the electrophoretic mobility of CD44 proteins, as a measure of posttranslational modifications, were not detected in response to PMA and IFN-gamma or PMA and IL 4. These results suggest that the inhibition of PMA-induced HA adhesion by IFN gamma and IL-4 may influence B cell migration through their ability to downregulate CD44-HA interactions. PMID- 10388539 TI - Antisense inhibition of protein kinase Calpha reverses the transformed phenotype in human lung carcinoma cells. AB - The protein kinase C (PKC) family, which functions through serine/threonine kinase activity, is involved in signal transduction pathways necessary for cell proliferation and differentiation. Its critical role in processes relevant to neoplastic transformation and tumor invasion renders PKC a potentially suitable target for anticancer therapy. To explore whether antisense blocking of PKCalpha would inhibit the neoplastic properties in tumor cells, human lung carcinoma LTEPa-2 cells were transfected with a recombinant plasmid, pXJ41-CKPalpha, with PKCalpha cDNA inserted in the antisense orientation. In LT.AS4 cell clones stably expressing antisense PKCalpha mRNA, the amounts of PKCalpha protein and total PKC activity were decreased when compared to control cells. The expression of antisense PKCalpha markedly inhibited the cell proliferation rate, colony forming efficiency in soft agar, and tumorigenecity in nude mice. Furthermore, the mRNA levels of oncogenes (Ha-ras, c-jun, and c-fos) were seen to decrease to varying degrees. Reduced DNA binding activity of transcription factor AP-1 was also observed using gel shift analysis, suggesting that one major molecular mechanism by which PKCalpha can exert its effects on cell growth and transformation is through regulation of AP-1 transcription factor activity. Taken together, these data provide evidence for the ability of antisense PKCalpha expression to reverse the transformed phenotype of human lung carcinoma cells and support the development of PKCalpha inhibitors for the clinical treatment of cancers. PMID- 10388542 TI - Ultrastructural and cytochemical characterization of aphid invasion by the hyphomycete verticillium lecanii AB - Chronological events of the interaction between the hyphomycete Verticillium lecanii and the potato aphid Macrosiphum euphorbiae were investigated by light, scanning, and transmission electron microscopy. The parasitism of M. euphorbiae by V. lecanii appears to involve the following events: (i) adherence of conidia to the host cuticle through a thin mucilagenous matrix; (ii) germination of the conidia and production of mycelium that colonizes the surface of the cuticle; (iii) penetration of germ tubes into the aphid cuticle 24 h after application of the pathogen; (iv) extensive lateral development of hyphae accompanied by pronounced degradation of the cuticular layers by 72 h. Labeling with the WGA/ovomucoid-gold complex showed that penetration and colonization of the cuticle by the fungus resulted from localized enzymatic hydrolysis, likely through the synergistic action of chitinases and mechanical pressure; (v) production of blastospores and massive invasion of aphid internal tissues; (vi) assimilation of nutrients and accumulation of lipids by fungal cells; and (vii) production of conidiophores and release of the fungus from aphid cadavers. These observations bring further insights into the mechanisms by which V. lecanii parasitizes M. euphorbiae. They also provide a basis for comparing the modes of action of V. lecanii against hosts from fungal and animal origins. Copyright 1999 Academic Press. PMID- 10388540 TI - The role of scavenger receptor class A in the adhesion of cells is dependent on cell type and cellular activation state. AB - Scavenger receptor class A (SR-A) facilitates the development of atherosclerosis, which might be due to its role in the uptake of modified low-density lipoproteins. However, the receptor is also suggested to be important for cell adhesion, thereby potentially influencing the residence time of cells in vivo. Using SR-A-deficient mice, we investigated the role of SR-A in the adhesion of peritoneal macrophages (PM) and tissue macrophages (Kupffer cells). In resident PM no effect of the absence or presence of SR-A on cell adhesion was observed, either in the presence or in the absence of serum. However, in thioglycollate induced PM, SR-A is important for adhesion both in the presence and in the absence of serum and more than 85% of the divalent-cation-independent adhesion in the presence of serum is mediated by SR-A. In unactivated Kupffer cells, like in resident PM, adhesion is not influenced by the absence or presence of SR-A. In vivo administration of phorbol 12-myristate 13-acetate leads to the activation of Kupffer cells, and it appears that under these conditions SR-A does contribute to adhesion, since both in the absence and in the presence of serum SR-A is responsible for about 35% of cell adhesion. It is concluded that SR-A is important for the divalent-cation-independent adhesion of activated PM and Kupffer cells, suggesting that SR-A may influence the residence time of cells at sites of cellular activation, e.g., in atherosclerotic plaques and during liver infection. PMID- 10388543 TI - Histopathological changes after induced injury in leeches. AB - Cells involved in leech inflammatory responses have been characterized by morphological, histochemical, and immunohistochemical methods. Macrophage-like cells, NK-like cells, and granulocytes migrated shortly after injury by pricking with bacterial lipopolysaccharide. Inflammatory responses increased progressively and provoked cell migration to the body wall and then to wound surfaces. Macrophages, NK cells, and granulocytes display similar features and behavior traits in invertebrates and vertebrates. PMID- 10388544 TI - Nosema tyriae n.sp. and Nosema sp., microsporidian parasites of Cinnabar moth Tyria jacobaeae. AB - Nosema tyriae n.sp. was found in 63% of a population of Cinnabar moth larvae (Tyria jacobaeae). The infection was found in the gut wall, silk glands, and fat body and was probably generalized but appeared to be of low pathogenicity. Merogony and sporogony were by binary fission of diplokaryotic stages. Fresh spores were elongate, slightly pointed at the anterior end, and measured 4.7 x 2.0 microm. Ultrastructural features of special interest were 20-nm tubules connecting the surface of sporonts with host cell cytoplasm and, in the spores, a deeply domed polar sac, polaroplast consisting of closely packed longitudinally arranged membranes and loosely packed horizontally arranged membranes, and 10.5 14 coils of the polar tube in a single rank. The 16S rRNA genes of N. tyriae and Nosema bombycis from silkworms, Bombyx mori, differed by only six nucleotides and N. tyriae spores gave a moderately positive reaction with a monoclonal antibody raised to N. bombycis. N. tyriae was infective to B. mori but was less virulent than N. bombycis. However, no amplification product was obtained by PCR using N. tyriae DNA and primers considered to be specific for N. bombycis. Also, the spores of the two species are of entirely different shapes. A second diplokaryotic microsporidium, Nosema sp., found as a light infection in only one of the larvae had much smaller developmental stages and spores measuring 3.8 x 2.0 microm (fixed). Ultrastructurally it was distinguished by an abundance of dense membranes in cytoplasmic vesicles in both meronts and sporonts. Spores with up to 15 coils of the polar tube in irregular clusters or with about 12 coils in a single rank were observed in the tissues fixed from the one larva infected with this parasite. As this larva had been kept with N. tyriae-infected larvae for a few days before examination, it is possible that the two types of spores resulted from a double infection. PMID- 10388545 TI - Impact of bacillus thuringiensis var. israelensis On larvae of chironomus thummi thummi and psectrocladius psilopterus (Diptera: chironomidae) AB - Chironomid larvae, especially species of the subfamily Chironominae, are known to be sensitive to the mosquitocidal bacterium Bacillus thuringiensis var. israelensis (B.t.i.). In this study, bioassays and electron microscopic investigations were carried out with third- and fourth-instar larvae of Chironomus thummi thummi Kieffer (subfamily Chironominae) and Psectrocladius psilopterus Kieffer (subfamily Orthocladiinae) in order to study the sensitivity of species belonging to different chironomid subfamilies. Both species showed susceptibility to increased B.t.i. concentrations, with LC50 values (24 h) ranging from about 40- to 60-fold the LC50 for Aedes aegypti (LC50 (24 h) is 0.77 mg/L for C. thummi thummi and 1.17 mg/L for P. psilopterus). C. thummi thummi was shown to be twice as sensitive as P. psilopterus. Ultrastructural investigations of the anterior midgut showed cellular alterations in larvae exposed to a high B.t.i. concentration (2.8 mg/L, about 50-fold the LC50 for A. aegypti), such as swelling of mitochondria, dilatation of intercellular spaces and basal labyrinth, fenestration or disorganization of the Golgi complex, concentric arrangement of rough endoplasmic reticulum, and an increase of lysosomes and myelin figures. Electron-lucent regions within the cell, cell protrusion, and, in some cases, swelling or lysis of cells were further effects observed in treated animals. Most effects were found in both species, though they seemed to be more severe in C. thummi thummi. The alterations coincide with those known from target organisms (Culicidae, Simuliidae). This study shows that there is a difference in sensitivity to B.t.i. between chironomid species from different subfamilies and that the susceptibility of chironomid larvae to the bacterial toxins is due to damage of the midgut epithelium as it is in target organisms. Copyright 1999 Academic Press. PMID- 10388546 TI - Occurrence and virulence of a granulosis virus in phthorimaea operculella (Lep., gelechiidae) populations in indonesia AB - Indonesian potato growers face increasing problems from Phthorimaea operculella, whose larvae are responsible for damage in potato tubers. Use of biological control agents, and specifically entomopathogenic micro-organisms, could be an alternative method to chemical control. From this perspective, we carried out a screening and an evaluation of local granuloviruses which are naturally present in Indonesian pest populations. The use of an enzyme-linked immuno-sorbent assay allowed us to isolate three granulovirus strains from P. operculella larvae collected from three different locations in Indonesia: Wonsosobo (Central Java), Lembang (West Java), and Berastagi (Northern Sumatra). Bioassays did not reveal significant differences in the biological properties (LC50 and ST50) of these strains. The restriction pattern of the viral genome (Wonsosobo strain) presented only minor variations compared to other isolates collected in different regions of the world. Copyright 1999 Academic Press. PMID- 10388547 TI - Laboratory screening of nematodes isolated from south australia for potential as biocontrol agents of helicid snails AB - The helicid snails Cernuella virgata, Theba pisana, Cochlicella acuta, and Cochlicella barbara are introduced pests in grain crops and pastures in southern Australia. A survey for nematodes with potential for biocontrol of these snails was carried out and six local nematode isolates were tested for their ability to kill C. virgata, T. pisana, and C. acuta using a soil-based laboratory bioassay. The rhabditid isolate R 954 was highly pathogenic to all three snail species with mortality increasing with increasing nematode densities and exposure time. C. acuta was most susceptible to R 954. In addition, high mortalities followed exposure of C. acuta to two cephalobid isolates, C 916 and C 920. Snail mortality caused by the other nematode isolates tested was too low for further consideration as possible biocontrol agents. Copyright 1999 Academic Press. PMID- 10388548 TI - A new picorna-like virus, PnPV, isolated from ficus transparent wing moth, Perina nuda (Fabricius). AB - Two viruses, Perina nuda nucleopolyhedrovirus and a new picorna-like virus, were previously isolated from P. nuda larvae with flacherie. In this study the new picorna-like virus was characterized using physical and biochemical methods. This small virus appears to belong to the family Picornaviridae and we propose the name PnPV. PnPV can be propagated in its homogenous cell line, NTU-PN-HH. PnPV purified from the cell line resembles PnPV isolated from insects: under electron microscopy, it exhibits icosahedral symmetry, measures 30 nm in diameter, and has no envelope and no distinct surface structure in negatively stained preparations. In addition, we show here that PnPV has a buoyant density of 1.381 g/ml in cesium chloride, the viral genome was composed of one single-stranded RNA molecule with a length of 10 kb, and poly(A) tract and polyacrylamide gel electrophoresis of purified viral particles revealed three major (31.5, 29.7, and 28.4 kDa) and three minor (27. 0, 24.5, and 4.0 kDa) structural proteins. PMID- 10388549 TI - Effects of fungal infection on the alarm response of pea aphids AB - Pea aphids (Acyrthosiphon pisum, Homoptera: Aphididae) infected with the fungal pathogen Erynia neoaphidis (Zygomycetes: Entomophthorales) were less sensitive to the aphid alarm pheromone (E)-beta-farnesene than uninfected aphids. Approximately 83% of the healthy aphids and 45% of the infected aphids (2 and 3 days post-inoculation) responded to the alarm pheromone. The percentage of nonresponding aphids increased as the disease progressed. When squeezed (simulated attack) healthy aphids and aphids at an advanced stage of infection elicited a response similar to that of uninfected aphids, suggesting that the alarm pheromone was produced by diseased insects. Aphids infected for 2 or 3 days did not recolonize the upper regions of bean plants after dislodgment. This showed that infected aphids are less active at late stages of infection. The implications of these results are discussed with respect to fungal transmission and biological control. Copyright 1999 Academic Press. PMID- 10388550 TI - Effects of the protozoan parasite ophryocystis elektroscirrha on the fitness of monarch butterflies (Danaus plexippus) AB - We evaluated the effects of the protozoan parasite Ophryocystis elektroscirrha on the survival and reproduction of monarch butterflies. Because larvae in natural populations are likely to experience a wide range of natural parasite population densities, we examined the effects of increasing spore density (0, 10, 100, or 1000 spores per larva) on host fitness. Parasites had little effect on monarch survival or reproduction, except at the highest dose. Monarchs inoculated with 1000 spores per larva had decreased survival to eclosion, and this effect was more severe when larvae were inoculated at an earlier stage (first versus third instar). Monarchs inoculated with higher spore densities also emerged with smaller wingspans and lower body mass than noninoculated adults. Infection with the highest dose of O. elektroscirrha led to decreased male lifespan and reproductive success, but females infected with O. elektroscirrha did not experience a significant decline in lifetime fecundity. However, heavily infected females in outdoor enclosures were less active than uninfected females and gained weight during their adult lifespan. Among samples of adult monarchs captured in natural populations, parasite loads were associated with butterfly condition and activity. Heavily infected adults captured breeding in western North America and southern Florida were smaller than uninfected monarchs. Among overwintering adults in Mexico and California, mating activity was positively associated with higher parasite loads. In addition, the proportion of adults with low and intermediate spore loads (as opposed to no spores) was higher among adults with greater wing tatter and scale loss. Our findings of minor effects of O. elektroscirrha on the survival and reproduction of monarch butterflies are consistent with the expectation that maternally transmitted parasites should have little or no effect on host fitness compared with horizontally transmitted parasites. However, because our laboratory studies demonstrated that monarchs exposed to the highest parasite density experienced decreased larval survival, smaller adult size, and shorter adult lifespans, additional transmission routes are likely to be important for parasite maintenance in natural populations. Copyright 1999 Academic Press. PMID- 10388551 TI - New north american records of aquatic insects as paratenic hosts of pheromermis (Nematoda: mermithidae) AB - Several species of aquatic insects in Trout Park Nature Preserve (Elgin, IL) were observed to have small, black spots (<0.1 mm diameter) visible within their bodies. Microscopic examination revealed these spots to be coiled juveniles of a mermithid (Nematoda: Mermithidae). Based on host habitat (seepage areas and rivulets), host species (aquatic insects), and size (mean diameter of coiled juveniles = 79 &mgr;m), it is likely that these mermithids were in the genus Pheromermis. Since adult mermithids were not found, species determination was not feasible, and the possibility of a new species cannot be ruled out. Pheromermis pachysoma and Pheromermis vesparum, however, are two species known to use aquatic insects as paratenic (i.e., transport) hosts in order to reach their definitive hosts, vespid wasps. Wasp larvae are infected by consuming the flesh of adult aquatic insects that contain the coiled juveniles of these Pheromermis spp. Of the 19 macroinvertebrate species examined in this study, Pheromermis juveniles were found in 4 caddisfly species (Hesperophylax designatus, Lepidostoma liba, Glossosoma intermedium, and Diplectrona modesta) and in 2 stonefly species (Clioperla clio and Amphinemura delosa). In addition to all 6 insect species being new host records for Pheromermis infection, this also represents the first report of nematode infection in stoneflies within the Western Hemisphere and of a Pheromermis sp. in Illinois. Among trophic groups, insect detritivores have been frequently recorded infected with coiled Pheromermis juveniles because of their direct consumption of eggs, and we also observed this for detritivores in our investigation (e.g., L. liba and A. delosa). Because C. clio was intensively infected, however, our study also provided evidence that predatory insects can be paratenic hosts. Coiled juveniles were typically present in muscle and fat body and present in almost all body regions. Not every infected paratenic host had external signs of infection; thus, dissections are required for accurate determination of infection prevalence and intensity. Our findings, in conjunction with those made in previous studies, indicate that a wide variety of aquatic insects may be used as paratenic hosts by Pheromermis. Because of their small size, it is highly likely that coiled juveniles are either overlooked or mistaken for melanized integumental wounds during field studies of aquatic insects. A more careful inspection for these coiled juveniles in aquatic insects, especially detritivores and their predators in seepage areas, would probably reveal that Pheromermis is far more common than presently documented. Copyright 1999 Academic Press. PMID- 10388552 TI - Hirsutellin A displays significant homology to microbial extracellular ribonucleases. PMID- 10388553 TI - Raman optical activity of filamentous bacteriophages: hydration of alpha-helices. AB - We report the first observations of vibrational Raman optical activity (ROA) on intact viruses. Specifically, ROA spectra of the filamentous bacteriophages Pf1, M13 and IKe in aqueous solution were measured in the range approximately 600-1800 cm-1. On account of its ability to probe directly the chiral elements of biomolecular structure, ROA has provided a new perspective on the solution structures of these well-studied systems. The ROA spectra of all three are dominated by signatures of helical elements in the major coat proteins, as expected from pre-existing data. The helical elements generate strong sharp positive ROA bands at approximately 1300 and 1342 cm-1in H2O solution, but in2H2O solution the approximately 1342 cm-1bands disappear completely. The spectra are similar to those of polypeptides under conditions that produce alpha-helical conformations. Our present results, together with results from other studies, suggest that the positive approximately 1342 cm-1ROA bands are generated by a highly hydrated form of alpha-helix, and that the positive approximately 1300 cm 1bands originate in alpha-helix in a more hydrophobic environment. The presence of significant amounts of highly hydrated helical sequences accords with the known flexibility of these viruses. Differences of spectral detail for Pf1, M13 and IKe demonstrate that ROA is sensitive to subtle variations of conformation and hydration within the major coat proteins, with M13 and IKe possibly containing more non-helical structure than Pf1. The ROA spectra of Pf1 at temperatures above and below that at which a structural transition is known to occur (approximately 10 degrees C) reveal little difference in the protein conformation between the two forms, but there are indications of changes in DNA structure. PMID- 10388554 TI - Display of epitopes on the surface of tobacco mosaic virus: impact of charge and isoelectric point of the epitope on virus-host interactions. AB - The biophysical properties of the tobacco mosaic tobamovirus (TMV) coat protein (CP) make it possible to display foreign peptides on the surface of TMV. The immunogenic epitopes G5-24 from the rabies virus (RV) glycoprotein, and 5B19 from murine hepatitis virus (MHV) S-glycoprotein were successfully displayed on the surface of TMV, and viruses accumulated to high levels in infected leaves of Nicotiana tabacum Xanthi-nn. The peptide RB19, which contains an arginine residue plus the 5B19 epitope fused to the CP (TMV-RB19), resulted in the induction of necrotic local lesions on inoculated leaves of N. tabacum Xanthi-nn and cell death of infected BY2 protoplasts. RNA dot blot assays confirmed that expression of the acidic and basic pathogenesis-related PR2 genes were induced in infected Xanthi-nn leaf tissue. TMV that carried epitope 31D from the RV nucleoprotein did not accumulate in inoculated tobacco leaves. Analysis of hybrid CPs predicted that the isoelectric points (pI):charge value was 5.31:-2 for wild-type CP, 5.64: 1 for CP-RB19, and 9.14:+2 for CP-31D. When acidic amino acids were inserted in CP-RB19 and CP-31D to bring their pI:charge to near that of wild-type CP, the resulting viruses TMV-RB19E and TMV-4D:31D infected N. tabacum Xanthi-nn plants and BY2 protoplasts without causing cell death. These data show the importance of the pI of the epitope and its effects on the hybrid CP pI:charge value for successful epitope display as well as the lack of tolerance to positively charged epitopes on the surface of TMV. PMID- 10388556 TI - DNA-induced conformational changes in cyclic AMP receptor protein: detection and mapping by a protein footprinting technique using multiple chemical proteases. AB - Cyclic AMP receptor protein (CRP) is a regulator of transcription in Escherichia coli which mediates its activity by binding specific DNA sequences in a cyclic AMP-dependent manner. The interaction of CRP with specific DNA was probed by a protein footprinting technique using chemical proteases of different charge, size, and hydrophobicity. The experimental data were compared with known crystal structures of cAMP-CRP and cAMP-CRP-DNA complexes to determine a correlation between the structure of the complexes, the nature of the chemical protease and protein cleavage patterns. In addition, such comparison allowed us to determine if DNA binding in solution induced conformational changes in the protein not apparent in the crystal structure. In the cAMP-CRP-DNA complex, both the protections and the enhancements of proteolytic cleavage were observed outside of the known CRP-DNA interface, suggesting that CRP undergoes a conformational change upon binding DNA. Among the observed changes, the most interesting were those around the B alpha-helix and beta-strand 8, since this region overlaps with the activation region 2 which CRP uses for protein-protein interactions with RNA polymerase. DNA-induced changes were observed also in the region involved in CRP CytR interaction and in CRP intersubunit contact regions. These data suggest that binding of DNA in solution induces conformational changes in CRP which can be transmitted via intersubunit contacts to regions of the protein involved in interactions with other members of transcriptional machinery. PMID- 10388555 TI - Induction of activator protein 1 (AP-1) in macrophages by human immunodeficiency virus type-1 NEF is a cell-type-specific response that requires both hck and MAPK signaling events. AB - Human immunodeficiency virus type 1 (HIV-1) Nef is important for viral infectivity and pathogenicity. HIV-1 infection is associated with inappropriate activation and defects in the function of monocytes/macrophages. We have studied the effects of HIV-1 Nef in the murine (RAW264.7) and human (THP-1) monocyte macrophage cell lines. Investigation of the activator protein-1 (AP-1) transcription factor showed that Nef expression induced both its DNA binding and transcriptional activities. Increased AP-1 DNA binding activity in RAW264.7 cells was associated with raised levels of c-Fos expression and induction of mRNA for the AP-1 responsive tissue inhibitor of metalloproteinases-1 (TIMP-1) gene. Mutagenesis and kinase inhibition studies were employed to determine signaling pathways used by Nef to induce AP-1. Data from these studies indicated that induction of AP-1 by Nef is likely to be mediated through the MAPK (ERK1 and 2) signaling pathway and requires the proline-rich PxxP motif of Nef, suggesting the involvement of upstream protein kinases belonging to the Src family. Effects of Nef on AP-1 induction were cell lineage-specific, being stimulatory in macrophages, inhibitory in T cells and without effect in HeLa cells. These latter two observations led us to test the possibility that cell-specific interactions of Nef with Src family proteins may modulate AP-1 activity. To this end we demonstrated that a dominant-negative Hck mutant caused inhibition of Nef mediated AP-1 DNA binding activity in RAW cells. In conclusion, induction of AP-1 by Nef is a specific feature of human and murine macrophage cell lines that requires signal transduction events involving Hck and MAPKs. PMID- 10388557 TI - The McrBC endonuclease translocates DNA in a reaction dependent on GTP hydrolysis. AB - McrBC specifically recognizes and cleaves methylated DNA in a reaction dependent on GTP hydrolysis. DNA cleavage requires at least two recognition sites that are optimally separated by 40-80 bp, but can be spaced as far as 3 kb apart. The nature of the communication between two recognition sites was analyzed on DNA substrates containing one or two recognition sites. DNA cleavage of circular DNA required only one methylated recognition site, whereas the linearized form of this substrate was not cleaved. However, the linearized substrate was cleaved if a Lac repressor was bound adjacent to the recognition site. These results suggest a model in which communication between two remote sites is accomplished by DNA translocation rather than looping. A mutant protein with defective GTPase activity cleaved substrates with closely spaced recognition sites, but not substrates where the sites were further apart. This indicates that McrBC translocates DNA in a reaction dependent on GTP hydrolysis. We suggest that DNA cleavage occurs by the encounter of two DNA-translocating McrBC complexes, or can be triggered by non-specific physical obstacles like the Lac repressor bound on the enzyme's path along DNA. Our results indicate that McrBC belongs to the general class of DNA "motor proteins", which use the free energy associated with nucleoside 5'-triphosphate hydrolysis to translocate along DNA. PMID- 10388558 TI - Comparative sequence analysis of the complete human sarcomeric myosin heavy chain family: implications for functional diversity. AB - The conventional myosin motor proteins that drive mammalian skeletal and cardiac muscle contraction include eight sarcomeric myosin heavy chain (MyHC) isoforms. Six skeletal MyHCs are encoded by genes found in tightly linked clusters on human and mouse chromosomes 17 and 11, respectively. The full coding regions of only two out of six mammalian skeletal MyHCs had been sequenced prior to this work. In an effort to assess the extent of sequence diversity within the human MyHC family we present new full-length coding sequences corresponding to four additional human genes: MyHC-IIb, MyHC-extraocular, MyHC-IIa and MyHC-IIx/d. This represents the first opportunity to compare the full coding sequences of all eight sarcomeric MyHC isoforms within a vertebrate organism. Sequence variability has been analyzed in the context of available structure/function data with an emphasis on potential functional diversity within the family. Results indicate that functional diversity among MyHCs is likely to be accomplished by having small pockets of sequence diversity in an otherwise highly conserved molecule. PMID- 10388559 TI - Time-resolved equatorial X-ray diffraction studies of skinned muscle fibres during stretch and release. AB - Equatorial X-ray diffraction patterns were recorded from small bundles of one to three chemically skinned frog sartorius muscle fibres (time resolution 250 microseconds) during rapid stretch and subsequent release. In the relaxed state, the dynamic A-band lattice spacing change as a result of a 2 % step stretch (determined from the positions of the 10 and 11 reflections) resulted in a 21 % increase in lattice volume, while static studies of spacing and sarcomere length indicated than an increase in volume of >/=50 % for the same length change. In rigor, stretch caused a lattice volume decrease which was reversed by a subsequent release. In activated fibres (pCa 4.5) exposed to 10 mM 2,3 butanedione 2-monoxime (BDM), stretch was accompanied by a lattice compression exceeding that of constant volume behaviour, but during tension recovery, compression was partially reversed to leave a net spacing change close to that observed in the relaxed fibre. In the relaxed state, spacing changes were correlated with the amplitude of the length step, while in rigor and BDM states, spacing changes correlated more closely with axial force. This behaviour is explicable in terms of two components of radial force, one due to structural constraints as seen in the relaxed state, and an additional component arising from cross-bridge formation. The ratio of axial to radial force for a single thick filament resulting from a length step was four in rigor and BDM, but close to unity for the relaxed state. PMID- 10388560 TI - An amphipathic alpha-helix at a membrane interface: a structural study using a novel X-ray diffraction method. AB - The amphipathic alpha-helix is a recurrent feature of membrane-active proteins, peptides, and toxins. Despite extensive biophysical studies, the structural details of its affinity for membrane interfaces remain rather vague. We report here the first results of an effort to obtain detailed structural information about alpha-helices in membranes by means of a novel X-ray diffraction method. Specifically, we determined the transbilayer position and orientation of an archetypal class A amphipathic helical peptide in oriented fluid-state dioleoylphosphatidylcholine (DOPC) bilayers. The peptide, Ac-18A-NH2(Ac DWLKAFYDKVAEKLKEAF-NH2), is a model for class A amphipathic helices of apolipoprotein A-I and other exchangeable lipoproteins. The diffraction method relies upon experimental determinations of absolute scattering-length density profiles along the bilayer normal and the transbilayer distribution of the DOPC double bonds by means of specific bromination, and molecular modeling of the perturbed lipid bilayer (derived using the transbilayer distribution of the double bonds) and the peptide. The diffraction results showed that Ac-18A-NH2was located in the bilayer interface and that its transbilayer distribution could be described by a Gaussian function with a 1/e-halfwidth of 4.5(+/-0.3) A located 17.1(+/-0.3) A from the bilayer center, close to the glycerol moiety. Molecular modeling suggested that Ac-18A-NH2is helical and oriented generally parallel with the bilayer plane. The helicity and orientation were confirmed by oriented circular dichroism measurements. The width of the Gaussian distribution, a measure of the diameter of the helix, indicated that the Ac-18A-NH2helix penetrated the hydrocarbon core to about the level of the DOPC double bonds. Bilayer perturbations caused by Ac-18A-NH2were surprisingly modest, consisting of a slight decrease in bilayer thickness with a concomitant shift of the double bond distribution toward the bilayer center, as expected from a small increase in lipid-specific area caused by the peptide. PMID- 10388561 TI - Structure and thermodynamics of metal binding in the P5 helix of a group I intron ribozyme. AB - The solution structure of an RNA hairpin modelling the P5 helix of a group I intron, complexed with Co(NH3)63+, has been determined by nuclear magnetic resonance. Co(NH3)63+, which possesses a geometry very close to Mg(H2O)62+, was used to identify and characterize a Mg2+binding site in the RNA. Strong and positive intermolecular nuclear Overhauser effect (NOE) cross-peaks define a specific complex in which the Co(NH3)63+molecule is in the major groove of tandem G.U base-pairs. The structure of the RNA is characterized by a very low twist angle between the two G.U base-pairs, providing a flat and narrowed major groove. The Co(NH3)63+, although highly localized, is free to rotate to hydrogen bond in several ways to the O4 atoms of the uracil bases and to N7 and O6 of the guanine bases. Negative and small NOE cross-peaks to other protons in the sequence reveal a non-specific or delocalized interaction, characterized by a high mobility of the cobalt ion. Mn2+titrations of P5 show specific broadening of protons of the G.U base-pairs that form the metal ion binding site, in agreement with the NOE data from Co(NH3)63+. Binding constants for the interaction of Co(NH3)63+and of Mg2+to P5 were determined by monitoring imino proton chemical shifts during titration of the RNA with the metal ions. Dissociation constants are on the order of 0.1 mM for Co(NH3)63+and 1 mM for Mg2+. Binding studies were done on mutants with sequences corresponding to the three orientations of tandem G.U base-pairs. The affinities of Co(NH3)63+and Mg2+for the tandem G.U base-pairs depend strongly on their sequences; the differences can be understood in terms of the different structures of the corresponding metal ion-RNA complexes. Substitution of G.C or A.U for G.U pairs also affected the binding, as expected. These structural and thermodynamic results provide systematic new information about major groove metal ion binding in RNA. PMID- 10388562 TI - DNA binding mediates conformational changes and metal ion coordination in the active site of PcrA helicase. AB - Based upon the crystal structures of PcrA helicase, we have made and characterised mutations in a number of conserved helicase signature motifs around the ATPase active site. We have also determined structures of complexes of wild type PcrA with ADPNP and of a mutant PcrA complexed with ADPNP and Mn2+. The kinetic and structural data define roles for a number of different residues in and around the ATP binding site. More importantly, our results also show that there are two functionally distinct conformations of ATP in the active site. In one conformation, ATP is hydrolysed poorly whereas in the other (activated) conformation, ATP is hydrolysed much more rapidly. We propose a mechanism to explain how the stimulation of ATPase activity afforded by binding of single stranded DNA stabilises the activated conformation favouring Mg2+binding and a consequent repositioning of the gamma-phosphate group which promotes ATP hydrolysis. A part of the associated conformational change in the protein forces the side-chain of K37 to vacate the Mg2+binding site, allowing the cation to bind and interact with ATP. PMID- 10388563 TI - Crystal structures of the neurotrophin-binding domain of TrkA, TrkB and TrkC. AB - The Trk receptors and their neurotrophin ligands control development and maintenance of the nervous system. The crystal structures of the ligand binding domain of TrkA, TrkB, and TrkC were solved and refined to high resolution. The domains adopt an immunoglobulin-like fold, but crystallized in all three instances as dimers with the N-terminal strand of each molecule replaced by the same strand of a symmetry-related mate. Models of the correctly folded domains could be constructed by changing the position of a single residue, and the resulting model of the binding domain of TrkA is essentially identical with the bound structure as observed in a complex with nerve growth factor. An analysis of the existing mutagenesis data for TrkA and TrkC in light of these structures reveals the structural reasons for the specificity among the Trk receptors, and explains the underpinnings of the multi-functional ligands that have been reported. The overall structure of all three domains belongs to the I-set of immunoglobulin-like domains, but shows several unusual features, such as an exposed disulfide bridge linking two neighboring strands in the same beta-sheet. For all three domains, the residues that deviate from the standard fingerprint pattern common to the I-set family fall in the region of the ligand binding site observed in the complex. Therefore, identification of these deviations in the sequences of other immunoglobulin-like domain-containing receptors may help to identify their ligand binding site even in the absence of structural or mutagenesis data. PMID- 10388564 TI - Apo and holo crystal structures of an NADP-dependent aldehyde dehydrogenase from Streptococcus mutans. AB - The aldehyde dehydrogenases (ALDHs) are a superfamily of multimeric enzymes which catalyse the oxidation of a broad range of aldehydes into their corresponding carboxylic acids with the reduction of their cofactor, NAD or NADP, into NADH or NADPH. At present, the only known structures concern NAD-dependent ALDHs. Three structures are available in the Protein Data Bank: two are tetrameric and the other is a dimer. We solved by molecular replacement the first structure of an NADP-dependent ALDH isolated from Streptococcus mutans, in its apo form and holo form in complex with NADP, at 1.8 and 2.6 A resolution, respectively. Although the protein sequence shares only approximately 30 % identity with the other solved tetrameric ALDHs, the structures are very similar. However, a large local conformational change in the region surrounding the 2' phosphate group of the adenosine moiety is observed when the enzyme binds NADP, in contrast to the NAD dependent ALDHs. Structure and sequence analyses reveal several properties. A small number of residues seem to determine the oligomeric state. Likewise, the nature (charge and volume) of the residue at position 180 (Thr in ALDH from S. mutans) determines the cofactor specificity in comparison with the structures of NAD-dependent ALDHs. The presence of a hydrogen bond network around the cofactor not only allows it to bind to the enzyme but also directs the side-chains in a correct orientation for the catalytic reaction to take place. Moreover, a specific part of this network appears to be important in substrate binding. Since the enzyme oxidises the same substrate, glyceraldehyde-3-phosphate (G3P), as NAD dependent phosphorylating glyceraldehyde-3-phosphate dehydrogenases (GAPDH), the active site of GAPDH was compared with that of the S. mutans ALDH. It was found that Arg103, Arg283 and Asp440 might be key residues for substrate binding. PMID- 10388565 TI - Crystal structure of neurotoxin Ts1 from Tityus serrulatus provides insights into the specificity and toxicity of scorpion toxins. AB - The crystal structure of neurotoxin Ts1, a major component of the venom of the Brazilian scorpion Tityus serrulatus, has been determined at 1.7 A resolution. It is the first X-ray structure of a highly toxic anti-mammalian beta-toxin. The folding of the polypeptide chain of Ts1 is similar to that of other scorpion toxins. A cysteine-stabilised alpha-helix/beta-sheet motif forms the core of the flattened molecule. All residues identified as functionally important by chemical modification and site-directed mutagenesis are located on one side of the molecule, which is therefore considered as the Na+channel recognition site. The distribution of charged and non-polar residues over this surface determines the specificity of the toxin-channel interaction. Comparison to other scorpion toxins shows that positively charged groups at positions 1 and 12 as well as a negative charge at position 2 are likely determinants of the specificity of beta-toxins. In contrast, the contribution of the conserved aromatic cluster to the interaction might be relatively small. Comparison of Ts1 to weak beta-toxins from Centruroides sculpturatus Ewing reveals that a number of basic amino acid residues located on the face of the molecule opposite to the binding surface may account for the high toxicity of Ts1. PMID- 10388566 TI - Insights into carbohydrate recognition by Narcissus pseudonarcissus lectin: the crystal structure at 2 A resolution in complex with alpha1-3 mannobiose. AB - Carbohydrate recognition by monocot mannose-binding lectins was studied via the crystal structure determination of daffodil (Narcissus pseudonarcissus) lectin. The lectin was extracted from daffodil bulbs, and crystallised in the presence of alpha-1,3 mannobiose. Molecular replacement methods were used to solve the structure using the partially refined model of Hippeastrum hybrid agglutinin as a search model. The structure was refined at 2.0 A resolution to a final R -factor of 18.7 %, and Rfreeof 26.7 %. The main feature of the daffodil lectin structure is the presence of three fully occupied binding pockets per monomer, arranged around the faces of a triangular beta-prism motif. The pockets have identical topology, and can bind mono-, di- or oligosaccharides. Strand exchange forms tightly bound dimers, and higher aggregation states are achieved through hydrophobic patches on the surface, completing a tetramer with internal 222 symmetry. There are therefore 12 fully occupied binding pockets per tetrameric cluster. The tetramer persists in solution, as shown with small-angle X-ray solution scattering. Extensive sideways and out-of-plane interactions between tetramers, some mediated via the ligand, make up the bulk of the lattice contacts.A fourth binding site was also observed. This is unique and has not been observed in similar structures. The site is only partially occupied by a ligand molecule due to the much lower binding affinity. A comparison with the Galanthus nivalis agglutinin/mannopentaose complex suggests an involvement of this site in the recognition mechanism for naturally occurring glycans. PMID- 10388567 TI - Crystal structure of a mammalian purple acid phosphatase. AB - Tartrate-resistant acid phosphatase (TRAP) is a mammalian di-iron- containing enzyme that belongs to the family of purple acid phosphatases (PAP). It is highly expressed in a limited number of tissues, predominantly in bone-resorbing osteoclasts and in macrophages of spleen. We have determined the crystal structure of rat TRAP in complex with a phosphate ion to 2.7 A resolution. The fold resembles that of the catalytic domain of kidney bean purple acid phosphatase (KBPAP), although the sequence similarity is limited to the active site residues. A surface loop near the active site is absent due to proteolysis, leaving the active-site easily accessible from the surrounding solvent. This, we believe, gives a structural explanation for the observed proteolytic activation of TRAP. The current structure was determined at a relatively high pH and without any external reducing agents. It is likely that it represents an oxidized and therefore catalytically inactive form of the enzyme. PMID- 10388568 TI - Characterization of the binding interface between ubiquitin and class I human ubiquitin-conjugating enzyme 2b by multidimensional heteronuclear NMR spectroscopy in solution. AB - Ubiquitin-conjugating enzymes (Ubc) are involved in ubiquitination of proteins in the protein degradation pathway of eukaryotic cells. Ubc transfers the ubiquitin (Ub) molecules to target proteins by forming a thioester bond between their active site cysteine residue and the C-terminal glycine residue of ubiquitin. Here, we report on the NMR assignment and secondary structure of class I human ubiquitin-conjugating enzyme 2b (HsUbc2b). Chemical shift perturbation studies allowed us to map the contact area and binding interface between ubiquitin and HsUbc2b by1H-15N HSQC NMR spectroscopy. The serine mutant of the active site Cys88 of HsUbc2b was employed to obtain a relatively stable covalent ubiquitin complex of HsUbc2b(C88S). Changes in chemical shifts of amide protons and nitrogen atoms induced by the formation of the covalent complex were measured by preparing two segmentally labeled complexes with either ubiquitin or HsUbc2b(C88S)15N-labeled. In ubiquitin, the interaction is primarily sensed by the C-terminal segment Val70 - Gly76, and residues Lys48 and Gln49. The surface area on ubiquitin, as defined by these residues, overlaps partially with the presumed binding site with ubiquitin-activating enzyme (E1). In HsUbc2b, most of the affected residues cluster in the vicinity of the active site, namely, around the active site Cys88 itself, the second alpha-helix, and the flexible loop which connects helices alpha2 and alpha3 and which is adjacent to the active site. An additional site on HsUbc2b for a weak interaction with ubiquitin could be detected in a titration study where the two proteins were not covalently linked. This site is located on the backside of HsUbc2b opposite to the active site and is part of the beta-sheet. The covalent and non-covalent interaction sites are clearly separated on the HsUbc2b surface, while no such clear-cut segregation of the interaction area was observed on ubiquitin. PMID- 10388569 TI - Domain motions accompanying Tet repressor induction defined by changes of interspin distances at selectively labeled sites. AB - To investigate internal movements in Tet repressor (TetR) during induction by tetracycline (tc) we determined the interspin distances between pairs of nitroxide spin labels attached to specific sites by electron paramagnetic resonance (EPR) spectroscopy. For this purpose, we constructed six TetR variants with engineered cysteine pairs located in regions with presumed conformational changes. These are I22C and N47C in the DNA reading head, T152C/Q175C, A161C/Q175C and R128C/D180C near the tc-binding pocket, and T202C in the dimerization surface. All TetR mutants show wild-type activities in vivo and in vitro. The binding of tc results in a considerable decrease of the distance between the nitroxide groups attached to both I22C residues in the TetR dimer and an increase of the distance between the N47C residues. These opposite effects are consistent with a twisting motion of the DNA reading heads. Changes of the spin spin interactions between nitroxide groups attached to residues near the tc binding pocket demonstrate that the C-terminal end of alpha-helix 9 moves away from the protein core upon DNA binding. Alterations of the dipolar interaction between nitroxide groups at T202C indicate different conformations for tc and DNA bound repressor also in the dimerization area. These results are used to model structural changes of TetR upon induction. PMID- 10388571 TI - The packing density in proteins: standard radii and volumes. AB - The sizes of atomic groups are a fundamental aspect of protein structure. They are usually expressed in terms of standard sets of radii for atomic groups and of volumes for both these groups and whole residues. Atomic groups, which subsume a heavy-atom and its covalently attached hydrogen atoms into one moiety, are used because the positions of hydrogen atoms in protein structures are generally not known. We have calculated new values for the radii of atomic groups and for the volumes of atomic groups. These values should prove useful in the analysis of protein packing, protein recognition and ligand design. Our radii for atomic groups were derived from intermolecular distance calculations on a large number (approximately 30,000) of crystal structures of small organic compounds that contain the same atomic groups to those found in proteins. Our radii show significant differences to previously reported values. We also use this new radii set to determine the packing efficiency in different regions of the protein interior. This analysis shows that, if the surface water molecules are included in the calculations, the overall packing efficiency throughout the protein interior is high and fairly uniform. However, if the water structure is removed, the packing efficiency in peripheral regions of the protein interior is underestimated, by approximately 3.5 %. PMID- 10388570 TI - A single tyrosine prevents insertion of ribonucleotides in the eukaryotic-type phi29 DNA polymerase. AB - Three conserved motifs (named A, B and C) have been proposed to form the polymerization active site in all classes of DNA-dependent polymerases. In eukaryotic-type (alpha-like) DNA polymerases, motif A is characterized by the consensus "Dx2SLYP". Mutants in phi29 DNA polymerase residue Tyr254 of this conserved motif had been previously shown to be affected in dNTP binding. Here, we show that a single substitution of Tyr254 into a valine residue enables the enzyme to incorporate ribonucleotide substrates, without affecting its wild-type affinity for dNTPs. Whereas the wild-type enzyme preferred dNTPs more than two million-fold over rNTPs, the mutation of Tyr254 into valine reduced the discrimination for rNTPs up to 1000-fold. In addition to this discrimination mechanism, based on sugar selection, phi29 DNA polymerase is very inefficient when extending an RNA primer terminus, allowing its exonucleolytic degradation. These results indicate that the Tyr254 of phi29 DNA polymerase is responsible for the discrimination against the 2'-OH group of an incoming ribonucleotide. This is the first time that the invariant tyrosine residue of motif A is involved in ribo versus deoxyribonucleotide discrimination in an eukaryotic-type DNA polymerase. PMID- 10388572 TI - Ab initio fold prediction of small helical proteins using distance geometry and knowledge-based scoring functions. AB - The problem of protein tertiary structure prediction from primary sequence can be separated into two subproblems: generation of a library of possible folds and specification of a best fold given the library. A distance geometry procedure based on random pairwise metrization with good sampling properties was used to generate a library of 500 possible structures for each of 11 small helical proteins. The input to distance geometry consisted of sets of restraints to enforce predicted helical secondary structure and a generic range of 5 to 11 A between predicted contact residues on all pairs of helices. For each of the 11 targets, the resulting library contained structures with low RMSD versus the native structure. Near-native sampling was enhanced by at least three orders of magnitude compared to a random sampling of compact folds. All library members were scored with a combination of an all-atom distance-dependent function, a residue pair-potential, and a hydrophobicity function. In six of the 11 cases, the best-ranking fold was considered to be near native. Each library was also reduced to a final ab initio prediction via consensus distance geometry performed over the 50 best-ranking structures from the full set of 500. The consensus results were of generally higher quality, yielding six predictions within 6.5 A of the native fold. These favorable predictions corresponded to those for which the correlation between the RMSD and the scoring function were highest. The advantage of the reported methodology is its extreme simplicity and potential for including other types of structural restraints. PMID- 10388573 TI - Analysis methods for comparison of multiple molecular dynamics trajectories: applications to protein unfolding pathways and denatured ensembles. AB - In molecular dynamics simulations of protein unfolding, the pathway of one protein molecule is studied at a time. In contrast, experimental denaturation studies sample from large ensembles of molecules passing from the native to unfolded state. If reasonable comparisons with experiment are to be made, then the generality of the simulations needs to be confirmed by performing multiple unfolding simulations. Given that protein unfolding trajectories are very complicated functions of the proteins and the environment, comparing different trajectories, even under the same conditions, is not straightforward. Several methods are presented here that attempt to accomplish this task at different levels of complexity. The simpler methods are geometry based and make use of the root-mean-squared deviations between structures, while the more complicated methods are based on the time variation of the various properties of the system during the unfolding process. These methods are applied to multiple simulations of three different proteins, bovine pancreatic trypsin inhibitor, chymotrypsin inhibitor 2, and barnase. In general, for these three proteins protein unfolding proceeded via expansion of the core and fraying of secondary structure to yield the major transition state. Once past the transition state, the trajectories for a given protein diverged as the protein lost further secondary and tertiary structure by a variety of mechanisms. Although the unfolding pathways diverged, similar conformations were populated in the denatured state even when the unfolding occurred via different pathways. The multitude of different pathways leading to the denatured state agrees with the funnel description of protein folding. Although the pathways differed in conformational space, the physical properties of the conformations were often similar, highlighting the danger of assuming that similar observed properties imply similar conformations. In fact, there may be many different "conformational pathways" of unfolding that fit within a preferred "property space pathway". PMID- 10388574 TI - Side-chain and backbone flexibility in protein core design. AB - We have developed a computational approach for the design and prediction of hydrophobic cores that includes explicit backbone flexibility. The program consists of a two-stage combination of a genetic algorithm and monte carlo sampling using a torsional model of the protein. Backbone structures are evaluated either by a canonical force-field or a constraining potential that emphasizes the preservation of local geometry. The utility of the method for protein design and engineering is explored by designing three novel hydrophobic core variants of the protein 434 cro. We use the new method to evaluate these and previously designed 434 cro variants, as well as a series of phage T4 lysozyme variants. In order to properly evaluate the influence of backbone flexibility, we have also analyzed the effects of varying amounts of side-chain flexibility on the performance of fixed backbone methods. Comparison of results using a fixed versus flexible backbone reveals that, surprisingly, the two methods are almost equivalent in their abilities to predict relative experimental stabilities, but only when full side-chain flexibility is allowed. The prediction of core side chain structure can vary dramatically between methods. In some, but not all, cases the flexible backbone method is a better predictor of structure. The development of a flexible backbone approach to core design is particularly important for attempts at de novo protein design, where there is no prior knowledge of a precise backbone structure. PMID- 10388575 TI - Energetics of a hydrogen bond (charged and neutral) and of a cation-pi interaction in apoflavodoxin. AB - Anabaena apoflavodoxin contains a single histidine residue (H34) that interacts with two aromatic residues (F7 and Y47). The histidine and phenylalanine rings are almost coplanar and they can establish a cation-pi interaction when the histidine is protonated. The histidine and tyrosine side-chains are engaged in a hydrogen bond, which is their only contact. We analyse the energetics of these interactions using p Ka-shift analysis, double-mutant cycle analysis at two pH values, and X-ray crystallography. The H/F interaction is very weak when the histidine is neutral, but it is strengthened by 0.5 kcal mol-1on histidine protonation. Supporting this fact, the histidine p Kain a F7L mutant is 0.4 pH units lower than in wild-type. The strength of the H/Y hydrogen bond is 0.7 kcal mol-1when the histidine is charged, and it becomes stronger (1.3 kcal mol-1) when the histidine is neutral. This is consistent with our observation that the (H34)Nepsilon2-OH(Y47) distance is slightly shorter in the apoflavodoxin structure at pH 9.0 than in the previously reported structure at pH 6.0. It is also consistent with a histidine p Kavalue 0.6 pH units higher in a Y47F mutant than in the wild-type protein. We suggest that the higher stability of the neutral hydrogen bond could be due to a higher desolvation penalty of the charged hydrogen bond that would offset its more favourable enthalpy of formation. The relationship between hydrogen bond strength and the contribution of hydrogen bonds to protein stability is discussed. PMID- 10388576 TI - Equilibrium folding properties of the yeast prion protein determinant Ure2. AB - The yeast non-Mendelian factor [URE3] propagates by a prion-like mechanism, involving aggregation of the chromosomally encoded protein Ure2. The [URE3] phenotype is equivalent to loss of function of Ure2, a protein involved in regulation of nitrogen metabolism. The prion-like behaviour of Ure2 in vivo is dependent on the first 65 amino acid residues of its N-terminal region which contains a highly repetitive sequence rich in asparagine. This region has been termed the prion-determining domain (PrD). Removal of as little as residues 2-20 of the protein is sufficient to prevent occurrence of the [URE3] phenotype. Removal of the PrD does not affect the regulatory activity of Ure2. The C terminal portion of the protein has homology to glutathione S -transferases, which are dimeric proteins. We have produced the Ure2 protein to high yield in Escherichia coli from a synthetic gene. The recombinant purified protein is shown to be a dimer. The stability, folding and oligomeric state of Ure2 and a series of N-terminally truncated or deleted variants were studied and compared. The stability of Ure2, DeltaGD-N, H2O, determined by chemical denaturation and monitored by fluorescence, is 12.1(+/-0.4) kcal mol-1at 25 degrees C and pH 8.4. A range of structural probes show a single, coincident unfolding transition, which is invariant over a 550-fold change in protein concentration. The stability is the same within error for Ure2 variants lacking all or part of the prion determining domain. The data indicate that in the folded protein the PrD is in an unstructured conformation and does not form specific intra- or intermolecular interactions at micromolar protein concentrations. This suggests that the C terminal domain may stabilise the PrD against prion formation by steric means, and implies that the PrD does not induce prion formation by altering the thermodynamic stability of the folded protein. PMID- 10388577 TI - Novel molecular architecture of the multimeric archaeal PEP-synthase homologue (MAPS) from Staphylothermus marinus. AB - The phosphoenolpyruvate (PEP)-synthases belong to the family of structurally and functionally related PEP-utilizing enzymes. The only archaeal member of this family characterized thus far is the Multimeric Archaeal PEP-Synthase homologue from Staphylothermus marinus (MAPS). This protein complex differs from the bacterial and eukaryotic representatives characterized to date in its homomultimeric, as opposed to dimeric or tetrameric, structure. We have probed the molecular architecture of MAPS using limited proteolytic digestion in conjunction with electron microscopic, biochemical, and biophysical techniques. The 2.2 MDa particle was found to be organized in a concentric fashion. The 93.7 kDa monomers possess a pronounced tripartite domain structure and are arranged such that the N-terminal domains form an outer shell, the intermediate domains form an inner shell, and the C-terminal domains form a core structure responsible for the assembly into a multimeric complex. The core domain was shown to be capable of assembling into the native multimer by recombinant expression in Escherichia coli. Deletion mutants as well as a synthetic peptide were investigated for their state of oligomerization using native polyacrylamide gel electrophoresis, molecular sieve chromatography, analytical ultracentrifugation, circular dichroism (CD) spectroscopy, and chemical cross-linking. Our data confirmed the existence of a short C-terminal, alpha-helical oligomerization motif that had been suggested by multiple sequence alignments and secondary structure predictions. We propose that this motif bundles the monomers into six groups of four. An additional formation of 12 dimers between globular domains from different bundles leads to the multimeric assembly. According to our model, each of the six bundles of globular domains is positioned at the corners of an imaginary octahedron, and the helical C-terminal segments are oriented towards the centre of the particle. The edges of the octahedron represent the dimeric contacts. Phylogenetic analysis suggests that the ancient predecessor of this family of enzymes contained the C-terminal oligomerization motif as a feature that was preserved in some hyperthermophiles. PMID- 10388578 TI - Comparison of scalar measures used in magnetic resonance diffusion tensor imaging. AB - The tensors derived from diffusion tensor imaging describe complex diffusion in tissues. However, it is difficult to compare tensors directly or to produce images that contain all of the information of the tensor. Therefore, it is convenient to produce scalar measures that extract desired aspects of the tensor. These measures map the three-dimensional eigenvalues of the diffusion tensor into scalar values. The measures impose an order on eigenvalue space. Many invariant scalar measures have been introduced in the literature. In the present manuscript, a general approach for producing invariant scalar measures is introduced. Because it is often difficult to determine in clinical practice which of the many measures is best to apply to a given situation, two formalisms are introduced for the presentation, definition, and comparison of measures applied to eigenvalues: (1) normalized eigenvalue space, and (2) parametric eigenvalue transformation plots. All of the anisotropy information contained in the three eigenvalues can be retained and displayed in a two-dimensional plot, the normalized eigenvalue plot. An example is given of how to determine the best measure to use for a given situation by superimposing isometric contour lines from various anisotropy measures on plots of actual measured eigenvalue data points. Parametric eigenvalue transformation plots allow comparison of how different measures impose order on normalized eigenvalue space to determine whether the measures are equivalent and how the measures differ. These formalisms facilitate the comparison of scalar invariant measures for diffusion tensor imaging. Normalized eigenvalue space allows presentation of eigenvalue anisotropy information. PMID- 10388579 TI - A W-band pulsed ENDOR spectrometer: setup and application to transition metal centers. AB - The design and performance of a 95 GHz pulsed W-band EPR/ENDOR spectrometer is described with emphasis on the ENDOR part. Its unique feature is the easy and fast sample exchange at 4.2 K for frozen solution and single crystal samples. In addition, the microwave bridge power output is relatively high (maximum 267 mW), which allows the application of short microwave pulses. This increases the sensitivity in echo experiments because of the broader excitation bandwidth and the possibility of employing short pulse intervals, as long as the dead time does not increase significantly with the power. The spectrometer features two microwave and radiofrequency (0.1-220 MHz, 3 kW pulse power) channels and a 6 T superconducting magnet in a solenoid configuration. The magnet is equipped with cryogenic sweep coils providing a sweep range of +/-0. 4 and +/-0.2 T for a center field of 0-4 and 4-6 T, respectively. The spectrometer performance is demonstrated on Cu(II) centers in single crystals, a zeolite polycrystalline sample, and a protein frozen solution. PMID- 10388580 TI - Transforming NMR data despite missing points. AB - Some NMR experiments produce data with several of the initial points missing. The inverse discrete Fourier transform (IDFT) assumes these points are present so the data cannot be so transformed without artifact-ridden results. This problem is often particularly severe when projection imaging with free-induction decays (FIDs). This paper compares recent methods for obtaining a projection from incomplete data and elaborates on their strengths and limitations. One method is to write the transform that would take the desired projection to the truncated data set, and then solve the matrix equation by singular value decomposition. A second replaces the missing data with zeros, so that an IDFT produces a projection with unwanted artifacts. Then one solves the matrix equation that takes the desired projection to the artifact-ridden projection. A third uses the same artifact-ridden projection, but fits the region outside the bandwidth of the sample with as many sinusoidal functions as there are missing data. The coefficients of these functions are estimates of the missing data, and the projection is obtained by transforming the completed FID or subtracting the extrapolation of the fitted curve from the region containing the object. We show that when all three methods are applicable, they theoretically produce the same result. They differ by ease of implementation and possibly by computational errors. They give a result similar to that of the previous method that iteratively corrects the FID and projection after repeated IDFTs and DFTs. We find that one can obtain a projection despite missing a substantial number of data. PMID- 10388581 TI - An integrated head immobilization system and high-performance RF coil for fMRI of visual paradigms at 1.5 T. AB - A flexible quadrature radiofrequency coil that maximizes the signal-to-noise ratio over the field of view of the human brain has been integrated into a head immobilization and visor system for fMRI at 1.5 T. Head motion is reduced by the visor that incorporates a head clamp and a simple visual sighting system that provides feedback on head position. This system is demonstrated in serial images by correction of deliberate head motions. The sensitivity at the cortical surface of fMRI using blood oxygenation level dependent contrast is increased significantly above that of the commercial rigid volume RF coil under the same acquisition conditions. This improved performance is demonstrated using visual activation and eye movement paradigms. PMID- 10388582 TI - Experimental evaluation of a surface charge method for computing the induced magnetic field in trabecular bone. AB - The magnetic field induced in the pores of trabecular bone as a result of the susceptibility difference between bone and bone marrow was computed with the aid of magnetic surface charge models generated from images of trabecular bone specimens acquired at 78 and 63 microm resolution. The predicted field was compared with the values derived from 2D and 3D field maps obtained by echo offset imaging techniques and excellent agreement was found between the two methods. Finally, from the slopes of regression between the experimental and computed fields, the absolute susceptibility of bone was nondestructively determined as -11.0 x 10(-6) (MKS), which is in close agreement with a reported value of -11.3 x 10(-6) obtained with powdered bone by means of a spectroscopic susceptibility matching technique (J. A. Hopkins and F. W. Wehrli, Magn. Reson. Med. 37, 494-500 (1997)). PMID- 10388583 TI - The accuracy of distance measurements in solid-state NMR. AB - The accuracy with which distances can be measured using dipolar recoupling experiments in solid-state NMR is investigated. The relative precision of experiments in a three spin system versus an isolated spin pair is found to depend very strongly on the nature of the coupling Hamiltonian. The accuracy of distances measured in even the simplified three spin system is seen to be very poor for existing homonuclear recoupling Hamiltonians. This suggests that it would be difficult to exploit broadband homonuclear recoupling to measure geometrical information reliably in complex spin systems. These conclusions apply equally to both single-crystal studies and powder samples. In contrast, the presence of additional spins has marginal impact on the accuracy when the coupling Hamiltonians commute with each other, as in the case of heteronuclear recoupling. The possibility of creating such a Hamiltonian for homonuclear recoupling using a suitable rotor-synchronized pulse sequence is discussed. PMID- 10388584 TI - Transient magnetic resonance without RF pulses: fast field switching. AB - An unusual strategy for performing magnetic resonance experiments is demonstrated. Instead of employing conventional radiofrequency transmitter fields to perturb spin state populations away from equilibrium, as is the basis of most magnetic resonance spectrometers today, technological advances now make possible fast switching of the magnetic field orientation to achieve the same effect. This is demonstrated with an electron spin resonance experiment where the magnetic field is switched 90 degrees nonadiabatically with a dead time of a few tens of nanoseconds and an electron free induction decay observed. PMID- 10388585 TI - Selective imaging of biofilms in porous media by NMR relaxation. AB - Nuclear magnetic resonance imaging (NMRI) techniques were employed to identify and selectively image biological films (biofilm) growing in aqueous systems. Biofilms are shown to affect both the longitudinal (T1) and transverse (T2) NMR relaxation time values of proximal water hydrogens. Results are shown for biofilm growth experiments performed in a transparent parallel-plate reactor. A comparison of biofilm distributions by both NMR and optical imaging yielded general agreement for both an open-flow system and an idealized porous system (the reactor without and with packed glass beads, respectively). The selective imaging of biofilm by relaxation NMRI is dependent upon the resolution of relaxation times for the fluid phases, dynamic range, and signal-to-noise ratio. For open-flow systems, the use of a rapid and quantitative T2-sorted NMRI technique was preferred. For porous systems where T2 values are generally more similar, a T1-weighted technique was preferred. PMID- 10388586 TI - Signal intensities in FLASH-EPI-hybrid sequences. AB - Theoretical considerations on the signal-to-noise ratio (SNR) in FLASH-EPI-Hybrid imaging were published previously. The purpose of this work was to investigate in vivo the signal intensities in Hybrid images as a function of sequence specific parameters. In detail, the SNR as a function of the number of echoes m per RF excitation, the excitation flip angle alpha, and the dependence on the tissue relaxation times T1 and T2* were studied. In eight healthy subjects brain and abdominal Hybrid images were acquired where m and alpha were changed independently. Signal intensities in human brain, liver, and kidney were evaluated for each Hybrid experiment. Additionally, T1 and T2* values of these tissue types were quantified to allow for a comparison with the theory. An excellent agreement between calculated and measured signal behavior was found. The theory was therefore validated in vivo and can thus be used to optimize the signal-to-noise in Hybrid experiments. PMID- 10388587 TI - An "openable," high-strength gradient set for orthopedic MRI. AB - A novel three-axis gradient set and RF resonator for orthopedic MRI has been designed and constructed. The set is openable and may be wrapped around injured joints. The design methodology used was the minimization of magnetic field spherical harmonics by simulated annealing. Splitting of the longitudinal coil presents the major design challenge to a fully openable gradient set and in order to efficiently design such coils, we have developed a new fast algorithm for determining the magnetic field spherical harmonics generated by an arc of multiturn wire. The algorithm allows a realistic impression of the effect of split longitudinal designs. A prototype set was constructed based on the new designs and tested in a 2-T clinical research system. The set generated 12 mT/m/A with a linear region of 12 cm and a switching time of 100 micros, conforming closely with theoretical predictions. Preliminary images from the set are presented. PMID- 10388588 TI - A novel high-gradient permanent magnet for the profiling of planar films and coatings. AB - The design and construction of a low-cost, permanent magnet is described. The magnet is intended for applications which require a large static gradient, such as those for which stray field imaging or fringe field diffusometry are conventionally employed. The magnet has been designed using the scalar potential method. Particular features of the magnet include a field profile such that ||B || is constant in the horizontal plane and such that B is horizontal at the midpoint between the poles. There is a vertical, and therefore orthogonal, strong gradient, G, in ||B ||. The ratio G/ ||B || is constant within a large volume and so allows measurements at a range of gradient strengths. It is this ratio which governs the shape of the pole-pieces. The constructed magnet has a typical operating field of 0.8 T, gives a gradient of 20 Tm-1, and has a useable interpole access of 20 mm. Field plot data show values consistent with the theory. In particular ||B || has a curvature of less than +/-5 microm over a 5 x 5 mm area at the target field. The magnet is most suitable for the one dimensional profiling of thin planar samples. As an example of the magnet's use, a profile of a sandwich structure made of several polymer layers is shown. In addition, a set of one-dimensional profiles of an alkyd coating, recorded during solvent loss and cross-linking, is presented. This example demonstrates quantitative T2 measurements at a resolution of 6.5 microm across a 70-microm thick film. PMID- 10388589 TI - Examples of Hartmann-Hahn match conditions for CP/MAS between two half-integer quadrupolar nuclei. AB - Hartmann-Hahn match conditions for n2 --> M2 CP/MAS between two quadrupolar nuclei, spin-lock signal as a function of effective nutation frequency, and the correlation of effective nutation frequency and radiofrequency field strength are reported for three samples: sodium diborate (Na2B4O7), aluminum boride (AlB2), and lithiump aluminate (LiAlO2). Radiofrequency field strengths used for CP/MAS are both greater and less than the sample spinning speed of 10 kHz, resulting in the observation of both zero-quantum and double-quantum matches, which have signals of opposite sign. The match conditions for Na2B4O7 are as expected from published theory and CP/MAS experiments on spins 12 and n2 (n = 3 or 5) with quadrupole frequencies (omegaQ) large compared to the radiofrequency field strength of the CP contact pulse, consisting mainly of sideband matches at one and two times the sample spinning frequency, and the correlation of effective nutation frequency and radiofrequency field strength supports the conclusion that omegaQ is large for both 11B and 23Na. Aluminum-27 in AlB2 may have either small or intermediate omegaQ, and 7Li in LiAlO2 is proposed to have intermediate omegaQ in relation to the radiofrequency field strength, and both have curves of the spin-lock signal as a function of effective nutation frequency with central minima, differing from those of the nuclei with large omegaQ. The sign of the CP/MAS signal for AlB2 and LiAlO2 appears to vary with the CP field strengths for the two nuclei so that positive or negative signals cannot be consistently correlated with zero- or double-quantum matches. However, it is possible to assign at least some of the matches as close to integral multiples of the sample spinning frequency, and some of these are matches at greater than two times the sample spinning frequency. PMID- 10388590 TI - Electron spin transition solution applicable to an ensemble of isolated electrons. AB - An electron spin aligned with a static magnetic field changes its orientation when subjected to a time-varying magnetic field which is directed perpendicular to the static magnetic field. This well-known phenomenon is readily calculated when the time-varying magnetic field is circularly polarized; however, the evolution of the spin-state wavefunctions becomes much more difficult to calculate when the time-varying magnetic field is linearly polarized. For linear polarization and isolated spins, an analytic solution has been derived for the dynamical spin-state wavefunctions. Part of the solution procedure relies on an expansion using a small parameter, which is the ratio of the amplitude of the time-varying magnetic field to the static magnetic field. To verify the validity of the expansion technique, a numerical solution of the basic equations is compared to the analytic solution. Results are found to agree to better than 10% for exact resonance and better than 5% in general. PMID- 10388591 TI - The spin coherence relaxation in the rotating frame as a microscopy parameter for strongly coupled spin systems. AB - The transverse relaxation time in the rotating frame T2rho is proposed as an effective parameter to get specific contrast in solid state imaging. Several peculiarities make T2rho an interesting candidate to map dynamics and structure in solids: the effect of the secular spin interaction can be controlled by the experimenter and therefore the relaxation associated with the nonsecular terms, which is particularly sensitive to very slow dynamics, can be observed. In this paper we present preliminary results obtained on polymers and prove the capability of the MARF Imaging, enhanced by a filter based on rotary echo refocusing, to produce images of solids contrasted by T2rho. PMID- 10388592 TI - Calculation of MAS spectra influenced by slow molecular tumbling. AB - A numeric algorithm is proposed that is suitable to calculate spectral lineshapes influenced by isotropic and anisotropic tumbling under sample spinning conditions. It is based on the stochastic Liouville equation and a rotational diffusion process described by a stationary Markov operator. A corresponding FORTRAN program can be implemented on a regular personal computer. The calculations result in spectral lineshapes including a complete set of spinning sidebands. The sensitive time scale of the resulting lineshapes depends on the deviation of the sample spinning axis from the magic angle. An example is presented demonstrating the potential of off-magic-angle spinning as a tool to analyze slow tumbling motions. PMID- 10388593 TI - Optimization of offset frequency in the SORC pulse sequence using feedback. AB - The low signal-to-noise ratio (SNR) of nuclear quadrupolar resonance measurements has motivated research on signal enhancement methods, including multipulse sequences that facilitate signal averaging, the development of interlaced pulse sequences, and super-Q coils. More recently, it has been shown that feedback can be used to automatically optimize pulse sequence parameters, maximizing the SNR. This paper extends this work by using feedback to optimize the offset frequency in the strong off-resonant comb pulse sequence. Analysis and results are presented for a sample of sodium nitrite at both liquid nitrogen and room temperatures. PMID- 10388594 TI - Berry's phase in the presence of a non-adiabatic environment with an application to magnetic resonance. AB - We consider a two-level system coupled to an environment that evolves non adiabatically. We present a non-perturbative method for determining the persistence amplitude whose phase contains all the corrections to Berry's phase produced by the non-adiabatic motion of the environment. Specifically, it includes the effect of transitions between the two energy levels to all orders in the non-adiabatic coupling. The problem of determining all non-adiabatic corrections is reduced to solving an ordinary differential equation to which numerical methods should provide solutions in a variety of situations. We apply our method to a particular example that can be realized as a magnetic resonance experiment, thus raising the possibility of testing our results in the laboratory. PMID- 10388595 TI - Electron spin lattice relaxation rates for S = 12 molecular species in glassy matrices or magnetically dilute solids at temperatures between 10 and 300 K. AB - The temperature dependence of X-band electron spin-lattice relaxation between about 10 and 300 K in magnetically dilute solids and up to the softening temperature in glassy solvents was analyzed for three organic radicals and 14 S = 12 transition metal complexes. Contributions from the direct, Raman, local vibrational mode, thermally activated, and Orbach processes were considered. For most samples it was necessary to include more than one process to fit the experimental data. Debye temperatures were between 50 and 135 K. For small molecules the Debye temperature required to fit the relaxation data was higher in 1:1 water:glycerol than in organic solvents. For larger molecules the Debye temperature was less dependent upon solvent and more dependent upon the characteristics of the molecule. The coefficients of the Raman process increased with increasing g anisotropy and decreasing rigidity of the molecule. For the transition metal complexes the data are consistent with major contributions from local modes with energies in the range of 185 to 350 K (130 to 240 cm-1). The coefficient for this contribution increases in the order 3d < 4d transition metal. For C-60 anions there is a major contribution from a thermally activated process with an activation energy of about 240 cm-1. For low-spin hemes the dominant contribution at higher temperatures is from a local mode or thermally activated process with a characteristic energy of about 175 cm-1. PMID- 10388596 TI - Novel methods for characterizing a decoupler channel using "undetectable" quantum coherences. AB - Exact solutions for the effect of time-independent RF pulses on any initial configuration of an IS J-coupled system demonstrate that on-resonance CW decoupling yields signals whose frequency depends on RF field strength and homogeneity. These signals are enhanced starting with "undetectable" antiphase and multiple quantum coherences, which can also produce centerband intensity to mimic the signal from decoupled Sx. Conversely, these coherences can be generated from Sx using a low-power pulse, B1 = J/2, of length (2J)-1, dubbed a "90J pulse" since it is the selective equivalent of {(2J)-1-90[I]}. Utilizing 90J pulses, new characterization-of-decoupler (COD) pulse sequences can determine the performance of an insensitive I-spin channel by observing large signals from either antiphase or multiple quantum coherences with the S-spin channel, allowing, in minutes rather than hours: (i) frequency calibration to an accuracy of 0.1 Hz; (ii) measurement of RF amplitudes over a 500-fold variation; and (iii) mapping of RF homogeneity along the sample axis with a single 1D B1 spectrum. These 90J coherence transfer pulses are of potential general use for selective spectroscopy. PMID- 10388597 TI - Determination of 3J(H3i, Pi+1) and 3J(H5i/5i, Pi) coupling constants in 13C labeled nucleic acids using constant-time HMQC. AB - A novel 1H-13C correlated two-dimensional experiment, CT-HMQC-J, for the measurement of three-bond proton-phosphorus coupling constants in 13C-labeled DNA is described. The experiment is based on the intensity difference of 1H-13C cross peaks in the presence and absence of the proton-phosphorus coupling interaction during the constant-time period in HMQC experiment. The 3J(H, P) coupling constants can be easily extracted from the intensity ratios of the two experiments. The method has been applied to a uniformly 13C, 15N-labeled d(GGAGGAT) 7-mer DNA sample. The proton-phosphorus coupling constants determined from CT-HMQC-J, together with the other three-bond coupling constants, are used to determine beta and epsilon torsion angles. The introduction of beta and epsilon restraints has improved the convergence as well as the quality of d(GGAGGAT) structure. PMID- 10388598 TI - A comparison of two controlled-release delivery systems for the delivery of amiloride to control angiogenesis. AB - The diuretic amiloride has been reported to inhibit both Na+-H+ antiport and the urokinase-type plasminogen activator. As a consequence of these inhibitions, neovascularization may also be inhibited. We hypothesized that if amiloride could be effectively delivered in a site-specific manner, a system might be developed that could inhibit localized angiogenesis. In order to evaluate this possibility we conducted a study that compared two different controlled-release systems into which amiloride had been incorporated. The effectiveness of amiloride release from each delivery system was determined by quantitating angiogenic patterns in a chick chorioallantoic membrane (CAM) system using a fractal analysis software program. The two delivery systems compared were sucrose acetate isobutyrate (SAIB) and calcium alginate. Initial HPLC laboratory tests confirmed that amiloride could be released from both SAIB and calcium alginate in vitro in a sustained manner for 72 h. The CAM studies confirmed that neither SAIB nor calcium alginate alone promoted or inhibited angiogenesis when compared to nontreated controls. The release of amiloride from each delivery vehicle resulted in a significant (P < 0.05) inhibition of angiogenesis following both 24 and 48 h of release compared to controls. There was no difference in inhibition of angiogenesis, however, when comparing SAIB + amiloride treated CAMs with calcium alginate + amiloride treated CAMs. These data suggest that both SAIB and calcium alginate may be useful delivery vehicles for the localized application of amiloride to control angiogenesis. Such a system could potentially control tumor angiogenesis without systemic effects. PMID- 10388599 TI - Ultrastructural characteristics of myocardial and coronary microvascular lesions in Kawasaki disease. AB - Kawasaki disease (KD) is an acute febrile mucocutaneous lymph node syndrome with multisystemic vasculitis affecting mainly infants and children. Although several studies on cardiovascular lesions in KD have been done at the light microscopic level, the ultrastructural characteristics and incidences of these lesions have not been well defined. In order to investigate the myocardial and coronary microvascular sequelae in KD, we performed an ultrastructural study on endomyocardial biopsy specimens obtained during follow-up from 54 patients who had typical clinical manifestations of KD, of whom 47 had associated coronary aneurysms as demonstrated by coronary arteriography (CAG) or two-dimensional echocardiography in the acute or healed stage. The average age of onset was 2.2 years old and the duration of illness was from 2 months to 23 years. Follow-up CAG showed that the coronary aneurysms persisted in 33 of the 47 patients (8 with associated stenosis) and resolved in the remaining 14 patients. Ultrastructurally, the myocardial changes revealed hypertrophy, various degrees of degeneration, proliferation and abnormality of mitochondria, infiltration of a small number of lymphocytes, and fibrosis. The coronary microvascular lesion was characterized by microvascular dilatation, endothelial cell injury, platelet aggregation with thrombosis, and stenotic lumen with thickened walls in the small arterioles. It persisted after convalescent stage even up to 23 years and closely correlated with the myocardial sequelae. Moreover, significantly increased incidences of myocardial and coronary microvascular lesions were found in patients with coronary artery lesion. These findings suggest the coronary microvascular lesion as a possible underlying factor of persistent sequelae in KD. PMID- 10388600 TI - The effect of alpha-lipoic acid on the neurovascular reflex arc in patients with diabetic neuropathy assessed by capillary microscopy. AB - Patients with diabetic polyneuropathy are known to have an impaired neurovascular reflex arc compared to healthy controls. This is seen in a delayed decrease in microcirculation of the ipsilateral hand after cooling of the contralateral hand. The aim of this pilot study was to investigate whether intravenous alpha-lipoic acid (ALA) (Thioctacid, Asta Medica) therapy might be able to improve this impaired neurovascular reflex arc in patients with diabetic neuropathy. In addition, clinical effects were evaluated with the aid of the neuropathy symptom score (NSS) and the neuropathy disability score (NDS). Ten patients with diabetes mellitus and polyneuropathy (5 females, 5 males, 2 smokers, 5 IDDM, 5 NIDDM, body mass index 26.1 +/- 1.0 kg/m2, age 58.3 +/- 9.5 years, diabetes duration 15.7 +/- 11.2 years, Hb A1c 6.8 +/- 0.3%) were investigated by nail-fold capillaroscopy after contralateral cooling before and after intravenous therapy with 600 mg alpha-lipoic acid per day over 3 weeks. Cardiac autonomic neuropathy was excluded by beat-to-beat variation analysis. Symptoms of diabetic neuropathy were evaluated before and after therapy with the aid of the NSS and NDS. Capillary blood cell velocity (CBV) of the hand was determined before, during, and for the following 30 min after cooling (3 min at 15 degrees C) of the contralateral hand. Blood pressure, heart rate, and local skin temperature were monitored at 2-min intervals. ALA therapy resulted in a significant improvement of the microcirculatory response to cooling, as seen by an immediate decrease in CBV of 12. 3% (P < 0.02 vs before treatment), which was absent before therapy. Blood pressure, heart rate, and local skin temperature were not different between investigations. There was a significant improvement of the NSS after therapy (5.4 +/- 1.1 vs 8.6 +/- 1.1 points, P < 0.01). These results demonstrate that intravenous therapy with ALA has a positive influence on the impaired neurovascular reflex arc in patients with diabetic neuropathy. PMID- 10388601 TI - Effects of cyclic GMP on microvascular permeability of the cerebral cortex. AB - This study was performed to test the hypothesis that a direct application of cyclic guanosine monophosphate (cGMP) to the cortex would increase blood-brain barrier (BBB) permeability. Rats were anesthetized with 1.4% isoflurane and were mechanically ventilated. Two cranial windows (3 mm in diameter) were made on each side of the rat's skull (a total of four windows on each rat) to expose the cerebral cortex. A patch of normal saline, 10(-5) M, 10(-4) M, or 10(-3) M 8 bromo-cGMP was applied to each cranial window. The patches were changed every 5 min. Ten minutes after applying the patches, BBB permeability was determined by measuring the transfer coefficient (Ki) of [alpha-14C]aminoisobutyric acid. Vital signs were not changed after applying 8-bromo-cGMP. Blood gases were within normal limits. In the cortex, 10(-5) M 8-bromo-cGMP did not significantly affect the Ki; 10(-4) M 8-bromo-cGMP increased the Ki by 115%; 10(-3) M 8-bromo-cGMP increased the Ki by 124%. However, there was no statistical difference in the Ki between the doses of 10(-4) M and 10(-3) M 8-bromo-cGMP. In the pons where no patch was applied, the Ki was similar to that of the cortical area where a normal saline patch was applied. Our data demonstrated that a direct application of cGMP to the cerebral cortex significantly increased the permeability of the BBB. PMID- 10388602 TI - Nonspecific microvascular vasodilation during iontophoresis is attenuated by application of hyperosmolar saline. AB - Iontophoretic administration of acetylcholine chloride (ACh) and sodium nitroprusside (SNP) combined with laser Doppler skin blood perfusion measurements are used for determination of endothelial-dependent and -independent vasodilation. However, the method is biased by nonspecific vasodilation. The primary aim of this study was to investigate if iontophoresis-induced nonspecific vasodilation may be attenuated by addition of high molar concentrations of NaCl to the iontophoresis solutions. Secondary we investigated the applicability of 5 mol/liter NaCl solution as vehicle for ACh and SNP in this method. Skin perfusion changes were determined for iontophoresis of pure vehicles, deionized water and 5 mol/liter NaCl solution, in 12 healthy volunteers. Responses in skin perfusion to iontophoresis of ACh and SNP dissolved in both vehicles were also investigated. Addition of 5 mol/liter NaCl to deionized water significantly attenuated the nonspecific vasodilation and lowered the potential applied over the skin. The inter- and intraindividual coefficients of variation to ACh and SNP responses became, however, higher using hyperosmolar vehicle. During iontophoresis of SNP (in deionized water) we were unable to distinguish between SNP and vehicle effects. This study shows that the nonspecific vasodilation induced by iontophoresis can be attenuated by addition of 5 mol/liter NaCl, possibly due to lower electrical potential over the skin. However, the variability of the method was not improved. When deionized water was used as vehicle the effect of SNP could not be differentiated from that of the vehicle. This was not the case for ACh. PMID- 10388603 TI - Immunohistochemical and ultrastructural characterization of cortical plate microvasculature in the human fetus telencephalon. AB - The blood-brain barrier (BBB) differentiation was investigated by immunohistochemistry and electron microscopy in the radial microvasculature of the telencephalon cortical plate (CP) of 12- and 18-week human fetuses. The BBB specific glucose transporter isoform 1 (GLUT1) is expressed in both stages, with a main localization on the ablumenal and lateral plasma membranes of the endothelial cells. The endothelial cells are welded by short junctions with fusion points of the plasma membranes at 12 weeks and by extensive tight junctions at 18 weeks. The basal lamina is discontinuous beneath the endothelium pericyte layer at 12 weeks and splits into two continuous layers circumscribing the pericytes in the later stage. The expression of laminin, a basal lamina glycoprotein, is continuous already at 12 weeks. The CP microvessels are tightly surrounded by processes of glial cells. Immunodetection of the cytoskeletal filament proteins, vimentin (VIM), and glial fibrillary acidic protein (GFAP), demonstrates that at 12 weeks the perivascular glial processes are mostly represented by VIM-stained fibers of the radial glia. At 18 weeks, GFAP-stained radial glia fibers, processes of VIM-stained astroblasts, and GFAP-positive astrocytes also build the perivascular envelopes. The results indicate that the vessel differentiation is already under way in the human CP at the midgestational age and entails the establishment of some barrier devices. The early relationship between perivascular glia coverage formation and endothelial barrier maturation suggests that also immature astroglial cells are involved in the setting up of the BBB. PMID- 10388604 TI - Red blood cell velocity and volumetric flow assessment by enhanced high resolution laser Doppler imaging in separate vessels of the hamster cheek pouch microcirculation. AB - An enhanced high-resolution laser Doppler imager (EHR-LDI), configured to fit the demands of a measurement area containing separate microvessels, was evaluated for perfusion measurements in hamster cheek pouch preparations during ischemia, reperfusion, and pharmacologically induced vasodilation and vasoconstriction. Measurements in separate microvessels where the laser beam was smaller than the vessel diameter were referred to as red blood cell (RBC) velocity estimates, as previously validated in vitro, whereas a relative flow index, RFI (mean RBC velocity/tissue area), was introduced as a volumetric flow measure. Microvessel diameter and RBC velocity changes during ischemia, reperfusion, as well as during vasoconstriction and vasodilation correlated to the data obtained from the microscope. Correspondingly, during the described provocations anticipated volumetric flow changes were registered as changes in the RFI. When data on intravessel RBC velocity profiles are presented they reflect a parabolic flow profile usually seen in this size microvessel. The EHR-LDI appears a promising tool for investigation of the microvasculature, as it almost simultaneously provides information on relative changes of both in vivo RBC velocity and volumetric flow (RFI), although the latter estimate needs to be further refined. PMID- 10388605 TI - Microvascular permeability change induced by platelet-activating factor is impaired in diabetic mice. PMID- 10388606 TI - Visualization of blood vessels without prior perfusion. PMID- 10388607 TI - Computer Technology Training in the Workplace: A Longitudinal Investigation of the Effect of Mood. AB - How does a person's mood during technology training influence motivation, intentions, and, ultimately, usage of the new technology? Do these mood effects dissipate or are they sustainable over time? A repeated-measures field study (n = 316) investigated the effect of mood on employee motivation and intentions toward using a specific computer technology at two points in time: immediately after training and 6 weeks after training. Actual usage behavior was assessed for 12 weeks after training. Each individual was assigned to one of three mood treatments: positive, negative, or control. Results indicated that there were only short-term boosts in intrinsic motivation and intention to use the technology among individuals in the positive mood intervention. However, a long term lowering of intrinsic motivation and intention was observed among those in the negative mood condition. Copyright 1999 Academic Press. PMID- 10388608 TI - The Role of Nonperformance Factors on Job-Related Relocation Opportunities: A Field Study and Laboratory Experiment. AB - Two studies examined the role of employee gender, marital type (single-earner, dual-earner), and parental status in understanding differential access to job opportunities requiring relocation, as well as possible perceptual processes underlying these effects. A large-scale field study (Study 1) found that married women and employees in dual-earner marriages were provided fewer relocation offers than married men and those in single-earner marriages. A laboratory experiment (Study 2) extended Study 1 by examining the perceptual process by which these nonperformance factors affected relocation opportunities. Again, married women and employees in dual-earner marriages received lower recommendation ratings for jobs requiring relocation compared to married men and single-earners, respectively. Further, decision-makers' perceptions of an applicant's willingness to relocate, family resistance to moving, and ease of adjustment to a geographic move partially mediated the relationship between these nonperformance factors and relocation opportunities. Implications for research and applied practice are discussed. Copyright 1999 Academic Press. PMID- 10388609 TI - All Negative Moods Are Not Equal: Motivational Influences of Anxiety and Sadness on Decision Making. AB - Affective states of the same valence may have distinct, yet predictable, influences on decision processes. Results from three experiments show that, in gambling decisions, as well as in job-selection decisions, sad individuals are biased in favor of high-risk/high-reward options, whereas anxious individuals are biased in favor of low-risk/low-reward options. We argue that these biases occur because anxiety and sadness convey distinct types of information to the decision maker and prime different goals. While anxiety primes an implicit goal of uncertainty reduction, sadness primes an implicit goal of reward replacement. We offer that these motivational influences operate through an active process of feeling monitoring, whereby anxious or sad individuals think about the options and ask themselves, "What would I feel better about ellipsis?" Copyright 1999 Academic Press. PMID- 10388610 TI - Dose levels of 0.01-0.2 microg/kg/day diethylstilbestrol are not suitable for use as a positive control in endocrine toxicity studies. PMID- 10388611 TI - Evaluation of veterinary drug residues in food for their potential to affect human intestinal microflora. AB - The use of veterinary drugs in food-producing animals may result in trace quantities of the drugs or their metabolites being present as residues in food. The effects of veterinary drugs intended for use in food-producing animals on intestinal microflora are evaluated in drug registration since these residues may pose a risk due to their antibiotic activity. This article reviews the different in vivo and in vitro experimental test systems and approaches used by animal health industries, contract laboratories, and regulatory authorities to assess the safety of veterinary drug residues in foods for human consumption. Furthermore, we propose a systematic approach to assess the effects and safety of veterinary drug residues on the human intestinal microflora. PMID- 10388612 TI - The use of benchmark dose methodology with acute inhalation lethality data. AB - Benchmark dose methodology has been proposed as a refinement to the no observed adverse effect level (NOAEL) methods currently used for health risk assessments. We compared log-normal probit and quantal Weibull benchmark concentration (BMC) estimates using 1, 5, and 10% response incidences with inhalation toxicity NOAELs and LOAELs from 120 acute lethality data sets. These studies yielded relatively steep dose-response slopes, which in turn influenced the suitability of selecting response incidences. The mean magnitude of difference between the 95% lower confidence limits (LCLs) for 1, 5, or 10% BMCs and corresponding NOAELs was less than twofold using the probit model and less than fourfold using the Weibull model. BMC estimates at the 10% response exceeded the observed LOAEL in some cases. Maximum likelihood estimates for doses with 1, 5, or 10% responses frequently exceeded LOAELs. The probit model repeatedly gave a better fit for the data compared with the Weibull model, resulting in improved goodness of fit tests and reduced 95% confidence intervals. The 95% LCL appears to be necessary at the 1, 5, or 10% response levels in order to safely estimate a concentration below that resulting in a LOAEL. PMID- 10388613 TI - Exposure endpoint selection in acute dietary risk assessment. AB - Current USEPA Office of Pesticide Program approaches to acute dietary risk assessment do not adequately address uncertainty in the distributional analysis of exposure. This is especially true with respect to regulatory decision points (bright lines) located at the far extreme of the cumulative output distribution. Use of the 99.9th centile as a risk assessment endpoint necessitates confidence in food consumption and residue input distributions that cannot be demonstrated with currently available data and analysis approaches. Even for a pesticide with a rich residue database, data limitations are sufficient to skew results to significantly overestimate exposure. This is compounded when extremes in food consumption are used that go beyond the stated error bounds for the database used. Risk management decision making need not consider endpoints at extremes of exposure output distributions in order for mitigation to be protective of sensitive populations. In fact, such decision making is better informed by utilizing more statistically reliable endpoint selection in the risk assessment process. The richest data content in cumulative exposure distributions occurs in regions well removed from output tails, where the pattern of exposure distribution is driven by the effects of residue concentrations. In contrast, the extreme upper tail of the exposure distribution is data poor and is characterized by high uncertainty reflecting extremes in food consumption patterns. At present, risk managers are better served with exposure endpoints removed from the highly uncertain tails of exposure distributions such as the 99.9th centile bright line. The selection of more appropriate risk management decision points should consider the nature of the distribution, the severity of the effect being assessed, and robustness of the data available for assessing acute dietary risk. PMID- 10388614 TI - Estimating provisional acceptable residues for extralabel drug use in livestock. AB - In 1996, the United States Congress passed legislation (Animal Medicinal Drug Use Clarification Act, AMDUCA), which allows some veterinary or human drugs to be used off label in food-producing animals. In order to implement this Act and protect the U.S. consumer, tolerances or safe concentrations are required before a withdrawal time can be estimated for extralabel drug use. Use of foreign MRLs to satisfy these data needs may not be applicable because of differences in safety standards between the U.S. and other countries. This paper presents strategies that can be used to derive equivalent safe concentrations, referred to as provisional acceptable residues (PARs), that may then be used to estimate drug withdrawal times. Health-based methods are proposed for calculating a PAR for a tissue. Procedure A partitions 50% of the acceptable daily intake (ADI) to edible tissues and reserves the remainder for milk. Procedure B equally partitions the ADI into all edible tissues. Procedure C partitions 50% of the ADI to milk and equally partitions the remaining 50% ADI into edible tissues. Simulations were performed for florfenicol, tetracycline, dexamethasone, azaperone, ivermectin, eprinomectin, and doramectin. In general, these simulations resulted in derivation of conservative PARs, which did not result in daily intakes of residues greater than the health-based ADI. These simulations demonstrated that provided the safe concentrations or equivalent PARs are based on rigorous toxicology safety data (e.g., NOELs, ADIs), the safety of food animal products will not be compromised. It is proposed that these PARs can be used for estimating withdrawal times after extralabel drug use or inadvertent exposure to an environmental contaminant where no approved withdrawal time exists. Finally, implementing similar transparent methods could have a positive impact on international harmonization and trade. PMID- 10388615 TI - Are societal judgments being incorporated into the uncertainty factors used in toxicological risk assessment? AB - The aim of this paper is to show that the uncertainty factors used in toxic risk assessment to develop exposure standards do contain societal judgments as well as technical judgments. The process generally used today originated in the 1950s, when a deterministic approach to risk was the norm. Technical judgments are required concerning the nature and the quality of the evidence used in the risk assessment. Judgments taken are essentially cautious. This caution may not matter when measured exposure is significantly below the standard and may be accepted when exposure occurs only following an approval process based on "gate keeping." More sophisticated judgments are required when actual exposure may exceed this type of standard or when risk needs to be compared with benefit. These circumstances can occur with patient exposure to human medicines and with occupational exposure to chemicals. Under these circumstances more explicitly considered societal judgments concerning what constitute "broadly acceptable" and "tolerable" risk criteria, and hence what are appropriate uncertainty factors, are required. The outcomes of those societal judgments are likely to vary according to the circumstances surrounding the exposure and have led to smaller uncertainty factors being considered appropriate for occupational exposure, when compared with widespread public exposure. PMID- 10388616 TI - Evaluation of the EPA exposure assessment for phosphoric acid. AB - The philosophy of this paper is that the Environmental Protection Agency's (EPA) 1990 exposure assessment for phosphoric acid is not based on sound science and therefore does not accurately characterize the eutrophication potential of phosphoric acid releases from Toxic Release Inventory (TRI) reporting facilities. This error propagates the myth that eutrophication is triggered by the release of phosphoric acid from facilities reporting releases in the TRI. A detailed evaluation of EPA's 1990 exposure assessment for phosphoric acid reveals critical flaws in their applied methodology. Significant methodological improvements detailed in this paper fundamentally alter EPA's 1990 exposure assessment and demonstrate that, contrary to EPA's conclusion, these sources of phosphoric acid do not have the potential to trigger eutrophication in surface waters. Therefore, when sound scientific principles are applied in this comparative exposure assessment, EPA's policy maintaining phosphoric acid on the TRI list is inappropriate. PMID- 10388617 TI - Chemical control laws on polymers: A case for harmonization. AB - All industrial countries, and several emerging and developing countries, have established regulations to control chemical substances, with a view toward limiting the risks to workers and the public, as well as reducing the impact on the environment. However, the lack of harmonization among such international schemes is creating international trade difficulties. In this context, polymers, and particularly specialty polymers, are a unique case. Polymers are less easily characterized than other substances, and parameters used by governments to control them may vary greatly, leading to national regulations that are significantly dissimilar, or even incompatible. This paper reviews chemical control laws on polymers, identifies the major differences between them, and suggests model legislation for polymers that would reduce barriers to international trade in polymers while maintaining a high degree of human and environmental protection. PMID- 10388618 TI - A cancer risk assessment of di(2-ethylhexyl)phthalate: application of the new U.S. EPA Risk Assessment Guidelines. AB - The current United States Environmental Protection Agency (EPA) classification of di(2-ethylhexyl)phthalate (DEHP) as a B2 "probable human" carcinogen is based on outdated information. New toxicology data and a considerable amount of new mechanistic evidence were used to reconsider the cancer classification of DEHP under EPA's proposed new cancer risk assessment guidelines. The total weight-of evidence clearly indicates that DEHP is not genotoxic. In vivo administration of DEHP to rats and mice results in peroxisome proliferation in the liver, and there is strong evidence and scientific consensus that, in rodents, peroxisome proliferation is directly associated with the onset of liver cancer. Peroxisome proliferation is a transcription-mediated process that involves activation by the peroxisome proliferator of a nuclear receptor in rodent liver called the peroxisome proliferator-activated receptor (PPARalpha). The critical role of PPARalpha in peroxisomal proliferation and carcinogenicity in mice is clearly established by the lack of either response in mice genetically modified to remove the PPARalpha. Several mechanisms have been proposed to explain how, in rodents, peroxisome proliferation can lead to the formation of hepatocellular tumors. The general consensus of scientific opinion is that PPARalpha-induced mitogenesis and cell proliferation are probably the major mechanisms responsible for peroxisome proliferator-induced hepatocarcinogenesis in rodents. Oxidative stress appears to play a significant role in this increased cell proliferation. It triggers the release of TNFalpha by Kupffer cells, which in turn acts as a potent mitogen in hepatocytes. Rats and mice are uniquely responsive to the morphological, biochemical, and chronic carcinogenic effects of peroxisome proliferators, while guinea pigs, dogs, nonhuman primates, and humans are essentially nonresponsive or refractory; Syrian hamsters exhibit intermediate responsiveness. These differences are explained, in part, by marked interspecies variations in the expression of PPARalpha, with levels of expression in humans being only 1-10% of the levels found in rat and mouse liver. Recent studies of DEHP clearly indicate a nonlinear dose-response curve that strongly suggests the existence of a dose threshold below which tumors in rodents are not induced. Thus, the hepatocarcinogenic effects of DEHP in rodents result directly from the receptor mediated, threshold-based mechanism of peroxisome proliferation, a well understood process associated uniquely with rodents. Since humans are quite refractory to peroxisomal proliferation, even following exposure to potent proliferators such as hypolipidemic drugs, it is concluded that the hepatocarcinogenic response of rodents to DEHP is not relevant to human cancer risk at any anticipated exposure level. DEHP should be classified an unlikely human carcinogen with a margin of exposure (MOE) approach to risk assessment. The most appropriate and conservative point of reference for assessing MOEs should be 20 mg/kg/day, which is the mouse NOEL for peroxisome proliferation and increased liver weight. Exposure of the general human population to DEHP is approximately 30 microg/kg body wt/day, the major source being from residues in food. Higher exposures occur occupationally [up to about 700 microg/kg body wt/day (mainly by inhalation) based on current workplace standards] and through use of certain medical devices [e.g., up to 457 microg/kg body wt/day for hemodialysis patients (intravenous)], although these have little relevance because the routes of exposure bypass critical activation enzymes in the gastrointestinal tract. PMID- 10388619 TI - Crystallization notes PMID- 10388620 TI - Crystal structure of a phycourobilin-containing phycoerythrin at 1.90-A resolution. AB - The structure of R-phycoerythrin (R-PE) from the red alga Griffithsia monilis was solved at 1.90-A resolution by molecular replacement, using the atomic coordinates of cyanobacterial phycocyanin from Fremyella diplosiphon as a model. The crystallographic R factor for the final model is 17.5% (Rfree 22.7%) for reflections in the range 100-1.90 A. The model consists of an (alphabeta)2 dimer with an internal noncrystallographic dyad and a fragment of the gamma polypeptide. The alpha-polypeptide (164 amino acid residues) has two covalently bound phycoerythrobilins at positions alpha82 and alpha139. The beta-polypeptide (177 residues) has two phycoerythrobilins bound to residues beta82 and beta158 and one phycourobilin covalently attached to rings A and D at residues beta50 and beta61, respectively. The electron density of the gamma-polypeptide is mostly averaged out by threefold crystallographic symmetry, but a dipeptide (Gly-Tyr) and one single Tyr could be modeled. These two tyrosine residues of the gamma polypeptide are in close proximity to the phycoerythrobilins at position beta82 of two symmetry-related beta-polypeptides and are related by the same noncrystallographic dyad as the (alphabeta)2 dimer. Possible energy transfer pathways are discussed briefly. PMID- 10388621 TI - Isolation, electron microscopic imaging, and 3-D visualization of native cardiac thin myofilaments. AB - An increasing number of cardiac diseases are currently pinpointed to reside at the level of the thin myofilaments (e.g., cardiomyopathies, reperfusion injury). Hence the aim of our study was to develop a new method for the isolation of mammalian thin myofilaments suitable for subsequent high-resolution electron microscopic imaging. Native cardiac thin myofilaments were extracted from glycerinated porcine myocardial tissue in the presence of protease inhibitors. Separation of thick and thin myofilaments was achieved by addition of ATP and several centrifugation steps. Negative staining and subsequent conventional and scanning transmission electron microscopy (STEM) of thin myofilaments permitted visualization of molecular details; unlike conventional preparations of thin myofilaments, our method reveals the F-actin moiety and allows direct recognition of thin myofilament-associated porcine cardiac troponin complexes. They appear as "bulges" at regular intervals of approximately 36 nm along the actin filaments. Protein analysis using SDS-polyacrylamide gel electrophoresis revealed that only approximately 20% troponin I was lost during the isolation procedure. In a further step, 3-D helical reconstructions were calculated using STEM dark-field images. These 3-D reconstructions will allow further characterization of molecular details, and they will be useful for directly visualizing molecular alterations related to diseased cardiac thin myofilaments (e.g., reperfusion injury, alterations of Ca2+-mediated tropomyosin switch). PMID- 10388622 TI - Surface ultrastructure of pit organ, spectacle, and non pit organ epidermis of infrared imaging boid snakes: A scanning probe and scanning electron microscopy study. AB - Boid snakes possess unique infrared imaging pit organs. The ultrastructure of the surfaces of these organs scatter or reflect electromagnetic radiation of specific wavelengths. Pit organ epidermal surfaces of boid snakes are covered with arrays of pore-like structures called micropits. In order to determine the dimensions of this complicated surface structure, we have performed the first ultrastructural analysis on snake epidermis by high-resolution microscopy techniques. Using scanning probe microscopy and scanning electron microscopy, we found that the epidermis of pit organ, maxillary non pit organ, spectacle, and ventral scales contain arrays of micropits. These scale surfaces also contain major surface features of overlapping plate-like structures. Pit organ micropits averaged 319 nm in diameter and 46 nm in depth and were spaced an average of 808 nm from each other. These micropits were significantly deeper, of greater diameter, and spaced at greater distances apart than those of the other scales. Plate structures of the pit organs had a mean distance between plates of 3.5 microm and a mean plate step height of 151 nm. These differences serve to strengthen the argument that arrays of micropit and plate surface structures function as spectral filters or anti-reflective coatings with respect to incident electromagnetic radiation. PMID- 10388623 TI - Partially systematic molecular packing in the hexagonal columnar phase of dogfish egg case collagen. AB - The collagen that forms the egg case of the dogfish Scyliorhinus canicula is stored in bulk in the female nidamental glands. Here the collagen molecules are thought to undergo a series of distinct pH-dependent liquid crystalline aggregation phase changes before assembling into the final arrangement encountered in the mature egg case. One liquid crystalline phase is hexagonal with the centres of two adjacent hexagons about 36 nm apart. We have collected tilt series of the hexagonal phase from plastic sections of the nidamental gland and have produced a three-dimensional reconstruction of the collagen arrangement of this phase. The reconstruction features axial columns of protein density lying regularly on the vertices of hexagonal cells of edge length 21 nm. Each column is connected to three nearest neighbours by irregular sheets of protein, but there appear to be preferred molecular directions at about 40 degrees to 50 degrees to the columns. The reconstruction has been interpreted in terms of known interactions of this collagen in other assemblies. PMID- 10388624 TI - The crystal structure of the SIV gp41 ectodomain at 1.47 A resolution. AB - Cell membrane fusion by human (HIV) and simian (SIV) immunodeficiency viruses is mediated by the envelope glycoproteins gp120 and gp41. Although the precise mechanism of the fusion process is unknown, the ectodomain of gp41 is thought to undergo dramatic rearrangement from its prefusogenic state. To elucidate this process further, the crystal structure of the SIV gp41 ectodomain (residues 27 149) was determined at 1.47 A resolution and is reported herein. It is the most accurate and complete structure of a retroviral gp41 ectodomain determined to date. The rod-like trimeric structure of SIV gp41 comprises three parallel N terminal alpha-helices assembled as a coiled coil in the center with three antiparallel C-terminal alpha-helices packed on the outside connected by highly flexible loops. Portions of the loops in all three monomers are crystallographically disordered and could not be accurately modeled. The core of the structure is similar (but not identical) to those of smaller HIV/SIV gp41 segments previously determined by X-ray crystallography with root mean square deviations in main chain atoms of less than 1.0 A. The crystal structure differs more substantially from the reported NMR solution structure of the identical SIV construct. The mechanisms of viral fusion and the inhibition by peptides are discussed in the context of the three-dimensional structure. PMID- 10388625 TI - An 8-A projected structure of FhuA, A "ligand-gated" channel of the Escherichia coli outer membrane. AB - The structure of FhuA, a siderophore and phage receptor in the outer membrane of Escherichia coli, has been investigated by electron crystallography. Bidimensional crystals of hexahistidine-tagged FhuA protein solubilized in N,N dimethyldodecylamine-N-oxide were produced after detergent removal with polystyrene beads. Frozen-hydrated crystals (unit cell dimensions of a = 124 A, b = 98 A, gamma = 90 degrees ) exhibited a p22121 plane group symmetry. A projection map at 8 A resolution showed the presence of dimeric ring-like structures with an elliptical shape (48 x 40 A). Each monomer was composed of a ring of densities with a radial width of 8-10 A corresponding to a cylinder of beta sheets. Few densities are present inside the barrel, leaving a central channel approximately 25 A in diameter. A projection map of FhuA at 15 A resolution, which was calculated from negatively stained preparations, demonstrated that most of the central channel was masked by extramembrane domains. This map also revealed an asymmetric distribution of extramembrane domains in FhuA, with large domains located mainly on one side of the molecule. Comparison with density maps derived from recent atomic structure allowed further interpretation of the electron microscopy projection structures with regard to long hydrophilic loops governing the selectivity and opening of the channel. PMID- 10388627 TI - Biochemical and crystallographic characterization of a Rho effector domain of the protein serine/threonine kinase N in a complex with RhoA. AB - The effector domain of human protein serine/threonine kinase N (PKN), an effector protein for the small GTP-binding protein Rho, was expressed and purified for protein characterization and crystallization in a complex form with human RhoA. In solution, RhoA binds to the PKN effector domain with 1:2 stoichiometry in a GTP-dependent manner. The obtained complex crystals diffract to 2.2 A resolution. PMID- 10388626 TI - Structural studies on a 2,3-diphosphoglycerate independent phosphoglycerate mutase from Bacillus stearothermophilus. AB - Phosphoglycerate mutase (PGM), an important enzyme in the glycolytic pathway, catalyzes the transfer of a phosphate group between the 2 and the 3 positions of glyceric acid. The gene coding for the 2, 3-diphosphoglycerate independent monomeric PGM from Bacillus stearothermophilus (57 kDa), whose activity is extremely pH sensitive and has an absolute and specific requirement for Mn2+, has been cloned and the enzyme overexpressed and purified to homogeneity. Circular dichroism studies showed at most only small secondary structure changes in the enzyme upon binding to Mn2+ or its 3-phosphoglycerate substrate, but thermal unfolding analyses revealed that Mn2+ but not 3-phosphoglycerate caused a large increase in the enzyme's stability. Diffraction-quality crystals of the enzyme were obtained at neutral pH in the presence of 3-phosphoglyceric acid with ammonium sulfate as the precipitating agent; these crystals diffract X rays to beyond 2.5-A resolution and belong to the orthorhombic space group C2221 with unit cell dimensions, a = 58.42, b = 206.08, c = 124.87 A, and alpha = beta = gamma = 90.0 degrees. The selenomethionyl version of the B. stearothermophilus protein has also been overexpressed, purified, and crystallized. Employing these crystals, the determination of the three-dimensional structure of this PGM by the multiwavelength anomalous dispersion method is in progress. PMID- 10388628 TI - Crystallization and preliminary X-ray diffraction analysis of a recombinant bacterial heme oxygenase (Hmu O) from Corynebacterium diphtheriae. AB - Hmu O is a 24-kDa soluble bacterial heme degradation enzyme found in the pathogen Corynebacterium diphtheriae, the causative agent of diphtheria. Similar to the mammalian heme oxygenase, it binds hemin stoichiometrically and catalyzes the oxygen-dependent conversion of hemin to biliverdin, carbon monoxide, and free iron. Iron is an essential nutrient for bacteria and especially important for pathogenesis. Here we report the first crystallization and preliminary crystallographic study of the heme-Hmu O complex formed from hemin and a recombinant Hmu O, which was expressed in Escherichia coli from a synthetic gene based on the putative hmu O gene sequence. Crystals of the heme-Hmu O complex were obtained by the sitting drop vapor diffusion method using a precipitant solution containing 18% (w/v) PEG 8000 and 0.2 M calcium acetate in 0.1 M sodium cacodylate (pH 6.5). Using synchrotron radiation, the heme-Hmu O crystal diffracted to 2.8 A resolution. It belongs to the monoclinic space group C2, with unit cell parameters a = 123.18 A, b = 44.51 A, c = 92.10 A, and beta = 123.3 degrees. Assuming one molecule of the heme-Hmu O complex per asymmetric unit, the calculated value of Vm is 2.89 A3/Da. PMID- 10388629 TI - Crystallization and preliminary X-ray studies of the Ia component of Clostridium perfringens iota toxin complexed with NADPH. AB - A recombinant Ia component of Clostridium perfringens iota toxin, which ADP ribosylates actin, was crystallized by the hanging drop vapor diffusion method using PEG4000 as a precipitating agent. The crystals were obtained in the presence of NADPH, which is similar to a real substrate, NADH, and belongs to the space group P1 (a = 47.9 A, b = 54.5 A, c = 103.1 A, alpha = 99.0 degrees, beta = 93.3 degrees, and gamma = 107.2 degrees ). The Matthews coefficient of native crystal was 2.7, assuming 2 mol/asymmetric unit. Native data were collected at 2.4-A resolution. The results from a heavy-atom search showed that lanthanide ions (samarium, holmium) altered the molecular packing, judging from the unit cell difference. The crystals also belonged to the space group P1 (a = 47.7 A, b = 53.9 A, c = 54.6 A, alpha = 68.9 degrees, beta = 78.3 degrees, and gamma = 73.7 degrees ), which is consistent with only one molecule per asymmetric unit. PMID- 10388630 TI - Training--vive la difference? PMID- 10388631 TI - Cochrane collaborative review group on peripheral vascular diseases: review abstracts PMID- 10388632 TI - Carotid endarterectomy after recent cerebral infarction. AB - OBJECTIVES: whether timing of carotid endarterectomy (CEA) was significant in terms of morbidity and mortality for significant carotid stenosis. DESIGN/MATERIALS: comparison was made of patients requiring CEA performed in less than 6 weeks or more than 6 weeks after their stroke. To enable quantification in terms of clinical presentation, aetiology and handicap, standardised scales were incorporated into the registry protocol. A postoperative event was considered to have occurred if a stroke or death from any cause took place within one month of surgery. RESULTS: patients with CEA (n=1005) and stroke numbered 232. Comparison was made of the early (n=86) and late surgery groups (n=121) in terms of demography, risk factors, clinical findings, quantitative neurological deficit, handicap and degree of carotid stenosis with no significant differences found except for race. There was no difference in morbidity and mortality (M+M) between the early and late surgery group. The relative risk (RR) of >6 week group was 1.90 (CI: 0.52-6. 94) with an odds ratio of 1.96 (CI: 0.45-9.63). There is, therefore, a trend of a two-fold risk of MM in the >6 week group. CONCLUSION: we propose that the historical 6-week wait period for CEA post stroke is outdated. PMID- 10388633 TI - Ischaemia and reperfusion during open abdominal aortic aneurysm surgery induce extensive thrombin generation and activity. AB - OBJECTIVES: does open surgery for abdominal aortic aneurysm (AAA) influence coagulation? METHODS: in 23 patients operated on for AAA, cubital blood was sampled pre-, intra- and postoperatively. Femoral blood was also sampled intraoperatively. RESULTS: preoperatively, prothrombin fragment 1+2 (F1+2), thrombin-antithrombin complex (TAT) and soluble fibrin (SF) were elevated in AAA patients. During aortic clamping all parameters increased significantly in cubital blood (p<0.01) as well as in femoral blood (p<0.001) and after aortic declamping F1+2 and TAT increased further. F1+2, TAT and SF were significantly higher in femoral than cubital blood. Postoperatively F1+2 and TAT returned to preoperative values, while SF still had a significantly higher level than preoperatively (p<0.001). Blood loss showed co-variation with F1+2 increase in femoral blood after aortic declamping (p<0.05). CONCLUSIONS: these data indicate that the coagulation system was strongly activated by the occurrence of an AAA. During AAA surgery a further extensive activation was seen. The activity was still high, but on decline, one week postoperatively. Ischaemia and reperfusion of the lower part of the body were the major stimuli for thrombin generation and activity. PMID- 10388634 TI - Identification and selective perfusion of the spinal cord-feeding arteries by intrathecal pO2 monitoring for spinal cord protection. AB - OBJECTIVES: to study whether spinal cord-feeding arteries could be identified by the changes in the intrathecal pO2 (I-pO2), and to examine whether selective perfusion of feeding arteries identified by this method could protect the spinal cord against ischaemia. DESIGN: controlled animal experiments. MATERIALS AND METHODS: in experiment 1, using 16 mongrel dogs, 18 segmental arteries were cannulated through which oxygenated saline was injected and the I-pO2 change was observed. When the I-pO2 increase was more than 0.5 mmHg, the artery was considered to be a spinal cord-feeding artery. In experiment 2, involving 10 dogs, the segmental arteries identified as spinal cord-feeding arteries were perfused with arterial blood and the recovery of I-pO2 and evoked spinal potentials (ESP) was examined. RESULTS: of 208 segmental arteries examined, 176 (84.6%) arteries were correctly judged and 32 (15.4%) were not. It was observed that the I-pO2 recovered from 13.9 to 30.5 mmHg and the ESP recovered from 20.9% and 8.2% to 66.5% and 44.7% of each control for the first negative (N1) and second negative (N2) components, respectively. CONCLUSION: spinal cord-feeding arteries were successfully identified using the I-pO2 monitoring method. Perfusion of these arteries with arterial blood improved the I-pO2 and ESP, which were significantly depressed by ischaemia. PMID- 10388635 TI - The influence of experience on the reproducibility of the ankle-brachial systolic pressure ratio in peripheral arterial occlusive disease. AB - OBJECTIVES: to estimate the intra-observer variability of the measurement of the ankle-brachial systolic pressure index (ABPI) and to compare the reproducibility of the measurements by experienced vascular laboratory assistants and by less experienced general practice personnel. DESIGN: repeated measurement of ABPI by general practitioners (GPs), GP-assistants and vascular laboratory assistants using a pocket Doppler device and a random-zero sphygmomanometer. METHODS AND MATERIALS: ABPI was measured in six patients with various degrees of PAOD by two experienced observers (vascular laboratory assistants) and by 24 less-experienced observers (18 practice assistants, six GPs). RESULTS: the total number of measurements was 354. The overall intra-observer variability estimate was 11.8% ABPI. The intra-observer variability was 7.3% in the experienced observers and 12.0% in the less-experienced observers. The difference of variability between experienced and less-experienced observers was significant. CONCLUSIONS: the ABPI is suitable in follow-up studies where repeated measurements are needed. Differences between measurements can be minimised by performing repeated measurements or by using more experienced observers. PMID- 10388636 TI - The morbidity of secondary vascular access. A lifetime of intervention. AB - OBJECTIVES: to examine the outcome and incidence of access-related procedures in patients who dialyse via PTFE secondary access grafts. DESIGN: retrospective case note study. RESULTS: recipients of secondary access procedures face a lifetime of haemodialysis access interventions. In total, 639 access-related procedures were performed on the 72 patients studied. At the end of the study five patients were wholly reliant on central venous catheters for dialysis access. Patients with secondary access grafts have little hope of transplantation; only six of 72 patients received a transplant after a secondary access procedure. CONCLUSION: the increasing number of patients coming to synthetic access-graft procedures and the morbidity of such procedures mean that surgeons should adopt strategies to minimise the use of grafts and limit the number of interventions performed. Careful planning will also reduce the number of central line placements and help to reduce the morbidity associated with long-term haemodialysis. Increasing resources will be required to meet the rising demand for secondary access provision. PMID- 10388637 TI - The influence of indomethacin on the metabolism and cytokine secretion of human aneurysmal aorta. AB - INTRODUCTION: inflammation and proteolysis are important processes in the development of abdominal aortic aneurysms (AAAs). Prostaglandin E2 (PGE2) (a product of cyclo-oxygenase 2), other inflammatory mediators and proteolytic enzymes are produced in high quantities in the aneurysm wall. We developed an explant culture system for AAA tissue to assess the effects of potential drug therapies. METHODS: full thickness biopsies of human AAA were established in culture in the presence or absence of indomethacin (a cyclo-oxygenase-2 inhibitor). The conditioned medium was collected at 48 h intervals and analysed for products of collagen breakdown, matrix metalloproteinases, PGE2 and inflammatory cytokines. Explant viability was assessed by histology, glucose consumption, lactate dehydrogenase release and demonstration of protein synthesis in the tissue. RESULTS: nuclear morphology was maintained for 4 or more days and this, together with biochemical assays, indicated that AAA explants were viable in short-term culture. Indomethacin (10 microM) markedly reduced AAA explant production of prostaglandin E2 from 320 ng/ml to 3.3 ng/ml (p=0.028, n=6). Indomethacin also reduced the release of interleukin-1beta (IL-1beta) (from 166 pg/ml to 9.8 pg/ml, p =0.04, n=5) and interleukin-6 (IL-6) (from 119 ng/ml to 57 ng/ml, p=0.028, n=6), but had no effect on monocyte chemotactic protein 1 or matrix metalloproteinase-9 secretion. CONCLUSIONS: short-term explants of AAA are a novel method to assess the effects of drugs on aneurysm tissue. Indomethacin reduces the production of PGE2, IL-1beta and IL-6, suggesting that cyclo oxygenase-2 inhibitors may control the inflammation in the aneurysm wall and potentially limit AAA growth. PMID- 10388638 TI - Semi-closed femoropopliteal thromboendarterectomy: a prospective study. AB - OBJECTIVE: to study the primary patency rates of angioscopically controlled thromboendarterectomies of the superficial femoral artery. DESIGN: prospective open study. METHODS: between 1990 and 1995, femoropopliteal thromboendarterectomies were performed in 63 patients (41 male, 22 female). Postoperative follow up was performed at 3- to 6-month intervals using non invasive pressure measurements plus IVDSA at 1 year. RESULTS: eight patients were not evaluable, leaving 55 patients eligible for follow-up analysis. Postoperative complications (arteriovenous fistulas, false aneurysms) were observed in 5.4% of patients. Immediate perioperative occlusions occurred in 7.3%, early occlusions in 21.8% and late occlusions in 16.4% of all cases. The mean follow-up was approximately 57 months. The mean primary patency rate at 5 years was 44.5% (28 patients with the superficial femoral artery still open). Six patients died during the follow-up period. CONCLUSIONS: in contrast to the very positive reports found in recent literature, this prospective study shows a lower five year patency rate for semi-closed femoropopliteal thromboendarterectomy than for bypass grafting. Thromboendarterectomy cannot be considered as a standard procedure in revascularisation of the femoropopliteal region. PMID- 10388639 TI - In vivo microscopic study of microcirculatory perfusion of the skin of the foot in peripheral vascular disease. AB - OBJECTIVES: the aim of this study was to determine the proportion of perfused capillaries in the skin of the foot in patients with peripheral vascular disease, and compare it with that in normal subjects. DESIGN: experimental study comparing capillary perfusion in nine patients with severe peripheral vascular disease (group 2) with seven age- and sex-matched control subjects (group 1). MATERIALS AND METHODS: using in vivo video microscopy, a method was developed to measure the ratio of perfused to total capillaries, by comparing the numbers of corresponding capillaries before and after intravenous injection of sodium fluorescein. RESULTS: the mean percentage ratio of perfused to total capillaries was 54.7% (range 41-87%, standard deviation 16.5) in group 1, and 86.0% (range 62 100%, standard deviation 13.2) in group 2 (p<0.001, t-test). CONCLUSION: a significantly higher proportion of capillaries is perfused in the skin of the foot of patients with severe peripheral vascular disease than in that of normal subjects. This is of important pathophysiological significance and may have clinical implications with regard to the role of pharmacological intervention in severe limb ischaemia. PMID- 10388641 TI - Complications after carotid endarterectomy are related to surgical errors in less than one-fifth of cases. Swedvasc--The Swedish Vascular Registry and The Quality Committee for Carotid Artery Surgery. AB - OBJECTIVES: to study possible relations between indications, contraindications and surgical technique and stroke and/or death within 30 days of carotid endarterectomy (CEA). DESIGN: analysis of hospital records for patients identified in a national vascular registry. METHOD: during 1995-1996, 1518 patients were reported to the Swedish Vascular Registry - Swedvasc. Among these the sixty-five with a stroke and/or death within 30 days were selected for study. Complete surgical records were reviewed by three approved reviewers using predetermined criteria for indications and possible errors. RESULTS: an error of surgical technique or postoperative management was found in eleven patients (17%). In six cases (9%) the indication was inappropriate or there was an obvious contraindication. The indication was questionable in fourteen (21.5%). Half of the patients (52.5%) had surgery for an appropriate indication, and no contraindication or error in surgical technique or management was identified. CONCLUSION: more than half the complications of CEA represent the "method cost", i.e. the indication, risk and surgical technique were correct. However, the stroke and/or death rate might be reduced if all operations conformed to agreed criteria. PMID- 10388640 TI - Intraoperative assessment of technical perfection in carotid endarterectomy: a prospective analysis of 1305 completion procedures. Collaborators of the EVEREST study group. Eversion versus standard carotid endartectomy. AB - OBJECTIVE: to define the incidence of technical defects and the impact of technical errors on ipsilateral carotid occlusion, ipsilateral stroke, and early restenosis rates, we analysed 1305 patients undergoing carotid completion procedures. DESIGN: prospective multicentre study. PATIENTS AND METHODS: adequacy of CEA was assessed intraoperatively by angiography in 1004 (77%), by angioscopy in 299 (22%), and by duplex scan in two patients (1%). Arteriograms and angioscopic findings were interpreted at the time of the procedure by the operating surgeon, who also established the need for immediate surgical revision. RESULTS: perioperatively, 13 major strokes (0.9%, all ipsilateral) and six deaths (0.4%) were recorded. Overall, 112 defects (9%) were identified intraoperatively: 81 (72%) were located in the common carotid artery (CCA) or internal carotid artery (ICA), and 31 (28%) in the external carotid artery. In 48 patients (4%) the defects were revised. Logistic regression analysis revealed that carotid plaque extension >2 cm on the ICA was a positive independent predictor of CEA defects (odds ratio (OR) 1.5p=0.03). A significant association was found between the incidence of revised defects of the CCA and ICA and perioperative ipsilateral stroke (OR 11.5p=0.0002). In contrast, patients with minor non-revised defects had an ipsilateral stroke rate comparable to that of patients with no defects (p=0.4). No significant association was found between revised or non-revised defects and occurrence of restenosis/occlusion at 6-month follow-up. CONCLUSIONS: the incidence of major technical defects during CEA is low, yet the perioperative neurological prognosis of patients with major defects warranting revision is poor. Completion angiography or angioscopy and possible correction of defects did not protect per se from an unfavourable early outcome after CEA. Therefore, surgical excellence is mandatory to achieve satisfactory results after CEA. PMID- 10388642 TI - The role of type III collagen in the development of familial abdominal aortic aneurysms. AB - OBJECTIVES: to evaluate the prevalence of familiar abdominal aortic aneurysm (AAA) and the role of type III collagen deficiency. METHODS: fifty-six consecutive patients coming for aneurysm repair were asked if one or more first degree relatives had an AAA. During operation, a skin biopsy was taken from the patients for protein analysis to measure the type III collagen production in cultured fibroblasts. RESULTS: a positive family history was found in 28.6% of the AAA patients. Six (10.7%) of the AAA patients had a type III collagen deficiency (mean 4.3% (S.D.+/-0.5)). In this group three men, mean age 65.3 years (S.D.+/-5.0), had a positive family history and a type III collagen deficiency. Segregation analysis with an intragenic marker in the type III collagen gene in a single family was in favour of linkage with the gene for type III procollagen (COL3A1) locus. CONCLUSIONS: the high prevalence of familial AAA suggests a genetic aetiology. A small group of patients have a type III collagen deficiency. Linkage with the COL3A1 gene could not be proven or excluded in the families studied. PMID- 10388643 TI - Digital photoplethysmography in the diagnosis of suspected lower limb DVT: is it useful? AB - OBJECTIVE: to determine the role of digital photoplethysmography (D-PPG) in the diagnosis of deep-vein thrombosis (DVT), in comparison to the "gold standard" of either contrast ascending venography (ACV) or colour-flow duplex imaging (CFDI). METHOD: prospective study of 100 hospital inpatients (103 legs) referred to the X ray department for ACV or CFDI with clinically suspected lower limb DVT in a district general hospital. Each patient was assessed by either ACV or CFDI, and D PPG. RESULTS: thirty-seven limbs were found to have DVT as demonstrated by ACV or CFDI. All patients with a venous refilling time (RT) of greater than 20 s and venous pump (VP) of greater than 35 had a normal ACV or CFDI. Using RT of less than 21 s as the optimal cut-off point, D-PPG achieved a sensitivity of 100%, negative-predictive value of 100%, specificity of 47% and positive-predictive value of 51%. By using VP of less than 36 as the optimal cut-off point, a sensitivity of 100%, a negative-predictive value of 100%, a specificity of 35% and positive-predictive value of 46% were achieved. CONCLUSIONS: these results validate the use of portable D-PPG as a useful screening tool for the diagnosis of clinically suspected lower limb DVT. A positive test requires further confirmation by one of the "gold standard" methods, whereas a negative test effectively excludes DVT. PMID- 10388644 TI - Endovascular repair of a traumatic innominate artery aneurysm. PMID- 10388645 TI - Middle aortic syndrome: some diagnostic and therapeutic considerations. PMID- 10388646 TI - Campylobacter fetus infection of a previously excluded popliteal aneurysm. PMID- 10388647 TI - Chronic leg pain - problem solved. PMID- 10388648 TI - Support plasmids and support proteins required for recovery of recombinant respiratory syncytial virus. AB - Respiratory syncytial virus (RSV) can be recovered from plasmids that separately encode antigenomic RNA and the N, P, L, and M2-1 proteins of the nucleocapsid. However, in a recent study the inclusion of a separate M2-1 expression plasmid was found to be unnecessary (H. Jin, D. Clarke, H. Zhou, X. Cheng, K. Coelingh, M. Bryant, and S. Li, Virology 1998, 251, 206-214). This suggested that the M2-1 protein, which is a transcription antitermination factor, is not required to reconstitute the minimum unit of infectivity, namely a nucleocapsid fully functional for viral transcription and RNA replication. Here we show that the antigenomic plasmid is remarkably efficient as a substitute for an M2-1 expression plasmid in supporting processive transcription by an RSV minigenome. Thus, the simple expedient of omitting an expression plasmid is invalid for evaluating recovery requirements. The issue of the requirement of M2-1 for the recovery of infectious RSV is discussed. PMID- 10388649 TI - Augmentation of human influenza A virus-specific cytotoxic T lymphocyte memory by influenza vaccine and adjuvanted carriers (ISCOMS). AB - There is a need to improve the ability of subunit vaccines to induce CD8(+) CTL responses in humans, especially for vaccines used to prevent illness by organisms that undergo antigenic variation at their major neutralizing antibody sites, e.g., influenza A viruses and human immunodeficiency virus. Murine models have demonstrated the protective role of cross-reactive CTL against influenza A virus antigenic drift. We tested the ability of an adjuvanted carrier (Iscomatrix) to help human antigen-presenting cells present formalin-killed influenza vaccine to human CD8(+) CTL clones in vitro and in vaccinated humans. The results of a randomized, double-blind, controlled clinical study demonstrate that a single dose of a vaccine formulated into Iscom particles increased influenza A virus specific CTL memory in 50-60% of recipients, compared to 5% of the recipients of the standard influenza vaccine. PMID- 10388650 TI - The evolution of porcine reproductive and respiratory syndrome virus: quasispecies and emergence of a virus subpopulation during infection of pigs with VR-2332. AB - GP5, the principal envelope glycoprotein of porcine reproductive and respiratory syndrome virus (PRRSV), contains a hypervariable region within the ectodomain which is responsible for generating diversity in field isolates. The purpose of this study was to gain insight into the possible origin of this diversity by following GP5 sequence changes in pigs exposed to PRRSV strain VR-2332 in utero. A region of the PRRS virus genome containing portions of ORF4 and ORF5 was amplified directly from tissues of infected pigs from birth to 132 days of age. We observed the emergence of a new PRRSV population, identified by a single nucleotide change in the ectodomain. The Asp to Asn change at amino acid 34 was also found as a minor component in pigs that expressed the wild-type sequence. The results from this study suggest that the variability in the ectodomain of ORF5 is the result of positive or negative selection, of which the mechanism remains to be determined. PMID- 10388651 TI - HIV and SIV gp120 binding does not predict coreceptor function. AB - Interaction of HIV and SIV Envelope (Env) proteins with viral coreceptors is a critical step in viral entry. By using a sensitive and specific gp120 binding assay, we have identified a discordance between the ability of a coreceptor to support Env-mediated membrane fusion and high-affinity binding of gp120. Direct binding of gp120 from the dual-tropic HIV-1 strain 89.6 was not detectable for any coreceptor that it uses for fusion, while detectable binding of gp120s from the R5 HIV-1 strains JRFL and CM235 and the SIV strain 239 was not measurable for many CCR5 chimeras and mutants that function efficiently as viral coreceptors. In comparison, binding of chemokines to these same mutants was highly predictive of their ability to signal. Thus, gp120 is more sensitive than chemokines to perturbations of CCR5 structure. We conclude that while chemokine binding to CCR5 is a good predictor of chemokine receptor function, gp120 binding does not always predict coreceptor function. PMID- 10388652 TI - Isolation and characterization of an oligomerization-negative mutant of HIV-1 integrase. AB - The yeast two-hybrid method was used to screen mutagenized DNAs to isolate a variant of the human immunodeficiency virus type 1 integrase (IN) that does not interact with the wild-type IN. The responsible mutation, leading to a single amino acid change (V260E) in the C-terminal domain of IN, blocks IN-IN multimerization but has only small effect on binding to a host interacting protein, INI1 (hSNF5). Binding studies in vitro confirmed the defect in multimerization of the mutant IN. Biochemical analyses of the mutant IN enzyme expressed in bacteria detected only subtle changes in its properties, suggesting that the yeast system is a sensitive reporter of correct IN conformation. Mutant virus carrying the V260E substitution was blocked in replication at the time of DNA integration, consistent with IN multimerization being important for its activity in vivo. PMID- 10388653 TI - Posttranscriptional regulation of US11 in cells infected with a herpes simplex virus 1 recombinant lacking both 222-bp domains containing S-component origins of DNA synthesis. AB - The US11 gene of herpes simplex virus 1 maps in the unique sequences of the short component of the HSV-1(F) genome approximately 775 bp from the center of the DNA replication origin (OriS) and encodes a virion protein which binds RNA in sequence- and conformation-specific fashion, negatively regulates the accumulation of a prematurely terminated transcript of UL34, associates in the infected cell with the 60S ribosomal subunit, and, late in infection, accumulates in nucleoli. We report the following: (i) Deletion of a 222-bp sequence including OriS (DeltaOriS) negatively affected the accumulation of the US11 protein without decreasing the accumulation of the US11 transcript. (ii) The defect, observed at all times after infection, was multiplicity independent, was unrelated to US11 protein stability, and apparently resulted from a cis-acting element since a coinfecting virus was unable to complement the DeltaOriS virus. (iii) Transcription from the US11 promoter initiated from three sites on the DeltaOriS virus. Transcripts initiated from two of the three initation sites accumulated similarly in cells infected with the DeltaOriS virus or wild-type parent virus. The low-abundance transcript initiating from the third site was apparently unique to the DeltaOriS virus but was not expected to alter the coding capacity of the mRNA. (iv) Infected cells accumulated RNA derived by antisense transcription of the genome domain containing the US11 gene. One transcript accumulated in larger amounts in cells infected with the DeltaOriS virus than in cells infected with parent or repaired virus. PMID- 10388654 TI - Distinct export pathway utilized by the hepatitis B virus posttranscriptional regulatory element. AB - The posttranscriptional regulatory element (PRE) of hepatitis B virus is an RNA element important for the export of viral mRNA from the nucleus to the cytoplasm. The cellular export pathway utilized by the PRE is controversial. We present data showing that PRE-dependent export is blocked by vesicular stomatitis virus matrix protein, an inhibitor of all cellular RNA export other than tRNA export. It is also blocked by a mutated form of Ran-binding protein 1, which blocks export mediated by the human immunodeficiency virus Rev and Rev-response element (RRE) but not export mediated by the simian retrovirus constitutive transport element (CTE). On the other hand, PRE-dependent export is not blocked by either TAgRex or leptomycin B, two agents that prevent Rev/RRE-mediated export. Therefore, PRE appears to utilize an export pathway different from that of Rev/RRE or CTE. PMID- 10388655 TI - The human papillomavirus E7 oncoprotein abrogates signaling mediated by interferon-alpha. AB - Greater than 95% of all cervical carcinomas have been found to be associated with "high-risk" human papillomavirus (mainly types 16 and 18) infections, with the viral E6 and E7 oncoproteins essential for neoplastic development and maintenance. Interferon-alpha (IFNalpha) is used in the treatment of HPV infections yet both in vivo and in vitro data suggest that the virus has developed mechanisms to avoid the effects of interferon. Here we show that the HPV16 E7 oncoprotein is able to inhibit the induction of IFNalpha-inducible genes but has no effect of IFNgamma-inducible genes. Expression of E7 correlates with the loss of formation of the interferon-stimulated gene factor 3 (ISGF3) transcription complex. Moreover, in the presence of E7, p48, the DNA-binding component of ISGF3, was unable to translocate to the nucleus upon IFNalpha stimulation. A direct protein-protein interaction was identified between E7 and p48 with the site of interaction within E7 defined as the region between amino acids 17-37, a domain that includes the binding site for the retinoblastoma protein, pRb. These results suggest that HPV, via E7, targets p48, resulting in the loss of IFNalpha-mediated signal transduction and may provide a means by which HPV can avoid the innate immune system. PMID- 10388656 TI - Expression and detection of macrophage-tropic HIV-1 gp120 in the brain using conformation-dependent antibodies. AB - HIV-1 envelope proteins gp120 and gp41 are likely to play a role in the pathogenesis of HIV-associated neurocognitive disorders. While detection of gp120 in HIV-infected cell cultures is easy, it has not yet been possible to identify gp120 in human or animal brains in situ. The difficulty in detecting gp120 could be due to low expression levels of the protein, to the shedding of gp120 from infected macrophages/microglia, or to the use of inappropriate gp-specific antibodies. We addressed these questions by analyzing the subcellular localization, oligomeric structure, and shedding behavior of gp120 from a macrophage-tropic, CCR5-dependent primary isolate, BX08, expressed by a Semliki Forest virus replicon (SFVenvBX08) in vitro. We used the same SFV system injected in vivo into the rat brain in an attempt to detect gp120 in situ. Our results show that gp120/41 is expressed as monomers, dimers, and trimers in cell culture. Immunocytochemical analysis revealed that intracytoplasmic gp120 can be recognized by an anti-V3 antibody, whereas gp120 at the plasma membrane is detected exclusively by a conformation-dependent antibody. In the rat brain, the SFV vector allows gene expression in neurons from day 3 to day 9 after injection without any apparent brain damage nor reactive astrogliosis. In SFVenvBX08 infected neurons only conformation-dependent antibodies allowed gp120 labeling. These results suggest that previous difficulties in detecting gp120 in brain tissues may be due to the use of antibodies which were unable to recognize gp120 at the plasma membrane. PMID- 10388657 TI - Changes in and discrepancies between cell tropisms and coreceptor uses of human immunodeficiency virus type 1 induced by single point mutations at the V3 tip of the env protein. AB - We examined the effect of amino acid substitutions of the GPGR (glycine-proline glycine-arginine) tip sequence at the V3 domain of the Env protein of human immunodeficiency virus type 1 (HIV-1) on its cell tropism and coreceptor use. We changed the GPGR sequence of a T-cell line (T)- and macrophage (M)-tropic (R5-R3 X4) HIV-1 strain, GUN-1wt, to GA(alanine)GR (the resulting mutant was designated GUN-1/A), GL(leucine)GR (GUN-1/L), GP(proline)GR (GUN-1/P), GR(arginine)GR (GUN 1/R), GS(serine)GR (GUN-1/S), or GT(threonine)GR (GUN-1/T). GUN-1/A, GUN-1/S, and GUN-1/T mutants infected brain-derived cells such as a CD4-transduced glioma cell line, U87/CD4, and a brain-derived primary cell strain, BT-20/N, as well as T cell lines in a CD4-dependent manner, although the plating of these mutants onto macrophages was inhibited. GUN-1/L, GUN-1/P, and GUN-1/R mutants showed both T- and M-tropism, but did not plate onto the brain-derived cells. A CCR3, CCR5, CCR8, or CXCR4 gene was introduced into a CD4-positive glioma cell line, NP 2/CD4, which demonstrated complete resistance to various HIV-1 strains. Not only HIV-1 strains, which were infectious to macrophages, such as GUN-1wt, GUN-1v, GUN 1/L, and GUN-1/P, but also an HIV-1 strain, GUN-1v, which was hardly infectious to macrophages, grew well in NP-2/CD4 cells expressing CCR3 or CCR5. However, the M-tropic GUN-1/R mutant could not efficiently use CCR5 nor CCR3. No point mutants, except GUN-1/L, grew well in NP-2/CD4 cells expressing CCR8. These findings indicate that the cell tropism of HIV-1 to macrophages and brain-derived cells and their use of the coreceptors were markedly, though not always concomitantly, affected by the tip sequence of the V3 domain. PMID- 10388658 TI - Mechanisms of cytokine-mediated inhibition of viral replication. AB - In this report, the role of nitric oxide synthase (NOS) and IL-12 administration in inhibition of vesicular stomatitis virus (VSV) from infected neuroblastoma cells was examined. We previously have shown that cytokine treatment of cells results in the induction of NOS-1, and this is associated with a 2 log inhibition of VSV production. We performed these studies to examine the mechanism by which viral replication is suppressed. Neuroblastoma cells (NB41A3) were treated with either IL-12 or medium and subsequently infected with VSV. Viral protein and mRNA were isolated from these cells, and their levels were measured by Western or Northern blots, respectively. mRNA levels were decreased modestly, but viral proteins were decreased substantially in cells pretreated with IL-12, suggesting that the inhibitory effect of NO is working at the translational level. Cytokine treatment of cells was not associated with oxidative stress. The viral proteins also were nitrosylated. These data suggest that the mechanism of NO inhibition of viral replication occurs through translational interference and posttranslational modifications of viral components. PMID- 10388659 TI - Roles of the three major phosphorylation sites of hepatitis B virus core protein in viral replication. AB - Hepatitis B virus (HBV) core protein is a phosphoprotein. Its three major phosphorylation sites have been identified at the serine residues located at amino acids 157, 164, and 172. In order to investigate the role of core protein phosphorylation in HBV replication, these three serine residues were mutated to alanine to mimic nonphosphorylated serine or to glutamic acid to mimic phosphoserine. The nonphosphorylated core protein analog did not package the HBV pregenomic RNA, and the phosphorylated analog packaged the pregenomic RNA but failed to support viral DNA replication. These results indicate that the core protein phosphorylation may be important for pregenomic RNA packaging and that its dephosphorylation may be important for viral DNA replication. The individual roles of these three major phosphorylation sites in HBV replication were further investigated by being mutated to alanine in different combinations. The results showed that the serine residue at amino acid 157 was not essential for pregenomic RNA packaging, whereas the serine residues at amino acids 164 and 172 were more important. Furthermore, the serine residue at amino acid 157 was not essential for viral DNA replication or viral maturation. PMID- 10388660 TI - Defective nef alleles in a cohort of hemophiliacs with progressing and nonprogressing HIV-1 infection. AB - Deletion of the nef gene results in viral attenuation and confers protection against challenge with wild-type simian immunodeficiency virus in macaques. Regarding HIV-1 infection, a few long-term nonprogressors (LTNP) with nef deletions have been described. In this study, the nef genes of a group of seven LTNP and eight progressors, all belonging to the same cohort of infected hemophiliacs, were analyzed by cloning and sequencing from both virion RNA and peripheral blood mononuclear cell-associated proviral DNA. Defective nef sequences coexisted with full-length nef open reading frames in five of seven LTNP and two of eight progressors. The proportion of disrupted nef sequences within each individual was significantly higher in LTNP (ranging from 10 to 63%) than in progressors (ranging from 9 to 21%) (P = 0.013). Moreover, in-frame small deletions predicting to encode Nef were found in all RNA- and DNA-derived clones from one LTNP and four progressors. A chimeric virus in which the nef gene of NL4.3 was substituted with the nef allele containing the deletion of two alanines at position 49-50 found in two progressors showed a defective replicative capacity compared to NL4.3 virus. In summary, hemophiliacs with either progressing or nonprogressing HIV-1 infection are characterized by the presence of defective nef variants. PMID- 10388661 TI - Modulation of translational efficiency by contextual nucleotides flanking a baculovirus initiator AUG codon. AB - In a previous study of translational regulation of a baculovirus gene, we observed that translation initiated at an unexpectedly high efficiency from an AUG codon found in what was believed to be a poor context (M.-J. Chang and G. W. Blissard, 1997, J. Virol. 71, 7448-7460). In the current study, we examined the roles of nucleotides flanking a baculovirus AUG initiator codon in modulating translation initiation in lepidopteran insect cells. The roles of nucleotides flanking the AcMNPV gp64 initiator codon were examined by site-directed mutagenesis and functional assays in transfected Sf9 cells. To eliminate potential cis-acting sequences and effects, the gp64 initiator context was cloned in-frame with a chloramphenicol acetyl transferase reporter gene and under the control of a heterologous promoter. All possible single-nucleotide substitutions were generated in positions -6 to -1 and +4 to +6, relative to the A of the initiator AUG codon, which was designated +1. Constructs were transfected into lepidopteran cells and translation products were quantified by an enzyme-linked immunosorbent assay procedure. Substitutions of pyrimidines or other nucleotides at the -3 position resulted in little or no detectable effect on translation efficiency. In contrast, specific substitutions at the +4 and +5 positions resulted in approximately 2- to 3-fold increases in translation. Substitution of A in the +4 position resulted in an approximately 3-fold increase in translation, and substitution of any nucleotide for T in the +5 position resulted in approximately 1.9- to 2.8-fold increases. Substitutions at other positions (-6 to -1 and +6) resulted in no detectable increase or decrease in translation efficiency. These experimental results suggest an optimal initiator context of 5' N N N N N N A U G A a/c/g N-3' for efficient translation initiation in lepidopteran cells. Consensus translation initiation contexts were generated from baculovirus genes and lepidopteran genes, then compared with the experimental results from the gp64 initiator context. PMID- 10388662 TI - Down-regulation of the INK4 family of cyclin-dependent kinase inhibitors by tax protein of HTLV-1 through two distinct mechanisms. AB - Tax oncoprotein of human T-cell leukemia virus type 1 (HTLV-1) affects multiple regulatory processes of infected cells through activation and repression of specific transcription and also through modulation of functions of cell cycle regulators. Previously, we found that Tax binds to p16ink4a, a member of the INK4 family of cyclin-dependent kinase inhibitors, and counteracts its inhibitory activity, resulting in cell cycle progression. In this study, we examined the effects of Tax on other members of the INK4 family and found that Tax can bind to p15ink4b similarly to p16ink4a, but not to p18ink4c and p19ink4d. Tax binding to p15ink4b inactivated its function and restored CDK4 kinase activity. Accordingly, Tax-expressing cells became resistant to p15ink4b-mediated growth arrest induced by TGFbeta. On the other hand, expression of p18ink4c was transcriptionally repressed by Tax through the E-box element of the promoter, which may contribute to the marked reduction of p18ink4c mRNA in HTLV-1-infected T-cells. These observations indicate that Tax suppresses the inhibitory activities of INK4 family members through two independent mechanisms: functional inhibition of two INK4 proteins and repression of expression of another INK4 protein. These effects may play roles in HTLV-1-induced deregulation of the cell cycle, possibly promoting cellular transformation. PMID- 10388663 TI - Retrovirus DNA termini bound by integrase communicate in trans for full-site integration in vitro. AB - Integration of linear retrovirus DNA involves the concerted insertion of the viral termini (full-site integration) into the host chromosome. We investigated the interactions that occur between long terminal repeat (LTR) termini bound by avian retrovirus integrase (IN) for full-site integration in vitro. Wild-type (wt) or mutant LTR donors that possess gain-of-function ("G") or loss-of-function ("L") for full-site integration activity were used. G LTR termini are characterized as having significantly higher strand transfer activity than the wt and the L LTR termini. L LTR mutations are classified as partially or extremely defective for strand transfer activity. The L mutations were further classified by their ability to either permit or block the assembly of G or wt LTR termini into nucleoprotein complexes capable of full-site strand transfer. We demonstrated that avian myeloblastosis virus IN bound to G LTR termini increased the incorporation of partially defective L LTR termini into nucleoprotein complexes that were capable of full-site integration. The observed full-site integration activity of these assembled nucleoprotein complexes appeared to be influenced by each individual IN-LTR complex in trans. In contrast, extremely defective L LTR termini exhibited the ability to effectively block the assembly of wt LTR termini into nucleoprotein complexes capable of full-site strand transfer. Data from nonspecific DNA competition experiments suggested that IN had an apparent higher affinity for G LTR donor termini than for partially defective L LTR donor termini as measured by full-site integration activity. However, assembled nucleoprotein complexes containing either two G or two L LTR donors were stable, having a similar half-life of approximately 2 h on ice. The results suggest that LTR termini bound by IN exhibit an allosteric effect to modulate full-site integration in vitro. Similar regulatory controls also appear to exist in vivo between the wt U3 and wt U5 LTR termini in retroviruses as well as purified retrovirus preintegration complexes that promoted full-site integration in vitro. PMID- 10388664 TI - Phosphorylation of the viral nonstructural protein NS1 during MVMp infection of A9 cells. AB - The major nonstructural protein of parvovirus MVMp, NS1, is an 83-kDa nuclear phosphoprotein which exerts a variety of functions during a viral infection. These multiple tasks range from its major involvement in viral DNA amplification and promoter regulation to the cytotoxic action on the host cell. Since these most divergent functions are exerted in an orderly fashion, it has been proposed that NS1 is regulated by posttranslational modifications, in particular phosphorylation. So far it has been shown that the capacity of NS1 for initiation of replication is regulated in vitro by phosphorylation through members of the protein kinase C family, most likely as a result of control of the DNA unwinding activity (J. P. F. Nuesch et al., 1998, J. Virol. 72, 9966-9977). To substantiate these in vitro findings in vivo, we investigated NS1 phosphorylation during an MVMp infection in a natural host cell, A9 fibroblasts, with reference to characteristic features of the virus cycle. The NS1 phosphorylation pattern was found to change throughout the infection, raising the possibility that distinct tasks of NS1 might be achieved through differential phosphorylation of the polypeptide. In addition, we present in vivo evidence that a phosphorylated form of NS1 is able to initiate viral DNA replication and becomes covalently attached to replicated DNA. Moreover, NS1 was found to be phosphorylated in vivo within the helicase domain, showing alignment with at least one phosphopeptide generated by an "activating" kinase in vitro. These data suggest that phosphorylation mediated regulation of NS1 for replicative functions as observed in vitro may also take place during a natural virus infection. PMID- 10388665 TI - In vitro antibody-dependent enhancement assays are insensitive indicators of in vivo vaccine enhancement of equine infectious anemia virus. AB - We have previously demonstrated a high propensity for enhancement of virus replication and disease resulting from experimental immunization of ponies with a baculovirus recombinant envelope (rgp90) vaccine from equine infectious anemia virus (EIAV). The current studies were undertaken to examine the correlation between the observed in vivo vaccine enhancement and in vitro assays for antibody dependent enhancement (ADE) of EIAV replication. Toward this goal an optimized EIAV in vitro enhancement assay was developed using primary equine macrophage cells and used to evaluate the enhancement properties of immune serum taken from rgp90 immunized ponies that displayed various levels of vaccine enhancement after experimental challenge with EIAV. For comparison, we analyzed in parallel immune serum samples from a group of ponies immunized with a viral envelope subunit vaccine (LL-gp) that produced sterile protection from EIAV challenge. The results of these assays demonstrated that the rgp90 immune serum had a greater propensity for in vitro enhancement of EIAV replication than serum from the protected LL-gp immunized ponies; in vitro enhancement levels for the rgp90 immune sera averaged about 1.5, with a maximum enhancement value of about 2.0. While distinguishing between immune serum produced by the rgp90 and LL-gp immunizations, the in vitro enhancement assay failed to reliably correlate with the severity of in vivo enhancement observed among the rgp90 vaccine recipients. Vaccinated ponies that experienced moderate to no disease signs displayed levels of in vitro enhancement similar to those of ponies that experienced severe and fatal enhancement of disease after viral challenge. The observed in vitro enhancement was demonstrated to be dependent on serum immunoglobulin, but independent of complement. These studies demonstrate in the EIAV system that in vitro ADE assays appear to be relatively insensitive indicators of the severity of in vivo enhancement and that relatively low levels of in vitro ADE can be associated with severe to fatal enhancement of virus replication and disease in vivo. These observations suggest that relatively low levels of serum ADE observed in other lentivirus systems, including HIV-1, may have more profound effects on in vivo virus replication and disease than previously recognized. PMID- 10388666 TI - Marked genomic heterogeneity and frequent mixed infection of TT virus demonstrated by PCR with primers from coding and noncoding regions. AB - A nonenveloped, single-stranded, and circular DNA virus designated TT virus (TTV) has been reported in association with hepatitis of unknown etiology. TTV has a wide sequence divergence (approximately 52%), by which it is classified into at least 16 genotypes separated by an evolutionary distance of >0.30. Therefore, the detection of TTV DNA by polymerase chain reaction would be influenced by primers deduced from conserved or divergent regions of the genome. Of the 30 sera from healthy individuals, up to 17% tested positive with primers deduced from coding region, much less frequently than up to 93% testing positive with primers from noncoding region. These differences were not attributable to the sensitivity of detection, because a cloned TTV DNA of genotype 1a was detected sensitively (up to 1 copy per test) with primers deduced from either the coding or the noncoding region of the same genotype. Sera testing positive only with noncoding region primers, or those showing higher titers with noncoding than coding region primers, contained TTV DNA strains with sequence divergence of 47-53% from the TA278 isolate of genotype 1a within the N22 region spanning 222-231 nucleotides. Some of the sera contained two or three TTV DNA strains of distinct genotypes. These results indicate TTV strains with extremely high sequence divergence prevailing in healthy individuals and frequent mixed infection with TTV strains of distinct genotypes. PMID- 10388667 TI - The entire nucleotide sequence of a TT virus isolate from the United States (TUS01): comparison with reported isolates and phylogenetic analysis. AB - A nonenveloped and single-stranded DNA virus designated TT virus (TTV) has been reported from Japan in association with hepatitis of unknown etiology. Very recently, the prototype TTV isolate (TA278) of genotype 1 is proven to have a circular genome with 3852 nucleotides. A TTV isolate (TUS01) was recovered from a blood donor in the United States, and its entire circular nucleotide sequence of 3818 nucleotides was determined. It possessed two open reading frames coding for 761 and 156 amino acids, respectively. TUS01 shared 60.5% of the nucleotide sequence with the TA278 isolate from Japan that was longer by 35 nt. The sequence of the noncoding region of 1203 nt was conserved with a similarity of 83.4%. Sequence preservation was much lower for the two open reading frames; nucleotide and amino acid sequences were 54.8 and 37.0% similar, respectively, for one and 55.5 and 38.8% similar for the other. By comparison of a partial sequence of 222 nucleotides among 239 TTV isolates available from various countries, at least 11 genotypes with sequence divergence of >30% were recognized. TUS01 was deduced to be of genotype 11, which has not been reported before. Conserved sequences in the noncoding region could be used as primers for sensitively detecting TTV DNA by polymerase chain reaction. Divergent sequences in coding regions would be useful as primers for distinguishing various TTV genotypes. PMID- 10388668 TI - Changes of fermentation pathways of fecal microbial communities associated with a drug treatment that increases dietary starch in the human colon. AB - Acarbose inhibits starch digestion in the human small intestine. This increases the amount of starch available for microbial fermentation to acetate, propionate, and butyrate in the colon. Relatively large amounts of butyrate are produced from starch by colonic microbes. Colonic epithelial cells use butyrate as an energy source, and butyrate causes the differentiation of colon cancer cells. In this study we investigated whether colonic fermentation pathways changed during treatment with acarbose. We examined fermentations by fecal suspensions obtained from subjects who participated in an acarbose-placebo crossover trial. After incubation with [1-13C]glucose and 12CO2 or with unlabeled glucose and 13CO2, the distribution of 13C in product C atoms was determined by nuclear magnetic resonance spectrometry and gas chromatography-mass spectrometry. Regardless of the treatment, acetate, propionate, and butyrate were produced from pyruvate formed by the Embden-Meyerhof-Parnas pathway. Considerable amounts of acetate were also formed by the reduction of CO2. Butyrate formation from glucose increased and propionate formation decreased with acarbose treatment. Concomitantly, the amounts of CO2 reduced to acetate were 30% of the total acetate in untreated subjects and 17% of the total acetate in the treated subjects. The acetate, propionate, and butyrate concentrations were 57, 20, and 23% of the total final concentrations, respectively, for the untreated subjects and 57, 13, and 30% of the total final concentrations, respectively, for the treated subjects. PMID- 10388670 TI - Method for detection and enumeration of Cryptosporidium parvum oocysts in feces, manures, and soils. AB - Eight concentration and purification methods were evaluated to determine percentages of recovery of Cryptosporidium parvum oocysts from calf feces. The NaCl flotation method generally resulted in the highest percentages of recovery. Based on the percentages of recovery, the amounts of fecal debris in the final oocyst preparations, the relatively short processing time (<3 h), and the low expense, the NaCl flotation method was chosen for further evaluation. Extraction efficiency was evaluated by using oocyst concentrations of 25, 50, 10(2), 10(3), 10(4), and 10(5) oocysts g of bovine feces-1. The percentages of recovery ranged from 10.8% (25 oocysts g-1) to 17.0% (10(4) oocysts g-1) (r2 = 0.996). A conservative estimate of the detection limit for bovine feces is ca. 30 oocysts g of feces-1. Percentages of recovery were determined for six different types of animal feces (cow, horse, pig, sheep, deer, and chicken feces) at a single oocyst concentration (10(4) oocysts g-1). The percentages of recovery were highest for bovine feces (17. 0%) and lowest for chicken feces (3.2%). Percentages of recovery were determined for bovine manure after 3 to 7 days of storage. The percentages of recovery ranged from 1.9 to 3.5% depending on the oocyst concentration, the time of storage, and the dispersing solution. The percentages of oocyst recovery from soils were evaluated by using different flotation solutions (NaCl, cold sucrose, ZnSO4), different dispersing solutions (Triton X 100, Tween 80, Tris plus Tween 80), different dispersion techniques (magnetic stirring, sonication, blending), and different dispersion times (5, 15, and 30 min). Twenty-five-gram soil samples were used to reduce the spatial variability. The highest percentages of recovery were obtained when we used 50 mM Tris-0.5% Tween 80 as the dispersing solution, dispersion for 15 min by stirring, and saturated NaCl as the flotation solution. The percentages of oocyst recovery from freshly spiked sandy loam, silty clay loam, and clay loam soils were ca. 12 to 18, 8, and 6%, respectively. The theoretical detection limits were ca. 1 to 2 oocysts g of soil-1 depending on the soil type. The percentages of recovery without dispersant (distilled H2O or phosphate-buffered saline) were less than 0.1%, which indicated that oocysts adhere to soil particles. The percentages of recovery decreased with storage time, although the addition of dispersant (Tris Tween 80) before storage appeared to partially prevent adhesion. These data indicate that the NaCl flotation method is suitable for routine detection and enumeration of oocysts from feces, manures, soils, or soil-manure mixtures. PMID- 10388669 TI - An outbreak of nonflocculating catabolic populations caused the breakdown of a phenol-digesting activated-sludge process. AB - Activated sludge was fed phenol as the sole carbon source, and the phenol-loading rate was increased stepwise from 0.5 to 1.0 g liter-1 day-1 and then to 1.5 g liter-1 day-1. After the loading rate was increased to 1.5 g liter-1 day-1, nonflocculating bacteria outgrew the sludge, and the activated-sludge process broke down within 1 week. The bacterial population structure of the activated sludge was analyzed by temperature gradient gel electrophoresis (TGGE) of PCR amplified 16S ribosomal DNA (rDNA) fragments. We found that the population diversity decreased as the phenol-loading rate increased and that two populations (designated populations R6 and R10) predominated in the sludge during the last several days before breakdown. The R6 population was present under the low-phenol loading-rate conditions, while the R10 population was present only after the loading rate was increased to 1.5 g liter-1 day-1. A total of 41 bacterial strains with different repetitive extragenic palindromic sequence PCR patterns were isolated from the activated sludge under different phenol-loading conditions, and the 16S rDNA and gyrB fragments of these strains were PCR amplified and sequenced. Some bacterial isolates could be associated with major TGGE bands by comparing the 16S rDNA sequences. All of the bacterial strains affiliated with the R6 population had almost identical 16S rDNA sequences, while the gyrB phylogenetic analysis divided these strains into two physiologically divergent groups; both of these groups of strains could grow on phenol, while one group (designated the R6F group) flocculated in laboratory media and the other group (the R6T group) did not. A competitive PCR analysis in which specific gyrB sequences were used as the primers showed that a population shift from R6F to R6T occurred following the increase in the phenol-loading rate to 1.5 g liter-1 day 1. The R10 population corresponded to nonflocculating phenol-degrading bacteria. Our results suggest that an outbreak of nonflocculating catabolic populations caused the breakdown of the activated-sludge process. This study also demonstrated the usefulness of gyrB-targeted fine population analyses in microbial ecology. PMID- 10388671 TI - Production of monoclonal antibodies to Listeria monocytogenes and their application to determine the virulence of isolates from channel catfish. AB - We produced monoclonal antibodies (MAbs) to the extracellular proteins of Listeria monocytogenes EGD grown in Chelex-treated improved minimal medium. Ten of the positive hybridomas generated were chosen for further characterization. Seven of the MAbs reacted with a protein having a molecular mass of 60 kDa. These MAbs inhibited listeriolysin (LLO)-mediated hemolysis, and two of them were specific for LLO and none of the other thiol-activated toxins tested. In an enzyme-linked immunosorbent assay and Western blot analysis, five of the anti-LLO MAbs reacted with ivanolysin from Listeria ivanovii. Three of the 10 MAbs reacted with a 29-kDa protein on Western blots and neutralized the phosphatidylcholine specific phospholipase C (PC-PLC) activity of L. monocytogenes. These three anti PC-PLC MAbs did not react with phospholipases from five different gram-positive bacteria. However, the anti-PC-PLC MAbs recognized a 27-kDa extracellular protein from L. ivanovii and neutralized sphingomyelinase activity in a hemolysis test that demonstrates the antigenic relatedness of listerial phospholipases. These data indicate that listerial thiol-activated toxins possess species-specific epitopes and share group-specific epitopes. This is the first description of MAbs that neutralize listerial PC-PLC, and the data suggest that there is antigenic similarity between L. monocytogenes PC-PLC and L. ivanovii sphingomyelinase. The reactions of the MAbs with catfish isolates of L. monocytogenes suggested that some of the isolates examined lack the LLO and/or PC-PLC required for pathogenicity. The MAbs described here differentiated some catfish isolates from previously described type strain-pathogenic isolates and could be useful for detecting and determining the virulence of L. monocytogenes in food and clinical samples and for detecting L. ivanovii in veterinary clinical samples. PMID- 10388672 TI - Cloning and characterization of a Rhizobium leguminosarum gene encoding a bacteriocin with similarities to RTX toxins. AB - A 3-kb region containing the determinant for bacteriocin activity from Rhizobium leguminosarum 248 was isolated and characterized by Tn5 insertional mutagenesis and DNA sequencing. Southern hybridizations showed that this bacteriocin was encoded on the plasmid pRL1JI and that homologous loci were not found in other unrelated R. leguminosarum strains. Tn5 insertional mutagenesis showed that mutations in the C-terminal half of the bacteriocin open reading frame apparently did not abolish bacteriocin activity. Analysis of the deduced amino acid sequence revealed that, similarly to RTX proteins (such as hemolysin and leukotoxin), this protein contains a characteristic nonapeptide repeated up to 18 times within the protein. In addition, a novel 19- to 25-amino-acid motif that occurred every 130 amino acids was detected. Bacteriocin bioactivity was correlated with the presence of a protein of approximately 100 kDa in the culture supernatants, and the bacteriocin bioactivity demonstrated a calcium dependence in both R. leguminosarum and Sinorhizobium meliloti. A mutant of strain 248 unable to produce this bacteriocin was found to have a statistically significant reduction in competitiveness for nodule occupancy compared to two test strains in coinoculation assays. However, this strain was unable to compete any more successfully with a third test strain, 3841, than was wild-type 248. PMID- 10388673 TI - Stress tolerance in doughs of Saccharomyces cerevisiae trehalase mutants derived from commercial Baker's yeast. AB - Accumulation of trehalose is widely believed to be a critical determinant in improving the stress tolerance of the yeast Saccharomyces cerevisiae, which is commonly used in commercial bread dough. To retain the accumulation of trehalose in yeast cells, we constructed, for the first time, diploid homozygous neutral trehalase mutants (Deltanth1), acid trehalase mutants (Deltaath1), and double mutants (Deltanth1 ath1) by using commercial baker's yeast strains as the parent strains and the gene disruption method. During fermentation in a liquid fermentation medium, degradation of intracellular trehalose was inhibited with all of the trehalase mutants. The gassing power of frozen doughs made with these mutants was greater than the gassing power of doughs made with the parent strains. The Deltanth1 and Deltaath1 strains also exhibited higher levels of tolerance of dry conditions than the parent strains exhibited; however, the Deltanth1 ath1 strain exhibited lower tolerance of dry conditions than the parent strain exhibited. The improved freeze tolerance exhibited by all of the trehalase mutants may make these strains useful in frozen dough. PMID- 10388674 TI - Quantification of chemotaxis to naphthalene by Pseudomonas putida G7. AB - The capillary assay was used to quantify the chemotactic response of Pseudomonas putida G7 to naphthalene. Experiments were conducted in which the cell concentration in the assay chamber, the naphthalene concentration in the capillary, or the incubation time was varied. Data from these experiments were evaluated with a model that accounted for the effect of diffusion on the distribution of substrate and the transport of cells from the chamber through the capillary orifice. By fitting a numerical solution of this model to the data, it was possible to determine the chemotactic sensitivity coefficient, chi0. The mean of the best-fit values for chi0 from the three types of experiments was 7.2 x 10( 5) cm2/s. A less computationally intensive model based on earlier approaches that ignore cell transport in the chamber resulted in chi0 values that were approximately three times higher. The models evaluated in the present study could simulate the results of capillary assays only at low chamber cell concentrations, for which the effect of consumption on the distribution of substrate was negligible. Results from this work suggest that it is possible to use the capillary assay to quantify taxis towards environmentally relevant chemoeffectors that have low aqueous solubility. PMID- 10388675 TI - Influence of kernel age on fumonisin B1 production in maize by Fusarium moniliforme. AB - Production of fumonisins by Fusarium moniliforme on naturally infected maize ears is an important food safety concern due to the toxic nature of this class of mycotoxins. Assessing the potential risk of fumonisin production in developing maize ears prior to harvest requires an understanding of the regulation of toxin biosynthesis during kernel maturation. We investigated the developmental-stage dependent relationship between maize kernels and fumonisin B1 production by using kernels collected at the blister (R2), milk (R3), dough (R4), and dent (R5) stages following inoculation in culture at their respective field moisture contents with F. moniliforme. Highly significant differences (P 6)-beta-D fructofuranan or levan, the first example of levan synthesis by a Lactobacillus species. Strain LB 121 possesses glucansucrase and levansucrase enzymes that occur in a cell-associated and a cell-free state after growth on sucrose, raffinose, or maltose but remain cell associated during growth on glucose. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of sucrose culture supernatants, followed by staining of gels for polysaccharide synthesizing activity with sucrose as a substrate, revealed the presence of a single glucansucrase protein of 146 kDa. Growth of strain LB 121 in chemostat cultures resulted in rapid accumulation of spontaneous exopolysaccharide-negative mutants that had lost both glucansucrase and levansucrase (e.g., strain K-24). Mutants lacking all levansucrase activity specifically emerged following a pH shiftdown (e.g., strain 35-5). Strain 35-5 still possessed glucansucrase and synthesized wild-type glucan. PMID- 10388695 TI - Purification, characterization, gene cloning, sequencing, and overexpression of aminopeptidase N from Streptococcus thermophilus A. AB - The general aminopeptidase PepN from Streptococcus thermophilus A was purified to protein homogeneity by hydroxyapatite, anion-exchange, and gel filtration chromatographies. The PepN enzyme was estimated to be a monomer of 95 kDa, with maximal activity on N-Lys-7-amino-4-methylcoumarin at pH 7 and 37 degrees C. It was strongly inhibited by metal chelating agents, suggesting that it is a metallopeptidase. The activity was greatly restored by the bivalent cations Co2+, Zn2+, and Mn2+. Except for proline, glycine, and acidic amino acid residues, PepN has a broad specificity on the N-terminal amino acid of small peptides, but no significant endopeptidase activity has been detected. The N-terminal and short internal amino acid sequences of purified PepN were determined. By using synthetic primers and a battery of PCR techniques, the pepN gene was amplified, subcloned, and further sequenced, revealing an open reading frame of 2,541 nucleotides encoding a protein of 847 amino acids with a molecular weight of 96,252. Amino acid sequence analysis of the pepN gene translation product shows high homology with other PepN enzymes from lactic acid bacteria and exhibits the signature sequence of the zinc metallopeptidase family. The pepN gene was cloned in a T7 promoter-based expression plasmid and the 452-fold overproduced PepN enzyme was purified to homogeneity from the periplasmic extract of the host Escherichia coli strain. The overproduced enzyme showed the same catalytic characteristics as the wild-type enzyme. PMID- 10388697 TI - Factors influencing in vitro killing of bacteria by hemocytes of the eastern oyster (Crassostrea virginica). AB - A tetrazolium dye reduction assay was used to study factors governing the killing of bacteria by oyster hemocytes. In vitro tests were performed on bacterial strains by using hemocytes from oysters collected from the same location in winter and summer. Vibrio parahaemolyticus strains, altered in motility or colonial morphology (opaque and translucent), and Listeria monocytogenes mutants lacking catalase, superoxide dismutase, hemolysin, and phospholipase activities were examined in winter and summer. Vibrio vulnificus strains, opaque and translucent (with and without capsules), were examined only in summer. Among V. parahaemolyticus and L. monocytogenes, significantly (P < 0.05) higher levels of killing by hemocytes were observed in summer than in winter. L. monocytogenes was more resistant than V. parahaemolyticus or V. vulnificus to the bactericidal activity of hemocytes. In winter, both translucent strains of V. parahaemolyticus showed significantly (P < 0.05) higher susceptibility to killing by hemocytes than did the wild-type opaque strain. In summer, only one of the V. parahaemolyticus translucent strains showed significantly (P < 0.05) higher susceptibility to killing by hemocytes than did the wild-type opaque strain. No significant differences (P > 0.05) in killing by hemocytes were observed between opaque (encapsulated) and translucent (nonencapsulated) pairs of V. vulnificus. Activities of 19 hydrolytic enzymes were measured in oyster hemolymph collected in winter and summer. Only one enzyme, esterase (C4), showed a seasonal difference in activity (higher in winter than in summer). These results suggest that differences existed between bacterial genera in their ability to evade killing by oyster hemocytes, that a trait(s) associated with the opaque phenotype may have enabled V. parahaemolyticus to evade killing by the oyster's cellular defense, and that bactericidal activity of hemocytes was greater in summer than in winter. PMID- 10388698 TI - Production of Cry11A and Cry11Ba toxins in Bacillus sphaericus confers toxicity towards Aedes aegypti and resistant Culex populations. AB - Cry11A from Bacillus thuringiensis subsp. israelensis and Cry11Ba from Bacillus thuringiensis subsp. jegathesan were introduced, separately and in combination, into the chromosome of Bacillus sphaericus 2297 by in vivo recombination. Two loci on the B. sphaericus chromosome were chosen as target sites for recombination: the binary toxin locus and the gene encoding the 36-kDa protease that may be responsible for the cleavage of the Mtx protein. Disruption of the protease gene did not increase the larvicidal activity of the recombinant strain against Aedes aegypti and Culex pipiens. Synthesis of the Cry11A and Cry11Ba toxins made the recombinant strains toxic to A. aegypti larvae to which the parental strain was not toxic. The strain containing Cry11Ba was more toxic than strains containing the added Cry11A or both Cry11A and Cry11Ba. The production of the two toxins together with the binary toxin did not significantly increase the toxicity of the recombinant strain to susceptible C. pipiens larvae. However, the production of Cry11A and/or Cry11Ba partially overcame the resistance of C. pipiens SPHAE and Culex quinquefasciatus GeoR to B. sphaericus strain 2297. PMID- 10388699 TI - High-level production of human leptin by fed-batch cultivation of recombinant Escherichia coli and its purification. AB - Human leptin is a 16-kDa (146-amino-acid) protein that is secreted from adipocytes and influences body weight homeostasis. In order to obtain high-level production of leptin, the human obese gene coding for leptin was expressed in Escherichia coli BL21(DE3) under the strong inducible T7 promoter. The recombinant leptin was produced as inclusion bodies in E. coli, and the recombinant leptin content was as high as 54% of the total protein content. For production of recombinant human leptin in large amounts, pH-stat fed-batch cultures were grown. Expression of leptin was induced at three different cell optical densities at 600 nm (OD600), 30, 90, and 140. When cells were induced at an OD600 of 90, the amount of leptin produced was 9.7 g/liter (37% of the total protein). After simple purification steps consisting of inclusion body isolation, denaturation and refolding, and anion-exchange chromatography, 144.9 mg of leptin that was more than 90% pure was obtained from a 50-ml culture, and the recovery yield was 41.1%. PMID- 10388701 TI - A sensitive nested reverse transcriptase PCR assay to detect viable cells of the fish pathogen Renibacterium salmoninarum in Atlantic salmon (Salmo salar L.). AB - A nested reverse transcriptase (RT) PCR assay detected mRNA of the salmonid pathogen Renibacterium salmoninarum in samples of RNA extracts of between 1 and 10 cells. Total RNA was extracted from cultured bacteria, Atlantic salmon (Salmo salar L.) kidney tissue and ovarian fluid seeded with the pathogen, and kidney tissue from both experimentally challenged and commercially raised fish. Following DNase treatment, extracted RNA was amplified by both RT PCR and PCR by using primers specific for the gene encoding the major protein antigen of R. salmoninarum. A 349-bp amplicon was detected by polyacrylamide gel electrophoresis and silver stain. Inactivation of cultured bacteria by rifampin or erythromycin produced a loss of nested RT PCR mRNA detection corresponding to a loss of bacterial cell viability determined from plate counts but no loss of DNA detection by PCR. In subclinically diseased fish, nested RT PCR identified similar levels of infected fish as determined by viable pathogen culture. Higher percentages of fish testing positive were generated by PCR, particularly in samples from fish previously subjected to antibiotic chemotherapy where 93% were PCR positive, but only 7% were nested RT PCR and culture positive. PCR can generate false-positive data from amplification of target DNA from nonviable pathogen cells. Therefore, nested RT PCR may prove useful for monitoring cultured Atlantic salmon for the presence of viable R. salmoninarum within a useful time frame, particularly samples from broodstock where antibiotic chemotherapy is used prior to spawning to reduce vertical pathogen transmission. PMID- 10388700 TI - The groESL chaperone operon of Lactobacillus johnsonii. AB - The Lactobacillus johnsonii VPI 11088 groESL operon was localized on the chromosome near the insertion element IS1223. The operon was initially cloned as a series of three overlapping PCR fragments, which were sequenced and used to design primers to amplify the entire operon. The amplified fragment was used as a probe to recover the chromosomal copy of the groESL operon from a partial library of L. johnsonii VPI 11088 (NCK88) DNA, cloned in the shuttle vector pTRKH2. The 2,253-bp groESL fragment contained three putative open reading frames, two of which encoded the ubiquitous GroES and GroEL chaperone proteins. Analysis of the groESL promoter region revealed three transcription initiation sites, as well as three sets of inverted repeats (IR) positioned between the transcription and translation start sites. Two of the three IR sets bore significant homology to the CIRCE elements, implicated in negative regulation of the heat shock response in many bacteria. Northern analysis and primer extension revealed that multiple temperature-sensitive promoters preceded the groESL chaperone operon, suggesting that stress protein production in L. johnsonii is strongly regulated. Maximum groESL transcription activity was observed following a shift to 55 degrees C, and a 15 to 30-min exposure of log-phase cells to this temperature increased the recovery of freeze-thawed L. johnsonii VPI 11088. These results suggest that a brief, preconditioning heat shock can be used to trigger increased chaperone production and provide significant cross-protection from the stresses imposed during the production of frozen culture concentrates. PMID- 10388702 TI - Survival of low-pH stress by Escherichia coli O157:H7: correlation between alterations in the cell envelope and increased acid tolerance. AB - Survival of a nontoxigenic isolate of Escherichia coli O157:H7 at low pH (pH 3.0) was examined over prolonged time periods for each of three population types: exponential-phase cells, stationary-phase cells, and acid-adapted exponential phase cells. In each population, approximately 5 x 10(4) CFU ml-1 were detected after a 24-h incubation at pH 3.0. Even after 3 days at pH 3.0, significant numbers of survivors from each of the three populations could be detected. The high level of acid tolerance exhibited by these survivors was found to be quickly lost once they were transferred to conditions which permitted growth to resume, indicating that they were not mutants. Proton flux measurements on the three populations of cells revealed that the initial rates of viability loss at pH 3.0 correlated well with net proton accumulation. Cells showing a high initial rate of viability loss (exponential-phase cells) accumulated protons at the highest rate, whereas resistant populations (adapted or stationary-phase cells) accumulated protons only slowly. Differences in the protein composition of the cell envelope between the three populations were studied by two-dimensional polyacrylamide gel electrophoresis. Complex differences in the pattern of proteins expressed by each population were uncovered. The implications of these findings are discussed in the context of a possible model accounting for acid tolerance in this important food-borne pathogen. PMID- 10388703 TI - Microbial communities associated with anaerobic benzene degradation in a petroleum-contaminated aquifer. AB - Microbial community composition associated with benzene oxidation under in situ Fe(III)-reducing conditions in a petroleum-contaminated aquifer located in Bemidji, Minn., was investigated. Community structure associated with benzene degradation was compared to sediment communities that did not anaerobically oxidize benzene which were obtained from two adjacent Fe(III)-reducing sites and from methanogenic and uncontaminated zones. Denaturing gradient gel electrophoresis of 16S rDNA sequences amplified with bacterial or Geobacteraceae specific primers indicated significant differences in the composition of the microbial communities at the different sites. Most notable was a selective enrichment of microorganisms in the Geobacter cluster seen in the benzene degrading sediments. This finding was in accordance with phospholipid fatty acid analysis and most-probable-number-PCR enumeration, which indicated that members of the family Geobacteraceae were more numerous in these sediments. A benzene oxidizing Fe(III)-reducing enrichment culture was established from benzene degrading sediments and contained an organism closely related to the uncultivated Geobacter spp. This genus contains the only known organisms that can oxidize aromatic compounds with the reduction of Fe(III). Sequences closely related to the Fe(III) reducer Geothrix fermentans and the aerobe Variovorax paradoxus were also amplified from the benzene-degrading enrichment and were present in the benzene-degrading sediments. However, neither G. fermentans nor V. paradoxus is known to oxidize aromatic compounds with the reduction of Fe(III), and there was no apparent enrichment of these organisms in the benzene-degrading sediments. These results suggest that Geobacter spp. play an important role in the anaerobic oxidation of benzene in the Bemidji aquifer and that molecular community analysis may be a powerful tool for predicting a site's capacity for anaerobic benzene degradation. PMID- 10388704 TI - Composition of toluene-degrading microbial communities from soil at different concentrations of toluene. AB - Toluene-degrading bacteria were isolated from hydrocarbon-contaminated soil by incubating liquid enrichment cultures and agar plate cultures in desiccators in which the vapor pressure of toluene was controlled by dilution with vacuum pump oil. Incubation in desiccators equilibrated with either 100, 10, or 1% (wt/wt) toluene in vacuum pump oil and testing for genomic cross-hybridization resulted in four genomically distinct strains (standards) capable of growth on toluene (strains Cstd1, Cstd2, Cstd5, and Cstd7). The optimal toluene concentrations for growth of these standards on plating media differed considerably. Cstd1 grew best in an atmosphere equilibrated with 0.1% (wt/wt) toluene, but Cstd5 failed to grow in this atmosphere. Conversely, Cstd5 grew well in the presence of 10% (wt/wt) toluene, which inhibited growth of Cstd1. 16S ribosomal DNA sequencing and cross hybridization analysis indicated that both Cstd1 and Cstd5 are members of the genus Pseudomonas. An analysis of the microbial communities in soil samples that were incubated with 10% (wt/wt) toluene with reverse sample genome probing indicated that Pseudomonas strain Cstd5 was the dominant community member. However, incubation of soil samples with 0.1% (wt/wt) toluene resulted in a community that was dominated by Pseudomonas strain Q7, a toluene degrader that has been described previously (Y. Shen, L. G. Stehmeier, and G. Voordouw, Appl. Environ. Microbiol. 64:637-645, 1998). Q7 was not able to grow by itself in an atmosphere equilibrated with 0.1% (wt/wt) toluene but grew efficiently in coculture with Cstd1, suggesting that toluene or metabolic derivatives of toluene were transferred from Cstd1 to Q7. PMID- 10388705 TI - Purification and characterization of a secreted laccase of Gaeumannomyces graminis var. tritici. AB - We purified a secreted fungal laccase from filtrates of Gaeumannomyces graminis var. tritici cultures induced with copper and xylidine. The active protein had an apparent molecular mass of 190 kDa and yielded subunits with molecular masses of 60 kDa when denatured and deglycosylated. This laccase had a pI of 5.6 and an optimal pH of 4.5 with 2,6-dimethoxyphenol as its substrate. Like other, previously purified laccases, this one contained several copper atoms in each subunit, as determined by inductively coupled plasma spectroscopy. The active enzyme catalyzed the oxidation of 2, 6-dimethoxyphenol (Km = 2.6 x 10(-5) +/- 7 x 10(-6) M), catechol (Km = 2.5 x 10(-4) +/- 1 x 10(-5) M), pyrogallol (Km = 3.1 x 10(-4) +/- 4 x 10(-5) M), and guaiacol (Km = 5.1 x 10(-4) +/- 2 x 10(-5) M). In addition, the laccase catalyzed the polymerization of 1, 8-dihydroxynaphthalene, a natural fungal melanin precursor, into a high-molecular-weight melanin and catalyzed the oxidation, or decolorization, of the dye poly B-411, a lignin-like polymer. These findings indicate that this laccase may be involved in melanin polymerization in this phytopathogen's hyphae and/or in lignin depolymerization in its infected plant host. PMID- 10388706 TI - Isolation and characterization of proteolytic ruminal bacteria from sheep and goats fed the tannin-containing shrub legume Calliandra calothyrsus. AB - Tannins in forages complex with protein and reduce the availability of nitrogen to ruminants. Ruminal bacteria that ferment protein or peptides in the presence of tannins may benefit digestion of these diets. Bacteria from the rumina of sheep and goats fed Calliandra calothyrsus (3.6% N and 6% condensed tannin) were isolated on proteinaceous agar medium overlaid with either condensed (calliandra tannin) or hydrolyzable (tannic acid) tannin. Fifteen genotypes were identified, based on 16S ribosomal DNA-restriction fragment length polymorphism analysis, and all were proteolytic and fermented peptides to ammonia. Ten of the isolates grew to high optical density (OD) on carbohydrates (glucose, cellobiose, xylose, xylan, starch, and maltose), while the other isolates did not utilize or had low growth on these substrates. In pure culture, representative isolates were unable to ferment protein that was present in calliandra or had been complexed with tannin. One isolate, Lp1284, had high protease activity (80 U), a high specific growth rate (0.28), and a high rate of ammonia production (734 nmol/min/ml/OD unit) on Casamino Acids and Trypticase Peptone. Phylogenetic analysis of the 16S ribosomal DNA sequence showed that Lp1284 was related (97. 6%) to Clostridium botulinum NCTC 7273. Purified plant protein and casein also supported growth of Lp1284 and were fermented to ammonia. This is the first report of a proteolytic, ammonia-hyperproducing bacterium from the rumen. In conclusion, a diverse group of proteolytic and peptidolytic bacteria were present in the rumen, but the isolates could not digest protein that was complexed with condensed tannin. PMID- 10388707 TI - Photosynthetic bradyrhizobia from Aeschynomene spp. are specific to stem nodulated species and form a separate 16S ribosomal DNA restriction fragment length polymorphism group. AB - We obtained nine bacterial isolates from root or collar nodules of the non-stem nodulated Aeschynomene species A. elaphroxylon, A. uniflora, or A. schimperi and 69 root or stem nodule isolates from the stem-nodulated Aeschynomene species A. afraspera, A. ciliata, A. indica, A. nilotica, A. sensitiva, and A. tambacoundensis from various places in Senegal. These isolates, together with 45 previous isolates from various Aeschynomene species, were studied for host specific nodulation within the genus Aeschynomene, also revisiting cross inoculation groups described previously by D. Alazard (Appl. Environ. Microbiol. 50:732-734, 1985). The whole collection of Aeschynomene nodule isolates was screened for synthesis of photosynthetic pigments by spectrometry, high-pressure liquid chromatography, and thin-layer chromatography analyses. The presence of puf genes in photosynthetic Aeschynomene isolates was evidenced both by Southern hybridization with a Rhodobacter capsulatus photosynthetic gene probe and by DNA amplification with primers defined from photosynthetic genes. In addition, amplified 16S ribosomal DNA restriction analysis was performed on 45 Aeschynomene isolates, including strain BTAi1, and 19 reference strains from Bradyrhizobium japonicum, Bradyrhizobium elkanii, and other Bradyrhizobium sp. strains of uncertain taxonomic positions. The 16S rRNA gene sequence of the photosynthetic strain ORS278 (LMG 12187) was determined and compared to sequences from databases. Our main conclusion is that photosynthetic Aeschynomene nodule isolates share the ability to nodulate particular stem-nodulated species and form a separate subbranch on the Bradyrhizobium rRNA lineage, distinct from B. japonicum and B. elkanii. PMID- 10388708 TI - Visualization and modelling of the thermal inactivation of bacteria in a model food. AB - A large number of incidents of food poisoning have been linked to undercooked meat products. The use of mathematical modelling to describe heat transfer within foods, combined with data describing bacterial thermal inactivation, may prove useful in developing safer food products while minimizing thermal overprocessing. To examine this approach, cylindrical agar blocks containing immobilized bacteria (Salmonella typhimurium and Brochothrix thermosphacta) were used as a model system in this study. The agar cylinders were subjected to external conduction heating by immersion in a water bath. They were then incubated, sliced open, and examined by image analysis techniques for regions of no bacterial growth. A finite-difference scheme was used to model thermal conduction and the consequent bacterial inactivation. Bacterial inactivation rates were modelled with values for the time required to reduce bacterial number by 90% (D) and the temperature increase required to reduce D by 90% taken from the literature. Model simulation results agreed well with experimental results for both bacteria, demonstrating the utility of the technique. PMID- 10388709 TI - Expression of the Escherichia coli catabolic threonine dehydratase in Corynebacterium glutamicum and its effect on isoleucine production. AB - The catabolic or biodegradative threonine dehydratase (E.C. 4.2.1. 16) of Escherichia coli is an isoleucine feedback-resistant enzyme that catalyzes the degradation of threonine to alpha-ketobutyrate, the first reaction of the isoleucine pathway. We cloned and expressed this enzyme in Corynebacterium glutamicum. We found that while the native threonine dehydratase of C. glutamicum was totally inhibited by 15 mM isoleucine, the heterologous catabolic threonine dehydratase expressed in the same strain was much less sensitive to isoleucine; i.e., it retained 60% of its original activity even in the presence of 200 mM isoleucine. To determine whether expressing the catabolic threonine dehydratase (encoded by the tdcB gene) provided any benefit for isoleucine production compared to the native enzyme (encoded by the ilvA gene), fermentations were performed with the wild-type strain, an ilvA-overexpressing strain, and a tdcB expressing strain. By expressing the heterologous catabolic threonine dehydratase in C. glutamicum, we were able to increase the production of isoleucine 50-fold, whereas overexpression of the native threonine dehydratase resulted in only a fourfold increase in isoleucine production. Carbon balance data showed that when just one enzyme, the catabolic threonine dehydratase, was overexpressed, 70% of the carbon available for the lysine pathway was redirected into the isoleucine pathway. PMID- 10388710 TI - Reductive dechlorination of chlorinated ethenes and 1, 2-dichloroethane by "Dehalococcoides ethenogenes" 195. AB - "Dehalococcoides ethenogenes" 195 can reductively dechlorinate tetrachloroethene (PCE) completely to ethene (ETH). When PCE-grown strain 195 was transferred (2% [vol/vol] inoculum) into growth medium amended with trichloroethene (TCE), cis dichloroethene (DCE), 1,1-DCE, or 1,2-dichloroethane (DCA) as an electron acceptor, these chlorinated compounds were consumed at increasing rates over time, which indicated that growth occurred. Moreover, the number of cells increased when TCE, 1,1-DCE, or DCA was present. PCE, TCE, 1,1-DCE, and cis-DCE were converted mainly to vinyl chloride (VC) and then to ETH, while DCA was converted to ca. 99% ETH and 1% VC. cis-DCE was used at lower rates than PCE, TCE, 1,1-DCE, or DCA was used. When PCE-grown cultures were transferred to media containing VC or trans-DCE, products accumulated slowly, and there was no increase in the rate, which indicated that these two compounds did not support growth. When the intermediates in PCE dechlorination by strain 195 were monitored, TCE was detected first, followed by cis-DCE. After a lag, VC, 1,1-DCE, and trans-DCE accumulated, which is consistent with the hypothesis that cis-DCE is the precursor of these compounds. Both cis-DCE and 1,1-DCE were eventually consumed, and both of these compounds could be considered intermediates in PCE dechlorination, whereas the small amount of trans-DCE that was produced persisted. Cultures grown on TCE, 1,1-DCE, or DCA could immediately dechlorinate PCE, which indicated that PCE reductive dehalogenase activity was constitutive when these electron acceptors were used. PMID- 10388711 TI - Effect of growth temperature on hydrolytic and esterifying activities from pseudomonas fragi CRDA 037 grown on whey AB - The production of hydrolytic and esterifying activities of Pseudomonas fragi CRDA 037 grown on a whey-based medium was investigated at different temperatures over time. The optimal temperature was found to be critical and different for the production of both activities. The highest hydrolytic activity was detected with bacteria cultivated at between 24 degrees C (149.2 U/liter) and 27 degrees C (133.8 U/liter), while the highest production of ethyl valerate (esterifying activity) was observed by using biomass grown at 15 degrees C (0.75 U/liter). When the fermentation temperature was increased, the incubation time necessary to reach the maximal concentration of both activities was reduced. Studies of the thermostability of both activities showed that the hydrolytic activity was more stable than the esterifying activity at 15 and 24 degrees C. Statistical analysis allowed the determination of the equations that predicted the production of hydrolytic and esterifying activities as a function of time and growth temperature. The optimal assay temperatures for the hydrolytic and esterifying activities were 37 degrees C and 12 to 15 degrees C, respectively. PMID- 10388712 TI - Identification of a new gene family expressed during the onset of sexual reproduction in the centric diatom Thalassiosira weissflogii. AB - An intriguing feature of the diatom life cycle is that sexual reproduction and the generation of genetic diversity are coupled to the control of cell size. A PCR-based cDNA subtraction technique was used to identify genes that are expressed as small cells of the centric diatom Thalassiosira weissflogii initiate gametogenesis. Ten genes that are up-regulated during the early stages of sexual reproduction have been identified thus far. Three of the sexually induced genes, Sig1, Sig2, and Sig3, were sequenced to completion and are members of a novel gene family. The three polypeptides encoded by these genes possess different molecular masses and charges but display many features in common: they share five highly conserved domains; they each contain three or more cysteine-rich epithelial growth factor (EGF)-like repeats; and they each display homology to the EGF-like region of the vertebrate extracellular matrix glycoprotein tenascin X. Interestingly, the five conserved domains appear in the same order in each polypeptide but are separated by variable numbers of nonconserved amino acids. SIG1 and SIG2 display putative regulatory domains within the nonconserved regions. A calcium-binding, EF-hand motif is found in SIG1, and an ATP/GTP binding motif is present in SIG2. The striking similarity between the SIG polypeptides and extracellular matrix components commonly involved in cell-cell interactions suggests that the SIG polypeptides may play a role in sperm-egg recognition. The SIG polypeptides are thus important molecular targets for determining when and where sexual reproduction occurs in the field. PMID- 10388713 TI - Identification and characterization of a flagellin gene from the endosymbiont of the hydrothermal vent tubeworm Riftia pachyptila. AB - The bacterial endosymbionts of the hydrothermal vent tubeworm Riftia pachyptila play a key role in providing their host with fixed carbon. Results of prior research suggest that the symbionts are selected from an environmental bacterial population, although a free-living form has been neither cultured from nor identified in the hydrothermal vent environment. To begin to assess the free living potential of the symbiont, we cloned and characterized a flagellin gene from a symbiont fosmid library. The symbiont fliC gene has a high degree of homology with other bacterial flagellin genes in the amino- and carboxy-terminal regions, while the central region was found to be nonconserved. A sequence that was homologous to that of a consensus sigma28 RNA polymerase recognition site lay upstream of the proposed translational start site. The symbiont protein was expressed in Escherichia coli, and flagella were observed by electron microscopy. A 30,000-Mr protein subunit was identified in whole-cell extracts by Western blot analysis. These results provide the first direct evidence of a motile free-living stage of a chemoautotrophic symbiont and support the hypothesis that the symbiont of R. pachyptila is acquired with each new host generation. PMID- 10388714 TI - Nisin production by a mixed-culture system consisting of Lactococcus lactis and Kluyveromyces marxianus. AB - To control the pH during antimicrobial peptide (nisin) production by a lactic acid bacterium, Lactococcus lactis subsp. lactis (ATCC11454), a novel method involving neither addition of alkali nor a separation system such as a ceramic membrane filter and electrodialyzer was developed. A mixed culture of L. lactis and Kluyveromyces marxianus, which was isolated from kefir grains, was utilized in the developed system. The interaction between lactate production by L. lactis and its assimilation by K. marxianus was used to control the pH. To utilize the interaction of these microorganisms to maintain high-level production of nisin, the kinetics of growth of, and production of lactate, acetate, and nisin by, L. lactis were investigated. The kinetics of growth of and lactic acid consumption by K. marxianus were also investigated. Because the pH of the medium could be controlled by the lactate consumption of K. marxianus and the specific lactate consumption rate of K. marxianus could be controlled by changing the dissolved oxygen (DO) concentration, a cascade pH controller coupled with DO control was developed. As a result, the pH was kept constant because the lactate level was kept low and nisin accumulated in the medium to a high level compared with that attained using other pH control strategies, such as with processes lacking pH control and those in which pH is controlled by addition of alkali. PMID- 10388716 TI - Nitrogen, carbon, and sulfur metabolism in natural thioploca samples AB - Filamentous sulfur bacteria of the genus Thioploca occur as dense mats on the continental shelf off the coast of Chile and Peru. Since little is known about their nitrogen, sulfur, and carbon metabolism, this study was undertaken to investigate their (eco)physiology. Thioploca is able to store internally high concentrations of sulfur globules and nitrate. It has been previously hypothesized that these large vacuolated bacteria can oxidize sulfide by reducing their internally stored nitrate. We examined this nitrate reduction by incubation experiments of washed Thioploca sheaths with trichomes in combination with 15N compounds and mass spectrometry and found that these Thioploca samples produce ammonium at a rate of 1 nmol min-1 mg of protein-1. Controls showed no significant activity. Sulfate was shown to be the end product of sulfide oxidation and was observed at a rate of 2 to 3 nmol min-1 mg of protein-1. The ammonium and sulfate production rates were not influenced by the addition of sulfide, suggesting that sulfide is first oxidized to elemental sulfur, and in a second independent step elemental sulfur is oxidized to sulfate. The average sulfide oxidation rate measured was 5 nmol min-1 mg of protein-1 and could be increased to 10.7 nmol min-1 mg of protein-1 after the trichomes were starved for 45 h. Incorporation of 14CO2 was at a rate of 0.4 to 0.8 nmol min-1 mg of protein 1, which is half the rate calculated from sulfide oxidation. [2-14C]acetate incorporation was 0.4 nmol min-1 mg of protein-1, which is equal to the CO2 fixation rate, and no 14CO2 production was detected. These results suggest that Thioploca species are facultative chemolithoautotrophs capable of mixotrophic growth. Microautoradiography confirmed that Thioploca cells assimilated the majority of the radiocarbon from [2-14C]acetate, with only a minor contribution by epibiontic bacteria present in the samples. PMID- 10388715 TI - Discriminant analysis of ribotype profiles of Escherichia coli for differentiating human and nonhuman sources of fecal pollution. AB - Estuarine waters receive fecal pollution from a variety of sources, including humans and wildlife. Escherichia coli is a ubiquitous bacterium in the intestines of warm-blooded animals and is used as an indicator of fecal pollution. However, its presence does not specifically differentiate sources of pollution. A total of 238 E. coli isolates from human sources (HS) and nonhuman sources (NHS) were collected from the Apalachicola National Estuarine Research Reserve, from associated sewage treatment plants, and directly from animals and tested for ribotype (RT) profile. HS and NHS isolates showed 41 and 61 RT profiles, respectively. At a similarity index of ca. 50%, HS and NHS isolates demonstrated four clusters, with the majority of HS and NHS isolates located in clusters C and D; isolates obtained directly from human and animal feces also could be grouped within these clusters. Discriminant analysis (DA) of RT profiles showed that 97% of the NHS isolates and 100% of the animal fecal isolates were correctly classified. The average rate of correct classification for HS and NHS isolates was 82%. We conclude that DA of RT profiles may be a useful method for identifying HS and NHS fecal pollution and may potentially facilitate management practices. PMID- 10388717 TI - Formation of hyodeoxycholic acid from muricholic acid and hyocholic acid by an unidentified gram-positive rod termed HDCA-1 isolated from rat intestinal microflora. AB - From the rat intestinal microflora we isolated a gram-positive rod, termed HDCA 1, that is a member of a not previously described genomic species and that is able to transform the 3alpha,6beta, 7beta-trihydroxy bile acid beta-muricholic acid into hyodeoxycholic acid (3alpha,6alpha-dihydroxy acid) by dehydroxylation of the 7beta-hydroxy group and epimerization of the 6beta-hydroxy group into a 6alpha-hydroxy group. Other bile acids that were also transformed into hyodeoxycholic acid were hyocholic acid (3alpha, 6alpha,7alpha-trihydroxy acid), alpha-muricholic acid (3alpha,6beta, 7alpha-trihydroxy acid), and omega muricholic acid (3alpha,6alpha, 7beta-trihydroxy acid). The strain HDCA-1 could not be grown unless a nonconjugated 7-hydroxylated bile acid and an unidentified growth factor produced by a Ruminococcus productus strain that was also isolated from the intestinal microflora were added to the culture medium. Germfree rats selectively associated with the strain HDCA-1 plus a bile acid-deconjugating strain and the growth factor-producing R. productus strain converted beta muricholic acid almost completely into hyodeoxycholic acid. PMID- 10388718 TI - Seasonal dynamics of bacterioplankton community structure in a eutrophic lake as determined by 5S rRNA analysis. AB - Community structure of bacterioplankton was studied during the major growth season for phytoplankton (April to October) in the epilimnion of a temperate eutrophic lake (Lake Plusssee, northern Germany) by using comparative 5S rRNA analysis. Estimates of the relative abundances of single taxonomic groups were made on the basis of the amounts of single 5S rRNA bands obtained after high resolution electrophoresis of RNA directly from the bacterioplankton. Full sequence analysis of single environmental 5S rRNAs enabled the identification of single taxonomic groups of bacteria. Comparison of partial 5S rRNA sequences allowed the detection of changes of single taxa over time. Overall, the whole bacterioplankton community showed two to eight abundant (>4% of the total 5S rRNA) taxa. A distinctive seasonal succession was observed in the taxonomic structure of this pelagic community. A rather-stable community structure, with seven to eight different taxonomic units, was observed beginning in April during the spring phytoplankton bloom. A strong reduction in this diversity occurred at the beginning of the clear-water phase (early May), when only two to four abundant taxa were observed, with one taxon dominating (up to 72% of the total 5S rRNA). The community structure during summer stagnation (June and July) was characterized by frequent changes of different dominating taxa. During late summer, a dinoflagellate bloom (Ceratium hirudinella) occurred, with Comamonas acidovorans (beta-subclass of the class Proteobacteria) becoming the dominant bacterial species (average abundance of 43% of the total 5S rRNA). Finally, the seasonal dynamics of the community structure of bacterioplankton were compared with the abundances of other major groups of the aquatic food web, such as phyto- and zooplankton, revealing that strong grazing pressure by zooplankton can reduce microbial diversity substantially in pelagic environments. PMID- 10388719 TI - Hemolysis, toxicity, and randomly amplified polymorphic DNA analysis of Stachybotrys chartarum strains. AB - Stachybotrys chartarum is an indoor air, toxigenic fungus that has been associated with a number of human and veterinary health problems. Most notable among these has been a cluster of idiopathic pulmonary hemorrhage cases that were observed in the Cleveland, Ohio, area. In this study, 16 strains of S. chartarum isolated from case (n = 8) or control (n = 8) homes in Cleveland and 12 non Cleveland strains from diverse geographic locations were analyzed for hemolytic activity, conidial toxicity, and randomly amplified polymorphic DNA banding patterns. In tests for hemolytic activity, strains were grown at 23 degrees C on wet wallboard pieces for an 8-week test period. Conidia from these wallboard pieces were subcultured on sheep's blood agar once a week over this period and examined for growth and clearing of the medium at 37 or 23 degrees C. Five of the Cleveland strains (all from case homes) showed hemolytic activity at 37 degrees C throughout the 8-week test compared to 3 of the non-Cleveland strains. Five of the Cleveland strains, compared to two of the non-Cleveland strains, produced highly toxic conidia (>90 microgram of T2 toxin equivalents per g [wet weight] of conidia) after 10 and 30 days of growth on wet wallboard. Only 3 of the 28 strains examined both were consistently hemolytic and produced highly toxic conidia. Each of these strains was isolated from a house in Cleveland where an infant had idiopathic pulmonary hemorrhage. PMID- 10388720 TI - In situ analysis of nitrifying biofilms as determined by in situ hybridization and the use of microelectrodes. AB - We investigated the in situ spatial organization of ammonia-oxidizing and nitrite oxidizing bacteria in domestic wastewater biofilms and autotrophic nitrifying biofilms by using microsensors and fluorescent in situ hybridization (FISH) performed with 16S rRNA-targeted oligonucleotide probes. The combination of these techniques made it possible to relate in situ microbial activity directly to the occurrence of nitrifying bacterial populations. In situ hybridization revealed that bacteria belonging to the genus Nitrosomonas were the numerically dominant ammonia-oxidizing bacteria in both types of biofilms. Bacteria belonging to the genus Nitrobacter were not detected; instead, Nitrospira-like bacteria were the main nitrite-oxidizing bacteria in both types of biofilms. Nitrospira-like cells formed irregularly shaped aggregates consisting of small microcolonies, which clustered around the clusters of ammonia oxidizers. Whereas most of the ammonia oxidizing bacteria were present throughout the biofilms, the nitrite-oxidizing bacteria were restricted to the active nitrite-oxidizing zones, which were in the inner parts of the biofilms. Microelectrode measurements showed that the active ammonia-oxidizing zone was located in the outer part of a biofilm, whereas the active nitrite-oxidizing zone was located just below the ammonia-oxidizing zone and overlapped the location of nitrite-oxidizing bacteria, as determined by FISH. PMID- 10388721 TI - Phylogenetic analysis of particle-attached and free-living bacterial communities in the Columbia river, its estuary, and the adjacent coastal ocean. AB - The Columbia River estuary is a dynamic system in which estuarine turbidity maxima trap and extend the residence time of particles and particle-attached bacteria over those of the water and free-living bacteria. Particle-attached bacteria dominate bacterial activity in the estuary and are an important part of the estuarine food web. PCR-amplified 16S rRNA genes from particle-attached and free-living bacteria in the Columbia River, its estuary, and the adjacent coastal ocean were cloned, and 239 partial sequences were determined. A wide diversity was observed at the species level within at least six different bacterial phyla, including most subphyla of the class Proteobacteria. In the estuary, most particle-attached bacterial clones (75%) were related to members of the genus Cytophaga or of the alpha, gamma, or delta subclass of the class Proteobacteria. These same clones, however, were rare in or absent from either the particle attached or the free-living bacterial communities of the river and the coastal ocean. In contrast, about half (48%) of the free-living estuarine bacterial clones were similar to clones from the river or the coastal ocean. These free living bacteria were related to groups of cosmopolitan freshwater bacteria (beta proteobacteria, gram-positive bacteria, and Verrucomicrobium spp.) and groups of marine organisms (gram-positive bacteria and alpha-proteobacteria [SAR11 and Rhodobacter spp.]). These results suggest that rapidly growing particle-attached bacteria develop into a uniquely adapted estuarine community and that free-living estuarine bacteria are similar to members of the river and the coastal ocean microbial communities. The high degree of diversity in the estuary is the result of the mixing of bacterial communities from the river, estuary, and coastal ocean. PMID- 10388722 TI - Phosphate stress in cultures and field populations of the dinoflagellate prorocentrum minimum detected by a single-cell alkaline phosphatase assay AB - Alkaline phosphatase activity is a common marker of phosphate stress in many phytoplankton, but it has been difficult to attribute alkaline phosphatase activity to specific organisms or groups of phytoplankton in the field with traditional biochemical procedures. A new alkaline phosphatase substrate, ELF-97 (enzyme-labeled fluorescence), shows promise in this regard. When a phosphate group is cleaved from the ELF-97 reagent, the remaining molecule precipitates near the site of enzyme activity, thus fluorescently tagging cells with alkaline phosphatase activity. We characterized ELF-97 labeling in axenic cultures of a common dinoflagellate, Prorocentrum minimum, in order to understand ELF-97 labeling dynamics when phosphate nutrition varies. Enzyme activity, as detected by ELF-97 labeling, appears to be induced in late-log- or early-stationary-phase cultures if cells are grown in low-phosphate media and is lost when phosphate stressed cells are refed with phosphate. ELF-97 appears to label an inducible intracellular alkaline phosphatase in P. minimum based on confocal microscopy studies. This may limit the use of this reagent to organisms that lack high levels of constitutive intracellular phosphatases. After laboratory cultures were characterized, ELF-97 was used to assay field populations of P. minimum in Narragansett Bay during two 1-week periods, and 12 to 100% of the P. minimum cells were labeled. The level of cell labeling was reduced by 3 days of incubation with added inorganic phosphate. Our results indicate that ELF-97 is an excellent new tool for monitoring phytoplankton phosphate stress in the environment when the data are supported by appropriate laboratory studies. PMID- 10388723 TI - Use of conserved randomly amplified polymorphic DNA (RAPD) fragments and RAPD pattern for characterization of Lactobacillus fermentum in Ghanaian fermented maize dough. AB - The present work describes the use of randomly amplified polymorphic DNA (RAPD) for the characterization of 172 dominant Lactobacillus isolates from present and previous studies of Ghanaian maize fermentation. Heterofermentative lactobacilli dominate the fermentation flora, since approximately 85% of the isolates belong to this group. Cluster analysis of the RAPD profiles obtained showed the presence of two main clusters. Cluster 1 included Lactobacillus fermentum, whereas cluster 2 comprised the remaining Lactobacillus spp. The two distinct clusters emerged at the similarity level of <50%. All isolates in cluster 1 showed similarity in their RAPD profile to the reference strains of L. fermentum included in the study. These isolates, yielding two distinct bands of approximately 695 and 773 bp with the primers used, were divided into four subclusters, indicating that several strains are involved in the fermentation and remain dominant throughout the process. The two distinct RAPD fragments were cloned, sequenced, and used as probes in Southern hybridization experiments. With one exception, Lactobacillus reuteri LMG 13045, the probes hybridized only to fragments of different sizes in EcoRI-digested chromosomal DNA of L. fermentum strains, thus indicating the specificity of the probes and variation within the L. fermentum isolates. PMID- 10388724 TI - Electrophoretic mobilities of Escherichia coli O157:H7 and wild-type Escherichia coli strains. AB - The electrophoretic mobilities (EPMs) of a number of Escherichia coli O157:H7 and wild-type E. coli strains were measured. The effects of pH and ionic strength on the EPMs were investigated. The EPMs of E. coli O157:H7 strains differed from those of wild-type strains. As the suspension pH decreased, the EPMs of both types of strains increased. PMID- 10388725 TI - Specific detection of the gene for the extracellular neutral protease of Bacillus cereus by PCR and blot hybridization. AB - A pair of primers and a gene probe for the amplification and detection of the Bacillus cereus neutral protease gene (NPRC) were developed. Specificity for the npr genes of the B. cereus group members B. cereus, B. mycoides, and B. thuringiensis was shown. Restriction polymorphism patterns of the PCR products confirmed the presence of the NPRC gene in all three species. PMID- 10388726 TI - Viability and virulence of experimentally stressed nonculturable Salmonella typhimurium. AB - Maintenance of pathogenicity of viable but nonculturable Salmonella typhimurium cells experimentally stressed with UV-C and seawater, was investigated relative to the viability level of the cellular population. Pathogenicity, tested in a mouse model, was lost concomitantly with culturability, whereas cell viability remained undamaged, as determined by respiratory activity and cytoplasmic membrane and genomic integrities. PMID- 10388727 TI - Effect of cattle diet on Escherichia coli O157:H7 acid resistance. AB - The duration of shedding of Escherichia coli O157 isolates by hay-fed and grain fed steers experimentally inoculated with E. coli O157:H7 was compared, as well as the acid resistance of the bacteria. The hay-fed animals shed E. coli O157 longer than the grain-fed animals, and irrespective of diet, these bacteria were equally acid resistant. Feeding cattle hay may increase human infections with E. coli O157:H7. PMID- 10388728 TI - Comparison of sensitivity of immunofluorescent microscopy to that of a combination of immunofluorescent microscopy and immunomagnetic separation for detection of Cryptosporidium parvum oocysts in adult bovine feces. AB - A direct immunofluorescence assay (DFA) (Merifluor; Meridian Diagnostics, Inc., Cincinnati, Ohio) was compared to an immunomagnetic separation (IMS) assay (Dynabeads; Dynal, Inc., Lake Success, N.Y.) coupled with immunofluorescent microscopy (Waterborne, Inc., New Orleans, La.) for their ability to detect low concentrations of Cryptosporidium parvum oocysts in adult bovine fecal material. IMS-DFA resulted in a 2-log-unit increase in sensitivity (10 oocysts/g) compared to DFA alone (1,000 oocysts/g). The higher sensitivity obtained with IMS-DFA resulted from testing 2 g of fecal material instead of the 13 to 19 mg of fecal material tested in the DFA; the increased sensitivity was not attributable to a higher percent recovery. PMID- 10388729 TI - In situ detection of the Clostridium botulinum type C1 toxin gene in wetland sediments with a nested PCR assay. AB - A nested PCR was developed for detection of the Clostridium botulinum type C1 toxin gene in sediments collected from wetlands where avian botulism outbreaks had or had not occurred. The C1 toxin gene was detected in 16 of 18 sites, demonstrating both the ubiquitous distribution of C. botulinum type C in wetland sediments and the sensitivity of the detection assay. PMID- 10388730 TI - Characterization of methylglyoxal synthase from Clostridium acetobutylicum ATCC 824 and its use in the formation of 1, 2-propanediol. AB - A gene encoding a putative 150-amino-acid methylglyoxal synthase was identified in Clostridium acetobutylicum ATCC 824. The enzyme was overexpressed in Escherichia coli and purified. Methylglyoxal synthase has a native molecular mass of 60 kDa and an optimum pH of 7.5. The Km and Vmax values for the substrate dihydroxyacetone phosphate were 0.53 mM and 1.56 mmol min(-1) microgram(-1), respectively. When E. coli glycerol dehydrogenase was coexpressed with methylglyoxal synthase in E. coli BL21(DE3), 3.9 mM 1,2-propanediol was produced. PMID- 10388731 TI - Key physiology of anaerobic ammonium oxidation. AB - The physiology of anaerobic ammonium oxidizing (anammox) aggregates grown in a sequencing batch reactor was investigated quantitatively. The physiological pH and temperature ranges were 6.7 to 8.3 and 20 to 43 degrees C, respectively. The affinity constants for the substrates ammonium and nitrite were each less than 0.1 mg of nitrogen per liter. The anammox process was completely inhibited by nitrite concentrations higher than 0.1 g of nitrogen per liter. Addition of trace amounts of either of the anammox intermediates (1. 4 mg of nitrogen per liter of hydrazine or 0.7 mg of nitrogen per liter of hydroxylamine) restored activity completely. PMID- 10388732 TI - Determination of total protein content of bacterial cells by SYPRO staining and flow cytometry. AB - An assay has been developed for measuring protein biomass of marine planktonic bacteria by flow cytometry. The method was calibrated by using five species of Bacteria (an Arcobacter sp., a Cytophaga sp., an Oceanospirillum sp., a Pseudoalteromonas sp., and a Vibrio sp.) recently isolated from seawater samples and grown in culture at different temperatures. The intensity of SYPRO-protein fluorescence of these bacteria strongly correlated with their total protein content, measured by the bicinchoninic acid method to be in the range of 60 to 330 fg of protein cell-1 (r2 = 0.93, n = 34). According to the calibration, the mean biomass of planktonic bacteria from the North Sea in August 1998 was 24 fg of protein cell-1. PMID- 10388733 TI - Influence of 1-[(E)-2-(2-methyl-4-nitrophenyl)diaz-1-enyl]pyrrolidine-2 carboxylic acid and diphenyliodonium chloride on ruminal protein metabolism and ruminal microorganisms. AB - The effects of 1-[(E)-2-(2-methyl-4-nitrophenyl)diaz-1-enyl]pyrrolidine-2-carboxy lic acid (LY29) and diphenyliodonium chloride (DIC) on the degradation of protein to ammonia were determined in a mixed rumen microbial population taken from sheep on a grass hay-concentrate diet. Both compounds decreased NH3 production by inhibiting deamination of amino acids. LY29, but not DIC, inhibited growth of the high-activity ammonia-producing species, Clostridium aminophilum and Clostridium sticklandii. PMID- 10388734 TI - Development of bacterial contamination during production of yeast extracts. AB - Baker's yeast suspensions having bacterial populations of 10(6) and 10(8) CFU/ml were subjected to autolysis processes designed to obtain yeast extracts (YE). The bacterial contaminants added to the yeast cell suspensions were produced with spent broths obtained from a commercial yeast production plant and contained 59% cocci (Leuconostoc, Aerococcus, Lactococcus) as well as 41% bacilli (Bacillus). Autolyses were conducted at four different pH levels (4.0, 5.5, 7.0, and 8.5) and with two autolysis-promoting agents (ethyl acetate and chitosan). Processing parameters were more important than the initial bacterial population in the development of contaminating bacteria during manufacture of YE. Drops in the viable bacterial population after a 24-h autolysis were observed when pH was adjusted to 4.0 or when ethyl acetate was added. A significant interaction was found between the effects of pH and autolysis promoters on the bacterial population in YE, indicating that the activity of ethyl acetate, as opposed to that of chitosan, was not influenced by pH. PMID- 10388735 TI - The iso-competition point, a new concept for characterizing multivalent versus monovalent counterion competition, successfully describes cation binding to DNA. PMID- 10388736 TI - A self-consistent, microenvironment modulated screened coulomb potential approximation to calculate pH-dependent electrostatic effects in proteins. AB - An improved approach is presented for calculating pH-dependent electrostatic effects in proteins using sigmoidally screened Coulomb potentials (SCP). It is hypothesized that a key determinant of seemingly aberrant behavior in pKa shifts is due to the properties of the unique microenvironment around each residue. To help demonstrate this proposal, an approach is developed to characterize the microenvironments using the local hydrophobicity/hydrophilicity around each residue of the protein. The quantitative characterization of the microenvironments shows that the protein is a complex mosaic of differing dielectric regions that provides a physical basis for modifying the dielectric screening functions: in more hydrophobic microenvironments the screening decreases whereas the converse applies to more hydrophilic regions. The approach was applied to seven proteins providing more than 100 measured pKa values and yielded a root mean square deviation of 0.5 between calculated and experimental values. The incorporation of the local hydrophobicity characteristics into the algorithm allowed the resolution of some of the more intractable problems in the calculation of pKa. Thus, the divergent shifts of the pKa of Glu-35 and Asp-66 in hen egg white lysozyme, which are both about 90% buried, was correctly predicted. Mechanistically, the divergence occurs because Glu-35 is in a hydrophobic microenvironment, while Asp-66 is in a hydrophilic microenvironment. Furthermore, because the calculation of the microenvironmental effects takes very little CPU time, the computational speed of the SCP formulation is conserved. Finally, results from different crystal structures of a given protein were compared, and it is shown that the reliability of the calculated pKa values is sufficient to allow identification of conformations that may be more relevant for the solution structure. PMID- 10388737 TI - Generalization of the theory of transition times in metabolic pathways: a geometrical approach. AB - Cell metabolism is able to respond to changes in both internal parameters and boundary constraints. The time any system variable takes to make this response has relevant implications for understanding the evolutionary optimization of metabolism as well as for biotechnological applications. This work is focused on estimating the magnitude of the average time taken by any observable of the system to reach a new state when either a perturbation or a persistent variation occurs. With this aim, a new variable, called characteristic time, based on geometric considerations, is introduced. It is stressed that this new definition is completely general, being useful for evaluating the response time, even in complex transitions involving periodic behavior. It is shown that, in some particular situations, this magnitude coincides with previously defined transition times but differs drastically in others. Finally, to illustrate the applicability of this approach, a model of a reaction mediated by an allosteric enzyme is analyzed. PMID- 10388738 TI - Impact of mitochondrial Ca2+ cycling on pattern formation and stability. AB - Energization of mitochondria significantly alters the pattern of Ca2+ wave activity mediated by activation of the inositol (1,4,5) trisphosphate (IP3) receptor (IP3R) in Xenopus oocytes. The number of pulsatile foci is reduced and spiral Ca2+ waves are no longer observed. Rather, target patterns of Ca2+ release predominate, and when fragmented, fail to form spirals. Ca2+ wave velocity, amplitude, decay time, and periodicity are also increased. We have simulated these experimental findings by supplementing an existing mathematical model with a differential equation for mitochondrial Ca2+ uptake and release. Our calculations show that mitochondrial Ca2+ efflux plays a critical role in pattern formation by prolonging the recovery time of IP3Rs from a refractory state. We also show that under conditions of high energization of mitochondria, the Ca2+ dynamics can become bistable with a second stable stationary state of high resting Ca2+ concentration. PMID- 10388739 TI - Effect of Na/Ca exchange on plateau fraction and [Ca]i in models for bursting in pancreatic beta-cells. AB - In the presence of an insulinotropic glucose concentration, beta-cells, in intact pancreatic islets, exhibit periodic bursting electrical activity consisting of an alternation of active and silent phases. The fraction of time spent in the active phase over a period is called the plateau fraction and is correlated with the rate of insulin release. However, the mechanisms that regulate the plateau fraction remain unclear. In this paper we investigate the possible role of the plasma membrane Na+/Ca2+ exchange of the beta-cell in controlling the plateau fraction. We have extended different single-cell models to incorporate this Ca2+ activated electrogenic Ca2+ transporter. We find that the Na+/Ca2+ exchange can provide a physiological mechanism to increase the plateau fraction as the glucose concentration is raised. In addition, we show theoretically that the Na+/Ca2+ exchanger is a key regulator of the cytoplasmic calcium concentration in clusters of heterogeneous cells with gap-junctional electrical coupling. PMID- 10388740 TI - De novo simulations of the folding thermodynamics of the GCN4 leucine zipper. AB - Entropy Sampling Monte Carlo (ESMC) simulations were carried out to study the thermodynamics of the folding transition in the GCN4 leucine zipper (GCN4-lz) in the context of a reduced model. Using the calculated partition functions for the monomer and dimer, and taking into account the equilibrium between the monomer and dimer, the average helix content of the GCN4-lz was computed over a range of temperatures and chain concentrations. The predicted helix contents for the native and denatured states of GCN4-lz agree with the experimental values. Similar to experimental results, our helix content versus temperature curves show a small linear decline in helix content with an increase in temperature in the native region. This is followed by a sharp transition to the denatured state. van't Hoff analysis of the helix content versus temperature curves indicates that the folding transition can be described using a two-state model. This indicates that knowledge-based potentials can be used to describe the properties of the folded and unfolded states of proteins. PMID- 10388741 TI - A study of vibrational relaxation of B-state carbon monoxide in the heme pocket of photolyzed carboxymyoglobin. AB - The vibrational energy relaxation of dissociated carbon monoxide in the heme pocket of sperm whale myoglobin has been studied using equilibrium molecular dynamics simulation and normal mode analysis methods. Molecular dynamics trajectories of solvated myoglobin were run at 300 K for both the delta- and epsilon-tautomers of the distal histidine, His64. Vibrational population relaxation times were estimated using the Landau-Teller model. For carbon monoxide (CO) in the myoglobin epsilon-tautomer, for a frequency of omega0 = 2131 cm-1 corresponding to the B1 state, T1epsilon(B1) = 640 +/- 185 ps, and for a frequency of omega0 = 2119 cm-1 corresponding to the B2 state, T1epsilon(B2) = 590 +/- 175 ps. Although the CO relaxation rates in both the epsilon- and delta tautomers are similar in magnitude, the simulations predict that the vibrational relaxation of the CO is faster in the delta-tautomer. For CO in the myoglobin delta-tautomer, it was found that the relaxation times were identical within error for the two CO substate frequencies, T1delta(B1) = 335 +/- 115 ps and T1delta(B2) = 330 +/- 145 ps. These simulation results are in reasonable agreement with experimental results of Anfinrud and coworkers (unpublished results). Normal mode calculations were used to identify the dominant coupling between the protein and CO molecules. The calculations suggest that the residues of the myoglobin pocket, acting as a first solvation shell to the CO molecule, contribute the primary "doorway" modes in the vibrational relaxation of the oscillator. PMID- 10388742 TI - Consequences of breaking the Asp-His hydrogen bond of the catalytic triad: effects on the structure and dynamics of the serine esterase cutinase. AB - The objective of this study has been to investigate the effects on the structure and dynamics that take place with the breaking of the Asp-His hydrogen bond in the catalytic triad Asp175-His188-Ser120 of the serine esterase cutinase in the ground state. Four molecular dynamics simulations were performed on this enzyme in solution. The starting structures in two simulations had the Asp175-His188 hydrogen bond intact, and in two simulations the Asp175-His188 hydrogen bond was broken. Conformations of the residues comprising the catalytic triad are well behaved during both simulations containing the intact Asp175-His188 hydrogen bond. Short contacts of less than 2.6 A were observed in 1.2% of the sampled distances between the carboxylate oxygens of Asp175 and the NE2 of His188. The simulations showed that the active site residues exhibit a great deal of mobility when the Asp175-His188 hydrogen bond is broken. In the two simulations in which the Asp175-His188 hydrogen bond is not present, the final geometries for the residues in the catalytic triad are not in catalytically productive conformations. In both simulations, Asp175 and His188 are more than 6 A apart in the final structure from dynamics, and the side chains of Ser120 and Asp175 are in closer proximity to the NE2 of His188 than to ND1. Nonlocal effects on the structure of cutinase were observed. A loop formed by residues 26-31, which is on the opposite end of the protein relative to the active site, was greatly affected. Further changes in the dynamics of cutinase were determined from quasiharmonic mode analysis. The frequency of the second lowest mode was greatly reduced when the Asp175-His188 hydrogen bond was broken, and several higher modes showed lower frequencies. All four simulations showed that the oxyanion hole, composed of residues Ser42 and Gln121, is stable. Only one of the hydrogen bonds (Ser42 OG to Gln121 NE2) observed in the crystal structure that stabilize the conformation of Ser42 OG persisted throughout the simulations. This hydrogen bond appears to be enough for the oxyanion hole to retain its structural integrity. PMID- 10388743 TI - Probing the structural changes in the light chain of human coagulation factor VIIa due to tissue factor association. AB - The crystallographic structure of human coagulation factor VIIa/tissue factor complex bound with calcium ions was used to model the solution structure of the light chain of factor VIIa (residues 1-142) in the absence of tissue factor. The Amber force field in conjunction with the particle mesh Ewald summation method to accommodate long-range electrostatic interactions was used in the trajectory calculations. The estimated TF-free solution structure was then compared with the crystal structure of factor VIIa/tissue factor complex to estimate the restructuring of factor VIIa due to tissue factor binding. The solution structure of the light chain of factor VIIa in the absence of tissue factor is predicted to be an extended domain structure similar to that of the tissue factor-bound crystal. Removal of the EGF1-bound calcium ion is shown by simulation to lead to minor structural changes within the EGF1 domain, but also leads to substantial relative reorientation of the Gla and EGF1 domains. PMID- 10388744 TI - The iso-competition point for counterion competition binding to DNA: calculated multivalent versus monovalent cation binding equivalence. AB - In this paper we introduce an important parameter called the iso-competition point (ICP), to characterize the competition binding to DNA in a two-cation species system. By imposing the condition of charge neutralization fraction equivalence theta1 = ZthetaZ upon the two simultaneous equations in Manning's counterion condensation theory, the ICPs can be calculated. Each ICP, which refers to a particular multivalent concentration where the charge fraction on DNA neutralized from monovalent cations equals that from the multivalent cations, corresponds to a specific ionic strength condition. At fixed ionic strength, the total DNA charge neutralization fractions thetaICP are equal, no matter whether the higher valence cation is divalent, trivalent, or tetravalent. The ionic strength effect on ICP can be expressed by a semiquantitative equation as ICPZa/ICPZb = (Ia/Ib)Z, where Ia, Ib refers to the instance of ionic strengths and Z indicates the valence. The ICP can be used to interpret and characterize the ionic strength, valence, and DNA length effects on the counterion competition binding in a two-species system. Data from our previous investigations involving binding of Mg2+, Ca2+, and Co(NH3)63+ to lambda-DNA-HindIII fragments ranging from 2.0 to 23.1 kbp was used to investigate the applicability of ICP to describe counterion binding. It will be shown that the ICP parameter presents a prospective picture of the counterion competition binding to polyelectrolyte DNA under a specific ion environment condition. PMID- 10388745 TI - Change in conformation by DNA-peptide association: molecular dynamics of the Hin recombinase-hixL complex. AB - The Hin-DNA complex is a molecular complex formed by the C-terminal 52mer peptide of the Hin-recombinase and a synthetic 13-bp hixL DNA. The peptide has three alpha-helices, the second and third of which form the helix-turn-helix motif to bind to the major groove. Both termini of the peptide reside within the minor groove. Three molecular dynamics simulations were performed based on the crystal structure of the Hin-DNA complex: one for the free Hin peptide, one for the free hixL DNA, and one for the complex. Analyses of the trajectories revealed that the dynamic fluctuations of both the Hin peptide and the hixL DNA were lowered by the complex formation. The simulation supported the experimental observation that the N-terminus and the helix-turn-helix motif were critical for formation of the complex, but the C-terminus played only a supportive role in DNA recognition. The simulations strongly suggested that the binding reaction should proceed by the induced fit mechanism. The ion and solvent distributions around the molecules were also examined. PMID- 10388746 TI - Statistical mechanical equilibrium theory of selective ion channels. AB - A rigorous statistical mechanical formulation of the equilibrium properties of selective ion channels is developed, incorporating the influence of the membrane potential, multiple occupancy, and saturation effects. The theory provides a framework for discussing familiar quantities and concepts in the context of detailed microscopic models. Statistical mechanical expressions for the free energy profile along the channel axis, the cross-sectional area of the pore, and probability of occupancy are given and discussed. In particular, the influence of the membrane voltage, the significance of the electric distance, and traditional assumptions concerning the linearity of the membrane electric field along the channel axis are examined. Important findings are: 1) the equilibrium probabilities of occupancy of multiply occupied channels have the familiar algebraic form of saturation properties which is obtained from kinetic models with discrete states of denumerable ion occupancy (although this does not prove the existence of specific binding sites; 2) the total free energy profile of an ion along the channel axis can be separated into an intrinsic ion-pore free energy potential of mean force, independent of the transmembrane potential, and other contributions that arise from the interfacial polarization; 3) the transmembrane potential calculated numerically for a detailed atomic configuration of the gramicidin A channel embedded in a bilayer membrane with explicit lipid molecules is shown to be closely linear over a distance of 25 A along the channel axis. Therefore, the present analysis provides some support for the constant membrane potential field approximation, a concept that has played a central role in the interpretation of flux data based on traditional models of ion permeation. It is hoped that this formulation will provide a sound physical basis for developing nonequilibrium theories of ion transport in selective biological channels. PMID- 10388747 TI - Kinetic analysis of high affinity forms of interleukin (IL)-13 receptors: suppression of IL-13 binding by IL-2 receptor gamma chain. AB - Interleukin-13 (IL-13) is a pleiotropic cytokine that controls growth, differentiation, and apoptosis of immune and tumor cells. To understand the mechanisms of interaction between IL-13 and IL-13 receptors (IL-13R), and the role of the IL-2 receptor common gamma chain (gammac) in IL-13 binding and processing, we have examined IL-13 binding kinetics, dissociation/shedding, and internalization in renal cell carcinoma (RCC) cell lines. We observed a new phenomena in that the apparent rate of association, but not the dissociation, was strongly related to IL-13 concentration. We also observed cooperativity phenomena in IL-13 and IL-13R interaction in control RCC (MLneo) cells, but not in cells transfected with gammac chain (MLgammac). The number of IL-13 binding sites, the effective rate of ligand association, and the dissociation rate constants were reduced in gammac-transfected cells compared to control RCC cells. Two forms of IL-13R were detected in these cell lines, which differed in the kinetics of endocytosis and dissociation/exocytosis. Only a small fraction of bound receptors (14-24%) was rapidly internalized and the same fraction of the ligand-receptor complexes was shed and/or dissociated. The expression of gammac chain did not change any of these processes. A two independent high-affinity and moderate affinity receptor model fit the kinetic observations in gammac-transfected cells. However, in control cells, the binding kinetics were more complicated. A mathematical model that fit a set of kinetic and steady state data in control cells was selected from a set of possible models. This best-fit model predicts that 1) two different IL-13R are expressed on the cell membrane, 2) a minor fraction of IL-13R exist as microclusters (homodimers and/or heterodimers) without exogenous IL-13, 3) high morphological complexity of the gammac-negative control cell membrane affects the cooperativity phenomena of IL-13 binding, and 4) a large number of co-receptor molecules is present, which helps keep the ligand on the cell surface for a long period of time after fast IL-13 binding and provides a negative control for ligand binding via production of the high affinity inhibitor bound to IL-13. Our data demonstrate that gammac exerts dramatic changes in the kinetic mechanisms of IL-13 binding. PMID- 10388748 TI - Kinetic mechanisms of inhibitor binding: relevance to the fast-acting slow binding paradigm. AB - Although phlorizin inhibition of Na+-glucose cotransport occurs within a few seconds, 3H-phlorizin binding to the sodium-coupled glucose transport protein(s) requires several minutes to reach equilibrium (the fast-acting slow-binding paradigm). Using kinetic models of arbitrary dimension that can be reduced to a two-state diagram according to Cha's formalism, we show that three basic mechanisms of inhibitor binding can be identified whereby the inhibitor binding step either (A) represents, (B) precedes, or (C) follows the rate-limiting step in a binding reaction. We demonstrate that each of mechanisms A-C is associated with a set of unique kinetic properties, and that the time scale over which one may expect to observe mechanism C is conditioned by the turnover number of the catalytic cycle. In contrast, mechanisms A and B may be relevant to either fast acting or slow-binding inhibitors. However, slow-binding inhibition according to mechanism A may not be compatible with a fast-acting behavior on the steady-state time scale of a few seconds. We conclude that the recruitment hypothesis (mechanism C) cannot account for slow phlorizin binding to the sodium-coupled glucose transport protein(s), and that mechanism B is the only alternative that may explain the fast-acting slow-binding paradigm. PMID- 10388749 TI - Activation and inhibition of skeletal RyR channels by a part of the skeletal DHPR II-III loop: effects of DHPR Ser687 and FKBP12. AB - Peptides, corresponding to sequences in the N-terminal region of the skeletal muscle dihydropyridine receptor (DHPR) II-III loop, have been tested on sarcoplasmic reticulum (SR) Ca2+ release and ryanodine receptor (RyR) activity. The peptides were: A1, Thr671-Leu690; A2, Thr671-Leu690 with Ser687 Ala substitution; NB, Gly689-Lys708 and A1S, scrambled A1 sequence. The relative rates of peptide-induced Ca2+ release from normal (FKBP12+) SR were A2 > A1 > A1S > NB. Removal of FKBP12 reduced the rate of A1-induced Ca2+ release by approximately 30%. A1 and A2 (but not NB or A1S), in the cytoplasmic (cis) solution, either activated or inhibited single FKBP12+ RyRs. Maximum activation was seen at -40 mV, with 10 microM A1 or 50 nM A2. The greatest A1-induced increase in mean current (sixfold) was seen with 100 nM cis Ca2+. Inhibition by A1 was greatest at +40 mV (or when permeant ions flowed from cytoplasm to SR lumen) with 100 microM cis Ca2+, where channel activity was almost fully inhibited. A1 did not activate FKBP12-stripped RyRs, although peptide-induced inhibition remained. The results show that peptide A activation of RyRs does not require DHPR Ser687, but required FKBP12 binding to RyRs. Peptide A must interact with different sites to activate or inhibit RyRs, because current direction-, voltage-, cis [Ca2+]-, and FKBP12-dependence of activation and inhibition differ. PMID- 10388750 TI - Effect of protein kinase A-induced phosphorylation on the gating mechanism of the brain Na+ channel: model fitting to whole-cell current traces. AB - The activity of the voltage-gated Na+ channel is subjected to modulation through covalent modifications. It has been previously shown that brain Na+ currents are reduced following the activation of the protein kinase A (PKA) pathway, but the effect of the phosphorylation on the gating mechanism of the channel has not been demonstrated so far. In this study, we analyze the whole-cell Na+ current recorded in the absence or presence of forskolin, which stimulates the PKA pathway. A minimal molecular model of the gating mechanism of the Na+ channel is defined to fit the experimental data: it consists of three closed states, one open state, and two inactivated states. We experimentally demonstrate that the kinetics of inactivation from the closed states are not affected by phosphorylation. The results obtained by computer fitting indicate that, among all the kinetic parameters describing the transitions between states, only one parameter is significantly modified in the presence of forskolin, and corresponds to the acceleration of the inactivation from the open state. This conclusion is supported by the analysis of current traces obtained from cells in the presence of a phosphatase inhibitor or loaded with the PKA catalytic unit, and is in agreement with previously reported single channel records. PMID- 10388751 TI - Permeability of single nuclear pores. AB - In this first application of optical single transporter recording (OSTR), a recently established technique for optically monitoring the activity of single transporters in membrane patches (Tschodrich-Rotter and Peters. 1998. J. Microsc. 192:114-125), the passive permeability of the nuclear pore complex (NPC) was measured for a homologous series of hydrophilic probe molecules. Nuclei were isolated from Xenopus oocytes and firmly attached to filters containing small cylindrical pores. Transport through membrane patches spanning filter pores was measured by scanning microphotolysis. Thus the permeability coefficients of single NPCs were determined for fluorescently labeled dextrans of approximately 4, 10, and 20 kDa. Dextrans of >/=40 kDa could not permeate the NPC. The data were consistent with a model in which the NPC contains a single diffusion channel. By application of established theories for the restricted diffusion through small pores, the diffusion channel was approximated as a cylinder with a radius of 4.4-6.1 nm (mean 5. 35 nm). Because the transport rate constant of the single NPC was known, the equivalent length of the channel could be also determined and was found to be 40-50 nm (mean 44.5 nm). The symmetry of the NPC implies that a singular component such as the diffusion channel is located at the center of the NPC. Therefore a common transport pathway apparently mediates both passive and signal-dependent transport. To test this hypothesis, measurements of signal-dependent transport and of the mutual effects signal-dependent and passive transport may exert on each other are in progress. PMID- 10388752 TI - Tonic and phasic tetrodotoxin block of sodium channels with point mutations in the outer pore region. AB - Tonic and use-dependent block by tetrodotoxin (TTX) has been studied in cRNA injected Xenopus oocytes expressing mutants W386Y, E945Q, D1426K, and D1717Q, of the outer-pore region of the rat brain IIA alpha-subunit of sodium channels. The various phenotypes are tonically half-blocked at TTX concentrations, IC50(t), that span a range of more than three orders of magnitude, from 4 nM in mutant D1426K to 11 microM in mutant D1717Q. When stimulated with repetitive depolarizing pulses at saturating frequencies, all channels showed a monoexponential increase in their TTX-binding affinity with time constants that span an equally wide range of values ([TTX] approximately IC50(t), from approximately 60 s for D1426K to approximately 30 ms for D1717Q) and are in most phenotypes roughly inversely proportional to IC50(t). In contrast, all phenotypes show the same approximately threefold increase in their TTX affinity under stimulation. The invariance of the free-energy difference between tonic and phasic configurations of the toxin-receptor complex, together with the extreme variability of phasic block kinetics, is fully consistent with the trapped-ion mechanism of use dependence suggested by and developed by. Using this model, we estimated for each phenotype both the second-order association rate constant, kon, and the first-order dissociation rate constant, koff, for TTX binding. Except for mutant E945Q, all phenotypes have roughly the same value of kon approximately 2 microM-1 s-1 and owe their large differences in IC50(t) to different koff values. However, a 60-fold reduction in kon is the main determinant of the low TTX sensitivity of mutant E945Q. This suggests that the carboxyl group of E945 occupies a much more external position in the pore vestibule than that of the homologous residue D1717. PMID- 10388753 TI - Conformational changes of the in situ nuclear pore complex. AB - By bridging the double membrane separating the cell nucleus and cytoplasm, nuclear pore complexes (NPCs) are crucial pathways for the exchange of ions, proteins, and RNA between these two cellular compartments. A structure in the central lumen of the NPC, called the nuclear transport protein, central granule, or nuclear plug, appeared to gate diffusion of intermediate-sized molecules (10 40 kDa) across the nuclear membranes. Visualization of the NPC required drying and fixation of the specimen for electron and atomic force microscopy (AFM), a requirement that has raised doubts about the physiological relevance of the observation. Here we present AFM images of the outer nuclear membranes and NPCs of Xenopus laevis oocytes under more physiological conditions. Measured under a variety of Ca2+ depletion conditions, the central granule appeared to occupy and occlude the lumen of the pore in >80% of NPCs compared to <10% in controls. In a few instances images were obtained of the same NPCs as the solution was changed from control saline to store depletion conditions, and finally to store repletion conditions. We conclude that the central lumen of the nuclear pore complex undergoes a conformational change in response to depletion of nuclear cisternal Ca2+ levels. PMID- 10388754 TI - Heteromeric assembly of Kv2.1 with Kv9.3: effect on the state dependence of inactivation. AB - Modulatory alpha-subunits of Kv channels remain electrically silent after homomeric expression. Their interactions with Kv2 alpha-subunits via the amino terminal domain promote the assembly of heteromeric functional channels. The kinetic features of these heteromers differ from those of Kv2 homomers, suggesting a distinct role in electrical signaling. This study investigates biophysical properties of channels emerging from the coexpression of Kv2.1 with the modulatory alpha-subunit Kv9.3. Changes relative to homomeric Kv2.1 concern activation, deactivation, inactivation, and recovery from inactivation. A detailed description of Kv2.1/Kv9.3 inactivation is presented. Kv2.1/Kv9.3 heteromers inactivate in a fast and complete fashion from intermediate closed states, but in a slow and incomplete manner from open states. Intermediate closed states of channel gating can be approached through partial activation or deactivation, according to a proposed qualitative model. These transitions are rate-limiting for Kv2.1/Kv9.3 inactivation. Finally, based on the kinetic description, we propose a putative function for Kv2.1/Kv9.3 heteromers in rat heart. PMID- 10388755 TI - Catalytic activity of an isolated domain of Na,K-ATPase expressed in Escherichia coli. AB - Fusion proteins of glutathione-S-transferase and fragments from the large cytoplasmic domain of the sheep Na,K-ATPase alpha1-subunit were expressed in Escherichia coli. The Na,K-ATPase sequences begin at Ala345 and terminate at either Arg600 (DP600f), Thr610 (DP610f), Gly731 (DP731f), or Glu779 (DP779f). After affinity purification on glutathione-Sepharose, the fusion proteins were labeled with [alpha-32P]-2-N3-ATP, and incorporation of the radiolabel into the fusion proteins was measured by scintillation counting after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Kd values of 220-290 microM for 2-N3 ATP binding to the fusion proteins were obtained from the photolabeling experiments. Approximately 1 mol of 2-N3-ATP was calculated to be incorporated per mole of fusion protein after correction for photochemical incorporation efficiency. Labeling of all of the fusion proteins by 25 microM 2-N3-ATP was reduced in the presence of MgATP, Na2ATP, MgCl2, 2',3'-O-(2,4, 6-trinitrophenyl) ATP, and p-nitrophenylphosphate, and Ki values of 2-11 mM for Na2ATP, 0.2-5 mM for MgCl2, 0.1-5 mM for MgATP, and 20-300 microM for p-nitrophenylphosphate were calculated for these ligands. All of the fusion proteins catalyze the hydrolysis of p-nitrophenylphosphate. The reaction requires MgCl2 and is inhibited by inorganic phosphate, which is similar to the hydrolysis of p-nitrophenylphosphate by native Na,K-ATPase. Based on these observations, it appears that the soluble fragments from the large cytoplasmic domain of Na,K-ATPase expressed in bacterial cells are folded in an E2-like conformation and are likely to retain much of the native structure. PMID- 10388756 TI - Hofmeister effects of anions on the kinetics of partial reactions of the Na+,K+ ATPase. AB - The effects of lyotropic anions, particularly perchlorate, on the kinetics of partial reactions of the Na+,K+-ATPase from pig kidney were investigated by two different kinetic techniques: stopped flow in combination with the fluorescent label RH421 and a stationary electrical relaxation technique. It was found that 130 mM NaClO4 caused an increase in the Kd values of both the high- and low affinity ATP-binding sites, from values of 7.0 (+/- 0.6) microM and 143 (+/- 17) microM in 130 mM NaCl solution to values of 42 (+/- 3) microM and 660 (+/- 100) microM in 130 mM NaClO4 (pH 7.4, 24 degrees C). The half-saturating concentration of the Na+-binding sites on the E1 conformation was found to decrease from 8-10 mM in NaCl to 2.5-3.5 mM in NaClO4 solution. The rate of equilibration of the reaction, E1P(Na+)3 left arrow over right arrow E2P + 3Na+, decreased from 393 (+/- 51) s-1 in NaCl solution to 114 (+/- 15) s-1 in NaClO4. This decrease is attributed predominantly to an inhibition of the E1P(Na+)3 --> E2P(Na+)3 transition. The effects can be explained in terms of electrostatic interactions due to perchlorate binding within the membrane and/or protein matrix of the Na+,K+-ATPase membrane fragments and alteration of the local electric field strength experienced by the protein. The kinetic results obtained support the conclusion that the conformational transition E1P(Na+)3 --> E2P(Na+)3 is a major charge translocating step of the pump cycle. PMID- 10388757 TI - Proton and zinc effects on HERG currents. AB - The proton and Zn2+ effects on the human ether-a-go-go related gene (HERG) channels were studied after expression in Xenopus oocytes and stable transfection in the mammalian L929 cell line. Experiments were carried out using the two electrode voltage clamp at room temperature (oocytes) or the whole-cell patch clamp technique at 35 degrees C (L929 cells). In oocytes, during moderate extracellular acidification (pHo = 6.4), current activation was not shifted on the voltage axis, the time course of current activation was unchanged, but tail current deactivation was dramatically accelerated. At pHo < 6.4, in addition to accelerating deactivation, the time course of activation was slower and the midpoint voltage of current activation was shifted to more positive values. Protons and Zn2+ accelerated the kinetics of deactivation with apparent Kd values about one order of magnitude lower than for tail current inhibition. For protons, the Kd values for the effect on tail current amplitude versus kinetics were, respectively, 1.8 microM (pKa = 5.8) and 0.1 microM (pKa = 7.0). In the presence of Zn2+, the corresponding Kd values were, respectively, 1.2 mM and 169 microM. In L929 cells, acidification to pHo = 6.4 did not shift the midpoint voltage of current activation and had no effect on the time course of current activation. Furthermore, the onset and recovery of inactivation were not affected. However, the acidification significantly accelerated tail current deactivation. We conclude that protons and Zn2+ directly interact with HERG channels and that the interaction results, preferentially, in the regulation of channel deactivation mechanism. PMID- 10388758 TI - Effect of gramicidin A on the dipole potential of phospholipid membranes. AB - The effect of channel-forming peptide gramicidin A on the dipole potential of phospholipid monolayers and bilayers has been studied. Surface pressure and surface potential isotherms of monolayers have been measured with a Langmuir trough equipped with a Wilhelmy balance and a surface potential meter (Kelvin probe). Gramicidin has been shown to shift pressure-area isotherms of phospholipids and to reduce their monolayer surface potentials. Both effects increase with the increase in gramicidin concentration and depend on the kind of phosphatidylcholine used. Application of the dual-wavelength ratiometric fluorescence method using the potential-sensitive dye RH421 has revealed that the addition of gramicidin A to dipalmitoylphosphatidylcholine liposomes leads to a decrease in the fluorescence ratio of RH421. This is similar to the effect of phloretin, which is known to decrease the dipole potential. The comparison of the concentration dependences of the fluorescence ratio for gramicidin and phloretin shows that gramicidin is as potent as phloretin in modifying the membrane dipole potential. PMID- 10388759 TI - Divalent cation-mediated interaction between cerebroside sulfate and cerebrosides: an investigation of the effect of structural variations of lipids by electrospray ionization mass spectrometry. AB - Divalent cations mediate a carbohydrate-carbohydrate association between the two major glycolipids, galactosylceramide (GalCer) and its sulfated form, cerebroside sulfate (CBS), of the myelin sheath. We have suggested that interaction between these glycolipids on apposed extracellular surfaces of myelin may be involved in the stability or function of this multilayered structure. A mutant mouse lacking galactolipids because of a disruption in the gene that encodes a galactosyltransferase forms myelin that initially appears relatively normal but is unstable. This myelin contains glucosylceramide (GlcCer) instead of GalCer. To better understand the role of GlcCer in myelin in this mutant, we have compared the ability of divalent cations to complex CBS (galactosyl form) with GlcCer or GalCer in methanol solution by using positive ion electrospray ionization mass spectrometry. Because both the alpha-hydroxylated fatty acid species (HFA) and the nonhydroxylated fatty acid species (NFA) of these lipids occur in myelin, we have also compared the HFA and NFA species. In addition to monomeric Ca2+ complexes of all three lipids and oligomeric Ca2+ complexes of both GalCer and GlcCer, Ca2+ also caused heterotypic complexation of CBS to both GalCer and GlcCer. The heterotypic complexes had the greatest stability of all oligomers formed and survived better at high declustering potentials. Complexes of CBS with GlcCer were less stable than those with GalCer. This was confirmed by using the free sugars and glycosides making up the carbohydrate headgroups of these lipids. HFA species of CBS and GalCer formed more stable complexes than NFA species, but hydroxylation of the fatty acid of GlcCer had no effect. The ability of GlcCer to also complex with CBS, albeit with lower stability, may allow GlcCer to partially compensate for the absence of GalCer in the mouse mutant. PMID- 10388760 TI - Modulation of nanotube formation by structural modifications of sphingolipids. AB - Galactosylceramides (GalCers) containing nervonoyl (24:1(Delta15(cis))) acyl chains have the capacity to assemble into nanotubular microstructures in excess water (. Biophys. J. 69:1976-1986). To define the structural parameters that modulate nanotube formation, GalCer derivatives were synthesized that contained cis monounsaturated acyl chains with the formula X:1((X-9)). X indicates the total acyl carbon number (24, 22, 20, or 18), and 1 indicates a single cis double bond, the location of which is designated by the superscript (X-9). Deep etching of freeze-fractured 24:1(Delta15(cis)) GalCer dispersions followed by replica production and transmission electron microscopic analysis confirmed nanotube morphology (25-30-nm diameter). Control experiments revealed that tubule formation was promoted by cooling through the main enthalpic phase transition coupled with repetitive freeze-thaw cycling. Imparting a negative charge to the sugar headgroup of 24:1(Delta15)GalCer via sulfate dramatically altered mesomorpholgy and resulted in myelinic-like, multilamellar structures. Removal of the sugar headgroup (24:1(Delta15)Cer) resulted in flattened cylindrical structures with a cochleate appearance. Compared to these large-scale changes in morphology, more subtle changes were induced by structural changes in the acyl chain of 24:1(Delta15)GalCer. 22:1(Delta13)GalCer dispersions consisted of long, smooth tubules (35-40-nm diameters) with a strong tendency to self-align into bundle-like aggregates. In contrast, the microstructures formed by 20:1(Delta11)GalCer resembled helical ribbons with a right-handed twist. Ribbon widths averaged 30-35 nm, with helical pitches of 80-90 nm. 18:1(Delta9)GalCer displayed a variety of morphologies, including large-diameter multilamellar cylinders and liposome-like structures, as well as stacked, plate-like arrays. The results are discussed within the context of current theories of lipid tubule formation. PMID- 10388761 TI - Anisotropic motion of cholesterol in oriented DPPC bilayers studied by quasielastic neutron scattering: the liquid-ordered phase. AB - Quasielastic neutron scattering (QENS) at two energy resolutions (1 and 14 microeV) was employed to study high-frequency cholesterol motion in the liquid ordered phase (lo-phase) of oriented multilayers of dipalmitoylphosphatidylcholine at three temperatures: T = 20 degrees C, T = 36 degrees C, and T = 50 degrees C. We studied two orientations of the bilayer stack with respect to the incident neutron beam. This and the two energy resolutions for each orientation allowed us to determine the cholesterol dynamics parallel to the normal of the membrane stack and in the plane of the membrane separately at two different time scales in the GHz range. We find a surprisingly high, model independent motional anisotropy of cholesterol within the bilayer. The data analysis using explicit models of molecular motion suggests a superposition of two motions of cholesterol: an out-of-plane diffusion of the molecule parallel to the bilayer normal combined with a locally confined motion within the bilayer plane. The rather high amplitude of the out-of-plane diffusion observed at higher temperatures (T >/= 36 degrees C) strongly suggests that cholesterol can move between the opposite leaflets of the bilayer while it remains predominantly confined within its host monolayer at lower temperatures (T = 20 degrees C). The locally confined in-plane cholesterol motion is dominated by discrete, large angle rotational jumps of the steroid body rather than a quasicontinous rotational diffusion by small angle jumps. We observe a significant increase of the rotational jump rate between T = 20 degrees C and T = 36 degrees C, whereas a further temperature increase to T = 50 degrees C leaves this rate essentially unchanged. PMID- 10388762 TI - Physicochemical characterization and purification of cationic lipoplexes. AB - Cationic lipid-nucleic acid complexes (lipoplexes) consisting of dioleoyltrimethylammoniumpropane (DOTAP) liposomes and plasmid DNA were prepared at various charge ratios (cationic group to nucleotide phosphate), and the excess component was separated from the lipoplex. We measured the stoichiometry of the lipoplex, noted its colloidal properties, and observed its morphology and structure by electron microscopy. The colloidal properties of the lipoplexes were principally determined by the cationic lipid/DNA charge ratio and were independent of the lipid composition. In lipoplexes, the lipid membranes as observed in freeze-fracture electron microscopy were deformed into high-radius-of curvature features whose characteristics depended on the lipid composition. Lipoplexes prepared at a threefold or greater excess of either DOTAP or DNA could be resolved into complexes with a defined stoichiometry and the excess component by sedimentation to equilibrium on sucrose gradients. The separated, positively charged complex retained high transfection activity and had reduced toxicity. The negatively charged lipoplex showed increased transfection activity compared to the starting mixture. In cryoelectron micrographs the positively charged complex was spherical and contained a condensed but indistinct interior structure. In contrast, the separated negatively charged lipoplexes had a prominent internal 5.9 +/- 0.1-nm periodic feature with material projecting as spikes from the spherical structure into the solution. It is likely that these two lipoplexes represent structures with different lipid and DNA packing. PMID- 10388763 TI - Structural changes in the actin-myosin cross-bridges associated with force generation induced by temperature jump in permeabilized frog muscle fibers. AB - Structural changes induced by Joule temperature jumps (T-jumps) in frog muscle fibers were monitored using time-resolved x-ray diffraction. Experiments made use of single, permeabilized fibers that were fully activated after slight cross linking with 1-ethyl-3-[3-dimethylamino)propyl]carbodiimide to preserve their structural order. After T-jumps from 5-6 to approximately 17 degrees C and then on to approximately 30 degrees C, tension increased by a factor of 1.51 and 1.84, respectively, whereas fiber stiffness did not change with temperature. The tension rise was accompanied by a decrease in the intensity of the (1, 0) equatorial x-ray reflection by 15 and 26% (at approximately 17 and approximately 30 degrees C) and by an increase in the intensity of the M3 myosin reflection by 20% and 41%, respectively. The intensity of the (1,1) equatorial reflection increased slightly. The peak of the intensity on the 6th actin layer line shifted toward the meridian with temperature. The intensity of the 1st actin layer line increased from 12% (of its rigor value) at 5-6 degrees C to 36% at approximately 30 degrees C, so that the fraction of the cross-bridges labeling the actin helix estimated from this intensity increased proportionally to tension from approximately 35% at 5-6 degrees C to approximately 60% at approximately 30 degrees C. This suggests that force is generated during a transition of nonstereo specifically attached myosin cross-bridges to a stereo-specific binding state. PMID- 10388765 TI - The ADP release step of the smooth muscle cross-bridge cycle is not directly associated with force generation. AB - When smooth muscle myosin subfragment 1 (S1) is bound to actin filaments in vitro, the light chain domain tilts upon release of MgADP, producing a approximately 3.5-nm axial motion of the head-rod junction (Whittaker et al., 1995. Nature. 378:748-751). If this motion contributes significantly to the power stroke, rigor tension of smooth muscle should decrease substantially in response to cross-bridge binding of MgADP. To test this prediction, we monitored mechanical properties of permeabilized strips of chicken gizzard muscle in rigor and in the presence of MgADP. For comparison, we also tested psoas and soleus muscle fibers. Any residual bound ADP was minimized by incubation in Mg2+-free rigor solution containing 15 mM EDTA. The addition of 2 mM MgADP, while keeping ionic strength and free Mg2+ concentration constant, resulted in a slight increase in rigor tension in both gizzard and soleus muscles, but a decrease in psoas muscle. In-phase stiffness monitored during small (<0.1%) 500-Hz sinusoidal length oscillations decreased in all three muscle types when MgADP was added. The changes in force and stiffness with the addition of MgADP were similar at ionic strengths from 50 to 200 mM and were reversible. The results with gizzard muscle were similar after thiophosphorylation of the regulatory light chain of myosin. These results suggest that the axial motion of smooth muscle S1 bound to actin, upon dissociation of MgADP, is not associated with force generation. The difference between the present mechanical data and previous structural studies of smooth S1 may be explained if geometrical constraints of the intact contractile filament array alter the motions of the myosin heads. PMID- 10388764 TI - Divalent cation-, nucleotide-, and polymerization-dependent changes in the conformation of subdomain 2 of actin. AB - Conformational changes in subdomain 2 of actin were investigated using fluorescence probes dansyl cadaverine (DC) or dansyl ethylenediamine (DED) covalently attached to Gln41. Examination of changes in the fluorescence emission spectra as a function of time during Ca2+/Mg2+ and ATP/ADP exchange at the high affinity site for divalent cation-nucleotide complex in G-actin confirmed a profound influence of the type of nucleotide but failed to detect a significant cation-dependent difference in the environment of Gln41. No significant difference between Ca- and Mg-actin was also seen in the magnitude of the fluorescence changes resulting from the polymerization of these two actin forms. Evidence is presented that earlier reported cation-dependent differences in the conformation of the loop 38-52 may be related to time-dependent changes in the conformation of subdomain 2 in DED- or DC-labeled G-actin, accelerated by substitution of Mg2+ for Ca2+ in CaATP-G-actin and, in particular, by conversion of MgATP- into MgADP-G-actin. These spontaneous changes are associated with a denaturation-driven release of the bound nucleotide that is promoted by two effects of DED or DC labeling: lowered affinity of actin for nucleotide and acceleration of ATP hydrolysis on MgATP-G-actin that converts it into a less stable MgADP form. Evidence is presented that the changes in the environment of Gln41 accompanying actin polymerization result in part from the release of Pi after the hydrolysis of ATP on the polymer. A similarity of this change to that accompanying replacement of the bound ATP with ADP in G-actin is discussed. PMID- 10388767 TI - Self-aggregation of DNA oligomers with XGG trinucleotide repeats: kinetic and atomic force microscopy measurements. AB - Turbidity measurements via absorbance monitoring at 320 nm were employed to obtain autocatalytic-like kinetic profiles of K+-induced aggregate formation of d(XGG)4 and some related oligomers, where X = A, C, G, and T. At least 1 M KCl is needed to observe the turbidity-measurable aggregation at pH 8, and the relative propensity for aggregate formation is shown to follow the order d(GGG)4 > d(AGG)4 approximately d(TGG)4 >> d(CGG)4. The presence of Mg2+ greatly facilitates and dramatically reduces the amount of K+ required to initiate aggregation and significantly enhances the thermal stabilities of the aggregates. Replacement of K+ by Na+ fails to induce a similar phenomenon. The Psi-type CD characteristics of aggregates are strongly dependent on the sequence and ionic conditions. Despite their ease of aggregate formation, oligomers with AGG trinucleotide repeats fail to exhibit Psi-CD formation. The propensity for aggregation is greatly affected by the chain length, with oligomers of four repeats being most facile. Appending X base at the 3' end of d(GGXGGXGGXGG) appears to provide a greater hindrance to aggregation than at the 5' end. Atomic force microscopic images support some of these findings and reveal the morphologies of these aggregates. The presence of MgCl2 in solutions appears to considerably elongate the K+-induced aggregates. PMID- 10388766 TI - Unexpected BII conformer substate population in unoriented hydrated films of the d(CGCGAATTCGCG)2 dodecamer and of native B-DNA from salmon testes. AB - Conformational substates of B-DNA had been observed so far in synthetic oligonucleotides but not in naturally occurring highly polymeric B-DNA. Our low temperature experiments show that native B-DNA from salmon testes and the d(CGCGAATTCGCG)2 dodecamer have the same BI and BII substates. Nonequilibrium distribution of conformer population was generated by quenching hydrated unoriented films to 200 K, and isothermal structural relaxation toward equilibrium by interconversion of substates was followed by Fourier transform infrared spectroscopy. BI interconverts into BII on isothermal relaxation at 200 K, whereas on slow cooling from ambient temperature, BII interconverts into BI. Our estimation of the dodecamer's BI-to-BII conformer substate population by curve resolution of the symmetrical stretching vibration of the ionic phosphate is 2.4 +/- 0.5 to 1 at 200 K, and it is 1.3 +/- 0.5 to 1 between 270 and 290 K. Pronounced spectral changes upon BI-to-BII interconversion are consistent with base destacking coupled with migration of water from ionic phosphate toward the phosphodiester and sugar moieties. Nonspecific interaction of proteins with the DNA backbone could become specific by induced-fit-type interactions with either BI or BII backbone conformations. This suggests that the BI-to-BII substate interconversion could be a major contributor to the protein recognition process. PMID- 10388768 TI - Exciton delocalization in the B808-866 antenna of the green bacterium Chloroflexus aurantiacus as revealed by ultrafast pump-probe spectroscopy. AB - A model of pigment organization in the B808-866 bacteriochlorophyll a antenna of the green photosynthetic bacterium Chloroflexus aurantiacus based on femtosecond pump-probe studies is proposed. The building block of the antenna was assumed to be structurally similar to that of the B800-850 light-harvesting 2 (LH2) antenna of purple bacteria and to have the form of two concentric rings of N strongly coupled BChl866 pigments and of N/2 weakly coupled BChl808 monomers, where N = 24 or 32. We have shown that the Qy transition dipoles of BChl808 and BChl866 molecules form the angles 43 degrees +/- 3 degrees and 8 degrees +/- 4 degrees, respectively, with the plane of the corresponding rings. Using the exciton model, we have obtained a quantitative fit of the pump-probe spectra of the B866 and B808 bands. The anomalously high bleaching value of the B866 band with respect to the B808 monomeric band provided the direct evidence for a high degree of exciton delocalization in the BChl866 ring antenna. The coherence length of the steady state exciton wave packet corresponds to five or six BChl866 molecules at room temperature. PMID- 10388769 TI - Long-distance effects of site-directed mutations on backbone conformation in bacteriorhodopsin from solid state NMR of [1-13C]Val-labeled proteins. AB - We have recorded 13C cross-polarization-magic angle spinning and dipolar decoupled-magic angle spinning NMR spectra of [1-13C]Val-labeled wild-type bacteriorhodopsin (bR), and the V49A, V199A, T46V, T46V/V49A, D96N, and D85N mutants, in order to study conformational changes of the backbone caused by site directed mutations along the extracellular surface and the cytoplasmic half channel. On the basis of spectral changes in the V49A and V199A mutants, and upon specific cleavage by chymotrypsin, we assigned the three well-resolved 13C signals observed at 172.93, 172.00, and 171. 11 ppm to [1-13C]Val 69, Val 49, and Val 199, respectively. The local conformations of the backbone at these residues are revealed by the conformation-dependent 13C chemical shifts. We find that at the ambient temperature of these measurements Val 69 is not in a beta-sheet, in spite of previous observations by electron microscopy and x-ray diffraction at cryogenic temperatures, but in a flexible turn structure that undergoes conformational fluctuation. Results with the T46V mutant suggest that there is a long-distance effect on backbone conformation between Thr 46 and Val 49. From the spectra of the D85N and E204Q mutants there also appears to be coupling between Val 49 and Asp 85 and between Asp 85 and Glu 204, respectively. In addition, the T2 measurement indicates conformational interaction between Asp 96 and extracellular surface. The protonation of Asp 85 in the photocycle therefore might induce changes in conformation or dynamics, or both, throughout the protein, from the extracellular surface to the side chain of Asp 96. PMID- 10388770 TI - NMR studies of electrostatic potential distribution around biologically important molecules. AB - A new experimental approach has been developed to study the distribution of local electrostatic potential around specific protons in biologically important molecules. The approach is the development of a method denoted as "spin label/spin probe," which was proposed by one of us (. Mol. Biol. 6:498-507). The proposed method is based upon the quantitative measurement of the contribution of differently charged nitroxide probes to the spin lattice relaxation rate (1/T1) of protons in the molecule of interest, followed by calculation of local electrostatic potential using the classical Debye equation. In parallel, the theoretical calculation of potential distribution with the use of the MacSpartan Plus 1.0 program has been performed. Application of the method to solutions of simple organic molecules (aliphatic and aromatic alcohols, aliphatic carboxylates (propionate anion), and protonated ethyl amine and imidazole) allowed us to estimate the effective potential around the molecules under investigation. These were found to be in good agreement with theoretically expected values. This technique was then applied to zwitterionic amino acids bearing neutral and charged side chains (glycine, lysine, histidine, and aspartic acid). The reliability of the general approach is proved by the data presented in this paper. Application of this new methodology can afford insight into the biochemical significance of electrostatic effects in biological systems. PMID- 10388771 TI - Evolution of the internal dynamics of two globular proteins from dry powder to solution. AB - Myoglobin and lysozyme picosecond internal dynamics in solution is compared to that in hydrated powders by quasielastic incoherent neutron scattering. This technique is sensitive to the motions of the nonexchangeable hydrogen atoms in a sample. Because these are homogeneously distributed throughout the protein structure, the average dynamics of the protein is described. We first propose an original data treatment to deal with the protein global motions in the case of solution samples. The validity of this treatment is checked by comparison with classical measurements of the diffusion constants. The evolution with the scattering vector of the width and relative contribution of the quasielastic component was then used to derive information on the amount of local diffusive motions and their characteristic average relaxation time. From dry powder to coverage by one water layer, the surface side chains progressively acquire the possibility to diffuse locally. On subsequent hydration, the main effect of water is to improve the rate of these diffusive motions. Motions with higher average amplitude occur in solution, about three times more than for a hydrated powder at complete coverage, with a shorter average relaxation time, approximately 4.5 ps compared to 9.4 ps for one water monolayer. PMID- 10388772 TI - Reversible thermal denaturation of human FGF-1 induced by low concentrations of guanidine hydrochloride. AB - Human acidic fibroblast growth factor (FGF-1) is a powerful mitogen and angiogenic factor with an apparent melting temperature (Tm) in the physiological range. FGF-1 is an example of a protein that is regulated, in part, by stability based mechanisms. For example, the low Tm of FGF-1 has been postulated to play an important role in the unusual endoplasmic reticulum-independent secretion of this growth factor. Despite the close relationship between function and stability, accurate thermodynamic parameters of unfolding for FGF-1 have been unavailable, presumably due to effects of irreversible thermal denaturation. Here we report the determination of thermodynamic parameters of unfolding (DeltaH, DeltaG, and DeltaCp) for FGF-1 using differential scanning calorimetry (DSC). The thermal denaturation is demonstrated to be two-state and reversible upon the addition of low concentrations of added guanidine hydrochloride (GuHCl). DeltaG values from the DSC studies are in excellent agreement with values from isothermal GuHCl denaturation monitored by fluorescence and circular dichroism (CD) spectroscopy. Furthermore, the results indicate that irreversible denaturation is closely associated with the formation of an unfolding intermediate. GuHCl appears to promote reversible two-state denaturation by initially preventing aggregation of this unfolding intermediate, and at subsequently higher concentrations, by preventing formation of the intermediate. PMID- 10388773 TI - The quantum mixed-spin heme state of barley peroxidase: A paradigm for class III peroxidases. AB - Electronic absorption and resonance Raman (RR) spectra of the ferric form of barley grain peroxidase (BP 1) at various pH values, at both room temperature and 20 K, are reported, together with electron paramagnetic resonance spectra at 10 K. The ferrous forms and the ferric complex with fluoride have also been studied. A quantum mechanically mixed-spin (QS) state has been identified. The QS heme species coexists with 6- and 5-cHS hemes; the relative populations of these three spin states are found to be dependent on pH and temperature. However, the QS species remains in all cases the dominant heme spin species. Barley peroxidase appears to be further characterized by a splitting of the two vinyl stretching modes, indicating that the vinyl groups are differently conjugated with the porphyrin. An analysis of the currently available spectroscopic data for proteins from all three peroxidase classes suggests that the simultaneous occurrence of the QS heme state as well as the splitting of the two vinyl stretching modes is confined to class III enzymes. The former point is discussed in terms of the possible influences of heme deformations on heme spin state. It is found that moderate saddling alone is probably not enough to cause the QS state, although some saddling may be necessary for the QS state. PMID- 10388774 TI - Molecular dynamics of microbial lipases as determined from their intrinsic tryptophan fluorescence. AB - We have studied the intrinsic tryptophan fluorescence of the lipases from Chromobacterium viscosum (CVL), Pseudomonas species (PSL), and Rhizopus oryzae (ROL) in aqueous buffer, zwitterionic detergent micelles, and isopropanol-water mixtures. It was the purpose of this study to obtain information about biophysical properties of the respective enzymes under conditions that modulate enzyme activities and stereoselectivities to a significant extent. According to their decay-associated emission spectra, CVL tryptophans are located in the hydrophobic interior of the protein. In contrast, the PSL and ROL tryptophans are probably confined to the core and the surface of the lipase. From the tryptophan lifetime distributions it can be concluded that the conformation of CVL is not much affected by detergent or organic solvent (isopropanol). Accordingly, CVL is enzymatically active in these systems and most active in the presence of isopropanol. In contrast, ROL and PSL show high conformational mobility, depending on the solvent, because their lifetime distributions are very different in the presence and absence of detergent or isopropanol. Time-resolved anisotropy studies provided evidence that the lipases exhibit very high internal molecular flexibility. This peculiar feature of lipases is perhaps the key to the great differences in activity and stereoselectivity observed in different reaction media. Furthermore, information about self-association of the lipases in different solvents could be obtained. PSL, but not CVL and ROL, forms aggregates in water. Lipase aggregation can be reversed by the addition of detergent or isopropanol, which competes for the hydrophobic surface domains of this protein. This dissociation could efficiently contribute to the increase in lipase activity in the presence of a detergent or isopropanol. PMID- 10388776 TI - Separation of submicron bioparticles by dielectrophoresis. AB - Submicron particles such as latex spheres and viruses can be manipulated and characterized using dielectrophoresis. By the use of appropriate microelectrode arrays, particles can be trapped or moved between regions of high or low electric fields. The magnitude and direction of the dielectrophoretic force on the particle depends on its dielectric properties, so that a heterogeneous mixture of particles can be separated to produce a more homogeneous population. In this paper the controlled separation of submicron bioparticles is demonstrated. With electrode arrays fabricated using direct write electron beam lithography, it is shown that different types of submicron latex spheres can be spatially separated. The separation occurs as a result of differences in magnitude and/or direction of the dielectrophoretic force on different populations of particles. These differences arise mainly because the surface properties of submicron particles dominate their dielectrophoretic behavior. It is also demonstrated that tobacco mosaic virus and herpes simplex virus can be manipulated and spatially separated in a microelectrode array. PMID- 10388777 TI - Protein adhesion force dynamics and single adhesion events. AB - Using the manipulation force microscope, a novel atomic force microscope, the adhesion forces of bovine serum albumin, myoglobin, ferritin, and lysozyme proteins to glass and polystyrene substrates were characterized by following the force necessary to displace an adsorbed protein-covered microsphere over several orders of magnitude in time. This force was consistent with a power law with exponent a = 0.37 +/- 0.03 on polystyrene, indicating that there is no typical time scale for adhesion on this substrate. On glass, the rate of adhesion depended strongly on protein charge. Forces corresponding to single protein adhesion events were identified. The typical rupture force of a single lysozyme, ferritin, bovine serum albumin, and myoglobin protein adhering to glass was estimated to be 90 +/- 10 pN, 115 +/- 13 pN, 277 +/- 44 pN, and 277 +/- 44 pN, respectively, using a model of the experimental system. These forces, as well as the force amplitudes on hydrophobic polystyrene, correlate with protein stiffness. PMID- 10388775 TI - Molecular dynamics simulations of protein-tyrosine phosphatase 1B. I. ligand induced changes in the protein motions. AB - Activity of enzymes, such as protein tyrosine phosphatases (PTPs), is often associated with structural changes in the enzyme, resulting in selective and stereospecific reactions with the substrate. To investigate the effect of a substrate on the motions occurring in PTPs, we have performed molecular dynamics simulations of PTP1B and PTP1B complexed with a high-affinity peptide DADEpYL, where pY stands for phosphorylated tyrosine. The peptide sequence is derived from the epidermal growth factor receptor (EGFR988-993). Simulations were performed in water for 1 ns, and the concerted motions in the protein were analyzed using the essential dynamics technique. Our results indicate that the predominately internal motions in PTP1B occur in a subspace of only a few degrees of freedom. Upon substrate binding, the flexibility of the protein is reduced by approximately 10%. The largest effect is found in the protein region, where the N terminal of the substrate is located, and in the loop region Val198-Gly209. Displacements in the latter loop are associated with the motions in the WPD loop, which contains a catalytically important aspartic acid. Estimation of the pKa of the active-site cysteine along the trajectory indicates that structural inhomogeneity causes the pKa to vary by approximately +/-1 pKa unit. In agreement with experimental observations, the active-site cysteine is negatively charged at physiological pH. PMID- 10388778 TI - Atomic force microscopy of the submolecular architecture of hydrated ocular mucins. AB - High-resolution atomic force microscopy has been applied to the imaging of intact human ocular mucins in a near-physiological buffer. The mucins displayed a range of lengths from several hundred nanometers to several microns. By varying the ionic composition of the imaging environment, it was possible to image molecules rigidly fixed to the substrate and the motion of single molecules across the substrate. From static molecular images, high-resolution line profiles show a variation of up to +/-0.75 nm in thickness along the molecule. This variation is localized in regions of several tens of nanometers. It is interpreted in terms of the varying glycosylation along the mucin and is consistent with the known size of oligosaccharides in ocular mucins. The dynamic images indicate the possibility of following mucin interactions in situ. PMID- 10388779 TI - Diffusion of macromolecules in agarose gels: comparison of linear and globular configurations. AB - The diffusion coefficients (D) of different types of macromolecules (proteins, dextrans, polymer beads, and DNA) were measured by fluorescence recovery after photobleaching (FRAP) both in solution and in 2% agarose gels to compare transport properties of these macromolecules. Diffusion measurements were conducted with concentrations low enough to avoid macromolecular interactions. For gel measurements, diffusion data were fitted according to different theories: polymer chains and spherical macromolecules were analyzed separately. As chain length increases, diffusion coefficients of DNA show a clear shift from a Rouse like behavior (DG congruent with N0-0.5) to a reptational behavior (DG congruent with N0-2.0). The pore size, a, of a 2% agarose gel cast in a 0.1 M PBS solution was estimated. Diffusion coefficients of the proteins and the polymer beads were analyzed with the Ogston model and the effective medium model permitting the estimation of an agarose gel fiber radius and hydraulic permeability of the gels. Not only did flexible macromolecules exhibit greater mobility in the gel than did comparable-size rigid spherical particles, they also proved to be a more useful probe of available space between fibers. PMID- 10388780 TI - The photon counting histogram in fluorescence fluctuation spectroscopy. AB - Fluorescence correlation spectroscopy (FCS) is generally used to obtain information about the number of fluorescent particles in a small volume and the diffusion coefficient from the autocorrelation function of the fluorescence signal. Here we demonstrate that photon counting histogram (PCH) analysis constitutes a novel tool for extracting quantities from fluorescence fluctuation data, i.e., the measured photon counts per molecule and the average number of molecules within the observation volume. The photon counting histogram of fluorescence fluctuation experiments, in which few molecules are present in the excitation volume, exhibits a super-Poissonian behavior. The additional broadening of the PCH compared to a Poisson distribution is due to fluorescence intensity fluctuations. For diffusing particles these intensity fluctuations are caused by an inhomogeneous excitation profile and the fluctuations in the number of particles in the observation volume. The quantitative relationship between the detected photon counts and the fluorescence intensity reaching the detector is given by Mandel's formula. Based on this equation and considering the fluorescence intensity distribution in the two-photon excitation volume, a theoretical expression for the PCH as a function of the number of molecules in the excitation volume is derived. For a single molecular species two parameters are sufficient to characterize the histogram completely, namely the average number of molecules within the observation volume and the detected photon counts per molecule per sampling time epsilon. The PCH for multiple molecular species, on the other hand, is generated by successively convoluting the photon counting distribution of each species with the others. The influence of the excitation profile upon the photon counting statistics for two relevant point spread functions (PSFs), the three-dimensional Gaussian PSF conventionally employed in confocal detection and the square of the Gaussian-Lorentzian PSF for two photon excitation, is explicitly treated. Measured photon counting distributions obtained with a two-photon excitation source agree, within experimental error with the theoretical PCHs calculated for the square of a Gaussian-Lorentzian beam profile. We demonstrate and discuss the influence of the average number of particles within the observation volume and the detected photon counts per molecule per sampling interval upon the super-Poissonian character of the photon counting distribution. PMID- 10388781 TI - Atomic force microscopy imaging of DNA covalently immobilized on a functionalized mica substrate. AB - A procedure for covalent binding of DNA to a functionalized mica substrate is described. The approach is based on photochemical cross-linking of DNA to immobilized psoralen derivatives. A tetrafluorphenyl (TFP) ester of trimethyl psoralen (trioxalen) was synthesized, and the procedure to immobilize it onto a functionalized aminopropyl mica surface (AP-mica) was developed. DNA molecules were cross-linked to trioxalen moieties by UV irradiation of complexes. The steps of the sample preparation procedure were analyzed with x-ray photoelectron spectroscopy (XPS). Results from XPS show that an AP-mica surface can be formed by vapor phase deposition of silane and that this surface can be derivatized with trioxalen. The derivatized surface is capable of binding of DNA molecules such that, after UV cross-linking, they withstand a thorough rinsing with SDS. Observations with atomic force microscopy showed that derivatized surfaces remain smooth, so DNA molecules are easily visualized. Linear and circular DNA molecules were photochemically immobilized on the surface. The molecules are distributed over the surface uniformly, indicating rather even modification of AP-mica with trioxalen. Generally, the shapes of supercoiled molecules electrostatically immobilized on AP-mica and those photocross-linked on trioxalen-functionalized surfaces remain quite similar. This suggests that UV cross-linking does not induce formation of a noticeable number of single-stranded breaks in DNA molecules. PMID- 10388782 TI - Caffeine interaction with fluorescent calcium indicator dyes. AB - We report that caffeine, in millimolar concentrations, interacts strongly with four common calcium indicator dyes: mag-fura-2, magnesium green, fura-2, and fluo 3. Fluorescence intensities are either noticeably enhanced (mag-fura-2, fura-2) or diminished (magnesium green, fluo-3). The caffeine-induced changes in the fluorescence spectra are clearly distinct from those of metal ion binding at the indicator chelation sites. Binding affinities for calcium of either mag-fura-2 or magnesium green increased only slightly in the presence of caffeine. Caffeine also alters the fluorescence intensities of two other fluorescent dyes lacking a chelation site, fluorescein and sulforhodamine 101, implicating the fluorophore itself as the interaction site for caffeine. In the absence of caffeine, variation of solution hydrophobicity by means of water/dioxane mixtures yielded results similar to those for caffeine. These observations suggest that hydrophobic substances, in general, can alter dye fluorescence in a dye-specific manner. For the particular case of caffeine, and perhaps other commonly used pharmacological agents, the dye interactions can seriously distort fluorescence measurements of intracellular ion concentrations with metal indicator dyes. PMID- 10388783 TI - Mechanical anchoring strength of L-selectin, beta2 integrins, and CD45 to neutrophil cytoskeleton and membrane. AB - The strength of anchoring of transmembrane receptors to cytoskeleton and membrane is important in cell adhesion and cell migration. With micropipette suction, we applied pulling forces to human neutrophils adhering to latex beads that were coated with antibodies to CD62L (L-selectin), CD18 (beta2 integrins), or CD45. In each case, the adhesion frequency between the neutrophil and bead was low, and our Monte Carlo simulation indicates that only a single bond was probably involved in every adhesion event. When the adhesion between the neutrophil and bead was ruptured, it was very likely that receptors were extracted from neutrophil surfaces. We found that it took 1-2 s to extract an L-selectin at a force range of 25-45 pN, 1-4 s to extract a beta2 integrin at a force range of 60 130 pN, and 1-11 s to extract a CD45 at a force range of 35-85 pN. Our results strongly support the conclusion that, during neutrophil rolling, L-selectin is unbound from its ligand when the adhesion between neutrophils and endothelium is ruptured. PMID- 10388784 TI - Temperature dependence of switching of the bacterial flagellar motor by the protein CheY(13DK106YW). AB - The behavior of the bacterium Escherichia coli is controlled by switching of the flagellar rotary motor between the two rotational states, clockwise (CW) and counterclockwise (CCW). The molecular mechanism for switching remains unknown, but binding of the response regulator CheY-P to the motor component FliM enhances CW rotation. This effect is mimicked by the unphosphorylated double mutant CheY13DK106YW (CheY**). To learn more about switching, we measured the fraction of time that a motor spends in the CW state (the CW bias) at different concentrations of CheY** and at different temperatures. From the CW bias, we computed the standard free energy change of switching. In the absence of CheY, this free energy change is a linear function of temperature (. Biophys. J. 71:2227-2233). In the presence of CheY**, it is nonlinear. However, the data can be fit by models in which binding of each molecule of CheY** shifts the difference in free energy between CW and CCW states by a fixed amount. The shift increases linearly from approximately 0.3kT per molecule at 5 degrees C to approximately 0.9kT at 25 degrees C, where k is Boltzmann's constant and T is 289 Kelvin (= 16 degrees C). The entropy and enthalpy contributions to this shift are about -0. 031kT/ degrees C and 0.10kT, respectively. PMID- 10388785 TI - Dynamin: possible mechanism of "Pinchase" action. AB - Dynamin is a GTPase playing an essential role in ubiquitous intra cellular processes involving separation of vesicles from plasma membranes and membranes of cellular compartments. Recent experimental progress (. Cell. 93:1021-1029;. Cell. 94:131-141) has made it possible to attempt to understand the action of dynamin in physical terms. Dynamin molecules are shown to bind to a lipid membrane, to self-assemble into a helicoidal structure constricting the membrane into a tubule, and, as a result of GTP hydrolysis, to mediate fission of this tubule (). In a similar way, dynamin is supposed to mediate fission of a neck connecting an endocytic bud and the plasma membrane, i.e., to complete endocytosis. We suggest a mechanism of this "pinchase" action of dynamin. We propose that, as a result of GTP hydrolysis, dynamin undergoes a conformational change manifested in growth of the pitch of the dynamin helix. We show that this gives rise to a dramatic change of shape of the tubular membrane constricted inside the helix, resulting in a local tightening of the tubule, which is supposed to promote its fission. We treat this model in terms of competing elasticities of the dynamin helix and the tubular membrane and discuss the predictions of the model in relation to the previous views on the mechanism of dynamin action. PMID- 10388786 TI - Determination of time-dependent inositol-1,4,5-trisphosphate concentrations during calcium release in a smooth muscle cell. AB - The level of [InsP3]cyt required for calcium release in A7r5 cells, a smooth muscle cell line, was determined by a new set of procedures using quantitative confocal microscopy to measure release of InsP3 from cells microinjected with caged InsP3. From these experiments, the [InsP3]cyt required to evoke a half maximal calcium response is 100 nM. Experiments with caged glycerophosphoryl-myo inositol 4, 5-bisphosphate (GPIP2), a slowly metabolized analogue of InsP3, gave a much slower recovery and a half-maximal response of an order of magnitude greater than InsP3. Experimental data and highly constrained variables were used to construct a mathematical model of the InsP3-dependent [Ca2+]cyt changes; the resulting simulations show high fidelity to experiment. Among the elements considered in constructing this model were the mechanism of the InsP3-receptor, InsP3 degradation, calcium buffering in the cytosol, and refilling of the ER stores via sarcoplasmic endoplasmic reticulum ATPase (SERCA) pumps. The model predicts a time constant of 0.8 s for InsP3 degradation and 13 s for GPIP2. InsP3 degradation was found to be a prerequisite for [Ca2+]cyt recovery to baseline levels and is therefore critical to the pattern of the overall [Ca2+]cyt signal. Analysis of the features of this model provides insights into the individual factors controlling the amplitude and shape of the InsP3-mediated calcium signal. PMID- 10388788 TI - The mechanisms of lesion genesis in multiple sclerosis? PMID- 10388787 TI - Regulation of airway ciliary activity by Ca2+: simultaneous measurement of beat frequency and intracellular Ca2+. AB - Airway ciliary activity is influenced by [Ca2+]i, but this mechanism is not fully understood. To investigate this relationship, ciliary activity and [Ca2+]i were measured simultaneously from airway epithelial ciliated cells. Ciliary beat frequency was determined, for each beat cycle, with phase-contrast optics and high-speed video imaging (at 240 images s-1) and correlated with [Ca2+]i determined, at the ciliary base, by fast imaging (30 images s-1) of fura-2 fluorescence. As a mechanically induced intercellular Ca2+ wave propagated through adjacent cells, [Ca2+]i was elevated from a baseline concentration of 45 to 100 nM, to a peak level of up to 650 nM. When the Ca2+ wave reached the ciliary base, the beat frequency rapidly increased, within a few beat cycles, from a basal rate of 6.4 to 11.6 Hz at 20-23 degrees C, and from 17.2 to 26.7 Hz at 37 degrees C. Changes in [Ca2+]i, above 350 nM, had no effect on the maximum beat frequency. We suggest that airway ciliary beat frequency is 1) controlled by a low range of [Ca2+]i acting directly at an axonemal site at the ciliary base and 2) that a maximum frequency is induced by a change in [Ca2+]i of approximately 250-300 nM. PMID- 10388789 TI - Neurological disorders of micturition and their treatment. AB - An overview of the current concepts of the neurological control of the bladder is given, based on laboratory experiments and PET scanning studies in human subjects. This is followed by a description of the various causes of the neurogenic bladder, discussed in a hierarchical order starting with cortical lesions and descending through the basal ganglia and brainstem, spinal cord, conus and cauda equina to disorders of peripheral innervation. Then follows a description of the condition of isolated urinary retention in young women. The article concludes with a review of the methods available for treating neurogenic bladder disorders. These are largely medical but brief mention of appropriate surgical procedures is made. PMID- 10388791 TI - The perceptual consequences of visual loss: 'positive' pathologies of vision. AB - Fifty patients with visual hallucinations and illusions secondary to degenerative eye disease reported remarkably stereotyped experiences. Questionnaire responses revealed five previously recognized categories of pathological vision (perseveration, illusory visual spread, polyopia, prosopometamorphopsia and micro/macropsia) and three novel categories (tessellopsia, hyperchromatopsia and dendropsia). Identical pathologies of vision occur in a range of clinical and experimental settings, suggesting that they reflect fundamental visual processes. The known neurophysiology of the visual cortex helps explain the phenomenology of the experiences and provides the basis for a neurobiologically based classification of positive and negative visual perceptual disorders. PMID- 10388790 TI - Clinical genetics of familial progressive supranuclear palsy. AB - Recent studies have shown that progressive supranuclear palsy (PSP) could be inherited, but the pattern of inheritance and the spectrum of the clinical findings in relatives are unknown. We here report 12 pedigrees, confirmed by pathology in four probands, with familial PSP. Pathological diagnosis was confirmed according to recently reported internationally agreed criteria. The spectrum of the clinical phenotypes in these families was variable including 34 typical cases of PSP (12 probands plus 22 secondary cases), three patients with postural tremor, three with dementia, one with parkinsonism, two with tremor, dystonia, gaze palsy and tics, and one with gait disturbance. The presence of affected members in at least two generations in eight of the families and the absence of consanguinity suggests autosomal dominant transmission with incomplete penetrance. We conclude that hereditary PSP is more frequent than previously thought and that the scarcity of familial cases may be related to a lack of recognition of the variable phenotypic expression of the disease. PMID- 10388793 TI - Mechanisms underlying gait disturbance in Parkinson's disease: a single photon emission computed tomography study. AB - Single photon emission computed tomography was used to evaluate regional cerebral blood flow changes during gait on a treadmill in 10 patients with Parkinson's disease and 10 age-matched controls. The subjects were injected with [99mTc]hexamethyl-propyleneamine oxime twice: while walking on the treadmill, which moved at a steady speed, and while lying on a bed with their eyes open. On the treadmill, all subjects walked at the same speed with their preferred stride length. The patients showed typical hypokinetic gait with higher cadence and smaller stride length than the controls. In the controls, a gait-induced increase in brain activity was observed in the medial and lateral premotor areas, primary sensorimotor areas, anterior cingulate contex, superior parietal cortex, visual cortex, dorsal brainstem, basal ganglia and cerebellum. The Parkinson's disease patients revealed relative underactivation in the left medial frontal area, right precuneus and left cerebellar hemisphere, whereas they showed relative overactivity in the left temporal cortex, right insula, left cingulate cortex and cerebellar vermis. This is the first experimental study showing that the dorsal brainstem, which corresponds to the brainstem locomotor region in experimental animals, is active during human bipedal gait. The reduced brain activity in the medial frontal motor areas is a basic abnormality in motor performance in Parkinson's disease. The underactivity in the left cerebellar hemisphere, in contrast to the overactivity in the vermis, could be associated with a loss of lateral gravity shift in parkinsonian gait. PMID- 10388792 TI - Spatial mapping of T2 and gadolinium-enhancing T1 lesion volumes in multiple sclerosis: evidence for distinct mechanisms of lesion genesis? AB - It is generally believed that most T2-weighted (T2) lesions in the central white matter of patients with multiple sclerosis begin with a variable period of T1 weighted (T1) gadolinium (Gd) enhancement and that T1 Gd-enhancing and T2 lesions represent stages of a single pathological process. Lesion probability maps can be used to test this hypothesis by providing a quantitative description of the spatial distribution of these two types of lesions across a patient population. The simplest prediction of this hypothesis would be that the spatial distributions of T1 Gd-enhancing and T2 lesions are identical. We generated T1 Gd enhancing and T2 lesion probability maps from 19 patients with relapsing remitting multiple sclerosis. There was a significantly higher probability (P = 0.001) for T2 lesions to be found in the central relative to the peripheral white matter (risk ratio 4.5), although the relative distribution of T1 Gd-enhancing lesions was not significantly different (P = 0.7) between central and peripheral white matter regions (risk ratio 0.6). Longitudinal data on the same population were used to demonstrate a similar distribution asymmetry between new T1 Gd enhancing and new T2 lesions that developed over the course of 1 year. Alternative hypotheses to explain this observation were tested. We found no spatial difference in the likelihood of development of persistent T2 lesions following T1 Gd enhancement. The relative distribution of T1 Gd-enhancing lesions was shown to be independent of the dose of Gd contrast agent and the frequency of scanning. Our findings suggest that a proportion of the periventricular T2 lesion volume may arise from mechanisms other than those associated with early breakdown of the blood-brain barrier leading to T1 Gd enhancement. PMID- 10388794 TI - Activated non-neural specific T cells open the blood-brain barrier to circulating antibodies. AB - Previous studies have shown that activated T cells can successfully cross endothelial barriers and will accumulate in tissue which contains their specific antigen. Myelin specific T cells (e.g. myelin basic protein specific) are recognized to play an important role in the induction of experimental autoimmune demyelinating disease of the CNS and have been shown to induce blood-brain barrier breakdown effectively. In this study we injected T cells reactive to a non-neural antigen (ovalbumin) systemically into Lewis rats and caused them to accumulate in the thoracic dorsal column by a prior injection of ovalbumin. Selected rats were given systemic demyelinating antibody, antimyelin oligodendrocyte antibody (anti-MOG antibody), to provide evidence of permeability changes to the blood-brain barrier. These animals were compared with control rats given systemic anti-P0 monoclonal antibody and to other rats given a direct micro injection (3 microliters) of anti-MOG antibody into the thoracic dorsal column. All animals were monitored by serial neurophysiological studies and by histological examination. Direct anti-MOG antibody injection produced a focal block in conduction at the injection site and a large circumscribed area of primary demyelination with axonal preservation within the dorsal column. An even more profound conduction block and more extensive plaque-like region of demyelination were seen in animals given antigen, activated T cells and systemic antibody. However, animals given antigen and T cells without relevant antibody did not show conduction impairment or demyelination, except when very large numbers of T cells were given; such rats developed severe irreversible axonal damage. This study demonstrates the blood-brain barrier is disrupted by activated T cells of non-neural specificity and allows large plaque-like regions of demyelination to form in the presence of circulating antimyelin antibody. The relevance of this finding to multiple sclerosis is discussed. PMID- 10388795 TI - Vestibular perception of angular velocity in normal subjects and in patients with congenital nystagmus. AB - A technique is described for the assessment of vestibular sensation. The two main goals of the study were (i) to compare the perception of angular velocity with the eye velocity output of the vestibulo-ocular reflex and (ii) to study vestibular function in patients with congenital nystagmus; this was needed since most previous studies, based on eye movement recordings, have been inconclusive. Subjects indicated their perceived angular velocity by turning by hand a wheel connected to a tachometer. The vestibular stimuli used consisted of sudden deceleration from rotation at a constant horizontal velocity of 90 degrees /s ('stopping' responses). Eye movements were recorded simultaneously with electro oculography. In normal subjects the perceived angular velocity decayed from the moment of deceleration in an exponential fashion. The mean time constant of sensation decay was approximately 16 s. Eye movement velocity decayed with a similar exponential trajectory (time constant 16 s). Congenital nystagmus patients showed markedly shortened vestibular sensation (mean time constant 7 s). The following conclusions can be drawn: (i) the similarity of the eye velocity and perceptual responses suggests that these two systems receive a vestibular signal which has been similarly processed; (ii) the time constant of the responses indicates that this vestibular signal probably originates in the same brainstem 'velocity storage' integrator; (iii) the technique described is useful for clinical assessment of vestibular function, particularly in patients with ocular motility disorders; (iv) patients with congenital nystagmus have short vestibular time constants, which is probably due to changes induced in velocity storage processing by the persistent retinal image motion present in these patients. PMID- 10388796 TI - Cerebral correlates of preserved cognitive skills in autism: a functional MRI study of embedded figures task performance. AB - When considering the cognitive abilities of people with autism, the majority of studies have explored domains in which there are deficits. However, on tests of local processing and visual search, exemplified by the Embedded Figures Task (EFT), people with autism have been reported to demonstrate superiority over normal controls. This study employed functional MRI of subjects during the performance of the EFT to test the hypothesis that normal subjects and a group with autism would activate different brain regions and that differences in the patterns of these regional activations would support distinct models of cerebral processing underlying EFT performance in the two groups. It was found that several cerebral regions were similarly activated in the two groups. However, normal controls, as well as demonstrating generally more extensive task-related activations, additionally activated prefrontal cortical areas that were not recruited in the group with autism. Conversely, subjects with autism demonstrated greater activation of ventral occipitotemporal regions. These differences in functional anatomy suggest that the cognitive strategies adopted by the two groups are different: the normal strategy invokes a greater contribution from working memory systems while the autistic group strategy depends to an abnormally large extent on visual systems for object feature analysis. This interpretation is discussed in relation to a model of autism which proposes a predisposition towards local rather than global modes of information processing. PMID- 10388797 TI - Semantic integration in reading: engagement of the right hemisphere during discourse processing. AB - We examined the brain areas involved in discourse processing by using functional MRI in 10 individuals as they read paragraphs, with or without a title, word by word for comprehension. Functional data were collected from 20 adjacent 5 mm axial slices. Discourse processing was associated with activation in inferior frontal and temporal regions of both cerebral hemispheres in the titled and untitled conditions. Moreover, there was substantially more right hemisphere activation for untitled than for the titled paragraphs. More specifically we found: (i) greater activation in the inferior temporal sulcus of both hemispheres for untitled than titled paragraphs; (ii) greater average volume of activation in response to untitled than titled paragraphs in the middle temporal sulcus of the right hemisphere and the reverse pattern in the left middle temporal sulcus. Consistent with previous studies of individuals with right hemisphere damage, we suggest that the right middle temporal regions may be especially important for integrative processes needed to achieve global coherence during discourse processing. PMID- 10388798 TI - Corticomotoneuronal synaptic connections in normal man: an electrophysiological study. AB - In order to determine the mono- or oligosynaptic character of connections between pyramidal axons and individual spinal motor neurons, we constructed peri-stimulus time histograms (PSTHs) of the firing probability of voluntarily activated single motor units (SMUs) of various upper and lower limb muscles upon slightly suprathreshold transcranial anodal electrical stimulations of the motor cortex in normal subjects. Weak anodal cortical stimuli are known to activate preferentially fast-conducting pyramidal axons directly, bypassing cell bodies and cortical interneurons. A narrow bin width (0.1 ms) was chosen to measure precisely the duration of the PSTH excitatory peak, which corresponds to the rise time of the underlying compound excitatory post-synaptic potentials (EPSP). A short duration PSTH peak indicates sharp-rising EPSPs, most commonly encountered in the case of monosynaptic connections. In flexor carpi radialis and soleus SMUs, the PSTHs of built-in responses to anodal cortical stimuli were compared with those produced by 1A afferent stimulation able to elicit a Hoffmann reflex, which is known to be largely monosynaptic. In all upper and lower limb muscles, excitable SMUs responded to anodal cortical stimuli with a highly synchronized peak of increased firing probability. In flexor carpi radialis and soleus SMUs, the mean duration of this peak was significantly narrower than that evoked by 1A afferent stimulation, indicating that monosynaptic corticomotor neuronal transmission dominates low-threshold motor units, even in proximal arm and leg muscles. In the various muscles studied, and particularly in forearm SMUs, we did not observe broad PSTH peaks against the activation of non-monosynaptic corticomotor neuronal pathways, even with near-threshold stimuli. In some triceps and forearm flexor SMUs, subthreshold anodal pulses caused significant inhibition of their voluntary firing, with a latency consistent with activation of 1A inhibitory interneurons by the descending volleys. Measurements of the maximal number of counts in the excitatory PSTH peak upon anodal cortical stimuli provide comparisons of the strength of monosynaptic inputs to various muscles which seems to be maximal for hand and finger extensor muscles, and also for deltoid. PMID- 10388799 TI - Cognitive function in primary progressive and transitional progressive multiple sclerosis: a controlled study with MRI correlates. AB - The relative rarity of primary progressive (PP) and transitional progressive (TP) multiple sclerosis has meant that little documentation of cognitive function in such patients is currently available. The aim of this study was to investigate the cognitive skills of patients with PP and TP multiple sclerosis relative to matched healthy controls, and to examine the relationship of this impairment to MRI parameters. Sixty-three patients (43 PP, 20 TP) were individually matched with healthy controls, who undertook the same cognitive tasks as the patient group. The neuropsychological assessment comprised Rao's brief repeatable battery, a reasoning test, and a measure of depression. Patients also underwent T1- and T2-weighted brain MRI. These patients were taken from a larger cohort (158 PP, 33 TP) in whom it had been demonstrated that the re were no significant differences between the mean scores of the PP and TP groups on any of the cognitive variables. The 63 patients were therefore taken as one group for comparison with the healthy controls. These patients performed significantly worse than the controls in tests of verbal memory, attention, verbal fluency and spatial reasoning. An impairment index was constructed and applied to the patient data. This correlated modestly with T2-lesion load (r = 0.45, P = 0.01), T1 hypointensity load (r = 0.45, P = 0.01) and cerebral volume (r = -0.35, P = 0.01). Thus, PP and TP multiple sclerosis patients demonstrate significant cognitive dysfunction when compared with matched healthy controls. The relationship between this impairment and MRI parameters is moderate, suggesting that cognitive dysfunction in PP and TP multiple sclerosis has a complex and multifactorial aetiology, which is not adequately explained by pathology as demonstrated on conventional MRI. PMID- 10388800 TI - Comparative analysis of gait in Parkinson's disease, cerebellar ataxia and subcortical arteriosclerotic encephalopathy. AB - Quantitative gait analysis has been used to elucidate characteristic features of neurological gait disturbances. Although a number of studies compared single patient groups with controls, there are only a few studies comparing gait parameters between patients with different neurological disorders affecting gait. In the present study, gait parameters were compared between control subjects, patients with parkinsonian gait due to idiopathic Parkinson's disease, subjects suffering from cerebellar ataxia and patients with gait disturbance due to subcortical arteriosclerotic encephalopathy. In addition to recording of baseline parameters during preferred walking velocity, subjects were required to vary velocity from very slow to very fast. Values of velocity and stride length from each subject were then used for linear regression analysis. Whereas all patient groups showed slower walking velocity and reduced step length compared with healthy controls when assessed during preferred walking, patients with ataxia and subcortical arteriosclerotic encephalopathy had, in addition, increased variability of amplitude and timing of steps. Regression analysis showed that with changing velocity, subjects with Parkinson's disease changed their stride length in the same proportion as that measured in controls. In contrast, patients with ataxia and subcortical arteriosclerotic encephalopathy had a disproportionate contribution of stride length when velocity was increased. Whereas the findings in patients with Parkinson's disease can be explained as a reduction of force gain, the observations for patients with ataxia and subcortical arteriosclerotic encephalopathy reflect an altered spatiotemporal gait strategy in order to compensate for instability. The similarity of gait disturbance in subcortical arteriosclerotic encephalopathy and cerebellar ataxia suggests common mechanisms. PMID- 10388802 TI - Right prefrontal cortex and episodic memory retrieval: a functional MRI test of the monitoring hypothesis. AB - Though the right prefrontal cortex is often activated in neuroimaging studies of episodic memory retrieval, the functional significance of this activation remains unresolved. In this functional MRI study of 12 healthy volunteers, we tested the hypothesis that one role of the right prefrontal cortex is to monitor the information retrieved from episodic memory in order to make an appropriate response. The critical comparison was between two word recognition tasks that differed only in whether correct responses did or did not require reference to the spatiotemporal context of words presented during a previous study episode. Activation in a dorsal midlateral region of the right prefrontal cortex was associated with increased contextual monitoring demands, whereas a more ventral region of the right prefrontal cortex showed retrieval-related activation that was independent of task instructions. This functional dissociation of dorsal and ventral right prefrontal regions is discussed in relation to a theoretical framework for the control of episodic memory retrieval. PMID- 10388801 TI - Abnormal cortical processing of voluntary muscle relaxation in patients with focal hand dystonia studied by movement-related potentials. AB - In order to clarify the abnormality in cortical motor preparation for voluntary muscle relaxation of the hand in patients with focal hand dystonia, Bereitschaftspotentials (BPs) preceding voluntary muscle contraction and relaxation were recorded in eight patients (three with simple writer's cramp and five with dystonic writer's cramp), and were compared with those from 10 normal subjects. Voluntary muscle relaxation: after keeping the right wrist in an extended position for > 5 s, the subject let the hand drop by voluntarily terminating muscle contraction of the wrist extensor without any associated muscle contraction. Voluntary muscle contraction: the right wrist was flexed by voluntarily contracting the wrist flexor muscle. Scalp EEGs were recorded from 11 electrodes placed over the frontal, central and parietal areas. In the control group, the BP measured at the movement onset was maximal at the left central area (C1), and distributed predominantly over the left hemisphere equally in both the contraction and relaxation tasks. In the focal hand dystonia group, BP was maximal at C1 in the contraction task, whereas, in the relaxation task, it was maximal at the midline central area (Cz) and symmetrically distributed. At the left central area, the BP amplitude in the focal hand dystonia group was diminished significantly in the relaxation task compared with the contraction task (P < 0.05). The present results demonstrate for the first time that the cortical preparatory process for voluntary muscle relaxation, or motor inhibition, is abnormal in focal hand dystonia. PMID- 10388803 TI - Spinal root and plexus hypertrophy in chronic inflammatory demyelinating polyneuropathy. AB - MRI was performed on the spinal roots, brachial and lumbar plexuses of 14 patients with chronic inflammatory demyelinating polyneuropathy (CIDP). Hypertrophy of cervical roots and brachial plexus was demonstrated in eight cases, six of whom also had hypertrophy of the lumbar plexus. Of 11 patients who received gadolinium, five of six cases with hypertrophy and one of five without hypertrophy demonstrated enhancement. All patients with hypertrophy had a relapsing-remitting course and a significantly longer disease duration. Gross onion-bulb formations were seen in a biopsy of nerve from the brachial plexus in one case with clinically evident nodular hypertrophy. We conclude that spinal root and plexus hypertrophy may be seen on MRI, particularly in cases of CIDP of long duration, and gadolinium enhancement may be present in active disease. PMID- 10388804 TI - Hardy, Weinberg and language impediments. PMID- 10388805 TI - Regulation of mRNA export by nutritional status in fission yeast. AB - We have isolated a mutation in nup184(nup184-1) that is synthetically lethal with the mRNA export defective rae1-167 mutation in Schizosaccharomyces pombe. The consequence of the synthetic lethality is a defect in mRNA export. The predicted Nup184p is similar to Nup188p of Saccharomyces cerevisiae, and a Nup184p-GFP fusion localizes to the nuclear periphery in a punctate pattern. The Deltanup184 null mutant is viable and also is synthetically lethal with rae1-167. In a rae1(+) background, both the nup184-1 and Deltanup184 mutations confer sensitivity to growth in nutrient-rich medium (YES) that is accompanied by nuclear poly(A)+ RNA accumulation. Removal of the cAMP-dependent protein kinase, Pka1p, relieved the growth and mRNA export defects of nup184 mutants when grown in nutrient-rich medium. The activation of Pka1p is necessary, but not sufficient, to cause the severe poly(A)+ RNA export defects when nup184 mutant cells are incubated in YES, suggesting nutritional status can also regulate poly(A)+ RNA export. Our results suggest that the regulation of poly(A)+ RNA export by Pka1p kinase appears to be indirect, via a translation-dependent step, but post-translationally in response to YES. PMID- 10388806 TI - Rereplication phenomenon in fission yeast requires MCM proteins and other S phase genes. AB - The fission yeast Schizosaccharomyces pombe can be induced to perform multiple rounds of DNA replication without intervening mitoses by manipulating the activity of the cyclin-dependent kinase p34(cdc2). We have examined the role in this abnormal rereplication of a large panel of genes known to be involved in normal S phase. The genes analyzed can be grouped into four classes: (1) those that have no effect on rereplication, (2) others that delay DNA accumulation, (3) several that allow a gradual increase in DNA content but not in genome equivalents, and finally, (4) mutations that completely block rereplication. The rereplication induced by overexpression of the CDK inhibitor Rum1p or depletion of the Cdc13p cyclin is essentially the same and requires the activity of two minor B-type cyclins, cig1(+) and cig2(+). In particular, the level, composition, and localization of the MCM protein complex does not alter during rereplication. Thus rereplication in fission yeast mimics the DNA synthesis of normal S phase, and the inability to rereplicate provides an excellent assay for novel S-phase mutants. PMID- 10388808 TI - Characterization of the ptr6(+) gene in fission yeast: a possible involvement of a transcriptional coactivator TAF in nucleocytoplasmic transport of mRNA. AB - Transport of mRNA from the nucleus to the cytoplasm is one of the important steps in gene expression in eukaryotic cells. To elucidate a mechanism of mRNA export, we identified a novel ptr [poly(A)+ RNA transport] mutation, ptr6, which causes accumulation of mRNA in the nucleus and inhibition of growth at the nonpermissive temperature. The ptr6(+) gene was found to encode an essential protein of 393 amino acids, which shares significant homology in amino acid sequence with yTAFII67 of budding yeast Saccharomyces cerevisiae and human hTAFII55, a subunit of the general transcription factor complex TFIID. A Ptr6p-GFP fusion protein is localized in the nucleus, suggesting that Ptr6p functions there. Northern blot analysis using probes for 10 distinct mRNAs showed that the amount of tbp+ mRNA encoding the TATA-binding protein is increased five- to sixfold, whereas amounts of others are rapidly decreased at the nonpermissive temperature in ptr6-1. ptr6 has no defects in nuclear import of an NLS-GFP fusion protein. These results suggest that Ptr6p required for mRNA transport is a Schizosaccharomyces pombe homologue of yTAFII67 and hTAFII55. This is the first report suggesting that a TAF is involved in the nucleocytoplasmic transport of mRNA in addition to the transcription of the protein-coding genes. PMID- 10388807 TI - Saccharomyces cerevisiae putative G protein, Gtr1p, which forms complexes with itself and a novel protein designated as Gtr2p, negatively regulates the Ran/Gsp1p G protein cycle through Gtr2p. AB - Prp20p and Rna1p are GDP/GTP exchanging and GTPase-activating factors of Gsp1p, respectively, and their mutations, prp20-1 and rna1-1, can both be suppressed by Saccharomyces cerevisiae gtr1-11. We found that gtr1-11 caused a single amino acid substitution in Gtr1p, forming S20L, which is a putative GDP-bound mutant protein, while Gtr1p has been reported to bind to GTP alone. Consistently, gtr1 S20N, another putative GDP-bound mutant, suppressed both prp20-1 and rna1-1. On the other hand, gtr1-Q65L, a putative GTP-bound mutant, was inhibitory to prp20-1 and rna1-1. Thus, the role that Gtr1p plays in vivo appears to depend upon the nucleotide bound to it. Our data suggested that the GTP-bound Gtr1p, but not the GDP-bound Gtr1p, interacts with itself through its C-terminal tail. S. cerevisiae possesses a novel gene, GTR2, which is homologous to GTR1. Gtr2p interacts with itself in the presence of Gtr1p. The disruption of GTR2 suppressed prp20-1 and abolished the inhibitory effect of gtr1-Q65L on prp20-1. This finding, taken together with the fact that Gtr1p-S20L is a putative, inactive GDP-bound mutant, implies that Gtr1p negatively regulates the Ran/Gsp1p GTPase cycle through Gtr2p. PMID- 10388809 TI - In vivo analysis of the domains of yeast Rvs167p suggests Rvs167p function is mediated through multiple protein interactions. AB - Morphological changes during cell division in the yeast Saccharomyces cerevisiae are controlled by cell-cycle regulators. The Pcl-Pho85p kinase complex has been implicated in the regulation of the actin cytoskeleton at least in part through Rvs167p. Rvs167p consists of three domains called BAR, GPA, and SH3. Using a two hybrid assay, we demonstrated that each region of Rvs167p participates in protein protein interactions: the BAR domain bound the BAR domain of another Rvs167p protein and that of Rvs161p, the GPA region bound Pcl2p, and the SH3 domain bound Abp1p. We identified Rvs167p as a Las17p/Bee1p-interacting protein in a two hybrid screen and showed that Las17p/Bee1p bound the SH3 domain of Rvs167p. We tested the extent to which the Rvs167p protein domains rescued phenotypes associated with deletion of RVS167: salt sensitivity, random budding, and endocytosis and sporulation defects. The BAR domain was sufficient for full or partial rescue of all rvs167 mutant phenotypes tested but not required for the sporulation defect for which the SH3 domain was also sufficient. Overexpression of Rvs167p inhibits cell growth. The BAR domain was essential for this inhibition and the SH3 domain had only a minor effect. Rvs167p may link the cell cycle regulator Pcl-Pho85p kinase and the actin cytoskeleton. We propose that Rvs167p is activated by phosphorylation in its GPA region by the Pcl-Pho85p kinase. Upon activation, Rvs167p enters a multiprotein complex, making critical contacts in its BAR domain and redundant or minor contacts with its SH3 domain. PMID- 10388810 TI - Identification and characterization of Schizosaccharomyces pombe asp1(+), a gene that interacts with mutations in the Arp2/3 complex and actin. AB - The Arp2/3 complex is an essential component of the actin cytoskeleton in yeast and is required for the movement of actin patches. In an attempt to identify proteins that interact with this complex in the fission yeast Schizosaccharomyces pombe, we sought high-copy suppressors of the S. pombe arp3-c1 mutant, and have identified one, which we have termed asp1(+). The asp1(+) open reading frame (ORF) predicts a highly conserved protein of 921 amino acids with a molecular mass of 106 kD that does not contain motifs of known function. Neither asp1(+) nor its apparent Saccharomyces cerevisiae ortholog, VIP1, are essential genes. However, disruption of asp1(+) leads to altered morphology and growth properties at elevated temperatures and defects in polarized growth. The asp1 disruption strain also is hypersensitive to Ca+ ions and to low pH conditions. Although Asp1p is not stably associated with the Arp2/3 complex nor localized in any discrete structure within the cytoplasm, the asp1 disruption mutant was synthetically lethal with mutations in components of the Arp2/3 complex, arp3-c1 and sop2-1, as well as with a mutation in actin, act1-48. Moreover, the vip1 disruption strain showed a negative genetic interaction with a las17Delta strain. We conclude that Asp1p/Vip1p is important for the function of the cortical actin cytoskeleton. PMID- 10388811 TI - Radiosensitive and mitotic recombination phenotypes of the Saccharomyces cerevisiae dun1 mutant defective in DNA damage-inducible gene expression. AB - The biological significance of DNA damage-induced gene expression in conferring resistance to DNA-damaging agents is unclear. We investigated the role of DUN1 mediated, DNA damage-inducible gene expression in conferring radiation resistance in Saccharomyces cerevisiae. The DUN1 gene was assigned to the RAD3 epistasis group by quantitating the radiation sensitivities of dun1, rad52, rad1, rad9, rad18 single and double mutants, and of the dun1 rad9 rad52 triple mutant. The dun1 and rad52 single mutants were similar in terms of UV sensitivities; however, the dun1 rad52 double mutant exhibited a synergistic decrease in UV resistance. Both spontaneous intrachromosomal and heteroallelic gene conversion events between two ade2 alleles were enhanced in dun1 mutants, compared to DUN1 strains, and elevated recombination was dependent on RAD52 but not RAD1 gene function. Spontaneous sister chromatid exchange (SCE), as monitored between truncated his3 fragments, was not enhanced in dun1 mutants, but UV-induced SCE and heteroallelic recombination were enhanced. Ionizing radiation and methyl methanesulfonate (MMS) induced DNA damage did not exhibit greater recombinogenicity in the dun1 mutant compared to the DUN1 strain. We suggest that one function of DUN1-mediated DNA damage-induced gene expression is to channel the repair of UV damage into a nonrecombinogenic repair pathway. PMID- 10388813 TI - Conservation of ARS elements and chromosomal DNA replication origins on chromosomes III of Saccharomyces cerevisiae and S. carlsbergensis. AB - DNA replication origins, specified by ARS elements in Saccharomyces cerevisiae, play an essential role in the stable transmission of chromosomes. Little is known about the evolution of ARS elements. We have isolated and characterized ARS elements from a chromosome III recovered from an alloploid Carlsberg brewing yeast that has diverged from its S. cerevisiae homeologue. The positions of seven ARS elements identified in this S. carlsbergensis chromosome are conserved: they are located in intergenic regions flanked by open reading frames homologous to those that flank seven ARS elements of the S. cerevisiae chromosome. The S. carlsbergensis ARS elements were active both in S. cerevisiae and S. monacensis, which has been proposed to be the source of the diverged genome present in brewing yeast. Moreover, their function as chromosomal replication origins correlated strongly with the activity of S. cerevisiae ARS elements, demonstrating the conservation of ARS activity and replication origin function in these two species. PMID- 10388812 TI - A general requirement for the Sin3-Rpd3 histone deacetylase complex in regulating silencing in Saccharomyces cerevisiae. AB - The Sin3-Rpd3 histone deacetylase complex, conserved between human and yeast, represses transcription when targeted by promoter-specific transcription factors. SIN3 and RPD3 also affect transcriptional silencing at the HM mating loci and at telomeres in yeast. Interestingly, however, deletion of the SIN3 and RPD3 genes enhances silencing, implying that the Sin3-Rpd3 complex functions to counteract, rather than to establish or maintain, silencing. Here we demonstrate that Sin3, Rpd3, and Sap30, a novel component of the Sin3-Rpd3 complex, affect silencing not only at the HMR and telomeric loci, but also at the rDNA locus. The effects on silencing at all three loci are dependent upon the histone deacetylase activity of Rpd3. Enhanced silencing associated with sin3Delta, rpd3Delta, and sap30Delta is differentially dependent upon Sir2 and Sir4 at the telomeric and rDNA loci and is also dependent upon the ubiquitin-conjugating enzyme Rad6 (Ubc2). We also show that the Cac3 subunit of the CAF-I chromatin assembly factor and Sin3-Rpd3 exert antagonistic effects on silencing. Strikingly, deletion of GCN5, which encodes a histone acetyltransferase, enhances silencing in a manner similar to deletion of RPD3. A model that integrates the effects of rpd3Delta, gcn5Delta, and cac3Delta on silencing is proposed. PMID- 10388814 TI - DNA sequence and functional analysis of homologous ARS elements of Saccharomyces cerevisiae and S. carlsbergensis. AB - ARS elements of Saccharomyces cerevisiae are the cis-acting sequences required for the initiation of chromosomal DNA replication. Comparisons of the DNA sequences of unrelated ARS elements from different regions of the genome have revealed no significant DNA sequence conservation. We have compared the sequences of seven pairs of homologous ARS elements from two Saccharomyces species, S. cerevisiae and S. carlsbergensis. In all but one case, the ARS308-ARS308(carl) pair, significant blocks of homology were detected. In the cases of ARS305, ARS307, and ARS309, previously identified functional elements were found to be conserved in their S. carlsbergensis homologs. Mutation of the conserved sequences in the S. carlsbergensis ARS elements revealed that the homologous sequences are required for function. These observations suggested that the sequences important for ARS function would be conserved in other ARS elements. Sequence comparisons aided in the identification of the essential matches to the ARS consensus sequence (ACS) of ARS304, ARS306, and ARS310(carl), though not of ARS310. PMID- 10388815 TI - The effect of DNA replication mutations on CAG tract stability in yeast. AB - CAG repeat tracts are unstable in yeast, leading to frequent contractions and infrequent expansions in repeat tract length. To compare CAG repeats to other simple repeats and palindromic sequences, we examined the effect of DNA replication mutations, including alleles of pol alpha, pol delta, pol epsilon, and PCNA (proliferating cell nuclear antigen), on tract stability. Among the polymerase mutations, the pol delta mutation (pol3-14) destabilizes tracts with either CAG or CTG as the lagging strand template. One pol alpha mutation, pol1-1, destabilizes the orientation with CAG as the lagging strand template, but it has little effect on the CTG orientation. In contrast, the pol1-17 mutation has no effect on either orientation. Similarly, mutations in the proofreading functions of pol delta and pol epsilon, as well as a temperature-sensitive pol epsilon mutation, pol2-18, have no effect on tract stability. Three PCNA mutations, pol30 52, pol30-79, and pol30-90, all have drastic effects on tract stability. Of the three, pol30-52 is unique in yielding small tract changes that are indicative of an impairment in mismatch repair. These results show that while CAG repeats are destabilized by many of the same mutations that destabilize other simple repeats, they also have some behaviors that are suggestive of their potential to form hairpin structures. PMID- 10388816 TI - Evidence for negative interference: clustering of crossovers close to the am locus in Neurospora crassa among am recombinants. AB - In response to a conflict between two mapping studies in the predicted orientation of the allele map with respect to the centromere, Fincham proposed that recombination events at the Neurospora am locus rarely have an associated crossover. Fincham considered that the elevated levels of crossing over between flanking markers in am recombinants resulted from negative interference, an increased probability of a nearby second event, and on this basis predicted a clustering of crossing over near am in these recombinants. In this article we reevaluate the data from three mapping studies of the am locus and report molecular evidence that shows crossovers to be clustered immediately proximal to am in am recombinants. PMID- 10388817 TI - Population structure and dynamics of Magnaporthe grisea in the Indian Himalayas. AB - The population genetics of Magnaporthe grisea, the rice blast pathogen, were analyzed in a center of rice diversity (the Uttar Pradesh hills of the Indian Himalayas) using multilocus and single-, or low-copy, DNA markers. Based on DNA fingerprinting with the multilocus probe MGR586 and single-locus probes, 157 haplotypes clustered into 56 lineages (at >/=70% MGR586 band similarity, each with unique single-locus profiles) and high diversity indices were detected among 458 isolates collected from 29 sites during 1992-1995. Most valleys sampled had distinct populations (73% of the lineages were site specific) with some containing one or a few lineages, confirming the importance of clonal propagation, and others were very diverse. Widely distributed lineages suggested that migration occurs across the region and into the Indo-Gangetic plains. Repeated sampling at one site, Matli, (170 isolates, 1992-1995) yielded 19 lineages and diversity significantly greater than that reported from similar samples from Colombia and the Philippines. Analysis of allelic associations using pairwise comparisons and multilocus variance analysis failed to reject the hypothesis of gametic phase equilibrium. The Matli population shifted from highly diverse in 1992 to almost complete dominance by one lineage in 1995. Such population dynamics are consistent with recombination followed by differential survival of clonal descendants of recombinant progeny. At another site, Ranichauri, population (n = 84) composition changed from 2 to 11 lineages over 2 yr and yielded additional evidence for equilibrium. Sexually fertile and hermaphrodite isolates of both mating types were recovered from rice in both Matli and Ranichauri. We demonstrate that Himalayan M. grisea populations are diverse and dynamic and conclude that the structure of some populations may be affected to some extent by sexual recombination. PMID- 10388818 TI - Mutations affecting symmetrical migration of distal tip cells in Caenorhabditis elegans. AB - The rotational symmetry of the Caenorhabditis elegans gonad arms is generated by the symmetrical migration of two distal tip cells (DTCs), located on the anterior and posterior ends of the gonad primordium. Mutations that cause asymmetrical migration of the two DTCs were isolated. All seven mutations were recessive and assigned to six different complementation groups. vab-3(k121) and vab-3(k143) affected anterior DTC migration more frequently than posterior, although null mutants showed no bias. The other five mutations, mig-14(k124), mig-17(k113), mig 18(k140), mig-19(k142), and mig-20(k148), affected posterior DTC migration more frequently than anterior. These observations imply that the migration of each DTC is regulated differently. mig-14 and mig-19 also affected the migration of other cells in the posterior body region. Four distinct types of DTC migration abnormalities were defined on the basis of the mutant phenotypes. vab-3; mig-14 double mutants exhibited the types of DTC migration defects seen for vab-3 single mutants. Combination of mig-17 and mig-18 or mig-19, which are characterized by the same types of posterior DTC migration defects, exhibited strong enhancement of anterior DTC migration defects, suggesting that they affect the same or parallel pathways regulating anterior DTC migration. PMID- 10388819 TI - The primary sex determination signal of Caenorhabditis elegans. AB - An X chromosome counting process determines sex in Caenorhabditis elegans. The dose of X chromosomes is translated into sexual fate by a set of X-linked genes that together control the activity of the sex-determination and dosage compensation switch gene, xol-1. The double dose of X elements in XX animals represses xol-1 expression, promoting the hermaphrodite fate, while the single dose of X elements in XO animals permits high xol-1 expression, promoting the male fate. Previous work has revealed at least four signal elements that repress xol-1 expression at two levels, transcriptional and post-transcriptional. The two molecularly characterized elements include an RNA binding protein and a nuclear hormone receptor homolog. Here we explore the roles of the two mechanisms of xol 1 repression and further investigate how the combined dose of X signal elements ensures correct, sex-specific expression of xol-1. By studying the effects of increases and decreases in X signal element dose on male and hermaphrodite fate, we demonstrate that signal elements repress xol-1 cumulatively, such that full repression of xol-1 in XX animals results from the combined effect of individual elements. Complete transformation from the hermaphrodite to the male fate requires a decrease in the dose of all four elements, from two copies to one. We show that both mechanisms of xol-1 repression are essential and act synergistically to keep xol-1 levels low in XX animals. However, increasing repression by one mechanism can compensate for loss of the other, demonstrating that each mechanism can exert significant xol-1 repression on its own. Finally, we present evidence suggesting that xol-1 activity can be set at intermediate levels in response to an intermediate X signal. PMID- 10388820 TI - Selective sweep at the Drosophila melanogaster Suppressor of Hairless locus and its association with the In(2L)t inversion polymorphism. AB - The hitchhiking model of population genetics predicts that an allele favored by Darwinian selection can replace haplotypes from the same locus previously established at a neutral mutation-drift equilibrium. This process, known as "selective sweep," was studied by comparing molecular variation between the polymorphic In(2L)t inversion and the standard chromosome. Sequence variation was recorded at the Suppressor of Hairless (Su[H]) gene in an African population of Drosophila melanogaster. We found 47 nucleotide polymorphisms among 20 sequences of 1.2 kb. Neutrality tests were nonsignificant at the nucleotide level. However, these sites were strongly associated, because 290 out of 741 observed pairwise combinations between them were in significant linkage disequilibrium. We found only seven haplotypes, two occurring in the 9 In(2L)t chromosomes, and five in the 11 standard chromosomes, with no shared haplotype. Two haplotypes, one in each chromosome arrangement, made up two-thirds of the sample. This low haplotype diversity departed from neutrality in a haplotype test. This pattern supports a selective sweep hypothesis for the Su(H) chromosome region. PMID- 10388821 TI - Molecular cloning and tissue-specific expression of the mutator2 gene (mu2) in Drosophila melanogaster. AB - We present here the molecular cloning and characterization of the mutator2 (mu2) gene of Drosophila melanogaster together with further genetic analyses of its mutant phenotype. mu2 functions in oogenesis during meiotic recombination, during repair of radiation damage in mature oocytes, and in proliferating somatic cells, where mu2 mutations cause an increase in somatic recombination. Our data show that mu2 represents a novel component in the processing of double strand breaks (DSBs) in female meiosis. mu2 does not code for a DNA repair enzyme because mu2 mutants are not hypersensitive to DSB-inducing agents. We have mapped and cloned the mu2 gene and rescued the mu2 phenotype by germ-line transformation with genomic DNA fragments containing the mu2 gene. Sequencing its cDNA demonstrates that mu2 encodes a novel 139-kD protein, which is highly basic in the carboxy half and carries three nuclear localization signals and a helix-loop-helix domain. Consistent with the sex-specific mutant phenotype, the gene is expressed in ovaries but not in testes. During oogenesis its RNA is rapidly transported from the nurse cells into the oocyte where it accumulates specifically at the anterior margin. Expression is also prominent in diploid proliferating cells of larval somatic tissues. Our genetic and molecular data are consistent with the model that mu2 encodes a structural component of the oocyte nucleus. The MU2 protein may be involved in controlling chromatin structure and thus may influence the processing of DNA DSBs. PMID- 10388822 TI - I-SceI endonuclease, a new tool for studying DNA double-strand break repair mechanisms in Drosophila. AB - As a step toward the development of a homologous recombination system in Drosophila, we have developed a methodology to target double-strand breaks (DSBs) to a specific position in the Drosophila genome. This method uses the mitochondrial endonuclease I-SceI that recognizes and cuts an 18-bp restriction site. We find that >6% of the progeny derived from males that carry a marker gene bordered by two I-SceI sites and that express I-SceI in their germ line lose the marker gene. Southern blot analysis and sequencing of the regions surrounding the I-SceI sites revealed that in the majority of the cases, the introduction of DSBs at the I-SceI sites resulted in the complete deletion of the marker gene; the other events were associated with partial deletion of the marker gene. We discuss a number of applications for this novel technique, in particular its use to study DSB repair mechanisms. PMID- 10388823 TI - The lawc gene is a new member of the trithorax-group that affects the function of the gypsy insulator of Drosophila. AB - Mutations in the lawc gene result in a pleiotropic phenotype that includes homeotic transformation of the arista into leg. lawc mutations enhance the phenotype of trx-G mutations and suppress the phenotype of Pc mutations. Mutations in lawc affect homeotic gene transcription, causing ectopic expression of Antennapedia in the eye-antenna imaginal disc. These results suggest that lawc is a new member of the trithorax family. The lawc gene behaves as an enhancer of position-effect variegation and interacts genetically with mod(mdg4), which is a component of the gypsy insulator. In addition, mutations in the lawc gene cause alterations in the punctated distribution of mod(mdg4) protein within the nucleus. These results suggest that the lawc protein is involved in regulating the higher-order organization of chromatin. PMID- 10388824 TI - Clustered microsatellite mutations in the pipefish Syngnathus typhle. AB - Clustered mutations are copies of a mutant allele that enter a population's gene pool together due to replication from a premeiotic germline mutation and distribution to multiple successful gametes of an individual. Although the phenomenon has been studied in Drosophila and noted in a few other species, the topic has received scant attention despite claims of being of major importance to population genetics theory. Here we capitalize upon the reproductive biology of male-pregnant pipefishes to document the occurrence of clustered microsatellite mutations and to estimate their rates and patterns from family data. Among a total of 3195 embryos genetically screened from 110 families, 40% of the 35 detected de novo mutant alleles resided in documented mutational clusters. Most of the microsatellite mutations appeared to involve small-integer changes in repeat copy number, and they arose in approximately equal frequency in paternal and maternal germlines. These findings extend observations on clustered mutations to another organismal group and motivate a broader critique of the mutation cluster phenomenon. They also carry implications for the evolution of microsatellites with respect to mutational models and homoplasy among alleles. PMID- 10388826 TI - Estimation of past demographic parameters from the distribution of pairwise differences when the mutation rates vary among sites: application to human mitochondrial DNA. AB - Distributions of pairwise differences often called "mismatch distributions" have been extensively used to estimate the demographic parameters of past population expansions. However, these estimations relied on the assumption that all mutations occurring in the ancestry of a pair of genes lead to observable differences (the infinite-sites model). This mutation model may not be very realistic, especially in the case of the control region of mitochondrial DNA, where this methodology has been mostly applied. In this article, we show how to infer past demographic parameters by explicitly taking into account a finite sites model with heterogeneity of mutation rates. We also propose an alternative way to derive confidence intervals around the estimated parameters, based on a bootstrap approach. By checking the validity of these confidence intervals by simulations, we find that only those associated with the timing of the expansion are approximately correctly estimated, while those around the population sizes are overly large. We also propose a test of the validity of the estimated demographic expansion scenario, whose proper behavior is verified by simulation. We illustrate our method with human mitochondrial DNA, where estimates of expansion times are found to be 10-20% larger when taking into account heterogeneity of mutation rates than under the infinite-sites model. PMID- 10388825 TI - Influence of sex, smoking and age on human hprt mutation frequencies and spectra. AB - Examination of the literature for hprt mutant frequencies from peripheral T cells yielded data from 1194 human subjects. Relationships between mutant frequency, age, sex, and smoking were examined, and the kinetics were described. Mutant frequency increases rapidly with age until about age 15. Afterward, the rate of increase falls such that after age 53, the hprt mutant frequency is largely stabilized. Sex had no effect on mutant frequency. Cigarette smoking increased mean mutant frequency compared to nonsmokers, but did not alter age vs. mutant frequency relationships. An hprt in vivo mutant database containing 795 human hprt mutants from 342 individuals was prepared. No difference in mutational spectra was observed comparing smokers to nonsmokers, confirming previous reports. Sex affected the frequency of deletions (>1 bp) that are recovered more than twice as frequently in females (P = 0. 008) compared to males. There is no indication of a significant shift in mutational spectra with age for individuals older than 19 yr, with the exception of A:T --> C:G transversions. These events are recovered more frequently in older individuals. PMID- 10388827 TI - Spatial and temporal distribution of the neutral polymorphisms in the last ZFX intron: analysis of the haplotype structure and genealogy. AB - With 10 segregating sites (simple nucleotide polymorphisms) in the last intron (1089 bp) of the ZFX gene we have observed 11 haplotypes in 336 chromosomes representing a worldwide array of 15 human populations. Two haplotypes representing 77% of all chromosomes were distributed almost evenly among four continents. Five of the remaining haplotypes were detected in Africa and 4 others were restricted to Eurasia and the Americas. Using the information about the ancestral state of the segregating positions (inferred from human-great ape comparisons), we applied coalescent analysis to estimate the age of the polymorphisms and the resulting haplotypes. The oldest haplotype, with the ancestral alleles at all the sites, was observed at low frequency only in two groups of African origin. Its estimated age of 740 to 1100 kyr corresponded to the time to the most recent common ancestor. The two most frequent worldwide distributed haplotypes were estimated at 550 to 840 and 260 to 400 kyr, respectively, while the age of the continentally restricted polymorphisms was 120 to 180 kyr and smaller. Comparison of spatial and temporal distribution of the ZFX haplotypes suggests that modern humans diverged from the common ancestral stock in the Middle Paleolithic era. Subsequent range expansion prevented substantial gene flow among continents, separating African groups from populations that colonized Eurasia and the New World. PMID- 10388828 TI - Pattern of nucleotide substitution and rate heterogeneity in the hypervariable regions I and II of human mtDNA. AB - This study provides a comprehensive survey of the complex pattern of nucleotide substitution in the control region of human mtDNA, which is of central importance to the studies of human evolution. A total of 1229 different hypervariable region I (HVRI) and 385 different hypervariable region II (HVRII) sequences were analyzed using a complex substitution model. Moreover, we suggest a new method to assign relative rates to each site in the sequence. Estimates are based on maximum-likelihood methods applied to randomly selected subsets of sequences. Our results indicate that the rate of substitution in HVRI is approximately twice as high as in HVRII and that this difference is mainly due to a higher frequency of pyrimidine transitions in HVRI. However, rate heterogeneity is more pronounced in HVRII. PMID- 10388829 TI - Homeologous plastid DNA transformation in tobacco is mediated by multiple recombination events. AB - Efficient plastid transformation has been achieved in Nicotiana tabacum using cloned plastid DNA of Solanum nigrum carrying mutations conferring spectinomycin and streptomycin resistance. The use of the incompletely homologous (homeologous) Solanum plastid DNA as donor resulted in a Nicotiana plastid transformation frequency comparable with that of other experiments where completely homologous plastid DNA was introduced. Physical mapping and nucleotide sequence analysis of the targeted plastid DNA region in the transformants demonstrated efficient site specific integration of the 7.8-kb Solanum plastid DNA and the exclusion of the vector DNA. The integration of the cloned Solanum plastid DNA into the Nicotiana plastid genome involved multiple recombination events as revealed by the presence of discontinuous tracts of Solanum-specific sequences that were interspersed between Nicotiana-specific markers. Marked position effects resulted in very frequent cointegration of the nonselected peripheral donor markers located adjacent to the vector DNA. Data presented here on the efficiency and features of homeologous plastid DNA recombination are consistent with the existence of an active RecA-mediated, but a diminished mismatch, recombination/repair system in higher-plant plastids. PMID- 10388830 TI - A molecular description of mutations affecting the pollen component of the Nicotiana alata S locus. AB - Mutations affecting the self-incompatibility response of Nicotiana alata were generated by irradiation. Mutants in the M1 generation were selected on the basis of pollen tube growth through an otherwise incompatible pistil. Twelve of the 18 M1 plants obtained from the mutagenesis screen were self-compatible. Eleven self compatible plants had mutations affecting only the pollen function of the S locus (pollen-part mutants). The remaining self-compatible plant had a mutation affecting only the style function of the S locus (style-part mutant). Cytological examination of the pollen-part mutant plants revealed that 8 had an extra chromosome (2n + 1) and 3 did not. The pollen-part mutation in 7 M1 plants was followed in a series of crosses. DNA blot analysis using probes for S-RNase genes (encoding the style function of the S locus) indicated that the pollen-part mutation was associated with an extra S allele in 4 M1 plants. In 3 of these plants, the extra S allele was located on the additional chromosome. There was no evidence of an extra S allele in the 3 remaining M1 plants. The breakdown of self incompatibility in plants with an extra S allele is discussed with reference to current models of the molecular basis of self-incompatibility. PMID- 10388831 TI - A maize map standard with sequenced core markers, grass genome reference points and 932 expressed sequence tagged sites (ESTs) in a 1736-locus map. AB - We have constructed a 1736-locus maize genome map containing1156 loci probed by cDNAs, 545 probed by random genomic clones, 16 by simple sequence repeats (SSRs), 14 by isozymes, and 5 by anonymous clones. Sequence information is available for 56% of the loci with 66% of the sequenced loci assigned functions. A total of 596 new ESTs were mapped from a B73 library of 5-wk-old shoots. The map contains 237 loci probed by barley, oat, wheat, rice, or tripsacum clones, which serve as grass genome reference points in comparisons between maize and other grass maps. Ninety core markers selected for low copy number, high polymorphism, and even spacing along the chromosome delineate the 100 bins on the map. The average bin size is 17 cM. Use of bin assignments enables comparison among different maize mapping populations and experiments including those involving cytogenetic stocks, mutants, or quantitative trait loci. Integration of nonmaize markers in the map extends the resources available for gene discovery beyond the boundaries of maize mapping information into the expanse of map, sequence, and phenotype information from other grass species. This map provides a foundation for numerous basic and applied investigations including studies of gene organization, gene and genome evolution, targeted cloning, and dissection of complex traits. PMID- 10388832 TI - Double-strand break-induced recombination between ectopic homologous sequences in somatic plant cells. AB - Homologous recombination between ectopic sites is rare in higher eukaryotes. To test whether double-strand breaks (DSBs) can induce ectopic recombination, transgenic tobacco plants harboring two unlinked, nonfunctional homologous parts of a kanamycin resistance gene were produced. To induce homologous recombination between the recipient locus (containing an I-SceI site within homologous sequences) and the donor locus, the rare cutting restriction enzyme I-SceI was transiently expressed via Agrobacterium in these plants. Whereas without I-SceI expression no recombination events were detectable, four independent recombinants could be isolated after transient I-SceI expression, corresponding to approximately one event in 10(5) transformations. After regeneration, the F1 generation of all recombinants showed Mendelian segregation of kanamycin resistance. Molecular analysis of the recombinants revealed that the resistance gene was indeed restored via homologous recombination. Three different kinds of reaction products could be identified. In one recombinant a classical gene conversion without exchange of flanking markers occurred. In the three other cases homologous sequences were transferred only to one end of the break. Whereas in three cases the ectopic donor sequence remained unchanged, in one case rearrangements were found in recipient and donor loci. Thus, ectopic homologous recombination, which seems to be a minor repair pathway for DSBs in plants, is described best by recombination models that postulate independent roles for the break ends during the repair process. PMID- 10388833 TI - Genome mapping in capsicum and the evolution of genome structure in the solanaceae. AB - We have created a genetic map of Capsicum (pepper) from an interspecific F2 population consisting of 11 large (76.2-192.3 cM) and 2 small (19.1 and 12.5 cM) linkage groups that cover a total of 1245.7 cM. Many of the markers are tomato probes that were chosen to cover the tomato genome, allowing comparison of this pepper map to the genetic map of tomato. Hybridization of all tomato-derived probes included in this study to positions throughout the pepper map suggests that no major losses have occurred during the divergence of these genomes. Comparison of the pepper and tomato genetic maps showed that 18 homeologous linkage blocks cover 98.1% of the tomato genome and 95.0% of the pepper genome. Through these maps and the potato map, we determined the number and types of rearrangements that differentiate these species and reconstructed a hypothetical progenitor genome. We conclude there have been 30 breaks as part of 5 translocations, 10 paracentric inversions, 2 pericentric inversions, and 4 disassociations or associations of genomic regions that differentiate tomato, potato, and pepper, as well as an additional reciprocal translocation, nonreciprocal translocation, and a duplication or deletion that differentiate the two pepper mapping parents. PMID- 10388835 TI - Probability of identity by descent in metapopulations. AB - Equilibrium probabilities of identity by descent (IBD), for pairs of genes within individuals, for genes between individuals within subpopulations, and for genes between subpopulations are calculated in metapopulation models with fixed or varying colony sizes. A continuous-time analog to the Moran model was used in either case. For fixed-colony size both propagule and migrant pool models were considered. The varying population size model is based on a birth-death immigration (BDI) process, to which migration between colonies is added. Wright's F statistics are calculated and compared to previous results. Adding between island migration to the BDI model can have an important effect on the equilibrium probabilities of IBD and on Wright's index. PMID- 10388834 TI - Multiple interval mapping for quantitative trait loci. AB - A new statistical method for mapping quantitative trait loci (QTL), called multiple interval mapping (MIM), is presented. It uses multiple marker intervals simultaneously to fit multiple putative QTL directly in the model for mapping QTL. The MIM model is based on Cockerham's model for interpreting genetic parameters and the method of maximum likelihood for estimating genetic parameters. With the MIM approach, the precision and power of QTL mapping could be improved. Also, epistasis between QTL, genotypic values of individuals, and heritabilities of quantitative traits can be readily estimated and analyzed. Using the MIM model, a stepwise selection procedure with likelihood ratio test statistic as a criterion is proposed to identify QTL. This MIM method was applied to a mapping data set of radiata pine on three traits: brown cone number, tree diameter, and branch quality scores. Based on the MIM result, seven, six, and five QTL were detected for the three traits, respectively. The detected QTL individually contributed from approximately 1 to 27% of the total genetic variation. Significant epistasis between four pairs of QTL in two traits was detected, and the four pairs of QTL contributed approximately 10.38 and 14.14% of the total genetic variation. The asymptotic variances of QTL positions and effects were also provided to construct the confidence intervals. The estimated heritabilities were 0.5606, 0.5226, and 0. 3630 for the three traits, respectively. With the estimated QTL effects and positions, the best strategy of marker-assisted selection for trait improvement for a specific purpose and requirement can be explored. The MIM FORTRAN program is available on the worldwide web (http://www.stat.sinica.edu.tw/chkao/). PMID- 10388836 TI - Tales from the front lines: the creative essay as a tool for teaching genetics. AB - In contrast to the more typical mock grant proposals or literature reviews, we describe the use of the creative essay as a novel tool for teaching human genetics at the college level. This method has worked well for both nonmajor and advanced courses for biology majors. The 10- to 15-page essay is written in storylike form and represents a student's response to the choice of 6-8 scenarios describing human beings coping with various genetic dilemmas. We have found this tool to be invaluable both in developing students' ability to express genetic concepts in lay terms and in promoting student awareness of genetic issues outside of the classroom. Examples from student essays are presented to illustrate these points, and guidelines are suggested regarding instructor expectations of student creativity and scientific accuracy. Methods of grading this assignment are also discussed. PMID- 10388838 TI - Comment on a paper by Facchiano et al. (1998). Evaluation of relationship between helix stability and protein thermostability. PMID- 10388839 TI - Reply to comment PMID- 10388840 TI - Crystal structure of human serum albumin at 2.5 A resolution. AB - A new triclinic crystal form of human serum albumin (HSA), derived either from pool plasma (pHSA) or from a Pichia pastoris expression system (rHSA), was obtained from polyethylene glycol 4000 solution. Three-dimensional structures of pHSA and rHSA were determined at 2.5 A resolution from the new triclinic crystal form by molecular replacement, using atomic coordinates derived from a multiple isomorphous replacement work with a known tetragonal crystal form. The structures of pHSA and rHSA are virtually identical, with an r.m. s. deviation of 0.24 A for all Calpha atoms. The two HSA molecules involved in the asymmetric unit are related by a strict local twofold symmetry such that the Calpha atoms of the two molecules can be superimposed with an r.m.s. deviation of 0.28 A in pHSA. Cys34 is the only cysteine with a free sulfhydryl group which does not participate in a disulfide linkage with any external ligand. Domains II and III both have a pocket formed mostly of hydrophobic and positively charged residues and in which a very wide range of compounds may be accommodated. Three tentative binding sites for long-chain fatty acids, each with different surroundings, are located at the surface of each domain. PMID- 10388841 TI - Secondary structures without backbone: an analysis of backbone mimicry by polar side chains in protein structures. AB - Backbone mimicry by the formation of closed-loop C7, C10 and C13 (mimics of gamma , beta- and alpha-turns) conformations through side chain-main chain hydrogen bonds by polar groups is a frequent observation in protein structures. A data set of 250 non-homologous and high-resolution protein crystal structures was used to analyze these conformations for their characteristic features. Seven out of the nine polar residues (Ser, Thr, Asn, Asp, Gln, Glu and His) have hydrogen bonding groups in their side chains which can participate in such mimicry and as many as 15% of all these polar residues engage in such conformations. The distributions of dihedral angles of these mimics indicate that only certain combinations of the dihedral angles involved aid the formation of these mimics. The observed examples were categorized into various classes based on these combinations, resulting in well defined motifs. Asn and Asp residues show a very high capability to perform such backbone secondary structural mimicry. The most highly mimicked backbone structure is of the C10 conformation by the Asx residues. The mimics formed by His, Ser, Thr and Glx residues are also discussed. The role of such conformations in initiating the formation of regular secondary structures during the course of protein folding seems significant. PMID- 10388842 TI - A method for computational combinatorial peptide design of inhibitors of Ras protein. AB - A computational combinatorial approach is proposed for the design of a peptide inhibitor of Ras protein. The procedure involves three steps. First, a 'Multiple Copy Simultaneous Search' identifies the location of specific functional groups on the Ras surface. This search method allowed us to identify an important binding surface consisting of two beta strands (residues 5-8 and 52-56), in addition to the well known Ras effector loop and switch II region. The two beta strands had not previously been reported to be involved in Ras-Raf interaction. Second, after constructing the peptide inhibitor chain based on the location of N methylacetamide (NMA) minima, functional groups are selected and connected to the main chain Calpha atom. This step generates a number of possible peptides with different sequences on the Ras surface. Third, potential inhibitors are designed based on a sequence alignment of the peptides generated in the second step. This computational approach reproduces the conserved pattern of hydrophobic, hydrophilic and charged amino acids identified from the Ras effectors. The advantages and limitations of this approach are discussed. PMID- 10388843 TI - Molecular mechanics analysis of drug-resistant mutants of HIV protease. AB - Drug-resistant mutants of HIV-1 protease limit the long-term effectiveness of current anti-viral therapy. In order to study drug resistance, the wild-type HIV 1 protease and the mutants R8Q, V32I, M46I, V82A, V82I, V82F, I84V, V32I/I84V and M46I/I84V were modeled with the inhibitors saquinavir and indinavir using the program AMMP. A new screen term was introduced to reproduce more correctly the electron distribution of atoms. The atomic partial charge was represented as a delocalized charge distribution instead of a point charge. The calculated protease-saquinavir interaction energies showed the highly significant correlation of 0.79 with free energy differences derived from the measured inhibition constants for all 10 models. Three different protonation states of indinavir were evaluated. The best indinavir model included a sulfate and gave a correlation coefficient of 0.68 between the calculated interaction energies and free energies from inhibition constants for nine models. The exception was R8Q with indinavir, probably due to differences in the solvation energy. No significant correlation was found using the standard molecular mechanics terms. The incorporation of the new screen correction resulted in better prediction of the effects of inhibitors on resistant protease variants and has potential for selecting more effective inhibitors for resistant virus. PMID- 10388844 TI - Structural characterization by computer experiments of the lipid-free LDL receptor-binding domain of apolipoprotein E. AB - The structure and dynamics of the lipid-free LDL-receptor-binding domain of apolipoprotein E (apoE-RBD) has been investigated by Molecular Dynamics Simulations. ApoE-RBD in its monomeric lipid-free form is a singular four-helix bundle made up of four elongated amphipathic helices. Analysis of one 1.5 ns molecular dynamics trajectory of apoE-RBD performed in water indicates that the lipid-free domain adopts a structure that exhibits characteristics found in native proteins: it has very stable helices and presents a compact structure. Yet its interior exhibits a larger number of transient atomic-size cavities relative to that found in other proteins of similar size and its apolar side chains are more mobile. The latter features distinguish the elongated four-helix bundle as a slightly disordered structure, which shows a structural likeness with some de novo designed four-helix bundle proteins and shares with the latter a leucine rich residue composition. We anticipate that these unique properties compared with other native helix bundles may be related to the postulated ability of apoE RBD to undergo an opening of its bundle upon interaction with phospholipids. The distribution of empty cavities computed along the trajectory in the interface regions between the different pairs of helices reveals that the tertiary contacts in one of the interfaces are weaker suggesting that this particular interface could be more easily ruptured upon lipid association. PMID- 10388845 TI - Direct energy transfer to study the 3D structure of non-native proteins: AGH complex in molten globule state of apomyoglobin. AB - The direct energy transfer technique was modified and applied to probe the relative localization of apomyoglobin A-, G- and H-helixes, which are partly protected from deuterium exchange in the equilibrium molten globule state and in the molten globule-like kinetic intermediate. The non-radiative transfer of tryptophan electronic energy to 3-nitrotyrosine was studied in different conformational states of apomyoglobin (native, molten globule, unfolded) and interpreted in terms of average distances between groups of the protein chain. The experimental data show that the distance between the middle of A-helix and the N-terminus of G-helix as well as the distance between the middle of the A helix and the C-terminus of the H-helix in the molten globule state are close to those in the native state. This is a strong argument in favor of similarity of the overall architecture of the molten globule and native states. PMID- 10388846 TI - A general method for relieving substrate inhibition in lactate dehydrogenases. AB - The mutation S163L in human heart lactate dehydrogenase removes substrate inhibition while only modestly reducing the turnover rate for pyruvate. Since this is the third enzyme to show this behaviour, we suggest that the S163L mutation is a general method for the removal of substrate inhibition in L-LDH enzymes. Engineering such enzymatic properties has clear industrial applications in the use of these enzymes to produce enantiomerically pure alpha-hydroxy acids. The mutation leads to two principal effects. (1) Substrate inhibition is caused by the formation of a covalent adduct between pyruvate and the oxidized form of the cofactor. The inability of S163L mutants to catalyse the formation of this inhibitory adduct is demonstrated. However, NMR experiments show that the orientation of the nicotinamide ring in the mutant NAD+ binary complex is not perturbed. (2) The mutation also leads to a large increase in the KM for pyruvate. The kinetic and binding properties of S163L LDH mutants are accounted for by a mechanism which invokes a non-productive, bound form of the cofactor. Molecular modelling suggests a structure for this non-productive enzyme-NADH complex. PMID- 10388847 TI - Catalytic role of the active site histidine of porcine pancreatic phospholipase A2 probed by the variants H48Q, H48N and H48K. AB - The catalytic contribution of His48 in the active site of porcine pancreatic phospholipase A2 was examined using site-directed mutagenesis. Replacement of His48 by lysine (H48K) gives rise to a protein having a distorted lipid binding pocket. Activity of this variant drops below the detection limit which is 10(7) fold lower than that of the wild-type enzyme. On the other hand, the presence of glutamine (H48Q) or asparagine (H48N) at this position does not affect the structural integrity of the enzyme as can be derived from the preserved lipid binding properties of these variants. However, the substitutions H48Q and H48N strongly reduce the turnover number, i.e. by a factor of 10(5). Residual activity is totally lost after addition of a competitive inhibitor. We conclude that proper lipid binding on its own accelerates ester bond hydrolysis by a factor of 10(2). With the selected variants, we were also able to dissect the contribution of the hydrogen bond between Asp99 and His48 on conformational stability, being 5.2 kJ/mol. Another hydrogen bond with His48 is formed when the competitive inhibitor (R)-2-dodecanoylamino-hexanol-1-phosphoglycol interacts with the enzyme. Its contribution to binding of the inhibitor in the presence of an interface was found to be 5.7 kJ/mol. PMID- 10388848 TI - Engineering a soluble extracellular erythropoietin receptor (EPObp) in Pichia pastoris to eliminate microheterogeneity, and its complex with erythropoietin. AB - The extracellular ligand-binding domain (EPObp) of the human EPO receptor (EPOR) was expressed both in CHO (Chinese Hamster Ovary) cells and in Pichia pastoris. The CHO and yeast expressed receptors showed identical affinity for EPO binding. Expression levels in P. pastoris were significantly higher, favoring its use as an expression and scale-up production system. Incubation of EPO with a fourfold molar excess of receptor at high protein concentrations yielded stable EPO-EPObp complexes. Quantification of EPO and EPObp in the complex yielded a molar ratio of one EPO molecule to two receptor molecules. Residues that are responsible for EPOR glycosylation and isomerization in Pichia were identified and eliminated by site-specific mutagenesis. A thiol modification was identified and a method was developed to remove the modified species from EPObp. EPObp was complexed with erythropoietin (EPO) and purified. The complex crystallized in two crystal forms that diffracted to 2.8 and 1.9 A respectively. (Form 1 and form 2 crystals were independently obtained at AxyS Pharmaceuticals, Inc. and Amgen, Inc. respectively.) Both contained one complex per asymmetric unit with a stoichiometry of two EPObps to one EPO. PMID- 10388849 TI - Recombinant anti-polyamine antibodies: identification of a conserved binding site motif. AB - Polyamines are small linear polycations found ubiquitously in eukaryotic cells. They are involved in nucleic acid and protein synthesis and rises in cellular polyamine levels have been correlated with cell proliferation. Antibodies to these molecules have potential as prognostic indicators of disease conditions and indicators of treatment efficacy. Antipolyamine monoclonal antibodies of differing but defined specificities have been generated in our laboratory using polyamine ovalbumin conjugates as immunogens. These antibodies show small but significant cross reactivities with other polyamine species; IAG-1 cross reacts with spermidine (8%), JAC-1 with spermine (6%) and JSJ-1 with both putrescine (11%) and spermine (6%). We have rescued and sequenced the heavy and light chain variable regions of all three of these antibodies. While the light chains of two antibodies, IAG-1 and JSJ-1, were 93% homologous at the amino acid level, none of the heavy chains displayed any significant sequence homology. However, computer generated models of all three antibody binding sites revealed a three dimensionally conserved polyamine binding site motif. The polyamine appears to bind into a negatively charged cleft lined with acidic and polar residues. The cleft is partially or completely closed at one end and the specificity of the interaction is determined by placement of acidic residues in the cleft. Aromatic residues contribute to polyamine binding interacting with the carbon backbone. The polyamine-binding motif we have identified is very similar to that observed in the crystal structure of PotD, the primary receptor of the polyamine transport system in Escherichia coli. PMID- 10388850 TI - Human tissue responses to metal stents. AB - Metal stents have become an important addition to therapeutic strategies available for incurable gastrointestinal malignancies. The responses of human tissues to the presence of metal stents are important for several reasons. The first is to understand the mechanism by which stents are anchored in the stenosis, as this will prevent migration of stents. The second is to develop new designs of stents that would be removable. The third reason is to prevent complications of stents, such as benign hypertrophy at the proximal and distal ends of the stent, and to understand the mechanism of serious complications, such as migration through the esophageal wall or aortoesophageal fistula. In this article, the authors discuss the state of the current knowledge in these three areas. PMID- 10388852 TI - The wallstent endoprosthesis. AB - The Wallstent endoprosthesis is the most widely used self-expanding metal stent for treating digestive disease worldwide. This article presents detailed metallurgical and physical information on the Wallstent endoprosthesis, and addresses implications for its clinical use. Detailed product information provides technical insight into the implantable stent and associated delivery systems. Current worldwide product indications and configurations are discussed, as well as insight into new products in clinical evaluation. PMID- 10388851 TI - The Z-stent. AB - Z-stents were originally designed to relieve upper or lower vena cava obstructions. In time, they were adapted to be somewhat flexible, yet keep their position. This article presents information on the technology behind the Z-stent, placement of the Wilson-Cook Z-stent, and future manufacturer developments. PMID- 10388853 TI - The ultraflex esophageal and diamond biliary stents. AB - This article begins with an overview of the history and development of stents. The material properties unique to the Ultraflex esophageal stent and the Diamond biliary stent along with their clinical benefits are presented in detail. The author also provides detailed product information, recommendations, and contraindications for the use of both stents. PMID- 10388854 TI - Z-stent for malignant esophageal obstruction. AB - The Z-stent was one of the first self-expanding metal prostheses used for palliation of malignant esophageal obstruction and respiratory esophageal fistula. Its placement has proved to be effective and relatively safe. This article reviews the evolution of the Z-stent; its multiple designs, placement technique, efficacy, complications, and assets and limitations. PMID- 10388855 TI - The wallstent I and II for malignant esophageal obstruction. AB - The Wallstent I and II esophageal endoprostheses are effective in the palliation of malignant esophageal obstruction and the closure of digestive-respiratory fistulas. The author reviews the evolution of the Wallstent esophageal endoprosthesis and techniques for successful insertion, and summarizes the published literature with regard to clinical outcomes of the European and American Wallstent designs. PMID- 10388856 TI - The ultraflex stents for malignant esophageal obstruction. AB - Ultraflex esophageal stents have contributed to the tremendous success of self expanding metal stents (SEMS) in the treatment of esophageal cancer because they are easy and safe to insert. With an eye to improving clinical outcome, the Ultraflex stent design has been in a state of constant evolution since its introduction. However, as with other SEMS, a high reintervention rate remains a challenging problem. PMID- 10388857 TI - The esophacoil stent for malignant esophageal obstruction. AB - Esophageal cancer is the seventh most common cause of cancer-related deaths in men in the United States. For a disease in which 60% of patients are incurable at presentation, palliative therapy is the main treatment option. Palliative measures include surgery, radiation therapy, chemotherapy, photodynamic therapy, endoscopic dilation, endoscopic ablation, and the use of endoscopic prostheses. This article reviews the characteristics, technique, efficacy, and complications of the Esophacoil stent for malignant esophageal strictures. PMID- 10388859 TI - Expandable metal stents for benign esophageal obstruction. AB - Extensive experience with expandable metal stents for benign esophageal obstruction is limited. A review of the literature demonstrates a high incidence of complications varying from stent migration to stent-induced trauma leading to fistulization. The most common complication (41%) is that of stent-induced stenosis caused by granulation tissue and fibrosis. Currently, the authors do not recommend the routine use of expandable metal stents for benign obstruction. Each case must be assessed on its own merits and risks. The solution in the future may well have to be new stent configuration and esophagus-friendly materials. PMID- 10388858 TI - The radiologist's view of expandable metallic stents for malignant esophageal obstruction. AB - Interventional radiologists place esophageal stents with great accuracy and low complication rates. The results of radiologic placement are similar to endoscopic stent insertion. The advantages of radiologic stent insertion are the ability to traverse very small strictures that cannot be crossed endoscopically and the ability to visualize and treat small fistulae and perforations at the time of stenting. PMID- 10388860 TI - Expandable metal stents for gastric-outlet, duodenal, and small intestinal obstruction. AB - The treatment of patients who have malignant gastric-outlet, duodenal and small intestinal obstructions is difficult. The morbidity and mortality of palliative surgery in these patients is significant. It is not uncommon for patients to be treated with supportive therapy only, which unfortunately, neither relieves the severe nausea and vomiting, nor allows adequate food intake. Over the past few years, a number of studies have reported the safety and efficacy of self expanding metal stents used to palliate malignant upper gastrointestinal obstruction. In this article, the authors focus on the use of self-expanding metal stents to treat malignant gastric-outlet, duodenal, and small intestinal obstructions. PMID- 10388861 TI - Metallic stenting for colorectal obstruction. AB - Intestinal obstruction is a major complication of colorectal cancer. Acute surgical decompression frequently requires subsequent operative interventions and is associated with mortality in more than 3% of the cases. Transanal metallic stenting is now possible to perform on an outpatient basis, thus providing quick symptom relief and the opportunity to cleanse the bowel for work-up or surgery. This stenting provides temporary relief for patients and is gaining acceptance rapidly. Randomized controlled trials are needed to help pinpoint refinements that can minimize the difficulty of stent insertion. PMID- 10388862 TI - Endoscopically placed Gianturco endoprosthesis in the treatment of malignant and benign biliary obstruction. AB - Since the introduction of the Z-stent by Cesare Gianturco in 1985, the prominent role of endoscopically placed transpapillary endoprostheses as the treatment of choice to relieve malignant biliary obstruction has stimulated much interest and research in the evolution of his initial design. This article reviews the efficacy and limitation of prior Z-stent models in their attempt to relieve malignant and benign biliary obstructions and previews improvements in the design currently being evaluated in a large, multicenter trial. PMID- 10388863 TI - The wallstent for malignant biliary obstruction. AB - The self-expandable metal Wallstent was introduced in late 1980 for the treatment of malignant biliary strictures. The technique of insertion in distal and hilar bile duct strictures is described in detail. The author presents the reported data and discusses the indications for the stent's use. This stent still does not fulfill all the criteria for an ideal stent; further improvements should be made. PMID- 10388864 TI - The endocoil stent for malignant biliary obstruction. AB - The Endocoil (Instent, Inc., Eden Prairie, MN), first introduced in 1993, is a self-expandable nitinol stent made of a coil spring of nickel-titanium alloy. Advantages of the Endocoil in patients with malignant biliary obstruction were thought to include increased radial force with more rapid stricture dilation, inhibition of tumor ingrowth caused by the stent's coil framework with closed approximation of loops, and the possibility for endoscopic removal. Unfortunately, in subsequent reports of patients undergoing Endocoil placement, there have been significant problems with incomplete expansion or twisting during deployment, stent migration, and tumor ingrowth. This article reviews the available literature regarding Endocoil placement for malignant biliary obstruction and addresses the authors' experience at a tertiary referral center. PMID- 10388865 TI - The wallstent for benign biliary obstruction. AB - Endoscopic stent placement for biliary strictures is well-established. The majority of biliary strictures, particularly benign, are treated with plastic stents. The role of permanent expandable stents in the treatment of benign biliary strictures is still questionable. When expandable stents are used, it appears that those with a wider mesh have a better long-term response. In this article, the authors present data on the Wallstent for benign biliary strictures. PMID- 10388866 TI - The endocoil for benign biliary obstruction. AB - This article evaluates the use of the Endocoil in patients with benign biliary obstruction. It contains detailed information regarding the practicalities of insertion and retrieval of the Endocoil stent and examines the evidence for its use in this group of patients. The author discusses complications and how to avoid them, and provides recommendations for the future use of the Endocoil. PMID- 10388867 TI - Effect of intravenous calcitonin gene-related peptide antagonist on the laryngeal chemoreflex in piglets. AB - The laryngeal chemoreflex (LCR) is characterized by mixed apnea and cardiovascular instability and is elicited by applying water to the laryngeal mucosa of developing animals. The LCR may be fatal in very young animals, and the reflex has been postulated as a possible mechanism of sudden infant death syndrome. Several antagonists have been found to alter the severity of the LCR, but the primary neurotransmitters involved in mediating the reflex response are not yet well understood. This study investigates the effect, on the LCR, of the pharmacologic antagonism of calcitonin gene-related peptide (alphaCGRP), a neurochemical found in abundance in the mammalian laryngeal mucosa and its innervating system. The LCR was elicited in 10 mixed breed piglets, 17.7 days of age (12 to 22 days), before and during infusion of alphaCGRP 8-37, a pharmacologic inhibitor of alphaCGRP, and cardiorespiratory and laryngeal responses were compared. The duration of obstructive apnea decreased from 17.9 to 9. 8 seconds (P < 0.03) in the presence of alphaCGRP 8-37. Mean central apnea did not change for the group (P > 0.05), although it was completely inhibited in 2 animals. Cardiovascular changes were not significantly altered by the alphaCGRP inhibitor. alphaCGRP appears to play a regulatory role in the apneic response of the LCR, particularly its obstructive component, but not the cardiovascular response. PMID- 10388868 TI - Systemic reactivation of otitis media with effusion in a rat model. AB - OBJECTIVE: This study addresses the interaction of bacterial antigens, specifically peptidoglycan-polysaccharide (PG-PS) and lipopolysaccharide (LPS), in the induction and reactivation of mucoid middle ear effusions. METHODS: Twenty seven rats underwent eustachian tube obstruction before inoculation of the middle ear bulla with PG-PS. Three weeks later, after resolution of all middle ear effusions, 6 rats were randomly selected and euthanized as the first control group (control I). The remaining 21 animals were randomly assigned to 3 groups that received intravenous injections of Krebs Ringer (control II), PG-PS, and LPS, respectively. These rats were euthanized 2 days after intravenous injection. Middle ear mucin production and histologic changes were measured in all animals. RESULTS: The mean concentrations of mucin were 0.94 +/- 0.52 mg/mL, 0.41 +/- 0.87 mg/mL, 16.33 +/- 3.67 mg/mL, and 1.15 +/- 0.41 mg/mL in the control I, control II, PG-PS, and LPS groups, respectively. Thus the mean concentration of mucin in the middle ear lavage samples was significantly greater in rats that were injected intravenously with PG-PS than in rats in other groups (P < 0.05). Histologic analyses demonstrated a greater degree of goblet cell hyperplasia in the PG-PS group than in other groups. CONCLUSIONS: This is the first animal model of recurring otitis media with effusion in which a systemic injection of PG-PS was used to reactivate a middle ear effusion in rats previously primed with a transtympanic injection of PG-PS. This study suggests that after otitis media with effusion has resolved, it may be reactivated by the presence of bacterial antigens and/or cytokines in the systemic circulation. PMID- 10388869 TI - Approach to the vestibular patient and driving: A patient perspective. AB - OBJECTIVES: To learn about the impact of dizziness on driving from a patient perspective and to present an approach to the vestibular patient and driving. DESIGN: An anonymous questionnaire completed by 265 dizzy patients at 3 different centers. RESULTS: The participants were experienced drivers who needed to drive to function normally (83%). Those with constant or severe dizziness comprised a higher risk group of drivers. Although few had ever been warned not to drive, 52% said that if they were warned to stop driving, they would not. Most thought that it was the doctor's role to report unsafe drivers to the authorities (P < 0.001, chi2 = 87.2670). CONCLUSIONS: The diagnosis of a vestibular disorder should not alone be grounds to suspend a patient's driver's license. Legislation should be amended to better reflect the views of doctors and patients alike. PMID- 10388870 TI - Relevant issues in revision canal-wall-down mastoidectomy. AB - The role of canal-wall-down (CWD) versus intact-canal-wall (ICW) mastoidectomy in the treatment of chronic otitis media with or without cholesteatoma remains a debated issue. Although past conventional wisdom has held that CWD mastoid surgery leads to a "safe" ear and is technically less demanding than the more controversial ICW mastoidectomy, this is often not the case. Although our preference is an ICW technique when possible, 47 of 109 (43%) mastoid procedures for chronic otitis media performed between January 1993 and June 1996 involved a CWD mastoid cavity. More than two thirds of these procedures (32 of 47) represented revision surgery, the most common indication being a poorly contoured, preexisting CWD mastoidectomy with persistent otorrhea. A dry, well epithelialized ear was obtained in 90% of cases. Our preferred method of ossicular reconstruction (double cartilage block ossiculoplasty) is detailed, and hearing results according to American Academy of Otolaryngology Head and Neck Surgery guidelines are reported. PMID- 10388871 TI - Effects of sound pressure levels on fundamental frequency in tracheoesophageal speakers. AB - The quality of tracheoesophageal (TE) voice after laryngectomy is dependent on numerous factors. The relative contribution of specific variables is not well described. To evaluate the modulation of fundamental frequency (F0) pitch in TE speakers after total laryngectomy and voice restoration, we instructed 11 TE speakers to complete a series of vocal tasks under standardized conditions. All patients underwent standard laryngectomy with appropriate neck dissection and full-course radiation therapy. Each speaker produced 4 sustained phonations at distinct levels. Sound pressure level (SPL) intensity and F0 measurements were taken for each level. Statistical analysis to assess the relative effect of SPL on F0 demonstrated a statistically significant effect. However, the knowledge of SPL allowed only a 9% greater prediction of F0, indicating that factors other than SPL are integral to the modulation of F0 by TE speakers. These data lend further support to the theory that TE voice is an active process incorporating the myoelastic properties of the pharyngoesophageal segment as well as the properties of aerodynamic flow. PMID- 10388872 TI - Penicillin-nonsusceptible pneumococcus in acute otitis media in New York City. AB - OBJECTIVE: To determine the proportion of children with acute otitis media (AOM) presenting in our catchment area in New York City who were infected with nonsusceptible Streptococcus pneumoniae and to determine the susceptibility of these organisms to penicillins and other antibiotics commonly used to treat AOM. SETTING: Ambulatory clinics and the emergency department of a tertiary care, inner-city medical center. PATIENTS: During a 2-year period from 1993 to 1995, 115 children (aged 6 months to 12 years) with AOM underwent tympanocentesis. Patients did not receive antibiotics for at least 1 week before tympanocentesis. RESULTS: Thirty-one children were infected with S pneumoniae, and 83.9% of isolates were susceptible to penicillin. Of the 16.1% strains that were nonsusceptible, most (4 of 5 strains) were intermediately resistant, and only 1 exhibited high-level resistance to penicillin. Of all the cephalosporins tested, only cefotaxime had consistent activity against the intermediately resistant strains. Notably, all nonsusceptible pneumococci were inhibited by macrolides. CONCLUSIONS: This study provides unique reference data for nonsusceptible Streptococcus pneumoniae in children with AOM and documents that newer cephalosporin agents are not active against all of these strains. PMID- 10388873 TI - Speech perception results in children with cochlear implants: contributing factors. AB - Speech perception test results were obtained from a group of 40 pediatric cochlear implant users. Half of the children participated in oral-only habilitation programs, which included both traditional oral and auditory-verbal approaches, and half participated in programs that used a combination of oral and manual communication referred to as total communication (TC). Analysis of the scores showed that children enrolled in oral-only habilitation programs scored significantly higher on the speech perception measures than did children who were enrolled in total communication based programs. These results were inconsistent with those of other reports, which suggested that communication methods had little effect on implant outcomes. To further examine the reasons for the differences in performance, we analyzed 7 additional factors, including length of implant use, age at surgery, device type, socioeconomic status, bilingualism, school setting, and participation in private therapy, which may affect implant performance. Multiple-regression analysis again showed communication mode to be the factor most highly correlated with speech perception abilities among this group of children. PMID- 10388874 TI - Permeability of tympanotomy tubes to ototopical preparations. AB - Myringotomy with tube placement is the most frequently performed otolaryngologic procedure in the United States, and purulent otorrhea after this procedure is not uncommon. Believing that ototopical preparations have proved useful in treating this malady, our group was compelled to examine whether and in what quantities these preparations are able to penetrate tympanotomy tubes. To this end, models were constructed and in vitro testing carried out with several popular formulations. PMID- 10388875 TI - Efficacy of a eutectic mixture of local anesthetics as a topical anesthetic in minor otologic procedures. AB - A eutectic mixture of local anesthetics (EMLA), prepared as a cream, is an oil-in water emulsion of 2 anesthetic agents lidocaine and prilocaine. Several clinical applications of EMLA cream, its effectiveness as a topical anesthetic, and its safety profile have been previously reported. We report our experience with EMLA cream in 17 adult and 24 pediatric patients. We find EMLA to be the preferred anesthetic for performing minor outpatient otologic procedures in adults. We also find EMLA to be a safe, well-tolerated alternative to general anesthesia in some pediatric patients. Potential cost savings of EMLA cream during pediatric myringotomies in the clinic are also discussed. PMID- 10388876 TI - Healing large tympanic membrane perforations using hyaluronic acid, basic fibroblast growth factor, and epidermal growth factor. AB - Large tympanic membrane perforations usually require a surgical tympanoplasty for closure. Reducing surgical costs and risks has encouraged investigators to examine nonsurgical office procedures for healing these perforations. Growth accelerators are the most promising agents. We study here the closure of large acute perforations using weekly applications of 1 mg of 1% hyaluronic acid (HA), 0.4 microg basic fibroblast growth factor (bFGF), or 1.0 microg epidermal growth factor (EGF) directly to the tympanic membranes of the experimental ears. Control ears were treated with 0. 1 mL Vasocidin. Complete closure was obtained in 100% of the ears treated with HA and EGF and 85.7% of those treated with bFGF by day 21, compared with 63.6% of the controls by day 32. Moderate-to-severe ipsilateral and contralateral external canal hypertrophy was noted in 14.2% and 37.5% of the ears treated with bFGF and HA, respectively, but was not seen in ears treated with EGF or in the control group. PMID- 10388877 TI - Tinnitus severity, loudness, and depression. AB - Answers to questionnaires filled out by 436 patients who visited our tinnitus clinic were analyzed. Patients were asked to report the presence or absence of depression and to rate the loudness and severity of their tinnitus. Responses to questions about tinnitus loudness and severity from 121 patients who reported current depression were compared with responses from 285 patients who reported no history of depression. There was no significant difference in reported loudness of tinnitus between patients with and without depression. However, patients with current depression scored significantly higher than patients without depression on all 12 questions relating to tinnitus severity. We conclude that depression and tinnitus severity are linked in some patients. Treatment of depression with medications and psychotherapy is likely to reduce tinnitus severity for many of these patients. PMID- 10388878 TI - Comparison of optokinetic nystagmus elicited by full versus partial visual field stimulation: diagnostic implications. AB - Optokinetic nystagmus (OKN) testing is one method to determine central vestibular dysfunction. OKN may be elicited by partial visual field stimulation with a light bar (OKN-ENG) or by full visual field stimulation with rotating stripes in a rotational chair test booth (OKN-RVT). OKN-ENG and OKN-RVT were elicited in 36 healthy subjects and 48 patients with known peripheral or central vestibular disorders. Abnormal test results suggested central pathology in 29 of 36 healthy subjects with OKN-ENG versus 1 of 36 with OKN-RVT. Twenty-eight of 33 patients with peripheral pathology demonstrated abnormal OKN-ENG findings, whereas 4 of 33 had abnormal OKN-RVT results. Thirteen of 15 patients with central vestibular disorders had abnormal OKN-ENG, whereas 7 of 15 had abnormal OKN-RVT. Sensitivity and specificity of OKN-ENG were 86.7% and 17.4% versus 46.7% and 92.7%, respectively, for OKN-RVT. These findings were statistically significant (P < 0.00001). OKN elicited by full visual field stimulation (OKN-RVT) is a more accurate indicator of central disease than OKN elicited by partial visual field stimulation (OKN-ENG). The use of OKN-ENG to identify central vestibular dysfunction is questionable. PMID- 10388879 TI - Patterns of nodal metastasis and surgical management of the neck in supraglottic laryngeal carcinoma. AB - BACKGROUND: Appropriate management of the clinically negative (N0) neck in supraglottic laryngeal cancer continues to be an area of controversy in head and neck surgery. Our treatment policy has been aggressive surgical management even in the clinically N0 neck. METHODS: Between 1971 and 1991, 104 patients had the primary diagnosis of supraglottic laryngeal cancer. Ninety of these patients received their treatment at Roswell Park Cancer Institute and are the subject of this retrospective review. RESULTS: All neoplasms included in this study were squamous cell cancers. The most common subsite involved with tumor in our series was the epiglottis, followed by the aryepiglottic folds and false cords. Supraglottic laryngectomy was performed of 29% of the cases; the remainder received total laryngectomy. Thirty-six percent of the patients had pathologic stage I/II disease, and 64% had stage III/IV. The 5-year survival rates were 100%, 81%, 73%, and 63% for stages I through IV, respectively. Fifty-seven patients had clinically N0 disease at presentation; of these 34 underwent elective neck dissection, and the remaining 23 patients were observed. Of those patients receiving neck dissection, 30% (n = 10) were found to have histologically positive disease, and of the 23 patients observed, 30% (n = 7) had histologically positive regional (neck) disease. Of the 17 clinically N0 and pathologically N+ patients, 82% (14 of 17) had involvement of level I (submandibular triangle), and 100% had involvement of level II. The incidence of bilateral disease in the clinically N0 patient was 44%. There were no local failures. CONCLUSIONS: There is a high incidence of occult regional disease even in early-stage supraglottic squamous cell carcinoma of the larynx. In the surgical management the clinically N0 neck, we presently recommend bilateral neck dissection of levels I through IV to adequately address those regions at highest risk for occult disease. PMID- 10388881 TI - Long-term impact of functional endoscopic sinus surgery on asthma. AB - Using objective and subjective criteria, we performed a study to assess the long term impact of functional endoscopic sinus surgery (FESS) in patients with chronic rhinosinusitis and asthma at an average follow-up of 6.5 years. One hundred twenty patients who underwent FESS for chronic rhinosinusitis were followed up for an average of 6.5 years (range 6.0 to 10.6 years). Seventy-two (60%) patients responded to a follow-up questionnaire, and 30 (42%) of them reported a history of asthma. Subjective levels of improvement and assessments of medication need were evaluated and statistically assessed with parametric and nonparametric methods. Of these 30 patients, 27 (90%) reported that their asthma was better than it had been before FESS, 6.5 years ago. Average reported improvement increased from 49% at 1.1 years after surgery to 65% at 6.5 years after surgery. Asthma attacks declined in 20 of 27 (74.1%). Medication use for asthma showed similar improvement, with approximately half reporting less inhaler usage and nearly two thirds reporting less oral steroid use. This study demonstrates that a combination of FESS, careful postoperative care, and appropriate medical therapy for chronic rhinosinusitis has a favorable long-term effect on asthma in patients with symptomatic chronic sinusitis. In this study asthma severity, frequency of attacks, and medication need were all improved. PMID- 10388880 TI - Auricular carcinoma with temporal bone invasion: outcome analysis. AB - Invasion of the temporal bone by cutaneous carcinoma of the auricle and periauricular skin is an ominous prognostic sign. Management includes aggressive resection of cutaneous disease as well as resection of temporal bone to obtain a medial margin. Analysis of data from 21 patients with temporal bone invasion caused by cutaneous malignancy is presented. Overall survival is approximately 63%. Cumulative survival is significantly decreased in patients with squamous cell carcinoma when compared with other invasive malignancies. Univariate and covariate analyses demonstrate that nodal status, positive microscopic soft tissue margins, and persistent perineural disease at the skull base did not significantly affect survival in this series. There is a trend toward increased survival in patients receiving postoperative radiation in this series. PMID- 10388882 TI - Incidence of abnormal laryngeal findings in asymptomatic singing students. AB - OBJECTIVE: Abnormalities in the mucosal lining of the vocal folds may interfere with the normal vibratory patterns and result in vocal limitations, especially for singers whose demands are great. A prospective, longitudinal study was undertaken to investigate the incidence of laryngeal abnormalities in asymptomatic singing students. METHODS: Sixty-five singing students at the school of music underwent videostroboscopic evaluation and completed a comprehensive questionnaire. Videos were rated by 3 experienced clinicians, and interrater reliability was calculated. Results were correlated with demographic factors, background medical history, and singing history. RESULTS: Five students (8.3%) exhibited early signs of benign vocal fold lesions (2 with nodules and 3 with cysts). A high incidence of posterior erythema (n = 44; 73.4%), suggesting possible reflux, was found. CONCLUSIONS: A surprisingly high number of otherwise asymptomatic singing students demonstrated abnormal laryngeal findings. Their relationship with vocal performance will be addressed as well as implications for preventative measures. PMID- 10388883 TI - Calvarial bone graft harvest in children. AB - Bone grafts are occasionally required in the reconstruction of bony defects in the pediatric population. Strong recommendations have existed in the past toward the use of principally inner table bone grafts in children. In this retrospective series with an upper age limit of 14 years, outer table calvarial bone grafts were used as the material of choice for bony reconstructions. There were no complications relative to the outer table graft harvest in any of these 12 patients. Discussion of harvest techniques in different pediatric age groups will be reviewed. PMID- 10388884 TI - Acute effects of palatopharyngoplasty on airway collapsibility. AB - OBJECTIVE: The lateral pharyngeal walls contribute to obstruction in obstructive sleep apnea. These structures may be unaffected by uvulopalatopharyngoplasty. This was evaluated by retrospective review of upper airway observations after palatopharyngoplasty. METHODS AND PATIENTS: The retropalatal airway was endoscopically observed intraoperatively after each procedure in 7 patients. The airway was dilated with nasal continuous positive airway pressure. RESULTS: Transpalatal advancement pharyngoplasty increased the area 120% (P = 0.001), and closing pressure decreased 9.2 cm H2 O (P < 0. 01). The maximal anteroposterior length (MAX-AP) and maximal lateral radius increased 90% (P = 0.01) and 60% (P < 0.001), respectively. MAX-AP changed in 2, both increased in 4, and maximal lateral radius increased in 1 patient. The closing pressure change correlated with airway size (r 2 = 0.44, P < 0.05); airway shape was associated with change in MAX-AP (r 2 = 0.51, P < 0.07). CONCLUSIONS: Both the anteroposterior and lateral wall dimensions are altered by palatopharyngoplasty techniques, which increase retropalatal airway size. This is not limited to facial advancement surgery. PMID- 10388885 TI - Voice target time in Parkinson's disease: A preliminary report. AB - A target-matching paradigm was developed to assess the vocal equivalents of reaction and movement time in Parkinson's disease. Six patients with Parkinson's disease and 6 age- and gender-matched control subjects were asked to enunciate /pa/ to reach a target frequency and intensity level in response to a light stimulus. The stimulus and acoustic responses were simultaneously recorded. Measures included laryngeal reaction time, time between stimulus and phonation onset; frequency voice target time, time from phonation onset to target level of frequency; and amplitude voice target time, time from phonation onset to target level of intensity. The 2 subject groups were significantly differentiated by laryngeal reaction time (t = 299.67, df = 10, P = 0.005) and frequency voice target time (t = 148, df = 10, P = 0.014). These data suggest voice target time is a viable tool for assessing the effects of neurologic disorders on voice execution in Parkinson's disease. PMID- 10388886 TI - Effect of the Palmaz balloon-expandable metallic stent in the trachea of pigs. AB - Endoscopically placed airway stents offer a viable option in primary or adjunctive treatment of severe pediatric tracheobronchial stenoses. Optimistic clinical reports substantiate the need for experimental studies to more effectively evaluate their clinical role. Development of an animal model comparable with the pediatric airway, amenable to endoscopic instrumentation, and capable of assessing effect on growth was the purpose of this pilot project. Nine 4-week-old piglets underwent endoscopic midtracheal placement of the balloon expandable Palmaz metallic stent. Initial expansion and stent position were verified fluoroscopically and by direct videobronchoscopy. Serial endoscopic examination and stent reexpansion were performed 2 and 4 weeks after stent insertion. Animal weight, clinical tolerance, tracheal growth, and stent integrity were observed. Tracheal inflammation was evaluated grossly and by objective histopathologic criteria. Successful endotracheal stent placement and expansion were accomplished in all piglets. One pig died of anesthesia complications less than 24 hours after stent insertion. The remaining pigs exhibited excellent clinical tolerance through experiment completion. No detrimental effect on growth was noted, and effective dilatation of the stented tracheal region was observed. Stent incorporation was evident with significant mucosal ingrowth. Inflammation in the form of nonobstructing granulation tissue was present, and no evidence of necrosis or cartilage invasion was evident. The piglet trachea appears to be an excellent model for evaluation of expandable metallic airway stents in management of congenital and acquired tracheobronchial stenoses. PMID- 10388887 TI - Higher rates of pressure decrease in inflamed compared with noninflamed middle ears. AB - Recent clinical trials have renewed interest in middle ear inflation as a treatment for otitis media with effusion. However, air inflation in human beings with significant negative middle ear pressures was shown to be followed by a rapid pressure decrease to approach the preinflation values. In this experiment, the middle ears of anesthetized rhesus monkeys with unilateral inflammation were inflated at different times with air or N2, and pressures were recorded by tympanometry until they had stabilized or the animal had recovered from anesthesia. The results for air inflations reproduced those reported for human beings with negative pressures. Similarly, after N2 inflation a significantly greater rate of pressure decrease and significantly lesser terminal pressures were observed for inflamed ears when compared with the contralateral control ears. However, the rate of pressure decrease and the magnitude of the pressure drop were dampened by sequential N2 inflations. These observations have clinical implications with respect to the efficacy of inflation as a treatment for otitis media with effusion. PMID- 10388888 TI - Management paradigms for posterior epistaxis: A comparison of costs and complications. AB - OBJECTIVES: Posterior epistaxis is a common otolaryngologic emergency. Management is controversial because of the many treatment options available. These options vary in efficacy, rates of complications, and cost. Posterior nasal packing is the medical management most frequently used to control posterior epistaxis. It is associated with major complications, including stroke, myocardial infarction, arrhythmias, and death. Because of these potential complications, many otolaryngologists monitor patients with posterior nasal packing in the intensive care unit (ICU). However, the level of care used to monitor these patients is variable, and standards have not been established. METHODS: From 1991 to 1997, 46 patients had posterior nasal packing placed to control epistaxis. Management, complications, and hospital charges were analyzed. RESULTS: Six patients (13%) were admitted to the ICU, 2 (4%) were admitted for telemetry monitoring, and 38 (83%) were sent to the ENT ward for continuous pulse oximetry. Four major complications occurred (1 episode of syncope [emergency department], 2 arrhythmias [ICU], and 1 death [hospice]). Twenty-six patients were treated with posterior packing in the ENT ward, at a mean cost of $2988. Fourteen patients underwent intervention (5 ligations, 6 endoscopic cauterizations, and 3 angiograms), with a mean cost of $5482. Six patients spent time in the ICU, with a mean cost of $8242. Patients treated in the ENT ward had significantly lower costs than those undergoing intervention (P = 0.017) or those admitted to the ICU (P = 0.020). CONCLUSION: We propose that most patients with posterior epistaxis can be treated in specialized ENT wards. This can be done without increasing complications and with significantly decreased costs. PMID- 10388889 TI - Posterior wall augmentation for treatment of velopharyngeal insufficiency. AB - Velopharyngeal insufficiency (VPI) can be treated surgically with various operations. This article describes the use of a superiorly based folded pharyngeal flap for posterior wall augmentation to treat VPI. This is a retrospective study indicating that a folded flap to augment the posterior wall is likely to be as effective as other surgical techniques to treat small velopharyngeal gaps. Patients selected for this procedure had very good velar motion. Postoperative nasometric zoo passage scores improved by an average of 18 over preoperative scores. Additionally, a correlation was found between age and nasometry improvement after surgery. Younger patients did better. Patients in whom VPI was caused by adenoidectomy did well. The 2 syndromic patients did not do as well when treated with this type of operation. PMID- 10388890 TI - Critical pathways in anterior cranial base surgery. AB - New advances in anterior cranial base surgery have dictated the need for a comprehensive, multidisciplinary approach in the treatment of lesions of this area, necessitating multiple modes of diagnostic and surgical techniques. Traditional consideration of the complex problems presented by neoplastic involvement of the anterior cranial base predicated on isolated syndrome analysis is no longer sufficient to adequately assess tumor pathology. To address these complex problems, we discuss a method of localization of pathology based on anatomic structure and function as well as the corresponding surgical approach to the anterior cranial base. PMID- 10388891 TI - Chisel tip suction cautery device for tonsillectomy. PMID- 10388892 TI - Intraosseus zygomatic hemangioma. PMID- 10388893 TI - Y modification of the Fisch meatoplasty. PMID- 10388894 TI - Non-Hodgkin's lymphoma presenting as facial swelling and nasal obstruction in a pediatric patient. PMID- 10388895 TI - Viral cochleitis with gadolinium enhancement of the cochlea on magnetic resonance imaging scan. PMID- 10388896 TI - Long-term study of quality and safety of osseointegration for the retention of auricular prostheses. AB - The aim of this article is to describe the safety and quality of the osseointegrated implant technique for the retention of craniofacial prostheses, to present a protocol for collection of clinical data, and to discuss the impact of the procedure on the patient quality of life. A protocol was designed and used to study patients who had received auricular prostheses consecutively since 1979 at our department. The patients were asked to answer a questionnaire designed to describe symptoms and problems specific for someone wearing an auricular prosthesis. In total, 99 patients received 107 prosthetic ears (8 patients had bilateral defects) retained on 309 implants (2 to 4 implants/ear). Patients of all ages were represented, and only 9 discontinuities were reported. Most patients (95%) wear their prosthesis every day, in most cases more than 10 hours/day. The follow-up period ranged from 1 to 12 years, giving a total of 2624 postoperative observations of implants, with a 3% incidence of significant skin reaction. We conclude that the surgical technique for auricular prostheses retained on osseointegrated implants is simple and associated with a low rate of peroperative and long-term complications. It offers a high degree of stability and aesthetic satisfaction. PMID- 10388897 TI - Cerebellar encephalomalacia on magnetic resonance imaging after removal of acoustic tumor. AB - MRI is widely used for postoperative surveillance of patients undergoing surgery for removal of acoustic neuroma. The purpose of this study was to investigate the frequency and pattern of postoperative changes in the cerebellum and brain stem on MRI after removal of acoustic neuroma. A retrospective study was conducted in 30 consecutive patients who underwent postoperative MRI between 1994 and 1995. The timing of the scans after surgery ranged from 12 months to 10 years. T2 weighted turbo spin-echo images revealed cerebellar encephalomalacia in 17 of 30 cases. Cerebellar encephalomalacia was found more consistently in patients who had large tumors and was more frequent after the suboccipital approach. Encephalomalacia is largely caused by gliotic changes in the adjacent cerebellar tissues after tumor removal. PMID- 10388898 TI - Postoperative hypertension effect of carotid sinus denervation. AB - Postoperative arterial blood pressures were monitored in 43 patients who had undergone bilateral neck dissection during a 6-week period at Ankara Numune Hospital's IInd Otorhinolaryngology Department. During the first operations, all cases received carotid sinus denervation, whereas no denervation was done for the opposite side dissections held 6 weeks later. Study and control groups were composed of the same patients to achieve an objective outcome for the risk of postoperative hypertension. Hypertension was observed in 10 (23%) of 43 patients after the first operations and 12 (28%) of 43 patients after the opposite side dissections, for which no carotid denervation was done. The difference between the rates was insignificant statistically. PMID- 10388899 TI - Minimally invasive surgery for posterior glottic stenosis. AB - Posterior glottic stenosis most commonly results from prolonged endotracheal intubation. The tube causes decubitus and perichondritis with a consequent scar tissue formation in the posterior commissure that often limits the abduction of the vocal cords. Many different surgical methods are known for the treatment, but in most cases temporary tracheostomy is required. We recommend a minimally invasive method to avoid tracheostomy, which is a very inconvenient state for the patient. The scar of the posterior commissure is excised endoscopically with the CO2 laser, and a modification of the endoextralaryngeal vocal cord laterofixation described by Lichtenberger is used to lateralize 1 or both vocal cords until the posterior commissure is completely reepithelialized. In this article we report on the first 5 cases. All patients had satisfactory airways immediately after the laterofixation procedure, which proved to be stable later on as well. In the cases of moderate stenosis, further scarring was prevented, and after the healing of the mucosa in the posterior glottic area, the laterofixation sutures were removed. The vocal cord mobility was recovered in the cases in which the cricoarytenoid joint was not fixed. In 1 case of severe stenosis (bilateral cricoarytenoid joint fixation), the procedure yielded only partial improvement. PMID- 10388900 TI - Self-retaining ear speculum: Thudicum design. PMID- 10388902 TI - Internal carotid artery thrombosis and Horner's syndrome as complications of parapharyngeal abscess. PMID- 10388901 TI - Traumatic pseudoaneurysm of the external carotid artery with parotid mass and delayed facial nerve palsy. PMID- 10388903 TI - Primary malignant melanoma of the tongue. PMID- 10388904 TI - Obstructing laryngeal granuloma. PMID- 10388905 TI - Follow-up clinic and ambulatory blood pressure in untreated white-coat hypertensive patients (evaluation after 2-5 years). AB - BACKGROUND: It is still under debate whether subjects with persistently elevated clinic blood pressure but normal ambulatory blood pressure, [white-coat hypertensives (WCH)] have a higher propensity for further development of ambulatory hypertension. METHODS: We prospectively evaluated for 3.5 years (from 26 up to 59 months) the transition of clinic and ambulatory blood pressure values in 36 untreated subjects (17-65 years) with WCH (clinic blood pressure > 140/90 and awake ambulatory blood pressure < 132/84 mmHg and without any other major cardiovascular risk factors) and of 52 clinic and ambulatory normotensive subjects (clinic blood pressure < 140/90 mmHg and awake blood pressure < 132/84 mmHg, 24-61 years). RESULTS: Average values of clinic blood pressure, 24 h blood pressure and awake blood pressure values did not differ from baseline to the end of the follow-up period in both populations. Development of ambulatory hypertension occurred in four out of 36 (11%) subjects with WCH and in three out of 52 (6%) normotensives if defined by awake blood pressure >/= 140/90 mmHg and in eight out of 36 (22%) subjects with WCH and in eight out of 52 (15%) normotensives if defined by awake blood pressure >/= 132/84 mmHg, respectively. Patients who reached ambulatory hypertension had baseline awake blood pressure values within the upper quintile of distribution of blood pressure in their respective group. CONCLUSION: After an average of 3.5 years of follow-up, a transition to ambulatory hypertension occurred in a similar way in normotensives and subjects with WCH without any other cardiovascular risk. A small propensity for the development of sustained hypertension may affect patients with higher initial ambulatory blood pressure values. Although a slow evolution towards sustained hypertension cannot be excluded in subjects with WCH, these findings suggest that this transition might be similar to that in normotensive subjects. PMID- 10388906 TI - Ignaz Semmelweis and the conquest of puerperal sepsis. PMID- 10388907 TI - Errors in transfusion medicine. Scope of the problem. AB - Error is ubiquitous whenever humans are involved in a process. Fortunately, most transfusion-related errors are benign. However, the risk of death due to acute hemolytic transfusion reaction rivals that of human immunodeficiency virus transmission and administration of the wrong blood or of blood to the wrong recipient has occurred at many facilities. Most blood misadministration errors are caused by failure to identify the recipient and blood unit adequately, although phlebotomy errors and blood bank errors also contribute. Many errors are multifactorial and may reflect underlying systems defects. Noncompliant specimen labels may be a cue to an increased risk of phlebotomy error. Autologous blood is not immune from error and poses infectious disease risks as well as the risk of hemolytic transfusion reaction; also, perioperatively recovered blood may pose a risk of air embolism if improperly handled. PMID- 10388908 TI - The quality systems approach. AB - Health care as an industry is going through an evolution similar to that experienced by manufacturing and other service sectors of the world economy. When competition threatens, the traditional management approach is slash-and-burn to manage costs. This approach affects people, services, and facilities. When the slash-and-burn method runs out of fuel, organizations start to merge, thinking that bigger is better. Just getting bigger only looks like change, with a new cast of fewer people and organizational structure. The root cause for existing problems has not been fixed. The only real solution, as manufacturing and services learned, is to understand and improve work processes to ensure that the right work is being done in a high-quality manner and at a competitive price. The era of quality systems is just dawning in health care. The focus must remain on quality patient outcomes at a competitive price by improving supporting systems and processes to accomplish these goals. PMID- 10388909 TI - Process improvement in transfusion medicine. AB - Ongoing efforts to decrease costs in the clinical laboratory make continuous process improvement especially important in difficult economic times. Process improvement can result in decreased workload, cost savings, and increased customer satisfaction but is an abstract concept in and of itself. To illustrate the steps of process improvement, we applied them to our blood component retrieval policy. By identifying the problems with the current system, proposing and implementing solutions, and measuring the effects before and after revamping the process, we have been able to show impressive reductions in the number of component retrievals initiated, the number acted on, wasted components, and customer complaints, all of which translate into cost savings. Once the cycle is completed, it begins anew. There must always be continuous process improvement. PMID- 10388910 TI - The "P's and Q's" of quality systems. AB - Quality improvement in blood collection centers has been a priority of regulatory and accrediting agencies for the past several years. The Food and Drug Administration and the American Association of Blood Banks have developed guidelines for quality assurance activities. Inspection programs have focused on evaluation of processes and how they are controlled to assure the safety and efficacy of blood components. A review of Food and Drug Administration enforcement actions shows that all such actions cite similar deficiencies related to management control, personnel training, error, and record management policies. A quality program that includes management commitment to compliance and continuous improvement, defined personnel training, internal audit, and error management policies provides documented evidence to management and regulatory agencies that operations are in control. PMID- 10388911 TI - The costs and consequences of management fads and politically driven regulatory oversight. AB - CONTEXT: The period from the mid-1980s to the present has seen a virtual revolution in how blood collection, testing, and transfusion are carried out. Most of the changes that have occurred represent changes in organization, underlying paradigms of transfusion safety, and approaches to solutions to perceived problems, rather than technical or scientific advances. These changes were instituted with little or no data to support their safety and efficacy, much less their cost-effectiveness. DESIGN: Using data on changes in fees charged by regional blood centers during this period, an estimate was generated of the costs attending the purely administrative and organizational changes in the field of transfusion medicine. RESULTS: The additional expenditures, excluding inflation, are in the range of $226 million per year. This figure does not include changes in infectious disease testing on donated blood, the substantive scientific advance during this period. CONCLUSIONS: In addition to the questions of safety and efficacy, which are traditionally considered prior to clinical or procedural changes of this magnitude, the question remains whether these expenditures are a reasonable trade-off of cost versus benefit. Similar applications of federal regulatory standards intended for manufacturing (eg, good manufacturing practices) and industrial management theory (eg, total quality management) are now being implemented or proposed for hospitals, health care delivery systems, and individual physician's practices. Thus, the case of transfusion medicine may provide some cautionary lessons for future proposed changes in health care delivery and medical practices. PMID- 10388912 TI - Performance improvement in a hospital transfusion service. AB - Blood bankers have always embraced the concepts of quality exemplified by the American Association of Blood Banks' Standards for Blood Banks and Transfusion Services and its accreditation program. The emergence of a quality management system based on a set of quality system essentials represents a natural evolution of these quality initiatives. When fully implemented, a quality system provides a powerful tool with which to improve the quality of both intralaboratory and, importantly, extralaboratory processes related to transfusion medicine. In addition, a functional quality system enhances a transfusion service's ability to meet current and proposed requirements of the American Association of Blood Banks, the Food and Drug Administration, the Joint Commission on Accreditation of Healthcare Organizations, and the National Committee for Clinical Laboratory Standards. PMID- 10388913 TI - Guiding the decision to transfuse. AB - Guiding the decision to transfuse can improve transfusion practices. Effective processes must first identify problem(s) in transfusion practice and then include the attending physician as an educational target. Process improvements that have been shown to be effective include the following: (1) briefly meeting one-on-one with physicians, (2) teaching at scheduled conferences, (3) making daily clinical rounds of patients who receive transfusion, (4) concurrently reviewing orders for transfusion before issue of the blood product, and (5) installing algorithms and guidelines in the operating room. Transfusion practices improved with these process improvements. PMID- 10388914 TI - Assessing blood administering practices. AB - The risk that a red blood cell unit will be associated with an ABO-incompatible transfusion is currently slightly greater than the aggregate risk of acquiring human immunodeficiency virus, human T-cell lymphotropic virus, hepatitis B virus, or hepatitis C virus by transfusion. Since the most common cause for ABO incompatible transfusion is the failure of transfusionists to properly identify a patient or a blood component before a transfusion, transfusion services are encouraged to evaluate and monitor the processes of dispensing and administering blood. In addition, a proposal of the Health Care Financing Administration of the Department of Health and Human Services would require hospitals to use a data driven quality assessment and performance improvement program that evaluates the dispensing and administering of blood and that ensures that each blood product and each intended recipient is positively identified before transfusion. The Los Angeles County+University of Southern California Medical Center assesses the blood dispensing and administering process as proposed by the Health Care Financing Administration. During the fourth quarter of 1997, 85 blood transfusions were assessed for compliance with the Los Angeles County+University of Southern California Medical Center policies and procedures: 55 transfusion episodes had no variance from institutional protocol and 30 had one or more variances. Of the transfusions with at least one variance, 16 had one or more variances involving the identification of the patient, the component, or the paperwork. The remaining 14 transfusions had one or more variances involving other criteria (nonidentification items). The most frequent variance was the failure to document vital signs during the first 15 minutes after a transfusion was started or after 50 mL of a component had been transfused. No variances in patient or blood component identification were noted in nursing units whose staff routinely performed self-assessment of blood administering practices. Based on these findings, a corrective action plan was implemented. Follow-up assessments (n = 63) were conducted after 3 months (during the second quarter of 1998). The compliance with the pretransfusion identification protocol improved from 81% to 95%. The most common reason for noncompliance continued to be a lack of checking vital signs. This report demonstrates the value of using a data-driven program that assesses blood administering practices. PMID- 10388915 TI - Outcomes in transfusion. AB - Outcomes data in medicine can be limited by subjective methodologic issues such as poor selection of end points and use of nonvalidated systems for quality adjustment. Blood transfusion analyses are further complicated by the fact that transfusion seldom is primary therapy but is usually supportive or adjunctive. Thus, much of the outcome data in transfusion medicine are either unavailable or in one of two areas. The first area is prevention of bad sequelae of various cytopenias or factor deficiencies. The second is decreasing adverse effects of transfusion itself. A different useful area for outcome and root cause approaches in individual institutions is examining preanalytical and postanalytical processes of their own. Examples are sample labeling accuracy, quality and timeliness of blood suppliers, internal delivery processes and times, and product wastage. Use review can be changed to real time from retrospective time. By reducing complaints about service to objective data, realistic change can be made in internal and external processes. PMID- 10388916 TI - Optimizing the cost-effectiveness of quality assurance in transfusion medicine. AB - Although quality assurance efforts have been integrated into many aspects of American health care, their value has been questioned. They can consume large amounts of resources (monetary and/or temporal), calling into question their cost effectiveness. To improve the yield of quality assurance efforts and limit their consumption of administrative resources, they need to be focused on those aspects of the operation where improvement is needed or where errors are particularly problematic and costly. Just as a quality assurance program needs to define the outcome required of the process being monitored, the outcome of the quality assurance process needs to be defined at the outset; the simplest possible system should then be designed to capture the necessary data to direct improvement. Although quality assurance efforts have been documented to yield substantial savings, their real payback is provided through better control of an operation and more complete knowledge of the status of that operation. PMID- 10388917 TI - Using outlier events to monitor test turnaround time. AB - OBJECTIVES: To determine the causes of excessive test turnaround time (TAT) and to identify methods of improvement by studying reasons for those tests reported in excess of 70 minutes from the time the test was ordered (ie, outliers). DESIGN: Self-directed data-gathering of stat outlier TAT events from intensive care units and emergency departments, with descriptive parameters associated with each event and additional descriptive parameters associated with the participant. PARTICIPANTS: Laboratories enrolled in the 1996 College of American Pathologists Q-Probes program. MAIN OUTCOME MEASURES: Components associated with outlier TAT events and outlier TAT rates. RESULTS: Four hundred ninety-six hospital laboratories returned data on 218 551 stat tests, of which 10.6% had TATs in excess of 70 minutes. Ten percent of stat emergency department tests and 14.7% of stat intensive care unit tests were outliers. Major areas in which delays occurred were test ordering, 29.9%; within-laboratory (analytic) phase, 28.2%; collection of the specimen, 27.4%; postanalytic phase, 1.9%; and undetermined, 12.5%. The type of test performed was a significant factor and was independent of location: Chemistry-Multiple Test appeared most frequently ( approximately 40%), followed closely by Hematology-Complete Blood Count (approximately 20%) and Chemistry-Single Test ( approximately 18%). Factors of outlier TAT components for intensive care unit specimens were identified using statistical modeling and included hour of day, type of health care personnel collecting specimen, performing the test in a stat laboratory, and reason for delay. Outlier rates were not associated with any identified factors. The practice parameters of laboratories with outlier rates in the lowest 10th percentile significantly differed from those with rates in the top 10th percentile in test request computerization, report methods, and ordering methods. CONCLUSIONS: We observed that outlier analysis yields new information, such as type of test and reason for delay, concerning test delays when compared with TAT determination alone. Laboratories experiencing stat test TAT problems should use this tool as an adjunct to routine TAT monitoring for identifying unique causes of delay. PMID- 10388918 TI - Necessity of clinical information in surgical pathology. AB - OBJECTIVES: To examine the frequency and nature of problems caused by inadequate clinical data provided on surgical pathology requisition forms. DESIGN: Participants in the 1996 Q-Probes voluntary quality improvement program of the College of American Pathologists were asked to document prospectively all surgical pathology cases with inadequate information. Inadequate clinical information was defined as the pathologist's need for additional clinical information before a diagnosis could be rendered, regardless of the amount of information already present on the requisition slip. Cases that had no clinical information on a requisition slip were not counted if the lack of history did not hinder diagnosis. The study concluded when 3 months had elapsed or 40 surgical pathology cases were documented. The following data were recorded for each case: anatomic site, type of procedure, nature of disease, method of obtaining additional information, importance of obtained information, and the length of delay in the final diagnosis. PARTICIPANTS: Three hundred forty-one laboratories, 322 of which were from the United States. RESULTS: A total of 5594 cases (0.73%) required additional clinical information for diagnosis (10th through 90th percentile range, 3.01% to 0.08%). Institutions with greater average occupied bedsize, a greater number of cases accessioned per year, and a greater number of pathologists had a lower percentage of cases with inadequate clinical data (P <.05). Sixty-eight percent of these cases had no delay in completion of a case, 16.2% had a delay of 1 day or less, and 15.1% of cases were delayed more than 1 day. In 59.4% of cases, the additional clinical information obtained confirmed the initial diagnostic impression. In 25.1%, the information was not relevant to the pathologic diagnosis. In 6.1% there was a substantial change in the diagnosis or a revised report was issued, and in 2.2% no additional information could be obtained. Specific anatomic sites that correlated with a higher rate of changed diagnoses or revised reports in cases with inadequate information included the small bowel, the bronchus/lung, and the ovary. Resection specimens were also significantly associated with a higher rate of changed diagnoses or revised reports when additional information was obtained, as were malignant neoplasms and therapy-induced changes. CONCLUSIONS: This study establishes an aggregate rate of cases with inadequate clinical information for diagnosis (0.73%) and documents the extent of problems caused by inadequate clinical information. The criticality of appropriate clinical information provided to the pathologist is identified for specific anatomic sites and disease processes and is reflected in changed diagnoses or revised reports. PMID- 10388919 TI - Handling sentinel lymph node biopsy specimens.A work in progress. AB - This article reviews the "state of practice" with regard to sentinel lymph node biopsy, a new and evolving technique currently used most commonly for staging of malignant melanoma and adenocarcinoma of the breast. Sentinel lymph node biopsy has the potential to both increase the accuracy of lymph node sampling as a prognostic tool and to decrease the need for unnecessary and morbid extensive lymph node dissection in such patients. The need for close cooperation and planning involving the surgeon and pathologist is stressed, and gross room tissue handling, radiation safety, microscopic examination, and the use of ancillary diagnostic techniques are discussed. PMID- 10388921 TI - Spherulosis of the breast. A spectrum of municous and collagenous lesions. AB - OBJECTIVE: Collagenous spherulosis of the breast is an uncommon localized pattern of basement membrane material deposition that may be mistaken for atypical proliferations or carcinoma. This report describes 9 cases in which the predominant or exclusive appearance of the spherules was basophilic instead of eosinophilic. DESIGN: The files of all cases of collagenous spherulosis diagnosed at the Armed Forces Institute of Pathology were reviewed to ascertain the frequency of diagnosis. RESULTS: Spherulosis with a predominantly basophilic pattern had a histochemical and immunohistochemical profile similar to collagenous spherulosis and was associated with more collagenous-appearing forms in 7 of 9 cases. Review of 81 cases showed that collagenous spherulosis was correctly diagnosed in 15% of referrals and was mistaken for intraductal or invasive carcinoma in 11% of cases. CONCLUSIONS: Mucinous and collagenous patterns appear to be related forms of spherulosis. They are underrecognized by pathologists and maybe mistaken for atypia or malignancy. PMID- 10388920 TI - Comparison of the Vitek GPS-TB card with disk diffusion testing for predicting the susceptibility of enterococci to vancomycin. AB - OBJECTIVE: To compare the ability of the Vitek GPS-TB card with disk diffusion testing for determining the susceptibility of enterococci to vancomycin. DESIGN: Vitek susceptibility testing was performed using the GPS-TB card and software version R05.03. Disk diffusion susceptibility testing was performed according to National Committee for Clinical Laboratory Standards guidelines. When discrepancies occurred between the interpretation of Vitek and disk diffusion, both tests were repeated and the epsilometer test (E test) and agar screen containing 6 microgram/mL vancomycin were performed. RESULTS: Of 415 isolates tested, 313 were susceptible to vancomycin and 97 were resistant to vancomycin by both test methods. Two isolates were intermediate by Vitek and resistant by disk diffusion, 2 were intermediate by Vitek and susceptible by disk diffusion, and 1 was susceptible by Vitek and intermediate by disk diffusion. All but 1 of these latter 5 isolates (intermediate by Vitek and susceptible by disk diffusion) were available for retesting. On repeat testing, the 2 isolates that were intermediate by Vitek and resistant by disk diffusion were resistant by both methods, the 1 isolate that was intermediate by Vitek and susceptible by disk diffusion was susceptible by both methods, and the isolate that was susceptible by Vitek and intermediate by disk diffusion was also susceptible by both methods. These results were confirmed by E test and agar screen. CONCLUSION: We found the results of the GPS-TB card compared well with disk diffusion. However, isolates with intermediate results by Vitek should be retested using another method, such as the E test. PMID- 10388922 TI - Myelolipoma of the renal sinus. An unusual site for a rare extra- adrenal lesion. AB - Extra-adrenal myelolipomas are rare; approximately 36 cases have been reported to date. We document a case of myelolipoma presenting as a localized mass in the renal sinus of a 66-year-old man. The chief clinical and radiologic differential diagnostic considerations in this case included a malignant renal tumor arising in the hilum. The patient was being investigated for recurrent urinary tract infections and vague abdominal pains. Histologically, the lesion showed features characteristic of a myelolipoma. There was also marked chronic inflammation in and around the mass. The uneventful follow-up of 62 months is in keeping with the benign nature of this lesion. This report expands the possibilities of the differential diagnoses of renal hilar neoplasms, particularly in view of the increased use of imaging techniques that are bound to detect many incidental lesions in this region. PMID- 10388923 TI - Oncocytic mucoepidermoid carcinoma of the trachea. AB - We report a rare case of an oncocytic mucoepidermoid carcinoma of the trachea, which presented in a 78-year-old woman with hemoptysis. Oncocytic cells comprised the majority of this low-grade lesion and demonstrated granular cytoplasmic phosphotungstic acid-hematoxylin staining as well as strong immunohistochemical reactivity to antimitochondrial antibody. Most tracheobronchial tumors with oncocytic change are carcinoid tumors. To our knowledge, this is the first oncocytic mucoepidermoid carcinoma of the trachea reported. This diagnosis was facilitated by histochemical and immunohistochemical studies. PMID- 10388924 TI - Collecting duct meningeal carcinomatosis. AB - Collecting duct carcinoma (CDC) is an aggressive primary renal neoplasm that represents a distinct subtype of renal cell carcinoma. Histochemical (eg, mucicarmine) and immunohistochemical (eg, Ulex europaeus) studies, taken in concert with the gross and histologic findings, allow differentiation of CDC from the conventional varieties of renal cell carcinoma in most cases. Collecting duct carcinoma generally pursues a more aggressive course than conventional renal cell carcinoma. Metastases to regional lymph nodes, bone, adrenal glands, lung, and skin have been reported in CDC. We describe the case of a 26-year-old man who presented with a clinical and radiologic impression of multifocal meningioma. Biopsies of the meninges and extracranial soft tissues revealed metastatic adenocarcinoma; subsequent studies suggested metastatic CDC. Ultrasound-guided biopsy was performed on a subsequently identified renal mass, which showed features consistent with CDC. To our knowledge, this is the first reported case of meningeal carcinomatosis due to CDC. The diagnostic features of this tumor are discussed. PMID- 10388925 TI - Blood transfusion-acquired hemoglobin C. AB - Unexpected and confusing laboratory test results can occur if a blood sample is inadvertently collected following a blood transfusion. A potential for transfusion-acquired hemoglobinopathy exists because heterozygous individuals show no significant abnormalities during the blood donor screening process. Such spurious results are infrequently reported in the medical literature. We report a case of hemoglobin C passively transferred during a red blood cell transfusion. The proper interpretation in our case was assisted by calculations comparing expected hemoglobin C concentration with the measured value. A review of the literature on transfusion-related preanalytic errors is provided. PMID- 10388926 TI - Multiple neurofibromas in severe neurofibromatosis type 1. PMID- 10388927 TI - From Pittsburgh to Cleveland: NHBD controversies and bioethics. PMID- 10388928 TI - Moral agency and the family: the case of living related organ transplantation. PMID- 10388929 TI - Cutting bodies to harvest organs. PMID- 10388930 TI - Expectations and outcomes in organ transplantation. AB - Research is needed on the frequency of bad outcomes in transplantation. Allocation policies and professional or institutional self-interest may affect the incidence of bad outcomes, and the need for reform is stressed. Transplant recipients who have had a bad outcome often continue to receive aggressive care. The humanistic care of patients having bad outcomes requires attention to advance directives, discussion with patient and family of alternatives to aggressive treatment, and provision of an option for home hospice care. Finally, it must be reemphasized that the average typical good outcome is extraordinarily good, restoring function of a vital organ, extending and improving quality of life, and sometimes restoring near-normal health. In no way should the fact of bad outcomes reduce our commitment to producing good outcomes. PMID- 10388931 TI - A critique of UNOS liver allocation policy. United Network for Organ Sharing. PMID- 10388932 TI - The ethics of competition in liver transplantation. PMID- 10388933 TI - Recasting the debate on multiple listing for transplantation through consideration of both principles and practice. PMID- 10388935 TI - CQ sources/bibliography. Organ transplantation: shaping policy and keeping public trust. PMID- 10388934 TI - Two steps to three choices: a new approach to mandated choice. PMID- 10388936 TI - Retransplantation and the "noncompliant" patient. PMID- 10388937 TI - The development of bioethics in Mexico. PMID- 10388938 TI - Genetic knowledge and third-party interests. PMID- 10388939 TI - Informed consent in the genetic age. PMID- 10388940 TI - Introduction to the transformation of social experience in Chinese society: anthropological, psychiatric and social medicine perspectives. PMID- 10388941 TI - The transformation of everyday social experience: what a mental and social health perspective reveals about Chinese communities under global and local change. AB - Chinese communities in East Asia are undergoing great economic and social change. The result includes material prosperity but also worsening mental and social health indices. Epidemiological studies clarify the magnitude of these problems and the particular vulnerability of women and the elderly. Ethnographic studies indicate that moral experience is also changing, and that with it subjectivity is being altered in ways that are of concern. And yet it is not clear, in historical perspective, how to assess these changes when they are compared to the extraordinarily difficult experiences of the Chinese over most of the Century. PMID- 10388942 TI - Suicide and social change in China. AB - Using recently available data from China's Disease Surveillance Points system, we estimate that there are over 300,000 suicides in China per year; this makes suicide one of the most important causes of death in the country and makes the suicide rate in China one of the highest in the world. Moreover, the pattern of suicides in China is quite different than in other parts of the world--there are more completed suicides among females than males and rural rates are three-fold urban rates. The lack of reliable suicide data prior to 1987 makes it difficult to determine whether the rates are currently rising, falling, or staying constant. However, reports of suicides in the Chinese press and case studies conducted by the authors suggest (but do not prove) that the high rates of suicide currently experienced are related to the social changes that have occurred with the economic reforms (which started in 1978). Another possible explanation for the high rates of suicide is the large numbers of persons with depressive illness in China who remain untreated. Single-cause models of suicide (i.e., social factors or mental illness) do not do justice to the complexity of the processes involved and, therefore, do not provide useful information about the etiology and prevention of suicide in China or elsewhere. We describe our own dynamic model of suicide that includes five interacting factors which, we believe, collectively determine the suicide rates in a community. PMID- 10388943 TI - Fat, fatigue and the feminine: the changing cultural experience of women in Hong Kong. AB - This paper seeks to demonstrate that rapid economic development in Hong Kong has transformed not only social structures but also Chinese women's subjectivity and bodily experience, thereby producing new forms of identity, aesthetics and aspirations, in addition to novel patterns of distress. Evidence is assembled to show that women's being-thin-yet-feeling-fat and being-active-yet-feeling-tired reflect not so much psychopathology as transformation in embodied moral experience. Because such normative experiences are grounded in the conflicting demands of production and reproduction that recent social transformations have brought to bear on women's lives, "fat" and "fatigue" can be said to embody what it is to become a woman in contemporary Hong Kong. PMID- 10388944 TI - Rural youth and youth culture in north China. AB - This article describes how economic reforms have transformed the local world of village life in North China, specifically the emergence of a rural "youth culture" that reflects urban values and styles. This culture has both challenged and enriched rural China's tradition and its more recent communal structure, a change that reshapes the relations amongst family members of different age cohorts as well as between peasants and cadres, it is argued that this transformation in the Chinese countryside holds opportunities to improve living conditions and unmake the constraints of tradition and the modern nation-state on life chances and everyday experience. The sheer number of rural youth in China suggests that the future impact of this transformation on social realities could be enormous. PMID- 10388945 TI - The changing moral economy of ancestor worship in a Chinese emigrant district. AB - This paper describes the reciprocal influences between Anxi County Fujianese, whose families and clans have migrated to Singapore, and their ancestral villages in Fujian, China. The Singaporeans bring wealth and cultural capital to their poor relations in rural China. Their participation is crucial for local socioeconomic development. Besides bringing material support and globalizing values and lifestyles, they also reinvigorate and transform the local religious tradition. They, in turn, reaffirm and even remake their own ethnic and regional identity. The complex outcome illustrates the fact that China's social change under economic reforms and global influence is, in its huge rural core, not merely a matter of infrastructural, market, and social welfare improvements, but involves exchange and transformation in meanings of rituals and experiences. We can see that kinship and religion are not unchanging aspects of the cultural tradition that are separate from programs of modernity. Indeed, modernity and tradition appear to be inseparable, and they may reveal that the recipe for effective community projects requires a vital tie between cultural, social, and interpersonal processes. PMID- 10388946 TI - Motorcycles for the disabled: mobility, modernity and the transformation of experience in urban China. AB - This paper describes changes in people's attitudes toward and experiences of disability in contemporary China. In particular, it examines how, as a result of shifting gender structures and modernist modes of production, urban men who struggle to walk have adopted cycle technologies, and how this has caused Chinese society increasingly to associate these men with disability. The paper further details ways the young state-run advocacy organization, the China Disabled Persons' Federation, has contributed to these attitudinal and experiential shifts by providing more assistance to urban men who struggle to walk than to any other PRC citizens who might be considered disabled. In general, the transformations outlined in this paper exemplify how ongoing macro changes in contemporary China often provide benefits to a relatively small number of people and how, for those who receive them, the benefits are often double-edged. PMID- 10388947 TI - Cardiac pacemakers: current and future status. PMID- 10388948 TI - Green hair: causes and management. PMID- 10388949 TI - Botanical briefs: chrysanthemums--dendranthema spp. PMID- 10388950 TI - Melanoma precursors in children. PMID- 10388951 TI - Recurrent plantar keloid. AB - Keloids of the plantar foot present a unique challenge to the surgical dermatologist. Many of the established regimens often fall short of their desired goals. Some of the obstacles to overcome include the repetitive nature of ambulation, the inability to primarily close the plantar foot, and the exquisite tendency for even fine suturing of skin grafts to form keloids. The use of excision, postoperative electron beam therapy, and secondary intention healing provides a useful approach in the management of plantar keloids. PMID- 10388952 TI - Galvanic urticaria. AB - A variety of environmental stimuli, such as vibration, ultraviolet radiation, and exposure to water, are recognized as causes of "physical urticaria." A medical student, participating in a demonstration of a galvanic device used in the treatment of hyperhidrosis, demonstrated urticaria in response to this galvanic stimulation. PMID- 10388953 TI - Pretibial dystrophic epidermolysis bullosa. AB - Pretibial epidermolysis bullosa is a rare variant of dystrophic epidermolysis bullosa, characterized by bullae and violaceous lichenoid papules and plaques of the anterior aspects of the legs. A case of pretibial epidermolysis bullosa is presented and the literature is reviewed. PMID- 10388954 TI - Toxic epidermal necrolysis occurring as a consequence of treatment with foscarnet. AB - Toxic epidermal necrolysis (TEN) has been shown to occur following administration of many different medications. Recently, we observed a patient who sustained a severe case of TEN shortly following the administration of foscarnet. Since this agent has not been previously associated with this complication, we report the first case of TEN occurring secondary to treatment with foscarnet. PMID- 10388955 TI - Hereditary ochronosis: hyperpigmented skin overlying cartilaginous structures. AB - Hereditary ochronosis, or alkaptonuria, results from deficiency of homogentisic acid oxidase. It is an autosomal recessive condition found in geographically isolated populations. The excess homogentisic acid deposits in collagenous structures, leading to unusual pigmentation of the skin overlying cartilaginous structures, but on occasion pigment is also seen in the sclera, in sweat after oxidation, and classically, in urine when left standing at room temperature. This case report highlights the pathogenesis and expression of this rare disorder. PMID- 10388956 TI - Fatal cutaneous squamous cell carcinoma with extension through the maxillary sinus and orbit into the brain. AB - Cutaneous squamous cell carcinomas may cause death by metastasis or by local extension. We describe a deeply invasive cutaneous squamous cell carcinoma that caused death by direct extension into the brain. PMID- 10388957 TI - Detection and treatment of nonmelanoma skin cancer prevent disfigurement and death. PMID- 10388958 TI - Patients urged to seek treatment for actinic keratoses, recommends the American Academy of Dermatology, the American Cancer Society, and the Skin Cancer Foundation. PMID- 10388959 TI - Tazarotene gel is safe and effective in the treatment of acne vulgaris: a multicenter, double-blind, vehicle-controlled study. AB - Retinoids reverse the abnormal pattern of keratinization seen in acne vulgaris. Tazarotene is the first of a novel family of topical receptor-selective acetylenic retinoids. This study evaluates the safety and efficacy of topical tazarotene 0.1% and 0.05% gels, in comparison to vehicle gel, applied once daily for 12 weeks, in the treatment of mild-to-moderate facial acne vulgaris. A total of 446 patients with facial acne vulgaris were enrolled, and 375 patients, ranging in age from 14 to 44 years, were evaluable in this multicenter, double blind, randomized study. In comparison to vehicle gel, treatment with tazarotene 0.1% gel resulted in significantly greater reductions in noninflammatory and total lesion counts at all follow-up visits, and inflammatory lesion counts at Week 12. Tazarotene 0.05% gel resulted in significantly greater reductions in noninflammatory and total lesion counts than vehicle gel at Weeks 8 and 12. At Week 12, treatment success rates were 68% and 51% for tazarotene 0.1% and 0.05%, respectively (40% for vehicle gel). Tazarotene gel was an effective, safe, and generally well-tolerated therapy for the treatment of acne vulgaris. PMID- 10388960 TI - Is pregnancy in diabetic women associated with folate deficiency? AB - OBJECTIVE: To determine whether folate metabolism in pregnant diabetic women is significantly different from that in pregnant nondiabetic women, thus predisposing them to having offspring with major congenital anomalies. RESEARCH DESIGN AND METHODS: A total of 31 pregnant diabetic women and 54 pregnant nondiabetic control subjects were studied at their first prenatal visits. Dietary folate intake, serum folate, red blood cell folate, urinary folate, and homocysteine were measured and compared after controlling for folate supplementation. Among diabetic women the relationships among parameters of folate metabolism and glycemic control were also assessed. RESULTS: There were no significant differences between the pregnant diabetic and non-diabetic women for any measures of folate metabolism after accounting for folate supplementation. In addition, among diabetic women, there were no associations among parameters of folate metabolism and glycemic control. CONCLUSIONS: Abnormal folate metabolism does not appear to occur in pregnant diabetic women. It is unlikely that deranged folate metabolism explains the higher incidence of major anomalies in infants of diabetic mothers. These results do not diminish the importance of periconception folate supplementation or preclude other possible scenarios that could restrict folate use by the embryo, leading to congenital anomalies. PMID- 10388961 TI - Blood glucose awareness training and epinephrine responses to hypoglycemia during intensive treatment in type 1 diabetes. AB - OBJECTIVE: To determine the effect of blood glucose awareness training (BGAT) on epinephrine and symptom responses to hypoglycemia in patients with type 1 diabetes enrolled in an intensive diabetes treatment (IDT) program. RESEARCH DESIGN AND METHODS: A total of 47 subjects with uncomplicated diabetes (duration 9 +/- 3 years: HbA1c 9.0 +/- 1.2%; reference range 4-6%) enrolled in a 4-month outpatient IDT program were randomized to classes in BGAT (n = 25) (BGAT group) or cholesterol awareness (n = 22) (control group). Subjects underwent stepped hypoglycemic clamp studies before and at completion of IDT. Plasma glucose was lowered from 6.7 mmol/l (baseline) to 4.4, 3.9, 3.3, 2.8, and 2.2 mmol/l over 190 min. Symptoms, counterregulatory hormones, and ability of the subject to estimate their glucose level were assessed at each plateau. At home, subjects used a handheld computer to first estimate and then measure and record blood glucose levels for 70 trials over a 4-week period immediately before IDT and again immediately following the educational intervention. RESULTS: HbA1c decreased in both BGAT group (9.1 +/- 1.4 to 7.9 +/- 1.1%; P < 0.001) and control group (9.0 +/- 1.1 to 7.8 +/- 0.8%; P < 0.001) (NS between groups). Frequency of hypoglycemia (< 3.9 mmol/l) increased in both groups, from 0.45 +/- 0.06 to 0.69 +/- 0.07 episodes per day (P < 0.001) in the BGAT group and from 0.50 +/- 0.08 to 0.68 +/- 0.06 episodes per day (P < 0.05) in the control group NS between groups). Epinephrine responses after IDT were greater in the BGAT group (repeated measure analysis of variance [ANOVA], F = 3.5, P < 0.05). A separate analysis of subjects n = 26) most at risk for hypoglycemia (HbA1c after IDT < 7.8% or an HbA1c improvement of > 2 percentage points) showed that frequency of hypoglycemia increased in both the groups: from 0.50 +/- 0.09 to 0.80 +/- 0.11 episodes per day (P < 0.01) in the BGAT group (n = 14) and from 0.43 +/- 0.11 to 0.75 +/- 0.07 episodes per day (P < 0.05) in the control group (n = 12) (NS between groups). However, the epinephrine response in control subjects decreased with IDT while the response in the BGAT subjects was preserved (repeated measure ANOVA, F = 4.4, P < 0.02). CONCLUSIONS: BGAT is a useful intervention to decrease blunting of counterregulatory responses associated with improved glycemic control and may modify the severity of hypoglycemia associated with improved glycemic control in type 1 diabetes. PMID- 10388962 TI - Lower-extremity amputation in diabetes. The independent effects of peripheral vascular disease, sensory neuropathy, and foot ulcers. AB - OBJECTIVE: To identify risk factors for lower-extremity amputation (LEA) in individuals with diabetes and to estimate the incidence of LEA. RESEARCH DESIGN AND METHODS: This is a prospective study of 776 U.S. veterans in a general medicine clinic in Seattle, Washington. The outcome was first LEA during follow up. Potential risk factors evaluated in proportional hazards models included, among others, peripheral vascular disease (PVD), sensory neuropathy, former LEA, foot deformities and ulcers, diabetes duration and treatment, and hyperglycemia. RESULTS: Associated with an increased risk for LEA were PVD defined as transcutaneous oxygen < or = 50 mmHg (relative risk [RR] = 3.0, 95% CI 1.3-7.1), insensitivity to monofilament testing (RR = 2.9, odds ratio = 1.1-7.8), lower extremity ulcers (RR = 2.5, CI 1.1-5.4), former LEA, and treatment with insulin when controlling for duration of diabetes and other factors in the model. PVD defined as absent or diminished lower-extremity pulses or an ankle arm index < or = 0.8 was also associated with a significantly higher risk of LEA in separate models. Foot ulcers were associated with an increased ipsilateral risk of amputation. The age-adjusted incidence among men only for LEA standardized to the 1991 U.S. male diabetic population was 11.3/1,000 patient-years. CONCLUSIONS: This prospective study shows that peripheral sensory neuropathy, PVD, foot ulcers (particularly if they appear on the same side as the eventual LEA), former amputation, and treatment with insulin are independent risk factors for LEA in patients with diabetes. PMID- 10388963 TI - A prospective study of risk factors for diabetic foot ulcer. The Seattle Diabetic Foot Study. AB - OBJECTIVE: Little prospective research exists on risk factors for diabetic foot ulcer that considers the independent effects of multiple potential etiologic agents. We prospectively studied the effects of diabetes characteristics, foot deformity, behavioral factors, and neurovascular function on foot ulcer risk among 749 diabetic veterans with 1,483 lower limbs. RESEARCH DESIGN AND METHODS: Eligible subjects included all diabetic enrollees of a general internal medicine clinic without foot ulcer, of whom 83% agreed to participate. Baseline assessment included history and lower-limb physical examination, tests for sensory and autonomic neuropathy, and measurements of macro- and microvascular perfusion in the foot. Subjects were followed for the occurrence of a full thickness skin defect on the foot that took > 14 days to heal, with a mean follow-up of 3.7 years. RESULTS: Using stepwise Cox regression analysis, the following factors were independently related to foot ulcer risk: foot insensitivity to the 5.07 monofilament (relative risk [95% CI]) 2.2 (1.5-3.1), past history of amputation 2.8 (1.8-4.3) or foot ulcer 1.6 (1.2-2.3), insulin use 1.6 (1.1-2.2), Charcot deformity 3.5 (1.2-9.9), 15 mmHg higher dorsal foot transcutaneous PO2 0.8 (0.7 0.9), 20 kg higher body weight 1.2 (1.1-1.4), 0.3 higher ankle-arm index 0.8 (0.7 1.0), poor vision 1.9 (1.4-2.6), and 13 mmHg orthostatic blood pressure fall 1.2 (1.1-1.5). Higher ulcer risk was associated with hammer/claw toe deformity and history of laser photocoagulation in certain subgroups. Unrelated to foot ulcer risk in multivariate models were diabetes duration and type, race, smoking status, diabetes education, joint mobility, hallux blood pressure, and other foot deformities. CONCLUSIONS: Certain foot deformities, reduced skin oxygenation and foot perfusion, poor vision, greater body mass, and both sensory and autonomic neuropathy independently influence foot ulcer risk, thereby providing support for a multifactorial etiology for diabetic foot ulceration. PMID- 10388964 TI - Low birth weight, family history of diabetes, and glucose intolerance in Swedish middle-aged men. AB - OBJECTIVE: To investigate the association between low birth weight and glucose intolerance in relation to family history of diabetes. RESEARCH DESIGN AND METHODS: We conducted a population-based cross-sectional study of 2,237 men born in 1938-1957 in four municipalities in the outskirts of Stockholm, 50% of whom had a family history of diabetes (at least one first-degree or two second-degree relatives with diabetes). Oral glucose tolerance testing detected 35 cases of type 2 diabetes, 102 cases of impaired glucose tolerance, and 57 cases of impaired fasting glucose. RESULTS: In subjects without a family history of diabetes, low (< or = 3,000 g) birth weight was associated with an odds ratio of 2.3 (95% confidence intervals = 0.4-14.4) for diabetes, 1.8 (0.7-4.3) for impaired glucose tolerance, and 3.3 (1.0-10.4) for impaired fasting glucose. In subjects with a family history of diabetes, the corresponding figures were approximately similar, except for diabetes, for which the odds ratio was 5.4 (2.0 14.9). For men with low birth weight in combination with a family history of diabetes, the odds ratio was 10.9 (2.9-41.2) for diabetes, 2.4 (1.1-5.6) for impaired glucose tolerance, and 5.9 (2.1-16.3) for impaired fasting glucose. CONCLUSIONS: This study indicated that low birth weight is associated with type 2 diabetes, impaired glucose tolerance, and impaired fasting glucose in men. This finding was most pronounced in subjects with diabetes in the family, but it was also indicated in those without a family history of diabetes. Men with the combination of low birth weight and family history of diabetes seem to be at particularly high risk of developing type 2 diabetes. PMID- 10388965 TI - Prevalence of "syndrome X" features in parents of type 1 diabetic patients with or without nephropathy. AB - OBJECTIVE: To compare the prevalence of "syndrome X"-related parameters in parents of type 1 diabetic patients with diabetic nephropathy (DNP) to those in parents of diabetic patients without DNP. RESEARCH DESIGN AND METHODS: In this cross-sectional study, we included 50 parents of type 1 diabetic patients with DNP and 50 parents of diabetic patients without DNP. All parents were investigated in a fasting state for serum lipids including nonesterified fatty acids (NEFAs), glucose, HbA1c, plasma uric acid, fasting insulin levels, and albuminuria. Blood pressure was recorded in the supine position using an automatic device; ankle/brachial index was measured with Doppler ultrasound. Presence of cardiovascular disease was determined by a standardized questionnaire and electrocardiogram registration. Subjects without known diabetes underwent a 2 h oral glucose tolerance test. Anthropometric parameters such as BMI, waist-to hip ratio, and percentage of body fat were measured. In addition to univariate analysis, a syndrome X score (SXS) was formulated, comprising a number of syndrome X-related biochemical, physiological, and/or anthropometric parameters. RESULTS: Univariate analysis revealed no significant differences in syndrome X parameters between parents of type 1 diabetic patients with or without DNP. Also, the composite SXS was similar in both groups. CONCLUSIONS: In this study, no differences were found in the prevalence of syndrome X features between parents of type 1 diabetic patients with DNP and parents of patients without DNP. PMID- 10388966 TI - Early postpartum metabolic assessment in women with prior gestational diabetes. AB - OBJECTIVE: To present the results of early postpartum metabolic assessment in women with gestational diabetes mellitus (GDM), to determine predictive factors for subsequent diabetes, and to investigate the association of postpartum glucose tolerance with other components of the metabolic syndrome. RESEARCH DESIGN AND METHODS: A total of 788 women were evaluated 3-6 months after a GDM pregnancy. A 75-g oral glucose tolerance test (OGTT) was performed. Cholesterol, HDL cholesterol, triglycerides, blood pressure, BMI, and body fat distribution were assessed. Clinical and obstetric history, baseline variables at the diagnosis of GDM, metabolic control during pregnancy, and index pregnancy outcome were compared in women with diabetes and women without diabetes (American Diabetes Association [ADA] criteria) after pregnancy. Multivariate logistic regression analysis was used to ascertain independent predictors of subsequent diabetes. Correlation coefficients were assessed between postpartum glucose tolerance and lipid levels, blood pressure, BMI, and body fat distribution. RESULTS: According to ADA criteria, 588 (74.6%) women were normal, 46 (5.8%) had impaired fasting glucose, 82 (10.4%) had impaired glucose tolerance, 29 (3.7%) had both impaired fasting glucose and impaired glucose tolerance, and 43 (5.4%) had diabetes. Prepregnancy obesity, recurrence of GDM, gestational age at diagnosis of GDM, glucose values in the 100-g OGTT, number of abnormal values in the 100-g OGTT, fasting C-peptide levels in pregnancy, C-peptide/glucose score in pregnancy, insulin requirement in pregnancy, 3rd trimester HbA1c levels, and macrosomia differed significantly in women with subsequent diabetes. Independent predictors of postpartum diabetes were prepregnancy obesity, C-peptide/glucose score during pregnancy, and the number of abnormal values in the 100-g diagnostic OGTT. The area under the postpartum glucose curve was positively associated with BMI, waist circumference, waist-to-hip ratio, triglycerides, and systolic and diastolic blood pressures. CONCLUSIONS: Low C-peptide/glucose score during pregnancy together with prepregnancy obesity and severity of GDM (number of abnormal values in the 100-g diagnostic OGTT) are independent predictors of subsequent diabetes. Our data suggest that regardless of obesity and severity of GDM, a beta-cell defect increases the risk of postpartum diabetes. The association of postpartum glucose tolerance with triglyceride levels, blood pressure, obesity, and regional distribution of body fat suggests that postpartum glucose intolerance anticipates a high-risk cardiovascular profile that comprises other risk factors besides diabetes. PMID- 10388967 TI - Birth weight of women with gestational diabetes. AB - OBJECTIVE: Epidemiological observations have suggested a relationship between type 2 diabetes and a low birth weight. However, there are many confounding variables and problems with retrospective data collection. Women with gestational diabetes mellitus (GDM), who are likely to develop type 2 diabetes in the future, may help clarify these observations. RESEARCH DESIGN AND METHODS: Consecutive women with GDM (n = 138) were included in the study if they had a singleton pregnancy delivered between 37 and 41 weeks of gestation, if they had themselves been born in the local hospital, and if their own delivery data were available. With respect to their own births, a matched group was obtained by considering the next female delivery of the same gestational age. RESULTS: For women with GDM, the mean (+/- 1 SD) birth weight was 3,293 +/- 493 g and the ponderal index was 27.0 +/- 2.4. Their values were not significantly different from the matched group, which had a birth weight of 3,315 +/- 460 g and a ponderal index of 27.0 +/- 2.5. After adjusting for the gestational age of delivery, the birth weight of women with GDM did not show a U-shaped distribution. CONCLUSIONS: After adjustment for the gestational age of delivery, women with GDM do not themselves have either a lower or higher birth weight than a matched group. These data suggest that women with GDM are either not a good surrogate for investigating the relationship between birth weight and type 2 diabetes or that correction for the gestational age of delivery removes the most important confounding variable. It is also possible that modern dietary changes may have altered the relationship. PMID- 10388968 TI - Previous maternal abortion, longer gestation, and younger maternal age decrease the risk of type 1 diabetes among male offspring. AB - OBJECTIVE: To identify possible influences and interactions of perinatal determinants in the subsequent development of type 1 diabetes. RESEARCH DESIGN AND METHODS: The data were obtained from children born in Denmark during the periods 1978-1982 and 1984-1986 and admitted to a Danish hospital with newly diagnosed type 1 diabetes between 1978 and 1995; 857 patients fulfilled the criteria. The study was conducted by combining and analyzing two national registries: the National Patient Registry and the Medical Birth Registry. For each diabetic child, two control children were randomly selected, matched by sex, time, and district of delivery. RESULTS: By multivariate logistic regression analysis, the following significant determinants were identified. Male offspring showed decreased risk when born of mothers who had had one or more abortions (odds ratio [OR] 0.66 [95% CI 0.48-0.92]) and with long duration of gestation (linearly with OR 0.91 per week [0.85-0.99]), while increased risk was found for high maternal age (linearly with OR 1.03 per year [1.00-1.06]). Female offspring showed no such association. No significant differences between diabetic patients and control subjects were found with respect to paternal age, maternal parity, placental weight or any of the birth size parameters, or interventions and complications during delivery. CONCLUSIONS: The findings show that perinatal determinants may influence the risk of subsequent development of type 1 diabetes in a sex-specific manner. PMID- 10388969 TI - The onset age of type 1 diabetes in Finnish children has become younger. The Finnish Childhood Diabetes Registry Group. AB - OBJECTIVE: To analyze the change in the age distribution at onset of type 1 diabetes in boys and girls aged 1-14 years during a 32-year period (from 1965 to 1996). RESEARCH DESIGN AND METHODS: Data on the incidence of type 1 diabetes in Finland were obtained from the Central Drug Registry of the Social Insurance Institution for 1965-1986 (6,195 cases) and from the Prospective Childhood Diabetes Registry for 1987-1996 (3,613 cases). The change in age- and sex specific incidence was estimated by fitting the linear regression with the logarithm of the annual incidence data. Analysis of variance was used to compare the trends between the various age-groups (1-4, 5-9, and 10-14 years) and sexes. RESULTS: The incidence of type 1 diabetes increased predominantly in the younger age-groups. In children aged 1-4 years, the increase was 4.2% per year, and the overall 32-year relative increase was 338%. For children aged 5-9 and 10-14 years, the increase was 2.5 and 1.3% per year, respectively, and the overall relative increase was 116 and 49%, respectively. In boys aged 1-9 years, the increase was greatest from 1965 to 1984, whereas in girls aged 1-9 years, the statistically significant increase occurred between 1985 and 1996. In children aged 10-14 years, the only significant increase was seen in boys from 1965 to 1974 (3.7% per year). CONCLUSIONS: The greatest increase in the incidence of type 1 diabetes mainly occurred in children aged < 5 years. The incidence in young boys has been increasing since the mid-1960s, whereas in young girls, the significant increase began later, around the mid-1970s. In children aged 10-14 years, the increase in incidence has leveled off. PMID- 10388970 TI - Depressive symptoms and occurrence of type 2 diabetes among Japanese men. AB - OBJECTIVE: To examine the relationship between depressive symptoms and the incidence of type 2 diabetes. RESEARCH DESIGN AND METHODS: In 1984, 2,764 male employees of an electrical company in Japan completed a self-administered questionnaire including the Zung Self-Rating Depression Scale (SDS). They were followed for the next 8 years, and 2,380 (86%) responded to the follow-up survey in 1992. During the follow-up survey, occurrence of type 2 diabetes was diagnosed according to World Health Organization criteria. RESULTS: A total of 41 cases of type 2 diabetes were identified during the 8-year follow-up survey. After controlling for other known risk factors for type 2 diabetes, a proportional hazard analysis indicated that subjects who had moderate or severe levels of depressive symptoms (> or = 48 on the SDS) at baseline had a 2.3 times higher risk of having type 2 diabetes at the follow-up survey than those who were not depressed (< or = 39 on the SDS) (P < 0.05). CONCLUSIONS: Depressive symptoms may be associated with the onset of type 2 diabetes. PMID- 10388971 TI - Prospective associations of fasting insulin, body fat distribution, and diabetes with risk of ischemic stroke. The Atherosclerosis Risk in Communities (ARIC) Study Investigators. AB - OBJECTIVE: We tested the hypothesis that diabetes, body fat distribution, and (in nondiabetic subjects) fasting insulin levels are positively associated with ischemic stroke incidence in the general population. RESEARCH DESIGN AND METHODS: As part of the Atherosclerosis Risk in Communities (ARIC) Study, we measured diabetes by using fasting glucose criteria, waist and hip circumferences, and fasting insulin levels with a radioimmunoassay in > 12,000 adults aged 45-64 years who had no cardiovascular disease at baseline. We followed them for 6-8 years for ischemic stroke occurrence (n = 191). RESULTS: After adjustment for age, sex, race, ARIC community, smoking, and education level, the relative risk of ischemic stroke was 3.70 (95% CI 2.7-5.1) for diabetes, 1.74 (1.4-2.2) for a 0.11 increment of waist-to-hip ratio, and 1.19 (1.1-1.3) for a 50-pmol/l increment of fasting insulin among nondiabetic subjects. Ischemic stroke incidence was not statistically significantly associated with BMI (comparably adjusted relative risk = 1.15, 95% CI 0.97-1.36). With adjustment for other stroke risk factors (some of which may mediate the effects of diabetes, fat distribution, and hyperinsulinemia), the relative risks for diabetes, waist-to hip ratio, and fasting insulin level were 2.22 (95% CI 1.5-3.2), 1.08 (0.8-1.4), and 1.14 (1.01-1.3), respectively. CONCLUSIONS: Diabetes is a strong risk factor for ischemic stroke. Aspects of insulin resistance, as reflected by elevated waist-to-hip ratios and elevated fasting insulin levels, may also contribute to a greater risk of ischemic stroke. PMID- 10388973 TI - Is fasting leptin associated with insulin resistance among nondiabetic individuals? The Miami Community Health Study. AB - OBJECTIVE: Whether serum leptin levels are associated with insulin resistance independent of the effects of hyperinsulinemia and adiposity is an important unanswered question. We examined the relationship between the rate of insulin mediated glucose uptake and serum leptin concentrations among nondiabetic men and women. RESEARCH DESIGN AND METHODS: A cross-sectional analysis was performed among 49 young to middle-aged men and women who participated in the Miami Community Health Study. All participants had measures of insulin resistance (euglycemic-hyperinsulinemic clamp), postchallenge insulin levels, fasting serum leptin levels, and several measures of adiposity. RESULTS: The rate of insulin mediated glucose uptake (M in milligrams per kilogram per minute) was significantly associated with leptin concentrations in both men (r = -0.83; P < 0.001) and women (r = -0.59; P < 0.001). M was also inversely related to percent body fat and to the 2-h insulin area under the curve (AUC). After covariate adjustment for sex, percent body fat, and AUC, leptin remained a significant correlate of M (P = 0.04). CONCLUSIONS: Cross-sectionally, leptin was significantly associated with insulin resistance in this nondiabetic sample of men and women. There may be a different physiological mechanism to explain the leptin/insulin resistance association apart from the insulin/adiposity link. Confirmatory evidence awaits the results of clinical trials. PMID- 10388974 TI - Leptin and variables of body adiposity, energy balance, and insulin resistance in a population-based study. The Hoorn Study. AB - OBJECTIVE: Leptin is thought to play a key role in the control of body weight. There is a complex interrelationship between leptin and insulin or insulin resistance, but it is unknown how leptin is regulated. We therefore explored, in a large population-based study of 2,484 Caucasian subjects aged 50-74 years, the relationship between leptin and variables of body adiposity, energy balance, and insulin resistance. RESEARCH DESIGN AND METHODS: Leptin was measured by means of a radioimmunoassay. Multiple linear regression analyses were performed with leptin as dependent variable and age, sex, BMI, waist circumference, daily energy intake, physical activity, smoking, hypertension, fasting triglyceride concentrations, HDL cholesterol, fasting plasma glucose, and fasting plasma insulin concentrations as independent variables (determinants) RESULTS: Leptin concentrations were found to be four times higher in women than in men. Effect modification between sex and potential determinants was expected, and the analyses were performed separately for women and men. BMI was the strongest determinant of leptin in women and waist circumference the strongest determinant in men. BMI, waist circumference, insulin, and triglyceride concentrations were independently and significantly (P < 0.05) associated with leptin, while inverse associations were shown for smoking and daily energy intake (borderline significance). CONCLUSIONS: This study confirms the relationship between insulin and leptin and, in addition, suggests a relationship between triglyceride concentrations and leptin independent of sex, BMI, waist circumference, and insulin. PMID- 10388972 TI - Improved glycemic control reduces the impact of weight gain on cardiovascular risk factors in type 1 diabetes. The Epidemiology of Diabetes Complications Study. AB - OBJECTIVE: To assess the prevalence and incidence of being overweight in type 1 diabetes, to identify factors associated with weight gain and improved glycemic control, and to examine relationships among weight gain, glycemic control, and cardiovascular risk factors. RESEARCH DESIGN AND METHODS: The prevalence and incidence of being overweight in the Pittsburgh Epidemiology of Diabetes Complications (EDC) cohort (n = 441) were compared with the general population (National Health and Nutrition Examination Survey [NHANES]). Factors associated with weight gain and improved glycemic control were identified, and relationships among weight gain, glycemic control, and cardiovascular risk factors were examined over a 6.9 +/- 2.2-year period. RESULTS: At baseline, the prevalence of being overweight (BMI > 27.8 kg/m2 for men and > 27.3 kg/m2 for women) was 10.4 and 11.4%, respectively, and was lower than the age- and sex-specific estimate for the general population (P < 0.05). The incidence of being overweight was comparable in men (12.6%) and women (11.8%) and did not differ from the general population (P = 0.98). Weight gain correlated with improvements in HbA1c (r = 0.21, P < 0.001). Patients with the highest baseline HbA1c levels gained the most weight and had the greatest improvement in glycemic control. A lower baseline BMI was also associated with a greater improvement in glycemic control. Weight gain favorably influenced the lipid profile in the setting of improved glycemic control, but adversely influenced the lipid profile in the absence of improved glycemic control. Weight change was directly associated with blood pressure change, but the incidence of hypertension was more strongly influenced by the development of nephropathy. CONCLUSIONS: The prevalence of being overweight in type 1 diabetes remains lower than that in the general population. Moderate weight gain did not adversely affect the cardiovascular risk profile in the setting of improved glycemic control. PMID- 10388975 TI - Work disability and diabetes. AB - OBJECTIVE: To determine the rates and demographic determinants of work disability, hours worked per week, work-loss days, and wages in individuals with diabetes. RESEARCH DESIGN AND METHODS: A probit regression analysis was performed on a cross-sectional population-based survey of U.S. noninstitutionalized civilian population (National Medical Expenditures Survey--2, 1987). The sample was restricted to individuals aged > or = 25 years. A total of 1,502 individuals reported having diabetes, and 20,405 did not. Information on workforce participation and income were collected quarterly. Work disability was defined as a self-report of having been unable to work because of illness or disability for > or = 2 quarters in 1987. RESULTS: Work disability was reported by 25.6% of individuals with diabetes, compared with 7.8% of those without diabetes. Work disability rates were higher for older people, females, and African-Americans, and lower for Hispanics and for individuals with greater non-wage income. Individuals with diabetes engaged in the workforce had more work-loss days than did nondiabetic individuals, but had similar hourly wages. Predicted mean earnings were significantly lower for individuals with diabetes at all ages, resulting in $4.7 million loss in earnings in 1987 due to work disability. CONCLUSIONS: Work disability is significantly higher for individuals with diabetes than for those without diabetes at all ages, and results in a significant decrease in earnings. A disproportionate burden of work disability is borne by older individuals and women with diabetes. Better information on the determinants of work disability in individuals with diabetes is needed. PMID- 10388976 TI - Determining an episode of care using claims data. Diabetic foot ulcer. AB - OBJECTIVE: Amid changes in the organization and financing of health care, health care decision makers are increasingly interested in episodes of care. We sought to determine an episode of care for diabetic foot ulcer using an administrative claims database. RESEARCH DESIGN AND METHODS: We used 1993-1995 claims data to assess resource utilization for privately insured patients with diabetic foot ulcers. Over a 26-week period, we determined the episode length by comparing differences in average daily charges and proportion of patients with charges before and after foot ulcer diagnosis. All 13 weeks before diagnosis were used to calculate baseline values. Significance was determined by CIs, which were calculated by a nonparametric bootstrap technique. Costs associated with the episode were also calculated. A sensitivity analysis using weeks with highest and lowest values as baseline was also conducted. RESULTS: Based on average daily charges, the episode of care for diabetic foot ulcer was 5 weeks. Using proportion of patients with charges, the episode was longer than 13 weeks. The cost for an episode of care ranged from $900 to $2,600. In the sensitivity analyses, episodes of care ranged from 1 to 13 weeks. CONCLUSIONS: Episodes of care can be defined by the period beginning with increased resource consumption and ending when resource consumption returns to baseline levels. With the growth of managed care and disease management programs, episode-of-care analysis may have an increasingly important role in health care provision and delivery. PMID- 10388977 TI - Type 2 diabetes: incremental medical care costs during the first 8 years after diagnosis. AB - OBJECTIVE: To describe and analyze the time course of medical care costs caused by type 2 diabetes, from the time of diagnosis through the first 8 postdiagnostic years. RESEARCH DESIGN AND METHODS: From electronic health maintenance organization (HMO) records, we ascertained the ongoing medical care costs for all members with type 2 diabetes who were newly diagnosed between 1988 and 1995. To isolate incremental costs (costs caused by the diagnosis of diabetes), we subtracted the costs of individually matched HMO members without diabetes from costs of members with diabetes. RESULTS: The economic burden of diabetes is immediately apparent from the time of diagnosis. In year 1, total medical costs were 2.1 times higher for patients with diabetes compared with those without diabetes. Diabetes-associated incremental costs (type 2 diabetic costs minus matched costs for people without diabetes) averaged $2,257 per type 2 diabetic patient per year during the first 8 postdiagnostic years. Annual incremental costs varied relatively little over the period but were higher during years 1, 7, and 8 because of higher-cost hospitalizations for causes other than diabetes or its complications. CONCLUSIONS: For the first 8 years after diabetes diagnosis, patients with type 2 diabetes incurred substantially higher costs than matched nondiabetic patients, but those high costs remained largely flat. Once the growth in costs due to general aging is controlled for, it appears that diabetic complications do not increase incremental costs as early as is commonly believed. Additional research is needed to better understand how diabetes and its diagnosis affect medical care costs over longer periods of time. PMID- 10388978 TI - Quality of life in type 2 diabetic patients is affected by complications but not by intensive policies to improve blood glucose or blood pressure control (UKPDS 37). U.K. Prospective Diabetes Study Group. AB - OBJECTIVE: To determine in patients with type 2 diabetes the effects on quality of life (QOL) of therapies for improving blood glucose control and for improving blood pressure (BP) control, diabetic complications, and hypoglycemic episodes. RESEARCH DESIGN AND METHODS: We performed two cross-sectional studies of patients enrolled in randomized controlled trials of 1) an intensive blood glucose control policy compared with a conventional blood glucose control policy, and 2) a tight BP control policy compared with a less tight BP control policy. Also undertaken was a longitudinal study of patients in a randomized controlled trial of an intensive blood glucose control policy compared with a conventional blood glucose control policy. Subjects' QOL was assessed before or at the time of randomization and from 6 months to 6 years after randomization. Two cross-sectional samples of type 2 diabetic patients were randomized to therapies for blood glucose control: 1) 2,431 patients, mean age 60, duration from randomization 8.0 years, completed a "specific" questionnaire covering four aspects of QOL, and 2) 3,104 patients, mean age 62, duration from randomization 11 years, completed a "generic" QOL measure. Of these samples, 628 and 747 patients, respectively, were also randomized to therapies for BP control. A sample of 122 non-diabetic control subjects, average age 62, were also given the specific questionnaire. A longitudinal sample of 374 type 2 diabetic patients randomized to either intensive or conventional blood glucose policies, mean age at randomization 52, were given the specific questionnaire. Sample-sizes at 6 months and 1, 2, 3, 4, 5, and 6 years after randomization were 322, 307, 280, 253, 225, 163, and 184, respectively. The specific questionnaire assessed specific domains of QOL, including mood disturbance (Profile of Mood State), cognitive mistakes (Cognitive Failures Questionnaire), symptoms, and work satisfaction; the generic questionnaire (EQ5D) assessed general health. Both questionnaires were self administered. RESULTS: The cross-sectional studies showed that allocated therapies were neutral in effect, with neither improvement nor deterioration in QOL scores for mood, cognitive mistakes, symptoms, work satisfaction, or general health. The longitudinal study also showed no difference in QOL scores for the specific domains assessed, other than showing marginally more symptoms in patients allocated to conventional than to intensive policy. In the cross sectional studies, patients who had had a macrovascular complication in the last year had worse general health, as measured by the generic questionnaire, than those without complications, with scale scores median 60 and 78 respectively (P = 0.0006) and tariff scores median 0.73 and 0.83 respectively (P = 0.0012); more problems with mobility, 64 and 36%, respectively (P < 0.0001); and more problems with usual activities, 48 and 28% respectively (P = 0.0023). As measured by the specific questionnaire, they also showed reduced vigor (P = 0.0077). Patients who had had a microvascular complication in the last year reported more tension (P = 0.0082) and total mood disturbance (P = 0.0054), as measured by the specific questionnaire, than patients without complications. Patients treated with insulin who had had two or more hypoglycemic episodes during the previous year reported more tension (P = 0.0023), more overall mood disturbance (P = 0.0009), and less work satisfaction (P = 0.0042), as measured by the specific questionnaire, than those with no hypoglycemic attacks, after adjusting for age, duration from randomization, systolic BP, HbA1c, and sex in a multivariate polychotomous regression. CONCLUSIONS: In patients with type 2 diabetes, complications of the disease affected QOL, whereas therapeutic policies shown to reduce the risk of complications had no effect on QOL. It cannot be discerned whether frequent hypoglycemic episodes affect QOL, or whether patients with certain p PMID- 10388979 TI - Continuous subcutaneous infusion of glucagon-like peptide 1 lowers plasma glucose and reduces appetite in type 2 diabetic patients. AB - OBJECTIVE: The gut hormone glucagon-like peptide 1 (GLP-1) has insulinotropic and anorectic effects during intravenous infusion and has been proposed as a new treatment for type 2 diabetes and obesity. The effect of a single subcutaneous injection is brief because of rapid degradation. We therefore sought to evaluate the effect of infusion of GLP-1 for 48 h in patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: We infused GLP-1 (2.4 pmol.kg-1.min-1) or saline subcutaneously for 48 h in randomized order in six patients with type 2 diabetes to evaluate the effect on appetite during fixed energy intake and on plasma glucose, insulin, glucagon, postprandial lipidemia, blood pressure, heart rate, and basal metabolic rate. RESULTS: The infusion resulted in elevations of the plasma concentrations of intact GLP-1 similar to those observed after intravenous infusion of 1.2 pmol.kg-1.min-1, previously shown to lower blood glucose effectively in type 2 diabetic patients. Fasting plasma glucose (day 2) decreased from 14.1 +/- 0.9 (saline) to 12.2 +/- 0.7 mmol/l (GLP-1), P = 0.009, and 24-h mean plasma glucose decreased from 15.4 +/- 1.0 to 13.0 +/- 1.0 mmol/l, P = 0.0009. Fasting and total area under the curve for insulin and C-peptide levels were significantly higher during the GLP-1 administration, whereas glucagon levels were unchanged. Neither triglycerides nor free fatty acids were affected. GLP-1 administration decreased hunger and prospective food intake and increased satiety, whereas fullness was unaffected. No side effects during GLP-1 infusion were recorded except for a brief cutaneous reaction. Basal metabolic rate and heart rate did not change significantly during GLP-1 administration. Both systolic and diastolic blood pressure tended to be lower during the GLP-1 infusion. CONCLUSIONS: We conclude that 48-h continuous subcutaneous infusion of GLP-1 in type 2 diabetic patients 1) lowers fasting as well as meal-related plasma glucose, 2) reduces appetite, 3) has no gastrointestinal side effects, and 4) has no negative effect on blood pressure. PMID- 10388980 TI - Counterregulation during spontaneous nocturnal hypoglycemia in prepubertal children with type 1 diabetes. AB - OBJECTIVE: To examine counterregulatory responses during spontaneous nocturnal hypoglycemia in prepubertal children with type 1 diabetes. RESEARCH DESIGN AND METHODS: A total of 29 prepubertal patients with type 1 diabetes underwent two overnight profiles. Data were analyzed from 16 children (median [range] 8.7 [5.9 12.9] years of age) with a night of hypoglycemia and a nonhypoglycemic night. Children hypoglycemic (< 3.5 mmol/l) on night 1 were given 25% extra carbohydrate as uncooked cornstarch with their usual evening snack on night 2 to avoid hypoglycemia. Glucose, growth hormone, and cortisol were measured every 15 min, catecholamines every 30 min, and glucagon, pancreatic polypeptide, insulin, and ketones every 60 min. A group of 15 healthy control subjects, aged 9.5 (5.6-12.1) years, underwent one overnight profile. RESULTS: Median duration of hypoglycemia was 225 (30-630) min, and glucose nadir was 2.0 (1.2-3.3) mmol/l. Insulin levels were not different on the two nights (P = 0.9, analysis of variance), but children with diabetes had higher insulin levels than normal control subjects between 2300 and 0300, maximal at 0200 (mean +/- SEM 57.4 +/- 5.7 vs. 31.6 +/- 5.0 pmol/l, P = 0.002). Peak epinephrine was higher on the night of hypoglycemia (0.98 [0.52-2.09] nmol/l) versus nonhypoglycemia (0.32 [0.21-0.62] nmol/l), P = 0.001, but norepinephrine (1.29 [1.07-2.64] vs. 1.26 [1.04-1.88] nmol/l, P = 0.5), glucagon (93 [64.2-125.6] vs. 100.5 [54.6-158] ng/l, P = 0.6), pancreatic polypeptide (410.2 [191-643.2] vs. 270.8 [158.2-777.8] ng/l, P = 0.5), and cortisol (513 [300-679] vs. 475 [235-739] nmol/l, P = 0.6) were not different. Glucose threshold for epinephrine release was very low, 1.9 +/- 0.2 mmol/l. There was a short-lived rise in growth hormone from 75-105 min after onset of hypoglycemia, maximal at 90 min (7.8 +/- 1.2 vs. 3.5 +/- 0.9 ng/ml, P = 0.02). CONCLUSIONS: The prolonged nature of nocturnal hypoglycemic episodes may be explained in part by defective counterregulation. The risk of nocturnal hypoglycemia needs to be reduced before intensification of insulin therapy can be contemplated in this age-group. PMID- 10388981 TI - Circadian blood pressure during the early course of type 1 diabetes. Analysis of 1,011 ambulatory blood pressure recordings in 354 adolescents and young adults. AB - OBJECTIVE: Little information is available on the early course of hypertension in type 1 diabetes. The aim of our study, therefore, was to document circadian blood pressure profiles in patients with a diabetes duration of up to 20 years and relate daytime and nighttime blood pressure to duration of diabetes, BMI, insulin therapy, and HbA1c. RESEARCH DESIGN AND METHODS: Ambulatory profiles of 24-h blood pressure were recorded in 354 pediatric patients with type 1 diabetes (age 14.6 +/- 4.2 years, duration of diabetes 5.6 +/- 5.0 years, follow-up for up to 9 years). A total of 1,011 profiles were available for analysis from patients not receiving antihypertensive medication. RESULTS: Although daytime mean systolic pressure was significantly elevated in diabetic subjects (+3.1 mmHg; P < 0.0001), daytime diastolic pressure was not different from from the height- and sex adjusted normal range (+0.1 mmHg, NS). In contrast, both systolic and diastolic nighttime values were clearly elevated (+7.2 and +4.2 mmHg; P < 0.0001), and nocturnal dipping was reduced (P < 0.0001). Systolic blood pressure was related to overweight in all patients, while diastolic blood pressure was related to metabolic control in young adults. Blood pressure variability was significantly lower in girls compared with boys (P < 0.01). During follow-up, no increase of blood pressure was noted; however, diastolic nocturnal dipping decreased significantly (P < 0.03). Mean daytime blood pressure was significantly related to office blood pressure (r = +0.54 for systolic and r = +0.40 for diastolic pressure); however, hypertension was confirmed by ambulatory blood pressure measurement in only 32% of patients with elevated office blood pressure. CONCLUSIONS: During the early course of type 1 diabetes, daytime blood pressure is higher compared with that of healthy control subjects. The elevation of nocturnal values is even more pronounced and nocturnal dipping is reduced. The frequency of white-coat hypertension is high among adolescents with diabetes, and ambulatory blood pressure monitoring avoids unnecessary antihypertensive treatment. PMID- 10388982 TI - Increased Na+/Li+ countertransport activity may help to identify type 1 diabetic adolescents and young adults at risk for developing persistent microalbuminuria. AB - OBJECTIVE: To evaluate whether erythrocyte sodium-lithium countertransport (Na+/Li+ CT) activity may identify adolescents and young adults with childhood onset of type 1 diabetes to be at greater risk to develop persistent microalbuminuria and incipient diabetic nephropathy. RESEARCH DESIGN AND METHODS: In January 1989, Na+/Li+ CT was measured in 170 normoalbuminuric diabetic adolescents and young adults (age 12-23 years; onset of diabetes before age 18 years; duration of diabetes longer than 7 years). Participants were clinically examined at baseline and biennially thereafter. Na+/Li+ CT activity was measured every 2 years during the 8-year follow-up period. Na+/Li+ CT activity was measured also in parents of diabetic offspring. RESULTS: Over 8 years, 18 (10 male, 8 female) out of 170 patients (10.5%) developed persistent microalbuminuria; no patient developed overt nephropathy. The risk of developing microalbuminuria was higher in children with increased Na+/Li+ CT (using 300 mumol.1 erythrocytes-1.h-1 as the arbitrary cutoff point) (group 1) compared with those with normal Na+/Li+ CT at the beginning of the study (group 2) (18.98 vs. 3.29%, P < 0.01; sensitivity 96.7%; specificity 57.9%). Sex did not influence predictive value, sensitivity, or specificity. Na+/Li+ CT was not significantly correlated with HbA1c or duration of type 1 diabetes. The percentage of offspring with both parents having Na+/Li+ CT activity above the median values was significantly higher in patients in group 1 than in group 2. The odds ratio for the occurrence of microalbuminuria after adjustment for confounding variables (albumin excretion rate [AER], sex, HbA1c, mean blood pressure, cholesterol, triglycerides) in type 1 diabetic adolescents with elevated baseline erythrocyte Na+/Li+ CT was 4.5 (95% CI of 2.1-11.4). CONCLUSIONS: These results confirm those of previous studies and suggest that Na+/Li+ CT may be one of the predictors and risk factors for incipient diabetic nephropathy in adolescents and young adults with onset of diabetes during childhood. Persistently increased Na+/Li+ CT activity may help to identify normotensive, normoalbuminuric patients with type 1 diabetes who are predisposed to develop microalbuminuria and incipient diabetic nephropathy. PMID- 10388984 TI - Effect of hyperketonemia on plasma lipid peroxidation levels in diabetic patients. AB - OBJECTIVE: This study was undertaken to examine the effect of ketosis on plasma lipid peroxidation levels in diabetic patients. RESEARCH DESIGN AND METHODS: Plasma levels of lipid peroxidation products (malondialdehyde) and ketone bodies (acetoacetate and beta-hydroxybutyrate) were determined in diabetic patients (n = 70) and age-matched normal volunteers (n = 25). Diabetic patients with total ketone body levels > 1.0 mmol/l were considered hyperketonemic, and those with levels < or = 1.0 mmol/l were considered normoketonemic. RESULTS: After normalization versus total lipids, levels of lipid peroxidation were significantly higher in the plasma of hyperketonemic diabetic patients (P < 0.05), but not in normoketonemic diabetic patients, compared with age-matched normal volunteers. In addition, low ketonemia was associated with lower lipid peroxidation levels when lipid peroxidation and ketonemia were determined in the same patient (n = 7) at two different clinic visits. CONCLUSIONS: This study demonstrated an association between hyperketonemia and increased lipid peroxidation levels in diabetic patients, which suggests that ketosis is a risk factor in the elevated lipid peroxidation levels associated with diabetes. Further investigation is needed to determine whether antioxidant supplementation can be particularly beneficial in reducing lipid peroxidation and complications in type 1 diabetic patients who frequently encounter ketosis. PMID- 10388983 TI - Increased small dense LDL and intermediate-density lipoprotein with albuminuria in type 1 diabetes. AB - OBJECTIVE: This population study examines the relationship between LDL density and persistent albuminuria in subjects with type 1 diabetes at the end of the Diabetes Control and Complications Trial (DCCT). RESEARCH DESIGN AND METHODS: Subjects were classified as persistently normoalbuminuric (albumin excretion rate [AER] < 30 mg/d, n = 1,056), microalbuminuric (AER > or = 30-299 mg/day, n = 80), and macroalbuminuric (AER = 300 mg/day, n = 24) based on the last two AER measures. RESULTS: Triglyceride (P < 0.01) and LDL cholesterol (P < 0.01) levels were higher in macroalbuminuric subjects compared with normoalbuminuric subjects. Cholesterol distribution by density-gradient ultracentrifugation showed an increase in intermediate-density lipoprotein (IDL) and a shift in peak LDL from buoyant toward more dense particles with progressive albuminuria. In the entire group, there was a significant negative correlation between the peak buoyancy of LDL particles and albuminuria (r = -0.238, P < 0.001, n = 1,160). This correlation persisted in the normoalbuminuric DCCT group (r = -0.138, P < 0.001, n = 1,056). CONCLUSIONS: As albuminuria increases in subjects with type 1 diabetes, dyslipidemia occurs with an increase in IDL and dense LDL that may lead to increased cardiovascular disease. PMID- 10388985 TI - Increased urinary transferrin excretion predicts microalbuminuria in patients with type 2 diabetes. AB - OBJECTIVE: We studied whether increased urinary transferrin excretion rates (TERs) (urinary transferrin-to-urinary creatinine ratio > or = 107 micrograms/mmol, which is the sum of an average and 2 SDs in 431 healthy nondiabetic individuals) would predict the development of microalbuminuria (urinary albumin-to-urinary creatinine ratio > or = 2.8 mg/mmol) in patients with type 2 diabetes and normal urinary albumin excretion rates (AERs) (albumin-to creatinine ratio < 2.8 mg/mmol). We also studied the influence of blood pressure, glycemic control, and serum levels of lipids and apolipoproteins on the later development of microalbuminuria. RESEARCH DESIGN AND METHODS: In 77 diabetic patients with normal AER, AER and TER were measured at baseline and after 24 months of follow-up. Blood pressure, glycemic control, and serum levels of lipids and apolipoproteins were measured at 1- to 2-month intervals during the follow-up period. RESULTS: Of the 16 patients who initially had increased TER, 5 (31%) developed microalbuminuria. In contrast, of the 61 who initially had normal TER, 4 (7%) developed microalbuminuria (P = 0.016). At baseline, no difference was found in age, sex, diabetes duration, diabetic medications, prevalence of hypertension, blood pressure, HbA1c levels, or serum lipid and apolipoprotein concentrations between the two group of patients with normal and increased TER. There was also no difference in duration of hypertension and prevalence of users of ACE inhibitors between two subgroups of hypertensive patients with normal and increased TER. During the 24 month follow-up period, those whose condition progressed to microalbuminuria had increased serum levels of triglycerides (1.87 +/- 0.49 vs. 1.29 +/- 0.64 mmol/l, P = 0.003) and apolipoprotein B (114 +/- 20 vs. 102 +/- 24 mg/dl, P = 0.05) and tended to have increased HbA1c levels (7.7 +/ 1.0 vs. 7.1 +/- 1.1%, P = 0.10) compared with those in whom microalbuminuria did not develop. Blood pressure, however, did not differ. In multivariate stepwise logistic regression analysis, the association between increased TER at baseline and subsequent development of microalbuminuria was significant (odds ratio 7.04 [95% CI 1.02-48.5], P = 0.04). CONCLUSIONS: In patients with type 2 diabetes and normal AER, increased TER may predict the development of microalbuminuria and abnormalities in triglyceride-rich lipoprotein metabolism, and poor glycemic control may be associated with this progression. PMID- 10388986 TI - Increased urinary atrial natriuretic peptide-like immunoreactivity excretion but decreased plasma atrial natriuretic peptide concentration in patients with hyperosmolar-hyperglycemic nonketotic syndrome. AB - OBJECTIVE: This study was undertaken to measure urinary atrial natriuretic peptide-like immunoreactivity (ANP-LI) and plasma ANP concentration in patients with hyperosmolar-hyperglycemic nonketotic syndrome (HHNS) to investigate the change of renal ANP-LI and cardiac ANP synthesis in volume-depleted diabetic patients. RESEARCH DESIGN AND METHODS: The urine ANP-LI:creatinine ratio, plasma ANP level, and plasma renin activity (PRA) were measured in 12 patients with HHNS during the acute stage and after recovery, in 28 oral hypoglycemic agent (OHA) treated type 2 diabetic patients, and in 23 normal subjects. ANP and PRA were measured by radioimmunoassay. RESULTS: These HHNS patients had severe hyperglycemia and hyperosmolality as well as increased blood urea nitrogen, creatinine, and PRA levels, as compared with normal subjects and OHA-treated type 2 diabetic patients. In these patients, the urinary ANP-LI:creatinine ratio (11.69 +/- 2.11 pmol/mmol) was significantly increased in comparison with the normal group (1.78 +/- 0.11 pmol/mmol) and OHA-treated diabetic patients (2.43 +/ 0.45 pmol/mmol), whereas plasma ANP concentration (5.12 +/- 0.72 pmol/l) was significantly lower than the corresponding values of the normal group (7.39 +/- 0.85 pmol/l) and OHA-treated diabetic patients (8.43 +/- 1.05 pmol/l). All of these abnormalities were significantly ameliorated after insulin, fluid, and electrolyte replacement. CONCLUSIONS: Our data show that urinary ANP-LI was significantly increased, whereas plasma ANP concentration was decreased, in the face of raised PRA in HHNS patients. This study indicates that renal ANP-LI substances and cardiac ANP may exhibit different responsiveness in diabetic patients with HHNS. PMID- 10388987 TI - Serum levels of advanced glycation end products are associated with left ventricular diastolic function in patients with type 1 diabetes. AB - OBJECTIVE: Impairment of left ventricular diastolic function, possibly caused by increased collagen cross-linking of the cardiac muscle, is common in patients with type 1 diabetes even without coronary artery disease. Advanced glycation end products (AGEs) cross-link tissue collagen and are found within myocardial fibers. The aim of this study was to examine for a possible association between circulating AGEs and left ventricular cardiac function. RESEARCH DESIGN AND METHODS: Left ventricular diastolic and systolic function were assessed by M-mode and Doppler echocardiography in 52 patients with type 1 diabetes, age 40 +/- 13 (mean +/- SD) years, diabetes duration 17 +/- 13 years, and HbA1c 8.3 +/- 1.1%. Serum levels of AGEs and N epsilon-(carboxymethyl)lysine (CML) were measured by newly developed competitive immunoassays. RESULTS: A positive correlation was found between serum levels of AGEs and isovolumetric relaxation time (IVRT), r = 0.46 (P < 0.0008), and left ventricular diameter during diastole, r = 0.37 (P < 0.008). The systolic parameters did not correlate with serum levels of AGEs. Stepwise regression analysis showed that 21% of the IVRT variation could be explained by serum levels of AGEs (F = 11.4, P < 0.002), whereas serum levels of CML, HbA1c, albumin excretion rate, diabetes duration, and mean arterial blood pressure were of no importance. AGE levels were significantly increased in men compared with women (P < 0.03) and present or former smokers (P < 0.04). CONCLUSIONS: Increased serum levels of AGEs, unlike serum levels of CML, are associated with heart stiffness in patients with type 1 diabetes, possibly mediated by the cross-linking properties of AGEs. PMID- 10388988 TI - Insulin resistance and classic risk factors in type 2 diabetic patients with different subtypes of ischemic stroke. AB - OBJECTIVE: In addition to classic risk factors (e.g., hypertension), insulin resistance is an important risk factor for the development of atherosclerosis. To investigate the risk factors for ischemic stroke in type 2 diabetes, we measured insulin sensitivity and several risk factors in 94 Japanese type 2 diabetic patients with different types of stroke. RESEARCH DESIGN AND METHODS: Stroke was classified by magnetic resonance imaging (MRI) and magnetic resonance (MR) angiography into the following subtypes: 1) patients with normal MRI and MR angiography (NOR; n = 30), 2) patients with lacunar infarction (LAC; n = 28), 3) patients with atherothrombotic infarction (ATI; n = 22), and 4) patients with large-artery atherosclerosis (LAA; n = 14). Insulin sensitivity was assessed by the K index of the insulin tolerance test (KITT). RESULTS: Patients with LAC, ATI, and LAA were significantly older and were more likely to be hypertensive than patients with NOR. Significantly higher insulin resistance was observed in patients with LAC, ATI, and LAA than in patients with NOR (KITT 2.21 +/- 0.17, 2.10 +/- 0.17, 2.19 +/- 0.25, and 3.25 +/- 0.21% per min, respectively, P < 0.001). Adjustment for age, sex, BMI, and duration of diabetes did not influence this result. Multiple logistical regression analysis showed that insulin resistance was an independent risk factor for all subtypes of ischemic stroke in type 2 diabetic patients. The same analysis showed that a high pulse pressure was a risk factor for LAC, postprandial C-peptide (hyperinsulinemia) was a risk factor for ATI, and longstanding hyperglycemia was a risk factor for LAA. PMID- 10388990 TI - What is the significance of macrosomia? AB - This commentary/review briefly considers the diverse criteria recommended for classification of overweight infants. Macrosomia continues to be a vexing problem for both obstetricians and pediatricians. Among the various techniques possible for use in assessing body composition, none are more practical than body weight relative to gestational age. The criteria for normative data from large populations are reviewed. The stringent definition, i.e., exceeding +2 SD of an appropriate normative population, is reaffirmed. Using these criteria, infants of diabetic mothers showed a significant relationship of body weight to fetal hyperinsulinemia. PMID- 10388989 TI - Bone mineral density in patients with type 1 and type 2 diabetes. AB - OBJECTIVE: To assess the effect of type 1 and type 2 diabetes and insulin treatment on bone mineral density (BMD) in middle-aged and elderly men and women. RESEARCH DESIGN AND METHODS: We measured BMD and evaluated known determinants of osteoporosis in 56 type 1 and 68 type 2 diabetic patients and 498 nondiabetic community control subjects. All patients, aged 52-72 years, developed diabetes after the age of 30 years (i.e., after achievement of peak bone mass) and were treated with insulin. BMD was measured at the proximal femur with dual-energy X ray absorptiometry. RESULTS: Among both sexes, BMD values were significantly lower in type 1 diabetic patients than in type 2 diabetic patients or the control subjects. When adjusted for age and BMI, the differences between type 1 diabetic patients and control subjects remained essentially unchanged in both sexes, whereas the differences between type 1 and type 2 diabetic subjects were significant only in men. After further adjustments for confounding factors, the average BMD values were still lower in type 1 diabetic subjects than in type 2 diabetic subjects although with lesser significance. Past low-energy fractures were more common in type 1 diabetic women than in type 2 diabetic women. CONCLUSIONS: The lower BMD in type 1 versus type 2 diabetic patients and control subjects probably results from more rapid bone loss after the onset of type 1 diabetes. This cannot be explained by insulin treatment, which was prescribed for both types of patients. Because the causes of low BMD in type 1 diabetes are unknown, these patients should be evaluated for the risk of osteoporosis and related fractures and offered appropriate preventive measures. PMID- 10388991 TI - Calcium antagonists and cardiovascular risk in diabetes. A review of the evidence. PMID- 10388992 TI - The European Association for the Study of Diabetes Annual Meeting, 1998. Treatment of type 2 diabetes and the pathogenesis of complications. PMID- 10388993 TI - Cultural adaptation of the diabetes quality-of-life measure for Chinese patients. PMID- 10388994 TI - Aminotransferase activity and acarbose treatment in patients with type 2 diabetes. PMID- 10388995 TI - High-dose vitamin C supplementation increases plasma glucose. PMID- 10388996 TI - Revised standards for the treatment of type 2 diabetes in Texas. PMID- 10388997 TI - Glycemic control and CAD risk: a heretical thought. PMID- 10388998 TI - Low-density lipoprotein particle size as an independent predictor of glycated low density lipoprotein level. PMID- 10388999 TI - Hypersensitivity to regular and intermediate, but not to crystallized, insulin as an aggravation factor for underlying bulimia nervosa in a patient with type 1 diabetes. PMID- 10389000 TI - Endothelial dysfunction is not reversed by simvastatin treatment in type 2 diabetic patients with hypercholesterolemia. PMID- 10389001 TI - Clinical features of elderly patients with type 2 diabetes. PMID- 10389002 TI - 2-year prospective audit of the effect of the introduction of insulin lispro in patients with specific clinical indications. PMID- 10389003 TI - Effect of fiber intake and meal pattern on gastrointestinal symptoms with acarbose. PMID- 10389004 TI - alpha-Tocopherol induces leptin expression in healthy individuals and in vitro. PMID- 10389005 TI - Shoulder MRI after surgical treatment of instability. AB - OBJECTIVE: To analyze magnetic resonance imaging (MRI) findings of the shoulder after an instability operation. MATERIALS AND METHODS: Physical examinations, radiographs and MRI of 10 patients after anterior glenoid bone block insertion for ventral instability were compared. MRI included T1-weighted spin-echo (TR = 600, TE = 20 ms) and T2*-weighted gradient-echo sequences (TE = 600, TE = 18, Flip = 30 degrees) in the axial, oblique-coronal and oblique-sagittal planes. RESULTS: No patient suffered from recurrent subluxation. We found fusion of the bone block with the anterior glenoid in seven cases, dislocation of the bone block without contact to the glenoid in one case, and no visible bone block in two cases. On MRI, the bone block showed either signal intensity equivalent to fatty bone marrow (n = 4) or was devoid of signal consistent with cortical bone or bone sclerosis (n = 4). In all patients, a low signal intensity mass, 2-4 cm in diameter, was visible next to the glenoid insertion site. CONCLUSION: Insertion of a bone block onto the anterior glenoid induces formation of scar tissue, increasing the stability of the shoulder joint. This scar is well visible on MRI and forms independently of the behavior of the bone block itself. MRI is ideally suited for evaluating postoperative shoulder joints after bone-grafting procedures. PMID- 10389006 TI - Magnetic resonance imaging or arthrography for shoulder problems: a randomised study. AB - OBJECTIVE: Diagnostic technologies are often assessed merely by their accuracy, rather than by their impact on diagnosis and patient management. To this end the authors have undertaken a study to assess the diagnostic and therapeutic impact of magnetic resonance imaging (MRI) and arthrography of the shoulder for patients referred from a rheumatology clinic. METHODS AND PATIENTS: Patients referred from a rheumatology clinic with symptoms warranting imaging of the shoulder were randomised to either MRI or arthrography. Data on the clinician's diagnostic confidence and management were recorded before and after imaging using questionnaires. Patients were followed-up at least 10 months after imaging to see how management plans evolved, and what proportion of patients required further imaging. RESULTS: Fifty three shoulders underwent imaging over a year and entered into the study; 29 randomised to MRI and 24 to arthrography. Both MRI and arthrography had a similar beneficial diagnostic impact in terms of clinical diagnoses (refuted and retained) and new diagnoses established. MRI and arthrography had a similar therapeutic impact, although MRI was associated with a significant shift towards surgical intervention. CONCLUSION: MRI and arthrography a have similar diagnostic and therapeutic impact. PMID- 10389007 TI - Acute swelling of the limbs: magnetic resonance pictorial review of fascial and muscle signal changes. AB - OBJECTIVE: This pictorial review analyzes the magnetic resonance (MR) fascial/muscular changes in 69 patients referred as emergencies with acute swelling of the limbs (ASL) from various causes. METHODS AND MATERIAL: A prospective MR imaging (MRI) study of 69 patients referred as emergencies for ASL was performed. Our population consisted of 45 patients with skin and soft-tissue infections (cellulitis and necrotizing fasciitis, and pyomyositis), six patients with soft-tissue inflammatory diseases (dermatomyositis, graft-versus-host disease), 11 patients with acute deep venous thrombosis, three patients with rhabdomyolysis, one patient with acute denervation and three other patients with rare diseases. Hematomas, tumorous or infectious bone involvement and soft-tissue tumors were excluded. All studies included spin echo T1-weighted images and spin echo T2-weighted images. Gadolinium-enhanced spin echo T1-weighted images were obtained when an abscess was suspected on T2-weighted images. Selective fat saturated T1- and T2-weighted sequences were also used. MRI analysis was performed to obtain a compartmentalized anatomical approach according to the location of signal abnormalities in subcutaneous fat, superficial and deep fascia and muscle. RESULTS: In all patients with ASL, MRI demonstrated soft-tissue abnormalities involving subcutaneous fat, superficial fascia, deep fascia, or muscle. Although MR findings were non-specific, MRI appears sensitive for detecting subtle fascial and muscle signal changes. CONCLUSIONS: In skin and soft tissue infections, MRI can be helpful for therapeutic management by determining the depth of soft-tissue involvement, particularly within fasciae and muscles, which is partly related to the severity of cellulitis with severe systemic manifestations. MRI can also aid the surgeon in diagnosing abscesses. In inflammatory diseases, MRI can determine the best site for biopsy and also monitor therapeutic response. PMID- 10389008 TI - Comparison of low field (0.2T) and high field (1.5T) MR imaging in the differentiation of torned from intact menisci. AB - PURPOSE: To evaluate the usefulness of a low field MRI system (0.2T; Esaote, Biomedica) for the evaluation of meniscal tears with regard to anatomic site, and to compare the results with findings from a high field unit (1.5T; Siemens, Erlangen). MATERIAL AND METHODS: MRI was performed in 25 patients in a low field (0.2T; Esaote, Biomedica), and a high field (1.5T; Siemens, Erlangen) MRI unit. The images were analyzed for the presence or absence of meniscal tears and the confidence of decision making. Results were further analyzed for the number of identical and unidentical findings on both imaging modalities. In seven patients, arthroscopy was performed and the findings compared with the results from MR imaging. Statistical analysis was performed by chi 2-test, Wilcoxon test and Friedman analysis. RESULTS: Qualitative evaluation of the level of confidence in decision making was significantly superior on high field strength images. When comparing the evaluations from both image modalities in 21 of 25 patients (84%), the diagnosis concerning the presence or absence of meniscal tears was identical. CONCLUSION: Although low field MR imaging might offer diagnostic potential concerning the presence or absence of meniscal tears, the level of confidence in decision making is significantly superior with high field strength imaging, probably reflecting the higher conspicuity of lesions from high field strength units. PMID- 10389009 TI - Bending stiffness of healing fractures can be calculated from quantitative computed tomography. AB - This paper revisits the relationship between bone densitometry and fracture healing by using quantitative computed tomography (QCT) to assess bone density. In recent time the correlation between bone mineral density (BMD) and mechanical stability of healing bone has been investigated with the purpose to predict mechanical properties of healing fractures--such as their loading capability- from data which can be collected non-invasive. One goal is to obtain a functional dependency between data obtained from QCT and the mechanical properties. In this study the computation of the mechanical stability of fractures from data obtained by QCT is proved to be reliable (r2 = 0.947 and P < 0.0001). As one result most dependencies between mechanical data and stiffness are not linear but quadratic (r2 > 0.72, P < 0.0005). The only linear dependencies are found between the second polar moment of inertia (Ip), calculated from geometric midpoint and center of mass (r2 = 0.688, P = 0.0052 and r2 = 0.677, P = 0.0010), and the average density (r2 = 0.836, P < 0.0001) versus bending stiffness. A functional dependency between bending stiffness and bone mineral content (BMC) of the fracture area in the fracture gap can be provided. The data presented in this work has been computed by a new algorithm developed by the author for detecting the fracture area of minimal density automatically in three-dimensional data obtained from QCT. PMID- 10389011 TI - Quiz case 6. Bronchiolitis obliterans organizing pneumonia (BOOP). PMID- 10389010 TI - Extramedullary paraspinal hematopoiesis in thalassemia: CT and MRI evaluation. AB - We present a comparative CT and MRI study of the paraspinal extramedullary hematopoiesis in 32 thalassemic patients. The patients were classified into four groups according to the MRI and CT imaging findings. Active recent extramedullary paraspinal hematopoietic masses show soft tissue behavior in both CT and MRI. Older inactive masses reveal iron deposition or fatty replacement. Combined imaging findings of paraspinal extramedullary hematopoiesis revealed the phase of its evolution and the correct diagnosis. PMID- 10389012 TI - Results of a prospective multicenter study for evaluation of the diagnostic quality of an open whole-body low-field MRI unit. A comparison with high-field MRI measured by the applicable gold standard. AB - OBJECTIVE: To evaluate the diagnostic quality of an open whole-body low-field MRI scanner compared to high-field scanners. MATERIALS AND METHODS: Over a period of 3 months, 401 patients with diseases of the kidney (n = 78), the shoulder (n = 122), the spine (n = 105) and the cerebrum (n = 96) were prospectively evaluated in four participating centers. They all underwent clinical evaluation, low-field and high-field MRI examination and surgical or follow-up confirmation of diagnosis. Clinical, histopathologic, high-field and low-field MRI diagnoses were recorded in standardized questionnaires that were centrally evaluated. Statistical evaluation comprised two parts: ROC analysis assessed accuracy of MRI and clinical diagnoses; furthermore rates of concordance of high- and low-field MRI diagnosis were calculated. RESULTS: We found no statistically relevant difference in high-field MRI diagnosis compared to low-field MRI diagnostic accuracy measured by clinical or surgical gold standard in three of the four regions examined; in cerebral examinations there was a small yet significant advantage for the high-field systems (P = 0.01). CONCLUSION: We conclude that the open low-field scanner we evaluated using clinical and surgical gold standard as reference is able to achieve comparable diagnostic accuracy compared to high field scanners at lower costs and greater patient comfort. Limitations due to field strength (signal-to-noise ratio, resolution, scan time) seem to be relevant only in a very small number of cases that warrant high-field examination. PMID- 10389013 TI - A Delphi study to establish national cost-effectiveness research priorities for positron emission tomography. AB - OBJECTIVE: This study aimed to determine the key cost-effectiveness research questions relating to positron emission tomography (PET) in the UK. METHODS: A systematic literature review was conducted to establish the existing knowledge base relating to the cost-effectiveness of PET in the various conditions for which it has been proposed. A three-round postal Delphi study of relevant individuals was used to determine the key cost-effectiveness research questions relating to PET in the UK. The content and structure of the Delphi study was informed by the results of the literature review. RESULTS: The most important cost-effectiveness research priorities for the National Health Service (NHS) relating to PET were in the clinical areas of lung cancer, breast cancer and the assessment of myocardial viability. Gamma camera PET using coincidence imaging was highlighted as a modality whose, clinical role needed to be determined urgently. CONCLUSION: Underlying the cost-effectiveness research priorities which were established is the need for evidence that the use of the various PET modalities as a diagnostic technique will alter patient management as compared to existing diagnostic strategies. The findings of the project provide a contemporary overview of the potential role for PET in the NHS and will be relevant to other countries. PMID- 10389014 TI - Evaluating scintillators used in radiation detectors of medical imaging systems by the effective fidelity index method. AB - OBJECTIVE: The performance of medical X-ray image receptors depends: (1) on the scintillator light emission efficiency; and (2) on the compatibility of the scintillator light spectrum with the spectral sensitivity of the light detector (film, photocathode, or photodiode), employed in conjunction with the scintillator. In this study, a scintillator performance measure, the effective fidelity index (EFI), is defined as function of both the scintillator light emission efficiency and spectral compatibility. MATERIALS AND METHOD: CsI:Na, Gd2O2S:Tb and La2O2S:Tb scintillators were employed in the form of phosphor screens prepared in our laboratory with various coating thicknesses. The screens were irradiated with X-rays employing tube voltages ranging between 50-120 kVp. RESULTS: The EFI performance of CsI:Na was found to increase with screen coating thickness and it was best when combined with the orthochromatic film or the ES/20 photocathode. Gd2O2S:Tb showed peak EFI performance at 70 mg/cm2 coating thickness and it was well combined with the light detectors considered. CONCLUSION: In accordance with our results, CsI:Na may be employed in radiography when adequately protected against humidity. Gd2O2S:Tb suitability for conventional imaging was verified and it was found that it may be useful in all types of digital imaging. La2O2S:Tb could also be used in digital detectors of imaging applications demanding medium X-ray tube voltages. PMID- 10389015 TI - Pacemaker and defibrillator Twiddler's syndrome. PMID- 10389016 TI - Erdheim-Chester disease. PMID- 10389018 TI - Oesophageal narrowing in chronic granulomatous disease. PMID- 10389017 TI - Imaging acute renal colic: 'mise au point'. PMID- 10389019 TI - The impact of antioxidants on chronic disease in ageing and in old age. AB - The antioxidant defense system is important in maintaining cellular homeostasis and preventing oxidative stress. Antioxidants are important dietary components contributing to a general slowing of ageing processes (e.g. cataract formation), but also in protecting particularly vulnerable sites from developing tissue injuries resulting in chronic diseases. This suggests that we should aim at certain minimum threshold concentrations of plasma antioxidants. In the near future, it is well possible that the amount of nutrients necessary to prevent or to reduce the risk for chronic diseases will be the more important issue than the amount it takes to prevent deficiencies. In order to achieve optimal nutritional levels, particularly elderly may benefit from the use of supplements during periods of stress or compromised nutrient intake. The highly interactive antioxidative system in the body strongly suggests that a well-balanced adequate intake of different antioxidants will be superior in its protective action to the supplementation with one single compound. It should be possible to resolve in the near future the question whether two days' RDA's are still adequate or whether the intake should be revised upwards. PMID- 10389020 TI - Physiology and pathophysiology of senescence. AB - Average life-span and maximum life-span are the two basic parameters by which the processes of ageing and senescence of individuals of a species are characterized. Although each individual of a species if affected by both parameters can only be studied in populations of individuals. The survival curve of a cohort of individuals reflects the different influences of constitutional and environmental factors on life expectancy. The intrinsic molecular and cellular mechanisms by which the physiological process of ageing and senescence is controlled and regulated are far from being understood although a large number of hypotheses have emerged over the decades. Stochastic and deterministic models of the ageing process have been developed but a theory unifying the large body of experimental, epidemiological and clinical findings is still lacking. In this contribution a brief review is presented on the different hypotheses aiming at explaining the physiology and pathophysiology of ageing and senescence. PMID- 10389021 TI - Vitamin status of elderly people in Germany. AB - In the last decade several attempts (Nationale Verzehrsstudie, NVS; Verbundstudie Ernahrungserhebung und Risikofaktoren-Analytik, VERA: Bethanien-Ernahrungsstudie, BEST) have been made to assess the nutritional status of the elderly in Germany. A careful evaluation of those data describing the vitamin status clearly indicate that healthy older people are not at higher risk for vitamin deficiency compared to younger adults. The results of the NVS showed that, except for folic acid, mean intake of all vitamins exceeded 80% of the current recommendations. Only 5% of blood vitamin concentrations analyzed in a subpopulation (VERA-Study) were founded to be below the physiological range. Only the incidence of low cobalamin values increased with age, presumably due to gastrointestinal problems (atrophic gastritis). In contrast, geriatric patients showed markedly lower vitamin blood concentrations compared to healthy subjects of the same age (BEST-Study). This might be explained by physical and mental deterioration, handicaps, chronic diseases and multiple chronic drug use. Underrepresentation of very old people, lack of reliable reference values for biomarkers and uncertainties in data collection may have contributed to misinterpretations. Representative studies are needed to objectively assess the nutritional status of the elderly population in Germany. PMID- 10389022 TI - The vitamin status and its adequacy in the elderly: an international overview. AB - Age-related changes in nutrition can affect the nutritional status of the elderly in a number of ways. Food intake is affected by socio-economic, physiological and pathological factors. The major physiological age-related change is the decrease in the energy requirement due to a reduction in lean body mass and a reduction in physical activity leading to a compensatory decrease in macro- and micronutrient intake of approximately 30% by the age of 80 years. Morbidity and some types of medication, smoking and alcohol consumption also affect the absorption and metabolism of vitamins. The plasma levels of fat-soluble vitamins and carotenoids tend to increase with age with the exception of vitamin D, while certain water soluble vitamin levels decrease, particularly vitamin B6 and vitamin B12. Many epidemiological studies have examined the vitamin intake and the plasma concentrations of large elderly populations in many regions of the world, but few have specifically determined the incidence of vitamin deficiencies. The criteria for defining deficiency varies between studies making it difficult to compare data from different studies. In the SENECA Study on European elderly evidence for biochemical vitamin deficiency was found in 47% for vitamin D, 23.3% for vitamin B6, 2.7% for vitamin B12 and 1.1% for vitamin E. PMID- 10389023 TI - Vitamin absorption in the elderly. PMID- 10389024 TI - Interactions between drugs and vitamins at advanced age. AB - Drug consumption increases at advanced age due to increased morbidity. At the same time the state of repletion is reduced for several vitamins. Physiological and kinetic alterations in the elderly are reviewed in order to analyse possible interrelations between these two phenomena. At high age the status of all vitamins is compromised by reduced food intake. Decreased active intestinal transport and an increased propensity for atrophic gastritis may reduce the absorption of vitamins A, B1, folate and B12. Decreased exposure to sunlight and reduced cutaneous synthesis impair the vitamin D status. Studies on the state of vitamin repletion in hospital patients indicate a specific response of vitamins A, B1, B6 and C to drug administration at advanced age. Reduced food intake in the elderly is further compromised by drugs that impair appetite and absorption. Anticonvulsives and other drugs that induce hepatic microsomal enzymes accelerate vitamin D metabolism and aggravate post-menopausal osteoporosis. Acid inhibiting agents increase achlorhydria and reduce vitamin B12 absorption. Renal clearance of acidic drugs such as acetylsalicylic acid and barbituric acid, which is impaired at high age, is further reduced by high doses of vitamin C. Vitamin B6 reduces the therapeutic effect of L-dopa. When recognised, the negative effects of drug-vitamin interactions can be compensated by adequate vitamin supplementation and by adaptation of drug dosing. PMID- 10389025 TI - The role of antioxidative vitamins in primary and secondary prevention of coronary heart disease. PMID- 10389026 TI - Lowering of homocysteine concentrations in elderly men and women. AB - B-vitamin supplementation has previously been shown to lower the concentration of plasma total homocysteine, a risk factor for cardiovascular disease. Little is known about the homocysteine-lowering effects of low-dose B-vitamins in elderly individuals, who are prone to higher homocysteine levels due to advanced age and a greater frequency of impaired vitamin status. We aimed to identify if and to what extent B-vitamins lower total homocysteine and its subfractions in elderly individuals. Men and women (> or = 60 years) received either B-vitamins (400 micrograms folic acid + 1.65 mg pyridoxine + 3 micrograms cyanocobalamin) or a placebo daily for 4 weeks. Subjects in the vitamin group showed a significant decrease in plasma total homocysteine during the first 2 weeks; thereafter, total homocysteine only slightly decreased further resulting in a geometric mean reduction of -16.3% (95% CI: -11.3% to -21.0%) over the entire treatment period. Free homocysteine decreased as well. However, the observed higher ratio of free/total homocysteine after 4 weeks of supplementation suggest a more pronounced reduction in protein-bound homocysteine. Low-dose B-vitamin supplementation is effective in lowering homocysteine in elderly individuals. Further studies are needed to be able to depict the effect of B-vitamin supplementation on different homocysteine sub-fractions in plasma. PMID- 10389027 TI - The role of vitamins in the prevention of osteoporosis--a brief status report. AB - This papers summarizes the main role vitamins are believed to play in the prevention of osteoporosis, a common disease which is anticipated to rapidly increase because of the aging of the population. Vitamin D, the classical vitamin related to bone health, improves bone strength mainly by increasing intestinal calcium absorption and reabsorption of calcium by the kidney. Several intervention studies demonstrated in humans that vitamin D can improve bone status as measured by bone density. Vitamin C is considered an essential cofactor of collagen formation. Epidemiological studies report a positive association between vitamin C intake and bone density. Intervention studies on the effect of vitamin C on bone status are missing. Vitamin B6 could function as a cofactor to build up cross-links. In humans, however, there is little evidence to support this. Vitamin K is required for the biological activity of several coagulation factors; the classical function of vitamin K. Recent research also points to a role of vitamin K in bone metabolism. Vitamin K mediates the <--carboxylation of glutamyl residues on several bone proteins, notably osteocalcin. Epidemiological studies and results from first intervention trials are consistently suggesting that vitamin K may improve bone health. PMID- 10389029 TI - Vitamin E in diabetes. Increased oxidative stress and its prevention as a strategy to prevent vascular complications? PMID- 10389028 TI - The potential preventive effects of vitamins for cataract and age-related macular degeneration. AB - Age-related cataract and age-related macular degeneration (AMD) are important public health problems. Approximately 50% of the 30 to 50 million cases of blindness worldwide result from unoperated cataract. In the US and other developed countries AMD is the leading cause of blindness, but age-related cataract remains the leading cause of visual disability. Age-related cataract and AMD represent an enormous economic burden. In the United States more than 1.3 million cataract extractions are performed annually at a cost of approximately $3.5 billion. Much of the experimental research on the etiology of cataract and AMD has focused on the role of nutritional antioxidants (vitamin C, vitamin E, and carotenoids). Evidence from epidemiologic studies support a role for nutritional antioxidants in delaying the onset of these age-related vision disorders. Although it is not yet possible to conclude that antioxidant nutrients have a role in prevention of cataract or AMD, a summary of the epidemiologic evidence suggests that it is prudent to consume diets high in vitamins C and E and carotenoids, particularly the xanthophylls, as insurance against the development of cataract and AMD. PMID- 10389030 TI - Vitamin E and other antioxidants in neuroprotection. AB - Several pathological conditions are believed to be causally related to the generation of reactive oxygen species and free radicals including various neurodegenerative disorders. In the histopathology of Alzheimer's disease (AD) many signs of oxidative reactions can be found building the basis of the oxidative stress hypothesis of AD. One major player in the generation of an overall oxidative microenvironment for the nerve cells is the amyloid beta protein (A beta) of the senile plaques in brain areas affected in AD. A beta can be neurotoxic and this toxicity is mediated by peroxides and by the peroxidation of membrane lipids leading to the lysis of the cell. Consequently, lipophilic free radical scavengers such as vitamin E and the recently discovered antioxidant activity of the female sex hormone estrogen protects neurons against the oxidative toxicity of A beta and other AD-related oxidative insults. In a first clinical trial using vitamin E in therapy, this antioxidant could slow down the course of the disease launching further clinical investigations. Although antioxidants act as non-specific protective chemical shields for neurons and do not target specific pathological events, they are highly effective and further investigations on their activity might lead to an even more effective application of antioxidants. Since the knowledge of the pathways of neuronal cell death that occur during oxidative challenges is increasing, it will be of central interest how antioxidants can interfere with signal transduction mechanisms and therefore also modify genetic programs. As long as specific interventions are not available the optimistic data concerning the neuroprotective activity of antioxidants in vitro and in vivo underline an important role for antioxidative acting compounds for the prevention and therapy of oxidative stress-related conditions including AD. PMID- 10389031 TI - Vitamin C, Helicobacter pylori infection and gastric carcinogenesis. PMID- 10389032 TI - Studies on vitamin B12 status in the elderly--prophylactic and therapeutic consequences. AB - Because of the large liver stores (about 5 mg), low turnover rate (0.143%) and small daily requirement (3 micrograms), vitamin B12 deficiency does not occur under normal circumstances. This is not the case in individuals with chronic inflammatory or trophic changes at vitamin B12 absorption sites. Without supplementation, vitamin B12 deficiency can be expected within 5 years of gastrectomy. Characteristic features of type A gastritis are hyposecretion and mucosal atrophy in the fundus and body of the stomach, with absent intrinsic factor. In the small intestine, active and/or passive absorption is impaired by extensive ileal resection, exocrine pancreatic insufficiency and chronic inflammatory disorders such as Crohn's disease. Definitive plasma concentrations cannot be quoted for vitamin B12 deficiency. Dietary habits, subjective symptoms, hematological laboratory results, function tests and gastrointestinal endoscopic and histological findings must all be taken into account in the diagnosis. Modern diagnostic parameters, such as methylmalonic acid and homocysteine serum assays, are useful for achieving early diagnosis and hence optimal treatment. With their assured availability, parenteral vitamin B12 preparations remain the treatment of choice. Results from vitamin B12 bioavailability studies in healthy subjects suggest that > 300 micrograms probably suffices as an oral maintenance dose after parenteral loading. Further well-documented cases are needed in order to establish whether these doses are adequate in malabsorption syndromes and gastrointestinal diseases. Various case reports indicate the value of prophylactic and therapeutic oral vitamin B12 administration, especially in disorders of homocysteine metabolism, a substance postulated as a further important risk factor for atherosclerosis. PMID- 10389034 TI - Factors affecting the results of the clock drawing test in elderly patients hospitalized for physical rehabilitation. AB - The Clock Drawing Test (CDT) is a recognized and accepted instrument for the early diagnosis of dementia in the elderly. In a prospective study we evaluated the association between the results of this test and a broad range of clinical, functional and sociodemographic variables. The study was conducted on elderly patients hospitalized for rehabilitation following stroke or hip fracture (HF) in the geriatric ward of a university hospital in southern Israel. The administration of the CDT and its scoring system were adapted from Sunderland et al. and Wolfe-Klein et al. The study was conducted on all 425 elderly patients who were hospitalized during the study period and who were capable of completing the test. Stepwise multiple regression was used to evaluate the association between the results of the CDT and the other variables. The mean CDT score (+/- SD) for the entire study population was 7.8 +/- 2.5 and 145 patients (34%) had scores of 6 or below. Of the 41 variables that were tested, significant associations with the CDT were found for the following four variables only: the Folstein minimental test (beta = 0.447, p < 0.0001), the cognition value from the admission FIM (beta = 0.252, p < 0.0001), years of education (beta = 0.183, p = 0.0001), and the patient's age (beta = -0.075, p = 0.037). The total variance of the CDT explained by these four variables (Adjusted R2) was 0.554. We conclude that in the study population there was a significant proportion of patients with low CDT scores. This score, in this population, is influenced in particular by two other measures of cognitive function and by the formal level of education, together with a weaker effect of age. PMID- 10389035 TI - Factors associated with depressive symptoms in non-demented community-dwelling elderly. AB - OBJECTIVE: We examined the risk for depressive symptoms associated with age, education, ethnicity, gender, marital status, apolipoprotein E genotype (APOE) and memory complaints among non-demented elderly (> or = 60 years). DESIGN: Cross sectional study of geriatric patients recruited from a free memory screening offered to the community. SAMPLE: This investigation included 506 community residing elderly subjects who were screened for cognitive impairment and classified as non-demented based on age and education-adjusted Folstein Mini Mental State Exam (MMSAdj) scores of 24 or greater. RESULTS: The prevalence of significant depressive symptoms (Hamilton Depression Rating Scale > or = 12) was 12.1% (N = 61). Increased risk for depression was associated with female gender (OR = 2.3; 95% CI = 1.1-4.8; p < 0.05), Cuban American ethnicity (OR = 4.9; 95% CI = 2.3-10.4; p < 0.0001) and memory complaints (OR = 1.3; 95% CI = 1.2-1.4; p < 0.0001). The APOE allele frequencies in the current sample were 0.07, 0.80 and 0.13 for the epsilon 2, epsilon 3 and epsilon 4 alleles, respectively. CONCLUSIONS: The results suggest that signs and symptoms of depression are common among non-demented elderly subjects in the community. In this study, mood disturbances were associated with Cuban American ethnicity, female gender and more memory complaints. Factors that were not confirmed by this study include age, education, marital status and APOE genotype. The observed APOE, epsilon 4 allele frequency of 0.13 supports the normal cognitive classification of the sample. PMID- 10389036 TI - The social consequences for families with Alzheimer's disease patients: potential impact of new drug treatment. AB - The social consequences of Alzheimer's disease are highlighted in this review with regard to impact on family situation, a changing treatment context caused by demographic changes, reorganization of long-term care, a financial crisis in the public health systems and the introduction of antidementia drugs. In the early phase of dementia there may be significant consequences for the patients and the family members which are largely unrecognized by the healthcare system. As the disease progresses, the impact on caregivers in terms of physical and emotional burden, financial and employment status may be enormous. The current care provision in Sweden, the UK and The Netherlands is described. Innovative care alternatives and strategies may improve the situation. The introduction of antidementia drugs such as the acetylcholine esterase inhibitors may also contribute to improved circumstances for patients and caregivers. There is still a great need for further research in this field. PMID- 10389037 TI - A descriptive survey of acute bed usage for dementia care in old age psychiatry. AB - OBJECTIVE: To examine the reasons why patients are admitted to acute dementia care assessment beds in one large district health authority (Leicestershire). DESIGN: A prospective questionnaire was completed by the RMO for every admission to dementia care assessment beds over a 6-month survey period. RESULTS: The most common reasons for admission were behavioural problems (most frequently aggression). Other common reasons included self-neglect, psychotic symptoms and comorbidity with functional psychiatric illness. Classification of patients by their social circumstances before admission indicated different patterns of presenting features, length of stay and place of discharge. CONCLUSIONS: The need for acute beds is demonstrated by the large number of emergencies, with a significant proportion admitted under the Mental Health Act, the type and complexity of reasons for admission and the preceding involvement of community mental health services. Multi-axial classifications including presenting problems and social circumstances, rather than the traditional method of looking purely at diagnosis, may offer some advantage in assessing outcomes. PMID- 10389038 TI - Measuring the decline of a population of people with early-onset dementia in Lothian, Scotland. AB - OBJECTIVE: To seek differences in the pattern of decline between groups of patients with early-onset dementia of varying aetiologies (clinically defined) on the basis of the investigation of their different clinical profiles. DESIGN: A cohort of 126 live patients with dementia diagnosed before the age of 65, of various aetiologies, were identified using the Lothian Psychiatric Case Register. Each person was seen for two assessments approximately 1 year apart. SETTING: Patients were either in long-term care (NHS or other) or at home, usually in the care of relatives. PATIENTS: Of 126 cases (53 male, 73 female, mean age at referral 58 years), 114 met the diagnostic criteria for DSM-III-R dementia: 60 Alzheimer's type dementia; 13 multi-infarct dementia; 14 alcohol-related dementia; with 25 in a mixed group of overlapping categories and two in 'other' dementia types. By the second assessment, 18 cases had died and two cases refused reassessment. MEASURES: The data collected included: demographic; behavioural and psychopathological; neurocognitive; neurological; and genetic. Change was assessed. RESULTS: The alcohol group was distinguished from the other groups by its generally milder profile of impairment and a pattern of change indicating some areas of improvement in the follow-up period. CONCLUSIONS: There are limitations in the instruments used to measure change in this group of patients; at present, measures of change in dementing illnesses cater predominantly for an elderly population with Alzheimer's disease. PMID- 10389039 TI - Ascertainment of a population of people with early-onset dementia in Lothian, Scotland. AB - OBJECTIVE: The aim of the case ascertainment for the study was to identify as near complete as possible a population of patients with presenile dementia who had been identified by hospital contact within a defined period of time, and who were currently alive, in the Lothian area. It did not aim to establish epidemiological figures. DESIGN: The Lothian Psychiatric Case Register was the main source of case identification. Case notes were inspected and cases identified for the study. SETTING: Patients were either in long-term care or at home. PATIENTS: As described in Paper I, Age range 30-65 years. MEASURES: Various diagnostic codings, including those of the ICD-9 were used to identify cases from the register. Feighner criteria were applied on case note inspection and DSM-III R criteria after individuals had been seen. RESULTS: A potential of 557 cases were identified from the case register, etc. Of these, 431 were excluded and 126 seen for the study. CONCLUSIONS: The criteria used for inclusion and exclusion of cases in this study are thus directed towards achieving the aims of this study, and the findings would have to be modified to reach an estimate of all cases of early-onset dementia in the Lothian area. PMID- 10389040 TI - Incidence of major depression in a very elderly population. AB - BACKGROUND: Depression is considered to be a major health problem in the elderly. Due to methodological problems, there are few studies on the incidence of depression in old age. The present study examines the prevalence of depression in a 3-year follow-up study of a non-depressed very elderly population, thus estimating the incidence. METHODS: 875 non-depressed persons with a mean age of 85 years were extensively examined by physicians twice with a 3-year interval. Depression diagnosis was made according to DSM-IV. All persons with a history of depression or a current depression were excluded in order to estimate the first incidence. RESULTS: 4.1% of the population was diagnosed as having a depression at the follow-up examination. The estimated first incidence was 1.4% per person year (0.8% in males and 1.5% in females). Characteristics at baseline correlated with the onset of depression were: having a dementia, insufficient social network and having more than two depressive symptoms. CONCLUSIONS: The incidence of depression was slightly lower in this very elderly population than for younger age groups, but followed the same female to male ratio. PMID- 10389041 TI - Detection of depression in elderly hospitalized patients in emergency wards in France using the CES-D and the mini-GDS: preliminary experiences. AB - OBJECTIVE: The purpose of this study was to evaluate the mini-GDS and the CES-D as instruments to detect depression in elderly hospitalized patients in emergency wards in France. METHODS: The CES-D was used on two cohorts of 60 non-cognitively impaired patients aged 70 or more. The mini-GDS was also used on the second of the two cohorts administered by a medical intern. These ratings were compared with a diagnosis of depressive disorder by ICD-10 criteria. RESULTS: The study population had a high (58%) prevalence of depression and a low level of active psychiatric referral. Mini-GDS and CES-D scores were well correlated (0.72, p < 0.001); the mini-GDS, with a cutoff score of 1, gave optimum sensitivity (88%) and specificity (63%). CONCLUSION: The use of the mini-GDS may aid the detection of depression in patients in emergency wards. PMID- 10389042 TI - Correlates of psychotic symptoms among elderly outpatients. AB - Psychotic symptoms presenting in late life can offer a diagnostic challenge to the clinician. In this study, 140 geriatric outpatients were prospectively examined for psychotic symptoms and assessed on a number of demographic and clinical variables. Cognition was assessed using the Mini-Mental State Exam. Psychiatric diagnoses were made by DSM-III-R criteria. Twenty-seven per cent (N = 38) had psychotic symptoms, delusions being the most common type. Patients with psychosis were significantly more likely to have a previous history of psychosis, to have a lower MMSE and to live in a nursing home. Four diagnoses accounted for 79.5% of all psychotic patients. In order of frequency, these were dementia, major depression, delirium and organic psychosis (organic hallucinosis, organic delusional disorder). Psychotic patients were significantly more likely to have a diagnosis of dementia, delirium or organic psychosis than non-psychotics, but depression was significantly more likely to occur in patients without psychosis. Although psychotic symptoms occur in a variety of illnesses, elderly patients with psychosis should be carefully evaluated for these disorders. PMID- 10389043 TI - Interpretative guidelines for neuropsychiatric measures with dichotomously scored items. AB - Neuropsychiatric measures consisting of dichotomously scored items are commonly used in clinical assessment. After summing these items, clinical guidelines frequently recommend cutoff scores to determine the presence or degree of a particular attribute, such as depression. However, blind application of such cutoffs neglects whether the total score is significantly different from chance. This confounding problem is illustrated using the Geriatric Depression Scale (GDS), and recommendations for interpreting the degree to which a GDS score significantly exceeds chance are presented. Specifically, GDS scores between 11 and 20, inclusive, were found not to differ significantly from chance (p > 0.05), assuming a random response pattern. The importance of supportive clinical evidence of depressive symptomatology is increased for scores in this range. These guidelines will be helpful in using such measures with patients who may vary with respect to response accuracy, and in assessing possible incomplete effort or random responding. PMID- 10389044 TI - Mobility and dementia: is physiotherapy treatment during respite care effective? AB - BACKGROUND: Mobility problems experienced by elderly people with a dementia are associated with falls, fractures and admission to long-term care. A hospital respite care admission is therefore often seen as an opportunity to provide physiotherapy treatment. AIM: To find whether elderly people with a dementia and a mobility problem show a greater improvement in mobility skills if given physiotherapy treatment than if given non-physical activities intervention during a hospital respite admission. METHOD: A controlled randomized multicentre trial with independent blinded assessment. The Southampton Mobility Assessment (mobility score) and Two Minute Walking Test (distance walked) were undertaken at the beginning and end of the study admission and beginning of the next respite admission. Following the first assessment, participants were randomized to either physiotherapy or activities. RESULTS: Eighty-one participants, from 12 clinical centres, with a mean age of 81.9 years and CAPE I/O score of 2. During the study admission there was a non-significant trend for a lower reduction in mobility score of the physiotherapy group (Mann-Whitney; p = 0.614) and a non-significant trend for greater decrease in distance walked in the activities group (t-test; p = 0.325). DISCUSSION: The results of this trial do not support the positive changes demonstrated elsewhere. However, changes in respite care during the early stages of this trial may have produced differences between the sample for this trial and that for the pilot study. This trial was therefore underpowered. CONCLUSION: This trial suggests that future research needs to change the focus from clinical settings to presentations. PMID- 10389045 TI - The phenomenology of delusional jealousy in late life. PMID- 10389046 TI - Depressive symptoms of Alzheimer caregivers. PMID- 10389047 TI - Alprazolam and hypotension. PMID- 10389048 TI - Vascular cognitive disorder. PMID- 10389049 TI - Growth and geographic distribution of selected health professions, 1971-1996. AB - This article examines the growth and geographic distribution of selected health professions in a 26-year period. The health professionals investigated were physicians, dentists, pharmacists, registered nurses, other health practitioners, dieticians and therapists, medical technologists and technicians, and health service workers. Allied health professions are represented by the last three of these groups. Samples of health professionals were extracted from the Current Population Survey from 1971 to 1996. The ratio technique and GIN1 index are used to describe the growth trend and geographic distribution of each health professional group over time. This historical overview reveals the following general trends in the 1990s: 1) growth of every selected health professional group has slowed down; 2) workforce disparities between the most and least abundantly supplied geographic areas have decreased; 3) selected health professions have become less evenly distributed among the population; and 4) the trends in pharmacy and dentistry call for immediate attention in workforce planning. The findings suggest that with the exception of dentistry, health personnel shortages are no longer an issue in such planning. Future health workforce policies should continue focusing on improving the distribution of workers among the general population. PMID- 10389050 TI - Advanced-practice sonography in obstetrics and gynecology: a pilot study investigating the efficacy of the ultrasound practitioner. AB - Advanced-practice (AP) ultrasonography by an AP sonographer or ultrasound practitioner is an emerging allied health profession. The efficacy of this professional in sonographic diagnosis has not been evaluated. This report describes a comparison of the efficacies of diagnosis using a traditional approach and diagnosis by the AP sonographer. Between 1991 and 1995, the authors performed a retrospective, case-controlled study. Patients were evaluated with either (1) a sonographer-based method (limited supervision with perinatal consultation as necessary) or 2) a sonologist-dominant method (direct supervision by a radiologist). Demographic information, ultrasonographic findings, delivery information, and data outcomes were compared. Multiple statistical methods were used and p < 0.05 was considered significant. Of 840 patients studied, 420 were evaluated with the sonographer-based method and 420 with a sonologist-dominant method. There was no significant difference in any potentially predisposing maternal risk factor (except maternal age and education), medical history, variable relevant risk factors associated with the current pregnancy, characteristics associated with delivery, or prenatal ultrasonographic variable. The evaluation of diagnostic efficacy for birth defects showed no difference. The sensitivity and specificity of diagnosis using the sonographer-based method were 100% and 94.7% for the detection of fetal abnormalities, respectively, and 100% and 91.4%, for the sonologist-dominant method. The diagnostic efficacy of the AP sonographer or ultrasound practitioner was found to be similar to that of the traditional sonographer-sonologist model. While, intuitively, there are significant advantages to multiple observers, the experienced, well-trained AP sonographer can function independently, with only discretionary consultation and assistance. PMID- 10389051 TI - Minority student persistence in clinical laboratory education programs. AB - The objective of this study was to characterize minority student persistence in clinical laboratory scientist (CLS) and clinical laboratory technician (CLT) education programs and relate persistence to student characteristics and involvement in academic and social dimensions of college programs. A prospective, longitudinal study was done using written survey and follow-up data collection. Participants were 2,426 CLS and CLT students in academic-based educational programs across the United States. The participants completed a forced-choice survey eliciting demographic information and determining their interactions with social and academic dimensions of college and the clinical laboratory education program. Program outcome groups, i.e., graduates, voluntary withdrawals, and academic dismissals, were compared on each of 14 variables. Of the 2,426 participants, 80.7% graduated, 10.8% voluntarily withdrew, and 8.5% were dismissed for academic reasons. The outcome groups differed significantly on seven of eight academic involvement measures and five of six measures of social involvement. Student persistence behaviors varied by ethnic group, with African Americans more likely to leave for academic reasons than others. Graduation rates varied among the five ethnic groups. Differences were found among ethnic groups on five of eight measures of academic involvement and on two of six measures of social involvement. The results suggest that ethnic groups may differ in their levels of involvement in aspects of social and academic programs, thus explaining their different persistence behaviors. Understanding why some students are unsuccessful may form the basis for effective retention efforts. PMID- 10389052 TI - Preceptor appraisals of rewards and student preparedness in the clinical setting. AB - The purposes of this study were to determine preceptors' expectations of students in the supervised practice setting; to identify rewarding and discouraging experiences in precepting; and to establish the perceived effect of the changing health care structure on precepting. A survey consisting of ten open-ended questions was mailed to 430 clinical instructors, representing five allied health programs. Results indicated that observing student growth was the most rewarding aspect of the preceptor role. Frustrating preceptor experiences included low student motivation and poor personal and professional behavior. Respondents expected students entering the clinical experience to have appropriate technical and communication skills. Increased pressures from health care restructuring are decreasing the internal rewards for preceptors. The findings of this study support both further preparation for students entering the clinical experience and a reward system for preceptors. PMID- 10389053 TI - Images within the allied health professions. AB - The purpose of this study was to identify the perceived images, understanding, and respect shared by employed members of six allied health professions and to determine the nature and extent of interactions these professions share as members of the hospital team. The results indicate a large variance in levels of understanding and respect among the professions. Most responded that they understood the other professions but the other professions did not understand them. The results also indicate that the professions interact on an average of no more than five hours per week, with their common interactive roles being peer relationships. PMID- 10389054 TI - Allied health applications of a computerized clinical log database system. AB - Preliminary research in the development and use of computerized clinical log records began in 1987 in an allied health college at a midwestern academic health center. This article reviews development and implementation of a computerized system for managing clinical log records to improve and enhance allied health educational programs in the radiation sciences. These clinical log databases are used for quantitative and qualitative analyses of student participation in clinical procedures, and educational planning for each student. Collecting and recording data from clinical log records serves as a valuable instructional tool for students, with both clinical and didactic applications. PMID- 10389056 TI - University-community partnerships for health: a model interdisciplinary service learning project. AB - This project is an example of a successful service-learning experiment at a major university. The program was successful in providing service-learning experiences for an interdisciplinary group of health-professions students, delivering essential health services to a community at risk, providing health-risk and demographic data, and offering opportunities for scholarly productivity for faculty. This was accomplished with a modest investment of internal start-up funding. Goals of the project were achieved, and the program and course were viewed as successful by students, faculty, and community partners. PMID- 10389055 TI - Systematic review of the effectiveness of interprofessional education: towards transatlantic collaboration. PMID- 10389057 TI - Seasonal affective disorder. PMID- 10389058 TI - Infection control--evidence into practice. AB - This paper arises out of two workshop sessions held at the fourth meeting of the Federation of Infection Societies, Manchester, 1997. The aims of the workshops were, first, to identify the factors which impede the process of translating research findings into infection control practice and second, to suggest how these barriers may be overcome. Key points from the workshops are presented within an idealized framework of creating, implementing and maintaining evidence based infection control practice. This lends structure to our exploration of the evidence underlying infection control guidance and the reasons why such guidance often does not result in appropriate action by healthcare workers. The strengths and weaknesses of each stage of the process are examined, using examples provided by participants at the workshop. PMID- 10389059 TI - Nurses and hospital infection control: knowledge, attitudes and behaviour of Italian operating theatre staff. AB - This study examined the disinfection and sterilization practices used by hospital operating theatres and evaluated the knowledge, attitude and behaviour of nursing staff with regard to infection control. Of the 216 nurses responding, knowledge concerning such practices was not consistent since 10% did not believe that items should be rinsed in water after contact with glutaraldehyde and more than 25% thought that 10 min contact time provided sterilization. Almost all were aware that improper practices increased the risk of nosocomial infections in patients. Nurses in orthopaedic surgery had a significantly lower level of knowledge compared with others. The great majority of nurses agreed that guidelines for disinfection and sterilization practice should be maintained and applied. With regard to the use of surgical instruments, the majority used steam or dry heat sterilizers for the appropriate time and temperature. Glutaraldehyde was used by 95% to sterilize endoscopes, but at different temperatures and times of exposure. Similar procedures were reported as used for laryngoscopes, though a higher percentage used heat sterilization. Only 38% routinely used all barrier techniques (gloves, masks, and protective eye-wear). Predictors for the routine use of all barrier techniques included attendance at continuing education courses on nosocomial infections, and nurses who were male and those involved in orthopaedic operations. Data support the need for finding and implementing interventions related to the prevention of hospital infection activities, in order to motivate nurses to use the correct procedures as a routine. PMID- 10389060 TI - Routine disinfection of patients' environmental surfaces. Myth or reality? AB - We have evaluated the need for daily disinfection of environmental surfaces not contaminated by biological fluids, in patient areas of a medical unit with two wings [North (N) and South (S)] at the University Hospitals of Geneva, Switzerland. Weekly bacteriological monitoring of surfaces was carried out at random (N = 1356 samples). In the S wing (control), we used detergent/disinfectant for daily cleaning of the floors and furniture. In the N wing we began by using a detergent for floors and furniture; after four weeks the results suggested changing to a rotation of detergent, dust attracting disposable dry mops and disinfectant. During this period the furniture was cleaned with an active oxygen-based compound. The average differences in contamination before and after cleaning floors were (mean reduction in bacterial counts and 95% confidence intervals; CI95): disposable mops: 92.7 cfu/24 cm2 (CI95; 74-112), active oxygen based compound 111.1 (90-133), and quaternary ammonium compound -0.6 (-27-26). Use of detergent alone was associated with a significant increase in bacterial colony counts: on average by 103.6 cfu (CI95 73-134). The quaternary ammonium compound was inadequate for disinfecting bathrooms and toilets but the active oxygen based compound was satisfactory. For furniture, there was a significant reduction in bacterial counts with both the methods using disinfectants. As the detergent was contaminated, by using it alone for cleaning, we were actually seeding surfaces with bacteria. A total of 1117 patients was studied and we observed no change in the incidence of nosocomial infections during the four months of the trial. In conclusion, uncontrolled routine disinfection of environmental surfaces does not necessarily make it safe for the patient and could seed the environment with potential pathogens. PMID- 10389061 TI - The effect of poor handling procedures on enteral feeding systems in Hong Kong. AB - Two enteral feeding systems commonly used in Hong Kong were evaluated for ease of bacterial entry during assembly and delivery of feeds. Wearing new, non-sterile disposable latex gloves during the assembly of the systems did not contaminate the feeds. The risk of contamination increased for systems assembled with bare hands. Systems assembled with hands experimentally contaminated with bacteria resulted in definite contamination of feeds. PMID- 10389062 TI - Are most ICU infections really nosocomial? A prospective observational cohort study in mechanically ventilated patients. AB - A prospective cohort study was undertaken with two end points: (i) to compare the 48 h time cut-off with the carrier state criterion for classifying infections, and (ii) to determine a time cut-off more in line with the carrier state concept. All patients admitted to the intensive care unit and expected to require mechanical ventilation for a period > or = 3 days were enrolled. Surveillance cultures of throat and rectum were obtained on admission and thereafter twice weekly to distinguish micro-organisms that were imported into the intensive care unit from those acquired during the stay in the unit. A total of 117 patients with median age of 61 years and median Simplified Acute Physiology Score II of 42, were included in the study. Of these patients, 48 (41%) developed a total of 74 infection episodes. Using the 48 h cut-off point, 80% of all infections were classified as ICU-acquired. According to the carrier state criterion, 44 infections (60%) were of primary endogenous development caused by micro-organisms imported into the intensive care unit. Seventeen secondary endogenous (23%) and 13 exogenous (17%) infections were caused by bacteria acquired in the unit. The carrier state classification allowed the transfer of 49% of infections from the ICU-acquired group into the import group. A time cut-off of nine days was found to identify ICU-acquired infections better than two days. These data suggest that monitoring of carriage of micro-organisms may be a more realistic approach to classify infections developing in the intensive care unit. PMID- 10389063 TI - Analysis of risk factors associated with nosocomial bacteraemias. AB - A prospective study of 2676 blood cultures was performed to identify the factors associated with clinically, significant nosocomial bacteraemia that occurred during a one year period in the Malaga University Clinical Hospital. Three hundred and fifty-five episodes of bacteraemia were considered clinically significant. The overall incidence of bacteraemia was 19.5/1000 admissions, of which 46% were hospital-acquired. A multivariate model showed that only six factors were significantly, and independently, responsible for nosocomial bacteraemias: intravascular catheterization (P < 0.0001, OR = 18.37), invasive procedures (P < 0.0001, OR = 10.38), malignancy (P = 0.035, OR = 3.11), indwelling devices (P = 0.005, OR = 3.05), stay in intensive care or surgical departments (P = 0.05, OR = 2.63) and length of hospital stay (P = 0.051, OR = 1.02). These results show that the factors which had most influence on the development of nosocomial bacteraemias were those factors associated with the treatment received by patients during their hospital stay. PMID- 10389064 TI - Efficacy of hand disinfectants against vancomycin-resistant enterococci in vitro. AB - Vancomycin-resistant enterococci (VRE) may be spread within a hospital via the contaminated hands of the healthcare worker. Effective hand disinfectants are necessary to break chains of transmission. We determined the bactericidal activity of 1-propanol, chlorhexidine digluconate (0.5 and 4%). Sterillium (45% 2 propanol, 30% 1-propanol and 0.2% mecetronium etilsulphate), Skinsept F (70% 2 propanol, 0.5% chlorhexidine digluconate and 0.45% hydrogen peroxide) and Hibisol (70% 2-propanol and 0.5% chlorhexidine gluconate) against 11 clonally distinct enterococcal isolates in a quantitative suspension test. Four isolates were vancomycin susceptible, four were vanA and the remainder vanB positive. Eight isolates were identified as Enterococcus faecium, two as Enterococcus faecalis and one as Enterococcus gallinarum. The investigator was blinded to the species and the genotype. Four parallel experiments were carried out for each isolate, each preparation, each dilution and each reaction time. 1-Propanol (60%), Sterillium, Skinsept F and Hibisol were all highly bactericidal after 15 and 30 s against VRE and vancomycin-susceptible enterococci (VSE) with reduction factors (RF) > 6.4, even in dilution of 50% (v/v). No significant difference was observed between vanA isolates, vanB isolates and VSE. Chlorhexidine digluconate (0.5% and 4%) was found to be less bactericidal after 30, 60 and 300 sec (RF < or = 2.5). The vanB genotype isolates were found to be significantly more susceptible to chlorhexidine (0.5%) than the vanA isolates (60 sec; one-way ANOVA model; P = 0.05). After 300 sec the vanB genotype isolates were found to be significantly more susceptible to chlorhexidine (0.5%) than the other two genotype isolates (P = 0.016). The vanA isolates were found to be significantly more susceptible to chlorhexidine (4%) than the vanB isolates (300 s; P = 0.024). E. faecium was found to be less susceptible to chlorhexidine than E. faecalis at all concentrations and reaction times, but significant differences between RF were only observed at 60 sec for both chlorhexidine concentrations (P < 0.05; t-test for independent samples). Propanol is much more effective against enterococci than chlorhexidine and combination of the two may be useful in providing an immediate and long lasting effect. PMID- 10389065 TI - Efficacy of Glucoprotamin containing disinfectants against different species of atypical mycobacteria. AB - A Glucoprotamin based disinfectant was tested against different strains of atypical mycobacteria in a suspension test. The disinfectant showed maximum detectable kill (> 4 log reduction factor) against all strains tested at 2500 ppm Glucoprotamin within 15 min, except for a glutaraldehyde resistant strain of Mycobacterium chelonae. For the latter, 2500 ppm for 60 min or 5000 ppm for 15 min, were necessary to achieve maximum kill. The glutaraldehyde resistant strain was also resistant to a mixture of different aldehydes. PMID- 10389066 TI - Detection of HGV in serum and peripheral blood mononuclear cells of maintenance haemodialysis patients. AB - The aim of the present study was to investigate the prevalence of hepatitis G virus (HGV) and also hepatitis C virus (HCV) infections in maintenance haemodialysis patients, and to identify extrahepatic sites as HGV reservoirs. HGV RNA was detected in the serum of 6/61 (10%) patients and in the peripheral blood mononuclear cells of 2/61 (3%) patients (one of whom was serum negative). These findings suggest that lymphoid cells constitute an extrahepatic HGV reservoir. HCV RNA was detected in 7/61 (11%) patients. Five of these patients (71%) were identified as carrying HCV genotype 1b. Co-infection with HCV and HGV was detected only in one patient. Haemodialysis patients are at risk for HGV infection, by nosocomial routes or via transfusions. HGV itself does not seem to be an important cause of hepatitis since all six HGV RNA positive patients not co infected by HCV or HBV showed normal ALT values. PMID- 10389067 TI - Impact of MRSA at a national cystic fibrosis centre. PMID- 10389068 TI - The myth of encrustation inhibiting materials. PMID- 10389069 TI - The quantity and duration of the antimicrobial efficacy of a chlorhexidine and silversulfadiazine impregnated catheter. PMID- 10389070 TI - Chlorhexidine dressing for reduction in microbial colonization of the skin with central venous catheters: a prospective randomized controlled trial. PMID- 10389072 TI - Prevalence of diarrhoea and vomiting on acute geriatric wards. PMID- 10389071 TI - Hospital environmental cleaning standards. PMID- 10389073 TI - Nosocomial aspergillosis during building work--a multidisciplinary approach. PMID- 10389074 TI - Posteroinferior glenoplasty can change glenoid shape and increase the mechanical stability of the shoulder. AB - The treatment of recurrent posterior glenohumeral instability remains an unsolved clinical problem. Although various types of capsulorraphy have been advocated, outcome studies indicate that it is difficult to achieve a balance between stability and mobility. Alterations of the bony glenoid for posterior instability have been proposed, but are not well understood from a mechanical perspective. This investigation had 2 purposes: (1) to determine in a cadaver model if posteroinferior glenoplasty can change the shape of the glenoid, and (2) to determine if altering the shape of the glenoid can increase the mechanical stability of the glenohumeral joint. We determined the effective glenoid shape in 7 normal cadaver glenoids by tracking the path of the center of the humeral head as it was translated across the glenoid face in 8 different directions. These determinations enabled us to calculate the maximum effective slope of the glenoid in each direction. We then determined the mechanical stability of the glenoids in each of the 8 directions by measuring the tangential force required to dislocate the shoulder under a 50-N compressive load. The ratio of the dislocating force to the compressive load was defined as the stability ratio. All measurements were repeated after a standardized posteroinferior glenoplasty was performed. Posteroinferior glenoplasty increased the posteroinferior glenoid depth from 3.8 +/- 0.6 mm to 7.0 +/- 1.8 mm and shifted the center of the humeral head an average of 2.2 mm anteriorly and 1.8 mm superiorly. These changes in dimension could be directly visualized as an immediate mechanical consequence of the glenoplasty procedure, particularly because of the insertion of the bone wedge. Glenoplasty increased the posteroinferior glenoid slope from 0.55 +/- 0.07 to 0.83 +/- 0.12 and increased the posteroinferior stability ratio from 0.47 +/- 0.10 to 0.81 +/- 0.17. This is a more than 70% increase in the tangential force that can be resisted before dislocation. The increase can be quantitatively understood as a direct mechanical consequence of the altered shape of the glenoid concavity. These numbers indicate that, in this cadaveric model, posteroinferior glenoplasty results in defined changes in the effective glenoid shape and in the mechanical stability of the glenohumeral joint. However, this study does not establish the role of this procedure in the clinical management of posterior glenohumeral instability. PMID- 10389075 TI - Arthroscopic management of the arthritic elbow: indications, technique, and results. AB - Twenty-four patients with painful restricted motion of the elbow joint because of an arthritic process were treated with an arthroscopic modification of the open Outerbridge-Kashiwagi procedure. Average preoperative flexion was to 90 degrees (range 60 degrees to 140 degrees), and average extension loss was -40 degrees (range -5 degrees to -60 degrees). The average total arc of motion was 50 degrees. The procedure consisted of arthroscopic debridement, partial resection of the coronoid and olecranon processes, and fenestration of the olecranon fossa. The radial head was excised arthroscopically in 18 of the 24 patients. All patients were reexamined 24 to 60 months after operation (mean 32 months). All patients had a significant decrease in pain as described by a visual analog scale (preoperative 8.2; postoperative 2.2). Average flexion was to 139 degrees (range 95 degrees to 145 degrees), and average extension loss was -8 degrees (range 0 degree to 15 degrees). The average arc of motion was 131 degrees, an improvement of 81 degrees. Arthroscopic ulnohumeral arthroplasty provides satisfactory results in terms of pain control and improved motion. The complication rate is comparable to those reported in series of open ulnohumeral arthroplasties. This procedure seems to be a valuable adjunct in the management of the arthritic elbow, serving as an intermediate step between nonoperative management and elbow replacement surgery. PMID- 10389076 TI - Primary anterior shoulder dislocation and rotator cuff tears. AB - In a prospective controlled study 167 patients with 167 primary traumatic anterior shoulder dislocations underwent early ultrasonograpic evaluation for rotator cuff tears. We found 53 (31.7%) full-thickness cuff tears in this group. Compared with a group of 93 healthy volunteers, we found with statistical significance more cuff tears in the patients aged < 60 years. Women ruptured the cuff more often than men. If the patient is not able to elevate the affected arm more than 90 degrees in the scapular plane 2 weeks after the dislocation, there should be a high suspicion of rotator cuff tear (76.7%). For early detection of relevant rotator cuff lesions, we recommend shoulder ultrasonographic examination and measurement of active elevation after each traumatic shoulder dislocation in the above mentioned age group. PMID- 10389077 TI - Arthroscopic relationship of the axillary nerve to the shoulder joint capsule: an anatomic study. AB - Twelve right shoulders in fresh cadavers were dissected to determine the relation of the axillary nerve to the shoulder capsule and glenoid. Needles transfixed the nerve to the capsule and into the shoulder joint. Arthroscopy was performed to determine the location of the needles on the glenoid clock. The needles were then removed and the position of the shoulder changed to determine the effect on the position of the axillary nerve. The axillary nerve was held to the shoulder capsule with loose areolar tissue in the zone between 5 and 7 o'clock and was close to the glenoid in the neutral position, in extension, and in internal rotation. With shoulder abduction, external rotation, and perpendicular traction, the capsule became taut and the axillary nerve moved away from the glenoid. Abduction, external rotation, and perpendicular traction increase the zone of safety during arthroscopic anteroinferior capsulotomy adjacent to the glenoid between the 5 and 7 o'clock positions. PMID- 10389078 TI - Arthroscopic subacromial decompression: results and factors affecting outcome. AB - Arthroscopic subacromial decompression was performed on 114 patients with rotator cuff impingement who had not responded to nonoperative measures. None of the patients had a full-thickness cuff tear. Patients with associated instability, symptomatic acromioclavicular joint disease, or ruptures of the long head of biceps were not included. Results were determined by questioning patients about their satisfaction with the outcome of surgery and by functional assessment of the shoulder with the parameters of pain, ability to perform daily activities, and range of motion according to the Constant scoring system. When reviewed at a mean interval of 19 months after surgery, 85 patients (75%) were satisfied with the outcome. Pain scores improved by an average of 8.6 points; "activities of daily living" scores improved by an average of 5.8 points; range-of-motion scores improved by an average of 3.6 points. The improvements in all 3 parameters scored were significant (P < .05). The following variables were statistically analyzed to assess their influence on final outcome: age, sex, occupation, duration of symptoms before surgery, dominance of the affected shoulder, outcome of the impingement test, state of the cuff, and experience of the surgeon performing the operation. The duration of symptoms before surgery was the most significant predictor of outcome. Symptoms of prolonged duration were associated with an unsatisfactory subjective results (P < .01) and with smaller improvements in the parameters of the Constant score (P < .001). Recovery after arthroscopic subacromial decompression and eventual outcome were related to the extent of cuff damage. Patients with partial thickness tears or fraying of the cuff had a delayed return to work (P < .001) and were found to have smaller increases in the pain and range-of-motion scores (P < .05). A satisfactory subjective result was most often associated with a positive impingement test (P < .05). Unsatisfactory outcomes were associated with a questionable diagnosis and lack of clear evidence of impingement at arthroscopy, inadequate decompression of the subacromial space, or the presence of calcium deposits in the rotator cuff. PMID- 10389079 TI - Elbow joint kinematics after excision of the radial head. AB - The contribution of the radial head to elbow joint kinematics was studied in 7 osteoligamentous elbow preparations. During unloaded flexion and extension, radial head excision induced a maximum varus displacement of 1.6 degrees with 20 degrees of joint flexion and a maximum external rotation of 3.2 degrees at 110 degrees of flexion. With application of a 0.75-Nm load, radial head excision induced a maximum laxity of 3.3 degrees at 20 degrees of flexion in forced varus and a maximum laxity of 8.9 degrees at 10 degrees of flexion in forced external rotation. No laxity was observed in forced valgus or internal rotation. The results were independent of the rotation of the forearm. This study indicates that the radial head acts as stabilizer to the elbow joint in forced varus and in forced external rotation. The results suggest that fractures of the radial head cannot be treated by simple excision without altering the basic kinematics of the elbow joint. PMID- 10389080 TI - Functional and magnetic resonance imaging evaluation after single-tendon rotator cuff reconstruction. AB - The aim of this study was to investigate tendon integrity after surgical repair of single-tendon rotator cuff lesions. In 31 patients, 31 single-tendon repairs were evaluated. Thirty-one patients were available for clinical assessment and magnetic resonance imaging (MRI) at follow-up. A standard series of MR images was obtained for each. The results of functional assessment were scored according to the system of Constant. According to MRI evaluation, 21 (68%) patients had an intact or thinned rotator cuff and 10 (32%) had recurrence of a full-thickness cuff defect at follow-up. Patients with an intact or thinned rotator cuff had a median Constant score of 75.5 points; patients with a full-thickness cuff defect had a median score of 62 points. There was no correlation between tendon integrity on postoperative MR images and functional outcome. Patients with intact or thinned cuffs did not have significantly better functional results than patients with retorn cuffs. Because of the presence of metal artifacts and the difficulty in distinguishing postoperative scar tissue from partial tears or thinning, MRI is of minor diagnostic value in assessing the shoulder after cuff repair. However, full-thickness tears are readily diagnosed after operation with MRI. PMID- 10389081 TI - Do functional deficits result from radial head resection? AB - The treatment of comminuted fractures of the radial head with primary or secondary head resection is controversial. To assess the outcome of patients after radial head resection, a retrospective study with clinical and radiologic follow-up including isokinetic testing was performed. Between 1981 and 1992, 151 patients underwent radial head resection for comminuted fractures. Fifty-nine patients were operated on during the first 2 weeks after injury (primary treatment), 47 patients were operated on between 3 weeks and 6 months after injury (early secondary), and 45 patients were operated on more than 6 months after injury (late secondary). Follow-up examinations of 108 patients were conducted at an average of 6 years after operation. In 64% of the patients only the radial head was fractured. In 26% of the patients the fracture was combined with a dislocation of the elbow. Results on the clinical and isokinetic tests were better for patients treated with primary resection than for patients treated with secondary resection. Of the patients treated with primary resection, 45% were subject to no restrictions in daily life and 64% had no limits at work. PMID- 10389082 TI - Glenohumeral osteoarthrosis after Putti-Platt repair. AB - The Putti-Platt capsulorraphy for recurrent anterior dislocation of the glenohumeral joint was performed in 139 shoulders between 1955 and 1985. Sixty six (46%) operated shoulders were studied with a mean follow-up period of 22 years (range 10 to 40 years). There were 52 shoulders of men and 14 shoulders of women; 45% of the shoulders were on the dominant side. Two patients underwent surgery on both shoulders. The average age of the patients was 49.3 years (range 33 to 74 years). Evaluation was based on patient history and the results of physical examination and radiography. The redislocation rate was low (only 3%), and 71% of the patients did not have pain, strength loss, stiffness, or instability in the operated shoulder. Radiographs were made of all shoulders, including the nonoperated shoulders. Osteoarthrotic changes of the glenohumeral joint were found in 40 (61%) shoulders. Arthrosis was mild in 23 (35%) shoulders, moderate in 13 (20%) shoulders, and severe in 4 (6%) shoulders. The rate of glenohumeral arthrosis is increased in patients who have undergone a Putti-Platt procedure and is positively correlated with the length of time since surgery. No correlation was found in this study between external rotation at 6 months after operation and the development of glenohumeral arthrosis. The number of dislocations before operation was correlated with the severity of arthrosis but not with its incidence. PMID- 10389083 TI - Surgical treatment of full-thickness rotator cuff tears in patients 40 years of age or younger. AB - Full-thickness tears of the rotator cuff are uncommon in the first 4 decades of life. A retrospective analysis was conducted of 19 consecutive patients who were 40 years of age or younger and had been treated surgically for a full-thickness tear of the rotator cuff. Sixteen patients (84%) recalled an acute injury that heralded the onset of symptoms. Five of the patients had sustained an initial glenohumeral dislocation. At an average follow-up of 5.7 years, all patients were evaluated with regard to pain, function, range of motion, strength, return-to work status, return-to-sport status, and overall postoperative satisfaction. After operation, 15 patients (79%) reported diminished pain relative to their preoperative level, and 12 (63%) of 19 were able to function with the extremity above shoulder level. Fourteen patients (74%) returned to full-time employment, and half returned to sporting activities. Thirteen patients (68%) reported subjective improvement with regard to daily functional activities after surgical intervention. The most favorable results were seen in those patients who had sustained an acute glenohumeral dislocation in conjunction with a full-thickness rotator cuff tear and underwent combined stabilization and repair. The outcome for patients who received worker's compensation was less favorable. PMID- 10389085 TI - Effect of steroid injections on the rotator cuff: an experimental study in rats. AB - The aim of this study was to evaluate the effects of repeated steroid injections into the subacromial space. Thirty rats were injected either 3 or 5 times with triamcinolone in a dosage equivalent to that given to human beings or 3 or 5 times with saline into the subacromial space. One rat received no injection. The supraspinatus and infraspinatus tendons were evaluated macroscopically and microscopically. Two different staining methods were used on each sample including hematoxylin eosin and Miller's elastin/van Gieson's solution. After 5 steroid injections, we found focal inflammation, necrosis, and fragmentation of collagen bundles in the tendon in 4 of 7 rats. The tendons of the controls showed a normal structure (P < .05). There were no pathologic changes among the rats that were injected with triamcinolone 3 times. These results show that repeated subacromial injections of triamcinolone may cause damage to the rotator cuff of the rat. This finding may indicate cautious use of subacromial steroid injections in human beings. PMID- 10389084 TI - Neurologic complications of surgery for anterior shoulder instability. AB - Two-hundred eighty-two patients underwent anterior reconstruction for recurrent glenohumeral instability between 1981 and 1991. Twenty-three patients (8.2%) had a neurologic deficit after surgery. Seven had sensory disturbances only; 16 had sensorimotor neuropathies (8 having multiple deficits designated as a diffuse plexopathy and 8 having a more defined deficit in 1 or 2 cords or peripheral nerves). Complete resolution occurred in 18 of the 23 patients. Four patients had a residual deficit (1 patient was lost to follow-up). Three had persistent sensory disturbances; 1 had permanent biceps weakness. None of these patients underwent surgical exploration. Older age (P = .045) and a Bankart lesion (P = .029) were associated with a neurologic complication. At an average follow-up of 8.7 years, 252 patients responded to a questionnaire regarding shoulder outcome, including 20 of the 23 patients with nerve injuries. The difference in the median Rowe score of those with and without nerve injury was not significant (P = .072). Neurologic injuries after anterior shoulder surgery presumably arise as a result of traction. The prognosis for neurologic recovery is generally good. Neurologic injury did not interfere with the outcome of the stabilization procedure. PMID- 10389086 TI - Cost-benefit comparison: holmium laser versus electrocautery in arthroscopic acromioplasty. AB - This prospective study was designed to measure the costs and benefits of using a laser rather than electrocautery for soft tissue resection during arthroscopic shoulder decompression. Forty-nine shoulders with refractory Neer stage II impingement (persistent fibrosis and tendinitis) were divided into 2 groups. The composition of the 2 groups was similar with regard to sex, worker's compensation status, dominant arm involvement, duration of symptoms, and length of conservative treatment. In one group, electrocautery was used to ablate the bursa and periosteum, release the coracoacromial ligament, and maintain hemostasis. In the other group, a laser was used in place of electrocautery. Patients had been evaluated preoperatively with 2 functional scoring systems. The patients were reexamined at 1 week and at 1, 2, 3, 6, and 12 months after surgery. There were no differences between the groups with regard to functional outcome or satisfaction. There was also no difference in terms of estimated blood loss or operative time. However, there was a statistically significant difference in total hospital charges between groups, with the laser group having a 23% higher hospital bill. On the basis of these results, it is concluded that there was no medical benefit to laser-assisted arthroscopic subacromial decompression but there was an increased monetary cost. PMID- 10389087 TI - Rotational dissociation of glenoid components in a total shoulder prosthesis: an indication that sagittal torque forces may be important in glenoid component design. PMID- 10389088 TI - Massive osteolysis of the shoulder (Gorham-Stout syndrome). PMID- 10389089 TI - Reconstruction of the medial collateral ligament with flexor carpi radialis tendon graft for instability after capitellocondylar total elbow arthroplasty. PMID- 10389090 TI - Functional magnetic resonance imaging neuroactivation studies in normal subjects and subjects with the narcoleptic syndrome. Actions of modafinil. AB - Functional magnetic resonance imaging (fMRI) can be used to detect regional brain responses to changes in sensory stimuli. We have used fMRI to determine the amount of visual and auditory cortical activation in 12 normal subjects and 12 subjects with the narcoleptic syndrome, using a multiplexed visual and auditory stimulation paradigm. In both normal and narcoleptic subjects, mean cortical activation levels during the presentation of periodic visual and auditory stimulation showed no appreciable differences with either age or sex. Normal subjects showed higher levels of visual activation at 10:00 hours than 15:00 hours, with a reverse pattern in narcoleptic subjects (P = 0.007). The group differences in spatial extent of cortical activation between control and narcoleptic subjects were small and statistically insignificant. The alerting action, and imaging response, to a single oral dose of the sleep-preventing drug modafinil 400 mg were then determined and compared with placebo in both the 12 normal (8 given modafinil, 4 placebo) and 12 narcoleptic subjects (8 modafinil, 4 placebo). Modafinil caused an increase in self-reported levels of alertness in 7 of 8 narcoleptic subjects, but there was no significant difference between mean pretreatment and post-treatment activation levels as determined by fMRI for either normal or narcoleptic syndrome subjects given modafinil. However, in the modafinil-treated group of 8 normal and 8 narcoleptic subjects, there was a clock time independent correlation between the initial level of activation as determined by the pretreatment scan and the post-treatment change in activation (visual, P = 0.002; and auditory, P = 0.001). No correlation was observed in placebo-treated subjects (P = 0.99 and 0.77, respectively). Although limited by the small number of subjects, and the lack of an objective measure of alertness, the findings of this study suggest that low cortical activation levels in both normal and narcoleptic subjects are increased following the administration of modafinil. Functional magnetic resonance imaging may be a valuable addition to established studies of attention. PMID- 10389091 TI - The effects of sleep inertia on decision-making performance. AB - Sleep inertia, the performance impairment that occurs immediately after awakening, has not been studied previously in relation to decision-making performance. Twelve subjects were monitored in the sleep laboratory for one night and twice awoken by a fire alarm (slow wave sleep, SWS and REM sleep). Decision making was measured over 10 3-min trials using the 'Fire Chief' computer task under conditions of baseline. SWS and REM arousal. The most important finding was that sleep inertia reduces decision-making performance for at least 30 min with the greatest impairments (in terms of both performance and subjective ratings) being found within 3 min after abrupt nocturnal awakening. Decision-making performance was as little as 51% of optimum (i.e. baseline) during these first few minutes. However, after 30 min. performance may still be as much as 20% below optimum. The initial effects of sleep inertia during the first 9 min are significantly greater after SWS arousal than after REM arousal, but this difference is not sustained. Decision-making performance after REM arousal showed more variability than after SWS arousal. Subjects reported being significantly sleepier and less clear-headed following both SWS and REM awakenings compared with baseline and this was sustained across the full 30 min. In order to generalize this finding to real-life situations, further research is required on the effects of continuous noise, emotional arousal and physical activity on the severity and duration of sleep inertia. PMID- 10389092 TI - Bright light treatment used for adaptation to night work and re-adaptation back to day life. A field study at an oil platform in the North Sea. AB - Night workers complain of sleepiness, reduced performance and disturbed sleep due to lack of adjustment of the circadian rhythm. In simulated night-work experiments scheduled exposure to bright light has been shown to reduce these complaints. Here we studied the effects of bright light treatment on the adaptation to 14 days of consecutive night work at an oil platform in the North Sea, and the subsequent readaptation to day life at home, using the Karolinska sleep/wake diary. Bright light treatment of 30 min per exposure was applied during the first 4 nights of the night-shift period and the first 4 days at home following the shift period. The bright light exposure was scheduled individually to phase delay the circadian rhythm. Bright light treatment modestly facilitated the subjective adaptation to night work, but the positive effect of bright light was especially pronounced during the re-adaptation back to day life following the return home. Sleepiness was reduced and the quality of day was rated better after exposure to bright light. The modest effect of bright light at the platform was, possibly, related to the finding that the workers seemed to adapt to night work within a few days even without bright light. These results suggest that short term bright light treatment may help the adaptation to an extended night-work period, and especially the subsequent re-adaptation to day life. PMID- 10389093 TI - Cardiac autonomic nervous system activity during presleep wakefulness and stage 2 NREM sleep. AB - Previous research has found that cardiac parasympathetic nervous system (PNS) activity increases and cardiac sympathetic nervous system (SNS) activity decreases during night-time sleep. This study aimed to examine in greater detail the time course of these changes in cardiac autonomic nervous system (ANS) activity. In the week prior to the experimental night, nine subjects maintained a constant sleep-wake schedule and experienced an adaptation night. Each subject's experimental night consisted of 2 h of presleep wakefulness, followed by a night of sleep, commencing at each subject's normal sleep onset time. One hundred and twenty beat blocks of presleep wakefulness and stable Stage 2 non-rapid eye movement (NREM) sleep across the night were selected. SNS activity was assessed using pre-ejection period, the amplitude of the T-wave in the ECG and the 0.1 Hz peak from the spectral analysis of the ECG. PNS activity was assessed using respiratory sinus arrhythmia (spectral analysis). Heart rate and respiratory rate were also measured. The results indicated a progressive decrease in SNS activity throughout sleep and a rise in PNS activity during the first half of the normal sleep period. The changes in PNS activity were similar, while the changes in SNS activity were altered, compared with a previous study in which stage of sleep was not controlled. This indicates a likely sleep stage influence on SNS activity, but not on cardiac PNS activity. These results are consistent with the concept of a primarily circadian, but not sleep, influence on PNS activity, and primarily a sleep, but not circadian, influence on SNS activity. PMID- 10389094 TI - Respiratory-related evoked potentials during the transition from alpha to theta EEG activity in stage 1 NREM sleep. AB - It has been argued previously that evoked potential components during Stage 1 sleep in response to both auditory and respiratory stimuli are intermediate between those of wakefulness and Stage 2 sleep. However, state fluctuations in the ECG between alpha and theta during Stage 1 sleep have been linked to changes in a number of respiratory functions including ventilation, upper airway resistance and chemical drive. It was therefore hypothesized that if respiratory related evoked potentials (RREP) were averaged separately for alpha and theta EEG periods during Stage 1 sleep, the alpha RREP would resemble wakefulness and the theta RREP would resemble Stage 2 sleep. RREPs were produced by 250 ms occlusions in 10 subjects. EEG was recorded from 29 scalp sites, referenced to linked ears, together with EOG and EMG. The N1 component was not specifically associated with alpha vs. theta activity, but appeared to be sensitive to any decrease in arousal level, suggesting that it was more related to attention than to changes in the EEG. The late N2 and P300 components were present during wake and Stage 1 alpha. However, in Stage 1 theta, different late components emerged (N300 and P450) that differed in latency, amplitude or topographical distribution from those seen in wakefulness. The P2 proved difficult to interpret, whereas the N550 did not appear until Stage 2 sleep, and as such, was not dependent on alpha/theta state. The results indicate that RREP components are differentially affected by the transition into sleep. PMID- 10389095 TI - Cardiac autonomic function during sleep in psychogenic and organic erectile dysfunction. AB - The present study investigated the sympathetic/parasympathetic balance during non rapid eye movement (NREM) and rapid eye movement (REM) sleep in patients with psychogenic and organic erectile dysfunction. The cardiac autonomic balance was assessed from the power of the low frequency (LF) and high frequency (HF) spectral components of heart-rate variability in 11 patients with psychogenic erectile dysfunction and 11 patients with organic erectile dysfunction as determined by monitoring sleep-related erections. Spectral analysis of heart-rate variability was calculated for at least four successive 4-min epochs of electrocardiogram recordings during NREM sleep and for all available 4-min epochs during REM sleep. Statistical analysis revealed that organic patients had a significantly higher LF/HF ratio (P < 0.01) during both stages of sleep, which resulted from a significantly lower power in the HF component (P < 0.004) and higher power in the LF component (P < 0.01) in these patients, in both REM and NREM sleep stages. These results demonstrate that patients complaining of daytime sexual dysfunction and found by sleep-related erection monitoring to suffer from organic erectile dysfunction, have altered cardiac autonomic balance during both stages of sleep. PMID- 10389096 TI - Obstructive sleep apnoea syndrome in hereditary gelsolin-related amyloidosis. AB - Gelsolin-related amyloidosis (AGel amyloidosis) is a rare autosomal dominant disorder, reported worldwide in kindreds carrying a G654A or G654T gelsolin gene mutation. The main clinical signs are cutis laxa, cranial and peripheral neuropathy, and corneal lattice dystrophy but heavy intermittent snoring also occurs. To evaluate whether sleep apnoea is present we performed nocturnal sleep recordings, cephalometric and spirometric analyses and multiple sleep latency tests (MSLT) in five snoring patients with a G654A gelsolin gene mutation. Four patients had obstructive sleep apnoea syndrome (OSAS) with redundant oropharyngeal and hypopharyngeal soft tissues, macroglossia and cranial neuromuscular dysfunction. The fifth patient had hypersomnia without obstructive sleep apnoea. Nasal continuous positive airway pressure (CPAP) was an effective treatment. This study presents the first evidence in favour of an association between AGel amyloidosis and OSAS, but further studies are needed to define the prevalence of OSAS and the pathogenetic roles of amyloid and variant gelsolin in its evolution. PMID- 10389097 TI - Sleep paralysis in the elderly. AB - Isolated sleep paralysis (SP) is a common sleep phenomenon that is highly colored by indigenous beliefs. In Hong Kong Chinese, the 'ghost oppression phenomenon' (GO) has been shown to be descriptively identical to SP. The prevailing concept is that the majority of cases with SP have their onset during adolescence, but the lack of any systematic study on an older population means that late-onset cases can not be excluded. In a study investigating the prevalence of mental disorders in Chinese elderly aged above 70 y in Hong Kong, we employed the revised GO questionnaire to study the prevalence of SP in this group of elderly as well. One hundred and fifty-eight subjects were finally analyzed for the study. Almost 18% (95% C.I. 11.77%, 23.68%) of the subjects reported experiences of GO. Their description of the features of GO showed striking similarity to those of SP. There was a clear bimodal distribution of onset of GO with peaks during adolescence and after age 60 y. At least one-third of the cases were late onset. In concordance with the rapid eye movement (REM)/wakefulness dissociation hypothesis of SP, those elderly with GO+ experiences also had more frequent nocturnal sleep disturbances. A family history was reported in 10% of subjects. PMID- 10389098 TI - Gender representation in sleep research. PMID- 10389099 TI - Psychophysiological insomnia, sleep mentation, and memory consolidation. PMID- 10389100 TI - Molecular approaches towards the isolation of sleep-related genes. AB - Behavioural genetics is one of the most enticing fields in modern biology. Owing to straightforward and semiautomated techniques that can be used to measure locomotor activity, circadian rhythmicity is perhaps the best studied behaviour in animals. Thus, during the past decade, five essential circadian clock genes have been isolated in Drosophila, and homologous counterparts for all of these genes have also been found in mammals. As the sleep-wake cycle is under the control of the circadian clock, these circadian master genes are expected to influence sleeping behaviour. However, different vigilance states are regulated by additional mechanisms that also have a genetic basis. In this article we discuss molecular approaches that may prove useful in the search for sleep related genes. PMID- 10389101 TI - EEG sleep patterns in twins. AB - The investigation of sleep in twins represents one of the major methods for measuring the genetic contributions to sleep in humans. This paper reviews two twin studies in which the sleep EEG was recorded during three consecutive nights in young monozygotic (MZ) and dizygotic (DZ) male twins. The analyses, based on average values of repeated sleep recordings, indicate that a significant proportion of variance in stages 2, 4, and delta sleep appears to be genetically determined. Genetic influences on rapid-eye-movement sleep were found inconclusive, but this conclusion is limited by the relatively small size of the sample studied. PMID- 10389102 TI - Genetic factors in human sleep disorders with special reference to Norrie disease, Prader-Willi syndrome and Moebius syndrome. AB - Sleep-wake problems are common in specific inborn errors of metabolism and structure of the central nervous system. Psychological factors, behavioural difficulties, metabolic disturbances, and widespread rather than focal damage to the nervous system are present in many of these diseases and all influence the sleep-wake cycle. However, a number of conditions cause relatively focal damage to the neuroanatomical substrate of sleeping and waking. These include fatal familial insomnia, with involvement of the prion protein gene on chromosome 20, Norrie disease, the Prader-Willi syndrome and the Moebius syndrome. The last three important conditions, although rare, are considered in detail in this review. They result in sensory deprivation, hypothalamic and mid-brain damage, and involve the X-chromosome, chromosome 15, and chromosome 13, respectively. These conditions cause a wide variety of sleep disturbance, including parasomnias, daytime sleepiness, and a condition like cataplexy. The place of the relevant gene products in normal sleep regulation needs further exploration. PMID- 10389103 TI - Fatal familial insomnia: clinical features and molecular genetics. AB - Fatal familial insomnia (FFI) is an autosomal dominant prion disease clinically characterized by inattention, sleep loss, dysautonomia, and motor signs and pathologically characterized by a preferential thalamic degeneration. FFI is linked to a missense mutation at codon 178 of the prion protein gene, PRNP, coupled with the presence of the codon methionine at position 129, the locus of a methionine-valine polymorphism. Homozygotes at codon 129, expressing methionine also in the nonmutated allele, have a shorter disease course (often less than 1 year), prominent sleep and autonomic disturbances at disease onset, and pathology restricted to the thalamus. Heterozygotes at codon 129, expressing valine in the nonmutated allele, have a longer disease course (often longer than 1 year), ataxia and dysarthria at disease onset, and lesions widespread to cerebral cortex. Both in the thalamus and in the cortex, the limbic structures are those most consistently and severely involved: the anterior ventral and mediodorsal thalamic nuclei, the cingulate gyrus, and the orbitofrontal cortex. FFI is thus a prion disease selectively damaging the thalamocortical limbic structures. Loss of sleep, sympathetic hyperactivity, and flattening of vegetative and hormonal circadian oscillations characterize FFI and result from a homeostatic imbalance caused by the interruption of the thalamocortical limbic circuits, the phylogenetically most advanced structures involved in the control of the sleep wake cycle and the body's homeostasis. The selective atrophy of the limbic thalamus that characterizes FFI might be due to the binding of FFI toxic PrP or PrPres to specific receptors on thalamolimbic neurons. PMID- 10389105 TI - Quantitative trait loci approach to the genetics of sleep in recombinant inbred mice. AB - Sleep is a complex trait controlled by many genes, the environment, and probably by gene-environment interactions. Among different approaches to the genetics of sleep, analysis of quantitative traits (QTL) has the advantage of being able to detect, along with major genes, minor and/or modifier genes influencing different quantitative aspects of sleep. We have used QTL analysis in two different sets of recombinant inbred (RI) strains and sought for confirmation of several localizations in eight histocompatibility congenic strains. Several QTLs were identified which influenced the amount of vigilance states. In a first RI series (seven strains) the only QTLs identified were those affecting paradoxical sleep (PS), whereas analysis in a second RI series (25 strains) revealed QTLs influencing PS, slow-wave sleep, and total sleep. Among these, a single QTL on chromosome 5 was associated with all vigilance states, suggesting the presence of a major gene influencing a basic aspect of sleep amount. Search for candidate genes around the identified QTLs indicated several immune related genes that have been implicated in sleep regulation. Transgenic animals carrying loss-of-function and/or gain-of-function mutations affecting these candidate genes should confirm these findings. PMID- 10389104 TI - Prion protein: a role in sleep regulation? AB - The prion protein (PrP) is a glycoprotein anchored to cell membranes and expressed in most cell types. Its structural features indicate possible relations to signal peptidases (Glockshuber et al. 1998). Since mutations in this protein lead to severe neurodegeneration and death in humans and animals, it is possible that the loss of its normal function contributes to the development of the pathology. Little is known about its normal function, but there are indications that it may play a role in circadian rhythm and sleep regulation in mice. We explored further whether PrP plays a role in sleep regulation by comparing sleep and the effects of 6 h sleep deprivation in PrP knockout mice and isogenic wild type mice of the 129/Ola strain. The mice did not differ in the amount and distribution of the vigilance states or in the power spectra. The most remarkable difference was the larger and long-lasting increase of slow-wave activity (mean EEG power density 0.75-4.0 Hz) in non-rapid-eye-movement (NREM) sleep during recovery from sleep deprivation in the null mice. The results confirm our previous findings in mice with a mixed background. This observation applies also to slow-wave activity in NREM sleep episodes following spontaneous waking bouts of different duration. Sleep fragmentation in both genotypes was larger than in mice with the mixed background. A new aspect was revealed by the spectral analysis of the EEG, where the null mice had a lower peak frequency within the theta band in REM sleep and waking, and not in NREM sleep. Behavioural observations concomitant with the EEG indicated that the EEG difference in waking may be attributed to the smaller amount of exploratory behaviour in the null mice. The difference between the genotypes in theta peak frequency was not an overall effect on the EEG, since it was absent in NREM sleep. PrP therefore may be affecting the theta-generating mechanisms in the hippocampus during waking and REM sleep. It remains unresolved whether PrP plays a role in sleep consolidation, nevertheless the data suggest that it is involved in sleep regulation. A passive avoidance test showed a difference between the genotypes. It is not probable that this was due to memory differences, since the genotypes reacted similarly in a delayed T-maze alternation procedure. The behavioural differences need to be pursued further. PMID- 10389106 TI - Differences in brain gene expression between sleep and waking as revealed by mRNA differential display and cDNA microarray technology. AB - The consequences of sleep and sleep deprivation at the molecular level are largely unexplored. Knowledge of such molecular events is essential to understand the restorative processes occurring during sleep as well as the cellular mechanisms of sleep regulation. Here we review the available data about changes in neural gene expression across different behavioural states using candidate gene approaches such as in situ hybridization and immunocytochemistry. We then describe new techniques for systematic screening of gene expression in the brain, such as subtractive hybridization, mRNA differential display, and cDNA microarray technology, outlining advantages and disadvantages of these methods. Finally, we summarize our initial results of a systematic screening of gene expression in the rat brain across behavioural states using mRNA differential display and cDNA microarray technology. The expression pattern of approximately 7000 genes was analysed in the cerebral cortex of rats after 3 h of spontaneous sleep, 3 h of spontaneous waking, or 3 h of sleep deprivation. While the majority of transcripts were expressed at the same level among these three conditions, 14 mRNAs were modulated by sleep and waking. Six transcripts, four more expressed in waking and two more expressed in sleep, corresponded to novel genes. The eight known transcripts were all expressed at higher levels in waking than in sleep and included transcription factors and mitochondrial genes. A possible role for these known transcripts in mediating neural plasticity during waking is discussed. PMID- 10389107 TI - Humoral regulation of physiological sleep: cytokines and GHRH. AB - Interleukin-1, tumour necrosis factor, and growth hormone releasing hormone form part of the humoral mechanisms regulating physiological sleep. Their injection enhances non-rapid-eye-movement sleep whereas their inhibition reduces spontaneous sleep and sleep rebound after sleep deprivation. Changes in their mRNA levels and changes in their protein levels in the brain are consistent within their proposed role in sleep regulation. Furthermore, results from transgenic and mutant animals also are suggestive of their role in sleep regulation. The sites responsible for the growth hormone releasing hormone somnogenic activity seem to reside in the anterior hypothalamus/basal forebrain. Somnogenic sites for interleukin-1 and tumour necrosis factor likely include the anterior hypothalamus, but also may extend beyond that area. These substances elicit non-rapid-eye-movement sleep via a biochemical cascade that includes other known sleep regulatory substances. PMID- 10389109 TI - Human insulin gene insertion in mice. Effects on the sleep-wake cycle? AB - Recently, insulin synthesis and the presence of an insulin receptor have been demonstrated in the brain. Intracerebroventricular infusion of insulin causes a selective increase in the amount of slow-wave sleep. In the present study, the sleep-wake cycle of transgenic mice, with or without habenular neuronal expression of the human insulin gene, was studied to investigate the possible role of brain insulin as a sleep modulator. Slow-wave sleep duration was increased in those mice expressing human insulin in the habenula. However, it is possible that this effect was not due to expression of the insulin transgene, but to the genetic background of one of the parental strains (CBA) used for insertion of the transgene. Users of transgenic mice should be aware of this possibility and be cautious in interpreting results when hybrid embryos are used as transgene recipients. PMID- 10389110 TI - The brain and the lymphatic system revisited. PMID- 10389108 TI - Prostaglandin D2 and sleep--a molecular genetic approach. AB - Prostaglandin (PG) D2 is the major prostanoid in the mammalian brain, and is the endogenous sleep-promoting substance in mice, rats, and monkeys, and probably in humans as well. When PGD synthase (PGDS), the enzyme responsible for the biosynthesis of PGD2 in the brain, was inhibited in vivo by its selective inhibitors, tetravalent selenium compounds, both slow-wave sleep and rapid-eye movement sleep were reduced almost completely but reversibly, indicating that PGDS is a key enzyme in sleep regulation. Experiments with transgenic mice also support this contention. In situ hybridization, immunoperoxidase staining, and direct enzyme assay of tissue samples revealed that PGDS is mainly, if not exclusively, localized in the arachnoid membrane and choroid plexus, from which it is secreted into the cerebrospinal fluid to become beta-trace protein. PGD2 exerts its somnogenic activity by binding with PGD2 receptors, exclusively localized at the ventro-rostral surface of the basal forebrain. CGS21680, an adenosine A2a agonist, mimicked the somnogenic activity of PGD2 when applied to the PGD2-sensitive zone. This effect was dose-dependently and selectively abolished by the prior i.p. application of the adenosine A2a antagonist KF17837. Furthermore, the somnogenic activity of PGD2 was also dose-dependently and selectively attenuated by KF17837, indicating the possibility that the sleep induction by PGD2 may be mediated by adenosine through A2a receptors under these conditions. When PGD2 was infused into the subarachnoid space below the rostral basal forebrain, concurrent with sleep induction, striking expression of Fos immunoreactivity was observed in the ventrolateral preoptic area. Fos expression in the ventrolateral preoptic area was positively correlated with the preceding amount of sleep and negatively correlated with Fos expression in the tuberomammillary nucleus. PGD2 also increased Fos IR in the basal leptomeninges and several regions implicated in autonomic regulation. These observations suggest that PGD2 may induce sleep via leptomeningeal PGD2 receptors with subsequent activation of the ventrolateral preoptic area neurons. PMID- 10389111 TI - The morphology of the lymphatics of the coronary arteries in the dog. AB - On the supposition that pericoronary lymphatics play an important role in the efflux of interstitial fluid from the blood vessel wall, we examined the morphology of pericoronary arterial lymphatics in the dog. After ligation of the principal epicardial drainage lymphatics, after ligation of the left anterior descending coronary artery, after induced pericoronary inflammation and after instillation of India Ink into the pericardial sac using light, dissecting, and electron microscopy. The findings were compared with non-operated (control) dogs. Lymphatic drainage of the coronary arteries is via adventitial lymphatics, which do not penetrate to the media and via periadventitial lymphatics consisting of a subepicardial lymphatic plexus overlying the coronary arteries. The smaller arterioles in the ventricular muscle have many more accompanying lymphatics than do epicardial coronary arteries. In the latter arteries, prelymphatic channels formed by collagen fibers in the media likely transport interstitial fluid to the adventitial and periadventitial lymphatics. Arterial contraction also likely plays a role in propulsion of coronary arterial interstitial fluid towards adventitial lymphatics. PMID- 10389112 TI - Bilateral groin adenolymphoceles: an unusual presentation of chylous reflux. AB - We report an unusual presentation of a young man with bilateral groin lymph nodal adenolymphoceles and right leg lymphedema as a manifestation of intestinal lymphangiectasia. Chylous reflux was supported by conventional and isotopic lymphography as well as by a total lipid test showing delayed triglyceride absorption 24 hours after ingestion of 60 gm of butter. After excision of groin masses in conjunction with dietary control (short-medium chain triglycerides), manual massage, pneumatic compression, and long-term wearing of a low stretch elastic garments he remains well. PMID- 10389113 TI - Age-related changes in the elastic fiber network of the human splenic capsule. AB - The structural arrangement of elastic fibers in the splenic capsule from 16 human cadavers ranging in age from 1 month to 76 years was studied by histologic sections stained with selective methods for elastin. In infants the elastic fibers of the splenic capsule were homogeneously intermingled with collagen fibers, an arrangement that stabilizes the capsule during spleen growth and enlargement. With aging, collagen fibers predominate in the outer capsular surface over elastic fibers with the latter more evident in the deep lamina of the splenic capsule. In elderly individuals, the elastic fibers shorten, fragment, and thicken. The progressive decrease in the amount of elastic fibers in the splenic capsule with aging may restrict splenic distention and contribute to involution of the spleen as one grows older. PMID- 10389114 TI - Structural studies of initial lymphatics adjacent to gastric and colonic malignant neoplasms. AB - Invasion and metastases are the main causes of death from cancer, and prognosis is best correlated with invasion of malignant cells into initial lymphatics and dissemination to regional lymph nodes. Using both light and transmission electron microscopy, we examined human gastric and colonic cancers and their relation to initial lymphatics. Invasion of malignant tumor cells into the initial lymphatics was characterized by interdigitating and overlapping endothelium giving way to open junctions as lymphatic endothelial cells were apparently dissolved and destroyed. Cytoplasmic vesicles, mitochondria, and rough endoplasmic reticulum were qualitatively increased as demonstrated by image analysis. PMID- 10389115 TI - Syncytial variant of nodular sclerosing Hodgkin lymphoma. AB - We report the case history of a 31-year old woman with a rare morphologic form of nodular sclerosing Hodgkin disease (NSHD) termed "syncytial variant." Its histologic features mimic metastatic carcinoma, thymoma, melanoma, non-Hodgkin lymphoma and germ-cell tumor. Antigens expressed on Reed-Sternberg cells, the hallmark of Hodgkin disease, and other neoplastic cells were screened to determine the correct diagnosis. This patient demonstrates the importance of using specific immunohistochemical techniques to clarify the diagnosis of NSHD of the "syncytial variant" subtype. PMID- 10389116 TI - Mechanisms involved in relaxation induced by exogenous nitric oxide in pig coronary arteries. AB - The aim of the present study was to analyze the mechanisms involved in the vasodilator responses elicited by nitric oxide (NO) in segments of porcine posterior descending coronary artery. Exogenous NO (0.1-30 microM) induced concentration-dependent relaxations in segments precontracted with a concentration of the thromboxane A2 mimetic, U-46619 (30-300 nM) that produced a contraction 70% (submaximal contraction) of that elicited by 75 mM K+ (maximal contraction). The relaxations were almost abolished by 6-anilinoquinoline-5,8 quinone (LY-83583, 10 microM), an inhibitor of guanylate cyclase, and with precontraction with 40 or 60 mM K+. Relaxations were reduced by 5 mM tetraethylammonium (TEA), a blocker of Ca(2+)-activated K+ channels (Kca channels) and unaltered by ouabain (500 microM), 4-aminopyridine (1 mM), glibenclamide (10 microM), apamin (1 microM) and charybdotoxin (0.3 microM), inhibitors of sodium pump, voltage-sensitive, ATP-sensitive, small-conductance Kca and large-conductance KcaK+ channels, respectively. These results suggest that the relaxation caused by exogenous NO is mediated by guanylate cyclase activation, with only a slight participation of a hyperpolarizing mechanism mediated by activation of Kca channels. PMID- 10389117 TI - L-NAME, a nitric oxide synthase inhibitor, modulates cholinergic antinociception. AB - Systemic administration of sumatriptan and buspirone (20 mg/kg: 5-HT1A agonists) produced antinociception against acetic acid-induced writhing. The antinociceptive effect was potentiated by cholinomimetic physostigmine (0.05 mg/kg i.p.) and blocked by the muscarinic antagonist atropine (5 mg/kg i.p.). Naloxone, an opiate antagonist, failed to reverse the sumatriptan- or buspirone induced antinociception, but pindolol (10 mg/kg), a nonselective 5-HT1A antagonist, blocked this response. Sumatriptan- or buspirone-induced antinociception was significantly potentiated by L-NAME (a nitric oxide [NO] synthase inhibitor) although L-NAME (20 mg/kg) given alone had no effect on the nociceptive threshold. Recent studies have suggested that the L-arginine/NO/cGMP pathway is involved in the modulation of pain perception. The present results suggest that NO may play a role in cholinergic antinociception-mediated 5-HT1A receptor stimulation and that NO exerts an inhibitory action on cholinergic analgesia. PMID- 10389118 TI - Intraperitoneal, but not subcutaneous, administration of the sulphated cholecystokinin octapeptide (CCK-8S) inhibits operant and nonoperant food intake in rats: implications for the CCK-satiety hypothesis. AB - The effects of administering CCK-8S by the subcutaneous (s.c.) and intraperitoneal (i.p.) routes were investigated on both operant and nonoperant food intake in rats that had been fasted for 22 h. Intraperitoneal administration of CCK-8S (5 micrograms/kg) significantly reduced both operant and nonoperant food intake. In contrast, CCK-8S (5-50 micrograms/kg) administered s.c. had no effects on food intake in both types of behavioral paradigms. The results show that the peripheral route of administration of CCK-8S is an important factor to be taken into consideration when investigating its effects on food intake. The results are discussed in terms of the hypothesis that CCK released from the small intestine during a meal acts in a paracrine fashion to produce satiety. PMID- 10389119 TI - Effects of anxiogenic drugs in rat forced swimming test. AB - The effect of antidepressants and anxiogenics in the forced swimming (Porsolt') test was investigated in rats. On the second day of an experiment, desipramine (10 mg/kg), pentylenetetrazole (20 mg/kg), picrotoxin (2.5 mg/kg), and clonidine (1.0 mg/kg) shortened while buspirone (1.0 mg/kg), yohimbine (2.5 mg/kg), DMCM (1.0 mg/kg), and Ro-15-4513 (1.0 mg/kg) prolonged the time of immobility or behavioral despair; fluoxetine (10 and 20 mg/kg), citalopram (10 mg/kg), and flumazenil (10 mg/kg) were ineffective. While clonidine, given in a subeffective dose (0.1 mg/kg), augmented the effect of desipramine (10 mg/kg), buspirone (1.0 mg/kg) had an opposite effect. The picrotoxin (2.5 mg/kg) challenge prominently shortened the time of immobility after desipramine (10 mg/kg) or citalopram (10 mg/kg) treatment. In conclusion, our results indicate that pharmacologically enhanced anxiety interacts with the effects of acute drug treatment in the forced swimming test. PMID- 10389120 TI - General pharmacological properties of the new proton pump inhibitor (+/-)-5 methoxy-2-[[(4-methoxy-3,5-dimethylpyrid-2-yl)methyl]sulf inyl]- 1H-imidazo[4,5 b]pyridine. AB - The general pharmacological profiles of a novel proton pump inhibitor, (+/-)-5 methoxy-2-[[(4-methoxy-3,5-dimethylpyrid-2-yl)methyl]sulfi nyl]- 1H-imidazo[4,5 b]pyridine, TU-199) on the central nervous system, cardiorespiratory system, autonomic nervous system, gastrointestinal system and renal functions were investigated. TU-199 had no effects on general signs and behavior in mice. TU-199 (300 mg/kg p.o.) decreased locomotor activity 3 h after administration in mice. TU-199 had no effect on pentobarbital-induced hypnosis, analgesic activity and electroshock-induced convulsion in mice, and on rectal temperature in rats. However, TU-199 (300 mg/kg p.o.) showed slight anticonvulsant activity on pentylenetetrazole-induced convulsion in mice. TU-199 had no effect on respiratory rate, blood pressure, heart rate, femoral blood flow and electrocardiogram in anesthetized dogs. TU-199 (10(4) M) caused the cumulative concentration-response curve obtained with acetylcholine in isolated guinea pig ileum to shift to the right. However, TU-199 showed no effect on contraction of isolated guinea pig ileum and had no effect on intestinal motility in mice, gastric emptying in rats, bile secretion in rats and carbachol-induced salivary secretion in mice. TU-199 had no effect on urinary volume and excretion of electrolytes in rats. These results suggest that TU-199 does not induce serious adverse effects on the central nervous system, cardiorespiratory system, autonomic nervous system, gastrointestinal system and renal functions with the exception of a decrease in spontaneous motor activity with high doses. PMID- 10389121 TI - Diminished facial expression despite the existence of pleasant emotional experience in schizophrenia. AB - In order to investigate the relationship between pleasant emotional experience and facial expression (i.e., laughter), mood before and after watching comic film clips, self-rated pleasant emotional experience for each film clip and electromyographic activities of facial muscles involved in laughter while watching film clips were measured for 25 patients with schizophrenia and 20 normal controls. Patients with schizophrenia who showed a significant correlation between self-rated emotional experience and major zygomatic activity were equivalent to normal controls in self-rated emotional experience and in mood after film clips; they had a significant increase in mood scores related with pleasure. Although these patients were thought to have sufficient pleasant emotional experience, they showed significantly low major zygomatic activity as compared to normal controls. It is suggested that these patients have a disturbance in the process of emotional expression rather than emotional experience. PMID- 10389122 TI - Redox dynamics of alpha-tocopherol in erythrocyte membranes of elderly patients with asymptomatic primary hyperlipidemia. AB - We investigated the redox dynamics of alpha-tocopherol in plasma and erythrocyte membranes in elderly patients with asymptomatic primary hyperlipidemia divided into three groups (hypercholesterolemia, hypertriglyceridemia and low high density lipoprotein cholesterol levels) and in healthy elderly subjects to assess the antioxidative status of alpha-tocopherol. alpha-Tocopherol and alpha tocopherolquinone were determined by high-performance liquid chromatography using a redox detection mode. In the erythrocyte membrane, there was no difference in the alpha-tocopherol concentration between hyperlipidemic and healthy subjects. The alpha-tocopherolquinone/alpha-tocopherol ratio in plasma and erythrocyte membrane, and the alpha-tocopherol in erythrocyte membrane/alpha-tocopherol in plasma ratio were significantly lower in elderly patients with hypercholesterolemia or hypertriglyceridemia. These findings suggest that the uptake ratio in erythrocyte membranes and the antioxidative activity of alpha tocopherol in both plasma and erythrocyte membranes are decreased in elderly hyperlipidemic patients. These decreases may promote membrane lipid peroxidation or accelerate atherosclerosis. PMID- 10389124 TI - The efficacy and safety of eperisone in patients with cervical spondylosis: results of a randomized, double-blind, placebo-controlled trial. AB - A randomized, double-blind, clinical trial was undertaken to assess the activity of eperisone hydrochloride (50 mg t.i.d.), a commonly used muscle relaxant, as a treatment for cervical spondylosis in 157 patients. The results showed a clear benefit of eperisone treatment with regard to pain in the nuchal region, back pain, pain in arms and shoulders, stiffness and other symptoms of cervical spondylosis, while the tolerability of the treatment was optimal. PMID- 10389123 TI - Tolerability and safety of 0.1% diclofenac plus 0.3% tobramycin fixed-dose ophthalmic solution: a randomized, comparative, controlled study in healthy volunteers. AB - The purpose of this double-blind, observer-masked, randomized, crossover trial was to compare the tolerability and safety of a fixed-dose ophthalmic solution of 0.3% tobramycin plus 0.1% diclofenac versus Tobrex (tobramycin sulfate ophth) and Voltaren (diclofenac sodium). Control treatments included a saline solution and a control solution of 0.3% tobramycin prepared by Alcon Cusi. Ten healthy volunteers received three consecutive instillations of 1 drop of a given ophthalmic solution at 08:00, 11:00 and 14:00 h to the same eye; after a washout period of 18 h, the next ophthalmic solution was tested according to a randomized sequence. Occurrence, intensity, and duration of ocular irritation and conjunctival hyperemia at baseline and after the three instillations were recorded. Slit lamp biomicroscopy examination, measurement of intraocular pressure (IOP) changes, visual acuity, and examination of the fundus of the eye were performed after each third instillation by an ophthalmologist. Side effect incidence and patient and investigator opinions were also recorded. Results showed that Voltaren instillation induced statistically significant ocular irritation (p = 0.0077); the remaining ophthalmic solutions tested caused no ocular irritation (Physiological Saline Braun, p = 0.9808; Tobrex, p = 0.8826; control 0.3% tobramycin solution, p = 0.8327; and 0.1% diclofenac plus 0.3% tobramycin, p = 0.5399). None of the ophthalmic solutions tested caused severe conjunctival hyperemia. Analysis of the sum of conjunctival parameters of both eyes for all ophthalmic solutions studied showed no statistically significant differences (p = 0.4688). Moderate superficial punctate keratitis was observed after instillation of Voltaren and of 0.1% diclofenac plus 0.3% tobramycin (1 subject each) that spontaneously resolved within 2 days. Slit lamp biomicroscopy, visual acuity and IOP values showed no statistically significant changes. No systemic side effects related to the study treatments were recorded. In conclusion, the ophthalmic solution containing 0.1% diclofenac plus 0.3% tobramycin was well tolerated under the study conditions. Its tolerability was equivalent to that of Braun physiological saline, Tobrex and a control 0.3% tobramycin solution and was better than that of Voltaren. PMID- 10389125 TI - Antagonism of the renin-angiotensin-aldosterone system and collagen metabolism in cardiac fibroblasts. AB - The renin-angiotensin-aldosterone system has emerged as a potential candidate for the accumulation of collagen in cardiac fibroblasts. The traditional renin angiotensin-aldosterone system can be considered a system in which circulating angiotensin II or aldosterone is delivered to target tissue or cells. However, an independent local renin-angiotensin system has also been described in cardiac cells and evidence has been accumulated for autocrine and/or paracrine pathways by which biological actions of angiotensin II can be mediated. These actions of angiotensin II are primarily mediated through angiotensin II receptors of the subtype I (AT1). When evaluating the effects of angiotensin II in situ, changes in circulating levels and local production both have to be taken into account. Functional angiotensin II receptors have been documented in cardiac fibroblasts although the presence of aldosterone receptors in cardiac fibroblasts is obscure, and the expression of mRNA for mineralocorticoid receptors in cardiac fibroblasts has been described. In vitro, angiotensin II increased cardiac fibroblast mediated collagen synthesis and mRNA levels of collagen type I, type III, pro alpha 1 (I) collagen, pro-alpha 1 (III) collagen and fibronectin, and inhibited matrix metalloproteinase I activity. The ability of angiotensin II to induce collagen synthesis and expression of collagen in cardiac fibroblasts may be mediated by an increase in transforming growth factor-beta 1 in an autocrine/paracrine fashion. The angiotensin II-stimulated secretion and expression of collagen was completely abolished by AT1 receptor antagonism, but not affected by AT2 receptor antagonism. The discordant findings that have been reported concerning the in vitro effect of aldosterone on collagen synthesis in cardiac fibroblasts can at least partly be attributed to differences in methodology such as the use of the total population or a sub-population of cardiac fibroblasts. In vivo, chronic infusion of angiotensin II or aldosterone increased the collagen volume fraction in the ventricles. Angiotensin-converting enzyme (ACE) inhibition and AT1 receptor antagonism, but not AT2 receptor antagonism, reduced collagen deposition in the myocardium in spontaneously hypertensive rats. The cardioprotective mechanism of action of ACE inhibitors can be attributed to local blockade of the formation of angiotensin II, to the degradation of bradykinin or to the release of nitric oxide and/or eicosanoids. Angiotensin-converting enzyme inhibitors also reduced collagen deposition in rat myocardium following myocardial infarction suggesting that collagen deposition may in part result from mechanisms other than through AT1 receptors. However, further research is necessary to unravel the various mechanisms involved in the action of angiotensin-converting enzyme inhibitors or of AT1 receptor antagonists on collagen deposition in the myocardium. PMID- 10389126 TI - Transdermal iontophoresis. Part II: Peptide and protein delivery. AB - The advent of sophisticated biotechnological processes has generated an interest in peptide and protein pharmaceuticals. However, the rapid developments in biotechnology are not matched by the developments in the delivery of these molecules. Presently, most of the peptides and proteins are delivered by parenteral route due to their poor oral bioavailability. The inherent discomforts associated with the parenteral therapy has prompted investigations into other nonparenteral routes and drug delivery techniques. Transdermal iontophoresis is one such technique showing good promise in the controlled and enhanced delivery of peptides and proteins. It offers noninvasive, continuous, pulsatile delivery as well as preprogramed complex dosing regimens. Miniature battery powered and wearable patches for peptides and proteins are expected to be on the market by the turn of this century. In continuation of our earlier review, this review explores the potential of transdermal iontophoresis for the delivery of peptides and proteins with the main focus on the suitability of the technique along with the factors to be considered in the design of this drug delivery system and its future prospects. PMID- 10389127 TI - Complications after intestinal transplantation: traditional and new. PMID- 10389128 TI - Beginnings and endings. PMID- 10389129 TI - Post-transplant lymphoproliferative disorders: a preventable complication of solid organ transplantation? PMID- 10389130 TI - Significance of Epstein-Barr virus status and post-transplant lymphoproliferative disease in pediatric thoracic transplantation. AB - Epstein-Barr virus (EBV)-associated post-transplant lymphoproliferative disease (PTLD) is a serious complication of organ and bone marrow transplantation. The importance of EBV matching between recipient and donor remains unclear. Between October 1987 and December 1997, 64 pediatric cardio-pulmonary transplants were performed at this center (58 hearts, two heart/lungs, four sequential single lungs). The EBV viral capsid antigen (VCA) immunoglobulin G (IgG) status of both donor and recipient was determined at the time of transplant. Of 56 patients from whom paired sera was available for analysis, 27 (50%) were recipient and donor EBV IgG positive, four (7%) were recipient EBV IgG positive and donor EBV IgG negative, and 12 (20%) were recipient EBV IgG negative and donor EBV IgG negative. The remaining 13 (23%) patients were EBV IgG negative but received organs from EBV IgG-positive donors. The EBV immunoglobulin M (IgM) status was determined from 6 weeks post-transplant in the 11 mismatches who survived for longer than 28 d. Seven became EBV IgM positive, two remained EBV IgM negative; the status of the remaining two remains unknown. The EBV IgM status was also determined retrospectively in patients who were EBV IgG negative pretransplant and received organs from EBV IgG-negative donors. Nine became EBV IgM positive; the rest remained negative. PTLD was diagnosed in two of 56 patients from whom paired sera was available; one was donor and recipient EBV IgG negative; the other was donor and recipient EBV IgG positive. No cases of PTLD were diagnosed in the remaining eight patients from whom paired sera was not available. Our experience suggests that PTLD does not occur with any greater frequency in the 'mismatch' group, and does not justify EBV matching in pediatric thoracic transplantation where there is a higher proportion of EBV-negative recipients than in adults. PMID- 10389131 TI - Living donor liver transplantation in critically ill children. AB - From December 1993, St Christopher's Hospital for Children, Philadelphia, PA, USA has provided living donors the opportunity to donate a portion of their liver to children who are critically ill. This report evaluates the results of living donor liver transplants (LDLT) in critically ill children. We retrospectively reviewed the first 22 LDLT at our institution and compared the patient and graft survival of the nine critically ill children with the 13 stable children. Twenty two LDLT have been performed at our institution between December 1993 and October 1997. Nine of 22 transplants [United Network for Organ Sharing (UNOS) Status I] were performed in children who were critically ill. Thirteen of the LDLT (UNOS Status II and III) were performed on stable children either in the hospital or admitted electively from home. The median weight and age at the time of transplant were 7 kg (range 4.6-54.5 kg) and 16 months (range 3 months-12 yr), respectively, and there was no statistical difference between the two groups. In critically ill children the 1-yr allograft and patient survival was 66% and 89%, respectively, exceeding the published results from UNOS for patients on life support (59.5% graft and 69.7% patient survival at 1 yr). One-yr allograft and patient survival in the stable children was 92.3% and 100%, respectively. All living donors are alive and well with normal liver function. In conclusion, our results show that LDLT is a viable approach for transplantation in critically ill children with liver failure and should be offered to potential donors. PMID- 10389132 TI - Duration of immunity to diphtheria and tetanus in young kidney transplant patients. AB - A considerable proportion of patients with renal transplant, evaluated many years after transplant, lack protective diphtheria antibody levels, despite primary immunization, but maintain immunity to tetanus. These patients respond to a diphtheria and tetanus booster but the duration of the response is uncertain. This study was undertaken to assess if protective antibodies evoked by primary immunization are lost quickly after transplantation, and whether the extent of the immune response to a booster influences the persistence of protective antibodies. We studied 15 patients (group 1) immediately after renal transplant and 35 patients with renal transplant for 6 +/- 4 yr who received a diphtheria and tetanus booster (group 2). Six patients (40%) of group 1 lost protective diphtheria antibodies a median time of 6.5 months after transplant. Thirty-three patients of group 2 responded to the booster with normal diphtheria antibody titers (> 1 IU/mL) in 22 cases and with low titers in 11. Four of the latter lacked immunity to diphtheria at 12 months follow-up. All patients with normal immunity maintained protective levels of diphtheria antibodies. The low responders had a creatinine clearance of 50 +/- 20 mL/min/1.73 m2. Tetanus immunity was maintained in almost all patients of both groups. In conclusion, renal transplant patients had an accelerated loss of diphtheria antibodies in the early post-transplant period. Response to a diphtheria booster identified a group at particular risk, namely the low responders, who may require frequent booster doses. This group had significantly poorer renal function than the normal responders. PMID- 10389133 TI - Long-term psychosocial adjustment following pediatric liver transplantation. AB - This study assessed long-term psychosocial adjustment to pediatric liver transplantation in 146 patients aged 4-25 yr, who had received a transplant 2-12 yr previously. Evaluations of psychosocial adjustment and related variables were based on the Harter Self-Perception Profiles for children, Child Behavior Checklist (CBCL), and children's academic level. Up to the age of 8 yr, transplant children as a group did not perceive themselves as less competent than healthy peers. Gender effects were characterized by older girls perceiving significantly less scholastic cognitive competence than their healthy peers. Adolescent and young adult boys had significantly lower global self-worth and lower perceived athletic competence than their healthy peers. In comparison to normative data of healthy children, CBCL parent-reported scores revealed significant deficits in competences for all age groups. Only for the older boys, however, did these deficits reported by the parents reach a pathological level. The majority of transplant children also had significantly higher problem scores, but they remained within the normal range, except for the older boys whose internalizing problems reached a borderline level. Our results suggest that liver transplantation does not substantially affect schooling. Regardless of statistically significant differences in psychosocial adjustment, the majority of the transplant children functioned at a normal level. For adolescent and young adult boys, however, the presence of problems and the lack of competences observed by parents and by the youngsters themselves reached borderline to pathological levels. Our findings stress the importance of psychological post transplant follow-up with increased attention of caregivers to child and parental concerns about their long-term psychosocial adjustment process. PMID- 10389134 TI - Diltiazem retards the metabolism of oral prednisone with effects on T-cell markers. AB - The area under the time-plasma concentration curve (AUC) was measured for prednisolone (the major active metabolite of prednisone) after ingestion of 15 mg of prednisone (phase 1) and again after 3 d of oral diltiazem (180 mg/d) followed by the same dose of oral prednisone (phase 2) in eight normal adult patients. Diltiazem increased the prednisolone AUC by 21% (range 3-38%), from 1297 +/- 157 ng/h/mL to 1560 +/- 169 ng/h/mL (p = 0.001). This effect was associated with a greater decrease from baseline in CD3+ lymphocyte number at 4 h after prednisone ingestion (596 +/- 175 vs. 516 +/- 140, p = 0.05), a larger percentage decrease of circulating CD3+ lymphocytes at 8 h (43 +/- 19% vs. 53 +/- 19%, p = 0.04), and a decrease in the number of CD3+ CD8+ T cells at 4 h post-prednisone ingestion (279 +/- 81 vs. 236 +/- 51, p = 0.04). Diltiazem retards prednisolone metabolism and when used chronically with prednisone could conceivably, in some patients, enhance its immunologic and other clinical effects. Potentiation of prednisone side-effects by diltiazem may be of special interest in pediatric patients, and possible diltiazem-prednisone interactions merit study in this population. PMID- 10389135 TI - Pathological evaluation of steroid withdrawal in pediatric renal transplant recipients. AB - Protocol biopsies were performed to detect and treat subclinical rejection in eight living related pediatric renal transplant recipients who had been withdrawn from steroids. Low-grade tubulitis (t1) and mononuclear cell interstitial inflammation (i1), typical of borderline rejection and low-grade interstitial fibrosis (ci1), and tubular atrophy (ct1), characteristic of chronic rejection, appeared frequently in protocol biopsies more than 3 yr after steroid withdrawal. In contrast, early-type glomerulitis (g), allograft glomerulopathy (cg), arteriolar hyaline thickening (ah), intimal arteritis (v), and fibrous intimal thickening (cv) were not observed in protocol biopsies after steroid withdrawal. Low-dose pulse therapy (methylprednisolone (MP) 250 or 500 mg/d for 3 d) was administered to five patients for borderline and/or acute grade 1a rejection, as determined by protocol biopsies in the absence of clinical rejection. No oral steroids were administered. Renal function in all patients remained satisfactory, without proteinuria, in follow-up periods ranging from 22 to 68 months after steroid withdrawal. Of the eight patients, three grew to almost normal height (> mean -2SD) and four exhibited catch-up growth. Thus, steroid withdrawal can be safe and improves growth in pediatric renal transplant recipients if undertaken with careful clinical follow-up, including protocol biopsies. PMID- 10389136 TI - Experience of parenteral nutrition for nutritional rescue in children with severe liver disease following failure of enteral nutrition. AB - Nutritional support is often necessary in chronic liver disease in childhood, and when enteral nutrition is insufficient, parenteral nutrition (PN) can be envisaged as a last resort. Pediatric experience is still limited in this indication. Seven children with severe liver disease received PN for a mean duration of 105 d, with additional enteral nutrition. Clinical tolerance was assessed and anthropometric and biological data were compared at the beginning and at the end of the study by the paired non-parametric test of Wilcoxon. Weight change, expressed as weight-for-age or weight-for-height Z-scores, increased. Conjugated bilirubin increased significantly. This retrospective study suggests that PN is a well-tolerated method for maintaining nutritional status in pediatric chronic liver disease when enteral nutrition has failed. PMID- 10389137 TI - Plasmapheresis, intravenous cytomegalovirus-specific immunoglobulin and reversal of antibody-mediated rejection in a pediatric renal transplant recipient: a case report. AB - This is a pediatric case report illustrating the development of antibody (Ab) mediated rejection in a patient with low levels of pretransplant anti-human leucocyte antigen (HLA) panel reactive antibodies (PRA). The clinical course of this patient suggests that aggressive use of a combination of plasmapheresis, monoclonal anti-T-lymphocyte antibody therapy, and intravenous immunoglobulin (IVIG) therapy can reverse Ab-mediated rejection in previously allosensitized pediatric transplant recipients. PMID- 10389138 TI - Heart transplantation on the first day of life from an anencephalic donor. AB - Heart transplantation on the first day of life, and graft harvesting from anencephalic donors, have been very rare events in the history of transplantation. At Bambino Gesu Hospital (Rome), heart transplantation was performed on a newborn 9 h after birth, using a graft harvested from an anencephalic donor. This graft achieved a good cardiocirculatory function, but the recipient died of necrotizing enterocolitis (NEC) on post-operative day (POD) 10. Despite failure, this case and other reports support the concept that hearts from anencephalic donors can work normally, and indicate that heart transplantation on the first day of life may have a favorable outcome if postoperative maintenance of multi-organ balance and function is successful. PMID- 10389139 TI - The 1997 Annual Renal Transplantation in Children Report of the North American Pediatric Renal Transplant Cooperative Study (NAPRTCS). AB - This report of the North American Pediatric Renal Transplant Cooperative Study (NAPRTCS), covering the years 1987-96 (January 15, 1997), analyzed data on 4898 patients who received 5362 transplants. Over the 10 yr of the study, 21.3% of the patients were under the age of 6 yr. Of the disorders that lead to end-stage renal disease (ESRD), structural and developmental anomalies of aplasia, dysplasia, and obstructive uropathy accounted for over 1500 patients. Despite the potential therapies for focal segmental glomerulosclerosis (FSGS), there has been little change in its incidence and this lesion continues to be the most common cause of renal failure and transplantation among acquired diseases. Eighty-nine per cent of patients with a functioning graft continue to receive cyclosporin A (CsA) 5 yr post-transplantation, and 84% of patients continue to receive azathioprine (AZA), whereas 26% of patients receive alternate-day steroid therapy at 4 yr post-transplant. Thirty-seven per cent of the living donor (LD) recipients and 47% of cadaver donor (CD) recipients were treated with the polyclonal T-cell antibody ATG/ALG for induction, and the monoclonal T-cell antibody, OKT3, was utilized for induction in 12% of LD patients and in 19% of CD patients. Twenty-five per cent of the 5362 transplants (1333) have failed over the 10-yr period. Graft survival is 90% at 1 yr and 74% at 6 yr for LD kidneys, and is 80% at 1 yr and 58% at 6 yr for CD patients. The most common cause for graft failure (30%) is chronic rejection. For recipients of LD grafts, relative risk (RR) factors for graft survival are African-American race (RR = 2.1, p < 0.001) and greater than five prior transfusions (RR = 1.6, p < 0.001). Prophylactic anti-T-cell antibody reduces the risk of graft failure (RR = 0.76, p = 0.009). For recipients of cadaver kidneys, risk factors are: recipient age < 2 yr (RR = 2, p < 0.001), donor age < 6 yr (RR = 1.3, p = 0.005), and absence of induction T-cell antibody (RR = 1.31, p < 0.001). PMID- 10389140 TI - Melatonin: from biochemistry to therapeutic applications. AB - Melatonin (N-acetyl-5-methoxytryptamine) is an evolutionary highly conserved molecule that plays an important role in conveying the clock and calendar information to all living organisms, including man. The hormone is synthesized mainly by the pineal gland, and, to a lesser extent, by extrapineal tissues--such as the retina, Harderian gland, and gastrointestinal tract. The melatonin generating system is characterized by three basic features: (1) photosensitivity, (2) diurnal (or circadian) rhythmicity (with highest levels of the hormone production occurring at night in darkness), and (3) age-related decrease in its activity. This review surveys data on the regulation of rhythmic melatonin biosynthesis by an array of factors, such as circadian pacemaker, light, neurotransmitters, second and third messenger molecules. Recent developments in the field of melatonin receptors are also presented. Finally, physiological and therapeutic properties of melatonin, with a special emphasis given to possible applications of this compound in the treatment of circadian rhythm sleep disorders, are discussed. PMID- 10389141 TI - Role of MAO A and B in neurotransmitter metabolism and behavior. AB - MAO (monoamine oxidase) A and B are key isoenzymes that degrade biogenic and dietary amines. MAO A preferentially oxidizes serotonin (5-hydroxytryptamine, 5 HT) and norepinephrine (NE), whereas MAO B preferentially oxidizes phenylethylamine (PEA). Both forms can oxidize dopamine (DA). However, the substrate specificity overlap and the in vivo function of these two isoenzymes is not clear. Recently, we have shown that MAO A and B knock-out (KO) mice exhibit distinct differences in neurotransmitter metabolism and behavior. MAO A KO mice have elevated brain levels of 5-HT, NE and DA and manifest aggressive behavior similar to men with a deletion of MAO A. In contrast, MAO B KO mice do not exhibit aggression and only levels of PEA are increased. Both MAO A and B KO mice show increased reactivity to stress. Taken together, these results suggest that MAO A and B have distinctly different roles in monoamine metabolism. Further, these mice are valuable models for investigating the role of monoamines in psychoses and neurodegenerative and stress-related disorders. PMID- 10389142 TI - Enzyme-catalyzed bioactivation of cyclic tertiary amines to form potential neurotoxins. AB - The pyridinium metabolites formed in the MAO-B catalyzed oxidation of 1-methyl-4 substituted-1,2,3,6-tetrahydropyridinyl derivatives, such as the parkinsonian inducing agent 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), cause the selective degeneration of nigrostriatal neurons, presumably by inhibition of mitochondrial respiration and depletion of ATP stores. The possibility that other partially oxidized piperidinyl derivatives also may be biotransformed to toxic pyridinium metabolites has led us to examine the metabolic fate of the neuroleptic agent haloperidol (HP) and its tetrahydropyridinyl dehydration product 4-(4-chlorophenyl)-1[4-(4-fluorophenyl)-4-oxobutyl]- 1,2,3,6 tetrahydropyridine (HPTP). In vitro metabolic studies employing tissue preparations isolated from rodents, baboons and humans have documented that cytochrome P4503A enzymes catalyze the biotransformation of both HP and HPTP to yield the corresponding pyridinium metabolite HPP+. An analogous biotransformation profile has been observed with "reduced haloperidol" (RHP), an abundant, circulating metabolite of HP formed by the stereospecific reduction of the benzoyl carbonyl group of HP. In vivo studies also have documented these pathways in humans, baboons and rodents. Although both HPP+ and RHPP+ are found in the urine and plasma of HP treated patients and HP or HPTP treated baboons, attempts to identify an MPTP-type lesion in baboons following long-term treatment with HPTP have failed. On the other hand, evidence for a lesion of the nucleus basalis of Meynert has been obtained. Additionally, the urinary excretion of abnormal organic acids and acylcarnitine conjugates suggests that HP and/or metabolites derived from HP interfere with energy production pathways. PMID- 10389143 TI - Interactive modulation by dopamine and serotonin neurons of receptor sensitivity of the alternate neurochemical system. AB - Ontogenetic dopaminergic denervation of rat forebrain is associated with latent supersensitization of dopamine (DA) receptors that is unmasked only by a priming process entailing repeated DA agonist treatments. Similar denervation supersensitivity holds for serotonin (5-HT) and most other neurochemical systems. Because DA and 5-HT neurons compete for target sites in the brain and mimic or replicate actions of the others, we investigated the modulatory influence of DA neurons on 5-HT receptor sensitivity; and role of 5-HT neurons in modulating DA receptor sensitivity. In these studies rats were lesioned with the DA neurotoxin 6-hydroxydopamine (6-OHDA, i.c.v.; desipramine pretreatment) or 5-HT neurotoxin 5,7-dihydroxytryptamine (5,7-DHT, i.c.v.; desipramine pretreatment) either at 3 days after birth or in adulthood. Responses to DA and 5-HT agonists were determined in several behavioral paradigms in adulthood. In assessing oral responses to agonists, we found that the D1 agonist (+/-)-1-phenyl-2,3,4,5 tetrahydro-1H-3-benzazepine-7,8-diol (SKF 38393) profoundly induced activity if rats were lesioned neonatally with 6-OHDA, but not if rats were co-lesioned as neonates or as adults with 5,7-DHT. The D2 agonist quinpirole induced profound oral activity, but only if rats were lesioned as neonates with 5,7-DHT; or if rats were lesioned with both 6-OHDA (neonatally administered) and 5,7-DHT (adult stage). In all rats lesioned as neonates with 6-OHDA, the 5-HT2 agonist m chlorophenylpiperazine produced enhanced activity, regardless of 5,7-DHT treatment. These findings demonstrate that DA neurons modulate receptor sensitivity status of both DA and 5-HT receptors; and 5-HT neurons do so similarly. This phenomenon is pertinent to animal models of human disorders and in the syndrome spectrum and treatment approach of human neurodegenerative disorders (e.g. parkinsonism, tardive dyskinesia), developmental disorders (e.g. hyperkinetic activity) and psychiatric disorders. PMID- 10389144 TI - Influence of long-lasting administration of neuroleptics on cortical NMDA receptors and phencyclidine-induced deficit in the sensorimotor gating in rats. AB - The aim of this study was to examine the role of cortical NMDA receptors in the antipsychotic action of neuroleptics. Haloperidol (1 mg/kg/day) and clozapine (30 mg/kg/day) were administered to rats in drinking water. Autoradiographic and saturation binding analyses showed that a 3-month treatment with both haloperidol and clozapine increased the density of NMDA receptors labelled with [3H]CGP 39653 (a competitive antagonist) in the parietal and insular cortices. Haloperidol additionally increased the binding of that ligand in the frontal cortex. None of those neuroleptics influenced the binding of [3H]MK-801, an uncompetitive antagonist of NMDA receptors, in the frontal, parietal or insular cortices. A 6 week and a 3-month treatment with haloperidol antagonized the deficit of prepulse inhibition induced by phencyclidine (5 mg/kg s.c.). In contrast, short-term (4 day) administration of that neuroleptic was ineffective. The present study suggests that the increased density of cortical NMDA receptors, induced by long term neuroleptic administration, may overcome the deficit of sensorimotor gating induced by phencyclidine. However, contribution of such an effect to the antipsychotic activity needs to be established. PMID- 10389145 TI - Role of dopamine systems in obsessive-compulsive disorder (OCD): implications from a novel psychostimulant-induced animal model. AB - OCD was once considered a rare psychiatric disorder, but recent studies estimate that, in the general population, the lifetime prevalence of OCD is 1 to 2%, twice that of schizophrenia or panic disorder. The most common form of OCD is compulsive checking. Our studies show that the behavior of rats treated chronically with the dopamine agonist, quinpirole, meets the ethological criteria of compulsive checking in OCD; may have a similar motivational basis as compulsive checking in the human; and is partially attenuated by the anti-OCD drug, clomipramine. Thus, the behavioral changes induced by chronic treatment with quinpirole may constitute an animal model of OCD checking. Since behavioral sensitization is an associated effect of quinpirole treatment, the induction of compulsive checking by quinpirole may involve the same mechanisms as the induction of drug-induced sensitization. In this respect, we demonstrated that the MAO inhibitor clorgyline, not only prevented the development of locomotor sensitization to quinpirole, but also reversed it in sensitized rats. To the extent that the quinpirole treatment is an animal model of OCD with strong face validity, it strengthens the hypothesis that dopamine systems play a role in OCD and raises the possibility that MAO inhibitors, which are used clinically for OCD, may exert their effects by acting at the MAO inhibitor displaceable quinpirole binding site. PMID- 10389147 TI - Influence of fluoroquinolones on the central action of ethanol. AB - The influence of ciprofloxacin, ofloxacin and pefloxacin on acute toxicity of ethanol, ethanol-induced hypothermia, ethanol sleeping time was investigated in mice. Moreover, the combined effect of fluoroquinolones and ethanol on spontaneous locomotor activity, motor coordination in mice and ethanol abstinence syndrome in rats was examined. The fluoroquinolones (20 and 80 mg/kg) were injected intraperitoneally. The drugs were given in single or repeated doses for 7 days. In acute experiments, drugs were given 30 min before ethanol administration. In chronic experiments, the last dose of fluoroquinolones was given 18 h prior to ethanol injection. It has been shown that the fluoroquinolones decrease acute toxicity of ethanol, antagonize its hypothermic effect, decrease ethanol inhibitory effect on motor coordination in mice, and increase ethanol-induced hypermotility in mice and audiogenic seizure response in rats during alcohol abstinence syndrome. Ciprofloxacin and ofloxacin administered repeatedly increase the influence of ethanol on duration of ethanol-induced sleep. The influence of fluoroquinolones on ethanol central action depends on the drug used, its dose and route of administration. PMID- 10389146 TI - Effect of antidepressant drugs on veratridine-evoked glutamate and aspartate release in rat prefrontal cortex. AB - In vivo microdialysis in conscious rats was used to evaluate the effect of local application, through a microdialysis probe, of desipramine (DMI), imipramine and citalopram (CIT), on veratridine-evoked glutamate and aspartate release in rat prefrontal cortex (PFCx). All antidepressant drugs (ADs), given at a concentration of 0.1 mM, significantly inhibited glutamate release, while aspartate release was affected only by DMI and CIT. In contrast, local administration of ADs markedly potentiated veratridine-evoked dopamine and noradrenaline release. Perfusion of clonidine, quinpirole and 1-[3-(trifluoro methyl)phenyl]-piperazine (TFMPP) at 0.1 mM concentration also diminished, evoked release of glutamate and aspartate. The regulation of amino acid release in rat PFCx may be achieved by direct effect of ADs on Na+ channels or indirectly, by involvement of D2/D3, alpha 2 or 5-HT1B heteroceptors activated by the increased level of monoamines in response to the blockade of respective transporters. PMID- 10389148 TI - Further proctolin analogues modified in the position 2 of the peptide chain and their myotropic effects in insects Tenebrio molitor and Schistocerca gregaria. AB - We have extended our studies on the structure-activity relationship in neuropeptide proctolin (Arg-Tyr-Leu-Pro-Thr) by evaluating the effects of a series of proctolin analogues modified in position 2 of the peptide chain, including: [Phe(p-Cl)2]- (1), [D-Phe(p-Cl)2]- (2), [N-Me-Tyr2]- (3), [D-Phe(p NH2)2]- (4), [D-Phe(p-N,N-di-Me)2]- (5), [N-Me-Tyr(OMe)]- (6), [D-3-Pal2]- (7), [L-Nal2]- (8), [D-Nal2]- (9), [Lys(Nic)2]- (10), [D-Lys(Nic)2]- (11), [D-Phe-(p NO2)2]- (12). These peptides were evaluated for myotropic activity on the heart of Tenebrio molitor and contractile activity of the foregut of Schistocerca gregaria. Analogues 1-5, 7-9, and 12 retained a weak cardiotropic activity in Tenebrio molitor while peptides 1, 8 and 12 preserved 15-25% of the locust-gut contracting activity of proctolin. Peptides 2, 4 and 7 showed weak inhibitory activity in Schistocerca gregaria foregut, whereas only peptides 4 and 7 reduced the maximum response to applied proctolin by 64% and 49% respectively, at the 10( 6) M concentration. PMID- 10389150 TI - On some neurochemical aspects of kindling. PMID- 10389151 TI - Glycine/NMDA receptor and seizure phenomena. PMID- 10389149 TI - Stimulatory effect of pituitary adenylate cyclase-activating polypeptide (PACAP) on cyclic AMP formation in the hypothalamus and cerebral cortex of four avians and rat. AB - PACAP38 and PACAP27, tested at 0.0001-1 microM, potently stimulated synthesis of cyclic AMP in the hypothalamus and cerebral cortex of chicks; the effects of vasoactive intestinal polypeptide (VIP) in the two brain regions were much weaker, reaching statistical significance only at 3 microM concentration. This characteristics suggests the existence in the bird's brain of adenylyl cyclase linked PAC1 receptors. PACAP27 (0.001-1 microM) concentration-dependently stimulated cyclic AMP production in the hypothalamus and cerebral cortex of three other birds: duck, goose and turkey; the effects were, however, somewhat lower than those in chicks, but comparable to those found in rats. These data demonstrate PACAP to be capable of potently stimulating cyclic AMP generating system in the avian central nervous system. PMID- 10389152 TI - Nitric oxide and the anticonvulsant action of antiepileptic drugs. PMID- 10389154 TI - Influence of antazoline on the anticonvulsant activity of conventional antiepileptic drugs against maximal electroshock-induced seizures in mice. PMID- 10389155 TI - Anticonvulsive properties of chlormethiazole in mice. PMID- 10389153 TI - Effects of LY 300164, a selective non-competitive antagonist of AMPA/KA receptors, on the protective activity of diazepam and diphenylhydantoin in the kindling model of epilepsy in rats. PMID- 10389156 TI - Influence of antiepileptic drugs on the convulsive and toxic activity of lindane in mice. PMID- 10389157 TI - Influence of prolonged clonic seizures in mice upon their susceptibility to chemical convulsants 2 weeks later. PMID- 10389158 TI - [The role of volume transmission of the adaptogenic signals in the formation of adaptive brain responses]. AB - Different chemical factors of the volume transmission were found to be of importance for the latter process: peptides as possible transporting substances for adaptogenic signals as they were involved in mechanisms of the brain adaptogenic responses, particularly to tetanic stimulation of the neurones and hypoxic stress. Substances secreted by donor-slice cells following tetanic and anoxic stimulation were found to exert an obvious effect upon synaptic transmission inducing a long-term potentiation and protecting it against functional disturbances. Mechanisms of interaction between synaptic and volume transmission signals, are discussed. PMID- 10389159 TI - [Involvement of neurotrophic factors in central mechanisms of behavior in adult animals]. AB - The effect of antibodies against some neurotrophic factors (S-100b protein, CSL, etc.) on elaboration, storage, and retrieval of acoustic startle habituation and concomitant freezing behaviour, was found to be dose-dependent in adult rats. Thus, the 5-mcg dose impaired the consolidation and/or storage process, whereas 2 mcg only impaired the storage of long-term acoustic startle habituation, and 0.5 mcg exerted no effect at all. The habits retrieval displayed no dependence on the antibodies effect. PMID- 10389160 TI - [Study of cytokines in the neural tissue culture]. AB - Natural brain peptides cortexin and epithalamine, and synthetic polypeptides were used to study their effects on organotypic culture of dorsal root ganglia and brain tissue from a 10-11-day old chick embryo. The effects involved activation of the tissue culture. PMID- 10389161 TI - [Transit stage of the long-term facilitation in command neurons of the defensive behavior in sensitized snails]. AB - Sensitising stimuli caused a synaptic facilitation of the Helix lucorum of responses to testing sensory stimulation which lasted over 24 hrs. Following a single stimulus, the facilitation was transient. The same facilitation occurred after 5-HT or cAMP application. Calmodulin and glutamate NMDA-receptor antagonists, as well as the protein synthesis inhibitors suppressed the facilitation in command neurones. The transient synaptic facilitation seems to involve processes of translation/transcription and subsequent synthesis of synapse-specific protein molecules with short life-time. PMID- 10389162 TI - [Possible molecular and cellular mechanisms of gene expression regulation during learning]. AB - The learning plasticity was shown to be accompanied by a serotonin-induced translocation of Rf 0.58 protein, increasing of the G-protein activity, as well as PKA and c-fos gene expression in Helix. The learning marker Rf 0.58 is absent from the juvenile snails. The AP-1 transcription factors in them cannot be determined or induced with serotonin. The level of CRE factors in juvenile snails was comparable with adult and can be regulated by serotonin. The impossibility of sensitisation and avoidance conditioning reflex in juvenile snails can be explain with the AP-1 factors deficit. PMID- 10389163 TI - [Agonistic behavior: model, experiment, perspectives]. AB - Repeated experiences of social victories or defeats in daily agonistic confrontations led to different changes of the brain neurotransmitter activities in male mice with alternative types of social behaviour. Total activation of the brain dopamine metabolism was found in winners. Chronic defeats were accompanied by specific changes in serotonin and noradrenaline metabolism in limbic areas of the brain. Between losers and winners, significant differences were found in emotionality, exploratory activity, locomotion, level of anxiety, communicative behaviour, and alcohol consumption, as well as in immune responses and susceptibility to transplanted Krebs-2 tumour growth, gonadal function, and gastric mucosa damage. A sensory contact technique is proposed for studying the neurophysiological consequences of social conflicts. PMID- 10389164 TI - [Isolation of memory-deficient Drosophila mutants in conditioned courtship suppression paradigm]. AB - Among 33 mutant stocks of Drosophila melanogaster generated by means of P insertional mutagenesis in the system with single P element, 4 stocks have been isolated as demonstrating deficient memory in the conditioned courtship suppression paradigm. Localization of the P insertions never coincided with that of previously known mutations affecting memory. PMID- 10389165 TI - [Proliferative activity and structure-functional organization of chromosomes in cells from the developing brain in respect to genetically determined excitability of the rat nervous system]. AB - Neuroblasts of developing hippocampus of 16-17-day old rat embryo of the line with high threshold excitability are characterised by a high level of proliferative activity and chromosome aberration, as well as high degree of brain chromatin concentration as compared with embryos of a line with low threshold excitability. PMID- 10389166 TI - [Manifestation of fear response in rats genetically predisposed to various kinds of defense behavior]. AB - Acoustic startle response in rats bred for 40 generations of Wistar stock for predisposition to catalepsy, was two-fold higher than in control Wistar rats. In wild Norway rats bred for absence of aggressiveness, the acoustic startle response manifested a three-fold lower amplitude than in aggressive rats. As opposed to tame rats, the startle response reduction in 4 trials was insignificant in wild-type rats. No trend towards reduction of the response was found in cataleptic rats. Genetically determined predisposition to a defensive behaviour seems to be associated with increased manifestation of fear. PMID- 10389167 TI - [Genetic variations of noradrenaline synthesis and reception in the mouse brain and the animal behavior in the new environment]. AB - C57BL mice were found to have the highest locomotion and the lowest emotionality under novel environment out of three strains of mice. Their brain stem TH activity was increased whereas the density of alpha2-ARs and beta-ARs were decreased in their cortex and hypothalamus. The BALB mice were twice as virulent as the CBA mice whereas the emotionality was the same in both strains. In general, low emotionality and high locomotion in novel environment were found in mice with increased activity of norepinephrine synthesis and decreased amount of adrenergic receptors in the brain. PMID- 10389168 TI - [Ethanol modifies the ion selectivity of sodium channels in the rat sensory neurons]. AB - Ethanol was shown to decrease the reversal potential of tetrodotoxin-resistant (TTXr) and TTX-sensitive channels in short-term culture of the dorsal root ganglion cells. The ethanol led to alterations in ionic selectivity of the TTXr channels (its shifting from the X-th Eisenmann selectivity sequence to the XI-th one). The data obtained suggest that the findings were due to selectivity filter modification because of disturbed hydrogen bounds in the channel macromolecule. PMID- 10389169 TI - [Stimulus-dependent effects of the sodium channel blockers in the C-fiber sensory units]. AB - The data obtained suggest a use-dependent inhibition in the skin terminals of the C-fibre sensory units. The terminals are discussed in respect to search of efficient local anaesthetising agents as well as cardiac anti-arrhythmic agents with obvious neurotropic effects. PMID- 10389170 TI - [Effect of the cytoskeleton dynamic condition on the neuronal plasticity]. AB - Infringement of the Lymnaea stagnalis cytoskeleton condition affected preservation and repeated development of plastic responses. Stabilising of the microtubules led to a dependence of the development and preservation dynamics of the plastic responses. Stabilising of the microfilaments transformed short-term plastic responses into long-term ones. The findings suggest a key role of reorganisation of the cytoskeleton in neuronal plasticity. PMID- 10389171 TI - [Involvement of the intracellular regulatory systems in the adaptive effect of short-term anoxia in vitro]. AB - Moderate long-term activation of intracellular regulatory system (IRS) was found to be manifested as an increase in the bound calcium content and intensity of the phosphoinositide metabolism following a 30-minute re-oxygenation in the rat olfactory cortex perfused slices. The perfusate induced a similar activation in intact slices-recipients. Long-term anoxia induced a biphasic NMDA-dependent increase in intracellular free Ca2+ and pathogenic hyperactivity of the IRS. The pathogenic events could be prevented by preconditioning of the slices by either short-term anoxia (STA) or post-STA donor perfusate. PMID- 10389172 TI - [Modulatory effects of various factors of muscular origin on the function of the motor nerve endings]. AB - Muscle incubate was shown to contain factors capable to increase the transmitter release in low-effective synapses and decrease it in high-effective those, dueto activation and inhibition, respectively, of the processes governing formation of available store of the transmitter. Activatiory effect of the low-molecular fraction on the low-effective synapses was correlated with the concentration of histidine-containing substances in this fraction. PMID- 10389173 TI - Molecular complexity at the synapse: new proteins and multiple isoforms detected in Drosophila. AB - Synaptic transmission and its context-dependent modification are pivotal for all forms of information processing in the nervous system including learning and memory. The molecular components of the presynaptic nerve terminal are therefore under intensive study. Using a reverse genetic approach in Drosophila we have cloned the first invertebrate homologue to vertebrate synapsins and identified two new protein families, the "synapse-associated protein of 47 kD" (SAP47) and the cysteine string proteins (CSPs), which are conserved throughout higher eukaryotic evolution. Information on the molecular, cellular and systemic functions of these proteins and their isoforms is summarized in this review. PMID- 10389175 TI - Experience-dependent development of the adult optic lobe and central brain in Drosophila melanogaster. AB - The optic lobes, mushroom bodies (MB) and central complex (CC) of Drosophila melanogaster were investigated in order to find out whether rearing in different light regimes affect their size. Flies raised in constant light up for up to four days post eclosion had a lamina that was about 30% larger than in flies kept in constant darkness. A volume difference between light- and dark-reared flies could also be observed for the lobula plate, the MBs, and the CC. When the flies were kept in the dark for the first 12 hours of their adult life and then brought back to constant light for the next 3.5 days, the lamina was as small as the laminae of flies raised for four days in constant darkness. This finding suggests a critical period for lamina development during day one of the imago. Mutant studies suggest that the molecular mechanisms underlying this experience dependent development might be quite diverse. For example, the structural plasticity of the mushroom bodies is abolished in the dunce mutation, whereas the light-dependent growth of the lamina is not. Finally, studies of optomotor behavior indicate an adaptational role for the structural plasticity in the optic lobe. Surprisingly, dark-reared flies see better under low light conditions than their light-reared counterparts. This suggests that a small lamina is not a bad lamina in the sense that dark-reared animals see worse. They rather adapt to the specific light-conditions they were growing up in. PMID- 10389174 TI - Age-dependent changes in memory and mushroom bodies in the Drosophila mutant vermilion deficient in the kynurenine pathway of tryptophan metabolism. AB - Evolutionary conservation of homologous genes that cause related phenotypes in humans and Drosophila help to unravel genes implicated in polygenic human diseases. Among them are neurodegenerative disorders, such as Huntington, Parkinson, Alzheimer and HIV-induced diseases. They are characterized by a late onset disturbances of memory, changes in volumetric indices of the brain structures involved in memory formation, synaptic and glial pathology, and altered content of the intermediates of the kynurenine pathway, the endogenous modulators of the NMDA receptors. This pathway in conserved in insects, rodents and humans. We, therefore, studied the effects of aberrant tryptophan metabolism on memory, brain plasticity, synaptic and glial immunoreactivity in the Drosophila mutants vermilion (no kynurenines) and cinnabar (excess of neuroprotective kynurenic acid) over the life time. The mutant vermilion demonstrated gradual decline of 3-th memory performance and complete memory failure on the 28th day of life in a paradigm of conditioned courtship suppression. A drastic increase in the volume of the calyces of the mushroom bodies, and a decay in immunochemical staining of this brain structure with antibodies to synaptic protein csp and glia, precede the age-dependent memory defect and develop from the 12th day of adult life. PMID- 10389177 TI - Transduction and transmission properties of primary nociceptive afferents. AB - The prototypical primary nociceptive afferent is the polymodal C-fiber nociceptor, which responds to noxious thermal, mechanical, and chemical stimuli. C-fiber nociceptors are peripheral terminals of small neurons in the dorsal root ganglia (DRG). DRG neurons must therefore supply their peripheral terminals with the molecular machinery for the encoding of noxious stimuli into trains of action potentials. The following phenomena are known for this encoding process in vivo: 1) adaptation: for a constant stimulus intensity the action potential discharge decreases slowly within 2-3 seconds, 2) fatigue: recovery from adaptation may take ten minutes or more, 3) sensitization: preceding tissue damage enhances the response, particularly to heat stimuli. Recent studies in vitro have provided important clues about the molecular mechanisms underlying these phenomena. Several membrane receptors and channels are specifically expressed in small nociceptive neurons, such as vanilloid receptors (VR1), purinergic receptors (P2X3), acid sensing ion channels (ASIC), and TTX-resistant Na-channels. In the near future, we may therefore expect major advances in our understanding of the transduction of noxious stimuli into generator potentials and transformation into trains of action potentials. Along the axon that leads from the innervated tissue to the spinal cord, primary nociceptive afferents have a limited capacity to transmit high impulse rates, suggesting a different composition of voltage-gated channels than in other primary afferents (low-threshold mechanoreceptors and thermoreceptors). Finally, the DRG neuron also supplies its central terminals with the molecular machinery for synaptic transmission and its presynaptic modulation. Progress in understanding the cellular mechanisms at both ends of the primary nociceptive neuron promises to lead to new analgesic treatment modalities for both acute and chronic pain. PMID- 10389176 TI - Ionic mechanisms underlying TRH-induced prolactin secretion in rat lactotrophs. AB - Whole cell patch-clamp experiments were performed in clonal rat pituitary cells (GH3/B6 cells) to further analyze an inward-rectifying K current (IK, IR) which is suggested to be involved in the thyrotropin-releasing hormone (TRH)-induced increase in prolactin secretion from these cells. Using the class III antiarrhythmic agent E-4031 which is known as specific blocker of ether-a-go-go related gene (ERG) K channels, the inward-rectifying K current could be isolated as the drug-sensitive current. To elucidate in molecular basis of this current, comparative experiments were performed in CHO cells which served as heterologous expression system for RERG, the rat homologue of the human ERG (HERG). It is shown that the inward-rectifying K current has properties identical to those mediated by channels encoded by RERG. In external 5 mM K+ solution, the ERG-like current IK, IR was an outward current in the physiological potential range, and this outward current could be strongly reduced by TRH. A specific block of IK, IR was able to mimick the second phase of the TRH-response which is characterized by a depolarization and/or by an increase in the frequency of Ca action potentials. These data show, that the ERG-like current in GH3/B6 cells contributes to the maintainance of the resting membrane potential and that it plays an important part in the mechanisms of the effects of TRH leading to an increase in prolactin secretion. PMID- 10389178 TI - Role of Ca2+ release from ryanodine stores in generation of NMDA-induced Ca2+ signal in rabbit hippocampus. AB - In vivo microdialysis combined with measurements of 45Ca efflux from pre-labelled rat hippocampus has been utilised in our laboratory to demonstrate NMDA-evoked 45Ca2+ release to dialysate, reflecting calcium-induced calcium release (CICR) via ryanodine receptors (RyR). In the present study we attempted to reproduce this phenomenon in the rabbit hippocampus. Application of 1 mM NMDA to dialysis medium induced a decrease in Ca2+ concentration in dialysate, as a result of extracellular Ca2+ influx to neurones. The release of 45Ca2+ was not observed, instead a decrease in 45Ca2+ efflux rate from the NMDA treated rabbit hippocampus was noted, along with release to dialysate of prostaglandin D2, taurine and phosphoethanolamine. All these effects, reflecting different steps of intracellular calcium signalling, were insensitive to 100 microM dantrolene and 50 microM ryanodine, RyR modulators known to interfere with NMDA-evoked 45Ca2+ release in the rat hippocampus. Thus, although the results of this study demonstrate the role of extracellular Ca2+ influx to neurones in NMDA-evoked generation of Ca2+ signal in the rabbit hippocampus, the activity of CICR was not detected. PMID- 10389179 TI - [Morphine decreases the voltage sensitivity of the slow sodium channels]. AB - Morphine was shown to decrease in a dose-dependent manner the effective charge transfer in tetrodotoxin-resistant (TTXr) sodium channel activation system in short-term cultured dorsal root ganglion cells. Morphine seems to interact with opioid receptors because of total block of the binding by naloxone and naltrexone. Neither activating, nor inhibiting G-protein agents exerted any effect on this process. The morphine signal was blocked by extracellular application of 2 x 10(-4) M ouabain. The findings suggest existence of sodium signalling pathway involving receptors, Na+, K(+)-ATPase and the TTXr sodium channels. PMID- 10389180 TI - [Effect of cyclic AMP on the excitation and response of command neurons responsible for the snail defensive behavior caused by sensory stimulation]. AB - Extraneuronal application of db-cAMP or intraneuronal injection of cAMP were found to increase the neural membrane excitability and synaptic facilitation in neural responses to sensory stimulation of Lpl1 and Rpl1 neurones. The db-cAMP exerted no effects on neural responses to tactile stimulation of the snail foot or mantle. The intraneuronal injection of cAMP produced synaptic facilitation only in neural responses to quinine application to the snail head. The findings suggest the cAMP selective involvement in postsynaptic mechanisms of inducing the long-term facilitation transient stage. PMID- 10389181 TI - [Effect of the ion composition of the calcium-free medium and cardiomyocyte damage during "calcium paradox" in rats]. AB - The findings reveal that the degree of myocardial damage in the "calcium paradox" does not depend on the contracture strength, that the contracture attenuation due to a decreased concentration in the Ca-free medium is not equal to the cardiocytes protection as compared with other means. Mg2+ and the studied means of myocardial protection seem to play a major role in assessment of the "calcium paradox" development and explain the difference in results of the hyposodium medium effects in the "calcium paradox". PMID- 10389182 TI - [Dependence of myocardial damage on the anion composition and osmotic pressure in the extracellular fluid during "calcium paradox" in rats]. AB - The data obtained reveal that elevation of extracellular osmolarity with sucrose during reintroduction of Ca-containing medium after 10 minutes of Ca2+ removal prevents loss of haemoglobin in a concentration-dependent mode. Reducing the extracellular osmolarity of the reperfusion medium by means of decreasing the concentration of sodium chloride and calcium chloride exacerbates the loss of haemoglobin from the cardiomyocytes. There is a close correlation between the water contents in tissues and the loss of haemoglobin during the "calcium paradox". The findings suggest dependence of the heart damage during the "calcium paradox" on anionic composition of extracellular space and activity of anionic transporters. PMID- 10389183 TI - [Effect of long-term cold exposure on apoprotein A-I, B, and E levels in the rat serum]. AB - An increase in the quantity of triglyceride-rich VLDL as well as the total fraction VLDL and LDL occurred following the cold adaptation. The contents of apoB and apoE, respectively, increased. However, a 5-15-day exposure to cold decreased the LDL2 and apoA-1 content. In 49 days of the experiment, the LDL2 and apoA-1 content was restored. PMID- 10389184 TI - [Study of the mutagenic and antimutagenic activity of the human and animal blood]. AB - Mutagenic and antimutagenic properties of humans, bull, rabbit, chick, rat, and mouse were studied using seeds of Crepis capillaries (the chromosome aberration test). Antimutagenic activity was found in pre-processing, combined, and post processing conditions of the seeds with a blood relative to the mutation inductors. Whole human and animal blood was found to be capable of inducing chromosome aberrations far exceeding the level of spontaneous mutations. Dilution of the blood did not eliminate its antimutagenic activity. PMID- 10389185 TI - [Study of the activity of receptors with nonmyelinated fibers during heat exposure of the hairy cat skin]. AB - A nerve activity in heating of the cat hairy skin was studied in acute experiments in order to find out if the C-receptors take part in peripheral coding of heat information. An integrative cross-correlation technique was used to analyse neurograms in the real time. No C-afferent activity was found in the thermal stimulation up to 41 degrees C of the hairy skin. PMID- 10389186 TI - [Effect of mediodorsal thalamic nucleus on the respiratory neurons from medulla oblongata and the rat respiration during hypoxia]. AB - In the normal as well as in the oxygen deficiency conditions the research has been conducted to study the influence of associative mediodorsal (MD) nucleus of thalamus on impulsive activity of respiratory neurons of medulla oblongata of respiration. In conditions of normal atmospheric pressure, before the uplift of the animals, the electrical stimulation of MD of nucleus of thalamus has had mainly inhibiting influence. In the initial phase, on 4-5 thousand meter altitude, activation of frequent discharge of neurons occurred, the respiration has become frequent as well. In this situation the inhibiting influence of stimulation of MD nucleus of thalamus was more accentuated than in conditions of normoxia. In the second phase, 7.5-8 thousand meters, the opposite occurred, i.e. reduction of respiratory center activity of medulla oblongata and thalamus. In this difficult conditions of hypoxia, a reduction of impulsive activity of neurons has been observed; the respiration was becoming slower and surface. Meanwhile, the inhibiting influence of thalamus was not significant. PMID- 10389187 TI - [Morphometric analysis of the effects of vasopressin in the rat kidney collecting tubules]. AB - A significant increase in the water permeability was found in the rat outer medullary collecting duct (OMCD) cells in presence of 10-7M of vasopressin. The latter caused a decrease in the OMCD cell volume in isoosmotic medium in adult rats. In pups, the water permeability of the OMCD cells was very high. Vasopressin seems to be unable to decrease the cell volume of the OMCD cells in pups which suggests an immaturity of the cell transduction mechanism. PMID- 10389188 TI - [Changes in the level of specific proteins in the rat kidney during long-term dehydration]. AB - A significant amount of specific proteins are involved in kidney's response to vasopressin. A relative content of 120 kDa protein in the Wistar rat kidney medulla following a 3-day dehydration, was found to be 0.975 +/- 0.037 and after drinking 4% sucrose solution alone--0.871 +/- 0.038. The difference of protein 120 kDa content was not revealed in Brattleboro rats: in the control rats it was 0.814 +/- 0.044, and in the dehydrated ones--0.854 +/- 0.020. The findings suggest that vasopressin directly regulates the 120 kDa protein level in the kidney inner medulla. PMID- 10389189 TI - [The role of zinc in the iliac absorption of various L-tryptophane forms in chickens]. AB - Zinc was found to enhance absorption of free and "peptide" L-tryptophan across the chick mucosal brush border. Intestinal absorption of free L-tryptophan can be partially inhibited by excessive amount of zinc. The findings suggest that zinc interacts with free and "peptide" L-tryptophan transports protecting them against degradation. PMID- 10389190 TI - [The condition of the intestinal mucosa of the proximal digestive tract subjected sectioning of the jejunal wall]. AB - The balance between secretion and degradation of the mucosa glycoproteins in the dog intestine is preserved with the aid of local reflexes in both serous-muscular and mucous-submucous layers of the wall. Irrespective of major alterations in the glycoproteins synthesis in the mucosa, protective properties of the submucous layer are maintained owing to a high content of acetylneuraminic acid and low level of mucous glycoproteins degradation. PMID- 10389192 TI - [Discovery of enterokinase (1899-1999)]. PMID- 10389191 TI - [Effect of prednisolone on the gastric juice basal secretion during blockade of M cholinoreceptors by gastrozepin in rats]. AB - A moderate depression of the stomach M-cholinic receptors was found to facilitate the prednisolone glucocorticoid stimulating effect on basal gastric secretion. PMID- 10389193 TI - [A creative journey of Nikolai Petrovich Shepoval'nikov (1872-1945)]. PMID- 10389194 TI - [Liudmila Nikolaevna Ivanova]. PMID- 10389196 TI - Plaque pathology and coronary thrombosis in the pathogenesis of acute coronary syndromes. AB - Coronary atherosclerosis is by far the most frequent cause of ischemic heart disease and plaque disruption with superimposed thrombosis is the main cause of the acute coronary syndromes of unstable angina, myocardial infarction, and sudden coronary death. Therefore, for event-free survival, the vital question is not why atherosclerosis develops but rather why, after years of indolent growth, it suddenly becomes complicated by life-threatening thrombosis. Therefore, we have to focus on plaque composition and vulnerability to rupture and plaque thrombogenicity rather than on plaque size and stenosis severity. The risk for plaque disruption depends more on plaque vulnerability (plaque type) than on degree of stenosis (plaque size). Lipid-rich and soft plaques are more vulnerable and prone to rupture than collagen-rich and hard plaques. They are also highly thrombogenic after disruption because of high content of tissue factor. There seems to be three major determinants of a plaque's vulnerability to rupture: 1) the size and consistency of the lipid-rich atheromatous core, 2) the thickness of the fibrous cap covering the core, and 3) ongoing inflammation and repair processes within the fibrous cap. Lipid accumulation, cap thinning, lack of smooth muscle cells (smc), and macrophage-related inflammation destabilize plaques, making them vulnerable to rupture. In contrast, smc-related healing and repair processes stabilize plaques, protecting them against disruption. Plaque size or stenosis severity tell nothing about a plaque's vulnerability. Many vulnerable plaques are invisible angiographically due to their small size and compensatory vascular remodeling. PMID- 10389197 TI - Role of inflammation in the pathogenesis of unstable coronary artery diseases. AB - Inflammation has been shown to play a pivotal role in ischemic heart disease, in particular unstable angina. The instability that characterizes this syndrome is related to the waxing and waning of ischemic stimuli, especially thrombotic ones. Angiographically and autoptically the severity of the atherosclerotic background in unstable angina does not differ from that in chronic stable angina, but in the former mural thrombi are often found and coronary atherosclerotic plaques are characterized by an inflammatory infiltrate, mostly consisting of activated lymphocytes, macrophages and mast-cells. In addition to these local findings, systemic evidence also suggests the importance of the role of inflammation in unstable angina as platelets, neutrophils and monocytes are activated, and elevated levels of serum markers of inflammation, e.g. C-Reactive Protein, have been consistently found. CRP has been demonstrated to be a reliable marker of prognosis in coronary heart disease. The consequences of inflammation are a disruption in the dynamic balance between antithrombotic and prothrombotic activities, an altered extracellular matrix metabolism, hyper-reactivity of cells such as monocytes and smooth muscle cells, all important features of unstable angina. These findings have important prognostic implications, since markers of inflammation are associated to a worse prognosis, and may also have therapeutic implications in the near future. PMID- 10389198 TI - Predictive value of fibrinolytic factors in coronary heart disease. AB - An impaired fibrinolytic function, as evidenced by increased plasma concentrations of PAI-1, tPA antigen and tPA/PAI-1 complex, or a decreased capacity to release active tPA on exercise, is more common in individuals suffering from myocardial infarction. These factors, especially the tPA/PA-1 complex, also predict myocardial infarction in patients with manifest coronary heart disease, such as angina pectoris or a previous myocardial infarction. There is a highly statistically significant correlation between tPA/PAI-1 complex and both PAI-1 and tPA antigen. It is important to test these factors in prospective studies on healthy individuals. PMID- 10389199 TI - A new method of measuring C-reactive protein, with a low limit of detection, suitable for risk assessment of coronary heart disease. AB - A new indication has been proposed for C-reactive protein (CRP) as a prognostic risk marker of coronary heart disease (CHD). The new indication calls for accurate (true and precise) measurement of CRP within the conventional reference range (< 5 mg/L). The existing turbidimetric and nephelometric methods do not cover the required measuring range and thus time-consuming and labour-intensive enzyme immunoassays have been used for the clinical studies focusing on CHD risk. We developed a new method based on microparticle enhanced turbidimetry, which attained the required limit of detection, while keeping the upper measuring limit comparable to the existing turbidimetric and nephelometric methods. The superior characteristics of the new method were realised by mixing two types of microparticle reagents differing in microparticle size and reactivity of coated antibody. The analytical detection limit of the method was 0.28 mg/L with use of only 2.5 microliters serum. The method showed good precision at 2 to 3 mg/L, the critical concentration range for CHD risk assessment. Other performance data including dilution linearity, method comparison, and interference study also met the requirements for the practical use in the clinical laboratories. Sera from 354 apparently healthy blood donors were measured in a reference range study. The reference range estimated after log-transformation was 0.16 mg/L to 7.57 mg/L CRP, with a total range of 0.09 mg/L to 21.0 mg/L. The distribution of CRP concentrations in this population was comparable to other results that established the use of CRP as a risk marker of CHD in a prospective study. PMID- 10389200 TI - Strategies for clinical assessment of patients with suspected acute coronary syndromes. AB - Clinical assessment of patients with acute coronary syndromes is routinely employed for risk stratification and selection of treatment strategies. Both, patients risk and treatment options vary in unstable angina, non-Q-wave infarction and massive Q-wave infarction. Clinical symptoms and admission ECG are the key elements in the daily work-up of chest pain patients. However, absent or nonconclusive ECG changes, even in patients with confirmed acute myocardial infarction, and high variability of clinical symptoms, particularly in elderly patients, limit their diagnostic value and their precision for risk stratification of acute coronary syndromes. Additional diagnostic testing such as 24 hours Holter ECG depicting prolonged episodes of ST-segment depression, and radio-nuclide tomography using Tc-Sestamibi improves accuracy of risk assessment. However, in clinical practise these techniques are not readily available. Troponins, particularly cardiac specific troponin T, are paramount for risk stratification of acute coronary syndromes, either alone, or in combination with admission ECG or a predischarge exercise stress test. Stratifying individuals into high, intermediate, and low risk for death and subsequent cardiac events may aid to improve outcomes by tailored use of a more aggressive therapy in these subjects. PMID- 10389201 TI - Therapeutic implications of the use of cardiac markers in acute coronary syndromes. AB - Patients admitted with suspicion of an acute coronary syndrome still constitute a diagnostic, prognostic and therapeutic challenge. In the vast majority of these patients the diagnosis is dependent on the results of measurements of cardiac markers. The troponins have a higher diagnostic sensitivity and specificity for detection of myocardial damage than previous markers and have also been proved to be valuable for early risk stratification. There is emerging evidence that these new cardiac markers might be used for selection of treatment in various forms of acute coronary syndromes. PMID- 10389202 TI - The use of cardiac markers in acute coronary syndromes. AB - Cardiac markers can be used effectively for diagnosing, triaging, treating and monitoring of patients with acute coronary syndromes. Therefore, in accord with the recommendation of the NACB and IFCC, the use of myoglobin and troponins, especially troponin T (TnT) because of the broader data base, are well-suited to fulfil the needs of management of acute coronary syndromes. The present paper reviews recent data to evaluate their relevance, especially in deciding the treatment of patients with non-ST elevation acute coronary syndromes. Furthermore, economic studies show that the use of even the more expensive diagnostics is associated with tremendous cost savings due to more efficient patient management. The use of cardiac markers therefore represents a valuable tool in the management of these patients. The best combination is that of myoglobin with troponin. PMID- 10389203 TI - Biochemical markers of thrombolytic success. IFCC Committee on Standardization of Markers of Cardiac Damage. AB - This paper reviews the clinical criteria necessary for use of thrombolytic drug therapy, the role of monitoring biochemical markers for detection of successful reperfusion following thrombolytic therapy, and the role of monitoring markers following percutaneous intervention to assess risk. Review of several studies demonstrates that early monitoring of myoglobin, cardiac troponin I, cardiac troponin T, and CK MB mass provides greater than 80% sensitivity and specificity for detecting reperfusion within 90 minutes following the initiation of therapy. However, the number of acute myocardial infarction patients studied are small. Since no prospective studies are reported, additional studies are necessary prior to clinical acceptance of routine monitoring of markers for detection of reperfusion. Most important, TIMI 3 flow patients cannot be differentiated from TIMI 2 flow patients. Monitoring markers after PTCA does appear to assist in short term (30 day) risk stratification. However, more studies are needed to assess the use of cardiac troponins. PMID- 10389204 TI - The need for a point of care testing: an evidence-based appraisal. AB - The need for point of care testing (POCT) must be seen within the context of the need for biochemical diagnosis of patients suspected of acute coronary syndromes (ACS). The electrocardiogram is the initial test to select patients for thrombolysis and risk stratification. The majority of patients presenting with chest pain do not have AMI and diagnostic sensitivity of the ECG is only 55-75%. This means that biochemical diagnosis to confirm or exclude a diagnosis of acute myocardial infarction (AMI) is required for 90% of patients who present with suspected ACS. This can be achieved using myoglobin, creatine kinase and its MB isoenzyme and the cardiac troponins, cardiac troponin T and cardiac troponin I within 4-12 hours of admission, depending on the markers selected. Measurement of the cardiac troponins is superior to conventional tests as it allows risk stratification in patients with unstable angina in addition to a definitive diagnosis of AMI. Rapid diagnoses can have direct influence on length of stay and admission and discharge policies. The rationale for POCT is the improvement in analytical turnaround time (TAT). A number of systems for POCT have been developed ranging from conventional analysers used next to the patient to whole blood systems using immunochromatography. Laboratory and clinical evaluations of these systems have shown that they are accurate, equivalent to laboratory based diagnosis and can be related to outcome. In a prospective randomised controlled trial of POCT compared to laboratory testing we have shown improved TAT (20 minutes vs. 72 minutes) and significant reduction in hospital stay in patients randomised to POCT. Rapid diagnosis is effective and cost effective for patient management but only within the context of data driven decision-making protocols. The use of POCT will depend on the need for and the ability to achieve a rapid TAT for clinical decision making. POCT for cardiac markers has a definite place in management when very short TAT is required such as the ED. If the laboratory TAT exceeds 25% of the decision time, then POCT will be required. If therapeutic decisions are to be made, only POCT will be able to fulfil the TAT requirement. PMID- 10389205 TI - Biochemical markers of cardiac damage: from traditional enzymes to cardiac specific proteins. IFCC Subcommittee on Standardization of Cardiac Markers (S SCM). AB - Measurement of cardiac markers in blood has been the mainstay for diagnosis of acute myocardial infarction for nearly 50 years. The field has evolved from measurement of enzyme activity to mass concentrations of proteins using automated non-isotopic immunoassays. With changing clinical practices, cardiac markers are now needed to detect the presence of minor myocardial infarction in patients with unstable angina. Outcome studies have shown that patients with increased troponin are at high short-term risk for death and AMI. Recent developments involve the use of cardiac markers to select the most appropriate therapy for patients with acute coronary syndromes. The success of new cardiac markers such as troponin is due to their high cardiac specificity and the existence of assays with low detection limits. Traditional enzymes such as CK and CK-MB are thought to be released only in situations of irreversible myocardial necrosis. In the case of cardiac troponin, clinical observations and animal studies suggest that cytosolic free troponin may be released in reversible ischemia in addition to irreversible cell damage. The IFCC S-SCM has recommended use of two cut-off concentrations for cardiac troponin to differentiate normal from minor myocardial injury and AMI. A low cut-off may detect reversible ischemic events in some cases. PMID- 10389207 TI - Evidence based approach to practice guides and decision thresholds for cardiac markers. AB - There is substantial evidence for use of CK-MB, myoglobin, cardiac troponin T (cTnT) and cardiac troponin I (cTnI) for diagnosis of myocardial infarction (MI) and risk stratification of the acute coronary syndromes. Timing of specimen collection, patient population, and decision limits are important considerations. Serial CK-MB measurement must be considered a benchmark of cardiac markers. The National Heart Attack Alert Program (NHAAP) determined that high quality clinical trials show that serial CK-MB measurements are very accurate for diagnosis of MI and have large clinical impact. Meta-analysis of CK-MB mass for retrospective MI diagnosis yielded a clinical sensitivity of 96.8% and a specificity of 89.6% for sampling at 12-48 hours. Numerous studies of myoglobin have demonstrated a high negative predictive value and high sensitivity 2-6 hours after presentation. The NHAAP group, however, rated performance of myoglobin as modestly accurate with small clinical impact. cTnI has a clinical sensitivity and specificity for diagnosis of MI in the range of 90% and 97%, respectively. However decision limits for cTnI may differ by up to 10-fold for different assays, indicating need for standardization. Meta-analysis showed that cTnT had a sensitivity of 98.2% (CI: 97-99%) for diagnosis of MI with lower specificity due to detection of minor myocardial injury. Both cTnT and cTnI are more useful for stratifying risk in acute coronary syndrome patients than CK-MB mass. The predictive ability of both cTnT and cTnI is optimized by sampling > 6-12 hours after symptoms onset. But in unstable angina, the odds ratios for predicting outcomes of MI/short-term mortality with 12-24 hour sampling for cTnT was 2.7 the odds ratio for cTnI did not differ significantly at 4.2. PMID- 10389206 TI - Can troponin T replace CK MBmass as "gold standard" for acute myocardial infarction ("AMI")? AB - Diagnosis of definite "Acute Myocardial Infarction" by the old WHO criteria depends on the diagnostic sensitivity and specificity of the biochemical marker used. Troponins have higher diagnostic sensitivity and specificity than the current "gold standard" for AMI, CK MBmass. Troponin T (TnT) provides both diagnostic and prognostic information on Minor Myocardial Damage (MMD) even in patients without increases of CK MBmass. Consequently, we evaluated the possibility of replacing CK MBmass with TnT. We first re-evaluated a previous, well-documented material of 502 time series from AMI-suspected cases, 50% of which were primarily classified as AMI by CK MBmass > or = 10 micrograms/L. We found that a TnT discriminator limit of 0.40 microgram/L gave the same AMI prevalence. We then identified from our laboratory data base 1995-1998 acute patient episodes with > or = 3 pairs of CK MBmass and TnT. This resulted in 754 episodes with max CK MBmass > or = 10 micrograms/L (AMI), 93 episodes with maximal CK MBmass < 10 micrograms/L and TnT > or = 0.10 microgram/L (MMD), and 730 episodes with max concentrations below the discriminators of both markers (NOT-MMD). TnT > or = 0.40 microgram/L detected 91% of all AMI giving a posterior probability of "AMI" > 99%. The criterion: "maximal TnT within the interval 0.10 0.40 microgram/L" detected 94% of all MMD and 9% of all AMI giving posterior probabilities about half MMD and half "small AMI", the latter characterized by less than 3-fold increased maximal CK MBmass. Thus, this TnT interval confirmed the gradual transition between MMD and small AMI. We suggest gradation of myocardial damage by TnT. PMID- 10389208 TI - Proposals from IFCC Committee on Standardization of Markers of Cardiac Damage (C SMCD): recommendations on use of biochemical markers of cardiac damage in acute coronary syndromes. AB - This paper presents evidence and suggestions from the IFCC C-SMCD on the use of biochemical markers for the triage diagnosis of acute coronary syndromes. There is general agreement that both 'early' and 'definitive' biochemical markers are necessary and that these assays must be available with a turnaround time of 1 h or less. Currently, myoglobin is the marker that most effectively fits the role as an 'early' marker, whereas 'definitive' markers are cardiac troponins. Since the sensitivity of the initial electrocardiogram is only 50% for detecting myocardial infarction, the use of biochemical markers may significantly contribute to the early diagnosis. New sensitive biochemical markers, particularly the cardiac troponins, are presently the best criterion to detect the presence of small myocardial cell damage. Two decision limits are probably needed for the optimum use of troponins: a low abnormal value suggesting the presence of myocardial damage and a higher value suggesting the diagnosis of myocardial infarction. Additional studies should be performed to establish limits for each commercially available assay. Finally, it is recognized that there is no need for the use of any biochemical marker when the clinical diagnosis is unequivocal, other than for diagnosing reinfarction, estimating the infarct size, and monitoring thrombolytic therapy. PMID- 10389209 TI - Proposals from the IFCC Committee on Standardization of Markers of Cardiac Damage (C-SMCD): strategies and concepts on standardization of cardiac marker assays. AB - The search for sensitive and specific biochemical markers of cardiac damage has resulted in development of methods for the measurement of cardiac markers such as myoglobin, CK-MB mass and the cardiac troponins (cardiac troponin I and cardiac troponin T). There have been new clinical applications of already known markers based on improved, reformulated methods which often depend on advanced technologies. These developments are connected with analytical and interpretative challenges for the laboratory manager and for the clinician. In this situation, it is essential that international professional societies develop comprehensive and carefully elaborated guidelines for the quality management and use of these measurements and their results. Several professional associations have formed committees working on different issues regarding the measurement of biochemical markers of cardiac damage. Recognizing the huge interest in this field and substantial diagnostic relevance of these markers, the IFCC has established the Committee on "Standardization of Markers of Cardiac Damage" (C-SMCD) in 1997 inviting the already operating American and European groups to designate some of their members into the new committee. The task of the IFCC C-SMCD is to coordinate the different activities of the national groups, with preparation of international documents and recommendations under the auspices of IFCC and to initiate various standardization activities. The establishment of consensus/reference methods as well as development of primary and secondary matrix reference materials for markers of cardiac damage are extremely important in order to achieve comparability of test results, thus leading to an effective patient care. PMID- 10389210 TI - New approach to standardisation of human cardiac troponin I (cTnI). AB - Six different cTnI assays (5 commercial and one assay under development) were run with either their own calibrators or with standard dilutions of different forms of cTnI (free or complexed). Twenty-one serum samples from 7 AMI patients were analysed using these combinations of calibrators and assay constructs. The range of cTnI concentrations as measured in AMI serum samples by different assays was more than 10-fold using each assays own standards or when using free native or recombinant cTnI. With recombinant or in vitro formed ternary cTnI-cTnT-TnC complexes the differences were smaller (3-fold). The lowest between-manufacturer bias (1.4-fold) was obtained when the heart tissue derived native troponin complex was used for the calibration of the assays. For all assays, calibrated against this troponin I complex, the recalculated cut-off values were within 0.1 0.25 microgram/L. We conclude that to reduce assay-to-assay variation, native troponin complex is presently the preferred alternative for the standardisation of cTnI assays. PMID- 10389211 TI - Use of recombinant human cardiac Troponin T for standardization of third generation Troponin T methods. AB - The Elecsys Troponin T third generation assay uses recombinant human cardiac troponin T as standard material. The new assay has a linear calibration curve. Thus, linearity problems observed with the second generation assay have been eliminated. The assay has high precision, especially at the low end of measuring range (inter-assay CV < 10% at 0.1 microgram/L). The new standardization does not change the cut-off value of 0.1 microgram/L. The use of recombinant human cardiac troponin T as standard material enables a reproducible and reliable standardization of troponin T assays. PMID- 10389212 TI - Natriuretic peptides in assessment of left-ventricular dysfunction. AB - The heart secrets two different natriuretic peptides, atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP), which have potent vasorelaxant, diuretic, and natriuretic actions. They are main tools in the body's defense against volume overload and hypertension. The natriuretic peptides (NP) are synthetized as prohormones. The C-terminal endocrinological active peptides and their N-terminal prohormone fragments are found in plasma. The NP system is maximally activated in ventricular dysfunction. However, NP:s are also increased in patients with renal failure or pulmonary hypertension, and increases may be found in arterial hypertension or liver cirrhosis. Among all NP and prohormone fragments currently BNP is the most promising candidate analyte for routine diagnosis. BNP is also superior to other neurohormones for diagnosis of left ventricular dysfunction (LVD) or estimating prognosis in LVD or during the subacute phase of myocardial infarction. For primary care physicians BNP measurement is useful to decide which patient with suspected heart failure warrants further investigation, particularly when assessment of left ventricular function is not readily available. BNP has an excellent negative predictive value particularly in high risk patients. For the cardiologists the NP:s are helpful for monitoring therapy and disease course in LVD patients and for estimating prognosis in LVD and myocardial infarction patients. There is now sufficient evidence to encourage physicians to gain experience with NP as a supplement in the diagnosis of patients suspected of having heart failure. An increase in BNP is serious enough to warrant follow-up examinations. PMID- 10389213 TI - Albuminuria in ischemic heart disease. AB - Proteinuria associated with acute heart disease was studied prospectively in 160 patients admitted to the coronary care unit with suspected AMI. Series 1 comprised 150 patients, divided into the following groups: AMI, 27 UAP, 43 AP, 22 NIP and 18 excluded. Albumin and creatinine were measured in the first urine passed after admission (sample 1) and the first morning urine the following 2 days (samples 2 and 3). The ACR was significantly higher in the AMI and UAP groups than in the other patient groups (p < 0.0001). There was no significant difference of ACR between the AMI and UAP in sample 1 (p = 0.31). In the AMI, UAP and AP groups ACR was significantly higher in sample 1 than in samples 2 and 3 (p < 0.005). In the NIP group there were no significant differences between sample 1 versus samples 2 and 3 (p = 0.06). Series 2 comprised 10 patients: 8 AMI, 1 UAP and 1 AMYO. ACR were measured in all specimens voided during the period of observation. ACR can oscillate within hours between normal concentrations and concentrations well into or above the microalbuminuric range. We propose the term episodic albuminuria for this reversible, switch-like change in renal function. The albuminuric episodes lasted 90-600 minutes. Maximum values for ACR were between 133-790 mumol/mol or 78-466 mg/g. In healthy, resting individuals ACR is < 50 mumol/mol (< 30 mg/g). The rapid changes in glomerular permeability may reflect systemic changes in endothelial permeability in the affected individuals. We speculate that atrial natriuretic peptide (ANP) may be a mediator of this type of albuminuria. PMID- 10389214 TI - Genetic prediction of coronary heart disease: lessons from Canada. AB - Before it can find clinical application, a panel of genetic tests to predict coronary heart disease (CHD) risk will need to be shown to be more effective than either a determination of family history or the assessment of intermediate phenotypes, such as plasma lipoproteins, diabetes and blood pressure. Our studies in Canadian Hutterites and Oji-Cree indicate that a single genetic factor has a small effect on an intermediate phenotype of CHD. Also, the total contribution of genetic factors, while substantial, is usually the aggregate of many small effects. Furthermore, the environment plays a capacious role in modulating the expression of the genetic susceptibility to CHD. This is seen in examples of strong monogenic determinants of CHD, such as in early CHD in Newfoundlanders that results from APOA1 Q[-2]X, and in early onset diabetes in Oji-Cree that is determined by HNF-1 alpha S319. The Oji-Cree HNF-1 alpha S3 19 example also indicates that future diagnostic tests will have to account for population specific genetic determinants of CHD risk. Studies in Canadian Inuit suggest that a prudent environment can override an apparently high level of genetic susceptibility to CHD. These anecdotes imply that environment might be an even greater determinant of the common polygenic forms of CHD. The complexity of CHD will produce a delay in implementing routine genetic testing. This might provide more time for health care providers and society to consider the possible implications--medical, ethical and legal--and limitations of the genetic prediction of CHD. PMID- 10389215 TI - Multilocus approach to cardiovascular risk. AB - Until now, our familial studies have showed that shared genetic and environmental factors are involved on lipid parameters variability. More precisely, being working on 119 families we have showed that: a) The apolipoprotein E (apo E) common polymorphism is involved in the total cholesterol, low density lipoprotein cholesterol (LDL-Chol), apo E, apo B levels variability, b) the apolipoprotein A IV gene had no effect on lipid metabolism parameters variability, apo A-IV levels included, c) the apolipoprotein B gene was associated with total cholesterol, high density lipoprotein cholesterol, LDL-Chol, triglycerides and apo B levels genetic variability, d) the lipoproteine lipase (LPL) gene was responsible for 6.5% of the triglycerides variability, e) the apo E and LPL 447 polymorphisms influence in conjunction lipid parameters. These preliminary results on effects and combination effects of polymorphic genes show the interest of a multilocus approach. We have used in a subgroup of 416 individuals of a familial cohort (Stanislas Cohort) a prototype assay that genotypes a panel of 35 polymorphic sites on 15 candidate genes of Cardiovascular diseases. Each sample is amplified by two multiplex polymerase chain reactions, then hybridized to an array of immobilized, oligonucleotide probes. The frequencies of the rare alleles were in agreement with those reported by others in caucasian populations. The realisation of this multiplex assay in the 1,006 families of the Stanislas Cohort, which is underway, will allow us a better understanding of the inter-individual variability of lipids and will contribute to the determination of the genetic susceptibility of one's individual to cardiovascular risk. PMID- 10389216 TI - Development of a novel, N-terminal-proBNP (NT-proBNP) assay with a low detection limit. AB - A novel, highly sensitive and specific N-Terminal-proBNP (NT-proBNP) assay based on a sandwich format has been developed. The assay time is below 2 hours and no extraction process is needed. The calibration curve covers a NT-proBNP concentration range from 0 pmol/L up to 600 pmol/L. The analytical detection limit of the assay was estimated to be 2.7 pmol/L (3 SD). The intra-assay coefficient of variation is 5.7% (at 50 pmol/L) and 6.1% (at 250 pmol/L), while the inter-assay CVs are 15.8% (15 pmol/L) and 8.2% (250 pmol/L). There is no significant interference by bilirubin (up to 900 mumol/L), haemoglobin (up to 10 g/L), rheumatoid factors (up to 975 IU/mL), triglycerides (up to 20.5 mmol/L), biotin (up to 50 micrograms/L), digoxin (up to 100 micrograms/L) and digitoxin (up to 200 micrograms/L). The analyte NT-proBNP is fully stable in whole blood over 3 days and in EDTA-plasma over 24 hours. This good stability of NT-proBNP compared to other less stable natriuretic peptides is a significant advantage and a main prerequisite for a routine diagnostic marker. Preliminary results of using this new assay in clinical studies for diagnosing and monitoring left ventricular dysfunction demonstrate that there is a significant gain in diagnostic validity. PMID- 10389217 TI - The effects of spinal or mesencephalic transections on sleep-related erections and ex-copula penile reflexes in the rat. AB - The neural mechanisms of penile erections during paradoxical sleep (PS) remain unknown since it has yet to be the subject of neurophysiological investigation. Using a new experimental model for sleep-related erection research in freely behaving rats, neural transections were undertaken to definitively elucidate the effects of paraplegia on PS-related erections and to determine at which brain level the mechanisms underlying PS erectile activity are generated. Continuous polygraphic recordings, as well as ex-copula penile reflexes, were performed in male Sprague Dawley rats before and after spinal (n = 4) or mesencephalic (n = 6) transections. Spinal transections virtually eliminated PS-related erections. Following mesencephalic transections, PS remained qualitatively intact in all rats. PS erectile activity, however, was severely disrupted, as shown by a significant decrease in the total number of erections, the number of erections per hour, and the percentage of PS phases exhibiting an erectile event. Finally, spinal and mesencephalic transections had contrasting effects on ex-copula penile reflexes. Spinal transections significantly shortened the latency to reflex induction and increased the percentage of tests eliciting an erectile event, whereas mesencephalic transections significantly increased the latency to reflex induction without affecting the percentage of tests eliciting an erectile event. These data suggest that the brainstem is not sufficient for the generation of PS erectile activity even though it is sufficient for the generation of other classic PS phenomena. We conclude that neural structures rostral to the mesencephalopn (i.e., the forebrain) are essential for the maintenance and integrity of PS related-erections. The reflex erection data suggest that spinal transection removes a tonic descending inhibition of erections, whereas such an inhibition not only remains intact, but appears enhanced following mesencephalic transection. We hypothesize that the forebrain plays a facilitatory role in erectile control, at least in part, through disinhibition of brainstem tonic anti erectile mechanisms. PMID- 10389218 TI - Comparison of MK-801 and sleep deprivation effects on NREM, REM, and waking spectra in the rat. AB - In previous studies, we showed that blockade of the cation channel gated by NMDA glutamate receptors with ketamine or MK-801 massively stimulates NREM delta. We now test whether this NREM delta stimulation is physiological by comparing the EEG response following MK-801 to the EEG response following sleep deprivation (SD). Our previous studies measured only NREM 1-4 Hz EEG with period-amplitude analysis (PAA). Here we extended the analysis of MK-801 effects on sleep EEG by applying power spectral analysis (PSA) to examine delta and higher frequency spectra (.2-100 Hz) in NREM and by including REM and waking spectra. The changes in EEG spectra following MK-801 and SD were remarkably similar. Both SD and MK 801 produced their largest changes in NREM delta and REM 10-20 Hz power. There were some differences in the high frequency EEG, but the overall similarity of the PSA spectra in all three vigilance states after MK-801 and SD supports the possibility that MK-801 stimulated physiologic sleep, perhaps by increasing the need for homeostatic recovery from the metabolic effects of NMDA channel blockade. PMID- 10389219 TI - Post-apneic inhalation reverses apnea-induced sympathoexcitation before restoration of blood oxygen levels. AB - Sleep apneas acutely increase sympathetic nerve activity (SNA) and thus arterial blood pressure. We hypothesized that after apnea, sympathoexcitation decreases before recovery of blood oxygen levels because of the predominant inhibitory effect respiratory factors exert over sympathetic nervous system activation. Seven healthy subjects were instrumented for arterial oxygen saturation (pulse oximetry, SaO2), leg muscle SNA (microneurography), and arterial pressure (Finapres). Supine subjects breathed 12% oxygen, 3% carbon dioxide, and 85% nitrogen for one min prior to apnea at the end of a normal tidal expiration. We accounted for circulatory delay in SaO2 measurement (5.4 +/- 0.4 s, mean +/- SE) as the time from the termination of apnea to the midpoint of the nadir of SaO2. SaO2 decreased to average 84 +/- 3% over the final 10 seconds of apnea, and recovered only partially to average 87 +/- 3% over the 10 seconds immediately following apnea. End-expiratory apnea increased SNA 14-fold from baseline levels of 217 +/- 37 units/10 seconds to 3063 +/- 442 units/10 seconds. However, SNA decreased to 93 +/- 32 units/10 s during the first 10 seconds after apnea. These findings indicate that sympathoinhibitory effects of respiratory signals, either lung inflation receptors or central respiratory inputs, predominate over sympathoexcitatory inputs from chemoreceptors to produce immediate and complete sympathoinhibition at the termination of a voluntary apnea. Arterial baroreflexes probably also contribute to sympathoinhibition after apnea. PMID- 10389220 TI - Spectral indices of cardiac autonomic function in obstructive sleep apnea. AB - Spectral analysis of heart rate variability (HRV) is useful as a noninvasive means of assessing autonomic function in patients with obstructive sleep apnea (OSA). However, standard spectral measures, such as the ratio of low-frequency to high-frequency power (LHR) and normalized high-frequency power (NHFP), can be confounded by the abnormal breathing patterns that occur during sleep. To circumvent this limitation, we employed an autoregressive modeling approach to partition the RR time-series into a component that is correlated with respiration and a respiration-independent component. From these components, we derived two new spectral indices: the modified LHR (MLHR) and the average gain relating respiration to RR changes (GRSA). Six normals and seven OSA patients were studied in relaxed wakefulness and stage 2 sleep; during sleep, the OSA patients were studied without and with continuous positive airway pressure (CPAP) therapy. All four spectral indices showed significant differences between OSA patients and normals in both wakefulness and sleep, although the changes in MLHR and GRSA were substantially larger and less variable: MLHR (p < 0.0003) and GRSA (p < 0.0001) vs. LHR (p < 0.005) and NHFP (p < 0.004). However, in the OSA subjects, LHR and NHFP were unchanged by CPAP. By contrast, CPAP produced a highly significant increase in GRSA (p < 0.0004), as well as a decrease in MLHR (p < 0.03). Thus, by compensating for the effects of breathing pattern differences, MLHR and GRSA unmasked the effects of CPAP therapy, which has been shown in previous studies to reduce sympathetic activity and increase vagal cardiac modulation. PMID- 10389221 TI - Disruption of normal gastric myoelectric functioning by sleep. AB - STUDY OBJECTIVES: The aim of this study was to assess the effects of sleep on gastric myoelectric activity as measured by electrogastrography in healthy individuals. The goal was to elucidate the role of central influences in the regulation of normal gastric functioning. DESIGN: Electrograstrogram (EGG) was recorded during polysomnographically monitored waking and sleep. SETTING: Sleep laboratory. PARTICIPANTS: 17 healthy volunteers. MEASUREMENTS AND RESULTS: EGG parameters were computed for 20-minute segments of pre-sleep waking, stage 2 sleep, stage 4 sleep, and REM sleep using both overall and running spectral analysis of EGG data. The dominant power decreased significantly from waking (31.4 +/- 1.4 dB) to all sleep stages (23.1 +/- 1.5 dB during stage 2; 24.7 +/- 1.4 dB during stage 4; 24.3 +/- 1.3 dB during REM sleep). The percentage of 2 4cpm activity decreased significantly during NREM sleep (64.6 +/- 7.6% during stage 2 sleep; 57.5 +/- 5.5% during stage 4 sleep) compared to its waking value (90.8 +/- 3.2%), but not compared to REM sleep (74.1 +/- 5.4%). The instability coefficient of the dominant frequency increased significantly from waking (0.19 +/- 0.03) to all sleep stages (0.36 +/- 0.05 during stage 2 sleep; 0.47 +/- 0.05 during stage 4; 0.34 +/- 0.05 during REM sleep). No significant differences between the sleep stages were found for any measure. CONCLUSIONS: Sleep is associated with increases in gastric dysrhythmia and instability of the gastric slow wave frequency when compared to waking. These findings suggest that the intrinsic electrical activity of the stomach is significantly influenced by central nervous system mechanisms, and support the notion of a brain-gut axis. PMID- 10389222 TI - Spontaneous baroreflex analysis in non-apneic snoring individuals during NREM sleep. AB - The primary purpose of this study was to measure baroreceptor sensitivity (BS) during wakefulness and non-rapid eye movement (NREM) sleep in non-apneic snoring individuals. To achieve this purpose continuous and simultaneous measurements of snoring, oxygen saturation, sleep stages, arterial blood pressure and heart rate were obtained from seven non-apneic snoring subjects. After obtaining these measures, a computer program was employed to detect concomitant increases or decreases in systolic blood pressure and R-R interval duration during sequences of three or more consecutive beats that occurred during stage II and slow wave sleep (SWS). The values recorded from a given sequence were plotted and the slope of the regression line fit to the data was used as a measure of BS. The results showed that mean arterial pressure and heart rate during stage II and SWS of NREM sleep were not significantly different from wakefulness. In contrast, the BS measured during NREM sleep was significantly lower than values recorded during wakefulness. In addition, linear regression analysis showed that an inverse and significant correlation existed between snoring frequency and the decrease in BS during sleep. We conclude that the decrease in blood pressure and heart rate normally observed during NREM sleep in healthy non-snoring individuals is attenuated or abolished in non-apneic snoring individuals and that these cardiovascular alterations may be partially mediated by a decrease in BS. PMID- 10389224 TI - Epidemiology of medication as aids to alertness in early adulthood. PMID- 10389223 TI - Long distance driving and self-induced sleep deprivation among automobile drivers. AB - OBJECTIVE: To evaluate the sleep hygiene and prevalence of sleep deprivation among a large sample of automobile drivers. DESIGN: From the 15th of June to the 4th of August 1996, with the help of the French highway patrol, we randomly stopped automobile drivers at the toll booths of Bordeaux and Biarritz. All subjects completed a validated questionnaire on sleep/wake habits during the year. After answering the questionnaire, subjects completed a graphic travel and sleep log of the three days preceding the interview. PARTICIPANTS: We randomly stopped 2196 automobile drivers. Ninety-one percent of the sample (mean age 43 +/ 13 years) agreed to participate in the survey. RESULTS: Fifty percent of the drivers decreased their total sleep time in the 24 hours before the interview compared with their regular self-reported sleep time. 12.5% presented a sleep debt > 180 minutes, and 2.7% presented a sleep debt > 300 minutes. Being young, commuting to work, driving long distances, starting the trip at night, being an "evening" person, being a long sleeper during the week, and sleeping in on the week-end were risk factors significantly associated with sleep debt. CONCLUSION: The results of the study highlight variables (long-distance driving, youth, sleep restriction) that are frequently associated with sleep-related accidents. PMID- 10389225 TI - Age, working conditions, and sleep disorders: a longitudinal analysis in the French cohort E.S.T.E.V. AB - STUDY OBJECTIVES: To investigate the effects of occupational factors on both the incidence and the disappearance of sleep disorders after a five-year follow-up period. DESIGN: A prospective longitudinal investigation E.S.T.E.V. carried out in 1990 and 1995. SETTING: Seven regions of France. SUBJECTS: A random sample of employed men and women born in 1938, 1943, 1948, and 1953. In 1990, 21,378 subjects were interviewed (87% of those contacted), and 88% were interviewed again in 1995. MEASURES: Sleep disorders (SD), objectifiable and psychosocial working conditions. RESULTS: Prevalence of SD increased with age and were more frequent among women than men in every age group. Incidence of SD varied little with age, but their disappearance decreased with age. After adjustment for age and sex, SD in 1995 were found to be associated both with objectifiable working conditions and with psychosocial aspects of the way work is experienced. Among objectifiable occupational risk factors, shift work, work week often longer than 48 hours, and exposure to vibrations appeared to be the principal risk factors for SD. Among psychosocial occupational factors, finding it difficult or irksome to have to hurry appeared to be the principal risk factor. CONCLUSIONS: Taking into account the adjustments for health criteria, sociodemographic characteristics, and leisure activities, these results suggest useful courses of action for prevention which, it seems to us, must not be only limited to objectifiable working conditions. Issues about work organization, while clearly difficult to resolve, must also be taken into account. PMID- 10389226 TI - Effect of celiprolol treatment in hypertensive patients with sleep apnea. AB - The effects of a beta-blocker, celiprolol, on sleep and arterial blood pressure (BP) were evaluated during a single-blind study in seven hypertensive patients with sleep apnea. Diurnal ambulatory BP measurements with an automatic cuff inflation device and polysomnography with simultaneous Finapres BP recording were performed separately on consecutive days at the end of two 21-day treatment periods involving placebo followed by celiprolol (200 mg/day). Age was 59 +/- 2.5 yr (m +/- sem) and body mass index 33.2 +/- 2.3 kg. m-2. Diurnal ambulatory BP was significantly lower with celiprolol than with placebo (systolic 139 +/- 4 vs 152 +/- 5 mmHg, diastolic 86 +/- 2 vs 96 +/- 2 mmHg). The apnea-hypopnea index was similar under celiprolol and placebo (48 +/- 7.4 vs 53 +/- 7.8, respectively), as were the total sleep time and percent of duration of the different sleep stages. Individual average BP values were significantly lower during REM sleep under celiprolol but remained similar under celiprolol and placebo in the other sleep stages. Variability of nocturnal BP (assessed by the SD of distribution of BP variations) was not affected by celiprolol. In conclusion, celiprolol which decreased daytime BP, did not affect sleep pattern or respiratory disturbances, or nocturnal BP variability related to apnea. PMID- 10389227 TI - Transdermal scopolamine alters phasic REM activity in normal young adults. AB - Transdermal scopolamine has been widely used for the prevention of motion sickness by travelers due to its potent anticholinergic effects and the ease of administration. Nevertheless, its effects on sleep physiology are not known, despite the wellknown fact that the administration of scopolamine as an injection or an oral form could influence the sleep architecture, especially prolonging rapid eye movement (REM) sleep latency and shortening duration of REM sleep. This study aimed to measure the influence of transdermal scopolamine on REM sleep in order to examine whether it affects REM sleep as in the previous studies. We studied eight young healthy male adults polysomnographically for three nights including one adaptation night in a double blind crossover design and compared REM sleep-related variables between sleeps with and without scopolamine patch of 1.5 mg. We found no differences in tonic REM sleep measures such as REM duration and REM latency, but phasic REM sleep measures such as total REM activity (p < 0.05) and total REM density (p < 0.05) were significantly suppressed by the transdermal scopolamine; REM densities of the first (p < 0.05) and the second (p < 0.05) REM sleep periods as well as REM activity of the fourth REM sleep period (p < 0.05) were decreased significantly on the scopolamine patch nights compared with placebo patch nights. These results suggest that phasic REM components reflect cholinergic mechanism in the central nervous system (CNS) even at the lowest commercial dose, and could be useful markers of CNS cholinergic activities in the future research. PMID- 10389228 TI - T cell receptors in health and disease. Introduction. PMID- 10389229 TI - T cell receptor usage in autoimmune disease. PMID- 10389230 TI - T cell receptor usage in malignant diseases. PMID- 10389231 TI - T cell receptor usage in infectious disease. PMID- 10389232 TI - gamma delta T cells, their T cell receptor usage and role in human diseases. PMID- 10389233 TI - Treatments targeting the T cell receptor (TCR): effects of TCR peptide-specific T cells on activation, migration, and encephalitogenicity of myelin basic protein specific T cells. PMID- 10389234 TI - On the reporting of dental health, time for dental care, and the treatment panorama. AB - The thesis included five methodological studies and one caries epidemiological investigation, the general aim being to study how to measure and report dental health, time for dental care, treatment panorama, and dental care outcomes, within a Public Dental Service organization. The specific aims were to monitor dental clinic activities using a time study method, to apply time study results of a dental health-related patient group system for the 3-19 year age groups, and to compare time study results with corresponding results from computerized systems used for reporting dental care. Other specific aims were to compare longitudinal caries index data results between cohort and cross-sectional samples, to analyse caries index for extreme caries groups among adolescents leaving organized dental care, and--using time series methods--to analyse dental health development of the 15-19 year age groups. Results from the time studies portrayed the dental clinic as a working unit, showed that reported values can represent dental care only for intervention procedures, and indicated that clinic patterns were not adapted to the health situation of the patient groups. Longitudinal cohort attempts gave different values from those of the cross sectional year classes, which should be the primary focus when presenting caries index mean values in dental health reviews. Caries-free groups from 15 to 19 years of age seem to be stable in their caries development in about 60%-80% of cases; while the 20% groups with the highest index values accounted for about 80% of all approximal lesions. In times of major economic adjustment, dental health for adolescents in Goteborg was an example of sustainable dental health development. A model system for monitoring, analysing, and reporting dental health and dental care outcomes within a dental care-giving organization calls for several conditions, for example, a dental health-related patient group system, and a rationale for the choice of dental team models. These areas could be gathered into a system where contemporary socio-economic factors and dental research results interact with performed dental care, and also with different methods for reporting and evaluating dental health, dental care costs, and the demand for dental care competence. PMID- 10389235 TI - Autocatalytic replication in a CSTR and constant organization. AB - The dynamics of a network of autocatalytically replicating species in a continuously stirred tank reactor can be described by a replicator equation in the limit of small flux rates. PMID- 10389236 TI - Mathematical analysis of binary activation of a cell cycle kinase which down regulates its own inhibitor. AB - In mammalian cells, the heterodimeric kinase cyclin E/CDK2 (EK2) mediates cell cycle progress from G1 phase into S phase. The protein p27Kip1 (p27) binds to and inhibits EK2; but EK2 can phosphorylate p27, and that leads to the deactivation of p27, presumably liberating more EK2 and forming a positive-feedback loop. It has been proposed that this positive-feedback loop gives rise to binary (all-or none) release of EK2 from its inactive complex with p27. Binary release suggests a bistable biochemical system in which a stable steady state with low EK2 activity is extinguished in a saddle-node bifurcation, causing the system to shift abruptly to a stable steady state with high EK2 activity. Two mathematical models are discussed, one in which free EK2 deactivates p27 in the EK2-p27 inhibitory complex as well as free p27, and one in which the rate of EK2 catalyzed deactivation of free p27 has saturable kinetics with respect to free p27. In general, if inhibitory binding is approximately in equilibrium, bistability requires that there be a potential unstable steady state where the reaction order of p27 deactivation is greater with respect to EK2 than with respect to p27. PMID- 10389237 TI - Partitioning of four modern volatile general anesthetics into solvents that model buried amino acid side-chains. AB - Partitioning of four modern inhalational anesthetics (halothane, isoflurane, enflurane, and sevoflurane) between the gas phase and nine organic solvents that model different amino acid side-chains and lipid membrane domains was performed in an effort to define which microenvironments present in proteins and lipid bilayers might be favored. Compared to a purely aliphatic environment (hexane), the presence of an aromatic-, alcohol-, thiol- or sulfide group on the solvent improved anesthetic partitioning, by factors of 1.3-5.2 for halothane, 1.7-5.6 for isoflurane, 1.7-7.6 for enflurane, and 1.5-7.3 for sevoflurane. The most favorable solvent for halothane partitioning was ethyl methyl sulfide, a model for methionine. Enflurane and isoflurane partitioned most extensively into methanol, a model for serine, and sevoflurane into ethanol, a model for threonine. Isoflurane also partitioned favorably into ethyl methyl sulfide. The results suggest that volatile general anesthetics interact better with partly polar groups, which are present on amino acids frequently found buried in the hydrophobic core of proteins, compared to purely aliphatic side-chains. Furthermore, if an anesthetic molecule was located in a saturated region of a phospholipid bilayer membrane, there would be an energetically favorable driving force for it to move into several higher dielectric microenvironments present on membrane proteins. The results provide evidence that proteins rather than lipids are the likely targets of volatile general anesthetics in biological membranes. PMID- 10389238 TI - Effective diffusion coefficients of K+ and Cl- ions in ion channel models. AB - We have used molecular dynamics simulations, corresponding to a total simulation time of 11 ns, to investigate the effective short-time local diffusion coefficient of potassium and chloride ions in a series of model ion channels. These models, which include channels formed by the fungal peptide alamethicin, by a synthetic leucine-serine peptide, and by the pore-lining M2 helix bundle of the nicotinic acetylcholine receptor, have a range of different secondary structures, diameters and hydrophobicities. We find that the diffusion coefficients of both ions are appreciably reduced in the narrower channels, the extent of the reduction being similar for both the anionic and cationic species. This suggests that a difference in mobility cannot be the source of the ion selectivity exhibited by some of the channels (for example, the leucine-serine peptide). We find no evidence for a reduction in mobility of either ion in the nAChR model. These results are broadly in line with a previous similar study of Na+ ions, and may be useful in Poisson-Nernst-Planck, Eyring rate theory or Brownian dynamics calculations of channel conductance. PMID- 10389240 TI - Semi-quantitative characterization of electroporation-assisted disinfection processes for inactivation of Giardia and Cryptosporidium. AB - The effect of electroporation (very short duration pulses of high voltage electricity) on the viability of Giardia cysts and Cryptosporidium oocysts, and on the viability of these organisms in the presence of free chlorine, combined chlorine, hydrogen peroxide and potassium permanganate, was examined. While electroporation itself had only a minor effect on survival, the combination of electrical and chemical treatment produced superior inactivation, particularly with combined chlorine, hydrogen peroxide and potassium permanganate. This enhancement may provide a relatively practical way of achieving enhanced inactivation of resistant protozoa by water disinfection processes. Further study of kinetics and optimum treatment combinations is needed. PMID- 10389239 TI - Membrane destabilizing activity of influenza virus hemagglutinin-based synthetic peptide: implications of critical glycine residue in fusion peptide. AB - Peptide III is a 20-residue synthetic model peptide based on the fusion peptide of influenza virus A/PR/8/34 strain and takes a secondary structure similar to the original peptide. While conserving the amphiphilic helical nature, 20 peptides to modify the bulkiness of side chains of peptide III were synthesized, and acid-induced membrane destabilization was assessed by aqueous content leakage from large unilamellar vesicles. Substitutions on the hydrophobic side decreased activity but showed less effect on the hydrophilic side, which confirmed the importance of the hydrophobic side for interaction with the membrane. Interestingly, substitution at the 13th Gly residue enhanced the amphiphilic helical nature but severely reduced activity. Correlation between alpha-helical content at acidic pH and the activity was not recognized, suggesting rather that the importance of this site was due to helix termination by glycine which allows N-terminal and C-terminal halves to behave as different secondary structural units. PMID- 10389241 TI - Viable heterotrophic bacteria in water and sediment in 'Mar Piccolo' of Taranto (Ionian Sea, Italy). AB - Samples of water and sediment were collected from October 1996 to September 1997 in 'Mar Piccolo' of Taranto (Ionian Sea, Italy). Mar Piccolo is a semi-enclosed basin subject to pollution and receives a considerable amount of sewage and industrial waste. Qualitative and quantitative analyses of the composition of the microbial flora were conducted on samples from six stations. The highest bacterial densities, in water and in sediment samples, were found in summer and the lowest, in autumn. Among Gram-negative bacteria, the predominant genus was Aeromonas; Photobacterium and Pseudomonas were also found. Gram-positive bacilli were abundant at all sampling points. Faecal contamination indicators demonstrated that all the stations examined in Mar Piccolo are influenced by anthropogenic pollution throughout the year. PMID- 10389242 TI - The role of surface physicochemical properties in determining the distribution of the autochthonous microflora in mineral water bottles. AB - Investigation of the distribution of the viable autochthonous microflora in three brands of 1-2-month-old bottled mineral water showed that 1.8 x 10(4) (S.E.M. 8.9 x 10(3), n = 5) to 1.2 x 10(5) (S.E.M. 1.3 x 10(4), n = 5) cfu ml-1 were planktonic cells while 11 (S.E.M. 4, n = 5)-632 (S.E.M. 176, n = 5) cfu cm-2 were found in the biofilm. The biofilm represented between 0.03 and 1.79% of the total viable microbial population in the 1.5 litre bottles studied. Scanning electron microscopy studies showed that the cells adhering to the polyethylene terephthalate (PET) bottles were predominantly rod-shaped, sparsely distributed over the surface. In contrast, the cells adhering to the high density polyethylene (HDPE) caps were found to be mainly clumps of coccoid cells, suggesting that the bottle may provide different microhabitats for different microfloras. Large-scale roughness, such as that observed as lettering inside the cap (average height (z) = 93 microns) was associated with a 46-fold increase in cell numbers. Increased small-scale roughness, as measured by atomic force microscopy on PET and HDPE surfaces (average roughness (Ra) = 5-551 (nm), showed no correlation with adhesion. Investigations of surface hydrophobicity by the sessile drop technique showed that contact angles (theta) were greater on the HDPE caps (theta = 89-96 degrees) than on the PET surfaces (theta = 69-80 degrees). However, no correlation was found between contact angle and attached cell numbers. Measurements of surface electrostatic charge by streaming potential showed that the PET carried an overall negative charge, measuring -15.9 to -16.6 mV in mineral water. No significant change in charge occurred when the monomer composition of the PET was altered. It was concluded that surface roughness, in particular the scale of surface topographical features, is the most important physicochemical surface characteristic determining the distribution of the autochthonous microflora in mineral water bottles. PMID- 10389243 TI - Effect of nutrient limitation on adhesion characteristics of Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa causes a variety of diseases in humans including lung and ocular infections. Infections of the cornea are usually associated with wearing contact lenses and can result in loss of vision. This study aimed to determine the effect of carbon or nitrogen limitation on the adhesion to contact lenses of a strain of Ps. aeruginosa isolated from contact lens-related corneal inflammation. Cells were grown in a continuous culture apparatus in varying levels of glucose or ammonia to effect nutrient limitation. Adhesion to contact lenses was measured as total counts and viable counts. The cell surface hydrophobicity and charge were measured using adhesion to surface-modified Sepharose. Changes in lipopolysaccharide were determined using 1D SDS-PAGE and changes in cell-surface proteins were measured using 2D gel electrophoresis. The more the cultures were nitrogen limited, the greater the increase in adhesion to unworn hydrogel contact lenses 0.3 x 10(3) - 2.2 x 10(3) cells/mm2 on Etafilon A lenses. Cells that were carbon limited showed a greater increase in adhesion to contact lenses when the lenses had been coated in artificial tears. It appeared that lipopolysaccharide may have been involved in the constitutive adhesion to unworn lenses that occurred during C-limitation, whereas changes in the outer membrane proteins contributed to the increased adhesion under nitrogen limitation, or the change in adhesion that occurred to carbon-limited cells using contact lenses coated in artificial tears. Nine cell-surface proteins appeared during nitrogen limitation with kDa/pI of 75/4.8, 4.9, 5.0; 62/5.6; 89/6.5; 38/6.4; 28/1.5; 18/6.4; 12/4.5. Any or all of these may have been involved in the increased adhesion and further experiments are underway to examine this possibility. PMID- 10389244 TI - Evaluation of a portable differential continuous flow centrifuge for concentration of Cryptosporidium oocysts and Giardia cysts from water. AB - A portable device was developed and assembled from a stationary differential continuous flow centrifuge usually employed for blood cell separation, for the purpose of concentrating Cryptosporidium and Giardia from large volumes of water. Following compaction onto the wall of the disposable plastic centrifuge bowl and aspiration of residual water, the oocysts and cysts were dislodged by injection of a 20 ml solution containing 0.01% Tween-80 and 1% SDS and vigorous shaking. Following aspiration, the oocysts were pelleted, reacted with specific FITC conjugated monoclonal antibodies, and enumerated via fluorescence microscopy. The entire procedure required about 2 h. Initially, 55% and 87% of Cryptosporidium oocysts and Giardia cysts, respectively, were recovered from 45 litres of tap water, and 27% and 57%, respectively, from river water. Adjustments in centrifuge speed and flow rates improved recovery to about 90% for Cryptosporidium oocysts and hence, this method compared favourably with the recently developed calcium carbonate flocculation method. It was superior in time requirement and volume flexibility, and showed a distinct advantage over the standard cartridge filtration method in all respects. The continuous flow centrifugation equipment is compact, mobile, flexible, and yields reproducibly high recovery rates. The ease of handling, speed of performance and minimal requirements for post concentration equipment, reagents and labour make the system highly cost effective. It appears to offer an improved method, well suited for use by water utilities for monitoring the burden of water-borne protozoan pathogens. PMID- 10389245 TI - Performance of open-fronted microbiological safety cabinets: the value of operator protection tests during routine servicing. AB - The performance of class I and II microbiological safety cabinets over 7 years, employed in a force-ventilated containment level 3 (CL-3) laboratory, is described. Operator Protection (OP) provided by the cabinets, assessed by still and latterly limited 'in-use' KI-Discus tests, showed no overall deterioration during the review period. Comparisons show that a selected class II unit, but not a second, and a new class II MSC in a recently commissioned, similar CL-3 facility, provide the same order of OP as a class I cabinet. From the experiences described, it is strongly recommended that OP tests (OPTs) should be part of the routine servicing regime to ensure that cabinets meet required performance levels, and additionally to allow detection and rectification of poor containment, particularly where induced by environmental factors. The value of OPTs is discussed with reference to certain national standards. PMID- 10389246 TI - NO3- nutrition and salt tolerance in the cyanobacterium Anabaena sp. PCC 7120 and mutant strains. AB - Growth of the cyanobacterium Anabaena sp. PCC 7120 and its nitrate assimilation defective mutants was inversely proportional to the NaCl concentration in the medium. Presence of nitrate in the saline medium protected the growth of the parent but not of the mutant strains from salt toxicity. On the other hand, ammonium nitrogen protected the growth of all the strains from salt toxicity. However, the effect was less than that of nitrate. An altered sodium transport system was evident in the mutant strains and was most marked in mutant SP9. The cellular sodium concentration in parent and mutant strains also varied. Although mutant SP9 exhibited the lowest level of cellular sodium, it was as sensitive to salt toxicity as other strains. It is assumed that merely the presence of a toxic level of NaCl in the ambient environment is sufficient to damage the structural and functional components of the plasma membrane. PMID- 10389247 TI - Low level chemiluminescence from liquid culture media. AB - Low level chemiluminescence (CL) can be observed from autoclaved liquid culture media, as used in microbiology. The light emission is oxygen-dependent and arises from reactions following auto-oxidation of reducing Maillard products which are formed during autoclaving. The inhibition of this CL by radical scavengers and antioxidants has been studied. As superoxide radicals and hydrogen peroxide are predominantly involved in the initiation of the CL, the investigation of CL from culture media offers a convenient tool for the detection of exogenous (medium mediated) oxidative stress being imposed onto micro-organisms in culture. Transition metal ions showed, dependent on concentration, both inhibitory and stimulating effects on the CL, which was also affected by the presence of complexing agents. Iron porphyrins and related complexes displayed a very efficient quenching of the CL, which may be of particular importance, as aerobic micro-organisms have been previously shown to be very efficient in quenching the CL from culture media. PMID- 10389248 TI - Biodegradation of gasoline: kinetics, mass balance and fate of individual hydrocarbons. AB - The degradation of gasoline by a microflora from an urban waste water activated sludge was investigated in detail. Degradation kinetics were studied in liquid cultures at 30 degrees C by determination of overall O2 consumption and CO2 production and by chromatographic analysis of all 83 identifiable compounds. In a first fast phase (2 d) of biodegradation, 74% of gasoline, involving mostly aromatic hydrocarbons, was consumed. A further 20%, involving other hydrocarbons, was consumed in a second slow phase (23 d). Undegraded compounds (6% of gasoline) were essentially some branched alkanes with a quaternary carbon or/and alkyl chains on consecutive carbons but cycloalkanes, alkenes and C10- and C11 alkylated benzenes were degraded. The degradation kinetics of individual hydrocarbons, determined in separate incubations, followed patterns similar to those observed in cultures on gasoline. Carbon balance experiments of gasoline degradation were performed. The carbon of degraded gasoline was mainly (61.7%) mineralized into CO2, the remaining carbon being essentially converted into biomass. PMID- 10389249 TI - Evaluation of the effect of temperature and nutrients on the survival of Campylobacter spp. in water microcosms. AB - Batch microcosms containing various water types (de-ionized and river water with or without sediment), incubated at a range of temperatures (5-37 degrees C), were used to facilitate a comparative evaluation of the significance of such variables and their interactions upon the collective and individual survival of four species of thermophilic Campylobacter. All variables significantly influenced (P < = 0.031) population decay rates. Minimal decay for the group was identified at low temperatures (5 degrees C) in river water, i.e. nutrient-containing microcosms. Collective decay rates within river water microcosms were significantly decreased (P = 0.03) from those observed in de-ionized water, particularly at environmental temperatures (5 and 15 degrees C). However, the increased nutrient levels observed in sediment-containing microcosms did not significantly (P = 0.41) reduce population decay rates. Overall, Camp. jejuni populations demonstrated the most resilience to the environmental stressors evaluated, with the exception of 15 degrees C where Camp. lari was the most persistent. Campylobacter coli and Camp. upsaliensis demonstrated comparable survival characteristics but were less resilient than Camp. jejuni and Camp. lari. These observations identify the suitability of water systems as a reservoir and medium for Campylobacter infection, and potentially identifies Camp. jejuni and Camp. lari as the main protagonists of water-mediated campylobacteriosis. PMID- 10389250 TI - Total counts, culturable and viable, and non-culturable microflora of a French mineral water: a case study. AB - The changes in bacterial counts during the storage of a natural mineral water from a French spring were studied. Samples were taken from the spring and the bottling line. Viable cultivable (VC) bacteria were counted on R2A medium. Total counts, viable and dead bacteria were counted using the LIVE/DEAD Bac Light VIABILITY kit and epifluorescence microscopy. Viable but non-cultivable (VNC) bacteria were estimated by difference between viable and VC counts. Isolates were clustered by phenotype. The microflora in the spring water increased from < 10-3 x 10(5) bacteria ml-1 after 6 d in storage and then stabilized. Mechanical bottling increased the allochthonous bacteria in the water that stabilized at 10(5) bacteria ml-1. Maximal growth is controlled by the low concentration of nutrients in the mineral water and the lysis of dead cells. The allochthonous bacteria came from the aquifer and colonized the filling line. The changes in the VC and VNC populations showed that the bacteria used starvation-survival and entry into the VNC state to adapt to the bottling stress and the enclosed oligotrophic environment. PMID- 10389251 TI - Ortho-phthalaldehyde: a possible alternative to glutaraldehyde for high level disinfection. AB - Ortho-phthalaldehyde (OPA) was tested against a range of organisms including glutaraldehyde-resistant mycobacteria, Bacillus subtilis spores and coat defective spores. Glutaraldehyde (GTA) and peracetic acid (PAA) were tested for comparative purposes. Both suspension and carrier tests were performed using a range of concentrations and exposure times. All three biocides were very effective (> or = 5 log reduction) against Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa in suspension tests. OPA and GTA (PAA was not tested) were also very effective against Staph. aureus and Ps. aeruginosa in carrier tests. OPA showed good activity against the mycobacteria tested including the two GTA-resistant strains, but 0.5% w/v OPA was found not to be sporicidal. However, limited activity was found with higher concentrations and pH values. Coat-defective spores were more susceptible to OPA, suggesting that the coat may be responsible for this resistance. The findings of this study suggest that OPA is effective against GTA-resistant mycobacteria and that it is a viable alternative to GTA for high level disinfection. PMID- 10389252 TI - Relationship between lipid composition, frequency of ethanol-induced respiratory deficient mutants, and ethanol tolerance in Saccharomyces cerevisiae. AB - The frequency of ethanol-induced respiratory deficient mutants and lipid composition in two Saccharomyces cerevisiae strains showing different degrees of ethanol tolerance were investigated. The more ethanol-tolerant strain exhibited a lower frequency of ethanol-induced respiratory deficient mutants than the less ethanol-tolerant strain. In addition, the more ethanol-tolerant strain contained a higher ergosterol/phospholipid ratio, a higher proportion of phosphatidylcholine, a lower proportion of phosphatidylethanolamine, a higher incorporation of long-chain fatty acids in total phospholipids, and a slightly higher proportion of unsaturated fatty acids in total phospholipids than the less ethanol-tolerant strain. These results show a clear relationship between the lipid composition, the frequency of ethanol-induced respiratory deficient mutants, and the ethanol tolerance of S. cerevisiae. A possible explanation of this relationship is discussed. PMID- 10389253 TI - Purification and partial amino acid sequence of plantaricin 1.25 alpha and 1.25 beta, two bacteriocins produced by Lactobacillus plantarum TMW1.25. AB - Two bacteriocins produced by Lactobacillus plantarum TMW1.25 have been purified by a four-step purification procedure, including ammonium sulphate precipitation and cation-exchange chromatography followed by hydrophobic-interaction chromatography on octyl sepharose. The final purification was performed by repeated reversed-phase chromatography steps which yielded two bacteriocin fractions designated plantaricin 1.25 alpha and plantaricin 1.25 beta. The molecular masses of the peptides in these fractions were 5979 and 5203 Da, respectively. Combination of the fractions did not have any synergistic effects on bacteriocin activity, indicating that they each contain a one-peptide bacteriocin. The major peptide in the alpha fraction was blocked at its N terminus, and a partial sequence (25 residues) could only be obtained after cleavage with CNBr. This sequence did not show clear homologies with known bacteriocins. The beta peptide has been sequenced almost completely and consists, presumably, of 53 residues. This peptide displayed strong homology to the known N terminal part of brevicin 27 produced by Lactobacillus brevis SB27. The results showed that the beta peptide contains as many as six consecutive lysine residues at the N-terminus. PMID- 10389254 TI - Evaluation of an alternative extraction procedure for enterotoxin determination in dairy products. AB - A concentration protocol based on trichloroacetic acid precipitation was evaluated and compared with the reference method using dialysis concentration. Different quantities of purified staphylococcal enterotoxins were added to pasteurized Camembert-type cheeses. Detection of enterotoxins in these cheeses was performed using an automated detection system. Raw goat milk Camembert-type cheeses involved in a staphylococcal food poisoning were also tested. Both enterotoxin extraction methods allowed detection of the lowest enterotoxin concentration level used in this study (0.5 ng g-1). Compared with the dialysis concentration method, TCA precipitation of staphylococcal enterotoxins was 'user friendly' and less time-consuming. These results suggest that TCA precipitation is a rapid (1 h), simple and reliable method of extracting enterotoxin from food which gives excellent recovery from dairy products. PMID- 10389255 TI - Detection of IgG antibody to bovine leukaemia virus in urine and serum by two enzyme immunoassays. AB - Four hundred blood sera from a cattle production unit were tested for BLV-(Bovine Leukaemia Virus) antibody with IP (Institut Porquier) and SB (Svanova Biotech) ELISA kits. Seventy-seven cattle with BLV-antibody (19.25%) and 77 without the antibody were used. No significant difference was found between O.D. of sera of PL+ (Persistent Lymphocytosis Positive) and PL- (Negative) cattle. The mean O.D. of urine samples of 77 seropositive cattle was significantly higher than that of 77 seronegative cattle (P < 0.01). There were also differences between urine O.D.s of seropositive (PL+) and seropositive (PL-) groups of cattle with IP (P < 0.05) and SB (P < 0.01) kits. All the results revealed the presence of BLV antibody in the urine of the cattle without any urinary dysfunction. PMID- 10389256 TI - Secretion of lysine in a broth medium by lactic bacteria and yeasts associated with garri production using a synthetic gene. AB - Although cassava is an important food in the tropics, it has two major deficiencies which are carried over into those foods made from it: its content of toxic cyanogenic glucosides and its low content of protein and amino acids. Garri, a fermented cassava food, has previously been ameliorated using organisms which simultaneously secrete linamarase (to reduce the residual cyanide in the food), amylase (to contribute to the growth of fermenting organisms and increase the flavour) and lysine (to improve the amino-acid content of the food). Some of the organisms fermenting cassava for garri production produce appreciable quantities of linamarase and amylase, but they are low in lysine production. It was therefore decided to improve these organisms by transforming them with a synthetic lysine gene coding for an 8-lysine peptide cloned in pBluescript II SK phagemid vector under the control of lac promoter. The synthetic lysine polypeptide gene was successfully introduced into Escherichia coli DH5 alpha and several strains of Lactobacillus spp. and Saccharomyces spp. There was a dramatic increase in lysine secretion in the organisms, ranging from about 2.5 to sixfold, following transformation with the synthetic gene. PMID- 10389258 TI - Cysteine protease is a major exotoxin of pathogenic luminous Vibrio harveyi in the tiger prawn, Penaeus monodon. AB - The role of an extracellular cysteine protease, produced by pathogenic luminous Vibrio harveyi strain 820514 originally isolated from diseased tiger prawn (Penaeus monodon), in the disease process in the prawns was studied. The protease was lethal to P. monodon with an LD50 value of 0.3 microgram protein g-1 prawn. The lethal toxicity of the extracellular products (ECP) of the bacterium was neutralized by pre-incubation of the ECP with rabbit antiserum to the cysteine protease. Pre-incubation of ECP with CuCl2 (an inhibitor of cysteine protease) also inhibited toxicity. This suggests that cysteine protease is the major toxin produced by the bacterium. The present protease is the first toxic cysteine protease to be found in Vibrio species. PMID- 10389257 TI - Inhibitory activity of Paenibacillus polymyxa SCE2 against human pathogenic micro organisms. AB - Paenibacillus polymyxa strain SCE2 was shown to inhibit the growth of different potential human pathogenic bacterial strains and fungi in vitro. To determine the genetic characterization of this antimicrobial substance, strain SCE2 was transformed with plasmid pTV32(Ts), a delivery vector for Tn917-lac. After transposition, four mutants were shown to have lost their capability to inhibit Micrococcus sp. and Staphylococcus aureus RN450, but they continued to inhibit the growth of Corynebacterium fimi NCTC7547 and Escherichia coli HB101. Hybridization experiments using the DNA of the four mutants digested with different endonucleases and pTV32(Ts) as a probe showed that the place of insertion of Tn917-lac in the chromosome was the same in mutants 4 and 36 and in mutants 31 and 59, but different between these pairs. It is thought possible that more than one antimicrobial substance is being produced by strain SCE2. PMID- 10389259 TI - Shiga toxin-producing Escherichia coli O157 in feedlot cattle and Norwegian rats from a large-scale farm. AB - A total of 365 faecal samples from different categories of cattle, 12 samples of untreated slurry, 50 samples of fresh droppings of feral domestic pigeons, 20 samples of fresh droppings of domestic sparrows and stool samples of 19 synanthropic rodents were examined for the presence of Escherichia coli by broth enrichment culture and a subsequent immunomagnetic separation. Escherichia coli O157 was found in 72 (20%) bovine samples, six (50%) samples of untreated slurry and four (40%) of 10 rats (Rattus norvegicus). Significant differences were found in the E. coli O157 shedding frequency between different age categories of bulls. Genes stx2 and eaeA were detected in all isolates, and the stx1 gene in all but 10 isolates. PMID- 10389260 TI - Characterization of Bacillus thuringiensis serovar bolivia (serotype H63), a novel serovar isolated from the Bolivian high valleys. AB - The type strain Bacillus thuringiensis var. bolivia (serotype H63), isolated from the Bolivian high valleys, has been characterized at different levels. Its parasporal crystal has an unusual shape and it is composed of a protein of 155 kDa which shows two bands of 75 and 80 kDa after activation. Analysis by PCR shows the presence of cry1 genes, and amplification with specific primers gave products for cry1 E, cry1 D, cry4 A and cry4 B with sizes different to those expected. Immunoblotting tests showed positive reaction for Cry1 E, Cry3 A, Cry4 A and Cry11 A crystal proteins. The plasmid pattern revealed two large and two small plasmids. Toxicity tests were performed against 14 insects and a slight toxicity was found against Plutella xylotella and Trichoplusia ni. PMID- 10389261 TI - An easy colorimetric assay for screening and qualitative assessment of deiodination and dehalogenation by bacterial cultures. AB - Halogenated organic substances are among the main environmental concerns. A number of micro-organisms are able to dehalogenate these compounds. However, the methods for the assessment of micro-organismal ability to dehalogenate are expensive and require complex instrumentation. Here, an easy colorimetric assay for the screening and assessment of the ability of bacterial cultures to deiodinate, and potentially dehalogenate, chemical substances is proposed. The method is based on the oxidation of iodide, released due to biotransformation, to iodine followed by a subsequent detection of iodine by a classical reaction with starch. PMID- 10389262 TI - Prevalence and antimicrobial susceptibility of staphylococci isolated from saliva of clinically normal cats. AB - Samples were collected from 150 adult cats, processed for isolation of Staphylococcus species and tested for susceptibility to penicillin G, gentamicin, oxacillin, enrofloxin and tetracycline. Methicillin resistance was also determined. One hundred and four isolates were obtained (69.3% of samples). Coagulase-negative species were most common, and the most frequently isolated (33 samples) species was Staph. felis. Other coagulase-negative species, such as Staph. haemolyticus, Staph. simulans, Staph. epidermidis and Staph. saprophyticus were also isolated. Coagulase-positive staphylococci were obtained from 30 cats, and the only species recovered in this group was Staph. intermedius. Resistance to antibiotics was frequently observed, with 68.2% of the isolates showing resistance to at least one drug. Resistance to Penicillin G was observed in 68 of the 104 isolates (65.4%), 23 samples were resistant to oxacillin (22.1%), 33 to tetracycline (31.7%) and 24 to enrofloxin (23.1%). Gentamicin was the most active antimicrobial agent. The role of these microorganisms in the saliva of cats is discussed. PMID- 10389263 TI - A novel double-membrane system for simultaneous nitrification and denitrification in a single tank. AB - A novel biological treatment system, which contains two types of membrane modules in a single tank, was developed for simultaneous nitrification and denitrification. Both of the modules were fed with the substrates on the tube side of the silicone tubes by diffusing them to the biofilms which form on the surface of the tubes. One module was fed with methanol for denitrification and the other one was fed with pure oxygen for nitrification. As a result, the interference of organic carbon on nitrification, and that of oxygen on denitrification, were both hindered by the diffusion barriers (biofilms), thereby allowing two different niches for nitrifiers and denitrifiers to coexist in a single tank. Besides saving space and the amount of alkalinity required for nitrification, this system also produced low residual chemical oxygen demand (COD) and high nitrogen removal rates (2.9-3.4 gN m-2 d-1 of surface area of membrane). PMID- 10389264 TI - Formation of a chiral acetoinic compound from diacetyl by Escherichia coli expressing meso-2,3-butanediol dehydrogenase. AB - L-Acetoin (L-AC) was produced from diacetyl (DA) by Escherichia coli JM109/pBUD119 containing the meso-2,3-butanediol dehydrogenase (D-AC forming) gene. However, when the strain was cultured in the presence of isopropyl-beta-D thiogalacto-pyranoside, the enzyme formed catalysed not only D-AC but also L-AC. L-AC was further reduced to L-2,3-butanediol (BD). The yield of L-AC or L-BD from DA (3 gl-1) was about 70% (w/w). PMID- 10389265 TI - Effect of higher pasteurization temperatures, and longer holding times at 72 degrees C, on the inactivation of Mycobacterium paratuberculosis in milk. AB - Raw cow's milk spiked with 10(6) cfu ml-1 of Mycobacterium paratuberculosis was subjected to heat treatments of 72, 75, 78, 80, 85 or 90 degrees C for 15 s, and 72 degrees C for 20 and 25 s, using laboratory pasteurizing units. Three bovine strains of Myco. paratuberculosis were studied (NCTC 8578, B2 and DVL 943). Each strain was subjected to all the heat treatments indicated on three separate occasions. Although each of the heat treatments achieved a substantial (5-6 log10) reduction in numbers of viable Myco. paratuberculosis, small numbers of the organism (4-16 cfu 10 ml-1) survived in a proportion of the milk samples at each of the higher temperatures investigated, right up to 90 degrees C for 15 s. A longer holding time of 25 s at 72 degrees C was found to be more effective at inactivating Myco. paratuberculosis. Only one of the three strains studied, B2, yielded small numbers of survivors after heating at 72 degrees C for 20 s, but it was completely inactivated by extending the holding time at 72 degrees C by a further 5 s to 25 s. It was concluded that a longer holding time is more likely to achieve the complete inactivation of Myco. paratuberculosis in milk than a higher pasteurization temperature. PMID- 10389266 TI - Prebiotic synthesis of vitamin B6-type compounds. AB - Heating a dilute solution of NH3 and glycoaldehyde gives a large family of pyridines substituted with the same functional groups as occur in the forms of vitamin B6. Thus, vitamin B6-like molecules could have been present on the early Earth and could have been available for catalysis of primitive transamination reactions. Ethanolamine and N-methylethanolamine are also formed as major products. These are choline-like molecules, the latter of which is apparently formed by a prebiotic methylation process. PMID- 10389268 TI - Highly oxygenated bisabolenes and an acetylene from Matricaria aurea. AB - Reinvestigation of the aerial parts of Matricaria aurea led to the isolation of three new bisabolenes and a new acetylene. The structures of the four compounds, namely (1R*,2R*,3R*,6R*,7R*)1,2,3,6,7- pentahydroxy-bisabol-10(11)-ene, (1R*,2R*,3R*,6R*,7R*)1,2,3,6,7-pentahydroxy-1-acetoxy-bisabol-10(1 1)-ene, (1R*,2R*,3R*,6R*,7R*)1,2,3,6,7-pentahydroxy-2-acetoxy-bisabol-10(1 1)-ene and (3S*,4S*,5R*)-(E)-3,4-dihydroxy-2-(hexa-2,4-diynyliden)-1,6- dioxaspiro (4,5)decane, were deduced from the high field NMR studies. PMID- 10389267 TI - Bioactive sesquiterpenes from Santolina rosmarinifolia subsp. Canescens. A conformational analysis of the germacrane ring. AB - The hexane extract of aerial parts of Santolina rosmarinifolia subsp. canescens afforded eight new sesquiterpenes in addition to known compounds. Their structures were determined by spectroscopic methods and chemical transformations. The conformational analysis of the germacrane constituents was carried out by spectroscopic methods, including NMR at varying temperature and by molecular mechanics calculations. The antifeedant, antibacterial and antitumoral activity of selected compounds has been tested. PMID- 10389269 TI - Steroidal saponins from the bulbs of Lilium candidum. AB - Five new spirostanol saponins and a new furostanol saponin were isolated from the fresh bulbs of Lilium candidum. Their structures were elucidated on the basis of spectroscopic analysis, including two-dimensional NMR spectroscopic techniques and the result of acid hydrolysis. The isolated saponins contained a branched triglycoside moiety assigned as O-alpha-L-rhamnopyranosyl-(1-->2)-O-[beta-D glucopyranosyl-(1-->6)]-beta - D-glucopyranose with the formation of an O glycosidic linkage to C-3 of the aglycone as the common structural feature. The inhibitory activity of the saponins on Na+/K+ ATPase was evaluated. PMID- 10389270 TI - Flavanone glycosides from Alhagi pseudalhagi. AB - Two new flavanone glycosides, alhagitin and alhagidin, have been isolated from the whole plant of Alhagi pseudalhagi and their structures established respectively as naringenin 5-methyl ether 4'-glucoside and hesperitin 7 galactosyl(1-->2)[rhamnosyl(1-->6)]glucoside by chemical and spectroscopic methods. PMID- 10389271 TI - Isolation and partial characterisation of galactose-specific lectins from African yam beans, Sphenostyles stenocarpa Harms. AB - A new galactose-specific lectin was isolated from African yam bean (Sphenostyles stenocarpa Harms) by affinity chromatography on galactose-Sepharose 4B. SDS-PAGE analysis resulted in four polypeptide bands of approximately 27, 29, 32 and 34 kDa, respectively. Based on the analysis of carbohydrate content and native PAGE, it is likely that the Sphenostyles lectin is a tetrameric glycoprotein with M(r) of approximately 122 kDa. N-terminal protein sequencing of purified lectins from four different Sphenostyles accessions shows that the four polypeptides have largely identical amino acid sequences. The sequences contain the conserved consensus sequence F-F-LILG characteristic of legume lectins, as well as Phaseolus vulgaris proteins in the arcelin-alpha-amylase inhibitor gene family. The lectin agglutinates both rabbit and human erythrocytes, but with a preference for blood types A and O. Using Western blotting, the lectin was shown to accumulate rapidly during seed development, but levels dropped slightly as seeds attained maturity. This is the first time a lectin has been purified from the genus Sphenostyles. The new lectin was assigned the abbreviation LECp.SphSte.se.Hga1. PMID- 10389273 TI - Biotransformation of the diperpenoid, isosteviol, by Aspergillus niger, Penicillium chrysogenum and Rhizopus arrhizus. AB - The biotransformation of isosteviol (ent-16-ketobeyeran-19-oic acid) by three fungi is described. Aspergillus niger produced the 7 beta-OH derivative, ent-7 alpha-hydroxy-16-ketobeyeran-19-oic, and the 1 alpha, 7 beta-diOH derivative, ent 1 beta, 7 alpha-dihydroxy-16-ketobeyeran-19-oic acid. The 17-OH compound, ent-17 hydroxy-16-ketobeyeran-19-oic acid, was obtained with Penicillium chrysogenum. Rhizopus arrhizus produced the 7 beta-OH derivative, ent-7 alpha-hydroxy-16 ketobeyeran-19-oic acid. The isolated metabolites were characterised by IR, NMR and MS. PMID- 10389272 TI - Bioactive N-isobutylamides from the flower buds of Spilanthes acmella. AB - The hexane extract of dried flower buds of Spilanthes acmella afforded three N isobutyl amides: spilanthol, undeca-2E,7Z,9E-trienoic acid isobutylamide and undeca-2E-en-8,10-diynoic acid isobutylamide. Their structures were determined by 1H and 13C NMR, MS and GC-MS spectroscopic methods. All were active against Aedes aegyptii larvae and Helicoverpa zea neonates at 12.5 and 250 micrograms/mL concentrations, respectively. PMID- 10389274 TI - Iridoid and seco-iridoid glucosides from Chioccoca alba (Rubiaceae). AB - Phytochemical investigation of Chioccoca alba afforded three new iridoids, alboside I, alboside II and alboside III, and a new seco-iridoid alboside V. Alboside IV showed moderate activity towards the DNA repair-deficient mutant RS321 of Saccharomyces cerevisiae. The structural elucidation of the new compounds was performed by ES-MS and by 1D and 2D NMR spectroscopic methods. PMID- 10389275 TI - Limonoids from fruit of Melia toosendan and their cytotoxic activity. AB - Two new limonoids, toosendanal and 12-O-methylvolkensin, were isolated from the fruits of Melia toosendan Sieb. et Zucc. along with three known limonoids, meliatoxin B1, trichillin H, and toosendanin. The structures of the new limonoids were established by spectroscopic methods, with toosendanal having C-1/C-29 and C 19/C-29 acetal bridges. Both meliatoxin B1 and toosendanin exhibit cytotoxic activity against KB cells. PMID- 10389276 TI - Phenethyl alcohol glycosides and isopentenol glycoside from fruit of Bupleurum falcatum. AB - Investigation on the constituents of the fruit of Bupleurum falcatum L. resulted in the isolation of the three new glycosides, phenethyl alcohol 8-O-beta-D glucopyranosyl-(1-->2)-O-beta-D-apiofuranosyl-(1-->6)-b eta-D- glucopyranoside, phenethyl alcohol 8-O-beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranoside and isopentenol 1-O-beta-D-apiofuranosyl-(1-->6)-beta-D-glucopyranoside along with five known glycosides, icariside D1, icariside F2, saikosaponin a, saikosaponin c and saikosaponin d. The structures of these compounds were elucidated on the basis of interpretation of chemical and spectral data. PMID- 10389278 TI - Towards single atom analysis of biological structures. AB - Mapping single atoms in biological structures is now becoming within the reach of analytical electron microscopy. Electron energy-loss spectroscopy (EELS) in the field-emission scanning transmission electron microscope (STEM) provides a particularly high sensitivity for detecting the biologically important element, phosphorus. Imaging can be performed at low dose with dark-field STEM prior to analysis at high dose, so that structures of macromolecular assemblies can be correlated with the numbers of specific atoms that they contain. Measurements confirm theoretical predictions that single atom detection requires a nanometer sized probe. Although phosphorus atoms may have moved several nanometers from their original positions by beam-induced structural degradation at the high required dose of approximately 10(9) e/nm2, damaged molecules are nevertheless stable enough to be analyzed at 1 or 2 nm resolution. Such analyses can only be achieved by means of spectrum-imaging with correction for specimen drift. Optimal strategies for mapping small numbers of phosphorus atoms have been investigated using well-characterized specimens of DNA plasmids and tobacco mosaic virus. PMID- 10389279 TI - Computerized tomographic angiography in the evaluation of cerebral infarction. AB - BACKGROUND: To evaluate the feasibility of computerized tomographic angiography (CTA) for the diagnosis of cerebral infarction. METHODS: Fourteen patients (nine men, five women; average age, 60 years) who presented with symptoms of acute cerebral infarction underwent computerized tomography (CT) and CTA, using a Picker PQ 2000 Spiral CT. The infarcted area was first evaluated by CT and then occlusive sites of arteries were evaluated by CTA. RESULTS: In seven patients in the very early stage of acute cerebral infarction involving the middle cerebral artery (MCA) territory, the initial CT did not clearly demonstrate the infarcted area. CTA detected the occlusive site of the MCA and helped anticipate the area of infarction. Follow-up CT confirmed the infarction clearly. CTA was not useful for small infarctions in the basal ganglia, thalamus or brainstem. CONCLUSIONS: CTA in association with CT is a feasible and good technique for the diagnosis of early-stage acute cerebral infarction. It can detect the occlusive sites in the main arteries, especially the middle cerebral artery, and as of benefit for anticipation of the ensuing edematous area. However, for small infarcts in the basal ganglia, CTA does not provide good diagnostic images. PMID- 10389280 TI - Estimation of costs due to hospitalization for first-ever stroke patients in northern Taiwan. AB - BACKGROUND: The need for healthcare services and the related costs for stroke patients may rise steadily in the future. Even with the predictable and substantial burden of stroke, little effort has been devoted to measuring the population-based direct medical and nonmedical costs in Taiwan. METHODS: Data from the study "Epidemiological Study of Stroke, Diabetes, and Cardiovascular Disease," which included 8,705 people older than 35 years of age, and the study "Costs of Stroke," which included 660 first-ever stroke patients, were used for the cost calculations. The cost of hospital care for stroke patients was obtained in two steps. First, the incidence of stroke and readmissions within one year were tallied; the sum was then multiplied by the average length of stay. Second, the total medical and nonmedical costs were divided by the sum obtained from step 1. The resulting quotient obtained was the cost of hospital care for stroke patients per day. RESULTS: There were 6,691 incidents of stroke and stroke related readmissions in 1995 (4,041 men and 2,650 women). The total person-days of hospital stay were 233,569 days (144,264 for men and 89,305 days for women). The average medical and nonmedical costs of hospital care per person-day was US $251.4 (NT $6,788 at an exchange rate of US $1 = NT $27). Cost for men (US $287, NT $7,749) was more than for women (US $208, NT $5,616). The total direct costs of hospital care were US $58,710,000 (NT $1,585,000,000) in 1995. CONCLUSIONS: An average of US $1,682,000 (NT $45,410,000) in hospital care costs for stroke could have been saved in 1995 if the person-day stay had been decreased by only one day. PMID- 10389281 TI - Natural course of submacular hemorrhage. AB - BACKGROUND: Age-related macular degeneration (ARMD) is the most prevalent cause of blindness in the elderly population. This study retrospectively evaluated the natural course of submacular hemorrhage related to ARMD. METHODS: We reviewed the records of patients with submacular hemorrhage and foveal avascular zone involvement at the Veterans General Hospital-Taipei from 1981 to 1996. Data were collected from color fundus pictures, fluorescence angiographic films and visual acuity changes. RESULTS: Data from a total of 86 eyes with age-related macular degeneration (ARMD) and 37 non-ARMD eyes were collected. Analysis of data from the ARMD group revealed 10.5% visual acuity improvement six months after presentation and a final mean acuity of 0.069. Analysis of data from the non-ARMD group revealed 29.7% visual acuity improvement and a final mean acuity of 0.388. CONCLUSIONS: In this study, patients with submacular hemorrhage had spontaneous visual improvement even without surgery, especially those without subretinal neovascularization. PMID- 10389282 TI - Pilocytic astrocytoma of the posterior fossa: a follow-up study in 15 patients. AB - BACKGROUND: The extent of resection in pilocytic astrocytoma of the posterior fossa remains undefined, as the problem of hydrocephalus has not yet been solved. We retrospectively reviewed the data from 15 patients with a pilocytic astrocytoma of the posterior fossa to evaluate the impact of surgical technique, in terms of resection extent, by serial magnetic resonance imaging (MRI) examinations. In addition, the issue of hydrocephalus was considered and related to the different treatment modalities. METHODS: Macroscopic, gross, total resection of the tumor was performed in all 15 patients. Follow-up was obtained in 14 patients for a period ranging between 11 and 119 months (median, 41.5 months). The ages of patients ranged from two to 13 years (mean, 7 +/- 3 years). All patients underwent serial MRI examinations in the first month, every six months for the first two years and then yearly. RESULTS: Outcome was good in 12 patients who had no neurologic deficit and fair in two patients who were slightly handicapped but had an independent life. There were four patients with an abnormally persistent enhancement on MRI, with a median follow-up of 30 months. One of these patients had progressively increasing size of the enhancement. Three of them had the same size of enhancement during the follow-up period. The MRI findings showed residual tumors in four patients. One of them had tumor regrowth one year after surgery. There were 11 cases with pilocytic astrocytoma and hydrocephalus. Five patients were treated with tumor removal and external CSF drainage. Six patients underwent tumor removal only, without perioperative cerebrospinal fluid (CSF) drainage. Only one patient had a permanent ventriculoperitoneal shunt. CONCLUSIONS: Our study illustrated that the extent of tumor resection of pilocytic astrocytoma can be defined by postoperative serial MRI examinations. Long-term follow-up with MRI seems mandatory in cases with abnormal enhancement. Hydrocephalus is a common finding in patients with a pilocytic astrocytoma. A permanent ventriculoperitoneal shunt is required only in patients with postoperative hydrocephalus. PMID- 10389283 TI - Peri-anhepatic phase oxygen kinetics in porcine liver transplantation. AB - BACKGROUND: We applied a liver transplantation animal model to examine the relationship between oxygen delivery and consumption. The presence of pathologic flow-dependent oxygen consumption was investigated during and after the anhepatic phase. The effect of venous-to-venous bypass on oxygen kinetics was evaluated. METHODS: Twelve pigs were randomly divided into two groups. The non-bypass group consisted of six pigs that were subjected to clamping of the hepatic artery, portal vein, and the superior and inferior vena cava to produce an anhepatic phase. The bypass group consisted of six pigs that underwent vascular clamping and liver transplantation with venous bypass. Hemodynamics, oxygen delivery index (DO2) and oxygen consumption index (VO2) were recorded during the peri-anhepatic phase. Best-fit regression lines were calculated for DO2 vs VO2. RESULTS: In the pigs without venous bypass, the blood pressure, cardiac index and VO2 dropped significantly after vascular clamping and lactic acidosis developed. In pigs with venous bypass, vascular clamping induced a significant decline of cardiac output and DO2 but VO2 was maintained by a compensatory increase in oxygen extraction ratio. DO2 and VO2 after the release of vascular clamping increased significantly higher than that before vascular clamping. The O2 supply-dependent regression line was drawn from the points below critical oxygen delivery with a slope of 0.232 (95% CI = 0.110-0.354, r2 = 0.50, p = 0.010). The pathologic supply dependent line was drawn from the points with supranormal DO2 and VO2 with a slope of 0.185 (95% CI = 0.050-0.333, r2 = 0.510, p = 0.029). The slope of the supply-independent line was 0.0089 (95% CI = -0.030-0.050, r2 < 0.009, p = 0.12). CONCLUSIONS: Oxygen delivery dropped below the critical level and flow-dependent oxygen consumption developed during the anhepatic phase without venous bypass. Venous-to-venous bypass is necessary to maintain a critical DO2 and stable hemodynamics during porcine liver transplantation. Pathologic flow-dependent oxygen consumption developed after the anhepatic phase. PMID- 10389284 TI - Relationship between hormone receptor concentration and tumor shrinkage in uterine myoma after treatment with a GnRHa. AB - BACKGROUND: Uterine myomas are benign tumors of the uterus, occurring in up to 25% of women of reproductive age. We examined the possible causes for different degrees of volume reduction in patients with uterine myomas who received gonadotropin-releasing hormone agonist (GnRHa) treatment by investigating the hormone receptors at the end of GnRHa treatment. METHODS: This trial was designed as a prospective study of five premenopausal women presenting with symptomatic uterine myoma. All patients were treated with a subcutaneous injection of goserelin depot 3.6 mg every four weeks for 16 weeks. Clinical examinations, hormonal evaluation and ultrasound determinations were performed before, during and after treatment. At the end of the treatment period, all patients underwent myomectomy. The concentrations of the unbound progesterone receptors and estrogen receptors were evaluated. RESULTS: The volume of the uterine myoma decreased by 21% to 65%. The percentage of decrease in volume of treated uterine myomas was found to negatively correlate with the concentration of unbound progesterone receptors (r2 = 0.92, p = 0.008). This percentage was not significantly correlated with the concentration of unbound estrogen receptors (r2 = 0.02, p = 0.84). CONCLUSIONS: The limited data available suggested that the volume decrease of uterine myomas in GnRHa-treated patients is partly dependent on the concentrations of unbound sex-hormone receptors in the uterine myomas. PMID- 10389285 TI - Pseudomonas aeruginosa central nervous system infections: analysis of clinical features of 16 adult patients. AB - BACKGROUND: The purpose of this study was to analyze the clinical features and therapeutic outcome of 16 adult patients with Pseudomonas aeruginosa central nervous system (CNS) infection. We also attempted to identify the factors that significantly influence the prognosis of this potentially fatal CNS infection. METHODS: Sixteen adult patients with P aeruginosa CNS infection, nine men and seven women, aged from 18 to 86 years, were included in this retrospective study. The clinical features and the laboratory data of these patients were analyzed. Potential prognostic factors were compared by means of Fisher's exact test and the relative risks were estimated by odds ratio. RESULTS: Of the 16 patients, 13 had meningitis and three had focal suppuration (two with brain abscess and one with spinal epidural abscess). The 13 meningitis patients with nosocomial or community-acquired infections were classified into two forms: the spontaneous form and the neurosurgical form. The overall mortality rate was 37.5% (6/16). In the meningitis group, the patients with the neurosurgical form had a lower mortality rate (11.1%; 1/9) than the patients with the spontaneous form (100%; 4/4), and those with community-acquired meningitis had a higher mortality rate (80%; 4/5) than those with nosocomial infections (12.5%; 1/8). All the meningitis patients who did not receive appropriate antibiotic treatment expired. The statistically significant prognostic factors included the acquisition of infection, form of infection, bacteremia, initial level of consciousness and the use of appropriate antibiotics. CONCLUSIONS: Vigilance for P aeruginosa is particularly important in patients with predisposing factors such as head injury, neurosurgical procedures and long-term debilitating diseases. Early appropriate antibiotic therapy and neurosurgical intervention for patients with suppurative infections can bring about improved therapeutic results. PMID- 10389286 TI - Evaluation of conjugated estrogen plus medroxyprogesterone acetate versus tibolone in early postmenopausal Chinese women. AB - BACKGROUND: The safety and efficacy of tibolone (Livial) were compared with the traditional cyclic, sequential conjugated estrogens/medroxyprogesterone acetate (Premarin/Provera; PP) regimen for the treatment of climacteric symptoms, prevention of postmenopausal bone loss, endometrial stimulation and influence on lipid profile. METHODS: Forty women, one to three years postmenopause, were randomly enrolled in one of two treatment groups, receiving either tibolone (2.5 mg) every day for six months or Premarin (0.625 mg) every day plus Provera (5 mg) from day 1 to day 12 every month for six months. The scores of climacteric complaints, using the Greene Climacteric Scales, and bleeding pattern were recorded at baseline and follow-up visits at months 1, 3 and 6. Bone resorption (deoxypyridinium) and formation (osteocalcin) markers were measured at baseline, three and six months. Lipid profiles, bone density of the lumbar spine and neck of the femur measured by dual energy X-ray absorptiometry were checked at baseline and six months. RESULTS: Tibolone was as effective as PP in alleviating climacteric complaints. Both regimens were effective in slowing bone metabolism and preventing bone loss. After six months of treatment, bone density of the lumbar spine increased 2.174% in the tibolone group. The endometrium of patients remained atrophic (< 4 mm); only one woman reported vaginal spotting after three months of tibolone therapy. Significant decreases in triglyceride (31.48%) and high-density lipoprotein (29.25%) were also observed. In the PP group, bone density of the lumbar spine increased 1.405%; cyclic withdrawal bleeding occurred in every patient. A significant increase in triglyceride (38.76%) and a significant decrease in low-density lipoprotein (15.10%) were observed. CONCLUSIONS: Tibolone proved to be effective and safe in the treatment of women with climacteric symptoms and postmenopausal bone loss. As a form of hormone replacement therapy without the need for withdrawal bleeding, tibolone has great appeal to postmenopausal women, and compliance is higher than reported with other forms of hormone replacement therapy. PMID- 10389287 TI - Progression of cytomegalovirus retinitis in acquired immunodeficiency syndrome: a case report. AB - We report an AIDS patient with cytomegalovirus (CMV) retinitis that developed from an early minor lesion and progressed to extended involvement of the retina and severe deterioration of vision due to poor compliance with ganciclovir treatment. A 33-year-old man was known to have acquired immunodeficiency syndrome (AIDS) for eight months. The patient had no complaint of visual symptoms. A routine eye examination revealed his visual acuity to be 6/6 in both eyes. The dilated eye fundus examination using indirect ophthalmoscopy disclosed a localized white yellowish granular lesion in the peripheral retina of the right eye and a completely normal left eye. CMV retinitis with initial manifestation in the right eye was diagnosed. Due to incomplete treatment with ganciclovir, the retinal lesion rapidly enlarged and extended to the posterior pole, with eventual destruction of the nerve fiber layer and optic disc. The visual acuity of right eye dropped from 6/6 to 1/60 within six months. This case report indicates the importance of early, dilated eye fundus examination and recognition of early CMV retinitis in order to salvage visual function in AIDS patients. Completion of the anti-CMV treatment course in halting the progression of CMV retinitis is also emphasized. PMID- 10389288 TI - Diagnosis and treatment of cystic lymphangioma of the ascending colon by laparoscopic-assisted surgery: a case report. AB - Lymphangioma of the colon is a rare disease. Its clinical silence and absence of specific symptoms and signs make it difficult to diagnose preoperatively. We present a case of cystic lymphangioma of the ascending colon associated with constipation in a 72-year-old man and review the pertinent literature. The patient underwent laparoscopic-assisted segmental resection of the colon. The characteristic histologic appearance of cystic lymphangioma provided the definitive diagnosis. The recovery course was uneventful. Two years postoperatively, the patient was symptom-free and without evidence of tumor recurrence. PMID- 10389289 TI - Serum levels of cytokines in hepatitis C-related liver disease: a longitudinal study. AB - BACKGROUND: Elevated serum cytokine levels are found in patients with acute and chronic hepatitis B. However, little is known about the development and progression of cytokines in hepatitis C infection. We conducted this study to evaluate the change and clinical significance of cytokines in the different stages of hepatitis C infection. METHODS: Circulating interleukin-1 beta (IL-1 beta), interleukin-2 receptor (IL-2r), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) were measured by enzyme-linked immunosorbent assay in 29 patients with acute hepatitis C (AHC), 43 patients with chronic hepatitis C (CHC), 40 patients with liver cirrhosis (LC) and positive serum anti-hepatitis C antibody (anti-HCV), 36 patients with hepatocellular carcinoma (HCC) and positive anti-HCV and 30 normal controls. A cohort of patients with chronic hepatitis C was monitored for a median of seven years. RESULTS: Serum IL-1 beta, IL-2r, IL-6 and TNF-alpha levels were significantly elevated in all patient groups compared with controls (p < 0.05). The serum IL-1 beta, IL-2r and TNF-alpha levels in patients with LC or HCC were higher than that in patients with AHC or CHC (p < 0.05). In the longitudinal follow-up, 12 patients with chronic hepatitis at enrollment in the study developed liver cirrhosis. For these patients, serum levels of IL-1 alpha, IL-2r and TNF-alpha were higher in liver cirrhosis than in chronic hepatitis (p < 0.05). In addition, the serum concentrations of these cytokines correlated better with indices of hepatic dysfunction (prothrombin time and indocyanine green retention ratio) than with parameters of hepatic inflammation (alanine aminotransferase and aspartate aminotransferase). CONCLUSIONS: Serum IL-1 beta, IL-2r, IL-6 and TNF-alpha levels are elevated in patients with hepatitis C-related liver diseases, especially in LC and HCC patients. These levels reflect hepatic dysfunction better than liver inflammation parameters, which might explain the higher serum concentrations of cytokines in LC patients. PMID- 10389290 TI - Circadian and weekly variations in pain onset of acute myocardial infarction. AB - BACKGROUND: A seasonal variation with more myocardial infarctions in the winter months due to cold weather has been reported. Other reports have described excess numbers of acute myocardial infarction (AMI) in the summer in Southern USA due to hot temperatures. To determine whether circadian and seasonal variations affect the incidence of AMI in the subtropical area of Taiwan, 480 consecutive patients with AMI admitted to our coronary care units were analyzed. METHODS: Six-hourly intervals over 24 hours (4 periods), daily intervals in a week (7 days) and monthly intervals in a year (12 months) were respectively studied. The distributions of the numbers of AMI occurring in the six-hour intervals were tested for differences among the four periods using the chi-squared test for goodness of fit. RESULTS: We found that there was a circadian variation in the onset of AMI with a morning peak (6 am to noon) (35%, chi 2 = 28.52, df = 3, p < 0.01) but no secondary late evening peak. The incidence of AMI was significantly lower on Sundays (9%) than on the other weekdays (chi 2 = 16.37, df = 6, p = 0.012). However, no seasonal variation (no winter or summer peaks) occurred in the incidence of AMI in this study (chi 2 = 0.77, df = 3, p = 0.99). CONCLUSION: Our results showed that there was a predominant morning peak in the onset of AMI. The low incidence of AMI cases on Sunday compared with other weekdays suggested that relief from tension or workload on Sundays might have an important role in this low percentage of AMI. Differing from other reports, there was no seasonal variation in the occurrence of AMI in our study, suggesting that the warm climate of a subtropical region does not provide an environment that is likely to increase the frequency of AMI. PMID- 10389291 TI - Computerized tomography-guided stereotactic aspiration of brain abscesses: experience with 28 cases. AB - BACKGROUND: Computerized tomography (CT)-guided stereotactic techniques allow accurate identification of brain abscesses and provide promising results for the management of brain abscesses. METHODS: We reviewed the results of stereotactic aspiration of brain abscesses in 28 consecutive patients from 1984 to 1995. In all patients, the diagnosis of brain abscess was made by computerized tomography (CT). All patients underwent stereotactic aspiration of abscesses as the primary surgical therapy. Intravenous antibiotics were administered preoperatively and adjusted according to organism type and sensitivity to antibiotics. In patients with multiple lesions, aspirations were performed on abscesses larger than 2 cm in diameter or on those causing significant mass effects. CT was performed weekly to monitor abscess growth or failure to resolve. Patients were followed on an outpatient basis. This report is a retrospective review of clinical features, diagnostic methods, treatment and postoperative results. RESULTS: A total of 19 patients had good recoveries and six patients had mild neurologic sequelae. One patient had persistent conscious impairment. Intracranial hemorrhage occurred in one patient. Two deaths occurred during hospitalization. One patient with a fungal infection underwent additional surgical excision of the abscess. Most patients had resolution of abscesses after stereotactic treatment within two months. The cure rate was 92% in patients with bacterial brain abscesses treated with stereotactic aspiration and intravenous antibiotics for six weeks. CONCLUSIONS: Stereotactic surgery is a procedure with minimal morbidity and mortality, and can be the treatment of choice for brain abscesses when combined with appropriate antibiotic therapy. PMID- 10389292 TI - Detection of IgA against Epstein-Barr virus BZLF-1 replication activator (ZEBRA) in sera of nasopharyngeal carcinoma patients with a recombinant ZEBRA protein. AB - BACKGROUND: Epstein-Barr virus (EBV) is closely associated with nasopharyngeal carcinoma (NPC). An EBV-encoded immediate-early antigen, BZLF-1 replication activator (ZEBRA) initiates EBV replication and expression in all NPC tumors. In this study, we investigated whether immunoglobulin A (IgA) against ZEBRA is present in the sera of patients with NPC, and whether it was able to be determined by enzyme-linked immunosorbent assay (ELISA) using a recombinant ZEBRA prepared from Escherichia coli. METHODS: A polymerase chain reaction-amplified cDNA fragment of the ZEBRA gene was inserted into the expression vector of E coli under the control of an IpL promoter. E coli bacteria containing the CI857 gene served as host to overexpress the ZEBRA protein by heat induction. Recombinant ZEBRA was collected by mechanical disruption of the bacteria, purified by column chromatography, and analyzed by SDS-PAGE and Western blot assay using sera from NPC patients. The recombinant ZEBRA was used to develop the ELISA to detect IgA against ZEBRA. RESULTS: The amount of ZEBRA produced comprised 30% of total E coli protein. Western blot assay confirmed that affinity of the recombinant ZEBRA to IgA antibody was preserved. IgA against ZEBRA was shown to be positive by ELISA in 36 of 40 NPC sera, but in only nine of 55 patients with other head and neck malignancies, and two of 35 normal individuals. For serologic diagnosis of NPC, the sensitivity of IgA/ZEBRA detected by ELISA was 90% and the specificity was 87.4%. CONCLUSIONS: A recombinant ZEBRA was produced at high levels in E coli and retained affinity to IgA against ZEBRA. The recombinant ZEBRA was successfully used to develop an ELISA for the detection of IgA against ZEBRA. The high sensitivity and specificity of IgA against ZEBRA show that the ELISA is feasible for serologic diagnosis of NPC. PMID- 10389293 TI - Observations after orchiectomy in clinical stage I nonseminomatous germ cell tumors of the testis. AB - BACKGROUND: The optimal management of clinical stage I nonseminomatous germ cell tumor (NSGCT) of the testis remains controversial. For years, retroperitoneal lymph node dissection in combination with orchiectomy, has been the standard treatment in patients with clinical stage I NSGCT. Recently, with advancement of effective cisplatin-based chemotherapy and clinical staging procedures, a new approach of observation after orchiectomy is being evaluated. We reviewed cases of orchiectomy and observation for clinical stage I NSGCT of the testis in order to evaluate the treatment outcome. METHODS: We retrospectively reviewed the records of 13 patients with clinical stage I NSGCT of the testis treated at our hospital from February, 1981 to August, 1996. The patient age at diagnosis ranged from 0.6 to 44 years. Nine patients had yolk sac tumors, and four had mixed germ cell tumors. Median follow-up was 42 months (range, 20-132 months). RESULTS: Prior to orchiectomy, serum beta-human chorionic gonadotropin and alpha fetoprotein (AFP) were raised to abnormal concentrations in four and in 13 patients, respectively. With a median follow-up of 42 months, three of 13 patients relapsed at a median of three months after orchiectomy. Two patients showed elevated AFP and radiographically identifiable tumors simultaneously, and one patient showed elevated AFP as the only evidence of relapse. Following treatment with cisplatin-based chemotherapy, the three patients who relapsed responded successfully and the elevated AFP returned to normal. The patients are currently alive and disease free. CONCLUSIONS: Observation after orchiectomy is a reasonable approach for patients with clinical stage I NSGCT of the testis. PMID- 10389294 TI - Malignant otitis externa. AB - BACKGROUND: Malignant otitis externa is an infrequent but severe infection of the external auditory canal, most often affecting elderly diabetic patients. Early diagnosis is necessary due to its high morbidity and mortality. METHODS: From 1990 to 1997, all patients with malignant otitis externa at the Veterans General Hospital-Taipei were reviewed retrospectively. The clinical features and the strategy of diagnosis and treatment are discussed. RESULTS: Twelve patients with an average age of 65.3 years were included. Eleven of these patients were diabetic. All had the presenting symptoms of otalgia and otorrhea at diagnosis. Bacterial cultures grew Pseudomonas aeruginosa in eight patients and methicillin resistant Staphylococcus aureus in four patients. The mean duration of admission was 82 days. Appropriate antibiotics were given according to the results of bacterial culture and sensitivity test. 99Technetium scans and 67gallium scans were performed to evaluate the extent of involvement and monitor the effects of treatment. Eventually, four patients died due to renal failure, meningitis, pneumonia and upper gastrointestinal bleeding, respectively. CONCLUSIONS: Malignant otitis externa is a life-threatening infection arising from the external auditory canal. A high degree of suspicion for malignant otitis externa is mandatory. Vigorous local and systemic antimicrobial treatment should be initiated early in the course of the disease to achieve a satisfactory outcome. 99Technetium and 67gallium scans are important for the diagnosis and evaluation of the treatment results. PMID- 10389295 TI - Characteristics and risk factors of acetaminophen-induced hepatitis in Taiwan. AB - BACKGROUND: Overdose of acetaminophen may cause hepatic injury and fatal fulminant hepatic failure. Acetaminophen is the most common form of drug-induced hepatic injury in Western countries. However, there is no formal report of this important issue in Taiwan. To assess the clinical characteristics and risk factors of acetaminophen-induced hepatitis in Taiwan, we conducted this study. METHODS: A total of 71 patients who were intoxicated or overdosed with acetaminophen at Veterans General Hospital-Taipei between February, 1991 and June, 1997 were enrolled in this study. Acetaminophen-induced hepatitis was defined according to the Paris international consensus criteria. RESULTS: Suicide attempt was the major cause (67/71) of acetaminophen overdose. Sixty-two of the patients were female. The mean patient age was 24.5 +/- 9.7 years (+/- standard deviation, SD). Nineteen of 71 patients had acetaminophen-induced hepatitis. The peak serum alanine aminotransferase, aspartate aminotransferase and total bilirubin concentrations were 4,181.0 +/- 931.4 IU/l (mean +/- SD), 4,148.0 +/- 1,147.5 IU/l and 2.6 +/- 0.4 mg/dl, respectively. The hepatitis group had higher ingested acetaminophen doses and serum peak acetaminophen levels than did those of the nonhepatitis group (24.9 g vs 12.9 g, p = 0.004; 132.0 mg/l vs 61.7 mg/l, p = 0.013). A higher percentage of alcohol consumption was also noted in the hepatitis group than in the nonhepatitis group (32% vs 12%, p = 0.05). After logistic regression for multivariate analysis, alcohol consumption was the most important risk factor for acetaminophen-induced hepatitis (odds ratio = 8.14, p = 0.018), followed by ingested acetaminophen dose (odds ratio = 1.21, p = 0.001). Most patients received acetylcysteine treatment in time. Two of the 19 patients with hepatitis died. CONCLUSIONS: The majority of acetaminophen-induced hepatitis in Taiwan occurs in young females who attempt suicide. Alcohol consumption and the dose of acetaminophen ingested were the significant risk factors for acetaminophen-induced hepatitis. Alcohol should not be concomitantly used with acetaminophen. Early diagnosis and administration of the antidote are crucial to decrease mortality. PMID- 10389296 TI - Hemostatic changes in patients with liver cirrhosis. AB - BACKGROUND: Hemostatic changes in liver disease are complicated. An overall evaluation of the main hemostatic parameters in patients with different degrees of cirrhosis of the liver has not been reported in Taiwan. METHODS: A series of hemostatic tests and parameters including activated partial thromboplastin time, prothrombin time, thrombin time, bleeding time, factor VIII assay, antithrombin activity, fibrinogen, plasminogen, protamine sulfate test, fibrin(ogen) degradation products, D-dimer, thrombin-antithrombin complex (measured by modified antithrombin), tissue plasminogen activator (tPA), plasminogen activator inhibitor-1, euglobulin lysis test and venous occlusion test were performed in 51 patients with cirrhosis of the liver and 33 healthy controls. Among the cirrhotics, 18 were classified as Child-Pugh group A, 16 were B and 17 were C. RESULTS: Plasminogen, antithrombin and platelet count decreased progressively, starting with group A, then B and then C, relative to the controls. Factor VIII, activated partial thromboplastin time, prothrombin time, bleeding time, D-dimer and fibrin(ogen) degradation products increased progressively starting with group A, to B and then C, relative to controls. Severity of cirrhosis correlated with hemostatic changes. No significant change in the fibrinolytic response after challenge with the venous occlusion test was found in either Child-Pugh groups A, B, C or the controls, though progressive increases in tPA were found starting with group A, to B and then C, relative to controls. CONCLUSIONS: Our study proved a close relationship between the severity of cirrhosis and hemostatic changes. Activated partial thromboplastin time was better than bleeding time or thrombin time to demonstrate the severity of liver damage and hemostatic change in cirrhosis. Because the deterioration of coagulation function and increased fibrinolytic activity paralleled the severity of liver cirrhosis, adequate treatment for cirrhotic bleeding should not only correct the coagulation defects, but should also lower the increasing fibrinolytic activity. PMID- 10389297 TI - Term pregnancy in a noncommunicating rudimentary horn of an unicornuate uterus: a case report. AB - A case of full-term pregnancy in the noncommunicating rudimentary horn of unicornuate uterus is presented. A healthy female infant weighing 2,985 g was delivered by cesarean section at 37 weeks' gestation. The diagnosis was missed by prenatal ultrasonography and was made only at laparotomy. A retrospective analysis of our ultrasound studies revealed that the echo pattern of the nongravid uterus resembled a soft pelvic mass. Color Doppler imaging revealed prominent low-impedance uterine arcuate-radial arterial blood flow surrounding the periphery of the pelvic mass, as well as multiple areas of placental implantation with a pulsatile lacunar flow pattern. A high index of suspicion of rudimentary horn pregnancy should be borne in mind whenever late-pregnancy ultrasonography shows the above-mentioned characteristic ultrasonic findings. PMID- 10389298 TI - Coronal approach for the replacement of the condylar head in bilateral temporomandibular joint ankylosis: report of three cases. AB - Limitation of mouth opening has long been the chief complaint for patients who suffer from temporomandibular joint (TMJ) ankylosis. For surgical treatment of this disease, several ways have been employed for access to the condylar fossa, including the preauricular, postauricular, perimeatal, endaural and Risdon approaches. In this article, we report three patients with bilateral TMJ ankylosis who underwent replacement of both condylar heads using the bicoronal approach. The advantages demonstrated with this surgical technique are ease of access to the condylar head, ideal surgical exposure field, utility of temporalis muscle and fascia and minimal risk of facial paralysis. PMID- 10389299 TI - [Are ultraviolet and visible spectroscopy and spectrophotometry obsolete methods in pharmaceutical analysis?]. AB - It has been investigated if UV-VIS spectroscopy and spectrophotometry can be regarded to be obsolete methods in pharmaceutical analysis. The conclusions are as follows. As a consequence of the introduction and spreading of highly efficient spectroscopic methods in the structural analysis of organic compounds the importance of UV-VIS spectroscopy as a structure elucidation tool has greatly decreased. At the same time, however, diode-array UV spectrophotometers used as HPLC detectors have created very convenient possibilities for the identification of minor components (impurities, degradation products, etc.) in drugs. This statement is illustrated by several practical examples. On the basis of some data taken from the British Pharmacopoeia 1998 it is stated that UV spectrophotometry as a quantitative analytical method still belongs to the most frequently used analytical techniques in pharmaceutical analysis. At the same time, however, the authors are of negative opinion about the up-to-dateness and usefulness of colorimetric methods still very often published for the determination of drug substances. PMID- 10389300 TI - [Effect of immunosuppressive agents and antilymphocyte serum on bacterial translocation in mice]. AB - Following intraperitoneally applied treatment with 0.5 ml of ana partes diluted antilymphocyte serum (ALS) of immunosuppressive effect no bacterial translocation (BT) was observed in mice. The ALS treatment applied in combination with other immunosuppressive agents such as lymphotropic cytostatics as dianhydrogalactitol (30 mg/kg) or chlorpromazine (75 mg/kg) did not increase the mice drug sensitivity to used agents. According to our results, ALS can be suitable for combined application with other immunosuppressive agents as it can increase immunosuppression without side-effects such as those induced by bacterial translocation. PMID- 10389301 TI - [Components of Serratula species; screening for ecdysteroid and inorganic constituents of some Serratula plants]. AB - Ecdysteroid and inorganic components were analyzed from several plant species belonging to Caryophyllaceae family, such as in the cases of Serratula tinctoria, Serratula wolffii, Serratula coronata (20-hydroxyecdysone and inorganic components) and Jurinea mollis, Serratula gmellinii and Leuzea carthamoides (inorganic components, only). The 20-hydroxyecdysone content was determined using thin-layer chromatography after a simple clean up that had been performed by solid-phase extraction. Inorganic constitutents were determined using either ICP or flame photometry. Vegetation dependence of both 20-hydroxyecdysone and inorganic elements was studied. Serratula coronata shows remarkable high 20 hydroxyecdysone (2.3%, in April) and the studied Serratula plants gave a minimum of 20-hydroxyecdysone in June. Favorable period for harvesting is suggested as July and August, through the blossoming of these plants. Potassium and 20 hydroxyecdysone content gave a similar tendency considering their vegetation dependence, while the magnesium content moved toward opposite direction. The calcium contents of Serratula tinctoria, Serratula wolffii and Serratula coronata were found between 2.2% and 3.8%, which values are high relative to the other medicinal plants. At the same time, the Cu content of the ecdysteroid producing (and screened) Caryophyllaceae plants is low. The Fe, Mn and Mg contents of Serratula coronata are high, even higher than that of the Leuzea carthamoides. Our results have suggested the importance of analysis and control of inorganic constituents of crude plant extracts used for medicinal and recreational purposes. PMID- 10389302 TI - [Headspace solid phase micro-extraction method optimization for residual solvent analysis]. AB - A systematic headspace SPME method optimization is described for the residual solvent analysis using a polidimethyl-siloxane/divinylbenzene (PDMS/DVB) fiber. The first step was the chromatographic system optimization in which a narrow bore capillary column with a thick film (1 micron) of stationary phase, low starting column temperature (30 degrees C) and a narrow bore injector liner (2 mm I.D.) were used. It was found that for the investigated components a desorption temperature of minimum 150 degrees C should be used. The second step was the extraction optimization. The optimum equilibration time for all components was 30 min. It was found that the sample headspace volume has an important effect on method sensitivity and precision. At low headspace volumes (less than 1/3 of vial volume) sensitivity improves but at the same time precision worsen. The optimum headspace volume was found to be 4.6 ml. The total organic content does also have an important effect on method sensitivity and precision. At low organic content sensitivity increases but precision drops significantly. Over 1% organic content in the sample the system becomes unstable due to stationary phase swelling by the organic components. The optimum range for total organic content was found to be between 0.1% and 1%. The added NaCl quantity does increase the extraction yield. The optimum salt quantity to be added was 2 g NaCl in 5 ml sample. The last step was the desorption optimization. The influence of injector temperature and injection depth on desorption were investigated and it was found that it does not have an important effect on desorption yield. The optimum desorption temperature was 220 degrees C and the optimum injection depth was 2 cm into the injector. Among the investigated fibers the best detection limits and chromatographic behavior were given by the PDMS/DVB fiber. The measured detection limits were between 10 and 100 pg/ml and the RSD data were between 1-3%. PMID- 10389304 TI - [Antiretroviral treatment in 1999: from uncertainty to hopefulness]. PMID- 10389303 TI - [Characterization of drug, narcotic and psychotropic drug chirality by statistical methods]. AB - The percentage of chiral entities among drug, narcotic drug and psychotropic compounds is steadily increasing. Receptors of the human body recognize the enantiomeric forms of constitutionally identical compounds as entirely different chemical agents. Based upon these facts, this paper reports the percentage of chiral compounds in the various pharmacological classes, and related data. Pertinent terms, such as eutomer, distomer, eudismic index, eudismic affinity quotient are defined. Differences in biological activity between eutomers and distomers are exemplified. The pharmacological classes and subclasses of highest chirality, and the "most chiral" active principles are shown. Some puzzling observations on pharmacological behaviour of stereoisomers are highlighted. The necessity of "racemate switch" in the pharmaceutical industry, and the significance of stereo-specific interactions between the drug, narcotic drug and psychotropic ligands, and complementary, "pocket" moieties of the human body are emphasized. Some features of enantiopharmacology, a fledgling science in the interface of stereochemistry and traditional pharmacology are introduced. The statistical treatment of asymmetric compounds in pharmacological classes and subclasses shows that presently, the percentage of chirality in drug categories is more characteristic of the origin of the compound than its target molecule. PMID- 10389305 TI - [Clinical and immuno-virologic efficacy of the expanded access use of protease inhibitors for HIV-1 advanced disease]. AB - OBJECTIVE: To compare the efficacy and tolerance of additive therapy with protease inhibitors (PI) in patients with advanced HIV infection previously treated with retro-transcriptase inhibitors (RTI). METHODS: Eighty patients with prior antiretroviral therapy with RTI (zidovudine, ddI or ddc) for more than 6 months were included. Fifteen patients received indinavir, 42 ritonavir and 23 saquinavir. Data were collected at 4, 12 and 24 weeks and included clinical events, tolerability, plasma HIV-1 RNA viral load and CD4+ cell counts. Virologic response was defined if a viral load reduction > 1 log was achieved. RESULTS: Virological response was observed in 45 patients (56.5%) at 4 weeks and was maintained in most of them at 24 weeks. Viral load below limit of detection was achieved in 11 (15%) patients at 12 weeks. Adverse effects were not uncommon, specially with ritonavir, and 10 patients (12.5%) discontinued treatment. Indinavir was the most efficient drug and statistical differences in decreasing viral load were reached in pairwaise comparison with saquinavir at any time and with ritonavir at 12 and 24 weeks. CD4+ cell counts increased with all three drugs parallel with the decrease of viral load. Four patients died and 12 had opportunistic infections. Proportion of patients without infections in the follow up was associated with virological response over treatment (p < 0.01). CONCLUSIONS: The additive therapy with PI in advanced HIV patients can achieve a sustained reduction of viral load and a persistent recovery of CD4+ cell counts with clinical benefits. Within the limits of this study, indinavir seems more interesting in this group of patients in terms of probability pursuit of treatment because of better efficiency and fewer adverse effects. PMID- 10389306 TI - [Survival of patients with AIDS treated with protease inhibitors]. AB - OBJECTIVE: Make a volarization of the effect on survival of the protease inhibitors used on patients with established-AIDS. METHODS: Retrospective study on patients diagnosed of AIDS between January 1989 and March 1998. The main objective is the time between diagnosis and dead. It is compared the survival curve of the patients on treatment with protease inhibitors (PI) with the ones without them. We use the methods of Kaplan-Meier and log-rank. RESULTS: We analyzed 99 patients diagnosed of AIDS. Fifteen were treated with PI in combination and eighty-four with regimens without them. The number of CD4 and the age at the beginning of the study, the type of transmission and the gender were similar at the two groups. The median survival of the treated with PI was 47 (4 months (CI 95%: 39-55), and the one of the no treated as 26(3 months (CI 95%: 20 32) (p = 0.0027) CONCLUSIONS: The treatment with Protease inhibitors plus other antiviral medications is associated with a survival prolongation in patients with AIDS. PMID- 10389307 TI - [Serum and pulmonary angiotensin converting enzyme as a marker of acute lung injury in an experimental model of adult respiratory distress syndrome]. AB - OBJECTIVE: To know if the determination of the angiotensin converting enzyme in serum (SACE) and lung (LACE) may be useful as a marker of acute lung injury (ALI) in the adult respiratory distress syndrome (ARDS). METHOD: By reproducing in a experimental model of ALI with oleic acid in dogs which simulate the early stage of ARDS, we have correlated the pathologic and analytical changes observed with the results of the determinations of SACE and LACE. RESULTS: We have found sequential pulmonary lesions (congestion, edema, hemorrhage polynuclear infiltration and thrombosis) and biological alteration (hypoxemia, pulmonary hypertension, early leukopenia and final leukocytosis, thrombopenia and hypofibrinogenemia) that reproduce the typical changes of ARDS, together with the decrease of SACE--slow and progressive--and LACE--abrupt in the onset and maintained during the experiment--. CONCLUSIONS: The LACE is a good marker of the beginning of the lesion because its decrease coincides with the first pathological changes (congestion) and with the hypoxemia, pulmonary hypertension and leukopenia, maintained without changes, during the whole experiment. On the other hand, the SACE corresponds as an inespecifical reactant, marker of acute inflammation and loss of pulmonary endothelium, because its progressive decrease evolutioned with the pathological lesions and the analytical changes. In conclusion, the sequential determination of SACE has a prognostic and evolutive value in comparison with the LACE, which has a diagnostic value from the beginning of the experiment of ALI and maintained throughout. PMID- 10389308 TI - [Urinary tract infection and antibiotic sensitivity in the south of Albacete, Spain]. AB - OBJECTIVES: Describing the bacteriological map of the urinary tract infections (UTI) in our area including intra and extrahospitalized patients during 1997. MATERIAL AND METHODS: Descriptive-retrospective study, 12,937 urocultives were carried out in our laboratory: 847 from hospitalized patients and 12,090 from 15 Health Centres. RESULTS: 1,527 positive cultures (12.6%) were obteined from Health Centre patients and 215 (25.3%) from hospitalized patients. Bacterial distribution was similar between them, except enterocci and negative coagulase estafilococci which were more frequent in hospitalized patients. We have observed a high prevalence of Enterobacter sp. (4%) and Pseudomonas sp. (4%) which disagrees with other studies. E.coli (presents in 69% of the positive urocultives) was widespread resistant to quinolones, pipemidic acid and nitrofuration, while it had high sensitivity to fosfomicin, cephuroxim and amoxicilin-clavulanic acid. CONCLUSIONS: The level of resistance to the quinolones is considerable in our area, so clinicians should give them up in empirical treatment and use fosfomicine, cephuroxime or amoxicilin-clavulanic acid. PMID- 10389309 TI - [Impact of gender and influence of tobacco on the etiology of patients with hemoptysis]. AB - OBJECTIVE: To evaluate the importance of the sex and the influence of tobacco on the etiology of patients with hemoptysis. METHODS: We analyzed and compared the etiology of 394 consecutive patients with hemoptysis, divided in two groups: I (males) and II (females), and stratified in relation to tobacco. RESULTS: 305 males (77.4%) and 89 females (p < 0.0001). Lung cancer was more frequent in both groups (26.9%), especially squamous cell carcinoma, but nonmalignant etiologies were more common when we considered all globally (73.1%), including chronic obstructive lung disease (COPD) (14.9%) or bronchiectasis (20.4%) as the more frequent diseases, and 14.2% of unknown etiology (5.2% if we excluded bronchitis or lung infections). Lung tuberculosis was decreased (1.5%) in relation to a increased frequency of its sequelaes or residual lesions (10%). The initial increased frequency of lung cancers or COPD in males and bronchiectasis or heart diseases in females disappeared after the stratification and elimination of the tobacco effect. CONCLUSIONS: Hemoptysis was more frequent in males and nonmalignant etiologies were more common in both groups, with similar rates as in previous literature and a low number of cases with a unknown etiology associated to a protocolized study and the increased use of diagnostic procedures. In our study, the etiology of patients with hemoptysis was sex independent, to show that the susceptibility to tobacco illness was similar in both sexes. PMID- 10389310 TI - [Multiple nodular pulmonary AA amyloidosis. A case report]. AB - Amyloidosis is a uncommon disease which affect respiratory system with well defined patterns. We describe a case of multiple nodular pulmonary amyloidosis in a 81 year old man mimicking other entities: metastatic disease, tuberculosis and other granulomatose diseases. In our patient amyloide substance was type AA, being type AL the most frequent one in nodular pulmonary amyloidosis. PMID- 10389312 TI - [Solitary hepatic mass as initial manifestation of non-hodgkin lymphoma]. AB - A 55 years-old woman was admitted to the hospital because fifteen days of malaise, right upper quadrant pain and fever. Physical exam revealed hepato splenomaly and radiologic evaluation (abdominal ultrasonography and CT) showed a solid hepatic mass with several retroperitoneal lymphadenopathies. Through an hepatic fine needle aspiration biopsy and a bone marrow biopsy, the diagnosis of a high grade non-Hodgkin lymphoma was made. The histopathological study of a retroperitoneal lymphadenopathy surgical biopsy got the definitive diagnosis. This form of liver involvement (big solid hepatic mass) by lymphoma is not usual and when it is found, it's necessary to make a differential diagnosis with primary hepatic tumors, hepatic metastases and primary lymphoma of the liver. PMID- 10389311 TI - [A case of pneumonia by cytomegalovirus in a patient with acute myeloid leukemia]. AB - The objective of this clinical case is to suggest that Cytomegalovirus (CMV) should be taken into consideration when dealing with infections produced by opportunistic microorganisms in immunocompromised patients who have fever of unknown origin and especially when accompanied by symptoms and signs of interstitial pneumonia. In the case we report of CMV active infection in a patient with acute myeloid leukemia, the anti-CMV specific treatment (ganciclovir) was iniciated following a positive polymerase chain reaction (PCR) result for CMV. The patient initially experienced an improvement but later worsened and expired. At present there are sensitive and specific techniques such as PCR which make it possible to start specific treatment as soon as the diagnosis of CMV active infection is made. PMID- 10389313 TI - [Ischemic colitis associated with antithrombin deficiency]. AB - Colonic ischemia is one of the more common disorders of the colon in elderly people; the ischemic colitis can affect young people too. The splenic flexure, the descending colon and the sigmoid are the parts most commonly involved. Only in some cases it is possible to identify the specific cause of colonic ischemia. We report a case of ischemic colitis associated with antithrombin III deficiency. PMID- 10389314 TI - [Local thrombolysis in renal artery embolism]. AB - Renal artery embolism is an infrequent entity that occurs in patients with underlying cardiac diseases. Diagnosis is usually difficult unless the index of suspicion is high. Local thrombolysis with low-dose fibrinolytic agents is an useful therapeutic intervention. We present 2 cases of renal artery embolism treated with intra-arterial urokinase and review clinical features and therapeutic options. PMID- 10389315 TI - [Body composition and constitution: constitutional syndrome (second of two parts]. PMID- 10389316 TI - [Superior vena cava syndrome caused by mediastinal lymphoma in a patient with systemic lupus erythematosus]. PMID- 10389317 TI - [Occupational lung diseases in Salamanca: 1990-1996]. PMID- 10389318 TI - [Mounier-Kuhn syndrome]. PMID- 10389319 TI - [Primary psoas abscess and epidural abscess by Staphylococcus aureus]. PMID- 10389320 TI - [IgA deficiency, cryoglobulinemia and carpal tunnel syndrome]. PMID- 10389321 TI - [Aortic endocarditis as initial manifestation of Brucella infection]. PMID- 10389322 TI - [Deep venous thrombosis of inferior extremity in a patient with AIDS and anticardiolipin antibodies]. PMID- 10389323 TI - [Thrombocytopenia associated with rifampicin]. PMID- 10389324 TI - [Outpatient gynecologic surgery in public hospitals]. PMID- 10389325 TI - [Abdominal colposacroplexy for the treatment of vaginal vault prolapse with or without urinary stress incontinence]. AB - OBJECTIVE: The aim of this study was to assess the factors of success in abdominal colposacropexy (CSP) procedures. PATIENTS AND METHODS: We performed 271 consecutive CSP between 1986 and 1997 (mean age: 48.8 years +/- 11.1). We reviewed 217 patients (80.1%). Mean duration of follow-up was 5.5 years (1-136 months). We performed: 18 CSP with Goretex mesh, 3 with resorbable mesh and 196 with Mersilene; 179 CSP with posterior colporraphy and 38 without; 208 CSP with culdoplasty (Moschowitz's procedure) and 9 without; 182 CSP with anterior and posterior meshes, 26 with posterior mesh only and 9 with anterior mesh only. RESULTS: 97.7% (212/217) of patients were cured for prolapse. 58% (125/217) had urinary stress incontinence totally cured and 82% (178/217) had urinary stress incontinence improved. Rejected grafts were 16.7% (3/18) with Goretex mesh and 1.1% with Mersilene mesh (p = 0.004). Recurrent prolapses were 1.1% (2/196) with CSP with posterior colporrhaphy and 7.9% (3/38) in CSP without (p = 0.009; OR = 0.14, CI = 0.02-0.86); 4/208 with CSP with culdoplasty and 1/9 with CSP without (p = 0.04; OR = 0.17, CI = 0.02-1.58). Recurrent stress incontinence was observed in 4/9 cases with CSP with anterior mesh only and 28/182 with CSP with anterior and posterior meshes (p = 0.03; OR = 0.34, CI = 0.12-0.97). CONCLUSION: CSP must use anterior and posterior Mersilene mesh. The CSP must be systematically combined with posterior colporraphy and culdoplasty (Moschcowitz's procedure). PMID- 10389326 TI - [Surgical treatment of pancreatic enzyme serous effusions]. AB - Enzymatic seritis is a rare complication of chronic pancreatitis. Thirty-four cases are analyzed from a series of 200 cases of operated chronic pancreatitis: 22 pleural effusions, 5 ascites and 7 combined effusions. The pancreatic leak was demonstrated preoperatively in 40% of cases. The leak originated from erosion of a pancreatic duct in 7 cases and leaking pseudocyst in 27 cases. All patients were operated: internal drainage (22 cases), left splenopancreatectomy (7 cases) and external drainage (5 cases). Postoperative mortality was 9% (n = 3); postoperative morbidity was 15% (n = 5). The effusion did not recur in any of the survivors, but repeat surgery for chronic pancreatitis complications was necessary in 7 patients (20.5%). CONCLUSION: after failure of medical treatment, the treatment of large serous effusions is surgical: internal drainage or pancreatic resection when the lesion is located in the tail of the pancreas. PMID- 10389328 TI - [Outpatient management, patient comfort and satisfaction of 100 consecutive inguinal hernias treated by Shouldice procedures with steel wire under local anesthesia]. AB - This study evaluates postoperative patient comfort and satisfaction of 100 unilateral inguinal hernia repairs performed by Shouldice procedure with steel wire and local anesthesia. The day surgery management of 78% of patients, the low analgesic consumption, the resumption of normal activity after one week and a satisfaction index of nearly 100% are the main results of this study. The quality of these results is another argument confirming this technique as the "Gold Standard" of hernia repair. PMID- 10389327 TI - [Laparoscopic cure of gastroesophageal reflux disease. Results of a multicenter trial]. AB - OBJECTIVES: The aim of our trial was to assess preoperative assessment and short term results of laparoscopic cure of gastroesophageal disease performed by digestive surgeons not specialized in this disease. METHODS: 335 consecutive cures were performed by 11 surgeons. RESULTS: 29% of patients had pH-metry, 52% had manometry. The procedure was: Nissen-Rossetti in 66% of cases, Toupet in 17%, and Nissen in 16% of cases. The conversion rate was 13%. The intraoperative complication rate was 3.28% and 2.08% of patients had a reoperation. One patient died. 5% of patients had dysphagia more than three months after the operation. CONCLUSION: Our results show an absence of consensus concerning preoperative assessment. They suggest that careful intraoperative management, with a high conversion rate, reflecting our initial experience, allows similar results to those of teams specialized in laparoscopic surgery and better than laparotomy series. PMID- 10389330 TI - [Pancreatectomy and diabetes]. AB - Post total pancreatectomy diabetes is a clearly defined form of unstable diabetes, requiring low doses of insulin, with frequent and severe hypoglycemic events. This is due to both deficiency of pancreatic glucagon, hormone of primary importance for hepatic gluconeogenesis and glycogenolysis, and exocrine failure. The management of this form of diabetes is difficult, involving exact correction of malabsorption and low doses of insulin. Whenever possible, partial pancreatectomy should therefore to be preferred. After partial pancreatectomy, the likelihood of diabetes depends on the volume of the remaining pancreas, the type of resection and above all the preexisting pancreatic status. Prevention of postoperative hyperglycemia could minimize the risk of long-term diabetes. Pancreatic cancer is a particular case: the onset of diabetes could be a manifestation of occult pancreatic cancer and glucose metabolism may improve after tumour excision with preservation of some pancreatic tissue. PMID- 10389329 TI - [General review of the value of hepatic MRI in the diagnosis and preoperative staging of liver metastasis from colon and rectum cancer]. AB - Hepatic resection of secondary liver neoplasms is currently the only potentially curative therapy for patients with primary colorectal carcinoma. Long-term survival is closely related to stage, regardless of the number, size and distribution of liver lesions. Preoperative detection of liver metastases is crucial in patient staging and imaging techniques must be as accurate as possible to evaluate whether hepatic resection can be performed. Among the various strategies applied to the preoperative detection of liver metastases, CT with intra-arterial portography (CTAP) has been found to be the most sensitive technique. It is an invasive evaluation with a reported sensitivity ranging from 81 to 94% but with 6 to 15% of false-positive results. Recent studies demonstrate that MRI with superparamagnetic iron oxide has an equivalent sensitivity and specificity to CTAP. This paper reviews the technique of MRI, evaluates its sensitivity and specificity, and presents the advantages and draw backs of the technique. PMID- 10389331 TI - [Can intestinal transplantation constitute treatment for intestinal failure?]. AB - The management of patients with intestinal failure has benefited from progress in parenteral nutrition (PN), especially home-based parental nutrition. Intestinal transplantation is now possible and in some conditions, constitutes the logical treatment option. Since 1985, more than 300 small-bowel grafts have been performed, involving the isolated small bowel with or without the colon (45%), the liver + small bowel (40%) or several organs (15%). 2/3 of recipients were under 20 years of age, and indications were short-bowel syndrome (64%), severe intractable diarrhea (13%), abdominal cancer (13%), or chronic intestinal pseudo obstruction syndrome (8%). 51% of patients survived > 2 years after the graft. Patient and graft survival depends on the type of immunosuppression, i.e. Cyclosporine or FK 506. The results must be interpreted carefully as they represent the first experience in numerous centers using different immuno suppressive protocols, without any randomization. The results from the largest of these centers more closely reflect the current situation and may exceed a 70% 2 year survival rate. Functional grafts lead to gastrointestinal autonomy (weaning of PN) while maintaining satisfactory nutritional status and normal growth in childhood. Intestinal transplantation is theoretically indicated for all patients permanently or persistently dependent on PN. However, as PN is generally well tolerated, even for long periods, each indication for transplantation must be carefully weighed up in terms of the iatrogenic risk and quality of life. When PN has reached its limits, especially those associated with vascular, infectious, hepatic or metabolic complications, intestinal transplantation must be undertaken. Transplantation of the small bowel alone remains the first option, as combined liver-small bowel grafting is only indicated in case of life-threatening progressive cirrhogenic liver disease. PMID- 10389332 TI - [Primary gastric lymphoma: towards earlier medical management. Adult Lymphoma Study Group]. PMID- 10389333 TI - [A pioneering anesthesiologist, the surgeon Theodore Tuffier]. AB - Theodore Tuffier (1857-1929) was a great surgeon. A part of his interest non well know and reported in this paper was fundamental and technical research in anesthesia. PMID- 10389334 TI - [An unusual cause of acute pancreatitis]. PMID- 10389335 TI - [Colonic intussusception in adults: a case report]. PMID- 10389336 TI - [Mucocele on bypassed esophagus: two operated cases]. PMID- 10389337 TI - [Sigmoido-ureteric fistula. Report of an exceptional case]. PMID- 10389338 TI - Let's focus on the positive. PMID- 10389339 TI - Psychiatric nurse practitioner versus clinical nurse specialist: moving from debate to action on the future of advanced psychiatric nursing. AB - The psychiatric nursing literature recently has included a proliferation of discussions regarding the nature and direction of change impacting the future of advanced practice psychiatric nursing. The debate has focused most commonly on the role of the clinical nurse specialist versus the role of the nurse practitioner. The debate has produced little in the way of outcomes other than an entrenchment of positions. The stalemate in psychiatric nursing is producing a slow but steady surrender of the boundaries of psychiatric nursing to other fields of nursing. Although advanced practice psychiatric nurses disagree on what to become and what to be called, people with conditions such as depression, anxiety disorders, and other psychiatric disorders are being treated increasingly by family nurse practitioners. The time for debate has ended. Unless consensus regarding what constitutes the domain of psychiatric nursing is reached soon, the discussions will be moot because few clients will remain to be treated. This article began as a discussion between colleagues. The two authors teach at a regional state university, but they share diverse opinions regarding the substance and nature of advanced practice psychiatric nursing. These diverse views led to discussions that have implications not only for faculty practice, but for curricular design, and for decisions regarding how to best educate future nurses. The discussion developed into a presentation at the 20th Southeast Conference of Clinical Nurse Specialists. It was presented as a point counterpoint discussion regarding this debate; one author advocated the perspective of traditional clinical nurse specialist and one advocated the perspective of a psychiatric nurse practitioner role. We conclude with a projected model of a merged role, with delineation of traditional clinical nurse specialist and nurse practitioner that must be blended for the new role. PMID- 10389340 TI - Ambient sound levels in a state psychiatric hospital. AB - High environmental and psychosocial stress contribute to the onset and relapse of major psychiatric disorders. High sound levels in general hospitals are common and may be indirectly associated with negative physical effects because of increased physiological stress on the body. Excessive sound also interferes with cognitive functioning, especially affecting prefrontal cortical processes, but no information about sound levels in psychiatric hospitals was available. This study critically examines literature on sound stress and reports findings from an exploratory study of sound levels in a tertiary care psychiatric hospital. An overall mean sound level of 75.68 dB was found, with peak sound levels as high as 85 to 90 dB, in the range that causes hearing loss. These levels, higher than sound levels on medical, surgical, and intensive care units, suggest the need for more attention to the effect that environmental sound has on the behavior of patients hospitalized with acute psychiatric symptoms. PMID- 10389341 TI - Psychosocial issues associated with increased breast and ovarian cancer risk: findings from focus groups. AB - The advent of genetic testing for breast and ovarian cancer susceptibility has raised concurrent psychosocial issues. Within the context of limited health care resources, the ability to identify a subgroup of women who are at increased genetic risk for breast and ovarian cancers and who are also vulnerable to significant psychosocial sequelae is critical. Use of a focus group methodology substantiated the notion that there are factors that may predispose certain women who are at increased risk for developing breast and ovarian cancer to sustained psychosocial problems. In this era of rapid scientific strides coupled with efforts to contain health care costs, it is imperative that screening instruments be developed that can identify women who are at risk for significant psychosocial sequelae so that interventions can be instituted in a timely fashion. PMID- 10389342 TI - The Heideggerian view of person: a perspective conducive to the therapeutic encounter. AB - The artistry of the therapist finds its origins in the therapist's authentic presencing of self and in the philosophical assumptions that constitute the therapist's perception of human beings. Based in existential phenomenology, Heidegger's view of person nurtures a conceptualization of individuals complimentary to the art of person-centered psychotherapy. Heideggerian hermeneutic phenomenology encourages ontological self-reflection and potentiates meaningful appreciation of one's "being-in-world." This study describes Heidegger's view of person as one that offers therapists and their patients an advantageous clinical orientation and an anesthetically comfortable realm from which the art of the therapeutic encounter can originate, be nurtured, and grow infinitely, thereby supporting patients' self-transcendence, self-love, and well being. PMID- 10389343 TI - The use of critical pathways in caring for schizophrenic patients in a mental hospital. AB - To provide quality health care and at the same time, to control cost, literature suggests that using critical pathways as a tool can enhance resource management, increase collaborative practice, and benefit patient care. This study describes the processes of developing a critical pathway in caring for schizophrenic patients in a mental hospital in Hong Kong. The perceived benefits and difficulties in using the critical pathway are discussed from a nursing perspective. Nurses believed that the use of critical pathways could improve the coordination and effectiveness of care. Also, nurses' autonomy and professional status improved. However, inadequate knowledge and resistance from other disciplines were barriers to the implementation. Recommendations are given to overcome the barriers. PMID- 10389344 TI - Perioperative cardiac risk assessment for noncardiac surgery. AB - Patients with coronary artery disease who undergo noncardiac surgery are at risk of cardiac complications. It is, therefore, imperative to assess the risk of noncardiac surgery in these patients in a variety of operative settings. We propose a simplified approach at perioperative risk assessment that ascertains risks based on the need and urgency of surgery, and the type of surgery. Furthermore, we suggest guidelines for perioperative risk assessment based on patient's history and physical exam, their functional capacity and prior procedures. Based on these clinical data, we identify patients into three categories for a cardiac event during noncardiac surgery: high, intermediate and low risk groups. We recommend that all urgent surgeries should be carried out with attempts made at proper perioperative management to reduce the risk of cardiac events. Further, low or intermediate risk patients undergoing minor surgery should be operated without any further cardiac evaluation. Intermediate risk patients, who need an intermediate or major surgery, should undergo a stress test to evaluate for ischemia. If there is evidence of a large ischemic burden and those patients at high risk should undergo cardiac catheterization and appropriate intervention. We also suggest methods of modifying risk in the perioperative period for all patients to reduce perioperative cardiac events with proper monitoring and use of medications including beta blockers. This article provides a detailed review on the current literature supporting the stepwise approach proposed by us. PMID- 10389345 TI - Biochemical assessment of myocardial damage with new diagnostic tools. AB - Knowledge of the pathophysiology of acute coronary syndrome has advanced in parallel with awareness of the significant limitations inherent in clinical and electrocardiographic assessment. In the meantime, biochemical assays, long established as a reliable means of detecting World Health Organization-defined myocardial infarction, have improved significantly. Prior of the late 1980s, diagnostic testing for so-called "cardiac" enzymes was not enough sensitive and cardiac-specific. New immunochemical techniques for measuring the mass rather than the activity of an enzyme permit the detection of smaller amounts of the protein markers, and therefore earlier diagnosis is possible. In addition, new laboratory techniques now allow rapid measurement of a wide variety of markers of myocardial cell death. Several serum proteins have recently been introduced and evaluated to determine sensitivity and specificity for detection of acute myocardial damage, including creatine kinase-MB mass measurements, myoglobin, cardiac troponin I, and cardiac troponin T. Cardiac troponin measurement is probably the most clinically effective test, also offering prognostic information to the clinician unavailable by other methods. The diagnostic superiority of these proteins has been established by clinical studies and rigorous statistical comparison with other markers. Furthermore, there is interesting evidence that minor myocardial cell injury revealed by these markers is strictly associated with a high risk of developing major cardiac events during follow-up. This means that these markers should replace existing traditional strategies to guide patient management. Obviously, there will also be financial benefits in terms of reduction in patient episode cost and improvement in decision-making. PMID- 10389346 TI - Treatment of hypertension in the elderly. AB - Several trials performed in elderly patients have demonstrated that antihypertensive drugs are effective in both systo-diastolic and isolated systolic hypertension, reducing the incidence of fatal and nonfatal cardiovascular events. However because of technical and design problems, the studies carried out to date have involved highly selected patients, almost always without any target organ damage, independent, cognitively normal and with low comorbidity. Therefore, trial results may be transferred to clinical practice only with some caution. Therapeutic behavior could be different in the presence of diseases associated with hypertension: a) in case of associated specific cardiovascular complications and/or diseases, such as diabetes or dyslipidemia, which could increase cardiovascular risk, treatment must be more aggressive; b) in case of associated diseases with fatal prognosis, treatment is aimed at preventing hypertensive emergencies; c) in case of associated diseases, which are not life-threatening but require chronic pharmacological intervention, drug interaction must be carefully considered. Finally, sudden and significant blood pressure drops due both to overdosage of antihypertensive drugs and/or to intercurrent illnesses must be prevented, because the reduction of blood flow may induce severe target organ ischemia. PMID- 10389347 TI - Genetic basis of cerebrovascular accidents associated with hypertension. AB - Among hypertension-associated cardiovascular diseases, stroke represents one of the most common disorders. In fact, it significantly affects mortality and morbidity rates of all industrialized countries. Only recently, stroke has been considered as a complex trait and not as a mere consequence of hypertension. Indeed, it appears to be the result of an interaction among several genetic and environmental factors. The identification of the genetic determinants of stroke is a difficult task in humans, due to the genetic heterogeneity of human populations and the confounding presence of other risk factors. Thus, an experimental approach, through the use of a highly inbred animal model for stroke, offers a valuable alternative and additional support for a genetic dissection of cerebrovascular disease. In fact, the genetic analysis of stroke in the animal model of the stroke-prone spontaneously hypertensive rat provided clear evidence that stroke is a genetically determined complex trait, and that factors such as blood pressure and diet only play a permissive role. Finally, by using the experimental approach, we established that the gene encoding atrial natriuretic peptide, significantly linked to cerebrovascular disease in rats, is a genetic determinant of stroke in humans. The identification of the genetic basis of stroke represents an important step towards the institution of targeted preventive and therapeutic approaches to reduce the risks of cerebrovascular accidents. This article reviews the background, the experimental approach and the outcome of a strategy based on the use of the stroke-prone spontaneously hypertensive rat model, which aims at identifying the genetic basis of stroke. PMID- 10389348 TI - Should ablation of atrial flutter be discouraged in patients with documented atrial fibrillation? AB - Atrial fibrillation and atrial flutter often coexist in the same patient. The purpose of this article is to provide an analysis of the mechanisms underlying the transformation from atrial fibrillation into atrial flutter and to investigate the long-term clinical benefits following ablation of atrial flutter in relation to recurrences of atrial fibrillation. Experimental studies in the human atrium demonstrated that in most instances atrial fibrillation is a triggering rhythm for atrial flutter. However, a review of the most recent studies shows a low percentage of recurrence of atrial fibrillation after successful catheter ablation for atrial flutter. The risk factors for this recurrence are the presence of structural heart disease, increased left atrial dimension and volume, a previous history of atrial fibrillation, and the failure of multiple antiarrhythmic drugs, inducibility of atrial fibrillation by a standard programmed electrical stimulation protocol after catheter ablation. These data, together with the high success rate of catheter ablation for atrial flutter, suggest to perform radiofrequency catheter ablation for atrial flutter in patients with documented atrial fibrillation. PMID- 10389349 TI - [Effects of chronic subclinical hyperthyroidism from levothyroxine on cardiac morphology and function]. AB - BACKGROUND: Thyroid hormones greatly affect the cardiovascular system. Although the effects of overt hyperthyroidism on the cardiovascular system have been diffusely studied, only in the last years the effects of subclinical hyperthyroidism on the heart have been investigated. Subclinical hyperthyroidism is a symptomatic or asymptomatic condition with an absent response of thyrotropin (TSH) to thyrotropin-releasing hormone in the presence of normal serum levels of thyroid hormones for the general population, though supraoptimal for the individual. The more frequent causes of endogenous subclinical hyperthyroidism are multinodular goiter, toxic, adenoma and Graves's disease, whereas the exogenous causes are induced by levothyroxine (LT4) therapy used to suppress TSH in patients with nontoxic goiter and differentiated thyroid cancer. This paper reports our experience derived from the study of 60 patients with subclinical hyperthyroidism due to TSH-suppressive therapy with LT4 compared to normal subjects. METHODS: Patients (9 males and 51 females, mean age 39 +/- 10 years) were studied by complete Doppler echocardiography, standard and 24 hour ECG Holter monitoring, exercise test with cycloergometer, and radionuclide ventriculography at rest and during fixed workload (75 W). RESULTS: Holter monitoring showed a significant increase in mean 24 hour heart rate (80 +/- 10 vs 70 +/- 9 b/min, p < 0.001) and supraventricular arrhythmias (42 vs 12 patients, p < 0.003). Echocardiography showed an increase in left ventricular mass index (94 +/- 13 vs 80 +/- 18 g/m2, p < 0.001) due to increased septal and posterior wall thickness. At rest, echocardiographic indices of systolic function (fractional shortening and mean corrected velocity of circumferential fiber shortening) were higher in patients than in controls (fractional shortening 40 +/- 6 vs 34 +/- 4%, p < 0.001; mean corrected velocity of circumferential fiber shortening 1.23 +/- 0.17 vs 1.05 +/- 0.14 circ/s, p < 0.001), while the Doppler indices of diastolic function were significantly impaired as documented by the reduced E/A ratio (1.18 +/- 0.3 vs 1.8 +/- 0.5, p < 0.001) and the prolonged isovolumic relaxation time (94 +/- 13 vs 78 +/- 12 ms, p < 0.001). Exercise tolerance was also significantly impaired in patients with subclinical hyperthyroidism: maximal exercise time (6.4 +/- 0.7 vs 9.4 +/- 1.4 min, p < 0.001) and peak workload (81 +/- 11 vs 121 +/- 17 W, p < 0.001) were significantly reduced and radionuclide ventriculography showed a decrease in ejection fraction during exercise (from 62 +/- 7 to 53 +/- 8%, p < 0.002). CONCLUSIONS: Persistent subclinical hyperthyroidism by TSH-suppressive doses of LT4 significantly affects heart morphology and function. Thus, we suggest that a complete suppression of TSH must be recommended only in patients with differentiated thyroid cancer, while in patients with begin thyroid disease it could be sufficient to maintain subnormal TSH levels. PMID- 10389350 TI - The impact of second harmonic imaging on stress echocardiography reading. AB - BACKGROUND: Noncontrast harmonic significantly improves overall image quality in echocardiography. The aim of this study was to assess whether harmonic imaging, without contrast, impacts on interobserver variability and diagnostic accuracy of "beginners" in stress echocardiography. METHODS: Images at rest and peak stress were obtained in digitized format in 15 consecutive patients (10 males, 5 females, mean age 66 +/- 9 years) and analyzed by 5 inexperienced observers (stress echo beginners) who were blinded to the imaging modality (standard versus harmonic imaging). Each observer graded the image quality as 1 = uninterpretable up to 5 = excellent, in a total of 240 segments. RESULTS: The mean image quality per segment was 2.9 for conventional technology and increased up to 3.6 for harmonic imaging (p < 0.001). The interobserver agreement (> or = 4 out of 5 readers) rose from 46 to 60%. The percentage of uninterpretable segments was 7.5% at rest and 8.9% at peak stress of conventional imaging, and decreased to 4.6 and 6.5%, respectively (p < 0.05) by second harmonic technology. The unanimous reading of two additional independent expert observers was arbitrarily assumed to be the "gold standard" to verify the accuracy of reading of the 5 beginners. The 5 beginners showed poor diagnostic accuracy with fundamental imaging (73%), and they did not improve with second harmonic imaging (70%, NS vs fundamental). CONCLUSIONS: Noncontrast second harmonic imaging improves image quality over conventional imaging. The improved quality of the image deflated interobserver variability but did not determine per se an improvement in the diagnostic accuracy of nonexperienced readers. PMID- 10389351 TI - Echocardiography with tissue harmonic imaging: basic principles. PMID- 10389352 TI - New technologies in two-dimensional echocardiography: marketing gadget or clinical tool? PMID- 10389353 TI - Predictors of tilting test negativization during beta-blocker treatment in patients with neurocardiogenic syncope. AB - BACKGROUND: The cardioinhibitory and/or vasodepressor reflex, always preceded by adrenergic activation, has recently been accepted as the main mechanism of neuromediated syncope. The aim of this study was to verify if efficacy of beta blockers, in the treatment of neuromediated syncope, may be predicted on the basis of clinical variables and data derived from tilting test. METHODS: We retrospectively analyzed 23 patients with recurrent or traumatic unexplained syncopal episodes, with a positive tilting test, who repeated the test during beta-blocker therapy. According to the second tilting test results, patients were divided into Group 1 (19 patients) with a positive second test, and Group 2 (4 patients) with a negative second test. RESULTS: No difference was found between the two groups in age, gender, number of syncopal episodes before observation, and period (days) between the two tests. Moreover, there was no difference between the two groups in the kind of positive response to tilting test, in heart rate at tilting (minimum, maximum, delta and slope of increase), and in the minimum, maximum and slope of heart rate increase before syncope. Delta heart rate before syncope was 15 b/min in Group 1, and 28 b/min in Group 2 (p = 0.011). Taking a discriminant value of 20 b/min, 100% sensitivity and 68% specificity were found. CONCLUSIONS: Heart rate increase before syncope identifies patients with negativization of tilting test on beta-blocker therapy. However, the clinical value of the second test for driving therapy is controversial. PMID- 10389354 TI - Cardiac variant of Fabry's disease mimicking hypertrophic cardiomyopathy. AB - A case of cardiac variant of Fabry's disease mimicking hypertrophic cardiomyopathy is reported. The diagnosis was obtained by biventricular endomyocardial biopsy showing severely hypertrophied myocardiocytes with large periodic acid-Schiff and Sudan black positive perinuclear vacuoles, shown at electromicroscopy to consist of lamellated cytoplasmic figures highly suggestive of Fabry's disease, and confirmed by diagnostic low activity of alpha galactosidase A in the peripheral lymphocytes. Invasive approach was suggested by the occurrence of a long-standing atrial fibrillation that failed to determine deterioration of cardiac function. Differential diagnosis between hypertrophic cardiomyopathy and the cardiac variant of Fabry's disease is relevant for prognostic and therapeutic implications including the perspective of an enzyme replacement therapy. PMID- 10389355 TI - Pharmacotherapy of behavioral disturbances associated with dementia. PMID- 10389356 TI - Psychogeriatrics, 1999. PMID- 10389357 TI - The Israel Journal of Psychiatry: a self-evaluation. PMID- 10389358 TI - Validation of a Hebrew selective reminding test. AB - We developed a Hebrew version for the Buschke Selective Reminding Memory Test, with three parallel forms. This Hebrew version was administered in counterbalanced order to 24 normal subjects aged 14-77 years. We studied the reliability between parallel forms, and the validity and sensitivity memory reduction in normal aging. Data were compared to American norms. The three Hebrew forms were found to be of equal difficulty, with correlation coefficients of .6 to .7 (p's < .01). Age affected the great majority of memory performances, i.e., lower performance with increasing age. Test performance was equivalent to American norms within 6%. We conclude that this Hebrew version is reliable and valid, and can be used on Hebrew-speaking populations to assess memory functions. PMID- 10389359 TI - Memory versus intelligence in dementia screening--MMSE. AB - The Mini-Mental State exam is a widely used screening instrument for dementia. Recent research has suggested that errors in classification reported for this instrument may be due to premorbid levels of intelligence and education, Thirty one ambulatory patients diagnosed with Alzheimer's disease were administered a standard neuropsychological testing battery. MMSE scores and intelligence are significantly correlated. However when memory ability is partialed out, the covariance of MMSE and intellectual abilities does not add any further information to that already provided by the memory scores. PMID- 10389360 TI - How do cognitively impaired elderly patients define "testament": reliability and validity of the testament definition scale. AB - The testament definition scale (TDS) is a specifically designed six-item scale aimed at measuring the respondent's capacity to define "testament." We assessed the reliability and validity of this new short scale in 31 community-dwelling cognitively impaired elderly patients. Interrater reliability for the six items ranged from .87 to .97. The interrater reliability for the total score was .77. Significant correlations were found between the TDS score and the Mini-Mental State Examination (MMSE) and the Cambridge Cognitive Examination scores (r = .71 and .72 respectively, p = .001). Criterion validity yielded significantly different means for subjects with MMSE scores of 24-30 and 0-23: mean 3.9 and 1.6 respectively (t(20) = 4.7, p = .001). Using a cutoff point of 0-2 vs. 3+, 79% of the subjects were correctly classified as severely cognitively impaired, with only 8.3% false positives, and a positive predictive value of 94%. Thus, TDS was found both reliable and valid. This scale, however, is not synonymous with testamentary capacity. The discussion deals with the methodological limitations of this study, and highlights the practical as well as the theoretical relevance of TDS. Future studies are warranted to elucidate the relationships between TDS and existing legal requirements of testamentary capacity. PMID- 10389361 TI - Diogenes syndrome and hoarding in the elderly: case reports. AB - Presented here are two case reports of elderly persons with Diogenes syndrome (variously known as senile breakdown, social breakdown and senile squalor syndrome). Diogenes syndrome is often (but not always) characterized by a tendency to hoard excessively (syllogomania). The first patient was diagnosed as having both a schizotypal personality disorder and obessive-compulsive disorders (OCD) while the second was diagnosed as having a schizoid personality disorder. Only the former demonstrated the tendency to hoard rubbish. The Diogenes syndrome in both cases can be hypothesized to be a reaction to stress in elderly people with certain personality characteristics or as the end stage of a personality disorder. The hoarding behavior that was manifested only in the first case can probably be the result of the presence of an OCD. The authors raise the possibility that OCD may be the cause of hoarding rubbish in those cases of Diogenes syndrome in which hoarding exists and cannot be explained by psychotic disorders, dementia or any other mental disorders due to a general medical condition (GMC) or substance-related disorders. PMID- 10389363 TI - Amnestic state in a Holocaust survivor patient: psychogenic versus neurological basis. AB - Differentiation between psychogenic and organic amnesia is sometimes quite difficult. This paper focuses on the psychogenic and organic components of a complex case of amnesia rooted in remote and prolonged traumatic stress and manifested under circumstances evoking dissociated memories. The Transient Global Amnesia (TGA) of a concentration camp survivor who developed sudden amnesia during a psychiatric intake interview was clearly triggered by the pressure of repressed Holocaust memories. The importance of distinguishing between TGA and dissociative amnesia is emphasized, and the role of psychological upset as a precipitant in TGA is stressed. PMID- 10389362 TI - Hoarding: a review. AB - BACKGROUND: Hoarding is currently categorized as a symptom of both obsessive compulsive disorder (OCD) and obsessive-compulsive personality disorder. However, hoarding has also been documented in association with other psychiatric disorders, raising questions about the classification, psychobiology and treatment of these symptoms. This paper reviews the literature on hoarding. METHOD: A computerized literature search was done using the MEDLINE database. Relevant references were collated and were used to obtain additional literature on hoarding behavior. RESULTS: Although hoarding may meet DSM-IV diagnostic criteria for a compulsion, this symptom is also seen in a range of other disorders. Nevertheless, the phenomenology of hoarding remains under-researched. In addition, the psychobiology and treatment of hoarding remains relatively poorly understood, although in certain aspects there may be some overlap with the symptoms of OCD. CONCLUSIONS: Further research on the phenomenology, psychobiology and treatment of hoarding is clearly needed. A neuro-ethological view of hoarding as a spectrum symptom seen not only in OCD, but also in other disorders, may provide a useful heuristic for current clinical practice and for future empirical research. PMID- 10389364 TI - Diagnosis of Holocaust survivors and their children. AB - Survivors of the Holocaust and their children have tended not to be given formal diagnoses by their therapists. There seem to be a series of reasons: the events themselves were so terrible that it seems inappropriate to focus on the response, diagnosis implies comparing the condition with responses to other more minor traumata, the process of diagnosis is dehumanizing, the evil nature of the perpetrator is neglected, therapists feel it distances them from their patients, and it ignores the extraordinary achievements of many survivors who cope and live full lives. The DSM and its five axes are proposed as a suitable diagnostic vehicle, and Holocaust survivors with serious symptoms will tend to be diagnosed as chronic PTSD, child survivors as complex PTSD often with associated personality disorders, and second generation may well have identity problems and personality disorders. Only by using diagnoses can comparable research be carried out. PMID- 10389365 TI - Psychiatry in the Palestine Authority: legal, ethical and forensic issues. AB - The historical and political reality of life in the West Bank and Gaza has deeply influenced the development of psychiatric services, and particularly the provision of evaluation and care for Palestinian forensic patients by Palestinian psychiatrists. The difficulties are presented and how they have changed during the events of the last decades. PMID- 10389366 TI - Topical and oral agents for erectile dysfunction. AB - Innovative research in the past 2 decades has shown that the hemodynamics of penile erection involve arterial dilation, sinusoidal expansion, and venous compression. Relaxation of the intracorporeal smooth muscles (trabecular and arteriolar) seems to be the final common pathway that leads to the above events. Neuropharmacologic studies have also established nitric oxide as the principal neurotransmitter for penile erection and confirmed the importance of cyclic adenosine monophosphate and cyclic guanosine monophosphate systems in penile smooth muscle relaxation. Recent years have also witnessed dramatic changes in the therapy of erectile dysfunction. The penile prosthesis, a gold standard of therapy of the 1970s, was replaced in the 1980s by the intracavernous injection and the vacuum constriction device. In the 1990s, two revolutionary concepts in erectile dysfunction therapy were added: transurethral alprostadil and oral sildenafil. However, the tremendous publicity surrounding the recent introduction of sildenafil has also created socioeconomic and ethical dilemmas, especially with regard to insurance coverage and government regulation. Medically, many problems also surfaced when large numbers of erectile dysfunction patients overwhelmed primary care physicians who were unfamiliar with the diagnosis and treatment of erectile dysfunction. This article reviews the advances in penile physiology and the clinical usefulness of topical and oral agents. In addition, a patients' goal-directed approach to the diagnosis and treatment of erectile dysfunction is presented. PMID- 10389367 TI - Ultrastructural changes of the tooth root surface by Nd:YAG laser irradiation followed by citric acid and tetracycline. AB - The neodymium:yttrium-aluminum-garnet (Nd:YAG) laser has been used for treatment of dentinal hypersensitivity, eradicating periodontal pathogens, and facilitating calculus removal. However, Nd:YAG laser irradiation exerts potentially harmful effects on the tooth root surface. The purpose of this study was to examine the ultrastructural changes of the tooth root surface caused by Nd:YAG laser irradiation, and to determine whether chemical and mechanical preparations can correct these ultrastructural changes. Eighteen tooth specimens (3 x 3 x 0.5 mm) with healthy root surfaces were prepared and irradiated with an Nd:YAG laser at various power densities. Root surfaces were irradiated at 100 mJ at 20 pulses per second (pps) for 2 seconds followed by the application of citric acid (pH 1.2) or tetracycline solution (100 mg/mL) for 3 and 5 minutes, respectively, or ultrasonic scaling for 5 strokes of 3 seconds per stroke. As observed with low vacuum scanning electron microscopy, (Wet-SEM), Nd:YAG laser irradiation at 70 to 100 mJ, 20 pps for 2 seconds caused surface cratering, areas of porosity, pitting, fissures, and lava-like structures in an area 140 to 280 microns in diameter. Irradiation of 50 mJ, 20 pps for 2 seconds, led to only mild surface charring. No evidence of morphologic changes was found when root surfaces were irradiated with the Nd:YAG laser at 20 mJ, 20 pps for 2 seconds or at 50 mJ, 10 pps for 8 seconds. The laser-induced lava-like structures were partially detached by citric acid (pH 1.2) etching and ultrasonic scaling, but not by tetracycline (100 mg/mL). These results indicate that chemical and mechanical preparations can be used effectively in conjunction with Nd:YAG laser irradiation for root surface preparation during both nonsurgical and surgical periodontal treatments. PMID- 10389369 TI - Measurement of the quality of life during different clinical phases of breast cancer. AB - This cross-sectional study was designed to evaluate the self-rated quality of life of Taiwanese breast cancer patients at the time of diagnosis and during various phases of treatment. A total of 115 patients at different clinical stages of breast cancer completed the study. The questionnaires consisted of the Medical Outcome Survey 36-Item Short Form Health Surveys (SF-36), the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire- Cancer 30 (EORTC QLQ-C30), and questions measuring utility using visual analog scale (VAS), standard gamble (SG), and time trade-off (TTO) methods. The patients filled out the questionnaires, with interviewers' assistance as requested. Cronbach's alpha coefficients of internal consistency for the SF-36, based on the US factor structure, were 0.84 for raw scores and 0.87 and 0.88 for the physical and mental components, respectively. Cronbach's alpha for the EORTC QLQ-C30 was 0.86. The correlations between items in the SF-36 and the EORTC QLQ-C30 that examined similar dimensions were high. Significant differences were found in most dimensions of quality of life across different clinical stages. The three utility scores, however, showed no significant differences among patients in different clinical stages. Patients in the chemotherapy and recurrence phases usually had the lowest quality of life scores, while those in the follow-up phase had the highest. The results of this study demonstrate the applicability of quality of life measurements in helping health professionals identify the physical, mental, and social problems of breast cancer patients in different phases of the clinical process. PMID- 10389368 TI - Evaluation of a simplified staging system for prognosis of hepatocellular carcinoma. AB - The current TNM (tumor, nodes, metastases) staging system for human hepatocellular carcinoma (HCC) has been challenged since a new T staging system was proposed to correlate the staging group with patient outcome after curative liver resection. The new T staging system proposed T1 as no vascular invasion, small size (< or = 5 cm), and solitary tumor. T2 was defined as the presence of one of the following factors: size greater than 5 cm, vascular invasion, or multiple tumors; T3 as the presence of two of the above three factors; and T4, the presence of all three factors. A total of 323 patients undergoing curative partial hepatectomy for HCC were studied. Kaplan-Meier survival analysis was used to evaluate the postoperative outcome. The new T staging showed good correlation between the staging group and patient outcome. The 1-year disease-free survival (DFS) rate and overall survival (OS) rate were 80.0% and 87.8% for stage 1 (n = 115), 67.6% and 81.6% for stage 2 (n = 136), 40.0% and 58.0% for stage 3 (n = 58), and 21.4% and 42.8% for stage 4 (n = 14), respectively. The 3-year DFS rate and OS rate were 61.0% and 64.5% for stage 1, 37.8% and 50.7% for stage 2, 21.4% and 29.8% for stage 3, and 21.4% and 34.3% for stage 4, respectively. When analyzed using the current International Union Against Cancer (UICC) pathologic (p) TNM staging system, the 1-year and 3-year DFS rates were 86.2% and 64.0% for stage 1 (n = 30), 73.9% and 50.0% for stage 2 (n = 182), and 46.8% and 22.3% for stage 3 (n = 111), respectively. Our results showed that, while both staging systems allow clear stratification of patients into prognostic groups, the modified TNM system is not superior to the UICCpTNM system in predicting survival of HCC patients after curative partial hepatectomy. A larger scale, multicenter study may be needed to test the revised TNM system. PMID- 10389370 TI - Endoscopic ultrasonography in the differential diagnosis of giant gastric folds. AB - Giant gastric folds (or large gastric folds) are found in both benign and malignant diseases, and differential diagnosis with either upper gastrointestinal X-ray or endoscopy is difficult. Sometimes, even endoscopic biopsy cannot establish a definitive diagnosis. Recently, endoscopic ultrasonography (EUS) has been used to study giant gastric folds. We performed EUS in 25 patients with giant gastric folds that had been detected with upper gastrointestinal X-ray or endoscopy. The definitive diagnoses were confirmed by histopathology, other examinations, or long-term follow-up. The final diagnoses of these 25 patients were gastric varices in eight, gastric lymphangiectasis in one, gastritis in four, gastric carcinoma (scirrhous type) in six, and gastric lymphomas in six. All patients with gastric varices had anechoic tortuous varicose veins in the submucosal layer. EUS images of gastric lymphangiectasis were similar to those of gastric varices. EUS revealed regular gastric wall thickening of the second (mucosa) and third (submucosa) layers in all cases of gastritis. The fourth (muscularis propria) layer was intact in the only case of mucosa-associated lymphoid tissue lymphoma (MALToma), but not in the other five cases of gastric lymphoma. The second and third layers of this MALToma were irregular in thickness and heterogenous in echogenicity, different from the characteristic EUS findings in gastritis. The fourth layer was markedly thickened only in malignant conditions. Differentiation of gastric cancer from lymphoma with EUS was difficult because of overlapping EUS findings. In conclusion, EUS is indicated for the differential diagnosis of giant gastric folds. In addition, it avoids the risk associated with biopsy of gastric varices. PMID- 10389371 TI - Nonsurgical transvenous retrieval of fractured implantable central venous access device. AB - Implantable central venous access devices (ICVADs) are commonly used in patients with cancer and chronic diseases. Fracture and distal embolization of an ICVAD fracture is rare. A total of 1,006 central venous access procedures using ICVADs were done in National Taiwan University Hospital from January 1994 to December 1996. ICVAD fractures were noted in five of these 1,006 cases. All the fractured catheters were retrieved successfully by nonsurgical transvenous methods. The clinical manifestations were nonspecific palpitation or chest pain. The fracture site was always on the crossover point of the clavicle and the first rib on chest radiography. A pinch-off mechanism was suspected, and supported by scanning microscopy imaging. A deformed and elliptical shape of the fractured catheter site resulted from repeated wear and tear by the movement of the clavicle and first rib and associated muscle groups. The clinical course was benign if early retrieval of the distal fractured catheter by snare loop was possible. A more lateral position by a subclavian approach or an axillary approach for central venous access may avoid this procedure-related complication. PMID- 10389372 TI - Efficacy of pelvic floor rehabilitation for treatment of genuine stress incontinence. AB - To assess the clinical efficacy of a pelvic floor rehabilitation (PFR) program for treatment of genuine stress incontinence (GSI), we studied 72 patients with slight to moderate (2-10 g of urine loss per hour) or severe (11-50 g of urine loss per hour) GSI who underwent PFR. Objective and subjective assessments were performed before and 3, 6, 12, 18, and 24 months after the start of treatment. The overall success rate (complete cure or marked improvement in symptoms) was 61% (44/72) at the 2-year follow-up. The number of leakages per 24 hours and urine loss in the 1-hour pad test were significantly reduced, and vaginal muscle strength was significantly increased in successfully-treated patients. Significant changes were also observed in symptoms of micturition frequency and nocturia and in volume at first desire to void during cystometry in the treatment success group. Patient compliance with the exercise program was a significant predictor of success. The success rate during the 2-year follow-up period, estimated according to patient compliance, also differed significantly among groups, with good, moderate, and poor compliance. Patients experienced no serious adverse effects. These results show that the PFR program used in this study is an effective alternative to surgical intervention for the treatment of GSI in selected patients. PMID- 10389373 TI - Dominant-inherited hypokalemic periodic paralysis in a large Chinese family. AB - Familial hypokalemic periodic paralysis (HoPP) is a rare condition among Chinese. We studied a large Chinese family (48 members in six generations) with dominant inherited HoPP, using incidental tracing of family history of a proband who presented with the typical features of HoPP. Fifteen family members were found to have the disease. We found the familial type of HoPP to differ from the sporadic type in Taiwan, in that the familial type has an equal gender distribution, earlier onset of paralytic attacks, and more severe clinical features in both frequency and extent. When these patients were compared with Caucasian families, the common features were the involvement of the respiratory and the bulbar muscles, and the eye muscles in Chinese patients. Cold-induced attacks and permanent muscle weakness were not common in Chinese subjects. Age and history of paralytic attacks were not the major determinants for the development of permanent muscle weakness. Two family members died during attacks because of severe involvement of the respiratory and bulbar muscles. PMID- 10389374 TI - Management of pancreatic pseudocysts by endoscopic cystogastrostomy. AB - Open surgical drainage is currently the treatment of choice for pancreatic pseudocysts, but endoscopic transmural drainage is another minimally invasive surgical alternative. In this report, we describe two patients with symptomatic pancreatic pseudocysts treated with endoscopic cystogastrostomy. The first patient, a 15-year-old boy, had an episode of traumatic pancreatitis after abdominal injury from a car accident, and complained of postprandial vomiting and abdominal distention 4 weeks later. A large pancreatic pseudocyst, about 10 cm x 6 cm, was noted. The second patient, a 44-year-old man, had a 1-year history of recurrent alcoholic pancreatitis prior to this admission. He suffered from abdominal distention for several weeks. Sonography revealed a large pancreatic pseudocyst, about 18 cm x 9 cm in size. Both patients underwent successful endoscopic cyst-drainage without recurrence. These cases illustrate that endoscopic transmural drainage provides a minimally invasive and effective approach to the management of pancreatic pseudocysts. PMID- 10389375 TI - Laser flare-cell meter analysis of blood-aqueous barrier functional status in a patient with crystalline retinopathy. AB - This is the first report in the literature of the application of a laser flare cell meter to examine the functional status of the blood-aqueous barrier in a patient with crystalline retinopathy. The patient, a 34-year-old man with typical crystalline retinopathy, had crystal deposits scattered throughout the posterior pole of the fundus in both eyes. Fluorescein angiography showed atrophy of the retinal pigment epithelium and geographic areas of choriocapillaris atrophy in the posterior pole. Electroretinography showed subnormal amplitude, and the light peak/dark-trough ratio of the electrooculogram was reduced. Laser photometry showed an increase in the aqueous flare intensity in both eyes, compared with normal values obtained from healthy control subjects. The increase in the aqueous flare intensity in this patient indicates that the function of the blood-aqueous barrier might have been affected in crystalline retinopathy. PMID- 10389376 TI - Tricuspid atresia with persistent truncus arteriosus. AB - The association of tricuspid atresia and persistent truncus arteriosus is a very rare congenital anomaly. We report a newborn with a prenatal diagnosis of tricuspid atresia, in whom associated type II persistent truncus arteriosus was found by postnatal echocardiography. The patient had mild cyanosis and developed heart failure soon after birth. Balloon septostomy was performed to enlarge the interatrial communication. However, her condition rapidly deteriorated and she died of sepsis and heart failure at the age of 14 days. PMID- 10389377 TI - Long-term outcome of infants at the margin of viability. AB - The outcome of babies at extremely short gestational age (22 to 26 weeks) effects our clinical decisions regarding their care. We looked at survival and presence of disability at 25 +/- 11 months of age in 246 of these infants born at our hospital between 1992 and 1996 who were average weight for gestational age. Babies were evaluated in our follow up clinic by a pediatrician, and a physical therapist for cerebral palsy, blindness and deafness, and by a psychologist with the Bayley II. Chances for survival without disability exceeded 50% of live born infants at 25 weeks gestation or a birth weight of 700 to 800 grams. Chances for survival exceeded 50% of live born infants at 24 weeks gestation or a birth weight of 600 to 700 grams. Chances for intact survival reached 50% of survivors at 23 weeks gestation or a birth weight of 400 to 500 grams. PMID- 10389378 TI - Selected facts about teen pregnancy in Mississippi. PMID- 10389379 TI - The Year 2000 Problem: guidelines for protecting Your patients and practice. American Medical Association. PMID- 10389380 TI - A message from HCFA. Health Care Financing Administration. PMID- 10389381 TI - Nancy O'Neal Tatum, 1950-1998: a beautiful, blessed life. PMID- 10389383 TI - Medicare update. PMID- 10389382 TI - Information and Quality Healthcare update. PMID- 10389384 TI - The modifiable factors contributing to leading causes of death in South Carolina. AB - In 1996, there were 34,035 deaths in South Carolina. Almost 70 percent of these deaths were due to chronic diseases. There are known ways to prevent chronic diseases from developing or at least delay their developmental process, thereby lengthening years of life. The purpose of this paper is to report modifiable risk factors for mortality related to leading causes of death. The top ten causes of death in South Carolina were obtained from the South Carolina Department of Health and Environmental Control. Estimates of the number of deaths due to certain modifiable risk factors were made using results of a study published by McGinnis and Foege. The percentage of deaths due to each cause was adapted to South Carolina death certificate data from the national estimates. Results indicate that small modifications in individual lifestyles could prevent or delay nearly 50 percent of deaths in South Carolina annually. Tobacco use, diet and physical activity, and misuse of alcohol contribute to the largest number of deaths. Other modifiable behaviors contributing to the 50 percent mortality are microbial agents, toxic agents, firearms, sexual behavior, motor vehicle accidents, and illicit use of drugs. The implication in these findings is that these risk factors for mortality are mainly modifiable. There are many causes of death that may be delayed due to these modifiable risk factors. By looking at preventable causes of death, rather than focusing on traditional causes of death, it becomes clear that prevention strategies are critically important. PMID- 10389385 TI - Fire ants: a continuing community health threat in South Carolina. AB - Imported fire ants are now firmly established in all 46 counties of South Carolina. In 1998 there were an estimated 660,000 cases in the state of which approximately 33,000 sought medical treatment at an estimated cost of 2.4 million dollars. Residents and visitors are at risk for IFA attacks that may occur indoors as well as outdoors. While IFA sting victims in endemic areas of the state may be less likely to seek medical treatment, patients in recently infested areas seem to be more likely to seek treatment since they are relatively unfamiliar with the multiple, painful IFA stings and pustules. Citizens need to control IFA infestations around and within their homes according to guidelines recommended by the Clemson University Cooperative Extension Service. Regional IFA control programs were discontinued in the past because of costs and environmental chemical concerns. Obviously, we need to support ongoing research aimed at developing improved and safe strategies for the local and regional control of IFA. PMID- 10389386 TI - Promoting health in South Carolina: a charge to primary care providers. PMID- 10389387 TI - Hearing impaired patients. Legal obligation to treat and pay for interpreters. PMID- 10389388 TI - One of MSU's best-kept secrets. Free CME boosts knowledge of neurology. PMID- 10389389 TI - [The problem of consent in anesthesia and intensive care. The proposal of SIAARTI]. PMID- 10389390 TI - Levels of evidence for the pharmacological effectiveness of prolonged methylprednisolone treatment in unresolving acute respiratory distress syndrome. PMID- 10389391 TI - Clinical and technical approach to selective decontamination of the digestive tract (SDD). PMID- 10389392 TI - Alkali therapy in the treatment of acute metabolic acidosis. PMID- 10389393 TI - Lactic acidosis. A new insight? AB - From an intensivist point of view, lactic acid is (i) responsible for metabolic acidosis, (ii) related to anoxia or ischemia and (iii) associated with poor prognosis. Conversely, from a biochemist point of view lactate is a good cellular substrate which can be easily converted to pyruvate and used as gluconeogenic substrate, or oxidised or transaminated into alanine. Hence the main question is not anymore to assess the value of lactate concentration as a marker of severity (it is well established) but rather to understand the metabolic meaning of its increase: is it beneficial or deleterious? In fact several recent experimental works have shown that instead of being a negative consequence, lactate production and related metabolic acidosis due to the stimulation of anaerobic ATP-production pathway could be a protective adapted response. PMID- 10389394 TI - Mechanism of action of anaesthetic drugs. PMID- 10389396 TI - Intravenous infusion techniques: how to do it and why we should do it. PMID- 10389395 TI - Drug interactions in anaesthesia. AB - Anaesthesia is nowadays seldom accomplished by a single agent because no single agent is able to provide all components of anaesthesia without seriously compromising haemodynamic and/or respiratory function, reducing operating conditions, or postponing postoperative recovery. Because of the small therapeutic window a detailed characterisation of the concentration-effect relationships of anaesthetic agents is required to allow a proper selection of the various intravenous agents and the combinations thereof to an optimal therapeutic pharmacological effect in the absence of significant side effects. During the past decade, for propofol and the various opioids fentanyl, alfentanil, sufentanil, and remifentanil considerable progress has been made in the characterisation of the pharmacokinetics and pharmacodynamics of these agents and of the combinations thereof. This manuscript describes the pharmacokinetic and dynamic interactions between these agents and the determination of optimal concentrations that assure adequate anaesthesia and a rapid return of recovery. PMID- 10389397 TI - Circulatory effects of volatile anesthetic agents. AB - Modern general anesthesia includes the use of multimodal techniques, mixing volatile or intravenous agents with opioids and/or regional anesthesia. Therefore, most circulatory side effects of inhaled agents are seldom observed. They all produce dose-dependent decrease of systolic and diastolic function and depress baroreflexes to a varying degree. Desflurane, and to a much lesser degree, isoflurane, may cause sympathetic nervous system activation when inhaled concentration is rapidly increased to above 1.5 MAC. This appears to be the result of central activation rather than airway irritation. The older volatile anesthetics halothane and enflurane have no or minimal effect on coronary vascular tone. In contrast, the more recent agents all limit coronary vasodilator reserve by direct effect upon coronary resistance vessels, sevoflurane less than isoflurane and desflurane at equipotent anesthetic dose. Although isoflurane and desflurane appear to produce myocardial ischemia by non-hemodynamically related mechanisms more frequently than halothane or fentanyl in patients at risk, no particular agent has been identified to carry greater risk for adverse cardiac outcome than others. PMID- 10389398 TI - Why, how and when to monitor neuromuscular function. AB - BACKGROUND: Traditionally, anaesthetists evaluate the effect of neuromuscular blocking agents clinically. We observe the fasciculations following injection of succinylcholine, the movements of the reservoir bag, the spontaneous movements of the patient, headlift etc. However, with the advent of new fast acting neuromuscular blocking agents and the increasing awareness of the problems of postoperative residual neuromuscular block there is an mounting understanding of the importance of a more objective assessment of the neuromuscular function during anaesthesia. PURPOSE OF LECTURE: In this lecture I shall give my personal bias on whether or not routine monitoring of neuromuscular function during anaesthesia is essential. Also, I shall try to answer the question "why, how and when should we monitor neuromuscular function during clinical anaesthesia?" PMID- 10389399 TI - Epidemiology, mechanisms and clinical features of rhabdomyolysis. PMID- 10389400 TI - Acute renal failure in rhabdomyolysis. AB - Fifteen to 30% of patients develop acute renal failure (ARF) following rhabdomyolysis and rhabdomyolysis accounts for 5 to 9% of all ARF. Experimental studies revealed two critical factors that predispose to myoglobinuric ARF: hypovolemia/dehydration and aciduria. At the nephron level, three basic mechanisms underlie heme protein toxicity: renal vasoconstriction with decreased renal blood flow, intraluminal cast formation and direct heme protein-induced cytotoxicity. During the early phase of myoglobinuric ARF, hemodynamic process are mainly involved in glomerumar filtration rate decrease while tubular mechanisms occur in the late phase. Critical factors which predispose to myoglobinuric ARF in animal models--i.e. hypovolemia/dehydration and aciduria- are also encountered in human epidemiological studies. Prevention of myoglobinuric ARF rely on rapid and adequate correction of fluid deficits with saline, bicarbonates and mannitol. The choice of hemodialysis technique in the case of constituted ARF strongly depends on the site of intervention, especially in the case of rescue operation. The care of myoglobinuric ARF in intensive care unit do not differ from this of ARF from other causes. PMID- 10389401 TI - The management of shock and local injury in traumatic rhabdomyolysis. AB - Rhabdomyolysis (literally "striped muscle dissolution") is a biological and clinical condition that takes to plasmatic release of myoglobin, muscle enzymes and electrolytes, relates to the lysis of stripped muscle fibers. Rhabdomyolysis presents the clinician with two distinct problems: local injury and the systemic effects directly related to that injury. Locally, muscle, vessel and nerve compression are the primary issues. Systemic concerns relate to depleted intravascular volume, electrolyte imbalances and renal injury from myoglobin. Preventing the systemic and renal complications of the crush syndrome requires very early and vigorous treatment to sustain the circulation, preferably started at the site of the catastrophe. During the extrication of an injured person from a collapsed building, wrecked automobile, or other site, isotonic saline solution should be infused at the rate of 1.5 liters per hour as soon one of the trapped person's limbs has been freed. Some authors suggest to do a preventive fasciotomy in any suspicious case of compartmental syndrome, when the patient has severe muscular pain of the muscular cavity, tense swelling, hypoesthesia or anesthesia of the muscular cavity, pain at the passive mobilization of the limb. On the other hand other surgeons suggest doing a fasciotomy only in selected group of patients. Therefore, the traumatic rhabdomyolysis has few diagnostically problems. On the other hand, their treatment is complex and must have a multidisciplinary approach. So the rhabdomyolysis actually remain a severe disease with high mortality caused principally by visceral lesions related to sepsis. PMID- 10389402 TI - [Mechanical ventilation during thoracic anesthesia]. AB - Aim of the study was to test individual mechanical and functional responses to open chest lateral decubitus during one lung ventilation. We measured dependent lung pressure volume (P-V) curves of 19 patients during supine and lateral decubitus. We found that patients characterized by high FEV1 developed greater changes in P-V curve shape than those characterized by low FEV1. Based on these results we decided to test a ventilation strategy characterized by the use of ZEEP or PEEP = 10 cm H2O applied to the dependent lung. In a preliminary set of patients stratified by FEV1 we found that PEEP deteriorated PaO2/FiO2 in patients with low FEV1, while there was a trend towards improvement in patients with high FEV1. It is possible that dependent lung PEEP counteracts atelectasias in normal lungs, while it may divert blood flow or create dead space in patients with sick and stiff lungs. We conclude that during one lung ventilation in open chest lateral decubitus, ventilatory setting need to be individually tailored. PMID- 10389404 TI - One lung ventilation: prospective from an interested observer. AB - The improvements in video endoscopic surgical equipment and a growing enthusiasm for minimally invasive surgical approaches, brought video assisted thoracoscopy (VAT) to the practice of surgery for diagnostic and therapeutic procedures. Most of these procedure required a well collapse lung and should be included in the absolute indication for one lung ventilation (OLV) category. The univent tube, is a novel means of achieving bronchial blockade. The bronchial blocker technique has been modified so that the bronchial blocker is passed along a single-lumen endobronchial tube. It was introduced to clinical practice to avoid the need to change the double lumen tube at the conclusion of the procedure. Finally, one of the most interesting future concept to keep adequate oxygenation during OLV, is the ability to modulate the lung circulation. In fact inhaled nitric oxide (NO) and intravenous Almitrine have been combined with additive effects on gas exchange. In case of OLV using that combination will maximize the HPV of the non dependent lung while dilate the dependent lung to practically eliminate the transpulmonary shunt. PMID- 10389403 TI - Post-thoracotomy analgesia and perioperative outcome. AB - Post-thoracotomy pain is the most severe form of pain after surgery and is continuously exacerbated by ventilatory function. Due to the multiplicity of nociceptive inputs from the chest wall, thoracic viscera, diaphragm and postoperative chest tubes, postoperative pain may be difficult to control with single modalities. The aim is excellent analgesia with function i.e. normal ventilation and rapid mobilisation. A variety of agents and techniques have been shown to be effective analgesics with varying degrees of functional success. These include systemic opioids, NSAIDS and ketamine, regional analgesia (including epidural, spinal, paravetebral, intercostal and intrapleural techniques) and cryoanalgesia. The most popular and probably most effective technique at the present time is thoracic epidural analgesia using a combination of different local anesthetic agents and opioids. There are few data indicating any influence on outcome of different postthoracotomy analgesic techniques. Improvement in outcome requires a co-ordinated approach from all caregivers using the best possible analgesic techniques. PMID- 10389405 TI - Fluid replacement in ICU. PMID- 10389406 TI - Transfusion risks and limitations. AB - The safety of the blood transfusion therapy has dramatically increased over the last few years because of improvements in donor screening, testing of donated blood and pre-transfusion tests. However blood transfusion can never be seen as a risk free procedure. The risks to which the patients receiving blood are exposed are infectious, immunologic and other non infectious, non immunologic hazards. Transmission of viral, bacterial and protozoal infections is probably the greatest concern associated with allogeneic blood transfusion. While the risk of transmitting viruses is now very small, there is an increasing concern regarding bacterial contamination of donated blood. Among immunological sequelae, beside alloimmunization, are fever and chills, allergic and acute hemolytic reactions, the last being the currently most important cause of deaths associated with blood transfusion. Moreover allogeneic blood transfusion may lead to immunosuppression, which may increase the risk of infection and cancer recurrence. Other non infective pulmonary edema and physical and biochemical alterations (such as hypothermia, electrolyte and acid base disturbances). PMID- 10389407 TI - Anemia and red cell transfusion in critical care. Transfusion Requirements in Critical Care Investigators and the Canadian Critical Care Trials Group. PMID- 10389408 TI - Hydroxyethyl starch: pharmaceutical and clinical profile. PMID- 10389409 TI - Clinical perspectives of "the open lung concept". PMID- 10389410 TI - [The sigh in ARDS (acute respiratory distress syndrome)]. AB - We studied 10 consecutive, sedated and paralyzed patients with Acute Respiratory Distress Syndrome (ARDS). The entire study lasted 4 hours, divided in 3 periods: 2 hours of recommended ventilation [lung protective strategy, LPS, i.e., ventilation with low tidal volume (< 8 mL/kg), limiting the plateau at 35 cm H2O, together with high positive end-expiratory pressure (PEEP)], 1 hour of sigh (LPS with 3 consecutive sighs/min at 45 cm H2O plateau pressure), and 1 hour of LPS. Total minute ventilation, PEEP, FiO2 and mean airway pressure were kept constant. The introduction of sighs induced a consistent recruitment and PaO2 improvement, and a decrease in venous admixture and PaCO2. Interrupting sighs and resuming LPS led to a progressive derecruitment, and all the physiological variables returned to baseline. Derecruitment was higher in patients with higher PaCO2 and lower VA/Q ratio. We conclude that: 1) LPS alone does not provide full lung recruitment and best oxygenation in ARDS; 2) application of sigh may provide pressure enough to recruit and volume enough to prevent reabsorption atelectasis. PMID- 10389411 TI - [What kind of monitoring of intracranial pressure]. AB - Intracranial pressure monitoring is an essential element of severe head injury monitoring. In the Italian reality, this monitoring still has not become diffusely utilised. In this paper are analysed, using the available data obtained from the Neurolink database, the information from the more advanced Italian neurotrauma centres. Only 50% of the patients, in which monitoring is indicated, are monitored. Synthetically will be reviewed the indications to the monitoring, the advantages offers from such procedure, the modalities of monitoring considering also the economic impact and the risks legacies to the procedure. PMID- 10389412 TI - [What will jugular bulb oxygen saturation monitoring tell?]. AB - Global cerebral oxygenation can be measured by means of a catheter introduced in the internal jugular vein and placed retrograde in the jugular bulb. The measure of oxygen saturation sampled from the jugular vein (SjvO2) depends on cerebral metabolism and blood flow. This parameter describes the relative balance between oxygen delivery to the brain and oxygen consumption by the brain. SjvO2 remains normal until cerebral blood flow is proportional to cerebral metabolic demands. Any disturbances that increase cerebral metabolism and/or diminishes cerebral oxygen supply determines a reduction of SjvO2. Correspondingly, a decrease of oxygen consumption and/or an increase of oxygen supply may induce an increase of SjvO2. Therefore, SjvO2 is a useful monitor to assess the adequacy of cerebral circulation in patients with neurologic illness, allowing detection of state of hypoperfusion. Monitoring cerebral oximetry in comatose patients is of great importance in order to prevent, detect, control and understand secondary brain insults and damage which are mainly ischemic/hypoxic in nature. Although SjvO2 was shown to be highly sensitive in the presence of global hypoxia or ischemia, the occurrence of focal ischemia may still go undetected. Besides this, elevated SjvO2 should not be universally interpreted as hyperaemia. Instead, the presence of an elevated SjvO2 is a heterogeneous condition. Increased SjvO2 may be alarming prognostic indicators that carry important implications for comatose patients management. PMID- 10389413 TI - [Cerebral tissue oxygen monitoring: a useful thing?]. AB - Monitoring cerebral oxygenation has been one of the main fields of interest in neurointensive care during the past few years. In fact it is strongly believed that restoring adequate cerebral oxygenation is the premise to maintaining the viability and restoring the function of the damaged CNS. Global monitoring provides an indirect estimation of adequacy of substrates supply to the brain. Local measurement of brain oxygen tension (ptiO2) is possible through a Clark electrode implanted into the cerebral parenchyma. The paper describes the physical basis of the monitoring, the pathophysiology of ptiO2 and its clinical use. PMID- 10389414 TI - [Monitoring in neurotraumatology in -----------]. AB - Monitoring intracranial pressure (ICP) and cerebral perfusion pressure (CPP) is fundamental in avoiding ischemic damage in severe head injury patients. Monitoring CPP can provide prompt informations: decide to start or to stop a therapy, control the effect, give indications for surgical treatment; unfortunately this monitoring is not able to understand the pathophysiology of the raised ICP. In the last years new techniques of monitoring were proposed with the aim to identify the problem and to target therapy: electrophysiological, biochemical, brain oxygenation, ultrasonography monitoring can be usefull in a concept of multimodality monitoring. PMID- 10389415 TI - [What kind of monitoring should be selected when resources are limited?]. PMID- 10389416 TI - [The GiViTI protocol on enteral nutrition: experimental design and parameters of effectiveness]. PMID- 10389417 TI - CPR in the trauma patient: indications and contraindications. PMID- 10389418 TI - Chest radiography: potential and limitations. PMID- 10389419 TI - Proportional assist ventilation. PMID- 10389420 TI - [The politics of antibiotics in the hospital: the role of the pharmacist]. PMID- 10389421 TI - [Pre- and postoperative treatment of hypertensive crisis]. PMID- 10389422 TI - [Physiopathology and clinical aspects of hypertensive crisis]. PMID- 10389423 TI - [Privacy in intensive care]. PMID- 10389424 TI - [Consequences of perioperative hypothermia]. PMID- 10389425 TI - [Level 1 systems in clinical practice]. PMID- 10389426 TI - [One day anesthesia in pediatrics]. AB - The paper includes a short review on paediatric anaesthesia in day surgery. It discussed on why day surgery in children is so popular since the beginning of the century; the Italian laws on this item and the guidelines of the Italian Society of Anaesthesiology on "day surgery" and "preoperatives of children". PMID- 10389427 TI - [New curare agents in pediatric anesthesia]. PMID- 10389428 TI - [Remifentanil in pediatric surgery]. PMID- 10389429 TI - [COPA, LMA, tracheal intubation. What are the indications in pediatric anesthesia]. PMID- 10389431 TI - [Invasive ventilation in pediatric patients discharged from intensive care to continuous home care]. PMID- 10389430 TI - [Control of postoperative pain in pediatric patients]. PMID- 10389432 TI - [Anesthetic problems in the mediastinal masses]. PMID- 10389433 TI - [Problems in anesthesia in pulmonary cystic adenomatosis]. AB - Congenital cystic adenomatoid malformation (CCAM) of the lung is a rare disease. It is about an abnormal proliferation of mesenchymal elements and failure of maturation of bronchiolar structures, characterized by the replacement of normal pulmonary tissue with "cysts" in variable size and number. These lesions communicate with the tracheobronchial tree. During fetal period hydrops and polyhydramnios can be associated with CCAM. A cystic adenomatoid malformation can be detected by antenatal ultrasound, and, at the birth, it is confirmed by chest radiography. From January 1990 to December 1998, 24 cases with CCAM came to our observation; 16 of these patients underwent surgery and 14 have come to a complete recovery. The newborns, with CCAM, can show early acute respiratory distress for rapid expansion of the cysts leading to compression of normal lung and mediastinal shift. Conventional mechanical ventilation may cause further expansion of the involved lobe with a ball-valve effect: this take a clinical deterioration. Perioperative ventilatory management with high frequency oscillation (HFO) is useful to stabilize and to improve arterial blood gases of this patients. At the moment, thanks to the early prenatal sonographic diagnosis, it is possible, and strongly advisable, after adequate serial checkings during the pregnancy, to refer CCAM cases to a tertiary centre that is properly equipped, where a poly-specialist team consisting of obstetrician, neonatologist, pediatric anesthetist and pediatric surgeon, will be able to plan and arrange in the best treatment necessary for the newborn. PMID- 10389434 TI - [Inhalation of foreign bodies]. AB - Accidental aspiration of a foreign body (FB) is an event which is reasonably frequent and dramatic in children and is still today one of the main causes of death due to accidents at home in children up to three-four years of age. The severity of the clinical picture varies according to the size, shape, type and site of arrest of the material aspirated and can be associated with both severe asphyxial forms and forms with insidious and vague symptoms which are difficult to diagnose correctly. A late diagnosis is however a fairly common event in literature. An anamnesis suggesting probable aspiration in a child under the age of 3 should direct doctors towards diagnostic and operative endoscope examinations of the patient, even where there is a negative clinical and radiological picture. Organic material, mainly peanuts, represented 60-75% of the findings, particularly in the 0-3 year age-band. In the other of cases inorganic material was extracted from school-age children. Aspiration of a FB exposes the patient to risk of serious complications and sequelae. Antibiotic, dexamethasone therapy and the ventilation support in the CPAP helped to avoid post-extractive sequelae. Prevention should in any case be the primary aim as regards to aspiration of foreign bodies in children. This should be stimulated by appropriate educational campaigns to raise awareness. The study included 62 child patients observed in the Department of Anesthesia and Intensive Care of the S.Orsola-Malpighi Hospital of Bologna over the last 11 years who were admitted for suspected FB aspiration. PMID- 10389435 TI - Natural surfactant supplementation in ARDS in paediatric age. AB - OBJECTIVE: To evaluate the effects of natural surfactant supplementation in infants, children and adolescents affected by ARDS from different origins in order to reduce lung barotrauma due to artificial ventilation, improve gas exchange, reduce oxygen toxicity and survival. MATERIALS AND METHODS: Two groups, the first consisting of 22 children, 7 days-24 months, and the second of 8 oncohaematologic patients, 2-16 years, affected by ARDS from sepsis, inhalation syndrome and interstitial pneumonia, candidates for ECMO, were treated intratracheally with 50 mg/kg of natural surfactant. Before treatment all patients had been mechanically ventilated using PEEP levels > or = 8 cm H2O and FiO2 > or = 0.6, for at least 24 hours without any improvement in gas exchange. RESULTS: From 15 mins after surfactant administration a progressive improvement in PaO2 was noted which peaked at 3 hours. In two cases in the first group a worsening in PaO2 occurred starting from 12-18 hours, which needed additional doses. All patients in the second group needed additional doses after 12 h. No significant PaCO2 variations were noted until 24 hours. In all cases the chest X ray improved at 4 hours and clearing was obtained starting from 24 hours in those cases where an additional dose had not been necessary. Computed Tomography confirmed the improvement in lung pathology. All the children in the first group survived except one HIV-positive child. The oncohaematologic children showed an improvement in PaO2 after each administration of surfactant even though they later died due to their initial disease, except one child. COMMENT: Surfactant efficacy in this study appears to depend on the severity of lung pathology and to be strictly connected with early treatment. PMID- 10389436 TI - [Enteral nutrition in critical patients]. PMID- 10389437 TI - [After the abolition of the code of professional conduct, what are the prospects for autonomy?]. PMID- 10389438 TI - [Anxiety and stress in the nursing staff. A comparative study between intensive care and general wards]. AB - We studied a population of 463 nurses working in intensive care units--ICUs- (distributed in 51 italian hospitals), and 216 nurses working in general medicine units (distributed in 17 italian hospitals). They we asked to fill in a form including: 1) general data and his/her work environment, and 2) some standardized scales (HAD A and D, STAI Y-1 and Y-2, MBI) for estimation of anxiety, depression and "burnout" syndrome. We used also the "P questions", evaluating the different situations of work environment causing anxiety. The aims of the study were to evaluate the effectiveness of different scales and the influence of general medicine and intensive care environment on psychological features. Among different scales estimating anxiety, the STAI Y-2, valuing a chronic anxiety status, results to be efficient, beside the already tested HAD A. The intensive care environment did not seem to be more stressful for nurses staff rather than general medicine units. Furthermore, nurses operating in general medicine units have a major tendency to depression; their work environment seems to favour the development of anxiety. Finally, it results that general medicine units cause a more severe "burnout" syndrome in their nurses staff rather than ICUs. PMID- 10389439 TI - [A new technique for pronation. Benefits for patients and operators]. PMID- 10389440 TI - [Personal comments on decubitus prophylaxis by the nursing staff in the resuscitation and intensive care unit at A. Cardarelli Hospital in Naples]. PMID- 10389441 TI - [EurIuS II (European Intensive Care Units Study). The effect of the harmonization and standarization of nursing in intensive care in the European Community]. PMID- 10389442 TI - [Handling and assistance. How to prevent vertebral pathologies]. PMID- 10389443 TI - Surgical and endovascular treatment of stenosis of the innominate artery. AB - BACKGROUND: Atherosclerotic lesions of the brachiocepfialic trunk are relatively rare compared with other types of vascular diseases. Median sternotomy with direct endothoracic repair is recommended because of good early and long-term results. Nevertheless, this procedure is not without risks such as hemorrhaging, embolism, aortic dissection, infection or death. METHODS: This report therefore, describes our experience in intraoperative balloon angioplasty and additional stent placement of isolated stenosis of the brachiocephalic trunk with cerebral protection ensured by common carotid artery clamping. Through an anterolateral cervical approach the right common carotid artery was surgically exposed. After dilating the brachiocephalic trunk and positioning the stent, the vessel was repaired with a continuous suture. RESULTS: In all patients the dilation of the stenosis of the brachiocephalic trunk and the stent placement were successful without any side-effects. No distal embolism with neurologic events, innominate artery dissection, rupture, occlusion or neck hematoma occurred. All patients were discharged three days after the intervention. CONCLUSIONS: This technique offers a safe, effective approach to stenoses of innominate arteries because it is less invasive than conventional transthoracic or extrathoracic surgery and offers excellent early and mid-term results. We believe that this technique is a safe and effective alternative to conventional surgery. PMID- 10389444 TI - Study of thrombinic activation indexes, in patients with lower limb critic ischaemia, before and after prostaglandin E1 therapy. AB - BACKGROUND: In this study the action of a prostaglandin, PGE1, was studied in a group of patients with peripheral arterial occlusive disease (PAOD). METHODS: In 16 patients (14 men and 2 women, aged 47-70 years, mean 57 +/- 7) with PAOD, Fontaine stage IIb and III in critical ischemia, the effects on two indexes of thrombin generation and action of the endovenous administration (2 hours) of 60 micrograms of Alprostadil-PGE1 for four weeks were evaluated. In all artheriopathic patients, before and after pharmacological treatment, the following haemostatic parameters were evaluated: the prothrombin fragment 1 + 2 (F1 + 2) and the fibrinopeptide A(FPA). RESULTS: The patients showed plasma levels of FPA significantly decreased at the end of the treatment. On the other hand, no significant difference in plasma F1 + 2 levels was observed after treatment. CONCLUSIONS: These results seem to indicate that plasma F1 + 2 levels are significantly elevated, as a marker of thrombosis status, in patients with PAOD before and after treatment with PGE1. PMID- 10389445 TI - [Planar myocardial scintigraphy with 99mTc sestamibi for the evaluation of the infarct area and the efficacy of the thrombolytic therapy]. AB - BACKGROUND: Nuclear cardiology permits the estimation of myocardial infarction size and the result of the thrombolytic therapy. The aim of the study was to demonstrate the feasibility of the planar myocardial scintigraphy with tecnetium 99m-sestamibi in the coronary intensive care unit for the early identification of the infarct size and the results of the thrombolytic therapy. MATERIAL AND METHODS: We studied 15 patients affected by a first acute myocardial infarction (AMI), 10 anterior and 5 inferior wall, treated with thrombolysis (APSAC 30U i.v.) within and interval of 3 hours from the symptoms onset, tecnetium-99m sestamibi was injected before thrombolysis and after 3 +/- 1 hours the planar imaging was registered with a mobile gamma-camera. Scintigraphic evaluation was repeated after 24 hours and before patient discharge. Within 48 hours from the thrombolytic therapy the coronary angiography was performed for the demonstration of patency of the infarct-related artery. The left ventricle myocardial perfusion was divided in the 3 planar projections into 13 segments. The perfusion in each segment was evaluated with a perfusion score: 0 = normal, 1 = moderately reduced, 2 = severely reduced, 3 = absent. The sum of the hypoperfused segments represented the infarct size. A perfusion score improvement greater than 40% was considered a marker of reperfusion. RESULTS: The infarct size involved 4.2 +/- 1.5 segments in the anterior and 2 +/- 0.8 segments in the inferior wall infarctions (p < 0.05). The scintigraphic imaging made 24 hours after AMI allowed the diagnosis of coronary reperfusion in 10 patients. The coronarography demonstrated the infarct related artery patency in 14 patients. The nuclear imaging at patient discharge provided the diagnosis or reperfusion in 11 cases and demonstrated an improvement of the myocardial perfusion score in 8 cases. CONCLUSIONS: In patients with AMI treated with thrombolysis the scintigraphic imaging with tecnetium-99m-sestamibi is feasible with a mobile gamma-camera in the intensive coronary care unit. The quality of planar imaging is good and allows the evaluation of myocardial infarct size and the efficiency of thrombolytic therapy. An earlier scintigraphic imaging should be taken into consideration for a more timely non-invasive evaluation of patients who need coronary angiography and, if necessary, a rescue-PTCA. PMID- 10389446 TI - [Segmental vasculitis. Report of a case involving the popliteal artery]. AB - A case of occlusion of the popliteal artery, in a young female patient with limb threatening ischemia, probably due to a segmental vasculitis, is reported. The surgical treatment, consisting of a femoro-distal bypass, was able to improve dramatically tissue perfusion; however, due to septic complications, major limb amputation was unavoidable. It is stressed that the treatment of vasculitis should be adapted to each individual case; in selected patients, surgical revascularization appears to offer the only possibility of correcting a "desperate" situation. PMID- 10389447 TI - [An unusual episode of angina]. AB - The authors report the case of a 77-year-old woman suffering from ischemic cardiopathy, arterial hypertension and NID diabetes mellitus who was hospitalised for diabetic retinopathy. Following the topical administration of a drop of phenylephrine in the right eye, in preparation for fluoroangiography, she suffered an attack of angina pectoris. This event was clearly identified by a ECG carried out at the time which highlighted the sublevelling of the ST tract in the precordial derivations V2-V5, and by ecocardiographic imaging that showed septo apical, anterior-apical and anterior-median transient segmentary hypokinesia. This case underlines the need for prudence and emphasises the need to regard patients suffering from ischemic cardiopathy as high-risk when undergoing diagnostic tests, in particular if these involve the use of vasoactive drugs. PMID- 10389448 TI - Primary lymphomas of the heart in immunocompetent patients. Presentation of two cases with one 12 year survival. Analysis of surgical implications. AB - Two primary malignant lymphomas originating in the right heart cavities have been diagnosed and treated in our department: one, with additional right atrial and inferior vena cava thrombosis required emergency thrombectomy, incomplete excision and chemotherapy and survived 12 years, the other with extensive right ventricular infiltration and failure, died from mediastinitis and aplastic anaemia following surgery and chemotherapy. Quick diagnosis and treatment are indicated. If possible, surgery should be avoided. PMID- 10389450 TI - Banking for tomorrow. PMID- 10389449 TI - [Critical ischemia in elderly patients. Evaluation of the effect of two different methods of Iloprost therapy on the efficacy, tolerance, modification of quality of life and self-sufficiency]. AB - BACKGROUND: The use of Iloprost in the treatment of critical leg ischemia in very old patients can lead to tolerability problems, related to the drug used and to the kind of patient treated. The aim of this study was to evaluate the impact of this therapy on the activities of daily living and on the quality of life of the patient together with its efficacy and tolerability. METHODS: We studied 20 subjects (mean age 74 +/- 6.8) divided in 2 groups homogeneous for age, seriousness of the disease and presence of diabetes mellitus. In the first group Iloprost was administered for 6 hours, for 28 consecutive days, in the second group for 6 hours, two times a day for 14 consecutive days. In each group we observed the following parameters before and after treatment: clinical evaluation of pain and use of analgesics, ADL and SK39 questionnaire, ankle/arm index c.w. Doppler, strain gauge plethismography of lower limbs, TcPO2 at the back-feet level. RESULTS: Treatment was well tolerated in both groups where we observed a similar reduction of pain, a reduction in the instrumental indexes which express the microcirculatory activity, an improvement in the quality of life and in the capacity to carry out everyday activities. The double daily administration of Iloprost did not cause any significant side effect in the subjects studied, and a better responsiveness to the pain symptomatology was observed. CONCLUSIONS: Iloprost can be used in the treatment of critical leg ischemia even for very old patients with good tolerability and effectiveness. In the double daily administration no relevant side effect was observed. This approach is to be preferred also in terms of cost-effectiveness. PMID- 10389452 TI - Banking on blood. PMID- 10389451 TI - Dr. Who? PMID- 10389453 TI - Second act. PMID- 10389454 TI - The National Marrow Donor Program. PMID- 10389455 TI - The new Lyme disease [correction of virus] vaccine. What Minnesota physicians need to know. PMID- 10389456 TI - Twelve years of emergency medicine at Hennepin County Medical Center. Changing critical care experience. AB - OBJECTIVE: To elucidate the critical care experience of emergency medicine faculty and delineate changing patterns of practice. METHOD: A retrospective review of the resuscitation room dictations for all patients treated in the emergency department resuscitation room by one emergency medicine physician from July 1, 1985, to June 30, 1997, was performed. RESULTS: A total of 1,325 cases were reviewed. The number of cases of arrhythmia (p < 0.01) and medical cardiac arrest (p < 0.01) significantly decreased over time, while the number of cases involving firearm injury (p < 0.01) increased. The percentage of trauma cases steadily rose from 38.5% in year 1 to 50.2% (p < 0.01) in year 12. Significant decreases in the rate of arterial line placement (p < 0.01), central line placement (p < 0.01), nasotracheal intubation (p < 0.01), and thoracotomy (p < 0.05) have occurred. Significant increases were seen in cardiac ultrasound examination (p < 0.01), rapid sequence induction for intubation (p < 0.01), and the use of paralytic medications (p < 0.01). CONCLUSION: Significant changes have occurred in the type of case and rate of utilization of various diagnostic and therapeutic procedures. PMID- 10389457 TI - Implementation of a subregional trauma system. PMID- 10389458 TI - Film rouge. PMID- 10389459 TI - [Women's medical problems]. PMID- 10389460 TI - [Misunderstanding of medical resource use by women]. AB - By analysing three different Belgian databases, we could demonstrate that whereas women subjectively declare a higher number of diseases, a worse functional state and a higher trend to use medical resources, they are, from an objective point of view, less frequently hospitalized and cause a lower cost, at certain ages, for the Social Security. PMID- 10389461 TI - [Feminization of the medical profession]. AB - Although we can observe a real change in the participation of women in the medical profession, we must ask ourselves what feminisation means. Does it refer to the number of women, or to an eventual change in the content of the profession due to the fact that women are more oriented toward caring, empathy, relationships. The figures for Belgium show a real improvement in the presence of the women but not in the more prestigious specialisations like surgery or internal medicine. PMID- 10389462 TI - [Smoking among women]. AB - The aims of this article are 1) to give a short overview about the main aspects of a growing disease: smoking among women; and more importantly 2) to stimulate the primary care doctors as well as the specialists to participate into an effort of the whole society to fight against this avoidable epidemy which is responsible for millions of deaths and suffering all over the world. As smoking is considered as a personal habit, the doctors do not like to interfere with this choice of "life style". PMID- 10389463 TI - [Women and alcoholism]. AB - As confirmed by many epidemiological studies, alcoholism remains predominant in men. However, female alcoholism has seemed to increase over the last decades, as woman social status as well as cultural stereotypes about women were being modified. Often associated with psychiatric disorders, alcoholism in women has a specific clinical, psychological and social profile. Biological specificities are also of high importance when differentiating female from male alcoholism. PMID- 10389464 TI - [Is it necessary to institute primary preventive treatment in women with hypercholesterolemia?]. AB - Limited data are available in women from randomized trials of lipid-lowering therapy for primary prevention of coronary heart disease (CHD). In women with isolate hypercholesterolemia but no clinical symptoms of CHD and no other risk factors, there is no evidence that lipid-lowering therapy is of benefit. In postmenopausal women with hypercholesterolemia and other risk factors (particularly diabetes and family history of CHD), lipid-lowering therapy may be of benefit. In those women, hormone replacement therapy may be proposed in the treatment of postmenopausal hypercholesterolemia. PMID- 10389465 TI - [Women and cardiovascular diseases, particularly coronaropathies]. AB - Cardiovascular diseases represent the major cause of death in women. If women are early in life relatively protected by their hormones, after the menopause heart disease and stroke become their greatest health threat. Altogether one in three women will die of cardiovascular disease whereas only one in twenty-two will die of breast cancer. The risk factors for cardiovascular diseases are similar in both sexes. It should be noted however that the impact of diabetes on the development of coronary artery disease is markedly higher in woman than in man. In woman the first presenting symptom of coronary artery disease frequently is angina pectoris. In man it is more often an acute myocardial infarction whether it be symptomatic or silent. The various non invasive techniques used for the diagnosis of coronary artery disease may yield results that pose very difficult problems of differential diagnosis in women. Several studies indicate that in the past, probably because of the ignorance by the public and, perhaps, the medical profession of the high prevalence of cardiovascular and coronary artery diseases in women, the latter have not always received an optimal treatment when they suffered an acute ischemic attack. But, in this decade, they have been dramatic changes in the patents of clinical practice related to coronary heart disease in women. Statistical investigations however indicate that thrombolytic therapy appears less efficacious in women than in men whereas beta-blockers administered early in acute MI are of remarkable efficacy in women. It seems generally agreed that the results of revascularization by coronary artery bypass surgery are poorer in women than in men both in terms of operative mortality and morbidity. Initially, the same pessimism was reported for PTCA. However, recent results allow a much more optimistic view. PMID- 10389466 TI - [Cardiac arrhythmias in women]. AB - There exist gender differences in the frequency of the various forms of cardiac arrhythmias. Atrial fibrillation is more common in man in whom it is often a complication of coronary heart disease. Numerous elderly women however present with this rhythm disorder which is often a complication of congestive heart failure or valvular heart disease. Atrial fibrillation carries a high risk of cerebro-vascular embolic accident in female. Symptomatic supraventricular tachycardia and the long QT syndrome, inherited or acquired, are more frequent among women. The latter are also particularly susceptible to develop torsade de pointe as a consequence of therapy with drugs interfering with the potassium ion channels. Sudden cardiac death is more frequent in men than in women even when the latter suffer from coronary heart disease. Sudden cardiac death without structural heart disease is however more frequently seen in women. Nulliparity, tobacco use and alcoholism might be specific risk factors for sudden cardiac death in female. Cardiac arrhythmias may be seen during pregnancy. New onset or aggravated supraventricular tachycardia may be encountered during that period. Ventricular tachycardia in the absence of structural heart disease may also be seen during pregnancy. The treatment of cardiac arrhythmias in pregnant woman remains a challenge for the practising cardiologist. PMID- 10389467 TI - [Anorexia nervosa, a serious disease of young women]. AB - Anorexia nervosa is characterized by a triad comprising anorexia, weight loss, and amenorrhoea. It is typically present in adolescent or young adult women and its prevalence appears to increase in our society. Eating disorders may lead to cachexia and various severe complications. This disease, which appears to be associated to a particular psychological background, is triggered by conflictual relationships, usually within the family. The anorectic patient should be carefully evaluated, as soon as possible, from a somatic, dietetic and psychological point of view. A specific therapeutic approach should be rapidly proposed in a specialized centre, if possible by a multidisciplinary team. The treatment is most often difficult and of long duration. Prognosis remains uncertain in most cases, especially when appropriate care is delayed. PMID- 10389468 TI - [Obesity and women: esthetic preoccupation or medical problem?]. AB - Women more and more frequently refer to medical doctors for treating weight excess. Even if the primum movens is usually a pure esthetic concern, medical aspects should not be neglected. Indeed, if obesity with gynoid adipose tissue distribution is less deleterious than android obesity from a metabolic point of view, severe obesity is frequently associated with cardiovascular risk factors which may hinder the prognosis of these female patients. Other complications are common in obese women, such as oestrogen-related cancers, osteoarticular problems and psychological disturbances. Various therapeutic approaches are available which permit an encouraging weight loss and a rapid improvement of risk factors. Unfortunately, long-term results are often disappointing, essentially because of the difficulty to follow on the long term a strict diet regimen and practice physical exercise and because of the usual unrealistic expectations of the obese women who consult medical doctors. PMID- 10389469 TI - [Premenstrual tension syndrome or premenstrual dysphoria]. AB - Premenstrual Tension Syndrome (PMS) has always existed: it has been first described by an endocrinologist from New York in 1931. It is responsible for significant and psychological disorders which justify the study of its pathogenesis. Hormonal dysfunction has been demonstrated among women who are at risk for PMS; nevertheless, it has been shown that neurological transducers are also affected, such as GABAergic, serotoninergic and endorphinic systems. Interactions between the two systems allow to raise the hypothesis of an inbalance between GABAergic and progesterone derived neurosteroids in a psychoneuroendocrinological model. Based on this hypothesis, psychological symptoms can be efficiently treated by anxiolytic or antidepressant treatment. On the other hand, progesterone derivatives and, sometimes, diuretics, are useful on physical symptoms. As far as we know there is so far no single treatment of demonstrated efficacy in the PMS. PMID- 10389470 TI - [Hyperandrogenism: clinical aspects, investigation and treatment]. AB - Androgen excess (AE) is one of the most common endocrine disorders, affecting 10% of adult women before the menopause. The clinical picture varies widely depending on the etiology of AE. Most of these women are suffering from hirsutism, acne, menstrual disturbances, anovulation and obesity. Virilization is unusual, except in patients with ovary or adrenal cancer. Polycystic ovary syndrome (PCOS) and idiopathic hirsutism (IH) are the most frequent causes of androgen excess, accounting for more than 90% of the cases. The pathogenesis of PCOS is still an unresolved problem. A hereditary predisposition has been suggested. Enzymatic deficiency is a less frequent cause of AE, the most common deficiency being the non classic 21-OH deficiency (NCAH). AE has been implicated as a side effect of many drugs. Ovary and adrenal tumours are unusual, however, they must be considered especially in case of severe hirsutism or virilization. Complementary investigations are selected based on the result of clinical examination. Pharmacologic therapy, usually with anti-androgens, is the most widely used treatment for PCOS, IH and NCAH. Surgical therapy should be considered only when there is a particular indication such as Cushing's syndrome, ovary or adrenal tumours. PMID- 10389471 TI - [Genital warts]. AB - HPV related pathologies are the commonest reasons for consulting a gynaecologist. Genital warts are associated with infection by HPV type 6 or 11. Etiopathogenetic factors and treatment modalities are reviewed. PMID- 10389472 TI - [Meno-metrorrhagia]. AB - Menometrorrhagia is frequent. It consists in menorrhagia (excessive menstrual flow and duration) and metrorrhagia (irregular, excessive flow and duration). Three different types of aetiology occur: general extra-gynaecological causes, endocrine causes and organic causes. This last group is made of myomas, polyps, endometrial hyperplasia, adenomyosis and uterine cancers. Dysfunctional uterine bleedings do not find their cause in one of these three main causes. The diagnosis is based on three types of complementary investigations: endo-uterine cytological and histological samplings, medical imaging of which endovaginal echography is the most accurate, and diagnostic hysteroscopy. This triad allows to reach a very precise diagnosis in order to exclude a malignant lesion. Thanks to this precise diagnosis, the therapeutic decision is made according to the nature of the lesion to be treated, the desire to retain fertility, and age. Medical and surgical treatments are possible. In most cases of general extra gynaecological and endocrine causes, medical treatment is efficient and etiological. When organic uterine lesions are present, several medical treatments are efficient by suppressing the cause of bleeding or by symptomatic action. Main medical treatments are: anti-fibrinolytic agents, nonsteroidal anti-inflammatory drugs, progestin, oral contraceptive pills, GnRH agonists and danazol. The surgical treatment consists in endoscopic techniques (operative hysteroscopy and laparoscopy) and hysterectomy performed by vaginal route with or without laparoscopic preparation, by laparoscopic approach only or by classical laparotomy. Currently, the classical D & C has become essentially a diagnostic method. Surgical treatment is necessary after failure of a medical treatment or in the presence of a lesion not directly accessible to medical therapy. The efficacy of conservative endoscopic techniques depends on the respect of the indications of these techniques. These allow to reduce the number of hysterectomies for benign lesions by up to 50%. PMID- 10389474 TI - [Cervical pap smears: techniques and clinical significance]. AB - To be satisfactory, a cervical pap smear must be correctly fixed, and must contain endocervical cylindric cells and/or squamous metaplasic cells and mucus. The cytologist's conclusions and his recommendations must be expressed clearly. Any atypical findings must be controlled. If they persist, the patient will be oriented toward a colposcopy examination where targeted biopsies will be performed. PMID- 10389473 TI - [Vaginal infections and sexually transmitted diseases]. AB - The diagnosis and treatment of vaginal and sexually transmitted infections constitute an important part of the activity of the gynaecologist and of the general practitioner. In this review article, we will describe various clinical entities, including bacterial vaginosis, vulvovaginal Candidiasis, trichomoniasis, gonorrhea, syphilis, genital herpes, Chlamydial infection, and pelvic inflammatory disease. The acquired immunodeficiency syndrome will not be described here. PMID- 10389475 TI - [Mammary pathology]. AB - Mammary pathology is dominated by breast cancer. We will successively review various therapeutic consensus, high risk factors, bordeline lesions and carcinomas in situ. PMID- 10389476 TI - [Some concerns and semantic confusions about feminine skin]. AB - Some characteristics of the skin are badly perceived by women living in a society where youth and idealized physical aspect are respected as a cult. Among the most frequent concerns, one should cite dry skin, wrinkles, striae distensae, cellulite, diffuse hair loss, hyperpilosity, as well as the unwanted effects of menopause and of the immoderate exposure to sun and sunbeds. Most of these concerns are subject to semantic confusions sustained by some medias. They are frequently targets of a worldwide ballyhoo conveyed to the general public without the support of sound physiopathological basis. PMID- 10389478 TI - [Depression in women]. AB - The woman is twice more likely than man to suffer from depression. The reasons can be biological, psychological and social. The woman is particularly sensitive to depression during her reproduction period depending on the psychological modifications she faces and the hormone variations she has to go through. The general practitioner can encounter various depressive disorders with the following specific clinical and therapeutic aspects: premenstrual syndrome, gravidic depression, postpartum disorders, menopause depression. PMID- 10389477 TI - [Perimenstrual dermatoses: a comment fact of chronobiology]. AB - More than 15 common or specific dermatoses may be worsened during the premenstrual phase. Acne, herpes simplex and the autoimmune progesterone dermatitis represent typical examples. PMID- 10389479 TI - [Specific phobias in women]. AB - Specific phobias are common, leading to a high level of suffering and disability when subjects have to face the phobic stimulus. According to the epidemiologic survey of Liege, one fifth of the population exhibits some phobic disorder during lifetime. Women are more affected than men (2:1). This article reviews definitions, etiopathogeny and treatments of specific phobias, and tries to explain the reasons why the trouble is more frequent in women. PMID- 10389480 TI - [Postmenopausal osteoporosis in women]. AB - Postmenopausal osteoporosis is now considered a major public health problem in aging women, due to the burden related to the consequent fractures. Over recent years, several pharmacological approaches were developed for the prevention and treatment of osteoporosis. Besides regular physical exercise and calcium rich diet, calcium supplementation can be suggested to both genders, after seventy years as well as systematic vitamin D supplementation in order to cope with the frequent lack observed in our country. Hormone replacement therapy is the first choice in prevention of postmenopausal osteoporosis. Based on a careful evaluation of the needs of a postmenopausal woman as well as on the risk/benefit ratio derived from her individual risk factors, selective estrogen receptor modulators (raloxifene) or second generation bisphosphonates (alendronate) can be considered as alternative to estrogens. Due to its prohibitive cost, nasal calcitonin should be only considered for very specific cases. In osteoporosis confirmed by bone densitometry or by occurrence of fractures, bisphosphonate (alendronate) reduces subsequent fracture rate. Fluoride salts can, in some cases, improve spinal symptomatic osteoporosis. The use of etidronate, a molecule from the past, should be avoided as much as possible and, at any rate, strictly restricted to its legal indication including women with several vertebral crush fractures and severely decreased bone mineral density. PMID- 10389481 TI - [Urinary incontinence in elderly women]. AB - Physical, psychological and economical consequences of urinary incontinence of the elderly woman are underestimated. It often results in depression, social isolation and early institutionalisation. It is often the key factor that determines the decision of institutionalisation, which represent the most important part of the total cost of urinary incontinence. This problem is too often neglected and deserves considerable attention. PMID- 10389482 TI - [Aesthetic surgery update]. AB - Aesthetic surgery has its origin in plastic surgery. It has considerably developed to such a point that the number of procedures performed at the present time is well over the reconstructive procedures. After having emphasized the importance of patient's motivation and the high level of specific technical training of the plastic surgeon, the most common procedures are illustrated. Their major advantages and pitfalls are outlined. PMID- 10389483 TI - [Possibilities for computer tomography of the equine head based on two cases with a fracture of the base of the skull]. AB - For the past 2 years computed tomography is used at the Veterinary School, University of Zurich. This new imaging modality enables the detection of abnormalities occurring in small and large animals which were previously not visible with imaging techniques. Subjects of this study were a foal and a small pony, both with suspected head trauma. Routine radiography could not explain any of the neurological deficiencies. In the first case a basilar skull fracture along with a focal brain hemorrhage was detected, in the second case multiple basilar skull fractures were seen. The computed tomographic examination after intravenous injection of an iodine-containing contrast medium also revealed epidural hemorrhage. These two cases demonstrate the diagnostic use of computed tomography as an extremely valuable asset when diagnosing head trauma in horses. PMID- 10389484 TI - Abortion rate during the second half of gestation of cows undergoing surgery in lateral recumbency as compared with cows of a control population. AB - In a retrospective study, we compared the abortion rates of 59 cows in the second half of gestation, admitted to the Clinic for Food Animals and Horses (1993-1996) with a strictly focal problem (head region, udder and teats, locomotor system) requiring surgery in lateral recumbency, to the abortion rate of a control cow population. Before surgery, a uterus relaxant was given in 42 cases, and 17 cows were untreated as to pregnancy. Cows included in the reproduction health program of the Department of Fertility and Reproduction were used as a control population. Data of 1,736 cows pregnant in the second half of gestation per year were available for the same time period (1993-1996). Abortion rate was 5.08% for the cows undergoing surgery in lateral recumbency as compared with 5.18% for the cows of the control population. There was no significant difference between the two populations (P > 0.05). Thus, the risk for abortion in the cows in question is not elevated as compared with the control population. PMID- 10389485 TI - [Mitotane treatment in a dog with a recurring adrenocortical carcinoma--a case report]. AB - In a 7 year old female poodle an adrenocortical tumor was diagnosed on basis of laboratory and ultrasonographic examinations. One year after adrenalectomy, a relapse was diagnosed, at that time the suspicion of metastases in the liver arose for the first time. By treatment with Mitotane in a dose aiming at completely destroying the adrenal cortex, a complete disappearance of the tumor as well as a dramatic reduction of the size of the metastases could be achieved. 12 months after the begin of the chemotherapy, the dog is in good general condition. PMID- 10389486 TI - [Resistance situation and enterotoxin production capacity of Staphylococcus aureus strains from bovine mastitis milk samples]. AB - In total, 63 S. aureus strains from mastitis milk samples of different animals in 57 farms were isolated. In 14 (22%) of the S. aureus strains resistancies against one or several of the examined antibiotics could be observed whereby six resistance patterns were found. 14.3% of the strains were penicillin resistant. 34 (54%) of the 63 S. aureus produced enterotoxins (SE). Three strains formed SEA, 21 SEC, three SED and seven strains 2 SE, SEAC, SEAD or SEBD. PMID- 10389487 TI - Cortical populations and behaviour: Hebb's thread. AB - This paper discusses work on the function of the motor cortex as revealed by single cell recordings in monkeys and artificial neural network modelling. Our key conceptual approach both in behavioural neuroscience and neural network modeling of motor cortical function relies on reconstructing, visualizing, and modelling the activity in neuronal populations, indeed a key concept advanced by Hebb (1949). The behaviour investigated ranges from exertion of isometric force to pointing movements to complex cognitive processing. The functional properties of single cells with respect to the direction of movement in space are described as well as a population code which provides a unique measure for this direction. Finally, the results of modeling studies are discussed in which directional population activity is used as an input to an artificial neural network to drive a simulated arm. PMID- 10389489 TI - Neuromodulation, development and synaptic plasticity. AB - We discuss parallels in the mechanisms underlying use-dependent synaptic plasticity during development and long-term potentiation (LTP) and long-term depression (LTD) in neocortical synapses. Neuromodulators, such as norepinephrine, serotonin, and acetylcholine have also been implicated in regulating both developmental plasticity and LTP/LTD. There are many potential levels of interaction between neuromodulators and plasticity. Ion channels are substrates for modulation in many cell types. We discuss examples of modulation of voltage-gated Ca2+ channels and Ca(2+)-dependent K+ channels and the consequences for neocortical pyramidal cell firing behaviour. At the time when developmental plasticity is most evident in rat cortex, the substrate for modulation is changing as the densities and relative proportions of various ion channels types are altered during ontogeny. We discuss examples of changes in K+ and Ca2+ channels and the consequence for modulation of neuronal activity. PMID- 10389488 TI - Behavioural context and a distributed system: metabolic mapping studies of the basal ganglia. AB - Behavioural context is known to affect neural activity in the striatum. Responses of single cells increase to rewarding stimuli, or drop out as a bar press or saccade is learned. Networks that can accomplish a unique response to changing contexts are of particular interest to systems neuroscience and were a part of Hebb's interest in perception and learning. An overall map of the striatum that localizes changes related to this remarkable phenomenon of contextual responses contributes to our understanding of anatomical substrates of neural systems that integrate information, and may lead us to new striatal regions to study synaptic mechanisms of learning. PMID- 10389490 TI - Synaptic plasticity and the organization of behaviour after early and late brain injury. AB - Hebb proposed that synaptic change underlies behavioural and cognitive plasticity. When applied to recovery from brain injury, the general hypothesis is that if there is recovery following brain injury, then there ought to be a correlated synaptic change, which is presumed to be responsible for recovery. In contrast, if recovery fails to occur, or expected recovery is blocked in some manner, then the synaptic change will likely not be present. Systematic study of functional recovery and synaptic change following brain injury at different ages supports these predictions. Good recovery is always correlated with enhanced connectivity whereas poor recovery is always correlated with an absence of reorganized connectivity. Furthermore, factors that stimulate recovery, such as neurotrophins or experience, stimulate synaptic change and functional recovery. Factors that retard recovery, such as depletion of neuromodulators, also block synaptic change. These results thus support Hebb's general idea that synaptic plasticity is related to behavioural change. PMID- 10389491 TI - Ontogenetic constraints on neural and behavioral plasticity: evidence from imprinting and face processing. AB - This paper addresses the extent and limits on brain plasticity during development through the detailed study of imprinting in the domestic chick and the development of face processing in human infants. In both of these systems, evidence for constraints on plasticity is reviewed. The first source of constraint comes from the basic architecture of learning mechanisms that support plasticity. With regard to the chick, a specific "Hebbian" model based on the known neural circuitry of the region of the brain involved is presented and discussed. In human infants, a more abstract model inspired by cortical circuitry is mentioned. The second source of constraint comes from biases on the nature of the stimuli selected for attention by the young organism. Both in the chick and the human there is evidence for a subcortical brain system which orients their attention toward conspecifics, and particularly to their faces. It is argued that these systems tutor, or bias the input to, the more plastic learning systems. PMID- 10389492 TI - The first panic attack: a neurobiological theory. AB - We extend a neurodevelopmental model of specific phobias to the etiology of an initial panic attack and its elaboration into panic disorder. An important difference between the initial panic attack and specific phobia is the developmental timing of critical emotional experience: Those occurring early in development lead to panic; those occurring later in development lead to specific phobia. By this account, sensory and emotional experiences that occur early in development are stored in a set of modules, each with a unique developmental trajectory. Reinstatement, which occurs during hormonal stress, produces an aggregate of sensory and emotional memories and the first experience of an unexplained panic attack. Panic disorder, which evolves from unexplained panic attacks, involves retrieval of a disaggregate set of sensory and emotional memory fragments supplemented by an inferential fitting of an explanatory context to this incomplete aggregate. PMID- 10389493 TI - The deterioration of semantic memory in Alzheimer's disease. AB - Semantic memory impairment is a common feature of dementia of the Alzheimer type (DAT). Recent research has shown that patients with DAT are more impaired (relative to non-demented controls) in generating exemplars from a particular semantic category (e.g., animals) than words beginning with a particular letter, exhibit an altered temporal dynamic during the production of category exemplars, are impaired on confrontation naming tasks and make predominantly superordinate or semantically related errors, consistently misidentify the same objects across a variety of semantic tasks, and have alterations in multi-dimensional scaling models of their semantic network that are indicative of a loss of concepts and associations. These results are consistent with the view that Alzheimer's disease results in a breakdown in the organization and structure of semantic knowledge as neurodegeneration spreads to the association cortices that presumably store semantic representations. PMID- 10389494 TI - Webs, cell assemblies, and chunking in neural nets: introduction. AB - This introduction to Wickelgren (1992), describes a theory of idea representation and learning in the cerebral cortex and seven properties of Hebb's (1949) formulation of cell assemblies that have played a major role in all such neural net models. Ideas are represented in the cerebral cortex by webs (innate cell assemblies), using sparse coding with sparse, all-or-none, innate linking. Recruiting a web to represent a new idea is called chunking. The innate links that bind the neurons of a web are basal dendritic synapses. Learning modifies the apical dendritic synapses that associate neurons in one web to neurons in another web. PMID- 10389495 TI - The role of peripheral and central visual information for the directional control of manual aiming movements. AB - Seeing one's hand in visual periphery has been shown to optimize the directional accuracy of a sweeping hand movement, which is consistent with Paillard's (1980; Paillard & Amblard, 1985) two-channels model of visual information processing. However, contrary to this model, seeing one's hand in central vision, even for a brief period of time, also resulted in optimal directional accuracy. One goal of the present study was to test two opposing hypotheses proposed to explain the latter finding. As a second goal, we wanted to determine whether additional support could be found for the existence of a visual kinetic channel. The results indicated that seeing one's hand in central vision, even for a very short delay, resulted in the same accuracy as being permitted to see one's hand for the duration of the whole movement. This suggests that seeing one's hand around the target might enable one to code its location and that of the target within a single frame of reference and, thus, facilitate movement planning. In addition, the results of the present study indicated that seeing one's hand in motion while in visual periphery permitted a better directional accuracy than when this information was not available. This suggests that the movement vector, which is planned prior to movement initiation, can be quickly updated following movement initiation. PMID- 10389496 TI - Adolescent girls: an emerging priority. PMID- 10389497 TI - Influence of recall period on estimates of diarrhoea morbidity in infants in rural Tamil Nadu. AB - Data collected on 689 infants, in a study to assess the incidence of diarrhoea during infancy, is used here to quantify the extent of under-reporting in diarrhoea morbidity surveys. The study area consisted of two contiguous primary health centres in Villupuram health unit district in Tamil Nadu, south India. Each day of infancy was assigned a recall period and proportion of diarrhoeal days for various recall period computed. The proportion of diarrhoea was 11.3%, 12.0% and 11.2% for zero, one and two days of recall period, respectively, (mean 11.5%) after which the proportion decreased. The under-reporting of diarrhoea was approximately 15%, 26% and 45% with three, six and 7-13 days of recall period, respectively compared to mean. As there is considerable under-reporting of diarrhoea morbidity when recall period exceeds three days, it would be better to collect information on diarrhoea twice a week in diarrhoeal morbidity surveys. PMID- 10389498 TI - A study of health and health related social problems in the geriatric population in a rural area of Tamil Nadu. AB - About 200 individuals aged 60 years and above, out of 317 total geriatric population of a small village in Tamil Nadu were subjected for complete clinical examinations with the objective of finding major health problems and associated social problems prevalent among these study population. Main causes of illness were found to be arthritis, cataract, bronchitis, skin diseases and malnutrition. PMID- 10389499 TI - Early neonatal morbidity and mortality in a city based medical college nursery. AB - Early neonatal morbidity & mortality were studied in 7972 viable live born babies over one year period in Medical College, Calcutta. Early neonatal morbidity and mortality rate were 66.85 & 32.86 respectively. About 48% of Early neonatal deaths occurred in 1st 48 hours & 80% within first 72 hours of life. Early neonatal mortality rate per 1000 was maximum in babies born of a primi (43.03) and grand multiparous mother (103.89); maternal age less than 20 yrs. (44.15), and more than 30 yrs. (46.04) & in multiple pregnancy (142.85). It was highest in breech delivery (114.28) & with maternal disease like dribling (179), hypertension (146) & APH (116). Birth asphyxia contributed 65.26% of early neonatal deaths, followed by septicaemia (10.3%). Klebsiella was the most common (55%) organism. PMID- 10389500 TI - Impact of hospitalization on patients and their families. PMID- 10389501 TI - SPM do ORT in Uttar Pradesh. PMID- 10389502 TI - Influence of cigarette smoking on Vitamin C, glutathione and lipid peroxidation status. AB - There has been a growing interest during recent years in the role of free radicals and lipid-peroxidation at tissue-level for the causation of cancer and other age-related diseases like atherosclerosis, rheumatoid arthritis, cataract etc. Free radicals and increased lipid peroxidation play a significant role for causation of human diseases by oxidative damage and functional degeneration of the tissues. Vitamin C, a well-known dietary antioxidant, and other enzymatic antioxidants like glutathione can protect the lipids of lipoproteins and other biomembranes against peroxidative damage by intercepting oxidants before they can attack the tissues. But cigarette smoking was found to affect the antioxidant protective action of Vitamin C, glutathione etc. A group of adult male smokers in this study were found to have lowered Vitamin 'C' & glutathione levels, but increased lipid-peroxide levels in their blood. Thus the increased pathogenicity of the smoking may also be due to indirect biochemical effect of enhanced oxidative stress by increased lipid-peroxidation and lowered Vitamin C & other antioxidants at tissue-level. PMID- 10389503 TI - Reinvasion of Calcutta city by Aedes albopictus: the proven vector of dengue in suburban areas. PMID- 10389504 TI - Safe motherhood: initiatives to make it safer. PMID- 10389505 TI - Industrial ocular morbidity in a north Indian town. AB - A study on industrial ocular morbidity was carried out in 6 industrial establishments at Saharanpur. The mean age of the respondents was 35.3 years. 58.2% were regular floor staff. 10.6% professed suffering from an industrial ocular injury. 60% of these injuries were sustained by ocular metallic trauma. 51.9% complained of ocular symptoms at the time of the survey. The frequency of ocular complaints increased with age. The point prevalence of ocular morbidity was 746.03/1000 industrial workers. Refractive errors were the commonest ocular condition (56.7%) observed, followed by Trachoma (32.6%). The highest prevalence of morbidity was recorded among workers above 44 years. Clerical and managerial personnel had higher prevalence compared to other jobs. Only 3.6% of the floor workers were using protective devices while on the job. PMID- 10389506 TI - A pilot study on neuroepidemiology in urban Bengal. AB - A pilot survey was undertaken in an urban district population of West Bengal, to study the magnitude of problems of Neurological disorders in the Community. The study was performed by Professionals, headed by a Neurologist, using accepted WHO protocol (1981) for neuroepidemiological survey. Point prevalence rate was found to be 75 per 1000. Sensitivity and specificity of the case study came out to be 98.97% and 99.6% respectively. PMID- 10389507 TI - Screening for obesity in affluent females: body mass index and its comparison with skin fold thickness. AB - In the present study 625 females above 15 years of age residing in affluent localities of Varanasi city were selected by multi-stage stratified random sampling technique. Body Mass Index (BMI) and Skin-Fold Thickness (SFT) were compared as indices of obesity. Prevalence of obesity by BMI and SFT was 30.24 and 49.12 respectively. SFT gave significantly higher prevalence rate of obesity as compared to BMI. It is possible that western population based SFT cut-off points may not be truly applicable to Indian study. The sensitivity, specificity and predictive value of 'sum of SFT at four sites' were calculated at different cut-off points, and it was observed, that values > or = 90 mm is the best cut-off point instead of 80 mm, for detecting obesity in the Indian context. PMID- 10389508 TI - Risk factors and protective factors of pelvic inflammatory disease: a case control study. AB - Risk factors and some protective factors for pelvic inflammatory disease (PID) in women were assessed in a case control study. Use of intrauterine device (OR = 3.98, p < 0.0001), sexual activity with multiple partners and younger age (ORs = 3.97, 1.9 and p = 0.0003, 0.0034, respectively), history of previous PID (OR = 4.08, p = 0.004) and history of minor gynecologic operation (OR = 3.07, p = 0.0158) were significant risk factors for PID. Pregnancy was a significant protective factor (OR = 0.25, p = .0074). Sterilisation had a significant protective effect (OR = 0.37, p = 0.0443) on multivariate analysis but not significant on univariate analysis. The results indicated that, almost half of the PID load on the population can be reduced by proper handling of four risk factors namely, use of IUD, sexual activity with multiple partners, history of previous PID and history of minor gynecologic operation (PARP = 0.2146, 0.1101, 0.0824 and 0.0794, respectively). PMID- 10389509 TI - Behavioural factors of Anopheles stephensi--the principal vector of malaria in Calcutta city. PMID- 10389510 TI - Resurgence of malaria in Calcutta in 1995: a hospital based study. AB - A total of 405 cases of fever who were either admitted to the Hospital or attended in paediatric out patient Department or Emergency of Medical College Hospital, Calcutta between January '95 and November '95 were included in the study. Majority of cases presented with usual features of malaria like fever with chill and rigor, hepatosplenomegaly, pallor. Apart from these, complicated manifestations like shock, convulsion D.I.C and jaundice were also observed. Some unusual presentations with severe diarrhoea, dehydration and features like that of acute viral respiratory tract infection were highly confusing in terms of clinical diagnosis. P. falciparum was observed in 35.5% of cases. Overall therapeutic response to chloroquin was good, However, two patients died of cerebral Malaria. Five cases of severe malaria were caused by P. vivax however, other etiological features could not be found to attribute the severe nature of these illnesses. PMID- 10389511 TI - Practice of self medication among slum-dwellers. AB - A cross sectional study including 110 households covering 630 individuals was carried out in an urban slum community to study the prevalence of practice of self medication by house to house survey. Self medication was practiced by 34.55% respondents and prevalent among all the age groups. Allopathic drugs were commonly used (78.95%). Economic inability in utilizing established medical facilities was the commonest reason for self medication (60.53%). The practice was more prevalent amongst literate but lower soci-economic class. PMID- 10389512 TI - Incidence of hepatitis B virus (HBV) infection amongst clinically diagnosed acute viral hepatitis cases and relative risk of development of HBV infection in high risk groups in Calcutta. AB - The present study revealed that 30.5% of acute infective hepatitis were due to the infection of Hepatitis B virus (HBV) however, 8% controls also showed HBV positivity. The possible route of infection of HBV in our country were Parenteral in 51.9%, Sexual in 24% and Unidentified in 24.1% cases. HBV marker positivity was 45.5% amongst health care workers 33.3% in recipients of multiple blood and blood product transfusion, 25% in sexual partners and their children, 20% in S.T.D. clinic attendants and 10% in patients on haemodialysis. PMID- 10389513 TI - Reproductive and child health programme. PMID- 10389514 TI - Presidential address. IPHA. Indian Public Health Association. PMID- 10389515 TI - Dr B. C. Dasgupta Memorial Oration. Rural health services and human resources development in India--an overview. PMID- 10389516 TI - K. N. Rao Memorial Oration. Liberalization of the national economy-agenda for health care policy and administration. PMID- 10389517 TI - Dr. P. C. Sen Memorial Award Paper. Status of salt iodisation and iodine deficiency in selected districts of different states of India. AB - Iodine deficiency disorders (IDD) is a major public health problem. Surveys conducted by the National Goitre Survey team of the Directorate General of Health Services during the past three decades have revealed a high prevalence of endemic goitre in different states. Out of a total of 267 districts surveyed till date, 226 have been reported to be endemic to iodine deficiency. A successful measure for the prevention of IDD is salt iodisation. The Salt department, Government of India has taken an intensive programme of production of iodised salt in the country. The production has increased from 1.5 lakh metric tonnes in 1984 to 40 lakh metric tonnes in 1996. To assess the impact of increased production of iodised salt on the availability of iodised salt at the beneficiary and trader level and also on the status of iodine deficiency, surveys were undertaken in selected districts of 10 states and 2 union territories of the country. These studies have been presented and discussed here. PMID- 10389518 TI - Dr. P. C. Sen Memorial Award Paper. A study of HIV infection in thalassaemia patients of rural Bengal. AB - Thalassaemia patients receiving repeated blood transfusions are vulnerable to transfusion related infections. HIV infection is the most life threatening of them all. Blood being the most efficient mode of transmission of HIV, increases the risk of infection even further. Although the National AIDS Control programme has laid down stringent rules regarding blood safety, it remained to be seen whether they were being followed meticulously especially in rural areas. The present study was conducted to identify the HIV status of multi-transfused thalassaemia patients attending hospital blood banks of rural Bengal. Only 3 (0.9%) of the 330 thalassaemia patients examined were found to be HIV positive. Although the situation has not reached alarming proportions, yet appropriate control measures must be adopted on a mass scale to prevent further spread of the world wide pandemic. PMID- 10389519 TI - City health dilemma on pavement dwellers. PMID- 10389520 TI - Epidemiology of peptic ulcer in north Bengal, India. AB - A retrospective analysis of hospital records of inpatients of Medicine department of North Bengal Medical College and Hospital during the period between 1988-90 revealed that 6.2% of all medical admissions were on account of peptic ulcer and or acute gastritis. Duodenal ulcer was prevalent accounting for more than 50% of the cases of peptic ulcer and acute gastritis. Duodenal ulcer was more common in the working age groups (> 21 years) among males and more than 31 yrs among females. Gastric ulcer was more common among older age groups. In respect to ethnicity, Bengali speaking hindus showed high probability for gastric ulcers in both sexes. The probability for duodenal ulcer was within confidence limits among all social groups. PMID- 10389521 TI - Bancroftian filariasis in Namrup tea estate, district Dibrugarh, Assam. AB - Filariasis survey in a randomly selected tea estate of district Dibrugrah revealed 6.7% infection of Wuchereria bancrofti in labour population with microfilaria (mf) rate of 7.6% in males and 5.9% in females. The mf rate increased progressively with the age which however, dropped in 31-40 age group of males and in 41-50 age group of females. Chronic filariasis diseases rate was 2.7%. The involvement of genitals in manifesting chronic filariasis was significantly higher than of the lower extremities. Infection and infectivity rates in the vector mosquito, Culex quinquefasciatus were 6.1% and 4.6% respectively with mean L3 load per infective mosquito of 8.5. Drains, land, peridomestic ditches were chief breeding habitats of Cules quinquefasciatus in the tea estate. PMID- 10389522 TI - Hepatitis D infectivity profile among hepatitis B infected hospitalised patients in Calcutta. AB - 450 hospitalised cases of hepatic and non hepatic disorders and 100 normal individuals were examined for serum Hepatitis B Surface antigen and Delta Virus antigen by ELISA to find out its association with different clinical disorders. 105 patients (23.3%) and 2 control (2%) were positive for HBsAG. 60 cases with jaundice (26%) were HBsAg positive. 65% of HBsAg positive jaundiced patients had serum bilirubin level more than 2 mg per dl with a mean SGPT level of 488 iu/L. Only two cases were positive for HDV antigen among 60 HBsAg positive jaundice patients indicating a lower rate of prevalence of infection (3.3%). 62 (59%) out of 105 HBsAg positive cases did not show any history of blood transfusion or surgical interference indicating a positive HBV transmission through needle prick during investigative procedures. PMID- 10389523 TI - Comparative efficacy of three measles vaccines in Indian children. AB - A Comparative study of three types of measles vaccines was undertaken among 1005 children. Of these 527 were vaccinated with the Serum Institute of India (SII) vaccine, 230 with Schwarz (SC) and 248 were vaccinated with Edmonston-Zegreb (EZ) vaccine (imported from Zegreb). Though the majority of children reacted favourably with all the three vaccines (SII: 98.43%; SC: 93.40%; EZ: 93.0%) with a rise in titre, but the percentage of seroconversion was significantly higher with the SII vaccine (p < 0.01). The Schwarz and Edmonston Zagreb vaccines showed significantly less GM titre as compared with the other age group i.e. 9-12 months (p < 0.05). With Serum Institute of India (SII) vaccine the GM titres were almost similar in the different age groups. The overall GM titre obtained with the SII vaccine was significantly higher than the SC vaccine (p > 0.001) as well as the EZ vaccine (p > 0.001). It is of interest to note that among the infants, 22.5% children had measles antibody in them before vaccination. PMID- 10389525 TI - Drug resistance to the first line of antitubercular regimen (a preliminary report). AB - Sputum samples from 100 patients of pulmonary tuberculosis were processed. These patients were admitted in group of Tuberculosis Hospital at Sewri, Mumbai, which is a referral tuberculosis hospital. Isolates were identified as M. Tuberculosis by biochemical tests. Antitubercular sensitivity testing for Isoniazid, Rifampicin, Ethambutol and Streptomycin was performed by resistance ratio method. Total resistance was 36% in our study. Resistance to Isoniazid was 61%: to Rifampicin was 50%, to Ethambutol was 8% and to Streptomycin was 41%. Primary drug resistance to Isoniazid was 45% to Rifampicin was 27%, to Ethambutol was 9%, and to Streptomycin was 54%. Secondary drug resistance to Isoniazid was 68% to Rifampicin was 60%, to Ethambutol was 8% and to Streptomycin was 36%. Secondary drug resistance to Isoniazid and Rifampicin is rising as compared to primary drug resistance to Isoniazid and Rifampicin. This is statistically significant (p < 0.001). 11 out of 36 cases (30%) showed multi drug resistance to Isoniazid and Rifampicin. PMID- 10389524 TI - Sentinel surveillance on poliomyelitis and neonatal tetanus: a report. AB - A 5 year sentinel surveillance (1989-93) of paralytic poliomyelitis and neonatal tetanus was undertaken at a rural Medical College Hospital at Burdwan, West Bengal. Poliomyelitis incidence showed an overall declining trend, which corroborated with the increased OPV coverage over the years. Incidence was more in males than females. Male:female ratio varied between 2.31:1 to 1.2:1. From 1989 to 1992, age-shift in poliomyelitis was observed when more cases were occurring above 1 year subjects. Cases were reported to be high during the months June to September every year. Incidence of neonatal tetanus (NNT) also showed a declining trend during the 5 year study period. A male preponderance was observed. NNT cases were more prevalent during the months between August and November. In an attempt for eradication of paralytic poliomyelitis and elimination of NNT, containment and other public health measures were undertaken a part of surveillance activities. The obstacles encountered in the surveillance system, as well as lacunae identified in undertaking appropriate health measures was discussed. PMID- 10389526 TI - Sterility testing of disposable syringes and needles marketed in Calcutta. AB - Presterilized (disposable) syringes and needles were subjected to sterility testing for aerobic cultures. It was found that 56.3% of the samples were contaminated indicating failure of the sterilisation process. The implications of this could be far reaching and is discussed alongwith. PMID- 10389527 TI - Treatment seeking behaviour in sexually transmitted diseases. AB - The present study has been conducted to assess social and behavioural factors predisposing individuals suffering from sexually transmitted diseases to seek treatment and the role of the health provider in them. Out results showed that the demographic, socio-economic and behavioural characteristics of patients seeking treatment at alternative places and those attending the referral hospital in the first instance were comparable. Inhibition, time and distance were important considerations for selecting a health facility. Private clinics were the most preferred (72.4%) source of treatment. In 60.3% of cases written prescriptions were not given and advice regarding treatment of sexual partner was not there in any of the cases. 98.3% of the patients lacked awareness about their disease and 91.4% patients about the treatment they were receiving. PMID- 10389528 TI - Pre-operational health status of a rural community living around the Kakrapar Atomic Power Station. AB - Pre-operational health survey was conducted around Kakrapar Atomic Power Station (KAPS) during 1991-92, just one year before the first Unit of KAPS commenced operation on September 3, 1992. Total seventeen villages were selected around KAPS, which were divided into four clusters. Total population was 14,976, of which 7540 were male and 7436 were female. Door to door 100% survey was conducted with the help of Medical doctor. Health survey was conducted specially to find out the prevalence of radiation related diseases such as congenital deformity, mental retardation, cataract, goiter (grade II & IV), reproductive disorders, and cancer, besides study of morbidity pattern and age and sex specific deaths. Results revealed that prevalent diseases pattern fell well within the range of natural incidences of diseases of rural population. PMID- 10389529 TI - Survival of enteropathogenic Escherichia coli exposed to adverse conditions. AB - Enteropathogenic Escherichia coli (EPEC) was exposed to different adverse conditions for varying period of time to assess its survival under such circumstances. From the results it is extrapolated that EPEC survive for a very long time when shielded from sunlight and after several hours of exposure to UV irradiation. PMID- 10389530 TI - [The stumbling toddler]. AB - Four previously healthy children, two boys aged 5 and one boy and one girl aged 4 more or less acutely developed a stumbling gait. The causes varied from benign such as postviral acute cerebellar ataxia and benign paroxysmal vertigo to potentially life-threatening such as intoxication with benzodiazepines and medulloblastoma. Treatment led to complete or partial recovery. (Sub)acute balance disorders in previously healthy children can be due to cerebellar ataxia, vestibular disorders and abnormal proprioception. Ancillary investigations are warranted in case of gradually developing ataxia, accompanying neurological deficits, suspicion of intoxication, recurrent or familial ataxia, no spontaneous remission or even progression. In children with an isolated cerebellar ataxia without these features, ancillary investigations may be avoided, although in such cases careful follow-up remains necessary. PMID- 10389531 TI - [Lowering of diastolic blood pressure < or = 90 mmHg should not be attempted, except in type 2 diabetics; the 'Hypertension optimal treatment' (HOT) trial]. AB - Recently the 'Hypertension optimal treatment' (HOT) study was reported. In this study 18,790 patients with diastolic blood pressure between 100 and 115 mmHg were randomly assigned target pressures of < or = 90, < or = 85 and < or = 80 mmHg respectively, and treated with a felodipine-based antihypertensive regimen. In all three groups an impressive fall in both diastolic and systolic blood pressures, and as a consequence very few major cardiovascular events (the primary endpoint of the study) were observed, but there was no difference in endpoint scores among the three groups. Type 2 diabetic patients fared substantially better than non-diabetic patients and they are likely to profit if their diastolic pressure is decreased below 80 mmHg. In the remaining patients rigorous maintenance of present-day standards (diastolic pressure < or = 90 mmHg) is advised. The addition of 75 mg aspirin 1 dd resulted in a modest but significant reduction of major cardiovascular events, but at the cost of increased gastrointestinal bleedings. PMID- 10389532 TI - [Legionella epidemic in the Netherlands]. AB - In March 1999 an epidemic of Legionella pneumonia developed in the province of Noord Holland, the Netherlands: by the end of April 233 patients had become ill after a visit to a flower show, in 106 of whom the diagnosis of Legionella pneumonia was established, while in 48 the disease was probable and in 4 it was possible; 23 patients died, in 16 of whom the diagnosis of legionnaires' disease was established. In 37 patients Legionella pneumophila was isolated. There was a significant association with stands on the show that displayed anticondense fluids to clean glasses, steam ironing machines and whirlpools and bubble baths, making a relation to aerosol transmission of the causative agent very probable. Legionella (the same serotype) was indeed isolated from a whirlpool. As Legionella can readily be isolated from the human environment, a limited number of species can cause disease and laboratory tests are expensive and time consuming, an effective primary preventive programme is an illusion. Prevention should be aimed at monitoring the environment of susceptible hosts and tracing sources of infection of index cases. Patients with severe pneumonia requiring intensive care must be treated with antibiotics that cover Legionella, until this cause is excluded. PMID- 10389533 TI - [Glycemic regulation and management of essential hypertension in diabetics with type 2 diabetes mellitus; the 'United Kingdom prospective diabetes study' of diabetic complications]. AB - Patients with type 2 diabetes mellitus often develop micro- and macrovascular complications. In 25% of them, complications are already present at the time of diagnosis. The principal objective of the United Kingdom prospective diabetes study was to determine if good blood glucose control and adequate treatment of hypertension in patients with type 2 diabetes mellitus can prevent development of diabetes-related complications. The question was also studied if they way in which this blood glucose control was achieved and the way of treating the blood pressure affected the prognosis. Blood glucose control was found to reduce the incidence of--especially--microvascular complications. Oral hypoglycaemic agents and insulin both play an important part in achieving good control. Treatment with metformin reduced mortality due to cardiovascular disease in obese patients. Strict control of the blood pressure reduced development of micro- and macrovascular complications; the mortality from diabetes-related disorders and the numbers of patients suffering a stroke or heart failure. Non of the antihypertensive drugs used (an ACE inhibitor and a beta-blocking agent) offered any advantages over the other. PMID- 10389534 TI - [Genetics of colorectal cancer. I. Non-polyposis and polyposis forms of hereditary colorectal cancer]. AB - About 5% of colorectal cancer cases are due to an autosomal dominant genetic predisposition with high penetrance. In this condition, the patient is carrier of a pathogenic gene mutation present in all body cells which can be transmitted to descendants, a so-called germ line mutation. The mutation is usually present in a tumour suppressor gene. Three subgroups of hereditary colorectal cancer can be distinguished on the basis of the clinical characteristics: (a) syndromes without polyposis (mostly hereditary non-polyposis colorectal carcinoma; HNPCC), (b) syndromes with adenomatous polyposis (mostly familial adenomatous polyposis; FAP) and (c) syndromes with hamartomatous polyposis. Recently, the main gene defects which underlie these syndromes were identified. Consequently, it is possible in approximately half the families with HNPCC or FAP in patients with colorectal cancer to demonstrate the causative gene defect and subsequently, by blood testing of healthy relatives to determine who is and is not a carrier of this hereditary condition. Thus, preventive measures can be directed toward family members with a demonstrable high risk of large bowel cancer. PMID- 10389535 TI - [Genetics of colorectal cancer. II. Hereditary background of sporadic and familial colorectal cancer]. AB - About 15% of patients with colorectal cancer have a positive family history: 5% have hereditary colorectal cancer (hereditary non-polyposis colorectal carcinoma (HNPCC), familial adenomatous polyposis (FAP) or some other hereditary syndrome), while in 10% no clear hereditary pattern can be recognized ('familial colorectal cancer'). In sporadic and in familial intestinal cancer, a demonstrable hereditary predisposition may undoubtedly exist. HNPCC is often characterized by microsatellite instability, i.e. an increased number of short DNA sequences in the DNA indicating a disorder in DNA repair and a mutation in a DNA 'mismatch repair' (MMR) gene. Indicative of hereditary bowel cancer on the basis of such an MMR gene mutation are: (a) presence of bowel cancer in > or = 3 relatives, (b) early age at the time of the diagnosis of 'bowel cancer', (c) multiple primary bowel tumours, (d) uterine cancer in the family and (e) bowel and uterine cancer in a woman. Recent data demand a new subdivision of hereditary bowel cancer, based upon both the clinical picture and the results of DNA-tests. The genetic alterations in colonic adenomas and carcinomas are known to a large extent. In future these insights may be important in clinical practice, such as a more individual determination of the patient's prognosis and accordingly, of the treatment and follow-up. PMID- 10389536 TI - [Periodic colonoscopic examinations of persons with a positive family history for colorectal cancer. Work Group 'Hereditary non-polyposis- colon-rectum cancers']. AB - Individuals with one first-degree relative with colorectal cancer diagnosed before age 45 years and those with two first-degree relatives with colorectal cancer run a significantly increased risk (relative risk: 4-6) of developing colorectal cancer. Based on calculation of the mortality due to colorectal cancer for the age group 50-70 years (which is higher than the mortality due to breast cancer) surveillance may be justified, e.g. by colonoscopy at 5-year intervals from the age of 45-50. The total number of people in the Netherlands in this high risk group is estimated at 10,000. The authors conclude that prospective studies are needed to assess the cost-effectiveness of such a programme. PMID- 10389537 TI - [Stereotactic radiosurgery with adjusted linear accelerator for cerebral arteriovenous malformations: preliminary results in the Netherlands]. AB - OBJECTIVE: To assess the effects of stereotactic radiosurgery of a cerebral arteriovenous malformation (AVM). DESIGN: Prospective. METHOD: In November 1991 December 1995 linear acceleration radiosurgery was performed on 29 patients for their 30 cerebral AVMs in the University Hospital Vrije Universiteit, Amsterdam, the Netherlands. There were 15 females and 14 males with a mean age of 37.1 years (range: 13-58). Generally one isocentre was used and 15 Gy was given to the margins of the AVM at the 80% isodose. The mean target volume was 2.4 ml (range; 0.5-8.2). After 6 months, one year and thereafter every year, neurological and MRI-controls took place, in the outpatient ward. Angiography was performed after an average of 35 months (range: 24-70) to check if the AVM had obliterated. RESULTS: Angiographic post-treatment results were available in 27 patients and MRI information in one. Angiographic obliteration occurred in 20 patients (71%). No permanent radiation-induced neurological deficit was seen, nor did any hemorrhage occur during the interval between irradiation and obliteration. PMID- 10389538 TI - [Laparoscopic splenectomy for hematological diseases; results in 28 patients]. AB - OBJECTIVE: To evaluate the first results of laparoscopic splenectomy for haematological diseases and the learning curve. DESIGN: Retrospective. PATIENTS AND METHODS: Data of all patients who underwent a laparoscopic splenectomy in October 1994-July 1998 in the University Hospital Rotterdam, Department of surgery, the Netherlands, were collected from electronic databases. Data on postoperative complications were collected from medical records. Patients with splenomegaly (> 15 cm) were not eligible for the procedure. RESULTS: 28 patients were eligible for a laparoscopic splenectomy. The male:female ratio was 1:4. The mean age was 35 years. The indications for surgery were idiopathic thrombocytopenic purpura (ITP; n = 24), Gilbert syndrome (n = 1), spherocytosis (n = 1), thalassaemia (n = 1) and haemolytic anaemia with ITP (n = 1). Conversion to an open procedure was necessary in 5 of 28 laparoscopic splenectomies (18%). The median operating time was 172 minutes. Complications occurred in four patients: pneumonia (n = 2), bleeding (n = 1) and urosepsis (n = 1). The median hospital stay was 5 days (range: 1-18). The first 14 laparoscopic splenectomies differed from the following 14 by a higher conversion rate (p = 0.01), a longer operation time (p = 0.002) and a longer hospital stay (p = 0.004). In 23 out of 25 patients with ITP the thrombocyte count became normal. CONCLUSION: Laparoscopic splenectomy is associated with a learning curve, with a high incidence of conversion in the early procedures. It appears to be a safe and effective operation. PMID- 10389539 TI - [Chronic recurrent headache without neurological abnormalities. Practice guidelines of the Netherlands Society of Neurology]. PMID- 10389540 TI - [Chronic recurrent headache without neurological abnormalities. Practice guidelines of the Netherlands Society of Neurology]. PMID- 10389541 TI - [Chronic recurrent headache without neurological abnormalities. Practice guidelines of the Netherlands Society of Neurology]. PMID- 10389542 TI - [Good results from circumcisions of Muslim boys performed outside hospital]. PMID- 10389543 TI - [Good results from circumcision of Muslim boys performed outside the hospital]. PMID- 10389544 TI - [Management and choice of antibiotics for patients with an allergy to penicillin]. PMID- 10389545 TI - [Management and choice of antibiotics for patients with an allergy to penicillin]. PMID- 10389546 TI - Cytokines, chemokines, RANTES, and eotaxin. AB - Over the past several years, a number of cytokines with chemoattractive properties (chemokines) have been identified. These low molecular weight molecules have been shown to be important leukocyte chemical attractants to sites of inflammation and infection. Chemokines act on leukocytes through selective receptors and are now known to function also in leukocyte maturation, trafficking, and homing of these cells. RANTES and eotaxin (among other chemokines) are important chemoattractants for eosinophils. Since eosinophils seem to play a critical role in the production of allergic inflammation, an understanding of the mechanism of action of these chemokines may lead to new therapies for asthma and other allergic processes. PMID- 10389547 TI - Corticosteroid effects on cytokines and chemokines. AB - Cytokines as soluble proteins or growth factors involved in cellular interactions are major contributors to allergic and immune-mediated inflammation. Chemokines are chemoattractive cytokines subdivided into families based on cysteine residues. This review on corticosteroid effects on cytokines and chemokines will consider only a selected number of several of these proteins studied in our division and in other centers. Characteristics of several major cytokines up to IL-15, chemokine targets and the effect of corticosteroids on various cells, cytokines, and chemokines are reviewed. The effect of corticosteroid inhibitors of non-specific endothelial activation, selective activation of VCAM-1 expression, eosinophil priming and chemokine production related to allergic diseases is illustrated. The effect of nasal corticosteroids on IL-1 beta, RANTES, IL-6, and IL-8 is also discussed. PMID- 10389548 TI - Update on inhaled corticosteroids: safety, compliance, and new delivery systems. AB - Since the introduction of inhaled beclomethasone, inhaled corticosteroids have revolutionized the treatment of asthma. Inhaled corticosteroids provide the most potent and consistently effective long-term control of asthma. They have become the mainstay of therapy for chronic disease and have been recommended for asthmatics of all severities. During the past two decades, after the introduction of beclomethasone, several new inhaled corticosteroids have been introduced. With more widespread use of these agents, particularly in pediatric patients, concerns about safety have risen. These concerns emanate from the use of higher dose inhaled corticosteroids and higher potency molecules. The side effects of most concern are those that are similar to those associated with systemic corticosteroids. Recently, the U.S. Food and Drug Administration (FDA) issued a class warning of potential growth suppression in some pediatric patients using inhaled corticosteroids. Patient compliance with inhaled corticosteroid therapy is problematic. Some patients think that all inhaled corticosteroid therapy is inherently dangerous because they confuse it with systemic corticosteroid or even anabolic steroid use. Most of the products are available only as metered dose inhalers. Use of these inhalers is difficult and often poorly taught to patients. The two newest inhaled corticosteroids, budesonide and fluticasone, are available as dry powder inhalers with new delivery systems. PMID- 10389550 TI - Concurrent use of salmeterol with inhaled corticosteroids is more effective than inhaled corticosteroid dose increases. AB - This randomized, double-blind, parallel, multi-center study was designed to determine whether the addition of salmeterol to existing inhaled corticosteroid therapy provides greater therapeutic benefit than doubling the dose of inhaled corticosteroids in symptomatic patients with asthma. A total of 514 adults were randomized to either beclomethasone 168 micrograms plus salmeterol 42 micrograms twice daily or beclomethasone 336 micrograms twice daily for 24 weeks. Both treatments resulted in significantly improved symptom control and increased pulmonary function. However, beclomethasone plus salmeterol provided greater improvements than doubling the dose of beclomethasone (p < or = 0.05) in FEV1 and in daily-recorded measurements of morning (38 L/minute versus 20 L/minute after treatment with higher dose beclomethasone) and evening peak expiratory flow, asthma symptom scores, symptom-free days, supplemental albuterol use, and days and nights not requiring albuterol. There were no significant differences between treatment groups in the number of patients with abnormal response to corticotropin stimulation at Treatment Week 24. No treatment differences in asthma exacerbation and adverse event frequency rates were seen. Beclomethasone 168 micrograms plus salmeterol 42 micrograms administered twice daily was superior to beclomethasone 336 micrograms taken twice daily in patients symptomatic on beclomethasone 168 micrograms, with no added safety risks. PMID- 10389549 TI - Mometasone furoate nasal spray is rapidly effective in the treatment of seasonal allergic rhinitis in an outdoor (park), acute exposure setting. AB - The objective of this study was to determine the time to onset of symptom relief following a single dose of mometasone furoate nasal spray (MFNS) in symptomatic patients with seasonal allergic rhinitis (SAR). This was a single-center, placebo controlled, double-blind, randomized, parallel-group study with a 7-day run-in period followed by a single-dose administration of medication or placebo in an outdoor park setting. The park site provided an acute exposure to seasonal (tree and grass) pollens. Patients remained in the park of approximately 12 hours after dosing, during which time hourly assessments of SAR symptoms were recorded on a diary card. Two hundred thirty-nine patients with symptoms of SAR entered the study. Patients receiving any concurrent medication for treatment of their symptoms were excluded. Patients were randomized in a 1:1 ratio to receive treatment with either a single dose of MFNS (200 micrograms/or matching placebo nasal spray. Outcome measures included an assessment of overall therapeutic response and change from baseline in total nasal plus non-nasal sign/symptom severity score, total nasal sign/symptom severity score, and total non-nasal sign/symptom severity score. Improvement in total nasal symptom scores, total non nasal symptom scores, and total nasal plus non-nasal symptom scores were greater and more sustained in patients receiving MFNS than in patients receiving placebo. The mean decrease from baseline in total nasal plus non-nasal symptom scores was significantly greater in MFNS-dosed patients than in placebo-dosed patients at 5 hours after dosing (p < 0.01). The mean decrease from baseline in total nasal symptom scores was significantly greater in MFNS-dosed patients than in placebo dosed patients at 7 hours after dosing (p < 0.01). The between-treatment differences in total nasal plus non-nasal symptom scores and total nasal symptom scores remained significant for all subsequent hourly assessments through 12 hours post-dose. Patient assessments of overall response to therapy at end point were significantly different between treatment groups (p < 0.01) with 60.5% of MFNS-treated patients reporting complete, marked, or moderate relief compared with 46.5% of placebo-treated patients. Mometasone furoate nasal spray produces a statistically significant improvement in nasal symptom scores in patients with SAR by 7 hours after administration of a single 200 micrograms dose (100 micrograms in each nostril). PMID- 10389551 TI - A simple "step-test" protocol for identifying suspected unrecognized exercise induced asthma (EIA) in children. AB - The purpose of this study was to demonstrate that a simple submaximal "step-test" could be used as an exercise challenge to identify elementary school students with suspected but undiagnosed asthma. This article also describes a protocol for exercise testing that can be used in epidemiological evaluations. School age children grades 1-4 with suspected but undiagnosed asthma were identified by a 12 item questionnaire completed by a parent or guardian. Only students identified with suspected asthma by questionnaire were exercise challenged on a step-test it baseline spirometry was normal and there was no contraindication for intense aerobic activity. Possible asthma was defined as a 15% or greater decrease in FEV1 or a 25% or greater decrease in FEF25-75 from baseline at either 3 or 10 minutes. The exercise protocol included spirometry before and after stepping continuously for 5 minutes at an exercise intensity sufficient to maintain a heart rate between 150 and 200 beats per minute. Heart rate was continuously monitored throughout the exercise period. Testing was completed at school. No complications occurred during the exercise testing. Exercise testing was completed on 548 students with suspected undiagnosed asthma. Thirty students (6%) had exercise test changes in pulmonary function that met established criteria for suspecting asthma. A board-certified pediatric allergist/immunologist or private physician examined 26 of the 30 students with positive exercise testing. Asthma was diagnosed in 23 (88.89%) of these students. All students with impaired pulmonary function after exercise were able to return to class after a short period of observation. In conclusion, a simple, reproducible school-based exercise protocol can be used to identify students with suspected undiagnosed asthma. PMID- 10389552 TI - Desensitization to 5-fluorouracil. AB - Anaphylactic reactions to 5-fluorouracil (5-FU) are uncommon. We report a 40-year old female with adenocarcinoma of the ovary who had two reactions immediately after being infused with 5-FU. The second reaction occurred despite prophylaxis with steroids and antihistamines. The patient was positive to 5-FU on puncture skin testing even though there was no previous exposure to the drug. We successfully desensitized her to 5-FU using a continuous intravenous protocol with sequential increments in the fusion rates and drug concentrations. This desensitization method may be useful to manage systemic reactions to 5-FU and other drugs. PMID- 10389553 TI - Safety and efficacy of once-daily fexofenadine HCl in the treatment of autumn seasonal allergic rhinitis. AB - Fexofenadine HCl (Allegra, Telfast) is approved in the US for twice-daily dosing in the treatment of seasonal allergic rhinitis (SAR). A once-daily dose (already available in some countries outside the US) can improve patient compliance and health outcomes. This multicenter, placebo-controlled, 14-day US study was conducted to compare the safety and effectiveness of once-daily fexofenadine HCl with placebo in the treatment of patients with moderate to severe autumnal SAR symptoms. After a 1-week placebo lead-in, patients received 120 or 180 mg fexofenadine HCl or placebo at 8 A.M. Patients recorded SAR symptom severity scores instantaneously (for the 1 hour before medication; i.e., trough blood levels), and reflectively (for the previous 12 hours) at 8 A.M. and 8 P.M. The primary efficacy measure was change from baseline in average instantaneous 8 A.M. total symptom score (TSS, the sum of individual symptom scores excluding nasal congestion). In 861 intent-to-treat patients, both fexofenadine HCl doses provided significant (p < or = 0.05) improvement in 8 A.M. instantaneous TSS compared with placebo. Similarly, both fexofenadine doses were superior to placebo for reflective TSS assessments (p < or = 0.0012). There were no statistical differences in efficacy between the two fexofenadine doses, though the 180 mg dose showed a trend toward greater symptom relief. Incidence of adverse events was similar between fexofenadine and placebo groups (30.2% and 30.0%, respectively), with headache the most frequently reported adverse event (8.9% and 7.5%, respectively). In conclusion, once-daily fexofenadine HCl, 120 or 180 mg, is safe and effective in the treatment of autumnal SAR. PMID- 10389554 TI - Changes in respiratory mechanics during abdominal laparoscopic surgery in children. AB - We designed this study in order to measure the changes in respiratory mechanics during laparoscopic surgery in children. Ventilation parameters (Flow (Vi) and Peak Pressure (Pmax)) were measured and total respiratory system mechanics (resistance (Rrs) and compliance (Crs)) were derived using multiple linear regression analysis in 11 children aged 8 months to 11 years. The Pmax increased by 26.6% and the Rrs increased by 20.2% whilst the Crs decreased by 38.9% after pneumoperitoneum. These findings suggest that clinically important changes in respiratory mechanics occur as a result of the pneumoperitoneum produced during abdominal laparoscopic surgery. PMID- 10389555 TI - Comparison of 1% ropivacaine and a mixture of 2% lignocaine and 0.5% bupivacaine for peribulbar anaesthesia in cataract surgery. AB - The purpose of the study was to compare 1% ropivacaine and hyaluronidase 75 units/ml with a 1:1 mixture of 2% lignocaine and 0.5% bupivacaine and hyaluronidase 75 units/ml for peribulbar anaesthesia in cataract surgery. We conducted a double-blind randomized trial involving 100 patients. Group 1 received a peribulbar injection of 8 ml of 1% ropivacaine and hyaluronidase 75 units/ml. Group 2 received a peribulbar injection of 8 ml of a 1:1 mixture of 2% lignocaine and 0.5% bupivacaine and hyaluronidase 75 units/ml. Parameters measured were ocular and eyelid movement scores, time suitable for surgery, need for supplementary injections, verbal pain score and complications. No statistical differences were found between the two groups regarding any of the study parameters. Both groups had excellent surgical analgesia and akinesia. We conclude that 1% ropivacaine is a suitable agent for single injection peribulbar anaesthesia for cataract surgery. PMID- 10389556 TI - Blood flow velocity in the middle cerebral artery response to tourniquet release. AB - This study was undertaken to examine the effects of tourniquet release on middle cerebral arterial (MCA) blood flow velocity in patients undergoing general anaesthesia for orthopaedic surgery. In 30 patients (ASA physical status 1 or 2) undergoing orthopaedic procedures on the upper (Group U, n = 15) and lower (Group L, n = 15) extremities, MCA blood flow velocity was measured before and after tourniquet deflation using transcranial Doppler sonography. The MCA blood flow velocity increased after release of the intraoperative tourniquet and remained elevated for two minutes in Group U (P < 0.05) and for four minutes in Group L (P < 0.05). The increase in MCA blood flow velocity was greater in Group L than in Group U (P < 0.05). A positive linear correlation between MCA blood flow velocity and PETCO2 after tourniquet deflation was observed in both groups (P < 0.001). PMID- 10389557 TI - Does local anaesthetic affect the success rate of intravenous cannulation? AB - We aimed to assess whether subcutaneous lignocaine affects the success rate of intravenous cannulation using a randomized clinical trial. Pre-prepared cannulation packs, 50% containing local anaesthetic, were used to cannulate consecutive consenting patients presenting to the Emergency Department who required cannulation as part of their routine treatment. Doctors with less than four years postgraduate experience randomly selected a pack to perform cannulation and completed a data collection form after each cannulation. Eighty seven patients received lignocaine with 73 (83.9%) successfully cannulated on the first attempt, 79 patients were cannulated without lignocaine with 64 (81%) successfully cannulated on the first attempt. Subcutaneous lignocaine did not significantly affect the success rate of intravenous cannulation on the first attempt (P = 0.5). Subcutaneous lignocaine has been shown to significantly reduce the pain of intravenous cannulation. This study supports the use of local anaesthesia for all routine venous cannulation. PMID- 10389558 TI - Safety and efficacy of target controlled infusion (Diprifusor) vs manually controlled infusion of propofol for anaesthesia. AB - In this multi-centre, randomized trial, we compared the safety and efficacy of Diprifusor TCI with manually controlled infusion (MCI) of propofol for anaesthesia. With approval, 123 adult male and female patients were studied. Firstly, each investigator anaesthetized five patients to familiarize themselves with Diprifusor TCI. In Stage 2, 98 patients were randomized to receive propofol based anaesthesia via TCI or MCI. Adjuvant drugs, airway management and monitoring were managed at the discretion of the anaesthetist. Results are presented as mean (SD). Induction times were significantly longer [67 (32) vs 54 (17)s] and induction doses were significantly lower [14 (5) vs 16 (4) ml] in the TCI vs the MCI group. Recovery times and total doses were not significantly different. There were statistically but not clinically significant differences in mean arterial blood pressure and heart rate. Quality of anaesthesia and ease of control of anaesthesia were similar. We conclude that Diprifusor TCI and MCI are similar in terms of safety and efficacy. PMID- 10389559 TI - Auditory recall and response to command during recovery from propofol anaesthesia. AB - Most studies of awareness under general anaesthesia use the ability to respond to a verbal command as the primary measure of consciousness. The aim of this pilot study was to discover whether it was possible for subjects recovering from a propofol general anaesthetic to experience conscious awareness without the capability of responding to verbal command. Ten healthy volunteers received an intravenous propofol infusion (1500 mg/hr) until they were no longer conscious. The infusion was then stopped and they were given verbal commands interspersed with random numbers from a recorded tape until they were able to respond appropriately. Seven of the subjects were able to remember numbers corresponding to times 10 to 40 seconds before they responded to verbal command. In none of these subjects was there recall of the number 30 minutes later. We concluded that there is an ability to have conscious awareness of auditory input without necessarily being able to demonstrate this by responding to verbal command. PMID- 10389560 TI - Anaesthetic assistants: their role in the development and resolution of anaesthetic incidents. AB - Trained anaesthetic assistants are considered essential for the safe conduct of anaesthesia. Data from 5837 AIMS (Anaesthetic Incident Monitoring Study) reports were evaluated for issues concerning anaesthetic assistants in the generation and resolution of anaesthetic incidents. "Inadequate assistance" as a contributing factor was identified in 187 reports, whilst "skilled assistance" which minimized the incident was present in 808 cases. One hundred and seventy-two reports specifically commented on anaesthetic assistants in the narrative section of the AIMS form. All surgical specialties were represented. In 147 of these reports the assistant actually contributed to or failed to assist with the incident. Although the majority of outcomes from the reports were uneventful, prolonged stay, awareness and ICU admission did ensue in a small number of cases. The most common incidents were related to problems with equipment, communication and inadequate staffing levels (number and/or skill mix). Results from this study have implications for anaesthetic assistant staffing levels and the orientation of course content. PMID- 10389561 TI - Incidents in obstetric anaesthesia and analgesia: an analysis of 5000 AIMS reports. AB - We aimed to explore the first 5000 incidents reported to the Australian Incident Monitoring Study (AIMS) involving anaesthesia for obstetric patients and found 203 such incidents. Analysis and classification identified seven main incident groups; regional anaesthetic techniques (33%), anaesthetic equipment problems (13%), "wrong drug" errors (10%), other drug-related problems (16%), difficult/failed intubation (9%), problems with the endotracheal tube (9%) and other problems (10%). When compared to the incidents in the main database, obstetric cases were found to be over-represented with respect to accidental dural puncture, post dural puncture headache, failed intubation in emergency situations and the incidence of certain types of "wrong drug" error. The implications of these reports regarding safe practice of obstetric anaesthesia are discussed. PMID- 10389562 TI - Personality profiles of Australian anaesthetists. AB - Identification of personality traits in anaesthetists has potential implications for selection of trainees, assessment of coping strategies during times of stress and may have a role in the analysis of critical incidents. A 24 question postal questionnaire based on the Cattell 16PF inventory was sent to specialist anaesthetists in Australia. One hundred and sixty-seven replies were received (33% response rate). Personality traits did not differ when the anaesthetists were grouped for age, number of years qualified and country of qualification. City practitioners rated themselves more inquisitive than country practitioners did (P = 0.052). Female anaesthetists self-reported they were calm (P = 0.02), patient (P = 0.02) and tolerant (P = 0.02) more often than their male counterparts, whilst more males reported themselves as highly conscientious (P = 0.01). Although some traits were consistent, personality profiles showed significant heterogeneity. Further examination of how personality and coping mechanisms interact may be central to the management of stress and critical incident generation. PMID- 10389564 TI - Norpethidine accumulation and generalized seizure during pethidine patient controlled analgesia. AB - A 35-year-old, 47 kg female presented for elective laparatomy, adhesiolysis and ileostomy formation. Pre-existing neurological problems precluded placement of an epidural and IV PCA was used for postoperative analgesia. A patient request for pethidine was allowed. Twenty-three hours postoperatively, a brief generalized seizure occurred without adverse sequelae. This had been immediately preceded by myoclonic-type jerking. The cumulative pethidine dose was 3,000 mg and the norpethidine level was 1.8 micrograms.ml-1. Avoidance of pethidine for IV PCA where large cumulative doses are anticipated is advised. Seizures associated with pethidine/norpethidine toxicity can occur early during pethidine usage, and there is considerable variation in measured norpethidine levels. PMID- 10389563 TI - Disseminated intravascular coagulation in a patient undergoing removal of humeral head for pain relief. AB - Disseminated intravascular coagulation (DIC) was recently observed intraoperatively in a patient who required removal of her right humeral head for pain relief. Despite normal preoperative coagulation parameters, the patient developed wound oozing soon after suturing the skin. Coagulation profile revealed decreased platelets, plasma coagulation factors and fibrinogen in association with elevated fibrin degradation products. To manage the DIC, urinastatin and gabexate mesilate, along with blood component replacement, proved effective. PMID- 10389565 TI - Lower limb compartment syndrome resulting from malignant hyperthermia. AB - We report a case of compartment syndrome complicating malignant hyperthermia (MH) in a previously healthy patient. An intraoperative MH crisis responded to treatment with intravenous dantrolene. The patient subsequently developed a lower limb compartment syndrome which required fasciotomy. Recognition of the link between MH and compartment syndrome helps ensure prompt diagnosis and treatment of this rare complication of MH. PMID- 10389566 TI - Bilevel non-invasive ventilation in malignant large airways obstruction during chemotherapy and radiotherapy. AB - A case is described of acute respiratory failure secondary to variable intrathoracic large airway obstruction due to a lung neoplasm. Successful ventilation was achieved with facemask bilevel non-invasive ventilatory assistance allowing radiotherapy and chemotherapy to be undertaken. PMID- 10389567 TI - Recent experiences with hexadimethrine for neutralizing heparin after cardiopulmonary bypass. AB - Hexadimethrine bromide was used for the neutralization of heparin during cardiac surgery in the late 1950s. For some years, this institution has used it for patients who may be allergic to protamine. In view of the recent renewal of interest in hexadimethrine, we present four cases outlining its use during cardiac procedures in such patients. Other drugs for reversing the action of heparin such as heparinase or platelet factor IV are not yet widely available. PMID- 10389568 TI - Antenatal and preoperative genetic and clinical assessment in myotonic dystrophy. AB - The antenatal investigation of an obstetric patient with a history of myotonia is described. The smooth and striated muscle dysfunction in myotonic dystrophy renders these patients, as a group, liable to surgical correction and exposure to anaesthesia. A caesarean section is reported to illustrate the preferred timing of diagnosis and peripartum management. While regional anaesthesia is preferred, myotonic dystrophy is not a contraindication to general anaesthesia, provided risks are anticipated and steps taken to minimize complications. PMID- 10389569 TI - Caffeine overdose in a premature infant: clinical course and pharmacokinetics. AB - The elimination of caffeine was investigated in a 1860 g, 31 week gestation neonate, following the accidental administration of a 160 mg.kg-1 dose. The first serum concentration measured was 217.5 mg.l-1 at 36.5 h after dosing. Fitting of time-concentration data was performed using non-linear regression with MKMODEL. A first order elimination model was superior to a mixed order model. Parameter estimates were: clearance 0.01 l.h-1, volume of distribution 1.17 litres, elimination half-life 81 h. Toxic manifestations included hypertonia, sweating, tachycardia, cardiac failure, pulmonary oedema and metabolic disturbances (metabolic acidosis, hyperglycaemia and creatine kinase elevation). An unusual feature of this infant's illness course was gastric dilatation. These signs resolved by day 7 at a serum concentration of 60-70 mg.l-1. Caffeine clearance has traditionally been reported as either an absolute value or as directly proportional to body weight. The per kilogram model gives an erroneous impression that clearance is greatest in early childhood and then decreases with age until adult rates are reached in late adolescence. Age-related clearance values reported in the literature were reviewed using an allometric 3/4 power model. This size model demonstrates that clearance increases in infancy and reaches adult rates within the first three months of life. PMID- 10389570 TI - Undergraduate education in critical care. PMID- 10389571 TI - Optimal assessment of cervical spine trauma. PMID- 10389572 TI - Unsuccessful treatment of pulmonary hypertension by inhaled nitric oxide and aerosolized prostacyclin. PMID- 10389574 TI - Disconnect alarm failure. PMID- 10389573 TI - Air transport: hypobaric or hyperbaric? PMID- 10389575 TI - Empty bag syndrome re-visited. PMID- 10389576 TI - Hysteroscopy and gas embolism. PMID- 10389577 TI - Remifentanil for rapid sequence induction. PMID- 10389578 TI - CNS toxicity of ropivacaine. PMID- 10389579 TI - Prostatic adenocarcinoma following androgen deprivation therapy: the new difficulty in histologic interpretation. PMID- 10389580 TI - Variants of squamous cell carcinoma of the upper aerodigestive tract. PMID- 10389581 TI - Histopathologic spectrum of the cardiac conducting tissue in traumatic and noncardiac sudden death patients under 30 years of age: an analysis of 100 cases. PMID- 10389582 TI - Natural killer cell neoplasms. PMID- 10389583 TI - Practical lymphoma diagnosis: an approach to using the information organized in the REAL proposal. Revised European-American Lymphoid Neoplasm. PMID- 10389584 TI - Gross and histologic features of locally advanced breast cancer after neoadjuvant chemotherapy. PMID- 10389585 TI - Pathologic findings after therapeutic irradiation of mammary tissues and carcinomas. PMID- 10389586 TI - Heterotopic ossification of soft tissue: a review with emphasis on ossification within abdominal surgical scars. PMID- 10389587 TI - Observations on the histopathologic diagnosis of microinvasive carcinoma of the breast. AB - Our histopathologic criteria for diagnosing microinvasive carcinoma of the breast may be enunciated as follows: (1) cytologically malignant cells in the stroma associated with in situ carcinoma, (2) absence of basement membrane and myoepithelial cells around the invasive cells, (3) frequent accompanying stromal alterations in the form of myxomatous change and loosening of connective tissue, and (4) the frequent presence of an inflammatory cell infiltrate composed of lymphocytes and plasma cells. Most or all of these four features are present in cases of ductal microinvasive carcinoma of the breast, but the lobular type is not likely to be accompanied by stromal changes or a lymphoplasmacytic cell infiltrate. The minimum information regarding microinvasive carcinoma of the breast that should be conveyed in the final pathology report includes size as measured by the ocular micrometer or a statement that microinvasion refers to a lesion smaller than 1 mm, the number of foci of invasion, and the spatial distribution of the invasive foci. The nuclear grade of the invasive cells and the size, type, and nuclear grade of the accompanying DCIS should be specified. The status of margins, presence of vascular channel involvement (a rarity in microinvasive carcinoma of the breast), and degree of proliferative changes in adjacent nonneoplastic breast tissue should be reported. Immunostains for basement membrane and myoepithelial cells may be helpful in the diagnosis of microinvasive carcinoma of the breast. Sclerosing lesions such as radial scar and sclerosing adenosis can simulate microinvasive carcinoma of the breast, especially when the latter is associated with in situ carcinoma. Caution should be exercised in cases wherein in situ malignant cells may be dislodged by needling procedures or during dissection of the excised specimen. Cautery-induced artifacts also hinder optimal histologic assessment. In some cases, it is virtually impossible to determine if true invasion is present, and the statement "microinvasive carcinoma of the breast cannot be entirely excluded" may be employed as a last resort. We consider the latter diagnosis to be the last refuge of the diligent pathologist and do not recommend it unless all diagnostic measures, including examination of deeper levels and supplemental stains, have been exhausted. It may be necessary to seek an expert opinion in "difficult" cases, particularly in the event that therapeutic decisions are to be based on the determination of invasion. From a clinical perspective, the management of microinvasive carcinoma of the breast ought to be dictated by the individual circumstances in each case. Based on currently available data, which admittedly suffer from lack of diagnostic uniformity, the vast majority of patients with microinvasive carcinoma of the breast will be node-negative and can look forward to an excellent prognosis. It is hoped that since the UICC has adopted a previously recommended definition of microinvasive carcinoma of the breast, prospective or retrospective studies with uniform diagnostic criteria will be conducted that will enable more definitive conclusions regarding the treatment and prognosis of microinvasive carcinoma of the breast. PMID- 10389588 TI - The polymerase chain reaction in the detection of micrometastases and circulating tumor cells: present status and future potential. PMID- 10389589 TI - Genetic instability and the etiology of somatic PIG-A mutations in paroxysmal nocturnal hemoglobinuria. AB - Paroxysmal nocturnal hemoglobinuria (PNH) is a hematologic disorder characterized by acquired PIG-A gene mutations that lead to defective bioassembly of glycosylphosphatidylinositol (GPI) anchors and the absence of GPI-linked surface proteins. As the etiology of these acquired PIG-A gene mutations is unknown, we hypothesized that patients with PNH have overall genetic instability and acquire somatic mutations throughout their genome. We first analyzed microsatellite sequences and found equivalent size variation using DNA from GPI-negative granulocytes compared with the DNA of paired GPI-positive B cell lines or normal granulocytes. We next quantitated the frequency of mutations at the hypoxanthine guanine phosphoribosyl transferase (hprt) gene locus, and found 1 PNH patient with a large increase in hprt mutant frequency (256.7 x 10(-6) vs. 27.8 +/- 19.9 x 10(-6) for normal adults) that was confirmed on 4 independent blood samples. We also quantitated "illegitimate" VDJ genetic recombination events between the T cell receptor V gamma and J beta gene loci, and found a second PNH patient with a large increase (43.5 events per microgram of DNA vs. 1.3 +/- 0.8 events per microgram of DNA for normal adults), confirmed on 4 independent DNA samples. Both of these PNH patients are young females with no history of aplastic anemia. Our data show that PNH patients can have increased numbers of acquired somatic mutations in gene loci distinct from PIG-A. These data suggest that genetic instability may be associated with the development of PIG-A mutations that lead to the clinical picture of PNH. PMID- 10389590 TI - Anaphylactoid reaction to imiglucerase, but not to alglucerase, in a type I Gaucher patient. AB - Imiglucerase, the recombinantly produced enzyme, is gradually replacing the human placental derived alglucerase in the treatment of gaucher patients. We describe the first case, to the best of our knowledge, of an anaphylactoid reaction to imiglucerase in a patient who tolerated alglucerase. The patient was diagnosed at the age of 2 4/12 years with anemia and hepatosplenomegaly. Over the years he had suffered from marked splenomegaly, thrombocytopenia and recurrent bleeding episodes. At the age of 24 he started treatment with imiglucerase. After 3 months of treatment, immediately after starting an infusion, he experienced flushing, cough, tachycardia, palpitation, chest pain and excessive sweating, which reoccurred on a consecutive administration. Substitution with alglucerase was tolerated well, with only mild rash when he was premedicated with benadryl. Immediate skin tests to alglucerase, imiglucerase and gelatin were negative. IgG against alglucerase was undetectable. The in vitro mast cell degranulation test was positive for alglucerase, imiglucerase heamaccel (a gelatin based plasma substitute, which is a component of imiglucerase). This sensitivity to imiglucerase but not to alglucerase, raises the question of future treatment for this patient, since the production of alglucerase may cease, once imiglucerase production will cover the need for replacement enzyme. PMID- 10389591 TI - The observation of reactive thrombocytosis in New Zealand white rabbits in response to experimental Pasteurella multocida infection. AB - Reactive thrombocytosis is an increase in the circulating thrombocyte count secondary to a physiologic process within the body, often an infection. Reactive thrombocytosis is different than primary or essential thrombocytosis which is usually related to myeloproliferative neoplasia. Essential thrombocytosis is most common in adults, whereas reactive thrombocytosis is most frequently observed in children. Reactive thrombocytosis has been occasionally reported in cats, dogs and horses but has not been previously reported in the rabbit. Rabbits were challenged with virulent Pasteurella multocida. Hematologic, clinical, and culture assessments were performed prior to challenge, enabling each animal to serve as its own control. The questions asked were whether reactive thrombocytosis was a consistent phenomena and whether its presence and/or intensity was related to disease severity. All challenged rabbits demonstrated some degree of thrombocytosis in response to the infection, but individual rabbits were varied in their pattern of thrombocytosis. Elevations varied from intense to mild to undulating with durations of 1 to 11 days above 500 x 10(9)/L and 0 to 5 days above 650 x 10(9)/L. Correlation analysis was unable to demonstrate significant association between thrombocytosis, body temperature, leukocyte count, or the granulocyte lymphocyte ratio (all r < 0.2). No significant association between intensity of thrombocytosis and degree or type of pathologic lesions was observed. Thrombocytosis does not appear predictive of disease intensity or outcome. The data indicate that in the rabbit thrombocytosis is a consistent response to infection with P. multocida. Rabbits may serve as a model for the study of reactive thrombocytosis, in humans especially in children infected with Haemophilus sp., which are also a members of the bacterial family Pasteurellaceae. PMID- 10389592 TI - Antioxidant enzymatic activities in human blood cells after an allergic reaction to pollen or house dust mite. AB - Several diseases have been related to oxidative stress. Recently, antioxidant functions have also been linked to anti-inflammatory properties. Cell defenses against reactive oxygen species include antioxidant enzymes. We studied the enzymatic antioxidant capacity in human blood of both red blood and mononuclear cells from patients suffering from an allergic reaction to pollen or house dust mite. We determined superoxide dismutases (SODs), glutathione peroxidase (GSHPx) and catalase (CAT) activities in each cell type. We also determined the extent of thiobarbituric acid reactive substances (TBARS), in order to study the correlation between the cellular enzymatic activities, the redox status and the disease. In mononuclear cells from allergic patients, SODs and CAT activities were enhanced compared to controls. Conversely, a decrease in GSHPx activity was found. In erythrocytes, higher values for GSHPx and SODs and similar CAT activities were found in allergic patients and controls. Interestingly, CuZnSOD and MnSOD activities were enhanced in the same proportion for both, erythrocytes and mononuclear cells. TBARS were also enhanced in both types of cells. The respective enzymatic imbalances in mononuclear cells and erythrocytes, namely, GSHPx/SOD and CAT/SOD, and their consequences are discussed. To our knowledge, this is the first global study of antioxidant enzymes, including TBARS level determinations, in allergy. PMID- 10389593 TI - Anti-beta s-ribozyme reduces beta s mRNA levels in transgenic mice: potential application to the gene therapy of sickle cell anemia. AB - Our current strategy for gene therapy of sickle cell anemia involves retroviral vectors capable of transducing "designer" globin genes that code for novel anti sickling globins (while resisting digestion by a ribozyme), coupled with the expression of a hammerhead ribozyme that can selectively cleave the human beta s mRNA. In this report, we have tested in vivo an anti-beta s hammerhead ribozyme embedded within a cDNA coding for the luciferase reporter gene driven by the human beta-globin promoter and hyper-sensitive sites 3 and 4 of the locus control region. We have created mice transgenic for this luciferase-ribozyme construct and bred the ribozyme transgene into mice that were already transgenic for the human beta s gene. We then measured expression of the beta s transgene at the protein and RNA levels by HPLC and primer extension. The presence of the ribozyme was associated with a statistically significant reduction in the level of beta s mRNA in spleen stress reticulocytes (from 60.5 +/- 4.1% to 52.9 +/- 4.2%) and in the percentage of beta s globin chains in very young mice (from 44.5 +/- 0.6% to 40.8 +/- 0.7%). These results demonstrate that it is possible to decrease the concentration of beta s chains and mRNA with the help of a hammerhead ribozyme. While the enormous amount of globin mRNA in reticulocytes is a challenge for ribozyme technology, the exquisite dependence of the delay time for formation of Hb S nuclei on the concentration of Hb S in red blood cells suggests that even a modest reduction in Hb S concentration would have therapeutic value. PMID- 10389594 TI - AML-M0: a review of laboratory features and proposal of new diagnostic criteria. AB - In 1991, the FAB group published a proposal to designate acute leukemias with minimal signs of myeloid differentiation as AML-M0. This proposal was meant to offer a provisional basis for the study of immature myeloid forms, with the understanding that it was susceptible to changes and improvements with new information derived from the laboratory. Since then there have been a number of reports detailing the biological and clinical features of patients with AML-M0. In this article we review the laboratory data acquired from various sources and suggest a partial modification of diagnostic criteria. PMID- 10389595 TI - T cell numbers relate to bone involvement in Gaucher disease. AB - The major elements of bone pathology in Gaucher disease are a failure of osteoclast and osteoblast function, resulting in osteopenia and also osteonecrosis. T lymphocytes have recently been found to be involved in the regulation of osteoblast/osteoclast activity in vitro. In the present report the peripheral blood T major lymphocyte subsets were investigated in a group of genotyped type 1 Gaucher disease patients. A total of 31 patients were studied: 21 non-splenectomized (5 N370S homozygotes) and 10 splenectomized (of whom 1 was a N370S homozygote). The results show that non-splenectomized patients present a decrease in absolute numbers of peripheral blood T lymphocytes, specially the CD4+ T subset. However, when patients were analyzed with respect to the presence of bone disease, the number of CD8+ T lymphocytes was found to be statistically significantly lower in patients presenting bone involvement. Furthermore, lower numbers of CD8+ T lymphocytes were significantly correlated with higher levels of plasma tartrate resistant acid phosphatase (TRAP) activity, a putative marker of osteoclast cell activity. These in vivo findings are in agreement with the results reached in vitro by others. They provide an additional marker of disease severity in Gaucher disease. In the group of genotyped Gaucher disease patients, the majority of the N370S homozygous patients presented a clinically milder phenotype, including the absence of bone involvement, confirming earlier reports predicting that a number of these patients may remain undiagnosed. Collectively the homozygosity for the N370S mutation and normal T cell numbers may provide additional markers for the clinical heterogeneity of Gaucher disease. PMID- 10389596 TI - Cognitive approaches to delusions: a critical review of theories and evidence. AB - PURPOSE: To review critically the evidence for three contemporary theories of delusions. METHODS: The theoretical approaches to delusions proposed by Frith and colleagues ('theory of mind' deficits), Garety and colleagues (multi-factorial, but involving probabilistic reasoning biases) and Bentall and colleagues (attributional style and self-discrepancies) are summarised. The findings of empirical papers directly relevant to these proposals are critically reviewed. These papers were identified by computerised literature searches (for the years 1987-1997) and a hand search. RESULTS: The evidence does not unequivocally support any of the approaches as proposed. However, strong evidence is found to support modifications of Garety and colleagues' and Bentall and colleagues' theories. Studies have replicated a 'jumping to conclusions' data-gathering bias and an externalising attributional bias in people with delusions. There is preliminary evidence for a 'theory of mind' deficit, as proposed by Frith, although possibly related to a more general reasoning bias. Evidence for an underlying discrepancy between ideal and actual self-representations is weaker. CONCLUSIONS: A multi-factorial model of delusion formation and maintenance incorporating a data-gathering bias and attributional style, together with other factors (e.g. perceptual processing, meta-representation) is consistent with the current evidence. It is recommended that these findings be incorporated into cognitive therapy approaches. However, there are limitations to existing research. Future studies should incorporate longitudinal designs and first episode studies, and should not neglect the co-morbidity of delusions, including affective processes, or the multi-dimensional nature of delusions. PMID- 10389597 TI - Neuropsychological assessment of older adults with Down's syndrome: an epidemiological study using the Cambridge Cognitive Examination (CAMCOG). AB - OBJECTIVES: The Cambridge Cognitive Examination (CAMCOG) was designed for the general elderly population to assess cognitive impairments characteristic of dementia. CAMCOG yields a total score as well as separate scores on seven subscales. In this study the suitability of the CAMCOG for older adults with Down's Syndrome (DS) at the age of risk for dementia was assessed. DESIGN: A near total population sample of people with DS aged 30 years and over (range 30-65 years) living in a single Health Authority catchment area was identified and assessed using the CAMCOG. The use of an unselected sample ensured that the value of this assessment instrument could be established across all levels of disability in those with DS. A lower age limit of 30 years was used, as from this age significant Alzheimer-like neuropathology is present and prevalence rates of Alzheimer's disease begin to increase. METHODS: The CAMCOG was administered in a familiar setting together with other neuropsychological tests. Gender and age differences and the characteristics of those at the floor of the test were investigated. RESULTS: Of the 77 people with DS aged 30 years or older, 74 agreed to take part. Scores on the CAMCOG were well distributed, with only eight participants (11%) scoring 0 on the test. This contrasted favourably with performance on the Mini-Mental State Examination where there was a narrower range of scores and a higher percentage scoring 0. There was a significant difference in cognitive performance between younger (30-44 years) and older (45+ years) participants on the total CAMCOG score and on six out of the seven CAMCOG subscales. CONCLUSIONS: The CAMCOG, with minor modifications, is a useful test to assess those areas of cognitive function known to decline with dementia. Apart from those with pre-existing severe learning disability, severe sensory impairments and/or already advanced dementia, people with DS were able to score above the floor of the test. PMID- 10389598 TI - A cognitive distortion associated with eating disorders: thought-shape fusion. AB - OBJECTIVES: The primary objective of this study was to describe and investigate a cognitive distortion associated with eating psychopathology. This distortion, termed 'thought-shape fusion', is said to occur when merely thinking about eating a forbidden food increases the person's estimate of their shape or weight, elicits a perception of moral wrongdoing and makes the person feel fat. DESIGN: Two studies were conducted. The first was a psychometric study and the second utilized a within-participants experimental design. METHODS: In Study 1, thought shape fusion was assessed in a sample of 119 undergraduate students using a questionnaire. In Study 2, 30 students with high thought-shape fusion scores participated in an experiment designed to elicit the distortion. RESULTS: Thought shape fusion was found to be significantly associated with measures of eating disorder psychopathology. The questionnaire used to measure thought-shape fusion had high internal consistency, a good factor structure accounting for 46.2% of the variance and predictive validity. The results from Study 2 indicated that the distortion can be elicited under experimental conditions, produces negative emotional reactions and prompts the urge to engage in corrective behaviour (e.g. neutralizing/checking). This corrective behaviour promptly reduces the negative reactions. CONCLUSION: The results of the two studies indicate that the concept of thought-shape fusion is coherent, unifactorial and measurable. It is associated with eating disturbance and elicits negative emotional and behavioural responses. PMID- 10389599 TI - An investigation of attributional style in first-episode psychosis. AB - OBJECTIVE: The present study investigated covert and overt attributional styles in individuals experiencing a first episode of psychosis. It was hypothesized that those individuals experiencing paranoia, as operationalized by higher scores on the suspiciousness item of the Brief Psychiatric Rating Scale (BPRS) would perform differently on both covert and overt measures of attributional style when compared to those individuals who scored lower on the BPRS suspiciousness item. DESIGN AND METHODS: A cross-sectional design was used. The sample consisted of 62 participants (50 males and 12 females) from the Early Psychosis Prevention and Intervention Centre. The Pragmatic Inference Task (PIT) was used to measure covert attributional style, whereas the Attributional Style Questionnaire (parallel form; ASQpf) measured overt attributional style. The Rosenberg Self Esteem Questionnaire measured global self-esteem. Participants' positive, negative, and depressive symptoms were assessed by means of the BPRS, the Scale for the Assessment of Negative Symptoms, and the Beck Depression Inventory, respectively. RESULTS: Regression analyses found that less suspiciousness (p = .02) and more depression (p = .02) predicted higher internal attributions for negative events scores on the ASQpf. There was a trend (p = .07) for more suspicious individuals to endorse the self-serving bias (SSB) on the PIT, even despite the SSB not being large enough to be considered defensive. Verbal IQ emerged as a significant predictor of covert attributional style (p = .04). CONCLUSIONS: The findings suggest that increasing suspiciousness does predict attributional style in the early stages of psychosis, although the relationship appears weaker than in reports with more chronic psychotic patient samples. Longitudinal research is needed to ascertain whether attributional style is a stable characteristic in psychosis, or whether it fluctuates between periods of remission and active psychosis. PMID- 10389600 TI - Suppression and ritualistic behaviour in normal participants. AB - OBJECTIVE: Previous research has shown that normal and abnormal ritualistic behaviours do not differ in content. Rather, the differences between both categories of rituals pertain to characteristics such as frequency, intensity, discomfort and resistance. This study sought to investigate whether thought suppression is linked to these characteristics. DESIGN: Cross-sectional; questionnaires on thought suppression and rituals were administered to a sample of undergraduate students (N = 166). METHOD: Habitual suppressors (N = 20) and non-suppressors (N = 20), as measured by the White Bear Suppression Inventory, were selected and compared with regard to the characteristics of their rituals. RESULTS: Suppressors experienced their rituals as more intense, discomforting and resistance-provoking than did non-suppressors. There were no group differences in the content, frequency, and perceived senselessness of rituals. CONCLUSION: Although the cross-sectional nature of the present study precludes causal inferences, its findings are consistent with the view that chronic thought suppression may promote ritualistic behaviour. Clearly, the details of the link between thought suppression and rituals require further examination. PMID- 10389601 TI - Variceal haemorrhage and post-traumatic stress disorder. AB - OBJECTIVE: Post-traumatic Stress Disorder (PTSD) is thought to be relatively common following extremely distressing life-threatening events. Patients with liver cirrhosis can experience severe brisk variceal haemorrhage during which they vomit litres of blood and may exsanguinate. We predicted that a significant proportion of survivors would suffer from PTSD. DESIGN: PTSD assessment of 30 patients who had a haematemesis of more than four units of blood secondary to variceal bleeding and were fully conscious at the time of the bleed. METHOD: Structured Clinical Diagnostic Interview (SCID-DSM-III-R) and self-report measures. RESULTS: Most found the experience distressing, but only 1 out of 30 patients fulfilled DSM-III-R diagnostic criteria for PTSD. CONCLUSIONS: PTSD in a sample of patients who survived life-threatening variceal haemorrhage is much rarer than might reasonably have been anticipated. Possible reasons for this low prevalence of PTSD are discussed. PMID- 10389602 TI - Temporal stability of the Wisconsin Card Sorting Test in an untreated patient sample. AB - This study evaluates the temporal stability of the Wisconsin Card Sorting Test (WCST). The only previous similar study found unacceptably low test-retest stability in a non-patient elderly sample. In contrast, the present study utilizes 29 untreated obstructive sleep apnea patients who are more typical of persons likely to receive the WCST clinically. The 11 WCST scores examined demonstrated test-retest correlations from .34 to .83 (mean = .64). Six of 11 correlations were within .10 of the .80 criterion for clinical utility and there was minimal change across sessions. The current findings justify greater optimism regarding the test-retest reliability of the WCST. PMID- 10389603 TI - The economics of mental illness. PMID- 10389604 TI - Another nail in the coffin of confidentiality. PMID- 10389605 TI - Economic impacts of assertive community treatment: a review of the literature. AB - BACKGROUND: Assertive community treatment (ACT) is an extensively studied and widely imitated community support treatment model for severely mentally ill individuals. Several previous reviews have documented its favourable effects on clients and their families. This is the first review to focus on economic outcomes. METHODS: Nineteen randomized studies and 15 nonrandomized studies describing ACT programs were identified based on 2 criteria: 1) provision of services primarily in the community and 2) shared caseloads. Percentage reduction in hospital days was calculated for the 34 study sites where reported data allowed it. Multiple-regression methods were used to relate reduction in hospital days to program fidelity and other contextual factors. The impacts of ACT on emergency-room use, use of outpatient services, housing, costs, and other economic outcomes were also examined. RESULTS: Higher-fidelity programs appear to reduce hospital days by about 23 percentage points more than lower-fidelity programs (95% CI = -41.2, -5.2). The estimated regression coefficients imply that a high-fidelity program reduces hospitalizations by about 58% over 1 year if the alternative involves some type of case management and by 78% if it does not. ACT appears to increase the proportion of clients who live in independent housing situations, but the effect on use of supervised housing, and therefore on housing costs, is ambiguous. The effects on use of most other resources are inconsistent across studies. Overall, ACT appears to result in somewhat lower costs, whatever the perspective of analysis adopted. CONCLUSIONS: The most reliable cost offset to ACT treatment costs appears to be reduced hospital use. Using Quebec costs, an ACT program must enroll people with prior hospital use of about 50 days yearly, on average, to break even. As care systems evolve to reduce their reliance on hospitalization as a care modality with or without ACT, this threshold will become increasingly difficult to achieve. The primary justification for implementing ACT services will then become their clinical benefits. PMID- 10389606 TI - The valuation of productivity costs due to premature mortality: a comparison of the human-capital and friction-cost methods for schizophrenia. AB - OBJECTIVE: To compare productivity-cost estimates for schizophrenia-related premature mortality in Canada in 1996 using the human-capital (HC) approach and friction-cost (FC) method. METHODS: The number of deaths directly attributed to schizophrenia was combined with the estimated number of deaths attributable to schizophrenia from suicide. These premature deaths were valued using 2 methods: 1) the traditional HC approach, based on "potential" lost output to normal age of retirement, and 2) the FC method, based on finding a replacement worker. RESULTS: In 1996, there were 342 male and female preretirement deaths attributed to schizophrenia, directly or indirectly by suicide, in Canada. Most deaths were in males (78%) and by suicide (97%). The productivity cost of these deaths was estimated to be $105 million using the HC approach but only $1.53 million using the FC method. CONCLUSIONS: Productivity-cost estimates from the HC approach are substantially higher than those obtained from the FC method (69 times higher). In circumstances of unemployment, the HC approach is an overestimate of future productivity losses for premature mortality. PMID- 10389607 TI - The economic burden of schizophrenia in Canada. AB - OBJECTIVE: To estimate the financial burden of schizophrenia in Canada in 1996. METHOD: Using a prevalence-based approach, all direct health care costs, administrative costs of income assistance plans, and costs of incarceration attributable to schizophrenia were determined. Also included was the value of lost productivity associated with premature mortality and morbidity. In addition to using published papers and documents, direct contact was made with representatives from various provincial and federal programs for estimates of the direct health care and non-health care costs. RESULTS: The estimated number of persons with schizophrenia in Canada in 1996 was 221,000, with equal distribution between males and females. The direct health care and non-health care cost was estimated to be $1.12 billion in 1996. In addition, another $1.23 billion in lost productivity associated with morbidity and premature mortality was attributable to schizophrenia. CONCLUSIONS: The total financial burden of schizophrenia in Canada was estimated to be $2.35 billion in 1996. The largest category of cost was morbidity (52%), followed by acute care and psychiatric hospital admissions (14% and 10% respectively). Given the magnitude of these cost estimates, there are large potential cost savings with more effective management and control of this debilitating disease. PMID- 10389608 TI - Recruitment into psychiatry: increasing the pool of applicants. AB - OBJECTIVE: To demonstrate that it is possible to identify the cohort of students in their first year of medical school from which future psychiatrists will be recruited. METHOD: During a 3-year period, all first-year medical students at the University of Maryland completed a form indicating their specialty preference. Of those students, 403 pursued the regular psychiatry curriculum, and 34 participated in an enriched behavioural science and psychiatry program. Specialty was chosen after graduation. RESULTS: The higher the first-year student ranked psychiatry as a preferred specialty, the more likely the student was to choose psychiatry as a career after graduation. This was true both for students in the regular psychiatry program and for those in the enriched program. Students in the enriched program were significantly more likely to choose psychiatry as a career than were "regular" psychiatry students who gave psychiatry the same ranking in their first year. Freshman students who ranked psychiatry 4th or lower were not likely to choose psychiatry, no matter how much encouragement they received from their psychiatry departments. CONCLUSIONS: 1) Specialty preferences in the freshman year are predictive of future career choices. 2) An enriched medical school program in psychiatry can increase the number of graduates choosing careers in psychiatry. To help resource-poor medical schools increase the number of American medical graduates choosing psychiatry, the authors propose 2 inexpensive enriched programs. PMID- 10389609 TI - Clinical rating of compliance in chronic hemodialysis patients. AB - OBJECTIVE: To develop a clinical rating scale of treatment compliance for use in chronic hemodialysis patients and to test its reliability and validity. METHOD: Forty-eight of 65 patients undergoing hemodialysis treatment at the Ottawa General Hospital during June 1994 met criteria for inclusion and completed the study. Patients underwent a 10-15-minute interview, with 1 of 2 independent clinical interviewers, regarding diet, fluid intake, prescribed medication usage, smoking, alcohol or drug use, and hemodialysis treatment attendance. Following each interview, a predesigned 3-point rating scale evaluating compliance in each of 6 domains (yielding an 18-point total score) to the treatment regimen was completed. Compliance ratings on 10 patients assessed independently by both interviewers were used to establish scale reliability. Criterion validity was assessed by correlating compliance scale scores with 3 biological measures (weight gain [kg], K+ [mmol/l], and PO4 [mmol/l]). RESULTS: Reliability between clinical interviewers using the overall compliance scale score (Intraclass correlation coefficient = 0.825) as well as component subscales was high (kappa values, 0.33-1.00). Biological measures of compliance correlated well with each other but poorly with clinical ratings (range 0.01-0.16). Biological measures identified compliance as being poorer than that found with the clinical interview scale. CONCLUSIONS: The Compliance Rating Scale (CRS) was shown to be reliable but was not well-validated against selected biological measures. Discrepancies between these 2 methods of assessing compliance may be due to differing underlying compliance constructs or patient or interviewer biases. The CRS has value as a patient education tool in that it can be used to instruct patients regarding the benefits of adhering to the treatment regimen. PMID- 10389610 TI - Characteristics of problem gambling in a Canadian context: a preliminary study using a DSM-IV-based questionnaire. AB - OBJECTIVE: To develop a self-report instrument to assess diagnostic criteria and associated features of pathological gambling in order to learn more about the characteristics of individuals who seek treatment for gambling problems in a Canadian setting. METHOD: Fifty-seven adults seeking treatment for gambling problems at the Addictions Foundation of Manitoba were assessed. RESULTS: There was substantial variation in the endorsement of Diagnostic and Statistical Manual of Mental Disorders (DSM-IV) symptoms. Lying to family members or friends and "chasing" previous gambling losses were frequently reported, while more serious consequences (for example, relationship breakup, job losses) were less frequent. DSM-IV ratings were correlated (r = 0.59) with the South Oaks Gambling Screen. Many individuals reported gambling as a way to alleviate dysphoric mood, and 30% reported receiving mental health services in the past. Approximately 50% reported suicidal ideation, although recent suicide attempts were not common. CONCLUSION: These preliminary results of Canadian adults seeking treatment for gambling problems suggest a somewhat different profile from many United States studies, which often rely on older male pathological gamblers. More systematic investigation of the presence of major depression and other psychiatric disorders is warranted. Consistent with demographic data collected at the Addictions Foundation of Manitoba, it appears that video lottery terminals play a major role in the type of problem gambling experience seen in Canadian settings. PMID- 10389611 TI - Panic disorder in schizophrenia. AB - OBJECTIVE: To determine the frequency of panic attacks and panic disorder in patients with chronic schizophrenia or schizoaffective disorder. METHOD: Fifty three male outpatients meeting Diagnostic and Statistical Manual of Mental Disorders (DSM-IV) criteria for chronic schizophrenia or schizoaffective disorder were administered sections of the Structured Clinical Interview for DSM-IV (SCID). If panic attacks were reported, patients were queried about treatment and about onset relative to psychotic symptoms. RESULTS: Forty-nine patients were sufficiently organized to participate in the evaluation. Twenty-one (43%) experienced panic attacks, and 16 (33%) had current or past panic disorder. Eight (50%) of the 16 with panic disorder had been treated for panic. Substance dependence was not associated with having panic attacks or current or past panic disorder. Patients with paranoid schizophrenia were more likely than patients with schizoaffective or undifferentiated schizophrenia to have experienced panic attacks (57% versus 20%, chi 2 = 6.0, P < 0.02) or panic disorder (47% versus 10%, chi 2 = 6.9, P < 0.01). CONCLUSION: Panic attacks and panic disorder are common in men with schizophrenia or schizoaffective disorder. Panic disorder may be an overlooked comorbid diagnosis in patients with schizophrenia. PMID- 10389612 TI - Cognitive therapy for bipolar depression: a pilot study. AB - OBJECTIVE: While the efficacy of cognitive-behaviour therapy (CBT) for the treatment of acute unipolar major depression is well-documented, there is almost no data evaluating its utility in the treatment of bipolar depression. This pilot study compares the efficacy of CBT combined with mood-stabilizer pharmacotherapy for bipolar depression and CBT alone for unipolar depression. METHOD: A matched case control design was used to evaluate outcomes following 20 sessions of CBT in 11 depressed bipolar patients and 11 matched recurrent unipolar depressed control subjects. RESULTS: Bipolar depressed patients achieved similar levels of reduction in depressive symptoms following CBT, as did the unipolar depressed group. However, on measures of more pervasive dysfunctional attitudes, bipolar patients did not improve to the same degree. CONCLUSIONS: Preliminary findings suggest that CBT warrants further investigation as an effective psychosocial intervention for depression in bipolar patients already receiving ongoing mood stabilizing pharmacotherapy. PMID- 10389613 TI - Review board outcomes for involuntary patients in provincial psychiatric hospitals. AB - OBJECTIVE: To identify long-term trends and compare different psychiatric hospitals with regard to outcomes of involuntary certification. METHOD: Data on outcomes of involuntary certification were obtained from the London and St Thomas Psychiatric Hospitals for 1987 to 1997, from the Psychiatric Patient Advocate Office pertaining to 10 psychiatric hospitals for 1987 to 1993, and from published data from the North Bay Psychiatric Hospital for 1992 to 1994. Data were compared regarding outcome variables. RESULTS: There are variable rates and proportions among the psychiatric hospitals for outcomes of involuntary certification, including patients choosing to withdraw their applications to Review Boards, physicians cancelling involuntary certificates by completing Form 5s, and Review Boards rescinding certificates. Overall, few certificates were rescinded by the Boards, a trend that was even more pronounced in recent years. CONCLUSIONS: Patients who are in psychiatric hospitals on an involuntary basis are infrequently released from hospital as a result of a Review Board rescinding their certificate. PMID- 10389615 TI - Re: Gray and Keegan. PMID- 10389616 TI - Re: Mental health legislation and the right to appropriate treatment. PMID- 10389614 TI - Quetiapine: are we overreacting in our concern about cataracts (the beagle effect)? PMID- 10389617 TI - Restrictive measures in a psychiatric hospital on weekdays versus on weekends or holidays. PMID- 10389618 TI - Implications of lesion-clinical correlations in poststroke depression. PMID- 10389619 TI - Mania and donepezil. PMID- 10389620 TI - Tardive dystonia induced by risperidone. PMID- 10389621 TI - Undifferentiated carcinoma of the prostate with small cell features: immunohistochemical subtyping and reflections on histogenesis. AB - To investigate the histogenesis of undifferentiated carcinoma of the prostate with small cell features we analysed the expression of neuroendocrine (NE) markers, the androgen receptor (AR), and prostate-specific antigen (PSA) in 19 undifferentiated carcinomas of the prostate. The proliferative activity (MIB 1/Ki67) of the tumours was examined, and the clinical data reviewed. The results identified two groups: carcinomas in group 1 were positive for PSA and AR and negative for NE markers. The mean MIB-1 labelling index (LI) was 34.8% and the mean serum PSA value 56.4 ng/ml. Two of the 7 patients died within 12 months after tumour diagnosis. The tumours in group 2 were NE differentiated small cell carcinomas (SCC), which were negative for PSA and AR. The mean MIB-1 LI was 82.6% and the mean serum PSA value 7.1 ng/ml. Seven of the 10 patients died between 2 and 12 months after tumour diagnosis. Positive staining for NE markers in combination with negative staining for PSA and AR and a high MIB-1 LI substantiated the diagnosis of a NE-SCC. We suggest that this tumour has a stem cell origin and does not derive from a dedifferentiated adenocarcinoma or from benign NE cells of the prostatic epithelium. This clear distinction of NE-SCC from NE-negative undifferentiated carcinoma is in accordance with the differing biological behaviour and response to therapy of the two tumour entities. PMID- 10389622 TI - Myoepitheliomas of the skin and soft tissues. Report of 12 cases. AB - We describe 12 cutaneous and soft tissue myoepitheliomas, most of them in elderly patients. Morphologically the cutaneous and soft tissue myoepitheliomas revealed the same spectrum as their salivary gland counterparts. They were composed of a mixture of spindle, epithelioid and clear myoepithelial cells. Immunohistochemically they were positive to keratins and S-100 protein and reacted inconsistently with antibodies to smooth muscle actin. Morphologically they lacked any folliculo-sebaceous or apocrine differentiation. We believe that they are related to the eccrine type of cutaneous mixed tumours. Most cases had a benign behaviour, but 1 tumour metastasized, and the patient died of the tumour. Myoepitheliomas of soft tissues should be distinguished from other neoplasms with epithelial differentiation and from ossifying fibromyxoid tumour of soft parts, parachordoma and extraskeletal myxoid chondrosarcoma. PMID- 10389623 TI - Immunohistochemical study of cytochrome P450 2C and 3A in human non-neoplastic and neoplastic tissues. AB - Organ and cellular distribution and expression constancy of microsomal cytochrome P450 (CYP) 2C and 3A in humans were studied with new polyclonal antibodies to CYP2C (MP-1) and 3A (NF-2) active in formalin-fixed, paraffin-embedded tissues. Antibodies were raised against purified human CYP2C9 and CYP3A4. On western blotting, MP-1 reacted with 2C8, 2C9, 2C18 and 2C19, and NF-2 with 3A4. In both frozen and paraffin sections, hepatocytes showed diffuse immunoreactivity with MP 1 and centrilobular staining with NF-2. In-paraffin sections of 40 kinds of nonneoplastic tissues, epithelium of the small and large intestine, bile duct, nasal mucosa, kidney and adrenal cortex stained positively with both MP-1 and NF 2 antibodies. Epithelium of gastric fundic glands, salivary glands, tracheobronchial glands, Brunner's glands, the prostate, uterine cervix and nasopharynx showed definite reactivity with MP-1. Epithelium of the gastric mucosa with intestinal metaplasia, duodenum, gallbladder and intercalated ducts of the pancreas and chief cells of the parathyroid and the corpus luteum of the ovary reacted with NF-2. Among the neoplastic tissues, MP-1 reacted with pleomorphic adenoma of the salivary gland and carcinomas of six different organs, and NF-2 with those of 7 different organs. These results indicate that CYP2C and CYP3A are distributed widely and organ specifically, as well as being variably expressed in neoplastic and normal states. PMID- 10389624 TI - Induction of smooth muscle cells in the fibrous capsule of human hepatocellular carcinoma but not in the septa of hepatic cirrhosis. AB - We examined the expression of smooth muscle cytoskeleton in spindle-shaped cells in the capsule of hepatocellular carcinoma (HCC) and the septa of liver cirrhosis (LC). Serial sections of livers resected from 11 patients were stained with monoclonal antibodies against vimentin, desmin, smooth muscle actin (1A4, HHF35, CGA7) and smooth muscle myosin heavy chain isoforms (SM1, SM2). Capsular spindle shaped cells exhibited a cytoskeletal feature indicative of intermediately differentiated smooth muscle cells. Computer-assisted morphometry revealed that the proportions of 1A4-, HHF35-, CGA7- and SM1- positive areas to vimentin positive area were 88.0+/-11.0%, 50.8+/-17.4%, 25.3+/-16.4% and 19.4+/-12.4% (n=11) in main tumours and 86.6+/-9.4%, 50.9+/-18.7%, 21.1+/-12.3% and 17.6+/ 9.7% (n=12) in daughter tumours, indicating that spindle-shaped cells are heterogeneous in cytoskeletal expression. Septal spindle-shaped cells in LC lacked the cytoskeletal proteins specific to differentiated smooth muscle cells (CGA7, SM1, SM2 and desmin). Electron microscopically, capsular spindle-shaped cells contained more microfilaments and less rough endoplasmic reticulum than do septal cells. Intermediately differentiated smooth muscle cells are induced in the capsule of HCC but not in the septa of LC, suggesting a role for stromal interaction by tumour cells in the induction of smooth muscle cells. PMID- 10389625 TI - Strongly reduced expression of the cell cycle inhibitor p27 in endometrial neoplasia. AB - In the present study we investigated the expression of the cell cycle inhibitor p27 in endometrial neoplasia using immunohistochemistry with a p27-specific antibody. Expression of p27 in endometrial carcinomas was compared with expression in the normal endometrium throughout the cycle. Normal endometrial cells showed strong nuclear expression of p27. Expression was present throughout the cycle and was stronger during the secretory phase. We found strongly reduced or abolished expression of p27 in endometrial carcinoma (85.3% of cases). The 41 tumours analysed were classified according to p27 staining intensity and percentage of positive cells into the following categories of p27 expression: negative/very low (56.0%); low (29.3%); moderate (14.7%) and high (0.0%). All the p27-positive tumours were well-differentiated endometrioid carcinomas of malignancy grade G1. Comparison with the p53 status showed that all tumours with strong p53 expression had low/negative p27 staining, while those that were positive for p27 had negative/low p53 staining. Reduced or absent p27 levels were also observed by Western blot analysis both in tumour samples and in HEC-1B endometrial adenocarcinoma cells. It thus seems that p27 expression is essential for the control of normal endometrial proliferation, and reduced or absent p27 expression may be an important step in endometrial carcinogenesis. PMID- 10389626 TI - Expression of melanocyte-associated markers gp-100 and Melan-A/MART-1 in angiomyolipomas. An immunohistochemical and rt-PCR analysis. AB - Angiomyolipomas are tumours of uncertain histogenesis, most often occurring in association with the kidney. A characteristic finding is their reactivity with HMB-45, a monoclonal antibody to the melanocyte-associated antigen gp-100. We tested 18 angiomyolipomas for their reactivity with A103, a monoclonal antibody to Melan-A (MART-1), another melanocyte-associated marker, and compared it with HMB-45. All cases were positive with both antibodies, yet most cases showed a more homogeneous staining pattern with A103. Normal kidney was immunohistochemically negative for both antibodies. We also performed RT-PCR assays for gp-100 and Melan-A in 4 of the 18 angiomyolipoma samples and in three normal kidney samples. All 4 angiomyolipoma specimens revealed mRNA for both melanocyte differentiation markers. gp-100 mRNA was found in the samples of normal kidney, but Melan-A mRNA was not. Our study shows that angiomyolipomas express the melanocyte-associated antigens Melan-A and gp-100 at the protein and at the mRNA level, suggesting a true expression of these antigens rather than cross-reacting epitopes. Based on the mRNA expression pattern, immunohistochemical analysis is the preferred method for the detection of gp-100, while Melan-A can be used at the protein and mRNA levels. Our study demonstrates that A103 is a useful marker for the diagnosis of angiomyolipomas. PMID- 10389627 TI - Analysis of subclonal expansion of colorectal carcinomas by flow cytometry. AB - DNA heterogeneity of colorectal carcinomas has been investigated by flow cytometry, most studies have focused on the clinical usefulness of DNA ploidy analysis. Since cancers consist of predominant subclones with proliferative advantage due to clonal expansion, we attempted to analyse the clonal expansion of colorectal carcinomas within a tumour by measuring DNA ploidy. The DNA ploidy and heterogeneity of multiple fresh samples obtained from 164 colorectal adenocarcinomas were analysed by flow cytometry. Each tumour was divided into an average of six specimens, which were analysed separately. For 146 of the tumours (89%) at least one DNA aneuploid population was found within the cancer tissue examined. DNA multiploidy was detected in 26 cases (17.8%) among the cancers with aneuploidy. Based on the DNA index (DI), hypertriploid aneuploidy (1.7 2 mg/l (resistant). The following strains were tested: 61 Escherichia coli, 12 Klebsiella pneumoniae, 7 Proteus mirabilis, 21 Serratia marcescens, 4 Enterobacter cloacae, 21 Pseudomonas aeruginosa, 21 Staphylococcus. aureus (resistant to methicillin) and 15 Enterococcus spp. Clinafloxacin, ciprofloxacin, ofloxacin and norfloxacin activities were evaluated by agar dilution using Mueller-Hinton agar according to NCCLS recommendations. Of the 162 isolates, 16 (9.8%) were intermediate and 146 (90.1%) resistant to ciprofloxacin. 95 of the 162 strains (58.6%) were susceptible, 27 (16.7%) intermediately susceptible, and 40 strains (24.7%) were resistant to clinafloxacin. The percentage susceptible to clinafloxacin was 65.6% for E. coli, 75% for K. pneumoniae, 71.4% for P. mirabilis, 28.6% for S. marcescens, 75% for E. cloacae, 33.3% for P. aeruginosa, 90.5% for S. aureus and 40% for Enterococcus spp. Clinafloxacin was active against 58.6% of the ciprofloxacin-resistant clinical isolates tested. It was particularly active against S. aureus strains resistant to both ciprofloxacin and methicillin. PMID- 10389647 TI - In vitro evaluation of intracellular activity of antibiotics against non-typhoid Salmonella. AB - Non-typhoid salmonellae are the most common causative organisms of bacterial enteritis in children. Clinical studies have failed to show any influence of various antibiotics on the natural course of acute salmonella enteritis. Poor penetration of antibiotics into phagocytic cells that contain intracellular Salmonella spp., and possible intracellular antibiotic inactivation have been considered as possible reasons for this. In this study, we used an in vitro model to assess the intracellular activity of antibiotics against non-typhoid salmonellae. The survival of intracellular Salmonella spp. in P388D1 cells, a mouse macrophage cell line was measured in the presence of various antibiotics. Except for gentamicin, which entered phagocytes poorly, ofloxacin, azithromycin, chloramphenicol and three beta-lactam antibiotics, ampicillin, cefixime and ceftriaxone, exhibited bacteriostatic activity against susceptible intracellular Salmonella spp. at an extracellular concentration equal to the minimal inhibitory concentration (MIC). At a concentration of 10 x MIC, neither chloramphenicol nor the three beta-lactam antibiotics produced a bactericidal response; however, both ofloxacin and azithromycin were bactericidal after 8-24 h of incubation. The results showed that fluoroquinolones and new macrolides were more efficient than the other antibiotics in eradicating intracellular salmonella and might be useful agents for the treatment of non-typhoid salmonella enteritis in children. Clinical trials should be considered. PMID- 10389649 TI - Commentary on the MAFF technical report: a review of antimicrobial resistance in the food chain. Ministry of Agriculture Fisheries and Food. PMID- 10389648 TI - Inhibition of the in vitro growth of Plasmodium falciparum by acyclic nucleoside phosphonates. AB - Forty-eight acyclic nucleoside phosphonates (putative prodrugs of acyclic nucleoside triphosphate inhibitors of DNA replication) have been evaluated for in vitro antiplasmodial activity. Only certain purine derivatives with a hydroxyl group attached to the acyclic sugar moiety displayed antiplasmodial activity. The two most active analogs were (S)-9-(3-hydroxy-2-phosphonylmethoxypropyl)adenine ((S)-HPMPA, IC50=0.18+/-0.07 microM) and (S)-3-deaza-HPMPA (IC50=0.29+/-0.08 microM). Their cyclic derivatives, containing an ester bond between the phosphonate and the hydroxyl group, were slightly less active. All tested compounds that lacked the hydroxyl group, including potent antiretrovirus analogs such as 9-(2-phosphonylmethoxyethyl)adenine (PMEA) and the (S)-HPMPA derivatives (R)-PMPA and (S)-FPMPA, did not show any activity, even at very high concentrations ( >250 microM). Similarly, pyrimidine analogs of (S)-HPMPA, such as (S)-HPMPT, (S)-HPMPU and the anti-herpesvirus analog (S)-1-(3-hydroxy-2 phosphonylmethoxypropyl) cytosine ((S)-HPMPC), were devoid of any antiplasmodial activity. In addition, 11 acyclic nucleoside (non-phosphorylated) analogs--which in contrast to the acyclic nucleoside phosphonates require the presence of a monophosphorylating enzyme for the first activation step--were tested. None of them inhibited the growth of the parasite. In short three chemical entities seem to be imperative for antiplasmodial activity: a purine base, a hydroxyl group in the acyclic side chain and a phosphonate group terminating this chain. PMID- 10389650 TI - In vitro activity of several antimicrobial agents against 1003 isolates of Streptococcus pyogenes collected from Western Canada. AB - Streptococcus pyogenes is a common pathogen which may be associated with significant morbidity and mortality. Recent information is not readily available, in Canada, regarding the susceptibility of clinical isolates to penicillin, extended spectrum and/or newer agents. We collected and tested 1003 isolates of S. pyogenes to seven antimicrobial agents and found the following susceptibility rates: azithromycin 97%, ceftriaxone 100%, ciprofloxacin 99.4%, clarithromycin 98.5%, clindamycin 99.9%, erythromycin 96.5% and penicillin 100%. These results indicate that antimicrobial resistance is not yet a problem of S. pyogenes in Canada. PMID- 10389651 TI - Resistance problems in two university hospitals in Denmark. AB - In 1997, 80 and 89% of Staphylococcus aureus isolated from university hospitals in Odense and Hvidovre respectively were resistant to penicillin and 32.0 and 41% of Escherichia coli were resistant to ampicillin. There were low incidences of methicillin resistance in S. aureus (<1%), penicillin resistance in Streptococcus pneumoniae (3%),and gentamicin in E. coli (2%). These figures might reflect the low use of antibiotics in Denmark. PMID- 10389652 TI - Summary of the first international symposium on viral hepatitis. PMID- 10389653 TI - Utilization of transgenic mice replicating high levels of hepatitis B virus for antiviral evaluation of lamivudine. AB - A recently developed transgenic mouse strain which expresses high levels of hepatitis B virus (HBV) was studied as a model for evaluation of potential chemotherapeutic agents. Lamivudine ([-]2'-deoxy-3'-thiacytidine), known to reduce hepatitis B viremia in human patients, and zidovudine (3'-azido-3' deoxythymidine), previously shown to be ineffective for HBV infections in man, were used in parallel in this transgenic animal model. Orally administered lamivudine at dosages of 100, 50, and 25 mg/kg per day given once a day for 21 days significantly decreased serum and liver HBV DNA titers in a dose-responsive manner. Zidovudine (approximately 22 mg/kg per day) administered in the drinking water for 21 days was not effective in reducing these HBV parameters as compared to transgenic placebo-treated controls. The serum HBV DNA titers rebounded to high levels 1 week after cessation of lamivudine treatment. Male and female mice responded in a similar manner to these therapies. The results using this transgenic mouse model were similar to what would be predicted from treatment of HBV-infected human patients with lamivudine and zidovudine, and indicate these mice may be useful as a small animal chemotherapeutic model for study of potential HBV inhibitors. PMID- 10389655 TI - Synthesis and anti-influenza virus activity of novel pyrimidine derivatives. AB - Efficient synthetic routes of 2-amino-4-(omega-hydroxyalkylamino)pyrimidine derivatives were investigated in relation to the anti-influenza virus activity of these compounds. The derivatives in which cyclobutyl and cyclopentyl groups were introduced to the beta-position of the aminoalkyl group (especially the cyclobutyl group substituted by a phenylalkyl group at the 3'-position) resulted in improved antiviral potency: i.e. an average 50% effective concentration for inhibition of plaque formation (EC50, microM) of 0.1-0.01 microM for both types A and B influenza virus. The antiviral efficacies were in the order of amino group > hydroxyiminomethyl group > halogen substitution at the 5-position, and chlorine or methoxy group > hydrogen at the 6-position of the pyrimidine ring. The antiviral indices of these compounds were 2-6 with respect to the 50% inhibitory concentration for cell proliferation (IC50, microM) for growing cells, but > 500 to > 10(4) with respect to the IC50 for stationary cells, indicating that these compounds may be efficacious for the topical treatment of influenza virus infection. PMID- 10389654 TI - Induction of apoptotic cell death by IFNbeta on HPV-16 transformed human keratinocytes. AB - Apoptosis, or 'programmed cell death' is a process of general biological relevance with implications in several physiological and pathological conditions of the skin. However, little is known about its induction in keratinocytes by regulator agents. In this work we demonstrate that IFNbeta, but not IFNalpha, selectively induces programmed cell death in HPK-Ia cells, a line derived from human keratinocytes transformed with HPV-16 DNA. This IFNbeta-triggered apoptosis is strictly dependent on a serum-induced partially differentiated phenotype; it occurs through the activation of a check point in the early 'S' phase, where the cells are arrested and eventually driven to apoptosis. These data indicate that apoptosis may be induced in keratinocytes by a regulator agent combined with a differentiating stimulus. PMID- 10389656 TI - Evaluation of anti-herpesvirus activity of (1'S,2'R)-9-[[1',2' bis(hydroxymethyl)cycloprop-1'-yl]methyl]- guanine (A-5021) in mice. AB - The anti-herpesvirus activity of (1'S,2'R)-9-[[1',2'-bis(hydroxymethyl)cycloprop 1'-yl]methyl]guani ne (A-5021) was evaluated in murine cells and in several murine models of herpes simplex virus (HSV) infection. Against HSV type 1 (HSV 1), A-5021 was 15-30- and 30-60-fold more active, and against HSV type 2 (HSV-2), it was 2- and 8-fold more active than acyclovir and penciclovir in Balb/3T3 cells, respectively. When antiviral compounds were administered orally (once daily) to mice infected intraperitoneally with HSV-1 (Tomioka), A-5021 was more active than acyclovir or famciclovir in spite of its relatively low oral bioavailability. A-5021 was as active as penciclovir when the antiviral compounds were given intravenously (three times daily) to mice infected intraperitoneally with HSV-2 (186). In mice with a cutaneous HSV-1 (KOS) infection, three times daily oral therapy with A-5021 at 25 mg/kg per day produced more significant reduction in severity of skin lesions than equivalent treatment with acyclovir or famciclovir. In mice infected intracerebrally with HSV-1 (Tomioka), complete survival was observed in the group treated intravenously with A-5021 at 25 mg/kg per day (three times daily), while more than 50% of mice died in the groups treated intravenously with acyclovir of up to 100 mg/kg per day (three times daily). Moreover, A-5021 was more effective than acyclovir in clearing infectious virus from the brain. These findings demonstrate that A-5021 has potent anti-HSV activity in several murine models. PMID- 10389657 TI - Automated sleep staging systems in rats. AB - One of the major inconveniences encountered in sleep studies is the time consuming labor involved in equating visual analysis of physiological recordings (EEG, EMG, EOG, ...) to an appropriate state of vigilance. The explosion of computer technology is responsible for the emergence of several automated sleep wake staging systems to supplement human analysis. Conversely to human sleep analysis, rat sleep is characterized by the absence of consensus about numerous elements constituting the sleep-wake staging systems used to build a hypnogram (recording position, length of epoch, number and definition of the vigilance state discriminated, ...). If justified, the choices of the parameters involved by each system generally result from various viewpoints (physiology, mathematics, electronics, ...). The diversity generated by the liberty offered the investigator in building a system excludes any rigorous comparison between systems. Nevertheless, this variety can also be viewed as a representative of the effervescence of research in the field of sleep, and as a catalyst for new ideas. PMID- 10389659 TI - An algometer for intraoral pain tolerance measurements. AB - An algometer was developed to provide a variable probing force (0-200 g force) which could be used intraorally. This algometer consisted of an autoclavable probe handpiece attached to an optical encoder, which recorded probing force to a computer when a button was pressed. The probe handpiece included a 0.40 mm diameter hemispherical tip which was placed in contact with the experimental site. The probe tip was pressed against the tissue with increasing force until the subject pressed the button, at which point the pain tolerance (PT) value was recorded by the computer. Intraoral soft tissue, PT values were obtained from nine healthy adult subjects during 6 weekly visits to determine the reproducibility of PT measurements. Five gingival sulcus sites and three gingival surface sites, all adjacent to the maxillary premolars constituted the experimental area. The reproducibility of PT values using the force stimulus from the algometer was evaluated using interclass correlation coefficients (R) for each of the eight sites. Visits 1 and 2 were training and calibration visits. Visits 3 through 6 were experimental visits. The R values ranged from 0.40 to 0.79 when data from all six visits were included. R values for Visits 3 through 6 were 0.63-0.97 indicating good to excellent correlation after subjects became familiar with the procedure. A complete repeated measures analysis of variance (ANOVA) showed no significant interaction between site and visit. Duncan's multiple range test was used to compare sensitivity across the eight sites. The results indicated that the three most anterior sites were significantly (P < 0.05) more sensitive than four of the posterior sites. When the sites were grouped into: (1) gingival surface sites; (2) mid-facial sulcus sites; and (3) interproximal sulcus sites, no significant differences were found in their PT values. The algometer is well suited for intraoral investigations because of its precision, computerized data entry and easily positioned, autoclavable handpiece. This new algometer may be useful for sensory and pain studies for other parts of the body. PMID- 10389658 TI - Application of positron emission tomography to determine cerebral glucose utilization in conscious infant monkeys. AB - Cerebral glucose metabolism has been used as a marker of cerebral maturation and neuroplasticity. In studies addressing these issues in young non-human primates, investigators have used positron emission tomography (PET) and [18F]2-fluoro-2 deoxy-D-glucose (FDG) to calculate local cerebral metabolic rates of glucose (1CMRG1c). Unfortunately, these values were influenced by anesthesia. In order to avoid this confounding factor, we have established a method that permits reliable measurements in young conscious vervet monkeys using FDG-PET. Immature animals remained in a conscious, resting state during the initial 42 min of FDG uptake as they were allowed to cling to their anesthetized mothers. After FDG uptake, animals were anesthetized and placed in the PET scanner with data acquisition beginning at 60 min post-FDG injection. FDG image sets consisted of 30 planes separated by 1.69 mm, parameters sufficient to image the entire monkey brain. Our method of region-of-interest (ROI) analysis was assessed within and between raters and demonstrated high reliability (P < 0.001). To illustrate that our method was sensitive to developmental changes in cerebral glucose metabolism, quantitative studies of young conscious monkeys revealed that infant monkeys 6-8 months of age exhibited significantly higher 1CMRG1c values (P < 0.05) in all regions examined, except sensorimotor cortex and thalamus, compared to monkeys younger than 4 months of age. This method provided high resolution images and 1CMRG1c values that were reliable within age group. These results support the application of FDG-PET to investigate questions related to cerebral glucose metabolism in young conscious non-human primates. PMID- 10389660 TI - Caloric stimulation of ampullar receptors: a new method to produce mechanically evoked responses in frog semicircular canals. AB - A microthermistor positioned close to the exposed posterior semicircular canal in isolated labyrinth preparations of the frog was used to stimulate the sensory organ. Our results indicated that, depending on the position of the heater, the induced endolymphatic convection currents may result in either excitatory or inhibitory cupular deflections and thus in a modulation of ampullar receptor resting activity. Other possible thermal-dependent mechanisms, such as a direct action of the stimulus on vestibular sensors or endolymphatic volume changes, had, in the present experimental conditions, a minor role. Caloric stimulation could therefore represent a novel method to stimulate the semicircular canals 'in situ'. PMID- 10389661 TI - Demonstration of two distributions of vesicle radius in the dopamine neuron of Planorbis corneus from electrochemical data. AB - An electrochemical model to calculate the relative size and neurotransmitter concentration of individual nerve cell vesicles is presented to examine potentially different types of vesicles in Planorbis corneus. Amperometric current transients resulting from individual exocytosis events detected from single cells contain the information necessary to quantify vesicular neurotransmitter amount and to estimate other important cellular properties such as vesicular neurotransmitter concentration and vesicle radius. Use of a simplifying assumption that the cross-sectional area of the contents of each release event is the apparent electroactive area of the electrode and that the shape of the decreasing phase of each current transient follows Cottrell-like behavior, the Cottrell equation and Faraday's law can be combined to yield expressions for relative vesicle radius and neurotransmitter concentration. This analysis has been applied to data obtained from the cell body of the giant dopamine neuron of the pond snail P. corneus. The histogram of vesicular dopamine concentration reveals a single wide distribution and the histogram of vesicle radius reveals a bimodal radius distribution. These data strongly suggest two distinct classes of vesicle radius in the P. corneus neuron lead to the bimodal distribution of amount released reported earlier. PMID- 10389662 TI - Calculation of transmitter concentration in individual PC12 cell vesicles with electrochemical data and a distribution of vesicle size obtained by electron microscopy. AB - A mathematical model is described to accurately calculate vesicle size and neurotransmitter concentration distributions from electrochemical data. This model uses parameters from electrochemical exocytosis data obtained from PC12 cells in culture to calculate a size distribution that is then correlated to the size of vesicles obtained by electron microscopy. The relative standard deviation of the size distribution calculated from electrochemical data is 25% which matches the relative standard deviation of the vesicle size distribution measured by electron microscopy. The distribution calculated from electrochemical data is normalized to the vesicle size distribution of PC12 cell vesicles obtained from electron microscopy. Calculation of a vesicular catecholamine concentration histogram from the normalized size data and electrochemical parameters is then possible for individual exocytosis events. The average vesicular catecholamine concentration for PC12 cells as calculated by this method is 148+/-7 mM. More importantly, there is a distribution of concentration rather than a constant value. Additionally, the model permits evaluation of the concentration of transmitter in each individual vesicle and vesicle size for each vesicle from electrochemical data when the overall vesicle size distribution is known. PMID- 10389663 TI - Morphometric analysis of human mastication. AB - Motor activity studies usually involve analyzing behavior characteristics that are easily observed and scored. The advantage of these methods is that behavioral variation can be rapidly assayed. The disadvantage is that considerable behavioral information is lost. At the other extreme are descriptive kinematic studies that provide detailed quantification of behavior characteristics. The disadvantage of these descriptive studies has been the time-intensive nature of data acquisition and analysis. Recent technological advances provide means for scoring and analyzing many motor activity characteristics simultaneously without a significant increase in scoring and analysis time. The methods described in this paper provide an expeditious means of preserving the richness of motor activity for experimental and clinical neurobehavioral research purposes. PMID- 10389664 TI - A method to quantify glial fibrillary acidic protein immunoreactivity on the suprachiasmatic nucleus. AB - The aim of the present study was to develop a quantitative method to measure the immunoreactivity of the glial fibrillary acidic protein on the suprachiasmatic nucleus of hamster. For this purpose, optical microscopy images from brain sections processed for glial fibrillary acidic protein immunostaining were digitised under different light conditions. Image treatment and immunoreactivity quantification were performed following five different methods using the program Adobe Photoshop. The results were analysed in order to determine the ability of each method to differentiate immunoreactivity levels, and their susceptibility to the light conditions during image acquisition. Four of the five methods were found to be susceptible to the tested light conditions, while the other was not. This last method, which permits detection of differences in immunoreactivity between the different sections, was considered for further quantification of glial fibrillary acidic protein immunoreactivity on the suprachiasmatic nucleus. PMID- 10389665 TI - Volumetric MRI measurements of the tree shrew hippocampus. AB - Protocols suitable for repeated magnetic resonance imaging (MRI) studies of the tree shrew's brain were established. This included the development of (i) a technique for prolonged inhalation anesthesia by endotracheal intubation; (ii) a reproducible fixation of the animal's head in a stereotaxic frame and finally (iii) the set-up of the hardware (rf coil) and software (MRI sequences) of the MRI system. The endotracheal intubation as well as the repeated and prolonged anesthesia showed no complications. The in vivo measurements of the tree shrew's hippocampal formation revealed a high reproducibility. Right and left hippocampal volume was determined as 85.2 mm3 +/- 8% and 87.4 mm3 +/- 10%, respectively. The utility of MRI in delineating alterations in brain anatomy was demonstrated in three animals receiving cortisol via the drinking water (5 mg/animal/day). After a 4-week treatment, in two of the three tree shrews a reduction in hippocampal volume was observed. Thus, the MRI protocols used here allow for repeated and non invasive measurements of changes in hippocampal anatomy within the same animal and to monitor the temporal dynamics of structural alterations within this brain structure. PMID- 10389666 TI - An automatic 3D tracking system with a PC and a single TV camera. AB - We developed a novel system, for tracking a freely moving object (animal) in a limited 3D space, based on a closed loop TV and PC. Instead of using a pair of TV cameras to capture the 3D scene through the pair of its orthogonal projections, the system captures these projections by using a single TV camera. The direct view of the scene (e.g. its ground plan, i.e. projection onto the x-y plane) is captured together with its side projection, (i.e. projection onto the z-x plane) observed in a slanted mirror. The system uses a simple detection algorithm and hardware that were originally developed for 2D tracking of a single, contrast object on a stable and homogeneous background. All three coordinates of the tracked object are evaluated, displayed and stored in real-time, at 25 interlaced frames (samples) per second. The system was implemented on an IBM PC enhanced by an universal I/O board (Kaminsky, Yu, Krekule, I. Universal multifunctional IBM PC I/O board for clinical examinations and experimental research in neuroscience. Physiol Res 1994;43:193-199) and tested by tracking a LED marker, a small living fish in a water tank and a pair of LED markers which were activated one at the time in alternating TV frames thereby demonstrating the ability to simultaneously track two or more objects in 3D. PMID- 10389667 TI - An unsteady platform test for measuring static equilibrium in mice. AB - An unsteady platform test is presented in which mice must remain still on a narrow surface in order to prevent a fall. The mouse spontaneous mutation, Lurcher, causing cerebellar cortical degeneration, was evaluated on the unsteady platform, requiring balance in a stable body position (static equilibrium), as opposed to the stationary beam test, in which the animals are free to move on a larger surface (dynamic equilibrium). Lurcher mutants spent less time and had a higher number of slips than controls on the unsteady platform. In contrast, Lurcher mutants did not differ from controls for latencies before falling and distance travelled on the stationary beam. These results are discussed in terms of the possible involvement of two cerebellar circuits in motor control. PMID- 10389668 TI - Why neuroradiologists should consider very-high-field magnets for clinical applications of functional magnetic resonance imaging. PMID- 10389669 TI - Cognitive functional magnetic resonance imaging at very-high-field: eye movement control. AB - The oculomotor system, which optimizes visual interaction with the environment, provides a valuable model system for probing the building blocks of higher-order cognition. Attention shifting, working memory, and inhibition of prepotent responses can be investigated in healthy individuals and patients with brain disorders. Although the neurophysiology of the oculomotor system has been well characterized at the single-cell level in nonhuman primates, its functional architecture in humans determined by evoked response procedures and studies of patients with focal lesions has been limited. Available evidence points to a widely distributed set of neocortical and subcortical brain regions involved in the control of eye movements, including brain stem, cerebellum, thalamus, striatum, and parietal and frontal cortices. The advent of functional magnetic resonance imaging provides a noninvasive manner of localizing, at high spatial resolution, the brain systems that subserve different aspects of sensory and cognitive processes in humans. Functional magnetic resonance imaging studies have already delineated the brain systems subserving sensorimotor and cognitive control of eye movements in adult and pediatric populations. Hence, the combination of functional magnetic resonance imaging and eye movement procedures can be used to probe the integrity of the brain in neurological and psychiatric disorders as well as provide a window into the changes in brain function subserving cognitive development. PMID- 10389670 TI - Modeling the mind: very-high-field functional magnetic resonance imaging activation during cognition. AB - This article describes how fMRI can be used to examine the large-scale networks of cortical areas that subserve high-level cognition, such as sentence comprehension and visual thinking. The findings from a number of studies show that the qualitative and quantitative nature of the cognitive processes determines which cortical areas are activated (the network constituency) and the degree to which each network member is activated. For example, during sentence comprehension, activation in the left posterior temporal region and the inferior frontal gyrus, as well as their right hemisphere homologs, increases as a function of the linguistic complexity of the sentence. Such findings indicate that cognition emerges from the collaboration among the multiple cortical areas that compose the large-scale networks, rather than from the aggregate of autonomously functioning modules. The patterns of activation also show systematic shifts in the activity of a network during the spontaneous recovery of function by stroke patients, demonstrating cortical plasticity in adults. Finally, the article describes some simulation models that relate the information processing activity of a computational system to its resource consumption. This construct enables a mapping from the functional properties of the cognitive systems to the biological substrate that is reflected in fMRI. PMID- 10389671 TI - Clinical rationale for very-high-field (3.0 Tesla) functional magnetic resonance imaging. AB - The recent development of integrating very-high-field magnets (3.0 T) into clinical scanners was driven by the demonstration that both functional and anatomic information can be derived reliably at high spatial and temporal resolution. As very-high-field magnetic resonance imaging now approaches its entry into the clinical arena with a product by one major scanner manufacturer that is to be considered for Food and Drug Administration clearance, it is timely to consider the clinical applications that are likely to make such new technology significant in medical imaging. The rationale for very high field is based on the mechanism of blood oxygenation level-dependent contrast and the need for reliable functional studies on individual patients. The use of functional magnetic resonance imaging in the setting of presurgical planning is demonstrated in a number of different clinical scenarios. Such cases covering both pediatric and adult patients indicate that 3.0-T functional magnetic resonance imaging has an important role in neuroradiology. PMID- 10389672 TI - High temporal resolution functional magnetic resonance imaging at very-high field. AB - Functional magnetic resonance imaging (fMRI) is a novel neuroimaging technique that has enjoyed explosive growth during the past 7 years. It can be implemented relatively easily on many already existing MRI systems, it is noninvasive, and functional images may be obtained within tens of seconds. However, it measures a secondary effect of neuronal activity, the blood oxygen concentration (the blood oxygen level-dependent, or BOLD, effect), which is a somewhat sluggish and blurred measure of the actual time course of neuronal activity. Here we discuss the present limitations to temporal resolution and the degree to which they can be overcome by using specific assumptions about the coupling between neuronal activity, blood flow, and oxygen metabolism, or by specific experimental designs, in particular time-resolved fMRI. PMID- 10389673 TI - High spatial resolution functional magnetic resonance imaging at very-high magnetic field. AB - Although neuroimaging methods have been used successfully to map large-scale neurocognitive networks distributed across the human cortex, functional mapping and differentiation of localized brain organization within a small structure has been limited by inadequate sensitivity for high spatial resolution imaging. Functional magnetic resonance imaging (fMRI) technique based on blood oxygenation level-dependent (BOLD) contrast has become one of the most useful neuroimaging techniques. It has been used extensively to study human brain function from sensory perception to cognitive performance. However, the majority of these studies used a relatively low spatial resolution (typically with a voxel size of 3.1 x 3.1 x 5.0 mm3), which is incapable of mapping on the millimeter and submillimeter spatial scale. In this article, we review the technical aspects of the high-resolution fMRI technique and the sensitivity and spatial specificity of BOLD-based fMRI. We demonstrate applications of high-resolution fMRI in studying the human visual pathway from the lateral geniculate nucleus in the thalamus to the ocular dominance columns in the primary visual cortex. Most results were obtained at very-high-magnetic fields (3.0 and 4.0 Tesla). They reveal that high resolution fMRI at very-high-magnetic field is promising for functional mapping of brain organization from large cortical networks, small nuclei, and even to cellular layer structures. PMID- 10389674 TI - Very-high-field magnetic resonance imaging: instrumentation and safety issues. AB - Because of their advantage in terms of signal-to-noise ratio, high-field magnetic resonance imaging systems have become favored in the last few years for functional magnetic resonance imaging (fMRI) applications. In many ways the conceptual development of these high-field scanners has involved more-or-less straightforward extensions of practices at lower field strengths. However, in other ways specific engineering challenges have been encountered and largely overcome in the quest for scanners capable of realizing the advantages of high field systems. An understanding of the technical trade-offs that can be made in terms of hardware performance is useful in deciding on the optimum system for a given fMRI application. In this article the technical issues surrounding high field scanning are reviewed in the context of a typical brain mapping protocol. In addition there is a discussion of the safety issues related to the use of these systems. PMID- 10389675 TI - Diabetic gastropathy: gastric neuromuscular dysfunction in diabetes mellitus: a review of symptoms, pathophysiology, and treatment. AB - Diabetic gastropathy is a term that encompasses a number of neuromuscular dysfunctions of the stomach, including abnormalities of gastric contractility, tone, and myoelectrical activity in patients with diabetes. These abnormalities range from tachygastrias to antral hypomotility and frank gastroparesis. Diabetic gastropathies may be acutely produced during hyperglycemia. Symptoms of chronic diabetic gastropathy include chronic nausea, vague epigastric discomfort, postprandial fullness, early satiety, and vomiting. Because these symptoms are nonspecific, other disorders such as mechanical obstruction of the gastrointestinal tract, gastroesophageal reflux disease, cholecystitis, pancreatitis, mesenteric ischemia, and drug effects should be considered. Neuromuscular abnormalities of the stomach may be assessed noninvasively with gastric emptying tests, electrogastrography, and ultrasound. Gastrokinetic agents such as metoclopramide, cisapride, domperidone, and erythromycin increase fundic or antral contractions and/or eradicate gastric dysrhythmias. Diet and glucose control also are important in the management of diabetic gastropathy. As the pathophysiology of diabetic gastropathy is better understood, more specific and improved treatments will evolve. PMID- 10389676 TI - Gastric emptying of indigestible versus digestible oils and solid fats in normal humans. AB - Recent scintigraphic studies indicate that lipolytic products in the small intestine do not inhibit gastric emptying of fat as potently as previously suggested by studies that compared a liquid indigestible oil with a solid digestible fat. The older studies left open the confounding possibility that solid fats emptied differently than liquid oil. We studied eight normal subjects who ingested four meals in which fat was (1) liquid, digestible Crisco oil, (2) liquid, indigestible sucrose polyester oil, (3) digestible, solid Crisco, and (4) indigestible, solid olestra. Fats were labeled with iodine-123, and their gastric emptying was followed with a gamma camera. Indigestible fats (whether liquid or solid) emptied consistently faster than digestible fats (P < 0.005), although differences were small. Solid fats emptied about as rapidly as oils in the first hour; but more slowly thereafter (P < 0.01). A comparison of present scintigraphic with older studies suggested that solid fats were not well tracked by duodenal, marker-perfusion techniques, which misled previous investigators. PMID- 10389677 TI - Intragastric capsaicin stimulates motility of upper gut and proximal colon via distinct pathways in conscious dogs. AB - The aim of the present study was to investigate the effect and mechanism of action of intragastric and intraduodenal capsaicin on gastrointestinal motility. Five mongrel dogs were equipped with eight strain gauge force transducers on the stomach, small intestine, and proximal colon. In the interdigestive state, capsaicin was administered into the gastric or duodenal lumen. The effects of atropine, hexamethonium, ondansetron, and FK888 on capsaicin-induced contractions were studied. Intragastric capsaicin induced contractions within 15 min in the gastric antrum, duodenum, proximal jejunum, and proximal colon. These stimulatory effects were inhibited by atropine at all sites; by hexamethonium in the small intestine and colon; by ondansetron in the antrum, duodenum, and colon; and by FK888 in the antrum and colon, respectively. Intraduodenal capsaicin had no effect on contractility. Stimulation of afferent fibers by capsaicin in the stomach but not in the duodenum augments contractile activity in local and distant regions of the gut via distinct pathways. PMID- 10389678 TI - A comparative study of esophageal and anorectal motility in myotonic dystrophy. AB - Myotonic dystrophy may be associated with visceral abnormalities involving smooth muscle, the pathogenesis of which is not clear. Our aim was to evaluate the involvement of smooth and striated muscles at both ends of the gastrointestinal tract. Esophageal and anorectal manometric studies were performed in 13 patients and healthy controls. There was a correlation between: (1) the resting pressure in the upper esophageal sphincter and in the lower anal canal, (2) the amplitude and the coordination of contraction primary waves in the proximal and in the distal esophagus, and (3) the resting pressure in the higher anal canal and in the lower one. These results suggest that both ends of the gastrointestinal tract are disturbed in a similar fashion, both quantitatively and qualitatively and that there is a relationship between smooth and striated visceral muscle involvement in myotonic dystrophy. PMID- 10389679 TI - Effect of EM574 on postprandial pancreaticobiliary secretion, gastric motor activity, and emptying in conscious dogs. AB - EM574, an erythromycin derivative and a potent motilin receptor agonist, is now under clinical trial as a gastroprokinetic drug. The aim of this study was to estimate the effect of EM574 on postprandial pancreaticobiliary secretion, gastric motor activity, and emptying in conscious dogs. Five mongrel dogs were prepared. Indwelling cannulas for both infusion of phenolsulfonphthalein and aspiration of luminal samples were inserted into the proximal and distal duodenum, respectively. EM574 (3-30 microg/kg) was given intraduodenally through the indwelling distal duodenal cannula at the start of feeding. Postprandial pancreatic and biliary secretions were assessed by measuring the outputs of amylase and bile acid into the duodenum, respectively. Gastric motor and emptying activity were measured by means of a force transducer method and our own freeze drying method, respectively. One hundred grams of a freeze-dried standard meal was given as a solid marker after being mixed with 100 ml of normal saline containing 15 g of polyethylene glycol as a liquid marker. EM574 at doses of 10 and 30 microg/kg significantly increased the mean integrated postprandial amylase output into the duodenum, but the mean integrated postprandial bile acid output was not significantly increased. EM574 increased postprandial gastric antral motor activity dose-dependently. EM574 at doses of 10 and 30 microg/kg significantly accelerated gastric emptying of liquids and solids, respectively. EM574 enhances gastric antral motor activity and accelerates gastric emptying of solids and liquids with a concomitant increase in postprandial pancreatic amylase, but not bile acid, output in normal dogs. PMID- 10389680 TI - Effect of tramadol and morphine on pain and gastrointestinal motor function in patients with chronic pancreatitis. AB - Tramadol and morphine were compared for treatment of severe chronic pancreatitis pain and their interaction with gut motor function. Oral tramadol or morphine doses were titrated double-blinded and randomized for five days in 25 patients and pain, side effects, bowel function, orocecal and colonic transit, anal resting pressure, and rectal distension thresholds were measured. Pain intensities (mean+/-SD, 0 = none, 100 = unbearable) before treatment and on day 4 were 75+/-19 and 8+/-13 with tramadol (P < 0.001), and 65+/-21 and 5+/-6 with morphine (P < 0.001). On day 4, 67% of patients with tramadol and 20% with morphine rated their analgesia as excellent (P < 0.001) with mean respective doses of 840 mg (range: 80-1920) and 238 mg (20-1125). Orocecal transit was unchanged after five days of tramadol, but increased with morphine (P < 0.05). More patients had prolonged colonic transit times with morphine by day 5 (P < 0.05). Rectal distension threshold pressures increased only with tramadol (P < 0.01). It is concluded tramadol and morphine are potent analgesics in severe chronic pancreatitis pain when individually titrated. Tramadol interfered significantly less with gastrointestinal function and was more often rated as an excellent analgesic than morphine. PMID- 10389681 TI - EGF and TGF-beta1 gene expression in chronically rejecting small bowel transplants. AB - Long-term survival of small bowel transplants is hampered by chronic rejection. Epidermal growth factor (EGF) and transforming growth factor beta (TGF-beta) have opposing, regulatory roles in normal intestinal physiology and may be involved in the pathogenesis of chronic intestinal rejection. Our aim was to investigate the expression of EGF and TGF-beta1 in chronically rejecting small bowel transplants. Orthotopic small bowel transplantation was performed in the allogeneic DA-to-AS rat combination; Cyclosporin was administered temporarily to prevent acute rejection. Controls were DA isografts and normal DA rats. PreproEGF and TGF-beta1 gene expression was evaluated by northern blot analysis of the ileum RNA and standardized against glyceraldehyde-3-phosphate-dehydrogenase expression. Allografts demonstrated functional impairment and histological features of chronic rejection, whereas isografts appeared normal. Allografts demonstrated a significant reduction of EGF mRNA when compared to DA isografts. No significant changes were detected in TGF-beta1 expression in either allogeneic or syngeneic grafts. In conclusion, this study demonstrates reduced preproEGF and preserved TGF-beta1 gene expression in chronically rejecting small bowel transplants. PMID- 10389682 TI - Clinical relevance of acute pancreatitis in allogeneic hemopoietic stem cell (bone marrow or peripheral blood) transplants. AB - This study investigated the clinical relevance of acute pancreatitis in allogeneic hemopoietic stem cell (bone marrow or peripheral blood) transplants (BMT). We studied 26 patients undergoing BMT. The preparative regimen was busulfan and cyclophosphamide in 17 patients and total body irradiation and cyclophosphamide in 9 patients. Graft-versus-host disease (GVHD) prophylaxis consisted of cyclosporin A and short-term methotrexate in all 26 patients. The pancreas was studied using amylase and lipase serum levels, abdominal contrast enhanced tomography, and/or ultrasound. Clinical and laboratory signs of acute pancreatitis were found in two patients with acute hepatointestinal GVHD, and in one patient with acute hepatic GVHD and cytomegalovirus infection. This patient died of multiorgan failure, with interstitial acute pancreatitis at autopsy; the other two patients recovered with general supportive care and GVHD therapy. We suggest that in the patients with complications after BMT, particularly acute hepatic/hepatointestinal GVHD, and cytomegalovirus infection, the possibility of acute pancreatitis should be considered. PMID- 10389683 TI - Splenic peliosis: a rare complication following liver transplantation. AB - In this article, we report a rare case of isolated splenic peliosis in an individual who had recently undergone liver transplantation. The disorder had remained clinically and radiologically undiagnosed until he suffered a traumatic rupture of the affected organ. The relevant literature on this topic is briefly reviewed. PMID- 10389684 TI - Gastric acid blockade with omeprazole promotes gastric carcinogenesis induced by duodenogastric reflux. AB - Duodenogastric reflux (DGR) in rats causes growth stimulation of the foregut mucosa that is potentiated by gastric acid blockade. It was the aim of this study to investigate if DGR with gastric acid blockade has a higher incidence of carcinomas of the foregut than DGR alone. DGR was induced in 40 Sprague-Dawley rats using a split gastroenterostomy. A cardiomyotomy was performed across the gastroesophageal junction, inducing reflux into the esophagus. Twenty of these rats received omeprazole postoperatively. After one year 18 rats (90%) with DGR + omeprazole treatment and 7 rats (35%) with DGR alone developed adenocarcinoma of the stomach (P < 0.05). None of the rats developed esophageal cancer, but esophageal mucosal hyperplasia was more pronounced in rats receiving omeprazole. Control rats, treated with omeprazole, did not develop carcinomas of the foregut. In conclusion, gastric acid blockade enhanced DGR-induced carcinogenesis of the stomach and promotes growth stimulation of the esophageal mucosa. PMID- 10389685 TI - Overexpression of epidermal growth factor receptor in Peutz-Jeghers syndrome. AB - Peutz-Jeghers syndrome is characterized by gastrointestinal hamartomatous polyposis, mucocutaneous pigmentation, and a predisposition to cancer. The etiology of this syndrome is unknown. We investigated the expression of epidermal growth factor receptor (EGFr), transforming growth factor-alpha (TGF-alpha), transforming growth factor-beta1 (TGF-beta1) and transforming growth factor-beta receptor (TGF-beta RII) between normal and Peutz-Jeghers small bowel tissues. In addition, immunoprecipitation by phosphotyrosine antibodies followed by EGFr western blotting was measured and compared between a Peutz-Jeghers hamartoma and normal duodenal tissue. EGFr expression was increased 2.5-fold in normal and hamartomatous tissue of Peutz-Jeghers patients compared to normal small bowel tissue. In Peutz-Jeghers tissues, the major EGFr immunoreactive band was increased size from 170 to approximately 200 kDa. Using an antibody specific for activated EGFr, this larger size band was predominant in Peutz-Jeghers tissue. Immunoprecipitation of a hamartoma by a phosphotyrosine specific antibody followed by western blotting for EGFr demonstrated this 200-kDa band. Expression of TGF-alpha, TGF-beta1, TGF-beta1 RII was not significantly different between normal and Peutz-Jeghers tissues. In conclusion, EGFr was overexpressed in normal and hamartomatous small bowel tissue of Peutz-Jeghers patients, which suggests that EGFr in Peutz-Jeghers tissue is persistently activated or highly stimulated by endogenous ligands and also suggests a possible role for EGFr in the pathogenesis of Peutz-Jeghers syndrome. PMID- 10389686 TI - Serum levels of pancreatitis-associated protein in digestive diseases with special reference to gastrointestinal cancers. AB - The serum levels of pancreatitis-associated protein (PAP) were measured in 196 patients with digestive diseases and 15 healthy subjects by an enzyme-linked immunosorbent assay. The serum PAP levels were significantly elevated in the patients with gastric, colorectal, biliary tract, hepatocellular, or pancreatic cancers compared with the healthy subjects. After curative resection of the tumor, serum PAP levels were significantly decreased. The serum PAP levels were not related to clinicopathological factors except for the tumor size of pancreatic cancer. There were some cases of PAP-positive and carcinoembryonic antigen (CEA) or carbohydrate antigen (CA) 19-9 -negative gastric and colorectal cancers. The serum PAP levels were also significantly elevated in the patients with acute pancreatitis compared with those in not only the healthy subjects but also the patients with chronic pancreatitis. The peak PAP levels were significantly correlated with the severity of acute pancreatitis and reflected the clinical healing of the disease. The peak of serum PAP was significantly delayed compared with those of other pancreatic enzymes. These results suggest that the increase of serum PAP levels in patients with gastrointestinal cancers reflects an ectopic expression of PAP in cancer cells and that increased serum levels of PAP in acute pancreatitis are correlated with the disease severity and are prolonged than those of other pancreatic markers. PMID- 10389687 TI - Long-term survival after diagnosis of hepatic metastatic VIPoma: report of two cases with disparate courses and review of therapeutic options. AB - This report describes two patients with pancreatic cholera caused by vasoactive intestinal polypeptide (VIP)-producing tumors, which originated in the pancreas and showed metastases in both hepatic lobes at time of diagnosis. However, the two tumors displayed remarkably disparate clinical courses. Due to the protracted but progressive course over more than 10 years, a multifaceted therapeutic approach was performed to control symptoms and to improve quality of life. The long-acting somatostatin analog octreotide was the most effective treatment for relieving symptoms and correcting fluid and electrolytes disturbances. The effects of complementary treatments, including systemic chemotherapy and hyperselective chemoembolization, as well as concurrent application of octreotide and prednisolone or interferon with respect to clinical symptoms, VIP levels, and tumor growth are reviewed. Our experience, although small, emphasizes the need for an expert, well-planned, adaptive, and multidisciplinary approach in the care of these complex patients. PMID- 10389688 TI - Predictive factors and prevalence of follicular gastritis in adults with peptic ulcer and nonulcer dyspepsia. AB - Follicular gastritis is an important histological entity, because it may progress to overt gastric MALT lymphoma. However, there is no universal agreement on whether there is any correlation of follicular gastritis with histological features of the antral mucosa or on the prevalence of follicular gastritis. To shed further light on these issues, we studied antral biopsies obtained from 735 adult patients, who had participated in six consecutive clinical trials. They included 348 patients with duodenal ulcer, 82 with gastric ulcer, and 305 with nonulcer dyspepsia. The Sydney classification system of gastritis was used, using a score of 0-3 to grade degree and activity of inflammation, gland atrophy, intestinal metaplasia, and H. pylori colonization density. Follicular gastritis was defined as prominent lymphoid follicles with no lymphoepithelial lesion. None of the H. pylori-negative patients (N = 159) had follicular gastritis. Among H. pylori-positive patients, 80/340 (23.5%) with duodenal ulcer, 5/77 (6.5%) with gastric ulcer, and 20/159 (12.6%) with nonulcer dyspepsia had follicular gastritis (P < 0.001). Multivariate discriminant analysis selected the following four significant predictor variables for follicular gastritis (Wilks lambda = 0.91, chi2 = 70.6, df = 4, P < 0.001): gastritis sum score, atrophic gastritis, age of the patient, and disease. The prevalence of follicular gastritis was linearly correlated (gamma = 24.55 - 0.98chi, r = -0.62, F1,11 = 6.12, P = 0.03) with the age groups of the 576 H. pylori-positive patients studied. In conclusion, follicular gastritis is highly correlated with H. pylori-caused severe, active gastritis. It is mostly prevalent in the young H. pylori-infected patients with duodenal ulcer. PMID- 10389689 TI - High prevalence of Helicobacter pylori infection in shepherds. AB - It has been suggested that Helicobacter pylori infection may, in some instances, be a zoonosis. The aim of this study was to evaluate the prevalence of H. pylori infection in Sardinian shepherds and their families in relation to exposure to sheep and sheep dogs. Sardinian shepherds and a control group of blood donors completed detailed questionnaires regarding demographics, childhood and current economic status, and the presence of symptoms related to the upper gastrointestinal tract. H. pylori status was determined by a sensitive ELISA for anti-H. pylori IgG and by western blot for anti-CagA IgG. A subgroup of shepherds had upper gastrointestinal endoscopy with biopsy to assess the severity of the gastritis. H. pylori infection in Sardinian shepherds approached 100% and was positively related to animal contact (98% of shepherds, 73% of family members without regular direct animal contact compared to 43% of blood donors) (P < 0.001). Importantly, the family members shared the same childhood with the shepherds but choose different careers (e.g., teachers, nurses, business) and did not have regular contact with sheep. In conclusion, the prevalence of H. pylori infection in Sardinian shepherds is among the highest in the world and is associated with direct contact with sheep and sheep dogs. These results suggest that the cycle of H. pylori infection might, in certain circumstances, include phases in the environment, animals (sheep or dogs) and human beings. PMID- 10389690 TI - Examination of tissue distribution of Helicobacter pylori within columnar-lined esophagus. AB - H. pylori may colonize columnar-lined esophagus, although an etiologic role in esophageal adenocarcinoma is unproven. H. pylori can adhere to intestinal metaplasia in the stomach. This study was designed to examine if H. pylori adheres to specialized intestinal metaplasia in columnar-lined esophagus. Esophageal biopsies from patients with columnar-lined esophagus were reviewed. Patients with only gastric metaplasia were excluded. Sections with specialized intestinal metaplasia in at least one third of at least one gland were recut, stained using the Giemsa stain, and reexamined by two independent pathologists using strict criteria for adherence by H. pylori. The 209 esophageal biopsies with adequate specialized intestinal metaplasia from 58 patients were examined: H. pylori was only seen on gastric metaplasia in three patients-and never on specialized intestinal metaplasia. Within the esophagus, H. pylori adheres only to gastric metaplasia, which is not considered premalignant for esophageal adenocarcinoma. PMID- 10389691 TI - Detection of Helicobacter pylori organisms by Hp-fast in children. AB - Hp-fast is a new rapid urease test (RUT) that has not been evaluated in children. The aim of the study was to prospectively compare the Hp-fast test to the CLOtest in children. Children with gastrointestinal symptoms who undergo diagnostic upper endoscopy were prospectively enrolled to the study. Antral gastric biopsies were evaluated for histology and for CLO-test and Hp-fast. Results were then compared to histology. Of the 94 children who participated, gastritis was found in 38 (40%), of whom 16 (42%) had associated H. pylori organisms. In two children, H. pylori organisms were identified without gastritis. The concordance between both RUT tests was 98%. A significant correlation was found between RUT results and histological factors or serology. The accuracy rate of both RUT increased significantly when different gold standards were utilized to detect Hp infection in children. The best correlation was found when histology and serology were considered as the gold standard for the diagnosis of H. pylori infection in children (sensitivity: 100% compared to 43-80% with other standards, respectively). In conclusion, the Hp-fast test result is comparable to CLOtest, but neither alone is sufficient to establish the diagnosis of Hp infection in children. PMID- 10389692 TI - Use of chopsticks for eating and Helicobacter pylori infection. AB - Epidemiological data suggests that ethnic groups using chopsticks for eating have a higher prevalence of H. pylori infection. This study investigated the carriage of H. pylori in chopsticks after eating. Used chopsticks and saliva were collected from asymptomatic individuals whose H. pylori status was determined by [13C]urea breath test and serology. Both the saliva specimens and chopsticks were cultured and processed by polymerase chain reaction (PCR) for the detection of H. pylori. Furthermore, chopsticks used by hospital staff in the cafeteria were pooled for the detection of H. pylori by bacteriologic culture and PCR. Sixty nine volunteers were recruited in the first study and 45 (65%) were diagnosed to have H. pylori infection. While all cultures were negative, H. pylori was detected by PCR in the saliva from 15 (33%) infected subjects and in the chopsticks from one (2%). Among the 12 sets of pooled chopstick-washing studied, H. pylori was detected by PCR in two sets. This study showed that H. pylori was rarely detected in chopsticks after eating and hence, the risk of contracting this infection via the use of chopsticks is low. PMID- 10389694 TI - Duodenal obstruction caused by Strongyloides stercoralis enteritis in an HTLV-1 infected host. PMID- 10389693 TI - Bile reflux due to disturbed gastric movement is a cause of spontaneous gastric ulcer in W/Wv mice. AB - c-Kit is a receptor tyrosine kinase, and it is encoded by the mouse W locus. Mutant W/Wv mice develop spontaneous gastric antral ulcers. The aim of the present study was to investigate the pathogenesis of these gastric ulcers and to examine the effects of two antiulcer drugs; a proton pump inhibitor (2{[4-(3 methoxypropoxy)-3-methylpyridine-2-yl]methyl-sulfinyl}-1H -benzimidazole sodium salt, rabeprazole) and a mucosal protective drug (geranylgeranylacetone, GGA), on the gastric ulcers. The inhibition of the gastric acid secretion by rabeprazole (30 mg/kg body weight, subcutaneous injection once a day for six weeks) significantly increased the gastric ulcer formation compared to the controls. In contrast, the GGA treatment (100 mg/kg body weight, oral administration for six weeks) significantly inhibited the ulcer formation. Bile reflux was seen in these mutant mice, and they showed no cyclic intense contractions in the gastric antrum. These results suggest that bile reflux due to the disturbance of gastric antral movement is a cause of the spontaneous gastric ulcers in W/Wv mice. PMID- 10389695 TI - Fatal ulcerative panenteritis following colectomy in a patient with ulcerative colitis. AB - A 37-year-old man, previously submitted to colectomy for ulcerative pancolitis unresponsive to medical therapy, presented with nausea, vomiting, epigastric pain, and bloody diarrhea. An upper gastrointestinal endoscopy revealed mucosal friability, petechiae, and erosions throughout the duodenum, whereas prestomal ileum showed large ulcers and pseudopolyps. Histologically, a dense inflammation chiefly composed of lymphocytes and plasma cells with few neutrophils was detected. No bacteria, protozoa, and fungi could be detected. Despite intensive care, intra-1194 venous antibiotics and steroids, the patient died of diffuse intravascular coagulation and multiorgan failure. At post-mortem examination severe ulcerative lesions were observed scattered throughout the duodenum up to the distal ileum. The dramatic clinical presentation with fatal outcome, the widespread ulcers throughout the intestine, and the histological picture are peculiar features in our patient which can not be ascribed to any type of the ulcerative jejunoenteritis so far reported. Patients with pancolitis and diffuse ileal involvement do not necessarily have Crohn's disease but rather may have ulcerative colitis. PMID- 10389696 TI - Vascular endothelial growth factor (VEGF) in Crohn's disease: increased production by peripheral blood mononuclear cells and decreased VEGF165 labeling of peripheral CD14+ monocytes. AB - Recently, increased serum levels of vascular endothelial growth factor (VEGF) have been shown in patients with inflammatory bowel disease. The origins of the circulating VEGF are still not described. Monocytes play an important role in the inflammatory process. VEGF binding to monocytes mediates monocyte recruitment and activation. The present study investigates the VEGF production of peripheral blood mononuclear cells and the ability of peripheral monocytes to bind VEGF165 in patients with Crohn's disease. Nineteen patients with Crohn's disease and 10 healthy volunteers were studied. VEGF165 labeling of CD14+ monocytes was measured using two-color flow cytometry. Density of VEGF labeling was expressed as the mean fluorescence intensity (MFI). Furthermore, VEGF levels were determined in culture supernatants of unstimulated peripheral blood mononuclear cells. VEGF in culture supernatants was measured using a solid-phase enzyme-linked immunosorbent assay. There was a significantly decreased VEGF165 labeling of monocytes of patients with active Crohn's disease (MFI: 369.9+/-121.6, N = 7, P < 0.002) compared to patients with inactive disease (MFI: 457.7+/-74.5, N = 6) and healthy controls (MFI: 542.9+/-96.2, N = 10). Unstimulated peripheral blood mononuclear cells of patients with active Crohn's disease produced significantly higher amounts of VEGF (1142.6+/-483.9 pg/ml, N = 12, P < 0.001) compared with peripheral blood mononuclear cells of healthy volunteers (113.4+/-101.8 pg/ml, N = 10). VEGF production by peripheral blood mononuclear cells of patients with active disease was significantly increased compared to patients with quiescent disease (261.6+/-254.8 pg/ml, N = 7, P < 0.001). In conclusion, our data describe peripheral blood mononuclear cells as one of the origins of the elevated VEGF serum levels in patients with active Crohn's disease. Furthermore, a decrease in VEGF165 binding sites on peripheral monocytes of patients with active Crohn's disease has been shown. The study underlines the important role of VEGF in Crohn's disease. PMID- 10389697 TI - Serum antibodies to Mycobacterium paratuberculosis in patients with Crohn's disease. AB - Mycobacterium paratuberculosis has been suggested as a causative organism of Crohn's disease. Despite a long-term debate to prove this possibility, the role of this bacteria in the pathogenesis of Crohn's disease is still a subject of controversy. In the present study, serum antibodies (IgG, IgA, and IgM) to the protoplasmic antigen of M. paratuberculosis were quantified in patients with Crohn's disease and in control subjects by using an enzyme-linked immunosorbent assay whose specificity was increased by preabsorbing the sera with cell extracts of Mycobacterium phlei. As compared to normal controls (1/20; 0.062+/-0.022), a significant difference was seen in the antibody-positive prevalence rate and mean values of the serum IgG titer in patients with Crohn's disease (5/13; 0.102+/ 0.039) (P < 0.05), but not in patients with ulcerative colitis (2/20; 0.065+/ 0.035) and tuberculosis (0/4; 0.053+/-0.008). No significant differences were seen in the antibody-positive prevalence rate and mean values of the serum IgA and IgM titers among the four study groups. These results indicate the unique immune response to M. paratuberculosis in patients with Crohn's disease, suggesting that this organism may play some role in the pathogenesis of Crohn's disease. PMID- 10389698 TI - Decreased corticosensitivity in quiescent Crohn's disease: an ex vivo study using whole blood cell cultures. AB - Corticosensitivity influences the degree and the duration of an inflammatory reaction by altering target cell responses to endogenous and/or exogenous glucocorticoids. Indeed, different clinical responses to glucocorticoids have been observed among patients with Crohn's disease, suggesting different degrees of corticosensitivity in these subjects. The purpose of this study was to compare the corticosensitivity of patients with quiescent Crohn's disease to that of healthy subjects (HS). Nineteen patients with quiescent Crohn's disease and 14 HS were studied; all patients were steroid-free for at least six months; 7 of the 19 were corticosteroid-dependent (CSD) and treated with nonglucocorticoid immunosuppressants at the time of the study. Corticosensitivity was measured by the inhibition of LPS-induced cytokine secretion in whole blood cell cultures treated with increasing concentrations (10(-9) to 10(-6) M) of dexamethasone. Tumor-necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), and interleukin-1 beta (IL-1 beta) were measured using specific immunoassays. Crohn's disease patients had a markedly decreased dexamethasone-mediated inhibition of TNF-alpha (P < 0.01), IL-6 (P < 0.001), and IL-1 beta (P < 0.01) compared to healthy subjects, with a shift of the dexamethasone dose-response curve to the right. No significant differences in the basal and LPS-stimulated secretion of the three cytokines were observed between CSD and non-CSD patients, and both subgroups of patients had similar degrees of dexamethasone-mediated cytokine inhibition. We conclude that patients with Crohn's disease have a significant decrease in the corticosensitivity of their leukocytes. This may be related to a specific genetic/constitutional background and/or could be acquired, due to inflammation related endocrine and/or immune factors. PMID- 10389699 TI - Vegetables, high nitrate foods, increased breath nitrous oxide. AB - We have demonstrated nitrous oxide (N2O), a metabolite of the reduction of nitrate (NO(3)-) by microflora, in exhaled air. The purpose of this study is to examine the effect of ingesting vegetables, which contain high levels of NO(3)-, on breath N2O levels. We measured breath N2O in six healthy subjects aged 20-41 years at 15-min intervals after the following: (1) no ingestion; (2) ingestion of 180 g of vegetable juice; and (3) ingestion of 50 g of lettuce. N2O levels were measured with an infrared-photoacoustic (IR-PAS) analyzer. N2O was detectable in all subjects regardless of treatment protocol. Lettuce and vegetable juice significantly increased breath N2O levels. Total excretions in breath N2O were significantly higher in the lettuce group (P = 0.028) and the vegetable juice group (P = 0.046) than controls. Breath N2O increases after vegetable ingestion, probably due to denitrification of NO(3)- by normal microflora in the intestinal tract. PMID- 10389700 TI - Rifaximin in patients with moderate or severe ulcerative colitis refractory to steroid-treatment: a double-blind, placebo-controlled trial. PMID- 10389701 TI - Rapid estimation of creatinine clearances in patients with liver dysfunction. AB - Different methods are available for rapid assessment of renal function using the patient's serum creatinine concentration and body weight without obtaining urine collection over 24 hr. However, the reliability of these methods in patients with liver diseases has not been established. The purpose of this study was to determine the accuracy and precision of the estimated creatinine clearances obtained by the methods of Cockcroft-Gault, Jelliffe, Mawer, and Siersbaek Nielsen in patients with liver diseases who have different degrees of renal function. Creatinine clearances obtained from 24-hr urine collection were used as the standard. The different methods for rapid renal function estimation had limited accuracy and reliability in patients with severe liver dysfunction (Child Pugh class C) and also in those with creatinine clearances of less than 60 ml/min. Creatinine clearances were overestimated by about 40-100%. Using lean body weights, instead of total body weights, reduced the prediction errors. In patients with mild liver dysfunction (Child-Pugh class A), all four estimation methods provided reasonable estimation of the creatinine clearances. PMID- 10389702 TI - Hypothyroidism protects rat liver from acetaminophen hepatotoxicity. AB - Recent data from animal studies suggest that induced hypothyroidism inhibits the development of liver injury in several animal models, including liver cirrhosis and fulminant hepatic failure in rats, and immune-mediated acute liver injury in mice. The aim of the present study was to determine whether hypothyroidism would likewise prevent acetaminophen-induced hepatic damage in rats. Liver damage was induced by acetaminophen (2 g/kg) administered by gavage to fasting rats as a single dose. Hypothyroidism was induced by methimazole, propylthiouracil, or surgical thyroidectomy and confirmed by elevated serum levels of TSH. Hypothyroidism significantly inhibited acetaminophen-induced liver damage as manifested by the decreased serum levels of liver enzymes, malondialdehyde and blood ammonia, as well as by the higher hepatic glutathione content, in all three groups of hypothyroid rats compared to euthyroid controls (P < 0.01). Histopathologic analysis showed significantly less liver necrosis and inflammation in the acetaminophen-treated hypothyroid rats. Oxygen extraction, measured in isolated perfused rat liver preparation, was also reduced in the hypothyroid livers to 42+/-8% compared to 81+/-14% of controls (P < 0.01). However, the expression of CYP2E1 in the livers of hypothyroid rats, as measured by western blot analysis, was not decreased compared to control rats. These results suggest that induced hypothyroidism, regardless of the mode of induction, protects rat liver from acetaminophen hepatotoxicity. This effect may be related to hypometabolism of liver cells, but the exact mechanism needs further clarification. PMID- 10389703 TI - Interaction of alcohol and hepatitis C virus infection on severity of liver disease. AB - Chronic alcoholics have a high prevalence of hepatitis C virus (HCV) infection. The present study was carried out to examine the association between HCV infection and alcohol abuse, and the influence of these factors on the severity of liver disease. Patients with history of heavy alcohol abuse (> or = 80 g of ethanol per day for > or = 5 years) were analyzed with respect to the amount of alcohol use, clinical evidence of liver disease, and laboratory tests. One hundred ninety-nine patients, 137 HCV positive and 62 HCV negative were included in the study. HCV-infected subjects had liver disease for a longer duration (P < 0.0001) and had higher incidence of symptoms of hepatic decompensation in the past compared to uninfected alcoholics. Several differences were noted between the two groups at the time of presentation to the hospital. Alcoholics with HCV infection had lower daily alcohol consumption (P < 0.001), were abstinent for a longer duration (P < 0.02) and had lower lifetime use of ethanol (P < 0.005) compared to HCV-negative subjects. Assessment of liver tests showed greater derangement in uninfected alcoholics compared to HCV-positive subjects. The present study shows that HCV-infected chronic alcoholics have lower alcohol consumption and, perhaps as a consequence, have less severe liver disease compared to HCV-negative individuals. These findings suggest that in chronic alcoholics, despite the presence of HCV infection, the severity of liver damage is related to the amount of alcohol consumption. PMID- 10389704 TI - Diagnosis of hepatopulmonary syndrome with contrast transesophageal echocardiography: advantages over contrast transthoracic echocardiography. AB - The aim of this study was to study the prevalence of hepatopulmonary syndrome (HPS) in cirrhotic patients, comparing the results obtained using contrast transthoracic echocardiography (CTTE) and contrast transesophageal echocardiography (CTEE) in the demonstration and grading of pulmonary vasodilatation (PV). We also analyzed the correlation between gas-exchange abnormalities and PV when it was demonstrated with both techniques. The prevalence of PV and HPS with CTEE in the 88 cirrhotic patients was 28% and 22%, respectively, and with CTEE, 42% (P < 0.05) (middle PV: 35% and significant PV: 7%) and 30% (P < 0.05), respectively. Fifty-six percent of patients diagnosed with PV and with CTTE presented with hypoxemia as did 49% using CTEE (83% of patients with significant PV had hypoxaemia). PaCO2 and diffusing capacity of CO were significantly more decreased in patients with PV than in patients without PV when CTEE was employed. We conclude that CTEE is superior to CTTE in the diagnosis and grading of PV in the diagnosis of HPS in cirrhotic patients, being more sensitive and presenting a better correlation with gas-exchange abnormalities. Given its high sensitivity, CTEE should be carried out in all patients with suspicion of HPS and normal or uncertain CTTE. PMID- 10389705 TI - Progressive liver functional impairment is associated with an increase in AST/ALT ratio. AB - The ratio of serum aspartate aminotransferase to alanine aminotransferase (AST/ALT ratio) has been proposed as a noninvasive method of assessing liver fibrosis and cirrhosis. Our aims were to confirm the usefulness of the AST/ALT ratio in diagnosing cirrhosis noninvasively as well as to verify the existence of a relationship between the ratio and liver functional impairment. In all, 348 patients (177 with chronic hepatitis, 171 with cirrhosis) were retrospectively evaluated and the AST/ALT ratio was related to monoethyl glycine xylidide (MEGX) formation. Moreover, in a subgroup of 54 patients we analyzed the relationships among the AST/ALT ratio and indocyanine green clearance and half-life. The AST/ALT ratio was able to separate patients with mild fibrosis from those with severe fibrosis and cirrhosis. The AST/ALT ratio, MEGX, prothrombin activity, and platelet count were selected by multivariate analysis as variables associated with cirrhosis. The AST/ALT ratio showed significant correlations both with MEGX formation and with indocyanine green clearance and half-life. The alterations of indocyanine green kinetics, which depend upon liver blood flow and uptake, were likely due to progressive fibrosis. These findings might partially explain the increase in the AST/ALT ratio as disease progresses. PMID- 10389706 TI - Impaired adaptive cytoprotection to ethanol-induced damage in gastric mucosa of portal hypertensive rats. AB - Portal hypertension predisposes gastric mucosa to increased damage by noxious agents. Adaptive cytoprotection has not been studied in portal hypertensive gastric mucosa. We evaluated adaptive cytoprotection in the gastric mucosa of portal hypertensive rats by exposure to ethanol. The injury index (percent gross lesions) was significantly higher in portal hypertensive rats than in sham operated rats. The ratio of adaptive cytoprotection, calculated as the degree of decrease in the injury index caused by pre-absolute-ethanol administration of 20% ethanol, was significantly impaired in portal hypertensive rats. Basal levels of gastric mucosal hexosamine were lower in portal hypertensive rats than in controls, and a blunted response to 20% ethanol was associated with portal hypertension. Nitric oxide inhibition (L-NAME, 5 mg/kg) reduced the ratio of adaptive cytoprotection in sham-operated but not in portal hypertensive rats. These results suggest that impaired adaptive cytoprotection in portal hypertensive gastric mucosa may be caused by blunted mucus production. PMID- 10389707 TI - Vascular endothelial growth factor levels in liver cirrhosis. PMID- 10389709 TI - Elevated amniotic fluid nucleosome levels in women with intra-amniotic infection. AB - OBJECTIVE: To compare amniotic fluid (AF) soluble nucleosome levels in pregnant women with and without intra-amniotic infection. METHODS: Amniocentesis was performed in 74 pregnant women with preterm contractions, labor, or premature rupture of membranes. Intra-amniotic infection was defined as a positive AF culture. Amniotic fluid tests for Gram stain, glucose, neutrophils, creatinine, pH, and specific gravity were performed. Amniotic fluid soluble nucleosome levels were determined by enzyme-linked immunosorbent assay and were normalized by AF creatinine levels. RESULTS: Twenty-eight patients had intra-amniotic infection and 46 did not. Amniotic fluid soluble nucleosome levels were significantly higher in pregnant women with intra-amniotic infection than in those without infection (48.1+/-21.3 compared with 0.0+/-0.0 U/mg creatinine; P = .005). The AF nucleosome levels were positively correlated with AF neutrophil counts and negatively correlated with AF glucose concentrations. CONCLUSION: Our data indicate that elevated AF nucleosome levels are associated with intra-amniotic infection and may have potential as a clinical marker to detect intra-amniotic infection. PMID- 10389708 TI - Amniotic fluid matrix metalloproteinase-9 levels in women with preterm labor and suspected intra-amniotic infection. AB - OBJECTIVE: To determine the accuracy of amniotic fluid (AF) matrix metalloproteinase-9 measurements for diagnosing intra-amniotic infection in women with preterm labor. METHODS: We performed amniocenteses in 44 women between 22 and 35 weeks' gestation who presented to our center with preterm labor and clinical suspicion of intra-amniotic infection. Each sample was analyzed by glucose measurement, Gram stain, and culture for aerobes, anaerobes, and mycoplasmas. We tested the AF for matrix metalloproteinase-9 using gelatin zymography and a commercial enzyme-linked immunosorbent assay (ELISA) system. We calculated accuracy and confidence intervals (CIs) for AF matrix metalloproteinase-9, glucose, and Gram stain for diagnosing intra-amniotic infection, using culture as the criterion standard. RESULTS: All patients who had matrix metalloproteinase-9 detectable by ELISA also demonstrated matrix metalloproteinase-9 by zymography. Six cases of intra-amniotic infection were confirmed by culture (prevalence 14%). The performance statistics of AF matrix metalloproteinase-9 for diagnosing intra-amniotic infection were: sensitivity 83% (95% CI 53, 99), specificity 95% (95% CI 88, 99), positive predictive value 71% (95% CI 37, 99), and negative predictive value 97% (95% CI 92, 99). Two women had false-positive results; one had gram-negative rods on the AF Gram stain and developed clinical signs and symptoms of chorioamnionitis several hours after amniocentesis and the other had a purulent vaginal discharge and an AF glucose level less than 15 mg/dL. Both delivered within 24 hours of amniocentesis. CONCLUSION: Measuring matrix metalloproteinase-9 in the AF appeared to be reliable for diagnosing intra-amniotic infection. An elevated matrix metalloproteinase-9 concentration in the AF at a preterm gestational age may portend imminent delivery regardless of microbiologic confirmation of intra amniotic infection. PMID- 10389710 TI - Prostaglandin E2 cervical ripening without subsequent induction of labor. AB - OBJECTIVE: To determine whether outpatient administration of intracervical prostaglandin (PG) E2 gel decreases the interval to delivery and duration of labor. METHODS: A randomized, double-blind, placebo-controlled trial compared the intracervical placement of 0.5 mg PGE2 gel with placebo in 61 pregnant women at 38 weeks' or greater gestation with Bishop scores less than 9. Transvaginal cervical length, fetal fibronectin, and Bishop score were assessed before gel placement. Subjects were then allowed to go into spontaneous labor unless an indication for induction developed. RESULTS: Thirty women were assigned to PGE2 and 31 to placebo. There were no significant demographic differences between the groups and there were no differences in cervical length, fetal fibronectin status, or Bishop scores. Fifteen women in the PGE2 group and five in the placebo group went into labor and delivered within the first 2 days after gel placement (P = .007). The median interval to delivery was significantly shorter in the PGE2 group, at 2.5 days, compared with placebo, at 7 days (P = .02). Nulliparas in the PGE2 group had a median interval to delivery of 2 days, compared with 7 days for nulliparas receiving placebo (P = .03). Active phases of labor were significantly shorter in the PGE2 group and for women with a negative fetal fibronectin test who received PGE2. CONCLUSION: Outpatient administration of intracervical PGE2 gel shortened intervals to delivery and shortened labor. PMID- 10389711 TI - Pregnancy in women with systemic sclerosis. AB - OBJECTIVE: To determine pregnancy outcomes in women with systemic sclerosis. METHODS: Women of childbearing age with systemic sclerosis seen at the University of Pittsburgh between 1987 and 1996 were observed prospectively. Pregnancy outcomes included abortion, miscarriage, preterm and term birth, and perinatal death. Complications of pregnancy and scleroderma were determined during and after pregnancy. RESULTS: Fifty-nine women with systemic sclerosis had 91 pregnancies during the 10-year period. No increase in the frequency of miscarriage was found, except in those with long-standing diffuse scleroderma. Preterm births occurred in 29% of pregnancies, and all but one of the infants survived. Symptoms related to scleroderma, particularly Raynaud phenomenon, improved during pregnancy, but esophageal reflux became worse. After pregnancy, some women with diffuse scleroderma had increased skin thickening. There were three cases of renal crisis during pregnancy, all in women with early diffuse scleroderma. Four women had five healthy infants while taking angiotensin converting-enzyme inhibitors. CONCLUSION: Women with systemic sclerosis can safely have healthy pregnancies. Those with early diffuse scleroderma should wait until their disease stabilizes before becoming pregnant to decrease the risk of renal crisis. High-risk pregnancy management should be standard for all scleroderma pregnancies because of the high frequency of premature births. PMID- 10389712 TI - Anal sphincter tears at vaginal delivery: risk factors and clinical outcome of primary repair. AB - OBJECTIVE: To determine risk factors for obstetric anal sphincter tears and to evaluate symptomatic outcome of primary repair. METHODS: Obstetric-procedure, maternal, and fetal data were registered in 845 consecutive vaginally delivered women. Risk factors for anal sphincter tears were calculated by multiple logistic regression. All 808 Swedish-speaking women who delivered vaginally were included in a questionnaire study regarding anal incontinence in relation to the delivery. Questionnaires were distributed within the first few days postpartum, and at 5 and 9 months postpartum. RESULTS: Six percent of the women had a clinically detected sphincter tear at delivery. Sphincter tears were associated with nulliparity (odds ratio [OR] 9.8, 95% confidence interval [CI] 3.6, 26.2), postmaturity (OR 2.5, 95% CI 1.0, 6.2), fundal pressure (OR 4.6 95% CI 2.3, 7.9), midline episiotomy (OR 5.5 95% CI 1.4,18.7), and fetal weight in intervals of 250 g (OR 1.3 95% CI 1.1, 1.6). Fifty-four percent of women with repaired sphincter tears suffered from fecal or gas incontinence or both at 5 months and 41% at 9 months. Most of the symptoms were infrequent and mild. CONCLUSION: Several risk factors for sphincter tear were identified. Sphincter tear at vaginal delivery is a serious complication, and it is frequently associated with anal incontinence. Special attention should be directed toward risk factors for this complication. Symptoms of anal incontinence should explicitly be sought at follow-up after delivery. PMID- 10389713 TI - Cost-effectiveness of estimating gestational age by ultrasonography in Down syndrome screening. AB - OBJECTIVE: To quantify the financial benefits of using ultrasound estimation of gestational age in maternal serum screening for Down syndrome. METHODS: Maternal age-specific sensitivity and false-positive rates for Down syndrome were derived for the triple test (alpha-fetoprotein, hCG, and unconjugated estriol) using gestational age based on ultrasound dating and also time from the last menstrual period (LMP). These rates were entered into a formula to determine the societal financial net benefit of Down syndrome screening. The average per-case net benefits of ultrasound- and LMP-dated pregnancies were then compared. Average net benefits were also calculated separately with ultrasound versus LMP dating for triple tests referred to our laboratory, and the additional costs associated with any post-test ultrasound scans, repeat testing, or recalculations were estimated. RESULTS: The use of ultrasound dating resulted in higher detection rates for Down syndrome and lower false-positive rates, which translated into an average per case savings to society of $33.54. For women referred to our program with LMP dating, there was an average reduction of $31.60 in net benefits, plus added costs of $14.39 attributable to extra ultrasound, repeat testing, and recalculation. CONCLUSION: When ultrasound dating is available before serum screening, it should be used preferentially to establish Down syndrome risk. Routine first-trimester ultrasound examination can be justified for women with a known LMP if the cost of the ultrasound examination is less than $46. PMID- 10389714 TI - Rapid testing and zidovudine treatment to prevent vertical transmission of human immunodeficiency virus in unregistered parturients: a cost-effectiveness analysis. AB - OBJECTIVE: To assess the potential effectiveness and costs of a program to prevent vertical transmission of human immunodeficiency virus (HIV) in parturients without prenatal care. METHODS: A decision-analysis model was constructed to compare three management strategies for unregistered women presenting in labor: 1) the current standard of treating no one; 2) HIV testing with a rapid antibody assay, followed by zidovudine treatment according to AIDS Clinical Trial Group Protocol 076 if seropositive; and 3) treating all women without rapid testing. Cost and probability data were obtained from a literature review and local estimates. Sensitivity analyses were performed for pertinent uncertainties. RESULTS: Under baseline assumptions (5% HIV prevalence, treatment efficacy of an 18% reduction in transmission rate, and lifetime cost of pediatric HIV $103,708), giving no treatment resulted in 1275 infected infants per 100,000 mother-infant pairs. The rapid-test strategy prevented 183 cases of infant HIV infection and resulted in a net savings to the medical system of $57,997 per case averted. The treat-all strategy prevented an additional 46 cases per 100,000 mother-infant pairs, but at a cost of $342,068 per additional case averted. With other estimates at baseline, rapid testing was cost-saving when the HIV prevalence exceeded 0.97%, the treatment efficacy exceeded a 5.8% reduction in the transmission rate, and the lifetime cost of pediatric HIV infection exceeded $33,625. CONCLUSION: Rapid HIV testing of unregistered parturients followed by zidovudine treatment if seropositive would be cost saving to the medical system. PMID- 10389715 TI - Selection of delivery method in pregnancies complicated by autoimmune thrombocytopenia: a decision analysis. AB - OBJECTIVE: To compare three common strategies for selecting delivery methods in term pregnancies complicated by immune thrombocytopenia by contrasting their effects on the number of severely thrombocytopenic neonates delivered vaginally and total cesarean rates. METHODS: We used decision analysis to compare three strategies to select delivery method in women with autoimmune thrombocytopenia, funipuncture at term, intrapartum fetal scalp platelet sampling with delivery mode decisions based on platelet count in the first two strategies, and no testing of fetal platelets with delivery mode determined by standard obstetric criteria. We assumed that the goal of each strategy was to minimize the number of severely thrombocytopenic neonates delivered vaginally while maintaining an acceptable cesarean rate. Severe thrombocytopenia was defined as under 50,000 platelets per microL. Probabilities with ranges (used in sensitivity analyses) were derived from the medical literature. RESULTS: Of the two testing strategies, funipuncture was clearly preferable. Funipuncture resulted in zero cases of severely thrombocytopenic neonates delivered vaginally (as did scalp sampling), with a lower overall cesarean rate compared with fetal scalp sampling (36.6% versus 69.1%). Compared with the no-testing strategy, the funipuncture strategy reduced the number of severely thrombocytopenic neonates delivered vaginally (0 versus 82 per 1000) with a modest increase in the cesarean rate (1.9 cesareans to prevent vaginal delivery of one severely thrombocytopenic neonate). CONCLUSION: Fetal scalp sampling should be abandoned in favor of funipuncture when testing for thrombocytopenia. PMID- 10389716 TI - Thoracic-fluid conductivity in peripartum women with pulmonary edema. AB - OBJECTIVE: To measure the level of thoracic-fluid conductivity associated with pulmonary edema in peripartum women by noninvasive thoracic electrical bioimpedance. METHODS: Between March 1994 and August 1996, 134 women were selected for thoracic electrical bioimpedance monitoring. Among them, 12 had pulmonary edema, 33 had severe preeclampsia, 17 had mild preeclampsia, and 72 were in uncomplicated early labor. Each subject's highest thoracic-fluid conductivity measurement was related to her clinical presentation. The Kruskal Wallis one-way analysis of variance was used to compare groups' means. A receiver operating characteristic curve was used to identify thoracic-fluid conductivity values associated with pulmonary edema. RESULTS: Pulmonary edema was associated with severe preeclampsia in ten cases, urosepsis in one case, and postoperative volume overload in one case. Other than gestational age, there were no significant differences in maternal demographics between groups. Thoracic-fluid conductivity values in women with pulmonary edema (80.6+/-18.3 kohm(-1)) were significantly higher than those in women with severe preeclampsia (62.8+/-16.3 kohm(-1)), mild preeclampsia (53.3+/-11.2 kohm(-1)), or early labor (41.3+/-6.7 kohm(-1)). Thoracic-fluid conductivity of at least 65 kohm(-1) best identified pulmonary edema (sensitivity 83.3%; specificity 86.9%; positive predictive value 38.5%; negative predictive value 98.1%). CONCLUSION: Preeclampsia was associated with increased thoracic-fluid conductivity stratified between mild and severe disease. Thoracic-fluid conductivity of at least 65 kohm(-1) was strongly associated with peripartum pulmonary edema. Women with values above 65 kohm(-1) might be candidates for medical intervention even without overt clinical symptoms. PMID- 10389717 TI - Bone mineral changes during and after lactation. AB - OBJECTIVE: To investigate variations in bone mineral density during lactation and throughout the 12 months after scheduled cessation of lactation in relation to the resumption of ovarian function. METHODS: Three hundred eight mothers who decided to lactate were scheduled to fully breast-feed for 6 months, followed by a 1-month weaning period, and then suppress lactation with cabergoline. Their bone mineral density variations were compared with those of a control group of nonlactating mothers during the first 18 months postpartum. Half the lactating women were given daily oral calcium supplements of 1 g in an open design. RESULTS: There was a significant progressive decrease in bone mineral density in lactating women over the first 6 months, followed by recovery of bone mass up to levels that at 18 months were higher than baseline. In nonlactating women, bone mineral density increased progressively after delivery, and at 18 months postpartum had increased by 1.1-1.9% compared with baseline. Compared with lactating women who resumed menstruation within 5 months of delivery, breast feeding mothers with longer amenorrhea initially lost more bone, but they also gained significantly more bone after resumption of menses, so there were no differences at 18 months postpartum. Oral calcium supplementation decreased bone loss, but had only a transient effect. CONCLUSION: A scheduled lactation period of 6 months, followed by a 1-month weaning period, allowed bone mineral density to reach higher values compared with early postpartum, regardless of calcium supplementation and duration of postpartum amenorrhea. PMID- 10389718 TI - The effect of aging on ovarian volume measurements in infertile women. AB - OBJECTIVE: To test the hypothesis that aging is associated with a decrease in ovarian volume, and that the FSH level and volume are correlated inversely. METHODS: One hundred nine women who had 73 in vitro fertilization cycles and 36 ovulation induction cycles were analyzed. Basal FSH and estradiol (E2) levels were measured on cycle day 3, and ovarian volume was measured and antral follicles were counted on the day of starting gonadotropin. RESULTS: The mean age (+/- standard deviation) was 32.6+/-4.7 years. The mean FSH was 6.9+/-2.4 IU/L. The mean ovarian volume was 6.0+/-4.7 cm3. There were no significant differences between the median volumes of the left and right ovaries in individual subjects (4.6 and 4.8 cm3, respectively; interquartile range 3.0-7.3 and 3.1-7.9; P = .79). There was a significant positive correlation between age and FSH level (R = .372, P<.001), but not between age and ovarian volume (R = .039, P = .69). A significant relation was noted between FSH and the number of follicles (H = 20.8, P<.001), but not between FSH and volume (R = .102, P = .29). There was a significant decrease in the number of follicles and a higher cycle cancellation rate in women with volume smaller than 3 cm3 compared with those with volume greater than 3 cm3. CONCLUSION: Women with small ovarian volumes, low number of antral follicles, and normal basal FSH and E2 levels may have diminished ovarian reserve. PMID- 10389719 TI - Continuous, combined hormone replacement: randomized comparison of transdermal and oral preparations. AB - OBJECTIVE: To compare two new transdermal, continuous, combined formulations and an oral regimen of hormone replacement therapy (HRT) with respect to endometrial hyperplasia, bleeding patterns, and climacteric symptoms in postmenopausal women. METHODS: This was a randomized, open, parallel-group trial during 1 year in 441 postmenopausal women who received either a 10-cm2 patch of 0.025 mg estradiol (E2) and 0.125 mg norethisterone acetate, a 20-cm2 patch of 0.05 mg E2 and 0.25 mg norethisterone acetate twice weekly, or tablets of 2 mg E2 and 1 mg norethisterone acetate once daily. The efficacy variables were frequency of endometrial hyperplasia after 1 year of treatment, number of bleeding and spotting days from the fourth to sixth treatment months, relief of climacteric symptoms, and tolerability. RESULTS: The frequency of endometrial hyperplasia was no more than 2% after 1 year of treatment in all groups. One case of simple hyperplasia was detected among the women treated with 10-cm2 patches and two among those treated with oral HRT. From the fourth to sixth treatment months, amenorrhea occurred in 73%, 47%, and 66% of the women in the 10-cm2, 20-cm2, and oral HRT groups, respectively. The 10-cm2 patches and oral treatment were associated with fewer bleeding days than were the 20-cm2 patches (P<.001). During the last 3 months of the treatment year, amenorrhea was found in 100 subjects (86%) for 10-cm2 patches, 61 (65%) for 20-cm2 patches, and in 85 (79%) for oral HRT. All treatments alleviated the climacteric symptoms to a comparable extent. CONCLUSION: In postmenopausal women, 10-cm2 patches relieved climacteric symptoms and prevented endometrial hyperplasia at least as effectively as oral HRT. Amenorrhea was induced early in a high percentage of women using 10-cm2 patches and oral HRT, and these therapies seemed to be convenient, effective, and safe for estrogen deficiency symptoms in postmenopausal women. PMID- 10389721 TI - Apoptosis and Ki-67 expression in adenomyotic lesions and in the corresponding eutopic endometrium. AB - OBJECTIVE: To examine biologic and proliferative properties of adenomyotic lesions and to determine whether adenomyotic lesions originate in the basal layer of the eutopic endometrium. METHODS: We examined eutopic and ectopic endometria from 23 patients with adenomyosis. To obtain evidence for the induction of programmed cell death, apoptotic cells were identified using a modified terminal deoxynucleotidyltransferase-biotin nick end-labeling method. To evaluate cell death repressor activity, bcl-2 gene expression was examined using immunohistochemical staining. As a proliferative marker, Ki-67 expression was also examined immunohistochemically. RESULTS: In the eutopic endometrium, apoptosis was most frequently observed in epithelial cells during mid- to late secretory phases, although it was rarely found during early proliferative through early secretory phases (P<.01). In contrast, bcl-2 gene expression inversely correlated with the appearance of apoptosis. A similar tendency was observed in stromal cells. In the ectopic endometrium of adenomyosis, endometrial dating revealed that secretory change was rare, even in the secretory phase, and that induction of apoptotic cells as well as bcl-2 gene expression showed no cyclic change. In stromal cells of the ectopic endometrium, apoptosis was more frequent than was seen in the eutopic endometrium, in all menstrual phases (P<.05). Ki-67 was constantly expressed in the glandular epithelium of the ectopic endometrium, irrespective of the menstrual phases, whereas in the secretory phase it was less expressed in the eutopic endometrium of functional and basal layers (P<.01). CONCLUSION: The induction of apoptosis seems to be regulated by hormonal changes in the eutopic endometrium and has an inverse correlation with bcl-2 gene expression. The ectopic endometrium in adenomyosis is rarely influenced by hormonal change and has different biologic and proliferative properties than events observed in the eutopic endometrium findings, which strongly suggest that the adenomyotic lesion does not originate in the basal endometrium. PMID- 10389720 TI - Prevalence of urinary incontinence and associated risk factors in postmenopausal women. Heart & Estrogen/Progestin Replacement Study (HERS) Research Group. AB - OBJECTIVE: To determine the prevalence of stress, urge, and mixed urinary incontinence and associated risk factors in postmenopausal women. METHODS: Before enrollment in a 4-year, randomized trial of combination hormone therapy to prevent coronary heart disease, 2763 participants completed questionnaires on prevalence and type of incontinence. We measured factors potentially associated with incontinence including demographics, reproductive and medical histories, height, weight, and waist-to-hip circumference ratio. We used multivariate logistic models to determine independent associations between those factors and weekly incontinence by type. RESULTS: The mean (+/- standard deviation [SD]) age of the participants was 67+/-7 years; 89% were white and 8% were black. Fifty-six percent reported weekly incontinence. In multivariate analyses, the prevalence of weekly stress incontinence was higher in white than black women (odds ratio [OR] 2.8, 95% confidence interval [CI] 1.6, 5.1), in women with higher body-mass index (BMI) (OR 1.1 per 5 units, 95% CI 1.0, 1.3), and higher waist-to-hip ratio (OR 1.2 per 0.1 unit, 95% CI 1.0, 1.4). The prevalence of weekly urge incontinence was higher in older women (OR 1.2 per 5 years, 95% CI 1.1, 1.3), diabetic women (OR 1.5, 95% CI 1.1, 2.0) and women who had reported two or more urinary tract infections in the prior year (OR 2.0, 95% CI 1.1, 3.6). CONCLUSION: Stress and urge incontinence are common in postmenopausal women and have different risk factors, suggesting that approaches to risk-factor modification and prevention also might differ and should be specific to types of incontinence. PMID- 10389722 TI - Secretion of vascular endothelial growth factor in adenocarcinoma and squamous cell carcinoma of the uterine cervix. AB - OBJECTIVE: To determine whether major differences in vascular endothelial growth factor secretion exist between adenocarcinomas of the uterine cervix compared with squamous cell carcinomas. METHODS: The secretion of vascular endothelial growth factor by eight fresh cervical cancer cell preparations (four adenocarcinomas and four squamous cell carcinomas) and four established squamous cell lines was evaluated using a sensitive enzyme-linked immunosorbent assay in vitro. RESULTS: All cervical tumors secreted significant amounts of vascular endothelial growth factor, and no significant differences between fresh and established squamous cell lines were detectable. In contrast, a highly significant difference in vascular endothelial growth factor secretion was noted between fresh adenocarcinomas (mean = 2712, range between 1700 to 3500 pg/mL/10(5) cells/48 hours) when compared with fresh squamous (mean = 575, range between 200 to 950 pg/mL/10(5) cells/48 hours) or established squamous cervical carcinoma cell lines (mean = 712, range between 400 to 1000 pg/mL/10(5) cells/48 hours) (F-test, P< or =.001). CONCLUSION: These data strongly suggest that major differences in the secretion of vascular endothelial growth factor exist between squamous cell carcinoma and adenocarcinomas of the uterine cervix. Therefore, at least some of the differences in the natural biologic behavior of these two histologic types of cervical cancer, including the propensity for earlier lymphatic and hematogenous metastasis as well as the lower response to radiation treatment, could be related to major differences in the secretion of this powerful angiogenic and immunosuppressive cytokine. PMID- 10389723 TI - Entry force and intra-abdominal pressure associated with six laparoscopic trocar cannula systems: a randomized comparison. AB - OBJECTIVE: In trocar-cannula systems, increased entry force could result in loss of operator control, a potential cause of serious visceral and vascular injuries. We developed a system to measure entry force and intraperitoneal pressure to evaluate and compare trocar-cannula systems. METHODS: Six laparoscopic trocar cannula systems of similar diameter (12 mm) were tested (two pyramidal, two cutting-dilating, and two blunt conical) using a white swine model. All six systems were inserted into each of 12 subjects with location designated by random allotment (72 insertions). During each insertion, intraperitoneal pressure and entry force were measured using a system consisting of a gas-gas transducer, a 50 lb load cell, and a multichannel data acquisition board. Mean entry force and intraperitoneal pressure were compared using mixed-model analysis of variance. RESULTS: Mean entry force measurements were as follows: pyramidal 9.01 lb and 13.48 lb, cutting-dilating 9.94 lb and 16.46 lb, and blunt conical 19.15 lb and 31.91 lb. Intraperitoneal pressure changes generally reflected measured entry force. CONCLUSION: The system successfully measured both entry force and resultant intraperitoneal pressure. Pyramidal trocar-cannula systems required the lowest force for entry. These differences in entry force have potential clinical implications related to the risk of visceral and vascular injury. Intraperitoneal pressure measurement could be used as a surrogate for insertional force measurement. PMID- 10389724 TI - Incision characteristics associated with six laparoscopic trocar-cannula systems: a randomized, observer-blinded comparison. AB - OBJECTIVE: Laparoscopic trocar-cannula systems of different design but similar internal diameter result in incisions of varying dimensions. Such variations might affect the incidence of incisional complications, such as dehiscence and hernia. We developed a system to measure associated fascial defects and then used the techniques to compare the defects resulting from different trocar-cannula systems. METHODS: This was a randomized, observer-blinded study. Six laparoscopic trocar-cannula systems of similar diameter (12 mm) were tested (two pyramidal, two blunt conical, and two cutting-dilating) using a white swine model. All systems were inserted into each of 12 subjects, with location designated by random allotment (total 72 insertions). The fascial defects were exposed and then directly measured for incisional length and area by an observer blinded to the system used. Means of each outcome variable (incisional length and area) were compared using factorial analysis of variance. RESULTS: The values for mean incisional areas were as follows: cutting-dilating 28.73 mm2 and 31.09 mm2, pyramidal 18.25 mm2 and 26.75 mm2, and blunt conical 10.00 mm2 and 12.33 mm2. Mean maximal incisional lengths were similar among all trocar-cannula systems. CONCLUSION: Blunt conical trocar-cannula systems resulted in significantly smaller fascial defects compared with the widely used pyramidal and the two cutting-dilating trocar-cannula systems tested. These differences have potential clinical implications. For example, smaller fascial defects could reduce risk of incisional hernia and dehiscence. PMID- 10389725 TI - Major complications of laparoscopy: a follow-up Finnish study. AB - OBJECTIVE: To examine recent figures on major laparoscopic complications in Finland. METHODS: This was a nationwide record-linkage study from January 1995 through December 1996 including all Finnish hospitals performing gynecologic laparoscopies. Data files of the National Patient Insurance Association and the Finnish Hospital Discharge Register were used. Data were compared with previous results from 1990 to 1994. RESULTS: Among 32,205 gynecologic laparoscopies, 130 major complications were noted. The total complication rate was 4.0 per 1000 procedures: 0.6 per 1000 in diagnostic laparoscopies, 0.5 per 1000 in sterilization, and 12.6 per 1000 in operative laparoscopies. Intestinal injuries were reported in 0.7 per 1000, incisional hernias in 0.3 per 1000, urinary tract injuries in 2.5 per 1000, major vascular injuries in 0.1 per 1000, and other injuries in 0.5 per 1000 gynecologic laparoscopic procedures. Seventy-five percent (88 of 118) of the major complications in operative laparoscopies occurred during hysterectomies. The total major complication rate decreased from 4.9% in 1993 to 2.3% in 1996 (chi2 = 8.55, P = .003), but the incidence of ureteral injuries remained stable, at about 1% of laparoscopic hysterectomies. Ureteral injuries were most common in local hospitals (2.6%), followed by central (1.1%) and university hospitals (0.9%). From 1990 through 1996, the relative risk for ureteral injury in laparoscopic hysterectomies, compared with other operative laparoscopies was 29.0 (95% confidence interval [CI] 13.3, 63.0), for bladder injury 13.0 (95% CI 6.0, 28.2), for intestinal injury 1.3 (95% CI 0.6, 2.5), and for major vascular injury 0.4 (95% CI 0.1, 3.6). Compared with the figures for 1990-1994, all major complications in operative laparoscopies increased, from 0 per 1000 in 1990 to 14.0 per 1000 in 1996 (chi2 = 20.28, P<.001), but part of this increase was due to the increased proportion of laparoscopic hysterectomies. CONCLUSION: Laparoscopic hysterectomies are still associated with a stable 1% risk of ureteral injury, whereas other major complications were decreasing until 1996. Complications in other laparoscopic procedures generally are rare. PMID- 10389726 TI - Test-retest reliability of the cough stress test in the evaluation of urinary incontinence. AB - OBJECTIVE: To determine the test-retest reliability of the cough stress test in women who present with complaints of urinary incontinence. METHODS: This prospective observational study involved 50 women between the ages of 28 and 78 years with primary complaints of urinary incontinence. All subjects underwent a pelvic examination, cystometrogram, and a cough stress test. The cough stress test was performed in the standing position at a bladder volume of 300 mL or at maximum cystometric capacity if it was less than 300 mL. The results of the cough stress test were recorded as positive if urine loss occurred with a cough or as negative if no urine loss was seen and the bladder volume was recorded. The women returned in 1-4 weeks for a second cough stress test performed at the same bladder volume as at the initial examination. RESULTS: Of the 50 women studied, 45 (90%) had similar results with both cough stress tests. Thirty-five had a positive cough stress test on the initial examination, and 32 of these patients (91%) also had a positive cough stress test at a repeat visit. Fifteen patients had a negative initial cough stress test, and 13 of these 15 (87%) had a negative cough stress test on repeat examination. All 20 patients diagnosed as having pure genuine stress incontinence had a positive cough stress test on initial and repeat examinations. Of the 15 patients diagnosed with detrusor instability or sensory urge incontinence, 13 (86%) had negative cough stress tests on initial and repeat examinations. Of the 15 patients diagnosed with mixed incontinence, 12 (80%) had positive cough stress tests on initial and repeat examinations. CONCLUSION: The cough stress test appears to be a reliable test. The reliability is more consistent in women with a diagnosis of pure genuine stress incontinence. PMID- 10389727 TI - Once-daily valacyclovir hydrochloride for suppression of recurrent genital herpes. AB - OBJECTIVE: To document the frequency of genital herpes recurrences in men and women with histories of recurrent genital herpes during 1 year of continuous, suppressive therapy with valacyclovir hydrochloride (HCl). METHODS: In an open label clinical trial conducted at 11 centers, 127 subjects (46 women and 81 men) with histories of recurrent genital herpes (at least 6 recurrences per year) were treated with valacyclovir HCl (500 mg once daily), and their clinical status was followed up for 1 year. Genital herpes recurrences were documented in diaries, and quarterly clinic visits were made for evaluating lesion recurrences and drug safety. In cases of recurrence, subjects self-treated with valacyclovir HCl 500 mg twice daily for 5 days, then resumed once-daily treatment. RESULTS: After the first 3 months of suppressive therapy, 81% of subjects were free of recurrence. Recurrence-free rates remained undiminished during the second, third, and fourth quarters (84%, 84%, and 91%, respectively) and were similar for men and women. Thirty of 46 women (65%) and 56 of 81 men (69%) remained recurrence free during the study and therapy was well tolerated. Adverse events were mild, infrequent, and not considered related to the study drug. CONCLUSION: Valacyclovir HCl was highly effective and well tolerated as continuous suppressive therapy in men and women with recurrent genital herpes. Potential benefits of the once-daily regimen of valacyclovir HCl include improved patient compliance. PMID- 10389728 TI - Urine free beta-hCG and total estriol for Down syndrome screening during the second trimester in an Asian population. AB - OBJECTIVE: To evaluate second-trimester free beta-hCG and total estriol (E3) in the maternal urine as markers for Down syndrome screening in an Asian population. METHODS: Free beta-hCG and total E3 were measured in the urine samples of 28 Taiwanese Down syndrome pregnancies and 268 unaffected singleton pregnancies at 14-25 weeks. Results were normalized to urine creatinine concentrations and converted to multiples of the median (MoM) levels. Gestational ages were estimated by ultrasound measurements. RESULTS: Median values of free beta-hCG, total E3, free beta-hCG to total E3 ratio, and the free beta-hCG to total E3 MoM ratio in Down syndrome pregnancies were 4.75 MoM, 0.66 MoM, 8.99 MoM, and 9.51, respectively. At a 5% false-positive rate, the observed detection rates were 36% (ten of 28) with total E3, 71% (20 of 28) with free beta-hCG, 68% (19 of 28) with free beta-hCG/total E3, and 71% (20 of 28) with free beta-hCG/total E3 MoM. When combined with maternal age, the expected detection rates were 65% with total E3, 71% with free beta-hCG, 76% with free beta-hCG/total E3, 80% with free beta hCG/total E3 MoM, and 89% when combining free beta-hCG, total E3, and maternal age. CONCLUSION: Urine free beta-hCG and total E3 are useful markers for Down syndrome screening during the second trimester in Taiwanese women. PMID- 10389729 TI - Modern obstetric management and outcome of infants with gastroschisis. AB - OBJECTIVE: To determine whether outcomes of infants with gastroschisis differed by mode or site of delivery, diagnostic method, or when maternal-fetal medicine consultation was given. METHODS: Charts of 32 infants born at the University of Mississippi Medical Center or admitted to the neonatal intensive care unit between September 1992 and June 1998 were reviewed for maternal demographic characteristics and neonatal outcomes. Statistical analysis was done using Student t test, analysis of variance, chi2, and Kruskal-Wallis test with P<.05 considered statistically significant. RESULTS: There were no statistically significant differences in neonatal outcomes by method or site of delivery, diagnostic method, or maternal-fetal medicine consultation before delivery. Infants delivered vaginally had higher Apgar scores at 1 and 5 minutes (9 versus 7 and 9 versus 8, respectively, P<.05). Vaginally delivered infants required more days of antibiotic therapy than those delivered abdominally (10 versus 3 days, P<.05) but had a shorter interval to enteral feedings (15 versus 30 days, P<.05). CONCLUSION: Outcomes of infants with isolated gastroschisis were not significantly affected by method or site of delivery, diagnostic method, or maternal-fetal surveillance. Although the findings of this investigation were largely negative and the statistical power limited due to the rarity of this fetal disruption, small series of cases of gastroschisis need to be analyzed to resolve current controversies surrounding optimal treatment of gastroschisis. PMID- 10389730 TI - Change in cervical length after prophylactic cerclage. AB - OBJECTIVE: To determine changes in length of incompetent cervices after cerclage, using transvaginal ultrasound. METHODS: Patients were enrolled in a prospective, observational study under an Institutional Review Board-approved protocol. McDonald or Shirodkar sutures were placed according to physician preference. Pre- and postcerclage cervical lengths were measured within 72 hours of the procedure. At each examination, the first measurement was discarded, and a mean of the subsequent three measurements was calculated. RESULTS: Twenty-one Shirodkar and ten McDonald operations were done. The mean (+/- standard deviation) precerclage cervical length was 2.7+/-0.9 cm and the postcerclage cervical length was 3.6+/ 0.9 cm (P<.001, paired t test). CONCLUSION: Prophylactic cerclage results in measurable increases in cervical length, which might contribute to the success of the procedure. Further study is needed to determine whether the degree of cervical lengthening after cerclage predicts term delivery. PMID- 10389731 TI - Neonatal morbidity at 34-37 weeks: the role of ruptured membranes. AB - OBJECTIVE: Evaluate neonatal morbidity in deliveries occurring between 34 0/7 and 36 6/7 weeks' gestation, comparing outcomes in pregnancies complicated by preterm premature rupture of membranes with those in which delivery occurred with intact membranes prior to the onset of labor. METHODS: The obstetric database was reviewed for a 5-year period. Healthy gravidas delivering nonanomalous singleton gestations from vertex presentations were evaluated, with corticosteroid or antibiotic administration or both noted. The neonatal database was reviewed for the following complications: admission to the neonatal intensive care unit, need for assisted ventilation, and development of hyaline membrane disease, bronchopulmonary dysplasia, intraventricular hemorrhage, necrotizing enterocolitis, or culture-proven sepsis. Groups were compared using chi2 tests. The power of this study to detect a ten-fold decrease in the likelihood of neonatal complications at the P<.05 significance level was greater than 90%. RESULTS: Of 853 eligible pregnancies, 414 (48.5%) gravidas had ruptured membranes prior to the onset of active labor. No difference existed between groups in the number of patients who had received corticosteroids during pregnancy, but patients with ruptured membranes were more likely to have received antibiotics prior to delivery. No neonatal deaths occurred, and neonatal morbidity was low in both groups. CONCLUSION: No clinically significant difference exists in neonatal outcome between 34 0/7 and 36 6/7 weeks' gestation as the result of membrane status prior to the onset of labor. PMID- 10389732 TI - Natural protective mechanisms against endoscopic white-light injury in the fetal lamb eye. AB - OBJECTIVE: To evaluate light transmission, possible light trauma, and techniques for protection of the fetal eyes during intrauterine videoendoscopic surgery in a sheep model. METHODS: In vitro studies were done at various gestational ages, including spectrometry of light output by a halogen light source and telescope and light transmission by spectrophotometry in the range of 180-3000 nm through amniotic fluid (AF) and fetal eyelids. In vivo electron-microscopic, morphologic analysis of the retinas of 65-, 95-, 108-, and 112-day-old fetal lambs with (n = 8) and without (n = 8) 30 minutes' light exposure to the open eye was also done. RESULTS: The light spectrum at the tip of the telescope was 400-750 nm, with a maximum irradiance of 3x10(-3) W/cm2 at 580 nm. In the ultraviolet spectrum (less than 300 nm), irradiance was less than 0.5x10(-3) W/cm2. Light transmission through ovine AF ranged from 30% at 300 nm to 89% at 700 nm at 84 days' gestation, decreasing to less than 0.01% (300 nm) and 70% (700 nm) at 112 days. Fetal eyelids did not transmit more than 1% of light (any wavelength). After direct in vivo light exposure, no retinal damage was found. Photoreceptors were present from 108 days onward, but chromophores were scant or absent at all ages studied. CONCLUSION: The light spectrum of a standard endoscope is limited to 400 750 nm; ultraviolet light is filtered out. The AF transmits harmful blue light poorly. Fetal eyelids seem to protect the eye by extremely low transmission and light dispersion. Even with the eye open, no morphologic retinal damage was found. The strong light sources used with fetal endoscopy did not appear to pose a threat to the fetal retina. PMID- 10389733 TI - Matrilineal transmission of birth weight in the rhesus monkey (Macaca mulatta) across several generations. AB - OBJECTIVE: To investigate how secular trends in maternal weight characteristics, in response to living in a permissive laboratory environment, influence intergenerational trends in birth weight in the rhesus monkey (Macaca mulatta) and to assess the role of female offspring in perpetuating these matrilineal traits. METHODS: A multigenerational data set was used to evaluate the relationship between familial and contemporaneous pregnancy factors and infant birth weight across several generations. These records provided 25 years of information on the maternal and paternal ancestries and reproductive histories, gestation lengths, and birth weights for 1321 infants. RESULTS: Pregnancy weight gain, gestation length, and maternal familial factors were the most important predictors of infant birth weight, followed by infant sex, paternity, and maternal pregravid weight (P<.001 for each variable). Furthermore, the trend in fetal growth across generations followed a matrilineal pattern of transmission that was much more pronounced for female than male offspring (P<.001). Although secular increases in maternal pregravid weight and pregnancy weight gain were detected, the upward shift in female birth weight was not explained solely by these changes in maternal weight parameters. CONCLUSION: With the delivery of ample nutrition and health care in a laboratory setting, there was a dramatic increase in the birth weight of daughters within certain matrilines, providing evidence that an intrauterine mechanism transmitted through female progeny can regulate fetal development. Further, the upward trend in female birth weight had a beneficial influence on the reproductive performance of female descendants in those lineages. PMID- 10389734 TI - Cystic fibrosis and chromosome abnormalities associated with echogenic fetal bowel. AB - OBJECTIVE: To determine the prevalence of cystic fibrosis mutations and chromosome abnormalities in the fetuses of a heterogeneous population of pregnant women referred for prenatal testing for echogenic fetal bowel. METHODS: Fetal or parental samples obtained after a second-trimester sonographic finding of echogenic fetal bowel were submitted to a referral diagnostic laboratory during a 2-year period. Results of DNA testing and karyotyping on these samples were analyzed to determine the prevalence of cystic fibrosis transmembrane reductase gene mutations and chromosome abnormalities. RESULTS: Of 244 cases tested, two fetuses were positive for two cystic fibrosis mutations. This rate (0.8% or two of 244) is 20 times higher than the general white population rate of one per 2500. In a third case, both parents were carriers but the fetus was not tested. Nine (8%) of 113 fetuses tested had one cystic fibrosis mutation. Of 106 fetuses for whom chromosome results were available, three (2.8%) fetuses had a chromosomal abnormality: two had trisomy 21 and one had Klinefelter syndrome. A fourth fetus carried a de novo, apparently balanced, 5;12 translocation. CONCLUSION: These laboratory results are representative of a broad spectrum of clinical settings and indicate a generalized increased risk associated with this sonographic finding. Therefore, when a second-trimester sonographic diagnosis of fetal echogenic bowel is made, fetal testing for both cystic fibrosis and chromosome abnormalities is warranted. PMID- 10389735 TI - Effect of causing fetal cardiac asystole on second-trimester abortion. AB - OBJECTIVE: To compare second-trimester abortions with prostaglandin (PG) E2, with and without pretreatment-induced fetal death. METHODS: A retrospective chart review of all vaginal PG E2-induced abortions at Westchester Medical Center between January 1996 and April 1998 was done. Only women who sought terminations between 18 and 24 weeks' gestation by prostaglandin induction were included. These abortions were predominantly secondary to fetal structural and chromosomal anomalies. The study population was subdivided into groups based on the use of pretreatment cardiac puncture with potassium chloride. The groups were compared for maternal, fetal, and procedural characteristics. The chi2 test, Student t test, and Wilcoxon rank-sum test were used for analysis. RESULTS: There were no differences between the cardiac puncture and control groups when compared for various maternal and procedural characteristics, fetal weight, and the need for curettage for retained products of conception. However, the required median doses of PG E2 and the initiation to expulsion interval were significantly lower in the cardiac puncture group compared with the control group (2.0 doses compared with 3.0 doses, P<.001; 570 minutes compared with 890 minutes, P = .006). CONCLUSION: Pretreatment-induced fetal death significantly reduced the interval to expulsion and doses of PG E2 required for late second-trimester abortion. PMID- 10389736 TI - Informed consent, patient choice, and physician responsibility. PMID- 10389737 TI - Effect of residency program merger on undergraduate medical student education in obstetrics and gynecology. AB - OBJECTIVE: To evaluate the effect of residency program merger on third-year medical student clerkships using student evaluations of their experiences and standardized subject examination scores. METHODS: End-of-clerkship ratings from student evaluations and National Board of Medical Examiners standardized subject examination scores in obstetrics and gynecology were used from clerkship sites where three separate military residency programs in obstetrics and gynecology recently merged into two new programs. Mean student evaluation scores and subject examination scores for the year preceding and the year following the merger were compared. RESULTS: The mean differences in medical student evaluation scores before and after merger of the residency programs were 0.1 (Mann-Whitney rank sum, P = .1), -0.1 (Mann-Whitney rank sum, P = .8), and 0.2 (Mann-Whitney rank sum, P = .3). The mean differences for subject examination scores before and after merger of the residency programs were -3.5 (95% confidence interval [CI] 33.2, 26.2; paired t test), -30.1 (95% CI -58.9, -1.4; paired t test), and -35.3 (95% CI -74.8, 4.3; paired t test). CONCLUSION: Merger of residency programs in obstetrics and gynecology does not appear to have a deleterious effect on medical students' satisfaction with the clerkship or their performance on standardized subject examinations at our institution. PMID- 10389738 TI - Angiogenesis of the endometrium. AB - OBJECTIVE: To present current data pertaining to angiogenesis of the endometrium throughout the normal menstrual cycle and in benign and neoplastic diseases of the endometrium. SOURCES: We conducted a computerized search of MEDLINE, Current Contents, and Index Medicus for relevant studies in the English literature published between January 1966 and October 1998. STUDY SELECTION: We reviewed all studies that included human and animal models of angiogenesis of normal cyclic endometrium and benign and neoplastic endometrial diseases. TABULATION, INTEGRATION, AND RESULTS: Angiogenesis is important to cyclic, regenerating endometria and disease processes including dysfunctional uterine bleeding, response to exogenous hormonal treatment, bleeding associated with intrauterine contraceptive devices, uterine leiomyomata, endometriosis, complex endometrial hyperplasia, and endometrial carcinoma. CONCLUSION: In the future, knowledge of specific angiogenic patterns of various disease processes might improve application of antiangiogenic medications in therapies for benign and neoplastic diseases of the endometrium. PMID- 10389739 TI - Serious group A streptococcal infection around delivery. AB - OBJECTIVE: To differentiate the features of serious, perinatal, group A streptococcal infection from other types of streptococcal toxic shock syndrome. DATA SOURCES: Thirty-eight obstetric cases that were fatal or fulfilled the criteria of Centers of Disease Control and Prevention for streptococcal toxic shock syndrome were reviewed. Three cases were from Asahi General Hospital, 26 from MEDLINE (1966-December 1998), four from Japana Centra Revuo Medicina (1987 November 1998) using the search terms "Streptococcus," "Streptococcus pyogenes," "Streptococcal infection," "pregnancy," "labor," "delivery," "sepsis," and "shock," and five from official records of the Ministry of Health and Welfare of Japan. Cases of early pregnancy or cases that had unclear intervals between delivery and deterioration were eliminated. INTEGRATION AND RESULTS: The 30 cases were divided into two groups by interval between delivery and deterioration. Seventeen cases deteriorated before, during, or within 12 hours of delivery (perinatal group). They were compared with 13 cases of the puerperal group. The mortality rates for infants and mothers in the perinatal group were higher than those of the puerperal group (infant: ten of 17 versus zero of 13, mother: 15 of 17 versus seven of 13). The other differences of description were unusually strong labor (eight of 17 versus one of 13), obvious serious inflammation (zero of 17 versus ten of 13), and evidence of serious sepsis (eight of 17 versus zero of 13). Purulent myometritis without neighboring inflammation was found in our three cases. In ten cases, subjects or their family members had preceding sore throats. CONCLUSION: The cases in the perinatal group had characteristic features. We suspected that after invading the myometrium through the upper respiratory tract, large amounts of cocci were dispersed into the systemic circulation of the mother by active uterine contractions caused by purulent myometritis. Unusual clinical signs were important for diagnosis. PMID- 10389740 TI - Station and cervical dilation at epidural placement in predicting cesarean risk. PMID- 10389741 TI - The Zavanelli maneuver: 12 years of recorded experience. PMID- 10389742 TI - Cytology alone versus cytology and cervicography for cervical cancer screening: a randomized study. PMID- 10389743 TI - Perineal talc exposure and subsequent epithelial ovarian cancer: a case-control study. PMID- 10389744 TI - Station at onset of active phase in nulliparous patients and risk of cesarean delivery. PMID- 10389746 TI - A case-control study of childhood leukemia in southern Ontario, Canada, and exposure to magnetic fields in residences. AB - A population-based case-control study was conducted in Ontario, Canada, to assess the relation between the risk of childhood leukemia and residential exposure to magnetic fields. Participating subjects consisted of 201 cases, diagnosed at 0 to 14 years of age during 1985-1993, ascertained from the records at the Hospital for Sick Children (Toronto), and 406 individually matched controls. Where possible, point-in-time measurements of magnetic fields were made in all residences occupied by subjects during the period of inquiry in the defined catchment area. Three different classification schemes of wire code were assigned to each residence. Detailed information was collected by interviewer-administered questionnaires, which enabled risk estimates to be adjusted for socio-economic characteristics, medical history of parent(s) and child and environmental exposures. Inconsistent elevations in risk were associated with time-weighted averages of magnetic fields both inside and outside the home for subjects having residential point-in-time measurements that represented at least 70% of their etiological period. These risks increased in magnitude when analysis was restricted to children under 6 years of age at diagnosis or to those with acute lymphoblastic leukemia. For children younger than 6 years at diagnosis, outside perimeter measurements of the residence, > or = 0.15 microT, were associated with increased leukemia risk (OR = 3.45, 95% CI = 1.14-10.45). Evaluation of different exposure times for point-in-time magnetic field measurements and wire configuration suggested that exposures earliest in the etiological period were associated with greater risks for children diagnosed at a younger age (OR = 2.50, 95% CI = 1.14-5.49). Our findings did not support an association between leukemia and proximity to power lines with high current configuration. PMID- 10389745 TI - Role of medical history in brain tumour development. Results from the international adult brain tumour study. AB - In an international population-based case-control study carried out in 8 centres in 6 countries, we investigated the role of specific medical conditions in the aetiology of brain tumours in adults. Recruited were 1,178 glioma and 331 meningioma cases and 2,493 age- and gender-matched population controls. Only medical conditions occurring at least 2 years before brain tumour diagnosis were considered. Relative risks (RRs) and 95% confidence intervals (CIs) were estimated using a conditional logistic regression model. Heterogeneity between centres was tested. No association between meningioma and previous medical conditions was observed. For glioma, there was an increased risk associated with epilepsy (RR = 6.55, 95% CI 3.40-12.63), but this was considerably weaker for epilepsy of more than 20 years duration. The risk remained elevated after adjustment for use of anti-epileptic drugs. There was a statistically significant inverse association between glioma and all allergic diseases combined (RR = 0.59, 95% CI 0.49-0.71); this was also observed for specific allergic conditions, namely, asthma and eczema. Subjects who reported a history of infectious diseases (e.g., colds, flu) showed a 30% reduction in risk (RR = 0.72, 95% CI 0.61-0.85). The decreased risks for glioma in subjects reporting a history of allergic conditions or infectious diseases may indicate an influence of immunological factors on the development of glioma. The association between glioma and epilepsy has to be interpreted cautiously and needs further investigation. PMID- 10389747 TI - Risk factors for adenocarcinoma of the small intestine. AB - We have investigated the relation between alcohol, tobacco and dietary habits and risk of adenocarcinoma of the small intestine using data from 2 hospital-based case-control studies on intestinal cancers conducted in 6 Italian centres between 1985 and 1996. Cases were 23 patients below age 75 years with adenocarcinoma of the small intestine. Controls were 230 patients admitted to hospital for a wide spectrum of acute, non-neoplastic, non-digestive tract diseases, matched to cases on sex, age, study and centre. Odds ratios (ORs) were estimated using conditional logistic regression. Alcohol and tobacco consumption did not increase the risk of adenocarcinoma of the small intestine. The risk appeared to be directly related to intake of bread, pasta or rice (OR = 3.8), sugar (OR = 2.9) and red meat (OR = 4.6), and inversely to coffee (OR = 0.4), fish (OR = 0.3), vegetables (OR = 0.3) and fruit (OR = 0.6). Our results suggest that dietary correlates of adenocarcinoma of the small intestine are similar to those of colon cancer and at least of the same magnitude. While the present data are inconsistent with a major effect of tobacco or alcohol, a moderate association between these factors and small bowel cancer may have been obscured by the play of chance. PMID- 10389748 TI - Association between N-acetyltransferase 2 (NAT2) genetic polymorphism and development of breast cancer in post-menopausal Chinese women in Taiwan, an area of great increase in breast cancer incidence. AB - The incidence of breast cancer has increased greatly in Taiwan over the past 2 decades. Increased exposure to environmental carcinogens, including aryl aromatic amines, as a result of the economic boom, is suspected to be one factor contributing to this increase. The enzyme N-acetyltransferase 2 (NAT2) determines the rate of metabolism of aryl aromatic amines, and therefore the NAT2 slow acetylator genotype is associated with an increased risk of cancer. Our present case-control study of 150 breast cancer patients and 150 healthy controls in Taiwan was performed to explore the association between NAT2 genetic polymorphism and individual susceptibility to breast cancer. A structured questionnaire was used to collect relevant information regarding all known or suspected risk factors of breast cancer. The NAT2 genotype was determined using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay in 139 cases and 133 controls, and 28.8% and 21.1%, respectively, were found to have slow acetylator genotypes. Multivariate analysis, simultaneously considering other risk factors, including age at menarche, nulliparity or age at first full term pregnancy, body mass index (BMI), hormone replacement therapy (HRT) and smoking status, showed that the NAT2 slow acetylator genotype was associated with an increased risk with borderline significance (Odds Ratio, 1.81; 95% CI, 1.01 3.31). Interestingly, this association was not significant in premenopausal women, but was significant in post-menopausal women. Further stratification of our study subjects based on different risk factor status showed that the increased risk for an NAT2 slow acetylator was more marked in post-menopausal women who were not using HRT or who had a lower BMI. Our findings suggest that NAT2 polymorphism is a susceptibility factor for breast cancer in Taiwanese women, and that NAT2-metabolized carcinogens are probably present in the environment and may be associated with induction of breast cancer. PMID- 10389749 TI - Multiple primary tumors of the upper aerodigestive tract: is there a role for constitutional mutations in the p53 gene? AB - Head-and-neck cancer (HNC) patients have a high risk of developing second primary tumors of the upper aerodigestive tract, the main cause of death. Although the roles of tobacco and diet in multiple head-and-neck carcinogenesis have been thoroughly investigated, little is known about individual genetic susceptibility factors involved in this process. Genomic instability, reflecting the propensity and the susceptibility of the genome to acquire multiple alterations, could be considered a driving force behind multiple carcinogenesis. Mutation of the p53 tumor-suppressor gene has been proposed to play an important role in this process. Therefore, we evaluated the incidence of inherited p53 germ-line alteration(s) in a population of 24 consecutive HNC patients and their first degree relatives affected by multiple malignancies as well as the occurrence of p53 somatic acquired mutation(s) in 16 cancers, including first and second primaries from 5 HNCs of the same group. Mutations in exons 4-11 of the p53 gene were investigated using SSCP-PCR analysis and DNA sequencing. Analysis was extended to the peripheral blood and cancer biopsies available from first-degree relatives of cancer-prone families with p53 germ-line mutations. p53 germ-line mutations were identified in the peripheral blood and corresponding cancers of 3 HNC patients who had multiple malignancies. The only missense mutation detected was mapped in exon 6; it is a GTG to GAG substitution with an amino acid change from Val to Glu at codon 197. The remaining 2 p53 germ-line mutations were single nucleotide substitutions without amino acid change in exon 6 (codon 213, CGA to CGG) and in exon 8 (codon 295, CCT to CCC), respectively. These mutations were found in HNC patients with a family history of cancer. Abnormal expression of wild-type p53 protein in normal and pathological tissues from patients with the same sense single-nucleotide substitutions was detected by immuno-histochemistry. PMID- 10389750 TI - Different frequencies of p53 codon-249 hot-spot mutations in hepatocellular carcinomas in Jiang-su province of China. AB - Environmental carcinogens often induce specific mutations in the p53 gene, apparent in tumors. The relation between aflatoxin B1 (AFB1 )-related hepatocellular carcinomas (HCCs) and hot spot at codon 249 of the p53 gene has received a great deal of attention, but its significance is still controversial. To clarify this problem, we analyzed the p53-mutational status of HCCs in Jiang su province in China, where AFB1 contamination of the staple food significantly differs between the northern and southern parts (prominent only in the latter), while other conditions are quite similar. Background liver status and mutations in exons 5 to 8 of p53 in a total of 31 cases were divided approximately equally between the 2 areas. In all, 15 tumors exhibited a total of 17 mutations in the p53 gene; 9 cases from the southern part of the province had the hot-spot mutation at codon 249 (9/16, 56%), but only one case from the northern part (1/15, 8%). These results suggest that AFB1 contamination may correlate with codon-249 mutations in HCC. PMID- 10389751 TI - A cohort study of oral contraceptive use and risk of benign breast disease. AB - The purpose of the cohort study reported here was to investigate the association between oral contraceptive use and risk of benign breast disease (BBD), overall and by histological subtype, within the 56,537 women in the Canadian National Breast Screening Study (NBSS) who completed self-administered lifestyle and dietary questionnaires. The NBSS is a randomized controlled trial of screening for breast cancer in women aged 40-59 at recruitment. Cases were the 2,116 women in the dietary cohort who were diagnosed with biopsy-confirmed incident BBD. For comparative purposes, a subcohort consisting of a random sample of 5,681 women (including 197 subjects with incident BBD) was selected from the full dietary cohort. After exclusions for various reasons, the analyses were based on 2,116 cases and 5,338 non-cases. There was an inverse association between use of oral contraceptives and risk of all types of BBD combined. The reduction in risk was confined largely to proliferative forms of BBD (BPED), and in particular, to those forms of BPED without histological atypia, in whom there was a progressive reduction in risk with increasing duration of use (the IRR (95% CI) for use of more than 7 years was 0.64 (0.47-0.87)); risk of BPED with atypia was increased somewhat in association with oral contraceptive use (the IRR (95% CI) for use of more than 7 years was 1.43 (0.68-3.01 )), but not in a dose-dependent manner. The results were similar when examined separately in the screened and control arms of the NBSS and for screen-detected and interval-detected BPED. PMID- 10389752 TI - Expression of Smad proteins in human colorectal cancer. AB - Escape from transforming growth factor-beta (TGF-beta)-induced inhibition of proliferation has been observed in many tumor cells and may contribute to loss of growth control. Smad proteins have been identified as major components in the intracellular signaling of TGF-beta family members. In this study, we examined the expression of receptor-activated, common-mediator and inhibitory Smads by immunohistochemistry in human colorectal cancers. We found increased expression of receptor-activated Smads in a fraction of the tumor cells, while no immunostaining for Smad2, Smad3 or Smad5 and only occasional staining for Smad1/8 was found in epithelial mucosa of normal colon. No or only weak staining for receptor-activated Smads, common-mediator Smad4 and inhibitory Smads was observed in the tumor stroma. Common-mediator Smad4 and inhibitory Smads were detected in cells of both tumor and normal tissues. We observed a distinct pattern of Smad4 immunostaining of epithelial cells along colon crypts, with high expression in zones of terminal differentiation. Our data show selective up-regulation of receptor-activated Smad proteins in human colorectal cancers and suggest involvement of Smad4 in differentiation and apoptosis of surface epithelial cells of normal crypts. PMID- 10389753 TI - Uniform distribution of HPV 16 E6 and E7 variants in patients with normal histology, cervical intra-epithelial neoplasia and cervical cancer. AB - High-risk human papillomaviruses (HPV), particularly HPV 16, are associated with invasive cervical cancer (ICC), and persistent high-risk HPV infection is considered to be a marker for progressive cervical intra-epithelial neoplasia (CIN). However, most high-risk, HPV-infected, pre-cancerous lesions will not progress to invasion. Several reports suggest that specific HPV 16 E6 and/or E7 sequence variations may be associated with a high risk for progression. No data from German patients have so far been reported. Therefore, we analyzed intra-type variations of these oncogenes in women with normal histology or CIN 1 (< or = CIN 1), CIN 2/3 or ICC. Cervical scrapes from 75 patients with normal histology or CIN and biopsies from 37 ICC patients all positive for HPV 16 were analyzed. The open reading frames of oncogenes HPV 16 E6 and E7 were amplified by nested PCR followed by primer cycle sequencing. From each cervical scrape, 2 independent PCR amplicons were generated and sequenced from both orientations. The prototype sequence of HPV 16 E6 and E7 was identified in 33% and 87% of < or = CIN 1, in 62% and 69% of CIN 2/3 and in 43% and 86% of ICC, respectively (not significant). Of all variants identified, the E6 variant 350G (L83V) and the E7 variant 822G were most frequently detected irrespective of histology and showed prevalence rates of 27% to 43% and 7% to 20%, respectively. No statistically significant differences in the prevalence of the E6 or E7 prototype sequences, any variants or multivariants in German women with < or = CIN 1, CIN 2/3 or ICC were found. PMID- 10389754 TI - Presence of high levels of MT1-MMP protein in fibroblastic cells of human invasive carcinomas. AB - Matrix metalloproteinase 2 (MMP2) activity is associated with the aggressiveness of human cancers. Therefore, the mechanisms regulating its activation are of great interest for a better understanding of malignant invasive processes. MT1 MMP, a membrane-bound MMP, is involved in the conversion of the latent form of MMP2 to the active one. In the present study, we have raised 3 monoclonal antibodies (Mabs) directed against 3 different epitopes of human MT1-MMP, which we used to investigate the expression and cellular localization of MT1-MMP protein in human carcinomas. MT1-MMP protein was present in all invasive carcinomas tested, and it was almost exclusively located to the stromal cells and not to cancer cells as previously reported, suggesting that MMP2 activation might be a peri-fibroblastic event. PMID- 10389755 TI - Neurotensin receptors in human neoplasms: high incidence in Ewing's sarcomas. AB - Receptors for regulatory peptides, such as somatostatin or vasoactive intestinal peptide (VIP), expressed at high density by neoplastic cells, can be instrumental for tumor diagnosis and therapy. Little is known about the expression of neurotensin receptors in human tumors. In the present study, 464 human neoplasms of various types were investigated for their neurotensin receptor content by in vitro receptor autoradiography on tissue sections using 125I-[Tyr3]-neurotensin as radioligand. Neurotensin receptors were identified and localized in tumor cells of 11/17 Ewing's sarcomas, 21/40 meningiomas, 10/23 astrocytomas, 5/13 medulloblastomas, 7/24 medullary thyroid cancers and 2/8 small cell lung cancers. They were rarely found in non-small cell lung cancers and breast carcinomas; they were absent in prostate, ovarian, renal cell and hepatocellular carcinomas, neuroendocrine gut tumors, pituitary adenomas, schwannomas, neuroblastomas and lymphomas. When present, the receptors bound with nanomolar affinity neurotensin and acetyl-neurotensin-(8-13), with lower affinity neuromedin N, diethylenetriamine penta-acetic acidneurotensin-(8-13) and SR 48692, but not neurotensin-(1-11). They were all of the NT1 type, without high affinity for levocabastine. Further, in 2 receptor-positive Ewing's sarcomas, neurotensin mRNA was detected by in situ hybridization techniques. Since neurotensin is known to stimulate cell proliferation, the presence of neurotensin receptors in human neoplasia may be of biological relevance, possibly as an integrative part of an autocrine feedback mechanism of tumor growth stimulation. PMID- 10389756 TI - Site-specific experimental metastasis patterns of two human breast cancer cell lines in nude rats. AB - Animal models for breast cancer metastasis are valuable tools for studying mechanisms of metastasis and for preclinical testing of anti-metastatic therapy regimens. Using MA-11 and MT-1, two oestrogen and progesterone receptor-negative human breast cancer cell lines, we developed new models in nude rats with patterns of experimental metastasis resembling those frequently observed in humans. MA-11 cells showed a clear preference for growth in the CNS. Fourteen of 15 animals injected with MA-11 cells into the left ventricle of the heart developed hind-leg paralysis, and tumours were observed in the spinal cord. MT-1 cells consistently exhibited bone/bone marrow metastases after intracardial injection, in addition to tumours in the brain and spinal cord. When injected into the cisterna magna, both cell lines gave rise to leptomeningeal neoplastic disease. Injection of MA-11 cells into the tibial bone marrow resulted in tumours in only 2 of 13 rats, whereas all animals injected with MT-1 cells developed tumours. Only 2 of 6 rats injected i.v. with MA-11 cells developed lung colonies compared with all 9 animals injected with MT-1 cells. Cell-surface expression of the following was examined: EGP2; MUC1; EGFr; E- and N-cadherin; the alpha2, alpha3, alpha5, beta1 and beta4 integrins; c-erb-B2; and N-CAM. c-erb-B2 was expressed in a higher percentage of the bone-metastasizing MT-1 cells than the MA 11 cells, whereas E-cadherin was expressed in MA-11 but not MT-1 cells. In animals injected with MA-11 and MT-1 cells in the left cardiac ventricle, treatment with cisplatin and doxorubicin did not improve survival. In summary, these clinically relevant animal models may be used for studies related to site specific growth and metastasis and for assessing effects of experimental therapy against human breast cancer. PMID- 10389758 TI - FGF7/KGF triggers cell transformation and invasion on immortalised human prostatic epithelial PNT1A cells. AB - Fibroblast growth factor 7 (FGF7/KGF) is synthesized exclusively by fibroblasts in normal tissues; it acts as a potent mitogen on epithelial cells, through interaction with the FGF7-specific receptor FGFR2/IIIb. To examine the importance of this growth factor both to prostate physiology and to prostate-cancer progression, we have tested the exogenous effect of FGF7. Thus, by mimicking the paracrine pathway (on proliferation, growth in soft agar and invasion) on the human prostatic epithelial cell line PNT1A positively checked for FGFR2/IIIb expression, FGF7 significantly enhanced cell proliferation at an optimal concentration of 7.5 x 10(-11) M, but no significant invasion or growth in soft agar were observed. To confirm FGF7 properties on human prostatic epithelial cells, we constitutively expressed FGF7 by transfecting PNT1A cells with FGF7 cDNA. The FGF7-transfected clones, PNT1A/ FGF7-T5 and PNT1A/FGF7-T6, were stable and expressed FGF7. Analysis of the FGF7-autocrine loop on the non-tumorigenic epithelial cells PNT1A showed acquired invasive potential in in vitro extracellular-matrix migration assays, specifically inhibited by an FGF7 neutralizing antibody, and over-expressed factors implicated in the migration process: the metalloproteinase MMP-1 and the plasminogen activator uPA. Taken together, these results demonstrate a role for FGF7 in triggering invasion of human prostatic epithelial cells. Furthermore, these FGF7-transfected clones exhibited functional and physiological differences from the original PNT1A cell line: anchorage-independent growth, growth in serum-free media and increased proliferation. These data confirm the oncogenic function of FGF7 in prostate progression potentially acting through paracrine and/or autocrine regulatory pathways. PMID- 10389757 TI - Antitumor activity and novel DNA-self-strand-breaking mechanism of CNDAC (1-(2-C cyano-2-deoxy-beta-D-arabino-pentofuranosyl) cytosine) and its N4-palmitoyl derivative (CS-682). AB - We have studied the antitumor activity and the novel DNA-self-strand-breaking mechanism of CNDAC (1-(2-Ccyano-2-deoxy-beta-D-arabino-pentofuranosyl)cytosine) and its N4-palmitoyl derivative (CS-682). In vitro, CS-682 showed strong cytotoxicity against human tumor cells comparable with that of CNDAC; both compounds displayed a similar broad spectrum. In vivo, however, orally administered CS-682 showed a more potent activity against human tumor xenografts than CNDAC, 5'-deoxy-5-fluorouridine, 5-fluorouracil and 2',2' difluorodeoxycytidine. Moreover, CS-682 was effective against various human organ tumor xenografts at a wide dose range and with low toxicity, and was effective against P388 leukemic cells resistant to mitomycin-C, vincristine, 5-fluorouracil or cisplatin in syngeneic mice. CNDAC, an active metabolite of CS-682, had a prolonged plasma half-life after repeated oral administrations of CS-682 but not after oral administrations of CNDAC itself. This difference may partially explain the higher antitumor activity of CS-682 relative to CNDAC. In both CNDAC- and CS 682-treated carcinoma cells, CNDAC 5'-triphosphate (CNDACTP) was generated and incorporated into a DNA strand. High performance liquid chromatography (HPLC) and mass spectrometric analysis of the nucleosides prepared by digestion of the DNA from the CNDAC-treated cells detected ddCNC (2'-Ccyano-2',3 '-didehydro-2',3 ' dideoxycytidine), which was shown to be generated only when the self-strand breakage of CNDACTP-incorporated DNA occurred. The cytotoxicity of CNDAC was completely abrogated by the addition of 2'-deoxycytidine and was low against cells with decreased deoxycytidine kinase. Our results suggest that CNDAC is converted to CNDACMP by deoxycytidine kinase and that the resulting CNDACTP incorporated into a DNA strand as CNDACMP may induce DNA-self-strand-breakage. This novel DNA-self-strand-breaking mechanism may contribute to the potent antitumor activity of CS-682. PMID- 10389759 TI - Regulation of spontaneous and TNF/IFN-induced IL-6 expression in two human ovarian-carcinoma cell lines. AB - Autocrine and paracrine production of interleukin-6 (IL-6) is considered to be involved in the ongoing proliferation of ovarian-cancer cells. In view of the variability in IL-6 expression between various ovarian-cancer cells, we questioned whether differences in IL-6-gene regulation might be observed in ovarian tumor cells with and without IL-6 expression. The CAOV-3 cell line spontaneously secreted IL-6, which was enhanced by tumor necrosis factor-alpha (877 +/- 89 vs. 8,452 +/- 1,762 pg/ml, x +/- sd, p < 0.01). The electrophoretic mobility shift assay (EMSA) demonstrated that basic IL-6 expression was associated with DNA binding of activator protein-1 (AP-1) and nuclear factor IL-6 (NF-IL6). Nuclear factor kappa-B (NF-KB), which consisted mainly of p65-NF-KB was induced in response to TNF-alpha stimulation. A2780 cells did not express IL-6, either spontaneously or after stimulation with TNF-alpha. EMSAs, showed spontaneous AP-1 but no NF-IL6 or NF-KB DNA binding. TNF-alpha stimulation enhanced AP-1 and induced NF-KB but no NF-IL6 DNA binding in these cells. NF-IL6 protein, however, was detected in nuclear extracts of these cells by Western blotting. In contrast, IL-6-promoter transfection studies showed no difference in promoter activation between CAOV-3 and A2780. This study reveals that differential IL-6-gene expression observed in ovarian-cancer cell lines is independent of NF-IL6 activation. PMID- 10389760 TI - Genetic analysis of sorted Hodgkin and Reed-Sternberg cells using comparative genomic hybridization. AB - Hodgkin and Reed-Sternberg (H and RS) cells are generally considered to be the neoplastic cells of Hodgkin's disease (HD); however, such cells are found only in a minority of HD lesions. Very few data have so far been published on the cytogenetic abnormalities in HD. An analysis of unknown genetic aberrations has only recently become possible through the use of comparative genomic hybridization (CGH), which is based on the competitive binding of tumor and control DNA to metaphase chromosomes. In order to exclude the reaction of non tumor cells, we used 100 sorted H-RS cells as the tumor DNA, then 100 sorted reactive T cells or B cells as the control DNA. We obtained the amplified DNA, using degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR). In addition, to confirm whether or not the lymphocytes in the background were reactive, we performed CGH with 100 sorted B cells and 100 sorted T cells. CGH was thus performed on 9 HDs, including 6 cases of mixed-cellularity (MC) sub-type and 3 cases of nodular-sclerosis (NS) sub-type. CGH of the B and T cells showed no genetic changes in any cases. In contrast, CGH of H-RS cells revealed both gains and losses of DNA in all 9 cases, and multiple changes were also observed. In situ hybridization showed an Epstein-Barr-virus infection in 5 cases of MC; however, no definite relationship was observed between the EBV infection and genetic changes. The most commonly observed genetic aberrations were a loss on 16q11/21 in 6 cases, a gain on 1p13 in 5 cases, and a gain on 7q35/36 in 5 cases. The large number of chromosomal alterations in HD suggests, therefore, that an increased degree of genetic instability play a role in the formation of H-RS cells. PMID- 10389761 TI - MUC1 isoform specific monoclonal antibody 6E6/2 detects preferential expression of the novel MUC1/Y protein in breast and ovarian cancer. AB - The products of the MUC1 gene are known to be highly expressed in human breast cancer cells. The best characterized MUC1 protein is a polymorphic, type 1 transmembrane molecule containing a large extracellular domain composed primarily of a variable number of 20 amino acid tandem repeats. We have recently identified a novel protein product of the MUC1 gene, the MUC1/Y protein, that is also a transmembrane protein but is devoid of the tandem repeat array and its immediate flanking sequences. To analyze its expression in tumor cells we generated monoclonal antibodies directed against the MUC1/Y extracellular domain (anti MUC1/Yex MAbs). Epitope mapping identified the MAb, 6E6, which recognized the MUC1/Y isoform with exquisite specificity- the repeat-array-containing MUC1 isoform could not compete out this immunoreactivity. A 30mer peptide which is unique for MUC1/Y and corresponds to the "join" region generated by the MUC1/Y specific splice, abrogated all 6E6 MAb immunoreactivity towards MUC1/Y. Immunoprecipitation of the MUC1/Y protein with 6E6 MAbs revealed that, in contrast with the proteolytic cleavage of the tandem-repeat-array-containing MUC1 isoform, MUC1/Y is not cleaved. Flow cytometry analyses using the 6E6 MAbs demonstrated that the MUC1/Y isoform is expressed on the cell surface of both MCF 7 breast cancer cells and malignant epithelial cells present in effusions obtained from breast and ovarian cancer patients. Our results unequivocally establish that the MUC1/Y protein is expressed on the surface of breast cancer cells and cells of other epithelial malignancies. The anti-MUC1/Y MAbs described here can target MUC1/Y expressing tumor cells in vivo and are likely to be important reagents both for epithelial tumor diagnosis and immunotherapy. PMID- 10389762 TI - Mitogen-activated protein kinase (MAPK) regulates the expression of progelatinase B (MMP-9) in breast epithelial cells. AB - Mitogen-activated protein kinases (MAPKs) play a major role in the mitogenic signal transduction pathway and are essential components of both growth and differentiation. Constitutive activation of the MAPK cascade is associated with the carcinogenesis and metastasis of human breast and renal cell carcinomas. The gelatinases B (MMP-9) and A (MMP-2) are 2 members of the matrix metalloproteinase (MMPs) family which are expressed in human cancers and thought to play a critical role in tumor cell invasion and metastasis. In a previous study, we have shown that EGF and amphiregulin upregulate MMP-9 in metastatic SKBR-3 cells but have no effect on MMP-2 secretion. We now investigated specific step(s) in EGF-induced signalling associated with regulation of cell proliferation and MMP-9 induction. EGF-induced signalling in SKBR-3 cells was blocked by relatively specific inhibitors either on ras (FPT inhibitor-1) or P13 kinase (Wortmannin) or by reduction in EGF-induced tyrosine kinase activity (RG 13022). Blocking these signalling pathways significantly inhibited of EGF-induced cell proliferation but only partially reduced in EGF-induced MMP-9 secretion. In contrast, when SKBR-3 cells were exposed to MEK inhibitor (PD 98059) or MAPK inhibitors (Apigenin or MAPK antisense phosphorothioate oligodeoxynucleotides), EGF-induced cell proliferation, MMP-9 induction and invasion through reconstituted basement membrane were significantly reduced. Our results suggest that interfering with MAPK activity may provide a novel means of controlling growth and invasiveness of tumors in which the signalling cascade is activated. PMID- 10389763 TI - Regulation of matrix metalloproteinase-2 (MMP-2) by hepatocyte growth factor/scatter factor (HGF/SF) in human glioma cells: HGF/SF enhances MMP-2 expression and activation accompanying up-regulation of membrane type-1 MMP. AB - Hepatocyte growth factor/scatter factor (HGF/SF) contributes to the malignant progression of human gliomas. We investigated the effect of HGF/SF on matrix metalloproteinase-2 (MMP-2), membrane type 1 matrix metalloproteinase (MT1-MMP) and tissue inhibitors of metalloproteinases (TIMPs), expressions of c-Met/HGF receptor-positive human glioblastoma cells. Treatment of U251 human glioblastoma cells with HGF/SF resulted in enhanced secretion of MMP-2 with an increased level of the active form. This was accompanied by enhanced expression (2.5-fold) of mRNA specific for MMP-2. The stimulatory effect of HGF/SF on MMP-2 expression did not occur in the presence of herbimycin A, a protein tyrosine kinase inhibitor. MT1 -MMP, a cell-surface activator of proMMP-2, was also up-regulated by HGF/SF in a dose-dependent manner. By contrast, the level of TIMP- 1 mRNAs was not altered significantly and that of TIMP-2 was reduced mildly by the HGF/SF treatment, suggesting that HGF/SF may eventually modulate a balance between MMP-2 and TIMPs in favor of the proteinase activity in the glioma cell microenvironment. HGF/SF also stimulated MMP-2 expression of other glioblastoma cell lines. Since glioblastomas frequently co-express HGF/SF and its receptor, our results suggest that HGF/SF might contribute to the invasiveness of glioblastoma cells through autocrine induction of MMP-2 expression and activation. PMID- 10389765 TI - Arsenic trioxide induces apoptosis of HPV16 DNA-immortalized human cervical epithelial cells and selectively inhibits viral gene expression. AB - Arsenic trioxide (As2O3), a major ingredient of arsenic compounds in traditional Chinese medicine, exhibits anti-acute promyelocytic leukemic activity. Considering that over 80% of human malignant tumors derive from epithelial cells, we studied the effect of As2O3 on HPV 16 DNA-immortalized human cervical epithelial cells (HCE16/3 cells) in vitro. As2O3 reduced HCE16/3 cell survival, induced apoptosis at a low concentration and selectively inhibited expression of viral early genes. This effect was evidenced by a reduction of cell viability in the MTT assay, G1 arrest and significant apoptosis upon flow-cytometric analysis, presence of apoptotic bodies, formation of DNA ladders upon gel electrophoresis and inhibition of viral early gene expression by RT-PCR and Western blot. There was a good correlation between cell apoptosis and viral early gene inhibition after As2O3 treatment, suggesting that induction of apoptosis of HCE16/3 cells by As2O3 treatment might be associated with down-regulation of viral oncogene expression. In conclusion, our findings indicate that As2O3 induces apoptosis of HCE16/3 cells, which may provide a new approach for treating HPV-associated tumors. PMID- 10389764 TI - Establishment, characterisation and partial cytokine expression profile of a new human osteosarcoma cell line (CAL 72). AB - Permanent human osteosarcoma cell lines are important tools for the study of bone cancer. As representative of an osteoblastic phenotype, they partly reflect their normal osteoblastic counterparts and, thus, may represent appropriate models to investigate the mechanisms involved in bone remodelling and in haematopoietic differentiation. In the present work, we describe a new human cell line, CAL 72, obtained from an osteosarcoma of the knee of a 10-year-old boy. These cells grow in continuous culture, and karyotypic analysis has revealed clonal abnormalities in number and structure, especially loss of chromosome Y. These cells exhibit morphological, immuno-histochemical and molecular characteristics of the osteoblastic lineage. Using RT-PCR, we have shown that the CAL 72 cell line expresses high levels of mRNA coding for several cytokines, such as G-CSF, GM CSF, IL-1beta and IL-6. In view of this expression profile, the CAL 72 phenotype appears to be closer to normal primary osteoblasts than other reported osteosarcomas. Moreover, these cells express mRNA for both HGF and its receptor c MET, suggesting that this autocrine loop might contribute to the invasiveness of the tumour from which CAL 72 originated. PMID- 10389766 TI - Adenoviral transduction of a cytosine deaminase/thymidine kinase fusion gene into prostate carcinoma cells enhances prodrug and radiation sensitivity. AB - Prostate tumor cells (PC-3) were transduced with defective, recombinant adenovirus containing a fusion gene encoding the Escherichia coli cytosine deaminase and herpes simplex virus type-1 thymidine kinase under the control of a cytomegalovirus promoter. Expression levels of the fusion protein were dependent on the multiplicity of infection used and incubation time following infection. PC 3 cells expressing this protein were sensitized to killing by the normally innocuous prodrugs 5-fluorocytosine and ganciclovir. In addition, radiation induced killing was enhanced in virally infected cells in the presence of the prodrugs. PMID- 10389768 TI - Functional reassessment of P16 variants using a transfection-based assay. AB - CDKN2A appears to be the major melanoma susceptibility gene, and is also mutated/deleted in sporadic tumours of various types including melanoma. Thus far most approaches to assessing the functionality of mutations in this gene have used in vitro methods such as CDK4 binding and kinase inhibition assays, with sometimes disparate conclusions about functional significance of some variants between studies. We have used a melanoma cell line (MM96L) with no functional p16, as the basis for a "semi-in vivo" transfection-based assay for exogenous p16 functionality based on the growth parameters of the cells and the behaviour of variant proteins after transfection of different CDKN2A cDNAs. Colony counts performed on these transfectants revealed that all but the wild type, + 24 bp ad A148T variants have a diminished ability to inhibit cell growth. All other variants detected either constitutionally in familial melanoma patients (I49T, R87P, G101W and V126D) or somatically in melanomas (N71S, and P81L), appeared functionally impaired in this assay. This diminution of function was independent of CDK4 and CDK6 binding ability. Furthermore, the predominant localization of these variants within the cell was different from that of wt p16. This mislocalization may provide an explanation for their lack of function, or alternatively, it may also be an indicator that the cells are processing unstable, misfolded p16 proteins. This novel assay for assessment of functionality of p16 variants may better reflect the role of some of these mutations in vivo, and as such is a useful adjunct to other in vitro assays. PMID- 10389767 TI - Coordinated modulation of the fibroblast growth factor dual receptor mechanism during transformation from human colon adenoma to carcinoma. AB - Basic fibroblast growth factor (bFGF) is dependent on heparan sulphate for its ability to activate the cell surface signal transducing receptor. We have investigated the FGF dual receptor mechanism in a novel model of the transformation from human colon adenoma to carcinoma in vitro. Reverse transcription-polymerase chain reaction showed that mRNA for FGF receptors 1 and 2 were expressed in both the adenoma and carcinoma cells whereas immunocytochemistry showed that the expression of the FGF R1 was reduced significantly in the carcinoma cells. We have reported previously that the composition and sequence of human colon adenoma and carcinoma heparan sulphate (HS) differ in a defined and specific manner. The functional significance of these changes was assessed by affinity co-electrophoresis, which showed that the affinity of adenoma HS for bFGF was 10-fold greater than that of the carcinoma HS (Kd 220 nM vs. 2493 nM, respectively). In addition, Northern studies of the expression of syndecan 1 and 4 mRNA showed that proteoglycan core protein expression was reduced significantly in the carcinoma cells. These findings were associated with a reduced biological response to bFGF in the carcinoma cells that could be partially reversed by the addition of exogenous heparin, suggesting that both the proteoglycan and signal transducing receptor control the cells' response to bFGF. PMID- 10389769 TI - Maximizing operating room utilization: a landmark study. PMID- 10389770 TI - Just when we thought we understood patient-controlled analgesia... PMID- 10389771 TI - An operating room scheduling strategy to maximize the use of operating room block time: computer simulation of patient scheduling and survey of patients' preferences for surgical waiting time. AB - Determining the appropriate amount of block time to allocate to surgeons and selecting the days on which to schedule elective cases can maximize operating room (OR) use. We used computer simulation to model OR scheduling. Inputs in the computer model included different methods to determine when a patient will have surgery (on-line bin-packing algorithms), case durations, lengths of time patients wait for surgery (2 wk is the median longest length of time that the outpatients [n = 367] surveyed considered acceptable), hours of block time each day, and number of blocks each week. For block time to be allocated to maximize OR utilization, two parameters must be specified: the method used to decide on what day a patient will have surgery and the average length of time patients wait to have surgery. OR utilization depends greatly on, and increases as, the average length of time patients wait for surgery increases. IMPLICATIONS: Operating room utilization can be maximized by allocating block time for the elective cases based on expected total hours of elective cases, scheduling patients into the first available date provided open block time is available within 4 wk, and otherwise scheduling patients in "overflow" time outside of the block time. PMID- 10389772 TI - The feasibility of gastrothoracic ventricular pacing during transesophageal echocardiography. AB - We evaluated whether ventricular pacing is possible using pacing electrodes attached to a transesophageal echocardiography (TEE) probe in 20 patients undergoing elective cardiovascular surgery. A bipolar pacing lead was fixed with silicone adhesive anteriorly to the TEE probe with the distal electrode 25 mm from the TEE probe tip. The TEE probe was positioned to obtain a transgastric short-axis view of the left ventricle. The distal or proximal electrode on the TEE probe was the cathode; the chest electrode placed at the V5 lead position was the anode. Gastrothoracic ventricular pacing (GVP) was performed at 100 bpm at 30 or 50-ms pulse duration. Transgastric ventricular pacing (TVP) was also attempted using both TEE probe electrodes alternately as cathode/anode. Maximal generator output was 32 mA. GVP with the distal electrode as cathode was successful in 75% and 80% of patients at 30- and 50-ms pulse durations and 23.3+/ 5.8 mA and 22.6+/-5.8 mA threshold currents, respectively. However, success rates (20% and 25%, respectively) were significantly lower with the proximal electrode as cathode using the same pulse durations and 14.4+/-5.3 mA and 16.7+/-6.8 mA threshold currents. The TVP success rate was significantly lower than that for GVP. With optimization, this system could become an available technique for intraoperative emergency ventricular pacing. IMPLICATIONS: Using an endocardial pacing lead attached to a transesophageal echocardiography probe, gastrothoracic ventricular pacing can be performed successfully without complications in 75%-80% of patients undergoing cardiovascular surgery. PMID- 10389773 TI - Platelet function and anesthetics in cardiac surgery: an in vitro and ex vivo study. AB - We studied the effects of the anesthetics commonly used in cardiac surgery on platelet function. Fentanyl, droperidol, succinylcholine, pancuronium, thiopental, and diazepam at therapeutic concentrations were tested for their in vitro effects on the expression of platelet membrane glycoproteins Ib and IIbIIIa (GpIb, GpIIb-IIIa) and of P-selectin in anticoagulated whole blood by flow cytometry. The expression of P-selectin was determined under basal conditions, after the incubation of blood with adenosine diphosphate (ADP) 10 micromol/L, and the stable prostaglandin endoperoxide analog U46619 1 micromol/L. No drug affected the expression of P-selectin in unstimulated and ADP- or U46619 stimulated platelets, with the exception of thiopental, which markedly decreased the U46619-induced expression of P-selectin. Thiopental concentration-dependently inhibited U46619-induced and ADP-induced platelet aggregation, with effects on U46619-induced aggregation at therapeutic concentrations. To assess ex vivo effects, the same platelet markers were also assessed in blood obtained from 10 patients undergoing elective coronary surgery. Compared with basal values, platelet response to U46619 was significantly reduced just after the administration of anesthetic drugs, and the effect persisted for 48 h after surgery. Our study suggests that, at therapeutic concentrations, thiopental inhibits U46619-induced platelet activation both in vitro and ex vivo. The mechanisms responsible of this effect, together with its clinical significance, require further investigation. IMPLICATIONS: Thiopental inhibited prostaglandin induced platelet activation at therapeutic concentrations both in vitro and ex vivo in cardiac surgical patients whereas adenosine diphosphate-induced activation was affected only at supratherapeutic drug concentrations. Thus, administration of sodium thiopental may contribute to the in vivo impairment of platelet function in patients undergoing elective cardiac surgery. PMID- 10389774 TI - A comparative study of hemodynamic and T-wave criteria for detecting intravascular injection of the test dose (epinephrine) in sevoflurane anesthetized adults. AB - This study was designed to determine the efficacy of heart rate (HR), systolic blood pressure (SBP), and changes in T-wave morphology in detecting intravascular injection of 15 microg of epinephrine (test dose) in sevoflurane-anesthetized adults. In addition, the testing threshold using the T-wave amplitude was derived. Ninety-six healthy patients were randomized to receive end-tidal sevoflurane 0.5%, 1%, or 2% and nitrous oxide 67% in oxygen (n = 32 for each sevoflurane concentration). Each group of patients was further randomized to receive 3 mL of 1.5% lidocaine plus 15 microg of epinephrine IV or 3 mL of saline IV (n = 16 each). HR, SBP, and T-wave amplitude were continuously monitored for 5 min after the IV injection of the study drug. None receiving IV saline and 15,15, and 14 patients receiving the IV test dose developed HR increases > or =10 bpm during 0.5%, 1%, and 2% sevoflurane, respectively. No patient receiving saline and all patients receiving the test dose developed SBP increases > or =15 mm Hg. T-wave amplitude decreased by >0.1 mV and by >25% in all patients receiving the IV test dose, and its magnitude was similar regardless of the sevoflurane concentrations. When 0.1-mV and 25% decreases in T-wave amplitude were considered as testing thresholds, 100% sensitivities and specificities were obtained. We conclude that a peak SBP increase > or =15 mm Hg and a decrease in T-wave amplitude > or =0.1 mV and > or =25% are more reliable than a HR increase > or =10 bpm for detecting intravascular injection of epinephrine-containing test dose during sevoflurane anesthesia. IMPLICATIONS: To determine whether an epidural catheter resides in a blood vessel, a standard test dose containing a local anesthetic and 15 microg of epinephrine is used. We found that, in sevoflurane anesthetized adult patients, a systolic blood pressure increase > or =15 mm Hg and a decrease in T-wave amplitude > or =0.1 mV and > or =25% in lead II, but not a heart rate increase > or =10 bpm, are reliable indicators for detecting intravascular injection. PMID- 10389775 TI - Propofol anesthesia enhances pressor response to ephedrine in patients given clonidine. AB - We studied the hemodynamic effects of ephedrine in patients with or without clonidine premedication during either isoflurane or propofol anesthesia. Forty adult patients were randomly assigned to one of two groups: 20 patients received famotidine 20 mg orally (control group) and 20 received clonidine 3 microg/kg and famotidine 20 mg orally (clonidine group). Within each group, 10 patients were then anesthetized with isoflurane and 10 with propofol. Hemodynamic measurements were taken at 1-min intervals for 10 min after a bolus injection of ephedrine 0.1 mg/kg. The magnitude of the maximal pressor response to ephedrine was no different whether patients without clonidine were anesthetized with isoflurane (increase 5+/-7 mm Hg) or propofol (3+/-9 mm Hg); however, this response was greater (P<0.05) with propofol (17+/-6 mm Hg) versus isoflurane (6+/-5 mm Hg) in patients given clonidine. The arterial blood pressure increase in clonidine premedicated patients with propofol anesthesia was the largest among the four subgroups. The heart rate response to ephedrine was not significant in patients anesthetized with isoflurane and was small but significant in those anesthetized with propofol. The present results, together with previous studies on the effect of ephedrine in patients medicated with clonidine, suggest that the interaction between clonidine and ephedrine is modulated by the anesthetic used. IMPLICATIONS: We evaluated the pressor response to ephedrine during isoflurane or propofol anesthesia with or without clonidine premedication. Our study suggests that, in anesthetized patients premedicated with clonidine, decreases in blood pressure may be easier to reverse with ephedrine with some types of anesthesia (e.g., propofol) than with others (e.g., isoflurane). PMID- 10389776 TI - Attenuation of endothelium-dependent dilation of pig pulmonary arterioles after cardiopulmonary bypass is prevented by monoclonal antibody to complement C5a. AB - We examined whether pulmonary endothelial dysfunction associated with cardiopulmonary bypass (CPB) may be mediated by complement C5a in pigs. Pigs were placed on normothermic CPB for 1 h with or without a previous administration of 1.6 mg/kg anti-C5a monoclonal antibody (MAb), then reperfused for 2 h. Pulmonary tissue myeloperoxidase activity was measured. Expression of nitric oxide synthase (NOS) was measured by reverse transcriptase polymerase chain reaction and Western blotting. Pulmonary arterioles approximately 100 microm in diameter were preconstricted with the thromboxane analog U46619 1 microM, and relaxation responses to adenosine diphosphate 10(-9)-10(-4) M, substance P 10(-12)-10(-6) M, and sodium nitroprusside 10(-9)-10(-4) M were examined in vitro by videomicroscopy. Relaxation to the endothelium-dependent dilators adenosine diphosphate and substance P was attenuated after CPB; this attenuation was prevented by the previous administration of MAb. Relaxation to sodium nitroprusside was not affected by CPB. Neutrophil sequestration, as measured by MPO activity, increased after CPB, either with or without MAb. Transcription of NOS was unchanged by CPB, but translation of constitutive NOS was decreased after CPB, and this decrease was prevented by a previous administration of MAb. We conclude that pig pulmonary endothelial dysfunction associated with CPB may be mediated by C5a. The mechanism may involve changes in NOS translation. IMPLICATIONS: In pigs, pulmonary endothelial dysfunction may occur after cardiopulmonary bypass due to product(s) of complement activation. PMID- 10389777 TI - Halothane attenuates nitroglycerin-induced vasodilation and a decrease in intracellular Ca2+ in the rat thoracic aorta. AB - Although halothane inhibits endothelium-mediated vasorelaxation, the sites of inhibition remain controversial. Because the cytosolic concentration of Ca2+ ([Ca2+]i) has crucial roles for tension development, we examined the effects of halothane on nitroglycerin-induced vasorelaxation from the standpoint of [Ca2+]i. Isolated spiral strips of rat thoracic aorta without endothelium were suspended for isometric tension recordings in a physiologic salt solution. Muscle contraction was evoked with 10(-8) M norepinephrine, followed by endothelium independent vasorelaxation with nitroglycerin 10(-7) and 10(-6) M. The effects of halothane 1.5% and 3% on nitroglycerin-induced vasorelaxation were evaluated along with the concomitant measurement of [Ca2+]i using fura-2-Ca2+ fluorescence. In other muscle strips, incremental doses of norepinephrine were administered during halothane exposure to induce contractions comparable to those without halothane. Nitroglycerin dose-dependently reduced norepinephrine-induced muscle contractions, but the decrease in [Ca2+]i reached a plateau at 10(-7) M, which indicates that nitroglycerin induced [Ca2+]i-dependent and [Ca2+]i-independent vasorelaxation. Both concentrations of halothane inhibited nitroglycerin-induced decreases in muscle tension and [Ca2+]i, not only when the same dose of norepinephrine was used for contraction during halothane exposure, but also at incremental doses of norepinephrine. In conclusion, halothane inhibits nitroglycerin-induced vasorelaxation partly by suppressing Ca2+ changes in the smooth muscle. IMPLICATIONS: We examined nitroglycerin-induced vasorelaxation in the rat thoracic aorta, along with the concomitant measurement of the cytosolic concentrations of Ca2+, and found that halothane attenuated endothelium independent vasorelaxation by suppressing Ca2+ dynamics in the smooth muscle. PMID- 10389778 TI - Dyspnea during interscalene block after recent coronary bypass surgery. PMID- 10389779 TI - Factors associated with blood loss and blood product transfusions: a multivariate analysis in children after open-heart surgery. AB - In this prospective cohort study of 548 children undergoing open-heart surgery, we evaluated demographic and perioperative factors to identify variables associated with perioperative blood loss and blood product transfusions. Using multivariate analysis, younger patient age was found to be the variable most significantly associated with bleeding and transfusions. Higher preoperative hematocrit, complex surgery, lower platelet count during cardiopulmonary bypass (CPB), and longer duration of deep hypothermic circulatory arrest were also significantly associated with bleeding and transfusion. Excessive postoperative chest tube (CT) drainage was associated with intraoperative bleeding. Independently associated variables accounted for 76% of the variability in CT output measured after 2 h in intensive care. Patients were subdivided into children aged < or =1 yr (infants) and children >1 yr; infants bled more intraoperatively (P<0.005); had greater cumulative CT output at 2, 6, 12, and 24 h (P<0.0001); and received more blood products (P<0.0001). Factors associated with bleeding and transfusions varied with patient age. Lower body core temperature during CPB was highly associated with blood loss and transfusions in infants, whereas resternotomy, preoperative congestive heart failure, and prolonged duration of CPB were significant factors associated with bleeding and transfusions in children >1 yr old. IMPLICATIONS: Knowledge of the factors associated with blood loss and blood product transfusions can help to identify children at risk of excessive bleeding after open-heart surgery. PMID- 10389780 TI - A controlled study of transesophageal echocardiography to guide central venous catheter placement in congenital heart surgery patients. AB - Transesophageal echocardiography (TEE) and central venous catheter (CVC) placement are often used during congenital cardiac surgery. Complications of CVC placement include cardiac perforation, inadvertent arterial placement, and erroneous hemodynamic data from unrecognized malposition. In this study, we used a prospective, randomized, controlled design to evaluate the use of TEE to guide depth of insertion and confirm superior vena cava cannulation, and to improve the percentage of correctly placed CVCs and reduce complications of CVC placement. One hundred forty-five patients were studied. Eighty patients were randomized to have subclavian vein insertion, 64 to have internal jugular insertion, and 1 to have external jugular insertion of CVC. TEE-guided CVC placement resulted in 100% correct placement when assessed by preoperative TEE, versus 86% in the control group (72 of 72 vs. 63 of 73; P = 0.01). There was no difference in correct placement between the two groups when assessed by postoperative chest radiograph (81.9% TEE versus 75.3% control; P = not significant). One significant complication, a superior vena cava perforation, occurred in the control group. Time to placement was 9.6 min in the TEE group versus 8.0 min in the control group (P = 0.015). IMPLICATIONS: Transesophageal echocardiography can be used to guide central venous catheter placement in congenital heart surgery. Central venous catheters that seem to be located high in the right atrium by chest radiograph in these patients are often actually in the superior vena cava and pose little risk of cardiac perforation. PMID- 10389781 TI - Quality of emergence from anesthesia and incidence of vomiting with remifentanil in a pediatric population. AB - We conducted a randomized trial to compare the incidence of vomiting and the quality of emergence from anesthesia associated with the use of remifentanil versus a nonopiate. It was expected that remifentanil would provide smoother emergence from anesthesia with a comparably low rate of vomiting. The study sample consisted of 115 pediatric patients undergoing dental restoration and extraction who were randomly assigned to the nonopiate or remifentanil groups based on their hospital admission numbers. The nonopiate patients received sufficient desflurane to prevent movement, typically 7%-9%. The remifentanil group received remifentanil 0.2 microg x kg(-1) x min(-1) and enough desflurane to prevent movement, typically 3.2%-3.6%. A trained postanesthesia care unit nurse, blinded to the anesthetic technique, assessed the quality of emergence and incidence of vomiting. Sixty-three patients received remifentanil and 52 received the nonopiate. The groups were not significantly different in either quality of emergence or incidence of vomiting. Remifentanil provided results comparable to a nonopiate with no increase in emesis. IMPLICATIONS: A randomized, controlled clinical trial of 115 patients undergoing dental restoration indicated that an anesthetic technique using remifentanil provided quality of emergence comparable to and no greater incidence of vomiting than a nonopiate technique. PMID- 10389782 TI - Premedication with midazolam delays recovery after ambulatory sevoflurane anesthesia in children. AB - We studied the effect of oral premedication with midazolam on the recovery characteristics of sevoflurane anesthesia in small children. In a randomized, double-blinded study, 60 children (1-3 yr, ASA physical status I or II) undergoing ambulatory adenoidectomy received either midazolam 0.5 mg/kg (Group M) or placebo (Group P) PO approximately 30 min before the induction of anesthesia. All children received atropine 0.01 mg/kg IV and alfentanil 10 microg/kg IV before the induction of anesthesia with sevoflurane up to 8 vol% inspired concentration in N2O 67% in O2. Tracheal intubation was facilitated with mivacurium 0.2 mg/kg. Anesthesia was continued with sevoflurane adjusted to maintain hemodynamic stability. In the postanesthesia care unit, predetermined recovery end points (emergence, recovery, discharge) were recorded. A pain/ discomfort scale was used to determine the quality of recovery. A postoperative questionnaire was used to evaluate the well-being of the patient at home 24 h after surgery. Emergence (spontaneous eye opening), recovery (full points on the modified Aldrete scale), and discharge were achieved later in Group M than in Group P (15+/-6 vs. 11+/-3 min [P = 0.002], 25+/-17 vs. 16+/-6 min [P = 0.01], and 80+/-23 vs. 70+/-23 min [P = 0.03]). Side effects, postanesthetic excitement, and analgesic treatment did not differ significantly between groups. At home, more children in Group P (30%) experienced disturbed sleep during the night compared with those in Group M (4%) (P = 0.007). IMPLICATIONS: In this randomized, double-blinded, placebo-controlled study, premedication with midazolam 0.5 mg/kg PO delayed recovery in children 1-3 yr of age after brief (<30 min) sevoflurane anesthesia. Except for more peaceful sleep at home, premedication did not affect the quality of recovery. PMID- 10389783 TI - Preoperative fasting in children. PMID- 10389784 TI - Voiding in patients managed with or without ultrasound monitoring of bladder volume after outpatient surgery. AB - The goal of this study was to determine whether recovery room monitoring of bladder volume would affect patient outcome after ambulatory surgery. Incidence of urinary retention and times to void and to discharge were compared in 161 patients managed with ultrasound bladder monitoring versus 173 controls without bladder monitoring. Urinary retention was diagnosed by clinical means or by ultrasound, confirmed by bladder catheterization. Patients were required to void or were catheterized before discharge. In the control patients without underlying risk factors for retention, median time to void was 95 min, and retention occurred in 0.8%, which was not significantly different from the ultrasound group (80 min and 0%, respectively). After hernia/anal surgery or spinal/epidural anesthesia, voiding was delayed (130 and 213 min), incidence of retention was increased (17% and 13%), and there was a trend toward earlier voiding (168+/-99 vs. 138+/-68 min) with bladder monitoring. We conclude that most patients at low risk of retention void within 3 h of outpatient surgery; their outcome is unaffected by bladder monitoring. After hernia/anal surgery and spinal/ epidural anesthesia, the likelihood of urinary retention is increased, and ultrasound monitoring facilitates deciding whether such patients should be catheterized. IMPLICATIONS: Incidence of bladder catheterization and urinary retention were compared in patients managed with and without ultrasound monitoring of bladder volume after outpatient surgery. Monitoring did not alter outcome in patients at low risk of retention, but it facilitated determining when to catheterize patients at high risk of retention (hernia/anal surgery, spinal/epidural anesthesia). PMID- 10389785 TI - Small-dose ketamine enhances morphine-induced analgesia after outpatient surgery. AB - Small-dose ketamine may enhance the analgesic effect of opiates. We studied the effect of IV coadministration of small-dose ketamine 50-100 microg/kg with morphine 50 microg/kg on postoperative morphine requirements and pain in 140 patients undergoing outpatient surgery. Midazolam 1-2 mg was administered in the holding area. Anesthesia was induced with propofol 2-2.5 mg/kg and was maintained with desflurane in a nitrous oxide/oxygen mixture. Patients received morphine 50 microg/kg with placebo (Group 1, n = 35) or ketamine 50 microg/kg IV (Group 2, n = 35), 75 microg/kg IV (Group 3, n = 35), or 100 microg/kg IV (Group 4, n = 35) 15 min before the end of the operation. Pain and drowsiness were assessed using visual analog scales on arrival in the recovery room, then every 15 min until the time of discharge to phase 2 recovery (phase 1 recovery). Morphine consumption in Groups 3 and 4 was approximately 40% less than that in the control group (91+/-9 and 89+/-8 microg/kg vs. 145+/-9 microg/kg; P<0.05 for both). Pain scores in Groups 3 and 4 were approximately 35% less than those in the control group at all time periods (P<0.0001 for all). There was no significant group difference in drowsiness scores. Small-dose ketamine 75-100 microg/kg IV, enhanced morphine induced analgesia after outpatient surgery. Simultaneous use of small doses of ketamine with morphine enhances the pain relief produced by morphine. IMPLICATIONS: Simultaneous use of small doses of ketamine with morphine enhances the pain relief produced by morphine. PMID- 10389786 TI - Nocturnal oxygenation during patient-controlled analgesia. AB - Patient-controlled analgesia (PCA) has become a standard modality for the management of postoperative pain, although anecdotal reports of excessive sedation and respiratory depression impugn its safety. To study the prevalence and severity of nocturnal hypoxemia, we measured arterial oxygen saturation (SpO2) continuously overnight in 32 postoperative patients who were receiving morphine via PCA. To evaluate the potential benefit of providing concurrent supplemental oxygen, the patients breathed oxygen-enriched air the night of surgery and room air the next night. Patients experienced more pain and consumed twice as much morphine the first night. However, breathing supplemental oxygen that night, the nocturnal mean SpO2 was 99%+/-1%, 94%+/-4% (P<0.001), and only four patients had periods of hemoglobin desaturation <90%. In contrast, breathing room air the subsequent night, the mean SpO2 was lower (94%+/-4%; P<0.001), and hypoxemia occurred more frequently and was more severe: 18 patients experienced episodes of SpO2 <90%, 7 patients experienced episodes of SpO2 <80%, and 3 patients experienced episodes of SpO2 <70%. One patient required resuscitation for profound bradypnea and cyanosis, but none suffered permanent sequelae. We conclude that when postoperative patients use PCA at night, hypoxemia can be substantial and oxygenation can be improved by providing supplemental oxygen. IMPLICATIONS: Oxygen saturation was measured postoperatively in patients using morphine patient-controlled analgesia. Substantial nocturnal hypoxemia occurred in half of the patients while they breathed room air. The severity of the hypoxemia was reduced when patients received supplemental oxygen. PMID- 10389787 TI - Ropivacaine epidural anesthesia and analgesia versus general anesthesia and intravenous patient-controlled analgesia with morphine in the perioperative management of hip replacement. Ropivacaine Hip Replacement Multicenter Study Group. AB - The aim of our study was to compare epidural anesthesia and analgesia (EDA) with ropivacaine versus general anesthesia followed by IV patient-controlled analgesia with morphine (GA/PCA) after hip replacement regarding pain, side effects, and discharge from the postanesthesia care unit. After ethics committee approval, randomization, and informed consent, 90 patients were enrolled. In Group EDA, epidural anesthesia (ropivacaine 10 mg/mL, 15-25 mL) was followed by an epidural infusion (2 mg/mL, 4-6 mL/h for 24 h, plus top-up doses of 6-10 mL for 48 h). GA/PCA patients received general anesthesia (isoflurane/N2O/fentanyl) followed by IV patient-controlled analgesia with morphine postoperatively. Pain was assessed by using visual analog scales (0-100 mm) at rest and during physiotherapy. Pain at rest was less in the EDA (n = 43) group than in the GA/PCA (n = 45) group (at 10 h: 11.8+/-12.9 vs. 28.4+/-17.1 [P< 0.001]; at 24 h: 14.3+/-11.7 vs. 24.0+/-17 [P<0.01]; in 48 h: 14.3+/-9.3 vs. 21.1+/-17.4 [P = 0.1]). Whereas EDA patients were deemed ready for discharge from the postanesthesia care unit earlier than GA/PCA patients (5.6+/-8.9 vs. 39.7+/-41.5 min), the actual discharge time was comparable. The median time for first passage of flatus was shorter in the EDA group than in the GA/PCA group (26 vs. 47 h). Nausea and vomiting were more common in the GA/PCA group than in the EDA group (16% vs. 28% and 11% vs. 22%, respectively), whereas hypotension (11% vs. 4%) and bradycardia (14% vs. 2%) were less frequent. Under the conditions of the present study, EDA with ropivacaine provided pain control after hip replacement superior to that provided by IV patient-controlled analgesia with morphine, particularly during the first 24 h. Both approaches to pain management were equally safe. IMPLICATIONS: Compared with general anesthesia and postoperative IV patient-controlled analgesia with morphine, epidural anesthesia and analgesia with the new local anesthetic ropivacaine enables patients to be discharged sooner from a postanesthesia care unit and provides superior pain relief during the first 24 h after hip replacement. PMID- 10389788 TI - Dexamethasone decreases epidural morphine-related nausea and vomiting. AB - The aim of our study was to compare the antiemetic effect of IV dexamethasone with saline control in preventing epidural morphine-related nausea and vomiting. Eighty patients requiring epidural anesthesia for abdominal total hysterectomy were enrolled in a randomized, double-blinded, and placebo-controlled study. At the end of surgery, all patients received epidural morphine 3 mg for relief of postoperative pain. Before the morphine injection, the dexamethasone group (n = 40) received IV dexamethasone 8 mg, whereas the saline group (n = 40) received IV saline. We found that the incidence of postoperative vomiting was 5% in the dexamethasone group and 25% in the saline group (P<0.05). The total incidence of nausea and vomiting was 16% in the dexamethasone group and 56% in the saline group (P<0.001). IV dexamethasone 8 mg significantly decreases the incidence of epidural morphine-related nausea and vomiting. IMPLICATIONS: We evaluated IV dexamethasone versus saline control in preventing epidural morphine-related nausea and vomiting in patients receiving epidural morphine for postoperative pain control. We found that IV dexamethasone significantly decreased the total incidence of nausea and vomiting after epidural morphine. IV dexamethasone may be a valuable treatment for preventing epidural morphine-related nausea and vomiting. PMID- 10389789 TI - Prolongation of nerve and epidural anesthetic blockade by bupivacaine in a lipid emulsion. AB - We assessed the effect of a lipid emulsion of bupivacaine on prolonging peripheral nerve and epidural anesthetic blockade in the rat. The intensity and duration of motor and sensory blockade produced by a single injection of aqueous solution (BPV-as) and lipid emulsion (BPV-em) preparations of 0.5% bupivacaine were evaluated by electrophysiological methods. Both preparations induced complete, reversible motor and sensory blockade after injection. The latency time to the maximal blockade and the duration of anesthetic blockade were more prolonged for BPV-em than for BPV-as. The increase in duration of maximal blockade was 1.4 times for nerve and 1.3 times for epidural anesthesia. Histological evaluation of spinal roots and spinal cord sections did not show any abnormalities or differences between animals injected with BPV-as and those injected with BPV-em. Pharmacokinetic studies showed lower plasma peak concentration and a longer elimination half-life for BPV-em than for BPV-as. Thus, BPV-em prolongs the effects of local anesthetics, allows a similar degree of blockade, and reduces the systems toxic effects of anesthetics compared with BPV-as. IMPLICATIONS: We assessed a lipid emulsion containing bupivacaine for peripheral nerve and epidural anesthetic blockade in the rat. The emulsion allowed a complete blockade, while increasing the duration of the anesthetic effect (by 30%-40%), compared with the standard bupivacaine aqueous solution. PMID- 10389791 TI - The safety and efficacy of intrathecal adenosine in patients with chronic neuropathic pain. AB - Adenosine and adenosine analogs decrease pain-like behavior in animal models of both acute nociceptive and neuropathic pain via adenosine receptor activation at spinal and/or supraspinal levels. This open study is the first in a series of intrathecal (IT) adenosine administration studied for the evaluation of efficacy and side effects in 14 patients. All had chronic neuropathic pain with tactile hyperalgesia and/or allodynia primarily of traumatic origin. The effects of IT adenosine (500 microg [n = 9] or 1000 microg [n = 5]) were evaluated. Approximate areas of tactile pain were mapped. Spontaneous and evoked pain (visual analog scale score 0-100) and tactile pain thresholds were assessed before and 60 min after injection. The injection caused transient pain (<60 min) in the lumbar region in five patients. There were no other side effects. Spontaneous and evoked pain was reduced (median score from 65 to 24 [P<0.01] and from 71 to 12 [P<0.01], respectively) in parallel with increased tactile pain thresholds in allodynic areas. Areas of tactile hyperalgesia/allodynia were reduced (median reduction 90%; P<0.001). Twelve patients experienced pain relief (median 24 h). We conclude that IT adenosine transiently causes lumbar pain in a subgroup of patients and may reduce various aspects of chronic neuropathic pain. IMPLICATIONS: This is the first series of patients with chronic neuropathic pain in which tolerability to spinal adenosine administration has been evaluated. A subset of patients reported transient low back pain as the only side effect. Spontaneous and evoked pain intensity decreased in most patients, an effect lasting for a median of 24 h. PMID- 10389790 TI - Pre- versus postformalin effects of ketamine or large-dose alfentanil in the rat: discordance between pain behavior and spinal Fos-like immunoreactivity. AB - The purpose of this animal investigation was to compare behavioral responses with spinal Fos-like immunoreactivity (FLI) after pre-versus postformalin administration of anesthetic doses of IV ketamine or alfentanil. Preformalin and postformalin injection (1.5% subcutaneously) treatment groups included IV saline control (1.5 mL/kg), ketamine (10 mg/kg), and alfentanil (170 microg/kg). In the behavioral study group, nociceptive behavior was evaluated 15-60 min after hindpaw formalin injection. In the spinal FLI study group, rats were perfused 2 h postformalin, and spinal cords were dissected, sliced at 30 microm, and processed by immunoperoxidase staining with an antibody against the Fos protein. Quantification and determination of the laminar distribution of Fos-labeled nuclei were performed at the L4-5 spinal level ipsilateral to formalin injection. Ketamine produced a selective preemptive analgesic effect in behavioral formalin experiments, yet failed to suppress spinal FLI. In contrast, alfentanil failed to demonstrate a selective preemptive analgesia in behavioral experiments, but did produce preemptive suppression of spinal FLI. Together with previous data from our laboratory, we conclude that behavioral analgesia and spinal Fos expression may be uncoupled under certain circumstances. IMPLICATIONS: In this study, we compared pain reduction produced by IV drugs (ketamine or alfentanil) with the ability to prevent injury-induced spinal cord changes. We measured pain behavior and spinal Fos protein after rats received ketamine or alfentanil before versus after formalin injection. Fos inhibition patterns did not clearly correlate with pain reduction, providing further evidence that Fos inhibition is not always predictive of behavioral analgesia. PMID- 10389792 TI - The spinal antinociceptive effects of a novel competitive AMPA receptor antagonist, YM872, on thermal or formalin-induced pain in rats. AB - Alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor antagonists have spinally mediated analgesic effects on acute nociception; however, their current formulations are not water-soluble and have toxic side effects. A new competitive AMPA antagonist, YM872 (2,3-dioxo-7-[1H-imidazol-1-yl] 6-nitro-1,2,3,4-tetrahydro-1-quinoxal inyl acetic acid) is water-soluble and may have fewer side effects. The purpose of this study was to investigate the analgesic effects of YM872 on both acute thermal and irritant-induced pain. Sprague-Dawley rats were implanted with chronic lumbar intrathecal catheters and were tested for their tail withdrawal response by the tail flick test and for their paw flinches by formalin injection after the intrathecal administration of YM872. The tail flick latency increased dose-dependently with a 50% effective dose (ED50) value of 1.0 microg. The number of flinches in both Phase 1 and Phase 2 of the formalin test decreased with increasing dose of YM872. ED50 values were 0.24 microg in Phase 1 and 0.21 microg in Phase 2. YM872 10 and 30 microg induced motor disturbance and flaccidity. In rats, the intrathecal administration of YM872 had analgesic effects on both acute thermal and formalin-induced nociceptions. Transient motor disturbance and flaccidity occurred only with large doses. YM872 may have potential in the clinical management of both acute and chronic pain. IMPLICATIONS: A novel alpha-amino-3-hydroxy-5-methylisoxazole-4 propionic acid (AMPA) receptor antagonist, YM872, may have an analgesic effect on both acute and chronic pain when administered intrathecally. PMID- 10389793 TI - Treatment of autonomic hyperreflexia in a quadriplegic patient by epidural anesthesia in the postoperative period. PMID- 10389794 TI - An unusual case of subcutaneous emphysema. PMID- 10389795 TI - Combined therapy with inhaled nitric oxide and intravenous vasodilators during acute and chronic experimental pulmonary hypertension. AB - Both inhaled nitric oxide (NO) and IV vasodilators decrease pulmonary hypertension, but the effects of combination therapy are unknown. We studied the response to inhaled NO (100 ppm) alone, IV vasodilator alone, and combined therapy during acute (U46619-induced) and chronic (monocrotaline-induced) pulmonary hypertension in the pentobarbital-anesthetized rat. Vasodilator doses were 1.0, 3.2, 10, and 32 microg x kg(-1) x min(-1) sodium nitroprusside (SNP); 50, 100, 150, 200, and 300 microg x kg(-1) x min(-1) adenosine; or 25, 50, 150, 200, and 300 ng x kg(-1) x min(-1) prostacyclin. In the absence of IV vasodilator therapy, inhaled NO decreased mean pulmonary artery pressure without decreasing mean systemic arterial pressure. In both acute and chronic pulmonary hypertension, the addition of inhaled NO to the largest dose of adenosine or prostacyclin, but not of SNP, decreased pulmonary artery pressure. Because inhaled NO and SNP activate guanylyl cyclase and adenosine and prostacyclin activate adenylyl cyclase, the results suggest that adding inhaled NO to a vasodilator not dependent on guanylyl cyclase may produce additional selective pulmonary vasodilation. IMPLICATIONS: In therapy of pulmonary hypertension, inhaled nitric oxide should produce additional selective pulmonary vasodilation when combined with a vasodilator whose mechanism of action is not dependent on cyclic guanosine 3',5'-monophosphate. PMID- 10389796 TI - Clonidine pretreatment inhibits stress-induced gastric ulcer in rats. AB - We studied the effects of clonidine (0.5 mg/kg) on hormonal stress response and antioxidant enzymes cold restraint-induced gastric lesions in rats. Rats in the study group were given 0.5 mg/kg intraperitoneal clonidine (n = 12), whereas the control group received 0.5 mL/kg intraperitoneal isotonic sodium chloride solution (n = 9). Animals were then subjected to immobilization at 4 degrees C in restraining devices for 4 h after a starvation period of 24 h. Gastric lesion index, gastric tissue malondialdehyde activity, and plasma cortisol concentrations were assayed. Histopathologic examination demonstrated a stress ulcer index of 3.17+/-0.92 mm in the clonidine group and 14.0+/-3.22 mm in the control group (P<0.05). The tissue malondialdehyde concentrations were slightly higher in the control group than in the clonidine group, but the differences were not statistically significant (P>0.05). Plasma cortisol levels were lower in the clonidine group (P<0.05). We concluded that clonidine attenuated the tissue damage and stress response in stress-induced gastric ulceration. IMPLICATIONS: Stressful circumstances can cause stomach ulcers, which can bleed, exposing patients to potentially life-threatening complications. In the present animal study, we showed that clonidine, a routinely available medication, may be useful in preventing stress-induced stomach ulcers. PMID- 10389797 TI - The effectiveness of rapidly infused intravenous fluids for inducing moderate hypothermia in neurosurgical patients. AB - Moderate hypothermia is often used for cerebral protection during anesthesia for cerebral aneurysm clipping. No reliable, rapid, and practical noncardiopulmonary bypass methods for the induction of hypothermia to core temperatures <34 degrees C have been reported. We assessed the effects of IV administration of chilled 5% albumin (5 mL/kg at 1-6 degrees C) on core temperature after surface cooling to approximately 34 degrees C. We calculated thermal distribution volume from the change in core temperature after the chilled fluid infusions. We also compared rapid administration (5 mL/kg over 30 min) with very rapid administration (5 mL/kg over 3-5 min). Chilled albumin 5 mL/kg infused over 5 min reduced core temperature by 0.6+/-0.1 degrees C. The same volume of chilled albumin infused over 30 min reduced core temperature by 0.4+/-0.1 degrees C. The calculated thermal distribution volume was less than one third of total body volume. Because the thermal distribution volume in these hypothermic patients was much lower than total body volume, the chilled IV fluids in this study were 3 times more effective in inducing hypothermia than suggested by a simple calculation. To achieve maximal effectiveness, however, chilled fluids must be administered very rapidly (>100 mL/min) to avoid heat gains in standard IV tubing that occur even with rapid administration. IMPLICATIONS: Chilled IV fluids can be much more effective for the induction of hypothermia than commonly assumed, but they must be administered very rapidly to avoid heat gains in IV tubing. PMID- 10389798 TI - The effects of sevoflurane and nitrous oxide on middle cerebral artery blood flow velocity and transient hyperemic response. AB - We studied the effects of sevoflurane, with and without nitrous oxide, on the indices of cerebral autoregulation (transient hyperemic response ratio and the strength of autoregulation) derived from the transient hyperemic response (THR) test. Twelve patients (ASA physical status I or II) aged 18-40 yr presenting for routine non-neurosurgical procedures were recruited. The middle cerebral artery blood flow velocity was continuously recorded using transcranial Doppler ultrasonography. Preinduction THR tests were performed before the patients were anesthetized with alfentanil, propofol, and vecuronium. End-tidal carbon dioxide concentration and mean arterial pressure (to within 10% with a phenylephrine infusion) were maintained at their preinduction values. THR tests were performed sequentially at the following end-tidal sevoflurane concentrations: 2.2% in oxygen, 3.4% in oxygen, 3.4% with 50% nitrous oxide in oxygen, and 2.2% with 50% nitrous oxide in oxygen. Neither 2.2% nor 3.4% sevoflurane significantly affected cerebral autoregulation. The addition of 50% nitrous oxide to the 2.2%, but not the 3.4%, concentration of sevoflurane increased middle cerebral artery blood flow velocity and decreased autoregulatory indices significantly. IMPLICATIONS: Transient hyperemic response is preserved during sevoflurane anesthesia but is significantly impaired when nitrous oxide is added to the lower concentration of sevoflurane (2.2%). These findings have implications for neurosurgical patients undergoing general anesthesia. PMID- 10389799 TI - The effects of nitrous oxide and oxygen on transient hyperemic response in human volunteers. AB - The aim of this study was to determine the effects of breathing 100% oxygen or 50% nitrous oxide in oxygen on the indices of cerebral autoregulation derived from the transient hyperemic response (THR) test in human volunteers. Data were analyzed from nine healthy subjects. Middle cerebral artery (MCA) blood flow velocity (FV) was measured by transcranial Doppler ultrasound, and the THR test was performed using 10-s compression of the common carotid artery. Continuous measurement of P(ETCO2) and expired fractions of oxygen (F(ETO2)) and nitrous oxide (F(ETN2O)) was established, and mean arterial pressure (MAP) was recorded at 2-min intervals. All measurements were performed while the volunteers were breathing room air and were repeated 10 min after achieving F(ETO2) >0.95 and 10 min after achieving F(ETN2O) 0.48-0.52. Two indices derived from the THR test, the transient hyperemic response ratio (THRR) and strength of autoregulation (SA), were used to assess cerebral autoregulation. P(ETCO2) and mean arterial pressure did not change significantly throughout the study period. Breathing 100% oxygen did not change MCA FV, THRR, or SA. Inhalation of nitrous oxide resulted in a marked and significant increase in the MCA FV (from 48+/-9 to 72+/-8 cm/s; mean +/- SD) and a significant decrease in the THRR (from 1.5+/-0.2 to 1.2+/-0.1) and the SA (from 1.0+/-0.1 to 0.8+/-0.1) (P<0.05 for all). We conclude that breathing 50% nitrous oxide in oxygen results in both a significant increase in MCA FV and impairment of transient hyperemic response. IMPLICATIONS: Our study suggests that nitrous oxide impairs cerebral autoregulation and may have implications for its use in neurosurgical anesthesia and for interpretation of the results from studies of anesthetics in which nitrous oxide is used in the background. PMID- 10389800 TI - The influence of the tonsillar gag on efficacy of seal, anatomic position, airway patency, and airway protection with the flexible laryngeal mask airway: a randomized, cross-over study of fresh adult cadavers. AB - We conducted a randomized, controlled, cross-over cadaver study to test the hypothesis that the efficacy of seal for ventilation and airway protection, anatomic position, and airway patency with the flexible laryngeal mask airway (FLMA) are altered by the application of a Boyle Davis (B-D) gag. We also determined the airway sealing pressure (ASP) at which the FLMA prevents aspiration when large volumes of fluid are placed above the cuff. We studied 20 adult cadavers (6-24 h postmortem). Efficacy of seal for ventilation and airway protection, anatomic position, and airway patency were determined with and without a B-D gag (two blade sizes: 8 and 10 cm) for the size 3, 4, and 5 FLMA in random order. Efficacy of seal for ventilation was determined by measuring the ASP at an intracuff pressure of 60 cm H2O. Efficacy of seal for airway protection was determined by flooding the mouth with 55-135 mL of water, reducing intracuff pressure until aspiration was detected fiberoptically and measuring ASP at this intracuff pressure. Anatomic position and airway patency were determined with a fiberoptic scope at an intracuff pressure of 60 cm H2O. In addition, in vivo compliance and ASP for the FLMA were measured in 10 cadavers and 10 paralyzed, anesthetized patients. Efficacy of seal for ventilation and airway protection, anatomic position, and airway patency did not change with the application of a gag for any mask size. The mean (range) ASP at which aspiration occurred when large volumes of fluid were placed above the cuff was 11 (7-15) cm H2O. The ASP for ventilation was always higher than the ASP for airway protection (P<0.0001). The FLMA had similar in vivo compliance and ASP in cadavers and anesthetized patients. We conclude that efficacy of seal for ventilation and airway protection, anatomic position and airway patency for the FLMA are unaffected by the application of a B-D gag in adults. ASP should be >15 cm H2O if there is a maximal risk of aspiration from above the cuff. IMPLICATIONS: The flexible laryngeal mask airway forms an effective seal for ventilation and protection of the airway that is unaffected by the application of a mouth gag that provides surgical access to the oropharynx. The efficacy of the seal should be >15 cm H2O if there is a maximal risk of aspiration from above the cuff. PMID- 10389801 TI - Nitrous oxide increases endotracheal cuff pressure and the incidence of tracheal lesions in anesthetized patients. AB - The pressure in air-filled endotracheal cuffs increases steadily throughout general anesthesia with nitrous oxide (N2O). High cuff pressures can be responsible for local ischemia, which may induce tracheal mucosal injury. In this study, cuff pressure was monitored in anesthetized patients, and postanesthesia endotracheal lesions were assessed by endoscopy. Sixty-five patients undergoing general anesthesia with tracheal intubation >1 h in duration were randomized into two groups. The endotracheal tube cuff was inflated to 30-40 cm H2O with air in Group 1 (n = 33) and with a gas mixture (N2O 50% in oxygen) in Group 2 (n = 32). At the time of tracheal extubation, a fiberoptic examination via the endotracheal tube was performed by an independent observer. Aspects of trachea at the level of cuff contact area were scored as 0 = normal, 1 = mucosal erythema or edema, 2 = mucosal erosion or hemorrhage, 3 = mucosal erosion or hemorrhage on both anterior and posterior tracheal walls. Cuff pressure increased throughout the procedure (P<0.01) in Group 1 and remained stable in Group 2. In Group 1, tracheal lesions in the area of the cuff were more frequent than they were in Group 2 (79% vs. 37%; P<0.001). Tracheal injury was correlated to cuff pressure (r = 0.62, P<0.001). No postoperative respiratory complication was observed in any patient. In patients anesthetized with N2O, the inflation of the tracheal tube cuff with a gas mixture of the same composition as the inhaled mixture can prevent excessive cuff pressure and reduce the incidence of tracheal injury. IMPLICATIONS: In patients anesthetized with nitrous oxide, the inflation of the tracheal tube cuff with a gas mixture of the same composition as the inhaled mixture can prevent excessive cuff pressure and reduce the incidence of tracheal injury. PMID- 10389802 TI - The relaxant effect of propofol on guinea pig tracheal muscle is independent of airway epithelial function and beta-adrenoceptor activity. AB - Airway epithelium and vascular endothelium modulate the tension of the underlying smooth muscle by releasing relaxing factors such as prostanoids and nitric oxide (NO). We investigated whether the relaxant effect of propofol on airway smooth muscle is dependent on airway epithelial function. Tracheal spirals of female guinea pigs were mounted in water-jacketed organ baths filled with Krebs bicarbonate buffer aerated with 95% O2 and 5% CO2 at 37 degrees C. Changes in isometric tension of the specimens were measured with a force-displacement transducer and recorded with a polygraph. Propofol (10(-4) to 10(-3) M) inhibited carbachol (CCh)-, histamine (HA)-, or endothelin-1-induced contractions of the muscles in a dose-dependent manner. Neither mechanical removal of the epithelial layer, chemical inhibition of epithelial synthesis of prostanoids, nor NO affected the relaxant effect of propofol on CCh- or HA-induced tracheal contraction. Furthermore, the blockade of beta-adrenoceptors did not change the relaxant effect of propofol. These results indicate that the relaxant effect of propofol on the airway smooth muscle is independent of the epithelial function or beta-adrenoceptor activity. Propofol is an excellent anesthetic for patients with hyperreactive airways in which the epithelial layer is damaged. IMPLICATIONS: Airway epithelium, as well as vascular endothelium, plays an important role in modulating the baseline tone and reactivity of underlying smooth muscle. We investigated, in vitro, whether the relaxant effect of propofol on airway smooth muscle is dependent on airway epithelial function. We suggest that propofol relaxes airway smooth muscle independently of the epithelial function. PMID- 10389803 TI - Ondansetron pretreatment to alleviate pain on propofol injection: a randomized, controlled, double-blinded study. AB - We used a randomized, controlled, double-blinded design to study the effect of ondansetron (OND) pretreatment on the pain produced by the IV injection of propofol. Eighty patients were randomly assigned to one of two groups: Group I received 2 mL of IV 0.9% saline pretreatment, and Group II received OND (4 mg in 2 mg/mL solution) pretreatment in the dorsum of the hand, followed by propofol 1 min later. Pain was reduced significantly in the OND group (P<0.05). Approximately one third of the patients in each group had myoclonic movements or skin rashes in the limb that received propofol. We conclude that the OND pretreatment may be used to reduce the incidence of pain on injection of propofol and to prevent postoperative nausea and vomiting. IMPLICATIONS: In a double blinded, controlled study, IV ondansetron (4 mg) pretreatment was used to alleviate pain on injection of propofol. Ondansetron was successful in relieving pain without any adverse effect in a significant number of patients. PMID- 10389804 TI - The prophylactic effect of dexamethasone on postoperative nausea and vomiting in women undergoing thyroidectomy: a comparison of droperidol with saline. AB - The aim of this study was to evaluate the prophylactic effect of dexamethasone on postoperative nausea and vomiting (PONV) in women undergoing thyroidectomy. Droperidol and saline served as controls. One hundred twenty women (n = 40 in each of three groups) undergoing thyroidectomy under general anesthesia were enrolled in this randomized, double-blinded, placebo-controlled study. Immediately before the induction of anesthesia, Group 1 received IV dexamethasone 10 mg, whereas Groups 2 and 3 received IV droperidol 1.25 mg and saline, respectively. We found that both dexamethasone and droperidol significantly decreased the total incidence of PONV compared with saline, with an incidence of 32%, 35%, and 76%, respectively (P<0.01; Group 1 versus Group 3, Group 2 versus Group 3). Patients who received droperidol, however, reported a higher intensity of sore throat and a more frequent incidence of restlessness than those who received dexamethasone. We conclude that, although both dexamethasone and droperidol are effective as prophylactic antiemetics in women undergoing thyroidectomy, droperidol produces more side effects. IMPLICATIONS: We compared the prophylactic administration of dexamethasone to prevent nausea and vomiting with droperidol and saline in women undergoing thyroidectomy. Both dexamethasone and droperidol significantly reduced postoperative nausea and vomiting, but droperidol produced more side effects, which suggests that dexamethasone is a useful treatment in these patients. PMID- 10389805 TI - The effects of clonidine premedication on sevoflurane requirements and anesthetic induction time. AB - We assessed the effects of oral clonidine preanesthetic medication (4.5 microg/kg) on the vital capacity rapid-inhalation anesthetic induction time (VCRII time) and minimum alveolar anesthetic concentration (MAC) to prevent a response to a verbal command in 50% of patients (MAC-Awake) by its hypnotic effect, and on MAC-Skin incision for the analgesic effect in patients anesthetized with sevoflurane. We studied 104 adult patients (control group: n = 52, clonidine group: n = 52) aged 30-48 yr scheduled to undergo general anesthesia. Fifty-two patients received oral clonidine 4.5 microg/kg 1.5 h before arrival in the operating room (clonidine group). The patients exhaled to residual volume and took three vital capacity breaths of 5% sevoflurane in oxygen. The VCRII time was defined as the time interval between the initiation of the VCRII and the disappearance of the response to verbal command. Anesthesia was maintained with sevoflurane in oxygen and air. The end-tidal (ET) sevoflurane concentration reached a predetermined value, then the ratio of predetermined ET to inspiratory concentration was maintained at > or =0.95 for at least 15 min before skin incision. After skin incision, the patients were observed for gross purposeful muscular movements. MAC was defined as the average of the cross-over midpoints in each cross-over. After maintaining the ET sevoflurane concentration for 15 min, patients were judged to be awake or asleep. Average times for VCRII using 5% sevoflurane were achieved in 44+/-11 s (mean +/- SD) and 27+/-6 s in the control and clonidine groups, respectively (P = 0.0001). MAC-Awake values of sevoflurane were 0.66%+/-0.03% and 0.35%+/-0.02% (P = 0.0001), and MAC-Skin incision values were 1.97%+/-0.19% and 1.29%+/-0.13% (P = 0.0001) in the control and clonidine groups, respectively. These results suggest that clonidine may have a more potent hypnotic effect than analgesic effect. IMPLICATIONS: Oral clonidine preanesthetic medication (4.5 microg/kg) significantly reduces vital capacity rapid inhalation anesthetic induction time and minimum alveolar anesthetic concentration awake for sevoflurane. PMID- 10389806 TI - Ventilatory responses to acute and sustained hypoxia during sevoflurane anesthesia in women. AB - We investigated the isocapnic hypoxic (i.e., pulse oximetry monitored arterial saturations 70%-75%) ventilatory response (HVR) for 20 min in the awake state and during sevoflurane anesthesia at an end-tidal concentration of 1.6% in eight healthy (ASA physical status I) women. Our aims were to determine if a prolonged isocapnic hypoxic period during sevoflurane anesthesia showed a biphasic response pattern (i.e., an initial acute HVR followed by a decline to a lowered sustained HVR) and, if so, to quantify to what extent the acute and sustained HVRs were depressed by anesthesia. The study was conducted before laparoscopic gynecological surgery. Pneumotachography and in-line infrared capnography were used. The pattern of awake biphasic HVR was maintained during anesthesia but was depressed during both the acute and the sustained phases by 60% and 70%, respectively. Further, HVR during anesthesia was accomplished by an increase in respiratory rate, in contrast to an increase in tidal volume in the awake state. In conclusion, sevoflurane anesthesia at 1.6% depresses HVR in women, but the biphasic response is maintained. IMPLICATIONS: Acute and sustained hypoxic ventilatory responses were investigated in eight women before and during sevoflurane anesthesia. A biphasic ventilatory response was persistent but blunted during anesthesia. PMID- 10389807 TI - The effects of increasing concentrations of desflurane on systemic oxygenation during one-lung ventilation in pigs. AB - During one-lung ventilation (OLV), hypoxic pulmonary vasoconstriction reduces venous admixture and attenuates the decrease in arterial O2 tension by diverting blood from the nonventilated to the ventilated lung. In vitro, increasing concentrations of desflurane depresses hypoxic pulmonary vasoconstriction in a dose-dependent manner. Accordingly, we investigated the effects of increasing concentrations of desflurane on oxygenation during OLV in vivo. Thirteen pigs (25 30 kg) were anesthetized (induction: propofol 2-3 mg/kg IV; maintenance: N2O/O2 50%/50%, desflurane 3%, propofol 50 microg x kg(-1) min(-1), and vecuronium 0.2 mg x kg(-1) x h(-1) IV), orotracheally intubated, and mechanically ventilated. After placement of femoral arterial and thermodilution pulmonary artery catheters, a leftsided, 28F, double-lumen tube was placed via tracheotomy. After double-lumen tube placement, N2O and desflurane were discontinued, propofol was increased to 200 microg x kg(-1) x min(-1), and the fraction of inspired oxygen was adjusted at 0.8. Anesthesia was then continued in random order with desflurane 5%, 10%, or 15% end-tidal concentrations while propofol was discontinued. Whereas mixed venous PO2, mean arterial pressure, cardiac output, and shunt fraction decreased in a dose-dependent manner, PaO2 remained unchanged with increasing concentrations of desflurane during OLV. These findings indicate that, in vivo, increasing concentrations of desflurane do not necessarily worsen oxygenation during OLV. IMPLICATIONS: Oxygenation during one-lung ventilation depends on reflex vasoconstriction in the nonventilated lung. In vitro, desflurane inhibits this reflex dose-dependently. Our results indicate that, in vivo, this does not necessarily translate to dose-dependent decreases in oxygenation during one-lung ventilation. PMID- 10389808 TI - The effects of cimetidine, ranitidine, and famotidine on human neutrophil functions. AB - Neutrophil functions, which play an important role in the antibacterial host defense system, are inhibited by various anesthetics and surgical procedures. Histamine H2-receptor antagonists are perioperatively used as a prophylaxis against acid aspiration syndrome or stress ulceration. We examined the effect of cimetidine, ranitidine, and famotidine, at clinically relevant concentrations and at 10 and 100 times this concentration, on several aspects of human neutrophil function using an in vitro system. The three H2-receptor antagonists did not impair neutrophils' chemotaxis or phagocytosis. Cimetidine and famotidine inhibited superoxide (O2-) and hydrogen peroxide (H2O2) production of the neutrophils in a dose-dependent manner, although the inhibitory effects were minimal. In contrast, ranitidine failed to change O2- or H2O2 production of neutrophils. The three H2-receptor antagonists did not scavenge these reactive oxygen species generated by the xanthine-xanthine oxidase system. The increase in intracellular calcium concentrations in neutrophils by a stimulant were dose dependently attenuated with cimetidine or famotidine. This decreasing effect of the drugs on [Ca2+]i in neutrophils may represent one of mechanisms responsible for inhibition of reactive oxygen species generation. IMPLICATIONS: Neutrophils play a pivotal role in the antibacterial host defense system and tissue injury. We found that cimetidine and famotidine slightly reduced the O2- or H2O2 production of neutrophils in a dose-dependent manner, although ranitidine failed to do so. At least ranitidine does not seem to have any deleterious effect on neutrophil function, which is clearly an important consideration in its use in severely ill patients. PMID- 10389809 TI - Volatile anesthetics reduce calcium current in parasympathetic neurons from bullfrog hearts. AB - Although the autonomic nervous system regulates cardiac function, the cellular mechanism(s) of general anesthetics on the activities of parasympathetic neurons have not been directly assessed. We therefore studied the volatile anesthetic actions on the Ca2+ current of parasympathetic neurons isolated from bullfrog hearts. Neurons were enzymatically isolated from the interatrial septum of bullfrog heart and maintained in a short-term tissue culture. The Ca2+ current was recorded with a whole-cell voltage-clamp method under a Na+, K+ -free condition. Isoflurane (2.5 vol%) and sevoflurane (5.0 vol%) reduced the peak amplitude of the Ca2+ current (to 79% and 72% of control, respectively) without changing the reversal potential. The curve-fit analysis of the inactivation kinetics revealed that isoflurane and sevoflurane accelerated the inactivation of the current and that isoflurane shifted the midpoint of the steady-state inactivation curve of the Ca2+ current toward negative by 13.6 mV. The results indicate that volatile anesthetics reduce the Ca2+ current of parasympathetic neurons and modify the inactivation kinetics. IMPLICATIONS: The anesthetic reduction of the Ca2+ current of parasympathetic neurons can induce a decrease of acetylcholine release from the post-ganglionic endings. These findings, in part, account for the anesthetic attenuation of the vagal efferent activities observed in humans and experimental animals. PMID- 10389811 TI - The complexity of tracheal intubation using rigid fiberoptic laryngoscopy (WuScope). PMID- 10389810 TI - Enantioselectivity of thiopental distribution into the central neural tissue of rats: an interaction with halothane. AB - Thiopental is a racemate. In this study, we examined whether thiopental total body clearance and its distribution into central nervous system (CNS) tissue of rats was enantioselective. Rats, either anesthetized with halothane or conscious and restrained, were infused to stepwise steady-state targets of 5, 10, and 20 microg/mL thiopental by computer-controlled infusions. Serial arterial plasma and steady-state samples of brain and spinal cord were assayed enantiospecifically for thiopental. In both groups, concurrent total and unbound plasma concentrations of S-thiopental were approximately 10%-20% higher than those of R thiopental, corresponding to its higher clearance. CNS tissue concentrations of S thiopental were approximately 20% higher than those of R-thiopental. Spinal cord to plasma distribution coefficients were approximately 2 x those in the brain, with relative distribution coefficients approximately 10% greater for R thiopental in both tissues. Plasma concentrations and distribution coefficients of both enantiomers were approximately 10%-20% lower in the halothane anesthetized group, with a slightly greater effect on R-thiopental distribution. We conclude that the total body clearance of R-thiopental > S-thiopental, that halothane enantioselectively reduces the relative uptake of R-thiopental into brain tissue, and that composition is important in determining the CNS tissue concentrations of thiopental. The reported higher potency of S-thiopental did not seem to be due to its greater distribution into CNS tissues. IMPLICATIONS: Because thiopental is a mixture of two forms (termed R-and S-enantiomers), correct interpretation of its distribution into, and clearance from, the body requires knowledge about both enantiomers. In this study, performed in rats, we showed that the two enantiomers of thiopental differed significantly, with the R enantiomer having the preferred profile. PMID- 10389812 TI - A new skin-protective sphygmomanometer cuff. PMID- 10389813 TI - The effects of residual neuromuscular blockade and volatile anesthetics on the control of ventilation. PMID- 10389814 TI - Mainstem bronchial obstruction during laparoscopic fundoplication. PMID- 10389815 TI - Measurement of patient satisfaction. PMID- 10389816 TI - Continuing inflammation does enhance spinally mediated antinociception by neostigmine in the rat model. PMID- 10389817 TI - Pharyngoscopic views. PMID- 10389818 TI - Use of a prototype flexible lighted catheter for guided tracheal intubation through the intubating laryngeal mask. PMID- 10389819 TI - Hyperventilation, hyperoxia, and cerebral oxygenation after traumatic brain injury. PMID- 10389820 TI - Oxygenation may improve with time during one-lung ventilation. PMID- 10389821 TI - The effects of propofol, isoflurane, and sevoflurane on oxygenation during one lung ventilation. PMID- 10389822 TI - A new class for modified Mallampati sign. PMID- 10389823 TI - Carbon dioxide tracing for face mask disconnects. PMID- 10389824 TI - Maximal intracuff volumes for the cuffed oropharyngeal airway in adults. PMID- 10389825 TI - Pharyngeal obstruction of a reinforced orotracheal tube. PMID- 10389826 TI - A potential hazard with nondisposable pulse oximeter probes. PMID- 10389827 TI - Sialadenopathy with the laryngeal mask airway. PMID- 10389828 TI - Continuous autotransfusion in a Jehovah's witness undergoing coronary artery bypass grafting. PMID- 10389829 TI - Sore throat and pharyngeal trauma after extratracheal airway placement: does the literature help practitioners? PMID- 10389830 TI - Postdural puncture headaches in special patient populations. PMID- 10389831 TI - Concept not new or recommended. PMID- 10389832 TI - Fully dilated, nonreactive pupils during cardiac anesthesia. PMID- 10389833 TI - Epidural falsely blamed for lower extremity temperature disparity. PMID- 10389834 TI - Bullard extender use with the Bullard laryngoscope. PMID- 10389835 TI - Additional Bullard tips. PMID- 10389836 TI - Bullard laryngoscope: keeping its act together. PMID- 10389837 TI - More tips for users of the Bullard laryngoscope. PMID- 10389838 TI - Comparative genetics of type 1 diabetes and autoimmune disease: common loci, common pathways? AB - Genome-scale analysis in type 1 diabetes has resulted in a number of non-major histocompatibility complex loci of varying levels of statistical significance. In no case has a specific gene been proven to be the source of genetic linkage at any candidate locus. Comparative analysis of the position of loci for type 1 diabetes with candidate loci from other autoimmune/inflammatory diseases shows considerable overlap. This supports a hypothesis that the underlying genetic susceptibility to type 1 diabetes may be shared with other clinically distinct autoimmune diseases such as systemic lupus erythemastosus, multiple sclerosis, and Crohn's Disease. PMID- 10389839 TI - Insulin-induced insulin receptor substrate-1 degradation is mediated by the proteasome degradation pathway. AB - Insulin receptor substrate (IRS) proteins are important intracellular molecules that mediate insulin receptor tyrosine kinase signaling. A decreased content of IRS proteins has been found in insulin-resistant states in animals, humans, and cultured cells under various conditions. However, the molecular mechanism that controls cellular levels of IRS proteins is unknown. We report that chronic insulin treatment induces the degradation of IRS-1, but not IRS-2, protein in cultured cells. The insulin-induced degradation of IRS-1 can be prevented by pretreatment with lactacystin, a specific inhibitor for proteasome degradation. These data demonstrate, for the first time, that insulin-induced degradation of IRS-1 is mediated by the proteasome degradation pathway. IRS-2 can escape from the insulin-induced proteasome degradation, suggesting the existence of specific structural requirements for this degradation process. PMID- 10389840 TI - No correlation of plasma cell 1 overexpression with insulin resistance in diabetic rats and 3T3-L1 adipocytes. AB - Membrane glycoprotein plasma cell 1 (PC-1) has been shown to be increased in type 2 diabetes and involved in insulin resistance through inhibiting the insulin receptor tyrosine kinase, which was demonstrated using cultured breast cancer cells. However, other reports have shown contradictory results in Chinese hamster ovary cells and in vitro kinase assay. Thus, we considered it necessary to investigate the effect of PC-1 using highly insulin-sensitive cells. Here, we used two of the following approaches: 1) investigating PC-1 expression levels in insulin-responsive tissues in rat models of diabetes and 2) overexpressing PC-1 in 3T3-L1 adipocytes. We found that PC-1 was highly expressed in insulin responsive tissues, such as liver and adipose tissue, in normal rats. However, high-fat feeding or streptozotocin-induced diabetes did not change its expression levels in liver, adipose tissue, and skeletal muscle. Thus, PC-1 expression levels were not associated with high-fat-diet-induced insulin resistance or hyperglycemia. Although PC-1 was increased in adipose tissue in Zucker fatty rats (protein level, by 50%; mRNA level, by 90%), its expression levels in liver and skeletal muscle, tissues that are more responsible for whole body glucose metabolism than adipose tissue, did not significantly differ from those in normal rats. Next, we overexpressed PC-1 in 3T3-L1 adipocytes using an adenovirus transfection system. PC-1 expression was markedly increased to a level 16-fold greater than that in normal human adipose tissue, which is higher than the previously reported levels in diabetic patients. However, insulin-induced tyrosine phosphorylation of the insulin receptor and insulin receptor substrate 1, activation of phosphatidylinositol 3-kinase, and glucose uptake were not affected by PC-1 overexpression. These results strongly suggest that increased PC 1 expression is not causally related to insulin resistance. PMID- 10389841 TI - Activation of the sphingomyelinase/ceramide signal transduction pathway in insulin-secreting beta-cells: role in cytokine-induced beta-cell death. AB - Activation of the sphingomyelin/ceramide pathway may mediate interleukin-1 induced beta-cell death (Welsh, N: Interleuken-1beta-induced ceramide and diacylglycerol generation may lead to activation of the c-Jun NH2-terminal kinase and the transcription factor ATF-2 in the insulin-producing cell line RINm5F. J Biol Chem 271: 8307-8312, 1996). In this report, we have examined this pathway in more detail. Culture of beta-TC3 cells with 25 micromol/l ceramide analogs (N acetyl- and N-hexanoylsphingosine) for 72 h did not significantly affect glucose- and carbachol-induced insulin secretion. Dihydroceramide (N-acetyl- or N hexanoylsphinganine), a structurally similar analog, had no effect on agonist induced secretion. However, ceramide analogs both time- and dose-dependently decreased cell viability, while the dihydroceramide analog had no effect. The ceramide effect on cell viability mimicked the effect of the cytokines TNF-alpha, IL-1beta, and IFN-gamma, reported stimulators of sphingomyelin hydrolysis. Cytokines, however, failed to stimulate sphingomyelin metabolism. Furthermore, using two different methods to quantitate ceramide, cytokines failed to cause an increase in beta-cell ceramide content versus unstimulated or time-matched vehicle controls. Taken together, these data suggest that although ceramide analogs mimic the cytotoxic effect of cytokines, activation of the sphingomyelin/ceramide signaling pathway is not involved in cytokine-induced beta cell death. PMID- 10389842 TI - Why do microencapsulated islet grafts fail in the absence of fibrotic overgrowth? AB - The survival of microencapsulated islet grafts is limited, even if capsular overgrowth is restricted to a small percentage of the capsules. In search of processes other than overgrowth contributing to graft failure, we have studied the islets in non-overgrown capsules at several time points after allotransplantation in the rat. All recipients of islet allografts became normoglycemic. Grafts were retrieved at 4 and 8 weeks after implantation and at 15.3 +/- 2.3 weeks postimplant, 2 weeks after the mean time period at which graft failure occurred. Overgrowth of capsules was complete within 4 weeks postimplant, and it was usually restricted to <10% of the capsules. During the first 4 weeks of implantation, 40% of the initial number of islets was lost. Thereafter, we observed a decrease in function rather than in numbers of islets, as illustrated by a decline in the ex vivo glucose-induced insulin response. At 4 and 8 weeks postimplant, beta-cell replication was 10-fold higher in encapsulated islets than in islets in the normal pancreas, but these high replication rates were insufficient to prevent a progressive increase in the percentage of nonviable tissue in the islets. Necrosis and not apoptosis proved to be the major cause of cell death in the islets. The necrosis mainly occurred in the center of the islets, which indicates insufficient nutrition as a major causative factor. Our study demonstrates that not only capsular overgrowth but also an imbalance between beta-cell birth and beta-cell death contributes to the failure of encapsulated islet grafts. Our observations indicate that we should focus on finding or creating a transplantation site that, more than the unmodified peritoneal cavity, permits for close contact between the blood and the encapsulated islet tissue. PMID- 10389843 TI - Cow's milk formula feeding induces primary immunization to insulin in infants at genetic risk for type 1 diabetes. AB - Insulin autoantibodies (IAAs) often appear as the first sign of islet cell autoimmunity in prediabetic children. Because cow's milk contains bovine insulin, we followed the development of insulin-binding antibodies in children fed with cow's milk formula. Bovine insulin- and human insulin-binding antibodies by enzyme immunoassay and IAA by radioimmunoassay were analyzed in 200 infants carrying HLA-DQB1*0302 but no protective alleles who participated in a Finnish population-based birth-cohort study. Based on the prospectively registered information, the first 100 infants enrolled in the study who were exposed to cow's milk formula before age 12 weeks and the first 100 infants enrolled in the study who were exclusively breast-fed for longer than their first 12 weeks of life were selected for the present study. Also, 11 children from the birth cohort who developed at least two diabetes-associated autoantibodies, 98 children with newly diagnosed type 1 diabetes, and 92 healthy children were studied. We found that the amount of IgG-antibodies binding to bovine insulin was higher at age 3 months in infants who were exposed to cow's milk formula than in infants who were exclusively breast-fed at that age (median 0.521 vs. 0.190; P < 0.0001). The antibodies binding to bovine insulin cross-reacted with human insulin. None of these infants tested positive for IAA. The levels of bovine insulin-binding antibodies declined in both groups at ages 12 and 18 months, whereas in the 11 children with at least two diabetes-associated autoantibodies the levels increased during the follow-up period (P < 0.0001). IgG antibodies correlated with IgG2 antibodies binding to bovine insulin (r = 0.43, P = 0.004) and IAA (r = 0.27, P = 0.02) in diabetic children, but not in healthy children. Cow's milk feeding is an environmental trigger of immunity to insulin in infancy that may explain the epidemiological link between the risk of type 1 diabetes and early exposure to cow's milk formulas. This immune response to insulin may later be diverted into autoaggressive immunity against beta-cells in some individuals, as indicated by our findings in children with diabetes-associated autoantibodies. PMID- 10389844 TI - Long-term elevation of free fatty acids leads to delayed processing of proinsulin and prohormone convertases 2 and 3 in the pancreatic beta-cell line MIN6. AB - To explore the role of chronically elevated free fatty acids (FFAs) in the pathogenesis of the hyperproinsulinemia of type 2 diabetes, we have investigated the effect of FFAs on proinsulin processing and prohormone convertases PC2 and PC1/PC3 in MIN6 cells cultured in Dulbecco's modified Eagle's medium with or without 0.5 mmol/l FFA mixture (palmitic acid:oleic acid = 1:2). After 7 days of culture, the percent of proinsulin in FFA-exposed cells was increased (25.9 +/ 0.3% intracellular and 75.4 +/- 1.2% in medium vs. 13.5 +/-0.2 and 56.2 +/- 4.1%, respectively, in control cells). The biosynthesis and secretion of proinsulin and insulin were analyzed by comparing the incorporation of [3H]Leu and [35S]Met. In pulse-chase studies, proinsulin-to-insulin conversion was inhibited, and proinsulin in the medium was increased by 50% after 3 h of chase, while insulin secretion was decreased by 50% after FFA exposure. Levels of cellular PC2 and PC3 analyzed by Western blotting were decreased by 23 and 15%, respectively. However, PC2, PC3, proinsulin, and 7B2 mRNA levels were not altered by FFA exposure. To test for an effect on the biosynthesis of PC2, PC3, proinsulin, and 7B2, a protein required for PC2 activation, MIN6 cells were labeled with [35S]Met for 10 15 min, followed by a prolonged chase. Most proPC2 was converted after 6 h of chase in control cells, but conversion was incomplete even after 6 h of chase in FFA-exposed MIN6 cells. Media from chase incubations showed that FFA-exposed cells secreted more proPC2 than controls. Similar inhibitory effects were noted on the processing of proPC3, proinsulin, and 7B2. In conclusion, prolonged exposure of beta-cells to FFAs may affect the biosynthesis and posttranslational processing of proinsulin, PC2, PC3, and 7B2, and thereby contribute to the hyperproinsulinemia of type 2 diabetes. The mechanism of inhibition of secretory granule processing by FFAs may be through changes in Ca2+ concentration, the pH in the secretory granules, and/or other factors that may influence the activation and function of the convertases. PMID- 10389845 TI - Pancreatic beta-cell-to-beta-cell interactions are required for integrated responses to nutrient stimuli: enhanced Ca2+ and insulin secretory responses of MIN6 pseudoislets. AB - The effect of cell-to-cell contact on Ca2+ influx and secretory responses in the beta-cell line MIN6 was studied using MIN6 pseudoislets, which are three dimensional islet-like cell aggregates that develop when MIN6 cells are cultured for 6-8 days on gelatin. The formation of pseudoislets is dependent on the Ca2+ dependent adhesion molecule E-cadherin (E-CAD), since the process can be inhibited by incubation in the absence of Ca2+ or in the presence of an anti-E CAD antibody. Glucose and alpha-ketoisocaproic acid (KIC) evoked a Ca2+ influx in only a small fraction of the MIN6 monolayer cells, whereas >80% of cell groups within the pseudoislets responded to both nutrients. In contrast, changes in the intracellular free Ca2+ concentration ([Ca2+]i) were observed in all or most monolayer cells or pseudoislet cell groups in response to physical or pharmacological depolarizing stimuli. No significant increase in insulin release was observed from MIN6 monolayer cells in response to nutrient or nonnutrient insulin secretagogues. Conversely, pseudoislets were found to respond significantly to both nutrients and nonnutrients. These results suggest that close cell-to-cell contact improves the functional responsiveness of MIN6 cells and that pseudoislets may therefore serve as a useful research model in the study of beta-cell function. PMID- 10389846 TI - Enhancing effects of long-term elevated glucose and palmitate on stored and secreted proinsulin-to-insulin ratios in human pancreatic islets. AB - Relative hypersecretion of proinsulin is a feature of type 2 diabetes. We investigated to what extent this feature can be induced in human pancreatic islets by elevated glucose or fatty acids, two major abnormalities of the diabetic state. A 48-h culture period with 27 mmol/l glucose increased the intraislet proinsulin-to-insulin (PI/I) ratio 5.0-fold, owing to preferential decrease of insulin. The PI/I ratio in culture medium was enhanced 1.9-fold versus islets cultured with 5.5 mmol/l glucose. This effect of elevated glucose persisted after normalization of glucose levels: during 60-min postculture incubations at a basal glucose concentration (3.3 mmol/l), the PI/I ratio of secretion increased 4.9-fold. The ratio was also increased (14-fold) after renewed postculture stimulation with 16.7 mmol/l glucose. Diazoxide was added to culture medium to block glucose-induced insulin secretion and thus investigate the importance of overstimulation. In cultures at 27 mmol/l glucose, the presence of diazoxide decreased the PI/I ratio of islet contents by 76%, the accumulated secretion to culture medium by 70%, and the release at 3.3 or 16.7 mmol/l glucose during postculture incubations by 85 and 86%, respectively. None of these PI/I decreasing effects of diazoxide were reproduced during or after coculture with 5.5 mmol/l glucose. Culture with 0.2 mmol/l palmitate and 5.5 mmol/l glucose decreased islet contents of proinsulin and insulin and increased the secreted products in culture media without affecting PI/I ratios. During postculture conditions, however, prior palmitate culture enhanced the PI/I ratio of release at 3.3 mmol/l glucose (from 2.2 +/- 0.4 to 5.4 +/- 0.9%, P < 0.05). Culture with palmitate together with 27 mmol/l glucose decreased islet contents of proinsulin and insulin and further enhanced intraislet PI/I ratios (from 9.3 +/- 1.1 to 13.4 +/- 2.5%, P < 0.05). However, palmitate failed to affect PI/I ratios in culture medium. In contrast, in postculture incubations at 3.3 mmol/l glucose, prior palmitate culture further elevated the PI/I ratio of secretion (from 10.8 +/- 1.2 after previous 27 mmol/l glucose alone to 13.9 +/- 2.8% after palmitate and glucose, P < 0.05). We conclude that 1) long-term exposure of human islets to elevated glucose leads to preferential secretion of proinsulin, and this effect persists also after glucose normalization; 2) the glucose effect appears secondary to depletion of mature insulin granules; and 3) elevated fatty acids influence PI/I ratios of secretion by mechanisms that are, in part, incongruous with an over-stimulation effect. PMID- 10389848 TI - Obesity-related phenotypes and the beta3-adrenoceptor gene variant in postmenopausal women. AB - We examined the hypothesis that postmenopausal women with the beta3-adrenoceptor gene variant (Trp64Arg) have reduced total daily energy expenditure (TEE), altered free fatty acid kinetics, and increased intra-abdominal fat. A secondary objective was to examine whether the obese state masks the effect of the variant on resting metabolic rate (RMR). There were 23 obese heterozygous women with the genetic variant (age 58 +/- 6 years; BMI 36 +/- 7 kg/m2) who were compared with 19 homozygous obese women with the normal allele (age 56 +/- 4 years; BMI 36 +/- 3 kg/m2). Daily energy expenditure was determined from doubly labeled water and indirect calorimetry, lipolysis from infusion of [1-13C]palmitate, and body fat distribution from computed tomography. No significant differences were found in TEE, RMR, energy expenditure of physical activity, the thermic effect of a meal, fat oxidation as estimated by fasting and postprandial respiratory quotients (RQs), or rate of lipolysis. Similarly, no difference was found in visceral adipose tissue and abdominal subcutaneous fat areas. When RMR was compared between obese (n = 23) and never-obese women with the Trp64Arg variant (n = 16), we found a 317 kcal/day lower RMR in never-obese women after controlling for fat mass, fat-free mass, and age (P < 0.0017). These results do not support the hypothesis that already obese women with the Trp64Arg polymorphism of the beta3 adrenergic receptor gene have lower daily energy expenditure, altered lipolysis, and increased abdominal obesity. On the other hand, the lower RMR in never-obese women suggests that the obese state may mask a moderate effect of the Trp64Arg variant on energy expenditure. Although these results need to be confirmed in other populations, the obese state may have been a confounding factor in previous studies of the beta3-adrenoceptor Trp64Arg variant and energy expenditure. PMID- 10389847 TI - A novel N-aryl tyrosine activator of peroxisome proliferator-activated receptor gamma reverses the diabetic phenotype of the Zucker diabetic fatty rat. AB - The discovery that peroxisome proliferator-activated receptor (PPAR)-gamma was the molecular target of the thiazolidinedione class of antidiabetic agents suggested a key role for PPAR-gamma in the regulation of carbohydrate and lipid metabolism. Through the use of high-throughput biochemical assays, GW1929, a novel N-aryl tyrosine activator of human PPAR-gamma, was identified. Chronic oral administration of GW1929 or troglitazone to Zucker diabetic fatty (ZDF) rats resulted in dose-dependent decreases in daily glucose, free fatty acid, and triglyceride exposure compared with pretreatment values, as well as significant decreases in glycosylated hemoglobin. Whole body insulin sensitivity, as determined by the euglycemic-hyperinsulinemic clamp technique, was significantly increased in treated animals. Comparison of the magnitude of glucose lowering as a function of serum drug concentrations showed that GW1929 was 2 orders of magnitude more potent than troglitazone in vivo. These data were consistent with the relative in vitro potencies of GW1929 and troglitazone. Isolated perfused pancreas studies performed at the end of the study confirmed that pancreata from vehicle-treated rats showed no increase in insulin secretion in response to a step change in glucose from 3 to 10 mmol/l. In contrast, pancreata from animals treated with GW1929 showed a first- and second-phase insulin secretion pattern. Consistent with the functional data from the perfusion experiments, animals treated with the PPAR-gamma agonist had more normal islet architecture with preserved insulin staining compared with vehicle-treated ZDF rats. This is the first demonstration of in vivo efficacy of a novel nonthiazolidinedione identified as a high-affinity ligand for human PPAR-gamma. The increased potency of GW1929 compared with troglitazone both in vitro and in vivo may translate into improved clinical efficacy when used as monotherapy in type 2 diabetic patients. In addition, the significant improvement in daily meal tolerance may impact cardiovascular risk factor management in these patients. PMID- 10389849 TI - Hypoglycemia per se stimulates sympathetic neural as well as adrenomedullary activity, but, unlike the adrenomedullary response, the forearm sympathetic neural response is not reduced after recent hypoglycemia. AB - We tested the hypotheses that 1) hypoglycemia per se stimulates the sympathetic neural as well as the adrenomedullary component of the sympathochromaffin system, and 2) sympathetic neural responses to hypoglycemia, like adrenomedullary responses, are reduced after recent hypoglycemia. To this end, we studied 10 healthy young adults on 2 consecutive days on two separate occasions, on one occasion with euglycemia (5.0 mmol/l) and on the other occasion with hypoglycemia (2.8 mmol/l) from 1000 to 1200 and 1400 to 1600 on day 1 of each occasion. On day 2 of each occasion, plasma epinephrine and norepinephrine (NE) concentrations and rates of systemic NE spillover (SNESO) and forearm NE spillover (FNESO) were measured during hyperinsulinemic (12.0 pmol x kg(-1) x min(-1)) euglycemia (5.0 mmol/l) and hypoglycemia (2.8 mmol/l). Compared with values during euglycemia, plasma epinephrine and NE and rates of SNESO and FNESO all increased during hypoglycemia (P < 0.01). After day 1 hypoglycemia, there were reductions during hypoglycemia on day 2 in plasma epinephrine (2,050 +/- 500 vs. 2,960 +/- 400 pmol/l; P < 0.02), plasma NE (1.35 +/- 0.16 vs. 1.92 +/- 0.20 nmol/l; P < 0.01), and SNESO rates (5.13 +/- 0.84 vs. 6.87 +/- 0.81 nmol/min; P < 0.02). However, FNESO rates were unaltered (1.16 +/- 0.25 vs. 1.27 +/- 0.17 pmol x min(-1) x 100 ml tissue(-1). Thus we conclude that 1) hypoglycemia per se stimulates both the sympathetic neural and adrenomedullary components of the sympathochromaffin system and 2) adrenomedullary, but not forearm sympathetic neural, responses to hypoglycemia are reduced after recent hypoglycemia. The extent to which the lower plasma NE levels and reduced SNESO responses to hypoglycemia after day 1 hypoglycemia reflect reduced NE release from the adrenal medullae, sympathetic nerves other than those in the forearm, or both cannot be determined from these data. PMID- 10389850 TI - New concept for long-acting insulin: spontaneous conversion of an inactive modified insulin to the active hormone in circulation: 9-fluorenylmethoxycarbonyl derivative of insulin. AB - Insulin is a short-lived species in the circulatory system. After binding to its receptor sites and transmission of its biological signals, bound insulin undergoes receptor-mediated endocytosis and consequent degradation. An inactive insulin derivative that is not recognized by the receptor has a longer circulation life, but obviously is biologically impotent. (Fmoc)2 insulin is an insulin derivative purified through high-performance liquid chromatography in which two 9-fluorenylmethoxycarbonyl (Fmoc) moieties are covalently linked to the (alpha-amino group of phenylalanine B1 and the epsilon-amino group of lysine B29. It has 1-2% of the biological potency and receptor binding capacity of the native hormone. After incubation, (Fmoc)2 insulin undergoes a time-dependent spontaneous conversion to fully active insulin in aqueous solution at 37 degrees C and a pH range of 7-8.5. At pH 7.4, the conversion proceeds slowly (t1/2 = 12 +/- 1 days) and biological activity is generated gradually. A single subcutaneous administration of (Fmoc)2 insulin to streptozocin-treated diabetic rats normalized their blood glucose levels and maintained the animals in an anabolic state over 2-3 days. A broad shallow peak of immunoreactive insulin was found to persist in circulation over this period. To confirm further that the long-acting effect of (Fmoc)2 insulin proceeds via slow release in the blood circulation itself, we administered native insulin, NPH insulin, or the (Fmoc)2 derivative intraperitoneally. The rats recovered from hypoglycemia at t1/2 = 8.0 +/- 0.3 and 10 +/- 0.4 h after administration of native and NPH insulin, respectively. In contrast, (Fmoc)2 insulin was active for a significantly longer time, with an extended onset of t1/2 = 26 +/- 1h, and a glucose-lowering effect even 40 h after administration. (Fmoc)2 insulin was also found to be more resistant to proteolysis. Finally, we found that (Fmoc)2 insulin does not induce antigenic effects. In summary, we present here a new concept for prolonging the half-life of insulin in the circulatory system, in which receptor-mediated endocytosis and degradation is delayed and accompanied by a time-dependent generation of basal insulin. PMID- 10389851 TI - Effects of glucose intolerance on myocardial function and collagen-linked glycation. AB - In experimental diabetes, diastolic dysfunction of the left ventricle has been associated with collagen-linked glycation. To determine whether less severe hyperglycemia may have similar effects, we gave alloxan to mongrel dogs (group 2) to induce impaired glucose tolerance (IGT) for comparison with normal subjects (group 1). After 6 months, hemodynamic studies were performed in the anesthetized animals. Basal heart rate, aortic pressure, and ejection fraction were comparable in the two groups, but calculated chamber stiffness was increased in group 2, associated with a reduced end diastolic volume and increased pressure. During infusion of dextran, the volume and pressure responses were similarly abnormal in group 2. In the myocardium, the collagen concentration rose with an increased interstitial distribution histologically. To assess glycation, collagen was extracted, digested with collagenase, and measured for fluorescence. Advanced glycation end products were increased in group 2 to 10.6 +/- 1.6 vs. 6.9 +/- 0.7 fluorescent units (FU)/mg collagen in group 1 (P < 0.01). To assess whether this could be pharmacologically prevented, we administered enalapril to inhibit ACE during the 6 months of glucose intolerance to group 3. This resulted in normal glycation and significant reduction in chamber stiffness increment. We gave group 4 animals aminoguanidine daily for 6 months, which prevented abnormal collagen glycation and chamber stiffness. Thus, in animals with IGT, collagen-linked glycosylation appeared to be a major factor affecting diastolic function and was shown to be amenable to pharmacological intervention. PMID- 10389852 TI - The thiazolidinedione rosiglitazone (BRL-49653) lowers blood pressure and protects against impairment of endothelial function in Zucker fatty rats. AB - Human obesity is associated with insulin resistance, hyperinsulinemia, and a predisposition to hypertension and vascular disease, the origin of which may lie in impairment of endothelial function. We tested the effects of the thiazolidinedione rosiglitazone on blood pressure and endothelial function in insulin-resistant fatty Zucker rats, which display hypertension and abnormal endothelial cell function. We studied fatty Zucker rats given rosiglitazone maleate (50 micromol/kg diet; n = 8) for 9-12 weeks (treated fatty), untreated fatty rats (n = 8), and lean rats (n = 8) given diet alone. At the end of the study, systolic blood pressure was significantly higher in untreated fatty (147 +/- 5 mmHg) than in lean rats (125 +/- 2 mmHg; P < 0.05), but rosiglitazone treatment prevented the development of hypertension in fatty rats (123 +/- 1 mmHg). Fasting hyperinsulinemia in untreated fatty rats (28.7 +/- 6.0 ng/ml) was significantly lowered by rosiglitazone (7.0 +/- 1.4 ng/ml; P < 0.05 vs. untreated fatty), but remained significantly higher than the levels seen in lean rats (1.5 +/- 0.4 ng/ml; P < 0.01). Mesenteric arteries were studied in a myograph. Maximal acetylcholine chloride (1.1 micromol/l)-induced relaxation of norepinephrine hydrochloride (NE)-induced constriction was impaired in untreated fatty (62.4 +/- 3.4%) vs. lean (74.3 +/- 3.5%; P = 0.01) rats; this defect was partially prevented by rosiglitazone (66.5 +/- 3.0%; P = 0.01 vs. untreated fatty). Insulin (50 mU/l) significantly attenuated the contractile response to NE in lean rats (14.7 +/- 3.3%; P = 0.02); this vasodilator effect of insulin was absent in untreated fatty rats at concentrations of 50-5,000 mU/l, but was partially restored by rosiglitazone (9.7 +/- 2.5% attenuation; P = 0.02 vs. no insulin). Thus, rosiglitazone prevents the development of hypertension and partially protects against impaired endothelial function associated with insulin resistance. These latter effects may contribute to the drug's antihypertensive properties. PMID- 10389853 TI - Antibodies to oxidized LDL predict coronary artery disease in type 1 diabetes: a nested case-control study from the Pittsburgh Epidemiology of Diabetes Complications Study. AB - The pathogenesis of excess cardiovascular risk in type 1 diabetes is unclear. LDL cholesterol is only weakly predictive, and its concentration is often normal in type 1 diabetes. We therefore examined whether markers of LDL oxidation such as antibodies to oxidized LDL (Ab-OxLDL) and LDL-containing immune complexes, rather than LDL concentration, were predictive of coronary artery disease (CAD) in type 1 diabetes. This nested case-control study from an epidemiologic cohort study included 49 incident cases of myocardial infarction (MI), angina, or CAD death and 49 age-, sex-, and duration-matched control subjects. Ab-OxLDL was measured by enzyme immunoassay and the apolipoprotein B (ApoB) content of immune complexes (ApoB-IC) precipitated by polyethylene glycol by immunoelectrophoresis in baseline stored samples. Ab-OxLDL was inversely, and ApoB-IC directly, related to subsequent CAD. In multivariate analyses, Ab-OxLDL remained a significant independent predictor along with previously recognized predictors, hypertension and Beck depression score. In conclusion, oxidation of LDL and the immune response it elicits may play a role in predicting the development of CAD in type 1 diabetes and explain at least some of the enhanced CAD risk in type I diabetes. PMID- 10389854 TI - Functional characterization of the MODY1 gene mutations HNF4(R127W), HNF4(V255M), and HNF4(E276Q). AB - Genetic studies have shown that mutations in the gene encoding hepatocyte nuclear factor (HNF)-4alpha, a member of the steroid/thyroid hormone receptor superfamily, give rise to early-onset type 2 diabetes (MODY1). The functional properties of mutant HNF-4alpha proteins and the molecular mechanisms by which they impair insulin secretion are largely unknown. In the present study, we have investigated transcriptional activation, DNA binding properties, and protein dimerization activity of three HNF-4alpha missense mutations--HNF4(R127W), HNF4(V255M), and HNF4(E276Q)--that have been associated with type 2 diabetes. We demonstrate that HNF4(E276Q) has lost its ability to bind to HNF-4 consensus binding sites and activate transcription. HNF4(E276Q) had no effect on the functional activity of wild-type HNF-4alpha in the pancreatic beta-cell line HIT T15, but it exhibited weak dominant-negative activity in other cell types. Analysis of HNF4(E276Q) protein showed that it exists in two forms: a full length 54-kDa protein and a 40-kDa COOH-terminal protein lacking the NH2-terminal transactivation domain and the DNA binding domain. Immunoprecipitation experiments indicate that this truncated protein can bind to wild-type HNF-4alpha and may be responsible for the weak dominant-negative effects seen in these cells. In addition, we show that the transcriptional transactivation of HNF4(R127W) and HNF4(V255M) is indistinguishable from that of wild-type HNF 4alpha, suggesting that they are sequence polymorphisms. Our results demonstrate that HNF4(E276Q) is a loss-of-function mutation and that it identifies glutamic acid 276 in alpha-helix 8 of the ligand-binding domain of HNF-4alpha protein as a critical residue for DNA binding, transcriptional activation, and protein stability in vivo. PMID- 10389855 TI - Pro12Ala substitution in the peroxisome proliferator-activated receptor-gamma2 is not associated with type 2 diabetes. AB - Peroxisome proliferator-activated receptor (PPAR)-gamma is a major regulator of adipogenesis and insulin sensitivity. The PPAR-gamma gene generates two isoforms through alternative splicing, PPAR-gamma1 and -gamma2, the latter having an additional stretch of 28 amino acids at its NH2-terminus in the ligand independent activation domain. This extension renders PPAR-gamma2 more sensitive to insulin action. Since there is a Pro12Ala substitution in this domain, we tested whether it is related to type 2 diabetes or insulin resistance. Therefore, 131 type 2 diabetic patients and 312 normoglycemic control subjects were screened for the presence of the mutation and for major clinical and metabolic features. The frequency of the mutation did not differ significantly between diabetic patients and control subjects. BMI, insulin, and other metabolic and anthropometric variables were also not associated with the mutation. Although the study was carried out on a sufficiently large sample, the conclusions do not support a major role for the Pro12Ala substitution of the PPAR-gamma gene in the etiology of type 2 diabetes. PMID- 10389856 TI - The 3'-untranslated region polymorphism of the gene for skeletal muscle-specific glycogen-targeting subunit of protein phosphatase 1 in the type 2 diabetic Japanese population. AB - A newly identified 3'-untranslated region (UTR) polymorphism of the gene for skeletal muscle-specific glycogen-targeting subunit of protein phosphatase 1 (PPP1R3) was associated with insulin resistance and type 2 diabetes in Pima Indians (Xia J, Scherers W, Cohen PTW, Majer M, Xi T, Norman RA, Knowler WC, Bogardus C, Prochazka M: A common variant in PP1R3 associated with insulin resistance and type 2 diabetes. Diabetes 47:1519-1524, 1998). Thus, we investigated the frequency of polymorphism of the adenine- and thymine-rich element (ARE-1 and its variant ARE-2) in 426 Japanese type 2 diabetic and 380 nondiabetic subjects using a polymerase chain reaction (PCR)-restriction enzyme fragment length polymorphism (RFLP) method. The allele frequency of the ARE-2 variant in diabetic subjects was higher than that in nondiabetic subjects (0.34 vs. 0.29; P < 0.05), even though its frequency in Japanese subjects was lower (P < 0.001) than the reported value in Pima Indians (0.56). An aspartate polymorphism at codon 905 was 100% coupled to the ARE-2 allele, and its allele frequency was higher also in diabetic subjects. Although a serine substitution at codon 883 was partially linked with the ARE-2 allele, there was no difference between diabetic and nondiabetic subjects. These results indicate that the frequency of polymorphism of the PPP1R3 gene (ARE-2 and Asp905) is different between two ethnic groups and is increased in Japanese people with type 2 diabetes, suggesting that these variants may be a possible marker for searching for diabetogenic genes. PMID- 10389857 TI - Long-term survival and function of intrahepatic islet allografts in baboons treated with humanized anti-CD154. AB - Clinical islet cell transplantation has resulted in insulin independence in a limited number of cases. Rejection, recurrence of autoimmunity, and impairment of normal islet function by conventional immunosuppressive drugs, e.g., steroids, tacrolimus, and cyclosporin A, may all contribute to islet allograft loss. Furthermore, intraportal infusion of allogeneic islets results in the activation of intrahepatic macrophages and endothelial cells, followed by production of proinflammatory mediators that can contribute to islet primary nonfunction. We reasoned that the beneficial effects of anti-CD154 treatment on autoimmunity, alloreactivity, and proinflammatory events mediated by macrophages and endothelial cells made it an ideal agent for the prevention of islet allograft failure. In this study, a nonhuman primate model (Papio hamadryas) was used to assess the effect of humanized anti-CD154 (hu5c8) on allogeneic islet engraftment and function. Nonimmunosuppressed and tacrolimus-treated recipients were insulin independent posttransplant, but rejected their islet allografts in 8 days. Engraftment and insulin independence were achieved in seven of seven baboon recipients of anti-CD154 induction therapy administered on days -1, 3, and 10 relative to the islet transplant. Three of three baboons treated with 20 mg/kg anti-CD154 induction therapy experienced delayed rejection episodes, first detected by elevations in postprandial blood glucose levels, on postoperative day (POD) 31 for one and on POD 58 for the other two. Re-treatment with three doses of anti-CD154 resulted in reversal of rejection in all three animals and in a return to normoglycemia and insulin independence in two of three baboons. It was possible to reverse multiple episodes of rejection with this approach. A loss of functional islet mass, as detected by reduced first-phase insulin release in response to intravenous glucose tolerance testing, was observed after each episode of rejection. One of two baboons treated with 10 mg/kg induction therapy became insulin independent post-transplant but rejected the islet graft on POD 10; the other animal experienced a reversible rejection episode on POD 58 and remained insulin independent and normoglycemic until POD 264. Two additional baboon recipients of allogeneic islets and donor bone marrow (infused on PODs 5 and 11) were treated with induction therapy (PODs -1, 3, 10), followed by initiation of monthly maintenance therapy (for a period of 6 months) on POD 28. Rejection-free graft survival and insulin independence was maintained for 114 and 238 days, with preservation of functional islet mass observed in the absence of rejection. Prevention and reversal of rejection, in the absence of the deleterious effects associated with the use of conventional immunosuppressive drugs, make anti-CD154 a unique agent for further study in islet cell transplantation. PMID- 10389858 TI - Overexpression of uncoupling protein 2 inhibits glucose-stimulated insulin secretion from rat islets. AB - Uncoupling protein 2 (UCP-2) mRNA expression has been shown to be altered by metabolic conditions such as obesity in humans, but its functional significance is unknown. The expression of UCP-2 mRNA and protein in normal rat islets was established by reverse transcriptase-polymerase chain reaction and immunocytochemistry in pancreatic islets and tissue, respectively. Intense immunostaining of UCP-2 correlated with insulin-positive ,-cells. Overexpression of UCP-2 in normal rat islets was accomplished by infection with an adenovirus (AdEGI-UCP-2) containing the full-length human UCP-2 coding sequence. Induction of the AdEGI-UCP-2 gene resulted in severe blunting of glucose-stimulated insulin secretion (GSIS) without affecting islet insulin content or the ability of the calcium ionophore A23187 to increase insulin secretion from AdEGI-UCP-2 expressing islets. Therefore, UCP-2 overexpression affects signal transduction proximal to Ca2+-mediated steps, including exocytosis. Insulin secretion from single beta-cells to 16.5 mmol/l glucose examined by reverse hemolytic plaque assay was nearly ablated if UCP-2 was overexpressed. Thus, a direct, causal relationship between overexpression of UCP-2 and inhibition of GSIS in normal islets has been established. These data suggest that increased expression of UCP 2 has the potential to cause the lack of a glucose effect on insulin secretion in type 2 diabetes. PMID- 10389859 TI - Leptin restores euglycemia and normalizes glucose turnover in insulin-deficient diabetes in the rat. AB - Leptin has been shown to improve insulin sensitivity and glucose metabolism in normoinsulinemic healthy or obese rodents. It has not been determined whether leptin may act independently of insulin in regulating energy metabolism in vivo. The present study was designed to examine the effects of leptin treatment alone on glucose metabolism in insulin-deficient streptozotocin (STZ)-induced diabetic rats. Four groups of STZ-induced diabetic rats were studied: 1) rats treated with recombinant methionine murine leptin subcutaneous infusion with osmotic pumps for 12-14 days (LEP; 4 mg x kg(-1) x day(-1), n = 10); 2) control rats infused with vehicle (phosphate-buffered saline) for 12-14 days (VEH; n = 10); 3) pair-fed control rats given a daily food ration matching that of LEP rats for 12-14 days (PF; n = 8); and 4) rats treated with subcutaneous phloridzin for 4 days (PLZ; 0.4 g/kg twice daily, n = 10). Phloridzin treatment normalizes blood glucose without insulin and was used as a control for the effect of leptin in correcting hyperglycemia. All animals were then studied with a hyperinsulinemic-euglycemic clamp (6 mU x kg(-1) x min(-1). Our study demonstrates that leptin treatment in the insulin-deficient diabetic rats restored euglycemia, minimized body weight loss due to food restriction, substantially improved glucose metabolic rates during the postabsorptive state, and restored insulin sensitivities at the levels of the liver and the peripheral tissues during the glucose clamp. The effects on glucose turnover are largely independent of food restriction and changes in blood glucose concentration, as evidenced by the minimal improvement of insulin action and glucose turnover parameters in the PF and PLZ groups. Our results suggest that the antidiabetic effects of leptin are achieved through both an insulin independent and an insulin-sensitizing mechanism. PMID- 10389860 TI - Value of laparoscopic staging for upper gastrointestinal malignancies. PMID- 10389861 TI - Are we ready to certify surgical oncologists? PMID- 10389862 TI - ret/PTC expression may be associated with local invasion of thyroid papillary carcinoma. AB - BACKGROUND AND OBJECTIVES: The exact role of ret/PTC in the development of papillary carcinoma remains unclear. Expression of the ret/PTC oncogene was examined immunohistochemically to address its role in the progression of thyroid carcinomas. METHODS: Paraffin-embedded samples from 34 clinically evident thyroid papillary carcinomas and 19 occult papillary carcinomas were analyzed using an antibody raised against the ret tyrosine kinase domain. RESULTS: Expression of ret/PTC was demonstrated in 6/19 (32%) occult carcinomas. The frequency of expression of ret/PTC in clinically evident carcinomas in 16/34 (47%) was significantly higher than in normal tissues (0%) and follicular adenomas (1/14, 7%, P < 0.01).ret/PTC expression was observed more frequently in the peripheral areas of clinically evident carcinomas (P < 0.01). Although there was no correlation of ret/PTC expression with tumor size, lymph node metastasis, or distant metastasis, the incidence of ret/PTC expression in tumors with extrathyroidal invasion (13/19, 68%) was significantly higher than those without extrathyroidal invasion (3/15, 20%, P < 0.01). Local invasion was found in none of the occult carcinomas. The frequency of expression in occult carcinomas was significantly lower than in clinically evident carcinomas with extrathyroidal invasion (P < 0.05). CONCLUSIONS: The ret/PTC oncogene may be involved in the local invasion of thyroid papillary carcinomas. PMID- 10389863 TI - Rapid development of hepatic metastasis with high incidence following orthotopic transplantation of murine colon 38 carcinoma as intact tissue in syngeneic C57BL/6 mice. AB - BACKGROUND AND OBJECTIVES: Orthotopic transplantation of human colon tumors was a useful method for producing hepatic metastasis in mice. In many cases, however, it took about 3 months for evaluation. We examined an in vivo model of hepatic metastasis for only 4 weeks by conducting orthotopic transplantation of murine Colon 38 tumor using intact tissue in syngeneic mice and determined the efficacy of chemotherapeutic agents against hepatic metastasis. METHODS: Twenty milligrams of tumor tissues were prepared from subcutaneously (s.c.) growing Colon 38 tumor and orthotopically transplanted on the cecum in C57BL/6 mice. Mice were autopsied about 4 weeks after transplantation. Metastases to various organs were detected macroscopically or histochemically and tumor invasion into the cecum was observed histochemically. In experimental chemotherapy, mice bearing orthotopically transplanted Colon 38 tumor were separated into three equal groups and were either treated with fluorouracil or cisplatin (CDDP), or untreated. Four weeks after transplantation, activities of both agents against local tumor growth and hepatic metastasis were evaluated. RESULTS: Macroscopic metastases to various organs including the liver, the lung, and the peritoneum were developed during days 28 to 32 after inoculation. The frequency of hepatic metastasis was 96% (N = 23). Histological examination indicated that the local tumor invaded various layers of the cecum and metastasized to the liver and lung hematogenously. In experimental chemotherapy with fluorouracil and CDDP, only fluorouracil decreased the incidence of mice with hepatic metastasis (2/8 cases), compared with vehicle treatment (7/8 cases) and the number of metastatic nodules in the liver (P = 0.016), although the inhibition against local growth of CDDP in T/C [45%; mean tumor weight of the test group (T) compared with that of the control group (C)] was similar to that of fluorouracil (53%). CONCLUSIONS: This model, with its rapid development of hepatic metastasis in high frequency, should be useful as a screening assay to find anti-metastatic agents for colorectal carcinoma. PMID- 10389864 TI - Irradiation-induced up-regulation of Fas in esophageal squamous cell carcinoma is not accompanied by Fas ligand-mediated apoptosis. AB - BACKGROUND AND OBJECTIVES: Fas (APO-1) induces apoptosis after binding Fas ligand (FasL). Evidence suggests that tumors may use this interaction to evade the host immune response. Fas/FasL expression has not been reported in esophageal cancer. We hypothesized that Fas expression would render esophageal cancer cells susceptible to Fas ligation and that irradiation of the cells would increase Fas expression. METHODS: Two human esophageal squamous cell carcinoma lines, KYSE 150, which has a wild-type (wt) p53 gene, and 410 (mutated p53), were irradiated. Reverse-transcriptase polymerase chain reaction was used to detect Fas and FasL expression. Fas protein was quantitated by enzyme-linked immunosorbent assay and its presence further confirmed by Western analysis. FasL was detected by Western analysis. Cells were treated with Fas monoclonal antibody (maximum 0.05 microg/ml)+/-cycloheximide, and viability was assessed by 3-(4,5-dimethylthiazol 2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cells were also transduced with FasL cDNA and then quantified. RESULTS: Both lines expressed Fas and FasL, but only the KYSE 150 cell line displayed an increase in Fas following irradiation. No alteration in cell growth was detected for Fas antibody- or FasL transduced groups versus controls. CONCLUSIONS: We have demonstrated Fas and FasL expression in esophageal tumor lines. We have also shown that Fas levels are significantly increased in response to irradiation in a wt p53 line. However, cells were resistant to treatment with Fas antibody or following transduction with FasL, suggesting that these tumor cells may use Fas/FasL expression to evade the host immune response. PMID- 10389865 TI - Laparoscopic-assisted resection of right-sided colonic carcinoma: a case-control study. AB - BACKGROUND AND OBJECTIVES: Laparoscopic-assisted resection of colorectal carcinoma is technically feasible. Whether it is beneficial to patients is uncertain. This study reviewed the results of laparoscopic-assisted resection in patients with right-sided colonic adenocarcinoma. METHODS: We attempted laparoscopic-assisted right to extended right hemicolectomy in 28 patients with right-sided colonic carcinoma (study group). The results were compared with 56 matched patients who underwent conventional open resection in the same period (comparative group). RESULTS: The median follow-up times for the study and comparative groups were 21.4 and 23.5 months, respectively. The operating time was significantly longer (t-test, P < 0.001), whereas the time to resuming normal diet (Mann-Whitney U-test, P < 0.001) and the duration of hospital stay (Mann Whitney U-test, P = 0.002) were significantly less in the study than in the comparative group. The oncological clearance, in terms of the number of lymph nodes removed and the resection margins, the complication rate, the disease-free rate, and the survival rate were comparable in the two groups. CONCLUSIONS: We conclude that laparoscopic-assisted resection of right-sided colonic adenocarcinoma has the advantage over open surgery of allowing earlier recovery. However, this is at the expense of a longer operating time. PMID- 10389866 TI - Prognosis of a series of 763 consecutive node-negative invasive breast cancer patients without adjuvant therapy: analysis of clinicopathological prognostic factor. AB - BACKGROUND AND OBJECTIVES: The objectives of this study were to confirm the favorable outcome of Japanese invasive breast cancer patients without lymph node metastasis, after treatment with surgery alone, and to evaluate clinicopathological prognostic factors in this population. METHODS: The subjects were 763 consecutive node-negative invasive breast cancer patients who underwent surgery without adjuvant therapies between 1988 and 1993 at our hospital. Disease free survival (DFS) and overall survival (OS) rates were analyzed by clinicopathological factors. RESULTS: The median age of the patients at surgery was 52 years and the median follow-up period of patients was 74 months. At 5 years, the respective DFS and OS rates of all patients were 90.8% and 93.9%. Patients with a pathological tumor size of invasive component of more than 2 cm (319 patients) had a significantly lower DFS than those with tumors measuring 2 cm or less (361 patients) (P = 0.045). Patients with positive hormone receptor status (280 patients) (estrogen and/or progesterone receptor positive) tended to have a better OS than those negative for both hormone receptors (92 patients) (P = 0.078). Meanwhile, patients with tumors of histological grade 3 (328 patients) had a much poorer prognosis than those with tumors of histological grade 1 or 2 (413 patients) (P = 0.008 for OS and P = 0.042 for DFS). The respective 5-year DFS and OS rates of patients with histological grade 3 tumors larger than 2 cm in pathological tumor size of invasive component (195 patients) were 85.5% and 87.6%, indicating that these node-negative patients form a high risk group. CONCLUSIONS: Japanese invasive breast cancer patients without lymph node metastasis tended to show a survival advantage compared with their Caucasian counterparts. Histological grade was the most useful prognostic factor in this population. PMID- 10389867 TI - Considerations during clinical operation of two commercially available cryomachines. AB - BACKGROUND AND OBJECTIVES: Advances in the technology of cryomachines in the last 10 years have led to the development of both liquid nitrogen and argon-based Joule-Thompson cryomachines. Theoretical and practical evaluation of the CMS Accuprobe and the ENDOcare CRYOcare was performed as respective examples of these technologies. METHODS: Thermal gradients were calculated about both probes for the best case scenario of probe surface temperature equaling that of the cryogen used. Also, experimental evaluation in gelatin phantoms was performed with five probe arrays. RESULTS: Theoretically, a liquid nitrogen-cooled probe provides only a slight advantage over one cooled with liquid argon. However, the experimental performance evaluation demonstrated that the CRYOcare system creates an iceball faster with steeper internal temperature gradients than the Accuprobe. Further, temperature outputs from the Accuprobe were shown to be in error, likely due to the position of the thermocouple within the probe. CONCLUSIONS: Cryomachine performance is determined more by technological innovations than by cryogen temperature. Thermocouple monitoring is urged for users of the Accuprobe. PMID- 10389869 TI - Repair after radical groin dissection. AB - The above-described repair following ilioinguinal dissection with division of the inguinal ligament is essentially a Cooper's ligament repair providing a secure, durable reconstruction. In our experience, there has not been a case of incisional hernia after radical incontinuity groin dissection using the above reconstruction. PMID- 10389868 TI - Clinical significance of serum p53 antibody detection on chemosensitivity assay in human colorectal cancer. AB - BACKGROUND AND OBJECTIVES: Alteration of the p53 gene product occurs frequently during progression of colorectal cancer. Recently, mutated p53 protein was found to induce the production of anti-p53 antibodies in the serum of patients. The purpose of this study was to evaluate the relationship between p53 status in serum and chemosensitivity in resectable colorectal cancer patients. METHODS: A total of 22 patients with primary colorectal cancer who underwent surgical treatment were examined for chemosensitivity with iable tumor samples using the Histoculture Drug Response Assay (HDRA). Serum samples of these patients for p53 antibodies were obtained before tumor resection and assayed in duplicate using an enzyme-linked immunosorbent assay kit. RESULTS: The inhibition index of 5 fluorouracil and cis-diamminedichloroplatinum (CDDP), determined by the HDRA method, in the seropositive group was significantly lower than that in the seronegative group (P < 0.01). Furthermore, significant statistical differences in chemosensitivity to 5-fluorouracil and CDDP were revealed depending on the presence of serum p53 antibodies. CONCLUSIONS: Detection of serum p53 antibodies, which reflects p53 mutations in tumor tissue, is a simpler method which correlates with chemosensitivity and may contribute to the selection of favorable chemotherapeutic strategies for colorectal cancer. PMID- 10389870 TI - Reflections and proposals for the worldwide standardization of lymphadenectomy for gastric carcinoma. PMID- 10389871 TI - Carcinoma in situ of the colorectum: SEER trends by race, gender, and total colorectal cancer. AB - BACKGROUND AND OBJECTIVES: To determine if Americans of African origin (blacks) have less access to colonoscopic polypectomy than Americans of European origin (whites), the rate of carcinoma in situ of the colorectum (CIS), a disease more similar to benign adenoma of the colorectum than invasive cancer in its symptomatology, discovery, and treatment, was determined in the United States from 1973 to 1994. The hypothesis being tested is that CIS will be far less common in blacks than in whites and that rates of CIS should be increasing in whites from 1973 to 1994. METHODS: CIS and invasive carcinoma of the colorectum incidence data were obtained from Surveillance, Epidemiology, and End Results (SEER) Public Use Files from 1973 through 1994. Rates were age adjusted and proportions determined by division of CIS rates for each subsite by total carcinoma rates, for each year, race, and gender. The colorectum was divided anatomically in this analysis at the junction of the descending and sigmoid colon. RESULTS: The relationships between male/female and black/white CIS incidence rates were broadly similar to invasive cancer rates over the 21 years of SEER, demonstrating a white male predominance for distal disease, a black male predominance for proximal disease, and a decline in incidence since 1988. CIS as a proportion of total colorectal cancer increased in all races and genders from 1973 to 1987, but then declined in all groups. CONCLUSIONS: The majority of CIS is excised by endoscopic resection. Therefore, this might be considered a surrogate population for those individuals who have colonoscopic resection of benign adenomas. It is this latter treatment that has been hypothesized to be the cause for the declining incidence of invasive colorectal cancer. However, data presented herein do not support this hypothesis. PMID- 10389872 TI - Urokinase plasminogen activator: a prognostic marker in multiple types of cancer. AB - Urokinase plasminogen activator (uPA) is a serine protease causally involved in cancer invasion and metastasis. Consistent with its role in cancer spread, uPA has been shown to be a prognostic marker in a variety of malignancies, especially breast cancer. Approximately 20 different groups have shown that high levels of uPA in breast tumor tissue predict poor outcome. As a prognostic marker in breast cancer, uPA provides information that is independent of traditionally used factors such as tumor size, tumor grade, axillary node status and estrogen receptor status. Furthermore, uPA is prognostic in node-negative patients, and a clinical trial is currently under way to assess whether uPA and its inhibitor, plasminogen activator inhibitor-1, can differentiate between the majority of node negative breast cancer patients who are cured by surgery from the minority who might benefit from adjuvant therapy. uPA is also prognostic in other malignancies, such as gastric, colorectal, esophageal, renal, endometrial, and ovarian cancers. uPA may thus be a prognostic indicator for multiple types of adenocarcinoma. PMID- 10389873 TI - Increased inspired oxygen concentration as a factor in improved brain tissue oxygenation and tissue lactate levels after severe human head injury. AB - OBJECT: Early impairment of cerebral blood flow in patients with severe head injury correlates with poor brain tissue O2 delivery and may be an important cause of ischemic brain damage. The purpose of this study was to measure cerebral tissue PO2, lactate, and glucose in patients after severe head injury to determine the effect of increased tissue O2 achieved by increasing the fraction of inspired oxygen (FiO2). METHODS: In addition to standard monitoring of intracranial pressure and cerebral perfusion pressure, the authors continuously measured brain tissue PO2, PCO2, pH, and temperature in 22 patients with severe head injury. Microdialysis was performed to analyze lactate and glucose levels. In one cohort of 12 patients, the PaO2 was increased to 441+/-88 mm Hg over a period of 6 hours by raising the FiO2 from 35+/-5% to 100% in two stages. The results were analyzed and compared with the findings in a control cohort of 12 patients who received standard respiratory therapy (mean PaO2 136.4+/-22.1 mm Hg). The mean brain PO2 levels increased in the O2-treated patients up to 359+/ 39% of the baseline level during the 6-hour FiO2 enhancement period, whereas the mean dialysate lactate levels decreased by 40% (p < 0.05). During this O2 enhancement period, glucose levels in brain tissue demonstrated a heterogeneous course. None of the monitored parameters in the control cohort showed significant variations during the entire observation period. CONCLUSIONS: Markedly elevated lactate levels in brain tissue are common after severe head injury. Increasing PaO2 to higher levels than necessary to saturate hemoglobin, as performed in the O2-treated cohort, appears to improve the O2 supply in brain tissue. During the early period after severe head injury, increased lactate levels in brain tissue were reduced by increasing FiO2. This may imply a shift to aerobic metabolism. PMID- 10389874 TI - Hemodynamic characterization of intracranial pressure plateau waves in head injury patients. AB - OBJECT: Plateau waves of intracranial pressure (ICP) are often recorded during intensive care monitoring of severely head injured patients. They are traditionally interpreted as meaningful secondary brain insults because of the dramatic decrease in cerebral perfusion pressure (CPP). The aim of this study was to investigate both the hemodynamic profile and the clinical consequences of plateau waves. METHODS: One hundred sixty head-injured patients were studied using continuous monitoring of ICP; almost 20% of these patients exhibited plateau waves. In 96 patients arterial pressure, ICP, and transcranial Doppler (TCD) blood flow velocity were studied daily for 20 minutes to 3 hours. Sixteen episodes of plateau waves in eight patients were recorded and analyzed. The dramatic increase in ICP was followed by a profound fall in CPP (by 45%). In contrast, flow velocity fell by only 20%. Autoregulation was documented to be intact both before and after plateau but was disturbed during the wave (p < 0.05). Pressure-volume compensatory reserve was always depleted before the wave. Cerebrovascular resistance decreased during the wave by 60% (p < 0.05) and TCD pulsatility increased (p < 0.05). Plateau waves did not increase the probability of an unfavorable outcome following injury. CONCLUSIONS: The authors have confirmed that the plateau waves are a hemodynamic phenomenon associated with cerebrovascular vasodilation. They are observed in patients with preserved cerebral autoregulation but reduced pressure-volume compensatory reserve. PMID- 10389875 TI - Ventricular volume following third ventriculostomy. AB - OBJECT: Ventricular size often shows no obvious change following third ventriculostomy, particularly in the early postoperative period, making postoperative evaluation difficult without expensive and often invasive testing in patients with equivocal clinical responses. The authors hypothesized that performing careful volumetric measurements would show decreases in size within the first 3 weeks after surgery. METHODS: Volumetric measurements were calculated from standard 3 x 3-mm axial computerized tomography (CT) scans obtained immediately before and 3 and 21 days after surgery. Two independent investigators measured third ventricular volume in a series of 16 patients and lateral ventricular volume in 10 of the patients undergoing stereotactically guided endoscopic third ventriculostomy for noncommunicating hydrocephalus. Fifteen patients were symptomatically improved at the time the follow-up scan was obtained. Third ventricular volume decreased in all patients by a mean of 35% (range 7.8-95.1%) and lateral ventricular volume decreased in all patients by a mean of 33% (range 4.5-80.3%). The degree of change correlated with the length of preoperative symptoms (p < 0.005). The one patient who experienced no improvement showed no decrease in third ventricular volume. In seven of 10 patients, the decrease in third ventricular volume exceeded the decrease in lateral ventricular volume. Repeated measurements indicated that the 95% confidence interval for the authors' calculations varied around the mean by 2.5% for third ventricular volume and 1.2% for lateral ventricular volume. Long-term outcome was excellent, with only one case of delayed failure. The mean follow-up duration was 12 months. CONCLUSIONS: Volumetric measurements calculated from standard CT scans will show a demonstrable decrease in ventricular volume soon after successful third ventriculostomy and can be helpful in assessing patients postoperatively. Although the third ventricle may exhibit a greater decrease, the lateral ventricular measurements are more accurate. Patients with more indolent symptoms show the smallest change. PMID- 10389876 TI - Experience with a programmable valve shunt system. AB - OBJECT: The goal of this study was to clarify the efficacy of the Codman Hakim programmable valve. Clinical data obtained in 179 patients with hydrocephalus or other intracranial fluid-accumulating diseases were analyzed. METHODS: Shunt placement operations were effective in 168 patients, approximately one half (50.6%) of whose devices required reprogramming of opening pressure postoperatively. This was a significantly larger number of shunts than the authors had thought would need reprogramming. Extremely narrowed ventricles observed on computerized tomography scans, as well as clinical symptoms related to inadequate or excessive cerebrospinal fluid drainage, improved in patients after shunt reprogramming. Shunt reprogramming frequently was necessary in patients with posthemorrhagic acute hydrocephalus; the programmable valve proved particularly beneficial for such patients. Subdural effusion and arachnoid cyst also proved to be good indications for use of the valve. Twelve patients (7%) suffered complications postoperatively. The most common complication was valve obstruction, which occurred in five patients, most of whom had brain tumors. CONCLUSIONS: The programmable valve was beneficial for the treatment of hydrocephalus and other intracranial fluid-accumulating diseases. It is important to be careful in selecting patients for treatment with the programmable valve, because complications involving the valve seem more likely in brain tumor cases. The valve proved to be poorly resistant to magnetic fields; therefore, it is essential to confirm opening pressure after every magnetic resonance imaging examination. The authors recommend that an identification system for patients be developed so that medical personnel will be aware of the presence of the valve and the previous setting of opening pressure. PMID- 10389877 TI - Low incidence of delayed intracerebral hemorrhage secondary to ventriculoperitoneal shunt insertion. AB - OBJECT: Bleeding into the brain parenchyma or ventricles is an infrequently reported complication in adults who undergo insertion of a ventriculoperitoneal (VP) shunt. The purpose of this study was to establish the incidence of delayed intracerebral hemorrhage secondary to ventricular cannulation during shunting procedures. METHODS: Over a 24-year period, in a series of 125 adult patients with hydrocephalus, postoperative computerized tomography scans were obtained in every case within 48 hours of shunt surgery performed by the same neurosurgeon. The rate of delayed intracerebral hematoma or intraventricular hemorrhage after VP shunt placement was documented by routine neuroradiological follow up to be 4%. CONCLUSIONS: In adult patients with no coagulopathy or occult vascular lesions, the rate of bleeding after VP shunt insertion may be low if the procedure is uncomplicated by multiple attempts at perforation, puncture of the choroid plexus, or improper placement of the tubing within the parenchyma of the brain. PMID- 10389878 TI - Surgery and radiotherapy compared with gamma knife radiosurgery in the treatment of solitary cerebral metastases of small diameter. AB - OBJECT: The aim of this retrospective study was to compare treatment results of surgery plus whole-brain radiation therapy (WBRT) with gamma knife radiosurgery alone as the primary treatment for solitary cerebral metastases suitable for radiosurgical treatment. METHODS: Patients who had a single circumscribed tumor that was 3.5 cm or smaller in diameter were included. Treatment results were compared between microsurgery plus WBRT (52 patients, median tumor dose 50 Gy) and radiosurgery alone (56 patients, median prescribed tumor dose 22 Gy). In case of local/distant tumor recurrence in the radiosurgery group, additional radiosurgical treatment was administered in patients with stable systemic disease. Survival time was analyzed using the Kaplan-Meier method, and prognostic factors were obtained from the Cox model. The patient groups did not differ in terms of age, gender, pretreatment Karnofsky Performance Scale (KPS) score, duration of symptoms, tumor location, histological findings, status of the primary tumor, time to metastasis, and cause of death. Patients who suffered from larger lesions underwent surgery (p < 0.01). The 1-year survival rate (median survival) was 53% (68 weeks) in the surgical group and 43% (35 weeks) in the radiosurgical group (p = 0.19). The 1-year local tumor control rates after surgery and radiosurgery were 75% and 83%, respectively (p = 0.49), and the 1 year neurological death rates in these groups were 37% and 39% (p = 0.8). Shorter overall survival time in the radiosurgery group was related to higher systemic death rates. A pretreatment KPS score of less than 70 was a predictor of unfavorable survival. Perioperative morbidity and mortality rates were 7.7% and 1.6% in the resection group, and 8.9% and 1.2% in the radiosurgery group, respectively. Four patients presented with transient radiogenic complications after radiosurgery. CONCLUSIONS: Radiosurgery alone can result in local tumor control rates as good as those for surgery plus WBRT in selected patients. Radiosurgery should not be routinely combined with radiotherapy. PMID- 10389879 TI - Long-term outcomes after meningioma radiosurgery: physician and patient perspectives. AB - OBJECT: Stereotactic radiosurgery is a primary or adjuvant management approach used to treat patients with intracranial meningiomas. The goal of radiosurgery is long-term prevention of tumor growth, maintenance of the patient's neurological function, and prevention of new neurological deficits. The object of this study is to report longer-term patient outcomes. METHODS: The authors evaluated 99 consecutive patients who underwent radiosurgery for meningioma between 1987 and 1992. Evaluation was performed using serial imaging tests, clinical evaluations, and a patient survey that was administered between 5 and 10 years after radiosurgery. Four patients underwent two radiosurgery procedures for separate meningiomas. The average tumor margin dose was 16 Gy and the median tumor volume was 4.7 ml (range 0.24-24 ml). Fifty-seven patients (57%) had undergone prior resection, of which 12 procedures were considered "total." Five patients received fractionated radiation therapy before radiosurgery. Eighty-nine patients (89%) had skull base tumors. The clinical tumor control rate (no resection required) was 93%. Sixty-one (63%) of 97 tumors became smaller, 31 (32%) remained unchanged in size, and five (5%) were enlarged. Resection was performed in seven patients (7%), six of whom had undergone prior resection. New neurological deficits developed in five patients (5%) 3 to 31 months after radiosurgery. Twenty-seven (42%) of 65 responding patients were employed at the time of radiosurgery and 20 (74%) of these remained so. Radiosurgery was believed to have been "successful" by 67 of 70 patients who completed an outcomes questionnaire 5 to 10 years later. At least one complication was described by nine patients (14%) and in four patients the complications resolved. CONCLUSIONS: Five to 10 years after radiosurgery, 96% of surveyed patients believed that radiosurgery provided a satisfactory outcome for their meningioma. Overall, 93% of patients required no other tumor surgery. Incidences of morbidity in this early experience were usually transitory and relatively mild. Radiosurgery provided long-term tumor control associated with high rates of neurological function preservation and patient satisfaction. PMID- 10389880 TI - The prophylactic use of transluminal balloon angioplasty in patients with Fisher Grade 3 subarachnoid hemorrhage: a pilot study. AB - OBJECT: Recent advances in neuroradiology have made it possible to dilate vasospastic human cerebral arteries after aneurysmal subarachnoid hemorrhage (SAH), but the time window is short and the success rate for reversal of delayed ischemic neurological deficits (DINDs) varies between 31% and 77%. In a dog model of vasospasm, transluminal balloon angioplasty (TBA) performed on Day 0 totally prevented the development of angiographically demonstrated narrowing on Day 7. Because the effect of preventive TBA in this animal model was better than any pharmacological treatment described previously for experimental vasospasm, the authors conducted a pilot trial in humans to assess the safety and efficacy of TBA performed within 3 days of SAH. METHODS: The study group consisted of 13 patients with Fisher Grade 3 SAH who had a very high probability of developing vasospasm. In all patients, regardless of the site of the ruptured aneurysm, target vessels for prophylactic TBA were as follows: the internal carotid artery, A1 segment, M1 segment, and P1 segment bilaterally; the basilar artery; and one vertebral artery. Prophylactic TBA was considered satisfactory when it could be performed in at least two of the three parts of the intracranial circulation (right and/or left carotid system and/or vertebrobasilar system), and included the aneurysm-bearing part of the circulation. Of the 13 patients, none developed a DIND or more than mild vasospasm according to transcranial Doppler ultrasonography criteria. At 3 months posttreatment eight patients had made a good recovery, two were moderately disabled, and three had died; one patient died because of a vessel rupture during TBA and two elderly individuals died of medical complications associated with poor clinical condition on admission. CONCLUSIONS: Compared with large series of patients with aneurysmal SAH reported in the literature, the results of this pilot study indicate an extremely low incidence of vasospasm and DIND after treatment with prophylactic TBA. A larger randomized study is required to determine whether prophylactic TBA is efficacious enough to justify the risks, and which vessels need to be dilated prophylactically. PMID- 10389881 TI - Clinical relevance of amygdala sclerosis in temporal lobe epilepsy. AB - OBJECT: The goal of this study was to define the incidence and clinical significance of amygdala sclerosis (AS) in patients with temporal lobe epilepsy (TLE). METHODS: Surgical specimens of the lateral amygdaloid nucleus and the hippocampus excised from 71 patients who were treated for medically intractable TLE were quantitatively evaluated using a computer-assisted image-analysis system and compared with 10 normal autopsy specimens. Densities of neurons and reactive astrocytes in the patients with TLE were correlated with clinical, neuropsychological, and depth-electroencephalography data. The neuron counts of the lateral amygdaloid nucleus did not correlate with various presumed etiological factors of TLE including hereditary seizures, birth complications, febrile convulsions, traumatic brain injury, infections, seizure semiology, and epileptological outcome. However, patient age at surgery was significantly higher (mean difference 10 years) when AS was present, as compared with patients without AS (p < 0.01). Seizure origin, as determined by using amygdalohippocampal depth electrodes, did not correlate with the presence or absence of AS. Neuropsychologically, there was a significant correlation between the neuronal densities of the lateral amygdaloid nucleus and both preoperative visual recognition and postoperative deterioration of short-term verbal memory performance (p < 0.05). CONCLUSIONS: Except for the relatively long history of epilepsy, the presence of AS is not associated with specific clinical or electrocorticographic features of mesial TLE. However, patients without AS are particularly at risk for deterioration of short-term verbal memory following amygdalohippocampectomy. PMID- 10389882 TI - Thalamic deep brain stimulation for the treatment of head, voice, and bilateral limb tremor. AB - OBJECT: In published series of patients who undergo deep brain stimulation (DBS) of the thalamus the effects of unilateral stimulation on contralateral limb tremor have been reported. The authors detail their experience with bilateral thalamic DBS in the treatment of head, voice, and bilateral limb tremor and compare it with earlier studies of unilateral stimulation. METHODS: Twenty-three patients (six with Parkinson's disease, 15 with essential tremor, and two with multiple sclerosis) underwent 19 bilateral DBS procedures (nine staged, 10 simultaneous) and four procedures contralateral to thalamotomy to control tremor of the head in 10, voice in seven, and limbs in 20 patients. Limb tremor improvement was graded as follows: 4, no tremor; 3, stress-induced tremor; 2, functional improvement; 1, no functional improvement; and 0, persistent tremor. Improvement of head or voice tremor was graded as follows: 4, greater than 75%; 3, between 50% and 75%; 2, between 25% and 50%; 1, less than 25%; and 0, no improvement. The mean follow-up period was 10 months. Twenty-two patients (96%) demonstrated improved tremor at the last follow-up review. Of 20 patients with bilateral limb tremor, 17 (85%) improved to Grades 3 and 4, two patients (10%) with multiple sclerosis improved to Grade 2, and one (5%) exhibited tremor recurrence 8 months later. Nine (90%) of 10 patients with severe head tremor improved to Grades 4 or 3. Six (86%) of seven patients with voice tremor improved to Grade 3. Seven patients (30%) developed dysarthria, and seven (30%) developed disequilibrium; symptoms reversed in the majority of patients after the stimulation parameters were changed. One patient (4%) developed mild memory decline. There were no deaths. CONCLUSIONS: The following findings are reported: 1) bilateral thalamic DBS and stimulation contralateral to thalamotomy are safe; 2) staging the procedure does not reduce the risk of dysarthria or gait disequilibrium; and 3) head and voice tremor are primary indications for bilateral DBS. PMID- 10389883 TI - Functional neuronavigation with magnetoencephalography: outcome in 50 patients with lesions around the motor cortex. AB - OBJECT: The authors conducted a study to evaluate the clinical outcome in 50 patients with lesions around the motor cortex who underwent surgery in which functional neuronavigation was performed. METHODS: The sensorimotor cortex was identified in all patients with the use of magnetoencephalography (MEG). The MEG source localizations were superimposed onto a three-dimensional magnetic resonance image and the image data set was implemented into a neuronavigation system. Based on this setup, the surgeon chose the best surgical strategy. During surgery, the pre- and postcentral gyri were identified by neuronavigation and, in addition, the central sulcus was localized using intraoperative recording of somatosensory evoked potentials. In all cases MEG localizations of the sensory or motor cortex were correct. In 30% of the patients preoperative paresis improved, in 66% no additional deficits occurred, and in only 4% (two patients) deterioration of neurological function occurred. In one of these patients the deterioration was not related to the procedure. CONCLUSIONS: The method of incorporating functional data into neuronavigation systems is a promising tool that can be used in more radical surgery to lessen morbidity around eloquent brain areas. PMID- 10389884 TI - Human arachnoid villi response to subarachnoid hemorrhage: possible relationship to chronic hydrocephalus. AB - OBJECT: The origin of chronic communicating hydrocephalus following subarachnoid hemorrhage (SAH) is not well understood. Fibrosis of the arachnoid villi has been suggested as the cause for obstruction of cerebrospinal fluid (CSF) flow, but this is not well supported in the literature. The goal of this study was to determine the relationship between blood, inflammation, and cellular proliferation in arachnoid villi after SAH. METHODS: Arachnoid villi from 50 adult patients were sampled at autopsy. All specimens were subjected to a variety of histochemical and immunohistochemical stains. The 23 cases of SAH consisted of patients in whom an autopsy was performed 12 hours to 34 years post-SAH. Fifteen cases were identified as moderate-to-severe SAH, with varying degrees of hydrocephalus. In comparison with 27 age-matched non-SAH controls, the authors observed blood and inflammation within the arachnoid villi during the 1st week after SAH. Greater mitotic activity was also noted among arachnoid cap cells. The patient with chronic SAH presented with ventriculomegaly 2 months post-SAH and exhibited remarkable arachnoid cap cell accumulation. CONCLUSIONS: The authors postulate that proliferation of arachnoidal cells, triggered by the inflammatory reaction or blood clotting products, could result in obstruction of CSF flow through arachnoid villi into the venous sinuses. This does not exclude the possibility that SAH causes generalized fibrosis in the subarachnoid space. PMID- 10389885 TI - Inhibition of NF2-negative and NF2-positive primary human meningioma cell proliferation by overexpression of merlin due to vector-mediated gene transfer. AB - OBJECT: The absence of in vitro models of neurofibromatosis Type 2 (NF2) defective meningiomas has limited investigative efforts to study the biological effects of this gene in the pathogenesis of these tumors. The goals of this report are to show that gene transfer vectors can efficiently express the wild type NF2 transgene into primary meningioma cells and to determine effects on cellular proliferation. METHODS: In this study, the authors have compared the transducing capacities of a retrovirus, an adenovirus, and a herpes simplex virus amplicon vector for use in primary human meningioma cells harvested from human tumors excised from patients with and without NF2. Transduction efficiencies with the latter vector approached 100% and it was selected to transfer the wild-type NF2 transgene into these cells. Western blot analysis confirmed that vector mediated gene transfer mediated the expression of the NF2-encoded polypeptide merlin. Overexpression of merlin significantly inhibited the proliferation of both NF2-negative and NF2-positive human meningioma cells when compared to the proliferation of cells transduced with a control vector. CONCLUSIONS: This study demonstrates the feasibility of using vector-mediated gene transfer to study wild type NF2 gene function in short-term cultures of primary human meningioma cells. PMID- 10389886 TI - Growth hormone receptor expression and function in meningiomas: effect of a specific receptor antagonist. AB - OBJECT: This study was undertaken to explore the effects of growth hormone (GH) and the GH-stimulated peptide insulin-like growth factor-1 (IGF-1) on the growth rate of meningiomas. METHODS: Polymerase chain reaction and ribonuclease protection assays were used to demonstrate that GH receptor messenger RNA was present in all 14 meningioma specimens studied, regardless of tumor grade. Both wild type (GHRwt) and a previously described exon 3 deletion isoform (GHRd3) of the GH receptor were identified in individual tumor specimens. The importance of the GH receptor was assessed using a GH receptor antagonist (B2036). Blockade of the GH receptor with B2036 reduced serum-induced DNA synthesis, as measured by thymidine incorporation, by 8 to 33% (mean 20%) in primary meningioma cultures. Tumors that expressed the GHRwt and GHRd3 isoforms, or a combination of the two, were all responsive to antagonist treatment. The importance of IGF- in stimulating meningioma cell growth was also assessed. It was found that IGF-1 increased thymidine incorporation in primary meningioma cultures in a dose dependent manner: 1 ng/ml, 5 ng/ml, and 10 ng/ml resulted in increases in thymidine incorporation of 21%, 43%, and 176%, respectively, over baseline values. CONCLUSIONS: In these studies the authors demonstrate that activation of the GH/IGF-1 axis significantly increases the growth rate of meningiomas. Blockade of the GH receptor on tumor cells inhibited tumor growth. If these findings are confirmed in animal studies, agents that downregulate the GH/IGF-1 axis might represent a potential adjuvant therapy in the management of patients with meningioma. PMID- 10389887 TI - Long-term effects of in vivo angioplasty in normal and vasospastic canine carotid arteries: pharmacological and morphological analyses. AB - OBJECT: A canine model of hemorrhagic vasospasm of the high cervical internal carotid artery (ICA) was used to study the long-term effects of transluminal balloon angioplasty (TBA) on the structure and function of the arterial wall. METHODS: Forty dogs underwent surgical exposure of both distal cervical ICAs, followed by baseline angiographic studies on Day 0. Dogs in Group A (20 animals) underwent simple exposure of one ICA and placement of a silicone elastomer cuff around a segment of the opposite artery. These animals underwent repeated angiography on Day 7, and then TBA was performed on the uncuffed ICA; the cuff was removed from the opposite vessel. For dogs in Group B (20 animals), blood clot-filled cuffs were placed around both ICAs, and on Day 7 angiography was repeated and TBA was performed on one randomly selected ICA. Four animals were then killed from each group, and in the remaining animals the cuffs were removed from both ICAs. On Days 14, 21, 28, and 56, four animals from each group underwent repeated angiography and were then killed to permit pharmacological and morphological analyses of the ICAs. This protocol yielded five study categories: cuffed nonblood-coated arteries not subjected to TBA, blood-coated arteries not subjected to TBA, blood-coated arteries subjected to TBA, normal arteries subjected to TBA, and control arteries obtained from the proximal ICA in each animal. The contractile responses of isolated arterial rings obtained from each ICA were recorded after treatment with potassium chloride, noradrenaline, and serotonin, whereas relaxations in response to the calcium ionophore A23187 and papaverine were recorded after tonic contraction to noradrenaline had been established. Morphological analysis was performed using scanning electron microscopy. Arteries surrounded by an empty cuff exhibited no angiographic, pharmacological, or morphological differences compared with normal arteries on any study day. Arteries surrounded by blood developed angiographically confirmed vasospasm on Day 7, with characteristic pharmacological and morphological features; resolution of these symptoms occurred by Day 21. Vasospastic arteries subjected to TBA on Day 7 remained dilated on angiographic studies, exhibited impaired responses to pharmacological agents (except for papaverine), and showed altered morphological features until Day 28. Normal arteries subjected to TBA on Day 7 remained dilated on angiographic studies, exhibited impaired responses to pharmacological agents (except for papaverine), and displayed altered morphological features until Day 14. CONCLUSIONS: These results indicate that the canine high cervical ICA model produces consistent and reproducible vasospasm that follows a similar time course to that seen in humans. When TBA is performed in vasospastic arteries, it results in an immediate functional impairment of vascular smooth muscle that lasts for 2 weeks, with resolution at 3 weeks; morphological changes are mostly resolved 3 weeks post-TBA. In normal vessels, TBA causes functional impairment and morphological alterations that are not as severe or as long-lasting as those seen in vasospastic arteries. PMID- 10389888 TI - Immunosuppression for neural xenografts: a comparison of cyclosporin and anti CD25 monoclonal antibody. AB - OBJECT: The goal of this study was to compare the effects of short- and long-term immunosuppression induced by cyclosporin with those of immunosuppression induced by a monoclonal antibody against the rat interleukin-2 receptor (anti-CD25 mAb) in rats with xenografts. METHODS: The authors compared the in vivo function and final histological characteristics of fetal mouse mesencephalon xenografts in hemiparkinsonian rats in which immunosuppression was induced by: 1) a short course (2 weeks) of cyclosporin; 2) a long course (8 weeks) of cyclosporin; or 3) a short course of treatment with anti-CD25 mAb. Adult Wistar rats were unilaterally lesioned with 6-hydroxydopamine in their medial forebrain bundle, after which their rotational behavior in response to methamphetamine was quantified. Four groups of 20 rats with rotations numbering greater than six turns per minute received fetal mouse mesencephalon transplants to their dopamine denervated striatum. Group 1 received no immunosuppression therapy; Group 2 received daily intraperitoneal injections of 10 mg/kg cyclosporin for 2 weeks; Group 3 received daily intraperitoneal injections of 10 mg/kg cyclosporin for 8 weeks; and Group 4 received daily intraperitoneal injections of 1 mg/kg anti-CD25 mAb for 2 weeks. The rats were tested for rotational behavior every 4 weeks and killed after 16 weeks. Surviving xenografts were assessed using immunohistochemical staining for a mouse neuronal marker (Thy-1.2). Sixteen weeks after transplant, there were significantly more surviving xenografts in Groups 3 (p < 0.001) and 4 (p < 0.001) compared with control Group 1 (Fisher's exact test) and significantly better functioning xenografts in Groups 3 (p < 0.01) and 4 (p < 0.05) compared with control Group 1 (contrasts of groups following analysis of variance with Bonferroni correction). CONCLUSIONS: A short course of anti-CD25 mAb-induced immunosuppression was as effective as a long course of cyclosporin induced immunosuppression in this model. PMID- 10389889 TI - Influence of rewarming conditions after hypothermia in gerbils with transient forebrain ischemia. AB - OBJECT: Recently, several studies have demonstrated that hypothermia has a beneficial effect on clinical outcome; however, it is difficult to determine the appropriate rewarming conditions in clinical use. The purpose of the present study was to examine the influence of rewarming conditions in gerbils with transient forebrain ischemia. METHODS: Ischemia was induced in the gerbils by a 5 minute bilateral common carotid artery occlusion, after which the animals were immediately subjected to moderate or deep hypothermia. After moderate hypothermia (30.5 degrees C for 4 hours) the animals were rewarmed over standard, fast, or slow time periods. After deep hypothermia (24 degrees C for 2 hours) the animals were rewarmed in a standard, fast, slow, or stepwise manner. Cerebral blood flow (CBF), extracellular glutamate, and lactate were monitored. Hippocampal CA I cell damage was assessed 7 days after induction of ischemia. In animals treated with moderate hypothermia, the rewarming rate had no influence on the number of surviving neurons. However, fast rewarming from deep hypothermia (to 37 degrees C for 30 minutes) failed to provide the neuroprotective effect of hypothermia. Furthermore, this group showed a poor recovery of CBF (p < 0.01) and, consequently, an increase in extracellular glutamate (p < 0.01) and lactate (p < 0.01) in the hippocampus. CONCLUSIONS: The results of this study indicate a transient uncoupling of CBF and cerebral metabolism during fast rewarming from deep hypothermia, whereas slow and stepwise rewarming periods were found to be useful for protection against uncoupling of CBF and cerebral metabolism during rewarming. PMID- 10389890 TI - Painful supernumerary phantom arm following motor cortex stimulation for central poststroke pain. Case report. AB - In this report, the authors describe a case in which the patient began to experience a supernumerary phantom arm after she received motor cortex stimulation for central pain. The patient had a history of right thalamocapsular stroke. It is speculated that the motor cortex activation triggered a response in the patient's parietal lobe, precipitating perception of the phantom limb. To the authors' knowledge this is the first reported case of its kind. PMID- 10389891 TI - Postoperative swelling of pericranial pedicle graft producing intracranial mass effect. Report of two cases. AB - Two cases of florid swelling of pericranial pedicle grafts are reported. Intracranial mass effect produced by the grafts necessitated reoperation with graft removal in one case and graft revision in the other. No permanent neurological deficits were incurred by either patient. Venous congestion and associated swelling within the graft were considered to be related to constriction of the graft base at the frontal bone flap-skull base interface in one patient and torsion of the graft base in the other. PMID- 10389892 TI - Rare occurrence of intracerebellar colloid cyst. Case report. AB - Colloid cysts are rare intracerebral lesions that are preferentially encountered within the third ventricle. There are only a few reports in which colloid cysts are described in other locations such as the fourth ventricle. A symptomatic intracerebellar colloid cyst in a 45-year-old woman is described. The patient presented with headache, gait disturbance, and nausea. Neuroradiological imaging revealed compression of the fourth ventricle, hydrocephalus, and an intracerebellar cystic lesion measuring 4 x 5 cm that had a small peripheral solid portion. The cyst was successfully removed via a paramedian suboccipital approach. Postoperatively, the patient recovered quickly. The findings in the present report represent an additional example of the broad spectrum of cystic lesions encountered in the cerebellum. PMID- 10389893 TI - Falcotentorial plasmacytoma. Case report. AB - Intracranial solitary plasmacytomas are extremely rare tumors and are often misdiagnosed preoperatively. The authors report the successful treatment of a patient who harbored such a tumor involving both the falx and tentorium; this is the second case reported. A 59-year-old woman suffered from a seizure disorder due to a falcotentorial lesion, which had been identified 3 years earlier and was thought at the time to be an en plaque meningioma. Most recently, the patient presented with symptoms of increased intracranial pressure and hemiparesis. Computerized tomography and magnetic resonance imaging of her head revealed progressive growth of the tumor. The patient underwent partial resection of the tumor and chemo- and radiation therapies. Intracranial plasmacytomas must always be included in a differential diagnosis because potential complete cure can be achieved using fairly conservative treatment modalities. PMID- 10389894 TI - An intracranial aneurysm on the feeding artery of a cerebellar hemangioblastoma. Case report. AB - A case of cerebellar hemangioblastoma with a coexistent arterial aneurysm on the feeding artery of the tumor is reported. The patient presented with an acute onset of headache, loss of consciousness, and left-sided hemiparesis due to a posterior fossa hemorrhage found adjacent to a hemangioblastoma. Four-vessel angiography revealed an aneurysm on the anterior inferior cerebellar artery (AICA), which was the main feeding vessel of the hemangioblastoma. Successful total excision of the hemangioblastoma and clipping of the AICA aneurysm achieved in a one-stage operation was demonstrated on postoperative angiography. PMID- 10389895 TI - Fusiform vertebral artery aneurysms as a cause of dissecting aneurysms. Report of two autopsy cases and a review of the literature. AB - Two autopsy cases of angiographically determined fusiform aneurysms of the vertebral arteries (VAs) are reported and the appropriate literature is reviewed to investigate the pathological characteristics of both fusiform and dissecting VA aneurysms and the pathogenesis of dissecting aneurysms. One patient had suffered a subarachnoid hemorrhage (SAH) due to dissection of a previously documented incidental fusiform aneurysm. The other patient had harbored incidental fusiform aneurysms coexistent with a ruptured aneurysm of the posterior inferior cerebellar artery. The location and pathological features of the aneurysms were similar in the two cases. The aneurysms in both cases displayed intimal thickening, disruption of the internal elastic lamina, and degeneration of the media. A mural hemorrhage and patchy calcification were also found in the case that included SAH. Based on their pathological investigation of these two cases and a review of reported cases, the authors propose that incidental fusiform aneurysms in the VAs are characterized by weakness in the internal elastic lamina and, therefore, have the potential to become dissecting aneurysms, resulting in a fatal prognosis. This suggests that long-term control of blood pressure is mandatory in patients with incidental fusiform aneurysms in the VAs. PMID- 10389896 TI - Computer-generated microsurgical anatomy of the basilar artery bifurcation. Technical note. AB - The authors' goal was to develop a computer graphics model to represent the microsurgical anatomy of the basilar artery (BA) bifurcation and surrounding structures to simulate surgery of a BA bifurcation aneurysm performed via the transsylvian approach. The source of the input data was a variety of publications that showed detailed anatomy of the area. A computer graphics model of the area near the BA bifurcation including relevant structures, such as perforating branches or cranial nerves, was depicted in detail. A BA bifurcation aneurysm was added to the computer graphics model and it was rotated to simulate the transsylvian approach. After the internal carotid artery was displaced using a virtual retractor, the aneurysm was exposed, thus providing an understanding of the three-dimensional surgical orientation of the area. Designing a standard anatomical model on the basis of data culled from a variety of publications and adding morphological changes by using a virtual retractor to displace structures that obstruct the view along a critical path at the base of the brain are useful strategies of computer manipulation for surgical simulation in open microneurosurgery. This methodological tool would be useful in teaching surgical microanatomy and in introducing a new navigational system for virtual reality. Both concept and technical details are discussed. PMID- 10389897 TI - Balloon angioplasty of the A1 segment of the anterior cerebral artery narrowed by vasospasm. Technical note. AB - The authors describe a new endovascular technique that improves catheterization and balloon angioplasty of the A1 segment of the anterior cerebral artery after it has been narrowed by vasospasm. The technical results of using this method in seven patients are presented. PMID- 10389898 TI - Intraoperative electrooculographic monitoring of oculomotor nerve function during skull base surgery. Technical note. AB - Intraoperative monitoring techniques for protecting the integrity of the oculomotor nerves during skull base surgery have been reported by several investigators, all of which involved the use of electromyographic responses to extraocular muscles. However, these techniques have not yet become popular because of the complexity of the procedures. The authors report an extremely simple and far more reliable technique in which electrooculographic (EOG) monitoring is used. The oculomotor nerves were stimulated with a monopolar electrode during skull base exposure. The polarity of the EOG responses recorded with surface electrodes placed on the skin around the eyeball yielded precise information concerning the location and function of the oculomotor and abducent nerves. In addition, with the aid of continuous EOG monitoring that detected transient changes in the background waves, surgical procedures that might impinge on oculomotor nerve function could be avoided. The present technique has been used in eight patients with skull base tumors and with it, the authors have achieved excellent results. PMID- 10389899 TI - Giant aneurysm associated with a large cyst. Case illustration. PMID- 10389900 TI - Hypoglossal-facial nerve side-to-end anastomosis. PMID- 10389901 TI - Blisterlike aneurysms. PMID- 10389902 TI - Functional positron emission tomography. PMID- 10389903 TI - Intraventricular hemangiopericytoma. PMID- 10389904 TI - High jugular bulb oxygen saturation. PMID- 10389905 TI - Sylvian aqueduct syndrome. PMID- 10389906 TI - Expression of transforming growth factor beta type II receptors in head and neck squamous cell carcinoma. AB - Transforming growth factor (TGF)-beta is a potent regulator of growth and differentiation in normal squamous epithelium. TGF-beta exerts its antiproliferative effect via the TGF-beta type II receptor (TbetaR-II). A decrease in TbetaR-II expression is believed to be responsible, in part, for the resistance of squamous cell carcinoma (SqCC) to the anti-proliferative effects of TGF-beta. In the present study, we used immunohistochemistry and in situ hybridization to analyze the expression of TbetaR-II along the successive oncogenic stages of head and neck squamous neoplasia, from normal epithelium to dysplasia to carcinoma. Quantitation of TbetaR-II expression in 38 SqCCs was assessed on a visual scale ranging from negative (absence of staining) to 3+ (strong staining). Normal squamous epithelium and squamous epithelium in the vicinity of the tumors showed homogenous receptor expression with moderate intensity. Dysplastic epithelium and carcinoma in situ showed a mild decrease in receptor expression intensity. Well-differentiated to moderately differentiated carcinomas showed heterogeneous expression of variable intensity, and poorly differentiated carcinomas were completely devoid of TbetaR-II. In every tumor, the superficial component showed more intense receptor expression than the invasive component. These results indicate that TbetaR-II expression inversely correlates with disease aggressiveness and suggest that aberrant TbetaR-II expression is a contributing factor to the pathogenesis of SqCC. PMID- 10389907 TI - Reduction of BRCA1 protein expression in Japanese sporadic breast carcinomas and its frequent loss in BRCA1-associated cases. AB - BRCA1 is a tumor suppressor gene that is responsible for hereditary breast and ovarian cancer syndrome. To clarify the possible involvement of the BRCA1 protein in mammary carcinogenesis in sporadic and hereditary forms, we have analyzed the BRCA1 protein expression pattern in five breast epithelial cell lines, including a BRCA1-deficient cell line, and 162 breast cancer tissue samples [including 108 sporadic, 35 hereditary (BRCA1 status unknown), and 19 BRCA1-associated cases] from Japanese women. Twelve anti-BRCA1 antibodies were tested by fixation conditions, in which nuclear localization of BRCA1 protein was preserved, and by specificity of the antibodies, which was evaluated in BRCA1-deficient cancer cells. Using monoclonal antibodies applicable to immunohistochemical analysis of paraffin-embedded tissue sections, we found high-level expression of BRCA1 protein in normal mammary epithelium and various degrees of reduced expression in breast cancer cells. Of the 19 BRCA1-associated breast cancer tissues, 15 (79%) showed reduction (8 cases) or complete loss (7 cases) of nuclear expression. Thirty (28%) of 108 sporadic and 6 (17%) of 35 hereditary carcinomas showed reduced BRCA1 protein expression. Reduction of BRCA1 protein expression in sporadic carcinomas was associated with solid-tubular phenotype, with poor tubular differentiation, and with an overexpression of c-erbB-2 protein, which is one of the prognostic factors in breast cancer. Our data suggest that reduced expression of BRCA1 protein may play an important role in mammary carcinogenesis, not only in BRCA1-associated breast carcinomas, but also in sporadic carcinomas, and also suggest that mechanisms other than mutation may be involved in its reduced expression. PMID- 10389908 TI - Spontaneous ex vivo apoptosis of peripheral blood mononuclear cells in patients with head and neck cancer. AB - Proportions of apoptotic (TUNEL+) peripheral blood mononuclear cells (PBMCs) were measured by flow cytometry in patients with head and neck cancer and normal controls at the time of blood draws (0 time) and after 24-h incubation. PBMCs were incubated at 37 degrees C in medium (spontaneous apoptosis) and in the presence of CH-11 antibody (anti-Fas) or tumor necrosis factor (TNF)-alpha, both capable of inducing DNA fragmentation in activated T cells expressing the TNF family of receptors. PBMCs obtained from the patients had significantly higher (P < 0.0001) proportion of apoptotic cells than PBMCs of controls at 0 time as well as after 24-h incubation. Ex vivo apoptosis included all subsets of PBMCs: CD3+ T cells, CD16+ CD56+ natural killer cells, CD19+ B cells, and CD14+ monocytes, as determined by two-color flow cytometry. However, T cells represented the largest PBMC subset undergoing apoptosis, and lymphocytes rather than monocytes were the major TUNEL+ PBMC population. Among T cells, the level of spontaneous ex vivo apoptosis was nearly as high as that of CH-11 antibody-induced or TNF-alpha induced apoptosis, indicating that activated Fas+ and TNFR1+ T cells were preprogrammed in vivo to die. Also, elevated levels of spontaneous apoptosis at time 0 in patients with head and neck cancer (P < 0.0001) indicated that a higher fraction of PBMCs was undergoing apoptosis in vivo in patients than controls. Together, the data suggest that an increased rate of turnover of lymphocytes is associated with cancer and may be responsible for functional lymphocyte imbalance, even in treated patients who have no evident disease. PMID- 10389909 TI - Detection of subclinical cancers by prostate-specific antigen screening in asymptomatic men from high-risk prostate cancer families. AB - Positive family history is a significant risk factor for prostate cancer. Improved knowledge of the epidemiology and molecular basis of hereditary prostate cancer has led to a need for counseling and clinical follow-up for men with a positive family history of prostate cancer. However, very little information is available on the efficacy of early screening procedures, such as serum prostate specific antigen (PSA) measurements, in the management of genetically predisposed, high-risk individuals. In a nationwide study, we obtained family histories from 2099 Finnish prostate cancer patients and identified 302 families with two or more affected cases. Here, 209 asymptomatic 45-75-year-old males from these families were included in a study to determine the frequency of serum PSA positivity and the prevalence of subclinical prostate cancers. Serum PSA was elevated in 21 (10.0%) of these high-risk individuals. Seven prostate cancers (3.3%) and two high-grade prostatic intraepithelial neoplasia lesions were diagnosed, with three cancers occurring in men ages < or = 59 years. Men from prostate cancer families with an average age of onset of < 60 years had a significantly higher frequency of PSA positivity (28.6%, P = 0.01) as well as cancers (14.3%, P = 0.02) than those with a later age of onset. The results suggest that prostate cancer development in genetically predisposed individuals is preceded by a subclinical period when PSA detection is possible. Serum PSA screening may be particularly useful in men with a family history of early-onset prostate cancer. PMID- 10389910 TI - Generation of anti-p53 cytotoxic T lymphocytes from human peripheral blood using autologous dendritic cells. AB - CTLs recognizing the HLA-A2.1-restricted, wild-type sequence p53 epitopes p53(149 157) and p53(264-272) were generated from CD8-enriched populations of nonadherent peripheral blood lymphocytes (PBLs) obtained from healthy donors. The PBLs were restimulated in vitro with peptide-pulsed granulocyte macrophage colony stimulating factor- and interleukin (IL)-4-induced autologous dendritic cells in the presence of IL-6 and IL-12 and subsequently cultivated with IL-1alpha, IL-2, IL-4, IL-6, and IL-7. Bulk anti-p53(264-272) CTL populations were generated from PBLs obtained from two of five donors. Both CTL populations were cytotoxic against peptide-pulsed HLA-A2+ target cells, but not against untreated target cells. A CD8+ anti-p53 CTL clone designated p264#2 was isolated from one of the bulk populations. It was found to have an intermediate affinity of approximately 10(-9) M for the epitope and to mediate cytotoxicity against several human tumor cell lines, including the squamous cell carcinoma of the head and neck cell line SCC-9, which is known to present the wild-type sequence p53(264-272) epitope. In addition, CTLs reactive against p53(149-157)-pulsed targets as well as a HLA-A2+ tumor cell line were cloned from a bulk population of antitumor CTLs obtained from one of the five normal PBLs restimulated with this epitope. The results indicate that CTLs recognizing wild-type sequence epitopes can be generated from precursors present in PBLs obtained from some normal individuals using autologous dendritic cells as antigen-presenting cells and suggest that vaccine strategies targeting these epitopes can lead to antitumor CTL generation, thereby emphasizing the therapeutic potential of p53-based cancer vaccines. PMID- 10389911 TI - Generation of immunity to the HER-2/neu oncogenic protein in patients with breast and ovarian cancer using a peptide-based vaccine. AB - HER-2/neu is a "self" tumor antigen that is overexpressed in 15-30% of human adenocarcinomas. Vaccine strategies directed against HER-2/neu and other self tumor antigens require development of methods to overcome immune tolerance to self-proteins. In rats, rat neu peptide vaccines have been shown to be an effective way of circumventing tolerance to rat neu protein and generating rat neu-specific immunity. The present report validates that a similar peptide-based vaccine formulation is effective for inducing T-cell immunity to HER-2/neu protein in humans with breast and ovarian cancer. The vaccine formulation included groups of peptides derived from the HER-2/neu extracellular domain (ECD) or intracellular domain (ICD) mixed with granulocyte macrophage colony stimulating factor as an adjuvant. These peptides were 15-18 amino acids in length and designed to elicit a CD4 T helper-specific immune response. Patients underwent intradermal immunization once a month for a total of two to six immunizations. To date, all of the patients immunized with HER-2/neu peptides developed HER-2/neu peptide-specific T-cell responses. The majority of patients (six of eight) also developed HER-2/neu protein-specific responses. Responses to HER-2/neu protein occurred with epitope spreading. Immune T cells elicited by vaccination were shown to migrate outside the peripheral circulation by virtue of generating delayed type hypersensitivity responses distant from the vaccine site, which indicated the potential ability to traffic to the site of tumor. The use of peptide-based vaccines may be a simple, yet effective, vaccine strategy for immunizing humans to oncogenic self-proteins. PMID- 10389912 TI - A Phase I study of continuous infusion doxorubicin and paclitaxel chemotherapy with granulocyte colony-stimulating factor for relapsed epithelial ovarian cancer. AB - A Phase I study of paclitaxel and doxorubicin administered as concurrent 96-h continuous i.v. infusion was performed to determine the maximum tolerated dose (MTD), principal toxicities, and pharmacokinetics of this combination in women with relapsed epithelial ovarian cancer. The paclitaxel dose was fixed at 100 mg/m2 (25 mg/m2/day for 4 days). The dose of doxorubicin was escalated from 30 mg/m2 (7.5 mg/m2/day for 4 days) in increments of 10 mg/m2 until dose-limiting toxicity was observed. All patients received granulocyte colony-stimulating factor 5 microg/kg/day prophylactically. Apparent steady-state plasma levels of both drugs were determined in the final cohort of patients treated at the MTD. A total of 17 patients received 52 cycles of therapy. The median age was 58 years, and all patients had previously received one to five different regimens (median, 2) of chemotherapy, including both platinum and paclitaxel. The treatment was tolerated well, with grade 1-2 nausea being the most frequent side effect (73% of cycles). Anemia, neutropenia, thrombocytopenia, and mucositis became dose limiting at the fourth dose level, defining the MTD of doxorubicin in this regimen as 50 mg/m2. There were four partial responses and one complete response in 15 evaluable patients. Apparent steady-state plasma concentrations (mean +/- SD) of paclitaxel and doxorubicin in the three patients treated at the MTD were 33.9 +/- 12.5 nM and 15.7 +/- 1.3 nM, respectively. Paclitaxel and doxorubicin by continuous infusion is a well-tolerated and active chemotherapy regimen for recurrent ovarian cancer. PMID- 10389913 TI - Rhenium-186-labeled hydroxyethylidene diphosphonate dosimetry and dosing guidelines for the palliation of skeletal metastases from androgen-independent prostate cancer. AB - Rhenium-186 (tin)-labeled hydroxyethylidene diphosphonate (186Re-labeled HEDP) was evaluated in 27 men with progressive androgen-independent prostate cancer and bone metastases. Administered activities ranged from 1251 to 4336 MBq (33.8-117.2 mCi). The primary objectives were to assess tumor targeting, normal organ dosimetry, and safety. Antitumor effects were assessed by posttherapy changes in prostate-specific antigen and, when present, palliation of pain. Whole-body kinetics, blood and kidney clearance, skeletal dose, marrow dose, and urinary excretion of the isotope were assessed. Targeting of skeletal disease was observed over the period of quantification (4-168 h). Radiation doses to whole body, bladder, and kidney were well tolerated. The dose-limiting toxicity was myelosuppression (grade III) at 4107 MBq (111 mCi) and grade II at 296 MBq (80 mCi). Probe clearance (whole body) and urinary excretion measurements were highly correlated. Of the six patients treated at the highest dosage schedules (three at 1510 MBq/m2 and three at 1665 MBq/m2), three showed a posttherapy decline in prostate-specific antigen of 50% or more. The declines were not sustained. The determination of total activity retained at 24 h, as well as an estimate of marrow dose, correlated with the amount of myelosuppression observed. These results suggest that a single 24-h measurement of retained activity would allow individualized dosing and an improved therapeutic index relative to fixed dosing schema. Repetitive dosing is required to increase palliation. PMID- 10389914 TI - Long survival of patients with small cell lung cancer after adjuvant treatment with the anti-idiotypic antibody BEC2 plus Bacillus Calmette-Guerin. AB - Despite active therapies for small cell lung cancer (SCLC), most patients relapse and die of the disease. The present study evaluates immunization using the anti idiotypic antibody BEC2, which mimics the ganglioside GD3 expressed on the surface of most SCLC tumors, combined with Bacillus Calmette-Guerin (BCG) as an immune adjuvant. We hypothesized that active immunization could alter the natural history of the disease. Fifteen patients who had completed standard therapy for SCLC received a series of five intradermal immunizations consisting of 2.5 mg of BEC2 plus BCG over a 10-week period. Blood was collected for serological analysis, and outcome was monitored. All patients developed anti-BEC2 antibodies, despite having received chemotherapy with or without thoracic radiation. We detected anti-GD3 antibodies in five patients, including those with the longest relapse-free survival. The median relapse-free survival for patients with extensive stage disease is 11 months and has not been reached for patients with limited stage disease (>47 months), with only one of seven patients having relapsed after a median follow-up of 47 months. Immunization of patients with SCLC after standard therapy using BEC2 plus BCG can induce anti-GD3 antibodies and is safe. The survival and relapse-free survival in this group of patients are substantially better than those observed in a prior group of similar patients. A Phase III trial is being conducted to evaluate BEC2 plus BCG as adjuvant therapy after chemotherapy and irradiation. PMID- 10389915 TI - Pharmacokinetics and pharmacodynamics of 9-aminocamptothecin infused over 72 hours in phase II studies. AB - A novel derivative of camptothecin, 9-aminocamptothecin (9-AC), is currently under Phase II evaluation in various cancers. Exceptionally mild toxicities were observed in patients with brain tumors who were treated with anticonvulsants. To investigate a pharmacokinetic interaction between 9-AC and anticonvulsants, and to evaluate the pharmacodynamics of 9-AC, we investigated the clinical pharmacology of 9-AC, administered by a 72-h infusion, in three Phase II studies. Plasma concentrations of total 9-AC (lactone plus carboxylate) at a steady state were measured in 56, 10, and 14 patients with non-small cell lung cancer, malignant glioma, and head and neck cancer, respectively. For lung cancer or glioma patients, 9-AC was infused at 45 (51 patients) or 59 (15 patients) microg/m2/h, and 9-AC was infused at 35.4 microg/m2/h in head and neck cancer patients. All glioma patients had been treated with phenytoin or carbamazepine. 9 AC clearance did not differ among the dosage rates, but differed according to the diseases (P = 0.002). Glioma patients had a higher clearance (1.0-18.0; median, 2.0 liters/h/m2) than lung cancer patients (0.3-5.1; median, 0.9 liters/h/m2). A logistic regression model described the relationship between the 9-AC concentration and the probability of grade 4 neutropenia, which was the main toxicity. Observed incidences of grade 4 neutropenia for patients with model predicted probability of 0-20%, 20-40%, and 40-100% were 10%, 32%, and 67%, respectively, and corresponded to 9-AC concentration of <54, 54-86, and >86 ng/ml, respectively. Anticonvulsants seem to induce the clearance of 9-AC, and the concentration of 9-AC predicts the probability of grade 4 neutropenia. PMID- 10389916 TI - A Phase I study of active immunotherapy with carcinoembryonic antigen peptide (CAP-1)-pulsed, autologous human cultured dendritic cells in patients with metastatic malignancies expressing carcinoembryonic antigen. AB - Dendritic cells (DCs), antigen-presenting cells capable of priming naive T cells to specific antigens in an HLA-restricted fashion, have been demonstrated to induce protective T cell-mediated immunity in tumor-bearing animals. We performed this study to test the safety, feasibility, and clinical response of immunizations with in vitro-generated DCs, loaded with an HLA-A2-restricted peptide fragment of the tumor antigen carcinoembryonic antigen (CEA), for the treatment of patients with advanced CEA-expressing malignancies. Cell preparations enriched for autologous DCs were generated from the patients' plastic adherent peripheral blood mononuclear cells in serum-free media supplemented with granulocyte macrophage colony-stimulating factor and interleukin-4. Within the cell preparation, 66% of the cells expressed the phenotype typical for DCs (CD86high, HLA-DRhigh, and CD14low). The DCs were loaded with the CEA peptide CAP-1 and cryopreserved. Groups of three to six patients received four weekly or biweekly i.v. infusions of the CAP-1-loaded DC in escalating dose levels of 1 x 10(7), 3 x 10(7), and 1 x 10(8) cells/dose. A subset of the patients in the last group also received intradermal injections of 1 x 10(6) DCs. There were no toxicities directly referable to the treatments. One patient had a minor response, and one had stable disease. Skin punch biopsy at DC injection sites demonstrated pleomorphic infiltrates in the three patients evaluated. We conclude that it is feasible and safe to generate and administer large numbers of previously cryopreserved DCs loaded with CAP-1 peptide to patients with advanced malignancies. PMID- 10389917 TI - A Phase I trial of calcitriol (1,25-dihydroxycholecalciferol) in patients with advanced malignancy. AB - Vitamin D is a steroid hormone best known for its activity in regulating calcium and bone metabolism. Epidemiological evidence suggests that vitamin D may play a role in inhibiting the development of colon and prostate cancer. Vitamin D receptors are expressed in many types of malignant cells; in vitro and in vivo vitamin D and vitamin D analogues are active in suppressing the development and inhibiting the growth of numerous human and animal tumors. The major toxicity of the active form of vitamin D, 1,25-dihydroxycholecalciferol (calcitriol), is the induction of hypercalcemia. There are no data indicating the maximum tolerated dose of calcitriol administered every other day (QOD) s.c. We hypothesized that this route and schedule would permit administration of higher doses of calcitriol, which might have anticancer activity. We conducted a Phase I trial of calcitriol given s.c. QOD in patients with advanced solid tumors. Thirty-six patients were entered at doses ranging from 2 to 10 microg QOD; dose-limiting toxicity (hypercalcemia) occurred in three of three patients entered at the 10 microg QOD dose. Hypercalciuria occurred at all dose levels examined. No other toxicity was seen. Assessment of serum calcitriol concentrations by a RIA revealed a decrease in concentration-time curves on day 7 compared to day 1 of therapy. A dose-dependent increase in peak serum level and estimated area under the concentration-time curve was seen. The maximum serum levels occurred at the 10-microg QOD dose: 288 +/- 74 and 321 +/- 36 pg/ml at days 1 and 7, respectively. The normal range of calcitriol serum concentration, determined using this assay, is 16-56 pg/ml. Serum calcitriol levels were maintained at near peak concentrations for at least 8 h following s.c. injection. This study indicates that substantial doses of calcitriol can be administered via this route with tolerable toxicity. Studies to explore approaches to ameliorate the hypercalcemia induced by calcitriol and to explore alternative schedules and interactions with other agents are warranted. PMID- 10389918 TI - High inter- and intrapatient variation in 5-fluorouracil plasma concentrations during a prolonged drug infusion. AB - The purpose of the study was to examine inter- and intrapatient variation in 5 fluorouracil (5-FU) plasma concentrations in adult cancer patients receiving a 3 day drug infusion. Fourteen patients received 1266 mg/m2 N-(phosphonacetyl)-L aspartate (PALA) infused i.v. over 15 min on day 1, followed immediately by a loading dose of 500 mg/m2 calcium leucovorin over 30 min. Then a prolonged infusion of leucovorin at 500 mg/m2/day and 5-FU at 1750 mg/m2/day was administered as either a constant rate or as a circadian infusion over 72 h. During constant rate infusions, 5-FU concentrations within individuals varied by 1.7-fold, but no uniform time of peak or trough concentration was observed. Transformation of these data by setting the time of peak to 0 h and by expressing concentrations as the percentage of the 24-h mean value revealed a nonrandom distribution of the time from peak to trough with a median time of 12 h (P = 0.027). These transformed data were also successfully fit to a circadian cosinor function (P < 0.001). During multiple constant rate 5-FU infusions, the intrapatient variability was high; the times of peak 5-FU concentration occurred at the same approximate sampling time 43% of the time, and troughs coincided 17% of the time. No difference in clinical toxicity was observed when matched constant rate and circadian infusions of 5-FU were compared. High inter- and intrapatient variability exists in 5-FU plasma concentrations in adult cancer patients during constant rate infusions with no evidence of a consistent circadian rhythm in untransformed data. PMID- 10389919 TI - Immunogenicity of granulocyte-macrophage colony-stimulating factor (GM-CSF) products in patients undergoing combination therapy with GM-CSF. AB - In this study, we have assessed the development of neutralizing and nonneutralizing granulocyte-macrophage colony-stimulating factor (GM-CSF) antibodies in two groups of patients with metastatic colorectal carcinoma receiving two different GM-CSF products. Three clinical trials were carried out, and a combination of GM-CSF and a colon carcinoma-reactive antibody was used in the absence of any concomitant chemotherapy. Two different GM-CSF products, both rDNA-derived and produced in Escherichia coli, were used. Patients in Trial 1 received product X, and those in Trials 2 and 3 received product Y. Patients in Trial 2 also received interleukin 2 in an attempt to potentiate immune responses. After the first cycle of treatment, no GM-CSF antibodies were detected, but on subsequent therapy, 28 of the 38 patients tested receiving product Y (Trials 2 and 3) developed antibodies that bound to the GM-CSF product used for therapy. However, none of the patients developed antibodies that neutralized the biological activity of GM-CSF, as assessed using an in vitro bioassay. Furthermore, there was no in vivo impairment in GM-CSF-induced expansion of leukocytes, neutrophils, and eosinophils in the patients. In contrast, 19 of the 20 patients given product X (Trial 1) developed GM-CSF binding antibodies, and 9 of these patients were shown to develop antibodies that neutralized the biological activity of GM-CSF. The presence of the latter was associated with a significant reduction in GM-CSF-induced expansion of leukocytes, neutrophils, and eosinophils in patients. Therefore, product X appears to be more immunogenic than product Y. Immunochemical characterization confirmed that the specificity of the antibody responses varied depending on the product used for therapy. Whereas sera from Trial 1 patients treated with product X showed the presence of antibodies with strong recognition of GM-CSF proteins, sera from patients treated with product Y showed varied recognition of GM-CSF ranging from fairly strong to very weak but bound predominantly to two E. coli-derived, non-GM-CSF-related proteins of Mr approximately 20,000 and Mr approximately 30,000. Therefore, in sera from patients receiving product Y, the antibody specificity appeared to be directed not only against GM-CSF but also against non-product-related host cell contaminants. This study shows that GM-CSF products used for therapy are potentially immunogenic and generate antibodies to GM-CSF and/or other non product-related contaminants. However, only antibodies that neutralize the biological activity of GM-CSF compromise therapeutic efficacy of the cytokine. PMID- 10389920 TI - Phase I study of transforming growth factor-beta3 mouthwashes for prevention of chemotherapy-induced mucositis. AB - The purpose of this study was to establish the safety and tolerability of recombinant transforming growth factor-beta3 (TGF-beta3; CGP 46614) mouthwashes intended for prevention of chemotherapy-induced mucositis. Local effects were especially analyzed by objective and subjective measurements of mucositis. Secondary aims were analysis of potential systemic exposure and development of anti-TGF-beta3-antibodies. Eleven breast cancer patients received chemotherapy with 1.5 g/m2 cyclophosphamide i.v., 80 mg/m2 epirubicin i.v., and 1.0 g/m2 5 fluorouracil i.v. (n = 8) or 1.6 g/m2 carboplatin i.v., 480 mg/m2 thiotepa i.v., and 6 g/m2 cyclophosphamide i.v. divided over 4 days (n = 3). TGF-beta3 mouthwashes (10 ml; provided by Novartis, Basel, Switzerland) were administered for 4 days, four times a day, starting 1 day before chemotherapy. The dose was escalated in following patients from 25 microg/ml (n = 3) to 50 microg/ml (n = 3) and 100 microg/ml (n = 5). Clinically, the mucosa was scored objectively and according to WHO criteria. The percentage of viable oral epithelial cells was determined by trypan blue dye exclusion. Morphology of cells was assessed in buccal smears. Plasma samples were collected for determination of TGF-beta3 levels and anti-TGF-beta3-antibodies. Adverse events were recorded by the patient in a diary. Mouthwashes with TGF-beta3 were well tolerated. Three patients scored for mucositis > grade 0 (WHO grading criteria). The percentage of viable oral epithelial cells in patients treated with 1.5 g/m2 cyclophosphamide i.v., 80 mg/m2 epirubicin i.v., and 1.0 g/m2 5-fluorouracil i.v. was stable, whereas in patients treated with 1.6 g/m2 carboplatin i.v., 480 mg/m2 thiotepa i.v., and 6 g/m2 cyclophosphamide i.v. divided over 4 days, an increase was observed. The morphology of buccal cells showed a transient shift from mature to immature cells in the first week. Neither systemic absorption of TGF-beta3 nor development of TGF-beta3-antibodies was observed. TGF-beta3 mouthwashes were well tolerated and deserve further study in preventing chemotherapy-induced mucositis. PMID- 10389922 TI - Detection of erbB-2 amplifications in tumors and sera from esophageal carcinoma patients. AB - We used TaqMan PCR to detect quantitative anomalies of tumor markers in both tumor and serum DNA from esophageal cancer patients. We demonstrated the potential of this methodology by detecting erbB-2 amplifications in a plurality of esophageal tumor samples. These amplifications were corroborated by Southern blots. We then showed the potential of this methodology to detect quantitative anomalies of erbB-2 in serum DNA from individuals with a corresponding amplification in the tumor. The capability of TaqMan PCR to detect abnormalities in serum of esophageal cancer patients creates an opportunity to diagnose esophageal cancer and to monitor the outcome of treatment with a blood test. PMID- 10389921 TI - Expression of proinflammatory and proangiogenic cytokines in patients with head and neck cancer. AB - Altered immune, inflammatory, and angiogenesis responses are observed in patients with head and neck squamous cell carcinoma (HNSCC), and many of these responses have been linked with aggressive malignant behavior and a decrease in prognosis. In this study, we examined the hypothesis that HNSCC cells produce cytokines that regulate immune, inflammatory, and angiogenesis responses. We identified important regulatory cytokines in supernatants of well-defined and freshly cultured HNSCC cell lines by ELISA and determined whether these cytokines are detected in tumor cell lines and tissue specimens by immunohistochemistry. The serum concentration of the cytokines and cytokine-dependent acute phase inflammatory responses (i.e., fibrinogen, C-reactive protein, and erythrocyte sedimentation rate) from patients with HNSCC was determined, and the potential relationship of serum cytokine levels to tumor volume was analyzed. Cytokines interleukin (IL)-1alpha, IL-6, IL-8, granulocyte-macrophage colony-stimulating factor (GM-CSF), vascular endothelial growth factor (VEGF), and basic fibroblast growth factor were detected in similar concentration ranges in the supernatants of a panel of established University of Michigan squamous cell carcinoma (UM-SCC) cell lines and supernatants of freshly isolated primary HNSCC cultures. Evidence for the expression of IL-1alpha, IL-6, IL-8, granulocyte-macrophage colony stimulating factor, and VEGF in HNSCC cells within tumor specimens in situ was obtained by immunohistochemistry. In a prospective comparison of the cytokine level and cytokine-inducible acute-phase proteins in serum, we report that cytokines IL-6, IL-8, and VEGF were detected at higher concentrations in the serum of patients with HNSCC compared with patients with laryngeal papilloma or age-matched control subjects (at P < 0.05). The serum concentrations of IL-8 and VEGF were found to be weakly correlated with large primary tumor volume (R2 = 0.2 and 0.4, respectively). Elevated IL-1- and IL-6-inducible acute-phase responses were also detected in cancer patients but not in patients with papilloma or control subjects (at P < 0.05). We therefore conclude that cytokines important in proinflammatory and proangiogenic responses are detectable in cell lines, tissue specimens, and serum from patients with HNSCC. These cytokines may increase the pathogenicity of HNSCC and prove useful as biomarkers or targets for therapy. PMID- 10389923 TI - Absence of PTEN germ-line mutations in men with a potential inherited predisposition to prostate cancer. AB - Epidemiological studies have demonstrated that men with a family history of prostate cancer are at an increased risk for this disease. This important observation has led a number of research teams, including our own, to collect DNA samples and clinical data from prostate cancer families, with the goal of localizing and characterizing prostate cancer susceptibility genes. The candidate tumor suppressor gene PTEN (also called MMAC1) has recently been shown to be somatically altered in several common malignancies, including cancers of the brain, kidney, skin, thyroid, endometrium, breast, and prostate. Germ-line mutations in this gene, which maps to chromosome 10q23, have been associated with Cowden disease, an autosomal dominant cancer predisposition syndrome that is characterized by multiple hamartomas. Although prostate cancer is not typically associated with Cowden disease, previous studies of sporadic prostate cancers demonstrate loss of heterozygosity at 10q23 loci in approximately 25% of cases. We, therefore, hypothesized that germ-line mutations in the PTEN gene may predispose to prostate cancer in a subset of families, particularly those in which cancers of the breast, kidney, and/or thyroid also segregate. To test this hypothesis, DNA was isolated from whole blood of 11 prostate cancer patients from 10 unrelated families. Four of the 10 families met the previously established clinical criteria for hereditary prostate cancer. Eight of the II men had at least one second primary malignancy, including cases of neuroendocrine cancer, glioblastoma multiforme, melanoma, kidney, and thyroid cancer. Although we identified some common as well as some unique polymorphisms, no nonsense or missense mutations were identified in any of the 11 samples. To further examine the possibility that PTEN mutations contribute to prostate cancer predisposition, we also studied the probands from each of 10 families with early-onset and/or multiple individuals with prostate cancer. Sequence analysis of the PTEN gene in these 10 men also revealed no mutations or novel polymorphisms. We conclude that germ-line mutations in the PTEN are unlikely to contribute in a significant way to the inherited predisposition to prostate cancer. PMID- 10389925 TI - Rearrangements of chromosome band 1p36 in non-Hodgkin's lymphoma. AB - We studied 850 consecutive cases of histologically ascertained pretreatment non Hodgkin's lymphoma with cytogenetically abnormal clones. The diagnostic karyotypes revealed that 12% of these cases exhibited structural rearrangements involving chromosome band 1p36. Here, we describe the karyotypes of 53 cases containing a 1p36 rearrangement [often involving translocations of unknown material and presented as add(1)(p36)]. We used fluorescence in situ hybridization to determine the origin of the translocation partners. We report three different recurrent translocations involving 1p36. These include der(1)t(1;1)(p36;q21) (three cases), der(1)t(1;1)(p36;q25) (three cases), and der(1)t(1;9)(p36;q13) (four cases). Using cytogenetic and fluorescence in situ hybridization analyses, we have resolved the translocation partners in 31 cases. Rearrangements of band 1p36 were found among different histopathological subtypes. Alterations of 1p36 never occurred as a sole abnormality, and in 42 of 53 cases, alterations of the band 14q32 were observed. The t(14;18)(q32;q21) translocation was present in 35 cases. The significantly high occurrence of 1p36 breakpoint in structural rearrangements and its involvement in recurrent translocations suggest that the region is bearing gene(s) that are important in lymphomagenesis. Our study also showed that cytogenetically evident deletions were frequent in chromosome 1p, almost always involving the p36 region, whereas duplications were rare and never encompassed the p36 region. Chromosome band 1p36 harbors many candidate tumor suppressor genes, and we propose that one or more of these genes might be deleted or functionally disrupted as a molecular consequence of the rearrangements, thus contributing to lymphomagenesis. PMID- 10389926 TI - Enhanced expression of inducible nitric oxide synthase and nitrotyrosine in gastric mucosa of gastric cancer patients. AB - Recent studies (K. Komoto et al., Am. J. Gastroenterol., 93: 1271-1276, 1998) have shown that Helicobacter pylori infection is associated with gastric cancer. However, the mechanism of H. pylori in carcinogenesis has not been clarified. H. pylori infection leads to a sustained production of reactive nitrogen species that may contribute to cause DNA damage. In this study, we examined the expression of inducible nitric oxide synthase (iNOS) and nitrotyrosine in gastric mucosa. The expression of iNOS and nitrotyrosine was examined by immunohistochemistry in 93 patients who initially underwent gastric biopsies between 1975 and 1992. Thirty-four individuals were later found to have gastric cancer at least 2 years after the initial biopsies (group A). The other 59 subjects have shown no evidence of gastric cancer during long-term follow-up. Fifty-one of these patients were positive for H. pylori (group B), and eight were negative for H. pylori (group C). The expression of iNOS and nitrotyrosine in the gastric mucosa was significantly higher in H. pylori-positive groups A and B than in H. pylori-negative group C. Among the H. pylori-positive patients, the expression of iNOS and nitrotyrosine was significantly higher in group A than in group B. These results suggest that high production of iNOS and nitrotyrosine in the gastric mucosa infected with H. pylori may contribute to the carcinogenesis of gastric cancer. PMID- 10389924 TI - Antinuclear antibodies as potential markers of lung cancer. AB - There are multiple case reports of antinuclear antibodies (ANAs) in patients with malignancies, yet to date there has not been a systematic survey of ANAs in lung cancer. We have previously reported that autoantibodies to collagen antigens resembling those found in the connective tissue diseases are consistently detected in the sera from lung cancer patients. In this work, we looked for the presence of ANAs in the sera from these same patients. Sera from 64 patients with lung cancer and 64 subjects without a history of cancer were retrospectively tested for reactivity on immunoblots of nuclear extracts of HeLa, small cell carcinoma, squamous cell carcinoma, adenocarcinoma, large cell carcinoma of the lung, and of normal lung cells. Associations were sought between the reactivities on immunoblots and lung cancer cell type, diagnosis, and progression-free survival by the method of classification and regression trees (CARTs). Cross validated CART analyses indicated that reactivities to certain bands in immunoblots are associated with different types of lung cancer. Some of these autoantibodies were associated with a prolonged survival without disease progression. Our data suggest that autoimmunity is often a prominent feature of lung cancer and that molecular characterization of these antigens may lead to the discovery of proteins with diagnostic and prognostic value. PMID- 10389927 TI - Loss of heterozygosity accumulation in primary breast carcinomas and additionally in corresponding distant metastases is associated with poor outcome. AB - The occurrence of distant metastases is the most feared manifestation of breast cancer, often occurring years after the primary surgery and associated with poor survival. The dominant metastatic clone is characterized by an accumulation of genetic alterations, but it is not actually known at what stage of the metastatic cascade these alterations have occurred. We investigated allelic losses during breast cancer progression in a series of 17 primary breast carcinomas and 22 corresponding brain, liver, lung, and bone metastases (mean metastasis-free interval, 31 months) by analyzing 19 microsatellite markers on seven breast cancer- or metastasis-related chromosomal regions and correlated the incidence of combined loss of heterozygosity (LOH) with metastasis-free and postmetastatic survival. We found that, in comparison with the corresponding primary tumor, additional LOH events are frequently found in metastases and that the incidence of combined LOH in the primary tumor, plus the occurrence of additional LOH events in the distant metastases, correlated significantly with decreased postmetastatic survival. Combined LOH of the three breast cancer-related chromosomal regions alone or in combination with allelic loss at the p53 gene region seems to have a specific influence on the aggressive behavior of metastases. We hypothesize that the occurrence of additional LOH events is either involved in termination of dormancy of micrometastatic tumor cells at distant organ sites or acquired during further progression of metastases. PMID- 10389928 TI - Prognostic value of circulating soluble E-selectin concentrations in node negative breast cancer patients. AB - Several studies have suggested that endothelial cells participate in tumor development. Soluble E-selectin (sE-selectin) is specifically released by activated endothelial cells, and its serum concentration can be considered a marker of endothelial activation. In this study, we assessed the prognostic value of sE-selectin concentrations in node-negative breast cancer patients. Serum sE selectin concentrations were measured by an ELISA method prior to surgery in 456 node-negative breast cancer patients. We analyzed also tumor size (TS), histoprognostic grading, and steroid hormone receptor status. The mean sE selectin concentration was 24.9 +/- 15.0 ng/ml. The sE-selectin concentrations were mildly correlated with the TS but not with the other factors. For prognostic analyses, the median follow-up duration was 7.5 years. The cutoff sE-selectin concentration used was 40 ng/ml. In overall survival studies, univariate analyses demonstrated a prognostic value of sE-selectin, TS, and histoprognostic grading, and multivariate analyses demonstrated a prognostic value of sE-selectin and TS. For disease-free survival, univariate and multivariate analyses demonstrated a prognostic value of sE-selectin and TS. sE-selectin concentration is an easily measurable and strong prognostic factor in node-negative breast cancer patients. These results provide further evidence for the role of adhesion molecules expression by endothelial cells in tumor progression. PMID- 10389929 TI - Chromosomal abnormalities in glioblastoma multiforme by comparative genomic hybridization: correlation with radiation treatment outcome. AB - Glioblastoma multiforme (GM) is the most common and most malignant astrocytoma in adults. After surgery, radiation therapy extends patient survival; however, in vivo response to radiation therapy is variable. The purpose of this investigation was to determine whether the cytogenetic abnormalities of GM differ according to patient response to radiation therapy. Radiation response was defined by either progression [radiation-resistant (RR)] or resolution [radiation-sensitive (RS)] of tumor at the first postradiation radiographic imaging evaluation. Twenty RR and 10 RS frozen tissue specimens were subjected to cytogenetic analysis by comparative genomic hybridization. RS and RR specimens had different cytogenetic aberrations that mapped predominantly to chromosomes 7, 9, 10, 13, and 19. Relative gain of 7 occurred in 70% of the RR and 30% of the RS cases and was the most significant difference involving a single change between the two groups (P = 0.06). RR and RS specimens also differed in their patterns of simultaneous cytogenetic aberrations. A simultaneous gain of chromosomes 7 and 19 was found in 30% of the RR cases but was absent in the RS group. Concurrent loss of 9p23-24 and 13q14 regions was absent in the RS cohort but occurred in 30% of the RR series. This latter cytogenetic pattern was also associated with older age. Amplifications were more common in the RR series, but the difference did not reach statistical significance. The data suggest that GM with different in vivo responses to radiation therapy also differ cytogenetically. PMID- 10389930 TI - Infrequent germ-line mutation of the E-cadherin gene in Japanese familial gastric cancer kindreds. AB - Germ-line mutation of the E-cadherin gene was reported in familial gastric cancer (FGC) kindreds from New Zealand. Therefore, we analyzed all of the exons of E cadherin by PCR-single-strand conformational polymorphism analysis in 16 patients from 14 Japanese FGC kindreds. However, no germ-line mutation was detected, suggesting that a predisposition to FGCs by E-cadherin gene mutation is infrequent in Japanese cases. PMID- 10389931 TI - The prognostic significance of tumor-associated antigen 22-1-1 expression in adenocarcinoma of the uterine cervix. AB - We previously established (K. Sonoda et al., Int. J. Oncol., 6: 1099-1104, 1995) a novel monoclonal antibody, 22-1-1, generated from adenocarcinoma of the uterine cervix, and 22-1-1 antigen (Ag) was expressed in cancer cells derived mainly from the uterus and ovary. In this report, a relationship between 22-1-1 Ag expression and clinicopathological variables and the prognostic significance of 22-1-1 Ag were immunohistochemically investigated in adenocarcinoma of the cervix. Of 56 cases, the 22-1-1 Ag was negative in 7, 1+ in 14, 2+ in 26 and 3+ in 9 instances. The 22-1-1 Ag existed both in the cytoplasm and on the membrane of cancer cells. There was no correlation between 22-1-1 Ag expression and age, stage, grade, myometrial invasion, lymph-vascular space invasion, lymph node metastasis, and parametrial invasion. The estimated 5-year overall survival (OS) of patients with low 22-1-1 Ag expression (-/+) and high 22-1-1 Ag expression (++/ ) were 90.5 and 71.4%, respectively. Patients with high 22-1-1 Ag expression had significantly worse OS than those with low 22-1-1 Ag expression (log-rank test, P = 0.0193). In addition, lymph-node metastasis, age, and clinical stage were significantly related to OS in univariate analysis. Multivariate analysis for OS revealed a prognostic significance in 22-1-1 Ag expression, stage, age, and grade. These data suggest that 22-1-1 Ag expression may be related to prognosis in adenocarcinoma of the cervix. PMID- 10389932 TI - Differential expression of the retinoblastoma gene family members in choroidal melanoma: prognostic significance. AB - We evaluated 55 samples of choroidal melanoma managed by enucleation. Knowing that the immunohistochemical expression of the retinoblastoma gene family members Rb/p105, p107, and pRb2/p130 was inversely correlated with the degree of malignancy in at least some histological types, we investigated the expression of these three proteins in choroidal melanoma. We focused on the relationship between patient survival and the immunohistochemical detection of the retinoblastoma proteins. No correlation with clinical outcome was found for Rb/p105 and p107. However, we found pRb2/p130 to be an independent prognostic factor correlating positively or directly with patient survival times and indirectly or inversely with the degree of malignancy. Demonstration of the prognostic value of the immunohistochemical expression of pRb2/p130 is of significance, even if additional studies are required to confirm these data and to compare the prognostic value of pRb2/p130 immunodetection to that of other recently proposed markers, such as p53. PMID- 10389933 TI - Biological characterization of subgroups of squamous cell lung carcinomas. AB - Recently, Pezzella et al. (Am. J. Pathol., 1997, 151: 1417-1423, 1997) reported on a subgroup of non-small cell lung carcinomas that had no morphological evidence of neoangiogenesis but appeared to grow and were highly aggressive. In this investigation, we subdivided 87 squamous cell lung carcinomas into four subgroups according to angiogenesis (low and high vessel density) and tumor growth (low and high tumor cell proliferation). The aim was to find differences, if any, in the angiogenic status and clinical behavior between these subgroups. We identified a group of tumors with low angiogenesis and high tumor cell proliferation that was characterized by high expression of vascular endothelial growth factor, low expression of basic fibroblast growth factor, reduced apoptosis, increased incidence of metastases, and short survival times. These data show that even squamous cell lung carcinomas are a heterogeneous group of tumors that can be subdivided in tumors with different biological properties and different clinical behaviors. PMID- 10389934 TI - Expression of the 67-kDa laminin receptor in acute myeloid leukemia cells mediates adhesion to laminin and is frequently associated with monocytic differentiation. AB - Lodgement, proliferation, and migration of leukemic cells within bone marrow (BM) microenvironment involves adhesion of these cells to the BM extracellular matrix molecules fibronectin and laminin. The 67-kDa laminin receptor (67LR) is a nonintegrin protein with high affinity for laminin, which plays a critical role in basement membrane invasion and metastasis of cancer cells. By Western blotting, we documented that 67LR was strongly expressed in myelomonocytic THP1 and histiocytic U937 cells and was weakly expressed in promyelocytic HL-60 cells. In HL-60 cells, 67LR expression almost disappeared after retinoic-induced granulocytic differentiation, whereas it strongly increased after phorbol ester induced monocytic differentiation. We did not detect 67LR expression in normal BM hematopoietic cells, in precursor-B acute lymphoblastic leukemia, in chronic lymphocytic leukemia, or in chronic myeloid leukemia in chronic phase. By contrast, we detected enhanced 67LR expression in 40% of 53 de novo acute myeloid leukemias (AMLs), which frequently exhibited monocytic or myelomonocytic morphology and expressed CD14 and CD11a (P < 0.05). Using a colorimetric assay, we found that the expression pattern of this receptor corresponded to a higher adhesion to laminin; the adhesion was specific because in vitro addition to laminin-coated wells of recombinant 37-kDa laminin receptor precursor (37LRP), which is the cytoplasmic precursor containing both laminin-binding domains of cell surface 67LR, significantly reduced laminin binding of AML cells. The expression of 67LR on AML cell surface did not correlate with other differentiation and integrin antigens such as CD7, CD13, CD33, CD34, CD11b, CD11c, CD49d, CD49e, CD45RA, and CD45RO. In contrast with 67LR behavior in solid tumors, no statistically significant difference was found between 67LR expression and any hematological characteristic of the disease at diagnosis, nor between 67LR expression and outcome of the disease as measured by complete remission rate, disease-free survival, or overall survival. In conclusion, our results indicate that 67LR expression mediates specific adhesion to laminin and that the detection of this molecule may be a valuable addition to other lineage-associated antigens in identifying monocytic-oriented AML. PMID- 10389935 TI - Increased serum and urinary levels of a parathyroid hormone-related protein COOH terminus in non-small cell lung cancer patients. AB - Parathyroid hormone-related protein (PTHrP) is expressed in a variety of human cancers including lung cancer. Three mature peptides with different COOH-terminal regions, PTHrP (1-139), PTHrP (1-173), and PTHrP (1-141), are translated from three different mRNAs through alternative splicing. In each, COOH-terminal fragment (C-PTHrP) is stable and measurable in the urine. In the present study, we measured concentrations of circulating and urinary C-PTHrP in 28 patients with primary lung cancer and normal serum calcium levels. We used PCR to evaluate PTHrP mRNA expression and its alternative splicing types in 16 lung cancer cell lines and 17 lung cancer tissues. The average serum C-PTHrP level was 38.95 +/- 19.41 pmol/l in 28 lung cancer patients, whereas that in 10 normal subjects was 26.53 +/- 9.43; the difference was statistically significant (P = 0.0065). Average urine C-PTHrP:urine creatinine ratio was 7.56 +/- 5.17 x 10(-1) pmol/mg creatinine in 28 lung cancer patients, whereas it was 4.91 +/- 1.77 in 10 normal subjects; the difference was statistically significant (P = 0.0287). C-PTHrP radioimmunoassays detected that 23% of non-small cell lung cancer patients had higher serum C-PTHrP levels, and 32% had higher urinary C-PTHrP:urine creatinine ratio than average + 2 SD of normal subjects. Reverse transcription-PCR detected PTHrP mRNA expression in 21 of 21 non-small cell lung cancer (NSCLC) samples and 3 of 12 small cell lung cancer samples. In the cancer cell lines and tissues that had detectable PTHrP mRNA, PTHrP (1-139) mRNA was found in 21 of 24, PTHrP (1 173) mRNA was found in 19 of 24, and PTHrP (1-141) mRNA was found in 23 of 24. Our results suggest that all PTHrP mRNA expression is common in lung cancers. We found that NSCLCs cancers had detectable PTHrP mRNA, and serum and urinary C PTHrP levels in NSCLC patients were significantly higher than those in normal subjects. We concluded that NSCLC produced PTHrP more frequently, but there was no clear significance of C-PTHrP measurement in lung cancer patients for cancer detection using the present assay. We suggested that PTHrP probably plays a role similar to a growth factor or proliferation factor in lung cancer, especially NSCLC, at a level insufficient to cause humoral hypercalcemia of malignancy. PMID- 10389936 TI - Human telomerase reverse transcriptase (hTERT) gene expression in thyroid neoplasms. AB - Ten percent of fine-needle aspirations (FNAs) of the thyroid are deemed "indeterminate" or "suspicious" for malignancy by the cytopathologist, but most of these lesions are benign. Therefore, additional markers of malignancy may prove to be a useful adjunct. The catalytic component of telomerase, human telomerase reverse transcriptase (hTERT), has been found to be reactivated in immortalized cell lines. Reverse transcription-PCR of the hTERT gene revealed expression in 15 (79%) of 19 malignant thyroid neoplasms, including 6 of 6 follicular carcinomas and 9 of 13 papillary carcinomas. In contrast, hTERT gene expression was detected in only 5 (28%) of 18 benign thyroid nodules, including 2 of 7 follicular adenomas and 3 of 11 hyperplastic nodules. All five benign thyroids exhibiting hTERT gene expression had lymphocytic thyroiditis. No normal thyroids exhibited hTERT gene expression. Telomerase enzyme activity was examined in all 37 nodules and was found to correlate with hTERT gene expression in 35 (95%) nodules. The two cases in which telomerase activity and hTERT expression results were discrepant were in two papillary carcinomas that were telomerase activity negative and hTERT positive. Finally, we have demonstrated that hTERT gene expression can be measured in in vivo FNA samples. These results suggest that hTERT expression may be more accurate than telomerase activity in distinguishing benign from malignant and may be measured in FNA samples from suspicious thyroid lesions. PMID- 10389937 TI - High-level expression of EPHB6, EFNB2, and EFNB3 is associated with low tumor stage and high TrkA expression in human neuroblastomas. AB - Neuroblastoma (NB) is a common pediatric tumor of neural crest origin that is biologically and clinically heterogeneous. EPH family receptor tyrosine kinases and ephrin ligands play fundamental roles in neurodevelopmental processes. Recently, we found that NB cell lines expressed several EPHB and EFNB transcripts, which encode EPHB subgroup receptors and ephrin-B subgroup ligands, respectively. To explore the role of EPHB receptors and ephrin-B ligands in the biology of NB, we examined the expression of EPHB and EFNB transcripts in 47 primary NB specimens. Multiple EPHB and EFNB transcripts were expressed in all of the NB tumors examined, suggesting the involvement of these transcripts in modulating the biological behavior of NB. Higher levels of EPHB6, EFNB2, and EFNB3 expression were found in low-stage tumors (stage 1, 2, and 4S) than in advanced-stage tumors (stage 3 and 4; P = 0.0013, P = 0.0048, and P = 0.027, respectively). Expression of TrkA, a well-established prognostic marker of favorable NB, was positively correlated with EPHB6, EFNB2, and EFNB3 expression (P < 0.0001, P = 0.0019, and P = 0.0001, respectively). MYCN-amplified tumors expressed lower levels of EPHB6, EFNB2, EFNB3, and TrkA transcripts compared to nonamplified tumors (P = 0.0006, P = 0.0023, P = 0.0048, and P = 0.0001, respectively). These data suggest that high-level expression of EPHB6, EFNB2, and EFNB3 is associated with favorable NB and that low-level expression of EPHB6, EFNB2, and EFNB3 correlates with aggressive MYCN-amplified NB. Thus, EPHB6, EFNB2, and EFNB3 may have biological relevance in NB. Further investigation on the biology of these genes may help provide insight into the treatment of NB. PMID- 10389938 TI - Messenger RNA determination of estrogen receptor, progesterone receptor, pS2, and plasminogen activator inhibitor-1 by competitive reverse transcription-polymerase chain reaction in human breast cancer. AB - Estrogen receptor (ER), progesterone receptor (PR), the estrogen-inducible protein pS2, and plasminogen activator inhibitor-1 (PAI-1) are important prognostic factors in primary breast cancer. The protein concentrations of these factors in breast tumors have been well documented. However, few data about the mRNA expression of ER, PR, pS2, and PAI-1 in breast cancer are available, which is mostly due to the limitations of conventional techniques for mRNA analysis. We have described a competitive reverse transcription-PCR system for the simultaneous quantification of ER, PR, pS2, and PAI-1 mRNA in tumor samples. Here, we evaluated 100 tumor biopsies from breast cancer patients for the mRNA expression of ER, PR, pS2, and PAI-1. The results were analyzed for correlations with protein status and with clinical data. Significant correlations between mRNA expression levels and protein concentrations of all tested markers were found. In only a few cases was there an obvious discordance between the measurable amounts of mRNA and protein, especially for ER and PR. In addition, ER, PR, and pS2 mRNA levels correlated significantly with each other. No correlation between PAI-1 mRNA amount and the expression of the other markers was found. With respect to clinical data, ER and PR mRNA levels were found to be inversely correlated to tumor size and histological grade but not to the lymph node status. pS2 and PAI-1 mRNA expression were not correlated with tumor size, grade, or lymph node involvement. In conclusion, competitive reverse transcription-PCR may be used as an alternative for the study of prognostic factors in human breast cancer and other malignancies. However, before mRNA expression is measured for diagnostics, a presumed concordance of mRNA and protein expression must be evaluated very carefully for every gene. PMID- 10389939 TI - Decreased expression of retinoic acid receptors, transforming growth factor beta, involucrin, and cornifin in cervical intraepithelial neoplasia. AB - Cervical intraepithelial neoplasia (CIN) I, II, and III represent a spectrum of premalignant epithelial changes and are ideal targets for application of chemoprevention strategies. Intermediate end point biomarkers are increasingly being used as surrogate end points to monitor clinical chemoprevention trials. To identify potential biomarkers in cervical epithelium, we analyzed the expression of nuclear retinoic acid receptor (RAR) mRNA by in situ hybridization, involucrin, cornifin, and transforming growth factors (TGFs) beta1 and beta2 by immunohistochemistry in cervical specimens, which contained adjacent normal epithelium and CIN lesions from 52 patients. These biomarkers were expressed in all adjacent normal cervical epithelia, whereas all CIN lesions including CIN I, CIN II, and CIN III exhibited decreased expression of RAR-alpha by 55.8%, RAR beta by 64.7%, RAR-gamma by 54.9%, involucrin by 80.8%, cornifin by 88.5%, TGF beta1 by 89.7%, and TGF-beta2 by 85.7%. Viewed as a whole, these biomarkers were down-regulated in 100% of the CIN lesions. Because all of these biomarkers can be modulated in vitro by retinoids, they may serve as intermediate biomarkers for retinoid chemoprevention trials in the patients with CIN lesions. PMID- 10389940 TI - Phenylbutyrate induces cell differentiation and modulates Epstein-Barr virus gene expression in Burkitt's lymphoma cells. AB - Although Burkitt's lymphoma (BL) is a readily treated malignancy, recurrences, as well as disease arising in immunosuppressed patients, are notoriously resistant to conventional therapeutic approaches. The EBV is associated with a significant proportion of these lymphomas that evade immune surveillance through decreased expression of both viral and cellular antigens. Increasing the immunogenicity of BL cells may, therefore, represent a potentially beneficial therapeutic maneuver. Using in vitro models of EBV-transformed lymphoblastoid as well as BL cell lines, we demonstrate increased expression of genes coding for HLA class I and EBV latent proteins by the differentiation inducer phenylbutyrate (PB). The aromatic fatty acid also caused cytostasis associated with sustained declines in c-myc expression, a direct antitumor effect that was independent of the EBV status. We conclude, therefore, that differentiation therapy of BL with PB may lead to growth arrest with increased tumor immunogenicity in vivo. The findings may have clinical relevance because the in vitro activity has been observed with PB concentrations that are well tolerated and nonimmunosuppressive in humans, a desirable feature for the different patient populations afflicted with this disease. PMID- 10389941 TI - Ionizing radiation improves survival in mice bearing intracranial high-grade gliomas injected with genetically modified herpes simplex virus. AB - Malignant gliomas remain incurable with current interventions. Encouraging investigational approaches include the use of genetically modified herpes simplex 1 (HSV-1) viruses as direct cytotoxic agents. Combining attenuated HSV-1 with standard therapy, human U-87 malignant glioma xenografts grown in the hind limb or intracranially in athymic nude mice were exposed to ionizing radiation, inoculated with genetically modified HSV R3616, or received both virus and radiation. The combination of virus with fractionated ionizing radiation suggests a synergistic action and results in reduced tumor volumes and longer survivals when compared with treatment with either modality alone. PMID- 10389942 TI - Use of carrier cells to deliver a replication-selective herpes simplex virus-1 mutant for the intraperitoneal therapy of epithelial ovarian cancer. AB - Epithelial ovarian cancer (EOC) remains localized within the peritoneal cavity in a large number of patients, lending itself to i.p. approaches of therapy. In the present study, we investigated the effect of replication-selective herpes simplex virus-1 (HSV-1) used as an oncolytic agent against EOC and the use of human teratocarcinoma PA-1 as carrier cells for i.p. therapy. HSV-1716, a replication competent attenuated strain lacking ICP34.5, caused a direct dose-dependent oncolytic effect on EOC cells in vitro. A single i.p. administration of 5 x 10(6) plaque-forming units resulted in a significant reduction of tumor volume and tumor spread and an increase in survival in a mouse xenograft model. PA-1 cells supported HSV replication in vitro and bound preferentially to human ovarian carcinoma surfaces compared with mesothelial surfaces in vitro and in vivo. In comparison with the administration of HSV-1716 alone, irradiated PA-1 cells, infected at two multiplicities of infection with HSV-1716 and injected i.p. at 5 x 10(6) cells/animal, led to a significant tumor reduction in the two models tested and the significant prolongation of mean survival in one model. Histological evaluation revealed extensive necrosis in tumor areas infected by HSV-1716. Immunohistochemistry against HSV-1 revealed areas of viral infection within tumor nodules, which persisted for several weeks after treatment. Administration of HSV-infected PA-1 carrier cells resulted in larger areas of tumor infected by the virus. Our results indicate that replication-competent attenuated HSV-1 exerts a potent oncolytic effect on EOC, which may be further enhanced by the utilization of a delivery system with carrier cells, based on amplification of the viral load and possibly on preferential binding of carrier cells to tumor surfaces. PMID- 10389943 TI - 125I-labeled anti-epidermal growth factor receptor-vIII single-chain Fv exhibits specific and high-level targeting of glioma xenografts. AB - A single-chain antibody fragment, MR1(scFv), with specific binding to epidermal growth factor receptor-vIII (EGFRvIII), was produced, radiolabeled, and evaluated for biodistribution in human glioma-bearing athymic mice. The mutant receptor EGFRvIII has a deletion in its extracellular domain that results in the formation of a new, tumor-specific antigen found in glioblastomas, breast carcinomas, and other tumors. The scFv molecule, designed as V(H)-(Gly4-Ser)3-V(L), was expressed in Escherichia coli in inclusion body form; recovered scFv fragments were properly refolded in redox-shuffling buffer. Size-exclusion chromatography of purified scFv demonstrated a protein monomer of Mr 26,000. Labeling was performed using N-succinimidyl 5-[125I]iodo-3-pyridinecarboxylate (SIPC) or Iodogen to specific activities of 0.5-2.0 mCi/mg, with yields of 35-50% and 45-70%, respectively. The immunoreactive fraction (IRF) of the labeled MR1(scFv) was 65 80% when SIPC was used and 50-55% when Iodogen was used. The affinity (K(A)) of MRI(scFv) for EGFRvIII was 4.3 x 10(7) +/- 0.1 x 10(7) M(-1) by BIAcore analysis, and it was 1.0 x 10(8) +/- 0.1 x 10(8) M(-1) and by Scatchard analysis versus EGFRvIII-expressing cells. After incubation at 37 degrees C for 24 h, the binding affinity was maintained, and the IRF was maintained at 60-70%. The specificity of MR1(scFv) for EGFRvIII was demonstrated in vitro by incubation of radiolabeled MR1(scFv) with the EGFRvIII-expressing U87MG.deltaEGFR cell line in the presence or absence of competing unlabeled MR1(scFv) or anti-EGFRvIII MAbs L8A4 and H10. In biodistribution studies using athymic mice bearing s.c. U87MG.deltaEGFR tumor xenografts, animals received intratumoral or i.v. infusions of paired-label [125I]SIPC-MR1(scFv) and [131I]SIPC-anti-Tac(scFv) as a control. When given by the intratumoral route, MR1(scFv) retained high tumor uptakes of 85% injected dose per gram of tissue at 1 h and 16% injected dose per gram of tissue at 24 h following administration. Specific: control scFv tumor uptake ratios of more than 20:1 at 24 h demonstrated specific localization of MR1(scFv). The excellent tumor retention of MR1(scFv), combined with its rapid clearance from normal tissues, resulted in high tumor:normal organ ratios. PMID- 10389944 TI - Combination surgery and nonviral interleukin 2 gene therapy for head and neck cancer. AB - We have developed a novel nonviral interleukin 2 (IL-2) gene therapy that demonstrates significant treatment-specific, antitumor efficacy in combination with subtotal surgical resection in a head and neck cancer murine model. Treatment of established head and neck tumors in immunocompetent mice was performed via direct injection with a cationic liposome composed of DOTMA and cholesterol formulation carrying DNA plasmid for human IL-2 (hIL-2) gene expression. ELISA assays of tumor extracts 24 h after treatment of hIL-2 gene therapy revealed increased local hIL-2 production as well as a formulation specific secondary induction of murine IFN-gamma and IL-12. We hypothesize that the paracrine production of multiple cytokines after IL-2 single gene transfer is important for generating a therapeutic effect, and that this strategy will be well tolerated and effective in combination with surgery for head and neck cancer. In animal experiments where surgery was performed in conjunction with an operative site injection of hIL-2 plasmid formulation, no pre-, intra-, or postoperative toxicity or compromise to wound healing was identified. In murine experiments combining partial surgical resection with the nonviral gene therapy, significant antitumor efficacy was demonstrated in the hIL-2 plasmid formulation group compared with empty plasmid formulation and lactose-injected controls. In a separate experiment using smaller tumor sizes, we also demonstrated that treatment outcomes were dependent on the technical aspect of the actual treatment injection as well as visualization with surgical access. The hIL-2 plasmid formulation gene therapy induces local expression of multiple cytokines, results in treatment-specific antitumor effects, and circumvents many of the concerns and toxicity encountered with viral gene transfer. These data support the need for continued preclinical investigation and the consideration of human clinical trials for combination nonviral hIL-2 gene therapy and surgery for head and neck cancer. PMID- 10389945 TI - Purging of contaminating breast cancer cells from hematopoietic stem cell grafts by adenoviral GAL-TEK gene therapy and magnetic antibody cell separation. AB - The presence of contaminating tumor cells in autologous bone marrow or peripheral blood stem cell (PB-SC) preparations increase the likelihood of relapse in women receiving transplants for metastatic breast cancer. We describe a new technique for purging breast cancer cells (BCCs) that combines two independent strategies: (a) the specific enrichment of CD34+ progenitor stem cells by magnetic antibody cell separation (MACS), and then (b) infection of the contaminating BCCs with a recombinant adGAL-TEK marker/suicide gene adenovirus (ad-v), followed by the addition of ganciclovir (GCV). Infection with this ad-v results in three to four times greater expression of ad-v-delivered reporter gene in BCCs than in CD34+ cells. In addition -2 h, -low multiplicity of infection (50:1) adGAL-TEK infections of BCC lines (MCF-7 and BT474) eradicated >99% of BCCs after 72 h of exposure to 20 microM GCV. However, exposure to both adenovirus and GCV at the MOIs and doses used had little effect on hematopoietic stem cells to form colonies in colony-forming unit assays. adGAL-TEK infection in our model system (10(3)-10(5) BCCs added into 10(7) HSCs) also resulted in the 3 to 5 log eradication of clonogenic BCCs after the addition of GCV. MACS enrichment/purification of CD34+ cells from PB-SC contaminated with 2 x 10(6) to 5 x 10(7) BCCs followed by adGAL-TEK infection and GCV addition resulted in 5-7 log depletion of clonogenic BCCs as well as enrichment of CD34+ progenitor cells to >98%, with the recovery of >70% of hematopoietic stem cells. This adenoviral purging system is so robust that poor MACS purification, resulting in 1.5-log depletion of BCCs, still permits excellent ad-v infection and BCC killing. PMID- 10389946 TI - Structure-based design of specific inhibitors of Janus kinase 3 as apoptosis inducing antileukemic agents. AB - A novel homology model of the kinase domain of Janus kinase (JAK) 3 was used for the structure-based design of dimethoxyquinazoline compounds with potent and specific inhibitory activity against JAK3. The active site of JAK3 in this homology model measures roughly 8 A x 11 A x 20 A, with a volume of approximately 530 A3 available for inhibitor binding. Modeling studies indicated that 4 (phenyl)-amino-6,7-dimethoxyquinazoline (parent compound WHI-258) would likely fit into the catalytic site of JAK3 and that derivatives of this compound that contain an OH group at the 4' position of the phenyl ring would more strongly bind to JAK3 because of added interactions with Asp-967, a key residue in the catalytic site of JAK3. These predictions were consistent with docking studies indicating that compounds containing a 4'-OH group, WHI-P131 [4-(4' hydroxyphenyl)-amino-6,7-dimethoxyquinazoline], WHI-P154 [4-(3'-bromo-4' hydroxylphenyl)-amino-6,7-dimethoxyquinazoline], and WHI-P97 [4-(3',5'-dibromo-4' hydroxylphenyl)-amino-6,7-dimethoxyquinazolin e], were likely to bind favorably to JAK3, with estimated K(i)s ranging from 0.6 to 2.3 microM. These compounds inhibited JAK3 in immune complex kinase assays in a dose-dependent fashion. In contrast, compounds lacking the 4'-OH group, WHI-P79 [4-(3'-bromophenyl)-amino 6,7-dimethoxyquinazoline], WHI-P111 [4-(3'-bromo-4'-methylphenyl)-amino-6,7 dimethoxyquinazoline], WHI-P112 [4-(2',5'-dibromophenyl)-amino-6,7 dimethoxyquinazoline], WHI-P132 [4-(2'-hydroxylphenyl)-amino-6,7 dimethoxyquinazoline], and WHI-P258 [4-(phenyl)-amino-6,7-dimethoxyquinazoline], were predicted to bind less strongly, with estimated K(i)s ranging from 28 to 72 microM. These compounds did not show any significant JAK3 inhibition in kinase assays. Furthermore, the lead dimethoxyquinazoline compound, WHI-P131, which showed potent JAK3-inhibitory activity (IC50 of 78 microM), did not inhibit JAK1 and JAK2, the ZAP/SYK family tyrosine kinase SYK, the TEC family tyrosine kinase BTK, the SRC family tyrosine kinase LYN, or the receptor family tyrosine kinase insulin receptor kinase, even at concentrations as high as 350 microM. WHI-P131 induced apoptosis in JAK3-expressing human leukemia cell lines NALM-6 and LC1;19 but not in melanoma (M24-MET) or squamous carcinoma (SQ20B) cells. Leukemia cells were not killed by dimethoxyquinazoline compounds that were inactive against JAK3. WHI-P131 inhibited the clonogenic growth of JAK3-positive leukemia cell lines DAUDI, RAMOS, LC1;19, NALM-6, MOLT-3, and HL-60 (but not JAK3-negative BT 20 breast cancer, M24-MET melanoma, or SQ20B squamous carcinoma cell lines) in a concentration-dependent fashion. Potent and specific inhibitors of JAK3 such as WHI-P131 may provide the basis for the design of new treatment strategies against acute lymphoblastic leukemia, the most common form of childhood cancer. PMID- 10389947 TI - Penetration of anticancer drugs through solid tissue: a factor that limits the effectiveness of chemotherapy for solid tumors. AB - Penetration of anticancer agents to cells distant from the vascular system is required for efficacy of cancer chemotherapy against solid tumors. Many solid tumors have a poorly formed blood vascular system with variable rates of blood flow and much larger intercapillary distances than those found in normal tissues. The requirement for drugs to penetrate several layers of tissue might pose a barrier to the effective treatment of solid tumors. Multicellular layers (approximately 200 microm thick) were grown in vitro on Teflon membranes from EMT6 murine and MCF7 human tumors and have been used to quantitate the penetration of four widely used anticancer drugs through solid tissue. The penetration of doxorubicin and mitoxantrone was limited and very slow (<10% of the rate of penetration through the Teflon membrane alone). The penetration of methotrexate and 5-FU was more rapid (approximately 30-50% of the rate of penetration through the Teflon membrane alone), but remains a substantial barrier to the effectiveness of these drugs. Strategies to improve the penetration of anticancer drugs through poorly vascularized tumor tissue have considerable potential to improve the outcome of chemotherapy for solid tumors. PMID- 10389948 TI - Vitronectin, a glioma-derived extracellular matrix protein, protects tumor cells from apoptotic death. AB - Vitronectin (VN) is an extracellular matrix (ECM) protein, the synthesis of which in vivo by glioma cells correlates with tumor grade. Although the role of VN as a permissive substrate for glioma migration has been well characterized, its role in conferring a survival advantage for tumor cells has not been addressed previously. By using an in vitro assay of DNA fragmentation as a quantitative measure of apoptotic cell death, we sought to determine whether the sensitivity of two human glioma cell lines (D54 and U251) to drug-induced apoptosis could be inhibited by VN. As well, the extent to which apoptosis could be inhibited was correlated with the levels of the Bcl-2 family of proteins that are known to modulate apoptosis and chemoresistance. Results of the study were: (a) VN coatings, in a dose-dependent manner, inhibited topoisomerase (Topo)-induced apoptosis by up to 50% (optimal coating density, 500 ng/cm2); in contrast, fibronectin (FN), an ECM protein present in abundance in the brain, demonstrated no protection; (b) in a dose-response study, VN clearly conferred a survival advantage (LD50 of Topo: on VN, 120 ng/ml; on FN, 35 ng/ml); (c) the protective effect of VN was not due to enhanced cell adhesion or alterations in the cell cycle distribution; (d) both of the classic integrin receptors that bind VN (alpha(v)beta3, alpha(v)beta5) were capable of mediating this protective effect, because ligation of either of the two classic integrins conferred chemoresistance to Topo; and (e) chemoresistance observed with VN was associated with an increase in expression of two antiapoptotic proteins, Bcl-2 and Bcl-X(L), with a consequent increase in the ratios for Bcl-2:Bax and Bcl-X(L):Bax. VN, an ECM protein preferentially expressed at the tumor-brain interface in vivo, may confer a survival advantage to glioma cells at the advancing tumor margin and may thus, in part, underlie the high level of tumor recurrence at this interface. PMID- 10389949 TI - CLAR1, a novel gene that exhibits enhanced expression in advanced human prostate cancer. AB - The molecular events involved in prostate cancer progression are, at present, poorly understood. Using a differential display technique, we identified a cDNA fragment that is present in greater abundance in stage D prostate tumors compared to stage B tumors. Northern analysis was used to confirm that transcripts for this gene are expressed at higher levels in prostate tumors of later pathological stage and higher Gleason grade compared to tumors of earlier stage and lower grade. These transcripts were also expressed at high levels in all four human prostate cancer cell lines, the neonatal prostate cell line FNC 267beta1, and in a variety of other normal human adult and fetal tissues. The cDNA fragment obtained by differential display was used as a probe to clone the full-length cDNA for this gene from a human heart cDNA library. DNA sequence analysis confirmed that the cDNA was novel, and we have named this gene CLAR1. The gene displays two transcripts of 2.6 and 2.0 kb in all tissues examined. CLAR1 maps to chromosome 19q13.3 and appears highly conserved among mammals. The deduced amino acid sequence of CLAR1 encodes a proline-rich protein that contains several SH3 binding domains and a serine phosphorylation site. The presence of these motifs suggests a possible role for CLAR1 in one or more signal transduction pathways. The enhanced expression of this novel gene in more advanced forms of prostate cancer and its potential role in signal transduction both argue that this gene should be further investigated. PMID- 10389950 TI - Intravenous midazolam for sedation of children undergoing procedures: an analysis of age- and procedure-related factors. AB - OBJECTIVE: This study was performed to determine the doses of midazolam used for sedation during procedures in children, and the frequency of adverse events. METHODS: We performed a retrospective analysis of data collected for a prospective study of flumazenil in children who had received midazolam for a procedure (n = 91, 1-17 years). RESULTS: Practitioners used a wide range of total midazolam doses (0.03-0.6 mg/kg); mean doses ranged from 0.09 +/- 0.06 mg/kg in adolescents to 0.26 +/- 0.13 mg/kg in toddlers (P < 0.001). Opioids were also used in 84% of patients. Twenty-six percent of children with normal lungs, most of whom had received relatively high opioid doses, developed decreased oxygen saturation (as low as 65%) after sedation. Other adverse events included airway obstruction (n = 3) and vomiting (n = 1). CONCLUSIONS: The frequent choice of midazolam, usually combined with an opioid, indicates its wide acceptance. Midazolam doses were inversely related to age. The presence of vomiting, airway obstruction, and decreased oxygen saturation underlines the importance of appropriate personnel, equipment, and monitors during sedation. PMID- 10389951 TI - End-tidal carbon dioxide monitoring during sedation with a combination of midazolam and ketamine for children undergoing painful, invasive procedures. AB - BACKGROUND: Previous studies evaluating the respiratory effects of sedation regimens have focused on events such as a decline in O2 saturation or apnea. The current study used both end-tidal carbon dioxide (ETCO2) monitoring and pulse oximetry to evaluate the respiratory effects of midazolam and ketamine. METHODS: Fifty children who required sedation during invasive procedures formed the cohort for the study. During sedation, ETCO2 was sampled from nasal cannulae of spontaneously breathing patients and measured by a side-stream aspirating infrared device. RESULTS: During the procedure, O2 saturation decreased by 3% or more in three patients. Supplemental oxygen at 2 liters per minute was administered to these patients. The lowest oxygen saturation was 84%. During the total of 767 minutes of monitoring, there were 3068 ETCO2 values recorded. The high ETCO2 values ranged from 37 to 53 mmHg (40.5 +/- 3.3 mmHg). Ninety percent, or 2760, of the values were 40 mmHg or less, 7% or 214 were between 41 and 45 mmHg, 3% or 92 were between 46 and 49 mmHg, and 2 isolated values were greater than 50 mmHg. One episode of airway obstruction was identified by noting cessation of the ETCO2 waveform. This was relieved by repositioning the patient's airway. The three episodes of O2 desaturation, two ETCO2 values greater than 50 mmHg, and the episode of upper airway obstruction all occurred in three patients. Two of these patients had trisomy 21 with macroglossia, and the third had had a recent upper respiratory infection and a history of tonsillar hypertrophy. CONCLUSION: The incidence of adverse cardiorespiratory events associated with the current sedation regimen of midazolam-ketamine is lower than that reported with other commonly used regimens. The addition of ETCO2 monitoring provides an additional monitor to allow for early detection of airway obstruction or subclinical degrees of respiratory depression. PMID- 10389952 TI - Seat belt use by children brought to an inner-city hospital. AB - OBJECTIVE: To characterize restraint use among children brought to an inner-city hospital by private car or taxicab. DESIGN: Cross-sectional survey and direct observation of a convenience sample. SETTING: Main entrance and clinic entrance of a large urban public hospital. PARTICIPANTS: Direct observation was made on 352 children brought by 257 vehicles. One hundred seventy-seven parents or caretakers responded to questionnaires for 240 children. INTERVENTION: None. MEASURES AND MAIN RESULTS: Of the 352 children who were directly observed, 256 (73%) arrived by taxicabs and 96 (27%) by private cars. Thirty-three of 352 (9%) children were observed to be appropriately restrained. Children brought by taxicabs were significantly less likely to be restrained than children brought by private cars (1% vs 31%, P < 0.001). Caretakers reported that seat belts were available in 46 of 54 (85%) private cars, compared to 38 of 88 (43%) taxicabs (P < 0.01). Twenty percent of caretakers who came by taxicabs did not check for seat belts. CONCLUSION: Taxicabs, which are exempt from the New York State's mandatory seat belt law, are a common mode of transportation for children in the inner city. While the overall use of child restraints in the study sample is low, it is particularly low for children in taxicabs. The low rate may be related to both the decreased availability of seat belts and the lack of the mandatory seat belt law for taxicabs. Strategies should be sought to improve child restraint availability in taxicabs and mandate seat belt use. PMID- 10389954 TI - Determination of systolic blood pressure via pulse oximeter in transported pediatric patients. AB - OBJECTIVE: To compare pulse oximetry waveform systolic blood pressure measurements (POWSBP) to measurements obtained by noninvasive blood pressure measurement (NIBPM) during the transport of children. DESIGN: A prospective, convenience sample. SETTING AND PARTICIPANTS: All patients transported by a dedicated Pediatric Critical Care Transport Team were eligible for inclusion. Senior transport nurses with over 3 years of transport experience who had been instructed by the principal investigator (PI) in the technique, obtained the measurements and recorded the results. A convenience sample was obtained based on the presence or absence of one of the senior nurses on the transport team. METHODS: Measurements of blood pressure were obtained by POWSBP and NIBPM (PROPAQ@; Protocol Systems, Beaverton, OR) on pediatric patients during transport in moving ambulances or fixed wing aircraft. Measurement of systolic blood pressure by pulse oximetry (POWSBP) was obtained on all patients by observing the return of the plethysmographic waveform of the pulse oximeter as the blood pressure cuff deflated. The patients were divided into two groups; in group A POWSBP measurements were obtained by using the automated BP cuff from the PROPAQ@ and in group B by a manual BP cuff. These measurements were compared to NIBPM readings obtained at the same time. Between 3 and 13 paired readings were obtained for each patient. The actual transport time determined the number of measurements obtained. RESULTS: A total of 24 patients were enrolled in the study. Multiple matched pair readings (both POWSBP and NIBPM) were obtained from each patient for a total of 180 data points. A two variable linear regression model was run which identified a significant correlation between POWSBP and NIBPM. Group A, r = 0.7592 and r = 0.9477 for group B significant at P< 0.000001. Further, a corrective equation was developed for use with an automated BP cuff. CONCLUSIONS: The use of pulse oximetry waveform systolic blood pressure measurement is a quick and easy method with which to obtain systolic blood pressure in children during transport and shows a close correlation to the standard noninvasive blood pressure measurement. If an automated blood pressure cuff is used, then a corrective equation [NIBP-S = 41.686 + 0.7377(POBPS)] is required. Further validation in a larger group of patients is recommended. PMID- 10389953 TI - Effect of ketorolac in pediatric sickle cell vaso-occlusive pain crisis. AB - BACKGROUND: Ketorolac is a parenteral, nonsteroidal analgesic that does not have a narcotic's risks of respiratory depression, hypotension, or dependence. Its usefulness in providing pain relief in pediatric patients with acute vaso occlusive crisis of sickle cell disease has not been studied to date. METHODS: Twenty-nine patients with sickle cell disease between the ages of 5 and 18 years who presented to The Children's Hospital of Alabama emergency department (ED) with 41 distinct episodes of acute vaso-occlusive pain crisis were enrolled prospectively and randomized to receive either 0.9 mg/kg intravenous (IV) ketorolac or placebo in a double-blind fashion. All patients also received IV fluids and an initial 0.1 mg/kg of IV morphine. Subsequent standardized doses of morphine were given every 2 hours over a 6-hour observation period based upon severity of pain as scored by a 10-cm linear visual analog scale (VAS). Vital signs and pain severity were recorded initially and assessed hourly. Disposition was made at the end of the observation period. RESULTS: Patients receiving ketorolac and those receiving placebo were of similar age, weight, gender, number of prior ED visits, number of prior hospital admissions, duration of pain prior to presentation, and initial pain score. The total dose of morphine received, reduction in severity of pain as measured by VAS, rate of hospital admission, and rate of return to the ED for discharged patients did not differ significantly between the two groups. CONCLUSION: We were unable to demonstrate a synergistic analgesic effect for ketorolac in the treatment of pain from acute vaso-occlusive crisis in pediatric sickle cell disease. Further investigations involving larger samples of sickle cell patients may be needed to further define a role for ketorolac in the acute management of sickle cell vaso-occlusive pain. PMID- 10389955 TI - Purpura fulminans associated with Streptococcus pneumoniae infection in a child. AB - BACKGROUND: Neisseria meningitidis is the most frequent isolate associated with purpura fulminans in children. Although Streptococcus pneumoniae infection has been associated with purpura fulminans, with the exception of one adult, it has only been reported in immunocompromised hosts. PURPOSE: We report an apparently previously healthy child who presented with purpura fulminans associated with pneumococcal meningitis. METHODS: Case report and review of the medical literature from September 1966 to June 1997, using a MEDLINE search. CONCLUSION: While systemic pneumococcal infection is common in childhood, progression to purpura fulminans does not typically occur in overtly healthy children. Our patient illustrates that invasive pneumococcal infection should be considered and empirically treated in a child who presents with purpura fulminans, even in the absence of preexisting functional or anatomic asplenia. PMID- 10389956 TI - Ear-piercing techniques as a cause of auricular chondritis. AB - OBJECTIVE: To investigate the different methods of ear and body piercing as possible sources of infection, and to provide a brief literature review of infections resulting from high ear piercing as well as bacterial coverage of common disinfectants used as preparation agents. METHODS: Two cases of auricular chondritis caused by Pseudomonas are presented. A survey of 14 businesses that pierce ears was conducted using a scheduled interview. Information regarding the type of piercing instrument, composition of earring, training of employees, anatomic placement of earrings, and preparation and aftercare of ears was obtained. RESULTS: One hundred percent of the interviews attempted were completed. The cosmetic shops and earring kiosks both used hand-powered earring "guns" to pierce ears, while the tattoo parlors used sterile needles and forceps. All of the businesses interviewed used earrings composed of either 14K or 24K gold, stainless steel, and other piercing-grade metals. None of the businesses used earrings made of nickel. The cosmetic shops and kiosks used a combination of videos, demonstrations, and direct supervision to train employees but did not have a specified training period. The tattoo parlors required their employees to complete an apprenticeship training program of varying time lengths. All of the businesses pierced the lobe and cartilaginous portions of the ear. The cosmetic shops and kiosks used benzalkonium chloride or isopropyl alcohol as ear preparation agents, while the tattoo parlors used only iodine-based solutions. At all of the businesses, minimal aftercare instructions were given and they typically dealt with maintaining ear-hole patency. CONCLUSION: The cosmetic shops and earring kiosks used piercing methods that predisposed to auricular chondritis, such as poor training of employees and use of benzalkonium chloride as a preparation agent. Emergency physicians need to be aware of the severity of these types of infections, which often require surgical management and intravenous antibiotics covering Pseudomonas. PMID- 10389957 TI - Use of naloxone to reverse symptomatic tetrahydrozoline overdose in a child. PMID- 10389958 TI - Apparent life-threatening events presenting to a pediatric emergency department. AB - OBJECTIVE: To review the etiology, clinical decision-making process, and outcomes of apparent life-threatening events (ALTEs) presenting to a children's hospital emergency department (ED). DESIGN: Retrospective patient record review. SUBJECTS: One hundred thirty infants under the age of 1 year fulfilling the diagnostic definition of an apparent life-threatening event. RESULTS: In a calendar year, 130 infants presented to a large children's hospital ED. The total number of ALTEs studied was 196. The median age was 2 months, and 50% of infants had a normal clinical examination. Eighty-three percent of ALTEs resulted in admission to the hospital. The approach to investigation and management of an ALTE during admission appeared unstructured. Discharge diagnoses, both from the ED and the inpatient service, were numerous, the most common being convulsion, febrile convulsion, GOR, and lower respiratory tract infection. The diagnosis frequently changed in those attending more than once. Eighteen months after cessation of data collection, no infants had died. Follow-up information revealed a higher than-expected prevalence of asthma and seizures. CONCLUSIONS: This is a diverse group of infants, many of whom appear normal following the ALTE. There are many possible diagnoses, but diagnosis correlates poorly with presenting symptoms. It also appears that many commonly performed investigations conducted in this group of infants may not be those that are most helpful for diagnosis, and doctors may be making diagnoses with little supportive evidence. Until research on this group of "first-presentation" infants provides management guidelines for family and emergency doctors, it may be prudent to advise that all such infants presenting with an ALTE should be admitted for a period of observation and further investigation. This would help ensure more accurate diagnosis, as well as provide reassurance for the family. PMID- 10389959 TI - Pediatric chest pain induced by tetracycline ingestion. PMID- 10389960 TI - Amitriptyline-associated seizures in a toddler with Munchausen-by-proxy. AB - We describe an unusual case of a toddler diagnosed with an idiopathic seizure disorder that later was proved to be caused by deliberate administration of amitriptyline by his custodian. In spite of seizures associated with widened electrocardiographic wave (QRS) and right axis deviation on the electrocardiogram (EKG), the correct diagnosis eluded clinicians through a series of hospital admissions. Unfortunately, clinicians are quite accustomed to the fact that patients previously diagnosed with epilepsy have seizures and may not investigate other causes of seizure. This allowed classic signs of cyclic antidepressant poisoning to go unrecognized. PMID- 10389961 TI - Management of children with acute asthma in the emergency department. PMID- 10389962 TI - ABCs of scoring systems for pediatric trauma. AB - This review presents an overview of scoring systems used in pediatric and adult trauma. Triage scoring systems, using readily available physical examination, physiologic, and/or mechanism of injury parameters, are used to determine appropriate prehospital referral patterns. The Trauma Score, Revised Trauma Score, Circulation/Respiration/Abdomen/Motor/Speech Scale, Prehospital Index, and Trauma Triage Rule were reviewed. Injury scoring systems based upon anatomic descriptions of all identified injuries, are retrospectively used to analyze trauma populations. The Abbreviated Injury Scale, Injury Severity Score, Modified Injury Severity Score, Organ Injury Scaling, and Anatomic Profile were discussed. The two trauma outcome analysis systems presented, TRISS and ASCOT, allow for reproducible quantification of trauma severity, and survival comparison between trauma populations. Many of these triage, injury severity, and outcome analysis systems were developed with patient survival as the major outcome variable. Although subsequent studies may have found them to have some predictive value for measures of trauma morbidity, these scoring systems do not specifically address long-term risk of impairment, and therefore overlook one of the most crucial elements of pediatric trauma care. The last 2 decades have seen considerable development of scoring systems and analysis methods applicable to the trauma patient. As presented, this trend includes both the elaboration of increasingly simple, field-oriented triage tools, and more complex mathematical techniques for trauma outcome analysis. Although not all systems were designed specifically with the pediatric patient in mind, validation or modification of these systems for the pediatric patient will likely occur in the future. It is anticipated that this field will continue to evolve with greater mathematical sophistication; a baseline familiarity of the early stages of this evolution may be of benefit to those caring for the pediatric trauma patient. PMID- 10389963 TI - Needle cricothyroidotomy revisited. AB - Needle cricothyroidotomy may provide a life-saving airway when tracheal intubation is not possible. Indications for needle cricothyroidotomy are discussed. Methods of needle/angiocatheter insertion and proposed means to connect to an oxygen source for intermittent insufflation are reviewed. A new technique for administering effective oxygenation and ventilation through a trans tracheal catheter using materials commonly available in an emergency department is presented. Potential complications are discussed. Complete upper airway obstruction is a contraindication to needle cricothyroidotomy because of the risks of barotrauma. In a crisis situation, the emergency practitioner needs a simple, reliable, effective, and preplanned technique to deal with the "nightmare airway." PMID- 10389964 TI - Management of sulfonylurea ingestions. AB - In the majority of pediatric patients with an unintentional ingestion of a sulfonylurea, observation and, if necessary, intravenous glucose supplementation, are sufficient. However, with cases of persistent hypoglycemia or cases refractory to IV glucose supplementation, attempts to inhibit insulin secretion should be considered. Octreotide appears effective and safe. It may eliminate the need for prolonged infusions of hypertonic dextrose solutions and secondarily the risks associated with central line access. It has been successful in restoring euglycemia in cases refractory to glucose infusions. In the absence of octreotide, diazoxide remains a viable alternative to decrease insulin secretion. PMID- 10389965 TI - Pediatric emergency medicine: legal briefs. PMID- 10389966 TI - Diplopia after head injury. PMID- 10389967 TI - Aggressiveness of care. PMID- 10389968 TI - Use of topical lidocaine in pediatric laceration repair. PMID- 10389970 TI - Exclusion of the genes CDKN2 and PTEN as causative gene defects in Li-Fraumeni syndrome. AB - We have analysed Li-Fraumeni syndrome families, previously shown to be negative for mutations in TP53, for mutations to the tumour suppressor genes PTEN and CDKN2. These genes function in cell cycle progression or are mutated in a variety of tumours. We have detected no mutations in the family members tested. PMID- 10389969 TI - Retinoids: present role and future potential. AB - Vitamin A and its biologically active derivatives, retinal and retinoic acid (RA), together with a large repertoire of synthetic analogues are collectively referred to as retinoids. Naturally occurring retinoids regulate the growth and differentiation of a wide variety of cell types and play a crucial role in the physiology of vision and as morphogenic agents during embryonic development. Retinoids and their analogues have been evaluated as chemoprevention agents, and also in the management of acute promyelocytic leukaemia. Retinoids exert most of their effects by binding to specific receptors and modulating gene expression. The development of new active retinoids and the identification of two distinct families of retinoid receptors has led to an increased understanding of the cellular effects of activation of these receptors. In this article we review the use of retinoids in chemoprevention strategies, discuss the cellular consequences of activated retinoid receptors, and speculate on how our increasing understanding of retinoid-induced signalling pathways may contribute to future therapeutic strategies in the management of malignant disease. PMID- 10389971 TI - Microsatellite instability and mismatch repair gene inactivation in sporadic pancreatic and colon tumours. AB - Genomic instability has been proposed as a new mechanism of carcinogenesis involved in hereditary non-polyposis colorectal cancer (HNPCC) and in a large number of sporadic cancers like pancreatic and colon tumours. Mutations in human mismatch repair genes have been found in HNPCC patients, but their involvement in sporadic cancer has not been clarified yet. In this study we screened 21 pancreatic and 23 colorectal sporadic cancers for microsatellite instability by ten and six different microsatellite markers respectively. Microsatellite alterations were observed at one or more loci in 66.6% (14/21) of pancreatic cancers and in 26% (6/23) colon tumours, but all the pancreatic and half of the colon samples showed a low rate of microsatellite instability. All the unstable samples were further analysed for mutations in the hMLH1 and hMSH2 genes and for hypermethylation of the hMLH1 promoter region. Alterations in the hMLH1 gene were found only in colorectal tumours with a large presence of microsatellite instability. None of the pancreatic tumours showed any alteration in the two genes analysed. Our results demonstrate that microsatellite instability is unlikely to play a role in the tumorigenesis of sporadic pancreatic cancers and confirm the presence of mismatch repair gene alterations only in sporadic colon tumours with a highly unstable phenotype. PMID- 10389972 TI - A comparative study of the effects of genistein and 2-methoxyestradiol on the proteolytic balance and tumour cell proliferation. AB - The cytotoxicity of two compounds described as anti-angiogenic, the isoflavone genistein and the oestrogen metabolite 2-methoxyestradiol, has been studied in different human tumour cell lines. Since the degradation of the extracellular matrix is one of the essential steps in angiogenesis, the potential modulatory effects of both compounds on the proteolytic balance in media conditioned by different human tumour cells have been also investigated. The IC50 values for 2 methoxyestradiol were lower than those for genistein on all the cell lines tested. In all the cell lines expressing measurable amounts of active enzymes, genistein induced a shift towards antiproteolysis in both matrix metalloproteinase/tissue inhibitor of metalloproteinase and urokinase/plasminogen activator inhibitor proteolytic balances. On the other hand, 2-methoxyestradiol did not produce any clear net shift of the proteolytic balance, with the significant exception of the matrix metalloproteinase/tissue inhibitor of metalloproteinase balance in WAC-2 cells, a neuroblastoma cell line with enhanced expression of the N-myc oncogene. PMID- 10389973 TI - Generation of reactive oxygen species by human mesothelioma cells. AB - Malignant mesothelioma cells contain elevated levels of manganese superoxide dismutase (MnSOD) and are highly resistant to oxidants compared to non-malignant mesothelial cells. Since the level of cellular free radicals may be important for cell survival, we hypothesized that the increase of MnSOD in the mitochondria of mesothelioma cells may alter the free radical levels of these organelles. First, MnSOD activity was compared to the activities of two constitutive mitochondrial enzymes; MnSOD activity was 20 times higher in the mesothelioma cells than in the mesothelial cells, whereas the activities of citrate synthase and cytochrome c oxidase did not differ significantly in the two cell lines. This indicates that the activity of MnSOD per mitochondrion was increased in the mesothelioma cells. Superoxide production was assayed in the isolated mitochondria of these cells using lucigenin chemiluminescence. Mitochondrial superoxide levels were significantly lower (72%) in the mesothelioma cells compared to the mesothelial cells. Oxidant production in intact cells, assayed by fluorimetry using 2',7' dichlorodihydrofluorescein as a fluorescent probe, did not differ significantly between these cells. We conclude that mitochondrial superoxide levels are lower in mesothelioma cells compared to nonmalignant mesothelial cells, and that this difference may be explained by higher MnSOD activity in the mitochondria of these cells. Oxidant production was not different in these cells, which may be due to the previously observed increase in H2O2-scavenging mechanisms of mesothelioma cells. PMID- 10389974 TI - Glutathione-linked enzymes in benign and malignant oesophageal tissue. AB - Oxyradicals are involved in multiple mutational events and can contribute to the conversion of healthy cells to cancer cells. Glutathione (GSH) and the GSH replenishing enzymes keep the antioxidant status of normal cells at a level where they can avert oxyradical derived mutations. The aim of this study was to determine whether in cancer cells the GSH-replenishing, GSH antioxidant and GSH depleting enzymes were not at appropriate levels and therefore not able to protect cancer cells adequately against oxyradical-induced mutations. Cancer of the oesophagus was chosen since it is the most common gastrointestinal malignancy in South African Blacks. Biopsies and blood from 31 patients with cancer of the oesophagus and 29 non-cancer patients were assessed for these enzymes. The mean activity of the antioxidant and depleting enzyme GSH-peroxidase was elevated significantly by twofold in the cancer tissue compared to normal tissue. However, the activity of the replenishing enzyme GSSG-reductase and the level of the depleting enzyme GSH-s-transferase P1-isoenzyme were significantly reduced by 23% and 33% respectively. As in a previous paper we found that GSH was depleted and gamma-glutamine transpeptidase was diminished in oesophageal cancer. There can be two reasons for GSH depletion. Firstly, elevated GSH-peroxidase will use more GSH in an attempt to cope with the excessive production of oxyradicals as revealed by elevated lipid peroxidation; this was, as shown by us before, elevated sixfold in oesophageal cancer. Secondly, if little replenishment of GSH occurred the level of GSH would become lower. This was confirmed by our findings that the activities of the replenishing enzymes were significantly diminished in oesophageal cancer tissue. Contrary to what was expected, the other depleting enzyme GSH-s transferase P1 was not elevated in cancer tissue but was significantly lower. However, in the blood of the same patients it was significantly elevated. An explanation for this phenomenon is that, although the production of GST-P1 was enhanced in cancer, it did not show because it was rapidly extruded into the blood by an unknown mechanism operational only in cancer cells. PMID- 10389975 TI - In squamous cell carcinoma of the vulva, overexpression of p53 is a late event and neither p53 nor mdm2 expression is a useful marker to predict lymph node metastases. AB - To offer more tailored treatment to individual patients with squamous cell carcinoma of the vulva, more accurate prediction of lymph node metastases is required. As p53 and mdm2 are genes known to be involved in the development of other tumours, we studied expression of p53 and mdm2 in carcinogenesis of squamous cell carcinoma of the vulva and their clinical relevance. Archival material of 141 T1 and T2 vulvar tumours were used. Of the 141 primary tumours, the corresponding 39 lymph node metastases (LNM) were studied, and in 90 cases the pre-existent epithelia adjacent to the tumour (EAT) and in 14 cases vulvar intraepithelial neoplasia adjacent to the tumour (VIN) was also investigated. Detection of p53 and mdm2 protein was immunohistochemically performed. Scoring categories were: negative (1); weakly positive (2); moderately to markedly positive (3); and markedly positive (4). Overexpression of p53 was seen in 56% of the LNM, 39% of the primary tumours, 21% of the VIN lesions and 0% in the group of EAT. No relation was found between overexpression of p53 in the primary tumour and LNM. Expression of mdm2 was seen in 14% of the primary tumours, of which four cases were marked positive. In the group of LNM no mdm2-positive staining was observed. In the group of EAT, 25% was mdm2-positive, of which six cases were marked positive. In the group of VIN, 36% showed moderate (score 3) mdm2 expression. No relation was found between expression of mdm2 and LNM. In squamous cell carcinoma, overexpression of p53 is a late event in carcinogenesis. Marked expression of mdm2 is rarely seen in vulvar carcinomas, indicating that aberrant p53 cannot induce mdm2 expression. LNM cannot be predicted by detection of these proteins. PMID- 10389976 TI - Sequence analysis and transcript expression of the MEN1 gene in sporadic pituitary tumours. AB - The majority of pituitary tumours are monoclonal in origin and arise sporadically or occasionally as part of multiple endocrine neoplasia type 1 (MEN1). Whilst a multi-step aetiology involving both oncogenes and tumour suppressor genes has been proposed for their development, the target(s) of these changes are less clearly defined. Both familial and sporadic pituitary tumours have been shown to harbour allelic deletion on 11q13, which is the location of the recently cloned MEN1 gene. We investigated 23 sporadic pituitary tumours previously shown to harbour allelic deletion on 11q13 with the marker PYGM centromeric and within 50 kb of the MEN1 locus. In addition, the use of intragenic polymorphisms in exon 9 and at D11S4946, and of telomeric loci at D11S4940 and D11S4936, revealed that five of 20 tumours had loss of heterozygosity (LOH) telomeric to the menin gene. However, the overall pattern of loss in informative cases was indicative of non contiguous deletion that brackets the menin gene. Sequence analysis of all MEN1 coding exons and flanking intronic sequence, in tumours and matched patient leucocyte DNA, did not reveal mutation(s) in any of the 23 tumours studied. A benign polymorphism in exon 9 was encountered at the expected frequency, and in seven patients heterozygous for the polymorphism the tumour showed retention of both copies of the menin gene. Reverse transcription polymerase chain reaction analysis of ten evaluable tumours and four normal pituitaries revealed the presence of the menin transcript. Whilst these findings suggest that gene silencing is unlikely to be mechanistic in sporadic pituitary tumorigenesis, they do not exclude changes in the level or stability of the transcript or translation to mature protein. Our study would support and extend very recent reports of a limited role for mutations in the MEN1 gene in sporadic pituitary tumours. Alternatively, these findings may point to an, as yet, unidentified tumour suppressor gene in this region. PMID- 10389977 TI - DCC expression is related to mucinous differentiation but not changes in expression of p21(WAF1/Cip1) and p27Kip1, apoptosis, cell proliferation and human papillomavirus infection in uterine cervical adenocarcinomas. AB - To clarify possible roles of DCC expression in tumour differentiation and cell kinetics, we immunohistochemically investigated 80 uterine cervical adenocarcinomas (C-ACs), including 31 mucinous (M) and 31 endometrioid (E) lesions, and 18 adenocarcinomas in situ (AIS), along with 39 normal cervical samples. The results were compared with findings for p21(WAF1/Cip1) and p27Kip1 expression, apoptosis, cell proliferation and human papillomavirus (HPV) infection. Nine C-AC cases were also examined using a combination of the reverse transcription-polymerase chain reaction and Southern blot hybridization, as well as Western blot assays. Significantly decreased DCC scores were observed in E-ACs but not M-ACs, as compared to normal cervical glandular epithelia and AIS. Average p21(WAF1/Cip1) and p27Kip1 scores were significantly higher in E-ACs than M-ACs, in line with high apoptotic, mitotic and Ki-67 labelling indices. A concordance of the results for DCC and p21(WAF1/Cip1) expression between mRNA- and protein-based assays was also noted. Change of DCC expression, however, was not related to any of the cell kinetic markers or clinicopathological features in ACs of either type. There was also no association with the HPV status, although infection was significantly linked with high values for cell kinetics. These results suggest that DCC expression in C-ACs is closely associated with mucinous differentiation. PMID- 10389979 TI - The human herpesvirus-type 8 is not involved in malignant melanoma. AB - Malignant melanomas were supposed to harbour the human herpesvirus-type 8 (HHV-8) genome, as melanoma cells were reported to express interleukin-6 and a homologue of interleukin-6 was found in the HHV-8 genome. We therefore investigated 33 primary malignant melanomas by polymerase chain reaction, but could not find this tumorigenic gamma-herpesvirus in any tumour. PMID- 10389978 TI - Genetic alterations in hepatocellular carcinomas: association between loss of chromosome 4q and p53 gene mutations. AB - The major risk factors for hepatocellular carcinomas (HCC) in high incidence areas include infection with hepatitis B and C viruses (HBV, HCV) and exposure to aflatoxin. Genetic alterations in 24 liver resection specimens from Shanghai and Qidong were studied. Hepatitis B virus was integrated in all patient samples, and a null phenotype for the GSTM1 enzyme was present in 63% of patients. Alteration of p53 was present in 95% (23/24) of cases: mutations of the p53 gene in 12 HCC, p53 overexpression in 13 and loss of heterozygosity (LOH) of chromosome 17p in 17. All seven HCCs with a p53 mutation from Qidong and three of five from Shanghai had the aflatoxin-associated point mutation with a G to T transversion at codon 249, position 3. No HCC had microsatellite instability. LOH of chromosome 4q, 1p, 16q and 13q was present in 50%, 46%, 42% and 38%, respectively, and 4q was preferentially lost in HCCs containing a p53 mutation: LOH of 4q was present in 75% (9/12) of HCC with, but only 25% (3/12) of HCC without, a p53 gene mutation (P = 0.01). These data indicate a possible interaction between p53 gene mutation and 4q loss in the pathogenesis of HCC. PMID- 10389980 TI - Allele loss and mutation screen at the Peutz-Jeghers (LKB1) locus (19p13.3) in sporadic ovarian tumours. AB - Germline mutations in the LKB1 (STK11) gene (chromosome sub-band 19p13.3) cause characteristic hamartomas and pigmentation to develop in patients with Peutz Jeghers syndrome. Peutz-Jeghers syndrome carries an overall risk of cancer that may be up to 20 times that of the general population and Peutz-Jeghers patients are at increased risk of benign and malignant ovarian tumours, particularly granulosa cell tumours. Loss of heterozygosity (allele loss, LOH) has been reported in about 50% of ovarian cancers on 19p13.3. LKB1 is therefore a candidate tumour suppressor gene for sporadic ovarian tumours. We found allele loss at the marker D19S886 (19p13.3) in 12 of 49 (24%) sporadic ovarian adenocarcinomas. Using SSCP analysis, we screened ten ovarian cancers with LOH, 35 other ovarian cancers and 12 granulosa cell tumours of the ovary for somatic mutations in LKB1. No variants were detected in any of the adenocarcinomas. Two mutations were detected in one of the granulosa cell tumours: a mis-sense mutation affecting the putative 'start' codon (ATG --> ACG, M1T); and a silent change in exon 7 (CTT --> CTA, leucine). Like BRCA1 and BRCA2, therefore, it appears that LKB1 mutations can cause ovarian tumours when present in the germline, but occur rarely in the soma. The allele loss on 19p13.3 in ovarian cancers almost certainly targets a different gene from LKB1. PMID- 10389981 TI - Different gene expression of MDM2, GAGE-1, -2 and FHIT in hepatocellular carcinoma and focal nodular hyperplasia. AB - Overexpression and/or mutations of oncogenes, tumour suppressor genes and tumour rejection genes have been observed in several human malignancies. Their analyses might be of diagnostic importance. Therefore, malignant hepatocytes derived from hepatocellular carcinoma (HCC) tissue as well as non-malignant hepatocytes derived from focal nodular hyperplasia (FNH) were studied. Samples containing normal human hepatocytes (HC) served as controls. Cellular material was obtained by fine-needle aspiration biopsy guided by ultrasound. Cells were analysed for expression and mutation of the oncogene MDM2, the genes GAGE-1, -2 coding for tumour-associated antigens and the candidate tumour suppressor gene FHIT. Different patterns of non-mutant FHIT transcripts including precise deletion of exons were found in 7/10 HCC, 2/10 FNH and 2/10 HC. However, expression of non mutant GAGE-1, -2 RNA was demonstrated exclusively in 6/10 HCC samples. Further genetic features specific of HCC were point mutations in a zinc-finger motif of MDM2 (3/10 HCC samples). Neither GAGE-1, -2 expression nor MDM2 mutations were observed in the FNH samples, or in normal hepatocytes. Our findings suggest that occurrence of variable FHIT transcripts is not restricted to hepatic malignant tumours. In contrast, MDM2 mutations and GAGE-1, -2 expression were associated with HCC specimens. Therefore, the RT-PCR assays for GAGE-1, -2 and MDM2 might be useful adjuncts in cytodiagnosis of liver neoplasms. PMID- 10389982 TI - Role of p16/MTS1, cyclin D1 and RB in primary oral cancer and oral cancer cell lines. AB - One of the most important components of G1 checkpoint is the retinoblastoma protein (pRB110). The activity of pRB is regulated by its phosphorylation, which is mediated by genes such as cyclin D1 and p16/MTS1. All three genes have been shown to be commonly altered in human malignancies. We have screened a panel of 26 oral squamous cell carcinomas (OSCC), nine premalignant and three normal oral tissue samples as well as eight established OSCC cell lines for mutations in the p16/MTS1 gene. The expression of p16/MTS1, cyclin D1 and pRB110 was also studied in the same panel. We have found p16/MTS1 gene alterations in 5/26 (19%) primary tumours and 6/8 (75%) cell lines. Two primary tumours and five OSCC cell lines had p16/MTS1 point mutations and another three primary and one OSCC cell line contained partial gene deletions. Six of seven p16/MTS1 point mutations resulted in termination codons and the remaining mutation caused a frameshift. Western blot analysis showed absence of p16/MTS1 expression in 18/26 (69%) OSCC, 7/9 (78%) premalignant lesions and 8/8 cell lines. One cell line, H314, contained a frameshift mutation possibly resulting in a truncated p16/MTS1 protein. pRB was detected in 14/25 (56%) of OSCC but only 11/14 (78%) of these contained all or some hypophosphorylated (active) pRB. In premalignant samples, 6/8 (75%) displayed pRB, and all three normal samples and eight cell lines analysed contained RB protein. p16/MTS1 protein was undetectable in 10/11 (91%) OSCCs with positive pRB. Overexpression of cyclin D1 was observed in 9/22 (41%) OSCC, 3/9 (33%) premalignant and 8/8 (100%) of OSCC cell lines. Our data suggest p16/MTS1 mutations and loss of expression to be very common in oral cancer cell lines and less frequent in primary OSCC tumours. A different pattern of p16/MTS1 mutations was observed in OSCC compared to other cancers with all the detected p16/MTS1 mutations resulting in premature termination codons or a frameshift. The RB protein was expressed in about half (44%) of OSCCs and its expression inversely correlated with p16/MTS1 expression. In conclusion, we show that abnormalities of the RB pathway are a common mechanism of oral carcinogenesis. PMID- 10389983 TI - Swainsonine protects both murine and human haematopoietic systems from chemotherapeutic toxicity. AB - The haematopoietic system is sensitive to cytotoxic damage and is often the site of dose-limiting toxicity. We previously reported that swainsonine, an inhibitor of protein glycosylation, reduced the bone marrow toxicity resulting from a single dose of anticancer drugs in otherwise healthy mice. However, more important questions are (1) can swainsonine protect tumour-bearing mice without interfering with the anti-tumour effects of the drugs, and (2) can swainsonine stimulate haematopoietic activity of human, as well as murine, bone marrow. We demonstrate here that swainsonine protects C57BL/6 mice bearing melanoma-derived tumours from cyclophosphamide-induced toxicity without interfering with the drug's ability to inhibit tumour growth. Similar results were obtained in vivo with 3'-azido-3'-deoxythymidine (AZT), a myelosuppressive agent often used in therapy for acquired immune deficiency syndrome. Swainsonine increased both total bone marrow cellularity and the number of circulating white blood cells in mice treated with doses of AZT that typically lead to severe myelosuppression. Swainsonine also increased the number of erythroid and myeloid colony forming cells (CFCs) in short-term cultures of murine bone marrow, restoring the number of progenitor cells to the control level in the presence of AZT doses that reduced CFCs by 80%. With respect to the sensitivity of human haematopoietic cells to swainsonine, we show that swainsonine protected human myeloid progenitor cells from AZT toxicity in vitro. These results suggest that swainsonine may be useful as an adjuvant in several types of human chemotherapy. PMID- 10389984 TI - Vesnarinone and glucocorticoids cooperatively induce G1 arrest and have an anti tumour effect on human non-small cell lung carcinoma cells grown in nude mice. AB - Vesnarinone, an oral cardiotonic, inhibited the growth of several human non-small cell lung carcinoma cell lines, and its anti-proliferative effects in vitro and in vivo were greatly enhanced by combination with glucocorticoids, but not other steroids. Simultaneous treatment with vesnarinone and dexamethasone is the most effective to evoke the synergistic effect in the growth inhibition of lung carcinoma EBC-1 cells. Dexamethasone and other glucocorticoids induced morphological changes in EBC-1 cells and these agents together with vesnarinone induced alkaline phosphatase activity, which is a typical marker of type II pneumocyte maturation. This treatment arrested the growth of the cells at the G1 phase, indicating that this treatment is cytostatic rather than cytotoxic. These results suggest that vesnarinone plus glucocorticoid might be useful in lung cancer therapy. PMID- 10389985 TI - Modulation of oxygen consumption rate and vascular endothelial growth factor mRNA expression in human malignant glioma cells by hypoxia. AB - Cellular responses to hypoxia include modulation of respiration rate and up regulation of genes which encode for angiogenesis factors. We tested whether human malignant glioma cells vary in their response to hypoxic stress over the range of oxygen concentrations which exist in tumours. In five cell lines tested, decreased oxygen availability resulted in decreased rates of oxygen utilization, however substantial differences in the magnitude of the response were observed. Northern blot analysis was used to study induction of vascular endothelial growth factor mRNA in response to hypoxia. In two cell lines, modest hypoxia increased vascular endothelial growth factor mRNA levels compared with those of aerobic controls. In two additional cell lines, vascular endothelial growth factor mRNA was constitutively expressed under aerobic conditions and was not further increased by hypoxia. These findings demonstrate that differences in the response to hypoxia exist among human malignant glioma cell lines and suggest that therapies designed to exploit tumour hypoxia may have varying effects in tumours with different hypoxic stress responses. PMID- 10389986 TI - Anti-tumour promoter activity in Malaysian ginger rhizobia used in traditional medicine. AB - Zingiberaceae rhizomes commonly used in the Malaysian traditional medicine were screened for anti-tumour promoter activity using the short-term assay of inhibition of 12-O-tetradecanoyl phorbol-13-acetate (TPA)-induced Epstein-Barr virus early antigen (EBV-EA) in Raji cells. The inhibition of TPA-induced EBV-EA was detected using the indirect immunofluorescence assay (IFA) and Western blot technique. The indirect IFA detected the expression/inhibition of EBV-EA-D (diffused EA antigen), whereas the Western blot technique detected the expression/inhibition of both EBV-EA-D and EA-R (restricted EA antigen). Seven rhizomes were found to possess inhibitory activity towards EBV activation, induced by TPA; they are: Curcuma domestica, C. xanthorrhiza, Kaempferia galanga, Zingiber cassumunar, Z. officinale, Z. officinale (red variety), and Z. zerumbet. A cytotoxicity assay was carried out to determine the toxicity of the Zingiberaceae rhizome extracts. The rhizome extracts that exhibited EBV activation inhibitory activity had no cytotoxicity effect in Raji cells. Therefore, the present study shows that several Zingiberaceae species used in Malaysian traditional medicine contain naturally occurring non-toxic compounds that inhibit the EBV activation, which, if further investigated, could contribute in the development of cancer prevention methods at the tumour-promoting stage. PMID- 10389987 TI - The effects of hyperoxic and hypercarbic gases on tumour blood flow. AB - Carbogen (95% O2 and 5% CO2) has been used in preference to 100% oxygen (O2) as a radiosensitizer, because it is believed that CO2 blocks O2-induced vasoconstriction. However, recent work suggests that both normal and tumour arterioles of dorsal flap window chambers exhibit the opposite: no vasoconstriction vs constriction for O2 vs carbogen breathing respectively. We hypothesized that CO2 content might cause vasoconstriction and investigated the effects of three O2-CO2 breathing mixtures on tumour arteriolar diameter (TAD) and blood flow (TBF). Fischer 344 rats with R3230Ac tumours transplanted into window chambers breathed either 1%, 5%, or 10% CO2 + O2. Intravital microscopy and laser Doppler flowmetry were used to measure TAD and TBF respectively. Animals breathing 1% CO2 had increased mean arterial pressure (MAP), no change in heart rate (HR), transient reduction in TAD and no change in TBF. Rats breathing 5% CO2 (carbogen) had transiently increased MAP, decreased HR, reduced TAD and a sustained 25% TBF decrease. Animals exposed to 10% CO2 experienced a transient decrease in MAP, no HR change, reduced TAD and a 30-40% transient TBF decrease. The effects on MAP, HR, TAD and TBF were not CO2 dose-dependent, suggesting that complex physiologic mechanisms are involved. Nevertheless, when > or = 5% CO2 was breathed, there was clear vasoconstriction and TBF reduction in this model. This suggests that the effects of hypercarbic gases on TBF are site-dependent and that use of carbogen as a radiosensitizer may be counterproductive in certain situations. PMID- 10389988 TI - Suppression of metastasis-associated S100A4 gene expression by gamma-interferon in human colon adenocarcinoma cells. AB - S100A4 belongs to the S100 subfamily of calcium-binding proteins and has been suggested to be directly involved in invasion and metastasis of rodent and human tumour cells. The present study demonstrates that interferon gamma (IFN-gamma), but not IFN-alpha and IFN-beta, down-regulates the S100A4 mRNA level in colon adenocarcinoma WiDr cells in time- and dose-dependent manners. The effect was not associated with any cytotoxicity and was specific for the S100A4 mRNA, since the levels of the S100A6 and GAPDH mRNAs were not significantly affected by the treatment. IFN-gamma also strongly suppressed the S100A4 mRNA expression in HT-29 cells, but weakly in Colo201 cells. Flow cytometric analysis revealed that the level of the IFN-gamma receptor expression in Colo201 cells was lower than that in WiDr and HT-29 cells, suggesting that the suppression of the S100A4 expression by IFN-gamma depends on the amount of cell surface IFN-gamma receptor protein. IFN-gamma had no effect on the transcription rate of the S100A4 gene but reduced the stability of the S100A4 mRNA. WiDr cells treated with IFN-gamma showed reduced motile ability, further supporting the assumption that the S100A4 gene product is involved in controlling cell motility. PMID- 10389989 TI - Evaluation of the effects of photodynamic therapy with phosphorus 31 magnetic resonance spectroscopy. AB - Magnetic resonance spectroscopy in situ was used to study changes in phosphorus 31 metabolism after photodynamic therapy (PDT) of transplanted HeLa cell tumours. Tumours were irradiated 2 h after administration of ATX-S10 (8 formyloximethylidene-7-hydroxy-3-ethenyl-2,7,12,18, tetramethyl-porphyrin-13,17 bispropionil aspartate), a new photosensitizer and chlorin derivative. Nuclear magnetic resonance spectra were measured prior to illumination and 1, 3, 7, 14, 21 and 28 days after PDT on each mouse. A drastic decrease in adenosine triphosphate (ATP) and a concomitant increase in inorganic phosphate (Pi) were evident on the first day after PDT in all cases. The beta-ATP/total phosphate (P) ratio was 0.64 +/- 0.29% (average +/- s.d.) in complete response, 0.67 +/- 0.30% in recurrence and 2.45 +/- 0.93% in partial response. Comparison of this ratio to the histological findings revealed that the beta-ATP/total P ratio reflects the HeLa cell tumours which survived PDT. In other words, partial response on the one hand was distinguished from complete response and recurrence on the other with this ratio 1 day after PDT (P < 0.05). In addition, the ratio of phosphomonoester (PME) to Pi rose beyond 1.0 when macroscopic recurrence occurred, while it stayed under 1.0 in complete response. This finding suggests that the recurrence of HeLa cell tumours can be detected by the PME/Pi ratio. PMID- 10389990 TI - Effects of radiotherapy and estramustine on the microvasculature in malignant glioma. AB - Tumour angiogenesis is essential for progression of solid tumours and constitutes an interesting target for therapy. However, impaired tumour blood supply may also be an important obstacle for treatment by radiotherapy and chemotherapy. Estramustine has been shown to increase tumour blood flow and potentiate the effect of radiotherapy in experimental glioma. This study investigated the effects of fractionated radiotherapy and estramustine on angiogenesis in malignant glioma. The intracerebral BT4C rat glioma model was used and the animals were given whole brain radiotherapy 4 Gy x 5 days alone or in combination with estramustine 20 mg kg(-1) i.p. daily. Tumour microvascular density (MVD) was assessed by manual and computerized morphometrical analysis. Expression of vascular endothelial growth factor (VEGF) was studied by in situ hybridization. Radiotherapy decreased MVD to 157 vessels per mm2 compared to 217 vessels per mm2 in controls. Estramustine counteracted this anti-angiogenic effect and potentiated the anti-tumoural effect of radiotherapy. In addition, vessel size increased after estramustine treatment. Five days after completion of radiotherapy the expression of VEGF was increased in the centre of the tumours. In conclusion, fractionated radiotherapy decreases microvascular density in experimental malignant glioma. This effect was abolished by estramustine. The anti-vascular effect of irradiation is important to recognize when combining radiotherapy with cytotoxic drugs. PMID- 10389991 TI - Characterization of the model for experimental testicular teratoma in 129/SvJ mice. AB - An animal model of experimental testicular teratoma has been established to study how a teratoma affects the host testis and how the host testis reacts against the teratoma. 129/SvJ-mice were used as experimental animals. To induce the experimental testicular teratoma, male gonadal ridges from 12-day-old 129/SvJ mouse fetuses were grafted into the testes of adult mice for 1-12 weeks. The developing tumour was analysed by light and electron microscopy and by immunocytochemical localization of transcription factors SOX9 and c-kit, glial fibrillary acidic protein (GFAP) and type IV collagen. Testicular teratoma was observed in 36 out of 124 testes with implanted fetal gonadal ridges (frequency 29%). One spontaneous testicular teratoma was observed in this material from 70 male mice (1.5%). One week after implantation intracordal clusters of cells were seen in embryonic testicular cords of the graft as the first sign of testicular teratomas. Four weeks after implantation the embryonic testicular cords had totally disappeared from grafts with teratomas, and the tumour tissue had enlarged the testis and invaded the interstitium of the host testis. It consisted of solitary pieces of immature cartilage as well as of glial cells and of primitive neuroepithelium. Six to eight weeks after implantation the tumour tissue had expanded so that the enlarged testis could be detected by macroscopic enlargement of the scrotum. The testicular tissue of the host had practically disappeared, and only solitary disrupted seminiferous tubules of the host were seen surrounding the teratoma. Neuroepithelial structures of some teratomas cultured for 8 weeks had cells with a granular nucleus as a sign of obvious apoptosis. Eleven to 12 weeks after implantation the growth of the teratoma had stopped, and the histology corresponded to that of a mature cystic teratoma. GFAP, SOX9 and type IV collagen were strongly positive in some parts of the tumours cultured for 4 and 8 weeks, while only occasional c-kit-positive areas were observed in tumours cultured for 8 weeks. As conclusions: (1) the metastasizing capacity of the experimental testicular teratoma is very low during 12 weeks, but the behaviour of the tumour in the testicular tissue of the graft is invasive; (2) the growth of experimental testicular teratomas cease 6-8 weeks after implantation of the fetal gonadal ridges with the obvious apoptosis of the immature tissue components; (3) the model of experimental testicular teratoma in the mouse is suitable for studying how the teratoma affects the host testis and how the host testis reacts to teratoma. PMID- 10389992 TI - Prerequisites for effective isolated limb perfusion using tumour necrosis factor alpha and melphalan in rats. AB - An isolated limb perfusion (ILP) model using soft tissue sarcoma-bearing rats was used to study prerequisites for an effective ILP, such as oxygenation of the perfusate, temperature of the limb, duration of the perfusion and concentration of tumour necrosis factor (TNF). Combination of 50 microg TNF and 40 microg melphalan demonstrated synergistic activity leading to a partial and complete response rate of 71%. In comparison to oxygenated ILP, hypoxia was shown to enhance anti-tumour activity of melphalan alone and TNF alone but not of their combined use. Shorter perfusion times decreased anti-tumour responses. At a temperature of 24-26 degrees C, anti-tumour effects were lost, whereas temperatures of 38-39 degrees C or 42-43 degrees C resulted in higher response rates. However, at 42-43 degrees C, local toxicity impaired limb function dramatically. Synergy between TNF and melphalan was lost at a dose of TNF below 10 microg in 5 ml perfusate. We conclude that the combination of TNF and melphalan has strong synergistic anti-tumour effects in our model, just as in the clinical setting. Hypoxia enhanced activity of melphalan and TNF alone but not the efficacy of their combined use. For an optimal ILP, minimal perfusion time of 30 min and minimal temperature of 38 degrees C was mandatory. Moreover, the dose of TNF could be lowered to 10 microg per 5 ml perfusate, which might allow the use of TNF in less leakage-free or less inert perfusion settings. PMID- 10389993 TI - Cathepsin-D, urokinase plasminogen activator and type-1 plasminogen activator inhibitor in early breast cancer: an immunohistochemical study of prognostic value and relations to tenascin-C and other factors. AB - Cytosolic determinations of cathepsin-D (cath-D), urokinase plasminogen activator (uPA) and its specific inhibitor PAI-1 have shown an association with adverse prognosis in breast cancer. Our aim was to study the distribution of these markers in small axillary node-negative breast carcinomas using immunohistochemistry and relate the semiquantitative results to known prognostic factors, the expression of tenascin-C (Tn-C) in invasion border of the tumour and prognosis. All the 158 women (159 tumours) were treated with breast conserving surgery and postoperative radiotherapy. Cytoplasmic immunoreactivity for cath-D was seen in carcinoma cells in 47% and in stromal cells in 44%. Nearly all tumours expressed uPA and PAI-1, which were categorized to cytoplasmic expression in carcinoma cells and diffuse stromal expression and quantified -/+/++/ and further dichotomized for purposes of analysis. Expression of uPA and PAI-1 in stromal fibroblasts was recorded as -/+. Cytoplasmic and stromal cell cath-D contents were associated with grade, proliferation, Tn-C expression in the tumour invasion border and the development of distant metastasis. In multivariate analysis stromal cath-D proved to be an independent prognostic factor for metastasis. Stromal expression of uPA was associated with an increased risk of local recurrence; otherwise high levels of uPA did not associate with other prognostic factors nor with prognosis. Fibroblastic expression of PAI-1 showed an association with both local and distant disease recurrence. However, no consistent association between the immunohistochemically quantified uPA and PAI-1 and prognosis was found. In conclusion, immunohistochemical determination of cath D seems to be a viable method to predict a higher risk of metastasis but not local recurrence in small axillary node-negative breast carcinomas. PMID- 10389994 TI - Radioimmunotherapy of micrometastases in lung with vascular targeted 213Bi. AB - A model system has been used to test the efficacy of vascular targeting of alpha particle emitter 213Bi for therapy of small, 'artificial' metastases in mouse lung. Specific monoclonal antibody (mAb) 201 B was used to deliver greater than 30% of the injected dose to lung where tumours had developed due to intravenous injection of cells. Specific 213Bi-mAb 201B treatment of BALB/c mammary carcinoma EMT-6 tumours in lung resulted in a dose-dependent destruction of tumours and an extended lifespan of treated animals relative to controls. Significant reduction of lung tumour burden was noted in animals treated with 0.93 MBq injected dose or as little as 14 Gy absorbed dose to the lung. Animals treated with higher doses (2.6-6.7 MBq) had nearly complete cure of lung tumours but eventually died of lung fibrosis induced by the treatment. Four other tumour cell types were studied: murine Line 1 lung carcinomas in syngeneic BALB/c mice, rat IC-12 tracheal carcinoma growing in severe combined immune deficient (SCID) mice, and two human tumours--epidermoid carcinoma A431 and lung carcinoma A549--growing in SCID mice. In all cases, the number of lung tumour colonies was reduced in animals treated with specific, labelled mAb relative to those in animals treated with control 213Bi MAb or EDTA complexed 213Bi. Tumours treated in immunodeficient SCID mice were partially destroyed or at least retarded in growth, but ultimately regrew and proved fatal, indicating that an intact immune function is necessary for complete cure. The data show that the short-lived alpha particle emitter 213Bi can be effectively targeted to lung blood vessels and that tumour cells growing in the lung are killed. The mechanism may involve direct killing of tumour cells from alpha-particle irradiation, killing through destruction of blood supply to the tumour, or a combination of the two. PMID- 10389995 TI - Photodetection of early human bladder cancer based on the fluorescence of 5 aminolaevulinic acid hexylester-induced protoporphyrin IX: a pilot study. AB - Exogenous administration of 5-aminolaevulinic acid (ALA) is becoming widely used to enhance the endogenous synthesis of protoporphyrin IX (PpIX) in photodynamic therapy (PDT) and fluorescence photodetection (PD). Recently, results have shown that the chemical modification of ALA into its more lipophilic esters circumvents limitations of ALA-induced PpIX like shallow penetration depth into deep tissue layers and inhomogeneous biodistribution and enhances the total PpIX formation. The present clinical pilot study assesses the feasibility and the advantages of a topical ALA ester-based fluorescence photodetection in the human bladder. In this preliminary study 5-aminolaevulinic acid hexylester (h-ALA) solutions, containing concentrations ranging from 4 to 16 mM, were applied intravesically to 25 patients. Effects of time and drug dose on the resulting PpIX fluorescence level were determined in vivo with an optical fibre-based spectrofluorometer. Neither local nor systemic side-effects were observed for the applied conditions. All conditions used yielded a preferential PpIX accumulation in the neoplastic tissue. Our clinical investigations indicate that with h-ALA a twofold increase of PpIX fluorescence intensity can be observed using 20-fold lower concentrations as compared to ALA. PMID- 10389997 TI - Expression of nuclear retinoid receptors in normal, premalignant and malignant gastric tissues determined by in situ hybridization. AB - Retinoids exhibit multiple functions through interaction with nuclear retinoid receptors and have growth-suppressive activity on gastric cancer cells. To better understand the roles of nuclear retinoid receptors during gastric carcinogenesis, we have used in situ hybridization to investigate expression of retinoic acid receptors (RARs) and retinoid x receptors (RXRs) in premalignant and malignant formalin-fixed paraffin-embedded gastric tissues. Histological sections of eight normal, 17 distal normal and nine gastric cancer tissues were hybridized with non radioactive RNA probes for subtypes of RAR and RXR. Expression of RAR alpha, RAR beta, RAR gamma, RXR alpha and RXR beta was found in most cell types in gastric mucosa tissues from normal individuals as well as in distal normal tissues from cancer patients. Expression of RAR alpha and RAR beta were found in three and seven cancer tissues, respectively, and levels of RXR alpha mRNA were significantly decreased in poorly differentiated cancer tissues. Among the five investigated nuclear retinoid receptors, only expression of RAR alpha mRNA was significantly decreased in intestinal metaplasia, dysplasia and cancer tissues when compared to adjacent normal tissues. In conclusion, normal gastric mucosa expressed both RARs and RXRs, which supports the physiological role of retinoic acid on normal gastric mucosa. The decrease in RAR alpha expression in premalignant and malignant gastric tissues suggests a significant role of RAR alpha during gastric carcinogenesis. PMID- 10389998 TI - The hospital costs of treating lung cancer in the United Kingdom. AB - A detailed patient-by-patient costing analysis, based on case records for 253 patients diagnosed in 1993, reveals that the mean 4-year diagnosis and management costs amounted to Pound Sterling 6150 and Pound Sterling 5668 for non-small cell and small cell lung cancer respectively. These costs are lower than those identified in Canadian studies, the difference being explained by the use of a simulated costing methodology in these studies, lower unit costs and less aggressive interventions. PMID- 10389996 TI - Down-regulation of TGF-beta receptors in human colorectal cancer: implications for cancer development. AB - Many colorectal cancer cells are resistant to the anti-proliferative effects of transforming growth factor-beta (TGF-beta). TGF-beta also acts as paracrine factor from cancer cells on their mesenchymal cells. The aim of this study was to examine the expression of TGF-beta and its receptors in human colorectal cancer tissue and determine any relationship with cancer growth. In situ hybridization and Northern blot hybridization detection of TGF-beta1, type I and type II receptor mRNA and immunohistochemical staining of TGF-beta1 were performed using 11 human colorectal adenomas, 22 colorectal cancers and ten normal colorectal mucosas as control. TGF-beta receptor mRNAs were expressed mainly by normal colorectal epithelial cells and adenoma. However, mRNAs for TGF-beta receptors were only faintly, if at all, expressed in eight of 22 human colorectal cancers. In addition, intense signals of TGF-beta1 mRNA and the protein were detected in all colorectal cancers. TGF-beta receptor mRNAs and TGF-beta1 protein were also distributed in fibroblasts and endothelial cells in the interstitium. Moreover, Smad 4 protein was translocated to nucleus in primarily cultured adenoma cells, but not in cancer cells after TGF-beta stimulation. The escape of human colon cancer from TGF-beta-mediated growth inhibition by down-regulation of TGF-beta receptors as well as the effects of TGF-beta on stroma formation and angiogenesis indicate a possible role for TGF-beta in the progression of colon cancer in an intact host. PMID- 10389999 TI - Seroconversion after influenza vaccination in patients with lung cancer. AB - There are no data in the literature about the efficacy of influenza vaccination in lung cancer patients. Paired sera were available from 59 patients who received Fluvirin (Evans Medical). Of 41 patients susceptible to one or more influenza strains, 78% responded fully to vaccination. This response rate is comparable to that obtained from healthy volunteers. PMID- 10390000 TI - Assessment of bone response to systemic therapy in an EORTC trial: preliminary experience with the use of collagen cross-link excretion. European Organization for Research and Treatment of Cancer. AB - This study was designed to evaluate new bone resorption and tumour markers as possible alternatives to serial plain radiographs for the assessment of response to treatment. Thirty-seven patients with newly diagnosed bone metastases from breast cancer, randomized to receive oral pamidronate or placebo tablets in addition to anticancer treatment within the context of a multicentre EORTC trial, who were both assessable for radiographic response in bone and had serum and urine samples collected for more than 1 month were studied. The markers of bone metabolism measured included urinary calcium (uCa), hydroxyproline (hyp), the N telopeptide cross-links of type I collagen (NTx) and total alkaline phosphatase. The tumour markers measured were CA15-3 and cancer-associated serum antigen (CASA). Before treatment, levels of Ntx, uCa and Hyp were elevated in 41%, 24% and 28% respectively, and CA15-3 and CASA increased in 69% and 50%. For assessment of response and identification of progression, Ntx was the most useful bone marker. All markers behaved similarly in no change (NC) and partial response (PR) patients. There was a significant difference (P < or = 0.05) in Ntx levels (compared to baseline) at 1 and 4 months and in CA15-3/CASA at 4 months between patients with PR or NC and those with progressive disease (PD), and at 4 months between those with time to progression (TP) > 7 and those with TP < or = 7 months. The diagnostic efficiency (DE) for prediction of PD following a > 50% increase in Ntx or CA15-3 was 78% and 62% respectively. An algorithm to predict response to therapy has been developed for future prospective evaluation. PMID- 10390001 TI - Granulocyte colony-stimulating factor (G-CSF) transiently suppresses mitogen stimulated T-cell proliferative response. AB - Granulocyte colony-stimulation factor (G-CSF) is a cytokine that selectively promotes growth and maturation of neutrophils and may modulate the cytokine response to inflammatory stimuli. The purpose of this study was to examine the effect of G-CSF on ex vivo peripheral blood mononuclear cell (PBMC) functions. Ten patients with breast cancer were included in a clinical trial in which r metHuG-CSF was administered daily for 5 days to mobilize peripheral blood stem cells. Ten healthy women were also included as controls. Our data show that G-CSF treatment induces an increase in peripheral blood leucocyte, neutrophil, lymphocyte and monocyte counts. We have found a modulation in the percentages of CD19+, CD45+ CD14+, CD4+ CD45RA+ and CD4+ CD45RO+ cells in PBMC fractions during G-CSF treatment. We have also found a significant reduction in the proliferative response of PBMC to mitogenic stimulation that reverted 14 days after the fifth and the last dose of G-CSF. Furthermore, it was not associated with significant changes in the pattern of cytokine production. The mechanism of this immunoregulatory effect is probably indirect since G-CSF receptor has not been found in T lymphocytes. This mechanism and its potential clinical applications remain to be elucidated. PMID- 10390002 TI - Effects of radiotherapy after hyperbaric oxygenation on malignant gliomas. AB - The purpose of this non-randomized trial was to evaluate the efficacy of radiotherapy combined with hyperbaric oxygen (HBO) in patients with malignant glioma. Between 1987 and 1997, 29 patients in whom computerized tomography (CT) or magnetic resonance imaging (MRI) scans showed post-operative residual tumours were locally irradiated with nitrosourea-based chemotherapy. Treatments were consecutively combined with HBO at two institutions since 1991 and 1993. Fifteen patients were irradiated daily after HBO, and the periods of time from decompression to irradiation were within 15 and 30 min in 11 and four patients respectively. Fourteen other patients were treated without HBO. Tumour responses were assessed by CT or MRI scans and survival times were compared between the treated groups. In the HBO group, 11 of 15 patients (73%) showed > or = 50% tumour regression. All responders were irradiated within 15 min after decompression. In the non-HBO group, four of 14 patients (29%) showed tumour regression. The median survivals in patients with and without HBO were 24 and 12 months, respectively, and were significantly different (P < 0.05). No serious side-effects were observed in the HBO patients. In conclusion, irradiation after HBO seems to be a useful form of treatment for malignant gliomas, but irradiation should be administered immediately after decompression. PMID- 10390003 TI - Promoting patient participation in the cancer consultation: evaluation of a prompt sheet and coaching in question-asking. AB - Active participation in the medical consultation has been demonstrated to benefit aspects of patients' subsequent psychological well-being. We investigated two interventions promoting patient question-asking behaviour. The first was a question prompt sheet provided before the consultation, which was endorsed and worked through by the clinician. The second was a face to face coaching session exploring the benefits of, and barriers to, question-asking, followed by coaching in question-asking behaviour employing rehearsal techniques. Sixty patients with heterogeneous cancers, seeing two medical oncologists for the first time, were randomly assigned to one of three groups: two intervention groups and one control group. Sociodemographic variables and anxiety were assessed prior to the intervention which preceded the consultation. The consultations were audiotaped and subsequently analysed for question-asking behaviour. Anxiety was assessed again immediately following the consultation. Questionnaires to assess patient satisfaction, anxiety and psychological adjustment were sent by mail 2 weeks following the consultation. Presentation and discussion of the prompt sheet significantly increased the total number of questions asked and the number of questions asked regarding tests and treatment. Coaching did not add significantly to the effects of the prompt sheet. Psychological outcomes were not different among the groups. We conclude that a question prompt sheet addressed by the doctor is a simple, inexpensive and effective means of promoting patient question asking in the cancer consultation. PMID- 10390004 TI - Ejaculation in testicular cancer patients after post-chemotherapy retroperitoneal lymph node dissection. AB - The purpose of this study was to evaluate fertility after different types of post chemotherapy retroperitoneal lymph node dissection (RPLND). During 1980-1994, 192 patients with metastatic testicular cancer underwent post-chemotherapy RPLND with a gradual shift from modified bilateral template RPLND to nerve-sparing RPLND. Modified bilateral template RPLND was done in 92% of the patients operated during 1980-1984 as compared to 16% during 1989-1994. Pre- and post-treatment fertility was assessed by microscopic sperm analysis, determination of serum FSH and information on ejaculation and paternity. There was no significant difference of the survival rates between the three treatment periods. Antegrade ejaculation was preserved in 11% of the patients after modified bilateral template RPLND as compared to 89% after the nerve-sparing operation technique. The median ejaculatory volume decreased post-operatively, serum FSH increased and sperm density remained unchanged. Fifty-six patients attempted fatherhood after their treatment, and 27 fathered at least one child after an observation-time of 55 months, nine of them by assisted fertilization. Patients with initially advanced testicular cancer but limited residual retroperitoneal masses after induction chemotherapy can safely undergo limited post-chemotherapy RPLND as a part of multimodality treatment. After nerve-sparing RPLND antegrade ejaculation is preserved in 89% of the patients though the ejaculatory volume decreases after RPLND. Post-treatment fatherhood can be achieved in at least 50% of the patients attempting paternity. PMID- 10390005 TI - Comparison of deregulated expression of cyclin D1 and cyclin E with that of cyclin-dependent kinase 4 (CDK4) and CDK2 in human oesophageal squamous cell carcinoma. AB - The expressions of cyclin D1, cyclin E, cyclin-dependent kinase 4 (CDK4), and CDK2 were immunohistochemically examined in 90 patients with human oesophageal squamous cell carcinoma (SCC) to determine their relationship to the tumour behaviour and patient prognosis. Nuclear immunostaining of cyclin D1 and cyclin E was observed in 28 (31.1%) and 27 tumours (30.0%) respectively. Thirty-nine tumours (43.3%) and 31 tumours (34.4%) exhibited both cytoplasmic and nuclear positivity for CDK4 and CDK2 respectively. Of 28 cyclin D1-positive and 27 cyclin E-positive tumours, CDK4 was overexpressed in 12 (42.8%) tumours and CDK2 in seven (25.9%) tumours respectively. There was no significant relationship in immunopositivity between cyclin D1 and CDK4 or between cyclin E and CDK2. Simultaneous immunoreactivity for both cyclin D1 and CDK4 was significantly associated with venous invasion (P < 0.05). In a univariate analysis, the prognosis of patients with tumours that were both cyclin D1- and CDK4-positive was significantly poorer than that of patients with cyclin D1-negative tumours (P < 0.05). In a multivariate analysis, both cyclin D1 and CDK4 immunoreactivities (P < 0.01) and tumour stage (P < 0.001) were recognized as independent risk factors. In this analysis, the hazard ratio for cyclin D1-positive and CDK4 negative cases compared with cyclin D1-negative cases was significant (hazard ratio = 3.128, 95% confidence interval = 1.418-6.899, P = 0.0047). No significant prognostic relevance was detected in both cyclin E and CDK2 immunoreactivity. Our in vivo findings suggest that in human oesophageal SCC, cyclin D1 and cyclin E and their functional partners, CDK4 and CDK2, often exhibit dysregulated overexpression in many cases, and that tumours with simultaneous expression of cyclin D1 and CDK4 are frequently associated with venous invasion and have a worse prognosis, statistically. Moreover, overexpression of cyclin D1 alone may also contribute to tumour progression independent of CDK4 overexpression. PMID- 10390006 TI - Psychological, clinical and pathological effects of relaxation training and guided imagery during primary chemotherapy. AB - The diagnosis and treatment of breast cancer are stressful, and stress may be associated with a poorer response to chemotherapy. There is a need, therefore, to develop and evaluate interventions that might enhance quality of life and, possibly, improve treatment response. The effects of relaxation combined with guided imagery (visualizing host defences destroying tumour cells) on quality of life and response to primary chemotherapy, to date, have not been adequately evaluated. Ninety-six women with newly diagnosed large or locally advanced breast cancer (T2 > 4 cm, T3, T4, or TxN2 and M0) took part in a prospective, randomized controlled trial. Patients were randomized following diagnosis to a control condition (standard care) or to the experimental condition (standard care plus relaxation training and imagery). Psychometric tests to evaluate mood and quality of life were carried out before each of the six cycles of chemotherapy and 3 weeks after cycle 6: tests of personality and coping strategy were carried out prior to cycles one and six. Clinical response to chemotherapy was evaluated after six cycles of chemotherapy using standard UICC criteria and pathological response was assessed from the tissue removed at surgery. As hypothesized, patients in the experimental group were more relaxed and easy going during the study (Mood Rating Scale). Quality of life was better in the experimental group (Global Self-assessment and Rotterdam Symptom Checklist). The intervention also reduced emotional suppression (Courtauld Emotional Control Scale). The incidence of clinically significant mood disturbance was very low and the incidence in the two groups was similar. Finally, although the groups did not differ for clinical or pathological response to chemotherapy, imagery ratings were correlated with clinical response. These simple, inexpensive and beneficial interventions should be offered to patients wishing to improve quality of life during primary chemotherapy. PMID- 10390007 TI - Long-term survival after epirubicin, cisplatin and fluorouracil for gastric cancer: results of a randomized trial. AB - We report the final results of a prospectively randomized study that compared the combination of epirubicin, cisplatin and protracted venous infusion fluorouracil (5-FU) (ECF regimen) with the standard combination of 5-FU, doxorubicin and methotrexate (FAMTX) in previously untreated patients with advanced oesophagogastric cancer. Between 1992 and 1995, 274 patients with adenocarcinoma or undifferentiated carcinoma were randomized from eight oncology centres in the UK and analysed for response and survival. The overall response rate was 46% (95% confidence interval (CI), 37-55%) with ECF, and 21% (95% CI, 13-28%) with FAMTX (P = 0.00003). The median survival was 8.7 months with ECF and 6.1 months with FAMTX (P = 0.0005). The 2-year survival rates were 14% (95% CI, 8-20%) for the ECF arm, and 5% (95% CI, 2-10%) for the FAMTX arm (P = 0.03). Histologically complete surgical resection following chemotherapy was achieved in ten patients in the ECF arm (three pathological complete responses to chemotherapy) and three patients in the FAMTX arm (no pathological complete responses). The ECF regimen resulted in a response and survival advantage compared with FAMTX chemotherapy. The probability of long-term survival following surgical resection of residual disease is increased by this treatment. The high response rates seen with ECF support its use in the neoadjuvant setting. PMID- 10390008 TI - Urinary concentrations of the soluble adhesion molecule E-cadherin and total protein in patients with bladder cancer. AB - Reduced expression of the adhesion molecule E-cadherin has been associated with increased invasiveness and poorer survival in patients with bladder cancer. We have examined soluble E-cadherin (sE-cadherin) and total protein concentrations in urine from patients with bladder cancer (n = 34), non-neoplastic benign urological diseases (n = 14) and healthy controls (n = 21) to determine their diagnostic and prognostic significance. Soluble E-cadherin concentrations of the cancer group were significantly higher (P < 0.001) than those of the controls but the benign group was not significantly different from either the cancer group or the controls. When sE-cadherin concentrations were adjusted for creatinine, similar but more statistically significant results were obtained and the benign group was significantly elevated compared with the controls (P < 0.01). No differences were apparent between the invasive (pT1-4) and non-invasive (pTa) cancers. Urinary total protein concentrations in the cancer group were significantly higher than the controls (P < 0.001) and the benign group (P < 0.05) although no difference was seen between the benign group and patients with non-invasive (pTa) cancer or between the benign group and controls. When expressed as the protein/creatinine index, results were similar but more statistically significant and a significant difference was seen between invasive and non-invasive cancers (P < 0.01). Only the protein/creatinine index correlated significantly with stage of the tumour (P < 0.01). It is concluded that urinary sE-cadherin measurements are of no greater value than urinary total protein. PMID- 10390009 TI - The significance of measuring monocyte tissue factor activity in patients with breast and colorectal cancer. AB - Monocytes express tissue factor (mTF) in several conditions including cancer where levels may be valuable in assessing tumour presence and progression. Using a two-stage kinetic chromogenic assay (KCA), mTF levels were measured in controls [normal subjects (n = 60) and patients undergoing hernia repair or cholecystectomy (n = 60)], in patients with benign and malignant disease of the breast (n = 83) and of the large bowel (n = 62). This was performed under fresh (resting) conditions and after incubation for 6 h without (unstimulated) and with (stimulated) Escherichia coli endotoxin. The malignant groups showed higher mTF levels than each of the three controls for resting (P < 0.05 breast, P < 0.05 colorectal) unstimulated (P < 0.05 breast, P < 0.05 colorectal) and stimulated cells (P < 0.001 breast, P < 0.01 colorectal). Similarly, the benign inflammatory groups had higher mTF levels than controls for resting (P < 0.05 colorectal), unstimulated (P < 0.05 colorectal) and stimulated cells (P < 0.01 breast, P < 0.01 colorectal). There was no significant difference between malignant and benign inflammatory groups in each organ. mTF levels showed an increase corresponding to that of histological tumour progression and were higher in non surviving patients. In conclusion, mTF levels are raised in malignant and inflammatory disease compared to controls and patients with non-inflammatory conditions. Stimulated cells give better discrimination between the groups and may be of value in identifying high risk individuals. mTF levels showed an association with tumour grade or stage and the patients' survival time. PMID- 10390010 TI - Prognostic value of tissue-type plasminogen activator (tPA) and its complex with the type-1 inhibitor (PAI-1) in breast cancer. AB - The prognostic value of tissue-type plasminogen activator (tPA) measured in samples derived from 865 patients with primary breast cancer using a recently developed enzyme-linked immunosorbent assay (ELISA) was evaluated. Since the assay could easily be adapted to the assessment of the complex of tPA with its type-1 inhibitor (PAI-1), it was investigated whether the tPA:PAI-1 complex also provides prognostic information. To this end, cytosolic extracts and corresponding detergent extracts of 100,000 g pellets obtained after ultracentrifugation when preparing the cytosolic fractions for routine steroid hormone receptor determination were assayed. Statistically significant correlations were found between the cytosolic levels and those determined in the pellet extracts (Spearman correlation coefficient r(s) = 0.75, P < 0.001 for tPA and r = 0.50, P < 0.001 for tPA:PAI-1 complex). In both Cox univariate and multivariate analysis elevated levels of (total) tPA determined in the pellet extracts, but not in cytosols, were associated with prolonged relapse-free (RFS) and overall survival (OS). In contrast, high levels of the tPA:PAI-1 complex measured in cytosols, but not in the pellet extracts, were associated with a poor RFS and OS. The prognostic information provided by the cytosolic tPA:PAI-1 complex was comparable to that provided by cytosolic (total) PAI-1. Furthermore, the estimated levels of free, uncomplexed tPA and PAI-1, in cytosols and in pellet extracts, were related to patient prognosis in a similar way as the (total) levels of tPA and PAI-1 respectively. Determination of specific forms of components of the plasminogen activation system, i.e. tPA:PAI-1 complex and free, uncomplexed tPA and/or PAI-1, may be considered a useful adjunct to the analyses of the separate components (tPA and/or PAI-1) and provide valuable additional prognostic information with respect to survival of breast cancer patients. PMID- 10390011 TI - Mutation testing in melanoma families: INK4A, CDK4 and INK4D. AB - The INK4A gene which codes for the cyclin-dependent kinase (CDK) inhibitor INK4A or p16 underlies susceptibility to melanoma in some families. Germline mutations in the gene that codes for the target protein of p16, CDK4, underlie susceptibility in very rare families. We report mutation screening of the INK4A and CDK4 genes in 42 UK families. A total of nine families were identified with INK4A mutations and none with CDK4 exon 2 mutations. These mutations were in 8/22 (35%) families with three or more cases of melanoma and 1/20 (5%) families with only two cases. In one of these nine families a novel single base pair substitution was identified, Gly67Arg. In an attempt to identify another melanoma susceptibility gene, a member of the INK4 family, the p19 INK4D gene has been studied. The p19 gene was sequenced in DNA from the 42 UK families and six additional US families. No mutations were identified. PMID- 10390014 TI - A simple audio data logger for objective assessment of snoring in the home. AB - We have developed a portable device for patient use in logging snoring loudness in the home, for guiding treatment decisions and measuring the clinical effectiveness of treatment. The device uses a free field microphone and is positioned on a bedside table. The prototype devices contain no inherently expensive components and are simple to operate (producing only 5% patient error to date). They are portable, battery powered, rugged and produce digital data which are easily and automatically analysed, and these design parameters enable the devices to be used for first line patient assessment. Of the 75 recordings made so far from 30 patients, 85% were successful, yielding clinically useful data. Because it is sound levels which are recorded and not replayable sounds, patient privacy is maintained, resulting in excellent patient acceptance (to date no patient has refused). The device has a dynamic range of 45-90 dB sound pressure level and a frequency range of 30 Hz-5 kHz. Because snoring intensities often vary significantly throughout the night the device can measure continuously over 8 h. PMID- 10390013 TI - Vascular endothelial growth factor-C expression in human prostatic carcinoma and its relationship to lymph node metastasis. AB - Lymph node dissemination is a major prognostic factor in human cancer. However, the molecular mechanisms underlying lymph node metastasis are poorly understood. Recently, vascular endothelial growth factor-C (VEGF-C) was identified as a ligand for VEGF receptor-3 (VEGFR-3/Flt-4) and the expression of VEGFR-3 was found to be highly restricted to the lymphatic endothelial cells. In this report, we investigated the expression of VEGF-C and VEGFR-3 in human prostatic carcinoma tissue by using in situ hybridization and immunohistochemical staining respectively. Expression of VEGF-C mRNA in prostatic carcinoma was significantly higher in lymph node-positive group than in lymph node-negative group. In addition, the number of VEGFR-3-positive vessels was increased in stroma surrounding VEGF-C-positive prostatic carcinoma cells. These results suggest that the expression of VEGF-C in prostatic carcinoma cells is implicated in the lymph node metastasis. PMID- 10390012 TI - Expression of MUC1 mucins inversely correlated with post-surgical survival of renal cell carcinoma patients. AB - Surgical specimens of the normal kidney and of renal cell carcinoma (RCC) tissues at different stages of progression and of various histological grades were examined for the expression of MUC1 mucins with sialylated carbohydrates (sialylated MUC1 mucins) using a monoclonal antibody MY.1E12. Immunohistochemical studies revealed that the binding sites for this antibody were localized to the apical side of the epithelial cells of the distal convoluted tubules, Henle's loops and collecting ducts. However, proximal convoluted tubules, where RCC is considered to originate, were not stained. This antibody also bound strongly to RCC at advanced stages of progression and at metastatic sites, and to RCC of histologically high grades (undifferentiated). The epitope, presumably sialylated MUC1 mucin, was detected not only along the surface of the cell membranes but also in the cytoplasm. The level of expression of sialylated MUC1 mucins was inversely correlated with the survival of the patients with RCC and the disease free survival period after curative surgery. Western blot analysis demonstrated that the electrophoretic mobility of sialylated MUC1 mucins of RCC was greater than that from the normal kidney. It is suggested that high levels of expression of sialylated MUC1 mucins in certain human RCC populations correlate with the aggressiveness of the disease, such as the tendency to form metastasis. PMID- 10390015 TI - A new method for continuous tonometric pCO2 measurement--in vitro studies. AB - The available methods for tonometric pCO2 measurement only provide the possibility of performing intermittent registrations. A new method allowing continuous tonometric pCO2 measurement has been developed and tested in an in vitro model. A standard tonometer for intestinal pCO2 measurement was modified to allow continuous perfusion of the balloon with physiological saline solution in a closed system. The pCO2 in the system was determined in a specially constructed measurement chamber with a TCM20 percutaneous pCO2 monitor. In this in vitro model the tonometer balloon was placed in a saline bath with a constant pCO2 concentration and the measurements from the closed circulating system were compared with those obtained from a standard tonometer placed in the same bath. In 8 and 24 h experiments the circulating system measured the pCO2 value as accurately and reliably as traditional tonometry. This study indicates that the new method makes continuous monitoring of pCO2 possible. PMID- 10390016 TI - Detection of body movements during sleep by monitoring of bed temperature. AB - We have modified a previously developed bed temperature monitor and used it to evaluate a sleeper's body movements. Sleep was monitored both at home and in the laboratory in normal subjects. From laboratory monitoring, a new algorithm to assess body movements using the sum of a square of temperature differences (SSD(t)) method was proposed, and the best conditions for detection of body movements are discussed. The experiment was performed with 12 normal male subjects and the body movement obtained from temperature changes was compared with that from video images. The sensitivity and positive predictive accuracy of body movement at the lower limbs with a sampling interval of 15 s and a threshold value of 0.2 degrees C2 were 88.7% and 95.4% respectively, over a total of 22 nights' observation. The results show that body movement can be accurately detected by placing one belt sensor under the lower limbs with a sampling interval of 15 s and a threshold value of 0.2 degrees C2 in SSD(t) analysis. We also found that body movement frequency is different among individuals during sleep, but has a certain range for an individual. No significant relationship between average body movement or average time in bed (TIB) and sleep profiles was observed among individuals for all subjects in one week of home monitoring. However, significant relationships between frequency of body movement or TIB and sleep profiles were observed in the long-term home monitoring for an individual. This suggests that body movements and TIB detected by bed temperature measurement can be used as indices for the assessment of sleep stability and continuity. PMID- 10390017 TI - Duodenogastric reflux of bile in health: the normal range. AB - Duodenogastric reflux (DGR) is suspected to be an aetiological factor in the pathogenesis of foregut disease. The 'Bilitec' bile probe allows continuous detection of bilirubin, based on spectrophotochemical properties. We aimed to describe duodenogastric bile reflux in healthy, normal volunteers in a Western European population, as a basis for the future study of DGR in disease. An international multicentre study was established. DGR was measured using 24 h ambulatory bile and pH monitoring in the proximal stomach, in 43 normal volunteers from the third to the seventh decades. Subjects adhered to a standard protocol. The total test period, supine and upright components, were analysed. The 90th percentile values for absorbance thresholds of 0.14, 0.25, 0.3, 0.4 and 0.5 were 40.5%, 20.9%, 19.6%, 11.6% and 4.6% of the total time respectively. There was a wide range of absorbance within each threshold. Supine DGR was greater than upright, and associated with an alkaline tide. The upright phase was further subdivided into upright fasting, prandial and post prandial phases, and ranges for these periods are also described. No relationship between age, weight, or body mass index and duodenogastric reflux was seen. The results of this study form a range which allows further investigation into the contribution of duodenogastric bile reflux in the pathogenesis of foregut disease. PMID- 10390018 TI - Coherence between body surface ECG leads and intracardiac signals increases during the first 10 s of ventricular fibrillation in the human heart. AB - Ventricular fibrillation (VF) in the human heart is not well understood. The aim of this study was to measure changes in the phase relationship between the body surface ECG and intracardiac electrograms recorded during the first 10 s of human VF. We studied 11 episodes of VF and measured the coherence of (a) ECG lead I and ECG lead V1, (b) ECG lead V1 and the right ventricular apex (RVA) electrogram, and (c) ECG lead V1 and the smoothed RVA electrogram. Each coherence measurement was the average of the magnitude squared coherence function in the range 0-60 Hz, and measurements were made 1, 3, 5, 7 and 9 s after the onset of VF. Overall, the mean (SD) coherence was 31(6)% between ECG leads I and V1, 17(3)% between ECG lead V1 and the RVA electrogram, and 20(4)% between ECG lead V1 and the smoothed RVA electrogram. All three measurements of coherence increased significantly between 1 and 9 s with mean (SD) rates of 0.97(1.01)% s(-1), 0.8(1.18)% s(-1) and 0.82(1.19)% s(-1) respectively. These results show that propagation in human VF becomes more organized during the first 10 s of VF. This may be an optimal window for defibrillation. PMID- 10390019 TI - Palatal snoring identified by acoustic crest factor analysis. AB - The differentiation of palatal from non-palatal snoring is very important for ENT surgeons trying to determine whether palatal surgery would be curative. At present sleep nasendoscopy is the accepted method. Palatal vibration produces marked modulation of sound loudness at low frequency (below approximately 100 Hz). We calculate a crest factor for the sound waveform (ratio of peak to root mean square (rms) value in any given epoch), as a measure of the degree of modulation. Free-field snore sounds were recorded from 11 supine adult patients under intravenous sedation (midazolam), using a digital tape recorder. Recordings were transferred to a PC (sampling frequency 11 kHz), and analysed using code written by us. Direct visual confirmation of the site of snoring was gained from simultaneous sleep nasendoscopy, taken as the gold standard. In six patients the dominant site was the soft palate. The non-palatal group (five patients) comprised one epiglottic, two hypopharyngeal and two tongue base snorers. The crest factor was found to be significantly higher for palatal snorers (p < 0.01, Student-t or Mann-Whitney tests). Furthermore, palatal could be distinguished from non-palatal snorers on the basis of crest factor alone in all 11 cases, making this a promising non-invasive diagnostic technique. PMID- 10390020 TI - Inductive plethysmography components analysis and improved non-invasive postoperative apnoea monitoring. AB - Twenty-nine patients were monitored overnight for breathing distress patterns during postoperative analgesia. Nasal flow apnoea monitoring and pulse oximetry data were recorded at 50 Hz. Respiratory inductive plethysmography (RIP) tracked tidal volume (TV) thoracoabdominal motion, and supplemented the flow monitoring by identifying detected apnoea type. TVs were computed from linear combinations of the RIP signals, but calibrations showed that multiple regression approaches with fitting errors <1% had highly variable coefficients and estimate precisions. Simple least squares theory showed that unstable parameter calculation and coefficient variation with signal conditions were inherent in RIP calibration models. Principal components (PC) methods were well suited to mitigating these problems because the RIP signals were highly correlated. The two PCs tracked the relative changes in TVs and indicated the degree of signal asynchrony, enabling improved uncalibrated monitoring. For accurately measuring RIP phase differences, the cross-correlation function was calculated. A simple version of PC analysis is developed, avoiding matrices, to help clarify how RIP calibration problems can be addressed. The methods are illustrated for calibration in normal breathing, and for postoperative monitoring during Cheyne-Stokes breathing. Sum and difference combinations of the RIP signals could discriminate central from obstructive apnoeas to help improve flow monitoring efficacy on-line. PMID- 10390021 TI - Separation of haemodynamic flow waves measured by MR into forward and backward propagating components. AB - Physiological information on the action of the heart and on the reflection sites in the arterial system can be derived respectively from the forward and the backward propagating pressure or flow wave components. Earlier work on the separation of these components was exclusively based on invasive measurements of pressure or flow. In this study magnetic resonance (MR), which is a non-invasive imaging technique, was used to measure the blood flow waveform simultaneously at multiple positions along a vessel. Linear one dimensional transmission-line theory was used to separate the flow waves into forward and backward propagating components. First results, obtained from the thoracic aorta of five healthy male volunteers, consistently showed a negative reflection with a delay of about 100 ms between the foot of the forward and the foot of the backward propagating flow wave. Our model, consisting of a single vessel segment with constant diameter and wall properties, was validated by the excellent agreement between the vessel area as calculated from the flow data using the law of mass conservation and as directly measured with a different independent MR technique. PMID- 10390022 TI - Prediction of prolonged ventilator dependence in children by respiratory function measurements. AB - Complications of ventilatory support are more common if this assistance is prolonged. Our aim was to determine if results of respiratory function measurement on the first day of ventilation identified children who would develop prolonged ventilatory dependence (> or = 4 days) and whether such results were a more accurate predictor than readily available clinical data. Thirty three children, median age 2 years (range 0.1-13.6), who were supported by a constant flow ventilator and hence had measurements of compliance of the respiratory system (CRS) and resistance of the respiratory system (RRS) on the first day of ventilatory support, were retrospectively identified. Those who needed prolonged ventilatory support had a lower CRS on day one (p < 0.01) and required at any time during their ventilatory career both a higher maximum inspired oxygen concentration (p < 0.01) and peak inspiratory pressure (PIP) (p < 0.01). Logistic regression analysis demonstrated that only a low CRS and high maximum PIP were significantly correlated with prolonged ventilator dependence. A low CRS (<0.4 (ml/cmH2O) kg(-1)) and a high maximum PIP (>27 cmH2O) had similar sensitivities (83%) and specificities (71% and 67% respectively) in predicting prolonged ventilatory dependence. The CRS results, unlike the maximum PIP results, however, were always available on the first day of ventilatory support. We therefore conclude that respiratory function measurements have a role in identifying children who would benefit from strategies to prevent prolonged ventilator dependence. PMID- 10390023 TI - Transit time heterogeneity of bolus flow through the heart and lungs in patients with left-to-right intracardiac shunt. AB - Knowledge of the contributions of transit time heterogeneity to the cardiopulmonary system is important for understanding cardiopulmonary function in patients with intracardiac shunt. We determined the heterogeneity of blood transit times occurring between the right atrium and the left ventricle. Eighty two patients with suspected left-to right shunt were investigated with first-pass 99mTc-labelled red blood cell radiocardiography at supine rest. Forty two of them had a pulmonary-to-systemic flow ratio (Qp:Qs) of less than 1.2 and they served as a control group. The remaining study subjects had a Qp:Qs ratio of 1.7 +/- 0.3 (mean +/- SD). The patients with shunt had significantly greater (p < 0.001) heterogeneity of transit times (49 +/- 9%) than in the controls (39 +/- 7%). Overall heterogeneity of cardiopulmonary transit times in patients with shunt showed a curvature relationship; the highest values were centred in patients with moderate to severe shunt (1.5 < Qp:Qs < 2.5). The results suggest that the increased heterogeneity of transit times mainly occurs within the pulmonary capillary bed in patients with intracardiac shunt. This is probably due to the recruitment of the open capillaries without vessel distension. PMID- 10390024 TI - Reorganized cerebral metabolic interactions in temporal lobe epilepsy. AB - Patients with left temporal lobe epilepsy demonstrate language impairments that are not well understood. To explore abnormal patterns of brain functional connections with respect to language processing, we applied a principal component analysis to resting regional cerebral metabolic data obtained with positron emission tomography in patients with right- and left-sided temporal lobe epilepsy and controls. Two principal components were expressed differentially among the groups. One principal component comprised a pattern of metabolic interactions involving left inferior frontal and left superior temporal regions-corresponding to Broca's and Wernicke's areas, respectively-and right mesial temporal cortex and right thalamus. Functional couplings between these brain regions were abnormally enhanced in the left-sided epilepsy patients. The right thalamic left superior temporal coupling was also abnormally enhanced in the right-sided epilepsy patients, but differentially from that in the left-sided patients. The other principal component was characterized by a pattern of metabolic interactions involving right and left mid prefrontal and right superior temporal cortex. Although both the right- and left-sided epilepsy patients showed decreased functional couplings between left mid prefrontal and the other brain regions, a weaker right-left mid prefrontal coupling in the left-sided epilepsy patients best distinguished them from the right-sided patients. The two mutually independent, abnormal metabolic patterns each predicted verbal intelligence deficits in the patients. The findings suggest a site-dependent reorganization of two independent, language-subserving pathways in temporal lobe epilepsy. PMID- 10390025 TI - Genetically dissociated components of working memory: evidence from Down's and Williams syndrome. AB - Wang and Bellugi [J clin exp Neuropsychol 1994;16:317 22] have suggested that Down's and Williams syndrome might be associated with specific and contrasting working memory deficits; with impaired verbal short-term memory in Down's syndrome, and a visuo-spatial short-term memory deficit in Williams syndrome. In two studies we examine whether these apparent deficits might simply be a consequence of the general pattern of learning difficulties associated with these disorders. Experiment 1 compared verbal and visuo-spatial short-term memory abilities in these groups, using analysis of covariance to control for mental age differences. In Experiment 2 individuals with Williams syndrome were matched to control groups for non-verbal mental age, and the short-term memory abilities of these matched groups were compared. The results of both experiments are broadly consistent with those reported by Wang and Bellugi, and support the view that working memory can be dissociated into separate subsystems. PMID- 10390026 TI - Vestibular stimulation affects dichotic lexical decision performance. AB - We report an experimental attempt to shift, by vestibular stimulation, healthy subjects' right ear advantage (REA) in a dichotic listening (DL) task with words and nonwords as stimuli. Forty right-handed men performed the task under two different conditions, once while sitting in a stationary turning chair (baseline) and once during sinusoidal rotation. In this latter condition, every other stimulation was received during maximal left-to-right (i.e., clockwise), every other during maximal right-to-left (i.e., counterclockwise) acceleration. There was a reliable REA for lexical decision accuracy in the baseline and right-to left trials but not during left-to-right rotation. While right ear performance was unaffected by rotation, there were more correct lexical decisions to left ear targets exclusively during left-to-right turns (one-tailed P = 0.05). Since there were no parallel shifts in auditory thresholds under the different conditions, this effect is not due to any hypothetical auditory-vestibular interactions on a primary sensory level. The improvement in left ear DL performance, although small in our study, is comparable to the symptom-alleviating effect of caloric vestibular stimulation in patients with left-sided hemispatial neglect and interpreted as a consequence of a rotation-induced attentional shift towards the left hemispace. PMID- 10390027 TI - The absent mind: further investigations of sustained attention to response. AB - We have previously demonstrated that performance on a brief and conceptually simple laboratory task (the Sustained Attention to Response Test: SART) was predictive of everyday attentional failures and action slips in brain injured patients and normal control participants. The SART is a go-no-go paradigm in which the no-go target appears rarely and unpredictably. Performance on this measure was previously interpreted as requiring sustained attention to response rather than a putative 'response inhibition' capacity. Three further studies are presented which support this claim. They demonstrate that performance is crucially determined by the duration of time over which attention must be maintained on one's own actions that this demand underpins the task's relationship to everyday attentional lapses. In keeping with a number of recent studies it suggests that inefficiencies in the maintenance of attentional control may be apparent over much briefer periods than is traditionally considered using vigilance measures and analysis. PMID- 10390028 TI - A neural basis for category and modality specificity of semantic knowledge. AB - Prevalent theories hold that semantic memory is organized by sensorimotor modality (e.g., visual knowledge, motor knowledge). While some neuroimaging studies support this idea, it cannot account for the category specific (e.g., living things) knowledge impairments seen in some brain damaged patients that cut across modalities. In this article we test an alternative model of how damage to interactive, modality-specific neural regions might give rise to these categorical impairments. Functional MRI was used to examine a cortical area with a known modality-specific function during the retrieval of visual and non-visual knowledge about living and non-living things. The specific predictions of our model regarding the signal observed in this area were confirmed, supporting the notion that semantic memory is functionally segregated into anatomically discrete, but highly interactive, modality-specific regions. PMID- 10390029 TI - Effects of experimentally-induced emotional states on frontal lobe cognitive task performance. AB - A growing body of evidence suggests that dysphoric and euphoric emotional states are associated with reliable patterns of frontal lobe activity. Specifically, dysphoric affect coincides with greater right than left frontal lobe activity, and euphoric affect tends to correspond with a converse pattern of activity. The present study examined whether cognitive outcomes associated with the left and right frontal lobes are differentially influenced by dysphoric and euphoric affect. In a completely between-groups design, 60 dextral women were administered either the positive or negative conditions of the Velten Mood Induction Procedure, and they subsequently completed either a verbal or figural fluency test. Euphoria resulted in better verbal than figural fluency performance, and dysphoria yielded better figural than verbal fluency outcomes. These findings are consistent with electrophysiological data concerning frontal lobe activity during euphoric and dysphoric affect, and they underscore the notion that affective influences upon cognition are more complicated than previously thought. PMID- 10390030 TI - Parkinson's disease and the control of size and speed in handwriting. AB - This experiment investigated whether Parkinson's disease (PD) patients experience problems in producing stroke size, stroke duration or both, in a handwriting task. Thirteen PD patients and 15 elderly controls wrote four patterns of varying complexity on a digitizer tablet. The participants were instructed to execute the writing movements: at a normal size and speed; as fast as possible; two times larger than normal; and two times larger and as fast as possible. PD patients had no difficulty increasing speed while maintaining size and had no difficulty increasing size while maintaining speed. However, they showed significantly smaller size increases in the two times larger condition as compared to the elderly controls. The conditions were also simulated by a neural network model of normal and PD movement control that produced a stroke pattern that approximated the experimental data. For the instructions used, the results suggest that when patients scale speed, they have no difficulty controlling force amplitude, but when they scale stroke size, they have a problem controlling force amplitude. Thus, PD patients may have reduced capability to maintain a given force level for the stroke time periods tested with the instructions. PMID- 10390031 TI - Spatial influences on motor and language function. AB - In subjects with parietal lobe lesions, performance on motor and language tasks differed as a function of the side of space to which subjects directed their attention or acted. Subjects with left parietal lesions performed better when attention was directed to stimuli in left hemispace (that is, the left side of their environment), and those with right parietal lesions showed a similar effect when attending to stimuli in right hemispace. Hemispace effects were not observed in subjects with lesions located elsewhere in the cerebral hemispheres, or in subjects with subcortical lesions. These data are consistent with the view that not only motor but also cognitive operations such as language, which do not appear to have any intrinsic spatial organization, are maintained in registration with spatial systems, and that this attention-requiring linkage confers a processing advantage. PMID- 10390032 TI - Motor and perceptual factors in pseudoneglect. AB - An important variant of the traditional line bisection task has involved a mechanical device invented by Bisiach and his colleagues (Bisiach et al. Perceptual and premotor factors of unilateral neglect. Neurology 1990;40:1278-81 [3]). This tool was devised to dissociate motor from perceptual factors in hemi spatial neglect, by means of a mid-line indicator which moved 'congruently' or 'non-congruently' with the direction of hand movement. In the non-congruent condition, Bisiach was able to demonstrate a reduction, or reversal, of the direction of bisection error in a number of patients with neglect. These errors were interpreted as instances of 'motor' neglect. Bisiach et al. [3] also tested 10 normal subjects, who did not differ on the two conditions of the task. However, the original experiment [3] required the use of the right hand only, and it has since become clear that bisection errors in normal subjects (i.e. pseudoneglect) are more substantial when dextral subjects use their left hands. By using a modified version of the Bisiach Tool we show that there is an effect of the motor versus perceptual condition on this task, but only when subjects use their non-dominant (left) hand. PMID- 10390033 TI - Vestibulo-ocular dysfunction induced by cortical damage in man: a case report. AB - We studied the rare case of a patient presenting with vestibulo-ocular dysfunction and clinical vestibular symptoms after right temporo-parietal cortex infarction. The vestibulo-ocular reflex (VOR) was elicited in the dark, by sinusoidal (0.02; 0.05 and 0.1 Hz) and by step velocity rotation (100 degrees/s2) in clockwise and counterclockwise directions. Horizontal and vertical eye movements were recorded by DC electro-oculography (EOG). When compared to a control group of 8 healthy subjects, this patient presented VOR asymmetry with (1) a significant VOR velocity bias toward the lesioned side revealed as a vestibulo-ocular offset that occurred only under dynamic conditions (2) a significant reduction of the VOR time constant when rotation was directed to the lesioned side. VOR gain was normal. We suggest that the parieto-temporal cortex is implicated in the regulation of vestibulo-ocular symmetry in man. This cortical processing of vestibular integration might involve a multidimensional velocity storage integrator that subserves the maintenance of spatial coordinates along the spatial vertical axis. PMID- 10390034 TI - How efficient are central mechanisms for the learning and retention of coincident timing actions? AB - We compared the adaptive strategy and retention capacity of a deafferented subject and control subjects when intercepting, with a sliding-throw, an apparent movement coming at various speeds. Subjects were submitted to five practice sessions (30 trials per session) and to a retention test. The throwing kinematics was analysed, and spatial and temporal performance errors were measured. With practice, the deafferented subject showed modifications in movement initiation strategies and throwing patterns. With a slow apparent movement, the deafferented subject's initial behavior was characterized by short movement initiation and movement times. With practice, she showed an important increase in movement time in session 5, allowing longer visual control and leading to better temporal and spatial accuracy than that shown in session 1. In the retention session, the deafferented patient showed a late movement initiation strategy, similar to that of the control subjects. This increased movement initiation time was accompanied by an improved temporal accuracy compared to the deafferented subject's early results. However, spatial accuracy improvement was labile and could not be maintained over the retention interval. At the fast speed, all temporal components of the response, namely, movement initiation time (MIT), movement time (MT), and disk travel time (DTT), were similar for the deafferented and control subjects. Overall, the deafferented subject reduced her temporal error through practice, though without attaining the control subjects' accuracy. However, with a fast-moving stimulus, she showed a deteriorated spatial accuracy, even doubling her spatial errors at retention. In brief, the deafferented subject achieved proper temporal (perceptivo-cognitive) lasting control of her interceptive action, whereas spatial (sensorimotor) regulation raised mnemonic problems. PMID- 10390035 TI - Facilitation and inhibition arising from the exogenous orienting of covert attention depends on the temporal properties of spatial cues and targets. AB - On the covert orienting of visual attention task (COVAT), responses to targets appearing at the location indicated by a non-predictive spatial cue are faster than responses to targets appearing at uncued locations when stimulus onset asynchrony (SOA) is less than approximately 200 ms. For longer SOAs, this pattern reverses and RTs to targets appearing at uncued locations become faster than RTs to targets appearing at the cued location. This facilitation followed by inhibition has been termed the biphasic effect of non-predictive peripheral spatial cues. Currently, there is debate about whether these two processes are independent. This issue was addressed in a series of experiments where the temporal overlap between the peripheral cue and target was manipulated at both short and long SOAs. Results showed that facilitation was present only when the SOA was short and there was temporal overlap between cue and target. Conversely, inhibition occurred only when the SOA was long and there was no temporal overlap between cue and target. The biphasic effect, with an early facilitation followed by a later inhibition, occurred only when the cue duration was fixed such that there was temporal overlap between the cue and target at short but not long SOAs. In a final experiment, the duration of targets the temporal overlap between cue and target and the SOA were manipulated factorially. The results showed that facilitation occurred only when the SOA was short, there was temporal overlap between cue and target and the target remained visible until the subject responded. These results suggest that the facilitation and inhibition found on COVATs which use non-informative peripheral cues are independent processes and their presence and magnitude is related to the temporal properties of cues and targets. PMID- 10390036 TI - Residual perceptual distortion in 'recovered' hemispatial neglect. AB - In most neglect patients, line bisection errors become smaller on repeated tests over the months following the lesion. We have tried to determine in two typical patients whether this is because of a real reduction in the perceptual distortions that appear to underlie line bisection errors in neglect, or whether it reflects a learned behavioural strategy to counteract those perceptual biases. We tested the patients on two occasions (2 and 12 months post-stroke), on line bisection and also on the 'Landmark' task. The data indicated that at the first testing session both patients showed strong 'perceptual' neglect, making large rightward errors in the standard bisection task and uniformly leftward pointing in the Landmark task. On the second occasion, as expected, both patients showed a marked recovery when tested with the line bisection task, making only very small errors. In contrast, their landmark performance was still markedly biased in the same direction as before. These findings suggest that despite their apparent recovery on the bisection task, both patients still experience some form of perceptual distortion of horizontal lines. It is suggested that the Landmark task may provide a sensitive means for identifying real recovery of the underlying perceptual deficit. PMID- 10390038 TI - The disability paradox: high quality of life against all odds. AB - This paper builds on the work of Sol Levine to examine a disability paradox: Why do many people with serious and persistent disabilities report that they experience a good or excellent quality of life when to most external observers these individuals seem to live an undesirable daily existence? The paper uses a qualitative approach to develop an explanation of this paradox using semi structured interviews with 153 persons with disabilities. 54.3% of the respondents with moderate to serious disabilities reported having an excellent or good quality of life confirming the existence of the disability paradox. Analysis of the interviews reveals that for both those who report that they have a good and those who say they have a poor quality of life, quality of life is dependent upon finding a balance between body, mind and spirit in the self and on establishing and maintaining an harmonious set of relationships within the person's social context and external environment. A theoretical framework is developed to express these relationships. The findings are discussed for those with and without disabilities and directions are given for future research. PMID- 10390037 TI - Impact of the surface slipperiness of grasped objects on their subsequent acceleration. AB - Seven subjects were asked to reach and grasp an object between the thumb and index finger, lift it about 30 cm high and 25 cm forward from one table to another, at their preferred speed. The perpendicular grip force and the tangential load force applied to the contact surface were digitized at 500 Hz and stored on a laboratory computer. The trajectory of the wrist and of the object was recorded using four infrared cameras tracking the movement of reflective markers attached to the distal styloid process of the radius and on the top of the object. The aim of this study was to demonstrate the influence of low friction (i.e. surface slipperiness) on the acceleration of the wrist. Friction was reduced by coating the smooth brass grasping surface with talc. The seven subjects had skin to surface coefficients of friction which ranged from 0.52-1.18 for dry brass and 0.24 0.34 for talc-coated brass. Two weights (418 and 1070 g) were used with each surface. The results indicated that with the slippery surface the necessary higher grip force/load force ratio was produced by an increase in the grip force and by a decrease in the wrist acceleration and a consequent reduction in the load force. This strategy was observed for both weights over a range of grip strengths between 21-98% of the individual's maximum voluntary contraction (MVC). This implies that even with adequate grip force reserves the reduction in acceleration is an acceptable and probable alternative solution to the force control problem. Our results also suggested that the loading rate and the object acceleration were planned and controlled together which emphasizes the role played by a predictive mechanism in organizing the kinematics of movements involving hand-held objects. This study shows that friction of the grasping surface not only affects the prehensile force dynamics, but it also influences the kinematics of the entire upper limb. PMID- 10390039 TI - Patient satisfaction in developing countries. AB - Efforts to obtain useful information on patient satisfaction in Indonesia have been frustrated by a tendency of respondents to withhold critical comment. A survey of 75 patients in eleven health centers on three islands attempted to obtain credible information on satisfaction by asking for information on events, not opinions, and on the relative importance of the factors surveyed. Unlike previous research where 95% of respondents typically answered they were 'fully satisfied', 28% of the respondents replied that their consultation had not been conducted in private (ranked first in importance among the nonmedical factors), 65% said the facility could be cleaner (ranked second in importance) and 19-48% reported not receiving various kinds of information (ranked third). Lending credence to these results, the respondents were able to support their positive answers with corroborative information in a high percentage of instances. The ranking of relative importance of satisfaction factors was unexpected. At the bottom of the list were continuity of provider, waiting time, availability of amenities, cost and social interaction with the provider. Despite the diversity among cultures that is characteristic of Indonesia, there was a high degree of similarity in the importance rankings among respondents on the three islands which were chosen to represent cultural divisions in the nation. The only notable differences in the rankings of relative importance appeared to be a function of the purpose of the visit to the facility. PMID- 10390040 TI - Risk analysis of poor health and growth failure of children in the central highlands of Guatemala. AB - Child morbidity and growth failure are multidimensional phenomena. An assessment was undertaken of the food and nonfood risk factors of poor health and growth failure in children of different age groups in the central highlands of Guatemala. The aim was to identify high risk factors in under-five and school-age children. Under-five children at high risk of being ill tended to come from households with: high needs of child care, a lack of access to a private well or piped water, and no sewage connection. Women's illiteracy constituted an additional risk factor for diarrheal disease in under-five children. Growth failure in under-five children was mainly due to chronic factors: 74% were stunted, 6% were wasted and 44% were weight deficient. These prevalence rates were lower among school-age children. Low per capita food availability, and particularly the absence in the household of self-produced staple foods, was the most significant risk factor of growth failure in under-five children, followed by high risk of being ill, and participation by women in farm production. The latter was particularly a risk factor of wasting. Nonfood risk factors were most important for growth failure in school-age children. These factors included: sanitation, housing conditions, women's literacy status, and adult women's body mass index. Participation in farm production by school-age children was associated with a higher risk of growth failure in younger siblings. It is concluded that multisectoral programs need to reduce the impact of various risk factors of poor health and growth failure in children, and be careful not to introduce new risk factors. Depending on which age group is targeted, such programs should either prioritize improvements in household food availability, or nonfood interventions that reduce women's illiteracy and improve sanitary and housing conditions. PMID- 10390041 TI - Predictors of childhood immunization completion in a rural population. AB - Despite the availability of effective vaccines, immunization rates among two-year old children continue to be low in many areas of the United States including rural West Virginia. The goal of this study was to identify barriers to childhood immunization in rural West Virginia and determine factors that were important in the completion of the childhood immunization schedule. A telephone survey was used to collect data from a randomly selected sample of 316 mothers, of two-year olds, from 18 rural counties of West Virginia. Results indicated that two-thirds or 65% of the children in the study sample had completed their recommended immunizations by two years of age. Immunization barriers identified in this study include: living in health professional shortage areas, lack of health insurance, negative beliefs and attitudes regarding childhood immunizations, problems accessing the immunization clinic, and a perception of inadequate support from the immunization clinic. Results of the structural equation modeling, using LISREL-8, indicated that 20% of the variation in immunization completion (R2 = 0.197) was explained by attitude towards immunization and perceived support received from the immunization clinic. Furthermore, 42% of the variation in attitude towards immunization (R2 = 0.419) was explained by immunization-related beliefs, and 28% of the variation in immunization-related beliefs (the R2 = 0.277) was explained by general problems faced during immunization and perceived clinic support. The study concluded that positive immunization-related beliefs and attitudes, support from the immunization clinic, and ease of the immunization seeking process are important factors in the timely completion of the childhood immunization schedule. PMID- 10390042 TI - Starting mental health services in Cambodia. AB - Cambodia has undergone massive psychosocial trauma in the last few decades, but has had virtually no western-style mental health services. For the first time in Cambodia a number of mental health clinics in rural areas have been started. This experience is used to discuss the risks and opportunities in introducing these services in the present war-torn situation. Basic statistics from the clinics are presented in the context of the historical and traditional setting, and the effort to maintain a culturally informed approach is described. The contrasting results in the clinics are analyzed in relation to factors intrinsic to the health care system and those related to the local population in order to highlight the issues involved in establishing future mental health services, both locally in other provinces and in situations similar to Cambodia. The efficacy of introducing low-cost, basic mental health care is shown, and related to the need to find solutions for prevailing problems on the psychosocial level. They can be introduced with modest means, and can be complementary to local health beliefs and traditional healing. In introducing mental health services, an approach is needed which adapts to the absorption potential of the health system as well as to the patients' need to find meaningful help. Existing resources, from the traditional healing sector to rudimentary village structures, cannot be neglected in the rehabilitation of the community, or in interventions to help the individual patient. PMID- 10390043 TI - From African health and influenza pandemics to disease in Russia: the medical history of K. David Patterson. AB - This paper is a personal commentary on the contributions of noted medical historian K. David Patterson. The eightieth anniversary of the onset of the Great Influenza Pandemic of 1918-19 serves as a reminder of his contributions. His works on slavery and disease in Africa, influenza diffusion and the history of cholera and other diseases periodically appeared in Social Science and Medicine. With a publishing career that lasted from 1971 to 1996, his enduring contributions include revised estimates of mortality in Africa during the influenza pandemic of 1918-1919. PMID- 10390044 TI - Attaining health for all through community partnerships: principles of the census based, impact-oriented (CBIO) approach to primary health care developed in Bolivia, South America. AB - This article describes a flexible primary health care methodology which was developed by Andean Rural Health Care and its colleagues in Bolivia, South America. This methodology, the census-based, impact-oriented (CBIO) approach to primary health care, involves determining local health priorities as defined both by locally acquired epidemiologic information and by the local people themselves. The CBIO approach to primary health care is now functioning successfully at seven program sites in Bolivia, which together serve 75,000 people in urban and rural communities in three distinct cultural and ecological regions of the country. High levels of coverage of basic health services can be achieved through a system of 'epidemographic' surveillance of all families and through home delivery, when needed, of priority services to those at risk. When the services provided are based on local health priorities, when they are provided in a technically effective manner, and when the community has a strong partnership in planning, implementation and evaluation, then the CBIO approach to primary health care will lead to measurable health improvements as defined by changes in population-based rates of mortality and illness in the community. On the basis of our experience, we believe that the CBIO approach offers great potential for strengthening the effectiveness of local health programs in impoverished communities around the world in a way which fosters community ownership and, hence, long-term sustainability. PMID- 10390045 TI - Perceptions of soil-eating and anaemia among pregnant women on the Kenyan coast. AB - After a clinical study at Kilifi District hospital had shown a high prevalence of geophagy among pregnant women, and a strong association of geophagy, anaemia and iron depletion, 52 pregnant women from the same hospital, and 4 traditional healers from the surroundings of Kilifi in Kenya were interviewed on the topic of soil-eating and its perceived causes and consequences. The findings were substantiated by results from an earlier anthropological study on maternal health and anaemia in the same study area. Most of the pregnant women (73%) ate soil regularly. They mainly ate the soil from walls of houses, and their estimated median daily ingestion was 41.5 g. They described soil-eating as a predominantly female practice with strong relations to fertility and reproduction. They made associations between soil-eating, the condition of the blood and certain bodily states: pregnancy, lack of blood (upungufu wa damu), an illness called safura involving "weak" blood, and worms (minyolo). The relationships the women described between soil-eating and illness resemble to some extent the causalities explored in biomedical research on soil-eating, anaemia and intestinal worm infections. However the women did not conceptualise the issue in terms of the single causal links characteristic of most scientific thought. Instead, they acknowledged the existence of multiple links between phenomena which they observed in their own and other women's bodies. The women's ideas about soil eating and their bodies shows the significance of both social and cultural context on the ways in which women derive knowledge from, and make sense of their bodily states. The cultural associations of soil-eating with blood, fertility and femininity exist alongside knowledge of its links to illness. Our findings show that soil-eating is more than just a physiologically induced behaviour; it is a rich cultural practice. PMID- 10390046 TI - The social ecology of syphilis. AB - Factors affecting the transmission of syphilis can be categorized into those acting at the level of individuals (e.g., number of sex partners) and others at the level of the sociophysical environment (e.g., availability of treatment services for curable infections). In a prior study, we identified several sociophysical factors correlated with the ten-year mean syphilis rate in a regression analysis of United States counties. In the present study we used qualitative methods to investigate additional aspects of some factors in the regression, as well as to identify entirely new factors. Twelve counties with populations less than 100,000 and ten-year mean syphilis rates that were greater or less than expected by the regression model were selected for a three to five day visit. The case study protocol included observations, unstructured interviews with care providers and county residents, and a standardized questionnaire completed by state and local sexually transmitted disease control personnel pertaining to characteristics and practices of the local health department. Comparisons of the field notes and questionnaires revealed patterns of factors of the sociophysical environment that potentially affect county syphilis rates. These included access to the health department STD clinic, race relations, employment opportunities for minorities, interagency coordination, STD outreach activities, the social acceptability of discussing STDs, and intercommunity dynamics. In addition we noted the disproportionate influence of particular individuals on these factors. Some of the factors identified are readily quantifiable and could enhance the predictive power of multivariable models of county syphilis rates. The hypotheses generated by this study may also lead to a better measurement and understanding of potentially important environmental determinants of community syphilis rates, and the development of new or enhanced prevention strategies. PMID- 10390047 TI - Implications of social class and race for urban public health policy making: a case study of HIV/AIDS and TB policy in Washington, DC. AB - This paper explores how social class and race affect the public health policy making process in an urban area. Ethnographic methods were used to collect and analyze information about HIV/AIDS and tuberculosis policy-making by the Washington, DC Commission of Public Health, Kingdon's conceptual model of policy making was used to analyze and understand the process. The problems of HIV/AIDS and tuberculosis in the district have important social class dimensions that were not always made explicit, but were instead defined in terms of 'race' and 'place'. Social class considerations and racial politics shaped what policies were developed or not developed and implemented successfully or failed. This study, which has national and international implications, concludes that there is a need to improve our understanding of the complex social dimensions of public health problems; there needs to be more consideration of the politics of strategy formulation and how issues of social class and race affect this process; and public health needs to strengthen its constituency in order to build support for the successful development and implementation of policy. PMID- 10390048 TI - Sexual regimes and sexual networking: the risk of an HIV/AIDS epidemic in Bangladesh. AB - Bangladesh adjoins the Asian region with the severest AIDS epidemic and has common borders with two of the most affected areas, the Indian Hill States and northern Burma. There has been disagreement about the danger to Bangladesh, one view citing the likelihood of transmission from neighbouring infected populations and the other claiming that the country's predominantly Muslim culture protects it. This paper reports on a 1995-1997 research project. Preliminary research was carried out in Dhaka in 1995-1996 which suggested that the poor squatter areas might well sustain an epidemic. The experience also showed that more accurate measures of sexual networking could be obtained from males than females. The 1997 field research reported here investigated 983 males, 52% single and 48% married in Chittagong city and two more rural areas of Chittagong Division in southeast Bangladesh. It was found that around half of all males and probably a somewhat lower proportion of females, experience premarital sexual relations, with males having a lower level of extramarital than premarital relations. The factor heightening Bangladesh's risk of an epidemic is that one-quarter of single males and a significant but lower level of married males have had relations with prostitutes. This is one explanation for quite high levels of STDs in Bangladesh. The factors restricting the chances of a major national epidemic are the small number of premarital sexual episodes per person and the low level of intravenous drug use. PMID- 10390049 TI - Reliability of perceived health by sex and age. AB - The aim of this study was to assess the test-retest reliability of a measure of perceived general health by sex and age. The study analyzed data from the nationally representative Mini-Finland Health Survey of 8000 adults aged 30 and over. The subjects were invited to attend a personal health interview and a health examination in 1978-1980. Altogether 7217 persons participated. Perceived general health was measured at the personal health interview and in the self administered questionnaire 1-6 weeks apart. The identical questions were: how would you assess your current health? The response alternatives were good, fairly good, intermediate, fairly poor, poor and cannot say. This study showed that among men and women unweighted agreement of the 'good-intermediate-poor' categorization of perceived health was around 70% and unweighted kappa-values were around 0.5. Only in the oldest age-group (75+) reliability declined below these levels. The fair or good reliability of perceived health observed in this study gives additional confidence for using this general measure of overall health status in future research. PMID- 10390050 TI - Susceptibility to oxidation of copper-induced plasma lipoproteins from Japanese eel: protective effect of vitellogenin on the oxidation of very low density lipoprotein. AB - The susceptibility to oxidation of copper-induced plasma lipoproteins from Japanese eel Anguilla japonica was examined with the guidance of thiobarbituric acid-reactive substances (TBARS). The TBARS values of copper-induced plasma lipoproteins increased with increasing the lipid-to-apolipoprotein ratios and very low density lipoprotein (VLDL) exhibited the highest TBARS value. On the other hand, vitellogenin, estrogen-induced precursor of egg yolk proteins, was resistant to copper-induced oxidation and seemed to chelate low concentrations of copper ion. Vitellogenin also protected the copper-induced oxidation of VLDL because of its antioxidant function. Vitellogenin seemed to serve as transition metals-binding lipoprotein by which free-radical reactions in the oocytes were extensively depressed. PMID- 10390051 TI - Identification of a pathogen-binding lectin in salmon serum. AB - A mannose-binding lectin was isolated from the blood serum of Atlantic salmon (Salmo salar). Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing and non-reducing conditions revealed a multimeric structure composed of 17000 Mr subunits. Hexosamine analysis and glycosidase digestion showed that the lectin is not glycosylated and amino acid analysis revealed no unusual compositional features. Using ruthenium red staining, the lectin was shown to bind Ca2+ ions. N-terminal sequencing by Edman degradation gave: H2N-TGAKGAEEGVVPAETRNQXPTGWFQFGS. A database search revealed no similarity to protein sequences deposited to date. Binding experiments using biotinylated lectin revealed that it specifically recognizes and binds to mannose on the surfaces of two salmon pathogens, Vibrio anguillarum and Aeromonas salmonicida, implying an immunological role for this lectin in Atlantic salmon. PMID- 10390052 TI - Dehydroepiandrosterone and related steroids alter 3T3-L1 preadipocyte proliferation and differentiation. AB - The purpose of the present study was to determine if the anti-adipogenic effects of dehydroepiandrosterone (DHEA) are mediated solely by DHEA or by one or more of its downstream metabolites. In Experiment 1, preconfluent proliferating cultures of 3T3-L1 preadipocytes were incubated for either 24 or 72 h with 0, 1, 5 or 25 microM DHEA, DHEA sulfate (DHEAS), testosterone, estrone and 17beta-estradiol. Pregnenolone, a precursor of DHEA(S), was also tested at these concentrations. After 24 h of incubation, DHEAS, 17beta-estradiol and estrone at the 1 microM level stimulated preadipocyte proliferation. In contrast, DHEA and 17beta estradiol at the 25 microM level attenuated proliferation to a greater extent than all other steroids. After 72 h of incubation, DHEA and 17beta-estradiol at the 25 microM level attenuated proliferation to a greater extent than all other steroids. In Experiment 2, post-confluent cultures of differentiating 3T3-L1 preadipocytes were incubated for 6 days with 0, 5, 30, or 60 microM levels of these steroids. Preadipocyte differentiation, as assessed by lipid content and glycerol-3-phosphate dehydrogenase activity, decreased markedly when treated with 30 and 60 microM DHEA, 17beta-estradiol, estrone and pregnenolone. In contrast, DHEAS had no impact on preadipocyte proliferation or differentiation. These results suggest that the anti-adipogenic actions of DHEA in adipose tissue may be mediated, in part, by one or more of its distal metabolites, including 17beta estradiol. PMID- 10390053 TI - Effects of nickel chloride on reproduction of the rat and possible antagonistic role of selenium. AB - Nickel (10-100 ppm added as NiCl2) was studied to determine its effects on reproduction of Wistar rats. In nine experimental groups, females, males or both were exposed to nickel in drinking water. In one female group and one male group, the drinking water was also supplemented with 0.3 ppm selenium (added as Na2SeO3). Breeding success and the growth and viability of pups were recorded. Nickel, copper and zinc concentrations in kidneys, liver and skin (with fur) of the females, males and pups were determined with an atomic absorption spectrophotometer. In addition, histology of the male testes (from control and nickel-exposed groups) was studied. The female exposures started 14, 28 or 100 days before copulation and continued during pregnancy and lactation. When the males were exposed (for 28 or 42 days before copulation), NiCl2 reduced both the number of pregnancies and the number of pups born. In the testes, NiCl2 induced shrinkage of the seminiferous tubules, which seemed to close some of the tubules. In the tubules, NiCl2 decreased the number of basal spermatogonia. When the females or both parents were exposed to NiCl2, pup mortality during lactation was high. However, when the females were drinking NiCl2 supplemented with selenium, all the pups survived and development of the total mass of the litters was even better than in the control group. In the same way, in males, selenium supplementation of the drinking water protected those pups that were born; but fertility was lower than with the control treatment. In the tissues studied, nickel accumulated most in the kidneys and then in the liver and skin. In each type of organ, there was a clear dose response relationship. In the pups, in particular, selenium (given to the females) increased the amount of nickel in tissues compared with corresponding administration of nickel without selenium. In summary, selenium seemed to counteract the deleterious effects of NiCl2 on the reproduction of rats. PMID- 10390054 TI - Cadmium induced thyroid dysfunction in chicken: hepatic type I iodothyronine 5' monodeiodinase activity and role of lipid peroxidation. AB - Administration of cadmium chloride (2.5 mg/kg body weight/day) to chickens daily for 15 days decreased serum triiodothyronine (T3) concentration (by 68.75%) without altering the levels of serum thyroxine (T4). Hepatic 5'-monodeiodinase (5'D-I) and superoxide dismutase (SOD) activities were also decreased (by 90.47% and 20.81% respectively) with a concomitant increase in lipid peroxidation (LPO, by 206.25%). Administration of the antioxidant vitamin E (alpha-tocopherol, 5 mg/kg body weight on alternate days) to cadmium intoxicated chickens restored thyroid function by maintaining normal hepatic 5'D-I activity and serum thyroid hormone concentrations. It also prevented cadmium-induced increase in LPO. We conclude that the metal-induced inhibition in hepatic 5'D-I activity is mediated through LPO. PMID- 10390055 TI - Tissue folate binding protein levels in transgenic mice with tumors and in non transgenic controls. AB - Localized folate deficiency may be a risk factor for cancer. Since, folate binding proteins (FBP) and reduced folate carrier proteins (RFC) mediate cellular transport of folate, we compared FBP concentrations in several organs from tumor bearing transgenic (TBT) mice and tumor-free non-transgenic controls (NTC) of the same strain, age, and fed identical diets. Liver, spleen, brain, small intestine and kidney were individually homogenized in phosphate-buffered saline (PBS) and separated into membrane, cytoplasmic, mitochondrial/lysomal and nuclear fractions (confirmed with marker enzymes). Homogenates and fractions was analyzed for total protein, and FBP. We used rabbit anti-bovine milk antibody and ELISA to measure FBP. FBP concentrations in kidney, small intestine, and spleen of TBT mice were higher than those of NTC mice; the opposite was true in liver and lung. FBP seemed to be upregulated in kidneys (all fractions), small intestine (all fractions), and spleen (cytoplasmic and nuclear fractions only) of TBT mice compared to NTC mice; the opposite appeared true in liver (all fractions) and lung (all fractions). FBP concentrations in brain, heart, and muscle of TBT mice were not different from those in brain, heart and muscle of NTC mice. A longitudinal study will determine if these changes in FBP concentrations precede tumor onset. PMID- 10390057 TI - Cadmium turnover in the hemocytes of Mercenaria mercenaria (L.) in relation to hemocyte turnover. AB - The turnover of cadmium (Cd) in the hemocytes of the quahog Mercenaria mercenaria L. was investigated using 109Cd radiolabeling, and compared to the rate of hemocyte turnover as estimated following [3H]thymidine incorporation. Quahogs were injected with 5 microl of 0.8 microM 109Cd, and the hemocytes sampled over a 60-day depuration period. Additional organisms were injected with 15 microl of 4.7 microM [3H]thymidine and radioactivity monitored over a 60-day period. Assuming that the loss of 3H from the hemocytes during the first 27 days of the experiment is due to hemocyte turnover, results indicate that Cd turnover rate (t 0.5 = 25.3 +/- 6.3 days) was similar to hemocyte turnover rate (t 0.5 = 28.5 +/- 14.3 days). This suggests that hemocytes retain Cd internally throughout their circulatory lifespan. PMID- 10390056 TI - Enhancement of anti-Aeromonas salmonicida activity in Atlantic salmon (Salmo salar) macrophages by a mannose-binding lectin. AB - We investigated the effects of a calcium-dependent mannose-binding lectin isolated from the serum of Atlantic salmon on Aeromonas salmonicida viability and the anti-A. salmonicida activity of Atlantic salmon macrophages. In the absence of other factors, binding of this lectin at concentrations of 0.8, 4.0 and 20.0 ng ml(-1) to virulent A. salmonicida failed to significantly reduce (P> 0.05) cell viability. However, binding of the lectin to A. salmonicida did result in significant (P < or = 0.05) dose-dependent increases in phagocytosis, and bactericidal activity. Significant increases (P < or = 0.05) were also observed in phagocyte respiratory burst activity within the lectin concentration range of 4.0-20.0 ng ml(-1) but the stimulation was not dose dependent at these lectin concentrations. At the lowest lectin concentration tested (0.32 ng ml(-1)), a significant decrease (P < or = 0.05) in respiratory burst was observed. The structure and activity of this lectin are similar to that of mammalian mannose binding lectins, which are known to play a pivotal role in innate immunity. The presence of this lectin may be an important defense mechanism against Gram negative bacteria such as A. salmonicida. PMID- 10390058 TI - Hepatic microsomal enzyme activity in the koala and tammar wallaby: high 17beta hydroxysteroid oxidoreductase activity in koala liver microsomes. AB - We have studied the hepatic microsomal xenobiotic metabolising capacity of koala (Phascolarctos cinereus) and tammar wallaby (Macropus eugenii). Total cytochrome P450 content in hepatic microsomes from koala (0.87 +/- 0.18 nmol/mg protein, n = 4, mean (S.D.) and rat were comparable while tammar wallaby displayed reduced P450 content (0.24 +/- 0.04 nmol/mg protein). Associated microsomal activities (NADPH cytochrome P450 reductase, aminopyrine N-demethylation, aniline hydroxylation, and androstenedione 6beta- and 16alpha-hydroxylation) in koala liver were similar to or reduced relative to rat. Hepatic microsomal NADPH supported 17beta-hydroxysteroid oxidoreductase (17beta-HSOR) activity was significantly higher in koala (9.99+/-3.08 nmol/mg protein/min) than in tammar wallaby liver (0.86 +/- 0.16 nmol/mg protein/min). However, when NADH was utilised as cofactor the activity was similar in both marsupial species (koala, 1.44 +/- 0.84 nmol/mg protein/min; tammar wallaby, 1.52 +/- 0.44 nmol/mg protein/min). Michaelis-Menten parameters for the kinetics of 17beta-HSOR androstenedione reduction by NADPH and NADH were determined in the koala. The Km for androstenedione was of the order of 1.9-4 microM (n = 4) irrespective of the cofactor used, whilst the Km for NADPH was 0.04-0.05 microM (n = 2) and for NADH was 134-430 microM (n = 2). Potential inhibitors were evaluated for their effects on NADPH-mediated 17beta-HSOR activity with menadione and, to lesser extents, menthone, benzaldehyde and metyrapone eliciting significant inhibition. From detailed kinetic studies menthone was found to be an uncompetitive inhibitor of the activity in koala liver (Ki 220 microM). PMID- 10390059 TI - Acute and chronic effects of toxic metals on viability, encystment and bioluminescence in the dinoflagellate Gonyaulax polyedra. AB - Toxicity bioassays based on survival were carried out with cells of the marine dinoflagellate Gonyaulax polyedra exposed to mercury (Hg2+ ), cadmium (Cd2+), lead (Pb2+) and copper (Cu2+). The toxicity scale of these metals found was Hg2+ > Cu2+ > Cd2+ > Pb2+. Cells exposed to metals promptly underwent encystment, which is an important strategy for surviving metal exposure. Following 48 h exposure to Cu2+, complete excystment occurred within 96 h after reinoculation of cells in fresh metal-free media, and with Pb2+ partial recovery occurred in that time. Bioluminescence was affected by the metals in a dose-dependent manner primarily by increasing the frequency of flashing, but the glow emission was also altered with acute Cu2+ and Pb2+ treatments. Several physiological processes in G. polyedra are under circadian control. Chronic exposures to metals caused no substantial alterations in the circadian rhythm of bioluminescence glow, indicating that the biological clock of this dinoflagellate is not sensitive to these metals at the concentrations tested. PMID- 10390060 TI - Extracellular and intracellular actions of azadirachtin on the electrophysiological properties of cultured rat DRG neurones. AB - The distinct electrophysiological actions of extracellular and intracellular azadirachtin (type A) were investigated using cultured dorsal root ganglion neurones from neonatal rats and the whole cell variant of the patch clamp technique. The cultured mammalian neurones were relatively insensitive to azadirachtin compared with some invertebrate preparations, such as insect chemosensory systems. Insect preparations have been found to respond to 1-100 nM azadirachtin. However, at concentrations of azadirachtin between 10 and 100 microM significant changes in the electrophysiological properties of cultured DRG neurones were seen. Extracellular application of azadirachtin prolonged the late repolarization phase of evoked action potentials and consistent with this produced modulation of voltage-activated K+ currents. This modulation involved an initial transient increase in K+ current followed by reversible inhibition when azadirachtin was applied at a concentration of 10 microM. Higher concentrations of azadirachtin (50 and 100 microM), had only inhibitory effects on voltage activated K+ currents. Azadirachtin produced a degree of voltage-dependent inhibition, with a greater level of K+ current inhibition observed when neurones were held at -90 mV compared with -40 mV. Prolonged application of azadirachtin for 24 h reversibly reduced action potential amplitude. In contrast intracellular application of 100 microM azadirachtin via the patch pipette solution had no significant effects on action potentials but produced an increase in conductance. Intracellular azadirachtin activated several conductances including a TEA sensitive K + current and an inward current that had an estimated reversal potential close to 0 mV. In conclusion, data showed that azadirachtin had different effects when it was applied inside and outside the neurones and that azadirachtin at high concentrations reversibly altered neuronal excitability mainly by modulating potassium conductances. It appears that rat cultured DRG neurones are less affected by azadirachtin than some insect sensory systems. PMID- 10390062 TI - Training in cervical cytology, past, present and future. PMID- 10390061 TI - Effect of a 14-day hindlimb suspension on beta-adrenoreceptors in rats. AB - Exposure to long-term simulated microgravity exhibits reduced sympathetic nervous system activity. This study tested the hypothesis that the hypersensitivity of adrenoreceptors would explain partly many other features of the hemodynamic consequences of return from space. The biochemical properties of the beta adrenoreceptors (betaAR) were determined using 125I-cyanopindolol (125I-CYP) binding in three rat groups: (1) The first experimental group consisted of 24 h restrained orthostatic rats in the horizontal position, to test the early effect of the attachment to the suspension device; (2) the second experimental group consisted of 24 h-restrained antiorthostatic rats, to test the early effect of the suspension; (3) the third experimental group consisted of 14 day-restrained antiorthostatic rats, to test the long term effect of the suspension. The study was performed in two organs involved in blood pressure regulation, i.e. the heart (atria and ventricles were separated) and kidneys. The Scatchard analysis of 125I cyanopindolol binding in both organs indicated no significant alterations in the dissociation constant (Kd) and the maximum binding capacity (Bmax) in the three experimental groups. These results do not allow the conclusion about the SNS adaptation pattern to simulated microgravity. Thus, the hypothesis that betaAR are involved in the cardiovascular adaptation to simulated microgravity is not verified in this model where, as a matter of fact, cardiovascular deconditioning is not verified even if this model is widely used. PMID- 10390063 TI - External quality assessment (EQA) in gynaecological cytology: radical rethinking required? PMID- 10390064 TI - Cervical intraepithelial neoplasia grade III (CIN III) and invasive cervical carcinoma: the yawning gap revisited and the treatment of risk. AB - In a 3-year study of the population of Southampton and south-west Hampshire there were 10 times as many cases of CIN III compared with invasive squamous carcinoma (700 compared with 70). The peak incidence of CIN III per 1000 screened women years was in those aged 25-29 years, which was 20 years earlier than the peak incidence of invasive cervical cancer per 1000 women years at risk. Ninety percent of CIN III was diagnosed in women under 50 years. There were 14 cases of cervical glandular intraepithelial neoplasia grade III (CGIN III), three coexisting with CIN III, all in women aged under 50 years: the gap between intraepithelial and invasive lesions was not seen for glandular neoplasia. Although referral was for at least moderate dyskaryosis in 86.8% of women with CIN III or CGIN III, most had been screened previously, either having had mild abnormalities requiring repeat cytology (39.8%) or negative cytology (34.5%). Only 12 women aged > or = 50 years had previous negative cytology: 21.4% compared with 35.6% of women aged < 50 years (P = 0.034). The results of this study suggest that the best opportunity for preventing invasive squamous cell carcinoma lies in screening women aged 20-39 years when the incidence of CIN III in the screened population is highest and before the peak incidence of invasive disease. The results also indicate the importance of repeated screening and follow up of minor cytological abnormalities in the detection of CIN III. The benefit of screening must be regarded as a treatment of risk, since it is almost certain that a high proportion of CIN III regresses or persists unchanged. PMID- 10390065 TI - Diagnostic value of qualitative and quantitative variables in thyroid lesions. AB - Two hundred and thirty-three thyroid lesions were studied by fine needle aspiration (FNA) cytology using standard cytologic criteria available in the literature. These included 114 cases of nodular colloid goitre (NCG), 47 cases of Hashimoto's thyroiditis (HT), 12 follicular adenomas (FAd), five cases of subacute thyroiditis and three cases of thyrotoxicosis among the benign lesions. The malignant lesions seen were 30 cases of papillary carcinoma (PCa), 16 follicular carcinomas (FCa), three cases with double lesions, e.g. papillary carcinoma with coexisting NCG, and three of papillary carcinoma with HT. Emphasis was given to eight qualitative and quantitative (morphometric) variables in these various thyroid lesions. Cell measurements were done using a Visopan Lux projection microscope. The three qualitative variables included type of nuclear membrane (regular/irregular), type of nuclear chromatin and the presence or absence of conspicuous nucleoli. The quantitative variables studied were nuclear diameter, nuclear area, cytoplasmic diameter, cytoplasmic area & N/C ratio. Statistical analysis was performed in order to know whether the standard cytologic criteria used at FNA cytology in the literature (increased cellularity, microfollicles, increased N/C ratio, absence of significant haemosiderin-laden macrophages and scanty colloid) could differentiate a follicular adenoma from a follicular carcinoma. A statistical analysis was also performed to establish the utility of the qualitative and quantitative variables. The results showed that none of the standard cytologic criteria applied could differentiate follicular adenoma from a follicular carcinoma. With regard to qualitative variables, irregularity of nuclear membrane and presence of conspicuous nucleoli were most significant in papillary carcinoma, followed by follicular carcinoma, then by follicular adenoma; these features being hardly evident in nodular colloid goitre and Hashimoto's thyroiditis. A coarse nuclear chromatin was most significant in follicular carcinomas followed by follicular adenomas. It was less obvious in the benign conditions, but more prominent in Hashimoto's thyroiditis compared with a goitre. It was also not obvious in a papillary carcinoma. Of the quantitative variables, all measurements were greatest in PCa > FCa > FAd > NCG = HT. When differentiating follicular adenoma from follicular carcinoma the qualitative variables of significance were the presence or absence of nucleoli, the chromatin pattern and regularity/irregularity of nuclear membrane; the nuclear area was the most important feature among quantitative variables. PMID- 10390067 TI - Difficulties in the fine needle aspiration (FNA) diagnosis of schwannoma. AB - The results of nine FNAs of eight histologically proven schwannomas are presented. In only one of the aspirates was a diagnosis of schwannoma made; three additional cases were diagnosed as 'spindle cell neoplasm'. Two of the cases were considered to be non-diagnostic due to hypocellularity, while two cases containing cellular material raised the differential diagnosis of granulation tissue and granulomatous inflammation due to the presence of epithelioid cells and an inflammatory infiltrate, and are illustrated here. The remaining FNA was felt suggestive of branchial cleft cyst due to the presence of only cystic fluid in a neck mass. We observe that: (i) the acquisition of an adequate, representative specimen via FNA is often difficult in schwannoma, and (ii) diagnostic difficulties may be encountered in cases in which cellular material is obtained. PMID- 10390066 TI - Cytological and immunocytochemical evaluation of thyroid and breast masses in patients with a previous neoplasm: case reports. AB - The diagnosis of secondary tumours represents one of the most important fields in the application of fine needle aspiration cytology (FNAC). We studied two patients, one with a history of breast cancer and one with a previous tumour of the thyroid, who showed a second mass, in the thyroid and in the breast, respectively, during follow up. The aim of our study was to evaluate if cytology, performed on FNAC smears, may distinguish a metastatic lesion from a second primary tumour, or if further immunocytochemistry should be performed. Our data demonstrate that, while cytology may be indicative of a second primary tumour, the histotype should be confirmed by immunocytochemical staining. PMID- 10390068 TI - Fine needle aspiration biopsy of the spleen in pyrexia of unknown origin. AB - To evaluate the diagnostic utility, value and potential risk of fine needle aspiration biopsy of spleen (sFNAB) in patients with splenomegaly in pyrexia of unknown origin (PUO), a retrospective analysis of medical records and cytological material of 31 patients on whom FNAB was performed between April 1994 and October 1997 was done. The patients were HIV- and presented with PUO. All other relevant investigations were negative. The spleen was either palpable or detected to have space-occupying lesions on ultrasonography (USG). The splenic aspirates showed tuberculosis in 11 patients (35.4%) and inconclusive or reactive changes in nine patients (25.8%). One case out of this group proved to be Kaposi's sarcoma on autopsy. The other diseases encountered were leishmaniasis (n = 3), non-Hodgkin's lymphoma (n = 4), fungal infections (n = 2), Hodgkin's lymphoma (n = 1). The patients who were diagnosed as having tuberculosis had epithelioid cells, giant cells, necrosis and inflammatory cells in various combinations. AFB positivity was 63.6%. The other cases which showed granulomas but no AFB were diagnosed on empirical grounds and all responded to the anti-tuberculosis therapy. No complications were encountered with the procedure. Therefore the authors conclude that sFNAB is rewarding in patients where all other non-invasive modalities of diagnosis have failed. PMID- 10390069 TI - A cytologist's view of China--Citizen Ambassador Program Cytopathology Delegation to the People's Republic of China, February 1997. PMID- 10390070 TI - Fine needle aspiration (FNA) in the management of palpable masses in Ibadan: impact on the cost of care. AB - Fine needle aspiration cytology is a relatively new technique in the management of palpable lesions in Ibadan. In the University College Hospital (UCH) Ibadan, as in most centres in Nigeria, inadequate facilities, a heavy patient load, financial constraints and an unreliable supply of basic necessities like water, often delay definitive diagnosis and management. In order to alleviate the patient's problems and provide prompt and accurate diagnosis, an FNA Cytology Clinic was set up in the Pathology Department, UCH, Ibadan, managed by the pathology team. This report represents the results of our experience. The cost effectiveness and impact on cost of care are highlighted. FNA costs N250.00 (pounds sterling 2.00), whilst cost of biopsy can vary from N5000.00 to N10000.00 (pounds sterling 35.00 to pounds sterling 70.00). A total of 752 satisfactory smears was reviewed during the 3-year period 1995-97 from various sites including breast (n = 295), lymph node (n = 183) and thyroid (n = 143). Diagnostic accuracy varied with different sites, the accuracy rate for breast, lymph node and thyroid malignancy being approximately 100%, 80% and 93%, respectively. PMID- 10390071 TI - The cytology of bronchial brushings from a malignant paraganglioma metastasizing to the lung. PMID- 10390072 TI - Diagnosis of hepatic metastasis of adenoid cystic carcinoma of the salivary gland by fine needle aspiration. PMID- 10390073 TI - Leydig cell tumour of the testis: cytological findings on fine needle aspiration. PMID- 10390074 TI - Acute myeloblastic leukaemia: graft-versus-host and graft-versus-leukaemia responses to autologous IL-2 activated lymphocytes in rapid and slow disease. AB - Thirteen adults with acute myeloblastic leukaemia (AML) in early 2nd complete remission (CR) were treated with recombinant interleukin-2 (IL-2) and autologous IL-2-activated peripheral blood lymphocytes (LAK cells). All 13 developed IL-2 induced in vitro lymphocytoxicity against K562 and Daudi target cells. After seven years' follow-up, there was no overall improved survival compared with a historical control group treated with chemotherapy alone. However 7/13 patients developed T-cel-associated cutaneous graft-versus-host disease (GVHD), and 4/4 of these tested showed in vitro evidence of a T-cell-mediated graft-versus-leukaemia (GVL) effects. These had significantly longer 2nd CRs and survived longer. More lymphocytes were harvested and more LAK cells were reinfused in these seven cases. Since these patients also had longer 1st CRs, their GVL response to IL 2/LAK cells could be a feature of slowly progressive disease. PMID- 10390075 TI - Upregulation of antitumor immunity by IL-12 gene-transfected AK-5 tumor cells in vivo. AB - We have earlier demonstrated a significant role for IL-12 in the regression of a rat histiocytic tumor, AK-5. In order to analyze further the antitumor immunity induced by interleukin (IL)-12, we have established IL-12-secreting tumor cell clones by gene transfection. Significant enhancement in the lytic potential of splenocytes by the culture supernatants containing IL-12 demonstrated retention of biological activity by the tumor-cell-derived cytokine. Athymic nude mice transplanted subcutaneously with tumor cells engineered to secret IL-12 showed a significant reduction in tumor size, with enhanced antibody-dependent cellular cytotoxicity. Analysis of the serum samples from animals injected with the IL-12 gene-transfected AK-5 cells on different days revealed a significant increase in circulatory IL-12, interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha and antitumor antibodies, all of which contributed to the reduction in tumor mass. The enhanced proliferative capacity of splenocytes from these animals indicated the presence of highly activated immune cells in vivo. Similarly, intraperitoneal transplantation of IL-12 gene-transfected tumor cells in syngeneic Wistar rats induced a significant increase in cellular cytotoxicity, with a concomitant reduction in circulatory IL-12 (p40) protein. Administration of antibodies to IL 12 and IFN-gamma reduced the expression of the costimulatory molecules B7.1 and B7.2 and the cytolytic effectors granzyme B and Fas-L, suggesting their involvement in IFN-gamma-dependent antitumor immune response induced by IL-12. The present study thus demonstrates that IL-12 gene therapy could be among the promising approaches for an effective cancer therapy. PMID- 10390076 TI - Antisense strategy in hematological malignancies. AB - Standard cytotoxic chemotherapy for neoplastic disease is fraught with systemic toxicity. The ratio of the toxic dose to the therapeutic dose is relatively low, which reflects the large number of cellular targets affected by the chemotherapeutic agent as well as its inability to distinguish between normal and malignant cells. The discovery of oncogenes and tumor suppressor genes involved in the process of transformation of normal cells into malignant cells has opened new areas of research in oncology, aimed at discovering drugs that could selectively inhibit their biological effects. This therapeutic modality, called an antisense strategy, has become a powerful tool for selectively reducing the expression of target genes in vitro, and there is increasing interest in the possibility of using the same technology in vivo for therapeutic purposes. In oncohematology, a number of trials have been initiated with antisense oligonucleotides directed against molecular targets, including the bcl-2, c-myc, bcr-abl, c-myb or p53 oncogenes and tumor suppressor genes. The experience gained from these studies will be applicable to the next generation of antisense compounds, which may include oligonucleotides with novel backbones or other structural modifications, as well as for expansion of the use of antisense oligonucleotides in combination approaches for the treatment of hematological malignancies. PMID- 10390077 TI - Biological and clinical implications of interleukin-7 and lymphopoiesis. AB - Interleukin 7 (IL-7) is a stromal cell-derived cytokine that stands out as being the only cytokine identified to date on which development of B and T lymphocytes is absolutely dependent. IL-7 functions primarily as a growth and antiapoptosis factor for B- and T-cell (alphabeta and gammadelta TCR+ cells) precursors, and is essential for differentiation of gammadelta TCR+ cells. IL-7 can function as a cofactor during myelopoiesis, and is capable of activating monocytes/macrophages and natural killer (NK) cells. Its receptor (IL-7R) is a heterodimer of an alpha chain that specifically binds IL-7 and the common gamma chain gammac that is also a component of the receptors for IL-2, IL-4, IL-9 and IL-15. The functions of IL 7 in normal lymphocyte development and activation have led to the demonstration of the ability of IL-7 to stimulate lymphopoiesis in lymphopenic mice, suggesting a possible clinical application of IL-7 in accelerating lymphoid reconstitution in lymphopenic patients. There have also been a number of preclinical studies pointing to the possible utility of IL-7 in antitumor clinical applications, and clinical trials involving IL-7 gene therapy of metastatic disease are underway. IL-7 has also been shown to promote engraftment of stem cells in mice receiving bone marrow transplants, pointing to a possible use of IL-7 in patients receiving bone marrow or peripheral blood stem cell transplants. Areas of IL-7 biology that are essentially unexplored include the mechanisms of regulation of the expression of IL-7 and IL-7Ralpha, as well as the mechanisms by which IL-7 is a growth and differentiation factor for gammadelta T cells but a growth factor only for alphabeta T cells. PMID- 10390078 TI - TNF-alpha in acute cardiac transplant rejection. AB - Acute cardiac allograft rejection is an immune-mediated response, hallmarked by cellular infiltration and myocyte damage in the transplanted heart. Cardiac biopsy sampling has been the 'gold' standard for routinely monitoring episodes of acute rejection. As cardiac biopsy is invasive, attention has focused on other non-invasive methods, such as serum analysis, for monitoring purposes. Tumour necrosis factor alpha (TNF-alpha) is a lymphocyte- and macrophage-derived cytokine that is pleiotropic in its actions. Its proinflammatory functions suggest that it may play an important role in initiating and orchestrating the rejection response. Studies demonstrating a correlation in the expression of TNF alpha with the severity of the rejection episode have placed TNF-alpha as a prime candidate marker of rejection, and have prompted further study to elucidate these findings. This review discusses the limitations of the methodologies used to identify TNF-alpha, and how intragraft expression of TNF-alpha is not reflected in the serum. Furthermore, we describe how other stimuli besides the rejection response can affect TNF-alpha production, arguing against its use as a 'rejection specific' marker. Nevertheless, genetic studies suggest that TNF-alpha may influence transplant outcome, and offer a new tool for studying the role of TNF alpha in acute transplant rejection PMID- 10390080 TI - Statistical methods in epidemiology. III. The odds ratio as an approximation to the relative risk. AB - PURPOSE: This paper introduces readers to case-control studies and how to analyse them. Specifically, the odds ratio statistic is discussed. The importance of the odds ratio in epidemiology is that it is used as an approximation to the true relative risk. METHOD: Data are presented in the form of 2 x 2 contingency tables, and a method for calculating the odds ratio is presented. An extension to 2 x k tables (where k > 2) is given, as is a graphical method for plotting odds ratios. Confidence intervals based on the Normal approximation are introduced. RESULTS: Some properties of the odds ratio statistic are illustrated by taking examples from the author's own teaching experiences. CONCLUSION: As long as the odds ratio is not used uncritically as an estimate of the relative risk, it remains an attractive statistic for epidemiologists to calculate. PMID- 10390079 TI - Cytokines (IFNs, TNF-alpha, IL-2 and IL-12) and animal models of cancer. AB - Cytokines are a complex family of mediators that influence many aspects of tumour cell biology. Whether they are used as a therapy or produced locally during the process of tumorigenesis, they are able to act on tumour cells, on the tumour stroma and on the host itself. Animal models have played key roles in optimizing doses and schedules, and have been useful in determining net effects in the tissue microenvironment. Whilst early studies used xenogeneic and syngeneic systems, more recently gene and protein delivery systems have been used to introduce ectopically expressed high and continuous levels of cytokines. This review will discuss the application of some animal model systems that have been investigated to further understand the role cytokines play in cancer, and will concentrate on those cytokines for which there are the most experimental animal model data. PMID- 10390081 TI - How does a cerebral stroke affect quality of life? Towards an adequate theoretical account. AB - PURPOSE: To contribute, through a hypothesis-generating, qualitative study, to a consistent theoretical account of the mechanisms by which strokes affect the quality of lives of patients. METHOD: A strategic subsample of six persons (65-85 years) was drawn from a larger sample of 60 stroke patients 3 years after stroke. They suffered from some, mostly mild, motor or cognitive impairments, and underwent a semi-structured interview, which was tape-recorded, transcribed and analysed. RESULTS AND CONCLUSION: Only one of the subjects had any familiarity with the QoL concept. When it was rephrased in familiar terms, all but one stated that their QoL had been reduced after the stroke. All the subjects reported considerable bodily changes. The reduced QoL was not, however interpreted as a direct consequence of these, but as a result of the individual's interpretation and evaluation of the changes. It was essential whether the patient compared the present situation to life prior to the stroke, or to a post-stroke reference point. These personal evaluations interacted with the interpretations of the situation by the patients' significant others. PMID- 10390082 TI - Clinical effectiveness of dramatherapy in the recovery from neuro-trauma. AB - PURPOSE: To investigate the clinical effectiveness of a short course of dramatherapy (an eclectic term encompassing all the arts therapies), delivered in a one-to-one interaction, in a sample of 10 patients in a neuro-rehabilitation unit. METHOD: Each participant received five individual one-to-one sessions of therapy over a 5 week period. A semi-structured interview was carried out with each participant following the course. RESULTS: Qualitative analysis of the taped interviews elicited how the therapy contrasted and complemented the rest of the rehabilitation setting and therapies and how it helped psychological adjustment to severe disabilities resulting from neurotrauma. There were four ways in which it appeared to empower the participants and nurture their self esteem. It provided them with a sense of personal space in an otherwise institutional setting; it allowed escapism and enjoyment; it awakened creativity and a sense of potency; and it provided a metaphor to explore personal issues. CONCLUSION: Dramatherapy made an important contribution to the healthy adjustment of some patients both to hospital life and to acquired disability. The reports from the patients indicated that this approach to rehabilitation should be further incorporated and developed in neuro-rehabilitation. PMID- 10390083 TI - The effect of vocational rehabilitation on later sick leave. AB - PURPOSE: The aim of this study was to examine the effect of vocational rehabilitation on later sick leave for employed and unemployed people on long term sick leave. METHOD: The study was based on long-term sick-leave cases initiated during 1992-1994 in Stockholm, Sweden. Of 1704 cases, 383 (321 employed and 62 unemployed) underwent vocational rehabilitation. These were individually matched with sick-leave cases not undergoing rehabilitation, using the variables age, sex, diagnoses and employment status. RESULTS: The hypotheses were (1) that people who underwent rehabilitation, both employed and unemployed, would have less later sick leave than those who did not, and (2) that rehabilitation would affect employed people more than unemployed people. These hypotheses were only partly supported. CONCLUSIONS: The results indicate that vocational rehabilitation has a positive effect regarding later sick leave only for unemployed men. For unemployed women the effect is negative and for those employed, both men and women, rehabilitation has no demonstrable effect. PMID- 10390084 TI - Isolation of speech area syndrome (ISAS): a follow-up study--a rehabilitative approach. AB - PURPOSE: In this study we present our findings concerning the dynamics of language modalities in the isolation of the speech area syndrome (ISAS). The aim of the study was to establish: (1) the linguistic characteristic features of this group of patients; (2) the course of dynamics of language of each patient; (3) whether this course of dynamics is common for the whole group; (4) whether the course of dynamics we found matches with the report in the literature. METHOD: Over a period of 5 years, nine patients with ISAS were diagnosed and treated in our department. They received varying lengths of treatment, and four were followed for 2-6 years after discharge. RESULTS: Despite the common belief that the prognosis for language rehabilitation for these patients is poor, we found that some ISAS patients improved, even dramatically, after prolonged language treatment. CONCLUSION: It is hoped that these findings will alter clinicians' attitudes towards these patients, and provide some hope for rehabilitation outcome. PMID- 10390085 TI - The prevalence of disability from chronic conditions due to injury among adults ages 18-69 years: United States, 1994. AB - PURPOSE: To describe the causes and determine the prevalence of disability from chronic conditions due to injury among US civilian non-institutionalized persons aged 18-69 years. METHODS: Data from the National Health Interview Survey Disability (NHIS-D) Supplement Phase I, United States 1994 were analysed and six disability categories were examined: activities of daily living (ADL), instrumental activities of daily living (IADL), functional activities (FA), sight, hearing, and communication. RESULTS: In 1994, 5.6 million persons aged 18 69 years reported a disability because of a chronic condition that was caused by injury. The prevalence of ADL disability due to chronic conditions caused by injury was 370 per 100000 population; IADL disability was 1256; FA disability was 2512; sight was 231; hearing was 339; and communication was 91 per 100000 population. Fifty per cent of ADL, IADL, and FA disabilities were attributed to motor vehicle crashes and falls, as were 31% of sight, 19% of hearing, and 23% of communication disabilities. CONCLUSIONS: Though these estimates may be conservative, this study indicates that injury is a major cause of disability in addition to a leading cause of death in the US. PMID- 10390086 TI - Increased infant axillary temperatures in non-REM sleep during mother-infant bed sharing. AB - This study addressed the effect of mother-infant bed-sharing on infant body temperature and possible mediating mechanisms. Axillary temperatures were recorded for the entire night in 26 infants on both a bed-sharing night and a solitary sleeping night, accompanied by polysomnography and video-taping to allow assignment of sleep stages and behavioral analysis. All infants were approximately 3 months old, healthy, Latino and breast-feeding; 16 of the infants bed-shared since birth while the others routinely slept alone. Bed-sharing was associated with a significantly increased mean axillary temperature compared to solitary sleeping in both routine bed sharers and routine solitary sleepers. This increase was expressed only in non-REM sleep, with no differences during REM sleep or waking. The increase in temperature during bed-sharing may be related to an increased frequency of transient, movement-associated arousals during bed sharing. PMID- 10390087 TI - Prediction of neurological outcome after birth asphyxia from early continuous two channel electroencephalography. AB - OBJECTIVE: To determine whether two-channel continuous electroencephalography (EEG) applied within 12 h of birth can predict the severity of neurological complications and neurodevelopmental outcome following birth asphyxia. METHODS: A continuous two-channel EEG was performed within 12 h of birth in 22 infants suspected of having suffered birth asphyxia and 11 healthy control infants (22 infants at a general and 11 at a specialist paediatric unit). Criteria to categorise normal and abnormal EEG records were defined and compared with the severity of hypoxic/ischaemic encephalopathy (HIE) and with neurodevelopmental outcome, assessed at or after 12 months of age. RESULTS: EEG recordings were commenced at a median (range) of 2 h 50 min (1 h 45 min to 12 h) after birth. Technically satisfactory recordings were obtained in all but one infant. All control infants remained asymptomatic and had a normal EEG with discernible sleep/awake periods. 12 h after birth the EEG was normal in all 12 infants suspected of asphyxia who remained well or developed grade 1 HIE and was abnormal in six of nine infants with grade II or III HIE. Fifteen of 16 infants suspected of asphyxia with a normal neurodevelopmental outcome had a normal EEG at 12 h; transient abnormalities lasting not more than 8 h had been detected in three of these infants. All five infants who died or developed neurodevelopmental abnormalities had an abnormal EEG. At 12 h of age the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and likelihood ratio for predicting severe (grade II or III) HIE were: 67, 100, 100, 80% and infinity and for subsequent death or neurodevelopmental impairments: 100, 94, 83, 100 and 16%, respectively. Assessment of the EEG before 12 h of age altered prognostic accuracy: 4 h after birth the sensitivity, specificity, positive and negative predictive values and the likelihood ratio for poor neurodevelopmental outcome were 100, 71, 33%, 100 and 3.7%, respectively (16 infants). CONCLUSION: Continuous two-channel EEG is an accurate tool for assessing the severity of neurological insult soon after birth asphyxia. PMID- 10390088 TI - Neuropathological changes in the cerebrum of IUGR rat induced by synthetic thromboxane A2. AB - IUGR was induced by maternal administration of synthetic thromboxane A2 (STA2) from the 13th day of gestation. Fetuses and neonates showed a markedly significant weight reduction. In E16 IUGR brain, no pathological abnormalities were found, but morphological changes appeared in the cortical plate of E18 IUGR brain. In E20 IUGR brain, ectopic clusters of differentiating cells cytologically mimicking neuroblasts were found in the neuroepithelial layer, but these abnormal clusters of cells in IUGR brain of late gestation were never observed in PN7. Morphometric analysis of coronal-sectional areas of the brain and cortical plate demonstrated that there were no differences between IUGR rats and controls in E16 and E18. These areas were, however, significantly reduced in E20 and PN7 growth retarded rats compared with the control. Because the period of STA2 administration coincides with the neuro-developmental stage of cell migration and differentiation, reduction of the uteroplacental blood supply might cause a transient abnormal cytoarchitecture of the cerebral cortex resulting in brain growth retardation. PMID- 10390089 TI - Smoking in pregnancy and children's mental and motor development at age 1 and 5 years. AB - We used data from a Scandinavian prospective multicenter study to investigate if smoking in pregnancy may have an adverse effect on the child's mental and motor abilities. Eligible for enrolment were para I and 2 women with a singleton pregnancy, who resided in one of the study areas and could be registered before the 20th gestational week. Women were classified as 'smokers' or 'non-smokers' at study start. At 13 months, 376 children (124 children of smokers) were evaluated with the Bayley Scales of Infant Development. At this age, children of smokers and non-smokers performed equally well. At 5 years, 369 children (132 children of smokers) were tested with the Wechsler Preschool and Primary Scales of Intelligence Revised (WPPSI-R), and 362 children with the Peabody Developmental Motor Scales (PDMS). Children of smokers had an increased risk of getting a WPPSI R score below the median value of the population (OR = 2.1, 95% CI: 1.2-3.3), but the risk was reduced when we adjusted for maternal education (OR = 1.6, 95% CI: 0.9-3.7). Children of smokers had an increased risk of getting a test score below the median population value on the subscale 'balance' from PDMS (OR = 1.8, 95% CI: 1.2-2.8). Thus, we found that smoking in pregnancy was associated with a small, but demonstrable adverse effect on the child's balance at 5 years, whereas the negative effect on cognitive function did not reach statistical significance, when we adjusted for the mother's level of education. PMID- 10390090 TI - International child care practices study: methods and study population. AB - The study set out to document child care practices in as many different countries and cultures as possible with the aim of providing baseline child care data and stimulating new hypotheses to explain persisting differences in sudden infant death syndrome (SIDS) rates between countries. The protocol, piloted in four countries in 1992, was distributed to 80 potential centres in 1995. Data from 19 centres were received. This paper describes the demographic characteristics of the data from the different centres. Comparison showed significant differences for a number of variables including mean age of completion of the study, response rate, mean gestation, mean birth weight, method of delivery and incidence of admission to neonatal intensive care units. High caesarean section rates identified in the Chinese samples (44 and 40%) were unexpected and have important public health implications. This finding warrants further study but may be related to China's one child policy. We consider that international comparison of child care practice is possible using standardised data collection methods that also allow some individual variation according to local circumstances. However, in view of the heterogeneity of the samples, it will be important to avoid over interpreting differences identified and to view any differences within the qualitative context of each individual sample. Provided there is acknowledgement of limitations, such ecological studies have potential to produce useful information especially for hypothesis generation. PMID- 10390091 TI - Near infrared spectroscopy-measured changes in cerebral blood volume and cytochrome aa3 in newborn lambs exposed to hypoxia and hypercapnia, and ischemia: a comparison with changes in brain perfusion and O2 metabolism. AB - OBJECTIVES: Validation of near infrared spectroscopy (NIRS)-measured changes in cerebral blood volume (deltaCBV) and cytochrome aa3 (deltaCytaa3) as estimators of changes in brain perfusion and oxygenation in the newborn lamb during hypoxia and hypercarbia, and additional hypotension. METHODS AND MATERIALS: In 33 newborn lambs brain perfusion assessed by carotid artery blood flow (deltaQcar: ml/min)and cerebral metabolic rate of oxygen (deltaCMRO2: ml O2/min) were related to NIRS-derived deltaCBV (ml/100 g) and deltaCytaa3 (microM) during combined hypoxia and hypercarbia and additional hypotension. Combined hypoxia and hypercapnia was induced by ventilation with 6-8% of O2 and 10% of CO2 during 30 min, and additional hypotension ( < 35 mmHg for 5 min) was induced by careful withdrawal of blood. RESULTS: CBV increased during hypoxia and hypercarbia, decreased during additional hypotension and was related to deltaQcar: (0.009 ml/100 g change per ml/min Qcar: P < 0.0001). Cytaa3 increased during hypoxia and hypercarbia, decreased during subsequent additional hypotension andshowed a reverse relationship with deltaCMRO2 (-1.65 microM change per ml O2/min CMRO2: P <0.0001). Cytaa3 remained above baseline during reperfusion. CONCLUSIONS: deltaCBV estimates changes in brain perfusion, but overestimates brain perfusion during hypotension. The pattern of deltaCytaa3 suggests less oxygen utilisation by brain tissue during hypoxia and subsequent reperfusion. PMID- 10390092 TI - Risk factors for secondary arrest of labor among women >41 weeks' gestation with an unfavorable cervix undergoing membrane sweeping for cervical ripening. AB - OBJECTIVE: To identify the risk factor(s) for secondary arrest of labor among women with an unfavorable cervix at > 41 weeks' gestation. METHODS: Prospectively all gravid women with a Bishop score of < or = 4 and no contraindication to a vaginal delivery were candidates for this study. Univariate analysis followed by logistic regression modeling were used to identify variables with a significant association. RESULTS: Over a 9-month period, 115 women entered into the study. In univariate analysis, variables with a significant association with cesarean delivery: (1) non-Caucasian race (P = 0.007), Bishop score < 7 at hospitalization; P = 0.001, and reason for admission (P = 0.017). Logistic regression analysis yielded OR 4.7 (1.6, 15) non-Caucasian race and 9.5 (3.2, 30.8) Bishop < 7 on admission. CONCLUSION: Pregnancies > 41 weeks with an unfavorable cervix, non-Caucasian race and a failure to achieve a Bishop score of > or = 7 prior to hospitalization were significant risk factors for abdominal delivery. PMID- 10390093 TI - Sonographically diagnosed pelvic hematomas and postcesarean febrile morbidity. AB - OBJECTIVE: To evaluate the incidence of ultrasonographically-diagnosed postcesarean hematomas and correlate their presence with febrile morbidity. METHODS: Prospective study of 111 consecutive patients who had a pelvic ultrasound 4-6 days post-operatively. Ultrasonographic findings were correlated with clinical data. RESULTS: Postoperative fever was diagnosed in 28 (25%) patients. Fifteen (13.5%) women had hematomas; 10 (9%) had bladder-flap and five (4.5%) had subfascial hematomas. Only subfascial hematomas were significantly associated with post-operative fever (P = 0.01). CONCLUSIONS: Postcesarean bladder-flap hematomas are not predictive of post-operative fever. The presence of subfascial hematomas should be specifically sought in the evaluation of a febrile postcesarean patient. PMID- 10390094 TI - Maternal and perinatal mortality and morbidity associated with transverse lie. AB - OBJECTIVE: The aim of this study of transverse lie in labor of patients admitted to Korle Bu Hospital between 1 January 1996 and 30 June 1998, was to identify the methods of delivery, the perinatal and maternal morbidities and mortalities, and to provide recommendations to improve the outcome. METHODS: This was a retrospective study of 152 patients who presented at the labor wards with transverse lie. The data sources of this study were the antenatal records, the labor wards delivery record books, the postnatal records and the admission books at the Neonatal Intensive Care Unit. RESULTS: One hundred and forty-two cases (92.1%) had an emergency cesarean section. The rest had the external version followed by vaginal delivery. There were two maternal deaths resulting from hemorrhage, infections and difficult surgery. There were 25 stillbirths, and 37 of the neonates required hospital admission. CONCLUSION: Transverse lie carries a high rate of complications in labor. Delivery should be carried out without delay in a hospital well-equipped for cesarean delivery and assisted vaginal delivery. The complications could be further reduced by early diagnosis during the antenatal period when associated risk factors, such as placenta previa could be diagnosed with the aid of the ultrasound scan. Elective deliveries could then be undertaken. PMID- 10390095 TI - Ultrasound guided aspiration of endometrioma--a new therapeutic modality to improve reproductive outcome. AB - OBJECTIVE: To evaluate the therapeutic efficacy and reproductive outcome following ultrasound guided aspiration (UGA) of endometrioma in infertile patients. METHOD: This is a prospective non-randomized clinical report of UGA in 22 infertile patients with endometrioma. The aspiration of endometriotic cysts was carried out transvaginally in nine and transabdominally in 13 patients. Following aspiration intranasal buserilin was given to eight and danazol to 14 patients. In the case of recurrence a reaspiration was done. Patients were allowed to conceive following medical therapy. Recurrence of endometrioma and conception rate was recorded and correlated with endometrioma size and volume aspirated. RESULT: A total of 47 aspirations were done. There were no procedure related complications. Reaspiration was required in six patients and one was operated (total recurrence 7/22--31.8%). During a mean follow-up of 20+/-8.4 months nine patients (40.9%) conceived and eight have already delivered at term. The recurrence risk and the conception rate was not affected by the cyst size or volume aspirated. CONCLUSION: UGA of endometrioma can be an effective and safe alternative therapeutic procedure in infertile patients with endometrioma to improve their reproductive outcome. PMID- 10390096 TI - Hysteroscopy and transvaginal ultrasonography in postmenopausal women with uterine bleeding. AB - OBJECTIVE: To evaluate the accuracy of hysteroscopy and transvaginal ultrasonography (TU), based on a histopathological report from endometrial specimens, in diagnosing endometrial pathology in menopausal women with uterine bleeding. METHODS: Four-hundred and nineteen postmenopausal women with uterine bleeding underwent TU, hysteroscopy and endometrial biopsy. Hysteroscopic and sonographic findings have been evaluated on the basis of the final diagnosis established by histologic examination. Sensitivity, specificity and positive predictive value of TU at an endometrial thickness cut-off point of 4 and 8 mm (double layer technique) and of panoramic hysteroscopy have been detected. RESULTS: Normal and abnormal endometrium was found in 222 and in 197 women, respectively. TU showed sensitivity of 95.1%, specificity of 54.8% and positive predictive value of 63.7% at a cut-off limit of 4 mm. With a cut-off limit of 8 mm the corresponding figures were 83.8%, 81.3% and 79.4%. Hysteroscopy demonstrated a sensitivity of 96.5%, specificity of 93.6% and positive predictive value of 92.6%. The combination of the two diagnostic tools showed a 100% sensitivity, 94.8% specificity and 93.3% positive predictive value. CONCLUSIONS: With cut-off limit of 4 mm, TU can be considered the first choice modality of endometrial investigation in women with postmenopausal uterine bleeding to select patients at risk to carry endometrial pathology. Hysteroscopy is a more accurate technique than TU because of better specificity and must be indicated for all patients showing an endometrial strip more than 4 mm. When an endometrial thickness below 4 mm is detected by ultrasound, hysteroscopy may be indicated on clinical background because of the possibility to miss infrequent (2.5% in our series), but relevant endometrial pathologies. Endometrial sampling should follow hysteroscopic view in all cases showing abnormal or suspicious lesions as well as in all cases with irregularly shaped endometrial lining and/or suboptimal endoscopic vision. PMID- 10390097 TI - Estrogen therapy for depression in postmenopausal women. AB - OBJECTIVE: To look at the modification in depressive mood in postmenopausal depressed women after estrogen replacement therapy (ERT). METHOD: Twelve depressed patients divided into two groups of six women each were studied. One group received conjugated equine estrogens (CEE) 0.625 mg/day; the other did not receive any treatment (control group). Mood was assessed in all the subjects at baseline and at 6 months with Hamilton Rating Scale score and considered as depression when it was > 15. Differences between groups were determined by Mann Whitney U-test, and in each group between baseline and 6-month values with Wilcoxon test. RESULTS: The ERT group had a statistically significant decrease in depressive mood (21 vs. 13 points, P < 0.03), while in the control group no significant change was found. Final Hamilton scale scores were significantly lower (P < 0.05) in those under ERT, when compared with those in the control group. CONCLUSION: Depressive mood decreased after 6 months with CEE, so the prescription of ERT can be useful in postmenopausal women with depressive mood. PMID- 10390098 TI - Timing of suicide attempts by self-poisoning during pregnancy and pregnancy outcomes. AB - OBJECTIVE: The purpose of this study was to examine the timing and consequences of suicide attempts by self-poisoning during pregnancy. METHODS: A population based prospective study was organised in the toxicological in-patient hospital in Budapest which is responsible for health services for adult inhabitants poisoned by ingesting chemicals in Budapest and the surrounding area involving 3 million people between 1985 and 1993. All women aged between 16 and 50 years (22969) who were admitted to the study hospital due to suicide attempts by drug ingestion were examined by a sensitive serum pregnancy test. RESULTS: Of 559 self-poisoned pregnant women, two died. The peak period of suicide attempts was found to be in the first postconceptual month and its majority resulted in a very early fetal loss. The second highest figure was recorded in the second postconceptual month. Thus, 61% of suicide attempts occurred before the third postconceptual month. Later pregnancies had a significantly lower proportion of attempting suicide parallel with advanced fetal development. CONCLUSIONS: Most suicide attempts by self-poisoning occurred after the early recognition of unwanted pregnancies and most resulted in a very early fetal loss. Pregnancies with advanced gestation months had a significantly lower proportion of attempting suicide. PMID- 10390099 TI - Laminaria and sulprostone in second trimester pregnancy termination for fetal abnormalities. AB - The efficacy and safety of intracervical placement of laminaria and intravenous prostaglandin E2 (sulprostone) infusion for termination of second-trimester pregnancies with abnormal fetuses was investigated. One hundred and six pregnant women at 13-29 weeks' gestation with fetal anomalies underwent laminaria tent insertion into the cervical canal on admission. The next morning, Sulprostone infusion was started at a rate of 16 microg/h and increased by 16 microg/h every 30 min to induce uterine contractions. Induction-to-abortion time (IAT), success and complete abortion rates, and sulprostone-related side effects were registered. The overall success and complete abortion rates within 24 h were 91.5 and 80.2%, respectively. The mean IAT was 12.1+/-7.6 h. The incidence of nausea and/or vomiting was 17.9%, with 1.7 episodes per case. Diarrhea and fever (9.5%) were not common. Laminaria tent insertion plus sulprostone infusion was an effective and safe regimen for second-trimester termination of pregnancy with live fetuses. PMID- 10390100 TI - Factors affecting the contraceptive choice in a developing country. AB - OBJECTIVES: To evaluate the factors affecting the contraceptive choice of women in a developing country. METHODS: Demographic characteristics, education and income level, previous and current contraceptive choices of the women from a maternity and a university hospital were retrospectively reviewed for 2 years. The data obtained from the two hospitals were analyzed by Student's t- and chi2 tests. RESULTS: Family planning services were offered to 651 and 7427 women in the university and the maternity hospital, respectively. Although the mean ages and income levels of the women in two centers were similar, the women in the university hospital had lower mean gravidity and mean number of living child, while they had higher education level and previous modern contraceptive use (P < 0.05-0.001). The women in the university hospital more frequently preferred combined oral contraceptive and surgical sterilization, while those in the maternity hospital chose condom and intrauterine device (P < 0.01-0.001). CONCLUSIONS: Women with higher education level had a lower number of pregnancies and living children due to more frequent use of previous effective contraception and they chose combined oral contraceptives and irreversible methods more frequently. PMID- 10390101 TI - Jaundice in pregnant Nigerians. PMID- 10390102 TI - A diabetes-in-pregnancy center in Georgia (former USSR). PMID- 10390103 TI - Laparoscopic surgery in the Cameroon. AB - A new surgical section of operative laparoscopy was installed in General Hospital of Yaounde, Cameroon, in April 1992 after many years of collaboration between the University of Clermont-Ferrand and the University of Yaounde. A total of 735 laparoscopic operations were conducted in the first 5 years. Conditions facilitating such a project are discussed. PMID- 10390104 TI - High hCG level is a risk factor for predicting the occurrence of brain and/or liver metastases from gestational choriocarcinoma. PMID- 10390105 TI - Successful pregnancy in a woman with acromegaly treated with somatostatin analog (octreotide) prior to surgical resection. AB - A 27-year-old woman with a GH-secreting pituitary macroadenoma was treated with continuous s.c. infusion of octreotide prior to surgical resection. Subsequently, she was found to be 6 months pregnant. Fetal echographs were normal, the newborn had no malformation, and postnatal development was normal. PMID- 10390106 TI - Adenomyosis presenting as an adnexal mass after laparoscopic myomectomy. PMID- 10390107 TI - Vaginal bromocriptine administration in patients with hyperprolactinemia. PMID- 10390108 TI - Recurrent spontaneous abortion and selenium deficiency. PMID- 10390109 TI - Human rights and abortion laws. AB - Human rights protections have developed to resist governmental intrusion in private life and choices. Abortion laws have evolved in legal practice to protect not fetuses as such but state interests, particularly in prenatal life. National and international tribunals are increasingly called upon to resolve conflicts between state enforcement of continuation of pregnancy against women's wishes and women's reproductive choices. Legal recognition that human life begins at conception does not resolve conflicts between respect due to women's reproductive self-determination and due to prenatal life. Human rights protect healthcare providers' claims to conscientious objection, but not at the cost of women's lives and enduring health. PMID- 10390110 TI - ACOG committee opinion. Anticoagulation with low-molecular-weight heparin during pregnancy. Number 211, November 1998. Committee on Obstetric Practice. American College of Obstetricians and Gynecologists. PMID- 10390111 TI - ACOG committee opinion. Screening for canavan disease. Number 212, November 1998. Committee on Genetics. American College of Obstetricians and Gynecologists. PMID- 10390112 TI - ACOG committee opinion. Ethical considerations in research involving pregnant women. Number 213, November 1998. Committee on Ethics. American College of Obstetricians and Gynecologists. PMID- 10390113 TI - ACOG practice bulletin. Medical management of tubal pregnancy. Number 3, December 1998. Clinical management guidelines for obstetrician-gynecologists. American College of Obstetricians and Gynecologists. AB - Ectopic pregnancy is a major health problem for women of reproductive age and, in the United States, is the leading cause of pregnancy-related death during the first trimester. Diagnosis and treatment of tubal pregnancy before tubal rupture occurs decreases the risk of death. Early detection may make it possible for some patients to receive medical therapy instead of surgery. Methotrexate, a folinic acid antagonist, has been used to treat patients with small unruptured tubal pregnancies. The purpose of this document is to present evidence, including risks and benefits, about methotrexate as an alternative treatment for selected ectopic pregnancies. PMID- 10390114 TI - Aprotinin attenuates platelet accumulation in ischaemia-reperfusion-injured porcine skeletal muscle. AB - This purpose of this study was to evaluate the effect of aprotinin, a serine protease inhibitor, in ischaemia- and reperfusion-injured myocutaneous flaps and skin flaps. Flap survival, microcirculatory platelet accumulation, and regional blood flow were investigated in seventeen pigs which had been subjected to 8 h of ischaemia and 18 h of reperfusion. The pigs were randomly assigned to aprotinin treatment (n = 9) or saline (n = 8). In-vitro studies were performed to investigate the influence of aprotinin on the activated partial thromboplastin time. The survival of skeletal muscle correlated positively with the concentration of aprotinin (P = 0.02) and could not be explained by regional changes in blood flow. Platelet accumulation was decreased in aprotinin-treated muscle (P = 0.04). In-vitro (n = 10), 100 kallikrein inactivator units/ml aprotinin prolonged the activated partial thromboplastin time both in plasma (P = 0.001) and in blood (P = 0.002), suggesting an anticoagulant rather than a procoagulant effect. In conclusion, aprotinin at high concentrations may be beneficial for the survival of skeletal muscle and provides protection from platelet accumulation in the microcirculation of skeletal muscle exposed to ischaemia and reperfusion injury. PMID- 10390115 TI - Fibrinogen, fibrin and its degradation products in drained blood after major orthopaedic surgery. AB - The aim of this study was to evaluate the fibrinogen enzymatic conversion in blood collected postoperatively from a surgical wound. Ten otherwise healthy patients (aged 11-28 years) in need of surgical treatment for thoracic scoliosis were included in the study. Arterial blood preoperatively and at wound closure were compared with samples of drained blood from the wound at closure and from a collection system for autologous transfusion 2.8 +/- 1.1 h later. There was a decrease in the fibrinogen content in arterial blood from 2.17 +/- 0.35 g/l to 1.23 +/- 0.42 g/l, which followed a 40% haemodilution estimated from the blood loss of 1.6 +/- 0.9 l during the operation. Drained blood contained high concentrations of D-dimer (85 +/- 53 mg/l from the wound and 121 +/- 47 mg/l from the collection system), but no clottable fibrinogen. The Western immunoblots all visualized the same patterns; in drained blood there were split-products mainly from cross-linked fibrin, in contrast to arterial blood which contained only normal fibrinogen. This indicates a strong fibrinolysis in the surgical wound after closure, with concentrations of fibrin degradation products that may impair local coagulation, and if infused, might interfere with general haemostasis. PMID- 10390116 TI - Laboratory diagnosis of thrombophilia by endothelial-cell-modulated coagulation. AB - This study investigated whether the addition of endothelial cells to blood or blood plasma is of value in global laboratory diagnostic testing for thrombotic tendency. Plasma from thrombotic patients and healthy individuals was exposed to human umbilical vein endothelial cells (HUVEC), in monolayers or suspensions, and fibrin deposition or clotting time, respectively, was registered. The latter was determined by a novel rheometric procedure that also gave information about coagulum rigidity. Plasma from patients (n = 10) tended to deposit more fibrin on HUVEC monolayers than plasma from healthy individuals (n = 10). When mixed with suspended HUVEC, plasma from patients (n = 14) showed shorter clotting times than plasma from healthy individuals [n = 13; 4.79 +/- 1.02 min (mean +/- SD) compared with 6.80 +/- 1.50 min, P < 0.001]. Coagulum rigidity among patients also differed from that of healthy individuals (P < 0.05). The study showed that the addition of endothelial cells to blood plasma is of value in global laboratory diagnostic testing for thrombotic tendency. PMID- 10390117 TI - Endothelial release of tissue-type plasminogen activator and ischemia-induced vasodilatation are linked in patients with coronary heart disease. AB - Dysfunction in the vascular endothelium disturbs blood flow and predisposes individuals to atherosclerosis. Deteriorated fibrinolysis may further enhance the risk for atherothrombosis. We investigated 14 healthy volunteers and 24 patients with coronary heart disease. Endothelium-dependent (acetylcholine- and ischemia induced) and endothelium-independent (nitroprusside-induced) vasodilatation in the forearm vasculature were studied using strain-gauge plethysmography, and the fibrinolytic system measured as the response of tissue plasminogen activator (t PA) to provocation testing (20 min venous occlusion; VOT). When acetylcholine induced vasodilatation was measured, endothelium-dependent vasodilatation differed between groups: those with coronary heart disease had a median value of 8.5 ml/min per 100 g tissue (25th to 75th percentile 4.8-10.3), compared with 11.6 ml/min per 100 g tissue (7.3-15.5) among healthy volunteers (P = 0.03). However, ischemia-induced vasodilatation showed no difference between the groups [26.8 (22.7-35.0) versus 29.1 (25.6-30.7) ml/min per 100 g tissue, respectively, NS]. Levels of t-PA after VOT also showed no difference between the groups [21.5 (16.5-31.9) versus 20.4 (11.8-31.5) ng/ml, respectively, NS]. Results of ischemia tests and levels of t-PA after VOT correlated only in patients with coronary heart disease (r = 0.5, P = 0.015), and not in healthy volunteers. We observed a positive correlation between endothelium-dependent vasodilatation function and endothelial release of t-PA. This indicates that the same mechanism that results in defective ischemia-induced endothelial relaxation in patients with coronary heart disease may also result in suppressed fibrinolytic capacity, thus making such patients more prone to atherothrombosis. PMID- 10390118 TI - Catheter-directed intra-arterial thrombolysis: size of soluble fibrin(ogen) degradation products studied by electrophoresis and immunoblotting. AB - The aim of this study was to investigate the proteolytic degradation of fibrin(ogen), with special emphasis on the size of soluble fibrin(ogen) derivatives identified before, during and after intra-arterial catheter-directed thrombolysis with alteplase. Arteriography was performed before thrombolysis and after 0.5, 3, 10 and 24 h in six patients with peripheral native artery or bypass occlusions. Samples collected simultaneously intra-arterially from the thrombus and from venous blood were studied by immunoblotting patterns obtained after polyacrylamide and agarose gel electrophoresis. Supernatant from centrifuged material aspirated from the thrombus before thrombolysis contained soluble fibrin(ogen) derivatives with molecular weights of several thousand kDa. Two types of soluble fibrin(ogen)-related material were visualized during treatment: high molecular weight species (500-1000 kDa) displaying an almost continuous spectrum of molecular weights, suggesting gradual proteolytic degradation of cross-linked fibrin into X-oligomeric material; and X, Y, DD, D and E fragments. The amount and distribution of fragments strongly indicated that preferential fibrinolysis had taken place. The finding of a sustained level of fragments in post-thrombolytic plasma might indicate that insoluble fragments embolize peripherally and subsequently lyse. A close association between angiographical and molecular findings during both successful and failing thrombolysis was demonstrated. PMID- 10390119 TI - Serum apolipoprotein H and its relationship to lipids and other apolipoproteins in normal human men and women. AB - Apolipoprotein H (apo H), also known as beta-2-glycoprotein I, has recently become of considerable interest in the field of haemostasis. Because of the possible involvement of apo H in lipid pathways, we wished to test the hypothesis that the serum concentration of apo H was related to serum lipids and other apolipoproteins in normal individuals. One hundred and twenty-three healthy young subjects (67 women and 56 men) aged 22.7 +/- 0.70 years (mean +/- SD) were recruited. Serum level of apo H was higher in male subjects that in age-matched female subjects (167.6 +/- 47.5 mg/l versus 148.8 +/- 30.5 mg/l; P < 0.04). Serum levels of apo H were significantly correlated with serum concentration of cholesterol, but only in female subjects (r = 0.28, P < 0.02). No significant correlations were found between serum levels of apo H and triglycerides, high density lipoprotein cholesterol, apo A1, apo B or lipoprotein (a) in either of the sexes. PMID- 10390121 TI - Is activated protein C resistance following orthotopic liver transplantation a risk factor for venous thrombosis? PMID- 10390120 TI - Tissue factor de-encryption: ionophore treatment induces changes in tissue factor activity by phosphatidylserine-dependent and -independent mechanisms. AB - Coagulation is initiated on tissue-factor-bearing cells when factor VIIa complexes with membrane-bound tissue factor and activates factors X and IX. Cellular tissue factor activity does not correlate with tissue factor antigen; treatment with calcium ionophore rapidly increases tissue factor activity without increasing tissue factor antigen. Our study examined the effect of calcium ionophore A23187 on tissue factor activity of freshly isolated, lipopolysaccharide-stimulated monocytes and non-transformed human dermal fibroblasts. A23187 increased tissue factor activity on monocytes and fibroblasts in a dose-dependent fashion between 0.1 and 50 micromol/l ionophore. This increase in activity was proportional to an increase in intracellular calcium in monocytes. The increase in tissue factor activity was partially attributable to an increase in phosphatidylserine expression, as measured by increased prothrombinase activity (1.1- to 4-fold) on ionophore-treated cells. The phosphatidylserine-binding protein annexin V decreased tissue factor activity on both ionophore-treated and untreated cells, reflecting the role of phosphatidylserine in tissue factor activity. However, even in the presence of saturating concentrations of annexin V, the tissue factor activity of ionophore treated cells was 1.3- to 11.3-fold higher than that of untreated cells, indicating that the increase in tissue factor activity did not result solely from increased expression of phosphatidylserine. A23187 increased tissue-factor dependent activation of factors IX and X 1.4- to 7-fold on both cell types, indicating that ionophore treatment did not alter factor VIIa/tissue factor substrate specificity. We conclude that the mechanism by which calcium ionophore increases tissue factor activity is not unique to monocytoid or transformed cells. Furthermore, the ionophore-induced increase in activity is not solely the result of increased exposure to phosphatidylserine. Finally, tissue factor de encryption by A23187 does not alter factor VIIa/tissue factor substrate specificity. PMID- 10390122 TI - Invertebrate compounds acting on the hemostatic mechanism: new information. PMID- 10390123 TI - Gastrointestinal effects of nonsteroidal anti-inflammatory therapy. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) are widely prescribed for the treatment of many conditions including rheumatoid arthritis, osteoarthritis, gouty arthritis, the joint and muscle discomfort associated with systemic lupus erythematosus, and other musculoskeletal disorders. Yet, their benefits, which are believed to be a result of their ability to inhibit cyclooxygenase-2 (COX-2), are accompanied by considerable toxicity. NSAIDs' untoward effects are attributed to their inhibition of the constitutively expressed enzyme cyclooxygenase-1 (COX 1), with attendant suppression of the synthesis of prostanoids, substances that mediate key homeostatic functions. Side effects include suppression of hemostasis through inhibition of platelet aggregation, adverse effects in patients with heart failure and cirrhosis, and those with certain renal diseases, as well as complicating antihypertensive therapies involving diuretics or beta-adrenoceptor blockade. Perhaps most importantly, NSAIDs disrupt the gastrointestinal mucosal protective and acid-limiting properties of prostaglandins, frequently leading to upper gastrointestinal erosions and ulceration, with possible subsequent hemorrhage and perforation. These complications can be reduced through identification of patients at risk, with circumspect use of NSAIDs, careful functional monitoring, and, in the case of gastrointestinal toxicity, co administration of such agents as misoprostol or omeprazole. However, these strategies introduce complexity into the treatment paradigm. Moreover, side effects and adverse events may be significantly reduced through the use of COX-2 specific inhibitors, new agents that alleviate pain and inflammation without the liability for adverse events caused by COX-1 inhibition. PMID- 10390124 TI - Nephrotoxicity of nonsteroidal anti-inflammatory drugs: physiologic foundations and clinical implications. AB - Although the prevalence of nephrotoxicity in patients treated with nonsteroidal anti-inflammatory drugs (NSAIDs) is relatively low, the extensive use profile of these agents implies that many persons are at risk. At basal states of normal renal function, the role of renal prostaglandin production for maintenance of stable renal hemodynamic function is relatively limited. Nonetheless, in the clinical setting of reduced renal perfusion as seen in various forms of cardio renal disease, dehydration, and the aging kidney, the adequacy of renal prostaglandin production mediated predominantly by cyclooxygenase-1 (COX-1) and, potentially, by COX-2 enzyme activity becomes of major significance in the activation of compensatory renal hemodynamics. Inhibition of renal prostaglandin production by the use of NSAIDs in these circumstances can potentially lead to the emergence of several distinct syndromes of disturbed renal function. These include fluid and electrolyte disorders, acute renal dysfunction, nephrotic syndrome/ interstitial nephritis, and renal papillary necrosis. In addition, by blunting the homeostatic renal effects of prostaglandins, NSAIDs can adversely influence blood pressure control, particularly during the use of angiotensin converting enzyme (ACE) inhibitors, diuretics, and beta blockers. This is a matter of considerable public health concern, in that some 12 million US citizens are concurrently treated with NSAIDs and antihypertensive drugs. Finally, the risk of congestive heart failure is significantly increased when NSAIDs are given to patients receiving diuretic therapy who have cardiovascular risk factors. Physiologic factors, clinical presentations, diagnostic modalities, and clinical management strategies appropriate to these NSAID-induced renal syndromes are described. PMID- 10390125 TI - Effects of nonsteroidal anti-inflammatory therapy on platelets. AB - Aspirin and other nonsteroidal anti-inflammatory drugs (NSAIDs) can produce a mild, systemic hemostatic defect by inhibiting normal platelet function. Aspirin acetylates and permanently inactivates cyclooxygenase (COX), while nonaspirin NSAIDs reversibly block COX; thus, all of these drugs cause platelet dysfunction by inhibiting the formation of thromboxane A2, a platelet-activating and vasoconstricting eicosanoid. However, spontaneous bleeding complications outside the gastrointestinal tract very rarely result from the use of aspirin and other NSAIDs in individuals who are otherwise hemostatically normal. Most types of surgery are not usually associated with clinically significant bleeding in patients taking these drugs, making it typically unnecessary to discontinue them and thus delay surgery for the purpose of restoring normal hemostasis. Exceptions may include operations at sites where optimal hemostasis is critical, surgical manipulation of the genitourinary tract and oral cavity, and possibly cardiac surgery. Factors that increase the risk of bleeding with aspirin and other NSAIDs include coexisting coagulation abnormalities and the simultaneous use of alcohol or anticoagulants. PMID- 10390126 TI - Role and regulation of cyclooxygenase-2 during inflammation. AB - Prostaglandins are formed from arachidonic acid by the action of cyclooxygenase (COX) and subsequent downstream synthetases. Recently, it has been found that there are two closely related forms of COX, which are now known as COX-1 and COX 2. Although both isoforms of this enzyme convert arachidonate to prostaglandins, there are significant differences in their distribution in the body and their roles in health and disease. The basis for these important differences lies in the genes for COX-1 and COX-2 and the regulation of these genes. COX-1, the predominantly constitutive form of the enzyme, is expressed throughout the body and provides certain homeostatic functions, such as maintaining normal gastric mucosa, influencing renal blood flow, and aiding in blood clotting by abetting platelet aggregation. In contrast, COX-2, the inducible form, is expressed in response to inflammatory and other physiologic stimuli and growth factors and is involved in the production of those prostaglandins that mediate pain and support the inflammatory process. All conventional nonsteroidal anti-inflammatory drugs (NSAIDs) nonspecifically inhibit both COX-1 and COX-2 at standard anti inflammatory doses. The beneficial anti-inflammatory and analgesic effects occur through the inhibition of COX-2, but the gastrointestinal toxicities and the mild bleeding diathesis occur as a result of concurrent inhibition of COX-1. It is important that physicians fully understand the pharmacologic basis for the differential actions of NSAIDs when prescribing them for pain and inflammation. This understanding is also important so that physicians can critically evaluate the basis for, and the emerging data on, COX-2-specific inhibitors and their potential role in clinical medicine. Agents that would inhibit COX-2 while sparing COX-1 represent an attractive therapeutic development and could represent a major advance in the treatment of rheumatoid arthritis and osteoarthritis, as well as a diverse array of other conditions. PMID- 10390127 TI - Cyclooxygenase-2 specificity and its clinical implications. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) reduce pain and inflammation by inhibiting the synthesis of prostanoids. However, these drugs inhibit both cyclooxygenase-1 (COX-1), which is essential for the regulation of homeostasis in many tissues, as well as COX-2, which is an important mediator of pain and inflammation. Disruption of COX-1 enzymatic activity by NSAIDs leads to such side effects as interference with platelet functions and gastric ulcers. The recent development of COX-2-specific inhibitors, such as celecoxib, raises the possibility of relieving pain and inflammation with reduced risk of gastrointestinal complications. In Phase II and III studies, celecoxib has demonstrated efficacy in alleviating dental pain and the signs and symptoms of osteoarthritis and rheumatoid arthritis. This COX-2-specific inhibitor was also associated with a markedly lower rate of gastroduodenal injury than is seen typically with NSAIDs. Incidence of most adverse events (including gastrointestinal) and withdrawal rates resulting from adverse events with celecoxib were similar to placebo. Celecoxib appears to be both safe and effective in the treatment of osteoarthritis and rheumatoid arthritis. Its COX-2 specific inhibitory properties thus introduce the possibility of effective relief of arthritic and other types of pain and inflammation with less risk of the mechanism-based toxicities observed with conventional NSAIDs. PMID- 10390128 TI - The clinical potential of cyclooxygenase-2-specific inhibitors. AB - Emerging evidence suggests that cyclooxygenase-2 (COX-2) has diverse physiologic and pathophysiologic functions. It is expressed constitutively in the developing kidney and brain, playing a role in their proper maturation and function. Further, COX-2 expression may be up-regulated at certain sites: in the kidney during sodium restriction; in the microglia of cognitive centers within the hippocampus and cortex in Alzheimer's disease; and in intestinal adenomas and colon tumors. On the basis of COX-2 expression in Alzheimer's disease and colon cancer, COX-2-specific inhibitors may find clinical utility in the prevention or treatment of these conditions. Despite this apparently optimistic outlook for future uses of COX-2 inhibitors, most of the findings supporting this perspective are based on in vitro and in vivo models and must be rigorously corroborated in human studies, some of which are already planned. PMID- 10390129 TI - Hemostatic factors in hypertriglyceridemic men: effects of a fatty meal before and after triglyceride-lowering treatment with etofibrate. AB - The aims of this double-blind study were to examine whether in hypertriglyceridemic men the ingestion of a standardized fatty meal alters hemostasis negatively and whether triglyceride-lowering treatment with etofibrate for 6 weeks alters fasting and postprandial hemostasis positively, thus reversing the potential negative effects of a fatty meal on postprandial hemostasis. To answer these questions, we measured markers of hemostasis immediately before a standardized fatty meal, and 4, 6, 8, and 10 hours after the meal in 21 hypertriglyceridemic men both before and after treatment with etofibrate. We found that the concentration of plasmin alpha2antiplasmin complex markedly increased for at least 10 hours after the fatty meal, but that the activation of factor XII and the concentration of prothrombin activation fragment1+2 decreased after the fatty meal. These results on factor XII contradict reported in vitro data. Triglyceride-lowering treatment with etofibrate in 10 of these men for 6 weeks increased fasting and postprandial protein C and plasminogen and also slightly decreased the activation of fXII; however, it did not reverse the postprandial increase of PAP or change the decrease of prothrombin activation fragment1+2. Our findings indicate that postprandial lipoproteins alter markers of hemostasis positively in an antithrombotic and profibrinolytic direction. In addition, triglyceride-lowering treatment with etofibrate only slightly improves markers of fasting and postprandial hemostasis in an antithrombotic and profibrinolytic direction. PMID- 10390130 TI - Plasma von Willebrand factor and intestinal ischaemia-reperfusion injury in rats. AB - The purpose of this study was to determine whether plasma von Willebrand factor concentrations are correlated with the degree of intestinal ischaemia-reperfusion injury. Forty-six anaesthetised adult Wistar rats were divided into five groups. The sham-operated group (S, n=10) had laparotomy and isolation of the superior mesenteric artery without clamping. Three ischaemia-reperfusion groups (n=10 in each) had clamping of the superior mesenteric artery for 15, 30, and 45 minutes, respectively, and reperfusion for 15 minutes. A control group (C, n=6) had direct puncture of the heart to sample blood. Mean arterial pressure was measured continuously. Blood was collected at the end of the study to measure von Willebrand factor. The small bowel injury was graded histologically. There was a significant systemic hypotension after declamping in all ischaemia-reperfusion groups, which had a high negative correlation with the histological score (R= 0.46, F=10.1, p<0.003, simple linear regression). Plasma von Willebrand factor was significantly elevated in the three ischaemia-reperfusion groups compared with the control group but not significantly different from the sham-operated group (mean von Willebrand factor concentration (SEM): 156 (29), 283 (29), 295 (25), 381 (44), and 366 (40)% in C, S, ischaemia-reperfusion 15, ischaemia reperfusion 30, and ischaemia-reperfusion 45 groups, respectively). The concentration of von Willebrand factor was not correlated to the histological score (R=0.22, F=1.83, p<0.2) or the degree of hypotension after the removal of the clamp (R=-0.22, F=1.8, p<0.2, simple linear regression). This study shows that von Willebrand factor concentration does not correlate with the degree of intestinal ischaemia-reperfusion injury. It is unlikely that von Willebrand factor can be used as a predictor of disease severity. PMID- 10390131 TI - Prostaglandin E1-induced deconsolidation of thrombin-activated platelet aggregates I: ultrastructure-computer image analysis. AB - We have compared, at an ultrastructural-computer image morphometric level, the relaxation induced by Mg-ethylene-bis-oxyethylenenitrilo-tetracetic acid and prostaglandin E1 on a model of a thrombin-activated platelet aggregate. Mg ethylene-bisoxyethylenenitrilo-tetracetic acid produced a small increase of 5.0% of the intercellular space over the control levels, and a decrease of 10.0+/-1.3% of the cross-sectional area of the platelets, with no apparent cytoskeletal alterations. In contrast, the prostaglandin El-treated preparation shows a 360% increase in the intercellular space and a decrease of the average platelet cross sectional area of 30.0+/-2.0% with marked cytoskeletal alterations. We use the term "deconsolidation" to describe this effect. The enlargement of the intercellular space allows the observation of two types of contacts: (1) a type S (segmental) complex, of approximately 200-nm length that maintains a narrow interplatelet gap of 20-30 nm, filled with a dense intercellular material, and (2) a type R (reticular) complex, formed by scant focal regions of the plasma membrane from opposing platelets that are connected through a mesh of fibrillar or granular material contained within a variable-size space. We hypothesize that deconsolidation is caused by fluid loss from the platelets into the intercellular space. As a result, platelet volume decreases and intercellular space increases. PMID- 10390132 TI - The use of frozen-thawed platelet-derived phospholipids as a confirmatory test for the diagnosis of lupus anticoagulants. Comparison with two commercial confirmatory system tests. AB - Lupus anticoagulants (LAs) belong to acquired circulating anticoagulants interfering with phospholipid-dependent coagulation tests. Owing to the remarkable variability among patients, SSC guidelines recommend more than one test to detect and confirm the presence of LAs. However, this is an expensive procedure and greatly raises the work load of the laboratory. A standardised platelet-derived phospholipid preparation was obtained and platelet neutralisation (PNP) procedures with APTT and DRVVT reagents were performed on plasmas from 16 patients with LAs and from 41 control subjects. In comparisons, STAclot-PNP and DVVconfirm clotting assays were conducted. PNP by using APTT or DRVVT reagents showed intra-assay coefficient of variation of 1.6 and 1.8%, whereas inter-assays coefficient of variations were 6.8 and 5.1%, respectively. A complete concordance was achieved between STAclot-PNP and PNP by using APTT reagents and between DVV-confirm and PNP with DRVVT reagents, demonstrating a high reliability of both the PNP-based assays. This consistency indicates that the standardised platelet-derived phospholipid preparation obtained allows for reliable, reproducible, and cheap PNP-based confirmatory assays for LAs testing. PMID- 10390133 TI - Tissue-type plasminogen activator antigen and consumption of dairy products. The DESIR study. Data from an Epidemiological Study on Insulin Resistance Syndrome. AB - We investigated whether tissue-type plasminogen activator antigen (t-PA-Ag) was associated with intake of meat, fish, or dairy products. The study population comprised 295 women and 299 men aged 30-64 years, which was a random sample from the D.E.S.I.R. (Data from an Epidemiological Study on the Insulin Resistance syndrome) study comprising 5214 men and women in total. T-PA-Ag was measured in fasting blood samples and the habitual intake of foods was assessed by several questions on a food frequency questionnaire. Cross-sectional data were analyzed. The mean t-PA-Ag concentration was 3.28 ng/mL (SD, 1.26) in men and 2.52 ng/mL (SD, 1.22) in women. The concentration of t-PA-Ag was inversely associated with the consumption of milk and milk products in women (p for trend: 0.15) and in men (p for trend: 0.04). The difference between subjects with a low and a high milk consumption was 13% in women and 19% in men. Similar results were observed for consumption of cheese. The concentration of t-PA-Ag was 21 and 8% lower for women and men with a high cheese consumption, respectively, compared to those with a low consumption. Further analyses showed that the association of t-PA-Ag with milk and milk product consumption was independent of cheese consumption and vice versa. No association between meat or fish intake and t-PA-Ag was observed. The results of this study indicate that, if confirmed by others, a high intake of dairy products may influence fibrinolysis by an effect on t-PA-Ag. PMID- 10390134 TI - Facilitation of plasminogen activation by denatured prothrombin. PMID- 10390135 TI - Intestinal ischemia-reperfusion leads to platelet dysfunction. PMID- 10390136 TI - Overexpression of tissue factor pathway inhibitor in aortic smooth muscle cells inhibits cell migration induced by tissue factor/factor VIIa complex. PMID- 10390137 TI - A system for computing neuromorphometry from magnetic resonance images. AB - A new technique for the automated measurement of a variety of gross neuroanatomical structures (such as cerebral ventricles) is described. The system exploits the local statistical properties of dual-echo magnetic resonance images of the brain to produce a fully unsupervised classification of these images into a variety of tissue types. Following this, a simple region agglomeration procedure aggregates the segmented regions by class, and submits these regions to a computational-geometric analysis from which a wide variety of measures are derived. PMID- 10390138 TI - Detection of bursts of microneurographic activity and estimation of burst parameters. AB - Microneurograms are used to study the neural control of the autonomic nervous system. We developed a technique for automatically detecting the times of occurrence of bursts of microneurographic activity and their durations. It involves cross-correlating a rectified and smoothed microneurogram with a triangular shaped waveform as the signal template. Parameters of the derivative of this cross-correlation indicate the times of occurrence and the durations of the bursts of activity. The accuracy of the technique was assessed by comparing its results with those of four experienced investigators and showed significant agreement between the automated technique and the investigators. PMID- 10390139 TI - Multiresolution restoration of medical signals using the renormalization group and the super-coupling transforms. AB - This paper presents a multiresolution approach to the restoration of Magnetoencephalographic (MEG) signals corrupted by colored Gaussian noise. We compare two methods, namely the renormalization group transform (RGT) and the super-coupling transform (ST). We conclude that although the RGT approach requires fewer site updates than the ST approach in order to converge, the ST approach is overall much faster. The multiresolution algorithm was tested with real and simulated data. In the case of simulated data, where the original signal's peak-to-peak value is known, the algorithm worked well with noise levels up to 80% of this value. PMID- 10390140 TI - Nonlinear multivariable modeling and analysis of sleep apnea time series. AB - This paper investigates the modeling and analysis of physiological data recorded from a 49-year-old male and are composed of three time series: blood oxygen saturation, heart rate and respiration. In particular, it is desired to verify if the models estimated from data can distinguish between the dynamics underlying two different breathing patterns (normal breathing and apnea). The estimated models are nonlinear autoregressive, moving average with exogenous inputs (NARMAX) and the regressors used to compose such models are carefully chosen, among hundreds of candidates, by an automatic procedure. The results discussed in this paper suggest that the dynamics underlying the data are nonlinear and basically deterministic. Using estimated models it seems to be possible to quantify the stability of the fixed point in phase space reconstructed using the blood oxygen time series. This, as discussed, could be the basis of an algorithmic monitoring system. PMID- 10390142 TI - Delay in administration of CDDP until completion of AGM-1470 treatment enhances antimetastatic and antitumor effects. AB - The efficacy of cis-diammine dichloroplatinum (CDDP) therapy in combination with continuous administration of angiogenesis inhibitor o-(chloroacetyl-carbamoyl) fumagillol (AGM-1470) was evaluated experimentally using a transplantable rat osteosarcoma line previously established in our laboratory. AGM-1470 (2.5 mg/kg body weight/week) was administered by Alzet osmotic pumps for 2 weeks starting from 7 days after tumor inplantation and CDDP (1.25 mg/kg) was given on days 21 and 24. The number of lung metastatic nodules was counted and the wet weights of the primary tumors were measured 5 weeks after tumor inplantation. Values with administration of CDDP 3 days after discontinuation of AGM-1470 were significantly lower than when the two agents were coadministered (P < 0.05). This animal model should facilitate optimization of the timing of combination therapy. PMID- 10390143 TI - Inhibitory effect of Lovastatin on spontaneous metastases derived from a rat lymphoma. AB - The HMGCoA reductase inhibitor Lovastatin (LOV) has previously shown to abrogate p21ras farnesylation, which is associated with invasive and metastatic abilities in many tumor models. Considering the scarcity of therapeutic resources against metastasis, our objective was to study LOV as an antimetastatic agent on L-TACB rat lymphoma, which as a syngeneic tumor model resembles more closely the situation in human cancer. We also aimed to analyze the effect of LOV on chemoinvasion, motility, metalloproteases secretion, angiogenic capacity, and adhesion to the reconstituted basement membrane Matrigel. Our results showed that LOV caused no effect on cell motility, metalloprotease secretion and neovascularization. Conversely, LOV produced a significant inhibition of invasiveness, which could be a consequence of an impaired cell adhesion to the basement membrane observed. These effects could explain, at least in part, the inhibitory action of LOV on L-TACB rat lymphoma metastases. PMID- 10390144 TI - Transforming growth factor beta1 acts as an inducer of matrix metalloproteinase expression and activity in human bone-metastasizing cancer cells. AB - Bone metastases are a common complication in prostate and breast cancer patients. It leads to extensive morbidity and eventually mortality. Matrix metalloproteinases (MMPs) are known to be involved in the metastatic process. MMP activity can be down-regulated by transforming growth factor beta1 (TGF-beta1), a growth-modulating factor, found in high concentrations in the bone. TGF-beta1 acts through the TGF-beta1 inhibitory element (TIE) element, a cis-acting element found in the promoter region of most MMP genes, with the exception of MMP-2. We used three human cell lines relevant for bone metastases, namely prostate adenocarcinoma PC-3, breast adenocarcinoma MDA-MB-231, and adenocarcinoma cells of unknown origin, Hs696, and one human osteosarcoma cell line, SAOS-2, and showed that in these cell lines TGF-beta1 partially lost its repressing action on MMP expression. TGF-beta1 was able to induce MMP-9 activity and protein expression in all three bone-metastatic tumour cell types, whereas MMP-9 protein levels were repressed in SAOS-2 cells. In PC-3 cells, TGF-beta1 repressed MMP-1 expression, whereas in MDA-MB-231 and SAOS-2 cells, an increase in the expression of MMP-1 protein was detected. Additionally, an increase in MMP-3 expression was observed in Hs696 cells. Expression and activity of the tissue inhibitors of matrix metalloproteinases, TIMP-1 and TIMP-2, were found increased in both PC-3 and MDA-MB-231 cells. With respect to cell proliferation, TGF-beta1 was able to induce a dose-dependent growth inhibition of up to 50% in primary human mammary epithelial cells. However, in none of the tumour cell lines was TGF-beta1 able to suppress growth substantially. Data presented in this paper support the hypothesis that TGF-beta1 can potentially disrupt the balance existing between osteoclast- and osteoblast-derived MMP activity by inducing altered expression of matrix metalloproteinases and their tissue inhibitors derived from bone metastasizing cancer cells. This could eventually lead to skeletal destruction in patients with advanced metastatic disease. PMID- 10390145 TI - Inhibitory effects of bovine lactoferrin on colon carcinoma 26 lung metastasis in mice. AB - In order to determine the effects of the multifunctional iron-binding glycoprotein, lactoferrin (LF), and related compounds on tumor growth and metastasis, bovine LF (bLF), and bLF hydrolysate and lactoferricin (bLFcin), active products generated by acid-pepsin hydrolysis were administered orally to BALB/c mice bearing subcutaneous (s.c.) implants of the highly metastatic colon carcinoma 26 (Co 26Lu). bLF and the bLF hydrolysate demonstrated significant inhibition of lung metastatic colony formation from s.c. implanted tumors without appreciable effects on tumor growth. bLFcin displayed a tendency for inhibition of lung metastasis. On the other hand, bLF did not exert marked anti-metastatic activity in athymic nude mice bearing Co 26Lu, though bLF had a tendency to inhibit the lung metastatic colony formation associated with anti-asialoGM1 antibody (Ab) treatment. AsialoGM1+ and CD8+ cells in white blood cells were increased after treatment with bLF. In vitro, the viability of Co 26Lu-F55 cells was markedly decreased when co-cultured with white blood cells from mice administrated bLF p.o., but recovered on treatment with anti-asialoGM1 Ab or anti CD8 mAb and complement. The results suggest bLF and related compounds might find application as tools in the control of metastasis and that asialoGM1+ and CD8+ cells in the blood are important for their inhibitory effects. PMID- 10390141 TI - Mechanisms of tumor angiogenesis and therapeutic implications: angiogenesis inhibitors. AB - Angiogenesis is the development of new blood vessels from the existing vascular bed. In normal conditions this tightly regulated process occurs only during embryonic development, the female reproductive cycle and wound repair. In contrast, in pathological conditions such as malignant growth, atherosclerosis and diabetic retinopathy, angiogenesis becomes persistent due to an imbalance in the interplay between the positive and negative regulatory signals controlling the process. Thus, the control of tumor neovascularization may lead to new therapeutic approaches. Indeed, several anti-angiogenic drugs are currently undergoing preclinical characterization and/or clinical investigation. Recent achievement has clarified the mechanisms of action leading to pathological angiogenesis and has highlighted the role of hypoxia, growth factors, growth factor-receptors, enzymes and cell adhesion molecules involved in the process. This knowledge has permitted the design of receptor antagonists, adhesion molecule blockers and new targeted vascular approaches including gene therapy. PMID- 10390146 TI - An ultra-metastatic model of human colon cancer in nude mice. AB - An ultra-high metastatic model of human colon cancer was developed in order to represent highly malignant patient disease for which there is no current model. Surgical orthotopic implantation (SOI) of a histologically intact liver metastasis fragment derived from a surgical specimen of a patient with metastatic colon cancer was initially implanted in the colon, liver and subcutaneously in nude mice. This tumor did not metastasize for the first 10 passages. At the eleventh passage, the tumor exhibited metastasis from the colon to the liver, spleen, and lymph nodes. At this time, two selective passages were carried out by transplanting resulting liver metastases in the nude mice to the colon of additional nude mice. After these two passages, the tumor became stably ultra metastatic and was termed AC3488UM. One-hundred percent of mice transplanted with AC3488UM with SOI to the colon exhibited local growth, regional invasion, and spontaneous metastasis to the liver, lymph nodes, and spleen. While the maximum size of the primary tumor was 0.9 g, the metastatic liver was over 9 times the weight of the normal liver with the maximum weight of the metastatic liver over 12 g. Liver metastases were detected by the tenth day after transplantation in all animals. Half the animals died of metastatic tumor 25 days after transplantation. Histological characteristics of AC3488UM tumor were poorly differentiated adenocarcinoma of colon. Mutant p53 is expressed heterogeneously in the primary tumor and more homogeneously in the liver metastasis suggesting a possible role of p53 in the liver metastasis. The human origin of AC3488UM was confirmed by positive fluorescence staining for in situ hybridization of human DNA. The AC3488 human colon-tumor model with its ultra-high metastatic capability in each transplanted animal, short latency and a short median survival period is different from any known human colon cancer model and will be an important tool for the study of and development of new therapy for highly metastatic human colon cancer. PMID- 10390147 TI - Video microscopy of tumor metastasis: using the green fluorescent protein (GFP) gene as a cancer-cell-labeling system. AB - Video microscopy allows dynamic observation of cancer-cell activity in the microcirculation of live animals. However, observation of cancer invasion and metastasis in situ has never been successful because of the lack of a technique for labeling cancer cells. We report here our success with video microscopy of cancer-cell activity, detection of remote metastases and cancer cells endogenously generated using the green fluorescent protein (GFP) gene as a cell labeling system in live animals. As a cell-labeling system, GFP is stable, efficient, and nontoxic, and it is passed on to subsequent generations of cells. This pilot experiment has demonstrated the feasibility of direct observation and documentation of tumor growth and tumor invasion as well as the metastatic activities of cancer cells in live animals. PMID- 10390148 TI - Evaluation of seprase activity. AB - Seprase is a serine protease that is integral to the plasma membrane and is overexpressed by invasive tumor cells (Pineiro-Sanchez et al., J Biol Chem 1997; 272: 7595-601; Monsky et al., Cancer Res 1994; 54: 5702-10). Seprase activity is most often assessed by zymography, which is not a quantitative assay. This study establishes a relatively simple and quantitative method for determining seprase activity. The degradation of a 3H-gelatin substrate is measured in the presence of 5 mM EDTA which inhibits matrix metalloproteinases but not seprase. The quantitative character of the assay was demonstrated using partially purified seprase from chicken embryos, a preparation that lacks detectable matrix metalloproteinase activity. In this assay, release of 3H-gelatin fragments is linear over time for 1.5 microg/assay seprase concentration as well as for preparations concentrated or diluted by five fold (7.5 microg/assay and 0.3 microg/assay respectively). Additional experiments were performed to validate the quantification of seprase activity using the radiographic assay by comparing the results to zymography. Exposure to 22 or 37 degrees C results in maximal seprase activity while exposure to 80 or 100 degrees C completely abolishes seprase activity in both zymography and the radiographic assay. Exposure to 60 degrees C abolished seprase activity as judged by zymography, but about 50% gelatinase activity was observed using the 3H-gelatin substrate. Immunopreciptiation with seprase-specific antibody specifically removed seprase and lowered the seprase activity remaining in the extracts as judged by both assays. Investigation of the seprase that was partially purified from human breast cancer tissue revealed that its specific activity (cpm gelatin fragments released/{mg protein x h}) is five times greater than that of seprase purified from chicken embryos. This assay will be useful for determining the seprase activity in extracts of tumor tissues and cells as well as for identifying inhibitors of seprase. PMID- 10390149 TI - Immunotherapy of mice with an irradiated melanoma vaccine coupled with interleukin-12. AB - Interleukin (IL)-12 can activate cytotoxic lymphocytes, stimulate natural killer cell activity, induce the production of INF-gamma and inhibit the development of various experimental tumors. We previously demonstrated that immunotherapy of melanoma bearing mice with an irradiated melanoma vaccine (IMV) coupled with IL-2 or GM-CSF had beneficial effects against primary melanoma growth and against subsequent spontaneous metastasis. We also had found that treatment of melanoma bearing mice with IL-12 (300 ng/day) for 4 weeks inhibited the development of primary melanoma tumors in 40% of mice. The purpose of this study was to investigate the efficacy of combined therapy of experimental melanoma with an IMV prepared from B16F10 melanoma cells coupled with IL-12 treatment. C57BL/6 mice were challenged subcutaneously in the tail with B16F10 melanoma cells and by the 45th day, more than 50% of the mice had developed visible primary melanoma tumors at the injection site. Subsequent immunotherapy of mice with IMV, when coupled with IL-12, provided partial inhibition of primary melanoma tumor growth. Optimal results against primary tumor growth were observed when IMV therapy was coupled with IL-12 at a dose of 50 ng/day. Combination of IMV with IL-12 at a dose of 100 ng/day significantly reduced melanoma metastasis to the lungs compared with control mice, and an improvement in mean survival time was observed in mice treated with a combination of IMV with IL-12 (300 ng/day). PMID- 10390150 TI - CD44 expression and hyaluronic acid binding of malignant glioma cells. AB - The mechanisms leading to rapid invasive growth of malignant gliomas are poorly understood. Expression of the hyaluronic acid (HA) receptor CD44 and adhesion to HA are involved in invasive properties. Our previous studies have shown that malignant glioma cells are able to adhere to extracellular HA. Here we investigated expression of the hyaluronic acid receptor CD44 protein in five human (T98G, A172, U87MG, 86HG39, 85HG66) and two rat (C6, 9L) glioma cell lines. Influence of anti-CD44 antibody and hyaluronidase-preincubation on the HA-binding was determined using HA/BSA (bovine serum albumin)-coated culture plates. While all gliomas were highly positive for CD44 with no differences in the number of positive staining cells, median fluorescence intensity decreased as follows: C6>T98G>9L>85HG66> 86HG39>A172>U87MG. Using HA/BSA coated culture plates the relative levels of specific adhesion to HA were determined as T98G>A172>9L>86HG39>U87MG> 85HG66. C6 cells failed to bind HA specifically. Incubation with anti-human-CD44 MAb significantly decreased HA-adhesion of T98G, A172, 85HG66 and U87MG human glioma cells. However the binding capacity was completely blocked only in 85HG66 cells. The three other cell lines kept a specific HA-adhesion after saturation of the receptor. Hyaluronidase pretreatment markedly enhanced HA-adhesion of C6 and 9L rat glioma cells. These results suggest that (i) HA-adhesion of malignant glioma cells is mainly, but not only, mediated by CD44, (ii) expression of CD44 does not correspond with adhesion capacity and (iii) cell-bound glycosaminoglycans may influence glioma cell adhesion to extracellular HA. PMID- 10390151 TI - Invasion by esophageal cancer cells: functional contribution of the urokinase plasminogen activation system, and inhibition by antisense oligonucleotides to urokinase or urokinase receptor. AB - Early metastasis contributes to the very poor prognosis of esophageal carcinoma. The recent immunohistochemical finding that invasive esophageal carcinomas express elevated levels of urokinase (uPA) and urokinase receptor (uPA-R) in vivo suggest that the plasminogen activation system may contribute to metastasis in esophageal cancer. The aim of our study was to functionally investigate, at the molecular level, the relative contribution of uPA and uPA-R to the invasiveness of esophageal cancer cells in vitro. The three esophageal cancer cell lines, OC1 3, generated in our laboratory, were analyzed for uPA and uPA-R expression by RT PCR, immunoenzymatic staining, and quantitative ELISA. Invasiveness of all cell lines was quantified as percentage cellular invasiveness in a standardized Matrigel in vitro assay. OC1 and OC3, which were found to coexpress both uPA and uPA-R, displayed stronger invasiveness (44% and 32.5% respectively) relative to OC2 (19%) which expressed uPA-R but was negative for uPA. Transfection of OC2 cells with the uPA cDNA resulted in two variants, OC2.uPA1 and OC2.uPA2, stably expressing functional uPA. Both transfectants exhibited enhanced invasiveness (60% and 50% respectively) relative to the parent uPA-negative OC2 cells (19%). Antisense oligonucleotide inhibition of either uPA or uPA-R expression resulted in a similar, marked reduction in invasiveness of esophageal tumor cells which normally coexpress both molecules (OC1, OC3 and the uPA-expressing OC2 transfectant clones). Neither antisense treatment altered the basal invasiveness of OC2, which expresses uPA-R but not uPA. In conclusion, coexpression of uPA with its receptor, uPA-R, is required for functional involvement of the urokinase system in invasion by esophageal carcinoma cells. Our results suggest that these synergistic mediators of invasiveness are quantitatively major contributors to the invasiveness of esophageal carcinoma. PMID- 10390152 TI - Drosophila center divider gene is expressed in CNS midline cells and encodes a developmentally regulated protein kinase orthologous to human TESK1. AB - The Drosophila center divider gene (cdi) was isolated in an enhancer trap screen undertaken to identify genes involved in embryonic central nervous system (CNS) midline cell development. Three independent lines with P-element insertions at 91F were analyzed that all showed prominent beta-galactosidase expression in the CNS midline precursor cells and other cell types. Null mutations were created by imprecise P-element excision and shown to be larval lethal, although no severe CNS defects were observed in mutant embryos. The DNA surrounding the sites of insertion was cloned and found to contain a transcription unit that was dynamically expressed in a pattern corresponding to the enhancer trap line beta galactosidase expression. Sequencing of cDNA clones revealed that the cdi gene encodes a 1140-amino acid protein that is an ortholog of the mammalian testis specific TESK1 protein kinase. This serine/threonine kinase is distinct from other protein kinases because of sequence differences in the residues conferring substrate specificity. The unique sequence is conserved in Cdi, suggesting that Cdi/TESK1 represents a novel class of signaling proteins. PMID- 10390153 TI - Expression of alpha- and beta-tubulin genes during growth of Trypanosoma cruzi epimastigotes. AB - The expression of tubulin genes was studied during the growth of epimastigotes of Trypanosoma cruzi. Northern blot analysis showed that there was a decrease in the levels of alpha- and beta-tubulin mRNAs as epimastigotes changed from the logarithmic to the stationary phase. The changes were associated with a similar decrease in the rates of transcription for both of these genes as measured by run on assays using permeabilized parasites. In contrast to these results, ubiquitin transcription increased slightly. The levels of alpha-tubulin protein per parasite also decreased in stationary compared with logarithmic phase epimastigotes, in close agreement with the decrease in transcription. However, beta-tubulin protein levels did not change significantly. Our results thus indicated that during the growth of epimastigotes, the expression of alpha tubulin is controlled partially at the transcriptional level. On the other hand, the experiments also suggested that beta-tubulin expression is controlled at a post-transcriptional level. PMID- 10390154 TI - A His2AvDGFP fusion gene complements a lethal His2AvD mutant allele and provides an in vivo marker for Drosophila chromosome behavior. AB - We have generated Drosophila melanogaster lines carrying a modified genomic fragment which encodes the D. melanogaster variant H2A.F/Z class histone, His2AvD, fused to the green fluorescent protein (GFP) of the jellyfish Aequorea victoria. We show here that the fusion protein consists of functional GFP and functional histone His2AvD. The His2AvD portion of the fusion gene was shown to be functional by rescue of His2AvD mutant lethality. Fluorescence of the fusion protein in vivo was observed in embryonic cleavage stage interphase nuclei and on chromosomes as early as cycle 9, correlating with activation of transcription. Unlike transcription factors, the His2AvDGFP protein remained on transcriptionally inactive chromosomes throughout mitosis. Subsequently, fluorescence was observed in nuclei at all stages of embryonic and larval development and in adult somatic tissues, consistent with the distribution of His2AvD observed by immunohistochemical staining. This functional fusion histone acts as an excellent in vivo marker for chromosomes and chromosome behavior and, given the ability of the fusion gene to prevent null-mutant lethality, without disrupting normal cellular functions. The very high level of conservation of the H2A.F/Z family of variant histones suggests that the equivalent fusion protein construct should function equally well in a wide range of organisms. PMID- 10390155 TI - Unique features of Chinese hamster S13 gene relative to its human and Xenopus analogs. AB - We have cloned and sequenced the ribosomal protein S13 gene from the Chinese hamster fibroblast HA-1 cells. The predicted protein encoded by this gene is identical to the human ribosomal protein S13, except for one amino acid substitution at residue 29, which is an alanine in the hamster protein and a threonine in that of humans. The physical organization of the six exons and five introns in the hamster S13 gene is also identical to that found in the human and Xenopus genes with respect to the amino acid codes, even though there are small differences in the lengths of the introns. The striking feature is that unlike its human and Xenopus counterparts, which encode two U14 snoRNAs in two separate introns, the hamster S13 gene encodes no U14 snoRNA. Instead, the hamster gene has a pseudo-U14 coding sequence in its third intron. Our data support the idea that the single copy of the hsc70/U14 gene, which we had previously characterized, is the only source for the production of both U14 snoRNA and hsc70 mRNA species in hamster HA-1 cells. PMID- 10390156 TI - Regulation of the insulin gene by glucose: stimulation of trans-activation potency of human PDX-1 N-terminal domain. AB - The beta cells in pancreatic islets of Langerhans increase insulin gene transcription in response to glucose. The pancreatic and duodenal homeobox-1 (PDX 1) plays a major role in glucose-induced insulin transcription. We studied the functional regions of the human PDX-1 protein fused to the DNA-binding domain of the transcription factor Gal4. The results indicate that the N-terminal domain of the hPDX-1, required for transactivation (amino acids 1-120) in transfected betaTC6 and HeLa cells, is also regulated by extracellular glucose concentrations in transfected rat islets. Deletion analyses have led to the mapping of two regions within the N terminus that are essential for its trans-activation properties. One sequence spans amino acids 97-120 in transfected islet and HeLa cells or amino acids 77-120 in betaTC6 cells; the other includes the highly conserved B box (amino acids 31-41). We thus present evidence of a glucose effect on hPDX-1 trans-activation activity, in addition to the previously described regulatory effect on its DNA-binding activity. PMID- 10390157 TI - Abnormal restriction pattern of PIP gene associated with human primary prostate cancers. AB - The PIP gene, localized in the 7q34 region that contains a number of fragile sites such as FRA 7H and FRA TI, codes for gp17/PIP, a protein secreted by breast apocrine tumors. We analyzed the integrity of this gene in 20 tumors of the urogenital tract. We found rearranged EcoRI fragments in 5 of 15 primary prostate carcinomas. No rearrangement was found in normal prostates derived from five patients undergoing prostatocystectomy during treatment of bladder cancers. By Southern blot hybridization with PIP gene exon-specific probes, the rearrangements were mapped at or near the 3' end of the gene. These abnormalities were found, not only in the neoplastic cells invading the prostatic tissues, but also in seminal vesicles without histologic tumoral features. These data suggest a critical role of the PIP gene or neighboring genes in prostate cancer. PMID- 10390158 TI - Structure and functional analysis of a tilapia (Oreochromis mossambicus) growth hormone gene: activation and repression by pituitary transcription factor Pit-1. AB - A gene encoding the Tilapia mossambica (Oreochromis mossambicus) growth hormone (tiGH) was isolated and sequenced. The gene spans 5.6 kb, including 3.7 kb of 5' and 0.2 kb of 3' flanking sequences and a 1.7-kb transcription unit comprised of six exons and five introns. The gene and the 5' flanking region contain several potential binding sites for Pit-1, a key transcription activator of mammalian GH genes. One of these (-57/-42) is highly conserved in fish GH genes. It activates transcription in pituitary cells and binds Pit-1. Transfection of luciferase reporter plasmids containing either the -3602/+19 tiGH sequence or one of its 5' deletion mutants (-2863/, -1292/, and -463/+19) resulted in strong activity in Pit-1-producing rat pituitary GC cells. A dose-dependent activation of the tiGH promoter was achieved in nonpituitary fish EPC and monkey COS cells cotransfected with a rat Pit-1 expression vector, demonstrating the crucial role played by Pit 1 as an activator of the tiGH gene. Fusion of the tiGH promoter with the beta galactosidase gene led to transient expression specifically in the nervous system of microinjected zebrafish embryos. The activity of the tiGH promoter in GC and EPC cells was strongly repressed by extending its 3' end from +19 to +40, a sequence in which a Pit-1-binding site was identified using gel retardation assays. Point mutations of the site that suppressed Pit-1 binding in vitro restored full tiGH promoter activity. Thus, a Pit-1-binding site located in the 5' untranslated region mediates Pit-1-dependent repression of the tiGH gene. PMID- 10390159 TI - Cloning and characterization of rat BAT3 cDNA. AB - HLA-B-associated transcript 3 (BAT3) was originally identified as one of the genes located within human major histocompatibility complex. It encodes a large proline-rich protein with unknown function. In this study, we found that a fragment of the BAT3 gene product interacts with a candidate tumor suppressor, DAN, in the yeast-based two-hybrid system. We cloned the full-length rat BAT3 cDNA from a fibroblast 3Y1 cDNA library. Our sequence analysis has demonstrated that rat BAT3 cDNA is 3617 nucleotides in length and encodes a full-length BAT3 (1098 amino acids) with an estimated molecular mass of 114,801 daltons, which displays an 87.4% identity with human BAT3. The deletion experiment revealed that the N-terminal region (amino acid residues 1-80) of DAN was required for the interaction with BAT3. Green fluorescent protein-tagged BAT3 was largely localized in the cytoplasm of COS cells. Northern hybridization showed that BAT3 mRNA was expressed in all the adult rat tissues examined but predominantly in testis. In addition, the level of BAT3 mRNA expression was more downregulated in some of the transformed cells, including v-mos- and v-Ha-ras-transformed 3Y1 cells, than in the parental cells. PMID- 10390160 TI - Analysis of conserved microsatellite sequences suggests closer relationship between water buffalo Bubalus bubalis and sheep Ovis aries. AB - The distribution and evolutionary pattern of the conserved microsatellite repeat sequences (CA)n, (TGG)6, and (GGAT)4 were studied to determine the divergence time and phylogenetic position of the water buffalo, Bubalus bubalis. The mean allelic frequencies of these repeat loci showed a high level of heterozygosity among the euartiodactyls (buffalo, cattle, sheep, and goat). Genetic distances calculated from the allelic frequencies of these microsatellites were used to position Bubalus bubalis in the phylogenetic tree. The tree topology revealed a closer proximity of the Bubalus bubalis to the Ovis aries (sheep) genome than to other domestic species. The estimated time of divergence of the water buffalo genome relative to cattle, goat, sheep, pig, rabbit, and horse was found to be 21, 0.5, 0.7, 94, 20.3, and 408 million years (Myr), respectively. Although water buffaloes share morphological and biochemical similarities with cattle, our study using the microsatellite sequences places the bubaline species in an entirely new phylogenetic position. Our results also suggest that with respect to these repeat loci, the water buffalo genome shares a common ancestry with sheep and goat after the divergence of subfamily Bovinae (Bos taurus) from the family Bovidae. PMID- 10390161 TI - Major neurosurgical discoveries in the last 20 years. PMID- 10390162 TI - Neuronavigation and surgical neurology: the beginning of a new age or the end of an old age? PMID- 10390163 TI - Cell transplantation in the central nervous system. PMID- 10390164 TI - Genetics of stroke in rats. PMID- 10390165 TI - First gene involved in glioblastoma progression identified. PMID- 10390166 TI - Search for Alzheimer's disease genes yields new candidates. PMID- 10390167 TI - In a high tech age, is clinical judgment a lost art form? PMID- 10390168 TI - Preoperative PET activation for assessment of motor cortex area in precentral chondroma. AB - BACKGROUND: A main problem in the preoperative planning for precentral tumors is the exact assessment of the spatial relationship between the tumor and the functionally relevant brain areas, which may be difficult using only morphologically oriented imaging (CT, MRI). Therefore, we applied motor activation PET and PET/MRI overlay in a patient with a precentral tumor. DESCRIPTION: We report the case of a 21-year-old woman suffering from progressive right-sided headache and intermittent dysesthesia of the left leg. MRI showed a hypointense tumor with inhomogenous contrast enhancement in the right precentral area. For preoperative assessment of the spatial relationship between the tumor and the motor cortex area, the patient underwent two F-18-fluorodeoxyglucose positron emission tomography (PET) scans (1. resting condition and 2. motor activation of the left leg) and subsequent calculation of subtraction images of activation minus rest. Fusion of PET and MRI data (PET/MRI overlay) was performed for bimodal function and morphology presentation. PET revealed an activation pattern behind and below the tumor, indicating that the motor cortex area was shifted to the back. PET findings were confirmed by intraoperative electrophysiology. Cortical stimulation combined with intraoperative neuronavigation localized the motor area of the left foot and leg exactly at the dorsal border, below and lateral to the lesion. After complete resection of the solid tumor, histopathological examination revealed a chondroma. The postoperative course was uneventful, and the patient was discharged without neurological deficits. CONCLUSIONS: This case shows that biomodal imaging (PET/MRI) provides a noninvasive exact assessment of functionally important cortex areas for preoperative planning in patients with cerebral lesions. PMID- 10390169 TI - The surgical anatomy of the perforating branches of the anterior choroidal artery. AB - BACKGROUND: The available information about certain microanatomic features of the AChA perforators is incomplete. Precise knowledge of these vessels is necessary to understand the consequences of their occlusion and to safely operate in their region. METHODS: The AChA perforators were microdissected and examined under the stereoscopic microscope in 10 vascular casts and in 20 hemispheres injected with india ink or radiopaque substance. RESULTS: The perforating branches ranged in number from 2 to 9 (mean, 4.6) and in diameter between 90 microm and 600 microm (mean, 317 microm). The most proximal perforator arose 3.2 mm on average caudal to the AChA origin. The most distal (capsulothalamic) perforator varied in size from 200 microm to 610 microm (mean, 431 microm). One or more of the perforators always originated from the AChA (100%), but some of them also from the uncal (33.3%) or parahippocampal branch (10%) of the AChA, either as individual vessels only (70%) or from common trunks (30%). The perforators gave off the peduncular (20%), optic (23.3%), or uncal side branches (26.7%). CONCLUSIONS: Our findings concerning the origin, position, number, size, branching, penetration site, and relationships of the AChA perforators gave the anatomic basis for safe operations in patients with AChA aneurysms or mediobasal limbic epilepsy. PMID- 10390170 TI - Choroid plexus papilloma: a clinicopathological study of 23 cases. AB - BACKGROUND: Choroid plexus papillomas (CPPs) are rare, accounting for less than 1% of all intracranial tumors in adults. However, they are relatively more common in childhood and constitute 1.5 to 4% of intracranial tumors. DESCRIPTION: They are most often located in the lateral ventricle, followed by the fourth and third ventricles and, rarely, in the cerebellopontine angle. The radiological appearance of a CPP as a cyst with a mural nodule is a curiosity. Bone formation is rare in CPPs and only 6 cases have been described in the literature. Neuromelanin production is also extremely rare and only 2 cases have been reported to date. CONCLUSION: In the present communication, 23 cases of CPP are analyzed and rare clinical, pathological, and radiological features are described. PMID- 10390171 TI - Gamma knife surgery for skull base meningiomas. The effectiveness of low-dose treatment. AB - BACKGROUND: The surgical removal of skull base meningiomas has a high morbidity rate, even by modern microsurgical standards. We evaluated the results of gamma knife surgery for skull base meningiomas using a relatively low radiation dose for the tumor margins. METHODS: We reviewed 24 cases of skull base meningiomas during a 30-month period. The locations of the tumors were the petroclival region in 11 cases, the cavernous sinus region in 9 cases, and the cerebellopontine angle region in 4 cases. Eight patients (33%) had been operated on previously and fourteen patients (67%) had been treated by neuroimaging. The marginal doses for the tumors were 8 Gy to 15 Gy (median, 10.6 Gy). A large petroclival tumor 58 mm in diameter was treated with a staged treatment protocol with a 6-month interval between treatments. RESULTS: Tumor regression was observed in 46% of the patients imaged during the follow-up period (median, 17.1 months). No patients revealed tumor growth in the follow-up period (100% tumor control rate). Eleven patients (46%) had improved clinically by the time of the follow-up examinations. Preexisting cranial nerve deficit in one patient worsened because of radiation injury. CONCLUSION: Although a longer follow-up period is required, the relatively low minimum tumor radiation dose treatment for skull base meningiomas using a gamma knife seems to be an effective treatment with low morbidity. PMID- 10390172 TI - Metastatic adenocarcinoma to the brain mimicking hemorrhage: case report. AB - BACKGROUND: Computerized tomography (CT) of metastatic adenocarcinoma to the brain usually shows low-to-moderate attenuation. However, mucinous adenomas may appear with high attenuation, mimicking hemorrhage. CASE DESCRIPTION: A 68-year old man with a history of metastatic esophageal adenocarcinoma presented to the emergency room complaining of a chronic, progressive right occipital headache. A head CT demonstrated a moderate-to-high attenuation, homogenous mass in the right cerebellar hemisphere consistent with an intracerebral hemorrhage. There was no frank calcification in the mass by CT criteria. An emergent posterior fossa craniectomy revealed nonhemorrhagic metastatic mucinous adenocarcinoma. CONCLUSION: Moderate-to-high attenuation, noncalcified brain masses should raise the possibility of mucin-containing neoplasm. PMID- 10390173 TI - Mesenchymal extraskeletal chondrosarcoma of the orbit. Report of a case and review of the literature. AB - BACKGROUND: Extraskeletal mesenchymal chondrosarcoma (MCS) is relatively uncommon. Orbital location is extremely rare: only 16 cases have been reported until now. We report a case of extraskeletal mesenchymal chondrosarcoma in a 27 year-old man and review the literature on its manifestations and management. CASE REPORT: This patient had a 2-year history of progressive proptosis of the right eye. Skull X-ray and CT scan showed intraorbital calcification and a large lesion in the upper right orbit. He was operated three times because of recurrence of the tumor. The last recurrence was observed to have extension to the intracranial region, detected on MRI and CT scan. This secondary extension of the tumor to the intracranial region has not been previously reported. Immunohistochemical analysis for S-100 protein showed focal positivity. CONCLUSION: Mesenchymal chondrosarcoma of the orbit is rare, and secondary extension to the intracranial region has not previously been reported. PMID- 10390174 TI - A rare cause of right atrial mass: thrombus formation and infection complicating a ventriculoatrial shunt for hydrocephalus. AB - BACKGROUND: Thrombus formation around the intracardiac end of the catheter, thromboembolism, and infection are the most important and life-threatening complications of ventriculoatrial shunts. In this article we report a patient with a large right atrial mass that was diagnosed by 2-D echocardiogram and removed via standard median sternotomy and cardiopulmonary bypass. CASE DESCRIPTION: A 63-year-old man who had a right ventriculoatrial shunt was admitted to our department in a septic clinical condition. His hemoglobin was 10.7 grams, white blood cell count was 22,900/mm3, and sedimentation rate was 50 mm/hr. Blood cultures grew coagulase negative staphylococcus. The echocardiogram showed a right atrial mass at the tip of the shunt catheter. The mass had a cystic and "glove-like" appearance and had a pendulous motion in the right atrium. After combined antibiotic therapy for 10 days, symptoms were relieved but echocardiographic findings did not change. A surgical approach was chosen because of the unchanged size of the mass and the risk of pulmonary embolism. First, the distal part of the ventriculoatrial shunt was separated from its pump and a new ventriculoperitoneal shunt was placed. After this, a standard median sternotomy, cardiopulmonary bypass and right atriotomy was performed. The tip of the shunt catheter with the attached pedunculated mass was removed. CONCLUSION: There are few cases of a large right atrial thrombus secondary to a ventriculoatrial shunt in the literature. Because of these serious complications of ventriculoatrial shunting, careful 2-D transthoracic echocardiographic examination should be mandatory for patients with ventriculoatrial shunts. PMID- 10390175 TI - Persistent trigeminal neuralgia after removal of contralateral posterior cranial fossa tumor. Report of two cases. AB - BACKGROUND: Contralateral trigeminal neuralgia as a false localizing sign in patients with posterior cranial fossa tumors is rare. Persistent contralateral trigeminal neuralgia after removal of the posterior fossa expanding lesion with microsurgical exploration of the affected trigeminal nerve root has been described in only a few reports. Displacement of the brainstem and the trigeminal nerve root, arachnoid adhesions, and vascular compression of the nerve root entry zone have been reported as causes of persistent contralateral trigeminal neuralgia. METHODS: One patient developed transformation of the contralateral constant burning facial pain into trigeminal neuralgia after removal of a posterior fossa meningioma. A typical right-sided tic douloureux in our second patient did not disappear after removal of a left acoustic neurinoma. CT scan revealed brainstem displacement to the side of trigeminal neuralgia. Microsurgical exploration in both cases demonstrated the squeezed and distorted trigeminal nerve root and displaced brain stem with no vascular involvement. Both patients underwent partial trigeminal rhizotomy for pain control. RESULTS: Complete disappearance of the trigeminal neuralgia was evident in both cases with postoperative facial sensory loss. The postoperative course in the first case was uneventful; the second patient died from purulent meningoencephalitis. CONCLUSION: Persistent contralateral trigeminal neuralgia after removal of a posterior fossa tumor is caused by distortion of the fifth nerve root by the displaced brainstem. Partial trigeminal rhizotomy can be performed for alleviation of facial neuralgic pain in cases without neurovascular compression. PMID- 10390176 TI - Anesthetic and surgical experience in a case of total vertical craniopagus. AB - BACKGROUND: A case of craniopagus twins is presented. The twins were attached to each other at the parietal vertex with an interaxis angle of 180 degrees and an interface angle of 90 degrees. To assess the bony, vascular, and nervous system interconnections, the twins underwent computerized tomographic scanning and cerebral angiography under inhalational general anesthesia. CASE DESCRIPTION: To provide adequate skin, tissue expanders were implanted under the scalp at 3 and 7 months of age. Separation became necessary at 1 year of age on an emergency basis, because of respiratory complications in the small twin. Profuse bleeding and hypovolemia led to the death of the healthy big twin. While one would expect that at least one of the twins could have been saved, the small twin also succumbed to air emboli and hypotension hours later. Postmortem examination revealed that the brains were joined at both medial and lateral hemispheric surfaces and shared a common circumferential sinus. CONCLUSION: This case was quite different as regards cerebral anatomy compared to those already reported in the literature. PMID- 10390177 TI - Arachnoid membrane closure. Prevention of postoperative cerebrospinal fluid leakage. AB - BACKGROUND: Opening the arachnoid membrane causes leakage of the cerebrospinal fluid (CSF). CSF spillage and collection in the subdural, epidural, and/or subcutaneous space are occasionally seen on a postoperative CT scan. Closure of the arachnoid membrane is considered to be unnecessary because most of the fluid in the extracerebral space is temporary and disappears spontaneously. However, in some cases, this CSF collection results in serious complications such as brain compression, extracerebral hematoma, intractable meningitis, and skin problems. In this study for prevention of CSF leakage, the arachnoid membrane was tightly closed with 10-0 nylon after a small opening was made for cerebrovascular bypass surgery. METHODS: The arachnoid membrane closure was performed in 18 patients (20 sides) after bypass surgery between 1994 and 1995. As a control, 51 patients (57 sides) had conventional bypass surgery without the arachnoid membrane closure. RESULTS: The radiological findings revealed that only one (5%) of the patients had a small fluid collection after arachnoid membrane closure, which was significantly lower than the control (49%). CONCLUSION: Arachnoid membrane closure is a simple technique and is effective for prevention of postoperative extracerebral CSF collection. PMID- 10390178 TI - The posterior temporal artery as the recipient in superficial temporal artery to posterior cerebral artery bypass. Technical note. AB - BACKGROUND: While superficial temporal artery (STA) to superior cerebellar artery (SCA) or STA to posterior cerebral artery (PCA) anastomosis has been used for rostral brain stem ischemia, it is reported not infrequently to be associated with serious complications. Although the inferior temporal artery has been proposed as a possible recipient artery for the STA, its advantage is not yet widely recognized. CASE REPORT: A 42-year-old man presented with repeated loss of vision in the left visual field. Angiography disclosed occlusion in the proximal portion of the P2 segment of the right PCA. The second case was a 68-year-old man experiencing swallowing disturbance; the bilateral vertebral arteries were markedly stenotic. Since hemodynamic insufficiency was considered to be responsible for the patients' symptoms, STA-PCA anastomosis was performed using the posterior temporal artery (PTA) as the recipient. The postoperative courses were uneventful with good patency of the bypass. TECHNIQUE: Through a horizontally extended temporal craniotomy with the base of the temporal bone sufficiently drilled away, the inferior aspect of the temporal lobe was searched for a recipient artery for the STA. The anastomosis was performed with less difficulty and at a shallower level, by 20 mm in one case and by 10 mm in the other, than had we anastomosed it to the P2 segment of the PCA. CONCLUSION: Anastomosis of the STA to the PTA is less complicated than anastomosis of the STA to the main branch of the PCA for the treatment of rostral brain stem ischemia. PMID- 10390179 TI - A new technique for the assessment of the draining area of a cerebral vein. AB - BACKGROUND: At present, it is not exactly clear which vein is allocated for drainage of blood to a particular area of the human brain. Knowledge of these draining areas is very important for the understanding of occlusive venous diseases. A method was developed that offers the possibility to investigate the draining area of a cerebral vein, with the help of an animal model. METHODS: Brains of sacrificed rabbits are removed and are anterogradely perfused with a coloring matter. Then a vein chosen at random is occluded and anterograde perfusion is restarted using another coloring substance. The working hypothesis is that the part of the brain that is solely dependent for its drainage of blood upon the occluded vein (the draining area of the vein) will show a deficit in staining after the second perfusion. RESULTS Using the abovementioned technique, no filling defect was seen if a vein was occluded near its entrance into the sinus (N = 8) or at a single point over the cortex (N = 7). If a longer trajectory (10-14 mm.) was obstructed, a clear staining defect was seen in 13 out of 16 hemispheres; the three remaining cases seemed to be technical failures. CONCLUSION: A new method is described to investigate the draining area of a cerebral vein. Although the validity of the method is proven in an animal model, it seems a good technique for investigation of human brains postmortem. Application of this technique will contribute to the understanding of the pathophysiology of venous diseases and also elucidate the role of the venous anastomotic pathways. PMID- 10390180 TI - Orbitozygomatic approach by transposition of temporalis muscle and one-piece osteotomy. AB - BACKGROUND: A more simplified and easier technique for the orbitozygomatic approach is sought. We have developed a new modification to fully expose the temporal base before using one-piece craniotomy. METHODS: By transposing the temporalis muscle underneath the zygomatic arch before osteotomy, the temporal base and the inferior orbital fissure can be fully exposed. Craniotomy is made in one piece with the frontotemporal and orbitozygomatic bone together by using a high-speed drill. RESULTS AND CONCLUSIONS: Osteotomy was easy and the cosmetic result was satisfactory. This technique also allows better access to the subtemporal region without removing the zygomatic arch. PMID- 10390181 TI - Cavernous angiomas of the brain stem. Intra-axial anatomical pitfalls and surgical strategies. AB - BACKGROUND: We review the surgical anatomy of the brain stem in relation to the surgical approaches adopted for treatment of cavernomas and identify possible "safe entry zones" on the anterior face of the brainstem. METHODS: Twelve symptomatic patients with cavernoma or telangectasia of the brain stem were surgically treated. The brain stem was divided into the following anatomical areas: ventral medulla, dorsal medulla, dorsal pons, ventral pons, ventral mesencephalon, and dorsal mesencephalon, so that the surgical approach could be "individualized" according to the position of the cavernoma, the nerve fasciculi and nuclei. RESULTS: On the anterior surface of the brain stem a medullar paramedian oblique access to the anterolateral sulcus and a paramedian sagittal pons access seem to avoid the main nerve fasciculi and nuclei. CONCLUSIONS: Although the parenchymal window produced by the cavernoma is the most important parameter for the choice of approach, fairly safe entry zones may be identified even on the anterior surface of the medulla and pons. PMID- 10390182 TI - Spinal cord arteriovenous fistulas involving the conus medullaris: presentation, management, and embryologic considerations. AB - BACKGROUND: Spinal cord arteriovenous fistulas (SCAVF) are uncommon congenital lesions that usually involve the most caudal aspects of the cord. We present three cases of SCAVF that illustrate the clinical manifestations and possible management options. The characteristic involvement of the conus medullaris and an associated tethered spinal cord in one of our patient suggests that a disorder of secondary neurulation may be involved in the formation of these arteriovenous shunt lesions. METHODS: Review of records and radiologic studies in three consecutive patients with SCAVF's treated at this institution. RESULTS: All three patients had SCAVF involving the lower lumbar spinal cord segments or the conus. One of the conus lesions was associated with tethering of the spinal cord. One small lesion (Type A) was treated surgically, whereas the two larger lesions (Type B) were treated using interventional neuroradiologic techniques. CONCLUSIONS: Both surgical and endovascular method have a role in management of these unusual spinal cord vascular malformations. The association with tethered cord suggests that the propensity for SCAVM to occur in the most caudal portions of the spinal cord may result from failure of secondary neurulation to properly develop the unique and complex vascular anatomy of the region. PMID- 10390183 TI - Eric Oldberg. PMID- 10390184 TI - Autopsies. PMID- 10390185 TI - Why Medicare is like a government-sponsored furniture store. PMID- 10390186 TI - Regulation of hematopoiesis in a sea of chemokine family members with a plethora of redundant activities. AB - The field of chemokine biology is a rapidly advancing one, with over 50 chemokines identified that mediate their effects through one or more of 16 different chemokine receptors. Chemokines, originally identified as chemotactic cytokines, manifest a number of functions, including modulation of blood cell production at the level of hematopoietic stem/progenitor cells and the directed movement of these early blood cells. This report reviews chemokines and chemokine/receptor activities mainly in the context of hematopoietic cell regulation and the numerous chemokines that manifest suppressive activity on proliferation of stem/progenitor cells. This is contrasted with the specificity of only a few chemokines for the chemotaxis of these early cells. The large number of chemokines with suppressive activity is hypothesized to reflect the different cell, tissue, and organ sites of production of these chemokines and the need to control stem/progenitor cell proliferation in different organ sites throughout the body. PMID- 10390187 TI - Defective expression of the SHP-1 phosphatase in polycythemia vera. AB - The SHP-1 phosphatase associates with the receptors for erythropoietin, stem cell factor, and interleukin-3, and negatively regulates the mitogenic signals generated during engagement by their respective ligands. The erythroid progenitors of patients with polycythemia vera are hypersensitive to the mitogenic effects of these growth factors despite the fact that the numbers and binding affinities for their receptors are not increased. To determine whether post-receptor signaling defects may account for growth factor-hypersensitivity in polycythemia vera, we determined the expression of SHP-1 in highly purified erythroid progenitors from polycythemia vera patients. Our data demonstrate that in approximately 60% of the patients, expression of SHP-1 in the colony forming unit-erythroid population is diminished. The decreased expression of the protein may result from a transcriptional defect, as suggested by the diminished SHP-1 mRNA expression in the erythroid progenitors of these patients. Studies to determine the level of maturation of polycythemia vera and normal cells indicated that there was no difference between the two at early colony forming unit erythroid stage of differentiation although polycythemia vera cells showed retarded differentiation kinetics at late colony forming unit-erythroid stage of differentiation. Furthermore, SHP-1 expression in normal colony forming unit erythroid demonstrated downregulation of mRNA and protein levels during terminal differentiation, suggesting that its function is required for growth control during the early stages of erythropoiesis. These results indicate an important role for SHP-1 in the regulation of normal human erythroid progenitors and suggest that defective expression of the protein may contribute to the pathogenesis of polycythemia vera. PMID- 10390188 TI - Inhibition of human erythroid colony formation by ceramide. AB - In previous studies, we have demonstrated that the inhibitory effects of tumor necrosis factor (TNF) and interleukin (IL)-1 on human erythroid colony formation are indirect and mediated by beta and gamma interferon (IFN), respectively, which act directly upon erythroid colony forming units (CFU-E). The in vitro inhibitory effect of gammaIFN but not betaIFN is reversed by exposure to high concentrations of recombinant human (rh) erythropoietin (EPO). Ceramide, a product of sphingomyelin hydrolysis, is a known mediator of apoptotic effects of TNF, IL-1, and gammaIFN. In this report, the effects of ceramide on CFU-E colony formation and its implication in the model described above are evaluated. Endogenous ceramide produced by exposure to bacterial sphingomyelinase (0.2-2.0 U/mL) and exogenous cell-permeable ceramide (C2-ceramide; 5 and 10 mM) significantly inhibited bone marrow CFU-E colony formation. This effect was reversed by the ceramide antagonist sphingosine-1-phosphate (S-1-P). Inhibition of CFU-E by rhgammaIFN, but not rhbetaIFN, was significantly reversed by S-1-P. rhEPO 10 U/mL reversed CFU-E inhibition by C2-ceramide 10 mM. Exposure of marrow cells to rhgammaIFN led to a 57% increase in ceramide content. The present study demonstrates that colony formation by human CFU-E is inhibited by endogenous and exogenous ceramide, and that inhibition by rhgammaIFN can be reversed by the ceramide antagonist S-1-P. Inhibition of CFU-E colony formation by ceramide and by are both reversed by high concentrations of rhEPO. These findings strongly suggest that ceramide mediates inhibition of human CFU-E colony formation by gammaIFN. PMID- 10390189 TI - Hematopoietic dynamics in grey collies. AB - Using Lomb periodogram analysis we have quantified variations in the peripheral neutrophil and platelet counts of the cyclical neutropenia animal model-the grey collie. We found that the amplitudes of the oscillations in these two cell lineages vary concomitantly. Further, the power spectrum and the shape of the oscillations in the absolute neutrophil counts vary together with the amplitude of the oscillations. As the amplitude of the oscillations increases, the height of the second subharmonic increases, giving rise to a distorted oscillation with two peaks per cycle. The particular dynamics of the absolute neutrophil counts can be reproduced by a combination of a delayed peripheral feedback, representing the peripheral control of granulopoiesis through granulocyte colony stimulating factor, together with a sinusoidal input representing an oscillatory input from the pluripotential stem cells to the granulocytic lineage. The same pluripotential stem cell input is probably responsible for the sinusoidal oscillations observed in the other cell lineages. PMID- 10390190 TI - Enzymatic and functional correction along with long-term enzyme secretion from transduced bone marrow hematopoietic stem/progenitor and stromal cells derived from patients with Fabry disease. AB - Fabry disease is a lysosomal storage disorder that is due to a deficiency in alpha-galactosidase A (alpha-gal A). Previously we have shown that a recombinant retrovirus synthesized for the transfer of the human alpha-gal A coding sequence was able to engineer enzymatic correction of the hydrolase deficiency in fibroblasts and lymphoblasts from Fabry patients. The corrected cells secreted alpha-gal A that was taken up and utilized by uncorrected bystander cells, thus demonstrating metabolic cooperativity. In separate experiments we used transduced murine bone marrow cells and successfully tested and quantitated this phenomenon in vivo. In the present studies, which were designed to bring this therapeutic approach closer to clinical utility, we establish that cells originating from the bone marrow of numerous Fabry patients and normal volunteers can be effectively transduced and that these target cells demonstrate metabolic cooperativity. Both isolated CD34+-enriched cells and long-term bone marrow culture cells, including nonadherent hematopoietic cells and adherent stromal cells, were transduced. The transferred gene generates increased intracellular alpha-gal A enzyme activity in these cells. Further, it causes functional correction of lipid accumulation and provides for long-term alpha-gal A secretion. Collectively, these results indicate that a multifaceted gene transfer approach to bone marrow cells may be of therapeutic benefit for patients with Fabry disease. PMID- 10390191 TI - Granulocyte colony-stimulating factor mobilized peripheral blood stem cells enter into G1 of the cell cycle and express higher levels of amphotropic retrovirus receptor mRNA. AB - We compared the cell cycle status and expression of mRNA for the amphotropic retroviral receptor in hematopoietic stem cells isolated from bone marrow and cytokine mobilized peripheral blood. CD34+ cells from six normal volunteers were enriched by immune selection from steady-state bone marrow and granulocyte colony stimulating factor (G-CSF) mobilized peripheral blood (10 microg/kg/day for 5 days). Cell cycle status of the phenotypically primitive CD34+CD38- hematopoietic stem cell population was analyzed using a four-color flow cytometry technique that distinguished the G0, G1, and S/IG2/M phases of the cell cycle. Semiquantitative reverse transcriptase-polymerase chain reaction was performed to measure mRNA expression of the amphotropic retroviral receptor. Peripheral blood hematopoietic stem cells had 2.6-fold more cells in the G1 phase of the cell cycle compared to steady-state bone marrow. Furthermore, lineage CD34+CD38- cells from G-CSF mobilized peripheral blood had a fourfold higher level of amphotropic retrovirus receptor mRNA. In conclusion, we found that CD34+ CD38- hematopoietic stem cells isolated from G-CSF mobilized peripheral blood differ from those isolated from steady-state bone marrow in that a significant proportion have entered the G1 phase of the cell cycle and express higher levels of amphotropic receptor mRNA. These biologic properties are consistent with the reported rapid recovery of hematopoietic function following transplantation with peripheral blood hematopoietic stem cells and make these cells a preferred target for retroviral-based gene transfer. PMID- 10390192 TI - NF-kappaB involvement in the activation of primary adult T-cell leukemia cells and its clinical implications. AB - The HTLV-I provirus-encoded Tax protein induces NF-kappaB in Tax-transfected Jurkat T cells or HTLVL-I- infected T cells in vitro. Tax induction of NF-kappaB is presumed to be involved in proliferation and activation of primary leukemia cells in vivo. Recent studies have demonstrated that NF-kappaB activities in human T cells are mediated by at least four c-Rel-related DNA binding proteins - p50, p55, p75 and p85. We examined the significance of NF-kappaB induction in primary adult T cell leukemia cells and the induction kinetics of each of the four NF-kappaB species. Marked NF-kappaB activity was detected using an electrophoretic mobility shift assay (EMSA) in the primary cells of patients with acute disease, but little activity was noted in the cells of chronic patients. NF kappaB activity was enhanced in a time-dependent manner in acute type cells cultured with mitogen-free medium; there was no induction of activity in chronic type cells. UV crosslinking demonstrated all four species of NFkappaB complex - high levels of p50 and lower levels of p55 and p75, in acute type cells; chronic type cells showed only the p50. As a control, normal resting T cells similarly showed only p50; control cells showed little change in activity when cultured without mitogenic stimulation, analogous to chronic type ATL. Northern blotting revealed enhancement of c-rel (encoding p85) and KBFI (encoding p50 and p55) expression in acute type cells during culture, while there was no significant enhancement of mRNAs in chronic type ATL cells or unstimulated normal T cells. Northern blotting also revealed that Tax is upregulated at the mRNA level in acute- but not chronic-type cells during culture. Expression of c-rel and KBF1 mRNAs in acute type cells appeared to be related to Tax mRNA expression. These results suggest that Tax is capable of inducing nuclear expression of all four NF kappaB species in primary ATL cells of acute type patients, with marked effects on p55, p75, and p85. Tax induction of NF-kappaB species is regulated, at least in part, at a pretranslational level involving increases in c-rel and KBF1 mRNA. PMID- 10390193 TI - Clonal heterogeneity of dendritic cells derived from patients with chronic myeloid leukemia and enhancement of their T-cells stimulatory activity by IFN alpha. AB - Adoptive immunotherapy in form of donor leukocyte infusions is effective in a significant number of patients with chronic myeloid leukemia (CML) that have relapsed after allogeneic bone marrow transplantation (BMT). However, the therapy is associated with clinically significant side effects such as graft-versus-host disease (GVHD) and bone marrow (BM) hypoplasia that may be avoided through the administration of T cells with specific antileukemic activity. Dendritic cells (DC) functioning as potent antigen presenting cells (APC) may play an important role in the generation of T cells with specificity against CML. We examined a subpopulation of CD1a+/CD14- DC generated in vitro from BM of normal subjects and patients with CML using granulocyte-macrophage colony-stimulating factor (GM CSF), tumor necrosis factor-alpha (TNF-alpha) and interleukin-4 (IL-4). These DC derived from both the BM of normal subjects and of patients with CML, differentiated and matured in culture in a similar way. However, DC derived from patients with CML, displayed decreased activity when tested with allogeneic T cells in a mixed lymphocyte reaction (MLR). Addition of interferon-alpha (IFN alpha) to DC cultures significantly upregulated the expression of major histocompatibility complex (MHC) molecules (class I and class II) and costimulatory molecules (B7.1 and B7.2) on DC from normal donors and CML patients. However, DC grown from CML patients required a higher concentration of IFN-alpha. IFN-alpha also significantly improved the capacity of CML DC to stimulate T-lymphocyte responses. Fluorescence in situ hybridization (FISH) showed that only some CD1a+/CD14- DC derived from BM of patients with CML expressed the bcr/abl fusion gene. Incubation with INF-alpha decreased the proportion of bcr/abl positive DC. PMID- 10390194 TI - Differentiation of antitumor-specific cytotoxic T lymphocytes from autologous tumor infiltrating lymphocytes in non-Hodgkin's lymphomas. AB - The present study describes a new culture protocol allowing the activation and proliferation of autologous tumor infiltrating T lymphocytes (TIL), and the generation of antitumor specific CTL in non-Hodgkin's lymphoma (NHL). Cells from eight patients with indolent NHL were used. We performed 3-week co-cultures of TIL with irradiated autologous malignant B cells in the presence of low doses of IL-1beta, IL-2 and IL-12. The proliferation, phenotype and cytotoxicity, and antitumor specificity of T cells recovered were studied. T-cell clonality was analyzed using TCRgamma gene rearrangement amplification by a multiplex PCR. Under these culture conditions, TIL proliferated, and the CD8+ T lymphocytes that were in a minority at the beginning of the culture increased dramatically in 6 out of 8 cases. In two cases, CD4+ T lymphocytes expanded. We showed that an oligoclonal selection of reactive T cells occurred in culture. Specific cytotoxicity developed against autologous malignant B cells in the 6 cases where there was an expansion of CD8+ T lymphocytes. Inhibition experiments performed with mAb directed against HLA class I and II molecules, CD4, CD8 and TCRgammadelta showed that the cytotoxic effector cells were CD8+ T lymphocytes probably expressing TCRalphabeta+. Cytokine secretion was analyzed in culture medium, and we detected significant levels of IFN-gamma, TNF-alpha, and IL-10 and no IL-4 (except in one case). Our results demonstrate that memory T cells from lymphoma patients can be amplified and differentiated into antitumor cytotoxic cells using a combination of the cytokines IL-1beta, IL-2, and IL-12 in association with non modified tumor cells. PMID- 10390195 TI - Apoptosis induced by human cytomegalovirus infection can be enhanced by cytokines to limit the spread of virus. AB - Fas-mediated apoptosis is one of the immune effector pathways leading to the elimination of virus infected cells. In vivo, apoptotic signals are delivered to virus infected cells by Fas-L and other cytokines secreted by specific T lymphocytes. Cellular immune response appears to be essential in prevention of human cytomegalovirus (HCMV) disease. We have hypothesized that HCMV infection might directly or indirectly result in upregulation of Fas receptor and in the presence of Fas ligand, lead to apoptosis of infected cells. We show that infection of human fibroblasts with HCMV is associated with upmodulation of Fas-R process that could be further potentiated by interferon (IFN-gamma). Using DNA agarose gel electrophoresis, terminal dideoxy transferase reaction, and annexin assay, we demonstrated that in a productive HCMV infection of human fibroblasts, loss of cell viability was not only due to virus-mediated cell lysis but also to due to apoptosis. IFN-gamma induced relative HCMV resistance and prevented loss in cell viability. In contrast, anti-Fas monoclonal antibody CH11, serving as Fas agonist, resulted in an accelerated loss in viability of infected cells. IFN gamma in combination with CH11 further increased the rate of apoptosis and compared to cultures with CH11 only, this effect was not restricted to only infected cells. While IFN-gamma did not affect the number of cells expressing immediate early antigen, it markedly reduced structural protein expression. IFN gamma in combination with CH11, decreased the expression of HCMV matrix protein pp65, reduced the amount of HCMV DNA and infectious virus produced. Our results are consistent with the theory that cells infected with HCMV can be eliminated by immune effector cells via Fas-mediated apoptosis. IFN-gamma, in addition to its intrinsic antiviral activity, primes HCMV infected cells to the action of Fas ligand and Fas-mediated apoptosis. PMID- 10390196 TI - The "G-CSF test": the response to a single dose of granulocyte colony-stimulating factor predicts mobilization of hemopoietic progenitors in patients with hematologic malignancies. AB - A significant proportion of patients with hematologic malignancies fail to mobilize sufficient hemopoietic progenitor cells (HPC), thereby restricting wider application of autologous transplantation. It would be of considerable use to develop a test that could be used prospectively to assess an individual patient's capacity to mobilize HPC. Twenty-two patients with lymphoma, myeloma, and chronic lymphocytic leukemia were given a single dose of 12 microg/kg SC of granulocyte colony-stimulating factor (G-CSF). Blood colony-forming unit granulocyte macrophage (CFU-GM) and CD34+ cells were scored prior to the test dose, and 72, 96, and 120 hours later. The patients were then mobilized with a standard cyclophosphamide and G-CSF regimen and had blood stem cells harvested. Patients were categorized as good, poor, or intermediate mobilizers on the basis of the CFU-GM/CD34+ cell harvest content and the number of aphereses required to reach established threshold counts. The outcome of cyclophosphamide/G-CSF mobilization was correlated with the response to the test dose of G-CSF. Good mobilizers had significantly higher peak CFU-GM values and CFU-GM increment in response to the test dose of G-CSF compared to intermediate and poor mobilizers. A peak CFU-GM count of > or = 250/mL identified the good mobilizers; conversely, all poor mobilizers had a peak CFU-GM count of <102/mL. An increment in CD34+ cells counts of > or = 2.5/microL was only observed in good mobilizers. The "G-CSF" test is a reliable test that can be used successfully for the assessment of mobilizable HPC in patients with hematologic malignancies. It can also be used to stratify patients enrolled in trials of mobilizing agents. PMID- 10390197 TI - Engraftment of marrow allografts treated with Campath-1 monoclonal antibodies. AB - We have analyzed the factors associated with engraftment in 216 recipients of T cell depleted allogeneic HLA identical sibling marrow transplants using Campath 1 monoclonal antihuman lymphocyte (CD52) antibodies. The patient population consisted of 168 patients with hematologic malignancies, 26 with severe aplastic anemia (SAA), and 22 with hemoglobinopathies, half of whom received marrow treated in vitro with Campath-1M (IgM) and half received marrow with Campath-1G (IgG2b isotype). Patients with durable engraftment had fast hematopoietic recovery: SAA patients reached ANC > 0.5 x 10(6)/L on Day 14; those with leukemia attained ANC > 0.5 x 10(6)/L on Days 18, 17, and 15 for ANLL, ALL and CML respectively, while patients with thalasemia reached ANC > 0.5 x 10(6)/L on Day 21. Overall, 24 patients (17 with leukemia, 4 with SAA, and 3 with thalassemia) suffered graft failure: 10 patients (all grafted with Campath-1M) rejected their grafts, while 14 others (9 grafted with Campath-1M, and 5 with 1G isotype) never engrafted (p = 0.009). Multivariate analysis revealed that neither pretransplant protocol, nor stage of disease or type of antibody used, donor sex and ABO match had any impact on engraftment. The variables favorably associated with engraftment were older age (p = 0.030, RR = 1.016) and CFU-GM number (p = 0.013, RR = 1.001). Patients with ANLL or SAA had a better chance to engraft (p = 0.027, RR = 1.400; and p = 0.003, RR = 2.677, respectively) compared to patients with thalassemia (p = 0.001, RR = 0.551). A higher concentration of Campath-1 antibody in vitro and in vivo adversely affected engraftment. Our data show that satisfactory engraftment can be achieved in patients transplanted with Campath-1 treated marrow allografts. However, despite the measures undertaken to prevent rejection, graft failure still poses a problem. Further pretransplant immunosuppression and perhaps more selective T-cell depletion may reduce the increased graft failure in these patients. PMID- 10390198 TI - Effectiveness of rotor off fraction in allogeneic murine bone marrow transplantation with complete disparity of major histocompatibility. AB - Counterflow centrifugal elutriation (CCE) has been a highly efficient physical method for separating T cells from bone marrow (BM) without impairing cell function and yield. To investigate the usefulness of CCE, the hematopoietic potential as well as the level of T cell contamination in rotor-off (R/O) fraction of BM was studied using a murine bone marrow transplantation (BMT) model [C3H/He (H-2k)-->BALB/C (H-2d)]. The total recovery of cells after CCE procedure was 71.4%. Morphologically, R/O fraction contained abundant mononuclear cells and a few lymphocytes. The numbers of colony forming unit for granulocyte/monocyte (CFU-GM), Sca-1+ cells, and T cells were compared among four fractions of CCE (fractions at flow rate of 17, 25, 28 mL/min, and R/O fraction). The number of CFU-GM per 10(5) nucleated cells in each fraction were significantly higher in R/O fraction (331.3 +/- 34.4) compared to unfractionated marrow (UM) (21.1 +/- 1.3) and fraction of 17 mL/min (FR 17) (23.7 +/- 2.2 ) (chi2 = 0.0044). Neither fraction of 25 mL/min (FR 25) nor fraction of 28 mL/min (FR 28) contained CFU-GM colonies. The concentration of Sca-1+ cells in R/O fraction was significantly higher (1.96-fold) than UM (p < 0.05), and 80.0 +/- 10.1% of Sca-1+ cells in UM were recovered in R/O fraction; 88.1% of Thy-1.2+ T cells were eliminated in R/O fraction (p < 0.05). Mice receiving UM after lethal irradiation (875cGy) suffered from severe graft-versus-host disease (GVHD) and all five died within 7 days after BMT procedure (Group A). Of interest, mice receiving mixture of R/O fraction with lymphocyte-rich fraction (FR 25 plus FR 28) to equalize T cell number as UM, developed severe GVHD and four out of five died (probability of survival; 20%) (Group B). Mice receiving R/O fraction had mild GVHD and four out of five survived for at least 90 days (probability of survival; 80%) (Group C). In group C, probability of survival (p = 0.0006) was higher, and severity of GVHD (p = 0.0043) and progression rate of GVHD (p = 0.02) was lower. In conclusion, the elutriated R/O fraction cells of BM have the advantages of stable engraftment and tolerable GVHD in murine allogeneic BMT with complete major histocompatibility disparity. This could be directly applicable to patients with high risk of GVHD and graft failure in upcoming clinical trials. PMID- 10390199 TI - Why metronidazole is active against both bacteria and parasites. PMID- 10390200 TI - Salt-resistant alpha-helical cationic antimicrobial peptides. AB - Analogues based on the insect cecropin-bee melittin hybrid peptide (CEME) were studied and analyzed for activity and salt resistance. The new variants were designed to have an increase in amphipathic alpha-helical content (CP29 and CP26) and in overall positive charge (CP26). The alpha-helicity of these peptides was demonstrated by circular dichroism spectroscopy in the presence of liposomes. CP29 was shown to have activity against gram-negative bacteria that was similar to or better than those of the parent peptides, and CP26 had similar activity. CP29 had cytoplasmic membrane permeabilization activity, as assessed by the unmasking of cytoplasmic beta-galactosidase, similar to that of CEME and its more positively charged derivative named CEMA, whereas CP26 was substantially less effective. The activity of the peptides was not greatly attenuated by an uncoupler of membrane potential, carbonyl cyanide-m-chlorophenylhydrazone. The tryptophan residue in position 2 was shown to be necessary for interaction with cell membranes, as demonstrated by a complete lack of activity in the peptide CP208. Peptides CP29, CEME, and CEMA were resistant to antagonism by 0.1 to 0.3 M NaCl; however, CP26 was resistant to antagonism only by up to 160 mM NaCl. The peptides were generally more antagonized by 3 and 5 mM Mg2+ and by the polyanion alginate. It appeared that the positively charged C terminus in CP26 altered its ability to permeabilize the cytoplasmic membrane of Escherichia coli, although CP26 maintained its ability to kill gram-negative bacteria. These peptides are potential candidates for future therapeutic drugs. PMID- 10390201 TI - Prospective evaluation of the effect of an aminoglycoside dosing regimen on rates of observed nephrotoxicity and ototoxicity. AB - The nephrotoxicity and ototoxicity associated with once-daily versus twice-daily administration of aminoglycosides was assessed in patients with suspected or proven gram-negative bacterial infections in a randomized, double-blind clinical trial. Patients who received therapy for >/=72 h were evaluated for toxicity. Patients also received concomitant antibiotics as deemed necessary for treatment of their infection. Plasma aminoglycoside concentrations, prospective aminoglycoside dosage adjustment, and serial audiologic and renal status evaluations were performed. The probability of occurrence of a nephrotoxic event and its relationship to doses and daily aminoglycoside exposure served as the main outcome measurement. One hundred twenty-three patients were enrolled in the study, with 83 patients receiving therapy for at least 72 h. For 74 patients plasma aminoglycoside concentrations were available for analysis, and the patients formed the group evaluable for toxicity. The primary infectious diagnosis for the patients who were enrolled in the study were bacteremia or sepsis, respiratory infections, skin and soft tissue infections, or urosepsis or pyelonephritis. Of the 74 patients evaluable for toxicity, 39 received doses twice daily and 35 received doses once daily and a placebo 12 h later. Nephrotoxicity occurred in 6 of 39 (15.4%) patients who received aminoglycosides twice daily and 0 of 35 patients who received aminoglycosides once daily. The schedule of aminoglycoside administration, concomitant use of vancomycin, and daily area under the plasma concentration-time curve (AUC) for the aminoglycosides were found to be significant predictors of nephrotoxicity by multivariate logistic regression analysis (P /=1.0 mg/liter, little killing occurred over 48 h. A sigmoid dose-response model indicated that the area under the curve/MIC ratio was strongly related to the log change in viable count at 24 and 48 h and to the AUBKC. These data indicate that moxifloxacin may have a role in management of S. pneumoniae infection. PMID- 10390204 TI - Identification and analysis of the balhimycin biosynthetic gene cluster and its use for manipulating glycopeptide biosynthesis in Amycolatopsis mediterranei DSM5908. AB - Seven complete genes and one incomplete gene for the biosynthesis of the glycopeptide antibiotic balhimycin were isolated from the producer, Amycolatopsis mediterranei DSM5908, by a reverse-cloning approach and characterized. Using oligonucleotides derived from glycosyltransferase sequences, a 900-bp glycosyltransferase gene fragment was amplified and used to identify a DNA fragment of 9,882 bp. Of the identified open reading frames, three (oxyA to -C) showed significant sequence similarities to cytochrome P450 monooxygenases and one (bhaA) showed similarities to halogenase, and the genes bgtfA to -C showed similarities to glycosyltransferases. Glycopeptide biosynthetic mutants were created by gene inactivation experiments eliminating oxygenase and glycosyltransferase functions. Inactivation of the oxygenase gene(s) resulted in a balhimycin mutant (SP1-1) which was not able to synthesize an antibiotically active compound. Structural analysis by high-performance liquid chromatography mass spectrometry, fragmentation studies, and amino acid analysis demonstrated that these oxygenases are involved in the coupling of the aromatic side chains of the unusual heptapeptide. Mutant strain HD1, created by inactivation of the glycosyltransferase gene bgtfB, produced at least four different compounds which were not glycosylated but still antibiotically active. PMID- 10390205 TI - DNA cleavage activities of Staphylococcus aureus gyrase and topoisomerase IV stimulated by quinolones and 2-pyridones. AB - We have cloned Staphylococcus aureus DNA gyrase and topoisomerase IV and expressed them in Escherichia coli as polyhistidine-tagged proteins to facilitate purification and eliminate contamination by host enzymes. The enzyme preparations had specific activities similar to previously reported values. Potassium glutamate (K-Glu) stimulated the drug-induced DNA cleavage activity and was optimal between 100 and 200 mM for gyrase and peaked at 100 mM for topoisomerase IV. Higher concentrations of K-Glu inhibited the cleavage activities of both enzymes. Using a common buffer system containing 100 mM K-Glu, we tested the enzyme-mediated DNA cleavage activities of both gyrase and topoisomerase IV with oxolinic acid, norfloxacin, ciprofloxacin, trovafloxacin, clinafloxacin, and the 2-pyridone ABT-719. As expected, all drugs tested demonstrated greater potency against topoisomerase IV than against gyrase. In addition, cleavage activity was found to correlate well with antibacterial activity. PMID- 10390206 TI - Cloning and characterization of a gene, pbpF, encoding a new penicillin-binding protein, PBP2B, in Staphylococcus aureus. AB - A previously unrecognized penicillin binding protein (PBP) gene, pbpF, was identified in Staphylococcus aureus. This gene encodes a protein of 691 amino acid residues with an estimated molecular mass of 78 kDa. The molecular mass is very close to that of S. aureus PBP2 (81 kDa), and the protein is tentatively named PBP2B. PBP2B has three motifs, SSVK, SSN, and KTG, that can be found in PBPs and beta-lactamases. Recombinant PBP2B (rPBP2B), which lacks a putative signal peptide at the N terminus and has a histidine tag at the C terminus, was expressed in Escherichia coli. The purified rPBP2B was shown to have penicillin binding activity. A protein band was detected from S. aureus membrane fraction by immunoblotting with anti-rPBP2B serum. Also, penicillin binding activity of the protein immunoprecipitated with anti-rPBP2B serum was detected. These results suggest the presence of PBP2B in S. aureus cell membrane that covalently binds penicillin. The internal region of pbpF and PBP2B protein were found in all 12 S. aureus strains tested by PCR and immunoblotting. PMID- 10390208 TI - Beta-lactamase production by oral anaerobic gram-negative species in infants in relation to previous antimicrobial therapy. AB - The frequency of beta-lactamase production in gram-negative bacteria has increased considerably during recent years. In this study, beta-lactamase production by oral anaerobic gram-negative rods isolated from saliva was longitudinally examined for 44 Caucasian infants at the ages of 2, 6, and 12 months in relation to their documented exposure to antibiotics. Isolates showing decreased susceptibility to penicillin G (1 microg/ml) were examined for beta lactamase production by using a chromogenic cephalosporin disk test. beta Lactamase-positive, gram-negative anaerobic species were found in 11, 55, and 89% of each age group, respectively. beta-Lactamase production was most frequent among organisms of the Prevotella melaninogenica group. At 12 months, 73% of the infants harbored beta-lactamase-producing members of the P. melaninogenica group, 55% had nonpigmented Prevotella species, 25% had Porphyromonas catoniae, 23% had Fusobacterium nucleatum, and 5% had Capnocytophaga species. Several beta lactamase-producing species could be simultaneously found in the infants' mouths. The presence of beta-lactamase-producing species was significantly associated with the infants' exposure to antibiotics through antimicrobial treatments given to the infants and/or their mothers. PMID- 10390209 TI - Antimicrobial susceptibility and composition of microcosm dental plaques supplemented with sucrose. AB - The aims of this study were to evaluate the effects of repeated chlorhexidine gluconate (CHG) pulsing on the viability and bacterial composition of microcosm dental plaques derived from human saliva. The biofilms were grown on bovine enamel discs in a constant-depth film fermentor fed with an artificial saliva which was supplemented thrice daily with sucrose. The microcosm plaques had total viable anaerobic counts of 5 x 10(8) CFU per mm2 and consisted of 12% Actinomyces spp., 85% streptococci, and 0.2% Veillonella spp. When pulsed twice daily with 0.2% CHG, there was an immediate 1.3-log10 reduction in the total viable (anaerobic) count. However, as pulsing continued, the viable counts recovered, and after 4 days, the anaerobic count reached its pre-CHG-pulsing level, although the bacterial composition of the biofilms had changed. The results of this study show that twice-daily pulsing with 0.2% CHG over a 4-day period was ineffective at reducing the total anaerobic viable count of the biofilms but did alter their bacterial composition. PMID- 10390207 TI - Cloning and characterization of blaVIM, a new integron-borne metallo-beta lactamase gene from a Pseudomonas aeruginosa clinical isolate. AB - Production of a metallo-beta-lactamase activity was detected in a carbapenem resistant Pseudomonas aeruginosa clinical isolate (isolate VR-143/97) from an Italian inpatient at the Verona University Hospital (northern Italy). The metallo beta-lactamase determinant was isolated from a genomic library of VR-143/97, constructed in an Escherichia coli plasmid vector, by screening for clones with reduced susceptibility to imipenem. Sequencing of the cloned gene revealed that it encoded a new class B beta-lactamase that was named VIM-1. At the sequence level VIM-1 was rather divergent from the other class B enzymes (16.4 to 38.7% identity), overall being more similar to members of subclass B1 including the beta-lactamase II of Bacillus cereus (Bc-II), the Bacteroides fragilis CcrA, the Chryseobacterium meningosepticum BlaB, and the cassette-encoded IMP-1 enzymes. Among these, VIM-1 showed the highest degree of similarity to Bc-II. Similarly to blaIMP, blaVIM was also found to be carried on a gene cassette inserted into a class 1 integron. The blaVIM-containing integron was located on the chromosome of P. aeruginosa VR-143/97, and the metallo-beta-lactamase-encoding determinant was not transferable to E. coli by conjugation. Expression of the integron-borne blaVIM gene in E. coli resulted in a significant decrease in susceptibility to a broad array of beta-lactams (ampicillin, carbenicillin, piperacillin, mezlocillin, cefotaxime, cefoxitin, ceftazidime, cefoperazone, cefepime, and carbapenems), revealing a very broad substrate specificity of the VIM-1 enzyme. PMID- 10390210 TI - Use of real-time PCR and fluorimetry to detect lamivudine resistance-associated mutations in hepatitis B virus. AB - Very rapid amplification of DNA by PCR in small volumes can be continuously monitored by the detection of the binding of probes with a rapid cycler with built-in fluorometric detection. Primers were designed to amplify approximately 100 bp of the polymerase gene of hepatitis B virus (HBV) spanning codon 550, where mutations associated with resistance to lamivudine invariably occur. Four hybridization probes were synthesized: one was 3' labelled with fluorescein and hybridized upstream of codon 550. The others were 5' labelled with Cy5 and 3' labelled with biotin and spanned codon 550. The Cy5-labelled oligonucleotides contained either wild-type (ATG) or mutant (GTG or ATT) sequences. A Cy5-labelled probe and either the fluorescein-labelled probe or Sybr Green 1 (a compound that fluoresces when bound to double-stranded DNA) were included in each PCR. After completion of the amplification by using a LightCycler (Idaho Technology), the temperature at which the Cy5 probe melted from the product was determined in a melt program that took ca. 3 min. Pre- and posttreatment samples from eight patients (five chronic and three transplant) who failed lamivudine treatment were amplified, and the presence of mutations in codon 550 was determined by ABI sequencing and by using the LightCycler; in some cases PCR products were also cloned, and multiple clones were sequenced. Concordant results were obtained in all cases. We found the LightCycler to be better at resolving the sequences of genomic mixtures; for example, two samples showed a sequence at codon 550 of (A/G)T(G/T), which was found by fluorimetry to be mixtures of GTG and ATT but no ATG, and this finding was confirmed by the sequencing of clones. However, this approach was not more sensitive than population sequencing for the detection of the presence of mixtures. Overall, this pilot study has demonstrated an approach that could be an extremely rapid and economical method for the detection of lamivudine resistance-associated mutations in HBV. PMID- 10390211 TI - Efficacy of locally delivered polyclonal immunoglobulin against Pseudomonas aeruginosa peritonitis in a murine model. AB - Infectious peritonitis results from bacterial contamination of the abdominal cavity. Conventional antibiotic treatment is complicated both by the emergence of antibiotic-resistant bacteria and by increased patient populations intrinsically at risk for nosocomial infections. To complement antibiotic therapies, the efficacy of direct, locally applied pooled human immunoglobulin G (IgG) was assessed in a murine model (strains CF-1, CD-1, and CFW) of peritonitis caused by intraperitoneal inoculations of 10(6) or 10(7) CFU of Pseudomonas aeruginosa (strains IFO-3455, M-2, and MSRI-7072). Various doses of IgG (0.005 to 10 mg/mouse) administered intraperitoneally simultaneously with local bacterial challenge significantly increased survival in a dose-dependent manner. Local intraperitoneal application of 10 mg of IgG increased animal survival independent of either the P. aeruginosa or the murine strains used. A local dose of 10 mg of IgG administered up to 6 h prophylactically or at the time of bacterial challenge resulted in 100% survival. Therapeutic 10-mg IgG treatment given up to 12 h postinfection also significantly increased survival. Human IgG administered to the mouse peritoneal cavity was rapidly detected systemically in serum. Additionally, administered IgG in peritoneal lavage fluid samples actively opsonized and decreased the bacterial burden via phagocytosis at 2 and 4 h post bacterial challenge. Tissue microbial quantification studies showed that 1.0 mg of locally applied IgG significantly reduced the bacterial burden in the liver, peritoneal cavity, and blood and correlated with reduced levels of interleukin-6 in serum. PMID- 10390212 TI - Safety and efficacy of intravenous zanamivir in preventing experimental human influenza A virus infection. AB - Zanamivir is a potent inhibitor of influenza A and B virus neuraminidases and is active topically in experimental and natural human influenza. We conducted this double-blinded, placebo-controlled study to evaluate the safety and efficacy of intravenously administered zanamivir. Susceptible volunteers were randomized to receive either saline or zanamivir (600 mg) intravenously twice daily for 5 days beginning 4 h prior to intranasal inoculation with approximately 10(5) 50% tissue culture infectious doses (TCID50) of influenza A/Texas/36/91 (H1N1) virus. Reductions in the frequency of viral shedding (0% versus 100% in placebo, P < 0.005) and seroconversion (14% versus 100% in placebo, P < 0.005) and decreases in viral titer areas under the curve (0 versus 11.6 [median] log10 TCID50. day/ml in placebo, P < 0.005) were observed in the zanamivir group, as were reductions in fever (14% versus 88% in placebo, P < 0.05), upper respiratory tract illness (0% versus 100% in placebo, P < 0.005), total symptom scores (1 versus 44 [median] in placebo, P < 0.005), and nasal-discharge weight (3.9 g versus 17.5 g [median] in placebo, P < 0.005). Zanamivir was detectable in nasal lavage samples collected on days 2 and 4 (unadjusted median concentrations, 10.5 and 12.0 ng/ml of nasal wash, respectively). This study demonstrates that intravenously administered zanamivir is distributed to the respiratory mucosa and is protective against infection and illness following experimental human influenza A virus inoculation. PMID- 10390214 TI - Topoisomerase sequences of coagulase-negative staphylococcal isolates resistant to ciprofloxacin or trovafloxacin. AB - Coagulase-negative staphylococcal isolates (n = 188) were screened for susceptibility to oxacillin, ciprofloxacin, and trovafloxacin, a new fluoroquinolone. At an oxacillin concentration of >/=4 microg/ml, 43% were methicillin resistant; of these, 70% were ciprofloxacin resistant (MIC, >/=4 microg/ml). Of the methicillin-resistant, ciprofloxacin-resistant isolates, 46% were susceptible to /=8 microg/ml) and increased trovafloxacin MICs (0.25 to 2 microg/ml) could be conferred by the combined presence of single mutations in each gyrA and grlA gene. Trovafloxacin MICs of >/=8 microg/ml also occurred, but these required an additional mutation in grlA. PMID- 10390213 TI - Multiple resistant phenotypes of Candida albicans coexist during episodes of oropharyngeal candidiasis in human immunodeficiency virus-infected patients. AB - Mechanisms of resistance to azoles in Candida albicans, the main etiologic agent of oropharyngeal candidiasis (OPC), include alterations in the target enzyme (lanosterol demethylase) and increased efflux of drug. Previous studies on mechanisms of resistance have been limited by the fact that only a single isolate from each OPC episode was available for study. Multiple isolates from each OPC episode were evaluated with oral samples plated in CHROMagar Candida with and without fluconazole to maximize detection of resistant yeasts. A total of 101 isolates from each of three serial episodes of OPC from four different patients were evaluated. Decreasing geometric means of fluconazole MICs with serial episodes of infection were detected in the four patients. However, 8-fold or larger (up to 32-fold) differences in fluconazole MICs were detected within isolates recovered at the same time point in 7 of 12 episodes. Strain identity was analyzed by DNA typing techniques and indicated that isolates from each patient represented mainly isogenic strains, but differed among patients. A Northern blot technique was used to monitor expression of ERG11 (encoding lanosterol demethylase) and genes coding for efflux pumps. This analysis revealed that clinical isolates obtained from the same patient and episode were phenotypically heterogeneous in their patterns of expression of these genes involved in fluconazole resistance. These results demonstrate the complexity of the distribution of the molecular mechanisms of antifungal drug resistance and indicate that different subpopulations of yeasts may coexist at a given time in the same patient and may develop resistance through different mechanisms. PMID- 10390215 TI - Effective treatment of acute and chronic murine tuberculosis with liposome encapsulated clofazimine. AB - The therapeutic efficacy of liposomal clofazimine (L-CLF) was studied in mice infected with Mycobacterium tuberculosis Erdman. Groups of mice were treated with either free clofazimine (F-CLF), L-CLF, or empty liposomes twice a week for five treatments beginning on day 1 (acute), day 21 (established), or day 90 (chronic) postinfection. One day after the last treatment, the numbers of CFU of M. tuberculosis in the spleen, liver, and lungs were determined. F-CLF at the maximum tolerated dose of 5 mg/kg of body weight was ineffective; however, 10 fold-higher doses of L-CLF demonstrated a dose response with significant CFU reduction in all tissues without any toxic effects. In acutely infected mice, 50 mg of L-CLF/kg reduced CFU 2 to 3 log units in all three organs. In established or chronic infection, treated mice showed no detectable CFU in the spleen or liver and 1- to 2-log-unit reduction in the lungs. A second series of L-CLF treatments cleared M. tuberculosis in all three tissues. L-CLF appears to be bactericidal in the liver and spleen, which remained negative for M. tuberculosis growth for 2 months. Thus, L-CLF could be useful in the treatment of tuberculosis. PMID- 10390216 TI - Biochemical-genetic analysis and distribution of FAR-1, a class A beta-lactamase from Nocardia farcinica. AB - From genomic DNA of the clinical isolate Nocardia farcinica VIC, a 1. 6-kb Sau3AI fragment was cloned and expressed in Escherichia coli JM109. The recombinant strain expressed a beta-lactamase (pI, 4.6), FAR-1, which conferred high levels of resistance to amoxicillin, piperacillin, ticarcillin, and cephalothin. The hydrolysis constants (kcat, Km, Ki, and 50% inhibitory concentration) confirmed the MIC results and showed that FAR-1 activity is inhibited by clavulanic acid and at a low level by tazobactam and sulbactam. Moreover, FAR-1 beta-lactamase hydrolyzes aztreonam (at a low level) without significant activity against ceftazidime, cefotaxime and imipenem. FAR-1 mature protein of molecular mass ca 32 kDa, has less than 60% amino acid identity with any other class A beta lactamases, being most closely related to PEN-A from Burkholderia cepacia (52%). A blaFAR-1-like gene was found in all studied N. farcinica strains, underlining the constitutive origin of this gene. PMID- 10390217 TI - Comparative efficacies of antibiotics in a rat model of meningoencephalitis due to Listeria monocytogenes. AB - The antibacterial activities of amoxicillin-gentamicin, trovafloxacin, trimethoprim-sulfamethoxazole (TMP-SMX) and the combination of trovafloxacin with TMP-SMX were compared in a model of meningoencephalitis due to Listeria monocytogenes in infant rats. At 22 h after intracisternal infection, the cerebrospinal fluid was cultured to document meningitis, and the treatment was started. Treatment was instituted for 48 h, and efficacy was evaluated 24 h after administration of the last dose. All tested treatment regimens exhibited significant activities in brain, liver, and blood compared to infected rats receiving saline (P < 0.001). In the brain, amoxicillin plus gentamicin was more active than all of the other regimens, and trovafloxacin was more active than TMP SMX (bacterial titers of 4.1 +/- 0.5 log10 CFU/ml for amoxicillin-gentamicin, 5.0 +/- 0.4 log10 CFU/ml for trovafloxacin, and 5.8 +/- 0.5 log10 CFU/ml for TMP-SMX; P < 0.05). In liver, amoxicillin-gentamicin and trovafloxacin were similarly active (2.8 +/- 0.8 and 2.7 +/- 0.8 log10 CFU/ml, respectively) but more active than TMP-SMX (4.4 +/- 0. 6 log10 CFU/ml; P < 0.05). The combination of trovafloxacin with TMP-SMX did not alter the antibacterial effect in the brain, but it did reduce the effect of trovafloxacin in the liver. Amoxicillin gentamicin was the most active therapy in this study, but the activity of trovafloxacin suggests that further studies with this drug for the treatment of Listeria infections may be warranted. PMID- 10390218 TI - Molecular characterization of TEM-59 (IRT-17), a novel inhibitor-resistant TEM derived beta-lactamase in a clinical isolate of Klebsiella oxytoca. AB - A clinical isolate of Klebsiella oxytoca (Kox 443) was found to have a low-level resistance to broad-spectrum penicillins (MICs of amoxicillin and ticarcillin, 256 and 32 microg/ml, respectively), without substantial potentiation by 2 microg of clavulanic acid per ml (amoxicillin- and ticarcillin-clavulanate, 128 and 8 microg/ml, respectively), while being fully susceptible to cephalosporins and other beta-lactam antibiotics. These resistances were carried by a ca. 50-kb conjugative plasmid that encodes a single beta-lactamase with a pI of 5.6. Compared to TEM-2, this enzyme exhibited a 3- to 30-fold higher Km and a decreased maximal hydrolysis rate for beta-lactams; higher concentrations of suicide inactivators (5- to 500-fold higher concentrations giving a 50% reduction in hydrolysis) were required for inhibition. Nucleotide sequence analysis revealed identity between the blaTEM gene of Kox 443 and the blaTEM-2 gene, except for a single A-to-G change at position 590, leading to the amino acid change from Ser-130 Gly. This mutation has not been reported previously in the TEM type beta-lactamases produced by clinical strains, and the novel enzyme was called TEM-59 (alternative name IRT-17). This is the first description of an inhibitor-resistant TEM-derived enzyme in the species K. oxytoca. PMID- 10390219 TI - Nonactin biosynthesis: the product of nonS catalyzes the formation of the furan ring of nonactic acid. AB - Nonactin is the parent compound of a group of ionophore antibiotics, known as the macrotetrolides, produced by Streptomyces griseus subsp. griseus ETH A7796. Nonactin is a significant compound because of its inhibitory effects on the P170 glycoprotein-mediated efflux of chemotherapeutic agents in multiple-drug resistant cancer cells. Nonactin is also significant in that it is a highly atypical polyketide. Very little is presently known about the genes of the nonactin biosynthesis cluster. In this paper we describe our efforts to establish a connection between the product of a gene from the nonactin biosynthesis cluster and a known biochemical transformation in nonactin biosynthesis. Nonactate synthase is the enzyme which catalyzes the formation of nonactic acid from an acyclic precursor in nonactin biosynthesis. We have synthesized the substrate for this enzyme and have detected the in vitro cyclization activity of the substrate in cell-free preparations of S. griseus subsp. griseus ETH A7796. Previous studies by R. Plater and J. A. Robinson (Gene 112:117-122, 1992) had suggested, based on sequence homology, that the product of a partial open reading frame found close to the tetranactin resistance gene of S. griseus could be the nonactate synthase. We have therefore cloned, sequenced, and heterologously expressed this full gene (nonS), and we have shown that the gene product, NonS, does indeed catalyze the formation of the furan ring of nonactic acid as hypothesized. PMID- 10390220 TI - Roles of beta-lactamases and porins in activities of carbapenems and cephalosporins against Klebsiella pneumoniae. AB - Two clinical isolates of extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae were noted to be less susceptible than expected to imipenem. Both were missing outer membrane proteins that serve as channels for antibiotic entry. The role of beta-lactamase in resistance was investigated by eliminating the original ESBL and introducing plasmids encoding various ESBLs and AmpC beta-lactamase types, by studying the effect of an increased inoculum, and by evaluating interactions with beta-lactamase inhibitors. The contribution of porin deficiency was investigated by restoring a functional ompK36 gene on a plasmid. Plasmids encoding AmpC-type beta-lactamases provided resistance to imipenem (up to 64 microg/ml) and meropenem (up to 16 microg/ml) in strains deficient in porins. Carbapenem resistance showed little inoculum effect, was not affected by clavulanate but was blocked by BRL 42715, and was diminished if OmpK36 porin was restored. Plasmids encoding TEM- and SHV-type ESBLs conferred resistance to cefepime and cefpirome, as well as to earlier oxyimino-beta lactams. This resistance was magnified with an increased inoculum, was blocked by clavulanate, and was also lowered by OmpK36 porin restoration. In addition, SHV-2 beta-lactamase had a small effect on carbapenem resistance (imipenem MIC, 4 microg/ml, increasing to 16 microg/ml with a higher inoculum) when porins were absent. In K. pneumoniae porin loss can thus augment resistance provided either by TEM- or SHV-type ESBLs or by plasmid-mediated AmpC enzymes to include the latest oxyimino-beta-lactams and carbapenems. PMID- 10390221 TI - Sequence diversity of the reverse transcriptase of human immunodeficiency virus type 1 from untreated Brazilian individuals. AB - The presence of human immunodeficiency virus type 1 (HIV-1) bearing mutations resistant to nucleosidic inhibitors of the viral reverse transcriptase (RT) derived from HIV-seropositive asymptomatic and untreated volunteer blood donors was examined. The RT amplicons of 32 specimens were analyzed by using a reverse hybridization line probe assay technique that detects resistance against zidovudine (3'-azido-3'-deoxythymidine [AZT], didanosine (2',3'-dideoxyinosine [ddI], zalcitabine (2',3'-dideoxycytidine [ddC]), and lamivudine ((-)-beta-L 2',3'-dideoxy-3'-thiacytidine [3TC]) at amino acid positions 41, 69, 70, 74, 184, and 215 of the HIV RT. One sample (brp004, subtype B) showed an AZT resistance secondary mutation at position K70R. Fifteen specimens revealed one or more sites of nonreactivity to both wild-type- and mutant-specific probes (dual nonreactivity). Samples were also submitted to RT direct sequencing and phylogenetic analysis. Nine of 32 specimens belonged to non-B subtypes (C, D, F, and F/B or B/F mosaics). Three of these non-B isolates, named brp004, brp063, and brp069, revealed three other relevant AZT resistance mutations-a T215F mutation and two M41L mutations, respectively-hidden by the nonreactivity to line probe assay strips on the respective codon regions. The isolate brp004 also carried a D67N AZT resistance mutation revealed by direct sequencing. No nonnucleosidic RT inhibitor-resistant mutation was found. The analysis revealed a frequency of 2.26 x 10(-4) mutations per nucleotide for independent samples related to RT resistance. These findings emphasize the magnitude of naturally occurring reservoirs of drug-resistant virus among untreated HIV-1-positive individuals in Brazil. PMID- 10390222 TI - Structural alterations in the translational attenuator of constitutively expressed ermC genes. AB - Sequence deletions of 16, 59, and 111 bp as well as a tandem duplication of 272 bp with respect to the corresponding sequence of pT48 were identified in the regulatory regions of constitutively expressed ermC genes. Constitutive ermC gene expression as a consequence of these structural alterations is based on either the prevention of the formation of mRNA secondary structures in the translational attenuator or the preferential formation of those mRNA secondary structures which do not interfere with the translation of the ermC transcripts. A model for the development of sequence deletions in the ermC translational attenuator by homologous recombination is presented and experimentally tested by in vitro selection of constitutively expressed mutants in staphylococcal strains deficient and proficient in homologous recombination. PMID- 10390223 TI - Safety and pharmacokinetics of amprenavir (141W94), a human immunodeficiency virus (HIV) type 1 protease inhibitor, following oral administration of single doses to HIV-infected adults. AB - We conducted a double-blind, placebo-controlled, parallel, dose-escalation trial to evaluate the pharmacokinetics and safety of single, oral doses of amprenavir (141W94; formerly VX-478), a potent inhibitor of human immunodeficiency virus (HIV) type 1 protease, administered as hard gelatin capsules in 12 HIV-infected subjects. The doses of amprenavir evaluated were 150, 300, 600, 900, and 1,200 mg. Amprenavir was rapidly absorbed, with the time to maximum concentration occurring within 1 to 2 h after dosing. On the basis of power model analysis, the increase in the maximum concentration of amprenavir in plasma (Cmax) was less than dose proportional, and the increase in the area under the concentration-time curve from time zero to infinity (AUC0-infinity) was greater than dose proportional; mean slopes (with 90% confidence intervals) were 1.25 (1.16 to 1.35) and 0.78 (0.78 to 0.86) for AUC0-infinity and Cmax, respectively. Amprenavir was eliminated slowly, with a terminal-phase half-life of 8 h. A second study was conducted to determine the bioavailability of the hard gelatin capsule relative to that of a subsequently developed soft gelatin capsule. The capsules were bioequivalent in terms of AUC0-infinity but not in terms of Cmax; geometric-least-squares means ratios (with 90% confidence intervals) were 1.03 (0.92 to 1.14) and 1.25 (1.03 to 1. 53) for AUC0-infinity and Cmax, respectively. Administration of soft gelatin capsules of amprenavir with a high-fat breakfast resulted in a 14% decrease in the mean AUC0-infinity (from 9.58 to 8.26 microg. h/ml), which is not likely to be clinically significant. The most common adverse events related to amprenavir were headache, nausea, and hypesthesia. Amprenavir appears to be safe and well tolerated over the dose range of 150 to 1200 mg. On the basis of the present single-dose studies, amprenavir is an HIV protease inhibitor with favorable absorption and clearance pharmacokinetics that are only minimally affected by administration with food. PMID- 10390224 TI - Multiple mechanisms of action for inhibitors of histidine protein kinases from bacterial two-component systems. AB - Many pathogenic bacteria utilize two-component systems consisting of a histidine protein kinase (HPK) and a response regulator (RR) for signal transduction. During the search for novel inhibitors, several chemical series, including benzoxazines, benzimidazoles, bis-phenols, cyclohexenes, trityls, and salicylanilides, were identified that inhibited the purified HPK-RR pairs KinA Spo0F and NRII-NRI, with 50% inhibitory concentrations (IC50s) ranging from 1.9 to >500 microM and MICs ranging from 0.5 to >16 microg/ml for gram-positive bacteria. However, additional observations suggested that mechanisms other than HPK inhibition might contribute to antibacterial activity. In the present work, representative compounds from the six different series of inhibitors were analyzed for their effects on membrane integrity and macromolecular synthesis. At 4x MIC, 17 of 24 compounds compromised the integrity of the bacterial cell membrane within 10 min, as measured by uptake of propidium iodide. In this set, compounds with lower IC50s tended to cause greater membrane disruption. Eleven of 12 compounds inhibited cellular incorporation of radiolabeled thymidine and uridine >97% in 5 min and amino acids >80% in 15 min. The HPK inhibitor that allowed >25% precursor incorporation had no measurable MIC (>16 microg/ml). Fifteen of 24 compounds also caused hemolysis of equine erythrocytes. Thus, the antibacterial HPK inhibitors caused a rapid decrease in cellular incorporation of RNA, DNA, and protein precursors, possibly as a result of the concomitant disruption of the cytoplasmic membrane. Bacterial killing by these HPK inhibitors may therefore be due to multiple mechanisms, independent of HPK inhibition. PMID- 10390225 TI - Antifungal properties and target evaluation of three putative bacterial histidine kinase inhibitors. AB - Histidine protein kinases have been explored as potential antibacterial drug targets. The recent identification of two-component histidine kinases in fungi has led us to investigate the antifungal properties of three bacterial histidine kinase inhibitors (RWJ-49815, RWJ-49968, and RWJ-61907). All three compounds were found to inhibit growth of the Saccharomyces cerevisiae and Candida albicans strains, with MICs ranging from 1 to 20 microg/ml. However, deletion of SLN1, the only histidine kinase in S. cerevisiae, did not alter drug efficacy. In vitro kinase assays were performed by using the Sln1 histidine kinase purified from bacteria as a fusion protein to glutathione S-transferase. RWJ-49815 and RWJ 49968 inhibited kinase a 50% inhibitory concentration of 10 microM, whereas RWJ 61907 failed to inhibit at concentrations up to 100 microM. Based on these results, we conclude that these compounds have antifungal properties; however, their mode of action appears to be independent of histidine kinase inhibition. PMID- 10390226 TI - In vitro antifungal activity and cytotoxicity of a novel membrane-active peptide. AB - In the present study, we investigated the antifungal activity and cytotoxicity of a novel membrane-active peptide, KKVVFKVKFKK (MP). MP inhibited the growth of various pathogenic fungi isolated from patients and of fluconazole-resistant fungi at concentrations of 2 to 32 microg/ml. MP had potent fungicidal activity; the minimal fungicidal concentrations of the peptide were no more than fourfold greater than the MICs. Time course experiments of MP-induced killing of Candida albicans ATCC 36232 showed that the rate of killing was rapid and depended on the concentration of MP. MP had a strong synergism with other antifungal drugs; the fractional inhibitory concentration index values of MP with amphotericin B and fluconazole for C. albicans were 0.16 and 0.02, respectively. The 50% inhibitory concentrations of MP for NIH 3T3 and Jurkat cells were approximately 100 times higher than the MIC for C. albicans ATCC 36232, indicating that MP had high selectivity between the fungal and mammalian cells. These results suggest that MP has great advantages in the development of antifungal agents. PMID- 10390227 TI - Single-dose pharmacokinetics and safety of abacavir (1592U89), zidovudine, and lamivudine administered alone and in combination in adults with human immunodeficiency virus infection. AB - Abacavir (1592U89), a nucleoside reverse transcriptase inhibitor with in vitro activity against human immunodeficiency virus type-1 (HIV-1), has been evaluated for efficacy and safety in combination regimens with other nucleoside analogs, including zidovudine (ZDV) and lamivudine (3TC). To evaluate the potential pharmacokinetic interactions between these agents, 15 HIV-1-infected adults with a median CD4(+) cell count of 347 cells/mm3 (range, 238 to 570 cells/mm3) were enrolled in a randomized, seven-period crossover study. The pharmacokinetics and safety of single doses of abacavir (600 mg), ZDV (300 mg), and 3TC (150 mg) were evaluated when each drug was given alone or when any two or three drugs were given concurrently. The concentrations of all drugs in plasma and the concentrations of ZDV and its 5'-glucuronide metabolite, GZDV, in urine were measured for up to 24 h postdosing, and pharmacokinetic parameter values were calculated by noncompartmental methods. The maximum drug concentration (Cmax), the area under the concentration-time curve from time zero to infinity (AUC0 infinity), time to Cmax (Tmax), and apparent elimination half-life (t1/2) of abacavir in plasma were unaffected by coadministration with ZDV and/or 3TC. Coadministration of abacavir with ZDV (with or without 3TC) decreased the mean Cmax of ZDV by approximately 20% (from 1.5 to 1.2 microg/ml), delayed the median Tmax for ZDV by 0.5 h, increased the mean AUC0-infinity for GZDV by up to 40% (from 11.8 to 16.5 microg. h/ml), and delayed the median Tmax for GZDV by approximately 0.5 h. Coadministration of abacavir with 3TC (with or without ZDV) decreased the mean AUC0-infinity for 3TC by approximately 15% (from 5.1 to 4.3 microg. h/ml), decreased the mean Cmax by approximately 35% (from 1.4 to 0.9 microg/ml), and delayed the median Tmax by approximately 1 h. While these changes were statistically significant, they are similar to the effect of food intake (for ZDV) or affect an inactive metabolite (for GZDV) or are relatively minor (for 3TC) and are therefore not considered to be clinically significant. No significant differences were found in the urinary recoveries of ZDV or GZDV when ZDV was coadministered with abacavir. There was no pharmacokinetic interaction between ZDV and 3TC. Mild to moderate headache, nausea, lymphadenopathy, hematuria, musculoskeletal chest pain, neck stiffness, and fever were the most common adverse events reported by those who received abacavir. Coadministration of ZDV or 3TC with abacavir did not alter this adverse event profile. The three drug regimen was primarily associated with gastrointestinal events. In conclusion, no clinically significant pharmacokinetic interactions occurred between abacavir, ZDV, and 3TC in HIV-1-infected adults. Coadministration of abacavir with ZDV or 3TC produced mild changes in the absorption and possibly the urinary excretion characteristics of ZDV-GZDV and 3TC that were not considered to be clinically significant. Coadministration of abacavir with ZDV and/or 3TC was generally well tolerated and did not produce unexpected adverse events. PMID- 10390228 TI - Activities of sordarins in murine histoplasmosis. AB - Sordarins are new antifungals which inhibit fungal protein synthesis by blocking elongation factor 2. Three compounds were evaluated in a murine model of histoplasmosis. Immune-competent mice were infected intravenously with 10(6) to 10(8) CFU of Histoplasma capsulatum yeast cells. Mice were treated either orally with sordarins or fluconazole from day 2 through 8 after infection or intraperitoneally with amphotericin B during the same period. Protection was measured by increased rates of survival for 30 days after infection or reduction of lung or kidney tissue counts 9 days after infection. All three of the antifungal drugs tested were protective compared with controls. Sordarins were effective at doses as low as 2 mg/kg of body weight/day. This novel class of drugs compared favorably with amphotericin B and fluconazole for the treatment of histoplasmosis. PMID- 10390229 TI - Reversal of tetracycline resistance mediated by different bacterial tetracycline resistance determinants by an inhibitor of the Tet(B) antiport protein. AB - Active efflux is a useful strategy by which bacteria evade growth inhibition by antibiotics. Certain semisynthetic tetracycline (TC) analogs, substituted at the 13th carbon at C-6 on ring C of the TC molecule, blocked TC efflux as revealed in everted membrane vesicles from class B TC-resistant (Tcr) Escherichia coli (M. L. Nelson, B. H. Park, J. S. Andrews, V. A. Georgian, R. C. Thomas, and S. B. Levy, J. Med. Chem. 36:370-377, 1993). A representative C-13-substituted analog, 13 cyclopentylthio-5-OH-TC (13-CPTC), was shown to competitively inhibit TC translocation by the Tet(B) protein, blocking the uptake of TC into vesicles and therefore the efflux of TC from whole cells. Against Tcr E. coli, 13-CPTC, when used in combination with doxycycline, produced synergistic inhibition of growth. 13-CPTC was shown to increase the uptake of [3H]TC into the resistant cells. 13 CPTC alone was a potent growth inhibitor against TC-susceptible (Tcs) and Tcr Staphylococcus aureus and enterococci specifying class K or class L efflux dependent TC resistance mechanisms or, unexpectedly, the class M ribosomal protection mechanism. These findings indicate that derivatives of TC, identified by their ability to block the Tet(B) efflux protein, can restore TC activity against Tcr bacteria bearing either of the two known resistance mechanisms. Blocking drug efflux and increasing intracellular drug concentrations constitute an effective approach to reversing TC resistance and may be generally applicable to other antibiotics rendered ineffective by efflux proteins. PMID- 10390230 TI - Purification, reconstitution, and inhibition of cytochrome P-450 sterol delta22 desaturase from the pathogenic fungus Candida glabrata. AB - Sterol delta22-desaturase has been purified from a strain of Candida glabrata with a disruption in the gene encoding sterol 14alpha-demethylase (cytochrome P 45051; CYP51). The purified cytochrome P-450 exhibited sterol delta22-desaturase activity in a reconstituted system with NADPH-cytochrome P-450 reductase in dilaurylphosphatidylcholine, with the enzyme kinetic studies revealing a Km for ergosta-5,7-dienol of 12.5 microM and a Vmax of 0. 59 nmol of this substrate metabolized/min/nmol of P-450. This enzyme is encoded by CYP61 (ERG5) in Saccharomyces cerevisiae, and homologues have been shown in the Candida albicans and Schizosaccharomyces pombe genome projects. Ketoconazole, itraconazole, and fluconazole formed low-spin complexes with the ferric cytochrome and exhibited type II spectra, which are indicative of an interaction between the azole moiety and the cytochrome heme. The azole antifungal compounds inhibited reconstituted sterol delta22-desaturase activity by binding to the cytochrome with a one-to-one stoichiometry, with total inhibition of enzyme activity occurring when equimolar amounts of azole and cytochrome P-450 were added. These results reveal the potential for sterol delta22-desaturase to be an antifungal target and to contribute to the binding of drugs within the fungal cell. PMID- 10390231 TI - Effects of novel 6-desfluoroquinolones and classic quinolones on pentylenetetrazole-induced seizures in mice. AB - There have been several reports that convulsions, although rare, occur in patients who receive fluoroquinolones. In this study, the proconvulsant effects exhibited by a novel series of 6-desfluoroquinolones and some classic quinolones on pentylenetetrazole (PTZ)-induced seizures in mice were evaluated and compared. Animals were intraperitoneally injected with vehicle or quinolone derivatives (5 to 100 microg/g of body weight) 30 min before the subcutaneous (s.c.) administration of PTZ (40 microg/g). In each experiment, mice were then observed for 1 h to monitor for the incidence and onset of clonic seizures. The order of proconvulsant activity in our epileptic model was MF5184 > MF5187 > pefloxacin > MF5189 > ofloxacin > ciprofloxacin > MF5140 > MF5181 > MF5137 > rufloxacin > MF5143 > MF5158 > MF5191 > MF5128 > MF5138 > cinoxacin > MF5142 > norfloxacin > nalidixic acid. The relationship between the chemical structure and the proconvulsant activity of 6-desfluoroquinolone derivatives was studied. We observed that, in terms of toxicity to the central nervous system (CNS), besides the heterocyclic side chain (moiety) at the C-7 position, the C-6 substituent also appears to play an important role. In particular, a hydrogen at the C-6 position seemed to be responsible for major neurotoxic activity in comparison to an amino group located in the same position. The relationship between lipophilicity and proconvulsant activity was also investigated. We did not find any clear relationship between a higher level of lipophilicity and major proconvulsant properties. Although the principal mechanism by which quinolones induce potentiation of the proconvulsant effects of PTZ cannot be easily determined, it is possible that the convulsions are caused by drug interactions, because both PTZ and quinolones are believed to increase excitation of the CNS by inhibition of gamma-aminobutyric acid binding to receptors. PMID- 10390232 TI - Comparative efficacies of liposomal amikacin (MiKasome) plus oxacillin versus conventional amikacin plus oxacillin in experimental endocarditis induced by Staphylococcus aureus: microbiological and echocardiographic analyses. AB - Optimal treatment strategies for serious infections caused by Staphylococcus aureus have not been fully characterized. The combination of a beta-lactam plus an aminoglycoside can act synergistically against S. aureus in vitro and in vivo. MiKasome, a new liposome-encapsulated formulation of conventional amikacin, significantly prolongs serum half-life (t1/2) and increases the area under the concentration-time curve (AUC) compared to free amikacin. Microbiologic efficacy and left ventricular function, as assessed by echocardiography, were compared in animals administered either oxacillin alone or oxacillin in combination with conventional amikacin or MiKasome in a rabbit model of experimental endocarditis due to S. aureus. In vitro, oxacillin, combined with either free amikacin or MiKasome, prevented the bacterial regrowth observed with aminoglycosides alone at 24 h of incubation. Rabbits with S. aureus endocarditis were treated with either oxacillin alone (50 mg/kg, given intramuscularly three times daily), oxacillin plus daily amikacin (27 mg/kg, given intravenously twice daily), or oxacillin plus intermittent MiKasome (160 mg/kg, given intravenously, a single dose on days 1 and 4). The oxacillin-alone dosage represents a subtherapeutic regimen against the infecting strain in the endocarditis model (L. Hirano and A. S. Bayer, Antimicrob. Agents Chemother. 35:685-690, 1991), thus allowing recognition of any enhanced bactericidal effects between oxacillin and either aminoglycoside formulation. Treatment was administered for either 3 or 6 days, and animals were sacrificed after each of these time points or at 5 days after a 6-day treatment course (to evaluate for posttherapy relapse). Left ventricular function was analyzed by utilizing serial transthoracic echocardiography during treatment and posttherapy by measurement of left ventricular fractional shortening. At all sacrifice times, both combination regimens significantly reduced S. aureus vegetation counts versus control counts (P < 0.05). In contrast, oxacillin alone did not significantly reduce S. aureus vegetation counts after 3 days of therapy. Furthermore, at this time point, the two combinations were significantly more effective than oxacillin alone (P < 0.05). All three regimens were effective in significantly decreasing bacterial counts in the myocardium during and after therapy compared to controls (P < 0.05). In kidney and spleen abscesses, all regimens significantly reduced bacterial counts during therapy (P < 0.0001); however, only the combination regimens prevented bacteriologic relapse in these organs posttherapy. By echocardiographic analysis, both combination regimens yielded a significant physiological benefit by maintaining normal left ventricular function during treatment and posttherapy compared with oxacillin alone (P < 0.001). These results suggest that the use of intermittent MiKasome (similar to daily conventional amikacin) enhances the in vivo bactericidal effects of oxacillin in a severe S. aureus infection model and preserves selected physiological functions in target end organs. PMID- 10390233 TI - Class C beta-lactamases operate at the diffusion limit for turnover of their preferred cephalosporin substrates. AB - It has been suggested that class C beta-lactamases have evolved to carry out a metabolic reaction other than hydrolysis of beta-lactam antibiotics. It is demonstrated in the present study that the class C beta-lactamase from Enterobacter cloacae P99 has reached the diffusion limit in its ability to hydrolyze its preferred cephalosporin substrates. The increase in the solution viscosity by addition of a microviscogen (sucrose) caused the decline in the parameter kcat/Km for hydrolysis of cephaloridine and cephalosporin C (approximately 2.5-fold at a relative viscosity of 2.9). A similar increase in viscosity has no effect on the turnover rate of the poorer substrates cefepime and penicillin G. Addition of a macroviscogen (polyethylene glycol) to the reaction mixture did not change the rate of turnover for any of the substrates tested because in this case the viscogen would not interfere with the motion of small molecules, as was expected. Therefore, it would appear that the driving force behind the evolution of this class C beta-lactamase and, in principle, other enzymes of this class is indeed the functional reaction of this enzyme as a drug resistance factor. PMID- 10390234 TI - Combinations of vancomycin and beta-lactams are synergistic against staphylococci with reduced susceptibilities to vancomycin. AB - Evidence of synergism between combinations of vancomycin and beta-lactam antibiotics against 59 isolates of methicillin-resistant staphylococci (Staphylococcus aureus, Staphylococcus epidermidis, and Staphylococcus haemolyticus) for which vancomycin MICs ranged from 1 to 16 microg/ml were tested by broth microdilution checkerboard, disk diffusion, agar dilution, and time-kill antimicrobial susceptibility tests. The combination of vancomycin and oxacillin demonstrated synergy by all test methods against 30 of 59 isolates; no antagonism was seen. Synergy with vancomycin was also found by modified disk diffusion testing for ceftriaxone, ceftazidime, cefpodoxime, and amoxicillin-clavulanate but not for aztreonam. Evidence of synergy correlated directly with vancomycin MICs. The efficacy of vancomycin given alone and in combination with nafcillin was tested in the rabbit model of experimental endocarditis caused by three clinical isolates of glycopeptide-intermediate-susceptible S. aureus (GISA) (isolates HIP5827, HIP5836, and MU50). Two of the GISA isolates (isolates MU50 and HIP5836) were extremely virulent in this model, with 27 of 42 (64%) animals dying during the 3-day trial. Therapy with either vancomycin or nafcillin given as a single agent was ineffective for animals infected with HIP5827 or MU50. However, the combination of vancomycin and nafcillin resulted in a mean reduction of 4.52 log10 CFU/g of aortic valvular vegetations per g compared to the reduction for controls for animals infected with HIP5827 and a reduction of 4. 15 log10 CFU/g for animals infected with MU50. Renal abscesses caused by HIP5827 were sterilized significantly better with the combination of vancomycin and nafcillin than by either treatment alone. We conclude that the combination of vancomycin and beta-lactams with antistaphylococcal activity is an effective regimen for the treatment of infections with clinical strains of staphylococci which demonstrate reduced susceptibility to glycopeptides. PMID- 10390235 TI - Lysostaphin treatment of experimental aortic valve endocarditis caused by a Staphylococcus aureus isolate with reduced susceptibility to vancomycin. AB - The rabbit model of endocarditis was used to test the effectiveness of vancomycin and two different lysostaphin dosing regimens for the treatment of infections caused by a Staphylococcus aureus strain with reduced susceptibility to vancomycin (glycopeptide-intermediate susceptible S. aureus [GISA]). Vancomycin was ineffective, with no evidence of sterilization of aortic valve vegetations. However, rates of sterilization of aortic valve vegetations were significantly better for animals treated with either a single dose of lysostaphin (43%) or lysostaphin given twice daily for 3 days (83%) than for animals treated with vancomycin. Rabbits given a single dose of lysostaphin followed by a 3-day drug free period had mean reductions in aortic valve vegetation bacterial counts of 7.27 and 6.63 log10 CFU/g compared with those for untreated control rabbits and the vancomycin-treated group, respectively. We conclude that lysostaphin is an effective alternative for the treatment of experimental aortic valve endocarditis caused by a clinical VISA strain. PMID- 10390236 TI - Effect of fluoroquinolone concentration on selection of resistant mutants of Mycobacterium bovis BCG and Staphylococcus aureus. AB - When Mycobacterium bovis BCG and Staphylococcus aureus were plated on agar containing increasing concentrations of fluoroquinolone, colony numbers exhibited a sharp drop, followed by a plateau and a second sharp drop. The plateau region correlated with the presence of first-step resistant mutants. Mutants were not recovered at concentrations above those required for the second sharp drop, thereby defining a mutant prevention concentration (MPC). A C-8-methoxy group lowered the MPC for an N-1-cyclopropyl fluoroquinolone. PMID- 10390237 TI - Sequence of the MIR-1 beta-lactamase gene. AB - The complete nucleotide sequence of the plasmid-mediated MIR-1 beta-lactamase gene confirms its relationship to chromosomally located ampC genes of Enterobacter cloacae. blaMIR-1 is not part of a typical gene cassette but does lie near an element that could be involved in its capture on a plasmid. PMID- 10390238 TI - Characterization of new mutations in pyrazinamide-resistant strains of Mycobacterium tuberculosis and identification of conserved regions important for the catalytic activity of the pyrazinamidase PncA. AB - A new set of mutations, including transposition of the insertion sequence IS6110, was identified in the pncA gene from 19 pyrazinamide-resistant Mycobacterium tuberculosis strains. Alignment of the PncA protein from M. tuberculosis with homologous proteins from different bacterial species revealed three highly conserved regions in PncA which may play an important role in the processing of pyrazinamide. PMID- 10390240 TI - In vitro activity of HSR-903, a new oral quinolone, against bacteria causing respiratory infections. AB - The in vitro activity of HSR-903, an oral quinolone, against 196 recent clinical isolates of respiratory pathogens was evaluated. HSR-903 was 2 to 32 times more active than ofloxacin, ciprofloxacin, and sparfloxacin against Staphylococcus aureus, including methicillin-resistant strains, and Streptococcus pneumoniae and was at least as active as the other quinolones against gram-negative pathogens. PMID- 10390239 TI - pncA mutations in pyrazinamide-resistant Mycobacterium tuberculosis isolates from northwestern Russia. AB - Thirty-six pyrazinamide-resistant and eight pyrazinamide-susceptible Mycobacterium tuberculosis isolates from Russia were analyzed for their pncA mutations. Thirty-one (86.1%) of the resistant isolates had a mutation either in pncA or upstream of the gene. Twenty of the 23 different mutations found in this study had not been described earlier. pncA genotype correlated well with pyrazinamidase activity and BACTEC 460 susceptibility test results. PMID- 10390241 TI - Activation of the 2'-N-acetyltransferase gene [aac(2')-Ia] in Providencia stuartii by an interaction of AarP with the promoter region. AB - The aac(2')-Ia gene in Providencia stuartii encodes a 2'-N-acetyltransferase capable of acetylating both peptidoglycan and certain aminoglycoside antibiotics. Regulation of the aac(2')-Ia gene is influenced in a positive manner by the product of the aarP gene, which encodes a small transcriptional activator of the AraC (XylS) family. In this study, we demonstrate the sequence requirements at the aac(2')-Ia promoter for AarP binding and activation. PMID- 10390242 TI - In vitro activities of the everninomicin SCH 27899 and other newer antimicrobial agents against Borrelia burgdorferi. AB - The in vitro activity of the everninomicin antibiotic SCH 27899 against 17 isolates of Borrelia spp. was investigated. MICs ranged from 0.06 to 0.5 microg/ml. Time-kill studies with the B31 strain of B. burgdorferi demonstrated >/=3-log10-unit killing after 72 h with concentrations representing four times the MIC. The in vitro activity of four other newer antimicrobial agents, meropenem, cefepime, quinupristin-dalfopristin, and linezolid, was also tested against the B31 strain. Meropenem was the most potent of the latter agents, with an MIC of 0.125 microg/ml. PMID- 10390243 TI - Improvement of in vitro and in vivo antileishmanial activities of 2', 6' dihydroxy-4'-methoxychalcone by entrapment in poly(D,L-lactide) nanoparticles. AB - The inhibition of intracellular Leishmania amazonensis growth by 2', 6'-dihydroxy 4'-methoxychalcone (DMC) isolated from Piper aduncum was further enhanced after encapsulation of DMC in polymeric nanoparticles. Encapsulated DMC also showed increased antileishmanial activity in infected BALB/c mice, as evidenced by significantly smaller lesions and fewer parasites in the lesions. PMID- 10390244 TI - Rapid detection, by PCR and reverse hybridization, of mutations in the Helicobacter pylori 23S rRNA gene, associated with macrolide resistance. AB - A PCR-based reverse hybridization system (research prototype kit INNO-LiPA for H. pylori resistance) was developed and evaluated for simultaneous detection of 23S ribosomal DNA point mutations, associated with macrolide resistance in Helicobacter pylori. Fifty-seven H. pylori strains (51 natural, 6 laboratory derived artificial, 52 resistant, and 5 susceptible strains) were tested by PCR LiPA (detecting mutations A2115-->G, G2141-->A, A2142-->G, A2142-->C, A2143-->G, A2143-->C, and A2143-->T), DNA sequencing, restriction fragment length polymorphism, and/or hybridization to oligonucleotide probes. Results were highly concordant, but PCR-LiPA appears to be more sensitive for the simultaneous detection of multiple mutants. PMID- 10390245 TI - Anti-Toxoplasma gondii activities and structure-activity relationships of novel fluoroquinolones related to trovafloxacin. AB - Eleven novel fluoroquinolones closely related to trovafloxacin were evaluated for their in vitro activity against Toxoplasma gondii, and their structure-activity relationships were examined. The 50% inhibitory concentration (IC50) of trovafloxacin against T. gondii was 2.93 microM; the IC50 of the 11 analogs ranged from 0.53 to 14. 09 microM. Six analogs had IC50s lower than that of trovafloxacin. Examination of the structure-activity relationships of the compounds revealed that addition of a -CH3 at C-5 of the 1,8-naphthyridone ring, at C-2 of the azabicyclohexane ring, or on the -NH2 at the 6 position of the azabicyclohexane ring resulted in a four- to sixfold increase in activity. Moreover, replacement of 2,4-difluorophenyl by cyclopropyl at N-1 of the 1,8 naphthyridone ring increased activity twofold, and moving the -NH2 one atom further away from the azabicyclohexane ring decreased activity. There was no difference between the naphthyridone and quinolone analogs. These results indicate that structure-activity studies of compounds related to drugs active against T. gondii may be useful in producing compounds with more potent activities against the parasite. PMID- 10390246 TI - In vitro and in vivo activities of tea catechins against Helicobacter pylori. AB - The catechin epigallocatechin gallate showed the strongest activity of the six tea catechins tested against Helicobacter pylori (MIC for 50% of the strains tested, 8 microg/ml). It had bactericidal activity at pH 7 but not at pH /=0.3; 4) absence of patent ductus arteriosus, sepsis, major congenital malformation, congenital heart disease; and 5) no evidence of maternal HIV or hepatitis B infection. The dosage schedule was 0.25 mg/kg bid for 3 days, then 0.15 mg/kg bid for 3 days, then a 10% reduction in the dose every 3 days until a dose of 0.1 mg/kg had been given for 3 days, from which time a dose of 0.1 mg/kg qod was continued until 42 days after entry. The primary endpoint was the number of days on assisted ventilation after study entry. Secondary outcomes of interest included days on supplemental oxygen, days of hospitalization, and potential adverse effects, such as infection, gastrointestinal bleeding, left ventricular hypertrophy, and severe retinopathy of prematurity. RESULTS: Infants in the dexamethasone- and placebo-treated groups were similar in terms of baseline attributes, including birth weight, gestational age, gender, race, and ventilator settings at entry. Infants treated with dexamethasone were on assisted ventilation and supplemental oxygen for fewer days after study entry (median days on ventilator, 5th and 95th percentiles, 13 [1-64] vs 25 [6-104]; days on oxygen, 59 [6-247] vs 100 [11-346]). No differences were found in risk of death, infection, or severe retinopathy. In subgroup analyses, the association of dexamethasone with more rapid weaning from the ventilator was weaker among infants enrolled before the 16th day of life, infants with chest radiographs showing cystic changes and/or hyperinflation, and infants requiring an FIO2 >/=0.7 or a peak inspiratory pressure >/=19 at study entry. CONCLUSIONS: A 42-day tapering course of dexamethasone decreases the duration of ventilator and oxygen dependency in very low birth weight infants and is not associated with an increased risk of short-term adverse effects. PMID- 10390256 TI - Gatekeeping and referral of children and adolescents to specialty care. AB - OBJECTIVE: In this study we examined how gatekeeping arrangements influence referrals to specialty care for children and adolescents in private and Medicaid insurance plans. DESIGN/PARTICIPANTS: We conducted a prospective study of office visits (n = 27 104) made to 142 pediatricians in 94 practices distributed throughout 36 states in a national primary care practice-based research network. During 10 practice-days, physicians and patients completed questionnaires on referred patients, while office staff kept logs of all visits. Physicians used medical records to complete questionnaires for a subset of patients 3 months after their referral was made. RESULTS: Gatekeeping arrangements were common among children and adolescents with private (57.8%) and Medicaid (43.3%) insurance. Patients in gatekeeping plans were more likely to be referred with private (3. 16% vs 1.85% visits referred) and Medicaid (5.39% vs 3.73%) financing. Increased parental requests for specialty care among gatekeeping patients did not explain the increased referral rate. Physicians' reasons for making the referral were similar between the two groups. Physicians were less likely to schedule an appointment or communicate with the specialist for referred patients in gatekeeping plans. However, rates of physician awareness that a specialist visit occurred and specialist communication back to pediatricians did not differ between the two groups 3 months after the referrals were made. CONCLUSIONS: Gatekeeping arrangements are common among insured children and adolescents in the United States. Our study suggests that gatekeeping arrangements increase referrals from pediatricians' offices to specialty care and compromise some aspects of coordination. PMID- 10390257 TI - Utilization of physician offices by adolescents in the United States. AB - Recent guidelines for adolescent primary care call for the specification of clinical services by three adolescent age subgroups. Yet analyses of office visits have either merged adolescence into one stage or divided it at age 15 years. OBJECTIVE: To explore the utilization of physician offices in the United States by early (11-14 years), middle (15-17 years), and late (18-21 years) adolescents. DESIGN: Secondary analysis of the 1994 National Ambulatory Medical Care Survey, focusing on visits made by the three adolescent age groups. SETTING: Nationally representative sample of 2426 physicians in nonfederal, nonhospital offices. SUBJECTS: A total of 33 598 visits by patients of all ages, representing 681.5 million visits in 1994. MAIN OUTCOME MEASURES: Number of visits, health insurance, providers seen, duration of visits, reasons for visits, resulting diagnoses, and counseling provided. RESULTS: Adolescents aged 11 to 21 years made 9.1% (61.8 million) of the total office visits and represented 15.4% of the total US population in 1994. This underrepresentation in visits held across all three adolescent age subgroups. Within the adolescent cohort, whites were overrepresented relative to their population proportion (78.5% of visits, 67.6% of population) and blacks and Hispanic adolescents were underrepresented (8.3% and 9.3% of visits, 15.5% and 13.1% of population). Middle adolescence signaled a life turning point from male to female predominance in office visits. Peak lifetime uninsurance rates occurred at middle adolescence for females (18.7%) and late adolescence for males (24.0%). Between childhood and early adolescence, public insurance decreased from 24.7% to 15.7% and uninsurance increased from 12.7% to 19.7%. Pediatricians accounted for the highest proportion of early adolescent visits (41.2%), family physicians for middle adolescent visits (35.3%), obstetrician-gynecologists for late adolescent female visits (37.3%), and family physicians for late adolescent male visits (34.8%). Mean visit duration during adolescence was 16 minutes, did not differ by age subgroup or sex, exceeded that of children (14.6 minutes), and was shorter than that of adults (19.3 minutes). Obstetrician-gynecologists spent more time with adolescents than did other physicians. Education or counseling was included in 50.4% of adolescent visits, ranging from 65.1% for obstetrician-gynecologists to 34.8% for internists. During early adolescence, the leading reasons for both male and female visits were respiratory (19.4%), dermatological (10.0%), and musculoskeletal (9.7%). A similar profile was found for middle and late adolescent males. For middle and late adolescent females, the leading reason for visits was special obstetrical-gynecological examination (12.8% and 21.1%), and the leading diagnosis resulting from visits was pregnancy (9.5% and 20.4%). CONCLUSIONS: Adolescents underutilize physician offices and are more likely to be uninsured than any other age group. Visits are short, and counseling is not a uniform component of care. As adolescents mature, their providers, presenting problems, and resulting diagnoses change. The data from the National Ambulatory Medical Care Survey support a staged approach to adolescent preventive services, targeted to the needs of three age subgroups. PMID- 10390258 TI - Physically abused adolescents: behavior problems, functional impairment, and comparison of informants' reports. AB - OBJECTIVE: This study, like earlier studies that focused on younger abused children, ascertained whether physically abused adolescents exhibited increased internalizing and externalizing behaviors. Relevance to pediatric practice is discussed. DESIGN: A cross-sectional design was used to compare the behavior of physically abused adolescents and comparison adolescents using self-reports, parent reports, and teacher reports. The level of agreement among raters was also examined. PARTICIPANTS: The subjects were 99 physically abused adolescents between the ages of 12 and 18 years, who were recruited from Child Protective Services. Comparison subjects were 99 community-recruited nonabused adolescents who were matched for age, gender, and income with the abused adolescents. MEASURES: The behavior of the adolescents was assessed using the Child Behavior Checklist, and the comparable Youth Self-Report and Teacher Report Form, which are widely used measures of behavioral and emotional problems. The Child Global Assessment Scale was also used as a measure of functional impairment and of the need for mental health services. RESULTS: Parents and teachers rated the problems of abused adolescents as significantly greater than the problems of nonabused adolescents on all checklist subscales. Abused adolescents reported significantly greater problems only on externalizing behavior subscales. In addition, based on interviewer ratings, physically abused adolescents exhibited significantly greater functional impairment. CONCLUSIONS: Similar to previous research on abused children, physically abused adolescents exhibit externalizing and internalizing behavior problems and experience greater functional impairment. Parent, teacher, and adolescent reports of externalizing behaviors were similar, but physically abused adolescents reported fewer internalizing behaviors than did the other informants. PMID- 10390259 TI - Adapting the gang model: peer mentoring for violence prevention. AB - OBJECTIVES: This study assessed the effectiveness of an inner-city peer-mentoring program in modifying the attitudes and behaviors involving violence of preadolescent mentees. METHODS: In a case-matched cohort study involving 7- to 13 year-old children, 50 children enrolled in peer mentoring (case subjects) were compared with 75 control subjects. Case subjects were involved before enrollment in the community program in which the intervention occurred; control subjects lived in the same housing project and were matched with case subjects on age, sex, and census tract. A total of 19 community adolescents mentored the case subjects by designing and presenting violence prevention lessons. Two reliable self-report scales, Determining our Viewpoints of Violent Events and Normative Beliefs About Aggression Scale, were used to measure attitudinal change. Teachers completed the Revised Behavior Problem Checklist to assess changes in behavior. RESULTS: At baseline, the survey scores of the case and control subjects were not different. After the intervention period, the case scores indicated less support for violence than the control scores. Case behavior scores did not change, but control behavior scores worsened. CONCLUSIONS: The data suggest that peer mentoring for younger children may be an important component of efforts to reduce youth violence. A larger multisite trial is warranted. PMID- 10390260 TI - Symptomatic congenital cytomegalovirus infection in infants born to mothers with preexisting immunity to cytomegalovirus. AB - OBJECTIVES: To determine the frequency of symptomatic congenital cytomegalovirus (CMV) infection in the offspring of women with a recurrent maternal CMV infection and to characterize the demographic and newborn findings. METHODS: Study subjects consisted of infants with symptomatic congenital CMV infection identified by a newborn virologic screening program at the University of Alabama Hospital between 1991 and 1997 and were enrolled in a long-term follow-up study. Maternal infections were categorized by an analysis of archival serum specimens collected before pregnancy and at the time of delivery. Demographic data and clinical findings at birth were collected from maternal and newborn hospital records and from parents at the time of initial evaluation. RESULTS: Of the 47 infants with symptomatic congenital CMV infection identified during the study period, 8 were born to mothers with a confirmed nonprimary or recurrent CMV infection. The type of maternal infection could be ascertained in only approximately 43% (20/47) of the children with symptomatic congenital CMV infection born at the University of Alabama Hospital during the study period. There were no significant differences in demographic characteristics of the recurrent infection group and the infants who were born to mothers with either primary CMV infection during pregnancy or unclassified maternal infection. Similarly, the range of severity of clinical abnormalities during the newborn period did not differ in the two groups of children. Furthermore, there were no significant differences in the incidence of sequelae at long-term follow-up in the two groups of children. CONCLUSIONS: Symptomatic congenital CMV infection can occur after a nonprimary or recurrent maternal infection. However, the exact incidence of symptomatic congenital CMV infection among children born to women with preexisting immunity remains to be defined. PMID- 10390261 TI - Evidence for independent genetic influences on fat mass and body mass index in a pediatric twin sample. AB - OBJECTIVE: Insight into genetic and environmental influences on fat mass, independent of body mass index (BMI; kg/m2), is expected to enhance methods for treating pediatric obesity. However, few studies have estimated the heritability of fat mass in pediatric samples, and those conducted have relied primarily on BMI measurements. PRESENT STUDY: Using bioimpedance analysis, the present study tested a series of hypotheses predicting significant genetic and environmental influences on percent body fat (PBF) above and beyond BMI. Subjects were 66 pairs of twins, including 41 monozygotic and 25 dizygotic pairs, from 3 to 17 years of age. Structural equation modeling tested hypotheses, adjusting for demographic variables. RESULTS: Analyses indicated significant genetic influences on PBF, with genes estimated to account for 75% to 80% of the phenotypic variation. The remaining variation was attributable to nonshared environmental influences. Multivariate analyses revealed sizable genetic correlations and environmental correlations between BMI and PBF (rg =.74 and re =.67, respectively), suggesting that some genes and environmental experiences influence both phenotypes. However, analyses confirmed genetic and environmental influences on PBF above and beyond BMI. For example, 62.5% of the total genetic variation in PBF was attributable to genes that influenced PBF but not BMI. CONCLUSION: There seems to be a substantial genetic contribution to fat mass distinct from BMI in a sample of children and adolescents. Studies testing putative genetic or environmental determinants of pediatric obesity might be strengthened further by including research-based body composition methods. PMID- 10390262 TI - Neonatal hyperbilirubinemia in glucose-6-phosphate dehydrogenase-deficient heterozygotes. AB - OBJECTIVES: We assessed the incidence of hyperbilirubinemia, defined as serum total bilirubin >/=15 mg/dL (256 micromol/L), in a cohort of Sephardic Jewish female neonates at risk for glucose-6-phosphate dehydrogenase (G-6-PD) deficiency with especial emphasis on the heterozygotes. We studied the roles of hemolysis by blood carboxyhemoglobin (COHb) determinations and of the variant promoter of the gene for the bilirubin-conjugating enzyme uridine 5'-diphosphate glucuronosyltransferase 1 (UGT1A1) seen in Gilbert's syndrome in the pathogenesis of the hyperbilirubinemia. METHODS: Consecutively born, healthy, term, female neonates were screened for G-6-PD deficiency and observed clinically with serum bilirubin evaluations as indicated for hyperbilirubinemia. On day 3, blood was sampled for COHb, total hemoglobin (tHb), and a mandatory serum bilirubin determination. COHb, determined by gas chromatography, was expressed as percentage of tHb and corrected for inspired carbon monoxide (COHbc). DNA was analyzed for the G-6-PD Mediterranean563T mutation and for the variant UGT1A1 gene. RESULTS: The cohort included 54 G-6-PD-deficient heterozygotes, 19 deficient homozygotes, and 112 normal homozygotes. More heterozygotes (12/54, 22%; relative risk: 2.26; 95% CI: 1.07-4.80) and deficient homozygotes (5/19, 26.3%; relative risk: 2.68; 95% CI: 1.05-6.90) developed hyperbilirubinemia, than did normal homozygotes (11/112, 9.8%). Third-day serum bilirubin values that were obtained from 144 neonates were significantly higher in both heterozygotes (11.2 +/- 3. 7 mg/dL [192 +/- 64 micromol/L]) and G-6-PD-deficient homozygotes (12.0 +/ 3.0 mg/dL [206 +/- 52 micromol/L]) than in the G-6-PD normal homozygotes (9.4 +/ 3.4 mg/dL [160 +/- 58 micromol/L). In contrast, COHbc values were higher only in G-6-PD-deficient homozygotes (0.74% +/- 0.14%) and not in heterozygotes (0.69% +/ 0. 19%, not statistically significant), compared with control values (0. 63% +/- 0.19%). High COHbc values were not a prerequisite for the development of hyperbilirubinemia in any of the G-6-PD genotypes. A greater incidence of hyperbilirubinemia was found among the G-6-PD-deficient heterozygotes, who also had the variant UGT1A1 gene, in both heterozygous (6/20, 30%) and homozygous (4/8, 50%) forms, than was found in their counterparts with the normal UGT1A1 gene (2/26, 7.7%). This effect was not seen in the G-6-PD normal homozygote group. A color reduction screening test for G-6-PD deficiency identified only 20.4% (11/54) of the heterozygotes. CONCLUSIONS: We showed that G-6-PD-deficient heterozygotes, categorically defined by DNA analysis, are at increased risk for neonatal hyperbilirubinemia. The screening test that was used was unable to detect most heterozygotes. Increased bilirubin production was not crucial to the development of hyperbilirubinemia, but presence of the variant UGT1A1 gene did confer increased risk. PMID- 10390263 TI - Comparative analysis of serologic screening tests for the initial diagnosis of celiac disease. AB - OBJECTIVE: To prospectively evaluate and compare the sensitivity, specificity, and positive and negative predictive values of serum antigliadin (AGA) and antiendomysium antibodies (EMA) in predicting the initial diagnosis of celiac disease. DESIGN: Sera were tested prospectively for IgA and IgG AGA by enzymed linked immunosorbent assay and IgA EMA by immunofluorescence techniques on monkey esophagus and human umbilical cord sections in 95 pediatric patients referred for duodenal biopsies. PATIENTS: Ninety-five pediatric patients were referred for duodenal biopsies, with a clinical suspicion of celiac disease; 24 of those patients had celiac disease by criteria of the European Society for Pediatric Gastroenterology and Nutrition. SETTING: A pediatric gastroenterology clinic of a tertiary care pediatric university hospital. RESULTS: EMA testing on human umbilical cords was the most specific but was also the least sensitive. All the patients with biopsy-proven celiac disease were identified by either one or both serologic tests (100% combined sensitivity). The combination of AGA and EMA on monkey esophagus resulted in a negative predictive value of 100% accuracy. CONCLUSIONS: A combination of AGA and EMA tests resulted in 100% sensitivity and 100% negative predictive value, useful in selecting patients for duodenal biopsy. PMID- 10390264 TI - Oral versus initial intravenous therapy for urinary tract infections in young febrile children. AB - BACKGROUND: The standard recommendation for treatment of young, febrile children with urinary tract infection has been hospitalization for intravenous antimicrobials. The availability of potent, oral, third-generation cephalosporins as well as interest in cost containment and avoidance of nosocomial risks prompted evaluation of the safety and efficacy of outpatient therapy. METHODS: In a multicenter, randomized clinical trial, we evaluated the efficacy of oral versus initial intravenous therapy in 306 children 1 to 24 months old with fever and urinary tract infection, in terms of short-term clinical outcomes (sterilization of the urine and defervescence) and long-term morbidity (incidence of reinfection and incidence and extent of renal scarring documented at 6 months by 99mTc-dimercaptosuccinic acid renal scans). Children received either oral cefixime for 14 days (double dose on day 1) or initial intravenous cefotaxime for 3 days followed by oral cefixime for 11 days. RESULTS: Treatment groups were comparable regarding demographic, clinical, and laboratory characteristics. Bacteremia was present in 3.4% of children treated orally and 5.3% of children treated intravenously. Of the short-term outcomes, 1) repeat urine cultures were sterile within 24 hours in all children, and 2) mean time to defervescence was 25 and 24 hours for children treated orally and intravenously, respectively. Of the long-term outcomes, 1) symptomatic reinfections occurred in 4.6% of children treated orally and 7.2% of children treated intravenously, 2) renal scarring at 6 months was noted in 9.8% children treated orally versus 7.2% of children treated intravenously, and 3) mean extent of scarring was approximately 8% in both treatment groups. Mean costs were at least twofold higher for children treated intravenously ($3577 vs $1473) compared with those treated orally. CONCLUSIONS: Oral cefixime can be recommended as a safe and effective treatment for children with fever and urinary tract infection. Use of cefixime will result in substantial reductions of health care expenditures. PMID- 10390265 TI - Prevalence and parental origin in Tetralogy of Fallot associated with chromosome 22q11 microdeletion. AB - OBJECTIVE: Tetralogy of Fallot is a common cardiac anomaly that is associated with chromosome 22q11 microdeletion. In this study we examined the mode of transmission as well as the parental origin of microdeletion in patients with tetralogy of Fallot. METHODS: Eighty-four children with sporadic tetralogy of Fallot (40 boys and 44 girls; mean age, 34 months) were analyzed for microdeletion at chromosome 22q11 by genotype analysis, using five microsatellite markers, D22S427, D22S941, D22S944, D22S264 and D22S311, and confirmed by quantitative polymerase chain reaction, using TUPLE1 and D22S264. All parents of these subjects consented to their own participation and their child's participation in the clinical evaluation and molecular study. To provide a molecular characterization of microdeletion, we isolated DNA from the parents and typed their DNA with each of the five polymorphic markers. RESULTS: Sixty-six patients were associated with pulmonary stenosis; and 8 of these cases (12%) had microdeletion. Eighteen patients were associated with pulmonary atresia, and 6 (33%) of these cases had microdeletion. The parental origins of the 14 patients with microdeletion were paternal in 3 cases and maternal in 11 cases. The most common mode of transmission was de novo without parental hemizygosity (93%). Transmission by autosomal dominant heredity was uncommon (7%). CONCLUSIONS: Biased parental origin was consistently found in tetralogy of Fallot patients with chromosomal 22q11 microdeletion. Our results indicated a higher prevalence of microdeletion because of inheritance of maternal microdeletion (78%). PMID- 10390266 TI - A three-day course of dexamethasone therapy to prevent chronic lung disease in ventilated neonates: a randomized trial. AB - BACKGROUND: Although several trials of early dexamethasone therapy have been completed to determine if such therapy would reduce mortality and chronic lung disease (CLD) in infants with respiratory distress, optimal duration and side effects of such therapy remain unknown. PURPOSE: The purpose of this study was: 1) to determine if a 3-day course of early dexamethasone therapy would reduce CLD and increase survival without CLD in neonates who received surfactant therapy for respiratory distress syndrome and 2) to determine adverse effects associated with such therapy. DESIGN: This was a prospective multicenter randomized trial comparing a 3-day course of dexamethasone therapy beginning at 24 to 48 hours of life to placebo therapy. Two hundred forty-one neonates (dexamethasone n = 118, placebo n = 123), who weighed between 500 g and 1500 g, received surfactant therapy, and were at significant risk for CLD or death using a model to predict CLD or death at 24 hours of life, were enrolled in the trial. Infants randomized to receive early dexamethasone were given 6 doses of dexamethasone at 12-hour intervals beginning at 24 to 48 hours of life. The primary outcomes compared were survival without CLD and CLD. CLD was defined by the need for supplemental oxygen at the gestational age of 36 weeks. Complication rates and adverse effects of study drug therapy were also compared. RESULTS: Neonates randomized to early dexamethasone treatment were more likely to survive without CLD (RR: 1.3; 95% CI: 1.03, 1.7) and were less likely to develop CLD (RR: 0.6; CI: 0.3, 0. 98). Mortality rates were not significantly different. Subsequent dexamethasone therapy use was less in early dexamethasone-treated neonates (RR: 0.8; CI: 0.7, 0.96). Very early (-2 SD) supplement treated children than in normal-weight children on placebo (9.8 vs 4.4 per 10(3) child-weeks; rate ratio: 2.21 [95% CI: 1.24-3.93]). By logistic regression analysis the risk of ALRI was lower in underweight supplement-treated children than in underweight children on placebo (point estimate 0.148 [95% CI: 0.034 0.634]). In contrast, risk of ALRI was higher in normal-weight supplement-treated children (WAZ >-1 SD to mean) than in normal-weight children on placebo in the same WAZ stratum (point estimate: 2.51 [95% CI: 1.24-5.05]). The risk of severe diarrhea was lower in supplement-treated children 18 to 23 months of age than in children on placebo in this age group (point estimate: 0.26 [95% CI: 0.06-1.00]). CONCLUSIONS: Weekly low-dose (10 000 IU) vitamin A supplementation in a region of subclinical deficiency protected underweight children from ALRI and paradoxically increased ALRI in normal children with body weight over -1 SD. Protection from severe diarrhea was consistent with previous trials. Additional research is warranted to delineate potential beneficial and detrimental interactions between nutritional status and vitamin A supplementation regarding ALRI. PMID- 10390288 TI - Rehospitalization in the first two weeks after discharge from the neonatal intensive care unit. AB - BACKGROUND: High-risk newborns are known to have higher than average utilization of services after discharge from the neonatal intensive care unit (NICU). Most studies on this subject report aggregate data over periods ranging from 1 to 3 years postdischarge. Little is known about events that are temporally close to NICU discharge. OBJECTIVES: To characterize rehospitalizations within the first 2 weeks after discharge from six community NICUs. METHODS: We scanned electronic databases and reviewed the charts of rehospitalized infants from six NICUs in the Kaiser Permanente Medical Care Program. We subdivided infants into five groups based on gestational age (GA) and birth hospitalization length of stay (LOS): 1) >/=37 weeks' GA with <4 days LOS (n = 2593); 2) >/=37 weeks' GA with >/=4 days' LOS (n = 1133); 3) from 33 to 36 weeks' GA with <4 days' LOS (n = 545); 4) from 33 to 36 weeks' GA with >/=4 days' LOS (n = 1196); and 5) <33 weeks' GA (n = 587). We performed bivariate and multivariate analyses to identify predictors that might be useful for practitioners. RESULTS: There were 6054 newborns discharged alive from the six study NICUs between August 1, 1992 and December 31, 1995, and 99.5% of these infants remained in the health plan during the 2 weeks after NICU discharge. The overall rehospitalization rate was 2.72%, which is 20% higher than the rate among healthy term newborns in the Kaiser Permanente Medical Care Program (2.26%). The two most common reasons for rehospitalization were jaundice (62/165, 37.6%) and feeding difficulties (25/165, 15.2%). Infants with 33 to 36 weeks' GA and <4 days' LOS were rehospitalized at a significantly higher rate than were all other infants (5.69%); 71% of infants in this group were rehospitalized for jaundice. The following variables predicted rehospitalization in multivariate models: <33 weeks' GA (adjusted OR [AOR]: 1.88; 95% CI: 1.10 3.21), from 33 to 36 weeks' GA with <96 hours' LOS (AOR: 2.94; 95% CI: 1.87 4.62), and birth at facility B, which had the highest rehospitalization rate of the six facilities (AOR: 1.92; 95% CI: 1.39-2.65). CONCLUSIONS: The rate of rehospitalization among NICU graduates is higher than among healthy term infants. Most of the rehospitalizations among infants with from 33 to 36 weeks' GA and <4 days' LOS are for illnesses that are not life-threatening. Collaborative studies and new process and outcomes measures are needed to assess the effectiveness of follow-up strategies in high-risk newborns. PMID- 10390289 TI - Early initiation of breastfeeding and the risk of infant diarrhea in rural Egypt. AB - BACKGROUND: Initiation of breastfeeding shortly after delivery may enhance breastfeeding's protective effect against diarrhea because of the protective properties of human colostrum contained in early breast milk. OBJECTIVE: To evaluate whether initiation of breastfeeding within the first 3 days of life, when breast milk contains colostrum, was associated with a lower rate of diarrhea in rural Egyptian infants during the first 6 months of life. METHODS: Infants initially breastfed (n = 198) were monitored prospectively with twice-weekly home visits to ascertain dietary practices and diarrheal illnesses. RESULTS: The burden of diarrhea during the first 6 months of life in the cohort was high: seven episodes per child-year of follow-up. Only 151 (76%) infants initiated breastfeeding during the first 3 days of life ("early initiation"). Infants in whom breastfeeding was initiated early had a 26% (95% CI: 2%,44%) lower rate of diarrhea than those initiated late. The protective association between early initiation and diarrhea was independent of the pattern of postinitiation dietary practices and was evident throughout the first 6 months of life. CONCLUSIONS: Early initiation of breastfeeding was associated with a marked reduction of the rate of diarrhea throughout the first 6 months of life, possibly because of the salutary effects of human colostrum. These data highlight the need for interventions to encourage early initiation of breastfeeding in less developed settings. PMID- 10390291 TI - The effects of different resistance training protocols on muscular strength and endurance development in children. AB - BACKGROUND: Previous research has shown that children can increase their muscular strength and muscular endurance as a result of regular participation in a progressive resistance training program. However, the most effective exercise prescription regarding the number of repetitions remains questionable. OBJECTIVE: To compare the effects of a low repetition-heavy load resistance training program and a high repetition-moderate load resistance training program on the development of muscular strength and muscular endurance in children. Design. Prospective, controlled trial. SETTING: Community-based youth fitness center. SUBJECTS: Eleven girls and 32 boys between the ages of 5.2 and 11.8 years. INTERVENTION: In twice-weekly sessions of resistance training for 8 weeks, children performed 1 set of 6 to 8 repetitions with a heavy load (n = 15) or 1 set of 13 to 15 repetitions with a moderate load (n = 16) on child-size exercise machines. Children in the control group (n = 12) did not resistance train. One repetition maximum (RM) strength and muscular endurance (repetitions performed posttraining with the pretraining 1-RM load) were determined on the leg extension and chest press exercises. RESULTS: One RM leg extension strength significantly increased in both exercise groups compared with that in the control subjects. Increases of 31.0% and 40.9%, respectively, for the low repetition-heavy load and high repetition-moderate load groups were observed. Leg extension muscular endurance significantly increased in both exercise groups compared with that in the control subjects, although gains resulting from high repetition-moderate load training (13.1 +/- 6.2 repetitions) were significantly greater than those resulting from low repetition-heavy load training (8.7 +/- 2.9 repetitions). On the chest press exercise, only the high repetition-moderate load exercise group made gains in 1-RM strength (16.3%) and muscular endurance (5.2 +/- 3.6 repetitions) that were significantly greater than gains in the control subjects. CONCLUSION: These findings support the concept that muscular strength and muscular endurance can be improved during the childhood years and favor the prescription of higher repetition-moderate load resistance training programs during the initial adaptation period. PMID- 10390290 TI - Pituitary gland gumma in congenital syphilis after failed maternal treatment: a case report. AB - A preterm, very low birth weight infant was born to a mother with early latent syphilis who was treated 10 days and 3 days before delivery with 2.4 mU of benzathine penicillin. The infant had clinical, laboratory, and radiographic abnormalities consistent with congenital syphilis, ie, a Venereal Disease Research Laboratory test titer that was fourfold greater than was the maternal titer, hepatosplenomegaly, abnormal liver function tests, pneumonitis, osteochondritis of the long bones, and cerebrospinal fluid (CSF) examination showing a reactive Venereal Disease Research Laboratory test, pleocytosis, and elevated protein content. The infant died on the third day of life, and an autopsy revealed an evolving gumma of the anterior pituitary. Immunoglobulin M immunoblotting of serum and CSF was positive, and polymerase chain reaction detected Treponema pallidum DNA in endotracheal aspirate and CSF. This case highlights the pathologic abnormalities observed in congenital syphilis and focuses on the rare finding of an evolving anterior pituitary gumma. Furthermore, it documents the failure of maternal syphilis treatment during the last 4 weeks of pregnancy to cure fetal infection and supports the recommendation that all infants born to mothers with syphilis treated during the last 4 weeks of pregnancy should receive penicillin therapy. PMID- 10390292 TI - Unexpected non-HIV causes of death in children born to HIV-infected mothers. Pediatric Pulmonary and Cardiac Complications of Vertically Transmitted HIV Infection Study Group. AB - INTRODUCTION: A high incidence of sudden, unexplained deaths in infants born to HIV-infected mothers has been noted in several epidemiologic studies. During the course of a prospective study of heart and lung disease in children born to HIV infected mothers, we noted that of 5 unexpected non-HIV-related deaths, 4 were attributed to traumatic events. METHODS: The Pediatric Pulmonary and Cardiac Complications of Vertically Transmitted HIV Infection (P2C2) study is a multicenter, prospective investigation of the incidence of heart and lung disease in HIV-infected children. A total of 805 children were enrolled and followed for 5 to 7 years with serial immunologic, pulmonary and cardiac function studies. During the study, a multidisciplinary committee was formed to review the cause of death for those patients who died. The committee used results of pulmonary, cardiac, and laboratory tests, hospital summaries, as well as autopsy and coroners' reports. The committee formed a consensus about the underlying and contributing causes of death for each subject using the definitions from the 1989 US Standard Certificate of Death. RESULTS: A total of 121 deaths occurred during the course of the P2C2 study. Of the 121 deaths, 5 were traumatic or sudden and unexpected and judged by the Mortality Review Committee to be unrelated to HIV infection. The median age at the time of death was 1.3 months and ranged from 1.2 to 37.8 months. Two infants died of trauma: a skull fracture and subdural hematoma in 1 infant and multiple skeletal fractures consistent with battered child syndrome in the other infant. The third infant died of accidental suffocation at home at 1.2 months of age. The fourth infant died suddenly and unexpectedly at home at 1.3 months of age. The autopsy showed no sign of HIV or other infection and was consistent with sudden unexpected death or SIDS. One non HIV-related death occurred when a 38-month-old child died together with the mother in an unwitnessed drowning. The cumulative mortality rate attributable to trauma and sudden death at 4 months of age was 0.95% (95% CI: 0.02-1. 87%) and the infant mortality rate was 9.5/1000 live births. Three children were born prematurely at 30, 33, and 36 weeks' gestational age, respectively, and 3 mothers admitted using recreational drugs before or during pregnancy. DISCUSSION: These traumatic and sudden non-HIV-related deaths accounted for 4.1% (5/121) of the deaths during the entire P2C2 study period and for 20% (4/20) of the deaths in the first year of life. Four deaths were attributable to accidental and nonaccidental trauma rather than to other common causes of infant death. One death was a sudden unexpected death, similar to SIDS, a leading cause of infant death in the United States. The majority of previously reported non-HIV-related deaths in infants born to HIV-infected mothers have been attributed to SIDS or to unexplained sudden death. In contrast with other reports, 4 of the 5 children in our series died of accidental or nonaccidental trauma and only 1 was a sudden unexplained death. It is unlikely that HIV exposure is related directly to the deaths described in this report; however, maternal HIV infection may be a marker for factors that place the child at risk for sudden or traumatic death. SUMMARY: This report suggests that children born to HIV-infected mothers may be at increased risk for traumatic or sudden, unexplained, non-HIV-related death. These children seem to be at risk regardless of their own HIV infection status. Furthermore, 4 of the deaths in our study occurred within the first few months of life, suggesting that this is a period of increased vulnerability. Studies to identify associated risk factors for non-HIV-related deaths are needed to identify these high-risk infants. Children born to HIV-infected mothers may be more vulnerable than was recognized previously and may be in need of increased social services, especially in early infancy. PMID- 10390293 TI - Rollover injuries in residential driveways: age-related patterns of injury. AB - BACKGROUND: The major objective of the present study was to determine the severity of nonfatal injuries sustained by children (<16 years old) when a motor vehicle rolls over them. We also sought to determine whether younger children (<24 months old) demonstrated different patterns of injury and/or a worse outcome, compared with older children (>24 months old). METHODS: We reviewed the medical records of 3971 consecutive admissions to a single trauma service at an urban children's hospital between March 1990 and October 1994. During this time period, 26 (0.7%) children presented with rollover injuries incurred by motor vehicles in residential driveways. Outcome was measured by length of both intensive care unit admission and hospitalization. RESULTS: Two children died shortly after admission and were excluded from the remainder of the study. Younger children (<24 months old) had significantly higher injury severity scores and lower pediatric trauma scale scores. Both the duration in the intensive care unit and the length of hospitalization were significantly longer in younger children, compared with children >24 months old. One explanation for these observations was that younger children had a significantly higher incidence of both head and neck and extremity injury but a similar incidence and severity of chest and abdominal trauma, compared with older children. Injuries requiring operative intervention were rare. CONCLUSION: Younger patients sustaining rollover injuries in the residential driveway have a worse outcome, in part, because of the head and neck or extremity injures that they incur. The majority of rollover injuries can be managed conservatively. pediatric trauma, driveway, pedestrian events, rollover injuries, injury severity score, pediatric trauma scale. PMID- 10390294 TI - Immigration and tuberculosis among children on the United States-Mexico border, County of San Diego, California. AB - OBJECTIVE: To identify factors contributing to a 400% increase in tuberculosis among children in San Diego County, California, from 1985 to 1993. DESIGN: Review of medical records of reported cases in 1989, 1991, and 1993 and their source case. RESULTS: Of 192 children with tuberculosis, the largest increase was observed in children younger than 5 years old, of whom 77.4% were born in the United States, 67.8% had a foreign-born parent, 73.1% came from a non-English speaking household, and 46.2% were known to visit Mexico. Of 28 source cases, 82.1% were born outside the United States, primarily in Mexico (67.9%). Resistance to at least one first-line antituberculous drug was identified in 27.5% of isolates from children and in 33.3% of isolates from source cases. CONCLUSIONS: The increase in tuberculosis and high level of drug-resistance among children born in the United States may be attributed to transmission outside of the United States or within the United States from household contacts born in countries in which tuberculosis is highly endemic. PMID- 10390295 TI - Pediatric myocardial infarction after racemic epinephrine administration. AB - Myocardial infarction is a previously unreported complication of treatment with racemic epinephrine that is used commonly in the emergency department for severe respiratory distress in bronchiolitis or croup syndrome. We describe a pediatric patient who presented with the croup syndrome and severe respiratory distress that required multiple doses of nebulized racemic epinephrine in the emergency department. The patient developed ventricular tachycardia and mild chest discomfort during one treatment, which resolved spontaneously on discontinuation of the nebulization. Persistently abnormal electrocardiograms and elevated creatine phosphokinase MB isoenzyme (CPK-MB) levels suggested a myocardial infarction had occurred. Subsequent echocardiography, cardiac catheterization, and angiography revealed an anatomically normal heart with normal coronary circulation; however, a stress nuclear study showed a small myocardial infarct. The significance of this previously unreported complication of racemic epinephrine is discussed, along with recommendations for proper use in the emergency department. PMID- 10390296 TI - Kawasaki disease: more patients are being diagnosed who do not meet American Heart Association criteria. AB - OBJECTIVE: To determine the frequency of Kawasaki disease (KD) diagnosis in patients who did and did not meet American Heart Association (AHA) diagnostic criteria and to examine the clinical findings, the time to treatment, and the outcomes of the two groups. DESIGN: Retrospective review of all patients with a discharge diagnosis of KD at a tertiary care children's hospital (1991-1997). RESULTS: A total of 127 patients were identified. All received intravenous immune globulin (IVIG) and had complete echocardiographic studies. AHA criteria were met in 81 (63.8%). More patients who did not meet criteria (9 of 46, 20%) had coronary artery abnormalities (CAA), compared with those who had the complete clinical picture (6 of 81, 7%). The 15 patients with CAA received IVIG later (12.4 +/- 7.4 days) from onset of symptoms compared with those with no CAA (8.2 +/- 4.6). The time period was the same for patients with CAA who met the criteria, (11.8 +/- 5.8 days) as for patients who did not meet AHA criteria (12.8 +/- 8.6 days). Infants were more likely than were older children to develop CAA, to receive IVIG later, and to be diagnosed with an incomplete clinical picture. CONCLUSION: Physicians are increasingly likely to diagnose KD in patients who do not meet complete AHA criteria. Despite the potential risks of overdiagnosis and overtreatment, this practice seems justified because the complete criteria are an insensitive indicator of having or developing CAA. PMID- 10390297 TI - Serum concentrations of antidepressants and benzodiazepines in nursing infants: A case series. AB - OBJECTIVE: The relative risk of psychotropic medication use in women with puerperal psychiatric illness who are breastfeeding has yet to be quantified adequately. Although the emotional and medical benefits of breastfeeding and adverse effects of maternal depression on infant development are well described, how these absolute benefits weigh against the potential effects of psychotropic drug use during lactation to ultimately guide clinical decisions is still unclear. The objective of this report was to evaluate the extent that psychotropic medications were present in the serum of infants breastfed by mothers treated with antidepressants and benzodiazepines. DESIGN: Serum samples were obtained from 35 nursing infants whose mothers were treated with psychotropic medications while breastfeeding. When a detectable concentration of medication was reported, information regarding infant behavior was obtained by maternal report. SETTING: The Perinatal and Reproductive Psychiatry Program at Massachusetts General Hospital serves as a regional consultation center for the treatment of psychiatric disorders during pregnancy and the postpartum period. PATIENTS: Subjects were mothers referred to the Perinatal Psychiatry Program for consultation regarding the relative safety of psychotropic medication use while breastfeeding. PRIMARY OUTCOME MEASURES: Presence of detectable levels of medication in infants whose mothers breastfed while taking psychotropic medications during pregnancy and/or during the puerperium and the well-being (based on maternal report) of infants who had detectable serum concentrations of medication. RESULTS: Seventy-four percent (n = 26) of infants had serum medication concentrations below the laboratory limit of detection (assay sensitivity 5-50 ng/mL). In the remaining 26% of the sample (n = 9), serum concentrations of psychotropic medications and/or active metabolites were detected. In each of these cases, infants had been exposed to the medication during pregnancy. Medications were not detected in infant serum when mothers had taken these agents solely during the postpartum period. No readily apparent difficulties with the infants were reported by mothers. CONCLUSIONS: These data support the low incidence of infant toxicity and adverse effects associated with antidepressant and benzodiazepine use during breastfeeding. These data also suggest that infant serum monitoring is helpful in the assessment of medication exposure in children of mothers who breastfeed while using psychotropic medications. Given the limited accumulated data regarding serum concentrations of psychotropic medications in breastfeeding infants, no single agent seems to be safer than another. Therefore, choice of pharmacologic treatment should be guided by the likelihood that it will result in restoration of maternal psychiatric well being. PMID- 10390298 TI - Previous exposure to measles, mumps, and rubella--but not vaccination during adolescence--correlates to the prevalence of pancreatic and thyroid autoantibodies. AB - OBJECTIVE: This study was designed to determine whether a relationship exists between previous exposure to measles, mumps, and rubella (MMR) by natural infection or vaccination or by new immunization with MMR vaccine, and either the presence or levels of autoantibodies against thyroid cell and pancreatic beta cell antigens. METHODS: Antibodies against MMR and autoantibodies against thyroglobulin, thyroid peroxidase, pancreas islet cells (ICA), islet cell surface, glutamic acid decarboxylase 65k autoantibodies, and insulin were studied before, and 3 months after, vaccination with combined MMR vaccine in 386 school children between 11 and 13 years of age. RESULTS: The vaccination changed neither the prevalence nor the level of autoantibodies. Children with rubella antibodies before vaccination had higher levels of ICA than did the rubella seronegative children. In contrast, thyroid autoantibody levels and prevalence were lower in children with antibodies against measles, mumps, or both before vaccination than in children without those antibodies. CONCLUSIONS: Previous natural infection or vaccination against measles, mumps, or both seemed to have an inhibitory effect on the development of thyroid autoantibodies. In contrast, children with previous exposure to rubella had higher levels of ICA. No evidence was found that MMR vaccination during adolescence may trigger autoimmunity. PMID- 10390299 TI - Procedural pain in newborn infants: the influence of intensity and development. AB - OBJECTIVE: Previous reports have shown that pain is managed inadequately in newborn infants. Ironically, clinicians believe that infants can experience pain much like adults, that infants are exposed daily to painful procedures, and that pain protection should be provided. In adults, a close relationship has been shown in how adults behave in response to pain, how painful they sense the stimulus to be, and physical measurements of the intensity of the stimulus. Whether similar parallels exist in newborn infants has not been examined. If these parallels do not exist in infants, it may help explain why clinicians fail to manage procedural pain in infants more effectively. The objective of this study was to determine whether the magnitude of infants' responses to nursing/medical procedures: 1) differs as a function of the invasiveness or intensity of the procedure; 2) differs as a function of intrauterine (gestational age at birth) and/or extrauterine (conceptional age) development; and 3) parallels the subjective pain ratings of clinicians for those procedures. METHODS: A broad developmental and clinical range of newborn infants was studied shortly before (baseline and preparatory periods), throughout, and shortly after (recovery period) required nursing/medical procedures during hospitalization. Heart rate, oxygen saturation, mean arterial pressure, and behavioral state (percentage of time spent in sleep or in agitation) were measured, and the magnitude of change in each in response to procedures was calculated. Procedures were categorized as mildly, moderately, and highly invasive to examine differences in response magnitude as a function of procedural invasiveness. Responses were compared as a function of prematurity and postnatal age. Clinicians' procedural pain ratings were compared with the magnitude of infants' responses. RESULTS: Of the original 152 infants, 135 were studied at least two times (range 2-27). Significant changes occurred in physiologic and behavioral measures in response to procedures indicative of pain responses. The magnitude of response generally increased with increased procedural invasiveness although there was considerable overlap of magnitude with invasiveness. Both premature and full-term infants differentiated procedural invasiveness. Very premature infants (<28 weeks' gestational age) exhibited increased increments in response magnitude with increasing postnatal age. Clinician's ratings of procedural painfulness were correlated with and predicted the magnitude of heart rate response to individual procedures. CONCLUSIONS: Similar to what has been shown in adults, newborn and developing infants show increased magnitude physiologic and behavioral responses to increasingly invasive procedures, demonstrating that even very prematurely born infants respond to pain and differentiate stimulus intensity. However, the considerable overlap of magnitude with invasiveness suggests that there is not a physiologic or behavioral threshold that clearly marks the presence of pain. Inconsistencies in physiologic and behavioral responses make reliance on a pain index difficult. The best approach may be one of universal precaution to provide pain management systematically to reduce the acute and long-term impact of early procedural pain. development, stimulus intensity, pain response. PMID- 10390300 TI - Effects of stroke localization on cardiac autonomic balance and sudden death. AB - BACKGROUND AND PURPOSE: Stroke has been shown to alter autonomic function. The purpose of this study was to show the differential effects of stroke localization on autonomic function parameters assessed by heart rate variability (HRV). METHODS: To determine the differential effect of ischemic stroke localization on autonomic cardiac innervation, we evaluated 62 patients with ischemic stroke and 62 age- and sex-matched controls. The localization of the infarct was determined by CT and MRI. Power spectrum analysis of HRV was performed. Myocardial necrosis was ruled out by echocardiography and creatine kinase myocardial isoenzymes measurements. RESULTS: All stroke patients had significantly decreased low frequency, high frequency, and standard deviation of all relative risk intervals values (P<0.001). However, patients with right-middle cerebral artery (R-MCA) and insula lesions had significantly lower power spectrum analysis values compared with all other localizations (P<0.001). In addition, 5 patients with R-MCA insular lesions died suddenly compared with 2 patients with left-middle cerebral artery insular lesions during hospitalization. Both sympathetic- and parasympathetic-controlled HRV were decreased in patients with ischemic stroke. The most pronounced decrease was found in the territory of R-MCA insular cortex, which suggests that cardiac autonomic tone may be regulated by insula and that these patients are more prone to cardiac complications such as arrhythmias and sudden death due to autonomic imbalance. CONCLUSION: We conclude that stroke in the region of insula (especially the right) leads to decreased HRV and to increased incidence of sudden death. PMID- 10390301 TI - Excess stroke among hypertensive men and women attributable to undertreatment of hypertension. AB - BACKGROUND AND PURPOSE: Most population-based studies indicate that a considerable proportion of hypertensive subjects are undertreated and that undertreatment is more prevalent among hypertensive men than among hypertensive women. The aim of our study was to investigate the consequences of undertreatment of hypertension for women and men in terms of stroke occurrence. METHODS: Approximately 45 000 men and women aged >/=20 years were examined in 2 population based studies in the Netherlands. A cohort of 2616 hypertensive subjects (pharmacologically treated hypertensives and untreated hypertensives who needed pharmacological treatment according to the severity of their hypertension and the coexistence of additional cardiovascular risk factors) was selected for a follow up study. Follow-up (mean duration, 4.6 years) was complete for 2369 (91%) of the enrolled hypertensive subjects. RESULTS: Compared with treated and controlled hypertensives, the relative risks of stroke for treated and uncontrolled hypertensives and for untreated hypertensives who needed treatment were 1.30 (95% CI, 0.70 to 2.44) and 1.76 (95% CI, 1.05 to 2.94), respectively. These relative risks and the prevalence of (undertreated) hypertension in the total population of 45 000 subjects were used to estimate the number of strokes in the Netherlands attributable to undertreatment. Among hypertensive men and women aged >/=20 years in the Netherlands, the proportions of strokes attributable to treated but uncontrolled blood pressure were 3.1% (95% CI, -5.2% to 18.7%) and 4.1% (95% CI, 7.2% to 20.7%), respectively. For untreated hypertensive men and women who should have been treated, these proportions were 22.8% (95% CI, 0.8% to 38.4%) and 25.4% (95% CI, 0. 5% to 42.5%), respectively. CONCLUSIONS: Increasing the detection of hypertension and improving adherence to current guidelines might prevent a considerable proportion of the incident strokes among hypertensives. The potential impact of achieving control of blood pressure in patients already being treated on the reduction of strokes requires further investigation. PMID- 10390302 TI - Recently occluded intracranial and extracranial carotid arteries. Relevance of the unstable atherosclerotic plaque. AB - BACKGROUND AND PURPOSE: It is now widely accepted that thrombotic coronary artery occlusion usually follows rupture of an unstable atherosclerotic plaque. The significance of such instability in arteries supplying the brain is less well appreciated. We therefore describe the clinical and pathological features of recent, symptomatic internal carotid artery occlusion to examine the pathogenetic role of plaque instability at both extracranial and intracranial sites. METHODS: Cases were selected from a consecutive series of 188 adult neuropathology autopsies. In 90 of these, the principal neuropathological abnormality was cerebral infarction, in 14 cases due to recent occlusion of 1 or more segments of the internal carotid artery. In each case, a full systemic, cardiovascular, and neuropathological autopsy was performed. Plaque instability was assessed by the presence or absence of a large, necrotic, lipid core; a thin, fibrous cap; and superficial inflammation. RESULTS: Of the 14 cases, 3 showed extracranial (carotid sinus), 7 intracranial, and 4 both extracranial and intracranial carotid artery occlusion. In 6 of the 7 occluded carotid sinuses, thrombus overlay an ulcerated, unstable, atherosclerotic plaque. In 1 extracranial and all 11 intracranial occlusions, there was either no atheroma or a mildly stenotic, stable, fibrous plaque, and in these cases, the cause of occlusion was embolism (8 cases), giant-cell arteritis (1 case), and unknown (3 cases). CONCLUSIONS: Coronary-type rupture of an unstable atherosclerotic plaque is the usual cause of fatal occlusion of the carotid sinus, but other causes usually underlie intracranial carotid occlusion. The nature and consequences of intracranial atherosclerosis require further study. PMID- 10390303 TI - Hemorrhagic transformation in acute ischemic stroke. The MAST-E study. MAST-E Group. AB - BACKGROUND AND PURPOSE: Hemorrhagic transformation (HT) is the most critical complication of thrombolytics in clinical trials in acute stroke. The aim of this study was to determine the rates and the predictors of HT in the Multicenter Acute Stroke Trial-Europe (MAST-E) study. METHODS: We performed a post hoc analysis of MAST-E data designed to assess the safety and efficacy of streptokinase administered intravenously within 6 hours of stroke onset. HT included all intracerebral hemorrhages and symptomatic hemorrhages (SHT) associated with clinical worsening. The predictors of HT and SHT were determined using multivariate modeling. RESULTS: Among the 310 patients included, 159 patients had HT and 37 SHT (97 and 33 in the streptokinase group and 62 and 4 in the placebo group, respectively). Patients with SHT had significantly more atrial fibrillation, diabetes mellitus, no heparin use, streptokinase treatment, and early CT signs. In the multivariate analysis, HT was predicted by early CT signs and streptokinase treatment. SHT was predicted by diabetes mellitus, early CT signs, streptokinase treatment, and the interaction between streptokinase treatment and decreased level of consciousness. Among the streptokinase-treated patients, the same predictors remained. CONCLUSIONS: The relative risks of HT after streptokinase were in the same range in MAST-E as in other streptokinase and tPA trials. Early CT signs were strong predictors of both HT and SHT, stressing that these patients are at high risk of bleeding. In our study, the predictors of HT and SHT were similar to those of tPA trials in acute stroke. PMID- 10390304 TI - Physical activity and ischemic stroke risk. The atherosclerosis risk in communities study. AB - BACKGROUND AND PURPOSE: The relationship between physical activity and stroke is inconclusive according to the 1996 US Surgeon General's Report on Physical Activity and Health. Therefore, this study examined the relationship between physical activity and ischemic stroke risk among 14 575 Atherosclerosis Risk in Communities Study participants aged 45 to 64 years free of self-reported stroke and coronary heart disease at baseline. METHODS: Eligible potential stroke hospitalizations were identified from ongoing hospital surveillance and from hospitalizations reported by the cohort study participants. All strokes were validated by hospitalization records. Physical activity was measured as sport, leisure (nonsport), and work with the use of the Baecke questionnaire. Multivariable Poisson and Cox proportional hazards models were used to determine the association of differing levels of physical activity with ischemic stroke incidence. RESULTS: During an average of 7.2 years of follow-up, 189 incident ischemic strokes occurred. Ischemic stroke incidence rates were highest in the lowest quartile of sport, leisure, and work scores. The hazard rate ratios with 95% CIs for ischemic stroke for the highest quartile compared with the lowest quartile of activity adjusted for age, sex, race-center, education, and smoking, were sport 0.83 (0.52, 1.32), leisure 0.89 (0.57, 1.37), and work 0.69 (0.47, 1.00). Further adjustment for factors that likely were intermediate variables (hypertension, diabetes, fibrinogen, and body mass index) between physical activity and stroke attenuated the associations. CONCLUSIONS: Our findings suggest that physical activity was weakly associated with a reduced risk of ischemic stroke among middle-aged adults. The association may be due to links between physical activity and other risk factors or due to chance. PMID- 10390305 TI - A systematic review of cost-effectiveness research of stroke evaluation and treatment. AB - BACKGROUND AND PURPOSE: This work was undertaken to review research addressing the cost-effectiveness of stroke-related diagnostic, preventive, or therapeutic interventions. METHODS: We performed searches of MEDLINE, Excerpta Medica online, HealthSTAR, and Sciences Citation Index Expanded and examined the reference lists of the studies and reviews obtained. From these, we selected studies that reported an incremental analysis of cost per effect, in which the effect measure was life-years or quality-adjusted life-years. We abstracted data from each study using a standardized reporting form. Twenty-six articles met the eligibility criteria and were included in the review. RESULTS: The methodological quality of the articles reviewed has improved compared with previously reported. Many stroke evaluation and treatment policies may result in benefits to health that are considered worth their cost. Some interventions were considered cost-ineffective (anticoagulation in low-risk nonvalvular atrial fibrillation and surveillance with duplex ultrasound after endarterectomy). Different studies addressing the cost-effectiveness of screening asymptomatic carotid stenosis resulted in strikingly divergent conclusions, from being cost-effective to being detrimental. Other studies omitted important costs that, if included, would likely have had profound impact on their cost-effectiveness estimates. CONCLUSIONS: Given the divergent conclusions drawn from studies addressing similar questions, it may be premature to use the results of cost-effectiveness research in developing stroke policy and practice guidelines. Successful implementation of such evaluations in the care of patients with stroke will depend on further standardization of methodology and critical appraisal of reported findings. PMID- 10390306 TI - Race, presenting signs and symptoms, use of carotid artery imaging, and appropriateness of carotid endarterectomy. AB - BACKGROUND AND PURPOSE: We sought to determine whether there are racial differences in use of carotid artery imaging after controlling for clinical factors and to ascertain racial differences in presenting signs and symptoms and overall appropriateness for carotid endarterectomy (CE). METHODS: We performed a retrospective cohort study of 803 patients older than 45 years, hospitalized between 1991 and 1994 at any of 4 Veterans Affairs Medical Centers, with a discharge diagnosis of transient ischemic attack or ischemic stroke. Clinical data were abstracted from the medical record, including presenting symptoms, diagnostic test results, and use of surgical procedures. Appropriateness for CE was determined according to RAND criteria. RESULTS: Black patients were more likely than white patients to present with stroke (78% versus 55%) but less likely to present with transient ischemic attack (22% versus 45%; P=0.001). There was no racial difference in medical comorbidity or preoperative risk. Black patients were less likely to have an imaging study of their carotid arteries (67% versus 79%; P=0.001). Race remained an independent predictor of imaging after adjustment for clinical factors (odds ratio=1.50; 95% CI, 1.06 to 2.13). Because of higher prevalence of significant carotid artery stenosis, whites were significantly more likely than blacks to be assessed as appropriate candidates for surgery with the use of RAND criteria (18% versus 4%; P=0.001). CONCLUSIONS: Use of carotid artery imaging, a critical step in determining eligibility for CE, is influenced by the patient's race after controlling for clinical presentation. Adjustment for appropriateness of CE reduces but does not eliminate the importance of race. PMID- 10390307 TI - Can stroke patients use visual analogue scales? AB - BACKGROUND AND PURPOSE: Visual analogue scales (VAS) have been used for the subjective measurement of mood, pain, and health status after stroke. In this study we investigated how stroke-related impairments could alter the ability of subjects to answer accurately. METHODS: Consent was obtained from 96 subjects with a clinical stroke (mean age, 72.5 years; 50 men) and 48 control subjects without cerebrovascular disease (mean age, 71.5 years; 29 men). Patients with reduced conscious level or severe dysphasia were excluded. Subjects were asked to rate the tightness that they could feel on the (unaffected) upper arm after 3 low pressure inflations with a standard sphygmomanometer cuff, which followed a predetermined sequence (20 mm Hg, 40 mm Hg, 0 mm Hg). Immediately after each change, they rated the perceived tightness on 5 scales presented in a random order: 4-point rating scale (none, mild, moderate, severe), 0 to 10 numerical rating scale, mechanical VAS, horizontal VAS, and vertical VAS. Standard tests recorded deficits in language, cognition, and visuospatial awareness. RESULTS: Inability to complete scales with the correct pattern was associated with any stroke (P<0.001). There was a significant association between success using scales and milder clinical stroke subtype (P<0.01). Within the stroke group, logistic regression analysis identified significant associations (P<0.05) between impairments (cognitive and visuospatial) and inability to complete individual scales correctly. CONCLUSIONS: Many patients after a stroke are unable to successfully complete self-report measurement scales, including VAS. PMID- 10390308 TI - Development of a stroke-specific quality of life scale. AB - BACKGROUND AND PURPOSE: Clinical stroke trials are increasingly measuring patient centered outcomes such as functional status and health-related quality of life (HRQOL). No stroke-specific HRQOL measure is currently available. This study presents the initial development of a valid, reliable, and responsive stroke specific quality of life (SS-QOL) measure, for use in stroke trials. METHODS: Domains and items for the SS-QOL were developed from patient interviews. The SS QOL, Short Form 36, Beck Depression Inventory, National Institutes of Health Stroke Scale, and Barthel Index were administered to patients 1 and 3 months after ischemic stroke. Items were eliminated with the use of standard psychometric criteria. Construct validity was assessed by comparing domain scores with similar domains of established measures. Domain responsiveness was assessed with standardized effect sizes. RESULTS: All 12 domains of the SS-QOL were unidimensional. In the final 49-item scale, all domains demonstrated excellent internal reliability (Cronbach's alpha values for each domain >/=0.73). Most domains were moderately correlated with similar domains of established outcome measures (r2 range, 0.3 to 0.5). Most domains were responsive to change (standardized effect sizes >0.4). One- and 3-month SS-QOL scores were associated with patients' self-report of HRQOL compared with before their stroke (P<0.001). CONCLUSIONS: The SS-QOL measures HRQOL, its primary underlying construct, in stroke patients. Preliminary results regarding the reliability, validity, and responsiveness of the SS-QOL are encouraging. Further studies in diverse stroke populations are needed. PMID- 10390309 TI - Improving outcomes for persons with aphasia in advanced community-based treatment programs. AB - BACKGROUND AND PURPOSE: Studies have yet to document that community-based aphasia treatment programs routinely produce results comparable or superior to published research protocols. We explore this issue here in an outcome study of individuals with aphasia enrolled in 2 community-based, comparably managed and equipped therapy programs, which use a specially designed computer-based tool that is employed therapeutically in adherence to an extensive, detailed, and formally trained patient care algorithm. METHODS: Patients (n=60) were assessed before and after treatment with standardized instruments at both the impairment and the disability levels. Pretreatment and posttreatment means were calculated and compared, with statistical significance of differences established with the use of 1-tailed matched t tests. One-way ANOVAs were used to analyze the comparability of patient performance changes among various subgroups, eg, patients in acute versus chronic stages of aphasia, patients by aphasia diagnostic type at start of care, patients by severity level at start of care, and patients by treatment location. RESULTS: Analysis shows that patients spanned a wide range of aphasia diagnostic types, impairment severity levels at start of care, and times after onset. Patients' mean performance scores improved significantly in response to treatment in all measures assessed at both the impairment level and the functional communication level. Mean overall improvements ranged from 6.6% to 19.8%, with statistical significance ranging from P=0.0006 to P<0.0001. ANOVAs revealed no significant differences between improvements in patients in the acute versus chronic stages of aphasia, between those at different impairment severity levels at start of care, between those treated at different locations, or, at the functional level, between those with different diagnostic types of aphasia at start of care. CONCLUSIONS: Measures of both language impairment and functional communication can be broadly, positively, and significantly influenced by therapy services that are delivered to persons with aphasia in these community-based programs. The significant improvements are shown to be available to individuals with chronic as well as acute aphasia and independent of diagnostic type of aphasia, impairment severity at start of care, or geographic program location. PMID- 10390310 TI - Middle cerebral artery stroke that includes the premotor cortex reduces mobility outcome. AB - BACKGROUND AND PURPOSE: The premotor cortex (PMC) (Brodmann 6) contributes uniquely to proximal upper and lower limb power and plays a role in the organization of motor behaviors. We assessed the degree to which PMC damage affected functional outcome. METHODS: We prospectively compared the functional outcome of patients with a first stroke in the middle cerebral artery distribution that either left the PMC intact (PMC-; n=19) or damaged the PMC (PMC+; n=12). The Functional Independence Measure for disability and the motor score of the Stroke Impairment Assessment Set for impairment assessed outcome. RESULTS: Demographic and clinical features and lesion volume were comparable for the PMC+ and PMC- groups. However, the PMC- group demonstrated significant gain in mobility and in proximal leg movement. This focal improvement contributed to the trend in the PMC- group toward greater independent ambulation. CONCLUSIONS: Decreased motor recovery of proximal lower limbs in humans with PMC damage supports the idea that it is the origin of corticoreticulospinal pathways that subserve proximal lower extremity function. Furthermore, persistent proximal weakness after PMC damage may amplify other motor impairments, which include defects in planning, initiating, and sequencing. Neurorehabilitation outcomes may contribute to a more detailed functional anatomy after stroke and partial recovery. PMID- 10390311 TI - Electrical stimulation of wrist extensors in poststroke hemiplegia. AB - BACKGROUND AND PURPOSE: It has been suggested that cyclic neuromuscular electrical stimulation (ES) may enhance motor recovery after stroke. We have investigated the effects of ES of the wrist extensors on impairment of wrist function and on upper-limb disability in patients being rehabilitated after acute stroke. METHODS: We recruited 60 hemiparetic patients (mean age, 68 years) 2 to 4 weeks after stroke into a randomized, controlled, parallel-group study comparing standard rehabilitation treatment with standard treatment plus ES of wrist extensors (3 times 30 minutes daily for 8 weeks). Isometric strength of wrist extensors was measured using a device built for that purpose. Upper-limb disability was assessed with use of the Action Research Arm Test (ARAT). Observations were continued for 32 weeks (24 weeks after the finish of ES or the control intervention phase). RESULTS: The change in isometric strength of wrist extensors (at an angle of 0 degrees extension) was significantly greater in the ES group than the control group at both 8 and 32 weeks (P=0.004, P=0.014 by Mann Whitney U test). At week 8 the grasp and grip subscores of the ARAT increased significantly in the ES group compared with that in the control group (P=0.013 and P=0.02, respectively); a similar trend was seen for the total ARAT score (P=0.11). In the subgroup of 33 patients with some residual wrist extensor strength at study entry (moment at 0 degrees extension >0), the ARAT total score had increased at week 8 by a mean of 21.1 (SD, 12.7) in the ES group compared with 10.3 (SD, 9.0) in the control group (P=0.024, Mann Whitney U test); however, at 32 weeks the differences between these 2 subgroups were no longer statistically significant. CONCLUSIONS: ES of the wrist extensors enhances the recovery of isometric wrist extensor strength in hemiparetic stroke patients. Upper-limb disability was reduced after 8 weeks of ES therapy, with benefits most apparent in those with some residual motor function at the wrist. However, it is not clear how long the improvements in upper-limb disability are maintained after ES is discontinued. PMID- 10390312 TI - Prevalence of intracranial saccular aneurysms in a Japanese community based on a consecutive autopsy series during a 30-year observation period. The Hisayama study. AB - BACKGROUND AND PURPOSE: Subarachnoid hemorrhage is a life-threatening disease that occurs mostly because of the rupture of intracranial saccular aneurysms. However, little is known about the prevalence of ruptured and unruptured aneurysms in the general population. The aim of the present study was to examine the prevalence of intracranial aneurysms on the basis of a consecutive autopsy series over a 30-year observation period in a general Japanese population in Hisayama. METHODS: We evaluated 1230 consecutive autopsy cases with craniotomy among the total deaths of Hisayama residents during 1962 through 1991 (overall autopsy rate, 80.1%). RESULTS: A total of 73 intracranial saccular aneurysms were found in 57 cases (4.6%). The prevalence of aneurysms for women was 2.4 times higher than that for men (7.1% versus 2.9%). Among men, the prevalence of aneurysms remained unchanged across the range of age groups. In contrast, there were 2 peaks in the prevalence of aneurysms for women falling in the 40- to 49 year (14.3%) and 60- to 69-year age groups (14.5%). The most common site of the aneurysms was the middle cerebral artery (31.5%), followed by the anterior communicating artery (30.1%), anterior cerebral artery (15.1%), vertebrobasilar artery (12.3%), and internal carotid artery (11.0%). Among these 73 aneurysms, 29 (39.7%) were ruptured. Ruptured aneurysms were common in subjects <80 years of age, whereas unruptured aneurysms were prevalent in those >/=80 years of age. The frequency of ruptured aneurysms was highest in the vertebrobasilar system (66.7%) and lowest in the middle cerebral artery (13.0%). CONCLUSIONS: Our data suggest that intracranial aneurysms are more frequent in women in the general Japanese population. Aneurysms are more prevalent in the middle cerebral artery, but the risk of rupture is highest in the vertebrobasilar system. PMID- 10390313 TI - Structural fragility and inflammatory response of ruptured cerebral aneurysms. A comparative study between ruptured and unruptured cerebral aneurysms. AB - BACKGROUND AND PURPOSE: Despite technical advances in endovascular and microsurgical treatment, patients with aneurysmal subarachnoid hemorrhage still have a high mortality and morbidity rate. To improve the treatment results in patients with aneurysms, we must better understand the pathophysiology of cerebral aneurysms and the mechanisms leading to their rupture. Therefore, we studied the pathological differences between unruptured and ruptured aneurysms. METHODS: Ruptured (n=44) and unruptured (n=27) aneurysms were obtained at surgery. The aneurysmal endothelium was scored from 0 (normal) to 5 (complete disruption) by using a scanning electron microscope. The aneurysmal wall was evaluated by immunohistochemical methods. The wall structure was scored from 1 (dense collagen and rich, smooth muscle cells) to 5 (hyaline-like structure). The degree of inflammatory cell invasion into the wall was also scored from 0 (very few cells) to 3 (many cells). RESULTS: Ruptured aneurysms manifested significant endothelial damage (score of 3.7 versus 0.8; Mann-Whitney U test, P<10(-3)), significant structural changes of the wall (3.7 versus 1.7, P<10(-5)), and significant inflammatory cell invasion (2.2 versus 0.8, P<10(-4)) compared with unruptured aneurysms. There was a significant correlation between the score for wall structure and the score for inflammatory cell invasion (Rs=0. 63; Spearman rank correlation test, P<10(-5)). The pathophysiology of several symptomatic unruptured aneurysms was similar to that of ruptured aneurysms. CONCLUSIONS: We conclude that the pathophysiology of unruptured, asymptomatic and ruptured aneurysms is different. The wall of ruptured aneurysms was found to be fragile, possibly because macrophage infiltration into the aneurysmal wall resulted in loss of smooth muscle cells and in degradation of matrix proteins. PMID- 10390314 TI - Multivariate analysis of predictors of cerebral vasospasm occurrence after aneurysmal subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: The role of type of treatment on cerebral vasospasm occurrence after aneurysmal subarachnoid hemorrhage (SAH) has not been studied. Through multivariate analysis we determined the independent prognostic factors of the occurrence of symptomatic vasospasm following aneurysmal SAH in a study cohort of 244 patients undergoing either surgical or endovascular treatment. The prognostic factors of sequelae after aneurysmal SAH were studied as well. METHODS: Symptomatic vasospasm was defined as the association of deterioration in a patient's neurological condition between 3 and 14 days after SAH with no other explanation and an increase in mean transcranial Doppler velocities of >120 cm/s. The prognostic factors were registered on admission and during the intensive care stay. RESULTS: Symptomatic vasospasm occurred in 22.2% surgical patients compared with 17.2% endovascular treatment patients (P=0.37). Multivariate analysis revealed that the probability of occurrence of symptomatic vasospasm decreased with age >50 years (relative risk [RR], 0.47 [0.25 to 0.88]) and severe World Federation of Neurological Surgeons (WFNS) grade measured on admission (RR, 0.43 [0.20 to 0.90]) and increased with hyperglycemia occurring during the intensive care stay (RR, 1.94 [1.04 to 3.63]). No difference in risk of symptomatic vasospasm could be identified between surgical and endovascular treatment. Symptomatic vasospasm (OR, 4.73 [CI, 1. 77 to 12.6]) as well as WFNS grade of >2 (OR, 8.95 [3.46 to 23.2]), treatment complications (OR, 8.39 [3.16 to 22.3]), and secondary brain insults were associated with an increased risk of 6-month sequelae. CONCLUSIONS: Age <50 years, good neurological grade, and hyperglycemia were all associated with an increased risk of cerebral vasospasm whereas treatment was not. This provides a basis for future clinical prospective randomized trials comparing both treatments. PMID- 10390315 TI - Safety of intrathecal sodium nitroprusside for the treatment and prevention of refractory cerebral vasospasm and ischemia in humans. AB - BACKGROUND AND PURPOSE: The delayed type of cerebral vasoconstriction known as cerebral vasospasm (DCV) remains an important cause of permanent neurological injury and death following aneurysmal subarachnoid hemorrhage despite best current medical therapy. The mechanism of DCV remains unknown. A new treatment for refractory DCV using intrathecally delivered sodium nitroprusside and results in 21 patients is reported. METHODS: Candidates for treatment were patients with secured cerebral aneurysms presenting with clinical or radiographic SAH of grade 3 or higher. Patients with and without established DCV were treated. In 57% (12/21 patients) the diagnosis of severe DCV refractory to conventional treatment (HHH therapy and nimodipine) was established before treatment. Ten patients received ITSNP prophylactically. All patients with established DCV were in grave neurological condition before treatment. Procedures for vasospasm reversal were performed under simultaneous angiographic control with extensive hemodynamic and neurophysiologic monitoring. ITSNP was delivered by intraventricular or subdural catheter or by direct intraoperative suffusion. End points of intervention for established DCV were (1) durable angiographic reversal of vasoconstriction, (2) failure to effect reversal within 30 minutes, and (3) adverse effect. End points for DCV prevention were (1) post-SAH day 10 without evidence of vasoconstriction and (2) adverse effect. Cerebral angioplasty was used concomitantly in 9 treatments. The total number of treatments recorded was 171. RESULTS: The overall neurological outcome was good or excellent in 76% of patients (16/21) overall and in 88.9% of patients (16/18) having at least a 1-month follow-up. Of the 5 patients with less-than-good outcome, 4 had presented initially with severe neurological injury (clinical SAH grade 4). Angiography demonstrated reversal or amelioration of vasoconstriction in 83% (5/6 cases) of established DCV treated by ITSNP alone. Among patients treated prophylactically, none developed clinical DCV. CONCLUSIONS: These results suggest that ITSNP is a safe and potentially effective treatment for established DCV and cerebral ischemia refractory to conventional treatment. The preliminary results of prophylactic treatment are also favorable with regard to safety. PMID- 10390317 TI - Uncoupling of oxygen and glucose metabolism in persistent crossed cerebellar diaschisis. AB - BACKGROUND AND PURPOSE: The pathophysiology of deafferentation-induced changes after stroke remains unclear. Some supratentorial strokes cause persistent decreases in blood flow and metabolism in the contralateral cerebellum (persistent crossed cerebellar diaschisis[CCD]). Our previous study showed uncoupling of oxygen consumption and blood flow in this condition, which may reflect a characteristic change in brain metabolism caused by deafferentation. This uncoupling might be related to oxidation of some substrates other than blood borne glucose, which could also lead to the uncoupling of oxygen consumption and glucose utilization. The purpose of this study was to investigate whether oxygen consumption is uncoupled from glucose utilization in persistent CCD. METHODS: Using positron emission tomography in 10 unilateral supratentorial stroke patients, we evaluated regional blood flow, oxygen consumption, and glucose utilization in the cerebellar cortex in the chronic stage. Eight patients with a significant cerebellar blood flow asymmetry, defined as outside the 95% confidence limits predefined in 9 normal subjects, were selected as patients with persistent CCD. RESULTS: In patients with CCD, the cerebellar cortex contralateral to the stroke showed significant decreases in both oxygen consumption and glucose utilization compared with the ipsilateral cerebellar cortex. The decrease in oxygen consumption was less than the decrease in glucose utilization, resulting in a significant increase in the oxygen consumption/glucose utilization ratio. CONCLUSIONS: Persistent CCD caused by stroke may induce uncoupling of oxygen consumption and glucose utilization, which may reflect a characteristic change in brain metabolism caused by deafferentation. PMID- 10390316 TI - Nimodipine and perfusion changes after stroke. AB - BACKGROUND AND PURPOSE: Meta-analysis of previous trials of oral nimodipine in acute stroke has suggested a benefit when commenced within 12 hours of onset. We sought to study the effect of oral nimodipine on reperfusion after acute stroke and the relation between reperfusion and outcome. METHODS: Fifty patients with acute middle cerebral artery territory cortical infarction were blindly randomized within 12 hours of onset to either oral nimodipine (30 mg every 6 hours) or placebo. Treatment was continued for 2 weeks. Cerebral blood flow was assessed with the use of 99mTc-hexamethylpropyleneamine oxime single-photon emission CT before therapy, 24 hours later, and at 3 months. Hypoperfusion was measured by a validated volumetric technique. Neurological impairment and functional outcome were assessed with the Canadian Neurological Scale and Barthel Index, respectively. Tissue loss was measured with CT at 3 months. Four patients were excluded from analysis for technical reasons. RESULTS: Twenty-three patients received nimodipine, and 23 received placebo. In the nimodipine group, there was early reperfusion that was not maintained at outcome (P=0.01). In the placebo group, mean infarct hypoperfusion volumes showed no overall change. Nonnutritional reperfusion in nimodipine-treated patients was associated with adverse neurological (P=0.05) and functional outcome (P=0.06). There was, however, no difference in clinical outcome between the 2 groups. CONCLUSIONS: Oral nimodipine administered within 12 hours enhanced acute reperfusion, but this was largely nonnutritional. Larger studies using a shorter treatment delay are required to evaluate the clinical efficacy of nimodipine in acute ischemic stroke. PMID- 10390318 TI - A decrease in regional cerebral blood volume and hematocrit in crossed cerebellar diaschisis. AB - BACKGROUND AND PURPOSE: The pathophysiology of crossed cerebellar diaschisis (CCD) remains to be elucidated. In CCD, the metabolic suppression resulting from deafferentation may cause vasoconstriction, which may result in a decrease in cerebral blood volume (CBV) and may differentially affect the flows of red blood cells and of plasma. The purpose of this study was to investigate whether CCD decreases the total CBV (cerebral red blood cell volume [CRCV] plus cerebral plasma volume [CPV]) and, if so, whether CCD differentially affects the CRCV and CPV, resulting in a change in hematocrit. METHODS: We used positron emission tomography to study 7 patients with a unilateral supratentorial infarct and CCD. The distributions of CRCV and CPV were assessed by using 15O-labeled carbon monoxide and 62Cu-labeled human serum albumin-dithiosemicarbazone tracers, respectively. The CRCV, CPV, and calculated hematocrit values were compared between the cerebellar hemispheres. RESULTS: In the cerebellar cortex contralateral to the supratentorial infarct, the values of CRCV, CPV, and total CBV were significantly decreased compared with those in the ipsilateral cerebellar cortex. The CRCV was decreased to a greater degree than the CPV, and the value of the hematocrit was decreased in the contralateral cerebellar cortex. CONCLUSIONS: CCD may decrease the total CBV, which may reflect vasoconstriction caused by decreased metabolism due to deafferentation. In addition, the more pronounced decrease in CRCV than in CPV may result in a decrease in hematocrit in CCD. PMID- 10390319 TI - Cerebral hemodynamics in relation to patterns of collateral flow. AB - BACKGROUND AND PURPOSE: We sought to investigate the relation between collateral flow via different pathways and hemodynamic parameters measured by dynamic susceptibility contrast-enhanced MRI in patients with severe carotid artery disease. METHODS: Dynamic susceptibility contrast-enhanced MRI was performed in 66 patients and 33 control subjects. Patients had severe stenosis (>70%, n=12), unilateral occlusion (n=38), or bilateral occlusion (n=16) of the internal carotid artery (ICA). Cerebripetal flow and collateral flow via the circle of Willis were investigated with MR angiography. Collateral flow via the ophthalmic artery was investigated with transcranial Doppler sonography. RESULTS: Patients with ICA stenosis had well-preserved cerebral perfusion and were in general not dependent on collateral supply. Patients with unilateral ICA occlusion had impaired cerebral perfusion. However, appearance time, peak time, and mean transit time in white matter were less increased in patients with than in patients without collateral flow via the circle of Willis (P<0.05). Furthermore, patients with collateral flow via both anterior and posterior communicating arteries had less increased regional cerebral blood volume than patients with collateral flow via the posterior communicating artery only (P<0.05). Patients with bilateral ICA occlusion had severely compromised hemodynamic status despite recruitment of collateral supply. CONCLUSIONS: In patients with unilateral ICA occlusion, the pattern of collateral supply has significant influence on hemodynamic status. Collateral flow via the anterior communicating artery is a sign of well-preserved hemodynamic status, whereas no collateral flow via the circle of Willis or flow via only the posterior communicating artery is a sign of deteriorated cerebral perfusion. PMID- 10390320 TI - Asymptomatic embolization predicts stroke and TIA risk in patients with carotid artery stenosis. AB - BACKGROUND AND PURPOSE: Improved methods of identifying patients at high risk of thromboembolism would allow improved targeting of therapy. One such situation is carotid artery stenosis. This is associated with an increased risk of stroke, which can be reduced by carotid endarterectomy. However, the risk-benefit ratio is low in patients with tight asymptomatic stenosis and moderate symptomatic stenosis. Most stroke in patients with carotid stenosis is believed to be embolic. Therefore, the detection of asymptomatic cerebral emboli using Doppler ultrasound may allow identification of a high-risk group. METHODS: Transcranial Doppler ultrasound was used to record for 1 hour the ipsilateral middle cerebral artery in 111 patients with >60% carotid artery stenosis (69 symptomatic, 42 asymptomatic). The Doppler audio signal was recorded onto digital audio tape for later analysis for embolic signals (ES) by an individual blinded to clinical details. In 67 subjects the relationship between ES and angiographically determined plaque ulceration was investigated. All subjects were followed up prospectively, and the relationship between ES and risk of future ipsilateral carotid artery territory ischemic events (TIA and stroke) was determined. RESULTS: ES were detected in 41(36.9%) subjects. In symptomatic patients there was a significant inverse relationship between the number of ES per hour and time elapsed since last symptoms (Spearman's rho=-0.2558, P=0.034). ES were more common in subjects with plaque ulceration, with a relative risk of 4. 94 (95% CI, 1.23 to 19.84; P=0.025) after controlling for both symptomatic status and degree of stenosis. The presence of ES at entry was predictive of TIA and stroke risk during follow up in both symptomatic (P=0.02) and asymptomatic patients (P=0.007). Considering all 111 patients, the presence of asymptomatic embolization was predictive of a further ischemic event, with an adjusted OR of 8.10 (95% CI, 1.58 to 41.57; P=0.01) after controlling for other cardiovascular risk factors, degree of stenosis, symptomatic status, and aspirin or warfarin use. CONCLUSIONS: Asymptomatic embolization in patients with carotid artery stenosis correlates with known markers of increased stroke risk and is an independent predictor of future stroke risk in patients with both symptomatic and asymptomatic carotid stenosis. It may allow identification of a high-risk group of patients who will particularly benefit from carotid endarterectomy. A large multicenter study is now required to confirm these findings. PMID- 10390321 TI - Reliability and validity of noninvasive imaging of internal carotid artery pseudo occlusion. AB - BACKGROUND AND PURPOSE: Our study evaluated noninvasive tests for the diagnosis of atheromatous internal carotid artery (ICA) pseudo-occlusion. METHODS: Twenty patients (17 men, 3 women; mean age +/-SD, 64.3+/-11.6 years) with angiographically proven atheromatous ICA pseudo-occlusion (20 vessels) were prospectively examined with MR angiography (MRA; 2D and 3D time-of-flight techniques), color Doppler-assisted duplex imaging (CDDI) and power-flow imaging (PFI) with and without an intravenous ultrasonic contrast agent. As a control group, 13 patients (13 men; mean+/-SD age, 63.0+/-9.0 years) with angiographically proven ICA occlusion (13 vessels) were studied with the same techniques. For the determination of interobserver agreement (kappa statistics), the findings of each diagnostic technique were read by 2 blinded and independent observers who were not involved in patient recruitment and initial data acquisition. Specificity and sensitivity were calculated for all noninvasive techniques (observer consensus) in comparison to the standard of reference (intra arterial angiography). RESULTS: Interobserver reliabilities were kappa=0.86 for intra-arterial angiography, kappa=0.90 for unenhanced CDDI, kappa=0. 93 for enhanced CDDI, kappa=0.93 for unenhanced PFI, kappa=1.0 for enhanced PFI, kappa=0.93 for 2D MRA, and kappa=0.77 for 3D MRA, respectively (P<0.0001). Specificities and sensitivities were 0.92 and 0.70 for unenhanced CDDI, 0.92 and 0.83 for enhanced CDDI, 0.92 and 0.95 for unenhanced PFI, 1.0 and 0.94 for enhanced PFI, 1.0 and 0.65 for 2D MRA, and 0.89 and 0.47 for 3D MRA, respectively. CONCLUSIONS: Advanced ultrasonographic techniques, especially PFI (with only 1 false-positive diagnosis of occlusion in the present series), can provide reliable and valid data to differentiate between ICA pseudo-occlusion and complete occlusion. In contrast, time-of-flight MRA at its present state is not capable of predicting minimal residual flow within a nearly occluded ICA. PMID- 10390322 TI - Success rate of transcranial color-coded duplex ultrasonography in visualizing the basal cerebral arteries in vascular patients over 60 years of age. AB - BACKGROUND AND PURPOSE: Clinically important atherosclerotic cerebrovascular disease is mainly found in patients aged >60 years. Transcranial color-coded duplex ultrasonography (TCCD) is a relatively new technique for investigating the basal cerebral arteries; however, it is often hampered by impenetrable ultrasound windows. The aim of this study was to ascertain the as yet unknown success rate of TCCD regarding visualization of the basal cerebral arteries in patients >60 years, to provide reference data, and to compare any possible male/female differences. METHODS: In 112 atherosclerotic white patients >60 years of age, the anterior, middle, and posterior cerebral arteries and the vertebral and basilar arteries were insonated. RESULTS: In men, 99% of the temporal and 94% of the suboccipital windows could be penetrated by ultrasound compared with 77% and 95%, respectively, in women. The male versus female vessel detection rates were 91% versus 58% for the anterior cerebral artery, 97% versus 73% for the middle cerebral artery, 97% versus 68% for the posterior cerebral artery, 94% versus 93% for the vertebral artery, and 91% versus 79% for the basilar artery. In 77% of men but only 33% of women could all vascular segments be investigated. All intracranial arteries were insonated at a deeper level in men. The women showed significantly higher blood flow velocities than the men. CONCLUSIONS: In elderly white men the vessel detection rate is >90%. In women there is a much lower detection rate, due to impenetrable temporal windows. Visualization of all major intracranial arteries is possible in only one third of female patients >60 years of age. PMID- 10390323 TI - Decompressive craniectomy, reperfusion, or a combination for early treatment of acute "malignant" cerebral hemispheric stroke in rats? Potential mechanisms studied by MRI. AB - BACKGROUND AND PURPOSE: Both early reperfusion and decompressive craniectomy have proved beneficial in the treatment of large space-occupying "malignant" hemispheric stroke. The aim of this study was to directly compare the benefit of reperfusion with that of craniectomy and to study the effects of combined treatment in a rat model of focal cerebral ischemia. METHODS: Cerebral ischemia was introduced in 28 rats. Four groups were investigated: (1) no treatment, (2) decompressive craniectomy, (3) reperfusion, and (4) reperfusion and craniectomy as treatment at 1 hour after middle cerebral artery occlusion. Perfusion- and diffusion-weighted MRI were performed serially from 0.5 to 6 hours after middle cerebral artery occlusion. RESULTS: The 6-hour DWI-derived hemispheric lesion volumes in the reperfusion group (10.2+/-3.9%), the craniectomy group (23.0+/ 6.4%), and the combination group (21.8+/-12.4) were significantly smaller than that in the control group (44.1+/-5.4%) (P<0.05). Reperfusion, craniectomy, and combined treatment led to higher perfusion in the cortex compared with the control group, whereas only reperfused animals achieved significantly higher perfusion in the basal ganglia. In 5 animals, combined reperfusion and decompressive craniectomy resulted in an early contrast media enhancement. CONCLUSIONS: Early reperfusion and craniectomy were shown to be effective in decreasing infarction volume by improving cerebral perfusion. Reperfusion remains the best therapy in malignant hemispheric stroke. Combined treatment yields no additional benefit compared with single treatment, probably because of early blood-brain barrier breakdown. PMID- 10390324 TI - Effects of citicoline combined with thrombolytic therapy in a rat embolic stroke model. AB - BACKGROUND AND PURPOSE: We sought to evaluate the effects of the combination of cytidine-5'-diphosphocholine (citicoline) and thrombolysis on infarct size, clinical outcome, and mortality in a rat embolic stroke model. METHODS: Eighty three Sprague-Dawley rats were embolized in the carotid territory with a single fibrin embolus and randomly assigned to the following treatment groups: (1) control (saline), (2) citicoline 250 mg/kg, (3) citicoline 500 mg/kg, (4) recombinant tissue plasminogen activator (rtPA) 5 mg/kg, (5) rtPA 5 mg/kg plus citicoline 250 mg/kg, and (6) rtPA 5 mg/kg plus citicoline 500 mg/kg. rtPA was administered as a continuous intravenous infusion over 45 minutes starting 45 minutes after embolization; citicoline was given intraperitoneally 30 minutes and 24, 48, and 72 hours after embolization. At 96 hours, the brains were fixed and stained by hematoxylin-eosin, and infarct volumes were measured. Neurological scores were determined daily. RESULTS: The median infarct size, measured as percentage of the affected hemisphere, in the control group was 37% (interquartile range, 26% to 69%) compared with 22% (5% to 52%; P=NS) in group 2, 11% (5% to 34%; P=NS) in group 3, 24% (12% to 31%; P=NS) in group 4, 11% (3% to 22%; P=0.02) in the combined group 5, and 19% (9% to 51%; P=NS) in group 6. The infarct size was significantly reduced in the combined citicoline+rtPA-treated groups to a median of 13% (5% to 30%; P<0.01). Citicoline 500 mg/kg and citicoline combined with rtPA also promoted functional recovery. CONCLUSIONS: These results demonstrate that the combination of low-dose citicoline and rtPA significantly reduced infarct size in this focal ischemia model. PMID- 10390325 TI - The neuroprotective effect of the novel AMPA receptor antagonist PD152247 (PNQX) in temporary focal ischemia in the rat. AB - BACKGROUND AND PURPOSE: Evidence suggests that glutamate contributes to ischemic brain damage through activation of the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptor. We tested the novel, selective AMPA receptor antagonist PD152247 (PNQX) in a model of temporary focal ischemia to determine the dose-response relationship and to investigate the contribution of drug induced hypothermia to the neuroprotective action of AMPA receptor antagonists. METHODS: Temporary focal cerebral ischemia was induced in Sprague-Dawley rats by occluding the middle cerebral artery and both carotid arteries for 3 hours. Body temperature was monitored by telemetry. PNQX was administered intraperitoneally or by intravenous infusion with various doses for 6 hours. Lesion volume was determined after 3 days by stereological methods. RESULTS: PNQX reduced the lesion volume by 51% after intraperitoneal administration. The intravenous dose response study demonstrated that the lowest effective dose of PNQX was 1.0 mg/kg per hour, which corresponded to a steady state plasma level of 685 ng/mL. Neuroprotection was demonstrated at PNQX plasma concentrations that did not lower body temperature over the entire course of the experiment. CONCLUSIONS: AMPA receptor activation plays an important role in the development of ischemic brain damage. Thus, novel AMPA receptor antagonists may be useful for the treatment of stroke in humans. PMID- 10390326 TI - Family caregiving for patients with stroke. Review and analysis. AB - BACKGROUND: The literature on family caregiving for stroke patients is reviewed with the goals of (1) evaluating the effects of stroke caregiving on caregivers' well-being, (2) outlining deficiencies and methodological limitations of current research, and (3) outlining policy and practice implications of current studies. SUMMARY OF REVIEW: A total of 20 published stroke caregiving research articles were included in this review. Across studies, the effects of stroke caregiving on caregivers' well-being and the significant predictors of caregivers' depression were analyzed. Current evidence suggests that stroke caregivers have elevated levels of depression at both the acute stroke phase and the chronic stroke phase. However, major gaps are apparent in this literature, with few studies addressing such areas as caregiver physical health, ethnicity, and caregiver interventions. CONCLUSIONS: Given the increasing prevalence of stroke as well as the increasing pressures on families to provide care, more research is needed to guide policy and practice in this understudied topic. PMID- 10390327 TI - Which targets are relevant for therapy of acute ischemic stroke? AB - BACKGROUND: The efficiency of various strategies of neuroprotection is well documented in animal experiments but is thus far disappointing in ischemic stroke, for which only early reperfusion induced by thrombolysis has improved clinical outcome. This discrepancy between expectation from experimental research and clinical reality may be related to differences in the pathogenetic factors contributing to infarction. SUMMARY OF COMMENT: Positron emission tomography cerebral blood flow studies within 3 hours of onset were used to identify the various compartments of the infarct outlined on MRI 2 to 3 weeks after a hemispheric stroke in 10 patients. Critical hypoperfusion below the viability threshold accounted for the largest proportion (mean, 70%) of the final infarct, whereas penumbral tissue (18%) and initially sufficiently perfused tissue (12%) were responsible for considerably smaller portions of the final infarct. CONCLUSIONS: These results indicate that early critical flow disturbance leading to rapid cell damage is the predominant cause of infarction, while secondary and delayed pathobiochemical processes in borderline or initially sufficiently perfused regions contribute only little to the final infarct. Therefore, emerging therapeutic strategies should be targeted to the initially critically perfused tissue subcompartments. Clinical drug trials might benefit from stratification of patients for target tissue compartments applying functional imaging. PMID- 10390328 TI - Types of recurrent stroke in survivors of intracerebral hemorrhage. PMID- 10390329 TI - SPECT-derived relative perfusion defect and CT-derived hypodense region in acute intracerebral hemorrhage. PMID- 10390330 TI - Ataxic hemiparesis: an old debate for Lacune aficionados. PMID- 10390332 TI - Abstracts of literature PMID- 10390331 TI - Microembolic signals and early recurrent cerebral ischemia in carotid artery disease. PMID- 10390333 TI - The ultraflex diamond stent for malignant biliary obstruction. AB - The Ultraflex Diamond stent presents three features that might be of interest when treating biliary strictures: ease of insertion, high flexibility, and presentation of relatively large meshes. Limited clinical studies have shown its efficacy in relieving duct obstruction (in contrast with some other stent models, correct positioning of the Ultraflex Diamond stent provides adequate biliary drainage in almost all cases), and satisfactory long-term patency rates. The main improvement awaited is an efficient means to prevent late stent obstruction. The addition of coverage to this stent is feasible and could be the first step toward this goal. PMID- 10390334 TI - The ultraflex diamond stent for benign biliary obstruction. AB - The most frequent type of benign bile duct stricture (BBDS) that complicates surgery, is not an indication for placement of any current model of metal stent because of poor long-term results. This statement could not apply to less frequent types of BBDSs, in particular those complicating chronic pancreatitis (CP) and radiotherapy. The authors report their preliminary results observed in 10 patients whose CP-related BBDS was treated with an Ultraflex Diamond stent. No stent dysfunction was detected in the observed patients during a mean follow-up of 17 months. Studies involving larger numbers of patients followed for longer periods of time are necessary to know whether endoscopy using this material may replace surgery in this indication. PMID- 10390335 TI - Expandable metal stents for benign pancreatic duct obstruction. AB - Among patients with advanced chronic pancreatitis and morphologically demonstrable main pancreatic duct (MPD) abnormalities who are candidates for endotherapy, about two-thirds have a MPD stricture and require stenting to achieve appropriate ductal decompression. The standard stent used in this indication is the plastic stent, which provides a median patency rate of 6 to 12 months. The requirement for stent exchange represents a limitation for the treatment of this relatively young patient population. By analogy with the biliary tract, the authors hypothesize that self-expandable metal stents might offer a long-lasting drainage of the MPD. Several years ago the authors used such stents on an experimental basis to treat patients with MPD strictures. Although routine use of self-expandable metal stents should be discouraged, the authors propose some guidelines for further technical improvement. PMID- 10390336 TI - Structural compensation in an archaeal selenocysteine transfer RNA. AB - A new type of structural compensation between the lengths of two perpendicularly oriented RNA double helices was found in the archaeal selenocysteine tRNA from Methanococcus jannascii. This tRNA contains only four base-pairs in the T-stem, one base-pair less than in all other cytosolic tRNAs. Our analysis shows that such a T-stem in an otherwise normal tRNA cannot guarantee the formation of the normal interactions between the D and T-loops. The absence of these interactions would affect the juxtaposition of the two tRNA helical domains, potentially damaging the tRNA function. In addition to the short T-stem, this tRNA possesses another unprecedented feature, a very long D-stem consisting of seven base-pairs. Taken as such, a seven base-pair D-stem will also disrupt the normal interaction between the D and T-loops. On the other hand, the presence of the universal nucleotides in both the D and T-loops suggests that these loops probably interact with each other in the same way as in other tRNAs. Here, we demonstrate that the short T-stem and the long D-stem can naturally compensate each other, thus providing the normal D/T interactions. Molecular modeling has helped suggest a detailed scheme of mutual compensation between these two unique structural aspects of the archaeal selenocysteine tRNA. In the light of this analysis, other structural and functional characteristics of the selenocysteine tRNAs are discussed. PMID- 10390338 TI - Novel features in the structure of bovine ATP synthase. AB - The F1F0-ATP synthase from bovine heart mitochondria catalyses the synthesis of ATP from ADP and inorganic phosphate by using the energy of an electrochemical proton gradient derived from electron transport. The enzyme consists of three major domains: the globular F1catalytic domain of known atomic structure lies outside the lipid bilayer and is attached by a central stalk to the intrinsic membrane domain, F0, which transports protons through the membrane. Proton transport through F0evokes structural changes that are probably transmitted by rotation of the stalk to the catalytic sites in F1. In an alpha3beta3gamma1subcomplex, the rotation of the central gamma subunit driven by ATP hydrolysis has been visualised by optical microscopy. In order to prevent the alpha3beta3structure from following the rotation of the central gamma subunit, it has been proposed that the enzyme might have a stator connecting static parts in F0to alpha3beta3,thereby keeping it fixed relative to the rotating parts. Here we present electron microscopy images that reveal three new features in bovine F1F0 ATPase, one of which could be a stator. The second feature is a collar structure above the membrane domain and the third feature is some additional density on top of the F1domain. PMID- 10390337 TI - Internal correlations in minor groove profiles of experimental and computed B-DNA conformations. AB - It has been noticed that converged conformations of B-DNA oligomers obtained in MD calculations often have very small atom position rmsd values from the canonical B-DNA and all helical parameters close to the standard values, but their minor grooves tend to be somewhat narrower. This apparent bias disappears, however, when C5' rather than phosphorus atoms are used for measuring the groove width. At the origin of this effect is the specific orientation of phosphate groups in the canonical B-DNA model which maximizes their separation across the minor groove. When measured by C5' traces, minor groove profiles of experimental structures available in the Nucleic Acids Database show much less tendency to narrow below the canonical width. Correlation analysis reveals a high degree of correspondence in shapes of minor grooves of calculated and experimental single crystal structures of B-DNA oligomers. PMID- 10390339 TI - A set of plasmids constitutively producing different RNA levels in Escherichia coli. AB - New plasmids were developed for the in vivo expression of RNA in Escherichia coli. These plasmids combine constitutive promoters of different strengths with different origins of replication to provide a 75-fold range of expression of amber suppressor tRNA. The plasmids are either pMB1, p15A or temperature sensitive SC101 replicons, and can be used in two plasmid systems for studying RNA-protein interactions. The temperature-sensitive SC101 plasmids may be useful as gene replacement vectors. Another vector that is suitable for generating lethal mutations was constructed in a plasmid containing a regulatable phage T7 promoter. PMID- 10390340 TI - The reliability of in vivo structure-function analysis of tRNA aminoacylation. AB - The G.U wobble base-pair in the acceptor helix of Escherichia coli tRNAAlais critical for aminoacylation by the alanine synthetase. Previous work by several groups probed the mechanism of enzyme recognition of G.U by a structure-function analysis of mutant tRNAs using either a cell assay (amber suppressor tRNA) or a test tube assay (phage T7 tRNA substrate and purified enzyme). However, the aminoacylation capacity of particular mutant tRNAs was about 10(4)-fold higher in the cell assay. This led us to scrutinize the cell assay to determine if any parameter exaggerates the extent of aminoacylation in mutants forming substantial amounts of alanyl-tRNAAla. In doing so, we have refined and developed experimental designs to analyze tRNA function. We examined the level of aminoacylation of amber suppressor tRNAAlawith respect to the method of isolating aminoacyl-tRNA, the rate of cell growth, the cellular levels of alanine synthetase and elongation factor TU (EF-Tu), the amount of tRNA and the characteristics of EF-Tu binding. Within the precision of our measurements, none of these parameters varied in a way that could significantly amplify cellular alanyl-tRNAAla. A key observation is that the extent of aminoacylation of tRNAAlawas independent of tRNAAlaconcentration over a 75-fold range. Therefore, the cellular assay of tRNAAlareflects the substrate quality of the molecule for formation of alanyl-tRNAAla. These experiments support the authenticity of the cellular assay and imply that a condition or factor present in the cell assay may be absent in the test tube assay. PMID- 10390341 TI - Characterization of structural features important for T7 RNAP elongation complex stability reveals competing complex conformations and a role for the non-template strand in RNA displacement. AB - We have characterized the roles of the phage T7 RNA polymerase (RNAP) thumb subdomain and the RNA binding activity of the N-terminal domain in elongation complex (EC) stability by evaluating how disrupting these structures affects the dissociation rates of halted ECs. Our results reveal distinct roles for these elements in EC stabilization. On supercoiled or partially single-stranded templates the enzyme with a deletion of the thumb subdomain is exceptionally unstable. However, on linear duplex templates the polymerase which has been proteolytically cleaved within the N-terminal domain is the most unstable. The differences in the effects of these RNAP modifications on the stability of ECs on the different templates appear to be due to differences in EC structure: on the linear duplex templates the RNA is properly displaced from the DNA, but on the supercoiled or partially single-stranded templates an extended RNA:DNA hybrid makes a larger contribution to the conformational state of the EC. The halted EC can therefore exist either in a conformation in which the RNA is displaced from the DNA and forms an interaction with the RNAP, or in a conformation in which a more extended RNA:DNA hybrid is present and the RNA:RNAP interaction is less extensive. The partitioning between these competing conformations appears to be a function of the energetics of template reannealing and the relative strengths of the RNA:RNAP interaction and the RNA:DNA hybrid. PMID- 10390342 TI - Role of metal ions in the hydrolysis reaction catalyzed by RNase P RNA from Bacillus subtilis. AB - Precursor tRNA (ptRNA) substrates carrying a single Rp or Sp-phosphorothioate modification at the RNase P cleavage site were used as tools to study the cleavage mechanism of RNase P RNA from Bacillus subtilis. Both the Sp and the Rp diastereomer reduced the rate of processing at least 10(4)-fold under conditions where the chemical step is essentially rate-limiting. Neither the Rp nor the Sp phosphorothioate modification affected ptRNA ground state binding to B. subtilis RNase P RNA. Processing of the Rp-diastereomeric ptRNA could be restored in the presence of Mn2+or Cd2+, demonstrating direct metal ion coordination to the pro Rp oxygen during catalysis. With Cd2+, processing required the presence of another metal ion, such as Ca2+or Mg2+, to mediate substrate binding. This is in contrast to Escherichia coli RNase P RNA, which promotes cleavage of Rp diastereomeric ptRNA in the presence of Cd2+as the sole divalent metal ion. Analysis of [Cd2+]-dependent processing of the Rp-diastereomeric substrate by B. subtilis RNase P RNA was consistent with the involvement of at least two metal ions in catalysis. The presence of two catalytic metal ion binding sites is also supported by the inhibition mode of Ca2+on cleavage of unmodified ptRNA. In the presence of an Sp-phosphorothioate modification at the scissile bond, neither Mn2+nor Cd2+were able to restore significant cleavage at this location. Instead, the ribozyme promotes cleavage at the neighboring unmodified phosphodiester with low efficiency. Unaffected ground state binding of the Sp-diastereomeric ptRNA but a >/=10(4)-fold reduced hydrolysis rate may indicate a crucial role of the pro -Sp oxygen in transition state stabilization or may be attributed to steric exclusion of catalytic metal ions. Based on our comparative analyses of B. subtilis and E. coli RNase P RNA, each representing the main structural subtypes of bacterial RNase P RNA, common features in terms of active site constraints and role of catalytic metal ions can now be formulated for bacterial RNase P RNAs. On the other hand, substantial and unexpected differences with respect to the overall metal ion requirements and tRNA binding modes have been observed for the two catalytic RNAs. PMID- 10390343 TI - The role of Mg2+ and specific amino acid residues in the catalytic reaction of the major human abasic endonuclease: new insights from EDTA-resistant incision of acyclic abasic site analogs and site-directed mutagenesis. AB - Ape1, the major protein responsible for excising apurinic/apyrimidinic (AP) sites from DNA, cleaves 5' to natural AP sites via a hydrolytic reaction involving Mg2+. We report here that while Ape1 incision of the AP site analog tetrahydrofuran (F-DNA) was approximately 7300-fold reduced in 4 mM EDTA relative to Mg2+, cleavage of ethane (E-DNA) and propane (P-DNA) acyclic abasic site analogs was only 20 and 30-fold lower, respectively, in EDTA compared to Mg2+. This finding suggests that the primary role of the metal ion is to promote a conformational change in the ring-containing abasic DNA, priming it for enzyme mediated hydrolysis. Mutating the proposed metal-coordinating residue E96 to A or Q resulted in a approximately 600-fold reduced incision activity for both P and F DNA in Mg2+compared to wild-type. These mutants, while retaining full binding activity for acyclic P-DNA, were unable to incise this substrate in EDTA, pointing to an alternative or an additional function for E96 besides Mg2+ coordination. Other residues proposed to be involved in metal coordination were mutated (D70A, D70R, D308A and D308S), but displayed a relatively minor loss of incision activity for F and P-DNA in Mg2+, indicating a non-essential function for these amino acid residues. Mutations at Y171 resulted in a 5000-fold reduced incision activity. A Y171H mutant was fourfold less active than a Y171F mutant, providing evidence that Y171 does not operate as the proton donor in catalysis and that the additional role of E96 may be in establishing the appropriate active site environment via a hydrogen-bonding network involving Y171. D210A and D210N mutant proteins exhibited a approximately 25,000-fold reduced incision activity, indicating a critical role for this residue in the catalytic reaction. A D210H mutant was 15 to 20-fold more active than the mutants D210A or D210N, establishing that D210 likely operates as the leaving group proton donor. PMID- 10390344 TI - 3D imaging of the 58 kDa cell binding subunit of the Helicobacter pylori cytotoxin. AB - Pathogenic strains of Helicobacter pylori produce a potent exotoxin, VacA, which intoxicates gastric epithelial cells and leads to peptic ulcer. The toxin is released from the bacteria as a high molecular mass homo-oligomer of a 95 kDa polypeptide which undergoes specific proteolytic cleavage to 37 kDa and 58 kDa subunits. We have engineered a strain of H. pylori to delete the gene sequence coding for the 37 kDa subunit. The remaining 58 kDa subunit is expressed efficiently and exported as a soluble dimer that is non-toxic but binds target cells in a manner similar to the holotoxin. A 3D reconstruction of the molecule from electron micrographs of quick-freeze, deep-etched preparations reveals the contribution of each building block to the structure and permits the reconstruction of the oligomeric holotoxin starting from individual subunits. In this model P58 subunits are assembled in a ring structure with P37 subunits laying on the top. The data indicate that the 58 kDa subunit is capable of folding autonomously into a discrete structure recognizable within the holotoxin and containing the cell binding domain. PMID- 10390345 TI - Guiding a docking mode by phage display: selection of correlated mutations at the staphylokinase-plasmin interface. AB - During co-evolution of interacting proteins, functionally disruptive mutations on one side of the interface may be compensated by local amino acid changes on the other to restore binding affinity. This information can be useful for geometry based docking approaches by reducing the translational and rotational space available to the proteins. Here, we demonstrate that correlated mutations at a protein-protein interface can be rapidly identified by selecting a phage displayed library of a randomly mutated component of the complex for complementation of mutations that decreased binding in the interacting partner. This approach was used to deduce the binding mode of staphylokinase (Sak), a 15.5 kDa "indirect" plasminogen activator on microplasmin (microPli), the 28 kDa serine protease domain of plasmin. Biopanning indicated that residues Arg94 and Gly174 in microPli are located in close proximity to Glu75 and the Glu88:Ile128 pair in Sak, respectively. The coupled mutations Glu94<-->Lys75 reversed and Gly174<-->Lys88:Val128 introduced a salt bridge, whereby the binding affinities (with coupling energies of 1.8 to 2.3 kcal mol-1, respectively) and the plasminogen activation ability of the mutated complexes were partially restored. These findings suggested a unique docking mode of Sak at the western rim of the active-site cleft of microPli, that is in agreement with the structure of the Sak microPli complex as recently derived by other methods. PMID- 10390346 TI - Spermidine-induced aggregation of nucleosome core particles: evidence for multiple liquid crystalline phases. AB - We investigate the effect of the addition of a trivalent cation, spermidine, to dilute solutions of nucleosome core particles (NCP). In the presence of spermidine, part of the NCP segregates from the initial homogeneous solution, forming dense aggregates. We follow this precipitation process over a wide range of spermidine and NaCl concentrations and determine the conditions of aggregation of the particles. The structure of the dense phases is analyzed by means of polarizing light microscopy and cryo-electron microscopy. We report the existence of multiple supramolecular organizations. According to the relative concentrations of spermidine, monovalent salt and NCP, the particles may aggregate into amorphous phases, stack into randomly oriented columns, or form liquid crystalline phases. Two discotic liquid crystalline phases are identified and analyzed: a columnar nematic corresponding to columns of NCP simply aligned in parallel, and a columnar hexagonal phase in which the columns order into a transversal 2D hexagonal array. We discuss the nature and origin of the interactions possibly involved in the formation and maintenance of these different types of order. PMID- 10390347 TI - The N-terminal domain of the human Rad51 protein binds DNA: structure and a DNA binding surface as revealed by NMR. AB - Human Rad51 protein (HsRad51) is a homolog of Escherichia coli RecA protein, and functions in DNA repair and recombination. In higher eukaryotes, Rad51 protein is essential for cell viability. The N-terminal region of HsRad51 is highly conserved among eukaryotic Rad51 proteins but is absent from RecA, suggesting a Rad51-specific function for this region. Here, we have determined the structure of the N-terminal part of HsRad51 by NMR spectroscopy. The N-terminal region forms a compact domain consisting of five short helices, which shares structural similarity with a domain of endonuclease III, a DNA repair enzyme of E. coli. NMR experiments did not support the involvement of the N-terminal domain in HsRad51 HsBrca2 interaction or the self-association of HsRad51 as proposed by previous studies. However, NMR tiration experiments demonstrated a physical interaction of the domain with DNA, and allowed mapping of the DNA binding surface. Mutation analysis showed that the DNA binding surface is essential for double-stranded and single-stranded DNA binding of HsRad51. Our results suggest the presence of a DNA binding site on the outside surface of the HsRad51 filament and provide a possible explanation for the regulation of DNA binding by phosphorylation within the N-terminal domain. PMID- 10390348 TI - Crystal structure of alginate lyase A1-III from Sphingomonas species A1 at 1.78 A resolution. AB - The three-dimensional structure of alginate lyase A1-III (ALYIII) from a Sphingomonas species A1 was determined by X-ray crystallography. The enzyme was crystallized by the hanging-drop vapour-diffusion method in the presence of 49% ammonium sulfate at 20 degrees C. The crystals are monoclinic and belong to the space group C2 with unit cell dimensions of a=49.18 A, b=93.08 A, c=82.10 A and beta=104.12 degrees. There was one molecule of alginate lyase in the asymmetric unit of the crystal. The diffraction data up to 1. 71 A were collected with Rsymof 5.0%. The crystal structure of ALYIII was solved by the multiple isomorphous replacement method and refined at 1.78 A resolution using X-PLOR with a final R -factor of 18.0% for 10.0 to 1.78 A resolution data. The refined model of ALYIII contained 351 amino acid residues, 299 water molecules and two sulfate ions. The three-dimensional structure of ALYIII was abundant in helices and had a deep tunnel-like cleft in a novel (alpha6/alpha5)-barrel structure, which was similar to the (alpha6/alpha6)-barrel found in glucoamylase and cellulase. This structure presented the possibility that alginate molecules might penetrate into the cleft to interact with the catalytic site of ALYIII. PMID- 10390349 TI - Modulation of ligand binding in engineered human hemoglobin distal pocket. AB - Functional and structural studies on hemoglobin and myoglobin from different animals and engineered variants have enlightened the great importance of the physico-chemical properties of the side-chains at topological position B10 and E7. These residues proved to be crucial to the discrimination and stabilisation of gaseous ligands. In view of the data obtained on the high oxygen affinity hemoglobin from Ascaris worms and a new mutant of sperm whale myoglobin, we selected the two mutations Leu B10-->Tyr and His E7-->Gln as potentially relevant to control ligand binding parameters in the alpha and beta-chains of human hemoglobin. Here, we present an investigation of three new mutants: HbalphaYQ (alpha2YQbeta2A), HbbetaYQ (alpha2Abeta2YQ) and HbalphabetaYQ (alpha2YQbeta2YQ). They are characterised by a very low reactivity for NO, O2 and CO, and a reduced cooperativity. Their functional properties are not inconsistent with the behaviour expected for a two-state allosteric model. Proteins with these substitutions may be considered as candidates for the synthesis of a possible "blood substitute", which should yield an O2 adduct stable to autoxidation and slowly reacting with NO. The mutant HbalphabetaYQ is particularly interesting because the rate of reaction of NO with the oxy and deoxy derivatives is reduced. A structural interpretation of our data is presented based on the 3D structure of deoxy HbalphabetaYQ determined by crystallography at 1.8 A resolution. PMID- 10390350 TI - High-resolution structure of a potent, cyclic proteinase inhibitor from sunflower seeds. AB - Proteinaceous serine proteinase inhibitors are widespread throughout the plant kingdom where they play an important role in protection against pests and pathogens. Here, we describe the isolation and characterisation of a novel 14 amino acid residue cyclic peptide from sunflower seeds, which is a potent inhibitor of trypsin (Ki=100 pM). The crystal structure of this peptide in complex with bovine beta-trypsin shows both sequence and conformational similarity with the trypsin-reactive loop of the Bowman-Birk family of serine proteinase inhibitors. This inhibitor, however, is unique in being monofunctional, cyclic and far shorter (14 amino acid residues) than inhibitors belonging to this family (typically 60-70 amino acid residues). The high potency of this peptide is likely to arise from the considerable structural rigidity achieved through its cyclic nature which is further stabilised by a single internal disulphide bond. This study helps delineate the minimal unit required for effective peptide inhibitors of serine proteinases, and will assist in the further design of inhibitors to this widespread class of enzymes. PMID- 10390351 TI - Removal of the conserved disulfide bridges from the scFv fragment of an antibody: effects on folding kinetics and aggregation. AB - Fluorescence measurements and H/2H exchange experiments monitored by mass spectrometry have been applied to investigate the influence of the conserved disulfide bridges on the folding behavior and in vitro aggregation properties of the scFv fragment of the antibody hu4D5-8. A set of four proteins, carrying none, one, or both of the disulfide bridges have been compared regarding their stabilities, folding kinetics and tendency to aggregate. The results show that refolding of all four scFvs is ultimately limited by a slow proline isomerization in the VLdomain, since the native cis -conformation of proline L95 seems to be a prerequisite for formation of the native interface. Starting from short-term denatured protein, with the proline residues in their native conformation, a kinetically trapped intermediate is populated depending on the conditions, whose rate of conversion is slower than that of the fast-folding molecules. According to deuteron protection patterns determined by mass spectrometry, those domains retaining the disulfide bridge are able to form stable native-like structure, independent of native interface formation. The disulfide-free domains, in contrast, require the native interface for sufficient stabilization. The resistance of the scFvs towards aggregation seems to be critically dependent on the presence of the disulfide bridge in the VHdomain, and thus on the ability of the VHdomain to form stable structure prior to interaction with the VLdomain. The presence of a stable VLdomain in combination with a disulfide-free VHdomain appears to further promote aggregation, indicating the involvement of structured domains in the aggregates. PMID- 10390352 TI - Coiled-coil structure-mediated dimerization of template activating factor-I is critical for its chromatin remodeling activity. AB - Template activating factor-I (TAF-I)alpha and TAF-Ibeta have been identified as the host factors that activate DNA replication of the adenovirus genome complexed with viral basic core proteins (Ad core). TAF-I causes a structural change of the Ad core, thereby stimulating not only replication but also transcription from the Ad core DNA in vitro. TAF-I also activates transcription from the reconstituted chromatin consisting of DNA fragments and purified histones through chromatin remodeling. Although the carboxyl-terminal region, which is highly rich in acidic amino acids, is essential for the TAF-I activity, it remains unclear how other parts are involved in its activity. The native TAF-I isolated from HeLa cells exists as either hetero- or homo-oligomer. Here, we have demonstrated by cross linking assays that most of TAF-I exists as a dimer. Analyses using deletion mutant TAF-I proteins revealed that the amino-terminal region of TAF-I common to both alpha and beta is essential for dimerization. This region is predicted to form a coiled-coil structure. Indeed, mutations disrupting this putative structure abolished the dimerization capability and reduced the TAF-I activity in the Ad core DNA replication assay. Furthermore, we found that TAF-I mutants lacking the acidic tail act in a dominant-negative manner in this assay. These observations strongly suggest that the dimerization of TAF-I is important for its activity. PMID- 10390353 TI - Changing single side-chains can greatly enhance the resistance of a membrane protein to irreversible inactivation. AB - The thermal inactivation rates of a set of 20 cysteine-substituted variants of the integral membrane protein diacylglycerol kinase were measured. Two of the mutations, I53C and I70C, were found to significantly prolong the half-life of the enzyme in detergent solution. By combining the single mutants to create a double mutant, I53C/I70C, the half-life of the enzyme was improved from less than a minute at 70 degrees C to 51 minutes. These results demonstrate that individual side-chain substitutions can significantly improve the properties of membrane proteins in detergent solution. PMID- 10390354 TI - On the structure and operation of type I DNA restriction enzymes. AB - Type I DNA restriction enzymes are large, molecular machines possessing DNA methyltransferase, ATPase, DNA translocase and endonuclease activities. The ATPase, DNA translocase and endonuclease activities are specified by the restriction (R) subunit of the enzyme. We demonstrate that the R subunit of the Eco KI type I restriction enzyme comprises several different functional domains. An N-terminal domain contains an amino acid motif identical with that forming the catalytic site in simple restriction endonucleases, and changes within this motif lead to a loss of nuclease activity and abolish the restriction reaction. The central part of the R subunit contains amino acid sequences characteristic of DNA helicases. We demonstrate, using limited proteolysis of this subunit, that the helicase motifs are contained in two domains. Secondary structure prediction of these domains suggests a structure that is the same as the catalytic domains of DNA helicases of known structure. The C-terminal region of the R subunit can be removed by elastase treatment leaving a large fragment, stable in the presence of ATP, which can no longer bind to the other subunits of Eco KI suggesting that this domain is required for protein assembly. Considering these results and previous models of the methyltransferase part of these enzymes, a structural and operational model of a type I DNA restriction enzyme is presented. PMID- 10390355 TI - Modularity and homology: modelling of the type II module family from titin. AB - We report the homology modelling of the structures of the 162 type II modules from the giant multi-domain protein titin (also known as connectin). The package MODELLER was used and implemented in an automated fashion using four experimentally determined structures as templates. Validation of the models was assessed in terms of divergence from the templates and consensus of the alignments. The homology within the whole family of type II modules as well as with the templates is relatively high (20-35% identity and ca 50% similarity). Comparison between the models of domains for which an NMR structure has been solved and the experimental solution gives an estimate of the quality of the modelling. Our results allow us to distinguish between a set of structurally relevant residues, which are conserved throughout the whole family and buried in the hydrophobic core, from the residues that are conserved and exposed. These latter residues are potentially functionally important. Comparison of exposed conserved patches for modules in different regions of the titin molecule suggests potential interaction surfaces. Our results may be tested directly for those modules whose binding partner is known. PMID- 10390356 TI - Transproteomic evidence of a loop-deletion mechanism for enhancing protein thermostability. AB - Understanding the molecular determinants of protein thermostability is of theoretical and practical importance. While numerous determinants have been suggested, no molecular feature has been judged of paramount importance, with the possible exception of ion-pair networks. The difficulty in identifying the main determinants may have been the limited structural information available on the thermostable proteins. Recently the complete genomes for mesophilic, thermophilic and hyperthermophilic organisms have been sequenced, vastly improving the potential for uncovering general trends in sequence and structure evolution related to thermostability and, thus, for isolating the more important determinants. From a comparative analysis of 20 complete genomes, we find a trend towards shortened thermophilic proteins relative to their mesophilic homologs. Moreover, sequence alignments to proteins of known structure indicate that thermophilic sequences are more likely than their mesophilic homologs to have deletions in exposed loop regions. The new genomes offer enough comparable sequences to compute meaningful statistics that point to loop deletion as a general evolutionary strategy for increasing thermostability. PMID- 10390357 TI - The cavity and pore helices in the KcsA K+ channel: electrostatic stabilization of monovalent cations. AB - The electrostatic influence of the central cavity and pore alpha helices in the potassium ion channel from Streptomyces lividans (KcsA K+ channel) was analyzed by solving the finite difference Poisson equation. The cavity and helices overcome the destabilizing influence of the membrane and stabilize a cation at the membrane center. The electrostatic effect of the pore helices is large compared to that described for water-soluble proteins because of the low dielectric membrane environment. The combined contributions of the ion self energy and the helix electrostatic field give rise to selectivity for monovalent cations in the water-filled cavity. Thus, the K+ channel uses simple electrostatic principles to solve the fundamental problem of ion destabilization by the cell membrane lipid bilayer. PMID- 10390358 TI - Paracellin-1, a renal tight junction protein required for paracellular Mg2+ resorption. AB - Epithelia permit selective and regulated flux from apical to basolateral surfaces by transcellular passage through cells or paracellular flux between cells. Tight junctions constitute the barrier to paracellular conductance; however, little is known about the specific molecules that mediate paracellular permeabilities. Renal magnesium ion (Mg2+) resorption occurs predominantly through a paracellular conductance in the thick ascending limb of Henle (TAL). Here, positional cloning has identified a human gene, paracellin-1 (PCLN-1), mutations in which cause renal Mg2+ wasting. PCLN-1 is located in tight junctions of the TAL and is related to the claudin family of tight junction proteins. These findings provide insight into Mg2+ homeostasis, demonstrate the role of a tight junction protein in human disease, and identify an essential component of a selective paracellular conductance. PMID- 10390359 TI - Inhibition of the interferon-inducible protein kinase PKR by HCV E2 protein. AB - Most isolates of hepatitis C virus (HCV) infections are resistant to interferon, the only available therapy, but the mechanism underlying this resistance has not been defined. Here it is shown that the HCV envelope protein E2 contains a sequence identical with phosphorylation sites of the interferon-inducible protein kinase PKR and the translation initiation factor eIF2alpha, a target of PKR. E2 inhibited the kinase activity of PKR and blocked its inhibitory effect on protein synthesis and cell growth. This interaction of E2 and PKR may be one mechanism by which HCV circumvents the antiviral effect of interferon. PMID- 10390360 TI - Replication of subgenomic hepatitis C virus RNAs in a hepatoma cell line. AB - An estimated 170 million persons worldwide are infected with hepatitis C virus (HCV), a major cause of chronic liver disease. Despite increasing knowledge of genome structure and individual viral proteins, studies on virus replication and pathogenesis have been hampered by the lack of reliable and efficient cell culture systems. A full-length consensus genome was cloned from viral RNA isolated from an infected human liver and used to construct subgenomic selectable replicons. Upon transfection into a human hepatoma cell line, these RNAs were found to replicate to high levels, permitting metabolic radiolabeling of viral RNA and proteins. This work defines the structure of HCV replicons functional in cell culture and provides the basis for a long-sought cellular system that should allow detailed molecular studies of HCV and the development of antiviral drugs. PMID- 10390361 TI - Positive selection of natural autoreactive B cells. AB - Lymphocyte development is critically influenced by self-antigens. T cells are subject to both positive and negative selection, depending on their degree of self-reactivity. Although B cells are subject to negative selection, it has been difficult to test whether self-antigen plays any positive role in B cell development. A murine model system of naturally generated autoreactive B cells with a germ line gene-encoded specificity for the Thy-1 (CD90) glycoprotein was developed, in which the presence of self-antigen promotes B cell accumulation and serum autoantibody secretion. Thus, B cells can be subject to positive selection, generated, and maintained on the basis of their autoreactivity. PMID- 10390362 TI - Protein sorting by directed maturation of Golgi compartments. AB - How does the Golgi stack mediate transport of cargo from the endoplasmic reticulum (ER) to the cell surface? A possibility is that cargo-containing vesicles derived from the ER form early Golgi compartments that then mature by retrieval of processing enzymes from later Golgi compartments. Maturation continues at terminal Golgi compartments by retrieval of transport components from the endocytic pathway to promote sorting of cargo to multiple cellular destinations. Hence, retrograde movement may integrate exocytic and endocytic pathways in eukaryotic cells and coordinate membrane flow and cargo transport through the Golgi stack. PMID- 10390363 TI - Structural rearrangements underlying K+-channel activation gating. AB - The intramembrane molecular events underlying activation gating in the Streptomyces K+ channel were investigated by site-directed spin-labeling methods and electron paramagnetic resonance spectroscopy. A comparison of the closed and open conformations of the channel revealed periodic changes in spin-label mobility and intersubunit spin-spin interaction consistent with rigid-body movements of the two transmembrane helices TM1 and TM2. These changes involve translations and counterclockwise rotations of both helices relative to the center of symmetry of the channel. The movement of TM2 increases the diameter of the permeation pathway along the point of convergence of the four subunits, thus opening the pore. Although the extracellular residues flanking the selectivity filter remained immobile during gating, small movements were detected at the C terminal end of the pore helix, with possible implications to the gating mechanism. PMID- 10390364 TI - SHRIMP uranium-lead dating of diagenetic xenotime in siliciclastic sedimentary rocks AB - Diagenetic xenotime is common in siliciclastic sedimentary rocks, where it starts to form on detrital zircon shortly after sediment deposition. It is possible to estimate the age of sedimentary rocks by in situ uranium-lead analysis of that xenotime. Two Proterozoic sandstone units from northwestern Australia, previously constrained to the age interval of 1790 to 750 million years ago, have diagenetic xenotime ages of 1704 +/- 7 and 1704 +/- 14 million years ago. This method has potential for dating sedimentary sequences of all ages but should be especially valuable for refining the Precambrian time scale. PMID- 10390365 TI - Experimental detection of hydrogen trioxide AB - Hydrogen trioxide (HO3) has long been postulated as a key intermediate in important atmospheric processes but has proved difficult to detect. The molecule was unequivocally detected in experiments based on neutralization-reionization and neutralization-reionization/collisionally activated dissociation mass spectrometry, using protonated ozone (HO3+) as the charged precursor. Hydrogen trioxide is a relatively stable species and has a H-O-O-O connectivity and a lifetime exceeding 10(-6) seconds at ambient temperature. PMID- 10390366 TI - Microfabrication inside capillaries using multiphase laminar flow patterning AB - The reaction of species in solutions flowing laminarly (without turbulent mixing) inside capillaries was used as the basis for a broadly applicable method of microfabrication. In this method, patterning occurs as a result of transport of reactive species to interfaces within the capillary by laminar flow. A wide range of chemistries can be used to generate structures with feature sizes of less than 5 micrometers and with spatial localization to within 5 micrometers. The method is applicable to the patterning of metals, organic polymers, inorganic crystals, and ceramics on the inner walls of preformed capillaries, using both additive and subtractive processes. PMID- 10390367 TI - Shock melting of the canyon diablo impactor: constraints from nickel-59 contents and numerical modeling AB - Two main types of material survive from the Canyon Diablo impactor, which produced Meteor Crater in Arizona: iron meteorites, which did not melt during the impact; and spheroids, which did. Ultrasensitive measurements using accelerator mass spectrometry show that the meteorites contain about seven times as much nickel-59 as the spheroids. Lower average nickel-59 contents in the spheroids indicate that they typically came from 0.5 to 1 meter deeper in the impactor than did the meteorites. Numerical modeling for an impact velocity of 20 kilometers per second shows that a shell 1.5 to 2 meters thick, corresponding to 16 percent of the projectile volume, remained solid on the rear surface; that most of the projectile melted; and that little, if any, vaporized. PMID- 10390368 TI - Atomic-scale quasi-particle scattering resonances in Bi2Sr2CaCu2O8+delta AB - Low-temperature scanning tunneling spectroscopy of the high transition temperature (high-Tc) cuprate Bi2Sr2CaCu2O8+delta reveals the existence of large numbers of identical regions with diameters of about 3 nanometers that have a relatively high density of low-energy quasi-particle states. Their spatial and spectroscopic characteristics are consistent with theories of strong quasi particle scattering from atomic-scale impurities in a d-wave superconductor. These characteristics include breaking of local particle-hole symmetry, a diameter near twice the superconducting coherence length, and an inverse square dependence of their local density-of-states on distance from the scattering center. In addition to the validation of d-wave quasi-particle scattering theories, these observations identify a source for the anomalously high levels of low-energy quasi-particles in Bi2Sr2CaCu2O8+delta at low temperatures. PMID- 10390369 TI - High H2 uptake by alkali-doped carbon nanotubes under ambient pressure and moderate temperatures AB - Lithium- or potassium-doped carbon nanotubes can absorb approximately 20 or approximately 14 weight percent of hydrogen at moderate (200 degrees to 400 degrees C) or room temperatures, respectively, under ambient pressure. These values are greater than those of metal hydride and cryoadsorption systems. The hydrogen stored in the lithium- or potassium-doped carbon nanotubes can be released at higher temperatures, and the sorption-desorption cycle can be repeated with little decrease in the sorption capacity. The high hydrogen-uptake capacity of these systems may be derived from the special open-edged, layered structure of the carbon nanotubes made from methane, as well as the catalytic effect of alkali metals. PMID- 10390370 TI - Regulation of NMDA receptors by an associated phosphatase-kinase signaling complex. AB - Regulation of N-methyl-D-aspartate (NMDA) receptor activity by kinases and phosphatases contributes to the modulation of synaptic transmission. Targeting of these enzymes near the substrate is proposed to enhance phosphorylation-dependent modulation. Yotiao, an NMDA receptor-associated protein, bound the type I protein phosphatase (PP1) and the adenosine 3',5'-monophosphate (cAMP)-dependent protein kinase (PKA) holoenzyme. Anchored PP1 was active, limiting channel activity, whereas PKA activation overcame constitutive PP1 activity and conferred rapid enhancement of NMDA receptor currents. Hence, yotiao is a scaffold protein that physically attaches PP1 and PKA to NMDA receptors to regulate channel activity. PMID- 10390371 TI - A phospholipase C-dependent inositol polyphosphate kinase pathway required for efficient messenger RNA export. AB - In order to identify additional factors required for nuclear export of messenger RNA, a genetic screen was conducted with a yeast mutant deficient in a factor Gle1p, which associates with the nuclear pore complex (NPC). The three genes identified encode phospholipase C and two potential inositol polyphosphate kinases. Together, these constitute a signaling pathway from phosphatidylinositol 4, 5-bisphosphate to inositol hexakisphosphate (IP6). The common downstream effects of mutations in each component were deficiencies in IP6 synthesis and messenger RNA export, indicating a role for IP6 in GLE1 function and messenger RNA export. PMID- 10390372 TI - Reactive nitrogen species and tyrosine nitration in the respiratory tract: epiphenomena or a pathobiologic mechanism of disease? PMID- 10390373 TI - The impact of HIV infection on Mycobacterium kansasii disease in South African gold miners. AB - The impact of human immunodeficiency virus (HIV) infection on Mycobacterium kansasii disease in miners was investigated with a retrospective study covering a single workforce. M. kansasii, isolated from 43 HIV-positive and 202 HIV-negative miners, was the most common nontuberculous mycobacterial (NTM) species in both HIV groups. CD4 counts were unusually high for M. kansasii disease (mean 490 x 10(6)/L, from 14 HIV-positive men). Treatment outcomes were similar: mortality during treatment was higher in HIV-positive than in HIV-negative men (9% and 2%, respectively), but not significantly so. The majority of a sample of 31 HIV positive and 92 HIV-negative men had radiological silicosis and/or old tuberculosis scarring prior to M. kansasii disease. A normal premorbid radiograph was more common in HIV-positive men (45% versus 24%; odds ratio [OR], 2.62; 95% confidence interval [95% CI], 1.01 to 6.67). New cavitation was less common (55% versus 78%; OR, 0.34; 95% CI, 0.13 to 0.88) and new hilar adenopathy more common (OR, 5.07; 95% CI, 1.24 to 21.9) in HIV-positive than in HIV-negative men. Miners, who have additional NTM risk factors, develop M. kansasii disease that occurs at an earlier stage of HIV infection and more closely resembles disease in HIV-negative men than has been found for HIV-associated M. kansasii disease in other settings. PMID- 10390374 TI - Nontuberculous mycobacteria: defining disease in a prospective cohort of South African miners. AB - A gold mining work force was followed prospectively over 1 yr for sputum nontuberculous mycobacterial (NTM) isolates. NTM were isolated from 118 men, of whom 32 (27%) met the American Thoracic Society (ATS) case-definitions for pulmonary NTM disease (23 M. kansasii, seven M. scrofulaceum, one M. avium, and one M. abscessus). Determining isolate significance was difficult because most men had been started on presumptive TB treatment before isolate identification (70%). Histologic criteria were considered inappropriate for this high M. tuberculosis incidence population, particularly for patients who had stabilized on presumptive TB treatment. Among men not meeting case-definitions, indicators of disease were significantly more prevalent for M. kansasii than for M. fortuitum, the local laboratory contaminant (ORs: 6.5 for cough, 7. 2 for smear positivity, 36.0 for radiologic changes, and 14.3 for presumptive TB treatment), suggesting underdiagnosis of M. kansasii disease. Of 53 men with definite or possible M. kansasii disease, 18 (34%) were HIV-positive. HIV-associated M. kansasii disease occurred at an early stage of immunosuppression (median CD4 count, 381 x 10(6)/L) with a good outcome (83% cured after 12 mo of treatment). ATS case-definitions for NTM disease are difficult to apply in this population, and treatment decisions should be guided by the pathogenic potential of the isolate. PMID- 10390375 TI - Calibration of seven ICU ventilators for mechanical ventilation with helium oxygen mixtures. AB - The study evaluated seven intensive care unit (ICU) ventilators (Veolar FT, Galileo, Evita 2, Evita 4, Servo 900C, Servo 300, Nellcor Puritan Bennett 7200 Series) with helium-oxygen (HeO2), using a lung model, to develop correction factors for the safe use of HeO2. A 70:28 helium-O2 mixture (heliox) replaced air and combined with O2 (HeO2). Theoretical impact of HeO2 on inspiratory valves and gas mixing was computed. True fraction of inspired oxygen (FIO2del) was compared with fraction of inspired oxygen (FIO2) set on the ventilator (FIO2set). True tidal volume (VTdel) was compared with VT set on the ventilator (VTset) in volume control and with control VTdel at FIO2 1.0 in pressure control. FIO2del minimally exceeded FIO2set ( FIO2set by 125%). In volume control, with the Veolar FT, Galileo, Evita 2, and Servo 900C, VTdel > VTset, with the 7200 Series VTdel < VTset (linear relationship, magnitude of discrepancy inversely related to FIO2set). With the Evita 4, VTdel > VTset (nonlinear relationship), whereas with the Servo 300 VTdel = VTset. In pressure control, VTdel was identical to control measurements, except with the 7200 Series (ventilator malfunction). Correction factors were developed that can be applied to most ventilators. PMID- 10390376 TI - The effect of exposure to ozone and nitrogen dioxide on the airway response of atopic asthmatics to inhaled allergen: dose- and time-dependent effects. AB - Eleven mild atopic asthmatic patients were exposed for 6 h, in randomized order, to air, 100 ppb O3, 200 ppb NO2, and 100 ppb O3 + 200 ppb NO2, followed immediately by bronchial allergen challenge. Subsequently 10 of these patients were exposed for 3 h to air, 200 ppb O3, 400 ppb NO2, and 200 ppb O3 + 400 ppb NO2, followed immediately by bronchial allergen challenge. All exposures were carried out in an environmental chamber, with intermittent moderate exercise, and a minimal interval of 2 wk. Exposure for 6 h to 100 ppb O3, 200 ppb NO2, and 100 ppb O3 + 200 ppb NO2 did not lead to any significant increase in the airway response of these individuals to inhaled allergen, when compared with exposure for 6 h to air. In contrast, exposure for 3 h to 200 ppb O3, 400 ppb NO2, and 200 ppb O3 + 400 ppb NO2 significantly decreased the dose of allergen (in log cumulative breath units [CBU]) required to decrease FEV1 by 20% (allergen PD20FEV1), compared with exposure to air (geometric mean CBU: 3.0 for air versus 2.66 for O3 [p = 0.002]; 2.78 for NO2 [p = 0. 018]; 2.65 for O3 + NO2 [p = 0.002]). These results suggest that the pollutant-induced changes in airway response of mild atopic asthmatics to allergen may be dependent on a threshold concentration rather than the total amount of pollutant inhaled over a period of time. PMID- 10390377 TI - Markers of impaired growth of pulmonary function in children and adolescents. AB - Our knowledge about the age-related growth of pulmonary function is incomplete. The purpose of this study was to describe the relation of various factors to the growth of pulmonary function in children and adolescents. A population sample comprising 408 children and adolescents (7-17 yr of age at enrollment) was reexamined after a 6-yr interval. Case history was obtained by interview and questionnaire. Pulmonary function, skin prick test reactivity to common allergens, and airway responsiveness (AR) were measured using standard techniques; airway hyperresponsiveness (AHR) was defined as a concentration of histamine causing a 20% decline in FEV1 < 8 mg/ml. The cross-sectional analyses of data from the two surveys showed that the presence of asthma (p < 0.02), atopy to house dust mite (HDM) (p = 0.03), and increasing degree of AR (p < 0.002) were associated with a lower level of FEV1 %pred. The longitudinal analysis revealed that asthma (p = 0.0001) and a lower level of FEV1 (p < 0.0001) at enrollment were associated with a lower level of FEV1 at follow-up. Further, an increase in the degree of AR (p = 0. 0001), new asthma (p = 0.0002), and new atopy to HDM (p = 0.03) also predicted a lower level of FEV1 at the end of the observation period. Confining the analysis to subjects without asthma and without evidence of AHR (n = 271) showed that both persistent (p = 0.04) and new (p = 0.03) atopy to HDM predicted a lower level of FEV1 at follow-up; compared with subjects with a negative skin reaction to HDM, those subjects who were sensitized to HDM had on average a 5%pred lower level of FEV1. The growth of FEV1 in children and adolescents appears to be impaired not only by symptomatic asthma but also by an increase in the degree of AR and atopy to HDM; sensitization to HDM appears to have a negative impact on the age-related growth in FEV1 even in nonasthmatic subjects without evidence of AHR. PMID- 10390379 TI - Neuropsychological sequelae and impaired health status in survivors of severe acute respiratory distress syndrome. AB - Acute respiratory distress syndrome (ARDS) is a disease of acute respiratory failure manifested by severe hypoxemia with a high mortality rate. Previous outcome studies of ARDS have assessed survival and/or pulmonary function as the primary outcome variables. Cognitive or psychological outcomes following ARDS have not been described, despite the possibility that ARDS patients are at risk for brain injury through hypoxemia or other mechanisms. In the current study 55 consecutive ARDS survivors completed a battery of neuropsychological tests and questionnaires regarding health status, cognitive and psychological outcomes at the time of hospital discharge and 1 yr after onset of ARDS. At hospital discharge, 100% (55 of 55) of survivors exhibited cognitive and affective impairments, as well as problems with health status which affected their quality of life. At 1 yr after ARDS, 17 of 55 (30%) patients still exhibited generalized cognitive decline. Forty-three of 55 (78%) patients had all or at least one of the following: impaired memory, attention, concentration and/or decreased mental processing speed. One year after ARDS a substantial portion of ARDS survivors exhibit impaired health status and cognitive sequelae which may be due to hypoxemia, emboli, inflammation, drug toxicity, and/or other etiologies. PMID- 10390378 TI - Exhaled nitric oxide, sensitization, and exposure to allergens in patients with asthma who are not taking inhaled steroids. AB - The level of exhaled nitric oxide (eNO) is elevated in patients with asthma and eNO may be involved in airway inflammation. Exposure to allergen in sensitized individuals may contribute to airway inflammation. Our aim was to investigate the relationship between eNO, sensitization, and exposure to indoor allergen in nonsmoking adults with asthma who are not taking inhaled steroids. In subjects with a positive methacholine challenge (PD20 < 4 mg) we measured eNO (LR 2000 chemiluminesence analyzer); serum total and specific IgE; skin test to mite, cat, and dog; and allergen levels in domestic dust (Der p 1, Fel d 1, and Can f 1). Subjects were classified as exposed or not exposed to allergen according to previously proposed significant levels (> 2 micrograms/g Der p 1, > 8 micrograms/g Fel d 1, and > 10 micrograms/g Can f 1). Of the 43 subjects (> 95% atopic) complete data were available for 38, of whom 26 were both sensitized and exposed to one or more allergen and 12 were sensitized but not exposed to any allergens. eNO was significantly higher in those subjects who were both sensitized and exposed to indoor allergen than in those who were sensitized but not exposed (GM and 95% CI: 17. 69 [14.1- 22.15] versus 9.09 [6.5-12.7], p = 0.001). Levels of eNO are significantly higher in patients with asthma who are both sensitized and exposed to relevant allergen than in those who were sensitized but not exposed. eNO may be a marker of the airway inflammation induced by domestic exposure to allergen in sensitized patients with asthma. PMID- 10390380 TI - Skeletal muscle oxidative capacity, fiber type, and metabolites after lung transplantation. AB - Lung transplant (LTx) recipients have a low peak work rate, peak oxygen consumption (V O2peak), and early lactate threshold on incremental exercise. We hypothesized that LTx recipients have reduced oxidative function and altered fiber type proportion in peripheral skeletal muscle. Seven stable LTx recipients and seven age- and sex-matched control subjects were studied. Incremental exercise testing with arterialized venous sampling and a resting quadriceps femoris punch muscle biopsy were performed. Muscle specimens were analyzed for fiber type proportion, metabolites, oxidative and glycolytic enzyme activities, and mitochondrial ATP production rate (MAPR) using standard techniques. The results showed that mean V O2peak in LTx recipients was 52% of control subjects. Compared with the control subjects, LTx skeletal muscle exhibited: (1) a lower MAPR; (2) lower activity of the mitochondrial enzymes glutamate dehydrogenase (GDH), citrate synthase (CS), 2-oxogluterate dehydrogenase (OGDH), and 3 hydroxyacyl-CoA-dehydrogenase (HAD). There was no difference in the activities of anaerobic enzymes, except for higher phosphofructokinase activity; (3) a lower proportion of type I fibers; (4) a higher lactate and inosine monophosphate (IMP) content and a lower ATP content at rest indicating a high reliance on anaerobic metabolism. The reduced type I fiber proportion and severely reduced mitochondrial oxidative capacity may play an important role in exercise limitation after LTx. PMID- 10390381 TI - Collapsibility of passive pharynx in patients with acromegaly. AB - Sleep-disordered breathing (SDB), either central or obstructive in nature, is common in patients with acromegaly. However, no study has systematically examined the collapsibility of the pharynx in acromegaly to date. We evaluated intrinsic mechanical properties of passive pharynx in 10 anesthetized and paralyzed patients with active acromegaly before transsphenoidal adenomectomy for their pituitary adenoma. Static pressure-area relationships of the velopharynx and oropharynx were obtained by step changes in airway pressure during endoscopic cross-sectional area measurement of each segment. Moreover, curve fitting analysis by an exponential function estimated the closing pressure (P'close) of each segment. Preoperative nocturnal oximetry identified five acromegalic patients with an oxygen desaturation index (ODI) greater than 10 h-1 and clinical symptoms suggesting presence of SDB. The pharyngeal airway of all five acromegalic patients with SDB was highly collapsible at both velopharynx and oropharynx with positive P'close. Compared with age-, body mass index (BMI)-, and ODI-matched SDB patients without acromegaly, SDB patients with acromegaly had a higher P'close of the oropharynx, indicating that the etiology of SDB in acromegaly appears to differ from that of ordinary sleep apnea. Our results suggest that anatomic abnormality, especially at the base of the tongue, appears to play a significant role in development of SDB in acromegaly. PMID- 10390382 TI - Right heart catheterization in acute lung injury: an observational study. AB - Right heart catheterization (RHC) is commonly used in the diagnosis and management of acute lung injury (ALI). However, controversy exists regarding RHC. We examined RHC use during the first 3 d of ALI in an observational study of 135 patients defined by American-European Consensus Conference criteria. Study parameters examined for association with RHC included the Acute Physiology and Chronic Health Evaluation (APACHE) III score, lung injury score (LIS), and 20 additional epidemiologic, clinical, and laboratory parameters. RHC was performed in 70 patients (52%) within the first 3 d of ALI. RHC was positively associated (p < 0.05) with a diagnosis of sepsis, APACHE III score, blood urea nitrogen (BUN), creatinine, net fluid balance, and positive end-expiratory pressure. RHC was negatively associated (p < 0.05) with mean arterial pressure (Pa) and PaO2/FIO2. Logistic regression identified four predictors for RHC placement: sepsis, PaO2/FIO2, BUN, and Pa. Initial right atrial and pulmonary artery occlusion pressure measurements demonstrated a moderately strong correlation (r = 0.72). Use of RHC was associated with a change in one or more therapeutic interventions (intravascular fluids, vasopressors, diuretics) in 78% of patients. In summary, patients receiving RHC during the first 3 d of ALI were more severely ill than those who did not receive RHC, and RHC was associated with a change in therapy in most patients. PMID- 10390383 TI - Expiratory washout versus optimization of mechanical ventilation during permissive hypercapnia in patients with severe acute respiratory distress syndrome. AB - The aim of this study was to compare three ventilatory techniques for reducing PaCO2 in patients with severe acute respiratory distress syndrome treated with permissive hypercapnia: (1) expiratory washout alone at a flow of 15 L/min, (2) optimized mechanical ventilation defined as an increase in the respiratory frequency to the maximal rate possible without development of intrinsic positive end- expiratory pressure (PEEP) combined with a reduction of the instrumental dead space, and (3) the combination of both methods. Tidal volume was set according to the pressure-volume curve in order to obtain an inspiratory plateau airway pressure equal to the upper inflection point minus 2 cm H2O after setting the PEEP at 2 cm H2O above the lower inflection point and was kept constant throughout the study. The three modalities were compared at the same inspiratory plateau airway pressure through an adjustment of the extrinsic PEEP. During conventional mechanical ventilation using a respiratory frequency of 18 breaths/min, respiratory acidosis (PaCO2 = 84 +/- 24 mm Hg and pH = 7.21 +/- 0.12) was observed. Expiratory washout and optimized mechanical ventilation (respiratory frequency of 30 +/- 4 breaths/min) had similar effects on CO2 elimination (DeltaPaCO2 = -28 +/- 11% versus -27 +/- 12%). A further decrease in PaCO2 was observed when both methods were combined (DeltaPaCO2 = -46 +/- 7%). Extrinsic PEEP had to be reduced by 5.3 +/- 2.1 cm H2O during expiratory washout and by 7.3 +/- 1.3 cm H2O during the combination of the two modes, whereas it remained unchanged during optimized mechanical ventilation alone. In conclusion, increasing respiratory rate and reducing instrumental dead space during conventional mechanical ventilation is as efficient as expiratory washout to reduce PaCO2 in patients with severe ARDS and permissive hypercapnia. When used in combination, both techniques have additive effects and result in PaCO2 levels close to normal values. PMID- 10390384 TI - Noninvasive ventilation as a systematic extubation and weaning technique in acute on-chronic respiratory failure: a prospective, randomized controlled study. AB - Prolonged duration of endotracheal mechanical ventilation (ETMV) is associated with an increased morbidity and mortality in intensive care unit (ICU) patients. The aim of this study was to assess the usefulness of noninvasive ventilation (NIV) as a systematic extubation and weaning technique to reduce the duration of ETMV in acute-on-chronic respiratory failure (ACRF). Among 53 consecutively intubated patients admitted for ACRF, we conducted a prospective, randomized controlled trial of weaning in 33 patients who failed a 2-h T-piece weaning trial (2 h-WT) although they met simple criteria for weaning. Conventional invasive pressure support ventilation (IPSV) was used as the control weaning technique in 16 patients (IPSV group), and NIV was applied immediately after extubation in 17 patients (NIV group). The two weaning groups were similar for type of chronic respiratory failure (CRF), pulmonary function data, age, Simplified Acute Physiology Score (SAPS II), and severity of ACRF on admission. The characteristics of the two groups were also similar at randomization. In the IPSV group, 12 of 16 patients (75%) were successfully weaned and extubated, versus 13 of 17 (76.5%) in the NIV group (p = NS). NIV like IPSV significantly and similarly improved gas exchange in relation to that achieved during 2 h-WT (p < 0.05). The duration of ETMV was significantly shorter in the NIV (4.56 +/- 1.85 d) than in the IPSV group (7.69 +/- 3.79 d) (p = 0. 004). NIV also reduced the mean period of daily ventilatory support, but increased the total duration of ventilatory support related to weaning (3.46 +/- 1.42 d, versus 11.54 +/- 5.24 d with NIV; p = 0. 0001). Most patients in the IPSV group developed complications related to ETMV and/or the weaning process, but the difference was not significant (nine of 16 versus six of 17). The durations of ICU and hospital stays and the 3-mo survival were similar in the two groups. In conclusion, NIV permits earlier removal of the endotracheal tube than with conventional IPSV, and reduces the duration of daily ventilatory support without increasing the risk of weaning failures. NIV should be considered as a new and useful systematic approach to weaning in patients with ACRF who are difficult to wean. PMID- 10390385 TI - Lung function and sputum characteristics of patients with severe asthma during an induced exacerbation by double-blind steroid withdrawal. AB - Some patients with severe asthma are difficult to control and suffer from frequent exacerbations, whereas others remain stable with anti-inflammatory therapy. To investigate mechanisms of exacerbations, we compared 13 patients 20 to 51 yr of age (11 female, two male) with difficult-to-control asthma (two or more exacerbations during the previous year) and 15 patients 20 to 47 yr of age (13 female, two male) with severe but stable asthma (no exacerbations) after matching for sex, age, atopy, lung function, airway responsiveness, and medication. Exacerbations were induced by double-blind, controlled tapering of inhaled corticosteroids (fluticasone propionate) at weekly intervals. FEV1, airway responsiveness for methacholine (PC20MCh) and hypertonic saline (HYP slope), eosinophils and soluble markers (ECP, albumin, IL-6, IL-8) in induced sputum were assessed at baseline and during exacerbation (peak flow < 60% of personal best), or after 5 wk if no exacerbation occurred. Steroid tapering caused a decrease (mean +/- SEM) in FEV1 (12.1 +/- 3.1% pred; p = 0.045), PC20MCh (2.1 +/- 0.4 doubling dose; p = 0.004) and HYP slope (1.7 +/- 0.3 doubling dose; p = 0.001), and an increase in sputum eosinophils (10 +/- 3%; p = 0.008) and soluble markers for the two groups combined, without significant differences between the groups. Patients with difficult-to-control asthma had more exacerbations than did the stable asthmatics during both steroid tapering (7 versus 2; p = 0.022) and corticosteroid treatment (6 versus 0; p = 0.003). Exacerbations during steroid treatment in the patients with difficult-to-control asthma were associated with a decrease in FEV1 and PC20MCh, but not in HYP slope or increase in sputum eosinophils. We conclude that tapering of inhaled corticosteroids induces a rapid, reversible flare-up of eosinophilic airway inflammation. Patients with difficult-to-control asthma may develop exacerbations despite treatment with inhaled corticosteroids, which appear to have an eosinophil-independent mechanism. This implies that assessment of the nature of exacerbations may contribute to improved treatment for these patients. PMID- 10390386 TI - Rhinovirus-16 colds in healthy and in asthmatic subjects: similar changes in upper and lower airways. AB - Rhinovirus (RV) infections appear to precipitate most asthma exacerbations. To investigate whether RV-16 induces different inflammatory changes in upper and lower airways of asthmatic and healthy subjects, we inoculated 10 nonatopic healthy and 11 atopic asthmatic adults with 2,000 TCID50 RV-16. Subjects recorded symptoms and peak flow daily; and they underwent spirometry, methacholine challenge (PC20), nasal lavage, and sputum induction at baseline and on Days 2, 4, 15, and 29 d after inoculation. One asthmatic subject developed an exacerbation requiring prednisone treatment 5 d after inoculation. The cold symptom severity (Jackson score) did not differ between groups. During the cold, asthma symptoms increased slightly from baseline in the asthmatic group; and PC20 decreased in the healthy group. However, peak flow, bronchodilator use, and spirometry did not change in either group. At baseline, asthmatics had higher neutrophils, eosinophils, and interleukin (IL)-6 in nasal lavage. After inoculation, both groups developed significant increases in nasal neutrophils, IL 6 and IL-8, and modest increases in sputum neutrophils and IL-6, but not IL-8. However, these changes did not differ between groups. IL-5, interferon-gamma, and RANTES were detected only in nasal lavages from two asthmatic subjects, who had the most severe colds. IL-11 was not detected in any sample. We conclude that inflammatory responses of upper and lower airways during RV-16 colds are similar in asthmatic and healthy subjects, and that RV-16 infection is not by itself sufficient to provoke clinical worsening of asthma. PMID- 10390387 TI - Mechanical ventilation affects local and systemic cytokines in an animal model of acute respiratory distress syndrome. AB - We examined the hypothesis that injurious ventilatory strategies (large tidal volume [VT] and/or low positive end-expiratory pressure [PEEP]) would increase release of inflammatory mediators into the lung and into the systemic circulation in a lung injury model. Lung injury was induced in 40 anesthetized paralyzed Sprague-Dawley rats (350 +/- 2 g) by hydrochloric acid instillation (pH 1.5, 2.5 ml/kg). Rats were then randomized into five groups (n = 8): (1) high-volume zero PEEP (HVZP): VT, 16 ml/ kg; (2) high-volume PEEP (HVP): VT, 16 ml/kg, PEEP, 5 cm H2O; (3) low-volume zero PEEP (LVZP): VT, 9 ml/kg; (4) low-volume PEEP (LVP): VT, 9 ml/kg, PEEP, 5 cm H2O; (5) same settings as (4) plus a recruitment maneuver performed every hour (LVPR). Respiratory rate was adjusted to maintain normocapnia and fraction of inspired oxygen (FIO2) was 1. Cytokine concentrations (tumor necrosis factor-alpha [TNF-alpha] and macrophage inflammatory protein-2 [MIP-2]) were measured by ELISA. All animals in the LVZP group died before the end of the experiment. After 4 h of ventilation, the HVZP group had similar lung fluid TNF-alpha concentrations compared with the HVP group: 1,861 +/- 333 pg/ml versus 1,259 +/- 189 pg/ml; and much higher serum concentrations: 692 +/- 74 pg/ml versus 102 +/- 31 pg/ml (p < 0.05). An identical pattern was found for MIP 2. These results suggest that the particular ventilatory strategy can affect the release of cytokines into the systemic circulation, a finding that may have relevance for the development of multisystem organ failure. PMID- 10390388 TI - Healthy women's PEF variations with ambient summer concentrations of PM10, PM2.5, SO42-, H+, and O3. AB - The relationship between ambient air pollution and daily change in peak expiratory flow (PEF) was studied in a sample of 473 nonsmoking women (age 19 to 43 yr) in Virginia over summers 1995- 1996. Daily 24-h averages of particulate matter (PM2.5 and PM10), fine particulate sulfate (SO42-) and strong acid (H+), hourly ozone (O3), and select meteorologic variables (e.g., temperature) were collected at a regional outdoor monitoring site. Subjects took PEF measurements twice daily for a 2-wk period using a standard MiniWright peak flow meter. Concurrent measures for summer periods of 24-h PM2.5 (micrograms/m3) ranged from 3.5 to 59.7; H+ (nmol/m3) from 0 to 250; maximal daily 8-h average O3 (ppb) from 17.0 to 87.6. Morning PEF decrements were significantly associated with H+ and PM2. 5. An increase of 50 etamol/m3 of H+ and 10 micrograms/m3 of PM2.5 related to decreases of 0.89 (95% CI = 0.21 to 1.57) and 0.73 (95% CI = 0.07 to 1.38) L/min in morning PEF, respectively. Ozone was the only exposure related to evening PEF with 5-d cumulative lag exposure showing the greatest effect; 7.65 L/ min (95% CI = 2.25 to 13.0) decrease per 30 ppb O3 increase. Separate physiologic effects were observed for summer ambient concentrations of two different pollutants (PEF decrements related to PM2.5 in morning and O3 in evening) at concentrations below the new U.S. EPA 24-h ambient air quality standard for PM2.5 and 8-h standard for O3. PMID- 10390389 TI - The effect of inhaled fluticasone propionate in the treatment of young asthmatic children: a dose comparison study. AB - The response in asthmatic young children to inhaled steroids within the usual pediatric dose range is unknown. We therefore evaluated the dose-related response in young children with moderate asthma to inhaled fluticasone propionate (FP) (delivered via the Babyhaler spacer device) within the pediatric dose range. A total of 237 children (mean age 28 mo, range 12 to 47) with moderate asthmatic symptoms were studied in this multicenter, randomized, double-blind, parallel group, placebo-controlled study of 12 wk treatment following a 4-wk run-in period. The median use of rescue medication was 1 dose in 2 d during the run-in period. FP 50 micrograms twice daily (FP100) and 100 micrograms twice daily (FP200) was compared with placebo inhaled from a pressurized metered-dose inhaler (pMDI) and the Babyhaler spacer device. With FP200 there was a statistically significant improvement from baseline, as compared with the placebo group, in 8 of 10 diary card parameters, including the three symptom domains of wheeze, cough, and breathlessness, and use of rescue medication. FP100 produced a significant reduction in 5 of these 10 parameters, whereas no significant differences were found between the FP200 and FP100. The numbers of patients with at least one exacerbation during treatment with placebo, FP100, and FP200 were 37%, 26%, and 20%, respectively. This difference between placebo and FP200, as well as the dose-related order was significant (p < 0.05). Both FP doses were as well tolerated as placebo over the 12 wk treatment with a similar incidence of adverse effects. Asthmatic symptoms in 1- to 3-yr-old children responded in a significant and dose-related manner to treatment with FP within a pediatric dose range. PMID- 10390390 TI - Increased plasma levels of adrenomedullin in patients with systemic inflammatory response syndrome. AB - We measured the plasma levels of adrenomedullin (AM), a novel vasodilating peptide, in 89 patients with various forms of systemic inflammatory response syndrome (SIRS) and 13 healthy volunteers serving as controls. Plasma levels of AM in SIRS (burns: 20.5 +/- 3. 2 fmol/ml [mean +/- SEM]; pancreatitis: 13.8 +/- 3.8 fmol/ml; trauma: 14.9 +/- 2.5 fmol/ml; traumatic shock: 41.1 +/- 7.8 fmol/ml; severe sepsis: 59.9 +/- 11.2 fmol/ml; septic shock: 193.5 +/- 30.1 fmol/ml) were significantly increased over those of controls (5.1 +/- 0.2 fmol/ml). The patients with traumatic shock or septic shock especially had higher levels of plasma AM than those with trauma or severe sepsis, respectively. These data showed that in patients with SIRS, plasma AM levels increased in proportion to the severity of illness. Subsequently, we measured the plasma levels of mediators such as tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, IL-8, plasminogen activator inhibitor (PAI)-1, and thrombomodulin (TM) in patients with traumatic shock and septic shock. A significant correlation was observed between plasma AM and TNF-alpha levels in patients with septic shock, suggesting an important role for AM as well as of TNF-alpha in the pathophysiology of inflammation. Plasma AM and IL-8 levels correlated positively with Acute Physiology and Chronic Health Evaluation (APACHE) II score, peak multiple organ failure (MOF) score during the first month and prognosis in patients with septic shock, as did plasma IL-6 levels in patients with traumatic shock. The plasma AM level might serve as a useful marker for evaluating the severity of disease and as an early predictor of subsequent organ failure and outcome in septic shock. PMID- 10390391 TI - Increased intrapulmonary oxygen consumption in mechanically ventilated patients with pneumonia. AB - Pulmonary oxygen consumption (V(O2)pulm) is believed to be increased in patients with lung infection. In the present study, VO2pulm was estimated from the difference between total oxygen consumption measured with indirect calorimetry (V(O2)cal) and oxygen consumption assessed with the reverse Fick method (V(O2)Fick). Seventy-five patients requiring mechanical ventilation were included, and were divided for analysis into two groups according to the existence (n = 41) or absence (n = 34) of pneumonia. V(O2)pulm was correlated with various parameters of impaired lung function. To assess the metabolic function of the lung, the differences in lactate and glucose concentrations at different arterial-mixed venous concentrations were determined and transpulmonary lactate flux as well as glucose flux was calculated. As compared with V(O2)pulm in patients without pneumonia (19.4 +/- 1.2 ml/ min/m2), V(O2)pulm was significantly increased in patients with pneumonia (50.7 +/- 1.7 ml/min/m2 (p < 0.001). For intrapatient measurements of V(O2)pulm, a sufficient reproducibility was achieved. V(O2)pulm increased with the lung injury score, number of afflicted lobes, venous admixture, the transpulmonary lactate flux, and the transpulmonary glucose flux, respectively. We speculate that the increased V(O2)pulm of infected lungs is due to different mechanisms, including increased oxidative metabolism by essentially extrapulmonary structures such as neutrophils and macrophages, as well as by changes in the metabolic function of lung tissue itself. PMID- 10390392 TI - Effect of aerosolized uridine-5'-triphosphate on airway clearance with cough in patients with primary ciliary dyskinesia. AB - Primary ciliary dyskinesia (PCD) is a genetic disease characterized by abnormal ciliary structure and function and impaired mucociliary clearance. Because patients with PCD use cough clearance as an airway defense mechanism, we tested the hypothesis that aerosolized uridine-5'-triphosphate (UTP) would improve clearance during cough by its actions to stimulate Cl- secretion and mucin release by goblet cells. We measured clearance during cough in 12 patients with PCD (ages 14 to 71 yr, FEV1 43% to 89% predicted) in a double blind, randomized, crossover study after aerosolization of a single dose of UTP (5 mg/ml, 3.5 ml) or vehicle (0.12% saline, 3.5 ml). Clearance during cough (whole lung) was quantified during and after a series of controlled coughs by measuring the clearance of [99mTc]Fe2O3 particles via gamma camera scanning over 120 min. Safety parameters were recorded during and after drug delivery. Aerosolized UTP improved whole-lung clearance during cough as compared with vehicle (from 0 to 60 min: 0.40 +/- 0.07%/min [UTP] versus 0.26 +/- 0. 04%/min [vehicle] [mean +/- SEM], p = 0.01), and from 0 to 120 min: 0.38 +/- 0.05%/min [UTP] versus 0.25 +/- 0.04%/ min [vehicle], p = 0. 02). Aerosolized UTP is safe, with no serious adverse effects. Whole-lung clearance during cough in patients with defective ciliary function is enhanced after inhalation of UTP. PMID- 10390393 TI - Risk factors from childhood to adulthood for bronchial responsiveness at age 32 42 yr. AB - Bronchial responsiveness (BR) is an important risk factor for the development and outcome of asthma. This study assessed childhood risk factors for both the severity of BR in adulthood and either improvement or worsening of BR over time. Finally, we studied cross-sectional risk factors of BR in adulthood. Between 1966 and 1969, 119 allergic asthmatic children (5-14 yr of age) were studied. Of these, 101 (85%) subjects were reinvestigated at age 22-32 yr (visit 2), and at age 32-42 yr (visit 3). Spirometry, PC10 histamine, skin tests, blood eosinophils, and serum total IgE were measured and a questionnaire was used. Higher FEV1 values in childhood were associated with less severe BR at age 32-42 yr independent of other potential risk factors. Larger increases in FEV1 values both from visit 1 to 2 and from visit 2 to 3, a longer time interval from visit 1 to 3, and having pets in childhood were associated with less severe BR at age 32 42 yr. The same factors were found to be associated with less deterioration of BR from visit 2 to 3. In nonsmokers a higher IgE level at visit 2 was a risk factor for an increase in BR. At age 32-42 yr, a low level of lung function and the presence of asthma symptoms were associated with more severe BR, and older age and having pets were associated with less severe BR. IgE was related to more severe BR only in nonsmokers. CONCLUSIONS: A lower lung function in childhood and less improvement in FEV1 over time were associated with more severe BR in adulthood. PMID- 10390394 TI - Systemic effects of inhaled fluticasone propionate and budesonide in adult patients with asthma. AB - We assessed the systemic effects of budesonide (BUD) and fluticasone propionate (FP) in 23 patients with asthma, using a double-blind, placebo-controlled, double dummy, and cross-over design. The following five treatments were given in a randomized order for 1 wk with a washout period in between of 2 wk: (1) placebo; (2) FP, 200 micrograms twice a day, inhaled from a Diskhaler; (3) FP, 1,000 micrograms twice a day, inhaled from a Diskhaler; (4) BUD, 200 micrograms twice a day, inhaled from a Turbuhaler; and (5) BUD, 800 micrograms twice a day, inhaled from a Turbuhaler. The primary variable was the area under the curve of serum cortisol versus time (AUC0-20), derived from serum samples taken every 2 h over a 20-h period following the last evening dose at 10:00 P.M. The lower doses of BUD and FLU did not cause any adrenal suppression. Compared with placebo, however, FP (1, 000 micrograms, twice daily and BUD (800 micrograms, twice daily) decreased the AUC0-20 by 34 and 16%, respectively. Fluticasone (1,000 micrograms, twice daily) was more suppressive than BUD (800 micrograms, twice daily) (p = 0.0006). The FEV1, measured the morning after the last inhalation, was significantly higher after the active treatments, compared with placebo (p < 0.02), but did not differ between all active treatments. We conclude that high doses of BUD and FP (in particular the latter), inhaled via their respective dry powder inhalers for 1 wk, result in a measurable systemic activity in patients with asthma. PMID- 10390395 TI - A comparison of pulmonary arterial occlusion algorithms for estimation of pulmonary capillary pressure. AB - Using the arterial occlusion method, we compared five literature-based estimates of pulmonary capillary pressure (Ppc) with the corresponding double occlusion pressures (Pdo) in anesthetized dogs whose chests had been closed after sternotomy for instrumentation. Arterial occlusions were performed with a balloon tipped pulmonary artery catheter that housed pressure transducers immediately proximal and distal to the balloon. Separation of the proximal and distal pressure waveforms during balloon inflation allowed us to precisely define the moment of occlusion. We fit a monoexponential curve to pressure data beginning 200 ms after the onset of occlusion and a biexponential curve to data beginning at the instant of occlusion, with data obtained over a range of vascular states (control, serotonin infusion, histamine infusion). In addition, we investigated the use of sampling of the raw data to estimate capillary pressure. Three of the five literature-based estimates of Ppc yielded values similar to Pdo. The optimal (least average difference from Pdo) interpolation/extrapolation time points of the curve fits varied, depending on the type of curve fitting and the state of the pulmonary vasculature. We also determined that a close approximation of Pdo may be derived from the raw data, as an alternative to exponential curve fitting. PMID- 10390396 TI - Progressive mechanical ventilatory constraints with aging. AB - To investigate the progressive nature of mechanical ventilatory constraints with aging, we studied 20 young (age 39 +/- 3 yr), 14 senior (70 +/- 2 yr), and 11 elderly (88 +/- 2 yr) men and women during exercise. All subjects had normal pulmonary function and performed graded cycle ergometry to exhaustion. Minute ventilation (V E), lung volume, and expiratory airflow limitation (EAFL) were measured during each 1-min increment in work rate. Data were analyzed by two-way analysis of variance (ANOVA; age x gender) at rest, ventilatory threshold (VTh), and peak exercise. If an interaction was present, each gender was analyzed with a one-way ANOVA. Aging resulted in an increased V E for a given submaximal work rate, although V E during peak exercise was lowest in the elderly group (p < 0.01). End-expiratory lung volume (EELV, % of TLC) in men increased progressively with age and all groups were different at VTh (p < 0.01) and peak exercise (p < 0.01). In women, EELV (% of TLC) also increased with aging, the senior and elderly subjects had a greater EELV at VTh (p < 0.01) and peak exercise (p < 0.01) than the young group. Additionally, the normal decrease in EELV during the early stages of exercise was not observed in elderly subjects. End-inspiratory lung volume (EILV) also progressively increased with aging; senior and elderly subjects had a higher EILV at rest (p < 0.05), VTh (p < 0.01), and peak exercise (p < 0.01) than young subjects. EAFL (% of VT) increased with aging; elderly subjects experienced greater EAFL at rest (p < 0.05), VTh (p < 0.01), and peak exercise (p < 0.01) than both young and senior subjects. We conclude that mechanical ventilatory constraints are progressive with aging, elderly subjects demonstrating marked mechanical ventilatory constraints during exercise. The impact of these constraints on exercise tolerance cannot be determined from this investigation and remains unclear. PMID- 10390397 TI - Tuberculosis prevention in methadone maintenance clinics. Effectiveness and cost effectiveness. AB - To determine the effectiveness and cost-effectiveness of a program to provide screening for tuberculosis infection and directly observed preventive therapy (DOPT) in methadone maintenance clinics, we determined completion rates of screening for tuberculosis infection, medical evaluation, and preventive therapy, as well as the number of active tuberculosis cases and tuberculosis-related deaths prevented, in five clinics in San Francisco, California. Between 1990 and 1995, a total of 2,689 clients (of whom 18% were HIV-seropositive) were screened at least once. Of eligible clients, 99% received tuberculin skin tests, 96% received a medical examination, 91% began isoniazid preventive therapy, and 82% completed preventive therapy. Program effectiveness was enhanced by close collaboration between public health and methadone maintenance programs and the use of incentives and enablers. Over a 3-yr follow-up period, only one verified case of tuberculosis was reported among clients with a positive tuberculin skin test, thereby preventing as much as 95% of expected tuberculosis cases. Over 10 yr, we estimate the program would prevent 30.0 (52%) of 57.7 expected cases of tuberculosis, and 7.6 (57%) of 13.4 expected tuberculosis-related deaths. The program cost $771,569, but averted an estimated $876,229, for a net savings of $104,660 (average of $3, 724 per case prevented). Our study demonstrates that when effectively implemented, screening for tuberculosis infection and DOPT in methadone maintenance clinics is a highly cost-effective approach to prevent tuberculosis. PMID- 10390398 TI - Quantitation of inflammatory responses to bacteria in young cystic fibrosis and control patients. AB - Recent studies suggest that inflammation plays a role in the pathogenesis of lung disease in cystic fibrosis (CF). The goal of the present study was to quantitatively compare bronchoalveolar lavage fluid (BALF) inflammation and its relation to bacterial infection, between children with CF and children with other chronic respiratory problems. Differential cell counts, immunoreactive interleukin 8 (IL-8), and quantitative bacterial cultures were done in BALF from 54 CF (median age 1.8 yr) and 55 control patients (median age 1.0 yr) who underwent bronchoscopy for clinical indications. Among infected CF patients, those with Pseudomonas aeruginosa did not have more inflammation than those without P. aeruginosa. The ratio of neutrophils or of IL-8 to bacteria in BALF was significantly greater for CF patients compared with control subjects, regardless of pathogen. Calculation of linear regression for either neutrophils or IL-8, as a function of bacterial quantity, yielded positive slopes for both CF and control patients, but with significant elevations for CF. We conclude that the inflammatory response to bacterial infection is increased or prolonged in CF compared with control patients, and that this increase is not necessarily due to pathogens specific for CF (e.g., P. aeruginosa). These data may provide further rationale for anti-inflammatory therapy early in CF. PMID- 10390399 TI - Randomized trial of prolonged chloroquine therapy in advanced pulmonary sarcoidosis. AB - Sarcoidosis may cause severe ventilatory impairment requiring corticosteroid treatment. Chloroquine (CQ) can be an effective treatment for lung sarcoidosis with few side effects, but has not been accepted as standard therapy. We investigated the benefits of prolonged CQ therapy in 23 symptomatic patients with biopsy-proven pulmonary sarcoidosis (duration, >/= 2 yr). Patients were initially treated for 6 mo with CQ, 750 mg/d, tapering every 2 mo to 250 mg/d. Eighteen patients were then randomized to either a Maintenance group (CQ, 250 mg/d) or to an Observation group (no CQ). After the initial treatment, significant improvement was observed in symptoms, pulmonary function, angiotensin-converting enzyme, and lung gallium scan. Patients randomized to the Maintenance group showed a slower decline in pulmonary function (FEV1, 51.4 +/- 28.2 ml/yr [Maintenance] versus 196.3 +/- 33.4 ml/yr [Observation], p < 0.02) and had fewer relapses: 2 of 10 patients in the Maintenance group at 29.5 +/- 4.9 mo versus 6 of 8 patients in the Observation group at 15.5 +/- 2.9 mo. Adverse effects were seen mainly during high-CQ dosage. We conclude that CQ should be an important consideration for the treatment and maintenance of chronic pulmonary sarcoidosis. PMID- 10390400 TI - Nocturnal oxygenation and prognosis in Duchenne muscular dystrophy. AB - REM-related oxygen desaturation occurs in advanced Duchenne muscular dystrophy (DMD) and might be an independent predictor of disease progression. We have followed 18 patients for 10 yr after an initial respiratory sleep study or until death or onset of nasal ventilation. We measured baseline spirometry, blood gas tensions, maximal respiratory pressures, and body mass index. In 11 cases, VC was recorded serially. Median survival was 50 (range, 13 to 89) mo from initial study and unrelated to age at time of study, BMI, or mouth pressures but correlated with PaCO2 (r = -0.72, p < 0.005, n = 17), minimal nocturnal SaO2 (r = 0.62, p < 0.007, n = 18) and VC (r = 0. 65, p < 0.005, n = 17). Cox regression analysis showed a VC of less than 1 L at the time of study to be the best single predictor of subsequent survival. The only measure associated with age of death was the age at which the VC fell below 1 L (r = 0.79, p < 0.004). These data suggest measurement of PaCO2 or serial assessment of VC should be studied further as valid methods of assessing prognosis in DMD. PMID- 10390401 TI - Measurement of sputum Mycobacterium tuberculosis messenger RNA as a surrogate for response to chemotherapy. AB - Effective treatment regimens for pulmonary tuberculosis are difficult to assess because of the slow growth rate of Mycobacterium tuberculosis in culture and its protracted clearance from sputum. A rapid method that reflects effective antimicrobial activity would markedly advance evaluation of treatment and promote the assessment of new antituberculosis drugs. Conventional methods measure the progressive reduction of numbers of acid-fast bacilli in the sputum smear and the clearance of organisms in sputum culture. In this study, we measured levels of M. tuberculosis 85B (alpha antigen) messenger RNA (mRNA), 16S ribosomal RNA (rRNA), and IS6110 DNA in patients' sputa to ascertain whether they could serve as potential surrogate markers of response to chemotherapy. Sputum specimens were sequentially collected for up to a year from 19 smear-positive pulmonary tuberculosis patients receiving an optimal drug treatment regimen. Nucleic acids were isolated from these specimens, and two M. tuberculosis molecular targets (mRNA, DNA) were quantified, using the ABI Prism 7700 Sequence Detection System. The Mycobacterium genus-specific 16S rRNA was quantified with a limiting dilution RT-PCR assay. Results show that levels of 85B mRNA declined after initiation of therapy, as did viable M. tuberculosis colony counts, with 90% of patients becoming negative for both markers after 2 mo of treatment. The rapid disappearance of M. tuberculosis mRNA from sputum suggests that it is a good indicator of microbial viability and a useful marker for rapid assessment of response to chemotherapy. PMID- 10390402 TI - The cost of asthma in the emergency department and hospital. AB - Treatment of asthma in the emergency department (ED) or hospital accounts for a significant portion of total treatment costs; however, little is known about the specific resources consumed. The purpose of this study was to estimate the type and amount of resources consumed for an asthma event requiring ED visit and/or hospitalization. Between October 1, 1996 and September 30, 1997, occurrences of asthma as a primary diagnosis were identified at 27 hospitals' emergency departments within Premier's Perspective Comparative Database. Patients visiting the ED could either be treated and released or admitted to the hospital. A total of 3,223 patients (age >/= 18 yr) were identified, with 1,074 (33.3%) requiring hospitalization. For the 2,149 patients who visited the ED only, the average visit cost was $234.48. For hospitalized patients, the average length of stay was 3.8 d, and the cost was $3,102.53. Nursing care was the source of the majority of hospital costs for asthma (43.6%), respiratory therapy (13.6%), and medications (10.4%). For adult asthma patients requiring hospitalization, the total cost is high and resources consumed are unavoidable. Thus, a continuum of care aimed at appropriate asthma management, especially in the elderly, could result in substantial cost savings over those aimed at reducing inpatient utilization of care. PMID- 10390403 TI - Increased 8-isoprostane, a marker of oxidative stress, in exhaled condensate of asthma patients. AB - Oxidative stress has an important role in the pathogenesis of asthma. 8 Isoprostane is a prostaglandin (PG)-F2-like compound belonging to the F2 isoprostane class that is produced in vivo by the free radical-catalyzed peroxidation of arachidonic acid. 8-Isoprostane is a biomarker of oxidative stress, and its concentration is increased in the bronchoalveolar lavage fluid of patients with interstitial lung diseases. We measured 8-isoprostane concentrations in exhaled breath condensate in healthy subjects and in patients with mild (steroid naive, n = 12), moderate (inhaled steroid treatment, n = 17), and severe asthma (oral steroid treatment, n = 15). We also measured exhaled carbon monoxide (CO) and nitric oxide (NO), which may also reflect oxidative stress in the airways. 8-Isoprostane was detectable in breath condensate of normal subjects (15.8 +/- 1.6 pg/ml), and was increased in the breath condensate of patients with mild (33.7 +/- 2.8, p < 0.001), moderate (38.3 +/- 3.7 pg/ml, p < 0. 001), and severe asthma (48.9 +/- 5.0 pg/ml, p < 0.001). There was a positive correlation (r = 0.68, p < 0.05) of 8-isoprostane with NO, but not with CO, in the exhaled air of patients with mild asthma, but not in that of patients with moderate or severe asthma. There was no correlation between 8-isoprostane and lung function tests in any group of patients. Our study shows that oxidative stress is increased in asthmatic subjects as reflected by 8-isoprostane concentrations in breath condensate. PMID- 10390404 TI - Thermally induced asthma and airway drying. AB - The purpose of this study was to determine whether mucosal dehydration causes thermally induced asthma. To provide data on this point, we studied the effects on lung function of progressive water loss (WL) from the respiratory tract by having eight subjects perform isocapnic hyperventilation for 1, 2, 4, and 8 min at a constant level (V E = 57.5 +/- 6.3 L/min [mean +/- SEM]) while they breathed dry air at frigid (TI = -12.5 +/- 2.7 degrees C) (cold trial) and ambient (24.3 +/- 0.7 degrees C) (warm trial) temperatures. Expired temperatures (TE) were continuously monitored, and WL from the intrathoracic airways was calculated from published relationships. FEV1 was measured before and after each challenge. Each inspirate produced stimulus-response decrements in FEV1, but the effect of cold air was greater (% Delta cold8min = 30.0 +/- 4.7%, warm = 16.0 +/- 4.4%; p = 0.01). Water loss, however, was significantly less in the cold experiment because TE was lower (WL cold8min = 4.8 +/- 0.4 g, warm = 7.1 +/- 0.7 g; p = 0.001; TE cold8min = 22.8 +/- 2.3 degrees C, warm 30.9 +/- 1.5 degrees C; p = 0.003). The FEV1 decreased as WL rose, but the largest intrathoracic losses were associated with the smallest obstructive response (% DeltaFEV1 cold8min = 30%, WL = 4.7 mg; % DeltaFEV1 warm8min = 16%, WL = 7.1 mg; p = 0.002). These data show that removal of water from the lower respiratory tract, and by inference the development of a hyperosmolar periciliary fluid, do not appear to be the primary causes of thermally induced asthma. PMID- 10390405 TI - Predictors of repeated wheeze in the first year of life: the relative roles of cockroach, birth weight, acute lower respiratory illness, and maternal smoking. AB - While more than 80% of childhood asthmatics are allergic to one or more inhaled allergens, the role of inhaled allergens in the induction of wheeze in the first year of life is unknown. In a prospective birth-cohort study of 499 children of asthmatic/allergic parents from metropolitan Boston, we examined home allergen concentrations measured within the first 3 mo of life as predictors of repeated wheeze episodes in the first year of life. In multivariate analyses adjusting for maternal asthma and dog in the home, predictors of two or more wheeze episodes in the first year of life included maternal smoking during pregnancy (relative risk [RR] = 1.83; 95% confidence limit [CL]: 1.12, 3.00), lower respiratory illness in the first year of life (croup, bronchitis, bronchiolitis, or pneumonia) (RR = 2.25; 95% CL:1.58, 3.19), low birthweight (RR = 1.28, 95% confidence interval [CI]: 1.04, 1.58 for an interquartile difference), and Bla g 1 or 2 (cockroach) allergen level in the family room > 0.05 U/g dust (RR = 1.76; 95% CL: 1.20, 2.57). Cockroach allergen in the family room and repeated wheeze remained significant after adjustment for socioeconomic factors including race and income (RR = 1.63; 95% CL: 1.05, 2.55). It is unknown whether the association between cockroach and repeated wheeze in infancy represents a cockroach-related increased risk of bronchial inflammation through nonallergenic or allergenic mechanisms. PMID- 10390406 TI - A dose-dependent effect of the novel inhaled corticosteroid ciclesonide on airway responsiveness to adenosine-5'-monophosphate in asthmatic patients. AB - Inhaled corticosteroids decrease airway responsiveness in asthma partly through suppression of airway inflammation. We have previously demonstrated that inhaled budesonide reduced airway responsiveness to the mast cell stimulus adenosine-5' monophosphate (AMP) to a threefold greater extent than to methacholine and sodium metabisulfite, suggesting that AMP responsiveness may be a more sensitive marker of airway inflammation and steroid action in order to assess a dose-response relationship. To investigate this, we studied the effects of three doses of the novel corticosteroid ciclesonide (50 micrograms, 200 micrograms, and 800 micrograms) inhaled as a dry powder twice daily on airway responsiveness to AMP and inflammatory parameters in induced sputum. In a three-parallel-dose group, double-blind, placebo-controlled, randomized, crossover study, with a washout period of 3 to 8 wk, a total of 29 patients with mild to moderate allergic asthma underwent AMP challenge and sputum induction before and after 14 d of treatment with ciclesonide or matched placebo. Compared with placebo, ciclesonide 100 micrograms, 400 micrograms, and 1,600 micrograms daily reduced airway responsiveness to AMP by 1.6 (95% confidence interval [CI], -0.1 to 3.4, not significant [NS]), 2.0 (95% CI, 0.4 to 3.6, p < 0.05), and 3.4 (95% CI, 2.3 to 4. 4, p < 0.05) doubling doses, respectively, and this reduction in airway responsiveness was dose-dependent (p = 0.039). A significant reduction in the percentage of eosinophils in induced sputum was observed after 400 micrograms and 1,600 micrograms daily ciclesonide (p < 0. 05), but this was not dose-dependent. Sputum eosinophil cationic protein (ECP) was significantly reduced after 400 micrograms daily ciclesonide only (p < 0.05). Thus, in patients with mild to moderate asthma, assessment of airway responsiveness to AMP, rather than inflammatory parameters in induced sputum, represents a sensitive method to evaluate a dose-response relationship of an inhaled corticosteroid and may have applications in evaluating other novel inhaled corticosteroids. PMID- 10390407 TI - Comparison of the relative efficacy of formoterol and salmeterol in asthmatic patients. AB - Studies performed on airway smooth muscle in vitro have indicated that salmeterol is a partial agonist on the beta2-receptor in comparison to formoterol. In the present study we evaluated whether these pharmacological differences between salmeterol and formoterol also are applicable to asthmatic patients. The protective effects by increasing cumulative doses of formoterol (12, 60, 120 micrograms) and salmeterol (50, 250, 500 micrograms) on methacholine-induced bronchoconstriction were evaluated in a double-blind, crossover, placebo controlled design. Patients were regularly treated with salbutamol 200 micrograms twice daily during the study period, to avoid variability in beta2-adrenoceptor tolerance. S-potassium, heart rate corrected Q-T interval (Q-Tc), and tremor score were followed as measures of systemic effects. Formoterol dose-dependently protected against methacholine responsiveness (4.6 doubling doses after 120 micrograms). Salmeterol, however, showed a flatter dose-response curve, and a significantly weaker maximal protective effect (2.8 doubling doses after 250 micrograms). Formoterol caused a significantly higher tremor score and a larger drop in S-potassium than salmeterol at the highest doses. These data show that salmeterol is a partial agonist on the beta2-receptor in relation to formoterol in human airways in vivo. Further studies are required to document the clinical consequences of this finding, for example in severe asthmatic patients. PMID- 10390408 TI - Regional expansion of oleic acid-injured lungs. AB - It has been suggested that dependent regions of an injured lung are collapsed and subject to cyclic reopening and collapse during mechanical ventilation. To test this hypothesis, we measured both temporal and spatial heterogeneity of lobar expansion in oleic acid (OA)-injured dogs. Regional volumes were measured in nine dogs (seven supine and two prone) during closed loop sinusoidal oscillations of the lungs before and after OA injury using the parenchymal marker technique. In contrast to computer tomography, the parenchymal marker technique provides absolute measures of regional tissue dimensions as opposed to relative measures of regional air to liquid content. The experiments generated three major findings: (1) OA injury did not lead to the collapse of dependent lung units at FRC, (2) OA injury did not steepen the vertical gradient in regional lung volumes at FRC, and (3) during sinusoidal oscillation of the OA-injured lungs from FRC, dependent regions did not undergo cyclic reopening and collapse. On the basis of these results, we propose an alternative mechanism for the topographic variability in regional impedances and lung expansion after injury, namely liquid and foam in conducting airways. PMID- 10390409 TI - Smoking and airway hyperresponsiveness especially in the presence of blood eosinophilia increase the risk to develop respiratory symptoms: a 25-year follow up study in the general adult population. AB - Airway hyperresponsiveness (AHR) constitutes a risk for development of respiratory symptoms. We assessed whether blood eosinophilia (>/= 275 eosinophils/microliters), skin test positivity (sum score >/= 3) and cigarette smoking (never, ex-smoker, 1-14 cig/d, 15-24 cig/d, >/= 25 cig/d) at the first of two successive surveys are related to the development of respiratory symptoms (chronic cough or phlegm, bronchitis, persistent wheeze, dyspnea, and asthma) at the second survey, and whether these relations are the same in subjects with (PC10 55 yr (n = 4). Therapeutic manipulation resulted in improvement in the PaO2/FIO2 ratio in 20 donors (Group 3) who would not otherwise have been used. Immediate and 24 h postoperative gas exchange and length of intensive care unit (ICU) stay was not different for recipients from donors from all three groups. Overall survival was 94% at 30 d, 83% at 1 yr, 70% at 2 yr, and 62% at 3 yr and was not significantly different from the three groups. We conclude that organ utilization can be maximized by therapeutic manipulation and utilization of marginal donors without compromising results from transplantation. PMID- 10390411 TI - The pathophysiology of pulmonary diffusion impairment in human immunodeficiency virus infection. AB - Numerous reports have demonstrated that prior to the development of acquired immunodeficiency syndrome (AIDS)-related pulmonary complications, human immunodeficiency virus-positive (HIV+) individuals commonly develop unexplained reductions in pulmonary diffusing capacity (DLCO). The potential relevance of this observation is underscored by recent data demonstrating that reductions in DLCO independently predict the subsequent development of opportunistic pneumonia. To delineate the alterations in gas exchange associated with HIV, we investigated a group of HIV+ subjects with unexplained reductions in DLCO, using high resolution computed tomography (HRCT) of the chest and a separation of diffusing capacity into its membrane (Dm) and capillary blood volume (Vc) components. We compared this abnormal group with HIV+ subjects with more normal gas exchange and also with a group of HIV- volunteers matched for age and smoking history. Compared with other groups, the HIV+ group with diffusion impairment demonstrated prominent reductions in Vc, despite a well-preserved total lung capacity (TLC). HRCT demonstrated virtually no evidence of interstitial fibrosis in any HIV+ subject, but evidence of early emphysema that significantly correlated with DLCO. Our results suggest that the previously reported impairment in pulmonary gas exchange in the HIV+ population involves loss of Vc and likely represents the development of early emphysema. PMID- 10390412 TI - Prevalence of tumor necrosis factor-alpha and angiotensin converting enzyme polymorphisms in mild/moderate and fatal/near-fatal asthma. AB - Allele 2 of the polymorphism at position -308 in the promoter of the tumor necrosis factor alpha (TNF-alpha) gene, and the D allele of the angiotensin converting enzyme (ACE) gene, have been associated with asthma. We hypothesized that genotypes containing these alleles would show an increased prevalence in asthmatic as compared with nonasthmatic individuals, and would be associated with asthma severity. Polymerase chain reaction-based assays were developed to determine TNF-alpha and ACE genotypes among subjects with mild/moderate asthma (n = 92), fatal/near-fatal asthma (n = 159), no asthma (n = 43), and random population controls (n = 252). The TNF-alpha -308 polymorphism was increased in both subjects with mild/moderate (p = 0.03) and those with fatal/near fatal asthma (p = 0.02) versus those without asthma, and in all subjects with asthma versus random population controls (p = 0.02). The mild/moderate group was subdivided into subjects with mild (n = 43) and those with moderate (n = 33) asthma. TNF-alpha -308 was increased in the moderately asthmatic versus the nonasthmatic subjects (p = 0.003), and in the mildly asthmatic subjects (p = 0.01). However, TNF-alpha -308 was not significantly more prevalent in the fatal/near-fatal than in the mild/moderate asthmatic group. The ACE-D allele did not show an association with either asthma or asthma severity. We conclude that the TNF-alpha -308 polymorphism may be a risk factor for asthma but does not increase the risk of a fatal or a near-fatal asthma attack, whereas the ACE polymorphism is not associated with asthma in this population. PMID- 10390413 TI - Antibacterial components in bronchoalveolar lavage fluid from healthy individuals and sarcoidosis patients. AB - Antibacterial peptides and proteins are an integral part of the epithelial defense barrier that provides immediate protection against bacterial invasion. In humans, alpha-defensins are mainly bactericidal effectors in circulating granulocytes, beta-defensin-1 is synthesized in epithelial cells, and LL-37 is produced in granulocytes but is also induced in skin epithelia during inflammation. To investigate the importance of these defense effectors in disease, we analyzed bronchoalveolar lavage fluid (BALF) for bactericidal activity. Antibacterial activity was found in BALF material from healthy individuals and sarcoidosis patients, with enhanced activity in BALF from the patients. The activity was present as several antibacterial components, of which we have so far characterized LL-37, lysozyme, alpha-defensins, and antileukoprotease. In addition, the antibacterial peptide LL-37 was located in alveolar macrophages, bronchial epithelial cells, and bronchial glands, suggesting that it has a defensive role in airway mucosa. In conclusion, the airway epithelium is protected by a complex antibacterial defense system. This is activated in sarcoidosis, and may explain why these patients seldom develop severe respiratory tract infections. PMID- 10390414 TI - Cyclooxygenase-2 mRNA is downexpressed in nasal polyps from aspirin-sensitive asthmatics. AB - Exogenous prostaglandin E2 (PGE2) given by inhalation almost completely abrogates aspirin-induced asthma and the accompanying increase in cysteinyl-leukotrienes production. Cyclooxygenase (COX) may be present in cells in both constitutive (COX-1) and inducible (COX-2) forms. To increase the production of the potentially protective endogenous PGE2, COX-2 should be upregulated. We hypothesize that an abnormal regulation of COX-2 will predispose patients with asthma to develop aspirin-intolerant asthma/rhinitis (AIAR). We therefore examined the expression of COX-2 messenger RNA (mRNA) in healthy nasal mucosa (n = 11) and in nasal polyps from both patients with AIAR (n = 8) and those with aspirin-tolerant asthma/rhinitis (ATAR) (n = 20). After total mRNA extraction, COX-1 and COX-2 mRNA expression were measured using a reverse transcriptase (RT) semiquantitative PCR technique. Hybrid primers of COX-1. glyceraldehyde-3 phosphate dehydrogenase (GAPDH) or COX-2. GAPDH were used to create PCR products that were cloned and used as internal standard controls in the competitive PCR reaction. Results are presented as mean +/- standard error of 10(6) molecules of mRNA/micrograms of total RNA. No differences in COX-1 mRNA expression were found between nasal mucosa and nasal polyps from both patients with ATAR and those with AIAR. However, COX-2 mRNA expression in nasal polyps from the AIAR group (0.38 +/ 0.10) was markedly and significantly lower than in polyps from the ATAR group (2.93 +/- 0. 52, sevenfold, p < 0.0001) and nasal mucosa (2.10 +/- 0.54, sixfold, p < 0.01). These findings suggest that an inadequate COX-2 regulation may be involved in AIAR. PMID- 10390415 TI - Variable airway responsiveness to inhaled lipopolysaccharide. AB - Individuals exposed to inhaled endotoxin (lipopolysaccharide [LPS]) can develop airway symptomatology and exacerbations of asthma. Moreover, among those occupationally exposed to organic dusts, the progression of airflow obstruction is related to the endotoxin concentration in the bioaerosol. Not everyone exposed to high concentrations of LPS develops these problems. To determine whether individuals express a differential response to inhaled LPS, we challenged 72 healthy volunteers with increasing doses of LPS. Airflow was assessed after each dose and the protocol was terminated for decline in FEV1 >/= 20%. Marked differences in the response to inhaled LPS were observed: eight "sensitive" subjects had at least 20% decline in their FEV1 after inhaling 6.5 micrograms or less of LPS, whereas 11 "hyporesponsive" subjects maintained an FEV1 >/= 90% of their baseline even after inhaling 41.5 micrograms of LPS. Serial testing demonstrated that the response to inhaled LPS is reproducible. Sensitive subjects were more commonly female and hyporesponsive subjects were more often male (p = 0.016). Peripheral blood monocytes from hyporesponsive subjects, compared with sensitive subjects, released less interleukin (IL)-6 and IL-8. These findings demonstrate that an LPS phenotype can be reproducibly elicited in humans, which creates an opportunity to identify genes involved in this response to inhaled LPS. PMID- 10390416 TI - Allergen-induced eosinophil cytolysis is a primary mechanism for granule protein release in human upper airways. AB - Cytotoxic eosinophil granule proteins are considered important in the pathogenesis of allergic airway diseases such as rhinitis and asthma. To explore the cellular mechanisms behind eosinophil granule release in human allergic airways, 16 symptom-free patients with seasonal allergic rhinitis were challenged daily with allergen during 1 wk. Nasal lavage samples and biopsies, obtained before and 24 h after the last allergen exposure, were processed for immunohistochemical and electron microscopic analysis. The allergen challenges produced nasal symptoms, marked tissue eosinophilia, and an increase in lavage fluid levels of eosinophil cationic protein (ECP). The nasal mucosa areas with intense extracellular immunoreactivity for ECP were associated with abundant free eosinophil granules. Electron microscopy confirmed the free granules and revealed that all mucosal eosinophils were involved in granule release, either by cytolysis (33%) or piecemeal degranulation (PMD) (67%). Resting or apoptotic eosinophils were not observed. Cytolytic eosinophils had less signs of intracellular granule release (p < 0. 001) and a higher content of intact granules (p < 0.001) compared with viable eosinophils in the same tissue. This study demonstrates eosinophil cytolysis (ECL) as a distinct mechanism for granule mediator release in human allergic airway mucosa. The nature and extent of the ECL and its product (i.e., protein-laden extracellular granules) indicate that allergen-induced cytolysis is a primary and major mechanism for the release of eosinophil proteins in human allergic airway inflammation in vivo. PMID- 10390417 TI - Increased airway smooth muscle in sudden infant death syndrome. AB - The underlying pathophysiological mechanism behind death in the sudden infant death syndrome (SIDS) is uncertain. Although infants dying of SIDS frequently have a postmortem examination performed, no specific diagnostic pathology in any organ system has been identified. Previous theories relating to the cause of death in SIDS have included increased lower airway closure. We examined the airway morphometry of 57 infants who died of SIDS and compared these findings with those obtained from 21 age-matched infants who had died of non-SIDS causes. Airway wall dimensions, epithelial thickness, and the area of smooth muscle within the airway wall were measured. Airways from infants who died of SIDS showed a significantly higher proportion of airway smooth muscle than control airways when corrected for age and sex (p < 0.01). There was no significant difference between the groups for wall thickness or epithelial thickness. Increased airway smooth muscle in infants who have died of SIDS may contribute to excessive airway narrowing, raising the possibility that the cause of death in this condition is related to abnormalities in lower airway function. PMID- 10390418 TI - Novel assessment of acute lung injury by in vivo near-infrared spectroscopy. AB - We investigated the feasibility and validity of near-infrared (NIR) spectroscopy for evaluation of acute lung injury (ALI). In an in vitro model simulating the spectrophotometric characteristics of the lung, NIR spectroscopy could precisely detect changes in water volume, suggesting its ability to assess the extent of pulmonary edema caused by ALI. The different grades of ALI were induced in rats by administering oleic acid and varying the pulmonary ventilation conditions, and NIR spectroscopy was employed to determine lung water content and hemoglobin (Hb) oxygenation of the lungs. NIR spectroscopy detected increased water content even in histologically mild ALI. The changes in lung water content measured by NIR spectroscopy were significantly correlated with gravimetric lung water content (r = 0.877, p < 0.0001). Deoxy-Hb measured by NIR spectroscopy consistently reflected the histological changes in the lungs, and the deoxy-Hb levels correlated with changes in SaO2 (r = -0.798, p < 0.0001). These findings demonstrate that NIR spectroscopy can evaluate lung water content and Hb oxygenation quantitatively, and may be a useful tool for assessing pathological status in ALI. PMID- 10390419 TI - Tissue inhibitor of metalloproteinase-1 levels in bronchoalveolar lavage fluid from asthmatic subjects. AB - Airway remodeling is a well-recognized feature in patients with chronic asthma. The accumulation in the submucosa of fibrous proteins that are substrates of matrix metalloproteinases (MMP), and the demonstration of increased levels of MMP 9 in bronchoalveolar lavage fluid, prompted us to determine whether there was an imbalance between MMPs and tissue inhibitors of metalloproteinase (TIMP) in such patients. We investigated the presence of TIMPs and other MMPs. TIMP levels were compared with those of all MMPs and inflammatory cytokines. Adults with stable asthma, either untreated or treated with glucocorticoids (GCs), were enrolled. Healthy nonsmokers served as a control population. MMPs and TIMPs were identified through zymography or immunoblotting. TIMPs, MMPs, and cytokines were measured with enzyme immunoassays. TIMP-1 levels were significantly higher in untreated asthmatic subjects than in GC-treated subjects or controls (p < 0.0001), and were far greater than those of MMP-1, MMP-2, MMP-3, and MMP-9 combined. TIMP-2 was undetectable. TIMP-1 levels were correlated with levels of interleukin-6 (p < 0.012) and the number of alveolar macrophages recovered (p < 0.005). This observation has important implications, since an excess of TIMP-1 could lead to airway fibrosis, a hallmark of airway remodelling in patients with chronic asthma. PMID- 10390420 TI - Suplatast tosilate inhibits late response and airway inflammation in sensitized guinea pigs. AB - The effect of suplatast tosilate, which has been proven to inhibit T-cell synthesis of IL-4 and IL-5, on the response to antigen inhalation challenge was investigated in sensitized guinea pigs. The animals were given an oral dose of 30 or 100 mg/kg of suplatast or vehicle (distilled water) daily for 1 wk before antigen challenge. Measurement of pulmonary resistance for 6 h was followed by bronchoalveolar lavage and lung fixation. After antigen challenge, all guinea pigs in the vehicle group displayed dual-phase airway obstruction and accumulation of eosinophils and lymphocytes in the airways. After 1 wk of treatment with the high dose of suplatast, the late asthmatic response and the recruitment of eosinophils and lymphocytes into the airways were significantly inhibited, but the early asthmatic response was not affected. In situ hybridization revealed that challenge-induced increases in IL-5 mRNA-positive cells in lung tissue were significantly inhibited after treatment. Thus, suplatast inhibited airway obstruction in the late phase by specifically inhibiting the inflammatory process after mast cell degranulation. PMID- 10390421 TI - Antigen-induced airway inflammation in atopic subjects generates dysfunction of pulmonary surfactant. AB - If pulmonary surfactant develops a dysfunction, its ability to maintain patency of narrow conducting airways diminishes, which is likely to cause an increased airway resistance. We hypothesized that antigen challenge will cause inflammation in the conducting airways and that this will cause a surfactant dysfunction. Twenty atopic patients underwent bronchoalveolar lavage (BAL) 5 min and 48 h after challenge with antigen in one segment and with saline solution in another. BAL fluid (BALF) cell count, differential, and proteins were determined. Surfactant function was studied with a capillary surfactometer (CS), an instrument specifically designed to evaluate surfactant's ability to maintain patency. Eosinophils increased 80-fold 48 h after antigen challenge and total protein increased from 84 to 241 micrograms/ml (median values). BALF surfactant lost part of its ability to maintain openness of the capillary, from 68.8% to 14.0% (p < 0.05). Protein concentration negatively correlated with percent openness (r = -0.62, p = 0.005). We conclude that the antigen challenge resulted in an inflammatory reaction that caused pulmonary surfactant to lose some of its ability to maintain airway patency and speculate that surfactant dysfunction is probably an important factor contributing to increased airway obstruction in allergen-induced exacerbation of asthma. PMID- 10390422 TI - No association between atopy/asthma and the ILe50Val polymorphism of IL-4 receptor. AB - Susceptibility to atopic diseases is known to involve genetic factors. The interleukin-4 (IL-4) receptor- alpha gene (IL4R) reportedly is involved in the development of atopy. A recent study has shown the Ile50 allele of a polymorphism (Ile50Val) of IL4R to be associated with atopy. The objective of this study was to replicate this association and confirm the possible role of the Ile50Val polymorphism of IL4R in the etiology of atopic asthma in a Japanese population. We conducted a transmission disequilibrium test in 86 families identified through asthmatic children. A case-control study was also carried out using both atopic and control subjects. The IL4R Ile50Val polymorphism was genotyped by a PCR restriction fragment length polymorphism method using an intronic upstream primer. The IL4R Ile50 allele was not preferentially transmitted to atopy- or to asthma-affected children. Neither the Ile50 allele nor the Ile50/Ile50 genotype was more prevalent in the atopic subjects than in the control subjects. Our findings indicate that the Ile50Val polymorphism of IL4R does not play a substantial role in genetic predisposition for the etiology of atopy or asthma in this Japanese population. PMID- 10390423 TI - Initial microbiologic studies did not affect outcome in adults hospitalized with community-acquired pneumonia. AB - Microbiologic studies (MBSs) fail to identify a specific pathogen in more than 50% of patients with community-acquired pneumonia (CAP). The 1993 American Thoracic Society guideline (ATS-GL) for the management of CAP advised selecting initial antibiotic regimens based on severity of illness and comorbidities. Our study evaluated the role of initial MBS in adult patients hospitalized with CAP and treated according to the ATS-GL. In 184 patients hospitalized at our facility for CAP in 1996, and treated according to the ATS-GL, 25 (14%) failed to respond to initial antibiotic regimens. In these nonresponders, there was no difference in mortality between those in whom antibiotics were changed empirically, and those with MBS-guided changes. We conclude that initial MBS may not be warranted in many adult patients admitted for CAP. Exceptions include patients with conditions that predispose to less common, more resistant pathogens. PMID- 10390424 TI - Acute purulent exacerbation of chronic obstructive pulmonary disease and Chlamydia pneumoniae infection. AB - In order to investigate the role of bacteria, including Mycoplasma pneumoniae and especially Chlamydia pneumoniae in acute purulent exacerbations of chronic obstructive pulmonary disease (COPD), we examined sputum specimens and acute and convalescent sera taken 26 d apart from 49 outpatients experiencing an acute purulent exacerbation of COPD. The sera were tested for antibodies to C. pneumoniae with the microimmunofluorescence test, and for antibodies to M. pneumoniae with the indirect fluorescence antibody test. Routine microbiologic culture of sputum yielded potentially pathogenic microorganisms in 12 of the 49 patients (24%). Three patients (6%) showed serologic evidence of recent M. pneumoniae infection. Seven patients showed high IgG titers of >/= 1:1,024 to C. pneumoniae, and an additional four had a fourfold increase in IgG titer, suggesting reinfection with C. pneumoniae. Sputum from two of these 11 patients also grew Streptococcus pneumoniae, and one grew Moraxella catarrhalis. Patients with and without serologic evidence of current C. pneumoniae infection showed no significant differences in clinical features or pulmonary function. The high incidence of infection with C. pneumoniae (the sole causal agent in 16% of cases, and the causal agent with other agents in 6%) provides insight into the importance of this organism among agents leading to exacerbations of COPD in Turkey. PMID- 10390425 TI - Protection against methacholine bronchoconstriction to assess relative potency of inhaled beta2-agonist. AB - The purposes of this study were to estimate the relative dose potency (RP) of two formulations of salbutamol pressurized metered-dose inhalers (Proventil-HFA and Ventolin-CFC MDIs) to protect against methacholine bronchoconstriction, to validate this method and provide recommendations. The protective effects of 100-, 200-, and 400-micrograms doses of Proventil-HFA were compared with the same doses of Ventolin-CFC in 18 adult asthmatics (mean FEV1, 92% predicted; mean baseline PC20 methacholine, 1.8 mg/ml), in a dose-level blind, balanced, eight-period, crossover, placebo-controlled study. The log-transformed PC20 values after each dose of the drugs were compared by repeated-measures analysis of variance (ANOVA). A significant dose-effect was present (p < 0.0001). Using the Finney assay, the RP of Proventil-HFA compared with Ventolin-CFC was 1.08 (90% CI, 0.81 1.46) (80% power). This was also estimated using a nonlinear Emax model to validate the Finney method. The most precise estimate of RP was obtained with the comparison between 100- and 200-micrograms doses (RP, 1.00; 90% CI, 0.77-1.31). There were no adverse events resulting from the drugs or methacholine. We conclude that Proventil-HFA salbutamol is bioequivalent to Ventolin-CFC salbutamol. Bronchoprotection to methacholine is a valid method of demonstrating bioequivalence. By this method, 100- and 200-micrograms doses of salbutamol inhalations from an MDI will suffice. PMID- 10390426 TI - Outcomes research in critical care: results of the American Thoracic Society Critical Care Assembly Workshop on Outcomes Research. The Members of the Outcomes Research Workshop. PMID- 10390427 TI - Idiopathic congenital central hypoventilation syndrome: diagnosis and management. American Thoracic Society. PMID- 10390428 TI - Nosocomial Acquisition of Burkholderia gladioli in patients with cystic fibrosis. PMID- 10390429 TI - Managing acute anaphylaxis. New guidelines emphasise importance of intramuscular adrenaline. PMID- 10390430 TI - PFI: perfidious financial idiocy. A "free lunch" that could destroy the NHS. PMID- 10390431 TI - Magnesium sulphate and pre-eclampsia. Trial needed to see whether it's as valuable in pre-eclampsia as in eclampsia. PMID- 10390432 TI - A little bit of measles does you good. Even if measles is eradicated, immunisation may still be desirable in developing countries. PMID- 10390433 TI - Evaluating NHS direct. Early findings raise questions about expanding the service. PMID- 10390434 TI - UK introduces far reaching law to protect whistleblowers. PMID- 10390435 TI - Antihistamine drug withdrawn by manufacturer. PMID- 10390436 TI - UK government confirms ban on human reproductive cloning. PMID- 10390437 TI - United States clears silicone breast implants. PMID- 10390438 TI - In brief PMID- 10390439 TI - Surgeons develop composite bone transplants. PMID- 10390441 TI - BUPA moves towards managed care PMID- 10390440 TI - Scanning shows structural abnormality in headache PMID- 10390442 TI - Link between magnetic fields and leukaemia is weak PMID- 10390444 TI - Some operating theatres used only half time PMID- 10390443 TI - NHS librarians cannot access the internet. PMID- 10390445 TI - Donations of useless medicines to Kosovo contributes to chaos. PMID- 10390447 TI - Spain aims to slow the rise in spending on drugs. PMID- 10390448 TI - Royal college demands 2000 more NHS consultant physicians PMID- 10390446 TI - EU adopts tougher safeguards on genetic modification. PMID- 10390449 TI - UK GPs will produce blueprint for the future PMID- 10390450 TI - Task force for junior doctors' hours in wales PMID- 10390451 TI - Outcome of long stay psychiatric patients resettled in the community: prospective cohort study. AB - OBJECTIVE: To examine the outcome of a population of long stay psychiatric patients resettled in the community. DESIGN: Prospective study with 5 year follow up. SETTING: Over 140 residential settings in north London. SUBJECTS: 670 long stay patients from two London hospitals (Friern and Claybury) discharged to the community from 1985 to 1993. MAIN OUTCOME MEASURES: Continuity and quality of residential care, readmission to hospital, mortality, crime, and vagrancy. RESULTS: Of the 523 patients who survived the 5 year follow up period, 469 (89.6%) were living in the community by the end of follow up, 310 (59.2%) in their original community placement. A third (210) of all patients were readmitted at least once. Crime and homelessness presented few problems. Standardised mortality ratios for the group were comparable with those reported for similar populations. CONCLUSIONS: When carefully planned and adequately resourced, community care for long stay psychiatric patients is beneficial to most individuals and has minimal detrimental effects on society. PMID- 10390452 TI - Serious hazards of transfusion (SHOT) initiative: analysis of the first two annual reports. AB - OBJECTIVE: To receive and collate reports of death or major complications of transfusion of blood or components. DESIGN: Haematologists were invited confidentially to report deaths and major complications after blood transfusion during October 1996 to September 1998. SETTING: Hospitals in United Kingdom and Ireland. SUBJECTS: Patients who died or experienced serious complications, as defined below, associated with transfusion of red cells, platelets, fresh frozen plasma, or cryoprecipitate. MAIN OUTCOME MEASURES: Death, "wrong" blood transfused to patient, acute and delayed transfusion reactions, transfusion related acute lung injury, transfusion associated graft versus host disease, post transfusion purpura, and infection transmitted by transfusion. Circumstances relating to these cases and relative frequency of complications. RESULTS: Over 24 months, 366 cases were reported, of which 191 (52%) were "wrong blood to patient" episodes. Analysis of these revealed multiple errors of identification, often beginning when blood was collected from the blood bank. There were 22 deaths from all causes, including three from ABO incompatibility. There were 12 infections: four bacterial (one fatal), seven viral, and one fatal case of malaria. During the second 12 months, 164/424 hospitals (39%) submitted a "nil to report" return. CONCLUSIONS: Transfusion is now extremely safe, but vigilance is needed to ensure correct identification of blood and patient. Staff education should include awareness of ABO incompatibility and bacterial contamination as causes of life threatening reactions to blood. PMID- 10390453 TI - Cost minimisation analysis of provision of oxygen at home: are the drug tariff guidelines cost effective? AB - OBJECTIVES: To determine the level of oxygen cylinder use at which it becomes more cost effective to provide oxygen by concentrator at home in Northern Ireland, and to examine potential cost savings if cylinder use above this level had been replaced by concentrator in 1996. DESIGN: Cost minimisation analysis. SETTING: Area health boards in Northern Ireland. MAIN OUTCOME MEASURES: Cost effective cut off point for switch to provision of oxygen from cylinder to concentrator. Potential maximum and minimum savings in Northern Ireland (sensitivity analysis) owing to switch to more cost effective strategy on the basis of provision of cylinders in 1996. RESULTS: In Northern Ireland it is currently cost effective to provide oxygen by concentrator when the patient is using three or more cylinders per month independent of the duration of the prescription. More widespread use of concentrators at this level of provision is likely to lead to a cost saving. CONCLUSIONS: The Drug Tariff prescribing guidelines, advocating that provision of oxygen by concentrator becomes cheaper when 21 cylinders are being used per month-are currently inaccurate in Northern Ireland. Regional health authorities should review their current arrangements for provision of oxygen at home and perform a cost analysis to determine at what level it becomes more cost effective to provide oxygen by concentrator. PMID- 10390454 TI - Risk of HIV related Kaposi's sarcoma and non-Hodgkin's lymphoma with potent antiretroviral therapy: prospective cohort study. Swiss HIV Cohort Study. PMID- 10390455 TI - Randomised controlled trial of long term efficacy of captopril on preservation of kidney function in normotensive patients with insulin dependent diabetes and microalbuminuria. PMID- 10390456 TI - Socioeconomic differences in general practice consultation rates in patients aged 65 and over: prospective cohort study. AB - OBJECTIVE: To examine socioeconomic differences in general practice consultation rates among patients aged 65 years and over. DESIGN: Secondary analysis of data from the fourth national survey of morbidity in general practice. SETTING: 60 general practices in England and Wales. SUBJECTS: 71 984 people aged 65 years and over. MAIN OUTCOME MEASURES: Annual contact rates and home visiting rates with general practitioners and practice nurses. RESULTS: Social class differences in tact rates were greatest in 65-74 year olds, with rates 23% higher in patients from social class V than in class I (4.82 v 3.93 per person). In 75-84 year olds there was no clear association between social class and contact rates, and in people aged >/=85 years contact rates were highest in patients from class I. Home visiting rates were twice as high in patients from class V as in patients from class I (1.38 v 0.66 per person). Contact rates were 17% higher in people living in communal establishments and 8% higher in those living alone than in those living with others but not in a communal establishment. 66% of contacts with patients in communal establishments and 26% of those with patients living alone were in patients' homes compared with 18% with those living in standard accommodation. These differences persisted after adjustment in a generalised linear model. CONCLUSIONS: Elderly people show socioeconomic differences in consultation rates. The extra workload generated by elderly people living alone and in communal establishments suggests additional payments to general practitioners are needed. PMID- 10390457 TI - Health information and interaction on the internet: a survey of female urinary incontinence. AB - OBJECTIVE: To evaluate the internet as a source of information about urinary incontinence and to explore interactive facilities. DESIGNLimited survey of internet resources. SUBJECTS: 75 websites providing information about incontinence and an opportunity for interactivity, 25 web doctors, and two news groups. MAIN OUTCOME MEASURES: Quality scores according to predefined general and specific criteria. Internet popularity indexes according to number of links to websites. Correlation between quality scores and popularity indexes. RESULTS: Few sites provided comprehensive information, but the information actually provided was mostly correct. Internet popularity indexes did not correlate with quality scores. The most informative site was easily found with general internet search engines but was not found in any of the medical index sites investigated. Sixty six per cent of sites responded to an email request for advice from a fictitious incontinent woman, half of them within 24 hours. Twelve responders provided vital information that the woman might suffer from drug induced incontinence. CONCLUSIONS: Excellent information about urinary incontinence was found on the internet, but the number of links to a site did not reflect quality of content. Patients may get valuable advice and comfort from using interactive services. PMID- 10390458 TI - Computer support for recording and interpreting family histories of breast and ovarian cancer in primary care (RAGs): qualitative evaluation with simulated patients. AB - OBJECTIVES: To explore general practitioners' attitudes towards and use of a computer program for assessing genetic risk of cancer in primary care. DESIGN: Qualitative analysis of semistructured interviews and video recordings of simulated consultations. PARTICIPANTS: Purposive sample of 15 general practitioners covering a range of computer literacy, interest in genetics, age, and sex. INTERVENTIONS: Each doctor used the program in two consultations in which an actor played a woman concerned about her family history of cancer. Consultations were videotaped and followed by interviews with the video as a prompt to questioning. MAIN OUTCOME MESURESs: Use of computer program in the Consultation. RESULTS: The program was viewed as an appropriate application of information technology because of the complexity of cancer genetics and a sense of "guideline chaos" in primary care. Doctors found the program easy to use, but it often affected their control of the consultation. They needed to balance their desire to share the computer screen with the patient, driven by their concerns about the effect of the computer on doctor-patient communication, against the risk of premature disclosure of bad news. CONCLUSIONS: This computer program could provide the necessary support to assist assessment of genetic risk of cancer in primary care. The potential impact of computer software on the consultation should not be underestimated. This study highlights the need for careful evaluation when developing medical information systems. PMID- 10390459 TI - From cerebral malaria to preventive medicine PMID- 10390461 TI - Trains, worry, and disease PMID- 10390460 TI - Science, medicine, and the future. Behaviour and genes. PMID- 10390462 TI - ABC of intensive care. Renal support. PMID- 10390463 TI - A spirit of adventure prevails PMID- 10390464 TI - Diurnal variability--time to change asthma guidelines? PMID- 10390465 TI - The private finance initiative. NHS capital expenditure and the private finance initiative-expansion or contraction? PMID- 10390466 TI - Children born after intracytoplasmic sperm injection. What do authors mean by control infants? PMID- 10390467 TI - UK inquiry should establish why contaminated blood products were given to people with haemophilia. PMID- 10390468 TI - Proposed appraisal system and political correctness. Consultant appraisals will be useful. PMID- 10390469 TI - Combining the two neonatal examinations. In primary care, second examination is useful. PMID- 10390470 TI - GMC's current proposals for revalidation are flawed. PMID- 10390471 TI - Cyclosporin neurotoxicity after chemotherapy. Case had features of thrombotic thrombocytopenic purpura and haemolytic uraemic syndrome. PMID- 10390472 TI - Devolution in Latin America has had poor effects on health care. PMID- 10390473 TI - Scientific foundation of mammographic screening is based on inconclusive research in Sweden. PMID- 10390474 TI - Funding long term care for older people. Heirs will have to forgo their inheritance. PMID- 10390475 TI - Magnetic resonance necropsy is offered routinely in university college London hospitals. PMID- 10390476 TI - Arguments in editorial were not "biologically implausible". PMID- 10390477 TI - Surely doctors should follow the spirit, rather than the letter, of the law. PMID- 10390478 TI - Patients were more satisfied with chiropractic than other treatments for low back pain. PMID- 10390479 TI - Surveillance of congenital rubella in Great Britain. Rubella can be mistaken for parvovirus b19 infection. PMID- 10390480 TI - Withdrawing low risk women from cervical screening programmes. Conclusions cannot yet be drawn. PMID- 10390481 TI - Frequent changes in policy risk confusion among health workers. PMID- 10390482 TI - High security facilities must not be closed until viable alternatives are in place. PMID- 10390483 TI - Annual general meeting of the BMA. Bma or UKMA? PMID- 10390484 TI - Iceland's medical database is insecure. PMID- 10390485 TI - Sir ian fraser PMID- 10390486 TI - Local medical committee conference PMID- 10390487 TI - Demanding medical excellence PMID- 10390488 TI - On the history of lunacy: the 19th century and after PMID- 10390490 TI - Starting back at the bottom PMID- 10390489 TI - Doctors as gods. PMID- 10390492 TI - Getting things wrong PMID- 10390491 TI - Beastly handwriting PMID- 10390493 TI - Community care of long stay psychiatric patients presents few problems PMID- 10390494 TI - Over half of cases of transfusion error involve giving the wrong blood PMID- 10390495 TI - Provision of oxygen at home in northern ireland is not cost effective PMID- 10390496 TI - Antiretroviral therapy does not prevent HIV related non-Hodgkin's lymphoma PMID- 10390497 TI - Social class affects old people's use of general practice PMID- 10390500 TI - Retirement PMID- 10390499 TI - Dr. Makoto hayashi PMID- 10390498 TI - Computer program may help in assessing genetic risk of cancer in primary care PMID- 10390501 TI - Evaluation of transcriptional fusions with green fluorescent protein versus luciferase as reporters in bacterial mutagenicity tests. AB - A bacterial plasmid was constructed on which the regulatory region of the umuC gene of Escherichia coli was fused to the coding sequence of the green fluorescent protein gene (gfp) from the jellyfish Aequorea victoria. Escherichia coli AB1157 strains carrying the plasmid emitted fluorescence in the presence of mutagens that induce the SOS DNA repair system. Data on tests with nitrosoguanidine, methylmethane sulphonate and UV radiation (254 nm) are presented. Although fluorescent detection using this system was not as rapid or sensitive as a similar luminescent equivalent (umuC-luxAB), the gfp reporter system was more robust. Escherichia coli umu gene induction was also analysed in Salmonella typhimurium TA1537 cells following plasmid transfer and exposure to the same range of mutagens. There was no significant difference in sensitivity between the two species. These preliminary results will provide the basis for development of mutagenicity test systems useful in the testing of complex mixtures, such as environmental samples, and the investigation of physiological parameters influencing spontaneous mutagenesis in bacteria. PMID- 10390502 TI - Analysis of bleomycin-induced chromosomal aberrations in Chinese hamster primary embryonic cells by FISH using arm-specific painting probes. AB - Chinese hamster primary embryonic cells (at G1 phase) were treated with 1.0 or 3.0 microg/ml bleomycin and chromosomal aberrations in first division metaphases were analysed by fluorescence in situ hybridization (FISH) using arm-specific painting probes for chromosomes 3, 4, 8 and 9. We observed that bleomycin induced all classes of chromosome-type aberrations very efficiently. The interesting findings were: (i) the frequency of induced interstitial translocations (i.e. insertions) was approximately equal to that of reciprocal translocations; (ii) the frequency of induced pericentric inversions was higher than that of centric rings. In our earlier studies, we found that X-rays induced a low frequency of interstitial translocations in comparison with reciprocal translocations and equal frequencies of centric rings and pericentric inversions. These data suggest that bleomycin differs from X-rays with respect to the induction of some specific types of aberrations. The results of a chi2 test examining the hypothesis that formed aberrations among the chromosomes or chromosome arms are randomly distributed on the basis of their relative lengths revealed a differential involvement of these chromosomes in the aberrations following exposure to bleomycin. In general, chromosome 8 was found to be more involved in induced aberrations than expected, chromosome 4 was randomly involved, whereas chromosomes 3 and 9 were less involved. This study demonstrates the utility of arm-specific painting probes for efficient detection of a large variety of chromosomal aberrations induced by bleomycin. PMID- 10390503 TI - Cytosol is required for the modulation by dietary casein of the hepatic microsomal activation of aflatoxin B1 to mutagenic metabolites detectable in Salmonella. AB - We have shown previously that dietary protein (casein) levels can affect the ability of rat liver S9 to metabolize aflatoxin B1 (AFB) as well as other promutagens detectable in Salmonella strain TA98 [Mutat. Res. (1997), 360, 115 126 and 127-143]. The mutagenic potency of AFB was greatest when metabolized by the Aroclor 1254-induced hepatic S9 prepared from F344 male rats that consumed an isocaloric, semisynthetic diet for 6 weeks that contained an adequate (12%) level of methionine-supplemented casein as the sole protein source, compared with S9s from rats fed diets that contained nominally deficient (8%) or high (22%) levels of casein. Here we have extended this observation by performing (i) mutagenicity studies with microsomes, cytosols and reconstituted S9s (recombinations of microsomes and cytosols across dietary groups), and (ii) in vitro incubations followed by analysis of metabolites by fluorescence high-pressure liquid chromatography. Microsomes, but not cytosols, activated AFB; however, activation to the level observed with S9 occurred only when microsomes from the rats fed 12% casein were combined with cytosols from any dietary group. Consistent with the mutagenicity results, the greatest metabolism of the AFB parent compound and the highest level of the glutathione conjugate of the presumptively identified AFB exo-8,9-epoxide (the ultimate mutagenic form of AFB) were produced by S9s from the rats fed the 12% casein diet. The levels of these metabolites and the mutagenicity of AFB changed in parallel with changes in dietary casein levels. In summary, cytosolic elements, which are not affected by dietary casein levels, interact with microsomal enzymes, which are modulated by dietary casein levels, to influence the ability of hepatic S9 to activate AFB to a mutagen. PMID- 10390504 TI - Anaphase aberrations in the embryos of the marine tubeworm Pomatoceros lamarckii (Polychaeta: Serpulidae): a new in vivo test assay for detecting aneugens and clastogens in the marine environment. AB - The marine environment receives a wide variety of chemical inputs, many of which have the potential to damage DNA or interfere with the process of cell division. Here we describe a new assay based on the early embryo and larval stages of a planktonic spawning, tube dwelling marine worm, Pomatoceros lamarckii, which for experimental purposes has the advantage of producing large numbers of ripe gametes throughout the majority of the year. One of the most promising end-points is the use of dividing cells to detect anaphase aberrations such as lagging chromosomes, tripolar anaphases, acentric fragments and chromosome bridges. Apart from the reference mutagens mitomycin C and cyclophosphamide and the well documented spindle poison colchicine, we tested the fungicide carbendazim, a primary metabolite of the fungicide benomyl, and thiabendazole, a pesticide and antihelminthic drug; both of which are known to act as aneugens in other test systems. In addition we tested sodium hypochlorite, a widely used oxidizing agent and disinfectant, di-butylphthalate, a commercial plasticizer and suspected aneugen, and sodium chloride, a recognized non-genotoxin. Significant increases in the frequency of anaphase abnormalities occurred with most test compounds at relatively low concentrations, confirming the sensitivity of the new assay. Sodium chloride yielded a negative response except at the highest non-relevant concentrations, where some chromatid stickiness was observed. In addition, the developmental consequences of exposure to these compounds were assessed in 4-8 cell embryos and at 48 h once the embryos had metamorphosed into free swimming larvae. Mitotic inhibition and anaphase aberrations were found to be a more sensitive indicator of genotoxic exposure than larval development, although there was a suggestion of a possible mechanistic link between aneugenicity/clastogenicity and larval fitness. The new test assay provides a rapid and inexpensive method for screening chemicals and effluents destined for release into the marine environment for potential gamete effects. PMID- 10390505 TI - A proposal for a simple way to distinguish aneugens from clastogens in the in vitro micronucleus test. AB - In our previous in vitro micronucleus (MN) study, we showed that aneugens, in addition to inducing micronuclei, induce a higher frequency of polynuclear (PN) and mitotic (M) cells than clastogens. We hypothesized that the frequency of PN and M cells induced can distinguish aneugens from clastogens. To test the hypothesis, we conducted the micronucleus tests with mitomycin C (MMC), N-methyl N'-nitro-N-nitrosoguanidine (MNNG), vincristine (VINC) and diazepam in a Chinese hamster cell line (CHL) and VINC, benzo[a]pyrene (BP) and 7,12 dimethylbenz[a]anthracene (DMBA) in a subclone of V79 cells (V79-MZ). All chemicals increased the frequency of M cells with statistical significance. All chemicals except diazepam increased the frequency of PN cells with statistical significance. Three of the aneugens (VINC, BP and DMBA) induced >/=200 PN cells/1000 cells while the clastogens (MNNG and MMC) induced 100 PN cells at most. All the aneugens but no clastogens significantly increased the frequency of M cells. We propose that micronucleus test-positive chemicals that induce >/=200 PN cells/1000 cells and significantly increase the frequency of M cells are aneugens and those that induce at most 100 PN cells/1000 cells and do not significantly increase the frequency of M cells in our MN test protocol are clastogens. Diazepam, however, did not induce PN cells, although it increased the frequency of M cells dose dependently. We explain this fact in relation to diazepam's mode of action. Our proposal suggests a quick, easy and practical way to distinguish aneugens from clastogens for screening purposes. PMID- 10390506 TI - CREST staining of micronuclei from free-living rodents to detect environmental contamination in situ. AB - In this work immunofluorescent antikinetochore (CREST) staining was used to analyse bone marrow micronuclei (MN) from free-living animals belonging to four different rodent species. Yellow-necked mice (Apodemus flavicollis) and bank voles (Clethrionomys glareolus) were trapped in the Czech Republic, Algerian mice (Mus spretus) in Spain and house mice (Mus musculus domesticus) in Italy. Animals were collected in areas displaying low or high environmental pollution in order to investigate the sensitivity of CREST analysis on bone marrow MN as a biomarker of environmental stress in situ. Differences in total MN frequencies between animals collected in control or contaminated areas were statistically significant for two species, whereas the differences in CREST+ MN were statistically significant for three species. Interestingly, the percentages of CREST+ MN in animals collected in the control areas were very low (3. 2-8.7%), suggesting that activities inducing alterations in the distribution of chromosomes are very rare in natural conditions. The increased frequencies of CREST+ MN observed in areas with high environmental impact indicate that activities producing loss of chromosomes at mitosis may be characteristic of anthropogenic environments such as industrial settlements around petrochemical factories. Our data suggest that the analysis of CREST+ MN may represent a sensitive end-point for the detection of environmental contamination by genotoxic xenobiotics, offering the advantage of providing information on the mechanism of action of environmental contaminants. PMID- 10390508 TI - A micromethod for the in vitro micronucleus assay. AB - A micromethod for the in vitro micronucleus assay was developed using L5178Y cells to enable the rapid screening of a large number of molecules. The method is quick, simple to perform and needs very small amounts of compound, i.e. <10 mg. In this methodology, three types of treatment were carried out in parallel, enabling an optimal detection of both aneugenic and clastogenic compounds: two treatments without metabolic activation with or without a recovery period after a 24 h continuous treatment and one treatment with metabolic activation by Aroclor 1254-induced rat or hamster liver S9 mix. Seventeen known genotoxins (12 clastogens and five aneugens) and seven known non-genotoxins were tested. The in vitro micronucleus micromethod using L5178Y cells exhibited good sensitivity (16 positive/17 known genotoxins tested) and specificity (7 negative/7 known non genotoxins tested) for the 24 test compounds studied with or without metabolic activation. Furthermore, this test showed a good correlation with other in vitro micronucleus tests performed using macromethods with various mammalian cell cultures. We conclude that the in vitro micronucleus micromethod with L5178Y cells could be used in the earliest stages of development of new molecules as a preliminary short-term screening assay before starting regulatory tests. PMID- 10390507 TI - Flow cytometric measurement of micronuclei induced in a permanent fish cell line as a possible screening test for the genotoxicity of industrial waste waters. AB - An in vitro micronucleus assay using the permanent fish cell line RTG-2 (rainbow trout gonads) was developed to test industrial waste waters for their genotoxic potential. Comparison of flow cytometric measurement and microscopic scoring of micronucleus frequency with the reference chemicals 1,4-butane sultone (0.2-1 mM), ethylmethane sulphonate (2-10 mM), potassium dichromate (20-100 microM) and benzo[a]pyrene (5-25 microM) showed similar dose-effect relationships. Thirty eight industrial waste waters from 11 different branches of industry obtained from the Bavarian state office for water research were tested using the flow cytometric method (18 from metal processing, 10 from combined waste water, two from synthetic fibre production, one sample each from settlement wastes, non-iron metal manufacturing, leather production, sulphuric acid production, ore processing, graphite film production, cellulose production and flue gas washing). Fourteen of them showed a significant increase in micronucleus frequency. PMID- 10390509 TI - Development of a new bioluminescent mutagenicity assay based on the Ames test. AB - A newly developed rapid mutagenicity assay based on the adenosine triphosphate (ATP)-bioluminescence technique and the Ames test is described. Salmonella typhimurium strains TA98 and TA100 were exposed in an appropriate liquid medium to the direct mutagens 4-nitroquinoline-N-oxide and methyl methanesulphonate, respectively, and to the indirect mutagen 2-aminoanthracene. Both auxotrophic and prototrophic growth were monitored throughout the incubation period as variations in the intracellular ATP levels by means of the luciferin-luciferase assay. After 9-12 h of incubation a dose-response increase in the levels of ATP was readily detected. In order to demonstrate that this increase was due to the growth of revertant bacteria, aliquots from each culture were plated on minimal agar plates. A very good correlation between the changes in ATP levels and the appearance of revertant colonies on the plates was found. Given the rapidity of this method as compared with conventional mutagenicity assays, it has potential for industrial and environmental applications. Other potential applications are also discussed. PMID- 10390510 TI - The application of comparative genomic hybridization and fluorescence in situ hybridization to the characterization of genotoxicity screening tester strains AHH-1 and MCL-5. AB - AHH-1 TK+/- is a human B cell-derived lymphoblastoid cell line that constitutively expresses a high level of the cytochrome CYP1A1. The MCL-5 cell line was developed by transfection of AHH-1 with cDNAs encoding the human cytochrome P450s, CYP1A2, CYP2A6, CYP2E1, CYP3A4 and microsomal epoxide hydrolase carried in plasmids. The metabolic components of these cell lines make them a useful screening tool for use in mutagenicity studies. Although AHH-1 and MCL-5 are closely related, the two cell lines show differences which cannot be attributed to transfection. In the present study both cell lines were investigated for chromosome stability by comparative genomic hybridization (CGH) and fluorescence in situ hybridization (FISH) using whole chromosome probes and telomeric probes. Amplification in chromosomes 4q, 3q and 9p was observed in both cell lines. To compare the cell lines directly, AHH-1 and MCL-5 DNAs were co hybridized on the same metaphases using a modified CGH technique. The only difference observed between AHH-1 and MCL-5 was the degree of amplification involving the subtelomeric region of chromosome 4; the additional telomeric region (4q) was translocated onto chromosome 11 and/or chromosome X. FISH was use to show the presence of isochromosomes 3q and 9p in both cell lines with a chromosome number of 48 or higher. These data demonstrate that CGH and FISH with chromosome-specific probes are able to resolve complex karyotypes and to highlight subchromosomal regions involved in rearrangements and potential chromosome fragile sites. Analyses such as those described here may be of considerable value in the determination of the stability of a variety of the cell lines used in the mutagenicity testing of chemicals. PMID- 10390511 TI - Change in centromeric and acentromeric micronucleus frequencies in human populations after chronic radiation exposure. AB - Acute radiation exposure of humans was observed to induce various forms of cytogenetic damage, including increased frequencies of micronuclei and chromosomal aberrations. However, the cytogenetic effects of chronic low dose radiation exposure in vivo needs further characterization. Sixteen subjects with chronic low dose rates of gamma-radiation exposure from 60Co-contaminated steel in radioactive buildings were compared with seven non-exposed reference subjects for micronucleus frequencies after they relocated. By in situ hybridization using a digoxigenin-labeled anti-alpha all human centromere probe, the exposed subjects were shown to have a significant increase in cytochalasin B-modulated micronucleus (CBMN) frequencies, as well as a significant increase in centromere positive (C+) CBMN, centromere-negative (C-) CBMN, total C+signals, single C+ MN signals and multiple C+ signals/1000 binucleated cells (BN). However, decreases in the ratios C+MN/C- MN and C+MN/total CBMN (%) were also noted in the exposed subjects. By mixed effects analysis, considering individuals from the same families, the C- MN and single C+ MN/1000 BN were both positively and moderately associated with previous cumulative exposure. When the time period of relocation post-exposure (relocation time or RT) was considered, total C+MN and multiple C+MN/1000 BN were negatively and significantly associated with RT. Moreover, the C+MN, C- MN, C+MN/C- MN ratio and single C+MN/1000 BN were all negatively and moderately associated with RT, but not with exposure dose. This suggested that acentromeric and single or multiple centromeric CBMN cytogenetic damage seems to disappear differentially in human subjects post chronic low dose radiation exposure. PMID- 10390512 TI - Induction and prevention of micronuclei and chromosomal aberrations in cultured human lymphocytes exposed to the light of halogen tungsten lamps. AB - Previous studies have shown that the light emitted by halogen tungsten lamps contains UV radiation in the UV-A, UV-B and UV-C regions, induces mutations and irreparable DNA damage in bacteria, enhances the frequency of micronuclei in cultured human lymphocytes and is potently carcinogenic to the skin of hairless mice. The present study showed that the light emitted by an uncovered, traditional halogen lamp induces a significant, dose-related and time-related increase not only in micronuclei but also in chromosome-type aberrations, such as breaks, and even more in chromatid-type aberrations, such as isochromatid breaks, exchanges and isochromatid/chromatid interchanges, all including gaps or not, in cultured human lymphocytes. All these genotoxic effects were completely prevented by shielding the same lamp with a silica glass cover, blocking UV radiation. A new model of halogen lamp, having the quartz bulb treated in order to reduce the output of UV radiation, was considerably less genotoxic than the uncovered halogen lamp, yet induction of chromosomal alterations was observed at high illuminance levels. PMID- 10390513 TI - Modification of the Comet assay for the detection of DNA strand breaks in extremely small tissue samples. AB - We modified the Comet assay to enable the quantification of DNA strand breakage in individual cells of extremely small tissue samples. This modification was used to analyze cells isolated from the ectoplacental cone and egg cylinder of mouse embryos at embryonic day 7.5. We detected more naturally occurring DNA strand breaks and a higher number of apoptotic cells in the ectoplacental cone compared with the egg cylinder. PMID- 10390514 TI - Development of cardiac sensitivity to oxygen deficiency: comparative and ontogenetic aspects. AB - Hypoxic states of the cardiovascular system are undoubtedly associated with the most frequent diseases of modern times. They originate as a result of disproportion between the amount of oxygen supplied to the cardiac cell and the amount actually required by the cell. The degree of hypoxic injury depends not only on the intensity and duration of the hypoxic stimulus, but also on the level of cardiac tolerance to oxygen deprivation. This variable changes significantly during phylogenetic and ontogenetic development. The heart of an adult poikilotherm is significantly more resistant as compared with that of the homeotherms. Similarly, the immature homeothermic heart is more resistant than the adult, possibly as a consequence of its greater capability for anaerobic glycolysis. Tolerance of the adult myocardium to oxygen deprivation may be increased by pharmacological intervention, adaptation to chronic hypoxia, or preconditioning. Because the immature heart is significantly more dependent on transsarcolemmal calcium entry to support contraction, the pharmacological protection achieved with drugs that interfere with calcium handling is markedly altered. Developing hearts demonstrated a greater sensitivity to calcium channel antagonists; a dose that induces only a small negative inotropic effect in adult rats stops the neonatal heart completely. Adaptation to chronic hypoxia results in similarly enhanced cardiac resistance in animals exposed to hypoxia either immediately after birth or in adulthood. Moreover, decreasing tolerance to ischemia during early postnatal life is counteracted by the development of endogenous protection; preconditioning failed to improve ischemic tolerance just after birth, but it developed during the early postnatal period. Basic knowledge of the possible improvements of immature heart tolerance to oxygen deprivation may contribute to the design of therapeutic strategies for both pediatric cardiology and cardiac surgery. PMID- 10390515 TI - Apocalmodulin. AB - Intracellular Ca2+ is normally maintained at submicromolar levels but increases during many forms of cellular stimulation. This increased Ca2+ binds to receptor proteins such as calmodulin (CaM) and alters the cell's metabolism and physiology. Calcium-CaM binds to target proteins and alters their function in such a way as to transduce the Ca2+ signal. Calcium-free or apocalmodulin (ApoCaM) binds to other proteins and has other specific effects. Apocalmodulin has roles in the cell that apparently do not require the ability to bind Ca2+ at all, and these roles appear to be essential for life. Apocalmodulin differs from Ca2+-CaM in its tertiary structure. It binds target proteins differently, utilizing different binding motifs such as the IQ motif and noncontiguous binding sites. Other kinds of binding potentially await discovery. The ApoCaM-binding proteins are a diverse group of at least 15 proteins including enzymes, actin binding proteins, as well as cytoskeletal and other membrane proteins, including receptors and ion channels. Much of the cellular CaM is bound in a Ca2+ independent manner to membrane structures within the cell, and the proportion bound changes with cell growth and density, suggesting it may be a storage form. Apocalmodulin remains tightly bound to other proteins as subunits and probably hastens the response of these proteins to Ca2+. The overall picture that emerges is that CaM cycles between its Ca2+-bound and Ca2+-free states and in each state binds to different proteins and performs essential functions. Although much of the research focus has been on the roles of Ca2+-CaM, the roles of ApoCaM are equally vital but less well understood. PMID- 10390516 TI - Signal transduction from the endoplasmic reticulum to the cell nucleus. AB - The endoplasmic reticulum (ER) serves several important functions. Cholesterol, an essential component of cellular membranes, is synthesized on the ER surface. Inside the organelle, proteins destined for secretion or transport to the cell surface are folded and become glycosylated. Because these processes are essential for cell viability, a disturbance in ER function presents significant stress to the cell. In response to ER stress, three distinct signal transduction pathways can be activated. Two of these, the unfolded protein response and the ER-overload response, respond to disturbances in protein processing. The third, the sterol regulatory cascade, is activated by depletion of cholesterol. This review summarizes the recent advances in our understanding of these ER-nuclear signal transduction pathways. In addition, it points to novel regulatory mechanisms discovered in these pathways, which may be widely used in other systems. PMID- 10390517 TI - Microvascular permeability. AB - This review addresses classical questions concerning microvascular permeabiltiy in the light of recent experimental work on intact microvascular beds, single perfused microvessels, and endothelial cell cultures. Analyses, based on ultrastructural data from serial sections of the clefts between the endothelial cells of microvessels with continuous walls, conform to the hypothesis that different permeabilities to water and small hydrophilic solutes in microvessels of different tissues can be accounted for by tortuous three-dimensional pathways that pass through breaks in the junctional strands. A fiber matrix ultrafilter at the luminal entrance to the clefts is essential if microvascular walls are to retain their low permeability to macromolecules. Quantitative estimates of exchange through the channels in the endothelial cell membranes suggest that these contribute little to the permeability of most but not all microvessels. The arguments against the convective transport of macromolecules through porous pathways and for the passage of macromolecules by transcytosis via mechanisms linked to the integrity of endothelial vesicles are evaluated. Finally, intracellular signaling mechanisms implicated in transient increases in venular microvessel permeability such as occur in acute inflammation are reviewed in relation to studies of the molecular mechanisms involved in signal transduction in cultured endothelial cells. PMID- 10390518 TI - Sodium/calcium exchange: its physiological implications. AB - The Na+/Ca2+ exchanger, an ion transport protein, is expressed in the plasma membrane (PM) of virtually all animal cells. It extrudes Ca2+ in parallel with the PM ATP-driven Ca2+ pump. As a reversible transporter, it also mediates Ca2+ entry in parallel with various ion channels. The energy for net Ca2+ transport by the Na+/Ca2+ exchanger and its direction depend on the Na+, Ca2+, and K+ gradients across the PM, the membrane potential, and the transport stoichiometry. In most cells, three Na+ are exchanged for one Ca2+. In vertebrate photoreceptors, some neurons, and certain other cells, K+ is transported in the same direction as Ca2+, with a coupling ratio of four Na+ to one Ca2+ plus one K+. The exchanger kinetics are affected by nontransported Ca2+, Na+, protons, ATP, and diverse other modulators. Five genes that code for the exchangers have been identified in mammals: three in the Na+/Ca2+ exchanger family (NCX1, NCX2, and NCX3) and two in the Na+/Ca2+ plus K+ family (NCKX1 and NCKX2). Genes homologous to NCX1 have been identified in frog, squid, lobster, and Drosophila. In mammals, alternatively spliced variants of NCX1 have been identified; dominant expression of these variants is cell type specific, which suggests that the variations are involved in targeting and/or functional differences. In cardiac myocytes, and probably other cell types, the exchanger serves a housekeeping role by maintaining a low intracellular Ca2+ concentration; its possible role in cardiac excitation-contraction coupling is controversial. Cellular increases in Na+ concentration lead to increases in Ca2+ concentration mediated by the Na+/Ca2+ exchanger; this is important in the therapeutic action of cardiotonic steroids like digitalis. Similarly, alterations of Na+ and Ca2+ apparently modulate basolateral K+ conductance in some epithelia, signaling in some special sense organs (e.g., photoreceptors and olfactory receptors) and Ca2+-dependent secretion in neurons and in many secretory cells. The juxtaposition of PM and sarco(endo)plasmic reticulum membranes may permit the PM Na+/Ca2+ exchanger to regulate sarco(endo)plasmic reticulum Ca2+ stores and influence cellular Ca2+ signaling. PMID- 10390519 TI - Central control of the cardiovascular and respiratory systems and their interactions in vertebrates. AB - This review explores the fundamental neuranatomical and functional bases for integration of the respiratory and cardiovascular systems in vertebrates and traces their evolution through the vertebrate groups, from primarily water breathing fish and larval amphibians to facultative air-breathers such as lungfish and some adult amphibians and finally obligate air-breathers among the reptiles, birds, and mammals. A comparative account of respiratory rhythm generation leads to consideration of the changing roles in cardiorespiratory integration for central and peripheral chemoreceptors and mechanoreceptors and their central projections. We review evidence of a developing role in the control of cardiorespiratory interactions for the partial relocation from the dorsal motor nucleus of the vagus into the nucleus ambiguus of vagal preganglionic neurons, and in particular those innervating the heart, and for the existence of a functional topography of specific groups of sympathetic preganglionic neurons in the spinal cord. Finally, we consider the mechanisms generating temporal modulation of heart rate, vasomotor tone, and control of the airways in mammals; cardiorespiratory synchrony in fish; and integration of the cardiorespiratory system during intermittent breathing in amphibians, reptiles, and diving birds. Concluding comments suggest areas for further productive research. PMID- 10390520 TI - Cardiac ionic currents and acute ischemia: from channels to arrhythmias. AB - The aim of this review is to provide basic information on the electrophysiological changes during acute ischemia and reperfusion from the level of ion channels up to the level of multicellular preparations. After an introduction, section II provides a general description of the ion channels and electrogenic transporters present in the heart, more specifically in the plasma membrane, in intracellular organelles of the sarcoplasmic reticulum and mitochondria, and in the gap junctions. The description is restricted to activation and permeation characterisitics, while modulation is incorporated in section III. This section (ischemic syndromes) describes the biochemical (lipids, radicals, hormones, neurotransmitters, metabolites) and ion concentration changes, the mechanisms involved, and the effect on channels and cells. Section IV (electrical changes and arrhythmias) is subdivided in two parts, with first a description of the electrical changes at the cellular and multicellular level, followed by an analysis of arrhythmias during ischemia and reperfusion. The last short section suggests possible developments in the study of ischemia-related phenomena. PMID- 10390522 TI - Construction of a variability map for eukaryotic large subunit ribosomal RNA. AB - In this paper, we present a variability map of the eukaryotic large subunit ribosomal RNA, showing the distribution of variable and conserved sites in this molecule. The variability of each site in this map is indicated by means of a colored dot. Construction of the variability map was based on the substitution rate calibration (SRC) method, in which the substitution rate of each nucleotide site is computed by looking at the frequency with which sequence pairs differ at that site as a function of their evolutionary distance. Variability maps constructed by this method provide a much more accurate and objective description of site-to-site variability than visual inspection of sequence alignments. PMID- 10390521 TI - Ion channels in presynaptic nerve terminals and control of transmitter release. AB - The primary function of the presynaptic nerve terminal is to release transmitter quanta and thus activate the postsynaptic target cell. In almost every step leading to the release of transmitter quanta, there is a substantial involvement of ion channels. In this review, the multitude of ion channels in the presynaptic terminal are surveyed. There are at least 12 different major categories of ion channels representing several tens of different ion channel types; the number of different ion channel molecules at presynaptic nerve terminals is many hundreds. We describe the different ion channel molecules at the surface membrane and inside the nerve terminal in the context of their possible role in the process of transmitter release. Frequently, a number of different ion channel molecules, with the same basic function, are present at the same nerve terminal. This is especially evident in the cases of calcium channels and potassium channels. This abundance of ion channels allows for a physiological and pharmacological fine tuning of the process of transmitter release and thus of synaptic transmission. PMID- 10390523 TI - A subtelomeric DNA sequence is required for correct processing of the macronuclear DNA sequences during macronuclear development in the hypotrichous ciliate Stylonychia lemnae. AB - During macronuclear differentiation in ciliated protozoa a series of programed DNA reorganization processes occur. These include the elimination of micronuclear specific DNA sequences, the specific fragmentation of the genome into small gene sized DNA molecules, the de novo addition of telomeric sequences to these DNA molecules and the specific amplification of the remaining DNA molecules. Recently we constructed a vector containing the modified micronuclear version of macronuclear destined DNA sequences that was correctly fragmented and telomeres were added de novo after injection into the developing macronucleus. It therefore must contain all the cis- acting sequences required for these processes. We made a series of vectors deleting different sequences from the original vector. It could be shown that at least in the case studied here no micronuclear-specific sequences are required for specific fragmentation of the genome and telomere addition. However, a short subtelomeric sequence at the 3[prime]-end is essential for these processes, whereas no specific cut seems to occur at the 5[prime]-end. In addition, we can show that the processing activity is restricted to a short period of time during macronuclear differentiation and that a preceding transcription is required for correct processing of macronuclear-destined DNA sequences. Possible mechanisms of these processes will be discussed. PMID- 10390524 TI - Codon usage as a tool to predict the cellular location of eukaryotic ribosomal proteins and aminoacyl-tRNA synthetases. AB - In spite of many efforts, the prediction of the location of proteins in eukaryotic cells (cytoplasm, mitochondrion or chloroplast) is still far from straightforward. In some cases (e.g. ribosomal proteins and aminoacyl-tRNA synthetases) both the cytoplasmic proteins and their organellar counterparts are encoded by the nuclear genome. A factorial correspondence analysis of the codon usage in yeast and Caenorhabditis elegans shows that the codon usage of those nuclear genes encoding ribosomal proteins or aminoacyl-tRNA synthetases is markedly different, depending on the final location of the proteins (cytoplasmic or mitochondrial). As a consequence, the location of such proteins-whose sequences are now frequently determined by systematic genomic sequencing-can be easily and quickly predicted. A WWW interface has been developed, aimed at providing a user-friendly tool for codon usage pattern analysis. It is available from http://www.genetique.uvsq.fr/afc.html PMID- 10390525 TI - Cytosine methylation transforms an E2F site in the retinoblastoma gene promoter into a binding site for the general repressor methylcytosine-binding protein 2 (MeCP2). AB - The CpG-rich promoter of the retinoblastoma tumor suppressor gene (Rb-1) is normally unmethylated. However, aberrant methylation of CpG dinucleotides within the Rb-1 promoter has been depicted in certain tumors, which determines transcriptional inactivity of the gene and absence of the pRb retinoblastoma protein. Here we have concentrated on an E2F-binding site in the Rb-1 promoter. We show that the E2F site is required for cell-cycle regulated Rb-1 transcription in non-transformed cells. The function of the E2F site is associated with its ability to interact with several activating factors of the E2F family. In contrast, in vitro methylation of two tandemly arranged CpGs in the E2F recognition site prevents binding by E2F factors, and determines instead the recruitment of the general repressor methylcytosine-binding protein 2 (MeCP2). These results suggest that the interaction of MeCP2 with the methylated version of the E2F site may represent a step towards Rb-1 promoter inactivity in tumor cells. PMID- 10390526 TI - 6-Thioguanine alters the structure and stability of duplex DNA and inhibits quadruplex DNA formation. AB - The ability to chemically synthesize biomolecules has opened up the opportunity to observe changes in structure and activity that occur upon single atom substitution. In favorable cases this can provide information about the roles of individual atoms. The substitution of 6-thioguanine (6SG) for guanine is a potentially very useful single atom substitution as 6SG has optical, photocrosslinking, metal ion binding and other properties of potential utility. In addition, 6-mercaptopurine is a clinically important pro-drug that is activated by conversion into 6SG by cells. The results presented here indicate that the presence of 6SG blocks the formation of quadruplex DNA. The presence of 6SG alters the structure and lowers the thermal stability of duplex DNA, but duplex DNA can be formed in the presence of 6SG. These results indicate that some of the cytotoxic activity of 6SG may be due to disruption of the quadruplex structures formed by telomere and other DNAs. This additional mode of action is consistent with the delayed onset of cytotoxicity. PMID- 10390527 TI - Removal of cyclobutane pyrimidine dimers by the UV damage repair and nucleotide excision repair pathways of Schizosaccharomyces pombe at nucleotide resolution. AB - In Schizosaccharomyces pombe two different repair mechanisms remove UV-induced lesions from DNA, i.e. nucleotide excision repair (NER) and UV damage repair (UVDR). Here, the kinetics of removal of cyclobutane pyrimidine dimers (CPDs) by both pathways is determined at base resolution in the transcribed strand (TS) and the non-transcribed strand (NTS) of the sprpb2 +gene. UVDR does not remove lesions in a strand-specific manner, indicating that UVDR is neither stimulated nor inhibited by RNA polymerase II transcription. In contrast, in a UVDR deficient strain the TS is repaired preferentially. This strong strand bias suggests that in S.pombe, as in other species, NER is coupled to transcription. In repair-proficient S.pombe the TS is repaired very rapidly, as a consequence of two efficiently operating pathways, while the NTS is repaired more slowly, mainly by UVDR. Furthermore, we demonstrate that UVDR is not always faster than NER. PMID- 10390528 TI - Origins of the temperature dependence of hammerhead ribozyme catalysis. AB - The difficulties in interpreting the temperature dependence of protein enzyme reactions are well recognized. Here, the hammerhead ribozyme cleavage was investigated under single-turnover conditions between 0 and 60 degrees C as a model for RNA-catalyzed reactions. Under the adopted conditions, the chemical step appears to be rate-limiting. However, the observed rate of cleavage is affected by pre-catalytic equilibria involving deprotonation of an essential group and binding of at least one low-affinity Mg2+ion. Thus, the apparent entropy and enthalpy of activation include contributions from the temperature dependence of these equilibria, precluding a simple physical interpretation of the observed activation parameters. Similar pre-catalytic equilibria likely contribute to the observed activation parameters for ribozyme reactions in general. The Arrhenius plot for the hammerhead reaction is substantially curved over the temperature range considered, which suggests the occurrence of a conformational change of the ribozyme ground state around physiological temperatures. PMID- 10390529 TI - Terminator element mutations affect both the efficiency and position of RNA polymerase I termination in Schizosaccharomyces pombe. AB - RNA polymerase I transcripts, purified from Schizosaccharomyces pombe cells, terminate at three sites that precede 'Sal box'-like termination element (TE) sequences. Essential features in these elements were investigated by the in vivo expression of targeted mutations. RNA analyses confirmed a functional significance for two of the elements (Boxes 1 and 3), but indicated that the third, less related, sequence (Box 2) does not function as a termination signal. The results further indicated that the most conserved residues in the two active TEs, as well as adjacent regions, are also most critical to function. Furthermore, some mutations in these elements or in immediately flanking sequences affect not only the efficiency of termination, but also alter the position of termination by as much as 35 nt. Since the element is able to influence the site of termination over a surprisingly long stretch of DNA sequence, these observations suggest that the TE does not act simply as a pause element by fixing the termination factor. PMID- 10390530 TI - The Wilms' tumor suppressor gene (wt1) product represses different functional classes of transcriptional activation domains. AB - We have studied the ability of the wt1 tumor suppressor gene product to repress different classes of activation domains previously shown to stimulate the initiation and elongation steps of RNA polymerase II transcription in vivo. Repression assays revealed that WT1 represses all three classes of activation domains: Sp1 and CTF, which stimulate initiation (type I), human immunodeficiency virus type I Tat fused to a DNA-binding domain, which stimulates predominantly elongation (type IIA), and VP16, p53 and E2F1, which stimulate both initiation and elongation (type IIB). WT1 is capable of exerting its repression effect over a significant distance when positioned approximately 1700 bp from the core promoter. Deletion analysis of WT1 indicates that the responsible domain resides within the first 180 N-terminal amino acids of the protein. Nuclear run-ons analyzing the effects of WT1 on initiation of transcription demonstrate inhibition of this process. Our observations imply that WT1 can repress activators that stimulate initiation and/or elongation. PMID- 10390531 TI - Cells from XP-D and XP-D-CS patients exhibit equally inefficient repair of UV induced damage in transcribed genes but different capacity to recover UV inhibited transcription. AB - Xeroderma pigmentosum (XP) is a rare hereditary human disorder clinically associated with severe sun sensitivity and predisposition to skin cancer. Some XP patients also show clinical characteristics of Cockayne syndrome (CS), a disorder associated with defective preferential repair of DNA lesions in transcriptionally active genes. Cells from the two XP-patients who belong to complementation group D and exhibit additional clinical symptoms of CS are strikingly more sensitive to the cytotoxic effects of UV-light than cells from classical XP-D patients. To explain the severe UV-sensitivity it was suggested that XP-D-CS cells have a defect in preferential repair of UV-induced 6-4 photoproducts (6-4PP) in active genes. We investigated the capacity of XP-D and XP-D-CS cells to repair UV induced DNA lesions in the active adenosine deaminase gene (ADA) and in the inactive 754 gene by determining (i) the removal of specific lesions, i.e. cyclobutane pyrimidine dimers (CPD) and 6-4PP, or (ii) the formation of BrdUrd labeled repair patches. No differences in repair capacity were observed between XP-D and XP-D-CS cells. In both cell types repair of CPD was completely absent whereas 6-4PP were inefficiently removed from the ADA gene and the 754 gene with similar kinetics. However, whereas XP-D cells were able to restore UV-inhibited RNA synthesis after a UV-dose of 2 J/m2, RNA synthesis in XP-D-CS cells remained repressed up to 24 h after irradiation. Our results are inconsistent with the hypothesis that differences in the capacity to perform preferential repair of UV induced photolesions in active genes between XP-D and XP-D-CS cells are the cause of the extreme UV-sensitivity of XP-D-CS cells. Rather, the enhanced sensitivity of XP-D-CS cells may be associated with a defect in transcription regulation superimposed on the repair defect. PMID- 10390533 TI - Synthetic substrate analogs for the RNA-editing adenosine deaminase ADAR-2. AB - We have synthesized structural analogs of a natural RNA editing substrate and compared editing reactions of these substrates by recombinant ADAR-2, an RNA editing adenosine deaminase. Deamination rates were shown to be sensitive to structural changes at the 2[prime]-carbon of the edited adenosine. Methylation of the 2[prime]-OH caused a large decrease in deamination rate, whereas 2[prime] deoxyadenosine and 2[prime]-deoxy-2[prime]-fluoroadenosine were deaminated at a rate similar to adenosine. In addition, a duplex containing as few as 19 bp of the stem structure adjacent to the R/G editing site of the GluR-B pre-mRNA supports deamination of the R/G adenosine by ADAR-2. This identification and initial characterization of synthetic RNA editing substrate analogs further defines structural elements in the RNA that are important for the deamination reaction and sets the stage for additional detailed structural, thermodynamic and kinetic studies of the ADAR-2 reaction. PMID- 10390532 TI - Plasmid linking number change induced by topoisomerase I-mediated DNA damage. AB - The state of cellular chromatin in response to DNA damage has been examined by monitoring the change in the linking number of circular episomes. COS cells transfected with an SV40-based vector were treated with camptothecin (CPT), a eukaryotic DNA topoisomerase I (TOP1) poison which induces TOP1-mediated DNA damage. Within minutes, a large increase in the linking number (over 10 linking number) of a small fraction (5-15%) of the episomal DNA was observed. A similar CPT-induced increase in plasmid DNA linking number was observed in Saccharomyces cerevisae expressing human DNA TOP1. In this case, the majority of the plasmid DNA can undergo rapid relaxation. The large increase in the plasmid linking number suggests major chromatin structural reorganization in response to TOP I mediated DNA damage. PMID- 10390534 TI - Formation of DNA adducts by formaldehyde-activated mitoxantrone. AB - Recent studies with the anthracycline Adriamycin have demonstrated its activation by formaldehyde and subsequent binding to DNA in vitro. Since formaldehyde levels are known to be higher in cells of myeloid origin and the structurally related drug mitoxantrone is most effective against cancers of myeloid origin, this indicates a possible role of formaldehyde in the activation of mitoxantrone. In vitro studies revealed that the activation of mitoxantrone by formaldehyde leads to the formation of drug-DNA adducts. These adducts stabilised DNA such that they functioned as virtual interstrand crosslinks. The interstrand crosslinks were formed in the presence of mitoxantrone and formaldehyde in a time- and concentration-dependent manner. In the absence of formaldehyde no crosslinks were formed, indicating a key role in drug activation and DNA binding. The adducts (virtual crosslinks) were relatively unstable with 50% crosslinks remaining after 10 min at 60 degrees C in 45% formamide. Like Adriamycin, the mitoxantrone formaldehyde-DNA crosslinks are heat labile and do not display the stability associated with covalent interstrand crosslinks. PMID- 10390535 TI - A bipartite sequence element associated with matrix/scaffold attachment regions. AB - We have identified a MAR/SAR recognition signature (MRS) which is common to a large group of matrix and scaffold attachment regions. The MRS is composed of two degenerate sequences (AATAAYAA and AWWRTAANNWWGNNNC) within close proximity. Analysis of >300 kb of genomic sequence from a variety of eukaryotic organisms shows that the MRS faithfully predicts 80% of MARs and SARs. In each case where we find a MRS, the corresponding DNA region binds specifically to the nuclear scaffold. Although all MRSs are associated with a SAR, not all known SARs and MARs contain a MRS, suggesting that at least two classes exist, one containing a MRS, the other not. Evidence is presented that the two sequence elements of the bipartite MRS occupy a position on the nucleosome near the dyad axis, together creating a putative protein binding site. The identification of a MAR- and SAR associated DNA element is an important step forward towards understanding the molecular mechanisms of these elements. It will allow: (i) analysis of the genomic location of SARs, e.g. in relationship to genes, based on sequence information alone, rather than on the basis of an elaborate biochemical assay; (ii) identification and analysis of proteins that specifically bind to the MRS. PMID- 10390536 TI - Linkers designed to intercalate the double helix greatly facilitate DNA alkylation by triplex-forming oligonucleotides carrying a cyclopropapyrroloindole reactive moiety. AB - Triplex-forming oligonucleotides (TFOs) bind sequence-specifically in the major groove of double-stranded DNA. Cyclopropapyrroloindole (CPI), the electrophilic moiety that comprises the reactive subunit of the antibiotic CC-1065, gives hybridization-triggered alkylation at the N-3 position of adenines when bound in the minor groove of double-stranded DNA. In order to attain TFO-directed targeting of CPI, we designed and tested linkers to 'thread' DNA from the major groove-bound TFO to the minor groove binding site of CPI. Placement of an aromatic ring in the linker significantly enhanced the site-directed reaction, possibly due to a 'threading' mechanism where the aromatic ring is intercalated. All of the linkers containing aromatic rings provided efficient alkylation of the duplex target. The linker containing an acridine ring system, the strongest intercalator in the series, gave a small but clearly detectable amount of non-TFO specific alkylation. An equivalent-length linker without an aromatic ring was very inefficient in DNA target alkylation. PMID- 10390537 TI - A dimer of the lymphoid protein RAG1 recognizes the recombination signal sequence and the complex stably incorporates the high mobility group protein HMG2. AB - RAG1 and RAG2 are the two lymphoid-specific proteins required for the cleavage of DNA sequences known as the recombination signal sequences (RSSs) flanking V, D or J regions of the antigen-binding genes. Previous studies have shown that RAG1 alone is capable of binding to the RSS, whereas RAG2 only binds as a RAG1/RAG2 complex. We have expressed recombinant core RAG1 (amino acids 384-1008) in Escherichia coli and demonstrated catalytic activity when combined with RAG2. This protein was then used to determine its oligomeric forms and the dissociation constant of binding to the RSS. Electrophoretic mobility shift assays show that up to three oligomeric complexes of core RAG1 form with a single RSS. Core RAG1 was found to exist as a dimer both when free in solution and as the minimal species bound to the RSS. Competition assays show that RAG1 recognizes both the conserved nonamer and heptamer sequences of the RSS. Zinc analysis shows the core to contain two zinc ions. The purified RAG1 protein overexpressed in E.coli exhibited the expected cleavage activity when combined with RAG2 purified from transfected 293T cells. The high mobility group protein HMG2 is stably incorporated into the recombinant RAG1/RSS complex and can increase the affinity of RAG1 for the RSS in the absence of RAG2. PMID- 10390538 TI - A human Raf-responsive zinc-finger protein that binds to divergent sequences. AB - LZ321, a human liver cDNA, encodes a protein that bound to a Drosophila tramtrack binding site, GGTCCT. The sequence of LZ321 matched that of RREB1, a transcription factor that bound to a Ras responsive element (RRE) very different from the sequence with which we isolated LZ321. We therefore examined the binding of RREB1/LZ321 to different ligands. It bound to the GGTCCT-containing ligand and to the RRE with similar affinities (Kd50-60 nM), but did not bind to a consensus RREB1 binding site. The RREB1/LZ321 protein contains four C2H2zinc-fingers, the C terminal two of which retained specific DNA binding to both ligands. A trimer of the GGTCCT site functioned as an enhancer in both CV-1 and H4IIE-C3 cells. Thus RREB1/LZ321 could function as a downstream activator in the Ras-Raf signaling pathway through different cis -acting elements. A longer human protein, Finb, contains RREB1/LZ321, and there are close homologs in both chicken and Drosophila, arguing that it plays important roles. The ability of transcription factors such as RREB1/LZ321 to bind diverse sequences gives them the potential to regulate previously unsuspected genes. PMID- 10390539 TI - Nucleic acid duplex stability: influence of base composition on cation effects. AB - The effects of counter ion on a nucleic acid duplex stability were investigated. Since a linear free energy relationship for the thermostability of oligonucleotide duplexes between those in 1 M and in 100 mM NaCl-phosphate buffer were observed regardless of whether they are DNA-DNA, RNA-RNA or RNA-DNA duplexes, simple prediction systems for [Delta] G degrees 37as well as T mvalues in 100 mM NaCl-phosphate buffer were established. These predictions were successful with an average error of only 2.4 degrees C for T mand 5. 7% for G degrees 37values. The number of Na+newly bound to a duplex when the duplex forms (-[Delta] n) was significantly influenced by the base composition, and -[Delta] n for d(GCCAGTTAA)/d(TTAACTGGC) was different for MgCl2, CaCl2, BaCl2and MnCl2(from 0.70 to 0.76 with the same order of the duplex stability). Almost no additive effects on the duplex stability was observed for NaCl and MgCl2, suggesting a competitive binding for these cations. The sequence-dependent manner of [Delta] n suggests the presence of preferential base pairs or nearest-neighbor base pairs for the cation binding, which would affect nearest-neighbor parameters. PMID- 10390540 TI - Packaging of DNA by shell crosslinked nanoparticles. AB - We demonstrate compaction of DNA with nanoscale biomimetic constructs which are robust synthetic analogs of globular proteins. These constructs are approximately 15 nm in diameter, shell crosslinked knedel-like (SCKs) nanoparticles, which are prepared by covalent stabilization of amphiphilic di-block co-polymer micelles, self-assembled in an aqueous solution. This synthetic approach yields size controlled nanoparticles of persistent shape and containing positively charged functional groups at and near the particle surface. Such properties allow SCKs to bind with DNA through electrostatic interactions and facilitate reduction of the DNA hydrodynamic diameter through reversible compaction. Compaction of DNA by SCKs was evident in dynamic light scattering experiments and was directly observed by in situ atomic force microscopy. Moreover, enzymatic digestion of the DNA plasmid (pBR322, 4361 bp) by Eco RI was inhibited at low SCK:DNA ratios and prevented when [le]60 DNA bp were bound per SCK. Digestion by Msp I in the presence of SCKs resulted in longer DNA fragments, indicating that not all enzyme cleavage sites were accessible within the DNA/SCK aggregates. These results have implications for the development of vehicles for successful gene therapy applications. PMID- 10390541 TI - SR protein-specific kinase 1 is highly expressed in testis and phosphorylates protamine 1. AB - Arginine/serine protein kinases constitute a novel class of enzymes that can modify arginine/serine (RS) dipeptide motifs. SR splicing factors that are essential for pre-mRNA splicing are among the best characterized proteins that contain RS domains. TwoSRprotein-specifickinases, SRPK1 and SRPK2, have been considered as highly specific for the phosphorylation of these proteins, thereby contributing to splicing regulation. However, despite the fact that SR proteins are more or less conserved among metazoa and have a rather ubiquitous tissue distribution we now demonstrate that SRPK1 is predominantly expressed in testis. In situ expression analysis on transverse sections of adult mouse testis shows that SRPK1 mRNA is abundant in all germinal cells but not in mature spermatozoa. RS kinase activity was found primarily in the cytosol and only minimal activity was detected in the nucleus. In a search for testis-specific substrates of SRPK1 we found that the enzyme phosphorylates human protamine 1 as well as a cytoplasmic pool of SR proteins present in the testis. Protamine 1 belongs to a family of small basic arginine-rich proteins that replace histones during the development of mature spermatozoa. The result of this progressive replacement is the formation of a highly compact chromatin structure devoid of any transcriptional activity. These findings indicate that SRPK1 may have a role not only in pre-mRNA splicing, but also in the condensation of sperm chromatin. PMID- 10390542 TI - Computer analysis of transcription regulatory patterns in completely sequenced bacterial genomes. AB - Recognition of transcription regulation sites (operators) is a hard problem in computational molecular biology. In most cases, small sample size and low degree of sequence conservation preclude the construction of reliable recognition rules. We suggest an approach to this problem based on simultaneous analysis of several related genomes. It appears that as long as a gene coding for a transcription regulator is conserved in the compared bacterial genomes, the regulation of the respective group of genes (regulons) also tends to be maintained. Thus a gene can be confidently predicted to belong to a particular regulon in case not only itself, but also its orthologs in other genomes have candidate operators in the regulatory regions. This provides for a greater sensitivity of operator identification as even relatively weak signals are likely to be functionally relevant when conserved. We use this approach to analyze the purine (PurR), arginine (ArgR) and aromatic amino acid (TrpR and TyrR) regulons of Escherichia coli and Haemophilus influenzae. Candidate binding sites in regulatory regions of the respective H.influenzae genes are identified, a new family of purine transport proteins predicted to belong to the PurR regulon is described, and probable regulation of arginine transport by ArgR is demonstrated. Differences in the regulation of some orthologous genes in E.coli and H.influenzae, in particular the apparent lack of the autoregulation of the purine repressor gene in H.influenzae, are demonstrated. PMID- 10390546 TI - Predominance of Vgamma9/Vdelta2 T lymphocytes in the cerebrospinal fluid of children with tuberculous meningitis: reversal after chemotherapy. AB - BACKGROUND: We analyzed the gammadelta T cell composition and responses in the peripheral blood and cerebrospinal fluid (CSF) of children affected by tuberculous meningitis (TBM) and in control children. MATERIALS AND METHODS: Peripheral blood and CSF samples were stimulated with different phosphoantigens and IL-2, and expansion of Vgamma9/Vdelta2 T cells assessed by FACS analysis. Vgamma9/Vdelta2 lines were obtained by culturing CSF or peripheral blood mononuclear cells (PBMC) in vitro with phosphoantigens and IL-2 for 2 months, and tested for proliferation and cytokine production in response to phosphoantigens. Vdelta2(D)Jdelta junctional sequence length was assessed by PCR. RESULTS: The repertoire of gammadelta T cells from the CSF of TBM patients was characterized by the predominance of Vgamma9/Vdelta2 T lymphocytes, which accounted for >80% of gammadelta T cells. Vgamma9/Vdelta2 cells from the CSF of TBM children responded to different synthetic and natural (mycobacterial) phosphoantigens and produced discrete amounts of IFN-gamma and TNF-alpha. The in vitro expansion of Vgamma9/Vdelta2 T cells from CSF and peripheral blood of TBM patients prominently decreased following chemotherapy, and similarly, the proportion of ex vivo unstimulated Vgamma9/Vdelta2 T cells in CSF of TBM patients decreased to levels detected in the CSF of control subjects. Vdelta2 CDR3 TCR analysis showed that the remaining Vdelta2 cells in the CSF of TBM patients were still polyclonal. CONCLUSIONS: These findings are consistent with an involvement of Vgamma9/Vdelta2 T cells in TBM. http://link. springer ny.com/link/service/journals/00020/bibs/5n5p301. html PMID- 10390547 TI - A strategy to identify genes associated with circulating solid tumor cell survival in peripheral blood. AB - Efforts in metastasis research have centered on the phenotypic and genetic differences between primary site and metastatic site tumors. However, genes that may be used as molecular markers of metastasis in circulating tumor cells remain unidentified. Genes regulating the dissemination and survival of solid tumor cells in the blood, as well as their adaptation to new environments, could be candidates for unique metastatic tumor markers. Differential display (DD) was conducted to compare the blood of tumor-free individuals with the blood of patients with lung, breast, and colon cancers. Twenty-one up-expressed genes in the tumor patient blood samples but none in the tumor-free donor blood samples were identified. Nine of these samples were isolated, amplified, and directly sequenced. A gene AB-1 homologous to a Bcl-2 family member, which might function as an apoptosis inhibitor, was identified. The overexpression of an apoptosis inhibitor in blood from patients with metastatic tumors might be correlated with the capability of solid tumor cells to survive in peripheral blood. This is the first demonstration of the usefulness of comparing control and patient blood samples by DD to find novel potential genetic markers identifying metastasis in the blood. http://link.springer-ny. com/link/service/journals/00020/bibs/5n5p313.html PMID- 10390545 TI - Route and method of delivery of DNA vaccine influence immune responses in mice and non-human primates. AB - BACKGROUND: In spite of the large number of studies that have evaluated DNA-based immunization, few have directly compared the immune responses generated by different routes of immunization, particularly in non-human primates. Here we examine the ability of a hepatitis B surface antigen (HBsAg)-encoding plasmid to induce immune responses in mice and non-human primates (rhesus monkeys: Macaca mulatta) after delivery by a number of routes. MATERIALS AND METHODS: Eight different injected [intraperitoneal (IP), intradermal (ID), intravenous (IV), intramuscular (IM), intraperineal (IPER), subcutaneous (SC), sublingual (SL), vaginal wall (VW)] and six noninjected [intranasal inhalation (INH), intranasal instillation (INS), intrarectal (IR), intravaginal (IVAG), ocular (Oc), oral feeding (oral)] routes and the gene gun (GG) were used to deliver HBsAg expressing plasmid DNA to BALB/c mice. Sera were assessed for HBsAg-specific antibodies (anti-HBs, IgG, IgG1, IgG2a) and cytotoxic T lymphocyte (CTL) activity measured. Three of the most commonly used routes (IM, ID, GG) were compared in rhesus monkeys, also using HBsAg-expressing vectors. Monkeys were immunized with short (0-, 4- and 8-week) or long (0-, 12- and 24-week) intervals between boosts, and in the case of GG, also with different doses, and their sera were assessed for anti-HBs. RESULTS: In one study, anti-HBs were detected in plasma of mice treated by five of eight of the injected and none of the six noninjected routes. The highest levels of anti-HBs were induced by IM and IV injections, although significant titers were also obtained with SL and ID. Each of these routes also induced CTL, as did IPER and VW and one noninjected route (INH) that failed to induce antibodies. In a second study, GG (1.6 microg) was compared to ID and IM (100 microg) delivery. Significant titers were obtained by all routes after only one boost, with the highest levels detected by IM. Delivery to the skin by GG induced exclusively IgG1 antibodies (Th2-like) at 4 weeks and only very low IgG2a levels at later times; ID-immunized mice had predominantly IgG1 at 4 weeks and this changed to mixed IgG1/IgG2a over time. Responses with IM injection (in the leg or tongue) were predominantly IgG2a (Th1-like) at all times. IV injection gave mixed IgG1/IgG2a responses. In monkeys, in the first experiment, 1 mg DNA IM or ID at 0, 4, and 8 weeks gave equivalent anti-HB titers and 0.4 microg at the same times by GG induced lower titers. In the second experiment, 1 mg DNA IM or ID, or 3.2 microg by GG, at 0, 12, and 24 weeks, gave anti-HB values in the hierarchy of GG > IM > ID. Furthermore, high titers were retained after a single immunization in mice but fell off over time in the monkeys, even after boost. CONCLUSIONS: Route of administration of plasmid DNA vaccines influences the strength and nature of immune responses in mice and non-human primates. However, the results in mice were not always predictive of those in monkeys and this is likely true for humans as well. Optimal dose and immunization schedule will most likely vary between species. It is not clear whether results in non-human primates will be predictive of results in humans, thus additional studies are required. http://link.springer-ny.com/link/service/journals/00020/bibs /5n5p287. html PMID- 10390543 TI - BHV-1: new molecular approaches to control a common and widespread infection. AB - BACKGROUND: Herpesviruses are widespread viruses, causing severe infections in both humans and animals. Eradication of herpesviruses is extremely difficult because of their ability to establish latent and life-long infections. However, latency is only one tool that has evolved in herpesviruses to successfully infect their hosts; such viruses display a wide (and still incompletely known) panoply of genes and proteins that are able to counteract immune responses of their hosts. Envelope glycoproteins and cytokine inhibitors are two examples of such weapons. All of these factors make it difficult to develop diagnostics and vaccines, unless they are based on molecular techniques. MATERIALS AND METHODS: Animal herpesviruses, because of their striking similarity to human ones, are suitable models to study the molecular biology of herpesviruses and develop strategies aimed at designing neurotropic live vectors for gene therapy as well as engineered attenuated vaccines. RESULTS: BHV-1 is a neurotropic herpesvirus causing infectious rhinotracheitis (IBR) in cattle. It is a major plague in zootechnics and commercial trade, because of its ability to spread through asymptomatic carrier animals, frozen semen, and embryos. Such portals of infections are also important for human herpesviruses, which mainly cause systemic, eye, and genital tract infections, leading even to the development of cancer. CONCLUSIONS: This review covers both the genetics and molecular biology of BHV-1 and its related herpesviruses. Epidemiology and diagnostic approaches to herpesvirus infections are presented. The role of herpesviruses in gene therapy and a broad introduction to classic and engineered vaccines against herpesviruses are also provided. http://link.springer-ny. com/link/service/journals/00020/bibs/5n5p261.html PMID- 10390551 TI - Abdominal and pelvic needle aspiration biopsies: can we perform them well when using small needles? AB - BACKGROUND: The aim of this study was to compare abdominal fine-needle aspirations (FNAs) performed with large (>/=20-gauge) or small ( 65 but < 75 years, >/= 75 years; CPB time 120 but < 180 minutes, >/= 180 minutes. Both age >/= 75 years (p = 0.024; OR 3.3) and CPB time >/= 180 minutes (p = 0.002; OR 4.2), were found to be predictors of postoperative neurologic damage. Finally, a probability table of stroke risk was obtained with the logistic regression coefficients. A lower stroke probability (0.7%) was calculated in the absence of risk variables and a higher one in the presence of all of them (83.3%). Between these extremes, a total of 158 combinations of stroke probabilities were obtained. We concluded that previous CVA, vasculopathy, emergency operation, and age > 75 years are variously associated with a high risk of nonembolic stroke after myocardial revascularization. A duration of CPB longer than 3 hours strongly increases the probability of neurologic damage in the presence of the aforementioned variables. PMID- 10390581 TI - Gastrointestinal disease following heart transplantation. AB - With advances in heart transplantation, a growing number of recipients are at risk of developing gastrointestinal disease. We reviewed our experience with gastrointestinal disease in 92 patients undergoing 93 heart transplants. All had follow-up, with the median time 4.8 years (range 0.5-9.6 years). During the period of the study we progressively adopted a policy of low immunosuppression aiming toward monotherapy with cyclosporine. Nineteen patients (20.6%) developed 28 diseases related to the gastrointestinal tract. Thirteen patients required 18 surgical interventions, five as emergencies: closure of a duodenal ulcer, five cholecystectomies (one with biliary tract drainage), a sigmoid resection for a diverticulitis with a colovesical fistula, a colostomy followed by a colostomy takedown for an iatrogenic colon perforation, appendectomy, two anorectal procedures, and six abdominal wall herniorrhaphies. At the onset of gastrointestinal disease, 8 patients were on standard triple-drug immunosuppression, all of them within 6 months of transplantation; 13 were on double-drug immunosuppression; and 7 were on cyclosporine alone. All the patients with perforations/fistulas were on steroids. Among the 11 infectious or potentially infectious diseases, 10 were on triple- or double-drug immunosuppression. One death, a patient who was on triple-drug immunosuppression, had a postmortem diagnosis of necrotic and hemorrhagic pancreatitis. Except for an incisional hernia following a laparoscopic cholecystectomy, there was no morbidity and, importantly, no septic complications. We concluded that a low immunosuppression policy is likely to be responsible for the low morbidity and mortality of posttransplant gastrointestinal disease, with a lower incidence of viscous perforation/fistula and infectious gastrointestinal disease. PMID- 10390583 TI - Classification of lymph node metastases from carcinoma of the stomach: comparison of the old (1987) and new (1997) TNM systems. AB - The pN classification of gastric cancer is currently based on the distance of metastatic nodes from the primary tumor (TNM-1987). The UICC (Union Internationale Contre le Cancer) has recently proposed a new classification system based on the number of the involved nodes (TNM-1997). The present prospective study is aimed at verifying whether the two classifications (1) assign approximately a similar rank to individual patients and (2) give comparable prognostic information. The Cox regression model was used to evaluate the prognostic significance of either the distance or the number of positive nodes, controlling for sex, age, site, histology and depth of tumor invasion, in a group of 175 patients who underwent curative surgery for gastric cancer from March 1988 to October 1997. Among the patients classified as N1 and N2 according to TNM-1987, 81.8% (36/44) and 35.8% (19/53), respectively, were coded as N1 and N2 by the new classification. The survival probabilities of N1 and N2 categories were similar in both classifications. The N2 category of TNM-1987 comprised also 10 cases with >15 positive nodes (N3 category of TNM-1997), who presented a large excess mortality (RR = 35.14 with respect to N0). When the site and number of positive nodes are combined in a new variable, both appear to be important from a prognostic point of view. Both anatomic location and number of nodes with metastasis are important predictors of survival in gastric cancer patients. Caution should be used when replacing the old classification with the new one, as they group patients in a different way. PMID- 10390584 TI - Primary retroperitoneal soft tissue sarcomas: results of aggressive surgical treatment. AB - A retrospective study was undertaken to evaluate the results of surgical treatment in a series of patients with primary retroperitoneal sarcomas consecutively treated by the same surgical team. The hospital records of 42 patients with primary retroperitoneal sarcomas who underwent surgical exploration at our unit from 1984 to 1995 were reviewed. A univariate analysis was used to identify the main clinical, pathologic, and treatment-related factors affecting long-term survival. Twenty-five patients (59.6%) underwent radical surgery. The 5 year survival and 5-year disease-free survival after radical resection were 48.1% and 38.8%, respectively. According to the univariate analysis of survival tumor classification (T), stage and gross surgical margins significantly affected prognosis. The study indicates that even though there are predetermined and unmodifiable tumor-related factors, such as tumor classification (T) and stage, that influence survival in patients with retroperitoneal sarcomas, wide surgical excision offers a concrete chance for long-term survival. PMID- 10390585 TI - Perioperative blood transfusion as a prognostic indicator in patients with hepatocellular carcinoma. AB - We studied the relation of perioperative blood transfusion and the outcomes in 175 patients with hepatocellular carcinoma (HCC) who underwent hepatic resection from 1986 to 1994 in our hospital. Hepatectomy was performed in 23 (13.1%) patients with and 152 (86. 9%) without blood transfusions. The cumulative cancer free survival rates for patients who had received blood transfusion was significantly lower than that for patients who had not received blood transfusions (p = 0.003). Further examinations revealed a significant difference in cancer-free survival rates for stage I-II patients (n = 75) of HCC (p = 0.02) but not for stage III-IV patients (n = 56) (p = 0.06). Cox regression analysis for recurrence revealed that blood transfusion was the most significant prognostic indicator (p = 0.001) for recurrence in stage I-II patients but not in stage III-IV patients (p = 0.99). These results suggest that a perioperative blood transfusion may be a significant prognostic indicator for patients with HCC who had underwent hepatectomy, especially in stage I-II patients of HCC. PMID- 10390586 TI - Preoperative assessment of body fluid disturbances in patients with obstructive jaundice. AB - Postoperative renal dysfunction in obstructive jaundice (OJ) patients has been associated with hypovolemia and depletion of the extracellular water compartment (ECW). The aim of the study was to evaluate the preoperative status of body compartments in OJ patients measured by two methods. In a prospective study 39 OJ patients (11 benign and 28 malignant obstructions) were investigated, with 15 healthy subjects used as a control group (CG). Bioelectrical impedance analysis (BIA) determinations and values derived from anthropometric measurements were used to assess body compartment status. The coefficient of variation of BIA was below 4% in both OJ and CG subjects. No differences were found in intracellular water. However total body water (TBW) and ECW were reduced in OJ patients (50.5 +/- 4.6 vs. 56 +/- 8% body weight, p = 0.05; and 21 +/- 4.5 vs. 23.8 +/- 2.5% body weight, p < 0.05, respectively). There were no differences between benign and malignant obstructions. Seventy four percent of OJ patients had an ECW volume below the mean +/- 2 SD in the CG subjects. Anthropometric and BIA determinations correlated closely for TBW measurements in both CG (r = 0.92, p < 0.001) and OJ patients (r = 0.91, p < 0.001). Bland-Altman analysis also showed that for TBW the BIA was in agreement with anthropometry. In the present study, BIA offered a good correlation with anthropometric determinations and was a reliable method for body fluid disturbances assessment in jaundiced patients. PMID- 10390587 TI - Thoracoscopic splanchnicectomy: pilot evaluation of a simple alternative for chronic pancreatic pain control. AB - Achieving adequate pain control in patients with chronic pancreatitis remains a surgical challenge. The quest for a procedure that retains all of the residual pancreatic tissue in the absence of ductal dilatation remains elusive. This study sought to evaluate the feasibility and efficacy of thoracoscopic splanchnicectomy and attempted to outline the surgical anatomy appropriate to an adequate denervation. Of 17 patients considered suitable for the procedure, 16 had a sucessful outcome, which was statistically significant (p < 0.001). The longest follow-up of 30 months suggests that the procedure may be more enduring than percutaneous procedures. However, the surgical anatomy is not predictable owing to the racemose arrangement of the splanchnic fibers, and a long pleurotomy with transection of all medial fibers is necessary to effect denervation. Thoracoscopic splanchnicectomy may effect immediate pain relief with negligible morbidity and absent mortality. Although the follow-up period is short, the patient with the longest follow-up remains pain-free at 30 months. This procedure warrants scrutiny for its role in long-term pancreatic pain control. PMID- 10390588 TI - Value of intravenous cholangiography prior to laparoscopic cholecystectomy. AB - We performed a retrospective study on 163 patients for evaluation of the benefit of intravenous cholangiography prior to laparoscopic cholecystectomy. Radiographic evaluation of the various areas of the biliary system was classified regarding resolution of anatomic structures: well detailed (excellent), impaired image but reliable interpretation possible (good), insufficient contrast with limited assessment (poor), no reliable judgment possible (insufficient). The common bile duct could be described as "good" in 96.3%, whereas the cystic duct could be described as "good" in only 54.6%. Concrements of the gallbladder were recognized in 72.4%, and common bile duct stones were diagnosed in only two of three patients. A distal junction of the cystic duct was found in nine cases, but there was no influence on the following operative procedure. Only one of two patients with a short cystic duct was identified. We found no improvement after routine use of intravenous cholangiography concerning the evidence of common bile duct stones or the avoidance of lesions of the common bile duct. Hence routine use of intravenous cholangiography prior to laparoscopic cholecystectomy is not justified. PMID- 10390589 TI - Laparoscopic common bile duct exploration. AB - Since the introduction of laparoscopic cholecystectomy, the management of common bile duct (CBD) stones has undergone significant change. Preoperative endoscopic retrograde cholangiopancreatography (ERCP) with sphincterotomy is now routinely done in cases where the diagnosis of choledocholithiasis is suspected preoperatively, with clearance of the bile ducts before laparoscopic cholecystectomy. Intraoperative discovery of CBD stones by cholangiography represents a challenge to the surgeon, who must make a decision about when to perform laparoscopic CBD exploration, convert to open surgery, or send the patient for ERCP during the postoperative period. Because ERCP has a definite failure rate, laparoscopic CBD exploration can be a treatment option. Among 2500 laparoscopic cholecystectomies done by our group from January 1991 to June 1997, 50 patients (2%) underwent laparoscopic CBD exploration, 13 by the transcystic technique and 37 by choledocotomy, with a conversion rate of 8% and a hospital stay of 4.3 days. One patient died from complicated pancreatitis following ERCP and unsuccessful extraction of a CBD stone. We obtained our goal of a CBD free of stones in 92% of the cases. We conclude that laparoscopic CBD exploration is an effective method for treating choledocolithiasis that allows management of this pathology in one stage, although it requires advanced laparoscopic skills and adequate equipment. PMID- 10390590 TI - Laparoscopic visualization of the cystic artery anatomy. AB - Uncontrolled bleeding from the cystic artery and its branches is a serious problem that may increase the risk of intraoperative lesions to vital vascular and biliary structures. On laparoscopic visualization anatomic relations are seen differently than during conventional surgery, so proper knowledge of the hepatobiliary triangle anatomic structures under the conditions of laparoscopic visualization is required. We present an original classification of the anatomic variations of the cystic artery into two main groups based on our experience with 200 laparoscopic cholecystectomies, with due consideration of the known anatomicotopographic relations. Group I designates a cystic artery situated within the hepatobiliary triangle on laparoscopic visualization. This group included three types: (1) normally lying cystic artery, found in 147 (73.5%) patients; (2) most common cystic artery variation, manifesting as its doubling, present in 31 (15.5%) patients; and (3) the cystic artery originating from the aberrant right hepatic artery, observed in 11 (5.5%) patients. Group II designates a cystic artery that could not be found within the hepatobiliary triangle on laparoscopic dissection. This group included two types of variation: (1) cystic artery originating from the gastroduodenal artery, found in nine (4. 5%) patients; and (2) cystic artery originating from the left hepatic artery, recorded in two (1%) patients. PMID- 10390591 TI - Diagnostic imaging of early gallbladder cancer: retrospective study of 53 cases. AB - To diagnose early gallbladder carcinoma is difficult but essential to improve the survival of the patients with this cancer. Fifty-three early gallbladder cancers were macroscopically divided into protruding and flat types. The diagnostic devises [ultrasonography (US), computed tomography (CT), and drip infusion cholangiography (DIC)] were compared for their ability of early detection. The specimens were examined cytologically for diagnosis during operation and the p53 protein was investigated. Thirty-three cases were of the protruding type, eighteen of the flat type, and two unclassified. Carcinoma tended to be missed when gallstones were present. Preoperative diagnosis of the flat type was difficult. Tumor location did not always correlate with the preoperative diagnosis. Of the misdiagnosed cases of the protruding type, half were missed with US and CT and were not visualized clearly by DIC. Among the flat type cancers, only three had no abnormal findings by diagnostic imaging. Cytologic examination was effective, and p53 was expressed only in early carcinoma, not in adenoma or dysplasia. Even in the presence of gallstones or cholecystitis, any abnormal findings should make one suspicious of gallbladder cancer. Cytology and p53 expression may be useful for the intraoperative diagnosis, and a combination of diagnostic methods is important. PMID- 10390592 TI - Inflammatory cecal masses in patients presenting with appendicitis. AB - An unexpected inflammatory cecal mass of uncertain etiology encountered during surgery for presumed appendicitis poses a dilemma to the surgeon when deciding the appropriate operative management. A retrospective study was performed to review the pathology and surgical management of this condition. Among 3224 patients who had emergency surgery for a diagnosis of acute appendicitis between January 1990 and December 1997, a group of 52 patients (1.6%) underwent either ileocecal resection or right hemicolectomy for an inflammatory cecal mass of uncertain etiology. The final pathologic diagnosis was cecal diverticulitis in 26 patients (50%), appendiceal phlegmon or abscess in 21 patients (40%), cecal carcinoma in 3 patients (6%), tuberculosis in 1 patient (2%) and schistosomiasis in another patient (2%). Altogether 34 patients underwent ileocecal resection, and 18 patients underwent right hemicolectomy, including the 3 patients with cecal carcinoma. Ileocecal resection was associated with a shorter mean operative time (144 vs. 201 minutes; p < 0.001), a lower morbidity rate (3% vs. 22%; p = 0.043), and a shortened mean postoperative hospital stay (6.8 vs. 11.2 days; p = 0. 011) than right hemicolectomy. There was no mortality in either group. In conclusion, most inflammatory cecal masses are due to benign pathologies and could be managed safely and sufficiently with ileocecal resection. Careful intraoperative assessment including examination of the resected specimen is essential to exclude malignancy, which would require right hemicolectomy. PMID- 10390593 TI - Sigmoidofiberscopic incision plus balloon dilatation for anastomotic cicatricial stricture after anterior resection of the rectum. AB - We describe the procedure and examine the therapeutic efficacy of a combination of sigmoidofiberscopic incision plus balloon dilatation for tubular stricture by thick, long scar tissue at the colorectal anastomosis after anterior resection for rectal cancer. Balloon dilatation alone does not always relieve the strictures, although this method is the usual therapy for this condition. Five patients were identified in whom the stricture was not improved with balloon dilatation alone. Of these five patients, three complained of difficulty defecating, a feeling of incomplete evacuation, residual feces, and lower abdominal fullness. The remaining two patients, who had transverse colostomy to treat major leakage at the anastomosis, showed no symptoms. All five patients underwent the combination therapy described below. Two or three small radial incisions were made in the scar of the stricture with electrocautery under fiberscopic vision. Then the strictural scar was split and loosened bluntly along the incisions over a 15- to 20-minute period with a balloon dilator. This procedure was performed once or twice at a 2-week interval. In all five patients the stricture was improved according to objective criteria. There was also an improvement in the subjective symptoms suffered by three patients. The improvements were maintained over observation periods of 9 to 15 months. No complications were observed. Sigmoidofiberscopic incision plus balloon dilatation is an effective, safe therapy for cicatricial strictures after anterior resection for rectal cancer when the strictures have failed to improve following balloon dilatation alone. PMID- 10390594 TI - Prognostic factors in 2230 Korean colorectal cancer patients: analysis of consecutively operated cases. AB - To define the prognostic factors in Korean colorectal cancer patients, univariate and multivariate analysis were performed on data from 2230 consecutive patients who underwent resection for colorectal cancer at the Seoul National University Hospital. The prognostic variables used for the analysis included patient's age, gender, bowel obstruction, bleeding, symptom duration, preoperative leukocyte count, preoperative serum carcinoembryonic antigen (CEA) level, Dukes' stage, tumor location, tumor size, depth of bowel wall invasion, number of lymph node metastases, histologic differentiation, and gross morphology of tumor. The overall 5-year survival rate was 62%. In the univariate analysis, all the factors except sex, symptom duration, and tumor size were associated with prognosis. Among the factors significant in the univariate analysis, Dukes' stage (p < 0.001), number of lymph node metastasis (p < 0.001), CEA level (p < 0.001), tumor location (p = 0.003), gross morphology of tumor (p = 0.017), and depth of bowel wall invasion (p = 0.031) were significant in the multivariate analysis. Several differences in prognostic factors between colon cancer and rectal cancer were observed. In the multivariate analysis, gross tumor morphology was significant only for colon cancer, and histologic differentiation was significant only for rectal cancer. Lymph node metastasis was an independent prognostic variable for both colon and rectal cancer, but its significance was more prominent for rectal cancer. Although Dukes' stage is the most reliable prognostic predictor, this study shows that other factors (preoperative CEA level, gross morphology of tumor, location of tumor, nodal status) also provide important information for the outcome of the patient. PMID- 10390595 TI - Postoperative changes in thyrotropin-binding inhibitory immunoglobulin level in patients with Graves' disease: is subtotal thyroidectomy a suitable therapeutic option for patients of childbearing age with Graves' disease? AB - Thyroid-stimulating hormone (TSH)-binding inhibitory immunoglobulin (TBII) is thought to be one of the essential causes of Graves' disease, and most cases of neonatal hyperthyroidism can be explained by transplacental passage of TBII. Because surgery is often indicated for patients of childbearing age, it is important to elucidate how surgery reduces TBII levels. Between 1988 and 1991 a total of 946 female patients with Graves' disease underwent subtotal thyroidectomy. Follow-up examination was undertaken at 1, 2, 3, and 4 to 5 years after surgery. At 4 to 5 years after surgery, 76.8% of patients without recurrent overt hyperthyroidism had TBII < 20%. In patients with recurrent hyperthyroidism, TBII increased gradually during follow-up, and they had higher TBII levels than nonrecurrence patients. There were a few euthyroid and hypothyroid patients who had TBII > 60%, and the number of such patients decreased annually. In most of the patients, immunologic remission was obtained by subtotal thyroidectomy except for their having recurrent hyperthyroidism. To acquire immunologic remission, hormonal remission, at least, would be necessary. Because no definite factor other than the size of the thyroid remnant related to postoperative thyroid function was elucidated, near-total thyroidectomy rather than subtotal thyroidectomy is expected to be induced not only hormonal remission but also immunologic remission. It should be noted that a few patients achieved hormonal remission but not immunologic remission. PMID- 10390596 TI - Incidence of malignant melanoma in Auckland, New Zealand: highest rates in the world. AB - The calculation of incidence rates of melanoma in New Zealand has been hampered in the past by incomplete registration of cases. The aim of this study was to document the incidence of melanoma in the Auckland Caucasian population and to define the pathologic characteristics of these lesions. Data were collected for the Auckland region from the New Zealand Cancer Registry and the Auckland Melanoma Unit database for 1995 and combined with census statistics to give the crude and age-standardized rates for invasive melanoma. The results were analyzed by gender, morphology, body site, and thickness. The crude annual incidence for invasive cutaneous malignant melanoma was 77.7/100,000. The age-standardized annual rate was 56.2/100,000 with no statistically significant differences in the rates for males and females. The cumulative risk of developing melanoma over a lifetime, from age 0 to 74, was 5.7% overall. The age-specific rates steadily increase with advancing age. The lesions were generally thin; 64% were less than 0.76 mm, and only 7% were thicker than 3.00 mm. In conclusion, the Caucasian population in the Auckland region has the highest incidence of melanoma in the world. PMID- 10390597 TI - From Billroth to PCV: a century of gastric surgery. AB - The development of gastric surgery is one of the most fascinating chapters in surgical history. The first operations on the stomach were done during the second half of the nineteenth century; at first they were minor procedures but then gradually became more daring, major procedures-albeit with considerable mortality. The work of Theodor Billroth and his pupils ushered in the era of major resectional therapy, first for cancer and later also for ulcer disease. Complications due to the lack of understanding gastric physiology plagued the early days of ulcer surgery, and a variety of modifications tried to remedy these problems. Although the role of the vagus was known through Pavlov's studies, its practical application had to wait until well into the twentieth century. For several decades, resection and vagotomy, separately or combined, were practiced until more sophisticated types of vagotomy began to dominate and replace resection in the surgical treatment of ulcer disease. Resection remained the treatment for cancer. We thus see over a period of 100 years, owing to the increased understanding of physiologic factors, a gradual shift from major resections toward smaller, better directed procedures. The pioneering work of Billroth and his generation, however, must not be forgotten. PMID- 10390598 TI - IMP dehydrogenase : structural aspects of inhibitor binding. AB - Inosine monophosphate dehydrogenase (IMPDH, E.C. 1.1.1.205) is recognized as an important target for both antileukemic and immunosuppressive therapy. IMPDH catalyzes the NAD-dependent oxidation of inosine 5 monophosphate (IMP) to xanthosine 5 monophosphate. Several classes of IMPDH inhibitors are now in use or under development. These include agents that bind at either the substrate site (e.g. ribavirin and mizoribine) or at the NAD site (mycophenolic acid and thiazole-4-carboxamide adenine dinucleotide). All suffer from some degree of toxicity and/or susceptibility to metabolic inactivation. The finding that IMPDH exists as two isoforms, one of which (type II) is induced in tumor cells, has led to the search for potentially more effective isoform-specific agents. Recently, a number of crystal structures of IMPDH have become available. These include structures of the human type II, hamster, Tritrichomonas foetus, Streptococcus pyogenes and Borrelia burgdorferi enzymes. Each structure crystallizes as a tetramer of a/b barrels, with the active site located partly at the monomer monomer interface. The substrate and cofactor bind in a continuous cleft on the C terminal face of each barrel. The IMP base is well positioned to stack against the NAD nicotinamide ring to facilitate hydride transfer. The active site cleft is further bounded by a highly flexible flap and loop. These structures reveal enzyme-ligand interactions which suggest strategies for the design of improved inhibitors. PMID- 10390599 TI - Differential signatures of bacterial and mammalian IMP dehydrogenase enzymes. AB - IMP dehydrogenase (IMPDH) is an essential enzyme of de novo guanine nucleotide synthesis. IMPDH inhibitors have clinical utility as antiviral, anticancer or immunosuppressive agents. The essential nature of this enzyme suggests its therapeutic applications may be extended to the development of antimicrobial agents. Bacterial IMPDH enzymes show biochemical and kinetic characteristics that are different than the mammalian IMPDH enzymes, suggesting IMPDH may be an attractive target for the development of antimicrobial agents. We suggest that the biochemical and kinetic differences between bacterial and mammalian enzymes are a consequence of the variance of specific, identifiable amino acid residues. Identification of these residues or combination of residues that impart this mammalian or bacterial enzyme signature is a prerequisite for the rational identification of agents that specifically target the bacterial enzyme. We used sequence alignments of IMPDH proteins to identify sequence signatures associated with bacterial or eukaryotic IMPDH enzymes. These selections were further refined to discern those likely to have a role in catalysis using information derived from the bacterial and mammalian IMPDH crystal structures and site-specific mutagenesis. Candidate bacterial sequence signatures identified by this process include regions involved in subunit interactions, the active site flap and the NAD binding region. Analysis of sequence alignments in these regions indicates a pattern of catalytic residues conserved in all enzymes and a secondary pattern of amino acid conservation associated with the major phylogenetic groups. Elucidation of the basis for this mammalian/bacterial IMPDH signature will provide insight into the catalytic mechanism of this enzyme and the foundation for the development of highly specific inhibitors. PMID- 10390600 TI - IMP dehydrogenase: mechanism of action and inhibition. AB - Inosine monophosphate dehydrogenase (IMPDH) catalyzes the conversion of IMP to XMP with the concomitant reduction of NAD to NADH. This reaction is the rate limiting step in guanine nucleotide biosynthesis. IMPDH is a proven target for immunosuppressive, anticancer and antiviral chemotherapy, and may also be a target for antimicrobial agents. IMPDH is activated by monovalent cations, and one monovalent cation binding site appears to have been identified. The mechanism of IMPDH involves formation and hydrolysis of a covalent enzyme intermediate (E XMP*) in a reaction reminiscent of glyceraldehyde-3-phosphate dehydrogenase. Substrates bind to IMPDH in a random order, hydride transfer is fast and NADH release precedes hydrolysis of E-XMP*. The hydrolysis of E-XMP* is at least partially rate-limiting. Two inhibitors, mizoribine-monophosphate and a fat base nucleotide appear to act as transition state analogs. In contrast, MPA inhibits by sequestering E-XMP. PMID- 10390601 TI - Consequences of IMP dehydrogenase inhibition, and its relationship to cancer and apoptosis. AB - Inosine 5 -monophosphate dehydrogenase (IMPDH) is a rate-limiting enzyme for the synthesis of GTP and dGTP. Two isoforms of IMPDH have been identified. IMPDH Type I is ubiquitous and predominantly present in normal cells, whereas IMPDH Type II is predominant in malignant cells. IMPDH plays an important role in the expression of cellular genes, such as p53, c-myc and Ki-ras. IMPDH activity is transformation and progression linked in cancer cells. IMPDH inhibitors, tiazofurin, selenazofurin, and benzamide riboside share similar mechanism of action and are metabolized to their respective NAD analogues to exert antitumor activity. Tiazofurin exhibits clinical responses in patients with acute myeloid leukemia and chronic myeloid leukemia in blast crisis. These responses relate to the level of the NAD analogue formed in the leukemic cells. Resistance to tiazofurin and related IMPDH inhibitors relate mainly to a decrease in NMN adenylyltransferase activity. IMPDH inhbitors induce apoptosis. IMPDH inhitors are valuable probes for examining biochemical functions of GTP as they selectively reduce guanylate concentration. Incomplete depletion of cellular GTP level seems to down-regulate G-protein function, thereby inhibit cell growth or induce apoptosis. Inosine 5'-monophosphate dehydrogenase (IMPDH, EC 1.1.1.205) catalyzes the dehydrogenation of IMP to XMP utilizing NAD as the proton acceptor. Studies have demonstrated that IMPDH is a rate-limiting step in the de novo synthesis of guanylates, including GTP and dGTP. The importance of IMPDH is central because dGTP is required for the DNA synthesis and GTP plays a major role not only for the cellular activity but also for cellular regulation. Two isoforms of IMPDH have been demonstrated. IMPDH Type I is ubiquitous and predominately present in normal cells, whereas the IMPDH Type II enzyme is predominant in malignant cells. Although guanylates could be salvaged from guanine by the enzyme hypoxanthine-guanine phosphoribosyltransferase (EC 2.4.2.8), the level of circulating guanine is low in dividing cells and this route is probably insufficient to satisfy the needs of guanylates in the cells. PMID- 10390602 TI - Mizoribine and mycophenolate mofetil. AB - Both mizoribine (MZR) and mycophenolate mofetil (MMF) are immunosuppressive agents that inhibit the proliferation of lymphocytes selectively, via inhibition of IMPDH. MZR is a nucleoside of the imidazole class, isolated from the culture medium of the mold Eupenicillium brefeldianum M-2166. Although this compound has been found to have weak antimicrobial activity against Candida albicans, it has proved ineffective against experimental candidiasis. Unlike azathioprine, this compound is not taken up by nucleic acids in the cell. Instead, after phosphorylation MZR-5 -monophosphate inhibits GMP synthesis by the antagonistic blocking of IMPDH (Ki = 10(-8)M) and GMP- synthetase (Ki =10(-5) M). The drug has been found to inhibit both humoral and cellular immunity, and on this basis it was developed in Japan as an immunosuppressant. MZR has been shown in animal experiments to lack oncogenicity, and has been shown clinically to be associated with a low incidence of severe adverse reactions. MZR has been registered in Japan for the prevention of rejection in renal transplantation, and for the treatment of lupus nephritis, rheumatoid arthritis and the nephrotic syndrome. MMF is the morpholinoethyl ester prodrug of mycophenolic acid (MPA), which was first isolated in 1896 from the culture media of several Penicillium species. MPA has been evaluated for its unique properties as an anticancer, antiviral, antifungal and antibacterial agent, as well as for its therapeutic use in psoriasis and rheumatoid arthritis. MMF was designed to enhance the oral bioavailability of the parent compound. After beneficial effects were observed in animals, the clinical efficacy of MMF as an immunosuppressant in renal transplantation was studied in the United States. In 1995 the US Food and Drug Administration (FDA) approved the use of MMF for the prevention of rejection in renal transplantation, the drug also available on a number of European markets. PMID- 10390603 TI - Nucleoside and non-nucleoside IMP dehydrogenase inhibitors as antitumor and antiviral agents. AB - IMP dehydrogenase (IMPDH) is an enzyme which catalyzes the NAD-dependent conversion of inosine 5 -monophosphate (IMP) to xanthosine 5 -monophosphate (XMP) at the metabolic branch point in the de novo purine nucleotide synthetic pathway. IMPDH was shown to be increased significantly in cancer cells and therefore considered to be a sensitive target for cancer chemotherapy. By blocking the conversion of IMP to XMP, IMPDH inhibitors lead to depletion of the guanylate (GMP, GDP, GTP and dGTP) pools. Two isoforms of human IMPDH, designed type I and type II, have been identified and sequenced. Type I is constitutively expressed and is the predominant isoform in normal cells, while type II is selectively up regulated in neoplastic and replicating cells. Two types of IMPDH inhibitors, endowed with antineoplastic, antiviral and immunosoppressive activity, have been discovered so far: nucleoside inhibitors, such as ribavirin and tiazofurin, and non-nucleoside, such as mycophenolic acid. Ribavirin produces IMPDH inhibition via its anabolite 5 -monophosphate. Tiazofurin inhibits the enzyme after metabolic conversion into thiazole-4-carboxamide adenine dinucleotide (TAD), an analogue of the cofactor NAD. It was hypothesized that the inhibitory activity of tiazofurin is due to an attractive electrostatic interaction between the heterocyclic sulphur atom and the furanose oxygen 1 which constrain rotation about the C-glycosidic bond in tiazofurin and in its active anabolite TAD. To check this hypothesis, we studied several C-nucleosides related to tiazofurin and their NAD analogues. Non-nucleoside IMPDH inhibitors are also reviewed. PMID- 10390604 TI - Mechanism-based design of inosine 5-monophosphate dehydrogenase inhibitors: synthesis and biological activities of 5-ethynyl-1-beta-D-ribofuranosylimidazole 4-carboxamide (EICAR) and its derivatives. AB - Inosine 5 -monophosphate dehydrogenase (IMPDH) catalyzes the nicotinamide adenine dinucleotide (NAD)-dependent oxidation of inosine 5 -monophosphate (IMP) to xanthosine 5 -monophosphate (XMP), and is one of the key rate-determining enzymes of de novo guanine nucleotide biosynthesis in mammalian systems. Based on the proposed catalytic mechanism of IMPDH, we designed and synthesized 5-ethynyl-1 beta-D-ribofuranosylimidazole-4-carboxamide (5b, EICAR) from 5-amino-1-beta-D ribofuranosylimida-zole-4-carboxamide (AICAR) via palladium chemistry. EICAR is a potent cytostatic agent that inhibits various tumor cells in culture including human solid tumor cells in vitro and in vivo. EICAR also showed broad-spectrum antiviral activities, about 10- to 100-fold greater than those of ribavirin. An examination of the structure-activity relationships revealed that an alkynyl group, especially an ethynyl group at the 5-position, is important for its activity due to the inhibition of IMPDH. The mode of action of EICAR is also discussed. PMID- 10390605 TI - Novel mycophenolic adenine bis(phosphonate)s as potential immunosuppressants. AB - Mycophenolic acid (MPA) is the most potent and specific inhibitor of inosine monophosphate dehydrogenase (IMPDH). This compound was reported to bind the NAD site of IMPDH and mimic the binding of nicotinamide moiety of nicotinamide adenine dicnucleotide. We linked MPA derivatives with the adenine moiety of NAD through a methylenebis(phonphonate) birdge to form novel mycophenolic adenine dinucleotides (MADs) which resemble well the intact natural cofactor. The MAD analogues differ by the length of the side chain (linker) between the aromatic ring of mycophenolic derivative and the beta-phosphorus atom of the adenosine bis(phosphonate) moiety. Regardless of the linker size, MADs were found to be potent inhibitors of human IMPDH type I and type II with Ki's = 0.25-0.52 microM, an order of magnitude less potent than MPA itself (Ki = 0.01-0.04 microM). The growth of K562 cells was inhibited by MPA (IC50 = 0.03 microM) and the MAD analogues (IC50 = 0.01-1.15 microM) with a similar potency. Accordingly, a suppression of alloantigen- induced proliferation of human lymphocytes by the MAD analogues at concentration of 10-20 microM was equally effective as that observed for MPA. In contrast to MPA, MAD analogues were found to be resistant to glucuronidation in vitro. Since therapeutic potential of MPA is limited by its undesirable glucuronidation, the glucuronidation- resistant MAD analogues may be superior immunosuppressants if they are not glucuronidated in vivo. PMID- 10390606 TI - Succinimide and saccharin-based enzyme-activated inhibitors of serine proteases. AB - The inhibition of human leukocyte elastase (HLE), and other serine proteases, by succinimide and saccharin-based compounds is reviewed. The succinimide compounds are unique in that the inactivating species is generated within the enzyme active site via a molecular rearrangement. The related saccharin derivatives also inactivate serine proteases by an enzyme-activated mechanism. Those factors effecting the potency, selectivity and stability of these important classes of inhibitor are discussed. PMID- 10390607 TI - Current trends in the development of nitric oxide donors. AB - Nitric oxide (NO) is an important messenger molecule involved in many pathological and physiological processes within the mammalian body. Exogenous NO sources constitute a powerful way to supplement NO when the body can not generate enough for normal biological functions. In this article, general aspects on NO and NO donors are reviewed. Major focus is placed on recent developments of novel NO donors, NO releasing device(s) as well as innovative improvements to current NO donors. Finally, an outlook on future NO donor development is provided. PMID- 10390608 TI - Transferrin As A targeting ligand for liposomes and anticancer drugs. AB - In cancer treatment, one of the approaches is targeting of the drug to tumor cells via receptor specific ligands. Transferrin (molecular weight 80,000) has been used as a ligand for delivering anticancer drugs or drug containing liposomes mostly due to the increased number of transferrin (trf) receptors found on tumor cells as compared to normal cells. Transferrin was linked to methotrexate (MTX) containing small unilamellar liposomes and its activity was compared to antitransferrin receptor antibody (7D-3) linked to MTX liposomes. In each of these conjugates, the method of coupling was the same and a disulphide linkage was formed between the ligand and MTX liposomes. No significant differences in the potency of 7D-3 conjugate or trf conjugate with MTX liposomes were observed in studies performed in vitro against various human tumor cell lines (Hela, KB and Colon). Trf was also linked to adriamycin via a schiff base which was formed by using glutaraldehyde. This conjugate was found to be effective in vitro against various human tumors (Lovo, HL-60, SW 403 and Hep2) and also in vivo against H-mesothelioma tumors. Transferrin receptor has also been used for gene delivery. Gene delivery to K562 haematopoietic leukaemic cells was achieved by using a transferrin-polycation (poly-L-lysine or protamine) conjugate. This review will cover the various important applications of transferrin based drug delivery formulations in the chemotherapy of cancer and the related work performed in our and other laboratories. PMID- 10390609 TI - A new class of diacidic nonpeptide angiotensin II receptor antagonists: candesartan cilexetil. AB - Blockade of the action of angiotensin II (AII) has long been a target for development of novel antihypertensive agents. We recently discovered a novel class of potent nonpeptide AII receptor antagonists, benzimidazole-7-carboxylic acids including candesartan. Candesartan is a highly potent and insurmountable angiotensin II type-1 receptor (AT1)-selective antagonist. Structure-activity relationship (SAR) studies revealed that the adjacent arrangement of a lipophilic substituent, a tetrazolylbiphenylmethyl moiety and a carboxyl group was the important structural requirement for potent AII antagonistic activity. The benzimidazole ring was found to be one of the most suitable templates arranging these three essential components in correct direction. Especially, the presence of a carboxyl group at the 7-position was found to be essential for insurmountable antagonism. Although candesartan is a very potent AII antagonist, it was found to be absorbed rather inefficiently upon oral administration. To improve bioavailability (BA) of candesartan, chemical modification was examined to yield candesartan cilexetil, a prodrug of candesartan. Candesartan cilexetil is a potent and long-acting blocker that provides effective 24 hr blood pressure control. Our alternative research efforts to improve oral BA was performed by replacement of the tetrazole ring in candesartan by other new acidic bioisosteric heterocyclic rings to find the nonprodrug AII antagonist TAK-536, bearing 5-oxo 1,2,4-oxadiazole ring, which was as potent and orally active as candesartan cilexetil. PMID- 10390610 TI - Does picosecond protein dynamics have survival value? PMID- 10390612 TI - The recently reported NIbeta domain is already known as the Calx-beta motif. PMID- 10390611 TI - Vitamin C: poison, prophylactic or panacea? AB - Ascorbate is an essential enzyme cofactor but is often also regarded as an important antioxidant in vivo, protecting against cancer by scavenging DNA damaging reactive oxygen species. Recent studies suggest that ascorbate sometimes increases DNA damage in humans. Although there is no evidence that any of these effects are deleterious to humans, we might need to change our thinking about the mechanisms of the antioxidant action of ascorbate in vivo. PMID- 10390613 TI - Domains in plexins: links to integrins and transcription factors. PMID- 10390614 TI - The FF domain: a novel motif that often accompanies WW domains. PMID- 10390615 TI - Retrograde protein translocation: ERADication of secretory proteins in health and disease. AB - Eukaryotic cells have a complex degradation machinery that eliminates misfolded or unassembled secretory proteins from the endoplasmic reticulum (ER). The proteins are retained in an ER/pre-Golgi compartment and then hydrolysed by the cytosolic ubiquitin-proteasome system. This requires retrograde translocation of proteins from the ER back to the cytoplasm, which is mediated by Sec61, the central component of the ER protein-import channel. This proteolytic pathway prevents a potentially lethal aggregation of secretory proteins; however, several viruses misuse it to escape detection, and bacterial and plant toxins might also exploit it. Underactive or overactive ER degradation machinery contributes to the pathogenesis of several severe human diseases. PMID- 10390616 TI - DNA recombination: the replication connection. AB - Chromosomal double-strand breaks (DSBs) arise after exposure to ionizing radiation or enzymatic cleavage, but especially during the process of DNA replication itself. Homologous recombination plays a critical role in repair of such DSBs. There has been significant progress in our understanding of two processes that occur in DSB repair: gene conversion and recombination-dependent DNA replication. Recent evidence suggests that gene conversion and break-induced replication are related processes that both begin with the establishment of a replication fork in which both leading- and lagging-strand synthesis occur. There has also been much progress in characterization of the biochemical roles of recombination proteins that are highly conserved from yeast to humans. PMID- 10390617 TI - Nucleotide sequence databases: a gold mine for biologists. AB - The rapid expansion of nucleotide sequence data available in public databases is revolutionizing biomedical research. These databases have a variety of uses, including the discovery of novel genes, identification of homologous genes, analysis of alternative splicing, chromosomal localization of genes, and detection of polymorphisms. Data sets such as the human transcript map will undoubtedly accelerate identification of candidate genes in positional-cloning approaches. Careful in silico analysis can significantly reduce the amount of lab work required. Approximately half of all human genes are represented in these databases; therefore, one need not wait for the entire human genome to be sequenced before performing genome-wide studies. PMID- 10390618 TI - Signaling routes to CREM and CREB: plasticity in transcriptional activation. AB - The CREB and CREM transcription factors are activated by phosphorylation of a key serine residue by kinases stimulated by cyclic AMP, Ca2+, growth factors and stress signals. Phosphorylation allows recruitment of CREB-binding protein (CBP), a large co-activator that contacts the general transcriptional machinery. Studies of the physiological roles played by CREB and CREM have uncovered novel routes of transcriptional activation. For example, in male germ cells CREM is not phosphorylated but associates with ACT, a member of the LIM-only class of proteins that has intrinsic transcriptional activity. Thus, in some circumstances, CREM can bypass the classical requirement for phosphorylation and association with CBP. PMID- 10390619 TI - MolSurfer: two-dimensional maps for navigating three-dimensional structures of proteins. PMID- 10390620 TI - David Phillips and the origin of structural enzymology. PMID- 10390621 TI - UFD1L and CDC45L: a role in DiGeorge syndrome and related phenotypes? AB - Molecular genetics is contributing to the understanding of normal and abnormal cardiovascular development and morphogenesis. Deletions of chromosome 22q11.2 have been associated with distinct phenotypes that result from a failure to form derivatives of third and fourth branchial arches, including DiGeorge syndrome (DGS) and velo-cardio-facial syndrome (VCFS). The biochemical mechanisms underlying these phenotypes remain undetermined. A recent study provides new insight into the mechanism by which gene deletions produce the DGS and VCFS phenotypes. PMID- 10390622 TI - Reply: role of the dHAND-UFD1L pathway PMID- 10390623 TI - Inferring the direction of evolutionary changes of genomic base composition. PMID- 10390624 TI - Suppression mechanisms: themes from variations. AB - Suppressor analysis is a commonly used strategy to identify functional relationships between genes that might not have been revealed through other genetic or biochemical means. Many mechanisms that explain the phenomenon of genetic suppression have been described, but the wide variety of possible mechanisms can present a challenge to defining the relationship between a suppressor and the original gene. This article provides a broad framework for classifying suppression mechanisms and describes a series of genetic tests that can be applied to determine the most likely mechanism of suppression. PMID- 10390625 TI - Monogenic traits are not simple: lessons from phenylketonuria. AB - The classification of genetic disease into chromosomal, monogenic and multifactorial categories is an oversimplification. Phenylketonuria (PKU) is a classic 'monogenic' autosomal recessive disease in which mutation at the human PAH locus was deemed sufficient to explain the impaired function of the enzyme phenylalanine hydroxylase (enzymic phenotype), the attendant hyperphenylalaninemia (metabolic phenotype) and the resultant mental retardation (cognitive phenotype). In the era of molecular genetics, expectations for a consistently close correlation between the mutant genotype and variant phenotype have been somewhat disappointed, and PKU is used here to illustrate how and why this might be the case. So-called monogenic traits do, indeed, conform to long accepted ideas about the expression of 'major' loci and their importance in determining parameters of phenotype, but the associated features are as complex, in their own ways, as those in so-called complex traits. PMID- 10390626 TI - Cell fate and organogenesis in bacteria. AB - Intercellular signaling through the Notch receptor and its ligands leads to the spatial differentiation of cell fate in vertebrates and invertebrates. In Myxococcus xanthus, fruiting-body development requires the transmission of a cell bound intercellular signal by the protein called C-factor, which is functionally equivalent to the eukaryotic Notch ligands. Functional parallels between these two signaling systems include strong positive and negative feedback, and a consequent role in spatial differentiation. Consideration of these parallels enables us to make testable experimental predictions about Notch and C-signaling. PMID- 10390627 TI - The evolution of new structures: clues from plant cytoskeletal genes. AB - How large numbers of genes were recruited simultaneously to build new organ structures is one of the greatest puzzles in evolutionary biology. Here, we present data suggesting that the vegetative and reproductive classes of actins and other cytoskeletal proteins arose concurrently with the macroevolutionary divergence of leaves and reproductive structures in the earliest land plants. That the cytoskeleton is essential for physically programming the development of organs and tissues is well established. Thus, we propose that this regulatory dichotomy represents an ancient landmark event in the global regulation of hundreds of higher-plant genes, an event that is linked to the macroevolution of plant vegetative and reproductive organs. The recent availability of sequence and expression data for large numbers of plant genes should make it possible to dissect this and other major macroevolutionary events. PMID- 10390628 TI - Making effective use of human genomic sequence data. PMID- 10390629 TI - Endogenous retroviruses: are they the cause of multiple sclerosis? PMID- 10390630 TI - Prion diseases and the central dogma of molecular biology. PMID- 10390632 TI - Response from McFadden and waller PMID- 10390631 TI - Evolutionary pathway of the apicomplexan plastids and its implications. PMID- 10390633 TI - Cancer (or lack thereof) and viruses. PMID- 10390634 TI - Stress-inducible mechanisms of life-span extension in yeast, eubacteria and metazoans. PMID- 10390636 TI - Response from sokurenko and dykhuizen PMID- 10390635 TI - Divergence in the background: hard to measure but the key to being the best of the best. PMID- 10390637 TI - The cellulosome concept as an efficient microbial strategy for the degradation of insoluble polysaccharides. AB - The cellulosome is an extracellular supramolecular machine that can efficiently degrade crystalline cellulosic substrates and associated plant cell wall polysaccharides. The cellulosome arrangement can also promote adhesion to the insoluble substrate, thus providing individual microbial cells with a direct competitive advantage in the utilization of the soluble hydrolysis products. PMID- 10390638 TI - Variation and evolution of the citric-acid cycle: a genomic perspective. AB - The presence of genes encoding enzymes involved in the citric-acid cycle has been studied in 19 completely sequenced genomes. In the majority of species, the cycle appears to be incomplete or absent. Several distinct, incomplete cycles reflect adaptations to different environments. Their distribution over the phylogenetic tree hints at precursors in the evolution of the citric-acid cycle. PMID- 10390639 TI - The cytolethal distending toxin family. AB - Cytolethal distending toxins are produced by a small but diverse group of bacterial pathogens. This newly discovered toxin family can cause a variety of mammalian cells to become irreversibly blocked in the G2 phase of the cell cycle. How this novel effect is accomplished is unknown but the study of these fascinating toxins promises to reveal new methods of host-pathogen interaction. PMID- 10390640 TI - Evolution of Wolbachia pipientis transmission dynamics in insects. AB - Wolbachia pipientis is an intracellular bacterial parasite of arthropods that enhances its transmission by manipulating host reproduction, most commonly by inducing cytoplasmic incompatibility. The discovery of isolates with modified cytoplasmic incompatibility phenotypes and others with novel virulence properties is an indication of the potential breadth of evolutionary strategies employed by Wolbachia. PMID- 10390642 TI - Now available onlineellipsis trends in pharmacological sciences PMID- 10390641 TI - Corrigendum PMID- 10390643 TI - Recent advances in the pharmacology of quaternary salts of bicuculline. PMID- 10390644 TI - Protease-activated receptors: PAR4 and counting: how long is the course? PMID- 10390645 TI - Ligand efficacy and affinity in an interacting 7TM receptor model. AB - Quantitative understanding of the activation of G protein-coupled receptors is based mostly on some theoretical models that describe the interaction between ligand and protein partners and the activation process of the receptor. All of these models provide different definitions for observable affinity or efficacy. However, the property common to such parameters defined in the context of these models is that they are always independent of the concentration of the receptor molecule. This is based on the assumption that receptors do not interact with each other appreciably. In this article, experimental evidence for which this assumption does not seem to apply is discussed and an oligomerization model for seven-transmembrane-domain receptors that explains the relationship between receptor concentration, apparent affinity and efficacy is provided. PMID- 10390646 TI - Partial agonists and G protein-coupled receptor desensitization. AB - Weak or partial agonists induce less desensitization of G protein-coupled receptors (GPCRs) than do strong agonists. However, there have been few attempts to relate partial agonism quantitatively with the various parameters of agonist induced desensitization, and to elucidate the mechanisms involved. Our understanding of how the treatment of cells and tissues with partial agonists affects their capacity to activate receptors is based on continued progress in defining partial agonism and the mechanisms of desensitization in which protein kinases, phosphatases, endocytosis and recycling play various roles. In this review, current research concerning partial-agonist-induced desensitization of GPCRs and the nature of partial agonism is summarized, and an attempt is made to put the existing knowledge into a working hypothesis concerning the mechanisms that account for the reduced desensitization in response to partial agonists. PMID- 10390647 TI - Pharmacological and biochemical interactions between opioids and cannabinoids. AB - Opioids and cannabinoids are among the most widely consumed drugs of abuse in humans. A number of studies have shown that both types of drugs share several pharmacological properties, including hypothermia, sedation, hypotension, inhibition of both intestinal motility and locomotor activity and, in particular, antinociception. Moreover, phenomena of cross-tolerance or mutual potentiation of some of these pharmacological effects have been reported. In recent years, these phenomena have supported the possible existence of functional links in the mechanisms of action of both types of drugs. The present review addresses the recent advances in the study of pharmacological interactions between opioids and cannabinoids, focusing on two aspects: antinociception and drug addiction. The potential biochemical mechanisms involved in these pharmacological interactions are also discussed together with possible therapeutic implications of opioid cannabinoid interactions. PMID- 10390648 TI - 7TM receptors: the splicing on the cake. AB - Within a given family of seven transmembrane domain (7TM) receptors, functional diversity is most often afforded by the existence of multiple receptor subtypes, each encoded by a distinct gene. However, it is now clear that the existence of introns in genes encoding some members of a receptor family provides scope for additional diversity by virtue of splicing events that result in the formation of different receptor mRNAs and consequently distinct receptor isoforms. A large number of 7TM receptor splice variants have now been shown to exist. In this article, the current data on alternatively spliced variants for hormone and neurotransmitter 7TMs are reviewed, their potential physiological importance considered and some of the issues pertaining to the classification and nomenclature of receptor isoforms produced in this way are addressed. PMID- 10390649 TI - Neurotensin and neurotensin receptors. AB - Neurotensin is a brain and gastrointestinal peptide that fulfils many central and peripheral functions through its interaction with specific receptors. Three subtypes of neurotensin receptors have been cloned. Two of them belong to the family of G protein-coupled receptors, whereas the third one is an entirely new type of neuropeptide receptor and is identical to gp95/sortilin, a 100 kDa protein with a single transmembrane domain. In this review, the present knowledge regarding the molecular and pharmacological properties of the three cloned neurotensin receptors is summarized and the relationship between these receptors and the known pharmacological effects of neurotensin is discussed. PMID- 10390650 TI - Response from schulz and moore PMID- 10390651 TI - Guidelines for the psychotherapies in comprehensive psychiatric care: a discussion paper. Working Group 2 of the Canadian Psychiatric Association Psychotherapies Steering Committee. PMID- 10390652 TI - The practice and roles of the psychotherapies: a discussion paper. Working Group 1 of the Canadian Psychiatric Association Psychotherapies Steering Committee. PMID- 10390653 TI - Hepatitis C: implications for the general physician. AB - Approximately 4 million people in the United States are affected with chronic hepatitis C--20%-25% of whom will develop cirrhosis. Determination of the stage of disease and extent of cirrhosis requires liver biopsy. In cirrhotic patients, regular screening for hepatocellular carcinoma is indicated. PMID- 10390654 TI - Inaccuracies in patient medical histories. AB - Of patients who completed the same medical history questionnaire twice within a certain time period, 66% had at least 1 significant omission in their history. Consideration is given to the merits of a national medical history computerized database. PMID- 10390655 TI - Sexually transmitted diseases in adolescents: an update. AB - Sexually transmitted diseases are a significant cause of morbidity for U.S. adolescents. Recent advances in diagnosis may lead to enhanced recognition of infected individuals, particularly those who lack symptoms. New approaches to therapy have simplified treatment and may enhance compliance. PMID- 10390656 TI - Pulmonary complications of cancer therapy. AB - The development of pulmonary disease as a result of cancer therapy is an increasingly recognized clinical problem. Chemotherapeutic drugs can induce an acute pneumonitis, pulmonary edema, and pulmonary fibrosis, as well as a variety of other pulmonary diseases in cancer patients. PMID- 10390657 TI - Chronic neck syndromes: an update. AB - Chronic neck pain syndromes are recognized as myofascial disorders in the main. Often such syndromes follow trauma, and there are significant psychological and secondary gain issues that impact on the outcome of therapy. Comparison with cervical radicular syndromes is discussed. PMID- 10390658 TI - Mind-body therapy in the management and prevention of coronary disease. AB - Conventional mind-body therapy has been proven a valuable noninvasive way to manage coronary disease. Yoga practice, especially, has been found to be valuable in preventing adverse outcomes of coronary disease by improving resistance to stress. PMID- 10390659 TI - Two studies: long-term cognitive decline after bypass surgery; Alzheimer's gene linked to earlier bypass. PMID- 10390660 TI - Chirality and the mode of action of anaesthetics. PMID- 10390661 TI - Sevoflurane requirement for laparoscopic tubal ligation: an electroencephalographic bispectral study. AB - It has been shown in healthy volunteers that a concentration of volatile anaesthetic lower than 1 minimum alveolar concentration provides unconsciousness. We tested the hypothesis that, using the electroencephalogram bispectral index, less than 1 minimum alveolar concentration of sevoflurane can produce unconsciousness in patients. Anaesthesia was induced and maintained with sevoflurane in N2O and O2 (33%) in 32 ASA I-II women undergoing laparoscopic tubal ligation. For the first patient, the sevoflurane concentration was adjusted to 1 minimum alveolar concentration with an end-tidal concentration of 0.7%. The electroencephalogram bispectral index values were used to determine the concentration to be used for the next patient. The ED50 (effective dose) measured using end tidal concentrations of sevoflurane for laparoscopic tubal ligation in a 40-year-old patient was 0.70% (CI 95%: 0.63-0.77) and ED95 0.83% (CI 95%: 0.75 0.90). None of the patients had any operation-associated recall. It is concluded that the sevoflurane concentration needed for laparoscopic tubal ligation is not lower than 1 minimum alveolar concentration. PMID- 10390662 TI - Dimenhydrinate for prevention of post-operative nausea and vomiting in female in patients. AB - Dimenhydrinate is an inexpensive antihistaminic drug, that is frequently used as an anti-emetic during anaesthesia. The popularity of the drug is contrasted by the lack of modern studies concerning its efficacy in reducing the incidence of post-operative nausea and vomiting. Thus, dimenhydrinate was compared with placebo in this prospective, randomized, double-blind study. One hundred and thirty-three female in-patients were studied. They were stratified according to the type of surgery (laparoscopic cholecystectomy, thyroid resection or knee arthroscopy) to ensure an homogeneous distribution in both groups. General anaesthesia was induced with etomidate, fentanyl, vecuronium and maintained with enflurane in N2O/O2. Neuromuscular block was reversed with pyridostigmine/atropine. Patients in the dimenhydrinate group (n = 67) received 62 mg dimenhydrinate intravenously after induction of anaesthesia. Placebo patients (n = 66) received saline. Administration of dimenhydrinate (and placebo) was repeated three times during the 48-h study to mitigate the short half-life of the drug. Post-operative analgesia and anti-emetic rescue medication was standardized. Episodes of vomiting, retching and the need for additional anti emetics were recorded. Nausea was assessed using a 10-cm visual analogue scale. Post-operative nausea and vomiting was rated as 'none', 'mild', 'moderate' and 'severe' using a fixed scoring algorithm. There were no differences between the two groups with regard to biometric data, type of surgery and distribution of risk factors for developing post-operative nausea and vomiting. In the dimenhydrinate group, more patients remained completely free from post-operative nausea and vomiting compared with placebo (dimenhydrinate: 38.8%; placebo: 15.1%; P = 0.004). The incidence of severe post-operative nausea and vomiting was also reduced from 39.4% to 14.9%. No relevant side effects were observed. Intra operative dimenhydrinate, followed by three further administrations after surgery, reduces the incidence and the severity of post-operative nausea and vomiting without side effects. However, there still remained an unacceptable high number of patients who were not prevented completely from experiencing post operative nausea and vomiting. PMID- 10390663 TI - Droperidol-supplemented anaesthesia decreases post-operative nausea and vomiting but impairs post-operative mood and well-being. AB - Post-operative nausea and vomiting is distressing for patients and can cause dissatisfaction and impaired well-being in the post-operative period. This study examined the question whether the reduced incidence of post-operative nausea and vomiting inevitably translates into improved clinical status and well-being. In this context high doses of droperidol were investigated. On the one hand, droperidol is known to be a powerful anti-emetic, but on the other hand there is concern about psychological effects, both in the pre- and the post-operative period. In this prospective randomized double-blinded study, droperidol (5-7.5 mg) was compared with midazolam (5-7.5 mg) used to supplement fentanyl-N2O based anaesthesia, with respect to post-operative mood and well-being using a psychological questionnaire (Bf-S-test). Furthermore, the incidence of post operative nausea and vomiting was recorded. Out of 160 patients undergoing thyroidectomy and laparoscopic cholecystectomy, data from 150 patients were analysed. The administration of droperidol significantly lowered the incidence of post-operative nausea and vomiting from 77.8% to 55.1% compared with midazolam (P = 0.0059; chi 2-test). Although post-operative nausea and vomiting is an independent risk factor for post-operative discomfort and bad mood, patients receiving droperidol showed impaired well-being 6 h after surgery. Well-being scores returned to pre-operative base-line values and did not differ between the two groups 24 and 48 h post-operatively. The reduced incidence of post-operative nausea and vomiting achieved with high dose droperidol does not equate with increased post-operative well-being. It is an important point at issue to decide whether smaller doses of droperidol that are commonly used for anti-emetic therapy are free of these side effects. PMID- 10390664 TI - Seven-year review of requests for epidural blood patches for headache after dural puncture: referral patterns and the effectiveness of blood patches. AB - A review was undertaken of all 190 patients who were referred over 7 years, from 1991 to 1997 inclusive, for an epidural blood patch as a treatment for headache after dural puncture. The patterns of referral and symptoms, the distributions of age and gender and the effectiveness of the blood patch were examined. Most of the referrals (n = 153) were after deliberate diagnostic dural puncture in neurology and neuroradiology, with a minority (n = 28) used for anaesthesia and obstetrics, which were mostly inadvertent. Another nine cases were related to placement of an intrathecal catheter. The numbers of referrals per year reached a maximum in 1995 before falling again, a curious inverse relation to the number of invasive neuro-radiological diagnostic procedures. Most of the patients were between 30 and 50-years-old, with 25 younger than 30 and 14 older than 60. Women accounted for 70% of the referrals for headache, although the gender ratio amongst patients subjected to at risk procedures appeared closer to 50:50. Neckache accompanied the headache in 85% of cases, auditory problems were volunteered by three patients and one patient had diplopia for 6 weeks. Of the 190 patients who were referred, 186 received at least one patch, the symptoms in the remaining four being too mild or atypical to warrant blood patch treatment. This provided initial relief in all but two patients, one of whom received a further epidural blood patch with no effect. There was sustained relief of symptoms in 136 and a partial relapse in 38 patients, which resolved without needing any further blood patch. A second patch was provided for seven patients and a third for three patients, of whom two were cured. Of the patients who needed more than one blood patch, nine were after inadvertent dural puncture with a Tuohy needle and, of these patients, six were in labour. A total of 200 patches were provided in all for the 186 patients and all but three patients had a satisfactory outcome. Epidural blood patches are effective in treating headache after dural puncture, but less successful than is commonly believed, especially after inadvertent dural taps. A relapse after treatment does not always require a second patch. Specialities other than anaesthesia seemed reluctant to accept the benefits in both cost and comfort of using needles of improved design for dural puncture. PMID- 10390665 TI - Anti-emetic efficacy of prophylactic granisetron compared with perphenazine for the prevention of post-operative vomiting in children. AB - We have compared the efficacy of granisetron with perphenazine in the prevention of vomiting after tonsillectomy with or without adenoidectomy in children. In a prospective, randomized, double-blind study, 90 paediatric patients, ASA I, aged 4-10 years, received granisetron 40 mg kg-1 or perphenazine 70 mg kg-1 (n = 45 each) intravenously immediately after an inhalation induction of anaesthesia. A standard general anaesthetic technique was employed throughout. A complete response, defined as no emesis with no need for another rescue antiemetic, during the first 3 h (0-3 h) after anesthesia was 87% with granisetron and 78% with perphenazine (P = 0.204). The corresponding incidence during the next 21 h (3-24 h) after anaesthesia was 87% and 62% (P = 0.007). No clinically serious adverse events were observed in any of the groups. We conclude that granisetron is a better anti-emetic than perphenazine for the long-term prevention of post operative vomiting in children undergoing general anaesthesia for tonsillectomy. PMID- 10390666 TI - Practice of epidural analgesia for labour pain: a German survey. AB - Epidural analgesia is one of the preferred methods of analgesia for labour. The aim of the present survey was to evaluate current practice in obstetric analgesia in departments of anaesthesia and to make a comparison with former surveys from Germany and other countries. Questionnaires on the practice of pain relief, especially epidural analgesia, during labour and delivery were sent to 1178 anaesthetic departments in Germany in the second half of 1996. Five hundred and thirty-two completed replies were received, which represent 46.9% of all German obstetric units. The majority of the departments of anaesthesia practising epidural analgesia have an epidural rate of less than 10% and 10.2% of the departments do not offer this method to their parturients. In 86.8% of all units performing epidural analgesia, the epidural catheter is placed by an anaesthetist. Only 6.5% of the units provide a 24-h epidural service which is exclusively assigned to labour and delivery. In 77.8% of the units, this service is not exclusively assigned to obstetrics, but also to other duties. Of the obstetric units offering epidural analgesia, 14.7% have no epidural service at night. Plain local anaesthetics for epidural analgesia are used by 55.9% of the departments, a combination of local anaesthetics with epidural opioids by 28.7%. Epidural analgesia is predominantly (82.2%) maintained by intermittent bolus administration. Although the rate of epidural analgesia increased during recent decades, this method is not offered to all parturients. Further improvements in the use of epidural analgesia for labour seem to be necessary. PMID- 10390667 TI - The anaesthetic potency of propofol in the rat is reduced by simultaneous intravenous administration of lignocaine. AB - Lignocaine added to the anaesthetic preparation Diprivan reduces propofol induced pain on injection. This effect is due to a drop in pH which decreases the content of propofol in the aqueous phase of the soya bean emulsion. This in turn changes the electrostatic forces in the emulsion and destabilization occurs. The effect of lignocaine on the anaesthetic potency of propofol was validated in a randomized blind study in the rat. The induction dose of 1% propofol mixed with 1% lignocaine (10 + 1) was significantly higher when compared with the induction dose of propofol 1% given after a separate injection of 1% lignocaine (9.4 +/- 5.5 vs. 5.6 +/- 5.2 mg; P < 0.05). The duration of sleep was shorter in rats injected with propofol 1% mixed with lignocaine 1% (10 + 1) compared with those given 1% lignocaine and 1% propofol in separate injections (160 +/- 181 vs. 375 +/- 202 s; P < 0.05). The anaesthetic potency of propofol was not significantly changed by the addition of either saline or hydrochloric acid. The anaesthesia inducing effect was not time-dependent. A similar lower potency was observed for a solution stored for 4 h compared with one freshly prepared, although sleeping time was longer (9.2 +/- 6.8 mg; 428 +/- 110 s) as compared with the 4 h mixture. The results indicate that lignocaine altered the propofol preparation. The reduced anaesthetic potency of propofol after addition of lignocaine is not due to the resultant drop in pH, which is known to occur. PMID- 10390668 TI - Effects of graded suppression of the EEG with propofol on the neurological outcome following incomplete cerebral ischaemia in rats. AB - We evaluated the relation between dose and response for the neuroprotective effect of propofol in a rat model with incomplete cerebral ischaemia. For clarification of the mechanism of neuroprotection, plasma catecholamines and tumour necrosis factor-alpha levels were measured. Three doses (low, moderate and high-dose) of propofol were tested. These produced, respectively, a low amplitude, slowing and a burst-suppression pattern of electroencephalographic activity. Incomplete cerebral ischaemia was produced by right carotid artery occlusion combined with haemorrhagic hypotension (35 mmHg) for 30 min. Neurological outcome at 72 h post-ischaemia in the high-dose group was significantly better than that in both low-dose and moderate-dose groups. Propofol exhibited a trend in the dose-related attenuation of the increases in plasma adrenaline and noradrenaline during ischaemia. Tumour necrosis factor alpha increased during and after ischaemia in all groups with no intergroup differences. The results indicate that a burst-suppression dose of propofol provides neuroprotection. The protective effect can not be completely explained by the attenuating effect on circulating catecholamines. PMID- 10390670 TI - Anaesthesia for Sturge-Weber syndrome. AB - A 6-month-old boy with Sturge-Weber syndrome was scheduled for congenital glaucoma and left buphthalmus surgery. Physical examination revealed haemangioma throughout the right trigeminal nerve, congenital glaucoma, left megalocornea and bilateral buphthalmus. Examination of the eye was performed under general anaesthesia, was followed 2 days later by trabeculotomy. No premedication was given to the patient. After induction of anaesthesia with halothane, O2 and N2O muscle relaxation was achieved with atracurium and he was intubated gently. No difference was observed in vital signs during surgery. At the end of the operation he was given oxygen 100% and extubated, muscle relaxant reversal was with atropine and neostigmine. No complication was observed in the post-operative period. PMID- 10390669 TI - Spinal anaesthesia with gamma hydroxybutyrate. A study in a rat model. AB - Gamma hydroxybutyric acid, a central inhibitory neurotransmitter and a cerebral metabolite of gamma-aminobutyric acid, is present in high concentrations in the mammalian hypothalamus and basal ganglia. Its sodium salt gamma hydroxybutyrate has been effectively used as an intravenous anaesthetic agent, and as an oral sedative, and in the management of the alcohol withdrawal syndrome. In an animal model, using 72 Wistar strain rats allocated to one of six groups of 12 animals each, with implanted lumbar intrathecal catheters, we examined whether gamma hydroxybutyrate, 20% 40 microL (32 mg kg-1) administered alone or combined with fentanyl, gamma hydroxybutyrate 20% 20 microL (16 mg kg-1), fentanyl 0.005% 20 microL (4 mg kg-1) as an intrathecal bolus, provides intraoperative anaesthesia, comparable with that produced by intrathecal lignocaine. We demonstrated that gamma hydroxybutyrate, given by an intrathecal bolus in the rat model, produced reversible segmental antinociception, together with muscular relaxation of the abdominal wall and rear limbs. This is accompanied by moderate sedation without haemodynamic or respiratory depression. This agent may thus be promising for use as a spinal anaesthetic drug. PMID- 10390671 TI - Paradoxical air embolism during orthoptic liver transplantation: diagnosis by transoesophageal echocardiography. AB - We describe a case of paradoxical air embolism during orthotopic liver transplantation, early diagnosis, using intra-operative transoesophageal echocardiography after a circulatory failure, allowed early management by hyperbaric oxygen therapy. PMID- 10390672 TI - Botulism with respiratory insufficiency requiring extra corporeal carbon dioxide removal. AB - Despite a low incidence of botulism in the industrialized world some cases occasionally occur in Germany after eating contaminated food. Because botulism is rarely seen, most physicians are unfamiliar with its early recognition and treatment. However, immediate intensive care treatment is important. We report the case of a previously healthy 54-year-old female who developed signs of botulism after eating vacuum packed smoked fish and developed severe respiratory insufficiency with difficult carbon dioxide elimination in the days following. PMID- 10390673 TI - Fatal massive intra-operative pulmonary embolism while placing a patient in the surgical position. PMID- 10390674 TI - Empirical medical ethics. PMID- 10390675 TI - Reply to Farsides's editorial: palliative care--a euthanasia-free zone. PMID- 10390676 TI - Equitable rationing of highly specialised health care services for children: a perspective from South Africa. AB - The principles of equality and equity, respectively in the Bill of Rights and the white paper on health, provide the moral and legal foundations for future health care for children in South Africa. However, given extreme health care need and scarce resources, the government faces formidable obstacles if it hopes to achieve a just allocation of public health care resources, especially among children in need of highly specialised health care. In this regard, there is a dearth of moral analysis which is practically useful in the South African situation. We offer a set of moral considerations to guide the macro-allocation of highly specialised public health care services among South Africa's children. We also mention moral considerations which should inform micro-allocation. PMID- 10390677 TI - Genetically determined obesity in Prader-Willi syndrome: the ethics and legality of treatment. AB - A central characteristic of people with Prader-Willi Syndrome (PWS) is an apparent insatiable appetite leading to severe overeating and the potential for marked obesity and associated serious health problems and premature death. This behaviour may be due to the effects of the genetic defect resulting from the chromosome 15 abnormalities associated with the syndrome. We examine the ethical and legal dilemmas that can arise in the care of people with PWS. A tension exists between a genetic deterministic perspective and that of individual choice. We conclude that the determination of the capacity of a person with PWS to make decisions about his/her eating behaviour and to control that behaviour is of particular importance in resolving this dilemma. If the person is found to lack capacity, the common law principles of acting in a person's "best interests" using the "least restrictive alternative" may be helpful. Allowing serious weight gain in the absence of careful consideration of these issues is an abdication of responsibility. PMID- 10390678 TI - Sterilisation of incompetent mentally handicapped persons: a model for decision making. AB - Doctors are regularly confronted with requests for sterilisation of mentally handicapped people who cannot give consent for themselves. They ought to act in a medical vacuum because there doesn't exist a consensus about a model for decision making on this matter. In this article a model for decision making is proposed, based on a review of the literature and our own research data. We have attempted to select and classify certain factors which could enable us to arrive at an ethically justifiable method of making a medical decision. In doing so we distinguish two major criteria: heredity and parenting competence, and six minor criteria: conception risk, IQ, age, personality, medical aspects and prognosis and finally support and guidance for the mentally handicapped person. The major criteria give rise to a "situation of necessity". In this situation the physician is confronted with a conflict of values and interests. The minor criteria are of an entirely different ethical order. They can only be considered once the major criteria have created a "situation of necessity". Ultimately it comes down to deciding whether the benefits of sterilisation outweigh the drawbacks and whether the means are appropriate to the end, where efficient contraception is the end and irreversible sterilisation is the means. PMID- 10390679 TI - A problem for the idea of voluntary euthanasia. AB - I question whether, in those cases where physician-assisted suicide is invoked to alleviate unbearable pain and suffering, there can be such a thing as voluntary euthanasia. The problem is that when a patient asks to die under such conditions there is good reason to think that the decision to die is compelled by the pain, and hence not freely chosen. Since the choice to die was not made freely it is inadvisable for physicians to act in accordance with it, for this may be contrary to the patient's genuine wishes. Thus, what were thought to be cases of voluntary euthanasia might actually be instances of involuntary euthanasia. PMID- 10390680 TI - Advance directives are the solution to Dr Campbell's problem for voluntary euthanasia. AB - Dr Neil Campbell suggests that when patients suffering extremes of protracted pain ask for help to end their lives, their requests should be discounted as made under compulsion. I contend that the doctors concerned should be referred to and then act upon advance directives made by those patients when of sound and calm mind and afflicted by no such intolerable compulsion. PMID- 10390681 TI - Protective truthfulness: the Chinese way of safeguarding patients in informed treatment decisions. AB - The first part of this paper examines the practice of informed treatment decisions in the protective medical system in China today. The second part examines how health care professionals in China perceive and carry out their responsibilities when relaying information to vulnerable patients, based on the findings of an empirical study that I had undertaken to examine the moral experience of nurses in practice situations. In the Chinese medical ethics tradition, refinement [jing] in skills and sincerity [cheng] in relating to patients are two cardinal virtues that health care professionals are required to possess. This notion of absolute sincerity carries a strong sense of parental protectiveness. The empirical findings reveal that most nurses are ambivalent about telling the truth to patients. Truth-telling would become an insincere act if a patient were to lose hope and confidence in life after learning of his or her disease. In this system of protective medical care, it is arguable as to whose interests are being protected: the patient, the family or the hospital. I would suggest that the interests of the hospital and the family members who legitimately represent the patient's interests are being honoured, but at the expense of the patient's right to know. PMID- 10390682 TI - Abandonment of terminally ill patients in the Byzantine era. An ancient tradition? AB - Our research on the texts of the Byzantine historians and chroniclers revealed an apparently curious phenomenon, namely, the abandonment of terminally ill emperors by their physicians when the latter realised that they could not offer any further treatment. This attitude tallies with the mentality of the ancient Greek physicians, who even in Hippocratic times thought the treatment and care of the terminally ill to be a challenge to nature and hubris to the gods. Nevertheless, it is a very curious attitude in the light of the concepts of the Christian Byzantine physicians who, according to the doctrines of the Christian religion, should have been imbued with the spirit of philanthropy and love for their fellowmen. The meticulous analysis of three examples of abandonment of Byzantine emperors, and especially that of Alexius I Comnenus, by their physicians reveals that this custom, following ancient pagan ethics, in those times took on a ritualised form without any significant or real content. PMID- 10390683 TI - Research, ethics and conflicts of interest. AB - In this paper, I have tried to develop a critique of committee procedures and conflict of interest within research advisory committees and ethical review committees (ERCs). There are specific features of conflict of interest in medical research. Scientists, communities and the subjects of research all have legitimate stakeholdings. The interests of medical scientists are particularly complex, since they are justified by the moral and physical welfare of their research subjects, while the reputations and incomes of scientists depend on the success of their science. Tensions of this kind must at times produce conflict of interest. It is important to recognise that conflicts of interest may unwittingly lead to manipulation of research subjects and their lay representatives on research committees. It is equally important to recognise distinctions between the legal and moral aspects of conflict of interest. Some practical suggestions are made which may go some way towards resolving these difficulties. They indicate what might be needed to ensure the validity of ethical discourse, and to reduce the risks associated with conflict of interest. PMID- 10390684 TI - Can the written information to research subjects be improved?--an empirical study. AB - OBJECTIVES: To study whether linguistic analysis and changes in information leaflets can improve readability and understanding. DESIGN: Randomised, controlled study. Two information leaflets concerned with trials of drugs for conditions/diseases which are commonly known were modified, and the original was tested against the revised version. SETTING: Denmark. PARTICIPANTS: 235 persons in the relevant age groups. MAIN MEASURES: Readability and understanding of contents. RESULTS: Both readability and understanding of contents was improved: readability with regard to both information leaflets and understanding with regard to one of the leaflets. CONCLUSION: The results show that both readability and understanding can be improved by increased attention to the linguistic features of the information. PMID- 10390685 TI - Performance of research ethics committees in Spain. A prospective study of 100 applications for clinical trial protocols on medicines. AB - OBJECTIVES: To review the characteristics and performance of research ethics committees in Spain in the evaluation of multicentre clinical trial drug protocols. DESIGN: A prospective study of 100 applications. SETTING: Forty-one committees reviewing clinical trial protocols, involving 50 hospitals in 25 cities. MAIN MEASURES: Protocol-related features, characteristics of research ethics committees and evaluation dynamics. RESULTS: The 100 applications involved 15 protocols (of which 12 were multinational) with 12 drugs. Committees met monthly (except one). They had a mean number of 12 members, requested a mean of six complete dossiers and nine additional copies of the protocol with a mean deadline of 14 days before the meeting. All applications were approved except three (two of the three were open-label long-term safety trials rejected by the same committee), which were approved by the other committees involved. The mean time from submission to approval was 64 days. The mean time from submission to arrival of the approval document at our offices was 85 days. Twenty-five committees raised queries for 38 of the 97 finally approved applications. Impact of evaluation fee, number of members, queries raised and experience of committees on timings were not statistically significant. CONCLUSION: Obtaining ethical approval is time-consuming. There is much diversity in the research ethics committees' performance. A remarkable delay (> 20 days) exists between the decision and the arrival of the written approval, suggesting administrative or organisational problems. PMID- 10390687 TI - Teaching literature and medicine. PMID- 10390686 TI - Responses by four Local Research Ethics Committees to submitted proposals. AB - BACKGROUND: There is relatively little research concerning the processes whereby Local Research Ethics Committees discharge their responsibilities towards society, potential participants and investigators. OBJECTIVES: To examine the criteria used by LRECs in arriving at their decisions concerning approval of research protocols through an analysis of letters sent to investigators. DESIGN: Four LRECs each provided copies of 50 letters sent to investigators after their submitted proposals had been considered by the committees. These letters were subjected to a content analysis, in which specific comments and requests for additional information and changes in the protocols were recorded and compared. FINDINGS: Overall 24% of proposals were approved without request for changes or clarifications, but this varied by committee: one committee approved only 6% of proposals without change or clarification while the others ranged from 26% to 32%. The content analyses of responses indicated that they could be placed into four categories: (i) further information for the committee to aid in their deliberations; (ii) requests for changes to the design or justification for the design used; (iii) changes to the information sheets provided to potential participants; and (iv) changes to consent procedures. Of these, alterations to information sheets were the most common type of request. These four types of response could be seen as safeguarding the wellbeing of potential participants (the principle of non-maleficence), of promoting the scientific validity of the research (the principle of beneficence), and of enhancing the rights of potential participants (the principle of autonomy). CONCLUSIONS: The committees were consistent in the types of requests they made of investigators, which can be seen as attempts to protect participants' rights and ensure the scientific validity of studies. Without an analysis of the proposals sent to the committees, however, it is difficult to account for the variation in the requirements set by the committees before approval was given. PMID- 10390688 TI - Workplace influenza immunization update. PMID- 10390689 TI - Helicobacter pylori infection in a large company. PMID- 10390690 TI - Genotoxicity from domestic use of organophosphate pesticides. PMID- 10390691 TI - Aspartame and symptoms of carpal tunnel syndrome. PMID- 10390692 TI - Vibration, pathophysiology, and industrial control. PMID- 10390693 TI - Health effects of the 1991 Kuwait oil fires: a survey of US army troops. AB - The burning of oil wells in Kuwait in 1991 discharged a high volume of potentially toxic pollutants into the air. To determine whether there were health related complaints associated with having lived and worked there, questionnaires were administered to 1599 soldiers after their return from a 3-month mission in Kuwait. Symptoms occurring before, during, and after the mission were queried. Compared with baseline, symptoms reported more frequently for the Kuwait period were eye and upper respiratory tract irritation, shortness of breath, cough, rashes, and fatigue. Symptoms were associated with reported proximity to oil fires, and their incidence generally decreased after the soldiers left Kuwait. Oil-fire smoke is one of several possible factors that may have contributed to the reporting of symptoms. PMID- 10390694 TI - Emerging research on the treatment of Gulf War veterans' illnesses. AB - Much of what is known about Gulf War veterans' illnesses is a result of the investment in research by the federal government. Since 1993, the Research Working Group of the Federal Interagency Persian Gulf Veterans Coordinating Board has guided the federal research program. Based on this research, Hodgson and Kipin conclude that the symptom-based conditions reported by Gulf War veterans could be treated through the use of a technique called cognitive behavioral therapy. This past year, the Department of Veterans Affairs launched the largest multisite, randomized, controlled treatment trial of the effectiveness of exercise and cognitive behavioral therapy in relieving the symptoms of ill Gulf War veterans. Despite this important step forward, the Department and its Research Working Group partners continue to explore all aspects of Gulf War veterans' illnesses. PMID- 10390695 TI - Gulf War illnesses: causation and treatment. AB - Soldiers returning from the Gulf War in 1991 described a range of symptoms, including some consistent with the chronic fatigue syndrome, fibromyalgia, and multiple chemical sensitivity. Well-defined adverse health events attributable to service in the Gulf occurred. However, controlled epidemiological studies in Gulf War veterans and controls describe significant excesses of symptoms that were not clearly associated with pathologic disease. At least 12% of veterans currently receive some form of disability from the Department of Veterans Affairs. A number of reports outline theories proposed to explain the excess, but few are scientifically supported. Management guidelines for this spectrum of disorders resembles that of many of "emerging overlap syndromes," including multiple chemical sensitivity, chronic fatigue syndrome, and fibromyalgia. They include the establishment of a trusting doctor-patient relationship, negotiations around a common ground of scientific and etiologic beliefs, non-labeling of the disorder, and work toward recovery in the absence of clear etiologic answers. PMID- 10390696 TI - A thermal physiological comparison of two HAZMAT protective ensembles with and without active convective cooling. AB - Wearing impermeable garments for hazardous materials (HAZMAT) cleanup can often present a health and safety problem for the wearer. Even short duration cleanup activities can produce heat-stress injuries in HAZMAT workers. It was hypothesized that an internal cooling system might increase worker productivity and decrease the likelihood of heat-stress injuries in typical HAZMAT operations. Two HAZMAT protective ensembles were compared during subjects' treadmill exercise. The different ensembles were created using two different suits: a Trelleborg vapor protective suit representative of current HAZMAT suits and a prototype suit developed by engineers at the National Aeronautics and Space Administration (NASA). The two life-support systems used were a current technology Interspiro Spirolite breathing apparatus and a liquid air breathing system that also provided convective cooling. Twelve local members of a HAZMAT team served as test subjects. They were fully instrumented to allow a complete physiological comparison of their theramal responses to the different ensembles. Results showed that cooling from the liquid air system significantly decreased thermal stress. The results of the subjective evaluations of new design features in the prototype suit were also highly favorable. Incorporation of these new design features could lead to significant operational advantages in the future. PMID- 10390697 TI - A geographical information system technique for record-matching in a study of cancer deaths in welders. AB - Retrospective studies are frequently complicated by incomplete worker identifiers. We encountered this problem when evaluating the risk of cancer death in a welding cohort. We dealt with it by developing birth-date ranges for each welder with unknown birth dates and using geographical information system techniques in conjunction with last name, gender, and birth-date range to assign death certificates to welders on the basis of residential proximity to the worksite. Deaths for total malignant cancers and lung/tracheobronchial/pleural cancers among these welders were not significantly different from those in county, state, and US populations, using standardized mortality ratios. The ratios in our study subjects were consistent with ratios found in other published welder cohorts. PMID- 10390698 TI - Impact of a modern firefighting protective uniform on the incidence and severity of burn injuries in New York City firefighters. AB - The New York City Fire Department (FDNY) is the largest fire department in the United States, with over 11,000 firefighters. In 1994, FDNY changed to a modern firefighting protective uniform. The major difference between traditional and modern uniforms is that modern uniforms include both protective over-coat and over-pant, whereas traditional uniforms include only the over-coat. Furthermore, modern uniforms are manufactured using improved thermal protective textiles that meet or exceed current National Fire Protection Association standards for structural firefighting. The purpose of this study was to determine the impact of the modern uniform on the incidence and severity of FDNY burn injuries. We also evaluated the incidence and severity of other non-burn injuries to determine whether there was serious adverse impact. The number of lower-extremity burns decreased by 85% when 2 years' experience while wearing the modern uniform was compared with 2 years while wearing the traditional uniform. Upper-extremity burns and head burns decreased by 65% and 40%, respectively. Severity indicators (days lost to medical leave, hospital admissions, and skin grafts) for lower- and upper-extremity burn injuries were all substantially reduced. This occurred without significant change in the incidence or severity of trunk burns, heat exhaustion, inhalation injuries (actually decreased), or cardiac events. The reduction in the incidence and severity of burn injuries, the major occupational injury affecting this workforce, has been so dramatic and without untoward effects that the introduction of the modern uniform must be characterized as a sentinel event in the history of firefighter health and safety. PMID- 10390699 TI - Accelerated silicosis with mixed-dust pneumoconiosis in a hard-metal grinder. AB - We describe a fatal case of accelerated silicosis with a component of mixed-dust pneumoconiosis in a young hard-metal grinder that we believe is the first case of its kind in Israel and one of the rare cases reported worldwide. The patient's diagnosis was based on typical features: restrictive lung function, abnormal chest roentgenogram suggesting lung fibrosis, a history of exposure to silica and hard metals, bronchoalveolar lavage (BAL) fluid findings, and mineralogical studies. BAL cells showed an abundance of giant multinucleated macrophages. The CD4/CD8 ratio of T lymphocytes was 1.1, with a high percentage of CD8 and CD8/38 positive cells (37% suppressor/cytotoxic and 12% cytotoxic T lymphocytes, respectively). mRNA transcripts isolated from BAL cells were positive for interleukin-1 (IL-1) and transforming growth factor (TGF) Il-5, IL-2, and IL-10 but not for IL-6, IL-4, and interferon. Polarizing light microscopic studies of BAL and induced sputum cells showed polarizing particles, which are typical for silica. Mineralogical studies of electron microscopy performed on BAL fluid and on dust collected at the patient's workstation revealed silica particles as well as aluminum-titanium and other particles. The latter might have contributed to the patient's lung disease. PMID- 10390700 TI - Spontaneous abortions among Finnish flight attendants. AB - We conducted a retrospective cohort study to investigate whether work as a cabin attendant is related to an increased risk for spontaneous abortion. Data on female cabin crew members were linked to medical records on pregnancies. There were 1751 eligible pregnancies for the final analysis. Flight attendants who worked during early pregnancy had a slightly elevated risk of spontaneous abortion, as compared with attendants who were pregnant outside a time span of active flying (odds ratio [OR] = 1.3; 95% confidence interval [CI], 0.9 to 1.8). During the earliest years of the study period (1973 through 1977), the risk seemed to be decreased (OR = 0.4; 95% CI, 0.2 to 1.1), whereas during the later years (1978 through 1994) the risk was increased (OR = 1.6; 95% CI, 1.1 to 2.4). The results are in agreement with earlier studies, showing suggestive evidence of a slightly increased risk of spontaneous abortion among female cabin crew members. PMID- 10390701 TI - Working conditions and health effects of ethylene oxide exposure at hospital sterilization sites. AB - Ethylene oxide (EtO) is a powerful disinfectant and sterilant for heat-sensitive surgical items and instruments. Its use in hospitals constitutes an important source of occupational exposure that is sometimes underestimated, such as in cases of EtO device malfunction when the safety rules of procedure are not strictly followed or when individual or collective protective equipment is lacking. We carried out a descriptive study of the health care workers who were assigned to EtO sterilization units of the Lille University Hospital Centre in Lille, France (n = 16). Before the modification of the sterilization units in the development of a single, central sterilization site, we studied the workplaces, occupational conditions, and work procedures of the health care workers exposed to EtO. The aim was to assess the risk of EtO overexposure of the workers in order to improve workers' health and security in the future sterilization center. The study was based on a physical examination, a questionnaire covering each subject's personal and occupational history, and a complete ocular examination. For occupational conditions, the studies of each workplace were also performed by the occupational physician. Area and personal breathing air samplings were performed at each exposure site. Fourteen of the 16 operators had posterior and anterior subcapsular lens opacities, three of which seemed to be directly and primarily related to occupational exposure; the other ten seemed to be rather common and compatible with age. High levels of EtO exposure were reported in the oldest site (90 parts per million [ppm] during the changing of the gas bottle), where exposure often exceeded French threshold limits (permissible exposure limit: 1 ppm 8-hour time-weighted average (TWA) in air; short-term excursion limit: 5 ppm 15-minute TWA in air), or the current US recommended and legal exposure limits for EtO advocated by the Occupational Safety and Health Administration and the American Conference of Governmental Industrial Hygienists (permissible exposure limit: 1 ppm 8-hour TWA in air; excursion limit: 5 ppm 15 minute TWA in air), and the National Institute for Occupational Safety and Health standard (recommended exposure limits: 0.1 ppm 8-hour TWA in air; 5 ppm 10-minute TWA in air). The faults in the work processes, such as interruption of the sterilization cycle and disregard for the use of protective devices, were very common. PMID- 10390702 TI - Absence of polyneuropathy among workers previously diagnosed with solvent-induced toxic encephalopathy. AB - An association between polyneuropathy and occupational exposure to trichloroethylene, trichloroethane, perchloroethylene, or similar solvents alone or in combination is controversial. We sought to determine whether workers previously diagnosed with solvent-induced toxic encephalopathy had objective evidence of polyneuropathy. Thirty railroad workers previously diagnosed with toxic encephalopathy were examined in the context of litigation against their employers. All described long-term occupational solvent exposure averaging 20 years in duration (range, 10 to 29 years) and producing acute intoxication on a regular basis. The diagnosis of subclinical or clinical polyneuropathy was established using a combination of symptoms, signs, and nerve conduction study (NCS) measures, consistent with standard clinical practice. Potential confounders were identified. NCS results were compared with historical controls, including unexposed workers matched by gender, age, and body mass index. Dose-response relationships were evaluated using simple linear and stepwise regression models. Three workers fulfilled clinical polyneuropathy criteria. The only worker fulfilling NCS criteria for confirmed clinical polyneuropathy had diabetes mellitus. Mean NCS values for most measures were similar to control values, and existing differences in sensory amplitudes disappeared when compared with the matched control group. NCS measures were not significantly influenced by exposure duration or job title. Separation into groups on the basis of the presence or absence of polyneuropathy symptoms, previous diagnosis of polyneuropathy, disability status, and severity or type of encephalopathy did not demonstrate significant NCS differences. The complaints of these workers claiming neurotoxic injury from occupational solvent exposure are not explained by peripheral nervous system dysfunction. PMID- 10390704 TI - A state trauma registry as a tool for occupational injury surveillance. AB - The goal of occupational surveillance is to identify and determine the magnitude of work-related disease and injury and workplace hazards for the purposes of focusing prevention programs and tracking their effectiveness. There are a number of databases that collect information on pieces of the puzzle of workplace exposure and adverse health outcomes. Other than that for the fatalities, none of these datasets specifically describes the most severe occupational injuries or their attendant disability. The goal of this study was to evaluate the usefulness of the Illinois Trauma Registry (ITR) in the surveillance of occupational injuries. The entire dataset of the ITR was obtained from the Illinois Department of Public Health for the years 1993 and 1994. The occupational injuries were extracted and frequency distributions were determined for all demographic and health variables. Background population, employment, and death-rate data were obtained for the purpose of rate calculation and for comparison of raw data. Mean costs for acute occupational injuries were calculated. There were 5844 occupational cases, comprising 6.7% of the total group. The majority of injuries had occurred in males (86%), in urban settings (81%), and were of the "blunt" injury type. External cause (coded according to the International Classification of Diseases, 9th Revision, External Injury) categories for work-related injuries were "Cut/Struck," 39%; "Falls," 36%; "Transportation," 12%; "Environmental," 6%; "Violence," 3%; and the remainder, 5%. By definition, all cases were admitted to the hospital, with 62% classified as "minor," 28% "moderate," and the remaining 11% "severe" to "life threatening." Surgery was performed in 54%, and admission to a monitored bed or the intensive care unit occurred in 15%. Although 93% were discharged home, only 54% ambulated independently. Seven percent were not independent with regard to self-feeding status. The mean hospital charge was $10,802 (standard deviation, $31,438). A pyramidal model of the place of ITR cases in the universe of occupational injuries is presented. The ITR contains a unique set of variables that broaden our understanding of serious work-related injuries. It is recommended that these variables--"occupation," "type of industry," and "nature of injury"--be added to the ITR so that it may be linked with other databases to check its validity and completeness and to enhance its value in occupational surveillance. PMID- 10390703 TI - Concentration of marijuana metabolites in the urine after ingestion of hemp seed tea. AB - To determine whether ingestion of hemp seed tea could result in positive urine drug screens for cannabinoids, volunteers were recruited to donate urine after consuming hemp seed or placebo tea. Among the 22 participants, 10 ingested 12 ounces of hemp seed tea, 10 ingested 24 ounces, and 2 ingested 12 ounces of placebo tea. Urine cannabinoid specimens were obtained at baseline and at 4, 8, and 24 hours after ingestion. A total of 10 specimens had trace quantities of cannabinoids detected in 7 subjects on gas chromatography/mass spectrometry testing, all below the Department of Transportation cutoff level of 15 ng/mL. These results demonstrate that under the conditions of this study, hemp seed tea consumption can result in detectable urine cannabinoids but would not trigger a positive EMIT or gas chromatography/mass spectrometry urine drug test for cannabinoids. PMID- 10390705 TI - Plasma p53 protein and anti-p53 antibody expression in vinyl chloride monomer workers in Taiwan. AB - Vinyl chloride (VC) workers are known to be at risk for development of angiosarcoma of the liver (ASL), a rare tumor. Previously, a study of p53 gene mutations in tumors of VC-exposed workers found that 50% of liver angiosarcomas contained such mutations. Mutant p53 oncoprotein and anti-p53 antibodies can also be found in the sera of ASL patients and VC-exposed workers without cancer. Workers in Taiwan have also been exposed to VC, and some have contracted liver tumors. In this study, we used enzyme-linked immunosorbent assays to detect mutant p53 protein and anti-p53 antibodies in the plasma of VC-exposed workers in Taiwan. Thirty-three of 251 (13.2%) VC-workers tested positive for the p53 overexpression (10% with positive mutant p53 protein and 3.6% with positive anti p53) in their plasma, but only 2 of 36 controls (5.6%) tested positive (2.8% with positive mutant p53 protein and 2.8% with positive anti-p53). There was a significant association between cumulative VC exposure concentration and positive p53 expression (P = 0.032) among VC workers after we adjusted for age, hepatitis, drinking, and smoking status. In summary, P53 overexpression (mutant p53 protein or anti-p53 antibody) can be found in the plasma of VC workers in Taiwan, and a significant dose-response relationship exists between plasma p53 overexpression and VC cumulative exposure concentration. PMID- 10390706 TI - Practice not limited to dental public health. PMID- 10390707 TI - Fluoride varnish: a useful new tool for public health dentistry. AB - Since first shown to be effective by Bibby in 1942, professionally applied topical fluorides have been used successfully as a caries-preventive intervention. In the United States, the acidulated phosphate fluoride (APF) gels have been the most widely used agent for over two decades. While effective, APF has several practical disadvantages, including unpleasant taste and the risk of fluoride overingestion. The use of APF on infants and very young children is not practical or safe. Recently, fluoride varnish has become available in the United States. It has been used widely in Europe and Scandinavia for 25 years. Its effectiveness and safety are documented in over 50 clinical trials. Fluoride varnish is easy to apply, is well accepted by children, and eliminates the risk of overingestion of fluoride. Fluoride varnish is approved by the FDA as a "device" and must be used "off label" for the prevention of caries. Because of the large body of published data documenting its effectiveness and safety, there is no legal risk in using fluoride varnish off label. In fact, the American Dental Association has granted its seal of approval to Duraphat, one of the varnish products. Varnish offers considerable advantages in the dental public health setting. Of particular note, it is practical and safe to apply to the teeth of infants and very young children. PMID- 10390708 TI - Changes in dental health and dental health habits from 3 to 5 years of age. AB - OBJECTIVES: This study sought to determine how dental health and dental health habits change from 3 to 5 years of age and to consider whether preventive dental health care helped in preventing or halting caries in children. METHODS: The study included 67 maternity health care clinics, 72 well-baby clinics, and 69 dental health care clinics. Of the 1,292 newborn children, 1,003 (90.8%) were included in this study. RESULTS: Preventive dental health care contributed to dental caries being halted in only 13.2 percent of those children who had enamel caries at 3 years of age. The dmft index did not increase in 22.6 percent of those children who had dentinal caries at 3 years of age. For all others, the disease became more severe. Toothbrushing habits of 3-year-old children were very consistent over the two years studied. Children were at a risk for caries when their mothers had nine years of basic education, when they already had plaque and caries at 3 years of age, and when the frequency of eating sweets increased the most during the two-year study period. CONCLUSIONS: Among 3-year-old children, plaque is an indicator of caries risk and therefore should be a key element in health education. Those children who already have evidence of caries at 3 years of age should be the target of preventive dental services because of their increased risk. PMID- 10390710 TI - Maryland adults' knowledge of oral cancer and having oral cancer examinations. AB - OBJECTIVES: This study sought to determine (1) knowledge of risk factors for oral cancer, (2) knowledge of signs and symptoms of oral cancers, and (3) factors associated with having had an oral cancer examination among 916 Maryland adults 18 years of age and older. METHODS: A statewide, random-digit dial, computer assisted telephone survey was conducted. The pretested instrument consisted of 32 questions that required 12 minutes to complete. RESULTS: Overall, level of knowledge about risk factors for and signs and symptoms of oral cancers was low; misinformation was high. Although 85 percent reported hearing about oral or mouth cancer, only 28 percent of the respondents reported having had an oral cancer examination. Of these, 20 percent had the exam during the past year--the recommended frequency for persons 40 years of age or older. In logistic regression analysis, adults more likely to have had an oral cancer examination included those who thought personal behavior causes more cancer than environmental factors; had more knowledge about risk factors for oral cancer; and were 40-64 years of age, white, and better educated than their counterparts (P < .05). The primary reasons for not having an exam were "no reason/didn't know I should" and "doctor/dentist didn't recommend." CONCLUSIONS: These results demonstrate a need for interventions designed to increase knowledge levels of risk factors for, signs, and symptoms of oral cancers and the need for oral cancer examinations; and to increase oral cancer examinations. PMID- 10390709 TI - Adult dental visits in California: successes and challenges. AB - OBJECTIVES: This study sought to estimate and characterize the proportion of California adults who visited a dentist in the preceding year and to identify reasons for not going. METHODS: In 1995, 4,029 adults were interviewed by telephone as part of the California Behavioral Risk Factor Surveillance System. Items included recentness of a dental visit, dental insurance status, and number of teeth lost due to disease. Persons who had not seen a dentist within the preceding year were asked the main reason they had not gone. RESULTS: In 1995, 65.9% of adults reported visiting a dentist in the preceding twelve months. Use of dental services was greater among persons aged 35 years or older (70.4%) than among those aged 18-34 years (58.4%) and among those with dental insurance (74.9%) than those without (54.4%). Dental visits were less likely among adults living at or below 200 percent of the federal poverty level, those with less than a high school education, and the edentulous. Reasons most commonly cited for not seeing a dentist were no perceived reason to go (37.2%), cost (30.7%), and fear (9.2%). CONCLUSION: Substantial variation in use of dental services exists among California's adults. Achieving equity in access and opportunity for disease prevention in this state may require expanded dental insurance coverage and serious efforts in oral health promotion. PMID- 10390711 TI - Incidence of orofacial injuries in high school sports. AB - OBJECTIVES: This study determined the incidence of orofacial injuries in athletes attending seven neighboring Minnesota high schools who participated in varsity soccer, wrestling, and basketball during the 1996-97 academic year. METHODS: Incidence was determined through athletes' written surveys and athletic trainer records. RESULTS: Survey response rates ranged from 86.3 percent to 94.0 percent among schools for all sports. The incidence rate of at least one orofacial injury per athlete in a season was 27.6 percent (SD = 20.2) in soccer, 72.3 percent (SD = 9.3) in wrestling, and 55.4 percent (SD = 23.9) in basketball. Ten percent of athletes sustained dental injuries. Fixed orthodontic appliances posed a higher risk for sustaining an injury in all sports. The games-to-practices ratios for injuries were 6.8 (soccer), 1.2 (wrestling), and 1.8 (basketball). Half of the athletes believed mouthguards prevent injuries; however, only 6 percent of the athletes reported mouthguard use. Athletic trainers reported eight orofacial injuries. CONCLUSIONS: The substantial rate of orofacial injuries among high school athletes participating in soccer, wrestling, and basketball needs to be minimized. Dentists should ask their adolescent patients routinely about sports participation. Policies should be developed to require school officials to report orofacial injuries, to inform athletes of their risk for orofacial injuries, and to consider mandated mouthguard use for these athletes. PMID- 10390712 TI - Prevalence of HIV-associated periodontitis and chronic periodontitis in a southeastern US study group. AB - OBJECTIVES: This study estimates factors associated with the prevalence of HIV associated periodontal diseases (HIV-P) and the severity and extent of periodontitis in HIV-infected adults from North Carolina (NC). METHODS: Data are derived from a cross-sectional study of HIV-infected adults (total n = 326, dentate n = 316) treated at the University of North Carolina Hospitals. Outcomes were a diagnosis of HIV-P and measures of probing pocket depth (PPD), recession (REC), and clinical attachment level (CAL). Immunosuppression was measured by peripheral blood CD4+ cells/mm3. RESULTS: In addition to persons with HIV (non AIDS), this study included 10 percent of the AIDS cases in North Carolina. Median age was 37 years (range = 19-67); 78 percent were male and 60 percent were black. Sixty-two percent of persons had a probing pocket depth > or = 5 mm; 46% had recession > or = 3 mm, and 66 percent had attachment level > or = 5 mm in one or more sites. Cases of HIV-P (n = 15) were rare. Persons taking HIV-antiretroviral medication were one-fifth (OR = 0.20; 95% CI = 0.07, 0.63) as likely to have HIV P as those not taking those medications, controlling for CD4+ cell counts. CONCLUSIONS: HIV-infected persons in this study group from North Carolina exhibited severe and extensive measures of adult periodontitis. A small proportion experienced a severe form of HIV-P, which was attenuated by antiretroviral therapy. PMID- 10390713 TI - Components of self-reported oral health and general health in racial and ethnic groups. AB - OBJECTIVE: The objective of this study is to investigate the validity of the General Oral Health Assessment Index (GOHAI), a self-reported oral health measure, when used in an all-age adult sample of Hispanics and African-Americans. METHODS: The study groups were 506 disadvantaged Hispanic and African-American adults who were recruited at low-cost medical and dental clinics. To explore the validity of the GOHAI in an all-age, ethnically diverse sample, principal component and principal factor analyses were conducted on the 12 items of the GOHAI and the 14 items constituting the MOS physical/social and mental health components. RESULTS: Four factors accounted for 40 percent of the total variance of health as measured by the MOS and GOHAI items: general health, physical/worry oral health, mental health, and social oral health. An association between socioeconomic variables and each subscale was found to be significant except for the physical/worry oral health subscale. CONCLUSIONS: This study confirms that the GOHAI is valid when used in younger and ethnically diverse samples. The findings also emphasize that oral health is distinct from general health and that the use of generic self-reported measures of health may miss important aspects of oral health that are valuable for dental health professionals. PMID- 10390714 TI - Oral/pharyngeal, laryngeal, and lung cancer discharge trends in Department of Veterans Affairs hospitals. AB - OBJECTIVE: This paper describes trends in oral and pharyngeal (O/P) cancer diagnoses in Department of Veterans Affairs (VA) hospitals from 1983 to 1993 and compares these trends to those of laryngeal and lung cancers. METHODS: The VA patient treatment file was used to identify unique hospital discharges from 1983 to 1993 having ICD-9-CM codes for O/P, laryngeal, and lung cancers. Descriptive statistics were tabulated to determine prevalence and distribution. Trends of change over time were analyzed using regression analyses of the percent rate on year. RESULTS: Between 1983 and 1993 the annual number of O/P cases among users of VA hospitals decreased from 4,983 to 3,298. Despite overall declines in O/P cancer discharges in VA, cancer of the pharynx, tongue, and salivary gland continues to increase. O/P cancer in younger persons also continues to increase in VA. Overall, laryngeal cancers significantly increased, while no significant change was associated with lung cancer. CONCLUSIONS: VA needs to evaluate the changes in the patterns of O/P cancer to ascertain whether this represents a shift in care from inpatient to outpatient care. VA should seek further information regarding these trends to better plan, implement, and evaluate programs to provide early diagnosis and treatment targeted to veterans. PMID- 10390715 TI - New trends in developmental neuroimaging in psychiatry. PMID- 10390716 TI - PET in child psychiatry: the risks and benefits of studying normal healthy children. AB - 1. Inclusion of children, particularly healthy control children, is a continuing debate. 2. Why involve children in PET research? The assumption is that the knowledge gained from such studies is critical for the advance of prevention and treatment of psychiatric illnesses. 3. What are the risks of PET procedures? Radiation exposure poses the most difficult problem. The assessment of this risk needs to separate the emotional reaction at the mention of "radiation" from the consideration of objective data of large studies of health hazards associated with low-level radiation exposure. 4. The assessment of the benefit/risk ratio is critical to the conduct of research, and requires the evaluation of risks according to the ambiguous definition of "minimal risk". PMID- 10390717 TI - Development of the human corpus callosum during childhood and adolescence: a longitudinal MRI study. AB - 1. Interest in the morphologic development of the corpus callosum (CC) during childhood and adolescence stems from adolescent changes in cognitive functions subserved by the CC, reports of CC anomalies for a wide variety of childhood neuropsychiatric illnesses, and controversy regarding sexual dimorphism. 2. Characterization of the normal developmental pattern of the CC is hindered by enormous variability of its size. This is especially problematic for cross sectional studies seeking to assess possible non-linear developmental curves. 3. To more accurately characterize developmental changes, a longitudinal brain magnetic resonance imaging study with subjects rescanned at approximately 2 year intervals was conducted resulting in 251 scans from 139 healthy children and adolescents. 4. Midsagittal area of the CC, especially the posterior regions, increased robustly from ages 5 to 18 years. 5. Although the genu of the CC was significantly larger in males there were no sex differences in mean area after adjustment for total cerebral volume and the growth patterns did not differ between sexes. 6. Analysis revealed a non-linear increase in the splenium, the most posterior region, with increases greatest in the younger years. 7. The results of this longitudinal study, in addition to confirming and extending previous cross-sectional reports, provide an increasingly accurate yardstick from which to assess pathological development. PMID- 10390718 TI - A mathematical model for the analysis of cross-sectional brain glucose metabolism data in children. AB - 1. The authors present here a 5-parameter developmental function designed to describe quantitatively the time-course of changes in glucose metabolism with age. This function consists of a plateau phase which is described by the rate of increase with age and the height of the plateau, followed by a decline phase characterized by the rate of decrease to adult levels. These two phases are separated by a distinct point in time, at which the transition between the two phases occurs. 2. The model is designed to fit published data showing that glucose metabolic rate in frontal cortex at birth is about 60% of adult values (mean adult value is 24 mumol/100 g/min) and increases to slightly less than triple adult value by age 3. The metabolic rate remains at this level until the age of approximately 8 years when it starts the decline to adult values. 3. A procedure is presented which allows the approximate computation of the 98% confidence contours in data space for the developmental function. The computation is based on the joint probability function obtained from the model covariance matrix. 4. The newly designed 5-parameter developmental function is well suited to describe maturational changes of glucose metabolism. Due to its excellent model identifiability and the interpretability of individual parameters, this function is better suited for description of maturational changes than the gamma function. Furthermore, this function may provide a useful framework for interpretation of other data sets during development. PMID- 10390719 TI - Corpus callosal signal intensity in treatment-naive pediatric obsessive compulsive disorders. AB - 1. Obsessive compulsive disorder (OCD) is increasingly recognized as a severe, highly prevalent and chronically disabling disorder, emerging during childhood in as many as 80% of cases. The authors previously found significant abnormalities in the region of the corpus callosum (CC) connecting ventral prefrontal cortex and striatum in pediatric OCD patients compared to controls that correlated significantly with OCD symptom severity. We speculated that this abnormality might reflect aberrant myelinization in OCD patients. 2. In order to better characterize the abnormality, the authors examined CC signal intensity (SI), believed to be a reliable index of myelinization of the CC. Lower numbers would indicate a greater concentration of white matter, while higher numbers indicate higher concentrations of gray matter. We compared the SI from midsagittal magnetic resonance images of 21 treatment-naive OCD patients, 7.2-17.7 years, and 21 case-matched healthy controls to examine regional CC signal intensity of the anterior, middle and posterior genu, body, isthmus, and the anterior, middle and the posterior splenii. 3. Mean total genu SI for the patient group (.993 + .006) was significantly less than the total genu SI of controls (.994 + .006) at F(1,37) = 4.73; p = .036. This abnormality in SI was localized to the CC region connecting ventral PFC and striatum, the anterior genu for the OCD group (.991 + .007) which was also less than control (.995 + .007) at F(1,37) = 5.47; p = .025., with no abnormality observed in middle or posterior genu regions. Genu SI was also inversely correlated with OCD symptom severity (r = -.55, p = .013) but not illness duration. Genu SI also correlated positively with genu area (r = .52, p = .020) in OCD patients but not controls. 4. Developmental abnormalities in genu size may arise from abnormalities in myelination in early onset OCD patients. The increased genu myelination observed in OCD patients may alter signal transduction and function of VPFC-striatal association circuits. PMID- 10390720 TI - An MRI study of the basal ganglia in autism. AB - 1. High-resolution MRI scans were obtained from 35 relatively high-functioning persons with autism and 36 healthy controls, comparable in age, gender, and IQ. 2. Volumetric measurements were obtained from manual tracing of the bilateral caudate, putamen, and globus pallidus. 3. An increased volume of the caudate nuclei was found in subjects with autism. Caudate enlargement was proportional to increased total brain volume in subjects with autism. 4. Caudate volume was associated with compulsions and rituals, difficulties with minor change, and complex motor mannerisms in autism. 5. Based on evidence of caudate abnormalities, a second MRI study was completed which replicated the finding of caudate enlargement in autism using an independent sample. 6. The caudate may be part of an abnormal distributed neural network in autism and involved in the ritualistic--repetitive behaviors of the disorder. PMID- 10390721 TI - Cerebellar vermis lobules VIII-X in autism. AB - 1. The aim was to investigate cerebellar vermis cross-sectional area in a group of high-functioning autistic children and normal control children. 2. Cerebellar vermis area measurements were completed on MRI scans from 8 autistic children (mean age 12.5 +/- 2.2, mean IQ 83.3 +/- 11.9) and 21 normal children (mean age 12.0 +/- 2.8, mean IQ 115 +/- 11). 3. The area of cerebellar vermis lobules VIII X was significantly smaller in the autistic children than in the normal control subjects. ANCOVA demonstrated a confounding effect of IQ on these results. 4. Larger studies of autistic and normal subjects will be needed to assess the relationship between cerebellar abnormalities, autistic symptoms and IQ. PMID- 10390722 TI - Evidence of altered energy metabolism in autistic children. AB - 1. In this pilot study, the authors investigated the hypotheses there are increased concentrations of lactate in brain and plasma and reduced brain concentrations of N-acetyl-aspartate (NAA) in autistic children. 2. NAA and lactate levels in the frontal lobe, temporal lobe and the cerebellum of 9 autistic children were compared to 5 sibling controls using MRS. Plasma lactate levels were measured in 15 autistic children compared to 15 children with epilepsy. 3. Preliminary results show lower levels of NAA cerebellum in autistic children (p = 0.043). Lactate was detected in the frontal lobe in one autistic boy, but was not detected any of the other autistic subjects or siblings. 4. Plasma lactate levels were higher in the 15 autistic children compared to 15 children with epilepsy (p = 0.0003). 5. Higher plasma lactate in the autistic group is consistent with metabolic changes in some autistic children. The findings of altered brain NAA and lactate in autistic children suggest that MRS may be useful characterizing regional neurochemical and metabolic abnormalities in autistic children. PMID- 10390723 TI - A SPECT HMPAO study of regional cerebral blood flow in depressed adolescents and normal controls. AB - 1. The objective of this study was to compare the relative regional cerebral blood flow (rCBF) patterns of a group of adolescents with major depressive disorder (MDD) to a group of normal controls. 2. Seven adolescent patients with symptomatic MDD and 7 age- and gender-matched normal controls, underwent SPECT imaging with 99mTc-HMPAO while unmedicated and in a resting state. These subject's data were normalized to whole brain counts, oriented in Talairach space, and analyzed using a voxel-based, t-image approach. 3. The authors found relative rCBF increases in the depressed group as compared to normals in the right mesial temporal cortex, the right superior-anterior temporal lobe, and the left infero-lateral temporal lobe. We found rCBF decreases in the depressed group as compared to normals in the left parietal lobe, the anterior thalamus and the right caudate. 4. Adolescents with MDD show rCBF abnormalities similar to those found in adult MDD rCBF studies. Further controlled studies with larger numbers of MDD subjects and normal age- and gender-matched controls are necessary before any definitive conclusions can be made from these findings. PMID- 10390724 TI - Brain organization for language in children, adolescents, and adults with left hemisphere lesion: a PET study. AB - 1. There is evidence for pronounced brain plasticity during postnatal maturation. The authors hypothesized that left-hemisphere lesion would be associated with greater than normal language participation of the right hemisphere and that atypical asymmetry of perisylvian language activations would be enhanced after lesion occurring in early childhood as compared to lesion occurring later in life. 2. Eleven patients with left-hemisphere lesion (aged 8-33 yrs.) and 9 normal adult comparison subjects were studied, using [15O]-water positron emission tomography. One patient group (N = 6) had early lesion onset (< or = 6 years of age), a second group (N = 5) had lesion onset later in life (> or = 10 years of age). Regional cerebral blood flow (rCBF) changes during listening to sentences (minus rest) and sentence generation (minus repetition) were compared between groups in predefined regions of interest. 3. Variance of regional activations within early and late lesion onset groups was considerable and qualitative inspection revealed only few robust group differences. However, when 4 patient pairs were approximately matched for chronological age, lesion site and VIQ, significantly reduced leftward asymmetry of activations in early lesion patients was found in the prefrontal, inferior frontal, and inferior parietal regions for expressive language, with concordant and marginally significant trends in the inferior frontal and superior temporal regions for receptive language. 4. The results suggest enhanced postlesional plasticity in childhood, while also reflecting strong individual variability probably due to clinical and demographic factors beside lesion onset. PMID- 10390725 TI - Functional organization of activation patterns in children: whole brain fMRI imaging during three different cognitive tasks. AB - 1. Patterns of brain activation were measured with whole brain echo-planar functional magnetic resonance imaging (fMRI) at 3.0 Tesla in healthy children (N = 6) and in one child with a left-hemisphere encephalomalacic lesion as sequellae from early stroke. 2. Three cognitive tasks were used: auditory sentence comprehension, verb generation to line drawings, and mental rotation of alphanumeric stimuli. 3. There was evidence for significant bilateral activation in all three cognitive tasks for the healthy children. Their patterns of activation were consistent with previous functional imaging studies with adults. 4. The child with a left-hemisphere stroke showed evidence of homologous organization in the non-damaged hemisphere. PMID- 10390726 TI - Central-amygdaloid carbachol suppressed nociceptive jaw opening reflex in freely moving rats. AB - 1. Experiments were carried out in rats with stimulating electrodes implanted in the dental pulp, recording electrodes inserted into the anterior digastric muscle, and indwelling cannula implanted in the central amygdaloid nucleus and the cisterna magna area. 2. Injection of 4.4 nM and 8.8 nM carbachol into the central amygdaloid nucleus suppressed digastric electromyogram (dEMG) to 81 +/- 8% and 47 +/- 9% of the control, respectively. 3. Atropine, a muscarinic receptor antagonist, blocked the suppression of dEMG in response to the administration of 8.8 nM carbachol into the amygdala. However, a mecamylamine, a nicotinic receptor antagonist, did not affect changes in dEMG. 4. Intracisternal naloxone, an opioid receptor antagonist, reduced the suppression of dEMG from 47 +/- 10 to 72 +/- 12% of the control. 5. Intracisternal methysergide, a serotonin receptor antagonist, also reduced the suppression of dEMG from 50 +/- 9 to 78 +/- 9% of the control. 6. The carbachol-induced antinociception from the central amygdaloid nucleus was attributed to opioid and serotonergic descending inhibitory influences on nociceptive pathways. PMID- 10390727 TI - Bradykinin and phorbol ester but not 5-HT2B receptor activation stimulate phospholipase D activity in the rat stomach fundus. AB - 1. Serotonin has been implicated as a mediator involved in migraine headache, an effect that may involve central 5-HT2B receptor activation. 5-HT2B receptor signal transduction in controversial. 2. Rat stomach fundus contraction to serotonin has been used as a model for 5-HT2B receptor activation. Serotonin induced contractility involves intracellular calcium release and activation of protein kinase C without stimulation of phosphoinositide (PI) hydrolysis. 3. Since phospholipase D (PLD) activation results in phosphatidic acid production, which can release intracellular calcium and provide diacylglycerol for PKC activation, the purpose of this study was to determine whether the 5-HT2B receptor coupled to PLD activation using the rat stomach fundus as a model system. 4. Using phosphatidylethanol production to measure PLD activity, both bradykinin (0.01-1 microM) and phorbol dibutyrate (PDBu, 1 microM) stimulated PLD activity in rat stomach fundal strips, indicating that this tissue possesses an active PLD system. 5. Under identical conditions, 5-hydroxytryptamine (5-HT) failed to stimulate PLD activity over a concentration range (1 nM-1 microM) documented to induce 5-HT2B receptor-mediated contraction in rat stomach fundus. Thus, the 5-HT2B contractile receptor in rat stomach fundus is not coupled to PLD activation, whereas both bradykinin and phorbol ester do couple to PLD. PMID- 10390728 TI - Neuropeptide Y Y1 receptor antagonist BIBP3226 produces conditioned place aversion in rats. AB - 1. Previous studies have shown that the blockade of the neuropeptide Y (NPY) Y1 receptors with N2-(diphenylacetyl)-N-[(4-hydroxy-phenyl)methyl]-D-arginine amide (BIBP3226) induces anxiogenic-like reaction in rats tested in elevated plus-maze test. 2. The present study examined whether such a treatment is aversive using place conditioning in a two-compartment apparatus. Locomotor activity was measured in open field test. 3. Pairings with potentially anxiogenic dose of BIBP3226 (5 micrograms/6.5 microliters, i.c.v.) produced a conditioned aversion for the drug-associated place, whereas the locomotor activity in the open field test was not affected by this dose of BIBP3226. 4. These data suggest that the blockade of central NPY Y1 receptors is aversive and provide additional evidence to the hypothesis that the NPY Y1 receptors are involved in the regulation of affective states. PMID- 10390729 TI - The effects of quinine and 4-aminopyridine on conditioned place preference and changes in motor activity induced by morphine in rats. AB - 1. The effects of two unselective potassium (K(+)-) channel blockers, quinine (12.5, 25 and 50 mg/kg) and 4-aminopyridine (1 and 2 mg/kg), on conditioned place preference and biphasic changes in motor activity induced by morphine (10 mg/kg) were tested in Wistar rats. Quinine is known to block voltage-, calcium- and ATP sensitive K(+)-channels while 4-aminopyridine is known to block voltage-sensitive K(+)-channels. 2. In the counterbalanced method, quinine attenuated morphine induced place preference, whereas 4-aminopyridine was ineffective. In the motor activity test measured with an Animex-activity meter neither of the K(+)-channel blockers affected morphine-induced hypoactivity, but both K(+)-channel blockers prevented morphine-induced secondary hyperactivity. 3. These results suggest the involvement of quinine-sensitive but not 4-aminopyridine-sensitive K(+)-channels in morphine reward. It is also suggested that the blockade of K(+)-channels sensitive to these blockers is not sufficient to prevent morphine-induced hypoactivity whereas morphine-induced hyperactivity seems to be connected to both quinine- and 4-aminopyridine-sensitive K(+)-channels. PMID- 10390730 TI - Neurotoxic effects of amphetamine plus L-DOPA. AB - 1. Male Swiss Webster mice were administered a series of amphetamine injections preceded by either saline or L-DOPA. 2. This injection regimen was performed for either one, two or three consecutive weeks and neurotoxic effects of the drugs were determined one week later. 3. Amphetamine treatment for two weeks produced a greater striata-dopaminergic lesion that treatment for only one week. Three weeks of treatment did not exacerbate the lesion, indicating that the damage had reached maximal levels. 4. L-DOPA pretreatment did not significantly alter any of the toxic effects of the amphetamine. Therefore, some dopaminergic neurons may be resistant to the toxic effects of amphetamine. PMID- 10390732 TI - Exercise in the rehabilitation of breast cancer survivors. AB - With the increase in the number of women who have survived breast cancer, there is a growing need to attend to the physical and emotional effects of cancer and its treatment as experienced by these survivors. Psychological distress, fatigue, weight gain, premature menopause and changes in body image are some of the long term sequelae of breast cancer. Exercise as an adjunctive treatment may help to attenuate these effects and thereby contribute to rehabilitation of women with breast cancer. We present data from the exercise literature and from studies on breast cancer patients that support this role of exercise. Following a critique of the research efforts, we present a brief outline of questions that should be addressed in evaluating the role of exercise in cancer rehabilitation. PMID- 10390733 TI - An exploratory study of social support: a cross-cultural comparison of Chinese-, Japanese-, and Anglo-American breast cancer patients. AB - This paper investigated the nature of social support for Asian- and Anglo American women post breast cancer treatment. Forty-six Anglo- and Asian-American (13 Anglo-American, 18 Chinese-American and 15 Japanese-American women) women were assessed 6 months to 3 years post-treatment. Assessments consisted of a semi structured interview plus standardized psychological tests. Three major hypotheses were developed and tested in the study. Results showed: (1) Anglo American women indicated a greater need for social support than either of the two Asian-American groups in 66% of the categories; (2) no differences were found between the three ethnic groups in receipt of emotional or tangible social support; and (3) the network size and composition differed significantly in 83% of the categories between the Anglo group and at least one of the Asian groups. These differences were in size, mode, and perceived adequacy of social support. Implications for culturally-based clinical practice which emerge from these findings are discussed. PMID- 10390731 TI - Brain concentrations of kynurenic acid after a systemic neuroprotective dose in the gerbil model of global ischemia. AB - 1. Kynurenic acid (KYNA) is a kynurenine metabolite and a broad spectrum excitatory amino acid antagonist that has been shown to be neuroprotective in models of cerebral ischemia, when administered exogenously. However, the actual concentration required in the CNS to evoke significant neuroprotection has never been assessed. 2. The purpose of this study was to address this question in the gerbil model of forebrain ischemia. KYNA (400-1600 mg/kg) or vehicle were administered i.p. 15 min before 5 min bilateral carotid occlusion. 3. Seven days after reperfusion, ischemia-induced hippocampal nerve cell loss (95% in vehicle treated) was significantly lower in KYNA-treated gerbils (65% and 52% at 1000 and 1200 mg/Kg, respectively, P < 0.01). Treatment with 1000 mg/kg produced brain KYNA concentrations that were dramatically elevated (135.9 and 42.3 microM in CSF and whole brain, vs 0.032 and 0.16 microM in controls, at 15 min after ischemia), as measured in a separate group of transcardially-perfused gerbils. Cerebral KYNA concentrations tended to return to basal values 2 hours after reperfusion. 4. These results indicate that KYNA has a marked neuroprotective effect in a model of forebrain ischemia. This activity is associated with KYNA concentrations in the brain and CSF that are compatible with the in vitro affinity of the compound for ionotropic glutamate receptors. PMID- 10390734 TI - The impact of a brief coping skills intervention on adherence to breast self examination among first-degree relatives of newly diagnosed breast cancer patients. AB - The present investigation sought to determine (1) the impact of a single session stress management/coping intervention (problem-solving training; PST) versus a general health counseling (GHC) control condition on breast self-examination (BSE) adherence among relatives of newly diagnosed breast cancer patients, and (2) whether women with heightened perceived risk of breast cancer and/or cancer specific distress at baseline were more likely to improve their BSE adherence following PST. The participants were 510 women age 20-75 who had at least one first-degree relative with breast cancer. All of the participants completed a baseline telephone interview, an intervention (PST versus GHC), and a 3-month follow-up telephone interview. The results revealed a 36% overall improvement in BSE adherence, with no significant between-group difference in improvement (chi 2 = 0.03, p = 0.87). The logistic regression analysis of improvement in BSE adherence revealed a statistically significant cancer-specific distress by treatment interaction (p = 0.04). Among women who received PST, those with high levels of cancer-specific distress were two times more likely to improve in BSE adherence than women low in cancer-specific distress. There was no effect of cancer-specific distress in the control condition. These results suggest that women with a family history of breast cancer who have high levels of distress may be most likely to benefit from behavioral coping skills intervention to promote adherence to breast cancer screening. PMID- 10390735 TI - Coping and adjustment to breast cancer. AB - This study examined possible predictors of adjustment to breast cancer. Sixty-one women participated soon after they were diagnosed with Stage I or Stage II breast cancer. Measures were gathered at diagnosis and again 4 months later. Predictor variables included aspects of the disease and treatment process and reported coping behavior. The most consistent predictor of distress and, to a lesser extent, quality of life, was avoidant coping: women who reported more avoidant coping were more distressed. These data fit well with most previous research and suggest one way of identifying women who may be more at risk for special difficulties coping with the diagnosis of breast cancer. PMID- 10390736 TI - Predictors of bereavement outcome for family carers of cancer patients. AB - AIMS: The psychological outcome of family carers after bereavement is an important issue in evaluating palliative care services. Palliative care services have the potential to provide preventive psychosocial intervention to family carers prior to bereavement, but are faced with the need to identify those who may have greatest risk of adverse outcome. This prospective study examines predictors of psychological outcome for family carers of cancer patients following bereavement based on factors identified at referral to a palliative care agency. METHODS: Cancer patients and their family carer were consecutively recruited and assessed on a range of clinical and psychological measures at referral to a palliative home care service in a metropolitan centre (Time 1). Carers were again assessed following the death of the patient, on average at 4 months post-bereavement (Time 2), using measures of bereavement symptoms and psychological morbidity. RESULTS: 178 carers were assessed on both occasions. The chief predictors of carer psychological symptoms and severity of grief at follow up were psychological symptom scores at the time of referral (Time 1). Factors also measured at Time 1 were significant predictors of symptoms and grief scores at Time 2: greater number of adverse life events, carer's coping responses, past bereavement and separation experiences, the relationship with the patient, and greater severity of patient's illness at the time of palliative care referral. CONCLUSIONS: The findings indicate clinical risk factors for adverse short-term bereavement outcome that can be identified in family carers during palliative care treatment, that have implications for identifying the psychological needs of carers, and that form a potential basis for interventions to enhance the psychological outcome for family carers. PMID- 10390737 TI - The Mental Adjustment to Cancer Scale--a psychometric analysis and the concept of coping. AB - A psychometric analysis of the Mental Adjustment to Cancer (MAC) scale was performed in a heterogeneous Swedish sample of cancer patients (n = 868). The homogeneity of the original subscales proved to be satisfactory (alpha coefficients 0.61-0.81). The sample was randomly split into two subgroups, and a factor analysis was carried out in one of them using the LISREL 8.20 procedure. This yielded four factors called 'Hopeless', 'Positive', 'Anxious' and 'Avoidant' including 28 of the 40 original items (alpha coefficients 0.58-0.81). The novel factor structure was cross-validated and confirmed in the second subgroup. In contrast to the original scale (one item), 'Avoidance', was indexed by three items. The distinction between mental adjustment and coping is discussed. It is concluded that both versions of the MAC scale are measures of mental adjustment including emotional reactions as well as coping. PMID- 10390738 TI - Collecting quality of life data in EORTC clinical trials--what happens in practice? AB - Problems with poor compliance when collecting quality of life data (QoL) in randomised clinical trials have prompted investigators to suggest measures to improve data collection. This study sought to look at the practical problems encountered by data managers and nurses in the cross-cultural setting of EORTC trials. A literature search was followed by a poster workshop session at a meeting of the EORTC study Group on Data Management and finally a postal questionnaire. The key problems identified centred around the lack of interest from some clinicians, lack of resources, inadequate protocols and a desire for training in the rationale for collecting QoL data to aid discussions with patients. Despite these problems many data managers and nurses found it rewarding to be on the 'frontline'. Since this study, the EORTC Data Center and in particular the Quality of Life Study Group and the Quality of Life Unit have implemented a number of measures to improve compliance. These include written guidelines for EORTC trials and a training course planned for Autumn 1999. PMID- 10390739 TI - Nicotine dependence and withdrawal in an oncology setting: a risk factor for psychiatric comorbidity and treatment non-adherence. AB - Highly nicotine dependent oncology patients are at high risk for psychiatric morbidity when they enter the medical care setting where smoking restrictions apply. Nicotine withdrawal symptoms exacerbate cancer-related distress as well as common physical side effects of cancer treatment. This case report illustrates the management of a patient whose ongoing treatment for bladder cancer was jeopardized as a result of nicotine dependence and withdrawal. Several associated complications are described, the most serious of which were his acute anxiety and non-adherence to medical recommendations. A short-term management approach that included anxiolytics and nicotine replacement was effectively used to reduce this patient's excessive anxiety and thus facilitate compliance with stressful treatments. The severity of complications that can result from untreated nicotine dependence and withdrawal underscores the importance of assessing and monitoring smoking status in every patient. Greater staff awareness of the clinical practice guidelines regarding the diagnosis and treatment of nicotine dependence will likely result in improved patient care and compliance. PMID- 10390740 TI - Anxiety symptoms and panic attacks preceding pancreatic cancer diagnosis. AB - Cancer of the pancreas is a highly malignant disease with a very poor prognosis. Depression and anxiety occur more frequently in cancer of the pancreas than they do in other forms of intra-abdominal malignancies and other cancers in general. Yet, the etiology of psychiatric symptoms in patients with cancer of the pancreas may not be traced solely to poor prognosis, pain, or existential issues related to death and dying. In as many as half of patients that go on to be diagnosed with the disease, symptoms of depression and anxiety precede knowledge of the diagnosis. This observation has raised speculation that mood and anxiety syndromes are related to disruption in one of the physiologic functions of the pancreas. In this paper, we present a patient who had no prior psychiatric history and developed panic attacks just prior to diagnosis of her cancer. To our knowledge, this is the first report in the literature where panic attacks, not simply anxiety, presented prior to a pancreatic cancer diagnosis. Her symptoms resolved following resection of the tumor. Implications of such phenomena for the diagnosis and treatment of anxiety and depression in pancreas cancer are discussed. PMID- 10390741 TI - Cunning but careless: analysis of a non-replication. PMID- 10390742 TI - The Fox guarding the clinical trial: internal vs. external validity in randomized studies. PMID- 10390743 TI - Rejoiner to Fox. PMID- 10390744 TI - [An experimental, randomized, double-blind, placebo-controlled clinical trial to infestigate the effect of nicardipine on cognitive function in patients with vascular dementia. Spanish group of nicardipine study in vascular dementia]. AB - INTRODUCTION AND OBJECTIVE: Experimental and clinical data suggest that calcium channel blockers could be effective in the treatment of vascular dementia (VD). The aim of this study is to evaluate the effect of nicardipine on the progression of cognitive impairment in patients with VD. PATIENTS AND METHODS: We selected outpatients, of both sexes, between 55 and 80 years old, with VD. Following a placebo wash-out period (4 weeks), the patients received nicardipine 20 mg t.i.d. or placebo for one year. The primary efficacy variable was the loss of over 10% of the basal score on the Mini-Mental State Examination after one year. The time it took to reach this degree of impairment, the evolution of Pfeiffer score, the effect on functional disability and the drug safety were also evaluated. RESULTS: 156 patients were included, 109 completed the study, and 142 underwent intent-to treat analysis. At the end of the study, 34.6% of patients with placebo and 21.1% with nicardipine had lost over 10% of their basal MMSE score. Favorable effects of nicardipine were found on females (40.9% vs 10.5%, p = 0.01876), patients without previous treatments (46.2% vs 13.3%, p = 0.00748) and patients with concomitant antiplatelet treatment (35.0% vs 15.9; p = 0.03836). Survival analysis showed that the patients with nicardipine took longer to lose cognitive capacities (p = 0.031; RR = 1.15-3.99). CONCLUSION: Nicardipine did not reduce the cognitive decline at the stated period in the total group but it significantly delayed this decline and produces better evolution in females, in patients without previous treatments, and those with antiplatelet treatment. PMID- 10390745 TI - [Symptomatic epilepsy: review of 208 patients]. AB - OBJECTIVE: To determine the main etiological mechanisms of symptomatic epilepsy and its frequency according to age. PATIENTS AND METHODS: We made a retrospective analysis of 208 patients admitted during a period of four and a half years, studying the variables: age, sex and type of seizures: simple partial, secondarily generalized partial, complex partial, tonic-clonic, generalized tonic, and also EEG and neuroimaging. RESULTS: The main etiological mechanisms found were: vascular (31.25%), alcoholic (12.01%), intracranial disorders (9.61%), traumatic (5.28%), degenerative (5.28%), infectious (2.88%) and cryptogenic (33.65%). In the last group there was an outstandingly large proportion of patients with silent infarcts. When considering vascular epilepsy, those seizures occurring during the acute phase of the stroke (24/65) are differentiated from those of late onset (41/65). In the latter there was a marked predominance of ischemic etiology (48.78% corresponded to extensive infarcts in the territory of the middle cerebral artery; 36.58% were associated with partial infarcts) probably because of the greater frequency of ischemic stroke as compared with hemorrhagic stroke. After the acute phase, the latency was of 10.68 +/- 0.43 months and the most frequent seizures were tonic-clonic (48.78%). CONCLUSION: In persons under 30 years of age, etiology is multifactorial; between 30 and 50 years of age alcoholic epilepsy (39.53%) and traumatic epilepsy (11.62%) predominate; over the age of 50 years the cause was vascular in 43.5%. In the latter age group there was a high proportion of patients with heraldic seizures. PMID- 10390746 TI - [Alpha interferon in the treatment of multiple sclerosis. Update and experience in Cuba]. AB - OBJECTIVES: We present an update of specific treatment for multiple sclerosis (MS), especially the form with a clinical course of exacerbation-remission (ER), in which we consider the benefits of beta interferons, copolymer 1 and intravenous immunoglobulin. We discuss the properties of alpha interferon and its modes of action which are very similar to those of beta interferon. The main clinical trials in which various subtypes of alpha interferons were used are summarized. PATIENTS AND METHODS: We establish the elements which justify interest in studying this substance and present the main trials carried out in Cuba. These consist in an initial trial in 9 patients, with encouraging results in the clinical course ER-MS and following this, the findings of a preliminary study of the first 17 patients of a randomized, double-blind National Clinical Trial, in which a placebo was used for control. CONCLUSIONS: In view of these results we recommend that the study being carried out in Cuba to confirm the efficacy of alpha interferon in the ER-MS form be continued. PMID- 10390747 TI - [Semantic verbal fluency in neurological patients without dementia with a low educational level]. AB - INTRODUCTION AND OBJECTIVE: The Semantic Verbal Fluency Test (sVFT) is very sensitive to cognitive deterioration. Standard values are usually those found in normal persons with an average or high cultural level. We analyze the results of a sVFT in a broad sample of persons assessed in a Neurology Clinic so as to find the standard values in this particular population. PATIENTS AND METHODS: A sVFT (animals in one minute) was given to 138 patients without dementia, aged over 55 years, assessed in the Neurology Clinic. Variables recorded were sex, age, years of schooling, studies completed, diagnosis and place of origin. A bivariate descriptive study and multivariate lineal regression analysis was done following a 'step by step' strategy. RESULTS: This group had a low educational level (72% had been to school for less than 10 years), with an average +/- standard error of 16.02 +/- 0.45 animals in one minute. The variables: years of schooling, sex, age and diagnosis showed a significant association with semantic verbal fluency in the adjusted regression model. CONCLUSIONS: Our values are lower than those of other standard groups. This may be related to the low educational level of our group and to the inclusion of persons with neurological disorders. The lineal regression model proposed permits prediction of the values of semantic verbal fluency in specific persons depending on their personal characteristics. PMID- 10390748 TI - [Spinal tumors in infancy. A report of 48 cases]. AB - INTRODUCTION: Spinal tumours in infancy are an infrequent oncological disorder. The clinical features, usually of insidious onset, are alterations in gait and a painful spine. OBJECTIVE: To analyze the different histological types of spinal tumours seen in infancy and their form of presentation in our series. PATIENTS AND METHODS: We reviewed the clinical histories of 48 patients with intraspinal tumours, aged under 15 years, whose reports of histological diagnosis had been sent to our centre. RESULTS: Of the patients studied, 17 were girls (35.4%) and 31 boys (64.6%) with an average age of 7.7 years. The histological diagnoses made most frequently were astrocytomas (22.9%) and lipomas (18.8%) followed by metastases (12.5%), ependymomas (8.3%) and Edwing's sarcoma (8.3%). The clinical features were present prior to diagnosis for between 1 and 6 months in 13 patients, and for less than one month in 9 patients. The presenting symptoms were alterations in gait and back pain in most patients. Exploratory tests were related to the involvement of long vias and second motor-neurone lesions. The main topographical findings were: in the axial plane the lesions were extradural (23 patients) and in the sagittal plane there was dorsal involvement (34 patients). CONCLUSIONS: Insidious, progressive alterations in gait together with continuous, nocturnal back pain are valuable data when a serious spinal disorder is to be suspected. Early diagnosis should be based on neuroimaging tests, essentially MR, in the patients in whom spinal cord involvement is considered. PMID- 10390749 TI - [A study of peripheral neural conduction, motor and sensory, in diabetic patients treated with hyperbaric oxygenation]. AB - INTRODUCTION: There are some occlusive disorders in the vasa nervorum and metabolic changes disminishing oxygen liberation by erytrocites at the capillary blood vessels, and these disturbances lead to endoneural microhypoxia. Hyperbaric oxygen reverts hypoxia in the diabetic neuropathy. OBJECTIVE: We studied motor and sensitive peripheric neuroconduction in nine diabetic patients, with distal symmetrical polyneuropathy, during normoglycemia. Four of them were insulin dependent and five were non insulin dependent. PATIENTS AND METHODS: The electrophysiological studies were done before treatment with hyperbaric oxygen, in a week, three and six months later. The abnormal electrophysiological parameters detected in diabetics were terminal latencies (enlarged), velocities of conduction (slowed) and distal amplitudes of compound action potentials (reduced). RESULTS: Neither distal latencies nor distal amplitudes and conduction velocities in peroneal nerve showed significative changes in the statistical analysis. We observed slower conduction velocities in the motor fibers of the median nerve in the examination performed six months after treatment. There was an increase of distal latency and retardation of the velocity of conduction six months later after treatment in the sensitive fibers of median nerve, whereas the amplitudes of sensitive action potentials decreased progressively. These changes suggest large diameter peripheral fibers didn't receive benefit with hyperbaric oxygen treatment. CONCLUSIONS: In all patients disappeared all symptoms of dysesthesias, paresthesias, distal pains and cramps in the legs and arms, suggesting functional changes in small unmyelinated fibers which we can't test with conventional techniques to prove it. PMID- 10390751 TI - [Akathisia, parkinsonism and depression induced by cinnarizine: a case report]. AB - INTRODUCTION: Cinnarizine is a calcium-entry blocker drug used in vertiginous disorders; among its most rare adverse effects appear extrapyramidal symptoms and depression, these effects can persist during weeks, months or years after the withdrawal of the drug and have been explained by the inhibition of the passage of calcium in striatal neurons and a direct antidopaminergic features because of the similar chemical structure with neuroleptic drugs. Clinical case. A case of cinnarizine-induced akathisia, parkinsonism and depression in a 25 years-old patient after 11 days of treatment is described. Sequential evaluation were done using the following instruments: Barnes' scale for akathisia, Simpson-Angus scale for extrapyramidal symptoms, Beck's depression scale, Zung's depression scale and SCID for major depression according to DSM-IV criteria. The patient was treated with benzodiazepines, propranolol and orphenadrine. CONCLUSIONS: Although cinnarizine-induced extrapyramidal symptoms and depression have been associated with old age and prolonged time of treatment, it must be considered its apparition among young patients and after a short time of treatment. PMID- 10390750 TI - [Response to high dose corticosteroids in a girl with bilateral optic neuritis]. AB - INTRODUCTION: Optic neuritis is rare in childhood. Frequently (35-52% of all cases depending on the series) they have, during their clinical course, foci of demyelination leading to the clinical picture of multiple sclerosis (MS). Since 1993, the optic neuritis study group has recommended treatment with high doses of corticosteroids, since this seemed to stop progression, improve long-term results and delay the appearance of MS. The course of our patient was better than we expected. CLINICAL CASE: A 10 year old prepubertal girl complained of progressive loss of vision and slight pain in the right eye for 26 days before admission to hospital. On examination there was obvious papillitis of the right ocular fundus with total loss of the pupillary light reflex, together with consensual hyporeflexia of the left eye. Study of the visual evoked potentials (VEP) showed that there was marked delay of the P-100 wave, and a lower amplitude in the right eye. Magnetic resonance imaging did not show any demyelinated focus. Serological testing for neurotropic viruses was negative. CONCLUSIONS: After the initial phase of intravenous treatment (third day) there was subjective recovery of vision and the pupillary light reflex returned. VEP studies showed marked recovery. Thirty days after treatment was started there was almost complete subjective and VEP recovery. This rapid progress, as compared to that of other paediatric cases published, suggests a mechanism involving decompression of the optic nerve. PMID- 10390752 TI - [Spinal cord ischemia as a consequence of dissection of an abdominal aortic aneurysm]. AB - INTRODUCTION: Ischemic spinal cord infarct is the most frequent vascular lesion, but although aortic aneurysms are a possible cause, it is unusual for such cases to be seen. Clinical case. We present a case of spinal ischemia as the first sign of the dissection of an aneurysm of the abdominal aorta. A 58 year old man was seen in the hospital Emergency Department complaining of lumbar pain and the sudden onset of paraplegia of the legs, associated with pain in the middle of his back but with no history of previous trauma or effort. The only relevant personal history was of smoking. Whilst he was in the Neurology Department, the anomaly was diagnosed after dorsal, and lumbar gadolinium magnetic resonance (MR), when a zone of ischaemia at T9-T10 was seen and, as a casual observation, an image compatible with an aneurysm of the abdominal aorta. The relationship between the dissection of the aorta and the neurological complications may be explained by a clear understanding of the vascular supply to the spinal cord. In this case, both the clinical findings and the MR were clearly indicative of an anterior spinal artery syndrome. CONCLUSIONS: In spite of its rarity, aortic aneurysm should be included in the differential diagnosis of a clinical picture of ischemic myelopathy, especially when there is lumbar and/or abdominal pain before the appearance of neurological symptoms. Spinal MR is important for this diagnosis. PMID- 10390753 TI - [Self-limited acute encephalopathy related to measles component of viral triple vaccine]. AB - INTRODUCTION: Neurological disorders secondary to the measles component of viral triple vaccine are not frequent. In spite of controversy regarding the cause, the clinical, diagnostic and legal implications are worth considering. CLINICAL CASE: We present the case of a 16 month-old baby with a clinical picture of self limiting acute encephalopathy characterized by cerebellar ataxia and alterations in behavior, accompanied by the clinical signs of attenuated measles. The negative results of complementary tests and an obvious time-relationship with a triple virus vaccination lead us to interpret the condition as being secondary to the measles component of the vaccine. CONCLUSIONS: We consider that although there is a low incidence of complications, the index of suspicion is also low, and even lower in cases with only minor neurological signs. It is therefore possible that such reactions are under-reported. PMID- 10390755 TI - [Neuro spheres as a source of cells for repair of the central nervous system]. AB - INTRODUCTION: Neurons and oligodendrocytes are terminally differentiated cells. This means that once they have differentiated from their precursor cells, they cannot proliferate. A direct consequence of this type of differentiation is that cell repair is impossible in areas where neurodegenerative disease have caused the death of neurons and oligodendroglia. Recently multipotential neuroepithelial precursors of the central nervous system (CNS) have been isolated and characterized in vitro. DEVELOPMENT: In this study we review the capacity for nervous repair of these neuroepithelial precursors from the neurobiological point of view. These cells, known as neuro-spheres can be cultivated, amplified and cryopreserved for subsequent transplanting. Already there are many studies showing how neuro-spheres maintain their capacity for differentiation in vivo and that they can reach certain localized areas of the CNS. CONCLUSIONS: From this review we conclude that through the study and manipulation of these neuro spheres, new goals in CNS repair may be achieved. PMID- 10390754 TI - [Hypnic headache: a new case]. AB - INTRODUCTION: Hypnic headache is a condition characterized by nocturnal episodes of headache which periodically waken the sleeping patient. They usually occur in persons over 55 years of age and are thought to be due to some type of disturbance of biological rhythm. CLINICAL CASE: A 70 year old woman, with no relevant past history, complained that during the previous 12-14 months she had been woken from sleep by episodes of headache. The headache was diffuse, non pulsatile and very intense. The duration was variable, from 15 to 45 minutes and usually disappeared without requiring analgesics. The pain was not associated with autonomic or ocular disorders, nausea, vomitting or focal neurological signs. Both general and neurological examinations were completely normal. No alterations were seen on cranial CT. Treatment with lithium was started and there was marked improvement in the frequency of headaches. CONCLUSIONS: Hypnic headache is an unusual disorder characterized by episodes of holocranial, or rarely hemicranial, headache of moderate-severe intensity and periodic occurrence. Typically the headache wakes the patient whilst he is asleep ('alarm clock') and there are no autonomic or neurological alterations of any type associated with the episode. The condition usually affects persons over 55 years of age, and it has been related to changes in biological rhythms. This presumed alteration of biological pacemakers has been based on the periodicity of the episodes and the response to lithium. PMID- 10390756 TI - [Pharmacological mechanisms of the treatment of dyskinesias in Parkinson disease]. AB - OBJECTIVE: The progression of Parkinson's disease (PD) and levodopa therapy leads to development of motor and psychic complications that cause serious limitations to the management of the advanced disease. DEVELOPMENT: This article reviews the current literature regarding epidemiological, clinical, pathophysiological and therapeutics of levodopa-induced dyskinesias (LID). CONCLUSIONS: 1) The most important risk factors for LID are the cumulated doses of levodopa, young age at onset and severity of PD. 2) Pathophysiological data include the nigrostriatal system denervation, the prolonged exposure to levodopa and the integrity of striatal eferences; in addition there are some alterations of dopamine receptors and other neurotransmitter systems. 3) Some pharmacological measures, different for each type of LID, and surgery (pallidotomy, pallidal stimulation, subthalamic stimulation) can be useful for the therapy of LID. PMID- 10390757 TI - [Value of the ictal video EEG recording in the presurgical evaluation of temporal lobe epilepsy. Semiology and EEG patterns]. AB - INTRODUCTION: The surgical treatment of epilepsy has gained increased acceptance in the last decade, especially for temporal lobe seizures. Its success is based upon a strict selection of the candidates, which combines clinical, neurophysiological, neuropsychological and neuroimaging criteria. DEVELOPMENT: The video-EEG recording of the seizures still plays an important role in this selection process, as it provides enough amount of interictal activity, demonstrates the existence of different seizure types and enables to perform an adequate electroclinical correlation. It also helps recognizing patients with concomitant non-epileptic events. The widespread use of ictal video-EEG has allowed the validation of several clinical signs, such as the dystonic posturing of the limb or the postictal speech disturbance among others, as being quite useful in the process of localization and lateralization. In fact, with the proper analysis of the ictal behavior it is possible to achieve confident lateralizing information in the majority of patients with temporal lobe epilepsy. CONCLUSIONS: This knowledge, gathered through the analysis of the semiology is complementary to that obtained with the simultaneous recording of the EEG. When a meaningful electroclinical correlation is possible, and it is also concordant with the data provided by the MRI and neuropsychological testing, a surgical procedure can be warranted without the use of intracranial electrodes. PMID- 10390758 TI - [A critical review of the current pathogenesis of multiple sclerosis and possible future trends]. AB - INTRODUCTION: Multiple sclerosis (MS) is a disorder in which the pathogenesis is not clearly understood, but where the disability caused and its progression determine the seriousness of the condition. At present, inflammation due to T lymphocytes is thought to be the cause of the clinical features and resulting disability. The main feature is demyelination, but it has not been possible to show by immunology studies, neuroimaging studies or therapeutic trials that the binomial inflammation-demyelination correlates with the disability. Therefore other possibilities of pathogenesis which provide a better explanation for the clinical findings in MS may be considered. DEVELOPMENT: We may start with the hypothesis of the existence of gliotoxic and neurotoxic factors in a patient with MS. These, by means of electrical inhibition mechanisms, autoimmune phenomena and loss of the normal neurone-oligodendrocyte-astrocyte relationship, or a non specific immune response may give place to inflammation with secondary demyelination on the one hand and to axonopathy with alterations in the mechanisms of remyelination due to alterations in the signals between the axon and the oligodendrocyte on the other. CONCLUSIONS: When these three factors are considered, a better explanation is found for the factors which determine the progression of the disability, exhaustion of the inflammatory response, deficient remyelination and the poor correlation between demyelination and the progression of the disease. At the same time, new possibilities for approaches to the treatment and investigation of MS appear. PMID- 10390759 TI - [The treatment of epilepsy in medical conditions, geriatric and pregnant patients]. AB - INTRODUCTION: Treatment with antiepileptic drugs in patients with medical conditions, old age or pregnancy is associated with concomitant pathology and the specific physiopathological changes of age or pregnancy. DEVELOPMENT: The difficulties of treatment in special conditions are related to changes in the pharmacokinetics of antiepileptic drugs, interaction with other medication and with concomitant treatment with drugs which may potentially cause convulsions. Also, the occurrence of a particular disorder or of pregnancy may lead to changes in the frequency of crises. Besides this, the side-effects of antiepileptic drugs may be more marked or more frequent in patients with several other conditions or in the elderly. CONCLUSION: The treatment of epilepsy in patients with associated medical disorders, old age or pregnancy requires knowledge of the behaviour of antiepileptic drugs and the physiopathology of these conditions. PMID- 10390760 TI - [Multiple sclerosis of late onset in Santiago de Cuba]. PMID- 10390761 TI - [GM1 syndrome, generalized infantile gangliosidosis. Presentation of two cases]. PMID- 10390762 TI - [Primary cerebral lymphoma in a patient with HIV infection on antiretroviral treatment]. PMID- 10390763 TI - [Wolf-Hirschhorn syndrome]. PMID- 10390765 TI - [Diabetes mellitus and intracranial stenosis]. AB - INTRODUCTION: The risk of having a cerebrovascular accident (CVA) is approximately 3 to 5 times greater in the diabetic population than in non diabetics. The physiopathological mechanisms of CVAs in patients with diabetes mellitus are not well-known, since few studies of this have been done. OBJECTIVES: To verify the hypothesis that intracranial arteriosclerosis, interpreted from findings of stenosis on intracranial Doppler studies, is greater in diabetics than in non-diabetics, independently of other vascular risk factors. PATIENTS AND METHODS: Patients were selected from the Register of Cerebrovascular Diseases of the Neurology Department of the Hospital de Santa Maria as having had an ischemic cerebrovascular accident (CVA)/transient ischemic accident (TIA), assessed by a colour codified vertebral carotid eco-Doppler and transcranial Doppler. Intracranial changes in the diabetic and non-diabetic groups of patients were compared with relation to the presence of risks such as hypercolestrolaemia, arterial hypertension and smoking. RESULTS: 384 patients were included, 71 were diabetic and 313 non-diabetic. The two groups were comparable with regard to average age and sex. Intracranial stenosis was approximately 3.13 (95% CI = 1.69 5.79) times more frequent in the diabetic population than in non-diabetics. This difference persisted independently of the presence or absence of other vascular risk factors. CONCLUSIONS: Diabetes mellitus is an important independent risk factor of alterations in the intracranial blood vessels and therefore of CVA. The intracranial circulation should be studied in these patients in view of the great frequency of intracranial stenoses and possible future improvements with therapeutic intervention. PMID- 10390764 TI - [Multiple vertebro-basilar infarcts and cardio-embolism]. AB - INTRODUCTION: Stroke characteristics do not inform much about its etiology, even if they can suggest a specific mechanism. We thought that multiple vertebrobasilar infarcts could be related with embolism, namely cardioembolism. PATIENTS AND METHODS: From a hospital prospective registry of stroke we retrieved 73 cases of acute non-lacunar vertebrobasilar infarcts, without previous episodes of stroke in any territory or vertebrobasilar transient ischemic accidents (TIA). Two groups were compared: patients with single cerebral posterior artery infarct (49) and patients with multiple vertebrobasilar infarcts (24), in respect to conventional risk factors for cerebrovascular disease, ancillary procedures performed, and associated pathologic conditions, as possible infarct pathogenesis. RESULTS: Proportions of risk factors and ancillary procedures performed were similar in both groups, except for hypercholesterolemia, which was more frequent in multiple infarcts, and for transcranial Doppler, which was performed more frequently in multiple infarcts. In multiple infarct group, cardioemboligenic pathology was more frequent, as were medium-high emboligenic risk cardiac diseases and atrial fibrillation, although the difference did not reach statistical significance. CONCLUSIONS: Our results support the hypothesis that multiple non-lacunar vertebrobasilar infarcts, from a first ever stroke event, suggest cardioembolic etiology, and recommend performing an exhaustive cardiac investigation. PMID- 10390767 TI - [Young-onset multiple sclerosis]. AB - INTRODUCTION: Young onset multiple sclerosis is an infrequent situation which may present with atypical symptoms and uncertain outcome. OBJECTIVE: Our aim was to assess the clinical presentation and course in young onset multiple sclerosis, and analyze eventual data which might be helpful in establishing its prognosis. PATIENTS AND METHODS: We have retrospectively reviewed the clinical protocols of 17 patients with young onset multiple sclerosis, defined as presentation of symptoms before 21 years. Diagnosis was made according to Poser's criteria including clinical features, magnetic resonance imaging, cerebrospinal fluid findings, and evoked potentials. RESULTS: The mean age at onset was 16.9 +/- 4.4 and median time to diagnosis was four weeks. The clinical course was relapsing remitting in 76.5% and secondary progressive in 23.5%. The mean annual exacerbation rate was 1.5 +/- 0.9 and median time to second exacerbation was 12 months. The actual Expanded Disability Status Scale score is 2.6 +/- 2 after a mean disease duration of 11.4 +/- 8.0 years. The correlation between the Expanded Disability Status Scale score and the mean disease duration was the only statistically significant result. CONCLUSIONS: These results are similar to other studies, namely, age at onset did not correlate with final neurological disability. However, we must emphasize that any primary progressive form was found in our study. We conclude that in young onset multiple sclerosis, progression is not dependent on the age of onset and does not necessarily lead to an unfavorable outcome. PMID- 10390766 TI - [Relationship between parenteral gangliosides administration and the Guillain Barre syndrome]. AB - OBJECTIVE: We carried out a case a case-control study to analyze the relationship between parenteral gangliosides administration and the Guillain-Barre syndrome. PATIENTS AND METHODS: We retrieved 64 patients with the diagnosis of Guillain Barre syndrome, and 148 controls. In cases and controls the proportion and 95% confidence intervals (CI) of subject receiving gangliosides, was calculated. The number of patients with the Guillain-Barre syndrome who needed ventilation or died was also calculated. RESULTS: Four of 36 patients (95% CI = 81-0.6), over 40 years, received gangliosides prior to Guillain-Barre syndrome. One of these patients was ventilated (95% CI = 25-2) and died. None of the controls less than 40 years old took gangliosides, while from the 108 over 40 (95% CI = 15-4) 9 received gangliosides. None developed signs suggesting Guillain-Barre syndrome. Although gangliosides were more often used in Guillain-Barre syndrome (OR = 1.75), the difference was not significant (95% CI = 4.82-0.69). CONCLUSION: The present work proves that in spite of the association of Guillain-Barre syndrome, with gangliosides intake, there is no statistical difference between this group of patients and control population. PMID- 10390768 TI - [Epilepsy and arteriovenous malformations]. AB - OBJECTIVE: To describe the characteristics of the patients with epilepsy, associated to arteriovenous malformation. PATIENTS AND METHODS: We introduced 11 patients assisted in 1997, at University Cajuru Hospital, with arteriovenous malformation associated to refractory epileptic seizures. RESULTS: Six men and five women, with age between 12 and 40 years old (27 years in mean), were interned with epileptic crises, 4 generalized tonic-clonic, 3 partial simple secondarily generalized, one with complex partial and one with simple partial that the complex developed with secondary generalization. CT showed temporal lesion in 3 patients, parietal in 3 patient, ventricular in 3, parietal fose and temporoparietal on one patient, respectively. MRI was accomplished in 5 patients presenting temporal lesion in 3 and temporo-parietal in 2. It was possible to accomplish the EEG in 8 patients, all with epileptiform activity. The arteriography showed arteriovenous malformations in all the patients, 3 of the which died. Eight patients were guided for surgical treatment evidencing cavernous angioma in 6 patient and hemangioma in 2. CONCLUSIONS: We observed that cortical and not cortical arteriovenous malformation associates to epileptic seizures and in the cases in which it is possible, the surgical treatment favors the control of the seizures. PMID- 10390769 TI - [The clinical relevance of serial determinations of plasma concentrations of carbamazepine and sodium valproate]. AB - INTRODUCTION: Serum concentrations of valproate (VPA) and carbamazepine (CBZ) were monitored in 66 epileptic out patients to determine then pharmacokinetic profiles. PATIENTS AND METHODS: Thirty-eight patients were receiving monotherapy, 17 with CBZ and 28 with VPA, and 28 patients were receiving VPA and CBZ. All were receiving standard (not controlled-release) CBZ or VPA preparations. Blood samples were obtained 2, 4, and 6 hours after the last dose. Serum concentrations were determined by gas chromatography. RESULTS: Mean serum concentrations of patients receiving CBZ were 8.2, 7.3 and 6.4 micrograms/ml for the 2, 4 and 6 hours samples, respectively. Mean serum concentrations of CBZ in the group receiving CBZ + VAP were 7.8, 8.4 and 7.8 micrograms/ml, respectively. Patients receiving VPA had mean serum concentrations of 92.8, 97.8 micrograms/ml, respectively. Those receiving CBZ + VPA had serum concentrations of 69.1, 70.6 and 55.8 micrograms/ml, respectively. Mean variation in the CBZ serum concentrations was 45% (range 7-91%) in patients receiving monotherapy and 31% (range 8-93%) in patients receiving CBZ + VPA. Mean variation in VPA serum levels of the patients receiving CBZ + VPA (56%) was greater than those of patients receiving CPA alone (38%). Subtherapeutic serum levels of either drug were detected in 35% of the patients. CONCLUSIONS: The large inter- and intraindividual variability of serum concentrations showed that a single measurement does not reflect the reality or CBZ and VPA clinical kinetics. The results confirm that three samples collected at 2, 4, and 6 hours after the last dose determine a clinically useful kinetic profile. PMID- 10390770 TI - [Epilepsy and cerebral tumor]. AB - OBJECTIVE: To determine clinical and pathological features with seizures associated brain tumors. PATIENTS AND METHODS: This was a retrospective study through of an Epilepsy Program Protocol where we studied fifty patients admitted at Hospital Universitario Cajuru of Curitiba, Brazil, in 1996-1997. RESULTS: We studied 36 males and 14 females, aged 6 and 81 years old (mean 40.5). Twenty six patients had tonic-clonic seizures, 13 had simple partial secondarily generalized, 8 had simple partial, 2 complex partial and one with simple partial progressing to complex partial seizure. CT showed parietal expansive lesion on 14 cases, frontal expansive lesion on 14 cases, frontoparietal on 5; intraventricular tumor, sella turcica, temporal temporoparietal and fronto temporo-parietal expansive lesion 2 on each case, and suprasellar lesion, centrum semiovale, cerebello-pontine angle, ventricular trigone, fronto-naso-etmoidal and brain stem 1 on each case. All patients were submitted to a biopsy and/or resection of the lesion. The principal brain tumors were meningioma in 30%, astrocytoma in 22%, glioblastoma multiform in 14%, oligodendroglioma in 4% and pituitary adenoma in 4%. PMID- 10390771 TI - [Work conditions of patients with controlled epileptic crises]. AB - OBJECTIVES: To study the employment conditions in a group of patient epileptic adults with control of its seizures, assisted in the University Hospital of Cajuru. PATIENTS AND METHODS: We studied 70 patients that we applied the specific protocol, obtaining data of its origin, profession and job situation. RESULTS: We examined 46 men and 24 women, with mean age 34 years old, where 63 of urban origin and 7 rural. All the patients were in treatment with antiepileptic drugs and without seizures or less than three seizures in the last twelve months. The largest group of patients (35) they met unemployed: 14 retired, 11 of the motivated by the disease, 10 worked as autonomous, 3 only studied and 3 were public employees. PMID- 10390772 TI - [Long-term efficacy and tolerance of topiramate in 44 children with resistant epilepsy]. AB - INTRODUCTION: Topiramate (TPM) is a new antiepileptic drug with multiple modes of action which should theoretically represent a wide therapeutic spectrum. However, there is still little clinical experience of its use in children with epilepsy. PATIENTS AND METHODS: TPM was given during a period of 14.8 +/- 15.4 months, at an average dose of 6.6 +/- 2.5 mg/kg/day to 44 children with resistant epilepsy. They included 21 children with Lennox-Gastaut syndrome, 14 with partial epilepsy, 7 with multifocal epilepsy, one with polymorphic epilepsy and one with electrical changes during sleep. RESULTS: When TPM was associated with their treatment, a response of > 50% reduction in epileptic crises was seen in 76% of the cases (85% with Lennox-Gastaut syndrome, 64% with partial epilepsy, 71% with multifocal epilepsy) and suppression of crises in 12% of the cases (5% with Lennox-Gastaut syndrome, 21% with partial epilepsy and 14% with multifocal epilepsy). The drug was well tolerated and only stopped because of side-effects in 4.5% of the cases. CONCLUSIONS: TPM is an antiepileptic drug with a broad therapeutic spectrum, good clinical efficacy in children and is well tolerated by them. PMID- 10390773 TI - [Miller-Fisher syndrome and cavernous angioma]. AB - INTRODUCTION: The Miller-Fisher syndrome is considered a variant of acute inflammatory demyelinating polyradiculoneuropathy, generally is a benign disorder. CLINICAL CASE: We describe a patient who has had, acutely, ataxia, areflexia and ophthalmoplegia. It has been electrophysiologically diagnosed as Miller-Fisher syndrome. The symptoms appeared after an infectious disease, namely sinusitis. The evoked potential studies suggested lesions at the brain stem and we found a cavernous angioma at MRI. CONCLUSION: We prepared a review of the literature published so far, showing that there is still much controversy about the physiopathology of this syndrome and the importance of the immunologic diagnoses. PMID- 10390774 TI - [Two myopathy cases]. AB - INTRODUCTION: Early diagnosis and therapeutic measures of frequent pathologies like hyperthyroidism made their neurological complications less frequent. In spite of well known, these neurological complications are some times forgotten. CLINICAL CASES: We describe two myopathy cases, which presentation was namely fatigue. The inespecificity of this complaint create aetiology and syndromatic diagnosis difficulties, and therapeutic decision. In first case, the symptoms were very gradual, what is usual in older patients, and became extremely incapacitate, depending on others for every daily activity. In the second case, a young patient with diplopia suggested myasthenia. Both cases were diagnosed with hyperthyroidism and a quick improvement occurred with proper medication. Based in these cases, a thyrotoxic myopathy brief comment is made, as in spite of being frequent, these cases presented diagnosis difficulties. PMID- 10390775 TI - [Brain stem ischemia in a boy with resistance to C activated protein and elevated lipoprotein A]. AB - INTRODUCTION: Activated protein C resistance is the most common hereditary coagulation abnormality. In the majority of cases it results from a point mutation Arg506-->Gln of the factor V gene, and characterized by a poor anticoagulant response to activated protein C. CLINICAL CASE: We report the clinical case of a 6-year-old obese boy, who presented with acute hemiparesis. A cerebral MRI revealed an area of infarction in the left hemiprotuberance. Further investigation identified activated protein C resistance (heterozygosity for factor V Leiden) and elevation in lipoprotein (a). His mother also had factor V Leiden mutation. Prophylaxis with acetylsalicylic acid was instituted with favorable evolution. CONCLUSIONS: This mutation, isolated, is usually asymptomatic, unless other risk factors coexist. Although venous thromboembolism seems to be the main clinical manifestation, recent reports consider that activated protein C resistance is also a risk factor for arterial thrombosis and stroke in children. We reinforce the need for systematic and thorough evaluation of etiology and risk factors in cases of stroke in children. PMID- 10390776 TI - [Isolated trigeminal neuropathy associated with a fatty tumor of Meckel's cavum]. AB - INTRODUCTION: In recent years there has been increasing interest in isolated trigeminal sensory neuropathy. We present a case with an unusual association of this neuropathy and a fatty tumour of Meckel's cavum. CLINICAL CASE: A 24 year old man consulted for occasional episodes (during the previous two months) of dysesthesia of the right maxillary region. These were self-limiting and lasted only a few minutes. There was no lacrimation, apparent trigger factor, conjunctival injection or reduction in level of consciousness. There were no abnormal findings either on general or on full neurological examination. On cranial CT there was no signal from the tip of the right petrous temporal bone, but no space-occupying lesion nor pathological uptake of contrast material. On cranial MR there was an extra-axial lesion in the superior part of the tip of the right petrous temporal bone of 2 x 2 cm, localized to Meckel's cavum and right cavernous sinus, with a small lobule in the right lateral part of the prepontine cisterna. CONCLUSIONS: Trigeminal sensory neuropathy has been described in association with different connective tissue disorders, infections of the central nervous system, vascular dilatations and very varied types of tumours, particularly meningioma. The commonest site for lesions related to this clinical condition is the posterior fossa. A tumour in Meckel's cavum is rarely found in relation to this diagnosis, and from our review of the literature, involvement of fatty tumours seems to be rare. PMID- 10390777 TI - [Acute sensory neuropathy associated with a varicella-zoster infection]. AB - INTRODUCTION: Sensory neuropathy is a clinical entity which has been considered to be found in relation to neoplasia, Sjogren's syndrome, long-term pyridoxine treatment, or to be idiopathic. CLINICAL CASE: We present the case of a 33 year old woman who developed acute sensory neuropathy after being diagnosed as having varicella. She had no previous history of weight loss or other signs of neoplasia, no dryness of the mucous membranes or history of arthritis and had taken no toxic substances or pyridoxine, thus ruling out other causes of sensory neuropathy. The acute varicella infection was apparent from the clinical characteristics and the presence of specific IgM in serum. CONCLUSION: The clinical signs, in the absence of other circumstances which might have been related, together with the presence of acute varicella-zoster virus infection seems to indicate that the two conditions were related. PMID- 10390778 TI - [Symptomatic sinus bradycardia associated with donepezil]. AB - INTRODUCTION: Donepezil is a drug which is being used more and more widely in mild-moderate Alzheimer's disease. In general, it is well tolerated and the side effects are basically cholinergic-dependent. Symptomatic disorders of cardiac rhythm associated with the use of donepezil are extremely unusual. CLINICAL CASE: We describe the case of an 81 year old patient with hypertensive cardiopathy, who developed sinus bradycardia, fainting and left cardiac failure three weeks after starting treatment with donepezil. When donepezil was stopped the sinus bradycardia disappeared, a 24 hour electrocardiographic holter showed no signs of sinus node disease and no episodes of this type occurred during the following six months. CONCLUSION: Symptomatic sinus bradycardia is a possible adverse effect of treatment with donepezil in Alzheimer's disease. PMID- 10390779 TI - [Lumbosacral plexopathy as a form of presentation of an aneurysm of the iliac artery]. AB - INTRODUCTION: The etiology of lumbosacral plexopathy is often due to compression. One of the less common causes of this is aneurysm of the iliac artery. However, 13% of the patients with this disorder initially have symptoms of plexus irritation or deficit. CLINICAL CASE: We describe the case of a 42 year-old-man, with no previous medical history, who complained of right-sided sciatica for the previous three months. On examination there were clinical signs of a lesion of the right lumbosacral plexus. On CT of the pelvis and MR of the lumbar spine there were images compatible with an aneurysm of the right iliac artery. This diagnosis was confirmed on arteriography. Treatment was surgical (aneurysmography and right iliofemoral by-pass). The pain disappeared and the motor deficit improved considerably. CONCLUSION: When a patient presents with lumbosacral plexopathy, an iliac aneurysm should be considered as a possible etiological factor. In such cases early diagnosis is essential, since surgical treatment will usually resolve the clinical problem. PMID- 10390780 TI - [Electromyographic response associated with capsular stroke]. AB - INTRODUCTION: Lesions of the pyramidal system are characterized by their effects on qualitative aspects of movement. One of the features of pyramidal defects is the presence of associated movements or synkinesis. CLINICAL CASE: We present a case with residual associated electromyographic (EMG) response in a patient who had had a capsular stroke 11 months previously. The patient presented with acute onset of a syndrome of purely motor right hemiparesis (PMH) which improved satisfactorily with complete functional recovery three months later. Eleven months after the acute episode, there was good functional recovery and neurophysiological examination showed the presence of EMG responses associated with the unaffected limb (left hand) when voluntary movements were made with the right hand. CONCLUSION: The appearance of qualitative alterations of movement associated with pyramidal syndromes and particularly the presence of associated EMG responses in the case of capsular infarct described, may reflect the different processes of reinnervation and functional reorganization which occur following the lesion and which are involved in recovery of motor function. PMID- 10390782 TI - [Cerebral mapping: methodology and applications in clinical neurology]. AB - INTRODUCTION: Cerebral mapping is a technique for the functional evaluation of the central nervous system which is currently available. DEVELOPMENT: Cerebral mapping is a considerable improvement on the data available from a conventional EEG. As with all complementary clinical tests its usefulness depends on a knowledge of its advantages and limitations. Cerebral mapping has several advantages--resolution in time, it is harmless, its functional significance, cost...--which make it very useful. Apart from this, the cerebral mapping has limitations which should be remembered. All in all, interpretation of the results obtained using this technique requires special training. CONCLUSION: In this study we review the methodological basis of cerebral mapping and its possible practical clinical use in neurology. PMID- 10390781 TI - [Magnetoencephalography: a new functional diagnostic technique for the neurosciences]. AB - INTRODUCTION AND OBJECTIVES: We present a review on the technical, methodological and clinical advances in the functional study of the brain by means of magneto encephalography. We look back the milestones of its historical development, through the work of the major research groups on this field and through our group's works and database (including doctoral thesis). Discussion on the neurophysiological and biomagnetism basis is provided as well as description of technical developments in superconducting detectors (SQUID, Superconducting Quantum Interference Device), signal processing, enhancement of noise-signal ratio and dipole modeling. DEVELOPMENT: The need for brain functional studies has led to newer imaging procedures (functional magnetic resonance, PET, SPECT, etc.). Their spatial and temporal resolution and invasivity are compared to that of magneto-encephalography. Current equipment, up to 306 whole-head channels, may accurately detect cortical and subcortical activity. Apart from the physiological activity, it may be applied to a number of conditions: epilepsy (ictal, interictal and presurgical); dementia, movement disorders, stroke, eloquent cortex delimitation prior to tumour or lesion resection; learning disabilities and foetal studies. CONCLUSIONS: Magnetoencephalography provides with an excellent temporal, very good spatial resolution, acquires in real-time, without references and minimal interference. It entails a great advance in the diagnostic approach in neurosciences. PMID- 10390784 TI - [Resistant vegetative state: considerations regarding bioethics of contemporary medicine]. AB - INTRODUCTION: The diagnosis and conduct toward persistent vegetative state (PVS) is one of the emergent themes in bioethic in our contemporary society. This clinical condition is defined. DEVELOPMENT: To homologate PVS with brain death (BD) is one of the most discussed present controversies at the bioethic international area. If we keep in mind the present concepts of BD, it's not correct to homologate both terms. There is an increasing practice in admitting the end of medical treatment in PVS. In fact, with the introduction of cost effectiveness concept in intensive medicine, the right to treatment of these patients is discussed at Intensive Care Units. Some present criteria about this are presented, taking age into consideration, diagnostic certainty and the establishment of function recovery prognosis. CONCLUSIONS: The introduction of recently developed models for the rehabilitation of patients with severe brain injuries and PVS may lead to substantial improvements in outcome and may also be cost efficient. It is not ethical to make an arbitrary decision to withdraw a medical treatment of a patient, when we know there is the structural possibility of recovering some functions. PMID- 10390783 TI - [Genetical aspects in cerebrovascular disease]. AB - INTRODUCTION: Stroke is a pathophysiological heterogeneous syndrome with numerous genetic influences. DEVELOPMENT: Previous populational studies had demonstrated the familial correlation. The parental history of stroke has been associated with stroke in the descendant with a significant relative risk of 2. A study of mono- and dizygotic twins found a fivefold risk of stroke among monozygotic compared with dizygotic. Other modifiable and non-modifiable risk factors are involve in cerebrovascular disease. Recently, the gene that codifies the angiotensin converting enzyme has been determine. An insertion/deletion polymorphism of was associated with increased levels of angiotensin converting enzyme. ApoE4 and high levels of homocysteine has been reported as a risk factors for cerebral infarction. Apoptotic mechanisms have recently been discovered in rats models of ischemia. Caspases inhibitors have shown a reduction of 40% in the cerebral infarction area. CONCLUSION: The purpose of this work is to analyze the genetic factors that constitute an influence in stroke. PMID- 10390785 TI - [Atherosclerosis and brain circulation]. AB - INTRODUCTION: Atherosclerosis affects the vascular system in a diffuse way and its is clearly implicated in some of the most prevalent diseases in western countries such as cerebrovascular and cardiovascular diseases. Knowing more about the underlying pathogenic mechanisms may contribute to a better understanding of this entities and the development of therapies for both its treatment and prevention. DEVELOPMENT: We review herein the concepts included in the term atherosclerosis, the growth of the atheromatous plaque and its complications and the cellular mechanisms which intervene in its development. We analyze how it influences brain hemodynamics and its implication in cerebrovascular ischemic disease paying attention to the dissimilarities with other vascular territories and the clinical syndromes which derive from its development on different vascular structures. CONCLUSIONS: Under the concept of ischemic cerebrovascular disease we can find a group of heterogeneous clinical syndromes, usually associated to different etiopathogenic mechanisms: cardioembolic, atherothrombotic or hemodynamic. Although their risk factors may be common, these processes are clearly different form each other. Therefore including ischemic brain infarctions all together without attending to their etiology may produce important methodological biases when interpreting the results in clinical trials or other studies, and may also be a suitable explanation for differences between authors. PMID- 10390786 TI - [A 78-year-old man with weakness of the limbs and skin lesions]. PMID- 10390787 TI - Pediatric health care and management. AB - Pediatric health care is an integral part of providing for the general health needs of puppies and kittens from birth to 6 months of age. Successful rearing of puppies and kittens requires providing them with a suitable environment; the correct quantities and quality of nutrients for growth; a regular schedule of feeding, sleeping, grooming, and exercise; and the stimulus that provokes micturation and defecation. The intestinal parasites, such as hookworms, roundworms, whipworms, tapeworms, Giardia, and Cryptosporidium, occur commonly in puppies and kittens. The advantages of early-age spay/neuter far outweight the risks. PMID- 10390788 TI - Fading kitten syndrome and neonatal isoerythrolysis. AB - Fading kitten syndrome includes noninfectious and infectious causes for neonatal death (birth to weaning age). Noninfectious causes are mostly responsible for mortality in the first week of life and include congenital disorders, low birth weights, trauma, malnutrition, environmental causes, and neonatal isoerythroylsis. Infectious causes are more prevalent at 3-4 weeks of age. This article discusses the causes, clinical signs, and management of fading kitten syndrome. PMID- 10390789 TI - Dental diseases of puppies and kittens. AB - Although there are certain differences warranting specific options therapeutically in young animals, many of the basic dental principles still apply. The key is examining the oral cavity of every patient at each visit and knowing how to recognize and manage any abnormalities encountered. Providing sound oral care from the time of a patient's first visit can figure significantly in a lifelong commitment to oral health that can have a positive impact on the patient's overall health. PMID- 10390791 TI - Pediatric dermatology. AB - There are a great many disorders that can affect young animals in addition to those with a hereditary component. Still, it is important to consider genodermatoses whenever a young animal presents with dermatological lesions. Fortunately, most dermatological conditions that affect young pups and kittens carry a good prognosis for diagnosis and full recovery. It is, however, important to correctly identify those animals with a poor prognosis if for no other reason than to offer supportive genetic counseling to minimize the risk of the trait being perpetuated. PMID- 10390790 TI - Congenital deafness and its recognition. AB - Congenital deafness in dogs and cats is primarily of the hereditary sensorineural form associated with white pigmentation genes, although acquired forms of deafness are possible. Highest prevalence is seen in white cats, especially those with blue eyes, and the Dalmatian, with many other dog breeds affected to some extent. This deafness results from degeneration of the cochlear blood supply at age 3-4 weeks, presumably resulting from suppression of melanocytes by the white (cat) or merle or piebald (dog) genes. Mechanism of inheritance is not understood for most breeds. Such animals should not be bred and may present liabilities for their owners. Objective diagnosis of deafness, especially when unilateral, relies on the brainstem auditory evoked response, an electrodiagnostic test where electrical activity in response to a click stimulus is recorded from the scalp using needle electrodes and a special purpose computer. Client counseling guidelines are presented. PMID- 10390792 TI - Recent information about hip dysplasia. AB - Dogs suffering from HD have a genetic background, but the releasing factors are many. Two of the most important are overnutrition and overexercising, especially in the young puppy. Radiography can give an estimate of the degree of secondary OA, and by evaluating the laxity in the hip joints at an early age, it seems to be possible to predict the chances for later development of OA. The eradication program has to be instituted in such a way that only the best dogs are accepted into breeding programs. It seems to be more efficient to eradicate the genetic part of the etiology by creating a breeding index by means of evaluating the offspring of the male dogs. PMID- 10390793 TI - Early spay-neuter in the dog and cat. AB - Early spay-neuter refers to the surgical sterilization of 8- to 16-week-old animals. Anesthetic and surgical techniques for the dog and cat are described. Pros and cons of prepubertal gonadectomy are discussed, and the veterinary literature is reviewed. PMID- 10390794 TI - Feline respiratory diseases. AB - Respiratory diseases in kittens can quickly result in life-threatening emergencies if not identified and managed early. Congenital anomalies of young cats are extremely uncommon. Viral respiratory infections in the cat are primarily caused by feline herpesvirus type 1 and feline calicivirus. Primary bacterial respiratory infections occur sporadically in cats. Bordetella bronchiseptica may cause severe respiratory signs in young kittens. PMID- 10390795 TI - Cardiopulmonary resuscitation and oxygen therapy. AB - Cardiopulmonary resuscitation and oxygen therapy are often necessary procedures done in veterinary practice. There are variations in CPR technique, especially in cardiac life support. Oxygen therapy can be an important adjunctive therapy in emergency and critical care medicine. The techniques used for oxygen administration differ depending on the medical problem and the animal. PMID- 10390796 TI - Pediatric intensive care. AB - To provide optimal care, a veterinarian in a pediatric intensive care situation for a puppy or kitten should be familiar with normal and abnormal vital signs, nursing care and monitoring considerations, and probable diseases. This article is a brief discussion of the pediatric intensive care commonly required to treat puppies or kittens in emergency situations and for canine parvovirus type 2 enteritis. PMID- 10390797 TI - Failure to grow. AB - Failure to grow in pups and kittens can be the result of many factors. Dietary, metabolic, endocrine, parasitic, neoplastic, and genetic diseases may be responsible for a failure to thrive alone or in concert with other disorders. A complete history, physical examination, complete blood cell count, biochemistry profile, and urinalysis are the initial steps to define the underlying disorder(s). Subsequent tests may be needed based on these initial diagnostic results. PMID- 10390799 TI - Welfare implications of sheep ear tags. AB - The damaging effects of ear tags used to identify sheep were studied by examining the ears of sheep after slaughter in three different abattoirs and the ears of sheep on a farm. In total, 1040 ears with tags were examined. There were six types of ear tag: metal 'Ketchum' style loop tags; two-piece rigid plastic tags; 'Allflex' style flexible plastic tags with a male and female part; golf tee shaped plastic ear tags; one-piece rigid plastic loop tags; and one-piece flexible plastic tags with a flap. The metal loop tags and plastic loop tags caused the most lesions, and the majority of the severe lesions. Ear tags placed near to the tip of the ear appeared to cause more damage. Some of the Ketchum style metal tags and two-piece rigid plastic tags appeared to be relatively new, as if recently fitted. These tags were more often associated with ear lesions, particularly moderate or severe lesions. The Allflex style flexible plastic tags caused the fewest problems, and the golf tee-shaped plastic tags also caused significantly fewer problems than the other tags. PMID- 10390798 TI - Pediatric ocular emergencies. AB - There are few ocular emergencies that are unique to the pediatric patient. Most ocular emergencies are traumatic in origin, and the prognosis is often determined by the extent of the injury. Some congenital anomalies that may present as ocular emergencies are also discussed. The focus of this article is recognition and initial therapy for the more common pediatric ocular emergencies. PMID- 10390800 TI - Pattern-matching models for the differential diagnosis of bovine spongiform encephalopathy. AB - This study assessed the performance of a system for making decisions about the diagnosis of bovine spongiform encephalopathy (BSE). The system consisted of four pattern-matching models. The sensitivity, specificity, likelihood ratios and accuracy of each model were determined by using clinical descriptions of 100 suspect BSE cases which had been submitted for brain histopathology by veterinary officers of the Ministry of Agriculture, Fisheries and Food, 50 of which were true positive cases (confirmed by histopathology) and 50 false positive cases (not confirmed by histopathology). The clinical description of each case consisted of 14 clinical signs, each of which was defined as either present or absent. The system compared the case descriptions with the profiles of possible differential diagnoses, each profile consisting of the frequency of occurrence of the same 14 clinical signs. The pattern-matching models used the sums of the sign frequencies to rank the differential diagnoses. Models 1 and 2 derived information only from the presence of signs; models 3 and 4 derived information from the presence and absence of signs. Models 2 and 4 excluded diagnoses which did not have in their profile a sign which was observed, and diagnoses which had a sign in their profile which should always be present according to the profile description but which was not observed. The best performances by the models were: sensitivity 96 per cent (model 1 and model 2), specificity 72 per cent (model 4), accuracy 72 per cent (model 4), likelihood ratio of a positive test 2.00 (model 4), likelihood ratio of a negative test 0.21 (model 4). PMID- 10390801 TI - Chronic pulmonary disease in West Highland white terriers. AB - This paper describes the clinical features, and diagnostic findings of a chronic respiratory condition in 29 West Highland white terriers. Typically, the dogs were coughing chronically, had dyspnoea and tachypnoea of varying severity, and had deteriorated progressively over months to years. The mean (sem) survival time in months from the clinical signs being first noted by the owners was 17.9 (2.3). Most cases had a combination of respiratory signs, but coughing was the predominant sign in 18 cases. Inspiratory crackles were audible on chest auscultation in 28 cases, 10 of which were also wheezing. Rhonchi were the predominant sound in the remaining case. The main radiographic changes were mild to severe increased Interstitial markings in all cases, with additional bronchial markings in 14 of the dogs. Right-sided cardiomegaly (cor pulmonale) was recorded in 15. Bronchoscopic findings in 17 of the dogs were either normal or involved a mild airway mucoid reaction in eight. Chronic mucosal changes were observed in eight, but in two this finding was equivocal. Dynamic changes to the lumen of the airway were present in seven cases. No significant haematological or biochemical changes could be detected in 20 cases, but four cases were hypercholestrolaemic. A histopathological assessment of four cases revealed alveolar septal fibrosis to be the predominant change. Prednisolone, with or without bronchodilators, was the most commonly used therapy, and the response was variable. The condition appears to be associated with significant pulmonary interstitial fibrosis of unknown aetiology and has clinical similarities to idiopathic pulmonary fibrosis (cryptogenic fibrosing alveolitis) in human beings. PMID- 10390802 TI - Transmissible gastroenteritis and porcine epidemic diarrhoea in Britain. PMID- 10390803 TI - Cytological and microbiological results from equine guttural pouch lavages obtained percutaneously: correlation with histopathological findings. PMID- 10390804 TI - Scouring in lambs following treatment with Vecoxan. PMID- 10390805 TI - Replacement of quarantine. PMID- 10390806 TI - Unrolling hedgehogs. PMID- 10390807 TI - Ectopic pregnancy in a rabbit. PMID- 10390808 TI - Preparation and properties of immobilized pig kidney aminoacylase and optical resolution of N-acyl-DL-alanine. AB - Aminoacylase (EC 3.5.1.14) was immobilized into DEAE-Sephadex A-25 by ion exchange absorption for optical resolution of N-acyl-DL-alanine. The effects of pH, temperature, and Co2+ concentration on the activity of free and immobilized enzymes were investigated along with the operational and the thermal stability of the immobilized enzyme. The immobilized enzyme retained high catalytic activity. The optimum pH and temperature for the hydrolysis of N-acyl-L-alanine in the DL isomer mixture were 8.0 and 65 degrees C, respectively. Co2+ was an activator for the immobilized enzyme in a similar role as for the free enzyme. No significant loss of activity was observed for at least 300 h of continuous operation. The yield of L-alanine was about 70% of the theoretical yield. The immobilized aminoacylase column decayed over a very long period of operation, but could be completely reactivated by regeneration. PMID- 10390809 TI - Anaerobic upflow fixed-film bioreactor for biomethanation of salty cheese whey. AB - In order to develop a suitable reactor for the biomethanation of high-strength salty cheese whey, the performance of anaerobic upflow fixed-film reactors packed with different support materials, such as charcoal, gravel, brick pieces, pumice stones, and PVC pieces, has been studied. The charcoal-bedded reactor gave the best performance, with the maximum gas production (3.3 L/L digester/d) and an enriched methane content (69% CH4). Temperature and hydraulic retention time were optimized, with the ultimate aim of improving biomethanation. Maximum gas production (3.3 L/L digester/d) was achieved at a hydraulic retention time of 2 d at 40 degrees C. PMID- 10390810 TI - Laccase from Coriolus hirsutus as alternate label for enzyme immunoassay. Determination of pesticide 2,4-dichlorophenoxyacetic acid. AB - A new label--laccase from the fungus Coriolus hirsutus--was applied for solid phase enzyme-linked immunosorbent assays of the pesticide 2,4 dichlorophenoxyacetic acid (2,4-D). Two proposed assays are based on (1) competitive binding of antibody-laccase conjugate with immobilized 2,4-D-protein conjugate and 2,4-D in tested sample, and (2) competition of 2,4-D and 2,4-D laccase conjugate for binding with immobilized antibodies. Kinetic and concentration dependencies for these reactions were studied, and the ELISAs were optimized in accordance with the data obtained. The elaborated systems permit the detection of 2,4-D in concentrations down to 10-20 ng/mL; time of the assays is 1.5-2 h. The main advantage of the laccase label, in comparison with the widely used peroxidase one, lies in the lack of hydrogen peroxide from substrate mixture, because dissolved oxygen plays the role of oxidizer. PMID- 10390811 TI - Stabilization and translation of immobilized mRNA on latex beads for cell-free protein synthesis system. AB - The stability of immobilized mRNA against ribonucleases was investigated in a cell-free protein synthesis system. The plasmid-encoding protein A with the 20 mer poly(A) tail under the control of T7 promoter was constructed, and the corresponding mRNA was synthesized by T7 RNA polymerase reaction. The resulting mRNA was immobilized on oligo(dT)-immobilized latex beads by hybridization utilizing the poly(A) tail of mRNA at the 3'-terminus. The mRNA was stabilized against three types of nucleases (3'-OH exonuclease, 5'-OH exonuclease, and endonuclease) by immobilization. Translation of immobilized mRNA with a continuous-flow cell-free protein-synthesizing system from Saccharomyces cerevisiae was ascertained. Reusability of the immobilized mRNA as genetic information was also examined. PMID- 10390812 TI - Partial purification and properties of putrescine oxidase from Candida guilliermondii. AB - Putrescine oxidase ([PO]; E.C. 1.4.3.4), which catalyzes the oxidative deamination of putrescine into gamma-aminobutyraldehyde, has been partially purified from Candida guilliermondii. Among the substrates tested, putrescine has the highest reaction rate, followed by spermidine and cadaverine. The K(m) values for putrescine, spermidine, and cadaverine were 20, 200, and 1.1 mM, respectively. The optimum pH and the temperature for PO were 8.0 and 37 degrees C, respectively. Growth of Candida species on putrescine as the sole nitrogen source induced the synthesis of PO that converts putrescine into delta 1 pyrroline and gamma-aminobutyric acid. These two products were detected and identified from the culture medium. The enzyme was not activated by divalent cations. Among the species of Candida tested, the highest enzyme activity was found in cell-free extracts of C. guilliermondii. The pathway of putrescine degradation was identified by substrate analysis to be along the nonacetylated pathway in C. guilliermondii. PMID- 10390813 TI - Recent trends in the biochemistry of surfactin. AB - The name surfactin refers to a bacterial cyclic lipopeptide, primarily renowned for its exceptional surfactant power since it lowers the surface tension of water from 72 mN m-1 to 27 mN m-1 at a concentration as low as 20 microM. Although surfactin was discovered about 30 years ago, there has been a revival of interest in this compound over the past decade, triggered by an increasing demand for effective biosurfactants for difficult contemporary ecological problems. This simple molecule also looks very promising as an antitumoral, antiviral and anti Mycoplasma agent. Structural characteristics show the presence of a heptapeptide with an LLDLLDL chiral sequence linked, via a lactone bond, to a beta-hydroxy fatty acid with 13-15 C atoms. In solution, the molecule exhibits a characteristic "horse saddle" conformation that accounts for its large spectrum of biological activity, making it very attractive for both industrial applications and academic studies. Surfactin biosynthesis is catalysed non ribosomally by the action of a large multienzyme complex consisting of four modular building blocks, called the surfactin synthetase. The biosynthetic activity involves the multicarrier thiotemplate mechanism and the enzyme is organized in structural domains that place it in the family of peptide synthetases, a class of enzymes involved in peptidic secondary-metabolite synthesis. The srfA operon, the sfp gene encoding a 4' phosphopantetheinyltransferase and the comA regulatory gene work together for surfactin biosynthesis, while the gene encoding the acyltransferase remains to be isolated. Concerning surfactin production, there is no indication whether the genetic regulation, involving a quorum-sensing mechanism, overrides other regulation factors promoted by the fermentation conditions. Knowledge of the modular arrangement of the peptide synthetases is of the utmost relevance to combinatorial biosynthetic approaches and has been successfully used at the gene level to modify the surfactin template. Biosynthetic and genetic rationales have been described for building variants. A fine study of the structure/function relationships associated with the three-dimensional structure has led to the recognition of the specific residues required for activity. These studies will assist researchers in the selection of molecules with improved and/or refined properties useful in oil and biomedical industries. PMID- 10390814 TI - Glucose overflow metabolism and mixed-acid fermentation in aerobic large-scale fed-batch processes with Escherichia coli. AB - Industrial 20-m3-scale and laboratory-scale aerobic fed-batch processes with Escherichia coli were compared. In the large-scale process the observed overall biomass yield was reduced by 12% at a cell density of 33 g/l and formate accumulated to 50 mg/l during the later constant-feeding stage of the process. Though the dissolved oxygen signal did not show any oxygen limitation, it is proposed that the lowered yield and the formate accumulation are caused by mixed acid fermentation in local zones where a high glucose concentration induced oxygen limitation. The hypothesis was further investigated in a scale-down reactor with a controlled oxygen-limitation compartment. In this scaledown reactor similar results were obtained: i.e. an observed yield lowered by 12% and formate accumulation to 238 mg/l. The dynamics of glucose uptake and mixed-acid product formation (acetate, formate, D-lactate, succinate and ethanol) were investigated within the 54 s of passage time through the oxygen-limited compartment. Of these, all except succinate and ethanol were formed; however, the products were re-assimilated in the oxygen-sufficient reactor compartment. Formate was less readily assimilated, which accounts for its accumulation. The total volume of the induced-oxygen-limited zones was estimated to be 10% of the whole liquid volume in the large bioreactor. It is also suggested that repeated excretion and re-assimilation of mixed-acid products contribute to the reduced yield during scale-up and that formate analysis is useful for detecting local oxygen deficiency in large-scale E. coli processes. PMID- 10390815 TI - Metabolic responses to different glucose and glutamine levels in baby hamster kidney cell culture. AB - In this work, a BHK21 clone producing a recombinant antibody/cytokine fusion protein was used to study the dependence of cell metabolism on the glucose and glutamine levels in the culture medium. Results obtained indicate that both glucose and glutamine consumptions show a Michaelis-Menten dependence on glucose and glutamine concentrations respectively. A similar dependence is also observed for lactate and ammonia productions. The estimated value of the Michaelis constant for the dependence of lactate production on glucose (KLacGlc) was 1.4 +/ 0.1 mM and for the dependence of ammonia production on glutamine (KAmmGln) was 0.25 +/- 0.11 mM and 0.10 +/- 0.03 mM, at glucose concentrations of 0.28 mM and 5.6 mM respectively. At very low glucose concentrations, the glucose to lactate yield decreased markedly, showing a metabolic shift towards lower lactate production. This metabolic shift was also confirmed by the significant increase in the specific oxygen consumption rate also observed at low glucose concentrations. Although it was highly dependent on glucose concentration, the oxygen consumption also increased with the increase in glutamine concentration. At very low glutamine concentrations, the glutamine to ammonia yield increased, showing a more efficient glutamine metabolism. PMID- 10390816 TI - A new route to L-threo-3-[4-(methylthio)phenylserine], a key intermediate for the synthesis of antibiotics: recombinant low-specificity D-threonine aldolase catalyzed stereospecific resolution. AB - A new enzymatic resolution process was established for the production of L-threo 3-[4-(methylthio)phenylserine] (MTPS), an intermediate for synthesis of antibiotics, florfenicol and thiamphenicol, using the recombinant low-specificity D-threonine aldolase from Arthrobacter sp. DK-38. Chemically synthesized DL-threo MTPS was efficiently resolved with either the purified enzyme or the intact recombinant Escherichia coli cells overproducing the enzyme. Under the optimized experimental conditions, 100 mM (22.8 g l-1) L-threo-MTPS was obtained from 200 mM (45.5 g l-1) DL-threo-MTPS, with a molar yield of 50% and a 99.6% enantiomeric excess. PMID- 10390818 TI - Substrate specificities of the chloromuconate cycloisomerases from Pseudomonas sp. B13, Ralstonia eutropha JMP134 and Pseudomonas sp. P51. AB - The chloromuconate cycloisomerase of Pseudomonas sp. B13 was purified from 3 chlorobenzoate-grown wild-type cells while the chloromuconate cycloisomerases of Ralstonia eutropha JMP134 (pJP4) and Pseudomonas sp. P51 (pP51) were purified from Escherichia coli strains expressing the corresponding gene. Kinetic studies were performed with various chloro-, fluoro-, and methylsubstituted cis,cis muconates. 2,4-Dichloro-cis,cis-muconate proved to be the best substrate for all three chloromuconate cycloisomerases. Of the three enzymes, TfdD of Ralstonia eutropha JMP134 (pJP4) was most specific, since its specificity constant for 2,4 dichloro-cis,cis-muconate was the highest, while the constants for cis,cis muconate, 2-chloro- and 2,5-dichloro-cis,cis-muconate were especially poor. The sequence of ClcB of the 3-chloro-benzoate-utilizing strain Pseudomonas sp. B13 was determined and turned out to be identical to that of the corresponding enzyme of pAC27 (though slightly different from the published sequences). Corresponding to 2-chloro-cis,cis-muconate being a major metabolite of 3-chlorobenzoate degradation, the kcat/K(m) with 2-chloro-cis,cis-muconate was relatively high, while that with the still preferred substrate 2,4-dichloro-cis,cis-muconate was relatively low. This enzyme was thus the least specific and the least active among the three compared enzymes. TcbD of Pseudomonas sp. P51 (pP51) took an intermediate position with respect to both the degree of specificity and the activity with the preferred substrate. PMID- 10390817 TI - Functionality of biphenyl 2,3-dioxygenase components in naphthalene 1,2 dioxygenase. AB - Naphthalene 1,2-dioxygenase (Nap dox) and biphenyl 2,3-dioxygenase (Bph dox) are related enzymes that have differentiated during evolution as their specificity has changed. Although their component arrangement is similar, the structure of each component has been modified quite extensively. The purpose of this work was to determine the catalytic capacity of purified Nap dox toward chlorobiphenyls and to investigate the functionality of Bph dox components in the Nap dox system. Both enzyme systems were purified by affinity chromatography as histidine-tagged fused proteins. Data show for the first time that Nap dox can catalyze the oxygenation of all three monochlorobiphenyl isomers, but it is unable to hydroxylate 2,5-, 2,2'-, 3,3'-, 4,4'-di- and 2,2',5,5'-tetrachlorobiphenyl. The rates of cytochrome c reduction by the ferredoxin components of the two enzymes were identical when the Bph dox reductase component was used in the assay, showing an efficient electron transfer between the Bph dox reductase component and the Nap dox ferredoxin. However, when the Bph dox ferredoxin was used to reconstitute a hybrid Nap dox, the enzyme was only 22% as active as the parental enzyme. These data are discussed in terms of the potential use of Nap dox for the development of enhanced chlorobiphenyl-degrading dioxygenases. PMID- 10390819 TI - Development of a bioconversion process for production of cis-1S,2R-indandiol from indene by recombinant Escherichia coli constructs. AB - Recombinant Escherichia coli cells expressing the toluene dioxygenase (TDO) genes from Pseudomonas putida convert indene to cis-1S,2R-indandiol, a potentially important intermediate for the chemical synthesis of the HIV-1 protease inhibitor, Crixivan. A bioconversion process was developed through optimization of medium composition and reaction conditions at the shake-flask and 23-1 fermentor scales. A cis-1,2-indandiol productivity of approx. 1000 mg/l was achieved with construct TDO123, which represents a 50-fold increase over the initial titer. Varying the bioconversion conditions did not change the enantiomeric excess (e.e.) for the 1S,2R enantiomer from about 30%, suggesting that toluene dioxygenase intrinsically converts indene to 1S,2R- and 1R,2S indandiols at a ratio of 2:1. Further inclusion of the Pseudomonas dehydrogenase gene in construct D160-1 led to the production of chirally pure cis-1S,2R indandiol (e.e. > 99%) as a result of the selective degradation of the 1R,2S enantiomer, with the overall yield (650 mg/l) proportionally reduced. A single stage process was developed for D160-1 and scaled up to the 23-1 fermentor, achieving a cis-1S,2R-indandiol titer of 1200 mg/l. PMID- 10390820 TI - Construction of a flocculent Saccharomyces cerevisiae fermenting lactose. AB - A flocculent Saccharomyces cerevisiae strain with the ability to express both the LAC4 (coding for beta-galactosidase) and LAC12 (coding for lactose permease) genes of Kluyveromyces marxianus was constructed. This recombinant strain is not only able to grow on lactose, but it can also ferment this substrate. To our knowledge this is the first time that a recombinant S. cervisiae has been found to ferment lactose in a way comparable to that of the existing lactose-fermenting yeast strains. Moreover, the flocculating capacity of the strain used in this work gives the process several advantages. On the one hand, it allows for operation in a continuous mode at high cell concentration, thus increasing the system's overall productivity; on the other hand, the biomass concentration in the effluent is reduced, thus decreasing product separation/purification costs. PMID- 10390821 TI - High-level expression of a recombinant protein in Klebsiella planticola owing to induced secretion into the culture medium. AB - The Tn5-based transposon Tn5-KIL3 (Miksch et al. 1997c) bearing the kil gene of the ColE1 plasmid of Escherichia coli, which mediates controlled export of periplasmic proteins into the culture medium, was stably integrated into the chromosome of Klebsiella planticola with high transposition frequency. A Bacillus hybrid beta-glucanase located on an RSF1010-derived plasmid was mobilized from E. coli to K. planticola and used as a reporter protein to select strains with high expression and secretion competence. During fermentation experiments it was shown that the production of beta-glucanase in K. planticola was improved to an unexpectedly high level when the enzyme was secreted into the medium. Due to the stationary-phase promoter used for the expression of the kil gene the secretion of beta-glucanase into the medium started at the transition from the exponential to the stationary phase, as in E. coli, and the fraction of secreted protein reached 90%. The results showed that K. planticola may represent an interesting organism for the production of heterologous proteins. PMID- 10390823 TI - Characterisation of a starch-hydrolysing enzyme of Aspergillus niger. AB - A UV-induced mutant strain of Aspergillus niger (CFTRI-1105-U9) overproduced a starch-hydrolysing enzyme with properties characteristically different from the known amylases of the fungus. The purified enzyme of 4.0 pI had an apparent molecular mass of 125 kDa and it dextrinised starch and then saccharified the dextrins. Patterns of the enzyme activity on starch, resulting in glucose at 60 degrees C and glucose, maltose and maltodextrins at 70 degrees C as primary products, suggested significant applications for the enzyme in starch-processing industries. PMID- 10390822 TI - The role of the alternative respiratory pathway in the stimulation of cephalosporin C formation by soybean oil in Acremonium chrysogenum. AB - Addition of soybean oil to Acremonium chrysogenum cultures growing on sugars doubled the specific production of cephalosporin C during the idiophase of growth. While the addition of soybean oil had no effect on the total rate of respiration, the respiration that proceeded via the alternative, cyanide insensitive pathway exhibited a more than twofold increase. Addition of soybean oil also stimulated the formation of isocitrate lyase activities. Inhibition of oxidative metabolism of one of the products of isocitrate lyase (succinate) by thenoyltrifluoroacetone completely inhibited the alternative respiratory pathway. The role of soybean-oil-stimulated alternative respiration in the stimulation of cephalosporin C production and the role of isocitrate lyase are discussed. PMID- 10390824 TI - Fermented whey--an inexpensive feed source for a laboratory-scale selenium bioremediation reactor system inoculated with Thauera selenatis. AB - It is critical that an inexpensive electrondonor/carbon-source be found for selenium bioremediation using the selenate-respiring bacterium, Thauera selenatis. Since acetate is a preferred substrate for growth of this organism, a method was developed for fermenting the lactose in whey to large amounts of acetate. Indigenous whey microorganisms fermented the whey lactose in this manner when grown in continuous culture at a very slow dilution rate (D = 0.05 h-1). The successful use of the fermented whey lactose as the carbon-source/electron-donor feed for a laboratory-scale selenium-bioremediation reactor system, inoculated with T. selenatis, treating selenium-contaminated drainage water was also demonstrated. Selenium oxyanions and nitrate were reduced by 98%. PMID- 10390825 TI - Genetic and morphometric characterization of mussels (Bivalvia: Mytilidae) from mid-atlantic hydrothermal vents. AB - Mussels were collected from deep-sea hydrothermal vents along the Mid-Atlantic Ridge. Specimens from the Snake Pit site were previously identified genetically and anatomically as Bathymodiolus puteoserpentis, but the relationships of mussels from other sites (Logatchev and Lucky Strike) were unclear. Molecular genetic and morphological techniques were used to assess differences among these mussel populations. The results indicate that the range for B. puteoserpentis extends from Snake Pit to Logatchev, and that an unnamed second species, B. n. sp., occurs at Lucky Strike. Analysis of mitochondrial NADH dehydrogenase subunit 4 (ND4) revealed 13% sequence divergence between the two species. Nei's genetic distance (D) based on 14 allozyme loci was 0.112. A multivariate morphometric analysis yielded a canonical discriminant function that correctly identified individuals from these sites to species 95% of the time. PMID- 10390826 TI - Identification of sibling species of the bryozoan Bugula neritina that produce different anticancer bryostatins and harbor distinct strains of the bacterial symbiont "Candidatus Endobugula sertula". AB - Although the cosmopolitan marine bryozoan Bugula neritina is recognized as a single species, natural products from this bryozoan vary among populations. B. neritina is the source of the anticancer drug candidate bryostatin 1, but it also produces other bryostatins, and different populations contain different bryostatins. We defined two chemotypes on the basis of previous studies: chemotype O contains bryostatins with an octa-2,4-dienoate substituent (including bryostatin 1), as well as other bryostatins; chemotype M lacks bryostatins with the octa-2,4-dienoate substituent. B. neritina contains a symbiotic gamma proteobacterium "Candidatus Endobugula sertula," and it has been proposed that bryostatins may be synthesized by bacterial symbionts. In this study, B. neritina populations along the California coast were sampled for genetic variation and bryostatin content. Colonies that differ in chemotype also differ genetically by 8% in the mitochondrial cytochrome c oxidase subunit 1 (CO I) gene; this difference is sufficient to suggest that the chemotypes represent different species. Each species contains a distinct strain of "E. sertula" that differs at four nucleotide sites in the small subunit ribosomal RNA (SSU rRNA) gene. These results indicate that the chemotypes have a genetic basis rather than an environmental cause. Gene sequences from an Atlantic sample matched sequences from the California chemotype M colonies, suggesting that this type may be cosmopolitan due to transport on boat hulls. PMID- 10390827 TI - Planetary environments and the origin of life. PMID- 10390828 TI - Prebiotic synthesis on minerals: bridging the prebiotic and RNA worlds. PMID- 10390829 TI - Molecular origins and the null hypothesis: motifs from our maker? PMID- 10390830 TI - The genomic tag hypothesis: modern viruses as molecular fossils of ancient strategies for genomic replication, and clues regarding the origin of protein synthesis. PMID- 10390831 TI - New evidence for the genomic tag hypothesis: archaeal CCA-adding enzymes and tDNA substrates. PMID- 10390832 TI - Archaeal aminoacyl-tRNA synthesis: unique determinants of a universal genetic code? PMID- 10390833 TI - Origins and evolution of AIDS viruses. PMID- 10390834 TI - Horizontal gene transfer: pitfalls and promises. PMID- 10390835 TI - Morphological transitions of brain sphingomyelin are determined by the hydration protocol: ripples re-arrange in plane, and sponge-like networks disintegrate into small vesicles. AB - The phase transition of hydrated brain sphingomyelin occurs at around 35 degrees C, which is close to the physiological temperature. Freeze-fracture electron microscopy is used to characterize different gel state morphologies in terms of solid-ordered and liquid-ordered phase states, according to the occurrence of ripples and other higher-dimensional bilayer deformations. Evidently, the natural mixed-chain sphingomyelin does not assume the flat L beta, phase but instead the rippled P beta, phase, with symmetric and asymmetric ripples as well as macroripples and an egg-carton pattern, depending on the incubation conditions. An unexpected difference was observed between samples that are hydrated above and below the phase transition temperature. When the lipid is hydrated at low temperature, a sponge-like network of bilayers is formed in the gel state, next to some normal lamellae. The network loses its ripples during cold-incubation, which indicates the formation of a liquid-ordered (lo) gel phase. Ripples re appear upon warming and the sponge-like network disintegrates spontaneously and irreversibly into small vesicles above the phase transition. PMID- 10390836 TI - Phase behavior of fluorocarbon and hydrocarbon double-chain hydroxylated and galactosylated amphiphiles and bolaamphiphiles. Long-term shelf-stability of their liposomes. AB - This paper describes the morphological characterization, by freeze-fracture electron microscopy, and the thermotropic phase behavior, by differential scanning calorimetry and/or X-ray scattering, of aqueous dispersions of various hydroxylated and galactosylated double-chain amphiphiles and bolaamphiphiles, several of them containing one or two hydrophobic fluorocarbon chains. Colloidal systems are observed in water with the hydroxylated hydrocarbon or fluorocarbon bolaamphiphiles only when they are dispersed with a co-amphiphile such as rac-1,2 dimyristoylphosphatidylcholine (DMPC) or rac-1,2-distearoylphosphatidylcholine (DSPC). Liposomes are formed providing the relative content of bolaamphiphiles does not exceed 20% mol. Most of these liposomes can be thermally sterilized and stored at room temperature for several months without any significant modification of their size and size distribution. The hydrocarbon galactosylated bolaamphiphile HO[C24][C12]Gal forms in water a lamellar phase (the gel to liquid crystal phase transition is complete at 45 degrees C) and a Im3m cubic phase above 47 degrees C. The fluorocarbon HO[C24][F6C5]Gal analog displays a more complex and metastable phase behavior. The fluorinated non-bolaform galactosylated [F8C7][C16]AEGal and SerGal amphiphiles form lamellar phases in water. Low amounts (10% molar ratio) of the HO[C24][F6C5]Gal or HO[C24][C12]Gal bolaamphiphiles or of the single-headed [F8C7][C16]AEGal improve substantially the shelf-stability of reference phospholipon/cholesterol 2/1 liposomes. These liposomes when co-formulated with a single-headed amphiphile from the SerGal series are by far less stable. PMID- 10390837 TI - Synthesis of acid-labile diplasmenyl lipids for drug and gene delivery applications. AB - The low pH environments characteristic of endosomal compartments and ischemic tissues provide an intrinsic pathway for triggering site-specific contents release from appropriately designed delivery vehicles. Accordingly, research in this group has focused on the design, synthesis and application of novel acid sensitive lipids that will undergo facile lamellar (L alpha) to hexagonal (HII) phase transitions within these acidic sites. Previously, it has been demonstrated that plasmenylcholine-type lipids have excellent acid hydrolysis and contents release kinetics (Gerasimov et al., Biochim. Biophys. Acta. 1324 (1997) 200-214; Rui et al., J. Am. Chem. Soc. 120 (1998) 11213-11218). This paper describes the synthesis of three new acid sensitive lipids, based on a chiral 1,2-di-O-(1Z',9Z' octadecadienyl)-sn-glycerol (6) platform, displaying phosphocholine (7), poly(ethyleneoxide) (8), and O-carbamoyl-N-diethylen-etriamine (10) headgroups. Intermediate 6 was obtained in 28% overall yield via a six step synthesis from (S)-(+)-2,2-dimethyl-1,2-dioxolane-4-methanol. Subsequent conversion to the final products was acheived in moderate (7 and 10) to excellent yields (8). PMID- 10390838 TI - Spirotox--a new tool for testing the toxicity of volatile compounds. AB - A new method for estimating the toxicity of volatile compounds was developed. The test was carried out in the disposable polystyrene multiwells. After the organisms, protozoa Spirostomum ambiguum, were added to the wells, microplate was tightly closed using silicone grease and polyethylene film. The toxicities of 21 organic compounds were estimated. No control mortality was observed in all cases. Transparent PE film enabled good observation of test response. The toxicity of tested compounds varied over 4 orders of magnitude. Deformations were 2-4 more sensitive toxic response then lethality. The toxicity of tested compounds in Spirotox test correlates well with the log Kow and toxicity results from other bioassays: Microtox, D. magna and T. pyriformis. PMID- 10390839 TI - An 'inherent' biodegradability test for oil products: description and results of an international ring test. CONCAWE Biodegradation Task Force. AB - Current test guidelines for assessing 'inherent' (potential) biodegradability were designed for water-soluble, organic compounds of low volatility and are unsuitable for most oil products. It was against this background, that CONCAWE (the oil companies' European organisation for environment, health and safety) formed a task force to develop a standard test protocol for assessing the 'inherent' biodegradability of oil products. PMID- 10390840 TI - Using probabilistic neural networks to model the toxicity of chemicals to the fathead minnow (Pimephales promelas): a study based on 865 compounds. AB - We investigate the use of probabilistic neural networks (PNN) to model the acute toxicity (96-hr LC50) to the fathead minnow (Pimephales promelas) based on a 865 chemicals data set. In contrast to most other toxicological models, the octanol/water partition coefficient is not used as input parameter. The information fed into the neural network is solely based on simple molecular descriptors as can be derived from the chemicals' structures and indicates the potential of this approach as general methodology for the estimation of toxicological effects of chemicals. PMID- 10390841 TI - Prediction of the fish acute toxicity from heterogeneous data coming from notification files. AB - Four descriptors (Molecular weight, log(Pow), hardness and free energy of solvation) were selected to predict, on a training set of heterogeneous chemical compounds, the fish acute toxicity. The data were extracted from 523 notification files of new chemicals stored at the French Department of the Environment. The selection of the descriptors was carried out by using a statistical technique coupling OLS regression and genetic algorithm. The limits of validity for the final equation are discussed by comparing the actual and predicted activities on several compounds. PMID- 10390842 TI - A comparison between toxicity tests using single species and a microbial process. AB - In this study the sensitivity of the acetate mineralization process performed by five strains of microorganisms in soil for the toxicants Zn2+ or PCP was calculated from the sensitivity of the contributing species. The species used were a fungus (Aspergillus niger CBS 121.49), an actinomycete (Streptomyces lividans 66), two Gram-negative Pseudomonas putida strains (MT-2 and DSM 50026) and a Gram-positive strain Rhodococcus erythropolis A177. For zinc the EC10 of the process performed by the five strains together was 77 mg/kg whereas for pentachlorophenol it was 2 mg/kg. The EC10 of the process was compared with the EC50 of the most sensitive species contributing to the process. P. putida MT2 was the most zinc sensitive strain (EC50 = 22 mg Zn/kg) and A. niger was the most sensitive strain for pentachlorophenol (EC50 = 1.4 mg/kg). This shows that a 10% inhibition of a process can be accompanied by a more than 50% inhibition of the most sensitive species. PMID- 10390843 TI - The toxicity of antibiotic agents to the luminescent bacterium Vibrio fischeri. AB - Despite their common use the fate and effects of antibiotics in the environment are largely unknown. These compounds may enter the environment through different pathways, resulting in the contamination of waste water or fresh water, where bacteria are most likely the primarily affected organisms. In this paper the toxicity of several drugs, reflecting the most important groups of antibiotics and chemotherapeutics, towards Vibrio fischeri are presented. The chronic bioluminescence inhibition assay with Vibrio fischeri is shown to be sensitive against many of the high volume antibiotics used for veterinary purposes and in aquaculture. Thus the assay may be a valuable tool for an effects assessment and biomonitoring of these xenobiotics. The available data for both parts of the risk assessment procedure--exposure assessment and effects assessment--have to be regarded as insufficient for most antibiotics. When the available data about environmental concentrations of antibiotics are compared with their toxicity towards Vibrio fischeri, direct effects on natural microbial communities are to be expected. PMID- 10390844 TI - Applicability of a yeast oestrogen screen for the detection of oestrogen-like activities in environmental samples. AB - A (xeno)oestrogen bioassay was introduced, using a genetically modified yeast strain which produces a fusion protein encompassing the human oestrogen receptor hormone binding domain and the yeast GAL4-DNA binding domain. Upon binding of appropriate substances this fusion protein recognises the respective DNA sequence thereby enhancing the transcription of a beta-galactosidase reporter gene. The bioassay procedure was evaluated by screening 30 compounds, including some known or suspected (xeno)oestrogens and determining EC50-values for 17 beta-oestradiol, 1.5 nM, 4-tert.-octylphenol, 6.7 microM and bisphenol A, 104 microM. Toluene extracts from different environmental matrices were tested for their oestrogenic activity. The positive test results obtained with a sewage sludge extract indicated the applicability of this bioassay for environmental monitoring. PMID- 10390845 TI - A new enzyme immunoassay for PCDD/F TEQ screening in environmental samples: comparison to micro-EROD assay and to chemical analysis. AB - A new Enzyme ImmunoAssay (EIA) for PCDD/F TEQ measurement in extracts of environmental samples was described. The bioassay TEQ which derived from EIA and EROD were compared with each other and with results from chemical analysis. For all environmental samples, the EROD-TEQ is higher than the value from chemical analysis. However, the EIA-TEQ is much more identical with the value from chemical analysis. Our results indicate that the EIA assay is a complementary method to the EROD assay and should be useful as a rapid and sensitive screening tool for environmental samples in many situations. PMID- 10390846 TI - Tissue-specific heavy metal (Cd, Pb, Cu, Zn) deposition in a natural population of the zebra mussel Dreissena polymorpha Pallas. AB - The zebra mussel Dreissena polymorpha was tested as an indicator of heavy metal exposure in urban waters of Vienna, Austria. Mussels, two sediment fractions, suspended matter, and filtrate were collected over one annual cycle at five sampling stations. Dreissena was dissected into five body parts: viscera, gill, foot, byssus, and shell, to determine tissue-specific metal accumulation. Cadmium, lead, copper, and zinc were measured by AAS. There was no clear relationship between (relatively low) metal concentrations in ambient compartments and in zebra mussel bodies. Therefore, D. polymorpha must be regarded as a poor suitable indicator tool under near-background contamination situations. Tissue-specific metal accumulation showed that cadmium was mainly stored in soft body parts. Lead, copper, and zinc showed significantly highest concentrations in the byssal threads. Metal concentrations and distribution patterns within the mussel's body must be interpreted as a result of (unknown) internal metal treatment/regulation. Excretion of lead, copper, and zinc via the byssus complex probably is an effective strategy for preventing toxic injury in D. polymorpha. PMID- 10390847 TI - Quantitative structure-toxicity relationships for halogenated substituted benzenes to Vibrio fischeri, using atom-based semi-empirical molecular-orbital descriptors. AB - Quantitative structure-toxicity relationships (QSTR's) are derived for an extensive series of halogenated benzenes, anilines, phenols, nitrobenzenes, toluenes and other substituted benzenes against Vibrio fischeri using a wide range of whole molecule and atom-based descriptors derived from semi-empirical molecular-orbital calculations. In terms of direct statistical correlation with toxicity it was found that the molar refractivity was the most important parameter, closely followed by the solvent accessible surface area of the compound. The accuracy of these descriptors in fitting the numerous fluoro- and chloro-mono-aromatic compounds was compared with bromine and iodine analogues, where the 'best' descriptors for the former were found in general to be less accurate for the latter in the case of multi-halogen substitution. The equations obtained were also used to classify the compounds into narcosis-based mechanisms of toxicity and those with respiratory uncoupling potential. A combination of the molar refractivity and the nucleophilic susceptibility of one of the meta ring carbons predicted the toxicity of the halo-benzenes and toluenes, along with anisoles, benzonitriles, nitrobenzenes and most of the anilines. The relevance of these descriptors to developing coherent and more generally applicable models for QSTR's of mono-aromatic compounds to other species in environmental toxicology is discussed. PMID- 10390848 TI - Environmental behavior of explosives in groundwater from the Milan Army Ammunition Plant in aquatic and wetland plant treatments. Removal, mass balances and fate in groundwater of TNT and RDX. AB - Phytoremediation of 2,4,6-trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro 1,3,5-triazine (RDX) in groundwater using constructed wetlands is a potentially economical remediation alternative. To evaluate Explosives removal and fate was evaluated using hydroponic batch incubations of plant and substrate treatments with explosives-contaminated groundwater amended with [U-14C]-TNT or [U-14C]-RDX. Plants and substrates were collected from a small-scale wetland constructed for explosives removal, and groundwater originated from a local aquifer at the Milan Army Ammunition Plant. The study surveyed three aquatic, four wetland plant species and two substrates in independent incubations of 7 days with TNT and 13 days with RDX. Parent compounds and transformation products were followed using 14C and chemical (HPLC) analyses. Mass balance of water, plants, substrates and air was determined. It was demonstrated that TNT disappeared completely from groundwater incubated with plants, although growth of most plants except parrot feather was low in groundwater amended to contain 1.6 to 3.4 mg TNT L-1. Highest specific removal rates were found in submersed plants in water star-grass and in all emergent plants except wool-grass. TNT declined less with substrates, and least in controls without plants. Radiolabel was present in all plants after incubation. Mineralization to 14CO2 was very low, and evolution into 14C-volatile organics negligible. RDX disappeared less rapidly than TNT from groundwater. Growth of submersed plants was normal, but that of emergent plants reduced in groundwater amended to contain 1.5 mg RDX L-1. Highest specific RDX removal rates were found in submersed plants in elodea, and in emergent plants in reed canary grass. RDX failed to disappear with substrates. Mineralization to 14CO2 was low, but relatively higher than in the TNT experiment. Evolution into 14C-volatile organics was negligible. Important considerations for using certain aquatic and wetland plants in constructed wetlands aimed at removing explosives from water are: (1) plant persistence at the explosives level to which it is exposed, (2) specific plant-mass based explosives removal rates, (3) plant productivity, and (4) fate of parent compounds and transformation products in water, plants, and sediments. PMID- 10390849 TI - Clearance of PCDD/Fs via the gastrointestinal tract in occupationally exposed persons. AB - A digestive tract mass balance was performed on six men with high body burdens of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs). Intake via food was measured by analyzing duplicate portions of the food consumed by the volunteers and excretion via feces was determined by quantitative collection and analysis of the feces. Blood samples were taken to determine the current body burden. The results showed that the quantity of non-metabolized chemical excreted in the feces clearly exceeded the uptake via food for all of the 2,3,7,8 substituted PCDDs and some of the PCDFs, indicating a significant clearance across the gastrointestinal tract. The concentrations of these PCDD/F congeners in blood and feces were highly correlated (r > 0.8), demonstrating that the fecal PCDD/F content was determined by the body burden. The half lives in the test persons due to fecal clearance of non-metabolized chemical were estimated from the excretion rate and the current body burden and ranged between 10 years (Cl8DD) and 33 years (2,3,4,7,8-Cl5DF). These were compared with the overall contaminant half-lives due to all clearance processes which were calculated from the body burden and the decrease in blood concentrations measured over several years. The fecal clearance of non-metabolized PCDD/F contributed on average between 37% (2,3,7,8-Cl4DD) and 90% (Cl8DD) to the total elimination. This indicates that the gastrointestinal pathway plays a decisive role in the clearance of most 2,3,7,8-subsituted PCDD/F congeners. PMID- 10390850 TI - Leaching of mecoprop and dichlorprop in calcareous soil. Effect of the exogen organic matter addition in this process. AB - Leaching studies of mecorprop (R,S)-2-(4-chloro-2-methylphenoxy)propanoic acid, and dichlorprop, (R,S)-2-(4-chloro-2,4-dichlorophenoxy) propanoic acid, under saturated conditions were conducted in unamended and amended soil columns. The purpose of the study was to investigate the leaching of these herbicides in three type of soils and the exogen organic matter effect on this process. The leaching patterns could be related to variation in the soil texture and diffusion processes of the herbicides into micropores within the walls of conducting pore. The leaching rate in the amended soil columns decreased with the addition of organic matter. The breakthrough curves (BTC) of these herbicides in the leachates of the amended soil columns were wider and more diffused than the BTC obtained for the corresponding unamended soil. The theoretical BTC overestimated the pore volume required for the displacement of these pesticides from the soil column. This may be due to the differences in the adsorption process between the bacth and soil columns methods. PMID- 10390851 TI - Deactivation of mycotoxins. I. An in vitro study of zearalenone adsorption on new polymeric adsorbents. AB - This study describes the elimination of zearalenone concentrations in vitro using two new polymeric forms of cross-linked polyvinylpyrrolidone (cryogels of cross linked PVP). Adsorption of zearalenone was studied under isothermal conditions and simulating pH of intestinal environment. A Freundlich isotherm was used to describe the adsorption data obtained. The results showed significant decrease of zearalenone concentrations, ranging from 33.5-66.2% per 25 mg of polymer. Adsorption capacity (k) was estimated to be higher than that of previously tested adsorbents, including crospovidone. The data indicate the need to investigate structure peculiarities in order to improve mycotoxin deactivation procedures using PVP derivatives. PMID- 10390852 TI - Accumulation of mercury and its effect on antioxidant enzymes in brain, liver, and kidneys of mice. AB - The effect of mercuric chloride (HgCl2) on the activities of catalase, superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR) and its effect on glutathione (GSH) content were evaluated in different organs (liver, kidneys, and brain) of mice after administration at 0, 0.25, 0.5 and 1.0 mg/kg/day for 14 days. The uptake of mercury shows that the kidneys accumulated the highest levels of mercury compare to brain and liver. The enzyme levels varied in mercury treated organs compare to control. A dose dependent increase of antioxidant enzymes occurred in the liver and kidneys. The increase in enzyme activities correlated with highest mercury accumulation in the kidneys and liver. Mercury is known to generate reactive oxygen species (ROS) in vivo and in vitro, therefore, it is likely that enzyme activities increased to scavenge ROS levels produced as a result of mercury accumulation. Glutathione content increased in liver and kidneys of mercury treated mice compare to control. The results showed that the highest oral dose of mercury significantly increased antioxidant enzymes in kidneys and liver. The increased antioxidant enzymes enhance the antioxidant potential of the organs to reduce oxidative stress. PMID- 10390853 TI - Arsenic toxicity and accumulation in radish as affected by arsenic chemical speciation. AB - Arsenic (As) uptake by Rhapanus sativus L. (radish), cv. Nueva Orleans, growing in soil-less culture conditions was studied in relation to the chemical form and concentration of As. A 4 x 3 factorial experiment was conducted with treatments consisting of four As chemical forms [As(III), As(V), MMAA, DMAA] and three As concentrations (1.0, 2.0, and 5.0 mg As L-1). None of the As treatments were clearly phytotoxic to this radish cultivar. Arsenic phytoavailability was primarily determined by the As chemical form present in the nutrient solution and followed the trend DMAA < or = As(V) < or = As(III) << MMAA. Root and shoot As concentrations significantly increased with increasing As application rates. Monomethyl arsonic acid treatments caused the highest As accumulation in both roots and shoots, and this organic arsenical showed a higher uptake rate than the other As compounds. Inner root As concentrations were, in general, within the normal range for As contents in food crops but root skin As levels were close or above the maximum threshold set for As content in edible fruit, crops and vegetables. The statement that toxicity limits plant As uptake to safe levels was not confirmed in our study. If radish plants are exposed to a large pulse of As, as growth on contaminated nutrient solutions, they may accumulate residues which are unacceptable for animal and human consumption without exhibiting symptoms of phytotoxicity. PMID- 10390854 TI - Accumulation of potentially toxic elements in plants and their transfer to human food chain. AB - Contaminated soils can be a source for crop plants of such elements like As, Cd, Cr, Cu, Ni, Pb, and Zn. The excessive transfer of As, Cu, Ni, and Zn to the food chain is controlled by a "soil-plant barrier"; however, for some elements, including Cd, the soil-plant barrier fails. The level of Cd ingested by average person in USA is about 12 micrograms/day, which is relatively low comparing to Risk Reference Dose (70 micrograms Cd/day) established by USEPA. Food of plant origin is a main source of Cd intake by modern society. Fish and shellfish may be a dominant dietary sources of Hg for some human populations. About half of human Pb intake is through food, of which more than half originates from plants. Dietary intake of Cd and Pb may be increased by application of sludges on cropland with already high levels of these metals. Soils amended with sludges in the USA will be permitted (by USEPA-503 regulations) to accumulate Cr, Cd, Cu, Pb, Hg, Ni, and Se, and Zn to levels from 10 to 100 times the present baseline concentrations. These levels are very permissive by international standards. Because of the limited supply of toxicity data obtained from metals applied in sewage sludge, predictions as to the new regulations will protect crop plants from metal toxicities, and food chain from contamination, are difficult to make. PMID- 10390855 TI - Evaluating peats for their capacities to remove odorous compounds from liquid swine manure using headspace "solid-phase microextraction". AB - This paper reports on research designed to investigate the capacities of different highly characterized peats to remove odorous compounds from liquid swine manure (LSM). Peat types representing a wide range of properties were tested in order to establish which chemical and physical properties might be most indicative of their capacities to remediate odors produced by LSM. Eight percent slurries (of peat/LSM) were measured for odor changes after 24 hours using odor panel and GC/MS-Solid-phase microextraction (GC/MS-SPME) analysis. The GC/MS-SPME and odor panel results indicated that, although all peats tested in this study were found to be effective at removing odor-causing compounds found in LSM, some peats tended to work better than others. Overall, the peats that were the most effective at removing odor-causing compounds tended to have lower bulk densities, ash contents, fulvic acids contents, and guaiacyl lignins contents, and higher water holding capacities, hydraulic conductivities, "total other lignins" contents, hydrogen contents, carbon contents, and total cellulose contents. GC/MS SPME analysis was found to be a reasonably inexpensive and efficient way of conducting this type of research. It allows one to identify a large number of the odor-causing compounds found in LSM, and more importantly, to detect with some precision specific differences in the amounts of these compounds between peat types. PMID- 10390856 TI - Electrospray ionization mass spectrometry of 1-phenyl-3-methyl-5-pyrazolone derivatives of neutral and N-acetylated oligosaccharides. AB - Derivatization using 1-phenyl-3-methyl-5-pyrazolone (PMP) was selected among a number of reported methods for labeling carbohydrates, since it gives a quantitative yield, proceeds through a rapid reaction and involves a simple clean up procedure. Moreover, PMP derivatives provide an increase in sensitivity with ultraviolet and mass spectrometric detection relative to native neutral sugars. Sensitivity studies were carried out using a standard oligosaccharide, tetraglucose. One of the aims of these studies was to determine the minimum amounts of PMP-tetraglucose necessary to generate informative full-scan electrospray ionization (ESI) mass spectra and collision-induced dissociation tandem mass spectra. Another aim was to characterize the fragmentation pattern of PMP derivatives. Quantitative and qualitative studies were also carried out with a typical N-linked oligosaccharide obtained commercially. The PMP-labeled compound underwent directed cleavages which produced fragments containing the reducing end. The native N-linked sugar yielded fragments corresponding to cleavages from both ends of the molecule. Under the same ESI conditions, the N linked oligosaccharide exhibited more lability, or tendency to fragment, than neutral tetraglucose, in both the derivatized and native forms. Also, PMP labeling was shown to enhance sensitivity in the case of a neutral oligosaccharide, i.e. tetraglucose, whereas the labeling of an N-acetylated oligosaccharide, NGA3, did not yield a noticeable improvement in sensitivity. PMID- 10390857 TI - Mass spectrometric analysis of arachidonyl-containing phospholipids in human U937 cells. AB - The human histiocytic lymphoma U937 cell line contains a rich source of the 85 kDa cytosolic phospholipase A2 (cPLA2). DMSO-differentiated U937 cells were used as a model to investigate the free arachidonic acid release, the arachidonate distribution and the phospholipid source of arachidonate upon Ca2+ ionophore stimulation. A combination of several chromatographic and mass spectrometric techniques was employed in this study. The amount of free arachidonic acid (AA) released upon stimulation, the arachidonate content in total lipids and in each of the phospholipid classes were determined by gas chromatography/mass spectrometry (GC/MS). Glycerophosphoethanolamine (GPE) was found to be the major pool of arachidonate in differentiated human U937 cells (55%) and glycerophosphocholine (GPC) and glycerophosphoinositol (GPI) contributed 22 and 8%, respectively. Upon Ca2+ ionophore stimulation, GPE class lost the largest amount of arachidonate, followed by GPC class. GPI class, however, gained a substantial amount of arachidonate. Most of the arachidonate depleted from GPE and GPC was recovered as free AA, some of which was rapidly esterified into GPI species. GC/MS with electron capture negative chemical ionization provided excellent sensitivity for the measurement of arachidonic acid which was derivatized to its pentafluorobenzyl ester. Intact phospholipid molecular species including the arachidonyl-containing phospholipid species were identified using capillary high-performance liquid chromatography/continuous-flow liquid secondary ion mass spectrometry (CF-LSIMS). No specificity was found for releasing free AA among the arachidonyl-containing GPE and GPC species upon Ca2+ ionophore stimulation. CF-LSIMS provided a sensitive and effective means of detecting intact phospholipid species. PMID- 10390858 TI - Identification and quantitation of N-(carboxymethyl)valine adduct in hemoglobin by gas chromatography/mass spectrometry. AB - A sensitive, specific and reproducible method was developed for the quantitation of the hemoglobin (Hb) adduct N-(carboxymethyl)valine (CMV). This adduct is one of various products from the Maillard reaction, involving reducing sugars and amino acids, proteins or other molecules with a free amino group. Such adducts, including N epsilon-(carboxymethyl)lysine (CML), are called advanced glycation end products (AGE) and have been correlated with aging and severity of diabetes in human tissues. This method was developed to examine the CMV-Hb adduct as a possible AGE formed by reaction of Hb with glucose or other oxidation products. CMV was cleaved selectively from isolated globin using pentafluorophenyl isothiocyanate (PFPITC) in a modified Edman degradation at pH 9.5. The carboxyl group of the adduct was derivatized to its methyl ester with diazomethane. The resulting derivative, 5-isopropyl-1-(methyl acetate)-3-pentafluorophenyl-2 thiohydantoin, was detected by gas chromatography/mass spectrometry with selected ion monitoring (GC/SIM/MS). Quantitation was based on the response factor of the derivative molecular ion (m/z 396) from synthesized CMV and N-(2 carboxyethyl)valine (molecular ion m/z 410) as internal standard. This method exhibits reproducibility and linearity in the range 0.2-100 ng CMV. The limit of quantitation (0.2 ng CMV) gave a signal-to-noise ratio greater than 5:1 using a 1:30 sample aliquot. The GC/SIM/MS method can detect CMV adduct in 5 mg globin samples with relative standard deviations less than 5%. This approach avoids tedious acid hydrolysis and interference from other amino acids. The molecular ion and other CMV derivative ion assignments from samples were confirmed by accurate mass determinations using GC/high resolution SIM/MS. Measurements from random mouse, rat and human globin samples gave mean CMV levels of about 6, 5 and 14 nmol g-1 Hb in these species, respectively. PMID- 10390859 TI - Atmospheric pressure ion mobility spectrometry of protonated and sodiated peptides. AB - A number of peptides were studied with electrospray ionization--ion mobility spectrometry/mass spectrometry (ESI-IMS/MS). The ion mobility data were used to calculate the average collision cross sections of the different detected peptide ions in the nitrogen drift gas. By comparing the cross sections of related ions, structural information about the most probable location of the charge and the gas phase ion conformations was deduced. For bradykinin and kemptide, a significant mobility difference between protonated and sodiated species (where sodium replaced a proton in singly and doubly charged peptides) was demonstrated. Surprisingly, the doubly charged sodiated peptides had a smaller collision cross section than the doubly charged protonated ones leading to the conclusion that the gas-phase conformations of these ions are different with respect to intramolecular interactions. PMID- 10390860 TI - Investigation of some covalent and noncovalent complexes by matrix-assisted laser desorption/ionization time-of-flight and electrospray mass spectrometry. AB - Covalent and noncovalent complexes involving bovine superoxide dismutase, bovine alpha-lactalbumin and two variants, A and B, of bovine beta-lactoglobulin have been examined by delayed extraction matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) and electrospray mass spectrometry. Covalent complexation with acrylamide was obtained through the interaction of these proteins with acrylamide monomers while treatment with peroxynitrite yielded complexes with NO2. Some of the complexation sites with acrylamide were reliably identified by performing tryptic digestion followed by MALDI-TOF measurements in the reflection mode. These data demonstrate that the presence of free cysteine in the investigated sequences is not a precondition for the observation of Cys acrylamide complexes. Although the bulk of the present work is dedicated to the characterization of Cys-acrylamide adducts, the preliminary data on noncovalent complexes between two of those proteins and 8-anilinonaphthalene-1-sulfonic acid (ANS) were easily observed under electrospray conditions, while under MALDI-TOF conditions only the complex with beta-lactoglobulin B was clearly evident. The presented data demonstrate the advantage of the parallel use of both ionization techniques for this type of investigation. PMID- 10390861 TI - The identification of unusual bile acid metabolites by tandem mass spectrometry: use of low-energy collision-induced dissociation to produce informative spectra. AB - The use of collision-induced dissociation (CID) tandem mass spectrometry (MS/MS) has been shown to produce fragmentation that is useful for the structural analysis of bile acids and their conjugates. Low-energy CID using a triple quadrupole has been used to help characterise bile acid identity but the majority of work has been conducted using high-energy CID on specialised instrumentation. This paper describes the use of low-energy CID as a rapid method for identification of urinary bile acids and presents some examples of its use in the diagnosis of liver disease in infants. These include the differential diagnosis of peroxisomal disorders, identification of compounds (e.g. 3 beta,7 alpha dihydroxy-5-cholenoic acid 3-sulphate) indicative of 3 beta-hydroxy-delta 5-C27 steroid dehydrogenase/isomerase deficiency and the confirmation of the identity of an unusual bile acid series consisting of different conjugates of lithocholic acid. The use of lithium cationisation and derivatisation with aminosulfonic acids for the analysis of unconjugated and glycine-conjugated bile acids has also been evaluated. PMID- 10390862 TI - A rapid and reliable method for NO quantification and 15NO/14NO determination using isotope ratio mass spectrometry: an application for the detection of NO synthesis in propionibacteria. AB - For the last decade, numerous studies have focused on the positive or toxic effects of nitric oxide (NO) in procaryotic and eucaryotic cells. This gas has fundamental roles in neurotransmission, vasodilatation, cytotoxicity, and intestinal motility. The ability to produce NO by intestinal microflora or probiotic bacteria is unknown. In this preliminary study, we present a rapid and reproducible procedure for NO quantification and 15NO/14NO determination (based on the reaction between nitrite and acidic potassium iodide) by isotope ratio mass spectrometry. Using this method, we have demonstrated for the first time in vitro production of NO by a dietary bacterium (Propionibacterium acidipropionici, Pa 1) under anaerobic culture conditions. Using different sources of nitrogen, we have clearly shown that propionibacteria can synthesize NO from reduction of nitrate or nitrite. In our experimental conditions, NO synthase was not involved in NO production by propionibacteria. PMID- 10390863 TI - Elemental and molecular imaging of human hair using secondary ion mass spectrometry. AB - Secondary ion mass spectrometry (SIMS) is used to image the spatial distribution of elemental and molecular species on the surface and in cross sections of doped human hair using a magnetic sector SIMS instrument operated as an ion microprobe. Analysis of electrically insulating, non-planar hair samples requires one of two different methods of charge compensation to be used depending on the polarity of the sputtered secondary ions. For detection of positive secondary ions, the hair is imaged using a approximately 0.5 micron diameter, 19.5 keV impact energy, O- microbeam with no auxiliary electron bombardment. For detection of negative secondary ions, a approximately 0.2 micron diameter, 14.5 keV impact energy Cs+ microbeam is used in conjunction with normal incidence, low-energy electron bombardment. Both of these methods allow submicrometer spatial resolution elemental and molecular secondary ion images to be obtained from hair samples without metallic coating of the sample surface prior to analysis. Several examples are presented that reflect potential application areas for these analytical methods. PMID- 10390864 TI - Numerical chromosomal abnormalities detected by atomic force microscopy. AB - The numerical abnormalities of human metaphase chromosomes, fixed according to standard procedures for optical microscopy but not treated for banding, were detected by atomic force microscopy (AFM). High-resolution AFM imaging of chromosomes in trisomy 13, 21, and Klinefelter syndrome can be compared directly with the traditional optical image. The unbanded metaphase chromosomes, including the extra ones in trisomic patients showed a structural pattern very similar to G banding. Comparison of AFM images with light microscopic data allows the identification of specific chromosomes, and images of chromosomes showing numerical and structural abnormalities can then be analysed. PMID- 10390865 TI - A structural study of the bovine vaginal fluid at estrus. AB - The present study describes the structural components of the bovine vaginal fluid at estrus by scanning electron microscopy (SEM) following critical point- and freeze-drying preparation procedures. Confocal scanning laser microscopy (CLSM) was also used to evaluate the structural integrity of samples, and a control sample was assessed by adding sperm to the vaginal fluid. Samples were collected from 10 cows at the time of artificial insemination, prepared for SEM by using critical point- and freeze-drying procedures, gold coated, and observed by SEM. Mesh size and filament thickness were measured with an image analyzer. Of the 10 samples processed, 4 were considered altered following critical point drying. Compaction and lack of filaments were observed in these samples. A small area of one sample showed a honey comb-like structure when freeze drying was used. Nonoriented filaments with different thicknesses and with a network-like structure were observed throughout the remainder of the samples. Filaments throughout all samples were also observed by CSLM. After critical point drying, the mesh area ranged from 0.8 to 101.4 microns 2; the minor axis from 0.7 to 10.8 microns; and filament thickness from 40 to 442 nm. Using freeze drying, the mesh area ranged from 0.9 to 493.8 microns 2; the minor axis from 0.7 to 27.5 microns; and filament thickness from 40 to 800 nm. When samples were freeze dried, mesh values were similar to the interstrand channels observed by CSLM. In sperm vaginal fluid samples, following critical point- or freeze-drying procedures, spermatozoa were oriented randomly in the vaginal fluid and did not seem to alter filamentous structure. Our data suggest that the freeze-drying procedure better preserves the true structural dimensions of the vaginal fluid. Furthermore, the filamentous structure of the vaginal fluid does not appear to impede sperm transport. PMID- 10390866 TI - Transformation of Escherichia coli in foodstuffs. AB - The plasmid transfer by transformation of Escherichia coli in 12 foods was investigated under conditions commonly found in processing and storage of food. Transformation occurred in all foods with frequencies of at least 10(-8) when a simplified standard transformation protocol with non-growing cells was applied. Higher rates (ca. 10(-7)) were found in milk, soy drink, tomato and orange juice. Furthermore, E. coli became transformed at temperatures below 5 degrees C, i.e. under conditions highly relevant in storage of perishable foods. In soy drink this condition resulted in frequencies which were even higher than those determined after application of a temperature shift to 37 degrees C. The transformation of cells growing in milk and carrot juice at a constantly kept temperature of 37 degrees C provides evidence for the potential of E. coli to become transformed naturally. With purified DNA frequencies were determined in these substrates of ca. 2.5 x 10(-7) and 2.5 x 10(-8), respectively. Similar frequencies were also obtained in milk containing the crude nucleic acids of homogenised cell suspensions of E. coli (pUC18). Moreover, the release of plasmid DNA from E. coli during food processing and the subsequent uptake of this DNA by growing E. coli cells was shown to take place after homogenisation in milk indicating a horizontal plasmid transfer by transformation of E. coli. PMID- 10390867 TI - Microcystin in cyanobacterial blooms in a Chilean lake. AB - Cyanobacterial blooms dominated by Microcystis sp. occurred in lake Rocuant ("marisma", near Concepcion/Chile) in February 1995 and 1996. In the bloom samples collected in both years the hepatotoxin microcystin was detected by RP HPLC in both samples and in the sample of 1995 also by a toxicity assay using primary rat hepatocytes. In the bloom of 1995, the microcystin content of the dry bloom biomass was determined to be 130 micrograms/g on the basis of the RP-HPLC peak area and 800 micrograms/g on the basis of the rat hepatotoxicity assay, respectively. In the bloom of 1996, RP-HPLC analysis revealed a microcystin content of 8.13 micrograms/g bloom material dry weight. In this year no hepatotoxicity was measured using a concentration range up to 0.8 mg (d. w.) of bloom material per ml in the rat hepatotoxicity assay. This is the first report on the detection of microcystins in Chilean water bodies. PMID- 10390868 TI - Organization of the chromosomal region containing the genes lexA and topA in Thermotoga neapolitana. Primary structure of LexA reveals phylogenetic relevance. AB - The chromosomal region of Thermotoga neapolitana surrounding the gene lexA (4283 bp) was sequenced. In addition to the topoisomerase gene top2A it contained five open reading frames. A part of the cloned region showed high sequence homology with a previously published sequence of Th. maritima and indicated an identical arrangement of genes in both microorganisms. Structural analysis of the LexA protein showed significant, but relatively low overall homology with LexA proteins of other bacteria, especially in the DNA binding region. However, key amino acids for processing and secondary structure elements like the helix-turn helix motif are well conserved. Sequence alignment analysis of the whole protein and the DNA-binding sites of all known LexA sequences uncovers groups of similarity reminding the phylogenetic tree of the Bacteria. A consensus sequence with the SOS- or Cheo-box upstream of the lexA gene of Th. maritima and Th. neapolitana was absent. Together with the phylogenetic distance of the Thermotogales from other bacteria this suggests the presence of a new operator target sequence specific for the Thermotogales, in analogy to the SOS-box for the gamma-group Proteobacteria and the Cheo-box for low- and high-GC Gram-positive bacteria. PMID- 10390869 TI - Specific oligonucleotide probes for in situ detection of a major group of gram positive bacteria with low DNA G + C content. AB - Almost one thousand 16S rRNA sequences of Gram-positive bacteria with a low DNA G + C content from public databases were analyzed using the ARB software package. A signature region was identified between positions 354 and 371 (E. coli numbering) for the Bacillus sub-branch of the Gram-positive bacteria with a low DNA G + C content, the former orders Bacillales and Lactobacillales. Three oligonucleotide probes, namely LGC354A, LGC354B, and LGC354C, were designed to target this diagnostic site. Their fluorescent derivatives were suitable for whole cell detection by fluorescence in situ hybridization (FISH). Hybridization conditions were adjusted for differentiation of target and related non-target reference species. When applying FISH to whole bacterial cells in a sample of activated sludge from a communal wastewater treatment plant, members of the Bacillus sub branch were detected at levels from 0.01% of cells in samples fixed with paraformaldehyde to over 8 percent in the same samples fixed with ethanol and treated with lysozyme. The problems of quantitative in situ analysis of Gram positive bacteria with a low DNA G + C content in biofilm flocs are discussed and recommendations made. Members of the Bacillus sub-branch were detected in different abundances in activated sludge samples from different wastewater plants. PMID- 10390870 TI - Defluvibacter lusatiae gen. nov., sp. nov., a new chlorohenol-degrading member of the alpha-2 subgroup of proteobacteria. AB - The two Gram-negative bacterial strains S1 and S4 were isolated from activated sludge of an industrial waste water treatment plant and exhibited a stable capability to degrade 2,4-dichlorophenol, 4-chloro-2-methylphenol, 4-chlorophenol and phenol. The cells were short rods with a polar flagellum, being mesophilic, strictly aerobic, oxidase-positive, and chemoorganotrophic. They utilized a range of amino acids, but only a restricted number of carbohydrates. Reassociation experiments with DNA from strains S1 and S4 revealed high interstrain similarity, indicating, that both strains belong to the same species. The phylogenetic position was determined by comparison of the almost complete 16S rDNA sequence of strain S1 with sequences of related bacteria. Strain S1 clustered with members of the alpha-2 subgroup of the Proteobacteria by forming a separate lineage within the radiation of Mesorhizobium, Phyllobacterium and Sinorhizobium. Both strains can be differentiated from members of related taxa by a set of physiological and chemotaxonomic properties including the ability to grow with norvaline, L tryptophan, putrescine, glutarate and malonate, and by the presence of spermidine as major polyamine and of 12:0 3OH as fatty acid. Strain S1 is described as type strain of a new species and assigned to a new genus with the proposed name Defluvibacter lusatiae. PMID- 10390871 TI - Genotypic diversity of Acidovorax strains isolated from activated sludge and description of Acidovorax defluvii sp. nov. AB - Fluorescence in situ hybridization of activated sludge samples from a municipal wastewater treatment plant using oligonucleotide probes specific for Acidovorax demonstrated that these bacteria are highly abundant in this environment. For the targeted cultivation of representatives belonging to this genus, isolates grown on agar plates after serial dilution were screened by whole-cell hybridization with specific probes. The obtained strains clustered in two phylogenetic groups as determined by 16S rRNA gene sequence analyses. The isolates of one cluster were phylogenetically and genotypically closely related to A. delafieldii. In contrast, the strains of the other cluster were genotypically and phenotypically distinct from the hitherto known Acidovorax species. Therefore, a new species, Acidovorax defluvii sp. nov., was proposed for these strains. The main characteristics of the newly defined species are as follows: Gram-negative, motile or non-motile rods with rounded ends, often with large polyhydroxybutyrate granules. In broth cultures flocs are formed. Test for cytochrome oxidase is positive with all strains. The majority of strains is catalase positive and reduces nitrate. All strains are metabolically inactive against most carbohydrates and organic acids. Fatty acid patterns are typical for the genus Acidovorax. The guanine-plus-cytosine content of DNAs varies between 62 and 64 mol%. The type strain of A. defluvii is BSB411T (DSM 12644). A new 16S rRNA targeted oligonucleotide probe reacting by in situ hybridization with all known Acidovorax species, including A. defluvii sp. nov., was designed. PMID- 10390873 TI - Identification of Staphylococcus carnosus and Staphylococcus warneri isolated from meat by fluorescent in situ hybridization with 16S rRNA-targeted oligonucleotide probes. AB - 16S rRNA targeted oligonucleotide probes were designed by sequence analysis of an rRNA database to discriminate S. carnosus, S. warneri, and S. saprophyticus species. After establishing hybridization conditions by RNA dot blot hybridization with reference species, our probes were shown to be specific. By in situ hybridization only S-S-S.carno-0440-a-A-23 and S-S-S.war-0180-a-A-23 can specifically detect S. carnosus and S. warneri, respectively. The detection of old cells of S. carnosus 833 was more limited by the permeabilisation than by the low rRNA content. One day old cells could be permeabilized with lysostaphin, whereas young cells were permeabilized with lysozyme. PMID- 10390872 TI - The rice inoculant strain Alcaligenes faecalis A15 is a nitrogen-fixing Pseudomonas stutzeri. AB - The taxonomic position of the nitrogen-fixing rice isolate A15, previously classified as Alcaligenes faecalis, was reinvestigated. On the basis of its small subunit ribosomal RNA (16S rRNA) sequence this strain identifies as Pseudomonas stutzeri. Phenotyping and fatty acid profiling confirm this result. DNA:DNA hybridisations, using the optical renaturation rate method, between strain A15 and Pseudomonas stutzeri LMG 11199T revealed a mean DNA-binding of 77%. The identification was further corroborated by comparative sequence analysis of the oprF gene, which encodes the major outer membrane protein of rRNA homology group I pseudomonads. Furthermore we determined the nifH sequence of this strain and of two putative diazotrophic Pseudomonas spp. and made a comparative analysis with sequences of other diazotrophs. These Pseudomonas NifH sequences cluster with NifH sequences isolated from the rice rhizosphere by PCR and of proteobacteria from the beta and gamma subclasses. PMID- 10390874 TI - Four new species in Lipomyces. AB - For new species in Lipomyces are described. In terms of nuclear genome comparison the genus Lipomyces comprises three species-clusters. A key to the nine species now assigned to the genus, is given. The delimitation of species by (i) rRNA nucleotide sequence analyses of the 18S and 25S subunits and (II) nDNA homology determinations, do not invariably lead to the recognition of congruent taxa. The family of the Lipomycetaceae, nevertheless provides an illuminative model for phylogenetic studies in terms of more appropriate ribosomal nucleotide sequence analyses of both its teleomorphic and anamorphic members. PMID- 10390875 TI - The response of the microbial community of marine sediments to organic carbon input under anaerobic conditions. AB - Cyanobacterial biomass was added to anaerobic sediment to simulate the natural input of complex organic substrate that occurs in nature after algae blooms. Sediments were incubated at 0 degree C, 8 degrees C and 24 degrees C for 13 days. Community dynamics were measured by fluorescence in situ hybridisation (FISH), denaturing gradient gel electrophoresis (DGGE), and sequencing of 16S rDNA PCR products. Metabolic changes were followed by the analysis of total carbon mineralisation, sulfate reduction, and ammonium production rates. The addition of organic material resulted in significant changes in the composition of the microbial community at all temperatures tested. Sulfate reduction was the main mineralisation process detected. However, not sulfate-reducers but rather members of the Cytophaga-Flavobacterium phylogenetic cluster showed the highest increase in the bacterial cells as detected by FISH. We conclude that these organisms play an important role in the anaerobic decomposition of complex organic material perhaps because they are the main catalysts of macromolecule hydrolysis and fermentation. The molecular methods also indicated a stimulation of ribosome synthesis. The detection of a large number of rRNA-rich cells belonging to the Cytophaga-Flavobacterium phylogenetic cluster further supports the importance of their role in the degradation of complex organic material in anaerobic marine sediments. Their detection in high numbers in the field may indicate recent deposition events. PMID- 10390876 TI - Rapid isolation of single microbial cells from mixed natural and laboratory populations with the aid of a micromanipulator. AB - In order to facilitate the isolation of pure cultures from natural habitats we have developed a method for the isolation of single microbial cell clones from a mixed population, e.g. the flora of the termite gut, with the aid of a modern micromanipulator. The separated single prokaryotic or eukaryotic cells were grown after transfer in culture media or they were used for single cell PCR. The micromanipulator was also applied for the removal of nuclei from protozoa, of which the SSU rDNA was directly amplified. PMID- 10390877 TI - Amplified rDNA restriction analysis and further genotypic characterisation of metal-resistant soil bacteria and related facultative hydrogenotrophs. AB - The level of genotypic relationship between czc+ soil bacteria mainly resistant to zinc (but also to various other metals), and related facultative hydrogenotrophs previously assigned to the genera Alcaligenes, Ralstonia, and Burkholderia was evaluated using ARDRA (Amplified Ribosomal DNA Restriction Analysis). The analysis included 44 strains isolated from harsh industrial environments in sediments, soils and wastes with high content of heavy metals. These strains were selected by their ability to grow in the presence of high concentrations of multiple heavy metals and to hybridise with czc or ncc probes. The czc operon confers resistance to cadmium, zinc and cobalt in strain Ralstonia eutropha CH34. The ncc operon confers resistance to nickel, cobalt and cadmium in strain 31A known as Alcaligenes xylosoxidans. The analysis showed a close phylogenetic clustering of the czc+ strains inside the Ralstonia genus despite of their different origins and that the Ralstonia genus contained also the hydrogenotrophs and some catabolic strains assigned to the genus Ralstonia eutropha, strains up to now registrated as CDC IV c-2 strains as well as reference strains belonging to Ralstonia solanacearum and Ralstonia pickettii. The ncc+ strains are phylogenetically less related to each other compared to the czc+ strains. This suggests that the tested czc+ strains and some of the ncc+ strains may be considered as belonging to the genus Ralstonia. Inside this major Ralstonia cluster, a subcluster gathers most of the czc+ isolates maybe giving a clue to define a new species. Besides, from 30 tested strains, 15 metal resistant strains of this subcluster proved to display the unusual mutator phenotype characteristic of the representative strain CH34. PMID- 10390878 TI - Sulfate-reducing bacteria in rice field soil and on rice roots. AB - Rice plants that were grown in flooded rice soil microcosms were examined for their ability to exhibit sulfate reducing activity. Washed excised rice roots showed sulfate reduction potential when incubated in anaerobic medium indicating the presence of sulfate-reducing bacteria. Rice plants, that were incubated in a double-chamber (phylloshpere and rhizosphere separated), showed potential sulfate reduction rates in the anoxic rhizosphere compartment. These rates decreased when oxygen was allowed to penetrate through the aerenchyma system of the plants into the anoxic root compartment, indicating that sulfate reducers on the roots were partially inhibited by oxygen or that sulfate was regenerated by oxidation of reduced S-compounds. The potential activity of sulfate reducers on rice roots was consistent with MPN enumerations showing that H2-utilizing sulfate-reducing bacteria were present in high numbers on the rhizoplane (4.1 x 10(7) g-1 root fresh weight) and in the adjacent rhizosperic soil (2.5 x 10(7) g-1 soil dry weight). Acetate-oxidizing sulfate reducers, on the other hand, showed highest numbers in the unplanted bulk soil (1.9 x 10(6) g-1 soil dry weight). Two sulfate reducing bacteria were isolated from the highest dilutions of the MPN series and were characterized physiologically and phylogenetically. Strain F1-7b which was isolated from the rhizoplane with H2 as electron donor was related to subgroup II of the family Desulfovibrionaceae. Strain EZ-2C2, isolated from the rhizoplane on acetate, grouped together with Desulforhabdus sp. and Syntrophobacter wolinii. Other strains of sulfate-reducing bacteria originated from bulk soil of rice soil microcosms and were isolated using different electron donors. From these isolates, strains R-AcA1, R-IbutA1, R-PimA1 and R-AcetonA170 were Gram-positive bacteria which were affiliated with the genus Desulfotomaculum. The other isolates were members of subgroup II of the Desulfovibrionaceae (R-SucA1 and R LacA1), were related to Desulforhabdus sp. (strain BKA11), Desulfobulbus (R PropA1), or culstered between Desulfobotulus sapovorans and Desulfosarcina variabilis (R-ButA1 and R-CaprA1). PMID- 10390879 TI - Phylogeny, ribosomal RNA gene typing and relative abundance of new Pseudomonas species (sensu stricto) isolated from two pinyon-juniper woodland soils of the arid southwest U.S. AB - Rhizosphere-inhabiting Pseudomonas species interact with plant roots and may be important for plant performance under stressful environmental conditions. A comparison was conducted of culturable Pseudomonas isolates associated with pinyon rhizosphere and between-tree interspace areas in a hot, dry, volcanic cinder field and an adjacent sandy loam soil, in order to identify Pseudomonas species which may be involved in pinyon pine survival under stressful conditions. From a collection of 800 isolates, eleven isolates exhibiting different colony morphology were selected for 16S ribosomal RNA gene sequencing. Phylogenetic analysis of rDNA sequences from the eleven field isolates, forty-six described Pseudomonas species, and thirty-four previously characterized environmental isolates indicated that the isolates from the cinders and sandy loam soil clustered into three groups. The field isolates were distinct from any of the named species or other environmental isolates. Oligonucleotide primer pairs that differentiated three field isolate groups were designed from the 16S rDNA sequences, and eight hundred Pseudomonas field isolates cultured from pinyon rhizospheres and interspaces in the cinders and sandy loam soils were typed into the three groups using PCR assays. The composition of Pseudomonas populations in four environments was significantly different. The relative abundance of the three rDNA-based groups appeared to be affected by both the soil type and the pinyon rhizosphere. PMID- 10390880 TI - [Clinical application of soluble transferrin receptor]. PMID- 10390881 TI - [Current trends in the diagnosis of disseminated intravascular coagulation in Japan: findings of questionnaire survey]. PMID- 10390882 TI - [An approach to the problems posed by diagnostic criteria for disseminated intravascular coagulation]. PMID- 10390883 TI - [Tissue factor pathway in disseminated intravascular coagulation]. PMID- 10390884 TI - [Role of endothelial cell dysfunction in disseminated intravascular coagulation]. PMID- 10390885 TI - [Control of thrombosis by retinoids and vitamin D3 derivatives]. PMID- 10390887 TI - [Hematologic improvement of Pearson's syndrome confirmed by mitochondrial DNA analysis]. AB - We report on an 8-month-old girl with Pearson's syndrome who presented with transfusion-dependent pancytopenia, exocrine pancreatic dysfunction, and lactic acidosis. Bone marrow findings were consistent with sideroblastic anemia and marked vacuolization of myeloid and erythroid precursors. Southern blot analysis of mitochondrial DNA (mtDNA) revealed a 4.5 kb deletion in peripheral blood cells. Gradual hematologic improvement was observed thereafter, and the patient was relieved of the need for blood transfusions. We were able to confirm a decrease of the mtDNA deletion in lymphocytes as well as in lymphoblastoid cell lines cultured from peripheral lymphocytes as the patient made steady hematologic progress. PMID- 10390886 TI - [Clinical trial of antithymocyte globulin for prophylaxis of acute graft-versus host disease in pediatric recipients of bone marrow transplantation from unrelated donors]. AB - We studied the effectiveness of antithymocyte globulin (ATG) in preventing acute graft-versus-host disease (a-GVHD) in children who received bone marrow transplants from unrelated HLA-matched donors at one institution. Of 39 patients who received transplants between 1993 and 1997, 23 were given ATG on the basis of informed consent. Either Thymoglobulin (Pasteur Merieux, 2.5 mg/kg/day) or Lymphoglobulin (15 mg/kg/day) was administered for 4 days. a-GVHD (> or = grade II) developed in 33% of the ATG group (n = 21) and in 44% of the non-ATG group (n = 16). Although a-GVHD (> or = grade II) appeared less frequent in the ATG group, the difference was not statistically significant. Among the subjects with hematological malignancies, no significant difference was observed in frequency of a-GVHD (> or = grade II) or 3-year survival rate for the ATG group (n = 10) and non-ATG group (n = 16). However, the incidence of cytomegalovirus infection was much higher (p < 0.01) in the ATG group (70%) than in the non-ATG group (19%). From this study, we were not able to confirm the benefits of ATG as described by other investigators. PMID- 10390888 TI - [Hyperhomocysteinemia: development of deep vein thrombosis in another location during heparin anticoagulation therapy]. AB - A 45-year-old man was admitted complaining of chest pain and pain and edema in the left lower extremity. Ultrasonography and venography results yielded a diagnosis of left femoral vein thrombosis, and pulmonary embolism was diagnosed later. Intravenous heparin therapy (10,000 IU/day) improved the patient's clinical signs. During this therapy, however, pain and edema of the right lower extremity developed, leading to a diagnosis of right femoral vein thrombosis. The patient was admitted to our hospital. At that time, coagulation studies showed an FDP level of 44.7 micrograms/ml and an FDP-DD level of 24.5 micrograms/ml. We surmised that the bilateral deep vein thrombosis had been caused by hyperhomocysteinemia (17.8 mumol/l). Genetic and other acquired risk factors for thrombophilia were ruled out. The patient's clinical signs again improved as a result of intravenous heparin therapy (15,000 IU/day), and FDP and FDP-DD levels returned to normal. We concluded that hyperhomocysteinemia is a risk factor for thrombosis and that it can generate thrombosis in other locations even during heparin therapy. PMID- 10390889 TI - [Epstein-Barr virus associated Richter's syndrome accompanied by interstitial pneumonia]. AB - A 65-year-old man who had an 8-year history of chronic lymphocytic leukemia was admitted to our hospital on February 19, 1998 because of high fever, dry cough, and weight loss. Laboratory data on admission included serum lactate dehydrogenase at 980 IU/l, CRP at 21.8 mg/dl, and soluble interleukin-2 receptor at 7,280 U/ml. The results of serological tests for Epstein-Barr virus (EBV) antibodies were as follows: EBV capsid antigen IgG 1:2560, EBV early antigen IgG 1:640, and EBV nuclear antigens 1:20. Computed tomography revealed diffuse interstitial pneumonia in both lungs, hepatosplenomegaly with multiple nodules, and enlarged intra-abdominal lymph nodes. In addition, Gallium-67 scintigraphy demonstrated abnormal accumulations. Although the patient initially responded well to combination chemotherapy, he eventually deteriorated and died on November 2, 1988, despite salvage chemotherapy. Postmortem needle biopsy specimens from the liver and spleen revealed diffuse proliferation of polymorphic large lymphoma cells. The lymphoma cells were positive for L-26, latent membrane protein 1, and EBV nuclear antigen, but negative for UCHL-1 and CD3, 5, 10, and 30. In situ hybridization procedures disclosed the presence of EBV-encoded small RNA in lymphoma cells. These findings suggested the possibility of association with EBV infection in some cases of Richter's syndrome. PMID- 10390890 TI - [Marked hematologic improvement despite deterioration of marrow cell dysplasia in a refractory anemia patient treated with vitamin D3]. AB - A 69-year-old man was admitted to our hospital due to pancytopenia and a marked bleeding tendency. On admission, he had a white cell count of 2.8 x 10(9)/l, hemoglobin level of 6.0 g/dl, and a platelet count of 3 x 10(9)/l. He was given a diagnosis of refractory anemia on the basis of bone marrow aspiration findings, which disclosed trilineage myelodysplasia. After discharge, the patient remained dependent on blood transfusions. The sole administration of an active form of vitamin D3 (calcitriol) was started in July 1997, and one and a half years later, the patient's transfusion dependency disappeared. However, bone marrow aspiration findings at this point disclosed marked cell dysplasia of erythroid lineage and a prognostically unfavorable chromosomal abnormality (monosomy 7) that had not been found during the initial examination. Nonetheless, the patient's hemoglobin level and platelet count increased to more than 9 g/dl and about 1.0 x 10(11)/l, respectively. The finding in this case suggested that vitamin D3 therapy is useful for refractory anemia even if aggravated marrow cell dysplasia and cytogenetic anomalies develop. PMID- 10390891 TI - [Acute vincristine neurotoxicity in a non-Hodgkin's lymphoma patient with Charcot Marie-Tooth disease]. AB - A 44-year-old, previously healthy man with a diagnosis of non-Hodgkin's lymphoma (NHL, diffuse large B-cell type, stage IIA) was treated with combination chemotherapy including vincristine (VCR). After receiving a cumulative dose of VCR, he experienced rapid and marked weakening which progressed to quadriplegia and bulbar palsy. Prior to this therapy, the patient had no neurological problems, and his siblings were asymptomatic. Physical examination identified pes cavus (hollow foot), and electrodiagnostic studies showed markedly slower nerve conduction velocity of myelinated fibers, with abundant "onion bulb" formations. Chromosomal analysis detected 17p11.2-12 duplication, thus yielding a diagnosis of Charcot-Marie-Tooth (CMT) 1A. CMT disease is a familial neuromuscular disorder, and the incidence is approximately 1 in 2,500. We concluded that if CMT disease is diagnosed, vincristine should be avoided due to the potential severity of neurotoxicity to small doses. PMID- 10390892 TI - [Hypereosinophilic syndrome complicated by myelofibrosis]. AB - Hypereosinophilic syndrome (HES) with myelofibrosis was diagnosed in a 36-year old man on the basis of bone marrow biopsy findings and clinical features. Although the patient was treated with steroid (1 mg/kg), hydroxyurea, and immunosuppressive therapy, eosinophilia persisted. Patients with HES and myelofibrosis are usually unresponsive to antineoplastic agents and/or immunosuppressants. However, cyclosporin may be an effective alternative for such patients. PMID- 10390893 TI - [Autopsy case of intravascular lymphomatosis with pancreatic carcinoma]. AB - We report an autopsy case of intravascular lymphomatosis (IVL) that arose after radiation therapy and chemotherapy for an inoperable pancreatic carcinoma. A 66 year-old man who suffered from diabetes mellitus and pancreatic carcinoma presented with aggressive progression of consciousness disturbance and high fever. The laboratory findings disclosed marked thrombocytopenia, hypercalcemia, and elevated serum PTH-related peptide. The patient soon died of ventricular fibrillation due to uncontrollable hypercalcemia. Postmortem examination with immunohistochemical analysis revealed a well-differentiated tubular adenocarcinoma in the pancreatic body as well as an accumulation of neoplastic B lymphocytes in small vessels throughout the body without systemic lymphadenopathy. To our knowledge, double neoplasms including IVL are extremely rare. PMID- 10390894 TI - [Development of cerebellar astrocytoma in a patient with acute myeloid leukemia 8 years after his second bone marrow transplant]. AB - We report a case of cerebellar astrocytoma occurring 8 years after the second bone marrow transplantation (BMT) in 32-year-old man. The patient was admitted to our hospital in December 1997 because of dysarthria and gait disturbance. He had been treated earlier for acute myeloid leukemia (AML M2) with chemotherapy and cranial irradiation followed by allogeneic BMT from a sibling in december 1988. Three months after the first BMT, testicular relapse was observed and followed by systemic relapse. The patient received reinduction therapy and a second successful BMT. He had been well until about 1 month before admission to our hospital. Neurological examination revealed left cerebellar ataxia, and brain magnetic resonance imaging disclosed a left cerebellar tumor. The tumor was surgically resected and a histological diagnosis of cerebellar astrocytoma was made. The patient was further treated by irradiation for residual tumor and discharged without progression of the disease. PMID- 10390895 TI - [Asthma management by peak expiratory flow]. PMID- 10390896 TI - [Roles of eosinophils in allergic skin diseases]. PMID- 10390897 TI - [Degranulation of human eosinophils in nasal allergy]. AB - The localization of eosinophil peroxidase (EPO), one of the basic proteins in eosinophil grunules, was studied in the nasal mucosae of nasal allergy patients. EPO in granules was initially induced into the cytoplasm via finetubules and then diffused into the extracellular space through micro-destruction of the plasma membrane. When the eosinophil lysis was advanced a large amount of granule countents was released into the extracellular space. The total number of eosinophils which migrated to epithelial region was not significantly different between in the damaged epithelium and in the intact one. But the rate of cytolytic eosinophils was significantly higher in the area of severely shedded epithelium than in the area of intact epithelium. In conclusion, the degranulation of eosinopils in nasal allergy was induced by lysis rather than by exocytosis and many cytolytic eosinophils caused epithelial shedding. PMID- 10390898 TI - [A consideration for indexes of global improvement rating by physicians in bronchial asthma]. AB - As the minimum items required for GIR evaluation, morning Peak Respiratory Flow Rate (PEFR), evening PEFR, asthma scores and symptom scores were selected based on Cronbach's alpha, an index of internal consistency. Optimum categorization of total improvement scores (TS), or principal component scores (PCS) was conducted based on agreement with the physician's evaluation of GIR. Good agreement was observed between optimally categorized moderate and marked improvement rate in TS or PCS and moderate and marked improvement rate in the physician's evaluation of GIR. The rate of agreement between optimally categorized TS or PCS and distribution of GIR was about 93% if allowance is made for discrepancy of one stage. Our assessment that TS which is objective and practical was useful as criteria for evaluating GIR. PMID- 10390899 TI - [Change of housing environment and withdrawal of corticosteroid as treatments of atopic dermatitis]. AB - For the purpose of searching for the factors for improvement in cutaneous lesions of atopic dermatitis, we evaluated two factors, namely, change in the housing environment and withdrawal of corticosteroid in a group of patients whose clinical course had been observed for more than one year. The condition of illness was judged by a combination of VAS scoring of cutaneous lesions, laboratory data (IgE, LDH, eosinophil) and evaluation by the patient himself. The results showed a significant difference (p = 0.143) by Fisher's direct probability for the tendency to improve in the group in which the housing environment had changed and p = 0.266 for the tendency to improve in the group in which corticosteroid was withdrawn. When the tendency to improve by either change in the housing environment or corticosteroid withdrawal was examined, the p value was 0.028 at 5% level of significance. That is, the findings in this study suggest that atopic dermatitis will improve in the presence of either or both of these two factors at 5% level of significance. Factors regulating the condition of illness in atopic dermatitis are diverse. Analysis using only one of these factors cannot easily show a statistically significant difference. So studies involving two or more factors are needed. PMID- 10390900 TI - [A trial of new protocol of rush immunotherapy with standardized mite antigen]. AB - The objective of the present study was (1) to establish a new protocol of rush immunotherapy (IT) using a purified mite antigen for the treatment of house-dust mite-sensitive bronchial asthma. Ten adult asthmatics were enrolled in the initial trial which was based on a 7-day protocol using a house-dust antigen, to determine the optimal maintenance dose of mite antigen. No systemic side effects were observed when the antigen dose was lower than 50 AU. Consequently, the duration of rush IT was shortened to 5 days, and the maintenance dose was determined as 50 AU in the new protocol. To confirm its safety, another ten asthmatics were enrolled in a second trial. Rush IT using the new protocol was able to be performed without any systemic side effects in all patients except one who showed urticaria at 40 AU. These results suggest that the 5-day protocol of rush IT using the mite antigen is safe in patients with house-dust-mite-sensitive asthma. PMID- 10390901 TI - [Development and evaluation of parent-related quality of life questionnaires for asthmatic children and their parents or caregivers--evaluation of treatment process with sustained-release theophylline dry syrup]. AB - A quality of life (QOL) questionnaire for asthmatic children and their parents or caregivers was shown to exhibit reliability, factorial validity and possibility for application study in our previous report. The aim of this study was to examine longitudinal study of the questionnaires, including changes of QOL, responsibility to asthmatic symptoms, consistency with conventional asthma scores and clinical outcomes during treatment with theophylline dry syrup for about three months. Overall QOL scores increased significantly (p < 0.01) with improvement in clinical outcomes. Longitudinal changes of QOL were compared with differences (after-before) and changes of ratio [(after-before/before)] in QOL by correlation analysis between differences of asthma scores. Changes of ratio in QOL were reflected by the clinical outcomes. Results of investigation of correlation between differences in asthma scores and changes of ratio in QOL domains were considered as follows. Physical domain was correlated with asthma symptoms, given the moderate correlation (rho = 0.497). Social and family domains showed fair correlation (rho = 0.382 - 0.384 respectively), both domains were hard to correlate with differences in asthma scores. Emotional domain showed fair correlation (rho = 0.264) for reflecting family's emotion. Poor correlation (rho = 0.071) was observed in individual growth domain which was necessary to evaluate for longer term. Changes of ratio in QOL responded significantly (p < 0.01) to improved and worst-unchanged groups which were categorized by the differences in asthma scores. In conclusion, QOL questionnaires showed longitudinal validity for clinical outcomes and responsibility to asthmatic symptoms. It yields useful information for understanding how asthmatic symptoms affect parents or caregivers and investigating what treatments should be prescribed for the patients with asthma. PMID- 10390902 TI - [Degranulation of eosinophils by IgG antibody to Candida antigen]. AB - The pathophysiological role of IgG antibody to fungi antigen widely distributed in environment such as Candida albicans in bronchial asthma has not been clarified. Wells of microtiter plate were coated with the extract of Candida albicans and then IgG antibody was immobilized on the wells by incubation with patient's serum. After cultivation of eosinophils on the well, degranulation of eosinophils, as assessed by quantitation of EPX in the supernatant, has been observed. Degranulation was completely abrogated after depletion of IgG in the serum and also decreased by incubation of the cells with anti-CD32 antibody, or anti-CD18 antibody, but not anti-CD23 antibody. Immune complex, which had been prepared by incubation of the extract of Candida albicans with patient's serum, also evoked degranulation of eosinophils. We have examined whether degranulation can be induced by two purified antigens of Candida albicans, i.e., mannan A and acid protease. IgG antibody to acid protease was detected at no or minimal levels in most sera and the antigen did not induce degranulation. On the other hand, mannan A induced degranulation. This observation may be due to response for the presence of IgG antibody to mannan A in the sera. These results suggest that immobilized IgG induced degranulation of eosinophils through Fc gamma RII (CD 32) on eosinophils and mannan A is a major allergen associated with IgG-induced eosinophil degranulation. PMID- 10390903 TI - [A case of cold urticaria with histamine release by a possible skin intrinsic mechanism]. PMID- 10390904 TI - Characterization and proposed nomenclature of epidemic strains of methicillin resistant Staphylococcus aureus in Canada. AB - We hope that standardized nomenclature for identifying epidemic MRSA strains prevalent in Canadian hospitals will be helpful to physicians and infection control practitioners attempting to understand and control the spread of the organism in health-care facilities. It is anticipated that as MRSA continues to evolve in Canadian health-care facilities other strains may be recognized as "epidemic"; as these strains become better characterized they may be added to those designated above. Laboratory physicians and infection control personnel are invited to submit strains that may warrant characterization and designation as a Canadian epidemic strain to the Laboratory Centre for Disease Control, Health Canada, Winnipeg, Manitoba. PMID- 10390907 TI - AIDS and HIV infection in the United Kingdom: monthly report. PMID- 10390905 TI - Outbreak of Hendra-like virus--Malaysia and Singapore, 1998-1999. PMID- 10390909 TI - Spiral CT and pulmonary embolism: is the emperor still unclothed? PMID- 10390908 TI - Success in using non-invasive mechanical ventilation is predicted by patient pathophysiology. A retrospective review of 199 patients. AB - BACKGROUND: To determine the effect of patient pathophysiology on the success or failure of noninvasive mechanical ventilation as determined by the need for subsequent endotracheal intubation. METHODS: Center-based, retrospective case analysis of all patients placed on non-invasive mechanical ventilation for acute respiratory distress. MEASUREMENTS AND RESULTS: Retrospective chart review was performed on patients who were treated with non-invasive mechanical ventilation from 1/94-6/97. Patients were divided into those with rapidly reversible disease processes (RRDP), and those with non rapidly reversible disease processes (NRRP). The proportion of patients requiring subsequent intubation in each group was compared. There were 116 patients with RRDP and 83 patients with NRRD. In the RRDP group, 85.8 percent (95 percent confidence interval 80.9-90.7 percent) of patients did not require intubation. In the NRRP group, 35.7 percent (95 percent C.I. 29.0-42.4 percent) did not require intubation (X2 analysis, p < .001). Multivariate analysis identified patient pathophysiology as the only variable associated with subsequent intubation. CONCLUSIONS: Patient pathophysiology based upon their expected clinical course can be used to predict the success of non invasive mechanical ventilation (NIMV). Patients with NRRD, such as pneumonia, myocardial infarct and sepsis, are much more likely to fail non-invasive mechanical ventilation and require subsequent endotracheal intubation. PMID- 10390910 TI - 1998 laboratory satisfaction survey. Conducted by the Medical Society of Delaware. PMID- 10390911 TI - Gun control as a public health issue. PMID- 10390912 TI - [The role of endothelium in normal pregnancy and pregnancy complicated by preeclampsia]. AB - Endothelial cell dysfunction is thought to play a role in preeclampsia and the reduced production by vascular endothelial cells of the antiaggregatory and vasodilatory factors is well documented. The present study was designed to evaluate endothelial cells function in preeclamptic and healthy pregnant subjects. The nitric oxide plasma concentration in women with preeclampsia was significantly lower as compared with normotensive pregnant women. A significant increase in ET concentration was found in preeclamptic women as compared with normal pregnant patients and normal non-pregnant. The plasma concentrations of von Willebrand factor were significantly increased in healthy pregnancy as compared with preeclamptic patients. The results of our study demonstrate a significant endothelial cells damage in preeclamptic patients. Whether these observations contribute to the vascular pathophysiologic features of preeclampsia remains to be proved. PMID- 10390913 TI - [The role of nitric oxide and blood platelets in pathogenesis of preeclampsia]. AB - The aim of this study was to evaluate the concentration of nitric oxide and the platelet function in preeclamptic and normal pregnant women. The patients with preeclampsia had new hypertension (diastolic blood pressure consistently > or = 90 mmHg with previously lower readings), new proteinuria and generalized oedema that subsequently regressed after delivery. Blood was collected by routine forearm venipuncture before delivery. The following parameters were evaluated: nitric oxide, beta-TG and PF4. The nitric oxide plasma concentration in women with preeclampsia was significantly lower compared with normotensive pregnant women. beta-TG and PF4 concentrations were significantly increased in patients with preeclampsia. Whether these observations contribute to the vascular pathophysiologic features of preeclampsia remains to be proved. PMID- 10390914 TI - [Evaluation of the effectiveness of a low-dose aspirin in the treatment of intrauterine growth retardation (IUGR)]. AB - OBJECTIVE: To evaluate the benefits of IUGR treatment by low doses of acetylsalicylic acid (ASA) (1.5 mg/kg) compared to the standard method. The study was based on the reports that aspirin at low doses shifts prostacyclin/tromboxan A2 balance to the dominance of prostacyclin by inhibiting cyclooxygenase activity in platelets, which results in the improvement of the utero-placental circulation. MATERIAL AND METHOD: 31 pregnant women with diagnosed fetal IUGR were randomly assigned to two groups, receiving either low-dose ASA (n = 22) or the standard treatment (Sadamin, Partusisten, glucose i.v., amino acids i.v.) for 10 days. Ultrasound examination of the biometric parameters of the fetus (BPD, AC, FL) was performed and estimated fetal weight (EFW) calculated before and after treatment. The birthweight of infants in the two examined groups was compared. RESULTS: The mean increase in EFW was higher in the aspirin-treated group compared to that receiving standard treatment (478 g vs 246 g, p < 0.05). In all the biometric parameters under study a higher increase was noted in the group with aspirin treatment; however, the difference was not statistically significant. The mean birthweight was found to be higher in the ASA group as well (2856 g vs 2511 g). The frequency of small-for-gestational-age (SGA) infants (birth weight below 10th percentile) was lower in the ASA group than in the controls (27% vs 55%). The low-dose aspirin therapy did not produce any adverse side-effects either among mothers of infants. CONCLUSION: The treatment with low doses of aspirin reduces the proportion of SGA babies and increases birthweight in the case of a diagnosed fetal growth retardation. Since the number of subjects in this study was relatively small, further clinical trials are necessary to evaluate the effectiveness of IUGR treatment by low-dose aspirin. PMID- 10390915 TI - [Malonyldialdehyde and total antioxidant status in the peritoneal fluid of infertile women]. AB - OBJECTIVE: To estimate the concentration of malonyldialdehyde (MDA) and total antioxidant status in the peritoneal fluid (PF) of patients with unexplained infertility (UI) and infertile women with minimal and mild endometriosis. MATERIALS AND METHODS: PF was obtained during laparoscopy from 8 women with UI, 12 infertile women with endometriosis (I degree and II degrees rAFS) and 10 women with benign noninflammatory ovarian tumours. All laparoscopies were performed in the follicular phase of the cycle. MDA concentration was measured according to Ledwozyw method, TAS was measured spectrophotometrically using RANDOX diagnostic reagent system. RESULTS: We found significantly higher concentration of MDA in PF from both patients with UI (p = 0.03) and with endometriosis (p = 0.046) compared to the control group. TAS was significantly (p = 0.027) higher in PF of women with UI but did not differ significantly (p = 0.49) between patients with endometriosis and controls. CONCLUSIONS: Our results show that an imbalance between lipid peroxides and the antioxidant system in PF environment may be one of the main factors responsible for the UI. In the group with endometriosis a marginally significant difference in MDA levels, no significant differences in TAS and data from the literature, suggest that accelerated lipid peroxidation in PF doesn't appear to play a role in the endometriosis associated infertility. PMID- 10390916 TI - [Results of surgical treatment of urinary stress incontinence women]. AB - OBJECTIVES: Analysis of early and late effects of surgical treatment at women suffering from the urinary stress incontinence. MATERIALS AND METHODS: There were examined 52 patients treated surgically with urinary stress incontinence, and treated with perineoplasty (group I), treated with Marshall-Marchetti-Krantz procedure (group II), or both types of operations at the same time (group III). Early results of treatment were estimated at the 8th-10th day after surgical procedure, and late after two-eight years considering clinical and ultrasound examinations, and individual feelings of the patient. RESULTS: A very good therapeutical effect of surgical treatment was obtained at 75% of patients, but the late one at 42% of patients. The early effect was the best within the group II (88%), whereas the late one within the group I, the worst results were obtained within the group III. The period between the procedure and repeated disorder was the longest within the group I. At women operated before menopause there were noted very good effects at 61% of them, but at those patients operated after menopause--at 38%, whereas the late repeated disorder often concerned the women operated before than after menopause (adequately 65 and 35%). CONCLUSION: The treatment of the urinary stress incontinence employing perineoplasty compared with Marshall-Marchetti-Krantz procedure proved slighter risk of recurrence, slighter intensity and longer period without disorders. The application of both perineoplasty and Marshall-Marchetti-Krantz procedures simultaneously are more ineffective than of those procedures applied individually. Early effects of surgical treatment of urinary stress incontinence are better at patients operated before menopause, however more stable effect was noted at women operated after menopause. PMID- 10390917 TI - [Borderline ovarian tumors: clinical analysis of 114 cases]. AB - OBJECTIVES: The aim of this study was to analyse the group of patients with borderline ovarian tumours. DESIGN: The analysis included 114 patients, operated for ovarian tumours of borderline malignancy in the Gynaecological Department of Medical University of Gdansk between 1978-1997. The study takes into account comparison of: age of patients, obstetrical past, clinical signs and symptoms, clinical stage (according to FIGO), type of surgery, tumour pathology, post surgical treatment. Furthermore, long-term follow up was assessed. RESULTS: Middle age in the group was 48 years, main symptoms: pain of lower part of abdomen (47%) and ascites (26%). 92% of tumours were recognised in stage I (FIGO). 44.7% of the tumours were histological serous, 36% were mucinous. All patients were treated by surgery and 12% received additional treatment. Mean follow up was 104 months (1-247). 9.6% of the patients died because of main disease, next ten persons for reasons not connected with the main disease. 5 year survival rate was 91.2%, 10-year 84.2%. CONCLUSIONS: 1. Borderline ovarian tumours are most frequently recognised in stage I. 2. Serous and mucinous borderline ovarian are dominant. 3. Type of surgery is dependent on age of patients, obstetrical past, clinical stage and tumour pathology. 4. Prognosis in borderline ovarian tumours is excellent. PMID- 10390918 TI - [A case of cystic adenomatoid lung malformation treated by thoracocentesis]. AB - A case of Cystic Adenomatoid Lung Malformation (CALM) treated by serial thoracocenteses followed by implantation of the feto-amniotic shunt is presented. The idea was to prevent prolonged lung compression leading to lung growth retardation and maturation restriction. Postnatally the baby presented good respiratory condition. She underwent lung lobectomy in the 2nd day of life. The histological assessment confirmed the ultrasound diagnosis of CALM. The postoperative period was not complicated. The method of feto-amniotic shunting is presented as promising in the treatment of CALM prenatally. PMID- 10390919 TI - [The deterioration of the parameters of the human semen: myth or reality?]. AB - Numerous publications in the world's literature on the base of multiply retrospective analyses suggest a population-wide decline in the quality of human semen over the past 50 years. The average sperm density, the mean sperm volume and the percentage of the typical spermatozoids appear to have decreased. Furthermore, a significant increase in the incidence of both genitourinary abnormalities (hypospadiasis, cryptorchidism) and testicular cancers in the general population has been reported. The data vary among different geographic regions. The possible adverse effect of the environmental pollutants with oestrogenic activity on male reproductive ability has been considered. These chemicals can affect the foetal development of male gonads and also impair testes during maturity, which finally results in disturbances in testicular functions and leads to the reduction of male fertility. PMID- 10390920 TI - [Usefulness of selected circulating biochemical markers in diagnosis and monitoring of endometriosis]. AB - A review of the literature for searching of biochemical marker of endometriosis is presented. The investigations with CA 125, antiendometrial antibodies, placental protein 14 and others have not established a sensitive screening test for predicting endometriosis, especially in early stages of the disease. Most authors confirm usefulness of those markers in monitoring therapy and predicting recurrences. This may allow to avoid commonly performed "second look" laparoscopy. PMID- 10390921 TI - [Leiomyomatous neoplasms of the esophagus]. PMID- 10390922 TI - [Changes in the organization of the extracellular matrix at the skin level during diabetes]. AB - The ultrastructural study of cutaneous biopsies in diabetic patients highlighted in different phases of disease a progressive alteration of extracellular matrix (E.M.). In the initial phase of disease the morphologic aspect showed an increased accumulation of proteoglycan biglycan, laminin, fibronectin and type IV collagen. These components are responsible for the lamina lucida expansion and are induced by TGF-beta. In the last phase of the disease, an accumulation and a defective organization of E.M. component arises. Type V collagen, normally not present in the skin, is observed. In patients with over ten years of diabetic history, the morphological aspect is defined by a progressive disorganization of E.M. The formation of a vicious circle is responsible for the progressive remodeling of E.M. This process may be linked to the not enzymatic glycosylation of E.M., due to several episodes of hyperglycemia, and to autoinductive mechanisms of TGF-beta. These mechanisms are responsible for the cytokine synthesis and for the E.M. inhibition of degradation. PMID- 10390923 TI - [Primary carcinoma of the gastric stump. Physiopathological, diagnostic and therapeutic considerations]. AB - Even though the primary carcinoma of the gastric stump is a tumor that will diminish in frequency in the years to come, it is still a topic of scientific studies. The authors report their experience with four cases of primary carcinoma of the gastric stump treated surgically as compared to 89 cases of carcinoma of the stomach operated in the same period. After some comments on the etiopathogenesis that is at the basis of the neoplastic mutations of the remaining gastric epithelium, clinical, prognostic and pathologic features that differentiate this type of tumor from those which develop in unoperated stomachs are examined and, then, the most frequent therapeutic approaches are illustrated. In conclusion, it is sustained that patients who have undergone partial gastrectomy for benign disease should be closely followed-up from the tenth year after the operation and, in any case, in those who are over fifty years of age. PMID- 10390924 TI - [Recurrent liver metastases from colorectal cancer: their surgical treatment]. AB - Records of 8 patients undergoing repeated operation for isolated hepatic metastasis were reviewed for operative morbidity and mortality, survival, disease free survival. The mean interval between the initial colon operation and first hepatic resection and that between the first and the second hepatic operation were calculated. Both were found to be correlated with survival of these patients. Repeat hepatic operation for recurrent colorectal metastasis to the liver yields comparable results to first hepatic resections in terms of operative mortality and morbidity, survival, disease-free survival. Repeated hepatic operation is the most successful for of treatment for isolated recurrent colorectal metastasis. PMID- 10390925 TI - [Traditional cholecystectomy in a patient with situs viscerum inversus partialis]. AB - The authors discuss the etiology of situs viscerum inversus partialis (SVIP), the associated anomalies and the malformations and the relative clinical problems, reviewing the international literature. The present a case of a 51 years old female with a diagnosis of umbilical hernia and cystocele. During hospitalization the patient complained a typical acute cholecystitis pain. The patient underwent ultrasonography, CT scan, and MNR that allowed us to diagnosis a calculous cholecystitis with SVIP and was elected for an open cholecystectomy. The Authors describe the surgical technique in relation to the anatomic anomalies, in particular vascular ones, that were discovered with the imaging studies and confirmed at laparotomy. PMID- 10390926 TI - [Neoplastic obstruction of the vena cava inferior in general surgery]. AB - Patients with primary or secondary tumoral occlusion of the inferior vena cava are difficult to be managed with safety and success. Nevertheless, their survival may be prolonged by an aggressive surgical approach according to the technical advances of liver transplantation. In fact, it is possible to perform a tumoral exeresis including the inferior vena cava by a total vascular exclusion of the liver (HVE) and a pump-driven veno-venous bypass (ECC). The Authors report the management of 8 patients with inferior caval tumoral involvement (8 M, 1 F, mean age 63.7 yrs). Vascular occlusion was caused by caval leiomyosarcoma (n 1), renal cell carcinoma (n 3), hepatocellular carcinoma (n 1), liver metastases (2 colorectal, 1 renal). Five patients (62.5%) underwent surgical treatment (2 laparotomy, 2 wide nephrectomy with partial caval wall resection in HVE, 1 ex vivo liver resection with caval venoplasty in HVE and ECC). Operative mortality was 40%. Three patients underwent medical treatment (radio-chemotherapy, chemoembolization). Total survival rate was 75% at 3 months, 50% at 6 months, and 25% at 24 months. Two patients (25%) are still alive at 3 months from the diagnosis and at 36 months from the operation. PMID- 10390927 TI - [Pathological fracture of the mandible: a report of a clinical case treated with clodronate]. AB - Oral squamous cell carcinoma often invades the mandible. However, the incidence of pathological fractures of the maxillofacial bones is low and their treatment is rarely satisfactory. A patient, too weak to undergo surgery, affected by squamous cell carcinoma of the mandibular region with bone involvement and pathological fracture of the mandible, underwent chemotherapy with carboplatin associated with diphosphonate. PMID- 10390928 TI - [Laparoscopic rectopexy: our experience in the treatment of complete rectal prolapse]. AB - Surgical treatments of rectal prolapse still await a final arrangement. The aim of this work is to present Authors' experience with 12 female patients who underwent laparoscopic rectopexy. The patients, aged between 67 and 84 years, were suffering of a different degree of incontinence classified according to the Browing and Parks scale. Pneumoperitoneum was induced through the Veres needle end 5 trocars were placed. The technique used was the modified Orr-Loygue. One no death was observed and only two not serious intraoperative complications were registered, in both conversion to laparotomy was not necessary. Functional result as for incontinence has been really good (disappeared in 11 cases and improved in one). Whereas regarding the constipation, no improvement was observed in those in who in it was preexisting the operation, not appearing nevertheless, as on the contrary reported by other Authors, in those in whom it wasn't present before surgical treatment. The patients, all in follow-up (range between 10 and 36 months, average 25.08), still now experienced no relapse. In conclusion, on the base of Authors' experience, laparoscopic rectopexy is considered free of particular risks and excellent in the results even if, due to the slight number of series, any definitive judgement can be expressed. PMID- 10390929 TI - [The treatment of inguinal lymphocele and of a meta-inflammatory cyst of the abdominal wall with an injection of fibrin glue]. AB - The authors report two cases of inguinal lymphocele and phlogistic cysts of the abdominal wall treated with fibrin glue. The complete resolution of both cases after one injection, the simplicity of the technique and the low cost suggest a wider experimentation. PMID- 10390931 TI - Science and practice of indoor environment: challenges and directions? PMID- 10390930 TI - [An assessment of neuronal regeneration in neural anastomoses by synthetic guide and biological systems and insulin administration: an experimental study in the rat]. AB - The synthetic and biological nerve guide regeneration gives interesting perspective of use in making artificial conduits for peripheral nerve reconstruction. In sixty Wistar rats, under general anesthesia and with microsurgical technique, the ischiatic nerve was isolated. On the right side a segment of the nerve was removed in order to create a 10 mm gap. The defect then repaired using the conduit. Control were performed at 20, 90, 180 days and consisted in histological microscopy and electromyography investigation. The regeneration of the nerve fibers in the lumen of the conduit was not significantly different on the contralateral nerve limb. PMID- 10390932 TI - Relation between indoor and outdoor exposure to fine particles near a busy arterial road. AB - Various studies on indoor and outdoor particulate matter in the urban environment in the vicinity of busy arterial roads in the centre of the subtropical city of Brisbane have indicated that the revised United States Environmental Protection Agency National Ambient Air Quality Standards (US EPA NAAQS) for Particulate matter PM2.5 could be exceeded not only outdoors but also indoors. The aim of this work was to investigate outdoor exposure to submicrometer particles and their relationship with indoor exposure in a hypothetical office building located in the vicinity of a busy arterial road. The outdoor exposure values and trends were measured in terms of particle number in the submicrometer size range and were then recalculated to represent mass concentration trends. The results of this study indicate that exposure to PM0.7 particles in ambient air close to a busy road often exceeds the levels of the annual and 24-hour US EPA NAAQS PM2.5 standards. It is likely that exposure to PM2.5 is even higher, and may significantly exceed these standards. PMID- 10390933 TI - Quantification of birch and grass pollen allergens in indoor air. AB - Birch and grass pollen grains as well as pollen-derived small particles appear as potent allergens in the outdoor air during spring and summer. The occurrence of pollen allergens in indoor air, however, has not been studied in depth due to lack of suitable sampling and analytical methods. Herein, a recently reported "direct on sampling filter estimation" (DOSAFE) technique (Acevedo et al., 1998) has been validated for quantification of pollen allergens in indoor air using two school rooms and two office rooms as experimental models. Using DOSAFE and polyclonal antibodies against water extracts of pollen from Betula pendula and Phleum pratense L, we found that indoor air of school and office rooms carried substantial amounts of pollen allergens, expressed as SQ units, predominantly occurring as particles with smaller diameters than the pollen grains. In one school room the indoor air birch pollen allergen concentrations increased from 242 to 403 SQ units/m3 over the sampling period although the corresponding outdoor air concentrations decreased from 350 to 90 SQ units/m3. Electrostatic air cleaning in one office room reduced its grass pollen allergen concentrations by more than 95% to 0.02-0.34 SQ units/m3 as compared to the control room. PMID- 10390934 TI - Allergic and non-allergic students' perception of the same high school environment. AB - The aim of the study was to describe how allergics and non-allergics perceive the same environment. All high school students in a town in southern Sweden were invited to answer a questionnaire concerning allergy, subjective symptoms, annoyance reactions and perception of the environment (response rate: 81%). The results show that only 45% of the students were non-allergic (n = 1,715). Since the symptom frequency among non-allergic students was normal, the schools were classified as healthy. However, compared to the non-allergic students, a higher percentage among the allergics suffered from symptoms every week, a lower percentage was satisfied with the air quality and the cleaning, and a higher percentage was bothered every week by temperature, stuffy/stale air, bad odor, passive smoke, bad lighting, noise, dust and dirt (ANOVA, P < 0.05). The findings could indicate that allergics note discomfort earlier than non-allergics by being more critical in general and especially critical to factors that could effect their health. The findings could also indicate that awareness of ones own sensitivity could lead to attention to different risk factors, which in turn could lead to stress/anxiety, which could make symptoms worse. The conclusion is that it is important to take allergy into consideration when the environment is assessed. PMID- 10390935 TI - State-of-the-art in the measurement of volatile organic compounds emitted from building products: results of European interlaboratory comparison. AB - Eighteen laboratories from 10 European countries participated in a comparison organized as part of the VOCEM project, a 2.5-year research collaboration among 4 research institutes and 4 industrial companies. The scope of the project was to improve the procedure used to measure volatile organic compounds (VOC) emitted from building materials and products in small test chambers. The interlaboratory comparison included the GC-MS determination of 5 target compounds from carpet, 8 from polyvinyl chloride (PVC) cushion vinyl and 2 from paint; for the first time, chamber recovery (sinks), homogeneity of solid materials and possible contamination during transport were tested. The results show that the intralaboratory variance (random errors) is much smaller than the interlaboratory variance (systematic errors). Causes of the largest interlaboratory discrepancies were: (i) analytical errors; (ii) losses of the heaviest compounds due to sorption on the chamber walls; and (iii) non homogeneity of the materials. The output of this work concerns both the objective of labelling materials with regard to their VOC emissions and the pre-standard drafted by the European Commitee for Standardization (CEN) for this type of determination. PMID- 10390936 TI - Testing of household products and materials for emission of toluene diisocyanate. AB - Polyurethane products were subjected to chamber testing to determine their emission rates of 2,4- and 2,6-toluene diisocyanate (TDI). The polyurethane (PU) products included carpet padding, furniture cushions, sheet foam, varnishes, and sealants, as well as a commercially-applied water sealant product for concrete that contained up to 4 percent TDI by weight. The PU products were screened in a 9-L glass chamber, under elevated temperature and chamber loading conditions, using both a time-integrated sampling and analysis method specific for TDI and a continuous but non-specific real-time monitor for isocyanates. None of the products normally found in residences showed a positive response in the screening tests, indicating that TDI emissions and consequently toxic effects from such products are negligible. However, the commercially-applied water sealant gave a positive response in the screening test. Further testing of that product at realistic temperatures showed initial TDI emission rates of about 300,000 micrograms/m2/hr, with emissions lasting only one hour or less. At 21 and 27 degrees C, about 1 percent and 5 percent, respectively, of the TDI content of the product was released to the air. The emitted TDI was predominantly the 2,6 isomer, although the TDI originally present in the product was predominantly the 2,4-isomer. PMID- 10390937 TI - Room airflow studies using sonic anemometry. AB - To ensure prompt response by real-time air monitors to an accidental release of toxic aerosols in a workplace, safety professionals should understand airflow patterns. This understanding can be achieved with validated computational fluid dynamics (CFD) computer simulations, or with experimental techniques, such as measurements with smoke, neutrally buoyant markers, trace gases, or trace aerosol particles. As a supplementary technique to quantify airflows, the use of a state of-the art, three-dimensional sonic anemometer was explored. This instrument allows for the precise measurements of the air-velocity vector components in the range of a few centimeters per second, which is common in many indoor work environments. Measurements of air velocities and directions at selected locations were made for the purpose of providing data for characterizing fundamental aspects of indoor air movement in two ventilated rooms and for comparison to CFD model predictions. One room was a mockup of a plutonium workroom, and the other was an actual functioning plutonium workroom. In the mockup room, air-velocity vector components were measured at 19 locations at three heights (60, 120 and 180 cm) with average velocities varying from 1.4 cm s-1 to 9.7 cm s-1. There were complex flow patterns observed with turbulence intensities from 39% up to 108%. In the plutonium workroom, measurements were made at the breathing-zone height, recording average velocities ranging from 9.9 cm s-1 to 35.5 cm s-1 with turbulence intensities from 33% to 108%. PMID- 10390938 TI - Indoor air quality investigations at five classrooms. AB - Five classrooms, air-conditioned or naturally ventilated, at five different schools were chosen for comparison of indoor and outdoor air quality. Temperature, relative humidity (RH), carbon dioxide (CO2), sulphur dioxide (SO2), nitric oxide (NO), nitrogen dioxide (NO2), particulate matter with diameter less than 10 microns (PM10), formaldehyde (HCHO), and total bacteria counts were monitored at indoor and outdoor locations simultaneously. Respirable particulate matter was found to be the worst among parameters measured in this study. The indoor and outdoor average PM10 concentrations exceeded the Hong Kong standards, and the maximum indoor PM10 level was even at 472 micrograms/m3. Air cleaners could be used in classrooms to reduce the high PM10 concentration. Indoor CO2 concentrations often exceeded 1,000 microliters/l indicating inadequate ventilation. Lowering the occupancy and increasing breaks between classes could alleviate the high CO2 concentrations. Though the maximum indoor CO2 level reached 5,900 microliters/l during class at one of the sites, CO2 concentrations were still at levels that pose no health threats. PMID- 10390939 TI - Sick building syndrome in an office building formerly used by a pharmaceutical company: a case study. AB - In the past two decades, a group of health problems related to the indoor environment--generally termed sick building syndrome (SBS)--has emerged. We present an investigation of SBS in employees of a ministry working in a naturally ventilated office building that formerly had been used by a pharmaceutical company. A preceding environmental monitoring had failed to identify the cause(s) for the complaints. We conducted a questionnaire-based investigation and categorized the building sections and rooms according to their renovation status and their former use, respectively. The highest level of complaints was found among the employees working in rooms that in the past had been used for the production or storage of various pharmaceutical products suggesting that pharmaceutical odors may be a risk factor for SBS. Clinical laboratory tests did not show any unusual results. We conclude that the former use of a building for production and storage of pharmaceutical products should be considered as a possible risk factor for complaints about indoor air quality, e.g., when advising about or planning for renovations of buildings formerly used for production, handling, or storing of chemicals. PMID- 10390940 TI - Psychodrama groups for girls coping with trauma. AB - This study evaluated the effectiveness of psychodrama groups with traumatized middle-school girls. Comparisons of treatment and control group members' pre- and postintervention adjustment revealed significant decreases in group participants' self-reported difficulties in withdrawn behavior and anxiety/depression. Interviews with the participants reinforced the value of psychodrama group participation in the resolution of trauma and in increasing a sense of competence and self-efficacy. A brief outline of the group structure and a description of the process offer examples that illustrate the practice methodology and provide guidance for conducting psychodrama groups with vulnerable populations. Concerns with safety and containment are addressed. PMID- 10390941 TI - Reductions in criminality subsequent to group, individual, and family therapy in adolescent residential and day treatment settings. AB - The complete population of adolescents in a residential and day-treatment program over a 4-year period, 532 youths, served in two studies. Along with residential and day-treatment settings, predictive variables of interest were the number of hours spent in group, individual, and family therapy. A total of 227 adolescents qualified for Study 1 which found a reduction of rates of criminal charges from pre- to posttreatment. Study 1 also found that hours in group therapy explained the most variance in the reduction in rates of criminal charges, followed closely by hours in individual therapy. Hours in family therapy was not a significant predictor. A total of 430 adolescents qualified for Study 2, which found that residential treatment was associated with greater reductions in adult correctional commitments than day treatment. Implications stress the need for further research examining the relationships between therapeutic components of residential treatment and behavioral outcomes. PMID- 10390942 TI - Advantages of heterogeneous therapy groups in the psychotherapy of the traumatically abused: treating the problem as well as the person. AB - The authors combine a social-constructionist perspective with a psychodynamic one in discussing the problem of trauma and its treatment. They argue that effective treatment of traumatic physical, sexual, and psychological abuse must do more than alleviate the pain of the sufferer. Factors that cause and perpetuate abuse must be addressed by the abused person in conjunction with other nontraumatized persons who may have been abusive or passive in the face of abuse. An argument is made for heterogeneous therapy groups as a context for this to occur. PMID- 10390943 TI - Borderline functioning, work, and outcome in intensive evening group treatment. AB - Cluster analysis was used to identify subgroups of a sample of 40 patients with borderline personality disorder (BPD). The BPD patients were part of a larger sample that had participated in an intensive, group-oriented Evening Treatment Program. A set of pretherapy outcome measures was used to represent patient "attributes" for the cluster analysis. Eight clusters were identified. Two, each defined by a single patient with pronounced pathology, were deleted from further analyses. In a discriminant-function analysis, four dimensions emerged that differentiated the six remaining clusters. Significant relationships among the four dimensions and measures of therapeutic work and treatment outcome were identified. The relationships reflected the impact of behavioral characteristics associated with BPD on participation in and benefit from intensive group-oriented evening treatment. Implications of these exploratory findings for the understanding and treatment of BPD are discussed. PMID- 10390944 TI - Group psychotherapy treatment of borderline personalities. AB - Borderline personalities have been treated in psychotherapy groups for over 40 years. This article elaborates some of the characteristics pertinent to the treatment of these patients. Combined treatment of group and individual therapy addresses the needs for object constancy, the integration of object and self representations, and the possibility of attachment to others. Collaboration with individual therapists in this process is essential and there are specific conditions that allow this to occur as well as guidelines to help them make referrals. Cotherapy can be especially beneficial if the cotherapy team is knowledgeable and experienced. The group therapist must have special training and supervision to conduct groups of such intensity and affectively laden content. PMID- 10390945 TI - Aliveness in the work of the group: a subjective guide to creative character change. AB - The subjective response of aliveness in the work of the group may be a valuable signal on the journey toward creative character change in group therapy. In order to promote change in others, the group therapist must engage deeply and use internal responses as guides during interactions with group members as well as in relation to the group as a whole. Subjective awareness of increased aliveness that is linked with a sense of the work of the group may guide both therapist and group participants in the midst of inevitable anxieties and passions aroused during this quest for new vitality and freedom in relationships in the group. Winnicott's concept of potential space as well as group-relations theory about the primary task provide a conceptual foundation for this approach not only on the level of the individual member within the group but also on the level of the group as a whole. PMID- 10390947 TI - [Toward the 40th anniversary of "Investigacion Clinca" in the year 2000]. PMID- 10390948 TI - [Effect of climatic factors on the epidemiology of rotavirus infection in children under 5 years of age in the city of Maracaibo, Venezuela]. AB - Diarrhea is one of the most common problem of public health worldwide, specially in developing countries. In Venezuela, this affection must be weekly reported when it occurs in children under five years of age. During June 1993 to May 1995, 379 stool specimens were obtained from children under five years of age with diarrhea admitted in Hospital de Ninos of Maracaibo, Venezuela. Control group was conformed by 93 asymptomatic children. The rotavirus RNA was extracted with phenol-chloroform and precipitated with ethanol. Finally, polyacrylamide-gel electrophoresis (PAGE), followed by silver staining was employed for rotavirus detection. Our results showed 62 (16.4%) positive cases for rotavirus in symptomatic children and 9 (9.7%) in control group. The highest incidence was showed in the infants under one year of age (21.5%). The electrophoretic analysis revealed only one long electropherotype pattern in studied samples. It was observed a close relationship between the increase of rotavirus cases and the increase of the pluviometric index and a decrease of mean temperature observed during the period of study. Both variables determine the increase or decrease of viral infection in our region. This result shows the importance of the climatic factors in the rotavirus epidemiology. PMID- 10390949 TI - [Leydig cell function in experimental cryptorchism and varicocele in rats]. AB - Leydig cells were isolated from testes of normal, cryptorchid and induced- varicocele rats. These cells were counted and coincubated with and without human Chorionic Gonadotropin (hCG) during 3 hours; thereafter, steroids were measured in the incubation media. Cryptorchid animals showed the lowest number of Leydig cells, the highest Progesterone response to hCG, a slight increment of testosterone and a decrease of estradiol. On the contrary, both left and right testes from varicocele induced rats showed a higher cell number (per g of tissue), lower progesterone response, slightly higher response testosterone and lower testosterone response. These results demonstrate that these conditions of testicular hyperthermia do not affect the number and function of Leydig cells to the same degree. This may be due to differences in the testicular temperature reached with each procedure. PMID- 10390950 TI - Partial characterization of endogenous modulators of muscarinic acetylcholine receptors in human frontal cortex. AB - A soluble fraction from human frontal cortex with molecular weight less than 10 kD was tested for the presence of endogenous substances capable of modulating the [3H]-QNB binding to crude P1 + P2 fractions from the same region. The soluble fraction was able to decrease [3H]-QNB binding in a dose-response manner with an IC50 of about 30 micrograms/ml. The effect appeared to be noncompetitive in nature, since Bmax but not Kd was significantly affected; however, in some specimens a biphasic profile, with an initial inhibition of 88-90% of [3H]-QNB binding and 50-60% ulterior binding recuperation was also found. The modulator appeared to have a molecular weight less than 10,000 Daltons and was heat and trypsin resistant. These results point out the existence of an endogenous factor, which could be heterogeneous in regard to its molecular nature or to its action sites. PMID- 10390952 TI - [Achondroplasia and Down syndrome in the same patient. Report of a case]. AB - Achondroplasia and trisomy 21 are, within their respective categories, conditions the most frequent genetic diseases found in newborns. The simultaneous presence of both conditions in the same patient, has been however, reported only once in the world literature. In this paper we present a patient affected by both entities (Achondroplasia and Trisomy 21). The clinical findings and the reasons for the rare reported frequency of these cases are discussed. PMID- 10390951 TI - Women relatives of Hispanic patients with type 2 diabetes are more prone to exhibit metabolic disturbances. AB - Hyperinsulinemia and impaired insulin action are familial and predictive of Type 2 diabetes onset. Since high levels of insulin are characteristic of our general (venezuelan)hispanic population, the purpose of this investigation was to identify early metabolic defects in a group of healthy first degree relatives of Type 2 diabetic patients. We studied 46 (29 women and 17 men; ages ranging 18-66 y) first degree relatives of Type 2 diabetic patients comparing them with 22 (12 women and 10 men; ages ranging 22-60 y) subjects who had no family history of diabetes. All subjects underwent resting blood pressure and anthropometric measurements; a 75 g oral glucose tolerance test with determination of glucose and insulin and a fasting lipid profile. The relatives of Type 2 diabetic patients had higher tricipital (TC) and subscapular (SC) skinfolds, and elevated DBP in relation to the control group. The skinfolds elevation was more evident in women, while in men the elevation in DBP predominates. None of the relatives had glucose intolerance, however, the glucose-stimulated insulin response was elevated at all points in men as well as in women. No difference was observed in the HOMA values for IR and beta cell function, or in the delta I30/delta G30 ratio. The lipid profile showed a marked elevation in TG levels in men as well as in women, with low HDL-C values in men. No other lipid abnormalities were observed. Correlation analysis revealed strong association between BMI and WHR with skinfolds and several parameters of the carbohydrate metabolism in women, but not in men. IR in women was possitively associated with skinfolds, SBP and lipid parameters and beta cell function with VLDL-C. Adult relatives of Type 2 diabetic venezuelan patients from hispanic origin had, early in their lives, several parameters of the metabolic syndrome as hyperinsulinemia, obesity, dyslipidemia and high blood pressure. These alterations were more prominent in women, group in which the association among BMI, WHR and IR were statistically significant respect to SBP, DBP, basal insulin, insulin/glucose ratio, TG and HDL C. PMID- 10390953 TI - [Benzodiazepine receptor imaging in the brain: recent developments and clinical validity]. AB - Recent developments of benzodiazepine receptor imaging (123I-Iomazenil SPECT and 11C-Flumazenil PET) in neuropsychiatric disorders were reviewed. In focal epilepsy, a number of previous studies have reported a decreased benzodiazepine receptor binding in epileptic foci and greater sensitivity compared to regional cerebral blood flow imaging, especially for diagnosis of medial temporal lobe epilepsy. These findings indicate clinical validity of benzodiazepine receptor imaging in focal epilepsy and may be related to the "disinhibition mechanism" in GABA/benzodiazepine systems underlying epilepsy. In panic disorder, abnormal benzodiazepine receptor bindings are recently demonstrated in the temporal, parietal or frontal cortex. Further studies would clarify the "benzodiazepine dysfunction hypothesis" in panic disorder. PMID- 10390954 TI - [99mTc-GSA dynamic SPECT for regional hepatic functional reserve estimation: assessment of quantification]. AB - For the estimation of regional hepatic functional reserve, we produced a three dimensional hepatic functional mapping method employing 99mTc-GSA dynamic SPECT. In this analyzing protocol, we applied Patlak plot for the estimation of the liver uptake parameter of GSA, which is named hepatic GSA clearance, because it is relatively simple and suitable for matrix by matrix analysis. In this study, we assessed the physiological implication of estimated parameters and the clinical value of this analyzing method for hepatic functional reserve estimation. After venous injection of 185 MBq of GSA (3 mg), fifteen sequential sets of SPECT data were acquired for 15 minutes. First 5 sets (minutes) SPECT images were analyzed by Patlak plot and hepatic GSA clearance (ml/min) was obtained in each matrix. The sum of hepatic GSA clearance in each matrix (total hepatic GSA clearance) was calculated as an index of whole liver functional reserve. Total hepatic GSA clearance was compared with receptor index or effective blood flow (EHBF) of whole liver which were analyzed by Direct Integral Linear Least Square Regression (DILS) method for the assessment of the physiological implications of hepatic GSA clearance. The clinical value of total hepatic GSA clearance was assessed in comparisons with the conventional hepatic function test. A very good correlations were observed between total hepatic GSA clearance and receptor index (r = 0.935, p < 0.0001, n = 49), whereas the correlations between total hepatic GSA clearance and EHBF were not significant. Significant correlations were also observed between total hepatic GSA clearance and the conventional hepatic function tests, such as choline esterase (r = 0.517, p = 0.0001, n = 47), albumin (r = 0.612, p < 0.0001, n = 49), hepaplastin test (r = 0.539, p < 0.0001, n = 47), ICG R15 (r = -0.616, p < 0.0001, n = 37). These results suggested that hepatic GSA clearance strongly reflects hepatic receptor function and this parameter seemed to be useful for the hepatic functional reserve estimation. PMID- 10390955 TI - [99mTc-GSA dynamic SPECT for regional hepatic functional reserve estimation: assessment of clinical value for hepatic resection]. AB - We assessed the clinical value of three-dimensional functional mapping method employing GSA dynamic SPECT for the estimation of residual hepatic functional reserve before hepatic resection. Seventy-two consecutive patients of liver tumor were recruited in this study. Thirty-seven underwent segmentectomy or lobectomy and 35 did subsegmentectomy. GSA studies were carried out in all 72 patients before operation. Postoperative studies were performed in 70 patients about 1 month after operation, and 2 patients died of postoperative hepatic failure early after operation. In the preoperative study, liver functional images were divided into 4 segments according to liver anatomy and segmental GSA clearance was analyzed. The sum of GSA clearance of the segments immune from resection were calculated as predicted residual GSA clearance. Two patients who showed poor predicted residual hepatic GSA clearance died of postoperative hepatic failure within two months after operation (extended right lobectomy). There were good correlations between pre- and postoperative total liver clearance in patients underwent subsegmentectomy (r = 0.900, p < 0.0001, n = 35), and between predicted residual clearance and postoperative total clearance in patients underwent segmentectomy or lobectomy (r = 0.799, p < 0.0001, n = 35). After hepatic resection, there seemed to be discrepancies between hepatic volume expansion and functional restoration in some patients. Mean GSA clearance (clearance per unit volume) apparently decreased after operation in patients whose residual volume ratio (preoperative predicted residual liver volume/preoperative total liver volume) were less than 50% (p < 0.01, n = 12). These results suggested that three dimensional functional mapping method employing GSA dynamic SPECT can provide quantitative information of postoperative hepatic function and reserve before hepatic resection. Changes of mean GSA clearance after hepatic resection suggested that hepatic function per unit volume changes in the process of hepatic regeneration. PMID- 10390956 TI - [Diagnostic value of urinary N-telopeptide of type I collagen in prostate cancer: comparison with bone scintigraphy]. AB - The usefulness of a new biochemical marker of bone resorption, N-telopeptide of type I collagen (NTx), in the diagnosis of bone metastasis was assessed in 69 prostate cancer patients. Based on the bone scintigraphy findings, the patients were divided into a bone metastasis (+) group (n = 36) and a bone metastasis (-) group (n = 33). The urinary NTx level was significantly higher in the bone metastasis (+) group than in the bone metastasis (-) group (95.5 +/- 18.5 nM BCE/mM Cr vs. 63.3 +/- 7.9 nM BCE/mM Cr). There was a tendency for greater variability in urinary NTx levels during a 2 month-period in the bone metastasis (+) group than in the bone metastasis (-) group. The urinary NTx level of the 6 patients who were clinically staged as (4+) according to the extent of disease (EOD) grading system was 211.4 +/- 96.9 nM BCE/mM Cr, and was significantly higher (p < 0.05) than in the (-) group. However, there was not a significant difference in urinary NTx levels between the (1+) to (3+) groups and the (-) group. In conclusion, measuring urinary NTx levels in useful in diagnosing bone metastasis in view of the fact that it is a simple and noninvasive procedure. While it is not as sensitive as bone scintigraphy, it may be used to supplement bone scintigraphy. PMID- 10390958 TI - [Serial change of TL/BMIPP dual SPECT myocardial scintigram in patients with acute myocardial infarction; meaning of chronic mismatch phenomenon]. AB - This study was aimed to elucidate the serial changes and clinical significance of accumulation mismatch with TL and BMIPP dual SPECT myocardial scintigraphy during 6 months in patients with acute myocardial infarction (AMI). The dual SPECT scintigraphy was performed at one, three and six months after onset of AMI in 46 patients who underwent reperfusion therapy. Long axis fractional shortening in infarct-related area and left ventricular end-diastolic volume index (LVEDVI) were measured by left ventriculography performed immediately after reperfusion and at one, six months after onset of AMI. The patients were divided into two groups: those with mismatch (Group (+)) and those without (Group (-)) at one month after reperfusion. Group (+) was subdivided into three groups according to duration of persistence of mismatch; one month persistence (1 M), three months (3 M) and six months (6 M). Improvement of wall motion abnormality (WMA) in infarct related area was seen at one month after reperfusion in group 1 M and group 3 M, while group 6 M showed no apparent change in WMA throughout the study period. LVEDI did not change at six months after reperfusion in group 1 M and 3 M, while significant increase was seen in group 6 M. It is concluded that the case with disappearance of mismatch between TL and BMIPP until three months after reperfusion indicates myocardial stunning while in the case with long-standing mismatch left ventricular remodeling is suggested. PMID- 10390957 TI - [Usefulness of 18F-FDG PET for diagnosis of cardiac sarcoidosis]. AB - Cardiac sarcoidosis, the main cause of death among patients with sarcoidosis, frequently becomes clinically apparent when the disease is far advanced. To evaluate the usefulness of the 18F-fluorodeoxyglucose (FDG) positron emission tomography (PET) in detecting cardiac sarcoidosis, 18F-FDG PET was performed in 16 patients with sarcoidosis (13 female, 63 +/- 12 yrs), compared with scintigraphic findings of 99mTc-MIBI and 67Ga. Ten of 16 patients were considered to have cardiac complications on clinical grounds with tissue confirmation such as positive endomyocardial biopsy, severe ventricular arrhythmia, more than second degree atrioventricular block, and echocardiographically proven ventricular dysfunction. Among these patients with cardiac complications, abnormal myocardial uptake of FDG were observed in all (100%), which confirms significantly higher frequency compared to 67Ga scintigraphy (50%) (abnormality of 99mTc-MIBI SPECT were observed in 80%). Although abnormal FDG accumulations were observed in region with decreased uptake of 99mTc-MIBI in many cases, localization of regional abnormality of each tracer was frequently independent. This discrepancy may reflect inflammatory and degenerative process of myocardium in cardiac sarcoidosis. 18F-FDG PET is thought to be a useful noninvasive method in detecting cardiac involvement of sarcoidosis and may provide a useful information on the activity of the disease. PMID- 10390959 TI - [Assessment of ECG-gated myocardial SPECT analysis program with cardiac phantom and clinical data]. AB - PURPOSES: ECG-gated myocardial SPECT program (QGS) is coming into wide use. This program permits measurement of end-diastolic volume (EDV), and end-systolic volume (ESV) and ejection fraction (EF) by automatic detection of myocardial edges. We assessed the reproducibility, accuracy, factors that affect the measurement of these indices using a cardiac phantom and clinical data. METHODS: In the phantom study, we evaluated the effects of ventricular volume, location, absorption, acquisition time, enlarged acquisition and pre-filter on the calculated indices. In the clinical study using 99mTc-MIBI, reproducibility between 2 observers, comparison with left ventriculography and effects of pre filter were assessed. In clinical cases of 201TI and 123I-BMIPP, left ventricular volume and EF were also analyzed by QGS with various pre-filters. RESULTS: Although the true phantom volumes (y) and calculated volumes (x) showed an excellent linear correlation (y = 0.94x - 13.8, r = 0.999), calculated volumes were significantly under-estimated by 14.5-33.8%. An absorbent material around the phantom caused reduction in calculated volumes by 4.1-9.1%. Duration of acquisition times, 3 to 60 seconds per projection, did not influence the calculation of the parameters. With enlarged data collection, calculated volume (37 ml) was larger than that of normal acquisition (33 ml). When the cut-off frequency of Butterworth filter was changed, these indices of volume and EF were almost stable over 0.41 cycle/cm. There was an excellent correlation in intra observer measurements for EDV (r = 0.998, p < 0.0001), ESV (r = 0.998, p < 0.0001) and EF (r = 0.995, p < 0.0001). In comparison with left ventriculography, correlation of parameters was good in ESV (r = 0.91, p < 0.0001), EF (r = 0.88, p < 0.0001), but was fair in EDV (r = 0.78, p < 0.0001). The QGS program underestimated EDV, ESV and EF. CONCLUSION: QGS program with gated SPECT is useful to calculate relative volume and EF. However, to calculate absolute values, we should understand the various factors that affect the result of QGS. PMID- 10390960 TI - [A significance of the washout of 123I-BMIPP in patients with vasospastic angina]. AB - We investigated the washout of 123I-BMIPP from early and delayed SPECT in 28 patients with vasospastic angina from the standpoint of the intervals from the last angina attack. We divided myocardial wall into 13 segments from the early and delayed SPECT, and visually classified into four grades of defect score ranged from 0 (normal) to 3 (severe defect). Early and delayed severity scores were calculated as a total of defect scores in 3 vessel territories, and washout scores (WS) as (delayed severity score-- early severity score)/number of segments. WS of the group within 1 month from last angina attack was compared with the groups more than 1 month. In the territory of the right coronary artery, the group within 1 month showed significantly higher WS than groups more than 1 month (p < 0.05). In the other two territories, the group within 1 month showed higher WS than one of the groups more than 1 month, but the difference was not statistically significant. We considered that the washout of 123I-BMIPP may reflect the clinical course of vasospastic angina. PMID- 10390961 TI - [A method for improving the accuracy in measurement of 99mTc-GSA liver uptake]. AB - In order to improve the accuracy in the measurement of liver uptake rate of radioactivity at 15 minutes after injection of 99mTc-GSA, the corrected liver uptake rate (LU15VW) for tissue attenuation of gamma ray was measured. On the basis of the results of phantom studies, LU15VW was obtained as a ratio of the liver counts to the calculated counts of the injected dose supposed to homogeneously distribute in the liver of each patient and to decrease in count rate by the tissue attenuation of gamma ray, which was caused by the liver itself and body wall. The values of LU15VW were compared with those of the other 99mTc GSA indices (LU15, LHL15, and HH15) and of the laboratory tests of liver function in 5 patients with chronic persistent hepatitis (CPH), 25 patients with chronic active hepatitis (CAH) 2A, 8 with CAH 2B, 8 with liver cirrhosis (LC), and 20 with hepatocellular carcinoma. LU15VW showed a good correlation with LU15, LHL15, and HH15 (r = 0.912, 0.864, and -0.869). The relationships between the results of LU15VW and the laboratory tests of liver function such as ICGR15, serum albumin, platelets counts, and hepaplastin test (r = -0.800, 0.684, 0.599, and 0.465) were more excellent as compared with those between the results of the other 99mTc-GSA indices and the laboratory tests. LU15VW was distinctly different in the mean values among the three groups of patients with CAH 2A, CAH 2B, and LC. These results indicated that LU15VW was an useful method for improvement of the accuracy in the measurement of liver uptake rate of 99mTc-GSA. PMID- 10390962 TI - [Effect of mechanical damage on ex vivo DNA virus vector-mediated gene transduction in epithelial cells of murine trachea]. AB - The mechanism of Adenovirus (Ad) and Adeno-associated virus (AAV) vector-mediated gene transduction in murine tracheae has not been fully understood. Excised tracheae from mice were exposed to either Ad vector (Ad-CMV-LacZ) or AAV vector (AAV-CMV-LacZ) for 1 hour. LacZ gene expression in tracheal epithelial cells was detected by X-gal staining. Only patch distributions of LacZ expressing cells were observed. The percentage of LacZ expressing cells to total cells was less than 1% with either vector. Ad-mediated LacZ transduction was increased by mechanical damage using forceps. AAV-mediated gene transduction in tracheal epithelial cells was also increased by mechanical damage. Furthermore, this increased expression of vector LacZ by damaged epithelial cells was not affected by pretreatment with anti-ICAM-1 mAb or platelet-activating factor receptor antagonist. Although the Ad and AAV vectors were inefficient in transferring genetic material to murine trachea ex vivo, our results suggest that mechanical damage can enhance their transduction efficiency. PMID- 10390963 TI - [Changes in intrapulmonary compression gas volume during measurement of the flow volume curve]. AB - Changes in intrapulmonary compression gas volume during flow-volume curve measurement were studied in 77 patients with bronchial asthma, 20 patients with emphysema, and 14 patients with pulmonary fibrosis. The peak point of intrapulmonary compression gas volume was observed at FEF 75 in the bronchial asthma and emphysema patients, and of FEF 25 in the pulmonary fibrosis patients. In the bronchial asthma patients, intrapulmonary compression gas volume increased significantly compared to the healthy control groups; that increase was especially obvious in cases where airway obstruction was severe, e.g. when FEV 1.0% was below 69% (p < 0.05). By contrast, the emphysema patients did not exhibit any significant increase in intrapulmonary compression gas volume compared to healthy control groups despite the existence of obstructive impairments. This study demonstrated that the intrapulmonary compression gas volume-curve is a useful index for the diagnosis of pulmonary diseases when combined with measurements of FVC and FEV 1.0%. PMID- 10390964 TI - [Radiotherapy alone for elderly patients with stage III non-small cell lung cancer]. AB - We undertook a retrospective study of elderly patients with stage III non-small cell lung cancer who had been treated solely with radiotherapy during the period 1986 to 1995. Our study was designed to assess the influence of age on survival and malnutrition in patients aged 75 years or older (elderly group) and patients aged 74 years or younger (younger group). Radiotherapy alone resulted in a median survival period of 11.5 months in the younger group and 6.3 months in the elderly group (p = 0.0043). With the Cox multivariate model, good performance status, age less than 75 years, and good response were significant favorable independent predictors. Furthermore, the elderly group patients more frequently died of respiratory infections and had lower prognostic nutritional indexes than the younger group patients before and after radiotherapy. These findings suggested elderly patients with stage III non-small cell lung cancer who had been treated with radiotherapy alone had a poor prognosis and that malnutrition caused by radiotherapy was a factor contributing to the risk of death from respiratory infection in such patients. PMID- 10390965 TI - [Clinical features and outcome of pneumonia in patients with lung cancer]. AB - We reviewed our experience with pneumonia in patients with lung cancer over a 14 year period at Kurume University Hospital. We examined the clinical features and significance of pathogenic microbes isolated from sputum in patients with lung cancer complicated by pneumonia. Many investigators have noted that patients with squamous cell lung cancer tend to contract pneumonia more readily than patients with cancers of other histopathological types. Our review, however, disclosed no significant differences among histopathological types. Bacteriological examinations of sputum revealed the frequent involvement of Staphylococcus aureus, Enterococcus faecalis, and various gram-negative organisms (e.g., Pseudomonas, Acinetobacter, Enterobacter, and Klebsiella species) that are known to be causative agents of hospital-acquired infection. Beta-lactam and CLDM were less effective. Carbapenem used alone as the second regimen of treatment for lung cancer patients with pneumonia was found to be as effective as combination therapy with beta-lactam and aminoglycoside. However, more detailed investigations (e.g., randomized prospective studies) will be needed to identify suitable antibiotics against pneumonia in patients with lung cancer. We concluded that it will be necessary to evaluate the clinical features and outcome of pneumonia in lung cancer patients in order to provide more effective treatment. PMID- 10390966 TI - [Horner's syndrome in a patient with diffuse malignant pleural mesothelioma]. AB - A 63-year-old man was admitted to our hospital because of left back pain and dysesthesia in his left arm. On physical examination, the patient had ptosis, myosis, and anhydrosis on the left side, suggesting Horner's syndrome. A chest computed tomographic scan disclosed a mass lesion adjoining to the left posterior mediastinum. Although the mass lesion showed a slight decrease in size after the systemic administration of corticosteroids, no further improvement was obtained. Open chest examination revealed extended thickening of the parietal pleura with massive involvement of the upper thoracic sympathetic trunk. The diagnosis was malignant mesothelioma of sarcomatous type. Horner's syndrome is a rare but possible complication in the clinical course of malignant pleural mesothelioma. PMID- 10390967 TI - [Chronic eosinophilic pneumonia associated with rheumatoid arthritis]. AB - We encountered a 45-year-old woman with chronic eosinophilic pneumonia associated with rheumatoid arthritis. In May 1997, she was given a diagnosis of rheumatoid arthritis and prescribed non-steroidal anti-inflammatory drugs. After a month, she visited our hospital because of fever and cough. A chest roentgenogram and computed tomographic scan on first admission revealed peripheral infiltrative shadows in the upper fields of both lungs. Approximately 30% of peripheral blood cells were eosinophils. Furthermore eosinophils were elevated in bronchoalveolar lavage fluid and transbronchial lung biopsy specimens. A conclusive diagnosis of chronic eosinophilic pneumonia was made on these grounds. The patient responded well to steroid treatment, but was readmitted a week later because of worsening joint pain and skin eruptions in the lower extremities of both legs. A skin biopsy showed perivascular and interstitial eosinophil infiltration. The combination of steroids, a disease modifying anti-rheumatic drug, and a non steroidal anti-inflammatory drug proved to be effective. PMID- 10390968 TI - [A case of diffuse pleural mesothelioma disclosed by a 7 mm tumor shadow on chest X-ray film during a regular physical checkup]. AB - We encountered a patient with diffuse pleural mesothelioma pointed out by a tumor shadow 7 mm in diameter on a chest X-ray film obtained during a regular physical checkup. Computed tomography (CT) scans disclosed several small tumors, slightly thickened pleura, and a small amount of pleural effusion. A cytological examination of the effusion, as well as a bronchoscopic examination, were both unable to detect any evidence of malignancy. The patient was referred to the respiratory medicine department of our hospital, where further bronchoscopic examinations were performed but no conclusive diagnosis was obtained. The patient was therefore referred to our department. CT scans revealed enlarged tumors, markedly thickened pleura, and an increase in pleural effusion. This time a cytological analysis of effusion samples was positive for malignant mesothelioma cells. A pleuropneumonectomy was performed, but tumor cells were partially exposed on the dissected surface. Recurrent tumors were disclosed by CT scans 15 months later. Our conclusion was that a thoracoscopic examination should have been performed for diagnostic purposes at an earlier stage. PMID- 10390969 TI - [Mucosa-associated lymphoid tissue lymphoma with Sjogren's syndrome]. AB - A 68-year-old woman presented with Sjogren's syndrome. Chest X-ray films disclosed consolidated shadows in the right S2 and an infiltration shadow in the right S8 with small nodules. Pathological examination of transbronchial lung biopsy (TBLB) specimens revealed lymphocytic infiltrations that stained positive with UCHL-1 and L 26 in immunohistochemical studies. Lung tissue specimens obtained by video-assisted thoracic surgery showed lympho-epithelial lesions with dense lymphocytic infiltration. Southern blot hybridization and polymerase chain reaction (PCR) assays demonstrated monoclonality and immunoglobulin heavy chain gene rearrangement. These findings yielded a diagnosis of mucosa-associated lymphoid tissue (MALT) lymphoma. The detection of rear-ranged genes encoding for immunoglobulin heavy chains is useful for the diagnosis of primary pulmonary lymphoproliferative disorders, especially malignant lymphomas. PMID- 10390970 TI - [A case of gastric stromal tumor with chest pain and diaphragm elevation]. AB - A 66-year-old woman presented with left chest pain. Left pleural effusion was seen on a chest X-ray film and a large mass disclosed by chest computed tomography. However, the patient refused to undergo a recommended operation. Six months later, she was admitted without any symptoms. A huge (18 cm diameter) mass was detected by magnetic resonance imaging (MRI), and consisted of heterogeneous solid and cystic components. Angiography and endoscopic sonography disclosed a suspected abdominal tumor, which was resected by thoracolaparotomy. Gastric stromal tumor was diagnosed on the basis of histological findings. Chest pain and pleural effusion are rare as initial clinical symptoms of such tumors. PMID- 10390971 TI - [Interstitial pneumonia, pulmonary thrombotic microangiopathy, and hyperimmunoglobulinemia associated with Basedow's disease]. AB - A 44-year-old man with a history of Basedow's disease complained of dry cough and lymphadenopathy. Polyclonal hypergammaglobulinemia (IgG level: 5,839 mg/dl) was present, and the patient's serum was positive for lupus anticoagulant. Chest radiography disclosed irregular thickening of bronchovascular bundles, centrilobular granular and branching opacities, and small cystic changes in both lung fields. Examination of a thoracoscopic biopsy specimen revealed fibrous thickening of the pleura, interlobular septum, and peribronchiolar/perivascular connective tissue, with mild round-cell infiltration. Lymphoid follicle formation and pulmonary thrombotic microangiopathy were also observed. Corticosteroid therapy was ineffective for hypergammaglobulinemia and diffuse opacities disclosed by chest computed tomographic scans. This case differed from multicentric Castleman's disease with respect to the pathological findings of a lymph nodes biopsy, and also was not consistent with any known collagen vascular disease, (e.g., Sjogren's syndrome). We suspect this case may be representative of a new form of collagen vascular disease. PMID- 10390973 TI - [Motor neuron disease with respiratory insufficiency as primary manifestation]. AB - A 66-year-old man with dyspnea on exertion suffered a cardiac arrest and was referred to our hospital after emergency room intubation. Chest X-ray films detected no abnormalities. Blood gas analysis showed hypoxemia with normal A-aDO 2, and pulmonary function tests revealed combined ventilatory impairment. Chest fluoroscopy revealed weakness of diaphragmatic motion. No other abnormalities were found on initial examination. It was difficult to wean the patient off mechanical ventilation and identify the cause of alveolar hypoventilation. On the 60th hospital day, a neurological examination and electromyography disclosed fasciculation and denervation of the left biceps and pectoralis major muscle. These findings supported the diagnosis of motor neuron disease (MND). Although respiratory insufficiency as an initial symptom of MND is unusual, physicians should be aware of the possibility of MND in cases of alveolar hypoventilation of unknown etiology. PMID- 10390972 TI - [A case of Morgagni's foramen hernia difficult to differentiate from lipoma]. AB - A 61-year-old woman was admitted for chest discomfort. She had been admitted before, in March 1995, because of a lesion detected on chest roentgenograms. At that time, she was given a diagnosis of mediastinal lipoma based on the findings of chest computed tomography (CT) and magnetic resonance imaging (MRI), but was discharged without active intervention due to lack of subjective symptoms. During follow-up, the patient again reported chest discomfort beginning in March 1998. Because chest radiography disclosed. The tumor had enlarged, the patient was admitted to the hospital by our department. Chest MRI disclosed a mass with a signal intensity equal to that of subcutaneous fat in the pericardial space on both T1-weighted and T2-weighted images. Although sagittal images demonstrated continuity of the mass into intraperitoneal fat, a conclusive diagnosis of diaphragmatic hernia could not be made at that time. On April 30, 1998, a thoracotomy was performed on the basis of a preoperative diagnosis of mediastinal lipoma. During surgery, a hernial ring was observed slightly to the right and behind the sternum. The hernia consisted only of greater omentum, and was diagnosed as Morgagni's foramen hernia. PMID- 10390974 TI - [Successful treatment of allergic bronchopulmonary aspergillosis with erythromycin and fluconazole]. AB - A 30-year-old woman was admitted to our hospital because of productive cough, wheezing, and the disclosure of abnormal shadows on chest X-ray films. The patient was given a diagnosis of allergic bronchopulmonary aspergillosis (ABPA) based on eight findings: asthma, eosinophilia, elevated serum IgE concentrations, immediate skin reactivity to Aspergillus antigen, the presence of precipitating antibodies against Aspergillus antigen, lung infiltration, central bronchiectasis, and repeated culture of Aspergillus fumigatus in sputum. Because she refused steroids, we administered erythromycin. The volume of her sputum subsequently decreased, her symptoms were brought under control, and her serum IgE fell, but the lung infiltrates did not clear. Discontinuation of erythromycin resulted in exacerbation of the patient's asthmatic symptoms, with high fever, increased sputum volume and IgE levels, and worsening lung infiltrates. These symptoms responded well to oral prednisolone medication, but sputum culture was still positive for Aspergillus fumigatus. Following discontinuation of prednisolone, the patient was treated with erythromycin, to which oral fluconazole was added for 16 months. Subsequent sputum cultures were negative for Aspergillus fumigatus, and for 7 years thereafter the patient remained in remission. Erythromycin and anti-fungal drugs may be worth trying in cases of allergic bronchopulmonary aspergillosis. PMID- 10390975 TI - [A case of pulmonary alveolar proteinosis presenting with peripheral infiltrates]. AB - We report a case of pulmonary alveolar proteinosis (PAP). A 39-year-old asymptomatic woman was admitted to our hospital because of abnormal shadows on chest X-ray films. Chest X-ray films revealed peripheral infiltrates in both lungs. Computed tomographic examination showed patchy peripheral ground-glass attenuation, concentrated subpleurally. Bronchoalveolar lavage fluid was clear. Because transbronchial lung biopsy findings were inconclusive, a VATS-biopsy was performed. The specimens demonstrated accumulation of proteinaceous materials within alveolar spaces. The patient was given a diagnosis of PAP. Although the distribution of radiographic shadows varies in patients with PAP, perihilar or centralized shadows usually predominate. In our patient, subpleural areas of the lung were affected almost exclusively. PMID- 10390976 TI - [Molecular virology of TT virus]. AB - In 1997, a novel DNA virus was isolated from the serum of a patient with posttransfusion hepatitis of unknown etiology in Japan, and it was named TT virus (TTV) after the initials of the index patient. TTV is a nonenveloped, single stranded and circular DNA virus, and its entire nucleotide sequence of 3.9 kb has been determined. For being a DNA virus, TTV has a wide range of sequence divergence, allowing the classification into at least 16 genotypes separated by an evolutionary distance of > 0.30. Nucleotide sequence of the noncoding region is conserved, whereas the coding region sequence is highly variable. TTV strains with extremely high sequence divergence prevail in infected individuals. An association of TTV genotypes, detectable by PCR with N22 primers or genotype 1 specific primers, with hepatitis of unknown etiology suggest that TTV of restricted genotypes would be clinically important. PMID- 10390977 TI - [Molecular evolution and genotype classification of TT virus]. AB - Based on sequences data, TTV have a partial Rep protein motifs found among Circoviridae and the conserved region of parvoviral nonstructural polypeptides (NS)-1 genes, however we could not perform phylogenetic analyses among TTV and other viruses because of few similar sequences. Putative ORF2 among G2 and G4 encoded 49 aa because of in-frame stop codon, although that of G1 encoded 202 aa. Just down-stream of the stop codon, another putative new ORF(ORF3) were found around 150 aa. A phylogenetic analysis, using the ORF1 sequences of 93 TTV obtained from various geographical areas, indicated that the virus could be classified into six different genotypes. Further studies using more than 350 isolates obtained from DDBJ showed at least nine genotypes. PMID- 10390978 TI - [The association of TTV genetic variant with liver disease]. AB - TT virus (TTV), a novel DNA virus, has been reported in non-A to non-G posttransfusion hepatitis patients. Among 61 Japanese patients with liver diseases of non-B and non-C etiology, TTV DNA was detected in 15(25%) patients. The N22 region of TTV was sequenced and compared with the published sequence. Four genetic groups corresponding to G1a, G1b, G2 and G4 were formed. TTV was detected persistently regardless of its genotype/subtype. However, G2 and G4 contained 10 to 100 folds lower in titer than G1. Co-existing multiple mutants and subtypes were identified in one case. Since changes of TTV DNA titer and appearance of deduced amino acid substitution was not linked to ALT levels, the association of TTV to chronic liver diseases seemed low. PMID- 10390979 TI - [Development of the quantification method of TTV-DNA and clinical application]. AB - We developed a method to measure the quantity of TTV-DNA. This measurement is based on the principle of the real time PCR method using the TaqMan probe. By measuring the change of the fluorescent intensity caused by FRET, we could detect the amount of TTV-DNA. This method has the characteristics that the possibility of the contamination is very rare when it is compared with the usual PCR method, because the reaction system contains UNG and dUTP. This quantification method is useful for the future research of TTV to study the relationship between this virus and diseases. PMID- 10390980 TI - [Clinical characteristics and incidence in acute non A-G hepatitis]. AB - We investigated clinical characteristics and incidence of patients with acute non A-G hepatitis, who were all registered in 17 Japanese National Hospitals. Seven hundreds thirty-one (24.0%) of 3052 patients with sporadic acute hepatitis and 73 (21.2%) of 344 patients with posttransfusion acute hepatitis were diagnosed as acute non-ABC hepatitis. Patients with acute non-ABC hepatitis were older (Mean +/- SD, 44 +/- 15 years) and male/female ratio was 0.70. Although mean levels of liver function abnormality was generally mild, 4(1.8%) of 250 patients with acute non-ABC hepatitis were died of fulminant hepatitis. PMID- 10390981 TI - [Non A-G chronic hepatitis in Japan]. AB - It is clear that certain patients with viral hepatitis are also seronegative for types A, B, C, D, and E. Although hepatitis G virus was discovered recently, this virus has been reported to be non-contributing or only slightly conductive to liver dysfunction. In this article, epidemiological studies regarding patients seronegative for types A to G chronic hepatitis in Japan are reported. Among 1089 patients with chronic liver disease, twenty-five patients (1.8%; 14 chronic hepatitis, 4 cirrhosis, 2 hepatocellular carcinoma) were diagnosed as non A-G hepatitis (negative for HbsAg, HCV-Ab, and HGV RNA). Only 3 of 25 these patients had histories of blood transfusion. The levels of transaminase in the patients with chronic non-A to G hepatitis (without hepatocellular carcinoma) was as the same as those in patients with chronic hepatitis type B and C. Our results indicated a low prevalence of non-A to G chronic hepatitis, yet a few cases were progressive to cirrhosis or HCC, and this may be due to another unknown agents for non-A to G hepatitis. PMID- 10390982 TI - [Prevalence of hepatitis G virus RNA and antibodies to HGV envelope protein (anti E2) in patients with liver injury in Japan]. AB - To assess the role of hepatitis G virus (HGV) in acute and chronic liver diseases, we investigated the prevalence of HGV RNA and antibodies to HGV envelope protein (anti-E2) among patients with liver diseases diagnosed in our hospital from 1992 to 1997. Among 24 patients with acute hepatitis (HAV: 13, HBV: 2, HCV: 3, CMV: 1, non A-C: 5), only 1 patient with non A-C hepatitis (4%) were positive for HGV RNA and none was positive for anti-E2. Among 461 patients with chronic liver diseases (alcohol: 27, HBV: 74, HCV: 297, HBV + HCV: 10, non B non C: 14, autoimmune and metabolic: 39), 40 patients (alcohol: 1, HBV: 3, HCV: 33, HBV + HCV: 3) were positive for HGV RNA(9%) and 48 patients were positive for anti-E2(17%). In the patients with positive for anti-E2, only 8% were positive for HGV RNA. 98% of HGV RNA positive patients were infected with HBV or HCV, and especially 82% were infected with HCV. In patients with non A-C hepatitis, none was positive for HGV RNA, so HGV seems not to have important role in liver diseases. PMID- 10390983 TI - [Involvement of TTV, a new infectious factor in post-transfusion hepatitis, non A non G]. AB - Post-transfusion hepatitis developed in 6 of 447(1.3%) patients who received transfusion even after 1992 when HCV screening by PHA method was started. The six patients were suggested as hepatitis, non-B, non-C, non-G type. Of the six patients, 4 were found to be infected with TTV, which was identified in one of the patients. The patients became positive for TTV DNA before or almost the same time as the development of hepatitis and their amounts of TTV DNA were varied depending on the ALT level, suggesting that their hepatitis might be caused by TTV. The patients who presented hepatitis became positive for TTV DNA in the 6 10th week after transfusion, whereas more than half of the patients with a slight hepatic disorder became positive in the 2-4th week after transfusion. In addition, prolonged infection with TTV was only observed in nearly half of 23 patients. PMID- 10390984 TI - [Clinical characteristics of acute non A to G hepatitis caring TTV-DNA]. AB - We measured TTV-DNA using polymerase chain reaction method in twenty seven patients with acute non A to G hepatitis and 139 patients with known types of acute hepatitis. TTV-DNA was detected in 40.7% in the former and in 36.7% in the latter, which was not significantly different. In acute non A to G hepatitis, TTV DNA was more common in the male and in patient with more advanced age, however, no remarkable difference was noted concerning clinical characteristics according to the existence of TTV. Therefore TTV does not seem to play a causative role in pathogenesis of acute non A to G hepatitis. PMID- 10390985 TI - [Incidence of TT virus infection in patients with non-A to G liver disease]. AB - The sera of patients with liver disease associated with non-A to G hepatitis were examined for the presence of TTV DNA. These patients included 18 cases with AH, 8 cases with CH, 6 cases with LC, 4 cases with HCC, and 36 cases with blood donors. The detection of TTV DNA was performed as described by Nishizawa et al. TTV DNA was detected in 60.0%, 62.5%, 66%, 50%, 28% of the patients with AH, CH, LC and HCC, respectively. Among the patients with AH, the aminotransferases and total bilirubin values were lower in the TTV DNA-positive than -negative patients. Among the patients with chronic liver disease, however, there were no differences in the blood chemistry results between the TTV DNA-positive and -negative patients. The histological study of the liver tissues from a TTV positive patient with CH showed no evidence of necro-inflammatory reaction, although there was evidence of irregular regeneration in the TTV DNA-positive a patient. These results suggest that TTV infection may modify the pathological condition of the liver disease. PMID- 10390986 TI - [Detection of TT virus DNA in patients with non-A to G liver diseases]. AB - A novel single-stranded DNA virus, TT virus(TTV), has been reported recently. We detected TTV viral sequences by polymerase chain reaction using primers derived from nucleotide sequences of ORF1 and the 5' noncoding region of ORF2. Using primers of the 5' noncoding region, TTV DNA was detected in 21 of 25(84%) healthy individuals, suggesting that most TTV strains detected by these primers are almost harmless. In contrast, using primers of ORF1, which detect genotype 1a TTV that was reported to be a causative agent of posttransfusion hepatitis, TTV DNA was detected in only 3 of 25 healthy subjects and 3 of 27 acute and 9 of 72(12%) chronic non-A to G hepatitis patients. Whether these TTV strains actually cause hepatitis remains to be determined. PMID- 10390987 TI - [Detection of TT virus in healthy volunteer]. AB - A novel DNA virus, TT virus(TTV), has been reported in Japanese patient with non A to G posttransfusion hepatitis. We sought to determine whether TTV infection occurs in healthy volunteer, and to compared with DNA extraction methods and polymerase chain reaction(PCR) primer for TTV in diagnostic system. Using a nested PCR assay, serum sample of healthy volunteer serve our laboratory in Japan were examined for the presence of TTV DNA. Twenty of 90(22%) healthy volunteer were detected to have TTV sequences in their serum. Also, we found that DNA extraction methods with a modified phenol-chloroform method. Our result suggested that detection of TTV DNA are high ratio of adults in Japan and were necessary to take care of selected using diagnostic systems. PMID- 10390988 TI - [TTV infection in healthy subjects]. AB - Recently, a novel hepatitis related virus was described and named as TTV. The positive rates of TTV DNA in healthy subjects to far reported ranged from 1% to 62%, therefore, there must be large differences in the prevalence of TTV in healthy subjects from country to country. However, detailed data are lacking in many reports, care should be taken in analyzing them. Generally, elderly subjects tend to have increased positivity in the same area. Sanitary environment may influence the positive rate. In addition the distribution of genotype may influence the positive rate. Although the high prevalence of TTV in healthy subjects may indicate the TTV as non-hepatopathogenic virus, further studies will be needed before drawing the conclusion. PMID- 10390989 TI - [Distinct genotypes of TTV infection in non A-non C hepatitis]. AB - TTV DNA was tested by polymerase chain reaction with two set primers(NG061/NG063, T801/T935) in patients with nonA-nonC acute and chronic hepatitis. TTV DNA was detected in 75.9-100% by T801/T935, 47.1-48.3% by NG061/NG063 of 46 patients. The nucleotide sequences including 185 bp in the N22 region of 22 TTV DNA isolates from patients with acute and chronic nonA-nonC hepatitis, were classified into various TTV genotypes such as 1a, 1b and non-1. Our results demonstrate that genotype 1b may induce hepatic injury. PMID- 10390990 TI - [Positivity rate of TTV-DNA in patients with acute liver injury of undetermined etiology]. AB - To elucidate a role of TTV infection in patients with acute liver injury, TTV-DNA in the sera from 97 patients with acute liver injury of various etiology were determined according to Okamoto's method. Out of 77 patients with acute liver injury of determined etiology, 31 patients(40.3%) showed TTV-DNA positive, and out of 15 patients with acute liver injury of undetermined etiology, 8 patients(53.3%) showed TTV-DNA positive. These results suggested no evident role of TTV in patients with acute liver injury was shown. Further study including genotype and quantitative determination of TTV-DNA and antibody assay is needed. PMID- 10390991 TI - [TT virus(TTV) infection in patients with acute hepatitis]. AB - A novel DNA virus, TT virus(TTV), has been reported in patients with posttransfusion hepatitis of unknown etiology. However association between TTV and acute hepatitis has not been shown. We investigated the prevalence of TTV in acute hepatitis. TTV-positive rates in acute hepatitis A, B, C, cytomegalovirus infection, Epstein-Barr virus infection, and acute hepatitis of unknown etiology were 15.3%, 21.8%, 60.0%, 0%, 10.0%, 22.6%, respectively. There were no significant differences in TTV prevalence between each etiology and healthy blood donors(20.8%). Clinical data were similar between patients with or without TTV. In this study we could not find any difference in the prevalence of TTV between acute hepatitis with known etiologies and that with unknown etiology. TTV did not affect the clinical features of acute hepatitis with known etiologies. PMID- 10390992 TI - [TTV superinfection on acute hepatitis B]. AB - TT virus(TTV) was recently reported as candidate for a new hepatitis virus from post transfusion hepatitis of unknown etiology. In the present study, influence of TTV superinfection on acute hepatitis B was analyzed. TTV DNA was detected in sera from 10 of 44(23%) patients with acute hepatitis B, but prevalence was comparable with normal blood donor. It was unlikely that TTV superinfection affected clinical course of acute hepatitis B. In cases of TTV superinfection on hepatitis B, T. Bil and ALT values were higher than in cases of non-superinfected patients. Furthermore, HCC was appearanced in a patient of recover from acute hepatitis B. PMID- 10390993 TI - [Clinical feature of TTV-related hepatitis]. AB - It has been clarified that hepatitis G/GB virus-C is the minor cause of acute and chronic non-A-E hepatitis. But there exist non-A-E viral hepatitis patients. Recently, one of non-A-E hepatitis associated virus was identified and the new virus was named TT virus. We highly detected TT virus in the serum of hepatitis patients. TT virus were reported to be detected 1-37% of the general population in the world. TT virus may account for only a minor part of acute non-A-E hepatitis in Japan. However, whether TT virus infection really causes severe acute hepatitis is required to be elucidated. PMID- 10390994 TI - [Prevalence of TT virus in patients with fulminant hepatic failure in Japan]. AB - A novel virus(TTV) was isolated from patients with post-transfusion hepatitis of unknown etiology. We studied the prevalence of TTV in 26 patients with fulminant hepatic failure. Serum samples at admission from seven(27%) of the 26 patients were positive for TTV. TTV was also detected in 29(27%) of the 106 healthy blood donors. There were no differences in the positive rate between patients with fulminant hepatic failure and healthy blood donors. There were no differences in clinical findings, or duration from onset to coma in patients with and without TTV. However, the outcome of the patients with TTV was significant worse than those without. TTV may not cause severe hepatitis such as fulminant hepatic failure. PMID- 10390995 TI - [High prevalence of TT virus(TTV) in patients with non A to non G fulminant hepatitis: differences of clinical features and prognosis between TTV positive and negative patients]. AB - We detected TTV-DNA in sera from 36 patients with fulminant hepatitis(FH) and evaluated differences in clinical features and prognosis between TTV-DNA positive and negative patients with nonA-nonG FH. TTV-DNA in sera was measured by nested PCR. Twenty of 36 patients with FH were diagnosed nonA-nonG FH. The TTV-DNA in sera was detected in 14 patients(38.9%) with FH, 9(64%) showed nonA-nonG FH and 3 were HBV FH and 2 were drug-induced FH. Although we compared clinical features(gender, age, distribution history of blood transfusion, initial symptoms of hepatitis, and liver function tests) and prognosis between TTV positive and negative patients with nonA-nonG FH, there were no significant differences between the two groups. These data suggest that although TTV may be a infectious agent related to nonA-nonG FH, further study is needed to clarify the role of TTV in the pathogenesis of FH. PMID- 10390996 TI - [Clinical manifestations and histopathologic features of chronic liver disease with serum TT virus (TTV) DNA positive alone as measured by first generation primer sets]. AB - To determine the clinical manifestations and histopathologic features of serum TTV DNA-positive chronic liver disease, we investigated eleven patients who showed serum TTV DNA alone, as detected by the polymerase chain reaction using first generation primer sets (RD primer series). Clinical manifestations were as follows: (1) biochemical abnormalities of the ALT-dominant and gamma-GTP-dominant types; (2) persistently elevated gamma-GTP despite normalization of ALT in gamma GTP-dominant type patients; (3) association of TTV with pathogenesis of fatty liver or alcoholic liver dysfunction; and (4) response to liver-protective medicines. Histopathologic features were as follows: (1) inflammation-related cell infiltration scattered within the portal area, with distinguishable necroinflammation in the lobular region; (2) pathologic findings on biliary epithelium; (3) high incidence of steato-metamorphosis involvement; and (4) histologic characteristics undistinguishable from "viral" chronic hepatitis in some liver specimens. PMID- 10390997 TI - [Histological findings of TTV-positive non-B non-C chronic hepatitis patients]. AB - TT virus has been reported in association with patients with acute and chronic liver disease of unknown etiology. In order to estimate the pathogenesis of TTV, we investigated the liver histology in the patients of TTV-positive chronic hepatitis. The findings frequently observed in TTV-DNA positive cases were changes in liver parenchyma such as focal necrosis, hepatocyte degeneration or fatty change of various degree. On the other hand, degree of chronic inflammation in portal areas such as lymphocyte infiltration, piecemeal necrosis or fibrosis were relatively mild, which might suggest the good prognosis in TTV-positive cases. However, we could not find any differences in liver histology between TTV positive and negative patients. PMID- 10390998 TI - [The prevalence of TTV infection in non-A to G chronic liver disease, especially non-A to G hepatocellular carcinoma, and the clinical significance of TTV infection]. AB - We examined the prevalence of TT virus (TTV) infection in non-B, non-C, non-G chronic liver disease, in particular, in hepatocellular carcinoma (HCC), and the clinical significance of TTV infection. Among 829 cases with chronic liver disease, 30 cases (4%) had non-B, non-C, non-G chronic liver disease. The percentage of TTV-DNA positive cases among these 30 cases with non-B, non-C, non G chronic liver disease was 57% (17/30), significantly higher than the percentage TTV-DNA positivity (14%; 5/107) among blood donors (P < 0.01). All the TTV-DNA positive cases were still positive for TTV-DNA throughout the follow-up period (4 +/- 2 years; 1-7 years), thus demonstrating that TTV infection is persistent. These findings suggest that TTV may be one of the causes of non-B, non-C, non-G chronic liver disease. When the 30 cases of non-B, non-C, non-G chronic liver disease were divided into a TTV-DNA positive group and TTV-DNA negative group and the clinical data between the two groups compared, it was found that the serum ALP and serum gamma-GTP levels in some cases in the TTV-DNA positive group were higher than those in the TTV-DNA negative group. Twelve cases of non-B, non-C, non-G HCC were divided into two groups (TTV-DNA positive and TTV-DNA negative), and the clinical data between the two groups compared. All the four cases of HCC which were not associated with liver cirrhosis were TTV-DNA positive. However, with respect to the age at the time of onset of the HCC, no significant differences were noted between the cases of HCC associated with liver cirrhosis and those not associated with liver cirrhosis. In spite of older patients, many cases of HCC associated with TTV infection are not associated with liver cirrhosis. PMID- 10390999 TI - [A comparative study of TTV co-infection in patients with chronic liver disease B, C and non-B non-C]. AB - The TTV DNA were examined in patients with chronic liver diseases B (n = 35), C (n = 44) and non-B non-C (n = 19). The clinical background, liver function and liver histological finding were compared in patients with or without TTV infection. The prevalence of TTV in patients with chronic liver diseases B, C and non-B non-C were 37.1%, 27.3% and 52.6%, respectively. There was no significant difference in liver function test between TTV positive and negative patients with chronic liver diseases B and C. The gamma-GTP level of TTV positive patients were significantly higher than that of TTV negative patients with chronic liver diseases non-B non-C. The histological findings were similar between patients with or without TTV infection. We concluded that even though the co-infection of TTV in patients with chronic liver diseases B and C was high, TTV does not act as a liver injury agent. The relationship between TTV and non-B non-C chronic liver diseases is unclear. PMID- 10391000 TI - [Detection of TT virus (TTV) in non-B, non-C hepatitis patients with hepatocellular carcinoma, and clinical features of these patients]. AB - A novel human DNA virus, TTvirus (TTV), was identified from a patient with posttransfusion hepatitis of unknown etiology. It is thought to be a new hepatitis virus, but the clinical significance of this virus is uncertain. We investigated the frequency of TTV viremia by PCR in 39 non-B, non-C hepatitis (NBNC) patients with hepatocellular carcinoma (HCC), and clinical features of these patients. TTV viremia was detected in 20 (51.3%) of 39 NBNC hepatitis patients with HCC. Liver cirrhosis (LC) were found in 11 (55%) of 20 TTV-positive patients and 16 (84%) of 19 TTV-negative patients (p < 0.05). The levels of AST, LDH, LAP, gamma GTP in TTV-positive patients were significantly higher than those in TTV-negative patients (p < 0.05). (AST: 58 +/- 26 vs 42 +/- 23 IU/l, LDH: 468 +/- 127 vs 366 +/- 123 IU/l, LAP: 339 +/- 242 vs 206 +/- 80 IU/l, gamma GTP: 207 +/- 207 vs 105 +/- 107 IU/l) These results suggest clinical differences between TTV-positive and TTV-negative patients in NBNC hepatitis patients with HCC. PMID- 10391002 TI - [Detection of TTV DNA in hepatocellular carcinoma]. AB - A new hepatitis agent, TTV has been cloned from post-transfusion hepatitis patient sera. This virus has a single stranded DNA as genome, surrounded by viral capsid antigen without envelope. It might be a member of parvoviridae. But genome organization is far from the known parvoviridae such as B19 and adeno-associated virus. Although this virus could be a new causative agent for nonA-nonG hepatitis, high healthy carrier rate, no relation with hepatocellular carcinoma and no evidence of amplification in parencymal hepatocyte might lead to less importance in liver disease. It is necessary to accumulate further knowledge about this new agent for the coming transplantation medicine. PMID- 10391001 TI - [Prevalence of TT virus infection and viral integration in human hepatocellular carcinoma]. AB - In 1997, Nishizawa et al cloned a novel DNA virus designated as TT virus (TTV) from a patient with post-transfusion hepatitis and this virus is being thought to be a new hepatitis virus. Approximately 5 to 10% of hepatocellular carcinomas (HCCs) in Japan occurs in hepatitis B virus-negative and hepatitis C virus negative (NBNC) patients. In order to study the possible role of TTV in hepatocarcinogenesis, we studied the prevalence of the TTV DNA in liver tissue of HCC patients. As a result, TTV was shown not to be specific for NBNC HCC and TTV integration into host DNA was not detected in any HCC patient by Southern blotting. PMID- 10391003 TI - [TT virus infection in patients with hepatocellular carcinoma associated with non A to G hepatitis: histopathological study]. AB - To study if TTV infection is involved in the development of hepatocellular carcinoma (HCC), we tested the sera of 19 patients with HCC associated with non-A to G hepatitis for the presence of serum TTV DNA, and compared the blood chemistry values and liver histology of the patients in the TTV DNA-positive and negative groups. Detection of TTV DNA was performed described as Nishizawa, et al method. TTV DNA was detected in the sera of 47.4%. There were no significant differences in the blood chemistry results and other tests between the TTV positive and -negative patients. Histological examination of the non-tumor regions of the liver showed that there were no significant differences in the number of areas and characteristics of the necro-inflammatory reactions, the degree of staging and irregular regeneration of hepatocyte between the two groups. These results suggest that the development of HCC in patients with non-A to G hepatitis is not associated with TTV infection. PMID- 10391004 TI - [Virological response to interferon therapy in patients dually infected with TT virus and hepatitis C virus]. AB - In 30 chronic hepatitis C patients co-infected with TTV, TTV DNA in the sera immediately after cessation of IFN and 6 months after the end of IFN was examined. Sustained loss of TTV DNA was observed in 12 of 30 (40.0%) patients. In 7 of 30 (23.3%) patients, TTV DNA re-appeared 6 months after becoming undetectable at the end of IFN therapy. In the remaining 11 patients (36.7%), TTV DNA remained detectable during the entire follow-up period. The ALT values correlated only with the presence of HCV RNA regardless of the effect of IFN on TTV replication. This study indicates that IFN therapy is effective against TTV. PMID- 10391005 TI - [Susceptibility of TT virus to interferon therapy]. AB - A novel single stranded DNA virus, TT virus, associated with posttransfusion hepatitis was identified. The impact of the virus on development of chronic liver diseases and the susceptibility of the virus to interferon therapy has not been investigated. We retrospectively analyzed blood samples from 16 patients infected with both hepatitis C virus and TT virus who were treated by interferon. TT virus DNA was detected by nested polymerase chain reaction before, during and after interferon. Eight of these 16 patients became negative for TT virus DNA after such therapy but the virus re-appeared in one patient after cessation of therapy. All nine patients in whom interferon eliminated HCV exhibited normalization of alanine aminotransferase activity irrespective of persistence of TTV. In contrast, five of the seven patients who showed persistence of HCV after interferon had high levels of alanine aminotransferase (P = 0.034). These results suggest that TTV is sensitive to interferon and eradicated in a half of treated patients, but persists without liver injury for a long period in some patients. PMID- 10391006 TI - [TT virus infection in an area of high-endemicity for hepatitis C]. AB - TT virus (TTV) was recently identified as a candidate for a new hepatitis virus. In the present study, the clinical features and transmission routes of TTV infection were analyzed in an area highly endemic for hepatitis C virus (HCV) infection, and compared to those in an area not endemic. In conclusion, the prevalence of TTV infection was as high as 58% in the high-endemicity area for HCV infection. The main transmission route for TTV appeared to be different from that of HCV and HGV. TTV infection showed a reciprocal association with HCV infection, and had limited pathogenetic effect on hepatitis. PMID- 10391007 TI - [Seroepidemiological survey of TT virus (TTV) infection in an endemic area for hepatitis C virus]. AB - Seroepidemiological survey of TT virus (TTV) infection was performed in the inhabitants living in an endemic area for hepatitis C virus (HCV) in Gifu prefecture, Japan. In this area, 482 of 1062 inhabitants (45.4%) were seropositive for HCV relative antibodies. TTV-DNA was detected in the sera from 12 of 88 inhabitants (13.6%). The positive rate of TTV-DNA had no relation to sex or age. The TTV DNA-positive rate of the inhabitants with HCV infection was 21.4% higher than that of the inhabitants without HCV infection 10.0%. In the present survey, the rate of liver dysfunction of TTV DNA-negative inhabitants was not different from that of TTV DNA-positive inhabitants. Such results seem to suggest that TTV infection is not related to the liver disease in the area. PMID- 10391008 TI - [TTV materno-infantile infection--a study on the TTV frequency in Japanese pregnant women and the natural history of TTV mother-to-infant infection]. AB - We preliminarily describe the frequency of TTV in Japanese pregnant women, non parenteral, postnatal materno-infantile transmission of TTV, and 2 cases in which infantile development of the TTV carrier-state seemed to have occurred by vertical infection. The sera of 85 hepatitis B, C and G-positive and 36 non pathological pregnant Japanese women were screened for the presence of TTV DNA with a use of semi-nested PCR. The positive rates were 25.9 and 27.8%, respectively. No significant differences were gained between these two groups. Twenty-one infants were born to the TTV carrier women. Of them, 9 infants (42.9%) sero-converted to TTV DNA-positive after their age of 4 months. Among the infants who were breast-fed, the positive sero-conversion rate of TTV DNA tended to increase as the length of the breast feeding period increased. Serum AST/ALT levels stayed within normal upper limits in the 9 infants. This study indicates the frequent, and furthermore, certain possibility of non-parenteral (i.e., via breast milk), postnatal vertical infection of TTV. PMID- 10391009 TI - [Detection of TT virus (TTV) in Japanese hemodialysis (HD) patients]. AB - The clinical and epidemiological studies on TT virus (TTV) were achieved in 44 Japanese HD patients. TTV-DNA was detected in 29.5% of HD patients, and the percentage was higher than that reported in normal populations. HBsAg, anti-HBc, anti-HCV and HGV-RNA were positive in 6.8%, 36.4%, 22.7% and 2.3%, respectively. One of three HD patients with TTV had liver disorders. In comparison between TTV positive and negative groups, there were significant differences of anti-HBc and total cholesterol value. PMID- 10391010 TI - [Detection of TT virus in hemodialysis patients]. AB - Recently a newly discovered DNA virus, transfusion transmitted virus (TTV), was introduced as a cause of post-transfusion hepatitis. We studied the frequency of TTV viremia in 60 hemodialysis patients in Yamaguchi, Japan. TTV DNA was detected by heminested PCR, using primers described by Okamoto et al. TTV DNA was detected in 18 patients (30%). There was no differences in clinical characteristics, including age, gender, history of blood transfusion, and double infection of other hepatitis viruses, between TTV DNA positive patients and negative patients. Also the frequency of TT viremia was not associated with the duration of hemodialysis. These results suggest that the routes of TTV infection may be different from those of infection by HBV, HCV, or HGV. PMID- 10391011 TI - [The prevalence of TTV infection and the route of TTV transmission in hemodialysis patients--compared with HCV infection]. AB - Recently a novel virus named TT virus (TTV), associated with posttransfusion hepatitis, was isolated. The prevalence of TTV infection and the route of TTV transmission in HD units was investigated. TTV was detected in 51.3% of patients on HD (59/115), as compared with 16.5% of healthy blood donors (15/91). The prevalence rate of TTV in the patients without history of blood transfusion was similarly high (51.6%), compared with that of those with history of blood transfusion (51.2%). The prevalence rate of TTV did not differ according to the duration of HD. These suggest that the risk of TTV infection is very high in HD units and there is another major route of TTV transmission than blood transfusion. PMID- 10391013 TI - [Genoepidemiology and pathogenicity of TT virus in Japanese men with history of intravenous drug abuse and tattoo]. AB - Blood contamination has been proposed as TTV transmission. We studied the genoprevalence of TTV in Japanese men with history of intravenous drug abuse and/or tattoo. TTV was identified in serum by a polymerase chain reaction. TTV was detected in 89.7 percent of the men with history of intravenous drug abuse and/or tattoo, 74.4 percent of chronic hepatitis C patients, 78.0 percent of the chronic hepatitis B, and 65.8 percent of chronic hepatitis nonB nonC patients. Serum ALT levels of those infected with TTV alone were 27.2 +/- 17.5 IU/L. In the patients with chronic hepatitis C, serum ALT levels of those coinfected with TTV were similar to serum ALT levels of those without TTV infection. These results suggest that TTV causes no or mild hepatitis. PMID- 10391012 TI - [Detection rate of TTV-DNA in healthy medical workers]. AB - We examined the positive rate in healthy medical workers about TT virus (TTV) which was a new hepatitis virus reported in 1997. The healthy medical workers showed a positive rate of 24%. Because there was no significant difference of the positive rate between the medical workers and general healthy persons we could not conclude that the positive rate was influenced by their profession. Generally it is known that a positive rate changes according to PCR primers used for the measurement. As for TTV as well, it is reported that a positive rate is greatly dependent on the selected region of primer. Therefore, we must pay attention to the evaluation of the positive rate for each assay system, especially using different primers. PMID- 10391014 TI - [TT virus (TTV) in Japanese haemophiliacs]. AB - This study aimed to evaluate the prevalence and characteristics of infection with a novel virus designated TT virus (TTV) in Japanese haemophiliacs. TTV DNA was measured in 60 haemophiliacs by semi-nested PCR. HCV RNA, HGV RNA and HIV antibody were also tested. TTV DNA was detected in 35 haemophiliacs (58.3%). There were no differences in the backgrounds or characteristics between TTV DNA positive and-negative haemophiliacs, except that levels of IgG and IgM in TTV DNA positive patients were higher than those in TTV DNA-negative patients. In patients infected with TTV type 2, which is rare in Japan, the rate of coinfection with HCV of imported types was high. These indicate that TTV type 2 in Japanese haemophiliacs might have a foreign origin. PMID- 10391015 TI - [Eradication of Helicobacter pylori in Japan]. AB - Twenty five years has passed since the re-discovery of Helicobacter pylori. Many people have studied on this organism since that time. Some mechanisms about gastric mucosal inflammation have been clarified, and pathogenesis of peptic ulcer formation and gastric cancer have been solved. H. pylori infection is related to chronic gastritis, peptic ulcer, gastric carcinoma, and MALToma. In 1998, it was reported that gastric cancer occurred in H. pylori infected mongolian gerbils. In Japan, the prevalence of peptic ulcer and gastric cancer is very high. Therefore, the treatment for H. pylori infection is necessary to prevent occurrence of these diseases. To treat H. pylori infection, various regimen have been tried. Triple therapy with PPI and two antibiotics is recommended for cure of H. pylori infection in European and US guidelines. Some guidelines for management of H. pylori infection and regimen were shown in this part. PMID- 10391016 TI - Dietary carotenoids and lung cancer: a review of recent research. AB - Several hundred carotenoid research studies have been published since 1996, when two major intervention trials showed a lack of protective effect of beta-carotene supplements against lung cancer. Recent epidemiologic studies continue to show an association between high dietary intake of beta-carotene and lower risk of lung cancer. New research is attempting to clarify the apparently contradictory results of intervention and epidemiologic studies. Promising areas of investigation include characterizing biologic activities of carotenoids and gaining further insight into whether they may serve primarily as markers for a healthy lifestyle or diet. PMID- 10391017 TI - Breast-feeding, mastitis, and HIV transmission: nutritional implications. AB - In many developing countries, transmission of the human immunodeficiency virus (HIV) from mother to infant occurs through breast-feeding. Mastitis, an inflammatory process in the breast, may be common in lactating women in Africa and is associated with both higher HIV load in breast milk and mother-to-child transmission of HIV. Antioxidant micronutrient deficiencies may increase the risk of mastitis. Whether prevention, early diagnosis, and prompt treatment of mastitis will help reduce mother-to-child transmission of HIV in breast-feeding women needs further study. PMID- 10391018 TI - A genetic mutation in PPAR gamma is associated with enhanced fat cell differentiation: implications for human obesity. AB - Human obesity may have genetic causes, but determining the specific genes involved has been difficult. The peroxisome proliferator-activated receptor gamma (PPAR gamma) gene encodes a protein that plays an important role in the differentiation of fat cells. A mutation has been discovered in this gene which leads to a receptor that cannot be inactivated. This mutation, while probably rare, is associated with extreme obesity. PMID- 10391019 TI - Monitoring and evaluation of nutrition programs in developing countries. PMID- 10391020 TI - The "thrifty genotype" in 1998. PMID- 10391021 TI - Genetic variation and nutrition. PMID- 10391022 TI - Nutrition-gene interactions during intrauterine life and lactation. PMID- 10391023 TI - Genotype-environment interaction in human obesity. AB - The data reviewed in this paper reveal that individual differences in the response to alterations in energy balance induced by diet or exercise are ubiquitous. These differences are observed in a variety of obesity-related phenotypes, including body weight, body fatness, and abdominal visceral fat. Although little is known about the causes of the heterogeneity in responsiveness to dietary habits or to regular exercise, the evidence accumulated so far suggests that genetic factors may play an important role in determining the response of body mass and body fat stores to chronic alterations in energy balance. It is likely that genetic variation at several genes contributes to this heterogeneity of responses and thus to the susceptibility to obesity. Research on the genetic and molecular basis of gene-environment interactions has become a major area of investigation. One can, therefore, anticipate that major advances will occur in the coming years with respect to the identification of the genetic and molecular causes of the susceptibility to the most common diseases, including obesity. PMID- 10391024 TI - Implications of the human genome project for understanding gene-environment interactions. PMID- 10391025 TI - Lessons from genetic studies in native Canadian populations. PMID- 10391026 TI - Type 2 diabetes among the Pima Indians of Arizona: an epidemic attributable to environmental change? PMID- 10391027 TI - The Pima Indians in Sonora, Mexico. PMID- 10391028 TI - Genetics of atherosclerosis risk factors in Mexican Americans. PMID- 10391029 TI - Genetic and environmental influences on type 2 diabetes mellitus in Mexican Americans. PMID- 10391030 TI - The Mexico City Diabetes Study: a population-based approach to the study of genetic and environmental interactions in the pathogenesis of obesity and diabetes. PMID- 10391032 TI - [The use of exfoliative cytology in the detection of dysplastic changes in the laryngeal mucosa of postop cancer patients]. AB - Second primary neoplasms are the cause of failure in treatment of head and neck carcinoma in 1-34% of cases. The majority of SPM are located within mouth, pharynx, lung or oesophagus. This study was designed to evaluate to effectiveness of exfoliative cytology in postoperative follow-up of patients with carcinoma of the larynx treated with surgery. The group consisted of 81 patients who had cytological evaluation between 1994 and 1996. Cytological smears were taken from six different places in mouth, pharynx and tracheostoma. If cytomorphotic changes, parakeratosis or inflammation was found, the examination was repeated every 6 months. In the first study among 548 smears cytomorphotic changes were found in 16 (2.91%) taken from 8 patients (9.87%). 9 smears presented small or medium degree of dysplasia, 4-severe degree of dysplasia, and 3 smears demonstrated neoplastic cells. Upon completing second and third cytologic examination full remission was found in cases of small and medium dysplasia as in parakeratosis and inflammation. Stagnation was detected in cases of severe dysplasia. In the patient whose smear revealed neoplastic cells SPN of the oesophagus was diagnosed. It seems that exfoliative cytology might be a useful method for follow-up and early diagnosis of SPN in patient after laryngectomy. PMID- 10391031 TI - Obesity and cultural environment in the Yucatan region. PMID- 10391033 TI - [Surgical management of the advanced orbit malignancies]. AB - Twenty one patients with advanced malignant tumors of the orbit have been treated in the Department of Otolaryngology, Medical Academy in Warsaw, between 1980 and 1995. There were two groups of patients: five with primary orbital tumors and sixteen with secondary tumors that penetrated from sinuses. In all cases surgical treatment with orbital exenteration has been performed, as well as postoperative radiotherapy. The local cutaneus flaps and temporalis muscle flap have been used for reconstruction of the orbitomaxillary defects. The authors suggest that one stage reconstruction of maxillo-facial region after surgery decreases facial deformation and allows to apply ocular prothesis guickly. PMID- 10391034 TI - [Mechanisms smoothing deglutition disorders after oral cancer surgery]. AB - Compensatory treatment for oropharyngeal dysphagia includes postural changes and reducing the risk of aspiration. Some of compensatory maneuvers are introduced spontaneously by the ill at first weeks after oral cavity tumour resection. On the basis of roentgenotelevision examination of deglutition in 82 patients we detected mechanisms with intention to minimize swallowing disturbances. Variability of their occurrence and differences in their efficiency should be emphasized, as well as essential synchronization of laryngeal closure with emptying of pharynx and opening upper oesophageal sphincter for swallowing efficiency. The valuation of compensatory maneuvers introduced spontaneously by the ill was defined as an important part of swallowing rehabilitation. PMID- 10391035 TI - [The assessment of beta 2-microglobulin concentration in blood serum in patients with radiotherapy-treated laryngeal carcinoma]. AB - The authors present estimation of beta 2-microglobulin concentration in blood serum in patients suffering from laryngeal cancer treated with radiotherapy in the course of treatment and 3 years after. The initial concentration of beta 2 microglobulin in the blood serum was statistically significantly higher in comparison with the control group. A significantly higher initial concentration of beta 2-microglobulin was observed in the group of patients with the recurrence of cancer in relation to the group of patients without the recurrence of cancer both immediately and 3 years after radiotherapy. PMID- 10391036 TI - [Diagnostic values of nasal mucosa cytology]. AB - A semiquantitative method of the nasal smears evaluation is presented. The cytograms of healthy nasal mucosa and of various rhinitis types and nasal polyps are described. PMID- 10391037 TI - [Surgical treatment of rhinophyma using CO2 laser]. AB - The authors present the method of combined treatment of rhinophyma (surgery and laser CO2). The advantages of this method are good cosmetic result, small intraoperative bleeding and short time of healing the wound. PMID- 10391038 TI - [Surgical treatment of epistaxis]. AB - The authors present 17 patients with nose bleeding who had to be treated surgically, because conservative procedures turned out unsuccessful. Causes of bleeding, types of conservative procedures, as well as types of operations are described. Surgical treatment was fully efficacious in 12 patients (70%). PMID- 10391039 TI - [Nodular toxoplasmosis as a diagnostic and therapeutic problem]. AB - Enlargement of lymph nodes of the neck, slightly elevated body temperature and discomfort are symptoms characteristic of many illnesses. One of these can be toxoplasmosis. Because the rarity of its occurrence, sometimes toxoplasmosis may be last to be recognized. In many cases absence of specific additional examination guidelines can contribute to several problems with correct diagnosis. At the present time, the most reliable sample analysis methods are the examination of levels of antibodies IgG and IgM, and the histopathological verification. The authors also indicate that varying therapeutic effects using prophilactic treatment and insufficient additional examination could lead to diagnostic problems. PMID- 10391040 TI - [Variability of magnitude of the human jugular foramen in relation to conditions of the venous outflow after ligation of the internal jugular vein]. AB - The jugular foramina of the human skull, which are the main route of the venous outflow from the skull, are characterized by laterality. It is based upon predominance of one of the sides. This event presents considerable variability in human population. These differences are probably responsible for various results of ligation of the internal jugular vein, which is performed in some clinical situations in various patients. In order to appreciate the degree of this predominance much more precisely than it is presented in the available literature, a morphological study was carried out on 78 adult human cadavers. With an aid of digital image analysing system we measured a cross-section areas of the jugular foramina of non-macerated skull. We stated that parameters of the jugular foramina didn't differ significantly in respect to age, sex and body side. Predominance of one of the jugular foramina appeared in 83% of cases, and in remaining 17% was absent (it wasn't greater than an error of measurement). If it is present, the predominance of the left and the right side are equally probable. Laterality coefficient was also calculated. It was a ratio between cross-section area of the two jugular foramina: the greater one and the smaller one. Its amount was 1.62 in men and 1.59 in women, however this difference wasn't statistically significant. The range of the laterality was rather considerable because the coefficient differed from 1.02 to 3.6. When the degree of the laterality is high, ligation of the jugular vein on the greater side could by risky. We stated that in our material here were about 2.5% of such cases. PMID- 10391041 TI - [Hearing status after stapedectomy with preservation or cutting of stapedius tendon]. AB - The authors analysed the influence of preservation or cutting of stapedius tendon during stapedotomy on postoperative hearing improvement. The material analysed was 110 operated ears, which were divided into two egual groups considering preservation or cutting of stapedius tendon. All operations were performed by the same surgeon using stapedotomy with tarflen prothesis. In all cases before and after operation there was performed tonal audiometry within a range from 0.5 to 4.0 kHz, speech audiometry with indication of speech reception threshold using mono-syllable test. The comparison of operation results did not show significant statistical differences between the studied groups of operated ears. PMID- 10391042 TI - [Dysphagia and speech disorders]. AB - Swallowing disorders in oral and pharyngeal phase after surgery of mouth, pharynx or larynx are very often interrelated with speech and voice disorders. The results of diagnostic methods of dysphagia and voice/speech disorders based on own material of patients after total laryngectomy, partial tongue resection and cleft palate surgery were presented. Attention was also paid to other etiological factors of swallowing disorders observed in phoniatric practice. PMID- 10391043 TI - [Profound bilateral sensorineural hearing loss: a result of viral infections]. AB - A patient is presented with profound bilateral sensorineural hearing loss, as a result of viral infections. The extensive clinical diagnosis was carried out. The findings of multiple audiological and immunological tests helped to fix the proper treatment and the period of its application. In consequence, the hearing in the right ear showed a significant improvement, but the left ear wasn't recovering. The inflammatory process occurred much earlier in the left ear and caused the indivertible hearing loss. PMID- 10391044 TI - [Vestibulo-visual interaction in peripheral lesion of vestibular organ]. AB - Studies were carried out on 32 persons between 38 and 54 years of age (average 45) with unilateral peripheral lesion function of the vestibular organ (among them 8 persons with complete lack of excitability) and 10 healthy persons between 23 and 42 years of age (average 32). All of the patients were examinated for vestibulo-visual interaction. With the help of electronystagmography, the eye movement was registered after visual stimulation, vestibular stimulation and simultaneous vestibulo-visual stimulation, with impulses evoking nystagmus of direction compatible and incompatible. Movement of black stripes on the white optokinetic screen with the velocity of 15 degrees/s, to the left and to the right, was visual stimulus. Vestibular organ was activated by cold water (30 degrees C), according to the Fitzgerald Hallpike's procedure. Nystagmus reaction was evaluated in the highest intensification on the basis of the mean velocity of the slow faze, amount of nystagmus deflexion, sum of amplitudes, as well as gain. We also took into account compatibility or incompatibility of the nystagmus directions which was induced by the simultaneous of the vestibulo-visual stimulation. It has been stated that the vestibulo-visual interaction in the unilateral peripheral lesion of vestibular organ does not differ in comparison with healthy persons. Besides, simultaneous vestibulo-visual stimulation induced by the impulses incompatible in their direction causes the decrease of the parameters value of the nystagmus in healthy persons and in persons with peripheral lesion of labyrinth. In the case of one-sided fall-out function of the vestibular organ an increase was observed of value gain of optokinetic nystagmus directed at the damaged side. PMID- 10391045 TI - [Laryngological and phoniatric estimation of theological seminary alumni]. AB - Laryngological and phoniatric examinations were carried out in 130 alumni from theological seminary. The authors estimated their ability to serve the mass in the future. In 71 (54.6%) cases various pathologies of the ear, nose, oral cavity, pharynx and laryngx were found, which allowed for their early treatment and rehabilitation. It also helped to settle the limitations of their future ministration. PMID- 10391046 TI - [Assessment of otoacoustic emission usefulness for early detection of hearing impairment caused by noise]. AB - 74 miners, aged 19-35, exposed to industrial noise were examined by means of otoacoustic emission (TEOAE). The group was divided into 3 subgroups depending on the time of noise exposure. Significant weakening of otoacoustic emission in all subgroups was observed, even in subgroup with the shortest time of noise exposition and normal audiogram. The results were confirmed by others authors. Otoacoustic emission (TEOAE) could be very useful in early detection of hearing impairment cause by noise. 74 miners, aged 19-35, exposed to industrial noise were examined by means of otoacoustic emission (TEOAE). The group was divided into 3 subgroups depending on the time of noise exposure. Significant weakening of otoacoustic emission in all subgroups was observed, even in subgroup with the shortest time of noise exposition and normal audiogram. The results were confirmed by others authors. Otoacoustic emission (TEOAE) could be very useful in early detection of hearing impairment cause by noise. PMID- 10391047 TI - [Adenolymphoma (Warthin's tumor) of the larynx: coexistence with the bilateral laryngocele. Contribution to differential diagnosis with oncocytic papillary cystadenoma]. AB - Adenolymphoma is a benign monomorphic lesion composed of epithelial tissue. Usually it is found in parotid gland; however, its laryngeal occurrence was rarely described. Extraparotid Warthin's tumour as scanty lymphatic tissue. In cases when there is no lymphatic tissue in subepithelial layer it is called oncocytic cystadenoma. The authors describe the case of bifocal adenolymphoma in bilateral laryngocoele and indicate classification difficulties of oncocytic lesions in the larynx. PMID- 10391048 TI - [Laryngeal carcinoma distant metastasis to the occipital bone: a case report]. AB - The authors present a case of laryngeal carcinoma with rare metastasis to the occipital bone. PMID- 10391049 TI - [A rare case of neurofibroma in the parapharyngeal space, pharynx, skull base and neck area]. AB - Authors presented a rare case of neurofibroma involving skull base, parapharyngeal space and neck. A special emphasis was put on the preoperative diagnostic procedures and diagnostic complications as well as the complications due to the type of the operated tumor and its location. PMID- 10391050 TI - [Otolaryngological department of the Polish Military Hospital in Zeithain]. PMID- 10391051 TI - [The otolaryngological pilgrimage of Polish physicians to Vienna in the 19th century]. AB - The conditions of studying and specialistic training of Polish physicians at Vienna University in the 19th century are presented briefly. The teaching methods at Vienna University by professors, assistant professors and assistants are outlined. The origin and development of polyclinic in Vienna (Allgemeine Poliklinik) is described in detail. The therapeutical nihilism in Viennese clinics at that time is mentioned. The enumeration of such famous scientists in otiatrics and rhinolaryngology in Vienna in the 19th century as Adam Politzer, Joseph Gruber, Victor Urbantschitsch, Leopold von Schroetter, Karl Stoerk, Johann Schnitzler is pointed out finally. PMID- 10391052 TI - [Report from the International Congress of Otolaryngologists "The Danube Symposium", Bratislawa, 1998]. PMID- 10391053 TI - [Cytokines in patients with chronic renal failure during non-invasive therapy]. AB - Patients with chronic renal failure (CRF) present an immunodeficient state manifested by an abnormally high incidence of malignant tumors, enhanced susceptibility to certain infections diseases, poor responses to influenza and hepatitis B vaccines. This state coexists paradoxically with activation of most immunocompetent cells, mostly monocytes and lymphocytes. A complexed net of reciprocally acting reasons of immunological processes in patients with CRF remains still unclear. Immunological response is bounded with release and functioning of cytokines. Plasma levels of many cytokines in patients with CRF are higher than in healthy control. The main role in this process plays the state of activation of monocytes provoked by the circulating endotoxins, which has been confirmed in those patients. Moreover, chronic renal failure itself and reactive oxygen species can cause a disregulation of production and elimination of these cytokines. The decreased clearance of cytokines due to renal failure can cause an accumulation of those proteins in blood. The origin, physiological role and disordered production of cytokines needs still further investigations in order to get a better understanding of a nature of dysfunction immune system in CRF patients. PMID- 10391054 TI - [Parameters of blood acid: base balance and adequacy of continuous ambulatory peritoneal dialysis as well as dietary intake and nutritional status]. AB - The purpose of studies was: 1) an evaluation of acid-base balance parameters of capillary blood in the course of CAPD treatment with the correlation analysis between these parameters and indices of CAPD adequacy, dietary intake, nutritional status and selected clinical and laboratory findings, 2) an influence of dialysis solution containing amino acids on capillary acid-base balance. The purpose first was realized in 55 patients treated CAPD up to 24 months, the second one-in 8 CAPD patients. Kt/V, PCR, total creatinine clearance, efficacy number and clinical laboratory scores (Missouri system) were used as CAPD adequacy indices. Dietary intake was evaluated from diet histories. Indices of nutritional status included total body mass, blood concentration of total protein, albumin, Fe, ferritin and cholesterol as well as TIBC. It was shown that compensated metabolic acidosis is the most common finding in patients on CAPD. Acid-base parameters do not depend significantly on Kt/V and total creatinine clearance but there is a negative correlation between HCO3- and PCR as well as between H+ concentration and efficacy number. There is no relationship between dietary intake and acid-base parameters of CAPD patients. The worse acid-base status the greater azotemia and the higher removal of nitrogenous compounds in dialysate. It was confirmed that amino acid dialysis solution deteriorates metabolic acidosis but it can be avoided by the use of oral alkalizating drugs. PMID- 10391055 TI - [Oncologic risk of nerve sparing prostatectomy in patients with prostate cancer]. AB - At the early 80-ties Walsh introduced nerve-sparing radical prostatectomy (NSRP) which includes sparing of neurovascular bundle. This allows persistence of erection following the surgery. Benefits from this improved technique are evident, nevertheless there exist a real threat of recurrencies due to not radical removal of tumorous tissues. The aim of the present paper is to assess the oncologic risk of NSRP in own material. During 1992-1997 a total of 39 radical prostatectomies (RP) were performed, 10 of them were NSRP. RP was in each case made by retropublical approach, according to Welsh. Of the 39 patients undergoing RP a normal sexual activity before surgery was declared by 26 (66.6%). In 14 patients with preserved sexual potency the neoplastic changes were limited to single lobe. Among 10 patients treated with NSRP in 3 the changes were found in both lobes, while in one of them the infiltration tended to invade along the neurovascular bundle. In 5 patients treated with NSRP erections were preserved. On base of obtained results it is concluded that NSRP is indicated only in patients with tumors localized only in one lobe of the prostate and preferably, at the early stage of tumour development. NSRP should be performed only in those patients, who before surgery declared no troubles with erections. Oncologic risk is higher in group with NSRP. PMID- 10391056 TI - [Is polycystic kidney disease a risk factor for neoplasm formation?]. AB - Our study aimed to elicit possible association between molecular and cytogenetic damage detected in lymphocytes of three members of family with polycystic kidney disease (41 and 19 years old females and in 9 years of age boy) and predictability to develop cancer or their susceptibility to environmental exposure. The DNA damage detected by Comet assay, chromosome aberrations and sisster chromatid exchanges were tested in lymphocytes and p21ras protein level in blood plasma. Lymphocytes of two persons showed higher level of cytogenetic damages and higher in responses to 0.5 Gy dose of radiation. We think it might be associated to specific aberration present in cells of these persons. A final conclusions can be taken when an application of FISH technique would be completed. PMID- 10391057 TI - [Effect of prokinetic drugs inhibiting dopaminergic system on gallbladder motility in subjects with duodenal ulcer]. AB - The purpose of this study was to determine the effect prokinetic drugs inhibiting dopaminergic receptors on gallbladder volume in normal conditions. The study comprised 90 males, aged 20-26 years with duodenal ulcer disease in remission alloted into 3 groups, 30 subject each. The gallbladder volume was determined in group I after placebo or 10 and 20 mg or metoclopramide administration; in group II: placebo or 10 and 20 mg of domperidone and in group III after placebo or 50 and 100 mg of sulphiride administration. Gallbladder motility was assessed ultrasonographically from 15 minutes to 2 hours since placebo or drug administration. The effect of the studied prokinetic drugs in fasting on gallbladder motility was differentiated after metoclopramide no significant changes in gallbladder volume were observed; after domperidone in the dose of 20 mg a slight decrease of gallbladder volume was found, whereas after sulphiryde statistically significant decrease was detected. PMID- 10391058 TI - [Norms of arterial blood pressure for 24 hours based on biological activity]. AB - So far there is no unambiguous universally accepted standards for 24 ABPM. Aim of this study was to establish standard values of systolic and diastolic blood pressures, taking into account minimal night values, corresponding with basic biological activity. 1204 patients were tested. In 707 of them, (mean age 42.5 +/ 14.7 years), arterial hypertension was recognized. Basic on our results we determined borderline values (112/69 mmHg) which are diagnostic for arterial hypertension. Those norms eliminate influence of environmental factors on values of blood pressure. They correspond with homeostasis in hypertensive patients. PMID- 10391059 TI - [Enzymuria and urinary tract infection]. AB - Urinary tract infection (UTI) can lead to kidney damage. The analysis of urinary enzyme activities is a sensitive, non-invasive method to recognise renal tubular lesions. The most important enzymes for kidney diagnosis, pathomechanism of urinary enzyme activity changes in the course of UTI and other factors influencing enzyme excretion are described. Interpretation of enzymuria, its clinical significance for kidney damage diagnosis and prognosis of disease are presented. PMID- 10391060 TI - [Urinary tract infections in children]. AB - The advances in definition, pathogenesis, diagnosis and treatment of urinary tract infections (UTI) in children have been presented. The mechanism of UTI and predisposing factors related to host and bacteria, problems in evaluating of UTI with the use of fresh urine microscopy for the presence of leukocytes, utility and reliability of ultrasonography, intravenous urography, contrast micturating cystourethrography, and technetium-99m dimercaptosuccinic acid renoscintigraphy studies were discussed. Differences in clinical symptoms and treatment of acute and chronic phase of UTI with oral and intravenous antibiotics and other drugs depending on the age of a patient, management of asymptomatic bacteriuria, side effects of drugs, as well as prophylaxis, were reported. PMID- 10391061 TI - [Molecular structure of renin-angiotensin-aldosterone system]. AB - The application of novel investigative techniques has demonstrated that the RAA system is one of the major regulators of blood pressure and fluid and electrolyte homeostasis. There is evidence that local RAA systems exist. In this paper we have presented molecular bases of RAA system structure-genomic analysis, tissue distribution, regulation and pharmacological implications. We have payed attention to genomic linkage of the angiotensynogen gene with predisposition to the essential hypertension at human and to possibility of hormones influence at expression this gene. PMID- 10391062 TI - [Testosterone and atherosclerosis in males during andropause]. AB - Nowadays besides the commonly accepted atherogenic risk factors a special emphasis is laid on the significance of testosterone in atherogenesis in men which physiologic deficit during "andropause" is able to promote this pathology. An elevated estradiol:testosterone ratio seems to be an independent risk factor of atheromatous heart complications. There is a proved positive correlation between free testosterone, total testosterone, dihydrotestosterone and HDL cholesterol, apoA1 apolipoprotein. The relationship between LDL-cholesterol, VLDL cholesterol, total cholesterol and total and free testosterone seems to be unanimous, but in certain studies the beneficial influence of testosterone on the mentioned lipids has been observed. The discussed hormone is also functionally connected with coagulation and fibrynolisis; a positive correlation was found between endogenous testosterone and tPA-Fx and a negative correlation between testosterone and PAI-1, fibrinogen, D-dimers, alpha 2-antiplasmin. Testosterone is a functional regulator of the vascular tonus and influences on reological properties of microcirculation (the application of testosterone infusion into canine coronary arteries causes the dilatation of main and the small vessels, through NO syntetase induction and ATP-dependent K(+)- channel activation). A statistically significant positive correlation between testosterone and insulin has been stated (an elevated oestradiol:testosterone ratio is connected with the insulin resistance). Additionally a negative relationship between testosterone and android obesity has been observed. Although nowadays there are more and more facts proving the benefits of the retaining the proper testosterone levels in aging men, the final influence of the testosterone supplementary therapy on atherogenesis is not solved. PMID- 10391063 TI - [On the healthy male aging: hormonal aspects]. AB - Authors present select problems performed in the "Geneva Manifesto" published by the International Society for the Study of the Aging Male in February 1998 and pay the special attention on the inaccuracy connected with a commonly used name andropause. Additionally they present actual metabolic and clinical aspects resulting from the hormonal disturbances in aging men (hypotestosteronemia, decline in DHEA and hGH). PMID- 10391064 TI - [Safety of laser use]. AB - The paper presents the problems related to safe use of lasers being the parts of medical devices. There were discussed laser radiation interaction on biological tissues, hazards that arise during the work with laser devices, as well as fundamental safety rules and obligatory radiation protection standards. The selected constructions of anti-laser filters used for eyesight protection were also presented. PMID- 10391065 TI - [The place of radionuclide methods in diagnostic imaging of renovascular hypertension]. AB - Numerous investigations and tests are used for diagnosis of renovascular hypertension. From many imaging techniques that are applied in radiology and nuclear medicine we selected those which give us high effectiveness in diagnosing a renal artery stenosis. Choosing from the range of radiological methods we focused on ultrasonography with the "color doppler" and "power doppler" option, renal arteriography and magnetic resonance angiography. Nuclear medicine offers us renal scintigraphy and its modification--renal scintigraphy with the administration of captopril. The high sensitivity of renal scintigraphy with the use of captopril incapables us to detect a stenosis of haemodynamic significance. This is of essential value for planning surgical revascularisation procedures. PMID- 10391066 TI - [The role of dietary fiber and its preparations in the protection and treatment of overweight]. AB - Optimal amounts of dietary fibre in the diet are regarded as a protective factor against several health disorders such as some alimentary tract diseases, atherosclerosis and coronary heart disease. It is considered that the dietary fibre may help reduce body weight. The preparations of dietary fibre slow gastric emptying and decrease the appetite. However, the reduction of body weight with the application of high fibre diets, but without a change in the eating habits, is not significant. PMID- 10391067 TI - [Therapeutic vaccines in allergic diseases]. PMID- 10391068 TI - [Gastroesophageal reflux and allergic bronchial asthma. Controlled study]. AB - BACKGROUND: The association between bronchial asthma and gastroesofageal reflux provokes in the patient a reserved prognostic which if not diagnosed and treat correctly, if interferes of significative in the life quality. MATERIAL AND METHOD: With the purpose of establishing a relation between the allergic bronchial asthma of the adult, patient with digestive symptoms and healthy volunteer, is carried out a study in 162 subject with these characteristics to whom an barium esophagography was done looking for gastroesofageal reflux. RESULTS: The reflux presence in the asthmatics are of 87.23% and in the patient with digestive upsets, 39.53%, ratio odds (OR), 10-43, confidence interval (IC) of 95%, 3.38-33.83. With relation to the healthy volunteers the OR is in 11.48, IC of 95% 3.93-35.22. Among the patients with digestive upsets and the healthy volunteers the OR is of 1.10, IC of 95%, 0.68-1.64. CONCLUSIONS: The gastroesofageal reflux in the patient with bronchial asthma allergic associated to digestive sintomatology prevails more than in patient with digestive symptoms similar the asthmatics, but without bronchial asthma and also more than in the healthy volunteers. PMID- 10391069 TI - [Effect of Staphylococcus aureus extract on the expression of L-selectin and LFA 1, in neutrophils from patients with allergic bronchial asthma]. AB - OBJECTIVE: Messier L-selectin and LFA-1 in neutrophils from moderate and non atopic asthma patients, before and after stimuli with and without Sa (Staphylococcus aureus). MATERIALS AND METHOD: Design Trial; experimental. We studied neutrophils from 12 moderate and non atopic asthma patients and 12 healthy subjects before and after stimuli with and without Sa. MEASURES: The neutrophyls adhesion molecules, CD 62-L and CD 11 a was measured by citometric flow assays. RESULTS: The median of CD 62-L molecule expression increase with the stimuli in non atopic asthma patients from 2444 (CI 1966, CS 3627, RC 1661) to 6285.5 (CI 5243, CS 7203, RC 1960), and the median of CD 11 a molecule expression decrease with the stimuli in non atopic asthma patients from 9910.5 (CI 9765, CS 9961, RC 196) to 7670 (CI 7125, CS 8291, RC 1166). The median of CD 11 a molecule expression increase with the stimuli in healthy subjects from 593 (CI 361, CS 929, RC 568) to 1113 (CI 910, CS 1240, RC 330) and the median of CD 11 a molecule expression decrease with the stimuli in healthy subjects from 9850 (CI 9741, CS 9898, RC 157) to 9808.5 (CI 9693, CS 9890, RC 197) [CI. Inferior Cuartil, CS. Superior Cuartil, RC.-Cuartil Range]. PMID- 10391070 TI - [Safety and efficacy of treatment for severe atopic dermatitis with cyclosporin A and transfer factor]. AB - BACKGROUND: The atopic dermatitis is a chronic skin disease that appears in patients with a personal or family history of allergic asthma and rhinitis. It is associated to the specific activation of a gene group. In most instances, the response to the conventional treatment is adequate. The are cases, though, know as refractory, where that is not the case. The study of two therapeutic alternatives, Transfer Factor (TF) and Cyclosporin A (CyA), was elaborated for this type of patients. MATERIAL AND METHODS: Patients with severe refractory AD were studied, being admitted to the Allergic Service to the ISSSTE Lic. Adolfo Lopez Mateos, ISSSTE, between September 1997 and june 1998. They were randomly divided in two groups. The first one was subjected to CyA, on a 4 mg/kg/day dosage, with monthly surveillance of kidney and hepatic functions and blood pressure twice a week. Group two was subjected to TF, as follows: one unit every third day for the first week, two units per week for the next three weeks and one monthly unit to complete six months. Initial and final clinical and immunologic testing was performed on both groups (eosinophils, total IgE, CD4 and CD8). RESULTS: Six patients included group A, and 12 patients in group B. Both groups showed a significant statistic reduction in the total eosinophils count, without an statistic difference between them. None showed changes in the total IgE. CyA reduced the CD4 levels, while the TF increased the levels of CD8 cells, both with a p < 0.05. Both groups showed clinical improvement satistically significant, but no differences with a p > 0.05 appeared between them. Tolerance to the treatments was adequate, and there was not need to suspend the treatment in any case. Only three patients showed hypertricosis and other one presented headaches, with CyA. CONCLUSION: Both treatments showed therapeutic benefits in the treatment of patients with severe refractory AD, with similar immunologic improvement. Both drugs present different action mechanisms, so their joint application could offer clinical benefit to the patient (synergetic action), cost reduction, and long term treatments with reduced adverse effects. PMID- 10391071 TI - [Inhibitory effect of astemizol in skin tests with histamine]. AB - OBJECTIVE: To determine the effect of the astemizol to inhibit the cutaneous response to the histamine. MATERIAL AND METHODS: We made a clinical assay in healthy adult subjects realising skin tests with histamine for prick (1 mg/ml) and intradermal (0.01 mg/ml) daily during the taking of astemizol 10 mg during 7 days and during 7 days after suspending it, as well as the day 14, 21 and 28. It was determined the inhibition and the reappearance of the cutaneous response. RESULTS: They were 12 subjects with mean age of 36 years old +/- 11.2 SD. The complete inhibition was presented starting from the fourth day and most (79%) until the seventh day. The normal reaction, recovered in more than seven days but less than fourteen in 100%. CONCLUSIONS: According to these results, the astemizol inhibits the skin reaction to the histamine from the first day in 50% of the subjects and its principal action is to the seventh day, while when suspend it the normal response it recovers in more than seven days. PMID- 10391072 TI - [Effect of caffeine on tension development of skeletal muscle of mdx mouse]. AB - Among various pharmacological actions of caffeine upon the skeletal muscle, activation of the contractile system by Ca2+ is also influenced by caffeine. So far the effect of caffeine upon the contractile system is not properly studied in pathological muscle specimens. Using the skinned muscle fiber, we compared the effect of caffeine between control and dystrophic (mdx) mouse muscle. In lower Ca2+ (-5 microM), tension generation increased with caffeine. A maximum tension generation in higher Ca2+ (10-30 microM) was suppressed by 30 mM caffeine. In comparison between control and mdx type 2 muscle fiber, tension with caffeine was always more in mdx. On the other hand, modification by 5 mM caffeine was the same between control and mdx type 1 fiber of the soleus muscle. Different responses to caffeine between control and mdx type 2 fiber might be related to prevalent degenerative or regenerative processes. But exact mechanism is not clear yet. Currently, contracture by caffeine or halothane is applied in diagnosis of malignant hyperthermia (MH). Present observations suggested that influence of caffeine on the contractile system constituted one of causes for unspecific results in cases with concurrent neuromuscular diseases. PMID- 10391073 TI - [Spinal myoclonus which appeared during the administration of alpha interferon]. AB - We report a 42-year-old man who showed myoclonus in his right leg during alpha interferon (alpha-IFN) treatment for HCV-RNA positive chronic hepatitis C. The myoclonus continuously persisted even during sleeping. Surface EMG recordings revealed fairly regular about 0.3 Hz periodic grouping discharges synchronized in the right lower extremities muscles. Jerk-locked back averaging method demonstrated a monophasic negative sharp wave associated with the myoclonus. This pre-myoclonus sharp wave was localized on the Cz and preceded the myoclonus by about 60 ms. T 2-weighted gradient-echo MRI images revealed a high-intensity lesion in the right leg motor cortex in the left hemisphere. The myoclonus vanished in association with the disappearance of the above MRI lesion one month after the termination of alpha-IFN therapy. Based on these, we conclude the alpha IFN may induce the myoclonus in this patient. PMID- 10391074 TI - [Bilateral diabetic infarction of the thigh adductor muscles in a diabetic female patient-- A case report and review of the literature]. AB - A 30-year-old female complained of lancinating pain in the bilateral thighs for 10 days. The patient had a 22-year history of insulin-dependent diabetes mellitus. Physical examination revealed swelling of the bilateral lower extremities. There was exquisite tenderness on palpation over the medial thighs, with marked increase in pain on hip and knee flexion. Muscle strength of quadriceps, hamstrings, and hip adductor was decreased due to muscle pain. Pedal pulses were palpable bilaterally. Roentogenograms of the left femur revealed calcification of the left femoral arterial wall. Venogram revealed no obstruction with normal drainage. Complete blood cell count showed left shift of the neutrophils, markedly accelerated erythrocyte sedimentation rate, prolonged prothorombin time of 9 sec (normal 11.7 sec), C-reactive protein of 7.3 mg/dl and serum creatine kinase level of 175 IU/L. FBS was 225 mg/dl and Hb A 1 c was 16.4%. An MR imaging of the thighs revealed high signal intensities in the bilateral adductor muscles on T 2-weighted images. The symptoms resolved spontaneously over a three week period. From the course of the illness and MR imaging, the patient was diagnosed having diabetic muscle infarction (DMI), a rare complication of diabetes mellitus. To our knowledge, this is the first reported case of DMI in Japan. Diabetic microangiopathy and hypercoagulability are thought to be responsible for inducing DMI. Because the diagnosis can be made from the characteristic clinical and the typical MR imaging findings, muscle biopsy is not always necessary to obtain the diagnosis of DMI. PMID- 10391075 TI - [A clinico-pathological study of so-called "acute multiple sclerosis" mimicking a brain tumor on the MRI findings]. AB - We are reporting an autopsy case of so-called "acute multiple sclerosis" that was difficult to differentiate from a brain tumor on MRI findings. This case was a 69 year-old man, whose initial symptoms consisted of headache and unsteadiness in walking. Neurological findings included mild ataxia of the left upper extremity and positive Romberg sign. T 2-weighted MRI showed high intensity areas in the posterior limb of the right internal capsule and white matter near the posterior horn of the right lateral ventricle. Although the headache improved, the unsteadiness was exacerbated and the patient became unable to keep standing. Psychiatric symptoms and left hemiparesis were added to the clinical picture. The following MRI proved expansion of the previous lesions and the diffusely enhanced lesion spreading into the contralateral side through the corpus callosum. Stereotaxic biopsy showed the perivascular accumulation of small lymphocytes and a large number of bizarre astrocytes. Primary brain malignant lymphoma was diagnosed and radiation therapy was carried out. However, he developed perforation of the intestinal tract and died. Autopsy findings revealed scattered and disseminated small lesions in the cerebral white matter and the corpus callosum. There were a large number of lipid-laden macrophages, no stainable myelin and preserved axis cylinders in those lesions. Thus, those were interpreted as demyelinting lesions. They were scattered and multiple. This case was radiologically characterised by the diffusely enhanced, expanding butterfly shaped lesion in bilateral cerebral hemisphere through the corpus callosum, and pathologically proven to be acute demyelination associated with severe perivascular infiltration of inflammatory cells. Multiple sclerosis may mimic neoplastic processes as trans-callosal hyperplastic neuroimage on neuroimaging like the present case. PMID- 10391076 TI - [A case of primary central nervous system lymphoma with the onset of impotence]. AB - A 51-year-old man suffered from impotence for 10 months. Five months before he developed difficulty in urination and walking because of his both leg weakness. He was admitted to the hospital because of urinary incontinence, paraplegia and occipital headache. Neurologic examination revealed neck stiffness and Lhermitte's sign. The cranial nerves were intact with the exception of choked disc. He had weakness of lower extremities and right arm, and sensory loss on the right side including face. The plantar responses were extensor bilaterally. MR images revealed diffuse swelling in the cervical and thoracic spinal cord on a T 1-weighted image without enhancement by Gd-DTPA and diffuse high intensity of the spinal cord on a T 2-weighted image. MR image of the brain revealed the low intensity in the left temporal and occipital lobe with slightly enhancement by Gd DTPA, the high intensity in the left temporal and occipital lobe white matter on a T 2-weighted image. Diagnosis of malignant lymphoma (B cell type) was made by brain biopsy. Combined chemotherapy was performed and his symptoms resolved and the lesions on MRI disappeared. The number of cases of primary spinal intramedullary malignant lymphoma was very rare and the majority of the cases had weakness or sensory impairment in the lower extremities in the initial symptoms. This is the first case which had impotence as the initial symptom. In addition, it must be taken into consideration of this disease when the patient has myelopathy with unknown etiology. PMID- 10391077 TI - [Disappearance of essential tremor after thalamic infarction]. AB - Stereotactic thalamotomy has been used with some benefit in the treatment of essential tremor. We report a 73-year-old woman whose essential tremor of the right hand spontaneously disappeared after thalamic infarction. She had suffered hand tremor of the right hand for seven years. One morning, she noticed mild muscular weakness in her right upper and lower extremities, numbness around her mouth and paresthesia in her right arm. Simultaneously, she noticed disappearance of the tremor of her right hand. Several days later, right hemiplegia and paresthesia completely resolved. Neurological examination revealed no postural tremor or resting tremor. T 2-weighted brain MR imaging showed a high-intensity signal in the left thalamus that involved the ventralis intermedius nucleus. Clinical recovery from the effect of the infarct on essential tremor was complete. Therefore, it seems that thalamic infarction in this patient had an effect on essential tremor similar to that achieved with thalamotomy. PMID- 10391078 TI - [A case of bilateral brachial plexopathy after median sternotomy]. AB - A 53-year-old man with bilateral brachial plexopathy after median sternotomy for cardiac surgery was described. When he was awakened six days after cardiac surgery, he experienced weakness and paresthesia of both upper limbs. Neurological examination revealed moderate to severe atrophy and weakness of right biceps, brachioradialis, left triceps, and distal muscles of both upper limbs, and paresthesia around the right thumb and the left hypothenar. Deep tendon reflex of the right biceps, triceps, brachioradialis and left triceps was absent. In addition, he showed Horner's syndrome at the left side. The results of needle EMG and nerve conduction study indicated the damage of the right upper trunk and left middle and lower trunks of the brachial plexus. Although brachial plexopathy following median sternotomy has previously been reported in Western literatures, there is only a single report of unilateral brachial plexopathy in Japan. This is the first report in Japan of the bilateral brachial plexopathy following median sternotomy, and suggests that brachial plexopathy should be recognized as a complication of median sternotomy. PMID- 10391079 TI - [A case of hereditary ceruloplasmin deficiency with hemosiderosis]. AB - We report a 49-year-old female with hereditary ceruloplasmin deficiency with hemosiderosis. There was a family history of the same symptoms; her brother showed hypoceruloplasminemia and decrease of the serum copper content. On physical examinations, dementia, dysarthria, downbeat nystagmus, sensorineural hearing disturbance, orthostatic hypotension, retinitis pigmentosa, diffuse goiter, and cerebellar ataxia were noted. Laboratory examinations disclosed leukopenia, diabetes mellitus, hypothyroidism, decrease of copper content in the serum and urine. Serum ferritin concentration was remarkably increased. Serum ceruloplasmin could not be detected. Biopsy of the liver showed that iron content in the liver was increased. On MRI study, dentate nucleus of the cerebellum, basal ganglia, and the liver showed low intensity in both T1 and T2 weighted images. A nonsense mutation in the ceruloplasmin gene was found in this patient. Systemic iron deposition and tissue damage were considered as caused by deficiency of function of ceruloplasmin as ferroxidase. To our knowledge, the characteristic combination of the clinical signs in this patient has not been reported. PMID- 10391080 TI - [A case of limb-girdle type myasthenia gravis in whom rheumatoid arthritis appeared immediately after thymectomy]. AB - We report a 48-year-old female who presented limb-girdle type myasthenia gravis with inflammatory lung lesions and rheumatoid arthritis. She demonstrated a rapidly progressive muscle weakness of extremities. Neurological examination revealed facial muscle weakness, and proximal dominant limb muscle atrophy and weakness. Ptosis, ophthalmoplegia, and bulbar palsy were not observed. The edrophonium test and serum anti-acetylcholine receptor antibody were positive. The repetitive nerve stimulation showed 55% waning in the thenar muscles. From these findings, she was diagnosed as having myasthenia gravis. Plain chest X-P and body CT showed tumor-like lesions in the lung. Lung biopsy revealed the infiltration of lymphocytes. These lesions decreased in size after thymectomy and corticosteroid administration. Immediately after thymectomy, she began to have morning stiffness with pain and swelling of the finger and knee joints. RAHA test, which was negative before thymectomy, became highly positive. These findings were consistent with rheumatoid arthritis. In this patient, thymus probably played a role to suppress the development of rheumatoid arthritis. PMID- 10391081 TI - [A patient of polymyositis with severe myocardial damage and conduction block]. AB - A patient with polymyositis manifesting severe myocardial damage and conduction block is described. A 57-year-old man presented dysarthria, dysphagia, proximal dominant muscle weakness and wasting of the extremities. Muscle biopsy revealed degeneration and regeneration of muscle fibers and infiltration of mononuclear cells. After admission, muscle weakness rapidly progressed and mechanical ventilation was needed for respiratory failure. Simultaneously, cardiac symptom developed and resulted in bradycardia and trifascicular conduction block, which required a pacemaker. Echocardiogram revealed diffuse hypokinesia, ventricular enlargement and thickened wall. Marked elevations of serum CK-MB, cardiac myosin light chain I and cardiac troponin T were observed. High dose administration of methylprednisolone resulted in improvement of muscular and cardiac symptoms, and prevented complete heart block. Immediate and high dose of steroid therapy was considered to be effective for severe myocarditis in polymyositis. PMID- 10391082 TI - [Reversible posterior leukoencephalopathy in a patient receiving cyclosporin therapy]. AB - We reported a case of reversible posterior leukoencephalopathy syndrome (RPLS) that occurred during cyclosporin A (CyA) therapy for fulminant hepatitis. A 22 year-old man was given an intravenous drip of interferon-beta, metylprednisolone sodium succinate and CyA, and also received plasma exchange and hemodiafiltration. On the 7th day of the intravenous CyA therapy, in which its dose had been increased from 60 mg/day to 84 mg/day, he became somnolent and had headache, double vision, hallucination and then a generalized tonic-clonic seizure. The blood CyA concentration increased to a level as high as 455 ng/ml. Brain computed tomography (CT) scan without contrast medium revealed symmetric low-density areas in the bilateral occipital white matter and partly in the cortex. T2-weighted magnetic resonance imaging (MRI) showed an increased signal intensity, and single-photon emission CT using 99 mTc showed a hypoperfusion of cerebral blood flow in those areas. After CyA administration was changed to 100 mg/day orally to decrease its uptake in the blood, his consciousness and vision recovered within 4 weeks. Then abnormalities in MRI findings completely disappeared. On the basis of the clinical course and time-sequential change of serum CyA level in this patient, he was diagnosed as having RPLS caused by CyA therapy. Recently, the number of cases of RPLS has increased in the Western countries. However, there are few reports of RPLS after CyA therapy in Japan. From this case, we emphasize that careful following up the patient's neurological findings during CyA therapy is very important and that a cranial MRI is an essential tool for the diagnosis of RPLS. PMID- 10391083 TI - [Botulinum toxin therapy for cricopharyngeal spasm]. AB - A 36-year-old woman presented an inability to ingestion and mild hemiparesis with superficial and deep sensory disturbances on the left side for two months after a stroke. Dysphagia was originated from bilateral cricopharyngeal spasm, which was disclosed by videofluorography, manometry at the pharyngo-esophageal segment, and needle electromyography. Although no focal lesion of the brain was detected even with MRI, neurological and electrophysiological findings suggested that the lesion was localized to the lower pontine and mudullary tegmentum on the right side. Two months after the onset, botulinum toxin (10 units) was administered into the bilateral cricopharyngeal muscles, which resulted in restoration of the normal swallowing function in 5 days, and the normal function is lasting 5 years. Botulinum toxin therapy is non-invasive and effective for cricopharyngeal spasm. This non-invasive method will be the first choice for cricopharyngeal spasm replacing surgical intervention. PMID- 10391084 TI - [A case of the cerebellar atrophy and pseudo-hyperchloremia as a clue to diagnose chronic bromvalerylurea intoxication]. AB - A 36-year-old woman was admitted to our hospital because of walking disturbance and dizziness. Her neurological examination showed psychiatric disturbance, truncal and limb ataxia, wide-based gait. She has taken 3 g of bromvalerylurea daily. The laboratory test revealed elevated levels of serum chloride (201 mEq/L) and bromide (105 mg/dl), and decreased (-43 mEq/L) anion gap. Brain MRI revealed atrophy of upper part of the vermis and cerebellar hemisphere, and widening of the primary fissure and dilatation of the forth ventricle. Bromvalerylurea, which is an easily available sedative, should be still noted as a cause of cerebellar ataxia. Increased level of serum chloride was useful information for early diagnosis of chronic bromide intoxication. PMID- 10391085 TI - [A case of juvenile Parkinson disease with vocal cord abductor paralysis in the course of malignant syndrome]. AB - We report a 60-year-old woman with juvenile Parkinson disease (PD) with vocal cord abductor paralysis (VCAP). She had suffered from juvenile PD for 30 years. She was admitted in February 1998 to our clinical unit, because of malignant syndrome induced by dehydration. Neurological examination revealed disturbance of consciousness, hand tremor, dyskinesia of the trunk and all extremities, and rigidity. Laboratory examinations disclosed leukocytosis, renal dysfunction, hypermyoglobinemia, and elevation of the serum creatine kinase. Six days after admission, dyspnea and inspiratory stridor were noted, and the respiratory distress worsened. Endoscopy of the upper airways revealed that the vocal cord was in the midline or paramedian position. There are some cases of PD with VCAP, but such a case is very rare in Japan. We discussed the pathogenic mechanisms of these conditions, and speculated that VCAP was associated with malignant syndrome in our case. PMID- 10391086 TI - Silencing of the CD44 gene by CpG methylation in a human gastric carcinoma cell line. AB - We analyzed 8 human gastric carcinoma cell lines for the expression of CD44 by northern blot analysis and reverse transcription-polymerase chain reaction (RT PCR), and identified 1 cell line MKN-28 that did not express CD44. In an attempt to clarify the mechanism responsible for the inactivation of CD44 gene expression in this cell line, we investigated the methylation status around the promoter region of CD44 gene by digestion of the DNA with the methylation-sensitive restriction enzyme HpaII. The promoter region of CD44 in MKN-28 revealed hypermethylation, whereas other CD44-positive cell lines did not. Furthermore, treatment of MKN-28 with the demethylating agent 5-azacytidine restored the expression of the gene. These results suggest that CD44 expression is controlled by a DNA hypermethylation mechanism in MKN-28. PMID- 10391087 TI - Lung cancer in patients under 50 years old. AB - A long-term retrospective study was carried out on 790 cases of lung cancer to determine if the clinicopathologic characteristics and survival rates of lung cancer patients under the age of 50 differ from those of patients 50 years of age or older at diagnosis by analyzing data on patients registered at Tochigi Cancer Center Hospital. Of the 790 patients, 77 (9.7%) were under the age of 50 at diagnosis. The percentage of women in the younger patient group was significantly higher than that in the older patient group (39.0% vs. 27.5%; P = 0.034). Tumor histology revealed a significant preponderance of adenocarcinomas (60 patients, 77.9%) and a paucity of squamous cell carcinomas (8 patients, 10.4%) in the younger age group (P<0.001). The preponderance of adenocarcinoma was significant in both males and females (male: P = 0.004, female: P = 0.004). Smoking rates and rate of detection by cancer screening did not differ between the two age groups. Because of the paucity of smokers among the younger female patients, causes of lung cancer other than smoking should be sought in younger patients. No difference was found in the stage of the disease at presentation, treatment methods and survival rates between the two age groups. It is suggested that the prognosis for patients with lung cancer under the age of 50 is not significantly worse than for those aged 50 years or older, as has been shown by several investigators. PMID- 10391088 TI - Effects of D-galactosamine hydrochloride and partial hepatectomy on spontaneous hepatic injury and hepatocarcinogenesis in Long-Evans Cinnamon rats. AB - To examine the effect of nongenotoxic chemicals on hepatocarcinogenesis in Long Evans Cinnamon (LEC) rats, we gave 6-week-old male and female LEC rats (n = 18) weekly subcutaneous injections of D-galactosamine hydrochloride (GalN, 300 mg/kg) in 0.9% NaCl or only 0.9% NaCl for 50 weeks, and killed them in week 62. GalN treated male rats unexpectedly showed no lethal necrotizing hepatitis. GalN treatment increased the incidence of cholangiofibrosis in males and its severity in females, but did not cause significant increases of hepatocellular tumors in either sex. GaIN treatment increased the 5-bromo-2'-deoxyuridine (BrdU)-labeling index of hepatocytes and plasma hepatocyte growth factor, and accelerated megalocytic alterations without reduction of the hepatic copper concentration. Next, male and female LEC rats were subjected to two-thirds partial hepatectomy (PH) or sham hepatectomy in week 8 (n = 12) or in week 14 (n = 9), and killed in week 62. PH in week 14 inhibited lethal hepatitis, but PH in week 8 was less effective. PH reduced the hepatic copper concentration to half that of controls. The present data suggest that induction of hepatocyte regeneration by repeated injections of GalN, or by PH just before the onset of jaundice has a significant effect in prevention of hepatic injury of LEC rats, but not enhancement of spontaneous hepatocarcinogenesis. PMID- 10391089 TI - Increased 8-hydroxyguanine in DNA and its repair activity in hamster and rat lung after intratracheal instillation of crocidolite asbestos. AB - Asbestos and man-made-mineral fibers are known to increase one type of oxidative DNA damage, 8-hydroxyguanine (8-OH-(Gua), in vitro. In this study, we analyzed the 8-OH-Gua level in DNA and its repair activity after a single intratracheal instillation of fibers (crocidolite or glass) or saline to Syrian hamsters or Wistar rats. The 8-OH-Gua level was measured with a high-performance liquid chromatography-electrochemical detector (HPLC-ECD) system. The 8-OH-Gua repair enzyme activity was determined with an endonuclease nicking assay using a 32P labeled or fluorescently labeled 22mer DNA that contains 8-OH-Gua at a specific position. A significant increase in the 8-OH-Gua level in the lung DNA was observed 1 day after the exposure to crocidolite, as compared to the saline control. The repair activity was increased significantly at 7 days. On the other hand, after exposure to glass fibers, little or no increase of these carcinogenicity indicators was detected. These assays of 8-OH-Gua and its repair activity in short-term animal experiments will be useful for evaluating the carcinogenicity of fibers. This is the first report of the increase of 8-OH-Gua and its repair activity in the animal lung after the instillation of asbestos fibers. PMID- 10391090 TI - Mutational analysis of beta-catenin gene in Japanese ovarian carcinomas: frequent mutations in endometrioid carcinomas. AB - To investigate the contribution of the beta-catenin gene to the development of ovarian carcinomas, mutational analysis of exon 3 of the beta-catenin gene was conducted. We analyzed 61 primary ovarian carcinomas, consisting of 49 non endometrioid-type and 12 endometrioid-type tumors, for genetic alteration of the beta-catenin gene. Five carcinomas showed beta-catenin mutations (S37C, T41I, T41A), including 4 (33%) of 12 endometrioid-type tumors and 1 (14%) of 7 mucinous type tumors. All of these mutations altered at the serine/threonine residues that are potential sites of GSK3-beta phosphorylation. We detected no carcinomas with interstitial deletion involving exon 3 of beta-catenin. Furthermore, we immunohistochemically studied 27 of the 61 ovarian carcinomas. Both nuclear and cytoplasmic beta-catenin expressions were demonstrated in 4 of the 27 ovarian carcinomas for which tissue samples were available for examination. All 4 cases exhibited mutations in exon 3 of beta-catenin, including a mucinous carcinoma. Our results suggested that beta-catenin gene mutation at potential GSK3-beta phosphorylation sites results in accumulation of beta-catenin protein within the cells and its translocation to nuclei. Accumulated beta-catenin protein may be involved in the development of endometrioid-type ovarian carcinomas, and some mucinous-type ovarian carcinomas. PMID- 10391091 TI - Significance of membrane type 1 matrix metalloproteinase expression in breast cancer. AB - Expression of matrix metalloproteinases (MMPs) plays an essential role in tumor metastasis and invasion through the degradation of extracellular matrix (ECM). MT1-MMP (membrane type 1 matrix metalloproteinase), a membrane-type MMP, is responsible for the activation of MMP2. In this study the significance of MT1-MMP expression in human breast tumors was investigated by immunocytochemical assay, and its correlation with clinicobiological features was analyzed. MT1-MMP expression was detected in tumor cells and/or stromal cells, and there was a strong correlation between the expressions of MT1-MMP in the two cell types. Out of 183 primary tumors, 103 (56.2%) showed positive staining of MT1-MMP in tumor cells. MT1-MMP expression showed no significant correlation with any of the clinicobiological parameters examined, including hormone receptor status and angiogenesis. In postoperative survival analysis, MT1-MMP expression itself was not a significant prognostic factor. However, in the particular subgroup with the accumulation of thymidine phosphorylase (TP)-positive stromal cells, which have been activated by various stimuli, such as cytokines and hypoxia, MT1-MMP expression had a significant prognostic value. These data suggested that MT1-MMP might function cooperatively with tumor-associated stromal cells for the progression of breast cancer. PMID- 10391092 TI - Differentiation of a cell line of human cervical argyrophil small cell carcinoma to macrophage lineage cells. AB - To investigate the origin of argyrophil small cell carcinoma (ASCC) of the uterine cervix, we examined the influence of dibutyryl cyclic adenosine 3',5' monophosphate (dB-cAMP), a known differentiation inducer, on the characteristics of an ASCC cell line, TC-YIK, which has been shown to be a useful in vitro experimental model of ASCC. In TC-YIK cells after treatment with dB-cAMP, two specific antigenic markers of macrophages, CD14 and human leukocyte antigen-DR, were detected by flow cytometric analysis. In addition, interferon-gamma mRNA was detected by reverse transcription-polymerase chain reaction and interferon-gamma protein was detected by ELISA. More than 90% of the cells stained positive for alpha-naphthyl butyrate esterase, 1% of the cells showed phagocytotic activity against Micrococcus lysodeikticus, and 22% of the cells had M. lysodeikticus adsorbed on their surface. Furthermore, granulocyte-macrophage colony stimulating factor accelerated the proliferation of TC-YIK cells. These results indicate that dB-cAMP promotes differentiation of ASCC cells to macrophages. In contrast, less than 10% of the cells showed stellate morphology, suggesting differentiation to neuronal cells after treatment with dB-cAMP, as reported previously. Thus, TC-YIK cells have been shown to differentiate both into macrophage lineage cells and neuronal cells, suggesting that ASCC originates from undifferentiated stem cells. PMID- 10391093 TI - Immunohistochemical localization of glutathione S-transferase alpha and pi in human esophageal squamous epithelium, Barrett's epithelium and carcinoma. AB - High tissue levels of glutathione S-transferases (GSTs), a family of detoxification enzymes, are inversely correlated with cancer risk in the human gastrointestinal tract. Patients with Barrett's esophagus, wherein squamous epithelium is replaced by columnar epithelium, have an increased risk for developing esophageal adenocarcinoma. Biochemical analyses revealed that Barrett's epithelium contains lower levels of GST enzyme activity as well as some GST isoforms, as compared with squamous epithelium. So far, little information on the immunohistochemical distribution of the GST alpha and pi isoforms in normal squamous epithelium, in Barrett's metaplastic epithelium or in adeno- and squamous cell carcinomas of the esophagus is available. Tissues were fixed in formalin and embedded in paraffin. Three 4 microm thick sections were used for hematoxylin and eosin staining and for immunostaining with antibodies against GST alpha and pi. GST alpha and pi were seen in normal squamous epithelium (0% and 75%, respectively), Barrett's epithelium (75% and 100%), adenocarcinoma (25% and 100) and squamous cell carcinoma (27% and 91%). Staining was mainly cytoplasmic, though some nuclear staining with the GST pi antibody was apparent. The varying expression of GST alpha and pi in normal and (pre)neoplastic esophagus may have consequences for the treatment of these diseases and may contribute to an understanding of the development of these esophageal disorders. PMID- 10391094 TI - Reduction of end-stage malignant glioma by injection with autologous cytotoxic T lymphocytes. AB - Autologous cytotoxic T lymphocytes (CTL) against primary-cultured malignant gliomas were generated from peripheral blood mononuclear cells in vitro in 4 patients. Activities of the CTL were highly specific to the corresponding autologous glioma and were inhibited, in one patient, with antibodies against CD3, CD8 and MHC-class I molecules. When the CTL were injected 3 times into the primary-tumor-resected cavity via an Ommaya tube, reduction of the recurrent tumors with magnetic resonance imaging (MRI)-measured volumes exceeding 45 cm3 was observed in 3 patients. In a patient with glioblastoma multiforme (GBM), the tumor volume (estimated, 130 cm3) was rapidly reduced to 1/3, although re recurrence of the tumor followed 40 days later. A slight but distinct rapid reduction of the tumor volume was observed in another GBM patient and in an anaplastic astrocytoma patient; essentially no change was observed in a further GBM patient. These results suggest that adoptive immunotherapy with autologous CTL will be clinically effective against end-stage malignant gliomas. PMID- 10391095 TI - Enhancement of antiproliferative effects of interleukin-1beta and tumor necrosis factor-alpha on human prostate cancer LNCaP cells by coculture with normal fibroblasts through secreted interleukin-6. AB - The cell-cell interactions between tumor cells and stromal cells are considered to be important in the regulation of tumor development at primary and metastatic secondary sites. We studied the effects of various cytokines on the cell-cell interactions between androgen-dependent LNCaP or androgen-independent PC-3 human prostate cancer cell lines and normal fibroblasts using a co-culture system. Among the tested combinations of cytokines and fibroblasts, strong modulations of cytokine actions were seen in coculture with human normal fibroblasts WI-38. While interleukin (IL)-1beta or tumor necrosis factor-alpha (TNF-alpha) partially suppressed LNCaP cell growth in monoculture, each cytokine completely inhibited it in the case of coculture with WI-38 cells. On the other hand, they did not inhibit PC-3 cell growth significantly, regardless of monoculture or coculture. Conditioned medium prepared from WI-38 cells pretreated with IL-1beta or TNF alpha also strongly inhibited LNCaP cell growth. In the conditioned medium, marked IL-6 secretion was induced from WI-38 cells by IL-1beta or TNF-alpha. Furthermore, neutralizing antibodies to IL-6 or IL-6 receptor abrogated the antiproliferative effects of IL-1beta- and TNF-alpha-pretreated WI-38 conditioned medium. These results demonstrate that the antiproliferative effects of IL-1beta and TNF-alpha on prostate cancer cells are enhanced by coculture with normal fibroblasts through some diffusible factor(s), such as IL-6, from the stimulated fibroblasts. PMID- 10391096 TI - Manganese superoxide dismutase negatively regulates the induction of apoptosis by 5-fluorouracil, peplomycin and gamma-rays in squamous cell carcinoma cells. AB - We investigated the relationship between manganese superoxide dismutase (Mn-SOD) activity and apoptosis induced by anticancer drugs and radiation. Although the activity of copper, zinc-SOD did not differ greatly among 9 squamous cell carcinoma (SCC) cell lines (OSC-1 to OSC-9), the Mn-SOD activity did differ among the cell lines. The Mn-SOD activity was increased by treatments with 5 fluorouracil (5-FU), peplomycin and 137Cs, reaching plateau levels at 12 h after treatment and then decreasing gradually. When OSC-1 and OSC-3, and OSC-2 and OSC 4 were examined as representative cell lines with low and high Mn-SOD activity, respectively, the decrease was more prominent in OSC-1 and OSC-3 than in OSC-2 and OSC-4. The intracellular levels of superoxide and hydrogen peroxide (H2O2) were increased after treatment with the anticancer agents, and the increases were larger in OSC-1 and OSC-3 than in OSC-2 and OSC-4. The decrease of mitochondrial membrane potential (deltapsi(m)) by the anticancer agents was marked in OSC-1 and OSC-3. Correspondingly, the release of cytochrome c, the activation of caspase-3 and the cleavage of poly(ADP-ribose)polymerase were stronger in OSC-3 than in OSC 4. In addition, apoptosis induced by the anticancer agents was prominent in OSC 3, exhibiting a close relationship with the deltapsi(m) and the H2O2 level. These results indicate that Mn-SOD in SCC cells modulates apoptosis induction and the inactivation of Mn-SOD might be a promising strategy for SCC treatment. PMID- 10391097 TI - N-acetylcysteine modifies cis-dichlorodiammineplatinum-induced effects in bladder cancer cells. AB - We previously demonstrated a role of reactive oxygen species (ROS) in cytotoxicity induced by cis-dichlorodiammineplatinum (CDDP) in combination with glutathione (GSH) depletors in bladder cancer cells. However, the relationship between CDDP and ROS is still unclear, although many mechanisms of drug resistance have been well characterized. The present study was undertaken to investigate the effects of N-acetylcysteine (NAC), a GSH precursor, on the CDDP induced effects in bladder cancer cells (KU1). The cytotoxic effects of CDDP were significantly blunted by NAC (1 mM) in KU1 cells. The IC50 of CDDP only (10.2+/ 1.2 microM) is significantly lower than that of CDDP with NAC (IC50: 20.3+/-1.6 microM) in KU1 cells. NAC also significantly increased the intracellular concentration of GSH in KU1 cells (37.2+/-1.6 nmol/10(6) cells), compared to controls (15.9+/-7.6 nmol/10(6) cells). While CDDP produced a significant increase in ROS as measured in terms of dichlorofluorescein (DCF) production in KU1 cells in a time-dependent manner, pretreatment with NAC significantly reduced CDDP-induced intracellular DCF in KU1 cells. Moreover, TdT-mediated dUTP-biotin nick-end labeling (TUNEL) assay showed that CDDP-induced apoptosis (31.1+/-3.8%) was significantly inhibited by pretreatment with NAC in KU1 cells (11.2+/-2.6%). These results demonstrated that NAC scavenges CDDP-induced ROS and inhibits CDDP induced cytotoxicity, suggesting that ROS mediate the CDDP-induced cytotoxicity in bladder cancer cells. PMID- 10391098 TI - Regulatory network of mitomycin C action in human colon cancer cells. AB - A network composed of activation and inactivation pathways to regulate mitomycin C (MMC) action is suggested to exist in human cancer cells. COLO201 colon cancer cells were stably transfected with human NQO1 cDNA that encodes NAD(P)H:quinone oxidoreductase (DT-diaphorase, DTD), and a clonal cell line with about 57-fold elevated DTD activity was obtained. Northern analysis revealed that expression of the NADPH:cytochrome P450 reductase (P450 reductase) gene was decreased in the transfectant, COLO201/NQO1, associated with the increase of NQO1 expression. Biochemical characterization of the cells showed a significant increase of the glutathione (GSH) content concomitantly with the decrease of the P450 reductase activity. As a result of these coordinated modulations, sensitivity of COLO201/NQO1 to MMC was not increased as compared to the parent cells. Analyses of inhibition by specific inhibitors of DTD, P450 reductase and glutathione S transferase (GST) in 5 human colon cancer cell lines including the transfectant showed that DTD and P450 reductase play significant roles in MMC activation in cells with sufficiently high DTD activity and with marginal DTD activity, respectively. In contrast, GST appeared to participate in MMC inactivation in cells with a high level of GST activity. These results indicated that DTD, P450 reductase, GSH and GST may act together compensatively or competitively, depending on their levels in cells, to determine the cellular sensitivity to MMC. PMID- 10391100 TI - High-dose therapy and stem cell transplantation in follicular lymphoma. AB - Indolent follicular lymphomas are diseases which are generally incurable with conventional therapy. Although patients can survive for prolonged periods, the median duration of first remissions is about 2.5 years, and subsequent remissions progressively shorten with time. High-dose therapy with hematopoietic stem cell support leads to prolonged disease-free and overall survival in a subset of patients with aggressive non-Hodgkin's lymphoma. Mounting evidence suggests similar findings for selected patients with indolent follicular non-Hodgkin's lymphoma. It is still unclear as to when this approach should be used; however, inferior results have been seen in heavily pretreated patients. In contrast, encouraging results are being reported in patients undergoing such treatment early in the course of their disease. Despite these data, many patients continue to relapse, and investigations are now focused on eradication of minimal residual disease, allogeneic transplantation, novel ablative regimens, and improvements in stem cell purging. PMID- 10391099 TI - In vitro antitumor activity of TAS-103, a novel quinoline derivative that targets topoisomerases I and II. AB - TAS-103 is a novel anticancer agent targeting both topoisomerase (Topo) I and Topo II, that stabilizes cleavable complexes of Topo-DNA at the cellular level. In this study, the in vitro antitumor effects of TAS-103 were compared with those of other known Topo I and Topo II inhibitors. TAS-103 inhibited DNA synthesis more strongly than RNA and protein synthesis, and induced an increase of cell population in the S-G2/M phase. The cytotoxicity of TAS-103 was strongest against S-phase cells, but its cell cycle phase specificity was not clear, and depended on drug concentration and exposure time. The cytotoxicity of TAS-103 (IC50: 0.0030-0.23 microM) against various tumor cell lines was much stronger than that of VP-16 and comparable to that of SN-38. The cytotoxicity of TAS-103 seemed to be more related to the amount of protein-DNA complexes than to the accumulation of TAS-103 in the cells. P-Glycoprotein (P-gp)-mediated MDR, CDDP-resistant and 5 FU-resistant cell lines did not show cross-resistance to TAS-103. Although PC 7/CPT cells bearing a Topo I gene mutation showed cross-resistance to TAS-103, the sensitivity of P388/CPT, HT-29/CPT and St-4/CPT cells, showing decreased Topo I expression, was not changed. KB/VM4 and HT-29/Etp cells, showing decreased Topo II expression, were slightly cross-resistant to TAS-103. These results suggest that TAS-103 may act as an inhibitor of both Topo I and Topo II at the cellular level. This property may be responsible for its strong antitumor effect and broad spectrum, growth-inhibitory effect on drug-resistant cell lines. PMID- 10391101 TI - An efficient retrovirus-mediated transduction of human blood coagulation factor VIII cDNA in regenerating rat liver. AB - A retrovirus-mediated transduction of B-domain-deleted human blood coagulation factor VIII (FVIII-B) was attempted in partially hepatectomized rats. FVIII-B cDNA was inserted into a retroviral vector (pLNSX) and infective recombinant virus particles were produced in packaging cell lines (psi2 and PA317). Transfection of mouse NIH-3T3 cells with the FVIII-B cDNA inserted recombinant viruses, followed by G418 selection, gave a viral titer of 3.5 x 10(4) CFU/ml. FVIII-B protein, as well as FVIII-B mRNA, was detected in these cells. Transfusion of FVIII-B-expressing retrovirus particles into the tail vein of rats subjected to partial hepatectomy resulted in a relatively higher level of FVIII-B expression in liver and circulating plasma as compared with the sham-operated rats. These results indicate that the augmentation of FVIII activity in the blood of an animal by retroviral gene delivery can be enhanced by partial hepatectomy, and that the retrovirus-mediated FVIII-B cDNA delivery to regenerating liver may be an alternative method for the expression of FVIII-B cDNA in vivo. PMID- 10391102 TI - Development of erythrocytosis in the course of essential thrombocythemia. AB - Erythrocytosis is not a feature of essential thrombocythemia (ET); this is the most important difference between ET and polycythemia vera (PV). Transformation of ET to PV has only rarely been described. We have reviewed the blood cell counts of 170 ET patients with a median follow-up of 63 months (range 11-313). Eleven of 170 patients (6.5%) developed erythrocytosis at a median of 29 months (range 12-138) after the diagnosis of ET. According to the present results, the development of erythrocytosis in patients with ET is not a rare phenomenon. PMID- 10391103 TI - Human acute myeloblastic leukemia-ascites model using the human GM-CSF- and IL-3 releasing transgenic SCID mice. AB - To generate an appropriate model for human acute myeloblastic leukemia (AML), we have successfully established a human hematopoietic growth factor-dependent AML cell line (TF-1 and UT-7/GM)-ascites model using human granulocyte-macrophage colony-stimulating factor (hGM-CSF)- and human interleukin 3 (hIL-3)-releasing transgenic (Tg)-SCID mice. When 1 x 10(7) cells of TF-1, a human erythroleukemia cell line, were transplanted into the peritoneum of irradiated Tg-SCID mice (TF-1 ip/Tg-SCID mice), TF-1 cells grew in both the single cell suspension form (asTF 1) and solid form in ascites and invaded various tissues: lungs, liver, pancreas, and genitals, 3-6 weeks following transplantation. Subsequently, 0.5-1 x 10(7) cells of UT-7/GM, a subline of the UT-7 human megakaryoblastic leukemia cell line, grown in the back of hGM-CSF Tg-SCID mice after subcutaneous inoculation, were transplanted into the peritoneum of other irradiated hGM-CSF Tg-SCID mice. After 4 weeks, UT-7/GM cells (asUT-7/GM) also grew in the same manner as TF-1 cells in hGM-CSF Tg-SCID mice. Analysis of the cells from the peritoneum and tissues by PCR amplifying ALU and human GM-CSF receptor beta sequences and by immunohistochemical staining using anti-human CD45 revealed that they possessed the original characteristics of the parental cells. To confirm the usefulness of this human AML-ascites model, experimental treatment of AML cells grown in these mice was carried out with a differentiation inducer, delta-aminolevulinic acid (deltaALA), which induces hemoglobin synthesis for TF-1 in vitro and is thus regarded as an anti-leukemia drug candidate. Unexpectedly, growth promotion of TF 1 cells was observed in the treated TF-1 ip/hIL-3 Tg-SCID mice without differentiation to erythroid cells after treatment with delta-ALA (5 mM) for 7 days. These results indicate that Tg-SCID mice can support the growth of human hematopoietic growth factor-dependent AML cell lines which are usually rejected by SCID mice, without modification of the parental cell characteristics. In addition, this Tg-SCID leukemia-ascites model may become a useful preclinical tool for estimation of drug efficacy in vivo, since the drug candidate which was promising in vitro did not act in the same manner in vivo. PMID- 10391104 TI - Why is acute leukemia more common in males? A possible sex-determined risk linked to the ABO blood group genes. AB - Acute leukemia is more common in males at almost every age, and this fact remains unexplained. A study was carried out in northeast peninsular Malaysia, where the population is predominantly Malay, to examine whether there was a difference in ABO blood group distribution between males and females with acute leukemia (AL). The ABO blood groups of 109 male and 79 female patients with AL (98 ALL, 90 AML) were compared with those of 1019 controls. In the control population, 39.7% were group O. Among males with AL, 39.4% were group O, whereas among females with AL, the proportion was 24.1% (p=0.03). The same trend to a lower proportion of group O among females was seen if the group was divided into adult/pediatric or lymphoblastic/myeloblastic groups, though these differences were not statistically significant. If these findings can be confirmed, they suggest the presence of a "sex-responsive" gene near to the ABO gene locus on chromosome 9, which relatively protects group O women against AL, at least in our population. The existence of such a gene might also partly explain why acute leukemia, and possibly other childhood cancers, are more common in males. PMID- 10391105 TI - Acute myeloid leukemia M2 and t(8;21)(q22;q22) with an unusual phenotype: myeloperoxidase (+), CD13 (-), CD14 (-), and CD33(-). AB - Cases of myeloid surface antigen-negative acute myeloid leukemia (AML) are rare. We describe the morphological, cytochemical, immunologic, and cytogenetic features of two patients with AML with maturation (FAB M2) and the phenotype MPO+, CD13 (-), CD33(-), CD56(+). Cytogenetic studies demonstrated t(8;21)(q22;q22). These findings suggest an association between the lack of CD13 and CD33 in myeloperoxidase-positive AML and the presence of t(8;21). PMID- 10391106 TI - Cryptic Mucor infection leading to massive cerebral infarction at initiation of antileukemic chemotherapy. AB - A 74-year-old man with newly diagnosed acute myelogenous leukemia unexpectedly suffered a massive cerebral infarct on day 2 of induction chemotherapy. Clinically, the hemorrhagic infarct was thought to be due to leukostasis and thrombocytopenia. Necropsy, however, revealed that Zygomycetes-type hyphae had infiltrated cerebral vessels in and near the infarct. The fungal infection was clinically silent otherwise, although fungal elements were also identified in the lung at autopsy. This case illustrates how closely fungal infection may resemble a leukemia-associated cerebrovascular accident. PMID- 10391107 TI - Prevention of hepatitis B flare-up during chemotherapy using lamivudine: case report and review of the literature. AB - Reactivation of chronic hepatitis B in patients receiving cytotoxic treatment for non-Hodgkin's lymphoma is well documented. We report a case of a patient with chronic hepatitis B who was treated by chemotherapy because of non-Hodgkin's lymphoma. After the second cycle of chemotherapy she developed a severe flare-up of hepatitis B. Liver biopsy revealed highly active hepatitis and confluent necroses. Within 3 weeks, the patient recovered spontaneously. Prophylactic treatment with lamivudine (Epivir,Glaxo-Wellcome, 150 mg b.i.d.) led to a decrease of HBV-DNA below the detection limit. Further chemotherapy was administered and autologous stem cell transplantation was successfully performed without another reactivation of hepatitis B. Antiviral treatment was stopped 16 weeks after stem cell retransfusion. So far, no further flare-up of hepatitis B has occurred and the patient's lymphoma has not relapsed. Thus, the case described here indicates a possible role of lamivudine in preventing hepatitis B flare-up during antineoplastic chemotherapy. We suggest that lamivudine be considered for prophylaxis against fulminant hepatitis in patients with chronic HBV infection undergoing high-dose antineoplastic therapy. PMID- 10391108 TI - New Insights into Disintegrin Metalloproteases. PMID- 10391109 TI - Innate Immunity: The Bridge between Adaptive Immunity and Inflammation, December 17, 1998, New York Academy of Sciences, New York, NY, USA. PMID- 10391110 TI - Risks of anti-inflammatory drug-associated damage. AB - Non-steroidal anti-inflammatory drug toxicity to the upper gastrointestinal tract differs consistently and strongly between the individual agents. Exact measures of the difference are not yet available because meta-analyses combining individual data sets from a sufficiently large number of subjects and incorporating dosage information are not available. The importance of obtaining such information is emphasised by simple modelling based upon expected usage patterns and disease risks. The adverse effects of the non-steroidal anti inflammatory drugs (NSAIDs) are, in terms of general population impact, the most common and important of all adverse effects of pharmaceuticals in any group. This is because they are widely used, and the adverse effects are common. Amongst these adverse effects, gastrointestinal problems, manifested by peptic ulceration and its complications, are by far the most frequent. Approaches to prevention can be of three general types. The first is the characterisation and mitigation of the circumstances under which the effects occur, the second the use of preventative treatments, and the third the introduction of selective agents by which therapeutic properties are retained and adverse effects minimised. This commentary concentrates on characterising the extent of risks. However, a good understanding will not be achieved without knowledge of the general epidemiology of peptic ulceration. PMID- 10391111 TI - Regulation of lymphocyte traffic by adhesion molecules. AB - Lymphocytes are antigen specific cells whose effector function is acquired through complex differentiation pathways. This implies, firstly, antigen encounter and recognition at specific sites, and, subsequently, the transition from a naive to a memory/effector phenotype. Clonotypically expanded cells must then be capable of recirculating to the tissue where their effector function is needed. To this aim, defined receptor-counter receptor pairs are expressed on lymphocytes versus endothelial cells. Extravasation is therefore a key-process in this scenario. Indeed, different lymphocyte subsets display distinct recirculation patterns and capability to migrate into lymphoid and non-lymphoid tissues. As a general rule, naive lymphocytes preferentially migrate into secondary lymphoid organs, where all the requirements for effective antigen presentation and differentiation are available; in contrast, memory/effector lymphocytes preferentially migrate to peripheral tissues, such as skin and mucosa. We review here the molecular events that regulate leukocyte extravasation and the specific migration properties acquired by both naive and memory/effector lymphocytes under physiological and pathological conditions. PMID- 10391112 TI - Clinical experience with cyclooxygenase-2 inhibitors. AB - Increasing amounts of experimental and clinical data support the role of selective cyclooxygenase (COX)-2 inhibition in anti-inflammatory processes and the role of COX-1 inhibition in increasing the frequency of side effects. This article reviews the regulation of COX-2 in inflammatory processes based on in vitro and in vivo work. In addition, it summarizes the various in vitro assays used to classify the new generation of selective and highly selective inhibitors of COX-2, since prior categorization of NSAIDs does not satisfactorily encompass the COX-2 concept. Finally, the latest published clinical data of new selective and highly selective inhibitors of COX-2 (meloxicam, nimesulide, etodolac, celecoxib and MK966) are discussed. PMID- 10391113 TI - The roles of interleukin-6 and interleukin-10 in B cell hyperactivity in systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is an autoimmune disease most prevalent in women between the ages of twenty and sixty. Successful treatment remains challenging due to a lack of understanding of the underlying mechanisms and multiple symptoms ranging from skin rashes to glomerulonephritis. The pathogenesis of SLE has been linked to a B-cell hyperproliferation unique to afflicted patients. These B-cells generate large quantities of IgG autoantibodies, ultimately capable of leading to lupus nephritis and renal failure. The significance of cytokines in SLE and in murine lupus, a related disease in mice, has been debated, particularly with respect to B-cell activity. Potential roles of auto-regulatory and inflammatory cytokines have been investigated. In particular, IL-6 and IL-10 have been shown to be key factors in regulating autoantibody-secreting B-cell activity in lupus. Here, we will provide a critical overview of our current knowledge of the regulatory roles of these two cytokines in SLE. PMID- 10391114 TI - The effect of captopril on histamine release from purified rat mast cells. AB - OBJECTIVE: Captopril as inhibitor of angiotensin-converting enzyme is widely used in cardiovascular therapy, however, in some patients this drug causes allergic side effects. This fact suggests that captopril may release histamine from mast cells. MATERIALS AND METHODS: Peritoneal mast cells were obtained from rats. The cells were purified by Percoll. Aliquots of mast cells were incubated with captopril or with vehicle. Histamine was determined by the spectrofluorimetric assay. RESULTS: The results were analyzed with the Kruskall-Wallis (ANOVA) test. The study has shown that captopril releases histamine from mast cells. The process depends on Ca2+ presence in the incubation environment. Sodium cromoglycate, as mast cell membrane stabilizer, inhibits the effect of captopril. CONCLUSIONS: Our results suggest that allergic side effects of captopril may be linked to histamine release from mast cells. PMID- 10391115 TI - Exposure of intestinal epithelial cell HT29 to bile acids and ammonia enhances Mac-1-mediated neutrophil adhesion. AB - OBJECTIVE: Bile acids and ammonia (NH3/NH4+) are cytotoxic metabolic products, which are known to influence intestinal epithelial functions such as colonic chloride secretion. In this study, we have investigated the effect of bile acids and ammonia on neutrophil-intestinal epithelial adhesive interaction. MATERIALS AND METHODS: Confluently cultured HT29 cells were treated with bile acids or ammonium chloride. Then, 51Cr-labeled neutrophils were added to HT29 cell monolayers, and neutrophil adhesion was assessed by gamma-scintillation counting. RESULTS: Treatment of HT29 cells with 0.1 mM CDCA (chenodeoxycholic acid), DCA (deoxycholic acid), CA (cholic acid) or ammonium chloride but not UDCA (ursodeoxycholic acid) for 30 min increased neutrophil adhesion about 4-folds (p<0.01). The increased adhesion was inhibited 82-91% by 10 microg/ml anti-CD11b and anti-CD18 mAbs (p<0.01), but not by anti-CD11a and anti-CD54 (ICAM-1) mAbs. Interestingly, flow cytometric analysis revealed that ICAM-1 expression on HT29 cells was not changed by bile acid- or ammonia-treatment. In addition, the increased adhesion was inhibited about 65 % by proteinase K-treatment (10 microg/ml, 1 min, p<0.05) but not cycloheximide-treatment (1 microg/ml, 30 min) of HT29 cells. CONCLUSIONS: These observations indicate that exposure of HT29 cells to bile acids or ammonia induces CD11b/CD18 (Mac-1) dependent- but CD11a/CD18 (LFA-1) independent-neutrophil adhesion to intestinal epithelial cells, and ICAM-1 is unlikely involved in the interactions. Furthermore, epithelial ligand(s) for neutrophils are protein molecule(s) which are expressed on the cell surface independent of protein synthesis. PMID- 10391116 TI - Functional neurokinin NK-1 receptor expression in rat peritoneal mast cells. AB - OBJECTIVE AND DESIGN: Recently, Ogawa et al. [17] reported that the peritoneal mast cells (PMCs) of rats can release histamine by substance P (SP) in a receptor dependent manner. In the present study, we confirmed and extended their findings. MATERIAL: PMCs were isolated from six strains of rats. In some experiments, peritoneal cells in the non-MC fraction were used. METHODS: PMCs were incubated with SP, neurokinin (NK) receptor agonists or antagonists, and histamine content in the supernatant was measured. In the binding assay, PMCs were incubated with [125I]BH-SP together with SP or NK receptor antagonists. NK-1 receptor mRNA was detected using a reverse transcription-polymerase chain reaction (RT-PCR) assay. RESULTS: PMCs from Slc: Wistar and F344/NSlc were highly sensitive to SP, leading to histamine release, whereas those from Slc:SD and three other strains were not. PMCs from Slc:Wistar and F344/NSlc also released histamine in the presence of an NK-1 agonist. The histamine release induced by SP and the NK-1 agonist was inhibited by the NK-1 receptor antagonists, FK888 and CP-99,994. [125I]BH-SP binding experiments revealed that PMCs from Slc:Wistar rats possessed a single high affinity binding site for SP and that the binding was blocked by NK-1 receptor antagonists. Peritoneal cells in the non-MC fraction exhibited no appreciable binding. In the RT-PCR assay, expression of NK-1 receptor mRNA was evident in Slc:Wistar PMCs, but not in the non-MC fraction from Slc:Wistar or Slc: SD PMCs. CONCLUSION: These data demonstrate the existence of functional NK-1 receptors on freshly isolated PMCs in at least some strains of rats. PMID- 10391117 TI - Cartilage proteoglycan degradation by a mouse transformed macrophage cell line is mediated by macrophage metalloelastase. AB - OBJECTIVE AND DESIGN: Identify and characterize the matrix metalloproteinase responsible for cartilage proteoglycan degradation mediated by a macrophage cell line in a cell culture model that resembles some aspects of rheumatoid pannus. MATERIALS OR SUBJECTS: Supernatants from the transformed mouse macrophage cell line J774A.1 were used to purify the proteoglycan degrading activity. METHODS: J774A.1 macrophage culture supernatants were purified by sequential column chromatography and proteins were identified by zymography, western blotting and amino acid sequence analysis. Cartilage degradation was measured using 35S labeled bovine nasal cartilage. RESULTS: The cartilage degrading proteolytic activity in the mouse macrophage supernatants proved to be due to two major proteins with approximate molecular masses of 48 kDa and 22 kDa that were identified as macrophage metalloelastase (MME). Incubation of purified MME at 37 degrees C for up to 16 h resulted in the processing of the 48 kDa protein to several novel bands including a previously undescribed protein of approximately 25 kDa without accumulation of fully processed 22 kDa protein. A number of proteinases increased the rate of this processing. J774A.1 macrophage metalloelastase degraded cartilage proteoglycan with an efficiency approximately equal to human macrophage metalloelastase (MMP-12) and matrilysin (MMP-7) and twice that of stromelysin-1 (MMP-3). CONCLUSIONS: These data identify the cartilage proteoglycan degrading metalloproteinase secreted by J774A.1 macrophages in this cell culture model as MME, and describes mechanisms of activation and processing of this enzyme that may play an important role in cartilage degradation. PMID- 10391119 TI - Studies on purine enzymes in experimental colitis. AB - Although the role of adenosine deaminase (ADA), adenylate deaminase (AMP-DA), purine nucleoside phosphorylase (PNP) is well documented in gastric and intestinal carcinoma, their role in inflammatory bowel diseases remains unknown. In the present study, we investigated the profile of these enzymes in blood and intestinal tissues during colitis. Colitis induced in Wistar rats by acetic acid was monitored by a marker enzyme myeloperoxidase (MPO). The tissue levels of MPO increased on 1, 2, 5 and 6 days post-administration (PA) of acetic acid and declined to the control levels by day 7 PA. In parallel the blood levels of ADA and AMP-DA decreased on days 1, 2 and 5 without any significant change on days 6 and 7 PA. Similar observations were recorded for these enzymes in the cytosolic extracts of colonic tissue specimens. In contrast, PNP remained unaltered in both blood and tissue samples. These findings suggest an inverse-relationship between inflammation and purine deaminases in both blood and tissues. PMID- 10391120 TI - Comparative binding study of rat natriuretic peptide receptor-A. AB - The natriuretic peptide receptor-A (NPR-A) is involved in blood pressure and body fluid regulation in order to help maintain cardiovascular homeostasis. It has been shown that these biological effects are mediated through the natriuretic peptide family of hormones, which bind NPR-A according to the rank order ANP>BNP>>CNP. Previous studies performed with rat kidney papillary tissue suggested the existence of an heterologous NPR-A population since two binding components were obtained for pBNP32, one of high affinity (pK 9.4 +/- 0.1) and the other of lower affinity (pK 7.5 +/- 0.1), while in the same preparation rANP28 binding displayed the expected affinity (pK 10.22 +/- 0.01) and was best fitted with a model involving a single class of binding sites. This apparent heterogeneity of NPR-A in rat kidney papillae could be explained by the presence of two receptor isoforms or of monomeric and oligomeric forms of the same receptor. To investigate the NPR-A binding heterogeneity, we have cloned the rat NPR-A from PC12 cells and compared its pharmacological profile with that of the papillae. Our results with rat NPR-A transfected Cos-P cells show an equivalent pharmacological profile as with the rat tissue, i.e. a high affinity for rANP28 (pK 10.4 +/- 0.1) and two distinctive affinities for pBNP32 (pK 9.74 +/- 0.05 and 7.8 +/- 0.1). Although multiple receptor glycoforms were sometimes detectable by western blotting, only one molecular form was obtained by cross-linking with 125I rANP28. It thus appears that NPR-A alone can account for the two binding components found in the rat papillae and that a single molecular form of the protein is implicated. We therefore propose that the oligomerization state of the receptors could be responsible for the apparent binding heterogeneity of rat NPR A. PMID- 10391118 TI - Mitochondrial involvement in bladder function and dysfunction. AB - Benign bladder pathology resulting from prostatic hypertrophy or other causes is a significant problem associated with ageing in humans. This condition is characterized by increased bladder mass, decreased urinary flow rate, decreased compliance, and these and other changes in bladder function often subject patients to increased risk of urinary tract infection. While the physiologic attributes of benign bladder pathology have been extensively described in humans and in various animal model systems, the biochemical and molecular genetic bases for that pathology have only recently been investigated in detail. Studies demonstrate that mitochondrial energy production and utilization are severely impaired in bladder smooth muscle during benign bladder disease, and to a large extent this realization has provided a rational basis for understanding the characteristic alterations in urinary flow and compliance in bladder tissue. Recent investigations targeting the detailed molecular basis for impaired mitochondrial function in the disease have shown that performance of the organellar genetic system, and to a large extent that of relevant portions of the nuclear genetic system as well, is severely aberrant in bladder tissue. In this article, we discuss the physiologic aspects of benign bladder disease, summarize biochemical evidence for the altered mitochondrial energy metabolism that appears to underlie bladder pathology, review the structure and function of the mitochondrial genetic system, and discuss molecular genetic studies of that system which have begun to provide a mechanistic explanation for the biochemical and physiological abnormalities that characterize the disease. We also discuss areas for further research which will be critically important in increasing our understanding of the detailed causes of benign bladder pathology. PMID- 10391122 TI - Protection against UVB inactivation (in vitro) of rat lens enzymes by natural antioxidants. AB - Oxidative damage, through increased production of free radicals, is believed to be involved in UV-induced cataractogenesis (eye lens opacification). The possibility of UVB radiation causing damage to important lenticular enzymes was assessed by irradiating 3 months old rat lenses (in RPMI-1640 medium) at 300 nm (100 microWcm(-2)) for 24 h, in the absence and presence of ascorbic acid, alpha tocopherol acetate and beta-carotene. UVB irradiation resulted in decreased activities of hexokinase, glucose-6-phosphate dehydrogenase, aldose reductase, and Na, K- ATPase by 42, 40, 44 and 57% respectively. While endopeptidase activity (229%) and lipid peroxidation (156%) were increased, isocitrate dehydrogenase activity was not altered on irradiation. In the presence of externally added ascorbic acid, tocopherol and beta-carotene (separately) to the medium, the changes in enzyme activities (except endopeptidase) and increased lipid peroxidation, due to UVB exposure, were prevented. These results suggest that UVB radiation exerts oxidative damage on lens enzymes and antioxidants were protective against this damage. PMID- 10391121 TI - Cardiac myofibrillar proteins: biochemical markers to estimate myocardial injury. AB - Ischaemic heart disease represents the most common of the serious health problems in the contemporary society and acute myocardial infarction (AMI) is the major cause of cardiovascular morbidity and death. The accurate localization and determination of the infarct size and the volume of myocardium at risk at the time of insult is crucial and vital for the choice of treatment. Initially the ischaemic cells are reversibly injured. However, if these changes are not reverted at the earliest, it results in the death of the myocyte. This irreversible myocyte necrosis travels transmurally towards epicardium in the form of a wavefront. A timely intervention during evolving infarct could reduce and delimit the infarct and preserve the left ventricular function. Enzyme analysis and electrocardiography (ECG) along with the clinical history of the patient is still considered to constitute a reliable triad in the diagnosis of myocardial infarction (MI). Efforts have been made to relate infarct size with the serum enzyme level changes without much success. In addition, a number of specialist techniques such as planar radioisotope imaging, single photon emission computed tomography (SPECT), positron emission tomography (PET), Echocardiography, Ventriculography and nuclear magnetic resonance (NMR) imaging have been devised to support diagnosis in the patients who show ambiguous symptoms and ECG findings. However most of these procedures are unavailable to the patients due to economic reasons while others have suffered due to non-availability of ideal radiopharmaceuticals. Major advances have been made in the methods based on immunological techniques to improve the detection and estimation of infarct. These methods are exclusively based upon the production and availability of specific antibodies against intracellular, cardiac specific components. PMID- 10391123 TI - IgG, IgG1 and IgM response in Trichinella spiralis-infected mice treated with 4 deoxypirydoxine or fed a Vitamin B6-deficient diet. AB - The aim of this study was to investigate the effect of pyridoxine (Vitamin B6) deficiency on the immunological response of BALB/c mice infected with the parasite T. spiralis. Specific anti-parasite IgM and IgG immunoglobulins were detected by ELISA method in the serum of treated animals at different periods for 60 days post infection. Vitamin B6-deficiency was induced in two separate groups of mice by either (1) maintaining the mice on a Vitamin B6-deficient synthetic pellet diet for 40 days before infection, or (2) by daily intraperitoneal injection of 8 x 10(5) M/100 microl of 4-Deoxypyridoxine (4-DPD), a potent antagonist of Vitamin B6 for 20 days prior to infection. These two groups of mice were then injected with 100 larvae (L1-T. spiralis) per os. Parasite burdens in the mice were observed by light microscopy. Cysts were present in the diaphragms of the mice after 60 days post-infection. Parasite specific IgG, as well as IgG1 levels were determined in the sera of infected mice fed a normal diet. These levels were found to be lower in the 4-DPD-treated mice compared to the untreated mice. The inhibition started from the 10th day and continued to the 60th day, and in the 4-DPD-treated group the inhibition initiated after 24 h to 60 days. IgM level also was depressed by 4-DPD, starting from 24 h after injection of the compound. In mice fed Vitamin B6-deficient diets the levels of IgG were lower than in mice fed normal diets. These results show that BALB/c mice infected with T. spiralis and fed either a Vitamin B6-deficient diet or a diet which included the Vitamin B6-antagonist, 4-DPD, both influence the course of IgG, IgG1 and IgM production. PMID- 10391124 TI - Interaction of cholera toxin and Escherichia coli heat-labile enterotoxin with glycoconjugates from rabbit intestinal brush border membranes: relationship with ABH blood group determinants. AB - The capacity of cholera toxin (CT) and type I heat-labile enterotoxin produced by Escherichia coli isolated from human intestine (LTh) to interact with glycoconjugates bearing ABH blood group determinants from rabbit intestinal brush border membranes (BBM) was studied. On the basis of the type of intestinal compounds related to the human ABH blood group antigens, rabbits were classified as AB or H. Toxin binding to the intestinal glycolipids and glycoproteins depends on the blood group determinant borne by the glycoconjugate and on the analyzed toxin. LTh was capable of interacting preferentially with several blood group A- and B-active BBM glycolipids compared to those isolated from animals lacking these antigens (H rabbits). Also, LTh preferably bound to several BBM glycoproteins from AB rabbit intestines compared to those from H ones. One of these glycoproteins, the sucrase-isomaltase complex (EC 3.2.1.48-10) isolated from AB and H rabbits showed the same differential LTh binding. Conversely, CT practically did not recognize either blood group A-, B-, or H-active glycolipids and glycoproteins. These results may be relevant for carrying out in vivo experiments in rabbits in order to disclose the role of ABH active glycoconjugates in the secretory response induced by LTh in rabbit intestine. PMID- 10391125 TI - The role of hydroxyl radical as a messenger in the activation of nuclear transcription factor NF-kappaB. AB - Although it is generally believed that reactive oxygen species activate NF kappaB, a primary oxidative stress-responsive transcription factor, it is unclear which one among these species causes NF-kappaB activation. Our hypothesis is that hydroxyl radical (*OH) functions as a messenger for the activation of NF-kappaB. Jurkat cells, macrophages and JB6 cells were used to test this hypothesis. Cr(VI), silica and ZnO were used as sources of *OH radicals. None of these *OH generating systems involves exogenous H2O2. Cr(VI) expressed enhanced activity in induction of NF-kappaB in Jurkat cells. This activation of NF-kappaB was decreased by a metal chelator, diethylene triaminepentaacetic acid or a H2O2 scavenger, catalase, but was increased by superoxide dismutase. Mn(II), which reacts with Cr(IV) to inhibit this metal ion-mediated *OH generation, decreased the NF-kappaB activation. Sodium formate, an *OH radical scavenger, also inhibited the NF-kappaB activation. Electron spin resonance measurements show that Cr(VI) was reduced by Jurket cells to Cr(IV) and Cr(V). During the reduction process, molecular oxygen was reduced to O2 and then to H2O2, which reacted with Cr(IV) and Cr(V) to generate *OH radical. The *OH generation correlated with the Cr(VI)-induced NF-kappaB activation. Similarly, silica caused NF-kappaB activation in macrophages via the *OH radical-mediated reaction. This radical was generated via metal mediated reaction from H2O2, which was generated by the reduction of molecular oxygen via O2- as an intermediate during the silica stimulated 'respirable burst'. Silica particles did not cause *OH generation either in Jurket or in JB6 cells and thus did not cause any observable NF-kappaB activation in these cells. ZnO induced NF-kappaB activation in JB6 cells through the generation of *OH resulting from light irradiation of ZnO which was measured by electron spin resonance. The results thus show that *OH radical functions as a messenger for NF-kappaB activation. Antioxidants, which scavenge *OH radical or its precursors, inhibit NF-kappaB activation. Metal chelators, which make metal ions incapable of generating *OH from H2O2, inhibit activation of this transcription factor. PMID- 10391126 TI - Three different forms of hexokinase are identified during Tuber borchii mycelium growth. AB - Truffles are ectomycorrhizal fungi which have a great dependence on carbohydrates supplied by their host plants. The catabolism of hexoses in the mycobiont is important for the production of energy, and the first enzyme in the hexose assimilation pathways is hexokinase. This study reports differences in the expression of this enzyme during the growth of Tuber borchii Vittad. mycelium (strain ATCC 96540). Three hexokinase activities (HKM1, HKM2 and HKM3) were isolated by anion-exchange chromatography and partially purified. HKM1 and HKM2 were present in the linear phase at 15-50 days of growth. Two remarkable differences were found in the sugar-phosphorylating activity and stability of HKM1 and HKM2. HKM2 did not phosphorylate the fructose and it was present in the chromatographic profile only when substrates such as glucose, glucosamine or mannose were added to the extraction buffer. On the contrary, HKM1 utilized also fructose and was detected under all the experimental conditions used. HKM3 was the only molecular form observed after 70 days, when the fungus growth had reached a plateau. To our knowledge these results represent the first evidence for the presence in T. borchii mycelium of three distinct enzymatic forms of hexokinase which are differently expressed during growth of the fungus. PMID- 10391127 TI - Epigenetic inhibition of lysyl oxidase transcription after transformation by ras oncogene. AB - Lysyl oxidase is an extracellular enzyme involved in connective tissue maturation that also acts as a phenotypic suppressor of the ras oncogene. To understand how this suppressor is controlled, gene transcription was studied and the promoter was characterized. Nuclear runoff transcription assays indicated that the markedly reduced amounts of lysyl oxidase message detected after ras transformation resulted from inhibition of lysyl oxidase transcription. Interferon-mediated phenotypic reversion of ras transformed cells, in which the ras oncogene continued to be expressed, was accompanied by the restoration of lysyl oxidase transcription. Reporter gene assay of a transfected mouse lysyl oxidase promoter indicated that it was active in the transformed background, despite the silencing of the endogenous lysyl oxidase promoter. The detection of comparable amounts of mRNA for transcription factors IRF-1 and IRF-2 in normal and ras-transformed cell lines suggests that the differential transcription of lysyl oxidase was not due to regulation of IRFs. Lysyl oxidase promoter activity was localized to a 126 bp region that includes two consensus TATA boxes with associated confirmed cap signals. Analysis of a human lysyl oxidase promoter sequence indicated similar promoter elements and extensive sequence identity with the mouse promoter. The binding of transcription factor AP2 to sites predicted in the control region was confirmed by DNase footprinting. Lysyl oxidase transcription was stimulated by dexamethasone treatment of cells, but this effect could not be assigned within the approximately 3 kb region tested in reporter gene constructs. The promoter activity of the lysyl oxidase reporter gene construct was completely abolished by in vitro DNA methylation, suggesting that the transcriptional suppression after transformation by the ras oncogene may involve DNA methylation. PMID- 10391128 TI - Anabolic effect of daidzein on cortical bone in tissue culture: comparison with genistein effect. AB - The effect of daidzein on cortical bone in vitro was investigated. Femoral diaphyseal tissues obtained from elderly female rats were cultured for 24 h in Dulbecco's modified Eagle's medium (high glucose, 4.5%) supplementation with antibiotics and bovine serum albumin. The experimental cultures contained 10(-7) to 10(-5) M daidzein. The presence of daidzein (10(-6) and 10(-5) M) caused a significant increase of alkaline phosphatase activity, deoxyribonucleic acid (DNA) and calcium contents in bone tissues. This effect was equal to that of genistein (10(-6) and 10(-5) M). Daidzein (10(-5) M) or genistein (10(-5) M) induced increase of calcium content and alkaline phosphatase activity in bone tissues was completely prevented by cycloheximide (10(-6) M), an inhibitor of protein synthesis. Anabolic effect of daidzein and genistein on bone components was equal to that of 17beta-estradiol (10(-8) M). The effect of isoflavohoids was not enhanced by the addition of 17beta-estradiol. The combination of daidzein and genistein did not have an additive effect. These findings indicate that daidzein has an anabolic effect on bone metabolism in tissue culture in vitro, and that this effect is equal to genistein effect. Isoflavonoids may stimulate bone formation and mineralization. PMID- 10391129 TI - Aging differentially modulates the expression of collagen and collagenase in dermal fibroblasts. AB - This study was conducted to investigate the effects of aging on collagen and collagenase expression by human dermal fibroblasts. To evaluate this effect, the expression of these ECM was determined and compared between either fetal and adult fibroblasts or dermal fibroblasts at various passages. A total of 13 cell strains, 8 fetal foreskin and 5 adult dermal fibroblasts, were grown to 80-90% confluency and their rates of cell proliferation and expression of mRNA for collagenase (MMP-1) and pro alpha1(I) chain of type I collagen was determined and compared. Fetal cells had a significantly higher rate of proliferation relative to adult fibroblasts evaluated within 10 days of culture. Northern analysis was used to evaluate the steady state levels of mRNA in these cells. The result of these experiments revealed a significantly greater expression of mRNA for collagenase (58.6 +/- 7.7 vs. 9.9 +/- 1.5, p < 0.05) in strains of adult fibroblasts. This was consistent with collagenase activity of conditioned medium derived from adult cells relative to fetal fibroblasts. However the expression of pro alpha1 (I) chain of type I collagen mRNA was not significantly (56.2 +/- 5.2 vs. 58.5 +/- 3.5) different between adult and fetal fibroblasts. This finding was confirmed by measuring total collagen production present in conditioned medium of these cells using hydroxyproline as an index for collagen production. The cellular response to IGF-1 and IFN-alpha2b as representatives of fibrogenic and anti-fibrogenic factors were also evaluated. When expression of collagenase was used as an indication for cellular response, the degree of this response to IGF-1 but not IFN-alpha2b was significantly greater in fetal relative to adult cells. Serial passage was also used as an in vitro model for aging fibroblasts and found a gradual reduction in pro alpha1(I) chain of type I collagen mRNA and hydroxyproline formation due to passaging. In conclusion, a slower rate of proliferation, a greater collagenase activity and expression of collagenase mRNA by aging fibroblasts could be some of the main reasons for attenuation of wound healing in elderly patients. PMID- 10391130 TI - Mechanism of stimulation of glucose transport in response to inhibition of oxidative phosphorylation: analysis with myc-tagged Glut1. AB - To determine the role of 'translocation' vs. 'activation' of Glut1 in the stimulation of glucose transport in response to inhibition of oxidative phosphorylation, we measured the abundance of myc-tagged Glut1 in plasma membrane of stably transfected Clone 9 cells, a rat liver cell line expressing only the Glut1 isoform. The myc epitope-tag is located between Ile56 and Pro57 in the putative first extracellular loop of Glut1. Under basal conditions, transfected cells expressed approximately 3 fold higher levels of Glut1 and exhibited a approximately 3 fold higher rate of glucose transport than non-transfected cells. To delineate the mechanism mediating the stimulation of glucose transport by a azide we employed two strategies: (1) mild cell surface biotinylation followed by isolation of plasma membranes and quantitation of Glut1 sites in Western blots employing anti-Glut1 and anti-myc antibodies, and (2) quantitative immunofluorescence of myc epitopes in plasma membrane sheets. The rate of glucose transport increased 2.9 +/- 0.5 fold in transfected cells exposed to 5 mM azide for 1 h. Exposure to azide, however, resulted in no significant increase in Glut1 content of plasma membranes using anti-Glut1 or anti-myc antibodies in Western blots (1.0 +/- 0.1 and 0.9 +/- 0.2 fold, respectively; azide/control), and was associated with no detectable increase in immunofluorescence using either anti Glut1 or anti-myc antibodies (p > 0.1 for both measurements). Treatment of cells with cobalt chloride (employed as a positive control) resulted in marked increases in glucose transport, cell and plasma membrane Glut1 content, and immunofluorescence of plasma membrane sheets (8-10 fold increase in each parameter). We conclude that the stimulation of glucose transport by azide results mainly from activation of Glut1 transporters pre-existing in the plasma membrane. PMID- 10391131 TI - Molecular cloning and developmental expression of rat glycogenin in cardiac tissue. AB - Glycogenin is a self-glycosylating protein required to initiate glycogen biosynthesis. Utilizing the differential display technique to analyze changes in gene expression during early postnatal cardiac development, we have isolated and cloned a 484 bp cDNA fragment that corresponds to the 3' end of rat glycogenin. Northern blot analysis on neonatal cardiac tissues demonstrated hybridization to a 1.7-1.8 kb transcript, which was highly expressed at 3 days and at progressively reduced levels at 1, 2, 3 and 4 weeks of age. A 1624 bp fragment of rat glycogenin was cloned by RT-PCR that includes a 1002 bp open reading frame encoding a 333 amino acid protein. At the nucleotide level, rat glycogenin exhibited 87.2 and 83.6% identity with human and rabbit glycogenin over the open reading frame. The deduced amino acid sequence showed 86.7 and 83.4% identity with human and rabbit sequences, respectively. Given the significance of glycogenin in glycogen biosynthesis, the results of this study suggest a possible molecular basis for the regulation of glycogen during early postnatal cardiac development. In addition, the nucleotide and amino acid sequences of rat glycogenin may be used to investigate the physiological and pathophysiological roles of glycogenin in rat tissues. PMID- 10391133 TI - Inhibition of glucose-induced insulin release by 3-O-methyl-D-glucose: enzymatic, metabolic and cationic determinants. AB - The analog of D-glucose, 3-O-methyl-D-glucose, is thought to delay the equilibration of D-glucose concentration across the plasma membrane of pancreatic islet B-cells, but not to exert any marked inhibitory action upon the late phase of glucose-stimulated insulin release. In this study, however, 3-O-methyl-D glucose, when tested in high concentrations (30-80 mM) was found to cause a rapid, sustained and not rapidly reversible inhibition of glucose-induced insulin release in rat pancreatic islets. In relative terms, the inhibitory action of 3-O methyl-D-glucose was more marked at low than high concentrations of D-glucose. It could not be attributed to hyperosmolarity and appeared specific for the insulinotropic action of D-glucose, as distinct from non-glucidic nutrient secretagogues. Although 3-O-methyl-D-glucose and D-glucose failed to exert any reciprocal effect upon the steady-state value for the net uptake of these monosaccharides by the islets, the glucose analog inhibited D-[5-3H]glucose utilization and D-[U-14C]glucose oxidation. This coincided with increased 86Rb outflow and decreased 45Ca outflow from prelabelled islets, as well as decreased 45Ca net uptake. A preferential effect of 3-O-methyl-D-glucose upon the first phase of glucose-stimulated insulin release was judged compatible with an altered initial rate of D-glucose entry into islet B-cells. The long-term inhibitory action of the glucose analog upon the metabolic and secretory response to D glucose, however, may be due, in part at least, to an impaired rate of D-glucose phosphorylation. The phosphorylation of the hexose by beef heart hexokinase and human B-cell glucokinase, as well as by parotid and islet homogenates, was indeed inhibited by 3-O-methyl-D-glucose. The relationship between insulin release and D glucose utilization or oxidation in the presence of 3-O-methyl-D-glucose was not different from that otherwise observed at increasing concentrations of either D glucose or D-mannoheptulose. It is concluded, therefore, that 3-O-methyl-D glucose adversely affects the metabolism and insulinotropic action of D-glucose by a mechanism largely unrelated to changes in the intracellular concentration of the latter hexose. PMID- 10391132 TI - Effect of palmitate on carbohydrate utilization and Na/K-ATPase activity in aortic vascular smooth muscle from diabetic rats. AB - Several investigators have reported that carbohydrate metabolism is suppressed in blood vessels from diabetic (Db) rats. However, it is not known if metabolites from the reciprocal increase in oxidation of long-chain fatty acids that accompanies insulin-deficiency exacerbates the suppression of this pathway in the Db blood vessels. Such inhibition may have particularly deleterious consequences in vascular smooth muscle since aerobic glycolysis is believed to preferentially fuel the sarcolemmal Na/K ATPase in this tissue. Therefore, this study evaluated the effect of physiological (0.4 mM) and elevated (1.2 mM) concentrations of the long-chain fatty acid palmitate on both carbohydrate utilization and Na/K-ATPase activity in aorta from insulin-deficient Db rat. Thoracic aorta were removed from 10 week Db (streptozotocin 60 mg/Kg , i.v.) or control (C) rats and intima-media aortic preparations were incubated in the absence or presence of palmitate. Glycolysis (microM/g dry wt/h) and glucose oxidation (microM/g dry wt/h) were quantified using 3H-glucose and 14C-glucose, respectively. Na/K-ATPase activity was estimated by the measurement of 86rubidium uptake in the absence and presence of 2 mM ouabain. In the absence of exogenous palmitate, glycolysis (p < 0.05), glucose oxidation (p < 0.01) and the estimated ATP production from exogenous glucose were decreased in aorta from Db rat. However, despite this diminished rate of glycolysis, Na/K ATPase activity was similar in Db and C aorta. Palmitate (0.4 mM) inhibited Na/K ATPase activity and glucose oxidation to a similar extent in both Db and C but had no effect on glycolysis in either group. Elevation of palmitate to 1.2 mM had no additional inhibitory effect on glucose oxidation, Na/K ATPase activity or glycolysis in either the Db or C aorta. The metabolism of exogenous palmitate restored the ATP production in Db to control values. These data demonstrate that, despite the diminished glycolysis and glucose oxidation demonstrated in the Db tissue, Na/K ATPase activity was comparable in the C and Db aorta, in the absence or presence of exogenous long-chain fatty acid. Therefore, the accelerated oxidation of palmitate in diabetic vascular smooth muscle had no additional inhibitory effect on glycolysis or Na/K ATPase activity. These data suggest that Na/K ATPase activity in vascular smooth muscle is not impaired by the altered pattern of substrate utilization that occurs in insulin deficient Db rats. PMID- 10391134 TI - A single cell model of myocardial reperfusion injury: changes in intracellular Na+ and Ca2+ concentrations in guinea pig ventricular myocytes. AB - To investigate the contribution of the changes in intracellular Na+ and Ca2+ concentrations ([Na+]i and [Ca2+]i) to myocardial reperfusion injury, we made an ischemia/reperfusion model in intact guinea pig myocytes. Myocardial ischemia was simulated by the perfusion of metabolic inhibitors (3.3 mM amobarbital and 5 microM carbonyl cyanide m-chlorophenylhydrazone) with pH 6.6 and reperfusion was achieved by the washout of them with pH 7.4. [Na+]i increased from 7.9 +/- 2.0 to 14.0 +/- 3.4 mM (means +/- S.E., p < 0.01 ) during 7.5 min of simulated ischemia (SI) and increased further to 18.8 +/- 3.0 mM at 7.5 min after reperfusion. [Ca2+]i, expressed as the ratio of fluo 3 fluorescence intensity, increased to 133 +/- 8% (p < 0.01) during SI and gradually returned to the control level after reperfusion. Intracellular pH decreased from 7.53 +/- 0.04 to 6.31 +/- 0.04 (p < 0.01) and recovered quickly after reperfusion. Reperfusion with the acidic solution or the continuous perfusion of hexamethylene amiloride (2 microM) prevented the reperfusion-induced increase in [Na+]i. When the duration of SI was prolonged to 15 min, the cell response after reperfusion varied, 16 of 37 cells kept quiescent, 21 cells showed spontaneous Ca2+ waves, and 4 cells out of these 21 cells became hypercontracted. In quiescent cells, both [Na+]i and [Ca2+]i decreased immediately after reperfusion. In cells with Ca2+ waves, [Na+]i transiently increased further at the early phase of reperfusion, while [Ca2+]i declined. In hypercontracted cells, [Na+]i increased as much as in 'Ca2+ wave' cells, but [Ca2+]i increased extensively and both ion concentrations continued to increase. Reperfusion with the Ca2+-free solution prevented both the [Ca2+]i increase and morphological change. In the presence of ryanodine (10 microM), the increase in [Ca2+]i after reperfusion was augmented and some cells became hypercontracted. We concluded that (1) Na+/H+ exchange is active both during SI and reperfusion, resulting in the additional [Na+]i elevation on reperfusion, (2) the [Na+]i level after reperfusion and the following Ca2+ influx via Na+/Ca2+ exchange are crucial for reperfusion cell injury, and (3) the Ca2+ buffering capacity of sarcoplasmic reticulum would also contribute to the Ca2+ regulation and cell injury after reperfusion. PMID- 10391135 TI - Effects of peroxide on contractility of coronary artery rings of different sizes. AB - Reactive oxygen species (ROS, free radicals) produced during cardiac ischemia and reperfusion can damage the contractile functions of arteries. The sarcoplasmic reticulum (SR) Ca2+ pump in coronary artery smooth muscle is very sensitive to ROS. Here we show that contractions of de-endothelialized rings from porcine left coronary artery produced by the hormone Angiotensin II and by the SR Ca2+ pump inhibitors cyclopiazonic acid and thapsigargin correlate negatively with the tissue weight. In contrast, the contractions due to membrane depolarization by high KCl correlate positively. Peroxide also produces a small contraction which correlates negatively with the tissue weight. When artery rings are treated with peroxide and washed, their ability to contract with Angiotensin II, cyclopiazonic acid and thapsigargin decreases. Thus, the SR Ca2+ pump may play a more important role in the contractility of the smaller segments of the coronary artery than in the larger segments. These results are consistent with the hypothesis that ROS which damage the SR Ca2+ pump affect the contractile function of the distal segments more adversely than of the proximal segments. PMID- 10391136 TI - Mitochondrial creatine kinase functional development in post-natal rat skeletal muscle. A combined polarographic/31P NMR study. AB - Mitochondrial creatine kinase (Mi-CK) function in viable mitochondria from developing rat skeletal muscle was assessed both by polarographic measurements of creatine-induced respiration and 31P NMR spectroscopy measurements of phosphocreatine (PCr) synthesis. Creatine-induced respiration was observed in very young rats and increased by 50% to 35 days of age. PCr synthesis was present in 7 day old animals and increased by 300% reaching levels measured in 35 day and adult muscle. Unlike reports showing Mi-CK enzymatic activities but no mitochondrial function in several situations, a concomitant progression of enzymatic activity and mitochondrial function was evidenced during the developmental stages of skeletal muscle Mi-CK in altricious animals. These results correlated with the progressive pattern of muscle differentiation during development of motricity in such animals. The observation that Mi-CK is functional in skeletal muscle mitochondria very early after birth, strongly favors the notion that adaptations in skeletal muscle of Mi-CK knock-out mice occur early. PMID- 10391137 TI - Inhibition of calcium ATPase by phencyclidine in rat brain. AB - Phencyclidine (PCP) is a potent psychotomimetic drug of abuse and has profound effect on the functioning of the central nervous system (CNS). Many of the CNS functions are known to be mediated by calcium (Ca2+). In the present study we have investigated the effects of PCP on Ca2+ ATPase activity in rat brain both in vitro and in vivo. For in vitro studies, synaptic membrane fractions prepared from normal rat brain were incubated with PCP at different concentrations (25-100 microM) before the addition of substrate. For in vivo studies, rats were treated with a single moderate dose of PCP (10 mg/kg, i.p.) and animals were sacrificed at 1,2, 6 and 12 h after treatment. Ca2+ ATPase activity in synaptic membrane fractions was assayed by estimation of inorganic phosphate. PCP inhibited the Ca2+ ATPase in vitro in a concentration dependent manner with significant effect at 50 and 100 microM. A significant time-dependent reduction of the Ca2+ ATPase activity was evident in vivo. As early as 2 h after the treatment of rats with PCP the ATPase activity was significantly reduced. The reduction of Ca2+ ATPase observed even at 12 h after treatment suggesting a prolonged presence of the drug in the brain tissue. Further, kinetic studies in vitro indicated PCP to be a competitive inhibitor of Ca2+ ATPase with respect to the substrate, ATP. The present findings indicate that PCP inhibits synaptic membrane Ca2+ ATPase thus altering cellular Ca2+ homeostasis in CNS which may partially explain the pharmacological effects of the drug and/or its neurotoxicity. PMID- 10391138 TI - Studies on hepatic oxidative stress and antioxidant defence system during chloroquine/poly ICLC treatment of Plasmodium yoelii nigeriensis infected mice. AB - Reactive oxygen species are important mediators of tissue injury during malaria infection. The status of hepatic oxidative stress and antioxidant defence indices were studied during Plasmodium yoelii nigeriensis (P. y. nigeriensis) infection and chloroquine/ polyinosinic-polycytidylic acid stabilized with polylysine and carboxymethylcellulose (poly ICLC) treatment of infected mice. P. y. nigeriensis infection resulted in a significant increase in oxidative stress indices viz., xanthine oxidase and rate of lipid peroxidation (LPO). This was accompanied by a highly significant increase in antioxidant defence indices viz., reduced glutathione (GSH) and glutathione reductase while superoxide dismutase (SOD) and catalase showed a highly significant decrease with respect to normal mice. Chloroquine treatment of infected mice caused a decrease in parasitaemia which was associated with restoration of indices altered during infection towards normalization. Poly ICLC treatment of infected mice caused no change in blood parasitaemia but resulted in a significant increase in GSH, glutathione reductase, SOD and catalase with respect to infected mice. Combination therapy of chloroquine and poly ICLC resulted in clearance of parasitaemia and restoration of all oxidative stress and antioxidant defence indices to normal levels. PMID- 10391139 TI - Differences in the reducing power along the rat GI tract: lower antioxidant capacity of the colon. AB - The ability of the gastrointestinal (GI) tract, as well as other tissues, to cope with reactive oxygen species (ROS) efflux in pathological events is determined partly by epithelial antioxidant levels. These levels are comprised of tissue antioxidant enzymes and low molecular weight antioxidants (LMWA). While glutathione levels and the activity of enzymatic antioxidants along the GI tract have been studied, the contribution of the overall LMWA to the total antioxidant capacity has not yet been determined. In this study the overall antioxidant activity in the mucosa/submucosa and muscularis/serosa of various sections along the small intestine and colon of the rat was evaluated by determining the reducing power, which reflects the total antioxidant activity derived from LMWA, using cyclic voltammetry. The activity of the antioxidant enzymes superoxide dismutase (SOD) and catalase was also measured. The reducing power (total antioxidant activity) was higher in the mucosa/submucosa of the small intestine as compared to the mucosa/submucosa of the colon. Similarly, catalase and SOD activity in the mucosa/submucosa of the small intestine was significantly higher than in the mucosa/submucosa of the colon. Differences were also observed in the reducing power and SOD activity in the muscularis/serosa of the rat small intestine as compared to the colon. The low antioxidant capacity in the colon may facilitate reactive oxygen species (ROS)-mediated injury and lead to inflammatory diseases such as ulcerative colitis, specifically in the colon. PMID- 10391140 TI - The relation between the action potential duration, the increase in resting tension, and ATP content during metabolic inhibition in guinea pig ventricular muscles. AB - To investigate whether the action potential duration (APD) or resting tension was dependent on global ATP content, and whether they were preferentially dependent on glycolytic ATP, APD and resting tension were measured under various metabolic inhibition with corresponding measurement of ATP content in guinea pig ventricular muscles. Oxidative phosphorylation was inhibited by either hypoxic perfusion, the perfusion of sodium cyanide, or 2,4-dinitrophenol. Glycolysis was blocked by the perfusion of iodoacetic acid, and hypoxia with variable glycolytic activities was achieved by hypoxic perfusion in the presence of glucose (5, 10, and 50 mM). APD began to decrease when ATP content decreased to less than 3 mM/kg w.w. from the control level of 4.35 mM/kg w.w. APD shortened significantly and resting tension increased steeply, when ATP content decreased below 1 mM/kg w.w. The dependence of APD and the increase in resting tension on ATP content was not affected by the mode of metabolic block, that is, the inhibition of glycolysis and/or oxidative phosphorylation. Though other factors can affect APD and resting tension, we found no evidence of functional ATP compartmentation, with respect to APD and the increase in resting tension during metabolic inhibition. PMID- 10391141 TI - Identification of the histamine H1 receptor gene as a differentially repressed target of the human TR2 orphan receptor. AB - We have identified a DNA response element (TR2RE-HR) in the 3' flanking region of the human histamine H1 receptor gene as a target for the TR2 orphan receptor, a member of the steroid/thyroid hormone receptor superfamily. The application of both tetracycline inducible and improved differential display systems has allowed us to isolate a cDNA fragment differentially regulated by the expression of the TR2 orphan receptor. Northern blot and sequencing analysis demonstrated that the expression of the human histamine H1 receptor gene was differentially repressed by the TR2 orphan receptor. Electrophoretic mobility shift assay further revealed a specific binding (dissociation constant = 26.2 nM) between the TR2 orphan receptor and the wildtype TR2RE-HR, but not the mutant TR2RE-HR. In addition, reporter gene expression assay indicated that the TR2 orphan receptor may suppress the expression of luciferase activities in a dose-dependent manner via the TR2RE-HR in HeLa cells. Our results demonstrate that the histamine H1 receptor gene could represent one of the target genes directly regulated by the human TR2 orphan receptor. PMID- 10391142 TI - Dephosphorylation increases insulin-stimulated receptor kinase activity in skeletal muscle of obese Zucker rats. AB - Serine/threonine phosphorylation of insulin receptor has been implicated in the development of insulin resistance. To investigate whether dephosphorylation of serine/threonine residues of the insulin receptor may restore the decreased insulin-stimulated receptor tyrosine kinase activity in skeletal muscle of obese Zucker rats, insulin receptor tyrosine kinase activity was measured before and after alkaline phosphatase treatment. Compared to lean controls, insulin stimulated glucose transport was depressed by 61% (p < 0.05) in obese Zucker rats. The insulin receptor and insulin receptor substrate-1 contents were decreased by 14% (p < 0.05) and 16% (p < 0.05), respectively, in skeletal muscle of obese Zucker rats. In vivo insulin-induced tyrosine phosphorylation of insulin receptor and insulin receptor substrate-1 was depressed by 82% (p < 0.05) and 86% (p < 0.05), respectively. In the meantime, in vitro insulin-stimulated receptor tyrosine kinase activity in obese rats was decreased by 39% (p < 0.05). Dephosphorylation of the insulin receptor by prior alkaline phosphatase treatment increased insulin-stimulated receptor tyrosine kinase activity in both lean and obese Zucker rats, but the increase was three times greater in obese Zucker rats (p < 0.05). These findings suggest that excessive serine/threonine phosphorylation of the insulin receptor in obese Zucker rats may be a cause for insulin resistance in skeletal muscle. PMID- 10391143 TI - Alpha-tocopherol inhibits oxidative stress induced by cholestanetriol and 25 hydroxycholesterol in porcine ovarian granulosa cells. AB - The cytotoxicity of oxysterols including 7-ketocholesterol, alpha-epoxide, cholestanetriol and 25-hydroxycholesterol and the possible protecting effect of alpha-tocopherol on cholestanetriol and 25-hydroxycholesterol-induced cytotoxicity were investigated in primary cultures of porcine ovarian granulosa cells. Cell viability as determined by % trypan blue staining and mitochondrial function as determined using 3-[4,5-dimethylthiazol-2-yl]-2,5- diphenyltetrazolium bromide (MTT) reduction were decreased significantly after 24 h exposure to 2.5 microM alpha-epoxide, cholestanetriol and 25 hydroxycholesterol. 7-Ketocholesterol (2.5 microM) did not affect cell viability or mitochondrial function under the same culture conditions. The specific activities of catalase and superoxide dismutase, two antioxidant defense enzymes were increased significantly (p < 0.01) following 24 h exposure to 2.5 microM concentrations of cholestanetriol while only superoxide dismutase was increased in 25-hydroxycholesterol-treated cells (p < 0.001). Specific activity of glutathione peroxidase was unchanged relative to control cells. Levels of thiobarbituric acid reactive substances remained unchanged after exposure to 7 ketocholesterol, alpha-epoxide, cholestanetriol, 25-hydroxycholesterol and cholesterol. Administration of 1 microM alpha-tocopherol to the culture medium significantly improved cell viability and restored both superoxide dismutase and catalase activities to control levels in cholestanetriol -treated cells and only superoxide dismutase in 25-hydroxycholesterol-treated cells. These studies suggest that the cytotoxic nature of physiologically relevant concentrations of cholestanetriol and 25-hydroxycholesterol in granulosa cells is in part due to oxidative stress, but it may be reduced in the presence of alpha-tocopherol. PMID- 10391144 TI - Increased peroxisomal fatty acid beta-oxidation and enhanced expression of peroxisome proliferator-activated receptor-alpha in diabetic rat liver. AB - To determine whether the increased fatty acid beta-oxidation in the peroxisomes of diabetic rat liver is mediated by a common peroxisome proliferation mechanism, we measured the activation of long-chain (LC) and very long chain (VLC) fatty acids catalyzed by palmitoyl CoA ligase (PAL) and lignoceryl CoA ligase and oxidation of LC (palmitic acid) and VLC (lignoceric acid) fatty acids by isotopic methods. Immunoblot analysis of acyl-CoA oxidase (ACO), and Northern blot analysis of peroxisome proliferator-activated receptor (PPAR-alpha), ACO, and PAL were also performed. The PAL activity increased in peroxisomes and mitochondria from the liver of diabetic rats by 2.6-fold and 2.1 -fold, respectively. The lignoceroyl-CoA ligase activity increased by 2.6-fold in diabetic peroxisomes. Palmitic acid oxidation increased in the diabetic peroxisomes and mitochondria by 2.5-fold and 2.7-fold, respectively, while lignoceric acid oxidation increased by 2.0-fold in the peroxisomes. Immunoreactive ACO protein increased by 2-fold in the diabetic group. The mRNA levels for PPAR-alpha, ACO and PAL increased 2.9-, 2.8- and 1.6-fold, respectively, in the diabetic group. These results suggest that the increased supply of fatty acids to liver in diabetic state stimulates the expression of PPAR-alpha and its target genes responsible for the metabolism of fatty acids. PMID- 10391145 TI - Involvement of signal transduction pathways in Salmonella typhimurium porin activated gut macrophages. AB - Many membrane proteins are implicated in the regulation of cell functions by triggering specific signaling pathways. Porins are known potential modulators of cell proliferation and differentiation. We explored the possible involvement of this protein in signal transduction pathways in mouse gut macrophages. In the present work we have shown that porins can trigger signal transduction in mouse macrophages infected with S. typhimurium. Activation of macrophages by porins results in an increase in inositol trisphosphate and intracellular Ca2+ mobilization. There is a translocation of protein kinase C to the membrane which is accompanied by nitric oxide release within the macrophages. This effect is the outcome of the expression of nitric oxide synthase, which is dependent on Protein kinase C. Further, we observed that there is an increased binding of the porins on macrophages infected with S. typhimurium which results in activation of macrophages and triggering of specific signaling pathways. These results indicate that porins induce the production of nitric oxide via a protein kinase C dependent pathway. Nitric oxide plays a fundamental role in macrophage effector function where it has both communication and defensive function. PMID- 10391146 TI - Biochemical consequences of electrical pacing in ischemic-reperfused isolated rat hearts. AB - It is still unclear if performance recovery in postischemic hearts is related to their tissue level of high-energy phosphates before reflow. To test the existence of this link, we monitored performance, metabolism and histological damage in isolated, crystalloid-perfused rat hearts during 20 min of low-flow ischemia (90% coronary flow reduction) and reflow. To prevent interference from different ischemia times and perfusing media compositions, the ischemic ATP level was varied by changing energy demand (electrical pacing at 330 min(-1)). Under full coronary flow conditions, work output, as well as ATP and phosphocreatine contents were the same in control, spontaneously contracting (n = 23) and paced (n = 21) hearts. During low-flow ischemia, the higher work output (p < 0.0001) in paced hearts decreased their tissue content of ATP, phosphocreatine and total adenylates and purines (p < 0.05), as opposed to maintained values in control hearts. During reflow, the recovery of mechanical performance and O2 uptake was 94 +/- 5% and 110 +/- 9% (p = NS vs. baseline) in controls, vs. 71 +/- 5% and 74 +/- 6% in paced hearts (p < 0.004 vs. baseline). The levels of ATP and total adenylates and purines remained constant in control, but were markedly depressed (p < 0.05 vs. baseline) in paced hearts. Phosphocreatine+creatine was the same in both groups. These data, together with the observed lack of creatine kinase leakage and of structural damage, indicate that myocardial recovery during reflow reflects the tissue level of ATP, phosphocreatine and total adenylates and purines during ischemia, regardless of physical cell damage. PMID- 10391147 TI - The Ca2+ threshold for the mitochondrial permeability transition and the content of proteins related to Bcl-2 in rat liver and Zajdela hepatoma mitochondria. AB - Zajdela hepatoma mitochondria were able to accumulate two to five times more Ca2+ than rat liver mitochondria before the permeability transition was induced. Pulses of Ca2+ were given in series to determine the Ca2+ threshold by recording changes in [Ca2+] and membrane potential, the permeability transition causing the release of accumulated Ca2+ and collapse of the membrane potential. Hepatoma mitochondria had lower Ca2+ efflux rates, higher net Ca2+ uptake rates and lower phosphorylation rates than liver mitochondria. Since the differences in regard to induction of the permeability transition might be due to higher expression of the Bcl-2 protein in hepatoma cells than in hepatocytes, the transcription of Bcl-2 and the proteins reacting with a Bcl-2 polyclonal antiserum were estimated by Northern and Western blotting, respectively. Hepatoma cells had two Bcl-2 specific mRNA bands of 7 and 2.4 kb, and substantial amounts of the Bcl-2 protein, whereas in liver cells and mitochondria these were not detected. Both cell lines had a reactive band at 19-20 kDa, and hepatocytes a small band at 31 32 kDa. Bcl-2 antibodies stimulated the permeability transition potently in hepatoma mitochondria. PMID- 10391148 TI - An in vitro study on the role of metal catalyzed oxidation in glycation and crosslinking of collagen. AB - The present investigation was carried out to understand the effect of metal catalyzed oxidation on glycation and crosslinking of collagen. Tail tendons obtained from rats weighing 200-225 g were incubated with glucose (250 mM) and increasing concentrations of copper ions (5, 25, 50 and 100 microM) under physiological conditions of temperature and pH. Early glycation, crosslinking and late glycation (fluorescence) of collagen samples were analyzed periodically. Early glycation was estimated by phenol sulfuric acid method, and the crosslinking was assessed by pepsin and cyanogen bromide digestion. A concentration-dependent effect of metal ions on the rate of glycation and crosslinking of collagen was observed. Tendon collagen incubated with glucose and 100 microM copper ions showed 80% reduction in pepsin digestion within seven days, indicating extensive crosslinking, whereas collagen incubated with glucose alone for the same period showed only 7% reduction. The presence of metal ions in the incubation medium accelerated the development of Maillard reaction fluorescence on collagen, and the increase was dependent on the concentration of metal ions used. The metal chelator diethylene triamine penta-acetate significantly prevented the increase in collagen crosslinking by glucose and copper ions. Free radical scavengers benzoate and mannitol effectively prevented the increased crosslinking and browning of collagen by glucose. The results indicate that the metal catalyzed oxidation reactions play a major role in the crosslinking of collagen by glucose. It is also suggested that the prevention of increased oxidative stress in diabetes may prevent the accelerated advanced glycation and crosslinking of collagen. PMID- 10391149 TI - Possible involvement of Ca2+/calmodulin-dependent protein kinase in the regulation of phospholipid biosynthesis in Microsporum gypseum. AB - The mechanism of action of Ca2+/calmodulin on phospholipid synthesis in Microsporum gypseum has been studied. These second messengers were observed to mediate their function through phosphorylation mechanism as altered protein kinase activity was seen in calcium/trifluoperazine (calmodulin antagonist) grown cells. The activity of protein kinase was dependent on calcium (200 microm) and calmodulin (1 microm). In vitro studies of phosphorylation and dephosphorylation in relation to phospholipid synthesis in Microsporum gypseum have been carried out. Addition of KN-62 (a specific inhibitor of Ca2+/calmodulin-dependent protein kinases) and polyclonal antibodies raised against purified Ca2+/calmodulin-kinase (CaMPK) of M. gypseum in the cell extract, leads to the inhibition in the incorporation of labelled acetate into total phospholipids in this fungus. These results suggest a possible involvement of Ca2+/calmodulin via Ca2+/calmodulin dependent phosphorylation in phospholipid synthesis in M. gypseum. PMID- 10391150 TI - Picroliv -- a natural product protects cells and regulates the gene expression during hypoxia/reoxygenation. AB - Cellular adaptation to hypoxia involves regulation of specific genes such as vascular endothelial growth factor (VEGF), erythropoietin (EPO) and hypoxia inducible factor (HIF)-1 . In this study, we have evaluated the protective effect of picroliv (a purified iridoid glycoside fraction from roots of Picrorhiza kurrooa with hepatoprotective, anti-inflammatory and antioxidant properties) against hypoxic injury by examining lactate dehydrogenase (LDH) release in Hep 3B and Glioma cells. The expression of hypoxia regulated genes, VEGF and HIF-1 was studied in human umbilical vein endothelial cells (HUVEC), Hep 3B and Glioma cells. Picroliv reduced the cellular damage caused by hypoxia as revealed by a significant reduction in LDH release compared to untreated control. The expression of VEGF and HIF-1 subunits (HIF-1alpha and HIF-1beta) was enhanced by treatment with picroliv during normoxia and hypoxia in HUVEC and Hep 3B cells and on reoxygenation the expression of these genes was significantly reduced as revealed by mRNA analysis using RT-PCR. Simultaneous treatment with picroliv during hypoxia inhibited VEGF and HIF-1 expression in Glioma cells whereas the expression was not reduced by picroliv treatment during reoxygenation as evidenced by both RT-PCR and Northern hybridization. VEGF expression as revealed by immunofluorescence studies correlates well with the regulations observed in the mRNA expression. We have also examined the kinase activity of tyrosine phosphorylated proteins and protein kinase C (PKC) in Glioma cells treated with picroliv during hypoxia/reoxygenation. A selective inhibition of protein tyrosine kinase activity leading to tyrosine dephosphorylation of several proteins including 80 kd protein, and a reduction in PKC was seen in cells treated with picroliv and hypoxia. These findings suggest that picroliv may act as a protective agent against hypoxia/reoxygenation induced injuries, and the underlying mechanism may involve a novel signal transduction pathway. PMID- 10391151 TI - Relation of NADH/NAD to contraction in vascular smooth muscle. AB - The relationship of NADH/NAD to O2 consumption with respect to the different phases of contraction in vascular smooth muscle in response to a maximal depolarizing concentration of KCl was investigated. The NADH bound to cellular proteins could be distinguished from free NADH in whole tissue homogenates. Evidence suggested that the NADH was bound to pyruvate dehydrogenase and perhaps to other dehydrogenases since binding paralleled the changes in the activity of pyruvate dehydrogenase with contraction. The measured changes in NADH were attributed to that within the mitochondrial compartment since the contribution of reducing equivalents within the cytoplasmic compartment was negligible. During the phase of contraction in which force was initially being generated and at which O2 consumption was the highest, there was a net increase in NADH/NAD. After stable isometric force was maintained, at which time O2 consumption had returned to slightly above the basal pre-contraction level, there was a net decrease in NADH/NAD. Previous evidence indicates the phosphorylation potential (ATP/ADP) may decrease during this phase of contraction. It is concluded that contraction of vascular smooth muscle is accompanied by a changing pool of reducing equivalents. Factors which govern O2 consumption may change during the different phases of muscle contraction. PMID- 10391152 TI - Effects of amiodarone on cardiac function and mitochondrial oxidative phosphorylation during ischemia and reperfusion. AB - This study was carried out in order to determine if the efficiency of amiodarone, a class III antiarrhythmic agent, is associated with changes in mitochondrial oxidative phosphorylation. A population of 30 rats were treated with amiodarone (100 mg/kg/day) for 5 days. A second population receiving only vehicle was used as control. The hearts were perfused according to the working mode. After 15 min of normoxic perfusion, the left main coronary artery was ligated and the ligation was maintained for 20 min. The ligation was removed and reperfusion continued for a further 30 min. The electrocardiogram was monitored continuously. At the end of perfusion, the ischemic and non ischemic areas were visually separated and mitochondria were harvested from each area. Their oxidative and energy metabolism were assessed with palmitoylcarnitine as substrate in 2 respiration media differing in their free calcium concentration (0 or 0.34 microm). In normoxic conditions, amiodarone treatment increased the cardiac metabolic efficiency (mechanical work to oxygen consumption ratio). The local ischemia decreased the aortic and coronary flows without modifying the cardiac metabolic efficiency. Amiodarone treatment maintained the aortic flow at a significantly higher value; the duration of severe arrhythmias was significantly decreased by the drug. The reperfusion of the ischemic area allowed the partial recovery of fluid dynamics. The coronary flow was restored to 89% of the pre ischemic value. Conversely, the aortic flow never exceeded that measured at the end of ischemia, partly due to the important development of severe arrhythmias. The recovery of aortic flow and metabolic efficiency during reperfusion was improved by amiodarone treatment; ventricular tachycardia and fibrillation duration were reduced. In the mitochondria issued from the normoxic area, the energy metabolism was not altered by the amiodarone treatment, but the presence of calcium in the respiration medium modified the oxidative phosphorylation. The divalent cation slightly decreased the state III respiration rate and increased noticeably the state IV respiration rate. This was associated with an important mitochondrial AMP production and maintenance of ADP in the respiration medium. This energy wasting was reported to decrease the mitochondrial metabolic efficiency. After an ischemia-reperfusion sequence, mitochondrial oxidation phosphorylation was reduced and amiodarone treatment amplified this decrease. This was presumably due to an increased mitochondrial calcium accumulation. Thus, the beneficial properties of amiodarone during reperfusion are supposed to be due to a protection against the deleterious effect of excess matrix calcium on mitochondrial energy metabolism. PMID- 10391154 TI - Reduction "by half". The need for standardised surgical technique in studies of radiotherapy for rectal cancer. PMID- 10391153 TI - Calcium regulation in the human myocardium affected by dilated cardiomyopathy: a structural basis for impaired Ca2+-sensitivity. AB - Calcium regulation in the human heart is impaired during idiopathic dilated cardiomyopathy (IDC). Here, we analyze the structural basis for impairment in the regulatory mechanism. Regulation of contractility was monitored by MgATPase and Ca2+-binding assays as a function of calcium. Myofibrillar proteolysis and expression of troponin T isoforms were established by gel electrophoresis and by Western blots. Myofibrillar ATPase assays in low salt however, revealed a drastic lowering of calcium sensitivity in IDC myofibrils as indicated by reductions in both activation by high calcium and in EGTA-mediated inhibition of MgATPase. Structural changes in myofilament proteins were found in most IDC hearts, specifically proteolysis of myosin light chain 2 (LC2), troponin T and I (TnT and TnI), and sometimes a large isoform shift in TnT. IDC did not induce mutations in LC2 and troponin C (TnC), as established by cDNA sequence data from IDC cases, thus, calcium binding to IDC myofibrils was unaffected. Reassociation of IDC myofibrils with native LC2 raised MgATPase activation at high Ca2+ to control levels, while repletion with intact, canine TnI/TnT restored inhibition at low Ca2+. A model, identifying possible steps in the steric blocking mechanism of regulation, is proposed to explain IDC-induced changes in Ca2+-regulation. Moreover, shifts in TnT isoforms may imply either a genetic or a compensatory factor in the development and pathogenesis of some forms of IDC. PMID- 10391155 TI - Total mesorectal excision (TME) with or without preoperative radiotherapy in the treatment of primary rectal cancer. Prospective randomised trial with standard operative and histopathological techniques. Dutch ColoRectal Cancer Group. AB - OBJECTIVE: To document local recurrence in primary rectal cancer when standardised techniques of surgery, radiotherapy, and pathology are used, and to investigate whether the local recurrence rate after total mesorectal excision permits the omission of adjuvant short term preoperative radiotherapy. DESIGN: Prospective randomised study. SETTING: Dutch (n = 80), English (n = 1), German (n = 1), Swedish (n = 9), and Swiss (n = 1) hospitals. SUBJECTS: The first 500 randomised Dutch patients with primary rectal cancer. MAIN OUTCOME MEASURES: Local recurrence, survival, operation-related factors, specific pathological tumour characteristics, short and long term morbidity, and quality of life. RESULTS: Between January 1996 and April 1998, 871 Dutch and 94 other patients were randomised. Our feasibility analysis shows that cooperation between and within the participating disciplines goes well. With regard to the surgical part, this can be confirmed by the large number of operations attended by consultant surgeons (58%). The number of abdominoperineal resections appeared to be low (30%), as did the percentage of lateral margins involved (13%). The rate of adverse effects of radiotherapy was acceptable. Apart from a larger operative blood loss and a higher infective complication rate in the irradiated group, no significant differences were found with regard to morbidity and mortality between the randomised groups. CONCLUSIONS: The accrual of our trial is going well and it is feasible; short term preoperative radiotherapy is safe even in combination with TME. PMID- 10391156 TI - Recording and classification of complications in a surgical practice. AB - OBJECTIVE: To document the incidence and outcome of complications in the department of surgery. DESIGN: Retrospective study. SETTING: District hospital, The Netherlands. SUBJECTS: 7455 patients operated on between 1 January 1993 and 31 December 1995. MAIN OUTCOME MEASURES: Documentation and outcome of complications (defined as "every unwanted development in the illness of the patient or in the treatment of the patient's illness that occurs in the clinic"). RESULTS: 1078 complications were recorded after 8130 operations (13%), 337 (33%) of which had no long term effects. 175/1078 (16%) required reoperation, and in 134 of these (77%) an error in management or surgical technique was responsible for the complication. 6 patients were irreversibly harmed and of the 141 patients who died, 11 had evidence of some sort of error. CONCLUSIONS: Audit of complications is necessary to improve practice in a surgical department, and weekly morbidity and mortality meetings are a good opportunity for learning about them. PMID- 10391157 TI - Improved trauma care after reorganisation: a retrospective analysis. AB - OBJECTIVE: To shorten the time to make a diagnosis and to begin definitive treatment of severely injured patients, thereby improving their medical care. DESIGN: Retrospective analysis. SETTING: Teaching hospital, Sweden. SUBJECTS: 61 patients who had sustained high-energy injuries, including head injury which required surgical intervention, and fracture of the femoral shaft before (1987 1988 n = 23) and after (1991-1993 n = 38) the reorganisation. INTERVENTION: Trauma care was reorganised during the year 1989-1990 and the concept of early multidisiplinary treatment with the general surgeon as trauma-leader was adopted. MAIN OUTCOME MEASURES: The time required to make a diagnosis and begin definitive treatment as well as the assessment of medical care taking account of the patient's general condition and other injuries. RESULT: The immediate medical care was classified as delayed or inappropriate in 9 of 23 patients before, and in 2 of 38 patients after, the reorganisation (p = 0.001). The time needed to make a diagnosis was less than 4 hours in all cases. The time needed to start definitive treatment of head injuries was less than four hours in 9 of 12 patients before, and in 18 of 21 patients after the reorganisation. The internal fixation of femoral fractures was started within four hours in 2 of 11 femoral fractures before, compared with 12 of 17, after the reorganisation. CONCLUSION: The time to beginning definitive treatment of severe injuries was shorter after the reorganisation, as a result of early participation of members of the trauma team. PMID- 10391158 TI - Long term experience after subtotal adrenalectomy for multiple endocrine neoplasia type IIa. AB - OBJECTIVE: To evaluate the long-term results after subtotal adrenalectomy in patients with multiple endocrine neoplasia type IIa (MEN IIa). DESIGN: Retrospective study. SETTING: University Hospital, Sweden. SUBJECTS: Five patients who underwent partial adrenalectomy between 1985 and 1989. INTERVENTIONS: Subtotal adrenalectomy with a rim of cortical tissue left in situ. MAIN OUTCOME MEASURES: Follow up by interview, measurement of cortisol and catecholamine excretion in urine, and cortisol concentration in serum in response to stimulation with ACTH. RESULTS: Three patients took no corticosteroids regularly, but during upper respiratory tract infections, or periods of severe stress they took 25 mg cortisone acetate daily. This is confirmed by their normal values of 24 hour urinary cortisol excretion and subnormal responses to an ACTH stimulation test. The fourth and fifth patients had low concentrations of endogenous corticosteroids postoperatively, which is being replaced with 25 mg cortisone acetate daily. Postoperatively all five patients had low urinary adrenaline excretion. CONCLUSION: Subtotal adrenalectomy in patients with MEN IIa resulted in basal endogenous corticosteroids within the reference range in three of five patients. There was no evidence of reduced adrenocortical function with time, nor were there any signs of recurrence of the pheochromocytoma. PMID- 10391159 TI - Association between the number of vascular operation on the lower limbs and long term survival. AB - OBJECTIVE: To investigate to what extent the need for more than one vascular operation for chronic lower limb ischaemia was associated with relative long term survival. DESIGN: Retrospective observational study. SETTING: University hospital, Norway. SUBJECTS: 1574 patients (29% women) operated on for chronic lower limb arterial insufficiency. Of these 447 needed at least one further operation for progressive limb ischaemia. MAIN OUTCOME MEASURES: Long term survival estimated by the Kaplan-Meier method. The expected survival was calculated from mortality tables issued by the Norwegian Central Bureau of Statistics. RESULTS: The 10-year survival rate was 46% for the patients operated on once and 24% for the patients who had two or more operations. The expected survival rates were 57% and 52%, respectively. Both categories of patients had significantly shorter long term survival than a demographically-matched population. The long term survival of patients operated on twice or more was significantly less than that of those who needed only one operation. CONCLUSION: There is an association between the need for more than one vascular operation and long-term survival. Atherosclerotic disease among these patients seems to be more aggressive. PMID- 10391160 TI - Is organ ischaemia a determinant of the outcome of operations for suprarenal aortic aneurysms? AB - OBJECTIVE: To find out if morbidity and mortality after thoracoabdominal approaches for suprarenal aortic aneurysms are related to the duration of organ ischaemia. DESIGN: Retrospective study. SETTING: University hospital, The Netherlands. SUBJECTS: 72 operations for suprarenal aortic aneurysms. MAIN OUTCOME MEASURES: Duration of organ ischaemia, morbidity and mortality. RESULTS: There were 72 patients with 3 group A (Crawford type III), 10 group B (Crawford type IV), 37 group C (supracoeliac), and 22 group D (suprarenal) aneurysms. Median duration of ischaemia was 57 minutes for both the spinal cord and the mesenteric arteries, and 59 and 63 minutes for the right and left renal arteries, respectively. There were 52 major complications in 33 patients. Mesenteric ischaemia of longer than 60 minutes was associated with a significant higher complication rate (21/32, 66% compared with 13/40, 33%, p = 0.01). Spinal cord ischaemia of longer than 60 minutes was not associated with a significantly increased incidence of paraplegia (2/40 compared with 6/32, p = 0.13). CONCLUSIONS: We conclude that with surgery for suprarenal aneurysms a significant higher complication rate is noted with increased duration of mesenteric ischaemia. PMID- 10391161 TI - Morbidity of major hepatic resections: a 100-case prospective study. AB - OBJECTIVE: To assess the morbidity and its main risk factors after major hepatic resection. DESIGN: Retrospective study of prospectively collected data. SETTING: University hospital, France. SUBJECTS: 100 consecutive patients who underwent major hepatic resections, 1989-95. INTERVENTIONS: Major hepatic resection, defined as resection involving 3 or more segments according to Couinaud's classification, in all cases. MAIN OUTCOME MEASURES: All complications that affected outcome or prolonged hospital stay. Risk factors identified by univariate and multivariate analysis. RESULTS: 45 patients developed at least 1 complication and 7 died. The most common complications were: pleural effusion (n = 21), hepatic failure (n = 12), and ascites (n = 9). Univariate analysis showed that the following variables were significantly related to the morbidity: age >55 years, American Society of Anesthesiologists (ASA) grade II or more, bilirubin >80 micromol/L, alkaline phosphatase activity more than double the reference range, malignant tumours, abnormal liver parenchyma, simultaneous surgical procedures, operative time >4 hours, and perioperative blood transfusion > or =600 ml. The extent of resection did not correlate with postoperative complications. Multivariate analysis showed that volume of blood transfusion > or =600 ml and simultaneous surgical procedures were the most important independent risk factors for complicated outcome. CONCLUSIONS: The morbidity associated with major hepatic resections remains high, and the main determinants of outcome are intraoperative surgeon-related factors. PMID- 10391162 TI - Liver transplantation in Japanese and Australian/New Zealand children with biliary atresia: a 10-year comparative study. AB - OBJECTIVE: To compare Japanese with Australian/New Zealand (ANZ) children with biliary atresia who were treated by liver transplantation, and evaluate the indications for and timing of transplantation. DESIGN: Retrospective study. SETTING: Queensland Liver Transplant Service (QLTS), Australia. SUBJECTS: 43 Japanese and 30 ANZ children with biliary atresia who required transplantation between 1985 and 1992. INTERVENTIONS: The 43 Japanese children had 52 transplants, and the 30 ANZ children had 33. MAIN OUTCOME MEASURES: Morbidity, mortality, and long term survival. RESULTS: The Japanese children had significantly lower serum albumin concentrations than the ANZ children preoperatively (mean (SD) 32 (7) g/L compared with 37 (5), p<0.05). The actuarial survival at 7 years of the ANZ children was significantly higher than that of the Japanese children (79% compared with 49%, p<0.05). There were 24 deaths (17 Japanese, 40%, and 7 ANZ, 23%); 2 of the ANZ and 7 of the Japanese children died more than a year after transplantation. All 26 children who were well-nourished at the time of transplantation defined as a Z-score (weight or height minus mean weight or height for age, sex, and race, divided by the SD) of -1 or more were alive at 1 month compared with 11 of the 47 poorly-nourished children (Z-score < 1). Survival among the Japanese declined after 1 year, and there was no association with Z-scores. Overall, Z-scores for weight improved significantly after transplantation, whereas those for height improved a little, but not significantly so. Japanese children were significantly shorter than ANZ children, and their Z-scores for height did not improve after transplantation. CONCLUSION: liver transplantation should be done as soon as possible for children with biliary atresia to maximise survival and growth. PMID- 10391163 TI - Repeated boluses of local anaesthetic for pain relief after inguinal hernia repair. AB - OBJECTIVE: To compare the effect of repeated boluses of local anaesthetics with an oral analgesic for pain management after tension-free inguinal hernia repair. DESIGN: Prospective randomised study. SETTING: University hospital, Germany. SUBJECTS: 104 patients undergoing elective hernia repair. INTERVENTIONS: 52 patients were given boluses of 0.5% bupivacaine 10 ml through a subcutaneous catheter and 52 dipyrone 500 mg orally 6, 12 and 24 hours after operation. MAIN OUTCOME MEASURES: Postoperative pain measured on a visual analogue scale, complications, systemic side effects, supplementary analgesics, costs, and time taken to give the analgesics. RESULTS: There were no significant differences between the groups in absolute pain scores, course of pain, and the effects of analgesics. Thirteen patients [5 in the bupivacaine group and 8 in the dipyrone group] required additional dipyrone [mean (range) 2000 mg (500-5000) and 2500 mg (500-4000) respectively]. There were no systemic side effects of either treatment. There were 5 wound haematomas in the bupivacaine group (10%), and 2 wound haematomas and 1 superficial wound infection in the dipyrone group (6%). Mean costs of material and time taken to give the analgesics were pound sterling 104.70 and 47 minutes in the bupivacaine group compared with pound sterling 3.40 and 6 minutes in the dipyrone group. The median (range) hospital stay was 2 (1-3) days in both groups. CONCLUSIONS: Repeated boluses of local anaesthetic did not result in better or cheaper pain control than oral analgesics after tension-free inguinal hernia repair. PMID- 10391164 TI - Early discharge after appendicectomy in children. AB - OBJECTIVE: To confirm that "day-care" appendicectomy is safe in children. DESIGN: Prospective study. SETTINGS: Paediatric hospitals, Brazil. SUBJECTS: 144 children who required removal of the appendix. INTERVENTIONS: Standard muscle splitting appendicectomy. MAIN OUTCOME MEASURES: Hospital stay, use of antibiotics, and complications. RESULTS: 124 patients (86%) were discharged within 24 hours of operation. In all patients the appendicitis was confirmed and grouped according to histopathological findings (72 inflamed, 26 gangrenous and 26 perforated). 86 patients (60%) were given metronidazole one hour before operation as prophylaxis against postoperative abscess formation. 78 (54%) were given no antibiotics in the postoperative period. 2 patients were readmitted for drainage of intraperitoneal abscesses and 10 had subcutaneous abscesses drained as outpatients during the early postoperative period. The overall infective complication rate was 8% (n = 12), which is comparable with large series of appendicectomy in children. CONCLUSIONS: "Day-care" appendicectomy is safe and feasible in children, as it avoids a long hospital stay and increased costs with no additional risk. PMID- 10391165 TI - Treatment of pilonidal sinus by primary closure with a transposed rhomboid flap compared with deep suturing: a prospective randomised clinical trial. AB - OBJECTIVE: To assess two techniques of primary closure after excision of pilonidal sinus. DESIGN: Prospective randomised study. SETTING: University department of surgery, United Arab Emirates. SUBJECTS: 46 patients with chronic pilonidal sinus disease, 24 treated by rhomboid flap transposition, and 22 by deep suturing technique. MAIN OUTCOME: Early mobility and recurrence. RESULTS: All patients in the rhomboid flap transposition group healed their wounds primarily compared with 17 in the primary deep suturing group (77%). (P = 0.02). Five patients wounds broke down as a result of haematoma and infection (23%). The mean hospital stay for the rhomboid flap technique was 6 days compared with 9 days after deep suturing, and the mean follow up for both groups was 18 months, the rhomboid flap group returned to work a mean of nine days earlier than the deep suturing group (23 days). No recurrence has been identified yet in the rhomboid flap group, while 2 recurrences have developed in the deep suturing group (9%). CONCLUSION: Primary closure after excision of pilonidal sinus with a transposed rhomboid flap is successful in the management of pilonidal sinus and is superior to primary closure by deep suturing. PMID- 10391166 TI - Perforating appendicitis: is it a separate disease? Acute Abdominal Pain Study Group. AB - OBJECTIVE: To find out whether perforated and unperforated appendicitis are separate diseases and can be distinguished clinically. DESIGN: Prospective multicentre study. SETTING: 11 departments of surgery in Germany and Austria. SUBJECTS: 519 patients over 6 years old who had histologically confirmed acute appendicitis between October 1994 and March 1996. MAIN OUTCOME MEASURES: Differences in history, clinical findings, lab results, clinical course and outcome. RESULTS: 92 of the 519 patients (18%) had perforated appendicitis. The following variables were shown by univariate analysis to be significantly more common in the group with perforated appendicitis: rigidity, reduced abdominal wall movement, abdominal distension, reduced bowel sounds (all p<0.001), pale skin (p<0.005), generalised abdominal tenderness, severe abdominal tenderness (both p<0.01), WCC > or =10(9)/L (p<0.05). By multivariate analysis the following variables were significantly more common in the group with perforated appendicitis: age over 50 years (p<0.0001); change in bowel habit and rigidity of the abdominal wall (both p = 0.001); generalised tenderness (p<0.01); male sex (p<0.01); and distended abdomen (p<0.05). Rectal examination failed to make the distinction. CONCLUSIONS: Perforated and unperforated appendicitis behave clinically like two different diseases. They can in most cases reliably be distinguished using clinical criteria alone. Although greater diagnostic accuracy may result in a higher rate of perforation, close observation and timely intervention will only marginally affect the outcome. PMID- 10391167 TI - "Can perforating appendicitis be considered a separate disease entity"? PMID- 10391168 TI - Potential mechanisms responsible for zymosan-associated endothelial injury in rats. AB - OBJECTIVE: To assess alterations in endothelial barrier integrity and potential factors involved in zymosan-associated endothelial injury. DESIGN: Experimental study. SETTING: University hospital, Sweden. ANIMALS: 42 adult male Sprague Dawley rats. INTERVENTIONS: One hour before an intraperitoneal injection of paraffin or zymosan (0.25 mg/g body weight), 1.0 ml of a solution of saline, N acetyl-L-cysteine, dimethyl sulphoxide, indomethacin, verapamil, or allopurinol was given intravenously. MAIN OUTCOME MEASURES: Measurement of tissue water content, tissue intravascular plasma volume, interstitial fluid volume, and extravascular 125I-labelled human serum albumin distribution as well as plasma concentrations of albumin, alpha1-macroglobulin, alpha2-antiplasmin, and antithrombin III, 24 hours after the intraperitoneal injection. RESULTS: Endothelial permeability significantly increased in abdominal organs and the gastrointestinal tract, and plasma antiplasmin concentrations decreased. Pretreatment with N-acetyl-L-cysteine, dimethyl sulphoxide, or indomethacin protected against zymosan-induced endothelial barrier injury and the decline in protease inhibitors in plasma to varying degrees, while pretreatment with verapamil or allopurinol had a limited effect. CONCLUSION: Oxygen free radicals, prostaglandin, and proteases may have roles in the pathogenesis of zymosan induced endothelial barrier injuries, implying that several mediators probably are interacting. PMID- 10391169 TI - Effect of four enteral foods on the small bowel of undernourished rats after midgut resection. AB - OBJECTIVE: To describe the effects of new enteral foods on the adaptation of the gut mucosa after massive intestinal resection in rats. DESIGN: Laboratory experiment. SETTING: Teaching hospital, Spain. SUBJECTS: 91 male Wistar rats, 69 of which were studied (5 were excluded and 17 died). INTERVENTIONS: Previously undernourished rats were subjected to either massive bowel resection (n = 30) or laparotomy (n = 26) and fed four enteral hypocaloric diets for 7 days: Alitraq (n = 7 in each group), Impact (n = 8 and 7), Enrich (n = 8 and 6), and Elemental (n = 7 and 6). The remainder were not operated on and fed chow (n = 7) or a diet containing no protein (n = 6). Two diets were high in protein (Alitraq and Impact) and two contained normal amounts (Enrich and Elemental). MAIN OUTCOME MEASURES: Bowel mucosal thickness and proliferation; disaccharidase activity; intestinal weight and length; body weight; and plasma somatostatin, IGF-1, and peptide YY concentrations. RESULTS: Enriched diets provided a higher body and intestinal weight, and increased length of jejunal and ileal villous size. Peptide concentrations were modified by resection but not by the diet given. Concentrations of somatostatin and insulin-like growth factor were reduced in all groups with the exception of somatostatin in the two diets high in protein in the sham-operated rats. CONCLUSIONS: Enriched diets all improve the intestinal adaptive response to massive bowel resection in rats, offering advantages over diets with normal amounts of protein. PMID- 10391170 TI - Modified posterior preperitoneal mesh hernioplasty for repair of inguinal hernia. PMID- 10391171 TI - Acute abdomen caused by neutropenic enterocolitis: surgeon's dilemma. PMID- 10391172 TI - False aneurysm of an axillary artery in an intravenous drug misuser. PMID- 10391173 TI - Acute abdomen in mentally retarded patients: role of aerophagia. Report of nine cases. AB - Between 1993 and 1996 nine mentally retarded patients presented because of an acute abdomen. All had the habit of aerophagia, diagnosed previously by a general practitioner. Massive distension of the bowel led to ileus, volvulus, and necrosis. After placement of a percutaneous endoscopic gastrostomy catheter or performing a gastrostomy during laparotomy with the intention to use as a desufflator, no recurrence of the signs and symptoms of an acute abdomen were observed. PMID- 10391174 TI - Environmental tobacco smoke and ischaemic heart disease: a case study in applying causal criteria. AB - BACKGROUND: Whether ischaemic heart disease (IHD) is caused by exposure to environmental tobacco smoke (ETS), commonly known as "passive smoking", has been debated from both epidemiological and biological perspectives. METHODS AND RESULTS: In this paper we use Bradford Hill criteria to synthesize results from the biological and epidemiological literature in a formal assessment of the strength of support for such a relationship. Although we find that these criteria, designed for clinical trials, do not give an ideal framework for assessment of epidemiological and biological studies, nevertheless they do provide systematic guidance for this assessment. For the general population, of the nine tests proposed by Hill we find that one (biological plausibility) seems to be supported, though not unarguably; three (strength, consistency. specificity) appear to fail by accepted standards; and the remaining five have insufficient data for a clear evaluation (biological gradient, experimental evidence, temporality, coherence, analogy). Overall, this provides at best weak support for a causal association between ETS and IHD across the general community. Conversely, there appears to be more support, especially in the biology studies, for an association between ETS and IHD for those with preexisting disease, although epidemiological studies are limited in this area. CONCLUSIONS: One of the outcomes of this review is the identification of areas of focus for future epidemiological and biological research. First, we find that stronger associations may be found in the particular subpopulation with pre existing IHD. In this case, more convincing biological plausibility and experimental evidence indicate a need for relevant epidemiological studies, although individual responses are very variable. Second, we identify the need for further, more detailed evaluations of the nature of vessel wall thickenings occurring in experimental models of ETS exposure. Third, we propose long-term animal studies of initiation of IHD, including direct assessment of effects on the accumulation of lipid in vessel walls, at appropriate ETS exposure levels. PMID- 10391175 TI - Viremia in neonatal herpes simplex virus infections. AB - BACKGROUND: Polymerase chain reaction assays of the peripheral blood mononuclear cells (PBMC) and plasma may facilitate the diagnosis of neonatal herpes simplex virus (HSV). METHODS: Assays for HSV DNA were submitted from at least 1 specimen site (PBMC, plasma or cerebrospinal fluid) in 11 consecutive cases of neonatal HSV infection. RESULTS: HSV DNA was detected by PCR in the PBMC of 6 of 10 infants tested (60%), the plasma of 4 of 6 tested (67%) and the cerebrospinal fluid of 4 of 11 tested (36%). CONCLUSIONS: HSV viremia is more frequent than previously appreciated, and detection of HSV DNA in PBMC and plasma is a useful diagnostic tool, particularly in infants without skin lesions. PMID- 10391176 TI - Epiglottitis in Sweden before and after introduction of vaccination against Haemophilus influenzae type b. AB - BACKGROUND: Acute epiglottitis is an important manifestation of invasive Haemophilus influenzae type b (Hib) infection. In 1992 and 1993 Hib vaccination was introduced in the general childhood vaccination program in Sweden. The aim of the present investigation was to study the impact of Hib vaccination on the diagnosis of epiglottitis in Sweden in children as well as adults. METHODS: A retrospective national population-based study on the incidence of epiglottitis in Sweden was performed for the 10-year period 1987 to 1996. The incidence calculations were based on figures from the national register of all patients treated at Swedish hospitals. The incidence (cases/100,000/year) for the prevaccination period 1987 to 1991 was compared with the incidence after Hib vaccination was introduced. RESULTS: In children a substantial decrease was found after introduction of large scale vaccination against Hib. Below 5 years of age the annual incidence decreased from 20.9 in 1987 to 0.9 in 1996. In adults a tendency toward a decrease in incidence was evident. CONCLUSIONS: Introduction of Hib vaccination in a general childhood program was followed not only by a >90% reduction in the incidence in the youngest age group but also by a reduction in the incidence in the older age groups and among adults. PMID- 10391177 TI - Evaluation of young children in household contact with adult multidrug-resistant pulmonary tuberculosis cases. AB - BACKGROUND: The prevention and management of multidrug-resistant (MDR) tuberculosis has received much attention, but little attention has been given to children with MDR tuberculosis or children in contact with adults with MDR tuberculosis. The aim of this study was to determine the prevalence of tuberculous infection and disease in childhood contacts of adults with MDR pulmonary tuberculosis. METHOD: All children <5 years of age in household contact with 75 recently diagnosed adults with MDR pulmonary tuberculosis were evaluated. Evaluation included clinical examination, tuberculin skin test, chest radiography and culture for Mycobacterium tuberculosis from gastric aspirates. RESULTS: One hundred twenty-eight children, median age 27 months, were evaluated. Fifty children had recent contact with other adult tuberculosis cases. Sixty-six children previously had chemoprophylaxis or treatment of whom 36 defaulted treatment or received insufficient chemoprophylaxis. One child had HIV infection. Forty-seven children were classified as noninfected, 66 were considered infected only (Mantoux test, > or = 15 mm) and 15 had disease. Three children, who had not previously received antituberculosis drugs, had positive cultures for M. tuberculosis; all were multidrug-resistant. CONCLUSION: This study documents the transmission of multidrug-resistant M. tuberculosis to childhood contacts, the development of disease in these contacts and the importance of knowing the index case's M. tuberculosis susceptibility pattern in choosing a proper treatment regimen for the childhood contact. PMID- 10391178 TI - Population-based study of the incidence of Shigella diarrhea and causative serotypes in Santiago, Chile. AB - BACKGROUND: Shigella is an important cause of diarrheal disease in children in developing countries. The increasing prevalence of antibiotic-resistant strains has stimulated interest in the use of multivalent Shigella vaccines. Because Shigella vaccines under development are based on eliciting immunity to O antigens, monitoring the distribution of serotypes in defined target populations is critical. We initiated health center-based surveillance in a poor semirural community in Colina, Santiago (7489 children <60 months of age) to determine the age-specific incidence of Shigella disease and the responsible serotypes. FINDINGS: Surveillance was maintained at the 2 health centers during warm seasons (November 1 through April 30) for 4 successive years (1994 to 1998). Shigella was recovered from 54 of 243 cases of dysentery (22%) and from 215 of 3966 cases of nondysenteric diarrhea (5.4%) (P < 0.001). The peak mean annual incidence of shigellosis occurred among children 12 to 47 months of age (9.0 to 12.6 cases/10(3) children), although the incidence in infants (5.2/10(3)) and children 48 to 59 months of age (6.2/10(3)) was also substantial. During the 1995 through 1996 season, an age-matched healthy control was cultured for every child <60 months of age with diarrhea. Shigella isolation from cases (34 of 576, 5.9%) was >8-fold higher than controls (4 of 576, 0.7%) (P < 0.01). Four serotypes, Shigella sonnei (45%), Shigella flexneri 2b (19%), S. flexneri 2a (14%) and S. flexneri 6 (11%), accounted for 89% of all cases. INTERPRETATION: Shigella remains an important pediatric pathogen in Santiago. The serotype distribution from Colina, which closely resembles data from a population-based surveillance study in Santiago in the mid-1980s, demonstrates a remarkable degree of serotype stability in Santiago during a 15-year period. PMID- 10391179 TI - Treatment of severe pertussis: a study of the safety and pharmacology of intravenous pertussis immunoglobulin. AB - BACKGROUND: Pertussis in infants is often severe, resulting in complications and prolonged hospitalization. Treatment is limited to supportive care. Antibiotics do not significantly alter the course of the disease. Therapies directed at pertussis toxin, a major virulence factor of Bordetella pertussis, might be beneficial. This study examines the safety and pharmacology of intravenous pertussis immunoglobulin (P-IGIV), which has high levels of pertussis toxin antibodies. METHODS: P-IGIV was prepared as a 4% IgG solution from the pooled plasma from donors immunized with inactivated pertussis toxoid. The IgG pertussis toxin antibody concentration of 733 microg/ml is >7-fold higher than contained in conventional intravenous immunoglobulin products. Children with presumptive pertussis were allocated to one of three treatment doses of P-IGIV. RESULTS: Twenty-six of 30 enrolled children had confirmed pertussis. There were no adverse events associated with P-IGIV except one patient who had transient hypotension that responded to an infusion rate decrease. P-IGIV doses of 1500, 750 and 250 mg/kg achieved > or =4-fold, 3-fold and >2-fold rises in peak geometric mean titers of pertussis toxin IgG antibodies, respectively. P-IGIV exhibited a half life of 38.4 days and a volume of distribution of 87.8 ml/kg. All three treatment groups showed declines in lymphocytosis (P < 0.05) and paroxysmal coughing by the third day after P-IGIV infusion compared with preinfusion values. CONCLUSION: P IGIV is safe and achieves high pertussis toxin antibody titers in infants. This study provides data for a prospective, controlled trial of P-IGIV. PMID- 10391180 TI - Effect of changing antiretroviral therapy on human immunodeficiency virus viral load: experience with fifty-four perinatally infected children. AB - BACKGROUND: Experience in adults has shown that combination therapy including HIV protease inhibitors (PI) can profoundly affect viral replication and slow progression of HIV-associated disease. Trials defining the influence of PI and combination therapies on long term outcome of HIV infection in children have not yet been completed. Experience with infants and children who were receiving routine care in an HIV specialty clinic was reviewed to characterize the effect of changes involving one, two or three antiretrovirals. METHODS: Clinical and laboratory findings of children in whom antiretroviral therapy was changed were retrospectively reviewed. Successful response was defined as a reduction of viral load of at least 0.7 log10 RNA copies/ml lasting for at least 3 months. Differences in characteristics and the character of the response associated with successful and unsuccessful changes were analyzed. RESULTS: Of the 72 changes in therapy that were made in 54 children, 29 resulted in a successful response. A change involving 3 antiretrovirals was more likely to produce a successful response than a change involving 1 agent (6 of 9 vs. 6 of 24; P < 0.04). Reduction of viral load by > 100-fold or to undetectable amounts occurred more frequently in children who responded to a regimen containing a PI than in children who responded to reverse transcriptase inhibitors (11 of 21 vs. 1 of 8; P=0.05). Furthermore successful responses associated with addition of a PI were associated with a greater reduction in viral load than those that involved reverse transcriptase inhibitors (1.63+/-0.60 vs. 0.99+/-0.12 log10; P=0.003). CONCLUSIONS: This experience suggests that changing antiretroviral therapy in HIV infected children to regimens containing three drugs is more likely to result in a successful virologic outcome than changes in therapy involving one drug. This experience further supports the conclusion that including a PI as part of an antiretroviral regimen is more likely to result in a greater reduction in viral load in children. PMID- 10391181 TI - Risk factors for carriage of respiratory pathogens in the nasopharynx of healthy children. Ascanius Project Collaborative Group. AB - OBJECTIVES: To assess risk factors for nasopharyngeal carriage of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis in a large sample of healthy children. METHODS: In this point prevalence survey nasopharyngeal specimens were obtained from 1723 healthy children, ages 1 to 7 years, attending day-care centers or schools in 18 Italian cities. Written questionnaires for obtaining information about the demographics and medical history of the children were completed by the parents in the presence of a pediatrician. RESULTS: The overall carrier rate of respiratory pathogens was 17.9% (S. pneumoniae, 3.5%; H. influenzae, 11.9%; M. catarrhalis, 4.1%). Only 5% of S. pneumoniae strains were penicillin-resistant whereas approximately 40% were erythromycin-resistant. Beta lactamase production was found in 5.8% of H. influenzae and 88.7% of M. catarrhalis isolates. By multivariate analysis age (< or = 3 years), having older siblings, a history of prolonged full-time day-care attendance and living in a rural area significantly influenced the odds of carrying nasopharyngeal respiratory pathogens, particularly in children ages 1 to 5 years. Sex, breastfeeding, passive smoking and recent upper respiratory tract infections were not significant variables. Antibiotic treatment in the previous 3 months did not affect nasopharyngeal carriage, whereas repeated treatments with a macrolide were associated with carriage of S. pneumoniae. CONCLUSIONS: Our results suggest that there is a strong and long term relationship between exposure to large numbers of children in the first years of life and nasopharyngeal carriage of all respiratory pathogens. In addition antimicrobial restrictive guidelines should be tailored to local microbiologic sceneries. PMID- 10391182 TI - Intrapartum antibiotics and early onset neonatal sepsis caused by group B Streptococcus and by other organisms in Australia. Australasian Study Group for Neonatal Infections. AB - OBJECTIVE: Early onset group B streptococcal (EOGBS) infection, the major neonatal infection in industrialized countries, can be prevented by intrapartum antibiotics, but population studies are lacking. This study aimed to determine the incidence of early onset infections caused by group B Streptococcus (GBS) and other organisms in Australia and to assess intrapartum antibiotic use. DESIGN: Longitudinal, prospective surveillance of neonatal infections in Australian neonatal units from 1991 to 1997. Early onset infection defined as clinical sepsis in first 48 h after birth, with positive cultures of blood or cerebrospinal fluid or positive urine GBS antigen detection. RESULTS: The incidence of EOGBS sepsis fell from 2.0 per 1000 live births (95% confidence interval, 1.4, 2.5) in 1991 to 1993, to 1.3 (1.2, 1.4) in 1993 to 1995, to 0.5 (0.4, 0.7) in 1995 to 1997 (P < 0.0001). The incidence in Aboriginal babies was 5.2 (1.8, 8.6) in 1991 to 1993, 5.1 (3.0, 7.2) in 1993 to 1995 and 1.8 (1.1, 2.5) in 1995 to 1997 (P < 0.05). The incidence of early onset infections caused by organisms other than GBS also fell, from 1.2 per 1000 live births (0.8, 1.7) in 1991 to 1993, to 0.8 (0.7, 0.9) in 1993 to 1995 and 0.5 (0.3, 0.7) in 1995 to 1997 (P < 0.0001). In 1991, 3 of 9 study hospitals had a formal policy on intrapartum antibiotic use, whereas in 1997 all 11 hospitals had a formal policy (P=0.002). CONCLUSIONS: A steady fall in EOGBS infections in Australia from 1991 to 1997 has been associated with increasing use of intrapartum antibiotics. Increased antibiotic use is probably causal in the fall in GBS, because the incidence of early onset infections caused by other organisms has also fallen. PMID- 10391183 TI - Resource utilization associated with initial hospital stays complicated by early onset group B streptococcal disease. AB - BACKGROUND: The epidemiology of early onset neonatal group B streptococcal (GBS) disease has changed appreciably, but there are no recent assessments of the in hospital resource utilization it incurs. STUDY DESIGN: We performed a retrospective cohort study of infants delivered from 1987 through 1995 at Massachusetts' largest obstetrics hospital. A matched cohort design was used to assess care occurring after transfer to another acute care hospital. RESULTS: There were 135 cases of early onset neonatal GBS infection complicating 85,062 deliveries (1.6/1,000 births) in 9 years, with a substantial decline beginning in 1994, when maternal intrapartum chemoprophylaxis was widely introduced. Most (73%) infants had birth weights of 2500 g or more; 93% survived. Overall both the median and mean lengths of stay were 8 days longer for infants with GBS disease than for those without this infection (P < 0.001). Total hospital charges for neonates with GBS disease also were higher, with the difference in medians of $5323 and in means of $10,004 (P < 0.001). Differences were greatest among >2500 g birth weight infants; no excess was evident for infants with birth weights of < 1500 g. CONCLUSION: There was a substantial excess length of stay and charges associated with early onset neonatal GBS disease, although this was less than previously reported. PMID- 10391184 TI - Clinical utility of polymerase chain reaction testing for enteroviral meningitis. AB - BACKGROUND: During summer enteroviral meningitis is a common cause of febrile illness in children, who are typically hospitalized for 2 to 3 days if bacterial infection is suspected. It has been hypothesized that a sensitive polymerase chain reaction (PCR) assay could quickly confirm the diagnosis and subsequently decrease hospitalization costs. However, to have maximum impact results should be available within 24 h. This necessitates daily assays on small numbers of samples. METHODS: We examined the clinical utility of a PCR assay during two summers, comparing length of stay and charges. Only during the second summer were results reported to clinicians. Case controls were patients with negative PCR assay results but uncomplicated, presumed viral infections. We determined the cost per case identified with and without pleocytosis as a screen for PCR testing. RESULTS: During the first summer 25% (5/20) of patients with positive PCR assay results remained hospitalized for >2 days. During the second summer 10.2% (6 of 59) of children with positive enteroviral PCR assay results but 37.9% (25 of 66) of case controls remained hospitalized for >2 days. The mean length of hospitalization was significantly (P < 0.05) shorter for patients with positive PCR test results than for case controls. The material cost was approximately $238 per case identified. CONCLUSIONS: PCR testing has clinical utility for diagnosis of enteroviral meningitis. Although the demands for daily testing make the test expensive, it appears to be cost-effective with savings related to shorter hospital stays. PMID- 10391185 TI - Rocky Mountain spotted fever. PMID- 10391186 TI - Lymphocytic choriomeningitis virus: pediatric pathogen and fetal teratogen. PMID- 10391187 TI - Monoclonal antibodies against respiratory syncytial virus. PMID- 10391188 TI - Pneumococcal conjugate vaccines. PMID- 10391189 TI - Ehrlichial meningitis with cerebrospinal fluid morulae. PMID- 10391190 TI - Abdominal pain and eosinophilia in a ten-year-old boy. PMID- 10391191 TI - Peripheral vs. central blood cultures in patients admitted to a pediatric oncology ward. PMID- 10391192 TI - Vancomycin use in pediatric cardiothoracic surgery patients. PMID- 10391193 TI - How to increase by 40% the number of pediatric doses recovered from multidose vials of hepatitis B vaccines. PMID- 10391194 TI - Intrauterine transmission of coccidioidomycosis. PMID- 10391195 TI - Anaphylactic reaction to ciprofloxacin in a toddler: successful desensitization. PMID- 10391196 TI - Delayed myonecrosis in a leukemic patient with invasive group A streptococcal disease. PMID- 10391197 TI - Recurrent encephalopathy in cat-scratch disease. PMID- 10391198 TI - Sweet syndrome: an unusual presentation of chronic granulomatous disease in a child. PMID- 10391199 TI - Peritonitis caused by group B Streptococcus in a child receiving continuous cycling peritoneal dialysis. PMID- 10391200 TI - A three step approach to treating otitis media. PMID- 10391201 TI - Array data go public. PMID- 10391202 TI - Data analysis and integration: of steps and arrows. PMID- 10391203 TI - Architectural regulations and Hmg1. PMID- 10391204 TI - Clocks, criteria and critical genes. PMID- 10391205 TI - The weed paves the way. PMID- 10391206 TI - The candidate spermatogenesis gene RBMY has a homologue on the human X chromosome. PMID- 10391207 TI - RBMY evolved on the Y chromosome from a ubiquitously transcribed X-Y identical gene. PMID- 10391208 TI - Absence of Cd36 mutation in the original spontaneously hypertensive rats with insulin resistance. PMID- 10391209 TI - Characterization of single-nucleotide polymorphisms in coding regions of human genes. AB - A major goal in human genetics is to understand the role of common genetic variants in susceptibility to common diseases. This will require characterizing the nature of gene variation in human populations, assembling an extensive catalogue of single-nucleotide polymorphisms (SNPs) in candidate genes and performing association studies for particular diseases. At present, our knowledge of human gene variation remains rudimentary. Here we describe a systematic survey of SNPs in the coding regions of human genes. We identified SNPs in 106 genes relevant to cardiovascular disease, endocrinology and neuropsychiatry by screening an average of 114 independent alleles using 2 independent screening methods. To ensure high accuracy, all reported SNPs were confirmed by DNA sequencing. We identified 560 SNPs, including 392 coding-region SNPs (cSNPs) divided roughly equally between those causing synonymous and non-synonymous changes. We observed different rates of polymorphism among classes of sites within genes (non-coding, degenerate and non-degenerate) as well as between genes. The cSNPs most likely to influence disease, those that alter the amino acid sequence of the encoded protein, are found at a lower rate and with lower allele frequencies than silent substitutions. This likely reflects selection acting against deleterious alleles during human evolution. The lower allele frequency of missense cSNPs has implications for the compilation of a comprehensive catalogue, as well as for the subsequent application to disease association. PMID- 10391210 TI - Patterns of single-nucleotide polymorphisms in candidate genes for blood-pressure homeostasis. AB - Sequence variation in human genes is largely confined to single-nucleotide polymorphisms (SNPs) and is valuable in tests of association with common diseases and pharmacogenetic traits. We performed a systematic and comprehensive survey of molecular variation to assess the nature, pattern and frequency of SNPs in 75 candidate human genes for blood-pressure homeostasis and hypertension. We assayed 28 Mb (190 kb in 148 alleles) of genomic sequence, comprising the 5' and 3' untranslated regions (UTRs), introns and coding sequence of these genes, for sequence differences in individuals of African and Northern European descent using high-density variant detection arrays (VDAs). We identified 874 candidate human SNPs, of which 22% were confirmed by DNA sequencing to reveal a discordancy rate of 21% for VDA detection. The SNPs detected have an average minor allele frequency of 11%, and 387 are within the coding sequence (cSNPs). Of all cSNPs, 54% lead to a predicted change in the protein sequence, implying a high level of human protein diversity. These protein-altering SNPs are 38% of the total number of such SNPs expected, are more likely to be population-specific and are rarer in the human population, directly demonstrating the effects of natural selection on human genes. Overall, the degree of nucleotide polymorphism across these human genes, and orthologous great ape sequences, is highly variable and is correlated with the effects of functional conservation on gene sequences. PMID- 10391211 TI - Mutations in a gene encoding a new oxygen-regulated photoreceptor protein cause dominant retinitis pigmentosa. AB - The autosomal dominant retinitis pigmentosa (RP) locus, designated RP1, has been mapped through linkage studies to a 4-cM interval at 8q11-13. Here we describe a new photoreceptor-specific gene that maps in this interval and whose expression is modulated by retinal oxygen levels in vivo. This gene consists of at least 4 exons that encode a predicted protein of 2,156 amino acids. A nonsense mutation at codon 677 of this gene is present in approximately 3% of cases of dominant RP in North America. We also detected two deletion mutations that cause frameshifts and introduce premature termination codons in three other families with dominant RP. Our data suggest that mutations in this gene cause dominant RP, and that the encoded protein has an important but unknown role in photoreceptor biology. PMID- 10391212 TI - Mutations in a novel retina-specific gene cause autosomal dominant retinitis pigmentosa. AB - Inherited retinal diseases are a common cause of visual impairment in children and young adults, often resulting in severe loss of vision in later life. The most frequent form of inherited retinopathy is retinitis pigmentosa (RP), with an approximate incidence of 1 in 3,500 individuals worldwide. RP is characterized by night blindness and progressive degeneration of the midperipheral retina, accompanied by bone spicule-like pigmentary deposits and a reduced or absent electroretinogram (ERG). The disease process culminates in severe reduction of visual fields or blindness. RP is genetically heterogeneous, with autosomal dominant, autosomal recessive and X-linked forms. Here we have identified two mutations in a novel retina-specific gene from chromosome 8q that cause the RP1 form of autosomal dominant RP in three unrelated families. The protein encoded by this gene is 2,156 amino acids and its function is currently unknown, although the amino terminus has similarity to that of the doublecortin protein, whose gene (DCX) has been implicated in lissencephaly in humans. Two families have a nonsense mutation in codon 677 of this gene (Arg677stop), whereas the third family has a nonsense mutation in codon 679 (Gln679stop). In one family, two individuals homozygous for the mutant gene have more severe retinal disease compared with heterozygotes. PMID- 10391213 TI - Regulation of anterior/posterior patterning of the axial skeleton by growth/differentiation factor 11. AB - The bones that comprise the axial skeleton have distinct morphological features characteristic of their positions along the anterior/posterior axis. We previously described a novel TGF-beta family member, myostatin (encoded by the gene Mstn, formerly Gdf8), that has an essential role in regulating skeletal muscle mass. We also identified a gene related to Mstn by low-stringency screening. While the work described here was being completed, the cloning of this gene, designated Gdf11 (also called Bmp11), was also reported by other groups. Here we show that Gdf11, a new transforming growth factor beta(TGFbeta) superfamily member, has an important role in establishing this skeletal pattern. During early mouse embryogenesis, Gdf11 is expressed in the primitive streak and tail bud regions, which are sites where new mesodermal cells are generated. Homozygous mutant mice carrying a targeted deletion of Gdf11 exhibit anteriorly directed homeotic transformations throughout the axial skeleton and posterior displacement of the hindlimbs. The effect of the mutation is dose dependent, as Gdf11+/- mice have a milder phenotype than Gdf11-/- mice. Mutant embryos show alterations in patterns of Hox gene expression, suggesting that Gdf11 acts upstream of the Hox genes. Our findings suggest that Gdf11 is a secreted signal that acts globally to specify positional identity along the anterior/posterior axis. PMID- 10391214 TI - zA map for sequence analysis of the Arabidopsis thaliana genome. AB - Arabidopsis thaliana has emerged as a model system for studies of plant genetics and development, and its genome has been targeted for sequencing by an international consortium (the Arabidopsis Genome Initiative; http://genome-www. stanford.edu/Arabidopsis/agi.html). To support the genome-sequencing effort, we fingerprinted more than 20,000 BACs (ref. 2) from two high-quality publicly available libraries, generating an estimated 17-fold redundant coverage of the genome, and used the fingerprints to nucleate assembly of the data by computer. Subsequent manual revision of the assemblies resulted in the incorporation of 19,661 fingerprinted BACs into 169 ordered sets of overlapping clones ('contigs'), each containing at least 3 clones. These contigs are ideal for parallel selection of BACs for large-scale sequencing and have supported the generation of more than 5.8 Mb of finished genome sequence submitted to GenBank; analysis of the sequence has confirmed the integrity of contigs constructed using this fingerprint data. Placement of contigs onto chromosomes can now be performed, and is being pursued by groups involved in both sequencing and positional cloning studies. To our knowledge, these data provide the first example of whole-genome random BAC fingerprint analysis of a eucaryote, and have provided a model essential to efforts aimed at generating similar databases of fingerprint contigs to support sequencing of other complex genomes, including that of human. PMID- 10391215 TI - A complete BAC-based physical map of the Arabidopsis thaliana genome. AB - Arabidopsis thaliana is a small flowering plant that serves as the major model system in plant molecular genetics. The efforts of many scientists have produced genetic maps that provide extensive coverage of the genome (http://genome-www. stanford.edu/Arabidopsis/maps.html). Recently, detailed YAC, BAC, P1 and cosmid based physical maps (that is, representations of genomic regions as sets of overlapping clones of corresponding libraries) have been established that extend over wide genomic areas ranging from several hundreds of kilobases to entire chromosomes. These maps provide an entry to gain deeper insight into the A. thaliana genome structure. A. thaliana has been chosen as the subject of the first large-scale project intended to determine the full genome sequence of a plant. This sequencing project, together with the increasing interest in map based gene cloning, has highlighted the requirement for a complete and accurate physical map of this plant species. To supply the scientific community with a high-quality resource, we present here a complete physical map of A. thaliana using essentially the IGF BAC library. The map consists of 27 contigs that cover the entire genome, except for the presumptive centromeric regions, nucleolar organization regions (NOR) and telomeric areas. This is the first reported map of a complex organism based entirely on BAC clones and it represents the most homogeneous and complete physical map established to date for any plant genome. Furthermore, the analysis performed here serves as a model for an efficient physical mapping procedure using BAC clones that can be applied to other complex genomes. PMID- 10391216 TI - The lack of chromosomal protein Hmg1 does not disrupt cell growth but causes lethal hypoglycaemia in newborn mice. AB - High mobility group 1 (HMG1) protein is an abundant component of all mammalian nuclei, and related proteins exist in all eukaryotes. HMG1 binds linear DNA with moderate affinity and no sequence specificity, but bends the double helix significantly on binding through the minor groove. It binds with high affinity to DNA that is already sharply bent, such as linker DNA at the entry and exit of nucleosomes; thus, it is considered a structural protein of chromatin. HMG1 is also recruited to DNA by interactions with proteins required for basal and regulated transcriptions and V(D)J recombination. Here we generate mice harbouring deleted Hmg1. Hmg1-/- pups are born alive, but die within 24 hours due to hypoglycaemia. Hmg1-deficient mice survive for several days if given glucose parenterally, then waste away with pleiotropic defects (but no alteration in the immune repertoire). Cell lines lacking Hmg1 grow normally, but the activation of gene expression by the glucocorticoid receptor (GR, encoded by the gene Grl1) is impaired. Thus, Hmg1 is not essential for the overall organization of chromatin in the cell nucleus, but is critical for proper transcriptional control by specific transcription factors. PMID- 10391217 TI - Systematic determination of genetic network architecture. AB - Technologies to measure whole-genome mRNA abundances and methods to organize and display such data are emerging as valuable tools for systems-level exploration of transcriptional regulatory networks. For instance, it has been shown that mRNA data from 118 genes, measured at several time points in the developing hindbrain of mice, can be hierarchically clustered into various patterns (or 'waves') whose members tend to participate in common processes. We have previously shown that hierarchical clustering can group together genes whose cis-regulatory elements are bound by the same proteins in vivo. Hierarchical clustering has also been used to organize genes into hierarchical dendograms on the basis of their expression across multiple growth conditions. The application of Fourier analysis to synchronized yeast mRNA expression data has identified cell-cycle periodic genes, many of which have expected cis-regulatory elements. Here we apply a systematic set of statistical algorithms, based on whole-genome mRNA data, partitional clustering and motif discovery, to identify transcriptional regulatory sub-networks in yeast-without any a priori knowledge of their structure or any assumptions about their dynamics. This approach uncovered new regulons (sets of co-regulated genes) and their putative cis-regulatory elements. We used statistical characterization of known regulons and motifs to derive criteria by which we infer the biological significance of newly discovered regulons and motifs. Our approach holds promise for the rapid elucidation of genetic network architecture in sequenced organisms in which little biology is known. PMID- 10391218 TI - Mutations in a delta 8-delta 7 sterol isomerase in the tattered mouse and X linked dominant chondrodysplasia punctata. jderry@immunex.com. AB - Tattered (Td) is an X-linked, semi-dominant mouse mutation associated with prenatal male lethality. Heterozygous females are small and at 4-5 days of age develop patches of hyperkeratotic skin where no hair grows, resulting in a striping of the coat in adults. Craniofacial anomalies and twisted toes have also been observed in some affected females. A potential second allele of Td has also been described. The phenotype of Td is similar to that seen in heterozygous females with human X-linked dominant chondrodysplasia punctata (CDPX2, alternatively known as X-linked dominant Conradi-Hunermann-Happle syndrome) as well as another X-linked, semi-dominant mouse mutation, bare patches (Bpa). The Bpa gene has recently been identified and encodes a protein with homology to 3beta-hydroxysteroid dehydrogenases that functions in one of the later steps of cholesterol biosynthesis. CDPX2 patients display skin defects including linear or whorled atrophic and pigmentary lesions, striated hyperkeratosis, coarse lusterless hair and alopecia, cataracts and skeletal abnormalities including short stature, rhizomelic shortening of the limbs, epiphyseal stippling and craniofacial defects (MIM 302960). We have now identified the defect in Td mice as a single amino acid substitution in the delta8-delta7 sterol isomerase emopamil binding protein (Ebp; encoded by Ebp in mouse) and identified alterations in human EBP in seven unrelated CDPX2 patients. PMID- 10391219 TI - Mutations in the gene encoding 3 beta-hydroxysteroid-delta 8, delta 7-isomerase cause X-linked dominant Conradi-Hunermann syndrome. AB - X-linked dominant Conradi-Hunermann syndrome (CDPX2; MIM 302960) is one of a group of disorders with aberrant punctate calcification in cartilage, or chondrodysplasia punctata (CDP). This is most prominent around the vertebral column, pelvis and long bones in CPDX2. Additionally, CDPX2 patients may have asymmetric rhizomesomelia, sectorial cataracts, patchy alopecia, ichthyosis and atrophoderma. The phenotype in CDPX2 females ranges from stillborn to mildly affected individuals identified in adulthood. CDPX2 is presumed lethal in males, although a few affected males have been reported. We found increased 8(9) cholestenol and 8-dehydrocholesterol in tissue samples from seven female probands with CDPX2 (ref. 4). This pattern of accumulated cholesterol intermediates suggested a deficiency of 3beta-hydroxysteroid-delta8,delta7-isomerase (sterol delta8-isomerase), which catalyses an intermediate step in the conversion of lanosterol to cholesterol. A candidate gene encoding a sterol-delta8-isomerase (EBP) has been identified and mapped to Xp11.22-p11.23 (refs 5,6). Using SSCP analysis and sequencing of genomic DNA, we found EBP mutations in all probands. We confirmed the functional significance of two missense alleles by expressing them in a sterol-delta8-isomerase-deficient yeast strain. Our results indicate that defects in sterol-delta8-isomerase cause CDPX2 and suggest a role for sterols in bone development. PMID- 10391220 TI - Cryptorchidism in mice mutant for Insl3. AB - Impaired testicular descent (cryptorchidism) is one of the most frequent congenital abnormalities in humans, involving 2% of male births. Cryptorchidism can result in infertility and increases risk for development of germ-cell tumours. Testicular descent from abdomen to scrotum occurs in two distinct phases: the trans-abdominal phase and the inguino-scrotal phase. Currently, little is known about the factors that regulate the trans-abdominal phase of testicular descent. Leydig insulin-like hormone (Insl3) is a member of the insulin hormone superfamily expressed in the developing testis. We show here that mice mutant for Insl3 are viable, but exhibit bilateral cryptorchidism due to developmental abnormalities of the gubernaculum, resulting in abnormal spermatogenesis and infertility. Female homozygotes have impaired fertility associated with deregulation of the oestrus cycle. These findings reveal roles for Insl3 in the development of the urogenital tract and in female fertility. Insl3 may act as a hormone to regulate the growth and differentiation of the gubernaculum, thereby mediating intra-abdominal testicular descent. PMID- 10391221 TI - Mutations in SLC19A2 cause thiamine-responsive megaloblastic anaemia associated with diabetes mellitus and deafness. AB - Thiamine-responsive megaloblastic anaemia (TRMA), also known as Rogers syndrome, is an early onset, autosomal recessive disorder defined by the occurrence of megaloblastic anaemia, diabetes mellitus and sensorineural deafness, responding in varying degrees to thiamine treatment (MIM 249270). We have previously narrowed the TRMA locus from a 16-cM to a 4-cM interval on chromosomal region 1q23.3 (refs 3,4) and this region has been further refined to a 1.4-cM interval. Previous studies have suggested that deficiency in a high-affinity thiamine transporter may cause this disorder. Here we identify the TRMA gene by positional cloning. We assembled a P1-derived artificial chromosome (PAC) contig spanning the TRMA candidate region. This clarified the order of genetic markers across the TRMA locus, provided 9 new polymorphic markers and narrowed the locus to an approximately 400-kb region. Mutations in a new gene, SLC19A2, encoding a putative transmembrane protein homologous to the reduced folate carrier proteins, were found in all affected individuals in six TRMA families, suggesting that a defective thiamine transporter protein (THTR-1) may underlie the TRMA syndrome. PMID- 10391222 TI - The gene mutated in thiamine-responsive anaemia with diabetes and deafness (TRMA) encodes a functional thiamine transporter. AB - Thiamine-responsive megaloblastic anaemia with diabetes and deafness (TRMA; MIM 249270) is an autosomal recessive disease thought to be due to a defect in thiamine (vitamin B1) transport. Pharmacological doses of thiamine correct the anaemia, and in some cases improve the diabetes, although progressive sensorineural deafness is irreversible. Previous studies localized the TRMA gene to a 4-cM region on chromosome 1q23.3 (ref. 5), and fine-mapping has recently narrowed that region further. We have previously demonstrated that fibroblasts from people with TRMA lack high-affinity thiamine transport. Expression of a gene encoding a known yeast thiamine transporter, THI10 (refs 8-10), in TRMA mutant cells prevents apoptotic cell death in thiamine-depleted medium. On the basis of these studies, we hypothesized that a defective thiamine transporter causes TRMA. We undertook a candidate gene approach to identify putative thiamine transporters in the 1q23.3 critical region. Here we present evidence that the gene SLC19A2 (for solute carrier family 19 (thiamine transporter), member 2) encodes the first known mammalian thiamine transporter, which we designate thiamine transporter-1 (THTR-1). PMID- 10391224 TI - Time to grasp the international perspective on GM crops. PMID- 10391223 TI - Mutations in a new gene encoding a thiamine transporter cause thiamine-responsive megaloblastic anaemia syndrome. AB - Thiamine-responsive megaloblastic anaemia syndrome (TRMA; MIM 249270) is an autosomal recessive disorder with features that include megaloblastic anaemia, mild thrombocytopenia and leucopenia, sensorineural deafness and diabetes mellitus. Treatment with pharmacologic doses of thiamine ameliorates the megaloblastic anaemia and diabetes mellitus. A defect in the plasma membrane transport of thiamine has been demonstrated in erythrocytes and cultured skin fibroblasts from TRMA patients. The gene causing TRMA was assigned to 1q23.2 q23.3 by linkage analysis. Here we report the cloning of a new gene, SLC19A2, identified from high-through-put genomic sequences due to homology with SLC19A1, encoding reduced folate carrier 1 (refs 8-10). We cloned the entire coding region by screening a human fetal brain cDNA library. SLC19A2 encodes a protein (of 497 aa) predicted to have 12 transmembrane domains. We identified 2 frameshift mutations in exon 2. a 1-bp insertion and a 2-bp deletion, among four Iranian families with TRMA. The sequence homology and predicted structure of SLC19A2, as well as its role in TRMA, suggest that its gene product is a thiamine carrier, the first to be identified in complex eukaryotes. PMID- 10391225 TI - Gore under fire in controversy over South Africa AIDS drug law. PMID- 10391226 TI - Chlorine industry says EPA rules ignore good science. PMID- 10391227 TI - Japan tightens rules on GM crops to protect the environment. PMID- 10391228 TI - Varmus defends plan for global biomedical e-journal. PMID- 10391229 TI - One-stop shop for 200 life science journals. PMID- 10391230 TI - Israel 'must relax technology transfer law'. PMID- 10391231 TI - Researcher fights suspension over funds. PMID- 10391232 TI - Bioethicists must come down to Earth. PMID- 10391233 TI - German researchers won't be put in the dock. PMID- 10391234 TI - Hox clusters. Size doesn't matter. PMID- 10391235 TI - DNA damage enables p73. PMID- 10391236 TI - Evolutionary genetics. No sex please, we're fungi. PMID- 10391237 TI - Alzheimer's disease. Pinning down phosphorylated tau. PMID- 10391238 TI - Menstrual cycle alters face preference. PMID- 10391239 TI - Limbs move beyond the radical fringe. PMID- 10391240 TI - A reversibly antigen-responsive hydrogel. AB - Stimuli-responsive hydrogels that undergo abrupt changes in volume in response to external stimuli such as pH, temperature and solvent composition have potential applications in biomedicine and the creation of 'intelligent' materials systems, for example as media for drug delivery, separation processes and protein immobilization. Hydrogels have been reported that respond to pH, temperature, electric fields and saccharides. For some biomedical applications it would be very useful to have a material whose swelling response was dictated by a specific protein. Here we report such a material, which swells reversibly in a buffer solution in response to a specific antigen. The hydrogel was prepared by grafting the antigen and corresponding antibody to the polymer network, so that binding between the two introduces crosslinks in the network. Competitive binding of the free antigen triggers a change in gel volume owing to breaking of these non covalent crosslinks. In addition, we show that the hydrogel displays shape-memory behaviour, and that stepwise changes in antigen concentration can induce pulsatile permeation of a protein through the network. PMID- 10391241 TI - Hox genes in brachiopods and priapulids and protostome evolution. AB - Understanding the early evolution of animal body plans requires knowledge both of metazoan phylogeny and of the genetic and developmental changes involved in the emergence of particular forms. Recent 18S ribosomal RNA phylogenies suggest a three-branched tree for the Bilateria comprising the deuterostomes and two great protostome clades, the lophotrochozoans and ecdysozoans. Here, we show that the complement of Hox genes in critical protostome phyla reflects these phylogenetic relationships and reveals the early evolution of developmental regulatory potential in bilaterians. We have identified Hox genes that are shared by subsets of protostome phyla. These include a diverged pair of posterior (Abdominal-B like) genes in both a brachiopod and a polychaete annelid, which supports the lophotrochozoan assemblage, and a distinct posterior Hox gene shared by a priapulid, a nematode and the arthropods, which supports the ecdysozoan clade. The ancestors of each of these two major protostome lineages had a minimum of eight to ten Hox genes. The major period of Hox gene expansion and diversification thus occurred before the radiation of each of the three great bilaterian clades. PMID- 10391242 TI - A stop-codon mutation in the BRI gene associated with familial British dementia. AB - Familial British dementia (FBD), previously designated familial cerebral amyloid angiopathy-British type, is an autosomal dominant disorder of undetermined origin characterized by progressive dementia, spasticity, and cerebellar ataxia, with onset at around the fifth decade of life. Cerebral amyloid angiopathy, non neuritic and perivascular plaques and neurofibrillary tangles are the predominant pathological lesions. Here we report the identification of a unique 4K protein subunit named ABri from isolated amyloid fibrils. This highly insoluble peptide is a fragment of a putative type-II single-spanning transmembrane precursor that is encoded by a novel gene, BRI, located on chromosome 13. A single base substitution at the stop codon of this gene generates a longer open reading frame, resulting in a larger, 277-residue precursor. Release of the 34 carboxy terminal amino acids from the mutated precursor generates the ABri amyloid subunit. The mutation creates a cutting site for the restriction enzyme XbaI, which is useful for detecting asymptomatic carriers. Antibodies against the amyloid or homologous synthetic peptides recognize both parenchymal and vascular lesions in FBD patients. A point mutation at the stop codon of BRI therefore results in the generation of the ABri peptide, which is deposited as amyloid fibrils causing neuronal disfunction and dementia. PMID- 10391243 TI - Human theta oscillations exhibit task dependence during virtual maze navigation. AB - Theta oscillations (electroencephalographic activity with a frequency of 4-8 Hz) have long been implicated in spatial navigation in rodents; however, the role of theta oscillators in human spatial navigation has not been explored. Here we describe subdural recordings from epileptic patients learning to navigate computer-generated mazes. Visual inspection of the raw intracranial signal revealed striking episodes of high-amplitude slow-wave oscillations at a number of areas of the cortex, including temporal cortex. Spectral analysis showed that these oscillations were in the theta band. These episodes of theta activity, which typically last several cycles, are dependent on task characteristics. Theta oscillations occur more frequently in more complex mazes; they are also more frequent during recall trials than during learning trials. PMID- 10391244 TI - The prolyl isomerase Pin1 restores the function of Alzheimer-associated phosphorylated tau protein. AB - One of the neuropathological hallmarks of Alzheimer's disease is the neurofibrillary tangle, which contains paired helical filaments (PHFs) composed of the microtubule-associated protein tau. Tau is hyperphosphorylated in PHFs, and phosphorylation of tau abolishes its ability to bind microtubules and promote microtubule assembly. Restoring the function of phosphorylated tau might prevent or reverse PHF formation in Alzheimer's disease. Phosphorylation on a serine or threonine that precedes proline (pS/T-P) alters the rate of prolyl isomerization and creates a binding site for the WW domain of the prolyl isomerase Pin1. Pin1 specifically isomerizes pS/T-P bonds and regulates the function of mitotic phosphoproteins. Here we show that Pin1 binds to only one pT-P motif in tau and copurifies with PHFs, resulting in depletion of soluble Pin1 in the brains of Alzheimer's disease patients. Pin1 can restore the ability of phosphorylated tau to bind microtubules and promote microtubule assembly in vitro. As depletion of Pin1 induces mitotic arrest and apoptotic cell death, sequestration of Pin1 into PHFs may contribute to neuronal death. These findings provide a new insight into the pathogenesis of Alzheimer's disease. PMID- 10391245 TI - Characterization of the human cysteinyl leukotriene CysLT1 receptor. AB - The cysteinyl leukotrienes-leukotriene C4(LTC4), leukotriene D4(LTD4) and leukotriene E4(LTE4)-are important mediators of human bronchial asthma. Pharmacological studies have determined that cysteinyl leukotrienes activate at least two receptors, designated CysLT1 and CysLT2. The CysLT1-selective antagonists, such as montelukast (Singulair), zafirlukast (Accolate) and pranlukast (Onon), are important in the treatment of asthma. Previous biochemical characterization of CysLT1 antagonists and the CysLT1 receptor has been in membrane preparations from tissues enriched for this receptor. Here we report the molecular and pharmacological characterization of the cloned human CysLT1 receptor. We describe the functional activation (calcium mobilization) of this receptor by LTD4 and LTC4, and competition for radiolabelled LTD4 binding to this receptor by the cysteinyl leukotrienes and three structurally distinct classes of CysLT1-receptor antagonists. We detected CysLT1-receptor messenger RNA in spleen, peripheral blood leukocytes and lung. In normal human lung, expression of the CysLT1-receptor mRNA was confined to smooth muscle cells and tissue macrophages. Finally, we mapped the human CysLT1-receptor gene to the X chromosome. PMID- 10391246 TI - MAP kinase and Wnt pathways converge to downregulate an HMG-domain repressor in Caenorhabditis elegans. AB - The signalling protein Wnt regulates transcription factors containing high mobility-group (HMG) domains to direct decisions on cell fate during animal development. In Caenorhabditis elegans, the HMG-domain-containing repressor POP-1 distinguishes the fates of anterior daughter cells from their posterior sisters throughout development, and Wnt signalling downregulates POP-1 activity in one posterior daughter cell called E. Here we show that the genes mom-4 and lit-1 are also required to downregulate POP-1, not only in E but also in other posterior daughter cells. Consistent with action in a common pathway, mom-4 and lit-1 exhibit similar mutant phenotypes and encode components of the mitogen-activated protein kinase (MAPK) pathway that are homologous to vertebrate transforming growth-factor-beta-activated kinase (TAK1) and NEMO-like kinase (NLK), respectively. Furthermore, MOM-4 and TAK1 bind related proteins that promote their kinase activities. We conclude that a MAPK-related pathway cooperates with Wnt signal transduction to downregulate POP-1 activity. These functions are likely to be conserved in vertebrates, as TAK1 and NLK can downregulate HMG domain-containing proteins related to POP-1. PMID- 10391247 TI - The TAK1-NLK-MAPK-related pathway antagonizes signalling between beta-catenin and transcription factor TCF. AB - The Wnt signalling pathway regulates many developmental processes through a complex of beta-catenin and the T-cell factor/lymphoid enhancer factor (TCF/LEF) family of high-mobility-group transcription factors. Wnt stabilizes cytosolic beta-catenin, which then binds to TCF and activates gene transcription. This signalling cascade is conserved in vertebrates, Drosophila and Caenorhabditis elegans. In C. elegans, the proteins MOM-4 and LIT-1 regulate Wnt signalling to polarize responding cells during embryogenesis. MOM-4 and LIT-1 are homologous to TAK1 (a kinase activated by transforming growth factor-beta) mitogen-activated protein-kinase-kinase kinase (MAP3K) and MAP kinase (MAPK)-related NEMO-like kinase (NLK), respectively, in mammalian cells. These results raise the possibility that TAK1 and NLK are also involved in Wnt signalling in mammalian cells. Here we show that TAK1 activation stimulates NLK activity and downregulates transcriptional activation mediated by beta-catenin and TCF. Injection of NLK suppresses the induction of axis duplication by microinjected beta-catenin in Xenopus embryos. NLK phosphorylates TCF/LEF factors and inhibits the interaction of the beta-catenin-TCF complex with DNA. Thus, the TAK1-NLK-MAPK like pathway negatively regulates the Wnt signalling pathway. PMID- 10391248 TI - TRA-1 regulates the cellular distribution of the tra-2 mRNA in C. elegans. AB - The GLI protein family is involved in several key developmental processes in both vertebrates and invertebrates. The Drosophila GLI protein, Cubitus interuptus (Ci), regulates segment polarity and wing and leg development. In vertebrates, the GLI proteins control neural, lung, bone and gut development. In the nematode Caenorhabditis elegans, the GLI family member TRA-1 is necessary for normal sexual development. GLI, Ci and TRA-1 each contain five zinc-finger domains and bind the identical DNA sequence. Previous analyses are consistent with these proteins being transcription factors. Here we show that TRA-1 can act posttranscriptionally to govern gene activity. Our results indicate that the binding of TRA-1 to the 3' untranslated region of tra-2 regulates the export of tra-2 messenger RNA from the nucleus. The fact that TRA-1 is part of a conserved family of proteins raises the possibility that GLI family members are both transcriptional and post-transcriptional regulators of gene expression. PMID- 10391249 TI - The tyrosine kinase c-Abl regulates p73 in apoptotic response to cisplatin induced DNA damage. AB - Cancer chemotherapeutic agents such as cisplatin exert their cytotoxic effect by inducing DNA damage and activating programmed cell death (apoptosis). The tumour suppressor protein p53 is an important activator of apoptosis. Although p53 deficient cancer cells are less responsive to chemotherapy, their resistance is not complete, which suggests that other apoptotic pathways may exist. A p53 related gene, p73, which encodes several proteins as a result of alternative splicing, can also induce apoptosis. Here we show that the amount of p73 protein in the cell is increased by cisplatin. This induction of p73 is not seen in cells unable to carry out mismatch repair and in which the nuclear enzyme c-Abl tyrosine kinase is not activated by cisplatin. The half-life of p73 is prolonged by cisplatin and by co-expression with c-Abl tyrosine kinase; the apoptosis inducing function of p73 is also enhanced by the c-Abl kinase. Mouse embryo fibroblasts deficient in mismatch repair or in c-Abl do not upregulate p73 and are more resistant to killing by cisplatin. Our results indicate that c-Abl and p73 are components of a mismatch-repair-dependent apoptosis pathway which contributes to cisplatin-induced cytotoxicity. PMID- 10391250 TI - Interaction of c-Abl and p73alpha and their collaboration to induce apoptosis. AB - c-Abl, a non-receptor tyrosine kinase, is activated by agents that damage DNA. This activation results in either arrest of the cell cycle in phase G1 or apoptotic cell death, both of which are dependent on the kinase activity of c Abl. p73, a member of the p53 family of tumour-suppressor proteins, can also induce apoptosis. Here we show that the apoptotic activity of p73alpha requires the presence of functional, kinase-competent c-Abl. Furthermore, p73 and c-Abl can associate with each other, andthis binding is mediated by a PxxP motif in p73 and the SH3 domain of c-Abl. We find that p73 is a substrate of the c-Abl kinase and that the ability of c-Abl to phosphorylate p73 is markedly increased by gamma irradiation. Moreover, p73 is phosphorylated in vivo in response to ionizing radiation. These findings define a pro-apoptotic signalling pathway involving p73 and c-Abl. PMID- 10391251 TI - p73 is regulated by tyrosine kinase c-Abl in the apoptotic response to DNA damage. AB - The protein p73 is a structural and functional homologue of the p53 tumour suppressor protein but, unlike p53, it is not induced in response to DNA damage. The tyrosine kinase c-Abl is activated by certain DNA-damaging agents and contributes to the induction of programmed cell death (apoptosis) by p53 dependent and p53-independent mechanisms. Here we show that c-Abl binds to p73 in cells, interacting through its SH3 domain with the carboxy-terminal homo oligomerization domain of p73. c-Abl phosphorylates p73 on a tyrosine residue at position 99 both in vitro and in cells that have been exposed to ionizing radiation. Our results show that c-Abl stimulates p73-mediated transactivation and apoptosis. This regulation of p73 by c-Abl in response to DNA damage is also demonstrated by a failure of ionizing-radiation-induced apoptosis after disruption of the c-Abl-p73 interaction. These findings show that p73 is regulated by a c-Abl-dependent mechanism and that p73 participates in the apoptotic response to DNA damage. PMID- 10391252 TI - Informed advice. PMID- 10391253 TI - Influence of cardiopulmonary bypass perfusion temperature on neurologic and hematologic function after coronary artery bypass grafting. AB - BACKGROUND AND METHODS: A National Institutes of Health-sponsored trial (1994 to 1998) randomized patients undergoing coronary artery bypass grafting that required three or more grafts to receive perfusion at either cold (20 degrees C), tepid (32 degrees C), or warm (37 degrees C) temperature. The goal of the study was to evaluate morbidity, primarily neurologic dysfunction and secondarily hematologic factors. One thousand seven hundred seventy-seven patients were screened and 291 enrolled. Neurologic function was studied by a dedicated pool of blinded neurologists. A standard test battery termed the Mathew Scale using three subscales--cognitive function, elemental skills, and disability--was used to study central nervous system function. Hematologic function was assessed in 53 of the 291 patients with measurements of postoperative fibrinolytic potential. RESULTS: All preoperative and operative data were comparable between groups. A decrease in Mathew Scale was seen in 69% of patient from before operation to immediately after operation. However, between the early postoperative study and the 1-month follow-up, 48% of patients had returned to baseline. There was no difference noted across temperature groups in any neurologic parameter of function. In all, 55% of the group were at or above their preoperative level at 1 month. Forty-nine patients suspect for cerebrovascular accident had a computed tomographic scan, but only 13 (4.5%) had a documented cerebrovascular accident (4 patients in the warm, 3 in the tepid, and 6 patients in the cold group). Fibrinolytic changes correlated with perfusion temperature documented that fibrinolysis was most active at 37 degrees C. Thus, increasing perfusate temperature increases fibrinolysis, which was associated with reoperation for bleeding in 4% warm group patients, 1% tepid, and 0% cold group patients (0.1 > p > 0.05). No other perioperative complications were temperature related. There were 4 deaths (1.4%) (1 in the warm group, 2 in the tepid group, and 1 in the cold group). CONCLUSIONS: (1) Persistent postoperative neurologic dysfunction at 1 month occurs in 36% of patients undergoing coronary artery bypass grafting and is not related to a cerebrovascular accident; 2) perfusion temperature has no relationship to neurologic function after bypass; and 3) fibrinolytic activity is greatest at warm temperatures. PMID- 10391254 TI - Favorable results after sleeve lobectomy or bronchoplasty for bronchial malignancies. AB - BACKGROUND: Sleeve lobectomy and bronchoplasty are established alternatives to pneumonectomy for bronchial malignancies involving a main bronchus. However, potential bronchial anastomotic complications have deterred the general application of these types of resection. Some reports have contained a mixture of non-small cell lung cancer (NSCLC) and tumors of low-grade malignancy, making it difficult to assess the long-term results of these procedures as an alternative to pneumonectomy for lung cancer. METHODS: We retrospectively reviewed our experience with sleeve lobectomy and bronchoplasty for bronchial malignancies from January 1988 to September 1998 separating NSCLC (n = 58) from tumors of low grade malignancy (n = 19). We compared the overall results between sleeve lobectomy and pneumonectomy (n = 142) performed for NSCLC over the same time interval. RESULTS: For NSCLC, after sleeve lobectomy, the operative mortality was 5.2% (3 of 58 patients) and the overall 5-year actuarial survival was 37.5%. After pneumonectomy, the operative mortality was 4.9% (7 of 142 patients) and the overall 5-year actuarial survival was 35.8%. For tumors with low-grade malignancy, there was no operative mortality after sleeve lobectomy or bronchoplasty and the 5-year actuarial survival was 100%. Major bronchial anastomotic complications occurred in 3 patients among the 77 patients who underwent sleeve resection. CONCLUSIONS: Sleeve resection can be performed with a low risk of bronchial anastomotic complication. The long-term survival after sleeve resection for NSCLC is similar to pneumonectomy. Excellent results are obtained after sleeve resection for low-grade malignancies. PMID- 10391255 TI - Efficacy and safety of extended thymectomy for elderly patients with myasthenia gravis. AB - BACKGROUND: The number of elderly patients who are diagnosed as myasthenia gravis (MG) is increasing in Japan. Although several factors affecting thymectomy have been well documented, few studies have focused on the efficacy and safety of thymectomy for elderly patients older than 60 years. METHODS: We evaluated 94 patients with MG who underwent extended thymectomy, and divided them into two groups: patients younger than 59 years and patients older than 60 years. Preoperative patient data, pathology of the thymus, complications, and clinical outcome were evaluated. RESULTS: In 69 young patients and 25 elderly patients, we observed no significant differences between the two groups with regard to preoperative data. Thymic hyperplasia was present in 45% of the young group and 16% of the elderly group. Remission and improvement rate were 40% and 57% in the young group and 8% and 75% in the elderly group, respectively. There were no serious complications, except one early death due to gastrointestinal bleeding in the elderly group. CONCLUSIONS: We conclude that thymectomy is a safe and effective alternative for elderly patients with MG. PMID- 10391256 TI - Prognostic factors for myasthenia gravis treated by thymectomy: review of 61 cases. AB - BACKGROUND: Medical treatment for myasthenia gravis (MG) involves the use of anticholinesterase agents, immunosuppressive drugs, plasmapheresis, and gamma globulin. However, these agents result in a complete clinical remission rate as low as 15%. As a consequence, thymectomy, preferably by transsternal approach, has become increasingly accepted as an efficacious procedure for MG, with reported complete clinical remission rates as high as 80%. METHODS: We have the clinical records of 61 patients diagnosed with MG at La Paz University Hospital, Madrid, Spain, from January 1977 to December 1994. All patients underwent thymectomy. The purpose of this investigation was to determine the major prognostic factors predicting MG outcome after operation. RESULTS: Our results indicate that patients with a length of the disease from onset to operation shorter than 8 months have the best prognosis. Ossermann stages I and III are also associated with higher complete clinical remission rates. In contrast, neither age nor sex were found to be significantly related to MG outcome after thymectomy, although female patients have better prognosis than men, and the younger the patient the more likely is complete clinical remission. Pathologic findings after the operation were not found to be of prognostic value either. CONCLUSIONS: We conclude that thymectomy is a beneficial procedure for MG patients, with a complete clinical remission rate of 46% at 5 years postoperatively in our series. Therefore we advocate thymectomy for MG patients as early as possible in the course of disease because time elapsed from diagnosis to operation is the main determinant of the outcome. PMID- 10391257 TI - Prognostic significance of surgical-pathologic N1 disease in non-small cell carcinoma of the lung. AB - BACKGROUND: N1 disease represents a heterogeneous group of non-small cell lung carcinoma with varying 5-year survival rates. Specific types of N1 lymph node involvement need to be further investigated and their prognostic significance clarified. METHODS: From 1984 to 1993, 1,174 patients with non-small cell lung cancer had complete mediastinal lymph node dissection: N0, 50.25% (n = 590); N1, 21.8% (n = 256); and N2, 27.95% (n = 328). The N1 subgroup cases were reviewed. Four levels of N1 nodes were identified using the New Regional Lymph Node Classification for Lung Cancer Staging. Their prognostic significances were tested and 5-year survival rates were compared with those of N0 and N2 patients of the whole group. RESULTS: The overall 5-year survival rate of N1 patients was 47.5%. Survival was not related to site of the primary lung cancer, pathologic T factor, histologic type, type of resection, number of N1 station involved, nor type of N1 involvement (direct extension or metastases). Five-year survival was significantly better when N1 involvement was intralobar (levels 12 and 13, n = 102), as compared with extralobar (hilar) involvement (levels 10 and 11, n = 154): 53.6% versus 38.5% (p = 0.02). Intralobar N1 5-year survival was similar to that of N0 (53.6% vs 56.5%, p = 0.01), and extralobar 5-year survival with that of N2 (38.5 vs 28.3%, p = 0.01) when N2 was present in only one station in the ipsilateral mediastinum. CONCLUSIONS: N1 disease is a compound of two subgroups: one located inside the lobes is related to N0, and the other (extralobar or hilar) behaves like an early stage of N2 disease. This offers further information for clinical, therapeutic, and research purposes. PMID- 10391258 TI - Impact of graft ischemic time on outcomes after bilateral sequential single-lung transplantation. AB - BACKGROUND: Graft ischemic time (GIT) is a potential limiting factor in lung transplantation. METHODS: Seventy-four patients who underwent bilateral sequential single-lung transplantation were divided into three groups: group I, GIT less than 5 hours (n = 20); group II, GIT between 5 and 8 hours (n = 39); and group III, GIT more than 8 hours (n = 15). We compared early allograft function (ratio of arterial oxygen tension to inspired oxygen fraction and alveolar arterial oxygen gradient), blood loss, the need for tracheostomy, the duration of ventilation, intensive care unit stay, and hospital stay. We also compared prevalences of acute and chronic rejection, airway complications, lung function test, and 2-year survival. RESULTS: Early allograft function in group III was significantly worse than those in groups I and II. However, there was no significant difference in any other variables of early and medium-term outcomes among the three groups. No significant correlation was detected between GIT and duration of intensive care unit stay or hospital stay. CONCLUSIONS: The limitation of acceptable GIT could be extended from the traditionally approved 4 to 5 hours, to 5 to 8 hours or even longer. PMID- 10391259 TI - Ministernotomy versus median sternotomy for aortic valve replacement: a prospective, randomized study. AB - BACKGROUND: Minimally invasive aortic valve replacement reduces surgical trauma and, supposedly, postoperative pain, blood loss, and length of stay. A prospective, randomized study was designed to prove these theoretical advantages. METHODS: Forty patients undergoing isolated, elective aortic valve replacement were randomized into two equal groups. Patients in group M underwent aortic valve replacement through a ministernotomy (reversed L or reversed C). In group S, a median sternotomy was used. The anesthetic and surgical protocol was identical for both groups. Pain was evaluated on a daily basis. Pulmonary function tests were performed preoperatively and before hospital discharge in all patients. RESULTS: There were two deaths in each group. Cross-clamp time was longer in group M: 70 +/- 19 minutes versus 51 +/- 13 minutes in group S (p = 0.005). There were no statistically significant differences between groups M and S in pump time (95 +/- 20 minutes versus 83 +/- 19 minutes), extubation time (9.9 hours in both groups), chest drainage (479 +/- 274 mL/L 24 hours versus 355 +/- 159 mL/24 hours), transfusion requirements (27% in both groups), pain evaluation (1.34 +/- 1.3 versus 2.15 +/- 1.5), length of stay (6.2 +/- 2.3 days versus 6.3 +/- 2.5 days), and cosmetic appraisal. Forced vital capacity decreased 26% from preoperative reference values in group M and 33% in group S (p = not significant). Forced expiratory volume in 1 second decreased 22% and 35%, respectively (p = not significant). CONCLUSIONS: This study has failed to prove the theoretical advantages of minimally invasive aortic valve replacement. With this technique, cross-clamp time is longer than with a median sternotomy. PMID- 10391260 TI - Spinal cord protection during aortic cross-clamping using retrograde venous perfusion. AB - BACKGROUND: Paraplegia remains a devastating complication following thoracic aortic operation. We hypothesized that retrograde perfusion of the spinal cord with a hypothermic, adenosine-enhanced solution would provide protection during periods of ischemia due to temporary aortic occlusion. METHODS: In a rabbit model, a 45-minute period of spinal cord ischemia was produced by clamping the abdominal aorta and vena cava just below the left renal vessels and at their bifurcations. Four groups (n = 8/group) were studied: control, warm saline, cold saline, and cold saline with adenosine infusion. In the experimental groups, saline or saline plus adenosine was infused into the isolated cavae throughout the ischemic period. Clamps were removed and the animals to recovered for 24 hours before blinded neurological evaluation. RESULTS: Tarlov scores (0 = paraplegia, 1 = slight movement, 2 = sits with assistance, 3 = sits alone, 4 = weak hop, 5 = normal hop) were (mean +/- standard error of the mean): control, 0.50 +/- 0.50; warm saline, 1.63 +/- 0.56; cold saline, 3.38 +/- 0.26; and cold saline plus adenosine, 4.25 +/- 0.16 (analysis of variance for all four groups, p < 0.00001). Post-hoc contrast analysis showed that cold saline plus adenosine was superior to the other three groups (p < 0.0001). CONCLUSION: Retrograde venous perfusion of the spinal cord with hypothermic saline and adenosine provides functional protection against surgical ischemia and reperfusion. PMID- 10391261 TI - Holmium:YAG laser transmyocardial revascularization relieves angina and improves functional status. AB - BACKGROUND: Transmyocardial revascularization (TMR) surgery uses laser channeling of diseased myocardium to treat ischemia and angina. Rigorous prospective randomized studies have been previously unavailable. METHODS: Forty-three patients were randomized to a medication group and 43 to a group scheduled for TMR surgery and medication. All had advanced cardiac ischemia with CCSA class 3 or 4 angina, took at least 2 cardiac medications at maximum doses, and were ineligible for angioplasty or bypass. RESULTS: Forty-two of 43 TMR group patients received surgery and were discharged after hospitalizations averaging 3.2 days. Two suffered perioperative MIs, with one death. Four others died within 12 months of surgery, 3 from cardiac events and 1 from pneumonia. Five medical group patients died from cardiac events within 12 months. Three, 6, and 12 month exams showed angina class improvement in TMR patients compared to preoperative values (3.86 +/- 0.05 vs 1.71 +/- 0.2, P < 0.0001), and to controls at 12 months (3.77 +/- 0.07 vs 1.71 +/- 0.2, P < 0.0001). Exercise tolerance improved in TMR patients over preoperative values, and was better than medication group scores after 12 months (490 +/- 17 sec. vs 294 +/- 12 sec., p = 0.0002). CONCLUSIONS: Holmium:YAG laser channeling of the myocardium improves function and reduces angina in advanced cardiac patients who lack alternative therapeutic options. PMID- 10391262 TI - Advantage of autograft and homograft valve replacement for complex aortic valve endocarditis. AB - BACKGROUND: There are advantages to using homografts and autografts as aortic valve replacements, particularly in patients with infective endocarditis. To better define these advantages, we reviewed our 13-year experience with the surgical management of infective endocarditis involving the aortic valve and root. METHODS: From 1986 through 1998, 81 adults with aortic valve endocarditis underwent valve replacement (AVR). The mean age of the 65 men and 16 women was 44 +/- 14 years. Sixty-three (78%) patients had active endocarditis at the time of operation. Non-native valve endocarditis was present in 29 (36%) patients, in 9 of whom the infection was a recurrence. Aortic valve replacements were performed with 46 homografts (homo-AVR), 25 autografts (Ross-AVR), and 10 prosthetic valves (prosth-AVR). Among Ross-AVR and homo-AVR patients, 11 required mitral valve replacement or repair (homo-Ross DVR). Follow-up was 90% complete within 2 years of the end of the study with a mean of 3.7 +/- 3.4 years. RESULTS: Early mortality was 16% (13 of 81 patients). This was 12% (3 of 25 patients) for Ross AVR, 17% (8 of 46 patients) for homo-AVR, and 20% (2 of 10 patients) for prosth AVR. Overall late mortality was 10% (7 of 68 patients) with a valve-related late mortality of 7% (5 of 68 patients). Actuarial survival at 5 years was 88% +/- 9% in Ross-AVR, 69% +/- 11% in homo-AVR, and 29% +/- 22% in prosth-AVR (p = 0.03). Endocarditis recurred in 12.5% (1 of 8 patients) with prosth-AVR and 3% (2 of 60 patients) in homo-Ross AVR. CONCLUSIONS: Valve replacement in the presence of native and prosthetic endocarditis remains a formidable challenge. Autografts and homografts are the preferred replacement aortic valves for these patients even if concomitant mitral valve replacement is required, and risk of valve-related death or recurrent endocarditis is low at medium-term follow-up. PMID- 10391263 TI - Aortic root replacement with the freestyle stentless porcine aortic root bioprosthesis. AB - BACKGROUND: Stentless porcine prosthetic valves offer several advantages over traditional valves. Among these are superior hemodynamics, laminar flow patterns, lack of need for anticoagulation and perhaps improved durability. METHODS: One hundred and twelve patients were operated on from September 17, 1992 to April 13, 1998 as part of a multi-center worldwide investigation. All patients received a total aortic root replacement. Patients were evaluated postoperatively at discharge, 3 to 6 months, and yearly by clinical exam and color flow Doppler echocardiography. RESULTS: There were 4 deaths either in the hospital or within 30 days after surgery for an operative mortality of 3.6%. No patients experienced structural valve deterioration, non-structural valve deterioration, paravalvular leak, unacceptable hemodynamic performance, or postoperative endocarditis. The linearized rates for survival and thromboembolic complications at 5 years were 82.8% and 90.5% respectively. Excellent hemodynamic function is demonstrated by very low gradients, large EOA, and an exceedingly low incidence of any aortic regurgitation. CONCLUSIONS: The Medtronic Freestyle aortic root bioprosthesis can be used safely to replace the aortic root for aortic valve and aortic root pathology. Root replacement allows optimal hemodynamic performance with no significant aortic regurgitation. Early and intermediate results are encouraging, but further follow-up is needed to determine valve durability. PMID- 10391264 TI - Cryopreserved aortic allografts for aortic root reconstruction: a single institution's experience. AB - BACKGROUND: An evaluation of early and long-term results of aortic root replacement with cryopreserved aortic allografts and echocardiographic follow-up of allograft valve function was performed. METHODS: From September 1989 through May 1998, 132 patients aged 17 to 77 years (mean, 50.8 +/- 14.8 years) underwent freestanding aortic root replacement with a cryopreserved aortic allograft. Eighty-six (65.1%) patients had New York Heart Association class III or IV functional status before operation, and 27 (20.5%) patients underwent emergency operation. Fifty-nine (44.7%) patients had undergone previous cardiac operations. The cause of aortic disease was acute endocarditis in 63 (47.7%) patients, healed endocarditis in 15 (11.3%), degenerative in 20 (15.2%), congenital in 20 (15.2%), failed prosthesis in 10 (7.6%) and rheumatic in 4 (3.0%). Follow-up was complete, with a mean of 42 months. RESULTS: There were 12 hospital deaths (9.1%; 70% confidence limits [CL], 6.6% and 11.6%); 9 of them were operated on for active endocarditis (p = 0.062). Multivariate analysis determined age older than 65 years (p = 0.012) and emergency operation (p = 0.009) as independent risk factors for hospital mortality. During follow-up, 6 (5.0%; 70% CL, 3.0% and 7.0%) patients died. Cumulative survival rate for the entire group was 81.8% +/- 5.4% at 8 years. Freedom from reoperation for structural valve failure was 100%, freedom from reoperation for any cause was 96.3% +/- 1.8% at 8 years. Freedom from endocarditis at 8 years was 97.9% +/- 1.4%. Follow-up of allograft valve function showed no or trivial aortic regurgitation in 97% of patients and absence of stenosis of the allograft in 100%. CONCLUSIONS: Aortic root replacement with cryopreserved aortic allografts can be performed with acceptable hospital mortality and long-term results. The durability of cryopreserved aortic allografts is good, and reoperation for structural valve failure is absent at 8 years. PMID- 10391265 TI - Pretreatment with a potassium-channel opener before prolonged cardiac storage: an evaluation in an experimental brain death model. AB - BACKGROUND: Pretreatment with a potassium-channel opener has been shown to improve functional recovery after long-term cardioplegic arrest. We evaluated whether pretreatment with the potassium-channel opener cromakalim is beneficial in a more clinically relevant experimental model of brain death in the rabbit. METHODS: Four groups of rabbits were studied in a 2 x 2 factorial experiment (n = 8 per group). Rabbits were subjected to a sham operation or 90 minutes of brain death induced by inflating a subdurally placed balloon. Thirty minutes before heart explantation, rabbits received either no pretreatment or an intravenous injection of cromakalim, 30 microg/kg. Hearts then received 5 hours' hypothermic storage in St. Thomas' Hospital solution and were assessed on a buffer-perfused isolated heart preparation. Hemodynamic recovery, coronary flow, and creatine kinase release were determined after 60 minutes of reperfusion. RESULTS: Systolic function and diastolic function were significantly altered in hearts explanted from brain-dead rabbits compared with hearts from rabbits having a sham operation. Cromakalim pretreatment had no significant effect on poststorage systolic or diastolic function of hearts explanted from brain-dead or sham operation rabbits. Further, cromakalim pretreatment did not affect coronary flow or overall creatine kinase release during reperfusion. CONCLUSIONS; In vivo pretreatment of brain-dead rabbits or anesthetized rabbits with an intravenous injection of cromakalim had no significant effect on functional recovery of or enzymatic release from explanted hearts after 5 hours' hypothermic storage and 60 minutes' reperfusion. These findings underscore the importance of clinically relevant experimental models. PMID- 10391266 TI - Radial artery in CABG: could the early results be comparable to internal mammary artery graft? AB - BACKGROUND: The accidental detection of patency of radial artery grafts, by Acar, which had been labeled as blocked 18 years earlier, has led to its revival as a conduit in coronary artery bypass surgery. We used radial artery as one of the grafts in 287 patients from February 1996 to June 1998. Here we present our early clinical experience and the midterm angiographic follow up of the initial 62 patients. METHODS: A no touch, atraumatic harvesting technique coupled with gentle hydrostatic and pharmacological dilatation of the radial artery graft was employed. Radial artery was used to revascularize coronary vessels with >80% proximal stenosis. Postoperatively, the patients were administered a low dose nifedipine that was continued for 6 months thereafter. The patients were followed up clinically after discharge from the hospital and angiographic evaluation of the grafted radial artery by selective injection was done at a mean interval of 16.2 +/- 5.1 months (3-24 months) postoperatively. RESULTS: There was no perioperative or late myocardial infarction or mortality. No significant complications related to the harvesting of radial artery were encountered. Angiographically, the radial artery grafts were found to be patent in 96.8% of patients (60/62). Mild distal anastomotic narrowing was seen in angiogram of one patient with good filling of the target vessel. Another patient showed diffuse spasm of radial artery graft. The patency of the pedicled left internal mammary grafts was also 98.2% (56/57). All the patients were asymptomatic. CONCLUSIONS: Radial artery seems to be an excellent alternate arterial conduit for myocardial revascularization with early and midterm patency rates equivalent to that of pedicled internal mammary artery, and it should be used more often for myocardial revascularization as an adjunct to pedicled internal mammary artery graft. PMID- 10391267 TI - Bilateral internal mammary artery grafting: midterm results of pedicled versus skeletonized conduits. AB - BACKGROUND: To increase the number of anastomoses per patient, bilateral internal mammary arteries (BIMAs) were harvested with a skeletonized approach instead of a pedicled one. METHODS: One thousand one hundred forty-six patients underwent isolated myocardial revascularization using BIMAs, 304 receiving pedicled grafts (group A, October 1991 through May 1994) and 842 receiving skeletonized conduits (group B, June 1994 through June 1998). Group B had a higher incidence of patients with diabetes (223 versus 40, p < 0.001). RESULTS: The number of BIMA anastomoses per patient was significantly higher in group B (2.4 +/- 0.3 versus 2.1 +/- 0.4, p < 0.001), as well as the number of sequential grafts (288 versus 42, p < 0.001). Twenty-three patients (2.0%) died in the first 30 days after surgery, 5 in group A (1.6%) and 18 in group B (2.1%) (not significant). Postoperative complications were similar in both groups; the incidence of sternal wound healing problems was higher as a whole and with regard to diabetic patients (4 of 40 [10%] versus 5 of 223 [2.2%], p < 0.05) in group A. Seventy-one patients in group A and 133 (15.8%) in group B underwent a postoperative angiography. Patency rate was similar, both early (100% in group A versus 98.6% in group B, not significant) and late (98.6% in group A versus 98.4% in group B, not significant). CONCLUSIONS: The use of skeletonized BIMA conduits allowed us to increase the number of BIMA anastomoses per patient with a lower rate of sternal wound complications and angiographic results similar to those obtained with pedicled BIMA conduits. PMID- 10391268 TI - Pain and quality of life after minimally invasive versus conventional cardiac surgery. AB - BACKGROUND: The aim of this study was to evaluate pain and quality of life after minimally invasive cardiac operations in comparison with conventional cardiac operations. METHODS: From October 1996 to May 1997 a total of 338 patients were interviewed daily using standard scoring systems (myocardial revascularization, n = 160; mitral valve reconstruction or replacement, n = 58; aortic valve replacement, n = 120). RESULTS: Regarding ventricular function and intensive care and hospital stay, there were no significant differences between groups. Pain decreased until the seventh postoperative day in all patients. Patients with a lateral minithoracotomy (minimally invasive revascularization and mitral valve operations) had lower pain levels from the third postoperative day onward. There were no differences in quality of life, postoperative wound healing, or stability of the bony thorax. CONCLUSIONS: In cardiac operations overall pain levels are relatively low. After minimally invasive procedures with lateral minithoracotomy, earlier mobilization is possible because of a better stability of the bony thorax, resulting in lower pain levels. PMID- 10391269 TI - Different approaches for minimally invasive closure of atrial septal defects. AB - BACKGROUND: To improve the acceptance of cosmetic results after closure of atrial septal defects, anterior or lateral thoracotomies are preferred rather than median sternotomies. Along with the availability of minimally invasive techniques, a further reduction in incision length appeared feasible while preserving thoracic stability. METHODS: Various minimally invasive approaches differing in the type of incision and mode of cannulation have been applied under conditions of normothermic ventricular fibrillation. In technique 1 (n = 5), a right parasternal mini-incision was combined with a central aortic and bicaval cannulation. Technique 2 (n = 2) was composed of an anterior submammary mini incision with femoral arterial and central bicaval cannulation. To optimize the surgical access, the transincisional cannulation of the superior vena cava was replaced by a percutaneous cervical cannulation (technique 3, n = 17). RESULTS: Effective atrial septal defect closure assessed by intraoperative echocardiography was achieved in all patients. Central neurologic complications were completely absent. Besides temporary atrial fibrillation in one case, no other cardiac complications occurred. There were no cases with complicated wound healing. CONCLUSIONS: Along with modified cannulation techniques and intraoperative echocardiography, minimally invasive techniques can be safely applied for atrial septal defect closure. Submammary incisions were highly accepted and allowed for adequate surgical exposure. PMID- 10391270 TI - Technical aspects of total revascularization in off-pump coronary bypass via sternotomy approach. AB - BACKGROUND: Cardiopulmonary bypass and cardioplegic arrest result in known physiologic inflammatory, coagulopathic, and embolic states that may result in end-organ damage. Interest in off-pump complete coronary revascularization using sternotomy exposure is therefore increasing. METHODS: Using specific surgical and anesthetic techniques, we have been able to achieve total revascularization using off-pump coronary artery bypass grafting procedures (OP-CAB) through a sternotomy approach. Exposure techniques and local stabilization are tailored to individual vessels and cardiac regions. Vascular control is achieved with silicone-elastomer loops, occluders, and shunts. Poor ventricular function, advanced age, and other comorbid conditions, in and of themselves, were not considered contraindications to OP-CAB. Cardiomegaly or situations of small, intramyocardial, or heavily calcified vessels were relative contraindications to OP-CAB. RESULTS: Of 141 sternotomy OP-CAB cases, 132 (93.6%) were completely off-pump. The mean number of OP-CAB grafts per patient in the cases that were completely off-pump was 3.3 (range, 1 to 6). The 30-day operative mortality was 0%. There were four instances of intraoperative cardiac arrest, precipitated by vascular occlusion of the right coronary artery or manipulating a cardiomegalic heart. Advanced age (> or = 80 years) or profound ventricular dysfunction (ejection fraction < or = 0.25) was present in a considerable percentage of patients (10.6% and 9.9%, respectively). CONCLUSIONS: Off-pump coronary artery bypass grafting is successful for total revascularization in large numbers of patients. Anatomic factors, including cardiomegaly and small, intramyocardial, or heavily calcified vessels are possible contraindications to OP-CAB. Patients at highest risk for undergoing cardiopulmonary bypass, including those of advanced age and having ventricular dysfunction, are precisely the ones in whom OP-CAB may be the most useful. PMID- 10391271 TI - Elevated coronary endothelin-1 but not nitric oxide in diabetics during CABG. AB - BACKGROUND: After coronary artery bypass grafting procedures, a higher incidence of morbidity and mortality has been reported in diabetic patients. We tested whether coronary artery bypass grafting in diabetics affects the endothelin-1 and nitric oxide coronary effluent profile during reperfusion. METHODS: Twenty-one consecutive patients (9 with type II diabetes mellitus, 12 non-diabetics) underwent coronary artery bypass grafting by one surgeon. The two groups did not differ in preoperative ejection fraction, Parsonnet score, number of vessels bypassed, or cross-clamp time. Each patient was treated in the same intraoperative manner with single atrial, aortic, and antegrade and retrograde cardioplegia (CPL) cannulas. Cold CPL arrest was by antegrade and retrograde infusion of modified Buckberg CPL solution. Warm CPL solution was infused before reperfusion. Coronary sinus blood samples were obtained for estimation of endothelin-1 and nitrite plus nitrate before CPL arrest and at 1 and 15 minutes after each of 2 reperfusion periods. RESULTS: In diabetics, endothelin-1 was significantly increased at all reperfusion times as compared with non-diabetics. Nitrite plus nitrate levels were significantly higher in patients with diabetes than in those without, but did not change with time in either of the groups. CONCLUSIONS: Reperfusion after CPL during coronary artery bypass grafting procedure can trigger the release of endothelin-1 in patients with diabetes mellitus. This may favor increased vascular tone or positive inotropic responses after coronary artery bypass grafting and may contribute to significant cardiovascular consequences in diabetic patients. PMID- 10391272 TI - Extended aortic replacement in acute dissection by the separated elephant trunk technique. AB - BACKGROUND: Extensive aortic replacement in acute dissection is currently not a widely accepted method of treatment. METHODS: We developed a safe method for extended aortic repair including the aortic arch in type A acute dissection, and describe here its application in 5 cases. This method was based on a modification of the elephant trunk method and several other strategies. Most of the procedures were carried out under simple hypothermic circulatory arrest. RESULTS: All patients recovered within 2 days without recurrent nerve injury. One patient suffered from unilateral upper arm palsy due to severe innominate dissection. Patients were all discharged and early postoperative computed tomography (CT) showed thrombotic obliteration around the elephant trunk. Follow-up CT after 4 to 18 months confirmed that thromboexclusion proceeded down to the distal end of the elephant graft in 1 patient and to the diaphragmatic level in 3 patients. Total obliteration was observed in the remaining 1 patient. CONCLUSIONS: This technique enables extended aortic repair in acute dissection with no increase in morbidity, and effectively promotes thromboexclusion of the dissected lumen to a wider extent than conventional methods. PMID- 10391273 TI - Determinants of cognitive change after coronary artery bypass surgery: a multifactorial problem. AB - BACKGROUND: Several studies have investigated predictors of cognitive decline after coronary artery bypass grafting (CABG), but there is little consensus as to which specific factors are predictive of poor cognitive outcomes. METHODS: We evaluated 127 patients undergoing CABG with standardized neuropsychological tests preoperatively, at 1 month and at 1 year. The outcome measure was a continuous variable reflecting change in z-scores for eight cognitive domains over time for individual patients. Univariate analyses were performed to evaluate the association between the demographic, operative, and postoperative factors and the cognitive outcome variables. Factors that were significant were included in a multiple linear regression analysis. RESULTS: Among the medical history variables, diabetes was associated with change in executive functions and psychomotor speed. Some of the operative variables were associated with short term changes, but none with the 1-year outcomes. For example, the surgeon's rating of degree of difficulty in selecting a cross-clamp site was associated with change in attention. Higher mean pump rate during the procedure was associated with improved performance on tests of language. The cognitive domains associated with medical variables were different from those associated with surgical variables, and the associations observed at 1-year were different from those seen at 1-month. CONCLUSIONS: Change in cognition after CABG is associated with both medical and surgical variables. The specifics of these associations depend on the choice of time points after surgery. This suggests that there are multiple etiologies for these changes, with nonspecific effects of anesthesia and prolonged surgery interacting with the more specific effects of the surgical procedure itself. PMID- 10391274 TI - Minimally invasive direct coronary artery bypass for redo patients. AB - BACKGROUND: The minimally invasive direct coronary artery bypass (MIDCAB) procedure, using a small anterolateral thoracotomy without cardiopulmonary bypass, has been recommended for high-risk patients because it is less traumatic than conventional coronary artery bypass grafting. For redo patients who have patent grafts and pericardial adhesions, the MIDCAB may be preferable to the conventional operation because manipulation of the graft and dissection of adhesions may be minimized. METHODS: Since November 1995, 120 patients underwent the MIDCAB procedure in our institution. Among these patients, there were 25 redo cases (20.8%). We reviewed these redo cases and studied their surgical results (mortality, morbidity, hospital stay, operation time, and postoperative inotropic support). To clarify the usefulness of this procedure, we compared the results of redo operations with those of the first-time operations. RESULTS: For redo MIDCAB, there was one operative death (4%) because of intestinal infarction. The mean hospital stay was 4.3 days and the number of patients who needed postoperative positive inotropic agents was 3 (12%). There was no significant differences between redo and first-time operation patients in mortality, morbidity, hospital stay, operation time, and postoperative inotropic support. CONCLUSIONS: Results of the MIDCAB procedure for redo patients were comparable to those for primary MIDCAB operations. PMID- 10391275 TI - Insulin improves functional and metabolic recovery of reperfused working rat heart. AB - BACKGROUND: Glucose, insulin, and potassium solution improves left ventricular function in refractory pump failure. Direct effects of insulin on the heart cannot be determined in vivo. We hypothesized that insulin has a direct positive inotropic effect on the reperfused heart. METHODS: Isolated working rat hearts were perfused with buffer containing glucose (5 mmol/L) plus oleate (1.2 mmol/L). Hearts were subjected to 15 minutes of ischemia and reperfused with or without insulin (100 microU/mL) for 40 minutes. Epinephrine (1 micromol/L) was added for the last 20 minutes. RESULTS: Hearts recovered 51.1% of preischemic cardiac power in the absence and 76.4% in the presence of insulin (p < 0.05). Whereas oleate oxidation remained unchanged, glucose uptake and oxidation increased during reperfusion with epinephrine (p < 0.01). This increase was significantly greater when hearts were reperfused in the presence of insulin (p < 0.01). Insulin also prevented an epinephrine-induced glycogen breakdown during reperfusion (p < 0.05). CONCLUSIONS: Insulin has a direct positive inotropic effect on postischemic rat heart. This effect is additive to epinephrine and occurs without delay. Increased rates of glucose oxidation and net glycogen synthesis are more protracted. PMID- 10391276 TI - Ischemic preconditioning reduces neutrophil accumulation and myocardial apoptosis. AB - BACKGROUND: This study tested the hypothesis that ischemic preconditioning (IP) inhibits myocardial apoptosis after a short period of ischemia and reperfusion. METHODS: In 9 anesthetized dogs, the left anterior descending (LAD) coronary artery was occluded for 30 min and reperfused for 3 h (control), while in 9 others, LAD occlusion was preceded by 5 min of occlusion and 5 min of reperfusion (IP). DNA from frozen myocardial tissue samples was extracted, and apoptosis were identified as "ladders" by agarose gel electrophoresis or confirmed histologically using the terminal transferase UTP nick end-labeling (TUNEL) assay. Neutrophil accumulation was detected by measuring cardiac myeloperoxidase activity. RESULTS: Thirty minutes of LAD occlusion caused a significant decrease in blood flow (colored microspheres), which was comparable between groups. In the control group, DNA ladders occurred in the area at risk (AAR) in six out nine experiments. In contrast, DNA laddering in the AAR was not observed in any of the IP group. AAR in the control group showed a greater percentage of apoptotic cells than IP (6.7 +/- 0.9% vs 1.2 +/- 0.2%; p < 0.01). Cardiac myeloperoxidase activity (U/g tissue) was significantly reduced from 0.07 +/- 0.004 in control to 0.04 +/- 0.01 in IP group (p < 0.05). CONCLUSIONS: We conclude that ischemic preconditioning attenuates apoptosis and neutrophil accumulation in the AAR in a model of nonlethal acute ischemia and reperfusion. PMID- 10391277 TI - The closed heart MAZE: a nonbypass surgical technique. AB - BACKGROUND: The MAZE-III is the surgical treatment of choice for medically refractory atrial fibrillation. Although a number of nonsurgical techniques are evolving to duplicate the transmural atrial lesions of the MAZE-III, the surgical atriotomy remains the gold standard for conduction block. It was the objective of this study to surgically create the atrial incisions of the MAZE-III without the use of cardiopulmonary bypass. METHODS: A technique was developed to create and intersect the linear incisions of the MAZE-III on 10 beating canine hearts without the use of cardiopulmonary bypass using a "tunnel" of atrial tissue. The effectiveness of the procedure was tested by atrial burst pacing. RESULTS: This technique was successfully performed on 10 mongrel dogs without operative mortality. Preoperatively, sustained atrial fibrillation (>30 seconds) was induced in all animals. Postoperatively, all the animals remained in sinus rhythm even after burst pacing. CONCLUSIONS: In an experimental canine model, the MAZE III can be performed on beating hearts without the assistance of cardiopulmonary bypass using a "tunnel" technique. This technique allows for the immediate assessment of electrophysiologic and mechanical function after the MAZE-III, or any other type of procedure using the "maze principle" and may find future application in the clinical arena. PMID- 10391278 TI - Early experience with partial left ventriculectomy in the Asia-Pacific region. AB - BACKGROUND: We report our early experience with partial left ventriculectomy done by a group of cardiac surgeons in the Asia-Pacific region. METHODS: Partial left ventriculectomy was done in 48 patients (mean age, 43 years) with advanced symptomatic cardiomyopathy. The origin of cardiomyopathy was idiopathic in 30 patients, valvular in 10, ischemic in 3, peripartum in 3, sarcoidosis in 1, and viral myocarditis in 1. Procedures performed on the mitral valve were repair with Alfieri method in 8 patients, ring annuloplasty in 2, and replacement in 25. RESULTS: Seventy-seven percent of patients required myocardial support for weaning from cardiopulmonary bypass, and the overall in-hospital mortality rate was 27%. Mean follow up was 6.5 months (range, 1 to 18 months), and patient survival at 1, 3, and 6 months after discharge was 91%, 88%, and 84%, respectively. Sixty-five percent of survivors with idiopathic and valvular disease achieved significant and sustained improvement in ventricular contractility and symptoms, but there were no clear symptomatic benefits from partial left ventriculectomy in patients with cardiomyopathy from other causes. Most cases of late recurrence of heart failure symptoms (90%) appeared to be related to the development of progressive mitral incompetence. CONCLUSIONS: After partial left ventriculectomy left ventricular function improved in patients with idiopathic and valve related cardiomyopathies. Late deterioration was related to the development of significant mitral valve incompetence postoperatively, hence definitive mitral valve repair or replacement at the time of the partial left ventriculectomy procedure is advised. PMID- 10391279 TI - Transmyocardial laser revascularization: experimental studies in healthy porcine myocardium. AB - BACKGROUND: Clinical studies have demonstrated a significant reduction of cardiac index shortly after transmyocardial laser revascularization in patients with low ejection fraction. We analyzed the influence of transmyocardial laser revascularization on healthy myocardium in pigs. METHODS: Carbon dioxide channels were created in 20 pigs which were observed for 6 hours. Ten pigs received one laser channel and ten pigs two laser channels per cm2 in the left anterior descending artery region. Seven pigs served as controls. Perfusion (microspheres), function, histochemical, and histologic assessments were subsequently performed. RESULTS: A significant deterioration of left ventricular stroke work index was observed shortly after transmyocardial laser revascularization in both laser groups (p < 0.05). After 6 hours the left ventricular stroke work index did not increase and showed significantly reduced values at rest (p < 0.05) and during stress in the laser groups (p < 0.01). Normal regional perfusion, small ischemic and necrotic areas, open laser channels in the left anterior descending artery region and significantly increased myocardial water content were observed in the laser groups (p < 0.01). CONCLUSIONS: Carbon dioxide laser channels significantly decrease global heart function shortly after transmyocardial laser revascularization in healthy porcine myocardium. This myocardial tissue showed no recovery 6 hours postoperatively. PMID- 10391280 TI - Improved perfusion and contractile reserve after transmyocardial laser revascularization in a model of hibernating myocardium. AB - BACKGROUND: Transmyocardial laser revascularization (TMR) has been demonstrated effective for relieving angina, although prior studies have yielded inconsistent results regarding postoperative myocardial perfusion and function. This study evaluated long-term changes in myocardial perfusion and contractile reserve after TMR in a model of hibernating myocardium. METHODS: Miniswine had subtotal left circumflex coronary artery occlusion to reduce resting blood flow to 10% of baseline. After 2 weeks in the low-flow state, positron emission tomography and dobutamine stress echocardiography were performed to document ischemic, viable (hibernating) myocardium in the left circumflex distribution. Animals then had sham redo thoracotomy (n = 4) or TMR (n = 6). Six months later the positron emission tomography and dobutamine stress echocardiography studies were repeated. RESULTS: Myocardial blood flow in the left circumflex distribution as measured by positron emission tomography was significantly reduced in all animals after 2 weeks in the low-flow state. In animals that had TMR, there was significant improvement in myocardial blood flow to the lased regions 6 months postoperatively. No significant change in myocardial blood flow was seen in sham animals at 6 months. Dobutamine stress echocardiography after 2 weeks of low-flow demonstrated severe hypocontractility at rest in the left circumflex region of all animals, with a biphasic response to dobutamine consistent with hibernating myocardium. In animals that had TMR, there was a trend toward improved resting function and significantly improved regional stress function in the lased segments 6 months postoperatively, consistent with a reduction in ischemia. Global left ventricular wall motion at peak stress improved significantly as well. There was no change in wall motion 6 months postoperatively in sham operated animals. CONCLUSIONS: This study found improvements in myocardial perfusion and regional and global contractile reserve 6 months after TMR in a porcine model of hibernating myocardium. This improved perfusion and function likely accounts for the clinical benefits of the procedure. PMID- 10391281 TI - Release of S100B during coronary artery bypass grafting is reduced by off-pump surgery. AB - BACKGROUND: S100B, a plasma marker of brain injury, was compared after coronary artery bypass grafting with and without cardiopulmonary bypass (CPB). METHODS: Fourteen patients with off-pump operations and 18 patients with CPB were compared. Seven patients in the off-pump group underwent a minithoracotomy and received only an arterial graft, whereas 7 patients underwent sternotomy and received both an arterial and one or two vein grafts. S100B was measured in arterial plasma using an immunoassay with enhanced sensitivity. RESULTS: S100B before the operation was 0.03 microg/L. At wound closure, S100B in patients of the off-pump and CPB groups reached a maximum level of 0.22 +/- 0.07 and 2.4 +/- 1.5 microg/L, respectively (p < 0.001). No strokes occurred. Patients without CPB receiving arterial and vein grafts released slightly more S100B (p < 0.05) than patients with only arterial grafting. In patients undergoing CPB, S100B increased slightly before aortic cannulation (p < 0.001), to the same level as the maximum reached for the non-CPB group. CONCLUSIONS: Coronary artery bypass grafting with CPB caused a 10-fold greater increase in S100B than off-pump grafting. S100B release after off-pump sternotomy with vein grafting was slightly greater than in arterial grafting through a minithoracotomy. PMID- 10391283 TI - Adenosine A3 pretreatment before cardioplegic arrest attenuates postischemic cardiac dysfunction. AB - BACKGROUND: The cardioprotective effects of the adenosine A3 receptor in a cardioplegia model have not been described. We tested the hypothesis that infusion of the A3 receptor agonist, Cl-IB-MECA (100 nM), as a pretreatment (PTx) and/or as a cardioplegic (CP) additive reduces postischemic myocardial injury. METHODS: Isolated perfused rat hearts underwent 30 minutes of normothermic ischemia, 60 minutes of intermittent hypothermic cardioplegia (10 degrees C), followed by 2 hours of reperfusion. Hearts were divided into four groups: (1) no pretreatment (PTx) and unsupplemented cardioplegia (CP) (control), (2) Cl-IB-MECA PTx and unsupplemented CP (A3-PTx), (3) no PTx and Cl-IB-MECA CP (A3-CP), or (4) Cl-IB-MECA PTx and Cl-IB-MECA CP (A3-[PTx+CP]). RESULTS: Coronary flow was not increased after A3 pretreatment when compared to baseline values. After 2 hours of reperfusion, left ventricular developed pressure in control and A3-CP groups was depressed to 43% +/- 3% and 47% +/- 2% of baseline; while A3-PTx and A3 [PTx+CP] significantly increased left ventricular developed pressure (65% +/- 3% and 61% +/- 5%) from baseline relative to control and A3-CP. Effluent creatine kinase activity was significantly decreased by A3-PTx (1520 +/- 32 IU/L), A3 [PTx+CP] (1481 +/- 41 IU/L) from control (1734 +/- 54 IU/L) and A3-CP (1750 +/- 43 IU/L). Myocardial edema (% tissue water) was significantly less in A3-PTx (78 +/- 0.6%) and A3-[PTx+CP] (76% +/- 2%) compared with control (85% +/- 0.4%) and A3-CP (83% +/- 2%). CONCLUSIONS: Adenosine A3 receptor stimulation as a pretreatment attenuates postischemic cardiodynamic dysfunction and creatine kinase release but has no cardioprotection as an adjunct to cold cardioplegia. PMID- 10391282 TI - Gene transfection of hepatocyte growth factor attenuates reperfusion injury in the heart. AB - BACKGROUND: Hepatocyte growth factor (HGF), a ligand for the c-Met receptor tyrosine kinase, plays a role as organotrophic factor for regeneration of various organs. HGF has an angiogenic activity and exhibits a potent antiapoptotic activity in several types of cells. Although HGF and the c-Met/HGF receptor are expressed in the heart, the role of HGF in the heart has remained unknown. METHODS: After we analyzed changes in expression of endogenous HGF and c-Met mRNA levels in the rat left ventricle after myocardial infarction, the human HGF gene in hemagglutinating virus of Japan (HVJ)-liposome was transfected into the normal whole rat heart. Three days after transfection, the heart was subjected to global warm ischemia and subsequent reperfusion, followed by assessment of its cardiac functions. RESULTS: Both HGF and c-Met/HGF receptor mRNAs were expressed in adult rat heart, and c-Met/HGF receptor mRNA was upregulated in response to myocardial infarction. HGF-transfected heart showed significant increase of human HGF protein level in the heart. Cardiac functions in terms of the left ventricular developed pressure, maximum dp/dt, and pressure rate product in hearts with HGF gene transfection were significantly superior to those in control hearts. In addition, leakage of creatine phosphokinase in the coronary artery effluent in hearts with HGF gene transfection was significantly lower than that in control hearts. CONCLUSIONS: These data indicated that both HGF and c-Met/HGF receptor mRNAs were upregulated in response to myocardial ischemic injury, and that HGF is likely to have a cytoprotective effect on cardiac tissue, presumably through the c-Met/HGF receptor. PMID- 10391284 TI - Pulmonary artery sling: results with median sternotomy, cardiopulmonary bypass, and reimplantation. AB - BACKGROUND: The classic surgical approach to pulmonary artery (PA) sling has been through a left thoracotomy with division of the left PA and reimplantation into the main PA anterior to the trachea. Another approach is anterior left PA translocation with distal tracheal resection. Since 1985, we have repaired PA sling with a median sternotomy approach, cardiopulmonary bypass, and division and reimplantation of the left PA into the main PA with simultaneous repair of associated tracheal stenosis. The purpose of this review is to determine the outcome of that strategy. METHODS: From 1985 to 1998, 16 infants had surgical treatment of PA sling, 14 had left PA division and reimplantation into the MPA, 2 patients had repair using the translocation technique. Mean age at repair was 6.9 months, median age was 4 months. All infants, except 1 with an absent right lung, were operated on at the time of diagnosis. All had rigid bronchoscopy, which revealed associated complete tracheal rings in 12 patients. Seven patients had tracheal repair with pericardial tracheoplasty, 4 had repair using a tracheal autograft technique, and 2 had a distal tracheal resection (one for tracheomalacia). Of the 2 patients having the translocation technique, 1 had a severely hypoplastic right lung and the other had complete absence of the right lung. RESULTS: There has been no operative mortality. Hospital stay ranged from 5 to 188 days (mean 36 +/- 42 days). There was 1 late death 7 months postoperatively from respiratory complications of pericardial tracheoplasty. All left pulmonary arteries are patent and blood flow to the left lung by nuclear scan (n = 10) ranges from 24% to 46% (mean 35% +/- 9%). CONCLUSION: The strategy of median sternotomy, cardiopulmonary bypass, and left PA division and reimplantation into the main PA with simultaneous tracheal repair has resulted in a low operative mortality and excellent patency of the left pulmonary artery. Results with repair of the commonly associated complete tracheal rings has recently improved with the use of the free tracheal autograft technique. PMID- 10391285 TI - The modified Fontan procedure: morphometry and surgical implications. AB - BACKGROUND: The modified Fontan procedure for patients with only one well-formed ventricle is now widely regarded as palliative, not curative. METHODS: To improve the surgical management and postoperative follow-up of such patients, a morphometric study of 33 postmortem cases was done. RESULTS: The three main causes of death were congestive heart failure (82%), arrhythmias (12%), and central nervous system dysfunction (6%). The cross-sectional area of the Fontan anastomosis (FA) relative to the systemic venous area (SVA) and relative to the body surface area (BSA) revealed that the Fontan pathway was often obstructive. The mean FA/SVA index was 73% less than normal: 0.54 +/- 0.22, range 0.13 to 0.98. The mean FA/BSA index was 70% less than normal: 143.52 +/- 50.01 mm2/M2, range 55.09 to 261.67 mm2/M2. CONCLUSIONS: The main surgical challenge is to minimize or eliminate prepulmonary stenosis. Although significant postoperative obstruction was often not evident hemodynamically because of small or absent gradients, the presence of important obstruction of the Fontan pathway was clearly revealed by morphometry. PMID- 10391286 TI - "Swiss cheese" septal defects: surgical closure using a single patch with intermediate fixings. AB - BACKGROUND: Residual ventricular septal defects and ventricular and septal dysfunctions are surgical drawbacks of "Swiss cheese" defects. We developed a technique that uses a single patch with intermediate fixings to cover the right side of the septum without producing a septal bulging, through a right atriotomy. METHODS: Since April 1993, 5 children with "Swiss cheese" defects have been operated on using this procedure (mean age, 17 +/- 12 months). Three patients had associated lesions including tetralogy of Fallot, Taussig Bing heart, and mitral stenosis. RESULTS: There have been no early or late deaths. The mean follow-up time is 29 +/- 18 months. All patients are asymptomatic. Echocardiography revealed either an intact septum (n = 4) or insignificant color jets at the apical portion of the septum (n = 1). The septal wall motion was preserved in 4 children and was hypokinetic in the fifth child. CONCLUSIONS: This technique can be an additional tool to provide a secure closure of "Swiss cheese" defects even in the presence of associated cardiac lesions. Long-term consequences of this procedure on septal wall motion remain to be determined. PMID- 10391287 TI - Addition of a small curvature reduces power losses across total cavopulmonary connections. AB - BACKGROUND: In the Fontan circulation the vis a tergo for lung perfusion is limited. The hypothesis of this in vitro study was that energy dissipation at the common cavopulmonary connection can be reduced by the addition of caval curvature. METHODS: Two Perspex models were analyzed, the commonly used crosslike cavopulmonary connection (model 1) and a modified curved configuration (model 2). Pressures and flows across the connections were measured simultaneously at various caval and pulmonary artery flow splits and resistances. Mixing of inferior and superior caval fluid was evaluated. RESULTS: Caval pressure oscillations occurred in model 1 only. Curvature reduced power losses in all settings significantly (alpha = 0.05), most successfully at adult caval flow ratios and at high flow rates. At equal pulmonary resistances pulmonary flow was balanced in both models. The inferior caval fluid is preferably directed to the right lung in model 2 predominantly for caval flow conditions in younger patients. CONCLUSIONS: Our data show that the modified curved cavopulmonary connection is hydrodynamically advantageous but might impair caval fluid mixing in younger children. PMID- 10391288 TI - Delayed impairment of cerebral oxygenation after deep hypothermic circulatory arrest in children. AB - BACKGROUND: Clinical studies of deep hypothermic circulatory arrest (DHCA) have focused only on the immediate postoperative period. However, experimental findings suggest impairment of cerebral oxygenation at 2 to 8 hours after reperfusion. METHODS: In 10 children who had DHCA for heart operations, transcerebral differences of hemoglobin oxygen saturation and plasma hypoxanthine, xanthine, and lactoferrin concentrations were measured in concurrently obtained cerebral venous, arterial, and mixed venous samples up to 10 hours postoperatively. RESULTS: Compared with preoperative levels (57% +/- 7%), cerebral venous oxygen saturation was not significantly reduced until 2 hours (44% +/- 6%) and 6 hours (42% +/- 5%) after DHCA (p < 0.05). A statistically significant transcerebral (ie, cerebral vein versus artery) concentration difference of hypoxanthine was observed at 30 minutes (3.6 +/- 0.9 micromol/L), 1 hour (3.4 +/- 1.1 micromol/L), and 2 hours (3.1 +/- 0.8 micromol/L) after DHCA but not preoperatively (0.4 +/- 0.2 micromol/L). A transcerebral concentration difference of lactoferrin occurred 30 minutes after DHCA (196 +/- 70 microg/mL) but not preoperatively (16 +/- 20 microg/mL). CONCLUSIONS: Cerebral venous oxygen saturation of hemoglobin decreased as late as 2 to 6 hours after DHCA, in association with impaired cerebral energy status. Neutrophil activation in the cerebral circulation occurred 30 minutes after reperfusion. PMID- 10391289 TI - Novel techniques of bidirectional Glenn shunt without cardiopulmonary bypass. AB - BACKGROUND: We report novel techniques of performing bidirectional Glenn shunt (BDG) without cardiopulmonary bypass (CPB). METHODS: Five cases of single ventricle and pulmonary stenosis (PS) complex were taken up for BDG without CPB. The criteria for case selection were an unrestrictive atrial septal defect (ASD), no atrioventricular (AV) valve regurgitation, and no other intracardiac defects requiring correction. A temporary shunt was established between the superior vena cava (SVC) and contralateral branch pulmonary artery (PA) for venous drainage during SVC clamping for BDG anastomosis in four cases. In case 5, a shunt was put between the SVC and right atrium (RA) for venous drainage, and modified Blalock Taussig shunt and patent ductus arteriosus (PDA) were left open until the completion of the BDG. RESULTS: Central venous pressure (CVP) increased to a mean of 22.4 mm Hg during SVC clamping, with improvement of oxygen (O2) saturation from 62.4% to 82.4%. After Glenn shunt, CVP and O2 saturation maintained at 13.2 mm Hg and 87.4%, respectively. Postoperatively, there were no neurological abnormalities and no hospital mortality. CONCLUSIONS: Our technique provides an excellent venous drainage with improvement of O2 saturation during SVC clamping. It avoids problems related to CPB and economy. It is easily reproducible, with excellent results in a selected group of patients without compromising the completeness of repair. PMID- 10391290 TI - Immediate vein graft thrombectomy for acute occlusion after coronary artery bypass grafting. AB - A 76-year-old man underwent coronary bypass grafting 3 days after exposure to heparin. Immediately after chest closure, he developed acute graft thrombosis and cardiac arrest in the setting of thrombocytopenia. Immediate graft thrombectomies were performed. Postoperative tests for heparin-induced thrombocytopenia and thrombosis (HITT) were positive. This case represents a dramatic example of HITT after coronary revascularization. PMID- 10391291 TI - An unusual case of hypoxia from benzocaine-induced methemoglobinemia. AB - Hypoxemia during bronchoscopy occurs frequently. It can usually be managed by supplemental oxygen and bronchodilators or, in some cases, occasionally stopping the procedure. Benzocaine spray is commonly used as a topical anesthetic agent during bronchoscopy. However, it has been associated with the development of methemoglobinemia. The following is a case report of hypoxia during bronchoscopy from benzocaine-induced methemoglobinemia and its management. PMID- 10391292 TI - Ventricular thrombus and subarachnoid bleeding during support with ventricular assist devices. AB - We report the case of a 23-year-old man with acute aortic valve insufficiency caused by endocarditis, who after emergency aortic valve replacement developed biventricular heart failure. The heart failure was treated with temporary assist devices. Subarachnoid bleeding and thrombus obstruction of the left ventricular outflow tract was detected. The postoperative course is presented with special emphasis on management of subarachnoid bleeding and the simultaneous use of anticoagulation necessary for ventricular assist devices. PMID- 10391293 TI - Successful treatment of a ruptured mycotic coronary artery aneurysm. AB - Documented mycotic aneurysms of the coronary arteries are unusual, and antemortem identification of such an aneurysm is rare. We present the case of a patient who had successful management of a ruptured mycotic aneurysm of a coronary artery. PMID- 10391294 TI - Fibrous ball: a new manifestation of chronic defibrillator and pacemaker infection. AB - Two patients with unusual manifestation of long-term infection of implantable cardioverter defibrillator and pacemaker were examined. Complete explanation of the defibrillator and pacemaker was done in both patients. New devices were subsequently implanted. PMID- 10391295 TI - Coronary revascularization with arterial conduits collateral to the lower limb. AB - A 62-year-old man with left main coronary artery disease had coronary artery bypass grafting. Angiography disclosed total occlusion of the left common iliac artery. The left internal thoracic artery and the left inferior epigastric artery were well developed as collateral pathways to the left external iliac artery. With concomitant femoro-femoral crossover bypass, these two large arterial conduits were harvested and grafted to the coronary artery. PMID- 10391296 TI - Successful thrombolysis for massive pulmonary embolism after pulmonary resection. AB - We report the successful use of thrombolysis for acute massive pulmonary embolism 2 days after right lower lobectomy for bronchial adenocarcinoma. Pulmonary angiography revealed extensive clot unsuitable for surgical embolectomy. A bolus infusion of recombinant tissue plasminogen activator produced an immediate improvement in the patient's hemodynamic state. There was substantial blood loss requiring the transfusion of 21 units of blood over the postoperative period. The patient made a successful recovery and remained well at 1 year. PMID- 10391297 TI - Pulmonary thromboembolectomy of donor lungs prior to lung transplantation. AB - Successful bilateral single-lung transplantation was performed after pulmonary thromboembolectomy of the donor lungs. The donor lungs were not thought to contain large amounts of pulmonary thromboemboli because they satisfied all the donor selection criteria. This case reinforces the need of not only meticulous inspection of the donor lungs prior to implantation but also the productive use of available donor organs. PMID- 10391298 TI - Mediastinal bronchogenic cyst manifesting as a catastrophic myocardial infarction. AB - Congenital bronchogenic cysts of the lung and mediastinum develop from the ventral foregut during embryogenesis. These cysts are often incidental radiologic findings in adults, but patients can be seen with symptoms of chest pain, cough, dyspnea, or any combination of these. Acute presentations are unusual and have rarely been reported. We present the unique case of a 36-year-old man seen with an acute coronary syndrome and sudden hemodynamic collapse. The patient sustained a massive and ultimately fatal myocardial infarction, compression of the left main coronary artery by a bronchogenic cyst was demonstrated at postmortem examination. If detected, bronchogenic cysts should be surgically excised to limit associated morbidity and mortality. PMID- 10391299 TI - Cardiac echinococcosis causing coronary artery disease. AB - We report a case of cardiac echinococcosis in a patient who had sustained an acute myocardial infarction 1 month previously. The coronary angiographic study revealed an isolated complete intrinsic obstruction of the left anterior descending coronary artery in the area of the cyst. The cyst was surgically removed. Coronary artery bypass grafting was not performed because of distal myocardial scarring. Coronary arteriography should routinely be performed in all patients with cardiac echinococcosis. PMID- 10391300 TI - Cardiac autotransplant for surgical treatment of a malignant neoplasm. AB - Because of their anatomic location, cardiac sarcomas often interfere with cardiac function. Excision is considered to palliate the cardiac defect, but complete excision is often difficult owing to access, particularly in left atrial tumors. Incomplete resection results in tumor recurrence. To achieve complete resection of a large left atrial sarcoma, we used the technique of cardiac explantation, extracorporeal resection of the tumor with cardiac reconstruction, and cardiac autotransplantation. PMID- 10391301 TI - Second primary Barrett's adenocarcinoma after 19 years. AB - Because long survival after resection of esophageal carcinoma is uncommon, second esophageal cancers are rare. We report the case of a patient in whom adenocarcinoma developed within residual Barrett's esophagus 19 years after esophagectomy for stage IIb Barrett's adenocarcinoma. Implications relative to the type of operation and adequacy of resection are discussed. Long-term survival after Barrett's adenocarcinoma may occur more often if surveillance protocols achieve their aim. Questions concerning the management of such patients are identified. PMID- 10391302 TI - Repair of an ascending aorta pseudoaneurysm by way of superior ministernotomy. AB - A wide ascending aorta pseudoaneurysm occurring 10 years after uncomplicated aortic valve replacement was successfully repaired using a superior ministernotomy and femoral-femoral cannulation. In this setting, a limited sternal incision minimized the risk of pseudoaneurysm rupture during dissection and allowed safe isolation of the target cardiac structures. PMID- 10391303 TI - Late vasospasm of the inferior epigastric artery graft. AB - We report a case of vasospasm of a free inferior epigastric artery graft at 5 year angiographic follow-up after coronary artery bypass grafting. Although the cause of spasm was not clear, the graft was viable and had a vasoconstrictor profile similar to a gastroepiploic artery graft at long-term follow-up. PMID- 10391304 TI - Repair of left ventricular rupture after mitral valve replacement: use of a Teflon patch and glue. AB - Rupture of the left ventricular wall is an infrequent but lethal complication after mitral valve replacement. We present the case of a patient in whom such a rupture was successfully repaired in the intensive care unit with a patch of Teflon felt stuck in place with glue. PMID- 10391305 TI - Tricuspid valve myxoma in a pediatric patient: 10-year follow-up after resection. AB - Cardiac myxomas are rarely encountered in pediatric patients. Tricuspid valve involvement in these cases is even more exceptional. We report the case of a 5 year-old girl operated on successfully 10 years ago for a tricuspid valve myxoma who continues to be asymptomatic and had an event free outcome. PMID- 10391306 TI - Green aortic valve: alcaptonuria (ochronosis) with severe aortic stenosis. PMID- 10391307 TI - Reversed "C" ministernotomy for aortic valve replacement. AB - The technique of aortic valve replacement through a reversed "C" sternotomy incision is described. The sternal incision extends between the second and the fifth intercostal space and provides excellent exposure of the ascending aorta, the aortic root, and the right atrial appendage. The procedure can be performed with standard cannulation for cardiopulmonary bypass and conventional surgical instruments. PMID- 10391308 TI - Video-assisted approach for transxiphoid bilateral lung metastasectomy. AB - Radical resection has proved to be the most effective treatment of lung metastases, and manual palpation is considered the most accurate method for detection of occult metastases. To allow bilateral manual palpation during video assisted metastasectomy, we developed a transxiphoid approach without sternotomy. Twenty-one lesions were successfully resected in 6 patients without mortality or morbidity. This approach allows easy manual palpation of the lungs and facilitates bilateral video-assisted metastasectomy. PMID- 10391309 TI - A simplified method of stabilization and hemostasis for minimally invasive coronary artery bypass. AB - We describe a simple method to achieve both hemostasis and stabilization of the left anterior descending coronary artery during minimally invasive coronary artery bypass grafting. This technique allows the surgeon to perform a precise anastomosis of the left internal mammary artery to the target vessel on a beating heart. PMID- 10391310 TI - The incisional pulmonary artery band. AB - Occasionally early definitive repair of congenital heart disease carries prohibitive mortality, and interval pulmonary artery banding is necessary to protect the pulmonary arterial bed and improve systemic perfusion or prepare a systemic left ventricle for a later arterial switch operation. We describe our technique for effectively banding the pulmonary artery. PMID- 10391311 TI - Retrograde cerebral perfusion for aortic operations through left thoractomy. AB - Retrograde cerebral perfusion during deep hypothermic circulatory arrest is a technique used largely during operations on the ascending aorta, aortic arch, or both through a median sternotomy. This method is not frequently used for operations performed through a left thoracotomy because of problematic access to the right side of the heart. We propose a technique allowing retrograde cerebral perfusion through a left thoracotomy in a quick, simple, and efficient manner. PMID- 10391312 TI - A new method of myocardial revascularization with the radial artery. AB - We present a new method of myocardial revascularization. The radial artery is used in combination with the left internal mammary artery, thereby providing three distal end-to-side anastomoses to the left anterior descending coronary artery and other sites as determined by the coronary artery lesions. Arterial conduits form an anastomotic network between the left internal mammary artery and the radial artery in a horseshoe pattern. Three coronary arteries are revascularized by two arterial conduits in the left coronary system. PMID- 10391313 TI - Valve replacement for appetite suppressant-induced valvular heart disease. AB - Valvular heart disease associated with the use of appetite-suppressant medication is a recently described clinical entity. Although the mechanism of valvular injury remains elusive pathologically, the valvular abnormalities resemble those observed in carcinoid syndrome. The incidence of clinically evident valvular heart disease is low with short-term (less than 3 months) exposure to appetite suppressant drugs. Prolonged exposure to higher doses in addition to combination drug therapy confers an excess risk for valvular pathologic changes. We report the case of a patient with severe mitral regurgitation who had short-term exposure (3 weeks) to the combination of fenfluramine (20 mg) and phenteramine (15 mg). PMID- 10391314 TI - As originally published in 1992: Chest wall stabilization with synthetic reabsorbable material. Updated in 1999. PMID- 10391315 TI - Use of radial artery as coronary bypass graft in myocardial revascularization. PMID- 10391316 TI - Surgical management of flail chest. PMID- 10391317 TI - An unexpected benefit of the radial artery. PMID- 10391318 TI - Current review of blood activation: actually a call for papers. PMID- 10391319 TI - Treatment modalities for thoracic empyema: the right indication for the right disease. PMID- 10391320 TI - Surgical treatment of the dilated ascending aorta: when and how? AB - BACKGROUND: The aorta is considered pathologically dilated if the diameters of the ascending aorta and the aortic root exceed the norms for a given age and body size. A 50% increase over the normal diameter is considered aneurysmal dilatation. Such dilatation of the ascending aorta frequently leads to significant aortic valvular insufficiency, even in the presence of an otherwise normal valve. The dilated or aneurysmal ascending aorta is at risk for spontaneous rupture or dissection. The magnitude of this risk is closely related to the size of the aorta and the underlying pathology of the aortic wall. The occurrence of rupture or dissection adversely alters natural history and survival even after successful emergency surgical treatment. METHODS: In recommending elective surgery for the dilated ascending aorta, the patient's age, the relative size of the aorta, the structure and function of the aortic valve, and the pathology of the aortic wall have to be considered. The indications for replacement of the ascending aorta in patients with Marfan's syndrome, acute dissection, intramural hematoma, and endocarditis with annular destruction are supported by solid clinical information. Surgical guidelines for intervening in degenerative dilatation of the ascending aorta, however, especially when its discovery is incidental to other cardiac operations, remain mostly empiric because of lack of natural history studies. The association of a bicuspid aortic valve with ascending aortic dilatation requires special attention. RESULTS: There are a number of current techniques for surgical restoration of the functional and anatomical integrity of the aortic root. The choice of procedure is influenced by careful consideration of multiple factors, such as the patient's age and anticipated survival time; underlying aortic pathology; anatomical considerations related to the aortic valve leaflets, annulus, sinuses, and the sino-tubular ridge; the condition of the distal aorta; the likelihood of future distal operation; the risk of anticoagulation; and, of course, the surgeon's experience with the technique. Currently, elective root replacement with an appropriately chosen technique should not carry an operative risk much higher than that of routine aortic valve replacement. Composite replacement of the aortic valve and the ascending aorta, as originally described by Bentall, DeBono and Edwards (classic Bentall), or modified by Kouchoukos (button Bentall), remains the most versatile and widely applied method. Since 1989, the button modification of the Bentall procedure has been used in 250 patients at Mount Sinai Medical Center, with a hospital mortality of 4% and excellent long-term survival. In this group, age was the only predictor of operative risk (age > 60 years, mortality 7.3% [9/124] compared with age < 60, mortality 0.8% [1/126], p = 0.02). CONCLUSIONS: This modification of the Bentall procedure has set a standard for evaluating the more recently introduced methods of aortic root repair. PMID- 10391321 TI - Aortic valve sparing operations: an update. AB - BACKGROUND: Aortic valve sparing operations in patients with ascending aorta and/or aortic root aneurysms have been performed for a decade in our institution. Initially only patients with normal aortic valve leaflets had these operations, but more recently we utilized them in patients with prolapse of a single leaflet and in those with a bicuspid aortic valve. This article is an update on the clinical results of these operations. METHODS: From May 1988 to December 1997, 126 patients with ascending aorta and/or aortic root aneurysms and aortic insufficiency underwent replacement of the ascending aorta with reconstruction of the aortic root and preservation of the native aortic valve. There were 85 men and 41 women, with a mean age of 54 years (range, 14 to 84). Thirty-two patients had the Marfan syndrome; 17 patients had acute and 10 had chronic type A aortic dissection; 23 had a transverse arch aneurysm; 26 had coronary artery disease, and 8 had mitral regurgitation. The aortic valve sparing operation consisted of simple adjustment of the sinotubular junction in 33 patients, adjustment of the sinotubular junction and replacement of one or more aortic sinuses in 60, and reimplantation of the aortic valve in a tubular Dacron (C.R. Bard, Haverhill, PA) graft in 33. Fifteen patients also had repair of aortic leaflet prolapse. Only 4 patients had a bicuspid aortic valve. RESULTS: There were 3 operative deaths due to cardiac failure. Patients were followed from 2 to 117 months, with a mean of 31. There were 11 late deaths: 7 cardiovascular and 4 from unrelated causes. The actuarial survival was 72 +/- 8% at 7 years. Two patients required aortic valve replacement; the freedom from aortic valve replacement was 97 +/- 2% at 7 years. Doppler echocardiography revealed absent, trivial or mild aortic insufficiency in most patients; only 9 patients had moderate aortic insufficiency. CONCLUSIONS: Aortic valve sparing operations are feasible in most patients with ascending aorta and/or aortic root aneurysms who have normal or near normal aortic leaflets. The functional results of the repaired aortic valve are excellent, and the repair appears to be durable. PMID- 10391322 TI - Ross procedure for ascending aortic replacement. AB - BACKGROUND: Patients with aortic valve disease and aneurysm or dilatation of the ascending aorta require both aortic valve replacement and treatment of their ascending aortic disease. In children and young adults, the Ross operation is preferred when the aortic valve requires replacement, but the efficacy of extending this operation to include replacement of the ascending aorta or reduction of the dilated aorta has not been tested. METHODS: We reviewed the medical records of 18 (5.9%) patients with aortic valve disease and an ascending aortic aneurysm and 26 (8.5%) patients with dilation of the ascending aorta, subgroups of 307 patients who had a Ross operation between August 1986 and February 1998. We examined operative and midterm results, including recent echocardiographic assessment of autograft valve function and ability of the autograft root and ascending aortic repair or replacement to maintain normal structural integrity. RESULTS: There was one operative death (2%) related to a perioperative stroke. Forty-two of 43 survivors have normal autograft valve function, with trace to mild autograft valve insufficiency, and one patient has moderate insufficiency at the most recent echocardiographic evaluation. None of the patients has dilatation of the autograft root or of the replaced or reduced ascending aorta. CONCLUSIONS: Early results with extension of the Ross operation to include replacement of an ascending aortic aneurysm or vertical aortoplasty for reduction of a dilated ascending aorta are excellent, with autograft valve function equal to that seen in similar patients without ascending aortic disease. PMID- 10391323 TI - Aortic allografts and pulmonary autografts for replacement of the aortic valve and aortic root. AB - BACKGROUND: Extensive experience has accumulated with the use of aortic and pulmonary autografts for replacement of the aortic valve and the aortic root. Three general techniques for insertion have been used: subcoronary (free-hand) valve implantation, mini- or inclusion-root implantation, and aortic root replacement. Thirty-day mortality for elective operations with all of these techniques has not exceeded 5%. Thromboembolic episodes have been rare, and endocarditis has occurred infrequently. Early hemodynamic performance has been excellent, without significant gradients or valve regurgitation in the majority of patients. METHODS AND RESULTS: Progressive aortic regurgitation has been observed with continued follow-up, and is the most important complication of both types of valves. Leaflet failure and technical problems are the major causes of reoperation for patients receiving aortic allografts. There is some evidence to suggest that the prevalence of these complications is lower with the root replacement technique than with the intraaortic implantation methods. CONCLUSIONS: Reoperation for regurgitation of the neoaortic valve is the major complication of the pulmonary autograft procedure. The incidence of reoperation appears to be lowest with the root replacement technique. Certain conditions (acute rheumatic fever, juvenile rheumatoid arthritis, systemic lupus, ankylosing spondylitis, Libman-Sachs endocarditis, and possibly a dilated aortic root) may be contraindications to the use of a pulmonary autograft. Reoperation on the pulmonary allograft that is used to replace the autograft may be necessary in up to 20% of patients at 20 years. PMID- 10391324 TI - Recurrence of aortic insufficiency after aortic root remodeling with valve preservation. AB - BACKGROUND: Aortic root remodeling (ARR) has recently been proposed for patients with aortic aneurysms and valve insufficiency (AI). To define factors associated with a favorable functional outcome, a review of the mid-term results with ARR was undertaken. METHODS: Between March 1994 and October 1997, 17 consecutive patients (11 men, 6 women), aged 57 +/- 11 years (range 35-71), had elective ARR for aortic aneurysm with or without annuloaortic ectasia (13), sinus of Valsalva aneurysm (3), or chronic aortic dissection (1). Moderate or severe AI was present in 11 patients (65%). Preoperative aortic root diameter was 58 +/- 5 mm (range 51 70). ARR involved replacement of all three aortic sinuses and coronary button reimplantation, using grafts with a mean diameter of 28 +/- 2 mm (range 24-30). RESULTS: There was one early death (6%) due to multiple organ failure. Survivors were followed for 16 +/- 12 months (range 1-44). Actuarial 3-year survival was 94% +/- 6%. Discharge echocardiogram showed a decrease in AI in all patients: AI was absent in 11 (69%) and mild in 5 (31%). Recurrence of moderate or severe AI after a mean of 16 +/- 9 months (range 9-28) was noted in 6 patients (37%), 3 of whom had no AI at discharge. Five of 6 patients required aortic valve replacement. Comparison of demographic and operative variables showed that severe preoperative AI (67% vs 20%, p = 0.06), annuloaortic ectasia (100% vs 20%, p = 0.002), and cystic medial necrosis (100% vs 20%, p = 0.002) were significantly more prevalent in patients developing severe AI at follow-up. The 10 patients (63%) with absent AI showed durable competence of the valve and relief from symptoms at follow-up. CONCLUSIONS: Despite early restoration of valve competence, AI may recur and progress after ARR at medium-term follow-up in a proportion of patients. The severity of preoperative AI and the nature of aortic root disease may negatively influence the durability of repair. Continued observation of results with ARR appears mandatory to identify the appropriate surgical candidates. PMID- 10391325 TI - Mutations of extracellular matrix components in vascular disease. AB - BACKGROUND: Marfan's syndrome (MFS) is characterized by manifestations in the skeletal, ocular, and cardiovascular systems. Dilatation of the aortic root is the hallmark feature in the cardiovascular system. Aortic dilatation is associated with fragmented elastic fibers and accumulation of amorphous matrix elements in the medial layer. This pathology is caused by mutations in fibrillin 1, the major structural component of elastic microfibrils. Fibrillin 1 mutations may affect the assembly or function of the elastic microfibrils or both. To answer this important question, MFS-like mice have been created. METHODS: MFS like mice were generated by homologous gene targeting in embryonic stem cells. Targeting of the mouse fibrillin 1 gene had the dual effect of reducing gene expression 10-fold and of producing an internally deleted protein. RESULTS: Mutant homozygous mice make very small amounts of only mutant fibrillin 1 and die postnatally of MFS-like vascular complications. Histopathological findings include focal fragmentation of elastic fibers and accumulation of amorphous matrix in the aortic media. CONCLUSIONS: A mouse model for the severe form of MFS has been created using the technique of gene targeting. Aside from its clinical value, the model has demonstrated that fibrillin 1 is predominantly involved in the function rather than the assembly of elastic microfibrils. PMID- 10391326 TI - Operative management of Marfan syndrome: The Johns Hopkins experience. AB - BACKGROUND: Doctor Antoine Marfan described the first case of Marfan syndrome in 1896. It was over 50 years later that the development of aortic aneurysms and subsequent rupture was appreciated as the most life-threatening component of the syndrome. METHODS: Doctor Vincent Gott, at our institution, performed the first Bentall procedure for an aneurysm of the ascending aorta in 1976. Since that time, the aortic root has been replaced in 231 Marfan patients. Of this group, 218 patients had a composite graft repair, 11 had an aortic root replacement with a homograft, and 2 patients had valve sparing procedures. There were 168 males and 63 females. Of the total 231 patients, 150 were operated on by Dr Gott. The remaining 81 patients were operated on by 10 other Hopkins surgeons. The average diameter of the ascending aorta was 6.8 cm, with a range from 4.5 to 10. The average aortic diameter of 43 patients who had an ascending aortic dissection was 7.3 cm. Fourteen of these patients had dissection with an aortic diameter of 6.5 cm or less. RESULTS: Among the 198 patients who underwent elective repair, there was no 30-day mortality. Thirty-three patients underwent urgent repair with 2 deaths, yielding a 30-day mortality of 6.1%. The mortality for the entire group of patients was 0.9%. Complications associated with this series of patients included 8 with endocarditis, 7 with thromboembolism, and 4 late coronary dehiscences. Actuarial survival was 88% at 5 years, 81% at 10 years, and 75% at 20 years. Multivariate analysis revealed New York Heart Association classification, male gender and urgent surgery as independent risk factors for mortality. CONCLUSION: Marfan patients with aortic aneurysms can undergo elective surgery with a low operative risk and excellent long-term survival with low morbidity. We feel that elective resection of an aneurysm in a Marfan patient should occur when it approaches a diameter of 5.5 cm. It is essential that a timely diagnosis be made in this group of young patients. PMID- 10391327 TI - Aortic dissection in Marfan's syndrome. AB - BACKGROUND: Aortic dissection is the most frequent cause of premature death in Marfan's syndrome. Low-risk elective surgery of the abnormal aortic root has the potential to prevent this complication. METHODS: We examine genetic, structural, and pathophysiological mechanisms of aortic dissection and discuss the surgical methods used when dissection occurs. RESULTS: Abnormal fibrillin disturbs the functional relationship between blood flow and vascular endothelial cell response (mechanotransduction). Decreased arterial distensibility also decreases aortic wall stress, thereby predisposing to dissection in the weakened arterial wall. Radical root and wall surgery and lifelong beta-blockade are required after aortic dissection. CONCLUSIONS: Detailed lifelong medical and surgical treatment can greatly prolong life in Marfan's syndrome. Elective aortic root replacement is paramount in preventing aortic dissection and avoiding subsequent problems in the distal aorta. PMID- 10391328 TI - Extensive aortic reconstruction for aortic aneurysms in Marfan syndrome. AB - BACKGROUND: Marfan syndrome patients frequently develop aneurysms or dissections involving multiple segments of the aorta, and occasionally require staged replacement of the entire aorta. This study reviews the surgical outcome of patients with Marfan syndrome who underwent extensive aortic reconstruction. Extensive reconstruction is defined as reconstruction of more than two segments of the ascending, arch, descending thoracic, or abdominal aorta. METHODS: From March 1973 to December 1997, 101 patients with Marfan syndrome underwent aortic operation. Twenty-six patients (25.7%) had extensive aortic reconstruction. All 26 patients suffered from aortic dissection: 13 patients had Stanford type A and 13 had type B dissection. Twenty-three patients (88.4%) had annuloaortic ectasia and aortic regurgitation. Surgical procedures included composite valve graft replacement (n = 23, 88.4%), aortic arch reconstruction (n = 15, 57.7%), graft replacement of the descending thoracic aorta (n = 6, 23.1%), and graft replacement of the thoracoabdominal aorta (n = 16, 61.5%). Five patients (19.2%) had total thoracoabdominal aortic replacement, and three patients (11.5%) had replacement of the entire aorta. Twenty-one patients (80.8%) required multiple operations. RESULTS: Follow-up was complete in all patients. The 30-day survival rate was 88.5%. None of the survivors had paraplegia or paraparesis. The overall long-term survival rate was 88.5 +/- 6% at 1 year, and 81.7 +/- 9% at 9 years. CONCLUSIONS: Aortic surgery prolongs survival in patients with Marfan syndrome, and currently there is a relatively low associated morbidity and mortality even for aggressive surgical treatment. PMID- 10391330 TI - Antegrade cerebral perfusion with cold blood: a 13-year experience. AB - BACKGROUND: In 1986 we introduced the technique of antegrade selective perfusion of the brain with cold blood during surgery of the aortic arch. METHODS: Between January 1984 and March 1998, 171 patients (118 males and 53 females) aged 25 to 83 years (mean 56.5 +/- 17), underwent replacement of the transverse aortic arch with the aid of cold blood antegrade selective perfusion. One hundred twenty two patients (71.3%) with chronic lesions were operated on electively; 49 patients (28.6%) were operated on urgently for acute aortic dissection (42 patients) or for a ruptured chronic aneurysm (7 patients). Fifty-one patients (29.8%) had previously undergone a surgical procedure on the thoracic aorta. Mean duration of cardiopulmonary bypass was 121 minutes (range: 65-248); mean duration of cerebral perfusion was 60 minutes (range: 15-90), and mean duration of systemic circulatory arrest circuit was 32 minutes (range: 10-57). The electroencephalogram, routinely recorded, showed disappearance of electrical activity in a mean of 9 minutes (range: 3-16) initial return of electrical activity after a mean of 12 minutes (range: 1-35) and normalization in a mean time of 66 minutes. RESULTS: All patients but 7 (4%) showed signs of normal awakening within 8 hours postoperatively. Six patients (3.5%) had fatal neurologic complications, and 16 patients (9.3%) had a non-fatal neurologic complications. Twenty-nine patients (16.9%) died during the postoperative hospital course. There was a significant difference between patients aged less than 60 years (9%) and patients older than 60 years (mortality rate 26.4%, p < 0.02). There was also a significant difference between patients undergoing an isolated replacement of the arch, and those in whom the replacement was extended to the descending aorta in whom mortality was 36.4% (chi2, p < 0.02). Lesion and gender had no significant influence on the outcome of the patients, nor had the duration of cardiopulmonary bypass, circulatory arrest, and cerebral perfusion. In particular, no correlation could be established between the duration of cerebral perfusion and the occurrence of neurologic complications. CONCLUSION: The clinical results obtained throughout this experience have demonstrated that selective antegrade cerebral perfusion with cold blood provides excellent protection during surgery of the transverse aortic arch. In addition, it avoids the use of deep hypothermia and prolonged cardiopulmonary bypass and does not limit the time allowed to perform the aortic repair. In our opinion it is the technique of choice, especially in frail patients or those requiring a long and difficult procedure. PMID- 10391329 TI - Assessing the impact of cerebral injury after cardiac surgery: will determining the mechanism reduce this injury? AB - BACKGROUND: Central nervous system dysfunction continues to produce significant morbidity and associated mortality in patients undergoing cardiac surgery. Using a closed-chest canine cardiopulmonary bypass model, dogs underwent 2 h of hypothermic circulatory arrest (HCA) at 18 degrees C, followed by resuscitation and recovery for 3 days. Animals were assessed functionally by a species-specific behavioral scale, histologically for patterns of selective neuronal necrosis, biochemically by analysis of microdialysis effluent, and by receptor autoradiography for N-methyl-D-aspartate (NMDA) glutamate receptor subtype expression. RESULTS: Using a selective NMDA (glutamate) receptor antagonist (MK801) and an AMPA antagonist (NBQX), glutamate excitotoxicity in the development of HCA-induced brain injury was documented and validated. A microdialysis technique was employed to evaluate the role of nitric oxide (NO) in neuronal cell death. Arginine plus oxygen is converted to NO plus citrulline (CIT) by the action of NO synthase (nNOS). CIT recovery in the cerebrospinal fluid and from canine cortical homogenates increased during HCA and reperfusion. These studies demonstrated that neurotoxicity after HCA involves a significant and early induction of nNOS expression, and neuronal processes leading to widespread augmentation of NO production in the brain. To further investigate the production of excitatory amino acids in the brain, we hypothesized the following scenario: HCA--> increased glutamate, increased aspartate, increased glycine--> increased intracellular Ca2+--> increased NO + CIT. Using the same animal preparation, we demonstrated that HCA caused increased intracerebral glutamate and aspartate that persists up to 20 h post-HCA. HCA also resulted in CIT (NO) production, causing a continued and delayed neurologic injury. Confirmatory evidence of the role of NO was demonstrated by a further experiment using a specific nNOS inhibitor, 7-nitroindazole. Animals underwent 2 h of HCA, and then were evaluated both physiologically and for NO production. 7-Nitroindazole reduced CIT (NO) production by 58.4 +/- 28.3%. In addition, dogs treated with this drug had superior neurologic function compared with untreated HCA controls. CONCLUSIONS: These experiments have documented the role of glutamate excitotoxicity in neurologic injury and have implicated NO as a significant neurotoxin causing necrosis and apoptosis. Continued research into the pathophysiologic mechanisms involved in cerebral injury will eventually yield a safe and reliable neuroprotectant strategy. Specific interventional agents will include glutamate receptor antagonists and specific neuronal NO synthase inhibitors. PMID- 10391331 TI - Retrograde cerebral perfusion for aortic arch surgery: analysis of risk factors. AB - BACKGROUND: Retrograde cerebral perfusion (RCP) has been widely adopted during aortic arch surgery under hypothermic circulatory arrest (HCA). However, the risks in terms of mortality and morbidity in aortic arch surgery using HCA with RCP have not yet been confirmed. METHODS: The present study is a retrospective review of 249 patients who underwent aortic arch surgery at three Japanese cardiovascular centers where RCP is a routine adjunct. The median age was 65 years, and 38 patients were more than 75 years old. The pathology in the aortic arch was atherosclerotic aneurysm in 133 patients and dissection in 116. Seventy patients had surgery on an emergency basis. Surgery was performed through a median sternotomy in 182 patients and through a left thoracotomy in 67. Using HCA with RCP, graft replacement of the total aortic arch was performed in 109, the distal arch in 63, and the ascending aorta and hemi-arch in 66; 11 patients had patch repair. RESULTS: The overall hospital mortality was 25/249 (10%), and 12/70 (17%) in emergent surgery. Stroke developed in 11 patients (4%). The median duration of RCP was 46 minutes (range, 5 to 95). Univariate analysis of risk factors revealed that an age of 75 years or more (p < 0.001), and urgency of surgery (p = 0.02) affected hospital mortality. Multivariate logistic analysis revealed that pump time (p = 0.0001), age (p = 0.0001) and RCP time (p = 0.05) are the most significant risk factors for mortality. The risk factors for mortality and neurological morbidity combined are pump time (p = 0.0001), age (p = 0.0002), and urgency of surgery (p = 0.07); RCP time is marginally significant (p = 0.15). CONCLUSIONS: The dominant risk factors for mortality and morbidity are pump time, urgency of the surgery, and age. RCP is a simple and useful adjunct for aortic arch surgery with up to 80 minutes of HCA, although prolonged RCP is a risk factor for mortality and morbidity. PMID- 10391332 TI - Risk analysis for aortic surgery using hypothermic circulatory arrest with retrograde cerebral perfusion. AB - BACKGROUND: Retrospective analysis of 144 patients undergoing aortic arch reconstruction using hypothermic circulatory arrest (HCA) with retrograde cerebral perfusion (RCP) for cerebral protection was performed. METHODS: The diagnosis, procedure, and anatomic site of the arch anastomosis were analyzed to see if they were independent predictors of mortality or morbidity. In addition age, gender, HCA-RCP times, preoperative malperfusion (both treated and untreated), surgical status, and redo surgery status were also examined to determine their influence on the incidence of death and complications. Both multivariate and univariate analysis were performed using linear regression and cross-tabulation with either chi2 or Fisher's exact test where appropriate. RESULTS: Preoperative surgical status (emergent) and the presence of untreated preoperative malperfusion were the only variables that were significant independent predictors for mortality (p <0.05). No variable was significant for the prediction of stroke or other complications. The severity of surgery had no bearing on the patient outcome. CONCLUSIONS: Complex aortic surgery using HCA-RCP can be performed with acceptable risk to the patients. PMID- 10391333 TI - Temporary neurological dysfunction after deep hypothermic circulatory arrest: a clinical marker of long-term functional deficit. AB - BACKGROUND: With increasing clinical experience, it has become clear that two distinct forms of neurological injury occur after operations on the thoracic aorta that require temporary exclusion of the cerebral circulation. Traditionally, evaluation of neurological outcome was limited to reporting the incidence of postoperative stroke related to ischemic infarcts due to particulate embolization. More recently, the symptom complex defined as "temporary neurological dysfunction" (TND) was recognized as a functional manifestation of subtle and presumably transient brain injury, but whether this early postoperative syndrome is associated with long-term deficits of cognitive and intellectual functions has not been established. METHODS: With Institution Review Board approval, 105 patients undergoing elective thoracic aortic surgery were entered into a protocol involving neuropsychological evaluation with a battery of tests preoperatively, and 1 and 6 weeks postoperatively. Patients who could not be tested adequately or had documented strokes were eliminated from final analysis. Seventy-one patients completed the neuropsychological evaluation, which consisted of eight tests consolidated into five domains: attention, cognitive speed, memory, executive function, and fine motor function. Independent observers also determined whether temporary dysfunction was present, and graded its severity based on a fixed but subjective clinical scale, ranging from simple disorientation and lethargy or confusion (grade 1-2) to prolonged extreme agitation or psychotic behavior requiring treatment with psychotropic drugs (grade 3-5). Data were normalized to baseline values, and were analyzed using analysis of variance, analysis of covariance (ANCOVA), and chi2 as necessary. RESULTS: A previous analysis had shown that patients who could not be tested or had poor scores 1 week postoperatively were more likely to perform poorly at 6 weeks (odds ratio 5.27, p < 0.01). In the current study, in order to determine the clinical relevance of TND, patients were analyzed retrospectively according to their performance in neuropsychological testing: patients with no change or a decline of less than 50% in tests of memory, motor function, and attention 1 week postoperatively (group 1, n = 49) were compared with those with a negative change exceeding 50% in the same functions at 1 week (group 2, n = 22). The overall incidence of TND was 28.1% (20/71). The incidence of TND in group 2 (14/22, 63%) was significantly higher than in group 1 (6/49, 12%; p = 0.0006). Similarly, the severity of TND (as assessed by clinical score > 2) was also significantly higher in group 2 (11/14) compared with group 1 (0/6; p = 0.006.) CONCLUSIONS: The incidence and severity of clinically apparent temporary neurological dysfunction correlates significantly with poor performance on neuropsychological tests 1 week postoperatively. Such poor performance predicts continued deficits in memory and motor function at 6 weeks. Thus, TND may not be a benign self-limited condition as previously supposed, but rather a clinical marker for insidious but significant neurological injury associated with measurable long-term deficits in cerebral function. A concerted effort to reduce the incidence of this complication is therefore necessary. PMID- 10391334 TI - Cerebral metabolic suppression during hypothermic circulatory arrest in humans. AB - BACKGROUND: Hypothermic circulatory arrest (HCA) is used in surgery for aortic and congenital cardiac diseases. Although studies of the safety of HCA in animals have been carried out, the degree to which metabolism is suppressed in patients during hypothermia has been difficult to determine because of problems with serial measurements of cerebral blood flow in the clinical setting. METHODS: To quantify the degree of metabolic suppression achieved by hypothermia, we studied 37 adults undergoing operations employing HCA. Cerebral blood flow was estimated using an ultrasonic flow probe on the left common carotid artery, and cerebral arteriovenous oxygen content differences were calculated from jugular venous bulb and arterial oxygen saturations. Cerebral metabolic rates while cooling were then ascertained. The temperature coefficient, Q10, which is the ratio of metabolic rates at temperatures 10 degrees C apart, was determined. RESULTS: The human cerebral Q10 was found to be 2.3. The cerebral metabolic rate is still 17% of baseline at 15 degrees C. If one assumes that cerebral blood flow can safely be interrupted for 5 min at 37 degrees C, and that cerebral metabolic suppression accounts for the protective effects of hypothermia, the predicted safe duration of HCA at 15 degrees C is only 29 min. CONCLUSIONS: The safe intervals calculated from measured cerebral oxygen consumption suggest that shorter intervals and lower temperatures than those currently used may be necessary to assure adequate cerebral protection during hypothermic circulatory arrest. PMID- 10391335 TI - Does retrograde cerebral perfusion affect risk factors for stroke and mortality after hypothermic circulatory arrest? AB - BACKGROUND: In aortic surgery requiring hypothermic circulatory arrest (HCA), retrospective studies identify age and duration of the arrest period as predictors of stroke and mortality. Retrograde cerebral perfusion (RCP) has been reported to reduce the risk of stroke when compared with historical controls. The aim of this study was to ascertain if RCP affected mortality, stroke, or the risk factors for these end points in a consecutive series of HCA patients. METHODS: We investigated the impact of RCP in 130 patients, mean age 62.7 years (range 20 84); 78 were men and 35% were emergencies. Overall mortality was 16.9% (elective 9.5%) and the incidence of stroke was 6.9%. Mean HCA time was 30.1 min (95% confidence interval [CI] 27.9-34). RCP was instituted in 96 cases for a mean of 24.4 min (95% CI 21.9-27.0). RESULTS: Perioperative univariate predictors of mortality were emergency status, acute rupture, long HCA and cardiopulmonary bypass duration, and postoperative complications. For stroke, age (p = 0.007), hypertension (p = 0.05), and long HCA duration (p = 0.01) were predictive. RCP did not decrease mortality (p = 0.18, OR 0.55) or incidence of stroke (p = 0.57, OR 1.26). Mortality after stroke was 44.4% (p = 0.04, OR 4.6). Multiple logistic regression showed severe aortic atherosclerosis and RCP duration (p = 0.038) as risk factors for mortality, and myocardial ischemic time (p = 0.012) and HCA duration (p = 0.05) as risk factors for stroke. HCA and RCP groups differed in HCA duration (HCA mean 25 min [10-80], RCP mean 32 min [10-69]; p < 0.019). CONCLUSIONS: Age and HCA duration remain risk factors for stroke and mortality despite RCP. However, HCA times were longer in the RCP patients, and the patients were not randomized. The role of RCP in cerebral protection requires further prospective randomized studies. PMID- 10391336 TI - Antegrade selective cerebral perfusion in operations on the proximal thoracic aorta. AB - BACKGROUND: To determine the factors that influence hospital death and neurologic complications after surgery on the thoracic aorta using circulatory arrest and antegrade selective cerebral perfusion. METHODS: From May 1989 through April 1997, 106 patients underwent surgery on the thoracic aorta using circulatory arrest and antegrade selective cerebral perfusion. Mean age was 64.0 +/- 11.5 years. Unilateral antegrade cerebral perfusion was used in 37 patients (35%), bihemispheric antegrade cerebral perfusion in 69 patients (65%). Mean antegrade cerebral perfusion time was 50.5 +/- 20.5 minutes. Indication for surgery was atherosclerotic aneurysm in 60 (56.5%) patients, postdissection aneurysm in 26 (24.4%), acute type A dissection in 16 (15.1%), other in 4 (4.0%). RESULTS: Hospital mortality was 8.5% (n = 9; 70% CL: 5.8%-11.2%). Independent predictors of hospital mortality were rethoracotomy (odds ratio 5.7, p = 0.02), postoperative temporary (odds ratio 17.3, p = 0.02) or permanent (odds ratio 7.5, p = 0.03) neurologic dysfunction, postoperative dialysis (odds ratio 9.9, p = 0.008). Bilateral antegrade selective cerebral perfusion had a favorable impact on hospital mortality (odds ratio 0.08, p = 0.007). Temporary neurologic dysfunction occurred in 3.8% of patients (n = 4; 70% CL: 2.0%-5.6%); preoperative hemodynamic instability (odds ratio 14.8, p = 0.05) and perioperative technical problems (odds ratio 22.2, p = 0.033) were independent determinants of temporary neurologic dysfunction. Permanent central neurologic damage occurred in 5.4% of patients (n = 6; 70% CL: 3.2%-7.6%). Preoperative hemodynamic instability (odds ratio 18.9, p = 0.009) and approach through a left thoracotomy (odds ratio 9.4, p = 0.031) were significant predictors of permanent neurologic damage. CONCLUSIONS: Hospital mortality is affected significantly by the choice of technique used for antegrade cerebral perfusion. The incidence of both temporary and permanent postoperative central neurologic damage is influenced by preoperative hemodynamic instability. Duration of cerebral perfusion had no influence on the postoperative neurologic outcome. PMID- 10391337 TI - S-100beta release in hypothermic circulatory arrest and coronary artery surgery. AB - BACKGROUND: Aortic surgery utilizing profound hypothermic circulatory arrest (HCA) has a higher incidence of neurological injury than coronary artery bypass grafting (CABG). S-100beta is a potential marker of cerebral ischemic injury. The aim of this study is to assess its use in investigating cerebral injury during HCA. METHODS: We studied 40 patients (10 CABG, 30 HCA). The mean cardiopulmonary bypass (CPB) times were 72 and 158 minutes, respectively. Mean HCA duration was 27.6 min, with retrograde cerebral perfusion (RCP) used in 18 patients (mean 28.5 minutes, 95% CI 16-25). Perioperative venous blood samples were subjected to S100beta assay. RESULTS: S100beta levels with HCA (peak: 2.68 microg/L, 95% CI 1.99-3.38 microg/L; calculated area under the curve [AUC]: 1596 microg/L/min, 95% CI 825-2368 microg/L/min) were significantly higher (peak, p = 0.028 and AUC, p = 0.007) than with CABG (peak: 1.16 microg/L, 95% CI 0.25-2.1 microg/L and AUC: 53.4 microg/L/min 95% CI 3.0-103.8). Peak S100beta correlated with CPB time in CABG cases (r = 0.76, p < 0.05), and with both CPB and HCA time in HCA cases: without RCP (r = 0.46 and 0.21, respectively, p > 0.05) and with RCP (r = 0.88 and 0.33, respectively, p < 0.05). There was no significant difference in the S100beta levels between HCA groups with and without RCP, but HCA time was longer in the RCP group (p = 0.05). CONCLUSIONS: S100beta release correlates with duration of CPB and HCA. Elevated serum S100 indicates astrocyte death or activation, and suggests blood-brain barrier dysfunction. The continuing release of S100 after the end of operation suggests that HCA may be associated with greater injury than CABG. RCP did not influence S-100beta release in this study. PMID- 10391339 TI - Surgical intervention criteria for thoracic aortic aneurysms: a study of growth rates and complications. AB - BACKGROUND: Evidence regarding the behavior of thoracic aortic aneurysm (TAA) is limited. This study reviews our ongoing efforts to understand the factors influencing aortic growth rates and the complications of rupture and dissection in order to define scientifically sound criteria for surgical intervention. METHODS: Data from 370 patients with TAA treated at Yale University School of Medicine from January 1985 to June 1997 were analyzed. This computerized data base included 1063 imaging studies (magnetic resonance imaging, computed tomography, and echocardiography). RESULTS: The mean size of the thoracic aorta in these patients at initial presentation was 5.2 cm (range 3.5-10). The mean growth rate was 0.10 cm/year. Median size at the time of rupture or dissection was 5.9 cm for ascending and 7.2 cm for descending aneurysms. The incidence of dissection or rupture increased with aneurysm size. Multivariable regression analysis to isolate risk factors for acute dissection or rupture revealed that size > or = 6.0 cm increased the probability of these devastating complications by 25.2% for ascending aneurysms (p = 0.006 compared with aneurysms 4.0-4.9 cm). For descending aneurysms > or = 7.0 cm, risk of dissection or rupture was increased by 37.3% (p = 0.031). CONCLUSIONS: If the median size at time of dissection or rupture had been used as the indication for intervention, half the patients would have suffered a devastating complication before surgery. Accordingly, a criterion lower than the median is appropriate. We recommend 5.5 cm as an acceptable size for elective resection of ascending aortic aneurysms because this operation can be performed with relatively low mortality. For aneurysms of the descending aorta, where perioperative complications are greater and the median size at the time of complication is larger, we recommend intervention at 6.5 cm. PMID- 10391338 TI - Retrospective study of somatosensory evoked potential monitoring in deep hypothermic circulatory arrest. AB - BACKGROUND: We evaluated the efficiency of median-nerve somatosensory evoked potentials (SEPs) monitoring in determining the optimal level of hypothermia in 62 consecutive patients operated on under deep hypothermic circulatory arrest (CA). METHODS: CA was started at 1 degree C below the temperature at which both brainstem and cortical SEP components disappear. No additional method of cerebral protection was used. RESULTS: New neurological complications were observed in 15 patients: long-lasting in 11 and transient in 4. A retrospective analysis of SEP monitoring identified the origin of the complications in 12 cases: early CA with incomplete cooling due to emergency (3 cases); inefficient retrograde perfusion through the femoral artery during cooling due to the dissection flap effect (4 cases); embolism during rewarming (2 cases); delayed embolism (2 cases); hemorrhagic shock (1 case). In 2 cases, neurological sequelae involved the lower limbs (extracerebral origin). One case without intraoperative SEP changes was neurologically abnormal preoperatively and did not change postoperatively. There were no cases with sequelae due to excessive CA duration. CONCLUSIONS: The use of SEP monitoring to determine the optimal level of hypothermia efficiently prevents neurological sequelae of CA. It helps in monitoring the degree of cerebral protection during cooling (flap effect), and rewarming. PMID- 10391340 TI - Natural history of descending thoracic and thoracoabdominal aneurysms. AB - BACKGROUND: A review of 165 patients with chronic dissecting and degenerative aneurysms of the descending thoracic and thoracoabdominal aorta initially managed nonoperatively was carried out to ascertain factors associated with a high risk of rupture. METHODS: Changes in the aneurysms were followed with three dimensional reconstructions of computed tomograph scans. Risk factors were compared in patients with dissecting and nondissecting aneurysms who experienced rupture, in whom operation was recommended during the course of follow-up, and in those without rupture or operation. RESULTS: Nondimensional variables associated with an enhanced risk of rupture include age, the presence of chronic obstructive pulmonary disease, and even uncharacteristic continued pain. Patients with rupture of dissections had significantly higher blood pressures than survivors, and significantly smaller maximal descending thoracic aortic diameters (median 5.4 cm) than patients with rupture of degenerative aneurysms (median 5.8 cm). The extent of the aneurysm, as reflected by the maximal abdominal aortic diameter, was a significant risk factor for rupture only in nondissecting aneurysms. Mortality from rupture was significantly higher in patients with chronic dissections than in patients with nondissecting aneurysms: 9/10 vs 26/34 (p = 0.004). CONCLUSIONS: Almost 20% of patients followed nonoperatively succumbed to rupture, suggesting that a more aggressive surgical approach toward patients with chronic aneurysms of the descending thoracic and thoracoabdominal aorta is warranted. An individualized risk of rupture within 1 year can now be calculated, and patients whose operative risk is lower than their calculated risk should be offered elective surgery. PMID- 10391341 TI - Left heart bypass reduces paraplegia rates after thoracoabdominal aortic aneurysm repair. AB - BACKGROUND: The optimal strategy for spinal cord protection during thoracoabdominal aortic aneurysm (TAAA) repair remains unclear. We evaluated the protective effect of left heart bypass (LHB) during repair of extensive TAAAs. METHODS: During a 12-year period, 710 patients had repair of extent I or II TAAAs. Left heart bypass was used in 312 (43.9%) patients. This group was retrospectively compared with 398 (56.1%) patients who had operations without LHB. RESULTS: The overall 30-day survival rate was 94.8% (673 patients). In 42 patients, (6.0%) paraplegia or paraparesis developed. In patients with extent I TAAAs, paraplegia and paraparesis rates in LHB (6 of 123, 4.9%) and non-LHB (9 of 246, 3.7%) groups were similar (p = 0.576) despite longer aortic clamp times in the former group. In patients with extent II TAAAs, the LHB group had a lower incidence of paraplegia or paraparesis (9 of 189, 4.8%) compared with the non-LHB group (18 of 137, 13.1%; p = 0.007). CONCLUSIONS: Left heart bypass reduced the risk of paraplegia and paraparesis in patients who had repair of extent I and II TAAAs. PMID- 10391342 TI - An approach to spinal cord protection during descending or thoracoabdominal aortic repairs. AB - BACKGROUND: During the past few years, after much research, progress has been made in reducing the risk of spinal cord injury after descending or thoracoabdominal aortic repairs. METHODS: Based on that research I describe a method to reduce the risk of spinal cord injury. RESULTS: Our data show that with this technique, less than 5% of our patients had a permanent injury whereby they are unable to walk. CONCLUSION: The use of intrathecal papaverine, cerebrospinal fluid drainage, hypothermia, and reimplantation of intercostal arteries from below T6 to and including L1 are recommended. PMID- 10391343 TI - Spinal cord protection in descending thoracic and thoracoabdominal aortic repair. AB - BACKGROUND: During simple cross-clamp repair of the descending thoracic or thoracoabdominal aorta, the likelihood of neurologic complications increases greatly after only 30 minutes of spinal cord ischemia. At greatest risk are patients with type II thoracoabdominal aortic aneurysms. METHODS: We reviewed our experience of simple cross-clamp repair and procedures accompanied by adjuncts, paying particular attention to the outcome of patients who had type II thoracoabdominal aortic aneurysms. Between February 1991 and March 1998, 508 patients had descending thoracic and thoracoabdominal aortic repair, 255 (50.2%) of whom received the adjuncts of cerebrospinal fluid drainage and distal aortic perfusion. RESULTS: Fifteen patients died on the day of operation and could not be evaluated for neurologic deficit. The overall incidence of neurologic deficit was 33 of 493 patients (6.7%). In patients who received adjuncts, neurologic deficit occurred in 9 of 247 (3.6%) overall; in types I and II it was 8 of 164 (4.9%), and in type II alone, 7 of 87 (8.1%). Neurologic deficit in simple cross clamp patients was 24 of 246 (9.8%) overall; in types I and II it was 15 of 99 (15.2%), and in type II alone, 13 of 44 (29.6%). CONCLUSIONS: With the surgical adjuncts of cerebrospinal fluid drainage and distal aortic perfusion, the probability of neurologic deficit is lowered appreciably. PMID- 10391344 TI - Hypothermic cardiopulmonary bypass for spinal cord protection: rationale and clinical results. AB - BACKGROUND: Hypothermic cardiopulmonary bypass with or without circulatory arrest has been used successfully for the treatment of complex aneurysms of the descending thoracic and thoracoabdominal aorta. Hypothermia has a protective effect on spinal cord function, and its use has been associated with a low incidence of paraplegia in traditionally high-risk patients. Experimentally, the protective effect of hypothermia has been related to amelioration of excitotoxic injury by reduction of neurotransmitter release and to inhibition of delayed apoptotic cell death. METHODS: During a 12-year period, 114 patients with descending thoracic or thoracoabdominal aortic disease underwent replacement of the involved aortic segments using hypothermic cardiopulmonary bypass and intervals of circulatory arrest. RESULTS: The hospital mortality was 8% (9 patients). Paraplegia occurred in 2 and paraparesis in 1 of the 108 patients whose lower limb function was assessed postoperatively (2.8%). None of 40 patients with aortic dissection and none of the last 81 patients in the series developed paralysis. CONCLUSIONS: Our experience with hypothermic cardiopulmonary bypass and circulatory arrest confirms that hypothermia provides substantial protection against paraplegia, and it allows complex operations on the descending thoracic and thoracoabdominal aorta to be performed with acceptable mortality. PMID- 10391345 TI - Sensitivity, specificity, and surgical impact of somatosensory evoked potentials in descending aorta surgery. AB - BACKGROUND: We evaluate the efficiency of multilevel somatosensory evoked potential (SEP) monitoring for intraoperative re-establishment of blood flow to the spinal cord in 63 patients undergoing descending aorta repair. METHODS: The presence of critical vessels in a cross-clamped aortic segment was ascertained by a 15 minute SEP observation period while the segment between the clamps was vented to drain out the collateral flow. RESULTS: SEPs influenced the surgical strategy in 17 cases (27%): use of the Biomedicus in 1 traumatic rupture; critical vessel reimplantation or distal clamp replacement in 13 cases of segmental spinal ischemia; and hastening the procedure or proximal clamp replacement in 3 cases of left carotid ischemia. There were no cases of unexplained multilevel SEP abnormalities. Immediate paraplegia was observed in 2 cases (1 pre-existing; 1 forecast by a 199-minute period of SEP absence due to segmental ischemia); 2 patients presented delayed paraplegias despite unchanged intraoperative SEPs, and 1 case presented a transient paraplegia due to lower motoneuronal involvement. CONCLUSIONS: SEPs efficiently identified critical vessels to be reimplanted in order to avoid immediate paraplegia. However, systematic additional vessel reimplantation, if technically feasible, and prolongation of SEP monitoring during the postoperative period with careful blood pressure control are needed to prevent delayed paraplegia. PMID- 10391346 TI - Use of somatosensory evoked potentials for thoracic and thoracoabdominal aortic resections. AB - BACKGROUND: Despite tremendous development in surgical and anesthetic techniques, resection of the thoracic and thoracoabdominal segments of the aorta remain associated with the risk of paralysis. Routine use of somatosensory-evoked potential (SEP) monitoring in patients undergoing surgery of the thoracic aorta has become a standard intra- and postoperative procedure at our institution since its first use in 1993. METHODS: One hundred forty nine (149) thoracic aortic operations were performed during January 1993 through January 1998 using SEP directed serial sacrifice of paired intercostal arteries. Full, partial, or no cardiovascular bypass was variably used, dictated by anatomy; 49 patients required deep hypothermic circulatory arrest (DHCA). Patients were monitored during both the intraoperative procedure as well for the post-anesthesia period until neurologic stability and/or ability to reproducibly demonstrate lower extremity neurologic competency was established. Postoperative neurologic function was compared to ischemic intervals, extent of aortic resection, number of intercostal arteries sacrificed, type of perfusion, and underlying aortic pathology. RESULTS: Overall mortality in the group was 13 patients (8.7%), with no one cause predominating. Nine patients sustained permanent paraplegia, only 1 of whom lost SEPs during the procedure. Abnormal SEPs were seen in 19 patients, 14 of whom had normal neurologic function after awakening. Three of 19 (15.8%) developed late paraplegia that resolved with medical therapy. Eleven patients (7.4%) developed cerebrovascular accidents (CVA), with the majority (8) appearing in the group undergoing DHCA. The risk of CVA was significantly higher in DHCA patients (p < 0.01) than other patients. No patient with CVA had abnormal SEPs; 4 DHCA patients developed abnormal SEPs, 1 with permanent paralysis. CONCLUSIONS: The routine use of SEP monitoring during thoracic and thoracoabdominal aortic surgery as well as during the postoperative period may be useful in decreasing the observed incidence of paraplegic events associated with these procedures. PMID- 10391347 TI - Aortic aneurysm operations: past, present, and future. AB - Effective methods to treat aortic aneurysms are now available, although these lesions still challenge the cardiovascular surgeon. Attempts at treatment began in earnest in the 1800s, with the introduction of indirect and direct methods of repair. A major breakthrough occurred in the late 1800s, when Dr Rudolph Matas devised a method for internal repair of aneurysms in which continuity of blood flow was restored by excising the diseased portion of the lesion and creating a tunnel through the remaining normal portion. Matas named this technique reconstructive endoaneurysmorrhaphy. Until that time, surgeons had treated aneurysms by ligating the parent vessel with a Hunterian ligature or introducing foreign material to promote coagulation. Ligating the aneurysm rendered the extremities vulnerable to ischemic damage, however, and results were unpredictable with the use of various foreign materials. Around the turn of the century, Carrel began experimenting with different techniques for vascular anastomoses. The work of these early pioneers formed the basis for much of the modern treatment of aneurysms of the thoracic aorta. My experience began in 1950, when I excised a large aortic aneurysm in one of Dr Alfred Blalock's patients. The patient survived and was cured. After that experience, I knew that aortic aneurysms could be treated successfully by aggressive surgical means. Treatment has changed, however, from the early emphasis on excising the lesion to the present practice of restoring circulatory continuity with a suitable graft, ie, endoaneurysmorrhaphy. The development of reliable synthetic grafts has been one of the most important advances in the treatment of aneurysms. The surgical technique used today depends on the anatomic location of the aneurysm, which can occur anywhere from the aortic annulus and aortic valve to the distal thoracic aorta and visceral vessels in the abdomen. PMID- 10391348 TI - Impact of left heart bypass on the results of thoracoabdominal aortic aneurysm repair. AB - BACKGROUND: This study evaluated the role of left heart bypass on the results of thoracoabdominal aortic aneurysm (TAAA) operations. METHODS: Two hundred fifty eight patients had surgical repair of a thoracoabdominal aortic aneurysm between 1981 and 1998 using the inlay technique. Simple cross-clamping was used in 47.7% and left heart bypass (atriodistal) in 52.3%. Further surgical technique was identical: liberal intercostal or lumbar artery reimplantation, cerebrospinal fluid drainage (since 1989), administration of a renal cooling solution, permissive mild hypothermia, and no pharmacologic protection. Both univariate and multivariate analysis were used. RESULTS: The hospital mortality rate was 10.1% overall: 14.6% in the cross-clamp group, and 5.9% in the bypass group (p = 0.02). The risk of hospital death increased with aneurysm rupture (odds ratio 5.6) and when the patient needed postoperative dialysis (odds ratio 7.5). The use of left heart bypass had a mild protective effect on hospital death (odds ratio 0.56). The incidence of postoperative renal failure requiring dialysis was 8.3% overall: 10.9% in the cross-clamp group, and 5.9% in the bypass group (p = 0.16). After multivariate analysis, a longer operative procedure (odds ratio 1.01 per minute) and a longer reappearance time of blue dye in the urine (odds ratio 1.05 per minute) increased the risk of dialysis, whereas the use of atriodistal bypass reduced that risk (odds ratio 0.08). Paraplegia or paraparesis occurred in 10.9% of patients overall: 13.2% in the cross-clamp group, and 8.8% in the bypass group (p = 0.27). After logistic regression, rupture increased the risk of paraplegia or paraparesis (odds ratio 3.2) and dissection reduced it (odds ratio 0.23). CONCLUSIONS: The use of atriodistal bypass is beneficial in patients who had thoracoabdominal aortic aneurysm repair. Hospital mortality rates, postoperative dialysis, and paraplegia/paraparesis were reduced. PMID- 10391349 TI - Prediction of thoracic aortic aneurysm expansion: validation of formulae describing growth. AB - BACKGROUND: The expansion rate of thoracic aortic aneurysms may be an important and clinically relevant index of the risk of rupture. The aims of this study were to assess the validity of three published exponential equations that predict expansion rate in a separate sample population, and to calculate an expansion rate formula for this cohort of patients. METHODS: We studied 88 consecutive patients undergoing serial computed tomographic or magnetic resonance imaging scanning to monitor thoracic aortic aneurysm progression. In interval scans of at least 6 months, we measured minimum coronal aortic diameter at seven set levels and maximal diameter, yielding 780 segment-intervals. RESULTS: The linear expansion rate (mean 2.6 mm/year) increased with incremental aortic diameter (aortic diameter < 40 mm: 2.0; 40-49 mm: 2.3; 50-59 mm: 3.6; > or = 60 mm: 5.6 mm/year; p < 0.01). Regression analysis showed close correlation between predicted and sample data, but there were significant differences between observed and expected measurements. The Yale formula underestimated growth by 0.8 mm, while Mt. Sinai and Osaka formulae overestimated actual change by 1.5 and 0.2 mm, respectively. The expansion rate derived from our population was: last diameter = initial diameter x e(0.00367 x time) (r = 0.617). CONCLUSIONS: Although formulae derived from one thoracic aortic aneurysm sample population may not extrapolate exactly to others, there is close concordance of results for patient populations in three different continents. PMID- 10391350 TI - Thoracic aortic aneurysm repair with an endovascular stent graft: the "first generation". AB - OBJECTIVE: The feasibility and efficacy trial of an endovascular stent-grafting system for the treatment of aneurysms of the descending thoracic aorta was investigated. METHODS: After Institutional Review Board approval, 103 patients (mean age 69 years) underwent stent graft repair of a descending thoracic aortic aneurysm between July 1992 and November 1997. The stent graft was fabricated using self-expanding "Z" stents covered by a woven Dacron tube graft. Follow-up, which averaged 22 months, was 100% complete. Simultaneous open abdominal aortic aneurysm repair was performed in 19 patients. RESULTS: Complete aneurysm thrombosis was achieved in 86 patients (83%). Early mortality, defined as a death during the same hospitalization or in less than 30 days, was 9 +/- 3%, and was significantly associated with preoperative cerebrovascular accident (CVA) or myocardial infarction. Major perioperative morbidity occurred in 31 patients, and included paraplegia in 3, CVA in 7, and respiratory insufficiency in 12 patients each. Actuarial survival was 81 +/- 4% at 1 year, and 73 +/- 5% at 2 years. Treatment failure (including all late, sudden, unexplained deaths) occurred in 38 patients, and only 53 +/- 10% of patients were free of treatment failure at 3.7 years. Five patients required late operative therapy for endoleaks associated with aneurysm enlargement. CONCLUSIONS: Satisfactory results were achieved using this "first-generation" homemade stent graft device. Mortality and morbidity occurred frequently, but may have been associated with the high-risk character of this patient population. Medium-term results were acceptable, but continued aortic enlargement, with the late development of endoleaks, is a significant concern. Second-generation devices with commercial development, coupled with this initial experience, should allow improved clinical results in the future. Longer term follow-up is still necessary to fully define the efficacy of this endovascular approach. PMID- 10391351 TI - Subtypes of acute aortic dissection. AB - BACKGROUND: This series consists of a 12-year experience with a policy of identifying and replacing the aortic segment containing the primary intimal tear for repair of acute aortic dissection. METHODS: Patients with type A dissection underwent urgent surgery. Patients with type B dissection were referred for surgery based on selective criteria, including aortic dilatation greater than 5 cm. A classification system for acute dissection is described that specifies the site of intimal tear while retaining the clinical relevance of the Stanford system. RESULTS: Of 168 acute dissections, 139 were type A and 29 were type B. The site of intimal tear was as follows: ascending aorta, 83 cases; arch, 32 cases; descending aorta, 29 cases; multiple tears, 11 cases (10 included arch tears); no tear (intramural hematoma), 6 cases; not noted, 7 cases. Only 60% of acute type A dissections arose from solitary intimal tears in the ascending aorta, whereas 30% had arch tears. Hospital mortality for type A dissection was 13.7% (18.8% for arch tears, NS) and 0% for type B. False lumen patency was 57.1% for type A dissection and 18.8% for type B dissection (p = 0.002), yet survival was similar for these groups. Ten-year survival for type A dissection with arch tear (0.51 +/- 0.12) was lower than 10-year survival for type A dissection with ascending tear (0.74 +/- 0.05; p = 0.77), and significantly lower than for type A dissection with descending tear (0.88 +/- 0.12; p = 0.029). CONCLUSIONS: Systematic resection of the primary tear yielded similar hospital mortality, 5 year survival, and aorta-related event-free survival rates for subtypes of acute type A dissection. Excellent results were obtained with a selective approach to type B dissection. PMID- 10391352 TI - How to obtain hemostasis after aortic surgery. AB - BACKGROUND: The establishment of hemostasis without the excessive transfusion of homologous blood and blood products is critical to successful aortic surgery. METHODS AND RESULTS: By using preoperative autologous blood donation and intraoperative blood conservation measures, 85% of patients can undergo aortic surgical procedures without homologous blood or product transfusions, and almost three-quarters of patients will still not have required homologous transfusions by the time of discharge. In contrast, three-quarters of those patients who cannot donate blood preoperatively will require homologous blood transfusions. CONCLUSIONS: The strategy described is safe: our overall survival rate for 204 patients has been 98%, with a 1% incidence of stroke. PMID- 10391353 TI - Anti-fibrinolytic therapy in thoracic aortic surgery. AB - BACKGROUND: Bleeding is an important cause of morbidity in thoracic aortic surgery. METHODS: We reviewed the mechanisms for fibrinolysis in aortic surgery and the propensity for intervention. Several studies have addressed the safety and efficacy of aprotinin. RESULTS: The endothelium regulates the balance between thrombosis and fibrinolysis. During hypothermic circulatory arrest, thrombin generation stimulates protein C production and tissue plasminogen activator release to promote fibrinolysis. Hypothermia also adversely affects platelet function and coagulation. Controversy exists regarding the effectiveness and dangers of antifibrinolytic agents after circulatory arrest. CONCLUSIONS: Fibrinolysis remains problematic during thoracic aortic aneurysm surgery. Heparin management is complicated by aprotinin and insufficient heparin may result in thrombotic events. Aprotinin is safe during rewarming or postoperatively. PMID- 10391354 TI - Management of infected aortic grafts: development of less invasive surgery using cryopreserved homografts. AB - BACKGROUND: Aortic graft infection is associated with significant mortality and morbidity. Total graft replacement with homografts provides an effective treatment. Partial graft replacement at the site of infection may simplify the surgical procedure. METHODS: Between January 1991 and December 1996, homografts were used in 18 patients (mean age, 61+/-12 years; range 41-85) with thoracic (4/18; 22%) or abdominal (14/18; 78%) aortic graft infection. Sepsis was present in 14 patients (78%); 6 (33%) had various aortic fistulae. Total graft replacement using homografts was performed in 14 (78%), and partial graft replacement at the site of infection in 4 patients (22%). RESULTS: Hospital mortality was 11%. During the follow-up period of 22+/-15 months (range, 12-65) there was 1 infection and 1 homograft-related late death after complete homograft replacement, and 1 percutaneous vascular stent placement after partial graft replacement. No other instances of reinfection, suture line rupture or anastomotic aneurysms were observed. CONCLUSION: Total graft replacement with homografts provides an effective treatment for infected aortic grafts. Partial graft replacement at the site of infection is feasible and safe. PMID- 10391355 TI - Management of thoracic aortic graft infections. AB - BACKGROUND: We reviewed our experience managing patients with thoracic aortic graft infections to evaluate their clinical characteristics and outcomes of treatment. METHODS: Records of 20 consecutive patients with thoracic aortic graft infections managed over a 7 year period were retrospectively reviewed. Current follow-up status was obtained for all survivors. RESULTS; Nineteen patients (95%) underwent surgical treatment. Graft excision and in situ replacement were performed using Dacron grafts (10/19, 53%) or cryopreserved homografts (5/19, 26%). Three pseudoaneurysms were managed by debridement and primary repair. Although 30 day postoperative survival was 89% (17/19), in-hospital mortality occurred in 8 patients (42%). Infected thoracoabdominal aortic grafts were universally fatal. Of 6 patients with infected composite valve grafts, both patients who received new composite valve grafts died and all 4 who received homografts survived (p = 0.067). CONCLUSIONS: Infections involving thoracic aortic grafts continue to carry a high mortality rate, especially in patients with polymicrobial infections, thoracoabdominal aortic graft infections, and composite valve graft infections. Use of homografts in the latter situation may improve outcome. PMID- 10391356 TI - A new method for the treatment of graft infection in the thoracic aorta: in situ preservation. AB - BACKGROUND: We have developed a new method to control graft infection by a combination of two procedures, extensive disinfection followed by tissue flap implantation, allowing preservation of the original graft. METHOD: Soon after the diagnosis of graft infection was confirmed, the wound was re-explored, and debridement, irrigation and packing with sponges soaked with 10% iodine solution were employed. This procedure was repeated every 8 hours for the first 48 hours. For the second step, tissue flaps using omentum or muscle were implanted around the graft as well as in dead space, and the wound was closed primarily. MATERIALS: A total of 6 patients were treated: 4 in the acute and 2 in the chronic phase of infection. The original procedures were a Bentall procedure + arch replacement (1), ascending aorta replacement + arch (3) and replacement of the descending aorta (2). In descending aorta cases, an extended thoracoplasty was concomitantly added to eliminate dead space in the pleural cavity. RESULTS: Graft infections were controlled in all 6 patients. One hospital death unrelated to infection was encountered. Five patients were discharged, but 1 died of a stent-graft complication. The follow-up period ranged from 4 months to 10 years. CONCLUSION: Our method of extensive disinfection followed by tissue flap coverage of the graft proved to be highly effective in controlling the serious complication of graft infection associated with surgery of the thoracic aorta. PMID- 10391357 TI - Surgery for acute type A aortic dissection. AB - BACKGROUND: Several innovative approaches have been introduced in the surgical treatment of acute type A aortic dissection. This study examines the effects of these new techniques on the early and late outcomes of patients with this disease. METHODS: The records of patients who had surgery for acute type A aortic dissection during an 18 year interval were reviewed. There were 109 patients: 81 men and 28 women, with a mean age of 57 years, range 23 to 80. Most patients were acutely ill and 15 were in shock at the time of surgery. Operations were performed under cardiopulmonary bypass with femoral artery and right atrial cannulation. In 55 patients, the aorta was clamped and retrograde femoral perfusion was used throughout the procedure (group I). In 54 patients, no clamp was used; under circulatory arrest the primary tear was resected whether in the ascending aorta or transverse arch, and antegrade cardiopulmonary bypass was started after completion of the distal anastomosis (group II). Postoperative computed tomographic or magnetic resonance scans were completed annually. RESULTS: There were 16 operative deaths (15%): 11 (20%) in group I, and 5 (9.2%) in group II (p = 0.10). There were 10 strokes: 8 (14.5%) in group I and 2 (3.7%) in group II (p = 0.05). After a mean follow-up time of 59 +/- 45 months for group I, 31 (56%) patients were alive, and after a mean follow-up time of 45 +/- 26 months for group II, 44 (81%) patients were alive. The actuarial survival of group II was higher than group I, but the difference was not significant (p = 0.09). Postoperatively, a patent false lumen was found in 91% of group I patients and in 59% of group II (p = 0.01). CONCLUSIONS: This study suggests that avoidance of aortic clamping, resection of the primary tear in the ascending aorta or transverse arch, and antegrade perfusion after completion of the distal anastomosis improve the early and late outcomes of surgery for acute type A aortic dissection. PMID- 10391358 TI - Management of descending aortic dissection. AB - BACKGROUND: Experience with 100 consecutive patients with acute dissection of the descending aorta seen at the Yale Center for Thoracic Aortic Disease over a 10 year period is reported. METHODS: Clinical records from the Yale Center for Thoracic Aortic Disease from 1988 to 1998 were analyzed. This computerized data base included information regarding patients' demographics, history, presenting symptomatology, diagnostic imaging, early hospital course, treatment strategy, and long term follow up (office visits, echocardiography, computerized tomography, magnetic resonance imaging, and home phone calls). RESULTS: The average size of the aorta at the time of dissection was 5.05 cm. Nine patients died (six of complications directly related to the thoracic aorta). Sixty of the 91 surviving patients had a benign course, and 31 had a course complicated by rupture (8), vascular occlusion (17), early expansion or extension (12), and continued pain (4); multiple complications were seen in some patients. Forty-two patients came to operation (22 early and 20 late): 32 direct aortic replacements, 6 fenestration procedures, and 4 thromboexclusions. There were six postoperative deaths and six paraplegias. Clinical experience with the alternative procedures of fenestration and thromboexclusion found both procedures safe and effective for selected categories of patients. Review of the literature indicated that direct aortic replacement in the setting of acute descending aortic dissection continues to carry a very high mortality (28%-65%) and paraplegia rate (30%-35%), leaving room for consideration of alternative procedures. CONCLUSIONS: We recommend a "complication-specific" approach to acute descending aortic dissection: medical management with "antiimpulse therapy" for uncomplicated acute descending dissections and surgical intervention for complicated dissections. Surgical therapy varies for the specific complication: for rupture, direct aortic replacement is recommended; for vascular occlusion, fenestration; and for acute expansion or impending rupture, direct aortic replacement, with thromboexclusion as an option. Chronic descending aortic dissection is treated according to general guidelines for descending aortic aneurysms, with operation for symptoms or enlargement > 6.5 cm. PMID- 10391359 TI - Surgery for acute type A aortic dissection: the Hopital Foch experience (1977 1998). AB - BACKGROUND: In 1977, we proposed the use of gelatin-resorcinol-formol (GRF) biological glue during surgery for acute type A aortic dissection. METHODS: From January 1977 to March 1998, 204 patients (146 men and 58 women) aged from 15 to 79 years (mean 54 +/- 11) underwent emergency operation for type A aortic dissection in our institution. One hundred sixty-five patients (84%) were operated on within 48 h after the onset of symptoms. Twenty-eight patients (13.7%) had Marfan's syndrome. In 43 patients (23%), the aortic valve was replaced either independently (6, 3%) or by means of a composite graft (37, 18.1%). Because of the location of the intimal tear, aortic replacement included the transverse arch in 60 patients (29.4%). RESULTS: Hospital mortality was 21% (39 patients): 25% in patients with arch replacement and 19.4% in patients without arch replacement (ns). One hundred sixty-one patients were discharged and followed from 2 months to 21 years postoperatively (mean 85 +/- 66 months). During this interval, 25 patients (15.5%) required reoperation for a total of 33 reoperations. Seven patients (28%) died at reoperation. Upon univariate analysis, presence of Marfan's syndrome (p < 0.05) and absence of arch replacement (p < 0.02) were risk factors for reoperation. Emergency operation (p < 0.01) and thoracoabdominal replacement (p < 0.04) were risk factors for death at reoperation. The actuarial freedom from reoperation (Kaplan-Meier, confidence interval 95%) is 96.1% (90.9%-98.2%) at 1 year, 87.6% (79.8%-92.7%) at 5 years, 80.9% (70.8%-88.1%) at 10 years, and 66.4% (51.1%-78.9%) at 15 years. A total of 39 patients (24.3%) died during follow-up. The presence of Marfan's syndrome (p < 0.01), reoperation (p < 0.02), stroke (p < 0.05), and cardiac failure (p < 0.05) were risk factors for late mortality. The actuarial late survival including hospital mortality is 71.5% (64.3%-77.8%) at 1 year, 66% (58.3%-73%) at 5 years, 56.4% (47.7%-64.7%) at 10 years, and 46.3% (36.4%-56.5%) at 15 years. CONCLUSIONS: The GRF glue has proven extremely useful during emergency initial surgery for acute type A dissection, making the procedure much easier and safer. As a result of this operative improvement, the use of the GRF glue seems to have had a beneficial influence on late results, but these also depend upon the patient's basic condition. PMID- 10391360 TI - Preservation of the aortic valve in acute type A dissection complicated by aortic regurgitation. AB - BACKGROUND: The aim of the present study was to verify the efficacy of preserving the aortic valve in patients with acute type A aortic dissection complicated by significant aortic regurgitation. METHODS: From January 1979 to December 1996, 178 patients (125 males; mean age 57 +/- 9 years) underwent emergency surgery for acute type A aortic dissection, with an overall operative mortality rate of 21%. Based on a retrospective analysis of the preoperative angio- or echocardiographic findings, the 141 survivors were divided into 2 groups: Group 1 (G1) included 80 patients (57%) with no or mild aortic regurgitation, and Group 2 (G2) the remaining 61 patients with moderate-to-severe aortic regurgitation. The native aortic valve was preserved by means of a uniform technique consisting of reconstruction of the aortic root and sinotubular junction in 99 patients (70%) [68 in G1 (85%) and 31 in G2 (51%)]. Forty-two patients required aortic valve (8 patients; 6%) or total root replacement (34 patients; 24%). RESULTS: At a mean follow-up of 4 +/- 3.6 years (range, 6 months to 19 years), 19 of the 99 patients with a preserved aortic valve developed moderate-to-severe aortic insufficiency [19%; 7/68 in G1 (10%) and 12/31 in G2 (39%)]. Multivariate analysis revealed that moderate-to-severe preoperative aortic valve insufficiency was a significant risk factor for development of postoperative aortic valve regurgitation (p = 0.008). Reoperation was necessary in 7 G1 patients (10%) and in 8 G2 patients (26%), with an actuarial freedom from reoperation at 5 and 10 years of 93% +/- 7% and 80% +/- 9% in G1 patients, and 81% +/- 8% and 40% +/- 15% in G2 patients (p = 0.045). CONCLUSIONS: Preservation of the aortic valve and aortic root is recommended in patients with acute type A aortic dissection and absent or mild aortic insufficiency. Patients presenting with moderate-to-severe aortic regurgitation and treated conservatively present an increased risk of recurrent valvular insufficiency. PMID- 10391361 TI - Effects of the bradykinin antagonist Bradycor (deltibant, CP-1027) in severe traumatic brain injury: results of a multi-center, randomized, placebo-controlled trial. American Brain Injury Consortium Study Group. AB - A phase II prospective, randomized, double blind clinical trial of Bradycor, a bradykinin antagonist, was conducted at 31 centers within North America in severely brain injured patients. Patients of Glasgow Coma Score (GCS) 3-8 (n = 139) with at least one reactive pupil were randomized to receive either Bradycor, 3 microg/kg/min or placebo as a continuous intravenous infusion for 5 days, with the infusion beginning within 12 h of the injury. The primary objective was to assess the efficacy of a continuous infusion of Bradycor (3.0 mc/kg/min) in preventing elevation of intracranial pressure (ICP). Other efficacy measures included the effect of Bradycor on the Therapy Intensity Level (TIL), mortality, and functional outcome. A secondary objective was to evaluate the safety of Bradycor in patients with severe brain injury. Randomization was carried out according to a computer generated randomization list. Patients were followed for the first 14 days of hospitalization with long-term outcome assessed at 3 and 6 months after injury. During the infusion and while the ICP monitor was in place, ICP measurements were recorded hourly along with blood pressure and heart rate. A modified version of the TIL was used to record therapeutic interventions hourly, while the ICP was being monitored. Outcome was assessed at 3 and 6 months after injury using the Glasgow Outcome Score (GOS). Bradycor was well tolerated in this patient population, and no adverse events were attributable to the compound. Although positive trends were seen for both ICP and TIL in the Bradycor group, these differences analyzed on a daily basis were not significant. However, a mixed model of variance which included treatment, day, treatment by day interaction, age and GCS revealed that the percentage time ICP of >15 mm Hg on days 4 and 5 was significantly lower in the Bradycor group compared to placebo (p = 0.035). There were fewer deaths in the Bradycor group, which had a 28-day all cause mortality of 20% versus 27% on placebo. Patients treated with Bradycor showed a 10.3% improvement in favorable outcome at 3 months and a 12% improvement in dichotomized GOS at 6 months (p = 0.26). The consistent positive trends seen in ICP, TIL, neuropsychological tests, and, most importantly, 3- and 6-month GOS provide supportive evidence that a bradykinin antagonist may play a neuroprotective role in severe brain injury. PMID- 10391362 TI - Posttreatment with high-dose albumin reduces histopathological damage and improves neurological deficit following fluid percussion brain injury in rats. AB - We have recently shown that high-dose human serum albumin (HSA) therapy confers marked histological protection in experimental middle cerebral artery occlusion. Thus, the purpose of this study was to determine whether treatment with high-dose HSA would protect in a rat model of traumatic brain injury (TBI). Twenty-four hours prior to TBI, the fluid percussion interface was positioned parasagittally over the right cerebral cortex. On the following day, fasted rats were anesthetized with 3% halothane, 70% nitrous oxide, and 30% oxygen and received right parieto-occipital parasagittal fluid-percussion injury (1.5-2.0 atm). Cranial and rectal temperatures were monitored throughout the experiment and held at normothermic levels (36.5-37.5 degrees C) by a warming lamp above the animal's head. The agent (25% human serum albumin, HSA) or vehicle (sodium chloride 0.9%) was administered i.v. (1% of body weight) 15 min after trauma. Behavioral function was evaluated in all rats before and after TBI (at 2 h, 24 h, 48 h, 72 h, and 7 days). Neurological function was graded on a scale of 0-12 (normal score = 0; maximal score = 12). Seven days after TBI, brains were perfusion-fixed, coronal sections at various levels were digitized, and contusion areas in the superficial, middle and deep layers of cortex and in the underlying fimbria were measured. HSA significantly improved the neurological score compared to saline at 24 h, 72 h, and 7 days after TBI (6.0 +/- 0.6 [albumin] versus 8.4 +/- 0.5 [saline]; 3.6 +/- 0.7 versus 6.8 +/- 1.0; and 2.6 +/- 0.6 versus 5.7 +/- 0.8, respectively; p < 0.05). HSA therapy also significantly reduced total contusion area (0.89 +/- 0.2 versus 1.82 +/- 0.3 mm2; p = 0.02). Our findings document that high-concentration albumin therapy instituted 15 min after trauma significantly improves the neurological score and reduces histological damage. We believe that this pharmacological agent may have promising potential for the clinical treatment of brain injury. PMID- 10391363 TI - Severity of experimental brain injury on lactate and free fatty acid accumulation and Evans blue extravasation in the rat cortex and hippocampus. AB - Lactate and free fatty acids (FFAs) were extracted from the cortices and hippocampi of rats subjected to sham operation, or mild (1.25 atm) or moderate (2.0 atm) fluid percussion (FP) injury, and their total tissue concentrations were measured. The elevation of lactate in the injured left cortex (IC) and ipsilateral hippocampus (IH) was significantly greater in the moderate-injury than in the mild-injury group at most test times between 5 min and 48 h after injury. Levels of total FFAs were elevated in the IC and IH to a greater extent and for a longer period after injury in the moderate-injury (up to 48 h) than in the mild-injury group (up to 20 min). In general, the extent and duration of the elevation of most of the individual FFAs (palmitic, stearic, oleic, and arachidonic acids) in the IC and IH were also greater in the moderate-injury group than in the mild-injury group. In the contralateral cortex (CC) and hippocampus (CH), the elevation of lactate and total FFAs (and individual stearic and arachidonic acids) were also greater in the moderate-injury group than in the low-injury group at 5 min after injury. The extravasation of Evans blue in the IC and IH from 3 to 6 h after injury was also the greatest in the moderate-injury group. The hippocampal CA3 neuronal cell loss, but not cortical lesion volume, also increased with the severity of injury. These findings suggest that certain neurochemical, physiological (blood-brain barrier permeability), and morphologic responses increase with the severity of FP brain injury, and such relationships are consistent with the increased behavioral deficits observed with the increase of severity of brain injury. PMID- 10391364 TI - Expression of brain-derived neurotrophic factor, nerve growth factor, and heat shock protein HSP70 following fluid percussion brain injury in rats. AB - Traumatic brain injury can induce the expression of stress-related and neurotrophic genes both within the injury site and in distant regions. These genes may affect severity of damage and/or be neuroprotective. We used in situ hybridization to assess the alterations in expression of the heat shock protein HSP70, nerve growth factor (NGF), and brain-derived neurotrophic factor (BDNF) genes in rat brain following moderate fluid-percussion (F-P) injury at various survival times. HSP70 gene expression was induced at and surrounding the injury site as early as 30 min after trauma. This elevated signal spread ventrally and laterally through the ipsilateral cortex and into the underlying white matter over the next few hours. In addition, there was elevated expression in the temporal hippocampus. BDNF was strongly upregulated in the granular cells of the dentate gyrus and in the CA3 hippocampus 2-6 h after injury. Cortical regions at and near the injury site showed no response at the mRNA level. NGF mRNA increased over the granular cells of the dentate gyrus at early time points. There was also a weaker secondary induction of the NGF gene in the contralateral dentate gyrus of some animals. Cortical response was observed in the entorhinal cortex, bilaterally, but not at the injury site. All three of the studied genes responded quickly to injury, as early as 30 min. The induction of gene expression for neurotrophins in regions remote from areas with histopathology may reflect coupling of gene expression to neuronal excitation, which may be associated with neuroprotection and plasticity. PMID- 10391365 TI - Neuroprotective effect of hypothermia on neuronal injury in diffuse traumatic brain injury coupled with hypoxia and hypotension. AB - It is well established in mechanical head trauma that posttraumatic secondary insults, such as hypoxia and hypotension exacerbate neuronal injury and lead to worse outcome. In this study, the neuroprotective effect of hypothermia on the reduction of supraventricular subcortical neuronal damage was evaluated using an impact-acceleration model of diffuse traumatic brain injury coupled with both moderate and severe periods of hypoxia and hypotension. A total of 135 adult male Sprague-Dawley rats (340-375 g) were divided into three experimental studies: (I) physiological evaluation (n = 36); (II) quantitative analysis of the effect of trauma coupled with moderate and severe hypotension on neuronal damage assessed at 4 (n = 39) and 24 h (n = 24); and (III) the neuroprotective effect of hypothermia following moderate secondary insult (n = 36). Induction of hypothermia occurred at 15 min postinjury, to a level of 30 degrees C for 60 min. At the designated time points (4 and 24 h), the animals were sacrificed via standard transcardial perfusion techniques for histological processing. Quantitative assessment of neuronal damage using routine H&E staining at 4 hours showed neuronal damage which correlated with the severity of secondary insult. Animals exposed to trauma alone had a mean number of damaged neurons of 7.61 +/- 3.08/high powered field (hpf) compared with a mean of 1.21 +/- 0.30/hpf in the sham operated group (p = 0.015). Animals exposed to trauma with 10 min of hypoxia and hypotension (THH-10) showed a statistically significant number of damaged neurons compared to the sham-operated animals (7.50 +/- 2.15 damaged neurons/hpf, p = 0.013), whereas, neuronal damage in animals undergoing trauma with a 30-min secondary insult of hypoxia and hypotension (THH-30) was markedly increased (100 +/- 30.20/hpf, p = 0.002). Statistical analysis showed no significant difference in neuronal damage in animals subjected to secondary insult alone. At 24 h, the evolution of neuronal damage in the trauma alone group (5.08 +/- 1.63/hpf) was relatively static; however, there was a remarkable increase in the neuronal damage of the THH-10 group (29.88 50 +/- 8.20/hpf). However, hypothermia provided nearly complete protection against secondary insults, and neuronal damage was equal to that of the trauma alone group (p = 0.42). The results of this study confirm that hypothermia provides remarkable protection against the adverse effects of neuronal damage exacerbated by secondary injury. This study also presents a new model of secondary insult, which can be used experimentally to further define the mechanism of increased vulnerability of the injured brain. PMID- 10391366 TI - Alterations in BDNF and trkB mRNA levels in the cerebral cortex following experimental brain trauma in rats. AB - Recent studies have suggested that brain-derived neurotrophic factor (BNDF) and its receptor, trkB, may provide neuroprotection following injury to the central nervous system. Conversely, other studies have implicated BDNF as a contributing factor to neurodegenerative events that occur following injury. In order to further investigate the role of BDNF in neuroprotection, we subjected adult rats to a lateral fluid percussion (FP) injury of moderate severity (2.0-2.1 atm) or sham injury. After survival periods of 1, 3, 6, 24, or 72 h, the brains were processed for the in situ hybridization localization of BDNF and trkB mRNAs using 35S-labeled cRNA probes. Hybridization levels were compared between injured and sham animals for regions of the cortex that were located within, adjacent to, and remote from the site of the cortical contusion. BDNF mRNA levels were significantly decreased in the injured cortex at 72 h, increased in adjacent cortical areas at 3 h, and increased bilaterally in the piriform cortex from 3 to 24 h post-FP injury. Expression of trkB mRNA was significantly decreased at all postinjury time-points in the injured cortex and at 24 h in the adjacent cortex. These results demonstrate that, following lateral FP injury, BDNF and trkB mRNA levels are decreased in cortical regions that contain degenerating neurons, generally unchanged in adjacent regions, and increased in remote areas. Thus, injury-induced decreases in the expression of BDNF and trkB may confer vulnerability to neurons within the cortical contusion. PMID- 10391367 TI - Postinjury cyclosporin A administration limits axonal damage and disconnection in traumatic brain injury. AB - Recent observations concerning presumed calcium-induced mitochondrial damage and focal intraaxonal proteolysis in the pathogenesis of traumatic axonal injury (TAI) have opened new perspectives for therapeutic intervention. Studies from our laboratory demonstrated that cyclosporin A (CsA), a potent inhibitor of Ca2+ induced mitochondrial damage, administered 30 min prior to traumatic brain injury preserved mitochondrial integrity in those axonal foci destined to undergo delayed disconnection. We attributed this neuroprotection to the inhibition by CsA of mitochondrial permeability transition (MPT). Additional experiments proved that CsA pretreatment also significantly reduced calcium-induced, calpain mediated spectrin proteolysis (CMSP) and neurofilament compaction (NFC), pivotal events in the pathogenesis of axonal failure and disconnection. Given these provocative findings the goal of the current study was to evaluate the potential of CsA to inhibit calcium-induced axonal damage in a more clinically relevant postinjury treatment paradigm. To this end, cyclosporin A was administered intrathecally to Sprague Dawley rats 30 min following impact acceleration traumatic brain injury. The first group of animals were sacrificed 120 min postinjury and the density of CMSP and NFC immunoreactive damaged axonal segments of CsA-treated and vehicle-treated injured animals were quantitatively analyzed. A second group of CsA- versus vehicle-treated rats was sacrificed at 24 h postinjury to compare the density of damaged axons displaying beta amyloid precursor protein (APP) immunoreactivity, a signature protein of axonal perturbation and disconnection. Postinjury CsA administration resulted in a significant decrease (>60%) in CMSP/NFC immunoreactivity in corticospinal tracts and medial longitudinal fasciculi. A similar decrease was detected in the density of APP immunoreactive damaged axons, indicating an attenuation of axonal disconnection at 24 h postinjury in CsA-treated animals. These results once again suggest that the maintenance of the functional integrity of the mitochondria can prevent TAI, presumably via the preservation of the local energy homeostasis of the axon. Moreover and perhaps more importantly, these studies also demonstrate the efficacy of CsA administration when given in the early posttraumatic period. Collectively, our findings suggest that a therapeutic window exists for the use of drugs targeting mitochondria and energy regulation in traumatic brain injury. PMID- 10391368 TI - Human spinal cord retains substantial structural mass in chronic stages after injury. AB - In chronic stages of human spinal cord injury, atrophy of the cord has been reported in regions both at and distant to the injury site. Local cord atrophy results from the direct effects of bony impact and ischemia, whereas distant atrophy results from anterograde (Wallerian) and retrograde axonal degeneration. However, the actual extent of degenerative changes in the chronically injured human spinal cord both at and remote from the injury site has rarely been reported, and has not been rigorously quantified to date. In the present study, we quantified the extent of spinal cord atrophy in 12 humans with chronic injury (2-34 years posttrauma) utilizing quantitative stereological assessment of spinal cord magnetic resonance images, and compared the results to uninjured human spinal cords. Focal cystic atrophy of the cord, characterized by signal attenuation on T1-weighted images, was regularly present at the actual site of impact injury and replaced a mean longitudinal area equaling less than one spinal cord segment in length (2.01 +/- 0.60 cm2, or a loss of 89.3 +/- 17.4% of the longitudinal area of one spinal cord segment). Spinal cord segments immediately rostral to the zone of cystic degeneration showed atrophy of only 19.4 +/- 7.5% of normal cord longitudinal area, and spinal cord segments immediately caudal to the zone of cystic degeneration showed atrophy of 16.5 +/- 4.1% of normal cord longitudinal area. Extensive spinal cord atrophy extending beyond the region of injury occurred in two of twelve cases (16.7%), and both were caused by late syrinx formation. Thus, spinal cord atrophy after trauma remains primarily restricted to the original site of injury. Experimental neural repair strategies should take into account the importance of "bridging" relatively short zones of cystic atrophy, then promoting axonal regeneration through potentially long segments of remaining cord parenchyma. PMID- 10391369 TI - Changes in neuronal receptive field characteristics in caudal brain stem following chronic spinal cord injury. AB - Chronic spinal cord injury pain is poorly understood and, thus, not effectively relieved by traditional treatments. In the present study, a variety of partial, severe and sham chronic spinal lesions were made in 31 male rats at spinal level T8. During routine care/handling and brief behavioral testing of the animals throughout the 30-day recovery period, the majority of those with severe contusion injuries (verified histologically) showed signs of mechanical hypersensitivity on the dorsolateral trunk just rostral to the level of injury (i.e., upper thoracic territory). Terminal electrophysiological experiments were performed on all rats (urethane anesthesia). Single unit recordings were made at two supraspinal locations within the caudal brainstem, the nucleus reticularis gigantocellularis and nucleus reticularis gigantocellularis pars alpha. Neurons in these areas normally receive bilateral nociceptive somatovisceral inputs from many parts of the body. Seventy-three percent of the animals with severe contusion injuries developed novel low-threshold neuronal responses to stimulation of the dorsolateral trunk (upper thoracic territory). This amount was significantly greater than for animals with more moderate spinal lesions (dorsal or lateral hemisection; 29% and 25%, respectively) or sham controls (0%). These data suggest (1) that the spinal contusion is a reliable model for studies of the neural mechanisms that underly central spinal cord injury-related pain and (2) that the caudal brainstem is one supraspinal location where neurons undergo significant changes in responsiveness following severe chronic spinal cord injury. The observed plasticity is likely part of the central reorganization producing the multitude of sensory disturbances that surface following spinal cord injury. PMID- 10391370 TI - Alpha-melanocyte stimulating hormone promotes regrowth of injured axons in the adult rat spinal cord. AB - Peptides related to melanotropin (alphaMSH) and corticotropin (ACTH), collectively termed melanocortins, are known to improve the postlesion repair of injured peripheral nerves. In addition, melanocortins exert trophic effects on the outgrowth of neurites from central nervous system neurons in vitro. Here we report, for the first time, the stimulation by alpha-MSH of spinal neurite outgrowth in vivo after injury. In the in vivo model, spinal cord trauma was produced at lower thoracic spinal levels of adult rats. Under a surgical microscope a laminectomy was performed exposing the dorsum of the spinal cord. Then the dura was cut longitudinally and the dorsal columns were identified. Iridectomy scissors were used to transect the dorsal half of the spinal cord bilaterally, thereby completely lesioning the main corticospinal tract component. Then the lesion gap was immediately filled with a solid collagen matrix. Ingrowth of fibers was quantified using an advanced image analyser using a video image of sections transmitted by a camera. In the control situation virtually no ingrowth of sprouting injured fibers into the collagen implant in the lesion gap was seen. However, when the collagen matrix contained 10(-8) M alpha-MSH, a profound and significant stimulation of fiber ingrowth into the implant was observed (alpha MSH, 21.5 +/- 2.9%; control, 1.4 +/- 0.6% p < 0.01). A small percentage of these ingrowing fibers was CGRP-immunoreactive (17.0 +/- 4%), whereas no serotonergic ingrowth was observed. Furthermore, we found that local application of alpha-MSH directs a substantial amount of lesioned anterogradely labelled corticospinal tract axons to regrow into the collagen implant (alpha-MSH, 15.2 +/- 5.2%; control, 0.5 +/- 0.3%, p < 0.01). The observed fiber ingrowth is not accompanied by an invasion of astroglial or reactive microglial cells into the implant. In conclusion, inclusion of alpha-MSH in the collagen implant stimulates the regrowth of injured axons in the adult rat spinal cord. PMID- 10391371 TI - 1997 ESACP consensus report on diagnostic DNA image cytometry. Part I: basic considerations and recommendations for preparation, measurement and interpretation. European Society for Analytical Cellular Pathology. AB - A task force of invited experts in the field of diagnostic DNA image cytometry, especially consisting of participants from the PRESS (Prototype Reference Standard Slides) and EUROPATH (European Pathology Assisted by Telematics for Healthcare) projects, but open to any other scientist or physician revealing experience in that new diagnostic procedure (names are given in the Annex A) agreed upon the following updated consensus report during the 5th International Congress of the ESACP 1997 in Oslo. This report is based on the preceeding one [9] and on results of the above mentioned European research projects. It deals with the following items: Biological background and aims of DNA image cytometry, Principles of the method, Basic performance standards, Diagnostic interpretation of DNA measurements, Recommendations for practical use. PMID- 10391372 TI - 1997 ESACP consensus report on diagnostic DNA image cytometry. Part II: Specific recommendations for quality assurance. European Society for Analytical Cellular Pathology. PMID- 10391373 TI - Chromatin texture from hematoxylin stained thyroid lesions. AB - Quantitative aspects of cytology and histology should be considered in diagnostic standardisation processes. The present paper summarises the cytological differences detected in 75 thyroid lesions using a computerized textural analysis. Cells stained with progressive hematoxylin and taken from paraffin blocks were overlaid with the extracted texture. This technique was based on the lineal detection of a grey level gradient of the common logarithm of the integrated optical density (IOD) of each nucleus. Diffuse and nodular goiters (36 cases) were demonstrated to be composed of small cells containing high density texture that, on microscopical visual inspection, gave a "salt and pepper" appearance. The adenomatous goiters (2 cases) and adenomas (26 cases) were composed of low texture cells with a visual "blurry or smudgy" chromatin, while the atypical adenomas with capsular invasion (4 cases) were characterised by a "woodworm" nuclear appearance that produced the highest texture of the series. Finally, encapsulated folliculo-papillary carcinomas (3 cases) were composed of large clear nuclei with high IOD, low texture, and scattered lines that resulted in an "empty grape skin" aspect. Our findings seam to confirm the suitability of computerized textural techniques that aid in recognizing cell microscopic features objectively. The one used in the present work, based on a mathematical function of the DNA content of each individual nucleus (IOD), fulfills all microscopy detection criteria. PMID- 10391374 TI - Morphometric measurements to quantify the cerulein induced hyperstimulatory pancreatitis of rats under the protective effect of lectins. AB - In preceding papers we demonstrated an inhibitory effect of wheat germ agglutinin (WGA) and Ulex europaeus agglutinin (UEA) on the cholecystokinin (CCK) binding to the CCK receptor of rat pancreatic cells and also on the CCK induced Ca2+ release and alpha-amylase secretion in vitro as well as on pancreatic secretion of intact rats in vivo. In the present study we show the same inhibitory effect of both lectins on the cerulein pancreatitis of rats. This acute pancreatitis was induced by supramaximal injections (5 microg/kg/h i.v. or 10 microg/kg/h i.p.) of the CCK analogue cerulein in rats every hour. To monitor the degree of pancreatitis, we measured the number and diameter of injury vacuoles in the pancreatic acinar cells as one of the most important signs of this type of pancreatitis by light microscopic morphometry with two different systems on paraffin sections. Furthermore, the serum alpha-amylase activity was measured biochemically. We found a correlation between the diameter of vacuoles inside the acinar cells and the serum enzyme activity up to 24 h. The simultaneous i.p. administration of cerulein and WGA or UEA in a dosage of 125 microg/kg/h for 8 h led to a reduction of vacuolar diameter from 13.1+/-2.0 microm (cerulein) to 7.5+/-1.1 microm (cerulein + WGA) or 7.2+/-1.3 microm (cerulein + UEA). The serum amylase activity was reduced from 63.7+/-15.8 mmol/l x min (cerulein) to 37.7+/-11.8 (cerulein + WGA) or 39.4; +52.9; -31.1 (cerulein + UEA-I). Both parameters allow the grading this special type of pancreatitis to demonstrate the protective effect of the lectins. PMID- 10391376 TI - Automatic cell segmentation in cyto- and histometry using dominant contour feature points. AB - Automatic cell segmentation has various application potentials in cytometry and histometry. In this paper, an automatic cluster (touching) cell segmentation approach using the dominant contour feature points has been presented. Dominant feature points are the locations of indentation on the contour of the cluster. First, dominant feature points on the contour of the cluster are detected by distance profile. Next, using shape features of the cells, these feature points are selected for segmentation. We compared the results of the proposed method with manual segmentation and observed that the method has an overall accuracy about to 82%. PMID- 10391375 TI - Relationship between AgNOR proteins, Ki-67 antigen, p53 immunophenotype and differentiation markers in archival breast carcinomas. AB - The present study investigated (i) the relationship between standardised morphometric AgNOR parameters (argyrophilic nucleolar organiser region-associated proteins) and MIB1 growth fraction, and (ii) their correlation with immunohistochemical p53, sex steroid receptor status and histopathological differentiation grade in serial paraffin sections from 39 breast carcinomas. Ten sections were double-stained for AgNOR/MIB1. AgNOR parameters correlated significantly with MIB1 growth fraction and p53 protein expression. Significant inverse correlation was found between proliferation markers and oestrogen/progesterone receptor status and histopathological grade. AgNOR expression was significantly higher in cycling (MIB1 positive) tumour cells, than in resting (MIB1 negative) ones, however with exceptions. We conclude, that standardised AgNOR parameters correlate with markers of increased malignant potential in breast carcinomas. However, AgNORs seem to reflect proliferation independent cellular and nucleolar activity of tumour cells, as well. We recommend the use of standardised AgNOR analysis for obtaining sound results in routine paraffin sections. PMID- 10391377 TI - Acquisition of lipoproteins in the procyclic form of Trypanosoma brucei. AB - The procyclic form of Trypanosoma brucei binds and internalizes bovine high density lipoprotein (HDL) particles in a saturable process; the binding and uptake of (125)I-labeled HDL are inhibited by excess unlabeled HDL. We calculated that each procyclic trypanosome exposes approximately 1.0 x 10(6) binding sites for bovine HDL, with an equilibrium dissociation constant (Kd) of approximately 1.26 x 10(-7) M. Uptake of HDL particles does not occur at 4 degrees C. At 28 degrees C, a significant amount of the internalized HDL particles were efficiently degraded through a process that is sensitive to the presence of 50 microM chloroquine. These results suggested that the uptake of HDL particles in procyclic T. brucei may occur via receptor mediated endocytosis, leading to proteolytic degradation of the particles in an acidic and endocytic compartment. PMID- 10391378 TI - Differential tissue distribution of diverse clones of Trypanosoma cruzi in infected mice. AB - Chagas disease, caused by the protozoan Trypanosoma cruzi, presents variable clinical course but the phenomena underlying this variability remain largely unknown. T. cruzi has a clonal population structure and infecting strains are often multiclonal. T. cruzi genetic variability could be a determinant of differential tissue tropism or distribution and consequently of the clinical forms of the disease. We tested this hypothesis by using low-stringency single specific primer polymerase chain reaction (LSSP-PCR) to type genetically the parasites in tissues of experimental infected mice. BALB/c mice were simultaneously inoculated with two different T. cruzi populations (JG strain and Coll.7G2 clone). Doubly infected animals showed clear differential tissue distribution for the two populations (chronic phase). Our results indicate a significant influence of the genetic polymorphism of infecting T. cruzi populations in the pathogenesis of chronic Chagas disease. PMID- 10391379 TI - Organization of telomeric and sub-telomeric regions of chromosomes from the protozoan parasite Trypanosoma cruzi. AB - We present here a characterization of the telomeric and subtelomeric regions of Trypanosoma cruzi chromosomes, using three types of recombinants: cosmids from a genomic library, clones obtained by a vector-adaptor protocol, and a recombinant fragment cloned by a Bal31 trimming protocol. The last nine nucleotides of the T. cruzi overhang are 5'-GGGTTAGGG-3', and there are from 9 to 50 copies of the hexameric repeat 5'-TTAGGG-3', followed by a 189-bp junction sequence common to all recombinants. The subtelomeric region is made of sequences associated with the gp85/sialidase gene family, and/or sequences derived from SIRE, a retrotransposon-like sequence, and also the retrotransposon L1Tc. We discuss the possible implications of this genome organization. PMID- 10391380 TI - Structural characterisation and pharmacology of KHEYLRFamide (AF2) and KSAYMRFamide (PF3/AF8) from Haemonchus contortus. AB - The FMRFamide-related peptides (FaRPs), KHEYLRFamide (AF2) and KSAYMRFamide (PF3) were structurally characterised from the parasitic nematode of sheep, Haemonchus contortus (MH isolate). Both peptides were sequenced in a single gas-phase sequencing run and their structure confirmed by mass spectrometry which identified peptides of 920 Da (C-terminally amidated AF2) and 902/918 Da (C terminally amidated non-oxidised/oxidised PF3, respectively). AF2 had inhibitory effects on H. contortus muscle and inhibited acetylcholine (ACh, 10 microM) induced contractions, with a threshold for activity of 1 microM. PF3 induced concentration-dependent contractions of H. contortus (activity threshold, 10 nM) and enhanced ACh contractions. Compared with the MH isolate, an isolate of H. contortus which has reduced sensitivity to cholinergic drugs (Lawes isolate) was less sensitive to the effects of PF3. The concentration-response curves for the cholinergic compounds ACh and levamisole (LEV), and PF3, but not a control, KPNFIRFamide (PF4), showed a statistically similar shift. This study implicates PF3 in the modulation of cholinergic function in H. contortus. PMID- 10391381 TI - Plasmodium falciparum-infected erythrocyte adhesion to the type 3 repeat domain of thrombospondin-1 is mediated by a modified band 3 protein. AB - Previously, the binding site for the Plasmodium falciparum-infected erythrocyte (PE) was determined to be the C-terminal 120 or 140 kDa region but not the N terminal 25 kDa domain of thrombospondin (TSP). In this work, we have localized the TSP binding site for PE more precisely. PE adhered to glutathione-S transferase-fusion proteins containing the type 3 repeat (T3) of TSP, but not to other functional domains of TSP (i.e. N-terminal domain, procollagen domain, type 1 and 2 repeat, and C-terminal domain). Soluble T3 inhibited PE binding to immobilized TSP. PE binding to immobilized T3 was inhibited by soluble TSP, a monoclonal antibody directed against the T3, glycine-arginine-glycine-aspartic acid-serine-proline (GRGDSP) peptide, and *cysteine-GRGDSP-cysteine*, where *cysteine and cysteine* form a disulfide linkage, suggesting involvement of an RGD-containing motif in the T3. In support of this, a fusion protein which excluded the RGD motif showed no PE binding activity. Earlier it was shown that the amino acid sequence of the band 3 protein, histidine-proline-leucine glutamine-lysine-threonine-tyrosine (HPLQKTY), was exposed on PE and mediated PE binding to TSP. Monoclonal antibodies, which recognize HPLQKTY and inhibit PE binding to TSP, also inhibited PE binding to the T3. The involvement of the sequence was confirmed by the fact that an octamer of HPLQKTY-containing peptide bound to the T3 but not to the RGD motif-excluded fusion protein and the binding to T3 was inhibited by GRGDSP peptide. Thus, PE binding to the T3 domain of TSP is mediated by the peptidic sequence HPLQKTY of band 3 which is exposed on PE. PMID- 10391382 TI - Typing of Leishmania lipophosphoglycans by electrospray mass spectrometry. AB - A method has been developed to identify the repeating phosphosaccharide units of Leishmania lipophosphoglycans using electrospray mass-spectrometry (ES-MS). Cone voltage-induced fragmentation of intact lipophosphoglycan was found to be as effective as analysis of mild acid hydrolysates in identifying the degree of modification of the repeating units of lipophosphoglycans derived from Leishmania mexicana and Leishmania major. This finding was exploited in a 'rapid-analysis' method in which a crude organic extract of approximately 2 x 10(9) L. major promastigote cells was loaded onto a reverse-phase cartridge for immediate elution into the mass-spectrometer. Using this approach, it was possible to identify the repeating units by total ion scanning and scanning for parents of the m/z 79 (PO3-) fragment ion. This approach is suitable for quick-typing of lipophosphoglycan repeats and was shown to detect alterations in repeat side chains caused by: (1) culturing L. major promastigotes in the presence of L fucose; and (2) in vitro metacyclogenesis of L. major promastigotes. It is anticipated that the method will be applicable to small samples of cultured field isolates or genetically-manipulated strains. PMID- 10391383 TI - Gene organization of rab6, a marker for the novel Golgi of Plasmodium. PMID- 10391384 TI - Replacement of Leishmania N-acetylglucosamine-1-phosphate transferase gene requires episomal rescue. PMID- 10391385 TI - A chromosome-specific dispersed gene family in Trypanosoma cruzi. PMID- 10391386 TI - Inactivation of the gene encoding the flagellar pocket protein, CRAM, in African trypanosomes. PMID- 10391388 TI - The production of the osmiophilic body protein Pfg377 is associated with stage of maturation and sex in Plasmodium falciparum gametocytes. PMID- 10391387 TI - A Plasmodium falciparum apical membrane antigen-1 (AMA-1) gene apparently generated by intragenic recombination. PMID- 10391389 TI - Small is beautiful - but too cheap. PMID- 10391390 TI - Diabetes epidemiology: present and future. PMID- 10391391 TI - Angiotensin-converting enzyme gene and diabetes mellitus. AB - AIMS: The association of the insertion/deletion polymorphism in the angiotensin converting enzyme (ACE) gene with cardiovascular disease and diabetic nephropathy remains a controversial issue. This review aims to give an overview of the research to date assessing the impact of the ACE polymorphism in Type 1 and Type 2 diabetes mellitus (DM). METHODS: A systematic review of the literature was performed in the databases of MEDLINE, PubMed and EMBASE for the key words 'diabetes mellitus', 'diabetic nephropathy', 'ACE polymorphism' and 'genotype' and relevant articles were considered. RESULTS: A meta-analysis assessing the influence of the ACE polymorphism on disease susceptibility demonstrated significant odds ratios in individuals with the DD genotype for coronary heart disease, myocardial infarction and both diabetic and nondiabetic renal disease. No association was found for left ventricular hypertrophy or hypertension in nondiabetic subjects. CONCLUSIONS: The ACE polymorphism appears to have a significant impact on the progression of diabetic nephropathy and may have therapeutic implications for identifying those individuals resistant to the effects of ACE inhibitors. It also appears to be indicative of an increased vascular risk in diabetic patients; however, larger prospective studies are required to clarify this situation. PMID- 10391392 TI - The British Diabetic Association Cohort Study, I: all-cause mortality in patients with insulin-treated diabetes mellitus. AB - AIMS: To assess mortality in patients with diabetes incident under the age of 30 years. METHODS: A cohort of 23 752 diabetic patients diagnosed under the age of 30 years from throughout the United Kingdom was identified during 1972-93 and followed up to February 1997. Following notification of deaths during this period, age- and sex-specific mortality rates, attributable risks and standardized mortality rates were calculated. RESULTS: The 23 752 patients contributed a total of 317 522 person-years of follow-up, an average of 13.4 years per subject. During follow-up 949 deaths occurred in patients between the ages of 1 and 84 years, 566 in males and 383 in females. All-cause mortality rates in the patients with diabetes exceeded those in the general population at all ages and within the cohort were higher for males than females at all ages except between 5 and 15 years. The relative risk of death (standardized mortality ratio, SMR), was higher for females than males at all ages, being 4.0 (95% CI 3.6 4.4) for females and 2.7 (2.5-2.9) for males overall, but reaching a peak of 5.7 (4.7-7.0) in females aged 20-29, and of 4.0 (3.1-5.0) in males aged 40-49. Attributable risks, or the excess deaths in persons with diabetes compared with the general population, increased with age in both sexes. CONCLUSIONS: This is the first study from the UK of young patients diagnosed with diabetes that is large enough to calculate detailed age-specific mortality rates. This study provides a baseline for further studies of mortality and change in mortality within the United Kingdom. PMID- 10391393 TI - The British Diabetic Association Cohort Study, II: cause-specific mortality in patients with insulin-treated diabetes mellitus. AB - AIMS: To measure cause-specific mortality, by age, in patients with insulin treated diabetes incident at a young age. METHODS: A cohort of 23 752 patients with insulin-treated diabetes diagnosed under the age of 30 years, from throughout the United Kingdom, was identified during 1972-93 and followed to February 1997. Death certificates have been obtained for deaths during the follow up period and cause-specific mortality rates and standardized mortality ratios by age and sex are reported. RESULTS: During the follow-up period 949 deaths occurred and at all ages mortality rates were considerably higher than in the general population. Acute metabolic complications of diabetes were the greatest single cause of excess death under the age of 30 years. Cardiovascular disease was responsible for the greatest proportion of the deaths from the age of 30 years onwards. CONCLUSIONS: Deaths in patients with diabetes diagnosed under the age of 30 have been reported and comparisons drawn with mortality in the general population. To reduce these deaths attention must be paid both to the prevention of acute metabolic deaths and the early detection and treatment of cardiovascular disease and associated risk factors. PMID- 10391394 TI - A population-based survey of frequencies of self-reported spontaneous and induced abortions in Danish women with Type 1 diabetes mellitus. Danish IDDM Epidemiology and Genetics Group. AB - AIMS: Whether pregnant women with Type 1 diabetes mellitus (Type 1 DM) have an increased risk of abortions is controversial. The aim of the present Danish population-based study of 33% of the Danish population was to describe the pattern of self-reported miscarriage and stillbirths from 1304 women with Type 1 DM. METHODS: Data were obtained by questionnaire. The current age of the women was 20-65 years and their age at diabetes onset was 30 years or less. RESULTS: The frequency of spontaneous abortions (SA) and induced abortions (IA) reported from women diagnosed with Type 1 DM prior to pregnancy was 17.5% (95% CI 15.5 19.9%) and 18.0%. (95% CI 16.0-20.0%), respectively. No significant differences in abortion frequencies before or after 1980 were reported. Previous findings of a decreasing stillbirth-rate in diabetic pregnancies during the last decades were supported. CONCLUSIONS: The reported SA frequency of 17.5% (95% CI 15.5-19.9%) in pregnant women with Type 1 DM is higher than previously reported SA rates of 10 12% in Danish nondiabetic women and the SA rate is more than twice the SA rate found in a previous Danish study from a highly specialized obstetrical centre for diabetic women. These data suggest an urgent need for further improvement in the general management of Danish pregnant women with Type 1 DM.